PMID- 15521645 TI - Cytogenetic characterization of the Zebra Mussel Dreissena polymorpha (Pallas) from Miedwie Lake, Poland. AB - Cytogenetic characterization of D. polymorpha was carried out using banding techniques such as C-banding, fluorochrome CMA3 and silver nitrate treatment. The diploid chromosome number of both investigated D. polymorpha forms (typical and albinotic) was the same 2n = 32 (NF = 56). The karyotype consisted of 5 pairs of metacentric, 7 pairs of submetacentric and four pairs of subtelo-acrocentric chromosomes. Ag-NORs were located in the telomeric position on the largest subtelo-acrocentric chromosome pair. C banding patterns indicate many sites of constitutive heterochromatin mainly located in the telomeric regions and interstitially in some chromosomes. CMA3-sites were observed in almost all chromosomes; apart from the Ag-NORs sites, they were located terminally on the chromosome arms and interstitially on three chromosome pairs. Sixteen chromosomes could be counted at the diakinesis stage of meiosis. No differences in banding chromosome patterns were found neither between both analyzed forms of D. polymorpha nor between males and females. PMID- 15521646 TI - Experimental hybridisation between X0 and XY chromosome races in the grasshopper Podisma sapporensis Shir. (Orthoptera, Acrididae). I. Cytological analysis of embryos and F1 hybrids. AB - The results of experimental hybridisation between some chromosome subraces belonging to the X0 and XY chromosome races of the brachypterous grasshopper P. sapporensis are presented. Pre-zygotic reproductive isolation mechanisms in experimental pairs were not confirmed. In crossings of XY-standard x X0-standard and XY-standard x X0-Naganuma chromosome subraces, a zygotic barrier has been found. All embryos of XY-standard x X0-standard crosses and the vast majority of embryos of XY-standard x X0-Naganuma crosses were obtained from female diploid or haploid/diploid cells as a result of parthenogenesis. In very rare cases, when the zygotic barriers had been surmounted, normal embryo heterozygotes and a F1 hybrid generation were obtained in XY-standard x X0-Naganuma crosses. On the contrary, crosses between the XY-Tanno and X0-standard subraces gave viable offspring in spite of many chromosome differences such as a X-A translocation and fixed pericentric inversions in four pairs of autosomes. The results obtained do not support the hypothesis that chromosomal differences play a key role in restricting gene flow between X0 and XY races of P. sapporensis. The presence of crossing barriers explains the phenomena of the purity of the X0 and XY chromosomes races. PMID- 15521647 TI - Chromosome banding studies by replication and restriction enzyme treatment in vendace (Coregonus albula) (Salmonidae, Salmoniformes). AB - Chromosome banding studies were performed in vendace, Coregonus albula. Original data on distribution of early and late replication regions, restriction sites (AluI, DdeI, HinfI and HaeIII) on chromosomes in this coregonid fish have been used to analyse karyotype heterochromatin differentiation. Heterochromatic bands (C-positive and not digested by restriction enzymes) have been identified as late replicating regions. Extra bands produced by the applied methods have permitted the identification of several homologous pairs. The centromeres were differentially digested by the restriction enzymes. The studied population seems to be homogenic regarding karyotype characteristics. PMID- 15521648 TI - Cytogenetic variation in farmed stock of Dybowski's sika deer. AB - The chromosome constitution of Dybowski's sika deer was studied on the basis of 15 samples obtained from farmed stock maintained in an enclosure. The diploid chromosome number was 2n=68, 2n=67 and 2n=66. The constitutive heterochromatin (C bands) was located in the centromeric regions of all acrocentric chromosomes. Metacentric chromosomes were C-negative. Chromosomes of three pairs proved to be NORs carriers. The size polymorphism of silver deposits was identified in two animals. A cytogenetic analysis indicated that the farmed stock of Dybowski's sika deer demonstrates considerable variation. The chromosome polymorphism observed may be a valuable marker for the management and preservation of this species. PMID- 15521649 TI - Karyotype of the cyprinid fish Alburnoides bipunctatus (Cyprinidae) from the Tigris River. AB - Chromosome numbers and karyotypes of Cyprinid fish Alburnoides bipunctatus (Bloch, 1,782) from the River Tigris were determined by the chromosome preparation technique from uncultured kidney cells. The diploid chromosome number 2n=50, was composed of 8 pairs of metacentric, 11 pairs of submetacentric and 6 pairs of subtelo-acrocentric chromosomes (NF=88). Sex chromosomes were not determined in the this species. The results were briefly discussed with other, previously conducted studies. PMID- 15521650 TI - Notes on chromosome numbers and C-banding patterns in karyotypes of some weevils from central Europe (Coleoptera, Curculionoidea: Apionidae, Nanophyidae, Curculionidae). AB - Chromosome numbers and C-banding patterns of sixteen weevil species are presented. The obtained results confirm the existence of two groups of species with either a small or large amount of heterochromatin in the karyotype. The first group comprises twelve species (Apionidae: Oxystoma cerdo, Eutrichapion melancholicum, Ceratapion penetrans, Ceratapion austriacum, Squamapion flavimanum, Rhopalapion longirostre; Nanophyidae: Nanophyes marmoratus; Curculionidae: Centricnemus (=Peritelus) leucogrammus, Sitona humeralis, Sitona lineatus, Sitona macularis, Sitona suturalis). In weevils with a small amount of heterochromatin, tiny grains on the nucleus during interphase are visible, afterwards appearing as dark dots during mitotic and meiotic prophase. The second group comprises four species from the curculionid subfamily Cryptorhynchinae (Acalles camelus, Acalles commutatus, Acalles echinatus, Ruteria hypocrita) which possess much larger heteropycnotic chromosome parts visible during all nuclear divisions. The species examined have pericentromeric C-bands on autosomes and on the X chromosome. PMID- 15521651 TI - Cytogenetic studies of Glomeris (Diplopoda: Glomeridae). AB - Karyotypes and meiosis of Glomeris hexasticha and G. connexa (Diplopoda: Glomeridae) from Poland were described using C-heterochromatin distribution and observations of the location of NORs. These species were characterized by 2n>=16 and the XY sex determination system. Differences were found in the amount of C heterochromatin in X and Y chromosomes between the studied species. In G. hexasticha, supernumerary B chromosomes were described. PMID- 15521652 TI - Adrenomedullary and glycemic responses to ACTH, corticosterone, aldosterone, epinephrine and norepinephrine administrations in the soft-shelled turtle. AB - The aim of the current investigation was to ascertain the role of ACTH and adrenal hormones on adrenomedullary and glycemic functions in soft-shelled turtles, Lissemys punctata punctata. All the experiments were carried out on sexually immature animals. Findings revealed that: (1) ACTH administration (0.5 IU/1.0 IU/2.0 IU per 100 g body wt. daily for 10 days) in all doses stimulated adrenomedullary function by increasing medullary cell nuclear diameter with elevations of norepinephrine, epinephrine and blood sugar levels. Only moderate and higher doses (50 microg/100 microg per 100 g body wt. daily for 10 days) of dexamethasone suppressed adrenomedullary activity and blood sugar level by reversing the changes to those of ACTH; the responses were dose-dependent. But these changes were no longer observed after ACTH treatment in dexamethasone (DMS) recipients (DMS: 100 microg/ 100 g body wt daily for the first 10 days and ACTH: 0.5 IU / 100 g body wt daily for the next 10 days); (2) Only moderate and higher doses (50 microg/100 microg per 100 g body wt daily for 10 days) of corticosterone increased adrenomedullary activity and blood sugar level and the responses were also dose-dependent. But aldosterone treatment in all doses (same as for corticosterone) had no significant effect on the adrenal medulla or blood sugar level; (3) Only moderate and higher doses of norepinephrine or epinephrine (same as for corticosterone) caused adrenomedullary atrophy with depletions of norepinephrine and epinephrine levels but elevated the glycemic level. The findings are briefly discussed. PMID- 15521653 TI - A comparative study on the biology of Macropis fulvipes (Fabricius, 1804) and Macropis europaea Warncke, 1973 (Hymenoptera: Apoidea: Melittidae). AB - Seasonal and daily activity of Macropis fulvipes (Fabricius, 1804) and M. europaea Warncke, 1973 are given. Their environmental preferences, foraging distances and host plants are presented. Life cycles and nest architecture are described. PMID- 15521654 TI - First European record of Paramecium septaurelia and the discovery of new European habitats of P. pentaurelia and P. sexaurelia in Russia (Astrakhan and Volgograd regions). AB - The presence of several species of the P. aurelia complex was revealed in the studied regions. In the Volgograd region P. primaurelia, P. biaurelia, P. triaurelia, and P. novaurelia were recorded. In the Astrakhan Nature Reserve P. primaurelia, P. pentaurelia, P. sexaurelia, and P. septaurelia were identified. Among these species, P. septaurelia was recorded for the first time in Europe, known before only from the territory of the USA, P. pentaurelia and P. sexaurelia are species rare in Europe. The studied regions are very rich in species of the P. aurelia complex and worthy of future studies. PMID- 15521655 TI - Effect of Pseudomonas aeruginosa crude proteolytic fraction on antibacterial activity of Galleria mellonella haemolymph. AB - The antibacterial activity of immune haemolymph Galleria mellonella directed against Escherichia coli D31 was destroyed by Pseudomonas aeruginosa crude proteolytic fraction. This was demonstrated by diffusion well assay and acid gel electrophoresis and subsequent bioautography. On the contrary, lysozyme activity appeared to be insensitive to extracellular proteases of P. aeruginosa when activity was determined using the bioautography method. In addition, no change in lysozyme protein level was observed by immunoblotting with specific antibodies directed against G. mellonella lysozyme, which confirmed that lysozyme was not degraded by the crude proteolytic fraction of P. aeruginosa. However, a significant decrease of lysozyme activity in naive and immune haemolymph exposed to the action of P. aeruginosa proteins determined by using diffusion well assay was observed. Mechanisms of the observed inhibition require further studies. PMID- 15521656 TI - Long-lasting effects of social stress on peritoneal inflammation in some strains of mice. AB - Adult male mice were kept for one week either one or four animals per cage. Some were maintained under the same social conditions for an additional 9 days (controls); their counterparts were either grouped (4 per cage) or isolated (1 per cage). Changes in housing conditions caused a significant increase of plasma corticosterone measured 30 minutes after separation or grouping of SWISS, C57C3H, and BALB/c but not of C57BL/6 mice. Peritoneal inflammation was induced by i.p. zymosan injection on day 9 after changes in housing conditions when corticosterone was again at its initial level in each group. Peritonitis connected pain symptoms, exudatory PMN numbers, and cytokine (IL-1beta and MPC-1) and corticosterone levels were compared between animals living in stable social conditions with those shifted 9 days earlier from separation to the group or vice versa. These factors were unaffected by social stress in C57BL/6 mice and in SWISS animals transferred from the group to isolation. In all other instances at least two parameters were significantly different in the post-stressed and control animals, being either enhanced or inhibited. In conclusion, social stress had long-term consequences on the course of inflammation in three out of four investigated strains of mice. PMID- 15521657 TI - Effects of rearing density on larval growth and activity of digestive enzymes in Lymantria dispar L. (Lepidoptera: Lymantriidae). AB - Density dependent responses of 4th, 5th and 6th instar gypsy moth larvae were studied at the level of larval mass, midgut loading and activities of three digestive enzymes (alpha-amylase, trypsin and leucine aminopeptidase). High density significantly reduced larval mass while midgut loading (expressed as relative midgut mass) did not change except in the 5th instar where it was increased at high density. Specific amylase and leucine aminopeptidase activities were not affected by crowding. Specific trypsin activity was on average higher in crowded than in isolated larvae. High density also affected the correlations between midgut protein content and activities of two proteolytic enzymes suggesting differences in regulatory mechanisms of insect digestion. The importance of these changes for survival under stressful conditions is discussed. PMID- 15521658 TI - Simultaneous detection of malignant hyperthermia and genetic predisposition for improved litter size in pigs by multiplex PCR-RFLP. AB - The ryanodine receptor gene (RYR1) and the estrogen receptor gene (ESR) are the best commercially used markers for predisposition of stress susceptibility (malignant hyperthermia--MH) and increased litter size, respectively. A simplified method of simultaneous detection of MH and ESR genotypes has been developed. The method is based on simultaneous amplification of fragments of two genes by multiplex PCR and subsequent digestion of the products with two restriction enzymes. The PCR and the digestion could be performed in a single tube and all genotypes could be detected by electrophoretic separation on the same agarose gel. Thus, the development of the method can decrease the cost of the sample analysis and increase the speed and efficiency of the analysis. In our study, frequencies of mutated T allele of the RYR1 gene in Large White (LW), White Meaty (WM) and Landrace (L) were 0.11, 0.13, and 0.15, respectively. Frequencies of the preferred B allele of the ESR gene in the same breeds were 0.35, 0.26, and 0.06, respectively. PMID- 15521659 TI - Random amplified polymorphic DNA fingerprinting as a marker for Paramecium jenningsi strains. AB - The aim of the present study is to establish a common RAPD marker for P. jenningsi using a series of Ro primers and to investigate if strains originating from distant and isolated localities (Japan, China, India, Saudi Arabia) have isolated gene pools and represent distinct species. An analysis of dendrograms constructed on the basis of RAPD-PCR fingerprints with four primers (Ro 460-04, 460-06, 460-07, and 460-10) from the first part of this project (SKOTARCZAK et al. 2004), assigns the strains to two groups consisting of the continental strains (India, Saudi Arabia, China) and Japanese strains that have been considered as a separate sibling species within P. jenningsi. The genetic similarity of the Indian and Arabian strains was ascertained, whereas the Chinese strain formed an independent branch in this sibling species. The primers Ro (460 01,460-02, 460-03, 460-05, 460-08) also distinguish between two groups of strains, although they divide the Japanese strains into two subgroups that are not reproductively isolated. This probably indicates genetic variation within this sibling species. However, it comprises one common gene pool (successful inter-strain crosses) and is reproductively isolated from the other sibling species. The results presented in these papers confirm that the construction of ten band patterns having marker attributes is possible on the basis of DNA amplification from 9 strains of P. jenningsi with the RAPD-PCR fingerprinting method using five primers from the Ro series. The patterns can be assigned to three marker-groups: a general species group, a group differentiating between sibling species, and accessory strain markers. PMID- 15521660 TI - The limitation of DEXA analysis for bone mass determination in mice. AB - An increase in femoral and tibio/fibular bone mass following periosteal membrane stimulation by Moloney sarcoma virus inoculation into thigh muscles of mice was measured in situ on formalin fixed excised hind limbs using a Hologic 4500A Fan Beam X-ray bone densitometer adapted for small bone samples. These results were verified by measurements of constant dry bone mass of the same bones liberated from soft limb tissues by NaOH hydrolysis. There was no consistent data correlation found between the DEXA scan and dry bone mass evaluations. It is concluded that the sensitivity of the DEXA measurement is unsuitable when assessing very small bone samples, weighing merely 20-30 mg. PMID- 15521661 TI - Karyological analysis of two allopatric populations of planarian Polycelis felina (Daly.) in Croatia. AB - The results of preliminary karyological investigations of two geographically distant and morphologically slightly different Polycelis felina (Daly.) populations from central Croatia are reported. The results have shown that individuals of both populations are diploids, with the same chromosome number in their neoblasts (2n=18). Their karyotypes were composed of nine chromosome pairs, three of them are metacentric and the other six are submetacentric. Statistical evaluation of data indicated that both populations of Polycelis felina (Daly.), despite minor differences observed between them, belong to the same karyological biotype. PMID- 15521662 TI - Postoperative care following endoscopic sinus surgery. AB - Postoperative care is very important in the global management of the patient undergoing ESS. Postoperative tamponnade, packing, splinting or stenting is advocated by many surgeons, but its clinical beneficial effects have not clearly been demonstrated. Postoperative rinsing/washing of the surgical cavity offers advantages in healing: high volume, low pressure is to be preferred. Suction cleaning is advocated on a weekly basis starting one week after ESS and continues until secretions, blood, crusts have disappeared. Topical medications have not been the subject of randomized clinical trials. Of the various classes of systemic (oral) medications, only steroids resulted in better short term outcome when using higher doses compared to lower doses. Antibiotics have not shown clinical effects and should not routinely be given. PMID- 15521663 TI - Effects of the nasal strip and dilator on nasal breathing--a study with healthy subjects. AB - OBJECTIVE: To investigate effects of the nose dilating devices on nasal anatomy and breathing in healthy subjects. MATERIALS AND METHODS: 27 healthy subjects were tested when using the Breathe Right nasal strip or the Nozovent dilator. Posterior rhinomanometry, acoustic rhinometry, and a subjective evaluation were used as methods. RESULTS: Both devices significantly increased the minimum cross sectional area of the nasal valve and decreased nasal resistance. The Nozovent dilator proved to be significantly more effective in reducing nasal resistance than the Breathe Right nasal strip. CONCLUSIONS: Nose dilating devices, the Breathe Right nasal strip and the Nozovent dilator, can be used to reduce nasal resistance. More studies are needed to evaluate the usefulness of the devices for patients with chronic obstruction for any reason in the valve area. PMID- 15521664 TI - The role of septal deviation in adult chronic rhinosinusitis: a study of 500 patients. AB - Septal deviation is a common finding and its role in the pathogenesis of chronic rhinosinusitis remains uncertain. The aim of this study was to assess the impact of septal deviation in the region of the ostiomeatal complex with regards to sinus disease, as staged by the Lund-Mackay scoring system in a group of adult patients with symptoms of chronic rhinosinusitis. Five hundred consecutive patients who had undergone CT scanning of the paranasal sinuses for possible chronic sinus disease between Sept. 2002 and Dec. 2003 were recruited. Septal deviation at the ostiomeatal complex on coronal CT scan was evaluated using Radworks diagnostic 5.1 software (Applicare Medical Imaging BU). There were 219 patients with CT positive sinus disease. 281 of the patients had normal sinus CT scans (control group) and were therefore patients with rhinitis rather than rhinosinusitis. Our study showed no significant difference between the chronic rhinosinusitis group and the control group with regards to septal deviation. Nor were we able to demonstrate any correlation between the severity of septal deformity at the ostiomeatal complex region and the severity of sinus disease or OMC disease. PMID- 15521665 TI - The pathogenicity and antibiotic resistance of coagulase-negative Staphylococci isolated from the maxillary and ethmoid sinuses. AB - AIM OF THE STUDY: To investigate the pathogenicity and antibiotic resistance of coagulase-negative staphylococci (CNS) isolated from the maxillary and ethmoid sinuses of patients undergoing endoscopic sinus surgery for chronic sinusitis. PATIENTS AND METHODS: Ninety-three patients (63 males, 30 females) aged between 19 - 68 years, who had undergone functional endoscopic sinus surgery (FESS) for chronic sinusitis, were included in the study. Nasal mucosa, skin and adjacent structures were cleansed with povidone-iodine solution before surgery to prevent a probable contamination. In all patients, nasal swabs were taken before and after the application of povidone-iodine solution. Colonies isolated and identified as Staphylococci in cultures were further investigated for pathogenicity and antibiotic susceptibility. Slime test was used to determine the pathogenicity of CNS. The relationship between antibiotic resistance of pathogenic and non-pathogenic CNS was compared by chi2 analysis. RESULTS: While bacterial growth rate was 62.3% in nasal swab cultures taken before the application of povidone-iodine solution, it decreased to 12.9% after the application of solution. Microorganisms were isolated in 95.6% of cultures taken from maxillary sinuses and in 91.3% of cultures obtained from ethmoid sinuses during the FESS. The most frequently isolated microorganism in each of the sinuses was CNS. Slime test was carried out in 30 CNS isolated. Twelve of these were slime positive and 18 were slime negative. While 83.3% of CNS isolated was resistant to penicilin, all of CNS were sensitive to vancomycin and teikoplanine. The difference between slime positive and slime negative CNS for gentamicin and ciprofloxacin resistance was statistically significant (p<0.05). CONCLUSION: We consider that the pathogenicity tests like slime production and antimicrobial susceptibilities of CNS frequently isolated from the patients with chronic sinusitis should be investigated and also these microorganisms should be kept in mind in the selection of empiric treatment. PMID- 15521666 TI - Point prevalence of allergic rhinitis among Saudi children. AB - BACKGROUND: Study of allergic rhinitis (AR) has only recently started in Saudi Arabia. No estimate of the actual percent of the population suffering from AR, especially children, is available. OBJECTIVE: To study the prevalence of AR in children in the Kingdom of Saudi Arabia, the association with hearing impairment (HI) and bronchial asthma. METHODS: An epidemiological survey was carried out using a modified "International study of asthma and allergies in childhood" (ISAAC) questionnaire for rhinitis phase1. The children with allergic history were subjected to allergy work up. Ten thousand children were surveyed and the clinical history of rhinitis (sneezing, rhinorrhea, irritation, nasal blockage) during the past 12 months was recorded. In total, 9540 children with complete data were interviewed. Demographic characteristics including age, sex, parents' relation, and family history were noted and an Ear, Nose and Throat (ENT) examination and hearing screening was performed. RESULTS: We identified 2529 children with rhinitis (prevalence 26.51%), 25.66% of them with physician diagnosed asthma. Laboratory tests were performed on blood samples from 304 children. A skin prick test with relevant allergens was performed on those aged 6 to 15 years: 61.8% had positive results to one or more allergens, compared to 23.75% in a non-rhinitis control group. HI was found in 450 (17.8%) of the rhinitis group compared to 791 (12.7%) among the non-rhinitis group. CONCLUSIONS: Compared with previous studies in Saudi Arabia, this survey showed higher rates of allergic disease. HI was also found to be higher among rhinitis children. This could be attributed to environmental, social or genetic factors. PMID- 15521667 TI - Invasive fungal rhinosinusitis in immunocompromised patients. AB - INTRODUCTION: Invasive fungal rhinosinusitis (IFR) is one of the most important causes of morbidity and mortality in immunocompromised patients, principally those with cellular immunodeficiency, with mortality ranging from 50 to 80%. Prophylaxis and early diagnosis increase the chances of successful treatment. STUDY DESIGN: Clinical prospective randomized study. AIM: To present cases of IFR and to compare them with data reported in the literature. MATERIAL AND METHODS: Analysis of eleven cases of IFR confirmed by pathologist examination. RESULTS: Aspergillus was found to be the most prevalent pathogen. Symptoms ranged from high fever in most cases to nasal discharge, ulceration of the nasal mucosa, headache and periorbital edema. CONCLUSION: The combination of amphotericin B and endoscopic surgery, associated or not with Caldwell-Luc surgery, showed good results. The use of liposomal amphotericin B also presented a satisfactory outcome. PMID- 15521668 TI - Muscle-building therapy in treatment of nasal valve collapse. AB - OBJECTIVES: The purpose of this paper is to describe the outcome of muscle building therapy for nasal muscles in cases of nasal valve stenosis or collapse. The present study was performed to investigate the best way to combine transcutaneous and intranasal surface electromyography (sEMG) biofeedback training of muscles involved in nasal valve function with a home exercise program and electric stimulation of nasal muscles. METHODS: A randomized pilot study of 3 groups of patients (n1=12, n2=12, n3=10; total 34 patients) presenting with symptoms of obstructed nasal breathing was conducted. All selected patients demonstrated nasal valve stenosis with a positive Cottler maneuver and clinically evident nasal valve collapse. Follow-up ranged from 8 to 12 months. Treatment for Group 1 included transcutaneous and intranasal electric stimulation of nasal muscles only. Treatment for Group 2 included biofeedback training and home exercise program of specific nasal movements, and treatment for Group 3 included surface and intranasal EMG biofeedback assisted specific strategies for nasal muscle education, home exercises and electric stimulation. RESULTS: All patients in these groups exhibited subjective improvement. For Group 3, in 80% the improvement was proved objectively; for Group 2, in 75% the improvement was proved objectively; for Group 1, in 58,33% the improvement was proved objectively. We found no significant difference between the results in Groups 3 and 2 and poorer results in Group 1. CONCLUSION: Relieve of nasal valve stenosis and collapse can be achieved with a complex muscle-building therapy as described. It helps a significant cohort of patients with symptoms of obstructed nasal breathing to avoid surgical intervention. Electric stimulation of the muscles does not contribute significantly in achieving of good results. PMID- 15521669 TI - Histologic evaluation of nasal epithelium of the middle turbinate in untreated OSAS patients and during nCPAP therapy. AB - OSAS-patients complain about nasal disorders. Irritation of the nasal mucosa often leads to termination of nCPAP treatment. The aim of this study is to evaluate whether symptoms are related to histologic changes of the nasal mucosa of the head of the middle turbinate in OSAS-patients. Semi-thin sections of epon embedded middle turbinate biopsy samples from 35 male patients (age 51-75 yr) with OSAS were compared with those of 10 healthy men (age 51-75 yr). In untreated OSAS-patients atrophic epithelium is common whereas ciliated epithelial types are rare. After short-time nCPAP therapy (mean 581h) patches of ciliated epithelium and squamous metaplasia reappear. Short-term nCPAP leads to a partially restoration of the mucosal architecture. During long-time nCPAP therapy (mean 6.737h) squamous metaplasia with conspicuous intraepithelial connective tissue papillae predominates whereas pseudostratified ciliated epithelium is missing. Dense round cell infiltrates in the lamina propria are frequently found. Rhinitic symptoms in OSAS-patients are correlated with marked histological changes of the respiratory epithelium of the head of the middle turbinate. Histological changes in untreated OSAS differ from those of patients during nCPAP-treatment. PMID- 15521670 TI - Rhinology resources on the internet: a critical review. AB - During recent years, the Internet has gradually evolved into a worldwide network, allowing access to vast amounts of information and providing various services. The benefits of medical professionals of using it are growing rapidly, as even more scientific, technical and biomedical resources become available on line. The Internet has always provided useful resources to rhinologists, with the number of relevant sites increasing daily, at a rapid pace. In this article, a review of the information included in several selected rhinology web sites is presented, in order to provide guidance and context for the vast amount of information which is available on the Internet, and to promote the use of the medium to rhinologists who do not have extensive experience in computers or telecommunications. PMID- 15521671 TI - Orbitoethmoid aneurysmal bone cyst. Case report and literature review. AB - Aneurysmal bone cyst is a benign, vascular, variable growing and expansive lesion that can occur in any part of the skeletal system, but mainly in long bones and vertebrae. We present a case of orbitoethmoid aneurysmal bone cyst in a 62-year old female presenting epiphora. Nasal endoscopy was normal. Sinus CTscan revealed an expansive mass in the right ethmoid sinus extending and destroying the right lamina papiracea. The tumor was completely resected through paralateral rhinotomy. Histological analysis showed fibrous septa containing multinucleated giant cells and bone tissue surrounding blood vessel lumens, bordered by endothelial cells. These findings are characteristic of aneurysmal bone cysts. After 30 months of postoperative follow-up the patient remains disease-free and asymptomatic. PMID- 15521672 TI - Nasal teeth associated with rhinosinusitis. AB - Intranasal ectopic dentition is a rare clinical entity. It may be asymptomatic or can be associated with different symptoms. In 25% of the reported cases an association with rhinosinisitis is suspected. The possible pathophysiology, diagnosis and treatment are discussed. Teeth in the floor of both nasal cavities, associated with chronic rhinosinusitis, are reported for the first time, and a new treatment option of endonasal extraction under microscopic control is presented. PMID- 15521673 TI - Juvenile angiofibroma of the maxillary sinus. AB - Primary extra-nasopharyngeal sites of angiofibromas are extremely unusual. We describe a rare case of extra-nasopharyngeal angiofibroma in a one year old child arising from the left maxilla and indirectly involving the lacrimal system. The initial presentation was of a swelling in the region of the left medial canthus. Only four cases of extra-nasopharyngeal angiofibromas in children below the age of two have been described. We review the literature on what is known about extra nasopharyngeal angiofibromas. PMID- 15521674 TI - [The Canadian experience with the SARS outbreak--Israeli lessons to be learned]. AB - The SARS epidemic raised tremendous challenges for the Canadian health system, challenges for which the country was not adequately prepared. Due to the serious consequences of this outbreak in terms of lives lost, medical, social and economical losses, the Canadian government has ordered the formation of a national commission to conduct a comprehensive and impartial investigation of these events. In addition, the Canadian Minister of Health has established the National Advisory Committee on SARS and Public Health with a mandate to "provide an assessment of current public health efforts and lessons learned for ongoing and future infectious disease control". Over a period of several months, these committees have thoroughly investigated all aspects of the events, which have led to this inadequate preparedness. In this article, we summarize the committee's conclusions, which are of direct relevance for the Israeli health care system. Hopefully, these important short and long term lessons, learnt by the Canadian at such a high cost, may be implemented locally, thus enabling better control and prevention of similar future threats caused by emerging and re-emerging infectious diseases. PMID- 15521675 TI - [Pediatric liver transplantation--experience at Schneider Children's Medical Center]. AB - BACKGROUND: Introduction of segmental graft transplantation from living donors and split livers from cadaver donors has led to major advances in liver transplantation (LTx) in children. AIM: To report our initial experience with pediatric LTx performed at our center. METHODS: Data collection on all children undergoing LTx between the years 1996-2003 including the analysis of the graft and patient survivals and reports of complications. RESULTS: Forty LTx were performed in 38 children at the mean age of 6.2 years, including two retransplants. There were 15 whole liver allografts and 25 segmental grafts including: 12 living donor grafts, 5 splits and 8 reduced grafts from cadaver donors. At 40 months mean follow-up period, patient and graft survival were 81% and 72.5%, respectively. There was post-transplant mortality in seven cases--5 children died during the first month and two children passed away after 6 months (recurrent disease) and 14 months (metastatic tumor). Vascular complications included: one early and one late portal vein thrombosis (5%) and six cases of hepatic artery thrombosis (15%). In the latest group, 3 grafts were salvaged by thrombectomy and another 3 children underwent re-transplantation. There were two bile leaks (5%) and 6 bile duct strictures (15%). The bile-duct strictures were successfully corrected by surgery in one child and by transhepatic dilatation in another 4 children. One child remained with intrahepatic strictures in one of the two hepatic segments. CONCLUSIONS: The use of segmental liver allografts enables the performance of pediatric liver transplantation in Israel. Gathered experience and enhanced skills will ensure improved results over time. PMID- 15521676 TI - [Cushing disease: long-term follow-up after transsphenoidal surgery]. AB - BACKGROUND: Cushing's syndrome due to an ACTH-secreting pituitary tumor is associated with serious morbidity and mortality. As there is no definitive medical treatment, surgical removal of the tumor via the transsphenoidal route remains the first choice. Postoperative hypocortisolemia is recognized as the best indicator of cure. OBJECTIVE: To report the postoperative outcome and long term follow-up of patients with surgically treated Cushing disease at the Rabin Medical Center. METHODS: We reviewed the medical records of 27 patients with Cushing disease operated on between the period 1990 and 2003. The same experienced surgeon performed all surgeries. RESULTS: Cushing disease accounted for 15% of all pituitary surgeries in our center. The mean age was 46 years, and the female to male ratio was 25:2. Macroadenomas were found in 19% of cases, and a negative MRI in another 19%. The cure rate was 70% overall and 80% when only microadenomas were considered. There were no major perioperative complications. Four out of 8 surgical failures were re-operated, and three achieved cure. After a mean follow-up period of 5.9 years, there was only one recurrence. CONCLUSION: Our results are in accordance with those reported by others and confirm that in the hands of an experienced neurosurgeon, pituitary surgery constitutes an effective treatment for Cushing disease. PMID- 15521677 TI - [Lumbar epidural anesthesia in a very-low-birth-weight infant]. AB - In this case study two combined, light-general and lumbar epidural anesthetics were administered to a premature neonate for the repair of gastroschisis. The female infant's body weight was 1400 grams (first operation) and 1700 grams (second operation). The epidural catheter was inserted between the low-lumbar vertebrae using the "loss of resistance" for saline technique. Intra- and postoperative analgesia were based on epidural administration of bupivacaine and fentanyl. No systemic opioids were administered. The infant recovered from general anesthesia at the end of both operations. Excellent analgesia was accomplished throughout the peri-operative course except for short periods during and after the second operation that resolved after the epidural catheter was withdrawn by a few centimeters. Neither respiratory or hemodynamic depression, nor bupivacaine toxicity were observed. The anesthetic and analgesic management is presented in the article. Technical aspects of lumbar epidural anesthesia, it's advantages over the caudal approach and dosages of epidural anesthetics in small infants, as well as complications and risks are emphasized. PMID- 15521678 TI - [Comorbid psychiatric symptoms in pathological gamblers: anxiety, depression and substance abuse]. AB - Over the centuries, gambling behaviour has been well known and characterized by the combination of pleasure, luck and competition. Our study explored the relationship between pathological gambling, depression and anxiety. We also explored demographic findings and behavioural patterns of the pathological gamblers. Fourty-seven patients were included in this study and they anonymously completed questionnaires which included demographic findings, the Hamilton depression rating scale and the Hamilton anxiety rating scale. The study results demonstrated a strong correlation between depression, anxiety, substance abuse, and pathological gambling. It also presented lower income and higher anxiety levels associated with a higher tendency for gambling. The subjects suffering from depression and anxiety also showed higher levels of suicidality and other abuse dependencies. In order to confirm these preliminary results larger studies are needed in this field. PMID- 15521679 TI - [Knowledge of stroke among the Israeli population]. AB - BACKGROUND: Stroke and its long-term neurological sequelae can be prevented by management of risk factors and seeking medical care as early as possible following the onset of stroke symptoms. AIMS: This study aimed to investigate the scope of knowledge on stroke, and to assess the perceived risk of stroke among the Israeli population. METHODS AND RESULTS: A telephone survey was conducted of a sample study of 300 men and women, 40 years of age or older, that represent the Hebrew speaking population in Israel of this age range. The best known risk factor for stroke was arterial hypertension, but 24% of the study population could not spontaneously recall any risk factor. Nearly half of the study sample (46%) was aware of the fact that a healthy lifestyle may contribute to stroke prevention, but only 24% mentioned that medical follow-up and control of hypertension, dyslipidemia and diabetes mellitus reduce the risk of stroke. Among interviewed subjects who reported the existence of at least one risk factor of stroke, only 14% recognized that they belong to a group at risk of having a stroke. Awareness of neurological disabilities resulting from stroke was relatively high, with only 15% of responders unable to name any disability. CONCLUSION: Our study demonstrates the lack of knowledge on stroke among the Israeli population. Knowledge is particularly poor regarding the possibility of stroke prevention through risk factor management, and with respect to recognition of symptoms of acute stroke. A program directed at raising the awareness and knowledge of stroke by the Israeli public is required for effective stroke prevention and therapy. PMID- 15521680 TI - [A living will--autonomy in dementive states]. AB - Demented patients may refuse to eat at the end of life. Many caregivers believe that food is as essential as air, and hence, tube feeding is as important as mechanical ventilation. Many believe that demented patients should be fed in any event, even against their will. The aim of this study was to assess the opinions of the elderly about tube feeding and advanced medical procedures in dementia. Inhabitants of protected homes, over 70 years of age, were asked to complete a questionnaire with demographic details, self-estimation, and self grading according to mobility, quality of life, function, pain, family and environmental support. They were asked what they consider to be a situation worse than death, and then, to grade their consent to life support procedures in the different stages of dementia: percutaneous endoscopic gastrostomy, nasogastric tube, resuscitation, artificial respiration or surgery. They were also asked whether they want a living will for medical interventions in dementia for future guardian consideration. One hundred and twenty questionnaires were distributed and 61 were completed (compliance of 50.8%) including responses from 47 women (77%). The average age was 83 years. Most of the participants were women of European origin, not religious, executives with an average of 12 years of education. More than 70% of participants opposed life-supporting procedures in lower stages of dementia, and more than 80% in higher stages. Ninety-five percent of the participants believed in a living will that denied tube feeding and advanced therapies in dementia. PMID- 15521681 TI - [Trichinosis outbreak among Thai immigrant workers in the Hadera sub-district]. AB - INTRODUCTION: Trichinosis also referred to as Trichinelliasis or Trichiniasis occurs sporadically in Israel and is not prevalent. Most reported cases have occurred in foreign born residents who were infected outside of Israel or in foreign workers who have eaten raw or undercooked meat. We report an outbreak of Trichinosis in Thai agricultural workers that occurred in February-March 2002. Of 120 workers in Moshav Sde Yitzchak, 30 became ill with thrichinosis. To the best of our knowledge, this is the largest outbreak of its kind reported in Israel. OBJECTIVES: To alert physicians in hospitals and clinics to consider the possibility of trichinosis in their differential diagnosis in light of the substantial population of foreign born workers in Israel, currently estimated at 238,000. We describe the outbreak and review the symptoms and signs of the disease, as well as the modes of transmission and treatment. RESULTS: Eosinophilia was found in 22 (73.3%) of the cases. The average CPK level in 27 of the cases was 1312. Antibodies to trichinella were positive in the 12 persons who agreed to be tested. CONCLUSION: The demographic changes in Israel over the past years should lead physicians to consider trichinosis in their differential diagnosis when presented with a patient with a suggestive history as well as compatible signs and symptoms. The physician has a legal obligation to report outbreaks of trichinosis to the District Health Office. Health education for at risk populations is indicated. PMID- 15521682 TI - [Pathological gambling in Israel as a medical and social problem]. AB - This article discusses the phenomenon of pathological gambling in Israel today, emphasizing its medical as well as social dimensions. After a brief historical review of pathological gambling in world and Jewish history, certain psychological aspects of the disorder are reviewed. The typology of gamblers is mentioned, and the relationship between gambling and affective disorders is briefly discussed. Some of the forensic implications of pathological gambling are also mentioned. PMID- 15521683 TI - [Endoscopic treatment of vesicoureteral reflux]. AB - OBJECTIVES: Endoscopic subureteral injection of tissue-augmenting substances, a 15-minute outpatient procedure has become an alternative to long-term antibiotic prophylaxis and surgical intervention in the treatment of vesicoureteral reflux (VUR) in children. MATERIALS AND METHODS: We searched MEDLINE using the words: vesicoureteral reflux, treatment, the long-term results of endoscopic treatment of reflux. We summarized the worldwide data regarding endoscopic treatment of VUR using various tissue-augmenting substances presently available. RESULTS: In terms of effectiveness and long-term successful results, polytetrafluoroethylene is still the most reliable injectable material for the endoscopic treatment of VUR. However, Dextranomer/hyaluronic acid copolymer (Deflux) is a new promising tissue augmenting substance which might be able to replace Teflon in the endoscopic treatment of reflux in terms of a similar to Teflon reflux cessation rate and exhibiting no evidence of migration. CONCLUSION: Endoscopic subureteral polytetrafluoroethylene injection is a simple and effective outpatient procedure for in the treatment of vesicoureteral reflux. No long-term morbidity was observed in our patients with small amounts of injectable polytetrafluoroethylene. PMID- 15521684 TI - [Early events in chlamydial infection of host cells]. AB - Chlamydiae, gram-negative, obligate intracellular bacteria, are major pathogens worldwide, causing several diseases including trachoma, respiratory diseases and sexually transmitted disease. Penetration of chlamydiae to epithelial cells, the environment which supports their growth and survival, leads to various events that begin with changes to the bacteria-containing vacuole, allowing for its progression from the endosomal to the exocytic pathway. The changes include fusion with vesicles carrying glycerophospholipids and sphingolipids and originating in the endoplasmic reticulum (ER) and the Golgi apparatus. The bacteria then reproduce in the inclusion vesicles. In this survey we describe the chlamydial life cycle and review recent reports on early intracellular events in chlamydial infection. While antibiotics currently recommended for treatment of chlamydial infections interfere with bacterial macromolecular synthesis, newer forms of treatment may be developed based on our increasing understanding of chlamydial manipulation of intracellular processes. This manipulation, described in this article on early intracellular events in chlamydial infection, enables these pathogens to escape destruction in the endosomal compartment and begin replication in the target cell. PMID- 15521685 TI - [The relationship between attention deficit hyperactivity disorder and sleep alertness problems]. AB - Several studies have reported a relationship between Attention Deficit Hyperactivity Disorder (ADHD) and sleep problems. Parents of children with ADHD report on decreased nocturnal sleep efficiency and sleep fragmentation compared to controls. On the other hand, objective measurements often show that these children sleep longer, and there is even a theory that suggests that daytime somnolence is the primary cause of the inattention problems seen in the disorder. Experimental sleep restriction, which leads to daytime somnolence, has been shown to be associated with ADHD-like behavior and poor cognitive achievements. In a study that we have recently performed, we found that children with ADHD had high prevalence of primary sleep disorders and objective daytime somnolence. This finding can explain why Ritalin, which is a stimulant medication, improves symptoms in these children. Sleep problems, primarily treatable such as Obstructive Sleep Apnea (OSA) or Periodic Limb Movements Disorder in Sleep (PLMD), should be investigated in the assessment of these children. PMID- 15521686 TI - [What causes English sweats?]. AB - English sweating disease also known as Sudor Anglicus is one of the least familiar epidemics of the Middle Ages, striking England 5 times during the 15th and 16th centuries before fading. This article will discuss the knowledge available to us about this fascinating epidemic, its characteristics and causes. PMID- 15521687 TI - [A proper intubation--elementary but not always available]. PMID- 15521688 TI - Perception beyond physics? PMID- 15521689 TI - Visual perception of orientation is categorical near vertical and continuous near horizontal. AB - Four experiments were conducted to examine whether visual-orientation information is perceived categorically. In experiments 1 and 3, adult participants sorted oriented line stimuli into broad oblique and narrow vertical or horizontal categories. Experiments 2 and 4 showed that categorical discrimination of orientation occurred only near the vertical-oblique boundary. The data indicate that there is categorical perception near vertical and more continuous perception near horizontal. The results are relevant to the debate over whether categorical perception is derived from perceptual structure, verbal coding, or within-task learning. In addition, the asymmetrical perception of orientation around vertical and horizontal is consistent with the possibility that there may be differences in the functional significance of orientation near the two main axes. PMID- 15521690 TI - Colour vision brings clarity to shadows. AB - We have revealed a new role for colour vision in visual scene analysis: colour vision facilitates shadow identification. Shadows are important features of the visual scene, providing information about the shape, depth, and movement of objects. To be useful for perception, however, shadows must be distinguished from other types of luminance variation, principally the variation in object reflectance. A potential cue for distinguishing shadows from reflectance variations is colour, since chromatic changes typically occur at object but not shadow boundaries. We tested whether colour cues were exploited by the visual system for shadow identification, by comparing the ability of human test subjects to identify simulated shadows on chromatically variegated versus achromatically variegated backgrounds with identical luminance compositions. Performance was superior with the chromatically variegated backgrounds. Furthermore, introducing random colour contrast across the shadow boundaries degraded their identification. These findings demonstrate that the visual system exploits inbuilt assumptions about the relationships between colour and luminance in the natural visual world. PMID- 15521691 TI - Motion-induced overestimation of the number of items in a display. AB - Subjects were asked to report the number of items in a display as the items moved along a circular path around the fixation point. As the rotation speed increased, the apparent number of items also increased. This motion-induced overestimation (MIO) effect was investigated in three experiments. In the first experiment, the effect of rotation speed and set size was explored with an enumeration task. The overestimation error increased with an increase in speed or number of items in the display. In the second experiment, we used an adjustment paradigm to measure the speed threshold of MIO effect onset. Temporal rate of the display, which was defined as product of rotation speed and the number of rotating items, was the determining factor of MIO onset. In the third experiment, moving items were marked with different colours. Surprisingly, the number of perceived items was still overestimated even though the number of perceived colours was not. PMID- 15521692 TI - Bottom-up and top-down control in visual search. AB - Previous research suggests that the allocation of attention is largely controlled either in a stimulus-driven or in a goal-driven manner. To date, few studies have systematically manipulated variables affecting stimulus-driven and goal-driven selection independently in order to investigate how both manners of control interrelate and affect performance in visual search. In the present study observers were presented with search displays consisting of an array of line segments rotated at various orientations. The task of observers was to indicate the presence or absence of a vertical line segment (the target) presented amongst a series of nontargets and possibly one distractor. By varying the absolute differences in orientation between the target, nontargets, and distractors, relative target-distractor salience and target-distractor similarity were independently manipulated to investigate the contribution of stimulus-driven and goal-driven control. The major result was that relative target-distractor salience and target-distractor similarity affected search performance independently. Performance was better in cases where the irrelevant distractor was not a salient item in the search display and did not look similar to the target. The results are discussed in terms of models of attentional control. PMID- 15521693 TI - Perceptual functions in prosopagnosia. AB - Some patients with prosopagnosia may have an apperceptive basis to their recognition defect. Perceptual abnormalities have been reported in single cases or small series, but the causal link of such deficits to prosopagnosia is unclear. Our goal was to identify candidate perceptual processes that might contribute to prosopagnosia, by subjecting several prosopagnosic patients to a battery of functions that may be necessary for accurate facial perception. We tested seven prosopagnosic patients. Three had unilateral right occipitotemporal lesions, two had bilateral posterior occipitotemporal lesions, and one had right anterior-to-occipital temporal damage along with a small left temporal lesion. These lesions all included the fusiform face area, in contrast to one patient with bilateral anterior temporal lesions. Most patients had impaired performance on face-matching tests and difficulty with subcategory judgments for non-face objects. The most consistent deficits in patients with lesions involving the fusiform face area were impaired perception of spatial relations in dot patterns and reduced contrast sensitivity in the 4 to 8 cycles deg(-1) range. Patients with bilateral lesions were impaired in saturation discrimination. Luminance discrimination was normal in all but two patients, and spatial resolution was uniformly spared. Curvature and line-orientation discrimination were impaired in only one patient, who also had the most difficulty with more basic-level object recognition. We conclude that deficits in luminance, spatial resolution, curvature, line orientation, and contrast at low spatial frequencies are unlikely to contribute to apperceptive prosopagnosia. More relevant may be contrast sensitivity at higher spatial frequencies and the analysis of object spatial structure. Deficits in these functions may impair perception of subtle variations in object shape, and may be one mechanism by which the recognition defect in prosopagnosia can extend to other classes of object subcategorization. PMID- 15521694 TI - Interpersonal perception in Japanese and British observers. AB - We compared performance of Japanese and British observers in deciphering images depicting Japanese interpersonal relationships. 201 Japanese and 215 British subjects were assessed by means of a test consisting of 31 photograph problems accompanied by two or three alternative solutions one of which was correct. Japanese subjects outperformed British subjects on the test overall (z = 3.981, p < 0.001). A two-factor ANOVA (culture x gender) was performed for each of the problems. A cultural effect was found in 17 problems. Surprisingly, British subjects outperformed Japanese subjects in 7 of these problems. There was a gender effect in 4 problems and a culture x gender interaction in 6 problems. The results indicate that cultural experience facilitates nonverbal appraisal of interpersonal relationships, but it may sometimes cause specific errors. Differences in the perceptual cues used suggest that British subjects had difficulty reading Japanese facial expressions. PMID- 15521695 TI - Eye movements during intentional car following. AB - Does intentional car following capture visual attention to the extent that driving may be impaired? We tested fifteen participants on a rudimentary driving simulator. Participants were either instructed to follow a vehicle ahead through a simulated version of London, or were given verbal instructions on where to turn during the route. The presence or absence of pedestrians, and the simulated time of the drive (day or night) were varied across the trials. Eye movements were recorded along with behavioural measures including give-way violations, give-way accidents, and kerb impacts. The results revealed that intentional car following reduced the spread of search and increased fixation durations, with a dramatic increase in the time spent processing the vehicle ahead (controlled for exposure). The effects were most pronounced during nighttime drives. During the car-following trials participants were also less aware of pedestrians, produced more give-way violations, and were involved in more give-way accidents. The results draw attention to the problems encountered during car following, and we relate this to the cognitive demands placed on drivers, especially police drivers who often engage in intentional car following and pursuits. PMID- 15521696 TI - Jitter and size effects on vection are immune to experimental instructions and demands. AB - Both coherent perspective jitter and explicit changing-size cues have been shown to improve the vection induced by radially expanding optic flow. We examined whether these stimulus-based vection advantages could be modified by altering cognitions and/or expectations about both the likelihood of self-motion perception and the purpose of the experiment. In the main experiment, participants were randomly assigned into two groups-one where the cognitive conditions biased participants towards self-motion perception and another where the cognitive conditions biased them towards object-motion perception. Contrary to earlier findings by Lepecq et al (1995 Perception 24 435-449), we found that identical visual displays were less likely to induce vection in 'object-motion bias' conditions than in 'self-motion bias' conditions. However, significant jitter and size advantages for vection were still found in both cognitive conditions (cognitive bias effects were greatest for non-jittering same-size control displays). The current results suggest that if a sufficiently large vection advantage can be produced when participants are expecting to experience self-motion, it is likely to persist in object-motion-bias conditions. PMID- 15521697 TI - Haptic perception of virtual surfaces: scaling subjective qualities and interstimulus differences. AB - We examined, in two experiments, the perceptual scaling of the properties of haptically examined virtual surfaces, and the way in which these properties subjectively combine. Participants used a consistent movement pattern to explore, with a stylus, virtual surfaces generated by a force-feedback device. In experiment 1, four surface properties (bump size, friction, resistance to normal force, and vibration amplitude) were varied individually, in separate blocks of trials. Free magnitude estimates of the subjective dimensions corresponding to these properties showed that all four dimensions conformed closely to the power law, except at very low stimulus values. Exponents for bump size (0.80) and stiffness (1.01) were consistent with values established in earlier work with direct touch of real surfaces. Surprisingly, the exponent for stickiness, not previously measured, was much higher than those for other dimensions (1.49). In experiment 2, dimensional combinations were analyzed by asking subjects to give magnitude estimates of the subjective difference between pairs of surfaces differing in one or two properties. Magnitude estimates of a given one dimensional difference were generally larger when the subject was pressing down firmly on the surfaces, than when only gentle downward pressure was required; this result suggests that forces generated when a surface is haptically examined are interpreted as invariant indicators of the magnitudes of the surface properties themselves. Estimates of one-dimensional differences were also used to make predictions of two-dimensional differences, under assumptions of dimensional integrality and separability. The results fell between these two sets of predictions, indicating only modest integration of surface properties examined with indirect touch. PMID- 15521698 TI - Endothelial dysfunction in patients with peripheral arterial disease and chronic hyperhomocysteinemia: potential role of ADMA. AB - Hyperhomocysteinemia is associated with an enhanced risk for cardiovascular disease. Patients with peripheral arterial disease (PAD) show an increased prevalence of hyperhomocysteinemia. A decreased biological activity of nitric oxide (NO) may contribute to homocysteine-associated endothelial dysfunction. This study was designed to investigate whether elevated levels of the endogenous NO synthase inhibitor asymmetric dimethylarginine (ADMA) are involved in endothelial dysfunction in patients with chronic hyperhomocysteinemia and PAD. A total of 76 patients (58 males and 18 females; mean age 65.2 +/- 2.0 years) with PAD were included in the analysis and characterized according to demographic variables and cardiovascular risk factors. Flow-dependent vasodilation (FDD) was determined by high-resolution ultrasound in the radial artery. Total plasma homocysteine (plasma tHcy) and ADMA levels were measured by HPLC. Urinary nitrate was quantified using gas chromatography-mass spectrometry. Patients with plasma tHcy in the highest tertile (n = 27; i.e. > 10.6 micromol/l) had a mean plasma level of 14.4 +/- 1.21 mol/l compared with 9.9 +/- 0.1 micromol/l in those patients in the middle tertile (n = 22; p < 0.05) and 9.4 +/- 0.1 micromol/l in those in the lowest tertile (n = 27; i.e. <9.6 micromol/l; p < 0.05). The hyperhomocysteinemic individuals (highest tertile) had a significantly decreased FDD compared with healthy age-matched controls (n = 15) (7.6 +/- 1.0 vs 13.0 +/- 0.4%; p < 0.05), higher plasma ADMA concentrations (4.0 +/- 0.3 vs 2.6 +/- 0.3 micromol/l; p < 0.05), and a lower urinary nitrate excretion rate (89.5 +/- 13.4 vs 131.3 +/- 17.9 micromol/mmol creatinine; p < 0.05) compared with patients with plasma tHcy in the lowest tertile. Multivariate regression analysis including plasma tHcy, ADMA, total cholesterol, diabetes mellitus, smoking, and systolic blood pressure revealed ADMA as the only significant factor determining FDD (p < 0.05). In conclusion, we demonstrated a stronger relationship between impaired endothelial function and elevated ADMA levels in comparison with plasma tHcy concentrations in patients with PAD and chronic hyperhomocysteinemia. This may raise the question of whether different therapeutical options that interact indirectly with plasma tHcy, i.e. treatment with ACE inhibitors and AT1-receptor blockers to reduce ADMA plasma concentrations or L-arginine, could be a beneficial tool for treating patients with hyperhomocysteinemia. PMID- 15521699 TI - Endothelial dysfunction and type of cigarette smoked: the impact of 'light' versus regular cigarette smoking. AB - Acute cigarette smoking leads to temporary endothelial dysfunction, which is an early event in atherogenesis. Sufficient data concerning the effect of cigarettes with low tar and nicotine yield are lacking. Seventeen healthy individuals (nine women, eight men, aged 27.8 +/- 3.6 years) were subjected to evaluation of endothelial function by means of endothelium-dependent, flow-mediated dilatation (FMD) of the brachial artery, before, immediately after and 30, 60 and 90 min after smoking a regular cigarette (nicotine 0.9 mg, tar 12 mg) orthe corresponding 'light' cigarette (nicotine 0.6 mg, tar 8 mg). The following day, measurements were repeated after smoking the opposite kind of cigarette. Baseline FMD was 6.1 +/- 1.6% and 7.2 +/- 2.0% in the light and regular cigarette groups, respectively (p = NS). The overall effect of the regular cigarette over time on FMD compared with the light cigarette was significantly different (F = 3.039, p = 0.023). FMD was significantly depressed after smoking both types (light: F = 8.192, p < 0.001; regular: F = 16.698, p < 0.001). Immediately after smoking, FMD declined in both groups (light: 3.0 +/- 2.4% and regular: 1.6 +/- 3.2%, p < 0.001 and p < 0.001, respectively), and it remained significantly depressed in the regular cigarette group at 30 min (0.75+/-1.5%, p < 0.001) and 60 min (3.5 +/- 3.1%, p = 0.024), while in the light cigarette group FMD differences were abolished at 30, 60 and 90 min after smoking. In conclusion, acute smoking of both regular and light cigarettes leads to temporary vasomotor dysfunction; its duration is shorter after smoking a 'light' cigarette. PMID- 15521700 TI - Inflammatory markers, D-dimer, pro-thrombotic factors, and physical activity levels in patients with peripheral arterial disease. AB - Men and women with lower extremity peripheral arterial disease (PAD) have reduced physical activity levels compared with persons without PAD. We describe associations between physical activity levels with D-dimer, pro-coagulant factors, and inflammatory markers in patients with PAD. Participants were 188 patients with PAD identified from non-invasive vascular laboratories. Physical activity was measured over 7 days with a vertical accelerometer. We measured the ankle-brachial index (ABI) and levels of D-dimer, C-reactive protein (CRP), fibrinogen, serum amyloid A (SAA), prothrombin 1.2, t-PA antigen, PAI-1, and the t-PA antigen/PAI-1 ratio. Adjusting for age, sex, race, body mass index, ABI, comorbidities, smoking, total cholesterol/HDL ratio and statin use (for CRP only), we found significant inverse linear associations between physical activity levels and log D-dimer (p = 0.002), log CRP (p < 0.001), fibrinogen (p = 0.014), and log SAA (p = 0.012). There were no significant associations between physical activity levels and other blood factors. In an analysis adjusting for all blood factors simultaneously along with known and potential confounders, log D-dimer was the only blood factor associated significantly with physical activity levels (p = 0.036). Based on these findings, future studies should assess whether interventions to increase physical activity in patients with PAD reduce levels of D-dimer and inflammatory markers. PMID- 15521701 TI - The associations between smoking, physical activity, dietary habits and plasma homocysteine levels in cardiovascular disease-free people: the 'ATTICA' study. AB - Homocysteine levels are considered, by some investigators, as an independent factor of cardiovascular disease; however, others suggest that there is not a causal relationship. The aim of this work was to investigate the associations between homocysteine levels and several lifestyle-related factors. The ATTICA study is a population-based cohort that has randomly enrolled 1128 adult men and 1154 women, stratified by age and gender, from the greater area of Athens, during 2001-2002. Among several demographic, lifestyle, clinical and biochemical characteristics, we measured total plasma homocysteine levels. For the present analysis, we excluded people who had a history of cardiovascular disease. The factors that showed the strongest relationship with homocysteine levels, in both genders, even after adjusting for several potential confounders were: cigarette smoking (p = 0.03), endurance exercise (inverse, p < 0.05), fruit (inverse, p = 0.01) and vegetable intake (inverse, p = 0.04), and alcohol (p = 0.04) and coffee intake (p = 0.04). In conclusion, several lifestyle-related factors seem to be associated with homocysteine levels. However, the vast majority of the associations were weak when we adjusted for other co-factors. The latter may indicate the mutual confounding effect of various socio-demographic, anthropometric and other clinical characteristics on the relationship between homocysteine and cardiovascular risk. PMID- 15521702 TI - Transcutaneous partial pressure of oxygen after surgical wounds. AB - Transcutaneous partial pressure of oxygen (TcPO2) predicts wound healing in critical ischemia. However, no studies have controlled for the effect of wounds on TcPO2. In this study, in 24 hip or knee arthroplasty cases without vascular disease, TcPO2 was measured preoperatively at the incision site, contralaterally, and at a reference site on the chest wall. The measurements were repeated at 2 days and 2 months postoperatively and compared. TcPO2, normal preoperatively, decreased at all sites on the second postoperative day (p < 0.001), with a trend toward a greater decrease at the wound (p = 0.09). All wounds healed, and TcPO2 returned to baseline at all sites 2 months postoperatively. In conclusion, acute surgical wounds are associated with a significant and widespread decrease in cutaneous TcPO2 in the setting of normal arterial perfusion. This decrease is likely multifactorial and deserves further investigation. PMID- 15521703 TI - Common carotid pseudoaneurysm after carotid endarterectomy: a case presentation. AB - A patient with a previous history of neck cancer and carotid endarterectomy presents with a pulsatile cervical mass. Further evaluation reveals the presence of a pseudoaneurysm of the common carotid artery. The endovascular treatment options and techniques employed to exclude the pseudoaneurysm are presented and discussed. PMID- 15521704 TI - Vascular medicine in 2004: major steps toward national prominence. PMID- 15521705 TI - Duplication of the inferior vena cava and abdominal aortic aneurysm. PMID- 15521706 TI - Recurrent swelling of the left supraclavicular fossa. PMID- 15521707 TI - Adherence to a Mediterranean diet and survival in a Greek population. Trichopoulou A, Costacou T, Bamia C, Trichopoulos D. N Engl J Med 2003; 348: 2599 608. PMID- 15521708 TI - Effect of insulin on spontaneous and progesterone-induced GVBD on Bufo arenarum denuded oocytes. AB - Progesterone is considered as the physiological steroid hormone that triggers meiosis reinitiation in amphibian oocytes. Nevertheless, isolated oocytes can be induced to undergo germinal vesicle breakdown (GVBD) in a saline medium by means of treatment with various hormones or inducing agents such as other steroid hormones, insulin or an insulin-like growth factor. It has been demonstrated that Bufo arenarum oocytes obtained during the reproductive period (spring-summer) resume meiosis with no need of an exogenous hormonal stimulus if deprived of their enveloping follicle cells, a phenomenon called spontaneous maturation. This study was undertaken to evaluate the participation of the purine and phosphoinositide pathway in the insulin-induced maturation of oocytes competent and incompetent to mature spontaneously, as well as to determine whether the activation of the maturation promoting factor (MPF) involved the activation of cdc25 phosphatase in Bufo arenarum denuded oocytes. Our results indicate that insulin was able to induce GBVD in oocytes incompetent to mature spontaneously and to enhance spontaneous and progesterone-induced maturation. In addition, high intracellular levels of purines such as cAMP or guanosine can reversibly inhibit the progesterone and insulin-induced maturation process in Bufo arenarum as well as spontaneous maturation. Assays of the inhibition of phosphatidylinositol-4,5 bisphosphate (PIP2) hydrolysis and its turnover by neomycin and lithium chloride respectively exhibited a different response in insulin- or progesterone-treated oocytes, suggesting that phosphoinositide turnover or hydrolysis of PIP2 is involved in progesterone- but not in insulin-induced maturation. In addition, the inhibitory effect of vanadate suggests that an inactive pre-maturation promoting factor (pre-MPF), activated by dephosphorylation of Thr-14 and Tyr-15 on p34cdc2, is present in Bufo arenarum full-grown oocytes; this step would be common to both spontaneous and hormone-induced maturation. The data presented here strongly suggest that insulin initiates at the cell surface a chain of events leading to GVBD. However, our studies point to the existence of certain differences between the steroid and the peptide hormone pathways, although both involve the decrease in intracellular levels of cAMP, the activation of phosphodiesterase (PDE) and the activation of pre-MPF. PMID- 15521709 TI - Meiotic inhibition with different cyclin-dependent kinase inhibitors in bovine oocytes and its effects on maturation and embryo development. AB - Cyclin-dependent kinase inhibitors (CDKIs) such as butyrolactone I (BL-I) and roscovitine (ROS) maintain bovine oocytes blocked at the germinal vesicle (GV) stage. Bohemine (BOH), another CDKI, has been used for oocyte activation. The objective of this study was to determine whether BOH blocks meiosis and to compare its efficiency with other CDKIs (ROS and BL-I). Oocytes were cultured for 24 h in 0, 50, 100 and 150 microM BOH to determine the best concentration for blocking meiosis (experiment 1). GV rates were 3.3%, 64.5%, 83.3% and 88.9% (0,50, 100 and 150 microM, respectively). Experiment 2 compared meiotic inhibition efficiency of BOH (100 microM), ROS (25 microM) and BL-I (100 microM). BL-I presented the highest GV rates (97.5%). BOH and ROS were similar to each other (85.4% and 79.9%, respectively). To assess the reversibility of meiotic inhibition (experiment 3), oocytes underwent in vitro maturation (IVM) for 18 h after the 24 h inhibition. Control oocytes were submitted to IVM for 18 h (C18) or 24 h (C24). Maturation rates were either similar to (ROS and BL-I: 96.0% and 93.6%, respectively) or superior to (BOH, 96.9%) C24 (91.0%). All groups were superior to C18 (82.5%). In experiment 4, oocytes were treated as in experiment 3 and then in vitro fertilized and cultured for 8 days. Blastocyst rates for BL-I (32.3%) were similar to C24 (35.0%), while those for BOH (20.2%) and ROS (24.2%) were inferior. All groups were inferior to C18 (43.4%). The results show that: (a) BOH inhibits meiosis resumption; (b) BL-I is the most effective of the CDKIs tested for blocking meiosis; (c) culture of oocytes with meiosis inhibitors is fully reversible in terms of nuclear maturation but they may either decrease (BOH and ROS) or maintain (BL-I) embryo development rates. PMID- 15521710 TI - Serotonin localization and its functional significance during mouse preimplantation embryo development. AB - Serotonin is a neurotransmitter functioning also as a hormone and growth factor. To further investigate the biological role of serotonin during embryo development, we analysed serotonin localization as well as the expression of specific serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos. The functional significance of serotonin during the preimplantation period was examined by studying the effects of serotonin on mouse embryo development. Embryo exposure to serotonin (1 microM) highly significantly reduced the mean cell number, whereas lower concentrations of serotonin (0.1 microM and 0.01 microM) had no significant effects on embryo cell numbers. In all serotonin treated groups a significant increase in the number of embryos with apoptotic and secondary necrotic nuclei was observed. Expression of serotonin 5-HT1D receptor mRNA in mouse oocytes and preimplantation embryos was confirmed by in situ hybridization showing a clearly distinct punctate signal. Immunocytochemistry results revealed the localization of serotonin in oocytes and embryos to the blastocyst stage as diffuse punctate cytoplasmic labelling. It appears that endogenous and/or exogenous serotonin in preimplantation embryos could be involved in complex autocrine/paracrine regulations of embryo development and embryo-maternal interactions. PMID- 15521711 TI - Effects of transferring in vitro-cultured rabbit embryos to recipient oviducts on mucin coat deposition, implantation and development. AB - A mucin coat is deposited on rabbit embryos during passage through the oviduct; rabbit blastocysts cultured from the 1-cell stage in vitro have no mucin coat. When cultured blastocysts are transferred to recipients, the lack of mucin coat might account in part for subsequent failure of pregnancy. We have investigated the possibility that mucin coat deposition is induced following transfer of in vitro 72 h-cultured blastocysts to oviducts of asynchronous or synchronous recipients. One-cell embryos were collected by flushing oviducts 19-20 h post coitus and were cultured in vitro for 72 h until they reached the blastocyst stage. The blastocysts were transferred to the oviducts of recipients that were synchronized either with the donors (synchronous) or 1 day later than the donors (asynchronous). They were recovered after 24-48 h and the mucin coat thickness and embryo degeneration rate were measured. The degeneration rate of blastocysts recovered from uteri of synchronous recipients was higher than that from asynchronous recipients (72.2% vs 40.0%). The mucin coats around embryos recovered from oviducts of asynchronous recipients after 48 h were thicker than those from synchronous recipients. More asynchronous recipients were pregnant and gave birth to more pups than synchronous recipients. These results indicate that the oviducts of asynchronous recipients secreted more mucin around the transferred embryos, causing higher rates of implantation of the in vitro cultured blastocysts. PMID- 15521712 TI - Characterization of mid-spindle microtubules during furrow positioning in early cleavage period zebrafish embryos. AB - We report evidence to suggest that during the first few meroblastic cell divisions in zebrafish embryos a dynamic population of central-spindle microtubules serve a crucial function in positioning the cleavage furrow at the surface of the blastoderm. Originating from the mid-zone of the mitotic spindle they develop into what we term a mid-spindle 'pre-furrowing microtubule array' that expands upward and outward from the spindle mid-zone towards the blastodisc surface. We suggest that this structure transmits positional information to the blastodisc cortex that results in the correctly positioned assembly of the cytokinetic contractile apparatus. We also propose that the pre-furrowing microtubule array then develops into a furrow-ingression microtubule array that helps direct and assemble the deepening furrow as it cuts its way through the blastodisc. Due to the location of its origin, the pre-furrowing microtubule array serves to successfully separate the daughter nuclei and thus equally divide the blastoderm. Furthermore, co-localization with elements of the cortical endoplasmic reticulum and their inositol 1,4,5-trisphosphate receptors suggests that the pre-furrowing microtubule array may also play a role in organizing localized Ca2+ transients that have been shown to be essential to the furrow positioning, propagation and deepening process during cytokinesis in zebrafish embryos. PMID- 15521713 TI - Growth factors protect in vitro cultured embryos from the consequences of oxidative stress. AB - The aim of the study was to evaluate the effect of insulin-like growth factors (IGF1 and IGF2), stem cell factor (SCF) and epidermal growth factor (EGF) on the development of embryos exposed to oxidative stress. C3B6F1 female mice were stimulated with 5 IU of pregnant mare serum gonadotropin and 5 IU of equine chorionic gonadotropin (eCG). Two-cell embryos were flushed out from the fallopian tubes 40 h after eCG administration and mating with DBA males. In each experiment embryos were divided into three groups and cultured in (1) control medium, (2) control medium with 0.1 mM hydrogen peroxide and (3) control medium with hydrogen peroxide and separately with IGF1, IGF2, SCF or EGF in concentrations of 1 ng/ml, 10 ng/ml and 100 ng/ml. Under phase-contrast microscopy, 8-cell and compacted embryos, and early, expanded, hatched and outgrown blastocysts were counted at 24 h. The total blastocyst (TB) and inner cell mass (ICM) cell numbers were established by differential staining. Blastocyst cell viability was examined under fluorescence microscopy. To detect apoptosis, TUNEL was performed and visualized under a laser scanning confocal microscope. Hydrogen peroxide decreased embryo growth, blastocyst rates, blastocyst cell viability as well as TB and ICM counts. The TUNEL reaction revealed significantly more apoptotic cells in oxidative stress conditions. Tested factors revealed a varying extent of protective activity against oxidative stress caused by hydrogen peroxide. In media containing hydrogen peroxide and one of the four tested factors (IGF1, IGF2, SCF or EGF) the embryos developed faster than in media with hydrogen peroxide alone. IGF1, IGF2 and EGF increased both TB and (or) ICM counts in embryos exposed to hydrogen peroxide. All tested factors reduced the number of apoptotic cells (TUNEL) in embryos exposed to hydrogen peroxide. PMID- 15521714 TI - Spindle dynamics in living mouse oocytes during meiotic maturation, ageing, cooling and overheating: a study by polarized light microscopy. AB - A liquid crystal polarized light microscope (LC PolScope) was used to examine spindle dynamics in living mouse oocytes. Immature oocytes were cultured for 0-48 h and spindles were imaged with the PolScope at various time points of culture. Oocytes at metaphase I (M-I) and metaphase II (M-II) were also exposed to shifts of temperature from 25 to 41 degrees C to examine the effects of fluctuations of temperature on spindle dynamics. After examination with the PolScope, some oocytes were fixed and examined by immunocytochemical staining and confocal microscopy. After culturing for 6 h, 76% and 2% of the oocytes reached M-I and M II stages and all oocytes had birefringent spindles. When the oocytes were cultured for 14-16 h, 88% and 6% of oocytes were at M-II and M-I stages respectively and all oocytes had birefringent spindles. However, when the oocytes were cultured for 22-48 h, the proportions of oocytes with birefringent spindles decreased as culture time was increased. Exposure of oocytes to 25 degrees C induced spindle disassembly within 10-20 min in both M-I and M-II oocytes. Most (93-100%) oocytes reassembled spindles after warming at 37 degrees C. Furthermore, exposure of oocytes at M-I stage but not at M-II stage, to 30 degrees C also induced significant microtubule disassembly. However, exposure of oocytes to 38-41 degrees C did not obviously change the quantity of microtubules in the spindles, which was measured by retardance. This study indicates that the PolScope can be used to examine spindle dynamics in living oocytes, and it has the advantage over the routine fluorescence microscope in that images can be obtained in the same individual oocyte and the quantity of microtubules can be measured by retardance in living oocytes. These results also indicate that the M II spindle in mouse oocytes is sensitive to oocyte ageing and cooling, but not heating, and M-I spindle is more sensitive to temperature decline than M-II spindle. PMID- 15521715 TI - Glycoconjugate profiles of the lancelet (Branchiostoma lanceolatum) ovary: a lectin histochemical study by laser confocal microscopy. AB - The presence and the distribution of carbohydrate moieties in ripe lancelet (Branchiostoma lanceolatum) oocytes (mean diameter 130 microm) was studied by lectin histochemistry in combination with enzyme and chemical treatments. Binding sites for eight lectins with specificities towards different glycan moieties were studied on sections of the whole body of mature female lancelets. Only three of the lectins tested reacted positively. Concanavalin-A (ConA)-binding glycoconjugates were localized in the cytoplasm, namely in yolk granules, whereas Artocarpus integrifolia (AIA) and Ricinus communis (RCA) agglutinins bound strongly to extracellular coats of the oocyte identified as the jelly coat and vitelline layer. No other tissues of the lancelet body were found to be positive to any lectin tested, except gut enterocytes which reacted strongly with AIA. Reactivity to ConA was abolished by pretreatment of sections with N-glycosidase F but not by mild alkaline hydrolysis, confirming that the glycoconjugates were of the N-linked type. On the contrary, chemical removal of O-linked chains by mild alkaline hydrolysis abolished AIA and RCA reactivity but had no effect on ConA positivity. PMID- 15521716 TI - Effect of GnRH injection timing in the production of pronuclear-stage zygotes used for DNA microinjection. AB - This study was aimed at developing a hormonal treatment protocol in order to optimize the proportion of pronuclear-stage embryos to be used for DNA microinjection in a goat transgenic founder production programme. A total of 46 adult BELE and 47 adult standard goats (1-5 years old) were used as donors and recipients, respectively. They were heat-synchronized using intravaginal sponges containing 60 mg medroxyprogesterone acetate for 10 days with an injection of 125 microg cloprostenol on the morning of the eighth day. Recipients were injected with 400 IU eCG at the time of sponge removal while donors received a total of 133 mg NIH-FSH-P1 (Folltropin-V) given twice daily in decreasing doses over 3 days starting 48 h before sponge removal. Ovulation was induced in donors by injecting 100 microg of GnRH at 24 h (GnRH24) or 36 h (GnRH36) after sponge removal. Embryo recovery was performed by oviduct flushing following a standard mid-ventral laparotomy procedure. The proportion of embryos in the pronuclear stage of development was higher in the GnRH36 group (90% vs 34%, p < 0.01). Embryos were microinjected with a DNA expression cassette followed by transfer to the oviduct of synchronized recipients. A higher, yet not statistically significant, pregnancy rate was found in the recipients transferred with pronuclear-stage embryos compared with those transferred with 2-cell-stage embryos (64% vs 37%, chi-square p = 0.06). One transgenic female founder was produced from the group of recipients transferred with pronuclear-stage microinjected embryos. PMID- 15521717 TI - Effects of sperm concentrations and culture media on fertilization and development of in vitro matured pig oocytes. AB - This study was carried out to investigate the effects of sperm concentrations and culture media on fertilization and development of in vitro matured pig oocytes. The concentrations of frozen-thawed sperm were 0.2 x 10(7), 2 x 10(7), 20 x 10(7) and 200 x 10(7)/ml, respectively. Culture media were NCSU-23, HEPES-buffered (25 mM) NCSU-23, PZM-3 and PZM-4, respectively. Increasing the sperm concentration from 0.2 x 10(7) to 2 x 10(7)/ml, significantly increased the penetration rate. Also, increasing the sperm concentration from 20 x 10(7) to 200 x 10(7)/ml increased the penetration rate from 62.1% to 69.9%, respectively, with no differences between these two concentrations. A similar pattern was observed for polyspermic penetration and male pronucleus formation. The mean number of sperm per oocyte significantly increased in the 20 x 10(7)/ml and again in the 200 x 10(7)/ml sperm concentrations. The percentage of blastocysts from cleaved oocytes at the 2 x 10(7)/ml sperm concentration was significantly higher than that at the 0.2 x 10(7), 20 x 10(7) and 200 x 10(7)/ml sperm concentrations. The percentage of blastocysts from cleaved oocytes and the cell numbers per blastocyst were significantly higher in the HEPES-buffered NCSU-23 culture medium than in the NCSU-23, PZM-3 and PZM-4 culture media under a gas atmosphere of 5% CO2 in air. PMID- 15521718 TI - Characteristics of invasive cells found in between zona pellucida and oocyte during follicular atresia in mice. AB - During the process of follicular atresia, cells are observed to invade the zona pellucida (invasive cells) where they presumably play an important role in eliminating degraded oocytes. Although our preliminary studies have suggested that these cells may originate from granulosa cells and not from macrophages, a detailed morphological analysis of the cells has not been conducted. The objective of this study was to characterize the cells more precisely by electron microscopy and immunohistochemistry, using sexually immature mice. The results show that the invasive cells were first observed within advanced primary (non antral) atretic follicles. The cells frequently contained cytoplasmic lysosome like granules after passing through the zona pellucida. F4/80 and Mac-1, reported as macrophage-specific antibodies, were reactive with the cells in most cases, but some immunonegative invasive cells were also observed. The ultrastructural features of the invasive cells were quite similar to those of granulosa cells, not macrophages. Gap junctions, which are typical cytoplasmic structures of epithelial cells, were frequently identified between neighbouring cells. Although direct evidence indicating a contribution by the cells to the elimination of degenerated oocytes was not obtained, our results strongly suggest that the invasive cells originated from granulosa cells surrounding the zona pellucida, and that they may have a macrophage-like cell function for the elimination of oocytes from atretic follicles in mice. PMID- 15521719 TI - Th1/Th2 CD4+ T cell responses against NY-ESO-1 in HLA-DPB1*0401/0402 patients with epithelial ovarian cancer. AB - The NY-ESO-1 antigen is expressed in a significant proportion of patients with epithelial ovarian cancer (EOC) and appears to be an ideal target for immunotherapy. In order to elucidate the nature of the HLA-DPB1*0401/0402 (DP4+) restricted CD4+ immune response in patients with NY-ESO-1-expressing EOC, peripheral blood CD4+ T cells from HLA-DP4+ patients were stimulated with the NY ESO-1 epitope 157-170 and tested for the release of type 1 (IFN-gamma) and type 2 (IL-5) cytokines in enzyme-linked immunospot assays. Of 14 DP4+ EOC patients who tested seronegative for NY-ESO-1, 3 patients had a detectable CD4+ T cell response to NY-ESO-1 epitope 157-170 by IFN-gamma ELISPOT assay. Six of 10 DP4+ EOC patients with serum antibodies to NY-ESO-1 had CD4+ T cell responses to NY ESO-1 epitope 157-170 by IFN-gamma assay. Six patients had mixed Th1/Th2 CD4+ T cell responses to NY-ESO-1 epitope 157-170 regardless of their antibody response to NY-ESO-1. Four EOC patients had Th1 cells expressing IFN-gamma, but not IL-5. This suggests that the NY-ESO-1 epitope 157-170 stimulates both Th1 and Th2 type CD4+ T cell responses in EOC patients. These data suggest the NY-ESO-1 epitope 157-170 has a key role in the induction of cellular and humoral immune responses against NY-ESO-1-expressing EOC tumors. Our study supports the relevance of cancer vaccine trials with the NY-ESO-1 epitope 157-170 in HLA-DP4+ EOC patients with NY-ESO-1-expressing tumors and strategies to improve Th1-dominated tumor reactive CD4+ T cell bias. PMID- 15521720 TI - Dioxygenation of human serum albumin having a prosthetic heme group in a tailor made heme pocket. AB - Human serum albumin (HSA) is the most abundant plasma protein in our bloodstream and serves as a transporter for small hydrophobic molecules such as fatty acids, bilirubin, and steroids. Hemin dissociated from methemoglobin is also bound within a narrow D-shaped cavity in subdomain IB of HSA. In terms of the general hydrophobicity of the alpha-helical pocket, HSA potentially has features similar to the heme-binding site of myoglobin (Mb) or hemoglobin (Hb). However, the reduced ferrous HSA-heme complex is immediately oxidized by O2, because HSA lacks the proximal histidine that enables the heme group to bind O2. In this paper, we report the introduction of a proximal histidine into the subdomain IB of HSA by site-directed mutagenesis to construct a tailor-made heme pocket (I142H/Y161L), which allows a reversible O2 binding to the prosthetic heme group. Laser flash photolysis experiments revealed that this artificial hemoprotein appears to have two different geometries of the axial-imidazole coordination, and these two species (I and II) showed rather low O2 binding affinities (P1/2O2 = 18 and 134 Torr) relative to those of Mb and Hb. PMID- 15521721 TI - A genetically encoded photocaged amino acid. AB - We have developed a second orthogonal tRNA/synthetase pair for use in yeast based on the Escherichia coli tRNALeu/leucyl tRNA-synthetase pair. Using a novel genetic selection, we have identified a series of synthetase mutants that selectively charge the amber suppresor tRNA with the C8 amino acid, alpha aminocaprylic acid, and the photocaged amino acid, o-nitrobenzyl cysteine, allowing them to be inserted into proteins in yeast in response to the amber nonsense codon, TAG. PMID- 15521722 TI - Rationally designed ligands for asymmetric iridium-catalyzed hydrogenation of olefins. AB - A new class of chiral N,P-ligands for the Ir-catalyzed asymmetric hydrogenation of aryl alkenes has been developed. These new ligands proved to be highly efficient and tolerates a broad range of substrates. The enantiomeric excesses are in the range of the best ever reported. The results can be rationalized with the proposed selectivity model. PMID- 15521723 TI - Fluorescence lifetimes and emission patterns probe the 3D orientation of the emitting chromophore in a multichromophoric system. AB - In this Communication, we report on the fluorescence behavior of an individual first generation multichromophoric dendrimer. The fact that each of the chromophores in time acts as a fluorescent trap is demonstrated by directly probing the dipole orientation of the emitting chromophore by means of defocused wide-field imaging and comparing experimental and calculated emission patterns. It is shown that in such cases the electromagnetic boundary condition effect results in discrete changes in the fluorescence lifetime as a function of time for individual dendrimers. PMID- 15521724 TI - Banana-shaped liquid crystals with two oligosiloxane end-groups: field-induced switching of supramolecular chirality. AB - The first bent core mesogen carrying branched oligosiloxane units at both ends was synthesized and investigated with polarized light microscopy, DSC, X-ray diffraction of well aligned samples, and electrooptical methods. Two different antiferroelectric switching liquid crystalline phases were found. Both can be regarded as modulated smectic phases in which the molecules have extremely large tilt angles. In both mesophases, the field-induced switching takes place by rotation around the molecular long axes. This field-induced reorientation switches the layer chirality. The transition between the two mesophases is associated with a change of the tilt direction from synclinic to anticlinic. The synclinic high-temperature phase is an obliquelike ribbon phase whereas the anticlinic low-temperature phase represents a sinusoidal undulated layer structure (SmCAPA). It is proposed that the phase transition from the ribbon phase to the undulated layer structure is mainly driven by the change of the interlayer correlation. PMID- 15521725 TI - 3,3,3-trichloropropyl-1-triphenylphosphorane: a reagent for the synthesis of (Z) 1,3-enynes, (Z,Z)-1-chloro-1,3-dienes, and 1,3-diynes. AB - 3,3,3-Trichloropropyl-1-triphenylphosphonium chloride is conveniently prepared from 2-chloroethanol, triphenylphosphine, and trichloroacetic acid. Deprotonation of this reagent generates 3,3,3-trichloropropyl-1-triphenylphosphorane, which reacts with aldehydes to give trichloromethylated (Z)-olefins, which are useful for the synthesis of (Z)-1,3-enynes, (Z,Z)-1-chloro-1,3-dienes, and 1,3-diynes in high yields and stereospecificities. PMID- 15521726 TI - Halide-induced supramolecular ligand rearrangement. AB - A novel reaction involving the halide-induced rearrangement of ligands within supramolecular Rh(I) complexes containing hemilabile ligands is presented. Three analogous bis- and trishemilabile ligands have been synthesized to construct bi- and trimetallic Rh(I) macrocyclic complexes. An intentionally added halide source results in the formal rotation of only one hemilabile ligand along the axis that is perpendicular to the plane defined by the aryl backbone of the hemilabile ligands. X-ray structures, as determined by X-ray crystallography, of key intermediates and products are presented. PMID- 15521727 TI - Photochemical attachment of organic monolayers onto H-terminated Si(111): radical chain propagation observed via STM studies. AB - Photochemical reactions of terminal alkenes with hydrogen-terminated silicon surfaces are being used by many groups to produce covalently attached organic monolayers with a wide range of terminal functionalities. Despite the considerable activity in this area, the mechanism for these reactions has not been definitively established. Here we present STM and HREELS data on a sequence of partially reacted samples, showing the progress of the reaction. The attachment reaction is found to proceed via formation of irregularly shaped islands that appear to grow by a pseudorandom walk process. These data support a radical chain propagation mechanism previously suggested for this reaction. However, since the photons employed here (447 nm) lack sufficient energy for Si-H bond cleavage, an alternate mechanism for initiating the chain reaction appears to be required. PMID- 15521728 TI - Absorption and fluorescence spectra of uracil in the gas phase and in aqueous solution: a TD-DFT quantum mechanical study. AB - Here we present the first computations of fluorescence spectra in aqueous solution at an accurate quantum mechanical level. From a methodological point of view, our study shows that by only taking into account both bulk effects and explicit solvent molecules it is possible to reproduce solvent effects on the energy and the intensities of the electronic spectra, especially for what concerns pi/pi* transition. The computed absorption and fluorescence spectra are in a good agreement with the available experimental results. The energy ordering between the lowest energy n-pi* and the pi/pi* transitions in uracil strongly depends on the nature of the embedding medium. The geometry of the first solvation shell is remarkably sensitive to the specific electronic state, suggesting that solvent degrees of freedom can act as S1/S2 coupling modes. PMID- 15521729 TI - Studying nanoparticle-induced structural changes within fatty acid multilayer films using sum frequency generation vibrational spectroscopy. AB - The nonlinear optical technique of sum frequency generation (SFG) vibrational spectroscopy has been used for the first time to study CdS nanoparticle/arachidic acid multilayer structures. Using a combination of per-deuterated and per protonated arachidic acid, it is possible to study individual layers anywhere within the film, buried or on the surface. Before reaction with H2S all layers are highly ordered, but after the reaction the layers become highly disordered, except for the surface layer, which remains well ordered. This sheds new light on the structure and stability of these films and shows that SFG can provide unique structural information. PMID- 15521730 TI - Zeolite nanoclusters coated onto the mesopore walls of SBA-15. AB - Ultrahigh-field 27Al MAS and MQMAS NMR and 129Xe NMR were used to characterize the zeolite-coated SBA-15 materials. This study shows evidence that zeolite nanoclusters are coated on the mesopore surface of SBA-15. These techniques are useful for the detection of zeolite nanoclusters and different aluminum environments in zeolite/aluminosilicate composites, which are difficult or even impossible to detect by conventional techniques. PMID- 15521731 TI - On the origin of dinitrogen hydrogenation promoted by [(eta5 C5Me4H)2Zr]2(mu2,eta2,eta2-N2). AB - The origin of the hydrogenation of the dinitrogen ligand in [(eta5 C5Me4H)2Zr]2(mu2,eta2,eta2-N2) has been investigated by a combined computational and experimental study. Density functional theory calculations on the zirconocene dinitrogen complex demonstrate significant imido character in the zirconium nitrogen bonds, arising from effective pi-back-bonding from the low-valent zirconium and the side-on bound N2 ligand. The twisted ground-state structure of the N2 complex is a key requirement for nitrogen hydrogenation, as calculations on the model complex [(eta5-C5H5)2Zr]2(mu2,eta2,eta2-N2) reveal reduced overlap as the dihedral angle between the zirconocene wedges approaches 0 degrees . Experimentally, isotopic labeling studies on the microscopic reverse are consistent with a 1,2-addition mechanism for nitrogen hydrogenation. PMID- 15521732 TI - Synthesis and single-molecule studies of a well-defined biomimetic modular multidomain polymer using a peptidomimetic beta-sheet module. AB - In the pursuit of advanced biomaterials with combined strength, toughness, and elasticity, a new class of well-defined modular polymers has been synthesized, and their nanomechanical properties have been studied using atomic force microscopy. These polymers are based on a peptidomimetic beta-sheet-based double closed loop (DCL) module, which was designed to overcome the limitation of the modular polymers we reported previously (J. Am. Chem. Soc. 2004, 126, 2059). Single-molecule force-extension experiments revealed the sequential unfolding of these modules as the polymer is stretched, resulting in more regular sawtooth patterned curves similar to those seen in titin and other biopolymers. The single molecule data agreed well with computer modeling, which suggested that hydrogen bonding and pi-stacking are both involved in the formation of small DCL clusters along the polymer chain. PMID- 15521733 TI - The effect of molecular crowding with nucleotide length and cosolute structure on DNA duplex stability. AB - The thermodynamics of DNA duplex structures in the presence of high concentrations of cosolutes in solution were investigated to discern nucleic acid structures and functions in living cells. In the presence of ethylene glycol (EG) and poly(ethylene glycol) (PEG) (MW = 200-8000), the stability of the oligomer DNA duplexes with differing nucleotide length varied, depending on the nucleotide length as well as the size of PEG. It was also revealed that the decrease of water activity is the primary factor for destabilization of the short (8-mer) duplex by addition of high molecular weight PEGs as well as low molecular weight PEGs and other low molecular weight cosolutes. In addition, the number of water molecules taken up per base pair formation was the same for all the PEGs and for 1,2-dimethoxyethane, which was greater than in the cases of glycerol, EG, 1,3 propanediol, and 2-methoxyethanol, suggesting that the solvation of nucleotides may differ, depending on the cosolute structure. These findings are useful not only for understanding nucleic acid structures and functions in cells but also for the design of oligonucleotides applicable for cells, such as antisense nucleic acids, RNAi, and DNA chips. PMID- 15521734 TI - DFT prediction and experimental observation of substrate-induced catalyst decomposition in ruthenium-catalyzed olefin metathesis. AB - Ruthenacyclobutane decomposition, involving competitive beta-hydride transfer to Ru and reductive olefin elimination during ruthenium-catalyzed olefin metathesis, is predicted by density functional theory calculations and experimentally confirmed by propene and butene formation during degenerate Ru-methylidene catalyzed metathesis of ethylene. The results provide new focus on the nature of ruthenium metathesis catalyst decomposition under catalytic conditions. PMID- 15521735 TI - Large structural modification with conserved conformation: analysis of delta(3) fused aryl prolines in model beta-turns. AB - For the first time, the influence of a fused Delta3-arylproline on peptide conformation has been studied by the synthesis and comparison of the conformations of peptides containing proline and pyrrolo-proline, 3 (PyPro). Pyrrolo-proline was demonstrated to be a conservative replacement for Pro in model beta-turns, 4 and 5, as shown by their similar DMSO titration curves, cis/trans-isomer populations, and NOESY spectral data. Pyrrolo-proline may thus be used for studying the structure activity relationships of Pro-containing peptides with minimal modification of secondary structures. PMID- 15521736 TI - One-pot synthesis of high-quality zinc-blende CdS nanocrystals. AB - This paper reports a one-pot synthetic method for producing CdS nanocrystals. We have demonstrated that the nanocrystal nucleation and growth stages can be automatically separated in a homogeneous system with the presence of nucleation initiators. Accelerators used for more than 70 years in rubber vulcanization (i.e., tetraethylthiuram disulfides, and 2,2'-dithiobisbenzothiazole) were found to be effective nucleation initiators for CdS nanocrystal synthesis. The as prepared CdS nanocrystals are highly monodisperse and possess a zinc blende crystal structure. The quantum yield of the band-gap photoluminescence is up to 12% when the surface-trap emission was totally eliminated after a gentle oxidation under laboratory fluorescent light. PMID- 15521737 TI - Three-component coupling based on the "cation pool" method. AB - Sequential one-pot three-component coupling reactions have been developed based on the "cation pool" method. An N-acyliminium ion generated by the "cation pool" method adds to an electron-rich carbon-carbon double bond, such as enamine derivatives and vinyl sulfides, to form the second "cation pool". The addition of nucleophiles such as allylsilanes, enol silyl ethers, Grignard reagents, and organoaluminum compounds led to the formation of the corresponding three component coupling products. PMID- 15521738 TI - Versatile organic (fullerene)-inorganic (CdTe nanoparticle) nanoensembles. AB - Novel organic (positively charged fullerene)-inorganic (negatively charged CdTe nanoparticle) nanoensembles were devised through electrostatic interactions and probed as versatile donor-acceptor hybrids. Photoirradiation of their homogeneous solutions, containing the electrostatically packed components, let to very long lived (1.3 ms) charge separated states. PMID- 15521739 TI - Preparation of benzolactams by Pd(OAc)2-catalyzed direct aromatic carbonylation. AB - We developed a new method for Pd(II)-catalyzed direct aromatic carbonylation in a phosphine-free catalytic system using Pd(OAc)2 and Cu(OAc)2 in an atmosphere of CO gas containing air. The carbonylation proceeded with ortho-palladation, inducing a remarkable site selectivity to afford a variety of five- or six membered benzolactams from secondary omega-arylalkylamines, such as N alkylbenzylamines or N-alkylphenethylamines. PMID- 15521740 TI - Sequential coupling of zincated hydrazone, alkenylboronate, and electrophile that creates several contiguous stereogenic centers. AB - A zincated N,N-dimethylhydrazone of a ketone undergoes stereospecific syn addition to E- or Z-alkenylboronate to generate a gamma-Zn/B dimetallic intermediate, which reacts with a carbon electrophile to give a gamma borylhydrazone in good yield with excellent diastereoselectivity, creating two to four contiguous stereogenic centers in a one-pot reaction. PMID- 15521741 TI - Enhanced catalytic activity of hemoglobin in organic solvents by layered titanate immobilization. AB - A simple layered titanate immobilization can significantly enhance the Hb catalytic activity in organic media relative to native Hb, especially in hydrophilic dioxane systems, where the composites have an about 100 times enhancement. The interlayer water should be responsible for the increased nonaqueous catalytic activities of the immobilized Hb due to its strengthened resistance to the distorting of the Hb surface essential water layer by organic solvents. PMID- 15521742 TI - Simple synthesis route to monodispersed SBA-15 silica rods. AB - A simple, flexible, reproducible, high-yielding (ca. 100%) synthesis of hexagonally facetted SBA-15 straight rods with uniform sizes (ca. 1.5 x 0.4 mum) under static conditions without using inorganic salts is provided. This method affords rodlike particles with temperature-dependent pore sizes ranging from 5.8 to 12.5 nm but with similar external dimensions. This work shows that in order to prepare short monodispersed SBA-15 rods, (i) the absence of stirring is essential, (ii) the temperature of the first stage should not exceed 60 degrees C, and (iii) addition of inorganic salts is not required. PMID- 15521743 TI - Dissolution and regeneration of Bombyx mori silk fibroin using ionic liquids. AB - In this work, the suitability of imidazolium-based ionic liquid solvents is investigated for the dissolution and regeneration of silkworm (Bombyx mori) silk. Within an ionic liquid the anion plays a larger role in dictating the ultimate solubility of the silk. The dissolution of the silk in the ionic liquid is confirmed using wide-angle X-ray scattering. The dissolved silk is also processed into 100 mum-thick, two-dimensional films, and the structure of these films is examined. The rinse solvent, acetonitrile or methanol, has a profound impact on both the topography of the films and the secondary structure of the silk protein. The image depicts a silkworm cocoon dissolved in 1-butyl-3-methylimidazolium chloride and then regenerated as a film with birefringence. PMID- 15521744 TI - Identification and characterization of monoanionic tripodal tetradentate ligand complexes of copper(I) and copper(II) involved in halogen atom transfer reactions. AB - The copper(I) complex of bis-(2-(2-pyridyl)-ethyl)-(2-(N-p-toluenesulfonamido) ethyl)amine (PETAEA), a monoanionic, tripodal tetradentate ligand, was prepared, characterized, and shown to be an effective catalyst for atom transfer radical polymerization (ATRP). A model atom transfer reaction of Cu(PETAEA) with 1 phenylethyl bromide and TEMPO radical trapping agent was studied. The copper(II) complex formed in this reaction was identified by comparison of its spectroscopic data with that of Cu(PETAEA)Br prepared by an independent synthesis. Kinetic and spectroscopic data indicated that the reaction mechanism involved simple atom transfer from the alkyl halide to the Cu(PETAEA) to form the Cu(PETAEA)Br, and no other intermediates were involved. The solid-state structures of the copper(I) and (II) complexes appeared to be maintained in solution, so this system is an atom transfer reaction in which all of the reactive species are identified and characterized. PMID- 15521745 TI - Specific synthesis of 1,2- and 1,3-dialkylidenecycloheptanes by [3+2+2] cyclization of alkenyl Fischer carbene complexes and allenes. AB - The 1,2- and 1,3-dialkylidenecycloheptane rings are specifically assembled from chromium alkenyl Fischer carbene complexes and allenes via [3+2+2] cyclization reactions. The former cycloadducts are obtained when the cyclization is performed in the presence of 1 equiv of [Ni(cod)2], while the [Rh(cod)Cl]2-catalyzed cyclization leads to the latter cycloadducts. PMID- 15521746 TI - Vibration-rotation emission spectra of gaseous ZnH2 and ZnD2. AB - Gaseous ZnH2 and ZnD2 have been discovered in an emission source that combines an electrical discharge with a high-temperature furnace. High-resolution infrared emission spectra of ZnH2 and ZnD2 have been recorded with a Fourier transform spectrometer, and the antisymmetric stretching fundamental bands of 64ZnH2 and 64ZnD2 were detected near 1889.4 and 1371.6 cm-1, respectively. Rotational analysis of the bands yielded r0 bond distances of 1.535 271(1) and 1.531 836(9) A for linear 64ZnH2 and 64ZnD2, respectively. PMID- 15521747 TI - Total synthesis of (+/-)-jiadifenin, a non-peptidyl neurotrophic modulator. AB - We report the first total synthesis of jiadifenin (1), the establishment of a modality for its biological evaluation, and the discovery of apparently more potent neurotrophic activity in fully synthetic compound 17, an intermediate en route to 1. PMID- 15521748 TI - Highly stereo- and regioselective ni-catalyzed homoallylation of aldimines with conjugated dienes promoted by diethylzinc. AB - Under the nickel catalysis, in one pot, diethylzinc serves as a reducing agent to connect the three components of 1,3-dienes, alkyl or aryl aldehydes, and anilines to furnish 3-substituted 3,5-syn-5-(arylamino)pent-1-enes 1, the homoallylation products of aldimines with dienes, in excellent yields and with high regio- and stereoselectivity. PMID- 15521749 TI - Drug binding revealed by tandem mass spectrometry of a protein-micelle complex. AB - The protein-micelle complex formed between the protein EmrE and the lipid dodecylmaltoside has been examined by mass spectrometry. The results show that despite the unfavorable hydrophobic environment in the mass spectrometer it is possible to preserve protein submicelle complexes in the gas phase. The peaks assigned to the submicelle complexes are broad in nature and consistent with a heterogeneous distribution of lipid molecules attached to the protein complex. As such, the spectrum cannot be interpreted. To simplify this complexity we used a tandem mass spectrometry procedure in which discrete m/z values are isolated from the peak and subjected to collision-induced dissociation. These spectra reveal clusters of DDM molecules as well as sequential release of TPP+ and EmrE from the complex as the collision cell voltage is raised. Taken together, the results provide direct evidence for drug binding within a relevant gas-phase protein micelle complex. PMID- 15521750 TI - Energy transfer rates and pathways of single donor chromophores in a multichromophoric dendrimer built around a central acceptor core. AB - An artificial light-harvesting dendrimer showing highly efficient electronic excitation energy transfer from four peripheral donors to one central acceptor has been investigated by single-molecule spectroscopy at low temperatures. Confocal imaging in combination with frequency selective excitation spectroscopy gives direct access to energy transfer rates of individual donors and allows the determination of energy transfer pathways within a single multichromophoric aggregate. PMID- 15521751 TI - Controlling photochemistry with distinct hydrophobic nanoenvironments. AB - A combination of hydrophobic forces and guest templation drive the assembly of cavitands into molecular capsules. Encapsulated guests such as dibenzyl ketones reside in an essentially dry environment, and upon irradiation, undergo rearrangement processes that are templated by the shape of the 1 nm x 2 nm cavity. PMID- 15521752 TI - Unidirectionally aligned copper hydroxide crystalline nanorods from two dimensional copper hydroxy nitrate. AB - We were able to synthesize unidirectionally aligned copper hydroxide nanorods from two-dimensional copper hydroxide nitrate by anion exchange reaction using NaOH without any membrane or template. The structural conversion from Cu2(OH)3NO3 to Cu(OH)2 was confirmed by XRD and FT-IR data. The synthesized copper hydroxide nanorods showed unidirectionally aligned and tightly bound arrays on a hexagonal plate, leading to giant nanorod bundles. PMID- 15521753 TI - N-heterocyclic carbene-catalyzed generation of homoenolates: gamma-butyrolactones by direct annulations of enals and aldehydes. AB - N-Heterocyclic carbenes, prepared in situ from diarylimidazolium salts, serve as highly effective catalysts for the generation of reactive homoenolates from alpha,beta-unsaturated aldehydes. The catalyst-bound homoenolate reacts with electrophilic aldehydes leading, via the key intermediacy of an activated carboxylate, to gamma-butyrolactones in good yields and stereoselectivities. Importantly, this process demonstrates an unprecedented reaction mode for the generation of nucleophilic carbanions with a multifunctional organocatalyst under exceptionally mild and convenient reaction conditions. PMID- 15521754 TI - Crystal orientation-ordered ZnS nanowire bundles. AB - We report a novel approach for growing aligned and orientation-ordered ZnS nanowires. Our method relies on a buffer layer of CdSe grown on a Si(111) substrate, on which ZnS nanowires are grown. The growth process of the nanowire bundles is presented. The technique demonstrated could be an effective pathway for growing patterned, aligned, size-controlled, and orientation-ordered ZnS nanowires. PMID- 15521755 TI - Total synthesis of brevetoxin-B. AB - Brevetoxin-B (BTX-B), produced by the red tide organism, Gymnodium breve Davis, is the first member of marine polycyclic ethers to be structurally elucidated and one of the most potent neurotoxins. The structural feature is a trans-fused polycyclic ether ring system with 23 stereocenters. Its unique, complex structure and potent biological activity have attracted the attention of synthetic organic chemists. Total synthesis of BTX-B has been accomplished via the coupling of the ABCDEFG and IJK-ring segments, each ether ring of which was stereoselectively and efficiently constructed on the basis of SmI2-induced intramolecular cyclization, 6-endo-cyclization of hydroxy epoxide, ring-closing olefin metathesis, and SmI2 induced intramolecular Reformatsky-type reaction. Several kinds of double reactions at the left and right sides were efficiently used through the synthesis. PMID- 15521756 TI - Mass spectrometry assisted assignment of NMR resonances in 15N labeled proteins. AB - Application of nuclear magnetic resonance (NMR) methods for the structural characterization to larger and more complex protein systems can be facilitated through the development of new methods for resonance assignment. Here, a novel approach that relies on integration of NMR and mass spectrometry (MS) methods is explored. The approach relies on the fact that both NMR and MS are able to monitor rates of exchange of amide protons for water deuterons. Correlating the rates can connect cross-peak positions from NMR data with fragment masses from MS data to support sequential assignment. The example provided is for a small model protein, ubiquitin, but the potential for application to large, more difficult to express proteins is clear. PMID- 15521757 TI - Deciphering the origins of observed heat capacity changes for aminoglycoside binding to prokaryotic and eukaryotic ribosomal RNA a-sites: a calorimetric, computational, and osmotic stress study. AB - Isothermal titration calorimetry (ITC), computational, and osmotic stress techniques have been used to characterize the changes in heat capacity, solvent accessible surface, and hydration that accompany the binding of the aminoglycoside paromomycin to both prokaryotic and eukaryotic rRNA A-site model oligonucleotides. Regarded as a whole, the results of these studies suggest that the intrinsic heat capacity change (DeltaC(p)) for the binding of paromomycin to each rRNA A-site is near zero, with the negative DeltaC(p) observed for the binding of the drug to the prokaryotic rRNA A-site being dictated by the coupled destacking of the adenine residues at positions 1492 and 1493. In this connection, DeltaC(p) provides a useful calorimetric signature for assessing the relative impacts of novel and existing A-site targeting ligands on rRNA conformation, which, in turn, should provide a useful analytical tool for facilitating the drug design process, since aminoglycoside-induced destacking of A1492 and A1493 is thought to be a determining factor in the mistranslational and antimicrobial activities of the drugs. PMID- 15521758 TI - Spectroscopic identification of different types of copper centers generated in synthetic four-helix bundle proteins. AB - Using a combined rational-combinatorial approach, stable copper binding sites were implemented in template-assembled synthetic four-helix bundle proteins constructed by three different helices with only 16 amino acid residues. These peptides include two histidines and one cysteine at positions appropriate for coordinating a copper ion. Sequence variations of the helices were made in the second coordination shell or even more remote from the copper binding site (i) to increase the overall stability of the metalloproteins and (ii) to fine-tune the structure and properties of the copper center. As a result, ca. 90% of the 180 proteins that were synthesized were capable to bind copper with a substantially higher specificity than those obtained in the first design cycle (Schnepf, R.; Horth, P.; Bill, E.; Wieghardt, K.; Hildebrandt, P.; Haehnel, W. J. Am. Chem. Soc. 2001, 123, 2186-2195). Furthermore, the stabilities of the copper protein complexes were increased by up to 2 orders of magnitude and thus allowed a UV-vis absorption, resonance Raman, electron paramagnetic resonance, and (magnetic) circular dichroism spectroscopic identification and characterization of three different types of copper binding sites. It could be shown that particularly steric perturbations in the vicinity of the His(2)Cys ligand set control the formation of either a tetragonal (type II) or a tetrahedral (type I) copper binding site. With the introduction of two methionine residues above the histidine ligands, a mixed-valent dinuclear copper binding site was generated with spectroscopic properties that are very similar to those of Cu(A) sites in natural proteins. The results of the present study demonstrate for the first time that structurally different metal binding sites can be formed and stabilized in four-helix bundle proteins. PMID- 15521759 TI - Interactions of alkyltin salts with biological dithiols: dealkylation and induction of a regular beta-turn structure in peptides. AB - Organotin compounds specifically target vicinal dithiols, thereby inhibiting the function of essential enzymes. Here, we present the NMR binding studies of trimethyltin (TMT) and dimethyltin (DMT) chlorides with a linear peptide (ILGCWCYLR) derived from the membrane protein stannin (SNN). We show that this peptide is able to dealkylate TMT and bind DMT, adopting a stable type-I beta turn conformation. Both the NMR data and the calculated structures indicate that the two cysteines coordinate the tin atom in a distorted tetrahedral geometry. The molecular geometries and tin coordination state were confirmed using density functional theory (DFT). In addition, NMR spectral parameters back calculated from the DFT minimized structure compared well with experimental data. These results in conjunction with studies on peptide variants (i.e., C4S, C6S, and Y7F) demonstrate unequivocally the key role of biological dithiols in both the dealkylation and binding of organotin compounds. This peptide serves as a model system for alkyltin-protein interactions and gives new insights into the biological fate of alkyltin compounds. PMID- 15521760 TI - Artificial metalloenzymes: (strept)avidin as host for enantioselective hydrogenation by achiral biotinylated rhodium-diphosphine complexes. AB - We report on the generation of artificial metalloenzymes based on the noncovalent incorporation of biotinylated rhodium-diphosphine complexes in (strept)avidin as host proteins. A chemogenetic optimization procedure allows one to optimize the enantioselectivity for the reduction of acetamidoacrylic acid (up to 96% ee (R) in streptavidin S112G and up to 80% ee (S) in WT avidin). The association constant between a prototypical cationic biotinylated rhodium-diphosphine catalyst precursor and the host proteins was determined at neutral pH: log K(a) = 7.7 for avidin (pI = 10.4) and log K(a) = 7.1 for streptavidin (pI = 6.4). It is shown that the optimal operating conditions for the enantioselective reduction are 5 bar at 30 degrees C with a 1% catalyst loading. PMID- 15521761 TI - Optimization of interstrand hydrophobic packing interactions within unnatural DNA base pairs. AB - As part of an effort to expand the genetic alphabet, we have evaluated a large number of predominantly hydrophobic unnatural base pairs. We now report the synthesis and stability of unnatural base pairs formed between simple phenyl rings modified at different positions with methyl groups. Surprisingly, several of the unnatural base pairs are virtually as stable as a natural base pair in the same sequence context. The results show that neither hydrogen-bonding nor large aromatic surface area are required for base pair stability within duplex DNA and that interstrand interactions between small aromatic rings may be optimized for both stability and selectivity. These smaller nucleobases are not expected to induce the distortions in duplex DNA or at the primer terminus that seem to limit replication of larger unnatural base pairs, and they therefore represent a promising approach to the expansion of the genetic alphabet. PMID- 15521762 TI - The structure of Ba@C74. AB - The title compound has been produced by using the radio frequency (RF) method. Barium and carbon were evaporated simultaneously under dynamic flow of helium at different temperatures. About 0.5 mg of pure Ba@C(74) was isolated via a three step high-pressure liquid chromatography separation. For the first time, the structure of a monometallofullerene has been analyzed by means of single-crystal synchrotron diffraction on microcrystals of Ba@C(74).Co(OEP).2C(6)H(6) (Co(II)(OEP): cobalt(II) octaethylporphyrin) at 100 K. The monometallofullerene exhibits a high degree of localization of the endohedral metal ion, with just two split positions for Ba and two orientations of the C(74)-cage. The barium atom is localized inside the C(74)-cage and displaced off-center, toward the Co(OEP) molecule (d approximately 127 pm). The shortest Ba-C distance is 265 pm. The Co(OEP) molecules form dimers in which the coordination of the cobalt is (4 + 1). Due to the all-syn conformation of the ethyl groups, each Co(OEP) molecule of the dimer coordinates one C(74)-fullerene. The units (Ba@C(74))[Co(OEP)](2)(Ba@C(74)) are arranged in a distorted primitive hexagonal packing. The free space between these complex units is filled by benzene molecules of crystallization. The Ba L(III) XANES spectrum of a thin film sample of Ba@C(74) exhibits a pronounced double maximum structure at about E = 5275 eV. The comparison of the shape resonances of the experimental data with simulated XANES spectra, based on different exo- and endohedral structure models, confirm that the Ba atom is located inside the C(74)-cage (D(3)(h)()) in an off-center position. The Ba atom is shifted by about 130-150 pm from the geometric center of the C(74)-cage. This is in good agreement with quantum chemical results. Thus, despite the disorder still present, a consistent and conclusive structure model for the title compound has been derived by employing a combination of X-ray diffraction, XANES spectroscopy, and quantum chemical calculations. PMID- 15521763 TI - Developing photoactive affinity probes for proteomic profiling: hydroxamate-based probes for metalloproteases. AB - The denaturing aspect of current activity-based protein profiling strategies limits the classes of chemical probes to those which irreversibly and covalently modify their targeting enzymes. Herein, we present a complimentary, affinity based labeling approach to profile enzymes which do not possess covalently bound substrate intermediates. Using a variety of enzymes belonging to the class of metalloproteases, the feasibility of the approach was successfully demonstrated in several proof-of-concept experiments. The design template of affinity-based probes targeting metalloproteases consists of a peptidyl hydroxamate zinc-binding group (ZBG), a fluorescent reporter tag, and a photolabile diazirine group. Photolysis of the photolabile unit in the probe effectively generates a covalent, irreversible linkage between the probe and the target enzyme, rendering the enzyme distinguishable from unlabeled proteins upon separation on a SDS-PAGE gel. A variety of labeling studies were carried out to confirm that the affinity-based approach selectively labeled metalloproteases in the presence of a large excess of other proteins and that the success of the labeling reaction depends intimately upon the catalytic activity of the enzyme. Addition of competitive inhibitors proportionally diminished the extent of enzyme labeling, making the approach useful for potential in situ screening of metalloprotease inhibitors. Using different probes with varying P(1) amino acids, we were able to generate unique "fingerprint" profiles of enzymes which may be used to determine their substrate specificities. Finally, by testing against a panel of yeast metalloproteases, we demonstrated that the affinity-based approach may be used for the large-scale profiling of metalloproteases in future proteomic experiments. PMID- 15521764 TI - Strand orientation by steric matching: a designed antiparallel coiled-coil trimer. AB - The design of an antiparallel coiled-coil 1:1:1 heterotrimer is described. Control of strand orientation results from proper alignment of sterically matched hydrophobic core side chains. Matched core layers position one cyclohexylalanine side chain against two alanine ones. Substitution of three consecutive heptad a positions with all permutations of two alanines and one cyclohexylalanine (AAX, AXA, AAX, where A = alanine, X = cyclohexylalanine) affords a parallel 1:1:1 heterotrimer, as previously reported. Here, we report that moving the substitution sites in one strand to d rather than a positions affords a new peptide that can form an antiparallel complex with the other original components. The new assembly is characterized by circular dichroism spectroscopy ([theta;](222) = -30 317 deg cm(2) dmol(-1), T(m) = 77 degrees C, DeltaG(unf) = 17.1 kcal/mol), and its stoichiometry and aggregation number are confirmed by nickel tag affinity analysis and analytical ultracentrifugation. Disulfide exchange data support the preference for an antiparallel arrangement. Examination of the functionally identical parallel complex demonstrates that the antiparallel structure is comparably stable, as confirmed by a direct competition assay that established an equilibrium 55:45 ratio of each assembly. PMID- 15521765 TI - Synthesis, self-assembly, and characterization of supramolecular polymers from electroactive dendron rodcoil molecules. AB - We report here the synthesis and self-assembly of a series of three molecules with dendron rodcoil architecture that contain conjugated segments of oligo(thiophene), oligo(phenylene-vinylene), and oligo(phenylene). Despite their structural differences, all three molecules yield similar self-assembled structures. Electron and atomic force microscopy reveals the self-assembly of the molecules into high aspect ratio ribbon-like nanostructures which at low concentrations induce gelation in nonpolar solvent. Self-assembly results in a blue-shifted absorption spectrum and a red-shifted, quenched fluorescence spectrum, indicating aggregation of the conjugated segments within the ribbon like structures. The assembly of these molecules into one-dimensional nanostructures is a route to pi-pi stacked supramolecular polymers for organic electronic functions. In the oligo(thiophene) derivative, self-assembly leads to a 3 orders of magnitude increase in the conductivity of iodine-doped films due to self-assembly. We also found that electric field alignment of these supramolecular assemblies can be used to create arrays of self-assembled nanowires on a device substrate. PMID- 15521766 TI - Simulated evolution of emergent chiral structures in polyalanine. AB - The relationship between monomer chirality and polymer structure has been studied using both theoretical and experimental methods. Atomistic models, such as the ones employed in computational protein folding and design, can be used to study the relationship between monomer chirality and the properties of polypeptides. Using a simulated evolution approach that combines side-chain epimerization with backbone flexibility, we recapitulate the relationship between basic forces that drive secondary structure formation and sequence homochirality. Additionally, we find heterochiral motifs including a C-terminal helix capping interaction and stable helix-reversals that result in bent helix structures. Our studies show that simulated evolution of chirality with backbone flexibility can be a powerful tool in the design of novel heteropolymers with tuned stereochemical properties. PMID- 15521767 TI - Bimolecular homolytic substitution (S(H)2) reactions with hydrogen atoms. time resolved electron spin resonance detection in the pulse radiolysis of alpha (methylthio)acetamide. AB - Pulse radiolysis of aqueous solutions of alpha-(methylthio)acetamide produced unexpectedly large quantities of acetamide radicals that were identified by time resolved electron spin resonance (TRESR) spectroscopy. The pH dependence of the TRESR-measured radical yields, results from selective scavenging reactions, and density functional theory predictions of the reaction thermochemistry prove that bimolecular homolytic substitution, S(H)2, of the acetamide radical fragment by a H atom is the most likely formation pathway. PMID- 15521768 TI - Palladium- and copper-catalyzed synthesis of medium- and large-sized ring-fused dihydroazaphenanthrenes and 1,4-benzodiazepine-2,5-diones. control of reaction pathway by metal-switching. AB - Methods for the synthesis of dihydroazaphenanthrene fused to macrocycles (2) and medium-ring heterocycles (4), as well as 1,4-benzodiazepine-2,5-diones (5), are developed. A distinctly different catalytic property of palladium and copper catalysts was uncovered that leads to the development of a divergent synthesis of two different heterocyclic scaffolds from the same starting materials, simply by metal-switching. Thus, starting from linear amide 3, palladium acetate triggers a domino intramolecular N-arylation/C-H activation/aryl-aryl bond-forming process to provide 4, while copper iodide promotes only the intramolecular N-arylation reaction leading to 5. In combination with the Ugi multicomponent reaction (Ugi 4CR) for the preparation of the linear amides, a two-step synthesis of either the 5,6-dihydro-8H-5,7a-diazacyclohepta[jk]phenanthrene-4,7-dione (4) or 1,4 benzodiazepine-2,5-diones (5), by appropriate choice of metal catalyst, is subsequently developed from very simple starting materials. PMID- 15521769 TI - Detection of transient radical cations in electron transfer-initiated Diels-Alder reactions by electrospray ionization mass spectrometry. AB - The coupling of a simple microreactor to an atmospheric pressure ion source, such as electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI), allows the investigation of reactions in solution by mass spectrometry. The tris(p-bromophenyl)aminium hexachloroantimonate (1(*)(+)SbCl(6)(-))-initiated reactions of phenylvinylsulfide (2) and cyclopentadiene (3) and of trans-anethole (5) and isoprene (6) and the dimerization of 1,3-cyclohexadiene (8) to give the respective Diels-Alder products were studied. These preparatively interesting reactions proceed as radical cation chain reactions via the transient radical cations of the respective dienophiles and of the respective Diels-Alder addition products. These radical cations could be detected directly and characterized unambiguously in the reacting solution by ESI-MS-MS. The identity was confirmed by comparison with MS-MS spectra of the authentic radical cations obtained by APCI-MS and by CID experiments of the corresponding molecular ions generated by EI-MS. In addition, substrates and products could be monitored easily in the reacting solution by APCI-MS. PMID- 15521770 TI - Spirocyclic zwitterionic lambda5Si-silicates with two bidentate ligands derived from alpha-amino acids or alpha-hydroxycarboxylic acids: synthesis, structure, and stereodynamics. AB - A series of zwitterionic lambda(5)Si-silicates with a (2,2,6,6 tetramethylpiperidinio)methyl group and two identical bidentate ligands derived from glycine, (S)-alanine, (S)-phenylalanine, glycolic acid, (S)-lactic acid, (S) 3-phenyllactic acid, or (S)-mandelic acid were synthesized and structurally characterized (solution and solid-state NMR spectroscopy; single-crystal X-ray diffraction). The chiral lambda(5)Si-silicates with ligands derived from optically active alpha-amino acids or alpha-hydroxycarboxylic acids were isolated as enantiomerically and diastereomerically pure compounds that undergo a (Lambda)/(Delta)-epimerization in solution. PMID- 15521771 TI - Evidence for through-space electron transfer in the distance dependence of normal and inverted electron transfer in oligoproline arrays. AB - Four new helical oligoproline assemblies containing 16, 17, 18, and 19 proline residues and ordered arrays of a Ru(II)-bipyridyl chromophore and a phenothiazine electron-transfer donor have been synthesized in a modular fashion by solid-phase peptide synthesis. These arrays are illustrated and abbreviated as CH(3)CO-Pro(6) Pra(PTZ)-Pro(n)()-Pra(Ru(II)b(2)m)(2+)-Pro(6)-NH(2), where PTZ is 3-(10H phenothiazine-10)propanoyl and (Ru(II)b'(2)m)(2+) is bis(4,4'-diethylamide-2,2' bipyridine)(4-methyl,4'-carboxylate,2,2'-bipyridine)ruthenium(II) dication with n = 2 (2), 3 (3), 4 (4), and 5 (5). They contain PTZ as an electron-transfer donor and (Ru(II)b'(2)m)(2+) as a metal-to-ligand charge transfer (MLCT) light absorber and are separated by proline-to-proline through-space distances ranging from 0 (n = 2) to 12.9 A (n = 5) relative to the n = 2 case. They exist in the proline-II helix form in water, as shown by circular dichroism measurements. Following laser flash Ru(II) --> b'(2)m MLCT excitation at 460 nm in water, excited-state PTZ --> Ru(2+) quenching (k(2)) occurs by reductive electron transfer, followed by Ru(+) -> PTZ(+) back electron transfer (k(3)), as shown by transient absorption and emission measurements in water at 25 degrees C. Quenching with DeltaG degrees = 0.1 eV is an activated process, while back electron transfer occurs in the inverted region, DeltaG degrees = -1.8 eV, and is activationless, as shown by temperature dependence measurements. Coincidentally, both reactions have comparable distance dependences, with k(2)( )()varying from = 1.9 x 10(9) (n = 2) to 2.2 x 10(6) s(-)(1) (n = 4) and k(3) from approximately 2.0 x 10(9) (n = 2) to 2.2 x 10(6) s(-)(1) (n = 4). For both series there is a rate constant enhancement of approximately 10 for n = 5 compared to n = 4 and a linear decrease in ln k with the through-space separation distance, pointing to a significant and probably dominant through-space component to intrahelical electron transfer. PMID- 15521772 TI - Dihydrogen and methane elimination from adducts formed by the interaction of carbenium and silylium cations with nucleophiles. AB - Stationary points for reactions R'R' 'HX(+) + YH --> [R'R' 'X-Y](+) + H(2) (I) and R'(CH(3))HX(+) + YH -->[R'HX-Y](+) + CH(4) (II) (R', R' ' = CH(3), H; X = C, Si; Y = CH(3)O, (CH(3))(2)N, and C(6)H(5)) are located and optimized by the B3LYP/aug-cc-pVDZ method. A similar mechanism was found to be operative for both types of reactions with X = C and X = Si. Formation of the intermediate (adduct) results in the transfer of electron density from the electron-rich bases to the X atoms and in the growth of a positive charge on a hydrogen atom attached to Y. This mobile proton may shift from Y to X, and the relative energies of transition states for elimination reactions (Delta) depend on the ability of the X atom to retain this proton. Therefore, Deltagrows on going from Si to C and with increasing numbers of methyl substituents. For X = C, the Deltavalue for both reactions correlates well with the population of the valence orbitals of X in a wide range from -44 kcal/mol (methyl cation/benzene) to 31 kcal/mol (isopropyl cation/methanol). For X = Si this range is more narrow (from -19 to -5.0 kcal/mol), but all Delta values are negative with the exclusion of silylium ion/benzene systems, adducts of which are pi- rather than sigma-complexes. The energy minima for product complexes for H(2) elimination are very shallow, and several are dissociative. However, complexes with methane which exhibit bonding between X and the methane hydrogen are substantially stronger, especially for systems with X = Si. The latter association energy may reach 8 kcal/mol (Si...H distance is 2 A). PMID- 15521773 TI - A spectroscopic investigation of carbon-carbon bond formation by methylene insertion on a Ag(111) surface: mechanism and kinetics. AB - Using reflection-absorption infrared spectroscopy (RAIRS) and temperature programmed reaction spectroscopy (TPRS), we have investigated the cross-coupling reaction between CH(2)(a) and CF(3)(a) on a Ag(111) surface. CH(2)(a) and CF(3)(a) are generated by thermal decomposition of adsorbed CH(2)I(2) and CF(3)I. RAIRS results unambiguously demonstrate that CH(2)(a) inserts into the Ag-CF(3) bond to produce adsorbed CF(3)CH(2)(a), which upon heating selectively undergoes beta-fluorine elimination to form CH(2)=CF(2). Increasing the CH(2)(a) and CF(3)(a) coverage leads to the sequential insertion of CH(2)(a) into Ag-CF(3), as evidenced by CH(2)CH(2)CF(3)(a) formation detected with RAIRS. Prior to the insertion reaction, the evidence favors islanding of fragments. The methylene insertion reaction is so facile that it occurs at cryogenic temperatures (120 K). Time-resolved RAIRS (TR-RAIRS) results at selected temperatures reveal an activation energy of 5.8 kJ/mol. Our results provide, for the first time, direct spectroscopic information about the mechanism and kinetics of the methylene insertion reaction. PMID- 15521774 TI - Structure, reactivity, and density functional theory analysis of the six-electron reductant, [(C5Me5)2U]2(mu-eta6:eta6-C6H6), synthesized via a new mode of (C5Me5)3M reactivity. AB - The sterically crowded (C(5)Me(5))(3)U complex reacts with KC(8) or K/(18-crown 6) in benzene to form [(C(5)Me(5))(2)U](2)(mu-eta(6):eta(6)-C(6)H(6)), 1, and KC(5)Me(5). These reactions suggested that (C(5)Me(5))(3)U could be susceptible to (C(5)Me(5))(1-) substitution by benzene anions via ionic salt metathesis. To test this idea in the synthesis of a more conventional product, (C(5)Me(5))(3)U was treated with KN(SiMe(3))(2) to form (C(5)Me(5))(2)U[N(SiMe(3))(2)] and KC(5)Me(5). 1 has long U-C(C(5)Me(5)) bond distances comparable to (C(5)Me(5))(3)U, and it too is susceptible to (C(5)Me(5))(1-) substitution via ionic metathesis: 1 reacts with KN(SiMe(3))(2) to make its amide-substituted analogue [[(Me(3)Si)(2)N](C(5)Me(5))U](2)(mu-eta(6):eta(6)-C(6)H(6)), 2. Complexes 1 and 2 have nonplanar C(6)H(6)-derived ligands sandwiched between the two uranium ions. 1 and 2 were examined by reactivity studies, electronic absorption spectroscopy, and density functional theory calculations. [(C(5)Me(5))(2)U](2)(mu-eta(6):eta(6)-C(6)H(6)) functions as a six-electron reductant in its reaction with 3 equiv of cyclooctatetraene to form [(C(5)Me(5))(C(8)H(8))U](2)(mu-eta(3):eta(3)-C(8)H(8)), (C(5)Me(5))(2), and benzene. This multielectron transformation can be formally attributed to three different sources: two electrons from two U(III) centers, two electrons from sterically induced reduction by two (C(5)Me(5))(1-) ligands, and two electrons from a bridging (C(6)H(6))(2-) moiety. PMID- 15521775 TI - Envisaging the physicochemical processes during the preparation of supported catalysts: Raman microscopy on the impregnation of Mo onto Al2O3 extrudates. AB - Raman microscopy has been applied to study the preparation of shaped Mo/Al(2)O(3) catalysts. The speciation of different Mo complexes over gamma-Al(2)O(3) support bodies was followed in time after pore volume impregnation with aqueous solutions containing different Mo complexes. The addition of NO(3-) to the impregnation solutions allows for a quantitative Raman analysis of the distribution of different complexes over the catalyst bodies as this ion can be used as an internal standard. After impregnation with an acidic ammonium heptamolybdate (AHM) solution, the strong interaction between Mo(7)O(24)(6-) and Al(2)O(3) results in slow transport of this complex through the support and extensive formation of Al(OH)(6)Mo(6)O(18)(3-) near the outer surface of the support bodies. This may be prevented by decreasing the interaction between Mo and Al(2)O(3). In this way, transport is facilitated and a homogeneous distribution of Mo is obtained on a reasonable time scale. A decrease in interaction between Mo and Al(2)O(3) can be achieved by using alkaline impregnation solutions or by the addition of complexing agents, such as citrate and phosphate, to the impregnation solution. In general, time-resolved in situ Raman microscopy can be a valuable tool to study the physicochemical processes during the preparation of supported catalysts. PMID- 15521776 TI - Noncovalent anchoring of homogeneous catalysts to silica supports with well defined binding sites. AB - The efficient reversible functionalization of silica with catalytic sites using noncovalent interactions is described. We prepared silica materials with well defined binding sites that selectively bind guest molecules that are equipped with the complementary binding motif, with the interaction between the two components being based on either hydrogen bonds or metal-ligand interactions. Several phosphine ligands functionalized with glycine-urea groups, required for hydrogen bond formation to the complementary host on the silica, have been prepared. The resulting noncovalently immobilized complexes have been used as a ligand system in the Pd-catalyzed allylic substitution and Rh-catalyzed hydroformylation of 1-octene. The supramolecular interaction between the transition-metal catalyst and the binding site located at the support is sufficiently strong to enable efficient catalyst recycling. In addition, the nature of the support facilitates the de- and refunctionalization of support, allowing the recycling of both homogeneous catalysts and the functionalized support. A rhodium catalyst based on a functionalized xantphos ligand was used in the hydroformylation of 1-octene in 11 consecutive reactions without showing catalyst deterioration or metal leaching. PMID- 15521777 TI - Concomitant length and diameter separation of single-walled carbon nanotubes. AB - Gel electrophoresis and column chromatography conducted on individually dispersed, ultrasonicated single-walled carbon nanotubes yield simultaneous separation by tube length and diameter. Electroelution after electrophoresis is shown to produce highly resolved fractions of nanotubes with average lengths between 92 and 435 nm. Separation by diameter is concomitant with length fractionation, and nanotubes that have been cut shortest also possess the greatest relative enrichments of large-diameter species. Longer sonication time causes increased electrophoretic mobility in the gels; thus, ultrasonic processing determines the degree of both length and diameter separation of the nanotubes. The relative quantum yield decreases nonlinearly as the nanotube length becomes shorter. These techniques constitute a preparative, scalable method for separating nanotubes by two important attributes required for electronic and sensor applications. PMID- 15521778 TI - Expanding dinitrogen reduction chemistry to trivalent lanthanides via the LnZ3/alkali metal reduction system: evaluation of the generality of forming Ln2(mu-eta2:eta2-N2) complexes via LnZ3/K. AB - The Ln[N(SiMe(3))(2)](3)/K dinitrogen reduction system, which mimicks the reactions of the highly reducing divalent ions Tm(II), Dy(II), and Nd(II), has been explored with the entire lanthanide series and uranium to examine its generality and to correlate the observed reactivity with accessibility of divalent oxidation states. The Ln[N(SiMe(3))(2)](3)/K reduction of dinitrogen provides access from readily available starting materials to the formerly rare class of M(2)(mu-eta(2):eta(2)-N(2)) complexes, [[(Me(3)Si)(2)N](2)(THF)Ln](2)(mu eta(2):eta(2)-N(2)), 1, that had previously been made only from TmI(2), DyI(2), and NdI(2) in the presence of KN(SiMe(3))(2). This LnZ(3)/alkali metal reduction system provides crystallographically characterizable examples of 1 for Nd, Gd, Tb, Dy, Ho, Er, Y, Tm, and Lu. Sodium can be used as the alkali metal as well as potassium. These compounds have NN distances in the 1.258(3) to 1.318(5) A range consistent with formation of an (N=N)(2)(-) moiety. Isolation of 1 with this selection of metals demonstrates that the Ln[N(SiMe(3))(2)](3)/alkali metal reaction can mimic divalent lanthanide reduction chemistry with metals that have calculated Ln(III)/Ln(II) reduction potentials ranging from -2.3 to -3.9 V vs NHE. In the case of Ln = Sm, which has an analogous Ln(III)/Ln(II) potential of 1.55 V, reduction to the stable divalent tris(amide) complex, K[Sm[N(SiMe(3))(2)](3)], is observed instead of dinitrogen reduction. When the metal is La, Ce, Pr, or U, the first crystallographically characterized examples of the tetrakis[bis(trimethylsilyl)amide] anions, [M[N(SiMe(3))(2)](4)](-), are isolated as THF-solvated potassium or sodium salts. The implications of the LnZ(3)/alkali metal reduction chemistry on the mechanism of dinitrogen reduction and on reductive lanthanide chemistry in general are discussed. PMID- 15521779 TI - Magnetic, electronic, and structural characterization of nonstoichiometric iron oxides at the nanoscale. AB - We have investigated the structural, magnetic, and electronic properties of nonstoichiometric iron oxide nanocrystals prepared by decomposition of iron(II) and iron(0) precursors in the presence of organic solvents and capping groups. The highly uniform, crystalline, and monodisperse nanocrystals that were produced enabled a full structural and compositional survey by electron microscopy and X ray diffraction. The complex and metastable behavior of nonstoichiometric iron oxide (wustite) at the nanoscale was studied by a combination of Mossbauer spectroscopy and magnetic characterization. Deposition from hydrocarbon solvents with subsequent self-assembly of iron oxide nanocrystals into superlattices allowed the preparation of continuous thin films suitable for electronic transport measurements. PMID- 15521780 TI - Cation-pi interactions: structures and energetics of complexation of Na+ and K+ with the aromatic amino acids, phenylalanine, tyrosine, and tryptophan. AB - Threshold collision-induced dissociation of M(+)(AAA) with Xe is studied using guided ion beam tandem mass spectrometry. M(+) include the alkali metal ions Na(+) and K(+). The three aromatic amino acids are examined, AAA = phenylalanine, tyrosine, or tryptophan. In all cases, endothermic loss of the intact aromatic amino acid is the dominant reaction pathway. The threshold regions of the cross sections are interpreted to extract 0 and 298 K bond dissociation energies for the M(+)-AAA complexes after accounting for the effects of multiple ion-neutral collisions, internal energy of the reactant ions, and dissociation lifetimes. Density functional theory calculations at the B3LYP/6-31G level of theory are used to determine the structures of the neutral aromatic amino acids and their complexes to Na(+) and K(+) and to provide molecular constants required for the thermochemical analysis of the experimental data. Theoretical bond dissociation energies are determined from single-point energy calculations at the B3LYP/6 311++G(3df,3pd) level using the B3LYP/6-31G geometries. Good agreement between theory and experiment is found for all systems. The present results are compared to earlier studies of these systems performed via kinetic and equilibrium methods. The present results are also compared to the analogous Na(+) and K(+) complexes to glycine, benzene, phenol, and indole to elucidate the relative contributions that each of the functional components of these aromatic amino acids make to the overall binding in these complexes. PMID- 15521781 TI - On the competition between linear and cyclic isomers in second-row dicarbides. AB - Second-row dicarbides C(2)X (X = Na-Cl) are investigated with quantum mechanical techniques. The cyclic-linear competition in these systems is studied, and the bonding scheme for these compounds is discussed in terms of the topological analysis of the electronic density. C(2)Na, C(2)Mg, C(2)Al, and C(2)Si are found to prefer a C(2)(v)-symmetric arrangement corresponding to a T-shape structure. On the other hand, for C(2)P, C(2)S, and C(2)Cl the linear isomer is predicted to be the ground state. A detailed analysis of the variation of the electronic energy and orbital energies with the geometry has been carried out. A simple theoretical model, taking into account the main interactions between the valence orbitals of both fragments, the X atom and the C(2) molecule, allows an interpretation of the main features of these compounds. PMID- 15521782 TI - Interfacial electron-transfer kinetics of ferrocene through oligophenyleneethynylene bridges attached to gold electrodes as constituents of self-assembled monolayers: observation of a nonmonotonic distance dependence. AB - The standard heterogeneous electron-transfer rate constants (k(n)0) between substrate gold electrodes and the ferrocene redox couple attached to the electrode surface by variable lengths of substituted or unsubstituted oligophenyleneethynylene (OPE) bridges as constituents of mixed self-assembled monolayers were measured as a function of temperature. The distance dependences of the unsubstituted OPE standard rate constants and of the preexponential factors (An) obtained from an Arrhenius analysis of the unsubstituted OPE k(n)0 versus temperature data are not monotonic. This surprising result, together with the distance dependence of the substituted OPE preexponential factors, may be assessed in terms of the likely conformational variability of the OPE bridges (as a result of the low intrinsic barrier to rotation of the phenylene rings in these bridges) and the associated sensitivity of the rate of electron transfer (and, hence, the single-molecule conductance which may be estimated using An) through these bridges to the conformation of the bridge. Additionally, the measured standard rate constants were independent of the identity of the diluent component of the mixed monolayer, and using an unsaturated OPE diluent has no effect on the rate of electron transfer through a long-chain alkanethiol bridge. These observations indicate that the diluent does not participate in the electron transfer event. PMID- 15521783 TI - Theoretical studies on the deacylation step of serine protease catalysis in the gas phase, in solution, and in elastase. AB - The deacylation step of serine protease catalysis is studied using DFT and ab initio QM/MM calculations combined with MD/umbrella sampling calculations. Free energies of the entire reaction are calculated in the gas phase, in a continuum solvent, and in the enzyme elastase. The calculations show that a concerted mechanism in the gas phase is replaced by a stepwise mechanism when solvent effects or an acetate ion are added to the reference system, with the tetrahedral intermediate being a shallow minimum on the free energy surface. In the enzyme, the tetrahedral intermediate is a relatively stable species ( approximately 7 kcal/mol lower in energy than the transition state), mainly due to the electrostatic effects of the oxyanion hole and Asp102. It is formed in the first step of the reaction, as a result of a proton transfer from the nucleophilic water to His57 and of an attack of the remaining hydroxyl on the ester carbonyl. This is the rate-determining step of the reaction, which requires approximately 22 kcal/mol for activation, approximately 5 kcal/mol less than the reference reaction in water. In the second stage of the reaction, only small energy barriers are detected to facilitate the proton transfer from His57 to Ser195 and the breakdown of the tetrahedral intermediate. Those are attributed mainly to a movement of Ser195 and to a rotation of the His57 side chain. During the rotation, the imidazolium ion is stabilized by a strong H-bond with Asp102, and the C(epsilon)(1)-H...O H-bond with Ser214 is replaced by one with Thr213, suggesting that a "ring-flip mechanism" is not necessary as a driving force for the reaction. The movements of His57 and Ser195 are highly correlated with rearrangements of the binding site, suggesting that product release may be implicated in the deacylation process. PMID- 15521784 TI - Theoretical study of the guanine --> 6-thioguanine substitution in duplexes, triplexes, and tetraplexes. AB - Molecular dynamics and thermodynamic integration calculations have been carried out on a set of G-rich single-strand, duplex, triplex, and quadruplex DNAs to study the structural and stability changes connected with the guanine --> 6 thioguanine (G --> S) mutation. The presence of 6-thioguanine leads to a shift of the geometry from the B/A intermediate to the pure B-form in duplex DNA. The G - > S mutation does not largely affect the structure of the antiparallel triplex when it is located at the reverse-Hoogsteen position, but leads to a non negligible local distortion in the structure when it is located at the Watson Crick position. The G --> S mutation leads to destabilization of all studied structures: the lowest effect has been observed for the G --> S mutation in the reverse-Hoogsteen strand of the triplex, a medium effect has been observed in the Watson-Crick strand of the triplex and duplex, and the highest influence of the G -->S mutation has been found for the quadruplex structures. PMID- 15521785 TI - Predicting 9Be nuclear magnetic resonance chemical shielding tensors utilizing density functional theory. AB - The structures of a series of beryllium containing complexes have been optimized at the B3LYP/6-31G(d) level and their (9)Be magnetic shielding values have been determined using B3LYP/6-311G+g(2d,p) and the gauge-including atomic orbital (GIAO) method. The calculated chemical shifts are in excellent agreement with experimental values. The performance of a variety of NMR methods (SGO, IGAIM, CSGT) were also examined but were found to be inferior to the GIAO method at the chosen level of theory employed. The theoretical method has been utilized to predict the beryllium chemical shifts of structurally characterized complexes for which no measured (9)Be NMR spectrum exists, and to investigate a literature complex with an unusual (9)Be NMR chemical shift. A new standard for beryllium NMR in nonaqueous solvents has been suggested. PMID- 15521786 TI - Opening mechanism of G.T/U pairs in DNA and RNA duplexes: a combined study of imino proton exchange and molecular dynamics simulation. AB - The opening pathway of wobble pairs dG.T and rG.U has been investigated in four DNA and two RNA duplexes. Using NMR spectroscopy, we measured the imino proton exchange of both G(H1) and T/U(H3), catalyzed by ammonia, tris, and OH(-), and we calculated the free energy surface related to G.T/U opening by molecular dynamics simulations. Taken together the experimental and theoretical results, we suggest that wobble pairs open through a coupled rotation of the bases toward the major groove where exchange of both imino protons takes place with the surrounding water. PMID- 15521787 TI - Key role of electrostatic interactions in bacteriorhodopsin proton transfer. AB - The first proton transport step following photon absorption in bacteriorhodopsin is from the 13-cis retinal Schiff base to Asp85. Configurational and energetic determinants of this step are investigated here by performing quantum mechanical/molecular mechanical minimum-energy reaction-path calculations. The results suggest that retinal can pump protons when in the 13-cis, 15-anti conformation but not when 13-cis, 15-syn. Decomposition of the proton transfer energy profiles for various possible pathways reveals a conflict between the effect of the intrinsic proton affinities of the Schiff base and Asp85, which favors the neutral, product state (i.e., with Asp85 protonated), with the mainly electrostatic interaction between the protein environment with the reacting partners, which favors the ion pair reactant state (i.e., with retinal protonated). The rate-limiting proton-transfer barrier depends both on the relative orientations of the proton donor and acceptor groups and on the pathway followed by the proton; depending on these factors, the barrier may arise from breaking and forming of hydrogen bonds involving the Schiff base, Asp85, Asp212, and water w402, and from nonbonded interactions involving protein groups that respond to the charge rearrangements in the Schiff base region. PMID- 15521788 TI - Combination therapy in Alzheimer's disease: a review of current evidence. AB - Treating dementia has become a major challenge in clinical practice. Presently, acetylcholinesterase inhibitors are the first-line drugs in the treatment of Alzheimer's disease (AD). These options are now complemented by memantine, which is approved for the treatment of moderate-to-severe AD. Altogether, a minimum of six agent classes already exist, all of which are approved for clinical use and are either already being tested or ready for phase III clinical trials for the treatment of AD. These include cholinesterase inhibitors, blockers of the NMDA receptor, antioxidants or blockers of oxidative deamination (including Gingko biloba), anti-inflammatory agents, neurotrophic factors (including hormone replacement therapy and drugs acting on insulin signal transduction) and antiamyloid agents (including cholesterol-lowering therapy). These approaches hold promise for disease modification and have a potential to be used as combination therapy for cognitive enhancement. Presently, only nine clinical studies have been published that have investigated the effects of a combination regimen on cognitive performance or AD. Among those, one study was conducted in elderly cognitively intact persons; the others involved patients with AD. Only five of the treatment studies followed a randomised, controlled design. Not all studies favoured the superior efficacy of combination therapy over monotherapy. Some studies, however, showed some evidence for synergistic combination effects of symptomatic therapy, including delay or prevention of disease progression in AD patients. In addition, six studies investigated the effects of AChE inhibitor in combination with antipsychotic or antidepressant therapy on behavioural aspects of AD symptomatology. In four of those studies there were indications that combination therapy had greater efficacy over monotherapy. The treatment of AD patients requires optimised options for all stages of illness based on the available drugs. There is a great need for further well designed studies on combination therapy in AD. PMID- 15521789 TI - Starting clozapine in the community: a UK perspective. AB - Clozapine is the most effective antipsychotic available for the treatment of schizophrenia that has proved resistant to other medications and the only antipsychotic licensed for this indication. Although the drug is increasingly being used more widely in patients with schizophrenia and with other psychiatric and neurological disorders, it is still underused. The main reasons for this are that it can cause adverse effects such as weight gain and sedation, and the need for regular blood test monitoring because of the risk of agranulocytosis. While these hurdles are unavoidable, another problem in the UK has been the historical practice of admitting patients to hospital to initiate treatment with clozapine. However, protocols have now been developed for both home and day-hospital initiation. The experience of one assertive community treatment team of starting clozapine in patients' own homes has been positive, with no major adverse events reported. This approach is, however, extremely demanding of staff resources and for many services the use of day-hospitals to initiate treatment with clozapine is more appropriate. Research into staff and patients' views about community initiation of clozapine, and its economic costs, would be welcome. PMID- 15521790 TI - Activities of daily living in patients with dementia: clinical relevance, methods of assessment and effects of treatment. AB - Disability, characterised by the loss of ability to perform activities of daily living (ADL), is a defining feature of dementia that results in growing caregiver burden and the eventual need for alternative care or nursing home placement. Functional decline in patients with dementia can also result from causes other than dementia, such as comorbid medical and psychiatric illnesses and sensory impairment. ADL consists of instrumental ADL (IADL) [complex higher order skills, such as managing finances] and basic ADL (BADL) [self-maintenance skills, such as bathing]. Assessment of IADL and BADL is recommended to establish a diagnosis of dementia. Functional assessment also helps the healthcare provider to offer appropriate counselling regarding safety concerns and need for custodial care. Functional capacity measures have been used increasingly in pharmacological trials of patients with Alzheimer's disease (AD) and related dementias, although at the present time these measures are generally not primary outcome measures. Functional impairment is not a uniform construct; rather, it is multifaceted and can be measured with various clinical instruments. Many scales have been validated for use in patients with AD for characterising functional impairment and evaluating the efficacy of treatment. Research to date indicates that cholinesterase inhibitors have the potential for modest but meaningful beneficial effects on ADL in patients with mild-to-moderate AD. Memantine also has promising beneficial effects on functional abilities in persons with moderate-to-severe AD. Assessment of ADL as a primary efficacy measure using a validated scale that is non-gender biased and cross-nationally relevant is recommended in new treatment trials of patients with AD and related dementias. PMID- 15521791 TI - Impact of atypical antipsychotics on quality of life in patients with schizophrenia. AB - Schizophrenia is a long-term disabling illness that affects approximately 1% of the population. Its course is generally chronic with acute psychotic exacerbations that may require frequent hospitalisations. The clinical picture includes a range of symptoms such as delusions, hallucinations, agitation, suspiciousness, hostility, conceptual disorganisation, blunted affect, emotional and social withdrawal, lack of spontaneity, poverty of speech and a wide range of neurocognitive deficits. Over the past 50 years, antipsychotic medications have emerged as the cornerstone of management in concert with other important interventions, such as psychosocial and economic support and rehabilitation efforts. However, the unrivalled role of conventional antipsychotic medications has been continuously challenged by the wide range of adverse effects of these medications and their lack of usefulness in the treatment of neurocognitive deficits as well as deficit and negative symptoms. In addition, the lack of subjective tolerability of these agents and their negative impact on quality of life have complicated management for a large number of patients. Over the last 15 years, several new atypical antipsychotic medications have been introduced, including amisulpride, remoxipride, risperidone, sertindole, olanzapine, zotepine, quetiapine, ziprasidone and aripiprazole. In general, the new antipsychotics have shown themselves to be at least comparable in efficacy to conventional antipsychotics but with superior subjective tolerability and a more favourable adverse effect profile. The majority of quality of life studies involving new antipsychotic agents have evaluated the benefits of risperidone, olanzapine and clozapine; only a few studies have examined the effects of other new antipsychotics. While most of these studies have methodological and design limitations, the weight of evidence from them nevertheless points to a trend towards a more positive impact on quality of life with atypical agents. A number of recommendations can be made. First, more independent well designed and controlled studies are urgently needed to evaluate the effects of antipsychotic therapy on quality of life in patients with schizophrenia. New comparative studies should explore not only the differences between new and old antipsychotics but also identify any potential differences between individual new agents. The role of cost-effectiveness studies such as cost utility approaches in schizophrenia needs to be revisited, notwithstanding the fact that these types of studies have been reported to be feasible in schizophrenia. Finally, quality-of life-based pharmacoeconomic studies of antipsychotic agents should not concentrate solely on cost reduction or containment, as it is likely that in order to maximise the benefits of new antipsychotic medications, greater expenditure on rehabilitation programmes and other support services will be necessary in the short-term at least. PMID- 15521793 TI - A comparison of the direct costs and cost effectiveness of serotonin reuptake inhibitors and associated adverse drug reactions. AB - BACKGROUND: The economic burden of depression is known to be high and was estimated to be USD 83.1 billion in 2000. Serotonin reuptake inhibitors (SRIs), including both selective serotonin reuptake inhibitors (SSRIs) and serotonin norepinephrine reuptake inhibitors (SNRIs), have a superior adverse effect and safety profile relative to traditional agents (e.g. TCAs), and as a result have demonstrated superior cost effectiveness. Although efficacy across the SRIs is similar, the incidence of adverse drug reactions (ADRs) within SRIs remains significant and varies by agent. Patients who experience ADRs from SRIs may seek medical care, require additional treatment, and even discontinue treatment altogether, leading to increased utilisation and cost of therapy. OBJECTIVE: This study estimates the direct cost and cost effectiveness, taking into account the impact of treatment-related ADRs, of eight currently marketed SRIs (citalopram, escitalopram, generic fluoxetine, paroxetine, paroxetine controlled release [CR], sertraline, venlafaxine and venlafaxine extended release [XR]) used as initial treatment for depression. METHODS: A decision analytic model with a 6-month treatment goal was used to estimate the direct cost and cost effectiveness of treatment from the managed care/payer perspective. Estimates of SRI-related ADRs, associated treatments and costs were derived from the US FDA-approved prescribing information and published literature. Efficacy was assumed to be similar across all SRIs. Effectiveness was measured using quality-adjusted life years (QALY) based on EuroQol EQ-5D scores derived from the 2000 Medical Expenditure Panel Survey (MEPS). Censored least absolute deviations (CLAD) regression analysis was used to derive age-adjusted estimates of utility for all health states. Univariate and Bayesian second-order multivariate probabilistic sensitivity analyses were conducted to examine the impact of uncertainty in the parameter estimates. RESULTS: The expected direct cost and cost effectiveness of treatment from least to most expensive were: escitalopram (USD 3891; 0.341), citalopram (USD 3938; 0.340), generic fluoxetine (USD 4034; 0.335), venlafaxine XR (USD 4226; 0.336), sertraline (USD 4250; 0.335), generic paroxetine (USD 4385; 0.332), paroxetine CR (USD 4440; 0.332) and venlafaxine (USD 4613; 0.326). Monte Carlo simulation results suggested that escitalopram was the most likely (77%) to be cost effective for a willingness to pay < or = USD 50,000 per QALY, followed by citalopram (22%), generic fluoxetine (0.3%) and all other SRIs (0%). Sensitivity analyses indicated that the results of the study were robust to the assumptions underpinning the model. CONCLUSIONS: SRI-related ADRs have a significant impact on the direct cost and cost effectiveness of treatment. Escitalopram, with the lowest ADR rate of the SRIs, had the lowest expected treatment cost and greatest effectiveness when compared with citalopram, generic fluoxetine, generic paroxetine, paroxetine CR, sertraline, venlafaxine and venlafaxine XR. PMID- 15521792 TI - Cannabis-associated psychosis: current status of research. AB - Cannabis has been used for recreational, medicinal and religious purposes in different cultures since ancient times. There have been various reports of adverse effects due to or associated with cannabis consumption, including psychotic episodes. Historically, our understanding of these clinical observations has been significantly hindered by a lack of knowledge regarding their underlying neurobiological and pharmacological processes. However, the discovery of the endogenous cannabinoid system has allowed a greater understanding of these adverse effects to develop. From a clinical perspective, toxic or transient psychotic reactions to the administration of herbal cannabis preparations or specific cannabinoid compounds have to be differentiated from longer-lasting, persistent schizophrenia-like disorders associated with the use of cannabis/cannabinoids. The latter are most likely to be associated with a predisposition or vulnerability to schizophrenia. Interestingly, the recently suggested role of the endogenous cannabinoid system in schizophrenia not related to previous cannabinoid consumption introduces an additional perspective on the mechanism underlying cannabis-associated schizophrenia-like disorders, as well as on the effects of cannabis consumption in schizophrenia. At present, acute psychopharmacological treatment options for cannabis-associated transient and persistent schizophrenia-like psychotic episodes are similar and are based on the use of benzodiazepines and antipsychotics. However, new pharmacological strategies using the endogenous cannabinoid system as a primary target are under development. Long-term psychotherapeutic treatment options involve case management strategies and are mainly based on specialised psychotherapeutic programmes to encourage cannabis users to stop their use of the drug. PMID- 15521795 TI - Response reversals in recognition memory. AB - Using a dynamic sequential sampling model and a recently proposed model for confidence judgments in recognition memory (T. Van Zandt, 2000b), the authors examine the tendency for rememberers to reverse their responses after a primary decision. In 4 experiments, speeded "old"-"new" decisions were made under bias followed by a 2nd response', either a confidence judgment or another simple choice. The data from these experiments showed that participants know when they have made a mistake and that they respond to this knowledge by reversing their responses. Response reversals are thus shown to be important for constructing models of the response-selection process in recognition memory. PMID- 15521794 TI - Amisulpride: a review of its use in the management of schizophrenia. AB - Amisulpride (Solian), a substituted benzamide derivative, is a second-generation antipsychotic that preferentially binds to dopamine D2/D3 receptors in limbic rather than striatal structures. High dosages preferentially antagonise postsynaptic D2/D3 receptors, resulting in reduced dopamine transmission, and low dosages preferentially block presynaptic D2/D3 receptors, resulting in enhanced dopamine transmission. Amisulpride (200-1200 mg/day) was at least as effective as haloperidol and as effective as risperidone or olanzapine, in studies of up to 1 year in patients with schizophrenia manifesting predominantly positive symptoms. Amisulpride (50-300 mg/day) was significantly more effective than placebo in studies of up to 6 months in patients manifesting predominantly negative symptoms. Quality of life was also improved significantly more in patients receiving amisulpride than in those receiving haloperidol in 4- and 12-month studies in patients with predominantly mixed symptoms. Amisulpride was generally well tolerated in clinical trials. In patients with predominantly positive symptoms, amisulpride appeared to be better tolerated than haloperidol and was tolerated as well as risperidone and olanzapine. The incidence of extrapyramidal adverse effects with amisulpride was lower than with haloperidol but was generally similar to risperidone or olanzapine. Weight gain with amisulpride was less than that with risperidone or olanzapine and, unlike these agents, amisulpride does not seem to be associated with diabetogenic effects. Plasma prolactin levels are increased during amisulpride therapy and amenorrhoea occurs in about 4% of women. The incidence of adverse events with low dosages of amisulpride (< or = 300 mg/day) in patients with predominantly negative symptoms was similar to that observed with placebo. In conclusion, oral amisulpride (200 1200 mg/day) is at least as effective as haloperidol, and as effective as risperidone or olanzapine, in the treatment of patients with schizophrenia manifesting predominantly positive symptoms. In the treatment of patients manifesting predominantly negative symptoms, low dosages of amisulpride (50-300 mg/day) are significantly more effective than placebo. Amisulpride appears to be better tolerated than haloperidol, causing a lower incidence of extrapyramidal adverse effects and an improved quality of life. Compared with risperidone or olanzapine, amisulpride is more likely to cause hyperprolactinaemia, but has a lower propensity to cause weight gain and does not seem to be associated with diabetogenic effects. Thus, amisulpride is an effective and well tolerated option for the first-line treatment of patients with acute schizophrenia as well as for those requiring long-term maintenance therapy. PMID- 15521796 TI - Subjective probability intervals: how to reduce overconfidence by interval evaluation. AB - Format dependence implies that assessment of the same subjective probability distribution produces different conclusions about over- or underconfidence depending on the assessment format. In 2 experiments, the authors demonstrate that the overconfidence bias that occurs when participants produce intervals for an uncertain quantity is almost abolished when they evaluate the probability that the same intervals include the quantity. The authors successfully apply a method for adaptive adjustment of probability intervals as a debiasing tool and discuss a tentative explanation in terms of a naive sampling model. According to this view, people report their experiences accurately, but they are naive in that they treat both sample proportion and sample dispersion as unbiased estimators, yielding small bias in probability evaluation but strong bias in interval production. PMID- 15521797 TI - Six regularities of source recognition. AB - In recent work, researchers have shown that source-recognition memory can be incorporated in an extended signal detection model that covers both it and item recognition memory (A. Hilford, M. Glanzer, K. Kim, & L. T. DeCarlo, 2002). In 5 experiments, using learning variables that have an established effect on item recognition, the authors tested further implications of that extended model. The results establish 6 source-recognition regularities that support the model. An important regularity in that set is the linkage of source and item recognition: Any learning variable that increases item recognition increases source recognition. PMID- 15521798 TI - The effect of the length of to-be-remembered lists and intervening lists on free recall: a reexamination using overt rehearsal. AB - In 3 experiments, the authors investigated the effects of to-be-remembered (TBR) and intervening list length on free recall to determine whether selective rehearsal could explain the previous finding that recall was affected only by TBR list length. In Experiments 1 (covert rehearsal) and 2 (overt rehearsal), participants saw 5- and 20-word lists and had to recall the list prior to that last presented list. In Experiment 3, either 1 or 2 lists were presented, and recall of TBR list was postcued. Recall proportion decreased with increased TBR list length. Moreover, the authors found extended recency effects when recall was replotted by when words were last rehearsed (Experiments 2 and 3) and an effect of intervening list length when rehearsal was reduced (Experiment 3). PMID- 15521799 TI - The metrics of spatial distance traversed during mental imagery. AB - The authors conducted 2 experiments to study the metrics of spatial distance in a mental imagery task. In both experiments, participants first memorized the layout of a building containing 10 rooms with 24 objects. Participants then received mental imagery instructions and imagined how they walked through the building from one room to another. The authors manipulated Euclidean distance involved in these imaginary motions: Spatial distance measured in centimeters on the layout was either short or long. Independently, they varied categorical distance: The motions led through one room or two rooms. The time needed to imagine motions and response times to test probes indicated that both Euclidean distance and categorical distance affected mental imagery. The authors discuss the new finding of categorical distance effects in mental imagery and relate the results to earlier failures to find Euclidean distance effects in formally equivalent studies of narrative comprehension. PMID- 15521800 TI - Immediate and sustained effects of planning in a problem-solving task. AB - In 4 experiments, instructions to plan a task (water jugs) that normally produces little planning altered how participants solved the problems and resulted in enhanced learning and memory. Experiment 1 identified planning strategies that allowed participants to plan full solutions to water jugs problems. Experiment 2 showed that experience with planning led to better solutions even after planning was no longer required, whereas control participants showed little improvement. Experiments 3 and 4 showed that although the most recent planned solution could be recalled following a long filled retention interval, retroactive interference (RI) between successive problems resulted in much lower recall of earlier solutions. RI during plan generation could also explain participants' choice of depth-first planning strategies. PMID- 15521801 TI - Intentional control of event counting. AB - Event counting depends on simple, well-learned knowledge but is effortful and error-prone. In 6 experiments, the authors examined event-counting performance, testing a model that suggests that counting is controlled by minimal goal representations coordinated with perceptual events by temporal synchrony. In Experiment 1, they examined self-paced counting with or without delays that disrupted participants' preferred pacing. In subsequent experiments, participants counted computer-paced events occurring at rhythmic or varied intervals, reporting or verifying totals. Several results support the model: Participants counted rhythmic events more accurately, made undetected undercount errors when counting rhythmic events, and made false alarms to undercount or overcount probes presented at different times. These results suggest that intentions that guide fluent counting specify parameters deictically rather than semantically and that error monitoring is implicit. PMID- 15521802 TI - Semantic ambiguity and the process of generating meaning from print. AB - An ambiguity disadvantage (slower responses for ambiguous words, e.g., bank, than for unambiguous words) has been reported in semantic tasks (L. R. Gottlob, S. D. Goldinger, G. O. Stone, & G. C. Van Orden, 1999; Y. Hino, S. J. Lupker, & P. M. Pexman, 2002; C. D. Piercey & S. Joordens, 2000) and has been attributed to the meaning activation process. The authors tested an alternative explanation; The ambiguity disadvantage arises from the decision-making process in semantic tasks. The authors examined effects of ambiguity on unrelated trials in a relatedness decision task, because these trials are free from response competition created by ambiguous words on related trials. Results showed no ambiguity effect on unrelated trials (Experiments 2, 3c, and 5c) and an ambiguity disadvantage on related trials (Experiments 3a, 3b, 5a, and 5b). PMID- 15521803 TI - Morphological family size in a morphologically rich language: the case of Finnish compared with Dutch and Hebrew. AB - Finnish has a very productive morphology in which a stem can give rise to several thousand words. This study presents a visual lexical decision experiment addressing the processing consequences of the huge productivity of Finnish morphology. The authors observed that in Finnish words with larger morphological families elicited shorter response latencies. However, in contrast to Dutch and Hebrew, it is not the complete morphological family of a complex Finnish word that codetermines response latencies but only the subset of words directly derived from the complex word itself. Comparisons with parallel experiments using translation equivalents in Dutch and Hebrew showed substantial cross-language predictivity of family size between Finnish and Dutch but not between Finnish and Hebrew, reflecting the different ways in which the Hebrew and Finnish morphological systems contribute to the semantic organization of concepts in the mental lexicon. PMID- 15521804 TI - When words collide: facilitation and interference in the report of repeated words from rapidly presented lists. AB - Inhibited encoding is the basis of some accounts of repetition blindness- impaired report of the second occurrence of a repeated word in a rapidly presented word sequence. The author presents evidence for the claim that repetition effects arise from constructive processes of perception and memory that occur to some extent after the word sequence has been presented. Unpredictable postlist cues prompted subjects to report either the entire list or just the final word in the list. Repetition impaired the report of the second occurrence of a repeated word under full report but facilitated the report of such items when only the final word had to be reported. The author modulated this dissociation by presenting repeated words in sentences rather than unrelated word lists. The sensitivity of the effects of repetition to postlist cues supports a construction rather than an encoding inhibition account of repetition blindness. PMID- 15521805 TI - The effect of plausibility on eye movements in reading. AB - Readers' eye movements were monitored as they read sentences describing events in which an individual performed an action with an implement. The noun phrase arguments of the verbs in the sentences were such that when thematic assignment occurred at the critical target word, the sentence was plausible (likely theme), implausible (unlikely theme), or anomalous (an inappropriate theme). Whereas the target word in the anomalous condition provided evidence of immediate disruption, the effect of the target word in the implausible condition was considerably delayed. The results thus indicate that when a word is anomalous, it has an immediate effect on eye movements, but that the effect of implausibility is not as immediate. PMID- 15521806 TI - Working memory capacity and the antisaccade task: individual differences in voluntary saccade control. AB - Performance on antisaccade trials requires the inhibition of a prepotent response (i.e., don't look at the flashing cue) and the generation and execution of a correct saccade in the opposite direction. The authors attempted to further specify the role of working memory (WM) span differences in the antisaccade task. They tested high- and low-span individuals on variants of prosaccade and antisaccade trials in which an eye movement is the sole requirement. In 3 experiments, they demonstrated the importance of WM span differences in both suppression of a reflexive saccade and generation of a volitional eye movement. The results support the contention that individual differences in WM span are not exclusively due to differences in inhibition but also reflect differences in directing the focus of attention. PMID- 15521807 TI - A working memory workout: how to expand the focus of serial attention from one to four items in 10 hours or less. AB - Five individuals participated in an extensive practice study (10 1-hr sessions, 11,000 trials total) on a self-paced identity-judgment (1)n-back task (n ranging from 1 to 5). Within Session 1, response time increased abruptly by about 300 ms in passing from n = 1 to n > 1, suggesting that the focus of attention can accommodate only a single item (H. Caravan, 1998; B. McElree, 2001). Within Session 10, response time was dramatically reduced and increased linearly with n for n < or = 4, with a slope of about 30 ms. The data suggest that working memory consists of a focus of attention governed by a limited-capacity search, expandable through practice, and a content-addressable region outside the focus of attention. PMID- 15521808 TI - Folding and stability of chimeric immunofusion VL-barstar. AB - A chimeric protein VL-barstar that comprises the VL domain of anti-human ferritin monoclonal antibody F11 and barstar, the naturally occurring inhibitor of bacterial RNase barnase, has been constructed for study of structure-function characteristics of chimeric immunoglobulin fused proteins. Such chimeric constructs may be potentially employed for development of bivalent/bispecific antibodies on the basis of the high affinity interaction between barstar and barnase (the association constant is about 10(14) M(-1)). We have developed a protocol for VL-barstar expression in E. coli and purification and refolding from inclusion bodies that yields a homogeneous and soluble form of this protein. Differential scanning calorimetry in combination with fluorescence and CD spectroscopy revealed that the VL-barstar formed well-resolved ordered secondary and compact tertiary structures. However, partial loss of tertiary interactions resulted in low stability of the recombinant protein and the lack of functional activity of the two constituent modules. These conformational features suggest that the protein might be referred to the class of native molten globules, which comprises partially unfolded conformations stabilized under physiological conditions. Since individually expressed VL domain and barstar retain completely folded conformation and stable spatial structure, the incomplete folding of the chimeric protein may be attributed to interaction between heterologous domains, which appears at the folding stage preceding formation of a system of tertiary interactions in both structural modules. The results provide evidence for non native interactions between heterologous modules that may occur in chimeric proteins composed of taxonomically distinct fusion partners. PMID- 15521809 TI - Catalase and superoxide dismutase: distribution, properties, and physiological role in cells of strict anaerobes. AB - This review considers the distribution of the main enzymes of antioxidative defense, superoxide dismutase (SOD) and catalase, in various groups of strictly anaerobic microorganisms: bacteria of the genus Clostridium, Bacteroides, sulfate reducing and acetogenic bacteria, methanogenic archaea, etc. Molecular and biochemical properties of purified Fe-containing SODs, cambialistic SODs, and heme catalases are presented. The physiological role and origin of the enzymes of antioxidative defense in strict anaerobes are discussed. Physiological responses (induction of SOD and catalase) to factors provoking oxidative stress in the cells of strict anaerobes able to maintain viability under aerobic conditions are also considered. PMID- 15521810 TI - Optical characteristics of thiamine in model systems and in holoenzyme. AB - The optical properties of thiamine diphosphate-dependent enzymes change significantly on their interaction with cofactors (thiamine, bivalent metal ions) and substrates. These changes are connected with structural alterations of the active site and the mechanism of its functioning, and in some cases they reflect changes in the optical properties of the coenzyme itself within the protein. The use of optical characteristics, especially together with model systems, appeared to be a rather promising approach for investigation of the active site of thiamine diphosphate-dependent enzymes and the mechanism of its functioning. So, it seemed to be useful to summarize the literature data concerning the optical characteristics of thiamine (thiamine diphosphate) in model systems and the efficiency of their application for study of thiamine diphosphate-dependent enzymes. PMID- 15521811 TI - Protein folding, misfolding, and aggregation. Formation of inclusion bodies and aggresomes. AB - In this review the mechanisms of protein folding, misfolding, and aggregation as well as the mechanisms of cell defense against toxic protein aggregates are considered. Misfolded and aggregated proteins in cells are exposed to chaperone mediated refolding and are degraded by proteasomes if refolding is impossible. Proteolysis-stable protein aggregates accumulate, forming inclusion bodies. In eucaryotic cells, protein aggregates form structures in the pericentrosomal area that have been termed "aggresomes". Formation of aggresomes in cells is a general cellular response to the presence of misfolded proteins when the degrading capacity of the cells is exceeded. The role of aggresomes in disturbance of the proteasomal system operation and in cellular death, particularly in the so-called "protein conformational diseases", is discussed. PMID- 15521812 TI - Oxidation of phenolic compounds by peroxidase in the presence of soluble polymers. AB - The kinetics of Coprinus cinereus peroxidase-catalyzed 1-naphthol, 2-naphthol, and 4-hydroxybiphenyl oxidation was investigated. The initial rates of the naphthols' and 4-hydroxybiphenyl oxidations were linearly dependent on enzyme concentration. The rates depended on substrate concentration and saturated at concentrations above 100 microM of hydrogen peroxide, 25-50 microM of naphthols, and 10 microM of 4-hydroxybiphenyl. At the peroxide concentration 100 microM calculated K(m) and the maximal rate (V(max)) were 74.7 microM and 0.53 microM/sec or 175 microM and 2.0 microM/sec for 1- or 2-naphthol, respectively, and 29.68 microM and 0.42 microM/sec for 4-hydroxybiphenyl. Kinetic measurements of exhaustive naphthol and 4-hydroxybiphenyl oxidation showed that peroxidase is inactivated during the oxidation of the substrates. Different factors and additives, water soluble polymers and albumins (PEG, PEI, PL, BSA, HSA), influenced the initial naphthols and 4-hydroxybiphenyl oxidation rates, peroxidase inactivation rates, and the degree of the substrate conversion. Addition of albumin increased turnover number of naphthols oxidation 1.5-4 times. Light scattering increase was observed when peroxidase-catalyzed oxidation reaction was investigated and suggested that insoluble particles were formed during the process. The addition of polymers, change of concentration and ionic strength of the solution as well as the number of other factors influenced the observed light scattering. The number of particles formed during peroxidase catalyzed naphthols' and 4-hydroxybiphenyl oxidation and their distribution according to size in the interval 2.5-300 microm were detected by particle counting in solutions. PMID- 15521813 TI - Biochemical characterization and distribution of glutathione S-transferases in leaping mullet (Liza saliens). AB - In this study, feral leaping mullet (Liza saliens) liver cytosolic glutathione S transferases (GSTs) were investigated and characterized using 1-chloro-2,4 dinitrobenzene (CDNB) and ethacrynic acid (EA) as substrates. The average GST activities towards CDNB and EA were found to be 1365 +/- 41 and 140 +/- 20 nmol/min per mg protein, respectively. The effects of cytosolic protein amount and temperature ranging from 4 to 70 degrees C on enzyme activities were examined. While both activities towards CDNB and EA showed similar dependence on protein amount, temperature optima were found as 37 and 42 degrees C, respectively. In addition, the effects of pH on GST-CDNB and -EA activities were studied and different pH activity profiles were observed. For both substrates, GST activities were found to obey Michaelis-Menten kinetics with apparent V(max) and K(m) values of 1661 nmol/min per mg protein and 0.24 mM and 157 nmol/min per mg protein and 0.056 mM for CDNB and EA, respectively. Distribution of GST in Liza saliens tissues was investigated and compared with other fish species. Very high GST activities were measured in tissues from Liza saliens such as liver, kidney, testis, proximal intestine, and gills. Moreover, our results suggested that GST activities from Liza saliens would be a valuable biomarker for aquatic pollution. PMID- 15521814 TI - Neuroprotection by alpha-lipoic acid in streptozotocin-induced diabetes. AB - Glial cells provide structural and metabolic support for neurons, and these cells become reactive to any insult to the central nervous system. The streptozotocin (STZ) rat model was used to study glial reactivity and the prevention of gliosis by alpha-lipoic acid (alpha-LA) administration. The expression of glial fibrillary acidic protein (GFAP), S100B protein, and neuron specific enolase (NSE) was determined as well as lipid peroxidation (LPO) and glutathione (GSH) levels in some brain tissues. Western blot analyses showed GFAP, S100B, and NSE levels significantly increased under STZ-induced diabetes in brain, and LPO level increased as well. Administration of alpha-LA reduced the expression both of glial and neuronal markers. In addition, alpha-LA significantly prevented the increase in LPO levels found in diabetic rats. GSH levels were increased by the administration of alpha-LA. This study suggests that alpha-LA prevents neural injury by inhibiting oxidative stress and suppressing reactive gliosis. PMID- 15521815 TI - Lactoferrin is the major deoxyribonuclease of human milk. AB - Lactoferrin is the major iron-transferring protein of human barrier fluids such as blood and milk. It is a polyfunctional protein capable of binding DNA exposed on the surface of various cells. Electrophoretically homogenous lactoferrin was prepared by sequential chromatography of human milk proteins on DEAE-cellulose, heparin-Sepharose, and Sepharose containing immobilized anti-lactoferrin antibodies. By subsequent chromatography on Blue Sepharose the resulting lactoferrin was fractionated into several subfractions with different affinity for the sorbent, and this was associated with separation of additional lactoferrin peaks with DNase activity from the main peak. By various techniques, in particular, by in situ testing the DNase activity of lactoferrin in a DNA containing gel after SDS-electrophoresis, hydrolysis of DNA was for the first time shown to be an intrinsic property of lactoferrin. The substrate specificity of lactoferrin in hydrolysis of DNA was different from specificities of known human DNases. Hydrolysis of DNA was activated by bivalent metal ions and also by ATP and NAD. Unlike the main fraction of lactoferrin with the highest affinity for Blue Sepharose, all protein subfractions with DNase activity were cytotoxic and suppressed growth of human and mouse tumor cell lines. PMID- 15521816 TI - Effects of diffusion and topological factors on the efficiency of energy coupling in chloroplasts with heterogeneous partitioning of protein complexes in thylakoids of grana and stroma. A mathematical model. AB - In this work, we studied theoretically the effects of diffusion restrictions and topological factors that could influence the efficiency of energy coupling in the heterogeneous lamellar system of higher plant chloroplasts. Our computations are based on a mathematical model for electron and proton transport in chloroplasts coupled to ATP synthesis in chloroplasts that takes into account the nonuniform distribution of electron transport and ATP synthase complexes in the thylakoids of grana and stroma. Numerical experiments allowed the lateral profiles of pH in the thylakoid lumen and in the narrow gap between grana thylakoids to be simulated under different metabolic conditions (in the state of photosynthetic control and under conditions of photophosphorylation). This model also provided an opportunity to simulate the effects of steric constraints (the extent of appression of thylakoids in grana) on the rates of non-cyclic electron transport and ATP synthesis. This model demonstrated that there might be two mechanisms of regulation of electron and proton transport in chloroplasts: 1) slowing down of non-cyclic electron transport due to a decrease in the intra-thylakoid pH, and 2) retardation of plastoquinone reduction due to slow diffusion of protons inside the narrow gap between the thylakoids of grana. Numerical experiments for model systems that differ with respect to the arrangement of thylakoids in grana allowed the effects of osmolarity on the photophosphorylation rate in chloroplasts to be explained. PMID- 15521817 TI - Calcium-dependent nonspecific permeability of the inner mitochondrial membrane is not induced in mitochondria of the yeast Endomyces magnusii. AB - Mitochondria of the yeast Endomyces magnusii were examined for the presence of a Ca2+- and phosphate-induced permeability of the inner mitochondrial membrane (pore). For this purpose, coupled mitochondria were incubated under conditions known to induce the permeability transition pore in animal mitochondria, i.e., in the presence of high concentrations of Ca2+ and P(i), prooxidants (t butylhydroperoxide), oxaloacetate, atractyloside (an inhibitor of ADP/ATP translocator), SH-reagents, by depletion of adenine nucleotide pools, and deenergization of the mitochondria. Large amplitude swelling, collapse of the membrane potential, and efflux of the accumulated Ca2+ were used as parameters for demonstrating pore induction. E. magnusii mitochondria were highly resistant to the above-mentioned substances. Deenergization of mitochondria or depletion of adenine nucleotide pools have no effect on low-amplitude swelling or the other parameters. Cyclosporin A, a specific inhibitor of the nonspecific permeability transition in animal mitochondria, did not affect the parameters measured. It is thus evident that E. magnusii mitochondria lack a functional Ca2+-dependent pore, or possess a pore differently regulated as compared to that of mammalian mitochondria. PMID- 15521818 TI - New structures of the O-specific polysaccharides of proteus. 4. Polysaccharides containing unusual acidic N-acyl derivatives of 4-amino-4,6-dideoxy-D-glucose. AB - The structures of the O-polysaccharides of the lipopolysaccharides of Proteus mirabilis O7 and O49 were determined by chemical methods, mass spectrometry, including MS/MS, and NMR spectroscopy, including experiments run in an H2O/D2O mixture to reveal correlations for NH protons. The O-polysaccharides were found to contain N-carboxyacetyl (malonyl) and N-(3-carboxypropanoyl) (succinyl) derivatives of 4-amino-4,6-dideoxyglucose (4-amino-4-deoxyquinovose, Qui4N), respectively. The behavior of Qui4N derivatives with the dicarboxylic acids under conditions of acid hydrolysis and methanolysis was studied using GLC-MS. PMID- 15521819 TI - Positioning of a nucleosome on mouse satellite DNA inserted into a yeast plasmid is determined by its DNA sequence and an adjacent nucleosome. AB - It has earlier been shown that multiple positioning of nucleosomes on mouse satellite DNA is determined by its nucleotide sequence. To clarify whether other factors, such as boundary ones, can affect the positionings, we modified the environment of satellite DNA monomer by inserting it into a yeast plasmid between inducible GalCyc promoter and a structural region of the yeast FLP gene. We have revealed that the positions of nucleosomes on satellite DNA are identical to those detected upon reconstruction in vitro. The positioning signal (GAAAAA sequence) of satellite DNA governs nucleosome location at the adjacent nucleotide sequence as well. Upon promoter induction the nucleosome, translationally positioned on the GalCyc promoter, transfers to the satellite DNA and its location follows the positioning signal of the latter. Thus, the alternatives of positioning of a nucleosome on satellite DNA are controlled by its nucleotide sequence, though the choice of one of them is determined by the adjacent nucleosome. PMID- 15521821 TI - Filopodia formation and Disabled degradation downstream of Reelin. AB - During Drosophila embryogenesis, Abl (Abelson tyrosine kinase) is localized in the axons of the CNS (central nervous system). Mutations in Abl have a subtle effect on the morphology of the embryonic CNS, and the mutant animals survive to the pupal and adult stages. However, genetic screens have identified several genes that, when mutated along with the Abl gene, modified the phenotypes. Two prominent genes that arose from these screens were enabled (Ena) and disabled (Dab). It has been known for some time that Enabled and its mammalian homologues are involved in the regulation of actin dynamics, and promote actin polymerization at the leading edge of motile cells. It was a defect in actin polymerization in migrating neurons in particular that resulted in the identification of Enabled as an important regulator of neuronal migration. Defects in Disabled, in both Drosophila and mammals, also gave rise to neuronal defects which, in mice, were indistinguishable from phenotypes observed in the reeler mouse. These observations suggested that mDab1 (mammalian Disabled homologue 1) acted in a pathway downstream of Reelin, the product of the reelin gene found to be defective in reeler mice. Now, in this issue of the Biochemical Journal, Takenawa and colleagues have demonstrated that Disabled also acts in a pathway to regulate actin dynamics through the direct activation of N-WASP (neuronal Wiskott-Aldrich syndrome protein). Furthermore, they were also able to link several lines of investigation from other groups to show that the ability of mDab1 to regulate actin dynamics during cell motility was under the negative control of tyrosine phosphorylation, leading to ubiquitin-mediated degradation of mDab1. PMID- 15521820 TI - Essential tension and constructive destruction: the spindle checkpoint and its regulatory links with mitotic exit. AB - Replicated genetic material must be partitioned equally between daughter cells during cell division. The precision with which this is accomplished depends critically on the proper functioning of the mitotic spindle. The assembly, orientation and attachment of the spindle to the kinetochores are therefore constantly monitored by a surveillance mechanism termed the SCP (spindle checkpoint). In the event of malfunction, the SCP not only prevents chromosome segregation, but also inhibits subsequent mitotic events, such as cyclin destruction (mitotic exit) and cytokinesis. This concerted action helps to maintain temporal co-ordination among mitotic events. It appears that the SCP is primarily activated by either a lack of occupancy or the absence of tension at kinetochores. Once triggered, the inhibitory circuit bifurcates, where one branch restrains the sister chromatid separation by inhibiting the E3 ligase APC(Cdc20) (anaphase-promoting complex activated by Cdc20) and the other impinges on the MEN (mitotic exit network). A large body of investigations has now led to the identification of the control elements, their targets and the functional coupling among them. Here we review the emerging regulatory network and discuss the remaining gaps in our understanding of this effective mechanochemical control system. PMID- 15521822 TI - Effects of deletions at the C-terminus of tobacco acetohydroxyacid synthase on the enzyme activity and cofactor binding. AB - AHAS (acetohydroxyacid synthase) catalyses the first committed step in the biosynthesis of branched-chain amino acids, such as valine, leucine and isoleucine. Owing to the unique presence of these biosynthetic pathways in plants and micro-organisms, AHAS has been widely investigated as an attractive target of several classes of herbicides. Recently, the crystal structure of the catalytic subunit of yeast AHAS has been resolved at 2.8 A (1 A=0.1 nm), showing that the active site is located at the dimer interface and is near the herbicide-binding site. In this structure, the existence of two disordered regions, a 'mobile loop' and a C-terminal 'lid', is worth notice. Although these regions contain the residues that are known to be important in substrate specificity and in herbicide resistance, they are poorly folded into any distinct secondary structure and are not within contact distance of the cofactors. In the present study, we have tried to demonstrate the role of these regions of tobacco AHAS by constructing variants with serial deletions, based on the structure of yeast AHAS. In contrast with the wild-type AHAS, the truncated mutant which removes the C-terminal lid, Delta630, and the internal deletion mutant without the mobile loop, Delta567-582, impaired the binding affinity for ThDP (thiamine diphosphate), and showed different elution profiles representing a monomeric form in gel-filtration chromatography. Our results suggest that these regions are involved in the binding/stabilization of the active dimer and ThDP binding. PMID- 15521823 TI - Demystifying borderline personality: critique of the concept and unorthodox reflections on its natural kinship with the bipolar spectrum. PMID- 15521824 TI - Attention deficit hyperactivity disorder and suicide: a review of possible associations. AB - OBJECTIVE: To review the evidence of a possible association between attention deficit hyperactivity disorder (ADHD) and suicide. DESIGN: We searched the electronic data bases: Medline, Psych LIT, between 1966 and March 2003 looking for articles on ADHD, attention deficit disorder, hyperactivity and suicide. RESULTS: An association of ADHD and completed suicide was found, especially for younger males. However, the evidence for any direct or independent link was modest with an overall suicide rate from long-term follow-up studies of ADHD of 0.63-0.78%. The estimated relative risk ratio, compared with US national suicide rates (males 5-24 years) is 2.91 (95% confidence interval 1.47-5.7, chi2 = 9.3, d.f. = 1, P = 0.002). ADHD appears to increase the risk of suicide in males via increasing severity of comorbid conditions, particularly conduct disorder (CD) and depression. CONCLUSION: Identification of those at risk, particularly males with comorbid ADHD, depression and CD, may represent a useful clinical means of reducing completed suicide. PMID- 15521825 TI - Axis II comorbidity of borderline personality disorder: description of 6-year course and prediction to time-to-remission. AB - OBJECTIVE: The purpose of this study was to compare the axis II comorbidity of 202 patients whose borderline personality disorder (BPD) remitted over 6 years of prospective follow-up to that of 88 whose BPD never remitted. METHOD: The axis II comorbidity of 290 patients meeting both DIB-R and DSM-III-R criteria for BPD was assessed at baseline using a semistructured interview of demonstrated reliability. Over 96% of surviving patients were reinterviewed about their co occurring axis II disorders blind to all previously collected information at three distinct follow-up waves: 2-, 4-, and 6-year follow-up. RESULTS: Both remitted and non-remitted borderline patients experienced declining rates of most types of axis II disorders over time. However, the rates of avoidant, dependent, and self-defeating personality disorders remained high among non-remitted borderline patients. Additionally, the absence of these three disorders was found to be significantly correlated with a borderline patient's likelihood-of remission and time-to-remission; self-defeating personality disorder by a factor of 4, dependent personality disorder by a factor of 3 1/2, and avoidant personality disorder by a factor of almost 2. CONCLUSION: The results of this study suggest that axis II disorders co-occur less commonly with BPD over time, particularly for remitted borderline patients. They also suggest that anxious cluster disorders are the axis II disorders which most impede symptomatic remission from BPD. PMID- 15521826 TI - Stressful life events as predictors of functioning: findings from the collaborative longitudinal personality disorders study. AB - OBJECTIVE: Although much attention has been given to the effects of adverse childhood experiences on the development of personality disorders (PDs), we know far less about how recent life events influence the ongoing course of functioning. We examined the extent to which PD subjects differ in rates of life events and the extent to which life events impact psychosocial functioning. METHOD: A total of 633 subjects were drawn from the Collaborative Longitudinal Personality Disorders Study (CLPS), a multi-site study of four personality disorders--schizotypal (STPD), borderline (BPD), avoidant (AVPD), obsessive compulsive (OCPD)--and a comparison group of major depressive disorders (MDD) without PD. RESULTS: Borderline personality disorder subjects reported significantly more total negative life events than other PDs or subjects with MDD. Negative events, especially interpersonal events, predicted decreased psychosocial functioning over time. CONCLUSION: Our findings indicate higher rates of negative events in subjects with more severe PDs and suggest that negative life events adversely impact multiple areas of psychosocial functioning. PMID- 15521827 TI - Culture, risk factors and suicide in rural China: a psychological autopsy case control study. AB - OBJECTIVE: Previous research on sociocultural factors for Chinese suicide have been basically limited to single case studies or qualitative research with ethnographic methodology. The current study examines the major risk factors and some cultural uniqueness related to Chinese rural suicide using a quantitative design. METHOD: This is a case control study with 66 completed suicides and 66 living controls obtained from psychological autopsy interviews in rural China. RESULTS: Both bivariate analyses and the multiple regression model have found that the Chinese rural suicide patterns are basically similar to those in most other cultures in the world: strong predictors of rural Chinese suicide are the psychopathological, psychological, and physical health variables, followed by social support and negative and stressful life events. Other significant correlates include lower education, poverty, religion, and family disputes. CONCLUSION: Culture has an important impact on suicide patterns in a society. PMID- 15521828 TI - The impact of suicidal ideation in preadolescence, adolescence, and young adulthood on psychosocial functioning and psychopathology in young adulthood. AB - OBJECTIVE: The long-term psychopathological outcome of suicidal ideation during adolescence in community samples is largely unknown. Data from the epidemiological and longitudinal Zurich Adolescent Psychology and Psychopathology Study (ZAPPS) allowed the testing of whether transient suicidal ideation in preadolescence or adolescence has the same impact on young adult psychopathology as enduring and concomitant suicidality in young adulthood. METHOD: Using various questionnaires on psychosocial functioning and the structured Composite International Diagnostic Interview (CIDI), the following five groups were compared: preadolescent suicidal ideation only, adolescent suicidal ideation only, young adult suicidal ideation only, enduring suicidality and young adult controls matched by age and gender. RESULTS: The young adult and the enduring risk group were most abnormal in various domains of psychosocial functioning and psychopathology. Whereas these groups differed from controls and the adolescent suicidal risk group, the differentiation from the preadolescent risk group was much weaker. CONCLUSION: There is a heightened risk of abnormal psychosocial and psychopathological functioning for young adult people with enduring and concomitant suicidal ideation and for those who experienced suicidal ideation in preadolescence. PMID- 15521829 TI - Rates of lifetime suicide attempt and rates of lifetime major depression in different ethnic groups in the United States. AB - OBJECTIVE: Rates of major depression and suicide vary across ethnic groups within the US. This also may be true of suicide attempts. METHOD: Data on lifetime suicidal behavior and major depression among Mexican American, Cuban American, and Puerto Rican adults who participated in the Hispanic Health and Nutrition Epidemiologic Survey were pooled with Epidemiological Catchment Area Study data for Blacks, Whites and Hispanics. RESULTS: Rates of major depression ranged from 9.3 (Puerto Ricans) to 3.24% (Cuban Americans). Puerto Ricans and whites had the highest rates of depression. Similarly, suicide attempt rates ranged from 9.1% for Puerto Ricans to 1.9% for Cuban Americans. Puerto Ricans had higher suicide attempt rates compared with other groups. CONCLUSION: This study underscores that there are differences between Hispanic ethnic groups. The impact of the migration process, socioeconomic status, and acculturation may underlie differences in major depression and suicide attempt rates across ethnic groups. PMID- 15521830 TI - Antidepressants and suicidal behaviour in unipolar depression. AB - OBJECTIVE: To compare the rates of suicidal behaviour during vs. after discontinuation of treatment with antidepressants, and to determine the comparative rates of suicidal behaviour for patients maintained on tricyclic (TCA) vs. selective serotonin reuptake inhibitor (SSRI) antidepressants. METHOD: Charts were reviewed for 521 patients with major depressive disorder and/or dysthymic disorder. Periods of active treatment or discontinuation with SSRIs or TCAs were determined. Rates of completed suicide, suicide attempts, and hospitalization for suicidality were analyzed. RESULTS: There was greater than a five-fold increase in risk for suicidal behaviour after discontinuation of antidepressant treatment (P < 0.0001). The rates of suicidal behavior during treatment with SSRIs or TCAs were similar. CONCLUSION: Suicidal behaviour in unipolar depressed patients treated with antidepressants increases substantially after medication discontinuation. This effect occurred in both patients who were maintained on SSRIs and TCAs. The findings support a possible protective effect on suicidal behaviour for both SSRIs and TCAs. PMID- 15521831 TI - Familial suicide behaviour: association with probands suicide attempt characteristics and 5-HTTLPR polymorphism. AB - OBJECTIVE: There is compelling evidence that a serotonergic dysfunction may play a major role in suicide behaviour and it has also been demonstrated that suicide is, at least partially, genetically determined. Thus, the serotonin-related genes are the major candidates. Previously a functional polymorphism in the promoter region of the serotonin transporter gene (5-HTTLPR) was identified and the presence of the short allele (S) was found to be associated with a lower level of expression of the gene and lower levels of 5-HT uptake when compared with the long allele (L). The purpose of this study was to evaluate the association between family suicide behaviour history and probands' suicide attempt (SA) history, SA characteristics and 5-HTTLPR genotype. METHOD: We genotyped 237 probands (major depressed or schizophrenic patients) and used a semistructured interview to determine probands' SA characteristics and first- and second-degree family suicidal behaviour. RESULTS: An association between suicidal family history and proband's SA but not with SA characteristics and probands genotype was found. CONCLUSION: Our results suggest that multiple biological and environmental factors underlie familial transmission of suicidal behaviour. PMID- 15521832 TI - The potential relationship between levels of perceived stress and subtypes of major depressive disorder (MDD). AB - OBJECTIVE: We wanted to explore whether major depressive disorder (MDD) subtypes (melancholic depression, atypical depression, double depression, and MDD with anger attacks) were related to levels of perceived stress, as measured by the Perceived Stress Scale (PSS). METHOD: Our sample [n = 298; female = 163 (55%); mean age 40.1 +/- 10.5 years] consisted of out-patients with MDD. The Structured Clinical Interview for DSM-III-R, the 17-item Hamilton Rating Scale for Depression, the Anger Attack Questionnaire, and the PSS were administered prior to initiating treatment. RESULTS: Depressed women had significantly higher levels of perceived stress (P = 0.02) than depressed men. Greater severity of depression at baseline was significantly related to higher levels of perceived stress (P < 0.0001). After adjusting for age, gender, and severity of depression at baseline, higher levels of perceived stress were significantly related to the presence of anger attacks (P < 0.0001; t = -4.103) as well as to atypical depression (P = 0.0013; t = 3.26). CONCLUSION: Out-patients with MDD who are more irritable and/or present with atypical features have higher levels of perceived stress, indicating a potential reactive component to their depression. PMID- 15521833 TI - Cognitive performance of male adolescents is lower than controls across psychiatric disorders: a population-based study. AB - OBJECTIVE: Psychiatric patients, as well as humans or experimental animals with brain lesions, often concurrently manifest behavioral deviations and subtle cognitive impairments. This study tested the hypothesis that as a group, adolescents suffering from psychiatric disorders score worse on cognitive tests compared with controls. METHOD: As part of the assessment for eligibility to serve in the military, the entire, unselected population of 16-17-year old male Israelis undergo cognitive testing and screening for psychopathology by the Draft Board. We retrieved the cognitive test scores of 19 075 adolescents who were assigned any psychiatric diagnosis, and compared them with the scores of 243 507 adolescents without psychiatric diagnoses. RESULTS: Mean test scores of cases were significantly poorer then controls for all diagnostic groups, except for eating disorders. Effect sizes ranged from 0.3 to 1.6. CONCLUSION: As group, adolescent males with psychiatric disorders manifest at least subtle impairments in cognitive functioning. PMID- 15521834 TI - Prediction of mortality at age 40 in Danish males at high and low risk for alcoholism. AB - OBJECTIVE: This prospective high-risk study examined the influence of father's alcoholism and other archival-generated measures on premature death. METHOD: Sons of alcoholic fathers (n = 223) and sons of non-alcoholic fathers (n = 106) have been studied from birth to age 40. Archival predictors of premature death included father's alcoholism, childhood developmental data, and diagnostic information obtained from the Psychiatric Register and alcoholism clinics. RESULTS: By age 40, 21 of the 329 subjects had died (6.4%), a rate that is more than two times greater than expected. Sons of alcoholic fathers were not more likely to die by age 40. Premature death was associated with physical immaturity at 1-year of age and psychiatric/alcoholism treatment. No significant interactions were found between risk and archival measures. CONCLUSION: Genetic vulnerability did not independently predict death at age 40. Death was associated with developmental immaturities and treatment for a psychiatric and/or substance abuse problem. PMID- 15521838 TI - The management of side-effects during therapy for hepatitis C. AB - The results of interferon and ribavirin combination therapy for chronic hepatitis C infection have substantially improved in recent years, such that the majority of patients in randomized-controlled trials now achieve a sustained virological response. However, adverse effects are commonplace, often disabling and may lead to interruption or cessation of therapy with subsequent loss of efficacy. Constitutional, neuropsychiatric and haematological reactions have proved particularly troublesome. In this review, we discuss these adverse effects in more detail and highlight recent advances in their management. PMID- 15521839 TI - Meta-analysis: a randomized trial of peginterferon plus ribavirin for the initial treatment of chronic hepatitis C genotype 4. AB - BACKGROUND: Guidelines for treatment of patients infected with hepatitis C virus genotype 4 are not available. AIM: To perform a meta-analysis of randomized controlled trials comparing peginterferon plus ribavirin with interferon plus ribavirin treatment in treatment-naive patients infected with chronic hepatitis C virus genotype 4. METHODS: The outcome measure was sustained virologic response. The measure of association employed was relative risk calculated by the random effect model, with heterogeneity, sensitivity and subgroup analyses. RESULTS: Of the 565 studies screened, six randomized controlled trials including 424 patients (peginterferon plus ribavirin 219, interferon plus ribavirin 205) were analysed. Duration of therapy was 1 year in all trials. Sustained virological response obtained with peginterferon plus ribavirin (55%) was significantly higher than with interferon plus ribavirin (30%) [relative risk, 1.71 (95% confidence interval, 1.15-2.56); P = 0.0088]. In the subgroup analyses, sustained virological response in trials using standard-dose ribavirin (1000 or 1200 mg/day) was 72% as against 45.8% in trials using low-dose ribavirin (800 mg/day) (P = 0.01). Further sub-group analyses for treatment duration, body weight, viral load and cirrhosis could not be performed because of lack of relevant data. CONCLUSION: Treatment-naive patients infected with hepatitis C virus genotype 4 should be treated with peginterferon plus standard-dose ribavirin for 1 year, with an expected sustained virological response rate of 72%. PMID- 15521840 TI - Personal view: cost and benefit of medical rituals in gastroenterology. AB - BACKGROUND: Unable to resolve a medical problem, gastroenterologists occasionally choose an ineffectual intervention instead. The elusive path to effectual management becomes substituted with an ineffectual but readily available medical ritual. The term 'ritual' refers to the utilization of an ineffectual intervention with little chances of achieving a medically relevant goal. AIM: The aim of the article is to analyse the cost-benefit relationship of rituals in gastroenterology and the reasons why gastroenterologists utilize them. METHODS: Ritualistic disease management is described in terms of medical decision and threshold analysis. RESULTS: If the perceived benefit of a ritual exceeds its cost, the ineffectual path becomes more attractive than expectant management or doing nothing. To engage in an ineffectual intervention, the threshold probability for success needs to exceed its cost-benefit difference divided by the benefit associated with success. In choosing an ineffectual intervention, doctors tend to underestimate the true costs of a ritual and overestimate its probability of success. The availability of an effectual path leads to more stringent conditions for the benefit associated with an ineffectual path. CONCLUSIONS: A better understanding of their economical underpinnings should lead to more restricted utilization of rituals in gastroenterology, especially, in situations where they drain scarce resources disproportionately to their perceived benefit. PMID- 15521841 TI - A prospective, randomized-controlled pilot study of ursodeoxycholic acid combined with mycophenolate mofetil in the treatment of primary sclerosing cholangitis. AB - BACKGROUND: Ursodeoxycholic acid has been ineffective in the treatment of primary sclerosing cholangitis. Because the pathogenesis of primary sclerosing cholangitis is related to immune destruction of bile duct epithelium, several immune suppressive agents have been evaluated. Mycophenolate mofetil is a potent immunosuppressant that is now widely used in organ transplantation. AIM: In this pilot study to determine if mycophenolate mofetil when combined with ursodeoxycholic acid could prevent evidence of clinical progression and improve the biochemical, histological and/or cholangiographic features of primary sclerosing cholangitis compared with patients treated with ursodeoxycholic acid alone. METHODS: Twenty-five patients with well-defined primary sclerosing cholangitis were randomized to ursodeoxycholic acid (13-15 mg/kg/day) with or without mycophenolate mofetil (1000 mg b.d.). Cholangiography and liver biopsy were performed at study entry and after 2 years of treatment. Symptoms, clinical features of liver disease and biochemical tests were monitored at 3-month intervals. RESULTS: The mean age 44 years, 58% male, 84% Caucasian and 64% had ulcerative colitis. After 2 years, there were no differences in laboratory values, histological stage or cholangiograms between patients treated with ursodeoxycholic acid alone and those treated with mycophenolate mofetil + ursodeoxycholic acid. CONCLUSIONS: Mycophenolate mofetil combined with ursodeoxycholic acid does not appear to provide additional benefit compared with standard doses of ursodeoxycholic acid alone in the treatment of primary sclerosing cholangitis. PMID- 15521842 TI - Rapid hip bone loss in active Crohn's disease patients receiving short-term corticosteroid therapy. AB - BACKGROUND: Uncertainty over whether corticosteroids cause bone loss in patients with Crohn's disease may reflect their short, intermittent use. AIM: We investigated whether a 2-month course of prednisolone is associated with detectable bone loss. METHODS: Fifteen patients with active Crohn's disease and 19 controls with inactive Crohn's disease were recruited. Bone mineral density of the lumbar spine and hip was measured at baseline and 2 and 8 months. RESULTS: At 2 months, significant bone loss was found in patients with active disease (femoral neck -2.7%, P < 0.002; Ward's triangle -3.9%, P < 0.01). Although bone mineral density was still lower at 8 months, these differences were no longer significant (-1.3% and -3.4%, femoral neck and Ward's triangle, respectively). No significant change in hip bone mineral density was observed in controls. Previous corticosteroid use was not significantly associated with baseline bone mineral density, although significant independent associations were observed between weight, site of disease and lumbar spine bone mineral density, and between dietary calcium deficiency and femoral neck and Ward's triangle bone mineral density. CONCLUSION: Significant bone loss at the hip can be detected in patients receiving corticosteroid treatment for 2 months for active Crohn's disease ; however, it remains unclear whether this is because of disease activity or its treatment. This rapid bone loss may represent a risk factor for fracture and justify bone protective therapy. PMID- 15521843 TI - Behaviour of the bowel wall during the first year after surgery is a strong predictor of symptomatic recurrence of Crohn's disease: a prospective study. AB - BACKGROUND: Recurrences after surgery for Crohn's disease are frequent and unpredictable. To date, there is little agreement as to which factors increase a patient risk of early recurrence. AIM: To assess whether the post-operative behaviour of diseased bowel walls, as determined by ultrasound, may be a useful predictor of relapse. METHODS: A total of 127 Crohn's disease patients were monitored after surgery by means of bowel ultrasound as well as by clinical and laboratory evaluations for a median follow-up of 41.0 months. Bowel wall thickness of diseased loops measured at ultrasound during follow-up was compared with the presurgery values. Multivariable survival analysis was performed to elucidate predictors of early post-operative recurrence. Receiver operating characteristic curves were also constructed taking into account bowel wall thickness for selecting Crohn's disease patients with high risk of clinical/surgical recurrence. RESULTS: The estimated 5 years survival probability of symptomatic Crohn's disease recurrence were 90% and 33%, respectively for unchanged/worsened bowel wall thickness vs. improved bowel wall thickness at 12 months from surgery. The hazard ratio for unchanged/worsened bowel wall thickness at 12 months was 8.9 (95% CI: 3.4-23.2). Receiver operating characteristic curve identified a bowel wall thickness > 6.0 mm at 12 months from surgery as directly associated with the risk of having a Crohn's disease recurrence (hazard ratio was 6.5, 95% CI: 2.8-15.4). CONCLUSIONS: Systematic ultrasound follow-up of diseased bowel walls after conservative surgery allows the early identification of patients at high risk of clinical/surgical recurrence. PMID- 15521844 TI - Sleep dysfunction in patients with gastro-oesophageal reflux disease: prevalence and response to GERD therapy, a pilot study. AB - BACKGROUND: There is little information on the prevalence of pathological sleep disorders in patients with gastro-oesophageal reflux disease and whether pharmacological treatment of patients with gastro-oesophageal reflux disease will lead to improvement in sleep. AIMS: This pilot study determined the prevalence of sleep disorder in patients with erosive gastro-oesophageal reflux disease, correlated subjective (questionnaire) and objective (actigraphy - a watch worn on the wrist that monitors motion to help differentiate sleep from awake states) assessment of sleep dysfunction and determined whether therapeutic resolution of oesophageal symptoms was associated with an improvement in sleep. METHODS: Eighteen patients with erosive gastro-oesophageal reflux disease received esomeprazole 40 mg once daily for 8 weeks. Assessments at 0, 4 and 8 weeks included: Gastrointestinal Symptoms Rating Scale, Pittsburgh Sleep Quality Index questionnaire and ambulatory wrist actigraphy. RESULTS: Unrecognized sleep disturbance occurred in 81% of this cohort of patients with gastro-oesophageal reflux disease and erosive oesophagitis. Median reflux syndrome score (heartburn and acid regurgitation) on Gastrointestinal Symptoms Rating Scale decreased from 2 at baseline to 0 at weeks 4 and 8 (P 2 or <0.5) including provision of infertility treatment to lesbian (46.9% vs 16.7%) and homosexual couples (28.4% vs 11.4%), the opportunity for homosexual couples to use surrogate mothers (30.6% vs 15.2%) and limitations in the number of infertility treatment cycles (28.4% vs 61.4%). For 11 questions, we recorded minor, but statistically significant, differences. In the prioritisation questions, the women set the order according to their own interests, probably because the women were at fertile age and they had or would like to have a child. Maternity services and screening for cancer in women (Papanicolaou's test and mammography) were at the top of the list. CONCLUSION: These results reflect a split attitude that was influenced by the wish of infertile women to help childless couples and to be able to recruit suitable sperm/oocyte donors. Parous women were motivated by their concern for children's rights. PMID- 15521868 TI - The predictive value of the "Hull & Rutherford" classification for tubal damage. AB - OBJECTIVE: This study explores the predictive value for live birth following tubal reconstructive surgery of the "Hull & Rutherford" (H&R) classification system. DESIGN: Retrospective cohort study. SETTING: Tertiary infertility referral service, University of Bristol. POPULATION: Infertile women younger than 40 years with tubal damage undergoing tubal surgery. METHODS: Women (n= 192) were grouped according to three severity grades of disease based on the H&R classification. Essentially, the main features of grade I tubal damage were filmy adhesions, whereas grades II and III referred to unilateral severe damage and bilateral severe damage, respectively. Standard surgical techniques were employed. Pregnancy and live birth rates were calculated and compared using time specific univariate Kaplan-Maier curves and multivariate Cox's regression analysis. MAIN OUTCOME MEASURES: Pregnancy, ectopic and live birth within three years of surgery. RESULTS: A significant trend towards higher ectopic pregnancy rates (P < 0.001) with increasing severity of tubal damage was noted, but not miscarriage rates. Univariate analysis revealed significant differences in the live birth rates of 69%, 48% and 9% for grades I, II and III, respectively. Multivariate analysis (controlling for age, duration of and primary infertility) confirmed these differences to be significant with risk ratios of 13.7 (95% CI: 4.49-41.9) and 6.54 (95% CI: 2.48-17.24) for grades I and II disease, respectively, compared with grade III disease, used as the reference. CONCLUSIONS: The H&R classification is a simple classification system that is able to distinguish women into three distinct groups giving a favourable, fair and poor prognosis for live birth following tubal surgery. PMID- 15521869 TI - Ki-67, Bcl-2 and p53 expression in endometrial polyps and in the normal endometrium during the menstrual cycle. AB - OBJECTIVE: To determine the presence of proteins related to proliferation (Ki-67) and apoptosis (Bcl-2, p53) in endometrial polyps and normal endometrium during the menstrual cycle. DESIGN: Retrospective study using paraffin embedded tissue. SETTING: Hospital affiliated to the university. POPULATION: Premenopausal patients with endometrial polyps. METHODS: Seventy-eight premenopausal patients in different phases of the menstrual cycle were submitted to polypectomy using the Bettocchi hysteroscope. Immunohistochemistry was used to detect the expression of these proteins in endometrial polyps. One hundred and eighteen normal endometrial biopsies were used as controls. MAIN OUTCOME MEASURES: Detection of Bcl-2 and Ki-67 expression by immunohistochemistry. RESULTS: In endometrial polyps, Ki-67, p53 and Bcl-2 expression was detected with more frequency during the proliferative than during the luteal phase of the cycle. Similar findings were observed in the normal endometrium. CONCLUSION: Endometrial polyps undergo cyclic changes in the expression of their proteins related to proliferation and apoptosis during the menstrual cycle, similar to those of the cycling endometrium. PMID- 15521870 TI - Inhibin B and anti-Mullerian hormone: markers of ovarian response in IVF/ICSI patients? AB - OBJECTIVE: The objective of this study was to investigate whether follicle stimulating hormone (FSH), anti-Mullerian hormone (AMH) and inhibin B could be useful in predicting the ovarian response to gonadotrophin stimulation in assisted reproduction patients who are considered to be poor responders. DESIGN: Prospective study. SETTING: Fertility unit. SAMPLE: Blood samples were collected on day five or six in the early follicular phase of an untreated menstrual cycle. Samples were collected from 69 patients. METHODS: Serum samples were assayed for FSH, AMH and inhibin B using commercial immunoassay kits. MAIN OUTCOME MEASURES: Response to gonadotrophin stimulation and number of eggs collected. RESULTS: Among the 69 patients, 52 patients completed an IVF cycle and 17 patients had to cancel the cycle because of poor ovarian response to gonadotrophin stimulation. Mean FSH levels were significantly higher (P < 0.05) in the cancelled group (10.69 +/- 2.27 mIU/mL) compared with the cycle-completed group (7.89 +/- 0.78 mIU/mL). Mean AMH levels were significantly lower (P < 0.01) in the cancelled group (0.175 +/- 0.04 ng/mL) compared with the cycle-completed group (1.13 +/- 0.2 ng/mL). Mean inhibin B levels were significantly lower (P < 0.001) in the cancelled group (70 +/- 12.79 pg/mL) compared with the completed group (126.9 +/- 8.8 pg/mL). Predictive statistics show that AMH is the best single marker and that the combination of FSH, AMH and inhibin B is modestly better than the single marker. Linear regression analysis in the cycle completed patients shows that although FSH (r= 0.25, P < 0.05) and inhibin B (r= 0.35, P < 0.05) have a significant linear association with the number of eggs collected, AMH has the greatest association (r= 0.69, P < 0.001) with the number of eggs collected among the parameters measured. CONCLUSION: In this particular group of IVF patients, AMH is the best single marker of ovarian response to gonadotrophin stimulation. The combined markers modestly improved the prediction. PMID- 15521871 TI - Is induced abortion a contributing factor to tubal infertility in Mexico? Evidence from a case-control study. AB - OBJECTIVE: To determine the contribution of induced abortion to tubal infertility in Mexico. DESIGN: Population- and hospital-based case-control study. SETTING: Tertiary hospitals in Mexico City. SAMPLE: Women between 20 and 40 years with infertility and controls of the same age: 251 cases, 502 hospital controls, 502 neighbourhood controls. METHODS: A case-control study was conducted in four tertiary hospitals in Mexico City with 251 cases and 1004 controls (two hospital and two neighbourhood controls per case, matched by age [+/-2 years]). Cases were infertile women, aged 20-40, with tubal occlusion confirmed by laparoscopy. Controls were fertile women, who had carried a pregnancy to term within the last two years. Participants completed a previously validated questionnaire asking about reproductive history and induced abortion. RESULTS: Our study did not show an association between induced abortion and tubal infertility among women that did not relate both events (cases vs hospital controls: OR = 1.57, 95% CI: 0.29 8.65; cases vs neighbourhood controls: OR = 0.82, 95% CI: 0.07-8.99) using conditional logistic models adjusting by marital status, number of pregnancies, age at first sexual intercourse and history of pelvic inflammatory disease. In contrast, early age at sexual debut and history of pelvic inflammatory disease significantly increased the risk of tubal infertility. CONCLUSIONS: In Mexico, the lack of association between induced abortion and tubal damage causing infertility observed in this population might be explained by a shift toward "safer" abortion practices. PMID- 15521872 TI - An evaluation of economics and acceptability of screening for Chlamydia trachomatis infection, in women attending antenatal, abortion, colposcopy and family planning clinics in Scotland, UK. AB - OBJECTIVE: The aims of this study were to determine cost effectiveness of screening for Chlamydia trachomatis in hospital-based antenatal and gynaecology clinics, and community-based family planning clinics. Additionally, women's views of screening were determined in the hospital-based clinics. DESIGN: Cost effectiveness based on decision model. Model probabilities were generated for a hypothetical sample of 250 women in each age group in each setting, based on prevalence studies, published data and expert opinion. A prospective observational study was used to generate data on prevalence and acceptability. SETTING: Antenatal, gynaecology and family planning clinics in Aberdeen, Edinburgh and Glasgow. SAMPLE: Prevalence was estimated in 2817 women. Acceptability was determined in 484 women. METHODS: An economic evaluation was performed using prevalence data from this and a previous study, and using outcome data from the literature and observational work. Incremental cost effectiveness ratios were estimated for each age group and clinical setting. Sensitivity analyses were performed to determine the robustness of incremental cost effectiveness ratios to changes in the incidence of long term sequelae and costs. The prevalence of infection was determined by nucleic acid amplification of urine samples or endocervical swabs. Knowledge of C. trachomatis and women's views of screening were determined using structured questionnaires. MAIN OUTCOME MEASURES: Direct health service costs of screening, incidence and costs associated with adverse sequelae, women's views of screening and prevalence of infection. RESULTS: The estimated cost of screening 250 women in each age group in each the four sample populations (total population of 3750) is 49,367 UK pounds, while preventing 64 major sequelae. This represents a net cost of 771.36 UK pounds in preventing one major sequela. Selective screening of all women under 20 years and all patients attending abortion clinics were shown to be the most cost effective strategies. These results were relatively insensitive to changes in estimated parameters, such as uptake rate, probabilities and unit costs of all major sequelae averted. Prevalence (95% CI) of infection in the highest risk groups (those aged under 20 in both antenatal and abortion clinics) was 12.1% (8.6-16.7) and 12.7% (7.3-21.2), respectively. The majority (>95%) of women agreed with a policy of regular screening for C. trachomatis, and screening in the settings employed in this study was largely acceptable. CONCLUSIONS: A single episode of screening for C. trachomatis does not result in net cost savings. Currently recommended strategies of screening for C. trachomatis in women under 25 years of age in abortion clinics are supported by our data on prevalence and acceptability. These data also suggest that hospital-based screening strategies should be further extended to include younger women attending antenatal clinics and all women of reproductive age attending colposcopy clinics. PMID- 15521873 TI - Current practice for the laparoscopic diagnosis and treatment of endometriosis: a national questionnaire survey of consultant gynaecologists in UK. AB - OBJECTIVE: To determine current practice regarding laparoscopic diagnosis and treatment of endometriosis. DESIGN: A prospective questionnaire survey. SETTING: The United Kingdom. POPULATION: All 1411 UK consultant gynaecologists identified from a Royal College of Obstetricians and Gynaecologists database. METHODS: A postal questionnaire was sent to all consultants with reply paid envelopes. A postal reminder was sent three months following the initial questionnaire. MAIN OUTCOME MEASURE: Current practice for the laparoscopic diagnosis and treatment of endometriosis and willingness to participate in a randomised trial. RESULTS: The response rate was 66% (893/1411). Diagnostic laparoscopy was performed by 87% (772/893) of respondents. Seventy-six percent of these (58/772) were confident to visually diagnose endometriosis and 6% (47/772) routinely verified the diagnosis histologically. Laparoscopic surgery was routinely undertaken by 41% (318/772) of respondents. Ablative therapy was the most frequently employed technique utilised [620/653 (95%)] and electrodiathermy was the most popular energy modality (80%). Among respondents expressing a preference, excision of disease was believed to be more effective, but less safe compared with ablation. One-third of respondents (273/893) were willing to enter patients into a randomised controlled trial to compare laparoscopic treatments for pelvic pain associated with endometriosis. CONCLUSION: Laparoscopic surgery for endometriosis associated with pelvic pain is routinely undertaken by a large number of UK consultant gynaecologists, but techniques used and beliefs about efficacy vary. In view of this division of opinion regarding the relative roles of laparoscopic treatment methods, a randomised trial comparing the efficacy and safety of these methods is urgently needed. PMID- 15521874 TI - Bed rest versus free mobilisation following embryo transfer: a prospective randomised study. AB - OBJECTIVE: To evaluate the efficacy of two clinical methods of post-embryo transfer protocols in patients undergoing in vitro fertilisation. DESIGN: Prospective, randomised clinical trial. SETTING: Hospital-based clinic for reproductive medicine. SAMPLE: Women under 40 years of age who were undergoing in vitro fertilisation with GnRH pituitary down-regulation and controlled ovarian hyperstimulation. METHODS: Patients were randomised to rest for either 1 or 24 hours after embryo transfer. MAIN OUTCOME MEASURE: Clinical pregnancy per cycle rate (the percentage of cycles started that demonstrated a live fetus on ultrasound examination performed at six or seven weeks of gestation). RESULTS: The clinical pregnancy rates were 21.5% for the 1-hour and 18.2% for the 24-hour post-embryo transfer groups. The implantation rate per embryo was significantly higher in the 1-hour group (14.4%) than in the 24-hour group (9%). CONCLUSION: One-hour and 24-hour rest post-embryo transfer result in comparable rates of clinical pregnancy. However, 24-hour rest results in reduced implantation rate per embryo. PMID- 15521875 TI - Low dose recombinant FSH treatment may reduce multiple gestations caused by controlled ovarian hyperstimulation and intrauterine insemination. AB - OBJECTIVE: To evaluate the rate of multiple pregnancies in intrauterine insemination cycles stimulated with a minimal dose of recombinant follicle stimulating hormone (rec-FSH). DESIGN: Retrospective study. SETTING: University Medical Center. POPULATION: A total of 1256 patients underwent 3219 consequent intrauterine insemination cycles with minimal ovarian stimulation. METHODS: Patients received 50 or 75 IU of rec-FSH from day four to day seven. The dose was adjusted according to oestradiol (E(2)) levels in order to achieve a maximum of two follicles on the day of hCG administration. MAIN OUTCOME MEASURES: Peak E(2) levels, the number of follicles >15 mm and pregnancy rates were calculated. The predictive value of E(2) levels for multiple gestations was also estimated. RESULTS: Of 3219 cycles, 334 resulted in pregnancies (10%). Of these, 238 (91%) were singletons, 28 (8%) twins and 1 (0.3%) was a triplet. The cumulative overall pregnancy rate was 43%. Patients over 40 years old had a significantly lower pregnancy rate per cycle and overall live birth rate (P < 0.05). Most pregnancies (83%) occurred during the first three cycles. Pregnancy rates per cycle varied from 8% for tubal factor to 14% for anovulation infertility. CONCLUSIONS: Minimal FSH stimulation in intrauterine insemination cycles may reduce the rates of twins and high order multiple pregnancies without affecting overall pregnancy rates. PMID- 15521876 TI - The conservative management of interstitial pregnancy. AB - OBJECTIVES: To evaluate the effectiveness of systemic methotrexate in the treatment of interstitial pregnancy. DESIGN: Prospective observational study. SETTING: An Early Pregnancy Assessment Unit in a London teaching hospital. SAMPLE: Twenty consecutive women diagnosed with an interstitial pregnancy. METHODS: Women were diagnosed with an interstitial pregnancy based on transvaginal ultrasound findings. Single dose, intramuscular methotrexate was administered on day 0. A second dose of methotrexate was given if the beta-hCG levels had not fallen by 15% between days four and seven. Weekly follow up continued until the serum beta-hCG < 5 IU. MAIN OUTCOME MEASURE: The resolution of serum beta-hCG levels without the need for surgical intervention. RESULTS: Two hundred and ninety-three ectopic gestations were diagnosed over a 42-month period. Twenty of these were interstitial in nature, with a median initial serum beta-hCG of 6452 IU. Of the 20 interstitial pregnancies, 17 cases received systemic methotrexate. Sixteen were treated successfully (94%), including all of the four cases with fetal heart activity present. A second methotrexate dose was given to six patients. Two cases were managed expectantly. Two cases underwent laparotomy and cornual resection: one elected for surgical management at the outset and one as a result of suspected ectopic rupture after two doses of methotrexate. There were no other complications. CONCLUSIONS: Systemic methotrexate is a safe and highly effective treatment for interstitial pregnancy. Surgery can be avoided in the majority of women with this condition. Early recognition of the cornual pregnancy with transvaginal ultrasound is essential. PMID- 15521877 TI - Should cryopreserved epididymal or testicular sperm be recovered from obstructive azoospermic men for ICSI? AB - OBJECTIVE: To determine the effect of the anatomical site of sperm recovery on intracytoplasmic sperm injection (ICSI) embryo implantation, pregnancy and live birth rates in couples with isolated obstructive azoospermia as the sole cause of infertility. DESIGN: Controlled, single centre, retrospective clinical study. SETTING: University Hospital, Centre for Reproductive Medicine. SAMPLE: One hundred and fifty-one cycles of ICSI were performed, using surgically recovered sperm, between August 1996 and March 2002. METHODS: The outcome of ICSI, with surgically recovered sperm, was compared between epididymal (Group E) and testicular (Group T) derived sperm. Inclusion was limited to couples undergoing their first treatment cycle, where female age was < or =39 years and a minimum of five oocytes were available for injection. Women with a history of ovarian surgery, ultrasonic evidence of polycystic ovaries, uterine anomalies or hydrosalpinx were excluded. MAIN OUTCOME MEASURES: Clinical pregnancy, implantation and live birth rate. RESULTS: Forty-two of 151 cycles met the strict inclusion criteria. Groups E and T were comparable with respect to age, basal serum FSH, ovarian response; number of oocytes injected and number of embryos available and transferred. No difference existed between Groups E and T in implantation, clinical pregnancy or live birth rate (28.8% vs 25.8%, 42.9% vs 42.9% and 39.3% vs 42.9%, respectively). CONCLUSIONS: Cryopreserved epididymal and testicular sperm, from men with obstructive azoospermia, appear equally effective in ICSI. Epididymal recovery should remain the method of first choice for obstructive azoospermic men but further study of sperm DNA damage rates in different testicular sites is required. PMID- 15521878 TI - Adverse maternal outcomes in multifetal pregnancies. AB - In this retrospective cohort of 165,188 singleton pregnancies and 44,674 multiple fetal pregnancies in Canada from 1984 to 2000, we compared the incidence of maternal complications. Multiple gestation pregnancies were associated with significant increases in cardiac morbidity, haematologic morbidity, amniotic fluid embolus, pre-eclampsia, gestational diabetes, postpartum haemorrhage, prolonged hospital stay, the need for obstetric intervention, hysterectomy and blood transfusion. Multiple gestation pregnancies are associated with an increased risk of morbidity for the mother. This should be taken into consideration in antenatal care of these women. PMID- 15521879 TI - Management of ovarian hyperstimulation due to follicle-stimulating hormone secreting gonadotroph adenoma. PMID- 15521880 TI - Fatal mesenteric artery thrombus following oocyte retrieval. PMID- 15521881 TI - The global state of prostate cancer: epidemiology and screening in the second millennium. PMID- 15521882 TI - The global state of prostate cancer: new diagnostic tools, minimal requirements for diagnosis and staging, and guidelines in the second millennium. PMID- 15521883 TI - Advanced prostate cancer treatment guidelines: a global perspective; trends of hormone therapy in Japan. PMID- 15521884 TI - Advanced prostate cancer treatment guidelines: European perspective. PMID- 15521885 TI - Advanced prostate cancer treatment guidelines: a United States perspective. PMID- 15521886 TI - Biochemical failure: role for early hormonal therapy. PMID- 15521887 TI - Assessment and management of high-risk patients. PMID- 15521888 TI - Hormonal treatment alone for locally advanced prostate cancer. PMID- 15521889 TI - Management of hormone-refractory disease. PMID- 15521891 TI - Individualizing drug therapy, and 'men behaving badly'. PMID- 15521893 TI - Effect of oral linezolid on the pressor response to intravenous tyramine. AB - AIMS: To investigate the effect of monoamine oxidase A inhibition from a single oral dose of linezolid on the pressor response to intravenous (i.v.) tyramine, using positive and negative controls to validate the methodology. METHODS: This placebo-controlled, three-period crossover study was conducted in 12 healthy male volunteers. Each volunteer received either one oral dose of moclobemide (300 mg), linezolid (600 mg), or placebo tablet followed by an i.v. tyramine pressor test until an increase in systolic blood pressure of at least 30 mmHg above baseline occurred. Each study day was separated by a 7-day washout period. The dose of tyramine required to raise the blood pressure by 30 mmHg (TYR30) was calculated for each oral treatment by linear interpolation between log-transformed doses of i.v. tyramine. The influence of body mass index (BMI) on TYR30 was also investigated. RESULTS: The tyramine sensitivity factor (ratio of the geometric least square mean TYR30 for placebo and active oral treatment) was 1.8 [90% confidence interval (CI) 1.6, 2.0, P < 0.0001] for linezolid and 2.1 (90% CI 1.8, 2.4, P < 0.0001) for the positive control moclobemide. BMI had a statistically significant effect on TYR30. CONCLUSIONS: There was a significant difference in the pressor response to i.v. tyramine between linezolid and placebo. Moclobemide (positive control) and linezolid have a similar pressor response to i.v. tyramine. The statistically significant effect of BMI on TYR30 underlines the advantage of within-individual comparisons of treatments in order to reduce variability and provide more accurate treatment estimates. PMID- 15521892 TI - Drug development and use in the elderly: search for the right dose and dosing regimen (Parts I and II). PMID- 15521894 TI - Effect of itraconazole on the pharmacokinetics and pharmacodynamics of a single oral dose of brotizolam. AB - AIMS: To assess the effect of itraconazole, a potent inhibitor of cytochrome P450 (CYP)3A4, on the single oral dose pharmacokinetics and pharmacodynamics of brotizolam. METHODS: In this randomized, double-blind, cross-over trial 10 healthy male subjects received either itraconazole 200 mg or matched placebo once daily for 4 days. On day 4, a single 0.5 mg dose of brotizolam was administered orally. Plasma concentrations of brotizolam were followed up to 24 h, together with assessment of psychomotor function measured by the digit symbol substitution test (DSST), visual analogue scales and UKU side-effect rating scale. RESULTS: Itraconazole significantly (P < 0.001) decreased the apparent oral clearance (CL/F) (16.47 +/- 4.3 vs 3.91 +/- 2.1), increased the area under the concentration-time curves (AUC) from 0 h to 24 h (28.37 +/- 10.8 vs 68.71 +/- 24.1 ng ml h(-1)), and prolonged the elimination half-life (4.56 +/- 1.4 vs 23.27 +/- 10.3 h) of brotizolam. The AUC(0,24 h) of the DSST (P < 0.001) and the item 'sleepiness' of UKU (P < 0.05) were significantly decreased. CONCLUSIONS: Itraconazole increases plasma concentrations of brotizolam probably via its inhibitory effect on CYP3A4 brotizolam metabolism. PMID- 15521895 TI - Internet based clinical trial protocols -- as applied to a study of warfarin pharmacogenetics. AB - AIMS: To describe and evaluate the use of an Internet-based study protocol in a multicentre study of genetic risk factors in anticoagulant treatment. METHODS: A web-based study protocol, similar to existing anticoagulation medical record systems, was developed for entry of clinical data. It was also supplied with a separate interface for study monitoring. Measures were taken to assure the confidentiality of transferred data. In addition, software modifications were made to enable automated transfer of clinical data from an existing medical record system to the study database. RESULTS: The system has been in use since March 2002, and at present 39 centres have included 909 patients with a dropout rate of 2.8%. The need for education of participating clinicians has been satisfactorily provided for by means of written instructions and telephone support. CONCLUSIONS: Our study demonstrates the usability of Internet-based data acquisition techniques in a full-scale multicentre clinical trial. The main advantages of such a protocol are automated data validation and standardization, fast data transfer independent of geographical distance, user feedback, synchronization of protocol updates and automatic data formatting facilitating statistical analyses. Safety and accessibility are possibly cumbersome areas and should be addressed duly. PMID- 15521896 TI - Adsorption of oxacalcitriol by polysulphone haemodialyser in patients with secondary hyperparathyroidism. AB - AIMS: This study was undertaken to evaluate removal of 22-oxacalcitriol (OCT), an active and intravenously used vitamin D3 analogue with less calcaemic activity, by polysulphone haemodialyser in vivo and in vitro. We further compared the pharmacodynamic efficacy [suppression of intact parathyroid hormone (iPTH)] when given intravenously either during or at the end of the haemodialysis. METHODS: (i) Drug clearance by the polysulphone dialyser was measured during a single continuous infusion (5 microg) for 30 min into the arterial side of the dialyser in end-stage renal failure patients with secondary hyperparathyroidism (n = 7). (ii) The drug adsorption by the hollowfibre membrane during incubation for 30 min was measured in vitro. (iii) To evaluate efficacy, the drug was given (i.v. bolus) during or at the end of haemodialysis for 4 weeks in a cross-over fashion with a washout period of 8 weeks (n = 9). Serum Ca(2+), phosphate (P) and iPTH concentrations just before the initiation of the dialysis were monitored every week. RESULTS: (i) OCT was significantly cleared by the polysulphone haemodialyser, but the clearance declined in a time-dependent manner to approach zero at 30 min. Arterial (at the place between the drug infusion site and the haemodialyser column) drug concentrations did not change during the infusion (mean = 2064 +/- 233 pg ml(-1)). Venous (just after the dialyser) drug concentrations at 10 min after the infusion were significantly lower than those of the arterial side (mean = 784 +/- 84 pg ml(-1)); however, they increased with time and reached those of the arterial side at 30 min. (ii) In vitro, OCT was adsorbed by the membrane. The amount of adsorption was concentration-dependent and was lower in the presence of human serum (55 +/- 4% without and 23 +/- 4% with serum at 600 pg ml(-1) of OCT). (iii) Although serum Ca(2+) and P increased and iPTH decreased by both treatment regimens (i.e. OCT administered either during or at the end of haemodialysis), these changes did not significantly differ. Mean differences (and 95% confidence interval) of Ca(2+), P, and iPTH at the end of the trial were 0.03 (-0.04, 0.09) mm, 0.41 (-0.43, 1.26) mg dl(-1) and 38 (-42, 88) pg ml(-1), respectively. CONCLUSION: OCT is adsorbed by polysulphone dialyser in vitro and in vivo. However, the pharmacodynamic effectiveness was largely independent of the administration regimen of OCT given either during or at the end of haemodialysis. PMID- 15521897 TI - Influence of socioeconomic status on the quality of prescribing in the elderly -- a population based study. AB - AIMS: To compare the quantity and quality of prescribing, using prescribing indicators, between the relatively deprived and the relatively affluent patients over 70 years old in primary care. METHODS: We examined the General Medical Services (GMS) scheme prescribing data for the Eastern region in Ireland for all patients aged 70 years or more (n = 95 055) during July 2001-December 2002. Prescribing indicators applied to the prescription data to evaluate drugs prescribed to each patient were classified as: (1) descriptive, e.g. number of drug items/patient; (2) indicators assessing potentially harmful prescribing; (3) evidence-based indicators, e.g. secondary prevention therapy in those with ischaemic heart disease (IHD). Results are expressed as odds ratio (OR) and 95% confidence intervals for comparison of prescribing between the relatively deprived and affluent cohorts, adjusted for age and gender (CI) using logistic regression analysis. RESULTS: The relatively deprived cohort was more likely to receive a greater number of drugs (5.2 vs. 4.5, P < 0.0001), more generic products (P = 0.01) and be exposed to major polypharmacy (> or = five drugs) compared with monotherapy (OR = 1.58; 95% CI = 1.52,1.64, P < 0.0001). They were more likely to receive potentially harmful drugs such as cerebral vasodilators (OR = 1.52; 1.38,1.69), long-acting sulphonylurea (OR = 1.43; 1.20,1.72), potentially interacting agents such as angiotensin converting enzyme (ACE) inhibitor and potassium sparing diuretic (OR = 1.78; 1.48,2.15). In terms of evidence-based prescribing, the relatively deprived cohort with IHD was less likely to receive secondary prevention therapies such as statins (OR = 0.82; 0.74, 0.90) and beta blockers (OR = 0.85; 0.77,0.93). CONCLUSIONS: These results show that suboptimal prescribing is more prevalent among the relatively deprived cohort and suggests the differences observed in the quality of prescribing between these patient groups may be related to their socioeconomic status. PMID- 15521898 TI - Evaluation of the clinical value of pharmacists' modifications of prescription errors. AB - AIMS: Our objective was to examine the clinical value of pharmacists' interventions to correct prescription errors. METHODS: In this study, we reviewed a random sample of prescriptions that had been modified in pharmacies. These prescriptions were collected on one predetermined day between 25th February and 12th March 1999 from 141 Dutch community pharmacies. Each prescription modification was evaluated by a panel of reviewers, including representatives of five groups of health care professionals. After generally rating each modification as positive, negative, or neutral, the reviewers assessed its outcome (in terms of prevention of an adverse drug reaction [ADR], an improvement in effectiveness, both, or other), the probability and importance of improvements in effectiveness and/or the probability and seriousness of an ADR in the case of a nonintervention. Our analyses included 144 interventions from the first general assessment and a selection of 90 consistently rated 'positive' interventions (from all assessments). RESULTS: On average, one in 200 prescriptions (0.49%) was found to have been positively modified by Dutch community pharmacists. About half of these interventions (49.8%) were aimed at preventing ADRs; 29.2% were rated as a positive modification in the effectiveness of pharmacotherapy and 8.6% affected both effectiveness and ADR. Reviewers' ratings varied widely between different categories of drug-related problems (DRPs). The impact of individual interventions (n = 83) varied, and for 53% of these interventions it was estimated to be relatively high. CONCLUSIONS: Pharmacists' interventions led to modification of prescriptions for an array of DRPs. Such interventions can contribute positively to the quality of pharmacotherapy. By extrapolating our data, we estimated a daily occurrence of approximately 2700 positive interventions in all Dutch pharmacies (1.6 per pharmacy per day). Reviewers rated the impact of interventions on a patient's health as significant in a substantial number of cases. PMID- 15521899 TI - Clinical and functional responses to salbutamol inhaled via different devices in asthmatic patients with induced bronchoconstriction. AB - AIMS: This study aimed at evaluating changes in airway patency, lung volumes and perception of breathing discomfort intensity following salbutamol administration via the Diskus dry-powder inhaler (DPI) or a pressurized metered-dose inhaler with the Volumatic valved holding chamber (pMDI + Volumatic) in asthmatic patients with methacholine-induced bronchoconstriction. METHODS: On six different study days, 18 patients inhaled methacholine until forced expiratory volume in 1 s (FEV(1)) decreased by approximately 35% of baseline. Following placebo, 200 and 400 microg of salbutamol through the pMDI + Volumatic or the Diskus, changes in FEV(1), volume-adjusted mean forced expiratory flow from 25 to 75% of the forced vital capacity (isoFEF(25-75)), lung volumes and breathing discomfort intensity, assessed by visual analogue scale (VAS) score, were repeatedly measured over a 60 min observation period. RESULTS: Induced bronchoconstriction was accompanied by obvious reductions in lung volumes and increases in VAS score. After salbutamol administration, FEV(1) and VAS score changes were similar in all experimental conditions. However, following 400 microg salbutamol via pMDI + Volumatic, isoFEF(25-75) values increased up to 4.48 l s(-1) (95% confidence interval 4.06, 4.90), a significantly (P < 0.01) higher value than those attained in all other experimental conditions. Independently of the salbutamol dose, lung volumes rose to significantly (P < 0.01) higher levels in pMDI + Volumatic than in Diskus trials. The low salbutamol dose via the pMDI + Volumatic and the high dose via the DPI increased isoFEF(25-75) and lung volumes to similar extents. CONCLUSIONS: Salbutamol via the pMDI + Volumatic provides greater isoFEF(25-75) and lung volume increases in asthmatic patients with induced bronchoconstriction; salbutamol-induced changes in VAS scores poorly reflect those in small airway patency. The lack of differences in FEV(1) increases observed after 200 and 400 microg salbutamol may reflect attainment of the flat portion of the dose-response curve using either device. PMID- 15521900 TI - Comparison of gentamicin dose estimates derived from manual calculations, the Australian 'Therapeutic Guidelines: Antibiotic' nomogram and the SeBA-GEN and DoseCalc software programs. AB - AIM: To compare gentamicin dose estimates from four predictive methods. METHODS: A retrospective study was conducted, comprising patients at Fremantle Hospital who received gentamicin therapy and had at least one gentamicin serum concentration reported. A manual calculation method, the Australian 'Therapeutic Guidelines: Antibiotic' (TGA) nomogram and the SeBA-GEN and DoseCalc software packages were compared. SeBA-GEN dose estimates were regarded as the reference standard. RESULTS: There were 64 males and 30 females with mean age of 58 +/- 16 years. In patients with moderate renal impairment (CL(Cr) = 30-60 ml min(-1); n = 21), mean dose estimates using DoseCalc and the manual calculation method were comparable to SeBA-GEN but the mean TGA nomogram dose (230 mg; 95% confidence interval 179, 281) was significantly lower than SeBA-GEN (286 mg; 261, 311; P = 0.002; one-way RM anova). In patients with mild renal impairment (CL(Cr) = 60-90 ml min(-1); n = 48), DoseCalc (392 mg; 367, 427) was comparable to SeBA-GEN (377 mg; 362, 392). Although the manual method (341 mg; 306, 376; P = 0.007) and the TGA nomogram (335 mg; 302, 368; P < 0.001) estimates were significantly lower than SeBA-GEN, the practical difference was modest. CONCLUSIONS: SeBA-GEN and DoseCalc are generally comparable for estimation of gentamicin doses in patients with renal impairment. The 'Therapeutic Guidelines: Antibiotic' nomogram is a valid approach to dosage estimation, but only when used in patients with normal renal function. Simple manual calculations are a suitable alternative in patients with renal impairment. PMID- 15521901 TI - Association between adherence to statin therapy and lipid control in Hong Kong Chinese patients at high risk of coronary heart disease. AB - AIMS: To examine the pattern of adherence to statin therapy and to determine the association of adherence to statin therapy and the control of serum low-density lipoprotein (LDL)-cholesterol in a cohort of Hong Kong Chinese patients at high risk of coronary heart disease (CHD). METHODS: This was a prospective observational cohort study conducted at the outpatient clinics of a public teaching hospital in Hong Kong. Patients at high risk of CHD who had been initiated on statin monotherapy for < 12 months were recruited. The statin prescription was dispensed in a bottle with the Medication Event Monitoring System (MEMS). Adherence was assessed in two dimensions: dose-count was defined as the percentage of doses taken, and dose-time was defined as the percentage of doses taken within the suggested time interval. Lipid profiles were obtained at baseline and during two follow-up visits at month 3 and month 6. RESULTS: Eighty three patients completed the study. The median adherence to dose-count and to dose-time were 95% (25-75th percentile = 87-99%) and 78% (25-75th percentile = 17 92%), respectively. Both dose-count and dose-time adherence declined slightly over the first 6 months of therapy. Living with family [relative risk (RR) = 0.79, 95% confidence interval (CI) 0.63, 0.91] and duration of therapy (RR = 0.99, 95% CI 0.98, 1.00) were negative predictors while number of family members (among those living with family) (RR = 1.05, 95% CI 1.00, 1.08) was a positive predictor for adherence to dose-count. Monthly household income (RR = 1.01, 95% CI 1.00, 1.02) and angina (RR = 1.29, 95% CI 1.05, 1.58) were positive predictors while living with family (RR = 0.74, 95% CI 0.55, 0.90) was a negative predictor for dose-time adherence. Percent reduction in serum LDL-cholesterol was correlated to dose-count (P < 0.001) and dose-time (P = 0.047) adherence. Statistically significant correlations were observed between adherence to dose count and LDL reduction (R(2) = 0.130; P = 0.001), and between dose-time adherence and LDL reduction (R(2) = 0.048; P = 0.047). CONCLUSION: High adherence to statin therapy was found in a cohort of Chinese patients at high risk of CHD and the adherence declined slightly over time. A weak association between adherence to statin dose-count and LDL reduction and a marginal association between dose-time adherence and LDL reduction were observed. PMID- 15521902 TI - Comparison of tincture of opium and methadone to control opioid withdrawal in a Thai treatment centre. AB - AIMS: To evaluate the effectiveness of oral tincture of opium (TOP) and methadone to control opioid withdrawal in patients in northern Thailand. METHODS: Open label, parallel group study in an inpatient facility compared 15 former heroin users receiving methadone 5-20 mg 12 hourly with 15 former opium smokers receiving TOP (3.33-10 mg morphine equivalents 12 hourly). At 0, 1, 3 and 8 h, blood, withdrawal scores and subjective opioid effects were collected. RESULTS: There was a reciprocal association between withdrawal scores/direct subjective opioid effects and plasma (R)-methadone, but not plasma morphine, concentrations. Withdrawal scores at the time of dosing were higher in the TOP patients (9.1 +/- 3) than in the methadone patients (4.5 +/- 4.6) and in the TOP patients were significantly (P = 0.001) attenuated at 3 and 8 h. CONCLUSIONS: At the doses used, TOP was inferior to methadone in suppressing withdrawal. It could prove to be a cost effective and valuable drug, but only after dose size and frequency are further investigated. PMID- 15521903 TI - A randomized, double-blind, placebo controlled trial of melatonin add-on therapy in epileptic children on valproate monotherapy: effect on glutathione peroxidase and glutathione reductase enzymes. AB - AIMS: To compare the effect of add-on melatonin with placebo on the antioxidant enzymes (glutathione peroxidase and glutathione reductase) in epileptic children on valproate monotherapy. METHODS: In a double-blind, randomized, placebo controlled trial, the effect of add-on melatonin administration on the antioxidant enzymes in epileptic children on valproate (VPA) monotherapy was assessed. A total of 31 patients met the entry criteria. 16 patients were randomly allocated to receive add-on melatonin, and 15 to receive add-on placebo. Blood samples (5 ml) were collected just before the morning dose of valproate for baseline values of glutathione peroxidase and glutathione reductase enzymes, and then after 14 days of add-on melatonin/placebo. Blood was then centrifuged at 3500 r.p.m., serum separated and stored in deep freezer at -20 degrees C until assay of glutathione reductase. Heparinized blood was collected and stored at -20 degrees C in eppendorfs in the deep freezer for assay of glutathione peroxidase. All activity assays were performed on the Ames (Technicon) RA 50 chemistry analyser. RESULTS: Fifteen patients in the add-on melatonin group and 14 patients in the add-on placebo group were finally assessed. There was an increase in the activity of antioxidant enzymes, glutathione peroxidase (GSH-Px) and glutathione reductase (GSSG-Rd), in the add-on melatonin (MEL) group as compared with a reduction in the same in the add-on placebo group (P). After the addition of melatonin/placebo in the respective groups, there was a 7.5% decrease in GSH-Px in the valproate + placebo group, whereas a 11.9% increase in the valproate + melatonin group was observed, the difference between the groups being not statistically significant (P = 0.29). On administration of melatonin/placebo, the post-treatment concentrations of GSSG-Rd in the valproate + placebo group decreased from 92.0 U l(-1) to 67.0 U l(-1) and increased from 82.0 U l(-1) to 113.0 U l(-1), in the valproate + melatonin group, respectively, the difference between them being statistically significant (P = 0.05). The percentage change in the values of GSSG-Rd in the two groups was statistically significant (P = 0.005). CONCLUSIONS: Melatonin exerts neuroprotection due to its antioxidant, antiexcitotoxic and free radical scavenging properties within the central nervous system. Melatonin, thus, as an adjunct, can be a putative neuroprotector in conditions involving oxidative stress like epilepsies. PMID- 15521904 TI - MDR1 haplotypes derived from exons 21 and 26 do not affect the steady-state pharmacokinetics of tacrolimus in renal transplant patients. AB - AIM: This retrospective study investigated the influence of MDR1 haplotypes derived from the polymorphisms 2677G > T (exon 21) and 3435C > T (exon 26) on the pharmacokinetics of the immunosuppressant drug tacrolimus in 73 renal transplant patients. METHODS: Based on both variants of SNPs 2677 and 3435, four different haplotypes and eight different genotypes were identified in the study sample. Tacrolimus trough concentrations (C(0)) were compared between different SNP variants and genotypes, as well as between carriers and noncarriers of each haplotype. Additionally, CYP3A5 genotype (6956G > A) was determined. RESULTS: No significant differences were observed between groups. Differences in mean tacrolimus C(0) values between carriers and noncarriers of each haplotype ranged from -0.04 microg/litre (95% confidence interval: -0.53 to 0.60) to -23 microg/litre (-1.07 to 1.53). No association was found between CYP3A5*1/*3 genotype and tacrolimus Co concentractions. CONCLUSION: MDR1 haplotypes derived from the SNPs 2677G > T (exon 21) and 3435C > T (exon 26) do not influence the pharmacokinetics of tacrolimus in renal transplant patients. PMID- 15521905 TI - Tolerability of spironolactone in patients with chronic heart failure -- a cautionary message. AB - AIM: To assess how well heart failure patients tolerate spironolactone in routine clinical practice. DESIGN: Retrospective analysis of 226 patients attending a specialist heart failure clinic. RESULTS: One hundred and thirty of 226 (57.5%) patients tried spironolactone at least once. Forty-four of 130 (33.8%) discontinued spironolactone due to side-effects after a mean of 11.1 months; 59/141 (41.8%) trials of spironolactone resulted in at least one side-effect; therapy was stopped in 30/141 (21.3%) trials due to raised potassium or creatinine. Significant risk factors for raised potassium/creatinine were age and baseline potassium level. CONCLUSIONS: Potentially serious side-effects are common despite appropriate use of spironolactone. PMID- 15521906 TI - 'Ally McBeal heart?'-- drug induced cardiomyopathy in a young woman. PMID- 15521907 TI - Drug-induced pancreatitis: lessons in data mining. PMID- 15521911 TI - Pancytopenia in hereditary haemorrhagic telangiectasia. PMID- 15521912 TI - Acute lymphoblastic leukaemia relapsing with solid hepatic tumours after bone marrow transplantation. PMID- 15521913 TI - Point of care testing for INR monitoring: where are we now? AB - Point of care (POC, or near patient) testing for measurement of the international normalized ratio (INR) has facilitated the devolution of service delivery from the traditional hospital outpatient setting. However it must be undertaken within the confines of safe practice involving quality control procedures. The evaluation of INR POC tests should be closely related to the clinical issues of management and, specifically, improving the quality of care. One benefit of POC testing is in the increased motivation that some practitioners feel, being able to perform diagnostic tests without sending samples to a laboratory. POC for INR testing within primary care eliminates the delay in waiting for the result to be processed by the hospital laboratory, and the subsequent delay in informing the patient of their dosing advice. This review describes the utilization of POC testing outside the laboratory setting to develop models of care for oral anticoagulation management. PMID- 15521914 TI - Inhibitors in congenital coagulation disorders. AB - The development of inhibitory 'allo' antibodies to a deficient coagulation factor is arguably now the most severe and important complication of clotting factor concentrate exposure in haemophilia and other congenital coagulation disorders. Furthermore, development of an inhibitor to the factor VIII or factor IX transgene product remains a significant concern in gene therapy protocols for haemophilia. Although the development of an inhibitor does not usually change the rate, initial severity or pattern of bleeding, it does compromise the ability to manage haemorrhage in affected individuals, resulting in a greater rate of complications, cost and disability. The purpose of this review is to summarize current understanding of the epidemiology, immunobiology, laboratory evaluation and management of inhibitors arising in patients with congenital coagulation disorders. An attempt has been made to focus on recent advances in the immunology of inhibitors, and to speculate on their potential clinical application. PMID- 15521915 TI - Breast cancer resistance protein (BCRP/MXR/ABCG2) in adult acute lymphoblastic leukaemia: frequent expression and possible correlation with shorter disease-free survival. AB - Drugs used in treatment of adult acute lymphoblastic leukaemia (ALL) are substrates for breast cancer resistance protein (BCRP, MXR, ABCG2), which may thus play a role in resistance in this disease. Pretreatment blasts from 30 adult ALL patients were studied for BCRP mRNA by quantitative reverse transcription polymerase chain reaction analysis, BCRP protein by immunophenotyping with three antibodies and BCRP function by fumitremorgin C modulation of intracellular mitoxantrone retention, measured by flow cytometry. BCRP mRNA in all cases encoded wild type protein (BCRP(R482)), which mediates mitoxantrone and methotrexate resistance, but only low-level anthracycline resistance. The BXP-21, BXP-34 and anti-ABCG2 antibodies stained blasts in 13, 11 and 14 cases (43%, 37% and 47%); BXP-21 correlated well with BXP-34 and anti-ABCG2, but BXP-34 and anti ABCG2 did not correlate, and antibody staining did not correlate with mRNA levels. BCRP function was seen in 21 cases (70%), but correlated poorly with antibody staining. An exploratory statistical analysis indicated that BXP-21 staining was predictive of shorter disease-free survival (DFS) (P = 0.0374) in this small patient population. Poor correlations between mRNA, protein and function indicate the complex biology of BCRP in adult ALL, and the possible correlation of BCRP expression with DFS should be studied in larger series. PMID- 15521916 TI - Erythropoietin treatment of the anaemia of myelofibrosis with myeloid metaplasia: results in 20 patients and review of the literature. AB - Recombinant human erythropoietin (rHuEPO) is an effective treatment for the anaemia that occurs secondary to various conditions, but its role in myelofibrosis with myeloid metaplasia (MMM) is not well established. rHuEPO, at an initial dose of 10 000 U thrice a week, was given to 20 patients with MMM and anaemia. Complete response (CR) was defined as transfusion cessation with normal haemoglobin (Hb) levels and partial response (PR) as a transfusion decrease > or =50% and Hb > 10 g/dl maintained for at least 8 weeks. Nine patients (45%) showed a favourable response to treatment, including four CR and five PR, four of whom have maintained their response at a median follow-up of 12.5 months (range: 4-21 months) from the start of treatment. The pretreatment factors associated with a favourable response were lack of transfusion requirement (P = 0.002) and higher Hb at start treatment of (P = 0.01). An analysis of the present series (n = 20) and 31 patients from the literature identified 28 (55%) favourable responses to rHuEPO, including 16 CR and 12 PR. In the multivariate analysis, serum erythropoietin level <125 U/l was found to be associated with a favourable response to rHuEPO, whereas lack of transfusional support had borderline significance. PMID- 15521917 TI - CD40 stimulation of B-cell chronic lymphocytic leukaemia cells enhances the anti apoptotic profile, but also Bid expression and cells remain susceptible to autologous cytotoxic T-lymphocyte attack. AB - To enhance the poor antigen-presenting capacity of B-cell chronic lymphocytic leukaemia (B-CLL), CD40 triggering has been considered as an active immunotherapy. However, CD40 stimulation also has an anti-apoptotic effect and may further impair the dysregulated response of B-CLL to apoptotic stimuli. Therefore, we measured the expression of virtually all regulators of apoptosis before and after CD40 stimulation. These findings were correlated with sensitivity for chemotherapy- and death-receptor-induced apoptosis and T-cell mediated killing. CD40 stimulation enhanced the constitutive anti-apoptotic profile of B-CLL cells by upregulation of Bcl-xL and Bfl-1 and downregulation of the BH3-only protein Harakiri. Unexpectedly, the BH3-only protein Bid was strongly induced. Functionally, CD40-stimulated B-CLL cells became resistant to drug-induced apoptosis and, despite upregulation of CD95 and Bid, were not sensitive to CD95L. In contrast, autologous T cell killing, triggered by loading CLL cells with viral (CMV) peptides, was very efficient both before and after CD40 stimulation. Upon CTL interaction, CLL targets underwent mitochondrial depolarization and caspase-3 activation. Thus, despite an increased anti apoptotic profile, CD40 triggered B-CLL cells remain excellent targets for resident cytotoxic T cells. These data support therapeutic exploitation of CD40 stimulation in B-CLL, provided that a strong CTL component is induced. PMID- 15521918 TI - Role of CD21 antigen in diffuse large B-cell lymphoma and its clinical significance. AB - Recent advances in immunological and molecular technology have prompted proposals to change tumour classification and treatment strategies. Cell surface antigens are now easy to access, and tumour origins and clinical characteristics are now readily identifiable. However, in diffuse large B-cell lymphoma (DLBCL), one of the heterogeneous forms of haematological malignancy, the clinical significance of tumour surface antigens has not been well documented. We analysed the tumour surface antigens of 50 tumours from newly diagnosed DLBCL patients by flow cytometry in accordance with their clinical characteristics and followed the patients for a median 3.7 years. Statistical analysis showed that CD21 expression was significantly negatively associated with mortality in DLBCL (CD21 negative versus positive; relative risk = 2.36, P < 0.05). As a result of these clinical observations, we generated CD21-overexpressed (CD21(+)) lymphoma cell lines after gene transfection and analysed tumour cell growth in vivo in immunocompromised mice. Mice challenged with vector-only transfectants and parental cells as controls died within 50 d. In contrast, mice injected with CD21(+) transfectants exhibited significantly reduced tumour growth and 83% survived long term (versus control groups; P < 0.05). Interestingly, all established CD21(+) transfectants (six clones from different bulks) showed homotypic aggregation during in vitro cell culture, and anti-CD21 antibodies did not block this aggregation. Expression of CD21 is strongly associated with increased survival in DLBCL in vivo. CD21 expression may be indirectly concerned with the expression of additional cell adhesion molecules. PMID- 15521919 TI - Detection of p53 dysfunction by flow cytometry in chronic lymphocytic leukaemia. AB - In chronic lymphocytic leukaemia (CLL) cells, functional impairment of the p53 pathway is detectable by Western blotting as impaired up-regulation of p21 (a transcriptional target of p53) in response to ionizing radiation (IR). The type A defect is characterized by baseline p53 overexpression and is associated with TP53 mutation. The type B defect is characterized by impaired IR-induced p53 up regulation and is associated with inactivation of the ataxia telangiectasia mutated gene (ATM). Both abnormalities are strongly associated with adverse clinical outcome. In the present study, flow cytometry was found to be an effective alternative to Western blotting in the detection of p53 dysfunction in CLL. PMID- 15521920 TI - T-cell post-transplantation lymphoproliferative disorders after cardiac transplantation: a single institutional experience. AB - Post-transplantation lymphoproliferative disorders (PTLDs) are a well-recognized and potentially life-threatening complication of solid organ transplantation. While the vast majority of PTLDs are B-cell lymphoproliferations, T-cell PTLDs are rarely seen. Among 898 patients receiving cardiac transplants between 1990 and 2003, 34 patients (3.8%) developed PTLDs with two (0.2%) T-cell PTLDs, 31 (3.5%) B-cell PTLDs and one (0.1%) natural killer cell PTLD. An additional three cases of T-cell PTLD were identified among all cardiac transplant patients followed at our institution. These T-cell PTLDs comprised a heterogeneous group of Epstein-Barr virus negative lymphoproliferations that developed late after transplantation and followed an aggressive course. PMID- 15521921 TI - von Willebrand factor cleaving protease (ADAMTS-13) and ADAMTS-13 neutralizing autoantibodies in 100 patients with thrombotic thrombocytopenic purpura. AB - The congenital or acquired deficiency of the von Willebrand factor (VWF) cleaving protease, ADAMTS-13 has been specifically associated with a diagnosis of thrombotic thrombocytopenic purpura (TTP), a microangiopathy characterized by the formation of occlusive platelet thrombi. The mechanisms of TTP were investigated in 100 patients diagnosed on the basis of the presence of at least three of the following: thrombocytopenia, haemolytic anaemia, elevated serum levels of lactate dehydrogenase and neurological symptoms. Plasma levels of ADAMTS-13 were severely reduced (<10% of normal) in 48%, moderately reduced (between 10% and 46%) in 24% and normal (>46%) in 28%. A neutralizing antibody was the cause of the deficiency in 38% of the cases, with a higher prevalence of this mechanism (87%) in the 48 patients with severely reduced ADAMTS-13. Double heterozygosity for a 29 base pair (bp) deletion and a nucleotide insertion and homozygosity for a 6 bp deletion in the ADAMTS13 gene were identified only in two patients born from consanguineous marriages. In conclusion, this study indicated that ADAMTS-13 was normal in nearly one-third of patients with TTP and that ADAMTS-13 deficiency was not associated with the presence of neutralizing antibodies in more than half of the patients. PMID- 15521922 TI - TAFI polymorphisms at amino acids 147 and 325 are not risk factors for cerebral infarction. AB - Thrombin-activatable fibrinolysis inhibitor (TAFI) was reported as an anaphylatoxin-inactivating enzyme generated by proteolytic cleavage of its zymogen, and is the same enzyme as that first designated by our group as procarboxypeptidase R (proCPR). Its level in plasma appears to influence vascular disease. In addition, TAFI activity is strongly influenced by genetic polymorphism, especially at amino acids 147 and 325. We investigated whether these TAFI polymorphisms would act as a risk factor for cerebral infarction (CI) by examining 253 samples in which the diagnosis was cliniconeuropathologically confirmed. We found little that was statistically significant in terms of these polymorphisms among patients with no vascular problems or in a population-based control group. In the present study of an elderly Japanese group, our samples revealed a lower percentage of the Ile allele at Thr/Ile-325 compared with western counterparts. Although patients with severe infarcts had a lower percentage of the Ile allele (10%) at amino acid position 325 compared with the slightly and moderately affected patients and the population-based control group (15-18%), no statistical significance was found. None of our results showed any statistical correlation between TAFI polymorphisms and CI. PMID- 15521923 TI - Low-intensity warfarin reduces thrombin generation and fibrin turnover, but not low-grade inflammation, in men at risk of myocardial infarction. AB - In the Thrombosis Prevention Trial (TPT), low-intensity warfarin reduced the risk of first coronary events only when the achieved international normalized ratio (INR) was > or =1.4. To validate the likely mechanism of action of low-intensity warfarin we measured its effects on plasma markers of thrombin generation, fibrin turnover and low-grade inflammation in TPT participants. D-dimer and prothrombin fragment F1.2 levels were lower at INRs > or =1.4 (P = 0.02 and 0.03 respectively); levels fell as INR increased (P for trend 0.04 and 0.002 respectively). C-reactive protein did not vary with INR. The efficacy of warfarin is related to reductions in thrombin generation and fibrin turnover. PMID- 15521924 TI - Clonogenicity, gene expression and phenotype during neutrophil versus erythroid differentiation of cytokine-stimulated CD34+ human marrow cells in vitro. AB - With the objective to correlate clonogenicity, gene expression and phenotype during differentiation, human bone marrow CD34(+) cells were cultured in vitro to stimulate erythroid or neutrophil development, and sorted into five subpopulations according to their surface expression of CD15/CD33 and blood group antigen A/CD117 respectively. Sorted cells were cultured in methylcellulose and analysed by real-time reverse transcription polymerase chain reaction for expression of neutrophil and erythroid marker genes. Surface expression of CD15 coincided with restriction to neutrophil/monocyte differentiation and A antigen with restriction to erythroid differentiation. GATA-2 mRNA was down-regulated during both neutrophil and erythroid maturation, whereas GATA-1, SCL, ABO, erythropoietin receptor, Kell, glycophorin A, beta-globin and alpha-haemoglobin stabilizing protein were up-regulated during erythroid differentiation and silenced during neutrophil differentiation. CCAAT/enhancer-binding protein (C/EBP)-alpha, PU.1, granulocyte colony-stimulating factor receptor, PR3, C/EBP epsilon and lactoferrin were sequentially expressed during neutrophil differentiation but rapidly down-regulated during the early erythroid stages. Nuclear factor erythroid-derived 2 (NF-E2) and glycophorin C were expressed both during neutrophil and erythroid differentiation. Our data support the notion of early expression of several lineage-associated genes prior to actual lineage commitment, defined by surface expression of CD15 and A antigen as markers for definitive neutrophil/monocyte and erythroid differentiation respectively. Previous findings, primarily from cell lines and mouse models, have been extended to adult human haematopoiesis. PMID- 15521925 TI - Analyses for binding of the transferrin family of proteins to the transferrin receptor 2. AB - Transferrin receptor 2 alpha (TfR2 alpha), the major product of the TfR2 gene, is the second receptor for transferrin (Tf), which can mediate cellular iron uptake in vitro. Homozygous mutations of TfR2 cause haemochromatosis, suggesting that TfR2 alpha may not be a simple iron transporter, but a regulator of iron by identifying iron-Tf. In this study, we analysed the ligand specificity of TfR2 alpha using human transferrin receptor 1 (TfR1) and TfR2 alpha-stably transfected and expressing cells and flow-cytometric techniques. We showed that human TfR2 alpha interacted with both human and bovine Tf, whereas human TfR1 interacted only with human Tf. Neither human TfR1 nor TfR2 alpha interacted with either lactoferrin or melanotransferrin. In addition, by creating point mutations in human TfR2 alpha, the RGD sequence in the extracellular domain of TfR2 alpha was shown to be crucial for Tf-binding. Furthermore, we demonstrated that mutated TfR2 alpha (Y250X), which has been reported in patients with hereditary haemochromatosis, also lost its ability to interact with both human and bovine Tf. Although human TfR1 and TfR2 alpha share an essential structure (RGD) for ligand-binding, they have clearly different ligand specificities, which may be related to the differences in their roles in iron metabolism. PMID- 15521926 TI - Non-T-cell-depleted HLA-haploidentical stem cell transplantation after reduced intensity conditioning in advanced haematological malignancies based on feto maternal microchimerism. PMID- 15521927 TI - Clinical audit and clinical governance. PMID- 15521928 TI - Developments in treatment of primary irresectable rectal cancer. AB - Abstract The treatment options for primary irresectable rectal cancers are discussed. Assessment of tumour stage is the first step for an appropriate choice of treatment. Following a diagnosis of rectal cancer, a vast array of diagnostic procedures is available to determine its stage, and thereby its best treatment options. From the many (new) diagnostic options the merits and drawbacks are discussed. If a diagnosis of irresectability is made, further treatment options should include radiotherapy in most cases, some aspects of timing and application, i.e. intra-operative treatment are discussed. Chemotherapy options are manifold, the results are discussed and some new options are explored. PMID- 15521929 TI - Impact of national guidelines about the management of colorectal cancer on Australian surgeons' awareness of evidence: a pre/post survey. AB - OBJECTIVE: To evaluate the immediate impact of national evidence-based guidelines about colorectal cancer on Australian surgeons' self-reported practice and their deficits in awareness of scientific evidence underpinning clinical management practices. DESIGN: Pre/post evaluation, comprising preguidelines survey (November 1998) and postguidelines survey (February 2001). METHODS: One hundred and fourteen Australian surgeons returned postguidelines surveys, of whom 103 (90%) agreed to matching of their pre- and postguidelines responses. National distribution of the CRC guidelines occurred in November 1999. Over the ensuing year, dissemination strategies included seminars, presentations at conferences and journal articles. The main outcome measures used were changes in awareness of evidence for each of 23 clinical recommendations, changes in overall awareness score (maximum possible 23), changes in subscore for nine items for which evidence was compelling and predictors of change. RESULTS: Of those surgeons followed up, 95% were aware of the guidelines and 32% had read them in detail. Only 47% recalled the consumer version. The three most highly rated topics in the guidelines were: high-risk familial syndromes (45%); screening based on family history of colorectal cancer (40%); population screening for colorectal cancer (25%). Compared with baseline, there was a modest improvement in the mean overall awareness score (P = 0.02). Paired analyses of awareness of the evidence for each of 23 individual topics revealed significant improvement only in five. For two, awareness significantly decreased. CONCLUSIONS: Our pre/post findings are not inconsistent with the expectation that dissemination of the CRC guidelines has had some short-term impact. However, definitive evidence acquired through more rigorously designed controlled trials will be needed to determine first, whether surgical practice has changed and, second, whether implementation of the CRC guidelines or some other secular event caused such change. PMID- 15521930 TI - A study on the routes of referral for patients with colorectal cancer and its affect on the time to surgery and pathological stage. AB - OBJECTIVE: Firstly, to determine the proportion of colorectal cancer (CRC) patients seen within an established two week rule (TWR) system and to observe other routes of referral for CRC patients. Secondly to determine if referral route affects the interval to, and cancer stage at, definitive treatment. PATIENTS AND METHODS: GP referrals of patients with CRC were divided into direct surgical outpatient referrals (group 1) and indirect referrals to accident and emergency and medical outpatients (group 2). Data were recorded on the time to definitive treatment and the location and stage of tumour. RESULTS: There were 78 patients in group 1. Thirty (20%) patients were referred by their GP under the TWR, 31 (21%) as urgent and 17 (12%) as nonurgent referrals, to surgical outpatients. There were 69 patients in group 2. Forty-two (29%) were referred initially to the accident and emergency department and 27 (18%) to general medical outpatients. Group 1 patients were treated within a median of 70.5 days and group 2 patients within 14 days of referral (P < 0.0005). Group 2 contained tumours of a significantly more advanced pathological stage (P = 0.015) and more proximal colonic cancers (P < 0.005). CONCLUSION: Fifty-three percent of patients with CRC were referred directly to surgical outpatients, 20% under the TWR guidelines. Despite having this system in place direct referrals were slower to treatment but the tumours were still of a less advanced pathological stage. Compliance with the TWR should not be used as a means of assessing a colorectal unit's treatment of CRC. PMID- 15521931 TI - Residual urine volume after total mesorectal excision: an indicator of pelvic autonomic nerve preservation? Results of a case-control study. AB - OBJECTIVE: The rate of bladder dysfunctions after total mesorectal excision (TME) for rectal cancer can be decreased by bilateral pelvic autonomic nerve preservation (PANP). However, it is not clear yet, how often partial nerve impairment may lead to bladder dysfunction. It was the aim of a case-control study, to examine the residual urine volume in patients before and after TME with and without complete PANP, in order to clarify, whether this parameter allows conclusions on the quality of PANP. PATIENTS AND METHODS: Regarding bladder function, a case group (n = 26) without complete PANP was compared with a control group (n = 26) with complete identification and nerve preservation according to standadized intra-operative documentation. Twenty-six match pairs were established, identical regarding gender, wall infiltration depth, tumour site, operation procedure and operation extent. Rates of neoadjuvant therapy, R0 classification, anastomotic leakage, wound and urinary tract infection were equally distributed for both the case- and control group (P > 0.05). Residual urine volume was pre- and post-operatively determined by sonography. RESULTS: Pre operatively, residual urine volumes differed neither between the pairs nor between both groups with and without nerve preservation. In the case group with incomplete PANP there was a difference between pre- and post-operative residual urine volume (median; quartil: 2.5 ml; 0.0-32.5 ml vs 130 ml; 0.0-317 ml; P = 0.001). In the control group there was no difference (median; quartile: 0.0 ml; 0.0-20 ml vs 15.5 ml; 0.0-62.0 ml; P = 0.07). The difference between the postoperatively measured volumes of the case and control group were significant (P = 0.001). With residual urine volume = 100 ml, the risk of incomplete PANP was 14 times higher (odds ratio). CONCLUSIONS: Residual urine volume is an indicator of the completeness of PANP during TME. It should be determined pre- and post operatively, and besides the recording of the neurogenic bladder, serve as a quality control. PMID- 15521932 TI - Long strictureplasty is as safe and effective as short strictureplasty in small bowel Crohn's disease. AB - BACKGROUND: For the past 20 years it has been shown that intestinal strictureplasty is safe and effective in the management of short strictureplasty. Long strictureplasty (> 20 cm) may be an alternative to resection in some patients, especially in diffuse disease or after previous extensive resections. We reviewed the outcome of long strictureplasty for Crohn's Disease, to examine safety and recurrence rates, compared with conventional short strictureplasty. METHODS: Sixty-two patients have undergone single strictureplasty for jejunoileal Crohn's disease since 1974. Median follow-up was 121 (range 7-253) months. Twenty one operations involved a single long strictureplasty, and 41 operations had a single short strictureplasty. RESULTS: No significant differences were identified between the groups. The postoperative complications in long strictureplasty included 2 abscesses only and after short strictureplasty there was one anastomotic leak and one postoperative abscess. The median hospital stay was 10 days after long strictureplasty and 9 days after short strictureplasty. Three-, 5 and 10-year disease-free rates for long and short strictureplasty, respectively, were 3-year 80.4% and 62.1%; 5-year 55.2% and 49.8% and 10-year 49.1% and 33.5% (NS). CONCLUSIONS: These data indicate that long strictureplasty is safe and produces equivalent results to conventional (short) strictureplasty. PMID- 15521933 TI - The strength duration test: a novel tool in the identification of occult neuropathy in women with pelvic floor dysfunction. AB - OBJECTIVE: The Strength-duration test (SDT) is a simple minimally invasive measure of muscle innervation, recently adapted for the assessment of the external anal sphincter (EAS). This test can discriminate women with faecal incontinence from controls. The purpose of this study was to determine if the SDT could detect denervation of the EAS in women with weak but anatomically intact EAS and normal pudendal nerve terminal motor latency (PNTML). PATIENTS AND METHODS: Thirteen women with weak but intact EAS on endoanal ultrasound (EAUS) with normal maximum resting pressure and PNTML were recruited to undergo the SDT. The results from this group were compared to control data for the SDT previously collected in our unit. RESULTS: Nine of 13 women were found to have SDT above a 95% confidence interval of the mean curve for controls and six had a SDT outside a 90% confidence interval for individuals derived from control data, suggesting denervation of the EAS. The mean area under the curve was significantly higher in our study group compared to controls (91.0 microsmA vs 72.2 microsmA, P = 0.047) as was the current intensity measured at the 1 ms pulse duration (18.2 mA vs 12.94 mA, P = 0.048), typical of denervation with this test. CONCLUSION: The SDT was abnormal in nine of the 13 study patients. This may partly explain reduced maximum voluntary contraction seen in this group of patients. SDT may be a more sensitive tool in detecting neuropathy than latency measurement. PMID- 15521934 TI - Local resection of rectal tumours using the Salvati operating proctoscope--a safe and effective technique. AB - BACKGROUND: Per-anal excision can be an effective method for treating large rectal adenomas and selected rectal cancers. This procedure is suitable for adenomas that are too large for colonoscopic excision and for early rectal cancers in patients that are unfit for major resection. PATIENTS AND METHODS: We retrospectively reviewed 37 patients (16 male, 21 female) that had a local resection using a Salvati operating proctoscope. Fifteen had rectal cancer and 22 had rectal adenoma and all have been followed-up for a median of 14 months (range 2-65). Most cancers were staged pre-operatively with endorectal ultrasound and 6 cancer patients received adjuvant therapy. RESULTS: There were no intra-operative complications, 6 minor postoperative complications, none requiring re intervention (1 peri-anal haematoma, 2 postoperative anaemia, 2 chest complications, 1 secondary haemorrhage) and no peri-operative deaths. Lesions were histologically completely excised in 33 (89%). Thirty-four (92%) did not have any recurrence, 2 (5%) adenomas recurred (both successfully treated with further local resection) and 1 (3%) failed to attend for follow-up. Six have since died, all from comorbid conditions. CONCLUSION: Local resection of rectal tumours using the operating proctoscope is a low cost technique with a low complication rate. Outcome of this treatment is good, and similar to other surgical modalities. Local tumour recurrences are uncommon and can be successfully treated with further local treatment. PMID- 15521935 TI - Blood loss and transfusion after total mesorectal excision and conventional rectal cancer surgery. AB - OBJECTIVES: A recent study showed less bleeding and need of transfusion after total mesorectal excision (TME) compared with conventional rectal cancer surgery. The aim of this study was to evaluate this result in more details. PATIENTS AND METHODS: Comparison of transfusion history in rectal cancer resections in two different multicentre-studies. Two hundred and forty-six patients were operated in the period 1991-93 with a conventional technique and 311 patients were operated with TME-technique in the period 1996-98. Peri-operative data, including blood transfusion from one month before until one month after the operation, was recorded prospectively. RESULTS: The median intra-operative blood loss was 1000 ml, range 50-6000 ml, before, and 550 ml, range 10-6000 ml (P < 0.001) after introduction of TME. The overall peri-operative transfusion rate was reduced from 73% to 43% (P < 0.001). When adjusted for blood loss, age, gender, weight, and type of resection, TME significantly reduced the risk of receiving intra or postoperative blood transfusion by 0.4 (CI: 0.3-0.6). The variability in blood loss among 12 TME-centres was more than 400% and not correlated with transfusion requirements within the centres. CONCLUSION: TME results in a reduced blood loss and a reduction of blood transfusion, but additional factors others than blood loss seems to influence the decision of transfusion. PMID- 15521936 TI - Laparoscopic proctocolectomy with restorative ileal-anal pouch. AB - OBJECTIVE: The aim of the study was to analyse the outcome of restorative proctocolectomy carried out by laparoscopic surgery. METHODS: A prospectively collected electronic database of all colorectal laparoscopic procedures performed between April 2001 and July 2003 has been used to identify surgical outcomes in 14 consecutive patients who have undergone laparoscopic RPC. RESULTS: Fourteen patients (5 male), median BMI 24 kg/m(2) have undergone restorative laparoscopic proctocolectomy over a two year period: 13 (ulcerative colitis, one with cancer) and 1 (FAP). The median operation time was 260 min; time has not decreased with experience. There were no intra-operative surgical complications or deaths. Patient controlled analgesia continued for a median of 36 h. The median time to diet was 48 h and median hospital stay 7 days; three patients required nasogastric aspiration for delayed gastric emptying. Eighteen regional lymph nodes were retrieved local to the carcinoma. There was one anastomotic leak. All covering stomas were closed by 6 months (12 by eight weeks). All 14 patients are fully continent, able to suppress urgency and have a median pouch frequency of 4/24 h. None admit to having problems with potency, orgasm sensation, ejaculation, micturition. One lady reports dysparunia. All are highly satisfied with functional outcome and cosmesis. CONCLUSION: We are encouraged to continue to offer our patients the option of a laparoscopic resection. PMID- 15521937 TI - Risk factors for anastomotic leakage after anterior resection of the rectum. AB - OBJECTIVE: Surgical technique and peri-operative management of rectal carcinoma have developed substantially in the last decades. Despite this, morbidity and mortality after anterior resection of the rectum are still important problems. The aim of this study was to identify risk factors for anastomotic leakage in anterior resection and to assess the role of a temporary stoma and the need for urgent re-operations in relation to anastomotic leakage. PATIENTS AND METHODS: In a nine-year period, from 1987 to 1995, a total of 6833 patients underwent elective anterior resection of the rectum in Sweden. A random sample of 432 of these patients was analysed (sample size 6.3%). The associations between death and 10 patient- and surgery-related variables were studied by univariate and multivariate analysis. Data were obtained by review of the hospital files from all patients. RESULTS: The incidence of symptomatic clinically evident anastomotic leakage was 12% (53/432). The 30-day mortality was 2.1% (140/6833). The rate of mortality associated with leakage was 7.5%. A temporary stoma was initially fashioned in 17% (72/432) of the patients, and 15% (11/72) with a temporary stoma had a clinical leakage, compared with 12% (42/360) without a temporary stoma, not significant. Multivariate analysis showed that low anastomosis (< or = 6 cm), pre-operative radiation, presence of intra-operative adverse events and male gender were independent risk factors for leakage. The risk for permanent stoma after leakage was 25%. Females with stoma leaked in 3% compared to men with stoma who leaked in 29%. The median hospital stay for patients without leakage was 10 days (range 5-61 days) and for patients with leakage 22 days (3-110 days). CONCLUSION: In this population based study, 12% of the patients had symptomatic anastomotic leakage after anterior resection of the rectum. Postoperative 30-day mortality was 2.1%. Low anastomosis, pre-operative radiation, presence of intra-operative adverse events and male gender were independent risk factors for symptomatic anastomotic leakage in the multivariate analysis. There was no difference in the use of temporary stoma in patients with or without anastomotic leakage. PMID- 15521938 TI - Long-term follow-up of dynamic graciloplasty for faecal incontinence. AB - OBJECTIVE: This paper presents the long-term morbidity, function and quality of life data for patients who have undergone dynamic graciloplasty (DGP) for faecal incontinence. PATIENTS AND METHODS: All patients (n = 38) who had undergone DGP at one institution between 1993 and 2003 are presented. Thirty-three were available for long-term follow-up (median 60 months) and completed a telephone questionnaire assessing quality of life (QOL), bowel and sexual function and patient satisfaction. All patients had interval anorectal physiology studies. RESULTS: At a median follow-up of 5 years, 72% had pain, swelling or paraesthesia in the donor leg and 27% had sexual dysfunction. Sixteen percent of patients had been converted to an end-colostomy for persisting incontinence and 11% for obstructed defaecation. All other patients have a normally functioning graciloplasty. Sixteen percent of patients reported a faecal continence score < 12. Of those patients with a functioning graciloplasty, 50% had obstructed defaecation and 64% reported that their bowel dysfunction had a negative impacted on their QOL. Age, medical comorbidity and anal manometry did not correlate with functional outcome. Quality of life scores and patient satisfaction scores correlated significantly with continence scores. There was a trend toward higher QOL and satisfaction scores with conversion to colostomy compared with a continence score > 12. Sixty percent of patients rated their satisfaction with DGP as 50% or better on a visual analogue scale, and this correlated strongly with the continence score at the time of the assessment (P < 0.001). CONCLUSION: Dynamic graciloplasty significantly improves patient quality of life and anal continence for some patients. Despite increased experience, morbidity remains high and long-term continence scores are poor in a majority of cases. Obstructed defaecation is a significant problem after graciloplasty and antegrade colonic enemas may be needed. Significant prognostic factors for obstructed defaecation remain to be identified. The mechanism of both continence failure and surgical morbidity remains poorly defined in many patients and requires further investigation. The individual patient can expect a 16% chance of normal faecal continence at 5 years, with at least one surgical morbidity as a result of the procedure. PMID- 15521939 TI - Shorter hospital stay associated with fastrack postoperative care pathways and laparoscopic intestinal resection are not associated with increased physical activity. AB - OBJECTIVE: Recent prospective studies have shown that 'fast track' postoperative care protocols (FT) can reduce hospitalization after major intestinal surgery to 4.5 days, as compared to the 7-10 days with traditional management (TR) and 2.5 days after laparoscopic surgery (LC). We used computerized actigraphy (CA) to evaluate physical activity using TR, FT and LC approaches. METHODS: Fifteen patients undergoing intestinal resection by LC, TR and FT were recruited. CA devices were placed on the wrist and ankle of each patient from day 1 until hospital discharge. Intra-patient and intergroup analyses were performed using 24 h evaluations, and periods from 0800-2000 h. Results are presented for, level of activity (LOA; activity counts per minute), activity index (AI;% epochs with activity) and acceleration index (ACI; change in activity rate during analysis period) for 0800-2000 h. RESULTS: Levels of activity measured by wrist and ankle CA devices for the three different care protocols were: LC, 6263 +/- 8008 (wrist), 1640 +/- 3795 (ankle); FT 7874 +/- 8550 (wrist), 2153 +/- 4838 (ankle); LC 8526 +/- 9917 (wrist), 2326 +/- 4905 (ankle). Length of hospital stay was significantly shorter in LC than FT and TR patients (P < 0.01). There was no significant increase in wrist or ankle LOA for FT or LC patients. Similarly, there was no difference in AI or ACI. CONCLUSION: Although CA is an easily standardized method of evaluating physical activity after major abdominal surgery, no difference can be demonstrated between activity levels in patients managed by differing care pathways. Reductions in stay may be associated with factors other than a change in the level of physical activity after surgery. PMID- 15521940 TI - Effects of ovariectomy and hormone replacement on submucosal collagen and blood vessels of the anal canal of rats. AB - OBJECTIVE: To study the effects of oestrogen and progesterone on submucosal collagen fibres and vascular plexus of the anal canal. MATERIALS AND METHODS: Experiments were performed on sections of the anal canal of ovariectomized rats following 28 daily subcutaneous injections of 17-beta oestradiol (n = 6, OVX + E, Group 1), medroxyprogesterone acetate (n = 6, OVX + P, Group 2), both drugs (n = 6, OVX + E + P, Group 3) or vehicle (n = 6, OVX) and after sham surgery without castration or injection (n = 6). Investigations included immunohistochemistry of oestrogen and progesterone receptors and collagen fibres, Western blot analysis of collagen types I and III and counting of perianal vessels by light microscopy. RESULTS: There was positive immunostaining for oestrogen and progesterone receptors in the mucosa and for collagen types I and III in the submucosa in all samples. Type I collagen levels increased significantly with ovariectomy but were normalized with treatment with oestrogen and progesterone. Type III collagen levels decreased after ovariectomy. Administration of oestrogen and progesterone appeared to restore level to near sham values. Semi-quantitative measurement of Type I/III collagen ratios by signal intensity demonstrated a very high ratio after ovariectomy. This appeared to be restored by both oestrogen and progesterone administration either individually or in combination. Mean vessel count was significantly lower in sham animals compared to values in OVX animals (P = 0.006). However, while only oestrogen treatment increased significantly the number of vessels compared to sham animals (P = 0.04), replacement with progesterone did not affect and in combination with oestrogen reduced submucosal vessel number. CONCLUSION: Oestrogen and progesterone have synergistic effects on collagen types I and III and probably antagonistic effects on the vascular plexus of the anal canal submucosa in adult female rats. PMID- 15521941 TI - Prospective follow-up after ambulatory plain midline excision of pilonidal sinus and primary suture under local anaesthesia--efficient, sufficient, and persistent. AB - OBJECTIVE: Optimal surgical treatment for chronic pilonidal sinus (PS) disease should be easy and efficient. The purpose of this study was to establish the feasibility of and results after ambulatory simple midline excision and primary wound closure under local anaesthesia. PATIENTS AND METHODS: The prospective nonrandomised single institution study included 131 consecutive PS patients admitted for surgery. After standardized surgery, all patients were prospectively followed until fully healed. Complications were registered. Results after 41 months (mean, range 36-59) were evaluated by questionnaire. RESULTS: 95% were primary closed. All were ambulatory treated and 98% under local anaesthesia. 88% healed in two weeks. 5.6% exhibited minor wound healing defects and 6.4% demanded full debridement of the wound. Mean delayed healing time was 7.5 weeks (range 3 12). 8% developed a recurrence. 10% of the remainders reported a slight tenderness in the scar area. 90% were symptom-free. CONCLUSION: PS treatment as of above is readily achieved. Wound related complications were reasonably few and in the same magnitude as after more complex excision techniques and induced no significant long-term sequels. Recurrences could, however, be more common. Prospective randomised studies are warranted to determine if there are any significant differences between excision techniques in and out of the midline. PMID- 15521942 TI - The risk of dysplasia and cancer in the ileal pouch mucosa after restorative proctocolectomy for ulcerative proctocolitis is low: a long-term term follow-up study. AB - AIM: Some of the rare complications reported in patients with an ileopouch anal anastomosis (IPAA) after coloectomy for chronic ulcerative colitis are dysplasia and carcinoma. The supposed pathway is for the ileal pouch mucosa to go through adaptational changes then is to progress through the phases of chronic pouchitis, dysplasia and subsequently to adenocarcinoma. In many of these studies however, the dysplasia-cancer sequence is inconclusive since the carcinoma might have developed from the ileal mucosa itself or from residual viable rectal mucosa left behind. The purpose of this study was therefore to study the long-term ileal mucosal adaptation patterns and the incidence and grading of dysplasia in the ileal pouch mucosa in patients previously operated on for ulcerative proctocolitis. PATIENTS AND METHODS: Forty-five patients who had been operated on with an IPAA (25 males/20 females), with a median age of 54 years (range 34-76), were invited for clinical examination and pouch endoscopy including mucosal biopsies. The duration of their colitis until surgery was median 6 years (range 1 28) and the time median interval from start of disease until time of follow up 24.8 years (range 17-46). Three independent pathologists from two different centres reviewed sequential mucosal biopsies taken from separate sites of the pouch for dysplasia and mucosal adaptation patterns. RESULTS: The type C pattern with a severe inflammation in lamina propria together with severe atrophy of villi, sometimes with ulceration and granulation tissue, was observed by the two pathologists from one centre in 15 of 45 (33.3%) patients and in 11 (24.4%) of 45 by the third pathologist, respectively. As regards dysplasia one pathologist group evaluated 2/45 (4.4%) cases as low-grade dysplasia while the third pathologist considered one of these cases as indefinite for dysplasia and one as reactive. There was in this respect full agreement between the two centres in 43 (95.6%) of 45 cases. Neither high-grade dysplasia nor invasive carcinoma was diagnosed. CONCLUSION: Dysplastic transformation within the ileal pouch mucosa in patients operated for ulcerative proctocolitis is rare even after a long follow up. These results are reassuring for both patients and surgeons. There seem to be no solid grounds to support routine surveillance for dysplasia in the ileal pouch mucosa in these patients. The surveillance for neoplastic changes in the remaining muscular/epithelial cuff is a separate issue however. PMID- 15521943 TI - Scintigraphic assessment of slow transit constipation with special reference to right- or left-sided colonic delay. AB - OBJECTIVE: Subtotal colectomy and ileorectal anastomosis for slow transit constipation has several side-effects. The motor abnormality in some patients may be segmental which could motivate a limited resection of the colon. Therefore a diagnostic tool to identify a segmental colonic motor dysfunction is needed. The aim of this study was to evaluate a scintigraphic method to assess colonic transit with special reference to right- or left-sided delay. METHODS: Twenty three constipated patients (19 women, mean age 50 years) with slow colonic transit on radio-opaque marker studies and 13 healthy individuals (11 women, mean age 46 years) were studied. All subjects were examined with oral (111)Indium-DTPA scintigraphy. The scintigraphic results for patients and controls were presented as geometric centre of radioactivity and percent activity over time in the right, the left and the recto-sigmoid colon. The inter-observer variation in the interpretation of the scans was also evaluated. RESULTS: There was no difference in transit time between the groups of patients and controls in the right colon whereas the patients had a significant delay in the left colon (P < 0.05). Two patients had a marked delay in the right colon followed by relatively rapid transit in the left colon. The inter-observer correlation was good comparing the right, the left and the recto-sigmoid colon (r = 0.58-0.98, P < 0.01-0.001). CONCLUSION: The results indicate that colonic scintigraphy with oral (111)Indium DTPA may help to select patients for a left or, in a few cases, a right hemicolectomy for slow transit constipation. PMID- 15521944 TI - Colorectal surgery: the risk and burden of adhesion-related complications. AB - OBJECTIVES: Adhesions are associated with serious medical complications. This study examines the real-time burden of adhesion-related readmissions following colorectal surgery and assesses the impact of previous surgery on adhesion related outcomes. PATIENTS AND METHODS: The study used data from the Scottish National Health Service Medical Record Linkage Database to identify three cohorts of patients who had undergone open colorectal surgery during the financial years 1996-97, 1997-98 and 1998-99. Each cohort was followed up for at least 2 years and the number and category of adhesion-related readmissions was recorded. The influence of any previous operations on adhesion-related readmissions was also determined by performing a subanalysis within the 1996-97 cohort of patients who had no record of abdominal surgery within either the previous 5 or 15 years. The relative risk of adhesion-related readmissions was also assessed. RESULTS: In the 1996-97 cohort, 9.0% of patients were readmitted within a year after surgery; 2.1% had complications directly related to adhesions and 6.9% had complications that were possibly related. After 4 years, 19.0% of patients were readmitted for reasons directly or possibly related to adhesions. Many patients were readmitted on more than one occasion and the relative risk of adhesion-related complications was 29.7 per 100 initial procedures over 4 years. In the subgroups that had no record of abdominal surgery within the previous 5 or 15 years, the relative risks of adhesion-related complications were 24.8% and 23.5%, respectively. There was no change in the rate of adhesion-related readmissions following colorectal surgery between 1996 and 1999. CONCLUSION: Colorectal surgery is associated with a considerable rate of adhesion-related readmissions. Preventative measures should be considered to reduce this risk. PMID- 15521945 TI - Absence of socioeconomic variation in survival from colorectal cancer in patients receiving surgical treatment in one health district: cohort study. AB - OBJECTIVE: To examine whether there is an association between patient deprivation status and survival from colorectal cancer among patients receiving treatment of the same type and quality. PATIENTS AND METHODS: A survival study was conducted of all colorectal cancer patients diagnosed between 1991 and 1997 who received surgery either in the NHS district general hospital or the private hospital of one UK health district. The five-year survival rates, both all cause and colorectal cancer specific, were calculated for subgroups defined by patient age, gender, stage and deprivation status using Kaplan-Meier curves. Cox proportional hazards models were used to examine the influence of deprivation on five-year survival after adjusting for age, gender and stage. RESULTS: There were 603 consecutive colorectal patients during the study period. Five-year all-cause and colorectal cancer-specific survival rates were 41% and 53%, respectively. There was no association between deprivation status and stage at diagnosis (P = 0.308). Multivariable proportional hazards modelling (adjusting for gender, age and tumour stage) demonstrated no association between deprivation status and survival. CONCLUSION: In this single district study, no relationship between patient socioeconomic status and survival from colorectal cancer could be demonstrated. Consistency in the type and quality of treatment offered to patients by the same clinical teams may have been responsible for the equitable survival outcomes. PMID- 15521946 TI - Delayed stoma formation in Fournier's gangrene. AB - Fournier's gangrene is traditionally treated with prompt surgical debridement and in many cases a diverting colostomy is also fashioned during the same procedure. We present four cases where stoma formation was delayed until the second look procedure. The physiological states at the time of either procedure were compared using POSSUM. The results showed an improvement in the physiological condition in all patients at the time of the second operation, suggesting that a delay can potentially improve prognosis in such cases. PMID- 15521947 TI - What is the Government doing to improve outcomes for patients with bowel cancer? PMID- 15521949 TI - Long-term faecal incontinence following the use of botulinum toxin. PMID- 15521951 TI - Nicorandil associated anal ulceration. PMID- 15521953 TI - Molecular detection of thyroid cancer: an update. PMID- 15521954 TI - Radiosurgery for pituitary adenomas. PMID- 15521955 TI - Maternal soluble tumour necrosis factor receptor type 2 (sTNFR2) and adiponectin are both related to blood pressure during gestation and infant's birthweight. AB - OBJECTIVE: Tumour necrosis factor alpha (TNF-alpha) and adiponectin are strongly related to insulin sensitivity; insulin resistance of pregnancy is a major determinant of infant's birthweight. We aimed to study the contributions of maternal serum concentrations of soluble TNF-alpha receptors (sTNFR1 and sTNFR2) and adiponectin to infant's birthweight. DESIGN: Cross-sectional, hospital-based study of insulin sensitivity during gestation. PATIENTS: Fifty-one healthy women with uncomplicated pregnancy and delivery (except for elective Caesarian section) and their healthy newborn infants. measurements Maternal blood levels of glucose, insulin, glycosylated haemaglobin (HbA1c), sTNFR1, sTNFR2 and adiponectin at delivery; cord-blood levels of sTNFR1, sTNFR2 and adiponectin. RESULTS: At delivery, maternal sTNFR2 correlated with systolic blood pressure (SBP; r = 0.38, P = 0.005). In multiple regression analyses, SBP and HbA1c were independent predictors of sTNFR2, explaining 18 and 7% of its variance, respectively; insulin resistance index (HOMA-IR), body mass index at delivery and SBP were independent predictors of adiponectin, explaining 15, 8 and 7% of its variance, respectively. Both maternal sTNFR2 and SBP were negatively correlated with infant's birthweight (r = -0.28, P = 0.04 and r = -0.36, P = 0.01 respectively, adjusted for sex and gestational age). In multivariate regression analyses, infant's sex and either maternal sTNFR2 or adiponectin were independent predictors of infant's birthweight, each explaining between 6 and 9% of birthweight variance. Further addition of maternal SBP to these models revealed that this variable was the main predictor of infant's birthweight, explaining 13% of its variance. CONCLUSIONS: Maternal sTNFR2 and adiponectin are independently related to both maternal blood pressure and infant's birthweight in uncomplicated pregnancy. The contributions of the TNF-alpha system and adiponectin to hypertensive disorders of pregnancy and fetal growth merit further studies. PMID- 15521956 TI - Clinical features, diagnosis, treatment and molecular studies in paediatric Cushing's syndrome due to primary nodular adrenocortical hyperplasia. AB - BACKGROUND: Primary nodular adrenocortical hyperplasia (PNAH) is a well recognized, but infrequently studied cause of paediatric Cushing's syndrome (CS). OBJECTIVE: To assess presentation, diagnosis, radiological imaging, treatment and molecular analysis of patients with childhood-onset CS due to PNAH. PATIENTS: Four males and two females (median age 12.9 years, range 10.9-16.9 years) were studied. RESULTS: All had growth failure (mean height SDS -1.2; range -2.5-0.0), weight gain [mean body mass index (BMI) SDS 3.5; range 2.5-4.6] and clinical virilization, while five had hypertension [mean systolic blood pressure (SBP) 130 mmHg, diastolic blood pressure (DBP) 83 mmHg]. One patient had generalized lentigines, one had a tibial chondromyxomatous cyst and two had facial freckling. One patient had a family history of primary nodular adrenocortical disease. The diagnosis of CS was based on elevation of sleeping midnight serum cortisol and urinary free cortisol excretion, and impaired suppression of cortisol on both low and high-dose dexamethasone suppression tests (DST). All patients had undetectable plasma ACTH with absent responses of both plasma ACTH and serum cortisol to an intravenous (i.v.) corticotrophin-releasing hormone (CRH) test. Computed tomography or magnetic resonance imaging showed normal or small adrenals, with nodules in two patients. All patients underwent bilateral adrenalectomy, performed by open (n = 2) or laparoscopic surgery (n = 4) at a mean of 0.4 years (range 0.2-0.8 years) from diagnosis. Hypercortisolaemia was treated preoperatively by metyrapone alone 0.50-0.75 g/day (n = 4), metyrapone 0.75-1.50 g/day + o'p'DDD/mitotane 1-2 g/day (n = 1), or ketoconazole (n = 1). Adrenal histology showed nodular cortical hyperplasia with shrinkage of intervening cortical tissue and pigmentation, present in four patients. Molecular analysis of the type 1-alpha regulatory subunit of protein kinase A (PRKAR1A) gene revealed a novel germline mutation in one patient. Postadrenalectomy, three patients, had catch-up growth with height velocities increasing from 3.0, 3.9 and 2.5-8.9, 8.3 and 9.0 cm/years, respectively. All six are well at a follow-up (mean 4.0 years; range 0.5-10.8 years). CONCLUSIONS: PNAH was associated with cushingoid features, virilization and hypertension with a lack of cortisol suppression on high DST, undetectable plasma ACTH and absent cortisol and ACTH responses to CRH. Adrenals were normal or small on imaging. PRKAR1A gene analysis may be helpful in the assessment of these patients. PMID- 15521957 TI - Testosterone increases bone mineral density in female-to-male transsexuals: a case series of 15 subjects. AB - OBJECTIVE: Testosterone therapy for osteoporosis has not been studied extensively in women because of its potential to cause virilization. Female-to-male transsexuals are genetic females who suffer from gender dysphoria and thus take supra-physiologic doses of testosterone to change from the female to male phenotype. The aim of this study is to examine the effects of testosterone treatment on the genetic female skeleton. PATIENTS AND DESIGN: A group of 15 female-to-male transsexuals was prospectively enrolled for observation over a 2 year period. The subjects had a mean age of 37.0 +/- 3.0 years. All of the subjects self-administered testosterone esters intramuscularly at a mean dose of 70.7 +/- 4.5 mg weekly. MEASUREMENTS: The subjects had measurements of bone mineral density (BMD) by dual X-ray absorptiometry (DXA) of the femoral neck and spine (L2-L4) at 12-month intervals. They had determinations of serum oestradiol, testosterone, soluble RANKL (sRANKL), osteoprotegerin (OPG) and urine N telopeptide (NTX) at the date of enrolment and at the end of 2 years. results There was a significant positive increase in mean BMD of 7.8% at the femoral neck and a nonsignificant increase in mean BMD of 3.1% at the spine over 2 years. The levels of testosterone reached the upper normal range for males and the levels of oestradiol declined to near the postmenopausal range. sRANKL levels decreased significantly in female-to-male transsexuals who newly initiated testosterone therapy. There was no significant change in urine NTX or serum OPG during the study. CONCLUSIONS: We conclude that supra-physiologic testosterone therapy increases BMD at the hip while maintaining BMD at the spine in female-to-male transsexuals. The effects of testosterone may be the result of testosterone hormone directly acting on the bone or indirectly through aromatization to oestradiol. Lower RANKL levels coupled with unchanged OPG levels results in an increased OPG/RANKL ratio, which may be beneficial to the bone by inhibiting osteoclastogenesis. PMID- 15521958 TI - Uterus size and ovarian morphology in women with isolated growth hormone deficiency, hypogonadotrophic hypogonadism and hypopituitarism. AB - OBJECTIVE AND SUBJECTS: Current data suggest a permissive role for the somatotrophic axis in the reproductive process, mainly by affecting the onset of puberty and the maintenance of regular menstrual cyclicity. To assess a possible interface between GH and reproductive axes in determining the uterus size, we retrospectively evaluated 58 pelvic ultrasound scans in adult women with either isolated growth hormone deficiency (GHD; n = 12), hypogonadotrophic hypogonadism (HH; n = 24) or hypopituitarism (HP; n = 22) of prepubertal onset. Pelvic ultrasound was performed before oestrogen replacement in patients with HH or HP, and after completion of GH treatment in the majority of patients with HP. Eight women with HH and seven with HP had a second pelvic ultrasound examination after being established on oestrogen replacement therapy. A group of 19 young healthy women with no previous history of pregnancy or miscarriage were included for comparison of ultrasound data. RESULTS: Uterine measurements, length and uterine cross-sectional area (UXA) were significantly less in the three study groups compared to healthy controls [median UXA: GHD 18.0 cm(2) (range 9.9-28.6 cm(2)), HH 7.0 cm(2) (range 1.3-18.5 cm(2)), HP 11.8 cm(2) (range 1.6-21.8 cm(2)) and controls 23.0 cm(2) (range 16.1-31.7 cm(2)); P < 0.001]. The median age of oestrogen replacement was significantly later in HH [19 years (range 16-26 years)] compared to HP [16.5 years (range 13-20 years)]P = 0.03, while the median age of menarche of GHD patients was 14.5 years (range 11-16 years), which was not statistically different from controls [13.0 years (range 12.5-14.5 years)]. Repeat uterine measurements on oestrogen in the subgroup of 15 patients did not reach the normal values [pretreatment UXA: 4.2 cm(2) (range 1.6-16.1 cm(2)), post treatment UXA: 12.6 cm(2) (range 4-23 cm(2))]. Ovarian volume was smaller in the two groups of women with gonadotrophin deficiency (HH and HP), while a polycystic ovarian morphology was notably more prevalent in the two groups who had received GH treatment being found in 75 and 58% of women with GHD and HP, respectively, compared with 12.5% in women with HH (P < 0.004). CONCLUSIONS: Our findings suggest that GHD might have an independent effect in determining uterus size and therefore the consequences of GHD plus oestrogen deficiency on uterus growth might be additive. The fact that uterine measurements between HH and HP patients did not differed significantly in this study may be explained by the fact that oestrogen replacement has been substituted earlier in the latter group. Furthermore, it appears that standard oestrogen replacement therapy did not result in normal uterine growth. A polycystic ovarian morphology may be a consequence of GH treatment. PMID- 15521959 TI - Characterization of a promoter polymorphism in the glucocorticoid receptor gene and its relationship to three other polymorphisms. AB - OBJECTIVE: Sensitivity to glucocorticoids within the normal population is highly variable and partly determined by polymorphisms in the glucocorticoid receptor (GR) gene (NR3C1). We investigated the exact sequence alteration of a TthIIII polymorphism in the GR gene, whether it is associated with glucocorticoid sensitivity, and its relationship to 3 polymorphisms of the GR gene (N363S, BclI, ER22/23EK). DESIGN: Two dexamethasone (DEX) suppression tests were performed with 1 and 0.25 mg DEX, respectively. PATIENTS: We genotyped a random subgroup of 209 participants of the Rotterdam Study, a population-based study in the elderly. MEASUREMENTS: Anthropometric parameters, cortisol, insulin and glucose levels, and lipid concentrations were measured. RESULTS: We identified the TthIIII polymorphism as a C to T mutation, 3807 bp upstream from the mRNA start site. We found 39.7% CC-carriers, 44.5% CT-carriers, and 15.8 % TT-carriers. No differences were found between TthIIII genotypes in sensitivity to DEX, baseline cortisol, insulin, glucose or cholesterol levels, or in anthropometric variables. However, all ER22/23EK-carriers also carried the TthIIII T-allele, and carriers of both these polymorphisms had a significantly smaller cortisol suppression after 1 mg DEX, lower fasting insulin levels, and lower total and low-density lipoprotein (LDL) cholesterol levels than TthIIII T carriers without the ER22/23EK variant and noncarriers. No interaction was found between the TthIIII variant and N363S or BclI polymorphisms. CONCLUSIONS: The TthIIII polymorphism is not functional by itself. However, the ER22/23EK polymorphism is without exception linked to the TthIIII T polymorphism and this haplotype is associated with a relative resistance to glucocorticoids, and a healthy metabolic profile. PMID- 15521960 TI - Responses to the 35% CO challenge in postpartum women. AB - Lactation has been associated with suppression of some components of the neuroendocrine stress response. In humans, suppression of the hypothalamo pituitary-adrenal (HPA) axis has been demonstrated in response to both a psychological and an exercise challenge, but appears to be limited to a short period of time following suckling. Information regarding other components of the stress response and to other challenges in humans is limited. We have evaluated the endocrine, autonomic and psychological response to a single breath of 35% CO(2) during lactation. The 35% CO(2) challenge is a safe and simple test that has been shown to stimulate the HPA axis, produce autonomic activation and emotional arousal. Eight breastfeeding and six bottle-feeding mothers, 6 weeks' postpartum, and eight control women were studied. Twenty minutes following the cessation of feeding, plasma cortisol levels were significantly reduced in the breastfeeding women (P = 0.002 compared with control and P = 0.003 compared with bottle-feeders). Despite this, cortisol, blood pressure, heart rate and psychological responses to the challenge were no different in the breastfeeding group compared to either the control or bottle feeding groups. These results confirm that suckling is associated with short-term suppression of cortisol, but this has no effect on the ability of the mother to mount a normal hormonal, autonomic and psychological response to the 35% CO(2) challenge. PMID- 15521961 TI - Spontaneous reporting of adverse reactions to carbimazole and propylthiouracil in the UK. AB - OBJECTIVE: To determine the frequency and spectrum of serious adverse drug reactions (ADRs) to thionamide antithyroid drugs. DESIGN AND METHODS: Data spontaneously reported to the UK-wide pharmacovigilance agency, The Committee on Safety of Medicines (Yellow Card Scheme), between 1963 and 2003 were analysed to determine the spectrum and relative frequency of ADRs to carbimazole and propylthiouracil. Representative data on the number of dispensed prescriptions were available from 1981, and were used as a denominator to estimate relative reporting rates of ADRs attributed to the drugs. RESULTS: Between 1981 and 2003 there were 5.23 million prescriptions for thionamide drugs in England and Scotland, 94% of which were for carbimazole. Neutrophil dyscrasia (agranulocytosis and neutropenia) accounted for 49% of all fatalities attributed to these medications. The median time reported for the appearance of neutrophil dyscrasia was 30 days of treatment, but there was a wide range (7-875 days). Neutrophil dyscrasia was more frequently fatal in subjects over 65 years of age; 13.8%vs. 1.2% of fatal reports in younger subjects [odds ratio (OR) 12.90; 5-95% confidence intervals (CI) 1.45-114.92]. Since 1981, reports of most ADRs, including neutrophil dyscrasia, were made significantly more frequently per propylthiouracil prescription dispensed than per carbimazole prescription. CONCLUSION: Detailed data about ADRs to these compounds, and their patient demography, based on a large number of reports are described. Neutrophil dyscrasia is the commonest life-threatening ADR to thionamides. The excess of ADR reports for propylthiouracil compared to carbimazole could reflect genuine differences in toxicity between the compounds, the relative unfamiliarity of UK physicians with propylthiouracil, or the higher use of propylthiouracil in certain patient groups. PMID- 15521962 TI - Associations of insulin-like growth factors, insulin-like growth factor binding proteins and acid-labile subunit with coronary heart disease. AB - OBJECTIVE: IGFs and their binding proteins (IGFBPs) are produced both systemically and locally by cells of the cardiovascular system. As growth promoters, they may play a role in atherosclerosis. DESIGN: Case-control, cross sectional. PATIENTS: A total of 95 nondiabetic male patients with coronary heart disease (CHD) and 92 probands from the Prospective Cardiovascular Munster (PROCAM) who were below the age of 60 years and matched by age, body mass index (BMI) and smoking habits. MEASUREMENTS: We analysed the strength and independence of associations of angiographically assessed presence of CHD with BMI, systolic and diastolic blood pressure, total, high-density lipoprotein (HDL) and LDL cholesterol, triglycerides, lipoprotein(a), apolipoproteins A-I and B, total and free IGF-I, IGF-II, IGFBP-1, IGFBP-3, IGFBP-5, acid-labile subunit (ALS), insulin, C-peptide, testosterone, DHEAS and sex hormone binding globulin. RESULTS: Using multivariate statistical analysis, the presence of CHD had significant positive associations with total IGF-I, IGFBP-5, ALS and IGFBP-3. These associations were independent of each other as well as of traditional risk factors, insulin and sex hormones. CONCLUSION: These observations may indicate a pathogenetic role of the GH/IGF axis in coronary atherosclerosis. PMID- 15521963 TI - Failure to normalize parathyroid hormone during treatment of vitamin D deficiency in Asian patients. AB - OBJECTIVE: Vitamin D deficiency and osteomalacia remain commonplace within the Asian community in Bradford. The treatment of vitamin D deficiency and osteomalacia is cheap and effective, but there are few data on long-term outcomes. Studies have suggested that a minority of patients fail to normalize parathyroid hormone (PTH) levels during therapy with vitamin D. This study aimed to determine what proportion of Asian patients with vitamin D deficiency and secondary hyperparathyroidism normalize PTH levels following therapy with oral vitamin D and to examine reasons for failure to normalize PTH. DESIGN: This study examined the impact of an oral regimen of vitamin D 800 i.u. (20 micrograms) and calcium 1000 mg daily, on PTH levels within an endocrinology outpatient clinic. patients 51 (4M:47F) Asian patients, median age 39 years (range 16-77 years) with vitamin D deficiency (25-hydroxyvitamin D < 25 nmol/l) and secondary hyperparathyroidism (PTH > 5.7 pmol/l). MEASUREMENTS: All patients had at least one follow-up measurement of PTH and calcium during treatment. A subgroup of patients gave consent for examination of GP-prescribing records to indirectly asses adherence to therapy. RESULTS: PTH normalized in only 28/51 (55%) patients (group N) and failed to normalize in 23/51 (45%) patients (group F). Baseline patient characteristics including: age, basal serum 25-hydroxyvitamin D (25OHD), basal serum PTH, basal serum calcium and post treatment serum calcium, were similar in groups N and F. Mild hypercalcaemia occurred in only two (3.9%) patients. The proportion of prescriptions collected by patients in group N was 75 (17-100)% and in group F was 17 (0-100)%, P < 0.0001. CONCLUSIONS: This study has demonstrated that long-term oral treatment with vitamin D and calcium, fails to normalize PTH in a significant proportion of patients with vitamin D deficiency and osteomalacia. This is most likely related to lack of adherence to long-term treatment. Improved ways of treating this condition need to be explored. PMID- 15521964 TI - Serum homocysteine concentrations in children with growth hormone (GH) deficiency before and after 12 months GH replacement. AB - OBJECTIVE: This open, prospective study was designed to evaluate the effect of growth hormone deficiency (GHD) and GH replacement therapy on serum homocysteine (Hcy) concentration in children with GHD. SUBJECTS: Seventeen prepubertal children with GHD (11 boys and six girls) aged 8.6 +/- 1.9 years were studied before and after 12 months of GH replacement therapy at a dose of GH of 30 microg/kg/day. Seventeen healthy children acted as controls and were matched for age, sex and body mass index (BMI). METHODS: At study entry, height, weight, blood pressure, serum Hcy, serum IGF-I, total-low density lipoprotein (LDL)- and high density lipoprotein (HDL) cholesterol, triglycerides, free T4, free T3, vitamin B12, folate, glucose and creatinine were measured in all subjects. The atherogenic index (AI) was also calculated as the ratio of total cholesterol/HDL cholesterol (T/HDL). In GHD children these parameters were also revaluated after 12 months of GH therapy. RESULTS: At study entry height and serum IGF-I were significantly lower, as expected, in GHD patients than in controls (P < 0.0001 and P < 0.007, respectively). Serum Hcy levels were significantly higher in GHD patients than in healthy children (8.4 +/- 2.9 vs. 6.0 +/- 2.9 micromol/l; P < 0.03), although the absolute values were within the normal values for age and sex. There were no significant differences at baseline with respect to blood pressure, serum vitamin B12, folate, fT3, fT4, lipid profile, creatinine and glucose levels. After 12 months of GH replacement therapy height and serum IGF-I increased significantly compared to pretreatment values (P < 0.0001); serum Hcy levels decreased significantly (6.0 +/- 3.3 micromol/l; P < 0.002) compared to baseline values, becoming similar to control values. Total cholesterol (3.5 +/- 0.6 mmol/l) and the AI (2.5 +/- 0.8) decreased significantly with respect to both pretreatment (4.2 +/- 1.0 mmol/l; P < 0.0002 and 3.4 +/- 0.8; < 0.002, respectively) and control values (4.2 +/- 0.4 mmol/l; P < 0.0005 and 3.3 +/- 1.1; P = 0.02, respectively). CONCLUSIONS: GHD in children is associated with higher serum levels of Hcy compared to controls, without significantly affecting the lipid profile. GH replacement for 12 months significantly decreased the Hcy levels and improved the lipid profile with a decrease of total cholesterol and the total/HDL cholesterol ratio, compared to pretreatment values. Given the small number of patients, further larger studies are needed to clarify whether these results may have significant effects in the prevention of cardiovascular disease in adulthood. PMID- 15521965 TI - Clinical results of anti-inflammatory therapy in Graves' ophthalmopathy and association with thyroidal autoantibodies. AB - OBJECTIVE: Graves' ophthalmopathy (GO) is clinically associated with autoimmune thyroid disease, and autoantibodies to thyroidal antigens, especially to the TSH receptor (TRAb), might be involved in the disease process. While there is mounting evidence that TRAb are associated with GO at the onset of the disease, so far no studies have looked at the association between thyroidal autoantibodies and the clinical outcome of GO therapy. The aim of this retrospective study was to evaluate whether TSH binding inhibitory immunoglobulins (TBII) and thyroid stimulating antibodies (TSAb) are still associated with the clinical activity and severity of GO after the completion of anti-inflammatory therapy. In addition, we wanted to elucidate whether thyroid peroxidase (TPO) or thyroglobulin (TG) autoantibodies (TPOAb and TGAb) are in any way related to GO. DESIGN PATIENTS AND MEASUREMENTS: Clinical activity score (CAS) and the severity of GO (modified NOSPECS score) were assessed in 108 patients with GO after steroid therapy and, if indicated, orbital irradiation. Patients were grouped according to their clinical presentation and autoantibody levels (TBII, TSAb, TPOAb and TGAb) were measured. After therapy for hyperthyroidism, all patients were clinically euthyroid but showed clear heterogeneity for GO 4-12 months after the end of anti inflammatory therapy. Fifty-two patients had inactive GO, 41 had moderately active and 15 still had very active (non-responsive) GO. Concerning severity, 27 patients had mild GO, 64 moderately severe and 17 severe GO. RESULTS: TBII titres were still positive in 14 (93%) of 15 patients in the non-responsive group (CAS > 6) compared to 22 (42%) of 52 patients (P < 0.001) with post-therapeutic inactive GO (CAS 0.5, but FICI < or = 4) or antagonistic (FICI > 4). The fungicidal activity of nikkomycin Z alone and in combination with a representative echinocandin (caspofungin) or triazole (voriconazole) was also examined with time kill experiments and fungal cell viability assays. Two-drug combinations of nikkomycin Z with amphotericin B (FICI 3.59 +/- 0.57), amphotericin B lipid complex (FICI 3.95 +/- 0.74), liposomal amphotericin B (FICI 3.62 +/- 0.98), itraconazole (FICI 2.0 +/- 0.0), voriconazole (FICI 1.07 +/- 0.37), posaconazole (FICI 2.20 +/- 0.44) or ravuconazole (FICI 1.76 +/- 0.44) showed no interactions, but the pairwise combination of nikkomycin Z with caspofungin (FICI 0.22 +/- 0.19) or micafungin (FICI 0.35 +/- 0.27) showed synergic activity against A. fumigatus. Time-kill studies and fungal cell viability assays showed that neither nikkomycin Z nor caspofungin alone possessed fungicidal activity against A. fumigatus, whereas a combination of these two drugs at concentrations > or = 2 mg/L (> or = 0.031 x the concentration of drug that produced no visible growth) killed germinated conidia within 24 h in a concentration-dependent manner. These data suggest that two-drug combinations of nikkomycin Z with echinocandins, but not with polyenes and triazoles, have a synergic effect against A. fumigatus. PMID- 15521998 TI - Prospective evaluation of a dot-blot enzyme immunoassay (Directigen RSV) for the antigenic detection of respiratory syncytial virus from nasopharyngeal aspirates of paediatric patients. AB - This study investigated the efficacy of a commercial enzyme immunoassay (Directigen RSV, ColorPAC) in comparison with the shell vial culture method (using Hep-2 cells) for the detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates from children with bronchiolitis. During the period 1995 2002, 4950 samples were examined. RSV was detected in 1660 (33.5%) samples, with a sensitivity of 80.9%, a specificity of 97.5%, a positive predictive value of 93.8%, a negative predictive value of 91.6%, and a testing efficiency value of 92.2% compared with shell vial culture. In 83 (5%) samples, the ColorPAC was positive and the shell vial assay was negative. Of these, 71 (85.6%) were false negative by cell culture. The true false-positive results obtained by ColorPAC represented only 0.7% of all RSV-positive samples. In general, no statistically significant differences were detected between the different months and epidemic periods studied. Compared with ColorPAC, the shell vial culture method displayed a sensitivity of 95.8% and a specificity of 100%. Overall, the ColorPAC assay was an acceptable, simple and rapid method for the antigenic detection of RSV in paediatric respiratory samples. PMID- 15521999 TI - Carriage of resistant microorganisms in repatriates from foreign hospitals to The Netherlands. AB - In a prospective survey conducted between May 1998 and September 2001, the prevalence of carriage of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE) and gentamicin-resistant Gram-negative bacilli (GGNB) was determined in 1167 patients repatriated from foreign hospitals to The Netherlands. Swab specimens, demographic data and clinical data were obtained during transfer of the patients from the foreign hospitals. The total prevalence of carriage of resistant microorganisms was 18.2%. MRSA was carried by 2.7% of all patients, and by 4.7% of the patients repatriated to a Dutch hospital. Antimicrobial treatment (adjusted odds ratio (OR) 3.4; 95% confidence interval (CI) 1.2-9.7), length of stay in a foreign hospital of > 14 days (adjusted OR 5.4; 95% CI 2.3-12) and artificial ventilation (adjusted OR 8.5; 95% CI 1.8-41) were risk factors for carriage of MRSA. VRE and GGNB were isolated from 2.7% and 14.1% of the patients, respectively. Transfer from Asia, and southern, southeastern and eastern Europe, were risk factors for carriage of GGNB. These carriage rates were high compared to those found in patients in Dutch hospitals, where the rates are < 1% for MRSA, 2% for VRE, and 4.5% for GGNB. The highest risk of acquisition of GGNB was associated with the country from where the patient was repatriated, rather than with the antimicrobial treatment received by the individual patient in the foreign hospital. PMID- 15522000 TI - The influence of BCG immunisation on tuberculin reactivity and booster effect in adults in a country with a high prevalence of tuberculosis. AB - The relationship of age and previous BCG vaccination with tuberculin skin test (TST) reactivity was investigated to assess the interpretation of TST results in the adult population of Turkey, where there is a high prevalence of tuberculosis and a routine BCG vaccination programme. The influences of age and BCG vaccine status on booster reaction were also evaluated. TST was applied (5 tuberculin units of purified protein derivative intradermally) to two healthy adult groups, namely 98 medical students and 187 elderly people in a retirement home. The TST was considered positive if an induration > or = 10 mm in diameter was produced. Subjects (41 elderly people and 39 students) with a reaction < 10 mm in diameter were retested 1 week later. There was no significant difference between the students (59.1%) and elderly subjects (58.8%) with respect to positive TST response. No influence of BCG scars on TST reactivity was observed in either group. The booster effect was seen more commonly in the elderly, but the presence of a BCG scar did not influence the booster effect in either group. It was concluded that a positive TST response and booster reaction in adults in high prevalence countries may be caused by latent tuberculosis rather than previous vaccination. PMID- 15522001 TI - Assessing the residual antibacterial activity of clinical materials disinfected with glutaraldehyde, o-phthalaldehyde, hydrogen peroxide or 2-bromo-2-nitro-1,3 propanediol by means of a bacterial toxicity assay. AB - This study investigated the use of a rapid bacterial toxicity test for detecting disinfectant residues released by disinfected materials. The test substances included an environmental disinfectant used in hospitals in high-risk areas, such as critical care units or emergency services, and three disinfectants used on clinical devices when a high level of disinfection is required. The test materials were polyurethane, polypropylene, glass, latex and cotton from different instruments and utensils used in hospitals. Of the four test disinfectants, o-phthalaldehyde (OPA) and 2-bromo-2-nitro-1,3-propanediol (BNP) showed the greatest inhibitory activity (as much as 300-fold greater than hydrogen peroxide in the case of OPA) according to the toxicity text. However, with the exception of hydrogen peroxide on latex, it was the most porous test materials, namely latex and cotton, that accumulated the least residue. BNP was the disinfectant that left the least residue on the five test materials, while the greatest residual concentration was left by hydrogen peroxide on latex (as much as 5 microg/cm2). The biotest used in this study permitted the detection of disinfectant residues released by different types of previously disinfected clinical materials, and can be adapted to simulate elution conditions similar to those existing in routine hospital practice. PMID- 15522002 TI - Relationship between pharmacokinetics and pharmacodynamics of beta-lactams and outcome. AB - The in-vitro susceptibility of an organism and the pharmacokinetics of an antimicrobial agent are two basic factors on which the choice of standardised treatment regimens is based. However, the inter-individual variability of these factors, which modifies the exposure of bacteria to an antibiotic in terms of time and quantity, is not usually taken into account. In 87 patients treated with beta-lactams (ceftriaxone, cefepime or piperacillin), the probability of failure was greater when the infectious process was located in tissues with barriers to the distribution of beta-lactams. Mean MICs of piperacillin and cefepime, but not ceftriaxone, were below the breakpoints in cases of both recovery and failure, but organisms isolated from patients with a poor outcome had higher MICs. Therefore, the use of breakpoints to determine the susceptibility of microorganisms was not satisfactory in predicting the outcome for a large number of patients. If MICs are determined and plasma concentrations are monitored, dosages can be adjusted according to these parameters, thereby allowing antibiotic treatment to be individualised. PMID- 15522003 TI - Emergence of vancomycin-resistant enterococci in a tertiary hospital in Crete, Greece: a cluster of cases and prevalence study on intestinal colonisation. AB - The aim of this study was to investigate the clinical and epidemiological characteristics of five consecutive cases of infection with vancomycin-resistant enterococci (VRE) and the prevalence of faecal carriage of VRE among patients admitted to a 700-bed university hospital where no VRE had been isolated previously. In a 2-month period, five consecutive patients infected with VRE were detected. Three VanB+ Enterococcus faecium isolates were obtained from three patients, while two VanA+ E. faecium isolates, one VanA+ Enterococcus faecalis isolate and one VanC1+ Enterococcus gallinarum isolate were obtained from the other two patients. Of 218 faecal specimens from all hospital wards, 41 (18.8%) were found to contain VRE. Forty-two isolates of VRE were obtained, comprising one (2%) E. faecalis, 11 (27%) E. faecium, 24 (57%) E. gallinarum and six (14%) Enterococcus casseliflavus/flavescens. Four isolates carried the vanA gene, eight carried vanB, 24 carried vanC1, and six carried vanC2/C3. Use of glycopeptides, the presence of central venous catheters and renal dialysis all correlated with VRE colonisation. The prevalence rates were among the highest reported in the literature. PMID- 15522004 TI - Analysis of virulence factors in cases of enterococcal endocarditis. AB - Eleven isolates of Enterococcus faecalis causing endocarditis were screened for possible virulence factors with PCR and phenotypic assays. The gene coding for the enterococcal surface protein (esp) was detected in one isolate only, and haemolysin was produced by two isolates. Aggregation substance, biofilm formation and gelatinase were present in seven, nine and eight isolates, respectively. Predisposing factors, particularly hospitalisation and multiple antibiotic therapy, appeared to be more relevant to the development of enterococcal endocarditis following bloodstream infections than the pattern of virulence factors. PMID- 15522005 TI - Influence of rifaximin treatment on the susceptibility of intestinal Gram negative flora and enterococci. AB - The development of rifaximin- and rifampicin-resistant intestinal coliforms was studied in 27 subjects receiving rifaximin for 3 days by plating stool samples on media containing rifaximin 200 mg/L or rifampicin 64 mg/L before treatment (day 0), after treatment was completed (day 3), and after a further 2 days (day 5). The susceptibility of enterococci grown on day 0 and day 3 was also studied in 71 subjects. Significant increases in antimicrobial-resistant coliform flora were not seen in either the rifaximin-treated or the placebo-treated subjects. Enterococci recovered pre- and post-treatment showed similar susceptibilities. Rifaximin did not select for significant resistance in the Gram-negative and Gram positive intestinal flora during therapy. PMID- 15522006 TI - Production by Escherichia coli isolates of siderophore and other virulence factors and their pathogenic role in a cutaneous infection model. AB - Escherichia coli isolates from urinary tract infections (UTIs) (n = 124), extra urinary sites (n = 37) and normal faecal samples (n = 51) were examined for the presence of virulence factors, including siderophores (aerobactin and enterobactin). The proportion of aerobactin producers was significantly higher in UTI (69.4%; p 0.001) and extra-urinary samples (70.3%; p 0.007) than in controls (41.2%), while the proportion of enterobactin producers was significantly lower in the UTI samples than in the controls (p 0.027). In a cutaneous infection model, aerobactin-positive E. coli showed more growth than non-aerobactin and non enterobactin isolates, even when other virulence factors were identical. PMID- 15522007 TI - Mycobacterial testing in hospital laboratories: results from a questionnaire survey in Italy. AB - Between 1999 and 2001, 355 hospital laboratories in Italy were asked to complete a questionnaire addressing mycobacterial test methods, 1-year workloads and laboratory safety features. Analysis of the data showed that rapid methods for mycobacterial testing were being used by most larger laboratories; however, sub optimal methods were still in use in small and medium-size laboratories. In a country such as Italy, which has a low prevalence of tuberculosis cases, implementation of rapid technologies, combined with regionalisation of mycobacterial diagnostic services, seems to be the most reasonable and cost effective strategy. PMID- 15522008 TI - Isolation of a novel sequevar of Mycobacterium flavescens from the synovial fluid of an AIDS patient. AB - This report describes the characterisation of a mycobacterium involved in a case of septic arthritis in an AIDS patient that was treated successfully with specific anti-mycobacterial drugs. The biochemical and cultural features, and the mycolic acid pattern as assessed by high-performance liquid chromatography, were fully compatible with the isolate being Mycobacterium flavescens. However, the isolate's 16S rDNA sequence differed by five nucleotides from the two known sequevars of M. flavescens, thus indicating that this isolate belonged to a new 16S rDNA sequevar. PMID- 15522009 TI - Use of sulesomab in the diagnosis of brucellar spondylitis. AB - Twenty-two patients with suspected brucellar spondylitis were investigated to evaluate the possible diagnostic role of Sulesomab, a (99m)Tc-antigranulocyte antibody Fab' fragment. Sensitivity and specificity were compared with those of magnetic resonance imaging (MRI). Skeletal involvement was detected by MRI in 11 cases, while leukoscintigraphy indicated normal vertebral uptake in seven of these patients, increased uptake in two patients, and decreased uptake in two patients. Leukoscintigraphy of the 11 patients negative by MRI demonstrated increased uptake in two cases. The sensitivity and specificity of leukoscintigraphy were 27.2% and 81.1%, respectively. Based on these results, leukoscintigraphy is not indicated for the management of patients with suspected brucellar spondylitis. PMID- 15522010 TI - Outbreak of echovirus 13 infection among Lithuanian children. AB - The aim of this study was to investigate an outbreak of enterovirus infection in Lithuania in 2001, as well as the clinical presentation and outcome of the disease. Thirty children aged 1 month to 15 years were referred to hospital with suspected enterovirus infection during the period August-October 2001. Echovirus 13 (EV-13) was isolated from eight (26.7%) cases. No other pathogens were detected. Infection presented predominantly as viral meningitis, and rarely as a febrile illness with rash. The outcome was good for all patients. This is the first report of EV-13 from Lithuania. PMID- 15522011 TI - Molecular epidemiology of astrovirus infection in Italian children with gastroenteritis. AB - Abstract A 1-year study involving 157 gastroenteritis samples was conducted to investigate the role of human astrovirus (HAstV) as a cause of gastroenteritis in Italian children aged < 2 years. The overall incidence of HAstV was 3.1%. Most cases occurred between March and May, and four of the five isolates were of the HAstV-1 type, the other being HAstV-3. Analysis of genetic variability showed that the three HAstV-1 isolates collected in 2000 clustered together, but separately from the 1999 isolate. The results indicated that HAstV should be considered as a potential diarrhoeal pathogen in Italian children. PMID- 15522012 TI - High seroprevalence of Pneumocystis infection in Spanish children. AB - Pneumocystis infection occurs worldwide, and most individuals test seropositive for Pneumocystis early in childhood. Little is known about the epidemiology of this infection in western Europe. The seroprevalence of Pneumocystis infection in 233 Spanish children was determined in a community study by immunoblot analysis of sera. The overall seroprevalence was 73%, with an age-related increase from 52% at 6 years to 66% at 10 years and 80% at 13 years. The data indicated a high seroprevalence of Pneumocystis infection in healthy Spanish children, thereby demonstrating that this pathogen is widespread in southern Spain. PMID- 15522013 TI - Salmonella Enteritidis in Bosnia and Herzegovina. AB - A comprehensive retrospective analysis of human Salmonella Enteritidis isolates in the Zenica-Doboj Canton of Bosnia and Herzegovina was conducted by inquiry and questionnaire. In the period 1998-2000, 299 isolates of Salmonella spp. were recorded, of which S. Enteritidis accounted for 74.2%. The isolation rate of S. Enteritidis increased during this period, from 12.7 to 25.5 isolates/year/100,000 population. Isolates were obtained all year round, mostly from sporadic cases of infection or limited family outbreaks. Home-made food was identified as the most important source of infection, being implicated in 81% of outbreaks and 81.7% of cases of sporadic infection. PMID- 15522014 TI - Increasing incidence of resistance to nalidixic acid in shigellas from humans in England and Wales: implications for therapy. AB - Among shigellas isolated from patients in England and Wales in 2002, 10% of subgroups A, B and C, and 13% of subgroup D (Shigella sonnei), were resistant to nalidixic acid. As a consequence, should antimicrobial therapy be indicated, the efficacy of nalidixic acid as the preferred treatment for children with bacillary dysentery has been jeopardised. PMID- 15522016 TI - Patterns of detection of superficial spreading and nodular-type melanoma: a multicenter Italian study. AB - BACKGROUND: Nodular histotype represents the condition that is mostly associated with diagnosis of thick melanoma. OBJECTIVE: The objectives were to evaluate variables associated with and pattern of detection of nodular melanomas and to investigate variables associated with early diagnosis in accordance with histotype (nodular vs. superficial spreading melanomas). METHODS: From the original data set of 816 melanomas, all the invasive lesions classified as superficial spreading (n=500) and nodular (n=93) melanomas were considered for the study. A multivariate logistic analysis was performed. Results. Nodular melanomas did not significantly differ from superficial spreading melanomas regarding sex, anatomic site, number of whole-body nevi, and the presence of atypical nevi. As expected, nodular melanomas were represented by a higher percentage of thick (>2 mm) lesions compared to superficial spreading melanomas (64.5% vs. 9.6%, p<0.001). The pattern of detection significantly differed between nodular and superficial spreading melanomas, the former being more frequently self-detected (44.1% vs. 38.0%) or detected by the family doctor (34.4% vs. 11.4%). Female sex, high level of education, and detection made by a dermatologist had an independent, protective effect against late (>1 mm in thickness) diagnosis in superficial spreading melanomas. No protective variable associated with nodular melanomas was found. CONCLUSION: Patterns of detection for nodular melanomas significantly differ from those for superficial spreading melanomas. For superficial spreading, but not for nodular, melanomas, variables associated with protective effect against late diagnosis can be identified. PMID- 15522017 TI - Why are there differences in the perceived safety of office-based surgery? AB - BACKGROUND: Office-based surgery has become an important method of health-care delivery, but there is controversy about its safety and which practitioners should perform it. Several states have already or are preparing to enact legislation regulating office-based surgery. OBJECTIVE: The objective was to discuss recent literature pertaining to the safety of office surgery and to discuss reasons why there are perceived differences in its safety. METHODS: The pertinent literature is reviewed. Results. The majority of studies suggest that office surgery is safe. A recent study that found to the contrary may have methodologic flaws. CONCLUSION: The medical and legislative community should seek to scientifically examine office surgery. Overregulation or loss of office surgery would have a tremendous impact on the management of skin cancers and the delivery of quality cosmetic and laser surgery. PMID- 15522018 TI - Intravascular 1320-nm laser closure of the great saphenous vein: a 6- to 12-month follow-up study. AB - OBJECTIVE: The objective was to determine the safety and efficacy of an intravascular laser with a novel wavelength to close the great saphenous vein. METHODS: Twenty-four cases of an incompetent great saphenous vein (0.5-1.2 cm in diameter) associated with distal varicose veins were treated with a 1320-nm intravascular laser at 5 W with an automatic pullback mechanism at 1 mm/s. Patients were evaluated with duplex ultrasound to determine efficacy of treatment at various time periods to at least 6 months after the procedure. RESULTS: All patients demonstrated complete closure of the incompetent great saphenous vein. In most cases, the treated great saphenous vein was not identifiable 6 months postoperatively. There was no recurrence of any varicose veins. All preoperative symptoms resolved after treatment, and no complications were noted. All patients were very pleased with the outcome of the procedure. CONCLUSIONS: At 6 months or greater follow-up, a 5-W, 1320-nm intravascular laser with 1 mm/s automatic pullback, delivered through a diffusion-tip fiber, is safe and effective in treating an incompetent great saphenous vein up to 1.2 cm in diameter. PMID- 15522019 TI - Microfoam ultrasound-guided sclerotherapy treatment for varicose veins in a subgroup with diameters at the junction of 10 mm or greater compared with a subgroup of less than 10 mm. AB - OBJECTIVE: The objective was to analyze the effectiveness of foam ultrasound guided sclerotherapy treatment in saphenous veins and tributary veins with a diameter at the saphenofemoral or saphenopopliteal junction of > or =10 mm and compare these results with a subgroup of veins with diameters of < 10 mm. METHODS: A subgroup of 17 saphenous veins with a diameter at the saphenofemoral or saphenopopliteal junction of > or =10 mm were compared with a subgroup of 98 saphenous veins with a diameter of < 10 mm at the saphenofemoral or saphenopopliteal junction for clinical efficacy and patient satisfaction at a mean 2-year follow-up. RESULTS: A mean number of 2.15 treatments utilizing an average of 8.37 mL of foam sclerosing solution (3% sodium tetradecyl sulfate) were required to close all incompetent varicose veins in the < 10-mm group versus a mean of 2.8 treatments and 13.9 mL foam (3% sodium tetradecyl sulfate) for the > or =10-mm group. A total of 27.5% of saphenous veins of < 10 mm and 37.5% of saphenous veins > or = 10 mm required a second treatment at 3 months. One-hundred percent of patients believed that their legs were successfully treated at 2-year follow-up in both groups with 94% of the < 10-mm group noting improvements in quality of life and 100% in the > or =10-mm group. CONCLUSIONS: Ultrasound-guided foam sclerotherapy is effective in treating all sizes of varicose veins with high patient satisfaction and improvement in quality of life. Duplex ultrasound findings demonstrated a small increase in failure to close the saphenofemoral or saphenopopliteal junction with increasing size of junction diameter, but this does not significantly alter the results with respect to clearance of visible varicosities and patient satisfaction with results. PMID- 15522020 TI - Body dysmorphic disorder: suggestions for detection and treatment in a surgical dermatology practice. AB - BACKGROUND: Body dysmorphic disorder is a relatively common condition in patients seeking elective surgery. Little has been written, however, in the dermatologic surgery literature about body dysmorphic disorder, where proper recognition and management of this disorder is needed during this time of increased demand for aesthetic dermatologic surgery. OBJECTIVE: The objective was to review the prevalence, demographics, clinical features, treatment approaches, and referral suggestions for patients with body dysmorphic disorder in an attempt to facilitate care of such patients in a general dermatologic surgical practice. METHODS: We reviewed the dermatologic, cosmetic surgical, and psychiatric literature regarding body dysmorphic disorder and related disorders. RESULTS: Body dysmorphic disorder is observed in 6% to 15% of dermatologic and cosmetic surgery patients and in 2% of the general population. Surgical treatment of patients with body dysmorphic disorder typically leads to no change or worsening of symptoms in the majority of patients. The use of screening questionnaires and observation for hallmark features are helpful for clinicians in managing patients with body dysmorphic disorder. Psychiatric referral is desirable, because cognitive behavioral therapy and pharmacologic intervention with selective serotonin reuptake inhibitors are often efficacious. CONCLUSIONS: Body dysmorphic disorder is often underdiagnosed and suboptimal management is common. Effective treatment consists of behavioral and pharmacologic intervention. Use of the Dufresne Body Dysmorphic Disorder Questionnaire appears to be useful as a screening tool in an outpatient setting, and awareness of clinical features of body dysmorphic disorder in the dermatologic surgical setting may spare patients significant morbidity while allowing surgical dermatologists to manage their patients and practices more effectively. PMID- 15522021 TI - Comparison of melanocytes transplantation methods for the treatment of vitiligo. AB - BACKGROUND: Surgical therapy of vitiligo is indicated when depigmented macules are localized in areas that are known to respond poorly. OBJECTIVE: The objective was to compare the results of treatment of vitiliginous macules localized in the dorsum of the hands and lower limbs by transplantation of cultured autologous melanocytes plus PUVA therapy (CMP), suction blister transplantation plus PUVA therapy (SBP), cryotherapy plus PUVA therapy (CP), and only PUVA therapy (OP). METHODS: Twenty patients qualified for the study. The patients were split into two groups of 10 patients. In the first group, the CMP procedure was performed on one limb and OP on the other. In the second group, SBP and CP were used, respectively. RESULTS: The CMP procedure was successfully performed on only 6 of 10 patients, whereas SBP was carried out on all 10 patients. No significant difference was found between the number of successful transplants in both groups of patients. A total lack of effectiveness was found in CP and OP methods. CONCLUSION: This study demonstrated the advantage of the suction blister transplantation method over the autologous cultured melanocytes transplantation method because of the difficulties in cell culture establishment in some cases. PMID- 15522022 TI - Reducing opposed multilobed flap repair, a new technique for managing medium sized low-leg defects following skin cancer surgery. AB - BACKGROUND: Following skin cancer excision, skin defects on the low leg between 10 and 35 mm in diameter can be problematic. Direct closure of the wound risks excessive wound tension and wound dehiscence. Skin grafts heal slowly and often remain unsightly. Traditional skin flaps have a limited role. OBJECTIVE: The objective was to develop a random-pattern skin flap that offers significant advantages over traditional techniques including grafting. METHODS: The reducing opposed multilobed flap involves a series of semicircular lobes extending both cephalic and caudal from the defect. The technique involves lobes most distant from the primary defect being transposed in turn closer to the defect. The technique does not result in the unnecessary excision of Burrows triangle skin. RESULTS: The reducing opposed multilobed (ROM) flap reduces skin tension concerns, lowers the risk of flap necrosis, and allows for quicker and more aesthetic healing. After 20 cases, we have yet to experience dehiscence, infection, or delayed healing. The healing wound is resistant to contraction and invariably produces an acceptable aesthetic outcome. CONCLUSION: The ROM flap repair allows the dermatologic surgeon an additional option when faced with a medium-sized lower leg defect following skin cancer excision. PMID- 15522023 TI - Basal cell carcinoma occurring in scar tissue following excision of a parotid gland pleomorphic adenoma. AB - BACKGROUND: A 49-year-old patient presented with a basal cell carcinoma growing on scar tissue left following excision of a parotid gland pleomorphic adenoma. OBJECTIVE: The objective was to determine whether the basal cell carcinoma had resulted from tumor spillage at the time of the pleomorphic adenoma excision. METHODS: Substantial evidence is presented linking the two tumors. RESULTS: The basal cell carcinoma most likely occurred as the result of tumor spillage at the time of the parotid gland pleomorphic adenoma excision. CONCLUSIONS: Cutaneous connective tissue stroma plays a significant role in determining the differentiation of basal cell carcinomas. PMID- 15522024 TI - Subcutaneous nodules on the face: acantholytic in-transit cutaneous metastasis. AB - BACKGROUND: Although skin is the biggest organ of the body, cutaneous or subcutaneous metastasis from internal malignancies or primary skin cancers is very uncommon. OBJECTIVE: In this report, we present two cases of in-transit cutaneous metastasis in acantholytic pattern from a skin tumor on the face. METHODS: Both patients presented with the subcutaneous nodule and had history of multiple squamous cell carcinomas on the face. The epidermis was intact above the nodule. Computed tomography scan was performed, and subcutaneous located cystic lesions were seen. RESULTS: Nodules were excised and reported as acantholytic squamous cell carcinoma. There was no recurrence or distant metastasis. There were no early or late complications. CONCLUSIONS: These nodules were in-transit metastasis of the previous multiple squamous cell carcinomas of the face. Although patients with squamous cell carcinomas presenting as subcutaneous nodules of the head were presented in the literature, to our knowledge, this is the first report to present in-transit metastasis of acantholytic squamous cell carcinoma. PMID- 15522025 TI - Multiple subclinical syringomatous proliferations encountered during Mohs surgery for basal cell carcinoma. AB - BACKGROUND: Examination of Mohs surgery frozen sections may lead to discovery of incidental benign neoplasms such as syringomas. OBJECTIVE: An unusual occurrence of numerous subclinical syringomatous proliferations discovered during Mohs surgery for a basal cell carcinoma that posed a diagnostic and management dilemma is reported. METHODS: Clinical records and histologic sections are examined. RESULT: Multiple syringomatous proliferations were noted around a basal cell carcinoma during Mohs surgery. A few foci were atypical appearing and focally extending into the deep dermis causing a diagnostic dilemma. All atypical syringomatous proliferations were excised. Evaluation of final Mohs margins by permanent sections and biopsies of normal-appearing facial skin showed multiple benign syringomas. CONCLUSION: Although solitary subclinical syringomas have been described in association with basal cell carcinomas, the occurrence of multiple syringomas and syringomatous proliferations has not been previously reported. Dermatologists should be aware of the existence of this phenomenon and consider permanent section evaluation to better determine nature of unusual incidental tumors identified in frozen sections during Mohs surgery. PMID- 15522026 TI - Management of squamous cell carcinoma in a patient with recessive-type epidermolysis bullosa dystrophica. AB - BACKGROUND: Surgical treatment of a recessive dystrophic epidermolysis bullosa patient is very difficult for both the surgeon and the anesthetist because of the fragility of the skin and abnormal nature of the tumor bed. OBJECTIVE: We report a case of 54-year-old Japanese recessive dystrophic epidermolysis bullosa patient with squamous cell carcinoma (SCC) of the lateral malleolus. METHODS: A tumor measuring 5.0 x 5.5 cm was surgically excised. The defect was then reconstructed by full-thickness skin grafting. To avoid airway complications, general anesthesia was administered using a face mask. Because the regional lymph nodes were swollen before surgery, the patient underwent sentinel lymph node biopsy. RESULTS: The patient remains well with no sign of recurrence or metastasis 7 months after surgery. CONCLUSION: To preserve activities of daily living, surgery should be performed for squamous cell carcinomas arising in recessive dystrophic epidermolysis bullosa patients. PMID- 15522028 TI - Re: Improvement of neck and cheek laxity with a nonablative radiofrequency device: a lifting experience. PMID- 15522030 TI - Re: Eyebrow reconstruction and subcutaneous island pedicle flaps. PMID- 15522034 TI - The need for new antibiotics. AB - Politicians and public health officials have joined specialist professionals in recognising antibiotic resistance as a threat to modern medicine. Their response has centred on minimising unnecessary antibiotic prescribing, aiming to reduce selection pressure for resistance. Despite a few hopeful trends (e.g., declining penicillin resistance among pneumococci in the UK), established resistance is proving hard to displace; moreover, new resistances continue to emerge and to proliferate at new sites. There consequently remains a strong need for new antibiotics, particularly those directed against multiresistant Gram-negative bacteria in hospitals. Already some nonfermenters of the genera Acinetobacter and Pseudomonas are resistant to all good antibiotics and many Enterobacteriaceae are resistant to all except carbapenems. There is also a growing need for new agents against community-acquired pathogens, including the agents of tuberculosis, gonorrhoea and urinary tract infections. Unless antibacterial development is re energised, there is a serious risk that a growing proportion of infections, especially in hospitals, will become effectively untreatable. PMID- 15522035 TI - Antibacterial drug discovery in the 21st century. AB - Antibacterial research over the past 50 years has been focused on meeting medical needs caused by infectious, life-threatening pathogens. In spite of the introduction of a variety of antibacterial agents in multiple unrelated drug classes, resistance continues to emerge. The pharmaceutical industry must respond to these clinical challenges by bringing forward a stream of new agents with antibacterial activity against resistant bacteria. Although the projected growth of the anti-infective area may not be as large as for some therapeutic areas, development advantages for these agents include their higher predictability for success, well-defined biomarkers, shorter clinical trials, and shorter duration of therapy leading to fewer long-term safety concerns. Anti-infectives are still attractive commercially, representing the third largest therapeutic area in terms of worldwide sales of 45 bn dollars, with growth predicted at least through 2010, particularly for the hospital-related products. Finally, companies that conduct anti-infective research demonstrate their social responsibility by developing agents to treat patients with acute and potentially fatal illnesses. PMID- 15522036 TI - Antibacterial drug discovery: is it all downhill from here? AB - There has been a marked decline in the industrial research aimed at discovering novel antibacterial agents, including new drugs that target resistant organisms. While this decline may reflect past cyclical changes that often affect resource allocation at pharmaceutical companies, this decline is occurring at a time of increasing levels of antibacterial drug resistance and meagre pipelines of new agents that are active against them. There are multiple reasons for this decline, although few are unique to antibacterial drug discovery research. These include: lack of industry productivity, increasing size of clinical trials, increased generic competition and other pressures on drug pricing, a crowded and confused marketplace and industry consolidation. And while many (if not most) large companies and biotechs have exited the field or severely curtailed their research, others have made it a point to continue their efforts, citing both the unmet medical need and a large and apparently growing market. Despite the fact that some companies have remained engaged, the view here is that the current level of industrial effort is insufficient to sustain a healthy flow of new and better agents that are needed to counter the imminent threat of bacterial drug resistance. Therefore, a clear and urgent need for finding ways to improve the level and quality of industrial research in this area is apparent. PMID- 15522037 TI - Antimicrobial drug development--the past, the present, and the future. AB - Antimicrobial resistance has been an issue since the introduction into clinical use of the first agents in the 1940s. Although the discovery and development of new classes of antimicrobials through the 1960s presented an array of treatment options, these options for some serious and life-threatening infectious diseases may now be more limited. This paper examines the history of antimicrobial development, showing how the challenges in discovering new classes of drugs have been with us for the last 40 years. The present state of antimicrobial discovery and development is shaped by these challenges as well as by the economic realities of the pharmaceutical industry. This paper also discusses some of the regulatory considerations in antimicrobial drug development, and presents some potential solutions to the challenges inherent in antimicrobial drug development, including steps taken by the US Food and Drug Administration to streamline the drug review process for antimicrobial agents while maintaining the standards necessary to protect and promote the health of the public. PMID- 15522038 TI - Antibacterial drug discovery: is small pharma the solution? AB - Although antibacterial research has declined in many larger pharmaceutical companies, small companies have begun to fill in the gap. Antibacterial discovery research is currently being conducted in at least 35 small companies. One successful approach taken by small pharma has been to continue clinical programmes that were abandoned by large companies. Issues surrounding these activities, as well as proposed changes, are outlined. PMID- 15522039 TI - Non-invasive assessment of endothelial function - relation between vasodilatory responses in skin microcirculation and brachial artery. AB - OBJECTIVE: To compare different non-invasive methods for determination of human endothelial function in peripheral circulation. DESIGN: Observational, cross sectional study in 39 healthy subjects (21 females, age 17-56 years). SETTING: Vascular research laboratory at university hospital. METHODS: Laser Doppler (LD) flowmetry was used to compare skin microvascular perfusion changes during postocclusive reactive hyperaemia with those induced by iontophoretic administration of acetylcholine (ACh), an endothelial-dependent vasodilator. LD measurements were compared with ultrasonographic measurements of postocclusive flow-mediated dilatation (FMD) in the brachial artery (n = 21). RESULTS: Local ACh induced a larger and more sustained skin perfusion increase than reactive hyperaemia after 4 min of regional arterial occlusion (P<0.001). A significant correlation was found between the magnitude of ACh-induced vasodilatation and peak reactive hyperaemia, both in absolute (r = 0.62, P<0.001) and relative terms (r = 0.58, P<0.001). A correlation was also found between brachial artery FMD and the magnitude of ACh-induced skin perfusion increase (r = 0.43, P<0.05) but not between FMD and reactive hyperaemia. CONCLUSION: Endothelial function, an early marker of cardiovascular risk, can be non-invasively assessed and graded by LD and FMD-measurements and despite inherent differences, both methods do correlate. PMID- 15522040 TI - Effect of body position on foot and ankle volume in healthy subjects. AB - Physical therapy treatments for and evaluation of lower limb swelling are usually performed in supine lying position. However, the limb is usually treated or assessed in the dependent position, i.e. sitting or standing. The purpose of this study was to determine the possible confounding effect of subject positioning on foot and ankle volume. Sixty healthy individuals were randomly positioned into motionless standing, sitting or supine lying for 30 min. Volumetric measurements of the dominant leg were taken with a foot and ankle volumeter before and after 30 min of positioning. A significant difference between the three groups for foot and ankle volume change (F(2,53) = 35.41, P<0.001) was found. A post-hoc Tukey HSD test revealed that the difference was statistically significant between sitting and standing as well as supine lying and standing (P<0.001). No statistical significance was found between sitting and supine lying (P = 0.90). Standing caused the greatest increase in foot and ankle volume followed by sitting then supine lying. The results indicate that treating or assessing a limb with the subject in a sitting position would not be significantly different from a subject who is positioned in supine lying. However, it is not known whether the height and the distance from the heart to the feet were comparable between the groups, and so it is recommended that, where possible, treatment and assessment of lower leg swelling should be undertaken with the leg in a non-dependent position and definitely not in standing. PMID- 15522041 TI - Pulmonary gas exchange during exercise in Fontan patients at a long-term follow up. AB - Desaturation during exercise in the Fontan type of circulation may be due to known right-to-left shunts in some patients, while in others there are no obvious signs of shunts. AIM: To evaluate the gas exchange and exercise capacity in patients with Fontan circulation. MATERIAL AND METHODS: Twenty patients with the Fontan type of circulation, median age at operation 7.5 years (3.0-35 years), follow-up time 12.1 years (8.3-20.4 years), were included. The intrapulmonary gas mixing (N(2)-slope) and diffusion capacity for carbon monoxide (D(LCO)) was evaluated and determination of cardiac output was performed at rest and during exercise with measurements of ventilation, oxygen uptake, respiratory rate, carbon dioxide production and heart rate. Samples for blood gases (P(a)O(2), S(a)O(2)) were obtained at rest and at the end of each work load. RESULTS: The median exercise capacity was 1.47 l min(-1). During exercise, all the patients had an alveolar-arterial oxygen partial pressure difference (P(A-a)O(2)) above 2 SD. Patients with known right-to-left shunts had lower P(a)O(2) and S(a)O(2) values and higher P(A-a)O(2) values both at rest, median 7.5 kPa, 90%, and 7.2 kPa, respectively, and during exercise, median 5.8 kPa, 77% and 10.1 kPa, compared with those with no shunts, median 11.2 kPa, 97% and 4.1, respectively, at rest and 9.7 kPa, 94%, and 6.3 during exercise (P<0.01). No correlation was found between cardiac index, the N(2)-slope or the D(LCO) at rest and the P(a)O(2) and S(a)O(2) at rest or during exercise. CONCLUSION: In patients with the Fontan type of circulation, right-to-left shunts are the major cause of desaturation during exercise. PMID- 15522042 TI - Is muscle power output a key factor in the age-related decline in physical performance? A comparison of muscle cross section, chair-rising test and jumping power. AB - Ageing compromises locomotor capacity and is associated with an increased risk of falls. Several lines of evidence indicate that both changes in muscle mass and performance are causative. Most studies, however, do not discern between effects of ageing, sedentarism and comorbidity. The present study compares the age effects in muscle cross section, force and power in physically competent self selected subjects of different age groups. A total of 169 women and 89 men between 18 and 88 years, without any disease, impairment or medication affecting the musculoskeletal system were enrolled in this study. Calf muscle cross sectional area was assessed by computed tomography. Muscle force and power were assessed by jumping mechanography. No significant correlation between muscle cross section and age was found in the men. A weak correlation in the women disappeared after correction for height. Close correlations with age, however, were found for peak force and peak power. Correction for muscle cross section or body weight further increased these correlation coefficients, particularly for peak power specific to body weight (r = 0.81 in women and r = 0.86 in men). The non-sedentarian population investigated here depicted a reduction of >50% between the age of 20 and 80 without a reduction in muscle cross section. This suggests a crucial role for muscular power in the ageing process. Possibly, the jumping mechanography as a measurement of anti-gravitational power output is a promising extension of the chair-rising test, known to be predictive for immobilization and the risk of falls. PMID- 15522043 TI - Aerobic and functional capacity in a group of healthy women: reference values and repeatability. AB - Twenty-five randomly selected, low or moderately fit and healthy women (22-44 years) rated their perceived physical capacity and performed an incremental cycle exercise test with respiratory gas analysis. The aerobic and functional capacity did not decrease with age. However, tauVo(2) increased with age. The mean value of the perceived physical capacity was 10 metabolic equivalents and that of Vo(2max) 2075 ml min(-1). The increasing anaerobic metabolism was determined at three points DX (where the rate of Vco(2) increase just exceeds the rate of Vo(2) increase), PX (where Vco(2)/Vo(2) = 1.0) and PQ (where ventilation increase disproportionately in relation to Vco(2)). The mean Vo(2) (% of Vo(2max)) at DX, PX and PQ were 1263 (63%), 1528 (73%) and 1620 (78%) ml min(-1), respectively. The mean value of deltaVo(2)/deltaW was 10.2 ml min(-1) W(-1) while that of tauVo(2) was 0.578 (age) + 15.6. Ten women performed a test and re-test on two consecutive days, and eight of these performed another re-test 4 weeks later. The repeatability was analysed and the variations were expressed as 2 SD of the differences between the tests. The variation was greater for the 4-week re-test than the day-to-day re-test regarding Vo(2max), o(2) at DX, PX and PQ, deltaVo(2)/deltaW and HR. The variation in Vo(2max), PX and deltaVo(2)/deltaW for the 4-week re-test was more than twice that of the previously reported 4-week variation for men. The considerable variation, especially for 4-week re-testing for women should be considered when evaluating the effects of exercise and rehabilitation. PMID- 15522044 TI - The validity and reliability of the Perometer in evaluating human knee volume. AB - BACKGROUND: Accurate knee swelling evaluation is important in monitoring the progress of a patient's condition and rehabilitation. Water volumetry is the current gold standard of volume assessment but can be time consuming and unsuitable for assessing patients with open wounds. No previous study has reported the potential use of an optoelectronic device in assessing knee volume. OBJECTIVES: To evaluate an optoelectronic device, the Perometer in knee volume assessment in uninjured subjects and compare it to the current gold standard of volume assessment - water displacement. MATERIALS AND METHODS: Bilateral knee measurements were measured in 20 uninjured individuals with both methods on two separate days using a one-way repeated measures design. A cylindrical object was also measured 10 times with both tools. RESULTS: The mathematically calculated volume of the cylinder was 1568 ml. The mean volume obtained by the Perometer was 1602 and 1620 ml by the volumeter. This represented an over-estimation of the true volume of 2.2% by the Perometer and 3.4% by the volumeter. A high correlation was found for repeated measures of knee volumes with both methods (r = 0.94, P<0.01). Limits of agreement (LOA) were calculated to assess the amount of agreement between the two methods and these ranged from -130 to 207 ml indicating a low degree of agreement. CONCLUSIONS: The results indicate that the Perometer is as reliable as the knee volumeter in assessing knee volume. However, the wide LOA suggest the two methods should not be used interchangeably. PMID- 15522045 TI - Reliability of three commercially available heart rate variability instruments using short-term (5-min) recordings. AB - AIM: To assess the reliability of heart rate variability (HRV) measures made by three commercially available analysers in healthy subjects. METHODS: Twenty-nine volunteers (20 males, mean age 35 +/- 13 years and nine females, mean age 29 +/- 11 years) underwent repeated HRV measures under three conditions: lying supine, standing, lying supine with controlled breathing. HRV was measured simultaneously by three instruments. Reliability was assessed statistically by calculating coefficient of variation (CV), intraclass correlation coefficient (ICC) and limits of agreement (LoA). RESULTS: A wide range of values were found for CV (1 235%) and ICC (R = 0.16-0.99) dependent on the HRV measure assessed and the position in which the measurement was made. For the most part the analysers gave similar values in each condition. The values for CV and ICC were high but within the range reported in the literature. Values for LoA were also high and showed a wide range of values. CONCLUSIONS: The similarity in measures between systems indicates that biological variation and experimental error play a major role in determining the repeatability of HRV measurements. It is therefore recommended that population-specific reliability coefficients should be published where possible and that authors should take into account the reliability of measures when making sample size calculations. PMID- 15522046 TI - Ambulatory combined pH, bile and manometric monitoring of the oesophagus in asymptomatic healthy volunteers. AB - BACKGROUND: With the development of sophisticated equipment ambulatory studies of oesophageal motor function, pH and bilirubin have gained in popularity. The aim of the study was to present reference values for combined 24 h pH, bilirubin and manometric measurements of the oesophagus. METHODS: Twenty-six (15 male) healthy volunteers without symptoms of gastro-oesophageal reflux underwent a 24-h ambulatory oesophageal combined three-channel pressure, acid and bilirubin detection. RESULTS: The subjects were studied for a median of 20 h (16-22). The median per cent time with pH < 4 for the whole measured time was 3.1 (0.8-14; 5 and 95 percentiles). Bile was detected for a median of 0.05% (0.0-8.5; 5 and 95 percentiles) of the time. Eighty-one per cent of the contractions were peristaltic, 55% of which were complete. Of these, 53% had a pressure over 30 mmHg at all three pressure points, giving an efficient peristalsis in a median of 29% (13-46; 5 and 95 percentiles) of all registered contractile patterns. No difference between the genders could be observed. CONCLUSIONS: This study provides normative data for ambulatory oesophageal manometry, pH and bilirubin studies that can be used for comparing with patients with disease. PMID- 15522047 TI - Operator dependent variability in quantitative analysis of myocardial perfusion images. AB - The purpose of this study was to evaluate variability in the quantification of myocardial perfusion images obtained by a group of experienced operators using two widely used programs. The Cedars Emory quantitative analysis program (CEqual) was used to quantify the size of perfusion defects and the Cedars-Sinai quantitative gated single-photon emission tomography program was used to quantify left ventricular function. Five patients with reversible apical defects, five with fixed apical defects and three patients with normal perfusion were selected. Eight experienced medical laboratory technologists processed the studies from raw projection data. The manual steps consisted of defining two alignment axes parallel to the long axis of the left ventricle, and for the CEqual program selecting apex and base in the short axis slices in the rest and stress studies. Wide variability between the operators in the quantification of reversibility could be seen in all three vascular territories. A range >10% was found in at least one vascular territory for nine of the 13 patients. The differences in left ventricular ejection fraction (LVEF) between operators were <5% for all 13 patients. The large variability in the quantification of reversible apical perfusion defects may influence the clinical interpretation and cause false conclusions. In contrast, inter-operator variability for the quantification of the LVEF was low. PMID- 15522048 TI - Myocardial SPECT perfusion defect size compared to infarct size by delayed gadolinium-enhanced magnetic resonance imaging in patients with acute or chronic infarction. AB - BACKGROUND: Single photon emission computed tomography (SPECT) perfusion imaging has been considered a reference method for non-invasive estimation of infarct size in man. Recently, delayed gadolinium-enhanced magnetic resonance imaging (DE MRI) has evolved as an accurate tool to quantify infarct size. Therefore, the present study was designed to compare perfusion defect size by SPECT to hyperenhanced volume by DE-MRI. METHODS: DE-MRI was performed in 30 patients. Fourteen were patients with revascularized first-time acute infarctions, eight revascularized chronic infarctions, and eight clinically referred non revascularized patients. SPECT was performed in the same patients and analysed by a commercial package. RESULTS: The hypoperfused volume by SPECT was larger than the hyperenhanced volume by DE-MRI by 8 +/- 8 ml (6% +/- 5 percentage points), 10 +/- 18 ml (6% +/- 11 percentage points), and 26 +/- 30 ml (12% +/- 10 percentage points) in the acute, chronic and clinical populations, respectively. Left ventricle wall volume was smaller by SPECT in all settings. CONCLUSION: The SPECT perfusion defect size was comparable with but generally slightly larger than the hyperenhanced volume by DE-MRI in both absolute and relative terms in patients with acute and chronic infarction. The results may be related to systematic differences between modalities but could also be influenced by biological phenomena such as wall thinning or hypoperfused but viable myocardium. PMID- 15522049 TI - Determinants of hypertension and left ventricular function in end stage renal failure: a pilot study using cardiovascular magnetic resonance imaging. AB - Cardiovascular disease is the principal cause of mortality in patients with renal failure. Left ventricular (LV) abnormalities are adverse prognostic indicators for cardiovascular outcome. The aim of this study was to use cardiac magnetic resonance scanning (CMR) to define LV functional abnormalities in haemodialysis (HD) patients and clarify the determinants of blood pressure (BP) and the effect of anaemia in this population. We studied 44 HD patients and 11 controls with CMR performed following dialysis. Forty patients and 11 controls completed the study. LV mass (P<0.001) and estimated systemic vascular resistance (SVR) (P = 0.002) were significantly higher in the dialysis group compared to controls. LV ejection fraction (P = 0.002) and SV (P = 0.043) were lower than controls. In the HD patients, BP correlated significantly with cardiac output (CO; r = 0.569, P<0.001) and end diastolic volume (EDV; r = 0.565, P<0.001) but there was no correlation between BP and SVR (r = 0.201, P = 0.594). Haemoglobin was inversely correlated with both CO (r = -0.531, P<0.001) and EDV (r = -0.493, P = 0.001) and positively with SVR (r = 0.402, P = 0.009). HD patients had a higher LV mass and lower ejection fraction than controls. The relationship of BP with CO, but not SVR, supports the theory that a major determinant of BP is intravascular volume and CO rather than vascular resistance although there was a fixed increase in SVR in this population. Improved understanding of the mechanisms underlying increased SVR and improved control of CO and intravascular volume may allow better therapeutic strategies. CMR provides insights into the pathophysiology of hypertension and LV dysfunction in HD patients. PMID- 15522050 TI - Arterial concentration of 99mTc-sestamibi at rest, during peak exercise and after dipyridamole infusion. AB - Tracers for myocardial perfusion imaging during stress should not only have high cardiac uptake but they should also have a fast blood clearance to prevent myocardial tracer uptake after the ischaemic stimulus. The present study characterize the early phase of the arterial (99m)Tc-sestamibi (MIBI) time activity curve after venous bolus injection at rest, during peak exercise and after dipyridamole infusion. We included 11 patients undergoing angioplasty for one-vessel disease (rest study) and 20 patients evaluated for the detection of haemodynamic significant coronary stenoses by (99m)Tc-sestamibi single photon emission computed tomography (SPECT) using either bicycle exercise testing (10 patients) or standard dipyridamole testing (10 patients). Arterial blood samples of 1 ml were taken from the left femoral artery (rest study) or the right radial artery (exercise and dipyridamole studies) every 5 s during the first 5 min postinjection. In the exercise and the dipyridamole studies blood sampling were extended to include blood samples every 5 min 5-30 min postinjection. Peak MIBI concentration was lower and decrease in concentration slower after tracer injection during exercise than during dipyridamole stress testing. This may cause an underestimation of perfusion defects during exercise because of MIBI uptake after the ischaemic stimulus. The implications of the study not only refer to the choice of stress modality when using MIBI. This study also underlines the importance of considering early blood clearance in addition to regional myocardial tracerkinetic aspects such as myocardial extraction fraction when new tracers are introduced. PMID- 15522051 TI - Reflections on two articles in the June 2004 edition of Dental Traumatology. PMID- 15522052 TI - Federal University of Santa Catarina follow-up management routine for traumatized primary teeth -- part 1. AB - The objective of this study was to verify if the follow-up management routine of traumatized primary teeth set up by Federal University of Santa Catarina, which performs clinical and radiographic assessments (15 and 45 days; 4, 8 and 12 months) after the oral trauma, enabled an early diagnosis of sequelae which would indicate the need for endodontic intervention, as well as the influence a type of trauma and the child's age could have in the severity of the sequelae. In this study 52 sets of records were used of patients being seen in the last 6 months, with a total of 70 teeth that were receiving follow-up treatment. Patients returned for regular visits set up by the management routine, where clinical and radiographic examinations were performed to check for sequelae, which justified endodontic intervention. Mobility (51.2%) and crown discoloration (25.6%) were the most common sequelae found in the patient's first appointment. In the follow up visits, replacement root resorption (22.5%) was the second most common sequela found, suggesting endodontic intervention. No significant association was found between severe sequelae, types of trauma and a child's age (chi(2) = 0.3, P = 0.8613). During the intervals of the follow-up visits, it was noticed that between 46 days and 8 months a higher number of sequelae were diagnosed (P < 0.05). The diagnosis of sequelae such inflammatory and replacement root resorption, which can lead to an early loss of a primary tooth, are frequent and that the interval between the follow-up visits has to be changed, suggesting the setting up of management routine 2. The study also concluded that the type of trauma and the child's age are not fundamental factors in the diagnosis of severe sequelae. PMID- 15522053 TI - Federal University of Santa Catarina endodontic treatment of traumatized primary teeth - part 2. AB - This research sought to evaluate periapical repair in 28 traumatized deciduous teeth that had suffered endodontic intervention due to the presence of internal or external inflammatory resorption or replacement root resorption. After obtaining endodontic access, work length and biomechanical preparation, the root canals were filled with calcium hydroxide and propylene glycol under the form of a dense slurry, during 12 months. Replacement of the intracanal dressing was performed when monthly radiographic examinations showed its absence. After 12 months the teeth were obturated with zinc oxide and eugenol cement. Halting of the inflammatory and replacement root resorption (64.3%; n = 28) occurred 9 months after the use of calcium hydroxide dressings, in a total of 18 successful cases. Fisher's test was applied to relate success with the type of trauma, work length time, child's age and pulpal condition. The test did not present statistical significance (P < 0.05). However, in the qualitative analysis, failure was observed in those cases (35.7%) where replacement resorption was already present at the moment of treatment (up to two-thirds) associated with severe trauma cases. The authors concluded that endodontic treatment must be initiated at an early stage, and must be coincident with the radiographic signs of resorption. Success of the treatment is directly related to the seriousness of the sequelae at the moment of the first examination or the endodontic treatment. PMID- 15522054 TI - The cost of treating children and adolescents with injuries to their permanent incisors at a dental hospital in the United Kingdom. AB - The aim was to estimate the total cost, including the direct costs (outpatient costs) and indirect costs (missed working day) of treating children and adolescents with traumatic injuries to their incisors. Factors such as the number of treatment visits and the success of outcome were also investigated. The sample was taken from patients who attended the dental trauma clinic at a London teaching hospital between 1990 and 2001. Eighty-one patients, with 111 traumatized incisors were included in this study. The mean age was 9.9 (SD = 2.33) years and the male:female ratio was 3:2. The median number of visits and median treatment duration were eight visits and 21 months, respectively. Sixty two per cent of the patients lived >5 miles and 25% lived >10 miles from the hospital; 44% of the patients had uncomplicated and 56% had complicated trauma to their incisors. Accidental falls, falls involving a second person, sport-related injuries and road accidents accounted for 30, 22, 22 and 17% of the total injuries. For uncomplicated trauma, 97% of the patients had a successful outcome but this was reduced to 58% for complicated trauma. The average total cost of treating a patient with one traumatic injury was 856 pounds. The best predictor for higher number of visits and unsuccessful outcome was complicated trauma with odd ratios of 4.5 and 24 (95% CI 1.5-13.7 and 2.9-194.2), respectively. It was concluded that the indirect cost was a considerably large proportion (39%) of the total cost. More specialists in paediatric dentistry are needed to improve access to care locally and thus reducing the indirect travelling cost. PMID- 15522055 TI - The effect on osteogenesis of type I collagen applied to experimental bone defects. AB - The purpose of the present investigation was to evaluate the effects of type I collagen sponge on the healing of bone defects. In this study, six adult male rabbits were used. After the induction of general anesthesia with intraperitoneal kethamine, the anterior surfaces of tibias of the rabbits were surgically exposed, and two holes with 4 mm in diameter were prepared on each tibia for the investigation. Only one hole in each tibia was filled with type I collagen, the other unfilled hole was used as control. During the study, radiopacity changes in the radiographs of the tibias of the rabbits were evaluated. The animals were killed on the 28th day, and histologic sections of the tibias were prepared. On the 28th day, it was histopathologically observed that collagen cavities were filled with new bone. In addition, it was determined that there was an increase in radiopacity of the defect areas from 14 to 28 days in both groups, and there were statistically a significant difference between control and collagen groups (P = 0.0001). In this study, consequently, it was determined that type I collagen sponge in the experimental cavities provides a more rapid regeneration of bone defects compared with non-filled cavities. PMID- 15522056 TI - Non-surgical treatment of sports-related temporomandibular joint disorders in basketball players. AB - In this study, 18 basketball players (11 female, seven male; age range 14-32 years) with temporomandibular joint (TMJ) problems who had a history of sport injuries related to head or jaw region were evaluated and the results of the treatment were presented. A standardized functional examination of the masticatory system was performed including measurement of maximal jaw movements, recording of joint sounds, pain on movement of the jaw as well as tenderness to palpation of the both TMJ or masticatory muscles. Patients were also evaluated periodontally. Diagnosis was made according the criteria, described by Okeson, and appropriate treatment was applied using different kind of splints. At the end of treatment only one patient continued with right TMJ reduction with slight pain. Except for that patient, none of the patients had pain after treatment. The maximum opening of the jaws and the maximal jaw movements were statistically increased after treatment. Patients with TMJ problems also showed periodontal problems, most likely due to inadequate oral hygiene because of the limited jaw movements and pain. Periodontal parameters including probing depth (PD), Plaque index (PI), and Sulcus Bleeding Index (SBI) improved after treatment. Sports related TMJ injuries may result in complex problems such as pain, TMJ sounds, limitation in maximal jaw movements and maximum opening of the mouth, difficulty in chewing. With the appropriate diagnosis this could be treated non-surgically in 6-8 months. This study also showed that the TMJ disorders may cause periodontal problems, which may affect all teeth and also the general health of the athlete. PMID- 15522057 TI - Post-traumatic use of dental implants to rehabilitate anterior maxillary teeth. AB - The treatment sequel of post-traumatic teeth for the use of dental implants in the anterior maxillary region to rehabilitate anterior maxillary missing teeth was evaluated. Files of 53 healthy patients reporting anterior dental trauma were reviewed. All patients had an anterior maxillary dental implant because of tooth loss after trauma. At initial examination, 18 patients (34%) had root canal treatment and an inflammatory lesion, 15 (28.3%) had a missing tooth on admission, 12 (22.6%) had a prior operation (i.e. root-end surgery or crown lengthening), 4 (7.5%) presented an ankylotic root, and 4 (7.5%) had a root remnant not suitable for rehabilitation, with no inflammatory periapical lesion. Treatment sequences and complications were recorded. Augmentation procedure (i.e. onlay bone graft or guided bone regeneration) was performed in 43 patients (81.1%), and 2 patients (3.8%) had orthodontic extrusion prior to tooth extraction and implantation. Implants were placed immediately in 25 patients (47.2%) and 4 (7.5%) had immediate loading at the time of implantation. Complications and postoperative incidents (fistula, inflammation, swelling hematoma, etc.) were observed in 24 patients (45.3%). There was no difference in complication and postoperative incident rates with regards to the implantation technique. Complications were found at the prosthetic phase in seven patients (13.2%; six fistula and one implant failure). When patients were divided into two groups, with and without an inflammatory lesion, a significantly lower complication and postoperative incident rate were found in the non-inflammatory group (P = 0.057). This study reaffirmed the necessity for scrupulous diagnosis of teeth and alveolar bone after a traumatic injury. Treatment is multidisciplinary, requiring surgical, orthodontic, endodontic, operative, and prosthetic compliance. A specially designed treatment plan for each patient is necessary. General rules do not apply. PMID- 15522058 TI - ITI implants and Dolder bars in the treatment of large traumatic defect of mandible: a clinical report. AB - The development of more sophisticated implant techniques to produce satisfying results improves the precise planning of both the surgical phase of the implantation and the following prosthetic rehabilitation. Ball and bar attachments are the main retainer systems for implant-bearing overdentures to achieve a successful treatment in the partial or full edentulism. In this clinical report, a 23-year-old male patient, presented with a large traumatic defect in the anterior mandible, was treated with ITI implant and ITI Dolder bar combinations. The reason to prefer this kind of treatment depends on the highest retention capacity and cleaning facilities of the system. PMID- 15522059 TI - Recommendations on haematological criteria for the diagnosis of epoetin-induced pure red cell aplasia. AB - Pure red cell aplasia (PRCA) is a rare condition characterised by an arrest in red blood cell production, which may be congenital or acquired. Recombinant human erythropoietin (epoetin) was introduced in 1989 for the treatment of anaemia of chronic kidney disease patients and has maintained an excellent therapeutic and safety record while treating hundreds of thousands of patients. A very rare, but serious adverse event associated with epoetin administration is a condition in which patients develop neutralising anti-erythropoietin antibodies and, consequently, PRCA. This condition is referred to as epoetin-induced PRCA (epo PRCA). Since it is a rare condition, many haematologists and nephrologists around the world see the condition infrequently and may be uncertain about the diagnosis. For this reason, an ad hoc international working group of expert haematologists and nephrologists met together to derive new recommendations for the haematological diagnosis of epo-PRCA. These recommendations, which represent the consensus opinions of the working group, address haematological approaches to monitor and investigate suspected epo-PRCA and should help physicians differentiate between PRCA and other bone marrow diseases, as well as, between PRCA and epo-PRCA. PMID- 15522060 TI - Effectiveness of propagermanium treatment in multiple myeloma patients. AB - Interferon (IFN) is one of several drugs effective in treating multiple myeloma (MM), and propagermanium is an IFN inducer. We report on 10 MM patients who were treated with propagermanium at doses from 10 to 40 mg. Two patients achieved complete remission (CR), two patients achieved partial remission (PR), and the condition of four patients was stable (stable disease, SD). After discontinuation of propagermanium, the status of MM progressed in two patients who were in SD and in two patients who had achieved PR. The administration of propagermanium was restarted in one patient resulting in a decrease in her paraprotein. PMID- 15522061 TI - Features of extramedullary and extraosseous multiple myeloma: a report of 19 patients from a single center. AB - Extramedullary (EM) localizations at diagnosis or during the course of multiple myeloma (MM) are rare. We conducted a large retrospective study to more accurately describe the clinical and laboratory features of this entity, and the outcome of these manifestations. The charts of 19 eligible patients out of 432 patients with MM were retrieved from the hematology department of the Institut Paoli-Calmettes Cancer Center. Median age was 61 (range: 39-79) with a female/male sex ratio of 8/11. Ten patients were found to have EM and extraosseous tumor at the time of MM diagnosis, and nine patients developed EM tumor during the course of the disease. Neither the stage of the disease, the LDH level, or the type of immunoglobulin (Ig) was found to be associated with the development of EM disease. Patients who developed EM tumor during the course of MM had a lower serum Ig and a higher monoclonal Bence-Jones proteinuria at the diagnosis of MM than patients who presented with EM tumor at diagnosis. Multiple sites were usually involved. Resistance to chemotherapy was frequent and response to thalidomide was poor. Eight out of the 19 patients responded to high-dose chemotherapy. The remaining 11 patients progressed while on therapy. With a median follow-up of 13 months (range: 2-65), six patients are alive, four patients are in partial remission and two patients in present progressive disease. In conclusion, EM tumors are a rare manifestation of MM, with a cumulative incidence of 4.6% of MM. Multiple sites are usually involved. The response to chemotherapy is very poor with a very low response rate to thalidomide. The prognosis is very poor, especially when the diagnosis of EM tumor is concurrent with the diagnosis of MM. PMID- 15522062 TI - VH gene mutation status and cellular drug resistance in chronic lymphocytic leukaemia. AB - OBJECTIVE: B-cell chronic lymphocytic leukaemia (B-CLL) can be divided into two clinical entities based on the immunoglobulin variable heavy chain (VH) gene mutation status, as cases with unmutated VH genes display a more aggressive disease with shorter survival time than mutated cases. The aim of this study was to investigate whether differences in cellular drug resistance could give an explanation for these divergent clinical courses. METHODS: The VH gene mutation status was analysed in patients with previously untreated B-CLL using VH gene family-specific PCR amplification and nucleotide sequencing. In vitro sensitivity to cytarabine, fludarabine, cladribine, doxorubicin, idarubicin, vincristine, cyclophosphamide, melphalan and prednisolone was assessed using the non clonogenic in vitro assay, fluorometric microculture cytotoxicity assay. RESULTS: The VH genes and in vitro drug resistance were successfully analysed in 46 cases, revealing that 25 (54%) cases showed unmutated and 21 (46%) cases mutated VH genes. Interestingly, the unmutated group generally tended to be more chemosensitive than the mutated group with significant differences for cytarabine and prednisolone (P < or = 0.01). CONCLUSION: The propensity of inferior drug response in mutated B-CLL may reflect a more differentiated disease than in unmutated B-CLL. We conclude that the difference in prognosis between B-CLL cases with unmutated and mutated VH genes could not be explained by difference in cellular drug resistance. PMID- 15522063 TI - Rituximab as treatment for minimal residual disease in hairy cell leukaemia. AB - Purine analogues have dramatically improved the outcome of patients affected by hairy cell leukemia (HCL), although complete eradication of disease was achieved in few cases. The purpose of this study was to evaluate the role of Rituximab in eradicating minimal residual disease (MRD) in HCL patients after a pre-treatment with 2-chloro-deoxy-adenosine (2-CdA). Ten patients received four cycles of Rituximab after administration of Cladribrine. Before starting anti-CD20 antibody, two patients were in complete remission, six in partial remission and two showed no significant response to Cladribrine. All cases resulted IgH positive. Median time from the last 2-CdA infusion was 5.7 months. Eight of 10 patients [four in partial remission (PR), two in complete remission (CR) and two unresponsive after 2-CdA] were evaluable for response. Two months after the end of anti-CD20 therapy, all evaluated patients presented a complete haematological remission. Moreover, Rituximab increased percentage of molecular remission up to 100% 1 yr after the end of treatment. Interestingly, in all cases but one, including those persistently polymerase chain reaction (PCR)-positive, semi quantitative molecular analyses showed MRD levels lower than those found before Rituximab administration. Toxicity was very mild. The present results not only confirm the therapeutic effect of Rituximab, but also show its relevance in eradicating MRD in HCL. PMID- 15522064 TI - The influence of farnesyl protein transferase inhibitor R115777 (Zarnestra) alone and in combination with purine nucleoside analogs on acute myeloid leukemia progenitors in vitro. AB - R115777 (Tipifarnib, Zarnestra)-farnesyl transferase inhibitor belongs to the new class of signal transduction inhibitors which seems to be yielding results in the treatment of patients with solid tumors. Recently it has also been considered as a drug of promise in hematologic malignancies such as acute myeloid leukemia (AML), especially in older patients, in chronic myelogenous leukemia (CML) as well as myelodysplastic syndromes (MDS). The aim of our study was to evaluate the influence of R115777 used alone or with purine nucleoside analogs (PNA): cladribine (2-CdA) and fludarabine (F-ara-A) on leukemic progenitors [colony forming unit-leukemia (CFU)-L] from AML patients. Our studies were based on the methods of semisolid leukemic CFU-L and normal granulocyte-macrophage progenitor (CFU-GM) cultures in vitro. R115777 was added to the culture alone or in combinations with PNA. We showed that R115777 used alone or together with PNA at all combinations significantly inhibited the colony growth of AML CFU-L, when compared with normal CFU-GM (P < 0.01). In addition, the drugs used in combinations of two higher concentrations in significantly higher degree inhibited CFU-L colony growth, when compared either with R115777 or with any of PNA used alone (P < 0.04). IC(50) for R115777 were 67.1 and 121.9 nm for AML CFU L and normal CFU-GM, respectively. Furthermore, in the case of AML the combination index was 0.89 and 1.16, respectively, for the combination of R115777 with 2-CdA and R115777 with F-ara-A. An additive effect on AML CFU-L cells and subadditive effect on normal CFU-GM were seen. To assess a proapoptotic effect, the drugs were added to the liquid cultures at the same concentrations as for clonogenic assays. A significant increase in the rate of apoptosis induced by combinations of drugs in comparison with single agents was observed. In conclusion, the combination of R115777 with both PNA could be more effective than the drugs used alone. However, further experimental studies on the usefulness of these combinations in the treatment of myeloid leukemia patients are warranted. PMID- 15522065 TI - Specific antibodies to mouse Sca-1- (Ly-6A/E) or Thy-1-positive haematopoietic progenitor cells induce formation of nitric oxide which inhibits subsequent colony formation. AB - Mouse bone marrow cells were exposed to specific monoclonal antibodies, so that lineage positive (Lin+) cells could be removed with magnetic beads. The Lin- cells were cultured with Sca-1 or CD90 (Thy-1) monoclonal antibodies (MoAbs) in semi-solid medium for 7 d. We found that Sca-1 MoAb suppressed colony formation (20-30%), and the effect was largely abolished by N-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase. Similar results were obtained with antibodies to CD90. The findings suggest that the unknown physiological ligands to Sca-1 and Thy-1 markers on haematopoietic progenitor cells can inhibit colony formation, with NO as a pivotal mediator. Primitive progenitors may be a primary target of this Sca-1 ligand, as the Sca-1+ cell population contains the major part of the multipotent haematopoietic stem cells. PMID- 15522066 TI - Pregnancy in essential thrombocythaemia: experience with 40 pregnancies. AB - In this study, the course of 40 pregnancies in 16 women with essential thrombocythaemia (ET) was analysed retrospectively. Of the pregnancies, 45% were complicated, 55% uncomplicated, and 62% resulted in live birth. The most common complication was spontaneous abortion during the first trimester seen in 33% of all pregnancies and comprising 72% of all complications. Two intrauterine foetal deaths occurred at weeks 22 and 28. Three pregnancies were complicated by eclampsia or pre-eclampsia. Nine of 16 women with 29 pregnancies had at least one complicated pregnancy. In seven of 16 women, all 11 pregnancies were uneventful. The non-pregnancy-related symptoms of ET or the platelet count before conception or during pregnancy did not correlate with the risk of pregnancy complications. Treatment with low-dose acetylsalicylic acid (ASA) alone during pregnancy or platelet-lowering drugs before or during pregnancy reduced the risk of complications. PMID- 15522067 TI - Late puerperal thrombohemorrhagic complications in a patient with antiphospholipid syndrome. AB - In this study, we present a case of late-puerperal onset of thrombohemorrhagic complications in a 33-yr-old woman with known antiphospholipid syndrome (APS) and heterozygosity for factor V Leiden gene mutation. Antithrombotic prophylaxis with low-molecular-weight (LMW) heparin was given since the 12th gestational week. Pregnancy and cesarean delivery were uncomplicated. Five weeks postpartum, the patient developed a severe hemorrhagic diathesis with marked thrombocytopenia accompanied by vaginal, nasal and cutaneous bleeding. A variety of autoimmune phenomena were also detected, partly at clinical presentation and partly later on, despite ongoing steroid treatment. Platelet counts recovered to normal values within a few weeks secondary to high-dose steroids and intravenous immunoglobulin administration. An ultrasound of both legs, performed because of persistent complaint of moderate calf pain, revealed bilateral deep venous thromboses (DVT). The clinical and biochemical findings were not consistent with thrombotic thrombocytopenic purpura (TTP), heparin-induced thrombocytopenia (HIT) or the 'hemolysis, elevated liver enzymes and low platelet syndrome' (HELLP). The diagnostic criteria for systemic lupus erythematosus (SLE) were not fulfilled either. The complex of thrombohemorrhagic complications and autoimmune phenomena seen in this case is unusual and not previously described in the late puerperal stage of APS-related pregnancies. PMID- 15522068 TI - Resistant pure red cell aplasia after allogeneic stem cell transplantation with major ABO mismatch treated by escalating dose donor leukocyte infusion. AB - We report a case of pure red cell aplasia (PRCA) following allogeneic stem cell transplantation (SCT) with major ABO mismatch which proved resistant to all standard treatment options such as change in immunosuppressive treatment, high dose erythropoietin (EPO) or plasma exchange. We therefore proceeded to administer five cycles of Rituximab therapy, without success. Finally, escalating doses of donor-derived leukocyte infusion (DLI) resolved the PRCA of our patient 415 d after bone-marrow transplantation (BMT) and 140 d after the first infusion of donor leukocytes. A review of the literature shows the efficacy of various treatments; the role of DLI and other treatment options are discussed. Furthermore, the underlying pathophysiological mechanisms especially with regard to the role of NK cells in alloreactivity after allogeneic SCT are explained. PMID- 15522069 TI - Alemtuzumab: effective monotherapy for simultaneous B-cell chronic lymphocytic leukaemia and Sezary syndrome. AB - The simultaneous presentation of chronic lymphocytic leukaemia (CLL) and cutaneous T-cell lymphoma (CTCL) is a very rare occurrence where optimal treatment is unknown. We present the case of a 65-yr-old man who was successfully treated with alemtuzumab monotherapy for both disorders, but at a cost of severe infectious morbidity and prolonged pancytopenia. PMID- 15522070 TI - Successful induction of molecular remission with single-agent anti-CD33 antibody (gemtuzumab ozogamicin) in chemotherapy-refractory relapse of acute myeloid leukaemia post-BMT. PMID- 15522071 TI - Smarter than the average phage. AB - The seventh cholera pandemic emerged in the poorer nations of the world towards the end of the 20th century and continues to kill thousands of people per year. The causative agent of cholera, the Gram-negative bacterium Vibrio cholera, is only pathogenic when it contains a lysogenic bacteriophage, CTXphi, that encodes the toxin responsible for inducing massive fluid loss from the human host. Site specific integration of CTXphi into chromosome I of V. cholera occurs at a site, dif, that is normally required for resolution of chromosome dimers generated by homologous recombination. An article in this issue of Molecular Microbiology reports the analysis of interactions between two host encoded recombinases, XerC and XerD, and the recombination sites involved in lysogeny. Surprisingly, recombination between the CTXphi attP site and the chromosomal dif site requires additional recombinase binding sites, downstream from the positions of strand exchange, which might play an architectural role. The positions of strand cleavage also differ significantly between the two sites, suggesting a novel recombination mechanism that implicates additional host factors in resolution of the Holliday junction intermediate. PMID- 15522072 TI - Growth versus maintenance: a trade-off dictated by RNA polymerase availability and sigma factor competition? AB - The regulatory design of higher organisms is proposed to comprise a trade-off between activities devoted to reproduction and those devoted to cellular maintenance and repair. Excessive reproduction will inevitably limit the organism's ability to resist stress whereas excessively devoted stress defence systems may increase lifespan but reduce Darwinian fitness. The trade-off is arguably a consequence of limited resources in any one organism but the nature and identity of such limiting resources are ambiguous. Analysis of global control of gene expression in Escherichia coli suggests that reproduction and maintenance activities are also at odds in bacteria and that this antagonism may be a consequence of a battle between transcription factors for limiting RNA polymerase. The outcome of this battle is regulated and depends on the nutritional status of the environment, the levels of the alarmone ppGpp, and RNA polymerase availability. This paper reviews how the concentration of RNA polymerase available for transcription initiation may vary upon shifts between growth and growth-arrest conditions and how this adjustment may differentially affect genes whose functions relate to reproduction and maintenance. PMID- 15522073 TI - Genetic screening of Hrp type III-related pathogenicity genes controlled by the HrpB transcriptional activator in Ralstonia solanacearum. AB - As in many other Gram-negative phytopathogenic bacteria, the Hrp type III secretion system is essential for the pathogenicity of Ralstonia solanacearum on host plants. The expression of most of the type III effector genes previously isolated from R. solanacearum is co-regulated with those of hrp genes by an AraC type transcriptional activator, HrpB. In order to isolate type III-related pathogenicity genes, we screened hrpB-regulated genes in R. solanacearum. Using a transposon-based system, we isolated 30 novel hpx (hrpB-dependent expression) genes outside the hrp gene cluster. Most of the hpx genes contain a PIP (plant inducible promoter) box-like motif in their putative promoter regions. Seven hpx genes encoded homologues of known type III effectors and type III-related proteins found in other animal and plant pathogens. Four encoded known enzymes, namely, glyoxalase I, Nudix hydrolase, spermidine synthase and transposase. Interestingly, six hpx genes encoded two types of leucine-rich repeat (LRR) protein. Products of the remaining genes did not show any significant homology to known proteins. We also identified two novel hrpB-regulated genes, hpaZ and hpaB, downstream of hrpY in the hrp cluster. The hpaB gene of R. solanacearum, but not hpaZ, was required for both the pathogenicity and ability to induce hypersensitive reaction on plants. We show that a hpaB null mutant still produces Hrp pili on the cell surface although it shows a typical Hrp-defective phenotype on plants. PMID- 15522074 TI - FtsK activities in Xer recombination, DNA mobilization and cell division involve overlapping and separate domains of the protein. AB - Escherichia coli FtsK is a multifunctional protein that couples cell division and chromosome segregation. Its N-terminal transmembrane domain (FtsK(N)) is essential for septum formation, whereas its C-terminal domain (FtsK(C)) is required for chromosome dimer resolution by XerCD-dif site-specific recombination. FtsK(C) is an ATP-dependent DNA translocase. In vitro and in vivo data point to a dual role for this domain in chromosome dimer resolution (i) to directly activate recombination by XerCD-dif and (ii) to bring recombination sites together and/or to clear DNA from the closing septum. FtsK(N) and FtsK(C) are separated by a long linker region (FtsK(L)) of unknown function that is highly divergent between bacterial species. Here, we analysed the in vivo effects of deletions of FtsK(L) and/or of FtsK(C), of swaps of these domains with their Haemophilus influenzae counterparts and of a point mutation that inactivates the walker A motif of FtsK(C). Phenotypic characterization of the mutants indicated a role for FtsK(L) in cell division. More importantly, even though Xer recombination activation and DNA mobilization both rely on the ATPase activity of FtsK(C), mutants were found that can perform only one or the other of these two functions, which allowed their separation in vivo for the first time. PMID- 15522075 TI - The target cell plasma membrane is a critical interface for Salmonella cell entry effector-host interplay. AB - Salmonella species trigger host membrane ruffling to force their internalization into non-phagocytic intestinal epithelial cells. This requires bacterial effector protein delivery into the target cell via a type III secretion system. Six translocated effectors manipulate cellular actin dynamics, but how their direct and indirect activities are spatially and temporally co-ordinated to promote productive cytoskeletal rearrangements remains essentially unexplored. To gain further insight into this process, we applied mechanical cell fractionation and immunofluorescence microscopy to systematically investigate the subcellular localization of epitope-tagged effectors in transiently transfected and Salmonella-infected cultured cells. Although five effectors contain no apparent membrane-targeting domains, all six localized exclusively in the target cell plasma membrane fraction and correspondingly were visualized at the cell periphery, from where they induced distinct effects on the actin cytoskeleton. Unexpectedly, no translocated effector pool was detectable in the cell cytosol. Using parallel in vitro assays, we demonstrate that the prenylated cellular GTPase Cdc42 is necessary and sufficient for membrane association of the Salmonella GTP exchange factor and GTPase-activating protein mimics SopE and SptP, which have no intrinsic lipid affinity. The data show that the host plasma membrane is a critical interface for effector-target interaction, and establish versatile systems to further dissect effector interplay. PMID- 15522076 TI - The Sm-like protein Hfq regulates polyadenylation dependent mRNA decay in Escherichia coli. AB - In Escherichia coli, the post-transcriptional addition of poly(A) tails by poly(A) polymerase I (PAP I, pcnB) plays a significant role in cellular RNA metabolism. However, many important features of this system, including its regulation and the selection of polyadenylation sites, are still poorly understood. Here we show that the inactivation of Hfq (hfq), an abundant RNA binding protein, leads to the reduction in the ability of PAP I to add poly(A) tails at the 3' termini of mRNAs containing Rho-independent transcription terminators even though PAP I protein levels remain unchanged. Those poly(A) tails that are synthesized in the absence of Hfq are shorter in length, even in the absence of polynucleotide phosphorylase (PNPase), RNase II and RNase E. In fact, the biosynthetic activity of PNPase in the hfq single mutant is enhanced and it becomes the primary polynucleotide polymerase, adding heteropolymeric tails almost exclusively to 3' truncated mRNAs. Surprisingly, both PNPase and Hfq co-purified with His-tagged PAP I under native conditions indicating a potential complex among these proteins. Immunoprecipitation experiments using PNPase- and Hfq-specific antibodies confirmed the protein-protein interactions among PAP I, PNPase and Hfq. Analysis of mRNA half-lives in hfq, deltapcnB and hfq deltapcnB mutants suggests that Hfq and PAP I function in the same mRNA decay pathway. PMID- 15522077 TI - Quorum sensing and the LysR-type transcriptional activator ToxR regulate toxoflavin biosynthesis and transport in Burkholderia glumae. AB - Burkholderia glumae BGR1 produces a broad-host range phytotoxin, called toxoflavin, which is a key pathogenicity factor in rice grain rot and wilt in many field crops. Our molecular and genetic analyses of toxoflavin-deficient mutants demonstrated that gene clusters for toxoflavin production consist of four transcriptional units. The toxoflavin biosynthesis genes were composed of five genes, toxA to toxE, as Suzuki et al. (2004) reported previously. Genes toxF to toxI, which are responsible for toxoflavin transport, were polycistronic and similar to the genes for resistance-nodulation-division (RND) efflux systems. Using Tn3-gusA reporter fusions, we found that ToxR, a LysR-type regulator, regulates both the toxABCDE and toxFGHI operons in the presence of toxoflavin as a coinducer. In addition, the expression of both operons required a transcriptional activator, ToxJ, whose expression is regulated by quorum sensing. TofI, a LuxI homologue, was responsible for the biosynthesis of both N-hexanoyl homoserine lactone and N-octanoyl homoserine lactone (C8-HSL). C8-HSL and its cognate receptor TofR, a LuxR homologue, activated toxJ expression. This is the first report that quorum sensing is involved in pathogenicity by the regulation of phytotoxin biosynthesis and its transport in plant pathogenic bacteria. PMID- 15522078 TI - Characterization of XerC- and XerD-dependent CTX phage integration in Vibrio cholerae. AB - CTXphi is a filamentous bacteriophage that encodes cholera toxin and integrates site-specifically into the larger of the two Vibrio cholerae chromosomes. The CTXphi genome lacks an integrase; instead, its integration depends on the chromosome-encoded tyrosine recombinases XerC and XerD. During integration, recombination occurs between regions of homology in CTXphi and the V. cholerae chromosome. Here, we define the elements on the phage genome (attP) and bacterial chromosome (attB) required for CTXphi integration. attB is a short sequence composed of one binding site for XerC and XerD spanning the site of recombination. Together, XerC and XerD bind to two sites within attP. While one XerC/D binding site in attP spans the core recombination region, the other site is approximately 80 bp away. Although integration occurs at the core XerC/D binding site in attP, the second site is required for CTXphi integration, suggesting it performs an architectural role in the integration reaction. In vitro cleavage reactions showed that XerC and XerD are capable of cleaving attB and attP sequences; however, additional cellular processes such as DNA replication or Holliday junction resolution by a host resolvase may contribute to integration in vivo. PMID- 15522079 TI - The Era-like GTPase TrmE conditionally activates gadE and glutamate-dependent acid resistance in Escherichia coli. AB - Escherichia coli survives pH 2 acid stress at a level rivalling Helicobacter pylori. Of the three E. coli acid resistance systems involved, the one most efficient and most studied uses isozymes of glutamate decarboxylase (GadA/GadB) to consume intracellular protons, and a glutamate:gamma-amino butyric acid (GABA) anti-porter (GadC) to expel GABA in exchange for extracellular glutamate. Because acid resistance is a critical factor in resisting stomach acidity, mechanisms that control this system are extremely important. Here we show that an Era-like, molecular switch GTPase called TrmE regulates glutamate-dependent acid resistance. Western blot analysis revealed a TrmE-dependent, glucose-induced system and a TrmE-independent, glucose-repressed pathway. Gene fusion studies indicated that the TrmE requirement for GadA/B production takes place at both the transcriptional and translational levels. TrmE controls GAD transcription by affecting the expression of GadE, the essential activator of the gadA and gadBC genes. TrmE most probably controls gadE expression indirectly by influencing the synthesis or activity of an unknown regulator that binds the gadE control region. Translational control of GAD production by TrmE appears to be more direct, affecting synthesis of the decarboxylase and the anti-porter proteins. TrmE GTPase activity was critical for both the transcriptional and translational effects. Thus, TrmE is part of an increasingly complex control network designed to integrate diverse physiological signals and forecast future exposures to extreme acid. The significance of this network extends beyond acid resistance as the target of this control, GadE, regulates numerous genes in addition to gadA/BC. PMID- 15522080 TI - Characterization of a novel Neisseria meningitidis Fur and iron-regulated operon required for protection from oxidative stress: utility of DNA microarray in the assignment of the biological role of hypothetical genes. AB - We have previously shown that in the human pathogen Neisseria meningitidis group B (MenB) more than 200 genes are regulated in response to growth with iron. Among the Fur-dependent, upregulated genes identified by microarray analysis was a putative operon constituted by three genes, annotated as NMB1436, NMB1437 and NMB1438 and encoding proteins with so far unknown function. The operon was remarkably upregulated in the presence of iron and, on the basis of gel retardation analysis, its regulation was Fur dependent. In this study, we have further characterized the role of iron and Fur in the regulation of the NMB1436 38 operon and we have mapped the promoter and the Fur binding site. We also demonstrate by mutant analysis that the NMB1436-38 operon is required for protection of MenB to hydrogen peroxide-mediated killing. By using both microarray analysis and S1 mapping, we demonstrate that the operon is not regulated by oxidative stress signals. We also show that the deletion of the NMB1436-38 operon results in an impaired capacity of MenB to survive in the blood of mice using an adult mouse model of MenB infection. Finally, we show that the NMB1436-38 deletion mutant exhibits increased susceptibility to the killing activity of polymorphonuclears (PMNs), suggesting that the 'attenuated' phenotype is mediated in part by the increased sensitivity to reactive oxygen species producing cells. This study represents one of the first examples of the use of DNA microarray to assign a biological role to hypothetical genes in bacteria. PMID- 15522081 TI - RNA-guided nucleotide modification of ribosomal and non-ribosomal RNAs in Archaea. AB - Archaea use ribonucleoprotein (RNP) machines similar to those found in the eukaryotic nucleolus to methylate ribose residues in nascent ribosomal RNA. The archaeal complex required for this 2'-O-ribose-methylation consists of the C/D box sRNA guide and three proteins, the core RNA-binding aL7a protein, the aNop56 protein and the methyltransferase aFib protein. These RNP machines were reconstituted in vitro from purified recombinant components, and shown to have methylation activity when provided with a simple target oligonucleotide, complementary to the sRNA guide sequence. To obtain a better understanding of the versatility and specificity of this reaction, the activity of reconstituted particles on more complex target substrates, including 5S RNA, tRNA(Gln) and 'double target' oligonucleotides that exhibit either direct or reverse complementarity to both the D' and D box guides, has been examined. The natural 5S and tRNA(Gln) substrates were efficiently methylated in vitro, as long as the complementarity between guide and target was about 10 base pairs in length, and lacked mismatches. Maximal activity of double guide sRNAs required that both methylation sites be present in cis on the target RNA. PMID- 15522082 TI - Identification of host-specific colonization factors of Salmonella enterica serovar Typhimurium. AB - The severity of infections caused by Salmonella enterica serovar Typhimurium varies depending on the host species. Numerous virulence genes have been identified in S. Typhimurium, largely from studies in mice, but their roles in infections of other species remain unclear. In the most comprehensive survey of its kind, through the use of signature-tagged mutagenesis of S. Typhimurium we have identified mutants that were unable to colonize calf intestines, mutants unable to colonize chick intestines and mutants unable to colonize both species. The type three secretion systems encoded on Salmonella pathogenicity islands (SPIs) 1 and 2 were required for efficient colonization of cattle. However, disruption of these secretion systems only caused a minor defect in S. Typhimurium colonization of chicks. Transposon insertions in SPI-4 compromised S. Typhimurium colonization of cattle, but not chicks. This is the first data confirming a role for SPI-4 in pathogenesis. We have also been able to ascribe a role in colonization for cell surface polysaccharides, cell envelope proteins, and many 'housekeeping' genes and genes of unknown function. We conclude that S. Typhimurium uses different strategies to colonize calves and chicks. This has major implications for vaccine design. PMID- 15522083 TI - In vivo effect of inactivation of ribosome recycling factor - fate of ribosomes after unscheduled translation downstream of open reading frame. AB - The post-termination ribosomal complex is disassembled by ribosome recycling factor (RRF) and elongation factor G. Without RRF, the ribosome is not released from mRNA at the termination codon and reinitiates translation downstream. This is called unscheduled translation. Here, we show that at the non-permissive temperature of a temperature-sensitive RRF strain, RRF is lost quickly, and some ribosomes reach the 3' end of mRNA. However, instead of accumulating at the 3' end of mRNA, ribosomes are released as monosomes. Some ribosomes are transferred to transfer-messenger RNA from the 3' end of mRNA. The monosomes thus produced are able to translate synthetic homopolymer but not natural mRNA with leader and canonical initiation signal. The pellet containing ribosomes appears to be responsible for rapid but reversible inhibition of most but not all of protein synthesis in vivo closely followed by decrease of cellular RNA and DNA synthesis. PMID- 15522084 TI - Alternative oxygen-dependent and oxygen-independent ribonucleotide reductases in Streptomyces: cross-regulation and physiological role in response to oxygen limitation. AB - Ribonucleotide reductases (RNRs) catalyse the conversion of ribonucleotides to deoxyribonucleotides and are essential for de novo DNA synthesis and repair. Streptomyces spp. contain genes coding for two RNRs. We show here that the Streptomyces coelicolor M145 nrdAB genes encoding an oxygen-dependent class I RNR are co-transcribed with nrdS, which encodes an AraC-like regulatory protein. Likewise, the class II oxygen-independent RNR nrdJ gene forms an operon with a likely regulatory gene, nrdR, which encodes a protein possessing an ATP-cone domain like those present in the allosteric activity site of many class Ia RNRs. Deletions in nrdB and nrdJ had no discernible effect on growth individually, but abolition of both RNR systems, using hydroxyurea to inactivate the class Ia RNR (NrdAB) in the nrdJ deletion mutant, was lethal, establishing that S. coelicolor possesses just two functional RNR systems. The class II RNR (NrdJ) may function to provide a pool of deoxyribonucleotide precursors for DNA repair during oxygen limitation and/or for immediate growth after restoration of oxygen, as the nrdJ mutant was slower in growth recovery than the nrdB mutant or the parent strain. The class Ia and class II RNR genes show complex regulation. The nrdRJ genes were transcribed some five- to sixfold higher than the nrdABS genes in vegetative growth, but when nrdJ was deleted, nrdABS transcription was upregulated by 13 fold. In a reciprocal experiment, deletion of nrdB had little effect on nrdRJ transcription. Deletion of nrdR caused a dramatic increase in transcription of nrdJ and to a less extent nrdABS, whereas disruption of cobN, a gene required for synthesis of coenzyme B12 a cofactor for the class II RNR, caused similar upregulation of transcription of nrdRJ and nrdABS. In contrast, deletion of nrdS had no detectable effect on transcription of either set of RNR genes. These results establish the existence of control mechanisms that sense and regulate overall RNR gene expression. PMID- 15522085 TI - A second-site suppressor of a folding defect functions via interactions with a chaperone network to improve folding and assembly in vivo. AB - Single amino acid substitutions in a protein can cause misfolding and aggregation to occur. Protein misfolding can be rescued by second-site amino acid substitutions called suppressor substitutions (su), commonly through stabilizing the native state of the protein or by increasing the rate of folding. Here we report evidence that su substitutions that rescue bacteriophage P22 temperature sensitive-folding (tsf) coat protein variants function in a novel way. The ability of tsf:su coat proteins to fold and assemble under a variety of cellular conditions was determined by monitoring levels of phage production. The tsf:su coat proteins were found to more effectively utilize P22 scaffolding protein, an assembly chaperone, as compared with their tsf parents. Phage-infected cells were radioactively labelled to quantify the associations between coat protein variants and folding and assembly chaperones. Phage carrying the tsf:su coat proteins induced more GroEL and GroES, and increased formation of protein:chaperone complexes as compared with their tsf parents. We propose that the su substitutions result in coat proteins that are more assembly competent in vivo because of a chaperone-driven kinetic partitioning between aggregation-prone intermediates and the final assembled state. Through more proficient use of this chaperone network, the su substitutions exhibit a novel means of suppression of a folding defect. PMID- 15522086 TI - In vivo selection for Leishmania donovani miniexon genes that increase virulence in Leishmania major. AB - Different species of Leishmania are responsible for the diverse pathologies associated with leishmaniasis including Leishmania donovani which results in fatal visceral infection and Leishmania major which causes non-fatal cutaneous infection. In an attempt to identify genotypic differences between these related Old World Leishmania species which contribute to their distinct phenotypic characteristics, we have introduced a L. donovani cosmid library into L. major to select for L. donovani sequences which may increase L. major virulence in BALB/c mice. Through this approach, we have identified a region of the L. donovani genome which increased virulence in both visceral and cutaneous sites and was divergent from the corresponding region of the L. major genome. When these L. donovani sequences were reintroduced into L. major, they enhanced the overall virulence of L. major, increasing its ability to survive in both visceral and cutaneous sites. The region responsible for increased infection levels was determined to be the miniexon gene array derived from chromosome 36 of L. donovani. Pulse field electrophoresis revealed that L. donovani contained miniexon gene sequences in several chromosome locations as opposed to L. major which contains miniexon gene sequences only in chromosome 2. Because of the requirement for miniexon-derived transcripts in maturation of pre-mRNAs in trypanosomatids, this observation suggests that the increased expression of miniexon genes is associated with increased virulence. As the genome sequence for Leishmania becomes available, the in vivo selection procedure described within will be useful to identify additional species-specific sequences responsible for different pathogenic phenotypes associated with Leishmania infection. PMID- 15522087 TI - Enolase in the RNA degradosome plays a crucial role in the rapid decay of glucose transporter mRNA in the response to phosphosugar stress in Escherichia coli. AB - The ptsG mRNA encoding the major glucose transporter is rapidly degraded in an RNase E-dependent manner in response to the accumulation of glucose 6-P or fructose 6-P when the glycolytic pathway is blocked at its early steps in Escherichia coli. RNase E, a major endonuclease, is associated with polynucleotide phosphorylase (PNPase), RhlB helicase and a glycolytic enzyme, enolase, which bind to its C-terminal scaffold region to form a multienzyme complex called the RNA degradosome. The role of enolase within the RNase E-based degradosome in RNA decay has been totally mysterious. In this article, we demonstrate that the removal of the scaffold region of RNase E suppresses the rapid degradation of ptsG mRNA in response to the metabolic stress without affecting the expression of ptsG mRNA under normal conditions. We also demonstrate that the depletion of enolase but not the disruption of pnp or rhlB eliminates the rapid degradation of ptsG mRNA. Taken together, we conclude that enolase within the degradosome plays a crucial role in the regulation of ptsG mRNA stability in response to a metabolic stress. This is the first instance in which a physiological role for enolase in the RNA degradosome has been demonstrated. In addition, we show that PNPase and RhlB within the degradosome cooperate to eliminate short degradation intermediates of ptsG mRNA. PMID- 15522088 TI - Involvement of a novel transcriptional activator and small RNA in post transcriptional regulation of the glucose phosphoenolpyruvate phosphotransferase system. AB - RyaA is a small non-coding RNA in Escherichia coli that was identified by its ability to bind tightly to the RNA chaperone Hfq. This study reports the role of RyaA in mediating the cellular response to glucose-specific phosphoenolypyruvate phosphotransferase system (PTS)-dependent phosphosugar stress. Aiba and co workers have shown that a block in the metabolism of glucose 6-phosphate causes transient growth inhibition and post-transcriptional regulation of ptsG, encoding the glucose-specific PTS transporter. We found that RyaA synthesis was induced by a non-metabolizable glucose phosphate analogue and was necessary for relief of the toxicity of glucose phosphate stress. Expression of RyaA was sufficient to cause a rapid loss of ptsG mRNA, probably reflecting degradation of the message mediated by RyaA:ptsG pairing. The ryaA gene was renamed sgrS, for sugar transport-related sRNA. Expression of sgrS is regulated by a novel transcriptional activator, SgrR (formerly YabN), which has a putative DNA-binding domain and a solute-binding domain similar to those found in certain transport proteins. Our results suggest that under conditions of glucose phosphate accumulation, SgrR activates SgrS synthesis, causing degradation of ptsG mRNA. Decreased ptsG mRNA results in decreased production of glucose transport machinery, thus limiting further accumulation of glucose phosphate. PMID- 15522089 TI - A novel sensor kinase-response regulator hybrid regulates type III secretion and is required for virulence in Pseudomonas aeruginosa. AB - The type III secretion system (TTSS) of Pseudomonas aeruginosa is induced by contact with eukaryotic cells and by growth in low-calcium media. We have identified a protein, RtsM, that is necessary for expression of the TTSS genes in P. aeruginosa. RtsM possesses both histidine kinase and response regulator domains common to two-component signalling proteins, as well as a large predicted periplasmic domain and seven transmembrane domains. Deletion of rtsM resulted in a defect in production and secretion of the type III effectors. Northern blot analysis revealed that mRNAs encoding the effectors ExoT and ExoU are absent in the DeltartsM strain under TTSS-inducing conditions. Using transcriptional fusions, we demonstrated that RtsM is required for transcription of the operons encoding the TTSS effectors and apparatus in response to calcium limitation or to host cell contact. The operon encoding the TTSS regulator ExsA does not respond to calcium limitation, but the basal transcription rate of this operon was lower in deltartsM than in the wild-type parent, PA103. The defect in TTSS effector production and secretion of deltartsM could be complemented by overexpressing ExsA or Vfr, two transcriptional activators involved in TTSS regulation. DeltartsM was markedly less virulent than PA103 in a murine model of acute pneumonia, demonstrating that RtsM is required in vivo. We propose that RtsM is a sensor protein at the start of a signalling cascade that induces expression of the TTSS in response to environmental signals. PMID- 15522090 TI - Functional dependence on calcineurin by variants of the Saccharomyces cerevisiae vacuolar Ca2+/H+ exchanger Vcx1p. AB - The Ca(2+)-dependent protein phosphatase calcineurin is an important regulator of ion transporters from many organisms, including the Saccharomyces cerevisiae vacuolar Ca(2+)/H(+) exchanger Vcx1p. In yeast and plants, cation/H(+) exchangers are important in shaping cytosolic Ca(2+) levels involved in signal transduction and providing tolerance to potentially toxic concentrations of cations such as Ca(2+), Mn(2+) and Cd(2+). Previous genetic evidence suggested Vcx1p is negatively regulated by calcineurin. By utilizing direct transport measurements into vacuolar membrane vesicles, we demonstrate that Vcx1p is a low-affinity Ca(2+) transporter and may also function in Cd(2+) transport, but cannot transport Mn(2+). Furthermore, direct Ca(2+) transport by Vcx1p is calcineurin sensitive. Using a yeast growth assay, a mutant allele of VCX1 (VCX1 S204A/L208P), termed VCX1-M1, was previously found to confer strong Mn(2+) tolerance. Here we demonstrate that this Mn(2+) tolerance is independent of the Ca(2+)/Mn(2+)-ATPase Pmr1p and results from Mn(2+)-specific vacuolar transport activity of Vcx1-M1p. This Mn(2+) transport by Vcx1-M1p is calcineurin dependent, although the localization of Vcx1-M1p to the vacuole appears to be calcineurin independent. Additionally, we demonstrate that mutation of L208P alone is enough to confer calcineurin-dependent Mn(2+) tolerance. This study demonstrates that calcineurin can positively regulate the transport of cations by VCX1-M1p. PMID- 15522091 TI - Direct and indirect transcriptional activation of virulence genes by an AraC-like protein, PerA from enteropathogenic Escherichia coli. AB - The plasmid-encoded Per regulatory locus of enteropathogenic Escherichia coli (EPEC) is generally considered to consist of three genes, perA, perB and perC. PerA, a member of the AraC-like family of transcriptional regulators, is known to be an activator of its own promoter (autoactivation) as well as of the plasmid located bfp operon encoding bundle-forming pili, but its role in activation of the chromosomal locus of enterocyte effacement (LEE) pathogenicity island, which confers the property of intimate adherence on EPEC, requires clarification. Here, we show that PerA is also required for activation of the master regulatory LEE operon, LEE1, but that this activation is indirect, being achieved via autoactivation of the per promoter which ensures sufficient production of the PerC protein to activate LEE1. In contrast, PerA-dependent activation of the per and bfp promoters is direct and does not require the other Per proteins, but is modulated by the nucleoid-associated protein H-NS. The closely related VirF regulator from Shigella flexneri cannot substitute for PerA to activate these promoters, despite being able to bind their upstream regions in vitro. PerA can bind the per and bfp promoter fragments to form multiple complexes, while VirF forms only a single complex. Site-directed mutagenesis of the PerA protein suggests that, like VirF, it may use both of its carboxy-terminal helix-turn helix motifs for DNA interaction, and may also make direct contacts with RNA polymerase. In addition, we have isolated mutations in the poorly characterized amino-terminal domain of PerA which affect its ability to activate gene expression. PMID- 15522093 TI - Intracellular membrane transport systems in Trypanosoma brucei. AB - Trypanosomes belong to the order kinetoplastida, an early diverging group of organisms in the eukaryotic lineage. The principal reasons for interest in these organisms are twofold; they provide a superb distant triangulation point from which to assess global features of eukaryotic biology and, more importantly, they are representative of a number of pathogenic parasitic protozoa with a huge public health impact --Trypanosoma brucei, T. cruzi and Leishmania spp. Recent advances in the study of intracellular transport in T. brucei have been considerable, and a fuller picture of the complexity, function and role that the endomembrane system plays in trypanosomes is finally emerging. PMID- 15522094 TI - Location, location, location: trafficking and function of secreted proteases of Toxoplasma and Plasmodium. AB - The Apicomplexan parasites Toxoplasma gondii and Plasmodium species are obligate intracellular parasites that rely upon unique secretory organelles for invasion and other specialized functions. Data is emerging that proteases are critical for the biogenesis of micronemes and rhoptries, regulated secretory organelles reminiscent of dense core granules and secretory lysosomes of higher eukaryotes. Proteases targeted to the Plasmodium food vacuole, a unique organelle dedicated to hemoglobin degradation, are also critical to parasite survival. Thus study of the targeting and function of the proteases of the Apicomplexa provides a fascinating model system to understand regulated secretion and secretory organelle biogenesis. PMID- 15522095 TI - Functional and physical interactions between autonomously replicating sequence binding factor 1 and the nuclear transport machinery. AB - Autonomously replicating sequence-binding factor 1 (Abf1p) is a site-specific DNA binding protein in Saccharomyces cerevisiae that functions to regulate multiple nuclear events including DNA replication, transcriptional activation, and gene silencing. Previous work indicates that the multiple functions of Abf1p are conferred by the carboxy-terminus of the protein, which can be further dissected into two important clusters of amino acid residues (CS1 and CS2). Here we present genetic and cell biological evidence for a critical role of CS1 in proper nuclear localization of Abf1p. Mutations in CS1 cause severe defects in cell growth, nuclear translocation, and Abf1p-mediated gene regulation, which can be rescued by a heterologous nuclear localization sequence (NLS). In addition, the CS1 domain can mediate the import of a CS1-GFP fusion protein. Importantly, the CS1 mediated nuclear import depends on the Ran guanine nucleotide exchange factor Prp20p. Interestingly, a single amino acid change in CS1 (K625I) also causes the protein to be exported out of the nucleus via the Crm1p-dependent pathway. The temperature-sensitive growth phenotype of this particular mutant can be overcome by overexpression of Kap121p/Pse1p, a well-established nuclear transport receptor. Biochemical studies indicate that Pse1p binds to a region of Abf1p upstream of CS1 in a RanGTP-sensitive manner, suggesting that Abf1p has a second distinct NLS and can be imported into the nucleus by several overlapping pathways. We propose that the link between Abf1p and the nuclear transport machinery may also be important for partitioning multiple Abf1p-mediated nuclear processes. PMID- 15522096 TI - 3-D Structure of multilaminar lysosomes in antigen presenting cells reveals trapping of MHC II on the internal membranes. AB - In late endosomes and lysosomes of antigen presenting cells major histocompatibility complex class II (MHC II) molecules bind peptides from degraded internalized pathogens. These compartments are called MHC class II compartments (MIICs), and from here peptide-loaded MHC II is transported to the cell surface for presentation to helper T-lymphocytes to generate an immune response. Recent studies from our group in mouse dendritic cells indicate that the MHC class II on internal vesicles of multivesicular late endosomes or multivesicular bodies is the main source of MHC II at the plasma membrane. We showed that dendritic cell activation triggers a back fusion mechanism whereby MHC II from the inner membranes is delivered to the multivesicular bodies' outer membrane. Another type of MIIC in B-lymphocytes and dendritic cells is more related to lysosomes and often appears as a multilaminar organelle with abundant MHC II-enriched internal membrane sheets. These multilaminar lysosomes have a functioning peptide-loading machinery, but to date it is not clear whether peptide-loaded MHC II molecules from the internal membranes can make their way to the cell surface and contribute to T cell activation. To obtain detailed information on the membrane organization of multilaminar lysosomes and investigate possible escape routes from the lumen of this organelle, we performed electron tomography on cryo-immobilized B-lymphocytes and dendritic cells. Our high-resolution 3-D reconstructions of multilaminar lysosomes indicate that their membranes are organized in such a way that MHC class II may be trapped on the inner membranes, without the possibility to escape to the cell surface. PMID- 15522097 TI - Differential use of two AP-3-mediated pathways by lysosomal membrane proteins. AB - The adaptor protein complex AP-3 is involved in the sorting of lysosomal membrane proteins to late endosomes/lysosomes. It is unclear whether AP-3-containing vesicles form at the trans-Golgi network (TGN) or early endosomes. We have compared the trafficking routes of endolyn/CD164 and 'typical' lysosomal membrane glycoproteins (lgp120/lamp-1 and CD63/lamp-3) containing cytosolic YXXPhi targeting motifs preceded by asparagine and glycine, respectively. Endolyn, which has a NYHTL-motif, is concentrated in lysosomes, but also occurs in endosomes and at the cell surface. We observed predominant interaction of the NYHTL-motif with the mu-subunits of AP-3 in the yeast two-hybrid system. Endolyn was mislocalized to the cell surface in AP-3-deficient pearl cells, confirming a major role of AP 3 in endolyn traffic. However, lysosomal delivery of endolyn (or a NYHTL reporter), but not GYXXPhi-containing proteins, was practically abolished when AP 2-mediated endocytosis or traffic from early to late endosomes was inhibited in NRK and 3T3 cells. This indicates that endolyn is mostly transported along the indirect lysosomal pathway (via the cell surface), rather than directly from the TGN to late endosomes/lysosomes. Our results suggest that AP-3 mediates lysosomal sorting of some membrane proteins in early endosomes in addition to sorting of proteins with intrinsically strong AP-3-interacting lysosomal targeting motifs at the TGN. PMID- 15522098 TI - The function of the endocytic scaffold protein Pan1p depends on multiple domains. AB - Pan1p is an essential protein of the yeast Saccharomyces cerevisiae that is required for the internalization step of endocytosis and organization of the actin cytoskeleton. Pan1p, which binds several other endocytic proteins, is composed of multiple protein-protein interaction domains including two Eps15 Homology (EH) domains, a coiled-coil domain, an acidic Arp2/3-activating region, and a proline-rich domain. In this study, we have induced high-level expression of various domains of Pan1p in wild-type cells to assess the dominant consequences on viability, endocytosis, and actin organization. We found that the most severe phenotypes, with blocked endocytosis and aggregated actin, required expression of nearly full length Pan1p, and also required the endocytic regulatory protein kinase Prk1p. The central coiled-coil domain was the smallest fragment whose overexpression caused any dominant effects; these effects were more pronounced by inclusion of the second EH domain. Co-overexpressing nonoverlapping amino- and carboxy-terminal fragments did not mimic the effects of the intact protein, whereas fragments that overlapped within the coiled-coil region could. Yeast two-hybrid and in vivo coimmunoprecipitation analyses suggest that Pan1 may form dimers or higher order oligomers. Collectively, our data support a view of Pan1p as a dimeric/oligomeric scaffold whose functions require both the amino- and carboxy-termini, linked by the central region. PMID- 15522099 TI - Lipid raft-dependent targeting of the influenza A virus nucleoprotein to the apical plasma membrane. AB - Influenza virus acquires a lipid raft-containing envelope by budding from the apical surface of epithelial cells. Polarised budding involves specific sorting of the viral membrane proteins, but little is known about trafficking of the internal virion components. We show that during the later stages of virus infection, influenza nucleoprotein (NP) and polymerase (the protein components of genomic ribonucleoproteins) localised to apical but not lateral or basolateral membranes, even in cell types where haemagglutinin was found on all external membranes. Other cytosolic components of the virion either distributed throughout the cytoplasm (NEP/NS2) or did not localise solely to the apical plasma membrane in all cell types (M1). NP localised specifically to the apical surface even when expressed alone, indicating intrinsic targeting. A similar proportion of NP associated with membrane fractions in flotation assays from virus-infected and plasmid-transfected cells. Detergent-resistant flotation at 4 degrees C suggested that these membranes were lipid raft microdomains. Confirming this, cholesterol depletion rendered NP detergent-soluble and furthermore, resulted in its partial redistribution throughout the cell. We conclude that NP is independently targeted to the apical plasma membrane through a mechanism involving lipid rafts and propose that this helps determine the polarity of influenza virus budding. PMID- 15522100 TI - Disease-causing V(2) vasopressin receptors are retained in different compartments of the early secretory pathway. AB - The G protein-coupled V(2) vasopressin receptor is crucially involved in water reabsorption in the renal collecting duct. Mutations in the human V(2) vasopressin receptor gene cause nephrogenic diabetes insipidus. Many of the disease-causing mutants are retained intracellularly by the quality control system of the early secretory pathway. It was previously thought that quality control system is restricted to the endoplasmic reticulum (ER). Here, we have examined the retention mechanisms of eight V(2) vasopressin receptor mutants. We show that mutants L62P, DeltaL62-R64 and S167L are trapped exclusively in the ER. In contrast, mutants R143P, Y205C, InsQ292, V226E and R337X reach the ER/Golgi intermediate compartment (ERGIC) and are rerouted to the ER. The ability of the mutant receptors to reach the ERGIC is independent of their expression levels. Instead, it is determined by their folding state. Mutant receptors in the ERGIC may be sorted into retrograde transport vesicles by an interaction of an RXR motif in the third intracellular loop with the coatomer complex I. Our data show that disease-causing mutants of a particular membrane protein may be retained in different compartments of the early secretory pathway and that the folding states of the proteins determine their retention mechanism. PMID- 15522101 TI - Protein kinase A affects trafficking of the vesicular monoamine transporters in PC12 cells. AB - Previous studies have shown that the vesicular monoamine transporter 2 (VMAT2) is localized to both large dense core vesicles and synaptic vesicles in vivo. However, when exogenously expressed in PC12 cells, VMAT2 localizes only to large dense core vesicles. This distribution is similar to that of the endogenous vesicular monoamine transporter 1 (VMAT1) in PC12 cells. When VMAT2 was expressed in a protein kinase A (PKA)-deficient PC12 cell line it localized to synaptic like microvesicles. Expression of recombinant VMAT1 in the same cell line showed a heterogeneous distribution to both large dense core vesicles and synaptic-like microvesicles. Coexpression of the PKA catalytic subunit partially restored trafficking of both VMAT2 and VMAT1 to large dense core vesicles; treatment of wild-type PC12 cells with the PKA inhibitor H89 increased VMAT2 on synaptic-like microvesicles. The VMAT1 and VMAT2 in large dense core vesicles exhibit a larger molecular size than those located on synaptic-like microvesicles. This difference is due to differential N-linked glycosylation. In vitro phosphorylation experiments show that PKA does not phosphorylate VMAT2. A chimera containing the VMAT2 cytoplasmic C-terminus fused to vesicular acetylcholine transporter (VAChT) shows mislocalization to synaptic-like microvesicles and VAChT-like glycosylation in the PKA-deficient cell line. However, coexpression with PKA changes the chimera's trafficking to large dense core vesicles and increases the molecular size. These results suggest that protein kinase A affects the formation and/or composition of VMAT trafficking complexes. PMID- 15522102 TI - Ncr1p, the yeast ortholog of mammalian Niemann Pick C1 protein, is dispensable for endocytic transport. AB - The Niemann Pick C1 protein localizes to late endosomes and plays a key role in the intracellular transport of cholesterol in mammalian cells. Cholesterol and other lipids accumulate in a lysosomal or late endosomal compartment in cells lacking normal NPC1 function. Other than accumulation of lipids, defects in lysosomal retroendocytosis, sorting of a multifunctional receptor and endosomal movement have also been detected in NPC1 mutant cells. Ncr1p is an ortholog of NPC1 in the budding yeast Saccharomyces cerevisiae. In this study, we show that Ncr1p is a vacuolar membrane protein that transits through the biosynthetic vacuolar protein sorting pathway, and that it can be solubilized by Triton X-100 at 4 degrees C. Using well-established assays, we demonstrate that the absence of Ncr1p had no effect on fluid phase and receptor- mediated endocytosis, biosynthetic delivery to the vacuole, retrograde transport from endosome to Golgi and ubiquitin- and nonubiquitin-dependent multivesicular body sorting. We conclude that Ncr1p does not have an essential role in known endocytic transport pathways in yeast. PMID- 15522105 TI - Prevention of Legionnaires' disease in transplant recipients: recommendations for a standardized approach. PMID- 15522106 TI - Sequential cytomegalovirus antigenemia monitoring in kidney transplant patients treated with antilymphocyte antibodies. AB - BACKGROUND: Antilymphocyte antibodies (ALA) use is related to disseminated cytomegalovirus (CMV) disease after kidney transplantation. Strict surveillance of CMV infection, preemptive antiviral treatment or concomitant ganciclovir and ALA use are proposed as an attempt to prevent related clinical complications. Our objective was to describe the pattern of CMV infection, based on sequential antigenemia detection, after ALA treatment. PATIENTS AND METHODS: Thirty renal transplant patients were prospectively screened for CMV infection after ALA treatment. CMV antigenemia (pp65 antigen detection) was monitored twice a week in the first month and weekly until 60 days after the beginning of ALA therapy. Any positive value of antigenemia was considered CMV infection. RESULTS: Twenty-eight (93.3%) patients were CMV positive (IgG) before transplantation. The mean duration of ALA treatment was 12.1+/-2.4 days. Positive antigenemia was detected in 24 (80%) patients, a mean of 52.5+/-15 days after transplant and 44.7+/-14 days after the beginning of ALA treatment. The median antigenemia count was 7 positive cells/300,000 neutrophils (range: 1-227). Antigenemia preceded clinical symptoms by 5.8 days (0-28 days). Eighteen (75%) of 24 positive patients received ganciclovir treatment: 8 patients (26.7%) for viral syndrome, 2 patients (33.3%) for invasive disease, and 8 patients (26.7%) as part of preemptive therapy, asymptomatic with high antigenemia values. Six pp65-positive patients with low counts were followed up until a negative result and remained asymptomatic without any specific treatment. CONCLUSION: CMV infection was frequent after ALA treatment in this group and generally occurred late after completion of treatment. Antigenemia was a reliable tool to guide preemptive treatment in these patients, and such strategy is an alternative option compared to the prophylactic use of ganciclovir with ALA treatment. PMID- 15522107 TI - Antibodies against human herpesvirus 8 in South African renal transplant recipients and blood donors. AB - OBJECTIVE: To examine the seroprevalence of human herpesvirus 8 (HHV-8) antibodies in a selected group of renal transplant recipients and blood donors, in relation to various socio-demographic variables. SUBJECTS: Anonymised serum samples were obtained from 306 black donors attending blood donations at the SA National Blood Transfusion Service, Johannesburg, and 430 recipients of renal transplants at Johannesburg General Hospital, from 1998 to 1999. METHODOLOGY: Serum samples were tested by indirect immunofluorescence assay to detect IgG antibodies against HHV-8. Crude and adjusted seroprevalence, and odds ratios were calculated to examine the relationship between antibodies to HHV-8 and age, sex, population, and risk group. RESULTS: Adjusted HHV-8 seroprevalence at 'medium to high' antibody titres (at least 1:51,200) increased with age, from 2% (age 15-24; 95% CI=0.7-5.2) to 10% (age 45+; 95% CI=6.3-15.2). Whites had the lowest % adjusted seroprevalence (1.8; 95% CI:0.8-4.2), followed by blacks (7.4; 95% CI=5.1-10.8), Asians/Indians (9.5; 95% CI=3.6-22.9), and individuals of mixed race (12.5; 95% CI=4.5-30.2). Seroprevalence was not related to gender, or to occupation of blood donors. HHV-8 seroprevalence did not differ significantly between first-time blood donors, repeat donors, and transplant recipients (P=0.49). Transplant recipients had a greater proportion of persons with high HHV 8 antibody intensity; however, this difference was statistically not significant (P=0.08). CONCLUSIONS: Blood donors and renal transplant patients had relatively high HHV-8 antibody titres. Given the strong association between HHV-8 seropostivity and the risk of developing Kaposi's sarcoma, it may be prudent in settings where HHV-8 seroprevalence is high to screen for HHV-8 in donated blood used for these immunocompromised individuals. PMID- 15522108 TI - Coccidioidal pneumonia, arthritis, and soft-tissue infection after kidney transplantation. AB - A 60-year-old male kidney transplant recipient presented with pneumonia 4 months after transplantation. Coccidioidomycosis was suspected and empirically treated with fluconazole. He was maintained on suppressive fluconazole without problems. Three weeks after discontinuing secondary prophylaxis, the patient experienced coccidioidal arthritis and an infection of the soft tissue of the hand that required debridement. Transplant recipients may have quiescent disseminated coccidioidomycosis that is reactivated by immunosuppression after withdrawal of suppressive antifungal therapy. PMID- 15522109 TI - Cavitary Legionella pneumonia in a liver transplant recipient. AB - This report describes the clinical course of a liver transplant recipient in whom cavitary pneumonia developed due to Legionella pneumophila. We review the experience with cavitary pulmonary processes caused by Legionella species in liver allograft recipients and describe the diagnostic microbiology of this organism. The clinical course of this patient demonstrates the importance of considering legionellosis in the differential diagnosis of lung abscesses after liver transplantation and the diagnostic difficulties encountered with this bacterium. PMID- 15522110 TI - Cytomegalovirus-associated bleeding diathesis in renal transplant recipients. AB - Cytomegalovirus (CMV) infection is reported to be capable of modifying endothelial surface with subsequent increased risk of thromboembolic complications. Nevertheless, there are only sparse reports on its role in the development of bleeding diathesis. Here we report two renal transplant recipients who manifested severe coagulation disorders associated with acute CMV infection. Antiviral therapy was followed by consistent correction of coagulation abnormalities. PMID- 15522111 TI - Isolated hepatic tuberculosis in a transplanted liver. AB - BACKGROUND: The incidence of tuberculosis (TB) has been increasing, especially in immunocompromised patients. It was reported that the overall incidence of TB in solid-organ transplant recipients is 0.8%. Hepatic graft TB was reported once before in a child, who underwent living-related hepatic graft transplantation. CASE REPORT: A 43-year-old man underwent orthotropic liver transplantation (OLT) in October 1999. His pre-transplant work-up was negative for TB (history and PPD skin test). He developed an episode of acute rejection which responded to steroids. He was discharged home on December 1999 with normal liver function tests (LFT). His LFT remained normal during January 2000, but his serum transaminases were found elevated on February 2000 (aspartate aminotransferase [AST] 206, alanine aminotransferase [ALT] 266). A liver biopsy then showed no evidence of TB or cytomegalovirus disease. The patient continued to have stable elevation of his serum AST and ALT until late March 2000 when a repeat liver biopsy showed caseating granuloma. The patient was started on anti-TB medications, with which he was compliant. By mid-May 2000 he was doing well, with significant reduction in his transaminase levels (AST 72, ALT 79). A retrospective inquiry about the donor revealed that he was a healthy young man from India, who died in a road traffic accident. CONCLUSION: To our knowledge our patient appears to be the first case reported of isolated hepatic TB in the OLT patient population. It is likely that the allograft was infected prior to transplantation and the disease was reactivated nearly 3 months after the procedure. PMID- 15522112 TI - A case of arthritis caused by cytomegalovirus after kidney transplantation. AB - Cytomegalovirus (CMV) is the leading cause of infectious complications after organ transplantation. We report the case of a 55-year-old renal transplant recipient who presented with CMV infection 2 months after transplantation. During oral ganciclovir treatment (5 weeks after interruption of intravenous ganciclovir), he experienced a sharp pain in the right shoulder. Examination was normal, but CMV antigenemia remained positive with 30 cells/300,000. He underwent a shoulder puncture, which confirmed the presence of CMV in the articular fluid after evaluation by polymerase chain reaction gene amplification. The patient recovered from his arthritis of the shoulder, and antigenemia became negative after 3 weeks of parenteral ganciclovir. We describe a CMV arthritis that occurred despite a curative treatment for CMV disease, and comment on the pathogenesis of this infection, the pharmalogical failure, and dosing or treatment duration. PMID- 15522113 TI - The pp67 mRNA assay in treatment and monitoring of cytomegalovirus disease in renal transplant patients in India. AB - The present report describes use of nucleic acid sequence-based amplification (NASBA) technology to detect pp67 mRNA of cytomegalovirus (CMV) in transplant patients in India. In our experience, pp67 mRNA assay was an accurate, rapid, and effective diagnostic tool to detect active CMV disease in 40.7% (50/123) of symptomatic transplant cases. This assay also allowed us to monitor CMV therapy. As part of the immunosuppressive regimen mycophenolate mofetil was found to increase the risk of developing CMV disease. All positive cases with this assay were subjected to antiviral therapy, with complete remission of the disease. At our center CMV NASBA assay has become the gold standard for the diagnosis of CMV disease in transplant patients. PMID- 15522114 TI - T-helper cell responses in liver transplant recipients: correlation with cytomegalovirus and other major infections. AB - Mitogen concanavalin A (ConA) response and cytomegalovirus (CMV)-specific memory response were assessed in 24 liver transplant recipients and compared with healthy subjects. Transplant recipients as compared to healthy subjects had a lower CMV memory response at 2 weeks (P=0.023), and at 1 month (P=0.06), but a comparable response at 3 months. CMV recipient+/donor+(R+/D+) patients had the greatest increase in CMV-specific memory response at 2-3 months as compared to all other groups. Within this R+/D+ group, CMV-specific memory response was significantly more robust in patients who never had CMV infection as compared to those who developed CMV infection (P=0.035). ConA response at 2 weeks was significantly lower in patients with major infections as compared to those without them (SI 5.4 vs. 38.1, P=0.039). Thus, reconstitution of CMV-specific T helper cell response was distinct for subsets of liver transplant recipients based on the recipient and donor CMV serostatus. Impairment in proliferative response to ConA identified a subgroup of patients with major infections after liver transplantation. PMID- 15522116 TI - Protective role of Scoparia dulcis plant extract on brain antioxidant status and lipidperoxidation in STZ diabetic male Wistar rats. AB - BACKGROUND: The aim of the study was to investigate the effect of aqueous extract of Scoparia dulcis on the occurrence of oxidative stress in the brain of rats during diabetes by measuring the extent of oxidative damage as well as the status of the antioxidant defense system. METHODS: Aqueous extract of Scoparia dulcis plant was administered orally (200 mg/kg body weight) and the effect of extract on blood glucose, plasma insulin and the levels of thiobarbituric acid reactive substances (TBARS), hydroperoxides, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced glutathione (GSH) were estimated in streptozotocin (STZ) induced diabetic rats. Glibenclamide was used as standard reference drug. RESULTS: A significant increase in the activities of plasma insulin, superoxide dismutase, catalase, glutathione peroxidase, glutathione-S-transferase and reduced glutathione was observed in brain on treatment with 200 mg/kg body weight of Scoparia dulcis plant extract (SPEt) and glibenclamide for 6 weeks. Both the treated groups showed significant decrease in TBARS and hydroperoxides formation in brain, suggesting its role in protection against lipidperoxidation induced membrane damage. CONCLUSIONS: Since the study of induction of the antioxidant enzymes is considered to be a reliable marker for evaluating the antiperoxidative efficacy of the medicinal plant, these findings suggest a possible antiperoxidative role for Scoparia dulcis plant extract. Hence, in addition to antidiabetic effect, Scoparia dulcis possess antioxidant potential that may be used for therapeutic purposes. PMID- 15522115 TI - Models of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a major global health problem and is predicted to become the third most common cause of death by 2020. Apart from the important preventive steps of smoking cessation, there are no other specific treatments for COPD that are as effective in reversing the condition, and therefore there is a need to understand the pathophysiological mechanisms that could lead to new therapeutic strategies. The development of experimental models will help to dissect these mechanisms at the cellular and molecular level. COPD is a disease characterized by progressive airflow obstruction of the peripheral airways, associated with lung inflammation, emphysema and mucus hypersecretion. Different approaches to mimic COPD have been developed but are limited in comparison to models of allergic asthma. COPD models usually do not mimic the major features of human COPD and are commonly based on the induction of COPD-like lesions in the lungs and airways using noxious inhalants such as tobacco smoke, nitrogen dioxide, or sulfur dioxide. Depending on the duration and intensity of exposure, these noxious stimuli induce signs of chronic inflammation and airway remodelling. Emphysema can be achieved by combining such exposure with instillation of tissue-degrading enzymes. Other approaches are based on genetically-targeted mice which develop COPD-like lesions with emphysema, and such mice provide deep insights into pathophysiological mechanisms. Future approaches should aim to mimic irreversible airflow obstruction, associated with cough and sputum production, with the possibility of inducing exacerbations. PMID- 15522117 TI - Discovery of mammalian genes that participate in virus infection. AB - BACKGROUND: Viruses are obligate intracellular parasites that rely upon the host cell for different steps in their life cycles. The characterization of cellular genes required for virus infection and/or cell killing will be essential for understanding viral life cycles, and may provide cellular targets for new antiviral therapies. RESULTS: Candidate genes required for lytic reovirus infection were identified by tagged sequence mutagenesis, a process that permits rapid identification of genes disrupted by gene entrapment. One hundred fifty-one reovirus resistant clones were selected from cell libraries containing 2 x 105 independently disrupted genes, of which 111 contained mutations in previously characterized genes and functionally anonymous transcription units. Collectively, the genes associated with reovirus resistance differed from genes targeted by random gene entrapment in that known mutational hot spots were under represented, and a number of mutations appeared to cluster around specific cellular processes, including: IGF-II expression/signalling, vesicular transport/cytoskeletal trafficking and apoptosis. Notably, several of the genes have been directly implicated in the replication of reovirus and other viruses at different steps in the viral lifecycle. CONCLUSIONS: Tagged sequence mutagenesis provides a rapid, genome-wide strategy to identify candidate cellular genes required for virus infection. The candidate genes provide a starting point for mechanistic studies of cellular processes that participate in the virus lifecycle and may provide targets for novel anti-viral therapies. PMID- 15522119 TI - Influence of oxygen tension on myocardial performance. Evaluation by tissue Doppler imaging. AB - BACKGROUND: Low O2 tension dilates coronary arteries and high O2 tension is a coronary vasoconstrictor but reports on O2-dependent effects on ventricular performance diverge. Yet oxygen supplementation remains first line treatment in cardiovascular disease. We hypothesized that hypoxia improves and hyperoxia worsens myocardial performance. METHODS: Seven male volunteers (mean age 38 +/- 3 years) were examined with echocardiography at respiratory equilibrium during: 1) normoxia (approximately 21% O2, 79% N2), 2) while inhaling a hypoxic gas mixture (approximately 11% O2, 89% N2), and 3) while inhaling 100% O2. Tissue Doppler recordings were acquired in the apical 4-chamber, 2-chamber, and long-axis views. Strain rate and tissue tracking displacement analyses were carried out in each segment of the 16-segment left ventricular model and in the basal, middle and apical portions of the right ventricle. RESULTS: Heart rate increased with hypoxia (68 +/- 4 bpm at normoxia vs. 79 +/- 5 bpm, P < 0.001) and decreased with hyperoxia (59 +/- 5 bpm, P < 0.001 vs. normoxia). Hypoxia increased strain rate in four left ventricular segments and global systolic contraction amplitude was increased (normoxia: 9.76 +/- 0.41 vs hypoxia: 10.87 +/- 0.42, P < 0.001). Tissue tracking displacement was reduced in the right ventricular segments and tricuspid regurgitation increased with hypoxia (7.5 +/- 1.9 mmHg vs. 33.5 +/- 1.8 mmHg, P < 0.001). The TEI index and E/E' did not change with hypoxia. Hyperoxia reduced strain rate in 10 left ventricular segments, global systolic contraction amplitude was decreased (8.83 +/- 0.38, P < 0.001 vs. normoxia) while right ventricular function was unchanged. The spectral and tissue Doppler TEI indexes were significantly increased but E/E' did not change with hyperoxia. CONCLUSION: Hypoxia improves and hyperoxia worsens systolic myocardial performance in healthy male volunteers. Tissue Doppler measures of diastolic function are unaffected by hypoxia/hyperoxia which support that the changes in myocardial performance are secondary to changes in vascular tone. It remains to be settled whether oxygen therapy to patients with heart disease is a consistent rational treatment. PMID- 15522118 TI - Thymus-derived glucocorticoids are insufficient for normal thymus homeostasis in the adult mouse. AB - BACKGROUND: It is unclear if thymus-derived glucocorticoids reach sufficient local concentrations to support normal thymus homeostasis, or if adrenal-derived glucocorticoids from the circulation are required. Modern approaches to this issue (transgenic mice that under or over express glucocorticoid receptor in the thymus) have yielded irreconcilably contradictory results, suggesting fundamental problems with one or more the transgenic mouse strains used. In the present study, a more direct approach was used, in which mice were adrenalectomized with or without restoration of circulating corticosterone using timed release pellets. Reversal of the increased number of thymocytes caused by adrenalectomy following restoration of physiological corticosterone concentrations would indicate that corticosterone is the major adrenal product involved in thymic homeostasis. RESULTS: A clear relationship was observed between systemic corticosterone concentration, thymus cell number, and percentage of apoptotic thymocytes. Physiological concentrations of corticosterone in adrenalectomized mice restored thymus cell number to normal values and revealed differential sensitivity of thymocyte subpopulations to physiological and stress-inducible corticosterone concentrations. CONCLUSION: This indicates that thymus-derived glucocorticoids are not sufficient to maintain normal levels of death by neglect in the thymus, but that apoptosis and possibly other mechanisms induced by physiological, non stress-induced levels of adrenal-derived corticosterone are responsible for keeping the total number of thymocytes within the normal range. PMID- 15522120 TI - Itch and skin rash from chocolate during fluoxetine and sertraline treatment: case report. AB - BACKGROUND: The skin contains a system for producing serotonin as well as serotonin receptors. Serotonin can also cause pruritus when injected into the skin. SSRI-drugs increase serotonin concentrations and are known to have pruritus and other dermal side effects. CASE PRESENTATION: A 46-year-old man consulted his doctor due to symptoms of depression. He did not suffer from any allergy but drinking red wine caused vasomotor rhinitis. Antidepressive treatment with fluoxetine 20 mg daily was initiated which was successful. After three weeks of treatment an itching rash appeared. An adverse drug reaction (ADR) induced by fluoxetine was suspected and fluoxetine treatment was discontinued. The symptoms disappeared with clemastine and betametasone treatment. Since the depressive symptoms returned sertraline medication was initiated. After approximately two weeks of sertraline treatment he noted an intense itching sensation in his scalp after eating a piece of chocolate cake. The itch spread to the arms, abdomen and legs and the patient treated himself with clemastine and the itch disappeared. He now realised that he had eaten a chocolate cake before this episode and remembered that before the first episode he had had a chocolate mousse dessert. He had never had any reaction from eating chocolate before and therefore reported this observation to his doctor. CONCLUSIONS: This case report suggests that there may be individuals that are very sensitive to increases in serotonin concentrations. Dermal side reactions to SSRI-drugs in these patients may be due to high activity in the serotonergic system at the dermal and epidermo-dermal junctional area rather than a hypersensitivity to the drug molecule itself. PMID- 15522121 TI - An information integration theory of consciousness. AB - BACKGROUND: Consciousness poses two main problems. The first is understanding the conditions that determine to what extent a system has conscious experience. For instance, why is our consciousness generated by certain parts of our brain, such as the thalamocortical system, and not by other parts, such as the cerebellum? And why are we conscious during wakefulness and much less so during dreamless sleep? The second problem is understanding the conditions that determine what kind of consciousness a system has. For example, why do specific parts of the brain contribute specific qualities to our conscious experience, such as vision and audition? PRESENTATION OF THE HYPOTHESIS: This paper presents a theory about what consciousness is and how it can be measured. According to the theory, consciousness corresponds to the capacity of a system to integrate information. This claim is motivated by two key phenomenological properties of consciousness: differentiation - the availability of a very large number of conscious experiences; and integration - the unity of each such experience. The theory states that the quantity of consciousness available to a system can be measured as the Phi value of a complex of elements. Phi is the amount of causally effective information that can be integrated across the informational weakest link of a subset of elements. A complex is a subset of elements with Phi>0 that is not part of a subset of higher Phi. The theory also claims that the quality of consciousness is determined by the informational relationships among the elements of a complex, which are specified by the values of effective information among them. Finally, each particular conscious experience is specified by the value, at any given time, of the variables mediating informational interactions among the elements of a complex. TESTING THE HYPOTHESIS: The information integration theory accounts, in a principled manner, for several neurobiological observations concerning consciousness. As shown here, these include the association of consciousness with certain neural systems rather than with others; the fact that neural processes underlying consciousness can influence or be influenced by neural processes that remain unconscious; the reduction of consciousness during dreamless sleep and generalized seizures; and the time requirements on neural interactions that support consciousness. IMPLICATIONS OF THE HYPOTHESIS: The theory entails that consciousness is a fundamental quantity, that it is graded, that it is present in infants and animals, and that it should be possible to build conscious artifacts. PMID- 15522122 TI - Mercury exposure, malaria, and serum antinuclear/antinucleolar antibodies in Amazon populations in Brazil: a cross-sectional study. AB - BACKGROUND: Mercury is an immunotoxic metal that induces autoimmune disease in rodents. Highly susceptible mouse strains such as SJL/N, A.SW, B10.S (H-2s) develop multiple autoimmune manifestations after exposure to inorganic mercury, including lymphoproliferation, elevated levels of autoantibodies, overproduction of IgG and IgE, and circulating immune complexes in kidney and vasculature. A few studies have examined relationships between mercury exposures and adverse immunological reactions in humans, but there is little evidence of mercury associated autoimmunity in humans. METHODS: To test the immunotoxic effects of mercury in humans, we studied communities in Amazonian Brazil with well characterized exposures to mercury. Information was collected on diet, mercury exposures, demographic data, and medical history. Antinuclear and antinucleolar autoantibodies (ANA and ANoA) were measured by indirect immunofluorescence. Anti fibrillarin autoantibodies (AFA) were measured by immunoblotting. RESULTS: In a gold mining site, there was a high prevalence of ANA and ANoA: 40.8% with detectable ANoA at > or =1:10 serum dilution, and 54.1% with detectable ANA (of which 15% had also detectable ANoA). In a riverine town, where the population is exposed to methylmercury by fish consumption, both prevalence and levels of autoantibodies were lower: 18% with detectable ANoA and 10.7% with detectable ANA. In a reference site with lower mercury exposures, both prevalence and levels of autoantibodies were much lower: only 2.0% detectable ANoA, and only 7.1% with detectable ANA. In the gold mining population, we also examined serum for AFA in those subjects with detectable ANoA (> or =1:10). There was no evidence for mercury induction of this autoantibody. CONCLUSIONS: This is the first study to report immunologic changes, indicative of autoimmune dysfunction in persons exposed to mercury, which may also reflect interactions with infectious disease and other factors. PMID- 15522124 TI - Metabolomics: an emerging post-genomic tool for nutrition. AB - The post-genomic era has been driven by the development of technologies that allow the function of cells and whole organisms to be explored at the molecular level. Metabolomics is concerned with the measurement of global sets of low molecular-weight metabolites. Metabolite profiles of body fluids or tissues can be regarded as important indicators of physiological or pathological states. Such profiles may provide a more comprehensive view of cellular control mechanisms in man and animals, and raise the possibility of identifying surrogate markers of disease. Metabolomic approaches use analytical techniques such as NMR spectroscopy and MS to measure populations of low-molecular-weight metabolites in biological samples. Advanced statistical and bioinformatic tools are then employed to maximise the recovery of information and interpret the large datasets that are generated. Metabolomics has already been used to study toxicological mechanisms and disease processes and offers enormous potential as a means of investigating the complex relationship between nutrition and metabolism. Examples include the metabolism of dietary substrates, drug-induced disturbances of lipid metabolites in type 2 diabetes mellitus and the therapeutic effects of vitamin supplementation in the treatment of chronic metabolic disorders. PMID- 15522123 TI - Acetylation of insulin receptor substrate-1 is permissive for tyrosine phosphorylation. AB - BACKGROUND: Insulin receptor substrate (IRS) proteins are key moderators of insulin action. Their specific regulation determines downstream protein-protein interactions and confers specificity on growth factor signalling. Regulatory mechanisms that have been identified include phosphorylation of IRS proteins on tyrosine and serine residues and ubiquitination of lysine residues. This study investigated other potential molecular mechanisms of IRS-1 regulation. RESULTS: Using the sos recruitment yeast two-hybrid system we found that IRS-1 and histone deacetylase 2 (HDAC2) interact in the cytoplasmic compartment of yeast cells. The interaction mapped to the C-terminus of IRS-1 and was confirmed through co immunoprecipitation in vitro of recombinant IRS-1 and HDAC2. HDAC2 bound to IRS-1 in mammalian cells treated with phorbol ester or after prolonged treatment with insulin/IGF-1 and also in the livers of ob/ob mice but not PTP1B knockout mice. Thus, the association occurs under conditions of compromised insulin signalling. We found that IRS-1 is an acetylated protein, of which the acetylation is increased by treatment of cells with Trichostatin A (TSA), an inhibitor of HDAC activity. TSA-induced increases in acetylation of IRS-1 were concomitant with increases in tyrosine phosphorylation in response to insulin. These effects were confirmed using RNA interference against HDAC2, indicating that HDAC2 specifically prevents phosphorylation of IRS-1 by the insulin receptor. CONCLUSIONS: Our results show that IRS-1 is an acetylated protein, a post translational modification that has not been previously described. Acetylation of IRS-1 is permissive for tyrosine phosphorylation and facilitates insulin stimulated signal transduction. Specific inhibition of HDAC2 may increase insulin sensitivity in otherwise insulin resistant conditions. PMID- 15522125 TI - The use of high-selenium yeast to raise selenium status: how does it measure up? AB - Selenium-enriched yeast (Se-yeast) is a common form of Se used to supplement the dietary intake of this important trace mineral. However, its availability within the European Union is under threat, owing to concerns expressed by the European Community (EC) Scientific Committee on Food that Se-yeast supplements are poorly characterised and could potentially cause the build up of Se in tissues to toxic levels. The present review examines the validity of these concerns. Diagrams of the biosynthesis and metabolism of Se compounds show which species can be expected to occur in Se-yeast preparations. Se-yeast manufacture is described together with quality-control measures applied by reputable manufacturers. The way in which speciation of Se-yeast is achieved is explained and results on amounts of Se species in various commercial products are tabulated. In all cases described, selenomethionine is the largest single species, accounting for 54-74 % of total Se. Se-yeast is capable of increasing the activity of the selenoenzymes and its bioavailability has been found to be higher than that of inorganic Se sources in all but one study. Intervention studies with Se-yeast have shown the benefit of this form in cancer prevention, on the immune response and on HIV infection. Of about one dozen supplementation studies, none has shown evidence of toxicity even up to an intake level of 800 microg Se/d over a period of years. It is concluded that Se-yeast from reputable manufacturers is adequately characterised, of reproducible quality, and that there is no evidence of toxicity even at levels far above the EC tolerable upper intake level of 300 microg/d. PMID- 15522126 TI - The absorption of vitamin E is influenced by the amount of fat in a meal and the food matrix. AB - Vitamin E absorption requires the presence of fat; however, limited information exists on the influence of fat quantity on optimal absorption. In the present study we compared the absorption of stable-isotope-labelled vitamin E following meals of varying fat content and source. In a randomised four-way cross-over study, eight healthy individuals consumed a capsule containing 150 mg (2)H labelled RRR-alpha-tocopheryl acetate with a test meal of toast with butter (17.5 g fat), cereal with full-fat milk (17.5 g fat), cereal with semi-skimmed milk (2.7 g fat) and water (0 g fat). Blood was taken at 0, 0.5, 1, 1.5, 2, 3, 6 and 9 h following ingestion, chylomicrons were isolated, and (2)H-labelled alpha tocopherol was analysed in the chylomicron and plasma samples. There was a significant time (P<0.001) and treatment effect (P<0.001) in (2)H-labelled alpha tocopherol concentration in both chylomicrons and plasma between the test meals. (2)H-labelled alpha-tocopherol concentration was significantly greater with the higher-fat toast and butter meal compared with the low-fat cereal meal or water (P<0.001), and a trend towards greater concentration compared with the high-fat cereal meal (P=0.065). There was significantly greater (2)H-labelled alpha tocopherol concentration with the high-fat cereal meal compared with the low-fat cereal meal (P<0.05). The (2)H-labelled alpha-tocopherol concentration following either the low-fat cereal meal or water was low. These results demonstrate that both the amount of fat and the food matrix influence vitamin E absorption. These factors should be considered by consumers and for future vitamin E intervention studies. PMID- 15522127 TI - Protein quantity, not protein quality, accelerates whole-body leucine kinetics and the acute-phase response during acute infection in marasmic Malawian children. AB - The present study compared leucine kinetics and acute-phase-protein concentrations in three groups of marasmic, acutely infected Malawian children fed one of three isoenergetic diets. These were: an enhanced-protein-quality diet (egg-white+tryptophan, providing 1.2 g protein/kg per d; n 14); an increased protein-content diet (egg-white+tryptophan, providing 1.8 g protein/kg per d; n 14); a standard-protein diet (1.2 g milk protein/kg per d; n 25). The hypotheses tested were that children receiving a diet with more protein would have greater rates of non-oxidative leucine disposal and that children receiving an isonitrogenous diet with a higher protein quality would have lower rates of leucine oxidation. The children were studied after 24 h of therapy using standard [(13)C]leucine stable-isotope tracer techniques. The children receiving the higher-protein-content diet had greater leucine kinetic rates than those receiving the standard-protein-content diet; non-oxidative leucine disposal was 170 (sd 52) v. 122 (sd 30) mumol leucine/kg per h (P<0.01). Leucine oxidation was less in the children receiving the enhanced-protein-quality diet than in those receiving the standard-protein-quality diet; 34 (sd 12) v. 45 (sd 13) mumol leucine/kg per h (P<0.05). The children receiving the high-protein-content diet increased their serum concentration for five of six acute-phase proteins 24 h after starting therapy, while those receiving the standard-protein-content diet did not. These data suggest that there was greater whole-body protein synthesis, and a more vigorous acute-phase response associated with the higher-protein content diet. The clinical benefits associated with a higher protein intake in marasmic, acutely infected children need further study. PMID- 15522128 TI - Flupenthixol and cefotiam: effects on vitamin A metabolism in rats. AB - We examined the alterations in vitamin A metabolism as a result of flupenthixol or cefotiam administration. The impact of these drugs on indices of vitamin A status was evaluated in Brown Norway and Long-Evans rats. Intramuscular drug administration for 28 d resulted in a decline in systemic retinol. Changes in circulating retinol with time for chronic dosing showed drug treatment (P<0.001) and time (P<0.03) to be significant factors, but rat strain (P=0.33) was not a significant factor. Flupenthixol was the most active retinol-lowering compound (P<0.005). At the end of the 28 d period, hepatic retinyl ester hydrolase activity was greater in drug-treated rats than in controls (P<0.05). With regard to effects on liver reserves: (1) flupenthixol treatment resulted in vitamin A depletion (P<0.05); (2) cefotiam treatment stimulated vitamin A accumulation; (3) distinctive patterns of retinol and its esters were seen in response to treatment. It is reasonable to assume that the drugs interfere with vitamin A in at least two ways: (1) lowering of plasma retinol, an early event in the interaction, may be caused by inhibition of hepatic holo-retinol-binding protein secretion or stimulation of clearance, or both; (2) when plasma retinol levels are persistently low, and as the hepatic deposits of the xenobiotics build up, there are changes in the vitamin A pool size and composition of the liver. Candidate enzymes are retinyl ester hydrolase and cytochrome P450. The relationship between these two events will be studied in further detail. PMID- 15522129 TI - A novel high-amylose barley cultivar (Hordeum vulgare var. Himalaya 292) lowers plasma cholesterol and alters indices of large-bowel fermentation in pigs. AB - Hordeum vulgare var. Himalaya 292 is a new barley cultivar with altered starch synthesis and less total starch but more amylose, resistant starch (RS) and total and soluble NSP including beta-glucan. To determine its nutritional potential, young pigs were fed diets containing stabilised wholegrain flours from either Himalaya 292, Namoi (a commercial barley), wheat bran or oat bran at equivalent dietary NSP concentrations for 21 d. Serum total cholesterol was significantly lowered by the Himalaya 292 diet relative to wheat bran, indicating that Himalaya 292 retained its hypocholesterolaemic potential. In all groups SCFA concentrations were highest in the proximal colon and decreased towards the rectum. Digesta pH was lowest in the proximal colon and highest in the distal colon. Large-bowel and faecal pH were significantly lower in the pigs fed the barley and oat diets, indicating greater bacterial fermentation. Caecal and proximal colonic pH was lowest and SCFA pools highest in the pigs fed Himalaya 292. Total and individual SCFA were lowest in the mid- and distal colon of the pigs fed Himalaya 292 or oat bran. These data suggest the presence of more RS in Himalaya 292 and suggest that its fermentation was rapid relative to transit. Differences in faecal and large-bowel anaerobic, aerobic, coliform and lactic acid bacteria were relatively small, indicating a lack of a specific prebiotic action. These data support the potential of this novel barley cultivar to improve health through plasma cholesterol reduction and increased large-bowel SCFA production. PMID- 15522130 TI - Nutritional status induces divergent variations of GLUT4 protein content, but not lipoprotein lipase activity, between adipose tissues and muscles in adult cattle. AB - Metabolic adaptations to variations in food supply are incompletely understood in ruminant animal adipose tissue (AT) and muscle. To explore this, we studied lipid metabolism and glucose transport potential in one internal and one external AT, as well as in one oxidative and one glycolytic muscle from control, 7 d underfed and 21 d refed adult cows. Refeeding increased (+79 to +307 %) the activities of enzymes involved in de novo lipogenesis (fatty acid synthase, malic enzyme, glucose-6-phosphate dehydrogenase) in perirenal and subcutaneous AT; underfeeding did not modify these variables. Underfeeding decreased the activities of lipoprotein lipase (LPL) in perirenal AT (-70 %) and cardiac muscle (-67 %), but did not modify the activities in subcutaneous AT and longissimus thoracis. Refeeding increased LPL activities in all tissues (+40 to +553 %) to levels comparable with (cardiac muscle) or greater than (AT, longissimus thoracis) those observed in control cows. Such variations in perirenal and cardiac muscle LPL activities did not result from variations in LPL mRNA levels, but suggest a post transcriptional regulation of LPL in these nutritional conditions. Underfeeding did not modify GLUT4 contents in perirenal AT and muscles, while refeeding increased it only in perirenal AT (+250 %). Our present results contrast with previous results in rats, where LPL is regulated in opposite directions in AT and muscles, and GLUT4 is generally increased by fasting and decreased by refeeding in skeletal muscles. The present results highlight the bovine specificity of the response, which probably arises in part from peculiarities of ruminant animals for nutrient digestion and absorption. PMID- 15522131 TI - Dose dependency of the effect of ornithine alpha-ketoglutarate on tissue glutamine concentrations and hypercatabolic response in endotoxaemic rats. AB - The optimal dosage of ornithine alpha-ketoglutarate (OKG) for repleting tissue glutamine (Gln) concentrations and maintaining N homeostasis after injury is unknown. We set out to perform 'dose-ranging' of OKG supplementation after an endotoxaemic challenge. Sixty-one male Wistar rats were injected with 3 mg lipopolysaccharide (LPS) from Escherichia coli/kg (n 50) or saline vehicle (9 g NaCl/l; controls n 11). After a 24 h fast, survivors were fed by gavage for 48 h with a polymeric standard diet (879 kJ/kg per d and 1.18 g N/kg per d) supplemented with non-essential amino acids (control, n 11; LPS-OKG-0.0, n 9), or with 0.5 g OKG/kg per d (LPS-OKG-0.5, n 12), 1.5 OKG/kg per d (LPS-OKG-1.5, n 11) or 4.5 g OKG/kg per d (LPS-OKG-4.5, n 10). The diets for all groups were made isonitrogenous with the LPS-OKG-4.5 diet by adding an appropriate amount of non essential amino acids. Rats were killed on day 3 for blood and tissue sampling (muscle, jejunum mucosa, liver). Urine was collected daily for 3-methylhistidine and total N assays. The OKG dose was correlated with Gln concentrations in every tissue and with cumulative N balance (Spearman test, P<0.01). 3-Methylhistidine excretion was increased in endotoxaemic groups compared with controls (ANOVA, P<0.05) except in the LPS-OKG-4.5 group. Only the LPS-OKG-4.5 group achieved a positive post-injury N balance (t test, P<0.05). In conclusion, OKG exerted a dose-dependent effect on tissue Gln concentration and N balance, but only the highest dosage counteracted myofibrillar hypercatabolism and caused a positive N balance. PMID- 15522132 TI - Potential vasorelaxant effects of oleanolic acid and erythrodiol, two triterpenoids contained in 'orujo' olive oil, on rat aorta. AB - 'Orujo' olive oil is obtained by chemical processes from the waste resulting from the mechanical extraction of virgin olive oil. The aim of the present study was to evaluate a new pharmacological property of two natural triterpenoids contained in olive oil, as vasodilatory agents, and to determine their mechanism of action. The two compounds studied were oleanolic acid and erythrodiol. The vasorelaxant effect induced by these pentacyclic triterpenoids was studied in isolated thoracic rat aorta. Oleanolic acid and erythrodiol, accumulatively added, showed vasorelaxant activities in aortic rings with endothelium pre-contracted by 10(-6) m-phenylephrine (maximum percentage of relaxation 86.38 (sem 2.89) and 73.53 (sem 6.01), respectively). They had almost no relaxant effect on depolarised or endothelium-denuded aortic segments. The relaxation was significantly attenuated by pre-treatment with the NO synthase inhibitor N(omega)-nitro-L-arginine methylester (L-NAME; 3x10(-4) m). To characterise the involvement of endothelial factors, in addition to NO, arteries with endothelium were exposed to 10(-5) m indomethacin (INDO), a cyclo-oxygenase inhibitor, or INDO plus L-NAME. INDO did not have any significant effect on the relaxant response of both compounds. The combination of L-NAME plus INDO only abolished the oleanolic acid-induced relaxation. The present results suggest that the mechanism of relaxation seems to be mainly mediated by the endothelial production of NO; however, other mechanisms cannot be excluded. It can be concluded that oleanolic acid and erythrodiol may have interesting therapeutic potential as new vasodilator drugs, thus protecting the cardiovascular system. Therefore, the intake of 'orujo' olive oil, as a source of these compounds, might be beneficial in this regard. PMID- 15522133 TI - Dietary n-6:n-3 fatty acid ratio in the perinatal period affects bone parameters in adult female rats. AB - PUFA and their metabolites are important regulators of bone formation and resorption. The effect of PUFA on bone growth may be especially striking during the perinatal period. The aim of the present study was to investigate the effect of diets with different n-6:n-3 fatty acid (FA) ratios during the perinatal period on bone parameters in the adult offspring. During late gestation and throughout lactation, rat dams were fed an isoenergetic diet containing 70 g linseed oil (n-3 diet), soyabean oil (n-6+n-3 diet) or sunflower-seed oil (n-6 diet) per kg with n-6:n-3 FA ratios of 0.4, 9 and 216, respectively. The offspring were weaned onto an ordinary chow and followed until 30 weeks of age. Bone parameters were analysed using peripheral quantitative computerised tomography and dual-energy X-ray absorptiometry. Femur length and cortical cross sectional bone area and bone mineral content were significantly higher in the n 6+n-3 group than in the other groups. Cortical bone thickness in the n-6+n-3 group was increased compared with the n-3 group, but most cortical bone parameters did not differ between the n-3 and n-6 groups. The results suggest that regulatory mechanisms were influenced by the n-6:n-3 FA ratio early in life and not compensated for by the introduction of an ordinary diet after weaning. PMID- 15522134 TI - Background diet influences the anti-inflammatory effect of alpha-linolenic acid in dyslipidaemic subjects. AB - Long-chain n-3 PUFA from fish oils are known to have anti-inflammatory effects. We evaluated the effect of alpha-linolenic acid (ALA), precursor of n-3 fatty acids, on serum inflammatory markers and soluble cellular adhesion molecules (sCAM) of dyslipidaemic males, relative to their background diet. Participants were assigned to two groups, based upon food intake patterns: (a) twenty-one dyslipidaemic subjects who habitually ate a Mediterranean-Cretan-type diet; (b) nineteen dyslipidaemic subjects who normally ate a Westernised Greek diet. All were supplemented with 8.1 g ALA/d for 12 weeks. We determined serum amyloid A (SAA), C-reactive protein (CRP), macrophage colony-stimulating factor (MCSF), IL 6, soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 and soluble E-selectin concentrations at the beginning and the end of the ALA supplementation period. Serum baseline concentrations of inflammatory markers and sCAM were similar across the diet groups. Type of diet had a significant impact on the response of inflammatory markers to ALA supplementation. The Westernised Greek diet group showed a reduction in SAA (P<0.001), CRP (P=0.002), MCSF (P=0.005) and IL-6 (P=0.04) concentrations. The Mediterranean-Cretan-type background diet group showed a significant reduction only in MCSF concentrations (P=0.003). The sVCAM-1 concentrations were significantly reduced in both the Westernised Greek diet group (P=0.001) and the Mediterranean-Cretan-type diet group (P<0.001). The present study demonstrated that ALA supplementation lowered the serum concentrations of inflammatory markers more profoundly when the background diet was rich in saturated fatty acids and poor in MUFA. PMID- 15522135 TI - Combined effects of a dietary portfolio of plant sterols, vegetable protein, viscous fibre and almonds on LDL particle size. AB - Studies conducted in the last 20 years have led to the identification of small dense LDL as an important risk factor for CVD. Consumption of plant sterols, soyabean proteins, viscous fibre and nuts are known to modulate the risk of CVD favourably through their cholesterol (Chol)-lowering properties, both independently and more recently in combination. Nevertheless, their combined impact on the LDL particle size phenotype has never been tested. In the present study, we assessed the effect of incorporating concurrently plant sterols (1 g/4.2 MJ), soyabean protein (23 g/4.2 MJ), viscous fibre (9 g/4.2 MJ) and almonds (15 g/4.2 MJ) into a diet very low in saturated fat in twelve patients with mildly elevated plasma LDL-Chol levels. Fasting blood lipids were obtained at the start of the study and at 2-week intervals during the 4-week study. The diet induced reduction in plasma LDL-Chol of 30.0 (se 3.0) % (P<0.0001) was attributed to concurrent reductions in the serum Chol concentrations of large (>26.0 nm-30 (se 8) %, P<0.001), medium (25.5-26.0 nm-29 (se 3) %, P<0.001) and small (<25.5 nm-21 (sd 6) %, P<0.01) LDL particles, with near maximal reductions seen by week 2. These results indicate that foods and dietary components advocated for their potential to reduce the risk of CVD are effective in reducing serum concentrations of all LDL fractions including small dense LDL, thus potentially further contributing to an overall lower risk of CVD. PMID- 15522136 TI - The effect of low doses of betaine on plasma homocysteine in healthy volunteers. AB - Homocysteine is a risk factor for vascular diseases, and lowering of plasma total homocysteine (tHcy) may be beneficial for health. Homocysteine can be remethylated to methionine by betaine-homocysteine methyltransferase using betaine (2(N,N,N-trimethyl)glycine) as methyl donor. A dose of 6 g betaine/d has been used in the treatment of homocystinuria, but data on the dose-response are scarce. Thirty-four healthy men and women were supplied with doses of 1, 3 and 6 g betaine and then with 6 g betaine+1 mg folic acid for four consecutive 1-week periods. The mean plasma tHcy concentration decreased by 1.1 (NS), 10.0 and 14.0 % (P<0.001) after supplementation with 1, 3 and 6 g betaine respectively. A further decrease in plasma tHcy by 5 % (P<0.01) was achieved by combining 1 mg folic acid with the 6 g betaine dose. Plasma betaine increased from 31 (sd 13) to 255 (sd 136) mumol/l in a dose-dependent manner (R(2) 0.97). We conclude that plasma tHcy is lowered rapidly and significantly by 3 or 6 g betaine/d in healthy men and women. PMID- 15522137 TI - Estimation of plant sterol and cholesterol intake in Finland: quality of new values and their effect on intake. AB - The Finnish national food composition database Fineli was updated with recent analytical values for plant sterols (PS) (sitosterol, campesterol, stigmasterol, avenasterol, brassicasterols and stanols) and cholesterol. The quality of the new analytical data was assessed. The aims of the present study were: (1) to compare the effect of old and new database values on PS and cholesterol intakes based on average per capita food consumption data; (2) to estimate the current intake and major sources of these compounds in various population groups according to the national FINDIET 1997 survey data. The intake of total PS was 305 mg/d for men and 237 mg/d for women. The respective intakes for cholesterol were 284 mg/d and 201 mg/d. Women had a higher density of PS in their diets than men, whereas the cholesterol density in the diets did not differ between genders. Cereals, margarine, vegetables and vegetable oils were the main food sources of PS. Meat, meat products and eggs were the main sources of cholesterol. A 9 % greater PS intake estimate was obtained with the new PS database compared with the old PS database, probably due to minor methodological differences between the new and old analyses. Notable changes in analytical methods suggest a lower value (-19 %) for cholesterol intake calculated from the new database compared with the old one. We conclude that researchers can have confidence in the new values for PS and cholesterol, because systematic evaluation of the new analytical values showed them to be of high quality. PMID- 15522138 TI - Trends in energy and macronutrient intakes, body weight and physical activity in female university students (1988-2003), and effects of excluding under-reporters. AB - The present study investigated trends in reported energy intake, macronutrient intake, physical activity level (PAL) and body weight and effects of excluding under-reporters (UR). Dietary intake and time spent in sixteen activity categories were recorded by 887 female university students (median age 29 years) from 1988 to 2003. Energy expenditure (EE) and PAL were measured using a factorial method. All data collected were self-reported. Individuals with reported EI:EE<0.76 were classified as UR. The remainder were classified as non under-reporters (NUR). Trends were determined from simple linear regression of median data for each year for the entire cohort (ALL) and for NUR and UR separately, and from multiple regression analysis with the subgroups (NUR and UR) as an additional predictor (BOTH). Prevalence of under-reporting and overweight increased between 1988 and 2003. In ALL and BOTH there were trends to increased body mass, protein intake (g/d and % energy) and carbohydrate intake (% energy only) and decreased fat and alcohol intakes (g/d and % energy). In BOTH there were also increases in reported EI and carbohydrate intake (g/d). None of the trends in NUR was significantly different from those in UR, but some trends in ALL and/or BOTH were not significant when UR were excluded. Trends remaining significant in NUR were increased reported energy intake, protein (g/d) and carbohydrate (g/d) intakes, and decreased fat (% energy only) intake. There were no significant trends in PAL. We conclude that some, but not all, dietary trends were affected by exclusion of UR. PMID- 15522139 TI - Rumen simulation technique study on the interactions of dietary lauric and myristic acid supplementation in suppressing ruminal methanogenesis. AB - The interactions of lauric (C12) and myristic acid (C14) in suppressing ruminal methanogenesis and methanogens were investigated with the rumen simulation technique (Rusitec) using bovine ruminal fluid. The fatty acids were added to basal substrates (grass hay:concentrate, 1:1.5) at a level of 48 g/kg DM, provided in C12:C14 ratios of 5:0, 4:1, 3:2, 2.5:2.5, 2:3, 1:4 and 0:5. Additionally, an unsupplemented control consisting of the basal substrates only was employed. Incubation periods lasted for 15 (n 4) and 25 (n 2) d. CH4 formation was depressed by any fatty acid mixture containing at least 40 % C12, and effects persisted over the complete incubation periods. The greatest depression (70 % relative to control) occurred with a C12:C14 ratio of 4:1, whereas the second most effective treatment in suppressing CH4 production (60 % relative to control) was found with a ratio of 3:2. Total methanogenic counts were decreased by those mixtures of C12 and C14 also successful in suppressing methanogenesis, the 4:1 treatment being most efficient (60 % decline). With this treatment in particular, the composition of the methanogenic population was altered in such a way that the proportion of Methanococcales increased and Methanobacteriales decreased. Initially, CH4 suppression was associated with a decreased fibre degradation, which, however, was reversed after 10 d of incubation. The present study demonstrated a clear synergistic effect of mixtures of C12 and C14 in suppressing methanogenesis, mediated probably by direct inhibitory effects of the fatty acids on the methanogens. PMID- 15522140 TI - Effect of garlic (Allium sativum) powder tablets on serum lipids, blood pressure and arterial stiffness in normo-lipidaemic volunteers: a randomised, double blind, placebo-controlled trial. AB - Recent studies have cast doubt on the proposed lipid-lowering and blood pressure lowering effects of garlic. We tested the effect of dried garlic (Allium sativum) powder on blood lipids, blood pressure and arterial stiffness in a 12-week randomised, double-blind, placebo-controlled trial. Seventy-five healthy, normo lipidaemic volunteers (men and women aged 40-60 years) were assigned to dried garlic powder tablets (10.8 mg alliin (3-(2-propenylsulfinyl)-L-alanine)/d, corresponding to about three garlic cloves) or placebo. Sixty-two subjects were eligible for the per-protocol analysis. The primary outcome measure was serum total cholesterol concentration. Secondary outcome measures were LDL-cholesterol, HDL-cholesterol and triacylglycerol concentrations, blood pressure and arterial stiffness (assessed by pulse wave velocity). No significant differences between the garlic and placebo groups were detected for any of the outcome measures. However, garlic powder was associated with a near-significant decrease (12 %) in triacylglycerol concentration (P=0.07). In conclusion, garlic powder tablets have no clinically relevant lipid-lowering and blood pressure-lowering effects in middle-aged, normo-lipidaemic individuals. The putative anti-atherosclerotic effect of garlic may be linked to risk markers other than blood lipids. PMID- 15522141 TI - Modelling of nutrient partitioning in growing pigs to predict their anatomical body composition. 1. Model description. AB - A dynamic mechanistic model was developed for growing and fattening pigs. The aim of the model was to predict growth rate and the chemical and anatomical body compositions from the digestible nutrient intake of gilts (20-105 kg live weight). The model represents the partitioning of digestible nutrients from intake through intermediary metabolism to body protein and body fat. State variables of the model were lysine, acetyl-CoA equivalents, glucose, volatile fatty acids and fatty acids as metabolite pools, and protein in muscle, hide backfat, bone and viscera and body fat as body constituent pools. It was assumed that fluxes of metabolites follow saturation kinetics depending on metabolite concentrations. In the model, protein deposition rate depended on the availability of lysine and of acetyl-CoA. The anatomical body composition in terms of muscle, organs, hide-backfat and bone was predicted from the chemical body composition and accretion using allometric relationships. Partitioning of protein, fat, water and ash in muscle, organs, hide-backfat and bone fractions were driven by the rates of muscle protein and body fat deposition. Model parameters were adjusted to obtain a good fit of the experimental data from literature. Differential equations were solved numerically for a given set of initial conditions and parameter values. In the present paper, the model is presented, including its parameterisation. The evaluation of the model is described in a companion paper. PMID- 15522142 TI - Modelling of nutrient partitioning in growing pigs to predict their anatomical body composition. 2. Model evaluation. AB - The objective of the present paper was to evaluate a dynamic mechanistic model for growing and fattening pigs presented in a companion paper. The model predicted the rate of protein and fat deposition (chemical composition), rate of tissue deposition (anatomical composition) and performance of pigs depending on nutrient intake. In the model evaluation, the predicted response of the pig to changes in model parameters and to changes in nutrient intakes is presented. As a result of the sensitivity analysis, changes in the maintenance energy requirements and the fractional degradation rate of muscle protein had the greatest impact on tissue deposition rates. The model was also highly sensitive to changes in the maximum velocity and steepness parameter of the lysine utilisation for muscle protein synthesis. The model was further tested by independent published results. The model successfully predicted the response of pigs to a wide range of variation in nutrient composition. Consequently, the model can be applied to develop feeding strategies to optimise pig production. It also enables prediction of the slaughter performance and the meat quality. PMID- 15522143 TI - Food and drinking patterns as predictors of 6-year BMI-adjusted changes in waist circumference. AB - Few studies have investigated the prospective associations between diet or drinking patterns and abdominal obesity; we therefore investigated whether food and beverage groups or patterns predicted 6-year changes in waist circumference (WC) and whether these associations were independent of concurrent changes in BMI as a measure of general obesity. The subjects were 2300 middle-aged men and women with repeated measurements of dietary intake, BMI and WC from 1982 to 1993. Intakes from ten food groups and from coffee, tea, wine, beer and spirits were assessed; gender-specific food factors were identified by factor analyses. Multiple linear regression analyses were done before and after adjustment for concurrent changes in BMI. A high intake of potatoes seemed to prevent gain in WC for men, while a high intake of refined bread was associated with gain in WC for women. The association persisted for refined bread, but not for potatoes, after adjustment for concurrent BMI changes. Among women, but not men, high intakes of beer and spirits were associated with gain in WC in both models. A high intake of coffee for women and moderate to high intake of tea for men were associated with gain in WC, but the associations were weakened, especially for women, after adjustment for BMI changes. None of the food factors was associated with WC changes. Based on the present study, we conclude that very few food items and no food patterns seem to predict changes in WC, whereas high intakes of beer and spirits among women, and moderate to high tea intake among men, may promote gain in WC. PMID- 15522158 TI - Dietary intake and nutritional status of children and adolescents in Europe. AB - The objective of this project was to collect and evaluate data on nutrient intake and status across Europe and to ascertain whether any trends could be identified. Surveys of dietary intake and status were collected from across Europe by literature search and personal contact with country experts. Surveys that satisfied a defined set of criteria -- published, based on individual intakes, post-1987, adequate information provided to enable its quality to be assessed, small age bands, data for sexes separated above 12 years, sample size over 25 and subjects representative of the population -- were selected for further analysis. In a small number of cases, where no other data for a country were available or where status data were given, exceptions were made. Seventy-nine surveys from 23 countries were included, and from them data on energy, protein, fats, carbohydrates, alcohol, vitamins, minerals and trace elements were collected and tabulated. Data on energy, protein, total fat and carbohydrate were given in a large number of surveys, but information was very limited for some micronutrients. No surveys gave information on fluid intake and insufficient gave data on food patterns to be of value to this project. A variety of collection methods were used, there was no consistency in the ages of children surveyed or the age cut-off points, but most surveys gave data for males and females separately at all ages. Just under half of the surveys were nationally representative and most of the remainder were regional. Only a small number of local surveys could be included. Apart from anthropometric measurements, status data were collected in only seven countries. Males had higher energy intakes than females, energy intake increased with age but levelled off in adolescent girls. Intakes of other nutrients generally related to energy intakes. Some north-south geographical trends were noted in fat and carbohydrate intakes, but these were not apparent for other nutrients. Some other trends between countries were noted, but there were also wide variations within countries. A number of validation studies have shown that misreporting is a major problem in dietary surveys of children and adolescents and so all the dietary data collected for this project should be interpreted and evaluated with caution. In addition, dietary studies rely on food composition tables for the conversion of food intake data to estimated nutrient intakes and each country uses a different set of food composition data which differ in definitions, analytical methods, units and modes of expression. This can make comparisons between countries difficult and inaccurate. Methods of measuring food intake are not standardised across Europe and intake data are generally poor, so there are uncertainties over the true nutrient intakes of children and adolescents across Europe. There are insufficient data on status to be able to be able to draw any conclusions about the nutritional quality of the diets of European children and adolescents. PMID- 15522159 TI - Issues in dietary intake assessment of children and adolescents. AB - Studies of food habits and dietary intakes face a number of unique respondent and observer considerations at different stages from early childhood to late adolescence. Despite this, intakes have often been reported as if valid, and the interpretation of links between intake and health has been based, often erroneously, on the assumption of validity. However, validation studies of energy intake data have led to the widespread recognition that much of the dietary data on children and adolescents is prone to reporting error, mostly through under reporting. Reporting error is influenced by body weight status and does not occur systematically across different age groups or different dietary survey techniques. It appears that the available methods for assessing the dietary intakes of children are, at best, able to provide unbiased estimates of energy intake only at the group level, while the food intake data of most adolescents are particularly prone to reporting error at both the group and the individual level. Moreover, evidence for the existence of subject-specific responding in dietary assessments challenges the assumption that repeated measurements of dietary intake will eventually obtain valid data. Only limited progress has been made in understanding the variables associated with misreporting in these age groups, the associated biases in estimating nutrient intakes and the most appropriate way to interpret unrepresentative dietary data. Until these issues are better understood, researchers should exercise considerable caution when evaluating all such data. PMID- 15522160 TI - Functional effects of food: what do we know in children? AB - The knowledge base underpinning the setting of nutrient requirements for children and adolescents is not very secure. The advent of the concepts of optimal nutrition and functional foods has encouraged the possible use of functional effects as criteria for adequate nutrition in this age group. Target functions have been identified for growth development and differentiation and for behavioural and cognitive development. However, ideal markers or effects for these are not generally available. Additionally, nutrition in young people should avoid predisposing them to diet-related disease in later life. It is suggested that functional effects should include markers of reduction of risks of disease (nutritional safety) as well of benefits for health and well-being. Such markers of functional effects should be expected to arise from fundamental studies of nutrient-gene interactions and post-genomic metabolism. PMID- 15522161 TI - Effects of diet on behaviour and cognition in children. AB - Diet can affect cognitive ability and behaviour in children and adolescents. Nutrient composition and meal pattern can exert immediate or long-term, beneficial or adverse effects. Beneficial effects mainly result from the correction of poor nutritional status. For example, thiamin treatment reverses aggressiveness in thiamin-deficient adolescents. Deleterious behavioural effects have been suggested; for example, sucrose and additives were once suspected to induce hyperactivity, but these effects have not been confirmed by rigorous investigations. In spite of potent biological mechanisms that protect brain activity from disruption, some cognitive functions appear sensitive to short-term variations of fuel (glucose) availability in certain brain areas. A glucose load, for example, acutely facilitates mental performance, particularly on demanding, long-duration tasks. The mechanism of this often described effect is not entirely clear. One aspect of diet that has elicited much research in young people is the intake/omission of breakfast. This has obvious relevance to school performance. While effects are inconsistent in well-nourished children, breakfast omission deteriorates mental performance in malnourished children. Even intelligence scores can be improved by micronutrient supplementation in children and adolescents with very poor dietary status. Overall, the literature suggests that good regular dietary habits are the best way to ensure optimal mental and behavioural performance at all times. Then, it remains controversial whether additional benefit can be gained from acute dietary manipulations. In contrast, children and adolescents with poor nutritional status are exposed to alterations of mental and/or behavioural functions that can be corrected, to a certain extent, by dietary measures. PMID- 15522162 TI - Nutrition in children and adolescents in Europe: what is the scientific basis? Introduction. PMID- 15522163 TI - Summary report ILSI Europe Workshop: Nutrition in children and adolescents in Europe: what is the scientific basis? PMID- 15522164 TI - Energy and nutrient dietary reference values for children in Europe: methodological approaches and current nutritional recommendations. AB - The Expert Group on the Methodological Approaches and Current Nutritional Recommendations in Children and Adolescents was convened to consider the current situation across Europe with regard to dietary recommendations and reference values for children aged 2-18 years. Information was obtained for twenty-nine of the thirty-nine countries in Europe and a comprehensive compilation was made of the dietary recommendations current up to September 2002. This report presents a review of the concepts of dietary reference values and a comparison of the methodological approaches used in each country. Attention is drawn to the special considerations that are needed for establishing dietary reference values for children and adolescents. Tables are provided of the current dietary reference values for energy and for the macronutrients, vitamins, minerals, trace elements and water. Brief critiques are included to indicate the scientific foundations of the reference values for children and to offer, where possible, an explanation for the wide differences that exist between countries. This compilation demonstrated that there are considerable disparities in the perceived nutritional requirements of European children and adolescents. Although some of this diversity can be attributed to real physiological and environmental differences, most is due to differences in philosophy about the best methodological approach to use and in the way the theoretical approaches are applied. The report highlights the main methodological and technological issues that will need to be resolved before harmonization can be fully considered. Solving these issues may help to improve the quality and consistency of dietary reference values across Europe. However, there are also considerable scientific and political barriers that will need to be overcome and the question of whether harmonization of dietary reference values for children and adolescents is a desirable or achievable goal for Europe requires further consideration. PMID- 15522165 TI - [Association of functional polymorphisms in matrix metalloproteinase-2 (MMP-2) and MMP-9 genes with risk of gastric cancer in a Chinese population]. AB - BACKGROUND & OBJECTIVE: Matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) play important roles in the development of gastric cancer. This study was to investigate the association of functional polymorphisms in the MMP-2 and MMP-9 genes with risk of gastric cancer in a Chinese population. METHODS: MMP-2 1306T/C,and MMP-9 -1562C/T polymorphisms in 228 patients with gastric cancer, and 774 matched healthy controls were detected by polymerase chain reaction (PCR) based denaturing high performance liquid chromatography, and PCR-based restriction fragment length polymorphism analysis. The adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using unconditional logistic regression model. The effect-modified model was used to evaluate the gene-gene interaction. RESULTS: Subjects with the MMP-2 -1306CC genotype had an increased risk of developing gastric cancer compared with those with the MMP-2 -1306TT or CT genotype (adjusted OR, 1.67; 95% CI, 1.17-2.38). No significant association between MMP-9 -1562C/T polymorphism and risk of gastric cancer was observed. However, the polymorphisms in these 2 genes seem to display a gene-gene interaction, with OR of 1.72 (95% CI, 1.07-2.78) for subjects carrying both MMP-2 -1306CC and MMP-9 -1562TT or CT genotypes compared with those carrying both MMP-2 -1306TT or CT and MMP-2 -1306T/C genotypes (likelihood ratio test, P=0.02). CONCLUSION: MMP-2 -1306T/C and MMP-9 -1562C/T polymorphisms may be associated with genetic susceptibility to gastric cancer. PMID- 15522166 TI - [Expression of EDAG-1 gene in human leukemia and lymphoma cell lines]. AB - BACKGROUND & OBJECTIVE: Embryonic development associated gene 1 (EDAG-1), located at chromosome 9q22, is specially expressed in hematopoietic cells, and related to the regulation of hematopoietic system. This study was designed to explore relationship between pathogenesis of leukemia, lymphoma and EDAG-1 through analyzing the structure of EDAG-1 coding region, and its expression in these cell lines. METHODS: Fifteen leukemia and lymphoma cell lines, HEL, K562, HL-60, Namalwa, Raji, J111, Jurkat, HuT 78, MEG-01, U937, 6T-CEM, HPB-ALL, KG-1a, THP-1, and DAMI, were selected to observe the expression of EDAG-1 by reverse transcriptase-polymerase chain reaction (RT-PCR), EDAG-1 cDNA coding fragments (1.5 kb) were purified to construct the corresponding recombinant plasmid. Then, the plasmid was sequenced to analyze mutation of the coding region. The expression of EDAG-1 protein, and mRNA in these cell lines were detected by Western blot, and Northern blot; the rearrangement and amplification of EDAG-1 genome in these cell lines were detected by Southern blot. RESULTS: EDAG-1 mRNA and protein were highly expressed in erythroleukemia cell lines (K-562, HEL), megakaryoblast leukemia cell lines (DAMI, MEG-01), and T cell leukemia cell line (Jurkat), while no gene mutation was found in coding region, no amplification and rearrangement of genome was detected in these cell lines. EDAG-1 was absent in HL 60 cell line, and rearranged in HuT 78 cells. CONCLUSION: EDAG-1 may relate with pathogenesis of erythroleukemia and megakaryoblast leukemia; its coding region may have no relation with the mechanism of its activation. PMID- 15522167 TI - [Promotive effect of decreased cyclin E threshold on cell proliferation]. AB - BACKGROUND & OBJECTIVE: Many studies showed that high expression of Cyclin E promotes cell proliferation, but contrary data was also reported that cell proliferation didn't decrease with low expression of Cyclin E. In addition, we observed that many tumor cells have strong capability of proliferation with low expression of Cyclins, including Cyclin E. This study was to analyze effect of reduced Cyclin E threshold on proliferation of acute lymphocyte leukemia cell line MOLT-4 to explain the above phenomena. METHODS: We have established the model of decreased Cyclin E threshold in MOLT-4 cells by treating cells with low concentration (5 mmol/L) of caffeine for 2, and 4 h. The positive rates of proliferation cell nuclear antigen (PCNA), Ki67, and DNA strand break induction by photolysis (SBIP) were analyzed by flow cytometry. RESULTS: MOLT-4 cells presented sharply decrease of Cyclin E threshold, and increase of positive rates of PCNA, Ki67, and SBIP after treated with caffeine, especially at 2-h point. CONCLUSIONS: Decrease of Cyclin E threshold was accompanied by increase of cell proliferation. MOLT-4 cells may remain high proliferation capability with low level of Cyclin expression. PMID- 15522168 TI - Effects of DNA methylation inhibitor on down-regulation of ribonuclease inhibitor expression in cancer cell lines. AB - BACKGROUND & OBJECTIVE: Human ribonuclease inhibitor (RI) could effectively block angiogenin-induced angiogenesis, and inhibit growth of transplant solid tumors in animals. However, its exact molecular mechanism of antitumor has not been totally ascertained. Many tumor suppressor genes occur loss of expression by aberrant methylation in promoter region. Demethylation, treated with methylation inhibitor 5-aza-2'-deoxycytidine (5-Aza-CdR),can restore the gene expression. To further explore functions of RI, and investigate relationship between RI and tumorigenesis, the study was designed to explore effects of 5-Aza-CdR on expression of RI in cancer cell lines. METHODS: Human breast cancer cell line MCF 7, human gastric cancer cell line BGC-823, human prostate cancer cell line DU 145, and human colon cancer cell line HT-29 were treated with 5-Aza-CdR. Expression of RI was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blot, immunofluorescence, and immunocytochemistry. RESULTS: Expression of RI significantly elevated by 5-Aza-CdR at both mRNA and protein level in MCF-7, BGC-823, and DU-145 cells (P< 0.01). Compared with control cells, mRNA levels of RI in MCF-7, BGC-823, and DU-145 cells treated with 5-Aza-CdR were increased by percentages of 37.2%, 46.0%, and 32.4%, respectively; protein levels of RI were increased by percentages of 26.4%, 20.9%, and 24.4%, respectively; but no obvious change observed in HT-29 cells. CONCLUSION: RI gene may be involved in tumorigenesis of gastric, prostate, and breast cancer. PMID- 15522169 TI - [Inhibitory effect of RNA interference on expression of HPV16 E6 oncogene in cervical cancer cell line CaSki]. AB - BACKGROUND & OBJECTIVE: Tumorigenesis and progression of cervical cancer closely relate with human papilloma virus (HPV) E6 and E7 oncogenes. Ribozyme and antisense oligonucleotides had been used to inhibit the expression of HPV E6 or E7 oncogenes to treat cervical cancer, but problems, including low efficiency, short-period maintenance, hard work, and high costs, still exist. This study was to evaluate the specific inhibitory effect and time-efficiency of RNA interference (RNAi) on HPV16 E6 gene in cervical cancer cell line CaSki. METHODS: The specific small interfering RNA (siRNA) of HPV16 E6 modified by fluorescein was synthesized, and transfected into CaSki cells. The transfection efficiency of siRNA was evaluated by calculating the ratio of fluorescent cells to total cells. Cell apoptosis was evaluated by flow cytometry (FCM). The mRNA level of HPV16 E6 before and after siRNA transfection was measured by RT-PCR, and protein level of HPV16 E6 was measured by Western blot and FCM. RESULTS: The transfection efficiency of siRNA was 81%. Apoptosis rates of CaSki cells at 1, 2, 5, and 9 d after transfection were 7.7%, 11.8%, 37.4%, and 12.6%, respectively. The mRNA level of HPV16 E6 at 1, 2, 5, and 9 d after transfection reduced by 77%, 83%, 59%, and 41%, respectively, but the mRNA level of beta-actin, as internal control, had no change. The inhibition rates of HPV16 E6 protein at 1, 2, 5, and 9 d after transfection were 79.7%, 80.4%, 71.3%, and 57.4%, respectively, but the protein level of Lamin A/C, as internal control, had no change at each time point. CONCLUSION: RNAi exists in CaSki cells, and has specific high efficiency on HPV16 E6 gene. PMID- 15522170 TI - [Inhibitory effect of MT1-MMP antisense nucleotide on invasion of human highly metastatic ovarian cancer cell line SW626]. AB - BACKGROUND & OBJECTIVE: Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) is a newly discovered enzyme, which plays a key role in tumor metastasis. This study was to observe inhibitory effect of MT1-MMP antisense nucleotide on proliferation and invasive potential of human highly metastatic ovarian carcinoma cell line SW626. METHODS: Reverse transcriptase-polymerase chain reaction (RT PCR) was used to amplify MT1-MMP cDNA fragments with 2 different restriction sites at its 5c-end. RT-PCR products were cloned into plasmid pcDNA3.1 in antisense direction. The recombinant pMMP14as was transfected into SW626 cells. Changes of cell proliferation, MT1-MMP protein expression, activities of MMP-2 and MMP-9, and cell invasion ability were detected by MTT assay, Western blot, optimized gelatin zymography, and matrigel in vitro invasion assay, respectively. RESULTS: Antisense MT1-MMP eukaryotic expression vector pMMP14as was constructed successfully. After 48-h transfection with pMMP14as, proliferation of pMMP14as transfected SW626 cells was significantly lower than that of control cells. Compared with control cells, the expression of endogenous MT1-MMP protein in pMMP14as-transfected cells was decreased with a inhibition rate of 65.8%. The activation of proMMP-2 was remarkably inhibited, and the mean invasive cell percentage was (63.3+/-5.8)% in pMMP14as-transfected cells, which was far less than (97.6+/-7.5)% in control cells (P< 0.05). CONCLUSION: Both cell proliferation and invasive potential of SW626 cells were inhibited effectively by antisense MT1-MMP, suggesting that MT1-MMP may be a proper molecular target of anti-invasion therapy for human ovarian cancer. PMID- 15522171 TI - [Inhibitory effect of antisense and dominant-negative AKT2 constructs on proliferation of glioma cell line TJ905]. AB - BACKGROUND & OBJECTIVE: Serine/threonine kinase (AKT) mediates a downstream signal transduction pathway of epidermal growth factor receptor (EGFR), and plays an important role in survival and apoptosis of tumor cells. This study was designed to investigate regulation of antisense AKT2 (AS-AKT2) and dominant negative AKT2 (DN-AKT2) constructs on proliferation and apoptosis of glioma cell line TJ905. METHODS: AS-AKT2 and DN-AKT2 were transfected into TJ905 cells, 3 stably transfected clones were randomly selected from each group, and amplified for further study. The expression of AKT2 was detected by in situ hybridization and Western blot. Proliferation rate of TJ905 cells was evaluated by MTT assay and Ki67 labeling index (Ki67 LI), cell apoptosis was detected by TUNEL method. RESULTS: AKT2 expression was significantly inhibited in TJ905 cells transfected with AS-AKT2, while no significant change observed in TJ905 cells transfected with DN-AKT2. Compared with parental TJ905 cells and TJ905 cells transfected with empty vector, the 6-day survival rates of TJ905 cells transfected with AS-AKT2, and DN-AKT2 dramatically dropped to 49.3%-51.5%, and 50.5%-55.2% (P< 0.001). The Ki67 LI of TJ905 cells transfected with AS-AKT2, and DN-AKT2 dropped to 57.5% 59.3% and 59.2%-61.0%, lower than those of parental TJ905 cells (76.5%), and TJ905 cells transfected with empty vector (74.8%) (P < 0.05). Nearly no apoptotic cells found in parental TJ905 cells or TJ905 cells transfected with empty vector, however, apoptosis indexes (AIs) of TJ905 cells transfected with AS-AKT2, and DN AKT2 prominently increased to 8.8%-8.5%, and 7.5%-7.8% (P< 0.05). The inhibitory effect of AS-AKT2 on cell growth was statistically stronger than that of DN-AKT2 (P< 0.001), while no difference was detected in Ki67 LI and AI between AS-AKT2 and DN-AKT2 transfected TJ905 cells. CONCLUSION: AKT2 pathway may modulate proliferation and apoptosis of glioma cells, indicating that it may be a potential therapeutic target of gene therapy of malignant glioma. PMID- 15522172 TI - [Effects of anti-CXCR4 monoclonal antibody on adhesion and proliferation of human acute myelocytic leukemia cell line HL-60]. AB - BACKGROUND & OBJECTIVE: Stromal cell derived factor-1 (SDF-1), excreted by bone marrow stromal cells, may be involved in shielding of marrow stromal cells from leukemia cells by binding to its receptor CXCR4, but the relation between SDF 1/CXCR4 axis and leukemia cells is unclear. This study was to inhibit activity of SDF-1 by anti-CXCR4 monoclonal antibody 12G5, observe changes of adhesion and proliferation of acute myelocytic leukemia cell line HL-60 co-cultivated with leukemic marrow stromal cells, and to assay potential of inhibiting SDF-1-driven processes on treating marrow residual disease. METHODS: HL-60 cells were co cultured with leukemic marrow stromal cells, and 10 mug/ml 12G5 was added to block SDF-1 activity. Adhesive state, adhesion rate, apoptosis rate, and cell cycle of HL-60 cells were observed after incubation. Living conditions of HL-60 cells were detected by trypan blue exclusion, cell growth curve was protracted. RESULTS: Adhesion rate of 24-h 12G5-incubated HL-60 cells was (39.4+/-7.9)%, while that of control HL-60 cells was (51.4+/-5.9)% (P< 0.05). G0/G1, S, and G2/M phases of 24-h 12G5-incubated HL-60 cells were (55.2+/-4.9)%, (30.4+/-4.1)%, and (14.4+/-5.2)%, respectively, and apoptosis rate was (9.0+/-1.7)%; while those of control HL-60 cells were (44.7+/-2.2)%, (45.3+/-3.7)%, (10.0+/-2.6)%, and (4.0+/ 2.4)%, respectively. T test showed that 12G5 induced more cells entering G0/G1 phase (P< 0.05), and increased cell apoptosis rate(P< 0.05). Compared with control HL-60 cells, survival rate and proliferation of 48-h 12G5-incubated HL-60 cells decreased markedly. CONCLUSION: 12G5 may inhibit adhesion ability and proliferation of HL-60 cells, thus inhibiting SDF-1 activity by 12G5 may be helpful to the treatment of marrow residual disease. PMID- 15522173 TI - [Biological effects of soluble CD40 ligand on lung cancer cell line A549 and its mechanism]. AB - BACKGROUND & OBJECTIVE: Although the roles of CD40 in B cells have been intensively studied, little is known on the function of CD40 in lung cancer cell lines. This study was to investigate biological effects of soluble CD40 ligand (sCD44L) on lung cancer cell line A549 (CD40 positive), and its possible mechanism. METHODS: A549 cells were co-incubated with sCD40L, cell proliferation was detected by MTT assay and 3H-TdR incorporation method. Immunofluorescence technique and flow cytometry (FCM) were used to evaluate changes in cell phenotypes and cell cycle. Cell apoptosis, and expression changes of Bcl-2 and Bax were observed by FCM, reverse transcriptase-polymerase chain reaction (RT PCR), and Western blot. RESULTS: Compared with control cells, proliferation of A549 cells co-incubated with sCD40L was inhibited (P< 0.05). Positive rates of cell surface molecules, CD49e, CD54, TNFRI, and CD95L, in A549 cells co-incubated with sCD40L for 72 h were (61.2+/-4.8)%, (31.2+/-6.1)%,(42.7+/-5.9)%, and (38.2+/ 3.4)%, respectively, while those in control cells were (34.7+/-2.1)%, (7.1+/ 1.6)%, (15.2+/-4.1)%, and (10.1+/-2.3)%, respectively (P< 0.05). However, positive rate of TNFRII in A549 cells co-incubated with sCD40L[(8.7+/-0.8)%] was lower than that in control cells [(58.1+/-3.6)%] (P< 0.05). G1 phase of A549 cells treated with sCD40L for 72 h was (76.0+/-9.1)%, more than that of control cells [(56.7+/-6.9)%], while S phase of sCD40L-treated A549 cells [(10.3+/-5.7)%] was less than that of control cells [(32.7+/-5.5)%]. No significant apoptosis of A549 cells was observed after co-incubated with sCD40L for 72 h, but Bax expression was up-regulated. CONCLUSION: sCD40L may inhibit cell proliferation, cause changes in phenotype and cell cycle of A549 cells, and alter expression of apoptosis-associated gene, such as Bax. PMID- 15522174 TI - [Inhibitory effect of viral macrophage inflammatory protein-II on metastasis of breast cancer cell line MCF-7 through antagonising CXCR4]. AB - BACKGROUND & OBJECTIVE: CXCR4-stromal cell-derived factor-1 (CXCR4-SDF-1alpha) system has been proved to be involved in targeting metastasis of breast cancer. Some antagonists of CXCR4 have inhibitory effects on metastasis of breast cancer. This study was to investigate effect of viral macrophage inflammatory protein-II (vMIP-II), an antagonist of CXCR4, on metastasis of breast cancer cell line MCF 7. METHODS: Proliferation of MCF-7 cells stimulated by vMIP-II of different concentrations (10, 50, 100, 500, and 1 000 ng /ml) was detected by MTT assay, clone formation rate was assessed by agar clone assay. Adhesion and chemotaxis assays were also used to evaluate the effect of vMIP-II on MCF-7 cells in different steps of metastasis. RESULTS: MCF-7 cells treated with vMIP-II of a series of concentrations for 72 h showed no proliferation change (P >0.05). vMIP II (50-1 000 ng /ml) suppressed colony formation of MCF-7 cells in a concentration-dependent manner. After MCF-7 cells treated with 300 ng/ml of vMIP II for different time (0, 0.5, 2, and 6 h), inhibition peak of cell adherence to fibronectin (FN) and Matrigel was observed. The number of migration was low in MCF-7 cells in the presence of vMIP-II of 500 ng/ml (24+/-10) was lower than that of control MCF-7 cells (60+/-9) (P< 0.05). CONCLUSIONS: The clone formation rate of MCF-7 cells may negatively correlates with the concentration of vMIP-II. vMIP II may inhibit MCF-7 cells adhesion to FN and Matrigel, and suppress chemotactic activity of MCF-7 cells toward extracts of human lung protein. PMID- 15522175 TI - [Targeting delivery effect of galactose receptor-mediated c-myc antisense oligonucleotide on human hepatocellular carcinoma cell line Bel-7402]. AB - BACKGROUND & OBJECTIVE: Deliveries of c-myc antisense oligonucleotide (ASODN) mediated by liposome or adenvirus could suppress proliferation of human hepatocellular carcinoma cells, and tumor growth of mice hepatoma model. However, these deliveries lack targeting effects. Receptor-mediated drug delivery is being used in gene and ASODN delivery due to its high targeting efficiency. This study was to evaluate targeting delivery effect of c-myc ASODN mediated by galactose polyethyleneimine (Gal-PEI) on human hepatocellular carcinoma cell line Bel-7402. METHODS: Bel-7402, and lymphoma cell lines, U937 and Raji, were cultured with fluorescence-labeled Gal-PEI-c-myc ASODN, and c-myc ASODN. The uptaking rates of Gal-PEI-c-myc ASODN, and intracellular mean fluorescence intensities of Bel-7402 and U937 cells were tested by flow cytometry. Morphology of Gal-PEI-c-myc ASODN and c-myc ASODN entering Bel-7402, U937, and Raji cells was observed under phase contrast fluorescence microscope. Effects of Gal-PEI-c-myc ASODN of various concentrations on proliferation of these cells were detected by trypan blue dye method. RESULTS: After cultured for 10 min to 4 h, the uptaking rate, and intracellular mean fluorescence intensity of Gal-PEI-c-myc ASODN cultured Bel 7402 cells (88.25%-98.66%, and 38.61%-111.9%) were higher than those of c-myc ASODN cultured Bel-7402 cells, and Gal-PEI-c-myc ASODN cultured U937 cells, significant differences were detected by Poisson test (P < 0.01). Observed by phase-contrast fluorescence microscope, Gal-PEI-c-myc ASODN entered Bel-7402 cells effectively, but can't enter U937, and Raji cells effectively at the same concentration. C-myc ASODN can't enter Bel-7402 cells effectively at the same ASODN concentration. After incubation with Gal-PEI-c-myc ASODN (containing 0.25 1.25 micromol/L of c-myc ASODN) for 48 h, proliferation of Bel-7402 cells was significantly inhibited (P < 0.01=, but no significant differences were detected in U937, and Raji cells (P >0.05). CONCLUSION: Gal-PEI-c-myc ASODN has high targeting delivery effect on Bel-7402 cells, which enhances the intercellular concentration of c-myc ASODN effectively, but it has no such effects on U937, and Raji cells. PMID- 15522176 TI - [Correlation of MUC1 expression to adhesion of breast cancer cell line MDA-MB 231]. AB - BACKGROUND & OBJECTIVE: Recent researches found that an abundant production of mucin protein well correlates with tumor cell metastasis. This study was to investigate inhibitory effect of benzyl-alpha-GalNAc on production of mucin 1 (MUC1), and on adhesion and invasion of breast cancer cell line MDA-MB-231. METHODS: MDA-MB-231 cells were incubated with benzyl-alpha-GalNAc, expression of MUC1 was detected by immunohistochemistry, adhesive ability of MDA-MB-231 cells to artificial basement membrane Matrigel was measured by MTT assay. Gelatin zymography was performed to detect the secretion changes of matrix metalloproteinase-2 (MMP-2) and MMP-9. RESULTS: Compared with control cells, the tumor cells deglycosylated by benzyl-alpha-GalNAc showed lower expression of MUC1 (P < 0.05), and less adhesion to the Matrigel (P< 0.01), the secretion of MMP-2 and MMP-9 suppressed (P< 0.05). CONCLUSION: The expression of MUC1 correlates to adhesion and invasion of MDA-MB-231 cells, benzyl-alpha-GalNAc may weaken adhesion and type IV collagenase-secreting activity of MDA-MB-231 cells by inhibiting MUC1. PMID- 15522177 TI - [Mutation of mitochondrial DNA 4977 bp deletion in laryngeal squamous cell cancer]. AB - BACKGROUND & OBJECTIVE: The 4977 bp deletion of mitochondrial DNA is a molecular marker of cell ageing, and some solid tumors. This study was to detect deletion of mitochondrial DNA 4977 bp in laryngeal squamous cell cancer, and explore its correlation with differentiation of laryngeal cancer tissues. METHODS: Thirty five specimens were collected from patients with pathologically diagnosed laryngeal squamous cell cancer. The total DNA was amplified to identify the 4977 bp deletion of mitochondrial DNA by polymerase chain reaction (PCR), and PCR products were verified by direct sequencing. RESULTS: The 4977 bp deletion detected in 20 of 35 cancer tissues (57.14%), including 5 of 7 with well differentiation, 15 of 24 with moderate differentiation, and 0 of 4 with poor differentiation. Mutation incidence of cancer tissues with well or moderate differentiation were significantly higher than that of cancer tissues with poor differentiation, but there was no significant difference between cancer tissues with well and moderate differentiation. CONCLUSION: The 4977 bp deletion of mitochondrial DNA was detected in laryngeal squamous cell cancer tissues, it might be relates to differentiation of cancer tissues. PMID- 15522178 TI - [Expression of estrogen sulfotransferase (EST) gene and bridging integrator protein-1 (BIN1) gene and their significance in breast tissues]. AB - BACKGROUND & OBJECTIVE: Estrogen sulfotransferase (EST) is an enzyme which metabolizes estrogen into inactive estrogen sulphate. Bridging integrator protein 1 (BIN1) is a novel c-myc-interacting protein with the features of tumor suppressor. EST and BIN1 may be protective in tumorigenesis. This study was to detect the gene expression of EST and BIN1 in breast cancer tissues,and further investigate the carcinogenesis mechanism of breast cancer. METHODS: The mRNA levels of EST and BIN1 in 12 specimens of normal human breast tissue, and 24 specimens of breast cancer were measured by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: EST mRNA and BIN1 mRNA were expressed in all normal human breast tissues, while EST mRNA was decreased in 75.0% (18/24), and lost in 25.0% (6/24) breast cancer specimens; BIN1 mRNA was decreased in 25.0%(16/24), and lost in 66.7.0% (16/24) breast cancer specimens. CONCLUSION: Down-regulation of EST mRNA and BIN1 mRNA may play an important role in the molecular mechanisms of breast carcinogenesis. PMID- 15522179 TI - [Analysis of long-term prognostic factors of epithelial ovarian cancer]. AB - BACKGROUND & OBJECTIVE: Because most of patients with ovarian cancer are in advanced stage when initially noted, and are prone to relapse, ovarian cancer is a tough disease to treat. This study was to determine long-term prognostic factors correlated with prolonged survival of epithelial ovarian cancer patients, give reference for clinical practice. METHODS: Records of 691 patients with epithelial ovarian cancer, admitted in our hospital from Jan. 1986 to Dec. 2000, were retrospectively analyzed. Survival rate was calculated by life table method, and relevant factors were evaluated by univariate analysis. The potential prognostic variables were studied by Cox regression model. RESULTS: The 5-year, and 10-year survival rates of all patients were 56.0%, and 44.6%. Cox regression model revealed that clinical stage, size of tumor residues, pathologic type, histological grade, and number of chemotherapy cycles after surgery emerged as independent prognostic factors. CONCLUSION: Accurate clinical staging and histopathologic assessment, satisfactory cytoreduction, and effective adequate chemotherapy may affect prognosis of ovarian cancer patients. PMID- 15522180 TI - [Surgical treatment of differentiated thyroid carcinoma and analysis of recurrence-related factors]. AB - BACKGROUND & OBJECTIVE: There are many controversies concerning the treatment to patients with differentiated thyroid carcinoma (DTC), focus on appropriate resection extent of DTC, indications for cervical lymph node dissection, et al. This study was to investigate surgical treatment for DTC, and recurrence-related factors. METHODS: Records of 581 patients with DTC treated at Cancer Center, Sun Yat-sen University from Jan. 1st, 1985 to Dec. 31st, 1997 were reviewed retrospectively. Various operation patterns for DTC and their treatment effects, and recurrence-related factors were analyzed. RESULTS: Among 377 patients with DTC who received initial surgery at our hospital, and whose main operation patterns were unilateral lobectomy plus isthmectomy with or without cervical lymph node dissection, 28 (7.43%) recurred, the rate of recurrence in contralateral lobe was 0.80%. Among 195 patients with DTC who received initial surgery in other hospitals, and whose main operation patterns were local mass excision with or without cervical lymph node resection, 143 patients received operation again in our hospital, and 100 of 143 (69.93%) were pathologic positive, positive rate of thyroid was 46.15%, that of cervical lymph node was 48.95%; 47 of 195 patients (24.10%) recurred. Recurrence rate of patients mainly received local mass excision with or without cervical lymph node resection was higher than that of patients mainly received unilateral lobectomy plus isthmectomy with or without cervical lymph node dissection (P >0.05). CONCLUSIONS: Unilateral lobectomy plus isthmectomy with or without cervical lymph node dissection should be recommended as the first treatment for DTC with primary focus localized in unilateral lobe. The initial operation pattern may influence recurrence of DTC. PMID- 15522181 TI - [Microsurgical technique of brain glioma---a report of 183 cases]. AB - BACKGROUND & OBJECTIVE: Prognosis of glioma is still poor, its main treatment is surgery. The extent of tumor resection relates with prognosis. This study was to evaluate the extent of resection, post-operative Karnofsky performance scale (KPS), and survival rate of the glioma patients received microsurgery. METHODS: Records of 183 glioma patients received microneurosurgery were retrospectively analyzed, the extent of resection, post-operative KPS, and survival rate of patients were evaluated. Different microsurgical techniques were applied according to the location of gliomas. En bloc resection was performed for gliomas in non-functional areas by dissecting the tumors along edema area with high-power bipolar electrocoagulation. The tumors in functional areas were separated along cortex sulcus, the central part of tumor was removed firstly, and residual part was resected with low-power electrocoagulation. Gliomas close to important vessels were sucked, and electrocoagulation seldom performed. RESULTS: Among 183 cases of glioma, 85 in non-functional area, 47 in functional area, and 51 close to important vessels. Total and sub-total resection was performed in 163 patients (89.1%). The average post-operative KPS was 74. The KPS was decreased in 23 patients, increased in 44 patients, and stable in 116 patients. Patients were followed up for 12-216 months with an average of 47.8 months. The follow-up rate was 100%. Among 113 patients with long-term follow-up (>/=5 years), 5-year survival rates of low-grade, and high-grade astrocytoma patients were 75.4% (52/69), and 18.2% (8/44). CONCLUSION: Using different microsurgical patterns according to location of glioma, maximal resection of tumor may achieve with protection of neurological function. PMID- 15522182 TI - [Long-term efficacy of radiotherapy on children with nasopharyngeal carcinoma]. AB - BACKGROUND & OBJECTIVE: Nasopharyngeal carcinoma (NPC) in children is a particular type of NPC with poor prognosis. This study was to analyze long-term treatment efficacy, and relevant factors influencing prognosis of NPC in children. METHODS: From Jan. 1975 to Dec. 1995, 47 children,7-14 years old, with NPC diagnosed by pathology received radiotherapy in our hospital. Radiation doses were 52-74 Gy/6-13 weeks [(64.68+/-5.68) Gy] in nasopharynx, and 46-73 Gy/5-13 weeks [(57.77+/-5.86) Gy] in neck; 21 received 1-3 cycles of chemotherapy (cisplatin, bleomycin, 5-fluoroucil, vincristine, and cyclophosphamide) before radiotherapy. Survival rate was calculated by Kaplan-Meier method, and analyzed by log-rank test with SPSS 10.0 software. RESULTS: The 1, 3, 5-year survival rates of 47 patients were 72.3%, 53.2%, and 40.4%, respectively. Clinical stage (P=0.046), mode of biopsy (P=0.024), radiation dose in nasopharynx (P=0.049), and short-term efficacy (P=0.005) correlated with prognosis of these patients. The average height of 15 male children with NPC who survived for more than 5 years was (161.5+/-1.23) cm. Among 3 female children with NPC who survived for more than 5 years, 2 had menstruation disturbance. CONCLUSIONS: Clinical stage, mode of biopsy, radiation dose in nasopharynx, short-term efficacy may influence prognosis of NPC in children. Radiation-induced long-term sequelae of NPC in children should not be overlooked. PMID- 15522183 TI - [Imaging features of Waldeyer's ring in non-Hodgkin's lymphoma and its clinical significance]. AB - BACKGROUND & OBJECTIVE: Waldeyer's ring is the most vulnerable site outside lymph nodes in non-Hodgkin's lymphoma (NHL). Its imaging features in NHL of early stage were similar with those of squamous epithelial cancer, which often leads to misdiagnosis. This study was to discuss imaging features of Waldeyer's ring in NHL, and its clinical significance. METHODS: CT and MRI performances of 149 patients with pathologically diagnosed Waldeyer's ring-NHL were retrospectively analyzed. RESULTS: Among 149 cases of Waldeyer's ring-NHL, 98 (65.8%) were B-cell origin, and 51 (34.2%) were T-cell or NK/T-cell origin; tonsil was the most vulnerable site followed by nasopharyngeal tonsil and other positions. CT and MRI showed that 81 were focal nodules or masses (commonly in B-cell NHL), 36 were diffuse infiltrating growth (commonly in NK/T-cell NHL), 7 were pure ulcer, and 25 were mixed type. The lesions of focal masses are characterized with even CT density and MRI signals. The lesions usually localized in pharynx-mucosa clearance, but rarely in deep space, such as parapharyngeal space, and in skull base. Among 78 cases with involved cervical lymph nodes, 64 were B-cell origin, and 14 were NK/T-cell origin (P< 0.05). CONCLUSION: Multiple positions and centers origin, huge masses, diffuse infiltration growth, and less involvement in deep space and skull-base are typical imaging features of Waldeyer's ring-NHL; CT and MRI may be helpful to diagnose, clinical classifying, and treating Waldeyer's ring-NHL. PMID- 15522184 TI - [MRI performance and diagnosis of meningioma--a report of 126 cases]. AB - BACKGROUND & OBJECTIVE: In MRI study of meningioma, it was lack of large group of patients to evaluate MRI qualitative diagnosis, and no consensus had been achieved concerning problems such as peritumoral edema in meningioma. This study was to summarize main clues for diagnosing meningioma through analyzing MRI performance of 126 patients with meningioma. METHODS: Among 126 patients with meningioma, 32 were syncytial, 35 were fibroblastic, 24 were psammomatous, 9 were angioblastic, 18 were transitional, 3 were papillary, and 5 were malignant. All patients were scanned with T1, T2-weighted imaging (T1WI, T2WI), and contrast enhanced T1WI. RESULTS: Convexity of brain was more likely to be involved, among 126 cases of meningioma, 45 (35.7%) tumors located at convexity of brain. The size of tumor ranged from 1.4 to 9.9 cm. Eighty-one percent of tumors were round or oval in shape. Isointensity or slight hyperintensity of T2WI signals detected in 70.6% patients. The rates of tail sign, and pseudo-capsule were 62.7%, and 49.2%. Extruding sign of brain parenchyma was observed in 83.8% (57/68)of patients with tumor size of > 4 cm. Significantly even and increasing sign in contrast-enhanced T1WI were observed in 104 patients (82.5%). Peritumoral edema occurred in 57 patients (45.1%), and related to tumor size. Other rare signs included cystic changes, bleeding, calcification, osteal changes, and introtumoral vessel symptoms. The correct rate of diagnosis was 95.2%. CONCLUSIONS: MRI performances of meningioma are various. Judgment of extra-brain tumor, typical T2WI signals, tail sign, and significantly even and increasing sign are key factors for diagnosing meningioma. PMID- 15522185 TI - [Diagnostic value of MR diffusion tensor imaging on radiation-induced early brain injury of nasopharyngeal carcinoma after radiotherapy]. AB - BACKGROUND & OBJECTIVE: MR diffusion tensor imaging (DTI), a new MRI technique, can observe water molecule diffusion non-invasively at molecular level. This study was to assess diagnostic value of DTI on post-radiotherapy brain injury by quantitatively analyzing diffuse features of water in bilateral temporal lobes alba of those nasopharyngeal carcinoma (NPC) patients with normal conventional MRI performances. METHODS: DTI was performed in 13 NPC patients with normal conventional MRI performances after radiotherapy, and 21 healthy controls. Isotropic apparent diffusion coefficient (ADCiso) and anisotropic index (AI) were measured in bilateral temporal lobes alba. RESULTS: ADCiso of patient group was (631.30+/-27.83) x 10(-6) mm2/s, while that of control group was (651.76+/-39.10) x 10(-6) mm2/s (P>0.05). Temporal lobes diffusion AI decreased significantly in patient group, mean fractional anisotropy (FA) was 0.405+/-0.042, mean relative anisotropy (RA) was 0.355+/-0.044, and mean 1 minus volume ratio (1-VR) was 0.192+/-0.042; while those in control group were 0.463+/-0.047, 0.418+/-0.052, and 0.257+/-0.055, respectively (P< 0.01). CONCLUSIONS: DTI can early detect subtle alba changes in NPC patients after radiotherapy, which conventional MRI failed to sense. AI is more sensitive than ADCiso, may better describe the diffuse features of water molecules, and evaluate the temporal lobes alba changes before and after NPC radiotherapy. PMID- 15522186 TI - [Correlation of FHIT expression to cell proliferation and metastasis of laryngeal squamous cell carcinoma]. AB - BACKGROUND & OBJECTIVE: Fragile histidine triad (FHIT) gene, a candidate tumor suppressor gene, located at 3pl4.2, spans the most common fragile site, FRA3B, in human genomes. It is altered in many human cancers, such as head and neck squamous cell carcinoma. This study was to investigate correlation of FHIT expression to cell proliferation and metastasis of laryngeal squamous cell carcinoma (LSCC). METHODS: Expression of FHIT protein, and proliferating cell nuclear antigen (PCNA) was detected by SP immunohistochemistry in 41 specimens of LSCC and their matched adjacent non-cancerous epithelium (NCE). RESULTS: Positive rates of FHIT in NCE, and LSCC were 100.0% (41/41), and 46.3% (19/41) (P< 0.01); in LSCC of stage I-II, and stage III-IV were 69.6% (16/23), and 16.7% (3/18) (P< 0.01); in lymph node metastasis group, and no metastasis group were 20.0% (3/15), and 61.5% (16/26) (P< 0.05). PCNA labeling indexes in NCE, and LSCC were (9.98+/ 2.34)%, and (50.71+/-13.64)% (P< 0.01). In LSCC, expression of FHIT negatively correlated with PCNA (P< 0.05). CONCLUSION: Low expression of FHIT might play an important role in cell proliferation and metastasis of LSCC, FHIT might be a useful marker for evaluating biological behaviors of LSCC. PMID- 15522187 TI - [Correlation of expression of heparanase to angiogenesis and prognosis of breast cancer]. AB - BACKGROUND & OBJECTIVE: Heparanase is a heparan sulfate proteoglycan cleaving enzyme. It helps to degrade extracellular matrix and basement membrane, promote angiogenesis, and accelerate tumor metastasis. This study was to investigate correlation of heparanase expression to angiogenesis and prognosis of breast cancer. METHODS: Immunohistochemistry was used to detect heparanase and microvessel density (MVD) in 120 specimens of infiltrative ductal breast cancer, and 10 specimens of normal breast tissue. Correlation of heparanase expression to clinicopathologic factors and prognosis of breast cancer were analyzed using Chi square test, t test, Kaplan-Meier method, and log-rank test. RESULTS: Positive rate of heparanase in breast cancer was 65% (78/120), significantly higher than that in normal breast tissue (0, 0/10) (P< 0.05). MVD in breast cancer was 53.84+/-13.45, significantly higher than that in control group (33.32+/-8.55) (P< 0.01). Expression of heparanase positively correlated with tumor size, histological grade, lymph node metastasis, and clinical stage (P< 0.05) of breast cancer, and negatively correlated with 5-year survival rate (P< 0.05). MVD in heparanase positive group was much higher than that in heparanase negative group (P< 0.05), MVD positively correlated with heparanase expression (r=0.358,P< 0.01). CONCLUSION: Heparanase may promote angiogenesis, and may closely correlate with prognosis of breast cancer. PMID- 15522188 TI - [Predicting cancerization of condyloma acuminatum by testing expression of p16]. AB - BACKGROUND & OBJECTIVE: About 3%-10% of condyloma acuminatum (CA) may develop into cancer. Some studies indicated that homologous deletion of p16 gene is a major factor that causes cancerization of CA. This study was to detect expression of P16 protein in CA tissues and its cancerization tissues, and to investigate relationship of abnormal expression of P16 and cancerization of CA. METHODS: A total of 75 skin biopsy specimens were collected, including 30 normal skin samples (control group), 35 CA samples, and 10 cancerized CA samples. Expression of P16 was tested by LSAB immunohistochemistry, and relationship of P16 and cancerization of CA was statistically analyzed. RESULTS: CA and normal skin tissues showed weakly positive expression of P16, no significant difference exist (P< 0.05). Cancerized CA tissues showed positive or strongly positive expression of P16, significantly stronger than CA and normal skin tissues (P< 0.05). CONCLUSIONS: Positive and strongly positive expression of P16 in CA tissue implied risk of cancerization of CA. P16 may be a useful predictor for cancerization of CA. PMID- 15522189 TI - [CK20 mRNA expression in peripheral blood of patients with gastrointestinal carcinoma and its clinical significance]. AB - BACKGROUND & OBJECTIVE: Detecting expression levels of cytokeratin 19 (CK19), CK20, MUC1, and carcinoembryonic antigen (CEA) in peripheral blood,lymph node,and bone marrow is a major method for diagnosing micro-metastasis in patients with gastrointestinal carcinoma of early stages. This study was to examine the expression, before and after operation, of CK20 mRNA in peripheral blood of patients with gastrointestinal carcinoma, to further explore hematogenous-spread micro-metastasis status of these patients, and significance of CK20 mRNA detection in treatment during perioperation period. METHODS: Peripheral blood samples were collected before, and 2 weeks after operation from 62 patients with gastrointestinal carcinoma,and 22 controls (12 patients with benign gastrointestinal disease, and 10 healthy volunteers). CK20 mRNA expression in the blood samples was specifically detected by nested reverse transcriptase polymerase chain reaction (RT-PCR). Pathologic examination of all cases was done by conventional methods. RESULTS: Of 62 patients with gastrointestinal carcinoma, 34 (54.3%) have positive CK20 mRNA expression before operation,and so did 31 (50.0%) postoperatively, but no CK20 mRNA expressed in 22 controls. In patients of stageI-IV, CK20 mRNA positive rates were 37.5% (3/8), 36.3% (8/22), 66.7% (18/27), and 100% (5/5), respectively, difference between stage I+II (36.9%) and stage III+IV (83.3%) was significant (P< 0.05). Among 35 patients with local lymph node metastasis, 27 (77.1%) were positive for CK20 mRNA; among 27 patients without lymph node metastasis, 7 (25.9%) have positive CK20 mRNA expression (P< 0.01). CK20 mRNA expression was not correlated with tumor cell differentiation,and cancer embolus existance. There was no significant difference in CK20 mRNA expression before and after operation (P >0.05). CONCLUSIONS: CK20 mRNA may be a sensitive marker for detecting micro-metastasis,and recurrence of patients with gastrointestinal carcinoma. It may be helpful for the correct clinical staging,and reasonable treatment for patients. PMID- 15522190 TI - [Improvement and application of methylation-specific polymerase chain reaction]. AB - BACKGROUND & OBJECTIVE: Methylation-specific polymerase chain reaction (MSP) is frequently used to screen DNA methylation state, but it is complicate and time consuming, with high output of non-specific polymerase chain reaction (PCR) products, and diseconomy, these methods need to be improved. RUNX3 gene is a tumor suppressor gene (TSG) in gastric cancer, and inactivated by both hypermethylation and allelic loss. This study was to adopt the improved MSP to detect methylation state of RUNX3 gene in hepatocellular carcinoma (HCC). METHODS: Glassmilk was used to purify and recover DNA modified by sodium of bisulfite; GC-Melt solution was particularly used for amplification of GC-rich sequence to improve PCR specification. Methylation state of RUNX3 gene in 152 specimens of HCC was detected by this improved MSP, conventional MSP was used as control. RESULTS: In conventional MSP, expensive spin columns, and high speed and low temperature centrifuge should be used to purify and recover DNA, taking up at least 16 h in this process. On contrast, in improved MSP, purifying and recovering DNA with glassmilk need no expensive reagent or equipment mentioned above, taking up only about 1 h in the whole process. Compared with conventional MSP, non-specific PCR products of RUNX3 gene were greatly decreased by GC-Melt solution. Hypermethylation of RUNX3 gene detected in 51.9% (79/152) of HCC specimens. CONCLUSION: Improved MSP is a rapid, simple, and effective way to screen DNA methylation. DNA methylation may be an important mechanism of inactivation of RUNX3 gene in HCC. PMID- 15522191 TI - [Application and development of purification in cancer genomics research]. AB - In the study of cancer genomics, a series of purification methods have been invented to collect homogenous cells from heterogeneous cancerated tissue. Tissue microdissection methods, such as manual microdissection, mechanical microdissection, and laser technology, accelerated cancer genomics research. Each method holds both advantage and disadvantage, the selection of methods depends on the demands of study, and the instruments used in study. PMID- 15522192 TI - Correspondence regarding Bharanidharan et al., "Correlations between nucleotide frequencies and amino acid composition in 115 bacterial species". AB - Bharanidharan et al. [Biochem. Biophys. Res. Commun. 315 (2004) 1097-1103] claimed that the frequencies of most amino acids are determined by the dinucleotide composition of the genome. Here, regarding a methodological problem in their work, it is suggested that the standard deviations of amino acid frequencies should be determined to indicate how significant a certain deviation from the predicted frequency is. Furthermore, using a different method that is expected to be more reliable, we suggest that the dinucleotide composition cannot explain the observed frequencies of most amino acids, and the deviations of amino acid frequencies from what dinucleotide composition predicts are larger than to be expected by chance. PMID- 15522193 TI - A key role for heme oxygenase-1 in nitric oxide resistance in murine motor neurons and glia. AB - Nitric oxide is utilized at low levels for intercellular signaling, and at high levels as a cytotoxic weapon during inflammation. Cellular NO resistance can be increased by prior exposure to sublethal NO levels to induce defense gene expression (adaptive NO resistance), which has been correlated with increased expression of heme oxygenase-1 (HO1) and was blocked by a heme oxygenase inhibitor. However, the possibility remained that other activities were affected by the inhibitor. To address this question, we conducted a genetic study of the HO1 role. We show here that primary cultures of spinal motor neurons and glia from homozygous HO1-null mice are strikingly more sensitive to NO cytotoxicity than are cells expressing HO1. Following an exposure to NO, the HO1-deficient cells were much more prone to apoptosis than were HO1-expressing cells with either one or two copies of a functional HO1 gene. These results confirm the in vivo role of HO1 as a front-line defense against NO toxicity in neuronal cells. PMID- 15522194 TI - Interaction of shikimic acid with shikimate kinase. AB - The crystal structure of shikimate kinase from Mycobacterium tuberculosis (MtSK) complexed with MgADP and shikimic acid (shikimate) has been determined at 2.3A resolution, clearly revealing the amino acid residues involved in shikimate binding. In MtSK, the Glu61 strictly conserved in SK forms a hydrogen bond and salt-bridge with Arg58 and assists in positioning the guanidinium group of Arg58 for shikimate binding. The carboxyl group of shikimate interacts with Arg58, Gly81, and Arg136, and hydroxyl groups with Asp34 and Gly80. The crystal structure of MtSK-MgADP-shikimate will provide crucial information for elucidation of the mechanism of SK-catalyzed reaction and for the development of a new generation of drugs against tuberculosis. PMID- 15522195 TI - RNase-L regulates the stability of mitochondrial DNA-encoded mRNAs in mouse embryo fibroblasts. AB - Accelerated decrease in the levels of mitochondrial DNA-encoded mRNA (mt-mRNA) occurs in neuronal cells exposed either to the excitatory amino acid, glutamate or to the sodium ionophore, monensin, suggesting a role of mitochondrial RNase(s) on the stability of mt-mRNAs. Here we report that in mouse embryo fibroblasts that are devoid of the interferon-regulated RNase, RNase-L, the monensin-induced decrease in the half-life of mt-mRNA was reduced. In monensin (250 nM)-treated RNase-L(+/+) cells the average half-life of mt-mRNA, determined after termination of transcription with actinomycin D, was found to be 3h, whereas in monensin treated RNase-L(-/-) cells the half-life of mt-mRNA was >6h. In contrast, the stability of nuclear DNA-encoded beta-actin mRNA was unaffected. Induction of RNase-L expression in mouse 3T3 fibroblasts further decreased the monensin induced reduction in mt-mRNA half-life to 1.5h. The results indicate that the RNase-L-dependent decrease in mtDNA-encoded mRNA transcript levels occurs through a decrease in the half-life of mt-mRNA, and that RNase-L may play a role in the stability of mt-mRNA. PMID- 15522196 TI - A potential pro-angiogenic cell therapy with human placenta-derived mesenchymal cells. AB - Recently several strategies to treat ischemic diseases have been proposed but the ideal way has to be determined. We explored whether human placenta-derived mesenchymal cells (hPDMCs) can be used for this purpose because placenta is very rich in vessels. First, production of human vascular endothelial growth factor (hVEGF) from hPDMCs was examined. The amount of hVEGF secreted by hPDMCs was similar to the amount produced by HeLa cells. hVEGF was barely detected in human umbilical vein endothelial cells (hUVECs) or human peripheral blood mononuclear cells. hVEGF secreted from hPDMCs stimulated the proliferation of hUVECs, indicating its biological activity. Transplantation of hPDMCs to the ischemic limbs of NOD/Shi-scid mice significantly improved the blood flow of the affected limbs. Blood vessel formation was more prominently observed in the limbs of treated mice as compared to the control mice. Real-time RT-PCR revealed that hPDMCs produced hVEGF for at least 7 days after transplantation. Thus, transplantation of hPDMCs could potentially be a promising treatment for human ischemic diseases. PMID- 15522197 TI - Regulation of high glucose-induced apoptosis by mitochondrial NADP+-dependent isocitrate dehydrogenase. AB - A high concentration of glucose has been implicated as a causal factor in initiation and progression of diabetic kidney complications, and there is evidence to suggest that hyperglycemia increases the production of free radicals and oxidant stress. Recently, we demonstrated that the control of mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDPm) to supply NADPH for antioxidant systems. In this report, we demonstrate that modulation of IDPm activity in HEK293 cells, an embryonic kidney cell line, regulates high glucose-induced apoptosis. When we examined the protective role of IDPm against high glucose-induced apoptosis with HEK293 cells transfected with the cDNA for mouse IDPm in sense and antisense orientations, a clear inverse relationship was observed between the amount of IDPm expressed in target cells and their susceptibility to apoptosis. The results suggest that IDPm plays an important protective role in apoptosis of HEK293 cells induced by a high concentration of glucose and may contribute to various pathologies associated with the long-term complications of diabetes. PMID- 15522198 TI - Sex- and tissue-specific expression of maintenance and de novo DNA methyltransferases upon low dose X-irradiation in mice. AB - DNA methylation is crucial for normal development, proliferation, and proper maintenance of genome stability for a given organism. A variety of DNA damaging agents that are known to affect genome stability were also shown to alter DNA methylation patterns. We have recently pioneered the studies in the area of the radiation effects on DNA methylation, and found that radiation exposure led to substantial dose-dependent and tissue-specific DNA hypomethylation, which was much more pronounced in spleen and liver of female animals. The exact mechanisms of radiation-induced DNA hypomethylation are still to be uncovered. We have previously shown that one of those mechanisms may potentially be DNA repair related. Another possible mechanism may be linked to changes in the expression of DNA methyltransferases (DNMTs). In the current study, we examined the radiation induced changes in expression of maintenance DNMT1, and de novo methyltransferases DNMT3a and DNMT3b in spleen and liver of irradiated animals. This was paralleled by the studies of acute and chronic IR-induced methylation changes in spleen and liver of intact animals, as well as in animals with altered sex hormone status. Here we report that radiation-induced DNA methylation changes correlated with radiation-induced alterations in expression of DNA methyltransferases. We present the data on tissue-specificity in radiation induced expression of DNA methyltransferases, and prove that changes in the expression of de novo methyltransferases DNMT3a and DNMT3b are the most important in radiation-induced DNA methylation alterations. We also discuss the role of sex hormones, especially estrogen, in the generation of the sex-specific radiation induced methylation changes. PMID- 15522199 TI - Age-related compensatory activation of pyruvate dehydrogenase complex in rat heart. AB - Mitochondrial uptake and beta-oxidation of long-chain fatty acids are markedly impaired in the aging rat heart. While these alterations would be expected to adversely affect overall pyridine nucleotides, NADH levels do not change significantly with age. This conundrum suggests that specific compensatory mechanisms occur in the aging heart. The comparison of cardiac pyruvate dehydrogenase complex (PDC) kinetics in 4- and 24- to 28-month-old F344 rats revealed a 60% significant increase in V(max) with no change in PDC expression, and a 1.6-fold decrease in the Michaelis constant (K(m)) in old compared to young rats. The observed kinetic adjustments were selective to PDC, as neither the V(max) nor K(m) of citrate synthase changed with age. PDC kinase-4 mRNA levels decreased by 57% in old vs young rat hearts and correlated with a 45% decrease in PDC phosphorylation. We conclude that PDC from old rat hearts catabolizes pyruvate more efficiently due to an adaptive change in phosphorylation. PMID- 15522200 TI - Functional identification of the pro-apoptotic effector domain in human Sox4. AB - Recent studies provide evidence that Sox4 is involved in regulating apoptosis as well as tumorigenesis of various human cancers; however, its role in the apoptotic machinery is not fully understood. Here we describe that the central domain containing glycine-rich region in Sox4, named CD, is a pivotal pro apoptotic domain to induce apoptotic cell death. Deletion of the DNA-binding domain or trans-activation domain in Sox4 did not significantly affect pro apoptotic activity, whereas transient transfection of the high mobility group box or the serine-rich region abrogated the apoptotic activity. Moreover, overexpression of the CD construct (aa 166-342) revealed the apoptotic activity comparable to that of wild-type Sox4, approximately 60% of cell death. Our data suggest that the apoptotic activity of Sox4 can be dissociated from its transcriptional trans-activation and is mediated through its CD. PMID- 15522201 TI - In vitro assembly into virus-like particles is an intrinsic quality of Pichia pastoris derived HCV core protein. AB - Different variants of hepatitis C virus core protein (HCcAg) have proved to self assemble in vitro into virus-like particles (VLPs). However, difficulties in obtaining purified mature HCcAg have limited these studies. In this study, a high degree of monomeric HCcAg purification was accomplished using chromatographic procedures under denaturing conditions. Size exclusion chromatography and sucrose density gradient centrifugation of renatured HCcAg (in the absence of structured RNA) under reducing conditions suggested that it assembled into empty capsids. The electron microscopy analysis of renatured HCcAg showed the presence of spherical VLPs with irregular shapes and an average diameter of 35nm. Data indicated that HCcAg monomers assembled in vitro into VLPs in the absence of structured RNA, suggesting that recombinant HCcAg used in this work contains all the information necessary for the assembly process. However, they also suggest that some cellular factors might be required for the proper in vitro assembly of capsids. PMID- 15522202 TI - Mutations of the POMT1 gene found in patients with Walker-Warburg syndrome lead to a defect of protein O-mannosylation. AB - Walker-Warburg syndrome (WWS) is an autosomal recessive developmental disorder characterized by congenital muscular dystrophy, brain malformation, and structural eye abnormalities. WWS is due to defects in protein O mannosyltransferase 1 (POMT1), which catalyzes the transfer of mannose to protein to form O-mannosyl glycans. POMT1 has been shown to require co-expression of another homologue, POMT2, to have activity. In the present study, mutations in POMT1 genes observed in patients with WWS were duplicated by site-directed mutagenesis. The mutant genes were co-expressed with POMT2 in Sf9 cells and assayed for protein O-mannosyltransferase activity. Expression of all mutant proteins was confirmed by Western blot, but the recombinant proteins did not show any protein O-mannosyltransferase activity. The results indicate that mutations in the POMT1 gene result in a defect of protein O-mannosylation in WWS patients. This may cause failure of binding between alpha-dystroglycan and laminin or other molecules in the extracellular matrix and interrupt normal muscular function and migration of neurons in developing brain. PMID- 15522203 TI - Adrenomedullin induces matrix metalloproteinase-2 activity in rat aortic adventitial fibroblasts. AB - BACKGROUND: The delicate balance of the extracellular matrix (ECM) determines the stiffness of the vascular wall, and adventitial fibroblasts are involved in ECM formation by synthesizing and degrading matrix proteins. In the present study, we examined the effect of the bioactive peptide adrenomedullin (AM) on activity and expression of matrix metalloproteinases (MMPs) in cultured aortic adventitial fibroblasts. METHODS AND RESULTS: In cultured adventitial fibroblasts isolated from aorta of adult Wistar rats, 10(-6)mol/L angiotensin II (Ang II) significantly (p<0.05) down-regulated MMP-2 activity as determined by in vitro gelatin zymography. In contrast, 10(-7)mol/L synthetic rat AM significantly (p<0.05) stimulated zymographic MMP-2 activity by 23%, increasing intracellular cAMP, and AM abolished the action of Ang II, augmenting the MMP-2 activity. Similarly, Ang II down-regulated MMP-2 protein expression assessed by Western blotting, whereas AM increased it. Furthermore, 8-bromo-cAMP, an analogue of cAMP, mimicked the effect of AM, and H-89, an inhibitor for protein kinase A (PKA), significantly decreased the basal and AM-induced MMP-2 activity. CONCLUSION: This study provides a new insight into the biological action of AM and its intracellular signaling system of cAMP/PKA stimulating the matrix degrading enzyme MMP-2, suggesting an important role for this molecule in modulating ECM deposition in the adventitial layer. PMID- 15522204 TI - Characterization of a rhodanese from the cyanogenic bacterium Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa, the rRNA group I type species of genus Pseudomonas, is a Gram-negative, aerobic bacterium responsible for serious infection in humans. P. aeruginosa pathogenicity has been associated with the production of several virulence factors, including cyanide. Here, the biochemical characterization of recombinant P. aeruginosa rhodanese (Pa RhdA), catalyzing the sulfur transfer from thiosulfate to a thiophilic acceptor, e.g., cyanide, is reported. Sequence homology analysis of Pa RhdA predicts the sulfur-transfer reaction to occur through persulfuration of the conserved catalytic Cys230 residue. Accordingly, the titration of active Pa RhdA with cyanide indicates the presence of one extra sulfur bound to the Cys230 Sgamma atom per active enzyme molecule. Values of K(m) for thiosulfate binding to Pa RhdA are 1.0 and 7.4mM at pH 7.3 and 8.6, respectively, and 25 degrees C. However, the value of K(m) for cyanide binding to Pa RhdA (=14 mM, at 25 degrees C) and the value of V(max) (=750 micromol min( 1)mg(-1), at 25 degrees C) for the Pa RhdA-catalyzed sulfur-transfer reaction are essentially pH- and substrate-independent. Therefore, the thiosulfate-dependent Pa RhdA persulfuration is favored at pH 7.3 (i.e., the cytosolic pH of the bacterial cell) rather than pH 8.6 (i.e., the standard pH for rhodanese activity assay). Within this pH range, conformational change(s) occur at the Pa RhdA active site during the catalytic cycle. As a whole, rhodanese may participate in multiple detoxification mechanisms protecting P. aeruginosa from endogenous and environmental cyanide. PMID- 15522205 TI - Toll-like receptor 4 mutation impairs the macrophage TNFalpha response to peptidoglycan. AB - Macrophages produce TNFalpha when infected by bacteria, a response that follows recognition of microbial components by members of the Toll-like receptor (TLR) family. Cells that lack functional TLR4 are known to have markedly diminished responses to Gram-negative lipopolysaccharide. We demonstrate in the present work that peritoneal macrophages derived from strains of mice that carry a spontaneous, inactivating mutation in TLR4 also have impaired production of TNFalpha in response to peptidoglycan, a ligand for TLR2. This impairment is at a step of biosynthesis subsequent to the generation of mRNA. TLR4-activated signals act at this step to enhance peptidoglycan-induced TNFalpha production in wild type mice. Based on these observations, we conclude that macrophages from wild type mice are primed by chronically acting TLR4 signals, probably resulting from exposure to environmental lipopolysaccharide. These signals are required for optimal production of TNFalpha in response to TLR2 stimulation, and are absent in macrophages from TLR4 mutant animals. PMID- 15522206 TI - Abrogation of hepatocyte apoptosis and early appearance of liver dysplasia in ethanol-fed p53-deficient mice. AB - Ethanol consumption represents a major risk factor for cancer development, and a significant fraction of hepatocarcinomas arises in alcoholic liver cirrhosis. Increasing evidence indicates that ethanol acts as a tumor promoter on genetically initiated cells, by increasing the intracellular concentration of reactive oxygen species and promoting tissue necrosis/regeneration and cell proliferation. The tumor suppressor p53 restrains the expansion of carcinogen initiated cells by inducing cell cycle arrest and apoptosis; accordingly, p53 deficient mice develop spontaneous and chemically induced neoplasms at a much higher frequency than normal mice. In normal mice exposed to a subacute (3 weeks) ethanol intoxication, a significant increase in the number of apoptotic hepatocytes was observed in concomitance with the up-regulation of the mitochondrial superoxide scavenger MnSOD, a reliable indicator of oxidative stress. Cell death occurred in the absence of liver inflammation and necrosis. Ethanol-induced hepatocyte apoptosis was completely abrogated in the p53 null background, suggesting that the tumor suppressor is necessary for hepatocyte death by ethanol. Accordingly, p53 -/- MEF were, unlike wild type cells, completely insensitive up to 0.5M ethanol in the culture medium. Strikingly, marked and widespread signs of dysplasia, with nuclear pleomorphisms and initial loss of normal architecture, heralding malignant transformation, were scored in all the mutant mice exposed to ethanol, but not in the control-fed littermates nor in ethanol-fed normal mice. These observations suggest that p53-dependent apoptosis restrains the tumorigenic effect of ethanol on liver cells, in agreement with the frequent loss of p53 function in HCC, and reveal an unexpected carcinogenic potential of alcohol which appears to be independent from the induction of cirrhosis and hepatocyte regeneration. PMID- 15522207 TI - Intracellular glutathione status regulates mouse bone marrow monocyte-derived macrophage differentiation and phagocytic activity. AB - Although a redox shift can regulate the development of cells, including proliferation, differentiation, and survival, the role of the glutathione (GSH) redox status in macrophage differentiation remains unclear. In order to elucidate the role of a redox shift, macrophage-like cells were differentiated from the bone marrow-derived monocytes that were treated with a macrophage colony stimulating factor (M-CSF or CSF-1) for 3 days. The macrophagic cells were characterized by a time-dependent increase in three major symptoms: the number of phagocytic cells, the number of adherent cells, and the mRNA expression of c-fms, a M-CSF receptor that is one of the macrophage-specific markers and mediates development signals. Upon M-CSF-driven macrophage differentiation, the GSH/GSSG ratio was significantly lower on day 1 than that observed on day 0 but was constant on days 1-3. To assess the effect of the GSH-depleted and -repleted status on the differentiation and phagocytosis of the macrophages, GSH depletion by BSO, a specific inhibitor of the de novo GSH synthesis, inhibited the formation of the adherent macrophagic cells by the down-regulation of c-fms, but did not affect the phagocytic activity of the macrophages. To the contrary, GSH repletion by the addition of NAC, which is a GSH precursor, or reduced GSH in media had no effect on macrophage differentiation, and led to a decrease in the phagocytic activity. Furthermore, we observed that there is checkpoint that is capable of releasing from the inhibition of the formation of the adherent macrophagic cells according to GSH depletion by BSO. Summarizing, these results indicate that the intracellular GSH status plays an important role in the differentiation and phagocytosis of macrophages. PMID- 15522208 TI - Transcriptional control of cystine/glutamate transporter gene by amino acid deprivation. AB - Recent studies have demonstrated that depletion of amino acids results in the induction of several genes and that a genomic cis-element termed amino acid response element (AARE) is required for the induction. System x(c)(-) is an anionic amino acid transport system highly specific for cystine and glutamate, and its activity is known to be induced by cystine deprivation. This transporter is composed of two protein components, xCT and 4F2 heavy chain, and xCT is thought to mediate the transport activity. In the present study, the molecular mechanism for the induction of xCT by amino acid deprivation has been investigated. In mouse NIH3T3 cells, the activity of system x(c)(-) and xCT mRNA is induced not only by deprivation of cystine but also by deprivation of other amino acids. Two AAREs, each located in the opposite direction with an intervening sequence, were found in the 5'-flanking region of the mouse xCT gene. Promoter analysis revealed that both AAREs were necessary for the maximal induction of xCT mRNA in response to the amino acid deprivation. Glucose deprivation had no effect on the induction of the activity of system x(c)(-). Electrophoretic mobility shift assay showed that ATF4, but not ATF2, is involved in the amino acid control of xCT expression. These results demonstrate that xCT is a new member of the proteins whose transcriptional control by the amino acid deprivation is mediated by AARE. PMID- 15522209 TI - Differential regulation of phospholipase Cgamma subtypes through FcepsilonRI, high affinity IgE receptor. AB - The high affinity IgE receptor (FcepsilonRI) usually exists as a tetramer composed of alphabetagamma2 subunits. The COOH-tail of beta and gamma subunits contains consensus sequence termed 'immunoreceptor tyrosine-based activation motif' (ITAM). Tyrosine phosphorylated ITAM interacts with signaling proteins that contain the Src homology domain, forming a main amplifying and signaling route for FcepsilonRI. Unlike the COOH-tail, the functional role of NH(2)-tail of beta subunit in the signaling of FcepsilonRI is not clear because it lacks the ITAM sequences. To study the roles of NH(2)-tail of beta subunit, the cDNA library of RBL-2H3 cells was screened by yeast two-hybrid assay, and the NH(2) tail of the beta subunit was found to interact with phospholipase Cgamma2 (PLCgamma2) but not with PLCgamma1. Since both PLCgamma1 and PLCgamma2 are expressed in RBL-2H3 cells and they possess identical cellular functions, the functional meaning of the protein-protein interaction between PLCgamma2 and NH(2) tail of beta subunit was studied by comparing the regulatory pathways that control the FcepsilonRI-mediated tyrosine phosphorylation of the two enzymes. Our study shows that PI3-kinase and PMA-sensitive PKCs were required exclusively for the FcepsilonRI-mediated tyrosine phosphorylation of PLCgamma1. Also the FcepsilonRI-mediated tyrosine phosphorylation of PLCgamma1 was more sensitive to the inhibitors of Src and Syk kinases. These results therefore suggest that PLCgamma1 is involved in dynamic regulation of protein kinase C activity and inositol triphosphate levels in response to cellular needs. In contrast, PLCgamma2, through continuous interaction with the NH(2)-tail of beta subunit, co localizes with FcepsilonRI in the same signaling domain, and maintains the basal cellular PLC activity. PMID- 15522210 TI - Genomewide analysis of gene expression associated with Tcof1 in mouse neuroblastoma. AB - Mutations in the Treacher Collins syndrome gene, TCOF1, cause a disorder of craniofacial development. We manipulated the levels of Tcof1 and its protein treacle in a murine neuroblastoma cell line to identify downstream changes in gene expression using a microarray platform. We identified a set of genes that have similar expression with Tcof1 as well as a set of genes that are negatively correlated with Tcof1 expression. We also showed that the level of Tcof1 and treacle expression is downregulated during differentiation of neuroblastoma cells into neuronal cells. Inhibition of Tcof1 expression by siRNA induced morphological changes in neuroblastoma cells that mimic differentiation. Thus, expression of Tcof1 and treacle synthesis play an important role in the proliferation of neuroblastoma cells and we have identified genes that may be important in this pathway. PMID- 15522211 TI - PKCtheta is required for the activation of human T lymphocytes induced by CD43 engagement. AB - The turnover of phosphoinositides leading to PKC activation constitutes one of the principal axes of intracellular signaling. In T lymphocytes, the enhanced and prolonged PKC activation resulting from the engagement of the TcR and co-receptor molecules ensures a productive T cell response. The CD43 co-receptor promotes activation and proliferation, by inducing IL-2 secretion and CD69 expression. CD43 engagement has been shown to promote phosphoinositide turnover and DAG production. Moreover, PKC activation was found to be required for the activation of the MAP kinase pathway in response to CD43 ligation. Here we show that CD43 engagement led to the membrane translocation and enzymatic activity of specific PKC isoenzymes: cPKC (alpha/beta), nPKC (epsilon and theta;), aPKC (zeta) and PKCmu. We also show that activation of PKCtheta; resulting from CD43 ligation induced CD69 expression through an ERK-dependent pathway leading to AP-1, NF kappaB activation and an ERK independent pathway promoting NFAT activation. Together, these data suggest that PKCtheta; plays a critical role in the co stimulatory functions of CD43 in human T cells. PMID- 15522212 TI - Association of VEGF genotype with mRNA level in colorectal adenocarcinomas. AB - The mRNA expression of vascular endothelial growth factor (VEGF) was evaluated in colorectal adenocarcinomas and adjacent noncancerous colorectal tissues in 18 Japanese patients. The expression was confirmed to be up-regulated in the colorectal adenocarcinomas, when compared with the noncancerous tissues. Twelve genotypes of VEGF: six positions in the promoter region, two in the 5'UTR, and four in the 3'UTR, and their association with the expression of VEGF mRNA were evaluated. While G-1877A, T-1455C, G-1154A, C702T, and G1612A were not detected, C-2578A, T-1498C, G-1190A, C-634G, C-7T, C936T, and C1451T were found at allele frequencies of 4/36, 15/36, 15/36, 20/36, 8/36, 6/36, and 6/36, respectively, suggesting that C-2578A, G-1154A, and G1612A were associated with a decreased risk for colorectal adenocarcinoma. T-1498C (G-1190A) and C-7T were found to be associated with higher levels of VEGF mRNA, and may be a risk factor for the development of liver metastasis and/or prognosis of colorectal adenocarcinoma. PMID- 15522213 TI - Advanced glycation end products-modified proteins and oxidized LDL mediate down regulation of leptin in mouse adipocytes via CD36. AB - Advanced glycation end products (AGE)-modified proteins as well as oxidized-LDL (Ox-LDL) undergo receptor-mediated endocytosis by CHO cells overexpressing CD36, a member of class B scavenger receptor family. The purpose of the present study was to examine the effects of glycolaldehyde-modified BSA (GA-BSA) as an AGE ligand and Ox-LDL on leptin expression in adipocytes. GA-BSA decreased leptin expression at both protein and mRNA levels in 3T3-L1 adipocytes and mouse epididymal adipocytes. Ox-LDL showed a similar inhibitory effect on leptin expression in 3T3-L1 adipocytes, which effect was protected by N-acetylcysteine, a reactive oxygen species (ROS) inhibitor. Binding of (125)I-GA-BSA or (125)I-Ox LDL to 3T3-L1 adipocytes and subsequent endocytic degradation were inhibited by a neutralizing anti-CD36 antibody. Furthermore, this antibody also suppressed Ox LDL-induced leptin down-regulation. These results clarify that the interaction of GA-BSA and Ox-LDL with CD36 leads to down-regulation of leptin expression via ROS system(s) in 3T3-L1 adipocytes, suggesting that a potential link of AGE- and/or Ox-LDL-induced leptin down-regulation might be linked to insulin-sensitivity in metabolic syndrome. PMID- 15522214 TI - Dynamic iodide trapping by tumor cells expressing the thyroidal sodium iodide symporter. AB - The thyroidal sodium iodide symporter (NIS) in combination with various radioactive isotopes has shown promise as a therapeutic gene in various tumor models. Therapy depends on adequate retention of the isotope in the tumor. We hypothesized that in the absence of iodide organification, isotope trapping is a dynamic process either due to slow efflux or re-uptake of the isotope by cells expressing NIS. Iodide efflux is slower in ARH-77 and K-562 cells expressing NIS compared to a thyroid cell line. Isotope retention half times varied linearly with the number of cells expressing NIS. With sufficient NIS expression, iodide efflux is a zero-order process. Efflux kinetics in the presence or absence of perchlorate also supports the hypothesis that iodide re-uptake occurs and contributes to the retention of the isotope in tumor cells. Iodide organification was insignificant. In vivo studies in tumors composed of mixed cell populations confirmed these observations. PMID- 15522215 TI - First report of a glutamine-rich antifungal peptide with immunomodulatory and antiproliferative activities from family Amaryllidaceae. AB - This represents the first report of purification of a glutamine-rich antifungal peptide from family Amarylliaceace. The peptide, designated as nartazin, was purified from the bulbs of the Chinese daffodil Narcissus tazetta var. chinensis by means of ion-exchange chromatography and affinity chromatography. Its molecular mass was 7.1kDa, as determined by SDS-PAGE and gel filtration. Nartazin stimulated proliferation of mouse splenocytes and bone marrow cells but inhibited proliferation of leukemia L1210 cells. It also inhibited translation in a cell free rabbit reticulocyte lysate system. The sequence of its first 20 N-terminal residues was characterized by an abundance of glutamine. The peptide possessed antifungal activity on four phytopathogenic fungi. Its activity was retained after incubation with bovine trypsin and chymotrypsin (enzyme: substrate ratio 1:10 w/w) at 37 degrees C for 1h but was attenuated after treatment with proteinase K. The data revealed its pronounced resistance to proteolytic digestion. PMID- 15522216 TI - Studying protein isoforms of the adaptor SETA/CIN85/Ruk with monoclonal antibodies. AB - SETA/CIN85/Ruk is a multifunctional adaptor protein involved in signal transduction and attenuation downstream of receptor tyrosine kinases. It has a modular structure, and various isoforms that combine different protein-protein interaction domains have been proposed based on cDNA analysis. As a first step towards understanding SETA/CIN85/Ruk isoforms at the protein level, we have characterized 5 monoclonal antibodies against this protein. Three of these were used to study lysates fractionated on a pH gradient, leading to the identification of various SETA/CIN85/Ruk proteins on the basis of pI and apparent molecular weight. While good correspondence with proteins predicted from cDNA analysis was found for two isoforms, in most cases it was not possible to make an unequivocal assignment. We conclude that additional splice variants remain to be described, and that a deeper understanding of SETA/CIN85/Ruk post-translational processing and modification is necessary to gain further understanding of this complex gene product. PMID- 15522217 TI - Mutational analysis of the interaction between insulin receptor and IGF-I receptor with c-Crk and Crk-L in a yeast two-hybrid system. AB - The SH2/SH3 adapter proteins of the Crk family are potent signal transducers after receptor tyrosine kinase stimulation with insulin or IGF-1. We have employed a yeast two-hybrid approach and mutational analysis to dissect the capabilities of the insulin receptor and the IGF-I receptor to directly associate with Crk isoforms. Insulin receptor stably recruits full length Crk by association with its SH2 domain in an auto-phosphorylation dependent manner. In contrast, interaction of the IGF-I receptor with the Crk-IISH2 domain was only detectable when Crk-II was truncated in its C-terminal part, indicating the transient nature of this interaction. From these data it can be concluded that members of the insulin receptor family activate Crk proteins in a differential manner. PMID- 15522218 TI - LPS-mediated NFkappaB activation varies between activated human hepatic stellate cells from different donors. AB - The activation of hepatic stellate cells (HSC) is recognized as the key event of hepatic fibrosis [Virchows Arch. 430 (1997) 195; Semin. Liver Dis. 21 (2001) 437; Front. Biosci. 7 (2002) d808]. NFkappaB has been associated with the development of the activated phenotype, the expression of proinflammatory genes, and with promoting survival of activated HSC. High levels of circulating endotoxin are observed in liver fibrosis and several lines of evidence indicate that LPS plays an important role in chronic liver disease. Here, we investigated the LPS-induced NFkappaB activation in activated HSC from different human donors. HSC were isolated from liver specimens obtained during surgical liver resection and were activated by culturing on plastic. LPS-induced NFkappaB activity and IL-8 expression revealed a significant correlation but differed significantly comparing HSC from individual donors. These variations seen in LPS mediated NFkappaB activation and chemokine secretion between HSC from different donors in vitro may contribute to differences seen in vivo between patients in the progression of fibrosis and the degree of inflammation during chronic liver disease. PMID- 15522219 TI - The developing reproductive 'sink' induces oxidative stress to mediate nitrogen mobilization during monocarpic senescence in wheat. AB - Removal of reproductive 'sink,' i.e., spikelets from wheat, after anthesis delays the rate of flag leaf senescence. Oxidative stress and the oxidative damage to proteins were studied in relation to nitrogen mobilization in wheat plants showing normal and delayed senescence. Wheat plants lacking a reproductive sink showed decreased oxidative stress, lower lipid peroxidation and maintained higher protein, oxidatively damaged proteins, and nitrogen levels as compared to plants with reproductive sink during monocarpic senescence. Oxidative damage to the proteins when not followed by high proteolytic activities led to a slower nitrogen mobilization in wheat plants lacking a reproductive sink. Thus, the influence of the reproductive sink was due to its ability to drive forward the nitrogen mobilization process through high ROS levels which mediated both damage to the proteins and influenced proteolytic activities. PMID- 15522220 TI - Cellular signaling mediated by calphoglin-induced activation of IPP and PGM. AB - Universal protein networks conserved from bacteria to animals dictate the core functions of cells. Inorganic pyrophosphatase (IPP) is an essential enzyme that plays a pivotal role in a broad spectrum of cellular biosynthetic reactions such as amino acid, nucleotide, polysaccharide, and fatty acid biosynthesis. However, the in vivo cellular regulation mechanisms of IPP and another key metabolic enzyme, phosphoglucomutase (PGM), remain unknown. This study aimed to examine the universal protein regulatory network by utilizing genome sequences, yeast proteomic data, and phosphoryl-transfer experiments. Here we report a novel human protein, henceforth referred to as calphoglin, which interacts with IPP and activates it. Calphoglin enhances PGM activity through the activated IPP and more directly on its own. Protein structure and assembly, catalytic function, and ubiquitous cellular localization of the calphoglin (-IPP-PGM) complex were conserved among Escherichia coli, yeast, and mammals. In the rat brain, calphoglin mRNA was enriched in the hippocampus and the cerebellum. Further, the linkage of the calphoglin complex to calcium signaling was demonstrated by its interactive co-localization within the calmodulin/calcineurin signaling complex, by Ca(2+)-binding and Ca(2+)-controlled activity of calphoglin-IPP, and by calphoglin-induced enhancement of microsomal Ca(2+) uptake. Collectively, these results suggest that the calphoglin complex is a common mechanism utilized in mediating bacterial cell metabolism and Ca(2+)/calmodulin/calcineurin-dependent mammalian cell activation. This is the first report of an activator of IPP and PGM, a function novel to proteins. PMID- 15522221 TI - Molecular cloning, sequence analysis, and expression of a cDNA encoding the luciferase from the glow-worm, Lampyris turkestanicus. AB - The first cDNA from lampyridae encoding a glow-worm luciferase from lantern mRNA of Lampyris turkestanicus has been cloned, sequenced, the amino acid sequence predicted, and the sequence reported to GenBank. The cDNA was 1644 base pairs in length and coding a 547-residue protein. The deduced amino acid sequence of the luciferase gene of L. turkestanicus showed 98.7% and 95.8% identity to Lampyris noctiluca and Pyrocoelia rufa, respectively. Phylogenetic analysis further confirmed that the deduced amino acid sequences of L. turkestanicus luciferase gene belong to the same subfamily, Lampyrinae. The cDNA encoding the luciferase of L. turkestanicus was expressed as a 62kDa band in recombinant Escherichia coli and showed green luminescence in the presence of luciferin. Amongst amino acid differences of L. turkestanicus and L. noctiluca (its clade) there are two important substitutions. Signature amino-acid sequences and motifs found in the deduced sequence are CK2-phospho site, ASN-glycosylation, myristoylation site, PKC-phospho site, microbodies C-terminal targeting signal, and AMP-binding domain. PMID- 15522222 TI - Inhibitory mechanism of chroman compound on LPS-induced nitric oxide production and nuclear factor-kappaB activation. AB - 6-Hydroxy-7-methoxychroman-2-carboxylic acid phenylamide (KL-1156) is a novel chemically synthetic compound. In the present study, the chroman KL-1156 compound was found to inhibit lipopolysaccharide (LPS)-induced nitric oxide production in macrophages RAW 264.7. KL-1156 compound attenuated LPS-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), in parallel, and inhibited LPS-induced iNOS promoter activity, indicating that the chroman compound down-regulated iNOS expression at transcription level. As a mechanism of the anti-inflammatory action shown by KL-1156 compound, suppression of nuclear factor (NF)-kappaB has been documented. KL-1156 compound exhibited a dose dependent inhibitory effect on LPS-induced NF-kappaB transcriptional activity in macrophages RAW 264.7. Furthermore, the compound inhibited LPS-induced nuclear translocation of NF-kappaB p65 and DNA binding activity of NF-kappaB complex, in parallel, but did not affect IkappaBalpha degradation. Taken together, this study demonstrated that chroman KL-1156 compound interfered with nuclear translocation step of NF-kappaB p65, which was attributable to its anti-inflammatory action. PMID- 15522224 TI - The metabolism of beta-amyloid converting enzyme and beta-amyloid precursor protein processing. AB - Herein we investigated the processing of beta-secretase (BACE), implicated in Alzheimer's disease through processing of beta-amyloid precursor protein (betaAPP), into smaller metabolites. Four products of approximately 34, approximately 12, approximately 8, and approximately 5kDa were identified, none of which were generated autocatalytically. The approximately 34 and approximately 12kDa forms are held together by disulfide bridges. The approximately 34kDa form results from two cleavages: an N-terminal processing at RLPR(45) downward arrow by furin/PC5, and a C-terminal cleavage at SQDD(379) downward arrow by an unknown enzyme that also releases the C-terminal approximately 12kDa product. Microsequencing of the approximately 8 and approximately 5kDa fragments showed that they are the result of processing at VVFD(407) downward arrow and DMED(442) downward arrow, respectively. Mutagenesis of the identified cleavage sites revealed that the mutants D379A, D379L or D379E block the degradation of BACE into the approximately 12kDa product, confirming the importance of Asp(379). Notably, the D379E mutant results in higher betaAPP derived C99 levels. In contrast, D442A or D442E did not affect the production of the approximately 8 or approximately 5kDa products. The levels of the approximately 8 and approximately 5kDa products are significantly lower in the mutant D407A but less so D407E, likely due to the low efficacy of ER exit of the D407A mutant. Indeed, while co expression of betaAPP with BACE results in enhanced production of Abeta(11-40), the D407A mutant produces mostly Abeta(40). PMID- 15522223 TI - Dimer formation of receptor activator of nuclear factor kappaB induces incomplete osteoclast formation. AB - Receptor activator of nuclear factor kappaB-ligand (RANKL) transduces a differentiation signal appropriate to osteoclasts likely through induction a receptor homotrimer; however, biological importance of RANK-trimerizarion is unknown. To address the signaling mechanism of the RANK receptor, we analyzed the effect of two different types of homodimer inducers RANK-TM-FKBP36v and hEpoR RANK-TM on osteoclastogenesis. Dimerizing component FKBP36v or extracellular portion of human erythropoietin receptor (hEpoR) was fused to RANK lacking the extracellular domain, and the dimerization of this fusion protein was induced by addition of the chemical inducer of dimerization AP20187 or erythropoietin, respectively. Such treatment resulted in induction of TRAP-activity, a marker of osteoclast in a dose dependent manner, with an efficiency equivalent to that of induction by RANKL. However, dimerized-RANK-induced osteoclasts showed relatively low levels of multinucleation, pit forming activity, and expression of calcitonin receptor and cathepsin K, compared with osteoclasts which were induced in the presence of RANKL. As expression of nuclear factor of activated T cells 1 (NFATc1) was also reduced in dimerized-RANK-induced osteoclasts, RANK oligomerization by RANKL is a critical event to generate fully matured osteoclasts through upregulation of NFATc1. PMID- 15522225 TI - Validating siRNA using a reporter made from synthetic DNA oligonucleotides. AB - Only a small fraction of all siRNAs are effective in silencing their target genes, and siRNA efficacy can only be determined experimentally. Previously described reporter-based siRNA validation methods all rely on the availability of physical cDNA clones, and this limits the high throughput applicability of the method. In the current report, we used short synthetic DNA fragment containing a siRNA targeting site, instead of cDNA, to fuse with a reporter gene. When targeting such transcripts with different siRNAs, we found that such constructs can faithfully report the efficacy of the corresponding siRNAs in a sequence specific manner, even when the inserted DNA fragment is essentially only long enough to cover the targeting site. The efficacy of both vector-based siRNA and synthetic siRNA can be evaluated using this system. Since only readily available short synthetic DNA fragments are needed for forming the evaluation vector, this method provides an appealing way of validating siRNAs in high throughput. PMID- 15522226 TI - Endoplasmic reticulum stress and N-glycosylation modulate expression of WFS1 protein. AB - Mutations of the WFS1 gene are responsible for two hereditary diseases, Wolfram syndrome and low frequency sensorineural hearing loss. The WFS1 protein is a glycoprotein located in the endoplasmic reticulum (ER) membrane but its function is poorly understood. Herein we show WFS1 mRNA and protein levels in pancreatic islets to be increased with ER-stress inducers, thapsigargin and dithiothreitol. Another ER-stress inducer, the N-glycosylation inhibitor tunicamycin, also raised WFS1 mRNA but not protein levels. Site-directed mutagenesis showed both Asn-663 and Asn-748 to be N-glycosylated in mouse WFS1 protein. The glycosylation defective WFS1 protein, in which Asn-663 and Asn-748 had been substituted with aspartate, exhibited an increased protein turnover rate. Consistent with this, the WFS1 protein was more rapidly degraded in the presence of tunicamycin. These data indicate that ER-stress and N-glycosylation play important roles in WFS1 expression and stability, and also suggest regulatory roles for this protein in ER-stress induced cell death. PMID- 15522227 TI - Polo-box motif targets a centrosome regulator, RanGTPase. AB - Mammalian polo-like kinase (Plk) acts at various stages in early and late mitosis. Plk1 localizes in the centrosome, the central spindle, the midbody as well as the kinetochore. The non-catalytic region in the C-terminus of Plk1 has conserved sequence motifs, named polo-boxes. These motifs are important for Plk localization. GFP protein fused with the core sequences of polo-box (50 amino acids) localized Plk to target organelles. We screened for Plk interacting proteins by constructing a tandem repeat of the polo-box motif, and used it as bait in the two-hybrid system with HeLa cell cDNA library. RanGTPase was detected as a positive clone. Through in vitro and in vivo protein binding analysis in synchronized cells by thymidine block and by nocodazole treatment, we confirmed the interaction between endogenous Ran and Plk1. We showed that endogenous Ran and Plk1 proteins were co-localized to centrosomes, which is a major target organelle of endogenous Plk1, in early mitotic cells by immunofluorescence. Finally, we demonstrated that Plk1 phosphorylated RanBPM, a Ran-binding protein in microtubule organizing center, through the interaction with Ran. These data suggested that the core motif of polo-box is sufficient for Plk1-targeting, and that Plk1 may play roles in centrosome through recruitment and/or activation of Ran/RanBPM proteins. PMID- 15522228 TI - Global analysis of the expression patterns of transcriptional regulatory factors in formation of embryoid bodies using sensitive oligonucleotide microarray systems. AB - We manufactured a highly sensitive oligonucleotide microarray system comprised entirely of transcription regulatory factors (a TF oligo microarray) in order to comprehensively analyze the expression profiles of transcription factors in mice. We compared the expression profiles of transcription regulatory factors in mouse embryonic stem (ES) cells and ES-differentiated cells by using this TF oligo microarray, a cDNA microarray, a GeneChip system, and quantitative RT-PCR. The TF oligo microarray was able to comprehensively analyze the expression profile of transcription regulatory factors. In addition, we used the manufactured TF oligo microarray to analyze the expression patterns of transcriptional regulatory factors during the formation of embryoid bodies. The TF array was able to reveal the chronologic expression profile of transcription regulatory factors involved in embryogenesis or the maintenance of pluripotency in ES cells. PMID- 15522229 TI - The NC1 domain of human collagen IV is necessary to initiate triple helix formation. AB - Type IV collagen is a heterotrimeric molecule, which contains the N-terminal 7S, a central triple-helical domain, and the globular C-terminal NC1 domain. A zipper like mechanism of triple helix formation, starting from the C-terminus, has been proposed for most collagens but for collagen type IV there has only been indirect evidence so far. In this study we expressed trimeric human collagen type IV to compare the effects of different structural variants on the formation of collagen IV molecules. Our data show that the NC1 but not 7S domain is essential for the chain association and initiation of triple helix formation. This strongly suggests an N-to-C terminal mechanism of triple helix formation. Additionally, we could show that the human alpha2(IV) chain can form chimeric alpha1.alpha1.alpha2(IV) heterotrimers with mouse subunits when expressed in PF HR9 cells. PMID- 15522230 TI - NMR structure of the glucose-dependent insulinotropic polypeptide fragment, GIP(1 30)amide. AB - Glucose-dependent insulinotropic polypeptide is an incretin hormone that stimulates insulin secretion and reduces postprandial glycaemic excursions. The glucose-dependent action of GIP on pancreatic beta-cells has attracted attention towards its exploitation as a potential drug for type 2 diabetes. Use of NMR or X ray crystallography is vital to determine the three-dimensional structure of the peptide. Therefore, to understand the basic structural requirements for the biological activity of GIP, the solution structure of the major biologically active fragment, GIP(1-30)amide, was investigated by proton NMR spectroscopy and molecular modelling. The structure is characterised by a full length alpha helical conformation between residues F(6) and A(28). This structural information could play an important role in the design of therapeutic agents based upon GIP receptor agonists. PMID- 15522231 TI - Inhibition of endothelial cell functions by novel potential cancer chemopreventive agents. AB - Endothelial cells (EC) play a major role in tumor-induced neovascularization and bridge the gap between a microtumor and growth factors such as nutrients and oxygen supply required for expansion. Immortalized human microvascular endothelial cells (HMEC-1) were utilized to assess anti-endothelial effects of 10 novel potential cancer chemopreventive compounds from various sources that we have investigated previously in a human in vitro anti-angiogenic assay. These include the monoacylphloroglucinol isoaspidinol B, 1,2,5,7-tetrahydroxy anthraquinone, peracetylated carnosic acid (PCA), isoxanthohumol, 2,2',4' trimethoxychalcone, 3'-bromo-2,4-dimethoxychalcone as well as four synthetic derivatives of lunularic acid, a bibenzyl found in mosses [Int. J. Cancer Prev. 1 (2004) 47]. EC proliferation was inhibited with half-maximal inhibitory concentrations from 0.3 to 49.6muM, whereas EC migration was affected by most compounds at sub-micromolar concentrations. PCA and the bibenzyl derivative EC 1004 potently prevented differentiation of HMEC-1 into tubule-like structures. Overall, our data indicate that inhibition of endothelial cell function contributes to various extents to the chemopreventive or anti-angiogenic potential of these lead compounds. PMID- 15522232 TI - Myosin heavy chain isoform mRNA and protein levels after long-term paralysis. AB - To assess the long-term influence of paralysis on muscle phenotypic mRNA and protein expression, the effects of spinal cord transection (ST) on myosin heavy chain (MyHC) isoform mRNA and protein levels in the soleus and medial gastrocnemius (MG) muscles of rats were analyzed. Control soleus contained predominantly MyHC-I with low amounts of MyHC-IIa and IIx mRNAs. After ST, MyHC-I mRNA decreased to approximately 15%, MyHC-IIa was increased by 75-200%, and MyHC IIx was elevated by 8-10x. Low level expression of MyHC-IIb was observed post-ST, suggesting that reduced activity is not a primary stimulus for MyHC-IIb expression. Adaptations in mRNA preceded protein adaptations in the soleus. Although MyHC-I protein in the MG was reduced post-ST, no other consistent changes occurred. The relative lack of adaptation to ST by the MG suggests that the reduced activity and load bearing encountered by the MG were insufficient to induce a change in muscle phenotype. PMID- 15522233 TI - Identification of potential nuclear reprogramming and differentiation factors by a novel selection method for cloning chromatin-binding proteins. AB - Nuclear reprogramming is critical for animal cloning and stem cell creation through nuclear transfer, which requires extensive remodeling of chromosomal architecture involving dramatic changes in chromatin-binding proteins. To understand the mechanism of nuclear reprogramming, it is critical to identify chromatin-binding factors specify the reprogramming process. In this report, we have developed a high-throughput selection method, based on T7 phage display and chromatin immunoprecipitation, to isolate chromatin-binding factors expressed in mouse embryonic stem cells using primary mouse embryonic fibroblast chromatin. Seven chromatin-binding proteins have been isolated by this method. We have also isolated several chromatin-binding proteins involved in hepatocyte differentiation. Our method provides a powerful tool to rapidly and selectively identify chromatin-binding proteins. The method can be used to study epigenetic modification of chromatin during nuclear reprogramming, cell differentiation, and transdifferentiation. PMID- 15522234 TI - Effect of mutations in HNF-1alpha and HNF-1beta on the transcriptional regulation of human sucrase-isomaltase in Caco-2 cells. AB - Mutations in transcription factors hepatocyte nuclear factors (HNF)-1alpha and HNF-1beta cause maturity-onset diabetes of the young (MODY) types 3 and 5, respectively. HNF-1alpha and HNF-1beta mutations are well studied in some tissues, but the mechanism by which HNF-1alpha and HNF-1beta mutations affect sucrase-isomaltase (SI) transcription in the small intestine is unclear. We studied the effects of 13 HNF-1alpha mutants and 2 HNF-1beta mutants on human SI gene transcription, which were identified in subjects with MODY3 and MODY5, respectively. Transactivation activity of 11 HNF-1alpha and 2 HNF-1beta mutants was significantly lower than that of wild (wt)-HNF-1alpha and wt-HNF-1beta. Furthermore, in co-expression studies with mutant (mu)-HNF-1alpha/ wt-HNF-1beta and wt-HNF-1alpha/mu-HNF-1beta, the combination of mu-HNF-1alpha (P379fsdelCT and T539fsdelC)/wt-HNF-1beta impaired SI transcription, but the others were not remarkably different from wt-HNF-1alpha/wt-HNF-1beta. Although wt-HNF-1beta inhibited the transactivation activity of wt-HNF-1alpha on SI transcription, the inhibitory effect was reduced by 2 HNF-1beta mutants. These results suggest that SI transcription might tend to be unchanged or lower in MODY3, while occurring more in MODY5. PMID- 15522235 TI - Tyrosine phosphatase epsilonM stimulates migration and survival of porcine aortic endothelial cells by activating c-Src. AB - The cell growth, survival, and migration of vascular endothelial cells (ECs) are positively regulated by several protein tyrosine kinase receptors. Therefore, protein tyrosine phosphatases (PTPs) must also be important for these processes. The present study found that transmembranal PTPepsilonM, but not cytoplasmic PTPepsilonC, is expressed in porcine ECs and in rat smooth muscle cells, both of which were prepared from the aorta. The overexpression of wild-type PTPepsilonM promoted cell survival and migration in porcine aortic ECs even in medium without and with 1% serum, respectively. A catalytically inactive, substrate-trapping mutant of PTPepsilonM, respectively, did not affect and conversely suppressed cell survival and migration. Interestingly, the forced expression of wild-type PTPepsilonC reduced cell viability in contrast to PTPepsilonM in ECs lacking endogenous PTPepsilonC, indicating the biological significance of selective expression of PTPepsilon isoforms in the vasculature. PTPepsilonM activated c-Src kinase probably by directly dephosphorylating phospho-Tyr527, a negative regulatory site of c-Src. The increases in cell survival and migration induced by overexpressed PTPepsilonM were suppressed by the c-Src inhibitor SU6656. Considering the behaviors of vascular ECs in the pathogenesis of atherosclerosis, these data suggest that PTPepsilonM negatively regulates the development of this disease by activating c-Src. PMID- 15522236 TI - Glutamate receptor-mediated regulation of c-fos expression in cultured microglia. AB - It has been recently shown that the expression of various types of neurotransmitter receptors is not restricted to neurons but also observed in a majority of glial cells. However, their function in glial cells is not known well in both physiological and pathological conditions. Here, we investigated the role of glutamate receptor on c-fos gene expression in primary cultured and BV-2 microglia. Our results demonstrated that both c-fos mRNA and protein were dramatically induced following treatment with various glutamate receptor agonists (500muM); N-methyl-d-aspartic acid, kainic acid, (S)-alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid, and (RS)-3,5-dihydroxyphenylglycine. The responses were significantly suppressed by specific antagonists and also by calcium chelating agents EGTA and BAPTA-AM. Our results suggest that glutamate receptor activation regulates c-fos gene expression by modifying intracellular calcium levels in microglia. These findings might provide an insight in to understanding the function of microglial glutamate receptors in neuron-to-glial interaction under the excitotoxic conditions. PMID- 15522237 TI - Type I collagen synergistically enhances PDGF-induced smooth muscle cell proliferation through pp60src-dependent crosstalk between the alpha2beta1 integrin and PDGFbeta receptor. AB - Smooth muscle cells (SMCs) are exposed to both platelet-derived growth factor (PDGF) and type I collagen (CNI) at the time of arterial injury. In these studies we explore the individual and combined effects of these agonists on human saphenous vein SMC proliferation. PDGF-BB produced a 5.5-fold increase in SMC DNA synthesis whereas CNI stimulated DNA synthesis to a much lesser extent (1.6-fold increase). Alternatively, we observed an 8.3-fold increase in DNA synthesis when SMCs were co-incubated with CNI and PDGF-BB. Furthermore, stimulation of SMCs with PDGF-BB produced a significant increase in ERK-2 activity whereas CNI alone had no effect. Co-incubation of SMCs with PDGF-BB and CNI resulted in ERK-2 activity that was markedly greater than that produced by PDGF-BB alone. In a similar fashion, PDGF-BB induced phosphorylation of the PDGF receptor beta (PDGFRbeta) and CNI did not, whereas concurrent agonist stimulation produced a synergistic increase in receptor activity. Blocking antibodies to the alpha2 and beta1 subunits eliminated this synergistic interaction, implicating the alpha2beta1 integrin as the mediator of this effect. Immunoprecipitation of the alpha2beta1 integrin in unstimulated SMCs followed by immunoblotting for the PDGFRbeta as well as Src family members, pp60(src), Fyn, Lyn, and Yes demonstrated coassociation of alpha2beta1 and the PDGFRbeta as well as pp60(src). Incubation of cells with CNI and/or PDGF-BB did not change the degree of association. Finally, inhibition of Src activity with SU6656 eliminated the synergistic effect of CNI on PDGF-induced PDGFRbeta phosphorylation suggesting an important role for pp60(src) in the observed receptor crosstalk. Together, these data demonstrate that CNI synergistically enhances PDGF-induced SMC proliferation through Src-dependent crosstalk between the alpha2beta1 integrin and the PDGFRbeta. PMID- 15522238 TI - Sphingosine-1-phosphate lyase SPL is an endoplasmic reticulum-resident, integral membrane protein with the pyridoxal 5'-phosphate binding domain exposed to the cytosol. AB - Sphingosine-1-phosphate (S1P) is a sphingolipid metabolite that functions as a bioactive lipid molecule. S1P is degraded either by S1P lyase or by S1P phosphohydrolase. The gene encoding mammalian S1P lyase, SPL, has been identified. Here, we characterize the SPL protein in its expression, localization, and topology. The expression levels of the SPL protein correlated well with the dihydrosphingosine-1-phosphate (DHS1P) lyase activity in most tissues. However, liver and heart exhibited high DHS1P lyase activities compared to their SPL protein levels. The SPL mRNA expression was temporally regulated during mouse embryonal development. Immunofluorescence microscopy demonstrated that SPL is localized at the endoplasmic reticulum. Proteinase K digestion studies revealed that the large hydrophilic domain, containing the active site, faces the cytosol. This active site orientation is opposite to that of S1P phosphohydrolase, indicating that the degradation of S1P by two S1P-degrading enzymes occurs in spatially separated sides of the endoplasmic reticulum. PMID- 15522239 TI - alpha-Hemolysin-induced dephosphorylation of EGF receptor of A431 cells is carried out by rPTPsigma. AB - Earlier we have shown that the epidermal growth factor receptor was unable to retain its phospho Tyr signal after the assembly of staphylococcal alpha hemolysin (alpha-HL). However, the nature of the protein tyrosine phosphatase (PTPase) or its identity is not known. In this report, we demonstrate that the alpha-HL elevates the activity of receptor like protein tyrosine phosphatase sigma (rPTPsigma). The alpha-HL induced dephosphorylation is prominent only in intact A431 cells. The PTPase activity is not inhibited if the alpha-HL treatment precedes PTPase inhibitor treatments. The anti-EGFr immunoprecipitates have exhibited higher PTPase activity after alpha-HL treatment of A431 cells. Interestingly, PTPase activity of anti-EGFr immunoprecipitates from the A431 cells expressing the antisense message of rPTPsigma has not increased despite alpha-HL treatment, confirming the role of rPTPsigma in the dephosphorylation of EGFr. The studies presented here will be useful in understanding the process of signal modulation by the assembly of alpha-HL. PMID- 15522240 TI - Novel motifs in amino acid permease genes from Leishmania. AB - Eight amino acid permease genes from the protozoan parasite Leishmania donovani (AAPLDs) were cloned, sequenced, and shown to be expressed in promastigotes. Seven of these belong to the amino acid transporter-1 and one to the amino acid polyamino-choline superfamilies. Using these sequences as well as known and characterized amino acid permease genes from all kingdoms, a training set was established and used to search for motifs, using the MEME motif discovery tool. This study revealed two motifs that are specific to the genus Leishmania, four to the family trypanosomatidae, and a single motif that is common between trypanosomatidae and mammalian systems A1 and N. Interestingly, most of these motifs are clustered in two regions of 50-60 amino acids. Blast search analyses indicated a close relationship between the L. donovani and Trypanosoma brucei amino acid permeases. The results of this work describe the cloning of the first amino acid permease genes in parasitic protozoa and contribute to the understanding of amino acid permease evolution in these organisms. Furthermore, the identification of genus-specific motifs in these proteins might be useful to better understand parasite physiology within its hosts. PMID- 15522241 TI - A single amino acid substitution differentiates Hsp70-dependent effects on alpha synuclein degradation and toxicity. AB - alpha-Synuclein aggregation and toxicity play a major role in Parkinson's disease and dementia with Lewy bodies. Hsp70 is a multipurpose stress response chaperone protein that mediates both refolding and degradation of misfolded proteins. We have shown that Hsp70 is able to block both alpha-synuclein toxicity and aggregation. Here we introduce a mutation into the ATPase domain of Hsp70 (K71S) and demonstrate that this abolishes Hsp70 refolding activity. Nonetheless, Hsp70K71S continues to mediate alpha-synuclein degradation and blocks aggregate formation. In contrast to wild type Hsp70, the ATPase domain mutant mediates alpha-synuclein degradation through a non-proteasome inhibitor sensitive pathway. Although Hsp70K71S can diminish levels of alpha-synuclein to an even greater extent than Hsp70, HSP70K71S does not protect against alpha-synuclein toxicity. The Hsp70K71S mutant appears to dissociate the formation of aggregates, which it blocks, and toxicity, which it does not block. These data suggest that the ability of Hsp70 to prevent toxicity is distinct from degradation of alpha synuclein and is dependent on its ATPase domain. PMID- 15522242 TI - Expression of SARS-coronavirus envelope protein in Escherichia coli cells alters membrane permeability. AB - To promote viral entry, replication, release, and spread to neighboring cells, many cytolytic animal viruses encode proteins responsible for modification of host cell membrane permeability and for formation of ion channels in host cell membranes during their life cycles. In this study, we show that the envelope (E) protein of severe acute respiratory syndrome-associated coronavirus can induce membrane permeability changes when expressed in Escherichia coli. E protein expressed in bacterial and mammalian cells under reducing conditions existed as monomers, but formed homodimer and homotrimer under non-reducing conditions. Site directed mutagenesis studies revealed that two cysteine residues of the E protein were essential for oligomerization, leading to induction of membrane permeability. This is the first report demonstrating that a coronavirus-encoded protein could modify membrane permeability in E. coli cells. PMID- 15522243 TI - New approaches for exploring anatomical and functional connectivity in the human brain. AB - Information processing in the primate brain is based on the complementary principles of modular and distributed information processing. The former emphasizes the specialization of functions within different brain areas. The latter emphasizes the massively parallel nature of brain networks and the fact that function also emerges from the flow of information between brain areas. The localization of function to specific brain areas ("functional segregation") is the commonest approach to investigating function; however, an emerging, complementary approach ("functional integration") describes function in terms of the information flow across networks of areas. Here, we highlight recent advances in neuroimaging methodology that have made it possible to investigate the anatomical architecture of networks in the living human brain with diffusion tensor imaging (DTI). We also highlight recent thinking on the ways in which functional imaging can be used to characterize information transmission across networks in the human brain (functional and effective connectivity). PMID- 15522244 TI - Structural neural networks subserving oculomotor function in first-episode schizophrenia. AB - BACKGROUND: Smooth pursuit and antisaccade abnormalities are well documented in schizophrenia, but their neuropathological correlates remain unclear. METHODS: In this study, we used statistical parametric mapping to investigate the relationship between oculomotor abnormalities and brain structure in a sample of first-episode schizophrenia patients (n = 27). In addition to conventional volumetric magnetic resonance imaging, we also used magnetization transfer ratio, a technique that allows more precise tissue characterization. RESULTS: We found that smooth pursuit abnormalities were associated with reduced magnetization transfer ratio in several regions, predominantly in the right prefrontal cortex. Antisaccade errors correlated with gray matter volume in the right medial superior frontal cortex as measured by conventional magnetic resonance imaging but not with magnetization transfer ratio. CONCLUSIONS: These preliminary results demonstrate that specific structural abnormalities are associated with abnormal eye movements in schizophrenia. PMID- 15522245 TI - Repetitive transcranial magnetic stimulation reveals abnormal plastic response to premotor cortex stimulation in schizophrenia. AB - BACKGROUND: Schizophrenia may be characterized by abnormal plastic modulation in cortical neuronal circuits. Activation of premotor cortex using repetitive transcranial magnetic stimulation (rTMS) produces suppression of cortical excitability in primary motor cortex. We hypothesized that premotor rTMS would cause less suppression of motor cortical excitability in patients with schizophrenia than in control subjects. METHODS: Twelve patients diagnosed with schizophrenia and twelve healthy control subjects underwent subthreshold rTMS to the premotor area in a 15-min conditioning train. Measurements of primary motor cortical excitability (motor evoked potential; MEP), the resting motor threshold (RMT), and cortical inhibition (CI) were taken before and after the rTMS. RESULTS: There was no difference in RMT between groups at baseline, although the patient group had less CI than the control group at baseline. Following rTMS, the change in both MEP size and RMT between groups was significant. After rTMS, MEP size was suppressed in the control group and increased in the patient group, whereas RMT increased in the normal control group and decreased in the patient group. CONCLUSIONS: Patients with schizophrenia demonstrate abnormal brain responses to rTMS applied to the premotor cortex that appear to relate to reduced motor cortical inhibition. PMID- 15522246 TI - Preconditioning with transcranial direct current stimulation sensitizes the motor cortex to rapid-rate transcranial magnetic stimulation and controls the direction of after-effects. AB - BACKGROUND: Rapid-rate repetitive transcranial magnetic stimulation (rTMS) can produce a lasting increase in cortical excitability in healthy subjects or induce beneficial effects in patients with neuropsychiatric disorders; however, the conditioning effects of rTMS are often subtle and variable, limiting therapeutic applications. Here we show that magnitude and direction of after-effects induced by rapid-rate rTMS depend on the state of cortical excitability before stimulation and can be tuned by preconditioning with transcranial direct current stimulation (tDCS). METHODS: Ten healthy volunteers received a 20-sec train of 5 Hz rTMS given at an intensity of individual active motor threshold to the left primary motor hand area. This interventional protocol was preconditioned by 10 min of anodal, cathodal, or sham tDCS. We used single-pulse TMS to assess corticospinal excitability at rest before, between, and after the two interventions. RESULTS: The 5-Hz rTMS given after sham tDCS failed to produce any after-effect, whereas 5-Hz rTMS led to a marked shift in corticospinal excitability when given after effective tDCS. The direction of rTMS-induced plasticity critically depended on the polarity of tDCS conditioning. CONCLUSIONS: Preconditioning with tDCS enhances cortical plasticity induced by rapid-rate rTMS and can shape the direction of rTMS-induced after-effects. PMID- 15522248 TI - Circadian regulation of cortisol after hippocampal damage in humans. AB - BACKGROUND: There is substantial evidence that the hippocampus (HC) regulates the activity of the hypothalamic-pituitary-adrenocortical (HPA) axis. Damage to the HC in animals produces a transient alteration in diurnal and stress-related HPA activity. This study was designed to examine the effects of HC damage on basal cortisol secretion in humans. METHODS: Salivary cortisol was measured in 22 patients with HC damage (12 with bilateral damage and 10 with unilateral damage), 7 brain-damaged comparison participants, 10 healthy, age-matched comparison participants, and 6 of the patients' caregivers. Salivary cortisol samples were taken immediately after awakening, 30 min after awakening, at 8:00 am, 11:00 am, 3:00 pm, 6:00 pm, and at bedtime on a single day. Brain-injured patients underwent a structural magnetic resonance imaging scan to examine quantitative volumes of the HC. RESULTS: Both bilateral and unilateral HC damage abolished the cortisol response to awakening documented in the comparison groups. Caregivers of bilateral HC patients showed a reduced response to awakening. The remainder of the circadian pattern was not affected in the HC patients; all groups showed a significant diurnal variation. There was no association between HC volume and cortisol secretion. CONCLUSIONS: Hippocampal damage in humans abolishes the cortisol response to awakening, whereas the remainder of the diurnal cycle is unaffected in these patients. These data suggest a unique role of the HC in the control of basal cortisol secretion. PMID- 15522247 TI - Cellular changes in the postmortem hippocampus in major depression. AB - BACKGROUND: Imaging studies report that hippocampal volume is decreased in major depressive disorder (MDD). A cellular basis for reduced hippocampal volume in MDD has not been identified. METHODS: Sections of right hippocampus were collected in 19 subjects with MDD and 21 normal control subjects. The density of pyramidal neurons, dentate granule cell neurons, glia, and the size of the neuronal somal area were measured in systematic, randomly placed three-dimensional optical disector counting boxes. RESULTS: In MDD, cryostat-cut hippocampal sections shrink in depth a significant 18% greater amount than in control subjects. The density of granule cells and glia in the dentate gyrus and pyramidal neurons and glia in all cornv ammonis (CA)/hippocampal subfields is significantly increased by 30%-35% in MDD. The average soma size of pyramidal neurons is significantly decreased in MDD. CONCLUSION: In MDD, the packing density of glia, pyramidal neurons, and granule cell neurons is significantly increased in all hippocampal subfields and the dentate gyrus, and pyramidal neuron soma size is significantly decreased as well. It is suggested that a significant reduction in neuropil in MDD may account for decreased hippocampal volume detected by neuroimaging. In addition, differential shrinkage of frozen sections of the hippocampus suggests differential water content in hippocampus in MDD. PMID- 15522249 TI - High-affinity nicotinic acetylcholine receptors are required for antidepressant effects of amitriptyline on behavior and hippocampal cell proliferation. AB - BACKGROUND: A wide variety of antidepressants act as noncompetitive antagonists of nicotinic acetylcholine receptors (nAChRs), but the relationship between this antagonism and the therapeutic effects of antidepressants is unknown. METHODS: Antidepressant properties of the noncompetitive nAChR antagonist mecamylamine in the forced swim test were tested alone and in combination with the tricyclic antidepressant amitriptyline. Mice lacking high-affinity nAChRs were tested in three behavioral models to determine whether these receptors are required for behavioral effects of amitriptyline in common models of antidepressant action. Finally, the brains of wild-type and knockout animals treated with amitriptyline were examined to determine whether high-affinity nAChRs are required for antidepressant-induced increases in hippocampal cell proliferation. RESULTS: Inhibition of nAChRs by mecamylamine had antidepressant-like effects in the forced swim test and potentiated the antidepressant activity of amitriptyline when the two drugs were used in combination. Mice lacking high-affinity nAChRs showed no behavioral response to amitriptyline. Finally, after chronic treatment with amitriptyline, nAChR knockout mice did not show the increase in hippocampal cell proliferation seen in wild-type mice. CONCLUSIONS: These data support the hypothesis that antagonism of nAChRs is an essential component of the therapeutic action of antidepressants. PMID- 15522250 TI - The dopamine-4 receptor gene associated with binge eating and weight gain in women with seasonal affective disorder: an evolutionary perspective. AB - BACKGROUND: We recently described a preliminary association between the hypofunctional seven-repeat allele of the dopamine-4 receptor gene (DRD4) and increased maximal lifetime body mass index in women with seasonal affective disorder (SAD). In this study, we examined whether binge eating behavior mediated this putative association. METHODS: The study sample consisted of 131 women with winter SAD who reported increased intake of high-carbohydrate/high-fat foods during depressive episodes. We compared rates of binge eating behavior in the two genotypic groups defined by the presence or absence of the seven-repeat allele of DRD4. RESULTS: Consistent with our working hypothesis, the proportion of binge eaters was significantly greater in probands with the seven-repeat allele (18 of 46, 39.1%) than in probands without this allele (14 of 85, 16.5%) [chi(2)(1)= 8.32, p = .004; odds ratio = 3.25, 95% confidence interval 1.43, 7.41]. CONCLUSIONS: Pending replication in other samples, these results point to a genetic factor that could help in the early identification and treatment of women at higher risk for seasonal weight gain associated with binge eating behavior. At a theoretic level, the current results suggest a novel link between evolutionary models of seasonal weight gain on the one hand and the DRD4 gene on the other. PMID- 15522251 TI - Cerebrospinal fluid beta-amyloid1-42 and tau in control subjects at risk for Alzheimer's disease: the effect of APOE epsilon4 allele. AB - BACKGROUND: Cerebrospinal fluid (CSF) measures of beta-amyloid(1-42) and tau are linked with the known neuropathology of Alzheimer's disease (AD). Numerous lines of evidence have also suggested that individuals with at least one APOE epsilon4 allele on chromosome 19 are at increased risk of developing AD. We tested these CSF markers in groups of subjects with AD and healthy older control subjects, using the absence or presence of the APOE epsilon4 allele as a predictive variable in the search for possible prognostic biomarkers of AD. METHODS: We assessed the levels of beta-amyloid(1-42) and total tau in the CSF of 292 subjects (142 control subjects and 150 subjects with mild-to-moderate AD), who were research participants at the National Institute of Mental Health. The group of control subjects was enriched with a high percentage of subjects with a positive family history of AD. All subjects underwent extensive global cognitive testing. RESULTS: When divided according to the absence or presence of the APOE epsilon4 allele, the control subjects with at least one epsilon4 allele had significantly lower CSF beta-amyloid(1-42) but not tau levels than control subjects without an APOE epsilon4 allele (p < .01). As expected, the AD patients had lower levels of CSF beta-amyloid(1-42) and higher CSF tau levels than the normal control group (p < .01). CONCLUSIONS: The association of APOE epsilon4 allele and lower, more AD-like levels of CSF beta-amyloid(1-42) in older control subjects is consistent with previous studies showing possible neuroimaging and cognitive abnormalities with epsilon4 carriers and suggests that CSF beta amyloid(1-42) decreases might represent an early biomarker of AD. Longitudinal follow-up is of course required to verify whether this biomarker is indeed predictive of clinical conversion to AD. PMID- 15522252 TI - Catechol-O-methyltransferase val108/158met genotype predicts working memory response to antipsychotic medications. AB - BACKGROUND: The gene encoding catechol-O-methyltransferase (COMT), an enzyme that regulates prefrontal cortex dopamine, contains a common functional polymorphism (val(108/158)met) that influences prefrontal cortex function in an allelic dose dependent manner. A recent study reported that the COMT val(108/158)met polymorphism influences cognitive- and physiologic-related prefrontal cortex responses to antipsychotic treatment. The present study tested the effects of several COMT polymorphisms on the cognitive response to antipsychotic medication in patients with schizophrenia. METHODS: Twenty inpatients with schizophrenia or schizoaffective disorder (5 with the val-val genotype, 11 with val-met, and 4 with met-met) were administered cognitive tests at two time points: once after 4 weeks of treatment with antipsychotic medication and once after 4 weeks of placebo administration, according to a counterbalanced, double-blind, within subject study design. RESULTS: Patients homozygous for the COMT met allele displayed significant improvement on the working memory task after treatment. Patients homozygous for the COMT val allele did not show working memory improvement with treatment. Other COMT polymorphisms were not associated with significant differences between treatment and placebo conditions. CONCLUSIONS: These results support other data suggesting that the COMT val(108/158)met polymorphism might be an important factor in the cognitive response to antipsychotic medication. PMID- 15522253 TI - Association analysis of FEZ1 variants with schizophrenia in Japanese cohorts. AB - BACKGROUND: DISC1 has been suggested as a causative gene for psychoses in a large Scottish family. We recently identified FEZ1 as an interacting partner for DISC1. To investigate the role of FEZ1 in schizophrenia and bipolar disorder, case control association analyses were conducted in Japanese cohorts. METHODS: We performed a mutation screen of the FEZ1 gene and detected 15 polymorphisms. Additional data on informative polymorphisms were obtained from public databases. Eight single nucleotide polymorphisms (SNPs) were analyzed in 119 bipolar disorder and 360 schizophrenic patients and age- and gender-matched control subjects. All genotypes were determined with the TaqMan assay, and selected samples were confirmed by sequencing. RESULTS: The two adjacent polymorphisms displayed a nominally significant association with schizophrenia (IVS2+ 1587G>A, p = .014; 396T50% less than adhesion to laminin. Inhibition of HAS2 and/or HAS3 or pretreatment with hyaluronidase significantly decreased adhesion of SW620 cells to laminin (P < 0.05), suggesting that adhesion was dependent upon pericellular HA. CONCLUSIONS: Metastatic SW620 cells that produce large amounts of pericellular HA adhered well to laminin. Inhibition of HAS2 and/or HAS3 expression, or hyaluronidase digestion of pericellular HA significantly inhibited adhesion. These data suggest that HA promotes adhesion to laminin and may thereby facilitate invasion of the basement membrane and metastasis in colon carcinoma. PMID- 15522318 TI - Hepatocytes derived from peripheral blood stem cells of granulocyte-colony stimulating factor treated F344 rats in analbuminemic rat livers. AB - BACKGROUND: hematopoietic stem cells (SCs) mobilized from the bone marrow (BM) into peripheral blood (PB) are reported to have ability to differentiate into various cell types. We investigated whether PB-SCs mobilized by treatment with granulocyte-colony stimulating factor (G-CSF) in normal rats can raise albumin producing hepatocytes after transplantation within the liver of analbuminemic rats. MATERIALS AND METHODS: Fischer 344 rats (F344) were used as donors, and F344 congenic Nagase's analbuminemic rats (F344alb) as recipients. The donors were repeatedly treated with human recombinant G-CSF, and their PB mononuclear cells (MNCs) were infused into the portal veins of recipients immediately after 70% hepatectomy (PH). RESULTS: Although a few single and small clusters (less than five cells) of albumin positive (alb+) hepatocytes were seen in the livers of untreated F344alb and of the animals undergoing PH alone or transplantation of PB-MNCs with or without the prior G-CSF treatment, clusters consisting of more than 6 alb+ hepatocytes were only detected in the livers of recipients that received transplantation of mobilized PB-MNCs or BM-MNCs under the regenerating condition induced by PH. Sry3, a Y chromosome marker, could be detected corresponding to the alb+ clusters by in situ hybridization when male donors and female recipients were used. Moreover, normal albumin gene sequences were demonstrated in the microdissected alb+ clusters by polymerase chain reaction, and the serum albumin levels were elevated in the recipients. CONCLUSIONS: Hematopoietic SCs mobilized from BM into PB by the G-CSF treatment may raise hepatocyte colonies, when transplanted into regenerating livers. PMID- 15522319 TI - Perioperative diagnosis of cystosarcoma phyllodes of the breast may be enhanced by MIB-1 index. AB - BACKGROUND: The recurring theme in cystosarcoma phyllodes (CSP) is one of underdiagnosis by pathologists and undertreatment by surgeons. Major areas of investigation relating to the diagnosis of CSP center on accurate preoperative diagnosis, elucidating the relevance of histological classification with respect to outcome, and identifying novel markers to reliably differentiate CSP from fibroadenoma (FA). MATERIALS AND METHODS: Fifteen CSP and 7 cellular FA controls (where the preoperative diagnosis was unclear) were retrospectively investigated. Preoperative histological and radiological investigations were reviewed for efficacy. The ability of MIB-1 antibody to differentiate the two fibroepithelial lesions was investigated using immunohistochemical estimation of the MIB-1 index. RESULTS AND DISCUSSION: Preoperative core biopsy had a sensitivity of 75% but was carried out in only 23% of cases. Fine needle aspiration cytology and radiological assessment were not efficacious in preoperative diagnosis. Proliferative activity (MIB-1 indices) was significantly higher in CSP than in a selected population of FA where there was preoperative diagnostic uncertainty (P < 0.0001). Indices were also able to determine CSP subclassification. This suggests MIB-1 as a constructive adjunctive investigation when evaluating histological features to differentiate CSP from FA in difficult cases. CONCLUSIONS: The use of MIB-1 may increase the sensitivity of preoperative core biopsy diagnosis, offering more effective surgical planning and decreasing immediate reoperation rates. PMID- 15522320 TI - Usefulness of inhibiting the lymph node metastasis in human gastric carcinoma by B7-1 gene transfection. AB - INTRODUCTION: Lymph node metastasis is one of the crucial prognostic factors in gastric cancer. We have reported that ICAM-1 gene transfection was effective against lymph node metastases of gastric cancer. B7-1, one of the co-stimulatory factors, was reported to induce cytotoxic T lymphocytes when using melanoma and bladder cancer cell lines, as well as ICAM-1. In this study, we investigated the inhibitory effect of B7-1 on lymph node metastasis by B7-1 gene transfection into gastric cancer cells. MATERIALS AND METHODS: We transfected B7-1 genes into a gastric cancer cell line (OCUM-2MLN) and analyzed the effect of B7-1 transduction on lymph node metastasis, the in vitro adhesiveness and cytotoxicity assay of mononuclear lymphocytes to cancer cells and lymph node metastatic ability after orthotopic implantation of gastric cancer cells in vivo. RESULTS: We revealed that mononuclear lymphocytes showed significantly stronger adherence and cytotoxicity to B7-1 transfected cells (2MLN/B7) than its parent OCUM-2MLN cells. The tumor growth rate of 2MLN/B7 xenograft was significantly slower than OCUM 2MLN xenograft in nude mice. In orthotopic implantation experiments for nude mice, 2MLN/B7 cells in stomach developed significantly less lymph node metastasis than OCUM-2MLN cells. Histologic findings showed that leukocytes were intensively infiltrated in both the 2MLN/B7 tumors and its metastatic lesions, however, were scarcely observed in the lesions associated with 2MLN cells. CONCLUSION: B7-1 may play an important role in inhibiting lymph node metastasis by the mechanism of enhanced immunogenicity, and that B7-1 gene transduction might be effective against lymph node metastases of gastric cancer. PMID- 15522321 TI - Ergothioneine pretreatment protects the liver from ischemia-reperfusion injury caused by increasing hepatic heat shock protein 70. AB - BACKGROUND: Reperfusion of the liver after ischemia induces the expression of the heat shock genes and the synthesis of the heat shock proteins (HSP). We studied the effects of the natural antioxidant ergothioneine (EGT) treatment on the expression of HSP70 in ischemic-reperfused (IR) liver. METHODS: Adult male Wistar rats were randomly divided into three groups: Sham group given standard laboratory chow and water for 3 weeks followed by sham operation; Control group given standard laboratory chow and water for 3 weeks followed by liver IR injury; EGT group given standard laboratory chow supplementation l-ergothioneine (1.2 mg/kg/d body weight) administered by gavage and water for 3 weeks followed by liver IR injury. Ten rats from each group were killed to determine serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactic dehydrogenase (LDH), tissue malondialdehyde (MDA), HSP70 levels, and histologic changes at 30, 60, and 120 min of reperfusion, respectively. Survival was followed for 1 week. RESULTS: IR caused significant increase in serum AST, ALT, LDH, and tissue MDA levels. As compared with the control group, animals treated with EGT experienced a significant decrease in serum AST, ALT, and LDH levels in all reperfusion periods. Tissue MDA levels in animals receiving EGT were significantly reduced as compared with control group at 30 min and 60 min after reperfusion. After ischemia, reperfusion caused a remarkable production of HSP70 in the control group. When the rats were pretreated with EGT, the levels of HSP70 increased significantly in their livers after reperfusion compared with the control group. Liver injury in the EGT-treated animals was lower to that in the control group. The 7-day survival rate was significantly improved (from 50% to 80%) by EGT pretreatment. CONCLUSION: HSP70 has been shown to induce tolerance against warm IR injury in rat livers. EGT pretreatment protects the liver from IR injury by over-expression of HSP and the subsequent suppression of lipid peroxidation. PMID- 15522322 TI - Depletion of activated Vbeta8+ T cells disrupts bispecific antibody directed antitumor immunity. AB - INTRODUCTION: Activation of Vbeta8+ T cells with superantigen staphylococcal enterotoxin B (SEB) and use of an antitumor, anti-CD3 bispecific antibody (BsAb) leads to tumor protective immunity. We hypothesize that Vbeta8+ T-cell activation in combination with BsAb is crucial for tumor protective immunity in this model. METHODS: Adolescent C3H/HeN mice were intravenously injected with syngeneic CL62 melanoma to establish pulmonary metastasis. Three days after establishing pulmonary metastasis, predominantly Vbeta8+ T cells are activated with 50 mug of intraperitoneal superantigen SEB. T cells were depleted at different time points in relation to SEB administration to assess the effect on protective immunity against a second tumor challenge. RESULTS: Protective immunity is significantly (P < 0.008) decreased when Vbeta8+ depletion occurs 6 h after SEB injection, as growth of rechallenged CL62 melanoma occurred in 43%. Protective immunity is present at all other time points when mice survive Vbeta8+ T-cell depletion. Survival of animals treated with SEB/BsAb (82%) is significantly better (P < 0.002) than with SEB alone (60%) or nontreated control (0%). Survival when Vbeta8+ T-cell depletion occurred at 6 h and 48 h post-SEB is 72% and 77%, respectfully, and is statistically indistinguishable (P < 0.232 and P < 0.602). If T-cell depletion was conducted before SEB administration, however, the combination of SEB and BsAb did not result in significant protective immunity. T cell depletion before the use of SEB alone, without BsAb, failed to result in significant protective immunity. CONCLUSIONS: Depletion of Vbeta8+ T cells 6 h after activation disrupts the development of protective immunity. PMID- 15522323 TI - Dose dependent effects of cardiac beta2 adrenoceptor gene therapy. AB - BACKGROUND: Adenoviral-mediated gene transfer during cardiopulmonary bypass (CPB) achieves efficient myocardial transgene expression. The optimal vector dose required to produce not only increased beta adrenoceptor (betaAR) density but, more importantly, enhanced left ventricular (LV) function is unknown. In addition, it is unclear if absent extracardiac expression in preliminary studies represented cardiac specific, as opposed to selective gene delivery, as a consequence of low vector doses. MATERIALS AND METHODS: Adenoviral vector encoding the human beta(2) adrenoceptor (Adeno-beta(2)AR) was delivered to cardioplegic arrested hearts of neonatal piglets during CPB in three doses ranging from 5 x 10(11) total viral particles (tvp) to 2 x 10(12) tvp. Control animals received adenoviral vector encoding beta galactosidase (Adeno-betagal) or PBS (PBS). LV and liver betaAR density and in vivo LV function were assessed 5 days later. RESULTS: Elevated LV betaAR density was present after delivery of Adeno-beta(2)AR at all doses. Piglets which received 5 x 10(11) tvp and 1 x 10(12) tvp Adeno-beta(2)AR demonstrated enhanced LV dP/dt(max) but in those receiving 2 x 10(12) tvp LV dP/dt(max) was unchanged. Moreover, at this higher dose of adenoviral vector the detrimental effects of cardiac inflammation and extracardiac gene overexpression became apparent. CONCLUSIONS: Although the highest increase in cardiac betaAR density occurred after high-dose Adeno beta(2)AR, LV dP/dt(max) was not enhanced. Moreover, significant extracardiac gene expression was present at this dose, emphasizing the need for careful dose response studies in gene therapy. However, cardiac selective beta(2)AR overexpression does occur following adenoviral vector delivery during CPB and cardioplegic arrest resulting in enhanced LV dP/dt(max). PMID- 15522324 TI - Thrombus-induced endothelial dysfunction: Hemoglobin and fibrin decrease nitric oxide bioactivity without altering eNOS. AB - BACKGROUND: Arterial thrombosis is associated with endothelial dysfunction. The purpose of this study was to determine which components of thrombus induce endothelial dysfunction and to determine their effect on endothelial nitric oxide synthase (eNOS) activity and expression. MATERIALS AND METHODS: Human aortic endothelial cells were grown to confluence. Group 1 consisted of control cells exposed to media. Group 2 was exposed to cellular components of thrombus, including erythrocytes (RBCs) (1.5, 3, and 6 x 10(4) cells/ml) and platelets (0.5, 1, and 2 x 10(5) platelets/ml). Group 3 was exposed to extracellular thrombus components, including hemoglobin (1.25, 2.5, and 5.0 g/dl), thrombin (0.4, 4.0, and 10.0 units/ml), and fibrin. The exposure time was 20 h. Nitric oxide (NO) levels were measured following exposure. Global cellular integrity was evaluated by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4 sulfophenyl)-2H-tetrazolium) conversion. eNOS mRNA expression was measured using semiquantitative PCR and eNOS protein activity evaluated via a l-citrulline conversion assay. Studies were done in replicates of six and expressed as percentages of controls. RESULTS: NO levels decreased following exposure to hemoglobin (1.25 g/dl = 59 +/- 5%, 2.5 g/dl = 38 +/- 3%, 5 g/dl = 37 +/- 6%, P < 0.001). MTS metabolism was likewise suppressed (1.25 g/dl = 57 +/- 2%, 2.5 g/dl = 55 +/- 3%, 5 g/dl = 44 +/- 6%, P < 0.001). Fibrin diminished NO levels (37 +/- 5%, P < 0.01) and MTS metabolism (49 +/- 5%, P < 0.01). RBCs and platelets had no effect on NO production or MTS metabolism (P > 0.05). Thrombin's effect was concentration dependent, inhibiting nitric oxide production at low doses while stimulating it at high doses (P < 0.01). All thrombin doses enhanced MTS metabolism (P < 0.05). eNOS activity was not altered by fibrin, thrombin, or hemoglobin exposure (P > 0.05). Moreover, eNOS mRNA expression was unaffected (P > 0.05). CONCLUSIONS: Intact cellular components of thrombus do not cause endothelial dysfunction in vitro. However, free hemoglobin and fibrin diminish NO bioactivity, likely via scavenging. Thrombin affects NO levels in a concentration dependent manner. Neither eNOS expression nor eNOS activity is diminished, indicating that thrombus does not cause permanent changes in NO generation capability. PMID- 15522325 TI - Effect of an EGF-cancer vaccine on wound healing and inflammation models. AB - BACKGROUND: Epidermal growth factor (EGF) and its receptor (EGF-R) are attractive targets for cancer immunotherapy. Tolerance has been broken with an EGF-vaccine and antibodies against EGF have been produced in animals and in cancer patients. EGF also plays an important role in the inflammation stage of wound healing. Because this therapeutic approach may be of importance after surgery procedures in cancer patients, we decided to investigate the possible role of the EGF vaccine in the croton-oil-induced ear edema and in the wound healing experimental animal models. MATERIALS AND METHODS: Mice were immunized with an EGF-vaccine by intramuscular injections and serum titers against EGF were measured through ELISA techniques. Control animals received saline. RESULTS: Immunized mice produced antibodies against EGF while no antibody titers could be measured in control animals. Croton oil applied to the inner ear surface of EGF-vaccine treated mice caused a 61.3% lower ear punch weight and a 60.2% lower myeloperoxidase activity than control mice. In the EGF-vaccine treated animals, planimetry measurements and histological analysis did not led to significant impairment in tissue repair. CONCLUSIONS: The EGF-vaccination in mice decreased the normal croton-oil-induced inflammation response, without apparent impairment in tissue healing. PMID- 15522326 TI - The role of thrombospondin-1 in human disease. AB - Thrombospondin-1 (TSP-1) is a large matricellular glycoprotein secreted by many cell types. It is a component of the extracellular matrix during active and subacute processes. Due to TSP-1's ability to interact with a variety of matrix proteins and cell-surface receptors, controversy exists about its conflicting functions. In this review, we will discuss the role of TSP-1 in human disease. PMID- 15522327 TI - Benzo[a]pyrene removal from soil by Phanerochaete chrysosporium grown on sugarcane bagasse and pine sawdust. AB - The capacity of Phanerochaete chrysosporium grown on soil with added sugarcane baggase (BP) and pine sawdust (PS) to remove benzo(a)pyrene (BaP) was studied. A half factorial two-level experiment 2(4-1) was designed to determine the effect of: type of lignocellulosic material (BP and PS) for fungus growth, age of fungus (5 and 10d), amount of lignocellulosic material (10% and 15% w/w) and soil moisture content (water holding capacity of 45% and 56% w/w). Inoculum obtained at different ages showed that the capacity of P. chrysosporium to remove BaP depends on the lignocellulosic used and on inoculum age. Abiotic BaP removal was affected significantly (p<0.05) by inoculum age, type of lignocellulosic added and soil moisture content. The removal of BaP by lignocellulosic material was more effective by young inocula (71.97 mg BaP kg(-1) dry soil), with high percentage of added lignocellulosic (71.57 mg BaP kg(-1) dry soil) and at low soil moisture content (73.07 mg BaP kg(-1) dry soil). When fungus was grown on BP, maximum BaP removal rate was obtained at 5d of incubation (10.85 mg BaP d( 1)l(-1) and 50.12 mg BaP kg(-1) dry soil), while in PS maximum BaP removal was obtained at 10d of incubation (12.06 mg BaP d(-1)l(-1) and 39.94 mg BaP kg(-1) dry soil). PMID- 15522328 TI - Oxidation of methane over palladium catalysts: effect of the support. AB - This work is focused on the deep catalytic oxidation of methane over supported palladium catalysts. The influences of the metal loading, oxidation state of palladium, nature of supports, presence of promoters in the supports (for zirconia-based supports), and thermal stability have been studied experimentally. Catalysts were prepared by incipient wetness of commercially available supports with aqueous solutions of palladium nitrate. For gamma-alumina support, it was observed that the optimal amount of palladium is between 0.5% and 2%, with higher amounts leading to a loss in specific activity. Concerning the oxidation state of the catalyst, it is concluded that for all the supports tested in the present work, a reduction of the catalyst is not needed, yielding the same conversion at steady state catalysts reduced and oxidised. The thermal stability of various supported catalysts were also studied, zirconia supports being the most active. These supports, specially Y-modified zirconia support, do not suffer appreciable deactivation below 500 degrees C. PMID- 15522329 TI - Polyacrylamide sorption opportunity on interlayer and external pore surfaces of contaminant barrier clays. AB - Physico-chemical interactions among polymer molecules in aqueous solution and clay mineralogical/textural characteristics influence the sorption of polymer molecules on clay barrier minerals. Amendment of potentially unstable barrier clays with aqueous polymers can improve barrier material resistance to environmental stresses during service. In this research, the ability of molecular coils of polyacrylamide (PAM) to overlap in solution and to enter interlayer space in Na-montmorillonite (specific surface=31.82+/-0.22 m2 g(-1)) and kaolinite (specific surface=18+/-2 m2 g(-1)) were analyzed theoretically and experimentally, using solution viscosity measurements, and X-ray diffractometry. Experimental data on two theoretical indices: relative size ratio (RSR); and molecular availability (Ma) that are formulated to scale polymer molecular sorption on clay interlayer, indicate that the sorption of PAM A (Mw=4000000) and PAM B (Mw=7000000) does not produce any significant change in the d-spacing of both clay minerals. Although the negative Ma values of -3.51 g l(-1) for PAM A and -3.88 g l(-1) for PAM B indicate high levels of entanglement of polymer molecular coils in solution, sorption data confirm that the entangled coils are still able to sorb onto Na-montmorillonite highly and kaolinite to a lesser extent. PMID- 15522330 TI - Solubilization and biodegradation of phenanthrene in mixed anionic-nonionic surfactant solutions. AB - The effects of mixed anionic-nonionic surfactants, sodium dodecyl sulfate (SDS) mixed with Tween80 (TW80), Triton X-100 (TX100) and Brij35 respectively on the solubility enhancement and biodegradation of phenanthrene in the aqueous phase were investigated. The efficiency of solubilization and biodegradation of phenanthrene in single-, and mixed-surfactant solutions were also compared. The critical micellar concentrations (CMCs) of mixed surfactants were sharply lower than that of sole SDS. The degree of solubility enhancements by the mixed surfactants followed the order of SDS-TW80>SDS-Brij35>SDS-TX100. Synergistic solubilization was observed in the mixed surfactant solutions, in which the molar ratios of SDS to nonionic surfactant were 1:0, 9:1, 7:3, 5:5, 3:7, 1:9 and 0:1 while the total concentration of surfactants was kept at 5.0 and 10.0 mM, respectively. SDS-Brij35 exhibited more significant degree of synergistic solubility enhancement for phenanthrene. The mixed surfactants exhibited no inhibitory effect on biodegradation of phenanthrene. Substantial amounts of the solubilized phenanthrene by mixed surfactants were completely degraded by phenanthrene-degrading microorganisms within 96 h. The results suggested that anionic-nonionic surfactants would improve the performance of remediation of PAH contaminated soils. PMID- 15522331 TI - Electrodegradation of landfill leachate in a flow electrochemical reactor. AB - Sanitary landfills are the major method used today for the disposal and management of municipal solid waste. Decomposition of waste and rainfall generate leachate at the bottom of landfills, causing groundwater contamination. In this study, leachate from a municipal landfill site was treated by electrochemical oxidation in a pilot scale flow reactor, using oxide-coated titanium anode. The experiments were conducted under a constant flow rate of 2000 lh(-1) and the effect of current density on chemical oxygen demand, total organic carbon, color and ammonium removal was investigated. At a current density of 116.0 mA cm(-2) and 180 min of processing, the removal rates achieved were 73% for COD, 57% for TOC, 86% for color and 49% for ammonium. The process proved effective in degrading leachate, despite this effluent's usual refractoriness to treatment. PMID- 15522332 TI - Improvement of coagulation-flocculation process using anionic polyacrylamide as coagulant aid. AB - A physicochemical treatment (coagulation-flocculation) was applied to a slaughterhouse wastewater, using anionic polyacrylamide as coagulant aid to improve the settling velocity of the flocs formed with the coagulants used: ferric sulphate, aluminium sulphate and polyaluminium chloride. The optimum speed and stirring time for the flocculation stage were ascertained along with the optimum pH and coagulant and coagulant aid doses. The speed and coagulation time were initially set according to recommendations in the literature concerning the treatment of this type of water. Chemical oxygen demand (COD), biochemical oxygen demand at 5 days (BOD5) and total suspended solids (TSS) were recorded at the beginning and end of each experiment in order to monitor the process. Once the optimal conditions had been established, several parameters were measured in order to assess the coagulation-flocculation process: particle number and size, sludge volume, nutrients (ammonia nitrogen, total Kjeldahl nitrogen, albuminoid nitrogen, orthophosphate, total phosphorus) and the residual concentration of iron and aluminium in clarified water. Anionic polyacrylamide, when added with ferric sulphate or polyaluminium chloride led to a significant increase in the settling speed. PMID- 15522333 TI - Biological solubilization and mineralization as novel approach for the pretreatment of food waste. AB - The performance of a garbage disposal system to solubilize and mineralize food wastes through biological solubilization was evaluated through the examination of the effects of operational conditions like water supply volume, water supply frequency and aeration on the amount of waste solubilized, mineralized and accumulated in the reactor. The biological solubilization process consisted of a solubilization reactor and a circulation tank. Food waste and fresh water were supplied into the solubilization reactor with support media. Wastewater from the solubilization reactor was discharged to the circulation tank and water in the circulation tank was periodically pumped back to the solubilization reactor. In case of the total food waste loading of 16 kg m(3-1) d(-1), little carbon (0 5.7%) accumulated in the reactor as long as the system was kept under aerobic condition through large volume of water supply (higher than 3.5 lh(-1)) or applying aeration in the circulation tank. However, 42% of the loaded carbon accumulated under anaerobic condition in low water supply (less than 1.8 lh(-1)). The rest of the waste was either solubilized or mineralized. The aeration in the circulation tank, therefore, was effective to provide similar solubilization and mineralization as the large volume of water supply. However, frequency of feeding at the large volume of water supply had no significant effect on the amount of waste solubilization and mineralization. PMID- 15522334 TI - Chromium(VI) sorptive removal from aqueous solutions by nanocrystalline akaganeite. AB - In this study, akaganeite (beta-FeO(OH)) an ironoxyhydroxide material, was used as a low-cost potential adsorbent for the removal of hexavalent chromium from aqueous solutions. The influence of agitation speed, solution pH, initial chromium concentration, sorbent concentration and temperature were evaluated at batch kinetic runs. It was shown that the solid diffusion model, in comparison to simple reaction kinetic models, described better the sorption kinetics. Freundlich and Frumkin isotherm best fitted the equilibrium results. Akaganeite presented a sorption capacity approximately 80 mg Cr(VI) g(-1), under the conditions studied. Flotation was used as a downstream process for the effective removal of the loaded material. PMID- 15522335 TI - Assessment of the mechanical stability and chemical leachability of immobilized electroplating waste. AB - The effectiveness of cement based treatment technology, in immobilizing chromium laden electroplating sludge was assessed by conducting toxicity characteristic leaching procedure (TCLP). The mechanical stability of the blocks was tested by measuring the compressive strength. Other leaching tests such as NEN 7341 test, ANS 16.1 and multiple TCLP (MTCLP) test conducted on select solidified blocks showed that chromium was immobilized by the binder studied. A linear relationship was obtained between the cumulative fraction of chromium leached (CFL) and square root of time in the solidified samples proving that diffusion is the controlling mechanism for leaching of chromium. The leachability indices (LI) obtained for the solidified materials using cement and cement-fly ash system (EPC6, EPFC6A and EPFC6B) satisfy the guidance value as per US NRC, which clearly indicates that chromium is well retained within the solid matrix. Chromium concentrations in the TCLP leachates of the above mix ratios were well within the regulatory level of United States Environmental Protection Agency (USEPA). Molecular characterization of the solidified material was carried out using Fourier transformation infra red (FTIR) technique. PMID- 15522336 TI - Bioremediation potential of a perchlorate-enriched sewage sludge consortium. AB - The purpose of this work was to explore the reductive bioremediation potential of a perchlorate-enriched facultative anaerobic consortium. Rapid perchlorate reduction and bacterial growth were observed up to 1.84 g l(-1) of perchlorate, but not at 3.82 g l(-1) due to the toxicity. The specific growth rate of the mixed consortium was 0.1 h(-1). The consortium co-reduced perchlorate and nitrate with acetate as e- donor and carbon source. The presence of nitrate slowed down the perchlorate reduction rate. The other e- acceptors utilized include oxygen, chlorate, Cr(VI), and selenate. Over 95% of the 16 mg l(-1) of added Cr(VI) was reduced within 24 h of incubation with a high-density perchlorate-grown consortium. However, the consortium failed to couple growth with reduction of nitrite, sulfate, thiosulfate, and sulfite. During the search for autotrophic perchlorate reduction, many consortia from very diverse natural sources could not use sulfur compounds such as thiosulfate as e- donor. PMID- 15522337 TI - Selenium extractability from a contaminated fine soil fraction: implication on soil cleanup. AB - Two batches of fine soil fraction of an acidic soil were deliberately contaminated with selenite (Se(IV)) and selenate (Se(VI)), respectively, and aged for more than 220 days. Speciation analysis using continuous flow-through hydride generation atomic absorption spectrometry (HGAAS) indicated that the species were predominant in their respective aged soils. A selective sequential extraction scheme was employed to fractionate the Se retained in the soils into six fractions of varying retentions. Abilities of various chemical reagents in extracting the Se in the two soil batches were then evaluated. The reagents investigated were sodium salts such as sodium chloride (NaCl), sodium sulfate (Na2SO4), sodium carbonate (Na2CO3), and sodium phosphate (Na3PO4), and two oxidants, namely, hydrogen peroxide (H2O2) and potassium permanganate (KMnO4). It was found that NaCl, Na2SO4, and Na2CO3 could only extract the exchangeable fraction of Se, while Na3PO4 could extract the exchangeable and strongly-bound fractions. Selenate was extracted more than Se(IV) by the salts. The kinetics of Se(IV) extraction by Na3PO4 could be best described by the Elovich model, while the Ritchie second-order model was the most appropriate to describe Se(VI) extraction. Efficiencies of the oxidants in Se(IV) extraction highly depended on their applied dosages. Both H2O2 and KMnO4 were able to extract greater than 93% of total Se, and therefore were significantly more effective than the salts in Se(IV) extraction. PMID- 15522338 TI - Iron and organo-bentonite for the reduction and sorption of trichloroethylene. AB - Hybrid barriers using dechlorination and immobilization were studied to remove trichloroethylene (TCE) in this study. Hybrid barriers of iron filings and organo (hexadecyltrimethylammonium, HDTMA)-bentonite were simulated in columns to assess the performance of the hybrid barriers. TCE reduction rate for the mixture of zero valent iron (ZVI) and HDTMA-bentonite was approximately seven times higher than that for ZVI, suggesting the reduction of TCE was accelerated when HDTMA bentonite was mixed with ZVI. For the column of two separate layers of iron and HDTMA-bentonite, TCE reduction rate was nearly similar to that for ZVI alone, but the partition coefficient (Kd) was 4.5 times higher than that for ZVI only. TCE was immobilized in the first layer with HDTMA-bentonite due to sorption, and then dechlorinated in the second layer with iron filings due to reduction. The HDTMA bentonite and minimally-desorbed HDTMA from the organo-bentonite are believed to contribute the increase in TCE concentration on iron surface so that more TCE could be available for reduction. Therefore, the incorporation of HDTMA-bentonite into ZVI not only can effectively retard the transport of chlorinated organic contaminants from landfill leachate or oil shock in subsurface environment, also can expedite the reduction rate of TCE. PMID- 15522339 TI - Adsorption of nitrosamines in acidic solution by zeolites. AB - Adsorption of N-nitrosopyrrolidine (NPYR) by zeolite NaZSM-5, Hbeta and NaZSM-11 in acidic solution with pH of 1 was investigated. The NPYR uptake process obeyed the first-rate Lagegren kinetic equation describing adsorption, and the equilibrium data were fitted to the Langmuir or Freundlich adsorption equation. Zeolite ZSM-5, Hbeta and NaZSM-11 possessed a higher adsorption capacity than amorphous silica or ordered mesoporous siliceous SBA-15, implying their potential application for adsorption of nitrosamines in gastric juice to anti-cancer. PMID- 15522340 TI - Modification of Langmuir isotherm in solution systems--definition and utilization of concentration dependent factor. AB - The Langmuir isotherm, originally derived for the adsorption of gas molecules on solid surfaces, was modified to fit the adsorption isotherm of solutes onto solid surfaces in solution systems. The aim of this modification is based on the fact that direct application of the Langmuir isotherm to solution systems often leads to poor data fitting. In the present communication, it is shown that the level of data fitting to the Langmuir isotherm of literature data can be improved by a simple modification introducing a concentration dependent factor, X. The key concept of the modification lies in that the concentration of solute affects both adsorption and desorption stages. As a first approximation, we adopted a single term polynomial for both processes of adsorption and desorption. Based on reanalysis of literature data of adsorption in solution, we confirmed that indeed the modified Langmuir isotherm more accurately describes the experimental observations. Furthermore, we proposed that the concentration dependent factor could be associated with the surface heterogeneity index that was introduced in a few other modified Langmuir isotherms. Some advantages and limitations of proposed modified Langmuir isotherm are also discussed. PMID- 15522341 TI - Lead biosorption and desorption by intact and pretreated spirulina maxima biomass. AB - In order to search for locally available and untried biomaterials in China with high removal capacity of heavy metals from wastewater, the feasibility of Spirulina maxima as biosorbent for lead removal and recovery from aqueous solution was investigated. The lead biosorption was studied by using intact biomass and pretreated biomass of S. maxima. The effects of operational conditions (e.g. pH, contact time, biomass concentration etc.) on lead biosorption were investigated. The biosorption was solution pH dependent and the maximum adsorption was obtained at a solution pH of about 5.5. The adsorption equilibrium was reached in 60 min. The biosorption followed the Freundlich isotherm model. The maximum removal ratios of lead were about 84% in intact biomass and 92% in pretreated biomass. The lead adsorbed could be desorbed effectively by 0.1 M nitric acid, EDTA and hydrochloric acid. The results in this study indicated that pretreated biomass of S. maxima was a promising candidate for removing lead from wastewater. PMID- 15522342 TI - Feedback-mediated dynamics in two coupled nephrons. AB - Previously, we developed a dynamic model for the tubuloglomerular feedback (TGF) system in a single, short-looped nephron of the mammalian kidney. In that model, a semi-linear hyperbolic partial differential equation was used to represent two fundamental processes of solute transport in the nephron's thick ascending limb (TAL): chloride advection by fluid flow along the TAL lumen and transepithelial chloride transport from the lumen to the interstitium. An empirical function and a time delay were used to relate glomerular filtration rate to the chloride concentration at the macula densa of the TAL. Analysis of the model equations indicated that stable limit-cycle oscillations (LCO) in nephron fluid flow and chloride concentration can emerge for sufficiently large feedback gain magnitude and time delay. In this study, the single-nephron model was extended to two nephrons, which were coupled through their filtration rates. Explicit analytical conditions were obtained for bifurcation loci corresponding to two special cases: (1) identical time delays but differing feedback gains, and (2) identical gains but differing delays. Similar to the case of a single nephron, our analysis indicates that stable LCO can emerge in coupled nephrons for sufficiently large gains and delays. However, these LCO may emerge at lower values of the feedback gain, relative to a single (i.e., uncoupled) nephron, or at shorter delays, provided the delays are sufficiently close. These results suggest that, in vivo, if two nephrons are sufficiently similar, then coupling will tend to increase the likelihood of LCO. PMID- 15522343 TI - Homeostasis of peripheral immune effectors. AB - In this paper, we use both mathematical modeling and simulation to explore homeostasis of peripheral immune system effector cells, particularly alveolar macrophages. Our interest is in the distributed control mechanisms that allow such a population to maintain itself. We introduce a multi-purpose simulator designed to study individual cell responses to local molecular signals and their effects on population dynamics. We use the simulator to develop a model of growth factor regulation of macrophage proliferation and survival. We examine the effects of this form of regulation in the context of two competing hypotheses regarding the source of new alveolar macrophages. In one model, local cells divide to replenish the population; in the other, only cells migrating from circulation divide. We find that either scenario is plausible, although the influx-driven system is inherently more stable. The proliferation-driven system requires lower cell death and efflux rates than the influx-driven system. PMID- 15522344 TI - Characterizing fluorescence recovery curves for nuclear proteins undergoing binding events. AB - Fluorescence recovery after photobleaching (FRAP) is an experimental technique used to measure the mobility of proteins within the cell nucleus. After proteins of interest are fluorescently tagged for their visualization and monitoring, a small region of the nucleus is photobleached. The experimental FRAP data are obtained by recording the recovery of the fluorescence in this region over time. In this paper, we characterize the fluorescence recovery curves for diffusing nuclear proteins undergoing binding events with an approximate spatially homogeneous structure. We analyze two mathematical models for interpreting the experimental FRAP data, namely a reaction-diffusion model and a compartmental model. Perturbation analysis leads to a clear explanation of two important limiting dynamical types of behavior exhibited by experimental recovery curves, namely, (1) a reduced diffusive recovery, and (2) a biphasic recovery characterized by a fast phase and a slow phase. We show how the two models, describing the same type of dynamics using different approaches, relate and share common ground. The results can be used to interpret experimental FRAP data in terms of protein dynamics and to simplify the task of parameter estimation. Application of the results is demonstrated for nuclear actin and type H1 histone. PMID- 15522345 TI - Evolutionary tradeoff and equilibrium in an aquatic predator-prey system. AB - Due to the conventional distinction between ecological (rapid) and evolutionary (slow) timescales, ecological and population models have typically ignored the effects of evolution. Yet the potential for rapid evolutionary change has been recently established and may be critical to understanding how populations persist in changing environments. In this paper we examine the relationship between ecological and evolutionary dynamics, focusing on a well-studied experimental aquatic predator-prey system (Fussmann et al., 2000, Science, 290, 1358-1360; Shertzer et al., 2002, J. Anim. Ecol., 71, 802-815; Yoshida et al., 2003, Nature, 424, 303-306). Major properties of predator-prey cycles in this system are determined by ongoing evolutionary dynamics in the prey population. Under some conditions, however, the populations tend to apparently stable steady-state densities. These are the subject of the present paper. We examine a previously developed model for the system, to determine how evolution shapes properties of the equilibria, in particular the number and identity of coexisting prey genotypes. We then apply these results to explore how evolutionary dynamics can shape the responses of the system to 'management': externally imposed alterations in conditions. Specifically, we compare the behavior of the system including evolutionary dynamics, with predictions that would be made if the potential for rapid evolutionary change is neglected. Finally, we posit some simple experiments to verify our prediction that evolution can have significant qualitative effects on observed population-level responses to changing conditions. PMID- 15522346 TI - Sliding window discretization: a new method for multiple band matching of bacterial genotyping fingerprints. AB - Microbiologists have traditionally applied hierarchical clustering algorithms as their mathematical tool of choice to unravel the taxonomic relationships between micro-organisms. However, the interpretation of such hierarchical classifications suffers from being subjective, in that a variety of ad hoc choices must be made during their construction. On the other hand, the application of more profound and objective mathematical methods--such as the minimization of stochastic complexity--for the classification of bacterial genotyping fingerprints data is hampered by the prerequisite that such methods only act upon vectorized data. In this paper we introduce a new method, coined sliding window discretization, for the transformation of genotypic fingerprint patterns into binary vector format. In the context of an extensive amplified fragment length polymorphism (AFLP) data set of 507 strains from the Vibrionaceae family that has previously been analysed, we demonstrate by comparison with a number of other discretization methods that this new discretization method results in minimal loss of the original information content captured in the banding patterns. Finally, we investigate the implications of the different discretization methods on the classification of bacterial genotyping fingerprints by minimization of stochastic complexity, as it is implemented in the BinClass software package for probabilistic clustering of binary vectors. The new taxonomic insights learned from the resulting classification of the AFLP patterns will prove the value of combining sliding window discretization with minimization of stochastic complexity, as an alternative classification algorithm for bacterial genotyping fingerprints. PMID- 15522347 TI - Fuzzy modeling in symptomatic HIV virus infected population. AB - This paper introduces a model for the evolution of positive HIV population and manifestation of AIDS (acquired immunideficiency syndrome). The focus is on the nature of the transference rate of HIV to AIDS. Expert knowledge indicates that the transference rate is uncertain and depends strongly on the viral load and the CD4+ level of the infected individuals. Here, we suggest to view the transference rate as a fuzzy set of the viral load and CD4+ level values. In this case the dynamic model results in a fuzzy model that preserves the biological meaning and nature of the transference rate lambda. Its behavior fits the natural history of HIV infection reported in the medical science domain. The paper also includes a comparison between the fuzzy model and a classic Anderson's model using data reported in the literature. PMID- 15522348 TI - Evolutionary game dynamics in finite populations. AB - We introduce a model of stochastic evolutionary game dynamics in finite populations which is similar to the familiar replicator dynamics for infinite populations. Our focus is on the conditions for selection favoring the invasion and/or fixation of new phenotypes. For infinite populations, there are three generic selection scenarios describing evolutionary game dynamics among two strategies. For finite populations, there are eight selection scenarios. For a fixed payoff matrix a number of these scenarios can occur for different population sizes. We discuss several examples with unexpected behavior. PMID- 15522349 TI - Resistance is useless?--Extensions to the game theory of kleptoparasitism. AB - We extend the game theoretic model of kleptoparasitism introduced by Broom and Ruxton (1998, Behav. Ecol. 9, 397-403) in two ways: we allow for asymmetric contests, where the probability alpha of the challenger winning can take any value from 0 to 1; and we allow the handler to choose not to resist the challenge, but to immediately concede and relinquish its food to the challenger. We find, in general, three possible evolutionarily stable strategies--challenge and-resist (Hawk), challenge-but-do-not-resist (Marauder) and do-not-challenge but-resist (Retaliator). When alpha = 1/2, we find that Hawk and Marauder are the only ESS's, in contrast to the result of the original model; we also find an overlap region, in parameter space, where two different ESS's are possible, depending on initial conditions. For general alpha, we see that all three ESS are possible, depending on different values of the environmental parameters; however, as the average time of a contest over food becomes long, then the Marauder strategy becomes more and more prevalent. The model makes a potentially significant prediction about animal behaviour in the area of kleptoparasitism, that a searcher, when it meets a handler, will only decline to attack that handler when alpha < 1/2 i.e. when the defender is more likely to win. One possible converse of this statement, that a handler whose probability of success is greater than 1/2 should always resist a challenge, is not true. PMID- 15522350 TI - Seasonal population dynamics of ticks, and its influence on infection transmission: a semi-discrete approach. AB - In this paper, a simple semi-discrete (ticks' feeding is assumed to occur only during the summers of each year) model for tick population dynamics is presented. Conditions for existence, uniqueness, and stability of a positive equilibrium are found; the system is then studied numerically using parameter estimates calibrated for the tick Ixodes ricinus in Trentino, Italy, and the sensitivity to parameters is examined. Then, this model is extended to consider a tick transmitted infection of one species of hosts, while other hosts are incompetent to the infection. Assuming, for simplicity, that the infection is not affecting the total number either of hosts or ticks, a threshold condition for infection persistence is obtained. The dependence of the equilibrium infection prevalence on parameters is studied numerically; in particular, we considered how infection prevalence depends on host densities. This analysis reveals that a 'dilution effect' occurs both for competent and for incompetent hosts; this means that, besides a lower threshold for host densities for infection to persist, there exists also an upper threshold: if host densities were higher than the upper threshold, the infection would go to extinction. Numerically, it is found that, for realistic parameter values, the upper threshold is not much higher than observed densities. PMID- 15522351 TI - An analysis of a nonlinear stage-structured cannibalism model with application to the Northeast Arctic cod stock. AB - A two-dimensional stage-structured population model with nonlinear cannibalism terms is studied. We show that there is a large parameter interval where the nontrivial equilibrium of the model is the only stable attractor, but that there also exist parameter intervals where we find quasiperiodic, periodic and chaotic dynamics. Moreover, in the interplay between increasing the fecundity and increasing the cannibalism pressure, the former turns out to be a destabilizing effect while the latter tends to act in a stabilizing fashion. Finally, we have applied the model to the North Atlantic cod stock using ICES biomass estimates. Our main conclusion from this study is that the combined effect of recruitment and cannibalism may not serve as an explanation of the observed fluctuations in the cod stock. PMID- 15522352 TI - Mathematical aspects of protein structure determination with NMR orientational restraints. AB - The field of structural biology is becoming increasingly important as new technological developments facilitate the collection of data on the atomic structures of proteins and nucleic acids. The solid-state NMR method is a relatively new biophysical technique that holds particular promise for determining the structures of peptides and proteins that are located within the cell membrane. This method provides information on the orientation of the peptide planes relative to an external magnetic field. In this article, we discuss some of the mathematical methods and tools that are useful in deriving the atomic structure from these orientational data. We first discuss how the data are viewed as tensors, and how these tensors can be used to construct an initial atomic model, assuming ideal stereochemistry. We then discuss methods for refining the models using global optimization, with stereochemistry constraints treated as penalty functions. These two processes, initial model building followed by refinement, are the two crucial steps between data collection and the final atomic model. PMID- 15522353 TI - Resource allocation, hyperphagia and compensatory growth. AB - Organisms often shown enhanced growth during recovery from starvation, and can even overtake continuously fed conspecifics (overcompensation). In an earlier paper (Ecology 84, 2777-2787), we studied the relative role played by hyperphagia and resource allocation in producing overcompensation in juvenile (non reproductive) animals. We found that, although hyperphagia always produces growth compensation, overcompensation additionally requires protein allocation control which routes assimilate preferentially to structure during recovery. In this paper we extend our model to cover reproductively active individuals and demonstrate that growth rate overcompensation requires a similar combination of hyperphagia and allocation control which routes the part of enhanced assimilation not used for reproduction preferentially towards structural growth. We compare the properties of our dynamic energy budget model with an earlier proposal, due to Kooijman, which we extend to include hyperphagia. This formulation assumes that the rate of allocation to reserves is controlled by instantaneous feeding rate, and one would thus expect that an extension to include hyperphagia would not predict growth overcompensation. However, we show that a self-consistent representation of the hyperphagic response in Kooijman's model overrides its fundamental dynamics, leading to preferential allocation to structural growth during recovery and hence to growth overcompensation. PMID- 15522354 TI - Constructing rooted supertrees using distances. AB - Suppose that a family of rooted phylogenetic trees Ti with different sets Xi of leaves is given. A supertree for the family is a single rooted tree T whose leaf set is the union of all the Xi, such that the branching information in T corresponds to the branching information in all the trees Ti. This paper proposes a polynomial-time method BUILD-WITH-DISTANCES that makes essential use of distance information provided by the trees Ti to construct a rooted tree S0. When a supertree also containing the distance information exists, then S0 is a supertree. The supertree S0 often shows increased resolution over the trees found by methods that utilize only the topology of the input trees. When no supertree exists because the input trees are incompatible, several variants of the method are described which still produce trees with provable properties. PMID- 15522355 TI - Lattice and non-lattice models of tumour angiogenesis. AB - In order to progress from the relatively harmless avascular state to the potentially lethal vascular state, solid tumours must induce the growth of new blood vessels from existing ones, a process called angiogenesis. The capillary growth centres around endothelial cells: there are several cell-based models of this process in the literature and these have reproduced some of the key microscopic features of capillary growth. The most common approach is to simulate the movement of leading endothelial cells on a regular lattice. Here, we apply a circular random walk model to the process of angiogenesis, and thus allow the cells to move independently of a lattice; the results display good agreement with empirical observations. We also run simulations of two lattice-based models in order to make a critical comparison of the different modelling approaches. Finally, non-lattice simulations are carried out in the context of a realistic model of tumour angiogenesis, and potential anti-angiogenic strategies are evaluated. PMID- 15522356 TI - Comparison of three models predicting developmental milestones given environmental and individual variation. AB - In all organisms, phenotypic variability is an evolutionary stipulation. Because the development of poikilothermic organisms depends directly on the temperature of their habitat, environmental variability is also an integral factor in models of their phenology. In this paper we present two existing phenology models, the distributed delay model and the Sharpe and DeMichele model, and develop an alternate approach, called the Extended von Foerster model, based on the age structured McKendrick-von Foerster partial differential model. We compare the models theoretically by examining the biological assumptions made in the basic derivation of each approach. In particular, we focus on each model's ability to incorporate variability among individuals as well as variability in the environment. When compared against constant temperature mountain pine beetle (Dendroctonus ponderosae Hopkins) laboratory developmental data, the Extended von Foerster model exhibits the highest correlation between theory and observation. PMID- 15522357 TI - Cell directional persistence [corrected] and chemotaxis in vascular morphogenesis. AB - In vertebrates, supply of oxygen and nutrients to tissues is carried out by the blood vascular system through capillary networks. Capillary patterns are closely mimicked by endothelial cells cultured on Matrigel, a preparation of basement membrane proteins. On the Matrigel surface, single randomly dispersed endothelial cells self-organize into vascular networks. The network is characterized by a typical length scale, which is independent of the initial mean density of deposed cells n over a wide range of values of n. We give here a detailed description of a mathematical model of the process which is able to reproduce several qualitative and quantitative features of in vitro vascularization experiments. Cell matter is basically modelled as an elastic fluid subjected to a specific force field depending on the concentration of a chemoattractant factor. Starting from sparse initial data, mimicking the initial conditions realized in laboratory experiments, numerical solutions reproduce characteristic network structures, similar to observed ones, whose average size is theoretically related to the finite range of chemoattractant diffusion. A possible area of application of the model is the design of properly vascularized artificial tissues. PMID- 15522358 TI - Competitive regulation of plant allometry and a generalized model for the plant self-thinning process. AB - Taking into account the individual growth form (allometry) in a plant population and the effects of intraspecific competition on allometry under the population self-thinning condition, and adopting Ogawa's allometric equation 1/y = 1/axb + 1/c as the expression of complex allometry, the generalized model describing the change mode of r (the self-thinning exponential in the self-thinning equation, log M = K + log N, where M is mean plant mass, K is constant, and N is population density) was constructed. Meanwhile, with reference to the changing process of population density to survival curve type B, the exponential, r, was calculated using the software MATHEMATICA 4.0. The results of the numerical simulation show that (1) the value of the self-thinning exponential, r, is mainly determined by allometric parameters; it is most sensitive to change of b of the three allometric parameters, and a and c take second place; (2) the exponential, r, changes continuously from about -3 to the asymptote -1; the slope of -3/2 is a transient value in the population self-thinning process; (3) it is not a 'law' that the slope of the self-thinning trajectory equals or approaches -3/2, and the long-running dispute in ecological research over whether or not the exponential, r, equals -3/2 is meaningless. So future studies on the plant self-thinning process should focus on investigating how plant neighbor competition affects the phenotypic plasticity of plant individuals, what the relationship between the allometry mode and the self-thinning trajectory of plant population is and, in the light of evolution, how plants have adapted to competition pressure by plastic individual growth. PMID- 15522359 TI - Stability of cardiac waves. AB - Cardiac waves can fail to propagate when the membrane potential of the cells in the wavefront rises too slowly. The sodium channel inactivation gates play an important role in this process of propagation block. Simple models including inactivation gates can have travelling waves of constant form with two possible velocities. A stability analysis demonstrates that the slower velocity is always unstable, and in limited parameter regimes the faster velocity can also be unstable. Waves with the lower velocity propagate a finite distance before they dissipate due to this instability and this distance is calculated. The distance can be large suggesting that they might be seen in certain pathological conditions. The analytical results are compared with numerical simulations of the simplified model and a detailed cardiac ionic model. PMID- 15522360 TI - Synchrony & chaos in patchy ecosystems. AB - The apparent synchronisation of spatially discrete populations is a well documented phenomenon. However, it is not clear what the governing mechanisms are for this synchrony, and whether they are robust over a range of environmental conditions and patch specific population dynamic behaviours. In this paper, we explore two (possibly interacting) modes of coupling, and investigate their theoretically discernible, and perhaps even experimentally measurable, signatures. To aid us in this investigation we employ a planktonic example system, with direct application to plankton patchiness. Furthermore, we address the role of chaos in complex spatio-temporal dynamics; we find that chaos associated with funnel attractors can play a distinguished role, over dynamics less sensitive to small variations, in being more susceptible to generalised synchronisation (such as phase synchronisation) in the presence of small local parameter variation. This is in contrast to the case for coupled systems with identical dynamics, and suggests that non-identically coupled systems are more vulnerable to global extinction events when exhibiting funnel-type chaotic dynamics. PMID- 15522361 TI - Foldable intraocular lens removal/exchange: can it be prevented? PMID- 15522362 TI - Surgery for subfoveal choroidal neovascularization in age-related macular degeneration: ophthalmic findings: SST report no. 11. AB - PURPOSE: To present visual acuity (VA) and related findings from patients enrolled in one of the Submacular Surgery Trials (SST) evaluating surgical removal versus observation of subfoveal choroidal neovascularization secondary to age-related macular degeneration (SST Group N Trial). DESIGN: Randomized clinical trial. PARTICIPANTS: Eligible patients had age-related macular degeneration with subfoveal choroidal neovascularization, some with a classic pattern on fluorescein angiography, and best-corrected VA (BCVA) of 20/100 to 20/800 in one eye (study eye) that had received no treatment in the macula. Any contiguous blood had to account for <50% of the total area occupied by the subfoveal lesion (maximum size, 9.0 disc areas [22.9 mm2]). METHODS: Randomization was stratified by VA and by clinical center. All patients were scheduled for study examinations at 3, 6, 12, and 24 months after enrollment for assessment of study outcomes. MAIN OUTCOME MEASURE: A successful outcome was defined a priori to be either improvement of BCVA or VA no more than 1 line (7 letters) worse than baseline at the 24-month examination. RESULTS: Of 454 patients enrolled, 228 study eyes were assigned to observation and 226 to surgery. The percentages of eyes that had successful outcomes were similar in the 2 arms: 44% assigned to observation and 41% assigned to surgery. Median VA losses from baseline to the 24-month examination were 2.1 lines (10.5 letters) in the observation arm and 2.0 lines (10 letters) in the surgery arm. Median VA declined from 20/100 at baseline to 20/400 at 24 months in both arms. No subgroup of patients was identified in which submacular surgery led to better VA outcomes. In the surgery arm, 55 (39%) of 142 initially phakic eyes had cataract surgery by the 24-month examination, compared with 6 (5%) of 133 eyes in the observation arm. Rhegmatogenous retinal detachment occurred in 12 surgery eyes (5%) and 1 observation eye. CONCLUSIONS: Submacular surgery, as performed in this clinical trial, did not improve or preserve VA for 24 months in more eyes than observation and is not recommended for patients with similar lesions. This article contains additional online-only material available at http://www.ophsource.com/periodicals/ophtha. PMID- 15522363 TI - Surgery for subfoveal choroidal neovascularization in age-related macular degeneration: quality-of-life findings: SST report no. 12. AB - PURPOSE: To describe health-related quality of life (HRQOL), overall and in patients with unilateral or bilateral choroidal neovascularization (CNV), in a clinical trial (Group N Trial) comparing observation and surgical removal of subfoveal CNV secondary to age-related macular degeneration (AMD). DESIGN: Randomized clinical trial. PARTICIPANTS: Eligible patients had untreated subfoveal CNV and AMD, best-corrected visual acuity (VA) of 20/100 to 20/800, classic CNV on fluorescein angiography, and a total subfoveal lesion size of < or =9.0 disc areas in the study eye. METHODS: Health-related quality of life data (the National Eye Institute Visual Function Questionnaire [NEI-VFQ], 36-item Short Form Health Survey [SF-36], and Hospital Anxiety and Depression Scale [HADS]) and clinical data were collected at baseline and at 6, 12, 24, 36, and 48 months. Patients were divided into unilateral and bilateral CNV subgroups based on fluorescein angiographic and clinical evidence. MAIN OUTCOME MEASURE: Two-year change in the NEI-VFQ. RESULTS: Of 454 patients enrolled, 228 were assigned to observation and 226 to surgery. At baseline, median overall NEI-VFQ scores were 67 in the observation group and 69 in the surgery group; by 2 years, the observation group had lost a median of 3 points (95% confidence interval [CI]: -6 to -2), and the surgery group gained a median of 1 point (CI: -1 to 3). The largest difference was observed for the mental health subscale, where the observation group lost a median of 5 points (CI: -5 to 0), and the surgery group gained a median of 5 points (CI: 0-10) by 2 years. Treatment differences in median 2-year changes in NEI-VFQ scores favored surgery by up to 10 points for unilateral cases and up to 8 points for bilateral cases. No treatment difference in 2-year change was observed for the SF-36 physical component summary; 2-year change in the mental component summary favored surgery by 2 points. Few patients (2%-4%) had HADS definite anxiety or depression at baseline or at 24 months. CONCLUSIONS: Although HRQOL outcomes were better in the submacular surgery arm than in the observation arm, surgery (per protocol) is not recommended because VA outcomes (reported elsewhere) were similar in the treatment arms. This article contains additional online-only material available at http://www.ophsource.com/periodicals/ophtha. PMID- 15522364 TI - Surgery for hemorrhagic choroidal neovascular lesions of age-related macular degeneration: ophthalmic findings: SST report no. 13. AB - PURPOSE: To present best-corrected visual acuity (BCVA) findings and other clinical outcomes from eyes of patients enrolled in one of the Submacular Surgery Trials (SST) evaluating surgical removal versus observation of predominantly hemorrhagic subfoveal choroidal neovascularization (CNV) associated with age related macular degeneration. DESIGN: Randomized clinical trial (SST Group B Trial). PARTICIPANTS: Eligible patients had subfoveal choroidal neovascular lesions greater than 3.5 disk areas (8.9 mm2) composed of at least 50% blood (either blood or CNV underlying the center of the foveal avascular zone) and BCVA of 20/100 to light perception in the study eye. INTERVENTION: Patients were assigned randomly at time of enrollment to observation or surgical removal of blood and any associated CNV. MAIN OUTCOME MEASURE: A successful outcome was defined a priori as either improvement in visual acuity (VA), no change in VA, or a decline in VA of no more than 1 line (7 letters) from baseline to the 24-month examination based on an intent-to-treat analysis. RESULTS: Of 336 patients enrolled, 168 were assigned to each treatment arm; treatment arms were balanced by baseline characteristics. Of 1501 expected examinations 3 months through 36 months after baseline, 1370 (91%) were performed. Loss of > or =2 lines (> or =8 letters) of VA occurred in 56% of surgery eyes, versus 59% of observation eyes examined at 24 months. Although severe loss of VA was not the primary outcome of interest, surgery more often prevented such loss: 36% in the observation arm versus 21% in the surgery arm at the 24-month examination (chi2 P = 0.004). Of initially phakic eyes, the cumulative percentage that had undergone cataract surgery by 24 months was 44% in the surgery arm, compared with 6% in the observation arm. Twenty-seven eyes (16%) in the surgical arm, compared with 3 eyes (2%) in the observation arm, had a rhegmatogenous retinal detachment (RD). CONCLUSIONS: Submacular surgery as performed in the SST Group B Trial did not increase the chance of stable or improved VA (the primary outcome of interest) and was associated with a high risk of rhegmatogenous RD, but did reduce the risk of severe VA loss in comparison with observation. This article contains additional online-only material available at http://www.ophsource.com/periodicals/ophtha. PMID- 15522366 TI - Post-traumatic endophthalmitis. AB - OBJECTIVE: To establish risk factors for the occurrence of post-traumatic endophthalmitis, to observe the efficacy of prophylaxis, and to describe the clinical features of post-traumatic endophthalmitis. DESIGN: Partially prospective consecutive case-control study. PARTICIPANTS: A total of 250 consecutive patients admitted to a single ophthalmic hospital with open globe injuries during a 3-year period were included. METHODS: Patients with post traumatic endophthalmitis were identified prospectively and added to an endophthalmitis database. All open globe injuries during the same time period were identified through a retrospective search of inpatient admissions, and their charts were reviewed. Information collected from all patient files included patient age; gender; injury setting (indoor/outdoor); wound contamination; nature of injury (site on eye, lens involvement, retained intraocular foreign body); mechanism of injury (penetration/perforation/rupture/ruptured surgical wound); prophylactic antibiotic administration, including route and timing; timing of primary repair; lensectomy at the time of primary repair; and depot corticosteroid at the time of primary repair. Any association between these parameters and the subsequent development of endophthalmitis was investigated. Any association between endophthalmitis and final visual acuity (VA) and also enucleation was evaluated. MAIN OUTCOME MEASURE: Development of endophthalmitis. RESULTS: The frequency of endophthalmitis after open globe injury was 6.8%. The following factors were associated with the subsequent development of endophthalmitis by univariate analysis: dirty wound (14.3% vs. 4.1%, P = 0.01), retained intraocular foreign body (13.0% vs. 4.4%, P = 0.02), lens capsule breach (12.8% vs. 3.2%, P = 0.01), delayed primary repair (> or =12 hours) (11.3% vs. 2.9%, P = 0.02), and rural address (10.1% vs. 4.3%, P = 0.07). Risk factors identified after multivariate analysis were dirty injury (odds ratio [OR], 5.3; 95% confidence interval [CI)], 1.5-18.7), breach of lens capsule (OR, 4.4; 95% CI, 1.2-15.6), and delay in primary repair (per hour: OR, 1.013; 95% CI, 1.002 1.024). None of the following factors was found to be associated with post traumatic endophthalmitis: patient age, gender, injury setting, site of injury on eye, mechanism of injury, antibiotic administration, lensectomy at the time of primary repair, and depot corticosteroid at the time of primary repair. Final VA tended to be worse in eyes with endophthalmitis (P = 0.08). Endophthalmitis did not significantly influence the frequency of enucleation/evisceration (5.9% vs. 4.3%, P = 0.55). CONCLUSIONS: Delay in primary repair, ruptured lens capsule, and dirty wound were each independently associated with the development of post traumatic endophthalmitis. Patients with > or =2 of these 3 risk factors had a particularly high frequency of infection. PMID- 15522365 TI - Surgery for hemorrhagic choroidal neovascular lesions of age-related macular degeneration: quality-of-life findings: SST report no. 14. AB - PURPOSE: To present and compare findings from health-related quality-of-life (HRQOL) interviews conducted with patients enrolled in the SST Group B Trial evaluating surgical removal of subfoveal choroidal neovascular lesions associated with age-related macular degeneration versus observation. DESIGN: Randomized clinical trial. PARTICIPANTS: Eligible patients had predominantly hemorrhagic subfoveal choroidal neovascular lesions (total lesion size of >3.5 disc areas, area of blood at least 50% of the lesion area, and at least 75% of blood posterior to the equator) and best-corrected visual acuity (VA) of 20/100 to <20/1600 but at least light perception in the study eye. Three hundred thirty-six patients enrolled after baseline quality-of-life interviews, 168 assigned to each of surgery or observation. METHODS: Clinical and HRQOL data were collected before randomization and at 6, 12, 24, 36, and 48 months after enrollment. Baseline clinical evidence was used to stratify patients as having unilateral or bilateral neovascularization at the time of randomization. The HRQOL interviews included the National Eye Institute Visual Function Questionnaire (NEI-VFQ), the 36-item Short Form Health Survey, and the Hospital Anxiety and Depression Scale. MAIN OUTCOMES MEASURE: Two-year change in NEI-VFQ. RESULTS: At 24 months after enrollment, overall NEI-VFQ scores had a median decrease of 1 point from baseline in the observation arm (95% confidence interval [CI]: -4 to 3 points) and no change in the surgery arm (95% CI: -3 to 3 points) (P = 0.70). Changes from baseline on NEI-VFQ subscales also were similar between treatment arms. Differences in scores by unilateral or bilateral involvement seen at baseline in each treatment arm persisted throughout follow-up for most outcomes. Planned analyses stratified by VA showed trends (P = 0.17) in favor of surgery at 24 months in the patients with baseline VA greater than 20/200 for the NEI-VFQ scale (3.5-point median increase from baseline in the surgery arm [95% CI: -4 to 7] vs. a 1-point median loss from baseline in the observation arm [95% CI: -6 to 4]). CONCLUSIONS: No difference was detected with respect to vision-targeted quality of-life outcomes for patients randomized to surgery or observation in the SST Group B Trial. This article contains additional online-only material available at www.ophsource.com/periodicals/ophtha. PMID- 15522367 TI - Endophthalmitis caused by Haemophilus influenzae. AB - OBJECTIVE: To investigate the clinical settings, management strategies, antibiotic sensitivities, and visual acuity outcomes for eyes with endophthalmitis caused by Haemophilus influenzae. DESIGN: Retrospective, noncomparative, consecutive case series. METHODS: The medical records were reviewed of all patients treated for culture-proven H. influenzae endophthalmitis at a single institution between January 1, 1980, and December 31, 2002. MAIN OUTCOME MEASURES: Visual acuity and antibiotic sensitivities. RESULTS: The study included 16 eyes of 16 patients with a median age of 68 years (range, 6 months-83 years) and a median follow-up of 26 months (range, 2 months-15 years). Clinical settings included post-trabeculectomy (n = 7), post-cataract surgery (n = 6), post-pars plana vitrectomy (n = 1), post-secondary intraocular lens insertion (n = 1), and post-suture removal from an extracapsular cataract wound (n = 1). Eleven (69%) cases were of delayed onset (>6 weeks from surgery/event), with a median interval between surgery/event and presentation with endophthalmitis of 18 months (range, 44 days-21 years); 5 (31%) cases were of acute onset (median, 6 days; range, 2-14 days). Presenting visual acuity was hand movements or better in 7 (44%) eyes. A vitreous tap and inject was performed initially in 9 (56%) eyes, and a vitrectomy was performed initially in the remaining 7 (44%) eyes. All eyes received intravitreal antibiotics on the day of presentation, and 11 (69%) received intravitreal dexamethasone. In vitro testing of the H. influenzae isolates revealed that 14 of 16 (88%) were sensitive to vancomycin, ampicillin, and trimethoprim/sulfamethoxazole; 15 of 16 (94%) were sensitive to aminoglycosides (1 isolate was resistant to gentamicin); and all were sensitive to cephalosporins, fluoroquinolones, and carbapenems. The organisms were sensitive to at least 1 of the initial antibiotics administered in all cases. Final visual acuity was 5/200 or better in 6 (38%) eyes, and 6 (38%) eyes had a final visual acuity of no light perception. CONCLUSIONS: Endophthalmitis caused by H. influenzae is generally associated with poor visual outcomes despite prompt treatment with intravitreal antibiotics to which the organisms were sensitive. PMID- 15522368 TI - Stage 0 macular holes: observations by optical coherence tomography. AB - OBJECTIVE: To introduce the concept of a stage 0 macular hole based on optical coherence tomographic observations of the vitreoretinal interface in fellow eyes of patients with unilateral idiopathic macular holes, and to evaluate the subsequent risk of progression to a full-thickness macular hole. DESIGN: Retrospective observational case series. PARTICIPANTS: Ninety-four patients with a unilateral stage 2, 3, or 4 full-thickness macular hole. METHODS: The medical records of patients with a unilateral macular hole diagnosed between 1994 and 2000 at the New England Eye Center were reviewed. MAIN OUTCOME MEASURE: Development of a full-thickness macular hole in the fellow eye on biomicroscopic fundoscopy or optical coherence tomography (OCT). RESULTS: In 27 (28.7%) of 94 clinically normal fellow eyes, OCT detected an abnormality of the vitreoretinal interface but normal foveal anatomy. The vitreoretinal abnormalities were further subclassified into severe (4 eyes), moderate (8 eyes), and mild (15 eyes) based on the intensity and morphology of the OCT signal. One of the 4 (25%) severe cases progressed to a full-thickness macular hole, 4 of the 8 (50%) moderate cases became full-thickness macular holes, and no (0%) mild cases progressed to a full-thickness macular hole. Severe and moderate eyes seemed to share characteristic features on OCT that increased their risk of macular hole development (stage 0 macular hole). The macular hole-free survival at 48 months was 94% for stage 0-negative patients, versus 54% for stage 0-positive patients. Univariate analysis revealed that the presence of a stage 0 macular hole was significantly associated with an almost 6-fold increase in the risk of macular hole formation (relative risk: 5.8, 95% confidence interval: 1.16-28.61, P = 0.03). CONCLUSIONS: A stage 0 macular hole has a normal biomicroscopic appearance clinically, but has salient features on OCT as a result of oblique vitreous traction. Optical coherence tomographic findings consist of a normal foveal contour and normal retinal thickness and must include the presence of a preretinal, minimally reflective, thin band inserting obliquely on at least one side of the fovea. The presence of a stage 0 macular hole in the fellow eye is a significant risk factor for the development of a second macular hole. PMID- 15522369 TI - Comparison of ultrahigh- and standard-resolution optical coherence tomography for imaging macular hole pathology and repair. AB - PURPOSE: To compare ultrahigh-resolution optical coherence tomography (UHR-OCT) technology to a standard-resolution OCT instrument for the imaging of macular hole pathology and repair; to identify situations where UHR-OCT provides additional information on disease morphology, pathogenesis, and management; and to use UHR-OCT as a baseline for improving the interpretation of the standard resolution images. DESIGN: Observational and interventional case series. PARTICIPANTS: Twenty-nine eyes of 24 patients clinically diagnosed with macular hole in at least one eye. METHODS: A UHR-OCT system has been developed and employed in a tertiary-care ophthalmology clinic. Using a femtosecond laser as the low-coherence light source, this new UHR-OCT system can achieve an unprecedented 3-mum axial resolution for retinal OCT imaging. Comparative imaging was performed with UHR-OCT and standard 10-mum resolution OCT in 29 eyes of 24 patients with various stages of macular holes. Imaging was also performed on a subset of the population before and after macular hole surgery. MAIN OUTCOME MEASURES: Ultrahigh- and standard-resolution cross-sectional OCT images of macular hole pathologies. RESULTS: Both UHR-OCT and standard-resolution OCT exhibited comparable performance in differentiating various stages of macular holes. The UHR-OCT provided improved imaging of finer intraretinal structures, such as the external limiting membrane and photoreceptor inner segment (IS) and outer segment (OS), and identification of the anatomy of successful surgical repair. The improved resolution of UHR-OCT enabled imaging of previously unidentified changes in photoreceptor morphology associated with macular hole pathology and postoperative repair. Visualization of the junction between the photoreceptor IS and OS was found to be an important indicator of photoreceptor integrity for both standard-resolution and UHR-OCT images. CONCLUSIONS: Ultrahigh resolution optical coherence tomography improves the visualization of the macular hole architectural morphology. The increased resolution of UHR-OCT enables the visualization of photoreceptor morphology associated with macular holes. This promises to lead to a better understanding of the pathogenesis of macular holes, the causes of visual loss secondary to macular holes, the timing of surgical repair, and the evaluation of postsurgical outcome. Ultrahigh-resolution optical coherence tomography imaging of macular holes that correspond to known alterations in retinal morphology can be used to interpret retinal morphology in UHR-OCT images. Comparisons of UHR-OCT images with standard-resolution OCT images can establish a baseline for the better interpretation of clinical standard resolution OCT images. The ability to visualize photoreceptors and their integrity or impairment is an indicator of macular hole progression and surgical outcome. PMID- 15522370 TI - Intravitreal triamcinolone for diabetic macular edema that persists after laser treatment: three-month efficacy and safety results of a prospective, randomized, double-masked, placebo-controlled clinical trial. AB - OBJECTIVE: To determine whether an intravitreal injection of triamcinolone acetonide for persistent diabetic macular edema after adequate laser treatment improves visual acuity. DESIGN: Prospective, double-masked, placebo-controlled, randomized clinical trial. PARTICIPANTS AND CONTROLS: Sixty-nine eyes of 43 patients were entered into the study, with 34 eyes randomized to receive active treatment and 35 randomized to receive a placebo injection. Sixty-five of 69 eyes (94%) completed the 3-month study visit. INTERVENTION: Using a 27-gauge needle, 0.1 ml of triamcinolone acetonide was injected through the pars plana. The procedure was performed in a minor procedures area in the outpatient clinic under sterile conditions and using topical and subconjunctival anesthesia. Eyes randomized to placebo received a subconjunctival saline injection using the identical procedure for preparation. MAIN OUTCOME MEASURES: The main outcome measures were improvement of best-corrected logarithm of the minimum angle of resolution visual acuity by 5 or more letters and incidence of moderate or severe adverse events. RESULTS: Eighteen of 33 eyes (55%) treated with triamcinolone gained 5 or more letters of best-corrected visual acuity compared with 5 of 32 eyes (16%) treated with placebo (P = 0.002). Macular edema was reduced by 1 or more grades as determined by masked semiquantitative contact lens examination in 25 of 33 treated eyes (75%) versus 5 of 32 untreated eyes (16%; P<0.0001). Optical coherence tomography showed a mean reduction of central retinal thickness of 152 mum in the 21 treated eyes that were examined compared with a reduction of 36 mum in 20 placebo-treated eyes. Infectious endophthalmitis developed in 1 triamcinolone-treated eye that was treated adequately without loss of visual acuity. CONCLUSIONS: In the short term, intravitreal triamcinolone is an effective and relatively safe treatment for eyes with diabetic macular edema that have failed laser treatment. Although it will be essential to study longer-term outcomes, the use of intravitreal triamcinolone may be considered in 1 eye of patients who continue to lose vision from diabetic macular edema despite conventional management. PMID- 15522371 TI - Delayed absorption of macular edema accompanying serous retinal detachment after grid laser treatment in patients with branch retinal vein occlusion. AB - PURPOSE: To study the detailed process of macular edema (ME) absorption after grid laser photocoagulation in patients with branch retinal vein occlusion (BRVO). The influence of pretreatment serous retinal detachment (SRD) at the fovea on patient outcome was also evaluated. DESIGN: Retrospective, interventional, comparative case series. PARTICIPANTS: Thirty-seven BRVO patients presenting with severe ME. METHODS: Patients were treated with grid laser photocoagulation with 6 months of follow-up examinations. Baseline and post treatment examinations included measurements of visual acuity (VA), fluorescein angiography, and detailed imaging of ME by optical coherence tomography (OCT). Macular thickness was defined as the distance from the inner retinal surface to the outer border of the sensory retina (foveal retinal thickness) and also to the inner border of the retinal pigment epithelium including the SRD (total foveal elevation). MAIN OUTCOME MEASURES: Post-treatment macular thickness with OCT, VA converted to the logarithm of the minimum angle of resolution, and absorption of SRD, and correlations of macular thickness and VA both before and after treatment. RESULTS: Total foveal elevation and VA were significantly improved at 1, 3, and 6 months after treatment. There was also a significant correlation between reduction of total foveal elevation and increase in VA. Fourteen eyes (37.8%) displayed SRD, as evidenced by OCT at the baseline. In eyes without SRD, foveal retinal thickness and VA had significantly improved in a time-dependent manner. However, although SRD itself was almost absorbed 6 months after treatment, the improvement of both VA and total foveal elevation in eyes with pretreatment SRD was not significant compared with baseline conditions. Both post treatment total foveal elevation and VA of eyes with SRD tended to be worse than eyes without SRD. CONCLUSION: The presence of subfoveal SRD retards the absorption of ME and recovery of VA after grid laser photocoagulation in patients with BRVO. PMID- 15522372 TI - Ocular manifestations of dengue fever. AB - PURPOSE: To evaluate ocular manifestations associated with dengue fever. DESIGN: Retrospective case series and literature review. METHODS: Clinical records of patients with dengue fever who subsequently had ocular symptoms and signs develop were reviewed. The clinical presentation and ocular complications were evaluated. RESULTS: Six patients, 5 females and 1 male, were seen with a sudden decrease in vision 6 to 7 days after the initial manifestations of dengue fever were identified. The diagnosis was confirmed by detection of dengue-specific IgM antibodies (IgM enzyme-linked immunoassay). The presenting best-corrected visual acuity ranged from 20/30 to counting fingers, and ocular involvement was bilateral but asymmetric in 5 cases and unilateral in 1 case. Fundus findings included small, intraretinal, whitish lesions, with localized retinal and retinal pigment epithelium (RPE) disturbance, small dot hemorrhages, and vascular sheathing around the macula and the papillomacular bundle. Fluorescein angiography showed arteriolar focal knobby hyperfluorescence at the macula with mild staining of the vascular walls and leakage at the level of the RPE. All 5 cases that had indocyanine green angiography done showed early diffuse choroidal hyperfluorescence with late silhouetting of the larger choroidal vessels. Five patients received steroid therapy: 1 topical, 2 periocular, and 2 oral. Over 2 to 4 months, RPE discoloration was observed over the affected areas. After the acute episode, 3 patients showed partial recovery of vision, and in the remaining patients, the visual acuity remained stable. CONCLUSIONS: Ocular complications associated with dengue fever are rare but may result in permanent visual impairment. PMID- 15522373 TI - Chorioretinal involvement in patients with West Nile virus infection. AB - PURPOSE: To characterize and analyze the chorioretinal manifestations of West Nile virus (WNV) infection. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Twenty-nine consecutive patients with serologically confirmed WNV infection in the setting of an outbreak of the disease. METHODS: The average duration of systemic symptoms before ophthalmic examination was 10 days (range, 2 30 days). All participants underwent a detailed ophthalmic examination at presentation and regularly throughout follow-up, including dilated biomicroscopic fundus examination, fundus photography, and fluorescein angiography. RESULTS: A typical multifocal chorioretinitis was observed in 20 of 29 patients (69%) bilaterally at presentation and developed later during follow-up in 3 patients (10.3%), bilaterally (n = 1) or unilaterally (n = 2). Multifocal chorioretinitis was associated with mild vitreous inflammatory reaction in all cases. Other findings included intraretinal hemorrhages (21 patients [72.4%]), white-centered retinal hemorrhages (7 patients [24.1%]), focal retinal vascular sheathing (3 patients [10.4%]), marked diffuse retinal arterial sheathing (1 patient [3.4%]), retinal vascular leakage (5 patients [17.2%]), optic disc swelling (2 patients [6.9%]), optic disc staining (6 patients [20.7%]), segmental zones of retinal pigment epithelium changes (1 patient [3.4%]), and nonproliferative diabetic retinopathy (7 patients [24.1%]). The posterior segment findings related to WNV disease had a self-limited course in all patients. CONCLUSIONS: Chorioretinal involvement, frequently asymptomatic and self-limited, is common in patients with acute WNV infection. The unique pattern of multifocal chorioretinitis in patients with systemic symptoms suggestive of WNV can help to establish the diagnosis while serologic testing is pending. Therefore, a systematic ocular evaluation, including dilated fundus examination and fluorescein angiography in selected cases, is recommended in patients with clinically suspected WNV infection. PMID- 15522374 TI - Lack of herpes virus DNA in choroidal tissues of a patient with serpiginous choroiditis. AB - OBJECTIVE: To investigate the role of herpes viruses in the etiology of serpiginous choroiditis. DESIGN: Interventional case report. PARTICIPANT: A 59 year-old male patient with long-term history of serpiginous choroiditis. INTERVENTION: The patient's affected eye was obtained during autopsy. Polymerase chain reaction was performed in the microdissected choroidal tissues. RESULTS: Histopathologic examination demonstrated active inflammation with lymphocytic infiltration of the choroid. No viral DNA was amplified using pairs of herpes simplex virus (HSV) P1/P2 (for HSV-1, HSV-2, Epstein-Barr virus [EBV], cytomegalovirus [CMV] and human herpes virus [HHV]-8), and varicella-zoster virus [VZV] P1/P2 (for VZV, HHV-6, HHV-7) in the infiltrating lymphocytes or choroidal tissues. CONCLUSIONS: The current observation suggests a lack of a role for herpetic viral etiology in the etiopathogenesis of serpiginous choroiditis. PMID- 15522375 TI - A randomized trial of atropine regimens for treatment of moderate amblyopia in children. AB - OBJECTIVE: To compare daily atropine to weekend atropine as prescribed treatments for moderate amblyopia in children younger than 7 years. DESIGN: Prospective, randomized multicenter clinical trial (30 sites). PARTICIPANTS: One hundred sixty eight children younger than 7 years with amblyopia in the range of 20/40 to 20/80 associated with strabismus, anisometropia, or both. INTERVENTION: Randomization either to daily atropine or to weekend atropine for 4 months. Partial responders were continued on the randomized treatment until no further improvement was noted. MAIN OUTCOME MEASURE: Visual acuity (VA) in the amblyopic eye after 4 months. RESULTS: The improvement in VA of the amblyopic eye from baseline to 4 months averaged 2.3 lines in each group. The VA of the amblyopic eye at study completion was either (1) at least 20/25 or (2) better than or equal to that of the sound eye in 39 children (47%) in the daily group and 45 children (53%) in the weekend group. The VA of the sound eye at the end of follow-up was reduced by 2 lines in one patient in each group. Stereoacuity outcomes were similar in the 2 groups. CONCLUSIONS: Weekend atropine provides an improvement in VA of a magnitude similar to that of the improvement provided by daily atropine in treating moderate amblyopia in children 3 to 7 years old. This article contains additional online-only material available at http://www.ophsource.com/periodicals/ophtha. PMID- 15522376 TI - Ocular complications in the Department of Defense Smallpox Vaccination Program. AB - OBJECTIVE: The purpose of this case series was to present an overview of the nature and frequency of ocular complications in the Department of Defense (DoD) Smallpox Vaccination Program. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: The authors retrospectively evaluated data collected on individuals with an ophthalmologic complaint after receiving smallpox vaccination or after contact with a recently immunized individual. The vaccinee and contact cases occurred secondary to inoculations given between December 13, 2002 and May 28, 2003 as part of the DoD Smallpox Vaccination Program. METHODS: Data were collected primarily from reports to military headquarters or to the Vaccine Adverse Event Reporting System and individual medical records. MAIN OUTCOME MEASURES: The incidence, types, and timing of ocular complications were evaluated. Diagnostic and treatment considerations also were reviewed. RESULTS: Between December 13, 2002 and May 28, 2003, 450,293 smallpox vaccinations were given. We identified 16 confirmed or probable cases of ocular vaccinia, with an incidence of 3.6 per 100,000 inoculations. Of these cases, 12 (75%) were seen in the vaccinees, and 4 (25%) in close contacts. Of the 12 self-inoculation cases, 7 (58.3%) were seen in individuals receiving the vaccine for the first time (primary vaccination), and 3 (25.0%) were seen in individuals previously vaccinated (revaccination); the vaccination status in 2 cases was unknown. Clinical manifestations included lid pustules, blepharitis, periorbital cellulitis, conjunctivitis, conjunctival ulcers, conjunctival membranes, limbal pustules, corneal infiltrates, and iritis, with onset of symptoms 3 to 24 days after inoculation or contact. Five of 9 tested cases were culture or polymerase chain reaction positive for vaccinia. Treatment for most cases was topical trifluridine 1% (Viroptic; King Pharmaceuticals, Inc., Bristol, TN). Vaccinia immune globulin was used in 1 case. In all patients, recovery occurred without significant visual sequelae. CONCLUSIONS: When compared with historical data on the ocular complications of smallpox vaccination, the incidence of ocular complications during the DoD Smallpox Vaccination program has been low. In addition, the severity of disease seems to be less than during other vaccination periods. These findings perhaps are the result of improved screening of vaccinees, prevaccination counseling, postvaccination wound care, and the suggested efficacy of trifluridine in the treatment of ocular vaccinia. PMID- 15522377 TI - Late postoperative opacification of a hydrophilic acrylic (hydrogel) intraocular lens: a clinicopathological analysis of 106 explants. AB - PURPOSE: To report clinical, pathologic, histochemical, ultrastructural, and spectroscopic analyses of MemoryLens intraocular lenses (IOLs) explanted from patients who had visual disturbances caused by postoperative opacification of the lens optic. DESIGN: Noncomparative, large case series with clinicopathologic correlation. PARTICIPANTS: A total of 106 hydrophilic acrylic IOLs of the same design explanted from 106 different patients. All patients had decreased visual acuity at presentation approximately 2 years after cataract surgery, associated with a whitish fine granularity on the optical surfaces of the IOLs. METHODS: The explanted IOLs were submitted to the John A. Moran Eye Center and were examined under light microscopy, histochemically, and with scanning electron microscopy (SEM) equipped with an energy dispersive x-ray spectroscopy detector with light element capabilities (EDS). MAIN OUTCOME MEASURES: The IOLs were examined for distribution, structure, and composition of the deposits causing opacification of their optic components. RESULTS: The average interval between lens implantation and opacification was 25.8+/-11.9 months. The most frequently associated medical and ophthalmic conditions were diabetes and glaucoma. However, some patients did not have any preexisting medical or ophthalmic conditions. Most of the IOLs had been implanted in 1999 and 2000. Microscopic analyses revealed the presence of multiple fine, granular deposits of variable sizes on the anterior and posterior optic surfaces, especially on the anterior surface. The deposits stained positive for calcium. The EDS confirmed the presence of calcium and phosphate within the deposits. CONCLUSIONS: The results obtained suggest the surface deposits to be composed, at least in part, by calcium and phosphate. A special polishing technique used in the manufacture of most of these IOLs may have caused changes in the lens surface leading to deposit formation. Further studies should be undertaken to confirm this hypothesis. PMID- 15522378 TI - A single intraoperative sub-Tenon's capsule triamcinolone acetonide injection for the treatment of post-cataract surgery inflammation. AB - PURPOSE: To compare a single intraoperative sub-Tenon's capsule triamcinolone acetonide injection with steroid drops in the treatment of ocular inflammation after cataract surgery. DESIGN: Randomized, double-masked controlled trial. PARTICIPANTS: A total of 100 patients were randomized prospectively into 2 groups: 50 patients treated with 1% prednisolone eyedrops (control group A) and 50 patients treated with sub-Tenon's capsule triamcinolone (treatment group B). METHODS: All patients underwent phacoemulsification and intraocular posterior lens implantation. After surgery, patients were randomized to receive either (group B) an intraoperative 40 mg triamcinolone acetonide sub-Tenon's capsule injection or (group A) 1% prednisolone acetate eyedrops, according to the following schedule: 1 drop 4 times daily (week 1), 3 times daily (week 2), 2 times daily (week 3), once daily (week 4). To mask the study, group B received vehicle drops administered on a similar schedule, and group A received an intraoperative sub-Tenon's capsule injection of a 1 ml balanced salt solution. MAIN OUTCOME MEASURES: The main outcome measures included inflammation (cell, flare, ciliary flush), intraocular pressure, and lack of response. RESULTS: Triamcinolone was shown to have anti-inflammatory efficacy clinically equivalent to conventional 1% prednisolone eyedrops in reducing intraocular inflammation, as measured by clinical methods. Triamcinolone was found to be as safe as the prednisolone in terms of adverse effects, changes in visual acuity, intraocular pressure, and biomicroscopic and ophthalmoscopic variables. On the third, seventh, fourteenth, and twenty-eighth postoperative days, a significantly lower intraocular pressure (P<0.01) was noted in the triamcinolone group than in the prednisolone group. CONCLUSIONS: A single intraoperative 40-mg triamcinolone acetonide sub-Tenon's capsule injection demonstrated a clinically equivalent therapeutic response and ocular tolerance compared with 1% prednisolone drops in controlling postoperative inflammation after uncomplicated cataract surgery and merits further investigation. PMID- 15522379 TI - The Advanced Glaucoma Intervention Study (AGIS): 14. Distinguishing progression of glaucoma from visual field fluctuations. AB - PURPOSE: To determine the least worsening of a visual field (VF) and the least number of confirming tests needed to identify progression of glaucomatous VF defects. DESIGN: Cohort study of participants in a clinical trial. PARTICIPANTS: Seven hundred fifty-two eyes of 565 patients with advanced glaucoma. METHODS: Visual field tests were quantified with the Advanced Glaucoma Intervention Study (AGIS) VF defect score and the Humphrey Field Analyzer mean deviation (MD). Follow-up was 8 to 13 years. MAIN OUTCOME MEASURES: Two measures based on the AGIS VF defect score: (1) sustained decrease of VF (SDVF), a worsening from baseline by 2 (alternatively, 3 or 4) or more units and sustained for 2 (alternatively, 3) consecutive 6-month visits and (2) after the occurrence of SDVF, the average percent of eyes with worsening by 2 (alternatively, 3 or 4) or more units from baseline. Two similar measures based on MD. RESULTS: Based on the original AGIS criteria for SDVF (a worsening of 4 units in the AGIS score sustained during 3 consecutive 6-month visits), 31% of eyes had an SDVF. The percent of eyes with a sustained event increases by approximately 10% when either the minimum number of units of field loss or the minimum number of 6-month visits during which the loss is sustained decreases by 1. During 3 years of follow-up after a sustained event, a worsening of at least 2 units was found in 72% of eyes that had a 2-visit sustained event. The same worsening was found in 84% of eyes that had a 3-visit sustained event. Through the next 10 years after a sustained event, based on worsening of 2, 3, or 4 units at 2 or 3 consecutive tests, the loss reoccurred, on average, in >/=75% of study eyes. Results for MD are similar. CONCLUSIONS: In patients with advanced glaucoma, a single confirmatory test 6 months after a VF worsening indicates with at least 72% probability a persistent defect when the worsening is defined by at least 2 units of AGIS score or by at least 2 decibels of MD. When the number of confirmatory tests is increased from 1 to 2, the percentage of eyes that show a persistent defect increases from 72% to 84%. PMID- 15522380 TI - Statistical evaluation of the diagnostic accuracy of methods used to determine the progression of visual field defects in glaucoma. AB - OBJECTIVE: To evaluate trend-type analyses to judge the progression of visual field damage (VFD) of glaucoma in terms of ratios of making judgments of progression and specificity, and to report a new method. DESIGN: Retrospective analysis of visual field (VF) results of actual glaucoma cases and those simulated by computer, and receiver operating characteristic curve analysis of performance of methods to judge the progression of VFD. PARTICIPANTS: One hundred five eyes of 105 open-angle glaucoma (OAG) patients with progressing VFD and 355 eyes of 355 clinically stable OAG patients for VF simulation from 4 university based referral practices. METHODS: Methods using regression analysis of total deviation (TD), mean deviation (MD), mean TD of a sectored VF, and original scoring used in the Advanced Glaucoma Intervention Study (AGIS) were compared. A VF test was repeated twice in a short period on the 355 stable OAG eyes, and test retest fluctuation, including variance at each test point and covariance between 2 test points, was calculated to simulate stable glaucomatous VF series by computer. The sensitivity of each method was calculated with 105 progressing VF series, and specificity was calculated with 10,000 simulated stable glaucomatous VF series. MAIN OUTCOME MEASURES: Sensitivity (ratios of making judgments of progression), specificity, and diagnostic power. RESULTS: The methods using the TD slope on one test location showed a sensitivity of 0.848 to 1.000, with a specificity of 0.105 to 0.721, and on 2 adjacent test locations showed a sensitivity of 0.848, with a specificity of 0.722. A significant negative MD slope with P<0.05 showed a sensitivity of 0.524, with a specificity of 0.945. The method using a sectored VF showed a sensitivity of 0.695, with a high specificity of 0.946. The AGIS method showed a sensitivity of 0.305 to 0.467, with a very high specificity of 0.999 to 1.000. The method using previously reported mathematically sectored VFs showed a sensitivity of 0.790, with a specificity of 0.900, and higher diagnostic power (1.69) than the others in this study population. CONCLUSIONS: Most of the methods using the TD slope were characterized by high sensitivity, the AGIS method had a very high specificity, and those using VF sectors had reasonable sensitivity and specificity. PMID- 15522381 TI - Central corneal pachymetry and visual field progression in patients with open angle glaucoma. AB - PURPOSE: To investigate the association between corneal pachymetry and visual field progression in patients with chronic open-angle glaucoma. DESIGN: Retrospective case-control study. PARTICIPANTS: Eighty-eight patients with primary open-angle glaucoma, pseudoexfoliative glaucoma, pigmentary glaucoma, or normal-tension glaucoma, followed for an average of 8 years, who had visual field loss and progression as defined by modified Anderson criteria. Cases with progression were matched for race, diagnosis, and age at pachymetry with controls who did not have progression. METHODS: Progression was defined by use of the modified Anderson criteria. Central corneal thickness (CCT) was determined by ultrasound pachymetry. MAIN OUTCOME MEASURES: Visual field progression and corneal pachymetry. RESULTS: The mean CCT in patients with visual field progression was significantly lower than the mean CCT in patients who did not progress (529+/-36 mum vs. 547+/-35 mum; P = 0.02). Those with thinner CCT were more likely to progress than those with thicker CCT as identified by Cox proportional hazards regression analysis (P = 0.01; hazard ratio, 1.44 for a 40 mum thinner CCT; 95% confidence interval, 1.12-1.80), and CCT was the only risk factor identified to be significantly associated with visual field progression. CONCLUSIONS: In this case-control patient population, visual field progression in patients with open-angle glaucoma was significantly associated with thinner CCT. PMID- 15522382 TI - Impact of diabetes on glaucoma screening using frequency-doubling perimetry. AB - PURPOSE: To determine whether diabetes is a potential source of abnormal test results in glaucoma screening by use of frequency-doubling perimetry. DESIGN: Observational case series. PARTICIPANTS AND CONTROLS: Twenty-five patients with diabetes and 25 nondiabetic normal controls, all without cataract or glaucoma. METHODS: Frequency-doubling perimetry using the C-20-5 screening mode was performed in 1 randomly selected eye of each participant. Abnormal test results were immediately repeated to reduce artifacts caused by test inexperience. Frequency-doubling technology (FDT) results were assigned a numerical score between 0 (normal) and 87 (no stimuli detected). MAIN OUTCOME MEASURE: Abnormal FDT test scores. RESULTS: Overall, 7 of 25 (28%) patients with diabetes and 1 of 25 (4%) normal controls had an abnormal test; the 1 normal subject with an abnormal test was subsequently determined to have an abnormal glucose tolerance test. Four of the 7 patients with diabetes with abnormal FDT tests had no evidence of diabetic retinopathy on clinical examination. Patients with diabetes had higher test scores than normal controls (8.9+/-16.9 vs. 1.0+/-4.4, P = 0.028) and had longer test times than normal controls (66.4+/-35.5 seconds vs. 48.3+/ 11.2 seconds, P = 0.019). CONCLUSIONS: Frequency-doubling perimetry is abnormal in some patients with diabetes, including some patients with diabetes without clinical evidence of diabetic retinopathy. Abnormal FDT testing in diabetic eyes may not represent glaucomatous visual field loss. Diabetes may be a source of "false-positive" test results when this technology is used for glaucoma screening. PMID- 15522384 TI - Silicone oil complications. PMID- 15522383 TI - Contact transscleral neodymium:yttrium-aluminum-garnet laser cyclophotocoagulation Long-term outcome. AB - PURPOSE: Contact transscleral neodymium:yttrium-aluminum-garnet (Nd:YAG) laser cyclophotocoagulation (CYC) is a treatment option for advanced glaucoma refractory to alternative treatments. This study determined the long-term efficacy and risks of contact transscleral Nd:YAG laser CYC. DESIGN: A prospective study was performed with patients with advanced, uncontrolled glaucoma who received CYC from 1988 through 1989. PARTICIPANTS: Records for 68 eyes of 64 patients were obtained and reviewed for the 10-year follow-up. METHODS: A transscleral continuous-wave Nd:YAG laser was used for photocoagulation of the ciliary body. MAIN OUTCOME MEASURES: Intraocular pressure (IOP), visual acuity, and second intervention. Failure was defined as the need for second intervention, IOP of more than 25 mmHg, or IOP of less than 3 mmHg. RESULTS: The mean follow-up period was 5.85+/-4.0 years (range, 0.1-10 years). The mean preoperative IOP of 36.3+/-10.1 mmHg decreased to 22.6+/-11.3 mmHg at 1 year of follow-up (P<0.001). The mean postoperative IOP at 5 years was 21.8+/ 13.3 mmHg (P<0.001) and was 18.9+/-12.2 mmHg at 10 years of follow-up (P<0.001). A second intervention after CYC was required in 30 eyes (44.1%). Six eyes (8.8%) with initial visual acuity of counting fingers or worse progressed to no light perception, and 5 of 8 eyes (62.5%) with visual acuity better than 20/200 lost 2 or more Snellen lines. Hypotony developed in 3 eyes (4.4%). Overall, the failure rate by 10 years of follow-up was 51.5% (35/68 eyes). CONCLUSIONS: Cyclophotocoagulation resulted in a significant reduction of IOP after surgery at 1, 5, and 10 years of follow-up; however, 51.5% of eyes failed by the end of 10 years, with most failures occurring within the first year (40%). Although CYC provides a useful method to lower IOP significantly, this study suggests that its success in controlling IOP is tempered by its failure rate and risk of complications, including visual loss, phthisis, and loss of light perception. PMID- 15522385 TI - Verteporfin in Photodynamic Therapy: report no. 5. PMID- 15522386 TI - Adhesion abnormalities associated with LASIK. PMID- 15522388 TI - The Australian Mohs database. PMID- 15522390 TI - Surgery for trichiasis. PMID- 15522393 TI - Copper(II) complexes of some N-substituted bis(aminomethyl)phosphinate ligands. An integrated EPR study of microspeciation and coordination modes by the two dimensional simulation method. AB - Copper(II) complexes of bis(aminomethyl)phosphinic acid (L1), bis(N-glycino-N methyl)phosphinic acid (L2), bis(N-benzylglycino-N-methyl)phosphinic acid (L3), bis(l-prolino-N-methyl)phosphinic acid (L4) and bis(iminodicarboxymethyl-N methyl)phosphinic acid (L5) were studied in aqueous solution by pH-potentiometric and electron paramagnetic resonance (EPR) spectroscopic methods. The EPR spectrum packages recorded at various ligand-to-metal concentration ratios and pH's were analyzed (after matrix rank analysis by the method of residual intensities as a complementary method) by the two-dimensional computer simulation method, which simultaneously determines the formation constants and the EPR parameters of the various (micro)species. L1 forms mono and bis complexes in different protonation states; for the other ligands, the mono complexes are always prevalent. For steric reasons, the formation of CuL is shifted to increasingly higher pH regions in the sequence L2, L3 and L4. CuLH was identified for L3, L4 and L5, and also CuLH(2) for L4 and L5. Cu(2)L(2) was found in small amounts for L3 and L4, while it predominates at pH>4 for L5. For L5, Cu(2)L(2)H(2) was also detected. For the ligands that form dimeric metal complexes in equimolar solution or at a ligand excess, Cu(2)L is formed at a metal ion excess. Ligation of the phosphinate O was suggested by indirect proofs in the protonated complexes of L1. For the ligands L2, L3 and L4, the copper(II) coordination in various species in different protonation states is reminiscent of that in the mono and bis complexes of simple amino acids. For the bis(aminomethyl)phosphinates, however, the cis positions of the amino groups in CuL are ensured by the structure of the ligand, and the isomers differ from each other in the (equatorial or axial) position of the second carboxylate group. PMID- 15522394 TI - Symphoria: the success of modeling the active site function of oxo molybdoenzymes. AB - The success of modeling the active site function of oxomolybdoenzymes have been claimed generally on the basis of reactivity of the synthetic analogues towards PPh(3) or DMSO (dimethyl sulfoxide). Here it has been shown that the success of modeling the active site function of these enzymes may not be determined by the ability of a model to undergo oxotransfer with PPh(3) or DMSO (except for the modeling of DMSO reductase) and one should adhere to the criteria accepted by the bioinorganic community. A critical evaluation of two of those criteria which requires a synthetic analogue (a) should react with the enzyme substrate (b) should follow the same rate law as does the enzyme, has been presented in this paper. We have shown that the fulfillment of criterion (b) and the inhibition phenomena to that effect both are dictated by symphoria (from sympherin in Greek: the bringing together of reactants into the proper spatial relationship) on the basis of kinetic studies of the reactivity of enzyme substrate the HSO(3)(-) and its analogues (anions of oxyacids of phosphorous) towards a functional model sulfite oxidase [Bu(4)N](2)[Mo(VI)O(2)(mnt)(2)] (mnt(2-)=1,2 dicyanoethylenedithiolate) but with the caveat that the mechanistic inference drawn from such studies may not be the same as in the case of native enzyme. In view of this ambiguity it has been pointed out that the fulfillment of this criterion is not a definitive conclusion towards our understanding of the structure-function relationship of an enzyme and, therefore, the criterion of a 'structural analogue' and 'functional analogue' have been revised subject to an amendment of criterion (a) to include substrate analogues. It has also been shown for the first time on the basis of kinetic studies that the effect of medium can lead to substrate - inhibitor type dualism and hence the effect of medium is also a factor that can play a key role for the success of modeling the active site function of an enzyme. Here we also provide the details of the inhibition mechanisms proposed in our earlier report with an indirect proof to that effect. PMID- 15522395 TI - Inhibiting effect of selenium on oxysterols-induced apoptosis of rat vascular smooth muscle cells. AB - To evaluate the cytoprotection mechanism of selenium against cholestane 3beta,5alpha,6beta-triol (3-triol)-induced vascular smooth muscle cells (VSMCs) damage, cell viability was analyzed by 3-(4,5-dimethylthiazol-2 -yl)-2,5 diphenyltetrazolium bromide (MTT) assay and cell count, the percentage release of lactate dehydrogenase (LDH) from the cell was assessed, and apoptosis was detected by DNA laddering and flow cytometric analysis. Meanwhile, the activity of glutathione peroxidase (GPx) of VSMCs was measured. The results showed that 3 triol could inhibit proliferation of VSMCs time-dependently and dose-dependently, increase the percentage release of LDH and induce VSMCs apoptosis. While the cytotoxicity and cells apoptosis induced by 3-triol was attenuated by pretreatment of cells with low concentration of sodium selenite, and the longer the pretreated time was, the stronger the inhibition was. Preincubation of cells with sodium selenite (50 nM) for 12 or 24 h before 1, 5, 10, 25, or 50 microM 3 triol exposure, the cell viabilities increased 28.5% (P<0.05), 18.3%, 197.6% (P<0.01), 66.7%, 50.0% or 35.1% (P<0.05), 62.3% (P<0.05), 329.6% (P<0.01), 221.3% (P<0.05), 74.0% compared with the control cells, respectively. When the cells were preincubated with sodium selenite (50 nM) for 12 or 24 h before exposure to 3-triol (10 microM), the percent of apoptotic cells reduced from 30.47+/-15.34% to 26.88+/-17.32% or 7.41+/-5.46% (P<0.05). With preincubation of sodium selenite (50 nM) for 24 h, the GPx activity of VSMCs increased 18.5% compared with control (P<0.05). In conclusion, the results suggested that incubated VSMCs could absorb and transfer selenite as selenoprotrein, such as GPx, if the time is long enough and VSMCs selenoproteins can protect markedly against apoptosis and damage induced by 3-triol in VSMCs. PMID- 15522396 TI - Crystal structures of ferrous and ferrous-NO forms of verdoheme in a complex with human heme oxygenase-1: catalytic implications for heme cleavage. AB - Heme oxygenase oxidatively degrades heme to biliverdin resulting in the release of iron and CO through a process in which the heme participates both as a cofactor and substrate. One of the least understood steps in the heme degradation pathway is the conversion of verdoheme to biliverdin. In order to obtain a better understanding of this step we report the crystal structures of ferrous-verdoheme and, as a mimic for the oxy-verdoheme complex, ferrous-NO verdoheme in a complex with human HO-1 at 2.20 and 2.10 A, respectively. In both structures the verdoheme occupies the same binding location as heme in heme-HO-1, but rather than being ruffled verdoheme in both sets of structures is flat. Both structures are similar to their heme counterparts except for the distal helix and heme pocket solvent structure. In the ferrous-verdoheme structure the distal helix moves closer to the verdoheme, thus tightening the active site. NO binds to verdoheme in a similar bent conformation to that found in heme-HO-1. The bend angle in the verodoheme-NO structure places the terminal NO oxygen 1 A closer to the alpha-meso oxygen of verdoheme compared to the alpha-meso carbon on the heme NO structure. A network of water molecules, which provide the required protons to activate the iron-oxy complex of heme-HO-1, is absent in both ferrous-verdoheme and the verdoheme-NO structure. PMID- 15522397 TI - Oxidative DNA damage induced by HEPES (2-[4-(2-hydroxyethyl)-1 piperazinyl]ethanesulfonic acid) buffer in the presence of Au(III). AB - Oxidative DNA damage was investigated by free radicals generated from HEPES (2-[4 (2-hydroxyethyl)-1-piperazinyl]ethanesulfonic acid) buffer, which is widely used in biochemical or biological studies, in the presence of Au(III). The effect of free radicals on the DNA damage was ascertained by gel electrophoresis, electron spin resonance (ESR) spectroscopy and circular dichroism (CD) spectroscopy. ESR results indicated the generation of nitrogen-centered cationic free radicals from the HEPES in the presence of Au(III) which cause the DNA damage. No ESR spectra were observed for phosphate, tris(hydroxymethyl)aminomethane (Tris-HCl) and acetate buffers in the presence of Au(III) or for HEPES buffer in the presence of other metal ions such as Mn(II), Fe(III), Co(II), Ni(II), Cu(II), Zn(II) and Pd(II) or [Au(III)(TMPyP)](5+) and [Pd(II)(TMPyP)](4+), where [H(2)(TMPyP)](4+) denotes tetrakis(1-methylpyridium-4-yl)porphyrin. Consequently, no DNA damage was observed for these buffer agents (e.g., phosphate, Tris-HCl or acetate) in the presence of Au(III) or for HEPES in the presence of other metal ions or the metalloporphyrins mentioned above. No detectable inhibitory effect on the DNA damage was observed by using the typical scavengers of reactive oxygen species (ROS) ()OH, O(2)(-) and H(2)O(2). This non-inhibitory effect indicated that no reactive oxygen species were generated during the incubation of DNA with HEPES and Au(III). The drastic change in CD spectra from positive ellipticity to negative ellipticity approximately at 270 nm with increasing concentration of Au(III) also indicated the significant damage of DNA. Only HEPES or Au(III) itself did not damage DNA. A mechanism for the damaging of DNA is proposed. PMID- 15522398 TI - Ternary complexes metal [Co(II), Ni(II), Cu(II) and Zn(II)]--ortho-iodohippurate (I-hip)--acyclovir. X-ray characterization of isostructural [(Co, Ni or Zn)(I hip)(2)(ACV)(H(2)O)(3)] with stacking as a recognition factor. AB - Four ternary metal--ortho-iodohippurate (I-hip)--acyclovir (ACV) complexes, [M(I hip)(2)(ACV)(H(2)O)(3)] where M is Co(II) (1), Ni(II) (2), Cu (3) and Zn(II) have been obtained by reaction between the corresponding binary complexes M(II)(I hip)(2)xnH(2)O and ACV. Three ternary complexes (M=Co, Ni and Zn) and the corresponding Zn(II)--ortho-iodohippurate binary derivative have been structurally characterized by X-ray diffraction: The studies show these three ternary complexes are isostructural and present, in solid state, an interesting stacking between the nucleobase and the aryl ring of the hippurate moiety, which probably promotes the formation of ternary complexes. Moreover, the two different ligands interact between them by means of ancillary hydrogen bonds with water molecules coordinated to the metal ion. It must be mentioned that these two recognition factors, hydrogen bonds plus stacking, could explain the reason for the isostructurality of these ternary derivatives with so different three metal ions, with diverses trends in coordination numbers and geometries. In solid state, there are two enantiomeric molecules that are related by an inversion center as the crystal-building unit (as a translational motif) for the ternary complexes. PMID- 15522399 TI - Biologically relevant tetraazamacrocyclic complexes of manganese: synthetic, spectral, antimicrobial, antifertility and antiinflammatory approach. AB - The testicular sperm density, sperm morphology, sperm motility, density of cauda epididymis spermatozoa and fertility in mating trials and biochemical parameters of reproductive organs have been examined and discussed for [Mn(N(4)MacL(n))(NO(3))(2)] (where N(4)MacL(n) represents the tetraazamacrocyclic ligand molecule with n=1-4) type of complexes. An attempt also has been made to correlate the structural aspects of the compounds with their anti-inflammatory activity. The 14-18 membered tetraamide macrocyclic ligands N(4)L(1)-N(4)L(4) used during these investigations have been prepared by the condensation of 1,2 diaminoethane or 1,3-diaminopropane with malonic or succinic acid in the presence of condensing reagents, dicyclohexylcarbodiimide and 4-dimethylaminopyridine. On reduction, these macrocyclic ligands give a new series of tetraazamacrocyles [N(4)MacL(n)] and their complexes with manganese(II) nitrate. The ligands and their complexes were characterized by elemental analyses, molecular weight determinations, infrared, electronic, mass and X-ray structural analyses. An octahedral geometry for these complexes has been confirmed by spectral studies. On the basis of the chemical composition, the representation of the complexes as [Mn(N(4)MacL(n))(NO(3))(2)] has been established. The ligands and their complexes have been screened in vitro against a number of pathogenic fungi and bacteria to assess their growth inhibiting potential. PMID- 15522400 TI - Studies on activities, cell up take and DNA binding of four multinuclear complexes of the form: [[trans-PtCl(NH(3))(2)](2)mu-[trans-Pd(NH(3))(2) (H(2)N(CH(2))(n)NH(2))(2)]]Cl(4) where n=4-7. AB - The activity against human cancer cell lines including ovarian: A2780, A2780(cisR), cell up take, DNA-binding and nature of interaction with pBR322 plasmid DNA have been studied for four multinuclear complexes code named DH4Cl, DH5Cl, DH6Cl and DH7Cl, having the general formula: [[trans-PtCl(NH(3))(2)](2)mu [trans-Pd(NH(3))(2)-(H(2)N(CH(2))(n)NH(2))(2)]]Cl(4) where n=4, 5, 6 and 7 for DH4Cl, DH5Cl, DH6Cl and DH7Cl, respectively. The compounds are found to exhibit significant anticancer activity against ovarian cancer cell lines: A2780, A2780(cisR) and A2780(ZD0473R). DH6Cl in which the linking diamine has six carbon atoms is found to be the most active compound. As the number of carbon atoms in the linking diamine is decreased below six and increased above six, the activity is found to decrease, illustrating structure-activity relationship. All the multinuclear compounds are believed to form a plethora of long-range interstrand GG adducts with DNA dictated by the sequence of bases in the DNA strands. Increasing prevention of BamH1 digestion with the increase in concentration of the compounds is due to global changes in DNA conformation brought about by interstrand long-range binding of the compounds with DNA. PMID- 15522401 TI - Synthesis, crystal structures, and anti-convulsant activities of ternary [Zn(II)(3,5-diisopropylsalicylate)(2)], [Zn(II)(salicylate)(2)] and [Zn(II)(aspirinate)(2)] complexes. AB - Following observations that bis(3,5-diisopropylsalicylato)diaquazinc(II), [Zn(II)(3,5-DIPS)(2)(H(2)O)(2)], had anti-convulsant activity, bis(acetylsalicylate)diaquazinc(II), [Zn(II)(aspirinate)(2)(H(2)O)(2)], and the Zn(II) ternary 1,10-phenanthroline (phen), 2,9-dimethyl-1,10-phenanthroline (neocuproine, NC) or dimethyl sulfoxide (DMSO) complexes of Zn(II)3,5 diisopropylsalicylate, salicylate, and acetylsalicylate were synthesized and spectroscopically characterized. Anti-convulsant and Rotorod toxicity activities of these complexes were determined to examine their anti-convulsant and undesirable central nervous stimulant or depressant activities of these Zn(II) non-steroidal anti-inflammatory agent complexes. Bis(3,5-diisopropylsalicylato) 1,10-phenanthorlinezinc(II), [Zn(II)(3,5-DIPS)(2)(phen)], (1) has one bidentate phen ligand and two mono-deprotonated 3,5-DIPS ligands. One of the carboxylates bonds in an asymmetric chelating mode. The Zn(II) atom exhibits a distorted bicapped rectangular pyramidal environment N(2)O(2)OO (4+1+1 *). In the neocuproine complex, bis(3,5-diisopropylsalicylato)-2,9-dimethyl-1,10 phenanthorlinezinc(II), [Zn(II)(3,5-DIPS)(2)(NC)] (2), the Zn(II) atom has a much more distorted bicapped rectangular pyramidal environment, N(2)O(2)O(2) (4+2 *), compared to 1. The two carboxylate ligands exhibit the same asymmetric coordinating mode with longer metalloelement-oxygen bond distances compared to 1. The space group of [Zn(II)(aspirinate)(2)(H(2)O)(2)] (3), which has been reported as Cc is corrected to C2/c. The zinc atom exhibits a (4+2 *) bicapped square pyramidal environment. While the two ternary phenanthroline-containing complexes, 1 and 2, evidenced weak protection against maximal electroshock (MES)- and subcutaneous Metrazol (scMET) induced seizures, [Zn(II)(3,5-DIPS)(2)(DMSO)(2)], [Zn(II)(aspirinate)(2)(H(2)O)(2)], and bis(salicylato)-1,10 phenanthorlinezinc(II), [Zn(II)(salicylate)(2)(phen)], were found to be particularly useful in protecting against MES and scMET seizures and [Zn(II)(aspirinate)(2)(H(2)O)(2)] and [Zn(II)(salicylate)(2)(phen)] were found to have activity in protecting against Psychomotor seizures, without causing Rotorod toxicity. Activities of these and other Zn(II) complexes of non-steroidal anti inflammatory agents are consistent with the well-known anti-inflammatory responses of Zn(II)-dependent enzymes. There was also some evidence of Rotorod toxicity consistent with a mechanism of action involving sedative-hypnotic activity of recognized anti-epilepticdrugs. PMID- 15522402 TI - Direct photo-induced DNA strand scission by a ruthenium bipyridyl complex. AB - Irradiation of plasmid DNA in the presence of Ru(II)-2, a modified tris(2,2' bipyridyl)Ru(II) complex, in which two hydroxamic acid groups are attached to one of the three bipyridyl ligands, results in total fragmentation of the DNA. The photo-chemical reaction products were analyzed by gel electrophoresis, which revealed complete fragmentation. Further evidence for the complete degradation of the DNA was obtained by imaging the pre- and post-treated plasmid DNA using atomic force microscopy (AFM). A mechanism for the reaction is proposed. It initially involves the photo-chemical generation of Ru(III) ions and superoxide radicals, as corroborated by absorbance difference spectroscopy and electron paramagnetic resonance (EPR). Consequently, Ru(III) preferentially oxidizes guanine, liberating superoxide radicals that yield OH radicals. The OH radicals were identified by observing the spectral change at 532 nm of a 5'-dAdG substrate forming a colored adduct with thiobarbituric acid. These radicals are associated with the major non-specific damage exerted to DNA. PMID- 15522403 TI - Structure of carbohydrate-bound polynuclear iron oxyhydroxide nanoparticles in parenteral formulations. AB - Intravenous iron therapy is used to treat anemia associated with chronic kidney disease. The chemical structures of parenteral iron agents have not been characterized in detail, and correlations between structure, efficiency of iron delivery, and toxicity via catalysis of oxygen-derived free radical creation remain to be established. In this study, two formulations of parenteral iron have been characterized by absorption spectroscopy, X-ray diffraction analysis (XRD), transmission electron microscopy (TEM), atomic force microscopy (AFM), and elemental analysis. The samples studied were Venofer (Iron Sucrose Injection, USP) and Ferrlecit (Sodium Ferric Gluconate in Sucrose Injection). The 250-800-nm absorption spectra and the XRD patterns showed that both formulations contain a mineral core composed of iron oxyhydroxide in the beta-FeOOH mineral polymorph known as akaganeite. This was further confirmed for each formulation by imaging using TEM and AFM. The average core size for the nanoparticles, after dialysis to remove unbound or loosely bound carbohydrate, was approximately 3+/-2 nm for the iron-sucrose, and approximately 2+/-1 nm for the iron-gluconate. Each of the nanoparticles consists of a mineral core, surrounded by a layer of bound carbohydrate. The overall diameter of the average bead in the dialyzed preparations was approximately 7+/-4 nm for the iron-sucrose, and 3+/-1 nm for the iron-gluconate. Undialyzed preparations have particles with larger average sizes, depending on the extent of dilution of unbound and loosely bound carbohydrate. At a dilution corresponding to a final Fe concentration of 5 mg/mL, the average particle diameter in the iron-sucrose formulation was approximately 22+/-9 nm, whereas that of the iron-gluconate formulation was approximately 12+/ 5 nm. PMID- 15522404 TI - Coordination mode and oxidation susceptibility of nickel(II) complexes with 2' deoxyguanosine 5'-monophosphate and l-histidine. AB - The formation of binary and ternary complexes of Ni(II) with two biologically relevant molecules, 2'-deoxyguanosine 5'-monophosphate (dGMP) and l-histidine (histidine or His) was characterized by potentiometry and UV-visible spectroscopy. For dGMP, the mononuclear complexes with stoichiometries NiH(2)L(+), NiHL and NiL(-) were found. In the mixed system the ternary complexes NiH(2)LA, NiHLA(-) and NiLA(2-) were detected. In binary systems, the Ni(II) ion coordinates to dGMP through the N-7 atom of its purine ring and indirectly through a water molecule bonded to the phosphate group, while in ternary complexes Ni(II) is bonded to all three histidine donors and directly to the phosphate group of dGMP. Both binary and ternary complexes are susceptible to oxidation by H(2)O(2), with the increased formation of 8-oxo-dGMP in the ternary system. The toxicological relevance of these findings stems from possible disturbance by the major biological Ni(II)-His complex of the nucleotide pools homeostasis through the formation of ternary species and oxidation promotion, as well as from 8-oxo-dGMP capacity to inhibit enzymatic elimination of promutagenic oxidized nucleotides from such pools. PMID- 15522405 TI - Effect of a conjugated acridine moiety on the binding and reactivity of Cu(II)[9 acridinylmethyl-1,4,7-triazacyclononane] with DNA. AB - The DNA binding orientation and dynamic behavior of Cu(II) complexes of 1,4,7 triazacyclononane ([9]aneN(3)), 1, and an acridine conjugate, 2, were investigated by DNA fiber EPR (EPR=electron paramagnetic resonance) spectroscopy. Crystal and molecular structure of 2 were determined by X-ray diffraction. It has been shown that 1 binds to DNA in two different modes at room temperature; one species is rapidly rotating and the other is immobilized randomly on the DNA. The introduction of acridine to [9]aneN(3) fixed the [Cu([9]aneN(3))](2+) moiety of 2 in two different environments on the DNA: the g(mid R:mid R:) axis of one species (g( parallel)=2.26) is aligned perpendicularly to the DNA fiber axis whereas that of the other (g( parallel)=2.24) aligns<90 degrees with the DNA fiber axis. The different DNA binding structures of 1 and 2 are reflected also in their different efficiencies of DNA cleavage; 2 was found to be more effective both in oxidative and hydrolytic cleavage reactions. PMID- 15522406 TI - Enantiomeric and mesomeric mandelate complexes of molybdenum -- on their stereospecific formations and absolute configurations. AB - The stereospecific formation and absolute configuration of R-homocitrate coordinated FeMo-co in nitrogenase was mimicked through the structural analyses of a collection of enantiomeric and mesomeric mandelato molybdenum complexes, i.e., (NH(4))(2)[Mo(Delta)O(2)(R-mand)(2)]x3H(2)O (1a), (NH(4))(2)[Mo(Lambda)O(2)(S-mand)(2)]x3H(2)O (1b), (NH(4))(4)[Mo(Delta)O(2)(RS mand)(2)][Mo(Lambda)O(2)(RS-mand)(2)]x8H(2)O (2), (NH(4))(2)[W(Delta)O(2)(R mand)(2)]x2H(2)O (3a), (NH(4))(2)[W(Lambda)O(2)(S-mand)(2)]x2H(2)O (3b) (H(2)mand=mandelic acid, C(8)H(8)O(3)), which have been characterized by elemental analyses, optical rotation, circular dichroism, IR, NMR spectroscopes and X-ray single crystal studies. The R and S chiral mandelic acids induce the formations of the enantiomeric pair of chiral complexes, which are supported by the characterizations of optical rotation and circular dichroism. The configuration of the resulted metal center could be assigned as Delta or Lambda. While the RS racemic reagent yields only mesomeric compound. The Delta(R,R) complexes 1a and 3a are enantiomers of Lambda(S,S)-1b and 3b, respectively. Of the five complexes, Mo and W atoms are all hexa-coordinated by two cis-oxo groups and two bidentate mandelate ligands through the deprotonated alpha-alkoxyl and alpha-carboxyl groups, forming a stable five-membered chelated rings. The average Mo(VI)-O bond distances with alpha-alkoxyl and alpha-carboxyl are 1.944 and 2.210 A, respectively. Further comparison indicates that bonds of alpha-alkoxyl groups in the hydroxycarboxylato molybdenum complexes are much sensitive to the change in the oxidation state of molybdenum, which support the possible Mo activation model in FeMo-co through the protonation and cleavage of alpha-alkoxyl group in homocitrate ligand. PMID- 15522407 TI - Spectroscopic studies on the anticancer antibiotic Altromycin H and the interaction with copper(II) ions. AB - The antitumor antibiotic Altromycin H was studied using electronic absorption (UV Vis.) and circular dichroism (CD) spectroscopy. The dissociation constants of the phenolic groups on C(5) and C(11) were estimated as pK(1)=6.7 and pK(2)=11.8 at 25 degrees C, respectively, and a complete assignment of the CD and UV-Vis. bands is proposed. The interaction of Cu(II) ions with the Altromycin H has been also investigated by UV-Vis., CD and electron paramagnetic resonance (EPR) spectroscopy. A pH depended stepwise complex formation was observed. At pH<4 no copper-Altromycin H interactions were detected. At the 412, the predominant species, [Ru(II)(edta)NO](-), reacts according to [Ru(II)(edta)NO](-)+2OH(-)-->[Ru(II)(edta)NO(2)](3-), with K(eq)=1.0+/-0.4 x 10(3) M(-2) (I=1.0 M, NaCl; T=25.0+/-0.1 degrees C). The rate-law is first order in each of the reactants for most reaction conditions, with k(OH(-))=4.35+/-0.02 M(-1)s(-1) (25.0 degrees C), assignable mechanistically to the elementary step comprising the attack of one OH(-) on [Ru(II)(edta)NO](-), with subsequent fast deprotonation of the [Ru(II)(edta)NO(2)H](2-) intermediate. The activation parameters were DeltaH(#)=60+/-1 kJ/mol, DeltaS(#)=-31+/-3 J/Kmol, consistent with a nucleophilic addition process between likely charged ions. In the toxicity up-and-down tests performed with Swiss mice, no death was observed in all the doses administered (3-9.08 x 10(-5) mol/kg). The biodistribution tests performed with Wistar male rats showed metal in the liver, kidney, urine and plasma. Eight hours after the injection no metal was detected in the samples. The vasodilator effect of [Ru(II)(edta)NO](-) was studied in aortic rings without endothelium, and was compared with sodium nitroprusside (SNP). The times of maximal effects of [Ru(II)(edta)NO](-) and SNP were 2 h and 12 min, respectively, suggesting that [Ru(II)(edta)NO](-) releases NO slowly to the medium in comparison with SNP. PMID- 15522419 TI - Water-soluble platinum(II) complexes of diamine chelating ligands bearing amino acid type substituents: the effect of the linked amino acid and the diamine chelate ring size on antitumor activity, and interactions with 5'-GMP and DNA. AB - Six new Pt(II) complexes are described having the general formula PtCl(2)(LL), in which LL is a chelating diamine ligand bearing an amino acid as substituent. The amino acids chosen are l-alanine and its methyl ester, and l-phenylalanine. The compounds have been characterized using analytical and spectroscopic methods. The influence on the biological properties of the size of the chelate ring and the structure of the amino acid substituent has been studied. The effect of the presence of a carboxylic or carboxylate group on the amino acid C-terminus has also been determined. It is demonstrated by circular dichroism (CD) that the effect on the secondary structure of DNA induced by the six complexes differ from each other. In all cases, the interaction takes place at the N7 position of the purine bases, as shown by NMR monitoring. The general behavior of these platinum complexes, with one exception, is to uncoil the DNA from the B form to the C form. The interactions with 5'-GMP and DNA have been compared with their expected antitumour activity. The complexes with l-alanine and l-phenylalanine exhibit cytotoxic activity in HeLa and HL-60 cell lines, in a dose- and time-dependent manner. No cytotoxic activity of the methyl ester derivatives have been determined because of their low solubility in aqueous solution. PMID- 15522421 TI - Complexation of desferricoprogen with trivalent Fe, Al, Ga, In and divalent Fe, Ni, Cu, Zn metal ions: effects of the linking chain structure on the metal binding ability of hydroxamate based siderophores. AB - Complexes of the natural siderophore, desferricoprogen (DFC), with several trivalent and divalent metal ions in aqueous solution were studied by pH potentiometry, UV-Vis spectrophotometry and cyclic voltammetry. DFC was found to be an effective metal binding ligand, which, in addition to Fe(III), forms complexes of high stability with Ga(III), Al(III), In(III), Cu(II), Ni(II) and Zn(II). Fe(II), however, is oxidized by DFC under anaerobic conditions and Fe(III) complexes are formed. By comparing the results with those of desferrioxamine B (DFB), it can be concluded that the conjugated beta-double bond slightly increases the stability of the hydroxamate chelates, consequently increases the stability of mono-chelated complexes of DFC. Any steric effect by the connecting chains arises only in the bis- and tris-chelated complexes. With metal ions possessing a relatively big ionic radius (Cu(II), Ni(II), Zn(II), In(III)) DFC, containing a bit longer chains than DFB, forms slightly more stable complexes. With smaller metal ions the trend is the opposite. Also a notable difference is that stable trinuclear complex, [Cu(3)L(2)], is formed with DFC but not with DFB. Possible bio-relevance of the Fe(II)/Fe(III) results is also discussed in the paper. PMID- 15522420 TI - Cisplatin-mediated selective hydrolytic cleavage of methionine-containing peptides with neighboring serine or histidine residues. AB - The pH- and time-dependent reactions of the anticancer drug cisplatin, cis [PtCl(2)(NH(3))(2)], with the peptides Ac-Gly-Met-Gly-OH, Ac-Ser-Met-OH and Ac Met-His-OH (Gly=glycyl, Met=methionyl, Ser=seryl, His=histidyl) at 313 K have been investigated by high-performance liquid chromatography, nuclear magnetic resonance and mass spectrometry. In the major equimolar reaction pathway for Ac Gly-Met-Gly-OH, rapid anchoring at the methionine sulphur (kappaS) is followed by successive metalations of the methionine N(M) and glycyl N(G1) amide nitrogens in N-terminal direction to afford bidentate kappa(2)S,N(M) and tridentate kappa(3)S, N(M),N(G1) complexes. Cleavage of acetic acid at the second upstream amide bond is observed after 10 h leading to slow formation of [Pt(H-Gly-MetH(-1)-Gly-OH kappa(3)S,N(M),N(G1))(NH(3))](+) at pH<6. [Pt(H-Ser-MetH(-1)-OH kappa(3)S,N(M),N(S))(NH(3))](+) results from an analogous cisplatin-mediated regioselective hydrolytic cleavage reaction for Ac-Ser-Met-OH in moderately acid solution (pH<4). After passing through a minimum at pH 4.4, the concentration of the cleavage product in the reaction mixture after 500 h increases steadily on raising the pH and release of acetic acid is effectively quantitative for 7pH9.5. A competing mechanism involving nucleophilic attack of the serine side chain on the acetyl function can be inferred for pH>6 by the HPLC detection of a second intermediate kappa(3)S,N(M),N(S) species. In striking contrast, the reaction of cisplatin with Ac-Met-His-OH leads to release of acetylmethionine and formation of a final histidine product cis-[PtCl(H-His-OH-kappa(2)N3,N(H)) (NH(3))](+) at pH<6 by a kappaS-->kappa(2)S, N3-->kappa(3)S, N(H),N3-->kappa(2)N3,N(H)(H-His-OH) pathway. PMID- 15522422 TI - The first supramolecular orthovanadate receptor -- structural mimics of vanadium haloperoxidase. AB - Tris(2-guanidinium-ethyl)amine (1) was prepared as a supramolecular receptor of hydrogen orthovanadate (HVO(4)(2-)) to mimic the active site of vanadium haloperoxidase (V-HPO). Both (1)H and (51)V NMR titration indicated 1:1 complex (5) formation between (1) with HVO(4)(2-) with a binding constant of 1.1 x 10(3) M(-1). Similar as V-HPO, a UV band at 307 nm was observed upon binding of HVO(4)(2-) to (1). According to DFT calculations UV transitions >300 nm observed for both the enzyme and its mimic are due to V-N interactions. PMID- 15522424 TI - Phosphate ester hydrolysis is catalyzed by a bacterial transferrin: potential implications for in vivo iron transport mechanisms. AB - Two synergistic anions, p-nitrophenyl phosphate ester (NPP) and SO(4)(2-), were found to form new stable assemblies with Fe(3+) and a bacterial transferrin, FbpA (FbpA=ferric binding protein). Fe(3+)FbpA-SO(4) undergoes rapid anion exchange in the presence of NPP to form Fe(3+)FbpA-NPP. Formation of Fe(3+)FbpA-NPP was found to accelerate the rate of hydrolysis of the bound phosphate ester (k(hyd)=1.6 x 10(-6) s(-1) at 25 degrees C and pH 6.5) by >10(3) fold over the uncatalyzed reaction. These findings suggest a dual function for FbpA in vivo: transport of Fe(3+) across the periplasmic space to the inner membrane in certain gram negative bacteria and hydrolysis of periplasmic polyphosphates. PMID- 15522423 TI - Metal-induced folding of a designed metalloprotein. AB - The metal-induced assembly of a designed peptide-based rubredoxin model is described. The C16C19-GGY peptide has the sequence Ac K(IEALEGK)(2)(CEACEGK)(IEALEGK)GGY-amide in which the presence of the Cys-X-X-Cys metal binding domain of rubredoxin was used to place cysteine residues at the hydrophobic "a" and "d" positions upon formation of a homodimeric alpha-helical coiled-coil. Circular dichroism spectroscopy shows that the apopeptide exists as a random coil and assembles into a coiled-coil in the presence of Cd(2+). Metal binding is monitored by the appearance of a new LMCT band at 238 nm. UV-Vis titrations and SDS-PAGE experiments are used to show that this designed metalloprotein exists as a metal-bridged coiled-coil dimer. PMID- 15522425 TI - GMP and AMP as methyl radical traps in the reaction with pentaamminemethylcobalt(III). AB - C8 methylation of the title purine nucleotides was achieved in near neutral aqueous solution by reaction with (13)C enriched [Co(NH(3))(5)(CH(3))](2+). This cation decomposes upon dissolution in water by release of a methyl radical. The latter was identified by electron spin resonance (ESR) spectroscopy as the PBN/(13)C(.-)H(3) adduct (PBN=phenyl-N-tert-butylnitrone). When the purine nucleotide was used in large excess, the efficiency of the trapping by the C8 atom was determined by integration of the (13)C NMR signals to be 20-25% for GMP and 15-20% for AMP, respectively, at 37 degrees C. PMID- 15522426 TI - Inhibition of the Escherichia coli RecA protein: zinc(II), copper(II) and mercury(II) trap RecA as inactive aggregates. AB - In bacteria, the RecA protein plays important roles in a number of DNA recombination and repair processes, including homologous recombination, SOS induction and recombinational DNA repair. We have explored the idea that the Escherichia coli RecA protein's functions could be controlled by small molecules. We investigated the 2:1 complex of zinc(II) with 1,4-dithio-l-threitol (l-DTT) that inhibits the E. coli rho transcription terminator, which is a hexameric ATP motor protein and is structurally homologous to RecA. We found that both the complex and ZnCl(2) inhibit the single-stranded DNA-dependent ATPase activity of RecA at sub-millimolar concentrations. Investigation of a variety of metal dications (0.4 mM final concentration) determined that zinc(II), copper(II) and mercury(II) all induce the precipitation of RecA, while the dichloride salts of calcium, manganese, barium, cobalt, and nickel do not. The inhibition of RecA activity by Zn(II), Cu(II) and Hg(II) results from the metal-dependent initiation of RecA aggregation. These observations may have implications for the design of biophysical experiments requiring solid-phase RecA protein, for a more complete understanding of metal toxicities, and for the design of metal-chelate inhibitors of prokaryotic DNA repair. PMID- 15522428 TI - Characterization of responses to the antiandrogen flutamide in a short-term reproduction assay with the fathead minnow. AB - A short-term reproduction assay with the fathead minnow (Pimephales promelas) has been developed to detect chemicals with the potential to disrupt reproductive endocrine function controlled by estrogen- and androgen-mediated pathways. The objective of this study was to use the assay to characterize responses of fathead minnow reproductive endocrinology and physiology to the mammalian antiandrogen, flutamide. Male and female fish were exposed to nominal (target) concentrations of 50 and 500 microg flutamide/l for 21-days, following which plasma steroid and vitellogenin concentrations were determined and gonadal morphology assessed. Fecundity of the fish was significantly reduced by exposure to a measured test concentration of 651 microg flutamide/l. In addition, embryo hatch was significantly reduced at this concentration. Qualitative histological assessment of ovaries from females exposed to flutamide indicated a decrease in mature oocytes and an increase in atretic follicles. Testes of males exposed to flutamide exhibited spermatocyte degeneration and necrosis. Concentration dependent increases in plasma testosterone and vitellogenin concentrations were observed in the females. Flutamide also altered reproductive endocrinology of male fathead minnows. Males exposed to 651 microg flutamide/l exhibited elevated concentrations of beta-estradiol and vitellogenin. In summary, the results of this study with the fathead minnow demonstrate that flutamide affects reproductive endocrine function in fish and that the type of hormonal pattern and histopathology effects observed are consistent with an antiandrogenic mode-of action. Consequently, our findings suggest that the 21-day reproduction assay utilizing fathead minnows is a sensitive short-term screening method for the detection of endocrine-disrupting chemicals, including antiandrogens. PMID- 15522429 TI - cDNA-directed expression of a functional zebrafish CYP1A in yeast. AB - A cytochrome P450 1A (CYP1A) cDNA was isolated from an adult zebrafish (Danio rerio) library. The 2580-bp clone (GenBank Accession No. AF210727) contained a 62 bp 5'-unstranslated region (UTR), 1557-bp coding region and 962-bp 3'-UTR. The deduced 519-residue protein (calculated molecular weight 58,556, pI = 7.58) shared 74% identity with rainbow trout CYP1A and 57 and 54% identities with mouse and human CYP1A1s, respectively. The zebrafish CYP1A protein coding region was cloned into the pDONR201 entry vector and then transferred to a yeast expression vector pYES-DEST52. Expression of zebrafish CYP1A in Saccharomyces cerevisiae transformants was induced by galactose to a maximum level of 493 pmol CYP1A per mg microsomal protein or about 8 nmol/l of culture. Recombinant CYP1A protein expressed in yeast was mainly in the denatured P420 form under normal microsomal preparation conditions but when the oxygen concentration was reduced in the buffer by degassing and the yeast cells were maintained at less than 10 degrees C, the integrity of the CYP1A was preserved and it exhibited a characteristic reduced CO-difference spectrum maximum at 448 nm. The recombinant zebrafish CYP1A demonstrated 7-ethoxyresorufin O-deethylase (EROD) activity with an apparent Km (Km(app)) and Vmax values at 30 degrees C of 0.31 +/- 0.04 microM and 0.70 +/- 0.10 nmol/min/nmol CYP, respectively. The recombinant protein also metabolized benzo(a)pyrene with a Km(app) and Vmax values of 5.34 +/- 0.58 microM and 1.16 +/ 0.13 nmol/min/nmol CYP, respectively. These results show the recombinant expression of a functional zebrafish CYP in yeast and validated yeast as a host for heterologous expression of zebrafish CYP1A and potentially for other zebrafish CYPs. PMID- 15522430 TI - Effects of endocrine-disrupting chemicals on expression of ubiquitin C-terminal hydrolase mRNA in testis and brain of the Japanese common goby. AB - We investigated the effects of endocrine-disrupting chemicals (EDCs) on the expression of ubiquitin C-terminal hydrolase (UCH) mRNA in the testis and brain of the Japanese common goby, Acanthogobius flavimanus. The cDNA sequence of goby UCH contained an open reading frame encoding 220 amino acid residues (M(r)=24,223) with 51.3% overall sequence identity with human and mouse UCHL1. A competitive PCR assay was used to quantify the levels of UCH mRNA in the testis and brain of male gobies after exposure to bisphenol A, nonylphenol, or estradiol 17beta for 3 weeks. Exposure to estradiol-17beta at a nominal concentration of 100 ng/L induced significant increase in UCH mRNA levels in both testis and brain (P<0.05), whereas exposure to nonylphenol induced a significant decrease in UCH mRNA levels in the testis (P<0.01). These results suggest that EDCs can either positively or negatively regulate UCH mRNA levels. PMID- 15522431 TI - Bioavailability of silver and its relationship to ionoregulation and silver speciation across a range of salinities in the gulf toadfish (Opsanus beta). AB - Silver is taken up as a Na(+) analog (Ag(+)) by freshwater organisms, but little is known about its bioavailability in relation to salinity. Adult Opsanus beta were acclimated to 2.5, 5, 10, 20, 40, 60, 80, and 100% seawater (Cl(-)=545 mM) and exposed for 24 h to 2.18 microg L(-1) silver as (110m)Ag-labelled AgNO(3), a concentration close to the U.S. EPA marine criterion and less than 0.1% of the acute 96-h LC50 in seawater. Plasma osmolality, Na(+), and Cl(-) remained approximately constant from 100% down to 20-40% seawater, thereafter declining to 89% (osmolality) and 82% (Na(+), Cl(-)) of seawater values at the lowest salinity (2.5% seawater), while plasma Mg(2+) was invariant. Ionic measurements in intestinal fluids and urine supported the view that above the isosmotic point (about 32% seawater), toadfish drink the medium, absorb Na(+), Cl(-), and water across the gastrointestinal tract, actively excrete Na(+) and Cl(-) across the gills, and secrete Mg(2+) into the urine. Below this point, toadfish appear to stop drinking, actively take up Na(+) and Cl(-) at the gills, and retain ions at the kidney. Silver accumulation varied greatly with salinity, by nine-fold (whole body), 26-fold (gill tissue), and 18-fold (liver), with the maxima occurring in 2.5% seawater, the minima in 40% seawater (close to the isosmotic point), and slightly greater values at higher salinities. Highest silver concentrations occurred in liver, second highest in gills, intermediate concentrations in kidney, spleen, and gastrointestinal tissues, and lowest in swim bladder and white muscle, though patterns changed with salinity. There were substantial biliary but minimal urinary levels of silver. The salinity-dependent pattern of silver accumulation best correlated with the abundance of the neutral complex AgCl(0), though the presence of small amounts of Ag(+) at the lowest salinities may also have been important. In contrast, silver accumulation in the esophagus stomach was greatest in 100% seawater and least at the isosmotic salinity (five fold variation), a pattern probably explained by drinking and silver uptake into the blood through the gills. Models of silver bioavailability across salinity must consider the presence of silver-binding ligands on both gills and gastrointestinal tract, changing silver speciation, and the changing ionoregulatory physiology of the organism. PMID- 15522432 TI - Acute toxicity of hexavalent chromium in isolated teleost hepatocytes. AB - Acute toxic effects of hexavalent chromium [Cr(VI)], a widely recognised carcinogenic, mutagenic and redox active metal, were investigated in isolated hepatocytes of goldfish (Carassius auratus). Exposure to 250 microM Cr(VI) induced a significant decrease of cell viability from 94% in controls to 88% and 84% after 30 min and 4 h of exposure, respectively. Cr-toxicity was associated with a concentration-dependent stimulation of the formation of reactive oxygen species (ROS). As one potential source of ROS formation we identified the lysosomal Fe(2+) pool, since the ferric ion chelator deferoxamin inhibited ROS formation by approximately 15%. Lysosomal membranes remained nevertheless intact during Cr-exposure, as determined from neutral red retention in this compartment. Another significant source of ROS appear to be the mitochondria, where a presumably uncoupled increase of respiration by 20-30% was triggered by the metal. Inhibition of mitochondrial respiration by cyanide caused an approximately 40% decrease of Cr-induced ROS-formation, whereas the uncoupling agent carbonyl cyanide m-chlorophenyl hydrazine was without effect. Cellular Ca(2+) homeostasis was not disturbed by Cr(VI) and thus played no role in this scenario. Overall, our data show that Cr(VI) is acutely toxic to goldfish hepatocytes, and its toxicity is associated with the induction of radical stress, presumably involving lysosomes and mitochondria as important sources of ROS formation. PMID- 15522433 TI - Cassette-based presentation of SIV epitopes with recombinant gas vesicles from halophilic archaea. AB - In earlier studies we demonstrated recombinant gas vesicles from Halobacterium sp. NRC-1, expressing a model six amino acid insert, or native vesicles displaying chemically coupled TNP, each were immunogenic, and antigenic. Long lived responses displaying immunologic memory were elicited without exogenous adjuvant. Here we report the generation and expression of cassettes containing SIV derived DNA. The results indicate a cassette-based display/delivery system derived from recombinant halobacterial gas vesicle genes is highly feasible. Data specifically support four conclusions: (i) Recombinants carrying up to 705 bp of SIV DNA inserted into the gvpC gene form functional gas vesicles; (ii) SIV peptides contained as part of the expressed recombinant, surface exposed GvpC protein are recognized by antibody elicited in monkeys exposed to native SIV in vivo; (iii) in the absence of adjuvant, mice immunized with the recombinant gas vesicle (r-GV) preparations mount a solid, titratable antibody response to the test SIV insert that is long lived and exhibits immunologic memory; (iv) recombinant organelles, created through the generation of cassettes encoding epitopes inserted into the gvpC DNA, can be used to construct a multiepitope display (MED) library, a potentially cost effective vehicle to express and deliver peptides of SIV, HIV or other pathogens. PMID- 15522434 TI - Impact of intrinsic DNA structure on processing of plasmids for gene therapy and DNA vaccines. AB - Several non-Watson Crick DNA structures have been discovered to date, which may be incorporated into future plasmid constructs for gene therapy and DNA vaccine products. In this study, intrinsic DNA structures were included at a defined point in a 2.9 kb plasmid, and their effects on cell growth rate, total plasmid yield, and topology (i.e. the relative proportions of supercoiled plasmid, open circular and linear forms), were determined. The stability of the inserted sequences were assessed using gel electrophoresis. Z-DNA was shown to be unstable in a batch Escherichia coli DH1 production system grown in complex medium. Encouragingly other sequences studied (triplex, bend and quadruplex) did not cause spontaneous deletions, and no detrimental effect was found on growth rate or on total plasmid yield; indicating that such sequences could be included in future DNA products without any detrimental effect on plasmid yields; although the intra molecular triplex studied significantly decreased the proportion of supercoiled species. PMID- 15522435 TI - Design of immobilised dextransucrase for fluidised bed application. AB - Immobilisation of dextransucrase from Leuconostoc mesenteroides NRRL B-512F in alginate is optimised for applications in a fluidised bed reactor with high concentrated sugar solutions, in order to allow a continuous formation of defined oligosaccharides as prebiotic isomalto-oligosaccharides. Efficient design of fluidised bed immobilised biocatalyst in high density solutions requires particles with elevated density, high effectiveness and both thermal and mechanical stability. Inert silica flour/sand (Mikrosil 300) as supplement turned out to be best suited for increasing the density up to 1400 kg m(-3) of the alginate beads and generating a stable expanded bed without diffusional restrictions. Kinetic investigations demonstrate that low effectiveness of immobilised enzyme due to close association to dextranpolymers (dextran content of enzyme preparation >90%) is compensated by reducing the particle size and/or by decreasing the dextran content. A low dextran content (5%) is sufficient to immobilise and stabilise the enzyme, thus diffusional limitation is reduced essentially while operational stability is maintained. Fluidisation behaviour and bed expansion proved to be appropriate for the intended application. Both calculated and measured expansion coefficients showed good agreement for different conditions. PMID- 15522436 TI - Sorbitol prevents the self-aggregation of unfolded lysozyme leading to and up to 13 degrees C stabilisation of the folded form. AB - We present a calorimetric investigation of stabilisation of hen egg-white lysozyme with sorbitol in the pH range 3.8-10.5. Differential scanning calorimetry and steady-state fluorescence were used to determine the denaturation temperatures of lysozyme as a function of sorbitol concentration. The fluorescence data were collected in the presence of 2M urea to lower the melting point of the protein to an observable range of the instrument. The effect of sorbitol on the activation energy of unfolding was investigated by scanrate studies. The effect of sorbitol lysozyme interaction was investigated using isothermal titration calorimetry. The titration experiments were performed with folded as well as unfolded lysozyme to investigate in more detail the nature of the interaction. The data obtained in those experiments show a remarkable stabilisation effect of sorbitol. We observed a 4.0 degrees C increase in the Tm for 1 M sorbitol in the pH range 3.8-8.5 by scanning calorimetry. The effect increases dramatically at pH 9.5 where we observe a 9.5 degrees C stabilisation. An increase in the sorbitol concentration to 2 M stabilises lysozyme by 11.3-13.4 degrees C in the pH range 9.5-10.5. In the absence of urea, no significant effects of sorbitol were observed on the activation energy for unfolding for lysozyme at pH 4.5. This indicates together with the results from the titration experiments that sorbitol may stabilise the folded form of lysozyme by destabilising the unfolded form of lysozyme. At pH values at and above lysozyme's pI (approximately 9.3), the unfolding of the protein is accompanied with a substantial amount of self-aggregation seen in the calorimetry experiments in the ratio of DeltaH(cal)/DeltaH(vH). In the presence of sorbitol, the self aggregation was counterbalanced by higher sorbitol concentrations. These results strongly suggest a negative influence of sorbitol on the unfolded form of lysozyme and thereby stabilising the native form. PMID- 15522437 TI - Aspergillus fumigatus CY018, an endophytic fungus in Cynodon dactylon as a versatile producer of new and bioactive metabolites. AB - Aspergillus fumigatus CY018 was recognized as an endophytic fungus for the first time in the leaf of Cynodon dactylon. By bioassay-guided fractionation, the EtOAc extract of a solid-matrix steady culture of this fungus afforded two new metabolites, named asperfumoid (1) and asperfumin (2), together with six known bioactive compounds including monomethylsulochrin, fumigaclavine C, fumitremorgin C, physcion, helvolic acid and 5alpha,8alpha-epidioxy-ergosta-6,22-diene-3beta-ol as well as other four known compounds ergosta-4,22-diene-3beta-ol, ergosterol, cyclo(Ala-Leu) and cyclo(Ala-Ile). Through detailed spectroscopic analyses including HRESI-MS, homo- and hetero-nuclear correlation NMR experiments (HMQC, COSY, NOESY and HMBC), the structures of asperfumoid and asperfumin were established to be spiro-(3-hydroxyl-2,6-dimethoxyl-2,5-diene-4-cyclohexone-(1,3') 5'-methoxyl-7'-methyl-(1'H, 2'H, 4'H)-quinoline-2',4'-dione) and 5-hydroxyl-2-(6 hydroxyl-2-methoxyl-4-methylbenzoyl)-3,6-dimethoxyl-benzoic methyl ester, respectively. All of the 12 isolates were subjected to in vitro bioactive assays against three human pathogenic fungi Candida albicans, Tricophyton rubrum and Aspergillus niger. As a result, asperfumoid, fumigaclavine C, fumitremorgin C, physcion and helvolic acid were shown to inhibit C. albicans with MICs of 75.0, 31.5, 62.5, 125.0 and 31.5 microg/mL, respectively. PMID- 15522438 TI - Use of volatile fatty acids from an acid-phase digester for denitrification. AB - This study investigates the ability of naturally-produced volatile fatty acids (VFAs) to act as a carbon source for the removal of nitrate-Nitrogen concentrations in the range of 20 to 200 mg/L. The VFAs were generated from an anaerobic digester treating a 1:1 mixture of starch-rich industrial and municipal wastewater. The experiments were carried out at an ambient temperature of 22+/-2 degrees C using 600 mL batch reactors containing VFA-rich effluent from the digester and prepared primary sludge. Nitrates added in the form of a 1 M potassium nitrate solution were observed to completely disappear in conjunction with significant recovery of pH and alkalinity (i.e. biological denitrification). In all cases, denitrification followed zero-order kinetics with an average rate constant of 2.67 mg/[Lh] and a mean specific denitrification rate of 0.0111 g NOx N/g volatile suspended solids (VSS) per day. The denitrifier population had a preference for acetic acid which occurred in the largest quantity and was exhausted the most rapidly. The next two most abundant VFAs were butyric acid and propionic acid, which were consumed only after acetate concentrations began to decline. The third "choice" of the denitrifiers was valeric acid, thus it appears that denitrifying bacteria have a sequential preference for VFAs. In addition, they prefer VFAs over other more complex soluble organic carbon forms. PMID- 15522439 TI - Enhancement of bioactivity of Saccharomyces cerevisiae alpha-mating factor by attachment of sugar moiety to glutamine residue. AB - We prepared yeast Saccharomyces cerevisiae alpha-mating factor, a 13-amino acid pheromone produced by haploid alpha-cells, bound with glucose or N acetylglucosamine at the fifth glutamine residue from the N-terminal by the chemical method of peptide synthesis. It was found that the bioactivity of glucosyl alpha-mating factor was higher than that of native alpha-mating factor. However, it was slightly lower than that of N-acetylglucosaminyl alpha-mating factor. This suggested that the N-acetylamino residue might play some important role in the enhancement of the bioactivity of alpha-mating factor. However, CD spectra analysis of alpha-mating factor and its derivatives demonstrated that their structures were almost identical. On the other hand, we attached a sialo complex type oligosaccharide to N-acetylglucosamine or its glucose residue by means of the transglycosylation activity of endo-beta-N-acetylglucosaminidase from Mucor hiemalis (Endo-M). The attachment of the oligosaccharide to both alpha mating factors reduced their activities. However, enzymatical trimming of the sialo complex type oligosaccharide recovered its activity. PMID- 15522440 TI - The effect of lactate addition on the growth of Penicillium camembertii on glutamate. AB - The effect of an additional carbon source, lactate, on Penicillium camembertii growth on glutamate as both carbon and nitrogen sources was examined. Glutamate (and lactate) was present in excess in both media. Throughout the whole culture, similar growth time-courses were recorded on both media, indicating the absence of a lactate effect on growth. During the first part of growth, corresponding to an increasing amount of viable biomass, the rate of glutamate consumption remained high, as well as the related ammonium production, indicating its use as a carbon source in addition to being nitrogen source. The low growth rates recorded during the last part of growth resulted in low glutamate consumption, while lactate consumption continued mainly by a maintenance mechanism for the energy supply. A clear differentiation appeared therefore between the carbon source and the energy source: glutamate was mainly used as C source (and N source) for biosynthesis, while lactate was mainly assimilated for energy supply. Carbon and nitrogen yield examinations confirmed this result. Indeed, the C/N ratio found for P. camembertii cellular material (8.14) was about twice that of glutamate (4.29). From this, about half of the available nitrogen was used for biomass formation during growth on glutamate-lactate based medium, as experimentally confirmed (constant yield nitrogen from biomass on nitrogen from glutamate was found (0.49), while the excess nitrogen was released as ammonium). The constant and close to unit (0.99) yield carbon from CO2 on carbon from lactate, also recorded during growth on glutamate-lactate based medium, confirmed that lactate was mainly used as an energy source. PMID- 15522441 TI - CellMAC: a novel technology for encapsulation of mammalian cells in cellulose sulfate/pDADMAC capsules assembled on a transient alginate/Ca2+ scaffold. AB - Microencapsulation of desired mammalian cell phenotypes in biocompatible polymer matrices represents a powerful technology for cell-based therapies and biopharmaceutical manufacturing of protein therapeutics. We have pioneered a novel jet break-up-compatible process for encapsulation of mammalian cells in cellulose sulfate (CS)/poly-diallyl-dimethyl-ammoniumchloride (pDADMAC) (CellMAC) capsules. CS and pDADMAC polymerize on a transient ad hoc co-assembled Ca2+/alginate scaffold and form homogenous capsules following dissolution of the alginate core by Ca2+ chelating agents. CellMAC capsules exhibited excellent mechanical properties and showed a molecular weight cut-off between 43 and 77kDa. Chinese hamster ovary cells engineered for constitutive production of the glycohormone erythropoietin reached high viable cell densities when grown inside CellMAC capsules, while specific erythropoietin (EPO) productivities matched those of conventional non-encapsulated control cultures. CellMAC-encapsulated EPO production cell lines induced increased EPO serum levels when implanted intraperitoneally into mice and provided robust glycoprotein production during standard stirred-tank bioreactor operation. We expect the CellMAC technology to foster advances in therapeutic encapsulation of engineered cell lines as well as manufacturing of protein pharmaceuticals. PMID- 15522442 TI - Respiratory syncytial virus and other pneumoviruses: a review of the international symposium--RSV 2003. AB - The Respiratory Syncytial Virus 2003 symposium took place from 8th-11th November 2003 in Stone Mountain, Georgia, and brought together more than 200 international investigators engaged in RSV research. RSV biology, pathogenesis, and clinical data, as well as RSV vaccines and antivirals, were addressed in the meeting, and this review will aim to briefly summarize and discuss the implications of new findings. The meeting also served as the inauguration of the Robert M. Chanock Award for lifetime achievement in RSV research, an award named in honor of the person who started the field of RSV research by recovering the first human RS virus from infants with severe bronchiolitis in 1956. PMID- 15522443 TI - Cloning and sequence analysis of dsRNA segments 5, 6 and 7 of a novel non-group A, B, C adult rotavirus that caused an outbreak of gastroenteritis in China. AB - A diarrhoeal outbreak among adults in China was caused by a new rotavirus, termed ADRV-N, that does not react with antisera directed against group A, B or C rotaviruses [Zhonghua Liu Xing Bing Xue Za Zhi (Chin. Epidemiol.) 19 (1998) 336]. ADRV-N can be propagated in cell cultures [Zhonghua Yi Xue Za Zhi (Natl. Med. J. China) 82 (2002) 14]. We present the complete sequences for ADRV-N genome segments 5 and 6, and a full ORF sequence of genome segment 7. The deduced amino acid sequences suggest that these segments encode NSP1, VP6 and NSP3, respectively. These three ADRV-N genome segments have a unique -ACCCC-3' terminal sequence. The 5'-GG- terminus of segments 5 and 6 is the same as that of other rotaviruses. The amino acid similarity between VP6 and NSP3 of ADRV-N and the cognate sequences of their closest counterpart, group B IDIR, was 37 and 35%, respectively. The ADRV-N NSP1 has a double-stranded RNA binding motif (DSRM) and a putative autoproteolytic cleavage motif upstream from the DSRM. The putative ADRV-N NSP3 has a truncated C-terminus compared to the cognate protein of group B rotaviruses. All the available data demonstrate that ADRV-N differs significantly from the known rotaviruses and strongly suggest that ADRV-N is the first recognized member of a new group of rotaviruses infecting humans. PMID- 15522444 TI - Changes in the human immunodeficiency virus V3 region that correspond with disease progression: a meta-analysis. AB - In order to determine the changes in the human immunodeficiency virus type-1 (HIV 1) envelope that corresponds with disease progression, a meta-analysis of viral forms was performed using HIV-1 sequences obtained from GenBank. Studies were selected that included longitudinally derived V3 envelope region sequences from multiple time points along with CD4 values as a marker of disease progression. Studies with a total of 58 subjects, 327 time points, and 380,000 total amino acid residues were included in this meta-analysis. Changes at specific amino acid sites over the course of disease progression stages were analyzed. The most common specific changes were found at amino acid sites 324D to N, 306S/G to R, and 360N to R. Other sites had changes from one amino acid type to another including the appearance of a basic form at 327, a charged form at 319, and 320D/E changing to basic or neutral. The timing of these changes was contrasted to CD4 decline with changes at 324 and 327 appearing before and 306, 320, and 319 appearing after the initiation of CD4 decline. PMID- 15522445 TI - Lactate dehydrogenase-elevating virus induces apoptosis in cultured macrophages and in spinal cords of C58 mice coincident with onset of murine amyotrophic lateral sclerosis. AB - Age-dependent poliomyelitis (ADPM) or murine amyotrophic lateral sclerosis (ALS) is a murine paralytic disease triggered in immunosuppressed genetically susceptible mice by infection with the arterivirus lactate dehydrogenase elevating virus (LDV). This disease provides an animal model for ALS, affecting anterior horn neurons and resulting in neuroparalysis 2-3 weeks after LDV infection. We have tested the hypothesis that spinal cord apoptosis is a feature of the LDV-induced murine ALS, since apoptosis is postulated to be a causal factor in human ALS. Gene microarray analyses of spinal cords from paralyzed animals revealed upregulation of several genes associated with apoptosis. Spinal cord apoptosis was investigated further by TUNEL and activated caspase-3 assays, and was observed to emerge concurrent with paralytic symptoms in both neuronal and non-neuronal cells. Caspase-3-dependent apoptosis was also triggered in cultured macrophages by neurovirulent LDV infection. Thus, virus-induced spinal cord apoptosis is a pre-mortem feature of ADPM, which affects both neuronal and support cells, and may contribute to the pathogenesis of this ALS-like disease. PMID- 15522446 TI - Rescue of Ebola virus from cDNA using heterologous support proteins. AB - Using the infectious clone for Zaire ebolavirus, the functional specificity of viral proteins of the ribonucleoprotein complex in transcription/replication was investigated by substituting them with heterologous proteins derived from closely (Reston ebolavirus) and distantly related filoviruses (Marburgvirus). The data clearly demonstrated that transcription/replication are neither strictly species specific nor genus-specific. Protein interactions between the nucleoprotein NP and the virion protein VP35 and the polymerase L and VP35 seemed to be the most critical steps. In contrast to previous data, viral proteins were able to target heterologous filovirus RNA. Together these results indicated that protein-protein interactions are more critical than protein-RNA interactions. PMID- 15522447 TI - Infectious salmon anemia virus (ISAV) genomic segment 3 encodes the viral nucleoprotein (NP), an RNA-binding protein with two monopartite nuclear localization signals (NLS). AB - Infectious salmon anemia virus (ISAV) is the type species of the genus Isavirus belonging to the Orthomyxoviridae, and causes serious disease in Atlantic salmon (Salmo salar). This study presents the expression and functional analysis of the ISAV genome segment 3, and provides further evidence that it encodes the viral nucleoprotein (NP). The encoded protein was expressed in a baculovirus system, and Western blot analysis showed that it corresponds to the 66-71 kDa structural protein previously found in purified ISAV preparations. RNA-binding activity was established by the interaction of viral and recombinant NP with single-stranded RNA transcribed in vitro. Immunofluorescence studies of infected cells showed the ISAV NP to be an early protein. It locates to the nucleus of infected cells before it is transported to the cytoplasm prior to virus assembly. A similar localization pattern was observed in cells transfected with the NP gene, confirming that the encoded protein has an intrinsic ability to be imported into the nucleus. Two monopartite nuclear localization signals (NLS) at amino acids (230)RPKR(233) and (473)KPKK(476) were identified by computer analysis, and validated by site-directed mutagenesis. In contrast to other orthomyxovirus-NPs, that have several NLSs that function independent of each other, both NLSs had to be present for the ISAV NP protein to be transported into the nucleus, indicating that these motifs cooperate to target the protein to the nucleus. PMID- 15522448 TI - Complete sequences of 3' end coding region for structural protein genes of turkey coronavirus. AB - Overlapping fragments of genomic RNA spanning 6963 nucleotides from 5' end of spike (S) protein gene to 3' end of nucleocapsid (N) protein gene of turkey coronavirus (TCoV) were amplified by reverse-transcription-polymerase chain reaction (RT-PCR). The primers were derived from the corresponding sequences of infectious bronchitis virus (IBV). The PCR products were cloned and sequenced and their nucleic acid structure and similarity to published sequences of other coronaviruses were analyzed. Sequencing and subsequent analysis revealed 9 open reading frames (ORFs) representing the entire S protein gene, tricistronic gene 3, membrane (M) protein gene, bicistronic gene 5, and N protein gene in the order of 5'-3'. The overall nucleic acid structures of these encoding regions of TCoV were very similar to the homologous regions of IBV. The consensus transcription regulating sequence (TRS) of IBV, CT(T/G)AACAA, was highly conserved in TCoV genome at the levels of nucleotide sequence and location in regarding to the initiation codon of individual genes. Pair-wise comparison of gene 3, M gene, gene 5, or N gene sequences with their counterparts of IBV revealed high levels (82.1-92.0%) of similarity. Phylogenetic analysis based on the deduced amino acid sequences of S, M, or N protein demonstrated that TCoV was clustered within the same genomic lineage as the IBV strains while all the other mammalian coronaviruses were grouped into separate clusters corresponding to antigenic groups I or II. There were substantial differences of S protein sequence between TCoV and IBV with only 33.8-33.9% of similarity. PMID- 15522450 TI - Selection and characterization of peptides specifically binding to HIV-1 psi (psi) RNA. AB - The packaging of HIV genomic RNA is mediated by a specific interaction between a nucleocapsid (NC) protein and packaging signal (psi, psi) RNA sequence. However, this interaction can be inhibited by the presence of peptides or proteins that specifically bind to the psi sequence. The 125-base-long psi RNA comprises a specific secondary structure that can be recognized by certain peptide sequences. Accordingly, the current study presents a method for selecting such peptides from a phage-displayed peptide library and characterization of resulting peptides in vitro. The RNA was covalently immobilized in a Covalink module using a carbodiimide condensation reaction at its 5'-end, leaving the proper secondary structure exposed and readily accessible. A phage display random peptide library was then screened against the RNA structure, and after five rounds of biopanning, enriched peptide sequences and conserved amino acid frames appeared. One of the enriched peptides was tested and shown to bind to psi RNA in a dose-dependent manner, plus it competed effectively with the NC protein as regards binding with the target RNA. PMID- 15522449 TI - Nipah virus glycoprotein: production in baculovirus and application in diagnosis. AB - A method for serological diagnosis of Nipah virus (NiV) is described. DNA encoding truncated G protein of NiV was cloned into the pFastBac HT vector, and the fusion protein to His-tag was expressed in insect cells by recombinant baculovirus. The resulting His-G recombinant fusion protein was purified by affinity chromatography and used as the coating antigen for serological testing by indirect enzyme-linked immunosorbant assay (ELISA). When tested against a panel of swine serum samples, the recombinant G protein-based ELISA successfully discriminated all 40 samples previously determined to be serum neutralizing test (SNT) positive from 11 SNT negatives samples. The data show that the recombinant G protein exhibits the antigenic epitopes and conformation necessary for specific antigen-antibody recognition. The main advantage of the recombinant G protein based NiV ELISA compared to an ELISA using whole virus antigen is the use of a single antigenic protein instead of inactivated whole virus which is required to be prepared under high risk and cost. This test is suitable for routine diagnosis of NiV and also for epidemiological surveys as it allows highly reliable testing of a large number of sera rapidly. PMID- 15522451 TI - Towards the reconstruction of the bilaterian ancestral pre-MHC region. AB - In this article, we describe how we reconstructed a precise, minimal proto-MHC region in the ancestor of euchordates, which was based on a comparison of the MHC paralogy group of vertebrate with the MHC-like chromosome of cephalochordates. This deduced ancestral region was compared with the genomes of extant species, other deuterostomes and protostomes. Our analysis revealed statistically significant traces of conservation in these species, suggesting that a proto-MHC region existed at the origin of all bilaterian species. We also propose a new approach to reconstruct ancestral genomes, which combines both stringent statistical testing and phylogenetic analysis. PMID- 15522452 TI - Assigning pathogenicity to mitochondrial tRNA mutations: when "definitely maybe" is not good enough. AB - Some mutations in mitochondrial tRNA (mt-tRNA) genes cause devastating disease, whereas others have no clinical consequences. We understand little of the factors determining the pathogenicity of specific mt-tRNA mutations, making prediction of clinical outcome extremely difficult. Using extensive sequence databases, we compared the characteristics of neutral variations with those of pathogenic mutations. We recommend that the location of the proposed mutation within the secondary structure of the mt-tRNA molecule and the disruption it causes to Watson-Crick base pairing should be considered when assessing the pathological significance of a novel mt-tRNA mutation. PMID- 15522453 TI - The comparative method for evaluating fossil calibration dates: a reply to Hedges and Kumar. PMID- 15522454 TI - Wide genome comparisons reveal the origins of the human X chromosome. AB - The eutherian X chromosome has one of the most conserved gene arrangements in mammals. Although earlier comparisons with distantly related mammalian groups pointed towards separate origins for the short and long arms, much deeper comparisons are now possible using draft sequences of the chicken genome, in combination with genome sequences from pufferfish and zebrafish. This enables surprising new insights into the origins of the mammalian X chromosome. PMID- 15522455 TI - Regulation of RNA function by aminoacylation and editing? AB - The chemical modification of nucleic acids is a ubiquitous phenomenon. Aminoacylation of tRNAs by aminoacyl-tRNA synthetases (ARSs) is a reaction essentially devoted to protein synthesis but it is used also as an emergency mechanism to recycle stalled ribosomes, and it is required for genome replication in some RNA viruses. In several aminoacyl-tRNA synthetases a correction mechanism known as editing is present to prevent aminoacylation errors. Genome data reveal a growing number of open reading frames encoding ARS-like proteins. This strongly suggests the existence of a widespread and nonconventional machinery for aminoacylation and editing. Here we review the different biological functions of aminoacylation and editing; also we propose an evolutionary scenario for the origin of these two reactions, and hypothesize an extant role for RNA charging and editing outside the genetic code. PMID- 15522456 TI - Evolutionary dynamics of transposable elements at the centromere. AB - Transposable elements are the single most abundant class of genetic material in higher eukaryotes. These elements show a genome-wide distribution but are found in disproportionate abundance at the centromeric and/or pericentric regions in a wide range of phylogenetic species. We propose at least three possible ways in which these elements could have directly contributed to the evolution of the architecture and function of the centromere in various organisms. An "extradition" mechanism also appears to have evolved, which enables the developing or established centromere to deal with the potentially disruptive effects of any subsequently arising transposable elements. PMID- 15522457 TI - MicroRNAs and the regulation of cell death. AB - Programmed cell death, or apoptosis, is ubiquitous, both during development and in the adult. Many components of the evolutionarily conserved machinery that brings about and regulates cell death have been identified, and all of these are proteins. However, in the past three years it has become clear that roughly 1% of predicted genes in animals encode small noncoding RNAs known as microRNAs, which regulate gene function. Here we review the recent identification of microRNA cell death regulators in Drosophila, hints that such regulators are also likely to exist in mammals, and more generally the approaches and tools that are now available to probe roles for noncoding RNAs in the control of cell death. PMID- 15522458 TI - An expanding genetic code. AB - More than 30 novel amino acids have been genetically encoded in response to unique triplet and quadruplet codons including fluorescent, photoreactive and redox active amino acids, glycosylated and heavy atom derived amino acids in addition to those with keto, azido and acetylenic chains. In this article, we describe recent advances that make it possible to add new building blocks systematically to the genetic codes of bacteria, yeast and mammalian cells. Taken together these tools will enable the detailed investigation of protein structure and function, which is not possible with conventional mutagenesis. Moreover, by lifting the constraints of the existing 20-amino-acid code, it should be possible to generate proteins and perhaps entire organisms with new or enhanced properties. PMID- 15522459 TI - Mammalian phylogenomics comes of age. AB - The relatively new field of phylogenomics is beginning to reveal the potential of genomic data for evolutionary studies. As the cost of whole genome sequencing falls, anticipation of complete genome sequences from divergent species, reflecting the major lineages of modern mammals, is no longer a distant dream. In this article, we describe how comparative genomic data from mammals is progressing to resolve long-standing phylogenetic controversies, to refine dogma on how chromosomes evolve and to guide annotation of human and other vertebrate genomes. PMID- 15522460 TI - Genetics, statistics and human disease: analytical retooling for complexity. AB - Molecular biologists and geneticists alike now acknowledge that most common human diseases with a genetic component are likely to have complex etiologies. Yet despite this belief, many statistical geneticists continue applying, in slightly new and different ways, methodologies that were developed to dissect much simpler etiologies. In this article, we characterize, with examples, the various factors that can complicate genetic analysis and demonstrate their shared features and how they affect genetic analysis. We describe a variety of approaches that are currently available, revealing methodological gaps and suggesting new directions for method development. Finally, we propose a comprehensive two-step approach to analysis that systemically addresses the different genetic factors that are likely to underlie complex diseases. PMID- 15522461 TI - One neuron-one receptor rule in the mouse olfactory system. AB - Similar to the expression of antigen receptor genes in lymphocytes, the mammalian odorant receptor (OR) genes are expressed in a mutually exclusive and monoallelic manner in olfactory sensory neurons (OSNs). DNA rearrangement has long been regarded as a possible mechanism for the allelic exclusion of the OR genes. However, mice cloned from mature OSN nuclei expressed the full repertoire of ORs, and the possibility of irreversible gene translocation was excluded as a mechanism to activate a single OR gene in each OSN. How is allelic exclusion achieved in the olfactory system? Recent transgenic experiments indicated an inhibitory role of the OR protein in preventing further activation of other OR genes. Stochastic activation of an OR gene and negative-feedback regulation by the OR gene product might ensure the maintenance of the one neuron-one receptor rule in the mammalian olfactory system. PMID- 15522462 TI - The prevalence of abnormal glucose regulation in patients with coronary artery disease across Europe. PMID- 15522463 TI - Decline in ventricular function and clinical condition after Mustard repair. PMID- 15522464 TI - Tissue Doppler imaging for the pre-clinical diagnosis of cardiomyopathy. PMID- 15522465 TI - Coronary flow: a new asset for the echo lab? PMID- 15522466 TI - The prevalence of abnormal glucose regulation in patients with coronary artery disease across Europe. The Euro Heart Survey on diabetes and the heart. AB - AIM: The objective behind the Euro Heart Survey on diabetes and the heart was to study the prevalence of abnormal glucose regulation in adult patients with coronary artery disease (CAD). METHODS AND RESULTS: The survey engaged 110 centres in 25 countries recruiting 4196 patients referred to a cardiologist due to CAD out of whom 2107 were admitted on an acute basis and 2854 had an elective consultation. Patient data were collected via a web-based case record form. An oral glucose tolerance test (OGTT) was used for the characterisation of the glucose metabolism. Thirty-one per cent of the patients had diabetes. An OGTT was performed on the 1920 patients without known diabetes, of whom 923 had acute and 997 had a stable manifestation of CAD, respectively. In patients with acute CAD, 36% had impaired glucose regulation and 22% newly detected diabetes. In the stable group these proportions were 37% and 14%. CONCLUSION: This survey demonstrates that normal glucose regulation is less common than abnormal glucose regulation in patients with CAD. OGTT easily discloses the glucometabolic state and should be a routine procedure. The knowledge of glucometabolic state among these patients should influence their future management because it has great potential to improve the outcome. PMID- 15522467 TI - Hyperinsulinaemia is associated with increased long-term mortality following acute myocardial infarction in non-diabetic patients. AB - AIMS: To study the impact of disturbances in glucose metabolism on total mortality in non-diabetic patients with acute myocardial infarction. METHODS AND RESULTS: Four hundred and ninety four patients with a verified myocardial infarction and no history of diabetes were studied. The study population comprised a subgroup of patients screened for participation in the Trandolapril Cardiac Evaluation (TRACE) study. At baseline, fasting insulin, fasting glucose, glycosylated haemoglobin (HbA1c), and urinary albumin excretion were measured. Survival status was determined after 6-8 years. Patients with hyperinsulinaemia were more obese and more frequently suffered from hypertension, previous myocardial infarction and congestive heart failure. In a univariate regression analysis, values in the upper quartile of insulin, glucose, HbA1c, and urinary albumin were associated with an excess mortality risk (RR=1.8 (1.2-2.7), p=0.002; RR=1.6 (1.2-2.1), p=0.001; RR= 1.9 (1.3-2.9), p=0.001; RR=1.6 (1.2-2.1), p=0.02 respectively). However, only a high insulin level remained significant in a multivariable analysis (RR=1.54 (1.03-2.31), p=0.04) including baseline variables, left ventricular systolic function and in-hospital complications. CONCLUSIONS: High fasting plasma insulin is an independent risk factor of all cause mortality in non-diabetic patients with acute myocardial infarction. This justifies future intervention studies aiming at reducing insulin resistance and using fasting insulin as the target variable. PMID- 15522468 TI - Antiplatelet effects of a 600 mg loading dose of clopidogrel are not attenuated in patients receiving atorvastatin or simvastatin for at least 4 weeks prior to coronary artery stenting. AB - AIMS: To test prospectively whether the antiplatelet effect of a 600 mg loading dose of clopidogrel is attenuated in patients receiving atorvastatin and simvastatin for at least 4 weeks prior to coronary artery stenting. METHODS AND RESULTS: Blood samples were obtained at least 2 h after receiving 100 mg aspirin and 600 mg clopidogrel and prior to coronary stenting from 90 patients without statin therapy and 90 patients with statin (atorvastatin and simvastatin) therapy for at least 4 weeks. Maximal and residual platelet aggregation was evaluated with optical aggregometry in response to ADP (5 and 20 micromol/l). Surface expression of IIb/IIIa (CD61) and P-selectin (CD62) was assessed with whole blood flow-cytometry at baseline and following stimulation (5 and 20 micromol/l ADP). Inhibition of ADP-induced platelet aggregation was not impaired in the presence of concomitant statin therapy. Moreover, patients with and without statin therapy did not differ in respect to all flow-cytometric parameters obtained. CONCLUSION: The antiplatelet effect of a high, 600 mg loading dose of clopidogrel is not diminished in patients receiving atorvastatin and simvastatin for at least 4 weeks prior to coronary stenting. PMID- 15522469 TI - High clopidogrel loading dose during coronary stenting: effects on drug response and interindividual variability. AB - AIM: To assess platelet inhibitory effects, interindividual variability in platelet inhibition as well as response to a 600 mg, compared to a standard 300 mg, clopidogrel loading dose (LD) after coronary stenting METHODS AND RESULTS: Platelet function profiles were assessed in 50 patients undergoing coronary stenting receiving either a 300 mg (n=27) or 600 mg clopidogrel LD. ADP (6 microM) and collagen (6 microg/mL) induced platelet aggregation, as well as ADP (2 microM) induced glycoprotein (GP) IIb/IIIa activation and P-selectin expression were assessed at baseline and 4, 24, and 48 h following clopidogrel front-loading. A more intense and rapid inhibition of platelet activation (both GP IIb/IIIa activation and P-selectin expression) were achieved using a 600 mg, compared to a 300 mg, LD throughout the entire 48 hours (p<0.001). Although there were no differences in platelet aggregation, overall a 600 mg LD increased the number of clopidogrel responders and this was also achieved earlier compared to a 300 mg LD. A 600 mg LD did not reduce interindividual variability of platelet response. CONCLUSION: The use of a 600 mg clopidogrel LD in patients undergoing coronary stenting optimises platelet inhibitory effects early after intervention and may provide a more effective protection against early thrombotic complications. PMID- 15522470 TI - An international perspective on heart failure and left ventricular systolic dysfunction complicating myocardial infarction: the VALIANT registry. AB - AIMS: We analysed the contemporary incidence, outcomes, and predictors of heart failure (HF) and/or left ventricular systolic dysfunction (LVSD) before discharge in patients with acute myocardial infarction (MI). The baseline presence of HF or LVSD, or its development during hospitalisation, increases short- and long-term risk after MI, yet its incidence, predictors, and outcomes have not been well described in a large, international, general MI population. METHODS AND RESULTS: The VALIANT registry included 5573 consecutive MI patients at 84 hospitals in nine countries from 1999 to 2001. A multivariable logistic survival model was constructed using baseline variables to determine the adjusted mortality risk for those with in-hospital HF and/or LVSD. Baseline variables were also tested for associations with in-hospital HF and/or LVSD. Of the 5566 patients analysed, 42% had HF and/or LVSD during hospitalisation. Their in-hospital mortality rate was 13.0% compared with 2.3% for those without HF and/or without LVSD. After adjustment for other baseline risk factors, in-hospital HF and/or LVSD carried a hazard ratio for in-hospital mortality of 4.12 (95% confidence interval: 3.08 5.56). Patients with HF and/or LVSD also had disproportionately higher rates of other cardiovascular events. CONCLUSIONS: HF and/or LVSD is common in the general contemporary MI population and precedes 80.3% of all in-hospital deaths after MI. Survivors of early MI-associated HF and/or LVSD have more complications, longer hospitalisations, and are more likely to die during hospitalisation. Although baseline variables can identify MI patients at highest risk for HF and/or LVSD, such patients are less likely to receive indicated procedures and medical therapies. PMID- 15522471 TI - Effect of candesartan on New York Heart Association functional class. Results of the Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity (CHARM) programme. AB - AIMS: To evaluate the effect of the angiotensin receptor blocker candesartan on New York Heart Association (NYHA) functional class in a broad spectrum of patients with chronic heart failure (CHF). METHODS AND RESULTS: Patients in the CHARM Programme with symptomatic CHF were randomized to placebo (n=3796) or candesartan (n=3803) and followed for a median of 38 months. NYHA class was assessed at baseline, at two weekly intervals during dose titration and 4 monthly thereafter. Patients were classified as "better", "unchanged" or "worse" at the end of the study compared to baseline. Both a simple "last visit carried forward" (LVCF) analysis and "worst rank carried forward" (WRCF) analysis (where patients who died were allocated NYHA class V) were used. In the LVCF analysis, compared to placebo, more candesartan patients improved (35.4% versus 32.5%) and fewer worsened (9.0% versus 10.3%) in NYHA class (p=0.003). The WRCF analysis also showed a better overall change in NYHA class with candesartan compared to placebo. There was no heterogeneity in the response to candesartan between the CHARM component trials. CONCLUSIONS: Candesartan improves NYHA functional class to a similar extent to other proven treatments for CHF when added to these other treatments. PMID- 15522472 TI - Doppler flow evaluation can anticipate abnormal left lung perfusion after transcatheter closure of patent ductus arteriosus. AB - AIMS: Coil protrusion into the left pulmonary artery (LPA) has been described after transcatheter closure of the patent ductus arteriosus (PDA). The possible impact of such a finding in lung perfusion has not been completely clarified. We evaluated Doppler flow velocities and lung perfusion in patients submitted to that procedure. METHODS: After transcatheter closure of PDA with coils, 70 patients (mean age 8.6+/-3.4 years) were followed for a period of 3.6+/-0.9 years (range 2.1-5.9) and compared to 22 controls. Peak flow velocities and coil protrusion were assessed by Doppler echocardiography. A Doppler velocity index (DVI) was calculated by the difference between the LPA and right pulmonary artery (RPA) peak flow velocities relative to the pulmonary trunk (PT) expressed in percentage, as follows: DVI=(LPA velocity - RPA velocity)/PT velocity x 100. Lung scintigraphy was performed using (99m)Tc-labelled macro-aggregated albumin. RESULTS: Device protrusion was observed in 94% of the patients, 10% of whom presented abnormal left lung perfusion. Peak LPA velocity and DVI were significantly greater in patients (p=0.001) and correlated negatively with left lung perfusion values (R(2)=0.21 and R(2)=0.65, respectively). A cut-off value of 50% for the DVI showed high sensitivity and specificity for reduced lung perfusion. CONCLUSION: Impaired left lung perfusion may appear following transcatheter closure of PDA with coils and the determination of DVI may anticipate such alteration. PMID- 15522473 TI - Tissue Doppler imaging detects early asymptomatic myocardial abnormalities in a dog model of Duchenne's cardiomyopathy. AB - AIMS: Early diagnosis of Duchenne's dilated cardiomyopathy remains a challenge for conventional echocardiography. We sought to determine whether tissue Doppler imaging (TDI) could detect early alteration in myocardial function in a dog model of Duchenne muscular dystrophy, i.e. the Golden Retriever Muscular Dystrophy (GRMD). METHODS AND RESULTS: Myocardial function was assessed by TDI in 20 dogs with normal conventional parameters of systolic function (eight controls and 12 GRMD, 25+/-11 weeks) without knowledge of the genotype. M-mode TDI was recorded from a short-axis view for measurement of endocardial and epicardial velocities and myocardial velocity gradient (MVG) within the posterior wall. Controls and GRMD dogs were comparable regarding left ventricular fractional shortening (37+/ 2 vs 42+/-3%, p=ns). Conversely, TDI showed, in all GRMD dogs, a dramatic decrease in systolic MVG (0.8+/-0.1 vs 2.9+/-0.3 s(-1), p<0.0001) and early diastolic MVG (2.3+/-2.2 vs 10.8+/-1.1 s(-1), p<0.0001). This MVG alteration was related to a significant decrease in endocardial velocities in GRMD whereas epicardial velocities were comparable in the two groups. CONCLUSION: These results show that TDI is more sensitive than conventional echocardiography in detecting pre-clinical myocardial abnormalities before occurrence of left ventricular dilation and dysfunction. TDI should be part of the screening techniques for the early diagnosis of cardiomyopathy. PMID- 15522474 TI - Clinical indications for cardiovascular magnetic resonance (CMR): Consensus Panel report. PMID- 15522475 TI - Relationship between reduced elasticity of extracardiac vessels and left main stem coronary artery disease. PMID- 15522476 TI - Metoprolol reduces the coronary collateral flow index: alternative explanations. PMID- 15522477 TI - Plasma B-type natriuretic peptide is related to left ventricular mass in hypertensive patients. PMID- 15522478 TI - Prognosis of heart failure in idiopathic dilated cardiomyopathy: has it improved in tertiary referral centres over the last 10 years? PMID- 15522479 TI - Equity, capabilities and health. AB - This paper provides an introduction to a collection of articles concerning the relevance of Amartya Sen's work, the capability approach and related ethical claims, to health-care rationing. PMID- 15522480 TI - The nature of procedural preferences for health-care rationing decisions. AB - Substantial debate on the appropriate foundations of economic evaluation in health-care has been conducted between welfarists and non-welfarists in the health economics literature. This has focussed on defining and measuring appropriate outcomes. However, there has been little discussion of the importance of procedures. This paper examines six dimensions of procedure in relation to health-care rationing which are drawn from existing literature and previous empirical investigation. A survey of the general public was used to test for preferences for each dimension of procedure. Results show that for each dimension the importance differed according to the level of decision-making (we tested decision-making at the level of the individual doctor, health authority and government). We identified three distinct clusters of respondents who can be broadly described as "proceduralists" (47%), "pluralists" (15%) and "anti consequentialists" (38%). The paper concludes that consequentialism is insufficient to provide the conceptual framework that public decision-making in health requires, although this does not mean that consequences are unimportant. PMID- 15522481 TI - Acceptability of less than perfect health states. AB - Health normally deteriorates beyond a certain age. This means, in Amartya Sen's terms, that one's health capabilities decline beyond a certain age, making it more difficult to achieve functionings such as mobility or sexual activity. In this paper, we investigate whether this normal reduction in quality of life also induces less than perfect health states to be considered acceptable at advanced stages of life. In other words, we investigate whether it is considered acceptable that health capabilities decline over time. In this study, we use domain-specific descriptions of health (mostly following the EQ-5D domains) in order to investigate whether the acceptability of less than perfect health states is similar for all types of health losses. Besides a theoretical consideration of this issue, we present some empirical evidence based on the answers of 226 respondents to a web-based survey. The results show that often individuals do indeed consider less than perfect health states acceptable, especially at more advanced stages of life. Mild health problems are more often considered acceptable than severe health problems. The acceptability of health states is related to the quality of life score of these states, i.e., worse states are considered less acceptable. This may have implications for the allocation of scarce health care resources. PMID- 15522482 TI - Communitarian claims and community capabilities: furthering priority setting? AB - Priority setting in health care is generally not done well. This paper draws on ideas from Amartya Sen and Martha Nussbaum and adds some communitarian underpinnings to provide a way of improving on current uses of program budgeting and marginal analysis (PBMA) in priority setting. The paper suggests that shifting to a communitarian base for priority setting alters the distribution of property rights over health service decision making and increases the probability that recommendations from PBMA exercises will be implemented. The approach is built on a paradigm which departs from three tenets of welfarism as it is normally conceived: (i) individuals qua individuals seek to maximise their individual utility/well-being; (ii) individuals want to do this; and (iii) it is the values of individuals qua individuals that count. Some of the problems of PBMA, as it has been applied to date, are highlighted. It is argued that these are due largely to a lack of 'credible commitment'. Bringing in the community and communitarian values to PBMA priority setting exercises can help to overcome some of the barriers to getting PBMA recommendations implemented. The approach has the merit of reflecting Sen's concept of capabilities (but extending that to a community level). It avoids the often consequentialist base of a conventional welfarist framework, and it allows community values as opposed to individual values to come to the fore. How to elicit communitarian values is explored. PMID- 15522483 TI - Concerns for the worse off: fair innings versus severity. AB - The original fair innings argument is about claims on length of life. Alan Williams has suggested that the argument also should apply to quality of life. His 'generalised fair innings approach' on the one hand, and the severity approach on the other, are two ways of incorporating concerns for fairness in economic evaluation of health care. They are based on different ethical arguments and therefore partly lead to different results. Both approaches incorporate concerns for current and future severity. There is strong support for this in formal theories of justice and government guidelines, and a number of public surveys even indicate the strength of these concerns. The generalised fair innings approach additionally incorporates concerns for past suffering. Intuitively, this is not unreasonable, but there is at this point little ethical theory or empirical evidence to suggest the strength of such concerns. The fair innings argument can be decomposed in an 'equal innings argument' and a 'sufficient innings argument'. When the fair innings argument is applied to quality of life, its sufficient innings component implies that young people should have priority over old people when it comes to functional improvements and symptom relief for non-fatal conditions. This runs counter to both moral intuitions and official goverment guidelines in Norway and Sweden. PMID- 15522484 TI - Empiricism, ethics and orthodox economic theory: what is the appropriate basis for decision-making in the health sector? AB - Economics is commonly defined in terms of the relationship between people's unlimited wants and society's scarce resources. The definition implies a central role for an understanding of what people want, i.e. their objectives. This, in turn, suggests an important role for both empirical research into people's objectives and debate about the acceptability of the objectives. In contrast with this expectation, economics has avoided these issues by the adoption of an orthodoxy that imposes objectives. However evidence suggests, at least in the health sector, that people do not have the simple objectives assumed by economic theory. Amartya Sen has advocated a shift from a focus on "utility" to a focus on "capabilities" and "functionings" as a way of overcoming the shortcomings of welfarism. However, the practicality of Sen's account is threatened by the range of possible "functionings", by the lack of guidance about how they should be weighted, and by suspicions that they do not capture the full range of objectives people appear to value. We argue that "empirical ethics", an emerging approach in the health sector, provides important lessons on overcoming these problems. Moreover, it is an ethically defensible methodology, and yields practical results that can assist policy makers in the allocation of resources. PMID- 15522485 TI - A "fair innings" between the sexes: are men being treated inequitably? AB - In most modern developed communities, women are known to live longer than men. A less known fact is that in many statistics reporting differences in life expectancy between socio-economic classes, on average women in the worst-off social class live as long as men in the best-off social class, if not longer. It is true that women tend to have higher morbidity, or lower health-related quality of life, especially at advanced age, but this female disadvantage does not offset the life expectancy advantage sufficiently to lead to the conclusion that men and women enjoy comparable lifetime health prospects in these communities. Although there is much public and policy discussion about the inequity of health inequalities between the social classes, there is relatively little discussion about such inequalities between the sexes. The paper first discusses the applicability of the fair innings argument to the issue of inequality in health between the sexes. It critically examines six arguments concerning why inequality in health between the sexes may or may not be an inequity. Next, special attention is given to the argument that it is wrong to judge the fairness or unfairness of health inequalities in isolation, but that this judgement should be made only after considering other inequalities relevant to overall human well being, many of which are believed to work to the disadvantage of women. An analytical framework based on the Gender-related Development Index (a supplementary index to the Human Development Index) is taken as a starting point, to address the issue of health and overall well-being. But this is found wanting, and suggestions are made as to how its conceptual and empirical properties could be improved. Meanwhile we conclude that a prima facie case has been made that the current distribution of health in most countries does not give men a "fair innings", but the broader question about general well-being remains unresolved. PMID- 15522486 TI - Understanding pregnancy in a population of inner-city women in New Orleans- results of qualitative research. AB - Unintended pregnancy has conventionally been defined as a pregnancy that is mistimed or unwanted, and this classification has been widely used in survey research. This study explores the utility of these constructs for women who visited a family planning clinic and a prenatal clinic in inner-city New Orleans, LA, and, by extension, for women of similar background and experience. We used semi-structured, open-ended research to explore sexual debut and history, contraceptive knowledge and use, pregnancy history, partner relations, and service use among 77 women (73 of whom were African-American). This study addresses the apparent paradox of high-risk sexual and contraceptive behavior in the presence of expressed preferences to postpone childbearing. It provides some insight into the cultural and social context in which these events and decisions take place and explores the multiple dimensions that shape women's sexual behaviors and their desires for pregnancy. The dimensions explored include perceptions of and experiences with sex/sexuality, values concerning childbearing/motherhood, relationships with partners, experiences with contraception, and attitudes toward abortion. The apparent ambivalence seen in reports of women asked whether a pregnancy was intended, such as statements that they did not want to get pregnant but were either not using contraception or using it irregularly, calls into question the idea that intendedness can be routinely and easily inferred from survey research. Correspondingly, it is not possible to simply assume that either intentionality or future intentions directly affect decisions to use contraception. The problem is that the many factors-structural and individual-affect women's preferences and ability to postpone a pregnancy or to use contraception. PMID- 15522487 TI - Are rich people or poor people more likely to be ill? Lay perceptions, by social class and neighbourhood, of inequalities in health. AB - Research in the UK has suggested that people in lower social classes or from poorer neighbourhoods are less likely than their more socially advantaged counterparts to agree that health and life expectancy are worse among more deprived population groups. The small body of previous research has either used qualitative approaches or coded open-ended responses to survey questions about causes of health and illness or of inequalities between areas. We examined lay perceptions by asking a direct question and using a quantitative, multivariate approach. Residents in three age groups (25, 45 and 65 years old) living in two socially contrasting localities in Glasgow, Scotland, were asked who were more likely to have accidents, cancer, heart disease, mental illness, to be fitter, and to live longer: rich people, poor people, or both equally. Across all the health categories, those in lower social classes or from poorer neighbourhoods were equally or less likely than their more socially advantaged counterparts to say the poor had worse health. In a model containing age, sex, class and locality, those in lower social classes and in the poorer locality were significantly less likely to say that richer people live longer (OR: 0.5). We have therefore confirmed earlier observations that those most at risk of ill health may be less likely to acknowledge the social gradient in health. We suggest a need to examine this apparent paradox in other contexts and in more detail, using both quantitative and qualitative approaches. PMID- 15522488 TI - Microbicide acceptability research: current approaches and future directions. AB - With growing recognition of the potential value of microbicides for HIV/STI prevention, the importance of the acceptability of this brand-new technology has been widely acknowledged. We review the current body of microbicide acceptability research, characterize the limitations in assessment approaches, and suggest strategies for improvement. Electronic databases and abstracts of recent meetings were searched for acceptability data regarding vaginal and rectal products that may be used for HIV prevention. Of the 61 studies reviewed, more than half assessed acceptability based primarily on the description of a hypothetical microbicide, or with the demonstration of a spermicide or lubricant. Physical characteristics of microbicidal products, their effects after insertion, and their effects on sensation during intercourse (for both partners) were the dimensions most frequently assessed (measured in 77%, 49% and 49% of studies, respectively). Attention to the social context of use was inadequate. As acceptability is likely to be a key determinant in the use-effectiveness of microbicides, in-depth understanding of the social processes that shape microbicide acceptability across diverse populations will become increasingly valuable. This includes exploring the effects that sexual partners, health care providers, and key opinion leaders have on the acceptability of microbicides among women and men, including youth and people living with HIV. Future research will benefit from studies of the acceptability of other contraceptive-barrier methods (especially the female condom), use of an agreed-upon operationalization of acceptability, use of acceptability assessments within clinical trials, expansion of measurement domains, and assessment of changes in perceptions of acceptability and use over time. Failure to understand the key factors associated with microbicide acceptability is likely to hinder the adoption and continued use of products that are effective in preventing HIV infection. PMID- 15522489 TI - Warranted concerns, warranted outlooks: a focus group study of public understandings of genetic research. AB - This paper discusses how the American public accounts for the concerns that they have about genetic research and the benefits that they foresee. We develop a general framework for discussing public claims about genetic technology based on Stephen Toulmin's model of warrants in argumentation. After a review of the results from public opinion polls about genetic research, we present a focus group study of public understandings of genetics. We outline the warrants, or publicly accepted "good reasons", that this group offers for accepting some aspects of genetic technology and for rejecting other aspects. The warrants presented by the public in their discussion of genetic research indicate that the public has a complex, informed understanding of genetic research, albeit a non technical one. The paper concludes with a discussion of the importance of public participation in debates over genetic research and the ways that researchers and policymakers could adapt to public concerns about genetics. PMID- 15522490 TI - Shifts in global immunisation goals (1984-2004): unfinished agendas and mixed results. AB - The turn of the millennium has been marked by a large-scale mobilisation of resources for immunisation programmes in developing countries. The resources have been generated by public and private sector parties collaborating in the Global Alliance for Vaccines and Immunization (GAVI). GAVI was formed in response to deteriorating immunisation coverage rates occurring in the late 1990s. GAVI is the latest in a line of vaccine initiatives, which have operated over the past 20 years. This article reviews the five most important global immunisation initiatives that have taken place over those past 20 years. It analyses their origins, shifts in global immunisation goals, identifies key actors, assesses the initiatives' capacity to mobilise resources and increase immunisation coverage, and points to possible unintended effects of the initiatives. The study argues that shifts in global immunisation goals lead to fragmentation in the implementation of vaccine programmes at the local level in developing countries. It also suggests that global actors involved in the formulation of these initiatives appear to miss opportunities to build on past experiences and fail to learn from previous mistakes. This raises questions about the initiatives' sustainability and relevance to the overall objective of preventing vaccine preventable deaths. PMID- 15522491 TI - Under what conditions is euthanasia acceptable to lay people and health professionals? AB - Euthanasia is legal only in the Netherlands and Belgium, but it is on occasion performed by physicians elsewhere. We recruited in France two convenience samples of 221 lay people and of 189 professionals (36 physicians, 92 nurses, 48 nurse's aides, and 13 psychologists) and asked them how acceptable it would be for a patient's physician to perform euthanasia in each of 72 scenarios. The scenarios were all combinations of three levels of the patient's life expectancy (3 days, 10 days, or 1 month), four levels of the patient's request for euthanasia (no request, unable to formulate a request because in a coma, some form of request, repeated formal requests), three of the family's attitude (do not uselessly prolong care, no opinion, try to keep the patient alive to the very end), and two of the patient's willingness to undergo organ donation (willing or not willing). We found that most lay people and health care professionals structure the factors in the patient scenarios in the same way: they assign most importance to the extent of requests for euthanasia by the patient and least importance (the lay people) or none (the health professionals) to the patient's willingness to donate organs. They also integrate the information from the different factors in the same way: the factors of patient request, patient life expectancy, and (for the lay people) organ donation are combined additively, and the family's attitude toward prolonging care interacts with patient request (playing a larger role when the patient can make no request). Thus we demonstrate a common cognitive foundation for future discussions, at the levels of both clinical care and public policy, of the conditions under which physician-performed euthanasia might be acceptable. PMID- 15522492 TI - The economics of social marketing: the case of mosquito nets in Tanzania. AB - There is a growing appreciation of the role of the private sector in expanding the use of key health interventions. At the policy level, this has raised questions about how public sector resources can best be used to encourage the private sector in order to achieve public health impact. Social marketing has increasingly been used to distribute public health products in developing countries. The Kilombero and Ulanga Insecticide-Treated Net Project (KINET) project used a social marketing approach in two districts of Tanzania to stimulate the development of the market for insecticide-treated mosquito nets (ITNs) for malaria control. Using evidence from household surveys, focus group discussions and a costing study in the intervention area and a control area, this paper examines two issues: (1) How does social marketing affect the market for ITNs, where this is described in terms of price and coverage levels; and (2) What does the added cost of social marketing "buy" in terms of coverage and equity, compared with an unassisted commercial sector model? It appears that supply improved in both areas, although there was a greater increase in supply in the intervention area. However, the main impact of social marketing on the market for nets was to shift demand in the intervention district, leading to a higher coverage market outcome. While social marketing was more costly per net distributed than the unassisted commercial sector, higher overall levels of coverage were achieved in the social marketing area together with higher coverage of the lowest socioeconomic group, of pregnant women and children under 5 years, and of those living on the periphery of their villages. These findings are interpreted in the context of Tanzania's national plan for scaling up ITNs. PMID- 15522493 TI - An explanatory model of health promotion and quality of life for persons with post-polio syndrome. AB - Although previous studies have examined selected factors influencing health promoting behaviors or quality of life, the complex interplay of these variables in persons with different chronic disabling conditions has not been investigated. This study tested an explanatory model of variables influencing health promotion and quality of life originally developed with a sample of persons with multiple sclerosis in a new sample of persons living with post-polio syndrome (PPS) in the USA. The sample of 1603 persons with PPS (1123 females, 478 males and 2 unknown) completed a battery of instruments including measures of severity of polio related impairment, barriers to health promoting behaviors, resources, self efficacy, acceptance, health promoting behaviors and perceived quality of life. A model originally developed in a sample of 786 persons with multiple sclerosis was assessed and modified using the weighted least squares procedure (WLS) which is implemented by LISREL8. The structural equation analyses resulted in a proper solution that exhibited adequate fit: chi2 (8, N = 1549)=84.22, p<0.05; GFI=0.96, IFI=0.90, CFI=0.90. The antecedent variables accounted for 65% of the variance in the frequency of health promoting behaviors and 53% of the variance in perceived quality of life. The model test supports the hypothesis that quality of life is the outcome of a complex interplay between contextual factors (severity of impairment), antecedent variables, and health promoting behaviors. It also suggests that the relationships among these variables are similar in samples of persons with two different chronic conditions. Further research using a qualitative approach is needed to clarify other contributors to quality of life in persons with post-polio syndrome. PMID- 15522494 TI - Maternal education and child nutritional status in Bolivia: finding the links. AB - This study models various pathways linking maternal education and child nutritional status in Bolivia, using a national sample of children. Pathways examined include socioeconomic status, health knowledge, modern attitudes towards health care, female autonomy, and reproductive behavior. The data come from the 1998 Bolivia Demographic and Health Survey. Logistic regression results suggest that socioeconomic factors are the most important pathways linking maternal education and child nutritional status, and that modern attitudes about health care also explain the impact of education. Health care knowledge accounts for less of the effect of maternal education on child nutritional status, with autonomy being the weakest pathway. Other pathways, such as reproductive behaviors, appear to influence nutritional status independent of maternal education. Overall, the pathways examined accounted for 60 percent of the effect of maternal education on child nutritional status. PMID- 15522495 TI - The causes of low back pain: a network analysis. AB - Beliefs regarding the cause of low back pain differ between individual sufferers and health care professionals. One consequence of this is the potential acquisition of maladaptive attitudes and behaviour in relation to pain, and increases in the utilisation of primary care services (Health Expect.3(3) (2000) 161). Methods that have been used to elicit the causal interpretation of social phenomena are varied yet they are unable to categorically demonstrate the different weightings or levels of importance that individuals may assign. The diagram method of network analysis allows individuals to spontaneously consider pathways they believe to be critical to a target event and to determine the strength of those pathways. Seventy-one completed diagrams indicating the causes that sufferers perceived to be related to low back pain were analysed. The mean number of direct causal paths was 5.61 (SD=3.25) and mean number of indirect causal links was 1.16 (SD=2.34). A significant correlation between path frequency and path strength was also found (r=0.76, p=0.001). Sufferers do not have an overtly complex view of the causative factors of low back pain but were able to define four core contributory causes (disc, sciatica, lifting, and injury) and one indirect pathway between lifting and injury. There was a clear delineation between external (biomedical) and internal (person-related) factors that were attributed to low back pain acquisition. By determining these causal attributions it is proposed that treatment packages could be tailored to address biases in thinking. This may be particularly useful for those individuals who attribute their pain as a consequence of external (or biomedical) causes. PMID- 15522496 TI - Perceived breast cancer risk: heuristic reasoning and search for a dominance structure. AB - Studies suggest that people construct their risk perceptions by using inferential rules called heuristics. The purpose of this study was to identify heuristics that influence perceived breast cancer risk. We examined 11 interviews from women of diverse ethnic/cultural backgrounds who were recruited from community settings. Narratives in which women elaborated about their own breast cancer risk were analyzed with Argument and Heuristic Reasoning Analysis methodology, which is based on applied logic. The availability, simulation, representativeness, affect, and perceived control heuristics, and search for a dominance structure were commonly used for making risk assessments. Risk assessments were based on experiences with an abnormal breast symptom, experiences with affected family members and friends, beliefs about living a healthy lifestyle, and trust in health providers. Assessment of the potential threat of a breast symptom was facilitated by the search for a dominance structure. Experiences with family members and friends were incorporated into risk assessments through the availability, simulation, representativeness, and affect heuristics. Mistrust in health providers led to an inappropriate dependence on the perceived control heuristic. Identified heuristics appear to create predictable biases and suggest that perceived breast cancer risk is based on common cognitive patterns. PMID- 15522497 TI - The health consciousness myth: implications of the near independence of major health behaviors in the North American population. AB - Analysis of over 250,000 respondents from four of the largest epidemiological surveys in North America indicates that major health behaviors are largely unrelated to one another. On average, the percentage of shared variance among smoking, exercise, diet and alcohol consumption is approximately 1%. While many of these relationships are statistically significant, suggesting that the associations are nonzero in the population, they represent minute effect sizes. The weak associations among these behaviors are unlikely to be due to incorrect functional form of the relationship, measurement error, or biases in responding. The findings have implications for health behavior theories and interventions predicated on the notion that the health conscious individual attempts to improve his or her health by engaging in more than one of these behaviors at a time. PMID- 15522498 TI - Variation in 4-mercapto-4-methyl-pentan-2-one release by Saccharomyces cerevisiae commercial wine strains. AB - The volatile thiol 4-mercapto-4-methylpentan-2-one (4MMP) is a potent contributor to wine aroma. In grape juice, 4MMP is bound to cysteine as a non-volatile compound and requires the action of yeast during fermentation to release the aroma active thiol. A method was developed to measure 4MMP release from the precursor by headspace solid-phase microextraction and separation by gas chromatography with atomic emission detection to screen the ability of wine yeast to release 4MMP. Yeast commonly used in white wine making were grown with the precursor at two different temperatures, and the amount of 4MMP released was measured. The results demonstrate that yeast strain selection and fermentation temperature can provide an important tool to enhance or modulate the grape derived aromas formed during wine fermentation. PMID- 15522499 TI - Anticancerogenic effect of a novel chiroinositol-containing polysaccharide from Bifidobacterium bifidum BGN4. AB - Strains of bifidobacteria have many health-promotion effects. Whole cells or cytoplasm extracts of Bifidobacterium bifidum BGN4, isolated from human feces, inhibited the growth of several cancer cell lines. The polysaccharide fraction (BB-pol) extracted from B. bifidum BGN4 had a novel composition, comprising chiroinositol, rhamnose, glucose, galactose, and ribose. Three human colon cancer cell lines were treated with BB-pol: HT-29, HCT-116, and Caco-2. Trypan blue exclusion assay and BrdU incorporation assay showed that BB-pol inhibited the growth of HT-29 and HCT-116 cells but did not inhibit the growth of Caco-2 cells. PMID- 15522500 TI - Beta-glucosidase activity from the thermophilic fungus Scytalidium thermophilum is stimulated by glucose and xylose. AB - An inducible mycelial beta-glucosidase from Scytalidum thermophilum was characterized. The enzyme exhibited a pI of 6.5, a carbohydrate content of 15%, and an apparent molecular mass of about 40 kDa. Optima of temperature and pH were 60 degrees C and 6.5, respectively. The enzyme was stable up to 1 h at 50 degrees C and exhibited a half-life of 20 min at 55 degrees C. The enzyme hydrolyzed p nitrophenyl-beta-d-glucopyranoside, p-nitrophenyl-beta-d-xylopyranoside, o nitrophenyl-beta-d-galactopyranoside, p-nitrophenyl-alpha-arabinopyranoside, cellobiose, laminaribiose and lactose. Kinetic studies indicated that the same enzyme hydrolyzed these substrates. Beta-Glucosidase was activated by glucose or xylose at concentration varying from 50 to 200 mM. The apparent affinity constants (K0.5) for glucose and xylose were 36.69 and 43.24 mM, respectively. The stimulatory effect of glucose and xylose on the S. thermophilum beta glucosidase is a novel characteristic which distinguish this enzyme from all other beta-glucosidases so far described. PMID- 15522501 TI - A divIVA null mutant of Staphylococcus aureus undergoes normal cell division. AB - DivIVA is involved in placement of the division septum and chromosome segregation in Bacillus subtilis and it plays important roles in cell division or morphogenesis in diverse Gram-positive bacteria. In Staphylococcus aureus, DivIVA is localized at the division septum, but it does not colocalize with the chromosomal origin of replication, as labeled with SpoOJ protein. Unexpectedly, a divIVA null mutant is not impaired in growth, nor is it affected in chromosome segregation or cell morphology. PMID- 15522502 TI - Release of transforming plasmid DNA from actively growing genetically engineered Escherichia coli. AB - We studied the transforming ability of the extracellular plasmid DNA released from a genetically engineered Escherichia coli pEGFP and the culturing conditions for the release of transforming DNA. The transforming ability was evaluated by transformation of competent cells with filtrates of E. coli pEGFP cultures. The number of transformants increased with time when E. coli pEGFP cells grew exponentially in rich medium, but not in stationary phase or when inoculated in freshwater. These results suggested that crude extracellular plasmid DNA had transforming ability and this transforming DNA was mainly released by actively growing bacteria. PMID- 15522503 TI - Molecular hydrogen from water radiolysis as an energy source for bacterial growth in a basin containing irradiating waste. AB - Although being deionized, filtered and therefore normally deeply oligotrophic, the water from a basin containing irradiating waste presented relatively high bacterial concentrations (ca 10(5) cfu ml(-1)) and biofilm development at its surface and on the walls. This water was characterized by a high concentration of molecular H2 due to water radiolysis, while its electrochemical potential was around +400 mV due the presence of dissolved O2 and active oxygen compounds. This combination of H2 availability and of an oxidant environment is completely original and not described in nature. From surface and wall biofilms, we enumerated the autotrophic populations ( approximately 10(5) bacteria ml(-1)) able to grow in presence of H2 as energy source and CO2 as carbon source, and we isolated the most abundant ones among cultivable bacteria. They efficiently grew on a mineral medium, in the presence of H2, O2 and CO2, the presence of the three gases being indispensable. Two strains were selected and identified using their rrs gene sequence as Ralstonia sp. GGLH002 and Burkholderia sp. GGLH005. In pure culture and using isotope exchange between hydrogen and deuterium, we demonstrated that these strains are able to oxidize hydrogen as energy source, using oxygen as an electron acceptor, and to use carbon dioxide as carbon source. These chemoautotroph hydrogen-oxidizing bacteria probably represent the pioneer bacterial populations in this basin and could be primary producers in the bacterial community. PMID- 15522504 TI - Genetic diversity of small eukaryotes from the coastal waters of Nansha Islands in China. AB - Population structures and genetic diversity of the small eukaryotic plankton from the coastal waters of the Nansha Islands in China were investigated. Two genes libraries using 18S rDNA of the marine small eukaryotes were constituted, and 323 clones were identified within alveolates (more than 43%), acanthareas, viridiplantaes, and stramenopiles. Many novel clones were detected in the two libraries, including two groups of alveolates and two clades related to both acanthareas and polycystineas. Several sequences unrelated to any other known eukaryotes may represent early branches in the phylogenetic tree. Our results reveal that there is a high diversity and abundance of small eukaryotes in the marine regions of China. PMID- 15522505 TI - Characterization of DegU, a response regulator in Listeria monocytogenes, involved in regulation of motility and contributes to virulence. AB - The degU (lmo2515) gene encodes a putative response regulator in the food-borne pathogen Listeria monocytogenes. It has 63% amino acid identity to the DegU response regulator of Bacillus subtilis. We have characterized the degU gene product in L. monocytogenes EGD by generation of a deletion mutant. The DeltadegU mutant was found to be non-motile in motility plate assay and no flagellin was detected. The mutant was attenuated in challenge of mice. Northern blot analysis suggested that the degU gene product is a transcriptional activator of the flagellin gene, flaA, at 25 degrees C. However, the degU gene product had no influence on the transcription of prfA encoding the major virulence regulator, PrfA. The results indicate that the putative DegU response regulator is a pleiotropic regulator involved in expression of both motility at low temperature and in vivo virulence in mice. PMID- 15522506 TI - Enzymatic inactivation and reactivation of chloramphenicol by Mycobacterium tuberculosis and Mycobacterium bovis. AB - Mycobacterium tuberculosis and Mycobacterium bovis are inhibited by chloramphenicol. Chloramphenicol acetyltransferase (CAT) converts chloramphenicol to inactive diacetyl chloramphenicol, but a mycobacterial carboxylesterase hydrolyzes the diacetyl product to active chloramphenicol. The esterase activity was eliminated by proteinase K and heat treatment. Protein extracts of M. tuberculosis and M. bovis hydrolyzed four other ester substrates. cat was inserted into the chromosome of both M. tuberculosis and M. bovis resulting in a level of chloramphenicol resistance that could be used to select for transformants. CAT assays in the resistant strain of M. tuberculosis showed interference due to esterase activity. This interference could be eliminated with the addition of a heating step. PMID- 15522507 TI - Repetitive elements sequence (REP/ERIC)-PCR based genotyping of clinical and environmental strains of Yersinia enterocolitica biotype 1A reveal existence of limited number of clonal groups. AB - REP- and ERIC-PCR genotyping were used to assess genetic heterogeneity among 81 strains of Yersinia enterocolitica biotype 1A isolated from India, Germany, France and the USA. Although both gave comparable results, ERIC fingerprints discriminated the strains better. The rep- (REP and ERIC) PCR genotyping showed that strains having different serotypes produced identical rep-profiles indicating their limited genetic diversity. The concatenated dendrogram of REP- and ERIC-PCR fingerprints clustered the biotype 1A strains into two major groups. In each group, majority of the Indian, European and American strains exhibited similarities ranging from 85% to >95%. Similarity of rep-PCR fingerprints amongst strains isolated from widely separated geographical regions revealed existence of a limited number of clonal groups of Y. enterocolitica biotype 1A. The present study failed to reveal unequivocal relationships between rep-PCR genotypes and the source of isolation. However, the clinical serotype O:6,30-6,31 strains formed a tight cluster and the aquatic O:6,30-6,31 strains formed a yet another tight cluster. PMID- 15522508 TI - Multicellular life cycle of magnetotactic prokaryotes. AB - Most multicellular organisms, prokaryotes as well as animals, plants, and algae have a unicellular stage in their life cycle. Here, we describe an uncultured prokaryotic magnetotactic multicellular organism that reproduces by binary fission. It is multicellular in all the stages of its life cycle, and during most of the life cycle the cells organize into a hollow sphere formed by a functionally coordinated and polarized single-cell layer that grows by increasing the cell size. Subsequently, all the cells divide synchronously; the organism becomes elliptical, and separates into two equal spheres with a torsional movement in the equatorial plane. Unicellular bacteria similar to the cells that compose these organisms have not been found. Molecular biology analysis showed that all the organisms studied belong to a single genetic population phylogenetically related to many-celled magnetotactic prokaryotes in the delta sub-group of the proteobacteria. This appears to be the first report of a multicellular prokaryotic organism that proliferates by dividing into two equal multicellular organisms each similar to the parent one. PMID- 15522509 TI - Differences in gene expression of the ciliate Paramecium caudatum caused by endonuclear symbiosis with Holospora obtusa, revealed using differential display reverse transcribed PCR. AB - We identified six genes of Paramecium caudatum, which differentially expressed in Holospora obtusa-bearing and H. obtusa-free cells using differential display reverse transcribed PCR (DDRT-PCR). Northern blot analyses revealed that two of the genes, CA10-3 and CA20-2, were expressed extensively in the H. obtusa-free cell, while the other four, AS16-1, CS14, CS21 and CA17-1, were expressed more in the H. obtusa-bearing cell. Putative amino acid sequences of CA10-3, AS16-1 and CA17-1 showed high homologies with known genes related to intracellular signaling, transcription and aerobic metabolism. CS14 and CS21 also showed homologies with some genes whose products are still functionally unknown, but CA20-2 encoded a novel protein. We show in this study that H. obtusa alters multiple gene expression of the host after establishing endosymbiosis. PMID- 15522510 TI - Oligonucleotide microarray for identification of Bacillus anthracis based on intergenic transcribed spacers in ribosomal DNA. AB - We developed a DNA microarray for identification of Bacillus anthracis and other phylogenetic groupings within the "Bacillus cereus group". Nucleotide sequences of 16S-23S ribosomal DNA internal transcribed spacers containing genes for tRNA(Ile) from 52 B. anthracis strains were found to be identical to sequences from seven strains published previously and different from all other bacteria. When 42 oligonucleotide probes targeting polymorphic sites were immobilized on glass slides and hybridized to fluorescently labeled PCR amplification products, one or more mismatches could be discriminated in all but one cases. Hence, hybridization events were highly specific and identification of B. anthracis was straightforward. PMID- 15522511 TI - Rapid detection of Panton-Valentine leukocidin from clinical isolates of Staphylococcus aureus strains by real-time PCR. AB - To allow rapid identification of Panton-Valentine leukocidin (PVL)-producing Staphylococcus aureus strains, a real-time PCR assay for detection of PVL was developed. This assay is convenient, since it can be applied directly on bacterial suspensions and does not require previous DNA purification. Furthermore, the assay was found to be highly reproducible, robust and specific, since positive results were generated exclusively with PVL-positive S. aureus strains, and neither with PVL-negative strains nor staphylococci other than S. aureus. PMID- 15522512 TI - Single-strand conformational polymorphism analysis of the ribosomal internal transcribed spacer 1 for rapid species identification within the genus Pythium. AB - Single-strand conformational polymorphism (SSCP) of the ribosomal internal transcribed spacer 1 (ITS-1) was characterized for 58 isolates of Pythium, representing 41 species from the five groups of Plaats-Niterink. Thirty-one species each produced a distinct SSCP pattern. Three species produced more than one unique pattern, corresponding to morphological subgrouping. The remaining seven species produced three distinct patterns with two or three morphologically similar species sharing a pattern. A successful blind test with four samples and the identification of eight previously unknown isolates from irrigation water demonstrated the reliability of this technique for species identification. Each SSCP pattern was defined and described by the positions of the top and bottom bands and the number of bands in between, which allows laboratories to use this technique without need to access the type isolates of Pythium species. PMID- 15522513 TI - Single mutations of residues outside the active center of the xylanase Xys1 Delta from Streptomyces halstedii JM8 affect its activity. AB - Mutagenesis of the xylanase Xys1 of Streptomyces halstedii JM8 has been done by error prone PCR. Mutants with modified hydrolytic activity were isolated, the recombinant variant proteins purified and the catalytic activities of each one determined and compared with the wild type enzyme. Two of the isolated single point mutants, m1 (G133D) and m8 (N148D), showed 22-25% increase in specific activity towards xylan compared to wild type xylanase. Two other mutants, m5a (D175A) and m7 (T160A), showed a significant reduction in specific activity of 40 50% with respect to the wild type enzyme. These residues are mainly located in the beta alpha-loops of the xylanase, the region showing the main structural divergences within family 10 of xylanases. This study shows the usefulness of random mutagenesis to point out some key residues not directly involved in the active center, but in which mutation produces subtle structural rearrangements affecting the enzymatic function. PMID- 15522514 TI - Analysis of C-3 epimerization in (24R)-24,25-dihydroxyvitamin D3 catalyzed by hydroxysteroid dehydrogenase. AB - Studies on the C-3 epimerization in (24R)-24,25-dihydroxyvitamin D(3) [24R,25(OH)(2)D(3)] were performed using hydroxysteroid dehydrogenases (HSDs). 3 Epi-24R,25(OH)(2)D(3) was formed from 24R,25(OH)(2)D(3) by the catalysis of 3alpha- or beta-HSD. These HSDs also catalyzed the C-3 epimerization in 3-epi 24R,25(OH)(2)D(3) to form 24R,25(OH)(2)D(3). 24R,25(OH)(2)D(3) and its C-3 epimer were separated by inclusion high-performance liquid chromatography using gamma cyclodextrin (gamma-CD) as the mobile phase additive or a gamma-CD bonded chiral column. The product derived from the intermediate during the C-3 epimerization was isolated from the incubation specimens and identified as (7Z)-(24R)-24,25 dihydroxy-9,10-secocholesta-4,7,10(19)-trien-3-one by several instrumental analyses including (1)H-nuclear magnetic resonance spectrometry. The occurrence of this compound strongly proves that the formation of the C-3 epimer by HSD involves a dehydrogenation process. The present study suggests that HSDs may catalyze the C-3 epimerization of vitamin D compounds and modulate their concentrations and biological activities in animals and humans. PMID- 15522515 TI - Optimization and validation of a micellar electrokinetic chromatographic method for the analysis of florfenicol. AB - We have optimized a micellar electrokinetic capillary chromatographic method for the separation of florfenicol and florfenicol amine, its degradation product. The separation was carried out using a 50mM sodium borate buffer (pH 9.0) containing 25mM of sodium dodecyl sulphate. The method selectivity was proven by the simultaneous separation of florfenicol and two structural antibiotics, chloramphenicol and thiamphenicol. The same system can also be applied for the quantitative determination of these antibiotics. The method was then validated regarding linearity, precision and accuracy. PMID- 15522516 TI - Analytical characterization of heparin by capillary zone electrophoresis with conductivity detection and polymeric buffer additives. AB - A capillary zone electrophoresis (CZE) method for the analytical characterization of intact (high-molecular-weight) heparin was developed. For the first time, a hydrodynamically closed CZE separation system with conductivity detector was used for the separation, detection and quantitation of this highly sulfated, linear polysaccharide. Glycine (25mM) adjusted to pH 9.0 by bis-Tris-propane served as the running electrolyte system. Polymeric additives, polyvinylpyrrolidone (PVP), dextran (DEX), were used to improve the separation selectivity as they strongly retarded the heparin macromolecule while they did not practically influence comigrating inorganic anions. The proposed electrophoretic method was successfully validated. It was convenient for the sensitive, simple, rapid and reproducible assay of heparin in raw materials and isotonic saline. Here, the use of the conductivity detector was advantageous as it allowed heparin to be analyzed without a sample pretreatment. The CZE method should be an alternative to the pharmacopoeial conventional gel electrophoresis having used in the quality control of heparin so far. In addition, it should be convenient to quantitative estimation of heparin present in a preparation used, e.g., as the chiral selector in CE separations. PMID- 15522517 TI - Automated robotic liquid handling/laser-based nephelometry system for high throughput measurement of kinetic aqueous solubility. AB - The ability to rapidly and consistently measure aqueous solubility in a preclinical environment is critical to the successful identification of promising discovery compounds. The advantage of an early solubility screen is timely attrition of compounds likely to fail due to poor absorption or low bioavailability before more costly screens are performed. However, due to the large number of compounds and limited sample amounts, thermodynamic solubility measurements are not feasible at this stage. A kinetic solubility measurement is an alternative to thermodynamic measurements at the discovery stage that provides a rank listing of solubility values with minimal sample requirements. A kinetic solubility measurement is attractive from an automation vantage because it features rapid data acquisition and is amenable to multi-well formats. We describe the use of a robotic liquid/plate handler coupled to nephelometry detection for the measurement of kinetic solubility. We highlight the liquid handling validation, serial dilution parameters, and a comparison to the previous method. Experiments to further enhance throughput, or increase confidence in the automation steps, are described and the effects of these experiments are presented. In our integrated nephelometry method, we observe rapid liquid handling with an error of less than 10%, after a series of validation studies, and a sample throughput up to 1800 compounds per week. We compare the nephelometry method with our semi-thermodynamic flow-injection analysis (FIA) method, and find a 75% bin agreement between the methods. PMID- 15522518 TI - A method for the direct analysis of drug compounds in plasma using a single restricted access material (RAM) column. AB - We describe an automated approach to analyzing whole plasma samples using online extraction without the need for an analytical column. A single restricted access material (RAM) column provided online extraction and pre-concentration of analytes while effectively removing proteins, salts and other biological materials found in the plasma sample matrix. The reduction in the plasma matrix enabled direct elution of the analytes from the extraction column to the mass spectrometer for selective detection. The precision of the method was evaluated using a proprietary therapeutic agent (Compound A) and was less than 5% over the range of 1-500ng/ml in spiked whole plasma, with an LOQ of 1ng/ml. A side-by-side comparison of RAM results from a pharmacokinetic study in rats was made with a traditional protein precipitation LC-MS method and a correlation of 0.993 was obtained between both methods. The injection-to-injection cycle time for the RAM method was 8min. Further automation was demonstrated by addition and mixing of the internal standard to all samples via an injection program of the autosampler. PMID- 15522519 TI - The characterisation of selected drugs with amine-containing side chains using electrospray ionisation and ion trap mass spectrometry and their determination by HPLC-ESI-MS. AB - The electrospray ionisation-ion-trap mass spectrometry (ESI-MS(n)) of selected drug compounds with amine-containing side chains has been investigated. Certain characteristic in-source fragmentations have been observed for these molecules. Sequential product ion fragmentation experiments (MS(n)) have been performed in order to elucidate the degradation pathways for the [M + H](+) ions and their predominant fragment ions. These MS(n) experiments also show certain characteristic fragmentations with respect to the amine-containing side chains. QTOF-MS/MS has been used to support the identity of the proposed fragments. The data presented in this paper therefore provides useful information on the structure of these compounds with amine-containing side chains and can be used in the characterisation of such drugs, their structurally related metabolites and unknown molecules of pharmaceutical significance extracted from animal and plant sources, for example. Amphetamine, clenbuterol, flurazepam and methadone can be identified and determined in mixtures at low ng/ml concentrations by the application of HPLC-ESI-MS which can also be used for their analysis in saliva samples. PMID- 15522520 TI - Superheated water chromatography-nuclear magnetic resonance spectroscopy and mass spectrometry of vitamins. AB - The water-soluble vitamins, pyridoxine, riboflavin, and thiamine, were separated by reversed-phase liquid chromatography using hot or superheated water as the mobile phase and were detected using a range of detectors, including ultraviolet and fluorescence spectroscopy and mass spectrometry. By using deuterium oxide as the eluent, direct on-line nuclear magnetic resonance spectra could be obtained with minimal spectral interference from the mobile phase. Some of the compounds showed deuterium exchange of alkyl-protons when separated at high temperatures. PMID- 15522521 TI - Molecularly imprinted solid phase extraction-pulsed elution-mass spectrometry for determination of cephalexin and alpha-aminocephalosporin antibiotics in human serum. AB - A highly selective molecularly imprinted solid phase extraction (MISPE)-pulsed elution (PE) method coupled with electrospray mass spectrometry (MS) was developed for the rapid screening and determination of cephalexin in alpha aminocephalosporin antibiotics. This method involved the solid phase extraction of cephalexin using a molecularly imprinted polymer micro-column, and pulsed elution with 1% trifluoroacetic acid in methanol, which contains sulindac as an internal standard for enhanced precision in MS detection. An LC/MS spectrometer was operated in the positive electrospray mode, and the selected-ion-recording (SIR) function was employed to detect the molecular ions of cephalexin, cefradine, cefadroxil and sulindac at m/z 348, 350, 363 and 357. Linearity was achieved in the cephalexin concentration range from 0.3 to 25microg/ml (or 5 500ng) (R(2) = 0.998). The detection limit was estimated at 0.04microg/ml (or 0.8ng) of cephalexin. Advantages of the newly developed MISPE-PE-MS, over the previously reported MISPE-DPE-FPE-UV, were evidenced in terms of detection limit, analysis time, solvent consumption, and simplicity of method development. PMID- 15522522 TI - Sensitive chemiluminescence assay for risperidone in pharmaceutical preparations. AB - A sensitive chemiluminescence (CL) method to determine the antipsychotic risperidone (RSP) is proposed, based on the catalytic effect of risperidone on the CL reaction between luminol and hydrogen peroxide in flow system. The increment of CL intensity was correlated with risperidone concentration in the range of 10pg ml(-1) to 1.0ng ml(-1) with a relative standard deviation of less than 5.0% (n = 5); and a limit of detection of 4pg ml(-1) (3sigma). At a flow rate of 2.0ml min(-1), the flow injection CL method exhibited both high sensitivity and excellent selectivity giving a throughput of 120 samples per hour. The proposed method was applied successfully to the determination of risperidone in pharmaceutical preparations. PMID- 15522523 TI - The effect of preprocessing methods in reducing interfering variability from near infrared measurements of creams. AB - This work is part of a study in which the possibility of NIR combined with some chemometrical methods is investigated as a suitable technique to classify clinical study samples of a cream. In this study, the influence of different preprocessing methods on the removal of spectral variations due to some variance sources has been investigated. The applied preprocessing methods are standard normal variate (SNV), detrend correction, offset correction, and first and second derivation. The investigated variance sources are different batches of ingredients, different samples of the same batch, different days and different positions of the sample cup in the sample drawer of the instrument. A nested ANOVA design has been applied in order to quantify the variances introduced by these variance sources. Since ANOVA is a univariate technique, the necessary variable (wavelength) selection has been performed by the Fisher criterion. The best results, i.e. largest reduction of interfering variability and clearest distinction between different clinical study samples, are obtained with the second derivative spectra. PMID- 15522524 TI - Simple, sensitive and rapid LC-MS/MS method for the quantitation of cerivastatin in human plasma--application to pharmacokinetic studies. AB - A simple and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for estimation of cerivastatin (I) in human plasma, a potent hydroxy-methylglutaryl-coenzyme A reductase inhibitor. The analyte and internal standard (atorvastatin, II) were extracted by liquid/liquid extraction with diethyl ether/dichloromethane (70/30, v/v). The chromatographic separation was performed on reverse phase Xterra ODS column with a mobile phase of water/acetonitrile (30/70, v/v) with 0.03% formic acid. The protonated analyte was quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The mass transitions m/z 460.4 --> 356.3 and 559.2 --> 440.3 were used to measure I and II, respectively. The lower limit of quantitation was 10pg/mL with a relative standard deviation of less than 15%. Acceptable precision and accuracy were obtained for concentrations over the calibration curve ranges (0.01-10ng/mL). Sample analysis time of 2min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The assay can be used to analyze human plasma samples to support phase I and II clinical studies. PMID- 15522525 TI - Development and validation of an assay for citric acid/citrate and phosphate in pharmaceutical dosage forms using ion chromatography with suppressed conductivity detection. AB - The paper describes the development and validation of a simple, rapid, accurate, and sensitive ion chromatographic procedure to assay total citrate (citric acid/citrate) and phosphate in nine dosage forms. The dosage forms chosen represent all dosage forms in USP27-NF22 for which the respective monographs require an assay for either citric acid/citrate or citric acid/citrate and phosphate. Citrate and phosphate were separated in <10min by a hydroxide selective column using anion-exchange chromatography with a 20mM potassium hydroxide eluent and detected by suppressed conductivity. The method showed linear responses over the concentration ranges 0.2-100microg ml(-1) (r(2) > 0.9990) for citrate and 0.2-60microg ml(-1) (r(2) = 0.9999) for phosphate, with limits of quantitation (signal-to-noise (S/N) = 10) of 0.2microg ml(-1) for both analytes. The accuracy of the procedure, determined by spiked recovery measurements, was within 95-105%. The intraday and the interday precision were demonstrated by the relative standard deviations (R.S.D.) of <1 and <2%, respectively, for both analytes. The ruggedness was determined by a full factorial design using analyst, equipment, column lot, and eluent preparation procedure as variables. The results show an overall R.S.D. of <3% and that an electrolytically generated 20mM KOH eluent produces assay results equivalent to a manually prepared 20mM NaOH eluent. PMID- 15522526 TI - Analysis of undeclared synthetic phosphodiesterase-5 inhibitors in dietary supplements and herbal matrices by LC-ESI-MS and LC-UV. AB - A liquid chromatography-electrospray ionisation-mass spectrometry (LC-ESI-MS) method was developed to screen for the presence of synthetic phosphodiesterase type 5 (PDE-5) inhibitors including sildenafil, tadalafil and vardenafil. The method was applied to the analysis of dietary supplements and bulk herbal materials. Bulk powders or composites of tablets, capsules or liquids were prepared and an extraction of PDE-5 inhibitors was performed using a mixture of acetonitrile and water with sonication. Identification of sildenafil, vardenafil or tadalafil was accomplished using a single quadrupole mass spectrometer coupled to a liquid chromatograph with an electrospray interface. Positive ion detection in the full scan mode was used while in-source collision induced dissociation (CID) provided several structurally significant fragment ions to aid in the mass spectral identification. Approximately half of the 40 botanical products analyzed were found to contain undeclared synthetic PDE-5 inhibitors. For products found to contain one of these three compounds by LC-MS, HPLC with UV detection was used for quantitation. PMID- 15522527 TI - Development and validation of a high-performance liquid chromatography assay and a capillary electrophoresis assay for the analysis of adenosine and the degradation product adenine in infusions. AB - A high-performance liquid chromatography (HPLC) method and a capillary electrophoresis (CE) method for the analysis of adenosine and the degradation product adenine in infusion solutions have been developed and validated. The HPLC separation of the analytes was achieved on a RP-18 column, using a mobile phase, consisting of 20mM ammonium acetate, pH 6.0, containing 5% of acetonitrile at a flow rate of 1ml/min. Thymidine was used as internal standard. The CE separation was performed in a fused-silica capillary with a 100mM sodium phosphate buffer, pH 2.7, at an applied voltage of 25kV, using cytidine as internal standard. The assays were validated with regard to linearity, range, limit of detection (LOD), limit of quantitation (LOQ), specificity, and precision. Both methods were specific allowing reliable quantification of the analytes. Compared to the CE method, HPLC analysis yielded a two- to five-fold lower LOD. With respect to analysis time, CE was faster than HPLC. The applicability of both methods for the determination of the purity and stability of adenosine in the infusion solutions is demonstrated. PMID- 15522528 TI - Simultaneous determination of levodopa and benserazide by stopped-flow injection analysis and three-way multivariate calibration of kinetic-spectrophotometric data. AB - The simultaneous determination of levodopa and benserazide in pharmaceutical formulations is described, based on the application of multidimensional partial least-squares regression to the kinetic-spectrophotometric data provided by diode array detection within a stopped-flow injection method where analytes react with periodate. Flow injection parameters were adequately optimized. Accurate analysis is performed with no sample pre-treatment steps, and with minimum experimental effort. Satisfactory recovery results were obtained on a number of synthetic and commercial samples, in the latter case including the comparison with liquid chromatography measurements. PMID- 15522529 TI - Automated simultaneous triple dissolution profiles of two drugs, sulphamethoxazole-trimethoprim and hydrochlorothiazide-captopril in solid oral dosage forms by a multicommutation flow-assembly and derivative spectrophotometry. AB - This article deals with the simultaneous determination of three dissolution profiles with the aid of the new and emerging continuous-flow methodology known as multicommutation. This methodology is based on a flow network of a set of solenoid valves controlled by the computer and acting as independent multicommutators to allow the easy and automated control of flowing solutions. The obtained three dissolution profiles from one dosage form are the whole formulation profile or "global profile" recommended by pharmacopoeias, and, at same time, are recorded two "individual" profiles from two drugs present in the formulation. This is the second attempt to obtain simultaneously three dissolution profiles with a single spectrophotometric detector and the first with the multicommutation methodology. The selected pharmaceutical formulations contained a couple of active principles with overlapped spectra, namely sulphamethoxazole and trimethoprim or hydrochlorothiazide and captopril. The obtained empirical plots profiles fitted with the Higuchi equation also known as the three-parameter equation. PMID- 15522530 TI - Characterization of the preferred orientation of delta-mannitol crystallites in tablets. AB - The crystallographic texture, i.e. the preferred orientation of crystallites of delta-mannitol samples, has been experimentally determined by pole figure analysis. The pole figures were measured with an X-ray diffraction texture goniometer. It was found that already the uncompressed delta-mannitol powder sample was slightly texturized so that the (0 2 0) plane was parallel to the upper surface of the sample. The degree of preferred orientation was found to significantly increase when the powder was compressed to a tablet with the minimum (74MPa) compression pressure. Nevertheless, the direction of the texture remained parallel to the tablet surface. Maximum compression (740MPa) did not increase the degree of preferred orientation further. The compression time (0 or 60s) was not found to noticeably affect the strength or direction of the texture. The extent of the texture inside the tablet was determined with a tablet surface grinding experiment. The degree of preferred orientation was found to decrease under the surface while the orientation remained the same. The results were confirmed with orientation distribution function (ODF) calculations. PMID- 15522531 TI - A liquid chromatographic method for simultaneous determination of amoxicillin sodium and sulbactam sodium in a combination formulation. AB - An isocratic liquid chromatographic method with UV detection at 210nm is described for simultaneous determination of amoxicillin sodium and sulbactam sodium in a new combination formulation. Chromatographic separation of the two drugs was achieved on a Hypersil C(18) column using a mobile phase consisting of a binary mixture of methanol and 0.01mol/l sodium acetate (5:95, v/v). The commonly used paired-ion aqueous mobile phase for the determination of penicillins was avoided in this study. The developed LC method offers symmetric peak shape, good resolution and reasonable retention time for both drugs. Linearity, accuracy and precision were found to be acceptable over the concentration ranges of 155.3-1553.0microg/ml for amoxicillin sodium and 45.0 450.0microg/ml for sulbactam sodium. The proposed LC method can be used for the quality control of formulated products containing these two drugs. PMID- 15522532 TI - Solubility and phase separation of benzocaine and salicylic acid in 1,4-dioxane water mixtures at several temperatures. AB - The solubilities of benzocaine and salicylic acid were determined in water dioxane mixtures at several temperatures (5-40 degrees C for benzocaine and 10-40 degrees C for salicylic acid). The solubility curves as a function of dioxane ratio showed a maximum at 90% dioxane at all temperatures. Above 25 degrees C, the homogeneous mixture splits into two liquid immiscible phases. For benzocaine, the initial dioxane concentration range at which phase separation takes place increased with temperature (50-60% at 25 degrees C, 50-70% at 30-35 degrees C and 40-70% at 40 degrees C). For salicylic acid, the dioxane concentration required for phase separation (40-60% dioxane) did not change with temperature. Phase separation was not related to solid phase changes (polymorphism or solvates). The phase composition and drug extraction at the drug-rich phase were determined. The apparent enthalpies of the solution process were a nonlinear function of the dioxane ratio for both drugs. The apparent enthalpy of solution of benzocaine was larger than that expected at the upper limit of phase separation (70% dioxane), whereas for salicylic acid the apparent enthalpy of solution decreased abruptly at the region corresponding to phase separation (40-70% dioxane). Both drugs showed a nonlinear pattern of enthalpy-entropy compensation. PMID- 15522534 TI - Flow injection-chemiluminescence determination of puerarin in pharmaceutical preparations. AB - Strong chemiluminescence was observed when cerium(IV) reacted with rhodamine 6G in sulfuric acid medium in the presence of puerarin. This phenomenon has been utilized to design a sensitive and selective flow injection-chemiluminescence method for the determination of puerarin. Under the optimum conditions, the proposed procedure has a linear range between 1.3 x 10(-9) and 8.0 x 10(-7)g/mL, with a detection limit of 8.4 x 10(-10)g/mL puerarin and a relative standard deviation of 1.86% (n = 11) at 5.0 x 10(-8)g/mL puerarin. The method was successfully applied to the determination of puerarin in pharmaceutical preparations. The mechanism of this chemiluminescence reaction has been proposed. PMID- 15522533 TI - Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--Part I. AB - This paper is the first part of a summary report of a new commission of the Societe Francaise des Sciences et Techniques Pharmaceutiques (SFSTP). The main objective of this commission was the harmonization of approaches for the validation of quantitative analytical procedures. Indeed, the principle of the validation of theses procedures is today widely spread in all the domains of activities where measurements are made. Nevertheless, this simple question of acceptability or not of an analytical procedure for a given application, remains incompletely determined in several cases despite the various regulations relating to the good practices (GLP, GMP, ...) and other documents of normative character (ISO, ICH, FDA, ...). There are many official documents describing the criteria of validation to be tested, but they do not propose any experimental protocol and limit themselves most often to the general concepts. For those reasons, two previous SFSTP commissions elaborated validation guides to concretely help the industrial scientists in charge of drug development to apply those regulatory recommendations. If these two first guides widely contributed to the use and progress of analytical validations, they present, nevertheless, weaknesses regarding the conclusions of the performed statistical tests and the decisions to be made with respect to the acceptance limits defined by the use of an analytical procedure. The present paper proposes to review even the bases of the analytical validation for developing harmonized approach, by distinguishing notably the diagnosis rules and the decision rules. This latter rule is based on the use of the accuracy profile, uses the notion of total error and allows to simplify the approach of the validation of an analytical procedure while checking the associated risk to its usage. Thanks to this novel validation approach, it is possible to unambiguously demonstrate the fitness for purpose of a new method as stated in all regulatory documents. PMID- 15522535 TI - Acid catalysed degradation of some spiramycin derivatives found in the antibiotic bitespiramycin. AB - Bitespiramycin is a novel antibiotic containing a number of 4''-acylated spiramycin derivatives (isovalerylspiramycins I-III, butanoylspiramycin III, propanoylspiramycin III and acetylspiramycin III) as major components. These spiramycin derivatives are susceptible to degradation in acid solution. Liquid chromatography-ion trap mass spectrometry (LC/MS(n)) was used to study the degradation of these spiramycin derivatives in simulated gastric fluid at 37 degrees C. All derivatives degraded by first-order reactions for which rate constants (k) and half-lives (t(1/2)) were calculated. Acyl groups at position 3 had less effect on acid-stability of spiramycin derivatives than acyl groups at position 4''. The introduction of 4''-acyl groups enhanced the acid-stability of spiramycin derivatives and altered the degradation pathway in simulated gastric fluid such that loss of forosamine rather than loss of mycarose becomes the major degradation pathway. PMID- 15522536 TI - Degradation and configurational changes of thioridazine 2-sulfoxide. AB - Thioridazine (THD) is a phenothiazine neuroleptic drug used for the treatment of psychiatric disorders. After oral administration THD is extensively biotransformed to thioridazine 2-sulfone (THD 2-SO(2)), thioridazine 5-sulfoxide (THD 5-SO) and thioridazine 2-sulfoxide (THD 2-SO). THD 2-SO and THD 5-SO have two chiral centres and therefore exist as two diastereoisomeric pairs. The degradation and epimerization of THD 2-SO in human plasma, buffer and methanolic solutions were studied using an enantioselective HPLC method. The samples were prepared by liquid-liquid extraction with diethyl ether and the chiral resolution of the enantiomers was carried out on a Chiralpak AD column using a mobile phase consisting of hexane:ethanol:2-propanol (90:7:3, v/v/v) containing 0.2% diethylamine. The method was validated and used to study the degradation and epimerization under different conditions of incubation. Our results showed that both enantiomers were stable at varying temperatures, pH and ionic strengths; however, solubility problems were observed, mainly at pH 8.5. The influence of light on stability was studied using methanolic solutions and degradation and epimerization of the THD 2-SO enantiomers were observed under UV light of 366 and 254nm, respectively. PMID- 15522537 TI - Stereoselective analysis of carvedilol in human plasma using HPLC/MS/MS after chiral derivatization. AB - A relatively high-throughput high-performance liquid chromatography/tandem mass spectrometry (HPLC-MS/MS) method using a chiral derivatization reagent was developed for the quantitative determination of carvedilol enantiomers in human plasma. S-carvedilol and R-carvedilol are extracted from human plasma by protein precipitation using acetonitrile containing racemic [(2)H(5)]-carvedilol as an internal standard. Extracts are then derivatized with 2,3,4,6-tetra-O-acetyl-beta glucopyranosyl isothiocyanate (GITC) and analysed using HPLC-MS/MS with a TurboIonspray (TIS) interface and selected reaction monitoring. Using 150microL of plasma, the method was validated over a concentration range of 0.2-200ng/mL. The maximum within-run precision observed in a three run quality control was 8.2% for S-carvedilol and 6.7% for R-carvedilol, respectively. The maximum percentage bias observed at all quality control sample concentrations was 9.4% for S carvedilol and 11.6% for R-carvedilol, respectively. The HPLC-MS/MS method was also compared with a previously developed high-performance LC/fluorescence method by analysing 25 samples containing racemic carvedilol. Based on results obtained, these two methods were found to be equivalent. However, compared with LC/fluorescence method, HPLC-MS/MS method is more sensitive, uses less plasma, and also employs a less time-consuming sample preparation process. PMID- 15522538 TI - HPTLC determination of caffeine in stimulant herbal products and power drinks. AB - The caffeine content of selected herbal products and energy drinks available in the Saudi market was determined by HPTLC-UV densitometric analysis. Pre-coated HPTLC silica gel plates (20cm x 10cm) were used for the analysis. The solvent system consisted of ethyl acetate-methanol (85:15, v/v), and caffeine was detected at 275nm. The developed method was validated for specificity, repeatability (C.V. < 5%), recovery (98.90 +/- 3.46), and accuracy (99.84 +/- 2.87). The levels of caffeine were 4.76-13.29% (w/w) and 0.011-0.032% (w/v), for the herbal products and the energy drinks, respectively. PMID- 15522539 TI - Determination of methylxanthines in urine by liquid chromatography with diode array UV detection. AB - A liquid chromatography-diode array UV detection (LC-UVDAD) method for the simultaneous determination of four methylxanthines (caffeine, theobromine, paraxanthine and theophylline) is described. The chromatographic separation was achieved on a LC-18-DB column using 20:80 methanol:buffer (5mM citric acid adjusted to pH 5 with triethylamine) as mobile phase. The method has been applied to urine samples. The overall procedure had % recoveries ranging from 81.6 +/- 2.6 (theophylline) to 99.3 +/- 6.3 (theobromine). The within-day (n = 5) and between-days (n = 5 over 5 days) coefficients of variation in urine ranged from 2.9% (theophylline) to 3.4% (theobromine) and from 5.2% (theophylline) to 6.2% (theobromine). Estimated LOD and LOQ in urine ranged from 0.15microg/ml (theophylline) to 0.3microg/ml (theobromine) and from 0.8microg/ml (theophylline) to 1.2microg/ml (theobromine), respectively. Urine samples naturally contaminated with the target analytes were found. PMID- 15522540 TI - Simultaneous HPLC determination of ketoprofen and its degradation products in the presence of preservatives in pharmaceuticals. AB - A novel and quick high-performance liquid chromatography (HPLC) method with UV spectrophotometric detection was developed and validated for the determination of five compounds in topical gel. The described method is suitable for simultaneous determination of active component ketoprofen, two preservatives methylparaben and propylparaben and two degradation products of ketoprofen--3-acetylbenzophenone and 2-(3-carboxyphenyl) propionic acid--in a topical cream after long-term stability tests using ethylparaben as an internal standard. The chromatographic separation was performed on a 5microm Supelco Discovery C18 column (125mm x 4mm i.d., Sigma-Aldrich); the optimal mobile phase for separation of ketoprofen, methylparaben, propylparaben, degradation products 3-acetylbenzophenone and 2-(3 carboxyphenyl) propionic acid and ethylparaben as internal standard consists of a mixture of acetonitril, water and phosphate buffer pH 3.5 (40:58:2, v/v/v). At a flow rate of 1.0ml min(-1) and detection at 233nm, the total time of analysis was less than 10min. The method was applied for routine analysis (batch analysis and stability tests) of these compounds in topical pharmaceutical product. PMID- 15522541 TI - Voltammetric determination of pyridoxine (vitamin B6) by use of a chemically modified glassy carbon electrode. AB - A novel carbon nanotube-modified glassy carbon electrode was described for the direct determination of pyridoxine. The electrochemical behavior of pyridoxine was investigated, and a well-defined oxidation peak with high sensitivity was observed at the modified electrode. Owing to the unique structure and extraordinary properties of multi-wall carbon nanotube (MWNT), the MWNT-modified glassy carbon electrode shows obvious electrocatalytic activity to the oxidation of pyridoxine, since it greatly enhances the oxidation peak current of pyridoxine as well as lowers its oxidation overpotential. Based on this, a very sensitive and simple voltammetric method was developed for the measurement of pyridoxine. A sensitive linear voltammetric response for pyridoxine was obtained in the concentration range of 5 x 10(-7)-1 x 10(-4)mol/L, and the detection limit is 2 x 10(-7)mol/L using differential pulse voltammetry. Compared with other voltammetric methods, this proposed method possesses many advantages such as very low detection limit, fast response, low cost and simplicity. The practical application of this new analytical method was demonstrated with pyridoxine drugs. PMID- 15522542 TI - High-performance liquid chromatographic method for simultaneous determination of baicalein and baicalein 7-glucuronide in rat plasma. AB - A simple HPLC method with ultraviolet detection was developed for the simultaneous determination of baicalein and its major metabolite, baicalein 7 glucuronide (baicalin) in rat plasma. Following solid phase extraction with HLB cartridge, the analytes were separated using a gradient mobile phase consisting of methanol-acetonitrile-phosphate buffer. The flow-rate was set at 1ml/min and the eluent was detected at 320nm. The method is linear over the studied range of 1-10 and 0.05-1microg/ml for baicalein and baicalin, respectively. The intra-day and inter-day variations of the analysis were less than 6.38%, with relative error ranging from -3.6 to 6%. The limit of quantification in plasma was 0.05microg/ml for both baicalein and baicalin. The developed method proves to be an improved and more reliable method because of its capability for determining baicalein and baicalin simultaneously in a single chromatographic run. PMID- 15522543 TI - Interference of isonicotinyl hydrazone in the microbiological analysis of rifampicin from anti-tuberculosis FDC products containing isoniazid. AB - Microbiological assay is a sensitive method for the estimation of rifampicin (R). In the present study, interference due to isonicotinyl hydrazone (HYD), an interaction product of R and isoniazid (H), was checked during microbiological analysis of R, employing Bacillus subtilis and Sarcina lutea. The assays were done by disc diffusion method. Both R and HYD showed linear log response curves in the range of 0.01-10microg. In the presence of HYD, R was overestimated when tested against S. lutea and underestimated in case of B. subtilis. The same extent and type of interference was observed on assay of a marketed anti tuberculosis fixed-dose combination product, subjected to accelerated stability testing (40 degrees C/75% RH) for 1 month. This means that response of organisms used in microbiological assay of R might vary in the presence of HYD, with possibility of incorrect conclusions. Therefore, the study suggests that before a microbiological method involving a particular organism is extended to the determination of R in FDC formulations containing H, it should be tested for the influence of HYD and used only if non-interfering. PMID- 15522544 TI - HPLC determination of mycophenolic acid and mycophenolic acid glucuronide in human plasma with hybrid material. AB - Mycophenolic acid (MPA), the active metabolite of the prodrug mycophenolate mofetil is an immunosuppressive agent which inhibits inosine monophosphate dehydrogenase. MPA is metabolised to phenolic glucuronide (MPAG) that may be hydrolysed in vivo to form free MPA. Drug monitoring is required in patients with multi-organ failure. Here, we report a HPLC method with organic/inorganic hybrid material for the simultaneous analysis of MPA and MPAG in human plasma. MPA and MPAG and carboxy butoxy ether mycophenolic acid (MPAC) used as internal standard were analysed on a bonded X-Terra column with a linear gradient elution mode using orthophosphoric acid and acetonitrile as eluents. Sample treatment procedure consists of deproteinisation with acetonitrile. Analytical recoveries were higher than 98 and 89% at concentrations ranging from 1 to 25 and 20 to 200mg/L for MPA and MPAG, respectively. Calibration curves fitted by plotting the peak area ratio (compound of interest/internal standard) versus concentration were linear in the range 0.2-50mg/L for MPA and in the range 1-500mg/L for MPAG. The quantification limit was 0.2mg/L for MPA and 1mg/L for MPAG with a coefficient of variation less than 20% for a 500microL sample volume. Intra- and inter-assay coefficient of variation was lower than 7% for all compounds. Detection was performed at 215nm. Peak identity was confirmed through library matching by comparison with reference spectra. The X-Terra column provides good peak shape and may be used at low pH with a long life-time column. This HPLC method using a simple sample treatment procedure appears suitable for therapeutic drug monitoring in organ-transplant patients. The method is sensitive enough for monitoring MPA and MPAG during pharmacokinetic studies. PMID- 15522545 TI - Determination of cobalt in pharmaceutical products. AB - The aim of this paper is to determine the content of cobalt in pharmaceutical products (B(12) vitamin powder, B(12) ampoules, Centrum, Spectrum ABC and Optima Forte) by spectrometric (FAAS, GFAAS and ICP-AES) and electrometric (AdSV) analytical techniques. The samples (approximately 0.5g) were treated with a mixture of 6mL HNO(3) and 1mL H(2)O(2) in the microwave oven. Due to the matrix effects the method of standard addition is preferred. The validity of the methods was tested by recovery studies of standard addition and results were found to be satisfactory. PMID- 15522546 TI - Rapid quantitative analysis of oxiracetam in human plasma by liquid chromatography/electrospray tandem mass spectrometry. AB - A rapid and accurate reversed-phase liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the quantitative determination of oxiracetam in human plasma. Oxiracetam, a highly polar compound, was successfully retained by Atlantis dC18 reversed-phase column and detected with triple-quadrupole tandem mass spectrometry. After addition of internal standard (piracetam) to human plasma, plasma was simply precipitated with two volume of acetonitrile, evaporated and dissolved in 0.1% acetic acid. This method for the determination of oxiracetam was accurate and reproducible, with a limit of quantitation of 0.2microg/ml in human plasma. The standard calibration curve for oxiracetam was linear (r(2) = 0.999) over the concentration range 0.2-40.0microg/ml in human plasma. The intra- and inter-day precision over the concentration range of oxiracetam was lower than 8.3% (relative standard deviation, %R.S.D.), and accuracy was between 92.5 and 106.4%. PMID- 15522547 TI - Indirect photometric assay determination of gabapentin in bulk drug and capsules by capillary electrophoresis. AB - A capillary electrophoresis (CE) method was developed for the assay determination of gabapentin (GBP) in bulk drug and capsules. Separation was carried out on 74cm (62.5cm effective length) x 75microm i.d. fused silica capillary by applying a potential of -20kV at ambient temperature. Background electrolyte (BGE) consisting of 5mM 5-sulphosalicylic acid and 0.5mM cetyltrimethylammonium bromide (CTAB), pH 11.0 was employed for the separation. Indirect method of UV detection was performed at a wavelength of 215nm using sulphosalicylate ion as a chromophore. A linear calibration curve was obtained over a concentration range 20-200microg/ml of GBP in deionized water with a correlation coefficient (r) of 0.9998. Recoveries were shown to be >/=98% both in bulk drug and capsules with standard deviation (S.D.) zinc > nickel, which is the same as that determined for MT in solution. Thus, the MT chip can be an effective biosensor for the detection and measurement of several metal ions. PMID- 15522604 TI - Palladium hexacyanoferrate hydrogel as a novel and simple enzyme immobilization matrix for amperometric biosensors. AB - An amperometric glucose biosensor with glucose oxidase (GOx) immobilized into palladium hexacyanoferrate (PdHCF) hydrogel has been prepared and evaluated. The sensor was based on a two-layer configuration with biocatalytic and electrocatalytic layers separately deposited onto the electrode. To reduce the overpotential for reduction of hydrogen peroxide liberated in the enzyme catalyzed oxidation of glucose, an inner thin layer of nickel hexacyanoferrate (NiHCF) electrodeposited onto the surface of graphite electrode was used as an electrocatalyst. As an outer layer, the hydrogel of palladium hexacyanoferrate with entrapped glucose oxidase was used. Under optimal operating conditions (pH 5.0 and E = -0.075 V versus calomel (3.0 M KCl) reference electrode), sensor showed high sensitivity to glucose (0.3-1.0 microA/mM) and a response time of less than 30s. The linear response to glucose was obtained in the concentration range between 0.05 and 1.0 mM in batch analysis mode and 0-7.0 mM in FIA. During the 32 days testing period, no significant decrease in the sensor sensitivity was observed. The sensor was applied for the determination of glucose concentration in fruit juice and yoghurt drink, and the results obtained showed good correlation with results obtained by reference spectrophotometric enzyme method. PMID- 15522605 TI - An enzyme electrode for amperometric measurement of D-amino acid. AB - A carbon paste enzyme electrode has been developed for measurement of D-amino acids that employs a fatty acid modified FAD to prevent leaching of this essential cofactor to the surrounding aqueous environment and which serves as an enzyme stabilizing agent. The lower limit of detection is at least 10(-4) M and the electrode has a linear range of 10(-4) to 3 x 10(-3) M and a response time of 180 s. Twenty L-amino acids were tested and none of them elicited responses when electrodes were exposed to 0.5 mM concentration increases over a baseline level. On the other hand, some response was observed when exposed to 18 of 20 D-amino acids varying from 2 to 200% of the corresponding D-alanine response. Electrodes were shown to have longevities of over 30 days while maintaining 85% of their original sensitivity. Electrodes showed activity over a pH of 6.2-11.7 with a maximum at 9.2 and over temperatures of 10-47 degrees C with a maximum at 37 degrees C. PMID- 15522606 TI - Fabrication of comb interdigitated electrodes array (IDA) for a microbead-based electrochemical assay system. AB - This research is directed towards developing a more sensitive and rapid electrochemical sensor for enzyme labeled immunoassays by coupling redox cycling at interdigitated electrode arrays (IDA) with the enzyme label beta galactosidase. Coplanar and comb IDA electrodes with a 2.4 microm gap were fabricated and their redox cycling currents were measured. ANSYS was used to model steady state currents for electrodes with different geometries. Comb IDA electrodes enhanced the signal about three times more than the coplanar IDAs, which agreed with the results of the simulation. Magnetic microbead-based enzyme assay, as a typical example of biochemical detection, was done using the comb and coplanar IDAs. The enzymes could be placed close to the sensing electrodes (approximately 10 microm for the comb IDAs) and detection took less than 1 min with a limit of detection of 70 amol of beta-galactosidase. We conclude that faster and more sensitive assays can be achieved with the comb IDA. PMID- 15522607 TI - Detection of insulin-antibody binding on a solid surface using imaging ellipsometry. AB - Imaging ellipsometry (IE) was used to detect the binding of insulin to its antibody on a solid surface. The modification of a gold surface with 11 mecaptoundecanoic acid (11-MUA), the adsorption of protein G, and antibody immobilization onto the protein G layer were confirmed by surface plasmon resonance. Ellipsometric images and ellipsometric angles of the surface antibody were acquired using the IE system by off-null ellipsometry. Ellipsometric images of antigen binding to the antibody were acquired, and their mean optical intensities estimated. Changes in mean optical intensity indicated that the detection range for insulin was from 10 ng/ml to 100 microg/ml. PMID- 15522608 TI - Regeneration of ethyl parathion antibodies for repeated use in immunosensor: a study on dissociation of antigens from antibodies. AB - Reliable analysis using an immunosensor strongly depends on the specificity, activity, and sensitivity of the antibody. Immobilization of antibody on the solid matrix enables its repeated use, for which it is required to dissociate the antigens and antigen-enzyme conjugate from the immobilized antibody matrix after each use and while doing so, a maximum retention of activity and specificity are crucial requirements. In the present investigation, on the development of an immunosensor for the organophosphorus pesticide ethyl parathion (EP) using EP antibodies, different dissociating agents such as organic solvents, detergents and acidic buffers, that is, dimethyl sulphoxide (DMSO), Tween-20, cetyl trimethylammonium bromide (CTAB), methanol, chloroform, guanidium chloride (GdmCl), glycine-HCl (Gly-HCl) buffer in the pH range of 1.5-3.0, pierce buffer and combination of DMSO and methanol in phosphate buffer and Gly-HCl buffer and salts like NaCl and MgCl2 were used. Generally about 50-60% dissociation was obtained with some degree of denaturation of the antibody immobilized on the sepharose matrix. However, 1% DMSO in combination with 0.2 M Gly-HCl buffer at a pH of 2.3 showed 97% dissociation and the immobilized antibody retained sufficient activity to carry out 14 reproducible assays for EP. PMID- 15522609 TI - Study on the mechanism of Bioelectric Recognition Assay: evidence for immobilized cell membrane interactions with viral fragments. AB - The Bioelectric Recognition Assay (BERA) is a whole-cell based biosensing system that detects the electric response of cultured cells, suspended in a gel matrix, to various ligands, which bind to the cell and/or affect its physiology. Previous studies have demonstrated the potential application of this method for rapid, inexpensive detection of viruses in a crude sample. However, the understanding, so far, of the fundamental processes that take place during cell-virus interactions within the probe has been rather limited. In the present study, we combined electrophysiological and fluorescence microscopical assays, so that we can prove that animal and plant cells immobilized in BERA sensors respond to different viruses primarily by changing their membrane potential. The response of immobilized cells against different viruses did not depend on the virus ability to penetrate the cell, but was modified after binding each virus to a virus specific antibody or removal of its coat protein after treatment with a protease. Consequently, we were able to assay the presence of a virus in its complete form or fragments thereof. Combination of immunological recognition with the electrophysiological response of immobilized cells allows for a considerable increase of the specificity of the BERA biosensory assay. In addition, rather than simply detect the presence of a protein or genomic sequence, the method can help gain information on the bioactivity of a virus. PMID- 15522610 TI - Orcokinins in insects and other invertebrates. AB - Orcokinin (NFDEIDRSGFGFN) and orcokinin homologues are crustacean peptides eliciting potent myotropic effects in gut tissues. Through HPLC purification of brain extract of the cockroach Blattella germanica, we isolated the first insect orcokinin (NFDEIDRSGFNS). This insect orcokinin-like peptide do not show myotropic properties in B. germanica gut tissues. Gene database search using orcokinin precursor sequences of the crustacean Procambarus clarkii led to putative homologues found in non-crustacean groups, including the mosquito Anopheles gambiae and the nematode Caenorhabditis elegans. PMID- 15522611 TI - Alternative farnesoid structures induce different conformational outcomes upon the Drosophila ortholog of the retinoid X receptor, ultraspiracle. AB - In view of recent studies that the ligand-binding pocket of the Drosophila melanogaster nuclear hormone receptor, ultraspiracle (dUSP), is a necessary component of dUSP-dependent transcriptional activation by methyl epoxyfarnesoate, we have assessed qualitative differences in the effect of farnesoid and dodecanoid compounds on receptor conformation and transcriptional activation. Farnesoids possessing terminal alcohol, aldehyde, acid, ester and/or epoxide moieties induced different changes in the local environment of the ligand-binding pocket, as monitored by the change each induced in the fluorescence of the two tryptophan residues existing in dUSP (that are situated 10 residues apart on the alpha-helix 5 that forms one lining of ligand-binding pocket). Similarly, each compound differed in the extent that it promoted an increase in anisotropy (dimerization state) of the receptor. Dodecanoid derivatives were much weaker in causing such effects. Methyl expoxyfarnesoate (insect juvenile hormone III) exhibited the greatest biological activity to increase transcription of a DR12JHECore reporter construct in transfected Sf9 cells, even though it did not exert the most suppression of USP fluorescence nor exert the greatest increase in USP anisotropy. In a comparison of farnesoid derivatives possessing the three side branches either as all methyl groups (JH III), or one of the side branches as ethyl (JH II), or two of the side branches as ethyl (JH I), the JH III and JH I were more similar to each other in the fluorescence suppression and in vivo morphogenetic activity than either was to JH II, evidencing that dUSP does not sense JH II as a structural 'intermediate' between JH III and JH I. Ligand binding domains of vertebrate retinoid X receptors respond to agonists by repositioning alpha-helix 12 to the edge of a hydrophobic groove, and there with the groove jointly forms a coactivator binding surface. When alpha-helix 12 in dUSP was mutated to place two signaling tryptophan residues its C-terminus, fluorescence signaling indicated that upon dUSP binding of methyl epoxyfarnesoate, the alpha-helix 12 was repositioned differently than what occurred upon binding of non-JH farnesoids. These leads on alternative ligand induced conformations that dUSP can adopt provide a foundation for commercial development of synthetic molecules that induce specific dUSP conformations, and for identification of in vivo conditions under which endogenous molecules may exert these conformational outcomes to this receptor. PMID- 15522612 TI - TsetseEP, a gut protein from the tsetse Glossina morsitans, is related to a major surface glycoprotein of trypanosomes transmitted by the fly and to the products of a Drosophila gene family. AB - African trypanosomes live in the lumen of the gut of tsetse (Glossina) and may have to face an immune response. As yet, it is unclear whether they are sensitive to antimicrobial peptides in vivo, but for some years there has been indirect evidence that one or more lectins can influence the infection. We have purified a protein complex from midgut extracts that, by SDS-PAGE, is a doublet of 37 and 38 kDa in a ratio of 3:1. Through prediction from corresponding cDNA clones, the full-length protein (tsetseEP) contains 320 amino acids, including a signal peptide. There is apparently only one gene encoding this protein. Towards the C terminus, the protein contains a run of 59 (EP) repeats, which surprisingly is what comprises almost the entire mature EP procyclin molecule present on the surface of trypanosomes in the tsetse gut. Drosophila contains a number of genes encoding proteins, of unknown function, with the same cysteine pattern as tsetseEP; this pattern is not reported for any other protein. Immunoblotting with a monoclonal antibody against (EP) repeats reveals expression in the gut, but not salivary glands, of female and male flies, whether or not fed. Immunoelectron microscopy shows the presence in vesicles in midgut cells and in the lumen of the gut. Attempts to demonstrate lectin activity were thwarted by limited availability of the protein complex. PMID- 15522613 TI - Enhanced esterase gene expression and activity in a malathion-resistant strain of the tarnished plant bug, Lygus lineolaris. AB - Extensive use of insecticides on cotton in the mid-South has prompted resistance development in the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois). A field population of tarnished plant bugs in Mississippi with 11-fold higher resistance to malathion was used to examine how gene regulation conferred resistance to this organophosphate insecticide. In laboratory bioassays, synergism by the esterase inhibitors S,S,S,-tributylphosphorotrithioate (DEF) and triphenylphosphate (TPP) effectively abolished resistance and increased malathion toxicity by more than 80%. Esterase activities were compared in vitro between malathion susceptible and resistant (selected) strains. More than 6-, 3- and 10 fold higher activities were obtained with the resistant strain using alpha naphthyl acetate, beta-naphthyl acetate, and p-nitrophenyl acetate, respectively. Up to 95% and 89% of the esterase activity in the susceptible and resistant strains, respectively, was inhibited by 1 mM DEF. Inhibition of esterase activity up to 75% and 85% in the susceptible and resistant strains, respectively, was obtained with 0.03 mM TPP. Esterase activities in field populations increased by up to 5.4-fold during the fall season. The increase was synchronized with movement of the insect into cotton where exposure to pesticides occurred. Esterase cDNA was cloned and sequenced from both malathion susceptible and resistant strains. The 1818-nucleotide cDNA contained a 1710-bp open reading frame coding a 570 amino acid protein which was similar to many insect esterases conferring organophosphate resistance. No amino acid substitution was observed between susceptible and resistant strains, indicating that esterase gene mutation was not involved in resistance development in the resistant strain in Mississippi. Further examination of esterase gene expression levels using quantitative RT-PCR revealed that the resistant strain had a 5.1-fold higher level of esterase mRNA than the susceptible strain. The results of this study indicated that up-regulation of the esterase gene appeared to be related to the development of resistance in the tarnished plant bug. PMID- 15522614 TI - Molecular crowding as a mechanism for tick secretory granule biogenesis. AB - During feeding ticks secrete bioactive components into the host to counter-act its immune and hemostatic defense systems. These bioactive components are stored in secretory granules that are secreted during feeding in an exocrine stimulus response type of mechanism. All proteins destined for secretion are packaged into these granules during granule biogenesis. Up to date no mechanism for granule biogenesis has been proposed, except to note that biogenesis occurs under conditions of high protein and calcium concentrations in an acidic environment. Previously, the most abundant proteins (TSGPs) found in the salivary glands of the soft tick, Ornithodoros savignyi, were suggested to play a part in granule biogenesis, based on their high abundance. The TSGPs are part of the lipocalin family, of which numerous members have been identified in ticks. We consider here the high concentrations of the TSGPs in salivary glands and what effect this will have on the crowded environment inside the secretory granules. It is shown that the TSGPs occur at concentrations that will lead to molecular crowding of which one result is the non-specific aggregation of components to reduce crowding effects. Aggregation of proteins as a mechanism of granule biogenesis has been proposed before, but not in terms of molecular crowding. We thus propose molecular crowding as the general mechanism of granule biogenesis, in tick secretory granules, but can also be extended to other forms of secretory granules in general. PMID- 15522615 TI - Molecular characterization of a novel peroxidase involved in Aedes aegypti chorion protein crosslinking. AB - Peroxidase-catalyzed chorion or eggshell protein crosslinking is an important biochemical event contributing to the formation of a protective chorion or eggshell in insects. Although the survival of the developing or developed embryo in the environment before hatching relies on the protection of the chorion, the identity of the peroxidative enzyme responsible for mediating chorion protein crosslinking has never been identified in any insect species. In this report, we describe the determination of partial peptide sequences of a novel mosquito chorion peroxidase through LC/MS/MS of a trypsin-digested chorion peroxidase active fraction, specific localization of the enzyme in the chorion through histochemical analysis, proteolytic processing of chorion peroxidase through comparison of the accurate mass of its intact mature enzyme with molecular mass of its deduced amino acid sequence, isolation of its cDNA based on chorion peroxidase partial amino acid sequences, evaluation of its transcriptional profile in developing ovaries, and application of the primary mosquito chorion peroxidase sequence in predicting potential chorion peroxidases in other species. PMID- 15522616 TI - AcMNPV ChiA protein disrupts the peritrophic membrane and alters midgut physiology of Bombyx mori larvae. AB - Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) chitinase A (ChiA) is a protein which promotes the final liquefaction of infected host larvae. The potential of this viral molecule as a new tool for insect control is explored here. The ChiA gene was isolated from the AcMNPV genome by PCR and expressed in E. coli. The recombinant protein, purified by affinity chromatography, showed both exo- and endo-chitinase activities and produced perforations on the peritrophic membrane (PM) of Bombyx mori larvae which increased in number and in size, in a dose-dependent manner. This structural alteration resulted into a significant increase of PM permeability to methylene blue and to the small neuropeptide proctolin. When the fifth instar larvae of B. mori were fed on a artificial diet supplemented with the recombinant ChiA, 100% mortality was observed at a dose of 1 microg/g of larval body weight (LW), while at sub-lethal doses of 0.56 microg/g LW, a reduced larval growth was recorded. These results indicate that AcMNPV-ChiA may offer interesting new opportunities for pest control. PMID- 15522617 TI - Selectivity and neuroendocrine regulation of the precursor uptake by pheromone glands from hemolymph in geometrid female moths, which secrete epoxyalkenyl sex pheromones. AB - Macrolepidopteran female moths in families such as Geometridae produce epoxyalkenyl sex pheromones, which are biosynthesized via epoxidation of polyunsaturated hydrocarbons in their pheromone glands. The precursors, however, are expected to be produced outside of the pheromone glands, probably in oenocytes or in the fat body, and transported to the glands via hemolymph. Based on these facts, the selectivity of the epoxidation substrates and of the precursor uptake by pheromone glands was examined with two geometrid species, Hemerophila artilineata and Ascotis selenaria cretacea, using binary mixtures of deuterated precursors and their analogs, which were topically applied to the pheromone glands or injected into the abdomen. GC-MS measurements of pheromone extracts showed equal epoxidation of two polyenes, indicating a low selectivity for both processes, while the epoxidation proceeded at only one double bond specific to each species. This result makes it possible to conclude that the formation of species-specific epoxyalkenyl pheromones results from the rigid formation of polyunsaturated precursors and their epoxidation at a fixed position. Next, the neuroendocrine regulation of these processes was studied with in vivo and in vitro experiments using decapitated females. The epoxy pheromones disappeared completely within 36 h of decapitation, and epoxidation of the injected precursors was not detected in the decapitated females, which restarted the reaction by treatment with a pheromone biosynthesis-activating neuropeptide (PBAN). The precursors topically applied to glands of the decapitated females, however, were converted into epoxy pheromones without PBAN, indicating that this neuropeptide hormone accelerated the precursor uptake by pheromone glands but not the epoxidation already underway in the glands. PMID- 15522620 TI - Mechanisms of antigen receptor evolution. AB - The adaptive immune system, which utilizes RAG-mediated recombination to diversify immune receptors, arose in ancestors of the jawed vertebrates approximately 500 million years ago. Homologs of immunoglobulins (Igs), T cell antigen receptors (TCRs), major histocompatibility complex (MHC) I and II, and the recombination activating genes (RAGs) have been identified in all extant classes of jawed vertebrates; however, no definitive ortholog of any of these genes has been identified in jawless vertebrates or invertebrates. Although the identity of the "primoridal" receptor that likely was interrupted by the recombination mechanism in the common ancestor of jawed vertebrates may never be established, many different families of genes that exhibit predicted characteristics of such a receptor have been described both within and outside the jawed vertebrates. Various model systems point toward a range of immune receptor diversity, encompassing many different families of recognition molecules, including non-diversified and diversified Ig-type variable (V) regions, as well as diversified VJ domains, whose functions are integrated in an organism's response to pathogenic invasion. The transition from the primordial antigen receptor to the monomeric Ig-/TCR-like domain and subsequent antigen specific heterodimer likely involved progressive refinement of unique intermolecular associations in parallel with the acquisition of combinatorial diversity and antigen-specific recognition through somatic modification of the V region. RAG-mediated recombination and associated junctional diversification of both Ig and TCR genes occurs in all jawed vertebrates. In the case of Igs, somatic variation is expanded further through class switching, gene conversion, and somatic hypermutation. Various approaches, including both genomic and protein functional analyses, currently are being applied in jawless vertebrates, protochordates and other invertebrate deuterostome model systems in order to examine both RAG-mediated and alternative forms of antigen receptor diversification. Such studies have uncovered previously unknown mechanisms of generating receptor diversity. PMID- 15522618 TI - DNA-based detection of Bt resistance alleles in pink bollworm. AB - Evolution of resistance by pests is the main threat to long-term insect control by transgenic crops that produce Bacillus thuringiensis (Bt) toxins. We previously identified three mutant alleles (r1, r2, r3) of a cadherin gene in pink bollworm (Pectinophora gossypiella) linked with recessive resistance to Bt toxin Cry1Ac and survival on transgenic Bt cotton. Here we describe a polymerase chain reaction (PCR)-based method that detects the mutation in genomic DNA of each of the three resistant alleles. Using primers that distinguish between resistant and susceptible (s) alleles, this method enables identification of 10 genotypes (r1r1, r1r2, r1r3, r2r2, r2r3, r3r3, r1s, r2s, r3s, and ss) at the cadherin locus. For each of the three resistant alleles, the method detected the resistance allele in a single heterozygote (r1s, r2s, or r3s) pooled with DNA from the equivalent of 19 susceptible (ss) individuals. The results suggest that the DNA-based detection method described here could greatly increase the efficiency of monitoring for resistance to Cry1Ac compared to bioassays that detect rare individuals with homozygous resistance. PMID- 15522621 TI - Origins of lymphocyte developmental programs: transcription factor evidence. AB - This review explores the evolutionary origins of lymphocyte development by focusing on the transcription factors that direct mammalian lymphocyte development today. Gene expression data suggest that the programs to make lymphocytes involve the same transcription factor ensembles in all animals with lymphocytes. Most of these factors, GATA, Runx, PU.1/Spi, EBF/Olf, Ikaros, and Pax-2/5/8 family members, are also encoded in the genomes of animals without lymphocytes. We consider the functions of these factors in animals without lymphocytes in terms of discrete program components, which could have been assembled in a new way to create the lymphocyte developmental program approximately 500 My ago. PMID- 15522622 TI - The evolutionary and structural 'logic' of antigen receptor diversity. AB - Most vertebrate species utilize antibody and T-cell receptor (TCR) genes to create a vast repertoire of antigen sensor molecules on their B and T lymphocytes, respectively. While the organization of these genes exhibits substantial variation between species, one common theme is that, in almost every case, there is at least one variable region with a highly diverse CDR3 region and often much less diversity elsewhere in the binding site. Whereas with alphabeta TCRs this skewing of diversity correlates well with the need to recognize diverse peptides bound to MHC molecules, this cannot explain why this same pattern is evident in immunoglobulins (Igs) or gammadelta TCRs. Instead we have postulated that in the primary repertoire, all or most antigen receptors have a bipartite binding site, in which diverse CDR3 loops act as a highly antigen specific 'core' whereas other CDRs bind in a largely opportunistic fashion. In the case of antibodies, somatic hypermutation then acts to improve the complementarity to a given antigen and increase antibody affinity. A test of this model in mice engineered to have a very limited V region repertoire shows that primary antibodies can be generated that are highly specific for distinct antigens, yet identical in sequence except for their V(H) CDR3. Furthermore, very high affinity antibodies can be raised by repeated immunizations, showing that somatic hypermutation can mold these low affinity antibodies into high affinity ones. Thus, the wide variations seen in V region repertoires amongst vertebrates is likely to be of lesser importance than the preservation of one or more diverse CDR3 regions. PMID- 15522623 TI - Antigen receptor genes and the evolution of a recombinase. AB - Antigen receptor genes exist in the germline in a "split" configuration and are assembled in developing B and T lymphocytes by V(D)J recombination. This site specific recombination reaction is initiated by a complex containing the RAG1 and RAG2 proteins and completed by general DNA repair factors. RAG1 and RAG2, like the adaptive immune system itself, are found exclusively in jawed vertebrates, and are thought to have entered the vertebrate genome by horizontal transmission as components of a transposable element. This review discusses the structure of antigen receptor genes and the mechanisms by which they are assembled and diversified, and then goes on to consider the evolutionary implications of the arrival of the hypothetical "RAG transposon". PMID- 15522624 TI - Evolution of isotype switching. AB - This review discusses evolution of the process of Ig heavy chain class switching, relating it to the first appearance of somatic hypermutation (SHM) of variable region genes. First, we discuss recent findings on the mechanism of class switch recombination (CSR) in mice and humans, and then review the mechanisms of expression of Ig heavy chain isotypes from fishes to mammals. Importantly, activation-induced cytidine deaminase (AID), which is essential for CSR and somatic hypermutation, is found in fishes. Although at least some fishes are likely to undergo SHM, CSR is highly unlikely to occur in this group. We discuss the first appearance of CSR in amphibians and how it differs in birds and mammals. PMID- 15522625 TI - A bird's eye view on human B cells. AB - We show in this review that there is a continuum between the chicken B-cell system classified as the first GALT model described and the human B-cell system. We propose that humans have conserved for one B-cell subpopulation, the marginal zone B-cell subset in charge of T-independent responses, the strategies of diversification used by GALT species to generate their pre-immune repertoire. PMID- 15522626 TI - Subjective experience and the attentional lapse: task engagement and disengagement during sustained attention. AB - Three experiments investigated the relationship between subjective experience and attentional lapses during sustained attention. These experiments employed two measures of subjective experience (thought probes and questionnaires) to examine how differences in awareness correspond to variations in both task performance (reaction time and errors) and psycho-physiological measures (heart rate and galvanic skin response). This series of experiments examine these phenomena during the Sustained Attention to Response Task (SART, Robertson, Manly, Adrade, Baddeley, & Yiend, 1997). The results suggest we can dissociate between two components of subjective experience during sustained attention: (A) task unrelated thought which corresponds to an absent minded disengagement from the task and (B) a pre-occupation with one's task performance that seems to be best conceptualised as a strategic attempt to deploy attentional resources in response to a perception of environmental demands which exceed ones ability to perform the task. The implications of these findings for our understanding of how awareness is maintained on task relevant material during periods of sustained attention are discussed. PMID- 15522627 TI - Motion-induced blindness does not affect the formation of negative afterimages. AB - Aftereffects induced by invisible stimuli constitute a powerful tool to investigate what type of neural information processing can occur in the absence of visual awareness. This approach has been successfully used to demonstrate that awareness of oriented gratings or translating stimuli is not necessary to obtain a robust orientation-specific or motion-specific aftereffect. We exploit motion induced blindness (MIB, Bonneh, Cooperman, & Sagi, 2001) to investigate the related question of the influence of visual awareness on the formation of negative afterimages. Our results show that MIB does not affect the persistence and intensity of afterimages. Thus, there is no significant contribution to the formation of afterimages beyond the sites mediating MIB. PMID- 15522628 TI - Identifying neural correlates of consciousness: the state space approach. AB - This article sketches an idealized strategy for the identification of neural correlates of consciousness. The proposed strategy is based on a state space approach originating from the analysis of dynamical systems. The article then focuses on one constituent of consciousness, phenomenal awareness. Several rudimentary requirements for the identification of neural correlates of phenomenal awareness are suggested. These requirements are related to empirical data on selective attention, on completely intrinsic selection and on globally unconscious states. As an example, neuroscientific findings on synchronized gamma activity are categorized according to these requirements. PMID- 15522629 TI - Handedness and the fringe of consciousness: strong handers ruminate while mixed handers self-reflect. AB - Previous research found that mixed handers (i.e., those that are more ambidextrous) were more likely than strong handers to update their beliefs (Niebauer, Aselage, & Schutte, 2002). It was assumed that this was due to greater degrees of communication between the two cerebral hemispheres in mixed handers. Niebauer and Garvey (2004) made connections between this model of updating beliefs and metacognitive processing. The current work proposes that variations in interhemispheric interaction (as measured by degree of handedness) contribute to differences in consciousness, specifically when consciousness is used in rumination versus the metacognitive task of self-reflection. Using the Rumination Reflection Questionnaire (Trapnell & Campbell, 1999), predictions were supported such that strong handedness was associated with self-rumination; whereas, mixed handedness was associated with increased self-reflection p values<.01, (N=255). James's (1890) concept of the "fringe of consciousness" is used to make connections between metacognition, updating beliefs, and self-reflection. Several studies are reviewed suggesting that mixed handers experience fringe consciousness to a greater degree than strong handers. PMID- 15522630 TI - Neurocognitive mechanisms underlying the experience of flow. AB - Recent theoretical and empirical work in cognitive science and neuroscience is brought into contact with the concept of the flow experience. After a brief exposition of brain function, the explicit-implicit distinction is applied to the effortless information processing that is so characteristic of the flow state. The explicit system is associated with the higher cognitive functions of the frontal lobe and medial temporal lobe structures and has evolved to increase cognitive flexibility. In contrast, the implicit system is associated with the skill-based knowledge supported primarily by the basal ganglia and has the advantage of being more efficient. From the analysis of this flexibility/efficiency trade-off emerges a thesis that identifies the flow state as a period during which a highly practiced skill that is represented in the implicit system's knowledge base is implemented without interference from the explicit system. It is proposed that a necessary prerequisite to the experience of flow is a state of transient hypofrontality that enables the temporary suppression of the analytical and meta-conscious capacities of the explicit system. Examining sensory-motor integration skills that seem to typify flow such as athletic performance, writing, and free-jazz improvisation, the new framework clarifies how this concept relates to creativity and opens new avenues of research. PMID- 15522631 TI - On time, memory and dynamic form. AB - A common approach to explaining the perception of form is through the use of static features. The weakness of this approach points naturally to dynamic definitions of form. Considering dynamical form, however, leads inevitably to the need to explain how events are perceived as time-extended--a problem with primacy over that even of qualia. Optic flow models, energy models, models reliant on a rigidity constraint are examined. The reliance of these models on the instantaneous specification of form at an instant, t, or across a series of such instants forces the consideration of the primary memory supporting both the perception of time-extended events and the time-extension of consciousness. This cannot be reduced to an integration over space and time. The difficulty of defining the basis for this memory is highlighted in considerations of dynamic form in relation to scales of time. Ultimately, the possibility is raised that psychology must follow physics in a more profound approach to time and motion. PMID- 15522632 TI - The consequences of encoding information on the maintenance of internally generated images and thoughts: the role of meaning complexes. AB - Three experiments investigated the hypothesis that internally generated images and thoughts were driven by meaning complexes, a construct which reflects a synthesis of semantic meaning and personal salience (Klinger, 1999). Experiments 1 and 2 contrasted the mutual inhibition between encoding words and non-words on: (i) the frequency that thoughts and images unrelated to the task (task unrelated thought, TUT) were experienced (Experiment 1) and (ii) on the intensity of images generated from long-term memory and maintained under dual task conditions, which whilst familiar were not of particular personal salience (Experiment 2). Experiment 3 examined the physiological arousal associated with the experience of TUT in a semantic encoding task. Evidence suggested that, in general, internally generated images and thoughts, irrespective of the personal salience, were suppressed by the co-ordination of information in working memory. In addition, only the experience of spontaneous images and thoughts of personal salience (TUT, Experiments 1 and 3) interfered reliably with the encoding/retrieval of semantic information from memory. Finally, in Experiment 3, physiological arousal, as indexed by mean heart rate, was associated with a high frequency of TUT. The results of all three experiments support the notion that the maintenance of spontaneously occurring images and thoughts is simultaneously influenced by both the semantic content and the personal salience of the information held in working memory. PMID- 15522633 TI - Dissociative style and individual differences in verbal working memory span. AB - Dissociative style is mostly studied as a risk factor for dissociative pathology, but it may also reflect a fundamental characteristic of healthy information processing. Due to the close link between attention and working memory and the previous finding of enhanced attentional abilities with a high dissociative style, a positive relationship was also expected between dissociative style and verbal working memory span. In a sample of 119 psychology students, it was found that the verbal span of the high-dissociative group was about half a word larger than of the medium and low-dissociative groups. It is suggested that dissociative style may be one of only very few individual differences that is directly relevant to consciousness research. PMID- 15522634 TI - A comparison of masking by visual and transcranial magnetic stimulation: implications for the study of conscious and unconscious visual processing. AB - Visual stimuli as well as transcranial magnetic stimulation (TMS) can be used: (1) to suppress the visibility of a target and (2) to recover the visibility of a target that has been suppressed by another mask. Both types of stimulation thus provide useful methods for studying the microgenesis of object perception. We first review evidence of similarities between the processes by which a TMS mask and a visual mask can either suppress the visibility of targets or recover such suppressed visibility. However, we then also point out a significant difference that has important implications for the study of the time course of unconscious and conscious visual information processing and for theoretical accounts of the processes involved. We present evidence and arguments showing: (a) that visual masking techniques, by revealing more detailed aspects of target masking and target recovery, support a theoretical approach to visual masking and visual perception that must take into account activities in two separate neural channels or processing streams and, as a corollary, (b) that at the current stage of methodological sophistication visual masks, by acting in more highly specifiable ways on these pathways, provide information about the microgenesis of form perception not available with TMS masks. PMID- 15522635 TI - Phenomenal characteristics associated with projecting oneself back into the past and forward into the future: influence of valence and temporal distance. AB - As humans, we frequently engage in mental time travel, reliving past experiences and imagining possible future events. This study examined whether similar factors affect the subjective experience associated with remembering the past and imagining the future. Participants mentally "re-experienced" or "pre-experienced" positive and negative events that differed in their temporal distance from the present (close versus distant), and then rated the phenomenal characteristics (i.e., sensorial, contextual, and emotional details) associated with their representations. For both past and future, representations of positive events were associated with a greater feeling of re-experiencing (or pre-experiencing) than representations of negative events. In addition, representations of temporally close events (both past and future) contained more sensorial and contextual details, and generated a stronger feeling of re-experiencing (or pre experiencing) than representations of temporally distant events. It is suggested that the way we both remember our past and imagine our future is constrained by our current goals. PMID- 15522636 TI - Blood free morphine levels vary with concomitant alcohol and benzodiazepine use. AB - AIMS: To record the concentration of morphine in post-mortem blood samples from fatalities arising directly from illicit drug abuse in Grampian, Scotland and to explore whether there is any difference in the morphine levels with co-existent drugs especially alcohol and benzodiazepines. METHODS: Drug-related deaths that had positive blood toxicology for morphine from the beginning of January 1995 until the end of December 2001 were identified from the database of toxicological data held in the Department of Clinical Biochemistry, Aberdeen Royal Infirmary, and the data analysed in conjunction with the information in the files of the Forensic Medicine Unit, University of Aberdeen. RESULTS: One hundred and twenty six cases were identified and consisted of 106 males and 20 females with an age range 17-51 (Median 26 years). Morphine was the single isolate in only 21 cases giving a median blood free morphine (BFM) of 0.19 mg/L. When morphine and benzodiazepines were the primary isolates [Formula: see text] the median BFM was 0.37 mg/L, and when morphine was in combination with alcohol [Formula: see text] the median BFM was 0.18 mg/L. Statistical significance was noted between these groups ( [Formula: see text] ). The remaining 59 cases had morphine combined with other drugs. CONCLUSIONS: In the studied cases, the concentrations of BFM were statistically higher when morphine was only isolated in conjunction with benzodiazepines than when combined only with alcohol. These results may be multi factorial; user error while under drug influence, the individuals drug tolerance, or related to the pharmacokinetics of morphine and its interactions with alcohol and benzodiazepines. Our understanding of the interactions of these substances appears to be far from clear. PMID- 15522637 TI - A study of suicides in Kildare, 1995-2002. AB - This paper focuses on 109 cases of suicide that occurred in Kildare from 1995 to 2002. These statistics were obtained by examining the records of the Kildare County Coroner. There is no central national location for the records of the 48 coroner jurisdictions in Ireland and all coroners are required by law to retain the files on each inquest indefinitely. However, the actual record of verdict given at inquest is not the one used for determining the suicide rate in the country. This is achieved by the Central Statistics Office (CSO) Form 104, which asks for the investigating police officer to give his or her opinion as to the cause of death. This results in discrepancies between what the coroner records and what the official suicide rate is presumed to be. These figures are further influenced by some coroners choosing to return a verdict "death in accordance with the medical evidence" as opposed to a verdict of suicide. The files were also examined to find the high-risk groups or those groups which have a tendency towards suicide. Over 84% of suicides were male and 32 men were between the ages of 20 and 30. It is suggested that the standardisation of recording verdicts of suicide be implemented as soon as possible as the current situation leads to variances between coroner's records and those kept by the CSO. PMID- 15522638 TI - Eruption times of permanent teeth in teenage boys and girls in Izmir (Turkey). AB - In a study in the Aegean city of Izmir, Turkey, during the summer of 2002, 2101 patients (1046 boys and 1055 girls) were examined. The examination occurred within the framework of a dental check-up performed at two schools and one pre school unit. The age span ranged from 3.98 to 24.91 years. Sex, age and present permanent teeth were recorded. Wisdom teeth were excluded. The sequence of tooth eruptions differs significantly in the lower and upper jaw, whereas no significant differences existed when comparing the sides of each jaw. The times of tooth eruption is earlier in females than in males. The entire tooth eruption process of the second dentition occurs in females earlier than in males. The tooth eruption in the lower and upper jaw of male and female probands is symmetrical. In comparing the upper and the lower jaw of both genders, it becomes evident that there is a tendency for earlier tooth eruptions in the lower jaw. In respect to the tooth eruption sequence, a change was noted in the upper jaw. Contrary to the reports of other authors, the second premolar has changed places with the canine and erupts prior to this tooth. Otherwise no major differences concerning the sequence of tooth eruptions, when compared to the results gained from other populations, were observed. Concerning the entire dentition, no acceleration of the tooth eruption could be noted. The computed differences of teeth eruption as a mean value calculated over all teeth, was +/- 1 year at maximum, compared to studies from different continents. Oral examination of teeth is a simple tool to calculate tooth eruption intervals. This first investigation on a population of Izmir revealed a change in the eruption sequence of permanent teeth. These findings are relevant for dental treatment planning and forensic odontology and should be reconfirmed at certain intervals. PMID- 15522639 TI - City centre violence. AB - Emergency medicine departments have often proved to be an important source of information concerning violence. These resulting studies of violence have not previously focused on city centre violence, but instead described violence from an entire casualty department or police district. Results from the Aarhus violence study of 1999-2000 revealed differences in characteristics of victims, place of assault and alcohol consumption when violence in the city was compared to violence elsewhere. The present study constitutes a one-year period (1999 2000), during which a total of 1106 assault victims were registered, and among these 230 were assaulted downtown. Forty-six percent of victims from the city centre were assaulted in public houses or streets nearby compared to 5% in the district outside city centre. Both in- and outside the city centre the number of victims decreased significantly in relation to the previous period. However, women who experienced violence in the city were then more likely to be assaulted in the streets, when compared to previous studies. Victims who were assaulted in the city were more frequently under the influence of alcohol than victims from the outside. It is believed that the present results are comparable with other major cities. PMID- 15522640 TI - Deaths due to terrorist bombings in Istanbul (Turkey). AB - Our study was based on a retrospective analysis of terrorist bombing related deaths in Istanbul (Turkey) between 1976 and 2000. A total of 45,714 autopsy case reports from the Morgue Department of Council of Forensic Medicine were reviewed for this study. There were 120 (0.26%) deaths due to terrorist bombing. Terrorist bombing related deaths were significantly higher during the years of increasing terrorism events in Turkey. Ninety-eight (81.7%) of cases were male and 22 (18.3%) of cases were female. The most frequent age groups of victims and terrorists was between 21 and 30 years of age (n=60; 50% of the total cases). In 49 events only one death occurred. The highest mortality was 24 in a single event. In this study, we have renewed of characteristics traumatic lesion to ascertain the cause of deaths and other autopsy findings in victims and terrorists. We also emphasize that scene investigation and medico-legal autopsy are the most important procedures provide appropriate information about deaths due to terrorist bombings. PMID- 15522641 TI - A case of suspected child sexual abuse. AB - Cases of suspected child sexual abuse are notoriously difficult in respect of investigation, management, diagnosis and aftercare. This specialised area of forensic medicine is what might be regarded as a "dynamic science" in that our understanding and knowledge of the subject is incomplete. Recent research continues apace and while this undoubtedly assists in our understanding of the complexities involved, we may be obliged to re-examine our medical opinion on the basis of emerging validated scientific research. Particularly difficult is the problem of educating non-medical professionals that the majority of children who have been the victims of child sexual abuse show no clinical evidence that abuse has occurred. These highly emotive cases may be made yet more difficult by any variation in medical opinion, not specifically limited to that between prosecution and defence "experts". Consequently it behoves us to follow clearly defined guidelines and practices in our investigation and management of these cases such that we might provide the children involved with the best care for their immediate and future health and safety. PMID- 15522642 TI - How to evaluate manual therapy: value and pitfalls of randomized clinical trials. PMID- 15522643 TI - Foot orthotics in the treatment of lower limb conditions: a musculoskeletal physiotherapy perspective. AB - Orthotic therapy is frequently advocated for the treatment of musculoskeletal pain and injury of the lower limb. The clinical efficacy, mechanical effects, and underlying mechanism of the action of foot orthotics has not been conclusively determined making it difficult for practitioners to agree on a reliable and valid clinical approach to their application and indeed even their fabrication. This problem is compounded by evidence suggesting that the most commonly used approach for orthotic prescription, the (Biomechanical Evaluation of the Foot. Vol. 1. Clinical Biomechanics Corporation, Los Angeles, 1971) approach, has poor validity and many of the associated clinical measurements of that approach lack adequate levels of reliability. This paper proposes a new approach that is based on two key elements. One is the identification, verification and quantification of physical tasks that serve as client specific outcome measures. The second is the application of specific physical manipulations during the performance of these physical tasks. The physical manipulations are selected on the basis of motion dysfunction and their immediate effects on the client specific outcome measures serve as the basis to making an informed decision on the propriety of using orthotics in individual clients. The motion dysfunction also guides the type of orthotic that is applied. Practical case examples as well as generic and specific guidelines to the application of this clinical assessment process and orthotics are provided in this paper. PMID- 15522644 TI - The flexion-rotation test and active cervical mobility--a comparative measurement study in cervicogenic headache. AB - A single blind, age and gender matched, comparative measurement study was designed to assess active range of cervical motion and passive range of rotation in cervical flexion in asymptomatic and cervicogenic headache subjects. Both procedures are commonly used in clinical practice to evaluate patients with cervicogenic headache. We studied 20 women and eight men with side dominant cervicogenic headache (mean age 43.3 years) matched with 28 asymptomatic subjects. Two experienced manipulative therapists, who were blind to each other's measurement, noted active ranges of cervical motion and passive cervical rotation performed in the flexion-rotation test using the Cervical Range of Motion Device. Headache severity was assessed by a questionnaire. Additionally, one therapist prior to neck motion assessment determined the dominant symptomatic cervical motion segment. Active cervical motion in each direction was identical between the cervicogenic and control groups. In contrast, average rotation in flexion was 44 degrees to each side in the asymptomatic group and 28 degrees towards the headache side in the symptomatic group. C1-2 was deemed to be the dominant segmental level of headache origin in 24 of 28 subjects. In those 24 subjects range of rotation during the flexion-rotation test was inversely correlated to headache severity. PMID- 15522645 TI - Anatomical relationships between selected segmental muscles of the lumbar spine in the context of multi-planar segmental motion: a preliminary investigation. AB - In the last decade, concepts regarding spinal stability have been redefined. Whereas traditional stability models considered only the integrity of the intervertebral disc and spinal ligaments, mechanisms contributing to spinal stability are now thought to include neural and muscular elements. Lumbar muscles capable of generating intersegmental stiffness are considered necessary for the control of multi-planar segmental spinal motion. The transversus abdominis, psoas, quadratus lumborum and multifidus have each been described functionally as contributing to segmental motion control in the lumbar spine. However, the fundamental anatomy of these muscles has not been fully established nor have their architectural characteristics as a functional group been explored. A dissection of the lumbar spine was undertaken to document the attachments of the deep vertebral muscles and illustrate their group architectural characteristics in the context of multi-planar segmental motion. The transversus abdominis, psoas, quadratus lumborum and multifidus were each noted to have segmental attachment patterns in the lumbar spine. As a group, they surround the lumbar motion segments from the anterolateral aspect of a vertebral body to the spinous process. A hypothetical role for this muscle group in maintaining lumbar spine stability is discussed as are suggestions for future research. PMID- 15522646 TI - Spinal kinematics and trunk muscle activity in cyclists: a comparison between healthy controls and non-specific chronic low back pain subjects-a pilot investigation. AB - The aim of this pilot study was to examine whether differences existed in spinal kinematics and trunk muscle activity in cyclists with and without non-specific chronic low back pain (NSCLBP). Cyclists are known to be vulnerable to low back pain (LBP) however, the aetiology of this problem has not been adequately researched. Causative factors are thought to be prolonged forward flexion, flexion-relaxation or overactivation of the erector spinae, mechanical creep and generation of high mechanical loads while being in a flexed and rotated position. Nine asymptomatic cyclists and nine cyclists with NSCLBP with a flexion pattern disorder primarily related to cycling were tested. Spinal kinematics were measured by an electromagnetic tracking system and EMG was recorded bilaterally from selected trunk muscles. Data were collected every five minutes until back pain occurred or general discomfort prevented further cycling. Cyclists in the pain group showed a trend towards increased lower lumbar flexion and rotation with an associated loss of co-contraction of the lower lumbar multifidus. This muscle is known to be a key stabiliser of the lumbar spine. The findings suggest altered motor control and kinematics of the lower lumbar spine are associated with the development of LBP in cyclists. PMID- 15522647 TI - Is cervical spine rotation, as used in the standard vertebrobasilar insufficiency test, associated with a measureable change in intracranial vertebral artery blood flow? AB - Cervical spine rotation is used by manual therapists as a premanipulative vertebrobasilar insufficiency (VBI) test to identify patients at risk of developing VBI post-manipulation. Investigations of the effect of rotation on vertebral artery blood flow have yielded conflicting results, the validity of the test being debated. It was the aim of this study, therefore, to investigate the effects of cervical spine rotation on vertebral artery blood flow. Transcranial Doppler sonography was used to measure intracranial vertebral artery blood flow in 30 young, healthy, female subjects, with the cervical spine in the neutral position and with sustained, end-of-range rotation. Statistically significant decreases in blood flow were demonstrated with contralateral rotation particularly, in the left (45.9+/-8.5 to 41.8+/-11.6 cm/s) and right (27.8+/-6.9 to 25.2+/-8.2 cm/s) vertebral arteries. Despite this change in blood flow, signs and symptoms of VBI were not demonstrated in these subjects. Nevertheless, these findings are of clinical importance, especially in patients who may have underlying vascular pathology. Thus, this study supports the use of the VBI test, in the absence of a more specific, sensitive and valid test, to assess the adequacy of hindbrain blood supply to identify those patients who may be at risk of serious complications post-manipulation. PMID- 15522648 TI - The patient-centredness of evidence-based practice. A case example to discuss the clinical application of the bio-psychosocial model. PMID- 15522649 TI - Let kids be kids. PMID- 15522650 TI - RNA interference, DNA methylation, and gene silencing: a bright future for cancer therapy? PMID- 15522651 TI - Cardiotoxicity after liposomal anthracyclines. PMID- 15522652 TI - Pancreatic cancer and thromboembolic disease. AB - Thromboembolic disease is a common complication of pancreatic cancer and is causally associated with the generation of an intrinsic hypercoagulable state. Pancreatic-cancer cells activate platelets and express several procoagulant factors, including tissue factor and thrombin. The activation of coagulation is not simply an epiphenomenon, but might also be related to enhanced tumour growth and angiogenesis. Clinical manifestations of thromboembolic disease in pancreatic cancer include deep venous thrombosis, pulmonary embolism, disseminated intravascular coagulation, portal vein thrombosis, and arterial thromboembolism. Reported incidences of disease range from 17% to 57%. Treatment options include warfarin and low-molecular-weight heparins. Studies over the past decade suggest that long-term use of these heparins in both primary and secondary prevention of venous thromboembolic disease improves outcomes in comparison with warfarin. Further research is needed to understand better the morbidity and mortality associated with this disease in pancreatic cancer and to optimise strategies of prevention and treatment. PMID- 15522653 TI - Cutaneous metastases in a case of fallopian-tube carcinoma. PMID- 15522654 TI - New therapeutic approaches for adult T-cell leukaemia. AB - Adult T-cell leukaemia or lymphoma is an aggressive malignant disease of mature activated T cells caused by human T-cell lymphotropic virus type I. Patients with this disease have a very poor outlook because of intrinsic chemoresistance and severe immunosuppression. In acute adult T-cell leukaemia, clinical trials in Japan show that although non-targeted combinations of chemotherapy improve response, they do not have a significant effect on complete remission and survival. Antiretroviral therapy with combination zidovudine and interferon alfa, which induces a high rate of complete remission and lengthens survival, should be the first treatment option in acute adult T-cell leukaemia. Patients with adult T cell lymphoma might benefit from initial aggressive chemotherapy followed by antiretroviral therapy. To prevent relapse in all patients allogeneic bone-marrow transplantation when feasible, or additional targeted therapy, should be mandatory. Based on current pathophysiology, we discuss promising new drugs such as arsenic trioxide, proteasome inhibitors, retinoids, and angiogenesis inhibitors, as well as cellular immunotherapy. PMID- 15522655 TI - Ultrasonography or palpation for detection of melanoma nodal invasion: a meta analysis. AB - Because treatment of distant melanoma metastases is not very effective, nodal spread should be diagnosed early so that therapeutic lymphadenectomy can be started as early as possible. Physical examination alone often does not detect nodal metastases and palpable nodes cannot be clasified unambiguously. Whether lymph-node ultrasonography-an inexpensive procedure-improves detection of nodal invasion during the initial staging and follow-up of patients with melanoma is controversial. We used meta-analysis techniques for diagnostic tests to assess the merit of ultrasonography and palpation in detection of nodal invasion in patients with melanoma. Five databases were screened until December, 2003. 12 studies, including 6642 patients and 18?610 paired palpation and ultrasound examinations, were eligible. The main limitations were variations in the definition of false negatives, and verification bias. Ultrasonography had a higher discriminatory power (odds ratio 1755; 95% CI 726-4238) than did palpation (21 [4-111]; p=0.0001). Furthermore, positive-likelihood ratios were 41.9 (95% CI 29-75) for ultrasonography and 4.55 (2-18) for palpation; negative-likelihood ratios were 0.024 (0.01-0.03) and 0.22 (0.06-0.31), respectively. Our results showed clearly that ultrasonography detects lymph-node invasion more accurately than palpation, and should therefore probably be used routinely in patients with melanoma. PMID- 15522656 TI - Part I: Vaccines for solid tumours. AB - Active specific immunotherapy holds great potential in the search for new therapeutic approaches for patients with cancer. Much preclinical and clinical evidence has shown that the immune system can be polarised against malignant cells by several vaccination strategies. Although no anticancer vaccine can be recommended outside clinical trials at present, tumour response and immunological findings in animals and humans should prompt researchers to investigate further the potential of this biotherapy. We summarise strategies for cancer vaccines so far implemented in the clinical setting, report the results of more than 200 clinical trials published over the past two decades, and discuss insights into preclinical tumour immunology that might aid the design of the next generation of cancer vaccines. PMID- 15522657 TI - Long-term treatment of metastatic renal-cell carcinoma with fluorouracil. PMID- 15522658 TI - Primary splenic histiocytoma associated with peripheral neutrophilia. PMID- 15522659 TI - Improvement of radiotherapy facilities in developing countries: a three-tier system with a teleradiotherapy network. AB - Incidence of cancer has been projected to rise worldwide by around 50% in the next 20 years, most of which will be in developing countries. To face this unprecedented global health problem, it is essential to plan effective health care strategies. Radiotherapy-a main component of the multidisciplinary therapeutic approach for cancer-involves a huge capital investment that can pose difficulties for developing countries. Limited resources should therefore be used effectively to meet the projected requirements and to find ways to offer quality assured radiotherapy to patients. Taking advantage of the global explosion of telecommunication technology, we present a proposal for creation of an integrated three-tier radiotherapy service, which consists of primary, secondary, and tertiary radiotherapy centres in developing countries-coordinated through a teleradiotherapy network. Such a network could be cost effective, help to bridge the gap, and give all patients access to the state-of-the-art technology in radiotherapy. PMID- 15522660 TI - Jobs worth doing. PMID- 15522661 TI - Molecular approaches to malaria: MAM 2004 and beyond. AB - This special issue of Trends in Parasitology comprises a collection of timely reviews arising from the 2nd Molecular Approaches to Malaria meeting held 1-5 February 2004 in Lorne, Australia, four years after the successful inaugural meeting. As the name suggests, Molecular Approaches to Malaria focused on the latest molecular developments in malaria research, and their biological and clinical implications. By no means is this special issue intended to represent a comprehensive recapitulation of all of the presentations at the meeting. Rather, the articles address, in more general terms, recent advances on broader themes that were prominent at Molecular Approaches to Malaria meeting 2004. PMID- 15522662 TI - Curation of the Plasmodium falciparum genome. AB - The malaria genome has proved invaluable to researchers worldwide in the continuing fight against malaria by stimulating and underpinning molecular approaches in gene expression studies, vaccine and drug discovery research, and by providing data to facilitate hypothesis-driven research. The combination of in silico and experimental investigations has already yielded dividends by strengthening our understanding of the many facets of the malaria parasite Plasmodium falciparum. The recently initiated curation of the genome resource is a vital investment for maintaining and enhancing the use of this genomic information in the post-genomic era. PMID- 15522663 TI - The genomics of malaria infection. AB - Malaria research is now dominated by information flowing from the genome sequencing projects and the associated transcriptome- and proteome-mapping projects. As more species are sequenced, comparative and phylogenetic comparisons are improving the quality of gene finding, and are providing various approaches to the identification of genes important to parasite biology and the pathogenesis of disease. We are still in the early days of exploiting these data in a systematic way and the sheer volume of data presents daunting challenges. This article reviews the progress in using this genomic information and discusses opportunities for other approaches. PMID- 15522664 TI - Plasmodium post-genomics: an update. AB - The concept behind the first Molecular Approaches to Malaria meeting, held 1-5 February 2000 in Lorne, Australia, was ahead of its time; to convene a meeting of malaria researchers, database developers and genomics scientists, and to discuss how genomic sciences and their relevant disciplines could be applied to solve important problems in malaria research. The success of the second Molecular Approaches to Malaria meeting, held 1-5 February 2004 in the same place, together with the influence of genomics on malaria research, is testament to the vision that the organizers had at the first meeting. This review attempts to capture some of the current efforts in the post-genomics era of malaria research and highlights the approaches discussed at the Molecular Approaches to Malaria 2004 meeting. PMID- 15522665 TI - Variant surface antigens, virulence genes and the pathogenesis of malaria. AB - The first Molecular Approaches to Malaria meeting was held 2-5 February 2000 in Lorne, Australia. Following the meeting, Brian Cooke, Mats Wahlgren and Ross Coppel predicted that research into the molecular details of the mechanisms behind sequestration of parasitized erythrocytes would "become increasingly more complicated, with further interactions, receptors, ligands and functional domains". Furthermore, they cautioned that "the challenge will be not to lose ourselves in the molecular detail, but remain focused on the role of [the var genes and other multigene families] in pathogenesis of malaria". We contemplate on these statements, following the recent second Molecular Approaches to Malaria meeting, which was held at the same venue on 2-5 February 2004. PMID- 15522666 TI - Molecular and functional aspects of parasite invasion. AB - Apicomplexan parasites have evolved an efficient mechanism to gain entry into non phagocytic cells, hence challenging their hosts by the establishment of infection in immuno-privileged tissues. Gliding motility is a prerequisite for the invasive stage of most apicomplexans, allowing them to migrate across tissues, and actively invade and egress host cells. In the late 1960s, detailed morphological studies revealed that motile apicomplexans share an elaborate architecture comprising a subpellicular cytoskeleton and apical organelles. Since 1993, the development of technologies for transient and stable transfection have provided powerful tools with which to identify gene products associated with these structures and organelles, as well as to understand their functions. In combination with access to several parasite genomes, it is now possible to compare and contrast the strategies and molecular machines that have been selectively designed by distinct life stages within a species, or by different apicomplexan species, to optimize infection. PMID- 15522667 TI - Malaria parasite transmission stages: an update. AB - The Molecular Approaches to Malaria 2004 meeting provided an opportunity to see the impressive progress in all research fields and in the four years since the previous Molecular Approaches to Malaria meeting, when much of the Plasmodium falciparum genome sequence was already available. Study of the part of the Plasmodium life cycle associated with transmission through the vector, which begins with the commitment of blood-stage forms to sexual development, has been especially fruitful. This success is a result of several reasons including: (i) the availability of the genome sequence; (ii) the availability of good animal models that allow parasite culture and facile in vivo studies of many of the life cycle stages involved in transmission; (iii) the availability of genetic manipulation technologies for the animal models of malaria, as well as P. falciparum; and (iv) the ability to study lethal gene knockouts at this stage of the life cycle. PMID- 15522668 TI - Protein trafficking in Plasmodium falciparum-infected red blood cells. AB - Plasmodium falciparum inhabits a niche within the most highly terminally differentiated cell in the human body--the mature red blood cell. Life inside this normally quiescent cell offers the parasite protection from the host's immune system, but provides little in the way of cellular infrastructure. To survive and replicate in the red blood cell, the parasite exports proteins that interact with and dramatically modify the properties of the host red blood cell. As part of this process, the parasite appears to establish a system within the red blood cell cytosol that allows the correct trafficking of parasite proteins to their final cellular destinations. In this review, we examine recent developments in our understanding of the pathways and components involved in the delivery of important parasite-encoded proteins to their final destination in the host red blood cell. These complex processes are not only fundamental to the survival of malaria parasites in vivo, but are also major determinants of the unique pathogenicity of this parasite. PMID- 15522669 TI - Of malaria, metabolism and membrane transport. AB - With the sequencing of the Plasmodium falciparum genome now complete, increasing attention is turning to the function of gene products and to cell-regulatory processes. The combination of in silico analyses with modern molecular and biophysical methods is leading to rapid advances in our understanding of the mechanisms underlying the biochemistry and physiology of the parasite and its host cell. In this brief review, we present a "snap shot" of recent work in this area, with particular emphasis on aspects relevant to the development of new antimalarial drugs. PMID- 15522670 TI - Clinical features and pathogenesis of severe malaria. AB - A major change in recent years has been the recognition that severe malaria, predominantly caused by Plasmodium falciparum, is a complex multi-system disorder presenting with a range of clinical features. It is becoming apparent that syndromes such as cerebral malaria, which were previously considered relatively clear cut, are not homogenous conditions with a single pathological correlate or pathogenic process. This creates challenges both for elucidating key mechanisms of disease and for identifying suitable targets for adjunctive therapy. The development of severe malaria probably results from a combination of parasite specific factors, such as adhesion and sequestration in the vasculature and the release of bioactive molecules, together with host inflammatory responses. These include cytokine and chemokine production and cellular infiltrates. This review summarizes progress in several areas presented at a recent meeting. PMID- 15522671 TI - Malaria vaccines: if at first you don't succeed... AB - The Roll Back Malaria campaign vowed to halve the global burden of malaria in ten years but, midway into that campaign, few new malaria control tools have been introduced, and many established methods appear to be failing with effective chemotherapy being perhaps the most problematic. It has been repeatedly argued that the discovery and implementation of a safe and effective vaccine against malaria is a major priority in the control of the disease. Indeed, many malaria control experts believe that sustainable reductions in malaria control will be nigh on impossible in the absence of such a vaccine. While most would agree that we are still some way from being able to introduce a vaccine, steady progress is being made. We review here some new approaches and developments in vaccine research that were discussed at the Molecular Approaches to Malaria conference held 1-5 February 2004 in Lorne, Australia. PMID- 15522672 TI - Blue skies or stormy weather: what lies ahead for malaria research? AB - During the past ten years, our understanding of many aspects of the biology of malaria parasites has increased dramatically. In particular, the complete genome sequences of Plasmodium falciparum and Plasmodium yoelii, the availability of transcriptome and proteome profiles, and the establishment of transfection techniques for asexual-stage malaria parasites all represent major achievements from the past decade. Now that we are truly in the post-genomic phase of biological enquiry, this article highlights some of the opportunities and challenges that lie ahead, and speculates on what we should expect to achieve in the future. PMID- 15522673 TI - Reflections on SARS. PMID- 15522674 TI - From creeping eruption to hookworm-related cutaneous larva migrans. PMID- 15522675 TI - Mucosal leishmaniasis in an Indian AIDS patient. PMID- 15522676 TI - Rheumatic fever. PMID- 15522678 TI - The aetiology, origins, and diagnosis of severe acute respiratory syndrome. AB - Severe acute respiratory syndrome (SARS) is a new infectious disease that first emerged in Guangdong province, China, in November, 2002. A novel coronavirus was later identified in patients with SARS. The detection of the virus in these patients, its absence in healthy controls or other patients with atypical pneumonia, and the reproduction of a similar disease in a relevant animal model fulfilled Koch's postulates for implicating this coronavirus as the causal agent of SARS. The full genome sequence was determined within weeks of the virus's identification. The rapid progress in the aetiology, the development of laboratory diagnostic tests, and the defining of routes of viral transmission were facilitated through a unique WHO-coordinated virtual network of laboratories, which shared information on a real-time basis through daily teleconferences. Subsequent studies have indicated that the SARS coronavirus is of animal origin, that its precursor is still present in animal populations within the region, and that live-animal markets in southern China may have provided the animal-human interphase that allowed this precursor virus to adapt to human-human transmission. These findings underscore the potential for the re emergence of SARS and the need for laboratory tests for early diagnosis. However, the low viral load in the respiratory tract makes early diagnosis of SARS a diagnostic challenge, although improvements in the sensitivity of molecular diagnostic methods continue to be made. PMID- 15522679 TI - Epidemiological and genetic analysis of severe acute respiratory syndrome. AB - The severe acute respiratory syndrome (SARS) epidemics in 2002-2003 showed how quickly a novel infectious disease can spread both within communities and internationally. We have reviewed the epidemiological and genetic analyses that have been published both during and since these epidemics, and show how quickly data were collected and analyses undertaken. Key factors that determine the speed and scale of transmission of an infectious disease were estimated using statistical and mathematical modelling approaches, and phylogenetic analyses provided insights into the origin and evolution of the SARS-associated coronavirus. The SARS literature continues to grow, and it is hoped that international collaboration in the analysis of epidemiological and contact network databases will provide further insights into the spread of this newly emergent infectious disease. PMID- 15522680 TI - Preparing to prevent severe acute respiratory syndrome and other respiratory infections. AB - Globalisation and its effect on human development has rendered an environment that is conducive for the rapid international spread of severe acute respiratory syndrome (SARS), and other new infectious diseases yet to emerge. After the unprecedented multi-country outbreak of avian influenza with human cases in the winter of 2003-2004, an influenza pandemic is a current threat. A critical review of problems and solutions encountered during the 2003-2004 SARS epidemics will serve as the basis for considering national preparedness steps that can be taken to facilitate the early detection of avian influenza, and a rapid response to an influenza pandemic should it occur. PMID- 15522681 TI - Severe acute respiratory syndrome from the trenches, at a Singapore university hospital. AB - The epidemiology and virology of severe acute respiratory syndrome (SARS) have been written about many times and several guidelines on the infection control and public health measures believed necessary to control the spread of the virus have been published. However, there have been few reports of the problems that infectious disease clinicians encounter when dealing with the protean manifestations of this pathogen. This is a qualitative account of some of the issues faced by an infectious disease physician when identifying and treating patients with SARS as well as protecting other healthcare workers and patients, including: identification of the chain of contagion, early recognition of the disease in the absence of a reliable and rapid diagnostic test, appropriate use of personal protective equipment, and the use of isolation to prevent super spreading events. Many issues need to be addressed if clinicians are to be able to manage the virus should it reappear. PMID- 15522682 TI - Collateral damage: the unforeseen effects of emergency outbreak policies. AB - The 2003 outbreak of severe acute respiratory syndrome took the province of Ontario, Canada, by surprise. A lack of planning and the decentralised nature of the health-care system meant that disruptive control measures had to be put in place to control the outbreak. Several of the control strategies were difficult to implement and resulted in considerable confusion, fear, and costs. We discuss these difficulties and offer suggestions for improving outbreak planning. PMID- 15522683 TI - Seasonality of infectious diseases and severe acute respiratory syndrome-what we don't know can hurt us. AB - The novel severe acute respiratory syndrome (SARS) coronavirus caused severe disease and heavy economic losses before apparently coming under complete control. Our understanding of the forces driving seasonal disappearance and recurrence of infectious diseases remains fragmentary, thus limiting any predictions about whether, or when, SARS will recur. It is true that most established respiratory pathogens of human beings recur in wintertime, but a new appreciation for the high burden of disease in tropical areas reinforces questions about explanations resting solely on cold air or low humidity. Seasonal variation in host physiology may also contribute. Newly emergent zoonotic diseases such as ebola or pandemic strains of influenza have recurred in unpredictable patterns. Most established coronaviruses exhibit winter seasonality, with a unique ability to establish persistent infections in a minority of infected animals. Because SARS coronavirus RNA can be detected in the stool of some individuals for at least 9 weeks, recurrence of SARS from persistently shedding human or animal reservoirs is biologically plausible. PMID- 15522684 TI - Immortality through a trypanosome. PMID- 15522685 TI - Indirect electrochemical reduction of nicotinamide coenzymes. AB - Nicotinamide coenzymes nicotinamide adenine dinucleotide (NAD(+)) and nicotinamide adenine dinucleotide phosphate (NADP(+)) were electrochemically reduced to NADH and NADPH, respectively. As direct reduction of nicotinamide coenzymes leads to inactive by-products, an indirect method using (pentamethylcyclopentadienyl-2,2'-bipyridine aqua) rhodium (III) as the mediator, was applied. A phosphate buffer solution, pH 8, with 1-10 mM NAD(P)(+) and 2.5 200 microM mediator, was pumped through a glassy carbon packed bed cathode. Virtually all the NAD(P)(+) was reduced to NAD(P)H in the cell. No sign of mediator loss due to side-reactions was detected though the mediator molecules shuttled hundreds of times between the oxidised and the reduced form. Adsorption of mediator molecules on the surface of the carbon cathode was found to be important for the reduction process. Due to strong adsorption, only minute amounts of mediator were consumed. PMID- 15522686 TI - Effects of palmitic acid and cholesterol on proton transport across black lipid membranes. AB - We studied the effect of palmitic acid (PA) and cholesterol (approximately 17 wt.%) on proton translocation across asolectin (charged) and diphytanoylphosphatidylcholine (DPhPC, neutral) black lipid membranes (BLMs). Potential difference (PD), short circuit current (SCC), and conductance (G(total)) were measured with a digital electrometer. Membranes were exposed to pH gradients (0.4-2.0 units), followed by PA addition to bath (symmetrically, 40 65 microM). The membrane conductive pathway was subdivided into an unspecific and a proton-related routes. A computer program estimated the conductances (G(un) and G(H)) of the two pathways from the measured parameters. No significant differences in proton selectivity were found between DPhPC membranes and DPhPC/cholesterol membranes. By contrast, cholesterol incorporation into asolectin increases membranes selectivity to proton. Cholesterol dramatically reduced G(un) reflecting, probably, its ability of inducing order in lipid chains. In asolectin membranes, PA increases proton selectivity, probably by acting as a proton shuttle according to the model proposed by Kamp and et al. [Biochemistry 34 (1995) 11928]. Cholesterol incorporation into asolectin membranes eliminates the PA-induced increase in proton selectivity. In DPhPC and DPhPC/cholesterol membranes, PA does not affect proton selectivity. These results are discussed in terms of the presence of cardiolipin (CL) in asolectin, cholesterol/PA interactions, and cholesterol order-inducing effects on acyl chains. PMID- 15522688 TI - The effect of polyamidoamine dendrimers on human erythrocyte membrane acetylcholinesterase activity. AB - Polyamidoamine (PAMAM) dendrimers impact on activity of acetylcholinesterase was studied. It has been shown that dendrimers induce a biphasic effect: depending on their concentrations they increase or decrease enzyme activity. It may be due to two types of interactions: direct--between dendrimers and the enzyme; indirect- via a modification of the physical state of membrane phospholipids affecting the acetylcholinesterase. PMID- 15522687 TI - A novel immunosensor based on immobilization of hepatitis B surface antibody on platinum electrode modified colloidal gold and polyvinyl butyral as matrices via electrochemical impedance spectroscopy. AB - Hepatitis B surface antibody (HBsAb) was immobilized to the surface of platinum electrode modified with colloidal gold and polyvinyl butyral (PVB) as matrices to detect hepatitis B surface antigen (HBsAg) via electrochemical impedance spectroscopy (EIS). The electrochemical measurements of cyclic voltammetry and impedance spectroscopy showed that K(4)[Fe(CN)(6)]/K(3)[Fe(CN)(6)] reactions on the platinum electrode surface were blocked due to the procedures of self assembly of HBsAb-Au-PVB. The binding of a specific HBsAb to HBsAg recognition layer could be detected by measurements of the impedance change. A new strategy was introduced for improving the sensitivity of impedance measurements via the large specific surface area and high surface free energy of Au nanoparticles and the encapsulated effect of polyvinyl butyral. The results showed that this strategy caused dramatic improvement of the detection sensitivity of HBsAg and had good linear response to detect HBsAg in the range of 20-160 ng.ml(-1) with a detection limit of 7.8 ng.ml(-1). Moreover, the studied immunosensor exhibited high sensitivity and long-term stability. PMID- 15522689 TI - Surface area involved in transdermal transport of charged species due to skin electroporation. AB - The electroporative effect on the stratum corneum (SC) is highly localized. However, the fractional area for the transport of small ions and larger ionic species differs considerably during and after high voltage (HV) application. Electroporation of SC creates new aqueous pathways, accessible for small ions, such as Cl(-) and Na(+) ions. The pores are distributed across the skin surface yielding a fractional area for current flow during electroporation of up to 0.1%. An increased permeability after high voltage application persists within a fractional area on the order of 10(-3)%. The permeabilization of SC for larger, charged molecules (M > 200 g/mol) involves Joule heating and a phase transition of the long chain sphingolipids within local transport regions (LTR). The transport area for these molecules (approximately 10(-3)%) changes only negligibly after high voltage application. PMID- 15522690 TI - An amperometric hydrogen peroxide biosensor based on immobilizing horseradish peroxidase to a nano-Au monolayer supported by sol-gel derived carbon ceramic electrode. AB - A novel strategy for fabricating horseradish peroxidase (HRP)-based H(2)O(2) sensor has been developed by combining the merits of carbon sol-gel supporting matrix and nano-scaled particulate gold (nano-Au) mediator. The thiol functional group-derived carbon ceramic electrode (CCE) was first constructed using (3 mercaptopropyl) trimethoxy silane as sol-gel monomer. Then, the stable nano-Au monolayer was obtained through covalent linkage between nano-Au and thiol group on the surface of CCE. The experimental results showed that nano-Au monolayer formed not only could steadily immobilize HRP but also efficiently retain its bioactivity. Hydrogen peroxide was detected with the aid of hydroquinone mediator to transfer electrons between the electrode and HRP. The process parameters for the fabrication of the enzyme electrode and various experimental variables such as the operating potential, mediator concentration and pH of background electrolyte were explored for optimum analytical performance of the enzyme electrode. The biosensor had a fast response of less than 8 s with linear range of 1.22 x 10(-5) to 1.10 x 10(-3)mol l(-1) and a detection limit of 6.1 x 10( 6)mol l(-1). The sensitivity of the sensor for H(2)O(2) was 0.29 A l mol(-1) cm( 2). The activation energy for enzyme reaction was calculated to be 10.1 kJ mol( 1). The enzyme electrode retained 75% of its initial activity after 5 weeks storage in phosphate buffer at pH 7. PMID- 15522691 TI - Direct determination of brucine by square wave voltammetry on 4-amino-2 mercaptopyrimidine self-assembled monolayer gold electrode. AB - 4-Amino-2-mercaptopyrimidine self-assembled monolayer (AMP SAMs/Au) was prepared on a gold electrode. The AMP SAMs/Au was characterized by using attenuated total reflection-fourier transform infrared (ATR-FTIR) and A.C. Impedance. The electrochemical behavior of brucine on AMP SAMs/Au was studied by cyclic voltammetry (CV) and square wave adsorptive stripping voltammetry (SWASV). The modified electrode showed an excellent electrocatalytic activity for the redox of brucine. The catalytic current increased linearly with the concentration of brucine in the range of 4.0 x 10(-7) to 2.0 x 10(-4) mol l(-1) by square wave voltammetry response. The detection limit was 6.0 x 10(-8) mol l(-1). PMID- 15522692 TI - Direct spectroelectrochemical titration of glutathione. AB - Previous potentiometric attempts to determine the formal potential (E'(0)) of key intracellular redox buffer glutathione resulted in contradictory values. We have developed a spectroelectrochemical method using direct reduction on metal oxide electrodes. Disulfide absorbance at 258 nm was used to titrate glutathione in the thin layer cell reversibly. At conditions close to physiological ([GSH] = 0.001 0.005 mol/l, pH = 7.34; I = 0.1 mol/l; T = 298.15 K), we have measured glutathione E(0)' = -0.22 +/- 0.02 V (NHE), corroborating the results of equilibrium measurements. PMID- 15522693 TI - Study on the enhancement of catalytic activity for hemoglobin by quinhydrone in poly(o-aminophenol) film. AB - Hemoglobin (Hb) and quinhydrone (QHQ) were incorporated in poly(o-aminophenol) [o AP, POAP] film by electropolymerization of o-aminophenol in a weak acid solution containing Hb and QHQ. The nonconducting polymer film was found to be nearly rigid by piezoelectric quartz crystal (PQC) impedance. Therefore, the thickness of the Hb-QHQ-POAP film was estimated as about 104 +/- 10 nm by quartz crystal microbalance (QCM). The QHQ mediation effects on the biomacromolecule Hb entrapped in the POAP film were investigated by using cyclic voltammetry, amperometric technique and kinetic study. Cyclic voltammograms showed that the redox peaks in the Hb-QHQ-POAP film are much more reversible than those in the Hb POAP film. The response current of the Hb-QHQ-POAP film to H(2)O(2) was almost twice than that of the Hb-POAP film. The Michaelis-Menten constant and the activation energy of Hb in the Hb-QHQ-POAP film are 7.47 mM and 13.91 kJ/mol, respectively, both are smaller than that in the Hb-POAP film. These results showed that the immobilized Hb in POAP film exhibited higher catalytic activity to H(2)O(2) due to the mediation of QHQ. PMID- 15522694 TI - Strong static magnetic field effects on yeast proliferation and distribution. AB - The present study focuses on the effects of gradient magnetic fields on the behavior of yeast, such as its proliferation and mass distribution, and evaluates the effects of magnetism on materials in the yeast culture system. Yeast, Saccharomyces cerevisiae, was incubated in a liquid medium under magnetic fields (flux density B = 14 T). When yeast in a tube was exposed to 9-14 T magnetic fields with a maximum flux density gradient of dB/dx = 94 T/m, where x is the space coordinate, the rate of yeast proliferation under the magnetic fields decreased after 16 h of incubation compared to that of the control group. The physical properties of the yeast culture system were investigated to discover the mechanism responsible for the observed deceleration in yeast proliferation under magnetic fields. Gas pressure inside the yeast culture flask was compared with and without exposure to a magnetic field. The results suggested that the gas pressure inside a flask with 6 T, 60 T/m slowly increased in comparison to the pressure inside a control tube. Due to the diamagnetism of water (medium solution) and yeast, the liquid surface distinctly inclined under gradient magnetic fields, and the hydrostatic force in suspension was strengthened by the diamagnetic forces. In addition, magnetophoresis of the yeast cells in the medium solution exhibited localization of the yeast sedimentation pattern. The roles of magnetically changed gas-transport processes, hydrostatic pressures acting on the yeast, and changes in the distribution of the yeast sedimentation, as well as the possible effects of magnetic fields on yeast respiratory systems in the observed disturbance of the proliferation are discussed. PMID- 15522695 TI - Redox kinetic measurements of glutathione at the mercury electrode by means of square-wave voltammetry. The role of copper, cadmium and zinc ions. AB - The electrode reaction of glutathione (GSH) at the hanging mercury drop electrode is studied by means of square-wave voltammetry (SWV). At potentials more positive than -0.350 V (vs. Ag/AgCl (3 mol/l KCl)) the oxidation of the mercury electrode in the presence of GSH leads to creation of a sparingly soluble mercury-GSH complex that deposits onto the electrode surface. Under cathodic potential scan, the deposited complex acts as a reducible reactant, giving raise to a well defined cathodic stripping reversible SW voltammetric response. The electrode reaction can be described by the scheme: Hg(SG)(2(s))+e(-)+2H((aq))(+) = Hg((l))+2GSH((aq)). Thus, the electrode reaction provides information on both thermodynamics and kinetics of the chemical interactions of GSH with mercury. An experimental methodology for measuring the kinetics of the electrode reactions, based on the property known as "quasireversible maximum", is developed. The standard redox rate constant is 5.09, 5.75 and 5.22 cm s(-1) in a phosphate buffer at pH 5.6, 7.0 and 8.5, respectively, with a precision of +/-10%. The high rate of the electrode reaction reflects the strong affinity of GSH towards chemical interaction with mercury. The electrode reaction is particularly sensitive to the presence of heavy metal ions such as Cu(2+), Cd(2+), and Zn(2+.) The rate of the electrode reaction decreases significantly in the presence of these ions due to simultaneous interactions of GSH with the respective ion and mercury. PMID- 15522696 TI - Electrochemical cleavage of DNA in the presence of copper-sulfosalicylic acid complex. AB - Electrochemical cleavage of DNA in the presence of copper-sulfosalicylic acid [Cu(ssal)(2)(2+)] complex was studied. The cleavage was observed in a certain potential region where redox cycling of Cu(ssal)(2)(2+)/Cu(ssal)(2)(+) took place. Cu(ssal)(2)(2+) complex mediate generation of reactive oxygen species from O(2) by the Fenton reaction, these radicals are capable of damaging DNA. The cleaved DNA fragments were separated by high-performance liquid chromatography (HPLC). The experimental results indicated that the method for electrochemical cleavage of DNA by Cu(ssal)(2)(2+) complex was simple and efficient. PMID- 15522697 TI - Deactivation of bilirubin oxidase by a product of the reaction of urate and O2. AB - The "wired" bilirubin oxidase (BOD) bioelectrocatalyst is superior to pure platinum as an electrocatalyst of the four-electron electroreduction of O(2) to water. Not only is its overpotential for O(2) reduction lower, but unlike platinum, it is not affected by organic compounds like glucose. The "wired" BOD coated carbon cathode operates for >1 week at 37 degrees C in a glucose containing physiological buffer solution. One of its key applications would be in a glucose-O(2) biofuel cell, which would operate in living tissues. The cathode is, however, short-lived in serum, losing its electrocatalytic activity in a few hours. Here we show that the damaging serum component is a product of the reaction of urate and dissolved oxygen. Exclusion of urate, by application of Nafion film on the cathode, improves the stability in serum. PMID- 15522698 TI - Neuroepithelial bodies not connected to pulmonary slowly adapting stretch receptors. AB - Neuroepithelial bodies (NEBs) are believed to be connected with one of the known types of airway receptors. The present studies determined whether NEB afferents are pulmonary slowly adapting stretch receptors (SARs). NEBs are immunoreactive with antibodies against protein gene product (PGP) 9.5 and calcitonin gene related peptide (CGRP), whereas SARs are reactive with antibody to Na(+)/K(+) ATPase. Using histochemical staining in combination with confocal microscopy, we compared the morphology of NEBs and SARs in the rat. Our results show that NEBs and SARs are different in location, size, and shape. Double staining of airway tissues for PGP (or CGRP) plus Na(+)/K(+)-ATPase shows that NEBs and SARs do not co-localize. In addition, we electrophysiologically recorded single-unit activity of SARs from the cervical vagus nerve, identified their receptive fields, dissected them into blocks, and then double-stained and examined the receptor structures. We found that the blocks contain the SAR, but not NEB structures. Thus, we conclude that NEBs are not connected to SARs. PMID- 15522699 TI - Substance P expression in TRPV1 and trkA-positive dorsal root ganglion neurons innervating the mouse lung. AB - In the present study, the co-localisation of substance P (SP) with the vanilloid receptor TRPV1 and the neurotrophin receptor tyrosine kinase trkA was analysed in airway-specific murine dorsal root ganglion (DRG) neurons. DRG neurons labelled with Fast Blue were predominantly found at the segmental levels T2-T5. Immunoreactivity for the receptor TRPV1 was localized to 12% of Fast Blue labelled DRG neurons. Double-labelling immunohistochemistry revealed that a substantial number of them also co-express SP (7.6 +/- 1.1% (mean +/- S.E.M.)), whereas neurons with immunoreactivity for TRPV1 only were found in 4.4 +/- 1.3% of the retrogradely labelled neuronal population. Further analysis of retrogradely labelled neurons showed that their majority expressed trkA (62.8 +/- 1.4%), neurofilament protein 68-kDa (64.8 +/- 1.5%) or glutamate alone (19.5 +/- 1.9%). SP was always expressed in trkA-positive neurons. Based on the extent of co-localization of SP with the receptors TRPV1 and trkA in DRG airway neurons, the present study indicates that the DRG pathway may have effects on the magnitude of neurogenic inflammation in airway diseases such as asthma. PMID- 15522700 TI - Central histamine contributes to the inspiratory off-switch mechanism via H1 receptors in mice. AB - Central histaminergic neurons are distributed in areas of the medulla and pons concerned with respiratory rhythm generation, but their effects on breathing pattern are unknown. We examined breathing pattern during hypercapnic responses in wild type (WT) and H1 receptor knockout (H1RKO) mice at 9-10 weeks of age before and after vagotomy. Minute ventilation increased with PaCO(2) increase equally in both genotypes; respiratory rate response was lower and tidal volume (V(T)) response higher in H1RKO mice than in WT mice. The V(T)-inspiratory time (T(I)) relation during hypercapnia was hyperbolic in both groups, with the curve in H1RKO mice shifted right-upward. After vagotomy, the V(T)-T(I) relation was a vertical line, which shifted right in H1RKO mice. We conclude that alterations of inspiratory off-switch and respiratory rhythm generation change breathing pattern without affecting central chemosensitivity in H1RKO. Histamine might affect breathing pattern centrally via H1 receptors. PMID- 15522701 TI - Prostaglandins are responsible for the inhibition of breathing observed with a placental extract in fetal sheep. AB - We have previously observed that the infusion of a placental extract inhibits breathing movements in fetal sheep, suggesting that a placental factor may be responsible for the inhibition of fetal breathing. Our preliminary results suggested that a small peptide or a substance bound to a peptide was likely responsible for this inhibition. Since prostaglandins are found in high concentrations in the placenta, it is possible that they may be responsible for the inhibition of breathing observed with the placental extract. We hypothesized that if prostaglandins were the active factors in the placental extract, then inhibition of the production of placental prostaglandins should eliminate the activity of the extract. We infused untreated and indomethacin/ASA-treated placental extracts into the carotid artery of eight chronically instrumented fetal sheep continuously over 3 h. The concentration of all prostaglandins measured in the untreated placental extracts were significantly higher than in the indomethacin/ASA-treated extracts. Only the infusion of the untreated placental extract induced a significant decreased in the incidence of fetal breathing. Fetal plasma prostaglandins increased significantly only with the infusion of the untreated placental extracts. These findings suggest that the inhibition of breathing observed with the placental extract is likely related to prostaglandins. PMID- 15522702 TI - Breathing pattern and hypoxic sensitivity during ageing in a new model of obesity resistant rat. AB - The Lou/C rat, an inbred strain of Wistar origin, is presented both as an obesity resistant rat and as a relevant model of successful ageing. In the present study, we examined throughout ageing the breathing pattern and the ventilatory response to hypoxia in Lou/C and age-matched Wistar rats. Ventilation was assessed by whole body plethysmography in resting condition and during a hypoxic challenge (10% O(2)) at 2, 6, 12 and 24 months of age. Lou/C rats maintained constant their resting ventilation rate throughout ageing while Wistar rats tended to decrease their ventilation with advancing age. Elderly Lou/C as well as Wistar rats increased their frequency rather than their tidal volume in response to hypoxia. As Lou/C rats did not develop obesity, ageing per se surpasses the effect of fat accumulation and prevents old rats to mobilise their tidal volume in response to hypoxia. PMID- 15522703 TI - What increases type III procollagen mRNA levels in lung tissue: stress induced by changes in force or amplitude? AB - We hypothesized that stress determined by force could induce higher type III procollagen (PCIII) mRNA expression than the stress determined by amplitude. To that end, rat lung tissue strips were oscillated for 1h under different amplitudes [1, 5 and 10% of resting length (L(B)), at 0.5 x 10(-2) N] and forces (0.25 x 10(-2), 0.5 x 10(-2) and 10(-2)N, at 5% L(B)). Resistance (R), elastance (E) and hysteresivity (eta) were analysed during sinusoidal oscillations at 1Hz. After 1h of oscillation, PCIII mRNA expression was determined by Northern-blot and semiquantitative RT-PCR. Control value of PCIII mRNA was obtained from unstressed strips. E and R increased with augmenting force and decreased with increasing amplitude, while eta remained unaltered. PCIII mRNA expression increased significantly after 1h of oscillation at 10(-2)N and 5% L(B) and remained unchanged for 6h. In conclusion, the stress induced by force but not by amplitude led to the increment in PCIII mRNA expression. PMID- 15522704 TI - HIF and VEGF relationships in response to hypoxia and sciatic nerve stimulation in rat gastrocnemius. AB - To determine if hypoxia-inducible factor-1 (HIF-1) may regulate skeletal muscle vascular endothelial growth factor (VEGF) expression in response to exercise or hypoxia, rats underwent 1h sciatic nerve electrical stimulation (ES), hypoxic exposure (H) or combined stimuli. HIF-1alpha protein levels increased six-fold with maximal (8V) ES with or without H. Similar HIF-1alpha increases occurred with sub-maximal (6V and 4V) ES plus H, but not in sub-maximal ES or H alone. VEGF mRNA and protein levels increased three-fold in sub-maximal ES or H alone, six-fold in sub-maximal ES plus H, 6.3-fold with maximal ES, and 6.5-fold after maximal ES plus H. These data suggest: (1) intracellular hypoxia during normoxic exercise may exceed that during 8% oxygen breathing at rest and is more effective in stimulating HIF-1alpha; (2) HIF-1 may be an important regulator of exercise induced VEGF transcription; and (3) breathing 8% O(2) does not alter HIF-1alpha expression in skeletal muscle, implying that exercise-generated signals contribute to the regulation of HIF-1alpha and/or VEGF. PMID- 15522705 TI - The oxidative stress in response to routine incremental cycling exercise in healthy sedentary subjects. AB - The kinetics of blood markers of the oxidative stress during and after an incremental exercise until the maximal performances is not documented in healthy sedentary subjects. We studied subjects of both sexes cycling on an ergometer until or near the V(O)(2)(max) measurement, and we measured during exercise and a 30-min recovery period the plasma concentration of thiobarbituric acid reactive substances (TBARS) which explored the production of reactive oxygen species (ROS) and two antioxidants (plasma reduced ascorbic acid (RAA) and erythrocyte reduced glutathione (GSH)). Despite we noted inter-individual differences in the instants of maximal variations of TBARS, GSH, and RAA, they were all measured within the first 20 min of the post-exercise recovery period, and at the 30th min of recovery, the three ROS blood markers tended to recover their pre-exercise levels. The maximal TBARS increase was positively correlated with V(O)(2)(max) and negatively correlated with the magnitude of RAA consumption. Our results indicate the existence of an early post-exercise oxidative stress in healthy sedentary volunteers. They also show that the ROS production is proportional to the maximal aerobic power and inversely related to the consumption of plasma antioxidants. PMID- 15522706 TI - The effect of ventilation on spectral analysis of heart rate and blood pressure variability during exercise. AB - Heart rate variability (HRV) and systolic blood pressure variability (BPV) during incremental exercise at 50, 75, and 100% of previously determined ventilatory threshold (VT) were compared to that of resting controlled breathing (CB) in 12 healthy subjects. CB was matched with exercise-associated respiratory rate, tidal volume, and end-tidal CO(2) for all stages of exercise. Power in the low frequency (LF, 0.04-0.15 Hz) and high frequency (HF, >0.15-0.4 Hz) for HRV and BPV were calculated, using time-frequency domain analysis, from beat-to-beat ECG and non-invasive radial artery blood pressure, respectively. During CB absolute and normalized power in the LF and HF of HRV and BPV were not significantly changed from baseline to maximal breathing. Conversely, during exercise HRV, LF and HF power significantly decreased from baseline to 100% VT while BPV, LF and HF power significantly increased for the same period. These findings suggest that the increases in ventilation associated with incremental exercise do not significantly affect spectral analysis of cardiovascular autonomic modulation in healthy subjects. PMID- 15522708 TI - Ion-exchange chromatography method for the purification of genomic DNA fraction from Mycobacterium bovis Bacillus Calmette-Guerin. AB - The goal of this study was to provide practical strategies for purifying genomic DNA fraction from Mycobacterium bovis Bacillus Calmette-Guerin (BCG-DNA) by ion exchange chromatography. A multistep process was developed to purify BCG-DNA. The process consisted of sonication, heating, trypsin digestion, ion-exchange chromatography, gel-filter chromatography, and lyophilization. After ion-exchange chromatography, BCG-DNA was highly purified and possessed potent biological effects. The methods described were efficient and had good reproducibility. Further, this was the first reported chromatography method to purify BCG-DNA. PMID- 15522707 TI - Upper airway calibre and impedance in patients with Steinert's myotonic dystrophy. AB - Myotonic dystrophy (MD) can be responsible for increased inspiratory muscle loading, the origin of which is debated, with some authors incriminating distal lesions and others central abnormalities. Using a recent non-invasive method based on single transient pressure-wave reflection analysis, we measured central airway calibre from the mouth to the carina and respiratory impedance in a group of adults with MD, a group of patients with sleep apnoea syndrome (SAS) but no neuromuscular disease, and a group of normal controls. All participants were awake during the measurements. We found no reduction in central airway calibre in the patients with the adult form of MD, as compared to the normal controls. These data suggest that MD may be associated with peripheral airway obstruction related to alterations in the elastic properties of the lung. PMID- 15522709 TI - Determination of a neuroprotective agent (S)-(+)-BMS-204352 in human, rat and dog plasma by enantioselective liquid chromatography-tandem mass spectrometry. AB - A sensitive liquid chromatography-electrospray ionization tandem mass spectrometry method (LC/ESI/MS/MS) for the enantioselective determination of (S) (+)-BMS-204352, a potent and specific maxi-K channel opener, in human, rat and dog plasma was developed. (S)-(+)-BMS-204352, its enantiomer (R)-(--)-BMS-204353 and the internal standard (13C-deuterated racemate of (S)-(+)-BMS-204352) were extracted from plasma using toluene. Chromatographic separation for the enantiomers was achieved on a Chiralcel OD-H analytical column with a run time of 8 min. An aqueous mobile phase modifier was added post column to enhance the mass spectrometer sensitivity. ESI mass spectra were acquired in the negative mode with selected reaction monitoring. The limit of quantitation (LLOQ) is 0.10 ng/mL for human plasma assay. Samples from a clinical study and two animal studies were processed using these procedures. Based on the in vivo data, lack of inversion of (S)-(+)-BMS-204352 to (R)-(--)-BMS-204353 was demonstrated in human, rat and dog after administration of the drug. A sensitive non-enantioselective LC/ESI/MS/MS assay has also been developed for (S)-(+)-BMS-204352 which uses a similar extraction procedure with a C18 column with a limit of quantitation at 0.05 ng/mL. Human study samples were analyzed by both methods and the correlation coefficient between both sets of data is greater than 0.99. PMID- 15522710 TI - Molecular recognition based cadmium removal from human plasma. AB - Molecularly imprinted polymers (MIPs) are easy to prepare, stable, inexpensive and capable of molecular recognition. MIPs can be considered as affinity separation media. Cadmium is a carcinogenic and mutagenic element. There is no specific treatment available for acute or chronic metal poisoning. Besides supportive therapy and hemodialysis, metal poisoning is often treated with commercially available chelating agents including EDTA and dimercaprol. However, there is histopathological evidence for increased toxicity in animals when these agents are utilized. The aim of this study is to prepare ion-imprinted polymers, which can be used for the selective removal of Cd2+ ions from Cd2+-overdosed human plasma. N-Methacryloly-(L)-cysteinemethylester (MAC) was chosen as the complexing monomer. In the first step, Cd2+ was complexed with MAC and the Cd2+ imprinted p(HEMA-MAC) beads were synthesized by suspension polymerization. After that, the template (i.e., Cd2+ ions) were removed using 0.1 M thiourea solution. The specific surface area of the Cd2+ -imprinted poly(HEMA-MAC) beads was found to be 19.4 m2/g with a size range of 63-140 microm in diameter and the swelling ratio was 78%. According to the elemental analysis results, the beads contained 42.1 micromol MAC/g polymer. The maximum adsorption capacity was 32.5 micromol Cd2+/g beads. The relative selectivity coefficients of imprinted beads for Cd2+/Pb2+ and Cd2+/Zn2+ were 7.8 and 1683 times greater than non-imprinted matrix, respectively. The Cd2+-imprinted poly(HEMA-MAC) beads could be used many times without decreasing their adsorption capacities significantly. PMID- 15522711 TI - The bioactivity of 2,5-dimethoxy-4-ethylthiophenethylamine (2C-T-2) and its detection in rat urine by capillary electrophoresis combined with an on-line sample concentration technique. AB - The bioactivity of 2,5-dimethoxy-4-ethylthiophenethylamine (2C-T-2) on nitric oxide (NO) production and the proliferation of spleen and thymus lymphocytes to mitogen stimulation in mice are reported for the first time. NO production by T and B lymphocytes in spleen and T cells in the thymus of mice decreased after the oral administration of 2C-T-2. This indicates that 2C-T-2 intake may perturb both neural and immune activity since a decrease in NO production is indicative of a weakened defense function. 2C-T-2 (the parent drug) in rat urine samples was detected by means of capillary electrophoresis/UV absorbance combined with an on line sample concentration technique. When the CZE and MEKC modes were employed, the detection limit was found to be 4.5 and 5.0 microg/mL (at a 92.1% confidence level); whereas when on-line sample concentration methods, including stacking and sweeping-micellar electrokinetic chromatography were used, the detection limits were improved to 19.2 and 9.1 ng/mL, respectively. In an analysis of some actual samples from animal experiments, three male rats were administered 20 microg/g of body weight of 2C-T-2 by intra-peritoneal injection. The first- and second-day urine fractions were collected after the administration, for use in the analysis. As a result, 2.9 microg/mL and 0.25 microg/mL of 2C-T-2, respectively, were detected after ingestion of the doses. PMID- 15522712 TI - Determination of CC-5013, an analogue of thalidomide, in human plasma by liquid chromatography-mass spectrometry. AB - A high-performance liquid chromatographic assay with MS detection has been developed for the quantitative determination of the anti-angiogenic agent CC-5013 in human plasma. Sample pretreatment involved liquid-liquid extraction with acetonitrile/1-chlorobutane (4:1, v/v) solution containing the internal standard, umbelliferone. Separation of the compounds of interest was achieved on a column packed with Waters C18 Nova-Pak material (4 microm particle size; 300 mm x 3.9 mm internal diameter) using acetonitrile, de-ionized water, and glacial acetic acid in ratios of 20:80:0.1 (v/v/v) (pH 3.5) delivered at an isocratic flow rate of 1.00 ml/min. Simultaneous MS detection was performed at m/z 260.3 (CC-5013) and m/z 163.1 (umbelliferone). The calibration curve was fit to a linear response concentration data over a range of 5-1000 ng/ml using a weighting factor of 1/x. Values for accuracy and precision, obtained from four quality controls analyzed on three different days in replicates of five, ranged from 98 to 106% and from 5.5 to 15.5%, respectively. The method was successfully applied to study the pharmacokinetics of CC-5013 in a cancer patient receiving the drug as single daily dose. PMID- 15522713 TI - Metabolism of the designer drug 4-bromo-2,5-dimethoxyphenethylamine (2C-B) in mice, after acute administration. AB - 4-Bromo-2,5-dimethoxyphenethylamine (2C-B) is a psychoactive drug of abuse often sold under the general street name "Ecstasy". Recent reports on the abuse of 2C-B and analogues denote the lack of knowledge on this drug metabolism. In the present study, we investigated the metabolic profile of 2C-B in the mouse and found unchanged 2C-B and several metabolites, which could be identified by GC/MS in the mice urine. The identification of 2C-B metabolites may give important clues for the biological and toxicological effects of this drug of abuse and provides new important data for forensic analysis on samples taken from 2C-B abusers. PMID- 15522714 TI - Sensitive determination of midazolam and 1'-hydroxymidazolam in plasma by liquid liquid extraction and column-switching liquid chromatography with ultraviolet absorbance detection and its application for measuring CYP3A activity. AB - This manuscript described a new sensitive determination of midazolam and its metabolite 1'-hydroxymidazolam by automated column-switching high-performance liquid chromatography. The test compounds were extracted from 2 ml of plasma using chloroform-hexane (30:70, v/v) and the extract was injected into a column I (TSK-PW precolumn, 10 microm, 35 mm x 4.6 mm i.d.) for clean-up and column II (C18 STR ODS-II analytical column (5 microm, 150 mm x 4.6 mm i.d.) for separation. The mobile phase for separation consisted of phosphate buffer (0.02 M, pH 4.6), perchloric acid (60%) and acetonitrile (57.9:0.1:42, v/v/v) and was delivered at a flow-rate of 0.6 ml/min. The peak was detected using a UV detector set at 254 nm. The method was validated for the concentration range 0.3-100 ng/ml, and good linearity (r > 0.998) was confirmed. Intra- and inter-day coefficient variations for midazolam and 1'-hydroxymidazolam were less than 8.5 and 6.1%, respectively, at the concentrations of 0.5, 5 and 50 ng/ml for the test compounds. Relative errors ranged from -14 to 6% and mean recoveries were 78-85%. The limit of quantification was 0.5 ng/ml for each compound. This method was sensitive enough for pharmacokinetic studies measuring CYP3A activity in human volunteers following an intravenous (1 mg) and a single-oral administration (2 mg) of a subtherapeutic midazolam dose. PMID- 15522715 TI - Liquid chromatographic determination of the plasma concentrations of cefotaxime and desacetylcefotaxime in plasma of critically ill patients. AB - A method for the simultaneous determination of cefotaxime (CTX) and desacetylcefotaxime (DES) in plasma was developed, using acetonitrile protein precipitation and high-performance liquid chromatography (HPLC) with UV-detection at 285 nm. Desacetylcefotaxime was also analysed after conversion in highly acidic medium to its lactone form (DES-lactone). The chromatographic separation was performed on a C18 Aqua column. The lower limit of quantitation was 1 microg/ml for CTX and 0.5 microg/ml for DES and DES-lactone, using 25 microl of plasma samples. The linearity of the calibration curves was satisfactory as indicated by correlation coefficients of > or =0.990. The within-day and between day precisions were <12% (n = 18) for the two products and the accuracy was between 88 and 101%. The developed HPLC method was applied for CTX and DES determination in plasma samples of critically ill patients after continuous intravenous infusion of CTX. PMID- 15522716 TI - High-sensitivity capillary electrophoresis determination of inorganic anions in serum and urine using on-line preconcentration by transient isotachophoresis. AB - Concentrations of inorganic anions, both as individual species and biotransformation products, in physiological fluids are of strong concern in clinical studies. To date, analytical methodologies have either required different analytical procedures to determine these analytes in plasma and urine, or extensive sample preparation, or unconventional and often expensive detection schemes, or both. A simple and sensitive capillary electrophoresis (CE) method with direct UV detection was developed for the simultaneous determination of iodide, bromide and nitrate in human plasma and urine, with a special focus on reliable quantification of the trace serum iodide. With the latter objective, the method incorporates a transient isotachophoresis (tITP) procedure enabling an efficient on-line preconcentration of iodide (limit of detection, 1.4 microg l( 1)) as well as other moderately mobile analytes that fall into the tITP range. The analyses of both types of biofluids were performed using an acidic electrolyte system composed of 0.25 mol l(-1) sodium chloride and 7.5 mmol l(-1) cetyltrimethylammonium chloride at pH 2.2 and 0.5 mol l(-1) 2-(N morpholino)ethanesulfonate (pH 6.0) as terminating electrolyte. Relative standard deviations (R.S.D.) below 3.0% and 9.2% were obtained for within-day and between day precision, respectively. Resolution and quantification of oxalic acid was also feasible under optimized tITP-CE conditions. Sample preparation required only ultrafiltration (serum) and dilution (urine). A number of plasma and urine samples were evaluated with this assay and the iodide, bromide and nitrate concentrations were in the expected clinical concentration ranges. PMID- 15522717 TI - Investigation of matrix effects of urine on a molecularly imprinted solid-phase extraction. AB - This study investigates matrix effects on a molecularly imprinted solid-phase extraction (MISPE) method developed for the clean-up of diphenyl phosphate (a hydrolysis product of the commonly used flame retardant and plasticizer, triphenyl phosphate) in urine samples. The influence of potentially interfering compounds that naturally occur in urine was examined with respect to extraction recovery, repeatability and selectivity. The components tested were NaCl, urea, creatinine and hippuric acid. The imprinted polymer was prepared using 2 vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as crosslinker and a structural analogue of the analyte as the template molecule. The recovery of diphenyl phosphate from water standards was over 90% using MISPE, compared to less than 25% using a non-imprinted SPE (NISPE) counterpart. The selectivity of MISPE compared to NISPE was achieved in a wash step with a basic modifier in methanol. The recovery and repeatability of the MISPE method were affected most by NaCl in the tested concentrations, while urea, creatinine and hippuric acid had no significant influence. NaCl most likely weakens the binding during the loading of the sample. This effect could be suppressed by diluting the sample with a citrate buffer at pH 4.0. PMID- 15522718 TI - Determination of polyethylene glycols of different molecular weight in the stratum corneum. AB - We developed a sensitive method for determination of polyethylene glycols (PEGs) of different molecular weight (MW) in the human stratum corneum (SC) obtained by tape stripping. The analysis is based on derivatization with pentafluoropropionic anhydride (PFPA) and gas chromatography-electron capture detection (GC-ECD). The identification and quantification of PEGs was done using individual oligomers. The method showed to be suitable for studying permeability in normal and impaired skin with respect to MW in the range of 150-600 Da. PMID- 15522719 TI - Determination of 2',3'-dideoxycytidine in maternal plasma, amniotic fluid, placental and fetal tissues by high-performance liquid chromatography. AB - 2',3'-Dideoxycytidine (DDC) is a nucleoside reverse transcriptase inhibitor that has been shown to inhibit the human immunodeficiency virus (HIV). DDC is a candidate for treatment of pregnant women to prevent prenatal transmission of HIV/AIDS to their unborn children. A quick and simple high-performance liquid chromatography (HPLC) method has been developed and validated for the determination of DDC concentrations in samples collected from a pregnant rat model (maternal plasma, amniotic fluid, placental and fetal tissues). Extraction of DDC and its internal standard 2',3'-dideoxy-3'-thiacytidine (3TC) in plasma and amniotic fluid was carried out by protein precipitation. Extraction from placental and fetal homogenates was achieved by solid phase extraction using Waters Oasis HLB solid phase extraction cartridges. Chromatographic separation was achieved on a Waters Spherisorb S3W silica column (4.6 mm x 100 mm) equipped with a Phenomenex guard column. The mobile phase used was 10% methanol in water with 22 mM formic acid. The flow rate was 0.5 ml/min, and the detection wavelength was optimized at 275 nm. Under these chromatographic conditions, DDC eluted around 12 min, and 3TC eluted around 10 min. The calibration curves for each day of validation and analysis showed good linear response through the range of 0.15-75.0 microg/ml in each of the four matrices. The relative recovery for DDC in each of the matrices ranged from 87.8% to 103.0%. Acceptable intra- and inter-day assay precision (<15% R.S.D.) and accuracy (<15% error) were observed over 0.15-75.0 microg/ml for all four matrices. PMID- 15522720 TI - Use of molecularly imprinted polymers from a mixture of tetracycline and its degradation products to produce affinity membranes for the removal of tetracycline from water. AB - The possibility of introducing multiple recognition in artificial receptors by imprinting polymers, using a mixture of tetracycline (TC) and its degradation products as templates, has been examined. The recognition ability of the resulting molecularly imprinted polymer (MIP), as evaluated by batch rebinding assay, was found to be group-specific to tetracyclines, while the single tetracycline imprinted polymer (MIP-2) prepared using TC free from degradation products as the print molecule showed considerably high selectivity for doxycycline (DC) and modest selectivity for TC and other TC derivatives, oxytetracycline (OTC) and chlortetracycline (CTC). Based on the recognition property of the multiple tetracycline imprinted polymer (MIP-1), the polymer was applied in affinity membrane extraction as a class-selective adsorption phase to remove tetracyclines residues from water. For this purpose, the ground MIP was incorporated in a plasticized poly(vinyl chloride)-membrane by casting method. Affinity separation of the obtained membrane was evaluated for the extraction of tetracycline and its analogs (CTC, OTC or DC) in aqueous solutions by a dialysis method. The membrane exhibited significantly stronger extraction ability towards tetracycline and structurally related compounds than a "blank" membrane having a non-printed polymer (NIP) as the adsorption phase. The result of these membrane extraction studies also indicates that the drug saturating at the receptor sites of MIP (deposited in membrane) faster will also be released into the receptor chamber faster. These affinity membranes were able to extract tetracyclines from water at all pHs, the highest selectivity being shown at pH 7 of the feed solution, which gives the lowest flux of the drug. Moreover, presence of salt in the feed solution increases the release of tetracycline bound in membrane. The results of the present study show that imprinting simultaneous with TC and TC degradation products formed in situ as a mixture template generates the group selectivity towards tetracyclines for the polymeric material. High affinity to a class of tetracycline of the membrane fabricated with this receptor, together with its fast and simple MIP fabrication, provides good possibilities for its application in separation processes of tetracycline antibiotics, which often contaminate the aqueous environment. PMID- 15522721 TI - Investigation of the silylation of ephedrines using N-methyl-N-trimethylsilyl trifluoroacetamide. AB - This paper describes a gas chromatography (GC)--mass spectrometry (MS) method for the simultaneous quantification of ephedrine, pseudoephedrine, cathine, norephedrine and methylephedrine in urine. The sample preparation step includes solid phase extraction and derivatisation with N-methyl-N-trimethylsilyl trifluoroacetamide (MSTFA). An evaluation of various silylation conditions compatible with screening methods in doping control analysis is presented. The method was found to be well suited for quantification of ephedrines in doping control. PMID- 15522722 TI - Liquid chromatography--electrospray ionisation mass spectrometry method for the determination of escitalopram in human plasma and its application in bioequivalence study. AB - A novel liquid chromatographic--electrospray ionisation mass spectrometric (LC- ESI-MS) method has been developed for the determination of escitalopram, an antidepressant in human plasma using paroxetine as internal standard. The method involved liquid--liquid extraction of the analyte from human plasma with a mixture of diethyl ether and dichloromethane (70:30, v/v). The chromatographic separation was achieved within 7.0 min by using 2.0 mM ammonium acetate (pH 5.0)- acetonitrile (54:46, v/v) as mobile phase and a ODS YMCAQ 150 mm x 4.6 mm analytical column; the flow-rate was 1.0 ml/min. Ion signals m/z 325.0 and 330.0 for escitalopram and internal standard, were measured in the positive mode. A detailed validation of the method was performed as per USFDA guidelines and the standard curves were found to be linear in the range of 1.0-200 ng/ml with a mean correlation coefficient more than 0.99. The absolute recovery was more than 75% for both escitalopram and internal standard. The method was simple, sensitive, precise, accurate and was successfully applied to the bioequivalence study of escitalopram in healthy, male, human subjects. PMID- 15522723 TI - A three-step purification strategy for isolation of hamster TIG2 from CHO cells: characterization of two processed endogenous forms. AB - We have recently isolated a bioactive, circulating protein of human tazarotene induced gene-2 (TIG2) as the natural ligand of the orphan receptor ChemR23. Here we describe a simplified method for the isolation of hamster TIG2 protein from Chinese hamster ovary (CHO) cell supernatant. Using a heparin-affinity column followed by two reversed phase chromatography steps resulted in the isolation of pure biologically active material. Two processed bioactive forms of Chinese hamster TIG2 were identified by Edman sequencing and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) mass fingerprint analysis, representing the amino acid residues T20 to F156, and T20 to A155 of the 163 amino acid propeptide. Comparison with the predicted aa sequence indicates a mutation or modification within the C-terminal end of the peptide. PMID- 15522724 TI - Quantitation of tigecycline, a novel glycylcycline [corrected] by liquid chromatography. AB - An ion-paired HPLC assay was developed to determine tigecycline (GAR-936) concentrations in Hank's balanced salts solution, tigecycline intra-cellular concentrations in human polymorphonuclear neutrophils (PMNs) and tigecycline concentrations in human serum. Minocycline was used as internal standard, 5% trichloroacetic acid was added to lyse PMNs and also precipitate proteins in PMNs and serum. The top aqueous layer was aspirated for HPLC assay. The chromatograms were performed with a reversed-phase C18 column with UV detector. The mobile phase consisted of acetonitrile, phosphate buffer (pH 3) and 1-octanesulfonic acid at a flow rate of 1 ml/min. Good linearity and recovery were achieved over the range of standard curves. The relative standard deviations of three quality controls for intra- and inter-day precision were less than 6.4%, and the relative errors of the intra- and inter-day accuracy were less than 7.0%. Tigecycline in Hank's buffer, PMNs and serum was stable under different test conditions. This new liquid chromatography assay is a simple, accurate and reproducible method for determining tigecycline in different matrix. PMID- 15522725 TI - Development of a Micellar electrokinetic capillary chromatography method for the determination of three drugs employed in the erectile dysfunction therapy. AB - A Micellar electrokinetic capillary chromatography method is proposed for the determination of sildenafil, vardenafil and tadalafil, which are employed in oral therapy for erectile dysfunction. Optimum conditions for the separation were investigated. A background electrolyte solution consisting of 10 mM phosphate buffer adjusted to pH 12.0, sodium dodecyl sulfate (SDS) 25 mM, hydrodynamic injection, and 25 kV as separation voltage were used. Relative standard deviations (R.S.D.s) were 1.0, 1.0, 0.4% and 2.9, 2.9, 1.9% for migration time and corrected peak area (CPA) (n = 9) for sildenafil, vardenafil and tadalafil, respectively. Detection limits obtained for the three drugs ranged from 0.19 to 0.61 mg L(-1). A linear concentration range between 1 and 20 mg L(-1) was obtained. A ruggedness test of this method was checked using the fractional factorial model of Plackett-Burman, in which the influence of six factors at three different levels was tested on different electrophoretic results: resolution and corrected peak area. The statistical evaluation of the electrophoretic results was achieved by Youden and Steiner method. The described method is rapid, sensitive and rugged and it was tested in the pharmaceutical formulations analysis obtaining recoveries between 98 and 107% respect to the nominal content. PMID- 15522726 TI - A sensitive internal standard method for the determination of melatonin in mammals using precolumn oxidation reversed-phase high-performance liquid chromatography. AB - A sensitive and accurate internal standard method to determine melatonin in mammalian tissues and physiological fluids has been described. This method includes the oxidation of melatonin to a highly fluorescent compound, N-[(6 methoxy-4-oxo-1,4-dihydroquinolin-3-yl)methyl]acetamide (6-MOQMA), and the determination of 6-MOQMA by a reversed-phase HPLC system. For the accurate and reliable determination, several melatonin analogs were designed and utilized as the internal standards, and ethyl and isopropyl analogs (having the corresponding alkyl group via the amide bond of melatonin instead of the methyl group) were found to be promising. Using these internal standards, highly accurate and sensitive determination could be accomplished using rat pineal gland samples, and the clear circadian rhythms are demonstrated. This method was also successfully applied to the determination of melatonin in a small amount (20 microL) of human saliva. PMID- 15522727 TI - Preparative high-performance liquid chromatographic separation and analysis of the Maltacine complex - a family of cyclic peptide antibiotics from Bacillus subtilis. AB - Purification of secondary metabolites from fermentation broths can be a challenging task both due to the complexity of the medium, inherently unstable molecular structures or by the action of enzymes present in the fermentation broth leading to poor isolation yield and loss of antibiotic activity. A combination of different purification techniques has usually been used to arrive at acceptable purities for characterisation of the target molecules. Due to rapid decay of antimicrobial activity a rapid preparative high-performance liquid chromatography (HPLC) method was developed that provided separation and resolution of a family of 18 closely related cyclic peptides within 110 min with minimal loss of activity. Characterisation of the peptides with LC-MS, UV/IR spectroscopy and amino acid analysis disclosed 20 different peptides with cyclic structures (lactones) with molecular weights between 1447.7 and 1519.8 Da. No peptide antibiotics with identical molecular weights have previously been reported in the literature, which lead us to conclude that this peptide complex has not been discovered before. We have named them Maltacines. PMID- 15522728 TI - Application of high-performance capillary electrophoresis to the purification process of Escherichia coli K4 polysaccharide. AB - The high-performance capillary electrophoresis (HPCE) (electrokinetic chromatography with sodium dodecyl sulphate) technique was applied to the extraction and purification process of the K4 polysaccharide from cultured bacteria in several stages. HPCE proved to be a technique with high resolution and sensitivity in analyzing K4 polysaccharide during its purification, in particular by using a strong anion-exchange resin. This is of paramount importance to monitor the product during the extraction and purification process or to test the purity of the final product. Furthermore, HPCE is able to verify that the extraction and purification process adopted is not carried out under drastic conditions capable of inducing fructose removal from the polysaccharide backbone. PMID- 15522729 TI - Development of a high performance liquid chromatography/mass spectroscopy method for the determination of strychnine concentrations in insects used to assess potential risks to insectivores. AB - A high performance liquid chromatography/mass spectrometry (HPLC/MS) method was developed in support of a study to assess potential tertiary risks posed to insectivores by strychnine baited pocket gophers (Thomomys sp.). Necropholous insects are primary consumers of pocket gopher carcasses. A field study was conducted to collect insects from strychnine-baited and control pocket gopher carcasses. The majority of the insects collected were from the orders Diptera (flies, assayed separately as adults and larvae), Coleoptera (beetles), and Hymenoptera (ants and wasps, assayed separately). Samples (0.5 g) were extracted in acetic acid (2%) and analyzed with the mass spectrometer configured for tandem mass spectrometry. For most of the samples the strychnine concentrations were less than the method limit of detection. However, strychnine concentrations as high as 0.338, 0.341, 0.698, and 0.034 microg/g were detected in ants, fly adults, fly larvae, and beetles, respectively. This information collected with the HPLC/MS method is critical for assessing potential non-target hazards for insectivores. PMID- 15522730 TI - Identification and separation of PCR products based on their GC content by denaturing high-performance liquid chromatography. AB - We show that denaturing high-performance liquid chromatography is a suitable method for the separation of DNA molecules of similar sizes but with different GC contents. A mixture of homologous molecules coming from different bacterial species may be obtained when PCR with degenerate primers is used for the amplification of a specific gene from an environmental sample. We have observed that, by selecting an appropriate temperature for the partial denaturation of the molecules, we are able to separate them according to the GC content of each DNA product. This allows us to determine if one or several types of molecules are amplified in the course of a PCR reaction. In the latter case it is possible, even with minority products, to isolate them by collecting the eluted volumes, followed by cloning, sequencing or reamplifying them by PCR, depending on the DNA concentration. We have applied this analysis to the amplification of a fragment of the ribA gene in the bacterial endosymbionts of insects, obtaining a high correlation coefficient (0.978) between retention time and the GC content of the molecules. PMID- 15522731 TI - Therapeutic applications of RNA interference: recent advances in siRNA design. PMID- 15522732 TI - The splicing of the IGF-I gene to yield different muscle growth factors. PMID- 15522733 TI - Breeding Hevea rubber: formal and molecular genetics. PMID- 15522734 TI - Gene transfer for therapeutic vascular growth in myocardial and peripheral ischemia. AB - Therapeutic vascular growth in the treatment of peripheral and myocardial ischemia has not yet fulfilled its expectations in clinical trials. Randomized, double-blinded placebo-controlled trials have predominantly shown the safety and feasibility but not the clear-cut clinically relevant efficacy of angiogenic gene or recombinant growth factor therapy. It is likely that growth factor levels achieved with single injections of recombinant protein or naked plasmid DNA are too low to induce any relevant angiogenic effects. Also, the route of administration of gene transfer vectors has not been optimal in many cases leading to low gene-transfer efficacy. Animal experiments using intramuscular or intramyocardial injections of adenovirus encoding vascular endothelial growth factor (VEGF, VEGF-A), the mature form of VEGF-D, and fibroblast growth factors (FGF-1, -2, and -4) have shown high angiogenic efficacy. Adenoviral overexpression of VEGF receptor-2 ligands, VEGF-A and the mature form of VEGF-D, enlarge the preexisting capillaries in skeletal muscle and myocardium via nitric oxide(NO)-mediated mechanisms and via proliferation of both endothelial cells and pericytes, resulting in markedly increased tissue perfusion. VEGF also enhances collateral growth, which is probably secondary to increased peripheral capillary blood flow and shear stress. As a side effect of VEGF overexpression and rapid microvessel enlargement, vascular permeability increases and may result in substantial tissue edema and pericardial effusion in the heart. Because of the transient adenoviral gene expression, the majority of angiogenic effects and side effects return to baseline by 2 weeks after the gene transfer. In contrast, VEGF overexpression lasting over 4 weeks has been shown to induce the growth of a persistent vascular network in preclinical models. To improve efficacy, the choice of the vascular growth factor, gene transfer vector, and route of administration should be optimized in future clinical trials. This review is focused on these issues. PMID- 15522735 TI - Metabolic highways of Neurospora crassa revisited. AB - This chapter describes the metabolic pathways for Neurospora crassa in the biosynthesis of amino acids, purines, pyrimidines, vitamins, and cofactors, and for glycolysis, the TCA and glyoxylate cycles and the initial stages of the pentose phosphate pathway. For each step in metabolism, the gene or genes within the genome sequence of the species is identified, correlations are made with previously identified genes, and new gene designations are assigned to others. For each gene, details given are the function of the gene product, contig location, comparison of the genetic and physical map location, Saccharomyces cerevisiae homolog, and perhaps others, and the level of similarity. PMID- 15522737 TI - Blood vessel signals during development and beyond. PMID- 15522738 TI - HIFs, hypoxia, and vascular development. AB - Cellular oxygen (O2) concentrations are tightly regulated to maintain ATP levels required for metabolic reactions in the human body. Responses to changes in O2 concentrations are primarily regulated by the transcription factor hypoxia inducible factor (HIF). HIF activates transcription of genes that increase systemic O2 delivery or provide cellular metabolic adaptation under conditions of hypoxia. HIF activity is essential for embryogenesis and various processes in postnatal life, and therefore, HIF levels need to be precisely controlled. Abnormal HIF expression is related to numerous diseases of the vascular system, including heart disease, cancer, and chronic obstructive pulmonary disease. PMID- 15522739 TI - Blood vessel patterning at the embryonic midline. AB - The reproducible pattern of blood vessels formed in vertebrate embryos has been described extensively, but only recently have we obtained the genetic and molecular tools to address the mechanisms underlying these processes. This review describes our current knowledge regarding vascular patterning around the vertebrate midline and presents data derived from frogs, zebrafish, avians, and mice. The embryonic structures implicated in midline vascular patterning, the hypochord, endoderm, notochord, and neural tube, are discussed. Moreover, several molecular signaling pathways implicated in vascular patterning, VEGF, Tie/tek, Notch, Eph/ephrin, and Semaphorin, are described. Data showing that VEGF is critical to patterning the dorsal aorta in frogs and zebrafish, and to patterning the vascular plexus that forms around the neural tube in amniotes, is presented. A more complete knowledge of vascular patterning is likely to come from the next generation of experiments using ever more sophisticated tools, and these results promise to directly impact on clinically important issues such as forming new vessels in the human body and/or in bioreactors. PMID- 15522740 TI - Wiring the vascular circuitry: from growth factors to guidance cues. PMID- 15522741 TI - Vascular endothelial growth factor and its receptors in embryonic zebrafish blood vessel development. AB - There is intense interest in how blood vessel development is regulated. A number of vascular growth factors and their receptors have been described. The vascular endothelial growth factor (VEGF) and its receptors are major contributors to normal mammalian vascular development. These receptors include VEGFR-1, VEGFR-2, VEGFR-3, neuropilin-1 (NRP1), and NRP2. The function of these genes have been determined to some degree in mouse gene targeting studies. These knockouts are embryonically lethal, and early death can be attributed in part to lack of normal blood and lymphatic vessel development. More recently, it has been demonstrated that zebrafish are an excellent model for studying the genes and proteins that regulate embryonic vascular development. Zebrafish have a number of advantages compared to mice, including rapid embryonic development and the ability to examine and manipulate embryos outside of the animal. In this review, we describe some of the earlier mouse VEGF/receptor functional studies and emphasize the development of the zebrafish vasculature. We describe the zebrafish vasculature, zebrafish VEGF and VEGF receptors, advantages of the zebrafish model, resources, and methods of determining growth factor and receptor function. PMID- 15522742 TI - Vascular extracellular matrix and aortic development. PMID- 15522743 TI - Genetics in zebrafish, mice, and humans to dissect congenital heart disease: insights in the role of VEGF. AB - Heart development and the establishment of a functional circulatory circuit are complex biological processes in which subtle perturbations may result in catastrophic consequences of cardiovascular birth defects. Studies in model organisms, most notably the mouse and the zebrafish, have identified genes that also cause these life-threatening defects when mutated in humans. Gradually, a framework for the genetic pathway controlling these events is now beginning to emerge. However, the puzzling phenotypic variability of the cardiovascular disease phenotype in humans and the recent identification of phenotypic modifiers using model organisms indicates that other genetic loci might interact to modify the disease phenotype. To illustrate this, we review the role of vascular endothelial growth factor (VEGF) during vascular and cardiac development and stress how zebrafish and mouse genetic studies have helped us to understand the role this growth factor has in human disease, in particular in the Di-George syndrome. PMID- 15522744 TI - Development of coronary vessels. PMID- 15522745 TI - Identifying early vascular genes through gene trapping in mouse embryonic stem cells. PMID- 15522746 TI - Tensile testing of a single ultrafine polymeric fiber. AB - Due to the difficulty in handling micro and nanoscale fibers and measuring the small load required for deformation, mechanical properties of these fibers have not been widely characterized. In this study, tensile test of a single-strand polycaprolactone electrospun ultrafine fiber was performed using a nano tensile tester. The tested fiber exhibited the characteristic low strength and low modulus but high extensibility at room temperature. The mechanical properties were also found to be dependent on fiber diameter. Fibers with smaller diameter had higher strength but lower ductility due to the higher 'draw ratio' that was applied during the electrospinning process. PMID- 15522747 TI - Attachment, morphology and adherence of human endothelial cells to vascular prosthesis materials under the action of shear stress. AB - In an effort to improve the long-term patency of vascular prostheses several groups now advocate seeding autologous endothelial cells (ECs) onto the lumen of the vessel prior to implantation, a procedure that involves pre-treating the prosthesis material with fibrin, collagen and/or other matrix molecules to promote cell attachment and retention. In this study, we examined the degree to which human umbilical venous endothelial cells (HUVECs) adhered to three materials commonly used polymeric vascular prosthesis that had been coated with the same commercial extra cellular matrix proteins, and after exposure to fluid shear stresses representative of femoro-distal bypass in a cone-and-plate shearing device. We quantified cell number, area of coverage and degree of cell spreading using image analysis techniques. The response of cells that adhered to the surface of each material, and following exposure to fluid shear stress, depended on surface treatment, topology and cell type. Whereas collagen coating improved primary cellular adhesion and coverage significantly, the degree of spreading depended on the underlying surface structure and on the application of the shear stress. In some cases, fewer than 30% of cells remained on the surface after only 1-h exposure to physiological levels of shear stress. The proportion of the surface that was covered by cells also decreased, despite an increase in the degree to which individual cells spread on exposure to shear stress. Moreover, the behaviour of HUVECs was distinct from that of fibroblasts, in that the human ECs were able to adapt to their environment by spreading to a much greater extent in response to shear. The quality of HUVEC attachment, as measured by extent of cell coverage and resistance to fluid shear stress, was greatest on expanded polytetrafluoroethylene samples that had been impregnated with Type I/III collagen. PMID- 15522748 TI - Regulation of mesenchymal stem cell attachment and spreading on hydroxyapatite by RGD peptides and adsorbed serum proteins. AB - The successful development of biomaterials must take into consideration how those surfaces will interact with in vivo processes such as adsorption of endogenous proteins. In this study, we examined whether modifying highly adsorbent materials like hydroxyapatite (HA) with RGD peptides would improve mesenchymal stem cell (MSC) adhesion. We found that RGD, alone, was not sufficient to promote full cell spreading. However, given that RGD-modified HA will likely adsorb osteogenic serum proteins in vivo, we evaluated MSC behavior on HA pre-coated with RGD, then over-coated with serum (RGD/FBS). Interestingly, RGD/FBS coatings additively stimulated MSC attachment and spreading compared to either coating alone, but only at low RGD coating concentrations. High RGD concentrations inhibited cell attachment, and completely eliminated cell spreading on RGD/FBS surfaces. To better understand the mechanism by which RGD and adsorbed serum proteins interactively regulate cell behavior, we monitored the deposition of fibronectin (FN) from serum onto HA pre-coated with increasing RGD concentrations. These studies showed that high RGD concentrations did not inhibit FN adsorption, therefore cell spreading is attenuated by mechanisms other than lack of FN availability. Collectively, our results suggest a potential therapeutic benefit for functionalizing HA with RGD, however such a benefit will likely depend upon the RGD density. PMID- 15522749 TI - Binding of C3 fragments on top of adsorbed plasma proteins during complement activation on a model biomaterial surface. AB - In the present study we investigate whether complement activation in blood in contact with a model biomaterial surface (polystyrene) occurs directly on the material surface or on top of an adsorbed plasma protein layer. Quartz crystal microbalance-dissipation analysis (QCM-D) complemented with enzyme immunoassays and Western blotting were used. QCM-D showed that the surface was immediately covered with a plasma protein film of approximately 8 nm. Complement activation that started concomitantly with the adsorption of the protein film was triggered by a self-limiting classical pathway activation. After adsorption of the protein film, alternative pathway activation provided the bulk of the C3b deposition that added 25% more mass to the surface. The build up of alternative pathway convertase complexes using purified C3 and factors B and D on different protein films as monitored by QCM-D showed that only adsorbed albumin, IgG, but not fibrinogen, allowed C3b binding, convertase assembly and amplification. Western blotting of eluted proteins from the material surface demonstrated that the C3 fragments were covalently bound to other proteins. This is consistent with a model in which the activation is triggered by initiating convertases formed by means of the initially adsorbed proteins and the main C3b binding is mediated by the alternative pathway on top of the adsorbed protein film. PMID- 15522750 TI - A technique for preparing protein gradients on polymeric surfaces: effects on PC12 pheochromocytoma cells. AB - A technique for preparing micropatterns and gradients of proteins on polymeric substrates has been developed in this work. Peroxides were generated on the substrate surface by UV preirradiation and they initiated graft polymerization of acrylic acid (AA) onto the surface upon a second UV irradiation. Micropatterns and gradients of poly(acrylic acid) (PAA) were formed when the substrate was placed under or moved with respect to a photomask during UV preirradiation. Protein micropatterns and gradients were fabricated on the surface by covalently linking to the carboxyl groups on PAA chains. To test cell response to the protein gradient surfaces, PC12 pheochromocytoma cells were cultured on laminin bound substrates in serum-free medium supplemented with nerve growth factor (NGF). It is found that both the attachment and neurite outgrowth behaviors of PC12 cells were dependent on the surface laminin density. However, the unreacted carboxyl groups on the polymer surface negatively affected PC12 cells. This weakened the positive influence from laminin. PMID- 15522751 TI - Craniofacial muscle engineering using a 3-dimensional phosphate glass fibre construct. AB - The current technique to replace missing craniofacial skeletal muscle is the surgical transfer of local or free flaps. This is associated with donor site morbidity, possible tissue rejection and limited supply. The alternative is to engineer autologous skeletal muscle in vitro, which can then be re-implanted into the patient. A variety of biomaterials have been used to engineer skeletal muscle with limited success. This study investigated the use of phosphate-based glass fibres as a potential scaffold material for the in vitro engineering of craniofacial skeletal muscle. Human masseter (one of the muscles of mastication)- derived cell cultures were used to seed the glass fibres, which were arranged into various configurations. Growth factors and matrix components were to used to manipulate the in vitro environment. Outcome was determined with the aid of microscopy, time-lapse footage, immunofluorescence imaging and CyQUANT proliferation, creatine kinase and protein assays. A 3-dimensional mesh arrangement of the glass fibres was the best at encouraging cell attachment and proliferation. In addition, increasing the density of the seeded cells and using Matrigel and insulin-like growth factor I enhanced the formation of prototypic muscle fibres. In conclusion, phosphate-based glass fibres can support the in vitro engineering of human craniofacial muscle. PMID- 15522752 TI - Guided cell adhesion and outgrowth in peptide-modified channels for neural tissue engineering. AB - A hydrogel scaffold of well-defined geometry was created and modified with laminin-derived peptides in an aqueous solution, thereby maintaining the geometry of the scaffold while introducing bioactive peptides that enhance cell adhesion and neurite outgrowth. By combining a fiber templating technique to create longitudinal channels with peptide modification, we were able to synthesize a scaffold that guided cell adhesion and neurite outgrowth of primary neurons. Scaffolds were designed to have numerous longitudinally oriented channels with an average channel diameter of 196 +/- 6 microm to ultimately promote fasciculation of regenerating cables and a compressive modulus of 192 +/- 8 kPa to match the modulus of the soft nerve tissue. Copolymerization of 2-hydroxylethyl methacrylate (HEMA) with 2-aminoethyl methacrylate (AEMA) scaffolds, provided primary amine groups to which two sulfhydryl terminated, laminin-derived oligopeptides, CDPGYIGSR and CQAASIKVAV, were covalently bound using the sulfo-(N maleimidomethyl)cyclohexane-1-carboxylate (sulfo-SMCC) crosslinking agent. The concentration of peptides on the scaffolds was measured at 106 +/- 4 micromol/cm(2) using the ninhydrin method and 92 +/- 9 micromol/cm(2) using the BCA protein assay. The peptide modified P(HEMA-co-AEMA) scaffolds were easily fabricated in aqueous conditions, highly reproducible, well-defined, and enhanced neural cell adhesion and guided neurite outgrowth of primary chick dorsal root ganglia neurons relative to non-peptide-modified controls. The copolymerization of AEMA with HEMA can be extended to other radically polymerized monomers and is advantageous as it facilitates scaffold modification in aqueous solutions thereby obviating the use of organic solvents which can be cytotoxic and often disrupt scaffold geometry. The combination of well-defined chemical and physical stimuli described herein provides a means for guided regeneration both in vitro and in vivo. PMID- 15522753 TI - Microencapsulation of living cells in semi-permeable membranes with covalently cross-linked layers. AB - Microencapsulation in semi-permeable membranes protects transplanted cells against immune destruction. Microcapsule strength is critical. We describe a method to microencapsulate living cells in alginate-poly-L-lysine (PLL)-alginate membranes with covalent links between adjacent layers of microcapsule membranes, while preserving the desired membrane molecular weight cut-off (MWCO) and microencapsulated cell viability. A heterobifunctional photoactivatable cross linker, N-5-azido-2-nitrobenzoyloxysuccinimide (ANB-NOS) was used. The N hydroxysuccinimide ester group of ANB-NOS was covalently linked to PLL. Islets of Langerhans were immobilized in alginate beads, incubated in PLL-ANB-NOS and again in alginate. Upon illumination with UVA, covalent links were created between the phenyl azide residue of ANB-NOS and alginate from both the core bead and the outer coating. Covalently linked microcapsules remained intact after 3 years in a strong alkaline buffer (pH 12), whereas standard microcapsules disappeared within 45 s in the same solution. A standardized mechanical stress broke 22-fold more standard than covalently linked microcapsules. The MWCO and microencapsulated cell viability were similar with standard and covalently linked microcapsules. These microcapsules, extremely resistant to chemical and mechanical stresses, will be useful in numerous applications. PMID- 15522754 TI - Fiber-based tissue-engineered scaffold for ligament replacement: design considerations and in vitro evaluation. AB - The anterior cruciate ligament (ACL) is the major intraarticular ligamentous structure of the knee, which functions as a joint stabilizer. It is the most commonly injured ligament of the knee, with over 150,000 ACL surgeries performed annually in the United States. Due to limitations associated with current grafts for ACL reconstruction, there is a significant demand for alternative graft systems. We report here the development of a biodegradable, tissue-engineered ACL graft. Several design parameters including construct architecture, porosity, degradability, and cell source were examined. This graft system is based on polymeric fibers of polylactide-co-glycolide 10:90, and it was fabricated using a novel, three-dimensional braiding technology. The resultant micro-porous scaffold exhibited optimal pore diameters (175-233 microm) for ligament tissue ingrowth, and initial mechanical properties of the construct approximate those of the native ligament. PMID- 15522755 TI - Fibroblast populated dense collagen matrices: cell migration, cell density and metalloproteinases expression. AB - Dense collagen matrices obtained by using the property of type I collagen to form liquid crystals at high concentrations, were shown to be colonized by human dermal fibroblasts (Biomaterials 23 (2002) 27). In order to evaluate them as possible tissue substitutes, we investigated in this study the mechanism of cell colonization. Fibroblasts were seeded at the surface of collagen matrices at concentrations of 5 and 40 b mg/ml. Cell density and migration were estimated from histological sections over 28 days within 500 microm thick matrices. At day 14, migration started in the 40 mg/ml matrices, attaining 320 microm in distance and 5500 cell/mm(3) in density at day 28. As zymography and western blot techniques demonstrated production of collagenase 1 (MMP1) and gelatinase A (MMP2) in culture medium, collagen hydrolysis was required for cells to penetrate the collagen network. Furthermore, the presence of MMP1 and MMP2 and their tissue inhibitors TIMP1 and TIMP2 was revealed by immunohistochemistry. We presently show that 40 mg/ml collagen matrices are colonized by human dermal fibroblasts and reach, at day 28, a density close to that measured in human dermis. PMID- 15522756 TI - The effect of the topical administration of bioactive glass on inflammatory markers of human experimental gingivitis. AB - Recent studies demonstrated that bioactive glass attenuated inflammatory reactions and bacterial growth in vitro. The aim of the present clinical study was to evaluate the effects of local bioactive glass-administration in vivo in subjects with experimental gingivitis. In each individual, contralateral teeth served as test and control over a 21-day non-hygiene (preventive phase) and a 7 day therapeutic phase. A 45S5 bioactive glass (10% solution) was applied daily (2 x 1 h) on the test teeth during the preventive and therapeutic phase of the study. Inflammation assessment was based on the plaque index record (PI), on the bleeding frequency (BOP) and the gingival crevicular fluid volume (GCF). Interleukin-1beta levels (IL-1beta) in the GCF were measured by ELISA. Bacterial deposits on teeth increased during the 21-days non-brushing period, with no difference in plaque accumulation between test and control sides. BOP levels were significantly reduced during the therapeutic phase by the application of bioactive glass. GCF-values showed a significant reduction on the test compared to control side during the preventive and therapeutic phase. IL-1beta counts decreased during the last 7 days of the study on the test side, but no significance was given. The topical application of 45S5 bioactive glass in humans with experimental gingivitis attenuated the clinical signs of inflammation, although the bacterial accumulation was not inhibited in this clinical trial. PMID- 15522757 TI - Injectability of calcium phosphate pastes. AB - A theoretical model was developed to assess ways to improve the injectability of calcium phosphate pastes. The theoretical results were then compared to experimental data obtained on calcium phosphate slips. The theoretical approach predicted that the injectability of a cement paste could be improved by an increase of the liquid-to-powder ratio, and a decrease of the particle size and the plastic limit (PL) of the powder. The theoretical results were confirmed by experimental data. Interestingly, an increase of the viscosity of the mixing liquid with small additions of xanthan had a positive effect on the paste injectability. This effect could be due to a change of the PL of the powder or to the lubricating effect of the polymer. PMID- 15522758 TI - Cytotoxicity suppression and cellular uptake enhancement of surface modified magnetic nanoparticles. AB - The aim of this study was to modify the surfaces of superparamagnetic iron oxide nanoparticles (SPION) with pullulan in order to reduce the cytotoxicity and enhance the cellular uptake of the nanoparticles. In this study, we have prepared and characterised the pullulan coated superparamagnetic iron oxide nanoparticles (Pn-SPION) of size around 40-45 nm with magnetite inner core and hydrophilic outer shell of pullulan. We have investigated the effect of cellular uptake of uncoated and Pn-SPION on cell adhesion/viability, cytotoxicity, morphology and cytoskeleton organisation of human fibroblasts. Cell cytotoxicity/adhesion studies of SPIONs on human dermal fibroblasts showed that the particles are toxic and their internalisation resulted in disruption of cytoskeleton organisation of cells. On the other hand, Pn-SPIONs were found to be non-toxic and induced changes in cytoskeleton organisation different from that observed with SPION. Transmission electron microscopy results indicated that the SPION and Pn-SPION were internalised into cells via different mechanisms, thereby suggesting that the particle endocytosis behaviour is dependent on the surface characteristics of the nanoparticles. PMID- 15522759 TI - DNA delivery from hyaluronic acid-collagen hydrogels via a substrate-mediated approach. AB - Efficient and controlled gene delivery from biodegradable materials can be employed to stimulate cellular processes that lead to tissue regeneration. In this report, a substrate-mediated approach was developed to deliver DNA from hyaluronic acid-collagen hydrogels. The hydrogels were formed by crosslinking HA with poly(ethylene glycol) diglycidyl ether. Poly(ethylene imine)(PEI)/DNA complexes were immobilized to the substrate using either biotin/neutravidin or non-specific adsorption. Complexes were formed in the presence or absence of salt to regulate complex size, and resulted in complexes with z-average diameters of 1221.7 +/- 152.3 and 139.4 +/- 1.3 nm, respectively. During 48-h incubation in PBS or hyaluronidase, DNA was released slowly from the hydrogel substrate (<30% of immobilized DNA), which was enhanced by incubation with conditioned media (approximately 50% of immobilized DNA). Transgene expression mediated by immobilized, large diameter complexes was 3 to 7-fold greater than for small diameter complexes. However, the percentage of cells expressing the transgene was greater for small diameter complexes (48.7%) than for large diameter complexes (22.3%). Spatially controlled gene transfer was achieved by topographically patterning the hydrogel to pattern cell adhesion. Biomaterial-based gene delivery can be applicable to numerous tissue engineering applications, or as a tool to examine tissue formation. PMID- 15522760 TI - Delivery of a vector encoding mouse hyaluronan synthase 2 via a crosslinked hyaluronan film. AB - We have developed a crosslinked hyaluronic acid (HA) film with DNA incorporated within its structure and have characterized this system for its efficacy in sustained transferring of a vector encoding mouse hyaluronan synthase 2 (Has2). Analysis of the DNA release kinetics indicated that the HA films degraded when treated with hyaluronidase and that they released DNA over a prolonged period of time. Gel electrophoresis revealed that this DNA was intact and immunohistochemical analysis verified the transfection capabilities of DNA release samples. The ability of released DNA encoding Has2 to promote HA synthesis was confirmed by quantifying the amount of HA produced by COS-1 cells that were transfected with release samples. The intended future application of the HA films is in prevention of post-operative peritoneal adhesions. In addition to serving as a physical barrier, the film would function as a vehicle for sustained delivery of DNA encoding Has2, which would promote the synthesis of HA in transfected tissues. PMID- 15522761 TI - Control of apatite crystal growth in a fluoride containing amelogenin-rich matrix. AB - To study how crystal growth in dental enamel is controlled by the components of the extracellular matrix, we investigated the functional roles of amelogenins and fluoride ions in apatite formation occurring through an octacalcium phosphate (OCP)-precursor pathway. Using a cation selective membrane system as a model of tooth enamel formation, we evaluated the resulting mineral habit grown in native porcine amelogenins and fluoride ions. In the absence of amelogenin and in the presence of 1 or 2 ppm F, we obtained OCP + apatite and apatite, respectively. Without amelogenins, the crystals were hexagonal prisms and cones with diameters of approximately 100-200 nm. In the presence of 10% amelogenins and in the absence of fluoride, rod-like OCP with a diameter of 35 nm were obtained. Remarkably, a combination of amelogenin and fluoride created the formation of rod like apatite crystals with dimensions similar to the former crystals. These observations indicate a cooperative role of amelogenin and fluoride in the regulation of habit, size orientation and phase of the calcium-phosphate crystals, resulting in the formation of fine rod-like apatite whose habit and orientation were similar to that of authentic tooth enamel crystals. The significant modulating effect of the amelogenin matrix combined with fluoride ions suggests the potential for this artificial system to contribute to the engineering of novel enamel-like biomaterials in vitro. PMID- 15522762 TI - Oil-in-water lipid emulsions: implications for parenteral and ocular delivering systems. AB - Lipid emulsions (LEs) are heterogenous dispersions of two immiscible liquids (oil in-water or water-in-oil) and they are subjected to various instability processes like aggregation, flocculation, coalescence and hence eventual phase separation according to the second law of thermodynamics. However, the physical stability of the LE can substantially be improved with help of suitable emulsifiers that are capable of forming a mono- or multi-layer coating film around the dispersed liquid droplets in such a way to reduce interfacial tension or to increase droplet-droplet repulsion. Depending on the concentrations of these three components (oil-water-emulsifier) and the efficiency of the emulsification equipments used to reduce droplet size, the final LE may be in the form of oil-in water (o/w), water-in-oil (w/o), micron, submicron and double or multiple emulsions (o/w/o and w/o/w). The o/w type LEs (LE) are colloidal drug carriers, which have various therapeutic applications. As an intravenous delivery system it incorporates lipophilic water non-soluble drugs, stabilize drugs that tend to undergo hydrolysis and reduce side effects of various potent drugs. When the LE is used as an ocular delivery systems they increase local bioavailability, sustain the pharmacological effect of drugs and decrease systemic side effects of the drugs. Thus, the rationale of using LE as an integral part of effective treatment is clear. Following administration of LE through these routes, the biofate of LE associated bioactive molecules are somehow related to the vehicles disposition kinetics inside blood or eyeball. However, the LE is not devoid from undergoing various bio-process while exerting their efficacious actions. The purpose of this review is therefore to give an implication of LE for parenteral and ocular delivering systems. PMID- 15522763 TI - GDSL family of serine esterases/lipases. AB - GDSL esterases and lipases are hydrolytic enzymes with multifunctional properties such as broad substrate specificity and regiospecificity. They have potential for use in the hydrolysis and synthesis of important ester compounds of pharmaceutical, food, biochemical, and biological interests. This new subclass of lipolytic enzymes possesses a distinct GDSL sequence motif different from the GxSxG motif found in many lipases. Unlike the common lipases, GDSL enzymes do not have the so called nucleophile elbow. Studies show that GDSL hydrolases have a flexible active site that appears to change conformation with the presence and binding of the different substrates, much like the induced fit mechanism proposed by Koshland. Some of the GDSL enzymes have thioesterase, protease, arylesterase, and lysophospholipase activity, yet they appear to be the same protein with similar molecular weight ( approximately 22-60 kDa for most esterases), although some have multiple glycosylation sites with higher apparent molecular weight. GDSL enzymes have five consensus sequence (I-V) and four invariant important catalytic residues Ser, Gly, Asn, and His in blocks I, II, III, and V, respectively. The oxyanion structure led to a new designation of these enzymes as SGNH-hydrolase superfamily or subfamily. Phylogenetic analysis revealed that block IIA which belonged to the SGNH-hydrolases was found only in clade I. Therefore, this family of hydrolases represents a new example of convergent evolution of lipolytic enzymes. These enzymes have little sequence homology to true lipases. Another important differentiating feature of GDSL subfamily of lipolytic enzymes is that the serine-containing motif is closer to the N-terminus unlike other lipases where the GxSxG motif is near the center. Since the first classification of these subclass or subfamily of lipases as GDSL(S) hydrolase, progress has been made in determining the consensus sequence, crystal structure, active site and oxyanion residues, secondary structure, mechanism of catalysis, and understanding the conformational changes. Nevertheless, much still needs to be done to gain better understanding of in vivo biological function, 3-D structure, how this group of enzymes evolved to utilize many different substrates, and the mechanism of reactions. Protein engineering is needed to improve the substrate specificity, enantioselectivity, specific activity, thermostability, and heterologous expression in other hosts (especially food grade microorganisms) leading to eventual large scale production and applications. We hope that this review will rekindle interest among researchers and the industry to study and find uses for these unique enzymes. PMID- 15522764 TI - Conjugated linoleic acids: are they beneficial or detrimental to health? AB - Conjugated linoleic acids (CLAs) comprise a family of positional and geometric isomers of linoleic acid (18:2n-6; LA) that are formed by biohydrogenation and oxidation processes in nature. The major dietary sources of these unusual fatty acids are foods derived from ruminant animals, in particular dairy products. The main form of CLA, cis-9, trans-11-18:2, can be produced directly by bacterial hydrogenation in the rumen or by delta-9 desaturation of the co-product vaccenic acid (trans-11-18:1) in most mammalian tissues including man. The second most abundant isomer of CLA is the trans-10, cis-12-18:2 form. Initially identified in grilled beef as a potential anti-carcinogen a surprising number of health benefits have subsequently been attributed to CLA mixtures and more recently to the main individual isoforms. It is also clear from recent studies that the two main isoforms can have different effects on metabolism and cell functions and can act through different cell signalling pathways. The majority of studies on body compositional effects (i.e. fat loss, lean gain), on cancer and cardiovascular disease attenuation, on insulin sensitivity and diabetes and on immune function have been conducted with a variety of animal models. Observations clearly emphasise that differences exist between mammalian species in their response to CLAs with mice being the most sensitive. Recent studies indicate that some but not all of the effects observed in animals also pertain to human volunteers. Reports of detrimental effects of CLA intake appear to be largely in mice and due mainly to the trans-10, cis-12 isomer. Suggestions of possible deleterious effects in man due to an increase in oxidative lipid products (isoprostanes) with trans-10, cis-12 CLA ingestion require substantiation. Unresponsiveness to antioxidants of these non-enzymatic oxidation products casts some doubt on their physiological relevance. Recent reports, albeit in the minority, that CLAs, particularly the trans-10, cis-12 isomer, can elicit pro-carcinogenic effects in animal models of colon and prostate cancer and can increase prostaglandin production in cells also warrant further investigation and critical evaluation in relation to the many published anti-cancer and anti-prostaglandin effects of CLAs. PMID- 15522765 TI - The effect of 30% oxygen on visuospatial performance and brain activation: an fMRI study. AB - This study aimed to investigate the hypothesis that administration of the air with 30% oxygen compared with normal air (21% oxygen) enhances cognitive functioning through increased activation in the brain. A visuospatial task was presented while brain images were scanned by a 3 T fMRI system. The results showed that there was an improvement in performance and also increased activation in several brain areas in the higher oxygen condition. These results suggest that a higher concentration of breathed oxygen increases saturation of blood oxygen in the brain and facilitates performance. PMID- 15522766 TI - Working memory, inhibition, and fluid intelligence as predictors of performance on Tower of Hanoi and London tasks. AB - The contributions of working memory, inhibition, and fluid intelligence to performance on the Tower of Hanoi (TOH) and Tower of London (TOL) were examined in 85 undergraduate participants. All three factors accounted for significant variance on the TOH, but only fluid intelligence accounted for significant variance on the TOL. When the contribution of fluid intelligence was accounted for, working memory and inhibition continued to account for significant variance on the TOH. These findings support argument that fluid intelligence contributes to executive functioning, but also show that the executive processes elicited by tasks vary according to task structure. PMID- 15522767 TI - Differences in the transmission of sensory input into motor output between introverts and extraverts: Behavioral and psychophysiological analyses. AB - The present study was designed to investigate extraversion-related individual differences in the speed of transmission of sensory input into motor output. In a sample of 16 introverted and 16 extraverted female volunteers, event-related potentials, lateralized readiness potentials (LRPs), and electromyogram (EMG) were recorded as participants performed a visual choice reaction time task. As additional behavioral indicators of performance, measures of reaction time (RT) and response dynamics were obtained. Although extraversion-related differences were found neither for behavioral measures nor for the N1 and P3 components of the evoked potential, introverts showed a reliably shorter latency in stimulus locked LRP than extraverts. This latter finding supports the notion of faster stimulus analysis in introverts compared to extraverts. Furthermore, there was no indication of extraversion-related individual differences in speed of response organization and response execution as indicated by response-locked LRP and EMG latencies, respectively. However, a significantly higher EMG amplitude observed with introverts pointed to a less accurately adjusted motor output system of introverts compared to extraverts. PMID- 15522768 TI - The long-term effects of mild head injury on short-term memory for visual form, spatial location, and their conjunction in well-functioning university students. AB - Research has suggested the presence of subtle long-term cognitive changes in otherwise well-functioning individuals who have previously sustained a mild head injury (MHI). This paper investigated the long-term effects of MHI on visual, spatial, and visual-spatial short-term memory in well-functioning university students. Sixteen students who reported having sustained a MHI were compared to 16 controls on tests of short-term memory (STM) for abstract polygons in haphazardly arranged locations. The three tests differed only in the requirements for recall (shapes for the visual task, locations for the spatial task, and the shapes in their respective locations for the visual-spatial task). MHI participants were selectively impaired on spatial memory, suggesting that tasks of spatial STM may be more sensitive, compared to tasks of visual STM, to the subtle long-term cognitive changes that may be present after a MHI. PMID- 15522769 TI - Increased interhemispheric interaction is associated with decreased false memories in a verbal converging semantic associates paradigm. AB - Recent evidence indicates that task and subject variables that are associated with increased interaction between the left and right cerebral hemispheres result in enhanced performance on tests of episodic memory. The current study looked at the effects of increased interhemispheric interaction on false memories using a verbal converging semantic associates paradigm. In Experiment 1, strong right handedness (which is associated with decreased interhemispheric interaction) was associated with higher rates of false memories. In Experiment 2, bilateral saccadic eye movements (which are associated with increases in interhemispheric interaction) were associated with fewer false memories. The results provide further support for an interhemispheric basis for episodic/explicit memory. PMID- 15522770 TI - Inconsistency in serial choice decision and motor reaction times dissociate in younger and older adults. AB - Intraindividual variability (inconsistency) in reaction time (RT) latencies was investigated in a group of younger (M=25.46 years) and older (M=69.29 years) men. Both groups performed 300 trials in 2-, 4-, and 8-choice RT conditions where RTs for decision and motor components of the task were recorded separately. A dissociation was evident in that inconsistency was greater in older adults for decision RTs when task demands relating to the number of choices and fatigue arising from time-on-task were high. For younger persons, a weak trend toward greater inconsistency in motor RTs was evident. The results are consistent with accounts suggesting that inconsistency in neurobiological mechanisms increases with age, and that attentional lapses or fluctuations in executive control contribute to RT inconsistency. PMID- 15522771 TI - Endogenous task shift processes in relapsing-remitting multiple sclerosis. AB - This paper reports a study that was aimed to evaluate executive functions in relapsing-remitting multiple sclerosis patients. The groups tested comprised 22 relapsing-remitting multiple sclerosis patients, and 22 non-brain damaged controls. When one is engaged in two speeded tasks, not simultaneously but with some form of alternation, it is slower to respond to an item of task A if it was preceded by an item of task B, than when it was preceded by an item of task A. Shifts between sets of cognitive operations can be internally or externally generated. Endogenous task shift refers to advance preparation for the new task. In the present study, we tested endogenous shift cost in relapsing-remitting multiple sclerosis patients. The results indicate a greater shift cost for patients than for non-brain damaged controls. PMID- 15522772 TI - Sex differences in spatial ability: a lateralization of function approach. AB - The current study was designed to examine whether the extent of the male advantage in performance on a spatial task was determined by the extent to which the task was right-hemisphere dependent. Participants included 108 right-handed men and women who completed the mental rotation, waterlevel, and paperfolding tasks, all of which were presented bilaterally. The results partially supported the hypothesis. On the mental rotation task, men showed a right-hemisphere advantage, whereas women showed no hemispheric differences; however, no overall sex differences were observed. On the waterlevel task, men outperformed women, and both men and women showed a right-hemisphere advantage. On the paperfolding task, no sex or hemispheric differences were observed. Although the findings of the current study were mixed, the study provides a framework for examining sex differences across different types of spatial ability. PMID- 15522773 TI - Size of the neocerebellar vermis is associated with cognition in healthy elderly men. AB - Cerebellar volumes show small positive correlations with cognitive ability in young adults, but no studies have examined this relationship in older adults. Furthermore, no studies have examined relationships between sizes of subareas of the cerebellum and cognitive ability. We hypothesized that size of the two areas of the neocerebellar vermis would correlate with a battery of eight cognitive tests in 50 men aged 65-70. Size of the neocerebellar areas of the vermis correlated positively with several cognitive tests (r's=.29-.37, p<.05), whilst sizes of other parts of the vermis did not correlate with any cognitive tests. Total cerebellar volumes correlated significantly with a test of nonverbal reasoning (r=.030, p=.42). These findings suggest a specific association of neocerebellar vermis size with variations in cognitive ability in older adults. PMID- 15522774 TI - Nucleotide-dependent domain motions within rings of the RecA/AAA(+) superfamily. AB - The oligomeric rings formed by RecA-fold proteins are mechanochemical motors that perform many important biological functions. Their RecA-fold domains convert the chemical energy of ATP into mechanical work through large nucleotide-dependent conformational changes. This review summarizes recent structural and mechanistic works on the F1-ATPase and HslU regarding to the force generation by individual RecA folds in the context of ring structures. The F1-ATPase ring for example generates the force perpendicular to the ring axis, while the HslU ring generates forces presumably parallel to it. There exists a strong correlation between the directions of forces generated and the orientation of the RecA folds, not only in these two proteins but also in T7 DNA helicase, suggesting that it should be possible to predict the direction of forces generated by other members of this family on the basis of the orientation of their RecA folds. PMID- 15522775 TI - Assembly of collagen into microribbons: effects of pH and electrolytes. AB - Collagen represents the major structural protein of the extracellular matrix. Elucidating the mechanism of its assembly is important for understanding many cell biological and medical processes as well as for tissue engineering and biotechnological approaches. In this work, conditions for the self-assembly of collagen type I molecules on a supporting surface were characterized. By applying hydrodynamic flow, collagen assembled into ultrathin ( approximately 3 nm) highly anisotropic ribbon-like structures coating the entire support. We call these novel collagen structures microribbons. High-resolution atomic force microscopy topographs show that subunits of these microribbons are built by fibrillar structures. The smallest units of these fibrillar structures have cross-sections of approximately 3 x 5nm, consistent with current models of collagen microfibril formation. By varying the pH and electrolyte of the buffer solution during the self-assembly process, the microfibril density and contacts formed within this network could be controlled. Under certain electrolyte compositions the microribbons and microfibers display the characteristic D-periodicity of approximately 65 nm observed for much thicker collagen fibrils. In addition to providing insight into the mechanism of collagen assembly, the ultraflat collagen matrices may also offer novel ways to bio-functionalize surfaces. PMID- 15522776 TI - Dynamics of DNA replication: an ultrastructural study. AB - DNA replication in cells takes place in domains scattered throughout the nucleoplasm. We have characterized the dynamics of DNA synthesis in synchronized mid-S-phase HeLa cells. Saponin-permeabilized cells were allowed to elongate nascent DNA chains in presence of biotin-dUTP for 5, 15, and 30 min (a pulse experiment), or for 5 min followed by an incubation with unlabeled precursors for 10 or 25 min (a pulse-and-chase experiment). The replication foci were then identified in ultrathin sections using immunogold labeling of the incorporated biotin. Total number of particles per nucleus, total scanned area of the nucleus, size, shape, and gold particle number of each labeled cluster, and the density of clusters per nucleus were evaluated. We have demonstrated that as replication proceeds, the labeled sites increase in size up to 240 nm (30 min incorporation) while maintaining a broadly round shape. In pulse-and-chase experiments the labeled DNA was shown to spread to occupy DNA foci of approximately 400 nm in diameter. These results demonstrate that DNA replication is compartmentalized within cell nuclei at the level of DNA foci and support the view that the synthetic centers are spatially constrained while the chromatin loops are dynamic during DNA synthesis. PMID- 15522777 TI - The crystal structure of Ym1 at 1.31 A resolution. AB - Upon nematode infection, murine peritoneal macrophages synthesize and secrete large amounts of the Ym1 protein, which is a unique functional marker for alternatively activated macrophages in T(H)2-mediated inflammatory responses. Ym1 shares significant structural similarity to the family 18 chitinases. Previously, Ym1 has been studied with respect to its carbohydrate-binding ability and glycosyl hydrolysis activity and this has led to various inconclusive interpretations. Our present co-crystallization and soaking experiments with various glucosamine or N-acetylglucosamine oligomers yield only the uncomplexed Ym1. The refined Ym1 structure at 1.31A resolution clearly displays a water cluster forming an extensive hydrogen bond network with the "active-site" residues. This water cluster contributes notable electron density to lower resolution maps and this might have misled and given rise to a previous proposal for a monoglucosamine-binding site for Ym1. A structural comparison of family 18 glycosidase (-like) proteins reveals a lack of several conserved residues in Ym1, and illustrates the versatility of the divergent active sites. Therefore, Ym1 may lack N-acetylglucosamine-binding affinity, and this suggests that a new direction should be taken to unravel the function of Ym1. PMID- 15522778 TI - Automated most-probable loss tomography of thick selectively stained biological specimens with quantitative measurement of resolution improvement. AB - We describe the technique and application of energy filtering, automated most probable loss (MPL) tomography to intermediate voltage electron microscopy (IVEM). We show that for thick, selectively stained biological specimens, this method produces a dramatic increase in resolution of the projections and the computed volumes versus standard unfiltered transmission electron microscopy (TEM) methods. This improvement in resolution is attributed to the reduction of chromatic aberration, which results from the large percentage of inelastic electron-scattering events for thick specimens. These improvements are particularly evident at the large tilt angles required to improve tomographic resolution in the z-direction. This method effectively increases the usable thickness of selectively stained samples that can be imaged at a given accelerating voltage by dramatically improving resolution versus unfiltered TEM and increasing signal-to-noise versus zero-loss imaging, thereby expanding the utility of the IVEM to deliver information from within specimens up to 3 microm thick. PMID- 15522779 TI - Structural and physiological effects of calcium and magnesium in Emiliania huxleyi (Lohmann) Hay and Mohler. AB - In organisms which perform both photosynthesis and calcification, the fact that calcification proceeds faster in the light than in the dark has led to the long established view that photosynthesis and calcification are closely coupled. It is now clear that calcification does not promote photosynthesis, but an enhancement of calcification by photosynthesis could still explain why calcification is faster in the light. To test this, the kinetics of the two processes were monitored over a wide range of calcium concentrations (0-50 mM) in the coccolithophore Emiliania huxleyi. The addition of 50 mM calcium strongly inhibited both processes, but when incubated in lower concentrations, rates of calcification increased up to 20 mM calcium whilst those of photosynthesis remained constant over the same range of calcium concentrations. So, rates of calcification are able to rise without a concomitant increase in photosynthetic rates. In addition, calcification rate and coccolith morphology responded similarly to changes in calcium concentrations; low calcification rates were associated with poor coccolith structure (undercalcification) and high calcification rates with perfectly formed coccoliths. Calcium concentration thus strongly influences calcification affecting both crystal structure and rate of calcite deposition. A similar structural analysis of coccoliths from cells grown in different magnesium concentrations showed that this ion is also essential for calcification, since strong signs of coccolith malformation and undercalcification were apparent at both low and high magnesium concentrations. In contrast with the calcium results, coccoliths were flawless only in the normal seawater concentration of 58 mM magnesium. We conclude that photosynthesis and calcification are not closely coupled and that calcification depends on a precise balance of both calcium and magnesium concentrations. PMID- 15522780 TI - Membrane-induced structure of the mammalian tachykinin neuropeptide gamma. AB - Neuropeptide gamma (NPgamma) is a neurokinin-2 (NK-2) receptor selective agonist, which plays an important role in mediation of asthma and elicits a wide range of biological responses like bronchoconstriction, vasodepression and regulation of endocrine functions. The structure determination of this peptide agonist is important in understanding the molecular basis of peptide ligand recognition by the receptor and for rational drug design. In the present study we report the solution structure of NPgamma characterized by circular dichroism (CD) spectropolarimetry and 2D (1)H NMR spectroscopy in both aqueous and membrane mimetic solvents. Effect of calcium ions on the conformation of NPgamma was also studied using CD spectropolarimetry. Sequence-specific resonance assignments of protons have been made with the aid of correlation spectroscopy experiments and nuclear Overhauser effect spectroscopy experiments. The distance constraints obtained from the NMR data have been utilized to generate a family of structures, which have been refined using restrained energy minimization and dynamics. These data show that in water NPgamma prefers to be in an extended chain conformation whereas a helical conformation is induced in the central core and the C-terminal region of the peptide (K13-M21) in the presence of perdeuterated dodecylphosphocholine micelles, a membrane model system. A type II' beta turn from H9 to R11 precedes the helical core in the C-terminus of NPgamma. N-terminus of NPgamma also displays some degree of order and a possible turn structure. Conformation adopted by NPgamma in presence of lipid micelles represents a structural motif typical of NK-2 selective agonists and is similar to that observed for Neurokinin A in hydrophobic environment. The observed conformational features have been correlated to the binding ability and biological activity of NPgamma. PMID- 15522781 TI - Position of single amino acid substitutions in the collagen triple helix determines their effect on structure of collagen fibrils. AB - Collagen II fibrils are a critical structural component of the extracellular matrix of cartilage providing the tissue with its unique biomechanical properties. The self-assembly of collagen molecules into fibrils is a spontaneous process that depends on site-specific binding between specific domains belonging to interacting molecules. These interactions can be altered by mutations in the COL2A1 gene found in patients with a variety of heritable cartilage disorders known as chondrodysplasias. Employing recombinant procollagen II, we studied the effects of R75C or R789C mutations on fibril formation. We determined that both R75C and R789C mutants were incorporated into collagen assemblies. The effects of the R75C and R789C substitutions on fibril formation differed significantly. The R75C substitution located in the thermolabile region of collagen II had no major effect on the fibril formation process or the morphology of fibrils. In contrast, the R789C substitution located in the thermostable region of collagen II caused profound changes in the morphology of collagen assemblies. These results provide a basis for identifying pathways leading from single amino acid substitutions in collagen II to changes in the structure of individual fibrils and in the organization of collagenous matrices. PMID- 15522782 TI - Crystallography and mutagenesis point to an essential role for the N-terminus of human mitochondrial ClpP. AB - We have determined a 2.1 A crystal structure for human mitochondrial ClpP (hClpP), the proteolytic component of the ATP-dependent ClpXP protease. HClpP has a structure similar to that of the bacterial enzyme, with the proteolytic active sites sequestered within an aqueous chamber formed by face-to-face assembly of the two heptameric rings. The hydrophobic N-terminal peptides of the subunits are bound within the narrow (12 A) axial channel, positioned to interact with unfolded substrates translocated there by the associated ClpX chaperone. Mutation or deletion of these residues causes a drastic decrease in ClpX-mediated protein and peptide degradation. Residues 8-16 form a mobile loop that extends above the ring surface and is also required for activity. The 28 amino acid C-terminal domain, a unique feature of mammalian ClpP proteins, lies on the periphery of the ring, with its proximal portion forming a loop that extends out from the ring surface. Residues at the start of the C-terminal domain impinge on subunit interfaces within the ring and affect heptamer assembly and stability. We propose that the N-terminal peptide of ClpP is a structural component of the substrate translocation channel and may play an important functional role as well. PMID- 15522783 TI - Electron paramagnetic resonance spectroscopy and molecular modelling of the interaction of myelin basic protein (MBP) with calmodulin (CaM)-diversity and conformational adaptability of MBP CaM-targets. AB - The classic 18.5 kDa isoform of murine myelin basic protein (mMBP) has been shown to bind calmodulin (CaM) strongly and specifically in vitro. Here, we have used site-directed spin labelling (SDSL) and electron paramagnetic resonance (EPR) spectroscopy to map more precisely the sites of interaction of recombinant mMBP (rmMBP) with CaM. On the basis of these and previous experimental data, and the predictions of CaM-binding motifs using the Calmodulin Target Database (), three main segments of MBP were suggested for the interaction. The first site is located at the C-terminus; the second one lies in the central portion of the protein and forms an amphipathic alpha-helix in reconstituted myelin-mimetic systems; the third is quite close to the N-terminus. The murine Golli-MBP isoform J37 has also been shown to bind CaM in vitro, and an interaction site was predicted in the N-terminal Golli-specific portion of the protein. From these four segments, we selected peptide fragments of 12-14 residues in length, chosen on the bases of their amphipathicity and CaM-target characteristics. We modelled each of these peptides as alpha-helices, and performed docking simulations to investigate their interactions with the CaM peptide-binding tunnel. Different yet almost equally favourable CaM-binding modes were found for each of them. The experimental SDSL/EPR and theoretical modelling results were in good agreement, and supported the conjecture that there are several plausible CaM-binding sites in MBP, that could be induced into an alpha-helical conformation by their interaction with CaM and account for strong immobilisation of spin-labeled residues in all three segments. Phosphorylation and deimination were also emulated and simulated for known sites of MBP post-translational modification. The results obtained confirmed the appropriate utilisation of simple residue substitutions to mimic the natural modifications, and demonstrated molecular mechanisms by which MBP-CaM interactions could be modulated in vivo. PMID- 15522784 TI - Crystal structure of KD93, a novel protein expressed in human hematopoietic stem/progenitor cells. AB - The crystal structure of a novel hypothetical protein, KD93, expressed in human hematopoietic stem/progenitor cells, was determined at 1.9A resolution using the multiple-wavelength anomalous dispersion (MAD) method. The protein KD93, which is encoded by the open reading frame HSPC031, is a NIP7 homologue and belongs to the UPF0113 family. The structural and functional information for the group of homologues has not yet been determined. Crystallographic analysis revealed that the overall fold of KD93 consists of two interlinked alpha/beta domains. Structure-based homology analysis with DALI revealed that the C domain of KD93 matches the PUA domain of some RNA modification enzymes, especially that of archaeosine tRNA-ribosyltransferase (ArcTGT), which suggests that its possible molecular function is related to RNA binding. The difference between the RNA binding regions of KD93 and ArcTGT in amino acid constitution and surface electrostatic potential indicate that they may have different RNA binding modes. The N domain of KD93 is a unique structure with no obvious similarity to other proteins with known three-dimensional structures. The high-resolution structure of KD93 provides a first view of a member of the family of hypothetical proteins. And the structure provides a framework to deduce and assay the molecular function of other proteins of the UPF0113 family. PMID- 15522785 TI - Phylogeography of the false smooth snakes, Macroprotodon (Serpentes, Colubridae): mitochondrial DNA sequences show European populations arrived recently from Northwest Africa. AB - Mitochondrial DNA (1075 bp: cytochrome b, 300 bp; 12S rRNA, 393 bp; and 16S rRNA, 382 bp) corroborates the monophyly of the genus Macroprotodon and of the species M. mauritanicus, M. abubakeri, and M. brevis. The subspecies M. brevis ibericus is also monophyletic. The mtDNA tree presented here indicates that M. cucullatus consists of at least two separate units and may possibly represent a primitive morphology rather than a species in its own right. However, this hypothesis is tentative since it is only reflects the history of a single evolutionary unit (mtDNA). A definitive understanding of the evolution of M. cucullatus will not be possible until informative nuclear markers are added to the mitochondrial data. Macroprotodon appears to have originated in the Maghreb region of NW Africa and speciated there around 4-5.5 million years ago around the end of the Miocene period, after which its three main lineages may each have expanded north into more mesic conditions. The group also spread eastwards into coastal areas of Libya quite recently and on to Egypt and Israel. Later still, M. b. ibericus from extreme north Morocco reached the Iberian Peninsula, and M. mauritanicus from Tunisia or Algeria colonised the Balearic Islands of Menorca and Mallorca. Both these range extensions may result from very recent natural colonisations or even from accidental human introduction. Recency of origin of Iberian and Balearic populations is indicated by uniformity of their mtDNA even across large distances, and its great similarity to that of populations in source regions. Isolated populations assigned to M. cucullatus in the Hoggar mountains (southern Algeria) and Western Sahara are probably relicts from quite recent periods of climatic amelioration in the North African desert. PMID- 15522786 TI - Widespread polyphyly among Alopiinae snail genera: when phylogeny mirrors biogeography more closely than morphology. AB - Consider a group of species that is evenly divided by an easily identifiable complex morphological character. Most biologists would assume that this character should provide better phylogenetic information than, say, the spatial distribution of these species over a fairly continuous 500-km radius area. Paradoxically, this is not the case among terrestrial snail genera in the clausiliid subfamily Alopiinae. Phylogenetic analysis using the nuclear markers ITS1/ITS2 and mitochondrial markers COI/12S reveals widespread homoplasy in the clausilial apparatus (a complex aperture-closing mechanism), and concomitant extensive polyphyly among Carinigera, Isabellaria, and Sericata. In contrast, phylogenetic relationships as revealed by molecular data are closely congruent with biogeography at a relatively small scale. A combination of extremely low vagility and extremely high morphological convergence has conspired to produce this unexpected result. Implications as to the function of the clausilial apparatus are discussed. PMID- 15522787 TI - Molecular phylogeography and systematics of the arid-zone members of the Egernia whitii (Lacertilia: Scincidae) species group. AB - We assembled a molecular phylogeny for the arid-zone members of the Egernia whitii species group to test Pianka's [Zoogeography and speciation of Australian desert lizards: an ecological perspective, Copeia (1972) 127-145] hypothesis that habitat specificity to the three major arid-zone vegetation communities is the primary cause of lizard speciation within the arid interior of Australia. This hypothesis predicts that species should exhibit phylogeographic structuring concordant with the major arid-zone vegetation types. Sequence data were obtained from four of the five arid-zone members of the E. whitii species group, and from across the ranges of the ecologically generalized E. inornata and E. multiscutata and the more specialized E. striata. We targeted a fragment (696 base pair (bp)) of the mitochondrial genome comprising the 3' half of the ND4 gene. We analysed the data using parsimony, maximum likelihood and Bayesian methods. Our phylogeny confirms the monophyly of the arid-zone members of the species group, although the phylogenetic relationships among species were not fully resolved. Although our topology does not support the recognition of the existing subspecies within E. multiscutata, there is a substantial phylogeographic break between South Australian/Victorian (Clade 1) and Western Australian (Clade 2) populations. We found considerable phylogeographic structure within E. inornata, with six major clades identified. However, these clades were not concordant with the distribution of habitat types in the arid-zone. Phylogeographic structure was also observed in the more specialized E. striata, although our analysis revealed close phylogenetic affinities between the sympatric species E. striata and E. kintorei. Shimodaira-Hasegawa topology tests were equivocal in regard to whether the phylogeographic structure within E. striata was in accordance with Pianka's predictions. Although our data failed to provide strong support for the suggestion that ecological and habitat factors are responsible for the diversification of arid-zone lizards, most E. inornata and E. striata populations had similar habitats, indicating that adaptation to particular habitats may have some role in the speciation of lizards in the Australian arid-zone. PMID- 15522788 TI - Molecular phylogenetics and diversification of the genus Sporophila (Aves: Passeriformes). AB - The evolutionary affinities within and among many groups of nine-primaried oscines remain unresolved. One such group is Sporophila, a large genus of New World tanager-finches. Our study focused particularly on clarifying the relationship between this genus and a closely related one, Oryzoborus, and on examining the phylogenetic affinities of the "capuchinos," a group of 11 Sporophila species that share a similar male plumage coloration pattern. Our phylogenetic analyses, based on 498 bp of mitochondrial DNA sequence, indicated that: (1) Oryzoborus is embedded within a well-supported clade containing all Sporophila species, which strongly suggests that both genera should be merged, (2) the species of capuchinos comprise a monophyletic group, implying that the plumage patterns common to all probably arose only once, and (3) the capuchinos clade is comprised of two sub-clades, one including two species that are distributed in northern South America and the other one containing eight species that are present south of the Amazon River. Mean sequence divergence among the southern capuchinos species was extremely low, suggesting a rapid radiation within the last half-million years that may be related to the high level of sexual selection present in the genus and might have been promoted by marine ingressions and egressions that occurred in some southern coastal regions of South America in the Late Pleistocene. PMID- 15522789 TI - Heterogeneous evolution of the Myc-like Anthocyanin regulatory gene and its phylogenetic utility in Cornus L. (Cornaceae). AB - Anthocyanin is a major pigment in vegetative and floral organs of most plants and plays an important role in plant evolution. The anthocyanin regulatory genes are responsible for regulating transcription of genes in the anthocyanin synthetic pathway. To assess evolutionary significance of sequence variation and evaluate the phylogenetic utility of an anthocyanin regulatory gene, we compared nucleotide sequences of the myc-like anthocyanin regulatory gene in the genus of dogwoods (Cornus: Cornaceae). Phylogenetic analyses demonstrate that the myc-like anthocyanin regulatory gene has potential as an informative phylogenetic marker at different taxonomic levels, depending on the data set considered (DNA or protein sequences) and regions applied (exons or introns). Pairwise nonsynonymous and synonymous substitution rate tests and codon-based substitution models were applied to characterize variation and to identify sites under diversifying selection. Mosaic evolution and heterogeneous rates among different domains and sites were detected. PMID- 15522790 TI - Phylogeny of Lasius ants based on mitochondrial DNA and morphology, and the evolution of social parasitism in the Lasiini (Hymenoptera: Formicidae). AB - Phylogeny of ants of the tribe Lasiini (Lasius, Acanthomyops, Prenolepis, Euprenolepis, Paratrechina, Pseudolasius, and Myrmecocystus) was analysed using 81 morphological, ecological, and behavioural characters (for 41 species) and mitochondrial DNA sequences (COI, COII, tRNA-Leu; for 19 species). The free living subgenus Lasius s. str. is paraphyletic with respect to the rest of genus; the traditional "genus" Acanthomyops should be considered a part of Lasius s. lat.; free-living subgenus Cautolasius is a member of the clade of socially parasitic Lasius ants (=Chtonolasius+Acanthomyops+Austrolasius+Dendrolasius). The tree topology is congruent with two alternative scenarios of origin of the temporary social parasitism: (i) a single origin of the parasitic strategy in a derived subclade of Lasius and a secondary loss of this trait in Cautolasius, (ii) a parallel origin of the social parasitism within the clade of hypogeic Lasius ants (in Chtonolasius, and in Acanthomyops+Dendrolasius+Austrolasius). Emery's rule in the strict sense does not apply to this group because most parasites exploit any ecologically available, even phylogenetically distant host species. The parasitic strategy in Lasius could have originated from the aggressive interactions between cofounding queens during pleometric colony founding and/or from the secondary queen adoption. PMID- 15522791 TI - Ribosomal protein-sequence block structure suggests complex prokaryotic evolution with implications for the origin of eukaryotes. AB - Amino acid sequence alignments of orthologous ribosomal proteins found in Bacteria, Archaea, and Eukaryota display, relative to one another, an unusual segment or block structure, with major evolutionary implications. Within each of the prokaryotic phylodomains the sequences exhibit substantial similarity, but cross-domain alignments break up into (a) universal blocks (conserved in both phylodomains), (b) bacterial blocks (unalignable with any archaeal counterparts), and (c) archaeal blocks (unalignable with any bacterial counterparts). Sequences of those eukaryotic cytoplasmic riboproteins that have orthologs in both Bacteria and Archaea, exclusively match the archaeal block structure. The distinct blocks do not correlate consistently with any identifiable functional or structural feature including RNA and protein contacts. This phylodomain-specific block pattern also exists in a number of other proteins associated with protein synthesis, but not among enzymes of intermediary metabolism. While the universal blocks imply that modern Bacteria and Archaea (as defined by their translational machinery) clearly have had a common ancestor, the phylodomain-specific blocks imply that these two groups derive from single, phylodomain-specific types that came into existence at some point long after that common ancestor. The simplest explanation for this pattern would be a major evolutionary bottleneck, or other scenario that drastically limited the progenitors of modern prokaryotic diversity at a time considerably after the evolution of a fully functional translation apparatus. The vast range of habitats and metabolisms that prokaryotes occupy today would thus reflect divergent evolution after such a restricting event. Interestingly, phylogenetic analysis places the origin of eukaryotes at about the same time and shows a closer relationship of the eukaryotic ribosome-associated proteins to crenarchaeal rather than euryarchaeal counterparts. PMID- 15522792 TI - Temporal, spatial, and ecological modes of evolution of Eurasian Mus based on mitochondrial and nuclear gene sequences. AB - We sequenced mitochondrial (cytochrome b, 12S rRNA) and nuclear (IRBP, RAG1) genes for 17 species of the Old World murine genus Mus, drawn primarily from the Eurasian subgenus Mus. Phylogenetic analysis of the newly and previously available sequences support recognition of four subgenera within Mus (Mus, Coelomys, Nannomys, and Pyromys), with an unresolved basal polytomy. Our data further indicate that the subgenus Mus contains three distinct 'species groups': (1) a Mus booduga Species Group, also including Mus terricolor and Mus fragilicauda (probably also Mus famulus); (2) a Mus cervicolor Species Group, also including Mus caroli and Mus cookii; and (3) a Mus musculus Species Group, also including Mus macedonicus, Mus spicilegus, and Mus spretus. Species diversity in Eurasian Mus is probably explicable in terms of several phases of range expansion and vicariance, and by a propensity within the group to undergo biotope transitions. IRBP and RAG1 molecular clocks for Mus date the origin of subgenera to around 5-6 mya and the origin of Species Groups within subgenus Mus to around 2-3 mya. The temporal pattern of evolution among Eurasian Mus is more complex than that within the Eurasian temperate genus Apodemus. PMID- 15522793 TI - Molecular phylogeny of the speciose vole genus Microtus (Arvicolinae, Rodentia) inferred from mitochondrial DNA sequences. AB - Voles of the genus Microtus represent one of the most speciose mammalian genera in the Holarctic. We established a molecular phylogeny for Microtus to resolve contentious issues of systematic relationships and evolutionary history in this genus. A total of 81 specimens representing ten Microtus species endemic to Europe as well as eight Eurasian, six Asian and one Holarctic species were sequenced for the entire cytochrome b gene (1140 bp). A further 25 sequences were retrieved from GenBank, providing data on an additional 23, mainly Nearctic, Microtus species. Phylogenetic analysis of these 48 species generated four well supported monophyletic lineages. The genus Chionomys, snow voles, formed a distinct and well-supported lineage separate from the genus Microtus. The subgenus Microtus formed the strongest supported lineage with two sublineages displaying a close relationship between the arvalis species group (common voles) and the socialis species group (social voles). Monophyly of the Palearctic pitymyid voles, subgenus Terricola, was supported, and this subgenus was also subdivided into two monophyletic species groups. Together, these groupings clarify long-standing taxonomic uncertainties in Microtus. In addition, the "Asian" and the Nearctic lineages reported previously were identified although the latter group was not supported. However, relationships among the main Microtus branches were not resolved, suggesting a rapid and potentially simultaneous radiation of a widespread ancestor early in the history of the genus. This and subsequent radiations discernible in the cytochrome b phylogeny, show the considerable potential of Microtus for analysis of historical and ecological determinants of speciation in small mammals. It is evident that speciation is an ongoing process in the genus and that the molecular data provides a vital insight into current species limits as well as cladogenic events of the past. PMID- 15522794 TI - A genealogical view of chromosomal evolution and species delimitation in the Drosophila virilis species subgroup. AB - Chromosomal arrangement was a historically important character used for defining taxonomic boundaries. The Drosophila virilis species group exhibits a series of chromosomal rearrangements, and the resulting differences among karyotypes were primary characters originally used to define taxa within the group. However, some chromosomally divergent forms have not been sufficiently resolved in phylogenetic reconstructions of DNA sequences from several nuclear genes. Sequences of mitochondrial regions have the potential for finer-scale resolution of closely related taxa; therefore, sequences of two mitochondrial genes were used to examine phylogenetic relationships within the chromosomally variable virilis subgroup. Sequences were obtained from multiple strains of the Palearctic species, D. virilis and D. lummei, and the Nearctic species, D. novamexicana and two chromosomal forms of D. americana. Analyses support the recent emergence of the different chromosomal forms in North America. However, none of these chromosomally divergent forms exhibit reciprocal monophyly of their mtDNA sequences, which is the requirement for attaining genealogical species status. PMID- 15522795 TI - Phylogenetic analysis of the nuclear alcohol dehydrogenase (Adh) gene family in Carex section Acrocystis (Cyperaceae) and combined analyses of Adh and nuclear ribosomal ITS and ETS sequences for inferring species relationships. AB - We analyzed sequence variation for the alcohol dehydrogenase (Adh) gene family in Carex section Acrocystis (Cyperaceae) to reconstruct Adh gene trees for Acrocystis species and to characterize the structure of the Adh gene family in Carex. Two Adh loci were included with ITS and ETS sequences in a combined Bayesian inference analysis of Carex section Acrocystis to gain a better understanding of species relationships in the section. In addition, we comment on how the results presented here contribute to our knowledge of the birth-death process of the Adh gene family in angiosperms. It appears that the structure of the Adh gene family in Carex is complex with possibly six loci present in the gene family. Additionally, variation among Acrocystis species within loci is quite low, and there is little phylogenetic resolution in the individual datasets. Bayesian inference analysis of the combined ITS, ETS, Adh1, and Adh2 datasets resulted in a moderately well-supported phylogenetic hypothesis of relationships in the section which is discussed in relation to previous hypotheses of relationships. PMID- 15522796 TI - Detecting phylogenetic incongruence using BIONJ: an improvement of the ILD test. AB - The problem of testing for congruence between phylogenetic data has long been debated among phylogeneticists, but reaches a critical point with the availability of large amount of biological sequences. Notably in prokaryotes, where the amount of lateral transfers is believed to be important, the inference of phylogenies using multiple genes requires testing for incongruence before concatenating the genes. On another scale, incongruence tests can be used to detect recombination points within single gene alignments. The incongruence length difference test (ILD), based on parsimony, has been proved to be useful for finding incongruent data sets, but its application remains limited to small data sets for computational time reasons. Here, we have adapted the principle of ILD to the BIONJ algorithm. This algorithm is based on a tree length minimisation criterion and is suitable to replace parsimony in this test when used with uncorrected distance (model-free approach). We show that this new test, ILD BIONJ, while being much faster, is often more accurate than the ILD test, especially when the alignments compared are simulated under different evolutionary models. PMID- 15522797 TI - Phylogenetic relationships within mammalian order Carnivora indicated by sequences of two nuclear DNA genes. AB - Phylogenetic relationships among 37 living species of order Carnivora spanning a relatively broad range of divergence times and taxonomic levels were examined using nuclear sequence data from exon 1 of the IRBP gene (approximately 1.3 kb) and first intron of the TTR gene (approximately 1 kb). These data were used to analyze carnivoran phylogeny at the family and generic level as well as the interspecific relationships within recently derived Felidae. Phylogenetic results using a combined IRBP+TTR dataset strongly supported within the superfamily Califormia, the red panda as the closest lineage to procyonid-mustelid (i.e., Musteloidea) clade followed by pinnipeds (Otariidae and Phocidae), Ursidae (including the giant panda), and Canidae. Four feliform families, namely the monophyletic Herpestidae, Hyaenidae, and Felidae, as well as the paraphyletic Viverridae were consistently recovered convincingly. The utilities of these two gene segments for the phylogenetic analyses were extensively explored and both were found to be fairly informative for higher-group associations within the order Carnivora, but not for those of low level divergence at the species level. Therefore, there is a need to find additional genetic markers with more rapid mutation rates that would be diagnostic at deciphering relatively recent relationships within the Carnivora. PMID- 15522798 TI - Labile male morphology and intraspecific male polymorphism in the Philotrypesis fig wasps. AB - We investigate the evolution of male morphology in the fig wasps belonging to the genus Philotrypesis (Chalcidoidea, Sycorectinae). We first reconstruct the phylogenetic relationships of Philotrypesis associated with African figs using nuclear and mitochondrial DNA. We then determine male morphotypes in the species included in our phylogeny and show that intraspecific polymorphism is common. Most species present two types of males and some species have up to three types. These morphotypes are believed to represent alternative mating tactics: some males show morphological adaptations to fighting, others are winged dispersers and others are small sneakers. Mapping out these variations onto our phylogeny reveals that the combination of morphs changes randomly along the branches of the tree. Both parsimony and likelihood approaches indicate that there has been at least one transition from dimorphism to trimorphism, several gains and losses of the small morph and two independent acquisitions of the winged morph. Using maximum likelihood analyses of character evolution, we estimate transition rates for each morph and show that the evolution of each type of morph are not correlated and that forward and backward transition rates are not significantly different. Our results altogether suggest that male morphology is evolutionary labile, it responds quickly to selection imposed by the mating environment. This study, also suggests that seemingly complex phenotypes, such as winged males, can evolve several times and can even be recreated after having been lost. PMID- 15522799 TI - Phylogeny and biogeography of the Malagasy and Australasian rainbowfishes (Teleostei: Melanotaenioidei): Gondwanan vicariance and evolution in freshwater. AB - Phylogenetic relationships of the Malagasy and Australasian rainbowfishes are investigated using 4394 characters derived from five mitochondrial genes (12S, 16S, tRNA-Valine, ND5, and COI), three nuclear genes (28S, histone H3, and TMO 4c4), and 102 morphological transformations. This study represents the first phylogenetic analysis of the endemic Malagasy family Bedotiidae and includes a nearly complete taxonomic review of all nominal species, as well as numerous undescribed species. Simultaneous analysis of the molecular and morphological datasets results in two equally most parsimonious trees. Results indicate that Bedotiidae (Bedotia+Rheocles) and Bedotia are monophyletic, whereas Rheocles is paraphyletic with the inclusion of two recently described species from northeastern Madagascar, R. vatosoa, and R. derhami. Rheocles vatosoa and R. derhami are sister taxa, and this clade is recovered as the sister group to Bedotia. The remaining species of Rheocles are not sexually dimorphic and comprise a clade that is recovered as the sister group to Bedotia+(R. derhami+R. vatosoa), all of which are sexually dichromatic, and sexually dimorphic for pigmentation and fin development. Three geographically distinct clades are recovered within Bedotia, one comprising species with distributions ranging from mid- to southeastern Madagascar, another including species restricted to eastern drainages north of the Masoala Peninsula, and a third comprising taxa with distributions extending from the Masoala Peninsula south to the Ivoloina River. The Australian/New Guinean melanotaeniids are monophyletic and are recovered as the sister group to Bedotiidae. The Australasian Telmatherinidae and Pseudomugilidae comprise a clade that is recovered as the sister group to the Melanotaeniidae-Bedotiidae clade. This sister-group relationship between Malagasy bedotiids and a clade restricted to Australia-New Guinea, and the absence of a close relationship between bedotiids and African or Mascarene atheriniforms, is congruent with the break-up of Gondwana, not a scenario reliant on Cenozoic trans oceanic dispersal. Finally, results of the phylogenetic analysis indicate that Atheriniformes is polyphyletic and further corroborate recent morphological hypotheses, which have recovered Bedotiidae in a derived position within Atherinoidei. PMID- 15522800 TI - Molecular phylogeny and biogeography of woolly flying squirrel (Rodentia: Sciuridae), inferred from mitochondrial cytochrome b gene sequences. AB - To investigate the genetic diversity between the populations of woolly flying squirrels (Eupetaurus) from the eastern and western extremes of the Himalayas, partial mitochondrial cytochrome b gene sequences (390-810 bp) that were determined from the museum specimens were analyzed using maximum parsimony (MP) and maximum likelihood (ML) methods. The molecular data reveal that the two specimens that were collected in northwestern Yunnan (China) are members of the genus Eupetaurus. Reconstructed phylogenetic relationships show that the populations of Eupetaurus in the eastern and western extremes of the Himalayas are two distinct species with significant genetic differences (12%) and diverged about 10.8 million years ago. Eupetaurus is significantly different from Petaurista and Pteromys. The level of estimated pairwise-sequence divergence observed between Eupetaurus and Petaurista or Pteromys is greater than that observed between Eupetaurus and Trogopterus, Belomys, Glaucomys, or Hylopetes. Considering the divergence time of the two Eupetaurus groups, the glaciations and the uplift of the Himalayas and Qinghai-Tibet plateau during the Pliocene Pleistocene period might be the major factors affecting the present distribution of Eupetaurus along the Himalayas. PMID- 15522801 TI - Molecular phylogeny of the fern genus Elaphoglossum (Elaphoglossaceae) based on chloroplast non-coding DNA sequences: contributions of species from the Indian Ocean area. AB - We performed a phylogenetic analysis of the fern genus Elaphoglossum using two non-coding chloroplast spacers: trnL-trnF and rps4-trnS. The sampling includes 123 species, of which 80 have not been previously sequenced, and for the first time includes species from Africa and the Indian Ocean area. The results of this expanded study largely agree with an earlier molecular study based on a smaller group of neotropical species and with the morphology-based classification of Mickel and Atehortua. We found, however, that some infrageneric groups such as section Elaphoglossum are not monophyletic. Besides section Elaphoglossum pro parte, we recognize six sections: two new monospecific, unnamed sections, and the previously established sections Lepidoglossa, Squamipedia, Amygdalifolia, and "Subulate-scaled clade." We divide the subulate-scaled clade into subsection Setosa (hydathodes present) and Polytrichia (hydathodes absent), and section Elaphoglossum is divided into subsections Platyglossa and Pachyglossa, two groups that do not appear to be supported by any single morphological character. In general, however, the main clades are supported by morphology. Finally, we discuss the species of the Indian Ocean region and their affinities with the neotropical ones. Out of the 11 species pairs postulated by Moran and Smith on the basis of morphology, two are well supported (E. eximium-E. aubertii; E. piloselloides-E. spatulatum) and three are not supported (E. ciliatum-E. humbertii; E. muscosum-E. poolii; E. paleaceum-E. deckenii), and two remain unresolved (E. erinaceum-E. hybridum; E. glabellum-E. acrostichoides) because our molecular markers were not variable enough. Four species pairs could not be tested because specimens were lacking. Unsupported species pairs are best interpreted as morphological convergences. Two additional species pairs are proposed: E. cuspidatum-E. succisaefolium; E. doanense-E. hornei. Placement of the species from the Indian Ocean suggests that at least 13 long-distance dispersal events occurred between the Neotropics and the Indian Ocean-Africa. PMID- 15522802 TI - Phylogeny and phylogeography of Old World fruit bats in the Cynopterus brachyotis complex. AB - Taxonomic relationships within the Old World fruit bat genus, Cynopterus, have been equivocal for the better part of a century. While nomenclature has been revised multiple times on the basis of phenotypic characters, evolutionary relationships among taxa representing the entire geographic range of the genus have not been determined. We used mitochondrial DNA sequence data to infer phylogenetic relationships among the three most broadly distributed members of the genus: C. brachyotis, C. horsfieldi, and C. sphinx, and to assess whether C. brachyotis represents a single widespread species, or a complex of distinct lineages. Results clearly indicate that C. brachyotis is a complex of lineages. C. sphinx and C. horsfieldi haplotypes formed monophyletic groups nested within the C. brachyotis species complex. We identified six divergent mitochondrial lineages that are currently referred to C. brachyotis. Lineages from India, Myanmar, Sulawesi, and the Philippines are geographically well-defined, while in Malaysia two lineages, designated Sunda and Forest, are broadly sympatric and may be ecologically distinct. Demographic analyses of the Sunda and Forest lineages suggest strikingly different population histories, including a recent and rapid range expansion in the Sunda lineage, possibly associated with changes in sea levels during the Pleistocene. The resolution of the taxonomic issues raised in this study awaits combined analysis of morphometric characters and molecular data. However, since both the Indian and Malaysian Forest C. brachyotis lineages are apparently ecologically restricted to increasingly fragmented forest habitat, we suggest that reevaluation of the conservation status of populations in these regions should be an immediate goal. PMID- 15522803 TI - Myostatin rapid sequence evolution in ruminants predates domestication. AB - Myostatin (GDF-8) is a negative regulator of skeletal muscle development. This gene has previously been implicated in the double muscling phenotype in mice and cattle. A systematic analysis of myostatin sequence evolution in ruminants was performed in a phylogenetic context. The myostatin coding sequence was determined from duiker (Sylvicapra grimmia caffra), eland (Taurotragus derbianus), gaur (Bos gaurus), ibex (Capra ibex), impala (Aepyceros melampus rednilis), pronghorn (Antilocapra americana), and tahr (Hemitragus jemlahicus). Analysis of nonsynonymous to synonymous nucleotide substitution rate ratios (Ka/Ks) indicates that positive selection may have been operating on this gene during the time of divergence of Bovinae and Antilopinae, starting from approximately 23 million years ago, a period that appears to account for most of the sequence difference between myostatin in these groups. These periods of positive selective pressure on myostatin may correlate with changes in skeletal muscle mass during the same period. PMID- 15522804 TI - Genetic diversity and biogeography of Cunninghamia konishii (Cupressaceae), an island species in Taiwan: a comparison with Cunninghamia lanceolata, a mainland species in China. AB - Luanta-fir (Cunninghamia konishii), an endemic to Taiwan, is an outcrossing, long lived conifer. Populations of C. konishii are generally fragmented due to a once high intensity of timber exploitation. C. konishii and Cunninghamia lanceolata are two sibling taxa constituting derivative-progenitor species relationship. The amount of genetic variations within and between 11 and 10 populations of C. konishii and C. lanceolata, respectively, were assessed using amplified fragment length polymorphism (AFLP) markers in this report. Three AFLP primer pairs generated a total of 357 and 226 markers for C. konishii and C. lanceolata samples, of which 56.1 and 65.3% are polymorphic, respectively. Analysis of molecular variance indicates a 4.78% variation between C. konishii and C. lanceolata. A relatively high value of genetic variation (24.60%) was apportioned between the populations of C. konishii. In contrast, a lower divergence value (12.21%) between populations was found for C. lanceolata. The population with the highest genetic diversity was found in Nantou County, which concurred with the results of many other tree species investigated in Taiwan. The estimates of the number of migrants between populations (Nm), obtained from population pair-wise PhiST, suggest that gene flow in C. konishii is efficient in some adjacent populations but is restricted in the rest. Individual UPGMA tree, generated based on AFLP markers, suggests six evolutionary lineages for C. konishii. All evolutionary lineages of C. konishii were derived from C. lanceolata. In conclusion, the migration patterns of Cunninghamia from mainland China may have been established following multiple sources, migrant-pools, long-distance dispersal events, and via different directions. PMID- 15522805 TI - Phylogenetic relationships and biogeography of the genus Chondrostoma inferred from mitochondrial DNA sequences. AB - A phylogeny of the species of the nase genus Chondrostoma was constructed from a complete mitochondrial cytochrome b gene (1140 bp). Molecular phylogeny was used to revise the current systematics of this group, and to infer a biogeographical model of the Mediterranean area during the Cenozoic period. We confirmed the monophyly of the genus Chondrostoma, and defined seven different lineages within it: Polylepis, Arcasii, Lemmingii, Toxostoma, Nasus, C. genei, and C. soetta. The separation of main lineages within Chondrostoma occurred in the Middle-Upper Miocene, approximately 11 million years ago, while the greatest species radiation took place in the Pliocene close to the time the current drainages system were created. It is unlikely that this genus experienced an extensive dispersal during the Messinian, in the Lago-Mare Phase. Given the level of current knowledge, a biogeographical model constructed on the basis of vicariant events seems more realistic than does a dispersalist model. PMID- 15522806 TI - Intragenomic variation of the rDNA internal transcribed spacers in sponges (Phylum Porifera): implications for phylogenetic studies. AB - The internal transcribed spacer regions (ITS1 and ITS2) of the tandemly repeated nuclear ribosomal DNA clusters are frequently used as markers for fine scale analyses in diverse animals. In certain taxa, ITS is nearly exclusively used for population level or inter-specific studies, despite the frequent presence of divergent paralogs within individual genomes that can be phylogenetically misleading. For the first time we survey diverse marine sponges to determine the extent and phylogenetic implications of intragenomic polymorphisms (IGPs) exhibited at their ITS loci. We discover that the extent of IGP varies greatly between taxa (with most taxa exhibiting very few) and cannot be predicted by taxonomy. Furthermore, we demonstrate that ITS can be phylogenetically informative between species when moderate levels of IGPs are detected, but that ITS paralogy can interfere with population level studies. We caution against the routine use of ITS in phylogenetic studies of sponges without (1) screening for IGPs in specimens from every population sampled; (2) including all divergent paralogs in phylogenetic analyses; (3) testing ITS data using other single-copy, unlinked loci (such as nuclear introns). PMID- 15522807 TI - Historical biogeography of Mediterranean trout. AB - Complete sequencing of the mitochondrial control region was used to describe phylogenetic relationships of brown trout populations (Salmo trutta) in the Mediterranean river basins of Iberia and to review the historical biogeography of trout from the Mediterranean regions. Phylogenetic relationships among trout lineages suggested that the Danubian one is the most ancestral, in accordance with the eastern origin of most of the European freshwater fish species. Nested clade and mismatch analyses suggested that the present distribution of haplotypes of the Adriatic and Mediterranean lineages resulted from population expansions originated, respectively, from central and western Europe, which favoured extensive secondary contacts between lineages. Reduced diversity detected within 50% of the analysed populations and large intrabasin differentiation indicated restricted gene flow in post-glacial periods. PMID- 15522808 TI - Phylogeny, biogeography, and the evolution of life-history traits in Leucadendron (Proteaceae). AB - Leucadendron is a moderately large genus of Proteaceae almost entirely restricted to the Cape Floristic Region of southern Africa. The genus is unusual in being dioecious and sexually dimorphic. ITS sequence data were obtained from 62 of the 96 currently recognized taxa (85 species and 11 subspecies). Phylogenetic analyses were conducted under Maximum Likelihood and parsimony and resolved nine groups of species with varying degrees of bootstrap support, but relationships between these groups are largely unsupported. The phylogeny conflicts with the current taxonomic arrangement, which is based mainly on fruit morphology. The two sections of the genus, Alatosperma and Leucadendron, and several subsections within these sections, are resolved as non-monophyletic. This means that taxonomically important characters (such as fruit shape) have evolved multiple times, as the species with nut-like fruit (resolved into two of the nine groups) appear to have evolved independently from ancestors with winged fruit. Based on the topology obtained, the life history traits of anemophily, myrmechochory, and re-sprouting have also originated multiple times. Dispersal-Vicariance (DIVA) analysis suggests that the genus had an ancestral area in the Karoo Mountain and Southeastern phytogeographic centres of endemism in the southwestern Cape. PMID- 15522809 TI - Phylogenetic and biogeographic diversification of Rhus (Anacardiaceae) in the Northern Hemisphere. AB - Sequences of internal transcribed spacers (ITS) of nuclear ribosomal DNA, the chloroplast ndhF gene, and chloroplast trnL-F regions (trnL intron, and trnL [UAA] 3' exon-trnF [GAA] intergenic spacer) were used for phylogenetic analyses of Rhus, a genus disjunctly distributed in Asia, Europe, Hawaii, North America, and Northern Central America. Both ITS and cpDNA data sets support the monophyly of Rhus. The monophyly of subgenus Rhus was suggested by the combined cpDNA and ITS data, and largely supported in the cpDNA data except that Rhus microphylla of subgenus Lobadium was nested within it. The monophyly of subgenus Lobadium was strongly supported in the ITS data, whereas the cpDNA data revealed two main clades within the subgenus, which formed a trichotomy with the clade of subgenus Rhus plus R. microphylla. The ITS and cpDNA trees differ in the positions of Rhus michauxii, R. microphylla, and Rhus rubifolia, and hybridization may have caused this discordance. Fossil evidence indicates that Rhus dates back to the early Eocene. The penalized likelihood method was used to estimate divergence times, with fossils of Rhus subgenus Lobadium, Pistacia and Toxicodendron used for age constraints. Rhus diverged from its closest relative at 49.1+/-2.1 million years ago (Ma), the split of subgenus Lobadium and subgenus Rhus was at 38.1+/-3.0 Ma. Rhus most likely migrated from North America into Asia via the Bering Land Bridge during the Late Eocene (33.8+/-3.1 Ma). Rhus coriaria from southern Europe and western Asia diverged from its relatives in eastern Asia at 24.4+/-3.2 Ma. The Hawaiian Rhus sandwicensis diverged from the Asian Rhus chinensis at 13.5+/-3.0 Ma. Subgenus Lobadium was inferred to be of North American origin. Taxa of subgenus Lobadium then migrated southward to Central America. Furthermore, we herein make the following three nomenclatural combinations: (1) Searsia leptodictya (Diels) T. S. Yi, A. J. Miller and J. Wen, comb. nov., (2) Searsia pyroides (A. Rich.) T. S. Yi, A. J. Miller and J. Wen, comb. nov., and (3) Searsia undulata (Jacq.) T. S. Yi, A. J. Miller and J. Wen, because our analyses support the segregation of Searsia from Rhus. PMID- 15522810 TI - Evolution and phylogeny of old world deer. AB - The phylogenetic pattern and timing of the radiation of Old World deer was determined based on the complete mitochondrial cytochrome b gene from 33 Cervinae taxa. Using rooted and unrooted phylogenies derived from distinct theoretical approaches, strong support was achieved for monophyly of the Old World deer with muntjacs as sister group as well as for the divergence of at least three distinct genera: Rucervus, Dama, and Cervus. The latter clade comprises what have previously been regarded as the genera or subgenera Panolia, Rusa, Cervus, Sika, and probably Przewalskium. Our data also consistently confirmed paraphyly of nominate C. elaphus and did not support the monophyly of Axis. We used these molecular phylogenies to assess the homoplastic evolution of morphological, geographical, ecological, and selected behavioural character state differences within the Cervinae. Reliable fossil calibrations, large molecular data sets, and improved dating methods are shaping a molecular time scale for the evolutionary radiation of Old World deer that occurred at the Miocene/Pliocene transition and is largely compatible with existing palaeontological evidence. Using node ages estimated from sequence data, we estimated an average per-lineage diversification rate of 0.51+/-0.1 species per million years (my) over roughly the last 6 mya. PMID- 15522811 TI - Molecular phylogeny of the tribe Bovini (Bovidae, Bovinae) and the taxonomic status of the Kouprey, Bos sauveli Urbain 1937. AB - The kouprey is a very rare bovid species of the Indochinese peninsula, and no living specimen has been described for a long time, suggesting that it is possibly extinct. Its systematic position within the tribe Bovini remains confused since the analyses of morphological characters have led to several conflicting hypotheses. Some authors have also suggested that it could be a hybrid species produced by the crossing of the banteng with gaur, zebu, or water buffalo. Here we performed a molecular phylogeny of the tribe Bovini to determine the taxonomic status of the kouprey. DNA was extracted from the holotype specimen preserved in the MNHN collections. Phylogenetic analyses were carried out on a matrix including all the taxonomic diversity described in the tribe Bovini, and 2065 nucleotide characters, representing three different markers, i.e., the promotor of the lactoferrin and two mitochondrial genes (cytochrome b and subunit II of the cytochrome c oxidase). The results show that the kouprey belongs to the subtribe Bovina, and that three different clades can be evidenced into this group: the first includes the domestic ox, zebu, and European bison; the second incorporates the yak and American bison; and the third contains the kouprey, banteng and gaur. All hypotheses involving hybridization for the origin of the kouprey can be rejected, confirming that it is a real wild species. Molecular datings and biogeographic inferences suggest that the kouprey diverged from banteng and gaur during the Plio-Pleistocene of Asia. In addition, several molecular signatures were detected in the cytochrome b gene, permitting a molecular identification of the kouprey. We propose a conservation project based on a molecular taxonomy approach for tracking the kouprey in Indochina in order to determine whether some populations still survive in the wild. PMID- 15522812 TI - Convergent evolution of strigiform and caprimulgiform dark-activity is supported by phylogenetic analysis using the arylalkylamine N-acetyltransferase (Aanat) gene. AB - Alternative hypotheses propose the sister order of owls (Strigiformes) to be either day-active raptors (Falconiformes) or dark-active nightjars and allies (Caprimulgiformes). In an effort to identify molecular characters distinguishing between these hypotheses we examined a gene, arylalkylamine N-acetyltransferase (Aanat), potentially associated with the evolution of avian dark-activity. Partial Aanat coding sequences, and two introns, were obtained from the genomic DNA of 16 species: Strigiformes (four species), Falconiformes (four species), Caprimulgiformes (five species), with outgroups: Ciconiiformes (one species), Passeriformes (one species), and Apterygiformes (one species). Phylogenetic trees derived from aligned, evolutionarily conserved Aanat regions did not consistently recover clades corresponding to orders Strigiformes and Falconiformes but did place a caprimulgiform clade more distant from the strigiform and falconiform species than the latter two groups are to each other. This finding was supported by spectral analysis. The taxonomic distribution of seven intronic indels is consistent with the Aanat derived phylogenetic trees and supports conventional family-level groupings within both Strigiformes and Caprimulgiformes. The phylogenetic analyses also indicate that Caprimulgiformes is a polyphyletic grouping. In conclusion the data support, but do not conclusively prove, the proposal that Falconiformes is the sister order to Strigiformes and therefore, that the dark-activity characteristic of Strigiformes and Caprimulgiformes arose by convergent evolution. PMID- 15522813 TI - Rabbits, if anything, are likely Glires. AB - Rodentia (e.g., mice, rats, dormice, squirrels, and guinea pigs) and Lagomorpha (e.g., rabbits, hares, and pikas) are usually grouped into the Glires. Status of this controversial superorder has been evaluated using morphology, paleontology, and mitochondrial plus nuclear DNA sequences. This growing corpus of data has been favoring the monophyly of Glires. Recently, Misawa and Janke [Mol. Phylogenet. Evol. 28 (2003) 320] analyzed the 6441 amino acids of 20 nuclear proteins for six placental mammals (rat, mouse, rabbit, human, cattle, and dog) and two outgroups (chicken and xenopus), and observed a basal position of the two murine rodents among the former. They concluded that "the Glires hypothesis was rejected." We here reanalyzed [loc. cit.] data set under maximum likelihood and Bayesian tree-building approaches, using phylogenetic models that take into account among-site variation in evolutionary rates and branch-length variation among proteins. Our observations support both the association of rodents and lagomorphs and the monophyly of Euarchontoglires (=Supraprimates) as the most likely explanation of the protein alignments. We conducted simulation studies to evaluate the appropriateness of lissamphibian and avian outgroups to root the placental tree. When the outgroup-to-ingroup evolutionary distance increases, maximum parsimony roots the topology along the long Mus-Rattus branch. Maximum likelihood, in contrast, roots the topology along different branches as a function of their length. Maximum likelihood appears less sensitive to the "long branch attraction artifact" than is parsimony. Our phylogenetic conclusions were confirmed by the analysis of a different protein data set using a similar sample of species but different outgroups. We also tested the effect of the addition of afrotherian and xenarthran taxa. Using the linearized tree method, [loc. cit.] estimated that mice and rats diverged about 35 million years ago. Molecular dating based on the Bayesian relaxed molecular clock method suggests that the 95% credibility interval for the split between mice and rats is 7-17 Mya. We here emphasize the need for appropriate models of sequence evolution (matrices of amino acid replacement, taking into account among-site rate variation, and independent parameters across independent protein partitions) and for a taxonomically broad sample, and conclude on the likelihood that rodents and lagomorphs together constitute a monophyletic group (Glires). PMID- 15522814 TI - Characterizing regions of ambiguous alignment caused by the expansion and contraction of hairpin-stem loops in ribosomal RNA molecules. PMID- 15522815 TI - Coadaptive evolution in cytochrome c oxidase: 9 of 13 subunits show accelerated rates of nonsynonymous substitution in anthropoid primates. PMID- 15522816 TI - Role of FATP in parenchymal cell fatty acid uptake. AB - Long-chain fatty acids (LCFAs) represent key metabolites for energy generation and storage. Transport and metabolism of LCFA are believed to be regulated by membrane-associated proteins that bind and transport LCFA. Identifying the postulated fatty acid transporters is of considerable interest since altered fatty acid uptake has been implicated in disease such as insulin resistance and obesity. Recently, a family of membrane associated proteins, termed fatty acid transport proteins (FATPs), have been described that enhance uptake of LCFAs. Until today, six members of this family, designated FATP1-6, have been characterized. This review will focus on FATP structure, expression patterns, regulation, mechanism of transport and clinical implications. PMID- 15522817 TI - The novel pathway for ketodiene oxylipin biosynthesis in Jerusalem artichoke (Helianthus tuberosus) tubers. AB - The new route of the plant lipoxygenase pathway, directed specifically towards the ketodiene formation, was detected during in vitro experiments with Jerusalem artichoke (Helianthus tuberosus) tubers. Through this pathway (9Z,11E,13S)-13 hydroperoxy-9,11-octadecadienoic acid (13-HPOD) is reduced to corresponding 13 hydroxy acid (13-HOD), which is in turn dehydrogenated into ketodiene (9Z,11E,13S)-13-oxo-9,11-octadecadienoic acid (13-KOD). Dehydrogenation of 13-HOD into 13-KOD was not dependent on the presence of either NAD or NADP, but was strongly dependent on the presence of oxygen. Under anoxic conditions, 13-HOD dehydrogenation was blocked, but addition of 2,6-dichlorophenolindophenol restored it. Sulfite addition fully suppressed the aerobic dehydrogenation of 13 HOD. Hydrogen peroxide is a by-product formed by the enzyme along with 13-KOD. These data suggest that the ketodiene biosynthesis in H. tuberosus tubers is catalyzed by flavin dehydrogenase. (9S,10E,12Z)-9-Hydroxy-10,12-octadecadienoic acid (9-HOD) is dehydrogenated by this enzyme as effectively as 13-HOD, while alpha-ketol, (9Z)-12-oxo-13-hydroxy-9-octadecenoic acid, and ricinoleic acid did not act as substrates for dehydrogenase. The enzyme was soluble and possessed a pH optimum at pH 7.0-9.0. The only 13-HOD dehydrogenase known so far was detected in rat colon. However, unlike the H. tuberosus enzyme, the rat dehydrogenase is NAD-dependent. PMID- 15522818 TI - Group IIA secretory phospholipase A2 is a unique 12/15-lipoxygenase-regulated gene in cytokine-stimulated rat fibroblastic 3Y1 cells. AB - We have proposed previously that the expression of group IIA secretory phospholipase A(2) (sPLA(2)-IIA), an enzyme implicated in inflammation, is under the control of group IVA cytosolic phospholipase A(2) (cPLA(2)) and 12/15 lipoxygense (12/15-LOX) in cytokine-stimulated rat fibroblastic 3Y1 cells. Here, we show that the reduction of cytokine-stimulated sPLA(2)-IIA induction by the cPLA(2) inhibitor arachidonyl trifluoromethyl ketone (AACOCF(3)) is partially overcome by the addition of various lysophospholipids, such as lysophosphatidylcholine (LysoPC). Furthermore, this lysophospholipid effect was enhanced by further addition of 12/15-LOX products, such as 12(S)- or 15(S) hydroxyeicosatetraenoic acid (HETE) and 13(S)-hydroxyoctadecadienoic acid (HODE), thus substantiating the hypothesis that the expression of sPLA(2)-IIA is selectively regulated by lipid products of the cPLA(2)-12/15-LOX pathway. In an attempt to identify a set of 12/15-LOX-regulated genes, the cDNA subtraction technique, followed by Northern blotting, was performed to screen particular clones, the expression of which was suppressed by the LOX inhibitor nordihydroguaiaretic acid (NDGA). NDGA-sensitive clones identified thus far included sPLA(2)-IIA, cytoplasmic signaling intermediates, several oxygenases, extracellular matrices, secretory proteins, and other cellular proteins. Of these genes, however, only the expression of sPLA(2)-IIA and 14-3-3eta was enhanced by 12/15-LOX expression. Taken together, our data suggest that sPLA(2)-IIA represents a particular group of genes, the transcription of which is up regulated by 12/15-LOX metabolites. PMID- 15522819 TI - LXR activation and cholesterol efflux from a lipoprotein depot in vivo. AB - Activation of LXR in cultured cells results in enhancement of cholesterol efflux to apo Al. To study cholesterol efflux, in vivo cationized LDL was injected into the rectus femoris muscle of mice to create a lipoprotein depot. LXR ligand TO901317, 10 mg/kg, was given by gavage for 8 days, starting 4 days after injection of the lipoprotein. The rate of cholesterol efflux from the depot was compared in treated and control mice. Administration of the ligand resulted in a 70% increase in plasma cholesterol and 40% in phospholipids, but HDL-cholesterol and HDL-phospholipids increased by 43% and 24% only. Efflux of the injected cholesterol from the lipoprotein depot of treated mice was not enhanced but even somewhat delayed. This impairment was unexpected and its cause could be multifactorial. A plausible explanation seems that induced hypercholesterolemia, and a decrease in HDL-cholesterol to total cholesterol ratio, delayed the clearance. PMID- 15522820 TI - The ERG28-encoded protein, Erg28p, interacts with both the sterol C-4 demethylation enzyme complex as well as the late biosynthetic protein, the C-24 sterol methyltransferase (Erg6p). AB - In Saccharomyces cerevisiae, the C-24 sterol methyltransferase (Erg6p) converts zymosterol to fecosterol, an enzymatic step following C-4 demethylation of 4,4 dimethylzymosterol. Our previous study showed that an endoplasmic reticulum (ER) transmembrane protein, Erg28p, functions as a scaffold to tether the C-4 demethylation enzymatic complex (Erg25p-Erg26p-Erg27p) to the ER. To determine whether Erg28p also interacts with other ergosterol biosynthetic proteins, we compared protein levels of Erg3p, Erg6p, Erg7p, Erg11p and Erg25p in three pairs of erg28 and ERG28 strains. In erg28 strains, the Erg6p level in the ER fraction was decreased by about 50% relative to the wild-type strain, while ER protein levels of the four other ergosterol proteins showed no significant differences. Co-immunoprecipitation experiments, using an erg28 strain transformed with the epitope-tagged plasmid pERG28-HA and proteins detected with anti-HA and anti Erg6p antibodies, indicated that Erg6p and Erg28p reciprocally co immunoprecipitate. Further, the split ubiquitin yeast membrane two-hybrid system designed to detect protein interactions between membrane bound proteins also indicated an Erg28p-Erg6p interaction when pERG6-Cub was used as the bait and pERG28-NubG was used as the prey. We conclude that Erg28p may not only anchor the C-4 demethylation enzyme complex to the ER but also acts as a protein bridge to the Erg6p enzyme required for the next ergosterol biosynthetic step. PMID- 15522821 TI - Lipoprotein lipase-dependent binding and uptake of low density lipoproteins by THP-1 monocytes and macrophages: possible involvement of lipid rafts. AB - Lipoprotein lipase (LPL) is produced by cells in the artery wall and can mediate binding of lipoproteins to cell surface heparan sulfate proteoglycans (HSPG), resulting in endocytosis (the bridging function). Active, dimeric LPL may dissociate to inactive monomers, the main form found in plasma. We have studied binding/internalization of human low density lipoprotein (LDL), mediated by bovine LPL, using THP-1 monocytes and macrophages. Uptake of (125)I-LDL was similar in monocytes and macrophages and was not affected by the LDL-receptor family antagonist receptor-associated protein (RAP) or by the phagocytosis inhibitor cytochalasin D. In contrast, uptake depended on HSPG and on membrane cholesterol. Incubation in the presence of dexamethasone increased the endogenous production of LPL by the cells and also increased LPL-mediated binding of LDL to the cell surfaces. Monomeric LPL was bound to the cells mostly in a heparin resistant fashion. We conclude that the uptake of LDL mediated by LPL dimers is receptor-independent and involves cholesterol-enriched membrane areas (lipid rafts). Dimeric and monomeric LPL differ in their ability to mediate binding/uptake of LDL, probably due to different mechanisms for binding/internalization. PMID- 15522822 TI - Acyl chain-based molecular selectivity for HL60 cellular phosphatidylinositol and of phosphatidylcholine by phosphatidylinositol transfer protein alpha. AB - Mammalian phosphatidylinositol transfer protein alpha (PITP) is an intracellular lipid transporter with a binding site that can accommodate a single molecule of phosphatidylinositol (PI) or phosphatidylcholine (PC). Phospholipids are a heterogeneous population of molecular species that can be distinguished by their characteristic headgroups as well as their acyl chains at the sn-1 and sn-2 position. In this study, we have defined the acyl chain preference for PITPalpha when presented with a total population of cellular lipids. Recombinant PITPalpha loaded with bacterial lipid, phosphatidylglycerol (PG), was incubated with permeabilised HL60 cells, followed by recovery of PITPalpha by affinity chromatography. Lipids extracted from the PITPalpha were analysed by tandem electrospray ionisation mass spectrometry (ESI-MS) and showed total exchange of acquired bacterial lipids for HL60 cellular PI and PC. Detailed comparison of the molecular species composition of bound phospholipids with those in whole cells permitted the assessment of selectivity of acyl chain binding. For both phospholipid classes, progressive fractional enrichments in bound species possessing shorter acyl chains were apparent with a preference order: 16:1>16:0>18:1>18:0>20:4. A recapitulation of this specificity order was also seen from a dramatically altered range of molecular species present in HL60 cells enriched with arachidonate over many weeks of culture. We speculate that short chain, saturate-binding preferences under both conditions may reflect properties in vivo. This is consistent with target cell membranes actively remodelling newly delivered phospholipids after transport rather than relying on the transport of the specific molecular species conventionally found in mammalian membranes. PMID- 15522823 TI - Localization of various secretory phospholipase A2 enzymes in male reproductive organs. AB - Current evidence suggests the presence of transcripts for several secretory phospholipase A(2) (sPLA(2)) enzymes in male genital organs. In this study, we examined by immunohistochemistry the localization of group IIA, IIC, IID, IIE, IIF, V and X sPLA(2)s in male genital organs. In sPLA(2)-IIA-deficient C57BL/6 mouse testis, sPLA(2)-IIC, -IID, -IIE, -IIF, -V and -X were diversely expressed in spermatogenic cells within the seminiferous tubules. Immunoblotting revealed the presence of these sPLA(2)s in mouse spermatozoa. In addition, sPLA(2)-IIF, -V and -X were localized in the interstitial Leydig cells. The same set of sPLA(2)s was detected in a mouse cultured Leydig cell line, and adenovirus-mediated transfer of these sPLA(2)s into Leydig cells resulted in increased prostaglandin production. sPLA(2)-IIC, -IID, -IIE, -IIF, -V and -X were also detected diversely in the epithelium of the epididymis, vas deferens, seminal vesicles, and prostate. In a sPLA(2)-IIA-positive FVB strain, weak expression of sPLA(2)-IIA was detected in Leydig cells. Notable differences in the sPLA(2) expression profiles were found in the seminal vesicles and prostate between mice and humans. Taken together, individual sPLA(2)s exhibit distinct or partially overlapping localizations in male reproductive organs, suggesting both specific and redundant functions. PMID- 15522824 TI - Translocation of phospholipase A2 to membranes by oxidized LDL and hydroxyoctadecadienoic acid to contribute to cholesteryl ester formation. AB - We examined the mechanisms underlying the activation of group IVA cytosolic phospholipase A(2) (cPLA(2)alpha) contributing to the supply of fatty acids required for the formation of cholesteryl ester in oxidized low-density lipoprotein (oxLDL)-stimulated macrophages. The possible involvement of oxidized lipids was also examined. In [(3)H]arachidonic acid-labeled mouse peritoneal macrophages, oxLDL stimulated the release of arachidonic acid, which was suppressed by methyl arachidonyl fluorophosphonate (MAFP), a cPLA(2)alpha inhibitor. oxLDL induced an increase in PLA(2)alpha levels in the membrane fraction without affecting those in whole cells or the activity in the lysate. Among 13-hydroxyoctadecadienoic acid (13-HODE), 7-ketocholesterol, and 25 hydroxycholesterol, oxidized lipids present in oxLDL particles, only 13-HODE induced the release of arachidonic acid, which was also sensitive to MAFP. Under conditions where addition of Ca(2+) to the cell lysate induced an increase in cPLA(2)alpha protein in the membrane fraction, preincubation with 13-HODE facilitated the Ca(2+)-dependent translocation of cPLA(2)alpha. Furthermore, 13 HODE increased cholesteryl ester formation in the presence of [(3)H]cholesterol. These results suggest that 13-HODE mediates the oxLDL-induced activation of cPLA(2)alpha through an increase in cPLA(2)alpha protein in the membranes, thus contributing, in part, to the supply of fatty acids required for the esterification of cholesterol in macrophages. PMID- 15522825 TI - Contribution of lipid second messengers to the regulation of phosphatidylcholine synthesis during cell cycle re-entry. AB - During entry into the cell cycle a phosphatidylcholine (PC) metabolic cycle is activated. We have examined the hypothesis that PC synthesis during the G(0) to G(1) transition is controlled by one or more lipid products of PC turnover acting directly on the rate-limiting enzyme in the synthesis pathway, CTP: phosphocholine cytidylyltransferase (CCT). The acceleration of PC synthesis was two- to threefold during the first hour after addition of serum to quiescent IIC9 fibroblasts. The rate increased to approximately 15-fold above the basal rate during the second hour. The production of arachidonic acid, diacylglycerol (DAG), and phosphatidic acid (PA) preceded the second, rapid phase of PC synthesis. However, an increase in the cellular content of these lipid mediators was detected only for DAG. CCT activation and translocation to membranes accompanied the second phase of the PC synthesis acceleration. Bromoenol lactone (BEL), an inhibitor of calcium-independent phospholipase A(2) and PA phosphatase, blocked production of fatty acids and DAG, inhibited both phases of the PC synthesis response to serum, and reduced CCT activity and membrane affinity. The effect of BEL on PC synthesis was partially reversed by in situ generation of DAG via exogenous PC-specific phospholipase C to generate approximately 2-fold elevation in PC-derived DAG. Exogenous arachidonic acid also partially reversed the inhibition by BEL, but only at a concentration that generated a supra physiological cellular content of free fatty acid. 1-Butanol, which blocks PA production, had no effect on DAG generation, or on PC synthesis. We conclude that fatty acids and DAG could contribute to the initial slow phase of the PC synthesis response. DAG is the most likely lipid regulator of CCT activity and the rapid phase of PC synthesis. However, processes other than direct activation of CCT by lipid mediators likely contribute to the highly accelerated phase during entry into the cell cycle. PMID- 15522826 TI - The role of complex lipids in the synthesis of bioactive aldehydes of the marine diatom Skeletonema costatum. AB - Diatoms are unicellular plants broadly present in freshwater and marine ecosystems, where they play a primary role in sustaining the marine food chain. In the last 10 years, there has been accumulating evidence that diatoms may have deleterious effects on the hatching success of zooplankton crustaceans such as copepods, thus affecting dynamics of planktonic populations and limiting secondary production. At the molecular level, failure to hatch is ascribed to the presence of a family of inhibitory oxylipins, which we propose to collectively name polyunsaturated short-chain aldehydes (abbreviated here as PUSCAs). Here we describe the origin of PUSCAs produced by the marine diatom Skeletonema costatum via a lipoxygenase-mediated pathways involving non-esterified polyunsaturated fatty acids (PUFA). Experiments with complex lipids proved the pivotal role of chloroplast-derived glycolipids, especially monogalactosyldiacylglycerol (MGDG), in providing hexadecatrienoic acid (C16:3 omega-4), hexadecatetraenoic acid (C16:4 omega-1) and eicosapentaenoic acid (C20:5 omega-3) to the downstream process leading to 2E,4Z-octadienal (C8:2 omega-4), 2E,4Z,7-octatrienal (C8:3 omega-1) and 2E,4Z-heptadienal (C7:2 omega-3), respectively. Under physiological conditions, the hydrolytic process is associated to galactolipid hydrolyzing enzyme capable of removing fatty acids from both sn positions of glycerol. PMID- 15522827 TI - High-throughput quantification of phosphatidylcholine and sphingomyelin by electrospray ionization tandem mass spectrometry coupled with isotope correction algorithm. AB - The choline head group containing phosphatidylcholine (PC) and sphingomyelin (SPM) are major eukaryotic lipid components playing an important role in forming membrane microdomains and serve as precursor of signaling molecules. Both lipids can be monitored by positive ion mode electrospray tandem mass spectrometry using a parent ion scan of m/z 184. Although PC species appear at even m/z and SPM species at odd m/z, there may be a significant overlap of their isotopes. In order to separate PC and SPM species, an isotope correction algorithm was established, which utilizes calculated isotope percentages to correct the measured peak intensities for their isotopic overlap. We could demonstrate that this approach was applicable to correct the isotope overlap resulting from spiked PC and SPM species. Quantification was achieved by addition of different PC and SPM species prior to lipid extraction. The developed assay showed a precision, detection limit and robustness sufficient for routine analysis. Furthermore, an analysis time of only 1.3 min combined with automated data analysis using self programmed Excel Macros allows high-throughput analysis. In summary, this assay may be a valuable tool for detailed lipid analysis of PC and SPM species in a variety of sample materials. PMID- 15522828 TI - Differential effects of dietary fatty acids on rat liver alpha-amino-beta carboxymuconate-epsilon-semialdehyde decarboxylase activity and gene expression. AB - Hepatic alpha-amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD; formerly termed picolinic carboxylase) [EC4.1.1.45] plays a key role in regulating NAD biosynthesis and the generation of quinolinate (quinolinic acid) from tryptophan. Quinolinate is a potent endogenous excitotoxin of neuronal cells. We previously reported that ingestion of fatty acids by rats leads to a decrease in their hepatic ACMSD activity. However, the mechanism of this phenomenon is not clarified. We previously purified ACMSD and cloned cDNA encoding rat ACMSD. Therefore, in this study, we examined the differential effect of fatty acids on ACMSD mRNA expression by Northern blot. Moreover, we measured quinolinic acid concentration in rats fed on fatty acid. When diets containing 2% level of fatty acid were given to male Sprague-Dawley rats (4 weeks old) for 8 days, long-chain saturated fatty acids and oleic acid did not affect ACMSD mRNA expression in the liver. Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) strongly suppressed the liver ACMSD mRNA expression. In rats fed with high linoleic acid diet for 8 days, serum quinolinic acid was significantly increased as compared with the rats fed on a fatty acid-free diet under the condition of the approximately same calorie ingestion. These results suggest that the transcription level of ACMSD is modulated by polyunsaturated fatty acids, and suppressive potency of ACMSD mRNA is n-3 fatty acid family>linoleic acid (n-6 fatty acid)>saturated fatty acid. Moreover, this study provides the information that a high polyunsaturated fatty acid diet affects the production of quinolinic acid in serum by suppressing the ACMSD activity. PMID- 15522829 TI - Amyloid beta(1-42) and its beta(25-35) fragment induce activation and membrane translocation of cytosolic phospholipase A2 in bovine retina capillary pericytes. AB - We investigated changes in cytosolic phospholipase A(2) (cPLA(2)) and calcium independent PLA(2) (iPLA(2)) activities in bovine retina capillary pericytes after stimulation with 50 microM amyloid-beta (Abeta) (1-42) and its (25-35) fragment, over 24 h (mild, sublethal model of cell damage). In the presence of Abeta peptides, we found that cPLA(2) activity was increased and translocated from the cytosolic fraction to the membrane system, particularly in the nuclear region. Reversed-sequence Abeta(35-25) peptide did not stimulate or induce cPLA(2) translocation. Exposure to both Abeta peptides had no significant effect on cPLA(2) protein content as tested by Western immunoblot analysis. The addition of Abetas to quiescent pericytes was followed by phosphorylation of cPLA(2) and arachidonic acid release. Treatment with inhibitors (AACOCF(3), staurosporine and cycloheximide) resulted in a sharp decrease in basal and stimulated cPLA(2) activity. Inactivating effects of bromoenol lactone (BEL), inhibitor of iPLA(2), demonstrated that the stimulation of total PLA(2) activity by Abetas was mediated by both PLA(2) enzymes. Taken together with our previous observations that both Abeta peptides may induce hydrolysis of phosphatidylcholine, the present results provide evidence that this process is cooperatively mediated by cPLA(2) activation/translocation and iPLA(2) activation. The effect is very likely triggered by a mild prooxidant mechanism which was not able to divert the cell to degeneration. The data confirm the hypothesis that pericytes could be a target of potential vascular damage and reactivity during processes involving amyloid accumulation. PMID- 15522830 TI - The effect of dietary supplementation with linoleic acid to late gestation ewes on the fatty acid composition of maternal and fetal plasma and tissues and the synthetic capacity of the placenta for 2-series prostaglandins. AB - Linoleic acid (18:2n-6) is metabolised to arachidonic acid (20:4n-6), the precursor for 2-series prostaglandins (PGs). Increased consumption of 18:2n-6 during pregnancy may thus modify PG synthesis during labour. We have investigated whether increased 18:2n-6 composition during gestation altered the fatty acid consumption and PG synthesis of maternal and fetal tissues in the sheep. Ewes were fed a control diet or a diet providing 40% more 18:2n-6 from 96 days gestation. Half of each group received dexamethasone on day 136 to up-regulate the PG synthetic pathways promoting parturition. Maternal and fetal tissues were collected at 138 days. The 18:2n-6 diet significantly increased the 20:4n-6 content of maternal plasma, fetal plasma and allantochorion (51-81%) phosphatidylcholine, and fetal liver (40%) and maternal caruncular endometrium (57%) phosphatidylethanolamine. Increased 18:2n-6 intake increased production of PGF(2alpha) and PGE(2) in all placental tissues (maternal caruncular and intercaruncular endometrium and fetal allantochorion) by 23-98%, whereas dexamethasone increased it by 32-142%. This suggests that consumption of an 18:2n 6-enriched diet in late pregnancy enhanced placental PG production by increasing the supply of 20:4n-6. Variations in the extent to which the diet altered the polyunsaturated fatty acid (PUFA) content of the different tissues indicated complex interactions between nutrient availability and metabolic adaptation. PMID- 15522831 TI - Contribution of different biosynthetic pathways to species selectivity of aminoglycerophospholipids assembled into mitochondrial membranes of the yeast Saccharomyces cerevisiae. AB - In the yeast Saccharomyces cerevisiae, three pathways lead to the formation of cellular phosphatidylethanolamine (PtdEtn), namely the mitochondrial conversion of phosphatidylserine (PtdSer) to PtdEtn catalyzed by phosphatidylserine decarboxylase 1 (Psd1p), the equivalent reaction catalyzed by phosphatidylserine decarboxylase 2 (Psd2p) in the Golgi, and the CDP-ethanolamine branch of the so called Kennedy pathway which is located to the microsomal fraction. To investigate the contributions of these three pathways to the cellular pattern of PtdEtn species (fatty acid composition) we subjected lipids of wild-type and yeast mutant strains with distinct defects in the respective pathways to mass spectrometric analysis. We also analyzed species of PtdSer and phosphatidylcholine (PtdCho) of these strains because formation of the three aminoglycerophospholipids is linked through their biosynthetic route. We demonstrate that all three pathways involved in PtdEtn synthesis exhibit a preference for the formation of C34:2 and C32:2 species resulting in a high degree of unsaturation in total cellular PtdEtn. In PtdSer, the ratio of unsaturated to saturated fatty acids is much lower than in PtdEtn, suggesting a high species selectivity of PtdSer decarboxylases. Finally, PtdCho is characterized by its higher ratio of C16 to C18 fatty acids compared to PtdSer and PtdEtn. In contrast to biosynthetic steps, import of all three aminoglycerophospholipids into mitochondria of wild-type and mutant cells is not highly specific with respect to species transported. Thus, the species pattern of aminoglycerophospholipids in mitochondria is mainly the result of enzyme specificities, but not of translocation processes involved. Our results support a model that suggests equilibrium transport of aminoglycerophospholipids between mitochondria and microsomes based on membrane contact between the two compartments. PMID- 15522832 TI - Contribution of different pathways to the supply of phosphatidylethanolamine and phosphatidylcholine to mitochondrial membranes of the yeast Saccharomyces cerevisiae. AB - In the yeast, three biosynthetic pathways lead to the formation of phosphatidylethanolamine (PtdEtn): (i) decarboxylation of phosphatidylserine (PtdSer) by phosphatidylserine decarboxylase 1 (Psd1p) in mitochondria; (ii) decarboxylation of PtdSer by Psd2p in a Golgi/vacuolar compartment; and (iii) the CDP-ethanolamine (CDP-Etn) branch of the Kennedy pathway. The major phospholipid of the yeast, phosphatidylcholine (PtdCho), is formed either by methylation of PtdEtn or via the CDP-choline branch of the Kennedy pathway. To study the contribution of these pathways to the supply of PtdEtn and PtdCho to mitochondrial membranes, labeling experiments in vivo with [(3)H]serine and [(14)C]ethanolamine, or with [(3)H]serine and [(14)C]choline, respectively, and subsequent cell fractionation were performed with psd1Delta and psd2Delta mutants. As shown by comparison of the labeling patterns of the different strains, the major source of cellular and mitochondrial PtdEtn is Psd1p. PtdEtn formed by Psd2p or the CDP-Etn pathway, however, can be imported into mitochondria, although with moderate efficiency. In contrast to mitochondria, microsomal PtdEtn is mainly derived from the CDP-Etn pathway. PtdEtn formed by Psd2p is the preferred substrate for PtdCho synthesis. PtdCho derived from the different pathways appears to be supplied to subcellular membranes from a single PtdCho pool. Thus, the different pathways of PtdEtn biosynthesis play different roles in the assembly of PtdEtn into cellular membranes. PMID- 15522833 TI - Aging-related changes in ovarian hormones, their receptors, and neuroendocrine function. AB - Ovarian steroid hormones exert a broad range of effects on the body and brain. In the nervous system, estrogen and progesterone have crucial feedback actions on the hypothalamic neurons that drive the reproductive axis. In addition, hormones exert a variety of actions on other traditionally nonreproductive functions such as cognition, learning and memory, neuroprotection, mood and affective behavior, and locomotor activity. The actions of hormones on the hypothalamus are largely mediated by their nuclear hormone receptors, the two estrogen receptors, ERalpha and ERbeta, and the two progesterone receptor isoforms, PR-A and PR-B. Thus, changes in the circulating concentrations of estrogens and progestins during the life cycle can result in differential activation of their receptors. Furthermore, changes in the numbers, activity, and distribution of hypothalamic ERs and PRs can occur as a function of developmental age. The purpose of this article is to review the literature on the causes and consequences of alterations in steroid hormones, their neural receptors, and their interactions on reproductive senescence. We have also discussed several important experimental design considerations, focusing on rodent models in current use for understanding the mechanisms of menopause in women. PMID- 15522834 TI - DNA methylation, cancer susceptibility, and nutrient interactions. AB - DNA methylation is an important epigenetic mechanism of transcriptional control. DNA methylation plays an essential role in maintaining cellular function, and changes in methylation patterns may contribute to the development of cancer. Aberrant methylation of DNA (global hypomethylation accompanied by region specific hypermethylation) is frequently found in tumor cells. Global hypomethylation can result in chromosome instability, and hypermethylation has been associated with the inaction of tumor suppressor genes. Preclinical and clinical studies suggest that part of the cancer-protective effects associated with several bioactive food components may relate to DNA methylation patterns. Dietary factors that are involved in one-carbon metabolism provide the most compelling data for the interaction of nutrients and DNA methylation because they influence the supply of methyl groups, and therefore the biochemical pathways of methylation processes. These nutrients include folate, vitamin B(12), vitamin B(6), methionine, and choline. However, looking at individual nutrients may be too simplistic. Dietary methyl (folate, choline, and methionine) deficiency in combination causes decreased tissue S-adeno-sylmethionine, global DNA hypomethylation, hepatic steatosis, cirrhosis, and ultimately hepatic tumorigenesis in rodents in the absence of carcinogen treatment. Other dietary components such as vitamin B(12), alcohol, and selenium may modify the response to inadequate dietary folate. PMID- 15522835 TI - Unraveling the molecular details involved in the intimate link between the immune and neuroendocrine systems. AB - During systemic infections, the immune system can signal the brain and act on different neuronal circuits via soluble molecules, such as proinflammatory cytokines, that act on the cells forming the blood-brain barrier and the circumventricular organs. These activated cells release prostaglandin of the E(2) type (PGE(2)), which is the endogenous ligand that triggers the pathways involved in the control of autonomic functions necessary to restore homeostasis and provide inhibitory feedback to innate immunity. Among these neurophysiological functions, activation of the circuits that control the plasma release of glucocorticoids is probably the most critical to the survival of the host in the presence of pathogens. This review revisits this issue and describes in depth the molecular details (including the emerging role of Toll-like receptors during inflammation) underlying the influence of circulating inflammatory molecules on the cerebral tissue, focusing on their contribution in the synthesis and action PGE(2) in the brain. We also provide an innovative view supporting the concept of "fast and delayed response" involving the same ligands but different groups of cells, signal transduction pathways, and target genes. PMID- 15522836 TI - Ghrelin expression and actions: a novel peptide for an old cell type of the diffuse endocrine system. AB - Ghrelin is a gastric peptide involved in food intake control and growth hormone release. Its cell localization has been defined in distinct ghrelin cells of the gastric mucosa in humans and other mammals. Ghrelin production was also described in a number of other sites of the diffuse endocrine system, including the pituitary, thyroid, lung, pancreas, adrenal gland, and intestine. In addition, ghrelin cells were identified early during fetal life and in the placenta and gonads. Finally, endocrine growths and tumors of the diffuse endocrine system may present ghrelin-producing cells, and in a few cases high levels of circulating ghrelin were reported. Besides its well-defined orexigenic role, ghrelin is likely to exert a local paracrine role similar to other brain-gut axis hormones. This review aims to summarize recent data on ghrelin cell distribution in the diffuse endocrine system and discuss local and general ghrelin function during development, adulthood, and endocrine tumor development. PMID- 15522837 TI - Dietary corn oil promotes colon cancer by inhibiting mitochondria-dependent apoptosis in azoxymethane-treated rats. AB - How dietary corn oil is involved in colon carcinogenesis and cancer development is poorly understood. The aim of this study was to investigate whether long-term dietary corn oil promotes colon cancer by inhibiting the tumor suppressor gene p53-mediated mitochondria-dependent apoptosis in azoxymethane (AOM)-treated rats. Male Sprague-Dawley rats were injected with AOM or with saline and fed on a basal diet or basal diet supplemented with 10% corn oil for 48 weeks. Colonic aberrant crypt foci (ACF) and tumors, including adenomas and carcinomas, were examined. Colonic apoptosis and cell proliferation were evaluated. Wild type (wt) p53 was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. In addition, Bcl-2, Bcl-xL, Bax, and Bak localized in the mitochondria were detected. Long-term dietary corn oil increased ACF in AOM treated rats at 12 weeks and promoted colon cancer invasion at 48 weeks. Cancer invasion was not observed in the AOM-treated rats without dietary corn oil, although colon adenomas and cancers were detected. Apoptosis was decreased and cell proliferation was increased in the AOM-treated rats with dietary corn oil, compared with the AOM-treated rats with dietary basal diet. In these rats, mitochondrial wt p53 was significantly inhibited through decreased mitochondrial localization of wt p53 and increased cytosolic p53, resulting in the upregulation of Bcl-2 and Bcl-xL and the downregulation of Bak in the mitochondria. Results suggest that long-term dietary corn oil promotes AOM-induced colon cancer development partly by inhibiting the tumor suppressor gene p53-mediated mitochondria-dependent apoptosis. PMID- 15522838 TI - Extracellular cyclic AMP and adenosine appearance in adipose tissue of Sus scrofa: effects of exercise. AB - Cyclic AMP (cAMP) appears extracellularly in a variety of tissues including brain, liver, and kidney; whether it appears in adipose tissue and responds to physiological perturbation is unknown. The purpose of this study was to examine adipose tissue extracellular cAMP appearance and metabolism in situ and in vitro in physiologically challenged animals. Littermate swine were either sedentary or exercise trained on a treadmill for 3 months and subjected to acute exercise on experiment day. In situ, microdialysis probes in subcutaneous back fat were perfused before, during, and after animals performed 20 mins of acute exercise, and dialysate was analyzed for cAMP and adenosine. In vitro, isolated adipocytes were hormonally stimulated to provoke cAMP synthesis and efflux, and plasma membrane phosphodiesterase and 5'-nucleotidase activities were measured. Extracellular cAMP and adenosine levels in adipose tissue of sedentary swine averaged 5.2 +/- 1.7 and 863 +/- 278 nM, respectively. Exercise training tended to increase extracellular cAMP (11.3 +/- 1.7 nM) and reduce extracellular adenosine (438 +/- 303 nM), although neither change was statistically significant. Acute exercise caused a significant 3-fold and 16-fold increase in extracellular cAMP and adenosine, respectively, compared to rest. These changes occurred despite a 2- to 3-fold increase in adipose tissue blood flow during acute exercise. In vitro, cAMP efflux from exercise-trained swine was 42% greater than that from adipocytes of sedentary swine, yet adipocyte plasma membranes from exercise-trained and sedentary swine did not differ in maximal phosphodiesterase and 5'-nucleotidase activities. We conclude that cAMP appears extracellularly in swine adipose tissue and that the levels of extracellular cAMP and adenosine in intact swine adipose tissue are influenced by both acute and chronic exercise. The subsequent impact of the changes in these biochemicals on local cellular metabolism and growth remains to be determined. PMID- 15522839 TI - Leptin acts peripherally to limit meal-induced increases in plasma insulin concentrations in mice: a brief communication. AB - Leptin inhibits food intake and lowers plasma insulin concentrations. This study was designed to determine whether leptin acts independent of food-intake regulation to affect meal-induced increases in plasma insulin concentrations. Leptin-deficient, Lep(ob)/Lep(ob) mice were administered 1 microg leptin intracerebroventricularly (ICV) or intraperitoneally. Food intake and plasma insulin concentrations of mice administered leptin ICV before a meal were lower, as expected, than were intakes and plasma insulin concentrations of mice administered vehicle ICV. However when food intake was controlled, meal-induced increases in plasma insulin were unaffected by ICV administration of leptin. Intraperitoneal administration of 1 microg leptin before a meal lowered meal induced increases in plasma insulin concentrations without influencing the size of the meal. We conclude that plasma leptin concentrations can affect meal induced insulin secretion independent of the central nervous system actions of leptin associated with food-intake regulation. PMID- 15522840 TI - Aminoguanidine prevents fructose-induced arterial stiffening in Wistar rats: aortic impedance analysis. AB - Fructose has been reported as a potent agent in forming advanced glycation end products (AGEs) and, thus, may play a significant role in the pathogenesis of diabetic complications. Herein, we determined the effects of aminoguanidine (AG), an inhibitor of AGEs, on the mechanical properties of the arterial system in fructose-fed (FF) rats, using aortic impedance analysis. Rats at 2 months were given 10% fructose in drinking water for 2 weeks and compared with untreated age matched controls. Meanwhile, FF rats were treated for 2 weeks with AG (daily peritoneal injections of 50 mg kg(-1)) and compared with the untreated FF group. Neither fructose nor AG affects body weight, blood glucose level, and basal heart rate. In comparison with controls, FF rats showed a decrease in cardiac output in the absence of any significant changes in mean aortic pressure, having increased total peripheral resistance (R(p)), at 51.1 +/- 2.9 versus 66.2 +/- 1.9 mm Hg sec ml(-1) (P < 0.05). Fructose also contributed to an increase in aortic characteristic impedance (Z(c)), from 1.528 +/- 0.094 to 1.933 +/- 0.084 mm Hg sec ml (-1) (P < 0.05) and a decrease in wave transit time (tau), from 22.6 +/- 0.6 to 19.2 +/- 0.7 msec (P < 0.05). The elevated Z(c) and the reduced tau suggest that fructose may cause a detriment to the aortic distensibility in animals. After exposure to AG, FF rats exhibited a significant improvement in physical properties of the resistance vessels, as evidenced by the reduction of 21.3% in R(p). Meanwhile, AG retarded the fructose-induced decline in aortic distensibility, as reflected in the decrease of 16.0% in Z(c) (P < 0.05) and the increase of 18.1% in tau (P < 0.05). By contrast, AG exerted no effects on the mechanical properties of Windkessel vessels, as well as resistance vessels, in normal diet controls. We conclude that AG may prevent the fructose-derived changes in arterial stiffening, possibly through inhibition of the fructose derived advanced glycation end product formation in Wistar rats. PMID- 15522841 TI - Low-density lipoprotein inhibits secretion of phospholipid transfer protein in human trophoblastic BeWo cells. AB - Human plasma phospholipid transfer protein (PLTP) plays an important role in lipoprotein metabolism. In this study, we investigated the effects of lipoproteins on the secretion of PLTP in cultured BeWo choriocarcinoma cells. Low density lipoproteins (LDLs) decreased PLTP secretion in a dose- and time dependent manner, whereas very low density lipoproteins and high-density lipoproteins (HDLs) had little effect. LDL suppression of PLTP secretion was not altered by the inhibition of both LDL receptor and LDL receptor-related protein with receptor-associated protein. Mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor, U0126, could abolish the LDL-mediated inhibition of PLTP secretion. Furthermore, LDL, but not HDL, could stimulate the expression of MAPK phosphatase-1 (MKP-1) in BeWo cells that resulted in the inactivation of p44/p42 extracellular signal-regulated kinase (ERK) 1 and 2, the family members of MAPKs. These results support the conclusion that LDL-mediated suppression of PLTP secretion in BeWo cells is through a LDL receptor-independent MAPK signaling pathway. PMID- 15522842 TI - 9HODE stimulates cell proliferation and extracellular matrix synthesis in human mesangial cells via PPARgamma. AB - Plasma oxidized low-density lipoprotein (OX-LDL) levels are elevated in patients with renal diseases, including diabetic nephropathy. We examined effects of OX LDL on cell proliferation and extracellular matrix (ECM) production by using normal human mesangial cells. Furthermore, we examined possible involvement of peroxisome proliferator-activated receptor gamma (PPARgamma). Mesangial cell proliferation with OX-LDL, 9-hydroxy-10,12-octadecadienoic acid (9HODE), and 13 hydroxy-9,11-octadecadienoic acid (13HODE), the major components of OX-LDL, were determined by 5-bromo-2'-deoxyuridine (BrdU) or 3-(4,5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide (MTT) incorporation. The effect of OX-LDL on mesangial cell proliferation with PD98059 pretreatment was determined by BrdU incorporation. Type IV collagen, fibronectin, and PPARgamma expression with OX LDL or 9HODE or 13HODE was determined by Western blotting. Type IV collagen expression with antisense oligonucleotide against PPARgamma pretreatment was also determined by Western blotting. The effect of PD98059 pretreatment on PPARgamma expression was determined by Western blotting. In mesangial cells exposed to isolated OX-LDL from human plasma, BrdU incorporation was increased, and this increase was deleted by PD98059. Type IV collagen expression was significantly increased by OX-LDL. 9HODE and 13HODE increased BrdU and MTT incorporation into mesangial cells and also increased expressions of Type IV collagen and fibronection, the major components of ECM. PPARgamma expression in mesangial cells was stimulated by 9HODE. The reduction of PPARgamma synthesis by pretreatment of antisense oligonucleotide against PPARgamma remarkably attenuated Type IV collagen synthesis induced by 9HODE. PPARgamma expression induced by 9HODE was also reduced by PD98059 pretreatment. These findings demonstrate that 9HODE, the major component of OX-LDL, stimulates cell proliferation and ECM production of human mesangial cells. In addition, the stimulatory effects are, at least in part, mediated by PPARgamma, which may exist in downstream of ERK1/2 pathway. PMID- 15522843 TI - Functional piglet model for the clinical syndrome and postmortem findings induced by staphylococcal enterotoxin B. AB - Staphylococcal enterotoxin (SE) B causes serious gastrointestinal illness, and intoxication with this exotoxin can lead to lethal toxic shock syndrome. In order to overcome significant shortcomings of current rodent and nonhuman primate models, we developed a piglet model of lethal SEB intoxication. Fourteen-day-old Yorkshire piglets were given intravenous SEB, observed clinically, and sacrificed at 4, 6, 24, 48, 72, or 96 hrs posttreatment. Clinical signs were biphasic with pyrexia, vomiting, and diarrhea within 4 hrs, followed by terminal hypotension and shock by 96 hrs. Mild lymphoid lesions were identified as early as 24 hrs, with severe lymphadenopathy, splenomegaly, and prominent Peyer's patches found by 72 hrs. Widespread edema-most prominent in the mesentery, between loops of spiral colon, and in retroperitoneal connective tissue-was found in animals at 72 hrs. Additional histologic changes included perivascular aggregates of large lymphocytes variably present in the lung and brain, circulating lymphoblasts, and lymphocytic portal hepatitis. Preliminary molecular investigation using gene array has uncovered several gene profile changes that may have implications in the pathophysiology leading to irreversible shock. Five genes were selected for further study, and all showed increased mRNA levels subsequent to SEB exposure. The use of this piglet model will continue to elucidate the pathogenesis of SEB intoxication and facilitate the testing of new therapeutic regimens that may better correlate with human lesions. PMID- 15522844 TI - Apoptotic and necrotic mechanisms of stress-induced human lens epithelial cell death. AB - Exposure to ultraviolet radiation (UVR) and reactive oxygen species (ROS) can damage the human lens and contribute to cataract formation. Recent evidence suggests that apoptosis in lens epithelial cells (LEC) is an initiating event in noncongenital cataract formation in humans and animals. The present study examines the cellular and molecular mechanisms by which environmental (ultraviolet B [UVB]) and chemical (hydrogen peroxide [H(2)O(2)], t-butyl hydroperoxide [TBHP]) stress induces cell death in an SV-40 immortalized human lens epithelial (HLE) cell line. Treatment of HLE cells with UVB, H(2)O(2), and TBHP significantly decreased cell density with LD50 values of 350 J/m(2), 500 muM, and 200 muM, respectively. Cellular morphology, DNA fragmentation, and annexin/propidium iodide staining consistent with apoptosis was observed only in UVB-treated cells, whereas lactate dehydrogenase (LDH) release was significantly higher in H(2)0(2)- and TBHP-treated cells. In addition, activation of apoptotic stress-signaling proteins, including c-Jun NH2-terminal kinase (JNK), caspase-3, and DNA fragmentation factor 45 (DFF45) was observed only in UVB-treated cells. Inhibition of JNK activity increased UVB-induced cell death, suggesting that this pathway may serve a prosurvival role in HLE cells. These findings suggest UVB predominantly induces apoptosis in HLE cells, whereas H(2)O(2) and TBHP induce necrosis. PMID- 15522845 TI - Complementary DNA microarray analysis in acute lung injury induced by lipopolysaccharide and diesel exhaust particles. AB - We have recently shown that diesel exhaust particles (DEP) synergistically enhance acute lung injury related to lipopoly-saccharide (LPS) in mice. The present study used cDNA microarray to elucidate the effects of DEP on the global pattern of LPS-related gene expression in the murine lung. The number of genes upregulated >/=2-fold as compared with their expression levels in the vehicle group was greater in the LPS group than in other groups, but treatment with DEP and LPS dramatically increased the number of the genes upregulated >/=6-fold. In particular, gene expression of metallothionein-1 and -2, S100 calcium-binding protein A9, lipocalin 2, and small inducible cytokine B family member 10 was higher by >/=20-fold in the DEP + LPS group than in the vehicle group. These results were concomitant with those obtained by real-time reverse transcription polymerase chain reaction analysis in the overall trend. Our findings suggest that intense, focused expression of genes such as S100 calcium-binding protein A9, lipocalin 2, and small inducible cytokine B family member 10 relates to the synergistic aggravation of acute lung injury by LPS and DEP rather than weak, broad expression of various genes by exposure of LPS alone. PMID- 15522846 TI - S-RNase and SLF determine S-haplotype-specific pollen recognition and rejection. PMID- 15522847 TI - A puromycin-sensitive aminopeptidase is essential for meiosis in Arabidopsis thaliana. AB - Puromycin-sensitive aminopeptidases (PSAs) participate in a variety of proteolytic events essential for cell growth and viability, and in fertility in a broad range of organisms. We have identified and characterized an Arabidopsis thaliana mutant (mpa1) from a pool of T-DNA tagged lines that lacks PSA activity. This line exhibits reduced fertility, producing shorter siliques (fruits) bearing a lower number of seeds compared with wild-type plants. Cytogenetic characterization of meiosis in the mutant line reveals that both male and female meiosis are defective. In mpa1, early prophase I appears normal, but after pachytene most of the homologous chromosomes are desynaptic, thus, by metaphase I a high level of univalence is observed subsequently leading to abnormal chromosome segregation. Wild-type plants treated with specific inhibitors of PSA show a very similar desynaptic phenotype to that of the mutant line. A fluorescent PSA-specific bioprobe, DAMPAQ-22, reveals that the protein is maximally expressed in wild-type meiocytes during prophase I and is absent in mpa1. Immunolocalization of meiotic proteins showed that the meiotic recombination pathway is disrupted in mpa1. Chromosome pairing and early recombination appears normal, but progression to later stages of recombination and complete synapsis of homologous chromosomes are blocked. PMID- 15522848 TI - Gain and loss of fruit flavor compounds produced by wild and cultivated strawberry species. AB - The blends of flavor compounds produced by fruits serve as biological perfumes used to attract living creatures, including humans. They include hundreds of metabolites and vary in their characteristic fruit flavor composition. The molecular mechanisms by which fruit flavor and aroma compounds are gained and lost during evolution and domestication are largely unknown. Here, we report on processes that may have been responsible for the evolution of diversity in strawberry (Fragaria spp) fruit flavor components. Whereas the terpenoid profile of cultivated strawberry species is dominated by the monoterpene linalool and the sesquiterpene nerolidol, fruit of wild strawberry species emit mainly olefinic monoterpenes and myrtenyl acetate, which are not found in the cultivated species. We used cDNA microarray analysis to identify the F. ananassa Nerolidol Synthase1 (FaNES1) gene in cultivated strawberry and showed that the recombinant FaNES1 enzyme produced in Escherichia coli cells is capable of generating both linalool and nerolidol when supplied with geranyl diphosphate (GPP) or farnesyl diphosphate (FPP), respectively. Characterization of additional genes that are very similar to FaNES1 from both the wild and cultivated strawberry species (FaNES2 and F. vesca NES1) showed that only FaNES1 is exclusively present and highly expressed in the fruit of cultivated (octaploid) varieties. It encodes a protein truncated at its N terminus. Green fluorescent protein localization experiments suggest that a change in subcellular localization led to the FaNES1 enzyme encountering both GPP and FPP, allowing it to produce linalool and nerolidol. Conversely, an insertional mutation affected the expression of a terpene synthase gene that differs from that in the cultivated species (termed F. ananassa Pinene Synthase). It encodes an enzyme capable of catalyzing the biosynthesis of the typical wild species monoterpenes, such as alpha-pinene and beta-myrcene, and caused the loss of these compounds in the cultivated strawberries. The loss of alpha-pinene also further influenced the fruit flavor profile because it was no longer available as a substrate for the production of the downstream compounds myrtenol and myrtenyl acetate. This phenomenon was demonstrated by cloning and characterizing a cytochrome P450 gene (Pinene Hydroxylase) that encodes the enzyme catalyzing the C10 hydroxylation of alpha pinene to myrtenol. The findings shed light on the molecular evolutionary mechanisms resulting in different flavor profiles that are eventually selected for in domesticated species. PMID- 15522849 TI - Guns in the home and risk of a violent death in the home: findings from a national study. AB - Data from a US mortality follow-back survey were analyzed to determine whether having a firearm in the home increases the risk of a violent death in the home and whether risk varies by storage practice, type of gun, or number of guns in the home. Those persons with guns in the home were at greater risk than those without guns in the home of dying from a homicide in the home (adjusted odds ratio = 1.9, 95% confidence interval: 1.1, 3.4). They were also at greater risk of dying from a firearm homicide, but risk varied by age and whether the person was living with others at the time of death. The risk of dying from a suicide in the home was greater for males in homes with guns than for males without guns in the home (adjusted odds ratio = 10.4, 95% confidence interval: 5.8, 18.9). Persons with guns in the home were also more likely to have died from suicide committed with a firearm than from one committed by using a different method (adjusted odds ratio = 31.1, 95% confidence interval: 19.5, 49.6). Results show that regardless of storage practice, type of gun, or number of firearms in the home, having a gun in the home was associated with an increased risk of firearm homicide and firearm suicide in the home. PMID- 15522850 TI - Incidence and risk factors for concussion in high school athletes, North Carolina, 1996-1999. AB - A prospective cohort study was used to quantify risk factors for sports concussions. Analysis was based on a stratified cluster sample of North Carolina high school athletes followed during 1996-1999. Clustering was by school and sport, and the sample included 15,802 athletes with 1-8 seasons of follow-up per athlete. Concussion rates were estimated for 12 sports, and risk factors were quantified using generalized Poisson regression. Concussion rates ranged from 9.36 (95% confidence interval: 1.93, 16.80) per 100,000 athlete-exposures in cheerleading to 33.09 (95% confidence interval: 24.74, 41.44) per 100,000 athlete exposures in football, where "athlete-exposure" is one athlete participating in one practice or game. The overall rate of concussion was 17.15 (95% confidence interval: 13.30, 21.00) per 100,000 athlete-exposures. Cheerleading was the only sport for which the practice rate was greater than the game rate. Almost two thirds of cheerleading concussions involved two-level pyramids. Concussion rates were elevated for athletes with a history of concussion, and they increased with the increasing level of body contact permitted in the sport. After adjustment for sport, body mass index, and year in school, history of concussion(s) remained a moderately strong risk factor for concussion (rate ratio = 2.28, 95% confidence interval: 1.24, 4.19). The fact that concussion history is an important predictor of concussion incidence, even in this young population, underscores the importance of primary prevention efforts, timely identification, and careful clinical management of these injuries. PMID- 15522851 TI - A case-control study of risk factors for fibrocystic breast conditions: Shanghai Nutrition and Breast Disease Study, China, 1995-2000. AB - This study was conducted to identify reproductive and dietary factors associated with benign proliferative mammary epithelial cell changes. Subjects were women enrolled in a randomized trial of breast self-examination in Shanghai, China. Women who developed fibrocystic breast conditions classified as nonproliferative (175 women), proliferative (181 women), or proliferative with atypia (33 women) between 1995 and 2000 and 1,070 unaffected trial participants were administered general risk factor and food frequency questionnaires. Conditional logistic regression was used to estimate adjusted odds ratios and 95% confidence intervals. High parity and consumption of fresh fruits and vegetables were more strongly associated with a reduced risk of proliferative and atypical lesions than with nonproliferative conditions. For the fourth quartile of consumption versus the first, odds ratios for lesions diagnosed as nonproliferative, proliferative, and proliferative with atypia were 0.4 (95% confidence interval (CI): 0.2, 0.7), 0.2 (95% CI: 0.1, 0.4), and 0.1 (95% CI: 0.03, 0.5), respectively, for fruit intake and 0.6 (95% CI: 0.3, 1.1), 0.4 (95% CI: 0.2, 0.7), and 0.1 (95% CI: 0.1, 0.9), respectively, for vegetable intake. Reduced but nonsignificant risks in relation to soy products were observed for proliferative and atypical lesions. No single nutrient or botanical family was appreciably more strongly associated with proliferative conditions than with nonproliferative conditions, after results were controlled for total fruit and vegetable consumption. A diet rich in fruits and vegetables may reduce cellular proliferation in the mammary epithelium; this is one mechanism by which such a diet could reduce risk of breast cancer. PMID- 15522852 TI - A prospective cohort study of nut consumption and the risk of gallstone disease in men. AB - Gallstone disease is a major source of morbidity in Western countries. Nuts are rich in several compounds that may be protective against gallstones. The objective of the study was to examine the relation between nut consumption and gallstone disease in the Health Professionals Follow-up Study. The consumption of nuts was assessed starting in 1986 as part of a 131-item semiquantitative food frequency questionnaire. The main outcome measure was newly diagnosed symptomatic gallstones. During 457,305 person-years of follow-up, 1,833 participants reported gallstone disease. After adjustment for age and other known or suspected risk factors, men consuming 5 or more units of nuts per week (frequent consumption) had a significantly lower risk of gallstone disease (relative risk = 0.70, 95% confidence interval: 0.60, 0.86; p(trend) < 0.001) than did men who never ate or who ate less than 1 unit per month (rare consumption) (1 unit = 1 ounce (0.028 kg) of nuts). Further adjustment for fat consumption (saturated fat, trans-fat, polyunsaturated fat, and monounsaturated fat) did not materially alter the relation. In analyses examining consumption of peanuts and other nuts separately, both were significantly associated with a lower risk of gallstone disease in the age-adjusted and multivariate models. Our findings suggest that frequent nut consumption is associated with a reduced risk of gallstone disease in men. PMID- 15522853 TI - Body mass index in relation to adult asthma among 135,000 Norwegian men and women. AB - The authors estimated the association between asthma and body mass index in a 1963-2002 study of 135,000 Norwegians aged 14-60 years who were followed on average for 21 years. Cox proportional hazards regression models were fitted to estimate the relative risk of asthma adjusting for smoking, education, and physical activity. Compared with persons with a body mass index (weight (kg)/height (m)(2)) of less than 25, overweight (body mass index: 25-29) men and women had relative risks of asthma of 1.27 (95% confidence interval (CI): 1.13, 1.43) and 1.30 (95% CI: 1.17, 1.45), respectively, while obese (body mass index: >/=30) men and women had relative risks of 1.78 (95% CI: 1.35, 2.34) and 1.99 (95% CI: 1.67, 2.37), respectively. Stratified analyses revealed a similar association between body mass index and asthma for never smokers, ever smokers, persons with less than or equal to 12 years of education, and persons with more than 12 years of education. Analyses including all the covariates gave results similar to those not adjusting for these factors. The risk of asthma increased steadily with body mass index, from a body mass index of 20 in men and of 22 in women. In men, the risk of asthma increased by 10% with each unit of increased body mass index between 25 and 30. The similar value for women was 7%. Overweight or obese persons reported asthma more often than did thinner persons after adjustment for smoking, education, and physical activity. PMID- 15522854 TI - Coffee consumption, gender, and Parkinson's disease mortality in the cancer prevention study II cohort: the modifying effects of estrogen. AB - Caffeine consumption is associated with a reduced risk of Parkinson's disease in men but not in women. This gender difference may be due to an interaction between caffeine and use of postmenopausal estrogens. The authors prospectively assessed the relation between coffee consumption and Parkinson's disease mortality among participants in the Cancer Prevention Study II, a cohort of over 1 million people enrolled in 1982. Causes of deaths were ascertained through death certificates from January 1, 1989, through 1998. Parkinson's disease was listed as a cause of death in 909 men and 340 women. After adjustment for age, smoking, and alcohol intake, coffee consumption was inversely associated with Parkinson's disease mortality in men (p(trend) = 0.01) but not in women (p = 0.6). In women, this association was dependent on postmenopausal estrogen use; the relative risk for women drinking 4 or more cups (600 ml) of coffee per day compared with nondrinkers was 0.47 (95% confidence interval: 0.27, 0.80; p = 0.006) among never users and 1.31 (95% confidence interval: 0.75, 2.30; p = 0.34) among users. These results suggest that caffeine reduces the risk of Parkinson's disease but that this hypothetical beneficial effect may be prevented by use of estrogen replacement therapy. PMID- 15522855 TI - The epidemiology of acute pyelonephritis in South Korea, 1997-1999. AB - Acute pyelonephritis causes significant morbidity, tends to recur, and can be fatal; however, little is known regarding its epidemiology. In this paper, the authors describe the epidemiology of acute pyelonephritis in South Korea by using nationwide heath insurance claims data from 1997 to 1999. The National Health Insurance System of South Korea covers almost the entire population (99%). The overall average annual incidence rate of pyelonephritis in 1997-1999 was 35.7 per 10,000 population (male, 12.6; female, 59.0). Approximately one of every seven patients was hospitalized (incidence per 10,000: inpatients, 5.5; outpatients, 30.1). Incidence varied with age and was higher in the summer season. Following an initial episode, the risk of a second episode within 12 months was 9.2% for females and 5.7% for males; by contrast, the risk of a fifth episode within a year following a fourth episode was 50.0% for females and 53.0% for males. Female sex (hazard ratio = 1.89, 95% confidence interval: 1.60, 2.23), advancing age, outpatient treatments (hazard ratio = 1.35, 95% confidence interval: 1.14, 1.60), and medical aid (hazard ratio = 1.23, 95% confidence interval: 1.08, 1.40) increased the risk of any recurrence. Pyelonephritis has a clear seasonal pattern and high rate of recurrence. The incidence of hospitalization for pyelonephritis in South Korea is similar to that in the United States and Canada. PMID- 15522856 TI - Estimating long-term trends in the incidence and prevalence of opiate use/injecting drug use and the number of former users: back-calculation methods and opiate overdose deaths. AB - The authors explored an age-specific back-calculation approach to estimating long term trends in the incidence and prevalence of opiate use/injecting drug use (IDU) in England for 1968-2000. The incidence of opiate use/IDU was estimated by combining information on the observed opiate overdose deaths of persons aged 15 44 years with knowledge on the distribution of the time between starting opiate use/IDU and death by overdose (incubation time distribution). The resulting incidence, together with the incubation time distribution, other drug-related mortality, and the general age-specific mortality rate, was then used to estimate the prevalence of current and former users. Provisional estimates suggested two major increases in incidence in the late 1970s and early 1990s, with models including information on age at death suggesting a recent decline since 1997 and that prevalence of opiate use/IDU increased substantially in the 1990s. Results were crucially dependent on assumptions about key parameters of the back calculation framework. In theory, the approach is a valuable addition to the portfolio of indirect methods for estimating incidence and prevalence of dependent opiate use/IDU. In practice, its full potential will be realized only once better information on the process of stopping opiate use/IDU becomes available and more precise estimates of current and historical overdose mortality are obtained. PMID- 15522857 TI - Fish intake, marine omega-3 fatty acids, and mortality in a cohort of postmenopausal women. AB - Intake of fish or omega-3 fatty acids may decrease risk of total and coronary heart disease death, but evidence from low-risk populations is less convincing. The authors assessed intake by using a food frequency questionnaire at baseline in a cohort of Iowa women aged 55-69 years. Among women initially free of heart disease and cancer (4,653 deaths over 442,965 person-years), there was an inverse age- and energy-adjusted association between total mortality and fish intake, with a relative risk of 0.82 (95% confidence interval: 0.74, 0.91) for the highest versus lowest quintile. Age- and energy-adjusted associations also were inverse (p for trend < 0.05), although not entirely monotonic, for cardiovascular, coronary heart disease, and cancer mortality. Adjustment for multiple other risk factors attenuated all associations to statistically nonsignificant levels. Estimated marine omega-3 fatty acid intake also was not associated with total or cause-specific mortality. In comparison, plant-derived alpha-linolenic acid was inversely associated with mortality after multivariable adjustment. Intake of neither fish nor marine omega-3 fatty acids was associated with breast cancer incidence. These findings do not argue against recommending fish as part of a healthy diet, as other evidence suggests benefit. Nevertheless, the authors of this 1986-2000 study could not verify that fish and marine omega-3 fatty acid intake had independent health benefits in these postmenopausal women. PMID- 15522858 TI - Dietary fat and fatty acids and risk of colorectal cancer in women. AB - The authors examined the association of intakes of different types of fat and fatty acids with risk of colorectal cancer using data from the Women's Health Study, a randomized trial of low-dose aspirin and vitamin E carried out among 39,876 healthy US women aged >/=45 years. Among the 37,547 women eligible for the present study, 202 developed colorectal cancer during an average follow-up period of 8.7 years (1993-2003). Intakes of dietary fat and its food sources were assessed at baseline by food frequency questionnaire. Cox proportional hazards regression was used to estimate relative risks and 95% confidence intervals. Total fat intake was not related to colorectal cancer risk, nor were intakes of the different types of fat and major fatty acids. However, the authors observed a positive association between intake of fried foods away from home and colorectal cancer risk (highest quintile vs. lowest: relative risk = 1.86, 95% confidence interval: 1.09, 3.16; p for trend = 0.01). These prospective cohort data provide little support for an association between dietary fat and colorectal cancer risk. However, intake of fried foods and/or other factors related to their intake may be associated with colorectal cancer development. This finding warrants further examination. PMID- 15522859 TI - Accuracy and repeatability of commercial geocoding. AB - The authors estimated accuracy and repeatability of commercial geocoding to guide vendor selection in the Life Course Socioeconomic Status, Social Context and Cardiovascular Disease study (2001-2002). They submitted 1,032 participant addresses (97% in Maryland, Minnesota, Mississippi, or North Carolina) to vendor A twice over 9 months and measured repeatability as agreement between levels of address matching, discordance (%) between statistical tabulation areas, and median distance (d, in meters) and bearing (theta;, in degrees) between coordinates assigned on each occasion (H(o):Sigma(i)( = 1 -->) (n) [theta;(i) /n] = 180 degrees ). They also submitted 75 addresses of nearby air pollution monitors (77% urban/suburban; 69% residential/commercial) to vendors A and B and then measured accuracy by comparing vendor- and US Environmental Protection Agency (EPA)-assigned geocodes using the above measures. Repeatability of geocodes assigned by vendor A was high (kappa = 0.90; census block group discordance = 5%; d < 1 m; theta; = 177 degrees ). The match rate for EPA monitor addresses was higher for vendor B versus A (88% vs. 76%), but discordance at census block group, tract, and county levels also was, respectively, 1.4-, 1.9-, and 5.0-fold higher for vendor B. Moreover, coordinates assigned by vendor B were further from those assigned by the EPA (d = 212 m vs. 149 m; theta; = 131 degrees vs. 171 degrees ). These findings suggest that match rates, repeatability, and accuracy should be used to guide vendor selection. PMID- 15522860 TI - Re: "evaluation of mortality data for older Mexican Americans: implications for the Hispanic paradox". PMID- 15522861 TI - Re: "does job strain increase the risk for coronary heart disease or death in men and women? The Framingham offspring study". PMID- 15522863 TI - Crystal structure of a 92-residue C-terminal fragment of TonB from Escherichia coli reveals significant conformational changes compared to structures of smaller TonB fragments. AB - Uptake of siderophores and vitamin B(12) through the outer membrane of Escherichia coli is effected by an active transport system consisting of several outer membrane receptors and a protein complex of the inner membrane. The link between these is TonB, a protein associated with the cytoplasmic membrane, which forms a large periplasmic domain capable of interacting with several outer membrane receptors, e.g. FhuA, FecA, and FepA for siderophores and BtuB for vitamin B(12.) The active transport across the outer membrane is driven by the chemiosmotic gradient of the inner membrane and is mediated by the TonB protein. The receptor-binding domain of TonB appears to be formed by a highly conserved C terminal amino acid sequence of approximately 100 residues. Crystal structures of two C-terminal TonB fragments composed of 85 (TonB-85) and 77 (TonB-77) amino acid residues, respectively, have been previously determined (Chang, C., Mooser, A., Pluckthun, A., and Wlodawer, A. (2001) J. Biol. Chem. 276, 27535-27540 and Koedding, J., Howard, S. P., Kaufmann, L., Polzer, P., Lustig, A., and Welte, W. (2004) J. Biol. Chem. 279, 9978-9986). In both cases the TonB fragments form dimers in solution and crystallize as dimers consisting of monomers tightly engaged with one another by the exchange of a beta-hairpin and a C-terminal beta strand. Here we present the crystal structure of a 92-residue fragment of TonB (TonB-92), which is monomeric in solution. The structure, determined at 1.13-A resolution, shows a dimer with considerably reduced intermolecular interaction compared with the other known TonB structures, in particular lacking the beta hairpin exchange. PMID- 15522862 TI - Kinetic and molecular analysis of 5-epiaristolochene 1,3-dihydroxylase, a cytochrome P450 enzyme catalyzing successive hydroxylations of sesquiterpenes. AB - The final step of capsidiol biosynthesis is catalyzed by 5-epiaristolochene dihydroxylase (EAH), a cytochrome P450 enzyme that catalyzes the regio- and stereospecific insertion of two hydroxyl moieties into the bicyclic sesquiterpene 5-epiaristolochene (EA). Detailed kinetic studies using EA and the two possible monohydroxylated intermediates demonstrated the release of 1beta-hydroxy-EA ((OH)EA) at high EA concentrations and a 10-fold catalytic preference for 1beta(OH)EA versus 3alpha(OH)EA, indicative of a preferred reaction order of hydroxylation at C-1, followed by that at C-3. Sequence alignments and homology modeling identified active-site residues tested for their contribution to substrate specificity and overall enzymatic activity. Mutants EAH-S368C and EAH S368V exhibited wild-type catalytic efficiencies for 1beta(OH)EA biosynthesis, but were devoid of the successive hydroxylation activity for capsidiol biosynthesis. In contrast to EAH-S368C, EAH-S368V catalyzed the relative equal biosynthesis of 1beta(OH)EA, 2beta(OH)EA, and 3beta(OH)EA from EA with wild-type efficiency. Moreover, EAH-S368V converted approximately 1.5% of these monohydroxylated products to their respective ketone forms. Alanine and threonine mutations at position 368 were significantly compromised in their conversion rates of EA to capsidiol and correlated with 3.6- and 5.7-fold increases in their Km values for the 1beta(OH)EA intermediate, respectively. A role for Ile486 in the successive hydroxylations of EA was also suggested by the EAH-I468A mutant, which produced significant amounts 1beta(OH)EA, but negligible amounts of capsidiol from EA. The altered product profile of the EAH-I486A mutant correlated with a 3.6-fold higher Km for EA and a 4.4-fold slower turnover rate (kcat) for 1beta(OH)EA. These kinetic and mutational studies were correlated with substrate docking predictions to suggest how Ser368 and Ile486 might contribute to active site topology, substrate binding, and substrate presentation to the oxo-Fe-heme reaction center. PMID- 15522864 TI - Differential protein mobility of the gamma-aminobutyric acid, type A, receptor alpha and beta subunit channel-lining segments. AB - The gamma-aminobutyric acid, type A (GABAA), receptor ion channel is lined by the second membrane-spanning (M2) segments from each of five homologous subunits that assemble to form the receptor. Gating presumably involves movement of the M2 segments. We assayed protein mobility near the M2 segment extracellular ends by measuring the ability of engineered cysteines to form disulfide bonds and high affinity Zn(2+)-binding sites. Disulfide bonds formed in alpha1beta1E270Cgamma2 but not in alpha1N275Cbeta1gamma2 or alpha1beta1gamma2K285C. Diazepam potentiation and Zn2+ inhibition demonstrated that expressed receptors contained a gamma subunit. Therefore, the disulfide bond in alpha1beta1E270Cgamma2 formed between non-adjacent subunits. In the homologous acetylcholine receptor 4-A resolution structure, the distance between alpha carbon atoms of 20' aligned positions in non-adjacent subunits is approximately 19 A. Because disulfide trapping involves covalent bond formation, it indicates the extent of movement but does not provide an indication of the energetics of protein deformation. Pairs of cysteines can form high affinity Zn(2+)-binding sites whose affinity depends on the energetics of forming a bidentate-binding site. The Zn2+ inhibition IC50 for alpha1beta1E270Cgamma2 was 34 nm. In contrast, it was greater than 100 microM in alpha1N275Cbeta1gamma2 and alpha1beta1gamma2K285C receptors. The high Zn2+ affinity in alpha1beta1E270Cgamma2 implies that this region in the beta subunit has a high protein mobility with a low energy barrier to translational motions that bring the positions into close proximity. The differential mobility of the extracellular ends of the beta and alpha M2 segments may have important implications for GABA-induced conformational changes during channel gating. PMID- 15522865 TI - Functional characterization in vitro of all two-component signal transduction systems from Escherichia coli. AB - Bacteria possess a signal transduction system, referred to as a two-component system, for adaptation to external stimuli. Each two-component system consists of a sensor protein-histidine kinase (HK) and a response regulator (RR), together forming a signal transduction pathway via histidyl-aspartyl phospho-relay. A total of 30 sensor HKs, including as yet uncharacterized putative HKs (BaeS, BasS, CreC, CusS, HydH, RstB, YedV, and YfhK), and a total of 34 RRs, including putative RRs (BaeR, BasR, CreB, CusR, HydG, RstA, YedW, YfhA, YgeK, and YhjB), have been suggested to exist in Escherichia coli. We have purified the carboxyl terminal catalytic domain of 27 sensor HKs and the full-length protein of all 34 RRs to apparent homogeneity. Self-phosphorylation in vitro was detected for 25 HKs. The rate of self-phosphorylation differed among HKs, whereas the level of phosphorylation was generally co-related with the phosphorylation rate. However, the phosphorylation level was low for ArcB, HydH, NarQ, and NtrB even though the reaction rate was fast, whereas the level was high for the slow phosphorylation species BasS, CheA, and CreC. By using the phosphorylated HKs, we examined trans phosphorylation in vitro of RRs for all possible combinations. Trans phosphorylation of presumed cognate RRs by HKs was detected, for the first time, for eight pairs, BaeS-BaeR, BasS-BasR, CreC-CreB, CusS-CusR, HydH-HydG, RstB RstA, YedV-YedW, and YfhK-YfhA. All trans-phosphorylation took place within less than 1/2 min, but the stability of phosphorylated RRs differed, indicating the involvement of de-phosphorylation control. In addition to the trans phosphorylation between the cognate pairs, we detected trans-phosphorylation between about 3% of non-cognate HK-RR pairs, raising the possibility that the cross-talk in signal transduction takes place between two-component systems. PMID- 15522866 TI - Four Na+/H+ exchanger isoforms are distributed to Golgi and post-Golgi compartments and are involved in organelle pH regulation. AB - Four isoforms of the Na+/H+ exchanger (NHE6-NHE9) are distributed to intracellular compartments in human cells. They are localized to Golgi and post Golgi endocytic compartments as follows: mid- to trans-Golgi, NHE8; trans-Golgi network, NHE7; early recycling endosomes, NHE6; and late recycling endosomes, NHE9. No significant localization of these NHEs was observed in lysosomes. The distribution of these NHEs is not discrete in the cells, and there is partial overlap with other isoforms, suggesting that the intracellular localization of the NHEs is established by the balance of transport in and out of the post-Golgi compartments as the dynamic membrane trafficking. The overexpression of NHE isoforms increased the luminal pH of the compartments in which the protein resided from the mildly acidic pH to the cytosolic pH, suggesting that their in vivo function is to regulate the pH and monovalent cation concentration in these organelles. We propose that the specific NHE isoforms contribute to the maintenance of the unique acidic pH values of the Golgi and post-Golgi compartments in the cell. PMID- 15522867 TI - Stimulus-specific induction of a novel nuclear factor-kappaB regulator, IkappaB zeta, via Toll/Interleukin-1 receptor is mediated by mRNA stabilization. AB - We have recently identified an inducible nuclear factor-kappaB (NF-kappaB) regulator, IkappaB-zeta, which is induced by microbial ligands for Toll-like receptors such as lipopolysaccharide and the proinflammatory cytokine interleukin (IL)-1beta but not by tumor necrosis factor (TNF)-alpha. In the present study, we examined mechanisms for stimulus-specific induction of IkappaB-zeta. The analysis of the IkappaB-zeta promoter revealed an essential role for an NF-kappaB binding sequence in transcriptional activation. The activation, however, did not account for the Toll-like receptor/IL-1 receptor-specific induction of IkappaB-zeta, because the promoter analysis and nuclear run-on analysis indicated that its transcription was similarly induced by TNF-alpha. To examine post-transcriptional regulation, we analyzed the decay of IkappaB-zeta mRNA, and we found that it was specifically stabilized by lipopolysaccharide or IL-1beta but not by TNF-alpha. Furthermore, we found that costimulation with TNF-alpha and another proinflammatory cytokine, IL-17, elicited the IkappaB-zeta induction. Stimulation with IL-17 alone did not induce IkappaB-zeta but stabilized its mRNA. Therefore, IkappaB-zeta induction requires both NF-kappaB activation and stimulus-specific stabilization of its mRNA. Because IkappaB-zeta is essential for expression of a subset of NF-kappaB target genes, the stimulus-specific induction of IkappaB-zeta may be of great significance in regulation of inflammatory reactions. PMID- 15522868 TI - Functional and structural analysis of the GABAA receptor alpha 1 subunit during channel gating and alcohol modulation. AB - The substituted cysteine accessibility method has proven useful for investigating structural changes of the gamma-aminobutyric acid type A (GABA(A)) receptor during channel gating and allosteric modulation. In the present study, the surface accessibility and reaction rate of propyl- and hexyl-methanethiosulfonate to cysteine residues introduced into the third transmembrane segment of the GABA(A) receptor alpha(1) subunit were examined. GABA-induced currents in Xenopus oocytes expressing wild type and cysteine mutant GABA(A) receptors were recorded before and after application of methanethiosulfonate (MTS) reagents in the resting, GABA- or alcohol-bound (ethanol or hexanol) states. Our results indicate that a water-filled cavity exists around the Ala(291) and Tyr(294) residues of the third transmembrane segment, in agreement with previous results. Furthermore, our data indicate that a conformational change produced by alcohols (200 mM ethanol or 0.5 mM hexanol) exposure induces the water cavity around the A291C and Y294C residues to extend deeper, causing the A295C and F296C residues to become accessible to the MTS reagents. In addition, exposure of the A291C, Y294C, F296C, and V297C mutants to MTS reagents in the presence of GABA had significant effects on their GABA-induced currents, indicating that the water cavity around A291C and Y294C residues expanded to F296C and V297C by a structural movement caused by GABA binding. Our data show that GABA(A) receptor is a dynamic protein during alcohol modulation and channel gating. PMID- 15522869 TI - Effects of depletion of CREB-binding protein on c-Myc regulation and cell cycle G1-S transition. AB - We recently reported that the transcriptional coactivator and histone acetyltransferase p300 plays an important role in the G(1) phase of the cell cycle by negatively regulating c-myc and thereby preventing premature G(1) exit (Kolli, et al. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 4646-4651; Baluchamy, et al. (2003) Proc. Natl. Acad. Sci. U. S. A. 100, 9524-9529). Because p300 does not substitute for all CREB-binding protein (CBP) functions, we investigated whether CBP also negatively regulates c-myc and prevents premature DNA synthesis. Here, we show that antisense-mediated depletion of CBP in serum-deprived human cells leads to induction of c-myc and that such cells emerge from quiescence without growth factors at a rate comparable with that of p300-depleted cells. The CBP-depleted cells contained significantly reduced levels of the cyclin-dependent kinase inhibitor p21 and low levels of p107 and p130 (but not pRb) phosphorylation, suggesting that these factors, along with elevated levels of c Myc, contribute to induction of DNA synthesis. Antisense c-Myc reversed the phosphorylation of p107 and p130 and the induction of S phase in CBP-depleted cells, indicating that up-regulation of c-myc is directly responsible for the induction of S phase. Furthermore, the serum-stimulated p300/CBP-depleted cells did not traverse beyond S phase, and a significant number of these cells died of apoptosis, which was not related to p53 levels. These cells also contained significantly higher levels of c-Myc compared with normal cells. When c-myc expression was blocked by antisense c-Myc, the apoptosis of the serum-stimulated CBP-depleted cells was reversed, indicating that high levels of c-Myc contribute to apoptosis. Thus, despite their high degree of structural and functional similarities, normal levels of both p300 and CBP are essential for keeping c-myc in a repressed state in G(1) and thereby preventing inappropriate entry of cells into S phase. In addition, both these proteins also provide important functions in coordinated cell cycle progression. PMID- 15522870 TI - Different immunoreactivity against monoclonal antibodies between wild-type and mutant copper/zinc superoxide dismutase linked to amyotrophic lateral sclerosis. AB - Although more than 100 mutations have been identified in the copper/zinc superoxide dismutase (Cu/Zn-SOD) in familial amyotrophic lateral sclerosis (FALS), the mechanism responsible for FALS remains unclear. The finding of the present study shows that FALS-causing mutant Cu/Zn-SOD proteins (FALS mutant SODs), but not wild-type SOD, are barely detected by three monoclonal antibodies (mAbs) in Western blot analyses. The enzyme-linked immunosorbent assay for denatured FALS mutant SODs by dithiothreitol, SDS, or heat treatment also showed a lowered immunoreactivity against the mAbs compared with wild-type SOD. Because all the epitopes of these mAbs are mapped within the Greek key loop (residues 102 115 in human Cu/Zn-SOD), these data suggest that different conformational changes occur in the loop between wild-type and FALS mutant SODs during the unfolding process. Circular dichroism measurements revealed that the FALS mutant SODs are sensitive to denaturation by dithiothreitol, SDS, or heat treatment, but these results do not completely explain the different recognition by the mAbs between wild-type and FALS mutant SODs under the denatured conditions. The study on the conformational changes in local areas monitoring with mAbs may provide a new insight into the etiology of FALS. PMID- 15522871 TI - In vivo and in vitro characterization of novel neuronal plasticity factors identified following spinal cord injury. AB - Following spinal cord injury, there are numerous changes in gene expression that appear to contribute to either neurodegeneration or reparative processes. We utilized high density oligonucleotide microarrays to examine temporal gene profile changes after spinal cord injury in rats with the goal of identifying novel factors involved in neural plasticity. By comparing mRNA changes that were coordinately regulated over time with genes previously implicated in nerve regeneration or plasticity, we found a gene cluster whose members are involved in cell adhesion processes, synaptic plasticity, and/or cytoskeleton remodeling. This group, which included the small GTPase Rab13 and actin-binding protein Coronin 1b, showed significantly increased mRNA expression from 7-28 days after trauma. Overexpression in vitro using PC-12, neuroblastoma, and DRG neurons demonstrated that these genes enhance neurite outgrowth. Moreover, RNAi gene silencing for Coronin 1b or Rab13 in NGF-treated PC-12 cells markedly reduced neurite outgrowth. Coronin 1b and Rab13 proteins were expressed in cultured DRG neurons at the cortical cytoskeleton, and at growth cones along with the pro plasticity/regeneration protein GAP-43. Finally, Coronin 1b and Rab13 were induced in the injured spinal cord, where they were also co-expressed with GAP-43 in neurons and axons. Modulation of these proteins may provide novel targets for facilitating restorative processes after spinal cord injury. PMID- 15522872 TI - Viability of Escherichia coli topA mutants lacking DNA topoisomerase I. AB - The viability of the topA mutants lacking DNA topoisomerase I was thought to depend on the presence of compensatory mutations in Escherichia coli but not Salmonella typhimurium or Shigella flexneri. This apparent discrepancy in topA requirements in different bacteria prompted us to reexamine the topA requirements in E. coli. We find that E. coli strains bearing topA mutations, introduced into the strains by DNA-mediated gene replacement, are viable at 37 or 42 degrees C without any compensatory mutations. These topA(-) cells exhibit cold sensitivity in their growth, however, and this cold sensitivity phenotype appears to be caused by excessive negative supercoiling of intracellular DNA. In agreement with previous results (Zhu, Q., Pongpech, P., and DiGate, R. J. (2001) Proc. Natl. Acad. Sci. U. S. A. 98, 9766-9771), E. coli cells lacking both type IA DNA topoisomerases I and III are found to be nonviable, indicating that the two type IA enzymes share a critical cellular function. PMID- 15522873 TI - Phosphorylation and sulfation of oligosaccharide substrates critically influence the activity of human beta1,4-galactosyltransferase 7 (GalT-I) and beta1,3 glucuronosyltransferase I (GlcAT-I) involved in the biosynthesis of the glycosaminoglycan-protein linkage region of proteoglycans. AB - We determined whether the two major structural modifications, i.e. phosphorylation and sulfation of the glycosaminoglycan-protein linkage region (GlcAbeta1-3Galbeta1-3Galbeta1-4Xylbeta1), govern the specificity of the glycosyltransferases responsible for the biosynthesis of the tetrasaccharide primer. We analyzed the influence of C-2 phosphorylation of Xyl residue on human beta1,4-galactosyltransferase 7 (GalT-I), which catalyzes the transfer of Gal onto Xyl, and we evaluated the consequences of C-4/C-6 sulfation of Galbeta1-3Gal (Gal2-Gal1) on the activity and specificity of beta1,3-glucuronosyltransferase I (GlcAT-I) responsible for the completion of the glycosaminoglycan primer sequence. For this purpose, a series of phosphorylated xylosides and sulfated C-4 and C-6 analogs of Galbeta1-3Gal was synthesized and tested as potential substrates for the recombinant enzymes. Our results revealed that the phosphorylation of Xyl on the C-2 position prevents GalT-I activity, suggesting that this modification may occur once Gal is attached to the Xyl residue of the nascent oligosaccharide linkage. On the other hand, we showed that sulfation on C 6 position of Gal1 of the Galbeta1-3Gal analog markedly enhanced GlcAT-I catalytic efficiency and we demonstrated the importance of Trp243 and Lys317 residues of Gal1 binding site for enzyme activity. In contrast, we found that GlcAT-I was unable to use digalactosides as acceptor substrates when Gal1 was sulfated on C-4 position or when Gal2 was sulfated on both C-4 and C-6 positions. Altogether, we demonstrated that oligosaccharide modifications of the linkage region control the specificity of the glycosyltransferases, a process that may regulate maturation and processing of glycosaminoglycan chains. PMID- 15522874 TI - Structure of the dimeric PufX-containing core complex of Rhodobacter blasticus by in situ atomic force microscopy. AB - We have studied photosynthetic membranes of wild type Rhodobacter blasticus, a closely related strain to the well studied Rhodobacter sphaeroides, using atomic force microscopy. High-resolution atomic force microscopy topographs of both cytoplasmic and periplasmic surfaces of LH2 and RC-LH1-PufX (RC, reaction center) complexes were acquired in situ. The LH2 is a nonameric ring inserted into the membrane with the 9-fold axis perpendicular to the plane. The core complex is an S-shaped dimer composed of two RCs, each encircled by 13 LH1 alpha/beta heterodimers, and two PufXs. The LH1 assembly is an open ellipse with a topography-free gap of approximately 25 A. The two PufXs, one of each core, are located at the dimer center. Based on our data, we propose a model of the core complex, which provides explanation for the PufX-induced dimerization of the Rhodobacter core complex. The QB site is located facing a approximately 25-A wide gap within LH1, explaining the PufX-favored quinone passage in and out of the core complex. PMID- 15522875 TI - Monoallelic expression and asynchronous replication of p120 catenin in mouse and human cells. AB - The number of autosomal mammalian genes subject to random monoallelic expression has been limited to genes highly specific to the function of chemosensory neurons or lymphocytes, making this phenomenon difficult to address systematically. Here we demonstrate that asynchronous DNA replication can be used as a marker for the identification of novel genes with monoallelic expression and identify p120 catenin, a gene involved in cell adhesion, as belonging to this class. p120 is widely expressed; its presence in available cell lines allowed us to address quantitative aspects of monoallelic expression. We show that the epigenetic choice of active allele is clonally stable and that biallelic clones express p120 at twice the level of monoallelic clones. Unlike previous reports about genes of this type, we found that expression of p120 can be monoallelic in one cell type and strictly biallelic in another. We show that in human lymphoblasts, the silencing of one allele is incomplete. These unexpected properties are likely to be wide-spread, as we show that the Tlr4 gene shares them. Identification of monoallelic expression of a nearly ubiquitous gene indicates that this type of gene regulation is more common than previously thought. This has important implications for carcinogenesis and definition of cell identity. PMID- 15522876 TI - Combinatorial transcription of herpes simplex virus and varicella zoster virus immediate early genes is strictly determined by the cellular coactivator HCF-1. AB - The mammalian transcriptional coactivator host cell factor-1 (HCF-1) functions in concert with Oct-1 and VP16 to assemble the herpes simplex virus (HSV) immediate early (IE) transcription enhancer core complexes that mediate the high level transcription of these genes upon infection. Although this transcriptional model has been well characterized in vitro, the requirements and significance of the components have not been addressed. Oct-1 was previously determined to be critical but not essential for HSV IE gene expression. In contrast, RNA interference-mediated depletion of HCF-1 resulted in abrogation of HSV IE gene expression. The HSV IE gene enhancer domain is a model of combinatorial transcription and consists of the core enhancer and multiple binding sites for factors such as Sp1 and GA-binding protein. It was striking that HCF-1 was strictly required for VP16-mediated transcriptional induction via the core enhancer as well as for basal level transcription mediated by GA-binding protein and Sp1. HCF-1 was also found to be essential for the induction of varicella zoster virus IE gene expression by ORF10, the VZV ortholog of the HSV IE transactivator VP16, and the autostimulatory IE62 protein. The critical dependence upon HCF-1 demonstrates that this cellular component is a key factor for control of HSV and VZV IE gene expression by functioning as the common element for distinct factors cooperating at the IE gene enhancers. The requirements for this protein supports the model whereby the regulated transport of HCF-1 from the cytoplasm to the nucleus in sensory neurons may control IE gene expression and reactivation of these viruses from the latent state. PMID- 15522877 TI - FRAT-2 preferentially increases glycogen synthase kinase 3 beta-mediated phosphorylation of primed sites, which results in enhanced tau phosphorylation. AB - Tau is a microtubule-associated protein found primarily in neurons, and its function is regulated by site-specific phosphorylation. Although it is well established that tau is phosphorylated at both primed and unprimed epitopes by glycogen synthase kinase 3 beta (GSK3 beta), how specific proteins that interact with GSK3 beta regulate tau phosphorylation has not been thoroughly examined. Members of the FRAT (frequently rearranged in advanced T-cell lymphoma) protein family have been shown to interact with GSK3 beta, and FRAT-1 has been shown to modulate the activity of GSK3 beta toward tau and other substrates. However, the effects of FRAT-2 on GSK3 beta activity and tau phosphorylation have not been examined. Therefore in this study the effects of FRAT-2 on GSK3 beta activity and tau phosphorylation were examined. In situ, FRAT-2 significantly increased GSK3 beta-mediated phosphorylation of tau at a primed epitope while not significantly affecting the phosphorylation of unprimed sites. Co-immunoprecipitation studies revealed that association of FRAT-2 with GSK3 beta resulted in a significant increase in phosphorylation of a primed substrate but did not alter phosphorylation of an unprimed substrate. Further, in vitro assays using recombinant proteins directly demonstrated that FRAT-2 enhances GSK3 beta mediated phosphorylation of a primed substrate to a greater extent than an unprimed substrate. In addition, FRAT-2 is phosphorylated by GSK3 beta. This is the first demonstration of a protein differentially regulating the activity of GSK3 beta toward primed and unprimed epitopes. PMID- 15522878 TI - Role of tyrosine 441 of interferon-gamma receptor subunit 1 in SOCS-1-mediated attenuation of STAT1 activation. AB - Suppressor of cytokine signaling (SOCS)-1, the key negative regulator of interferon (IFN)-gamma-dependent signaling, is induced in response to IFNgamma. SOCS-1 binds to and inhibits the IFNgamma receptor-associated kinase Janus activated kinase (JAK) 2 and inhibits its function in vitro, but the mechanism by which SOCS-1 inhibits IFNgamma-dependent signaling in vivo is not clear. Upon stimulation, mouse IFNgamma receptor subunit 1 (IFNGR1) is phosphorylated on several cytoplasmic tyrosine residues, and Tyr(419) is required for signal transducer and activator of transcription (STAT) 1 activation in mouse embryo fibroblasts. However, the functions of the other three cytoplasmic tyrosine residues are not known. Here we show that Tyr(441) is required to attenuate STAT1 activation in response to IFNgamma. Several tyrosine to phenylalanine mutants of IFNGR1, expressed at normal levels in stable pools of IFNGR1-null cells, were analyzed for the phosphorylation of STAT1 during a 48-h period, and antiviral activity in response to IFNgamma was also measured. Stronger activation of STAT1 was observed in cells expressing all IFNGR1 variants mutated at Tyr(441), and, consistently, stronger antiviral activity was also observed in these cells. Furthermore, constitutive overexpression of SOCS-1 inhibited IFNgamma-dependent signaling only in cells expressing IFNGR1 variants that included the Tyr(441) mutation. Mutation of Tyr(441) also blocked the ability of SOCS-1 to bind to IFNGR1 and JAK2 in response to IFNgamma and the normal down-regulation of STAT1 activation and antiviral activity. These results, together with data from the literature, suggest a model in which, in response to IFNgamma, phosphorylation of Tyr(441) creates a docking site for SOCS-1, which then binds to JAK2 within the receptor-JAK complex to partially inhibit JAK2 phosphorylation. Furthermore, the virtually complete blockade of STAT1 phosphorylation by overexpressed SOCS-1 in this experiment suggests that the binding of SOCS-1 to Tyr(441) also blocks the access of STAT1 to Tyr(419) and that this effect may be the principal mechanism of inhibition of downstream signaling. PMID- 15522879 TI - Extracellular ATP-induced proliferation of adventitial fibroblasts requires phosphoinositide 3-kinase, Akt, mammalian target of rapamycin, and p70 S6 kinase signaling pathways. AB - Extracellular nucleotides are increasingly recognized as important regulators of growth in a variety of cell types. Recent studies have demonstrated that extracellular ATP is a potent inducer of fibroblast growth acting, at least in part, through an ERK1/2-dependent signaling pathway. However, the contributions of additional signaling pathways to extracellular ATP-mediated cell proliferation have not been defined. By using both pharmacologic and genetic approaches, we found that in addition to ERK1/2, phosphatidylinositol 3-kinase (PI3K), Akt, mammalian target of rapamycin (mTOR), and p70 S6K-dependent signaling pathways are required for ATP-induced proliferation of adventitial fibroblasts. We found that extracellular ATP acting in part through G(i) proteins increased PI3K activity in a time-dependent manner and transient phosphorylation of Akt. This PI3K pathway is not involved in ATP-induced activation of ERK1/2, implying activation of independent parallel signaling pathways by ATP. Extracellular ATP induced dramatic increases in mTOR and p70 S6K phosphorylation. This activation of the mTOR/p70 S6 kinase (p70 S6K) pathway in response to ATP is because of independent contributions of PI3K/Akt and ERK1/2 pathways, which converge on the level of p70 S6K. ATP-dependent activation of mTOR and p70 S6K also requires additional signaling inputs perhaps from pathways operating through Galpha or Gbetagamma subunits. Collectively, our data demonstrate that ATP-induced adventitial fibroblast proliferation requires activation and interaction of multiple signaling pathways such as PI3K, Akt, mTOR, p70 S6K, and ERK1/2 and provide evidence for purinergic regulation of the protein translational pathways related to cell proliferation. PMID- 15522880 TI - Interleukin-12-induced interferon-gamma production by human peripheral blood T cells is regulated by mammalian target of rapamycin (mTOR). AB - Depending on the type of external signals, T cells can initiate multiple intracellular signaling pathways that can be broadly classified into two groups based on their sensitivity to the immunosuppressive drug cyclosporin A (CsA). Interleukin (IL)-12-mediated interferon (IFN)-gamma production by activated T cells has been shown to be CsA-insensitive. In this report, we demonstrate that the IL-12-induced CsA-resistant pathway of IFN-gamma production is sensitive to rapamycin. Rapamycin treatment resulted in the aberrant recruitment of Stat3, Stat4, and phospho-c-Jun to the genomic promoter region resulting in decreased IFN-gamma transcription. IL-12-induced phosphorylation of Stat3 on Ser-727 was affected by rapamycin, which may be due to the effect of rapamycin on the IL-12 induced interaction between mammalian target of rapamycin (mTOR) and Stat3. In accordance with this, reduction in the mTOR protein level by small interfering RNA resulted in suppression of Stat3 phosphorylation and decreased production of IFN-gamma after IL-12 stimulation. These results suggest that mTOR may play a major role in IL-12-induced IFN-gamma production by activated T cells. PMID- 15522881 TI - Characterization of the Alzheimer's disease-associated CLAC protein and identification of an amyloid beta-peptide-binding site. AB - Amyloid beta-peptide (Abeta) deposition into amyloid plaques is one of the invariant neuropathological features of Alzheimer's disease. Other proteins co deposit with Abeta in plaques, and one recently identified amyloid-associated protein is the collagen-like Alzheimer amyloid plaque component CLAC. It is not known how CLAC deposition affects Abeta plaque genesis and the progress of the disease. Here, we studied the in vitro properties of CLAC purified from a mammalian expression system. CLAC displays features characteristic of a collagen protein, e.g. it forms a partly protease-resistant triple-helical structure, exhibits an intermediate affinity for heparin, and is glycosylated. Purified CLAC was also used to investigate the interaction between CLAC and Abeta. Using a solid-phase binding assay, we show that CLAC bound with a similar affinity to aggregates formed by Abeta-(1-40) and Abeta-(1-42) and that the interaction was impaired by increasing salt concentrations. An 8-residue-long sequence located in non-collagenous domain 2 of CLAC was found to be crucial for the interaction with Abeta. These findings may be useful for future therapeutic interventions aimed at finding compounds that modulate the binding of CLAC to Abeta deposits. PMID- 15522882 TI - Apoptosis and survival of osteoblast-like cells are regulated by surface attachment. AB - We tested the hypothesis that RGDS peptides regulate osteoblast survival in culture. Osteoblast-like MC3T3-E1 cells were allowed to attach to RGDS peptides that had been tethered to a silicone surface utilizing a previously described grafting technique. The RGDS-modified surface caused up-regulation of alpha(v)beta(3) integrin. We noted that there was an increase in expression of activated focal adhesion kinase and activated Akt. There was no change in the expression level of the anti-apoptotic protein Bcl-2, the pro-apoptotic protein Bad, or the inactivated form of Bad, pBad. Attachment to the RGDS-treated membrane completely abolished apoptosis induced by staurosporine, the Ca(2+).P(i) ion pair, and sodium nitroprusside. However, the surface modification did not interfere with apoptosis mediated by the free RGDS peptide or serum-free medium. When the activity of the phosphatidylinositol 3-kinase pathway was inhibited, RGDS-dependent resistance to apoptosis was eliminated. These results indicated that the binding of cells to RGDS abrogated apoptosis via the mitochondrial pathway and that the suppression of apoptosis was dependent on the activity of phosphatidylinositol 3-kinase. PMID- 15522883 TI - Identification of the namH gene, encoding the hydroxylase responsible for the N glycolylation of the mycobacterial peptidoglycan. AB - The peptidoglycan of most bacteria consists of a repeating disaccharide unit of beta-1,4-linked N-acetylmuramic acid and N-acetylglucosamine. However, the muramic acid moieties of the mycobacterial peptidoglycan are N-glycolylated, not N-acetylated. This is a rare modification seen only in the peptidoglycan of mycobacteria and five other closely related genera of bacteria. The N glycolylation of sialic acids is a unique carbohydrate modification that has been studied extensively in eukaryotes. However, the significance of the N glycolylation of bacterial peptidoglycan is unknown. The goal of this project was to identify the gene encoding the hydroxylase responsible for the N-glycolylation of the mycobacterial peptidoglycan. We developed a novel assay for the mycobacterial UDP-N-acetylmuramic acid hydroxylation reaction and demonstrated that Mycobacterium smegmatis has an enzyme activity that can convert UDP-N acetylmuramic acid to UDP-N-glycolylmuramic acid. We identified the gene namH encoding the mycobacterial UDP-N-acetylmuramic acid hydroxylase by computer data base searching and motif comparisons with the eukaryotic enzymes responsible for the N-glycolyation of sialic acids. The namH gene is not essential for in vitro growth as we were successful in deleting the gene in M. smegmatis. The M. smegmatis mutant is devoid of UDP-N-acetylmuramic acid hydroxylase activity and synthesizes only N-acetylated muropeptide precursors. Furthermore, the mutant exhibits increased susceptibility to beta-lactam antibiotics and lysozyme. Our studies suggest that the N-glycolylation of mycobacterial peptidoglycan may play a role in lysozyme resistance or may contribute to the structural stability of the cell wall architecture. PMID- 15522884 TI - Domain V of beta2-glycoprotein I binds factor XI/XIa and is cleaved at Lys317 Thr318. AB - The fifth domain (DV) of beta2-glycoprotein I (beta2GPI) is important for binding a number of ligands including phospholipids and factor XI (FXI). Beta2GPI is proteolytically cleaved in DV by plasmin but not by thrombin, VIIa, tissue plasminogen activator, or uPA. Following proteolytic cleavage of DV by plasmin, beta2GPI retains binding to FXI but not to phospholipids. Native beta2GPI, but not cleaved beta2GPI, inhibits activation of FXI by thrombin and factor XIIa, attenuating a positive feedback mechanism for additional thrombin generation. In this report, we have defined the FXI/FXIa binding site on beta2GPI using site directed mutagenesis. We show that the positively charged residues Lys284, Lys286, and Lys287 in DV are essential for the interaction of beta2GPI with FXI/FXIa. We also demonstrate that FXIa proteolytically cleaves beta2GPI at Lys317-Thr318 in DV. Thus, FXIa cleavage of beta2GPI in vivo during thrombus formation may accelerate FXI activation by decreasing the inhibitory effect of beta2GPI. PMID- 15522885 TI - Myosin 3A transgene expression produces abnormal actin filament bundles in transgenic Xenopus laevis rod photoreceptors. AB - Myo3A, a class III myosin, localizes to the distal (plus) ends of inner segment actin filament bundles that form the core of microvillus-like calycal processes encircling the base of the photoreceptor outer segment. To investigate Myo3A localization and function, we expressed green fluorescent protein-tagged bass Myo3A and related constructs in transgenic Xenopus rods using a modified opsin promoter. Tagged intact Myo3A localized to rod calycal processes, as previously reported for native bass Myo3A. Transgenic rods developed abnormally large calycal processes and subsequently degenerated. Modified Myo3A expression constructs demonstrated that calycal process localization required an active motor domain and the tail domain. Expressed tail domain alone localized to actin bundles along the entire inner segment length, rather than to the distal end. This tail domain localization required the conserved C-terminal domain (3THDII) previously shown to possess an actin-binding motif. Our findings suggest that Myo3A plays a role in the morphogenesis and maintenance of calycal processes of vertebrate photoreceptors. PMID- 15522886 TI - Ca2+-dependent and -independent mechanisms of calmodulin nuclear translocation. AB - Translocation from the cytosol to the nucleus is a major response by calmodulin (CaM) to stimulation of cells by Ca2+. However, the mechanisms involved in this process are still controversial and both passive and facilitated diffusion have been put forward. We tested nuclear translocation mechanisms in electroporated HeLa cells, rat cortical neurons and glial cells using novel calmodulin and inhibitor peptide probes and confocal microscopy. Passive diffusion of calmodulin across the nuclear membrane was measured in conditions in which facilitated transport was blocked and was compared to that of a similarly sized fluorescein labeled dextran. Wheat germ agglutinin, which blocks facilitated transport but not passive diffusion, inhibited the nuclear entry of both wild-type and Ca2+ binding-deficient mutant calmodulin both in low and elevated [Ca2+]. Ca2+ dependent nuclear translocation was prevented by a membrane-permeant CaM inhibitor, the mTrp peptide, which indicated that it was specific to Ca2+/CaM. Diffusion of free CaM and Ca2+/CaM was considerably slower than the observed nuclear translocation by facilitated transport. Our data show that the majority of CaM nuclear entry occurred by facilitated mechanisms in all cell types examined, in part by a Ca2+-independent and in part by a Ca2+-dependent translocation mechanism. PMID- 15522887 TI - Functional characterization of a mouse testicular olfactory receptor and its role in chemosensing and in regulation of sperm motility. AB - Although a subset of the olfactory receptor (OR) gene family is expressed in testis, neither their developmental profile nor their physiological functions have been fully characterized. Here, we show that MOR23 (a mouse OR expressed in the olfactory epithelium and testis) functions as a chemosensing receptor in mouse germ cells. In situ hybridization showed that MOR23 was expressed in round spermatids during stages VI-VIII of spermatogenesis. Lyral, a cognate ligand of MOR23, caused an increase in intracellular Ca2+ in a fraction of spermatogenic cells and spermatozoa. We also generated transgenic mice that express high levels of MOR23 in the testis and examined the response of their germ cells to lyral. The results provided evidence that lyral-induced Ca2+ increases were indeed mediated by MOR23. In a sperm accumulation assay, spermatozoa migrated towards an increasing gradient of lyral. Tracking and sperm flagellar analyses suggest that Ca2+ increases caused by MOR23 activation lead to modulation of flagellar configuration, resulting in chemotaxis. By contrast, a gradient of a cAMP analog or K8.6 solution, which elicit Ca2+ influx in spermatozoa, did not cause sperm accumulation, indicating that chemosensing and regulation of sperm motility was due to an OR-mediated local Ca2+ increase. The present studies indicate that mouse testicular ORs might play a role in chemoreception during sperm-egg communication and thereby regulate fertilization. PMID- 15522888 TI - Migration and retraction of endothelial and epithelial cells require PHI-1, a specific protein-phosphatase-1 inhibitor protein. AB - Cell migration and retraction are interrelated activities that are crucial for a range of physiological processes such as wound healing and vascular permeability. Immunostaining of brain sections for the specific inhibitor of type-1 protein Ser/Thr phosphatase called PHI-1 showed high expression levels in smooth muscle and especially in vascular endothelial cells. During migration of cultured human lung microvascular endothelial cells, endogenous PHI-1 was concentrated to the trailing edge of the cells. Knockdown of PHI-1 using small interfering RNAs reduced by 45% the rate of HeLa cell migration in a wound-healing assay. These cells exhibited an extremely elongated phenotype relative to controls and time lapse movies revealed a defect in retraction of the trailing edge. Both HeLa and human vascular endothelial cells depleted of PHI-1 showed increased surface areas relative to controls during cell spreading in a replating assay. Analysis of sequential microscopic images demonstrated this was due to a significant decrease in the number of retraction events, whereas protrusive action was unaffected. The Ser/Thr phosphorylation of several signaling, cytoskeletal and focal-adhesion proteins was unchanged in PHI-1-depleted cells, so the target of PHI-1 inhibited protein-phosphatase 1 remains unidentified. Nonetheless, the results show that PHI-1 participates in regulatory events at the trailing edge of migrating cells and modulates retraction of endothelial and epithelial cells. PMID- 15522889 TI - GSK-3beta inhibition/beta-catenin stabilization in ventral midbrain precursors increases differentiation into dopamine neurons. AB - Wnts are important regulators of dopamine (DA) neuron differentiation in the developing ventral mesencephalon and could thus serve as potential tools in the treatment of Parkinson's disease. In this study, we investigate whether established intracellular Wnt signalling components could modulate the development of DA neurons. Two chemical inhibitors of glycogen synthase kinase (GSK)-3beta, indirubin-3-monoxime and kenpaullone, were found to increase neuronal differentiation in ventral mesencephalon precursor cultures. In addition, the GSK-3beta-specific inhibitor kenpaullone increased the size of the DA neuron population through conversion of precursors expressing the orphan nuclear receptor-related factor 1 into tyrosine hydroxylase positive neurons, thereby mimicking an effect of Wnts. We show that GSK-3beta inhibitors stabilized beta-catenin and that overexpression of beta-catenin in ventral mesencephalic precursors resulted in increased DA differentiation. The three- to fivefold increase in DA differentiation of precursor cells by GSK-3beta inhibitors suggests that such compounds could be used to improve stem/precursor cell therapy approaches in Parkinson's disease. PMID- 15522890 TI - Organization and pairing of meiotic chromosomes in the ciliate Tetrahymena thermophila. AB - During meiotic prophase in the ciliate Tetrahymena thermophila micronuclei dramatically elongate and form thread-like crescents. The arrangement of the chromosomes within the crescent as well as the timing of chromosome pairing and recombination with respect to the elongation process have been subjects of ongoing debate. Here, we addressed these issues by means of fluorescence in situ hybridization, labeling of individual chromosomes by BrdU (BrdU-painting) and by immunostaining of the recombination protein, Rad51. BrdU-painting indicated that chromosomes are arranged as parallel bundles within the crescent, and telomere directed fluorescent in situ hybridization (FISH) revealed that most if not all telomeres are assembled near one end of the developing crescent. Prior to full crescent formation, Rad51 localizes to chromatin as numerous foci. Locus-specific FISH demonstrated that close pairing of homologues only occurs in the full crescent. Meiotic DNA double-strand break formation and the initiation of recombination thus seem to precede close pairing. A synaptonemal complex was not detected. We conclude that the chromosomes adopt a polarized arrangement within the crescent, probably resembling the classical bouquet arrangement. Furthermore, we propose that the elongated shape of meiotic micronuclei promotes the parallel arrangement of chromosomes and supports the juxtaposition of homologous regions in the absence of a synaptonemal complex. Several pieces of evidence indicate the presence of one to four chiasmata per bivalent, which would call for crossover interference to explain regular bivalent formation in spite of this low mean number. Tetrahymena might, therefore, pose a case of interference in the absence of a synaptonemal complex. PMID- 15522891 TI - DNA replication licensing in somatic and germ cells. AB - The DNA replication (or origin) licensing system ensures precise duplication of the genome in each cell cycle and is a powerful regulator of cell proliferation in metazoa. Studies in yeast, Drosophila melanogaster and Xenopus laevis have characterised the molecular machinery that constitutes the licensing system, but it remains to be determined how this important evolutionary conserved pathway is regulated in Homo sapiens. We have investigated regulation of the origin licensing factors Cdc6, Cdt1, Mcm2 and Geminin in human somatic and germ cells. Cdc6 and Cdt1 play an essential role in DNA replication initiation by loading the Mcm2-7 complex, which is required for unwinding the DNA helix, onto chromosomal origins. Geminin is a repressor of origin licensing that blocks Mcm2-7 loading onto origins. Our studies demonstrate that Cdc6, Cdt1 and Mcm2 play a central role in coordinating growth during the proliferation-differentiation switch in somatic self-renewing systems and that Cdc6 expression is rate-limiting for acquisition of replication competence in primary oocytes. In striking contrast, we show that proliferation control during male gametogenesis is not linked to Cdc6 or Mcm2, but appears to be coordinated by the negative regulator Geminin with Cdt1 becoming rate-limiting in late prophase. Our data demonstrate a striking sexual dimorphism in the mechanisms repressing origin licensing and preventing untimely DNA synthesis during meiosis I, implicating a pivotal role for Geminin in maintaining integrity of the male germline genome. PMID- 15522892 TI - Mammalian GRIP domain proteins differ in their membrane binding properties and are recruited to distinct domains of the TGN. AB - The four mammalian golgins, p230/golgin-245, golgin-97, GCC88 and GCC185 are targeted to trans-Golgi network (TGN) membranes by their C-terminal GRIP domain in a G-protein-dependent process. The Arf-like GTPase, Arl1, has been shown to mediate TGN recruitment of p230/golgin245 and golgin-97 by interaction with their GRIP domains; however, it is not known whether all the TGN golgins bind to Arl1 and whether they are all recruited to the same or different TGN domains. Here we demonstrate differences in membrane binding properties and TGN domain recruitment of the mammalian GRIP domain proteins. Overexpression of full-length GCC185 resulted in the appearance of small punctate structures dispersed in the cytoplasm of transfected cells that were identified as membrane tubular structures by immunoelectron microscopy. The cytoplasmic GCC185-labelled structures were enriched for membrane binding determinants of GCC185 GRIP, whereas the three other mammalian GRIP family members did not colocalize with the GCC185-labelled structures. These GCC185-labelled structures included the TGN resident protein alpha2,6 sialyltransferase and excluded the recycling TGN protein, TGN46. The Golgi stack was unaffected by overexpression of GCC185. Overexpression of both full-length GCC185 and GCC88 showed distinct and nonoverlapping structures. We also show that the GRIP domains of GCC185 and GCC88 differ in membrane binding properties from each other and, in contrast to p230/golgin-245 and golgin-97, do not interact with Arl1 in vivo. Collectively these results show that GCC88, GCC185 and p230/golgin245 are recruited to functionally distinct domains of the TGN and are likely to be important for the maintenance of TGN subdomain structure, a critical feature for mediating protein sorting and membrane transport. PMID- 15522893 TI - Adhesion-contractile balance in myocyte differentiation. AB - Tissue cells generally pull on their matrix attachments and balance a quasi static contractility against adequate adhesion, but any correlation with and/or influence on phenotype are not yet understood. Here, we begin to demonstrate how differentiation state couples to actomyosin-based contractility through adhesion and substrate compliance. Myotubes are differentiated from myoblasts on collagen patterned coverslips that allow linear fusion but prevent classic myotube branching. Post-fusion, myotubes adhere to the micro-strips but lock into a stress fiber-rich state and do not differentiate significantly further. In contrast, myotubes grown on top of such cells do progress through differentiation, exhibiting actomyosin striations within one week. A compliant adhesion to these lower cells is suggested to couple to contractility and accommodate the reorganization needed for upper cell striation. Contractility is assessed in these adherent cells by mechanically detaching one end of the myotubes. All myotubes, whether striated or not, shorten with an exponential decay. The cell-on-cell myotubes relax more, which implies a greater contractile stress. The non-muscle myosin II inhibitor blebbistatin inhibits relaxation for either case. Myotubes in culture are thus clearly prestressed by myosin II, and this contractility couples to substrate compliance and ultimately influences actomyosin striation. PMID- 15522894 TI - PG-M/versican binds to P-selectin glycoprotein ligand-1 and mediates leukocyte aggregation. AB - P-selectin glycoprotein ligand-1 (PSGL-1), a glycoprotein expressed on the cell surface of leukocytes, binds to selectins and mediates leukocyte rolling on the vascular endothelium. Here we report that PSGL-1 binds to the C-terminal (G3 domain) of the extracellular proteoglycan PG-M/versican. Cells transfected with PSGL-1 or a shorter form containing the binding site, or cells expressing endogenous PSGL-1 aggregate in the presence of versican or G3 product. The aggregation appears to be induced by G3 multimers that bind to PSGL-1 and form a network. Endogenous versican and/or G3-containing fragments also bind to PSGL-1 in human plasma. Removal of the endogenous G3-containing fragments reduces the effect of plasma on leukocyte aggregation. Finally, the roles of G3-containing fragments in leukocyte aggregation were confirmed in a mouse model. Taken together, our results strongly support a physiologically relevant role for PSGL 1/versican binding and may have implications in the immunoresponse. PMID- 15522895 TI - A variant histone H3 is enriched at telomeres in Trypanosoma brucei. AB - Variant histones play critical roles in transcriptional activation and repression, DNA repair and chromosome segregation. We have identified HTV, a single-copy gene in Trypanosoma brucei encoding a variant form of histone H3 (H3V). H3V is present at discrete nuclear foci that shift over the course of the cell cycle and associate with the mitotic spindle, a pattern of localization reminiscent of that described previously for both mini-chromosomes and telomeres. By combining fluorescence in situ hybridization with indirect immunofluorescence, we confirmed that the H3V foci overlap with a 177-bp repetitive sequence element found predominantly in mini-chromosomes, as well as with the TTAGGG repeats that compose telomeres. Chromatin immunoprecipitation studies, however, reveal that only the telomeric repeat DNA is substantially enriched with H3V. HTV is not essential for viability, mini-chromosome segregation, telomere maintenance or transcriptional silencing at the telomere-proximal expression sites from which bloodstream-form T. brucei controls antigenic variation. We propose that H3V represents a novel class of histone H3 variant, a finding that has evolutionary implications. PMID- 15522896 TI - Accumulation of type IV collagen in dilated ER leads to apoptosis in Hsp47 knockout mouse embryos via induction of CHOP. AB - Hsp47 is an endoplasmic reticulum (ER)-resident molecular chaperone that is specific for collagen. In Hsp47(-/-) mouse embryos at 9.5 days postcoitus (dpc), immunostaining indicated the absence of type IV collagen, but not of laminin and nidogen-1, in the basement membrane (BM). Electron immunomicroscopy revealed accumulation of type IV collagen in dilated ERs, but not in the BM of Hsp47(-/-) embryos, whereas it was only present in the BM in Hsp47(+/+) embryos. The BM structures stained with anti-laminin and anti-nidogen-1 antibody became disrupted in Hsp47(-/-) embryos at 10.5 dpc. Thus, in the absence of type IV collagen in the BM owing to the lack of Hsp47, the structure of the BM cannot be maintained during the dramatic morphological changes that take place around 10.5 dpc. Type IV collagen is therefore indispensable for the maintenance of BM structures during the late-stage development of mouse embryos, although not essential for the initial formation of the BM. Just before the death of Hsp47(-/-) embryos, DNA fragmentation typical of apoptosis was observed at 10.5 dpc together with significantly upregulated CHOP mRNA expression. ER stress caused by the accumulation of misfolded collagen may have induced apoptosis in Hsp47-knockout embryos through the upregulation of CHOP. PMID- 15522897 TI - Design and testing of an interactive smoking cessation intervention for inner city women. AB - The purpose of this study was to design and test the usability of a computer mediated smoking cessation program for inner-city women. Design and content were developed consistent with principles of user-centered design. Formative and summative evaluation strategies were utilized in its testing. The summative evaluation was designed to test usability in a naturalistic environment. A sample of 100 women who receive care at an inner-city community health center participated in the study. Average time for completing the computer program was 13.9 minutes. Participants reported a high level of satisfaction with usability of the program. Standardized instruments to measure cognitive processes of change related to smoking were completed at baseline and at 1 week. Participants reported a decrease in favorable attitudes toward smoking (P=0.014) and an increase in cognitive change processes at follow-up (P=0.037). These results indicate that interactive computer technology is acceptable to, and potentially useful for, promoting smoking cessation in low-income women. PMID- 15522898 TI - Quality of implementation: developing measures crucial to understanding the diffusion of preventive interventions. AB - As prevention programs become disseminated, the most serious threat to effectiveness is maintaining the quality of implementation intended by the developers. This paper proposes a methodology for measuring quality of implementation in school settings and presents data from a pilot study designed to test several of the proposed components. These methods included assessments of adherence, quality of process, the positive or negative valence of adaptations, teachers' attitudes and teachers' understanding of program content. This study was conducted with 11 teachers who had varying degrees of experience who taught Life Skills Training. Observation and interview data were collected during visits to schools. Results suggest that quality of implementation can be measured through observation and interview. Teachers varied in adherence and quality of program delivery. All teachers made adaptations to the program. Experienced teachers were more likely to adhere to the curriculum, deliver it in a way that was more interactive and engaging to students, communicate the goals and objectives better, and make positive adaptations. The field can use these findings as the basis for exploring strategies for measuring and improving quality of implementation. PMID- 15522899 TI - Sodium ferric gluconate causes oxidative stress but not acute renal injury in patients with chronic kidney disease: a pilot study. AB - BACKGROUND: Intravenous (i.v) iron is widely used to treat anaemia in patients with chronic kidney disease (CKD). Although beneficial and usually well tolerated, concerns have been raised about its ability to cause oxidative stress and renal injury. METHODS: To determine if i.v. iron causes oxidative stress [as assessed by plasma and urine malondialdehye (MDA)] and/or renal injury (as assessed by urinary albumin, total protein and enzymuria), we conducted a prospective, four-way randomized crossover, blinded end-point trial in eight patients with CKD. Two widely used doses of sodium ferric gluconate (125 mg infused over 1 h and 250 mg infused over 2 h) were given with or without the antioxidant N-acetylcysteine (NAC), resulting in four treatment dose antioxidant/placebo combinations in each patient. Transferrin saturation was measured with urea polyacrylamide gel electrophoresis, MDA by high performance liquid chromatography, and albuminuria and proteinuria by standard clinical methods. Enzymuria was assessed by measurement of N-acetyl-beta-D-glucosaminidase (NAG) excretion by colorimetric assay. RESULTS: I.v. ferric gluconate infusion at both doses resulted in a marked increase in transferrin saturation and a significant increase in plasma MDA levels. Urinary MDA levels also increased at the higher dose of iron. There was no evidence of acute renal injury, as assessed by albuminuria, proteinuria or enzymuria. Pre-treatment with NAC had no effect on oxidative stress or the above urinary parameters. CONCLUSIONS: I.v. ferric gluconate caused oxidative stress (as reflected by increased MDA), but this was not associated with biochemical manifestations of acute renal injury. PMID- 15522900 TI - High molecular weight plasma proteins induce apoptosis and Fas/FasL expression in human proximal tubular cells. AB - BACKGROUND: In proteinuria, proximal tubular epithelial cells (PTECs) are exposed to abnormally high protein concentrations, eventually leading to tubular atrophy and end-stage renal disease. The mode of cell death leading to tubular atrophy in proteinuria has not been fully established. This study examines the role of protein overload on apoptosis, necrosis and cell proliferation in primary cultures of human PTECs using plasma protein fractions representative of selective and non-selective proteinuria. The involvement of the Fas/Fas ligand (FasL) system was also investigated. METHODS: Plasma was collected from healthy volunteers and fractionated into albumin-rich (30-100 kDa), high molecular weight (100-440 kDa) and combined (30-440 kDa) fractions. PTECs were exposed to 10 mg/ml of the protein fractions for 24, 48 and 72 h. Apoptosis was measured using fluorescein isothiocyanate (FITC)-annexinV and TUNEL. Necrosis was measured using propidium iodide, metabolic activity by MTT and cell proliferation by bromodeoxyuridine incorporation. Fas and FasL expression was analysed by western blotting. RESULTS: Exposure to the 100-440 and 30-440 kDa fractions produced significant increases in apoptosis at all time points, whereas PTECs exposed to the 30-100 kDa fraction were not significantly different from control cells. There were no changes in the rates of necrosis as a result of protein loading. A significant reduction in metabolic activity was observed in PTECs exposed to the 100-440 and 30-440 kDa fractions, but not to the 30-100 kDa fraction. Cell proliferation was significantly reduced by 24 h in cells exposed to the 100-440 and 30-440 kDa fractions. By 48 and 72 h, all the three fractions had inhibited cell proliferation. PTECs exposed to the 100-440 and the 30-440 kDa fractions showed a significant upregulation in the expression of Fas and FasL. Overall, the high molecular weight fraction was more 'toxic' than the albumin-rich or combined fraction. CONCLUSION: Increased apoptosis and decreased cell proliferation are the major mechanisms of cell death in human PTECs in response to protein overload. These effects may be mediated at least in part by overexpression of the Fas/FasL system. The severity of such changes is largely determined by the high molecular weight fraction (100-440 kDa) rather than the albumin-rich fraction. PMID- 15522901 TI - The effects of a low-to-moderate intensity pre-conditioning exercise programme linked with exercise counselling for sedentary haemodialysis patients in The Netherlands: results of a randomized clinical trial. AB - OBJECTIVES: The purpose of this study is to determine whether a low-to-moderate intensity pre-conditioning exercise programme linked with exercise counselling could improve behavioural change, physical fitness, physiological condition and health-related quality of life of sedentary haemodialysis patients in The Netherlands. METHODS: Ninety-six haemodialysis patients of the Groningen Dialysis Center were randomized into an exercise group (n = 53) and a control group (n = 43). The exercise programme consists of cycling during dialysis together with a pre-dialysis strength training programme lasting 12 weeks. The intensity of the exercise programme is condition level 12-16 according to the rate of perceived exertion (RPE). Motivational interviewing techniques were used for exercise counselling. Before and after the intervention, both groups were tested on behavioural change and physical fitness components such as reaction time, manual dexterity, lower extremity muscle strength and VO2 peak. Physiological conditions such as weight, blood pressure, haemoglobin and haematocrit values, cholesterol and Kt/V were obtained from the medical records. Health-related quality of life assessment included RAND-36 scores, symptoms and depression. RESULTS: A group x time analysis with MANOVA (repeated measures) demonstrates that participation in a low-to-moderate intensity exercise programme linked with exercise counselling yields a significant increase in behavioural change, reaction time, lower extremity muscle strength, Kt/V and three components of quality of life, and no significant effects in the control group. CONCLUSION: Participating in a low-to moderate intensity pre-conditioning exercise programme showed beneficial effects on behavioural change, physical fitness, physiological conditions and health related quality of life. PMID- 15522902 TI - Embolization of haemodialysis arteriovenous fistulas complicated by venous hypertension: a feasibility study. PMID- 15522903 TI - Mid-dilution on-line haemodiafiltration in a standard dialyser configuration. AB - BACKGROUND: Mid-dilution haemodiafiltration (HDF) results in an improved middle molecule removal compared with standard HDF. The OLpur MD 190 haemodiafilter represents a new dialyser design exclusively for mid-dilution on-line HDF. Compared with standard haemodialysers, structural changes in the headers allow the infusion of high replacement fluid volumes after a first post-dilution and before a second pre-dilution stage. METHODS: We compared in vitro the new device [blood flow (QB) 400 ml/min, substitution flow (QS) 100 and 200 ml/min, dialysate flow (QD) 800 ml/min] with a conventional high-flux dialyser of the same surface area in haemodialysis (HD) (QD 500 ml/min) and post-dilution HDF (at QS 60, QD = 500 ml/min and at QS 100, QD = 800 ml/min) modes. Subsequently, we performed an initial clinical application of the new device in six mid-dilution HDF treatments of five end-stage renal disease patients (QB 400 ml/min, QS 200 ml/min, QD 800 ml/min, treatment duration 205+/-23 min). RESULTS: In vitro urea and beta2 microglobulin clearances in mid-dilution HDF were, respectively, 309.2+/-5.5 and 144.4+/-15.2 ml/min (QS 100) and 321.6+/-4.1 and 204.9+/-4.1 ml/min (QS 200), compared with 278.6+/- 17.2 and 94.0+/-7.6 ml/min in HD, and 310.8+/-10.2 and 123.0+/-6.5 ml/min (QS 60) and 323.6+/-11.2 and 158.0+/-10.3 ml/min (QS 100) in post-dilution HDF. The in vivo trials showed the clinical utility of the device and confirmed the in vitro data: urea and beta2-microglobulin clearances were, respectively, 324.6+/- 10.9 and 207.9+/-29.3 ml/min, while reduction ratios were 75.0+/-5.5 and 83.6+/-4.7%. CONCLUSION: Our preliminary results need confirmation in a prospective cross-over study. However, the Nephros MD 190 haemodiafilter promises to be a true technological step ahead in terms of improved beta2 microglobulin removal. PMID- 15522904 TI - Expanding the donor pool to increase renal transplantation. AB - INTRODUCTION: The goal of the Eurotransplant renal allocation scheme is to provide every patient on the waiting list with a reasonably balanced opportunity for a donor offer. New initiatives were taken in order to maximize donor usage while maintaining a successful transplant outcome. METHODS: Two Eurotransplant projects were launched in order to accommodate changes in donor and recipient profiles. A re-addressing of the non-heart-beating donor pool was undertaken and an allocation scheme in which organs from donors aged >65 are allocated to recipients aged >65 [the Eurotransplant Senior Programme (ESP)] was introduced. RESULTS: Especially in The Netherlands, an enormous increase in the number of non heart-beating donor kidneys has been observed, however with a pace-keeping reduction in heart-beating donors. The organization-wide implementation of the ESP has been successful. The 3 year graft survival rates for these age-matched transplants were as good as the human leukocyte antigen (HLA)-matched transplants (64 vs 67%) (P = 0.4). CONCLUSION: Within the framework of sound research, the utmost flexibility and creativity is needed to keep or even increase the number of renal transplants when faced with a quantitatively stagnating but qualitatively deteriorating donor pool. Both the non-heart-beating donor protocol and the ESP have proven to be quite successful in achieving this goal without compromising the outcome for the individual end-stage renal disease patient. PMID- 15522905 TI - A high-fat diet leads to the progression of non-alcoholic fatty liver disease in obese rats. AB - Fatty livers of obese fa/fa rats are vulnerable to injury when challenged by insults such as endotoxin, ischemia-reperfusion or acute ethanol treatment. The objective of this study was to evaluate whether a high-fat diet can act as a "second hit" and cause progression to liver injury in obese fa/fa rats compared with lean Fa/? rats. Accordingly, obese fa/fa rats and their lean littermates were fed a diet low in fat (12% of total calories) or a diet with 60% calories as lard for 8 weeks. Hyperglycemia and steatohepatitis occurred in the fa/fa rats fed the high-fat diet. This was accompanied by liver injury as assessed by alanine aminotransferase, hematoxilin and eosin staining, increased TNFalpha and stellate cell-derived TGFbeta, collagen deposition, and up-regulation of alpha smooth muscle actin. Active MMP13 decreased in fa/fa rats independently of the diet, and TIMP1 expression increased with the high-fat diet, especially in fa/fa rats. Although UCP2 expression was higher in fa/fa rats regardless of the diet, minor changes in ATP levels were observed. Oxidative stress occurred in the fa/fa rats fed the high-fat diet as lipid peroxidation and protein carbonyls were elevated, while glutathione and antioxidant enzymes were very low. Expression and activity of cytochrome P450 2E1 and xanthine oxidase activity were down-regulated in fa/fa compared with Fa/? rats, and no effect was seen by the high-fat diet. However, NADPH oxidase activity increased 2.5-fold in fa/fa rats fed with the high-fat diet. In summary, a high-fat diet induces liver injury in fa/fa rats leading to periportal fibrosis. A role for oxidative stress is suggested via increased NADPH oxidase activity, lipid peroxidation, protein carbonyl formation, and low antioxidant defense. PMID- 15522906 TI - Dietary lipid hydroperoxides induce expression of vascular endothelial growth factor (VEGF) in human colorectal tumor cells. AB - Fatty acid hydroperoxides arise from unsaturated fatty acids in the presence of oxygen and elevated temperature during processing of food. Here we have studied their effects on gene expression in colorectal tumor cells using linoleic acid hydroperoxide (LOOH) as a model compound. Its addition to the medium of LT97 human adenoma cells and SW480 human carcinoma cells enhanced the production of intracellular hydrogen peroxide. Furthermore, in both cell lines, increases in VEGF mRNA and protein were observed. Unoxidized linoleic acid had little or no activity. Concomitantly, COX-2 expression was up-regulated. In the LT97 cells, the COX inhibitors SC58560 and SC58236 completely prevented the VEGF induction, suggesting that the effect was dependent on prostaglandin synthesis. In vivo prostaglandin-mediated induction of VEGF secretion is known to be essential for the growth of adenomatous polyps and their progression to carcinomas. Therefore, our results for the first time implicate dietary lipid hydroperoxide as a key risk factor in colon carcinogenesis. PMID- 15522907 TI - Neuronal plasticity after spinal cord injury: identification of a gene cluster driving neurite outgrowth. AB - Functional recovery after spinal cord injury (SCI) may result in part from axon outgrowth and related plasticity through coordinated changes at the molecular level. We employed microarray analysis to identify a subset of genes the expression patterns of which were temporally coregulated and correlated to functional recovery after SCI. Steady-state mRNA levels of this synchronously regulated gene cluster were depressed in both ventral and dorsal horn neurons within 24 h after injury, followed by strong re-induction during the following 2 wk, which paralleled functional recovery. The identified cluster includes neuritin, attractin, microtubule-associated protein 1a, and myelin oligodendrocyte protein genes. Transcriptional and protein regulation of this novel gene cluster was also evaluated in spinal cord tissue and in single neurons and was shown to play a role in axonal plasticity. Finally, in vitro transfection experiments in primary dorsal root ganglion cells showed that cluster members act synergistically to drive neurite outgrowth. PMID- 15522908 TI - Inhibition of host connective tissue growth factor expression: a novel Trypanosoma cruzi-mediated response. AB - Connective tissue growth factor (CTGF) is a secreted cytokine that plays a fundamental role in the development of tissue fibrosis by mediating many of the profibrotic effects of TGF-beta. We present the novel finding that the intracellular pathogen Trypanosoma cruzi elicits immediate and sustained repression of basal CTGF expression in dermal fibroblasts, followed by down regulation of the extracellular matrix proteins, fibronectin, and collagen I alpha1. To address mechanisms underlying this response, the major CTGF-regulating pathways were investigated. We report that both T. cruzi trypomastigotes and secreted parasite factor(s) antagonize TGF-beta-dependent induction of CTGF in fibroblasts. Of the TGF-beta-dependent signaling pathways required for CTGF expression, we demonstrate that T. cruzi interferes with cellular Erk1/2 phosphorylation but not Smad3 signaling. While increased stimulation of Erk phosphorylation alone was insufficient to override the parasite-mediated repression of CTGF, stimulation of fibroblasts with increased concentrations of TGF-beta, which activates both Smad3 and Erk1/2, completely abrogated this inhibition. Together with the finding that T. cruzi-mediated down-regulation of CTGF expression requires de novo host cell protein synthesis, our data indicate that the unique ability of T. cruzi to interfere with the host fibrogenic response is a complex process requiring input from multiple host cell signaling pathways. PMID- 15522909 TI - Hsc70 and Hsp70 interact with phosphatidylserine on the surface of PC12 cells resulting in a decrease of viability. AB - Heat shock proteins (hsps) are involved in multiple cellular processes during normal and stress conditions, particularly in the folding of polypeptides. A newly recognized property of the members of the Hsp70 family is their ability to interact with lipids, opening ion conductance pathways in artificial membranes, and integrating into natural membranes. The formation of Hsp70 channels in biological membranes and their function is still elusive. In this study, we showed that Hsp70 and Hsc70 display a highly selective interaction with phosphatidylserine moieties on membranes, followed by rapid incorporation into the lipid bilayer. Addition of Hsp70 or Hsc70 into the extracellular medium resulted in a viability decrease of cells beading PS on the exterior surface, such as PC12 cells. This toxic effect is modulated by the presence of ATP or ADP and can be blocked by screening PS moieties with annexin 5. These observations suggest that the presence of Hsp70 in the extracellular medium may be an accelerator of apoptosis since the presence of PS on the surface is an early indicator of this process. These findings may also explain the toxicity observed in cells overexpressing Hsp70s and provide a rational for the tight regulation of Hsp70 expression. PMID- 15522910 TI - Melatonin desensitizes endogenous MT2 melatonin receptors in the rat suprachiasmatic nucleus: relevance for defining the periods of sensitivity of the mammalian circadian clock to melatonin. AB - The hormone melatonin phase shifts circadian rhythms generated by the mammalian biological clock, the suprachiasmatic nucleus (SCN) of the hypothalamus, through activation of G protein-coupled MT2 melatonin receptors. This study demonstrated that pretreatment with physiological concentrations of melatonin (30-300 pM or 7 70 pg/mL) decreased the number of hMT2 melatonin receptors heterologously expressed in mammalian cells in a time and concentration-dependent manner. Furthermore, hMT2-GFP melatonin receptors heterologously expressed in immortalized SCN2.2 cells or in non-neuronal mammalian cells were internalized upon pretreatment with both physiological (300 pM or 70 pg/mL) and supraphysiological (10 nM or 2.3 ng/mL) concentrations of melatonin. The decrease in MT2 melatonin receptor number induced by melatonin (300 pM for 1 h) was reversible and reached almost full recovery after 8 h; however, after treatment with 10 nM melatonin full recovery was not attained even after 24 h. This recovery process was partially protein synthesis dependent. Furthermore, exposure to physiological concentrations of melatonin (300 pM) for a time mimicking the nocturnal surge (8 h) desensitized functional responses mediated through melatonin activation of endogenous MT2 receptors, i.e., stimulation of protein kinase C (PKC) in immortalized SCN2.2 cells and phase shifts of circadian rhythms of neuronal firing in the rat SCN brain slice. We conclude that in vivo the nightly secretion of melatonin desensitizes endogenous MT2 melatonin receptors in the mammalian SCN thereby providing a temporally integrated profile of sensitivity of the mammalian biological clock to a melatonin signal. PMID- 15522911 TI - Weight loss regulates inflammation-related genes in white adipose tissue of obese subjects. AB - Adipose tissue produces inflammation and immunity molecules suspected to be involved in obesity-related complications. The pattern of expression and the nutritional regulation of these molecules in humans are poorly understood. We analyzed the gene expression profiles of subcutaneous white adipose tissue from 29 obese subjects during very low calorie diet (VLCD) using cDNA microarray and reverse transcription quantitative PCR. The patterns of expression were compared with that of 17 non-obese subjects. We determined whether the regulated genes were expressed in adipocytes or stromavascular fraction cells. Gene expression profiling identified 100 inflammation-related transcripts that are regulated in obese individuals when eating a 28 day VLCD but not a 2 day VLCD. Cluster analysis showed that the pattern of gene expression in obese subjects after 28 day VLCD was closer to the profile of lean subjects than to the pattern of obese subjects before VLCD. Weight loss improves the inflammatory profile of obese subjects through a decrease of proinflammatory factors and an increase of anti inflammatory molecules. The genes are expressed mostly in the stromavascular fraction of adipose tissue, which is shown to contain numerous macrophages. The beneficial effect of weight loss on obesity-related complications may be associated with the modification of the inflammatory profile in adipose tissue. PMID- 15522912 TI - Novel and therapeutic effect of caffeic acid and caffeic acid phenyl ester on hepatocarcinoma cells: complete regression of hepatoma growth and metastasis by dual mechanism. AB - Our previous studies have clearly shown that the angiogenic enzymes, matrix metalloproteinase (MMP) -2/9, are directly involved in human hepatic tumorigenesis and metastasis and suggest that the MMP-2/9 inhibitors, which have dual inhibitory activities on enzyme activity and transcription, represent the best candidates for achieving tumor regression. Many anti-cancer drugs have strong cellular cytotoxicity and side effects, indicating that strong anti-cancer drugs that have no or minimal cytotoxicity and side effects need to be developed. The specific aim of the present study was to develop powerful anti-cancer drugs with specific tumor regression and anti-metastatic potential having the dual inhibitory activities of specific MMP-2 and -9 enzyme activities and gene transcription at the molecular level. Caffeic acid (CA), a strong and selective MMP-9 activity and transcription inhibitor, was isolated from the plant Euonymus alatus and its derivative, caffeic acid phenethyl ester (CAPE), was synthesized. CA and CAPE selectively inhibited MMP-2 and -9 but not -1, -3, -7, or cathepsin K. Treatment of HepG2 cells with CA (100 microg/mL) and CAPE (5 microg/mL) suppressed phorbol 12-myristate 13-acetate (PMA) -induced MMP-9 expression by inhibiting the function of NF-kappaB, but not AP-1. We confirmed that CA and CAPE suppressed the growth of HepG2 tumor xenografts in nude mice in vivo. The subcutaneous and oral administrations of CA and CAPE significantly reduced the liver metastasis. These results confirm the therapeutic potential of the compounds and suggest that the anti-metastatic and anti-tumor effects of CA and CAPE are mediated through the selective suppression of MMP-9 enzyme activity and transcriptional down-regulation by the dual inhibition of NF-kappaB as well as MMP-9 catalytic activity. PMID- 15522913 TI - MMP-2 null mice exhibit an early onset and severe experimental autoimmune encephalomyelitis due to an increase in MMP-9 expression and activity. AB - Matrix metalloproteinase-2 (MMP-2; gelatinase A) is known to degrade a broad range of extracellular matrix components and chemokines, and has important roles in the processes of cell migration, invasion, and involution during development, as well as during tumor growth and metastasis and in inflammation and repair. To better elucidate the roles of this matrix metalloproteinase in the development and progression of experimental autoimmune encephalomyelitis, we used MMP-2 deficient (KO) mice. Surprisingly, we found that MMP-2 KO mice exhibited an earlier onset and more severe disease than did their wild-type (WT) counterparts. WT mice engrafted with MMP-2 KO bone marrow exhibited a similar earlier onset and more severe clinical disease score than WT mice engrafted with WT bone marrow. Lymphocytes derived from MMP-2 KO mice exhibited increased transmigration through endothelial cell monolayers as well as through collagen type IV and laminin coated BD BIOCOAT inserts, which correlated with a 3-fold increase in expression of MMP-9 and was abrogated by inhibition of MMP activity. We demonstrated a correlation between expression levels of MMP-9 and MT1-MMP expression and suggest a signaling pathway involving tethering of MMP-2 to MT1-MMP as a modulator of MMP 9 expression. Last, we discuss other possible MMP-2-mediated mechanisms which may contribute to the observed phenotype. PMID- 15522914 TI - Intramyocardial lipid accumulation in the failing human heart resembles the lipotoxic rat heart. AB - In animal models of lipotoxicity, accumulation of triglycerides within cardiomyocytes is associated with contractile dysfunction. However, whether intramyocardial lipid deposition is a feature of human heart failure remains to be established. We hypothesized that intramyocardial lipid accumulation is a common feature of non-ischemic heart failure and is associated with changes in gene expression similar to those found in an animal model of lipotoxicity. Intramyocardial lipid staining with oil red O and gene expression analysis was performed on heart tissue from 27 patients (9 female) with non-ischemic heart failure. We determined intramyocardial lipid, gene expression, and contractile function in hearts from 6 Zucker diabetic fatty (ZDF) and 6 Zucker lean (ZL) rats. Intramyocardial lipid overload was present in 30% of non-ischemic failing hearts. The highest levels of lipid staining were observed in patients with diabetes and obesity (BMI>30). Intramyocardial lipid deposition was associated with an up-regulation of peroxisome proliferator-activated receptor alpha (PPARalpha) -regulated genes, myosin heavy chain beta (MHC-beta), and tumor necrosis factor alpha (TNF-alpha). Intramyocardial lipid overload in the hearts of ZDF rats was associated with contractile dysfunction and changes in gene expression similar to changes found in failing human hearts with lipid overload. Our findings identify a subgroup of patients with heart failure and severe metabolic dysregulation characterized by intramyocardial triglyceride overload and changes in gene expression that are associated with contractile dysfunction. PMID- 15522915 TI - Single-chain antibodies for the conformation-specific blockade of activated platelet integrin alphaIIbbeta3 designed by subtractive selection from naive human phage libraries. AB - Binding of fibrinogen to platelet integrin alphaIIbbeta3 mediates platelet aggregation, and thus inhibition of alphaIIbbeta3 represents a powerful therapeutic strategy in cardiovascular medicine. However, the currently used inhibitors of alphaIIbbeta3 demonstrate several adverse effects like thrombocytopenia and bleeding, which are associated with their property to bind to non-activated alphaIIbbeta3. To circumvent these problems, we designed blocking single-chain antibody-fragments (scFv) that bind to alphaIIbbeta3 exclusively in its activated conformation. Two naive phage libraries were created: a natural phage library, based on human lymphocyte cDNA, and a synthetic library, with randomized VHCDR3. We performed serial rounds of subtractive panning with depletion on non-activated and selection on activated alphaIIbbeta3, which were provided on resting and ADP-stimulated platelets and CHO cells, expressing wild-type or mutated and thereby activated alphaIIbbeta3. In contrast to isolated, immobilized targets, as generally used for phage display, this unique cell-based approach for panning allowed the preservation of functional integrin conformation. Thereby, we obtained several scFv-clones that demonstrated exclusive binding to activated platelets and complete inhibition of fibrinogen binding and platelet aggregation. Interestingly, all activation-specific clones contained an RXD pattern in the HCDR3. Binding studies on transiently expressed point mutants and mouse-human domain-switch mutants of alphaIIbbeta3 indicate a binding site similar to fibrinogen. In conclusion, we generated human activation specific scFvs against alphaIIbbeta3, which bind selectively to activated alphaIIbbeta3 and thereby potently inhibit fibrinogen binding to alphaIIbbeta3 and platelet aggregation. PMID- 15522916 TI - New insights into the molecular mechanism of interleukin-10-mediated immunosuppression. AB - Interleukin-10 (IL-10) is an important immunomodulatory cytokine, which has attracted much attention because of its anti-inflammatory properties. It reduces antigen presentation and inhibits T cell activation. IL-10-treated myeloid cells lose their ability to respond toward the endotoxin lipopolysaccharide (LPS) with the production of several proinflammatory mediators. Thereby, IL-10 limits excessive inflammatory reactions in response to endotoxin as it occurs in colitis or endotoxin shock. Mice can be tolerized toward endotoxin shock when pretreated with a sublethal dose of LPS. This can be mimicked in vitro as LPS desensitization, resulting in a similar LPS hyporesponsiveness as observed with IL-10 pretreatment. However, an early block in LPS signaling characterizes LPS desensitization, whereas IL-10 seems to target late events. Controversial reports have been published where IL-10 would interfere with the induction of proinflammatory mediators, and little is known about the molecular mechanisms behind the anti-inflammatory activities of IL-10. Some recent publications have tried to gain more insight into the molecular mechanism of IL-10 by gene expression profiling and functional studies in myeloid-derived cells. These results are reviewed here and compared with the progress that has been made to understand the induction of endotoxin tolerance by LPS itself. PMID- 15522917 TI - The CD4+ T cell immunodominant Anaplasma marginale major surface protein 2 stimulates gammadelta T cell clones that express unique T cell receptors. AB - Major surface protein 2 (MSP2) of the bovine rickettsial pathogen Anaplasma marginale is an abundant, serologically immunodominant outer membrane protein. Immunodominance partially results from numerous CD4+ T cell epitopes in highly conserved amino and carboxy regions and the central hypervariable region of MSP2. However, in long-term cultures of lymphocytes stimulated with A. marginale, workshop cluster 1 (WC1)+ gammadelta T cells and CD4+ alphabeta T cells proliferated, leading to a predominance of gammadelta T cells. As gammadelta T cells proliferate in A. marginale-stimulated lymphocyte cultures, this study hypothesized that gammadelta T cells respond to the abundant, immunodominant MSP2. To test this hypothesis, gammadelta T cell clones were isolated from MSP2 vaccinates and assessed for antigen-specific proliferation and interferon-gamma secretion. Seven WC1+ gammadelta T cell clones responded to A. marginale and MSP2, and three of these proliferated to overlapping peptides from the conserved carboxy region. The gammadelta T cell response was not major histocompatibility complex-restricted, although it required antigen-presenting cells and was blocked by addition of antibody specific for the T cell receptor (TCR). Sequence analysis of TCR-gamma and -delta chains of peripheral blood lymphocytes identified two novel TCR-gamma chain constant (Cgamma) regions. It is important that all seven MSP2-specific gammadelta T cell clones used the same one of these novel Cgamma regions. The TCR complementarity-determining region 3 was less conserved than those of MSP2-specific CD4+ alphabeta T cell clones. Together, these data indicate that WC1+ gammadelta T cells recognize A. marginale MSP2 through the TCR and contribute to the immunodominant response to this protein. PMID- 15522918 TI - Lysophosphatidic acid triggers calcium entry through a non-store-operated pathway in human neutrophils. AB - Lysophosphatidic acid (LPA) is a bioactive lipid, which is structurally similar to sphingosine 1-phosphate (S1P) and which can mobilize Ca2+ in multiple cell types. We recently showed that S1P induces Ca2+ entry directly through store operated Ca2+ entry (SOCE) channels in human polymorphonuclear neutrophils (PMN). We therefore examined the mechanisms by which LPA induces intracellular Ca2+ mobilization in PMN. External application of low micromolar LPA caused dose dependent Ca2+ influx without releasing Ca2+ stores, whereas G-protein-coupled (GPC) LPA receptors respond to nanomolar LPA. Additive Ca2+ influx by LPA compared with 100 nM ionomycin-induced Ca2+ influx suggests that LPA-induced Ca2+ influx does not pass through SOCE channels. Ca2+ influx was resistant to inhibition of Gi/o by pertussis toxin, of phospholipase C by U73122, and of G12/13/Rho by Y27632, all demonstrating GPC receptor independence. This Ca2+ influx was inhibited by Gd3+, La3+, Zn2+, or MRS1845 but not by Ni2+ or the sphingosine kinase inhibitor dimethylsphingosine. In addition, we found that LPA has no effect on neutrophil chemotaxis; however, it has stimulatory effects on neutrophil respiratory burst in a dose-response manner. These findings suggest that LPA-induced Ca2+ influx in PMN occurs through a mechanism other than SOCE channels, independent of Ca2+ store-depletion and S1P synthesis, and that the characteristics of LPA-induced Ca2+ influx are similar to those of S1P-induced influx in terms of sensitivity to inorganic inhibitors. Unlike S1P, LPA has stimulatory effects on neutrophil respiratory burst. PMID- 15522919 TI - Human cytomegalovirus inhibits the migration of immature dendritic cells by down regulating cell-surface CCR1 and CCR5. AB - Dendritic cells (DC) play a key role in the host immune response to infections. Human cytomegalovirus (HCMV) infection can inhibit the maturation of DC and impair their ability to stimulate T cell proliferation and cytotoxicity. In this study, we assessed the effects of HCMV infection on the migratory behavior of human DC. The HCMV strain TB40/E inhibited the migration of immature monocyte derived DC in response to inflammatory chemokines by 95% 1 day after infection. This inhibition was mediated by early viral replicative events, which significantly reduced the cell-surface expression of CC chemokine receptor 1 (CCR1) and CCR5 by receptor internalization. HCMV infection also induced secretion of the inflammatory chemokines CC chemokine ligand 3 (CCL3)/macrophage inflammatory protein-1alpha (MIP-1alpha), CCL4/MIP-1beta, and CCL5/regulated on activation, normal T expressed and secreted (RANTES). Neutralizing antibodies for these chemokines reduced the effects of HCMV on chemokine receptor expression and on DC migration by approximately 60%. Interestingly, the surface expression of the lymphoid chemokine receptor CCR7 was not up-regulated after HCMV infection on immature DC, and immature-infected DC did not migrate in response to CCL19/MIP 3beta. These findings suggest that blocking the migratory ability of DC may be a potent mechanism used by HCMV to paralyze the early immune response of the host. PMID- 15522920 TI - Covalent DNA display as a novel tool for directed evolution of proteins in vitro. AB - We present a novel method for the directed evolution of polypeptides, which combines in vitro compartmentalization and covalent DNA display. A library of linear DNA fragments is co-packaged with an in vitro transcription/translation mixture in the compartments of a water-in-oil emulsion. Experimental conditions are adjusted so that, in most cases, one compartment contains one DNA molecule. The DNA fragments encode fusion proteins containing a DNA-methyltransferase (M.Hae III), which can form a covalent bond with a 5-fluorodeoxycytidine base at the extremity of the DNA fragment. The resulting library of DNA-protein fusions is extracted from the emulsion and DNA molecules displaying a protein with desired binding properties are selected from the pool of DNA-protein fusions by affinity panning on target antigens. We applied this methodology in model selection experiments, using specific ligands for the capture of peptides and globular proteins bound to DNA. We observed enrichment factors >1000-fold for selections performed in separate emulsions and up to 150-fold for selections performed using mixtures of DNA molecules. M.Hae III could be fused to small globular proteins (such as calmodulin and fibronectin domains), which are ideally suited for the generation of combinatorial libraries and for the isolation of novel binding specificities. PMID- 15522921 TI - Contribution of MHC class I chain-related A (MICA) gene polymorphism to genetic susceptibility for systemic lupus erythematosus. AB - OBJECTIVE: To evaluate the contribution of the MHC class I chain-related A (MICA) gene polymorphism to the genetic risk of systemic lupus erythematosus (SLE). METHODS: HLA-DRB1-DQA1-DQB1 genotyping, MICA exon 5 microsatellite genotyping and HLA-B8 genotyping were performed in 48 Italian SLE patients and in 158 healthy control subjects. RESULTS: Of HLA class II haplotypes, only DRB1*03-DQA1*0501 DQB1*0201 (DR3-DQ2) was significantly more frequent among SLE patients than among healthy control subjects [odds ratio (OR) = 6.5, corrected P < 0.0026]. HLA-B8 was detected in 31% SLE patients and 13% healthy control subjects (OR = 3.0, P = 0.005). The allele-wise comparison between patients and controls showed that both MICA5 (OR = 2.5, corrected P < 0.0005) and MICA5.1 (OR = 2.4, corrected P < 0.0005) were positively and MICA9 (OR = 0.2, corrected P < 0.0005) was negatively associated with the disease. The MICA5/5.1 genotype was positively associated with SLE (OR = 28.9, corrected P < 0.0015) also in subjects negative for DR3-DQ2 (OR > 22.6, corrected P < 0.011). The simultaneous presence of DR3-DQ2 and MICA5.1 was detected in 15/48 (31%) SLE and in 10/158 (6%) healthy control subjects (OR = 6.7, corrected P < 0.011). The simultaneous combination of DR3-DQ2 and MICA5 was found in 10/48 (21%) SLE patients and in only 1/158 healthy control subjects (OR = 41.3, corrected P < 0.011). Logistic regression analysis showed the independent positive associations of MICA5 and MICA5.1 and negative association of MICA9 with the disease, and revealed that the interaction of the three major markers (DR3-DQ2, MICA5 and MICA5.1) was associated with increasing genetic risk, which was highest (OR > 30.3) in DR3-DQ2-positive subjects carrying the MICA5-5.1 genotype. CONCLUSIONS: Our study provides the first demonstration of the independent association of the MICA gene polymorphism with genetic risk of SLE. PMID- 15522922 TI - Cardiovascular disease and psychological morbidity among rheumatoid arthritis patients. AB - OBJECTIVES: To examine whether patients with rheumatoid arthritis (RA) with co morbid cardiovascular disease (CVD) have different psychological morbidity (and psychosocial risk factors for it) compared with RA patients without co-morbid CVD. METHODS: Patients with RA and co-morbid CVD (n = 44; hypertension alone for n = 27) were compared with RA patients without CVD (n = 110). Differences in psychological morbidity (depression and anxiety) and psychosocial risk factors for this (arthritis self-efficacy, acceptance, social support and optimism) were examined while controlling statistically for medical and demographic covariates. RESULTS: Groups did not differ on RA duration, RA activity, marital status or socioeconomic status, but RA patients with co-morbid CVD were older, less likely to be female and less likely to be in employment than those without CVD. RA patients with co-morbid CVD had significantly higher depression and were more likely to score above cut-offs for depression than RA patients without CVD. No differences existed in anxiety, although anxiety appeared to be more common than depression. Low optimism was identified as a possible psychosocial risk factor for depression. CONCLUSIONS: RA patients with co-morbid CVD have higher depression than RA patients without CVD; low optimism is a potentially modifiable risk factor that may mediate this difference. RA patients with co-morbid CVD may benefit from systematic screening for depression and targeted intervention if necessary. PMID- 15522923 TI - Commentary: The age distribution of cancer and a multistage theory of carcinogenesis. PMID- 15522924 TI - Risk factors for prematurity at Harare Maternity Hospital, Zimbabwe. AB - BACKGROUND: Prematurity remains the main cause of mortality and morbidity in infants and a problem in the care of pregnant women world-wide. This preliminary study describes the socio-demographic, reproductive, medical, and obstetrical risk factors for having a live pre-term delivery (PTD) in Zimbabwe. METHODS: This case-control study examined risk factors for PTD, at Harare Maternity Hospital between March and June 1999. RESULTS: The frequency of PTD among live birth was 16.4%. Prior history of stillbirth or abortion was associated with PTD (adjusted relative risk [ARR] 1.50; 95% CI: 1.06, 2.11). Nutritional factors, including drinking a local non-alcoholic beverage (mahewu) during pregnancy and mother's increasing mid-arm circumference reduced the risk of PTD (ARR = 0.75; 95% CI: 0.60, 0.93 and ARR = 0.95; 95% CI: 0.92, 0.99 per cm of circumference, respectively). Obstetric conditions including eclampsia, anaemia, ante-partum haemorrhage, and placenta praevia were infrequent, but when present, were strongly associated with PTD (ARR = 3.57; 95% CI: 1.67, 7.63; ARR = 4.12; 95% CI: 1.80, 9.43; ARR = 3.05; 95% CI: 1.86, 5.00 and ARR = 3.30; 95% CI: 1.34, 8.14, respectively). Malaria, although less frequent, nonetheless was associated with an increased risk of PTD (ARR = 2.93; 95% CI: 1.70, 5.04). These results suggest that in addition to established obstetric risk factors, nutrition and malarial infection are important. About 43% of the mothers initiated prenatal care after 28 weeks of gestation. CONCLUSION: Addressing prematurity in this population will require earlier initiation of prenatal care to allow for early detection and management of complications of pregnancy, and improving nutritional status of reproductive age with locally available foods. Further exploration of the potential benefits of mahewu, is warranted. PMID- 15522926 TI - The association of thyroid function with cardiac mass and left ventricular hypertrophy. AB - Decreased serum TSH levels predict cardiovascular mortality, which could be explained by left ventricular hypertrophy (LVH). The aim of this analysis was to investigate the association between thyroid function and LVH. The population based Study of Health in Pomerania was conducted in a previously iodine-deficient area. Data of 1510 individuals at least 45 yr of age with echocardiography and without thyroid disorders were analyzed. LVH was defined as a left ventricular mass index (LVMI) exceeding 150 g/m(2) (men) or 120 g/m(2) (women). Overt hyperthyroidism was associated with LVMI (P < 0.01), whereas euthyroid subjects and those with elevated TSH levels did not significantly differ with regard to LVMI. LVH was observed in three (15.0%) subjects with elevated serum TSH levels, in 127 (10.5%) euthyroid persons, in 24 (12.5%) individuals with decreased serum TSH levels, and in four (57.1%) subjects with hyperthyroidism (P < 0.01). Logistic regression analysis identified overt hyperthyroidism as an independent risk factor for LVH (odds ratio, 13.65; 95% confidence interval, 2.83-65.75; P < 0.01). There is an association between thyroid function status, cardiac mass, and LVH. Hyperthyroidism is an independent risk factor for LVH. PMID- 15522925 TI - Delta-4-androstene-3,17-dione binds androgen receptor, promotes myogenesis in vitro, and increases serum testosterone levels, fat-free mass, and muscle strength in hypogonadal men. AB - Previous studies of Delta 4-androstene-3,17-dione (4-androstenedione) administration in men have not demonstrated sustained increments in testosterone levels, fat-free mass (FFM), and muscle strength, and failure to demonstrate androstenedione's androgenic/anabolic effects has stifled efforts to regulate its sales. To determine whether 4-androstenedione has androgenic/anabolic properties, we evaluated its association with androgen receptor (AR) and its effects on myogenesis in vitro. Additionally, we studied the effects of a high dose of 4 androstenedione on testosterone levels, FFM, and muscle strength in hypogonadal men. We determined the dissociation constant (K(d)) for 4-androstenedione using fluorescence anisotropy measurement of competitive displacement of fluorescent androgen from AR ligand-binding domain. AR nuclear translocation and myogenic activity of androstenedione were evaluated in mesenchymal, pluripotent C3H10T1/2 cells, in which androgens stimulate myogenesis through an AR pathway. We determined effects of a high dose of androstenedione (500 mg thrice daily) given for 12 wk on FFM, muscle strength, and hormone levels in nine healthy, hypogonadal men. 4-Androstenedione competitively displaced fluorescent androgen from AR ligand-binding domain with a lower affinity than dihydrotestosterone (K(d), 648 +/- 21 and 10 +/- 0.4 nm, respectively). In C3H10T1/2 cells, 4 androstenedione caused nuclear translocation of AR and stimulated myogenesis, as indicated by a dose-dependent increase in myosin heavy chain II+ myotube area and up-regulation of MyoD protein. Stimulatory effects of 4-androstenedione on myosin heavy chain II+ myotubes and myogenic determination factor expression were attenuated by bicalutamide, an AR antagonist. Administration of 1500 mg 4 androstenedione daily to hypogonadal men significantly increased serum androstenedione, total and free testosterone, estradiol, and estrone levels and suppressed SHBG and high-density lipoprotein cholesterol levels. 4 androstenedione administration was associated with significant gains in FFM (+1.7 +/- 0.5 kg; P = 0.012) and muscle strength in bench press (+4.3 +/- 3.1 kg; P = 0.006) and leg press exercises (+18.8 +/- 17.3 kg; P = 0.045). 4-androstenedione is an androgen that binds AR, induces AR nuclear translocation, and promotes myogenesis in vitro, with substantially lower potency than dihydrotestosterone. 4 androstenedione administration in high doses to hypogonadal men increases testosterone levels, FFM, and muscle strength, although at the dose tested, the anabolic effects in hypogonadal men are likely because of its conversion to testosterone. PMID- 15522927 TI - Beneficial postprandial effect of a small amount of alcohol on diabetes and cardiovascular risk factors: modification by insulin resistance. AB - Moderate alcohol consumption protects against type 2 diabetes and cardiovascular disease. Because humans spend most of their time in the postprandial state, we examined the effect of 15 g alcohol on postprandial metabolic factors in 20 postmenopausal women over 6 h. We measured 1) glucose, insulin, lipids, C reactive protein, and adiponectin levels; 2) augmentation index by applanation tonometry; and 3) energy expenditure and substrate oxidation by indirect calorimetry. Subjects received low carbohydrate (LC; visits 1 and 2) and high carbohydrate (HC; visits 3 and 4) high fat meals with and without alcohol. Alcohol augmented the postprandial increment in insulin (P = 0.07) and reduced the postprandial increment in glucose (P = 0.04) after the LC meal only. Triglycerides were increased by alcohol after the LC (P = 0.002) and HC (P = 0.008) meals. Total and high-density lipoprotein cholesterol, fatty acids, and total adiponectin responses were unaffected. C-reactive protein levels decreased postprandially; reductions were enhanced by alcohol after the HC meal, but were attenuated after the LC meal. Postprandial reductions in the augmentation index were increased by alcohol after the LC meal only (P = 0.007). Alcohol enhanced the postprandial increase in energy expenditure 30-60 min after the LC meal (increase, 373 +/- 49 vs. 236 +/- 32 kcal/d; P = 0.02) and HC meal (increase, 362 +/- 36 vs. 205 +/- 34 kcal/d; P = 0.0009), but suppressed fat and carbohydrate oxidation. Some of our findings may be mechanisms for lower diabetes and cardiovascular risks in moderate drinkers. PMID- 15522928 TI - Adult height after ketoconazole treatment in patients with familial male-limited precocious puberty. AB - Familial male-limited precocious puberty is a rare cause of precocious puberty due to activating mutations of the LH receptor, leading to early onset virilization and short stature. Two therapeutic approaches have been proposed: the P450 cytochrome inhibitor ketoconazole or combined treatment with spironolactone and testolactone. Results on adult heights have not been reported to date after these two treatments, and in this study we present results from five patients treated with ketoconazole at a median dose of 16.2 mg/kg.d for a median of 6.2 yr. Adult height was 173 cm (median; interquartile range, 14), similar to target height (175 cm; interquartile range, 9) and significantly higher than pretreatment predicted height (165 cm; interquartile range, 12; P < 0.01). During treatment, 39 of 58 (68%) testosterone measurements were less than 0.5 ng/ml (1.7 nmol/liter), nine of 58 (15%) were between 0.5 and 1 ng/ml (3.5 nmol/liter), and 10 of 58 (17%) were above 1 ng/ml. We observed a physiological increase in GnRH-stimulated LH levels after the age of 10 yr, and none of the patients had early activation of the gonadotropic axis. Liver tolerance was excellent, and only one patient had a transient and modest increase in serum transaminases. We conclude that ketoconazole is an efficient and well tolerated long-term treatment of familial male-limited precocious puberty that should be proposed as a first line therapy. PMID- 15522929 TI - Gonadotroph tumor associated with multiple endocrine neoplasia type 1. AB - Although anterior pituitary tumors constitute a main clinical feature of multiple endocrine neoplasia type 1 (MEN1), and most types of pituitary tumors have been associated with MEN1, gonadotroph tumors have not previously been recognized clinically as part of this syndrome. We report here a woman who presented with ovarian hyperstimulation due to a gonadotroph tumor that was confirmed biochemically and immunohistochemically. She then developed hyperparathyroidism, and she was found to have three hypercellular parathyroid glands. Subsequently, she developed a temporal lobe metastasis of the gonadotroph tumor, demonstrating that it was a gonadotroph carcinoma. The diagnosis of MEN1 was confirmed by finding a deletion mutation (c.307delC) on the second exon of the MEN1 gene that predicts truncation of the resulting menin protein 15 codons downstream from the deletion (p.Leu103fsX15). This case illustrates that gonadotroph tumors, like other pituitary tumors, can be part of MEN1. The clinical implications of this case are that the clinical and biochemical features of gonadotroph tumors should be considered when evaluating patients for MEN1, and MEN1 should be considered in patients who have gonadotroph tumors. PMID- 15522930 TI - Essential role of fibroblast growth factor signaling in preadipoctye differentiation. AB - We have examined the expression and role of autocrine fibroblast growth factors (FGFs) in human preadipocytes through their differentiation in vitro. A high molecular weight form of FGF-2 was initially strongly expressed, but 6-9 d after induction of differentiation, its expression decreased markedly. This coincided with the first appearance of visible lipid droplets within the cells. FGF-2 (18 kDa) was not found. FGF receptor (FGFR) 1 was detected as a single band of 125 kDa that also decreased with differentiation. Its decrease preceded that of FGF 2. Despite the decrease in cell-associated FGF-2 with differentiation, secreted FGF-2 was 2.5-fold higher in the differentiated preadipocytes. To determine whether FGF-2 had an autocrine role, FGFR signaling was inhibited using recombinant adenovirus expressing dominant negative FGFR1 (RAdDN-FGFR1) and a specific inhibitor of FGFR1 signaling, PD166866. Preadipocytes transduced with RAdDN-FGFR1 expressed a truncated, 79-kDa FGFR1. Differentiation, assessed by lipid droplet formation, was completely prevented by RAdDN-FGFR1 and by PD166866. The protein content in the cell layer and glucose uptake were significantly reduced by both agents. The insulin-sensitizing drug, rosiglitazone, did not prevent the actions of RAdDN-FGFR1 or PD166866. Controlling adipose tissue growth by limiting FGF actions may provide a means to combat obesity. PMID- 15522931 TI - Antiretroviral treatment reduces very-low-density lipoprotein and intermediate density lipoprotein apolipoprotein B fractional catabolic rate in human immunodeficiency virus-infected patients with mild dyslipidemia. AB - The relationship between antiretroviral treatment of HIV infection, body fat distribution, insulin resistance, and very-low-density lipoprotein (VLDL) and intermediate-density lipoprotein (IDL) apolipoprotein-B (apoB) kinetics was investigated in 55 HIV-infected patients taking two nucleoside analogs plus either a protease inhibitor (n = 15) or a nonnucleoside reverse transcriptase inhibitor (n = 25), 15 antiretroviral therapy-naive patients, and 12 HIV-negative controls. Compared with the controls, high-density lipoprotein cholesterol was reduced in all groups (P < 0.01). Plasma triglyceride was increased in patients taking protease inhibitors (P < 0.05). VLDL and IDL apoB fractional catabolic rate (FCR) was lower in all treatment groups (P < 0.05) compared with controls. Trunk fat, VLDL apoB absolute secretion rate, and insulin resistance were not different between groups. Peripheral fat was lower in the treated patients (P < 0.05) and correlated with duration of therapy (r = -0.55; P < 0.001). There was a positive correlation between peripheral fat and VLDL apoB FCR (P = 0.002) and IDL apoB FCR (P = 0.002) and a negative correlation with VLDL apoB pool size, VLDL cholesterol, and triglyceride (P < 0.03; P < 0.01; P < 0.002). These results suggest that mild dyslipidemia resulting from antiretroviral therapy is caused by a decrease in VLDL and IDL apoB FCR, which is associated with a loss of peripheral fat. PMID- 15522932 TI - The normal glucose tolerance continuum in obese youth: evidence for impairment in beta-cell function independent of insulin resistance. AB - Normal glucose tolerance is expressed over a wide range of glucose concentrations. Whether there is a continuum of risk for developing type 2 diabetes mellitus even when the 2-h plasma glucose is still within this normal range is uncertain. Oral glucose tolerance tests were performed in 407 obese normal glucose tolerance youth (4-20 yr) to examine the relationship between variations in 2-h plasma glucose levels and beta-cell responsiveness. Individuals were grouped by 2-h plasma glucose levels as follows: 1) less than 100 mg/dl, 2) 100-119 mg/dl, and 3) 120-139 mg/dl. Subsequent analysis stratified each 2-h plasma level by insulin sensitivity index. Increased 2-h glucose level was associated with a progressive increase in glucose between 0 and 30 min (P < 0.05). The Delta (0-30 min) insulin did not vary significantly across levels, thus resulting in a decreased insulinogenic index (P < 0.02). This pattern was observed at every level of insulin sensitivity (P < 0.02). These data translated to an unfavorable (leftward) shift in the insulin feedback system for increasing 2-h glucose level (P < 0.005). Increased 2-h plasma glucose within the range of normal glucose tolerance in obese youth is associated with a specific impairment of beta-cell responsiveness distinct from the deterioration of insulin sensitivity. PMID- 15522933 TI - Gene expression analysis reveals evidence for increased expression of cell cycle associated genes and Gq-protein-protein kinase C signaling in cold thyroid nodules. AB - In contrast to the molecular etiology of autonomously functioning thyroid nodules, the molecular cause of cold thyroid nodules (CTNs), their benign, functional inactive counterparts, are so far largely unknown. Because of the partially dedifferentiated phenotype of CTNs, alterations in signaling cascades that favor proliferation, but not differentiation, are likely candidates for tumor induction and progression. The importance of RAS mutations for the development of benign nodules with follicular histology is still in question. However, differentially expressed genes in the context of their signaling cascades could define aberrant signaling in CTNs. Therefore, we investigated gene expression in 22 CTNs and their normal surrounding tissue using Affymetrix GeneChips. Most prominently, data analysis revealed an increased expression of cell cycle-associated genes and a special relevance of protein kinase C signaling, whereas no evidence of RAS-MAPK signaling in CTNs was found. Moreover, we determined 31 differentially regulated genes in CTNs, including several histone mRNAs. Taken together, these results explain recent findings showing an increased proliferation in CTNs and draw attention to protein kinase C signaling, but away from RAS-MAPK signaling, as being involved in the etiology of CTNs. PMID- 15522934 TI - Prostaglandin dehydrogenase and prostaglandin levels in periovulatory follicles: implications for control of primate ovulation by prostaglandin E2. AB - Prostaglandin (PG) E2 produced by the periovulatory follicle in response to the midcycle LH surge is essential for successful ovulation in primates. Granulosa cells express the PG synthesis enzyme cyclooxygenase-2 in response to the LH surge, but elevated cyclooxygenase-2 mRNA levels precede rising follicular fluid PGE2 levels by 24 h. Therefore, PG metabolism may play a significant role in regulating follicular concentrations of PGE2 during the periovulatory interval. To test this hypothesis, granulosa cells, follicular fluid, and whole ovaries were obtained from adult monkeys receiving exogenous gonadotropins to stimulate development of multiple, large follicles at times spanning the 40-h periovulatory interval. Ovarian expression of the NAD+-dependent 15-hydroxy PG dehydrogenase (PGDH) was assessed by RT-PCR, Western blotting, and immunohistochemistry. PGDH mRNA levels were low in granulosa cells obtained 0 h after hCG, rose 10-fold 12 h after hCG, and were not different from 0 h by 24-36 h after hCG administration. Granulosa cell PGDH protein was present 0-12 h after hCG but was low/nondetectable 36 h after hCG administration. Follicular fluid PGE2 levels were low at 0-12 h, slightly higher at 24 h, and then rose 10-fold to peak at 36 h hCG. Levels of biologically inactive PGE2 metabolites in follicular fluid were also low at 0 h but elevated at 12-24 h after hCG, times at which PGE2 levels remain low. Therefore, PGDH is present in the primate periovulatory follicle in a pattern consistent with modulation of follicular PGE2 levels during the periovulatory interval, supporting the hypothesis that gonadotropin-regulated PGDH plays a role in the control and timing of ovulation in primates. PMID- 15522935 TI - Postprandial plasma ghrelin is suppressed proportional to meal calorie content in normal-weight but not obese subjects. AB - Circulating levels of the gastric hormone ghrelin rise before and decrease after a meal. In normal-weight subjects, postprandial suppression of ghrelin is proportional to calories consumed. Obese individuals have lower fasting ghrelin levels; however, it is unclear whether the obese show normal postprandial suppression. This study aimed to compare postprandial ghrelin responses in normal weight and obese subjects, using mixed macronutrient meals with varied fat and calorie content. Postprandial ghrelin response was measured in normal-weight insulin-sensitive subjects and obese insulin-resistant subjects, after six test meals with different fat and calorie content (250-3000 kcal). Increasing the calorie content of meals in normal-weight subjects progressively lowered nadir levels of ghrelin. The obese had lower fasting ghrelin levels, and the reduction after the consumption of all test meals was less than the normal-weight subjects. The lowest postprandial levels in the obese were no different to the nadir in normal-weight volunteers after 1000-, 2000-, and 3000-kcal meals. Thus, circulating ghrelin levels decreased in normal-weight subjects after mixed meals. Obese subjects demonstrated a much reduced ghrelin postprandial suppression. This reduced suppression may influence satiety, thus reinforcing obesity. PMID- 15522936 TI - Requirement for proprotein convertase 5/6 during decidualization of human endometrial stromal cells in vitro. AB - Decidualization of endometrial stromal cells (ESCs) is critical for embryo implantation and maintenance of pregnancy. Proprotein convertase (PC) 5/6 is suggested to play an important role in the processes of stromal cell decidualization and embryo implantation in the mouse. PC5/6 is a member of the PC family responsible for processing precursor proteins to their active forms by selective proteolysis. In this study, we investigated the regulation of PC5/6 mRNA and protein expression in human ESCs during decidualization in vitro. Real time PCR analyses revealed a significant increase in PC5/6 mRNA levels in ESCs treated with 17 beta-estradiol (E(2)) plus medroxy-progesterone acetate during decidualization. On the other hand, E(2) alone did not increase PC5/6 mRNA expression. Intense PC5/6 immunoreactivity was observed in the cytoplasm of E(2) plus medroxy-progesterone acetate-treated ESCs (decidualized ESCs) compared with E(2)-treated ESCs on d 12 of culture (nondecidualized ESCs). This PC5/6 immunoreactivity was abolished by cotreatment with ZK 98299, a progesterone receptor antagonist. Western blotting revealed PC5/6 as approximately 120-kDa bands (pro- and mature forms) and a 65-kDa band (C-terminally truncated form) in decidualized ESCs. Using an antisense morpholino approach, prolactin production, a typical marker for decidualization, was significantly attenuated in decidualized ESCs after treatment with PC5/6 morpholino antisense oligonucleotides in comparison with controls. These results suggest that PC5/6 plays a key role for decidualization in human endometrium. PMID- 15522937 TI - Genetic variation at the locus encompassing 11-beta hydroxylase and aldosterone synthase accounts for heritability in cortisol precursor (11-deoxycortisol) urinary metabolite excretion. AB - Genetic variation in the gene encoding aldosterone synthase (CYP11B2) has previously been shown to be associated with hypertension and left ventricular hypertrophy. The intermediate phenotype most consistently associated with variation at this locus is that of elevated plasma 11-deoxycortisol (S). However, in normal subjects, aldosterone synthase does not metabolize S, which is converted to cortisol (F) by the enzyme 11 beta hydroxylase, encoded by the gene CYP11B1, which lies adjacent to CYP11B2 on chromosome 8. It is possible that the quantitative trait locus for the phenotype is within CYP11B1 and that linkage disequilibrium across the extended locus could account for these observations. However, variation across the whole CYP11B1/B2 locus had not been extensively characterized with respect to these phenotypes. We genotyped six polymorphisms in the CYP11B2 gene and three polymorphisms in the CYP11B1 gene in 248 Caucasian nuclear families comprising 1428 individuals. We measured plasma levels of S and F in 460 individuals from 86 families and urinary excretion rates of tetrahydrodeoxycortisol (THS) and tetrahydrodeoxycorticosterone in 573 individuals from 105 families. We examined heritability of the phenotypes and their association with genotypes and haplotypes at this locus. All steroid phenotypes except urinary tetrahydrodeoxycorticosterone were highly heritable (P < 0.00001). There was strong linkage disequilibrium across the CYP11B1/B2 locus. There was modest evidence for association between polymorphisms of CYP11B2 and plasma levels of S (P = 0.02 for T4986C polymorphism) and the plasma S to F ratio, reflecting the activity of 11-beta hydroxylase (P = 0.01 for T4986C polymorphism). There was strong evidence for association between polymorphisms of both CYP11B1 and CYP11B2 and urinary THS, which was strongest for the CYP11B1 exon 1 polymorphism (P = 0.00002). Addition of other marker data to CYP11B1 exon 1 did not improve the fit of a log-linear model. Genotype at CYP11B1 explained approximately 5% of the variance in urinary THS excretion in the population. Thus, it is likely that linkage disequilibrium between causative CYP11B1 variants and CYP11B2 polymorphisms account for the previous observations. Further fine mapping studies across the CYP11B1 locus are required to localize the causative variant(s) for the biochemical phenotype; this may also identify susceptibility alleles for hypertension and left ventricular hypertrophy. PMID- 15522939 TI - Giant insulinoma: case report and review of the literature. AB - An insulinoma is a rare pancreatic endocrine tumor that is typically sporadic, solitary, and less than 2 cm in diameter. Fewer than 5% of insulinomas are larger than 3 cm. Ninety percent or more of all insulinomas are benign. Larger tumors are more likely to be malignant. We report a case of a giant pedunculated insulinoma, measuring 9 cm in diameter and weighing 100 g, with amyloid deposits accounting for 70% of the tumor volume. At the time of operation, no local invasion or metastatic disease was identified. On pathological evaluation, the tumor was classified as an insulinoma of uncertain biological behavior. In addition to describing the clinical presentation and operative findings, criteria for determining malignancy are outlined, a detailed pathological description is presented, and the 2000 World Health Organization Classification for Pancreatic Endocrine Neoplasms is reviewed. PMID- 15522938 TI - Cinacalcet hydrochloride maintains long-term normocalcemia in patients with primary hyperparathyroidism. AB - Calcimimetics increase the sensitivity of parathyroid calcium-sensing receptors to extracellular calcium, thereby reducing PTH secretion. This multicenter, randomized, double-blind, placebo-controlled study assessed the ability of the oral calcimimetic cinacalcet HCl to achieve long-term reductions in serum calcium and PTH concentrations in patients with primary hyperparathyroidism (HPT). Patients (n = 78) were randomized to cinacalcet or placebo. Cinacalcet was titrated from 30-50 mg twice daily during a 12-wk dose-titration phase. Efficacy was assessed during 12-wk maintenance and 28-wk follow-up phases. The primary endpoint was the achievement of normocalcemia [serum calcium 30 (obese men (OM), 41 (30-72) kg/m(2) (n = 15) and women (OW), 43 (30-68) kg/m(2) (n = 34)]. Fat mass and percentage fat were measured by a bioelectrical impedance analyzer. An IGF-I generation test, which involved a sc injection of 21 IU (7 mg) GH, was performed. At baseline serum samples were assayed for GHBP; serum IGF-I and IGFBP3 levels were measured both at baseline and 24 h after GH administration. There was a higher increment IGF-I in obese men and women, compared with the equivalent normal-weight subjects [NM vs. OM: 245 (33-342) vs. 291 (192-427) ng/ml (P < 0.05); NW vs. OW: 220 (103-435) vs. 315 (144-450) ng/ml (P < 0.0005)]. Increment IGF-I was negatively correlated with baseline IGF-I (F = 12.1) and positively correlated with GHBP (F = 18.2) (R(2) = 0.29). GHBP levels were significantly higher in OM and OW (pre- and postmenopausal) than in the equivalent normal-weight groups [NM vs. OM: 2175 (995 4190) vs. 3030 (1540-5470) pmol/liter (P < 0.05); NW vs. OW: 2131 (1010-5040) vs. 3585 (1540-5740) pmol/liter (P < 0.0005)]. GHBP levels correlated highly with BMI, percentage fat, and fat mass (R > 0.6, P < 0.0001). Baseline IGF-I was not affected by body composition. In conclusion, in obese compared with normal-weight healthy subjects, there is a larger increment IGF-I to a single bolus of GH in men, and irrespective of menopausal status, women. Increment IGF-I is associated positively with GHBP level, which in turn is associated with markers of increasing obesity in men and women. GH responsiveness is increased in obesity. PMID- 15522944 TI - Androgen insensitivity syndrome: somatic mosaicism of the androgen receptor in seven families and consequences for sex assignment and genetic counseling. AB - Androgen insensitivity syndrome (AIS) is caused by numerous mutations of the androgen receptor (AR) gene. The phenotype may range from partial AIS (PAIS) with ambiguous genitalia to complete AIS (CAIS) with female genitalia. In 70% of the cases, AR mutations are transmitted in an X-linked recessive manner through the carrier mothers, but in 30%, the mutations arise de novo. When de novo mutations occur after the zygotic stage, they result in somatic mosaicisms, which are an important consideration for both virilization in later life-because both mutant and wild-type receptors are expressed-and genetic counseling. We report here six patients with AIS due to somatic mutations of the AR and one mother with somatic mosaicism who transmitted the mutation twice. Of the four patients with PAIS, three presented spontaneous or induced virilization at birth or puberty. These cases underline the crucial role of the remnant wild-type AR for virilization because the same mutations, when they are inherited, lead to CAIS. We also report two novel mutations of the AR, with somatic mosaicism, detected in patients with CAIS. Thus, the remnant wild-type receptor does not always lead to virilization. In one of these patients, a high ratio of wild-type to mutant AR expression was found in the gonads and genital skin fibroblasts. Although no prenatal virilization occurred, the possibility of virilization at puberty could not be excluded, and early gonadectomy was performed. A seventh patient had a CAIS with a novel germline AR mutation. The mutation was inherited from the mother, in whom mosaicism was detected in blood and who transmitted the mutation to a second, XX, offspring. The detection of somatic AR mutations is particularly important for the clinical management and genetic counseling of patients with AIS. Before definite sex assignment, a testosterone treatment trial should be performed in all patients with PAIS, but it becomes crucial when an AR mosaicism has been detected. In patients with CAIS or severe PAIS raised as female, there is no consensus about when (early childhood or puberty) gonadectomy should be performed. When somatic AR mutations are detected, however, gonadectomy should be performed earlier because of the risk of virilization during puberty. When a germline de novo mutation is identified in the index case, the risk of transmission to a second child due to a possible germ cell mosaicism in the mother cannot be excluded. However, given the high number of AR de novo mutations and the rarity of such reports, this risk appears to be very low. PMID- 15522945 TI - Expression of leptin receptors and potential effects of leptin on the cell growth and activation of mitogen-activated protein kinases in ovarian cancer cells. AB - Leptin, a secreted protein of the ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic breast cancer cells. However, a potential role of leptin as an endocrine regulator is unknown in ovarian cancer. In the present study, we investigated the expression of leptin receptors in immortalized ovarian surface epithelium (IOSE) and ovarian cancer cell lines, and potential effect of leptin on the cell growth and activation of mitogen-activated protein kinases (MAPKs) in the BG-1 ovarian cancer cell line. Both short and long isoforms of leptin receptors are expressed in IOSE-80PC (a post-crisis line), BG-1, OVCAR-3, and SKOV-3 cells. In addition, treatment with leptin resulted in the growth stimulation of BG-1 cells, an activation of ERK1/2 and inhibition of constitutive phosphorylation of p38 MAPK. These results suggest that further studies are necessary to validate whether leptin may be a potential regulator for ovarian cancer. PMID- 15522946 TI - National legislation on school smoking restrictions in eight European countries. AB - OBJECTIVE: To review and compare national legislation addressing smoking at school in eight European countries during the late 1990s. DESIGN: The data are from the EC-funded Control of Adolescent Smoking (CAS) study in eight European countries. The information on national legislation was gathered during the period 1998-1999, through a review of scientific and official documents and interviews with key informants in each country. RESULTS: Four of the participating countries (Austria, French-speaking Belgium, Finland and Norway) had legislation specifically restricting smoking at school, while the remaining countries (Denmark, North Rhein Westphalia region of Germany, Scotland and Wales in the UK) did not have such legislation in place. In those countries with legislation in place, smoking among students aged 15 years and younger was not permitted. The position with regard to teacher smoking varied considerably among countries, but with the exception of Finland, there was no legislation regulating outdoor smoking by teachers during school hours. CONCLUSIONS: The findings suggest that there were inconsistencies within countries in terms of legislation that had been developed and the enforcement, compliance and monitoring practices that were in place. Further work is required to develop and resource such mechanisms, although it may be possible to build on existing practice, such as increasing the involvement of school health services, school inspectorate services or working through health promoting school networks already established in many countries. PMID- 15522948 TI - What's the place of schools in promoting health? Are we too optimistic? PMID- 15522947 TI - Acceptability and feasibility of an interactive computer-tailored fat intake intervention in Belgium. AB - In order to reduce the risk of chronic diseases health authorities recommend restricting fat intake to 30% of the total energy uptake. However, fat intake in Belgium is much higher warranting interventions aimed at reducing fat intake. Tailored interventions have shown to be promising; however, studies on effectiveness of interactive computer-tailored systems are needed. We investigated the acceptability and feasibility of a recently developed interactive computer-tailored fat reduction intervention. Differences in the reported acceptability and feasibility according to demographic and stages of change were explored. Participants (n = 220) completed a computerized questionnaire, and received a personal fat intake advice, which was almost immediately displayed on screen. They also completed an evaluation questionnaire, during and after they ran the tailored program, with questions on the quality, user-friendliness and applicability of the program. Participants rated the program positively on all aspects. No significant differences in acceptability and feasibility scores were found according to sex, education levels and computer literacy. Although several significant differences were found between age groups and stages of change (oldest group, contemplators and preparators were more positive about the program), the importance of these differences is probably not great, since acceptability and feasibility scores for the different age groups and stages of change were always very high. These results suggest that the computer-tailored intervention is an acceptable and feasible tool for reducing fat intake in a general population in Belgium. PMID- 15522949 TI - ACE gene titration in mice uncovers a new mechanism for ACE on the control of body weight. AB - Mice harboring 1, 2, or 3 copies of the angiotensin-converting enzyme (ACE) gene were used to evaluate the quantitative role of the ACE locus on obesity. Three copy mice fed with a high-fat diet had lower body weight and peri-epididymal adipose tissue than did 1- and 2-copy mice (P < 0.05). On regular diet, 3-copy mice had to eat more to maintain the same body weight; on a high-fat diet, they ate the same but weighed less than 1- and 2-copy mice (P < 0.05), indicating a higher metabolic rate in 3-copy mice that was not affected by ANG II AT(1) blocker treatment. A catalytically inactive form of thimet oligopeptidase (EC 3.4.24.15; EP24.15) was used to isolate ACE substrates from adipose tissue. Liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI MS/MS) identified 162 peptide peaks; 16 peptides were present in both groups (1- and 3-copy mice fed with a high-fat diet), whereas 58 of the 72 unique peptides were found only in the 3-copy mice. Peptide size distribution was shifted to lower molecular weight in 3-copy mice. Two of the identified peptides, LVVYPWTQRY and VVYPWTQRY, which are ACE substrates, inhibited in vitro protein kinase C phosphorylation in a concentration-dependent manner. In addition, neurolysin (EC 3.4.24.16; EP24.16) activity was lower in fat tissue from 3- vs. 1-copy mice (P < 0.05). Taken together, these results provide evidence that ACE is associated with body weight and peri-epididymal fat accumulation. This response may involve the generation of oligopeptides that inhibit the activity of EP24.16 and other oligopeptidases within the adipose tissue. PMID- 15522950 TI - Identification of new targets of Drosophila pre-mRNA adenosine deaminase. AB - Adenosine deaminase acting on RNA (ADAR) in Drosophila and mammals has recently become the target of numerous investigations. It is now clear that this protein has a number of functions in the nervous system. Indeed, the mutation of ADAR in Drosophila (dADAR) results in many pathological and physiological changes, such as sensitivity to hypoxia and neuronal degeneration. To understand the full scope of dADAR function, it is crucial to identify new dADAR targets. A polyclonal antibody against inosine was developed and used to enrich inosine-containing mRNAs. The efficiency of immunoaffinity purification was confirmed for the Q/R editing site of GluR-B pre-mRNA that has been edited by ADAR2 to generate inosines at the editing site. This approach was applied to enrich inosine containing mRNAs from total mRNAs of wild-type and dADAR mutant flies, respectively. The enriched mRNA portion was then amplified and hybridized with Drosophila cDNA arrays. With this method, over 500 mRNAs were identified as potential dADAR targets by showing a higher amount in the enriched mRNA portion from wild-type flies than from dADAR mutant flies. The occurrence of A-to-G conversion in these mRNAs was further analyzed by comparing over 7,000 Drosophila cDNAs sequences with their genomic sequences. A final list of 62 candidates was generated from the overlap of the two approaches. Twelve genes from the final list were further examined by sequencing the RT-PCR products of these genes from wild-type and dADAR mutant flies. Seven of the 12 genes were proven to have A-to G changes in the wild-type but not in mutant flies. We conclude that the combination of immunoaffinity enrichment of inosine-containing mRNA, DNA microarrays, and sequence comparison could facilitate the discovery of new dADAR substrates, which in turn allows us to better understand the targets of dADAR and the biological function of A-to-I RNA editing in flies. PMID- 15522951 TI - Prospective isolation and global gene expression analysis of the erythrocyte colony-forming unit (CFU-E). AB - The erythrocyte colony-forming unit (CFU-E) is a rare bone marrow (BM) progenitor that generates erythrocyte colonies in 48 hours. The existence of CFU-Es is based on these colonies, but CFU-Es have not been purified prospectively by phenotype. We have separated the "nonstem," "nonlymphoid" compartment (lineage marker [lin] c-Kit+Sca-1-IL-7Ralpha-) into interleukin 3 receptor alpha negative (IL-3Ralpha-) and IL-3Ralpha+ subsets. Within IL-3Ralpha- but not IL-3Ralpha+ cells we have identified TER119-CD41-CD71+ erythrocyte-committed progenitors (EPs). EPs generate CFU-E colonies at about 70% efficiency and generate reticulocytes in vivo. Depletion of EPs from BM strongly reduces CFU-E frequencies. EPs lack potential for erythrocyte burst-forming unit, megakaryocyte, granulocyte (G), and monocyte (M) colonies, and for spleen colony-forming units. Chronically suppressed erythropoiesis in interferon consensus sequence-binding protein (ICSBP)-deficient BM is associated with reduced frequencies of both the EP population and CFU-E colonies. During phenylhydrazine-induced acute anemia, numbers of both the EP population and CFU-E colonies increase. Collectively, EPs (lin-c-Kit+Sca-1-IL-7Ralpha-IL-3Ralpha-CD41-CD71+) account for most, if not all, CFU-E activity in BM. As a first molecular characterization, we have compared global gene expression in EPs and nonerythroid GM progenitors. These analyses define an erythroid progenitor-specific gene expression pattern. The prospective isolation of EPs is an important step to analyze physiologic and pathologic erythropoiesis. PMID- 15522952 TI - The earliest subpopulation of mouse thymocytes contains potent T, significant macrophage, and natural killer cell but no B-lymphocyte potential. AB - The B-lymphocyte potential of progenitor thymocytes and whether the thymus is colonized by common lymphocyte progenitor cells have been subjects of considerable debate. Herein, we have used limiting dilution analysis to determine the lineage potential of phenotypically defined subpopulations of CD4-CD8- double negative thymocytes. Culture systems used showed single-hit kinetics and had a high plating efficiency for B-, myeloid, and natural killer cell development. The T-cell potential of sorted cells was confirmed by transferring cells to fetal thymus organ cultures. Our results indicate that the earliest population of CD117+ double-negative cells, although containing potent T-cell developmental potential and significant myeloid and natural killer potential, does not have any residual B-cell potential. Gene transcription analysis also indicated that these double-negative cells contained abundant T and myeloid, but not B cell-specific transcripts. The implications of these results within the context of current models of thymocyte development are discussed. PMID- 15522953 TI - Hematopoietic stem cells from NOD mice exhibit autonomous behavior and a competitive advantage in allogeneic recipients. AB - Type 1 diabetes is a systemic autoimmune disease that can be cured by transplantation of hematopoietic stem cells (HSCs) from disease-resistant donors. Nonobese diabetic (NOD) mice have a number of features that distinguish them as bone marrow transplant recipients that must be understood prior to the clinical application of chimerism to induce tolerance. In the present studies, we characterized NOD HSCs, comparing their engraftment characteristics to HSCs from disease-resistant strains. Strikingly, NOD HSCs are significantly enhanced in engraftment potential compared with HSCs from disease-resistant donors. Unlike HSCs from disease-resistant strains, they do not require graft-facilitating cells to engraft in allogeneic recipients. Additionally, they exhibit a competitive advantage when coadministered with increasing numbers of syngeneic HSCs, produce significantly more spleen colony-forming units (CFU-Ss) in vivo in allogeneic recipients, and more granulocyte macrophage-colony-forming units (CFU-GMs) in vitro compared with HSCs from disease-resistant controls. NOD HSCs also exhibit significantly enhanced chemotaxis to a stromal cell-derived factor 1 (SDF-1) gradient and adhere significantly better on primary stroma. This enhanced engraftment potential maps to the insulin-dependent diabetes locus 9 (Idd9) locus, and as such the tumor necrosis factor (TNF) receptor family as well as ski/sno genes may be involved in the mechanism underlying the autonomy of NOD HSCs. These findings may have important implications to understand the evolution of autoimmune disease and impact on potential strategies for cure. PMID- 15522954 TI - Overexpression of mitochondrial ferritin causes cytosolic iron depletion and changes cellular iron homeostasis. AB - Cytosolic ferritin sequesters and stores iron and, consequently, protects cells against iron-mediated free radical damage. However, the function of the newly discovered mitochondrial ferritin (MtFt) is unknown. To examine the role of MtFt in cellular iron metabolism, we established a cell line that stably overexpresses mouse MtFt under the control of a tetracycline-responsive promoter. The overexpression of MtFt caused a dose-dependent iron deficiency in the cytosol that was revealed by increased RNA-binding activity of iron regulatory proteins (IRPs) along with an increase in transferrin receptor levels and decrease in cytosolic ferritin. Consequently, the induction of MtFt resulted in a dramatic increase in cellular iron uptake from transferrin, most of which was incorporated into MtFt. The induction of MtFt caused a shift of iron from cytosolic ferritin to MtFt. In addition, iron inserted into MtFt was less available for chelation than that in cytosolic ferritin and the expression of MtFt was associated with decreased mitochondrial and cytosolic aconitase activities, the latter being consistent with the increase in IRP-binding activity. In conclusion, our results indicate that overexpression of MtFt causes a dramatic change in intracellular iron homeostasis and that shunting iron to MtFt likely limits its availability for active iron proteins. PMID- 15522955 TI - Neuropilin-1 regulates attachment in human endothelial cells independently of vascular endothelial growth factor receptor-2. AB - Neuropilin-1 (NRP-1) is a type 1 membrane protein that binds the axon guidance factors belonging to the class-3 semaforin family. In endothelial cells, NRP-1 serves as a co-receptor for vascular endothelial growth factor (VEGF) and regulates VEGF receptor 2 (VEGFR-2)-dependent angiogenesis. Although gene targeting studies documenting embryonic lethality in NRP-1 null mice have demonstrated a critical role for NRP-1 in vascular development, the activities of NRP-1 in mature endothelial cells have been incompletely defined. Using RNA interference-mediated silencing of NRP-1 or VEGFR-2 in primary human endothelial cells, we confirm that NRP-1 modulates VEGFR-2 signaling-dependent mitogenic functions of VEGF. Importantly, we now show that NRP-1 regulates endothelial cell adhesion to extracellular matrix proteins independently of VEGFR-2. Based on its dual role as an enhancer of VEGF activity and a mediator of endothelial cell adhesiveness described here, NRP-1 emerges as a promising molecular target for the development of antiangiogenic drugs. PMID- 15522956 TI - A common founder mutation in FANCA underlies the world's highest prevalence of Fanconi anemia in Gypsy families from Spain. AB - Fanconi anemia (FA) is a genetic disease characterized by bone marrow failure and cancer predisposition. Here we have identified Spanish Gypsies as the ethnic group with the world's highest prevalence of FA (carrier frequency of 1/64-1/70). DNA sequencing of the FANCA gene in 8 unrelated Spanish Gypsy FA families after retroviral subtyping revealed a homozygous FANCA mutation (295C>T) leading to FANCA truncation and FA pathway disruption. This mutation appeared specific for Spanish Gypsies as it is not found in other Gypsy patients with FA from Hungary, Germany, Slovakia, and Ireland. Haplotype analysis showed that Spanish Gypsy patients all share the same haplotype. Our data thus suggest that the high incidence of FA among Spanish Gypsies is due to an ancestral founder mutation in FANCA that originated in Spain less than 600 years ago. The high carrier frequency makes the Spanish Gypsies a population model to study FA heterozygote mutations in cancer. PMID- 15522957 TI - Deregulated expression of the Myc cellular oncogene drives development of mouse "Burkitt-like" lymphomas from naive B cells. AB - Chromosomal translocations juxtaposing immunoglobulin (Ig) and MYC genes are the hallmarks of human Burkitt lymphoma (BL), with deregulated MYC expression being a critical factor in pathogenesis. By inserting an intact mouse Myc gene into the mouse genome, proximal to the Ig enhancer Emu, the effect of a precise mimic of the major t(8;14) translocation of human endemic BL (eBL) could be investigated. Knock-in mice developed IgM-positive B-cell tumors, with most being typical of eBL by histology and immunophenotype, including expression of the germinal center (GC)-associated protein, BCL6. Unlike eBL, however, analysis of Ig V(H) sequences revealed no significant level of somatic mutation. Thus, constitutive expression of Myc in the knock-in mice is apparently able to induce "Burkitt-like" lymphomas before antigen stimulation and formation of a GC. In contrast, human eBL development occurs in a GC or post-GC site with a likely contribution to pathogenesis from Epstein-Barr virus (EBV) and other epigenetic factors. PMID- 15522958 TI - Stimulation with 4-1BB (CD137) inhibits chronic graft-versus-host disease by inducing activation-induced cell death of donor CD4+ T cells. AB - 4-1BB, a member of the tumor necrosis factor (TNF) receptor superfamily, is a costimulator for activated T cells. Previous studies have established that treatment with agonistic anti-4-BB monoclonal antibody (3H3) is effective in reversing the progression of spontaneous systemic lupus erythematosus. Its therapeutic effect is mediated by suppression of autoantibody production. In this report, we show that a single injection of 3H3 blocks chronic graft-versus-host disease (cGVHD) in the parent-into-F1 model. In particular, donor CD4+ T cells are rapidly eliminated from host spleens by activation-induced cell death after 4 1BB triggering. Since donor CD4+ T cells are required for the development of cGVHD, and 3H3-mediated inhibition of autoantibody production occurs without donor CD8+ T cells, 3H3 blocks cGVHD by preventing alloreactive donor CD4+ T cells from activating host B cells. Importantly, 3H3 treatment can reverse the progression of advanced cGVHD. Our findings indicate that agonistic anti-4-1BB monoclonal antibody has potential as an immunotherapeutic agent for preventing and treating cGVHD. PMID- 15522959 TI - HOXB6 overexpression in murine bone marrow immortalizes a myelomonocytic precursor in vitro and causes hematopoietic stem cell expansion and acute myeloid leukemia in vivo. AB - The HOX family of homeobox genes plays an important role in normal and malignant hematopoiesis. Dysregulated HOX gene expression profoundly effects the proliferation and differentiation of hematopoietic stem cells (HSCs) and committed progenitors, and aberrant activation of HOX genes is a common event in human myeloid leukemia. HOXB6 is frequently overexpressed in human acute myeloid leukemia (AML). To gain further insight into the role of HOXB6 in hematopoiesis, we overexpressed HOXB6 in murine bone marrow using retrovirus-mediated gene transfer. We also explored structure-function relationships using mutant HOXB6 proteins unable to bind to DNA or a key HOX-binding partner, pre-B-cell leukemia transcription factor-1 (PBX1). Additionally, we investigated the potential cooperative interaction with myeloid ecotropic viral integration site 1 homolog (MEIS1). In vivo, HOXB6 expanded HSCs and myeloid precursors while inhibiting erythropoiesis and lymphopoiesis. Overexpression of HOXB6 resulted in AML with a median latency of 223 days. Coexpression of MEIS1 dramatically shortened the onset of AML. Cytogenetic analysis of a subset of HOXB6-induced AMLs revealed recurrent deletions of chromosome bands 2D-E4, a region frequently deleted in HOXA9-induced AMLs. In vitro, HOXB6 immortalized a factor-dependent myelomonocytic precursor capable of granulocytic and monocytic differentiation. These biologic effects of HOXB6 were largely dependent on DNA binding but independent of direct interaction with PBX1. PMID- 15522960 TI - High throughput mRNA profiling highlights associations between myocardial infarction and aberrant expression of inflammatory molecules in blood cells. AB - Studies on the role of inflammation in cardiovascular disease focus on surrogate markers like plasma levels of C-reactive protein or interleukins that are affected by several factors. In this study we employ an approach in which the inflammatory mRNA profile of leucocytes is measured directly in a multigene system. We investigated the mRNA profile for 35 inflammatory markers in blood samples in a case-control study including 524 men with a history of myocardial infarction and 628 control subjects. Compared with controls, patients showed mRNA profiles with increased levels of most inflammatory mRNAs. The 2 most prominent mRNA risk indicators encoded the secreted protein macrophage migration inhibitory factor (crude odds ratio [OR], 3.4 for the highest quartile versus the lowest quartile (95% confidence interval [CI95], 2.3-4.9), and the intracellular regulator proteinase inhibitor 9 (OR, 2.5 for the highest versus the lowest quartile (CI95, 1.8-3.5), both showing an increase in odds ratio with increasing quartiles. Leucocytes in the blood of patients with myocardial infarction are more active in transcription of inflammatory genes, as evidenced by mRNA profiling. These data support the hypothesis that an inflammatory response involving leucocytes plays a role in the pathogenesis of myocardial infarction. PMID- 15522961 TI - Distinct roles for donor- and host-derived antigen-presenting cells and costimulatory molecules in murine chronic graft-versus-host disease: requirements depend on target organ. AB - The application of allogeneic stem cell transplantation (alloSCT) is limited by graft-versus-host disease (GVHD). GVHD can be divided into acute and chronic forms that likely have different requirements for initiation and pathogenesis mechanisms. In prior studies we demonstrated that residual host antigen presenting cells (APCs) were required to initiate acute GVHD (aGVHD) mediated by CD8 T cells. In contrast, here we demonstrate that either donor or host APCs can initiate CD4-mediated GVHD in a model that has features of chronic GVHD (cGVHD). Both donor and host APCs must provide CD80/86-dependent costimulation to elicit maximal cGVHD, and there is no GVHD when both donor and host lack CD80/86. Finally, we were surprised to find that, although either donor or host APCs are sufficient to stimulate skin cGVHD, donor APCs play a dominant role in intestinal cGVHD. Both CD40 and CD80/86 are critical for donor APC function in intestinal cGVHD, but only CD80/86 is required for skin cGVHD. Thus, there are target-tissue specific differences in APC requirements. These results identify differences in APC requirements between CD8-mediated aGVHD and CD4-mediated cGVHD. They further highlight donor APCs as additional targets for GVHD therapy. PMID- 15522962 TI - C1 inhibitor prevents Gram-negative bacterial lipopolysaccharide-induced vascular permeability. AB - Gram-negative bacterial endotoxemia may lead to the pathological increase of vascular permeability with systemic vascular collapse, a vascular leak syndrome, multiple organ failure (MOF), and/or shock. Previous studies demonstrated that C1 inhibitor (C1INH) protects mice from lipopolysaccharide (LPS)-induced lethal septic shock via a direct interaction with LPS. Here, we report that C1INH blocked the LPS-induced increase in transendothelial flux through an endothelial monolayer. In addition, LPS-mediated detachment of cultured endothelial cells was prevented with C1INH. C1INH also inhibited LPS-induced endothelial cell apoptosis as demonstrated by suppression of DNA fragmentation and annexin V expression. As illustrated by laser scanning confocal microscopy, C1INH completely blocked the binding of fluorescein isothiocyanate (FITC)-LPS to human umbilical vein endothelial cells (HUVECs). C1INH protected from localized LPS-induced increased plasma leakage in C57BL/6J mice and in C1INH-deficient mice. Local vascular permeability in response to LPS was increased to a greater extent in C1INH deficient mice compared with wild-type littermate controls and was reversed by treatment with C1INH. Systemic administration of LPS to mice resulted in increased vascular permeability, which was reduced by C1INH. Therefore, these studies demonstrate that C1INH, in addition to its role in suppression of LPS mediated macrophage activation, may play an important role in the prevention of LPS-mediated increased vascular permeability, endothelial cell injury, and multiple organ failure. PMID- 15522963 TI - An erythroid differentiation-specific splicing switch in protein 4.1R mediated by the interaction of SF2/ASF with an exonic splicing enhancer. AB - Protein 4.1R is a vital component of the red blood cell membrane cytoskeleton. Promotion of cytoskeletal junctional complex stability requires an erythroid differentiation stage-specific splicing switch promoting inclusion of exon 16 within the spectrin/actin binding domain. We showed earlier that an intricate combination of positive and negative RNA elements controls exon 16 splicing. In this report, we further identified 3 putative exonic splicing enhancers within exon 16 and investigated the function of the sequence CAGACAT in the regulation of exon 16 splicing. Mutation of these sequences leads to increased exclusion of exon 16 in both in vivo and in vitro splicing assays, indicating that CAGACAT is a functional exonic splicing enhancer. UV cross-linking further detects an approximately 33-kDa protein that specifically binds to the CAGACAT-containing transcript. An anti-SF2/ASF antibody specifically immunoprecipitates the approximately 33-kDa protein. Furthermore, SF2/ASF stimulates exon 16 inclusion in both in vitro complementation assays and minigene-transfected mouse erythroleukemia cells (MELCs). Finally, SF2/ASF expression is up-regulated and correlates with exon 16 inclusion in differentiated MELCs. These results suggest that increased splicing factor 2/alternative splicing factor (SF2/ASF) expression in differentiated mouse erythroleukemia mediates a differentiation stage-specific exon 16 splicing switch through its interaction with the exonic splicing enhancer. PMID- 15522964 TI - Thrombin induces endocytosis of endoglin and type-II TGF-beta receptor and down regulation of TGF-beta signaling in endothelial cells. AB - Thrombin activates protease-activated receptor 1 (PAR1) on endothelial cells (ECs) and is critical for angiogenesis and vascular development. However, the mechanism underlying the proangiogenic effect of thrombin has not been elucidated yet. Here, we report the discovery of a novel functional link between thrombin PAR1 and transforming growth factor-beta (TGF-beta) signaling pathways. We showed that thrombin via PAR1 induced the internalization of endoglin and type-II TGF beta receptor (TbetaRII) but not type-I receptors in human ECs. This effect was mediated by protein kinase C-zeta (PKC-zeta) since specific inhibition of PKC zeta caused an aggregation of endoglin or TbetaRII on cell surface and blocked their internalization by thrombin. Furthermore, acute and long-term pretreatment of ECs with thrombin or PAR1 peptide agonist suppressed the TGF-beta-induced serine phosphorylation of Smad2, a critical mediator of TGF-beta signaling. Moreover, activation of PAR1 led to a profound and spread cytosolic clustering formation of Smad2/3 and markedly prevented Smad2/3 nuclear translocation evoked by TGF-beta1. Since TGF-beta plays a crucial role in the resolution phase of angiogenesis, the down-regulation of TGF-beta signaling by thrombin-PAR1 pathway may provide a new insight into the mechanism of the proangiogenic effect of thrombin. PMID- 15522965 TI - Tissue plasminogen activator promotes the effects of corticotropin-releasing factor on the amygdala and anxiety-like behavior. AB - Stress-induced plasticity in the brain requires a precisely orchestrated sequence of cellular events involving novel as well as well known mediators. We have previously demonstrated that tissue plasminogen activator (tPA) in the amygdala promotes stress-induced synaptic plasticity and anxiety-like behavior. Here, we show that tPA activity in the amygdala is up-regulated by a major stress neuromodulator, corticotropin-releasing factor (CRF), acting on CRF type-1 receptors. Compared with WT, tPA-deficient mice responded to CRF treatment with attenuated expression of c-fos (an indicator of neuronal activation) in the central and medial amygdala but had normal c-fos responses in paraventricular nuclei. They exhibited reduced anxiety-like behavior to CRF but had a sustained corticosterone response after CRF administration. This effect of tPA deficiency was not mediated by plasminogen, because plasminogen-deficient mice demonstrated normal behavioral and hormonal changes to CRF. These studies establish tPA as an important mediator of cellular, behavioral, and hormonal responses to CRF. PMID- 15522966 TI - Cell-density-dependent regulation of neural precursor cell function. AB - Stress-induced reductions of neural precursor cells from the subgranular zone of the hippocampal dentate gyrus have been linked to impaired neurogenesis and cognitive dysfunction. Given the importance of redox state in regulating multiple damage-responsive pathways in the CNS, we hypothesize that oxidative stress plays a major role in affecting neurogenesis and subsequent cognitive function after cell injury/depletion. Using an in vitro system, we showed that the level of reactive oxygen species (ROS), which depend critically on changes in cell density, were significantly higher in neural precursor cells when compared with other primary and transformed cell lines. ROS were significantly elevated ( approximately 4-fold) under low- (<1 x 10(4) cells per cm(2)) versus high-density (>1 x 10(5) cells per cm(2)) conditions. Higher ROS levels found at lower cell densities were associated with elevated proliferation and increased metabolic activity. These ROS were likely a result of altered mitochondrial function that ultimately compromised the growth rate of cells. At high cell densities, intracellular ROS and oxidative damage were reduced in concert with an increased expression of mitochondrial superoxide dismutase 2. Our finding that DNA-damage induced depletion of neural precursor cells in the subgranular zone of mice also led to increased ROS and altered proliferation validated our in vitro system. Increased ROS and proliferation associated with the reduction of precursor cell numbers both in vitro and in vivo could be reversed with the antioxidant alpha lipoic acid. These data showed that neural precursor cells were predisposed to microenvironmental cues that regulate redox-sensitive pathways to control cellular proliferation after CNS damage. PMID- 15522967 TI - Model for anaphase B: role of three mitotic motors in a switch from poleward flux to spindle elongation. AB - It has been proposed that the suppression of poleward flux within interpolar microtubule (ipMT) bundles of Drosophila embryonic spindles couples outward forces generated by a sliding filament mechanism to anaphase spindle elongation. Here, we (i) propose a molecular mechanism in which the bipolar kinesin KLP61F persistently slides dynamically unstable ipMTs outward, the MT depolymerase KLP10A acts at the poles to convert ipMT sliding to flux, and the chromokinesin KLP3A inhibits the depolymerase to suppress flux, thereby coupling ipMT sliding to spindle elongation; (ii) used KLP3A inhibitors to interfere with the coupling process, which revealed an inverse linear relation between the rates of flux and elongation, supporting the proposed mechanism and demonstrating that the suppression of flux controls both the rate and onset of spindle elongation; and (iii) developed a mathematical model using force balance and rate equations to describe how motors sliding the highly dynamic ipMTs apart can drive spindle elongation at a steady rate determined by the extent of suppression of flux. PMID- 15522968 TI - Two important polymers cross paths. PMID- 15522969 TI - Visualization of the movement of single histidine kinase molecules in live Caulobacter cells. AB - The bacterium Caulobacter crescentus divides asymmetrically as part of its normal life cycle. This asymmetry is regulated in part by the membrane-bound histidine kinase PleC, which localizes to one pole of the cell at specific times in the cell cycle. Here, we track single copies of PleC labeled with enhanced yellow fluorescent protein (EYFP) in the membrane of live Caulobacter cells over a time scale of seconds. In addition to the expected molecules immobilized at one cell pole, we observed molecules moving throughout the cell membrane. By tracking the positions of these molecules for several seconds, we determined a diffusion coefficient (D) of 12 +/- 2 x 10(-3) microm(2)/s for the mobile copies of PleC not bound at the cell pole. This D value is maintained across all cell cycle stages. We observe a reduced D at poles containing localized PleC-EYFP; otherwise D is independent of the position of the diffusing molecule within the bacterium. We did not detect any directional bias in the motion of the PleC-EYFP molecules, implying that the molecules are not being actively transported. PMID- 15522970 TI - Folding of human superoxide dismutase: disulfide reduction prevents dimerization and produces marginally stable monomers. AB - The molecular mechanism by which the homodimeric enzyme Cu/Zn superoxide dismutase (SOD) causes neural damage in amytrophic lateral sclerosis is yet poorly understood. A striking, as well as an unusual, feature of SOD is that it maintains intrasubunit disulfide bonds in the reducing environment of the cytosol. Here, we investigate the role of these disulfide bonds in folding and assembly of the SOD apo protein (apoSOD) homodimer through extensive protein engineering. The results show that apoSOD folds in a simple three-state process by means of two kinetic barriers: 2D<==>2M<==>M(2). The early predominant barrier represents folding of the monomers (M), and the late barrier the assembly of the dimer (M(2)). Unique for this mechanism is a dependence of protein concentration on the unfolding rate constant under physiological conditions, which disappears above 6 M Urea where the transition state for unfolding shifts to first-order dissociation of the dimer in accordance with Hammond-postulate behavior. Although reduction of the intrasubunit disulfide bond C57-C146 is not critical for folding of the apoSOD monomer, it has a pronounced effect on its stability and abolishes subsequent dimerization. Thus, impaired ability to form, or retain, the C57-C146 bond in vivo is predicted to increase the cellular load of marginally stable apoSOD monomers, which may have implications for the amytrophic lateral sclerosis neuropathology. PMID- 15522975 TI - Generalized exfoliative dermatitis induced by interferon alfa. PMID- 15522971 TI - A clock gene, period, plays a key role in long-term memory formation in Drosophila. AB - The cAMP-responsive transcription factor, CREB, is required for formation of long term memory (LTM) in Drosophila melanogaster and regulates transcription of a circadian clock gene, period (per). Involvement of CREB both in LTM and circadian rhythm raises the possibility that per also plays a role in LTM. Assaying the experience-dependent courtship inhibition in male flies as a measure for LTM, we show here that per mutants are defective in LTM formation. This defect was rescued by induction of a wild-type per transgene in a per-null mutant, and overexpression of per enhanced LTM formation in the wild-type background. Furthermore, we found that synaptic transmission through per-expressing cells is most likely to be required during retrieval of LTM. In contrast, mutations in other clock genes (timeless, dClock, and cycle) did not affect LTM formation. Thus, independent of the core oscillator of circadian clock, per plays a key role in LTM formation. PMID- 15522976 TI - Poisoning in older adults: a 5-year experience of US poison control centers. AB - BACKGROUND: Poisoning in older adults has received relatively little attention. OBJECTIVE: To describe poison exposures in older adults reported to US poison centers and identify substances that pose a unique risk to this population. METHODS: A retrospective review of human exposures for patients aged >or=60 years reported to the American Association of Poison Control Center's Toxic Exposure Surveillance System from 1993 to 1997 was performed. Frequencies and cross tabulations were used to describe the data. Hazard factor analysis was conducted to identify medications that pose risk in this population. Statistical analysis included chi(2) and Fisher's exact test for hazard factors. RESULTS: A total of 298,713 poison exposure cases were reported to US poison centers involving individuals aged >or=60 years between 1993 and 1997. The proportion of cases in this age group also steadily increased from 2.5% of all cases in 1993 to 3.4% of all cases in 1997. The mean age of the patients was 64.7 years, and 34.1% were males. Hazard factor analysis was conducted on unintentional exposures and adverse reactions to pharmaceutical agents to determine medications that posed increased risk to older adults. The substance categories that had the highest hazard factor were radiopharmaceuticals, asthma therapies, anti-coagulants, anesthetics, and antidepressants. CONCLUSIONS: Therapeutic errors and adverse reactions to medications are common reasons for major effects and fatal outcomes among older adults reported to poison centers. Understanding poisoning issues specific to this population may help direct future outreach education efforts. PMID- 15522977 TI - Intramuscular olanzapine in the management of acute agitation. AB - OBJECTIVE: To review the current efficacy and safety evidence for the use of intramuscular olanzapine in the management of acute agitation. DATA SOURCES: MEDLINE, EMBASE, and PubMed (all to March 2004) were searched for full-text, English-language publications in humans. Search terms included olanzapine, psychosis, agitation, psychiatric emergency, and intramuscular. STUDY SELECTION AND DATA EXTRACTION: Prospective, randomized, controlled trials that evaluated efficacy or safety endpoints of intramuscular olanzapine in the management of acute agitation were included. All studies were evaluated independently by both authors. For clinical outcomes (efficacy, safety), the definitions as specified by each study were used. DATA SYNTHESIS: Four prospective trials were included in this review. Intramuscular olanzapine is comparable to haloperidol or lorazepam monotherapy in managing acute agitation associated with schizophrenia and dementia. Intramuscular olanzapine is superior to lorazepam monotherapy in the management of agitation associated with bipolar affective disorder. Preliminary evidence demonstrates that intramuscular olanzapine is associated with fewer adverse movement disorders than monotherapy with intramuscular haloperidol. Interpretation of published evidence is limited by confounding factors of comparator regimens and the patient populations studied. CONCLUSIONS: Additional studies comparing intramuscular olanzapine with combination antipsychotic/benzodiazepine therapy in more severely ill patients and patients with concomitant medical illnesses are needed to determine the most effective dosing regimen, use of adjunctive medications, and to obtain a comprehensive safety profile. PMID- 15522978 TI - First patient-centered drug information service in Germany--a descriptive study. AB - BACKGROUND: The majority of patients have unmet drug information needs, and patients' knowledge of their own treatment is often poor. This is a major obstacle to a more patient-centered healthcare system. OBJECTIVE: To explore patterns of patients' unmet drug information needs and outcomes of counseling patients by a drug and therapy information center (DTIC). METHODS: We established the first German DTIC for patients. In this descriptive study, data obtained on callers within 24 months were analyzed. Questionnaires for gathering information about callers' characteristics and the outcome of the advice were sent to all patients using the service after consultation. Data on all inquiries and evaluation sheets were documented and analyzed by a standardized database. RESULTS: During 24 months, 2049 telephone calls were recorded. Patients' unmet information needs were mainly related to adverse drug reactions/drug interactions (31.0%) and therapy information (27.2%). In 81.0% of the cases, patients' uncertainties regarding their prescribed medications were reduced, 37.9% discussed the advice with their physician, and 18.3% reported a reduction of physician visits as a result of our advice. The patient-physician relationship remained mainly (70.6%) unaffected after our intervention. CONCLUSIONS: The DTIC is useful as a source of medical expert advice. The service can help to reduce medication problems based on inadequate information and therefore avoid unnecessary healthcare utilization. With the knowledge provided, patients can participate in decisions affecting their own health. This procedure encourages greater confidence in the merits of drug therapy. PMID- 15522979 TI - Long-acting injectable risperidone. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and safety profile of long-acting (LA) risperidone for the treatment of schizophrenia. DATA SOURCES: Information was selected from PubMed (1965-July 2004). Applicable scientific posters were also used. STUDY SELECTION AND DATA EXTRACTION: All published information on risperidone LA was considered. Material providing a comprehensive description was considered. DATA SYNTHESIS: Risperidone LA is the first long-acting, injectable atypical antipsychotic. It is dosed at 25 50 mg every 2 weeks. Adverse effects are similar to those seen with oral risperidone. A short-term study showed that risperidone LA is better than placebo in reducing the signs and symptoms of schizophrenia, and a long-term trial showed that stable schizophrenic patients can be switched from either oral or other injectable antipsychotic medications to risperidone LA. CONCLUSIONS: Risperidone LA is efficacious and safe in the treatment of schizophrenia. PMID- 15522980 TI - Duloxetine: a dual reuptake inhibitor. AB - OBJECTIVE: To review the pharmacology, pharmacokinetics, clinical efficacy, and safety profile of duloxetine for the treatment of major depressive disorder (MDD). DATA SOURCES: Searches using MEDLINE and PsycINFO were conducted (1966 to November 2003). STUDY SELECTION AND DATA EXTRACTION: All duloxetine MDD information gathered was considered. Articles containing comprehensive information regarding duloxetine use for MDD were evaluated. DATA SYNTHESIS: Duloxetine is a serotonin-norepinephrine reuptake inhibitor being considered for treatment of MDD and stress urinary incontinence. While approved dosing ranges have not yet been determined, studies support the efficacy and safety of 40-60 mg twice daily for the treatment of acute MDD. Adverse effects have been of mild to moderate severity and are considered to be transient. Cardiovascular effects (increased heart rate or blood pressure), while present, do not appear to be clinically significant. Overall, duloxetine appears to be well tolerated. CONCLUSIONS: Duloxetine is a safe and effective antidepressant. Approval of this agent provides another treatment option for the management of MDD. PMID- 15522981 TI - Initiating warfarin therapy: 5 mg versus 10 mg. AB - OBJECTIVE: To review the literature investigating initial dosing of warfarin at 5 or 10 mg for treatment of acute venous thromboembolism. DATA SOURCES: Articles were identified through searches of MEDLINE (1966-December 2003) using the key words warfarin, oral anticoagulation, warfarin dose, warfarin initiation, venous thromboembolism, and anticoagulation. Additional references were located through review of the bibliographies of the articles cited. STUDY SELECTION AND DATA EXTRACTION: Studies of the initial dosing of warfarin at 5 or 10 mg were evaluated and relevant information was included, as were those that identified known factors that influence the maintenance dose of warfarin. DATA SYNTHESIS: For the treatment of acute venous thromboembolism, warfarin dosing is often provider dependent. Until recently, studies suggested that 5 mg initiation was as effective as 10 mg, without increasing the risk of bleeding. However, the most recent study comparing a 5- versus 10-mg initial dosing nomogram supports an initial dose of 10 mg. These results should be interpreted with caution, however, since patients at high risk for bleeding were excluded from the study. Several patient-specific factors will affect the maintenance dose, guiding clinicians to start with lower (<5 mg) or higher (>5 mg) doses. CONCLUSIONS: Although recent evidence supports a 10-mg initiation nomogram, clinicians should consider patient specific factors prior to deciding an initial warfarin dose. If a 10-mg loading dose is utilized, strict compliance with the protocol is necessary. PMID- 15522982 TI - Probable ischemic colitis caused by pseudoephedrine with tramadol as a possible contributing factor. AB - OBJECTIVE: To report a case of acute self-limiting ischemic colitis in a patient who was self-medicating with a proprietary over-the-counter oral decongestant containing pseudoephedrine. CASE SUMMARY: A 46-year-old white man developed clinical, endoscopic, and histologic features of acute ischemic colitis after taking a proprietary oral decongestant containing pseudoephedrine 240 mg/day for one week. The total daily dose was at the upper limit of recommended doses for pseudoephedrine (as a single drug or in combination products). The patient was also taking tramadol 150 mg/day for chronic back pain. He made a complete recovery. There were no other explanations for the episode of ischemic colitis. DISCUSSION: An objective causality assessment based on the Naranjo probability scale revealed pseudoephedrine to be a probable cause of ischemic colitis in our patient. Pseudoephedrine occasionally causes vascular insufficiency due to intense vasoconstriction, even at standard doses. Although our patient was not taking an excessive dose of pseudoephedrine, it is possible that the concurrent use of pseudoephedrine and tramadol may have increased adrenergic vasoconstriction, predisposing to ischemic colitis. CONCLUSIONS: Prolonged or intensive use of medications containing pseudoephedrine should be avoided, and the package information should contain advice that the medication should be ceased if abdominal pain or other ischemic symptoms occur. PMID- 15522983 TI - Pharmacist involvement in primary care improves hypertensive patient clinical outcomes. AB - BACKGROUND: The practice of pharmaceutical care in primary care settings in Thailand is currently not generally accepted. OBJECTIVE: To evaluate the effect of pharmacist involvement in treatment with hypertensive patients in primary care settings. METHODS: The treatment objective was to stabilize the blood pressure (BP) of hypertensive patients in accordance with the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure guidelines. Patients were randomly assigned to a pharmacist-involved group (treatment) or a group with no pharmacist involvement (control). Pre- and post test BPs, tablet counts, lifestyle modifications, and pharmacists' recommendations were recorded. The 6-month study was carried out in Mahasarakham University pharmacy and 2 primary care units. Patients were monitored monthly by reviewing their medications and supported by providing pharmaceutical care and counseling. RESULTS: From a total of 235 patients, the treatment group (n = 118) had a significant reduction in both systolic (S) and diastolic (D) BP compared with the 117 patients of the control group (p = 0.037, 0.027, respectively). The 158 patients (76 treatment, 82 control) with BPs >or=140/90 mm Hg at the beginning of the study showed significant BP reductions (p = 0.002 SBP, 0.008 DBP). The proportion of 158 patients whose BP became stabilized was higher in the treatment group (p = 0.017). The treatment group showed significantly better adherence (p = 0.014) and exercise control (p = 0.012) at the end of the study. Physicians accepted 42.72% of medication modifications and 5.34% of the suggestions for additional investigations. CONCLUSIONS: Hypertensive patients who received pharmacist input achieved a significantly greater benefit in BP reduction, BP control, and improvement in adherence rate and lifestyle modification. PMID- 15522984 TI - Immunolocalization of NHE8 in rat kidney. AB - In situ hybridization studies demonstrated that Na+/H+ exchanger NHE8 is expressed in kidney proximal tubules. Although membrane fractionation studies suggested apical brush-border localization, precise membrane localization could not be definitively established. The goal of the present study was to develop isoform-specific NHE8 antibodies as a tool to directly establish the localization of NHE8 protein in the kidney by immunocytochemistry. Toward this goal, two sets of antibodies that label different NHE8 epitopes were developed. Monoclonal antibody 7A11 and polyclonal antibody Rab65 both specifically labeled NHE8 by Western blotting as well as by immunofluorescence microscopy. The immunolocalization pattern in the kidney seen with both antibodies was the same, thereby validating NHE8 specificity. In particular, NHE8 expression was observed on the apical brush-border membrane of all proximal tubules from S1 to S3. The most intense staining was evident in proximal tubules in the deeper cortex and medulla with a significant but somewhat weaker staining in superficial proximal tubules. Colocalization studies with gamma-glutamyltranspeptidase and megalin indicated expression of NHE8 on both the microvillar surface membrane and the coated-pit region of proximal tubule cells, suggesting that NHE8 may be subject to endocytic retrieval and recycling. Although colocalizing in the proximal tubule with NHE3, no significant alteration in NHE8 protein expression was evident in NHE3-null mice. We conclude that NHE8 is expressed on the apical brush border membrane of proximal tubule cells, where it may play a role in mediating or regulating ion transport in this nephron segment. PMID- 15522985 TI - Abnormal EGF-dependent regulation of sodium absorption in ARPKD collecting duct cells. AB - Amiloride-sensitive sodium entry, via the epithelial sodium channel (ENaC), is the rate-limiting step for Na+ absorption in kidney collecting ducts, and epidermal growth factor (EGF) inhibits Na+ transport and ENaC expression. A pathognomonic feature of polycystic kidney disease (PKD) is EGF receptor mislocalization to the apical plasma membrane and EGF/EGF receptor axis overactivity. Immunohistochemical and biochemical analysis revealed mislocalization of EGF receptor and excessive activation of the p42/44 extracellular signal-regulated protein kinase pathway (ERK1/2) in kidneys from cystic mice compared with noncystic littermates. Primary monolayer cultures of noncystic and cystic murine collecting duct principal cells were used to identify aberrant EGF-dependent ERK1/2 activation and regulation of Na+ transport associated with autosomal recessive PKD. Addition of EGF to the basolateral bathing solution of noncystic or cystic monolayers led to p42/44 phosphorylation and inhibition of Na+ transport (30-35%), whereas apical EGF was effective only in monolayers derived from cystic mice. p42/44 Phosphorylation and inhibition of Na+ transport were prevented by prior treatment of the cells with an ERK kinase inhibitor. Chronic treatment (24 h) of noncystic and cystic monolayers with basolateral EGF elicited sustained inhibition of Na+ absorption (50-55%) and a reduction in steady-state ENaC mRNA levels (50-75%). In contrast, addition of EGF to the apical bathing solution (24 h) had no effect in noncystic monolayers but led to inhibition of Na+ transport (50-60%) and decreased ENaC expression (45 60%) in cystic cells. Pretreatment of the monolayers with an ERK kinase inhibitor abolished the chronic effects of EGF on Na+ transport. The results of these studies reveal that the mislocalized apical EGF receptors are functionally coupled to the ERK pathway and that abnormal EGF-dependent regulation of ENaC function and expression may contribute to PKD pathophysiology. PMID- 15522986 TI - Renal effects of Tamm-Horsfall protein (uromodulin) deficiency in mice. AB - The Tamm-Horsfall protein (THP; uromodulin), the dominant protein in normal urine, is produced exclusively in the thick ascending limb of Henle's loop. THP mutations are associated with disease; however, the physiological role of THP remains obscure. We generated THP gene-deficient mice (THP -/-) and compared them with wild-type (WT) mice. THP -/- mice displayed anatomically normal kidneys. Steady-state electrolyte handling was not different between strains. Creatinine clearance was 63% lower in THP -/- than in WT mice (P < 0.05). Sucrose loading induced no changes between strains. However, water deprivation for 24 h decreased urine volume from 58 +/- 9 to 28 +/- 4 microl x g body wt(-1) x 24 h(-1) in WT mice (P < 0.05), whereas in THP -/- mice this decrease was less pronounced (57 +/ 4 to 41 +/- 5 microl x g body wt(-1) x 24 h(-1); P < 0.05), revealing significant interstrain difference (P < 0.05). We further used RT-PCR, Northern and Western blotting, and histochemistry to study renal transporters, channels, and regulatory systems under steady-state conditions. We found that major distal transporters were upregulated in THP -/- mice, whereas juxtaglomerular immunoreactive cyclooxygenase-2 (COX-2) and renin mRNA expression were both decreased in THP -/- compared with WT mice. These observations suggest that THP influences transporters in Henle's loop. The decreased COX-2 and renin levels may be related to an altered tubular salt load at the macula densa, whereas the increased expression of distal transporters may reflect compensatory mechanisms. Our data raise the hypothesis that THP plays an important regulatory role in the kidney. PMID- 15522987 TI - Development of water transport in the collecting duct. AB - The ability of the immature kidney to concentrate urine is lower than in adults. This can lead to severe water and electrolyte disorders, especially in premature babies. Resistance to AVP and lower tonicity of the medullary interstitium seem to be the major factors limiting urine concentration in newborns. AVP-stimulated cAMP generation is impaired. This is the result of inhibition of the production by PGE(2) acting through EP3 receptors and increased degradation by phosphodiesterase IV. The expression of aquaporin-2 (AQP2) in the immature kidney is low; however, under conditions of water deprivation and after stimulation with DDAVP, it rises to adult levels. The expression of AQP3 and AQP4 is intact at birth and does not seem to contribute to the hyporesponsiveness to AVP. Low sodium transport by thick ascending loops of Henle, immaturity of the medullary architecture, and adaptations in the transport of urea contribute to the lower tonicity of the medullary interstitium. This paper reviews the alterations in the AVP signal transduction pathway in the immature kidney. PMID- 15522988 TI - MAP kinases and the adaptive response to hypertonicity: functional preservation from yeast to mammals. AB - The adaptation to hypertonicity in mammalian cells is driven by multiple signaling pathways that include p38 kinase, Fyn, the catalytic subunit of PKA, ATM, and JNK2. In addition to the well-characterized tonicity enhancer (TonE) TonE binding protein interaction, other transcription factors (and their respective cis elements) can potentially respond to hypertonicity. This review summarizes the current knowledge about the signaling pathways that regulate the adaptive response to osmotic stress and discusses new insights from yeast that could be relevant to the osmostress response in mammals. PMID- 15522989 TI - Protection against proximal tubule necrosis with 11-deoxy-16,16-dimethyl prostaglandin E2 in vitro. PMID- 15522990 TI - Plasma S-nitrosothiols and chronic renal failure. PMID- 15522993 TI - Postexercise insulin sensitivity is not impaired after an overnight lipid infusion. AB - High plasma fatty acid availability and a positive energy balance in sedentary individuals reduce insulin sensitivity. This study's purpose was to determine whether high plasma fatty acid availability and systemic caloric excess after exercise also impair insulin sensitivity. On two separate occasions, seven nonobese women performed 90 min of exercise at approximately 65% peak oxygen uptake. In one trial, a lipid + heparin emulsion (Lipid) was infused overnight to increase plasma fatty acid availability. In the other trial, saline was infused as control. The next morning, a muscle biopsy was taken to measure muscle glycogen and intramuscular triglyceride (IMTG) concentrations. Three hours after the overnight infusion was stopped, insulin sensitivity was assessed with an intravenous glucose tolerance test, using minimal model analysis (Si). During the overnight infusions, plasma fatty acid concentration was approximately fourfold higher [means (SD): 0.84 (0.36) vs. 0.22 (0.09) mmol/l; P = 0.003], and the next morning IMTG concentration was approximately 30% greater [49.2 (6.6) vs. 38.3 (7.7) mmol/kg dry wt; P = 0.036] in Lipid compared with saline. However, muscle glycogen concentration was not different between trials (P = 0.82). Lipid caused a 24-h surplus of approximately 1100 kcal above energy balance (P = 0.00001), whereas energy balance was maintained in saline. Despite these differences in fatty acid and energy availability, Si the morning after exercise was not different between trials (P = 0.72). Thus insulin sensitivity the morning after a single exercise session was not reduced despite overnight exposure to a fourfold increase in plasma fatty acid concentration, elevated IMTG concentration, and systemic delivery of approximately 1,100-kcal excess. PMID- 15522992 TI - Interaction of a selective serotonin reuptake inhibitor with insulin in the control of hepatic glucose uptake in conscious dogs. AB - Whether hyperinsulinemia is required for stimulation of net hepatic glucose uptake (NHGU) by a selective serotonin reuptake inhibitor (SSRI) was examined in four groups of conscious 42-h-fasted dogs, using arteriovenous difference and tracer ([3-3H]glucose) techniques. Experiments consisted of equilibration (-120 to -30 min), basal (-30 to 0 min), and experimental periods (Exp; 0-240 min). During Exp, somatostatin, intraportal insulin [at basal (Ins groups) or 4-fold basal rates (INS groups)], basal intraportal glucagon, and peripheral glucose (to double hepatic glucose load) were infused. In the Fluv-Ins (n = 7) and Fluv-INS groups (n = 6), saline was infused intraportally from 0 to 90 min (P1), and fluvoxamine was infused intraportally at 2 microg x kg(-1) x min(-1) from 90 to 240 min (P2). Sal-Ins (n = 9) and Sal-INS (n = 8) received intraportal saline in P1 and P2. NHGU during P2 was 8.4 +/- 1.4 and 6.9 +/- 2.3 micromol x kg(-1) x min(-1) in Sal-Ins and Fluv-Ins, respectively (not significant), and 13.3 +/- 2.2 and 20.9 +/- 3.1 micromol x kg(-1) x min(-1) (P < 0.05) in Sal-INS and Fluv-INS. Unidirectional (tracer-determined) hepatic glucose uptake was twofold greater (P < 0.05) in Fluv-INS than Sal-INS. Net hepatic carbon retention during P2 was significantly greater in Fluv-INS than Sal-INS (18.5 +/- 2.7 vs. 12.2 +/- 1.9 micromol x kg(-1) x min(-1)). Nonhepatic glucose uptake was reduced in Fluv-INS vs. Sal-INS (20.0 +/- 1.3 vs. 38.4 +/- 5.4 micromol x kg(-1) x min(-1), P < 0.05). Intraportal fluvoxamine enhanced NHGU and net hepatic carbon retention in the presence of hyperinsulinemia but not euinsulinemia, suggesting that hepatocyte-targeted SSRIs may reduce postprandial hyperglycemia. PMID- 15522994 TI - Molecular evidence supporting the portal theory: a causative link between visceral adiposity and hepatic insulin resistance. AB - The mechanism by which increased central adiposity causes hepatic insulin resistance is unclear. The "portal hypothesis" implicates increased lipolytic activity in the visceral fat and therefore increased delivery of free fatty acids (FFA) to the liver, ultimately leading to liver insulin resistance. To test the portal hypothesis at the transcriptional level, we studied expression of several genes involved in glucose and lipid metabolism in the fat-fed dog model with visceral adiposity vs. controls (n = 6). Tissue samples were obtained from dogs after 12 wk of either moderate fat (42% calories from fat; n = 6) or control diet (35% calories from fat). Northern blot analysis revealed an increase in the ratio of visceral to subcutaneous (v/s ratio) mRNA expression of both lipoprotein lipase (LPL) and peroxisome proliferator-activated receptor-gamma (PPARgamma). In addition, the ratio for sterol regulatory element-binding transcription factor-1 (SREBP-1) tended to be higher in fat-fed dogs, suggesting enhanced lipid accumulation in the visceral fat depot. The v/s ratio of hormone-sensitive lipase (HSL) increased significantly, implicating a higher rate of lipolysis in visceral adipose despite hyperinsulinemia in obese dogs. In fat-fed dogs, liver SREBP-1 expression was increased significantly, with a tendency for increased fatty acid binding protein (FABP) expression. In addition, glucose-6-phosphatase (G-6-Pase) and phosphoenolpyruvate carboxykinase (PEPCK) increased significantly, consistent with enhanced gluconeogenesis. Liver triglyceride content was elevated 45% in fat fed animals vs. controls. Moreover, insulin receptor binding was 50% lower in fat fed dogs. Increased gene expression promoting lipid accumulation and lipolysis in visceral fat, as well as elevated rate-limiting gluconeogenic enzyme expression in the liver, is consistent with the portal theory. Further studies will need to be performed to determine whether FFA are involved directly in this pathway and whether other signals (either humoral and/or neural) may contribute to the development of hepatic insulin resistance observed with visceral obesity. PMID- 15522995 TI - Activation of ubiquitin-proteasome pathway is involved in skeletal muscle wasting in a rat model with biliary cirrhosis: potential role of TNF-alpha. AB - Hepatic cirrhosis is associated with negative nitrogen balance and loss of lean body mass. This study aimed to identify the specific proteolytic pathways activated in skeletal muscles of cirrhotic rats. TNF-alpha can stimulate muscle proteolysis; therefore, a potential relationship between TNF-alpha and muscle wasting in liver cirrhosis was also evaluated. Cirrhosis was induced by bile duct ligation (BDL) in male adult Sprague-Dawley rats. mRNA and protein levels of various targets were determined by RT-PCR and Western blotting, respectively. The proteolytic rate was measured ex vivo using isolated muscles. Compared with sham operated controls, BDL rats had an increased degradation rate of muscle proteins and enhanced gene expression of ubiquitin, 14-kDa ubiquitin carrier protein E2, and the proteasome subunits C2 and C8 (P < 0.01). The muscle protein levels of free ubiquitin and conjugated ubiquitin levels were also elevated (P < 0.01). However, there was no difference between the two groups with regard to cathepsin and calpain mRNA levels. Cirrhotic muscle TNF-alpha levels were increased and correlated positively with free and conjugated ubiquitin (P < 0.01). We conclude that the ubiquitin-proteasome system is involved in muscle wasting of rats with BDL-induced cirrhosis. TNF-alpha might play a role in mediating activation of this proteolytic pathway, probably through a local mechanism. PMID- 15522996 TI - Circulating resistin in lean, obese, and insulin-resistant mouse models: lack of association with insulinemia and glycemia. AB - Resistin is an adipocyte-secreted hormone proposed to link obesity with insulin resistance and diabetes, but no previous study has performed a joint quantitative evaluation of white adipose tissue (WAT) resistin mRNA expression and serum levels in relation to insulinemia and glycemia in mice. We have thus comparatively assessed WAT resistin mRNA expression and serum resistin levels in lean C57BL/6J mice and various mouse models of obesity, including diet-induced obese (DIO) C57BL/6J mice, high fat-fed TNF-alpha-/- mice, and brown adipose tissue (BAT)-deficient uncoupling protein-diphtheria toxin A chain (UCP1-DTA) mice. We also studied whether treatment with the weight-reducing and insulin sensitizing compounds, MTII, an alpha-melanocyte-stimulating hormone analog, or CNTF(Ax15), a ciliary neurotrophic factor analog, alters resistin mRNA expression and/or circulating levels in lean and DIO C57BL/6J mice. We find that resistin mRNA expression is similar in DIO and lean C57BL/6J mice, as well as in TNF-alpha /- and wild-type (WT) mice. Circulating resistin levels, however, are higher in DIO C57BL/6J, high fat-fed TNF-alpha-/-, and UCP1-DTA mice compared with lean controls. Moreover, although resistin mRNA expression is upregulated by MTII treatment for 24 h and downregulated by CNTF(Ax15) treatment for 3 or 7 days, circulating resistin levels are not altered by MTII or CNTF(Ax15) treatment. In addition, serum resistin levels, but not resistin mRNA expression levels, are correlated with body weight, and neither resistin mRNA expression nor serum resistin levels are correlated with serum insulin or glucose levels. We conclude that transcriptional regulation of resistin in WAT does not correlate with circulating resistin levels and that circulating resistin is unlikely to play a major endocrine role in insulin resistance or glycemia in mice. PMID- 15522997 TI - Characterization of beta-cell function impairment in first-degree relatives of type 2 diabetic subjects: modeling analysis of 24-h triple-meal tests. AB - To investigate early secretory defects in prediabetes, we evaluated beta-Cell function and insulin sensitivity (M value, by euglycemic clamp) in 26 normotolerant first-degree relatives of type 2 diabetic patients (FDR) and 17 age and weight-matched control subjects. beta-Cell function was assessed by modeling analysis of glucose and C-peptide concentrations measured during 24 h of standardized living conditions. Fasting and total insulin secretion (ISR) were increased in FDR, as was ISR at a reference 5 mM glucose level (ISR5, 107 +/- 6 vs. 87 +/- 6 pmol x min(-1) x m(-2), P < 0.05). ISR5 was inversely related to M in controls (ISR5 = k/M1.23, rho = -0.74, P < 0.005) but not in FDR; when M was accounted for (by calculating a compensation index ISR5 x M1.23), compensation for insulin resistance was impaired in FDR (10.8 +/- 1.0 vs. 13.4 +/- 0.6 units, P < 0.05). Potentiation of ISR, expressing relative transient increases in glucose-stimulated ISR during meals, was impaired in FDR (1.29 +/- 0.08 vs. 1.62 +/- 0.08 during 1st meal, P < 0.02). Moreover, the potentiation time course was related to glucose-dependent insulin-releasing polypeptide (GIP) concentrations in both groups, and the sensitivity of potentiation to GIP derived from this relationship tended to be impaired in FDR. Compensation index, potentiation, and sensitivity to GIP were interrelated parameters (P < 0.05 or less). beta-Cell function parameters were also related to mean 24-h glucose levels (r2 = 0.63, P < 0.0001, multivariate model). In conclusion, although in absolute terms ISR is increased in insulin-resistant FDR, beta-cell function shows a cluster of interrelated abnormalities involving compensation for insulin resistance, potentiation, and sensitivity to GIP, suggesting a beta-cell defect in the amplifying pathway of insulin secretion. PMID- 15522998 TI - Structural and functional analysis of pancreatic islets preserved by pioglitazone in db/db mice. AB - To evaluate preventive effects of pioglitazone on pancreatic beta-cell damage in C57BL/KsJ db/db mice, an obese diabetic animal model, the pancreatic islets were compared morphologically between pioglitazone-treated (100 mg/kg daily po) and untreated db/db mice (n = 7 for each) after a 12-wk intervention (6-18 wk of age). The fasting blood glucose level was significantly improved by the treatment with pioglitazone (260 +/- 12 vs. 554 +/- 62 mg/dl, P < 0.05). The islet mass in the pancreas was significantly greater in pioglitazone-treated mice than in untreated mice (10.2 +/- 1.1 vs. 4.6 +/- 0.2 mg, P < 0.01). Subsequently, biochemical and physiological analyses of the beta-cell function were employed using pioglitazone-treated and untreated db/db mice (n = 6 for each) and pioglitazone-treated and untreated db/+ mice (n = 6 for each). After 2 wk of treatment (10-12 wk of age), the plasma levels of triglyceride and free fatty acid were significantly decreased, whereas the plasma adiponectin level increased significantly compared with the untreated group (65.2 +/- 18.0 vs. 18.3 +/- 1.3 microg/ml, P < 0.05). Pioglitazone significantly reduced the triglyceride content in the islets (43.3 +/- 3.6 vs. 65.6 +/- 7.6 ng/islet, P < 0.05) with improved glucose-stimulated insulin secretion. Pioglitazone showed no significant effects on the biochemical and physiological parameters in db/+ mice. The present study first demonstrated that pioglitazone prevents beta-cell damage in an early stage of the disease progression in db/db mice morphologically and physiologically. Our results suggest that pioglitazone improves glucolipotoxicity by increasing insulin sensitivity and reducing fat accumulation in the pancreatic islets. PMID- 15522999 TI - Memantine blocks alpha7* nicotinic acetylcholine receptors more potently than n methyl-D-aspartate receptors in rat hippocampal neurons. AB - The N-methyl-d-aspartate (NMDA) receptor antagonist memantine is an approved drug for treatment of Alzheimer's disease (AD). Other such treatments are cholinesterase inhibitors and nicotinic acetylcholine receptor (nAChR) sensitizing agents such as galantamine. The present study was designed to test whether memantine exerts any effect on the cholinergic system, in particular the Ca(2+)-conducting alpha7(*) nAChR, in cultured hippocampal neurons. Memantine caused a concentration-dependent reduction of the amplitudes of whole-cell currents evoked by the alpha7(*) nAChR-selective agonist choline (10 mM) or by N methyl-d-aspartate (NMDA) (50 muM) plus glycine (10 muM). It also inhibited tonically activated NMDA receptors. Memantine was more potent in inhibiting alpha7(*) nAChRs than NMDA receptors; at -60 mV, the IC(50) values for memantine were 0.34 and 5.1 muM, respectively. Consistent with an open-channel blocking mechanism, memantine-induced NMDA receptor inhibition was voltage and use dependent; the Hill coefficient (n(H)) was approximately 1. Memantine-induced alpha7(*) nAChR inhibition had an n(H) < 1 and showed a variable voltage dependence; the effect was voltage-independent at 0.1 muM, becoming voltage dependent at >/=1 muM. Thus, memantine interacts with more than one class of sites on the alpha7(*) nAChRs. One is voltage-sensitive and therefore likely to be within the receptor channel. The other is voltage-insensitive and therefore likely to be in the extracellular domain of the receptor. It is suggested that blockade of alpha7(*) nAChRs by memantine could decrease its effectiveness for treatment of AD, particularly at early stages when the degrees of nAChR dysfunction and of cognitive decline correlate well. PMID- 15523000 TI - Poly(ADP-Ribose) polymerase promotes cardiac remodeling, contractile failure, and translocation of apoptosis-inducing factor in a murine experimental model of aortic banding and heart failure. AB - Oxidant stress-induced activation of poly(ADP-ribose) polymerase (PARP) plays a role in the pathogenesis of various cardiovascular diseases. We have now investigated the role of PARP in the process of cardiac remodeling and heart failure in a mouse model of heart failure induced by transverse aortic constriction (banding). The catalytic activity of PARP was inhibited by the potent isoindolinone-based PARP inhibitor INO-1001 or by PARP-1 genetic deficiency. PARP inhibition prevented the pressure overload-induced decrease in cardiac contractile function, despite the pressure gradient between both carotid arteries being comparable in the two experimental groups. The development of hypertrophy, the formation of collagen in the hearts, and the mitochondrial-to nuclear translocation of the cell death factor apoptosis-inducing factor (AIF) were attenuated by PARP inhibition. The ability of the inhibitor to block the catalytic activity of PARP was confirmed by immunohistochemical detection of poly(ADP-ribose), the product of the enzyme in the heart. Plasma levels of INO 1001, as measured at the end of the experiments, were in the concentration range sufficient to block the oxidant-mediated activation of PARP in murine cardiac myocytes in vitro. Myocardial hypertrophy and AIF translocation was also reduced in PARP-1-deficient mice undergoing aortic banding, compared with their wild-type counterparts. Overall, the current results demonstrate the importance of poly(ADP ribos)ylation in the pathogenesis of banding-induced heart failure. PMID- 15523001 TI - The selective alpha7 nicotinic acetylcholine receptor agonist PNU-282987 [N-[(3R) 1-Azabicyclo[2.2.2]oct-3-yl]-4-chlorobenzamide hydrochloride] enhances GABAergic synaptic activity in brain slices and restores auditory gating deficits in anesthetized rats. AB - Schizophrenic patients are thought to have an impaired ability to process sensory information. This deficit leads to disrupted auditory gating measured electrophysiologically as a reduced suppression of the second of paired auditoryevoked responses (P50) and is proposed to be associated with decreased function and/or expression of the homomeric alpha7 nicotinic acetylcholine receptor (nAChR). Here, we provide evidence that N-[(3R)-1-azabicyclo[2.2.2]oct-3 yl]-4-chlorobenzamide hydrochloride (PNU-282987), a novel selective agonist of the alpha7 nAChR, evoked whole-cell currents from cultured rat hippocampal neurons that were sensitive to the selective alpha7 nAChR antagonist methyllycaconitine (MLA) and enhanced GABAergic synaptic activity when applied to hippocampal slices. Amphetamine-induced sensory gating deficit, determined by auditory-evoked potentials in hippocampal CA3 region, was restored by systemic administration of PNU-282987 in chloral hydrate-anesthetized rats. Auditory gating of rat reticular thalamic neurons was also disrupted by amphetamine; however, PNU-282987 normalized gating deficit only in a subset of tested neurons (6 of 11). Furthermore, PNU-282987 improved the inherent hippocampal gating deficit occurring in a subpopulation of anesthetized rats, and enhanced amphetamine-induced hippocampal oscillation. We propose that the alpha7 nAChR agonist PNU-282987, via modulating/enhancing hippocampal GABAergic neurotransmission, improves auditory gating and enhances hippocampal oscillatory activity. These results provide further support for the concept that drugs that selectively activate alpha7 nAChRs may offer a novel, potential pharmacotherapy in treatment of schizophrenia. PMID- 15523002 TI - Ciglitazone-induced lenticular opacities in rats: in vivo and whole lens explant culture evaluation. AB - The cataractogenic potential of the thiazolidinedione ciglitazone (CIG) was investigated in vivo and in vitro. In the rat, CIG caused a dose-dependent (30 300 mg/kg/day) increase in incidence and severity of nuclear cataract formation during a 3-month nonclinical safety assessment study. Potential mechanisms of toxicity were surveyed using whole rat lens explants exposed to CIG with or without various inhibitors of cataract formation. In vitro, CIG caused a concentration-(0.375-30 muM) and time-dependent (3-24 h) change in biochemical [ATP content or mitochondrial reduction of the tetrazolium dye 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) and reduced glutathione (GSH) content] and morphometric (lens wet weight and clarity) markers of damage. Within 3 h of exposure, 7.5 muM CIG decreased lens ATP content 37 +/- 7% (percentage of difference from control, p < 0.05). After 24 h of exposure, lens ATP content, MTT reduction, and GSH content declined 57 +/- 5, 30 +/- 28, and 42 +/- 8%, respectively. Lens wet weight increased 17 +/- 4% with a concomitant decrement in lens clarity. Pretreating lenses with the mitochondrial calcium uniport inhibitor ruthenium red (RR) partially or fully protected lenses from toxicity. In contrast, the antioxidant dithiothreitol, aldose reductase inhibitor sorbinil, and selective cell-permeable calpain inhibitors [calpain II inhibitor and (2S,3S) trans-epoxysuccinyl-l-leucylamido-3-methylbutane ethyl ester (E64d)] were ineffective in providing protection under the present testing conditions. Early and selective changes in lenticular ATP content and the partial or full protective effect of RR suggest that alterations in lens bioenergetics may play an important role in CIG-induced cataract formation. Lens explant cultures were successfully used to select two thiazolidinediones that lacked cataractogenic activity when evaluated in 3-month rat safety assessment studies. PMID- 15523003 TI - Mechanism-based inactivation of CYP3A by HIV protease inhibitors. AB - Human immunodeficiency virus (HIV) protease inhibitors (PIs) are inhibitors of CYP3A enzymes, but the mechanism is poorly defined. In this study, time- and concentration-dependent decreases in activity as defined by maximum rate of inactivation (k(inact)) and inhibitor concentration that gives 50% maximal inactivation (K(I)) of CYP3A by amprenavir, indinavir, lopinavir, nelfinavir, ritonavir, and saquinavir were quantified using testosterone 6beta-hydroxylation as a marker for CYP3A activity with recombinant CYP3A4(+b(5)), recombinant CYP3A5, and pooled human liver microsomes (HLMs). All the PIs, except indinavir, displayed inactivation with CYP3A4(+b(5)) and HLMs. Ritonavir was the most potent (K(I) = 0.10 and 0.17 microM) and demonstrated high k(inact) values (0.32 and 0.40 min(-1)) with both CYP3A4(+b(5)) and HLMs. Ritonavir was not significantly depleted by high-affinity binding with CYP3A4(+b(5)) and confirmed that estimation of reversible inhibition was confounded with irreversible inhibition. For CYP3A5, nelfinavir exhibited the highest k(inact) (0.47 min(-1)), but ritonavir was the most potent (K(I) = 0.12 microM). Saquinavir and indinavir did not show time- and concentration-dependent decreases in activity with CYP3A5. Spectrophototmetrically determined metabolic intermediate complex formation was observed for all of the PIs with CYP3A4(+b(5)), except for lopinavir and saquinavir. The addition of nucleophilic and free aldehyde trapping agents and free iron and reactive oxygen species scavengers did not prevent inactivation of CYP3A4(+b(5)) by ritonavir, amprenavir, or nelfinavir, but glutathione decreased the inactivation by saquinavir (17%) and catalase decreased the inactivation by lopinavir (39%). In conclusion, all the PIs exhibited mechanism-based inactivation, and predictions of the extent and time course of drug interactions with PIs could be underestimated if based solely on reversible inhibition. PMID- 15523004 TI - Proximal row carpectomy: study with a minimum of ten years of follow-up. AB - BACKGROUND: Proximal row carpectomy is an accepted motion-sparing surgical procedure for the treatment of degenerative conditions of the wrist. However, there is little information regarding the long-term clinical and radiographic results following this procedure. METHODS: Twenty-two wrists in twenty-one patients underwent proximal row carpectomy for the treatment of degenerative arthritis between 1980 and 1992. Objective and subjective function was assessed after a minimum duration of follow-up of ten years (average, fourteen years). RESULTS: There were four failures (18%) requiring fusion at an average of seven years. All four failures occurred in patients who were thirty-five years of age or less at the time of the proximal row carpectomy (p = 0.03). The wrists that did not fail had an average flexion-extension arc of 72 degrees , associated with an average grip strength of 91% of that on the contralateral side. The patients were very satisfied with fourteen of the eighteen wrists that did not fail and were satisfied with the remaining four. The patients rated nine wrists as not painful, four as mildly painful, five as moderately painful, and none as severely painful. The average Disabilities of the Arm, Shoulder and Hand score was 9 points. Radiographs revealed no loss of the radiocapitate space in three of the seventeen wrists for which radiographs were made, reduced space in seven, and complete loss of the space in seven. With the numbers available, there was no significant association between loss of joint space seen on radiographs and subjective and objective function. CONCLUSIONS: At the time of long-term follow up, all patients older than thirty-five years of age at the time of a proximal row carpectomy had maintained a satisfactory range of motion, grip strength, and pain relief and were satisfied with the result. Caution should be exercised in performing the procedure in patients younger than thirty-five years of age. Although degeneration of the radiocapitate joint was seen radiographically in fourteen of the seventeen wrists, it did not preclude a successful clinical result. PMID- 15523005 TI - The epidemiology of posttraumatic adult respiratory distress syndrome. AB - BACKGROUND: Although adult respiratory distress syndrome is an important early complication of blunt trauma, the epidemiology and risk factors for its development remain poorly defined. The aims of this study were to determine the prevalence and demographics of this complication in a prospective cohort series of patients admitted to the hospital following injury. We also assessed the contribution of the severity and pattern of the injury to the risk of this complication developing. By identifying factors associated with the highest risk of the development of adult respiratory distress syndrome, we aimed to produce guidelines to facilitate earlier detection. METHODS: We prospectively studied 7192 patients admitted to a single university hospital, over an eight-year period, for treatment of a traumatic injury. With the exception of patients who had sustained a hip fracture or who had been discharged within seventy-two hours after admission, all patients who required hospital admission following trauma, were older than thirteen years of age, and were a resident within the catchment area were included in the analysis. The prevalence and demographics of posttraumatic adult respiratory distress syndrome were identified for patients who had sustained musculoskeletal, thoracic, abdominal, and head injuries, either in isolation or in combination. The relative risks of this condition developing were calculated according to the injury pattern. Multiple logistic regression analysis was performed to identify the most highly significant predictors of the development of adult respiratory distress syndrome. RESULTS: Adult respiratory distress syndrome developed in thirty-six (0.5%) of the patients. The prevalence was significantly higher among younger patients (p = 0.002), and 83% of the cases followed high-energy trauma. The prevalence of adult respiratory distress syndrome after isolated thoracic, head, abdominal, or extremity injury was <1%. Patients with injuries to two anatomical regions had a higher prevalence (up to 2.9%), and those with injuries to three anatomical regions had an even higher prevalence (up to 10.2%). Multiple logistic regression analysis showed the Injury Severity Score, the presence of a femoral fracture, the combination of abdominal and extremity injuries, and observations of compromised physiological function on admission each to be an independent predictor of the later development of adult respiratory distress syndrome. CONCLUSIONS: The prevalence of adult respiratory distress syndrome increases with injury severity and combinations of injuries to more than one anatomical region. We have been able to quantify the importance and relative risks associated with these injuries. The implications of our findings with regard to facilitating early detection of this complication are discussed. PMID- 15523006 TI - Botulinum toxin as an adjunct to serial casting treatment in children with cerebral palsy. AB - BACKGROUND: Although botulinum toxin A is frequently used to augment serial casting in the treatment of soft-tissue contractures in children with cerebral palsy, its effectiveness for this purpose has not been evaluated. The purpose of the present study was to determine whether botulinum toxin A injection increases the efficacy of serial casting. METHODS: A prospective, randomized trial was undertaken to compare serial casting only with serial casting combined with botulinum toxin A (Botox) injection for the treatment of ankle equinus contractures in twenty-three children with cerebral palsy. Range-of-motion testing, spasticity assessment, and computerized gait analysis were performed as long as twelve months after treatment. RESULTS: There was no difference between the groups with regard to the duration of casting required to correct the equinus contracture. Both groups maintained a significant improvement in passive ankle dorsiflexion throughout the follow-up period, although the group managed with casting and Botox had a significant loss of dorsiflexion when the values at six, nine, and twelve months were compared with the value at three months. Peak dorsiflexion during the stance and swing phases was significantly improved in both groups at three months but only in the group managed with casting alone at twelve months. Plantar flexor spasticity was significantly decreased at three months in both groups, but it was significantly decreased at six, nine, and twelve months only in the group managed with casting alone. Spasticity was significantly greater in the group managed with casting and Botox than it was in the group managed with casting only at six, nine, and twelve months. CONCLUSIONS: The present study demonstrates the efficacy of serial casting in the treatment of equinus contractures in children with cerebral palsy who are able to walk. Contrary to our hypothesis, the addition of botulinum toxin A to a serial casting regimen led to earlier recurrence of spasticity, contracture, and equinus during gait. The results of the present study suggest that botulinum toxin combined with serial casting for the treatment of fixed contractures will lead to a recurrence of plantar flexor spasticity and equinus contracture by six months in this patient population. While previous research has indicated that the injection of botulinum toxin A is superior to casting for the treatment of dynamic equinus, the present study suggests that serial casting alone is preferable for the treatment of fixed equinus contractures in children with cerebral palsy. PMID- 15523007 TI - Acetabular reconstruction with impaction bone-grafting and a cemented cup in patients younger than fifty years old. AB - BACKGROUND: Acetabular bone deficiency can present a challenge during total hip arthroplasty, especially in young patients. The purpose of the present study was to evaluate the long-term clinical and radiographic outcomes of primary and revision acetabular reconstruction with use of an impaction bone-grafting technique and a cemented polyethylene cup in young patients who had preexisting acetabular bone deficiency. METHODS: Forty-two consecutive acetabular reconstructions were performed in thirty-seven patients who were younger than fifty years old (average, 37.2 years old). The impaction bone-grafting technique was used for twenty-three primary and nineteen revision acetabular reconstructions. Twenty-eight patients (thirty-one hips) were available for review after a minimum duration of follow-up of fifteen years. Clinical and radiographic results were assessed, and survivorship analysis was performed with the Kaplan-Meier method. RESULTS: Eight hips were revised at a mean of twelve years (range, three to twenty-one years) after a primary reconstruction (four hips) or revision reconstruction (four hips). The revision was performed because of aseptic loosening of the acetabular component in four hips and because of culture-proven septic loosening in two. Two additional cups (both in hips that had had a revision reconstruction) were revised, during revision of the femoral stem, because of wear (one hip) or because of persistent intraoperative instability (one hip). Twenty-eight hips (in twenty-five patients) had retention of the acetabular component for a minimum of fifteen years. The mean Harris hip score for that group was 89 points. Twenty-six of these twenty-eight hips had no or slight pain. Kaplan-Meier analysis revealed a twenty-year survival rate of 80% (95% confidence interval, 67% to 94%) with acetabular revision for any reason as the end point and of 91% (95% confidence interval, 80% to 100%) with acetabular revision because of aseptic loosening as the end point. CONCLUSIONS: Acetabular reconstruction with use of impaction bone-grafting and a cemented polyethylene cup is a reliable and durable technique that is associated with good long-term results in young patients with acetabular bone-stock defects. PMID- 15523008 TI - Ankle stress test for predicting the need for surgical fixation of isolated fibular fractures. AB - BACKGROUND: The purpose of this study was to confirm the prevalence of medial ankle widening among patients with an isolated fibular fracture and to determine the functional outcome of nonoperative treatment despite a diagnosis of a supination-external rotation stage-IV injury based on stress radiography. METHODS: One hundred and one patients with evidence of an isolated fibular fracture and an intact mortise seen on a standard ankle trauma radiograph series were evaluated with stress radiographs. Clinical signs were recorded at the time of presentation. A positive stress test was defined as > or =4 mm of widening of the medial clear space. Patients with a negative stress test were treated nonoperatively, those with a positive stress test and clinical signs of medial injury were treated surgically, and those with a positive stress test and no signs of medial injury were treated according to the preference of the surgeon and patient. The patients were followed prospectively with radiographs and ankle outcome scores. RESULTS: Sixty-six (65%) of the 101 patients had a positive stress radiograph. Thirty-six of them had signs of medial injury, and thirty had no medial injury. With regard to predicting a positive stress radiograph, medial tenderness had a sensitivity of 56% and a specificity of 80%, swelling had a sensitivity of 55% and a specificity of 71%, and ecchymosis had a sensitivity of 26% and a specificity of 91%. Of the subset of patients without signs of medial injury, twenty were treated nonoperatively (group I) and ten were treated operatively (group II). Two of the twenty patients in group I had evidence of persistent widening of the medial clear space at the time of the latest follow-up (mean, 7.4 months); only one of those patients was symptomatic. The average American Orthopaedic Foot and Ankle Society (AOFAS) score was 94 points in group I and 93 points in group II. CONCLUSIONS: We found a high rate of positive stress radiographs for patients who presented with an isolated fibular fracture and an intact ankle mortise on the initial radiographs. Medial tenderness, swelling, and ecchymosis were not sensitive with regard to predicting widening of the medial clear space on stress radiographs. All of the patients with a positive stress radiograph and no clinical symptoms who were treated without surgery had a good or excellent clinical result. PMID- 15523009 TI - Low-intensity pulsed ultrasound accelerates maturation of callus in patients treated with opening-wedge high tibial osteotomy by hemicallotasis. AB - BACKGROUND: Opening-wedge high tibial osteotomy by hemicallotasis for osteoarthritis in the medial compartment of the knee requires external fixation for a long time, until callus maturation is complete. The aim of this study was to determine if low-intensity pulsed ultrasound would accelerate callus maturation when applied after distraction to limbs treated with opening-wedge high tibial osteotomy by hemicallotasis. METHODS: Twenty-one patients with symmetric grades of osteoarthritis and similar degrees of varus deformity in the two knees underwent bilateral one-stage opening-wedge high tibial osteotomy by hemicallotasis. After completion of distraction, the bone mineral density of the distraction callus was measured. Then, one randomly selected limb was subjected to ultrasound treatment for twenty minutes daily until removal of the external fixator. The contralateral limb was left untreated to serve as the control. After four weeks of treatment, bone mineral density was measured again. RESULTS: During the four-week treatment period, the mean increase in callus bone mineral density was significantly greater in the ultrasound-treated tibiae (0.20 +/- 0.12 g/cm(2)) than in the control tibiae (0.13 +/- 0.10 g/cm(2)) (p = 0.02, unpaired t test). In eighteen patients the increase in the bone mineral density was greater in the ultrasound-treated limb than in the control limb, whereas in three patients the increase was greater in the control limb. CONCLUSIONS: We found that low-intensity pulsed ultrasound applied during the consolidation phase of distraction osteogenesis accelerates callus maturation after opening-wedge high tibial osteotomy by hemicallotasis in elderly patients. PMID- 15523010 TI - Complications encountered during lengthening over an intramedullary nail. AB - BACKGROUND: In limb-lengthening, the quest for increased patient comfort and a reduced period of external fixation has led to techniques such as lengthening over an intramedullary nail. The goals of this study were to investigate the rate and types of complications encountered during lengthening over an intramedullary nail and to identify solutions to these complications. METHODS: Forty-two segments (thirty-five femora and seven tibiae) in thirty-five patients were lengthened. The mean age of the patients was 26.6 years, the mean amount lengthened was 6.3 cm (range, 2.5 to 11.5 cm), the mean external fixation index was 18.7 days/cm, and the mean lengthening index was 31.2 days/cm. The patients were followed for a mean period of forty-four months postoperatively. RESULTS: Eighteen complications occurred in sixteen (38%) of the forty-two segments for a rate of 0.43 complication per segment. Complications were classified, according to the system of Paley et al., as two problems, thirteen obstacles, and three sequelae. Sixteen of them required additional surgical interventions. A preoperative score of >6.5 on the system of Paley et al., a lengthening of >6 cm, and a lengthening percentage of >21.5% of the original bone length were indicators of a higher probability of the occurrence of complications. CONCLUSIONS: Lengthening over an intramedullary nail provides increased patient comfort and reduces the external fixation period. If the problems encountered are treated aggressively, the result of the treatment can be quite satisfactory. PMID- 15523011 TI - Dedifferentiated chondrosarcoma: the role of chemotherapy with updated outcomes. AB - BACKGROUND: There are very few published data on the survival of patients with dedifferentiated chondrosarcoma, or, more specifically, on the efficacy and role of chemotherapy, especially in the era of modern diagnostic and treatment modalities. The current study examines the influence of advancements in imaging and chemotherapy on outcome and serves as an extension to a previous study published in 1986. METHODS: Forty-two patients with dedifferentiated chondrosarcoma who had presented to our institution between 1986 and 2000 were identified, and a retrospective chart review was performed. The study group included twenty-four men and eighteen women with an average age of sixty-six years. The diagnosis of dedifferentiated chondrosarcoma was verified histologically, and data on treatment, adjuvant therapy, and survival were obtained from the medical records of all patients. All patients had been followed for a minimum of twenty-four months. RESULTS: The tumors were classified, according to the system of the Musculoskeletal Tumor Society, as grade IIA (five), grade IIB (twenty-six), and grade III (eleven). Three patients underwent biopsy only, eighteen had a limb-sacrificing procedure, and twenty-one had a limb sparing procedure. In the group of patients who underwent resection, the surgical margins were classified as intralesional in three, marginal in two, wide in nineteen, and radical in fifteen. Twenty-seven patients received neoadjuvant therapy; of these, twenty-three received chemotherapy only, two received radiotherapy only, and two received combined therapy. The median survival time was 7.5 months, and the five-year rate of disease-free survival was 7.1%. With the numbers available, there was no significant difference in the rate of disease free survival with respect to the use of chemotherapy (p = 0.54), the location of surgical margins (p = 0.14), the histological subtype (p = 0.87), the tumor stage at the time of diagnosis (p = 0.43), the tumor size (p = 0.79), or the performance of limb-sparing as opposed to limb-sacrificing procedures (p = 0.42). CONCLUSIONS: Despite advances in diagnostic modalities and adjuvant therapies, dedifferentiated chondrosarcoma continues to carry a poor prognosis. While local control is achieved in the majority of cases, distant disease remains the greatest clinical challenge, developing in 90% of patients. Efforts are needed to continue to encourage earlier diagnosis and to develop effective adjuvant therapies for the control of distant disease. The routine use of current adjuvant chemotherapy and its inherent risks in this population should be reconsidered. PMID- 15523012 TI - Salvage of a recurrently dislocating total hip prosthesis with use of a constrained acetabular component. A concise follow-up of a previous report. AB - We previously reported the results at a mean of five years following the use of a tripolar constrained acetabular component to treat recurrently dislocating total hip prostheses. In this study, we report the results after longer follow-up, with emphasis on the prevalence of implant loosening, osteolysis, and later recurrent instability. Fifty-five patients treated with a total of fifty-six constrained acetabular components because of recurrent dislocations of a total hip prosthesis (average, six dislocations; range, two to twenty dislocations) were followed for an average of 10.2 years (range, 7.0 to 13.2 years) or until death. Four (7%) of the fifty-six hips had a subsequent dislocation or failure of the device. Three femoral components (5%) and two acetabular components (4%) were revised because of aseptic loosening. One hip was revised because of osteolysis. We concluded that this constrained acetabular component provides durable protection against additional dislocations without substantial deleterious effects on component fixation. We use this device to treat recurrent dislocation when other modalities are unlikely to be effective. PMID- 15523013 TI - Medial collateral ligament strain with partial posteromedial olecranon resection. A biomechanical study. AB - BACKGROUND: Partial resection of the posteromedial aspect of the olecranon in the treatment of valgus extension impingement osteophytosis is a well-described technique. It has been hypothesized that removal of the normal olecranon process, beyond the osteophytic margin, increases the strain in the anterior bundle of the medial collateral ligament. METHODS: We used an electromagnetic tracking device to investigate the strain in the anterior bundle of the medial collateral ligament as a function of increasing applied torque and posteromedial resections of the olecranon in seven cadaveric elbows. Applied torques under valgus stress consisted of hand weight, hand weight plus 1.75 Nm, and hand weight plus 3.5 Nm. Resections were conducted in sequential 3-mm increments, from 0 to 9 mm. We measured changes in the length of the anterior and posterior bands of the anterior bundle of the medial collateral ligament with strain gauges. The strains of the two bands were averaged, and the average was reported. RESULTS: The strain in the anterior bundle of the medial collateral ligament was found to increase with increasing flexion angle, valgus torque, and olecranon resection beyond 3 mm. In two elbows, the anterior bundle of the medial collateral ligament ruptured during testing following the 9-mm resection. There was a significant difference between the strain following the 6-mm resection and that following the 3-mm resection at 110 degrees of flexion with 3.5 Nm of added torque (p = 0.004). CONCLUSIONS: In this in vitro cadaver study, an increase in flexion angle, an increase in valgus torque, and resection of > or =6 mm led to an increase in strain in the anterior bundle of the medial collateral ligament. The non-uniform change in strain related to 3 mm of resection suggests that resections of the posteromedial aspect of the olecranon of >3 mm may jeopardize the function of the anterior bundle. PMID- 15523014 TI - The external rotation method for reduction of acute anterior dislocations and fracture-dislocations of the shoulder. AB - BACKGROUND: Several methods of reducing an acute anterior dislocation of the shoulder have been described. The aim of this study was to assess the effectiveness of the external rotation method in the reduction of acute anterior shoulder dislocations with and without fractures of the greater tuberosity and to evaluate the causes of failure. METHODS: Senior and junior orthopaedic residents attending in the Emergency Department were instructed in the external rotation method for the reduction of a shoulder dislocation in a classroom setting. Forty patients with an acute anterior dislocation of the shoulder, with or without an associated fracture of the greater tuberosity, who were treated with this method were evaluated prospectively. Data sheets completed by the orthopaedic residents when this method was used were evaluated with regard to the type of dislocation, the effectiveness of the procedure in achieving reduction, the need for premedication, the ease of performing the reduction, and complications, if any. RESULTS: Of the forty patients, thirty-six had a successful reduction. No premedication was required in twenty-nine patients who had a successful reduction, and the average time required for reduction in twenty patients was less than two minutes. Only four patients reported severe pain during the process of reduction. The method was not successful in four patients, two of whom had a displaced fracture of the greater tuberosity. CONCLUSIONS: The external rotation method for the reduction of an acute anterior dislocation of the shoulder is a safe and reliable method that can be performed relatively painlessly for both subcoracoid and subglenoid dislocations provided that a displaced fracture of the greater tuberosity is not present. PMID- 15523015 TI - In-hospital cost of total hip arthroplasty in Canada and the United States. AB - BACKGROUND: There is little information comparing the costs of specific surgical procedures performed in Canada and those done in the United States. The objective of this study was to compare the in-hospital costs associated with primary total hip arthroplasty performed in the two countries. METHODS: In-hospital costs of 1679 consecutive patients (940 Canadian and 739 American patients) who underwent total hip arthroplasty were extracted from three Canadian and three United States teaching hospitals between 1997 and 2001. Participating hospitals used the same cost accounting system to provide per-patient demographic, clinical, and cost data. Canadian dollar costs were converted to United States dollar costs with use of purchasing power parities. RESULTS: The baseline clinical characteristics of patients undergoing total hip arthroplasty in Canada and the United States were similar. The American patients were a mean of 4.6 years older than the Canadian patients (mean [and standard deviation], 67.8 +/- 12.4 years compared with 63.2 +/- 14.9 years). The median cost for the primary arthroplasty was $6080 (mean [and standard error of the mean], $6766 +/-$119) at the three Canadian hospitals and $12,846 (mean, $13,339 +/-$131) at the United States hospitals (p < 0.0001). The mean length of stay (and standard deviation) was 7.2 +/- 4.7 days for the Canadian patients and 4.2 +/- 2.0 days for the American patients. Implants at one hospital in the United States were found to be four times more costly than those in a Canadian hospital. CONCLUSIONS: Higher in-hospital costs were found for the American hospitals despite the fact that they had a significantly shorter patient length of stay compared with Canadian centers (p < 0.0001). Canadian hospitals should follow the lead of their counterparts in the United States and implement strategies to decrease the length of stay in the hospital, while institutions in the United States should revisit their ability to better manage the costs related to a primary total hip arthroplasty, particularly by controlling unit costs. PMID- 15523016 TI - Ununited diaphyseal forearm fractures with segmental defects: plate fixation and autogenous cancellous bone-grafting. AB - BACKGROUND: With current techniques of plate-and-screw fixation, diaphyseal nonunions of the radius and ulna are unusual. The few reports that have been published have discussed the use of structural corticocancellous bone grafts for the treatment of atrophic nonunions that are associated with osseous defects. We reviewed the rate of union and the functional results in association with the use of plate-and-screw fixation and autogenous cancellous (nonstructural) bone grafts. METHODS: Thirty-five patients with an atrophic ununited diaphyseal fracture of the forearm were treated with 3.5-mm plate-and-screw fixation and autogenous cancellous bone-grafting. A segmental osseous defect with an average size of 2.2 cm (range, 1 to 6 cm) was present in each patient. Twenty of the original fractures had been open. Eleven patients had had treatment of a deep infection before referral to us. The nonunion involved both forearm bones in eight patients, the radius alone in sixteen patients, and the ulna alone in eleven patients. RESULTS: The atrophic nonunion was associated with an open fracture in twenty patients, suboptimal fixation in twenty-two, a fracture dislocation of the forearm in nine, and infection in eleven. All fractures healed without additional intervention within six months. Two patients had a subsequent Darrach resection of the distal part of the ulna for the treatment of arthrosis of the distal radioulnar joint. After an average duration of follow-up of forty three months, the final arc of motion averaged 121 degrees in the forearm, 131 degrees at the elbow, and 137 degrees at the wrist, with an average grip strength of 83% compared with that of the contralateral limb. According to the system of Anderson and colleagues, five patients had an excellent result, eighteen had a satisfactory result, eleven had an unsatisfactory result (because of elbow stiffness related to associated elbow injuries in three and because of wrist stiffness in eight), and one had a poor result (because of malunion). CONCLUSIONS: When the soft-tissue envelope is compliant, has limited scar, and consists largely of healthy muscle with a good vascular supply, autogenous cancellous bone-grafting and stable internal plate fixation results in a high rate of union and improved upper limb function in patients with diaphyseal nonunion of the radius and/or ulna. PMID- 15523017 TI - Cost-effectiveness analysis of total ankle arthroplasty. AB - BACKGROUND: There is renewed interest in total ankle arthroplasty as an alternative to ankle fusion in the treatment of end-stage ankle arthritis. Despite a lack of long-term data on the clinical outcomes associated with these implants, the use of ankle arthroplasty is expanding. The purpose of this cost effectiveness analysis was to evaluate whether the currently available literature justifies the emerging use of total ankle arthroplasty. This study also identifies thresholds for the durability and function of ankle prostheses that, if met, would support more widespread dissemination of this new technology. METHODS: A decision model was created for the treatment of ankle arthritis. The literature was reviewed to identify possible outcomes and their probabilities following ankle fusion and ankle arthroplasty. Each outcome was weighted for quality of life with use of a utility factor, and effectiveness was expressed in units of quality-adjusted life years. Gross costs were estimated from Medicare charge and reimbursement data for the relevant codes. The effect of the uncertainty of estimates of costs and effectiveness was assessed with sensitivity analysis. RESULTS: The reference case of our model assumed a ten-year duration of survival of the prosthesis, resulting in an incremental cost-effectiveness ratio for ankle arthroplasty of $18,419 per quality-adjusted life year gained. This reflects a gain of 0.52 quality-adjusted life years at a cost of $9578 when ankle arthroplasty is chosen over fusion. This ratio compares favorably with the cost effectiveness of other medical and surgical interventions. Sensitivity analysis determined that the cost per quality-adjusted life year gained with ankle arthroplasty rises above $50,000 if the prosthesis is assumed to fail before seven years. Treatment options with ratios above $50,000 per quality-adjusted life year are commonly considered to have limited cost-effectiveness. This threshold is also crossed when the theoretical functional advantages of ankle arthroplasty are eliminated in sensitivity analysis. CONCLUSIONS: The currently available literature has not yet shown that total ankle arthroplasty predictably results in levels of durability and function that make it cost-effective at this time. However, the reference case of this analysis does demonstrate that total ankle arthroplasty has the potential to be a cost-effective alternative to ankle fusion. This reference case assumes that the theoretical functional advantages of ankle arthroplasty over ankle fusion will be borne out in future clinical studies. Performance of total ankle replacement will be better justified if these thresholds are met in published long-term clinical trials. PMID- 15523018 TI - A comparison of radiographic and scintigraphic techniques to assess aseptic loosening of the acetabular component in a total hip replacement. AB - BACKGROUND: The diagnosis of a loose total hip prosthesis is often established with use of radiographic and nuclear medicine techniques, but there is controversy about the relative utility of plain radiography, subtraction arthrography, nuclear arthrography, and bone scintigraphy. In this retrospective study, we evaluated the sensitivity, specificity, and interobserver reliability of these imaging modalities in patients suspected of having aseptic loosening of the acetabular component. METHODS: From 1994 to 1999, eighty-six consecutive patients with pain after a total hip arthroplasty were evaluated for possible loosening of the components. The imaging evaluation included plain radiography followed by a one-day protocol that included bone scintigraphy, subtraction arthrography, and nuclear arthrography. For this study, two experienced nuclear medicine physicians and two experienced radiologists, all of whom were blinded with respect to the clinical pretest data and the clinical outcome, retrospectively interpreted the diagnostic images. The sensitivity and the specificity of each imaging modality were established by comparing the findings obtained with each technique with those found at surgery or during the subsequent clinical course of the patient. Interobserver variability was determined with the intraclass correlation coefficient. RESULTS: Plain radiography had a sensitivity of 85% (95% confidence interval, 71 to 94) and a specificity of 85% (95% confidence interval, 66 to 96) in detecting aseptic loosening of the acetabular component, but it had only fair interobserver variability (intraclass correlation coefficient, 0.37). For subtraction arthrography, the sensitivity was 72% (95% confidence interval, 57 to 84), the specificity was 70% (95% confidence interval, 50 to 86), and there was good interobserver variability (intraclass correlation coefficient, 0.71). For nuclear arthrography, the sensitivity was 57% (95% confidence interval, 41 to 71), the specificity was 67% (95% confidence interval, 46 to 84), and there was fair interobserver variability (intraclass correlation coefficient, 0.24). For bone scintigraphy, the sensitivity was 83% (95% confidence interval, 69 to 92), the specificity was 67% (95% confidence interval, 46 to 84), and there was moderate interobserver variability (intraclass correlation coefficient, 0.43). CONCLUSIONS: Plain radiography had the highest diagnostic accuracy in the evaluation of aseptic loosening of the acetabular component. The diagnostic accuracy was increased when plain radiography was combined with bone scintigraphy or subtraction arthrography. However, we found considerable interobserver variability in image interpretation, even with experienced radiologists and nuclear medicine physicians. PMID- 15523019 TI - Patient-reported outcome and survivorship after Kinemax total knee arthroplasty. AB - BACKGROUND: We examined the long-term survivorship and patient-reported outcomes at a minimum of ten years following primary total knee arthroplasty. We hypothesized (1) that the survival rate would be at least 90% at ten years; (2) that age, gender, body-mass index, and primary diagnosis would not affect the survival rate; and (3) that the functional status of patients would be comparable with that of an age and gender-matched normal population. METHODS: A total of 407 patients (523 knees) who had had primary total knee arthroplasty between January 1988 and April 1991 were identified. The mean age of the patients at the time of surgery was sixty-nine years, and 68% of the patients were women. At ten years, 165 patients (211 knees) had died; seven of these 211 knees had been revised before the time of death. Of the remaining 242 patients, 208 (86%) completed a questionnaire, which included the Western Ontario and McMaster University Osteoarthritis Index (WOMAC), the Short Form-36 (SF-36), and questions regarding patient satisfaction and revision surgery. In the group of patients who participated in the study, ten patients (eleven knees) had had a revision before the review. RESULTS: A total of eighteen knees were revised. Twelve knees were revised because of aseptic failure and, of these, nine were revised because of polyethylene wear. The probability of survival at ten years was 96.1% with revision for any reason as the end point (and 97.2% when only aseptic failures were considered). Because of the small number of failures, we were unable to draw conclusions about associations between failure and age, gender, diagnosis, and body-mass index. The mean WOMAC scores (and standard deviation) at the time of the evaluation were 88 +/- 17 for pain and 79 +/- 20 for function. The SF-36 scores were similar to those for an age and gender-matched normal population, with only the physical functioning score being significantly lower (p < 0.001) and with the general health score being significantly higher (p < 0.001). Patients generally were very satisfied with all aspects of the outcome. CONCLUSIONS: At ten years, the survival of the prosthesis was excellent and the majority of patients were functionally independent, had very little knee pain, and were very satisfied with the result. The health benefits of this total knee arthroplasty were maintained after a minimum duration of follow-up of ten years. PMID- 15523020 TI - Histologic analysis of acetabular and proximal femoral bone in patients with osteonecrosis of the femoral head. AB - BACKGROUND: The purpose of this study was to investigate the hypothesis that osteonecrosis of the femoral head may also involve the acetabulum and the proximal part of the femur. METHODS: Twenty-five patients who underwent simultaneous bilateral total hip arthroplasty and thirty-eight patients who had a unilateral total hip arthroplasty for treatment of symptomatic osteonecrosis of the femoral head were included in the study. There were fifty-five men and eight women. The etiology of the osteonecrosis was idiopathic in forty patients (fifty six hips, 64%), associated with ethanol abuse in eighteen patients (twenty-five hips, 28%), and associated with steroid use in five patients (seven hips, 8%). The mean age of the patients at the time of arthroplasty was 47.1 years. We performed cancellous bone biopsies in the acetabulum, the proximal part of the femur, and the femoral head intraoperatively and then examined the specimens histologically. RESULTS: Of the eighty-one hips with idiopathic or ethanol associated osteonecrosis, seventy-six (94%) had normal or stage-1 bone in the acetabulum and the proximal part of the femur according to the system of Arlet and Ficat, and seventy-eight (97%) of the eighty-one had a grade of 0 or 1 according to the system of Humphreys et al. Of the seven hips with steroid associated osteonecrosis, four had normal or stage-1 bone and a grade of 0 or 1 and three had stage-2 or 3 disease and a grade of 2 or 3. Therefore, the vast majority of hips with idiopathic or ethanol-associated osteonecrosis had normal or nearly normal bone in the acetabulum and the proximal part of the femur. Three of the seven hips with steroid-associated osteonecrosis had abnormal bone in the proximal part of the femur only. CONCLUSIONS: One can expect to find normal or nearly normal cancellous bone in the acetabulum and the proximal part of the femur of patients with either idiopathic or ethanol-associated osteonecrosis of the hip. More extensive osteonecrosis may accompany steroid-associated osteonecrosis of the hip. PMID- 15523021 TI - Cementless Metasul metal-on-metal total hip arthroplasty in patients less than fifty years old. AB - BACKGROUND: Durable results of total hip arthroplasty have been difficult to achieve in young patients. We reviewed the intermediate-term clinical and radiographic results in a series of active, higher-demand patients who were less than fifty years old when they underwent cementless total hip arthroplasty with the use of the Metasul metal-on-metal articulation. METHODS: Seventy total hip arthroplasties were performed in sixty-two patients who were younger than fifty years of age (average age, thirty-seven years). Two patients (two hips) had had a resection arthroplasty because of deep infection less than five years postoperatively and were excluded. Sixty patients (sixty-eight hips) were available for complete clinical and radiographic analysis after a mean duration of follow-up of seven years. RESULTS: The mean preoperative Harris hip score of 49 points improved to 95 points at the time of final follow-up; fifty-six patients (93%) had an excellent result. No component was seen to be loose radiographically at the time of final follow-up. Only one focal area of pelvic osteolysis in one patient and two small focal areas of femoral osteolysis in another patient were identified. The hip with focal pelvic osteolysis underwent revision surgery with a liner change and bone-grafting of the osteolytic lesion around a stable component. CONCLUSIONS: At a mean of seven years after arthroplasties with a Metasul metal-on-metal articulation, there was a low rate of osteolysis and aseptic loosening in this group of young patients. However, additional follow-up is necessary to determine any possible long-term deleterious effects associated with this metal-on-metal articulation. PMID- 15523022 TI - Effect of gap size on gliding resistance after flexor tendon repair. AB - BACKGROUND: Gap formation is a common complication after flexor tendon repair and is associated with adhesion formation, tendon rupture, and decreased strength. The purpose of this study was to investigate the effect of gap formation on tendon gliding resistance after flexor tendon repair in a human cadaver model. METHODS: Twelve index, middle, and ring fingers from four adult human cadaveric hands were used. Gliding resistance versus excursion between the flexor digitorum profundus tendon and the A2 pulley was first measured in intact tendons. After full laceration, each tendon was repaired with the Pennington suture technique and the gliding resistance was measured again. Then, the repaired tendon (a 0-mm gap) was stretched to form a 1-mm gap, and gliding resistance was remeasured. A magnified video image was used to monitor gap size. This process was repeated to evaluate gap sizes of 2, 3, and 4 mm at the repair site. Peak gliding resistance was determined, and the peak gliding resistance was compared among the groups. RESULTS: No significant difference in peak gliding resistance was detected between repaired tendons without a gap and tendons with a 1-mm gap. Repaired tendons with a 2-mm gap could pass through the A2 pulley; however, peak gliding resistance was significantly higher than that for tendons with a 0 or a 1-mm gap (p < 0.05). When the gap reached > or =3 mm, all tendons caught at the A2 pulley edge, causing a dramatically increased peak gliding resistance. CONCLUSIONS: The presence of a 2-mm gap after flexor tendon repair significantly increased tendon peak gliding resistance (p < 0.05), while a gap of > or =3 mm further increased peak gliding resistance because of catching at the pulley edge. PMID- 15523023 TI - Shoulder arthroplasty for the treatment of inflammatory arthritis. AB - BACKGROUND: Prosthetic replacement of the glenohumeral joint can relieve pain and improve shoulder function for patients with end-stage inflammatory arthritis. The purpose of this study was to prospectively analyze the clinical, functional, and radiographic outcomes of shoulder reconstruction with hemiarthroplasty or total shoulder arthroplasty. METHODS: In this multicenter prospective study, clinical history, physical examination, and self-assessment tools including a visual analogue scale, the Simple Shoulder Test, and an activities questionnaire were used to measure comfort, quality of life, and function. Radiographic outcome was determined by assessing the severity of the disease, the adaptation of the prosthesis to the anatomy, the implant position and relationships, and the restoration of glenohumeral alignment. RESULTS: At the time of follow-up, at a minimum of twenty-four months (mean, thirty-nine months), the thirty-six shoulders treated with a hemiarthroplasty and the twenty-five treated with a total shoulder arthroplasty showed significant improvement (p < 0.0001) as demonstrated by the visual analogue scale and the Simple Shoulder Test as well as improvements in the components of the activities questionnaire. Active forward elevation was significantly better (p < 0.004) after the total shoulder arthroplasties than after the hemiarthroplasties. The presence of extremely severe disease did not affect the clinical outcome. Prosthetic adaptation to the anatomy and restoration of glenohumeral alignment resulted in significant improvement in certain motion parameters and were associated with one another (p < 0.001). Restoration of glenohumeral alignment resulted in significant improvements in overall quality of life (p = 0.038), use of the arm for work and play (p = 0.014), and range of motion (p = 0.0004) compared with those parameters when alignment had not been restored. Glenoid erosion occurred in four of the shoulders treated with hemiarthroplasty. Two of the glenoid components used in the total shoulder arthroplasties loosened. CONCLUSIONS: Patients with inflammatory arthritis treated with hemiarthroplasty or total shoulder arthroplasty can be expected to have improved comfort, range of motion, and function. Restoration of glenohumeral alignment appears to lead to even greater improvement in these clinical parameters. PMID- 15523024 TI - Systemic Vibrio infection presenting as necrotizing fasciitis and sepsis. A series of thirteen cases. AB - BACKGROUND: Vibrio species are an uncommon cause of necrotizing fasciitis and primary septicemia, which are likely to occur in patients with hepatic disease, diabetes mellitus, adrenal insufficiency, and immunocompromised conditions. These organisms are found in warm sea waters and are often present in raw oysters, shellfish, and other seafood. The purposes of the present report were to describe a series of patients who had this potentially lethal infection and to identify clinical features associated with a poor prognosis. METHODS: We retrospectively reviewed the records of thirteen patients (ten men and three women) who had necrotizing fasciitis and sepsis caused by Vibrio species. All patients had a history of contact with seawater or raw seafood. Eight patients had a hepatic disease such as hepatitis or cirrhosis of the liver, three had diabetes mellitus (without hepatic disease), and two had chronic renal or adrenal insufficiency (without hepatic disease). RESULTS: Twelve patients underwent fasciotomy or limb amputation. Five patients (38%) died within two to six days after admission, and eight patients survived. Patients with a systolic blood pressure of < or =90 mm Hg and leukopenia in the emergency room had a significantly higher mortality rate (p < 0.05). CONCLUSIONS: The diagnosis of Vibrio necrotizing fasciitis should be suspected when a patient has the appropriate clinical findings and a history of contact with seawater or raw seafood. The treatment should begin as early as possible, essentially when the patient has symptoms of sepsis. Although emergency fasciotomy or limb amputation did not reduce the mortality rate in this series, we consider such operations to be an important aspect of treatment. PMID- 15523025 TI - Vertebral osteonecrosis associated with pancreatitis in a woman with pancreas divisum. A case report. PMID- 15523026 TI - Pseudarthrosis of a thirty-nine-year-old dens fracture causing myelopathy. A case report. PMID- 15523027 TI - Type-III dens fracture with distraction: an unstable injury. A report of three cases. PMID- 15523029 TI - Intraosseous leiomyoma of the ulna. A case report. PMID- 15523028 TI - Total knee arthroplasty using recombinant factor VII in hemophilia-A patients with inhibitors. A report of three cases. PMID- 15523030 TI - Hydroxyapatite-coated prostheses in total hip and knee arthroplasty. AB - Hydroxyapatite-coated implants have demonstrated extensive bone apposition in animal models. The osseous interface develops even in the presence of gaps of 1 mm and relative motion of up to 500 mum. Development of implant-bone interfacial strength is due to the biological effects of released calcium and phosphate ions, although surface roughness leads to increased interface strength in the absence of interface gaps. The clinical results at fifteen years after total hip replacements have demonstrated that hydroxyapatite-coated femoral stems perform as well as, and possibly better than, other types of cementless devices, with the added benefit of providing a seal against wear debris. Hydroxyapatite-coated acetabular components must have a mechanical interlock with bone in order to take advantage of the coating effects. Clinical analyses of these types of designs at seven years have indicated good survivorship. The performance of a hydroxyapatite coated implant depends on coating properties (thickness, porosity, hydroxyapatite content, and crystallinity), implant roughness, and overall design. The most reliable predictor of the performance of a device is success in long-term clinical studies. PMID- 15523031 TI - Leading beyond the shadow-line. PMID- 15523032 TI - The future of specialization within orthopaedics. PMID- 15523033 TI - Fluoroscopically guided low-volume peritendinous corticosteroid injection for Achilles tendinopathy. A safety study. PMID- 15523034 TI - Therapeutic effects of hyaluronic acid on osteoarthritis of the knee. PMID- 15523035 TI - Open fractures of the calcaneus: soft-tissue injury determines outcome. PMID- 15523036 TI - Load-sharing at the wrist following radial head replacement with a metal implant. A cadaveric study. PMID- 15523037 TI - Ligament reconstruction versus trapezial resection alone for thumb carpometacarpal osteoarthritis. PMID- 15523038 TI - Bosworth fracture-dislocation and resultant compartment syndrome. PMID- 15523039 TI - Total knee arthroplasty following proximal tibial osteotomy. PMID- 15523040 TI - Computer-assisted knee arthroplasty is better than a conventional jig-based technique in terms of component alignment. PMID- 15523041 TI - Arthroscopic repair is as effective as open repair for isolated Bankart lesions of the shoulder and may improve range of motion. PMID- 15523042 TI - Olpadronate reduced fractures in children with osteogenesis imperfecta. PMID- 15523043 TI - What's new in orthopaedic rehabilitation. PMID- 15523044 TI - The moving frontier in nitric oxide-dependent signaling. AB - New discoveries are expanding our view of the role of nitric oxide (NO) in mammalian physiology by revealing new types, amounts, and fates of molecules modified in vivo by NO and its derivatives, as well as the profound augmentation of some of NO's actions at physiologic levels of O(2). Investigators have identified a new form of endogenous reversible N-nitrosation reactions in vivo, that of proteins; a new class of endogenously nitrated bioactive molecules in vivo, nitro-fatty acids; and the ability of NO-dependent posttranslational modifications to control the half-life and destination of key regulatory proteins. PMID- 15523045 TI - Pharmacokinetics of tacrolimus and mycophenolic acid are altered, but recover at different times during hepatic regeneration in rats. AB - Hepatic regeneration is very critical to the success of living donor liver transplantation, which allows a reduced size liver to grow in size to accommodate the requirements of both the donor and the recipient. The objectives of this study were to evaluate 1) the hepatic metabolism of the two immunosuppressive drugs, tacrolimus and mycophenolic acid (MPA), and 2) the pharmacokinetics of tacrolimus and mycophenolic acid at various time points after initiation of hepatic regeneration by partial hepatectomy in rats. The hepatic intrinsic clearance of tacrolimus was decreased to 70% and 51% of the control level at the 24th h and the 6th day, respectively, but returned to normal level by day 14. The total body clearance of tacrolimus was reduced transiently but recovered completely by day 18. The hepatic intrinsic clearance of MPA was decreased to 52% and 51% of that in control rats at the 24th h and the 6th day, respectively, but recovered to normal level by day 14. The total body clearance of MPA was reduced at the 24th h but recovered by day 6. The magnitude of reduction in the clearance of tacrolimus and MPA was much smaller than what was predicted from in vitro data. The elimination clearance of MPA glucuronide was also impaired during hepatic regeneration but recovered to normal level with time. In conclusion, the pharmacokinetics of tacrolimus and mycophenolic acid were altered during hepatic regeneration but recovered completely at different rates over time. Caution must be exercised in extrapolating in vitro data to in vivo conditions during hepatic regeneration. PMID- 15523046 TI - Bioactivation of the nontricyclic antidepressant nefazodone to a reactive quinone imine species in human liver microsomes and recombinant cytochrome P450 3A4. AB - The therapeutic benefits of the antidepressant nefazodone have been hampered by several cases of acute hepatotoxicity/liver failure. Although the mechanism of hepatotoxicity remains unknown, it is possible that reactive metabolites of nefazodone play a causative role. Studies were initiated to determine whether nefazodone undergoes bioactivation in human liver microsomes to electrophilic intermediates. Following incubation of nefazodone with microsomes or recombinant P4503A4 in the presence of sulfydryl nucleophiles, conjugates derived from the addition of thiol to a monohydroxylated nefazodone metabolite were observed. Product ion spectra suggested that hydroxylation and sulfydryl conjugation occurred on the 3-chlorophenylpiperazine-ring, consistent with a bioactivation pathway involving initial formation of p-hydroxynefazodone, followed by its two electron oxidation to the reactive quinone-imine intermediate. The formation of novel N-dearylated nefazodone metabolites was also discernible in these incubations, and 2-chloro-1,4-benzoquinone, a by-product of N-dearylation, was trapped with glutathione to afford the corresponding hydroquinone-sulfydryl adduct. Nefazodone also displayed NADPH-, time-, and concentration-dependent inactivation of P4503A4 activity, suggesting that reactive metabolites derived from nefazodone bioactivation are capable of covalently modifying P4503A4. A causative role for 2-chloro-1,4-benzoquinone and/or the quinone-imine intermediate(s) in nefazodone hepatotoxicity is speculated. Although the antianxiety agent buspirone, which contains a pyrimidine ring in place of the 3 chlorophenyl-ring, also generated p-hydroxybuspirone in liver microsomes, no sulfydryl conjugates of this metabolite were observed. This finding is consistent with the proposal that two-electron oxidation of p-hydroxybuspirone to the corresponding quinone-imine is less favorable due to differences in the protonation state at physiological pH and due to weaker resonance stabilization of the oxidation products as predicted from ab initio measurements. PMID- 15523047 TI - Formation and protein binding of the acyl glucuronide of a leukotriene B4 antagonist (SB-209247): relation to species differences in hepatotoxicity. AB - SB-209247 [(E)-3-[6-[[(2,6-dichlorophenyl)-thio]methyl]-3-(2-phenylethoxy)-2 pyridinyl]-2-propenoic acid], an anti-inflammatory leukotriene B4 receptor antagonist, was associated in beagle dogs but not male rats with an inflammatory hepatopathy. It also produced a concentration-dependent (10-1000 microM) but equal leakage of enzymes from dog and rat precision-cut liver slices. The hepatic metabolism of SB-209247 was investigated with reference to the formation of reactive acyl glucuronides. [14C]SB-209247 (100 micromol/kg) administered i.v. to anesthetized male rats was eliminated by biliary excretion of the acyl glucuronides of the drug and its sulfoxide. After 5 h, 1.03 +/- 0.14% (mean +/- S.E.M., n = 4) of the dose was bound irreversibly to liver tissue. The sulfoxide glucuronide underwent pH-dependent rearrangement in bile more rapidly than did the SB-209247 conjugate. [14C]SB-209247 was metabolized by sulfoxidation and glucuronidation in rat and dog hepatocytes, and approximately 1 to 2% of [14C]SB 209247 (100 microM) became irreversibly bound to cellular material. [14C]SB 209247 sulfoxide and glucuronide were the only metabolites produced by dog, rat, and human liver microsomes in the presence of NADPH and UDP-glucuronic acid (UDPGA), respectively. V(max) values for [14C]SB-209247 glucuronidation by dog, rat, and human microsomes were 2.6 +/- 0.1, 1.2 +/- 0.1, and 0.4 +/- 0.0 nmol/min/mg protein, respectively. Hepatic microsomes from all three species catalyzed UDPGA-dependent but not NADPH-dependent irreversible binding of [14C]SB 209247 (100-250 microM) to microsomal protein. Although a reactive acyl glucuronide was formed by microsomes from every species, the binding did not differ between species. Therefore, neither the acute cellular injury nor glucuronidation-driven irreversible protein binding in vitro is predictive of the drug-induced hepatopathy. PMID- 15523048 TI - Impact of transcription factor profile and chromatin conformation on human hepatocyte CYP3A gene expression. AB - Recent data have made it increasingly clear that the gene expression profile of a cell system, and its alteration in response to external stimuli, is highly dependent on both the higher order chromatin structure of the genome and the interaction of gene products in interpreting stimuli. To further explore this phenomenon, we have examined the role of both of these factors in controlling xenobiotic-mediated gene expression changes in primary and transformed human hepatocytes (HuH7). Using quantitative polymerase chain reaction, expression levels of several transcription factors implicated in the liver-specific regulation of the CYP3A gene family were examined in human adult and fetal liver RNA samples. These expression profiles were then compared with those obtained from both primary and transformed human hepatocytes, showing that, in general, cultured cells exhibit a distinct profile compared with either the fetal or adult samples. Transcriptome profiles before and after exposure to the CYP3A transcriptional activators rifampicin, dexamethasone, pregnane-16alpha carbonitrile, and phenobarbital were subsequently examined. Whereas exposure to these compounds elicited a dose-dependent increase in CYP3A transcription in primary hepatocytes, no alteration in expression levels was observed for the hepatoma cell line HuH7. Alteration in the expression levels of pregnane X receptor and chicken ovalbumin upstream promoter transcription factor I, and the disruption of higher order chromatin within HuH7 cells altered CYP3A expression and/or activation by xenobiotics toward that observed in primary hepatocytes. These data provide potential roles for these two processes in regulating CYP3A expression in vivo. PMID- 15523049 TI - Mapping of multidrug resistance gene 1 and multidrug resistance-associated protein isoform 1 to 5 mRNA expression along the human intestinal tract. AB - Efflux transporters such as P-glycoprotein and multidrug resistance-associated proteins (MRPs) in the intestinal wall restrict intestinal drug transport. To overcome this limitation for enteral drug absorption, galenical targeting approaches have been proposed for site-specific luminal drug release in segments of the gut, where expression of the respective absorption-limiting transporter is minimal. Therefore, expression of multidrug resistance gene 1 (MDR1) and MRP1-5 was systematically investigated in 10 healthy subjects. Biopsies were taken from different segments of the gastrointestinal tract (from duodenum and terminal ileum, as well as ascending, transverse, descending, and sigmoid colon). Gene expression was investigated by quantitative real-time PCR (TaqMan). MRP3 appeared to be the most abundantly expressed transporter in investigated parts of the human intestine, except for the terminal ileum, where MDR1 showed the highest expression. The ranking of transporter gene expression in the duodenum was MRP3 >> MDR1 > MRP2 > MRP5 > MRP4 > MRP1. In the terminal ileum, the ranking order was as follows: MDR1 > MRP3 >> MRP1 approximately MRP5 approximately MRP4 > MRP2. In all segments of the colon (ascending, transverse, descending, and sigmoid colon), the transporter gene expression showed the following order: MRP3 >> MDR1 > MRP4 approximately MRP5 > MRP1 >> MRP2. We have shown, for the first time, systematic site-specific expression of MDR1 and MRP mRNA along the gastrointestinal tract in humans. All transporters showed alterations in their expression levels from the duodenum to sigmoid colon. The most pronounced changes were observed for MRP2, with high levels in the small intestine and hardly any expression in colonic segments. This knowledge may be useful to develop new targeting strategies for enteral drug delivery. PMID- 15523050 TI - C. difficile: by the numbers. PMID- 15523051 TI - Receptor-independent activation of GABAergic neurotransmission and receptor dependent nontranscriptional activation of phosphatidylinositol 3-kinase/protein kinase Akt pathway in short-term cardiovascular actions of dexamethasone at the nucleus tractus solitarii of the rat. AB - Whereas glucocorticoids are important blood pressure regulators via an action on peripheral circulation, their roles in central cardiovascular regulation are less known. This study evaluated the short-term cardiovascular effect of glucocorticoid in the nucleus tractus solitarii (NTS) and delineated the underlying molecular mechanisms. In Sprague-Dawley rats maintained under propofol anesthesia, microinjection bilaterally into the NTS of a synthetic glucocorticoid, dexamethasone (Dex; 12.5, 25, 50, or 100 pmol), elicited hypertensive and tachycardiac responses. The initial cardiovascular responses, which lasted 15 to 30 min, were blunted by coadministration of a selective GABA(A) or GABA(B) receptor antagonist, bicuculline (15 pmol) or 2-hydroxy saclofen (150 pmol). The delayed responses, which endured at least 90 min and entailed maintained hypertension and tachycardia, were reversed by selective glucocorticoid type II receptor (GR) antagonist mifepristone (100 or 200 pmol), phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 [2-(4-morpholinyl)-8 phenyl-4H-1-benzopyran-4-one] (20 nmol), or nitric-oxide synthase inhibitor N(G) monomethyl-l-arginine acetate (5 nmol), but not by the RNA synthesis inhibitor actinomycin D (20 nmol). Moreover, Dex induced an association of GR with the regulatory subunit of PI3K, p85alpha, in a ligand-dependent manner and promoted serine/threonine kinase Akt phosphorylation that was blocked by coadministration of mifepristone or LY294002. These cardiovascular and molecular responses occurred when translocation of activated GR into the nucleus was minimal. Our results indicate that Dex acts on the NTS to elicit hypertension and tachycardia via both a GR-independent interaction with GABA(A) and GABA(B) receptors and a GR dependent but nontranscriptional mechanism that involves activation of PI3K/Akt pathway. PMID- 15523053 TI - Helix I of beta-arrestin is involved in postendocytic trafficking but is not required for membrane translocation, receptor binding, and internalization. AB - beta-Arrestins bind to phosphorylated, seven-transmembrane-spanning, G protein coupled receptors (GPCRs), including the type 1 angiotensin II receptor (AT(1)R), to promote receptor desensitization and internalization. The AT(1) R is a class B GPCR that recruits both beta-arrestin1 and beta-arrestin2, forming stable complexes that cotraffic to deep-core endocytic vesicles. beta-Arrestins contain one amphipathic and potentially amphitropic (membrane-targeting) alpha-helix (helix I) that may promote translocation to the membrane or influence receptor internalization or trafficking. Here, we investigated the trafficking and function of beta-arrestin1 and beta-arrestin2 mutants bearing substitutions in both the hydrophobic and positively charged faces of helix I. The level of expression of these mutants and their cytoplasmic localization (in the absence of receptor activation) was similar to wild-type beta-arrestins. After angiotensin II stimulation, both wild-type and beta-arrestin mutants translocated to the cell membrane, although recruitment was weaker for mutants of the hydrophobic face of helix I. For all beta-arrestin mutants, the formation of deep-core vesicles was less observed compared with wild-type beta-arrestins. Furthermore, helix I conjugated to green fluorescent protein is not membrane-localized, suggesting that helix I, in isolation, is not amphitropic. Bioluminescence resonance energy transfer analysis revealed that both wild-type and beta-arrestin mutants retained a capacity to interact with the AT(1)R, although the interaction with the mutants was less stable. Finally, wild-type and mutant beta-arrestins fully supported receptor internalization in human embryonic kidney cells and mouse embryonic fibroblasts deficient in beta-arrestin1 and -2. Thus, helix I is implicated in postmembrane trafficking but is not strongly amphitropic. PMID- 15523052 TI - Steroidogenic factor-1 interacts with cAMP response element-binding protein to mediate cAMP stimulation of CYP1B1 via a far upstream enhancer. AB - CYP1B1 activates polycyclic aromatic hydrocarbon carcinogens in cAMP-regulated tissues such as the adrenal, ovary, and testis. A 27-fold cAMP stimulation of the CYP1B1-luciferase reporter in Y-1 adrenal cells depends entirely on a far upstream enhancer region (FUER; -5298 to -5110). Cooperative participation of multiple steroidogenic factor 1 (SF-1) elements with the downstream cAMP response element (CRE) in FUER is essential for both basal and cAMP-stimulated activities of FUER. Basal and induced activities were similarly lowered by DAX-1, an SF-1 suppressor, and raised by steroid receptor coactivator 1, an SF-1 coactivator. cAMP response element-binding protein (CREB)-binding protein (CBP) that interacts preferentially with the phosphorylated-CREB increased the cAMP-induced FUER. 10T1/2 cells and human embryonic kidney (HEK)293 cells do not express SF-1. Introduction of exogenous SF-1 generated cAMP stimulation of the FUER in 10T1/2 fibroblasts. The same transfection only increased basal activity of FUER in HEK293 cells, despite presence of active CREB in cells. HEK293 cells therefore remain deficient in additional factor(s) critical to the cAMP stimulation of CYP1B1. Mutations of the protein kinase A (PKA) and the mitogen-activated protein kinase phosphorylation sites (Ser-430 and Ser-203) on SF-1 had no effect on the SF-1-dependent FUER stimulation in Y-1 and 10T1/2 cells. This contrasts with loss of activity with mutation of CREB at PKA phosphorylation site (Ser-133). SF-1 phosphorylation at these sites is therefore not essential for the cAMP stimulation and the cooperation with CREB. cAMP-enhanced activation protein 1 (AP 1) and stimulatory protein 1 (Sp1) complexes in the proximal promoter region contributed substantially to both basal and cAMP-stimulated FUER activity. Chromatin immunoprecipitation from primary rat adrenal cells demonstrated cAMP stimulation of histone acetylation proximal to, respectively, the FUER and AP-1 sites of CYP1B1. PMID- 15523054 TI - PDZK1 directly regulates the function of organic cation/carnitine transporter OCTN2. AB - Urinary excretion of cationic xenobiotics is believed to be mediated by organic cation transporter (OCT and OCTN) families expressed on both basolateral and brush-border membranes of renal tubules, although the molecular mechanisms for targeting of these transporters to each membrane are poorly understood. Here, to examine the regulatory mechanisms for cell-surface expression and function of these transporters, we evaluated the interaction of these transporters with several PDZ proteins. A pull-down study using recombinant C-terminal proteins of OCTs and OCTNs identified a specific interaction of apical transporters OCTN1 and OCTN2, but not basolateral transporters OCT1 and OCT2, with PDZK1, intestinal and kidney-enriched PDZ protein, and Na+/H+ exchanger regulatory factor 2 (also called E3KARP, SIP-1, or TKA-1). Both yeast two-hybrid and pull-down studies suggested a requirement of the last four amino acids in OCTN1 and OCTN2 for the interaction. The interaction of PDZK1 with the C terminus of OCTN2 was also confirmed in a pull-down study using kidney brush-border membrane vesicles. Immunohistochemical analysis revealed that both PDZK1 and OCTN2 are colocalized in brush-border membranes of the kidney. Finally, double transfection of OCTN2 with PDZK1 stimulated the uptake by OCTN2 of its endogenous substrate carnitine, and this increase could be accounted for by the 6-fold increase in transport capacity. Such an increase was not observed for OCTN2 with deletion of the last four amino acids. Biotinylation study of surface proteins revealed minimal effect of PDZK1 on cell-surface expression of OCTN2. The present findings are the first to identify PDZK1 as a functional regulator of OCTN2 through direct interaction with the C terminus. PMID- 15523055 TI - 2004 Lasker Awards honors announced. PMID- 15523058 TI - Tool helps cancer scientists mine genes. PMID- 15523063 TI - Trauma and PTSD Symptoms in Rwanda. PMID- 15523064 TI - MMR vaccination and febrile seizures. PMID- 15523065 TI - Androgen suppression plus radiation therapy for prostate cancer. PMID- 15523066 TI - Androgen suppression plus radiation therapy for prostate cancer. PMID- 15523067 TI - Fetal loss associated with excess thyroid hormone exposure. PMID- 15523068 TI - History of emerging vascular disease risk factors. PMID- 15523069 TI - Annual revaccination against influenza and mortality risk in community-dwelling elderly persons. AB - CONTEXT: Although large-scale observational studies have demonstrated the effectiveness of influenza vaccination, no large studies have systematically addressed the clinical benefit of annual revaccinations. OBJECTIVE: To investigate the effect of annual influenza revaccination on mortality in community-dwelling elderly persons. DESIGN, SETTING, AND PARTICIPANTS: A population-based cohort study using the computerized Integrated Primary Care Information (IPCI) database in the Netherlands including community-dwelling individuals aged 65 years or older from 1996 through 2002. For each year, we computed the individual cumulative exposure to influenza vaccination since study start. MAIN OUTCOME MEASURE: Association between the number of consecutive influenza vaccinations and all-cause mortality vs no vaccination after adjusting for age, sex, chronic respiratory and cardiovascular disease, hypertension, diabetes mellitus, renal failure, and cancer. RESULTS: The study population included 26,071 individuals, of whom 3485 died during follow-up. Overall, a first vaccination was associated with a nonsignificant annual reduction of mortality risk of 10% (hazard ratio [HR], 0.90; 95% confidence interval [CI], 0.78-1.03) while revaccination was associated with a reduced mortality risk of 24% (HR, 0.76; 95% CI, 0.70-0.83). Compared with a first vaccination, revaccination was associated with a reduced annual mortality risk of 15% (HR, 0.85; 95% CI, 0.75 0.96). During the epidemic periods this reduction was 28% (HR, 0.72; 95% CI, 0.53 0.96). Similar estimates were obtained for persons with and without chronic comorbidity and those aged 70 years or older at baseline. Overall, influenza vaccination is estimated to prevent 1 death for every 302 vaccinees at a vaccination coverage that varied between 64% and 74%. CONCLUSION: Annual influenza vaccination is associated with a reduction in all-cause mortality risk in a population of community-dwelling elderly persons, particularly in older individuals. PMID- 15523070 TI - Utilization of early invasive management strategies for high-risk patients with non-ST-segment elevation acute coronary syndromes: results from the CRUSADE Quality Improvement Initiative. AB - CONTEXT: The American College of Cardiology/American Heart Association (ACC/AHA) guidelines for the management of non-ST-segment elevation acute coronary syndromes (NSTE ACS) recommend early invasive management for high-risk patients, given the benefits with this approach demonstrated in randomized clinical trials. OBJECTIVES: To determine the use and predictors of early invasive management strategies (cardiac catheterization <48 hours following presentation) in high risk patients with NSTE ACS and to examine the association of early invasive management with mortality. DESIGN, SETTING, AND PATIENTS: The CRUSADE (Can Rapid Risk Stratification of Unstable Angina Patients Suppress Adverse Outcomes With Early Implementation of the ACC/AHA Guidelines) Quality Improvement Initiative evaluated care patterns and outcomes for 17,926 high-risk NSTE ACS patients (positive cardiac markers and/or ischemic electrocardiographic changes) based on ACC/AHA guidelines recommendations at 248 US hospitals with catheterization and revascularization facilities between March 2000 and September 2002. MAIN OUTCOME MEASURES: Use of early invasive management within 48 hours of presentation, predictors of early invasive management, and in-hospital mortality. Results Of the 17,926 patients analyzed, 8037 (44.8%) underwent early cardiac catheterization less than 48 hours following presentation. Predictors of early invasive management included cardiology care, younger age, lack of prior or current congestive heart failure, lack of renal insufficiency, ischemic electrocardiographic changes, positive cardiac markers, white race, and male sex. Patients treated with early invasive management were more likely to be treated with medications and interventions recommended by the ACC/AHA guidelines and had a lower risk of in-hospital mortality after adjusting for differences in clinical characteristics and after comparing propensity-matched pairs (2.5% vs 3.7%, P<.001). Conclusions An early invasive management strategy is not utilized in the majority of high-risk patients with NSTE ACS. This strategy appears to be reserved for patients without significant comorbidities and those cared for by cardiologists and is associated with a lower risk of in-hospital mortality. PMID- 15523071 TI - Differential genetic effects of ESR1 gene polymorphisms on osteoporosis outcomes. AB - CONTEXT: Both bone mineral density (BMD) and fracture risk have a strong genetic component. Estrogen receptor alpha (ESR1) is a candidate gene for osteoporosis, but previous studies of ESR1 polymorphisms in this field were hampered by small sample size, lack of standardization, and inconclusive results. OBJECTIVE: To generate large-scale evidence on whether 3 common ESR1 polymorphisms (intron 1 polymorphisms XbaI [dbSNP: rs9340799] and PvuII [dbSNP: rs2234693] and promoter TA repeats microsatellite) and haplotypes thereof are associated with BMD and fractures. DESIGN AND SETTING: Meta-analysis of individual-level data involving standardized genotyping of 18 917 individuals in 8 European centers. MAIN OUTCOME MEASURES: BMD of femoral neck and lumbar spine; all fractures and vertebral fractures by genotype. RESULTS: No between-center heterogeneity was observed for any outcome in any genetic contrast. None of the 3 polymorphisms or haplotypes had any statistically significant effect on BMD in adjusted or unadjusted analyses, and estimated differences between genetic contrasts were 0.01 g/cm2 or less. Conversely, we found significant reductions in fracture risk. In women homozygous for the absence of an XbaI recognition site, the adjusted odds of all fractures were reduced by 19% (odds ratio, 0.81 [95% CI, 0.71-0.93]; P = .002) and vertebral fractures by 35% (odds ratio, 0.65 [95% CI, 0.49-0.87]; P = .003). Effects on fractures were independent of BMD and unaltered in adjusted analyses. No significant effects on fracture risk were seen for PvuII and TA repeats. CONCLUSIONS: ESR1 is a susceptibility gene for fractures, and XbaI determines fracture risk by mechanisms independent of BMD. Our study demonstrates the value of adequately powered studies with standardized genotyping and clinical outcomes in defining effects of common genetic variants on complex diseases. PMID- 15523072 TI - Hospitalization, restricted activity, and the development of disability among older persons. AB - CONTEXT: Preventing the development of disability in activities of daily living is an important goal in older adults, yet relatively little is known about the disabling process. OBJECTIVES: To evaluate the relationship between 2 types of intervening events (hospitalization and restricted activity) and the development of disability and to determine whether this relationship is modified by the presence of physical frailty. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort study, conducted in the general community in greater New Haven, Conn, from March 1998 to March 2003, of 754 persons aged 70 years or older, who were not disabled (ie, required no personal assistance) in 4 essential activities of daily living: bathing, dressing, walking inside the house, and transferring from a chair. Participants were categorized into 2 groups according to the presence of physical frailty (defined on the basis of slow gait speed) and were followed up with monthly telephone interviews for up to 5 years to ascertain exposure to intervening events and determine the occurrence of disability. MAIN OUTCOME MEASURE: Disability, defined as the need for personal assistance in bathing, dressing, walking inside the house, or transferring from a chair. RESULTS: During the 5-year follow-up period, disability developed among 417 (55.3%) participants, 372 (49.3%) were hospitalized and 600 (79.6%) had at least 1 episode of restricted activity. The multivariable hazard ratios for the development of disability were 61.8 (95% confidence interval [CI], 49.0-78.0) within a month of hospitalization and 5.54 (95% CI, 4.27-7.19) within a month of restricted activity. Strong associations were observed for participants who were physically frail and those who were not physically frail. Hospital admissions for falls were most likely to lead to disability. Intervening events occurring more than a month prior to disability onset were not associated with the development of disability. The population-attributable fractions associated with new exposure to hospitalization and restricted activity, respectively, were 0.48 and 0.19; 0.40 and 0.20, respectively, for frail participants and 0.61 and 0.16, respectively, for nonfrail participants. CONCLUSIONS: Illnesses and injuries leading to either hospitalization or restricted activity represent important sources of disability for older persons living in the community, regardless of the presence of physical frailty. These intervening events may be suitable targets for the prevention of disability. PMID- 15523073 TI - Is there a doctor in the house? . . . Or the Senate? Physicians in US Congress, 1960-2004. AB - CONTEXT: The legislative and fiscal influences of Congress, as well as the continuing overall growth in health care spending as a portion of the gross domestic product, make congressional representation by physicians important because physicians have unique expertise in the impact of legislation on patient care and medical practice. OBJECTIVES: To describe physician representation in the US Congress between 1960 and 2004 and relate the results to past representation of physicians in Congress. DESIGN AND SETTING: A retrospective observational study of members of the US Congress from all 50 states and all represented territories, who served from January 1960 to April 2004 (including 108th Congress), using data available in public access databases and congressional biographical records. MAIN OUTCOME MEASURES: Physician representation in Congress, including occupation before taking office, state/territory of representation, sex, party affiliation, and time served. RESULTS: During the past 44 years, 25 (1.1%) of 2196 members of Congress were physicians. Physicians in Congress were more likely to be members of the Republican Party (60% vs 45.1% of all members, P = .007) and were similar to other members of Congress in mean years of service (9.2 years for physicians vs 12.3 years for all members, P = .09) and in sex distribution (4.0% female physicians vs 6.8% all female members, P = .57). Physicians in Congress represented 17 states, the Virgin Islands, and Puerto Rico. CONCLUSIONS: Physician representation in Congress is low and is in stark contrast with physician roles during the first century of the United States. However, the 8 physicians currently serving in Congress may be indicative of a shift toward more direct influence of physicians in national politics. PMID- 15523074 TI - Trends in the risks and benefits to patients with cancer participating in phase 1 clinical trials. AB - CONTEXT: In the past, cancer patients entering phase 1 studies confronted the prospects of high risk and unlikely benefit. Over the last decade, cancer drugs under development have become more targeted, and the clinical research environment has become more scrutinized. The impact of these changes on the risks and benefits to patients who participate in phase 1 cancer trials is unknown. OBJECTIVE: To determine trends in the rates of treatment-related (toxic) death, objective response, and serious toxicity and to identify factors associated with these outcomes. DATA SOURCES: We searched abstracts and journal articles reporting the results of phase 1 cancer treatment trials originally submitted to annual meetings of the American Society of Clinical Oncology (ASCO) from 1991 through 2002. STUDY SELECTION: We focused on published single-agent trials that enrolled patients with advanced solid tumors and excluded studies testing agents already approved by the US Food and Drug Administration at the time of the ASCO presentation. DATA EXTRACTION: Multiple observers independently extracted information on trial design, location, sponsorship, types of tumors treated, drug class, route of administration, and clinical outcomes. DATA SYNTHESIS: The overall toxic death rate for 213 studies (involving 6474 cancer patients) published in peer-reviewed journals was 0.54%, while the overall objective response rate was 3.8%. Toxic death rates decreased over the study period, from 1.1% over the first 4 years of the study (1991-1994) to 0.06% over the most recent 4-year period (1999-2002) (P<.01). Response rates also decreased but by proportionally much less. After adjusting for characteristics of the experimental trials and the investigational agents, the odds of a patient dying from an experimental treatment while participating in a trial submitted during the most recent 4-year period were less than one tenth those of a patient participating in a trial submitted during the first 4-year period (odds ratio, 0.09; 95% confidence interval, 0.01-0.67; P = .009). In comparison, the adjusted odds of a patient experiencing an objective response over the same time periods decreased by approximately half (odds ratio, 0.46; 95% confidence interval, 0.32-0.66; P<.001). CONCLUSIONS: The level of risk experienced by cancer patients who participate in phase 1 treatment trials appears to have improved over the 12-year period from 1991 through 2002. Because toxic death rates have decreased more quickly than have objective response rates, the ratio of risk to benefit may have also improved. These changes relate in part to the targeted and less-toxic nature of newer cancer drugs and are coincident with the increased attention that has been paid to the safety of clinical research over the time period we analyzed. PMID- 15523075 TI - Caring for the child with cancer at the close of life: "there are people who make it, and I'm hoping I'm one of them". AB - Approximately 25% of children with cancer die of their disease. Early in the course of a patient's illness, it is often impossible to determine whether the disease will be cured with cancer-directed treatment. When potentially curative therapy is no longer an option, the patient, family, and oncology team face enormous medical, psychological, and spiritual challenges. Optimal palliative care requires willingness on the part of the physician and caregiver team to engage the patient and family in discussions of their hopes and fears and to provide solace and support for emotional and physical pain. Using the comments of a child in the terminal phase of acute leukemia, his mother, and his physician, we describe opportunities and important lessons often revealed only when families and their caregivers face the end of a child's life. A broad-minded assessment of the patient's and family's physical, emotional, and spiritual needs and clarification of realistic goals and hopes not only improves the clinical care that the patient receives but also contributes to the sense of satisfaction and meaning that the physician can gain from the experience of caring for children at the end of life. PMID- 15523076 TI - Risks and benefits of phase 1 clinical trials evaluating new anticancer agents: a case for more innovation. PMID- 15523077 TI - JAMA patient page. Influenza. PMID- 15523078 TI - Could exposure assessment problems give us wrong answers to nutrition and cancer questions? PMID- 15523079 TI - Nanotechnology grows into billion-dollar enterprise. PMID- 15523080 TI - Canadian province seeks control of its genes. PMID- 15523081 TI - Stat bite: Estimated deaths attributable to variations in health care. PMID- 15523082 TI - Researchers optimistic about targeted drugs for pancreatic cancer. PMID- 15523083 TI - Screening methods may offer early diagnosis of pancreatic cancer. PMID- 15523084 TI - Good drug, bad luck: business, regulatory issues can create obstacles for drug development. PMID- 15523085 TI - Last decade brought new guidelines, detection methods for colon cancer screening. PMID- 15523086 TI - Fruit and vegetable intake and risk of major chronic disease. AB - BACKGROUND: Studies of fruit and vegetable consumption in relation to overall health are limited. We evaluated the relationship between fruit and vegetable intake and the incidence of cardiovascular disease and cancer and of deaths from other causes in two prospective cohorts. METHODS: A total of 71 910 female participants in the Nurses' Health study and 37,725 male participants in the Health Professionals' Follow-up Study who were free of major chronic disease completed baseline semiquantitative food-frequency questionnaires in 1984 and 1986, respectively. Dietary information was updated in 1986, 1990, and 1994 for women and in 1990 and 1994 for men. Participants were followed up for incidence of cardiovascular disease, cancer, or death through May 1998 (women) and January 1998 (men). Multivariable-adjusted relative risks were calculated with Cox proportional hazards analysis. RESULTS: We ascertained 9329 events (1964 cardiovascular, 6584 cancer, and 781 other deaths) in women and 4957 events (1670 cardiovascular diseases, 2500 cancers, and 787 other deaths) in men during follow up. For men and women combined, participants in the highest quintile of total fruit and vegetable intake had a relative risk for major chronic disease of 0.95 (95% confidence interval [CI] = 0.89 to 1.01) times that of those in the lowest. Total fruit and vegetable intake was inversely associated with risk of cardiovascular disease but not with overall cancer incidence, with relative risk for an increment of five servings daily of 0.88 (95% CI = 0.81 to 0.95) for cardiovascular disease and 1.00 (95% CI = 0.95 to 1.05) for cancer. Of the food groups analyzed, green leafy vegetable intake showed the strongest inverse association with major chronic disease and cardiovascular disease. For an increment of one serving per day of green leafy vegetables, relative risks were 0.95 (95% CI = 0.92 to 0.99) for major chronic disease and 0.89 (95% CI = 0.83 to 0.96) for cardiovascular disease. CONCLUSIONS: Increased fruit and vegetable consumption was associated with a modest although not statistically significant reduction in the development of major chronic disease. The benefits appeared to be primarily for cardiovascular disease and not for cancer. PMID- 15523087 TI - Prediction of irinotecan pharmacokinetics by use of cytochrome P450 3A4 phenotyping probes. AB - BACKGROUND: Irinotecan is a topoisomerase I inhibitor that has been approved for use as a first- and second-line treatment for colorectal cancer. The response to irinotecan is variable, possibly because of interindividual variation in the expression of the enzymes that metabolize irinotecan, including cytochrome P450 3A4 (CYP3A4) and uridine diphosphate glucuronosyltransferase 1A1 (UGT1A1). We prospectively explored the relationships between CYP3A phenotype, as assessed by erythromycin metabolism and midazolam clearance, and the metabolism of irinotecan and its active metabolite SN-38. METHODS: Of the 30 white cancer patients, 27 received at least two treatments with irinotecan administered as one 90-minute infusion (dose, 600 mg) with 3 weeks between treatments, and three received only one treatment. Before the first and second treatments, patients underwent an erythromycin breath test and a midazolam clearance test as phenotyping probes for CYP3A4. Erythromycin metabolism was assessed as the area under the curve for the flux of radioactivity in exhaled CO2 within 40 minutes after administration of [N methyl-14C]erythromycin. Midazolam and irinotecan were measured by high performance liquid chromatography. Genomic DNA was isolated from blood and screened for genetic variants in CYP3A4 and UGT1A1. All statistical tests were two-sided. RESULTS: CYP3A4 activity varied sevenfold (range = 0.223%-1.53% of dose) among patients, whereas midazolam clearance varied fourfold (range = 262 1012 mL/min), although intraindividual variation was small. Erythromycin metabolism was not statistically significantly associated with irinotecan clearance (P = .090), whereas midazolam clearance was highly correlated with irinotecan clearance (r = .745, P<.001). In addition, the presence of a UGT1A1 variant with a (TA)7 repeat in the promoter (UGT1A1*28) was associated with increased exposure to SN-38 (435 ng x h/mL, 95% confidence interval [CI] = 339 to 531 ng x h/mL in patients who are homozygous for wild-type UGT1A1; 631 ng x h/mL, 95% CI = 499 to 762 ng . h/mL in heterozygous patients; and 1343 ng x h/mL, 95% CI = 0 to 4181 ng x h/mL in patients who are homozygous for UGT1A1*28) (P = .006). CONCLUSION: CYP3A4 phenotype, as assessed by midazolam clearance, is statistically significantly associated with irinotecan pharmacokinetics. Evaluation of midazolam clearance combined with UGT1A1*28 genotyping may assist with optimization of irinotecan chemotherapy. PMID- 15523088 TI - Mesenchymal stem cells: potential precursors for tumor stroma and targeted delivery vehicles for anticancer agents. AB - BACKGROUND: High concentrations of interferon beta (IFN-beta) inhibit malignant cell growth in vitro. However, the therapeutic utility of IFN-beta in vivo is limited by its excessive toxicity when administered systemically at high doses. Mesenchymal stem cells (MSC) can be used to target delivery of agents to tumor cells. We tested whether MSC can deliver IFN-beta to tumors, reducing toxicity. METHODS: Human MSC were transduced with an adenoviral expression vector carrying the human IFN-beta gene (MSC-IFN-beta cells). Flow cytometry was used to measure tumor cell proliferation among in vitro co-cultures of MSC-IFN-beta cells and human MDA 231 breast carcinoma cells or A375SM melanoma cells. We used a severe combined immunodeficiency mouse xenograft model (4-10 mice per group) to examine the effects of injected MSC-IFN-beta cells and human recombinant IFN-beta on the growth of MDA 231- and A375SM-derived pulmonary metastases in vivo and on survival. All statistical tests were two-sided. RESULTS: Co-culture of MSC-IFN beta cells with A375SM cells or MDA 231 cells inhibited tumor cell growth as compared with growth of the tumor cells cultured alone (differences in mean percentage of control cell growth: -94.0% [95% confidence interval [CI] = -81.2% to -106.8%; P<.001] and -104.8% [95% CI = -82.1% to -127.5%; P<.001], respectively). Intravenous injection of MSC-IFN-beta cells into mice with established MDA 231 or A375SM pulmonary metastases led to incorporation of MSC in the tumor architecture and, compared with untreated control mice, to prolonged mouse survival (median survival for MDA 231-injected mice: 60 and 37 days for MSC injected and control mice, respectively [difference = 23.0 days (95% CI = 14.5 to 34.0 days; P<.001]; median survival for A375SM-injected mice: 73.5 and 30.0 days for MSC-injected and control mice, respectively [difference = 43.5 days (95% CI = 37.0 to 57.5 days; P<.001]). By contrast, intravenous injection of recombinant IFN-beta did not prolong survival in the same models (median survival for MDA 231 injected mice: 41.0 and 37.0 days for IFN-beta-injected and control mice, respectively [difference = 4 days, 95% CI = -5 to 10 days; P = .308]; median survival for A375SM-injected mice: 32.0 and 30.0 days for IFN-beta-injected and control mice, respectively [difference = 2 days, 95% CI = 0 to 4.5 days; P = .059]). CONCLUSIONS: Injected MSC-IFN-beta cells suppressed the growth of pulmonary metastases, presumably through the local production of IFN-beta in the tumor microenvironment. MSC may be an effective platform for the targeted delivery of therapeutic proteins to cancer sites. PMID- 15523089 TI - Synergistic antitumor activity of histamine plus melphalan in isolated limb perfusion: preclinical studies. AB - BACKGROUND: We have previously shown how tumor response of isolated limb perfusion (ILP) with melphalan was improved when tumor necrosis factor alpha (TNF alpha) was added. Taking into account that other vasoactive drugs could also improve tumor response to ILP, we evaluated histamine (Hi) as an alternative to TNF-alpha. METHODS: We used a rat ILP model to assess the combined effects of Hi and melphalan (n = 6) on tumor regression, melphalan uptake (n = 6), and tissue histology (n = 2) compared with Hi or melphalan alone. We also evaluated the growth of BN-175 tumor cells as well as apoptosis, necrosis, cell morphology, and paracellular permeability of human umbilical vein endothelial cells (HUVECs) after Hi treatment alone and in combination with melphalan. RESULTS: The antitumor effect of the combination of Hi and melphalan in vivo was synergistic, and Hi-dependent reduction in tumor volume was blocked by H1 and H2 receptor inhibitors. Tumor regression was observed in 66% of the animals treated with Hi and melphalan, compared with 17% after treatment with Hi or melphalan alone. Tumor melphalan uptake increased and vascular integrity in the surrounding tissue was reduced after ILP treatment with Hi and melphalan compared with melphalan alone. In vitro results paralleled in vivo results. BN-175 tumor cells were more sensitive to the cytotoxicity of combined treatment than HUVECs, and Hi treatment increased the permeability of HUVECs. CONCLUSIONS: Hi in combination with melphalan in ILP improved response to that of melphalan alone through direct and indirect mechanisms. These results warrant further evaluation in the clinical ILP setting and, importantly, in organ perfusion. PMID- 15523090 TI - Impaired antigen presentation and effectiveness of combined active/passive immunotherapy for epithelial tumors. AB - BACKGROUND: Although immunization with tumor antigens can eliminate many transplantable tumors in animal models, immune effector mechanisms associated with successful immunotherapy of epithelial cancers remain undefined. METHODS: Skin from transgenic mice expressing the cervical cancer-associated tumor antigen human papillomavirus type 16 (HPV16) E6 or E7 proteins from a keratin 14 promoter was grafted onto syngeneic, non-transgenic mice. Skin graft rejection was measured after active immunization with HPV16 E7 and adoptive transfer of antigen specific T cells. Cytokine secretion of lymphocytes from mice receiving skin grafts and immunotherapy was detected by enzyme-linked immunosorbent assay, and HPV16 E7-specific memory CD8+ T cells were detected by flow cytometry and ELISPOT. RESULTS: Skin grafts containing HPV16 E6-or E7-expressing keratinocytes were not rejected spontaneously or following immunization with E7 protein and adjuvant. Adoptive transfer of E7-specific T-cell receptor transgenic CD8+ T cells combined with immunization resulted in induction of antigen-specific interferon gamma-secreting CD8+ T cells and rejection of HPV16 E7-expressing grafts. Specific memory CD8+ T cells were generated by immunotherapy. However, a further HPV16 E7 graft was rejected from animals with memory T cells only after a second E7 immunization. CONCLUSIONS: Antigen-specific CD8+ T cells can destroy epithelium expressing HPV16 E7 tumor antigen, but presentation of E7 antigen from skin is insufficient to reactivate memory CD8+ T cells induced by immunotherapy. Thus, effective cancer immunotherapy in humans may need to invoke sufficient effector as well as memory T cells. PMID- 15523091 TI - Transferrin-conjugated liposome targeting of photosensitizer AlPcS4 to rat bladder carcinoma cells. AB - BACKGROUND: The efficacy and safety of photodynamic therapy for superficial bladder cancer depend on tumor-selective accumulation of the photosensitizer. Bladder transitional-cell carcinoma cells overexpress the transferrin receptor on their surface. We examined whether transferrin-mediated liposomal targeting of the photosensitizer aluminum phthalocyanine tetrasulfonate (AlPcS4) is an effective strategy to attain tumor-selective accumulation of this compound when applied intravesically. METHODS: AlPcS4 was stably encapsulated in unconjugated liposomes (Lip-AlPcS4) or transferrin-conjugated liposomes (Tf-Lip-AlPcS4). The accumulation of free AlPcS4, Lip-AlPcS4, and Tf-Lip-AlPcS4 in human AY-27 transitional-cell carcinoma cells and in an orthotopic rat bladder tumor model was visualized by fluorescence microscopy. In vitro AlPcS4 accumulation was quantified by fluorescence measurements following drug extraction, and the photodynamic efficacy of AlPcS4 was measured in a clonogenic assay. All statistical tests were two-sided. RESULTS: AY-27 cells incubated with Tf-Lip AlPcS4 had much higher intracellular AlPcS4 levels than AY-27 cells incubated with Lip-AlPcS4 (384.1 versus 3.7 microM; difference = 380.4 microM, 95% CI = 219.4 to 541.3; P = .0095). Among rats bearing AY-27 cell-derived bladder tumors, intravesical instillation with Tf-Lip-AlPcS4 resulted in mean AlPcS4 fluorescence in tumoral tissue, normal urothelium, and submucosa/muscle of 77.9 fluorescence units (fu) (95% CI = 69.1 to 86.8 fu), 4.3 fu (95% CI = 4.0 to 4.5 fu), and 1.0 (95% CI = 0.1 to 1.9 fu), respectively, whereas instillation of free AlPcS4 resulted in nonselective accumulation throughout the whole bladder wall, and Lip AlPcS4 instillation resulted in no tissue accumulation. Photodynamic therapy of AY-27 cells incubated with Lip-AlPcS4 resulted in cell viabilities greater than 90% for all concentrations and incubation times tested; photodynamic therapy of cells incubated with 1 muM Tf-Lip-AlPcS4 or AlPcS4 resulted in cell viabilities of 0.19% (95% CI = 0.02% to 0.36%) and 1.32% (95% CI = 0.46% to 2.19%), respectively. Higher concentrations of either AlPcS4 or Tf-Lip-AlPcS4 resulted in cell kills of more than 3 logs. CONCLUSIONS: Transferrin-mediated liposomal targeting of photosensitizing drugs is a promising potential tool for photodynamic therapy of superficial bladder tumors. PMID- 15523093 TI - Anthropometry, physical activity, and endometrial cancer risk: results from the Netherlands Cohort Study. AB - Although obesity is an established risk factor for endometrial cancer, evidence linking risk to height, weight change since age 20, and physical activity is limited. In this case-cohort study, 62 573 women from The Netherlands Cohort Study on Diet and Cancer were followed up from 1986 to 1995, and 226 endometrial cancer case patients were identified. In Cox proportional hazards analyses, women 175 cm or taller had an increased risk of endometrial cancer compared with those less than 160 cm (rate ratio [RR] = 2.57, 95% confidence interval [CI] = 1.32 to 4.99). Compared with women with a body mass index (BMI; kg/m2) between 20 and 22.9, women with a BMI of 30 or greater had a higher risk (RR = 4.50, 95% CI = 2.62 to 7.72; P(trend)<.001). Moreover, BMI at age 20 and BMI gain since age 20 were positively associated with endometrial cancer risk (P(trend) = .02 and <.001, respectively). Women who spent 90 minutes per day or more doing nonoccupational physical activities had a lower risk (RR = 0.54, 95% CI = 0.34 to 0.85; P(trend) = .002) compared with those who spent less than 30 minutes per day. High BMI and low physical activity were strong and independent risk factors for endometrial cancer. PMID- 15523092 TI - Association between biallelic and monoallelic germline MYH gene mutations and colorectal cancer risk. AB - The MutY human homologue (MYH) gene encodes a member of the base excision repair pathway that is involved in repairing oxidative damage to DNA. Two germline MYH gene mutations that result in Myh proteins containing amino acid substitutions Y165C and G382D (hereafter called the Y165C and G382D mutations) are associated with adenomatous poly-posis and colorectal cancer among patients from several European poly-posis registries. We used a population-based series of 1238 colorectal cancer patients and 1255 healthy control subjects from Ontario, Canada, to examine the risk of colorectal cancer among biallelic and monoallelic germline MYH Y165C and G382D mutation carriers. The entire MYH gene coding region was screened in all MYH Y165C and G382D mutation carriers. Compared with noncarriers, biallelic and monoallelic germline MYH gene mutation carriers had an increased risk of colorectal cancer and were more likely to have first-or second degree relatives with colorectal cancer (relative risk = 1.54, 95% confidence interval = 1.10 to 2.16). The increased risk of colorectal cancer in biallelic and monoallelic MYH gene mutation carriers was not consistently associated with the development of multiple adenomatous polyps. Loss of heterozygosity in at least one of four loci in MYH was detected in eight (47%) of 17 colorectal tumors from monoallelic MYH gene mutation carriers but in only two (20%) of 10 colorectal tumors from biallelic MYH gene mutation carriers. These two MYH gene mutations may account for a substantial fraction of hereditary colorectal cancer. PMID- 15523094 TI - Re: Influence of the new AJCC breast cancer staging system on sentinel lymph node positivity and false-negative rates. PMID- 15523095 TI - Re: Science peels the onion of selenium effects on prostate carcinogenesis. PMID- 15523096 TI - REV1 accumulates in DNA damage-induced nuclear foci in human cells and is implicated in mutagenesis by benzo[a]pyrenediolepoxide. AB - The REV1 gene encodes a Y-family DNA polymerase that has been postulated to have both catalytic and structural functions in translesion replication past UV photoproducts in mammalian cells. To examine if REV1 is implicated in DNA damage tolerance mechanisms after exposure of human cells to a chemical carcinogen, we generated a plasmid expressing REV1 protein fused at its C-terminus with green fluorescent protein (GFP). In transient transfection experiments, virtually all of the transfected cells had a diffuse nuclear pattern in the absence of carcinogen exposure. In contrast, in cells exposed to benzo[a]pyrenediolepoxide, the fusion protein accumulated in a focal pattern in the nucleus in 25% of the cells, and co-localized with PCNA. These data support the idea that REV1 is present at stalled replication forks. We also examined the mutagenic response at the HPRT locus of human cells that had greatly reduced levels of REV1 mRNA due to the stable expression of gene-specific ribozymes, and compared them to wild-type cells. The mutant frequency was greatly reduced in the ribozyme-expressing cells. These data indicate that REV1 is implicated in the mutagenic DNA damage tolerance response to BPDE and support the development of strategies to target this protein to prevent such mutations. PMID- 15523097 TI - U3 snoRNP and Rrp5p associate independently with Saccharomyces cerevisiae 35S pre rRNA, but Rrp5p is essential for association of Rok1p. AB - Biogenesis of eukaryotic ribosomal subunits proceeds via a series of precursor ribonucleoprotein particles that correspond to different stages in the maturation pathway. The different pre-ribosomal particles each contain a distinct complement of non-ribosomal, trans-acting factors that are crucial for correct and efficient progress of the maturation process. Although in recent years we have gained considerable insight into the composition of the pre-ribosomal particles, our knowledge how the ordered association with and their dissociation from the pre ribosome of these trans-acting factors is controlled is still quite limited. Here, we have studied the mutual dependence between three of these factors, Rrp5p, U3 snoRNP and Rok1p, all essential for the early stages of pre-rRNA processing/assembly, for association with the 35S pre-rRNA in Saccharomyces cerevisiae. Using co-immunoprecipitation assays, we show that Rrp5p and U3 snoRNP associate independently of each other and that the two factors do not detectably interact prior to incorporation into the pre-ribosome. In contrast, association of the putative RNA helicase Rok1p, which is known to genetically interact with Rrp5p, is absolutely dependent on the presence of the latter protein but does not require U3. PMID- 15523099 TI - Reassessment of a free radical theory of cancer with emphasis on ultraviolet carcinogenesis. AB - Pro-oxidants, reactive species and free radicals, are toxic substances that can cause oxidative damage to major constituents of biological systems. In contradistinction, antioxidants are defined as any substance that significantly prevents the pro-oxidant-initiated oxidation of a substrate. Consequently, it was suggested that it might be possible to reduce free radical damage and thus cancer risk through 3 dietary changes: (1) caloric reduction, that is, lowering the level of free radical reactions arising in the course of normal metabolism; (2) minimize dietary components that increase the level of free radical reactions (eg, polyunsaturated fats); and (3) supplement the diet with one or more free radical reaction inhibitors (antioxidants). Lipid peroxidation exemplifies the type of chain reaction initiated by free radicals in (2) and (3). Both the phenolic antioxidant butylated hydroxytoluene (BHT) and the carotenoid beta carotene can terminate such reactions and have been shown to influence ultraviolet (UV) carcinogenesis. However, there is a lack of correlation between physicochemical and patho-physiological responses in both instances. Whereas the influence on UV carcinogenesis of both antioxidants has been reported to diminish as the level of dietary fat decreases, pointing to the involvement of lipid peroxidative reactions, the mode of BHT's action in inhibiting UV carcinogenesis appears to be related to UV dose diminution through increased spectral absorbance of the stratum corneum. beta-carotene has no such effect and may actually exacerbate UV carcinogenesis under certain dietary conditions. This paradox points to the complex relationship between chemical mechanisms and biological mode of action of antioxidants. Recent clinical and experimental data suggest that antioxidant supplementation of the complex and intricately balanced natural antioxidant defense system as a cancer prevention strategy will demand extreme caution. PMID- 15523100 TI - Chemotherapy-associated oxidative stress: impact on chemotherapeutic effectiveness. AB - Antineoplastic agents induce oxidative stress in biological systems. During cancer chemotherapy, oxidative stress-induced lipid peroxidation generates numerous electrophilic aldehydes that can attack many cellular targets. These products of oxidative stress can slow cell cycle progression of cancer cells and cause cell cycle checkpoint arrest, effects that may interfere with the ability of anticancer drugs to kill cancer cells. The aldehydes may also inhibit drug induced apoptosis (programmed cell death) by inactivating death receptors and inhibiting caspase activity. These effects would also diminish the efficacy of the treatment. The use of anti-oxidants during chemotherapy may enhance therapy by reducing the generation of oxidative stress-induced aldehydes. PMID- 15523101 TI - 8-oxo-dG elevated in children during leukemia treatment. AB - Changes in oxidative stress in children undergoing chemotherapy for acute lymphoblastic leukemia (ALL) have not been well documented. To determine whether the measurement of the DNA oxidized base 8-oxodeoxyguanosine (8-oxo-dG) may be a useful biomarker in this population, the authors conducted an observational study on 103 children with ALL. Blood samples were collected at diagnosis, during interim maintenance (IM), and during delayed intensification (DI). Blood mononuclear cell 8-oxo-dG, measured with an immunohistochemical method, decreased from diagnosis to IM (P = .01) and increased between IM and DI (P < .01). In a pilot study, bone marrow was also collected from 16 patients at diagnosis and after 28 days of treatment, but 8-oxo-dG remained the same. The relationship between plasma and dietary intake of antioxidants and the level of 8-oxo-dG was also explored. There was a direct relationship between the intake of vitamin E at diagnosis and bone marrow 8-oxo-dG (P = .03) and an inverse relationship between beta-carotene intake and blood 8-oxo-dG at IM (P = .03) and vitamin A in-take and blood 8-oxo-dG at DI (P = .003). Plasma vitamin C (P = .02) and total carotenoids (P = .01) were inversely related to blood 8-oxo-dG at IM. In contrast, higher plasma E/total lipid levels were associated with higher 8-oxo-dG at IM and DI (P < .01). At IM, patients with higher 8-oxo-dG had an increased risk of chemotherapy dose reduction (P = .04). In conclusion, the level of 8-oxo-dG in blood mononuclear cells decreases after the start of chemotherapy and increases during aggressive chemotherapy in children with ALL. PMID- 15523102 TI - Multiple dietary antioxidants enhance the efficacy of standard and experimental cancer therapies and decrease their toxicity. AB - Cancer patients can be divided into 3 groups: those receiving standard or experimental therapy, those who have become unresponsive to these therapies, and those in remission at risk for recurrence or a second new cancer. While impressive progress in standard cancer therapy has been made, the value of this therapy in the management of solid tumors may have reached a plateau. At present, there is no strategy to reduce the risk of recurrence of the primary tumors or of a second cancer among survivors. Patients unresponsive to standard or experimental therapies have little option except for poor quality of life for the remainder of life. Therefore, additional approaches should be developed to improve the efficacy of current management of cancer. In this review, the author proposes that an active nutritional protocol that includes high doses of multiple dietary antioxidants and their derivatives (vitamin C, alpha-tocopheryl succinate, and natural beta-carotene), but not endogenously made antioxidants (glutathione- and antioxidant enzyme-elevating agents), when administered as an adjunct to radiation therapy, chemotherapy, or experimental therapy, may improve its efficacy by increasing tumor response and decreasing toxicity. This nutritional protocol can also be used when patients become unresponsive to standard therapy or experimental therapy to improve quality of life and possibly increase the survival time. The authors also propose that after completion of standard therapy and/or experimental therapy, a maintenance nutritional protocol that contains lower doses of antioxidants and their derivatives, together with modification in diet and lifestyle, may reduce the risk of recurrence of the original tumor and development of a second cancer among survivors. Experimental data and limited human studies suggest that use of these nutritional approaches may improve oncologic outcomes and decrease toxicity. This review also discusses the reasons for the current debates regarding the use of antioxidants during radiation or chemotherapy. PMID- 15523103 TI - Fruit extract of Aegle marmelos protects mice against radiation-induced lethality. AB - The radioprotective effect of a hydroalcoholic extracted material from the fruit of Aegle marmelos (AME) was studied in mice exposed to different doses of gamma radiation. The optimum dose for radioprotection was determined by administering 0, 5, 10, 20, 40, or 80 mg/kg body weight of AME intraperitoneally (ip) once daily, consecutively for 5 days before exposure to 10 Gy of gamma radiation. A total of 20 mg/kg of AME for 5 consecutive days before irradiation was found to afford maximum protection as evidenced by the highest number of survivors after 30 days postirradiation. Animals from all groups were monitored for 30 days postirradiation for development of symptoms of radiation sickness and mortality. Treatment of mice with AME before exposure to different doses of gamma radiation reduced the severity of symptoms of radiation sickness and mortality with all exposure doses. This was accompanied by an increase in number of survivors in the AME + irradiation group when compared with the concurrent sterile physiological saline (SPS) + irradiation group. AME pretreatment protected mice against the gastrointestinal as well as bone marrow deaths, as evidenced by the greater number of survivors on day 10 or 30, respectively. LD50/30 was found to be 8.2 Gy for the SPS + irradiation group, while it was 8.8 Gy for AME + irradiation. The dose-reduction factor (DRF) was found to be 1.1 for AME + irradiation group. The acute toxicity study of AME showed that it was nontoxic up to a dose of 6 g/kg body weight, the highest drug dose that could be administered. Irradiation of animals resulted in a dose-dependent elevation in lipid peroxidation in liver, kidney, stomach, and intestine of mice. Conversely, GSH concentration declined in a dose-dependent manner. Treatment of animals with AME before irradiation caused a significant decrease in the lipid peroxidation accompanied by a significant elevation in the GSH concentration in liver, kidney, stomach, and intestine of mice determined at 31 days postirradiation. PMID- 15523104 TI - Dietary antioxidants and human cancer. AB - Epidemiological studies show that a high intake of anti-oxidant-rich foods is inversely related to cancer risk. While animal and cell cultures confirm the anticancer effects of antioxidants, intervention trials to determine their ability to reduce cancer risk have been inconclusive, although selenium and vitamin E reduced the risk of some forms of cancer, including prostate and colon cancer, and carotenoids have been shown to help reduce breast cancer risk. Cancer treatment by radiation and anticancer drugs reduces inherent antioxidants and induces oxidative stress, which increases with disease progression. Vitamins E and C have been shown to ameliorate adverse side effects associated with free radical damage to normal cells in cancer therapy, such as mucositis and fibrosis, and to reduce the recurrence of breast cancer. While clinical studies on the effect of anti-oxidants in modulating cancer treatment are limited in number and size, experimental studies show that antioxidant vitamins and some phytochemicals selectively induce apoptosis in cancer cells but not in normal cells and prevent angiogenesis and metastatic spread, suggesting a potential role for antioxidants as adjuvants in cancer therapy. PMID- 15523105 TI - Antioxidants and cancer therapy: furthering the debate. AB - The consideration of whether to use antioxidants concomitantly with chemotherapy and radiation therapy has evolved into a heated debate. This special theme issue brings together several contributors to this debate, whose perspectives enlarge our views of the questions at hand, pointing out several very relevant ideas. First, the early hypotheses of the role of antioxidants in carcinogenesis gave a simplified and often inaccurate picture of the physiological effects of specific antioxidants. Antioxidants can have protective effects that have nothing to do with oxidation; on the other hand, they can under some circumstances develop prooxidant properties and promote carcinogenesis. During treatment, however, their role is far from clear and may be either quite positive or potentially negative. A number of clinical studies have already demonstrated beneficial effects of antioxidants in ameliorating side effects of chemotherapy. More theoretical work on the chemistry of antioxidants and chemotherapy drugs suggests that antioxidants might improve therapeutic efficacy of antineoplastics by counteracting aldehydes that impede the passage of cells through the cell cycle. However, detailed clinical study also makes it clear that we are only at the very beginning of understanding the dynamics of antioxidants and oxidant damage in the body during conventional treatment. Nevertheless, research is under way on radioprotective and chemoprotective substances, some of them rooted in traditional medicine and others in our understanding of dietary antioxidants, that may eventually lead to antioxidant-based supplements that support tolerability and efficacy of treatment, without protecting tumors through interference from antineoplastic treatment. PMID- 15523108 TI - If at third you don't succeed. PMID- 15523106 TI - Targeting multiple signaling pathways as a strategy for managing prostate cancer: multifocal signal modulation therapy. AB - The aberrant behavior of cancer reflects upregulation of certain oncogenic signaling pathways that promote proliferation, inhibit apoptosis, and enable the cancer to spread and evoke angiogenesis. Theoretically, it should be feasible to decrease the activity of these pathways-or increase the activity of pathways that oppose them-with noncytotoxic agents. Since multiple pathways are dysfunctional in most cancers, and cancers accumulate new oncogenic mutations as they progress, the greatest and most durable therapeutic benefit will likely be achieved with combination regimens that address several targets. Thus, a multifocal signal modulation therapy (MSMT) of cancer is proposed. This concept has already been documented by researchers who have shown that certain combinations of signal modulators-of limited utility when administered individually-can achieve dramatic suppression of tumor growth in rodent xenograft models. The present essay attempts to guide development of MSMTs for prostate cancer. Androgen ablation is a signal-modulating measure already in standard use in the management of delocalized prostate cancer. The additional molecular targets considered here include the type 1 insulin-like growth factor receptor, the epidermal growth factor receptor, mammalian target of rapamycin, NF-kappaB, hypoxia-inducible factor-1alpha, hsp90, cyclooxygenase-2, protein kinase A type I, vascular endothelial growth factor, 5-lipoxygenase, 12-lipoxygenase, angiotensin II receptor type 1, bradykinin receptor type 1, c-Src, interleukin-6, ras, MDM2, bcl 2/bclxL, vitamin D receptor, estrogen receptor-beta, and PPAR-. Various nutrients and phytochemicals suspected to have potential utility in prostate cancer prevention and therapy, but whose key molecular targets are still unknown, might reasonably be incorporated into MSMTs for prostate cancer; these include lycopene, selenium, green tea polyphenols, genistein, and silibinin. MSMTs can be developed systematically by testing various combinations of signal-modulating agents, in concentrations that can feasibly be achieved and maintained clinically, on human prostate cancer cell lines; combinations that appear promising can then be tested in xenograft models and, ultimately, in the clinic. Some signal modulators can increase response to cytotoxic drugs by upregulating effectors of apoptosis. When MSMTs fail to raise the spontaneous apoptosis rate sufficiently to achieve tumor stasis or regression, incorporation of appropriate cytotoxic agents into the regimen may improve the clinical outcome. PMID- 15523109 TI - Distribution of circadian clock-related proteins in the cephalic nervous system of the silkworm, Bombyx mori. AB - In the circadian timing systems, input pathways transmit information on the diurnal environmental changes to a core oscillator that generates signals relayed to the body periphery by output pathways. Cryptochrome (CRY) protein participates in the light perception; period (PER), Cycle (CYC), and Doubletime (DBT) proteins drive the core oscillator; and arylalkylamines are crucial for the clock output in vertebrates. Using antibodies to CRY, PER, CYC, DBT, and arylalkylamine N acetyltransferase (aaNAT), the authors examined neuronal architecture of the circadian system in the cephalic ganglia of adult silkworms. The antibodies reacted in the cytoplasm, never in the nuclei, of specific neurons. A cluster of 4 large Ia(1) neurons in each dorsolateral protocerebrum, a pair of cells in the frontal ganglion, and nerve fibers in the corpora cardiaca and corpora allata were stained with all antibodies. The intensity of PER staining in the Ia(1) cells and in 2 to 4 adjacent small cells oscillated, being maximal late in subjective day and minimal in early night. No other oscillations were detected in any cell and with any antibody. Six small cells in close vicinity to the Ia(1) neurons coexpressed CYC-like and DBT-like, and 4 to 5 of them also coexpressed aaNATlike immunoreactivity; the PER- and CRY-like antigens were each present in separate groups of 4 cells. The CYC- and aaNAT-like antigens were further colocalized in small groups of neurons in the pars intercerebralis, at the venter of the optic tract, and in the subesophageal ganglion. Remaining antibodies reacted with similarly positioned cells in the pars intercerebralis, and the DBT antibody also reacted with the cells in the subesophageal ganglion, but antigen colocalizations were not proven. The results imply that key components of the silkworm circadian system reside in the Ia(1) neurons and that additional, hierarchically arranged oscillators contribute to overt pacemaking. The retrocerebral neurohemal organs seem to serve as outlets transmitting central neural oscillations to the hemolymph. The frontal ganglion may play an autonomous function in circadian regulations. The colocalization of aaNAT- and CYC-like antigens suggests that the enzyme is functionally linked to CYC as in vertebrates and that arylalkylamines are involved in the insect output pathway. PMID- 15523110 TI - Induction and inhibition of diapause by the same photoperiod: experimental evidence for a "double circadian oscillator clock". AB - On the southern Iberian Peninsula, the seasonal life history of the large white butterfly, Pieris brassicae, comprises 2 different photoperiodically induced developmental arrests: a hibernation diapause at photophases < 11 h and an estivation diapause at photophases > 14 h. At intermediate photophases (12 h to 13 h), the butterfly responds with a nondiapause. Combined with the experimental setup to determine photosensitivity in insects, the different photoperiodic responses at long-, intermediate-, and short-night conditions were examined to gain more insight into the time measurement mechanism in P. brassicae. The study reveals evidence for a "double circadian oscillator clock" mechanism that is based on 2 submechanisms, a "short-night determining system" and a separate "long night determining system." This conclusion was drawn from the facts that an LD 9:15 long-night induces a hibernation diapause but inhibits an estivation diapause and, conversely, that an LD 16:8 short-night inhibits a hibernation diapause but induces an estivation diapause. This opposite effect of the same photoperiod supports the argument for the existence of 2 independent targets for light-dark cycles, interpreted as 2 antagonistic time measurement systems. The existence and independence of 2 systems was further shown by differences in long night versus short-night responses regarding photosensitivity, temperature dependence, and heritable factors. The long-night measurement system is most effective in the 5th larval stage, is highly affected by temperature, and is easy to manipulate by selective inbreeding. The short-night measurement system is most effective in the 4th larval stage, is largely temperature compensated, and is not affected by experimental manipulation of the longnight measurement system. PMID- 15523111 TI - Time-lapse analysis of the circadian rhythms of conidiation and growth rate in neurospora. AB - The filamentous fungus Neurospora crassa has frequently served as a model organism for the study of circadian rhythms through its ability to form conidial spores on a daily basis. This phenomenon leaves a spatial pattern of conidiation bands along a solid surface of agar after several days of growth. Using time lapse video, the authors have quantified the rate of conidiation. They have found that conidia do not form at a specified lag time after the growth front is laid down, but rather the band region tends to simultaneously develop over a short time frame. This produces a sharp peak when the conidiation rate is plotted against time. In addition, the authors used time-lapse video to assay growth rate with greater accuracy than previously reported. It is usually assumed that Neurospora's rate of growth is constant, and this assumption of linear growth has been used extensively to determine period and phase of the conidiation circadian rhythm. The authors have confirmed an earlier report of nonlinear growth rate and have shown that the growth rate varies by a factor of about 2 with each circadian cycle. They have demonstrated that the errors in calculating times of conidiation peaks are maximally 1 to 2 h if linearity is assumed. The conidiation rate and growth rate rhythms are not apparent under conditions (using mutants or high or low temperatures) where the spatial banding rhythm is not observed. In light/dark entraining conditions, the conidiation rate and growth rate rhythms maintain the same phase relationship in different T-cycles. These data are consistent with the hypothesis that the growth rate rhythm is a consequence of the conidiation rate rhythm. PMID- 15523112 TI - Effect of vitamin A depletion on nonvisual phototransduction pathways in cryptochromeless mice. AB - Mice exhibit multiple nonvisual responses to light, including 1) photoentrainment of circadian rhythm; 2) "masking," which refers to the acute effect of light on behavior, either negative (activity suppressing) or positive (activity inducing); and 3) pupillary constriction. In mammals, the eye is the sole photosensory organ for these responses, and it contains only 2 known classes of pigments: opsins and cryptochromes. No individual opsin or cryptochrome gene is essential for circadian photoreception, gene photoinduction, or masking. Previously, the authors found that mice lacking retinol-binding protein, in which dietary depletion of ocular retinaldehyde can be achieved, had normal light signaling to the SCN, as determined by per gene photoinduction. In the present study, the authors analyzed phototransduction to the SCN in vitamin A-replete and vitamin A depleted rbp-/- and rbp-/-cry1-/-cry2-/- mice using molecular and behavioral end points. They found that vitamin A-depleted rbp-/- mice exhibit either normal photoentrainment or become diurnal. In contrast, while vitamin A-replete rbp-/ cry1-/-cry2-/- mice are light responsive (with reduced sensitivity), vitamin A depleted rbp-/-cry1-/-cry2-/- mice, which presumably lack functional opsins and cryptochromes, lose most behavioral and molecular responses to light. These data demonstrate that both cryptochromes and opsins regulate nonvisual photoresponses. PMID- 15523113 TI - Phase responses to light pulses in mice lacking functional per or cry genes. AB - The phase-resetting properties of the circadian system in mice with a functional deletion in mCry1, mCry2, mPer1, or mPer2 were studied in 2 experiments. In experiment 1, mCry1(-/-) and mCry2(-/-) mice as well as mPer1(Brdm1) and mPer2(Brdm1) mutant mice were exposed to 15-min light pulses during the 1st cycle following entrainment, either early (external time [ExT] 20) or late (ExT 4) in the subjective night. In experiment 2, a full PRC was measured for all these strains by exposure to light pulses of the same duration and intensity in free running conditions in constant darkness. Directly after entrainment (experiment 1), mPer1(Brdm1) animals did not show significant phase advances by a light pulse in the late subjective night (ExT 4), as in the study by Albrecht et al. In the same experiment, mPer2(Brdm1) mice became arrhythmic too frequently to reliably measure their phase responses. Mice with a targeted gene disruption in mCry1 or mCry2 showed increased phase delays compared to wild type after exposure to a light pulse in the early subjective night (ExT 20). Otherwise, phase shifts were not significantly affected. In free run (experiment 2), all genotypes did show phase advances and phase delays. The mPer2(Brdm1) mutant PRC was above the mPer1(Brdm1) mutant and wild-type PRC (i.e., less delayed and more advanced) at most circadian phases. The mPer1(Brdm1) mutant PRC was not distinguishable from the wildtype PRC. The mCry2(-/-) mice showed much smaller phase delays than did mCry1(-/-) mice in the subjective evening (delay phase). In general, mPer2(Brdm1) mutant mice were more accelerated by light compared to mPer1(Brdm1) and wildtype control mice, whereas mCry1(-/-) mice were more delayed by light than were mCry2( /-) mice. PMID- 15523114 TI - Phenotypic differences in reentrainment behavior and sensitivity to nighttime light pulses in siberian hamsters. AB - Spontaneous reentrainment to phase shifts of the photocycle is a fundamental property of all circadian systems. Siberian hamsters are, however, unique in this regard because most fail to reentrain when the LD cycle (16-h light/day) is phase delayed by 5 h. In the present study, the authors compared reentrainment responses in hamsters from 2 colonies. One colony descended from animals trapped in the wild more than 30 years ago (designated "nonentrainers"), and the other colony was outbred as recently as 13 years ago (designated "entrainers"). As reported previously, only 10% of hamsters from the nonentrainer colony reentrained to a 5-h phase delay of the LD cycle. By contrast, 75% of animals from the entrainer colony reentrained to the phase shift. Another goal of this study was to test the hypothesis that failure to reentrain was a consequence of light exposure during the middle of the night on the day of the 5-h phase delay. This hypothesis was tested by exposing animals to 2 h of light during the early, middle, or late part of the night and then subjecting them on the next day to a 3 h phase delay of the photocycle, which is a phase shift to which all hamsters normally reentrain. All animals from both colonies reentrained when light pulses occurred early in the night, but more animals from the entrainer colony, compared to the nonentrainer colony, reentrained when the light pulse occurred in the middle or late part of the night. The phenotypic variation in reentrainment responses is similar to the variation in photoperiodic responsiveness previously reported for these 2 colonies. Phenotypic variation in both traits is due to underlying differences in circadian organization and suggests a common genetic basis for reentrainment responses and photoperiodic responsiveness. PMID- 15523115 TI - An unusual form of phase walk in a system of coupled oscillators. AB - The authors describe an unusual form of phase walk (i.e., a progressive change in phase angle between coupled oscillators) using the 10-Hz rhythmic discharges of the inferior cardiac and vertebral postganglionic sympathetic nerves (CN and VN, respectively) in hypercapnic, baroreceptor-denervated, and vagotomized cats anesthetized with urethane. Unlike phase walk ascribable to weakened coupling (desynchronization of oscillators), the phase walk of VN 10- Hz activity relative to CN10-Hz activity 1) recurred on the time scale of the respiratory cycle, 2) was bidirectional with CN-VN phase angle increasing during expiration and decreasing during inspiration, and 3) occurred over a range equivalent to one half the period of the 10-Hz rhythm rather than a full cycle. Moreover, this form of phase walk occurred during strong coupling of the 10-Hz oscillators, as reflected by CN-VN coherence values approaching 1.0. The authors propose that the bidirectional phase walk reflects a state of strong coupling of the 10-Hz oscillators controlling the CN and VN, the angle of which is reset from cycle to cycle by the continuously changing level of activity in their respiratory inputs. In addition, the data demonstrate that frequency and amplitude modulation of sympathetic nerve discharge can be independently regulated by respiratory inputs. PMID- 15523117 TI - Current concepts in intrauterine growth restriction. AB - Regulation of fetal growth is multifactorial and complex. Diverse factors, including intrinsic fetal conditions as well as maternal and environmental factors, can lead to intrauterine growth restriction (IUGR). The interaction of these factors governs the partitioning of nutrients and rate of fetal cellular proliferation and maturation. Although IUGR is probably a physiologic adaptive response to various stimuli, it is associated with distinct short- and long-term morbidities. Immediate morbidities include those associated with prematurity and inadequate nutrient reserve, while childhood morbidities relate to impaired maturation and disrupted organ development. Potential long-term effects of IUGR are debated and explained by the fetal programming hypothesis. In formulating a comprehensive approach to the management and follow-up of the growth-restricted fetus and infant, physicians should take into consideration the etiology, timing, and severity of IUGR. In addition, they should be cognizant of the immediate perinatal response of the growth-restricted infant as well as the childhood and long-term associated morbidities. A multi disciplinary approach is imperative, including early recognition and obstetrical management of IUGR, assessment of the growth-restricted newborn in the delivery room, possible monitoring in the neonatal intensive care unit, and appropriate pediatric follow-up. Future research is necessary to establish effective preventive, diagnostic, and therapeutic strategies for IUGR, perhaps affecting the health of future generations. PMID- 15523118 TI - Serious infections associated with anticytokine therapies in the rheumatic diseases. AB - The ability to target and neutralize macrophage-derived inflammatory cytokines, particularly tumor necrosis factor-alpha (TNF-alpha), has emerged in recent years as one of the most important advances in the treatment of rheumatoid arthritis, Crohn's disease, and several other systemic inflammatory diseases. In rheumatoid arthritis, for example, these biological agents rapidly reduce signs and symptoms of joint inflammation and profoundly slow the progression of joint damage. However, data that have emerged following Food and Drug Administration approval of these agents have alerted clinicians to an increased likelihood of opportunistic infections in patients treated with these agents, particularly tuberculosis. The effect of TNF inhibition on the frequency of infection with more common bacterial pathogens is less clear. Animal models of tuberculosis and other opportunistic infections have demonstrated the importance of TNF-alpha in controlling and containing intracellular pathogens. The spectrum of infections reported to date in the setting of anti-TNF-alpha treatment is reviewed here. In addition, relevant animal data illustrating potential mechanistic roles for TNF alpha in host responses to infection are also reviewed. PMID- 15523120 TI - Creutzfeldt-Jakob disease presenting as refractory nonconvulsive status epilepticus. AB - A 70-year-old woman was admitted to the intensive care unit with refractory nonconvulsive status epilepticus. Extensive evaluation including neuroimaging and cerebrospinal fluid examination was unrevealing. Brain biopsy revealed spongiosis, and prion disease was confirmed by immunostaining, providing the diagnosis of Creutzfeldt-Jakob disease. PMID- 15523119 TI - Patterns of helium-oxygen (heliox) usage in the critical care environment. AB - The objective of this study was to describe the patterns of heliox use in critical care units of an academic medical center. The design was a prospective case series involving 7 critical care units of an academic medical center. All patients receiving heliox therapy over a 4-year period were studied, with prospective recording of patient demographics and the location, mode, indication for, and duration of heliox use. Use pattern comparisons based on anatomic location (upper vs lower airway) and age group (pediatric vs adult) were performed by alpha(2) analysis and unpaired Student t test. Eighty-nine patients, aged 17.4 +/- 20.9 years, received heliox for 30.5 +/- 44.6 hours on 92 occasions. Pediatric (0.05). Trough values on these days were 0.7 +/- 0.14 microg/mL 0.8 +/- 0.14 microg/mL and 1.0 +/- 0.21 microg/mL respectively (P > 0.05). There were no differences between the <1500 g and > 1500 g sub-groups and the < 7 days and >7 days sub-groups with respect to the corresponding peak and trough values on days 1, 3 and 7. The 95% C.I. of serum concentrations were above the MIC90 for most Enterobacteriaceae species, however the lower bound of the 95% C.I. of the mean trough levels was lower than MIC90 for Pseudomonas aeruginosa and Staphylococcus aureus. No adverse effects were observed. CONCLUSIONS: Intravenous ciprofloxacin in a dose of 10 mg/kg/dose 12 hourly is an effective treatment of neonatal sepsis, but higher doses may be required for treating Staphylococcus aureus and Pseudomonas. PMID- 15523126 TI - Benefits of non invasive ventilation. AB - Mechanical ventilation of the newborn infant has increased neonatal survival. However, this increased survival has come at the expense of increased morbidity, in the form of bronchopulmonary dysplasia, and at the cost of an expensive technology. Continuous positive airway pressure (CPAP) is accepted as conferring clinical benefit in supporting the recently extubated preterm infant and in the management of apnea of prematurity. Attention is now being drawn to physiologic and clinical evidence to support CPAP use, with or without early surfactant, as a primary treatment of hyaline membrane disease. The purpose of this review is to explore these proposed benefits of non invasive ventilation and place them in the context of current clinical evidence. PMID- 15523127 TI - Cryptorchidism: what's new? PMID- 15523128 TI - Primary vesicoureteral reflux: progress of disease, somatic growth and renal parameters. AB - Thirty children (45 units) in the age group 1 day-12 yrs with primary vesicoureteral reflux were studied prospectively and periodically assessed for renal function, scarring, grade of reflux and somatic growth parameters. Four children (6 units) with grade IV reflux underwent ureteric reimplantation. Complete resolution with medical management was seen in all 12 units of grade I III reflux and in 5 of the remaining 27 units of grade IV-V reflux over 6 months 6 yrs. This group showed highly significant improvement in height . The weight gain of the above 2 groups was statistically significant compared to those with persistent reflux. Focal defects were seen initially in 62 percent refluxing units. None of the patients showed deterioration in renal function or formation of new scars over the next 1-7 years. Three children on conservative management showed persistent growth retardation with associated breakthrough infection, hypertension, multiple renal scars and poor renal functional volume. PMID- 15523129 TI - Permanent dentition in Delhi boys of age 5-14 years. AB - Eruption pattern of permanent teeth in Delhi boys was studied in 1800 well nourished children age 5-14 years. Relationship of height, weight and sexual maturity with eruption of teeth was calculated. Earliest teeth to appear were lower first molar (5.64 years) followed by lower central incisors (6.02 years), The sequence of eruption in maxilla was first molar, incisors-central then lateral, first premolar, canine, second premolar and second molar. In mandible eruption pattern canine preceded first premolar. Eruption of teeth was significantly positively related to height, weight and sexual maturity. PMID- 15523130 TI - Uveitis and anti nuclear antibody positivity in children with juvenile idiopathic arthritis. AB - This study was conducted to determine the frequency of antinuclear antibodies (ANA) positivity and uveitis in our newly diagnosed juvenile idiopathic arthritis (JIA) patients classified according to International League Against Rheumatology (ILAR) classification criteria. Ninety-two girls and 106 boys, totally 198 children were enrolled in the study. of them 36 (18.2 percent) were found to be ANA positive. Chronic anterior uveitis was detected in 20 (10.1 percent) patients. ANA positivity was determined in 4 of the systemic JIA patients, in whom no uveitis had been detected. Twenty-five of 37 patients with oligoarticular JIA were ANA positive, in 10 of them uveitis was also diagnosed. ANA were positive in 3 of 34 patients with RF positive polyarticulat JIA, only one patient had positive ANA, and another one had uveitis. Nine patients were extended JIA and in none of them, ANA positivity or uveitis were present. Of 43 patients classified as enthesitis related arthritis (ERA), uveitis was diagnosed in 6 and there was no evidence of ANA positivity, but one had uveitis. We conclude that the incidence of ANA positivity and uveitis is low in Turkish children with JIA. PMID- 15523131 TI - Antibody levels against polioviruses in children following Pulse Polio Immunization Program. AB - This cross sectional study was performed in a tertiary level teaching hospital to evaluate and compare the antibody levels in children below 6 years who had received oral polio vaccination through Pulse Polio Immunization (PPI) with those children who had received both routine immunization as well as PPI. Detail history of polio immunization was taken. Serum samples were then collected for antibody determination by neutralization tests with standard polio viruses using Vero cell lines. Total 400 children were studied; 14 were found unvaccinated. Out of the remaining 386 (96.5%) vaccinated children, 292 (75%) had received both routine and pulse polio immunization, 68 (17%) had only PPI while 26 (6.7%) had received only routine immunization. The seropositivity was lowest for P3 and highest for P2. Overall seroprevalence for PI, P2 and P3 in vaccinated children was 89.1%, 93% and 80.6% respectively, and did not differ significantly between the three vaccinated subgroups. However, children who were immunized by both routine and PPI had higher geometric mean titers (315.5, 484.7 and 187.4 for PI, P2 and P3 respectively) when compared with those who had received only PPI (P<0.001 for each PI, P2 and P3), as well as those who had received only routine immunization with OPV (P<0.05 for PI, p<0.001 for P2, and P<0.01 for P3). Despite the reasonable immunization coverage in study population, there were 29 (7.25%) triple negative cases. Hence other causes of low seroconversion should also be considered to achieve polio free India. PMID- 15523132 TI - Non-antibiotic associated C. difficile diarrhea in a 7 week-old infant. AB - We describe a rare case of non-antibiotic associated severe C. difficile diarrhea in a 7-week-old boy. He had massive fluid loss and electrolyte imbalance. He required total parentral nutrition for 10 days and eventually recovered with oral metronidazole. Most of the reported cases in literature are associated with prior antibiotic exposure or in hospitalized patients. PMID- 15523133 TI - Neonatal lupus erythematosus. AB - We describe case report of a 45 days old male baby with neonatal lupus erythematosus, who presented with 3rd degree congenital heart block and depigmented skin lesions on face and upper part of body. Diagnosis of the baby was confirmed by anti nuclear levels and skin biopsy. PMID- 15523134 TI - Psychogenic non-epileptic seizures. AB - Psychogenic non-epileptic seizures (PNES) need to be differentiated from epileptic seizures as the management varies for both. Presence of tongue biting, falling and urinary incontinence favors a diagnosis of epileptic seizures. We report two children with features suggestive of epileptic seizures that were eventually diagnosed as PNES with the help of video-EEG monitoring. PMID- 15523135 TI - Leber's amaurosis with nephronophthisis and congenital hepatic fibrosis. AB - We describe a case of Leber's amaurosis in a one-year-old girl with unusual presentations. She presented with small clue like tachypnea and nystagmoid novement of eyeswhich when pursued revealed involvement like hepatic, renal and retina. PMID- 15523136 TI - Valproate and gum hypertrophy. PMID- 15523137 TI - Spondylocostal dysostoses. PMID- 15523138 TI - Tinea faciei. PMID- 15523139 TI - Rapid assessment of childhood disabilities through key informant approach. PMID- 15523140 TI - Acute respiratory infection with CNS excitation symptoms--consider theophylline over-dosage. PMID- 15523141 TI - Posterior urethral valves with Downs syndrome presenting as scrotal urinary sinuses. PMID- 15523142 TI - Bisphosphonate therapy in polyostotic fibrous dysplasia. PMID- 15523143 TI - Neonatal necrotizing fascitis. PMID- 15523144 TI - Hepatitis C virus infection in polytransfused thalassemic children in Pakistan. PMID- 15523145 TI - CSFC-reactive protein estimation for bedside diagnosis of pyogenic meningitis. PMID- 15523146 TI - Lateral sinus thrombosis with neurocysticercosis. PMID- 15523147 TI - Vitamin D level in slum children of Delhi. PMID- 15523148 TI - Tympanic and rectal temperatures in febrile children. PMID- 15523149 TI - Molecular biomedicine and the unraveling of complex phenotypes. PMID- 15523150 TI - Fingerprinting of common wheat cultivars with an Alw44I-based AFLP method. AB - A simplified AFLP method, based on methylation-sensitive Alw44I restriction endonuclease, has been developed and evaluated for fingerprinting 15 wheat cultivars. The selected germplasms represented groups of spring and winter wheats with and without the 1BL.1RS translocation. Ten selective primers yielded 57 markers, including 19 polymorphic bands. Three markers (15.8%) were specific to wheat carrying the 1BL.1RS translocation, thus conflicting with the frequency expected by random marker distribution (2.4%), and suggesting qualitative differences in DNA methylation among winter wheat cultivars with the 1BL.1RS translocation. Mean Dice's similarities ranged from 0.85 to 0.99, thus all cultivars could be identified by the banding profile. Winter wheat cultivars, with and without the 1BL.1RS chromosome, were slightly more similar to one another (0.959) than spring wheat cultivars (0.952). Five (9%) specific markers were obtained from cultivars Sicco, Cheyenne, Fenman, Disponent and Chinese Spring. PMID- 15523151 TI - Genetic control of aspartate aminotransferase isoenzymes in Aegilops and Triticum species. AB - Zymograms of the aspartate aminotransferase (AAT, EC 2.6.1.1) activity in leaf extracts from Aegilops and Triticum species revealed three AAT zones, denoted according to the decreasing electrophoretic mobility towards the anode as AAT-1, AAT-2 and AAT-3. The AAT activity zymograms of subcellular fractions isolated from T. aestivum seedlings made it possible to establish that the AAT-1 zone is located in the mitochondria, AAT-2 in the chloroplasts and AAT-3 in the cytoplasm. Most of the total AAT activity from wheat leaves arises from the chloroplasts and cytoplasm. The AAT-3 zone exhibited the lowest electrophoretic mobility, but 3 isoenzymes occurring within were the most visibly separated. The occurrence of a single band in this zone at the AAT-3a position (closest to the anode) for the aneuploid CS3ASDt AABBDD line (the absence of long arms of the 3rd pair of homologous chromosomes in the A genome) and at the AAT-3c position for Ae. umbellulata (genome UU), as well as three bands in the whole zone for T. durum (AABB) and T. aestivum (AABBDD) each, made it possible to evaluate the subunit composition of isoenzymes in the AAT-3 zone. The band at the AAT-3a position in the zymogram is formed from bb dimers, AAT-3b from ab and AAT-3c from aa. By comparing the distribution of isoenzyme bands intensities (the result of enzymatic activity) with the mathematical models, the frequencies of the occurrence of the a and b subunits within AAT-3 zone were evaluated. In AAT-3 from T. durum, a and b occurred at the ratio of 0.54:0.46, and in that from T. aestivum - 0.62:0.38, respectively. PMID- 15523152 TI - Multivariate approach to evaluating the fatty acid composition of seed oil in a doubled haploid population of winter oilseed rape (Brassica napus L.). AB - The fatty acid composition of oil of the zero erucic acid commercial Brassica napus L. is typical for this species. It is rich in oleic acid and contains moderate levels of linoleic and linolenic acid. For human nutrition, it is advantageous primarily to obtain the highest possible content of oleic acid and to maintain the 2:1 ratio of linoleic to linolenic acid, while preserving the average total content of saturated acids. Uni- and multivariate analyses of variance were used for evaluation of doubled haploid lines of winter oilseed rape in respect of five fatty acids: palmitic (C16:0), stearic (C18:0) oleic (C18:1), linoleic (C18:2) and linolenic (C18:3). Some proposals of studying doubled haploid (DH) lines with the use of canonical transformation were also given. In MANOVA, the five original variables (individual fatty acids) were replaced by three 'new' variables (combinations of these acids) and used to evaluate DH lines with respect to the requirements concerning the nutritional role of fatty acids. The first variable was the total content of the saturated acids (C16:0 + C18:0), the second (unchanging) was the content of the monounsaturated acid C18:1, and the third was the difference between polyunsaturated acids, i.e. between linoleic acid, and the doubled content of linolenic acid (C18:2 - 2 x C18:3). PMID- 15523153 TI - Genomic imprinting in mammals. AB - In contrast to the biallelic expression of most genes, expression of imprinted genes is monoallelic and depends on the sex of the transmitting parents. In humans it has been implicated in some developmental failures, neurodevelopmental and neurobehavioral disorders (such as Prader-Willi/Angelman, Silver-Russel or Beckwith-Wiedemann syndromes). The aim of this review is to present the phenomenon of parental imprinting as well as its molecular mechanism in various mammals. Several maternal and paternal imprinted genes and gene clusters are described. PMID- 15523154 TI - Genetic relationship between cultured populations of Pacific oyster revealed by RAPD analysis. AB - We developed random amplified polymorphic DNA (RAPD) analysis for the assessment of the genetic relationship between cultured populations of the Pacific oyster Crassostrea gigas Thunberg in Hiroshima and Goseong, the largest oyster farming areas in Japan and Korea, respectively. Of 25 arbitrary primers comprising decamer nucleotides of random sequences, polymerase chain reaction amplifications with 5 different primers gave reproducible electrophoretic patterns. A total of 49 RAPD markers were clearly identified for the Hiroshima and Goseong populations, and 46 markers were polymorphic presenting mean polymorphism rates of the respective populations at 92.29% and 93.32%. Pairwise genetic distances of each 20 individuals from these populations served to produce a UPGMA dendrogram. The dendrogram comprised two main clusters, one of which was a nested cluster including all individuals of the Hiroshima population along with 12 individuals of the Goseong population, and the other cluster included the remaining individuals of the Goseong population. Results indicate that RAPD markers are useful for the assessment of the genetic relationships between populations of the Pacific oyster and further that a significant portion of oysters imported from Korea could be genetically related to the Hiroshima population. PMID- 15523155 TI - New SSCP polymorphism within bovine STAT5A gene and its associations with milk performance traits in Black-and-White and Jersey cattle. AB - Milk protein genes expression in cows' mammary epithelial cells is regulated mostly by the action of prolactin mediated through the STAT5A transcription factor. The STAT5A gene is a potential quantitative trait locus (QTL) and genetic marker of production traits in dairy cattle. The sequence of the bovine STAT5A gene was analysed in this study to investigate if mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Ten PCR fragments representing most important functional domains of STAT5A were screened for polymorphism. Using the SSCP method a new SNP (A/G) was found, located in intron 9 at position 9501 (GenBank AJ237937). The frequencies of alleles were estimated in 186 Black-and-White cows (0.52 and 0.48 for A and G, respectively) and in 138 Jersey cows (0.58 and 0.42 for A and G, respectively). For Black-and-White cows with different STAT5A genotypes no significant associations between STAT5A genotypes and milk performance traits were found. Statistically significant differences in the first and second lactations for milk yield, fat and protein content were found in Jersey cows. Cows with the GG genotype showed the highest milk yield, while cows with genotypes AA and AG showed higher protein contents when compared to cows with the GG genotype. Interestingly, cows with genotype AG showed significantly higher protein yields in comparison to cows with the AA genotype. For fat content, cows with genotype AA showed the highest level of this trait in the 1st and 2nd lactation. Further studies are necessary to evaluate an allele substitution effect in the population of sib-families of STAT5A heterozygous bulls. PMID- 15523156 TI - New polymorphism of FASN gene in chicken. AB - Sequencing, Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) were carried out to detect polymorphism in the last intron of the FASN gene of the Campero broiler line. The analysis of the sequences presents a G to A substitution located at base 459 of the PCR product (GenBank accession number J02839, located at base Nr. 1222), resulting in a site recognized by restriction enzymes Hae III and Ava II. Also, the sequence presents a G to A substitution (located at base 603 of the PCR product and Nr. 1366 of the J02839 GenBank accession) resulting in a site recognized by restriction enzyme Pst I. Alleles and genotype frequencies were calculated for endonucleases Hae III, Ava II and Pst I for 44 broilers. PMID- 15523157 TI - Bovine mu-calpain (CAPN1) gene: new SNP within intron 14. AB - The calpain system originally comprised molecules: two Ca2+-dependent proteases, mu-calpain and m-calpain, and a third polypeptide, calpastatin, whose only known function is to inhibit the two calpains. This proteolytic system plays a key role in the tenderisation process that occurs during post-mortem storage of meat under refrigerated conditioning. Their polymorphism is examined from the point of view of their effect on corresponding production traits. The calpain genes are investigated as potential candidate genes for a quantitative trait locus (QTL) affecting meat tenderness. In this study a new single nucleotide polymorphism (SNP) was found within intron 14 of the bovine CAPN1 gene, being transition C --> T at position 4685 nt (consensus sequence - GenBank No. AF 248054), as this mutation creates a new FokI restriction site detected with PCR-RFLP analysis. This sequence fragment of the SNP position has already been deposited in the GenBank database under accession No. AY639597. The RFLP-FokI polymorphism was studied in 141 bulls of seven breeds, including the native Polish Red (PR, preserved), and Polish Black-and White (BW) breed. The frequency of alleles T and C varied between the breeds considered, the mean reaching 0.38 and 0.62, respectively. Associations between CAPN1/FokI gene polymorphism and meat production traits were studied in BW (n = 84) young bulls. In the animals of the TT genotype the lean share in valuable cuts (%) was found more favourable than in CC animals. PMID- 15523158 TI - Tamoxifen and vitamin E treatments delay symptoms in the mouse model of Niemann Pick C. AB - Niemann-Pick C disease (NPC) is an irreversible neurodegenerative disorder without current treatment. It is the result of deficient intracellular cholesterol movement. We investigated the effects of tamoxifen and vitamin E (D alpha tocopherol) treatment on patterns of weight loss and motor function in the mouse model of Niemann-Pick C disease (Npc1-/- mice). Tamoxifen has multiple metabolic effects, including reducing oxidative damage, while vitamin E primarily has this property. Npc1-/- mice were identified and treatment was initiated at an approximate age of 21 days. Tamoxifen suspended in peanut oil was administered via intraperitoneal injection (weekly, at a dose calculated to deliver 0.023 microg/g/day). Vitamin E (25 IU) was administered orally via gavage once a week. Weight loss and Rota-Rod performance were analyzed by using Kaplan-Meyer survival curves. Tamoxifen treatment by itself significantly delayed weight loss (an endpoint of neurodegeneration) in male and female mice compared to untreated controls. Motor function was evaluated by performance on a Rota-Rod. Tamoxifen maintained Rota-Rod performance for about an extra week. Vitamin E treatment significantly delayed weight loss in females only. Rota-Rod performance was maintained slightly longer in mice treated with vitamin E. Simultaneous use of both treatments did not delay weight loss longer than tamoxifen-only treatment but had a greater effect than either treatment alone on Rota-Rod performance and demonstrated a significant positive effect on the early "learning curve" portion of the Rota-Rod evaluations. We found significant but relatively small improvements in rate of disease progression by treating Npc1-/- mice with tamoxifen and/or vitamin E. Some sex differences in response and an early improvement in Rota-Rod performance suggest areas for further study. PMID- 15523159 TI - Inhibition of cyclooxygenase-2 by diallyl sulfides (DAS) in HEK 293T cells. AB - Cyclooxygenase (COX) is involved in modulating inflammatory response through the synthesis of prostaglandins. The inducible isoform of the enzyme, COX-2, is overexpressed in some malignant and premalignant lesions. Several preclinical and clinical studies have reported COX-2 inhibition as an effective strategy for chemoprevention. Nonsteroidal anitinflammatory drugs (NASIDs) such as celecoxib, are the most widely investigated COX-2 inhibitors. The oil-soluble diallyl sulfides (DAS) include monosulfides (DAMS), disulfides (DADS) and trisulfides (DATS). They were found to be effective against canine and human tumors, the mechanism of which remains unresolved. We attempted a comparative evaluation of the antiproliferative effect of DAS in HEK 293T cells. The cells were treated with increasing concentrations of DAMS, DADS and DATS. There were significant differences between the IC50 values of DAMS, DADS and DATS. RT-PCR was performed and the expression of COX-2 was compared with that of b actin. DATS inhibited COX 2 gene expression significantly stronger than DAMS and DADS. The data are suggestive of antineoplastic effect of DAS, mediated by controlling COX-2 expression. PMID- 15523160 TI - Correlation of HLA-Cw*06 allele frequency with some clinical features of psoriasis vulgaris in the population of northern Poland. AB - Psoriasis is a common skin disease with a genetic background and significant human leukocyte antigen (HLA) associations. HLA-Cw6 is the most frequently described association, particularly with psoriasis of the early onset type. Few studies of its correlation with various psoriasis clinical phenotypes and severity of the disease have been published so far; none in the Polish population. In this study 78 patients with psoriasis vulgaris were evaluated clinically and subdivided according to the age of onset and the type of psoriasis. A system of disease severity evaluation was applied to each patient. All patients and the control group (70 unrelated persons) were typed for HLA Cw*06. The results show that Cw*06 allele frequency was higher in psoriatics than in the control group. Our investigation confirms that the Cw*06 allele is positively associated with psoriasis vulgaris of the early onset type, with a positive family history and its more severe form. PMID- 15523161 TI - Subfertile couple with inv(2),inv(9) and 16qh+. AB - This case report presents two chromosomal inversions in one of partners from a subfertile couple. The woman was referred due to a spontaneous abortion in the 5th week of pregnancy. Cytogenetic examination showed that the proband's karyotype was normal: 46,XX,16qh+, as centromeric heterochromatin is thought to be clinically insignificant. However, the proband's partner occurred to be a carrier of two pericentric inversions. His karyotype was 46,XY,inv(2)(p11q13),inv(9)(p11q13). The abnormal karyotype is recognised as a possible reason of fertility problems in the investigated couple. The risk of further miscarriages is considered high, but the risk of progeny with abnormal karyotypes is rather low, as small inversions may lead to lethal recombinants. PMID- 15523162 TI - Greek stroke score, Siriraj score and Allen score in clinical diagnosis of intracerebral hemorrhage and infarct: validation and comparison study. AB - AIM: To compare Greek stroke score with available previous two stroke scores for the diagnosis of cerebral ischemia and hemorrhage in acute stroke patients, and validate the Greek stroke score. SETTING: A tertiary hospital in India. MATERIALS AND METHODS: In a prospective study acute stroke patients were evaluated with Greek stroke score, Allen score and Siriraj stroke score. Comparability (Kappa Statistics) and validity (sensitivity, specificity, negative predictive value and positive predictive value) of the Greek stroke score and previous scores were tested. RESULT: Out of the 91 patients enrolled in the study, 47 patients had cerebral infarction and 44 patients had hemorrhage by CT scan. Allen score was uncertain/equivocal in 39 patients, Siriraj Stroke score in 22 and Greek stroke score in 47 patients. Sensitivity, Specificity, positive predictive value, negative predictive value for Allen score were 0.5(95% CI:0.34,0.58), 0.94(95% CI:0.86,0.98), 0.81(95% CI:0.56,0.95), 0.78(95% CI: 0.71,0.81) for Siriraj score were 0.75(95% CI: 0.63,0.84), 0.81(95% CI: 0.71,0.89), 0.77(95% CI: 0.65,0.86), 0.78(95% CI 0.69,0.86) and for Greek Score were 0.42(95% CI: 0.23,0.53), 0.93(95% CI: 0.87,0.98), 0.71(95% CI:0.39,0.91), 0.81(95% CI:0.75,0.85) respectively. Greek stroke score was compared with previous scores using kappa statistics which revealed substantial strength of agreement between the Allen Score for certain results. CONCLUSION: The overall comparability of Greek stroke score and Allen score was better as compared to Greek stroke score and Siriraj stroke score. Greek Stroke score was more specific in diagnosing hemorrhage as compared to Siriraj score. However, all these stroke scores lack accuracy hence could not be applied safely to guide the physician in management of stroke. PMID- 15523163 TI - A prospective, randomised, double-blind study of comparative efficacy of immediate versus daily cleaning of stethoscope using 66% ethyl alcohol. AB - OBJECTIVE: Studies have demonstrated frequent contamination of stethoscope and usefulness of different disinfectants. Albeit, studies on the precise mode of cleaning and frequency of cleaning are lacking. This study was carried out to determine efficacy of 66% ethyl alcohol as disinfectant, rate of recontamination without cleaning and benefits of daily versus immediate cleaning. METHODOLOGY: Prospective, randomised, double blind study of 100 stethoscopes. Four cultures were obtained: before cleaning (Group A), immediately after cleaning with 66% ethyl alcohol (Group B), at the end of 4 days without cleaning (Group C) and at the end of 4 days after cleaning once a day (Group D). Samples were analysed using standard microbiological methods and Colony-forming unit (CFU) count and residual microorganism was computed for all the positive cultures. Medical staff was asked about the cleaning practices. Statistical analysis was carried out using 95% confidence interval and Chi-square test. RESULTS: 90% of the stethoscopes were contaminated with one or more microorganisms. Immediate cleaning and daily cleaning were associated with a significant reduction in the rate of contamination to 28% and 25% respectively. CFU count in groups B and D dropped to less than 10 in 75% and 84.7%, while the mean residual rates were 5.2% and 3.65% respectively. Groups B and D showed no statistically significant difference in terms of efficacy of disinfection. CONCLUSIONS: 66% ethyl alcohol is an effective disinfectant. The effects of immediate cleaning and cleaning once a day on residual flora on the diaphragm of stethoscope is comparable. PMID- 15523164 TI - Feasibility of "directly observed home-based twice-daily iron therapy" (DOHBIT) for management of anemia in rural patients: a pilot study. AB - BACKGROUND: Iron deficiency anemia (IDA) is a significant public health challenge in resource-poor settings, despite strong evidence for efficacy of iron supplementation. We hypothesized, modeled on the successful initiatives in disorders like tuberculosis or HIV, that directly observed therapy is an effective and practical strategy for control of IDA in such settings. OBJECTIVE: To overcome practical constraints and optimize compliance and effectiveness of iron supplementation by "Directly Observed Home-Based twice daily Iron Therapy" (DOHBIT) through village youth volunteers. DESIGN: Prospective longitudinal evaluation of DOHBIT. SETTING: A remote rural hilly hamlet with 25 families. METHODS AND MATERIAL: 100 mg elemental iron was administered twice daily under direct supervision to 33 anemic patients at home for 90 days. Outcome measures included hemoglobin rise, weight gain and side-effects. STATISTICS: Pre- and post intervention weight and hemoglobin values were compared using paired t-test. RESULTS: 29 patients completed 3-months twice-daily iron therapy without interruption (compliance 87%). There was significant increase in mean weight (43.3 +/- 6.8 kg vs. 45.1+/- 6.9 kg; P < 0.0001) as well mean hemoglobin concentration (9.5 +/- 0.9 gm% vs. 11.7 +/- 0.7 gm%; P < 0.0001) and prevalence of anemia decreased by 40% from recruitment through the 3-months therapy. In terms of side effects, occasional constipation was stated by 3 patients, transient heartburn by two and diarrhea by none. Vomiting prompted withdrawal of iron therapy in one patient. CONCLUSIONS: Providing iron supplementation as directly observed home based therapy is feasible and successful in decreasing the prevalence of anemia in resource-poor settings. PMID- 15523165 TI - An innovative technique to cut and extract loose bent Kuntscher nail. AB - Removal of a grossly deformed nail can be a serious and perplexing problem. We report an innovative technique to cut and extract a loose bent Kuntscher nail. The technique is simple and effective; can be used in peripheral hospitals where advanced gadgetries may not be present. PMID- 15523166 TI - Quality drugs in 'strip packing' constitutes an essential component of medical care rights. PMID- 15523167 TI - Organ donation in mental retardation: a clinical dilemma. PMID- 15523168 TI - Thalassemia syndromes. PMID- 15523169 TI - Serum malondialdehyde and coronary artery disease in hemodialysis patients. AB - BACKGROUND/AIMS: It has been suggested that enhanced oxidative stress participates in the acceleration of coronary artery disease (CAD) in patients with end-stage renal disease (ESRD). The aim of this study was to investigate the relationship between the level of malondialdehyde (MDA), which is a marker of lipid peroxidation, and the severity of CAD in ESRD patients. METHODS: We conducted a study of 39 hemodialysis patients (median age 58 years; 27 males and 12 females; diabetics 44%). In these patients, the predialysis serum concentrations of MDA and C-reactive protein (CRP) were measured. We performed multirow spiral computed tomography to derive coronary artery calcification (CAC) scores, as a marker of CAD severity. RESULTS: Eleven of the 39 patients had minimal CAC (28%, CAC score <10), 10 patients had mild to moderate CAC (26%, 10 400), and 18 patients had severe CAC (46%, >400). The MDA levels increased (p < 0.05) with increasing CAC category and were correlated (r = 0.35, p < 0.05) with the CAC scores. The levels of MDA also correlated with the serum concentrations of CRP and albumin (r = 0.34, p < 0.05 and r = -0.32, p < 0.05, respectively). Patients in the highest tertile of MDA compared with the other patients were over four times as likely to have severe CAC, and the highest tertile of MDA was an independent predictor of severe CAC, along with a previous cardiovascular event. CONCLUSION: An increased level of MDA, which was associated with inflammatory markers, was a predictive factor for severe CAC in ESRD patients. PMID- 15523170 TI - Maternal and fetal blood ammonia concentrations in normal term human pregnancies. AB - BACKGROUND: The current evidence on the primary source of ammonia production in the human fetoplacental unit is potentially misleading. OBJECTIVE: The aim of the present investigation was to determine the concentration of ammonia in human maternal and fetal blood at birth and to compare them with published data in late gestation sheep. METHODS: In 12 normal human pregnancies, umbilical arterial and venous and maternal venous blood was sampled, and whole blood ammonia concentrations were measured. Data from 12 pregnant sheep and fetuses from our previous studies were utilized for comparison. RESULTS: The human fetus at delivery has higher concentrations of ammonia (60-80 microM) than the late gestation fetal lamb (25-35 microM). In the human, the arterial umbilical ammonia concentration exceeds the venous umbilical concentration, indicating a net ammonia production by fetal tissues. In sheep, the venous umbilical ammonia concentration exceeds the arterial umbilical concentration, indicating the net placental ammonia production. CONCLUSIONS: In contrast to fetal lambs, human fetuses exhibit a net production of ammonia, which may reflect differences in biologic state or a species difference. PMID- 15523171 TI - Increase in serum ionized calcium during diffusive dialysis does not affect left ventricular diastolic function. AB - An increase in serum Ca2+ during hemodialysis (HD) may lead to impaired left ventricular (LV) relaxation. Since LV diastolic function assessment in dialysis patients is hampered by preload dependence of Doppler measurements, we tested the effect of HD without ultrafiltration (UF) on these measurements. Transmitral E and A velocities, and mitral annulus e and a velocities were measured in 10 patients before and after 1 h of HD without UF. Dialysate Ca2+ was 1.75 mmol/l. Serum Ca2+ after 1 h (1.31 mmol/l; 1.28-1.46 mmol/l) was higher (p = 0.002) than before HD (1.24 mmol/l; 1.09-1.32 mmol/l). E/A (0.8; 0.4-2.8) and e/a (0.7; 0.4 1.3) after 1 h were not different than E/A (0.8; 0.6-5.1) and e/a (0.7; 0.4-1.8) before HD. The increase in serum Ca2+ does not lead to a change in Doppler parameters of LV diastolic function. Changes in these parameters after combined HD-UF are related to preload, not to serum Ca2+. PMID- 15523172 TI - Natural saline-flush is sufficient to maintain patency of immobilized-urokinase double-lumen catheter used to provide temporary blood access for hemodialysis. AB - BACKGROUND: Thrombotic occlusion is a frequent complication of central venous catheters used to provide temporary blood access on hemodialysis therapy. Heparin lock is conventionally used to maintain patency of the catheter, but the necessity of heparin-lock has not been determined yet. METHODS: After the immobilized-urokinase double-lumen central venous catheter was inserted into 48 Japanese hemodialysis patients, 22 patients randomized to the heparin group received a 20-ml saline-flush, followed by 2 ml of 1,000 U/ml heparin-lock, and 26 patients randomized to the saline group received only the 20-ml saline-flush once a day for each lumen. RESULTS: Thrombotic occlusion was observed in only 1 out of 22 patients in the heparin group and 1 out of 26 patients in the saline group. No significant difference of the catheter survival was observed between the two groups (p = 0.8599). CONCLUSIONS: Natural saline-flush is sufficient for maintaining the patency of an immobilized-urokinase double-lumen central venous catheter. PMID- 15523173 TI - Hemodialysis and ADAMTS-13. PMID- 15523174 TI - Daily hemofiltration for end-stage renal disease: a feasibility and efficacy trial. AB - BACKGROUND: Although conventional hemodialysis (CHD) is currently provided primarily on an intermittent basis for patients with end-stage renal diseases (ESRD), the value of daily dialysis is well documented. Daily hemofiltration (DHF) is a convective therapy that has received little attention as an alternative to CHD. The aim of this feasibility study was to validate and assess the efficacy of DHF (6 times/week) as an alternative to CHD (3 times/week) in patients with ESRD. METHODS: In this prospective single-arm open-labeled study, DHF was administered over 4 weeks after a 1-week run-in phase period of CHD. Blood samples were drawn at baseline and on a weekly basis during the trial, and ultrafiltrate samples were collected weekly during the DHF period. The primary endpoint was to achieve a daily spKt/V of 0.40 (equivalent to a weekly stdKt/V of approximately 2.0). To achieve this goal, the DHF prescription consisted of an exchange volume of 40% of total body water. Secondary endpoints were the evaluation of blood pressure and anemia parameters, nutritional and mineral metabolism markers and health-related quality of life measures. RESULTS: Twelve patients completed the study. Mean age was 54 years, two-thirds were men, and 83% had arteriovenous fistulae. DHF delivered a mean (+/-SD) spKt/V of 0.44 +/- 0.06. This goal was reached by delivering an exchange volume of 13-14 liters, consisting of a replacement fluid volume of 11.7 +/- 1.9 l/session and net fluid loss of 1.4 +/- 0.1 l/session. Conversion from CHD to DHF resulted in marked reduction in treatment time (3.8 vs. 2 h/session, p < 0.001) and net fluid loss (2.6 vs. 1.4 l/session, p < 0.001). Calculated weekly standard Kt/V during the last week of DHF was 1.9 +/- 0.2. DHF achieved a median beta2-microglobulin (beta2-M) removal of 170 mg/session, resulting in a decrease of mid-week pre therapy beta2-M levels (48 +/- 34 vs. 36 +/- 25 mg/l, p = 0.07). Compared with CHD, DHF resulted in lower systolic (143 +/- 19 vs. 133 +/- 16 mm Hg, p = 0.002) and diastolic (79 +/- 11 vs. 72 +/- 12 mm Hg, p = 0.002) blood pressure, reduction in antihypertensive medications (2.1 +/- 0.7 vs. 1.1 +/- 0.8, p = 0.002), higher caloric intake (1,778 +/- 441 vs. 2,044 +/- 564 kcal/day, p = 0.01), higher normalized protein catabolic rate (0.88 +/- 0.17 vs. 1.00 +/- 0.35 g/kg/day, p = 0.05), higher pre-albumin levels (330 +/- 50 vs. 378 +/- 84 g/dl, p = 0.06), higher vitality (p = 0.02), improved cognitive function (p = 0.04) and less symptoms/problems (p = 0.03). Serum albumin and potassium levels remained unchanged but the serum bicarbonate level dropped during the 1-month period of DHF. CONCLUSIONS: DHF (6 times/week) over a 1-month period achieved a weekly stdKt/V of approximately 2.0, which is similar to that recommended for 3 times weekly CHD, reduced beta2-M levels, lowered blood pressure, and had some beneficial effect on nutritional parameters, and health-related quality of life measures. PMID- 15523175 TI - Health-related quality of life of hemodialysis patients in Taiwan: a multicenter study. AB - BACKGROUND/AIMS: Health-related quality of life (HRQOL) is an important determinant of treatment effectiveness in dialysis patients. To our knowledge, there are no reports evaluating HRQOL of hemodialysis (HD) in Chinese patients. The purpose of this study is to present our results about HRQOL using the 36-Item Short-Form (SF-36) questionnaire on Taiwanese hemodialysis patients. METHODS: HRQOL was measured by using the SF-36 questionnaire in 497 HD patients in five hospitals. RESULTS: The following attributes, male gender, age <50 years old, higher education level (HEL), marriage status, employment status (EPS), less comorbid medical condition (CMC), and non-diabetic patients (NDP) were all predicted on a better Physical Component Scale (PCS). Age <50 years old, body mass index >18.5, HEL, EPS and NDP were all predicted on a higher Mental Component Scale (MCS). Scales contributing to a summary measure of physical health, the PCS score was significantly lower in women (35.0 +/- 12.3) than in men (37.9 +/- 12.3). However, there was no difference in the MCS score between women and men. In multivariate analysis, age, CMC, diabetes, serum creatinine (SCr), and erythropoietin responsiveness were significant independent predictors of PCS. Diabetes, educational level, SCr, and erythropoietin responsiveness were significant independent predictors of MCS. All of the individual scales were lower in Taiwanese HD patients than in both the general Taiwanese and US population. Each of the individual scales and MCS scores were substantially lower in the Taiwan HD group than in the US HD cohort. However, the bodily pain of PCS was significantly higher in the Taiwan HD group, although the mean PCS scores for the Taiwan HD group and the US HD study participants were nearly equal at 36.3 and 36.1, respectively. CONCLUSION: The physical and mental aspects of quality of life are substantially lower for Taiwanese HD patients, except for higher bodily pain tolerance. A number of demographic and clinical characteristics have a significant impact on HRQOL in Taiwanese HD patients. PMID- 15523176 TI - Mannheim intima-media thickness consensus. AB - Intima-media thickness (IMT) is increasingly used in clinical trials as a surrogate end point for determining the success of interventions that lower risk factors for atherosclerosis. The necessity for unified criteria to distinguish early atherosclerotic plaque formation from thickening of IMT and to standardize IMT measurements is addressed in this consensus statement. Plaque is defined as a focal structure that encroaches into the arterial lumen of at least 0.5 mm or 50% of the surrounding IMT value or demonstrates a thickness of > or =1.5 mm as measured from the media-adventitia interface to the intima-lumen interface. Standard use of IMT measurements is recommended in all epidemiological and interventional trials dealing with vascular diseases to improve characterization of the population investigated. The consensus concludes that there is no need to 'treat IMT values' nor to monitor IMT values in individual patients apart from few exceptions. Although IMT has been suggested to represent an important risk marker, it does not fulfill the characteristics of an accepted risk factor. Standardized methods recommended in this consensus statement will foster homogenous data collection and analysis. This will help to improve the power of studies incorporating IMT measurements and to facilitate the merging of large databases for meta-analyses. PMID- 15523177 TI - Reduction of body temperature with paracetamol in patients with acute stroke: randomised clinical trials are needed. PMID- 15523179 TI - Action of fluoroquinolones and Linezolid on logarithmic- and stationary-phase culture of Mycobacterium tuberculosis. AB - BACKGROUND: The new challenges involved in the chemotherapy of tuberculosis make it necessary to find novel drugs, especially ones that are useful in the latent phase of the disease. METHODS: We evaluated the activity of linezolid and fluoroquinolones against logarithmic- and stationary-phase Mycobacterium tuberculosis. RESULTS: We observed that linezolid exhibits antibacterial action, although slowly, in both situations. Quinolones with an 8-methoxy group exhibit greater activity than levofloxacin in logarithmic growth phases, whereas levofloxacin exhibits greater activity in stationary-phase growth. CONCLUSION: The study of the activity of drugs against the M. tuberculosis microorganism in the latent phase is one of the most important tools available in the fight against the tuberculosis epidemic, and both linezolid and the new fluoroquinolones appear to be promising drugs. PMID- 15523180 TI - In vitro antimicrobial activity of gatifloxacin compared with other quinolones against clinical isolates from cancer patients. AB - Owing to the predominance of gram-positive pathogens in neutropenic cancer patients, newer generation quinolones with an expanded gram-positive spectrum and enhanced potency, may have a role to play for prophylaxis and/or empiric therapy in such patients. The in vitro activity of gatifloxacin was compared with that of ciprofloxacin, levofloxacin and trovafloxacin against 848 recent clinical isolates from cancer patients. Against gram-positive organisms, gatifloxacin was the most active agent tested inhibiting all Aerococcus, Listeria monocytogens, Micrococcus, Stomatococcus mucilaginous, Bacillus, and Rhodococcus equi strains at < or =2 mg/l, its designated susceptibility breakpoint. It was also very active against methicillin-susceptible staphylococci and Streptococcus spp. (including penicillin nonsusceptible Streptococcus pneumoniae and viridans streptococci). It had moderate activity against methicillin-resistant staphylococci and Enterococcus faecalis, inhibiting 68-80% of these strains at < or =2 mg/l. Gatifloxacin also had good activity against the Enterobacteriaceae (although ciprofloxacin was more potent) inhibiting >95% of isolates at < or =1 mg/l. Nonfermentative gram-negative organisms were less susceptible to all 4 agents. Gatifloxacin was very active against Acinetobacter lwoffi (MIC100 0.12 mg/l) and had moderate activity against Acinetobacter baumanii, Chryseobacterium spp., Stenotrophomonas maltophilia, Pseudomonas aeruginosa and other Pseudomonas species. Alcaligenes xylosoxidans strains were relatively resistant to all 4 agents. PMID- 15523181 TI - Good outcomes after resection of large hepatocellular carcinoma in the non cirrhotic liver. AB - BACKGROUND: The results of partial liver resection of hepatocellular carcinoma (HCC) in non-cirrhotic livers are not well known. Therefore a retrospective study was conducted. METHODS: The medical records of 180 patients with HCC were reviewed. In 40 patients (22%), HCC occurred in a non-cirrhotic liver. A detailed analysis of these patients was performed. The diagnosis HCC was based on imaging and/or percutaneous ultrasound-guided biopsy. A biopsy of the remaining liver and peroperative findings documented the absence of cirrhosis. RESULTS: Twenty-two patients underwent partial liver resection. There was no surgical mortality. The median tumour diameter in the operated patients was 10 cm. Survival rates for operated patients at 1 and 5 years were 96 and 68%, respectively. Significant factors reducing survival were portal vein thrombosis, positive lymph nodes, microscopic vascular invasion and tumour recurrence. Tumour size at the initial moment of diagnosis was not of predictive value. After surgery with curative intent disease-free interval at 1 and 5 years were 86 and 56%, respectively. CONCLUSION: In selected patients without cirrhosis, HCC can be treated successfully by surgical resection, independent of the tumour diameter, with a 5 year survival rate of 68%. PMID- 15523182 TI - Survey among surgeons on surgical treatment strategies for secondary peritonitis. AB - BACKGROUND: There is controversy about performing either a planned relaparotomy (PR) or relaparotomy on demand (ROD) in patients with secondary peritonitis. Subjective factors influencing surgeons in decision making for either surgical treatment strategy have never been studied. METHODS: All 858 surgeons of the Association of Surgeons of The Netherlands were sent a survey with 16 case vignettes simulating peritonitis patients and evaluating the preference for PR or ROD. RESULTS: Sixty-two percent of surgeons responded to the survey. Of the returned surveys, 407 were eligible for evaluation. The responding surgeons had a slight overall preference for the ROD strategy, as shown by the mean overall preference score of 5.2 (range 3.54-6.52, with a maximal score of 7). Gastrointestinal surgeons and surgeons working in regional and smaller hospitals were significantly more in favour of a ROD strategy than their counterparts. Factors significantly influencing the preference towards PR were ischaemia as aetiology and performing a primary anastomosis; as for ROD, it was small bowel as focus, local extent of contamination and the question whether abdominal closure was possible. However, there was a considerable variability in treatment decisions by surgeons. CONCLUSION: The majority of responding surgeons would make a choice for a particular treatment strategy based on peritonitis and surgical treatment characteristics. There was a slight overall preference towards the ROD strategy despite the considerable variability per case vignette. PMID- 15523183 TI - Special management of insulin lispro in continuous subcutaneous insulin infusion in young diabetic children: a randomized cross-over study. AB - OBJECTIVE: To compare the safety, efficacy and management of insulin lispro (LP) with regular human insulin (RH) in young diabetic children treated with continuous subcutaneous insulin infusion (CSII). STUDY DESIGN: 27 very young diabetic children (age 4.6 +/- 2.2 years) treated with CSII participated in an open-label, randomized cross-over multicenter study comparing 2 periods of 16 weeks of CSII with LP or RH. RESULTS: Mean daily basal rate was significantly higher during the LP period (p = 0.04). No differences were seen in changes in HbA1c levels, number of hypoglycemic events, cutaneous infections and catheter occlusions. There was no significant difference between the two treatments for preprandial and postprandial glucose values, although prandial glucose excursions tended to be lower with LP (significant at dinner, p = 0.01). Mean blood glucose levels were significantly higher at 0.00 and 3.00 a.m. during LP therapy (p < 0.05). No episode of ketoacidosis occurred during LP treatment. More parents indicated that LP made their own and the child's daily life easier (p = 0.02) and preferred LP (p = 0.01). CONCLUSIONS: LP in CSII therapy in children is safe, as effective as RH, improved postprandial excursions, met the needs of young children in their daily life well, and gained their parents' satisfaction and preference. However, a shorter duration of LP resulted in hyperglycemia during the first part of the night, which must be compensated for by increasing nocturnal basal rates during this time. PMID- 15523184 TI - Neonatal TSH levels as an index of iodine sufficiency: differences related to time of screening sampling and methodology. AB - INTRODUCTION: Current WHO guidelines consider that under adequate iodine intake <3% of newborns should have neonatal TSH levels of >5 mU/l blood when screening is performed in cord blood or at 3 days to 3 weeks of age. OBJECTIVE: To estimate whether this absolute criterion when applied to newborns older than 48 h of age and native to Buenos Aires coincides with the traditional ones (goiter and urinary iodine in school-age children (SAC)), and if the evaluation varies with either the methodology used for TSH measurements and/or the time of specimen sampling. POPULATION AND METHODS: TSH was measured by an immunofluorometric assay (IFMA) on filter paper blood spots of 186 cord blood samples, 112 babies <48 h of age and 1,500 newborns >48 h of age, and by immunoradiometric assay (IRMA) in 238 newborns. The WHO ICCIDD absolute criteria were applied to each population. Thyroid volume was assessed by direct palpation in 500 SAC, and in 100 of them urinary iodine levels were measured. RESULTS: TSH levels were >5 mU/l blood in 11.3% of the cord blood samples and in 3.6% of the samples from babies <48 h of age, suggesting mild iodine deficiency. TSH was >5 mU/l in 2.7% of newborns >48 h of age tested by IFMA (iodine sufficient) and in 30% measured by IRMA (moderate iodine insufficiency). Median urinary iodine and goiter prevalence in SAC were 143 mug/l and 4.5%, respectively, as expected in an iodine-sufficient area. conclusion: The TSH levels in Buenos Aires conform with the WHO criterion that defines iodine sufficiency. Application of this criterion, however, to cord blood samples or samples from babies <48 h old and the use of different methodologies may lead to erroneous conclusions. PMID- 15523185 TI - Prevalence of polycystic ovary syndrome in girls aged 14-18 years in Isfahan, Iran. AB - OBJECTIVE: To determine the prevalence of clinical polycystic ovary syndrome (PCOS) in 14- to 18-year-old high school girls in Isfahan, Iran. METHODS: In this cross-sectional study, 1,000 high school girls (14-18 years old) were selected by multi-stage random sampling from different high schools in Isfahan. Following physical examination, a single physician recorded the presence of hirsutism, severe acne, androgenic alopecia, menstrual dysfunction and obesity using a validated questionnaire. Clinical PCOS was diagnosed if menstrual dysfunction and clinical hyperandrogenism were detected. RESULTS: Clinical PCOS was present in 30 (3%), hirsutism in 60 (6%), menstrual dysfunction in 74 (7.4%) and severe acne in 47 (4.7%) of the population studied. CONCLUSION: The prevalence of clinical PCOS in our study population was similar to those of other studies; however, the prevalence could have been higher with hormonal assessment. PMID- 15523186 TI - Nonpeptide corticotropin-releasing hormone receptor type 1 antagonists and their applications in psychosomatic disorders. AB - Overproduction of corticotropin-releasing hormone (CRH) and stress system abnormalities are seen in psychiatric diseases such as depression, anxiety, eating disorders, and addiction. Investigations of CRH type 1 receptor (CRHR1) nonpeptide antagonists suggest therapeutic potential for treatment of these and other neuropsychiatric diseases. However, overproduction of CRH in the brain and on its periphery and disruption of the hypothalamic-pituitary-adrenal axis are also found in 'somatic' disorders. Some rare forms of Cushing's disease and related pituitary/adrenal disorders are obvious applications for CRHR1 antagonists. In addition, however, these antagonists may also be effective in treating more common somatic diseases. Patients with obesity and metabolic syndrome who often have subtle, but chronic hypothalamic-pituitary-adrenal hyperactivity, which may reflect central dysregulation of CRH and consequently glucocorticoid hypersecretion, could possibly be treated by administration of CRHR1 antagonists. Hormonal, autonomic, and immune aberrations are also present in chronic inflammatory, autoimmune, and allergic diseases, with considerable evidence linking CRH with the observed abnormalities. Furthermore, autonomic dysregulation is a prominent feature of common gastrointestinal disorders, such as irritable bowel syndrome and peptic ulcer disease. Patients with irritable bowel syndrome and other gastrointestinal disorders frequently develop altered pain perception and affective symptoms. CRH acts peripherally to modulate bowel activity both directly through the autonomic system and centrally by processing viscerosensory and visceromotor neural signals. This review presents clinical and preclinical evidence for the role of CRH in the pathophysiology of these disorders and for potential diagnostic and therapeutic applications of CRHR1 antagonists. Recognition of a dysfunctional stress system in these and other diseases will alter the understanding and treatment of 'psychosomatic' disorders. PMID- 15523191 TI - Orbital radiotherapy for thyroid-related orbitopathy. AB - PURPOSE OF REVIEW: Orbital radiotherapy has been one of the primary treatment modalities for moderate to severe thyroid-related orbitopathy. In this review the authors discuss the role of orbital radiotherapy in this disease with particular emphasis on questions raised by recent studies. RECENT FINDINGS: Despite its widespread use, until recently there have been few well-designed, randomized clinical trials on the role of orbital radiotherapy for thyroid-related orbitopathy. Although most the literature points to a favorable effect of orbital radiotherapy, recent studies have provided us with conflicting results. SUMMARY: Orbital radiotherapy remains a safe and a widely used treatment option for severe and progressive thyroid-related orbitopathy. More randomized clinical trials are needed, especially to examine the role of orbital radiotherapy for severe thyroid related orbitopathy characterized by severe congestion and/or compressive optic neuropathy with and without the use of corticosteroids. There is a wide consensus among clinicians that this subgroup of patients is the most frequent one to which orbital radiotherapy is offered. PMID- 15523192 TI - The antiphospholipid antibody syndrome: diagnostic aspects. AB - PURPOSE OF REVIEW: The antiphospholipid antibody syndrome is an important cause of arterial and/or venous thrombosis. In this review the authors discuss recent trends in the clinical and laboratory diagnosis of this syndrome that are of relevance to ophthalmologists. RECENT FINDINGS: The diagnostic criteria of antiphospholipid antibody syndrome have been reviewed and revalidated recently. Few prospective studies have been conducted to determine the prevalence of antiphospholipid antibodies in patients with retinal vascular occlusion and the spectrum of ophthalmologic findings in patients with established antiphospholipid syndrome. New antibody tests for antiphospholipid antibody syndrome are available with increased sensitivity and specificity, but lack of standardization, and interassay and interlaboratory variation makes it difficult to determine their clinical value. SUMMARY: Ophthalmologists should be familiar with the spectrum of clinical manifestation of antiphospholipid syndrome, the diagnostic criteria of this disease, and be able to carry out a basic laboratory workup for this entity. A high index of suspicion for this disorder is also important in making the diagnosis. PMID- 15523193 TI - The silent sinus syndrome: maxillary sinus atelectasis with enophthalmos and hypoglobus. AB - PURPOSE OF REVIEW: The silent sinus syndrome is a rare clinical entity of spontaneous enophthalmos and hypoglobus caused by an alteration of the normal orbital architecture and function from maxillary sinus collapse in the setting of chronic sinusitis. RECENT FINDINGS: The maxillary sinus collapse appears to result from the development of negative sinus pressure from an acquired obstruction of the maxillary sinus outflow. Patients most often present with symptoms relating to enophthalmos, although few report any symptoms of sinus disease. SUMMARY: Characteristic radiographic features of the maxillary sinus including opacification and collapse of the antral walls with inward bowing of the orbital floor are necessary for diagnosis. PMID- 15523194 TI - Orbital apex syndrome. AB - PURPOSE OF REVIEW: Visual loss from optic neuropathy and ophthalmoplegia involving multiple cranial nerves are the hallmarks of an orbital apex syndrome. Historically, the terms superior orbital fissure, orbital apex, and cavernous sinus have been used to define the anatomic locations of a disease process. However, the diagnostic evaluation and management is similar for each of these entities. The authors reviewed the literature on the diagnosis and evaluation of disorders involving the orbital apex. RECENT FINDINGS: High-resolution MRI is the preferred modality for evaluating most lesions involving the orbital apex. CT is a useful tool in the setting of trauma, to evaluate bone involvement, or when MRI is contraindicated. Although laboratory studies may be useful adjuncts in the diagnostic evaluation of lesions involving the orbital apex, surgical biopsy is often required for definitive diagnosis. SUMMARY: Orbital apex syndromes may result from a variety of inflammatory, infectious, neoplastic, iatrogenic/traumatic, and vascular conditions. A detailed history with review of systems is important in narrowing the differential diagnosis. Management is directed at the underlying cause and may be guided by surgical biopsy. Corticosteroids may be useful if an inflammatory etiology is suspected, but should be used with caution. PMID- 15523195 TI - Ocular rosacea: an update on pathogenesis and therapy. AB - PURPOSE OF REVIEW: Ocular rosacea is a common and potentially blinding eye disorder with an uncertain etiology. Therapies currently in vogue for ocular rosacea have not been rigorously studied with regards to specific indications, optimal dosing regimens, or treatment efficacy. This review will summarize the recent literature with regards to etiology and therapy of ocular rosacea, and will also examine current thinking about the parent disorder, acne rosacea. RECENT FINDINGS: Comparatively few papers on ocular rosacea were published in the past year. Recent articles on the prevalence of ocular rosacea in patients with acne rosacea suggested that between 6 and 18% of acne rosacea patients have signs or symptoms of ocular rosacea, but few cases were confirmed by an ophthalmologist. Recent articles on the pathogenesis of ocular rosacea have focused on the role of bacterial lipases, and interleukin-1alpha and matrix metalloproteinases in the blepharitis and corneal epitheliopathy, respectively. Other reports highlighted the presence of the disorder in children, and the lack of masked, placebo-controlled studies for those therapies currently in common use. SUMMARY: The epidemiology, etiology, and optimal therapy of ocular rosacea remain to be determined, and will require a more concerted effort to delineate. PMID- 15523196 TI - Ocular manifestations of graft versus host disease. AB - PURPOSE OF REVIEW: A review of the current literature regarding the ocular manifestations and management of graft versus host disease is presented. RECENT FINDINGS: A variety of systemic and topical anti-inflammatory or immunomodulatory agents, including prednisolone acetate, cyclosporine A, FK506, autologous serum, and retinoic acid show promise in controlling ocular graft versus host disease. SUMMARY: Graft versus host disease is a common manifestation of allogeneic stem cell transplantation. Ocular manifestations are found in a majority of patients and may be the presenting symptom. Ocular findings include keratoconjunctivitis sicca, pseudomembranous conjunctivitis, corneal ulceration and perforation, and microvascular retinopathy. Systemic and local therapy may be used to control ocular disease. PMID- 15523197 TI - Pharmacotherapy for diabetic retinopathy. AB - PURPOSE OF REVIEW: Diabetic retinopathy (DR) is a potentially visually devastating complication of chronic hyperglycemia and other associated systemic abnormalities. Numerous large, prospective, randomized clinical trials have delineated the current standard prevention and treatment protocols including intensive glycemic and blood pressure control and laser photocoagulation for neovascularization and clinically significant macular edema. However, despite standard intervention, vision loss from DR still occurs at an alarming rate. Thus, more recently, researchers have directed their efforts towards better understanding the microscopic changes occurring in DR to develop more effective pharmacologic prevention and treatment strategies. RECENT FINDINGS: Phase II and III clinical studies involving antivascular endothelial growth factor (VEGF) and protein kinase C (PKC) inhibitors for the management of diabetic macular edema are underway. Researchers recently found elevated pigment endothelium-derived factor (PEDF) associated with active neovascularization, a finding that counteracts prior claims of endogenous anti-angiogenic properties. Other clinical trials are underway to evaluate the efficacy of octreotide, celecoxib, and candesartan on DR. Small clinical studies have suggested beneficial treatment effects for triamcinolone acetonide, interferon alpha-2a, and supplemental oxygen; however, other studies involving losartan, vitamins C and E, and atorvastatin failed to show any benefit. SUMMARY: Over the past decade, numerous animal models have led to a more thorough understanding of the early microvascular alterations and later neovascularization and edema observed in DR. These discoveries and subsequent human clinical studies involving direct and indirect growth factor modulation, extracellular matrix alteration, vitreolysis, and alternative DR pathways including dyslipidemia, hypoxia, and sorbitol are reviewed in this manuscript. PMID- 15523198 TI - Imaging techniques for posterior uveitis. AB - PURPOSE OF REVIEW: The goal of this review is to describe the more commonly used imaging techniques and their use to identify causes of vision loss, extent of disease, and distinctive patterns associated with various causes of posterior uveitis. RECENT FINDINGS: Distinctive patterns seen with new imaging techniques and applications are being described. Ophthalmic CT, fluorescein angiography, indocyanine green angiography, and others are demonstrating inflammation and pathology in posterior uveitis. As our experience grows with these modalities, they are being used increasingly in the diagnosis and management of patients with posterior uveitis. SUMMARY: This review familiarizes the ophthalmologist with imaging in patients with inflammatory disorders of the retina and choroid. These modalities can help with the diagnosis, treatment, and monitoring of patients with uveitis. PMID- 15523199 TI - Herpes zoster virus infection. AB - PURPOSE OF REVIEW: The virology, pathophysiology, and treatment of the varicella zoster virus (VZV) have been investigated for many years now. Infection with VZV has different ramifications for people of different ages and immune status. The various aspects of VZV disease make it difficult to treat. Selected aspects of VZV disease that pertain to ocular disease are presented. RECENT FINDINGS: The risk factors for VZV disease in the different age spectrums and with concomitant immunodeficiencies have been further clarified. Studies suggest that the VZV may persist for prolonged periods on the cornea after herpes zoster ophthalmicus (HZO). Herpes Simplex Virus (HSV) or VZV may cause many cases of idiopathic uveitis with sectoral iris atrophy. The different patterns of retinal disease caused by VZV may relate to the immune status. Systemic antiviral medications for herpes zoster should be instituted within 72 hours of the rash but could be used later. Systemic antivirals combined with systemic corticosteroids improve the early quality of life in HZ patients. Postherpetic neuralgia is not prevented by early systemic antivirals or corticosteroids. Present systemic antivirals are all effective, but Famvir offers the best dosing schedule. The VZV vaccine is effective but there are some issues that suggest the need for a different vaccination regimen. SUMMARY: Further research must be performed on the clinical and therapeutic aspects of the VZV disease. Although both the vaccine and systemic antivirals have brought tremendous improvements, the disease persists. Therapy lessens but does not eliminate many of the complications. The disease may manifest in unpredictable patterns in this era of vaccination. PMID- 15523200 TI - Ocular manifestations of West Nile virus. AB - PURPOSE OF REVIEW: To review the ocular findings of patients with West Nile Virus. RECENT FINDINGS: Ocular findings have included multifocal choroiditis, vitritis, intraretinal hemorrhages, iritis, keratic precipitates, optic neuritis, branch retinal artery occlusions, and chorioretinal scarring in a case of intrauterine transmission. SUMMARY: The ocular findings associated with WNV appear to be self-limited. All the adults reported had improved vision and most had resolution of inflammation, with or without topical steroids, over the course of a few months. It may be reasonable to obtain West Nile Virus titers in any patient presenting with the above findings and systemic illness, especially from an endemic area. PMID- 15523202 TI - Bibliography. Current world literature. Neuro-ophthalmology. PMID- 15523201 TI - Drug-induced corneal complications. AB - PURPOSE OF REVIEW: To review the common corneal manifestations of systemic medications in order to describe the characteristic clinical features associated with particular systemic drugs, the indications for drug cessation, and the risks for irreversible ocular toxicity. RECENT FINDINGS: Systemic medications may reach the cornea via the tear film, aqueous humor, and limbal vasculature. The corneal changes are often the result of the underlying chemical properties of medications. Amphiphilic medications (amiodarone, chloroquine, suramin, clofazimine, etc.) may produce a drug-induced lipidosis and development of a vortex keratopathy. Antimetabolites (cytarabine) may lead to a degeneration of basal epithelial cells with formation of epithelial microcysts. Additionally, systemically administered medications and drug metabolites may lead to a stromal or endothelial deposition. Corneal changes may result in reduced visual acuity, photophobia, and ocular irritation, though these symptoms typically resolve following drug cessation. Corneal manifestations of systemic medications are often dose related, and may reflect the potential risk for lenticular or retinal changes. SUMMARY: Corneal changes secondary to systemic medications may affect all layers of the cornea. While corneal deposition is typically not an indication for drug cessation, patients receiving particular medications should be monitored for symptoms related to corneal deposition as well as for signs of irreversible ocular toxicity. PMID- 15523203 TI - Comparison of tibial rotation strength in patients' status after anterior cruciate ligament reconstruction with hamstring versus patellar tendon autografts. AB - OBJECTIVE: To determine if there are tibial rotational strength deficits in individuals treated with a semitendinosus/gracilis or bone-patella tendon-bone (BTB) autografts for ACL reconstruction. DESIGN: Retrospective. SETTING: Clinical practice and nonprofit research site. PATIENTS: 102 subjects--34 treated with BTB autograft, 34 treated with a hamstring autograft, and 34 matched controls--were evaluated at an average of 53.1 +/- 36.0 months postoperatively. MAIN OUTCOME MEASUREMENTS: Isokinetic examination at 60, 120, and 180 degrees/s to assess internal (IT) and external tibial (ET) rotation peak torque of the operated knee and contralateral knee. Internal and external tibial rotation peak torque values were evaluated for differences between gender and surgery type (2 x 3 ANOVA). RESULTS: Contrasts of IT torque across groups revealed that surgery type exhibited a significant main effect (P < 0.0001), but gender did not (P = 0.07), and there was no interaction of surgery and gender (P = 0.64). The hamstring group was weaker in IT rotation compared with the BTB and control groups (P < 0.05). No difference was noted in IT rotation torque between BTB and control groups at all test velocities. Contrasts of ET rotation torque across groups revealed that surgery type exhibited a significant main effect (P < 0.0001), but gender did not (P = 0.48). There was no interaction of surgery and gender (P = 0.98). The BTB group was significantly weaker compared with the hamstring group (P < 0.001) and the control group (P < 0.001) in ET at all test velocities. CONCLUSIONS: Residual strength deficits in IT and ET rotation following hamstring graft and BTB graft ACL reconstruction are apparent. PMID- 15523204 TI - Examination of static and dynamic postural stability in individuals with functionally stable and unstable ankles. AB - OBJECTIVE: To determine static and dynamic postural stability differences between functional ankle instability and stable ankle groups. DESIGN: Subjects were required to balance on a single leg and remain motionless for 20 seconds. After completing 3 trials, they performed a jump-landing test, which required them to jump 50% to 55% of their maximum vertical jump height. They landed on a single leg, stabilized quickly, and remained motionless for 20 seconds. SETTING: Sports Medicine Research Laboratory. PARTICIPANTS: Subjects with functional ankle instability (n = 14) who reported at least 2 sprains and "giving way" sensations at their ankle joint within the year prior to testing. Fourteen subjects with no history of ankle sprain injury were matched to subjects with functional ankle instability. MAIN OUTCOME MEASURES: Anterior/posterior and medial/lateral mean sway quantified static postural stability during single-leg stance. Dynamic postural stability was quantified with anterior/posterior and medial/lateral time to stabilization during single-leg jump landing. RESULTS: Mean sway was not significantly different between groups in the anterior/posterior (P = 0.28) and medial/lateral (P = 0.65) directions. The functional ankle instability group took significantly longer to stabilize in the anterior/posterior (3.27 +/- 0.72 seconds vs. 2.33 +/- 0.33 seconds; P < 0.001) and medial/lateral (2.48 +/- 0.50 seconds vs. 2.00 +/- 0.65 seconds; P = 0.04) directions. CONCLUSIONS: Individuals with functional ankle instability took significantly longer to stabilize than individuals with stable ankles after a single-leg jump landing. Differences between groups were not detected with mean sway measured during single-leg stance. PMID- 15523205 TI - Sport-related concussion: factors associated with prolonged return to play. AB - OBJECTIVE: To assess predictive value of concussion signs and symptoms based on return-to-play timelines. DESIGN: Physician practice study without diagnosis that includes presentation, initial and subsequent treatment, and management of concussion. SETTING: National multisite primary care sports medicine provider locations. PARTICIPANTS: Twenty-two providers at 18 sites; 101 athletes (91 men, 10 women in the following sports: 73 football, 8 basketball, 8 soccer, 3 wrestling, 2 lacrosse, 2 skiing, 5 others; 51 college, 44 high school, 4 professional, and 2 recreational). MAIN OUTCOME MEASUREMENTS: Duration of symptoms, presence of clinical signs, and time to return to play following concussion. RESULTS: One hundred one concussions were analyzed. Pearson chi2 analysis of common early and late concussion symptoms revealed statistical significance (P < 0.05) of headache >3 hours, difficulty concentrating >3 hours, any retrograde amnesia or loss of consciousness, and return to play >7 days. There appeared to be a trend in patients with posttraumatic amnesia toward poor outcome, but this was not statistically significant. CONCLUSIONS: When evaluating concussion, symptoms of headache >3 hours, difficulty concentrating >3 hours, retrograde amnesia, or loss of consciousness may indicate a more severe injury or prolonged recovery; great caution should be exercised before returning these athletes to play. PMID- 15523206 TI - Study of hematological and biochemical parameters in runners completing a standard marathon. AB - OBJECTIVE: To study hematological and biochemical parameters prospectively in runners completing a standard 42.2-km marathon run. To determine the incidence of hyponatremia in runners, and whether consumption of nonsteroidal anti inflammatory medications (NSAIDs) was associated with alterations in serum biochemical parameters. DESIGN: Observational cohort study. SETTING: City of Christchurch (New Zealand) Marathon, June 2002. PARTICIPANTS: One hundred fifty five of the 296 athletes entered in the 2002 City of Christchurch Marathon were enrolled in the study. MAIN OUTCOME MEASURES: Athletes were weighed at race registration and immediately after the race. Blood was drawn postrace for measurement of serum sodium, potassium, creatinine, and urea concentrations and for hematological analysis (hemoglobin concentration, hematocrit, leukocyte distribution). RESULTS: Complete data sets including prerace and postrace weights, and postrace hematological and biochemical analyses were collected on 134 marathon finishers. Postrace serum sodium concentrations were directly related to changes in body weight (P < 0.0001). There were no cases of biochemical or symptomatic hyponatremia. Thirteen percent of runners had taken an NSAID in the 24 hours prior to the race. Mean values for serum creatinine (P = 0.03) and serum potassium (P = 0.007) concentrations were significantly higher in runners who had taken an NSAID. No athlete who had taken an NSAID had a postrace serum creatinine concentration less than 0.09 mmol/L. Ninety-eight percent of runners had a postrace leukocytosis (mean white cell count, 18.97 b/L), of which the major component was a raised neutrophil count (mean neutrophil count, 15.69 b/L). CONCLUSIONS: This study found no cases of hyponatremia in runners completing a standard distance marathon. This finding relates to a marathon run under ideal conditions (minimal climatic stress) and in which there were fewer aid stations (every 5 km) than is common in North American marathons (every 1.6 km). Also, aggressive hydration practices were not promoted. Consumption of NSAIDs in the 24 hours prior to distance running was associated with altered renal function. PMID- 15523207 TI - Erythrocyte sickling during exercise and thermal stress. AB - OBJECTIVE: To examine effects of exercise in the heat and fluid intake on erythrocyte sickling and neutrophil activation in carriers of sickle cell trait (HbAS). DESIGN, SETTING, AND PARTICIPANTS: Six African American men (2 HbAS; 42% HbS, 4 HbAA; 20.7 +/- 0.8 years; 87.4 +/- 9.6 kg) participated in 2 randomized sessions (separate days) each consisting of 45 minutes of brisk walking (treadmill) in a hot (33 degrees C) environment. INTERVENTION: Subjects consumed no fluids or fluid for 3 hours prior to (ad libitum) and during (1.02 L) testing. MAIN OUTCOME MEASUREMENTS: Core temperature, heart rate, and perceived exertion were measured. Forearm venous blood was analyzed for percent erythrocyte sickling and plasma myeloperoxidase. RESULTS: Time-averaged heart rate (126.6 +/- 5.7 vs. 146.7 +/- 5.9 bpm; P = 0.02) and core temperature (37.6 +/- 0.1 vs. 38.1 +/- 0.1 degrees C; P < 0.05) responses were lower during fluid versus no fluid, with no statistically significant difference in perceived exertion (12.3 +/- 0.5 vs. 13.6 +/- 0.4; P = 0.06). Erythrocyte sickling progressively increased (to 3.5%-5.5%) for HbAS carriers during no fluid exercise only. No sickling was detected in HbAA subjects. Plasma myeloperoxidase responses to exercise were greater (P = 0.03) in HbAS versus HbAA. CONCLUSIONS: Fluid ingestion at a rate sufficient to offset a body weight deficit can effectively reduce erythrocyte sickling during exercise in the heat. PMID- 15523208 TI - Inability to train, recurrent infection, and selective IgM deficiency. PMID- 15523209 TI - Unusual cause of midfoot pain in a pole vaulter. PMID- 15523210 TI - Cold urticaria following an ice application: a case study. PMID- 15523211 TI - Isolated popliteus injury in a professional football player. PMID- 15523213 TI - Serum S100B protein: what does it mean during exercise? PMID- 15523218 TI - [Can the risk of recurrent alcohol abuse after liver transplantation for alcoholic cirrhosis be predicted?]. PMID- 15523219 TI - Predictive factors of alcohol relapse after orthotopic liver transplantation for alcoholic liver disease. AB - OBJECTIVES: The objective of this prospective study was to determine whether sociological and/or alcohol-related behavioral factors could be predictive of relapse after orthotopic liver transplantation for alcoholic liver disease. METHODS: Fifty-five liver-transplanted patients out of a series of 120 alcoholic cirrhotic patients were enrolled in a randomized prospective study. This study was initially designed to compare the 2 year survival in intent-to-transplant patients versus in-intent-to-use conventional treatment patients. For all patients, an identical questionnaire was completed at inclusion, and every 3 months for 5 years to collect data on alcohol-related behavior factors. RESULTS: Fifty-one patients fulfilled the criteria for the study. The mean follow-up was 35.7 months (range: 1-86). Rate of alcohol relapse was 11% at one year and 30% at 2 years. Alcohol intake above 140 g a week was declared by 11% and 22% of patients at one and 2 years, respectively. The only variable leading to a significantly lower rate of relapse was abstinence for 6 months or more before liver transplantation (23% vs 79%, P=0.0003). This variable was also significant for patients whose alcohol intake was greater than 140 g per week (P=0.003) (adjusted relative risk=5.5; 95%CI=1.3-24.5; P=0.02). Multivariate analysis (Cox model) showed that abstinence for 6 months or more before liver transplantation was the unique predictive variable. CONCLUSION: In this prospective study of 51 patients transplanted for alcoholic liver disease, abstinence before liver transplantation was the only predictive factor of alcohol relapse after liver transplantation. PMID- 15523220 TI - [Role of ursodeoxycholic acid in pediatric cholestatic disease]. PMID- 15523221 TI - [Death due to acute pancreatitis after endoscopic retrograde cholangiography]. PMID- 15523222 TI - [Celiac disease: mass screening or diagnosis in high risk populations?]. PMID- 15523223 TI - Percutaneous video-assisted necrosectomy for infected pancreatic necrosis. AB - AIMS OF THE STUDY: Percutaneous drainage of infected pancreatic necrosis is not always efficient and morbidity is high with open necrosectomy techniques. Minimally-invasive procedures have been developed to reduce this morbidity. We report our early experience with percutaneous video-assisted necrosectomy. METHODS: Among 61 patients with acute pancreatitis treated between January 2001 and February 2003, seven developed infected pancreatic necrosis. Six of these seven patients underwent percutaneous video-assisted necrosectomy after failure of radio-guided percutaneous drainage. RESULTS: One to four sessions of percutaneous video-assisted necrosectomy were required. There was no death. Sepsis control was achieved in all patients. One patient developed postoperative peritonitis due to intraoperative contamination of the peritoneal cavity. Eighteen months after the last necrosectomy, one patient developed a pseudocyst which was successfully cured by percutaneous drainage. One patient developed diabetes mellitus. CONCLUSION: Early experience in six patients has shown that percutaneous video-assisted necrosectomy is feasible, safe and efficient, in accordance with reports in the literature. Further evaluation is necessary. PMID- 15523224 TI - Pancreatic metastases: a multicentric study of 22 patients. AB - AIMS OF THE STUDY: To evaluate the diagnosis, treatment and outcome of patients with pancreatic metastases. PATIENTS AND METHODS: We retrospectively reviewed the records of patients with pancreatic metastasis managed in the Paris area between 1990 and 2000. RESULTS: The series analyzed included 22 patients, 10 men and 12 women, mean age 61 years (range: 35-76). The primary tumors were renal-cell carcinoma (N=10), colorectal cancer (N=4), lung cancer (N=4), breast cancer (N=2), cutaneous melanoma (N=1) and ileal carcinoid (N=1). The mean interval between primary treatment and presentation was 73.5 months (range: 2-151). Diagnosis was established because of clinical symptoms (N=15) or during surveillance (N=7). Computed tomography (N=19) and endoscopic ultrasound (EUS) (N=18) mainly showed solitary and hypodense/or hypoechoic masses. Histological diagnosis was obtained before surgery by EUS-guided fine needle aspiration (N=6), ultrasound-guided biopsy (N=3) or duodenoscopy (N=3). Among 10 patients with primary renal-cell carcinoma, 7 were treated by surgery. Median global survival was 33 months. Median survival was 61 months in the event of surgical treatment and 20 months in the other patients (ns). Mean survival depended on the type of primary tumor, 61 months for renal-cell carcinoma and 33 for colorectal cancer (P=0.06). CONCLUSIONS: Most pancreatic metastases develop from renal-cell carcinoma and can occur several years after nephrectomy. Histological diagnosis is often obtained before surgery. Surgical resection must be discussed as it can allow long-term survival. PMID- 15523225 TI - Incidence of gastrointestinal cancers in France. AB - AIM: Monitoring cancer incidence and time trends is essential for cancer research and health care planning. The aim of the study was to compare the incidence of gastrointestinal cancers in twelve administrative area in France to estimate the national cancer incidence during 2000 compared with the preceding 20 years. METHODS: Incidence data was provided by cancer registries and mortality data by the French national medical research institute (INSERM). The two data sets were modeled separately over the period 1988-1997 using age-cohort models. The incidence/mortality ratio obtained from these models was applied to the mortality rates of an age-cohort model of the entire population. RESULTS: The estimated number of new cases of gastrointestinal cancer was 61,465 in 2000. Colorectal cancer was the leading localization with 36,257 cases. The incidence of gastrointestinal cancers was slightly higher in northern than in southern area. Incidence of esophageal cancer was three times that of liver cancer. Variations in incidence were less marked for other localizations. The incidence of gastric and esophageal cancer in the male population decreased between 1980 and 2000, on average by slightly more than 2% per year. Incidence of other cancers increased. The number of new cases of colorectal cancer increased by 50%. The rise in the incidence of liver cancer was particularly striking, with an increase from 2000 incident cases in 1980 to nearly 6000 in 2000. CONCLUSION: For most localizations, incidence of gastrointestinal cancers displays few geographical differences in France, but there has been a striking change in incidence trends over the past 20 years. PMID- 15523226 TI - [Surgical therapy for Crohn's disease of the colon and rectum]. PMID- 15523227 TI - [Clinical management of gastrointestinal stromal tumors]. PMID- 15523228 TI - [Association of Budd-Chiari syndrome with a coeliac disease in patient native from North Africa]. AB - We report the case of a 46 year-old woman who developed a celiac disease and Budd Chiari syndrome. No etiology for the Budd-Chiari syndrome was found. The patient was from North Africa. The long term outcome is favourable with a gluten-free diet and antivitamin K treatment. PMID- 15523229 TI - [Keratoacanthoma of the anal margin]. AB - We report the case of a keratoacanthoma of the anal margin in a 34 year-old man. Initial biopsies were negative. Diagnosis was made by histological examination of the surgical specimen. Nine cases of perianal keratoacanthoma have been previously reported in the literature. Our case is remarkable because of the young age of the patient. We discuss the differential diagnosis of anal keratoacanthoma that must be distinguished from well-differentiated squamous cell carcinoma. PMID- 15523230 TI - [Common variable immunodeficiency and celiac disease]. AB - Variable immunodeficiency is frequently associated with subtotal villous atrophy, unchanged by gluten free diet. We report two cases of common variable immunodeficiency associated with chronic symptoms of malabsorption due to total villous atrophy. Symptoms of malabsorption disappeared and histological abnormalities improved after a gluten free diet. The association between celiac disease and common variable immunodeficiency should not be considered as fortuitous. Clinicians should be aware of this association and of the low sensitivity of serologic testing in this setting. PMID- 15523231 TI - [Tropical sprue: two cases in the Paris area]. AB - Tropical sprue (TS) is a postinfective tropical malabsorption that occurs in tropical countries. TS is associated with a persisting colonization of the small intestine lumen by enterotoxinogenic bacteria that cause subsequent enterocyte damage affecting all or part of the small-intestine. We report two cases of TS that occurred in inhabitants of Paris area returned from endemic areas. The first observation concerned a 76-year old woman admitted for anorexia, loss of 20 kg and anemia. The second observation concerned a 53-year old man referred for chronic diarrhea and loss of 40 kg within 4 years. In both cases, duodenal lesions consisted of subtotal and total villous atrophy with prominent infiltration of the damaged surface epithelium with lymphocytes and infiltrate of lymphocytes and plasma cells of the lamina propria. The two patients recovered under antibiotics, confirming the diagnosis of TS. PMID- 15523232 TI - [Hypermineralocorticoidism induced by very low doses of "alcohol free pastis"]. PMID- 15523233 TI - [Imatinib mesylate-induced acute cytolytic hepatitis]. PMID- 15523234 TI - [Cryptogenic cirrhosis and membranous obstruction of the retrohepatic inferior vena cava]. PMID- 15523235 TI - [Spontaneous splenic rupture secondary to cholesterol embolisms: a case study]. PMID- 15523236 TI - [Primary ileal atrophy associated with collagen colitis: clinical efficacy of corticotherapy]. PMID- 15523237 TI - [Asymptomatic stromal tumor of the colon revealed by classic Kaposi's sarcoma in an HIV negative patient]. PMID- 15523238 TI - [The Fibrotest is an ethical choice because it is a cost-effective and anti inflationistic test. Response to Jean Louis Payen and Jean Marie Combis]. PMID- 15523239 TI - [Fibrotest and development of public research]. PMID- 15523241 TI - [Performance and its effects on health]. PMID- 15523242 TI - [Assessment of out-patient treatment of varicella in children]. AB - BACKGROUND: Varicella, an almost compulsory affection in children, is complicated in 2% of cases. Some treatments such as powders and non-steroidial anti inflammatories (NSAIs) may favorise or worsen infectious complications. OBJECTIVE: To assess out-patient prescriptions in children presenting with varicella. METHODS: Three methods were used. Collection of the out-patient prescriptions in children (n=122) subsequently presenting with varicella in one of the hospitals in the North of France conducted from January to May 2003. A telephone survey was made among 50 general practitioners in the same area in March 2003. This collected not only the prescription of the last case reported, but also the intended prescriptions when confronted with a simulated case of the disease. RESULTS: Powders were prescribed in 44 to 54% of cases, NSAIs in 14 to 16%, aspirin in 2 to 4%, and oral acyclovir in 4%. The three assessment methods did not reveal any differences in the prescription of antibiotics, more frequent in the children subsequently visiting the hospital (25 vs. 6%), and the prescriptions of paracetamol. Only 38% of the prescriptions during clinical cases corresponded to the guidelines. CONCLUSION: It is crucial that practitioners be informed on the treatment of varicella and the risks of severe cutaneous surinfections enhanced by certain prescriptions. PMID- 15523243 TI - [Major digestive surgery in octogenarians. A prospective study of 54 cases]. AB - OBJECTIVE: The interest in geriatric surgery is on the increase because of the ageing of the population. Our study reviewed the results of a non- specialised unit. Method 54 octogenarians underwent digestive surgery including visceral resection. Cancer predominated the indications (80%). RESULTS: The patients exhibited cardiovascular (87%), endocrine (18.5%) or neuropsychiatric (29.6%) disorders with 75% scoring ASA III or IV. Morbidity was of 81.5% with 20% of specifically surgical complications and a 40.2% rate of cardiovascular complications. Post-surgical mortality was of 7.4% and the survival rate at 2 years was of 44.4%. The treating physicians judged that in 65% of patients the intervention had improved the initial status of the patient and had stabilised the disease in 35% of cases. The percentage of patients living at home declined from 83.3% before the intervention to 64.8% after the intervention. Only 2 out of the 9 patients having undergone stomy of the colon following colectomy continued to improve. CONCLUSION: This study underlines the interest of major surgery in octogenarians, including in units non-specialised in geriatric surgery. PMID- 15523244 TI - [Leiomyosarcoma of the broad ligament]. AB - INTRODUCTION: Leiomyosarcoma of the broad ligament is an extremely rare malignant tumor, with only 13 cases reported. OBSERVATION: A 52 year-old woman consulted for pelvic pain and metrorrhagia. Clinical examination revealed a multinodular mass on the right side of the uterus. Pelvic ultrasound revealed a heterogeneous mass. Laparotomy found a malignant tumor arising from the right broad ligament. It was independent from the proximity organs. Treatment consisted in resection and total hysterectomy with bilateral salpingo-oophorectomy. The histologic examination led to the diagnosis of leiomyosarcoma. The patient died three months later. COMMENT: This is an exceptional case. Most authors underline the poor prognosis when tumoral relapse occurs early, as was the case in our patient. PMID- 15523245 TI - [Human metapneumovirus pneumonia in an adult patient hospitalized for suspected severe acute respiratory syndrome (SARS)]. AB - INTRODUCTION: Human metapneumovirus (hMPV) is a newly identified virus responsible for occasionally severe respiratory infections, mainly observed in children. OBSERVATION: A 59-year old man was hospitalised for respiratory problems and fever 3 days after his return from China during the epidemic of severe acute respiratory syndrome (SARS). Other than fever (>38 degrees C), the patient exhibited a dry cough, myalgias, arthralgias and mild lymphopenia and the chest radiography showed an opacity on the right lower lung. Conventional virological investigations remained negative. Detection of the novel SARS associated coronavirus was negative but hMPV detection was positive. DISCUSSION: The present observation indicates that hMPV may cause respiratory symptoms mimicking initial manifestations of SARS in adults visiting epidemic countries. CONCLUSION: hMPV testing has clinical utility in adult patients with acute respiratory tract infection. PMID- 15523246 TI - [Allopurinal-induced aseptic menigitis]. PMID- 15523247 TI - [Chronic leg artery syndrome of exceptional aetiology: regarding 2 cases]. PMID- 15523248 TI - [Subacute Mediterranean visceral leishmaniosis treated with a single injection of amphotericin B]. PMID- 15523249 TI - [Isolated increase in alkaline phosphatase following ingestion of sertraline]. PMID- 15523250 TI - [Still's disease, why not?]. PMID- 15523251 TI - [Clinical and biological manifestations of adult-onset Still's disease]. AB - A TRIAD OF FEATURES: Adult onset Still's disease (ASD) is an uncommon disorder usually associating high spiking fever, evanescent skin rash constituted of small salmon pink macules, and arthritis. NUMEROUS SYSTEMIC MANIFESTATIONS: A sore throat is common and often misleading. More than 60% of the patients develop mobile and indolent lymph nodes, usually in the cervical area. Liver involvement is common and usually limited to a mild or moderate cytolysis. However, several observations of severe hepatitis have been reported justifying strict monitoring of the liver biology in these patients. Amongst the other numerous systemic manifestations that have been reported, pericarditis is common and sometimes responsible for tamponade, the pulmonary involvement may lead to an acute respiratory distress, and the rare neurological manifestations include aseptic meningitis or cranial nerve palsy. FROM A BIOLOGICAL POINT OF VIEW: The sedimentation rate is consistently elevated and there is usually a marked elevation in the polymorphonuclears. The bacteriological survey is negative as are the immunological tests. An increase in the serum level of IL-18 might be both diagnostic and prognostic. It is the increase of the serum level of ferritin and the marked decrease in its glycosylated fraction below 20% that seem to be of more potent diagnostic value. PMID- 15523252 TI - [Diagnosis, progression and prognosis, aetiology and treatment of adult-onset Still's disease]. AB - DIAGNOSTIC DOUBTS: Despite an increased awareness of the clinical features of the disease, adult onset Still's disease (AOSD) remains a diagnosis of exclusion. Many diagnostic criteria have been published, and the most popular are those proposed by Yamaguchi, even though they do not consider the presence of hyperferritinemia or the decrease of it glycosylated fraction, and they include exclusion criteria that are difficult to satisfy. UNPREDICTABLE PROGRESSION: Three evolutive forms have been described: monocyclic, intermittent with articular and/or systemic flares, chronic, usually in the form of chronic polyarthritis. Vital prognosis is sometimes compromised by severe systemic manifestations or the occurrence of amyloidosis, whilst destructive polyarthritis, particularly common in the chronic form, may compromise the function. The aetiology of AOSD remains unknown. THERAPEUTIC UNCERTAINTIES: Treatment is largely empirical in the absence of randomized clinical trials because of the rarity of the disease. Nonsteroidal anti-inflammatory drugs could be used in moderately severe forms of the disease, but they rare rarely sufficient. Around 80% of the patients require corticosteroids. Among the disease modifying drugs, methotrexate still remains the most effective. The role of the new anti-TNF molecules remains to be specified. PMID- 15523253 TI - [Imaging of dementia with magnetic resonance]. AB - The diagnostic approach of dementia has clearly improved with the progress in medical imaging, notably magnetic resonance imaging. The conventional T1 and T2 sequences or morphological imaging have demonstrated their interest in the positive and differential diagnosis of dementia, together with the more precise description of normal ageing of the brain. The ANAES (French medicines agency) proposes systematic brain imaging, notably by magnetic resonance imaging (MRI) in their practical guidelines for the diagnosis of Alzheimer's disease (http://www.anaes.fr). THE INTEREST OF CERTAIN IMAGING TECHNIQUES: The therapeutic progress in treatment of dementia implies that the different affections be recognised as early as possible. With this in mind, the functional MRI is capable of describing the damage in cases when morphological imaging is not sufficiently specific. Recent studies have reported the interest of diffusion and perfusion imaging, activation MRI and proton spectroscopy. FROM A THERAPEUTIC POINT OF VIEW: The combination of morphological and functional approaches will provide a better definition of the groups at risk in order to target current treatments and those to come. PMID- 15523254 TI - [The place of thiazolidinediones in the treatment of type 2 diabetes]. AB - SUPPLEMENTARY ORAL ANTIDIABETICS: Thiazolidinediones or glitazones are a new class of oral antidiabetics, the effects on blood sugar control of which are mediated by the sensitivity of the peripheral tissue to the effect of insulin. Hence, their effect is complementary to that of the insulin secreting agents and that of metformin, which acts by blocking the hepatic production of glucose. OTHER BENEFICIAL EFFECTS: Other than their effect on blood glucose, thiazolidinediones have a favorable influence upon the cardiovascular risk factors or markers such as blood pressure, C-reactive protein and metalloproteinase-9. They also appear to induce long term preservation of the beta-cell capital. Principle side effects are salt-water retention with a risk of decompensation of a pre-existing heart failure and weight gain with regard to the subcutaneous adipose tissue, without deleterious effects on blood glucose control. PMID- 15523255 TI - [Physical rehabilitation of patients suffering from chronic heart failure]. AB - A BENEFICIAL METHOD: Heart failure combines with peripheral vascular and muscular abnormalities that can be effectively improved by rehabilitation. The data in the literature appears to demonstrate the efficacy and excellent tolerance of such exercise. Regarding functional results and improved quality of life, rehabilitation is as equally efficient as the medical treatment that it completes. It can currently be proposed to the majority of patients exhibiting left ventricular systolic dysfunction and who are are only partially improved with medical treatment alone. MODALITIES: The rehabilitation of heart failure must, optimally, be set-up in ambulatory settings, notably within the context of a health care network. Its modalities remain to be specified in on-going studies and its impact on prognosis has to be determined. PMID- 15523256 TI - [The pre-depressive syndrome (3/4) - Medicinal treatment]. PMID- 15523257 TI - [Crossed opinions on cardiovascular pathologies]. PMID- 15523258 TI - [News on the large clinical trials in cardiology]. PMID- 15523259 TI - [Towards the optimal management in 2004 of patients with stable coronary artery disease]. PMID- 15523260 TI - [Heart failure, a disease of the elderly]. AB - INCREASING PREVALENCE OF HOSPITALISATIONS AND MORTALITY: Heart failure represents a major public health problem. Indeed, the ageing of the population and the frequency of cardiovascular risk factors explain the considerable increase in the prevalence of heart failure over the past few years. SYSTOLIC FUNCTION IS USUALLY PRESERVED: The physiopathological features of cardiovascular ageing have resulted in the high prevalence of heart failure with preserved systolic function. Hence, in patients aged over 75 presenting with heart failure, around 50% exhibit preserved ejection fraction. THE NEED FOR GERONTOLOGICAL ASSESSMENT: The prognosis of heart failure remains severe, notably in elderly, fragile patients often exhibiting several diseases. Within this context, a gerontological assessment is crucial in order to screen for concomitant diseases, the degree of the patients' dependence and the presence of "fragility". This work-up must assess the cognitive function, autonomy, somatic status, living conditions and the medico-social management of these patients. PMID- 15523261 TI - [The difficulty in diagnosing heart failure in the elderly]. AB - THREE PRINCIPLE REASONS:Confirming the existence of heart failure in an elderly patient is difficult because of the intricacy of the symptomatology with that of other diseases, the lesser willingness of the practitioner to diagnose it, and the limited access to explorations such as echocardiography. RELATIVELY UNSPECIFIC CLINICAL SIGNS:Dyspnoea, signs of low heart rate, peripheral oedema, crepitations or tachycardia are all inconstant signs or difficult to interpret. The response to a therapeutic test with diuretics is very useful. REGARDING SUPPLEMENTARY EXAMINATIONS: An echocardiography should be systematically performed in elderly patients in order to specify the type of heart failure and the extent of an eventually curable valvulopathy. Validation of the measurement of brain natriuretic peptide (BNP) in elderly patients is in progress. PMID- 15523262 TI - [Features of the treatment of heart failure in the elderly]. AB - PHARMACOLOGICAL MODIFICATIONS TO BE TAKEN INTO ACCOUNT: In elderly patients, there is a modification in the distribution columns of drugs, an alteration in glomerular filtration (doses require adaptation) and tubular function (greater sensitivity to low salt diet and diuretics), and a reduction in the hepatic elimination capacity. FOR SYSTOLIC HEART FAILURE: The choice of drugs is in practice the same as that for younger patients: diuretics, antialdosterone agents, converting enzyme inhibitors, angiotensin II receptor antagonists, beta blockers and digitalics. FOR DIASTOLIC HEART FAILURE: The therapeutic approach combines an etiologic treatment (blood pressure, myocardial ischaemia), prevention and the rapid treatment of the decompensation factors (atrial arrhythmia), a pharmacological treatment (converting enzyme inhibitors, angiotensin II receptor antagonists, and bradycardia lowering agents). Non pharmacological measures include dietary restrictions, regular physical exercise, and education for the patient and the family. PMID- 15523263 TI - [News on the obliterating artery disease of the lower limbs]. PMID- 15523264 TI - [Systolic pressure, a powerful marker of the risk of obliterating artery disease of the lower limbs]. AB - A MARKER OF PERIPHERAL ARTERIAL DISEASE: The measurement of the ankle/arm index (AAI) is a simple, rapid and inexpensive means for the assessment of distal lower limb perfusion. Its decrease reveals an alteration in the patient's arterial status. Consecutively, AAI is an excellent marker of peripheral arterial disease, well before the onset of functional consequences. A TRIPLE MARKER: Beyond the diagnosis of peripheral arterial disease, AAI provides other interests: it is not only a tool for the functional assessment of lower limb perfusion, but also a marker of local and general atherosclerosis, as well as cardiovascular events. PMID- 15523265 TI - [Non-invasive explorations in arteriopathy of the lower limbs]. AB - FOR THE DIAGNOSIS OF AN ARTERIOPATHY: The approach is basically clinical, and vascular explorations are represented by the measurement of ankle blood pressure and calculation of the pressure index, a simple test that should systematically complete the examination. THE SEARCH FOR LOCAL LESIONS OF POTENTIALLY POOR PROGNOSIS: These are principally aneurysm and stenosis of the derivation routes that should be searched for using a Doppler in order to localise them and, completed by measurement of ankle blood pressure, specify the upstream impact and the ultrasound-Doppler analysis of certain selected areas. BEFORE REVASCULARISATION: The arteriography, examination of reference until recently, has certain limits that explain its more restricted use and the widened use of ultrasound-Doppler explorations, combined with magnetic resonance imaging and scanning. PMID- 15523266 TI - [The surgical treatment of obliterating artery disease of the lower limbs today]. AB - A NEW ALTERNATIVE: Vascular surgery can benefit from the advantages of the laparoscopic method, despite major technical problems (approach and control of the aorta, clamping and suture for arterial repair). Laparoscopic surgery has been used for the treatment of occlusive atheromatous aortoiliac lesions. FROM A TECHNICAL POINT OF VIEW: The first phase of laparoscopy consists in controlling the aorta from the renal arteries up to the inferior mesenteric artery. Then the arterial repair phase starts with the introduction of the prosthesis that is sewn on the aorta assisted by laparoscopy and the branches are tunnelled up to the femoral arteries where they are anastomosed. REGARDING THE RESULTS: In 24 patients having undergone laparoscopic treatment of atheromatous aortoiliac lesions, this technique led to most encouraging results and appears to be an exciting alternative to traditional surgery in selected patients. PMID- 15523267 TI - [Treatment of carotid stenosis: angioplasty, a well thought-out choice]. AB - FOUR THERAPEUTIC CLASSES FOR SECONDARY PREVENTION: These are statins (benefit comparable to that obtained following myocardial infarction or stroke), antiplatelet drugs, basically aspirin, clopidogrel that currently has no place in first-line treatment, converting enzyme inhibitors and beta-blockers. There is currently insufficient data to recommend the routine use of vasoactive agents in patients presenting with claudication. IN THE CASE OF CRITICAL ISCHAEMIA: When faced with this serious situation, one must control the pain and the infection and, all cases, revascularisation should be attempted. Amputation should be proposed in the case of trophic disorders and gangrene. ANGIOGENESIS, A POSSIBLE ALTERNATIVE: This is defined by the budding, from existing vessels, of new vessels under the impact of growth and stabilising factors. It is possible to distinguish arteriogenesis with development of pre-existing collateral vessels, angiogenesis itself that corresponds to the development of small vessels from the large vessels and vasculogenesis, which is the formation of new vessels mediated by circulating endothelial progenitors. PMID- 15523268 TI - [Recommendations for the carotid stenosis]. PMID- 15523269 TI - [Indications for revascularisation of a carotid stenosis]. AB - TO PREVENT ISCHEMIC STROKE: Three types of measures should be taken: correction of the vascular risk factors, administration of antithrombotic agents and carotid revascularisation in patients selected on the basis of clinical and imaging criteria. CAROTID REVASCULARISATION MEASURES: The optimal indications are carotid surgery for severe symptomatic stenosis, while the benefits are marginal in severe asymptomatic stenosis and in moderate symptomatic stenosis. INDICATIONS FOR PERCUTANEOUS ANGIOPLASTY WITH PLACING OF AN ENDOPROSTHESIS: Angioplasty with endoprosthesis, a possible alternative to surgery, must be restricted to symptomatic stenosis either in randomised studies or in severe stenosis in patients in whom surgery is technically contraindicated. PMID- 15523270 TI - [Treatment for carotid stenosis: surgery, the reference]. AB - THE CONTROVERSY: The indications for carotid revascularisation are clearly codified, but the surgical treatment is debatable. The angioplasty initially proposed for non-atheromatous stenosis or for high surgical risk patients is used by some as first-line therapy, although no benefit/risk ratio in its favour has been demonstrated. ARE THERE ANY BENEFITS WITH SURGICAL TREATMENT?: The efficacy of surgery in the treatment of atheromatous carotid stenosis has been demonstrated. The interest of angioplasty The advantages expected with carotid angioplasty are the absence of cervical incision and cervical nerve lesions, together with a reduction in the duration of hospitalisation and costs. Nevertheless, there are risks inherent to the femoral puncture and endovascular propagation from the femoral to the carotid artery. Till now, the published studies have not concluded in the superiority or equivalence of angioplasty versus surgery. FOR CERTAIN SUB-GROUPS: It has not been demonstrated but it is possible that a sub-group of high-risk patients could benefit from carotid angioplasty. There are no arguments for recommending angioplasty in the treatment of carotid re-stenosis following endarterectomy or for post-radiation stenosis. ANTI-EMBOLIC TREATMENT SHOULD BE FORESEEN: Carotid angioplasty is associated with embolus, the frequency of which is 8-fold greater than that observed after endarterectomy. The systematic use of cerebral protection is therefore recommended. TODAY: The treatment of reference of carotid stenosis remains endarterectomy conducted by vascular surgeons. PMID- 15523271 TI - [Treatment of carotid stenosis: angioplasty, a reasoned choice]. AB - MASTERING THE SURGICAL ACT: Over the last decade, the technique of carotid angioplasty has progressed and it can now be considered as mastered thanks to the combination of platelet antiaggregants, miniaturisation of the material, stenting and the use of cerebral protection devices. POTENTIAL COMPLICATIONS: Notably the occurrence of cerebral embolism despite protection. Haemorrhagic complications are also possible. After a delay of 6 months, the re-stenosis rate is of around 10%. RESULTS OF RANDOMISED STUDIES: These globally demonstrate the equivalence between endoluminal treatment and surgery, notably in patients at high surgical risk. IN PRACTICE: Carotid angioplasty can be proposed in cases of severe non atheromatous stenosis localised away from the bifurcation. It is contraindicated in the case of asymptomatic atheromatous stenosis without surgical comorbidity and when the particular anatomic conditions complicate catheterism of the carotid axis. PMID- 15523272 TI - [New horizons in the management of heart failure]. PMID- 15523273 TI - [Assessment of the management of type 2 diabetes]. PMID- 15523274 TI - [Protection from stroke, updates and perspectives]. PMID- 15523275 TI - [The dawn of a new era in cardiovascular prevention]. PMID- 15523277 TI - [New concepts in cardiology]. PMID- 15523276 TI - [Hypercholesterolemia, practical guidelines]. PMID- 15523278 TI - [The artery and the myocardium, two post-infarction targets]. PMID- 15523279 TI - [Can coronary risks be further reduced?]. PMID- 15523280 TI - [Hypercholesterolemia, the future of drug associations]. PMID- 15523281 TI - [Hypertension and coronary diseases: evolution or revolution in their management?]. PMID- 15523282 TI - [A new strategy in the treatment of hypertension and its lesions on the target organs]. PMID- 15523283 TI - [Perfection is the target for the Press Medicale]. PMID- 15523284 TI - [Analysis of the articles in the Press Medicale, new procedures]. PMID- 15523285 TI - [The methods for screening pulmonary hypertension related to systemic sclerosis in France. Descriptive survey of the French Research Group on Sclerosis]. AB - CONTEXT AND OBJECTIVES: Pulmonary hypertension (PHT) represents one of the severest complications and is life-threatening for patients suffering from systemic sclerosis (SSc). In France, the modalities for screening and treating PHT related to SSc are not well codified and no consensus has been reached. We conducted a survey among physicians inscribed on the list of the French Research Group on Sclerosis (GRFS - Groupe de Recherche Francais sur la Sclerodermie) to gather information on the status of the management of PHT related to SSc. METHODS: In 2002, we sent a questionnaire to 160 physicians, members of the GRFS, to assess the epidemiology and clinical profile of SSc patients as well as the modalities of screening and management of PHT in these patients. RESULTS: Eighty eight physicians in 71 centres replied to the questionnaire. Each centre followed up a mean of 33 SSc patients, with a global distribution of 53% limited and 47% diffused SSc. These physicians saw a mean of 5 new cases of SSc per year. The patients had been referred by town practitioners (53%) or from the hospital (47%). The mean number of SSc patients with PHT was of 5.1 per physician (1.5 new SSc + PHT patients per year). Almost all the centres (65/67) who replied systematically screened for PHT in SSc patients using Doppler echocardiography a mean of every 1.3 years. For the management of the patients exhibiting PHT, the majority (41/63) of centres collaborated with a specialized unit. Around one third of the centres treated these patients with calcium channel inhibitors (82%) and/or prostacyclin (90%). All the patients were followed-up by Doppler echocardiography. The majority of the physicians (72%) were interested in a research protocol on the subject and each could have included 4 patients, i.e., a total of 160. CONCLUSION: Pulmonary hypertension, a severe complication of SSc is screened for by the physicians of the GRFS using echocardiography with a frequency similar to Who guidelines (1.3 versus once/year). PMID- 15523286 TI - Prognostic factors of hematological recovery in life-threatening nonchemotherapy drug-induced agranulocytosis. A study of 91 patients from a single center. AB - OBJECTIVES: We studied clinical factors that may influence the duration of hematological recovery to reach neutrophil counts and thus, indirectly, the prognosis in patients with life-threatening drug-induced agranulocytosis (DIA). METHODS: Using univariate and multivariate analyses with Cox's proportional hazard models, we determined the prognostic factors for hematological recovery, defined as neutrophil counts>0.5 and>1.5.10(9)/L, in 91 patients with established life-threatening DIA. RESULTS: Multivariable analysis showed that neutrophil count<0.1.10(9)/L (at diagnosis) and infection profile: severe infections or septic shock, adversely influenced the neutrophil recovery (for the two neutrophil levels). Hematopoietic growth factors were significantly associated with rapid hematological recovery (for the two neutrophil levels). Documented microbial infections and antiplatelet DIA were also associated with rapid hematological recovery (for a neutrophil count>1.5.10(9)/L). CONCLUSION: Our findings demonstrate that in life-threatening DIA, hematological recovery is mainly dependent of the neutrophil level, the type of infections and the utilization of hematopoietic growth factors. PMID- 15523287 TI - [Physical restraint in old people's homes. Evaluation of skills following specific training]. AB - AIM: The use of physical restraint in elderly patients is sometimes necessary but not without risk. We studied the prevalence and modalities of physical restraint in elderly patients in 6 old people's homes in the urban area of Toulouse, and emphasized the need for better use by the nursing staff of physical restraint in such patients with a specific training course. METHODS: Other than the prevalence of restraint, we also examined the modalities of its use in elderly patients. We studied the respect of the Anaes (Agence nationale d'accreditation et d'evaluation en sante) guidelines regarding the use of restraint in such patients, before and after an intervention that associated first a collective training course of all the staff in the old people's homes, and then specific individual training by a visiting practitioner. The information concerning the number of patients in whom restraint was used was obtained from the manager of the home or from the head nurse and was then checked by the investigator. RESULTS: The prevalence of restraint was of 22.2% (n=110 patients) at the start of the study, 15.9% following training sessions (at 6 months) and 15.5% at the end of the study (at 9 months). Skills improved regarding the benefit/risk assessment, the installation of the person restrained and the regular removal of the restraint during the day. CONCLUSION: Our strategy helped to underline this problem for the staff. The Anaes guidelines are too strict to be applied in old people's homes. One of the key points in improved management would be the presence of a coordinating physician and nurse in all the institutions caring for the elderly. PMID- 15523288 TI - [Accidental or non-accidental brain injury in infants. Prospective study of 88 cases]. AB - OBJECTIVE: To study the epidemiology of head injury (HI) in infants, the factors favouring the occurrence of a subdural haematoma (SDH), the prevalence of retinal haemorrhages (RH) and the prognostic factors, by comparing the non-accidental (NAHI) and accidental (AHI) head injuries. RH, in particular, are of fundamental value in the diagnosis of NAHI but, in the absence of systematic studies, their sensitivity and specificity for the diagnosis of the NAHI have rarely been assessed. METHOD: We prospectively collected the clinical, ophthalmologic and radiological data of HI occurring in children under 24 months old, notably by distinguishing essential macrocrania and symptomatic macrocrania of an SDH, by classifying the HI according to its severity. RESULTS: We observed 88 cases over a period of 22 months. It 28 cases it was NAHI and in the 60 others, AHI. The SDH was often correlated with the presence of retinal haemorrhages and the absence of signs of cranial impact, but not with child abuse or with essential macrocrania. The RH were of great importance in the diagnosis of NAHI; however, non-severe RH was noted in 4 cases of AHI. The neurological prognosis was essentially correlated with the initial clinical severity. CONCLUSION: Although only representing 33% of cases, child abuse was responsible for 2/3 of the deaths and for the totality of the severe morbidity in our series. The infants exhibiting perinatal problems represented an important group at risk of abuse, which justified their regular medical-social follow-up. PMID- 15523289 TI - [Asymptomatic gastric phytobezoar and anaemia due to iron deficiency revealing an autoimmune gastritis]. AB - This observation recalls that gastric phytobezoar should lead to a search for an underlying disease and that a iron deficiency can be associated and hide macrocytosis related to a vitamin B12 deficiency. CASE REPORT: A 19 year-old woman consulted for asthenia. Microcyte anaemia associated with iron deficiency was diagnosed. Upper digestive endoscopy revealed severe, totally asymptomatic phytobezoar. Biological investigations revealed a vitamin B12 deficiency, high serum gastrin level and strong positivity for gastric antiparietal anti-cell antibodies, suggestive of an autoimmune gastritis. Total immunoglobulin A deficiency was also noted. DISCUSSION: Autoimmune gastritis is responsible for megaloblastic anaemia (vitamin B12 deficiency) but can also provoke microcytic (iron-deficiency) anaemia due to insufficient absorption of the latter and related to gastric achlorhydria. Phytobezoar might also be related to achlorhydria and/or gastroparesia associated with autoimmune gastritis. Hence, autoimmune gastritis should be searched for when confronted with unexplained gastric bezoar or iron-deficiency anaemia. PMID- 15523290 TI - [Immunomodulatory effects of intravenous immunoglobulins]. AB - Intravenous immunoglobulins (IVIg) are therapeutic preparations of normal human IgG obtained from pools of more than 1000 healthy blood donors. They are currently used in the treatment of a wide range of auto-immune diseases, whether associated with auto-antibodies or auto-reactive T lymphocytes, as well as in the treatment of systemic inflammatory diseases. Several mechanisms of action have been identified during the last 20 years, including: (i) modulation of Fc receptors expression on leukocytes and endothelial cells; (ii) interaction with complement proteins; (iii) modulation of cytokines and chemokines synthesis and release; (iv) modulation of cell proliferation and apoptosis; (v) remyelinisation; (vi) neutralisation of circulating autoantibodies; (vii) selection of repertoires of B and T lymphocytes; (viii) interaction with other cell-surface molecules on lymphocytes and monocytes; (ix) corticosteroid sparing. These mechanisms of action are multiple and often intricate. However, they are still little known and further investigations are warranted. PMID- 15523291 TI - [Hereditary intermittant fever]. AB - Other than familial mediterranean fever: Four hereditary diseases presenting in the form of intermittent inflammatory flares are now recognized and have been characterised clinically and genetically. At the head of this group is Familial Mediterranean Fever (FMF), which affects thousands of patients originating from the Mediterranean area. However the familial Mediterranean Fever is no longer the only recurrent hereditary inflammatory disease. Three other entities have now been clearly defined: intermittent fever secondary to mutations in the type 1A Tumour Necrosis Factor receptor (TNF), of dominant autosomic genetic transmission, the hyperimmunoglobulinemia D syndrome and an entity regrouping the Muckle Wells syndrome, familial cold-induced urticaria, and the Chronic Infantile Neurological Cutaneous and Articular (CINCA) syndrome. IN PRACTICE: Because they require specific management and treatment, precise diagnosis of these entities is crucial. PMID- 15523292 TI - [The interest of statins in nephrology]. AB - STATINS IN THE CASE OF CHRONIC RENAL DISEASE: Data of several large clinical trials in the general population demonstrated that hydroxymethyl glutaryl coenzyme A reductase inhibitors (statins) are effective in cardiovascular disease prevention with a relatively safe profile. Patients with chronic kidney disease (CKD) are at high risk for developing premature cardiovascular disease, so the benefits of statin therapy might be expected to be substantial in this population. Adjusted dose of statins to calcinurine inhibitors and renal function seem to exhibit a favorable risk/benefit ratio in CKD patients. STUDY RESULTS: Statin use is CKD patients has been associated with a certain efficacy of cardiovascular disease prevention in several uncontrolled trials, and one randomized trial in renal transplant recipients. Several other large-scale randomized trials in CKD patients [4D (atorvastatin), AURORA (rosuvastatin) and SHARP (simvastatin/ezetimib) are currently underway. The results of these trials will permit evidence-based medicinal arguments justifying life-long clinical use of statins for cardiovascular prevention in CKD patients with progressive renal dysfunction, but data are inconclusive. OTHER POSSIBLE EFFECTS: Clinically relevant plethoric effects associated with statin therapy in CKD patients might be restricted to the decrease of inflammation and oxidative stress. PMID- 15523293 TI - [Corticosteroid-induced osteoporosis]. AB - A FREQUENT PRESCRIPTION: Corticosteroids are widely prescribed in the long-term treatment of various pathologies (chronic inflammatory rheumatic diseases, asthma and allergic diseases, auto-immune diseases...). CONSEQUENCE OF BONE LOSS: Corticosteroids can be responsible for side effects such as corticosteroid induced osteoporosis and the increase in risk of fracture is one of the major problems. Great progress in the field of epidemiology, diagnosis, physiopathology and treatment has led to the elaboration of rational and efficient care of patients receiving corticosteroids. PMID- 15523294 TI - Effect of nesiritide (human b-type natriuretic peptide) and dobutamine on heart rate variability in decompensated heart failure. AB - BACKGROUND: Previous studies have suggested that natriuretic peptides may have direct sympathoinhibitory effects. Nesiritide (recombinant human B-type natriuretic peptide) has been recently approved for treatment of decompensated congestive heart failure (CHF). We sought to assess the effects of nesiritide compared with dobutamine on time-domain indices of heart rate variability (HRV) in patients with decompensated CHF. METHODS: The study population consisted of 185 patients, who were randomized to intravenous nesiritide at a low (0.015 microg/kg/min, n = 56) or high (0.03 microg/kg/min, n = 58) dose, or to dobutamine (> or = 5 microg/kg/min, n = 58). Time-domain HRV indices were obtained from 24-hour Holter recordings immediately before and during study drug therapy. RESULTS: Dobutamine therapy resulted in a decrease in standard deviation of the R-R intervals over a 24-hour period (SDNN), standard deviation of all 5 minute mean R-R intervals (SDANN), and the percentage of R-R intervals with >50 ms variation (pNN50) (all P < .05). Low-dose nesiritide induced an increase in SDNN (P < .05), and high-dose nesiritide resulted in a nonsignificant decrease in all measures of HRV. A significant interaction was noted between baseline HRV and the effect of vasoactive therapy on HRV (P = .028). Therefore, the effect of nesiritide and dobutamine was analyzed in relation to baseline HRV. In the dobutamine group, patients with moderately depressed HRV at baseline displayed a reduction in SDNN (P = .01), SDANN (P = .01), pNN50 (P = .04), and the square root of mean squared differences of successive R-R intervals (RMSSD) (P = .05), whereas no significant changes occurred in patients with severely depressed HRV. In the low-dose nesiritide group, patients with severely depressed HRV displayed an increase in SDNN (P = .001), SDANN (P = .02), and RMSSD (P = .01), with no significant changes in patients with moderately depressed HRV. HRV response to high-dose nesiritide was similar to that of dobutamine. CONCLUSIONS: Low-dose nesiritide therapy in patients with decompensated CHF improves indices of overall HRV and parasympathetic modulation, particularly if HRV is severely depressed at baseline. Dobutamine and possibly high-dose nesiritide can potentially lead to further deterioration of autonomic dysregulation. PMID- 15523295 TI - Left atrial thrombi, transient ischemic attacks, and atrial fibrillation. PMID- 15523296 TI - Regarding: Right ventricular cardiac dysfunction in HIV-infected patients studied with radionuclide ventriculography. PMID- 15523297 TI - Use of magnetic resonance imaging (MRI) to evaluate early ventricular mass reduction after aortic replacement. PMID- 15523298 TI - Sex-based selection of a method to relieve obstruction in hypertrophic cardiomyopathy: a hypothesis or more? PMID- 15523299 TI - Regarding: Statin use is associated with enhanced collateralization of severely diseased coronary arteries. PMID- 15523300 TI - Effect of obesity on risks after coronary artery bypass grafting. PMID- 15523301 TI - Evidence-based decision limits for cardiac troponin: low-level elevation and prognosis. PMID- 15523302 TI - Expanding indications for natriuretic peptides: importance of better new (research) protocols. PMID- 15523303 TI - Hypertension mega-trials with cardiovascular end points: effect of angiotensin converting enzyme inhibitors and angiotensin receptor blockers. AB - BACKGROUND: The renin-angiotensin-aldosterone system has a pivotal role in the short- and long-term regulation of blood pressure through its principal mediator, angiotensin II. Angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor blockers (ARBs) decrease the deleterious effects of angiotensin II on the vasculature and heart, but have different mechanisms of action. Although the blood pressure-lowering effect of ACE inhibitors and ARBs is equivalent to that of most other antihypertensive agents, emerging data suggest that these drug classes may have a greater effect on decreasing cardiovascular morbidity and mortality rates in specific patient populations. METHODS: We reviewed large (approximately > or =5000 patients) hypertension clinical trials using ACE inhibitors and ARBs and with cardiovascular morbidity/mortality end points. RESULTS: Six trials of ACE inhibitors and 5 trials of ARBs (3 completed, 2 ongoing) were selected for this analysis. Data from these hypertension mega trials suggest that ACE inhibitors and ARBs may decrease cardiovascular morbidity and mortality rates, especially in patients with diabetes mellitus, renal dysfunction, and left ventricular hypertrophy. However, some trials showed important blood-pressure differences and are therefore partly inconclusive for particular drug effects. CONCLUSIONS: Analysis of recently reported and ongoing mega-trials of renin-angiotensin-aldosterone system inhibitors may support the notion that their vasculoprotective properties confer greater benefit by virtue of their effects beyond blood-pressure reduction. Results from trials that will be completed in the next few years may provide further support of blocking the renin-angiotensin-system in cardiovascular protection in the management of hypertension. PMID- 15523304 TI - Magnetic resonance imaging of the peripheral vasculature. AB - BACKGROUND: Atherosclerotic disease of the peripheral vasculature is a prevalent condition for which catheter-based techniques have been considered to be the gold standard for diagnosis. However, because of their invasive nature, these techniques inherently have the potential for complications. Non-invasive diagnostic techniques have historically been limited by low accuracy and high operator dependence. Magnetic resonance angiography (MRA) is a new approach that has diagnostic accuracy comparable with invasive angiography. METHODS: The literature on MRA for evaluation of carotid, mesenteric, renal, and lower extremity arterial disease was extensively reviewed. Helpful diagnostic algorithms on the basis of the literature are also provided. RESULTS: MRA is both sensitive and specific when compared with invasive angiography for the evaluation of peripheral arterial disease and avoids the potential for complications resulting from arterial puncture and use of iodinated contrast. CONCLUSION: Current MRA techniques are diagnostically robust and have proven to be a highly accurate, safe, and convenient means of diagnosing atherosclerotic disease of the peripheral vascular system. PMID- 15523305 TI - Acute Catheterization and Urgent Intervention Triage strategY (ACUITY) trial: study design and rationale. AB - BACKGROUND: Patients with acute coronary syndromes (ACS; unstable angina and non ST-segment elevation myocardial infarction) are at significant risk for death and myocardial infarction. Early angiography followed by revascularization is considered the treatment of choice for moderate- to high-risk patients with ACS. However, despite the integration of newer therapies including stents, glycoprotein IIb/IIIa inhibitors, and thienopyridines, the rate of adverse ischemic events still remains unacceptably high, and the intensive pharmacologic regimens used to stabilize the disrupted atherosclerotic plaque and support angioplasty and surgical revascularization procedures elicit a high rate of bleeding complications. Pilot trials suggest that the thrombin-specific anticoagulant bivalirudin may improve clinical outcomes in ACS. STUDY DESIGN: In the Acute Catheterization and Urgent Intervention Triage strategY (ACUITY) trial, 13,800 patients with moderate- to high-risk ACS are being prospectively randomly assigned at up to 600 centers to unfractionated heparin or enoxaparin + IIb/IIIa inhibition, versus bivalirudin + IIb/IIIa inhibition, versus bivalirudin + provisional IIb/IIIa inhibition. All patients undergo cardiac catheterization within 72 hours, followed by percutaneous or surgical revascularization when appropriate. In a second random assignment, patients assigned to receive IIb/IIIa inhibitors are subrandomized to upstream drug initiation, versus IIb/IIIa inhibitor administration during angioplasty only. The primary study end point is the composite of death, myocardial infarction, unplanned revascularization for ischemia, and major bleeding at 30 days. Clinical follow-up will continue for 1 year. CONCLUSIONS: The ACUITY trial is the largest study yet performed in patients with ACS undergoing an invasive strategy. In addition to evaluating the utility of bivalirudin in ACS, this study will also provide important guidance regarding the necessity for and timing of IIb/IIIa inhibitor administration. PMID- 15523306 TI - Prognostic value of low-level cardiac troponin-I elevations in patients without definite acute coronary syndromes. AB - BACKGROUND: Low-level cardiac troponin-I (cTn-I) elevations predict adverse cardiovascular outcomes in patients with definite acute coronary syndromes (ACS), as defined by the presence of chest pain accompanied by ischemic electrocardiographic changes. However, their prognostic value in other clinical situations remains unclear. METHODS: We studied 366 patients with suspected myocardial infarction (MI) but without definite ACS, including 57 patients with low-level cTn-I elevations (1.0 to 3.0 ng/mL) and 309 patients with cTn-I <1.0 ng/mL. All cTn-I measurements were made with the Dade Stratus II analyzer. We determined the adjusted 1-year risk of nonfatal MI or death from coronary heart disease (CHD death) in each group by using Cox proportional hazards models. RESULTS: Among patients with cTn-I elevations between 1.0 and 3.0 ng/mL, 6 (11%) had a nonfatal MI or CHD death at 1 year compared with 12 (4%) patients in the cTn-I <1.0 ng/mL group [hazard ratio (HR), 3.5; 95% CI, 1.4 to 8.8]. After adjusting for baseline clinical characteristics, cTn-I levels between 1.0 and 3.0 ng/mL remained strongly associated with nonfatal MI or CHD death (adjusted HR, 3.4; 95% CI, 1.3 to 9.4). This association persisted even in the 215 patients who presented without chest pain (adjusted HR, 4.3; 95% CI, 1.4 to 13). CONCLUSIONS: Low-level cTn-I elevations identify a subset of patients at increased risk for future cardiovascular events, even when obtained outside the context of definite ACS or presentation with chest pain. PMID- 15523307 TI - Detection of increased temperature of the culprit lesion after recent myocardial infarction: the favorable effect of statins. AB - BACKGROUND: Increased thermal heterogeneity has been demonstrated in atherosclerotic plaques, with the higher temperature recorded in acute myocardial infarction (MI). Dietary or treatment interventions reduce heat production. The purpose of the present study was to investigate whether increased plaque temperature is maintained for a prolonged period after MI and the role of statin administration. METHODS: We enrolled 55 patients, 29 with recent MI and 26 with chronic stable angina (CSA). Total cholesterol, C-reactive protein (CRP), interleukin-6 (IL-6) and soluble adhesion molecules were measured in the study population. All patients underwent coronary plaque temperature measurements. Temperature difference (DeltaT) was designated as the temperature of the culprit atherosclerotic plaque minus the temperature of the proximal healthy vessel wall. RESULTS: Under treatment with statins were 19 patients with recent MI and 14 with CSA. In patients with recent MI DeltaT was 0.19 +/- 0.18 degrees C, while in patients with CSA was 0.10 +/- 0.08 degrees C (P = .03). Patients treated with statins had lower DeltaT compared to untreated patients (0.10 +/- 0.11 versus 0.20 +/- 0.18 degrees C, P = .01). Treated patients with recent MI had similar DeltaT compared to CSA patients treated with statins (0.13 +/- 0.13 versus 0.07 +/- .06 degrees C, P = .14), while untreated patients with recent MI had substantially increased DeltaT compared to untreated patients with CSA (0.28 +/- 0.22 versus 0.14 +/- 0.10 degrees C, P = .04). DeltaT was positively correlated with CRP (R = 0.50, P < .01), IL-6 (R = 0.58, P < .01), and intercellular adhesion molecule-1 (R = 0.40, P = .03) levels. CONCLUSION: Increased plaque temperature is observed for an extended period after myocardial infarction, indicating that the inflammatory process is sustained after plaque rupture. Statins have a beneficial effect after MI on plaque temperature. PMID- 15523308 TI - Impaired coronary blood flow in patients with metabolic syndrome: documented by Thrombolysis in Myocardial Infarction (TIMI) frame count method. AB - BACKGROUND: Endothelium plays an important role in regulating coronary vascular tone. In addition, several of cardiovascular risk factors that are associated metabolic syndrome have been reported to be associated with endothelial dysfunction. In the present study we aimed to evaluate the coronary blood flow in patients with metabolic syndrome by means of the Thrombolysis in Myocardial Infarction (TIMI) frame count. METHOD: Forty-two patients with metabolic syndrome (group I) and 42 control subjects without metabolic syndrome (group II) were included in the study. All subjects had angiographically proven normal coronary arteries. Diagnosis of metabolic syndrome was based on the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) guidelines published in 2001. Coronary flow rates of all subjects were documented by TIMI frame count method. RESULTS: TIMI frame counts for each of the major epicardial coronary arteries were found to be significantly higher in patients with metabolic syndrome compared with control subjects (corrected TIMI frame count for left anterior descending coronary artery: 35 +/- 7 vs 25 +/- 7, respectively; left circumflex coronary artery: 32 +/- 9 vs 25 +/- 7, respectively; right coronary artery: 31 +/- 9 vs 24 +/- 5, respectively; P < .001 for all). Statistically significant independent relationships were found between TIMI frame count and body mass index (R(2) = 0.480, P = .009), waist circumference (R2 = 0.551, P = .001), and triglyceride level (R2 = 0.434, P = .036). CONCLUSION: We have shown for the first time that patients with metabolic syndrome and angiographically normal coronary arteries have higher TIMI frame counts for all 3 coronary vessels, indicating impaired coronary blood flow, compared to control subjects without metabolic syndrome. PMID- 15523309 TI - Prognostic value of predischarge electrocardiographic measurement of infarct size after thrombolysis: insights from GUSTO I Economics and Quality of Life substudy. AB - BACKGROUND: Current methods for risk stratification after acute myocardial infarction (MI) include several noninvasive studies. In this cost-containment era, the development of low-cost means should be encouraged. We assessed the ability of an electrocardiogram (ECG) MI-sizing score to predict outcomes in patients enrolled in the Economics and Quality of Life (EQOL) sub study of the Global Utilization of Streptokinase and Tissue plasminogen activator for Occluded coronary arteries -I (GUSTO-I) trial. METHODS: We classified patients by electrocardiographic Selvester QRS score at hospital discharge: those with a score 0-9 versus > or =10. Endpoints were 30-day and 1-year mortality, resource use, and quality-of-life measures. RESULTS: Patients with a QRS score <10 were well-matched with those with QRS score > or =10 with the exception of a trend to more anterior MI in the higher scored group. Patients with QRS score > or =10 had increased risk of death at 30-days (8.9% vs. 2.9% P < .001), and this difference persisted at 1 year (12.6% vs. 5.4%, P = .001). Recurrent chest pain, use of angiography, and angioplasty were similar during follow-up. However, there was a trend toward less coronary bypass surgery in patients with a QRS score > or =10. Readmission rates were higher at 30 days but similar at 1 year. CONCLUSIONS: Stratification of patients after acute MI by a simple measure of MI size identifies populations with different long-term prognoses; patients with a QRS score > or =10 (approximately 30% of the left ventricle infarcted) at discharge have poorer outcomes in both the short- and long-term. The standard 12-lead ECG provides a simple, economical means of risk stratification at discharge. PMID- 15523310 TI - Alcohol intake is not associated with subclinical coronary atherosclerosis. AB - BACKGROUND: The inverse relation between alcohol intake and clinical coronary artery disease (CAD) is well established, although the mechanisms remain speculative. We studied the relation between alcohol intake and subclinical CAD to assess the possible role of alcohol in atherogenesis. METHODS: We conducted a prospective study of 731 consecutive, consenting, active-duty US Army personnel (39 to 45 years of age) without known CAD who were undergoing a routine physical examination. Each participant was surveyed with the validated Block dietary questionnaire, which included detailed information on alcohol intake as wine, beer, or liquor. Subclinical CAD was determined by means of electron beam computed tomography to quantify coronary artery calcification (CAC). RESULTS: The mean age was 42 (+/-2); 83% were male, 71% were white, and 82% were college graduates. The prevalence of CAC was 18.6% (mean CAC score = 12 +/- 69). Twenty two percent drank alcohol daily, with an average of 2.4 drinks per day. Systolic blood pressure was correlated with number of drinks per day (r = 0.10, P = .025). Among drinkers, HDL was weakly correlated with daily alcohol consumption (r = 0.10, P = .025). There was no relation between the CAC score and the alcohol intake as measured by drinks per day (OR, 1.02; 95% CI, 0.64 to 1.63; 1.13, 0.59 to 2.15; 1.26, 0.69 to 2.59, for less than 1, 1 to 2, and more than 2 drinks per day, respectively). Stratified analyses based on type of alcohol and multivariate analyses indicated no independent relation between any type or quantity of alcohol intake and the presence or extent of coronary calcification. CONCLUSIONS: Alcohol intake does not appear to be inversely related to subclinical CAC, implying that previous observations of a protective effect of alcohol on clinical CAD may involve factors related to plaque stability rather than atherogenesis. PMID- 15523311 TI - Electrocardiographic findings in cardiogenic shock, risk prediction, and the effects of emergency revascularization: results from the SHOCK trial. AB - OBJECTIVES: To evaluate electrocardiographic (ECG) parameters as predictors of 1 year mortality in patients developing cardiogenic shock after acute myocardial infarction (AMI), and to document associations between these ECG parameters and the survival benefit of emergency revascularization versus initial medical stabilization. BACKGROUND: Emergency revascularization reduces the risk of mortality in patients developing cardiogenic shock after AMI. The prognostic value of ECG parameters in such patients is unclear, and it is uncertain whether emergency revascularization reduces the mortality risk denoted by ECG parameters. METHODS: In a prospective substudy of 198 SHOCK (SHould we emergently revascularize Occluded Coronaries for cardiogenic shocK) trial patients, ECGs recorded within 12 hours of shock were interpreted by personnel blinded to the patients' treatment assignment and outcome. RESULTS: The baseline heart rate was higher in non-survivors than in survivors (106 +/- 20 versus 95 +/- 24 beats/minute, P = .001). There was a significant association between the QRS duration and 1-year mortality in medically stabilized patients (115 +/- 28 ms in non-survivors versus 99 +/- 23 ms in survivors, P = .012), but not in emergently revascularized patients (110 +/- 31 versus 116 +/- 27 ms respectively, P = .343). The interaction between the QRS duration, mortality and treatment assignment was significant (P = .009). Among patients with inferior AMI, a greater sum of ST depression was associated with higher 1-year mortality in medically stabilized patients (P = .029), but not in emergently revascularized patients (P = .613, treatment interaction P = .025). On multivariate analysis, the independent mortality predictors were increasing age, elevated pulmonary capillary wedge pressure, heart rate, sum of ST depression in medically stabilized patients, and interaction (P = .016) between a prolonged QRS duration and treatment assignment. The adjusted hazard ratio for 1-year mortality per 20 ms increase in the QRS duration was 1.19 (95% CI 0.98-1.46) in medically stabilized patients and 0.81 (95% CI 0.63-1.03) in emergently revascularized patients. CONCLUSION: ECG parameters identified patients with cardiogenic shock who were at high risk. Emergency revascularization eliminated the incremental mortality risk associated with cardiogenic shock in patients with a prolonged QRS duration, or inferior AMI accompanied by precordial ST depression. Prospective assessments of the magnitude of the treatment effect based on ECG parameters are required. PMID- 15523312 TI - Oxidized low-density lipoprotein levels circulating in plasma and deposited in the tissues: comparison between Helicobacter pylori-associated gastritis and acute myocardial infarction. AB - BACKGROUND: Oxidized low-density lipoprotein (ox-LDL) is a key factor in the progression of atherosclerosis. We developed a sensitive method for measuring plasma ox-LDL levels using a novel anti-ox-LDL antibody. Recently, several studies have shown positive associations between Helicobacter pylori (H pylori) infection and coronary heart disease. Thus the question arises whether an increase in the plasma levels of ox-LDL occurs in patients with H pylori gastritis. METHODS: We measured plasma ox-LDL levels in patients with H pylori gastritis (n = 27) and compared them with those in patients with acute myocardial infarction (AMI) (n = 62) and stable angina pectoris (SAP; n = 63) and those in control subjects (n = 64). In addition, ox-LDL localization and the presence of macrophages and neutrophils were studied immunohistochemically in gastritis specimens and in coronary culprit lesions obtained from patients with AMI. RESULTS: Plasma ox-LDL levels in patients with AMI were significantly higher than those in patients with SAP (P <.0001), patients with H pylori gastritis (P <.0001), or in control subjects (P <.0001; AMI, 1.34 +/- 0.95; SAP, 0.61 +/- 0.29; Gastritis, 0.53 +/- 0.17; control, 0.57 +/- 0.23 ng/5 microg LDL protein). Immunohistochemically, H pylori gastritis specimens showed distinct infiltration of macrophages and myeloperoxidase-positive neutrophils; however, ox-LDL localization was not detected. In contrast, coronary culprit plaques revealed strong positivity for ox-LDL in ruptured lipid cores with abundant macrophage derived foam cells, and these plaques also contained myeloperoxidase-positive neutrophils. CONCLUSION: Our results suggest that plasma ox-LDL levels do not seem to be associated with H pylori infection, but do relate to coronary plaque instability in AMI. PMID- 15523313 TI - High-grade infarct-related stenosis after successful thrombolysis: strong predictor of reocclusion, but not of clinical reinfarction. AB - BACKGROUND: After successful thrombolysis, a high-grade stenosis at 24-hour angiography is strongly predictive of reocclusion and is often believed to result in high reinfarction rates. However, routine angioplasty did not reduce death or reinfarction in past trials. Systematic angiographic follow-up shows that reocclusion often occurs without clinical reinfarction. This study investigates whether the increased risk for reocclusion associated with a high-grade lesion translates into impaired clinical outcome. METHODS: In the ischemia-guided Antithrombotics in the Prevention of Reocclusion in COronary Thrombolysis (APRICOT-1) trial, 240 patients with ST-elevation MI who had an open infarct artery 24 hours after thrombolysis had 3-month repeat angiography to assess reocclusion, with clinical follow-up at 3 months and 3 years. RESULTS: On the basis of the optimal discriminative stenosis severity, the reocclusion rate was 40% (47/118) in patients with a high-grade residual stenosis and 16% (20/122) in patients with a low-medium-grade lesion (risk ratio [RR], 2.43; 95% CI, 1.54 3.84; P <.01). Three-month death and reinfarction rates did not differ: 6% (7/118) versus 9% (11/122; RR, 0.66; 95% CI, 0.26-1.64; P = not significant). Systematic angiographic follow-up revealed that reocclusion of a high-grade lesion occurred in the absence of clinical reinfarction in 85% (40/47) of patients, as compared with 45% (9/20) in patients with a low-medium-grade stenosis (RR, 1.89; 95% CI, 1.15-3.12; P <.01). Despite an independent association with reocclusion, a high-grade stenosis was not predictive of either short- or long-term death and reinfarction. CONCLUSIONS: After successful thrombolysis and adopting an ischemia-guided revascularization strategy, patients with a high-grade stenosis experience death/reinfarction rates similar to that of patients with a low-medium-grade lesion. This is true despite a 2- to 3-fold higher risk for reocclusion. The finding that reocclusion of a high-grade lesion often occurs without clinical reinfarction explains the absence of a relationship between a severe stenosis and death/reinfarction. Appreciation of these observations may contribute to an optimal design of a future randomized trial to re-evaluate the impact of a routine invasive strategy. PMID- 15523314 TI - Effect of age on the use of evidence-based therapies for acute myocardial infarction. AB - BACKGROUND: Previous studies have documented an underuse of evidence-based therapies in patients with acute myocardial infarction (AMI). However, many of these studies failed to consider contraindications to therapy, the effect of age (ie, elderly vs non-elderly patients) on use, or both. The objective of this study was to determine whether elderly patients are less likely than non-elderly patients to receive evidence-based AMI treatments, both before and after the consideration of contraindications to therapy. METHODS: A retrospective chart review of a random sample of 5131 patients with AMI who were admitted to 1 of 44 hospitals in Ontario was conducted for the fiscal years 1994 to 1996. Using the Canadian Cardiovascular Research Team (CCORT)/Canadian Cardiovascular Society (CCS) Quality Indicators for AMI Care, we classified patients as being eligible or ideal (ie, no contraindications to treatment) candidates to receive aspirin, beta-blockers, thrombolysis, angiotensin-converting enzyme inhibitors (ACEIs), or statins or to undergo lipid profiling. The proportions of eligible and ideal patients who received treatment were calculated, and the latter were compared with benchmarks. RESULTS: The median age of the cohort was 69 years; 63% were of the patients were aged > or =65 years. There was underperformance of prescribing treatments in ideal candidates relative to benchmarks (eg, aspirin at discharge: 78.6% vs 90% benchmark). The odds of ideal (ie, no contraindications) elderly candidates receiving various evidence-based AMI treatments were consistently less than that of non-elderly patients with AMI, with the exception of ACEIs at discharge (odds ratio, 1.46; 95% CI, 1.22-1.74). CONCLUSIONS: Despite adjustments for contraindications to therapy, the underuse of AMI treatments, particularly in elderly patients, was found. PMID- 15523315 TI - Color of culprit lesion at 6 months after plain old balloon angioplasty versus stenting in patients with acute myocardial infarction. AB - BACKGROUND: Although the healing process of disrupted yellow plaques at myocardial infarction (MI) culprit lesions has been reported, the effect of stenting on this process has not been clarified. Stenting has been reported to deteriorate the endothelial function after percutaneous coronary intervention (PCI). Therefore, we compared the angioscopic morphology of culprit lesions at 6 months after plain old balloon angioplasty (POBA) and stenting to clarify the effect of stenting on the healing of disrupted culprit plaques of acute MI. METHODS: Patients with acute MI who had yellow culprit plaque, successful reperfusion therapy with POBA (n = 21) or stenting (n = 22), and a successful 6 month follow-up angioscopic examination were included in this study. Oral ticlopidine (200 mg/day) was administered for 3 to 6 months after stenting. RESULTS: At 6 months after reperfusion therapy, the color of the culprit lesion became white in significantly more patients treated with stenting than treated with POBA (50% vs 14%; P = .01). However, the prevalence of thrombus appeared to be higher in patients treated with stenting than in patients treated with POBA (27% vs 5%; P = .04). Although there was some difference in the patients' characteristics in the groups, logistic regression analysis revealed no significant influence of those factors on the color of or on the prevalence of thrombus at the culprit lesion. CONCLUSIONS: Coronary stenting in patients with acute MI leads to the disappearance of yellow color at a significantly higher rate than POBA; however, whether it stabilizes the plaque requires further investigation. PMID- 15523316 TI - Endothelial nitric oxide synthase gene polymorphism and ischemic heart disease. AB - BACKGROUND: Ischemic heart disease (IHD), the leading cause of death worldwide, is a multifactorial disease arising from the complex interplay of genetic and environmental factors. There is considerable evidence that nitric oxide (NO) plays an important role in protecting against atherosclerosis, the process underlying IHD. Endothelial NO is synthesized from the amino acid l-arginine by the endothelial isoform of NO synthase (eNOS). Thus, polymorphisms of the eNOS gene, by altering production of NO within the vascular endothelium, are potential risk factors for IHD. Several groups have investigated the role of the G894T polymorphism of the eNOS gene in IHD by using case-control association studies; however, its role is unclear because of contradictory results from these studies. We applied family-based association tests to investigate the role of this polymorphism in IHD in a well-defined Irish population. METHODS: A total of 1023 individuals from 388 families (discordant sibships and parent-offspring trios) were recruited. Linkage disequilibrium between the polymorphism and IHD was tested for by using the combined transmission disequilibrium test/sib transmission disequilibrium test and pedigree disequilibrium test. RESULTS: Both the combined transmission disequilibrium test/sib-transmission disequilibrium test and pedigree disequilibrium test analyses found no statistically significant excess transmission of either allele to affected individuals (P = .57 and P = .38, respectively). CONCLUSIONS: Using robust family-based association tests specifically designed for the study of complex diseases, we found no evidence that the G894T polymorphism of the eNOS gene has a significant role in the development of IHD in our population. PMID- 15523317 TI - Sex-related differences in outcome after ST-segment elevation myocardial infarction treated by primary angioplasty: data from the Zwolle Myocardial Infarction study. AB - BACKGROUND: Several studies have found that among patients with ST-elevation myocardial infarction (STEMI) treated by thrombolysis, female sex is associated with a worse outcome. The aim of this study was to investigate sex-related differences in clinical and angiographic findings and in long-term outcome in patients with STEMI treated with primary angioplasty. METHODS: Our population is represented by 1548 consecutive patients with STEMI treated by primary angioplasty from April 1997 to October 2001. All clinical, angiographic, and follow-up data were prospectively collected. RESULTS: Among 1548 patients, 353 were women (22.8%). Female sex was associated with more advanced age, higher prevalence of diabetes, hypertension, more advanced Killip class, longer ischemia time, and smaller vessel caliber. No difference was observed in terms of procedural success, postprocedural epicardial flow, myocardial perfusion, ST segment resolution, and enzymatic infarct size. At 1-year follow-up, female sex was associated with a significantly higher 1-year mortality rate at univariate (9.3% vs 4.9 %, RR [95% CI] = 1.79 [1.14 to 2.8], P = .002) but not at multivariate analysis (RR [95% CI] = 1.41 [0.86 to 2.32], P = NS). CONCLUSIONS: This study shows that in patients with STEMI treated by primary angioplasty, women are associated with higher mortality rate in comparison with men, mainly because of their high-risk profile and angiographic features. Female sex did not emerge as an independent predictor of death. PMID- 15523318 TI - Rapamycin-eluting stents for the treatment of bifurcated coronary lesions: a randomized comparison of a simple versus complex strategy. AB - BACKGROUND: Rapamycin-eluting stents (RES) have been shown to reduce restenosis in many types of lesions. However, the ideal strategy for the treatment of coronary bifurcated lesions has not been established to date. This randomized study compares 2 strategies for the RES treatment of bifurcation lesions: a simple approach (stenting the main vessel and balloon dilatation for the side branch [SB]) versus a complex approach (stents for both vessels). METHODS: To compare both strategies, a randomized study was conducted in 91 patients with true coronary bifurcation lesions. All patients received an RES at the main vessel, covering the SB. Patients from group A (n = 47) were assigned to balloon dilation of the involved SB (simple strategy); patients in group B (n = 44) were randomized to receive a second stent at the SB origin (complex strategy). There were no differences between groups regarding baseline clinical and angiographic data. RESULTS: Major adverse cardiac events occurred in 3 patients from group A (2 non-Q-wave myocardial infarctions and 1 target lesion revascularization). Six month angiographic reevaluation was obtained in 80 patients (88%). Restenosis of the main vessel was observed in 1 (2%) patient from group A and in 4 (10%) from group B. Restenosis of the SB appeared in 2 (5%) patients from group A and in 6 (15%) from group B. CONCLUSIONS: Both strategies are effective in reducing the restenosis rate, with no differences in terms of clinical outcome. Elective SB stenting seems to provide no advantages over the simpler stent jail followed by SB balloon dilation. PMID- 15523319 TI - Regression of significant tricuspid regurgitation after mitral balloon valvotomy for severe mitral stenosis. AB - BACKGROUND: Significant tricuspid regurgitation (TR) is occasionally associated with severe mitral stenosis and has an adverse impact on morbidity and mortality in patients undergoing mitral valve surgery. However, the effect of successful mitral balloon valvotomy (MBV) on significant TR is not fully elucidated. The aim of this study was to investigate TR after MBV in patients with severe mitral stenosis. METHODS: We analyzed the data of 53 patients with significant TR (grade > or =2, on a 1 to 3 scale) from the mitral balloon valvotomy database at our hospital. Patients were evaluated by Doppler echocardiography before valvotomy and at follow-up 1 to 13 years after MBV. Patients were divided into group A (27 patients), in whom TR regressed by > or =1 scale, and group B (26 patients), in whom TR did not regress. RESULTS: The Doppler-determined pulmonary artery systolic pressure was initially higher and decreased at follow-up more in group A (from 70.7 +/- 23.8 to 36.5 +/- 8.3 mm Hg; P < .0001) than in group B (from 48.7 +/- 17.8 to 41.6 +/- 13.1 mm Hg; P = NS). Compared with patients in group B, patients in group A were younger (25 +/-10 vs 35 +/- 11 years; P < .005), had higher prevalence of functional TR (85% vs 8%; P < .0001), and had lower incidence of atrial fibrillation (7% vs 38%; P < .005). Significant decrease in right ventricular end-diastolic dimension after MBV was noted in group A but not in group B. The mitral valve area at late follow-up was larger in group A than in group B (1.8 +/- 0.3 vs 1.6 +/- 0.3 cm2; P < .05). CONCLUSIONS: Regression of significant TR after successful MBV in patients with severe mitral stenosis was observed in patients who had severe pulmonary hypertension. This improvement in TR occurred even in the presence of organic tricuspid valve disease. PMID- 15523320 TI - Underlying risk factors incrementally add to the standard risk estimate in detecting subclinical atherosclerosis in low- and intermediate-risk middle-aged asymptomatic individuals. AB - BACKGROUND: Traditional risk factors predict the 10-year risk of developing coronary heart disease (CHD). Underlying risk factors like physical inactivity, obesity (BMI >30 kg/m2), and family history of premature CHD are independently associated with CHD. High burden of coronary artery calcification (CAC) on electron beam tomography (EBT) is a CHD risk equivalent. GOALS: To determine the association between CAC and traditional risk assessment and whether the presence of added underlying risk factors is associated with advanced CAC in low- and intermediate-risk individuals. METHODS: After excluding patients with CHD, we studied 8549 asymptomatic individuals referred for EBT for cardiac risk assessment. Traditional myocardial infarction risk score was estimated according to Framingham criteria, and individuals were divided into 3 groups: low-risk (< or =9% MI risk over the next 10 years), intermediate-risk (10%-20% risk), and high-risk (>20 % risk). Advanced CAC was defined as a calcium score > or =75th percentile based on sex and age. RESULTS: The prevalence of advanced CAC was 20% in low-risk, 27% in intermediate-risk, and 31% in high-risk individuals (P < .001). Underlying risk factors were incrementally associated with advanced CAC in low- and intermediate-risk individuals (P < .001). A receiver operating characteristic curve analysis revealed that addition of underlying risk factors to traditional risk assessment increased the area under the curve significantly from 0.56 to 0.62 (P < .001). CONCLUSION: A substantial proportion of low- and intermediate-risk individuals have advanced CAC. Adding family history, obesity, and physical inactivity to traditional risk assessment improves prediction of advanced CAC, which may lead these individuals to be treated more aggressively at an earlier age. PMID- 15523321 TI - Impact of short-term intermittent intravenous dobutamine therapy on endothelial function in patients with severe chronic heart failure. AB - BACKGROUND: Intermittent intravenous dobutamine therapy is used to treat patients with decompensated end-stage chronic heart failure (CHF), in whom the vascular endothelium is usually impaired. Although it has been suggested that modification or reversal of endothelial dysfunction may be of significant therapeutic benefit, the impact of short-term intermittent intravenous dobutamine therapy on flow mediated dilation (FMD) in patients with severe decompensated end-stage chronic CHF has not been assessed. METHODS: We prospectively assessed the impact of intermittent intravenous low-dose dobutamine therapy on endothelium-dependent brachial artery FMD and endothelium-independent nitroglycerin (NTG)-mediated vasodilation using high resolution ultrasound scanning in 20 consecutive male patients with severe CHF and ischemic cardiomyopathy (New York Heart Association functional class IV), at baseline and after 4 months, and compared them to 20 age and sex-matched control subjects. The cardiac index (CI), stroke index (SI), and systemic vascular resistance (SVR) were assessed non-invasively with a thoracic electrical bioimpedance device before and after intravenous dobutamine therapy. RESULTS: Intermittent intravenous dobutamine therapy resulted in significant improvement in post-intervention FMD compared with baseline (7.7% +/- 2.4% vs 1.1% +/- 2.6%; P = .001), a finding not evident in control subjects (1.3% +/- 2.6% vs 1.2% +/- 2.1%; P = .78). There was no significant effect of dobutamine treatment compared with control subjects on NTG-induced vasodilation (7.6% +/- 5.5% vs 7.5% +/- 8.8%, P = .979). Short-term dobutamine therapy also significantly improved SVR (1797 +/- 926 dyne sec/cm5 vs 2172 +/- 1133 dyne sec/cm5, P = .05), CI (2. 4+/- 0.6 L/min/m2 vs 1.9 +/- 0.6 L/min/m2, P = .01), and SI (33.5 +/- 11.7 mL/m(2) vs 27.2 +/- 12.4 mL/m2, P = .02). CONCLUSIONS: Short-term intermittent intravenous low-dose dobutamine therapy significantly improved vascular endothelial function, perhaps demonstrating an additional mechanism for improved SVR, CI, and SI in patients with severe CHF. PMID- 15523322 TI - Effects of diabetes mellitus and ischemic heart disease on the progression from asymptomatic left ventricular dysfunction to symptomatic heart failure: a retrospective analysis from the Studies of Left Ventricular Dysfunction (SOLVD) Prevention trial. AB - BACKGROUND: Emerging data suggest that diabetes mellitus is a risk factor for the progression of established heart failure only in those patients with ischemic cardiomyopathy. Whether diabetes mellitus is a risk factor for the progression from asymptomatic left ventricular systolic dysfunction to symptomatic heart failure in patients with left ventricular dysfunction of an ischemic cause is not known. METHODS: We performed a retrospective analysis of 2821 patients with asymptomatic left ventricular systolic dysfunction from the Studies of Left Ventricular Dysfunction (SOLVD) Prevention trial. We used adjusted survival analysis to examine the effects of ischemic heart disease and diabetes mellitus on 3 prespecified study end points: (1) development of heart failure (HF) symptoms, (2) HF hospitalization, and (3) death or development of symptoms. RESULTS: There is a statistically significant interaction between the cause of left ventricular systolic dysfunction and diabetes mellitus on the risk of development of heart failure symptoms (P = .020). Patients with ischemic cardiomyopathy and diabetes had an increased risk of progression to symptomatic heart failure (HR = 1.56, P < .001), hospitalization for heart failure (HR = 2.16, P < .001), and death or development of symptoms (HR = 1.50, P < .001), compared with patients with ischemic cardiomyopathy without diabetes. In contrast, diabetes was not associated with an increased risk of reaching these end points in patients with nonischemic cardiomyopathy. CONCLUSIONS: Diabetes mellitus is a risk factor for the progression from asymptomatic left ventricular systolic dysfunction to symptomatic heart failure, but this risk appears to be confined to those patients with ischemic cardiomyopathy. PMID- 15523323 TI - Impact of angiotensin-converting enzyme gene polymorphism on neurohormonal responses to high- versus low-dose enalapril in advanced heart failure. AB - BACKGROUND: The impact of angiotensin-converting enzyme (ACE) gene polymorphism on neurohormonal dose response to ACE inhibitor therapy is unclear. METHODS: ACE Insertion (I) or Deletion (D) genotype was determined in 74 patients with chronic heart failure who were randomly assigned to receive either high-dose or low-dose enalapril over a period of 6 months. Monthly pre-enalapril and post-enalapril neurohormone levels (serum ACE activity (sACE), plasma angiotensin II (A-II), plasma renin activity (PRA), and serum aldosterone (ALDO) were compared between genotype subgroups and between patients who received high- or low-dose enalapril within each genotype subgroup. RESULTS: At baseline, predose/postdose sACE and postdose PRA were significantly higher in the DD genotype. At 6-month follow-up, postdose sACE was reduced in a dose-dependent fashion in all three genotypes (P < .05). However, predose and postdose ALDO and A-II levels did not differ between each genotype subgroup at baseline or by enalapril dose within each genotype subgroup. ALDO escape and A-II reactivation were not affected by ACE genotype or enalapril dosage. CONCLUSIONS: Predose sACE were consistently higher in the DD genotype when compared with ID or II subgroups. Despite a dose-dependent suppression of sACE, there were no observed statistically significant differences in ALDO and A-II suppression or escape with escalating doses of enalapril within each subgroup. PMID- 15523324 TI - Utility of B-natriuretic peptide in the evaluation of left ventricular diastolic function: comparison with tissue Doppler imaging recordings. AB - BACKGROUND: Although Doppler echocardiography provides assessment of abnormal left ventricular (LV) diastolic filling dynamics, its inherent limitations suggest the need for additional measures of diastolic dysfunction. The ratio of mitral velocity to early diastolic velocity of the mitral annulus (E/E') derived from tissue Doppler imaging is associated with the mean LV end diastolic pressure (LVEDP). Because data suggest that B-natriuretic peptide (BNP) reflects ventricular pressure, we hypothesized that BNP levels correlate with indices of LV diastolic function by tissue Doppler imaging. METHODS AND RESULTS: Doppler recordings of the mitral inflow and tissue Doppler imaging of the mitral annulus were obtained in 108 patients referred for echocardiography. BNP levels were measured by a rapid immunoassay and blinded to cardiologists making the assessment of LV function. BNP levels were higher when elevated E/E' ratios derived from tissue Doppler imaging were identified. Patients identified with elevated LVEDP, defined as E/E' >15 (n = 16), had the highest BNP levels (463 +/- 80 pg/mL). Those with normal LVEDP, as predicted by E/E' <8 (n = 36), had a mean BNP concentration of 97 +/- 27 pg/mL. Those with E/E' of 8 to 15 (n = 48) had a mean BNP level of 122 +/- 24 pg/mL. The area under the receiver-operating curve for BNP to detect E/E' >15 was 0.89 (95% CI, 0.82 to 0.96; P < .001). A BNP value of 173 pg/mL had a sensitivity of 88%, a specificity of 82%, and an accuracy of 83% for predicting E/E' >15. CONCLUSIONS: BNP levels correlate with the E/E' ratios from tissue Doppler imaging (r = 0.48) and can reliably provide estimation of LV filling pressures. Although BNP levels do not correlate well with E/E' ratios across the full spectrum of values, the combination of BNP levels along with E/E' ratios from tissue Doppler imaging may be a better predictor of elevated LV filling pressures in patients with suspected diastolic dysfunction. PMID- 15523325 TI - Systole-diastole mismatch in hypertrophic cardiomyopathy is caused by stress induced left ventricular outflow tract obstruction. AB - BACKGROUND: Pharmacological stress is used to assess the degree of left ventricular (LV) subvalvular gradient in patients with hypertrophic cardiomyopathy (HCM), but there is little information about associated physiological changes. METHODS: Echocardiography-Doppler ultrasound scanning measurements in 23 patients with HCM and 23 control subjects of similar age were studied at rest and at the end point of dobutamine stress. RESULTS: In patients, the systolic time was normal at rest, but increased abnormally with stress. In patients, the total isovolumic contraction time failed to shorten, and the total ejection time increased abnormally. Changes in total ejection time correlated with an increase in peak subvalvular gradient in control subjects and patients (r = 0.52 and r = 0.66, respectively; P <.01 for both). In patients, the diastolic time was normal at rest, but shortened abnormally with stress. In patients, the isovolumic relaxation time fell abnormally, as did the filling time. Mitral E wave acceleration and left atrium size were unchanged with stress in control subjects, but consistently increased in patients with HCM, which indicates an increased early diastolic atrioventricular pressure gradient. CONCLUSION: In HCM, systolic period increases abnormally with stress. This is not because of a loss of inotropy, but is directly related to the degree of LV outflow tract obstruction. As a result, the diastolic period fails to increase, reducing the time available for coronary flow, the LV filling pattern is modified, and the diastolic atrioventricular pressure gradient increases. These changes may contribute to symptom development and suggest why reducing LV outflow tract obstruction per se may be therapeutically useful in HCM. PMID- 15523326 TI - Predicting maximum heart rate among patients with coronary heart disease receiving beta-adrenergic blockade therapy. AB - BACKGROUND: The use of beta-adrenergic blockade (BB) therapy is common among patients with coronary heart disease (CHD), and as a result, these patients often undergo exercise testing while taking these medications. However, evaluation of maximal voluntary effort during exercise testing is often in question because current equations to predict maximum heart rate (HR(max); eg, 220 - age) are based on subjects without heart disease or BB therapy. The purpose of this study was to develop and validate an age-specific equation to predict HR(max) in patients with CHD who are receiving BB therapy. METHODS: We queried the Henry Ford Preventive Cardiology Outcomes database for patients with a history of myocardial infarction or revascularization procedure; preserved left ventricular function; age, 40 to 80 years; sinus rhythm; and a graded treadmill test with a respiratory exchange ratio > or =1.10. Data were split, based on date, such that tests performed between November 1996 and April 2001 were used as the BB prediction equation development group (n = 334; 73% men) and those performed between May 2001 and April 2002 were used as the BB cross-validation group (n = 94; 84% men). Linear regression was used to develop the equation to predict HR(max), based on age, and to calculate the correlation coefficient of the prediction equation among the cross-validation group. RESULTS: The resultant prediction equation was HR(max) = 164 - 0.7 x age (r2 = 0.13), with a standard error of the estimate of 18 per minute. Among the cross-validation group, mean predicted HR(max) was not significantly different from mean measured HR(max) (P = .7). The mean error of prediction was -0.4 +/- 2.0 per minute (mean +/- SEM), and the correlation was r = 0.38. CONCLUSIONS: This new equation provides a better estimate of HR(max) for patients with CHD receiving BB therapy than previously reported equations. Additional variables may improve the equation but may not be as convenient to use. PMID- 15523327 TI - Association between chronic heart failure and inhaled beta-2-adrenoceptor agonists. AB - BACKGROUND: Recent reports suggest an association between beta-agonists and the risk of incident chronic heart failure (CHF). We sought to examine the association between inhaled beta-agonists and risk of incident and nonincident heart failure. METHODS: We performed a nested case-control study within the Ambulatory Care Quality Improvement Project (ACQUIP). Case subjects were defined as having had a hospitalization with a primary discharge diagnosis of CHF. Controls were randomly selected from the ACQUIP cohort. The exposure was the number of beta-agonist canisters filled in the 90 days before an index date. RESULTS: After adjusting for potentially confounding factors, there appeared to be no association between the use of inhaled beta-agonists and the risk of heart failure (1-2 canisters per month, OR 1.3 [95% CI 0.9, 1.8], > or =3 canisters per month, 1.1 [95% CI 0.8, 1.6]). However, among the cohort that had a history of CHF, there appeared to be a dose-response association between the number of inhaled beta-agonists and the risk of hospitalization for chronic heart failure (1-2 canisters per month, adjusted OR 1.8 [95% CI 1.1, 3.0], > or =3 canisters per month, adjusted OR 2.1 [95% CI 1.2, 3.8]). CONCLUSION: beta-Agonists did not appear to be associated with incident heart failure but were associated with risk of CHF hospitalization among those subjects with a previous CHF diagnosis. Although a causal relationship cannot be inferred from these findings, further research is warranted to determine the safety and effectiveness of inhaled beta agonists for patients with CHF. PMID- 15523328 TI - Contemporary partial denture designs. 1958. PMID- 15523329 TI - Intrusion and reversal of a free-standing natural tooth bounded by two implant supported prostheses: a clinical report. AB - Although intrusion of natural tooth abutments in tooth-implant connected fixed prostheses has been reported, it can also occur to a free-standing natural tooth bounded by implant prostheses. For the patient described in this article, intrusion was noted with a natural tooth bounded by 2 implant-supported prostheses 5 months after insertion of the prostheses. The intrusion was reversed completely after 5 months with appropriate management. The course of treatment and possible mechanisms of intrusion are provided. PMID- 15523330 TI - Patient-fabricated denture prostheses: a clinical report. AB - This article describes 2 patients who presented with self-made dentures. Each patient had a different motivation for avoiding professional dental treatment. One patient made her own denture because she could not afford dental care, while the other could afford such care but avoided treatment due to a dental phobia. A discussion of diagnosis and treatment planning is presented for each situation. PMID- 15523331 TI - Simultaneous functional and fixed appliance therapy for growth modification and dental alignment prior to prosthetic habilitation in hypohidrotic ectodermal dysplasia: a clinical report. AB - This clinical report describes the simultaneous use of functional and fixed appliances to modify the pattern of dentofacial development and align teeth in preparation for prosthodontic habilitation of a growing child with hypohidrotic ectodermal dysplasia. The treatment objective was to create a more favorable starting point for the prosthodontic phase of habilitation by improving the sagittal and vertical skeletal relationships and facial esthetics. This was accomplished through growth modification with functional appliances conducted simultaneously with eruption of maxillary and mandibular molars, dental arch expansion, alignment, and space management using fixed orthodontic appliances. Orthodontic retention was accomplished by means of removable partial dentures. A second phase of orthodontics conducted closer to the age of skeletal maturation will aim at definitive tooth alignment in preparation for dental implant supported restorations. PMID- 15523332 TI - Finite element analysis of three designs of an implant-supported molar crown. AB - STATEMENT OF PROBLEM: The optimal method of implant support for a single mandibular molar crown is controversial because commonly used, threaded, root form implants developed by Branemark were not originally designed to support individual crowns. PURPOSE: The purpose of this study was to develop a finite element model of a single mandibular first molar crown supported by (1) a standard 3.75-mm-diameter implant, (2) a 5-mm, wide-diameter implant, and (3) double standard-diameter implants, and to compare the induced displacements as a result of various loading conditions. MATERIAL AND METHODS: Three-dimensional finite element models were made to simulate the 3 single-molar implant designs. Each model was analyzed with 2 force magnitudes (35 N and 70 N) and with 2 force directions (vertical and 15 degrees to the vertical axis). Displacements in 3 dimensional space as a result of the simulated loading conditions were evaluated along 3 primary axes, mesiodistal, faciolingual, and superior-inferior. RESULTS: Mesiodistal and buccolingual displacements for the crown supported by the 5-mm diameter implant were reduced by approximately 50% compared with the crown supported by the 3.75-mm implant when the crowns were loaded at the distobuccal cusp tip or the distal marginal ridge. The double-implant design recorded the least mesiodistal displacement with off-center loading of the crown. CONCLUSION: When the crown was loaded off-center, the double-implant design produced substantially less displacement when compared with either of the single-implant designs. PMID- 15523333 TI - Analysis of strain around endosseous dental implants opposing natural teeth or implants. AB - STATEMENT OF PROBLEM: Natural teeth and implants have different force transmission characteristics to bone. PURPOSE: The purpose of this study was to compare strains induced around a natural tooth opposing an implant with strains around occluding implants under static and dynamic loads. MATERIAL AND METHODS: Occlusion was created between a natural molar tooth and an implant in 1 side, and 2 implants in the contralateral side of acrylic resin models of both jaws. Strain gauges were bonded around the neck of the natural tooth and implants, and strains were measured under 75 N and 100 N static axial and lateral dynamic loads in separate load situations using a data acquisition system at sample rate of 1000 Hz. The strain data of the natural tooth and implants were compared for each load by Kruskal-Wallis testing followed by the multiple comparison test (alpha=.05). RESULTS: Compressive strains were induced around natural tooth and implants as a result of static axial loading, whereas combinations of compressive and tensile strains were observed during lateral dynamic loading. Strains around the natural tooth were significantly lower than the opposing implant and occluding implants in the contralateral side for most regions under all loading conditions ( P <.05). There was a general tendency for increased strains around the implant opposing natural tooth under higher loads and particularly under lateral dynamic loading ( P <.05). CONCLUSION: Under static and dynamic loads, strain magnitudes around a natural tooth were significantly lower than that of an opposing implant and occluding implants in the contralateral side. PMID- 15523334 TI - A review of selected dental literature on evidence-based treatment planning for dental implants: report of the Committee on Research in Fixed Prosthodontics of the Academy of Fixed Prosthodontics. AB - This literature review summarizes research with the aim of providing dentists with evidence-based guidelines to apply when planning treatment with osseointegrated implants. Peer-reviewed literature published in the English language between 1969 and 2003 was reviewed using Medline and hand searches. Topics reviewed include systemic host factors such as age, gender, various medical conditions, and patient habits, local host factors involving the quantity and quality of bone and soft tissue, presence of present or past infection and occlusion, prosthetic design factors, including the number and arrangement of implants, size and coatings of implants, cantilevers and connections to natural teeth, and methods to improve outcomes of implant treatment in each category. The review demonstrated that there is no systemic factor or habit that is an absolute contraindication to the placement of osseointegrated implants in the adult patient, although cessation of smoking can improve outcome significantly. The most important local patient factor for successful treatment is the quality and quantity of bone available at the implant site. Specific design criteria are provided, including guidelines for spacing of implants, size, materials, occlusion, and fit. Limitations in the current body of knowledge are identified, and directions for future research are suggested. PMID- 15523335 TI - Effects of varied dimensions of surgical guides on implant angulations. AB - STATEMENT OF PROBLEM: Fabrication of a proper surgical guide is critical for success of implant restorations. The effects of the dimensional factors of the surgical guide on implant placement have not been studied. PURPOSE: The purpose of this study was to determine the effect of varied dimensions (diameter, length, and distance between the underside of the surgical guide and the implant recipient site) of a surgical guide on the accuracy of implant angulation. MATERIAL AND METHODS: In this in vitro study, 240 implant recipient sites were randomly prepared using varied dimensions of a surgical guide. The varied dimensions of the surgical guide's channel and distance were: channel diameter (2, 3, 4, or 5 mm), channel length (6 or 9 mm), and distance between the underside of the surgical guide and the simulated implant recipient site (2 or 4 mm). From these varying dimensions and distances, 16 combinations of dimensions and distances were tested. For each combination, 15 simulated implant recipient site (SIRS) specimens were prepared. The deviated angulation (DA) from the midpoint of the top surface of the 1- x 1-inch simulated implant recipient site (each simulated implant recipient acrylic block contained 5 SIRS of 1 x 1 inch), in the right-to-left (DA(RL)) and front-to-back (DA(FB)) directions, were measured in degrees using a protractor. The data was analyzed using factorial analysis of variance and Tukey's HSD test (alpha=.05). RESULTS: The DA(RL) , in degrees, at a channel length of 9.0 mm (2.33 +/- 1.27) was significantly smaller than at a channel length of 6.0 mm (3.0 +/- 1.42, P =.0001). The DA(RL) , in degrees, at a distance of 4.0 mm (2.13 +/- 1.16) was significantly smaller than at a distance of 2.0 mm (3.16 +/- 1.39, P =.0001). Also, a significant interaction for DA(RL) was found between diameter and distance ( P <.05). For DA(FB) , the varying diameters ( P <.05), lengths ( P =.0001), and distances ( P =.0001) showed significant differences. The DA(FB) at a channel length of 9.0 mm (2.56 degrees +/- 1.51) was significantly smaller than that at 6.0 mm (3.82 degrees +/- 1.87). Significant interactions found for DA(FB) were: diameter by length ( P =.0001), diameter by distance (F=4.547, P =.004), and length by distance (F=11.512, P =.001). CONCLUSION: Within the limitations of this study, the results suggest channel length as the primary controlling factor in minimizing deviated angulations. PMID- 15523336 TI - An evaluation of impression techniques for multiple internal connection implant prostheses. AB - STATEMENT OF PROBLEM: Movement of impression copings inside the impression material using an open-tray impression technique during clinical and laboratory phases may cause inaccuracy in transferring the 3-dimensional spatial orientation of implants intraorally to the definitive cast. Consequently the restoration may require corrective procedures. PURPOSE: This in vitro study evaluated the accuracy of 3 different impression techniques using polyether impression material to obtain a precise definitive cast for a multi-unit implant restoration with multiple internal connection implants. MATERIAL AND METHODS: A reference acrylic resin model with 4 internal connection implants (3i Implant Innovations) was fabricated. Forty-five medium-consistency polyether impressions (Impregum Penta) of this model were made with square impression copings using an open-tray technique. Three groups of 15 specimens each were made with different impression techniques: in the first group, nonmodified square impression copings were used (NM group); in the second group, square impression copings were used and joined together with autopolymerizing acrylic resin before the impression procedure (R [resin] group); and in the third group, square impression copings previously airborne-particle abraded and coated with the manufacturer-recommended impression adhesive were used (M [modified] group). Matching implant replicas were screwed into the square impression copings in the impressions. Impressions were poured with ADA type IV stone (New Fujirock). A single calibrated examiner blinded to the nature of the impression technique used examined all definitive casts to evaluate the positional accuracy (mum) of the implant replica heads using a profile projector (at original magnification x10). These measurements were compared to the measurements calculated on the reference resin model which served as control. Data were analyzed with a 1-way analysis of variance at alpha=.05, followed by the Student Newman-Keuls test (alpha=.05). RESULTS: The data obtained with the profile projector revealed significant differences within the 3 impression techniques ( P <.001). The Student Newman-Keuls procedure disclosed significant differences between the groups, with group R casts being significantly more accurate than group NM and group M casts ( P =.05). The mean distance (+/-SD) between the posterior implants compared to the reference acrylic resin model was 18.17 mum (+/- 6.4) greater for group R casts, 41.27 mum (+/- 8.4) greater for group M casts, and 46.21 mum (+/- 8.9) greater for group NM casts. Distances between the anterior implants were also greater than those recorded on the reference model. The distance was 15.23 mum (+/- 5.9) greater on group R casts, 38.17 mum (+/- 8.3) greater on group M casts, and 43.23 mum (+/- 8.7) greater on group NM casts. CONCLUSION: Within the limitations of this study, improved accuracy of the definitive cast was achieved when the square impression copings joined together with autopolymerizing acrylic resin were used to make an impression of multiple internal connection implants. PMID- 15523337 TI - Spectrophotometric evaluation of the optical influence of different metal alloys and porcelains in the metal-ceramic complex. AB - STATEMENT OF THE PROBLEM: Color matching between natural teeth, shade guides, and metal-ceramic restorations is a common clinical problem. Difficulties related to color matching arise from structural differences that exist between metal-ceramic crowns and natural teeth, the limited range of available ceramic shades, inadequate shade guides, different types of metal alloys, and varying compositions of ceramic materials. PURPOSE: The aim of this study was to investigate the influence of various metal alloys and 2 porcelains on the final color of metal-ceramic complex. Material and methods Four commercial alloys for metal-ceramic restorations, a Ni-Cr (Thermobond), a Co-Cr (Wirobond), a Pd-rich noble (Cerapal-2), and a high noble Au-alloy (V-Delta) were combined with 2 porcelains (Vita Omega and Ceramco Silver) in metal-ceramic specimens with a standardized thickness of layers. Five disc-shaped (10 x 1 mm) specimens were prepared for each alloy/porcelain combination. Only opaque and dentin layers were applied (shade A3). The specimens were analyzed with a spectrophotometer, and data were obtained in the CIE Lab color system. The recorded data were analyzed with a 2-way multiple analysis of variance , a pair-wise comparison of group means (Student's t test), and finally, a categorical regression analysis of variance (CATREG) (alpha=.01). RESULTS: The types of alloy substrate and overlying porcelain significantly affected the color (P < .01). Au and Co-Cr alloys were found to be brighter (higher L* values) than the Ni-Cr and the Pd alloys ( P <.01, SE 0.239). Ceramco porcelain was found to be most red (higher a* values) of all tested alloys ( P <.01). Gold and Pd alloys caused a yellow shift to the metal-ceramic color compared to the Ni-Cr and the Co-Cr alloys with both porcelains ( P < .01, SE 0.165). The detected color differences were visually perceptible for some alloy-porcelain combinations. CONCLUSIONS: The final color of metal ceramic specimens was influenced both from the type of alloy substructure and from the type of overlying porcelain. PMID- 15523338 TI - Comparison of three systems for the polishing of an ultra-low fusing dental porcelain. AB - STATEMENT OF THE PROBLEM: With the introduction of newer dental porcelains, there exists the need to evaluate different porcelain polishing systems available on the market. PURPOSE: The purpose of this study was to compare the surface roughness produced by 3 different porcelain polishing systems on an ultra-low fusing porcelain. MATERIAL AND METHODS: Sixty-three ultra-low fusing porcelain (Finesse) discs (10 x 2 mm) were fabricated and randomly divided into 3 groups (n=21). Both sides of each disc were abraded with a medium-grit diamond bur. One side was autoglazed and was considered a control. The other side was polished until the surface appeared shiny to the naked eye using 1 of 3 porcelain polishing kits (Axis Dental, Jelenko, and Brasseler systems). The surface of each disc was evaluated quantitatively with surface profilometry and qualitatively using scanning electron microscopy (SEM). A mean roughness profile (Ra) value was determined for each side of each specimen to describe the overall roughness of the surface. The Ra mean difference for each specimen was determined by subtracting the mean experimental readings (polished surface) from the mean control readings (glazed surface) and was used for the statistical analysis. The data were analyzed using analysis of variance followed by a Tukey multiple comparison test (alpha=.05). Representative specimens from each group were evaluated by scanning electron microscopy. RESULTS: The Axis porcelain polishing system produced a smoother surface than the Brasseler or Jelenko systems (0.586 +/- 0.256, 0.306 +/- 0.238, and 0.277 +/- 0.230, respectively). No significant difference was found between the Jelenko and Brasseler porcelain polishing kits (F=10.6, P <.001). The images obtained through SEM were evaluated and found to be consistent with the profilometer readings. CONCLUSION: Within the limitations of this study, all 3 porcelain polishing systems produced a surface smoother than the autoglazed surface of Finesse. The Axis system provided a significantly smoother surface compared to the Brasseler and Jelenko polishing systems. PMID- 15523339 TI - In vitro fracture strength of teeth restored with different all-ceramic crown systems. AB - STATEMENT OF PROBLEM: There is insufficient knowledge of the strength of all ceramic crowns bonded to natural teeth to warrant the use of all-ceramic crowns in place of metal-ceramic crowns. PURPOSE: The purpose of this study was to evaluate and compare fracture resistance of crowns made of 3 different types of 2 all-ceramic crown systems-0.4-mm and 0.6-mm aluminum oxide coping crowns and zirconia ceramic coping crowns-and metal-ceramic crowns. MATERIAL AND METHODS: Forty intact, noncarious human maxillary central incisors were divided into 4 groups (n=10): Group MCC (control), metal-ceramic crown (JRVT High Noble Alloy); Group AC4, crown with 0.4-mm aluminum oxide coping (Procera AllCeram); Group AC6, crown with 0.6-mm aluminum oxide coping (Procera AllCeram); and Group ZC6, crown with 0.6-mm zirconia ceramic coping (Procera AllZirkon). Teeth were prepared for complete-coverage all-ceramic crowns so that a final dimension of 5.5 +/- 0.5 mm was achieved incisocervically, mesiodistally, and faciolingually. A 1.0-mm deep shoulder finish line was used with a rounded internal line angle. All restorations were treated with bonding agent (Clearfil SE Bond) and luted with phosphate-monomer-modified adhesive cement (Panavia 21). Fracture strength was tested with a universal testing machine at a crosshead speed of 2 mm per minute with an angle of 30 degrees to the long axis of the tooth after restorations were stored in 100% relative humidity of a normal saline solution for 7 days. The mode of fracture was examined visually. Means were calculated and analyzed with 1-way ANOVA and Tukey's HSD (alpha=.05). RESULTS: The means of fracture strength were: Group MCC, 405 +/- 130 N; Group AC4, 447 +/- 123 N; Group AC6, 476 +/- 174 N; and Group ZC6, 381 +/- 166 N. There was no significant difference between groups ( P =.501). The mode of failure for all specimens was fracture of the natural tooth. CONCLUSIONS: There was no significant difference in the fracture strength of the teeth restored with all-ceramic crowns with 0.4- and 0.6-mm aluminum oxide copings, 0.6-mm zirconia ceramic copings, and metal ceramic crowns. PMID- 15523340 TI - Clinical application of surgical fixation screws in implant prosthodontics--Part II: Indexing implant position. AB - The accurate transfer of intraoral implant position to the definitive cast in situations of immediate implant loading in completely edentulous patients is an important step and may be difficult to achieve with common transfer and impression methods. Intraoral fixed points of reference accurately transferred to the definitive cast are necessary to provide precise and stable placement of radiographic and surgical templates during the presurgical diagnostic phase, during implant placement, and during fabrication of an immediate provisional restoration. This article describes a technique using surgical microfixation screws to provide fixed reference points and transfer the position of the conventional implants to the definitive cast. PMID- 15523341 TI - Repair of bis-acryl provisional restorations using flowable composite resin. AB - Provisional restorations provide interim coverage for prepared teeth while fixed definitive restorations are fabricated. Several types of autopolymerizing acrylic resins have been used for many years to fabricate provisional restorations. In recent years, bis-acryl resin composite material has gained popularity among clinicians for the direct fabrication of provisional fixed restorations. Occasionally, deficiencies may occur while fabricating a direct provisional restoration and require chairside repair. This article describes an effective procedure for the use of light-polymerized flowable composite resin for the intraoral repair of bis-acryl provisional restorations. PMID- 15523342 TI - Stable interocclusal records for implant patients with posterior edentulism. PMID- 15523343 TI - Alternative technique for investing abutments for screw-retained implant supported restorations. PMID- 15523344 TI - Amalgam repair with a 4-META resin. PMID- 15523345 TI - DNA microarrays: from structural genomics to functional genomics. The applications of gene chips in dermatology and dermatopathology. AB - The human genome project was successful in sequencing the entire human genome and ended earlier than expected. The vast genetic information now available will have far-reaching consequences for medicine in the twenty-first century. The knowledge gained from the mapping and sequencing of human genes on a genome-wide scale- commonly referred to as structural genomics--is prerequisite for studies that focus on the functional aspects of genes. A recently invented technique, known as gene chip, or DNA microarray, technology, allows the study of the function of thousands of genes at once, thereby opening the door to the new field of functional genomics. At its core, the DNA microarray utilizes a unique feature of DNA known as complementary hybridization. As such, it is not different from Southern (DNA) blot or northern (RNA) blot hybridizations, or the polymerase chain reaction, with the exception that it allows expression profiling of the entire human genome in a single hybridization experiment. The article highlights the principles, technology, and applications of DNA microarrays as they pertain to the field of dermatology and dermatopathology. The most important applications are the gene expression profiling of skin cancer, especially of melanoma. Other potential applications include gene expression profiling of inflammatory skin diseases, the mutational analysis of genodermatoses, and polymorphism screening, as well as drug development and chemosensitivity prediction. cDNA microarrays will shape the diagnostic approach of the dermatology and the dermatopathology of the future and may lead to new therapeutic options. PMID- 15523346 TI - Measurement of the severity of rosacea. AB - BACKGROUND: There are no standardized measures for the severity of rosacea. OBJECTIVES: To determine what clinical signs correlate best with global severity of rosacea, and to examine inter-rater reliability. Methods Four clinicians each made 82 assessments of rosacea patients. Each assessment used 60 0-to-10 Likert like scales for the signs of rosacea. Subjects also assessed the severity of their rosacea. RESULTS: The clinicians' assessment of global severity correlated strongly with erythema, especially on the cheeks. Subjects' assessment of global severity correlated more strongly with papules/pustules. Different methods of assessing severity--estimation of area involved, intensity, or lesion counts--did not produce significantly different results. Inter-rater reliability was low on 11-point (0-10) scales, but improved when scales were collapsed to 5 or 4 points. CONCLUSIONS: Clinicians and patients assess severity of rosacea differently, with clinicians focusing on erythema and patients focusing on papules/pustules. New instruments for assessing severity must address inter-rater reliability. PMID- 15523347 TI - Determinants of quality of life in patients with psoriasis: a study from the US population. AB - BACKGROUND: Psoriasis is a common disease with substantial effects on quality of life. Few quality of life studies have been performed in psoriasis patients from the general US population. OBJECTIVE: To describe the determinants of quality of life in psoriasis patients from the US population. METHODS: Patients were randomly selected from the US population. Patients who identified themselves as having been diagnosed with psoriasis by a physician were invited to complete a more detailed survey about quality of life. RESULTS: Two hundred sixty-six psoriasis patients from the US population completed the detailed survey. Body surface area showed the strongest association with decrements in quality of life (Spearman 0.50, P < .0001). Younger patients and female patients also had statistically significant reductions in quality of life. Increasing psoriasis severity was associated with seeking care from multiple physicians and having decrements in income. CONCLUSION: Patients with more extensive skin involvement have greater reductions in quality of life. Female patients and young patients are affected to a greater extent. PMID- 15523349 TI - Characteristics of mycobacterial infection in patients with immunodeficiency and nuclear factor-kappaB essential modulator mutation, with or without ectodermal dysplasia. AB - Hypomorphic mutations of the nuclear factor kappaB essential modulator gene cause ectodermal dysplasia and immunodeficiency. Affected patients have increased susceptibility to mycobacterial disease including cutaneous manifestations. We describe clinical and histopathologic characteristics of 5 patients with nuclear factor kappaB essential modulator gene mutations and mycobacterial infections, two of whom had mycobacterial cutaneous infections. PMID- 15523348 TI - Effect of a single course of isotretinoin therapy on bone mineral density in adolescent patients with severe, recalcitrant, nodular acne. AB - BACKGROUND: Adverse changes in bone have been reported for patients undergoing high-dose, long-term (several years) isotretinoin therapy for disorders of cornification. The effect of short-term (4-5 months) therapy at the lower dose recommended for acne on bone development in younger, growing adolescent (12-17 years) patients has not been well studied. OBJECTIVE: The purpose of the study was to evaluate the effect of a standard, single course of isotretinoin (Accutane) therapy on bone mineral density (BMD) of the lumbar spine and hip in adolescents ages 12 to 17 years with severe, recalcitrant, nodular acne. METHODS: In this open-label, multicenter study, 217 adolescents (81 girls) with severe, recalcitrant, nodular acne were enrolled and treated with isotretinoin twice daily with food at the recommended total dose of approximately 1 mg/kg for 16 to 20 weeks. BMD in the lumbar spine and hip was measured at baseline and at the end of therapy by dual energy radiograph absorptiometry. RESULTS: There was no clinically significant mean change in BMD measured at the lumbar spine (+1.4%, range: -4.9% to +12.3%) or total hip (-0.26%, range: -11.3% to +15.0%). Hyperostosis was not observed in any patient. Typical efficacy expected in the treatment of acne was observed. CONCLUSIONS: A 16- to 20-week course of isotretinoin treatment at the recommended dose for severe acne has no clinically significant effect on lumbar spine and total hip BMD in the adolescent (12-17 years) population. PMID- 15523350 TI - Tacrolimus ointment is effective for facial and intertriginous psoriasis. AB - BACKGROUND: Intertriginous and facial involvement are manifestations of psoriasis that require a different approach than is used for typical plaque psoriasis on other skin areas. Topical corticosteroids are the primary treatment for psoriasis; however, the side effects of corticosteroids are magnified on intertriginous and facial skin. Topical tacrolimus offers the potential for anti inflammatory effect without the atrophy or other local side effects associated with the use of topical corticosteroids. OBJECTIVE: To determine the efficacy and tolerability of 0.1% tacrolimus ointment for the treatment of facial or intertriginous psoriasis. METHODS: One hundred sixty-seven patients 16 years or older were evaluated in an 8-week, randomized, double-blind, vehicle-controlled, multi-center study. Upon entry into the study, patients were randomized 2:1 to apply the tacrolimus ointment 0.1% or vehicle twice daily to all psoriatic lesions of the face or intertriginous areas for 8 weeks. The physician's global assessment was used to assess improvement from baseline. The inverse psoriasis severity for patients was measured using a 6-point scale from clear to very severe. RESULTS: As early as day 8, more patients ( P = .004) had cleared or achieved excellent improvement in the 0.1% tacrolimus ointment group compared to the vehicle group (24.8% vs 5.8%). At the end of the 8-week treatment period 65.2% of the tacrolimus ointment group and 31.5% of the vehicle were clear or almost clear ( P < .0001) based on a Static Severity Score. Adverse events were similar in the 0.1% tacrolimus ointment and vehicle groups. Conclusion Tacrolimus ointment is an effective treatment for psoriasis of the face or intertriginous areas. PMID- 15523351 TI - Pimecrolimus cream 1% in the treatment of intertriginous psoriasis: a double blind, randomized study. AB - BACKGROUND: Inverse psoriasis can be difficult to treat because of the high sensitivity of intertriginous areas to cutaneous side effects, such as irritation and striae. Pimecrolimus, a well-tolerated, nonatrophogenic, skin-selective inflammatory cytokine inhibitor, has been shown to be effective in the treatment of psoriasis when applied topically under occlusion. OBJECTIVE: This study evaluated the efficacy and safety of pimecrolimus cream 1% versus vehicle twice a day in the treatment of inverse psoriasis. Methods This was a double-blind, randomized, vehicle-controlled study in 57 patients with moderate to severe inverse psoriasis. Patients were evaluated using Investigator's Global Assessment of overall severity, Target Area Score, and Patient Self-Assessment. RESULTS: A significant between-group difference was observed early on, with 54% of the pimecrolimus group versus 21% of the vehicle group having an Investigator's Global Assessment score of 0 or 1 (clear or almost clear) at week 2 ( P = .0169). By week 8, 71% of the pimecrolimus group had an Investigator's Global Assessment score of 0 or 1. Change from baseline in Target Area Score was -2.4 (pimecrolimus group) compared with -0.7 (vehicle) at day 3 ( P < .0001). By week 8, 82% of patients using pimecrolimus scored their disease as well or completely controlled versus 41% of the vehicle group ( P = .0007). Adverse events were similar between groups. CONCLUSION: Pimecrolimus cream 1% is an effective treatment for inverse psoriasis with a rapid onset of action, and is safe and well-tolerated. PMID- 15523352 TI - Sweat gland morphology and periglandular innervation in essential palmar hyperhidrosis before and after treatment with intradermal botulinum toxin. AB - BACKGROUND: Intradermal botulinum toxin (Btx) produces long-lasting relief of focal hyperhidrosis, but its mechanism of action is poorly understood. OBJECTIVE: To study the effect of Btx A on the size and innervation of sweat glands in patients with palmar hyperhidrosis. METHODS: Palmar skin biopsy was performed in 26 hyperhidrotic patients before scheduled Btx treatment and in 11 controls. Twelve of the patients also underwent biopsy 1 to 6 months after the Btx injections. Sweat gland morphology was investigated by light microscopy; the cross-sectional area of the secretory tubule and its lumen was measured by image analysis. Immunofluorescence (IF) with antibodies to the neural markers protein gene product 9.5 (PGP 9.5) and growth-associated protein 43 (GAP 43), and to vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP), was used to analyze the periglandular innervation. RESULTS: The gross morphology of the sweat glands was similar in patients and controls, with no significant differences in tubular and luminal areas between the groups. After Btx treatment, the tubular dimensions remained unchanged, but the lumen tended to be smaller ( P = .07). Around the glands, increased GAP 43 staining indicating sprouting was seen within 3 months after Btx treatment ( P = .016); whereas the PGP 9.5 staining was decreased in most specimens ( P = .09) indicating lack of functional nerve growth. No change in VIP or CGRP immunoreactivity was observed. CONCLUSIONS: The sweat glands appear structurally normal in hyperhidrotic patients before Btx therapy, whereas after therapy the luminal area of the gland is frequently diminished. The IF data GAP 43/PGP 9.5 suggest that Btx therapy induces long-standing functional denervation of the sweat glands, which might explain its anti-transpiratory efficacy. PMID- 15523353 TI - Reduction of treatment frequency and UVA dose does not substantially compromise the antipsoriatic effect of oral psoralen-UVA. AB - BACKGROUND: The carcinogenic potential of 8-methoxypsoralen photochemotherapy (psoralen-UVA [PUVA]) is correlated with the total number of treatments and cumulative UVA dose applied during oral PUVA therapy. OBJECTIVE: We sought to determine whether reducing treatment frequency and UVA dose affects the therapeutic efficacy of oral PUVA for patients with chronic plaque psoriasis. METHODS: This was a prospective, randomized, half-side study performed in a photodermatology department in a dermatology hospital. Eighteen consecutive patients with chronic plaque psoriasis received paired PUVA regimens (0.5 minimal phototoxic dose [MPD] 4 times/wk vs 1 MPD twice/wk, 0.5 MPD twice/wk vs 1 MPD twice/wk, and 0.5 MPD twice/wk vs 0.75 MPD twice/wk). The PUVA regimens were assessed for reduction of Psoriasis Area and Severity Index (PASI) score and the number of treatments and cumulative UVA dose required to reduce PASI score to defined end points (ie, PASI reductions of 25%, 50%, and 75%) or to induce complete remission (PASI < 3). RESULTS: Reducing the number of treatments while maintaining the same UVA dose per week did not reduce overall therapeutic efficacy. Reducing the number of treatments to twice a week and reducing the UVA dose from 1 MPD to 0.75 or 0.5 MPD per treatment only slightly affected intermediate therapeutic efficacy (between the second and seventh weeks of therapy) but had no effect on final clearance rates. The time to complete clearance did not significantly differ between regimens. The mean cumulative UVA dose was significantly lower for the least intensive dose regimen (0.5 MPD twice/wk) than for the more intensive regimens. CONCLUSIONS: Reducing treatment frequency and UVA dose does not substantially compromise the therapeutic efficacy of PUVA. PMID- 15523354 TI - Digital anesthesia with epinephrine: an old myth revisited. AB - BACKGROUND: The prohibition against the use epinephrine with local anesthetics for digital blocks or infiltrative anesthesia is an established dogma in dermatologic surgery. Major textbooks reinforce this teaching suggesting that there is substantial risk of digital gangrene caused by local anesthesia containing epinephrine. OBJECTIVE: To provide a comprehensive literature review of the cases of digital necrosis associated with the use of local anesthesia containing epinephrine. METHODS: A PubMed search of the National Library of Medicine database using the terms "lidocaine" and "epinephrine" and "finger" with no specified limits was performed. RESULTS: A total of 16 papers were referenced and only 6 papers dealt with digital anesthesia. A total of 50 cases of digital gangrene were reported, mostly in the early part of the 20th century. In 21 cases digital gangrene was associated with anesthetic mixed with epinephrine. Actual concentration of epinephrine was known in only 4 cases. Careful analysis of all cases of necrosis did not support epinephrine itself as a cause. Other contributing factors including older compounds (cocaine, eukaine, and procaine), non-standardized inaccurate methods of mixing epinephrine with lidocaine, inappropriate use of a tourniquet, postoperative hot soaks, infection, or large anesthetic volume were also present. None of the reported cases were associated with the use of a commercial lidocaine-epinephrine mixture. CONCLUSION: A literature review failed to provide evidence to support the dogma that block or infiltrative anesthesia with lidocaine and epinephrine produces digital necrosis. Proper injection technique and adequate selection of patients (absence of thrombotic, vasospastic conditions, or uncontrolled hypertension) are mandatory to minimize complications. The addition of epinephrine, in fact, reduces the need for the use of tourniquets and large volumes of anesthetic and provides better and longer pain control during digital procedures. PMID- 15523355 TI - Tacrolimus ointment promotes repigmentation of vitiligo in children: a review of 57 cases. AB - BACKGROUND: Vitiligo is an autoimmune disorder characterized by loss of pigmentation. Phototherapy and application of topical corticosteroids are most commonly prescribed. However, these therapies are often not effective and use of corticosteroids on the face may lead to cutaneous atrophy, telangiectasia, and ocular complications. OBJECTIVE: We sought to assess the efficacy of topical tacrolimus ointment in the treatment of pediatric vitiligo. METHODS: A retrospective review was performed of 57 pediatric patients with vitiligo at two clinical sites. Patients were treated with tacrolimus ointment for at least 3 months. Clinical responses were documented during clinic visits, and by pretacrolimus and posttacrolimus photography. RESULTS: At least partial response was noted to tacrolimus ointment on the head and neck in 89%, and on the trunk and extremities in 63% of patients. Facial vitiligo of the segmental type showed the best response rate. Two patients initially experienced burning on application. CONCLUSIONS: Topical tacrolimus ointment is an effective alternative therapy for childhood vitiligo, particularly involving the head and neck. PMID- 15523356 TI - Extramammary Paget's disease: surgical treatment with Mohs micrographic surgery. AB - BACKGROUND: Extramammary Paget's disease (EMPD) is an uncommon tumor that has a high rate of recurrence after conventional surgical treatments. OBJECTIVE: Our purpose was to establish the efficacy of Mohs micrographic surgery (MMS) in the treatment of EMPD, and to make treatment recommendations with regard to surgical margins. We also attempted to summarize the published recurrence rates of EMPD after standard surgical management. METHODS: In a retrospective chart review, pertinent demographic data, tumor data, treatment characteristics, and follow-up data were tabulated. A search of the literature for recurrence rates after MMS and non-MMS surgical treatment modalities was performed. RESULTS: The recurrence rate after treatment with MMS was 16% for primary EMPD and 50% for recurrent EMPD. The 5-year tumor-free rates (Kaplan-Meier analysis) were 80% for primary tumors and 56% for recurrent tumors. Using MMS, the salvage rate (and, hence, overall cure rate) was 100%. Margins of 5 cm were required to clear 97% of the tumors. The recurrence rate after non-MMS (from the literature) is 33% to 60%. CONCLUSION: MMS is effective, and superior to standard surgical management in the treatment of EMPD. We recommend a 5-cm margin of normal skin if MMS cannot be offered. PMID- 15523357 TI - Reticulate erythema following diode laser-assisted hair removal: a new side effect of a common procedure. AB - The popularity of laser-assisted hair removal has grown rapidly since April 3, 1995 when the Food and Drug Administration approved the introduction of the first hair removal laser system. Lasers with wavelengths in the red and infrared portion of the electromagnetic spectrum are most often used for hair removal because they effectively target melanin in the hair follicle and can potentially penetrate to the appropriate depth of the dermis. Despite all efforts to protect the skin from damage, photoepilation may result in clinically significant adverse reactions. The most common and known side effects of laser hair removal include transient erythema, perifollicular edema, pain, folliculitis, hyper-pigmentation, hypopigmentation, crusting, purpura, erosions and scarring. The present report describes the appearance of a reticulate erythema after diode laser treatment for hair removal, encountered in 10 patients in our clinics in London and Israel. To the best of our knowledge, this is the first report of this side effect. The aim of this work is to detail the clinical manifestations, histological findings, and follow-up of these patients in order to expand the clinical spectrum of laser assisted hair removal side effects and to alert dermatologists to the possibility of this type of net-like erythema. PMID- 15523358 TI - Localized cutaneous posttransplant Epstein-Barr virus-associated lymphoproliferative disorder. AB - Localized cutaneous posttransplant lymphoproliferative disorder is a rare complication of solid organ transplantation. Biopsies demonstrate diffuse dermal proliferations of atypical lymphocytes with variable Epstein-Barr virus latent membrane protein-1 expression. Extracutaneous involvement is absent. PMID- 15523359 TI - Scientific and regulatory issues related to indoor tanning. PMID- 15523360 TI - Skin diseases associated with Malassezia species. AB - The yeasts of the genus Malassezia have been associated with a number of diseases affecting the human skin, such as pityriasis versicolor, Malassezia (Pityrosporum) folliculitis, seborrheic dermatitis and dandruff, atopic dermatitis, psoriasis, and--less commonly--with other dermatologic disorders such as confluent and reticulated papillomatosis, onychomycosis, and transient acantholytic dermatosis. Although Malassezia yeasts are a part of the normal microflora, under certain conditions they can cause superficial skin infection. The study of the clinical role of Malassezia species has been surrounded by controversy because of their fastidious nature in vitro, and relative difficulty in isolation, cultivation, and identification. Many studies have been published in the past few years after the taxonomic revision carried out in 1996 in which 7 species were recognized. Two new species have been recently described, one of which has been isolated from patients with atopic dermatitis. This review focuses on the clinical, mycologic, and immunologic aspects of the various skin diseases associated with Malassezia. It also highlights the importance of individual Malassezia species in the different dermatologic disorders related to these yeasts. PMID- 15523361 TI - Petrified ears: calcification of the auricular cartilage. PMID- 15523362 TI - The genetics of melanoma. PMID- 15523364 TI - Surgical pearl: readily available cryosurgery shield. PMID- 15523363 TI - Genetic testing in hereditary melanoma. AB - Advances in our understanding of molecular genetics bring about unique challenges in our ability to practice molecular medicine. With the availability of commercial testing for various genetic cancer syndromes, including hereditary melanoma, specific issues regarding its use and limitations require attention before full translation of this tool into the clinical setting. PMID- 15523365 TI - Dermoscopic assessment of long-term topical therapies with potent steroids in chronic psoriasis. AB - We investigated the value of the dermoscope for monitoring the long term safety of high potency topical steroids in patients with chronic psoriasis. We observed for the first time that the overuse of topical steroids resulted in the appearance of clinically unapparent but dermoscopically apparent "red lines" (linear telangiectasias) in the treated plaques and/or skin adjacent to the treated plaques (P < .03). We concluded that dermoscopy may help reveal the early signs of impending steroid-induced atrophy ("red lines") before they become clinically evident with the naked eye and before the atrophy becomes permanent. PMID- 15523366 TI - Indoor UV tanning operator opinion regarding youth access: an electronic survey. AB - BACKGROUND: Adolescents frequently use indoor tanning facilities, but little is known about tanning facility operator opinions regarding this use. Objective To assess indoor tanning operator attitudes and stated practices regarding youth access. METHODS: We electronically surveyed 89 indoor tanning facilities and 130 spas with active e-mail addresses. RESULTS: The survey response rate was 21% for both indoor tanning facilities (19/89) and spas (27/130). Most tanning operators (92%) felt that a client can be too young to receive indoor tanning, and most felt that written parental permission should be required (80%). Most facilities required adult accompaniment (92%), and most operators had discouraged a customer from receiving indoor tanning based on age (77%). The reported age of the youngest patron receiving indoor tanning ranged from 5 to 21 years. CONCLUSION: Our sample of indoor tanning facility operators believed that minimum age and parental consent regulations for indoor tanning should be required. PMID- 15523367 TI - Therapy of refractory pemphigus vulgaris with monoclonal anti-CD20 antibody (rituximab). AB - Three patients with refractory pemphigus vulgaris improved after rituximab, an anti-CD20 monoclonal antibody directed against B cells. Treatment was well tolerated immediately, but one patient developed fatal pneumocystis carinii pneumonia. Serious infections have occurred in several pemphigus patients treated with rituximab and immunosuppressive medications, warranting further evaluation of risk-benefit ratio. PMID- 15523368 TI - Punctate exanthem of West Nile Virus infection: report of 3 cases. AB - The West Nile Virus (WNV) has rapidly emerged as an important etiology of meningoencephalitis in North America since 1999. Diagnosis of this infection on clinical grounds is difficult, as many signs and symptoms of infection are nonspecific. Although cutaneous manifestations are common in WNV-infected patients, these have not been described in detail nor are clinical images widely available. We describe 3 patients with WNV infections, two ambulatory, one hospitalized, who developed punctate erythematous, macular, and papular eruptions, most pronounced on the extremities. Histopathologic findings in one case showed a sparse superficial perivascular lymphocytic infiltrate, a feature commonly seen in viral exanthems but not previously reported with WNV infection. A literature review provides support that this punctate exanthem is a common cutaneous presentation of WNV infection. PMID- 15523369 TI - Genital ulcers caused by Epstein-Barr virus. AB - In North America, the most common cause of vulvar ulcers is infection with herpes simplex virus. However, Epstein-Barr virus can also cause vulvar ulcers, and may be underrecognized. Unlike herpes simplex virus, Epstein-Barr virus is not necessarily sexually transmitted. Therefore, it is particularly important to include in the differential diagnosis of genital ulcers for patients for whom a diagnosis of a venereal infection has important psychosocial consequences. We report three cases of acute primary Epstein-Barr virus infection in which the presenting symptoms were vulvar ulceration. PMID- 15523370 TI - Severe sialadenitis: a new complication of drug reaction with eosinophilia and systemic symptoms. AB - BACKGROUND: Drug cutaneous reaction and isolated cases of parotitis induced by terbinafine have been reported. OBSERVATION: We report a case of drug reaction with eosinophilia and systemic symptoms induced by terbinafine associated with a severe sialadenitis and a complete sicca syndrome. Evolution was protracted with a slow recovery of the rash but sicca syndrome persisted with only a very mild improvement at 6 months. CONCLUSION: Liver, kidneys, lungs, and heart are the organs the most frequently involved in drug reaction with eosinophilia and systemic symptoms. Salivary and lacrimal glands can also be severely involved in the course of drug reaction with eosinophilia and systemic symptoms. PMID- 15523371 TI - Prolonged evolution of a lentigo maligna. AB - Lentigo maligna (LM) is a melanocytic lesion which is a potential precursor to melanoma and often has a prolonged intraepidermal growth phase before evolving into lentigo maligna melanoma (LMM). LM is also noted for its tendency to locally recur after treatment. We present a patient who had a persistent LM on her left cheek which, despite multiple excisions, persisted and transformed into a partially amelanotic LMM roughly three decades later. Our patient's course was also notable for this melanoma recurring at the edge of, and subsequently migrating into, a previously placed skin graft. PMID- 15523372 TI - Klebsiella pneumoniae cellulitis in an immunocompetent man. PMID- 15523373 TI - 308-nm excimer laser therapy in alopecia areata. PMID- 15523374 TI - Amyopathic dermatomyositis presenting during pregnancy. PMID- 15523375 TI - Luxation of the eyeball. PMID- 15523376 TI - Localization of cutaneous sarcoidosis: From trauma to scars. PMID- 15523378 TI - Circumscribed palmar or plantar hypokeratosis: report of two additional cases. PMID- 15523377 TI - Uremic pruritus: still an important clinical problem. PMID- 15523385 TI - Edward D. Churchill and the "rectangular" surgical residency. PMID- 15523386 TI - Residency training in surgery in the 21st century: a new paradigm. PMID- 15523387 TI - Graduate surgical education redesign: reflections on curriculum theory and practice. PMID- 15523389 TI - Surgeons and stem cells: a pragmatic perspective on shifting paradigms. PMID- 15523390 TI - Mortality in sporadic primary hyperparathyroidism: nationwide cohort study of multiple parathyroid gland disease. AB - BACKGROUND: The risk of dying from primary hyperparathyroidism (pHPT) is controversial and has been explored mainly in single parathyroid gland disease. The present study investigates mortality in pHPT due to multiple parathyroid gland disease. METHODS: We used the nationwide Swedish In-patient Register and Cause-of-Death Registry to compare the mortality in 3485 Swedish patients subjected to parathyroidectomy during 1964 to 1999 with that of the Swedish population (standardized for age, gender, and calendar year). The patient cohort includes 36,596 person years. RESULTS: Increased risk of death beyond the first postoperative year (standardized mortality ratio, 1.4; 95% CI, 1.37-1.52) was found in both sexes and for all age intervals below 80 years. The increased risk persisted more than 15 years postoperatively and related to cardiovascular diseases, diabetes mellitus, urogenital diseases, and malignant disorders. The increased risk of dying in cardiovascular diseases normalized during 1990 to 1999. CONCLUSIONS: pHPT caused by multiple parathyroid gland enlargement is associated with an excessive mortality similar to pHPT of single parathyroid adenoma. The findings substantiate that modern modes of surgical treatment for pHPT normalize the risk of dying from cardiovascular complications and that the hyperpararthyroid state per se is the possible cause of the premature death. PMID- 15523391 TI - Excessive alcohol consumption and perioperative outcome. AB - BACKGROUND: Excessive alcohol consumption is a well-recognized factor contributing to premature morbidity and mortality. METHODS: This retrospective, matched cohort study was designed to assess the attributable effects of excessive alcohol consumption on outcome in patients undergoing noncardiac surgery. All data of 28,065 patients operated at a tertiary care university hospital were recorded with a computerized anesthesia record-keeping system. Cases were defined as patients with history of excessive alcohol consumption (>30 g alcohol per day). Controls were selected according to matching variables in a stepwise fashion. RESULTS: In our data set, 928 patients (3.3%) were found with a history of excessive alcohol consumption. Matching was successful in 897 patients (97%). The crude mortality rates for the cases were 1.3% and 1.6%, for the matched controls (P=.084, power=0.85). Prolonged length of hospital stay was observed in 38% versus 33% (P=.013, power=0.50), admission to the intensive care unit was deemed necessary in 11% versus 9% (P=.027, power=0.55), and intraoperative cardiovascular events were detected from the database in 22% versus 21% (P=.053, power=0.61). CONCLUSIONS: In this study, history of excessive alcohol consumption alone is not a factor leading to an increased perioperative risk in noncardiac surgery. PMID- 15523392 TI - Surgical management of neoplasms of the ampulla of Vater: local resection or pancreatoduodenectomy and prognostic factors for survival. AB - BACKGROUND: Neoplasms of the ampulla of Vater have a better 5-year survival than pancreatic and bile duct neoplasms after resection. This study was performed to analyze the outcome after local resection and pancreatoduodenectomy (PD) and to identify predictive factors for survival. METHODS: We used a prospective database to evaluate 145 patients (1992-2002) with a neoplasm of the ampulla of Vater. RESULTS: The median age of the entire cohort was 66 years. Agreement between preoperative biopsies and definite resection specimen was 58% for invasive adenocarcinoma. Local resection was performed in 25 patients, but the operation was adequate therapy in only 16 patients (64%). Subsequent PD (n=9) was performed in the remaining patients because of an R1 resection. Other patients (n=120) underwent an elective PD. Hospital mortality was 4.0% (1/25) after local resection and 5.0% (6/120) after PD. Multivariate analysis revealed that advanced invasion and nodal status were independent predictive factors for survival. The overall 5-year actuarial survival of patients with adenocarcinoma after PD was 37%. CONCLUSIONS: Preoperative biopsies have a poor diagnostic accuracy. Local resection is an adequate surgical treatment for adenomas. In experienced hands, PD is the preferred treatment for patients with adenocarcinoma. PMID- 15523393 TI - A randomized multicenter trial comparing resection and radiochemotherapy for resectable locally invasive pancreatic cancer. AB - BACKGROUND: Though the outcome of resection for locally invasive pancreatic cancer is still poor, it has gradually improved in Japan, and the 5-year survival is now about 10%. However, the advantage of resection over radiochemotherapy has not yet been confirmed by a randomized trial. We conducted this study to compare surgical resection alone versus radiochemotherapy without resection for locally invasive pancreatic cancer using a multicenter randomized design. METHODS: Patients with pancreatic cancer who met our preoperative criteria for inclusion (pancreatic cancer invading the pancreatic capsule without involvement of the superior mesenteric artery or the common hepatic artery, or without distant metastasis) underwent laparotomy. Patients with operative findings consistent with our criteria were randomized into a radical resection group and a radiochemotherapy group (200 mg/m(2)/day of intravenous 5-fluorouracil and 5040 cGy of radiotherapy) without resection. The 2 groups were compared for mean survival, hazard ratio, 1-year survival, quality of life scores, and hematologic and blood chemical data. RESULTS: Twenty patients were assigned to the resection group and 22 to the radiochemotherapy group. There was 1 operative death. The surgical resection group had better results than the radiochemotherapy group as measured by 1-year survival (62% vs 32 %, P=.05), mean survival time (>17 vs 11 months, P < .03), and hazard ratio (0.46, P=.04). There were no differences in the quality of life score or laboratory data apart from increased diarrhea after surgical resection. CONCLUSIONS: Locally invasive pancreatic cancer without distant metastases and major arterial invasion appears to be best treated by surgical resection. PMID- 15523394 TI - Should the extrahepatic bile duct be resected for locally advanced gallbladder cancer? AB - BACKGROUND: The incidence and mode of spread of carcinoma of the gallbladder into the hepatoduodenal ligament have not been well described pathologically for gallbladder carcinoma extending into the subserosa and beyond. METHODS: Between 1985 and 2002, 50 consecutive patients with gallbladder carcinoma extending into the subserosa or beyond underwent radical surgery, including extrahepatic bile duct resection. Serial sections of specimens of the resected extrahepatic bile ducts were examined to determine the incidence and the pattern of invasion of the hepatoduodenal ligament from the primary cancer. RESULTS: Invasion of the hepatoduodenal ligament was present in 30 of the 50 specimens. Of these, 9 showed direct extramural spread (type I), 4 showed continuous intramural spread (type II), 5 showed distant spread separated from the primary tumor (type III), and 4 showed spread of cancer cells from metastatic lymph nodes (type IV). The remaining 8 patients had more than 1 type: 1 patient had types I + III; 3 had types I + III + IV; and 4 had types III+IV. Invasion of the hepatoduodenal ligament was present in 24 of 44 patients without preoperative obstructive jaundice and in 2 of 13 patients with stage IB disease. Patients with types II, III, and IV spread into the hepatoduodenal ligament had significantly better survival than those with type I spread. CONCLUSIONS: Gallbladder carcinoma extending into the subserosa or beyond invades the hepatoduodenal ligament with relatively high frequency. Preoperative diagnosis of this invasion is difficult; therefore, strong consideration should be given to resection of the extrahepatic bile ducts and lymph nodes. PMID- 15523396 TI - Postoperative colonic motility increases after early food intake in patients undergoing colorectal surgery. AB - BACKGROUND: Stimulation of colonic motility by the gastrocolonic response may help to reduce inhibition of gastrointestinal motility after colorectal surgery. We aimed to investigate whether postoperative colonic motility is increased after early food intake. METHODS: Nineteen patients undergoing colorectal surgery and 7 healthy volunteers were investigated. Colonic motility was recorded with a combined manometry/barostat system, and the effect of a standard 500-kcal meal was evaluated once in healthy volunteers and in 15 patients on the first and second postoperative day. Four patients remained unfed, serving as controls. RESULTS: In patients, the colonic motility index increased from 12 +/- 5 at baseline to 65 +/- 24 mm Hg after the meal on postoperative day 1 (mean +/- SEM; P < .01), while barostat bag volumes decreased, indicating a rise in colonic tone. On day 2, the motility index was 62 +/- 17 mm Hg at baseline and did not change after the meal. In unfed controls, no change was observed during colonic motility recordings on both postoperative days. In healthy volunteers, the colonic motility index increased from 98 +/- 52 at baseline to 151 +/- 58 mm Hg postprandially (P < .05). CONCLUSIONS: As in healthy volunteers, there is a potential to stimulate colonic motility by early food intake in postoperative patients. This may help to improve prolonged colonic motility disorders after colorectal surgery. PMID- 15523397 TI - Effect of portal hemodynamics on liver regeneration studied in a novel portohepatic shunt rat model. AB - BACKGROUND: The role of portal hemodynamics on liver regeneration after partial hepatectomy is not fully understood. The aim of our study was to characterize the effects of portal hemodynamics using a novel rat model. METHODS: We established a rat model of a portohepatic shunt with a 70% hepatectomy (PHS model), in which the portal pressure remained stable during and after the 70% hepatectomy. To assess the effect of portal hemodynamics on liver injury and regeneration in the first 24 hours, we compared PHS rats with those with a simple 70% hepatectomy. RESULTS: Biochemical and histopathologic changes were similar between the 2 groups. Liver weight increased in the control, whereas it did not in the PHS group (P = .0021). Hepatocytes were enlarged in the control but not in the PHS group, although DNA synthesis was similar in both groups. Apoptotic hepatocytes increased markedly in PHS at 24 hours, whereas minimal apoptosis was noted throughout the course of the study in the control group. Hepatocyte growth factor increased similarly, except that it was not activated in PHS. CONCLUSIONS: Our results suggested that a portal hyperdynamic state early after a 70% hepatectomy was necessary for liver regeneration through activation of hepatocyte growth factor, promoting hepatocyte hypertrophy and avoiding apoptosis, while DNA synthesis in hepatocytes was independent of portal hemodynamics. PMID- 15523398 TI - Neuroendocrine responses mediate macrophage function after trauma. AB - BACKGROUND: Clearly understanding the interactions between macrophage (M phi) generated inflammatory mediators and the neuroendocrine system in regulating immune function after traumatic injury may aid in reversing trauma-mediated immune dysfunction and diminish the incidence and severity of infection in the traumatized patient. METHODS: Trauma consisted of an open femur fracture and 40% retro-orbital hemorrhage (Trauma) or anesthesia alone (Control). Female Balb/C mice (6-8 weeks) with intact adrenal glands (Intact) or a bilateral adrenalectomy (ADX) were used. For glucocorticoid studies, corticosterone or a vehicle was administered via intraperitoneal (ip) injection 2 hours before the trauma. Splenic M phis were harvested and prostaglandin E(2) (PGE(2)) and interleukin-6 (IL-6) production, and mRNA, cyclooxygenase-2 (COX-2) protein, and nuclear factor kappa B (NF-kappa B) activity were measured. RESULTS: M phi, PGE(2) and IL-6 production in Trauma+Intact mice was significantly increased compared with Control+Intact mice. Adrenalectomy decreased these levels to Control levels. Similar changes were observed for COX-2 and IL-6 expression. M phi nuclear NF kappa B levels were increased in Trauma+Intact mice compared with controls. Adrenalectomy abrogated this increase. Treating Trauma+Intact mice with RU-486 did not restore PGE(2) and IL-6 production or COX-2 and IL-6 messenger RNA to control levels. Administering exogenous glucocorticoid to Intact mice did not increase PGE(2) and IL-6 production or COX-2 and IL-6 mRNA to Trauma levels. CONCLUSIONS: The neuroendocrine system upregulates certain M phi inflammatory mediators, including PGE(2), IL-6, and NF-kappa B, after trauma. This upregulation does not seem to be mediated via glucocorticoids and possibly may be mediated via catecholamines. Elucidation of the interactions between the neuroendocrine system, the immune system, and inflammatory mediator secretion might provide novel therapeutic strategies for the injured patient. PMID- 15523399 TI - Effect of smoking and abstention on oxidative burst and reactivity of neutrophils and monocytes. AB - BACKGROUND: Smoking is associated with surgical wound infections, impaired wound healing, and tissue-destructive disorders. The mechanisms are largely unknown, but changes in the function and activity of inflammatory cells may be involved. METHODS: Seventy healthy volunteers (54 smokers and 16 never smokers) were included. The smokers were studied while they smoked and after 20 days of abstinence. After the first 10 days of abstinence, they were randomized to double blind treatment with transdermal nicotine patch 25 mg per day or placebo. Venous blood neutrophils and monocytes were sampled and isolated. In 22 randomly selected smokers and in all never smokers, the oxidative burst and chemotaxis were determined by a chemiluminescence response assay and a modified Boyden chamber technique, respectively. Stimulants were opsonized zymosan, formyl-Met Leu-Phe, and zymosan-activated serum. RESULTS: The neutrophil and monocyte oxidative burst was 50% and 68% lower, respectively, in smokers compared to never smokers (P < .05). Neutrophil chemotaxis was 93% higher in smokers (P < .05). Monocyte chemotaxis was lower in smokers compared to never smokers (P < .05). After 20 days of abstinence, neutrophil oxidative burst increased to the level of never smokers (P < .05); monocyte oxidative burst increased by 50% (P < .05). Chemotaxis was only marginally affected. The changes induced by abstinence were less pronounced in the transdermal nicotine patch group compared to the placebo group. CONCLUSIONS: Smoking attenuates the oxidative burst of inflammatory cells and increases chemotaxis. Three weeks of abstinence normalize the oxidative burst, but affect chemotaxis only marginally. PMID- 15523401 TI - Angiotensin II type 1 receptor activation increases microvascular hydraulic permeability. AB - BACKGROUND: In addition to its vasoconstricting effects, angiotensin II (Ang II) has also demonstrated the ability to modulate microvessel permeability. We hypothesized that activation of the angiotensin II type 1 receptor (AT1) would increase hydraulic permeability. METHODS: Hydraulic permeability (L(p)) was measured in rat mesenteric venules using the Landis micro-occlusion technique. Paired measures of L(p) were obtained at baseline and after perfusion with the AT1 agonist, [Sar(1)]-angiotensin II, at 10 micromol/L (n=6) and 100 micromol/L (n=6). Activation of the AT1 receptor was also achieved by perfusion with 20 nmol/L Ang II plus the angiotensin II type 2 receptor (AT2) antagonist, PD123319. In these studies, 30 micromol/L (n=6) and 300 micromol/L (n=6) of PD123319 were used. RESULTS: [Sar(1)]-angiotensin II increased L(p) 2-fold with the 10 micromol/L dose (P=.04) and 4-fold with the 100 micromol/L dose (P < .001). The L(p) peak due to [Sar(1)]-angiotensin II occurred sooner than the peak observed with Ang II. PD123319 (30 micromol/L) plus 20 nmol/L Ang II increased L(p) 5-fold (P=.003), while PD123319 (300 micromol/L) plus 20 nmol/L Ang II increased L(p) 20 fold (P < .0001). The magnitude of the effect due to PD123319 (300 micromol/L) plus Ang II (20 nmol/L) was approximately twice the summation of effects due to PD123319 (300 micromol/L) alone and Ang II (20 nmol/L) alone. CONCLUSIONS: We conclude that endothelial cell Ang II receptors play an important role in modulating transendothelial fluid flux. Activating the AT1 receptor increases L(p); the AT2 receptor may operate to oppose this action. Pharmacologic manipulation of Ang II receptors may be beneficial during shock states to limit intravascular fluid loss. PMID- 15523402 TI - Association between ductal fluid proteomic expression profiles and the presence of lymph node metastases in women with breast cancer. AB - BACKGROUND: Proteomic analysis of nipple aspiration fluid (NAF) is a noninvasive method for studying the local biologic microenvironment of the breast ducts where carcinoma originates. METHODS: NAF samples from each breast of 23 women with stage I or II unilateral invasive breast carcinoma were collected, and protein expression was analyzed comprehensively by using protein arrays and time-of flight mass spectrometry. Blinded hierarchical clustering analysis was performed to identify potential associations between protein expression patterns and clinicopathologic factors. RESULTS: After analysis of all spectra, 463 distinct peaks in the mass range of 7 to 30 kD were identified in NAF samples. Blinded hierarchical clustering analysis of protein expression patterns demonstrated a conservation of these patterns between the breasts of individual patients (P=.0003 x 10(-12)). Hierarchical clustering revealed an association between protein expression patterns, and the presence and absolute number of axillary lymph nodes containing metastases (P=.038). CONCLUSIONS: Protein expression patterns are highly conserved between cancerous and noncancerous breasts in women with unilateral invasive breast cancer; unique expression patterns may be associated with extent of disease. High-throughput proteomic methods may reveal biologically relevant proteins involved in carcinogenesis and progression of disease. PMID- 15523403 TI - Developing measures of surgeons' equipoise to assess the feasibility of randomized controlled trials in vascular surgery. AB - BACKGROUND: Equipoise is defined medically as a state of genuine uncertainty about the relative benefits of alternative treatment options. This study investigated individual and collective equipoise among vascular surgeons for controversial clinical questions to assess the feasibility of conducting randomized controlled trials. METHODS: Vascular surgeons throughout Australia and New Zealand received a survey by mail. RESULTS: Vascular surgeons (n=146, 77% response fraction) were able to quantify the strength of their treatment preferences and did so differentially between clinical scenarios using a simple scale. Almost one quarter (24%; 95% CI, 18%-32%) were completely undecided about whether carotid endarterectomy or carotid stenting was preferable to treat carotid stenosis in high-risk patients, indicating individual equipoise. In contrast, the vast majority of respondents (89%; 95% CI, 82%-93%) favored carotid endarterectomy over carotid stenting for average-risk patients, suggesting lack of community equipoise for this patient group. Similarly, there was lack of community equipoise for treatments for abdominal aortic aneurysm in high-risk patients with 88% (95% CI, 81%-92%) favoring a minimally invasive approach. Older respondents were consistently less willing to take part in randomized trials, with strength of treatment preference also independently predicting willingness to participate in 4 of 6 trials. CONCLUSIONS: Individual and community equipoise can be measured in a representative sample of surgeons as part of the feasibility assessment for future randomized controlled trials. PMID- 15523404 TI - Feasibility of breast duct lavage performed by a physician extender. AB - BACKGROUND: The safety and feasibility of ductal lavage (DL), a risk-assessment tool utilizing a minimally invasive technique that permits sampling of breast duct epithelium, performed primarily by a nurse practitioner (NP), was studied prospectively. METHODS: Women at high risk for breast cancer with a normal clinical breast exam and mammogram were enrolled. Nipple aspirate fluid (NAF) yielding ducts were identified, cannulated, and lavaged primarily by an NP in collaboration with a breast surgeon. Samples with sufficient cellularity were categorized as benign, mild atypia, marked atypia, or malignant. Pain and adverse events were recorded. RESULTS: Thirty-seven women, with a mean age of 51.7 years, were enrolled. Thirty-one (83.8%) women yielded NAF and, of those, 28 (90.3%) had one or more ducts successfully cannulated. Of 65 lavaged ducts in these 28 women, cellularity was adequate for diagnosis in 44 (67.7%) samples. Cytologic findings were as follows: 24 benign, 15 mild atypia, 4 marked atypia, and 1 malignant. The procedure was well tolerated with a mean pain score of 3.2 (SD +/- 1.81). The most frequent adverse event was breast fullness, reported by 44.8% of the women. Two women with marked atypia were evaluated further and found to have intraductal papillomata. The woman with malignant cytology had ductal carcinoma in situ. CONCLUSION: DL is a safe, generally well-tolerated procedure that can be performed successfully by a trained NP. PMID- 15523405 TI - Emergence of medullary thyroid carcinoma in a family with the Cys630Arg RET germline mutation. AB - BACKGROUND: Individual germline mutations in the RET (REarranged during Transfection) proto-oncogene may set the time window for malignant progression from C-cell hyperplasia to familial medullary thyroid carcinoma. Owing to the close genotype-phenotype correlation, genetic information may lend to individual timing of prophylactic thyroidectomy according to RET genotype. Limited information exists on the Cys630 RET genotype. Most of the few published carriers of this genotype who developed medullary thyroid carcinomas (MTCs) were in their mid-30s. METHODS: This case series of a German RET family with the Cys630Arg genotype was assembled to study malignant progression of MTC in this rare RET genotype. RESULTS: There was considerable variability of malignant progression from C-cell hyperplasia to MTC in carriers of the Cys630Arg genotype. In these persons, MTCs had developed by the age of 32 years (index patient, pT2bN0M0), and 15 years and 1 year (non-index patients; pT1apN1bM0 and pT1bpN0M0, respectively). The Cys630Arg genotype always segregated with the familial medullary thyroid carcinoma phenotype. CONCLUSIONS: The Cys630 RET genotype may have a more vigorous transforming activity than currently thought and can cause MTC in RET gene carriers within the first year of life. Starting in early infancy, identified RET gene carriers should be scrutinized until stimulated serum calcitonin levels become positive or, when these remain normal, should undergo prophylactic thyroidectomy before they reach 5 years of age. PMID- 15523406 TI - Combined modality treatment of ruptured pancreaticoduodenal artery aneurysms due to celiac artery compression. PMID- 15523407 TI - The military physician's ethical response to evidence of torture. PMID- 15523408 TI - Hopkins long ago (II). PMID- 15523409 TI - Mucocele of the appendix. PMID- 15523410 TI - Laparoscopic excision of bilateral primary adrenal leiomyosarcomas in a 14-year old girl with acquired immunodeficiency syndrome (AIDS). PMID- 15523411 TI - Re: "Thorough intraoperative analysis of breast sentinel lymph node biopsies: Histologic and immunohistochemical findings". PMID- 15523412 TI - Re: "The immunophenotype and activation status of the lymphocytic infiltrate in human breast cancers, the role of the major histocompatibility complex in cell mediated immune mechanisms, and their association with prognostic indicators". PMID- 15523415 TI - Cell phones: must they be used all the time? PMID- 15523416 TI - Spiritual development and the dying child: the pediatric nurse practitioner's role. AB - Dying children require special consideration when it comes to spiritual development. Pediatric nurse practitioners (PNPs) can be a pivotal influence in the manner in which a child progresses from sick child to dying child and an influence on the family at large during the process. The primary care PNP is uniquely positioned to have a premorbid knowledge of the spiritual development and needs of the child and family. The tertiary care PNP has a responsibility to provide continuing spiritual assessment and follow-up with referrals to appropriate professional staff and community resources for spiritual care. Childhood spiritual development within the context of the PNP's role in assessing and meeting the spiritual needs of the dying child is discussed. PMID- 15523417 TI - Achieving continence in children with neurogenic bowel and bladder. AB - Bowel and bladder continence is a major developmental accomplishment for a child and his or her family. Children with neurogenic bowel and bladder dysfunction, which occurs with myelomeningocele and other neural tube defects, have a physical inability to attain continence. However, children with these limitations can attain continence when the appropriate modifications to the traditional routines are made. Enabling the child to attain continence improves parental relationships and self-concept. The nurse who is working with children with myelomeningocele in schools, clinics, private offices, and home health care settings can play a vital role in helping a child with a neural tube defect achieve continence. PMID- 15523418 TI - A qualitative study of older first-time mothering in the first year. AB - INTRODUCTION: The number of older first-time mothers has increased significantly during the past 30 years. The purpose of this study was to create a phenomenologic text explicating the experience of older, first-time mothering during the first year. METHOD: A phenomenologic/hermeneutic method was used to create a narrative text to address the following question: In what way does older age give a certain quality of meaning to first-time motherhood in women 35 years of age and older? This text incorporated data from interviews with seven first time mothers ages 36 to 48 years, as well other data sources such as poetry, literary references, and findings from other qualitative studies. RESULTS: Among essential themes that emerged from analysis of study data were: "Being Ready," "Planned Intensity," and "How Far We've Come." DISCUSSION: Pediatric practitioners have unparalleled access to mothers during the first postpartum year. An increased understanding of the uniqueness of older primiparas as expressed in this narrative will enable nurses to more effectively address the needs and concerns of this growing population of women. PMID- 15523419 TI - Facts and fallacies: the practice doctorate. AB - In the past year there has been continued discussion within professional organizations regarding the emergence of the practice doctorate. This in turn has caused concerns among some nurse practitioners. The purpose of this article is to provide an overview of the practice doctorate, current progress and development of the concept, and the potential impact on practicing pediatric and other nurse practitioners. PMID- 15523420 TI - Viral croup: a current perspective. AB - Viral croup is the most common cause of upper airway obstruction in children 6 months to 6 years of age. Parainfluenza virus accounts for the majority of cases. The disease is characterized by varying degrees of inspiratory stridor, barking cough, and hoarseness because of laryngeal and/or tracheal obstruction. The diagnosis is mainly a clinical one and diagnostic studies usually are not necessary. The management has altered dramatically in the past decade. Good evidence exists to support the routine use of corticosteroid in all children with croup. Intervention at an earlier phase of the illness will reduce the severity of the symptoms and the rates of return to a health care practitioner for additional medical attention, visits to the emergency department, and admission to the hospital. Most children respond to a single, oral dose of dexamethasone. For those who do not tolerate the oral preparation, nebulized budesonide or intramuscular dexamethasone are reasonable alternatives. Nebulized epinephrine should be reserved for patients with moderate to severe croup. Simultaneous administration of corticosteroid and epinephrine reduces the rate of intubation in patients with severe croup and impending respiratory failure. PMID- 15523422 TI - Grief masks. PMID- 15523421 TI - Comparison of children's fears of medical experiences across two cultures. AB - INTRODUCTION: This study examined the self-reported fears of school-age children living in the United States and in Nepal. METHOD: Thirty school-age children from each country, matched by sex and age, participated in this study. The revised version of the Child Medical Fear Scale was used to identify medical fears of children. RESULTS: Among 17 feared items, the result showed getting a shot to be the most feared item reported by both groups. Nepalese children reported higher fear scores than did American children. It also was found that there was a significant difference in fear scores between Nepalese boys and girls, with girls reporting higher fear scores than boys. There was no difference in fear scores between American boys and girls. DISCUSSION: The findings of this study are important to nurse practitioners in understanding children's fears of medical experiences across different cultures. Nurses can support children in dealing with their fears of medical experiences by recognizing these fears and taking into consideration the child's family, sex, and culture when planning care. Nursing implications are discussed. PMID- 15523423 TI - Abdominal pain with stripes. PMID- 15523424 TI - Child care: a crucial legislative issue. PMID- 15523425 TI - Preventing childhood obesity: establishing healthy lifestyle habits in the preschool years. PMID- 15523426 TI - Constipation: an overview of treatment. PMID- 15523427 TI - The effects of superoxide dismutase in gerbils with bacterial meningitis. AB - BACKGROUND: Inflammatory products, such as oxygen radicals generated during the course of bacterial meningitis, can damage nerve endings, hair cells, and/or supporting cells in the cochlea. Superoxide dismutase (SOD), an O2-scavenger, has been shown to play an important role in the protection against radical toxicity in various animal experiments. OBJECTIVE: To study the antioxidant effects of SOD on the inflammatory response of gerbils with bacterial meningitis. STUDY DESIGN: Meningitis was induced in three groups of 10 gerbils by intrathecal (IT) injection of Streptococcus pneumoniae into the cisterna magna. Group 1 received IT SOD, group 2 received intramuscular (IM) SOD, and group 3, the control group, received IM normal saline. Histologic data and auditory brainstem responses (ABR) were obtained from each gerbil. RESULTS: Fibrosis and/or neo-ossification were near absent in the IT SOD group and significantly less fibrosis occurred in the IM group (IT vs. IM: P = 0.010; IT vs. control group: P = 0.001). The amount of surviving spiral ganglion cells correlated inversely with the extent of fibrosis (r = -0.753, P < 0.00001). CONCLUSIONS: IT injection of SOD significantly reduced cochlear fibrosis and neo-ossification, reduced the spiral ganglion cell loss, and decreased damage of the cochlear components following bacterial meningitis. PMID- 15523429 TI - Phenotypic and microarray gene expression analysis of tri-dimensional raft modeled human head and neck squamous cell carcinoma. AB - OBJECTIVES: To describe the phenotypic and gene expression differences in monolayer and tri-dimensional cultures systems. METHODS: Normal oral epithelial cells (NOEC), primary head and neck squamous cell carcinoma (HNSCC), and HNSCC cell lines were used to create and study modeled tri-dimensional tissue. Using cDNA microarray analysis, monolayer and raft-modeled tri-dimensional HNSCC cell lines were compared. RESULTS: NOEC, HNSCC, and both together can be modeled with tri-dimensional differentiation and cytokeratin characteristics analogous to in vivo tissue. Modeling of primary HNSCC resulted in a morphology of invasive HNSCC with areas of direct collagen invasion and MMP2 expression. Gene array analysis suggests that the individual cell lines themselves are the primary gene expression predictor and not the presence of tri-dimensional tissue architecture. CONCLUSIONS: This tissue culture modeling system approximates the differentiation and tri-dimensional structure of in vivo tissues, and that raft modeled tri dimensional HNSCC does not have a significantly different gene expression profile than the corresponding monolayer culture. PMID- 15523428 TI - Modulation of Th1 and Th2 cytokine profiles and their association with advanced head and neck squamous cell carcinoma. AB - OBJECTIVE: Plasma cytokine concentrations from patients with head and neck squamous cell carcinoma (HNSCC) were measured to determine whether the potential modulation of host Th1 vs Th2 immune responses are associated with advanced clinical disease. STUDY DESIGN AND SETTING: The concentrations of IL-4, IL-6, IL 10, and IL-12 were measured in the plasma of 58 patients with histologically proven HNSCC. These data were examined with respect to the histologic size (T stage) of the primary tumor, and presence of nodal metastasis. RESULTS: The concentrations of IL-12 were greater from patients without nodal metastasis, and with T(1)/T(2)-stage tumors. IL-10 levels were greater from patients with nodal metastasis, and with T(3)/T(4)-stage tumors. The concentrations of IL-6 were greater from patients with T(3)/T(4)-stage tumors. CONCLUSIONS: Using parameters of primary tumor size and presence of nodal metastasis, patients with advanced HNSCC have significantly less plasma IL-12 levels, and greater plasma IL-10 and IL-6 levels. SIGNIFICANCE: Patients with advanced HNSCC have a potentially diminished Th1 immune response, and a stronger potential Th2 immune response when compared to that of patients with less advanced disease. EBM RATING: D-5. PMID- 15523430 TI - Alterations in eotaxin, monocyte chemoattractant protein-4, interleukin-5, and interleukin-13 after systemic steroid treatment for nasal polyps. AB - OBJECTIVE: To determine alterations in Th2 chemokines eotaxin and monocyte chemoattractant protein-4 (MCP-4), and cytokines interleukin-5 (IL-5) and interleukin-13 (IL-13), in nasal polyps (NP) after steroid treatment. STUDY DESIGN: Cytokine/chemokine levels were measured in NP before and after steroid therapy and compared to control sinus mucosa. RESULTS: Twenty-one patients (control = 7, NP = 14) were enrolled. Eotaxin and MCP-4 were significantly higher than control tissue (P = 0.004 and 0.003). All four mediators decreased after steroid treatment (P < 0.03). IL-5 and IL-13 in untreated polyps were not significantly different from controls. Patients showed clinical improvement according to SNOT-20 scores (average presteroid score 19, poststeroid score 13) and endoscopic grading (1.75 each side presteroid, 1.13 poststeroid). CONCLUSIONS: Steroids significantly decreased all cytokine/chemokine levels, but the impact on Th2 chemokines was of a much greater magnitude. SIGNIFICANCE: Novel approaches to block inflammatory mediators, particularly Th2 chemokines, may lead to better control of nasal polyposis in the future. PMID- 15523431 TI - Selective acquisition of individual cell types in the vestibular periphery for molecular biology studies. AB - OBJECTIVES: To develop a method for characterizing the transcriptome of individual cell types in the inner ear sensory epithelia. STUDY DESIGN: We employed the technique of laser capture microdissection to obtain enriched populations of hair cells and supporting cells. The respective mRNAs were extracted, reverse transcribed, and amplified using PCR. RESULTS: We were able to isolate RNAs with good integrity from enriched cell populations obtained with laser capture microscopy and amplify specific mRNA targets. CONCLUSIONS: We can now investigate the molecular differences between the different cell types in the inner ear sensory epithelia as identified by morphological criteria. SIGNIFICANCE: Analysis of gene expression profiles in the inner ear cell types has been hampered by the small size of this tissue and by the compact histoarchitecture of the sensory epithelia; however, the present technique offers new possibilities for the analysis of transcriptomes in the vestibular periphery using available high-throughput gene expression analysis methods. PMID- 15523432 TI - Continuous laryngeal nerve integrity monitoring during thyroidectomy: does it reduce risk of injury? AB - OBJECTIVE: To determine if continuous laryngeal nerve integrity monitoring (NIM) during thyroidectomy is associated with a decreased risk of postoperative recurrent laryngeal nerve (RLN) injury. STUDY DESIGN AND SETTING: Retrospective cohort study of 165 patients who underwent thyroidectomy at a resident teaching institution between 1999 and 2002. The control group had 120 nerves at risk (NAR) whereas the NIM group consisted of 116 NAR. Primary outcome measures included postoperative RLN paralysis, paresis, and total injury rates. RESULTS: RLN paralysis occurred in 2.54% NAR in the control group and 0.86% in the NIM group [relative risk (RR) = 0.34, 95% confidence interval (CI95) = 0.04-3.27, P = 0.62]. Temporary RLN paresis occurred in 4.24% NAR in the control group and 3.45% in the NIM group (RR = 0.9, CI95 = 0.23-3.55, P = 0.89). Total RLN injury occurred in 6.78% NAR in the control group and 4.31% in the NIM group (RR = 0.66, CI95 = 0.21-2.09, P = 0.48). When compared to all other NAR, advanced T-stage malignancy was associated with a significantly increased risk of RLN paresis (T4 RR = 9.0, CI95 = 2.56-31.67, P = 0.0006; T3+T4 RR = 7.5, CI95 = 2.17-25.86, P = 0.0001) but not paralysis. NIM did not significantly reduce the risk of RLN paresis in the advanced T-stage subset (T3+T4 RR = 0.36, CI95 = 0.04-3.0, P = 0.59). CONCLUSION: There were no statistically significant differences in RLN paralysis, paresis, or total injury rates between control and NIM groups, even in subsets with advanced T-stage and increased baseline risk. Advanced T-stage is a significant predictor of RLN paresis in this cohort. PMID- 15523434 TI - Study of lubricant-induced changes in chronic snorers (SLICCS). AB - OBJECTIVE: The efficacy of many of the noninvasive treatments for snoring has not been evaluated in controlled trials. This paper seeks to evaluate the efficacy of an oil-based spray in the treatment of snoring, in a double-blinded, placebo controlled, crossover trial using objective acoustic analysis and subjective questionnaires. STUDY DESIGN AND SETTING: Participants were randomized to use both oil-based oral spray (treatment) and water-based oral spray (placebo) during a two-night in-home study period. Questionnaires were completed by participant and bed-partner in addition to audio-tape recordings which were analyzed for frequency, duration, and mean energy of snoring. RESULTS: Greatest snoring rate demonstrated 30% = benefit; 40% = no change; 30% = adverse effect (n = 20). Percent time snoring yielded: 30% benefit; 15% no change; 55% adverse effect (n = 20). Study data results for mean energy were (n = 12): benefit = 17%, no change = 33%, adverse effect = 50%. Bed-partner observations (n = 17) demonstrated 37% = benefit; 38% = no change; 25% = adverse effect. CONCLUSION/SIGNIFICANCE: Objective and subjective evaluation of the performance of the oil-based Snoreless spray in comparison to placebo demonstrated a lack of efficacy in snoring reduction. PMID- 15523433 TI - Prospective, randomized, controlled clinical trial of a novel matrix hemostatic sealant in children undergoing adenoidectomy. AB - PROBLEM ADDRESSED: Floseal is a novel matrix hemostatic sealant composed of collagen-derived particles and topical bovine-derived thrombin. It is applied as a high-viscosity gel for hemostasis and has been clinically proven to control bleeding. This study is a prospective, randomized, controlled clinical trial of Floseal sealant compared to traditional suction cautery hemostasis in children undergoing adenoidectomy. METHODS AND MEASURES: Seventy patients (mean age 7.0 yrs, 45.7% male) with obstructive sleep apnea underwent traditional cold steel adenoidectomy with an adenoid curette and were then randomized to receive the hemostatic sealant (Floseal) or cautery to obtain hemostasis. Patients were crossed over to the other hemostatic technique if hemostasis was not achieved after more than 100 mL of blood loss or 15 minutes elapsed time. Objective data collected included time to hemostasis and blood loss during hemostasis. Visual analog scales (VAS) were used to record subjective data by the operating surgeon including bleeding following adenoid pack removal (0 = none, 3 = brisk) and ease of operation (1 = extremely easy, 6 = extremely difficult). Parents recorded diet on a journal and were contacted by phone at postoperative day 7 and questioned with regard to return to regular diet and use of narcotics. RESULTS: Compared to patients in the cautery group (n = 35), Floseal patients (n = 35) had significantly shorter times to hemostasis (0.6 +/- 1.3 minutes vs 9.5 +/- 5.4 minutes [mean +/- SD], P < 0.001), less blood loss (2.5 +/- 9.2 mL vs 29.4 +/- 27.1 mL, P < 0.001), less subjective bleeding (0.0 +/- 0.6 vs 2.0 +/- 0.7, [median 4-point VAS +/- SD], P < 0.001), and subjectively easier operations (2.6 +/- 1.0 vs 5.2 +/- 1.0 [mean 6-point VAS +/- SD], P < 0.001). Furthermore, Floseal patients returned to regular diet earlier (2.7 +/- 0.7 vs 4.1 +/- 0.5 days [mean +/- SD], P < 0.001) and had less use of narcotics at 7 days postoperatively (40% vs 69%, P < 0.05). Lastly, three patients in the cautery group were crossed over to the Floseal group, but no Floseal subjects were crossed over to the cautery group. The retail cost of Floseal is US 85 dollars. Operating room costs are estimated at US 12 dollars/minute. Reducing the operative length by 8.9 minutes on average produces a cost savings of US 106.80 dollars per operation. There were no complications in either experimental group including postoperative hemorrhage, hospitalization, blood transfusion, or aspiration. CONCLUSIONS: Floseal matrix hemostatic sealant is a safe, efficacious, easy, and cost-effective technique for obtaining hemostasis in children undergoing adenoidectomy. Limitations of the study include the fact that it is nonblinded, which does allow for some bias in the subjective data recorded. However, utilizing 4 different operating surgeons, 3 of whom were not affiliated with the study, minimized this. CLINICAL SIGNIFICANCE OF STUDY: This study demonstrates the safety and efficacy of a novel hemostatic sealant in children undergoing adenoidectomy. Floseal matrix hemostatic sealant can be used as a first-line hemostatic agent, and it is a good tool in the armamentarium of otolaryngologists who encounter significant bleeding following adenoidectomy. PMID- 15523435 TI - The dilemma of the normal baseline parathyroid hormone level using the intraoperative PTH assay. AB - OBJECTIVE: To analyze patients with "normal" baseline quick intraoperative parathyroid hormone (QPTH) levels during parathyroidectomy and to determine the prevalence of this finding, the usefulness of the assay in this situation, and to explain the possible causes for this phenomenon. STUDY DESIGN AND SETTING: Patients who underwent parathyroidectomy using QPTH in a tertiary hospital. METHODS: Retrospective analysis of 39 patients treated surgically for primary hyperparathyroidism using QPTH. RESULTS: Of the patients, 14 (36%) had normal baseline QPTH. 8 patients with localizing sestamibi scans had a single adenoma, and excision resulted in a mean decrease of 85.4% in QPTH. Six patients had nonlocalizing sestamibi scans, 1 patient had an 84% drop in QPTH level after removal of a single adenoma, and 5 patients had hyperplasia requiring > or =3 glands excision. At 11.36 months' mean follow-up, 13 patients (93%) were normocalcemic. CONCLUSIONS: A "normal" baseline QPTH level was found in 36% of patients. A 50% decrease in QPTH remains predictive of biochemical cures in patients with localizing sestamibi scans. The likely explanation for this variability in "normal" levels between different assays is the variability in detection of the 7-84 PTH fragment, which results in an overestimation of the PTH level. Assays such as the QPTH, which are more sensitive for the biologically active PTH molecule [(1-84) PTH] than other laboratory PTH assays will tend to have lower PTH levels that can be within the normal range. EBM RATING: B-3. PMID- 15523436 TI - Rapid intraoperative localization of parathyroid glands utilizing methylene blue infusion. AB - OBJECTIVE: To review a single surgeon's experience utilizing an intraoperative methylene blue infusion (IMBI) to identify parathyroid glands during neck exploration for primary hyperparathyroidism. STUDY DESIGN AND SETTING: Retrospective review of 35 patients who underwent bilateral neck exploration utilizing an IMBI at a dose of 7.5 mg/kg following the induction of general anesthesia. RESULTS: All patients reverted to normocalcemia with a mean follow-up of 17 months. IMBI facilitated the identification of abnormal parathyroid tissue in 34/35 patients (97%). A dark blue-purple staining was observed in 33/37 stained adenomas (89%). Four adenomas and four hyperplastic glands stained a lighter shade of blue-green. Among 89 normal glands, 41(46%) stained a pale green grey color. CONCLUSIONS: IMBI is a safe, readily available, cost-effective, and underutilized technique that facilitates rapid identification of parathyroid adenomas, helps distinguish normal glands from hyperplastic glands, and helps to locate ectopic glands. An overall reduction in operative time, especially for bilateral neck exploration, can be anticipated. PMID- 15523437 TI - The fluorescence of thyroid tissue. AB - OBJECTIVE: To investigate the fluorescent characteristics of different thyroid tissues. STUDY DESIGN: Fresh ex vivo thyroid tissue was surfaced scanned with a fluorescence spectrophotometer. Resultant spectra were evaluated via interpeak ratios. RESULTS: Interpeak ratios of all histologies were found to be significantly different from each other (P < 0.001). The diagnostic sensitivity and specificity of fluorescent spectroscopy was 86% and 91% for thyroid nodule capsule, 78% and 59% for papillary thyroid carcinoma, and 82% and 63% for normal thyroid, respectively. CONCLUSION: Fluorescent spectroscopy can been used to identify significant differences in the fluorescent characteristics of various thyroid histologies. SIGNIFICANCE: This study is the first investigation of the fluorescence of thyroid tissue. The diagnostic ability of fluorescent spectrometry may allow it to be used as a localization aid for fine needle aspiration, using optical fiber probes. Further investigations may enhance the sensitivity and specificity of fluorescent spectrometry allowing it to replace or compliment fine needle aspiration. EBM RATING: B-3. PMID- 15523438 TI - Primary hyperparathyroidism in young adults. AB - OBJECTIVES: The purpose of this study was to compare the incidence of multiglandular disease and rate of treatment failure between younger and older patients with primary hyperparathyroidism. STUDY DESIGN AND SETTING: The medical charts of patients who underwent surgery for primary hyperparathyroidism at our tertiary-care institution between 1995 and 2001 were reviewed. RESULTS: Three hundred nineteen patients were identified, of whom 33 were aged 40 years or less. There were no statistically significant differences between the younger and older groups in the incidence of multiglandular disease (9.1% for both, P = 1.00) or in the treatment failure rate (12.1% and 8%, respectively, P = 0.43). Sonography was significantly more sensitive than technetium Tc-sestamibi in the younger group (96% vs 57%, P < 0.05). Parathyroid hormone level and gland weight were significantly higher in the older group (P = 0.004). CONCLUSION: Our results suggest that the same treatment strategy should be applied to all patients with primary hyperparathyroidism. Ultrasound appears to be the localization procedure of choice in younger patients. PMID- 15523440 TI - Contemporary staging and prognosis for primary tracheal malignancies: a population-based analysis. AB - OBJECTIVE: Determine staging characteristics and survival outcomes for primary malignancies of the trachea. Design Cross-sectional analysis of national cancer database. METHODS: Cases of primary tracheal malignancy were extracted from the Surveillance, Epidemiology, and End Results database for the time period 1988 2000. T-stage, N-stage, and overall stage of presentation were determined. Mean, median, and 5-year survival statistics were computed using Kaplan-Meier survival analysis for each tumor histology and for the overall cohort according to stage. RESULTS: Ninety-two cases with adequate histologic information were identified. Mean age at presentation was 59.3 years with an equal sex distribution. Squamous cell carcinoma was the most common tumor type (41 cases) followed by adenoid cystic carcinoma (19 cases). Forty-nine cases (53%) presented with stage 3 or stage 4 disease. Squamous cell carcinoma exhibited poorer survival (mean survival, 44.0 month, 5-year survival, 34%) than adenoid cystic carcinoma (mean survival, 115 month, 5-year survival, 78%). Five-year unadjusted survival rates according to overall stage were 52.8%, 70.0%, 75.0%, 15.1%, respectively. CONCLUSIONS: Primary tracheal malignancies often present with advanced stage. Patients with squamous cell carcinoma of the trachea have poorer prognoses when compared with adenoid cystic carcinoma and other tumor types. Staging tracheal cancer with a TNM-based system helps predict survival. EBM RATING: C. PMID- 15523439 TI - Pseudoaneuploid subpopulations detected in normal upper aerodigestive tract mucosa consistent with physiological apoptosis in normally differentiating squamous mucosa. AB - OBJECTIVES: While evaluating the validity of using normal human mucosal cells from the upper aerodigestive tract as diploid standards for DNA content studies of squamous cell cancer of head and neck by flow cytometry, pseudoaneuploidy was frequently detected. The purpose of this study was to further evaluate these DNA content abnormalities encountered in normal human mucosal cells and correlate them to physiological apoptosis. STUDY DESIGN: Thirty-two specimens of upper areodigestive tract mucosa from 18 surgical resections, 11 fresh autopsies, and 3 buccal scrapings were examined for DNA content by flow cytometry. RESULTS: Pseudoaneuploidy, which ranged from sub-G0/G1 peaks to hyperdiploid peaks with increased 90 degrees light scattering properties was found in 60% of these specimens. Fluorescent microscopic examination of the sorted DNA pseudoaneuploid cells demonstrated cells undergoing apoptosis. CONCLUSION: This unexpected pseudoaneuploidy in normal mucosal cells was a result of physiological apoptosis, a normal component of squamous differentiation. EBM RATING: B-2. PMID- 15523441 TI - Radiation-induced malignancy of the head and neck. AB - OBJECTIVE: Our goal was to evaluate our experience with radiation-induced malignancy (RIM), compare that experience to the literature, and review treatment modalities. STUDY DESIGN AND SETTING: The setting is the University of Kansas Medical Center. A retrospective review was performed to identify patients with RIM. Patients were included if they met the criteria for RIM as delineated in the literature. RESULTS: Thirteen patients met the criteria for RIM. The mean latency period was 22 years. Sarcomas were the most common type of RIM and the paranasal sinuses were the most common location. Surgical resection was our treatment of choice. CONCLUSIONS: Our patient series differs from previous reports in that sarcomas were the predominating RIM and the paranasal sinuses were the most common location. We noted a shorter latency period than has been previously published. Surgical excision is the treatment of choice. EBM RATING: C. PMID- 15523442 TI - Correlation of histopathological variants, cellular DNA content, and clinical outcome in adenoid cystic carcinoma of the salivary glands. AB - OBJECTIVE: To study the correlation between flow cytometrically measured DNA ploidy with prognostically important histopathologic groups and clinical outcome in patients with adenoid cystic carcinoma of the salivary glands. STUDY DESIGN: 46 tumor specimens were analyzed flow cytometrically for DNA content and assessed for histological grade. Correlations were made between tumor DNA ploidy and histopathological grade, and disease-free and overall survival of these patients. RESULTS: Of the 46 patients, 31 had a cribiform/tubular histologic pattern, and 15 had a solid pattern. 84% of the tumors with cribriform/tubular pattern were DNA diploid, compared with 33% of tumors that were graded solid. This difference proved to be statistically significant (chi(2)11.75, P = 0.0006). Overall and disease-free survival periods were longer for patients with DNA diploid tumors in both groups, 63% vs. 36% and 62% vs 38%, respectively. CONCLUSIONS: Tumor DNA ploidy correlates with prognostically important tumor histopathology as well as overall and disease-free survival in patients with adenoid cystic carcinoma of the salivary gland. EBM RATING: B-3. PMID- 15523443 TI - Molecular diagnosis of nasopharyngeal carcinoma: detecting LMP-1 and EBNA by nasopharyngeal swab. AB - OBJECTIVES: The aim of this study was to investigate the feasibility of molecular diagnosis of nasopharyngeal carcinoma (NPC) by combining nasopharyngeal swab and polymerase chain reaction (PCR) to detect Epstein-Barr virus (EBV) derived latent membrane protein-1 gene (LMP-1) and Epstein-Barr nuclear antigen gene (EBNA). METHODS: 437 adults underwent nasopharyngoscopy, possible biopsy, and nasopharyngeal swab to obtain nasopharyngeal cells for the detection of LMP-1 and EBNA by PCR. RESULTS: By detecting LMP-1 and EBNA in the nasopharyngeal swabs, NPC could be diagnosed with a false-positive rate of 1.7% (6/354), a false negative rate of 8.6% (6/70), a sensitivity of 91.4% (64/70), a specificity of 98.3% (348/354), positive predictive value of 91.4% (64/70), and negative predictive value of 98.3% (348/354). CONCLUSION: Detecting EBV genomic LMP-1 and EBNA by nasopharyngeal swab verifies NPC with a sensitivity of 91.4% and specificity of 98.3%. The nasopharyngeal swab coupled with PCR based EBV LMP-1 and EBNA detection could serve as a good supplement to pathological diagnosis of NPC. PMID- 15523444 TI - Selective neck dissection for clinically N0 neck in laryngeal cancer: is dissection of level IIb necessary? AB - The most common morbidity associated with selective neck dissection (SND; II-IV) is spinal accessory nerve dysfunction and related shoulder disability. Nerve dysfunction is usually attributed to stretching of the nerve during clearance of lymph nodes lying posterior and superior to the spinal accessory nerve (level IIb). If these lymph nodes were left in place and not removed, stretching of the spinal accessory nerve during neck dissection and postoperative shoulder disability could be avoided. 113 SNDs (II-IV) performed on clinically N0 necks of patients with laryngeal carcinoma were enrolled in this prospective study. During SND, level IIb was separately removed and processed. Mean number of lymph nodes in level IIb was 6.26 (range, 0-19). In none of the 113 SND (II-IV) specimens did level IIb contain metastases, thus providing an oncological basis that leaving these lymph nodes in place is an oncologically safe approach, probably avoiding postoperative shoulder disability. PMID- 15523445 TI - Use of the 2.0-mm locking reconstruction plate in primary oromandibular reconstruction after composite resection. AB - OBJECTIVE: To review our experience with 2.0-mm locking reconstruction plate (LRP) system for patients requiring oromandibular reconstruction. STUDY DESIGN: Retrospective case review of 43 consecutive patients who underwent mandibular composite resection with immediate reconstruction. SETTING: Tertiary care center. RESULTS: Forty-three patients underwent oromandibular reconstruction with the 2.0 mm mandibular LRP system and free flaps containing vascularized bone. Mean follow up was 11 months. There were no intraoperative difficulties utilizing this system. Two (5%) patients had partial fasciocutaneous flap loss resulting in plate exposure. There were no instances of plate fracture or complications requiring plate removal to date. CONCLUSION: 2.0-mm LRP mandibular system is reliable even in the setting of previous or adjuvant radiation therapy. Its technical ease of application, contouring malleability, and very low profile have proven to be advantageous in oromandibular reconstruction. SIGNIFICANCE: No previous descriptions of use of the 2.0-mm LRP in combination with osteocutaneous free flaps for mandibular reconstruction are found in the literature. EBM RATING: C. PMID- 15523446 TI - The impact of fiducial distribution on headset-based registration in image-guided sinus surgery. AB - OBJECTIVE: The objective of this study was to assess registration error due to fiducial configuration for the ENT headsets for the CBYON Suite (CBYON, Mountain View, CA) and InstaTrak (GEMS Navigation and Visualization, Waukesha, WI). STUDY DESIGN: Axial CT scans (1-mm slice thickness) were obtained of for 24 cadaveric heads using the CBYON headset and for 23 cadaveric heads using the GEMS headset. The CBYON and GEMS NAV software were used to calculate the fiducial registration error (FRE). Fiducial localization error (FLE) was estimated from FRE. Theoretical target registration error (TRE) was calculated at 11 targets. RESULTS: The FRE for CBYON and GEMS NAV was 0.69 mm and 0.27 mm, respectively. The theoretical TRE for CBYON and GEMS NAV was 0.41 mm and 0.30 mm, respectively. The theoretical TRE was greater at targets posterior in the sinus cavities. CONCLUSION: Theoretical TRE values for both ENT headsets are less than clinically observed TRE. Clinically observed TRE is likely due to repositioning accuracy. EBM RATING: B-2. PMID- 15523447 TI - Safety and efficacy of radioallergosorbent test-based allergen immunotherapy in treatment of perennial allergic rhinitis and asthma. AB - OBJECTIVE: This study was undertaken to demonstrate the safety and efficacy of in vitro, radioallergosorbent test (RAST)-based inhalant allergen immunotherapy. STUDY DESIGN AND SETTING: Prospective 22 year single site clinical study, with outcome evaluations of 480 perennial allergic rhinitis patients, including 96 with concomitant asthma. RESULTS: Rhinitis symptom control after 2 years of immunotherapy was excellent in 32.5% of patients, good in 45.6%, and fair in 14.2%. There was no improvement in 7.7%. For patients with asthma, 81% had good or excellent pulmonary symptom improvement, and no patient failed to improve. No severe reactions occurred, but there were 5 limited systemic reactions, or 0.008% of injections, during a 2.5-year mean immunotherapy treatment course. CONCLUSION: RAST-based immunotherapy is safe and effective for patients with perennial allergic rhinitis, with or without concomitant asthma. Significance This is the first large, multiyear study of safety and efficacy of RAST-based immunotherapy for treatment of perennial allergic rhinitis and asthma. EBM RATING: C. PMID- 15523448 TI - Effects of buffered saline solution on nasal mucociliary clearance and nasal airway patency. AB - OBJECTIVE: To compare the effects of buffered hypertonic and buffered normal saline nasal spray on mucociliary clearance and nasal airway patency. STUDY DESIGN AND SETTING: Double-blind trial with subjects acting as their own controls. Tertiary care academic medical center. RESULTS: Buffered hypertonic saline and buffered normal saline both improved saccharine clearance times ( P < 0.0001 for buffered hypertonic and P = 0.002 for buffered normal saline). Buffered hypertonic saline improved saccharine clearance times more than buffered normal saline (39.6% vs 24.1%, P = 0.007). Neither buffered hypertonic nor buffered normal saline significantly affected nasal airway patency. CONCLUSIONS: Both buffered hypertonic and buffered normal saline nasal spray significantly improved saccharine clearance times without affecting nasal airway patency. Buffered hypertonic saline affected saccharine clearance times to a greater degree than buffered normal saline. CLINICAL SIGNIFICANCE: Buffered hypertonic and buffered normal saline sprays both improve mucociliary clearance and should therefore be beneficial in conditions such as rhinitis and sinusitis, which are associated with disruption of mucociliary clearance. However, these sprays do not appear to affect the nasal airway. Patients may therefore benefit from other treatments for "nasal congestion." EBM RATING: B-2. PMID- 15523449 TI - Endoscopic approach to orbital blowout fracture repair. AB - OBJECTIVES: Transconjunctival and subciliary approaches to orbital floor blowout fractures (OBF) have known risks of postoperative eyelid malposition. This study evaluates the endoscopic transmaxillary repair of OBFs in a cadaveric model and clinical setting. METHODS: Sixteen cadaveric and 10 clinical OBFs were repaired endoscopically. A Caldwell-Luc approach and modified sinus surgery instrumentation were used to repair each fracture. Variations in fracture pattern, instrumentation, and surgical technique were evaluated. RESULTS: Endoscopic repair was achieved in all 16 cadaveric orbits and in 9 of 10 patients. Fracture patterns were classified as either medial or lateral to the infraorbital nerve. Average clinical operating time was 1:38. Of 9 postoperative CT scans, 5 were rated as excellent, 3 as good, and 1 as poor. CONCLUSIONS: The endoscopic transmaxillary approach is a safe, viable technique for OBF repair. It offers improved visualization, anatomic fracture repair, no risk of postoperative eyelid complications, and good clinical results. EBM RATING: C. PMID- 15523450 TI - Protein nitration in chronic sinusitis and nasal polyposis: role of eosinophils. AB - OBJECTIVES: To investigate the possible role of eosinophil peroxidase (EPO) in 3 nitrotyrosine (3NT) formation. STUDY DESIGN AND SETTING: Observational study employing immunocytochemistry to assess the presence of 3NT, inducible nitric oxide synthase (iNOS), eosinophils, mast cells, neutrophils, and lymphocytes in ethmoid sinus mucosal biopsies from normal controls and subjects with allergic and nonallergic chronic sinusitis and nasal polyposis. RESULTS: 3NT was more evident in biopsies from sinusitis patients (2.67 +/- 0.14, n = 21) than in healthy mucosa (0.43 +/- 0.2, n = 7, P < 0.01), but scores in atopic and nonatopic subjects were similar. Colocalization studies confirmed that 3NT was largely confined to eosinophils. No relationship was found between 3NT and other immune cells. 3NT detection was not correlated with the amount of immunostaining for iNOS. SIGNIFICANCE: Chronic sinusitis is accompanied by 3NT formation, which is largely restricted to the eosinophils, and likely driven by the action of eosinophil peroxidase, rather than by nitric oxide levels. EBM RATING: B-2. PMID- 15523451 TI - Allergic fungal sinusitis: can we predict the recurrence? AB - OBJECTIVE: The goal of the present study was to find out if recurrence can be predicted in cases of allergic fungal sinusitis. We also studied the influence of postoperative corticosteroid therapy on recurrence following surgery. STUDY DESIGN AND SETTING: This study was conducted at the ENT Department of Al Nahdha Hospital, which is a tertiary referral and teaching hospital in Muscat, Sultanate of Oman. The study is a retrospective analysis of 32 cases of allergic fungal sinusitis. Age, sex, extent of disease, and preoperative serum IgE levels were compared in patients who had recurrence with those who did not. We also studied the incidence, onset, and severity of recurrence in patients who received systemic corticosteroid as postoperative therapy and compared these values to those who received nasal corticosteroid spray only. RESULTS: No statistically significant difference was noted in the parameters of age, sex, extent of disease, and preoperative serum IgE levels when these values were compared in the group of patients who had recurrence (8 patients) with the group of patients who did not (32 patients). No statistically significant difference was found in the incidence of recurrence in patients in whom systemic corticosteroids were used postoperatively (17 patients) compared with patients who used nasal corticosteroid spray only (15 patients). However, when the patient had a recurrence, when it occurred it was earlier and more severe in patients who used nasal corticosteroid spray only (4 patients). CONCLUSIONS: At the present time, it is not possible to predict recurrence using parameters of age, sex, extent of disease, and serum IgE levels. Larger number of patients preferably in a prospective multicenter meta study are required to address this issue. Though use of systemic corticosteroid does not decrease the incidence of recurrence, it may delay the onset of recurrence and decrease the severity of recurrence. PMID- 15523452 TI - Oral dextromethorphan reduces perioperative analgesic administration in children undergoing tympanomastoid surgery. AB - OBJECTIVE: To determine whether oral dextromethorphan (1 mg/kg) given one hour prior to surgery decreases opioid administration in the perioperative period in children undergoing tympanomastoid surgery. METHODS: This was a prospective randomized double-blinded and placebo-controlled study in which 20 male and 18 female children (age 11.5 +/- 3.5 years) were enrolled. Nineteen children received dextromethorphan (DM), while the other 19 received placebos. Postoperative pain was assessed using a visual analogue scale and a pain score of > or =5 was treated with intravenous morphine sulfate. Patients were discharged home on oral oxycodone. RESULTS: The total doses of fentanyl administered during surgery were higher in the placebo group compared to the DM group (4.1 +/- 2 vs 2.6 +/- 1.4 microg/kg, P = 0.02) and the total doses of intravenous morphine administered in the postoperative period were also higher in the placebo group compared to the DM group (150 +/- 80 vs 73 +/- 56 microg/kg, P = 0.004). The placebo group had a higher pain score at the time of admission to the Day Surgery Unit (DSU) and a higher maximum pain score, compared to the DM group, during their combined stay in the Post-Anesthesia Care Unit and DSU (7.3 +/- 1.5 vs 3.1 +/- 2.6, P = 0.001). CONCLUSIONS: Premedication with DM reduces the need for opioid administration in the perioperative period in children undergoing tympanomastoid surgery. EBM RATING: A. PMID- 15523453 TI - Starplasty: revisiting a pediatric tracheostomy technique. AB - OBJECTIVE: To determine the efficacy the "starplasty" pediatric tracheostomy technique in reducing the incidence of major complications and tracheotomy related death. METHODS: Retrospective chart analysis of all the cases of starplasty performed at 2 tertiary care centers between 1990 and 2002. RESULTS: There were 94 children in our cohort ranging in age from 2 days to 14 years. Of the patients, 47 (50%) were females and 47 (50%) were males and 60 of the children (64%) were younger than 1 year of age. Forty-one patients (44%) had neurologically related airway problems as their primary indication for tracheostomy, 34 (36%) had upper airway obstruction, and the remainder had pulmonary diseases, prolonged intubation, or metabolic-related airway problems. There were 41 short-term complications including 5 cases of tracheal tube dislodgement. There were no instances of pneumothorax or tracheostomy-related death. There were 26 long-term complications. There were no cases of clinically relevant suprastomal collapse that compromised decannulation and no instances of tracheal stenosis. Twenty-six patients underwent decannulation, all of whom developed a tracheocutaneous fistula (TCF). Two patients had spontaneous closure of the TCF; 9 patients underwent surgical repair of their fistulas, 53 patients remain tracheostomy-dependent, and 8 patients died of their primary disease. CONCLUSION: The need for pediatric tracheotomy has increased as a consequence of our success in treating chronically ill children. Starplasty reduces the incidence of major complications, including pneumothorax and death from accidental decannulation. Its major drawback is the need for secondary reconstruction of a tracheocutaneous fistula. EBM RATING: C. PMID- 15523454 TI - Electrocautery versus cold knife technique adenotonsillectomy: a cost analysis. AB - OBJECTIVE: Much has been written about the merits of various techniques of adenotonsillectomy. Proponents of each technique tout many virtues over one another. However, cost remains one variable that has not been thoroughly addressed. In this study, we compared the surgery time, anesthesia time, bleeding, and cost between 2 well-established techniques. Electrocautery tonsillectomy/adenoid ablation was compared against traditional cold knife dissection with adenoid curette. STUDY DESIGN AND SETTING: A retrospective chart review of 275 patients in a large tertiary teaching hospital. RESULTS: The electrocautery group had overall savings of 8, 8, and 9 minutes in surgery, anesthesia, and operating room times, respectively. In terms of cost, the variable cost of the electrocautery group was 19% less than the cold knife dissection group. CONCLUSION/SIGNIFICANCE: This study demonstrated that cautery ablation of adenoids, when employed as part of adenotonsillectomy, enables the surgical team to save significant amount of time, and substantial cost for patients. EBM RATING: B-3. PMID- 15523455 TI - Polysomnography in children scheduled for adenotonsillectomy. AB - OBJECTIVE: Several studies suggest that a reliable diagnosis of childhood sleep disordered breathing (SDB) requires polysomnography (PSG). We compared clinical and PSG-based diagnoses in children scheduled for adenotonsillectomy (AT). Parent responses on a validated Pediatric Sleep Questionnaire were used to determine which symptoms could help identify children with clinical diagnoses of SDB but normal PSG. STUDY DESIGN AND SETTING: Thirty-four children aged 5.0 to 12.9 years and scheduled for AT to treat clinically diagnosed sleep-disordered breathing underwent laboratory-based PSG. Results were scored by 3 different criteria: 1) >1 obstructive apnea (2 breaths or longer) per hour of sleep; 2) >5 apneas or hypopneas per hour of sleep; or 3) >1 apnea, hypopnea, or respiratory event related arousal per hour of sleep. RESULTS: Depending on the criterion used, the PSG documented SDB from a minimum of 18/34 subjects (53%, for criterion I) to as many as 30/34 subjects (88%, for criterion III). Among symptoms studied, absence of daytime mouth breathing and habitual snoring were most helpful in identification of children who had no evidence of SDB on PSG, by criterion I (Chi square, P < 0.05). The absence of other common symptoms, such as "loud snoring" or "trouble breathing" at night, were not helpful. CONCLUSION: Children with clinical diagnoses of SDB may not consistently meet PSG criteria for this disorder. Questions about daytime mouth breathing and habitual snoring might help clinicians recognize children who would not have SDB on objective testing. SIGNIFICANCE: Clinical identification of SDB confirmable on PSG could be improved. However, available outcome data do not yet clarify whether clinical or PSG criteria best identify children likely to suffer morbidity from SDB. EBM RATING: C. PMID- 15523456 TI - Contact diode laser tonsillectomy in children. AB - OBJECTIVE: To compare contact diode laser (CDL) tonsillectomy to monopolar cautery (MPC) with regard to pain and other morbidity during recovery. STUDY DESIGN AND SETTING: Single-blind, randomized trial in a tertiary-care pediatric hospital. Tonsillectomy was performed by CDL in 30 children and by MPC in 28. Parents completed a daily questionnaire for 10 days after surgery. RESULTS: There was significantly less pain in the CDL group than in the MPC group. The CDL group also required much less medication, and was much less likely to experience multiple awakenings (P < 0.001 for all comparisons). CONCLUSION: Recovery after CDL tonsillectomy was associated with much less pain and discomfort than after MPC surgery. SIGNIFICANCE: Postoperative pain is a major concern for surgeons, patients, and their parents. The long recovery period also has costs in school days missed for the child and work missed for the parents. CDL tonsillectomy may greatly mitigate these burdens. PMID- 15523457 TI - Intracranial blood flow velocities and CO2 reactivity in diagnosing migraine related dizziness. AB - OBJECTIVE: Migraine-related dizziness can present with or without headache, often making diagnosis difficult. Flow velocity and CO 2 reactivity testing using transcranial Doppler (TCD) measurement of intracranial blood flow has been described as abnormal in patients with classic or common migraine. This study sought to determine the utility, if any, of this noninvasive technique in the diagnosis of migraine-related dizziness. STUDY DESIGN AND SETTING: A prospective, controlled study in academic neurotology and neurology practices was conducted. Nine patients with migraine-related dizziness and 10 patients with no history of migraine or dizziness were tested with transcranial Doppler ultrasound in a blinded fashion. RESULTS: No statistically significant differences in intracranial blood flow velocities or in cerebrovascular reactivity to hypocapnia were found between patients and controls. CONCLUSIONS: Assessment of intracranial blood flow velocity and CO 2 reactivity using TCD does not help in the diagnosis of migraine-related dizziness. SIGNIFICANCE: A reliable objective test for the diagnosis of migraine-related dizziness remains elusive, and the diagnosis of this patient group continues to be suboptimal. PMID- 15523458 TI - 360-Degree canalith repositioning procedure for the horizontal canal. AB - OBJECTIVES: To evaluate the efficacy of modifications to the standard canalith repositioning procedure (CRP) in the treatment of benign positional vertigo (BPV) of the horizontal semicircular canal (HSC). STUDY DESIGN AND SETTING: Prospective trial of 72 patients with BPV of the HSC treated with a modified 360-degree CRP. RESULTS: 51 of the 62 patients treated for canalolithiasis made a complete recovery (82.2%) after a single 360-degree CRP. CONCLUSIONS AND SIGNIFICANCE: It is proposed a full 360-degree rotation of the HSC involved. It is essential to begin rotation of the patient departing from the affected side towards the healthy side, especially for canal side cupulolithiasis and for canalithiasis of the ampullar branch of the HSC. The head resting on the chest at angle of 30 degree during rotating encourages free-floating debris in the HSC to move into the utricle. Instead of using an oscillator for oscillation of the mastoid bone the head is shaken manually. PMID- 15523460 TI - CO2 laser stapedotomy with the "one-shot" technique--clinical results. AB - OBJECTIVE: To further optimize the surgical technique with the CO 2 laser in stapes surgery a scanner system was used to obtain a footplate perforation of 0.5 to 0.6 mm with only 1 laser application ("one-shot" stapedotomy). STUDY DESIGN: 188 patients with otosclerosis were submitted to a primary CO 2 laser stapedotomy with the SurgiTouch scanner. This study surveys the surgical technique and clinical results. RESULTS: An adequately large perforation diameter could be achieved with a single shot in 68% of the patients treated with the SurgiTouch scanner. In 11% of the patients, a second laser application at the same site was necessary. In 21% of the patients, the perforation had to be enlarged by several slightly overlapping laser applications without scanner. The clinical data of this study clearly documents that there is no evidence of laser depending inner ear affections. CONCLUSION: The CO 2 laser combined with modern scanner systems is well suited for application in stapes surgery. PMID- 15523459 TI - Preliminary functional results of tympanoplasty with titanium prostheses. AB - OBJECTIVE: To assess the use of titanium prostheses in tympanoplasty for reconstruction of the sound pressure and to evaluate the audiometric results obtained. STUDY DESIGN AND SETTING: and patients Retrospective chart review in a tertiary referral center. Between September 1998 and October 2000, 23 titanium middle ear implants (23 TORP) were used for reconstruction of the ossicular chain after tympanomastoidectomy. Preliminary functional results were obtained in 23 cases. RESULTS: Average follow-up time was 18 months (range, 12-40) and no adverse reactions or extrusions occurred. Seven of 16 canal wall down tympanoplasties (43.7%), and 3 of 7 canal wall up tympanoplasties performed (42.9%) had a postoperative air-bone gap of between 0 and 10 dB. CONCLUSIONS: Preliminary results show that titanium prosthesis provide a promising hearing result after total ossicular reconstruction, although long-term results are not yet available. The low weight of these prostheses may be the reason for the decrease in the air-bone gap. PMID- 15523461 TI - Recurrent pneumoparotid: cause and treatment. AB - OBJECTIVE: To document the etiology and successful treatment of severe recurrent pneumoparotid. STUDY DESIGN: Computed tomography, ductal measurement by probe size, surgical treatment. RESULTS: We performed a superficial parotidectomy for a 13-year-old with a history of recurrent parotitis. He subsequently developed recurrent pneumoparotid in the contralateral gland with subcutaneous dissection of air into the face, neck, and mediastinum. At surgery, Stensen's ducts were measured and found to be abnormally patent bilaterally compared to standardized norms. Parotid duct ligation, commonly used for sialorrhea, was employed as a novel treatment and was curative. CONCLUSION: Insufflation of air into the parotid duct system can trouble woodwind instrument players, can complicate dental procedures, or can be self-induced. It is generally a benign condition requiring no therapy. Occasionally, pneumoparotid can be recurrent and lead to inflammation and infection of the parotid or subcutaneous emphysema. SIGNIFICANCE: In selected cases of recurrent pneumoparotid, ductal ligation may be curative. PMID- 15523462 TI - Nasopharyngeal tuberculosis: manifestations between 1991 and 2000. AB - OBJECTIVES/HYPOTHESIS: To present the clinical manifestation of nasopharyngeal tuberculosis. STUDY DESIGN: Clinical analysis of 23 patients with pathologically confirmed nasopharyngeal tuberculosis was carried out retrospectively. SETTING: Srinagarind Hospital, Khon Kaen University. Thailand. RESULTS: The most common presenting symptom was cervical lymphadenopathy (91.3%). The common locations of nodes were the superior and middle cervical. The abnormalities of the nasopharynx were found in 16 patients. The pathological findings were caseous granuloma with positive acid-fast bacilli (AFB) in 15 cases, caseous granuloma with negative AFB in 3 cases, and chronic granulomatous inflammation with negative AFB in 5 cases. Pulmonary tuberculosis was found in 8 of 18 patients. Sixteen patients who received complete treatment responded well. CONCLUSION: Nasopharyngeal tuberculosis commonly presents with cervical lymphadenopathy. The differential diagnosis of tuberculosis from nasopharyngeal carcinoma is difficult. In the patients who have cervical lymphadenopathy and no other identified causes, biopsy of nasopharynx would give an additive information for diagnosis. PMID- 15523463 TI - Otorhinolaryngology or otolaryngology? An etymological approach. AB - The variety and the confusion over the name of many medical terms, including otorhinolaryngological ones prompted us to search their etymology. The term "otorhinolaryngology" (combined form from the ancient Greek words :ous, rhina, larynx, and logos) and the term indicating the specialized doctor who practices "0torhinolaryngology," widely known as ENT in a brief version, are being examined. The investigation leads us to see which of these terms is the appropriate and etymologically correct one, and to support clearly the term "otorhinolaryngology" instead of "otolaryngology" and "otorhinolaryngologist" instead of "ENT." PMID- 15523464 TI - Persistent cough: prevalence of gastroesophageal reflux and study of relevant laryngeal signs. AB - Cough is one of the atypical manifestations of gastroesophageal reflux disorder (GERD). The aim of this study was to evaluate the prevalence of GERD among patients presenting with persistent cough. The study included 80 patients over a period of 3 years. The inclusion criteria were nonsmoker adults with normal chest radiograph whose chief complaint was cough for at least 4 weeks duration. All patients included were subjected to nasal endoscopy, laryngoscopy, and 24-hour pH monitoring. Reflux was recorded in the different positions. Laryngeal signs of reflux were traced for and their significance was calculated. Patients who proved to have GERD received antireflux treatment with a follow-up of 3 months. The response to antireflux treatment was assessed according to subjective and objective improvement. Relapse in the follow-up period occurred in 9% of patients. It is concluded that laryngeal signs of GERD should be well known to diagnose and properly manage persistent cough. PMID- 15523465 TI - Surface electromyographic studies of swallowing in normal subjects: a review of 440 adults. Report 2. Quantitative data: amplitude measures. AB - OBJECTIVES: Surface electromyographic (EMG) studies were performed on 420 normal adults to establish normative database for muscle activity's amplitude during swallowing and drinking clinically useful for ENT department. STUDY DESIGN: Prospective observational study of healthy volunteers. METHODS: Parameters evaluated during swallowing include the EMG activity of the orbicularis oris, masseter, submental, and infrahyoid muscle groups covered by platysma (range, mean, SD). Four tests were examined including voluntary swallow of saliva ("dry" swallow), voluntary single water swallows as normal (variable volume of a bolus), voluntary single swallows of fixed amount of water (20 mL), and continuous drinking of 100 mL of water. Activity of the above-mentioned muscles during swallowing was measured for groups of adults of different age groups. Groups included volunteers and were separated into age groups as follows: ages 18-30, 31 40, 41-50, 51-60, 61-70, and 70+ years of age. RESULTS: Normative data for electric muscle activity (mean +/- SD; range, in microV) during single swallowing and continuous drinking are established for healthy adults. The range of submental group activity during swallows showed a significant decrease with the age, whereas the range of masseter activity changed insignificantly (1 dimensional analysis of variance, SPSS, Chi-square criterion, 95% confidence interval). There were no significant changes in mean muscle activity between different age groups. These parameters represent activities required for normal deglutition, and can be used to identify abnormalities in ENT patients, and provide a basis for comparison of swallowing performance both within and between patients. These measurements can be performed in a simple way, cause no discomfort, do not use radiation, and are noninvasive. The relevance and clinical utility of new and alternative measures, in particular, are discussed. CONCLUSIONS: In single swallow tests range of electric activity of involved muscles is more informative than its mean. In the continuous drinking test, the mean electric activity is the only electric variance that can be evaluated. Orbicularis oris muscle activity is not important when reflex phase of a swallow is evaluated. Surface EMG of swallowing is a simple and reliable method for evaluation of swallowing with a low level of discomfort during the examination. The method can be easily used in outpatient ENT departments for quick evaluation of patients. The normative amplitude data can be used for comparison purposes in preoperaive and postoperative stages and in EMG monitoring during ENT treatment. PMID- 15523466 TI - A new immune-competent animal model of mucosally derived squamous cell carcinoma. AB - OBJECTIVES: To develop an immune-competent animal model for mucosally derived squamous cell carcinoma (SCCA). STUDY DESIGN: Fifteen Fischer 344 rats were inoculated with 1, 2, 5, 10, or 20 x 10(6) FAT7 cells in their flanks. The animals were observed for tumor growth and metastasis. RESULTS: All animals developed tumors that grew exponentially. Pulmonary metastases developed in all animals and 13% developed lymph node metastases. CONCLUSION: The FAT7 flank tumor in Fischer 344 rats is a new animal model that closely resembles the behavior of human mucosal head and neck cancer. SIGNIFICANCE: The existence of an immune competent, mucosally derived, and reliable animal model of SCCA that somewhat resembles human head and neck SCCA gives the opportunity to perform immune modulating experiments on head and neck cancer in these animals. EBM RATING: B-3. PMID- 15523467 TI - Lymphatic malformation of the sphenoid sinus in a pediatric patient. AB - Lymphatic malformation is a benign vascular lesion resulting from lymphatic tissue being isolated from the remainder of the lymphatic system. They are present at birth and up to 90% are diagnosed by 2 years of age. More aggressive lesions are usually diagnosed earlier, with low-grade lesions presenting later with fewer complications. These lesions are hamartomas and not true neoplasms. The term hamartoma is used to describe an abnormally large mass of histologically normal tissue in a normal location. The lymphatic malformation is composed of lymph-filled channels lined with a single layer of flat endothelial cells on a basement membrane. They present with either generalized edema and poorly defined borders (microcystic) or a localized area of multilocular cysts (macrocystic). The term lymphatic malformation has replaced many other outdated terms, such as lymphangioma, cystic hygroma, lymphangioma circumscriptum, and lymphangiomatosis. In this study, we present a case report of a pediatric lymphatic malformation of the sphenoid sinus. To our knowledge, this lesion has not been described in the pediatric population and has been described only once in an adult in the German literature. EBM RATING: C. PMID- 15523468 TI - Chondrosarcoma of the hyoid bone. PMID- 15523469 TI - Osteosarcoma of the larynx. PMID- 15523470 TI - Rhabdomyosarcoma presenting as an acute suppurative mastoiditis. PMID- 15523471 TI - Pleomorphic adenoma of the hard palate. PMID- 15523472 TI - Celebrating the death of proteins. PMID- 15523473 TI - The how and Y of cold shock. PMID- 15523474 TI - Another detour on the Toll road to the interferon antiviral response. PMID- 15523475 TI - Another piece in the transcription initiation puzzle. PMID- 15523476 TI - Opening the GAP. PMID- 15523477 TI - Take two. PMID- 15523479 TI - Regulated nucleosome mobility and the histone code. AB - Post-translational modifications of the histone tails are correlated with distinct chromatin states that regulate access to DNA. Recent proteomic analyses have revealed several new modifications in the globular nucleosome core, many of which lie at the histone-DNA interface. We interpret these modifications in light of previously published data and propose a new and testable model for how cells implement the histone code by modulating nucleosome dynamics. PMID- 15523480 TI - Assembling the mitochondrial outer membrane. AB - The general preprotein translocase of the outer mitochondrial membrane (TOM complex) transports virtually all mitochondrial precursor proteins, but cannot assemble outer-membrane precursors into functional complexes. A recently discovered sorting and assembly machinery (SAM complex) is essential for integration and assembly of outer-membrane proteins, revealing unexpected connections to mitochondrial evolution and morphology. PMID- 15523481 TI - SRP meets the ribosome. AB - Cotranslational targeting directly couples synthesis of proteins to their translocation across or insertion into membranes. The signal recognition particle (SRP) and its membrane-bound receptor facilitate the targeting of the translation machinery, the ribosome, via recognition of a signal sequence in the nascent peptide chain. By combining structures of free and ribosome-bound SRP we derive a structural model describing the dynamic nature of SRP when it meets the ribosome. PMID- 15523483 TI - Copper absorption from foods labelled intrinsically and extrinsically with Cu-65 stable isotope. AB - OBJECTIVE: To determine copper absorption from copper containing foods labelled either intrinsically or extrinsically with a highly enriched Cu-65 stable isotope label. DESIGN: A longitudinal cross-over study. SETTING: The study was conducted at the Institute of Food Research, Human Nutrition Unit, Norwich, UK. SUBJECTS: Subjects were recruited locally via advertisements placed around the Norwich Research Park. A total of 10 volunteers (nine female, one male) took part in the study, but not all volunteers completed each of the test meals. INTERVENTIONS: A highly enriched Cu-65 stable isotope label was administered to volunteers in the form of a reference dose or in breakfast test meals consisting of red wine, soya beans, mushrooms or sunflower seeds. Faecal monitoring and mass spectrometry techniques were used to estimate the relative quantities of copper absorbed from the different test meals. RESULTS: True copper absorption from the reference dose (54%) was similar to extrinsically labelled red wine (49%) and intrinsically labelled sunflower seeds (52%), but significantly higher than extrinsically labelled mushrooms (35%), intrinsically (29%) and extrinsically (15%) labelled soya beans and extrinsically labelled sunflower seed (32%) test meals. CONCLUSIONS: The use of Cu-65 extrinsic labels in copper absorption studies requires validation according to the food being examined; intrinsic and extrinsic labelling produced significantly different results for sunflower seeds. PMID- 15523482 TI - Urinary isoflavone excretion as a compliance measure in a soy intervention among young girls: a pilot study. AB - OBJECTIVE: To investigate the compliance of young girls with a soy intervention. DESIGN: An 8-week dietary intervention and urine sample collection. SETTING: Free living girls. SUBJECTS: A convenience sample of 8- to 14-y-old girls (20 started and 17 finished the study) recruited through flyers distributed to staff members and previous study participants. INTERVENTION: The girls consumed one daily serving of soymilk, soy nuts, or tofu, completed 3-day food records, kept daily soy intake logs, and collected weekly urine samples. MAIN OUTCOME MEASURES: Compliance with the intervention was evaluated by daily soy intake logs, 3-day food records analyzed by the center's Food Composition and Food Groups Servings Databases, and weekly urinary isoflavone excretion using high-pressure liquid chromatography. The statistical analysis included paired t-tests, analysis of variance, and Spearman's rank-order correlation coefficients. RESULTS: Daily soy intake logs indicated a mean intake of 6.28 servings out of a maximum of 7.0 servings per week. The food records revealed a six-fold increase in isoflavone intake during the study period (P<0.01) which was confirmed by an increase in urinary isoflavone excretion of similar magnitude (23.3-142.1 nmol/mg creatinine, P=0.02). CONCLUSIONS: This study demonstrated the ability of young girls to consume one daily soy serving and the usefulness of urinary isoflavones as a primary compliance measure. The high urinary isoflavone excretion levels detected in girls as compared to adult women suggest less intestinal degradation and/or greater absorption of isoflavones in nonadult populations. This finding requires further investigations into the pharmacokinetics of isoflavones. PMID- 15523484 TI - Comparability of dietary patterns assessed by multiple dietary assessment methods: results from the 1946 British Birth Cohort. AB - OBJECTIVE: The aim of this study is to evaluate the consistency of dietary patterns assessed through the use of a dietary recall and a 5-day food diary. DESIGN: Participants (n = 2265) of a longitudinal study of health and development completed 48-h dietary recall at interview, followed by a 5-day food diary and with the 24 h immediately preceding the interview analysed separately as a 24-h recall. Mean intakes of foods and nutrients were calculated and dietary patterns were assessed using exploratory factor analysis, using the method of principal components. Paired t-tests and correlation coefficients were used to compare the three dietary assessment methods. RESULTS: Five distinct dietary patterns were identified using the food diary and the 48-h recall but were less consistent on the 24-h recall. Correlations between factor scores on the 48-h recall and the food diary (r = 0.13-0.67) were higher than those between the 24-h recall and food diary (r = -0.01-0.59). The recall methods were effective at ranking subjects according to food and nutrient intakes, with the 48-h recall and food diary showing higher correlations in both males and females. CONCLUSIONS: This study indicates that a 48-h recall effectively characterises dietary patterns in British adults when compared to a food diary and ranks participants appropriately with respect to most nutrients and foods and is superior to a single 24-h recall. These results have implications for longitudinal studies where maximising response rates to repeat dietary assessment tools is essential. PMID- 15523485 TI - Postprandial insulin response and mitochondrial oxidation in obese men nutritionally treated to lose weight. AB - Obesity, hyperglycemia, and insulin resistance have been associated to an oxidative mitochondrial dysfunction. The aim of this research was to evaluate the relation between carbohydrate metabolism and mitochondrial oxidation, as affected by the weight status and the weight loss induced by a calorie-restricted diet. Lean control men (BMI<25 kg/m2, n = 6) and obese men (BMI>30 kg/m2, n = 14), who were characterized as insulin resistant (n = 6) or insulin sensitive (n = 8) based on HOMA index values, participated in the trial. Plasma insulin levels and mitochondrial oxidation estimated by the 2-keto(1-13C)isocaproate breath test, were measured after ingestion of a test meal during 3 h. Obese subjects repeated the breath test protocol after a 10-week caloric restriction diet to lose weight. Postprandial insulin secretion tended to be marginally higher (P = 0.059) in both obese groups than in controls, while the rate of postprandial mitochondrial oxidation was markedly decreased (P = 0.019) in the obese subjects as compared with lean individuals. The nutritionally induced weight loss produced a rise in the postprandial oxidative process in volunteers initially considered as insulin resistant (P = 0.036), while no statistical differences in the insulin-sensitive obese (P = 0.241) were found. Interestingly, the percentage of oxidized tracer was inversely related to postprandial insulin secretion (r = -0.56; P = 0.001). In conclusion, these results support the hypothetized relation between carbohydrate metabolism and mitochondrial oxidation at a postprandial state in obese subjects, raising interest about mitochondria stimulation as a target in the therapy of obesity. PMID- 15523486 TI - Veganism and its relationship with insulin resistance and intramyocellular lipid. AB - OBJECTIVE: To test the hypothesis that dietary factors in the vegan diet lead to improved insulin sensitivity and lower intramyocellular lipid (IMCL) storage. DESIGN: Case-control study. SETTING: Imperial College School of Medicine, Hammersmith Hospital Campus, London, UK. SUBJECTS: A total of 24 vegans and 25 omnivores participated in this study; three vegan subjects could not be matched therefore the matched results are shown for 21 vegans and 25 omnivores. The subjects were matched for gender, age and body mass index (BMI). INTERVENTIONS: Full anthropometry, 7-day dietary assessment and physical activity levels were obtained. Insulin sensitivity (%S) and beta-cell function (%B) were determined using the homeostatic model assessment (HOMA). IMCL levels were determined using in vivo proton magnetic resonance spectroscopy; total body fat content was assessed by bioelectrical impedance. RESULTS: There was no difference between the groups in sex, age, BMI, waist measurement, percentage body fat, activity levels and energy intake. Vegans had a significantly lower systolic blood pressure ( 11.0 mmHg, CI -20.6 to -1.3, P=0.027) and higher dietary intake of carbohydrate (10.7%, CI 6.8-14.5, P<0.001), nonstarch polysaccharides (20.7 g, CI 15.8-25.6, P<0.001) and polyunsaturated fat (2.8%, CI 1.0-4.6, P=0.003), with a significantly lower glycaemic index (-3.7, CI -6.7 to -0.7, P=0.01). Also, vegans had lower fasting plasma triacylglycerol (-0.7 mmol/l, CI -0.9 to -0.4, P<0.001) and glucose (-0.4 mmol/l, CI -0.7 to -0.09, P=0.05) concentrations. There was no significant difference in HOMA %S but there was with HOMA %B (32.1%, CI 10.3 53.9, P=0.005), while IMCL levels were significantly lower in the soleus muscle ( 9.7, CI -16.2 to -3.3, P=0.01). CONCLUSION: Vegans have a food intake and a biochemical profile that will be expected to be cardioprotective, with lower IMCL accumulation and beta-cell protective. PMID- 15523487 TI - The nutritional status of 1081 elderly people residing in publicly funded shelter homes in Peninsular Malaysia. AB - OBJECTIVES: The aim of this study was to determine the: (1) prevalence of undernutrition as determined by the 'DETERMINE Your Nutritional Health Checklist' (NHC) and (2) factors independently associated with undernutrition among the older residents of these publicly funded shelter homes in Peninsular Malaysia. DESIGN: A total of 1081 elderly people (59%M) over the age of 60 y were surveyed using questionnaires determining baseline demographics, nutritional and cognitive status, physical function and psychological well-being. SETTING: Shelter homes, Peninsular Malaysia. RESULTS: In all, 41.4% (n = 447) were nourished (score <3), 32.1% (n = 347) at moderate risk (score between 3 and 5) and 26.6% (n = 287) were at high risk of undernutrition (score>5) according to the NHC. A large proportion of subjects were underweight with 14.3% of subjects recording a low body mass index (BMI) <18.5 kg/m2 and a further 18.2% recording a BMI between 18.5 and 20 kg/m2. The residential geriatric depression score (GDS-12R) (relative risk (RR) = 1.03 (95% confidence interval (CI) 1.01-1.05); P = 0.002) and the number of illnesses (RR = 1.14 (95% CI 1.07-1.21); P < 0.001) were found to be independently associated with nutritional risk (NHC score > or = 3). Using a BMI < 18.5 kg/m2 as an objective marker for nutritional risk, the NHC was shown to have a sensitivity of 66.4% (95% CI 58.0-74.2%), specificity of 42.7% (95% CI 39.3-46.1%), positive predictive value of 16.2% (95% CI 13.3-19.5%) and a negative predictive value of 88.4% (95% CI 84.9-91.4%). CONCLUSIONS: Many elderly people residing in publicly funded shelter homes in Malaysia may be at-risk of undernutrition, and were underweight. The NHC is better used as an awareness tool rather than as a screening tool. PMID- 15523488 TI - Urinary isoflavonoid excretion is inversely associated with the ratio of protein to dietary fibre intake in young women. AB - OBJECTIVE: To examine the prevalence of excretion of urinary isoflavonoids in women and determine any relationships with accustomed macronutrient intake. DESIGN: Volunteers in one of two 4-month studies. Study 1 was a randomised crossover study whereby subjects consumed a placebo or isoflavone supplement for 2 months and crossed over. Study 2 was a parallel design in which subjects consumed a placebo for 1 month and an isoflavone supplement for 3 months. SETTING: All subjects were free-living, healthy volunteers. SUBJECTS: A total of 25 (study 1, n=14; study 2, n=11) premenopausal women were recruited through advertisements. INTERVENTIONS: Volunteers were supplemented for 2 months (study 1) or 3 months (study 2) with purified isoflavones (86 mg/day) derived from red clover. Urinary isoflavonoids were measured during the placebo and the second month of isoflavone treatment. Macronutrient intakes were determined from weighed food records. RESULTS: During isoflavone supplementation, the concentration of urinary total isoflavonoids increased by 15-fold (P<0.0001), with 5.4-fold variation between individuals. Multiple linear regression analysis showed that 24% of this variation could be explained by an interaction between dietary fibre and protein (P=0.047), with a highly significant inverse association between total isoflavonoid concentration and the protein to fibre ratio (r=-0.51, P=0.009). CONCLUSIONS: Supplementation with purified isoflavones results in an increase in urinary isoflavonoid excretion and part of the individual variation in response is associated with an interaction between intakes of protein and dietary fibre. Whether manipulation of these macronutrients could enhance efficacy of isoflavone supplements remains to be determined. PMID- 15523489 TI - Female migration rate might not be greater than male rate. PMID- 15523490 TI - Epistatic interaction between the monoamine oxidase A and serotonin transporter genes in anorexia nervosa. AB - The serotonin (5-HT) and norepinephrine (NE) systems are likely involved in the aetiology of anorexia nervosa (AN) as sufferers are premorbidly anxious. Specifically, we hypothesize that genes encoding proteins, which clear 5-HT and NE from the synapse, are prime candidates for affecting susceptibility to AN. Supporting our hypothesis, we earlier showed that the NE transporter (NET) and monoamine oxidase A (MAOA) genes appear to contribute additively to increased risk of developing restricting AN (AN-R). With regard to the MAOA gene, a sequence variant that increases MAOA activity and has suggested association with the anxiety condition, panic disorder was preferentially transmitted from parents to affected children. Here we provide evidence in support of interaction between the MAOA and serotonin transporter (SERT) genes in 114 AN nuclear families (patient with AN plus biological parents). A SERT gene genotype with no apparent individual effect on risk and known to be associated with anxiety is preferentially transmitted to children with AN (chi2 trend=9.457, 1 df, P=0.0021) and AN-R alone (chi2 trend=7.477, 1 df, P=0.0063) when the 'more active' MAOA gene variant is also transmitted. The increased risk of developing the disorder is up to eight times greater than the risk imposed by the MAOA gene variant alone -an example of synergistic epistatic interaction. If independently replicated, our findings to date suggest that we may have identified three genes affecting susceptibility to AN, particularly AN-R: the MAOA, SERT, and NET genes. PMID- 15523491 TI - No germline FH mutations in familial breast cancer patients. AB - Fumarate hydratase: (FH) was recently identified as the predisposing gene for a tumor predisposition syndrome, hereditary leiomyomatosis and renal cell cancer (HLRCC) (MIM 605839). In HLRCC, individuals with a germline heterozygous mutation in the FH gene typically develop benign leiomyomas of the skin and the uterus (fibroids, myomas). In a subset of the families, predisposition to renal cell carcinoma and uterine leiomyosarcoma occurs. Other malignancies including breast cancer have also been detected in patients with a germline FH mutation. To examine whether FH could be involved in predisposition to breast cancer, we analyzed germline FH mutations from 85 Finnish breast cancer patients. Most of the cases were selected based on positive family or personal history for malignancies associated with HLRCC. No mutations were found. These results show that FH is not a major predisposing gene for familial breast cancer. PMID- 15523492 TI - A further mutation of the FGFR2 tyrosine kinase domain in mild Crouzon syndrome. AB - We report a family heterozygous for a newly identified mutation in the tyrosine kinase I domain of the FGFR2 gene (1576A > G, encoding the missense substitution Lys526Glu), associated with variable expressivity of Crouzon syndrome, including clinical nonpenetrance. Our observations expand both the clinical and molecular spectrum of this unusual subset of FGFR2 mutations. PMID- 15523493 TI - Uroplakin III is not a major candidate gene for primary vesicoureteral reflux. AB - Vesicoureteral reflux (VUR) is the retrograde flow of urine from the bladder into the ureter and towards the kidneys. VUR is the most common cause of end stage renal failure in both children and adults and it is a major cause of severe hypertension in children. VUR is seen in approximately 1-2% of newborn Caucasians. Substantial evidence exists that VUR is a genetic disorder. Uroplakins are integral membrane proteins found in the bladder wall. Knockout studies in mice have suggested uroplakin III (UPK3) as a candidate gene for VUR. We have used parametric and nonparametric linkage analysis and tests for association, to investigate this possibility in a cohort of 126 sibling pairs affected with primary VUR. None of the analyses showed any substantial evidence for linkage or association of markers at the UPK3 locus to VUR. Our results do not support a role for UPK3 in primary VUR. PMID- 15523494 TI - Phenylbutyrate increases SMN gene expression in spinal muscular atrophy patients. AB - Spinal muscular atrophy (SMA) is caused by insufficient levels of survival motor neuron (SMN) protein. Recently, we found that sodium 4-phenylbutyrate (PB), a well-tolerated FDA approved drug, enhances SMN gene expression in vitro. We provide here the first evidence that oral administration of PB (triButyrate significantly increases SMN expression in leukocytes of SMA patients. This finding provides a strong rationale to further investigate the effects of PB as also supported by preliminary clinical data. PMID- 15523495 TI - Y chromosome evidence for a founder effect in Ashkenazi Jews. AB - Recent genetic studies, based on Y chromosome polymorphic markers, showed that Ashkenazi Jews are more closely related to other Jewish and Middle Eastern groups than to their host populations in Europe. However, Ashkenazim have an elevated frequency of R-M17, the dominant Y chromosome haplogroup in Eastern Europeans, suggesting possible gene flow. In the present study of 495 Y chromosomes of Ashkenazim, 57 (11.5%) were found to belong to R-M17. Detailed analyses of haplotype structure, diversity and geographic distribution suggest a founder effect for this haplogroup, introduced at an early stage into the evolving Ashkenazi community in Europe. R-M17 chromosomes in Ashkenazim may represent vestiges of the mysterious Khazars. PMID- 15523496 TI - PARK11 is not linked with Parkinson's disease in European families. AB - Parkinson's disease (PD) is a genetically heterogeneous disease. Recently, significant linkage has been reported to a 39.5 cM region on the long arm of chromosome 2 (2q36-37; PARK11) in North American Parkinson families under an autosomal dominant model of inheritance. We have performed a replication study to confirm linkage to this region in a European population. Linkage analysis in 153 individuals from 45 European families with a strong family history of PD did not show any significant LOD score in this region. Therefore, PARK11 does not seem to play a major role for familial PD in the European population. PMID- 15523498 TI - Mutations in the known genes are not the major cause of MED; distinctive phenotypic entities among patients with no identified mutations. AB - Multiple epiphyseal dysplasia (MED) is a clinically and genetically heterogeneous chondrodysplasia. Mutations in six genes (COMP, COL9A1, COL9A2, COL9A3, MATN3 and DTDST) have been reported, but the genotype-phenotype correlations and the proportions of cases due to mutations in these genes are still not well characterized. We performed a clinical, radiological and molecular analysis of known MED genes on 29 consecutive MED patients. The mutation analysis resulted in identification of the DTDST mutation in four patients (14%), the COMP mutation in three (10%) and the MATN3 mutation in three (10%). Thus, a disease-causing mutation was identified in 10 patients altogether (34%). The phenotypic features observed in the patients with mutations were in accordance with previously described phenotypes, but two new distinct phenotypic entities were identified in patients in whom no mutation was found. One of them was characterized by severe, early-onset dysplasia of the proximal femurs with almost complete absence of the secondary ossification centres and abnormal development of the femoral necks. The other phenotype was characterized by 'mini-epiphyses', resulting in severe dysplasia of the proximal femoral heads. The findings suggest that mutations in the known genes are not the major cause of MED and are responsible for less than half of the cases. The existence of additional MED loci is supported by the exclusion of known loci by mutation analysis and finding of specific subgroups among these patients. PMID- 15523497 TI - Mutation screening and association analysis of six candidate genes for autism on chromosome 7q. AB - Genetic studies have provided evidence for an autism susceptibility locus (AUTS1) on chromosome 7q. Screening for mutations in six genes mapping to 7q, CUTL1, SRPK2, SYPL, LAMB1, NRCAM and PTPRZ1 in 48 unrelated individuals with autism led to the identification of several new coding variants in the genes CUTL1, LAMB1 and PTPRZ1. Analysis of genetic variants provided evidence for association with autism for one of the new missense changes identified in LAMB1; this effect was stronger in a subgroup of affected male sibling pair families, implying a possible specific sex-related effect for this variant. Association was also detected for several polymorphisms in the promoter and untranslated region of NRCAM, suggesting that alterations in expression of this gene may be linked to autism susceptibility. PMID- 15523499 TI - Haplotype structure of the beta adrenergic receptor genes in US Caucasians and African Americans. AB - The beta-adrenergic receptors (beta-AR) are G protein-coupled receptors activated by epinephrine and norepinephrine and are involved in a variety of their physiological functions. Previously, three beta-AR genes (ADRB1, ADRB2 and ADRB3) were resequenced, identifying polymorphisms that were used in genetic association studies of cardiovascular and metabolic disorders. These studies have produced intriguing but inconsistent results, potentially because the known functional variants: ADRB1 Arg389Gly and Gly49Ser, ADRB2 Arg16Gly and Gln27Glu, and ADRB3 Arg64Trp provided an incomplete picture of the total functional diversity at these genes. Therefore, we created marker panels for each beta-AR gene that included the known functional markers and also other markers evenly spaced and with sufficient density to identify haplotype block structure and to maximize haplotype diversity. A total of 27 markers were genotyped in 96 US Caucasians and 96 African Americans. In both populations and for each gene, a single block with little evidence of historical recombination was observed. For each gene, haplotype captured most of the information content of each functional locus, even if that locus was not genotyped, and presumably haplotype would capture the signal from unknown functional loci whose alleles are of moderate abundance. This study demonstrates the utility of using beta-AR gene haplotype maps and marker panels as tools for linkage studies on beta-AR function. PMID- 15523500 TI - Biological variations, genetic polymorphisms and familial resemblance of TNF alpha and IL-6 concentrations: STANISLAS cohort. AB - Cytokines are involved in the development of several inflammatory diseases and atherosclerosis. Their variations in healthy individuals are not well defined. The aims of this study were: firstly, to identify factors affecting biological variation of interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha); secondly, to study their family resemblance; and thirdly, to evaluate the effect of two TNF-alpha (-308G/A and -238G/A) and two IL-6 polymorphisms (174G/C and 572G/C) on their corresponding circulating levels. A total of 171 healthy families selected from the STANISLAS cohort were studied. Age was negatively related to TNF-alpha concentrations in offspring only (both sons and daughters). Additionally, IL-6 and TNF-alpha levels were differently influenced by gender, white blood cells, tobacco consumption, and HDL-cholesterol level. A weak significant familial resemblance for TNF-alpha concentration was observed in siblings only. There was no significant familial resemblance for IL-6 levels. The TNF-alpha -308A allele was associated with decreased TNF-alpha concentrations in both offspring aged less than 18 and males without overweight (BMI<25 kg/m(2)). Fathers carrying the IL-6 -174CC genotype had higher IL-6 levels than those with the IL-6 -174G allele. Parents with the IL-6 -572GG genotype had higher IL-6 concentrations than the C allele carriers. In this sample of healthy families, plasma levels of IL-6 and TNF-alpha were differently affected by biological parameters including age, gender and smoking, and the impact of their respective polymorphisms was influenced by gender, age and BMI. PMID- 15523501 TI - Diagnostic investigations in individuals with mental retardation: a systematic literature review of their usefulness. AB - There are no guidelines available for diagnostic studies in patients with mental retardation (MR) established in an evidence-based manner. Here we report such study, based on information from original studies on the results with respect to detected significant anomalies (yield) of six major diagnostic investigations, and evaluate whether the yield differs depending on setting, MR severity, and gender. Results for cytogenetic studies showed the mean yield of chromosome aberrations in classical cytogenetics to be 9.5% (variation: 5.4% in school populations to 13.3% in institute populations; 4.1% in borderline-mild MR to 13.3% in moderate-profound MR; more frequent structural anomalies in females). The median yield of subtelomeric studies was 4.4% (also showing female predominance). For fragile X screening, yields were 5.4% (cytogenetic studies) and 2.0% (molecular studies) (higher yield in moderate-profound MR; checklist use useful). In metabolic investigations, the mean yield of all studies was 1.0% (results depending on neonatal screening programmes; in individual populations higher yield for specific metabolic disorders). Studies on neurological examination all showed a high yield (mean 42.9%; irrespective of setting, degree of MR, and gender). The yield of neuroimaging studies for abnormalities was 30.0% (higher yield if performed on an indicated basis) and the yield for finding a diagnosis based on neuroradiological studies only was 1.3% (no data available on value of negative findings). A very high yield was found for dysmorphologic examination (variation 39-81%). The data from this review allow conclusions for most types of diagnostic investigations in MR patients. Recommendations for further studies are provided. PMID- 15523503 TI - Genetics of female functional virginity in the parthenogenesis-Wolbachia infected parasitoid wasp Telenomus nawai (Hymenoptera: Scelionidae). AB - A lepidopteran egg parasitoid species Telenomus nawai consists of two distinct populations with different reproductive modes. One is a completely thelytokous population consisting of females only, whereas the other displays arrhenotokous reproduction where fertilized eggs develop into diploid females and unfertilized eggs into haploid males. Thelytoky in T. nawai is caused by a bacterial symbiont, the parthenogenesis-inducing (PI) Wolbachia. Recent theoretical studies have shown that when a PI-Wolbachia is spreading in a population, mutations that allow uninfected females to produce more male offspring will spread rapidly eventually becoming fixed. The consequence of such a mutation is that sexual reproduction is no longer successful in infected females. Here we determine the genetic basis of the females' inability to reproduce sexually by introgressing the genome of a thelytokous line into an arrhenotokous line. The results suggest that the mutations are recessive and inherited either as a single-locus major gene with some modifiers, or as two partially linked loci. PMID- 15523504 TI - Phylogeographic evidence for the existence of an ancient biogeographic barrier: the Isthmus of Kra Seaway. AB - Biogeographic boundaries are characterised by distinct faunal and floral assemblages restricted on either side, but patterns among groups of taxa often vary and may not be discrete. Historical biogeography as a consequence, while providing crucial insights into the relationship between biological diversity and earth history, has some limitations. Patterns of intraspecific molecular variation, however, may show unambiguous evidence for such historical divides, and can be used to test competing biogeographic hypotheses (often based on the dispersal-vicariance debate). Here, we utilise this method to test the hypothesis that a major biogeographic transition zone between the Sundaic and Indochinese biotas, located just north of the Isthmus of Kra in SE Asia, is the result of Neogene marine transgressions that breached the Isthmus in two locations for prolonged periods of time (>1 million year duration). Phylogeographic analyses of a freshwater decapod crustacean, the giant freshwater prawn Macrobrachium rosenbergii, strongly supports the historical existence of the more northerly postulated seaway. Results presented here highlight the power of utilising intraspecific molecular variation in testing biogeographical hypotheses. PMID- 15523505 TI - Ontogenetic change in relative performance of allozyme genotypes influences detection of heterosis in the earthworm Eisenia andrei. AB - The effect of ontogeny on relationships between allozyme genotypes and fresh weight was measured weekly throughout the life history of the earthworm Eisenia andrei to test the hypothesis that there is an ontogenetic component to variation in such relationships. Two of six allozyme loci showed a significant increase in apparent heterosis with ontogeny, while one locus showed a significant decrease in apparent heterosis. Three loci showed a significant decrease in the performance of common homozygotes with ontogeny. Patterns of relative genotypic performance varied among loci, but the cumulative effect was an increase in apparent allozyme heterosis later in ontogeny coinciding with a series of positive relationships between multilocus heterozygosity and fresh weight. The results could not be used to determine whether these patterns were caused by selection acting on the loci directly or on loci tightly linked to allozyme loci. However, because the same individuals were used throughout this study and thus allele frequencies and heterozygote deficiency were constant, the presence of both ontogenetic effects and differences in such patterns among loci is not compatible with a general inbreeding effect. Examining relative genotypic performance repetitively using the same individuals through ontogeny or in different environments is a very powerful experimental design for testing the effects of inbreeding or other populational factors. PMID- 15523506 TI - The evolution of evolution. PMID- 15523507 TI - Local adaptation in the rock pocket mouse (Chaetodipus intermedius): natural selection and phylogenetic history of populations. AB - Elucidating the causes of population divergence is a central goal of evolutionary biology. Rock pocket mice, Chaeotdipus intermedius, are an ideal system in which to study intraspecific phenotypic divergence because of the extensive color variation observed within this species. Here, we investigate whether phenotypic variation in color is correlated with local environmental conditions or with phylogenetic history. First, we quantified variation in pelage color (n=107 mice) and habitat color (n=51 rocks) using a spectrophotometer, and showed that there was a correlation between pelage color and habitat color across 14 sampled populations (R2=0.43). Analyses of mtDNA sequences from these same individuals revealed strong population structure in this species across its range, where most variation (63%) was partitioned between five geographic regions. Using Mantel tests, we show that there is no correlation between color variation and mtDNA phylogeny, suggesting that pelage coloration has evolved rapidly. At a finer geographical scale, high levels of gene flow between neighboring melanic and light populations suggest the selection acting on color must be quite strong to maintain habitat-specific phenotypic distributions. Finally, we raise the possibility that, in some cases, migration between populations of pocket mice inhabiting different lava flows may be responsible for similar melanic phenotypes in different populations. Together, the results suggest that color variation can evolve very rapidly over small geographic scales and that gene flow can both hinder and promote local adaptation. PMID- 15523508 TI - Testing for clonal propagation. AB - The conceptual basis for testing clonal propagation is reconsidered with the result that two steps need to be distinguished clearly: (1) specification of the characteristics of multilocus genotype frequencies that result from sexual reproduction together with the kinds of deviations from these characteristics that are produced by clonal propagation, and (2) a statistical method for detecting these deviations in random samples. It is pointed out that a meaningful characterization of sexual reproduction reflects the association of genes in (multilocus) genotypes within the bounds set by the underlying gene frequencies. An appropriate measure of relative gene association is developed which is equivalent to a multilocus generalization of the standardized gametic disequilibrium (linkage disequilibrium). Its application to the characterization of sexually produced multilocus genotypes is demonstrated. The resulting hypothesis on the frequency of a sexually produced genotype is tested with the help of the (significance) probability of obtaining at least two copies of the genotype in question in a random sample of a given size. If at least two copies of the genotype are observed in a sample, and if the probability is significant, then the hypothesis of sexual reproduction is rejected in favor of the assumption that all copies of the genotype belong to the same clone. Common testing approaches rest on the hypothesis of completely independent association of genes in genotypes and on the (significance) probability of obtaining at least as many copies of a genotype as observed in a sample. The validity of these approaches is discussed in relation to the above considerations and recommendations are set out for conducting appropriate tests. PMID- 15523509 TI - Comparison of recruitment, retention, and compliance results for three children's exposure monitoring studies. AB - The School Health Initiative: Environment, Learning, Disease (SHIELD) study, the Minnesota Children's Pesticide Exposure Study (MNCPES), and the National Cooperative Inner-City Asthma Study (NCICAS) are three of the most intensive and invasive exposure-monitoring projects ever undertaken in children. An intrinsic facet of each study was the need to recruit children and their families, retain them for the duration of the project, and ensure that they completed monitoring protocols successfully. All of the studies used fiscal incentives to encourage participation, retention, and compliance. Recruitment rates varied from 40% in MNCPES, to 57% in SHIELD, to 64% in NCICAS, while retention rates varied from 85% in SHIELD, to 94% in MNCPES, to 95% in NCICAS. Rates of compliance with exposure sampling procedures were typically >80% for each study. For example, > or =85% of the enrolled children provided all requested urine samples (1 for NCICAS, 2 for SHIELD, 3 for MNCPES), and 82% of the children in SHIELD provided two out of two blood samples (optional in MNCPES and NCICAS). However, compliance rates were substantially lower (34% SHIELD, 40% NCICAS, not applicable to MNCPES) for the more complex and time-consuming protocol used to collect peak flow data. Overall, results demonstrate that it is practical and affordable to conduct demanding exposure-monitoring studies in children, including children from poor minority communities. PMID- 15523510 TI - The research career of Richard B. Stein (1940-). AB - This report reviews the research and mentoring career of Richard B. Stein (1940 ). In 1962, he completed a B.S. degree in physics at the Massachusetts Institute of Technology, USA, and thereafter an M.A. (1964), Ph.D. (1966), and postdoctoral training (1966-68) at the University of Oxford, UK. He subsequently assumed a faculty position at the University of Alberta (Canada), where he is currently an active researcher and mentor. To this point in 2004, Stein has trained and collaborated closely with over 160 scientists, largely neuroscientists and biomedical engineers, from 27 countries. He and his former trainees and collaborators have made important contributions on topics that span the cellular to-behavioral spectrum of movement and rehabilitation-prosthetics neuroscience. His mentors, trainees, and collaborators include scientists whose countries of origin are: Australia, 2; Austria, 1; Belgium, 1; Bulgaria, 1; Canada, 64; China, 6; Denmark, 1; Germany, 1; Great Britain, 16; Hong Kong, 4; India, 5; Iraq, 2; Italy, 2; Japan, 10; Kenya, 1; New Zealand, 4; Pakistan, 1; Palestine, 1; Poland, 1; Romania, 1; South Africa, 1; Sri Lanka, 1; The Netherlands, 1; Turkey, 1; Uruguay, 1; USA, 21; and Yugoslavia, 6. In all instances, Stein's research collaborations and mentoring have advanced the careers of his trainees and junior collaborators, a well-deserved and important compliment to a stellar movement neuroscientist. PMID- 15523511 TI - Mechanotransduction in spider slit sensilla. AB - Mechanoreception is a vital constituent of several sensory modalities and a wide range of internal regulatory processes, but fundamental mechanisms for neural detection of mechanical stimuli have been difficult to characterize because of the morphological properties of most mechanoreceptors and the nature of the stimulus itself. An invertebrate preparation, the VS-3 lyriform slit sense organ of the spider, Cupiennius salei, has proved useful because it possesses large mechanosensory neurons, whose cell bodies are close to the sites of sensory transduction, and accessible to intracellular recording during mechanotransduction. This has made it possible to observe and experiment with all the major stages of mechanosensation. Here, we describe several important findings from this preparation, including the estimated number, conductance and ionic selectivity of the ion channels responsible for mechanotransduction, the major voltage-activated ion channels responsible for action potential encoding and control of the dynamic properties of the neurons, the location of action potential initiation following mechanical stimulation, and the efferent control of mechanoreception. While many details of mechanosensation remain to be discovered, the VS-3 system continues to offer important opportunities to advance our understanding of this crucial physiological process. PMID- 15523512 TI - Synaptic integration in motoneurons with hyper-excitable dendrites. AB - Motoneurons have extensive dendritic trees that receive the numerous inputs required to produce movement. These dendrites are highly active, containing voltage-sensitive channels that generate persistent inward currents (PICs) that can enhance synaptic input 5-fold or more. However, this enhancement is proportional to the level of activity of monoaminergic inputs from the brainstem that release serotonin and noradrenalin. The higher this activity, the larger the dendritic PIC and the higher the firing rate evoked by a given amount of excitatory synaptic input. This brainstem control of motoneuron input-output gain translates directly into control of system gain of a motor pool and its muscle. Because large dendritic PICs are probably necessary for motoneurons to have sufficient gain to generate large forces, it is possible that descending monoaminergic inputs scale in proportion to voluntary force. Inhibition from sensory inputs has a strong suppressive effect on dendritic PICs: the stronger the inhibition, the smaller the PIC. Thus, local inhibitory inputs within the cord may oppose the descending monoaminergic control of PICs. Most motor behaviors evoke a mixture of excitation and inhibition (e.g., the reciprocal inhibition between antagonists). Therefore, normal joint movements may involve constant adjustment of PIC amplitude. PMID- 15523513 TI - Possible contributions of CPG activity to the control of rhythmic human arm movement. AB - There is extensive modulation of cutaneous and H-reflexes during rhythmic leg movement in humans. Mechanisms controlling reflex modulation (e.g., phase- and task-dependent modulation, and reflex reversal) during leg movements have been ascribed to the activity of spinal central pattern generating (CPG) networks and peripheral feedback. Our working hypothesis has been that neural mechanisms (i.e., CPGs) controlling rhythmic movement are conserved between the human lumbar and cervical spinal cord. Thus reflex modulation during rhythmic arm movement should be similar to that for rhythmic leg movement. This hypothesis has been tested by studying the regulation of reflexes in arm muscles during rhythmic arm cycling and treadmill walking. This paper reviews recent studies that have revealed that reflexes in arm muscles show modulation within the movement cycle (e.g., phase-dependency and reflex reversal) and between static and rhythmic motor tasks (e.g., task-dependency). It is concluded that reflexes are modulated similarly during rhythmic movement of the upper and lower limbs, suggesting similar motor control mechanisms. One notable exception to this pattern is a failure of contralateral arm movement to modulate reflex amplitude, which contrasts directly with observations from the leg. Overall, the data support the hypothesis that CPG activity contributes to the neural control of rhythmic arm movement. PMID- 15523514 TI - Cross-bridge mechanisms underlying the history-dependent properties of muscle spindles and stretch reflexes. AB - The effects of prior movement on the force responses of skeletal muscle are compared with the effects of movement history on the changes in firing rate of muscle spindle receptors. Prior release results in the linearization of the mechanical properties of skeletal muscles, which can be provisionally explained by cross-bridge models of muscular contraction. The history-dependence of responses of muscle spindle receptors in unanesthetized decerebrate preparations appears to result from the kinetics of cycling and noncycling cross-bridges. The results of this comparison indicate that the integration of mechanical properties of muscle and spindle receptor promotes stiffness regulation. PMID- 15523515 TI - Associations between force and fatigue in fast-twitch motor units of a cat hindlimb muscle. AB - Associations were quantified between the control force and fatigue-induced force decline in 22 single fast-twitch-fatigable motor units of 5 deeply anesthetized adult cats. The units were subjected to intermittent stimulation at 1 train/s for 360 s. Two stimulation patterns were delivered in a pseudo-random manner. The first was a 500-ms train with constant interpulse intervals. The second pattern had the same number of stimuli, mean stimulus rate, and stimulus duration, but the stimulus pulses were rearranged to increase the force produced by the units in the control (prefatigue) state. The associations among the control peak tetanic force of these units, 3 indices of fatigue, and total cumulative force during fatiguing contractions were dependent, in part, on the stimulation pattern used to produce fatigue. The associations were also dependent, albeit to a lesser extent, on the force measure (peak vs. integrated) and the fatigue index used to quantify fatigue. It is proposed that during high-force fatiguing contractions, neural mechanisms are potentially available to delay and reduce the fatigue of fast-twitch-fatigable units for brief, but functionally relevant, periods. In contrast, the fatigue of slow-twitch fatigue-resistant units seems more likely to be controlled largely, if not exclusively, by metabolic processes within their muscle cells. PMID- 15523516 TI - Contribution of sensory feedback to ongoing ankle extensor activity during the stance phase of walking. AB - Numerous investigations over the past 15 years have demonstrated that sensory feedback plays a critical role in establishing the timing and magnitude of muscle activity during walking. Here we review recent studies reporting that sensory feedback makes a substantial contribution to the activation of extensor motoneurons during the stance phase. Quantitative analysis of the effects of loading and unloading ankle extensor muscles during walking on a horizontal surface has shown that sensory feedback can increase the activity of ankle extensor muscles by up to 60%. There is currently some uncertainty about which sensory receptors are responsible for this enhancement of extensor activity, but likely candidates are the secondary spindle endings in the ankle extensors of humans and the Golgi tendon organs in the ankle extensors of humans and cats. Two important issues arise from the finding that sensory feedback from the leg regulates the magnitude of extensor activity. The first is the extent to which differences in the magnitude of activity in extensor muscles during different locomotor tasks can be directly attributed to changes in the magnitude of sensory signals, and the second is whether the enhancement of extensor activity is determined primarily by feedback from a specific group of receptors or from numerous groups of receptors distributed throughout the leg. Limitations of current experimental strategies prevent a straightforward empirical resolution of these issues. A potentially fruitful approach in the immediate future is to develop models of the known and hypothesized neuronal networks controlling motoneuronal activity, and use these simulations to control forward dynamic models of the musculo-skeletal system. These simulations would help understand how sensory signals are modified with a change in locomotor task and, in conjunction with physiological experiments, establish the extent to which these modifications can account for changes in the magnitude of motoneuronal activity. PMID- 15523517 TI - Selective motor unit recruitment via intrafascicular multielectrode stimulation. AB - Recruitment of force via independent asynchronous firing of large numbers of motor units produces the grace and endurance of physiological motion. We have investigated the possibility of reproducing this physiological recruitment strategy by determining the selectivity of access to large numbers of independent motor units through intrafascicular multielectrode stimulation (IFMS) of the peripheral nerve. A Utah Slanted Electrode Array containing 100, 0.5-1.5 mm-long penetrating electrodes was inserted into the sciatic nerve of a cat, and forces generated by the 3 heads of triceps surea in response to electrical stimulation of the nerve were monitored via force transducers attached to their tendons. We found a mean of 17.4 +/- 4.9 (mean +/- SEM) electrodes selectively excited maximal forces in medial gastrocnemius before exciting another muscle. Among electrodes demonstrating selectivity at threshold, a mean of 7.3 +/- 2.7 electrodes were shown to recruit independent populations of motor units innervating medial gastrocnemius (overlap < 20%). Corresponding numbers of electrodes were reported for lateral gastrocnemius and soleus, as well. We used these stimulation data to emulate physiological recruitment strategies, and found that independent motor unit pool recruitment approximates physiological activation more closely than does intensity-based recruitment or frequency-based recruitment. PMID- 15523518 TI - Ankle extensor proprioceptors contribute to the enhancement of the soleus EMG during the stance phase of human walking. AB - A rapid plantar flexion perturbation applied to the ankle during the stance phase of the step cycle during human walking unloads the ankle extensors and produces a marked decline in the soleus EMG. This demonstrates that sensory activity contributes importantly to the enhancement of the ankle extensor muscle activation during human walking. On average, the EMG begins to decline approximately 52 ms after the perturbation. In contrast, a rapid dorsi flex ion perturbation produces a group Ia mediated short-latency stretch reflex burst with an onset latency of approximately 36 ms. The transmission of sensory traffic from the foot and ankle was suppressed in 10 subjects by an anaesthetic nerve block produced with local injections of lidocaine hydrochloride. The anaesthetic block had no effect on the stance phase soleus EMG, the latencies of the EMG responses, or the magnitude of the EMG decline following the plantar flexion perturbation. Therefore, it is more likely that proprioceptive afferents, rather than cutaneous afferents, contribute to the background soleus EMG during the late stance phase of the step cycle. The large difference in onset latencies between the short latency reflex and unload responses suggests that the largest of the active group Ia afferents might not contribute strongly to the background soleus EMG, although it remains to be determined which of the proprioceptive pathways provide the more important contributions. PMID- 15523519 TI - Adaptive changes of locomotion after central and peripheral lesions. AB - This paper reviews findings on the adaptive changes of locomotion in cats after spinal cord or peripheral nerve lesions. From the results obtained after lesions of the ventral/ventrolateral pathways or the dorsal/dorsolateral pathways, we conclude that with extensive but partial spinal lesions, cats can regain voluntary quadrupedal locomotion on a treadmill. Although tract-specific deficits remain after such lesions, intact descending tracts can compensate for the lesioned tracts and access the spinal network to generate voluntary locomotion. Such neuroplasticity of locomotor control mechanisms is also demonstrated after peripheral nerve lesions in cats with intact or lesioned spinal cords. Some models have shown that recovery from such peripheral nerve lesions probably involves changes at the supra spinal and spinal levels. In the case of somesthesic denervation of the hindpaws, we demonstrated that cats with a complete spinal section need some cutaneous inputs to walk with a plantigrade locomotion, and that even in this spinal state, cats can adapt their locomotion to partial cutaneous denervation. Altogether, these results suggest that there is significant plasticity in spinal and supraspinal locomotor controls to justify the beneficial effects of early proactive and sustained locomotor training after central (Rossignol and Barbeau 1995; Barbeau et al. 1998) or peripheral lesions. PMID- 15523520 TI - Anatomical organization of motoneurons and interneurons in the mudpuppy (Necturus maculosus) brachial spinal cord: the neural substrate for central pattern generation. AB - The isolated brachial spinal cord of the mudpuppy is useful for studies of neural networks underlying forelimb locomotion, but information about its anatomy is scarce. We addressed this issue by combining retrograde labeling with fluorescent tracers and confocal microscopy. Remarkably, the central region of gray matter was aneural and contained only a tenuous meshwork of glial fibers and large extracellular spaces. Somata of motoneurons (MNs) and interneurons (INs), labeled retrogradely from ventral roots or axons in the ventro-lateral funiculus, respectively, were confined within a gray neuropil layer abutting the white matter borders, while their dendrites projected widely throughout the white matter. A considerable fraction of labeled INs was found contralaterally with axons crossing beneath a thick layer of ependyma surrounding the central canal. Dorsal roots (DRs) produced dense presynaptic arbors within a restricted dorsal region containing afferent terminations, within which dorsally directed MN and IN dendrites mingled with dense collections of synaptic boutons. Our data suggest that a major fraction of synaptic interactions takes place within the white matter. This study provides a detailed foundation for electrophysiological experiments aimed at elucidating the neural circuits involved in locomotor pattern generation. PMID- 15523521 TI - Activation of NMDA receptors is required for the initiation and maintenance of walking-like activity in the mudpuppy (Necturus Maculatus). AB - We hypothesized that blocking the activation of N-methyl-D-aspartate (NMDA) receptors prevents the initiation of walking-like activity and abolishes the ongoing rhythmic activity in the spinal cord-forelimb preparation from the mudpuppy. Robust walking-like movements of the limb and rhythmic alternating elbow flexor-extensor EMG pattern characteristic of walking were elicited when continuous perfusion of the spinal cord with solution containing D-glutamate. The frequency of the walking-like activity was dose-dependent on the concentration of D-glutamate in the bath over a range of 0.2 to 0.9 mmol/L. Elevation of potassium concentrations failed to induce walking-like activity. Application of the selective antagonist 2-amino-5-phosphonovalerate (AP-5) produced dose-dependent block of the initiation and maintenance of walking-like activity induced by D glutamate. Complete block of the activity was achieved when the concentration of AP-5 reached 20 micromol/L. Furthermore, application of L-701,324 (a selective antagonist of the strychnine-insensitive glycine site of NMDA receptor) (1-10 micromol/L) also resulted in complete block of the walking-like activity. In contrast, application of the non-NMDA receptor antagonist 6-cyno-7 nitroquinoxaline-2,3-dione (CNQX) (1-50 micromol/L) induced a dose-dependent inhibition of the burst frequency but failed to result in a complete block. Only at concentration as high as 100 micromol/L, did CNQX cause complete block of the rhythmic activity, presumably through nonspecific action on the strychnine insensitive glycine site of NMDA receptors. These results suggest that activation of NMDA receptors is required for the initiation and maintenance of walking-like activity. Operation of non-NMDA receptors plays a powerful role in the modulation of the walking-like activity in the mudpuppy. PMID- 15523522 TI - The resilience of the size principle in the organization of motor unit properties in normal and reinnervated adult skeletal muscles. AB - Henneman's size principle relates the input and output properties of motoneurons and their muscle fibers to size and is the basis for size-ordered activation or recruitment of motor units during movement. After nerve injury and surgical repair, the relationship between motoneuron size and the number and size of the muscle fibers that the motoneuron reinnervates is initially lost but returns with time, irrespective of whether the muscles are self- or cross-reinnervated by the regenerated axons. Although the return of the size relationships was initially attributed to the recovery of the cross-sectional area of the reinnervated muscle fibers and their force per fiber, direct enumeration of the innervation ratio and the number of muscle fibers per motoneuron demonstrated that a size-dependent branching of axons accounts for the size relationships in normal muscle, as suggested by Henneman and his colleagues. This same size-dependent branching accounts for the rematching of motoneuron size and muscle unit size in reinnervated muscles. Experiments were carried out to determine whether the daily amount of neuromuscular activation of motor units accounts for the size-dependent organization and reorganization of motor unit properties. The normal size dependent matching of motoneurons and their muscle units with respect to the numbers of muscle fibers per motoneuron was unaltered by synchronous activation of all of the motor units with the same daily activity. Hence, the restored size relationships and rematching of motoneuron and muscle unit properties after nerve injuries and muscle reinnervation sustain the normal gradation of muscle force during movement by size-ordered recruitment of motor units and the process of rate coding of action potentials. Dynamic modulation of size of muscle fibers and their contractile speed and endurance by neuromuscular activity allows for neuromuscular adaptation in the context of the sustained organization of the neuromuscular system according to the size principle. PMID- 15523523 TI - Infant stepping: a window to the behaviour of the human pattern generator for walking. AB - The aim of this paper is to provide evidence, both published and new, to support the notion that human infants are particularly good subjects for the study of the pattern generator for walking. We and others have shown that stepping can be initiated by sensory input from the legs or by general heightened excitability of the infant. New results are presented here to suggest that weight support through the feet and rapid extension of the legs are important proprioceptive inputs to initiate stepping. Our previous work has shown that infants can step at many different speeds when supported on a treadmill. The step cycle duration shortens as the speed increases, with the changes coming largely from the stance phase, just as in most other terrestrial animals. Moreover, we have shown that infants will step in all directions. Regardless of the direction of stepping, the step cycle changes in the same way with walking speed, suggesting the circuitry that controls different directions of walking share common elements. We have also shown that infant stepping is highly organized. Sensory inputs, whether proprioceptive or touch, are gated in a functional way so that only important sensory inputs generate a response. For example, touch to the lateral surface of the foot elicits a response only in sideways walking, and only in the leading limb. New data is presented here to show that the pattern generators from each limb can operate somewhat independently. On a split-belt treadmill with the 2 belts running at different speeds or in different directions, the legs showed considerable independence in behaviour. Yet, the pattern generators on each side interact to ensure that swing phase does not occur at the same time. These studies have provided insight into the organization of the pattern generator for walking in humans. It will be interesting in the future to study how maturation of the descending tracts changes walking behaviour to allow independent bipedal walking. PMID- 15523524 TI - Vestibulospinal influences on lower limb motoneurons. AB - Galvanic vestibular stimulation (GVS) is a research tool used to activate the vestibular system in human subjects. When a low-intensity stimulus (1-4 mA) is delivered percutaneously to the vestibular nerve, a transient electromyographic response is observed a short time later in lower limb muscles. Typically, galvanically evoked responses are present when the test muscle is actively engaged in controlling standing balance. However, there is evidence to suggest that GVS may be able to modulate the activity of lower limb muscles when subjects are not in a free-standing situation. The purpose of this review is to examine 2 studies from our laboratory that examined the effects of GVS on the lower limb motoneuron pool. For instance, a monopolar monaural galvanic stimulus modified the amplitude of the ipsilateral soleus H-reflex. Furthermore, bipolar binaural GVS significantly altered the onset of activation and the initial firing frequency of gastrocnemius motor units. The following paper examines the effects of GVS on muscles that are not being used to maintain balance. We propose that GVS is modulating motor output by influencing the activity of presynaptic inhibitory mechanisms that act on the motoneuron pool. PMID- 15523525 TI - What visual information is used for navigation around obstacles in a cluttered environment? AB - The goal of this study was to determine what visual information is used to navigate around barriers in a cluttered terrain. Twelve traffic pylons were arranged randomly in a 4.55 x 3.15 m travel area: there were 20 different arrangements. For each arrangement, individuals (N = 6) were positioned in 1 of 3 locations on the outside border with their eyes closed: on verbal command they were instructed to open their eyes and quickly go to 1 of 2 specified goals (2 vertical posts defining a door) located on one edge of the travel area. The movement of the body was tracked using the OPTOTRAK system, with the IREDS placed on a collar worn by the subjects. Experimental data of travel path chosen were compared with those predicted by models that incorporated different types of visual information to control path trajectory. The 6 models basically use 2 different strategies for route selection: reactive control based on visual input about the obstacle encountered in the line-of-sight travel path (Model # 1) and path planning based on different visual information (Model # 2, 3, 4, 5, and 6). The models that involve path planning are grouped into 2 categories: models 2, 3, 4, and 5 need detailed geometrical configuration of the obstacles to plan a route while model 6 plans a route based on identifying and avoiding a cluster of obstacles in the travel path. Two measures were used to compare model performance with the actual travel path: the difference in area between predicted and actual travel path and the number of trials that accurately predicted the number of turns during travel. The results suggest that route selection is not based on reactive control, but does involve path planning. The model that best predicts the travel paths taken by the individuals uses visual information about cluster of obstacles and identification of safe corridors to plan a route. PMID- 15523526 TI - The accuracy of perception of a pinch grip force in older adults. AB - The fact that humans can execute accurate movements and generate precise muscle forces is very important for hand function. Target-tracking tasks or target matching tasks are often executed under combined visual and somatosensory feedback. When visual feedback is removed, subjects have to depend on their perception of force. The objective of the present study was to estimate the effects of aging on the perception of a pinch force produced by the thumb and index finger. In a first set of trials, young (n = 12, age = 25.3 +/- 2.4 years) and elderly (n = 12, age = 71.5 +/- 3.3 years) healthy individuals were asked to reproduce pinch forces which were equivalent to 5%, 20%, and 40% of their maximal pinch force (MPF). Prior to the execution of these trials, the subjects were familiarized with the force levels by matching targets displayed on a screen. They were then asked to reproduce each of these forces without any visual or verbal feedback. The results showed a larger error in the reproduced force for the elderly subjects when compared with the young adults. However, this larger error was mainly due to an initial overshoot in the force to be reproduced, followed by a gradual decrease towards the appropriate force. This transient overshoot was rarely seen in the performance of the younger subjects. In a second set of trials, the same subjects were asked to produce a pinch force of 5%, 20%, and 40% of MPF with 1 hand using visual feedback. They were also instructed to simultaneously apply a comparable pinch force with the other hand (without any feedback). For both young and older adults, the pinch forces produced by the 2 hands were the same. In addition, in both blocks of trials, hand dominance had no effects on the performance for all subjects. These results suggest that normal aging affects the production of force based on sensorimotor memory rather more than it affects comparative outputs from central descending commands. PMID- 15523527 TI - Movements generated by intraspinal microstimulation in the intermediate gray matter of the anesthetized, decerebrate, and spinal cat. AB - The intermediate laminae of the lumbosacral spinal cord are suggested to contain a small number of specialized neuronal circuits that form the basic elements of movement construction ("movement primitives"). Our aim was to study the properties and state dependence of these hypothesized circuits in comparison with movements elicited by direct nerve or muscle stimulation. Microwires for intraspinal microstimulation (ISMS) were implanted in intermediate laminae throughout the lumbosacral enlargement. Movement vectors evoked by ISMS were compared with those evoked by stimulation through muscle and nerve electrodes in cats that were anesthetized, then decerebrated, and finally spinalized. Similar movements could be evoked under anesthesia by ISMS and nerve and muscle stimulation, and these covered the full work space of the limb. ISMS-evoked movements were associated with the actions of nearby motoneuron pools. However, after decerebration and spinalization, ISMS-evoked movements were dominated by flexion, with few extensor movements. This indicates that the outputs of neuronal networks in the intermediate laminae depend significantly on descending input and on the state of the spinal cord. Frequently, the outputs also depended on stimulus intensity. These experiments suggest that interneuronal circuits in the intermediate and ventral regions of the spinal cord overlap and their function may be to process reflex and descending activity in a flexible manner for the activation of nearby motoneuron pools. PMID- 15523528 TI - Gait acts as a gate for reflexes from the foot. AB - During human gait, electrical stimulation of the foot elicits facilitatory P2 (medium latency) responses in TA (tibialis anterior) at the onset of the swing phase, while the same stimuli cause suppressive responses at the end of swing phase, along with facilitatory responses in antagonists. This phenomenon is called phase-dependent reflex reversal. The suppressive responses can be evoked from a variety of skin sites in the leg and from stimulation of some muscles such as rectus femoris (RF). This paper reviews the data on reflex reversal and adds new data on this topic, using a split-belt paradigm. So far, the reflex reversal in TA could only be studied for the onset and end phases of the step cycle, simply because suppression can only be demonstrated when there is background activity. Normally there are only 2 TA bursts in the step cycle, whereas TA is normally silent during most of the stance phase. To know what happens in the stance phase, one needs to have a means to evoke some background activity during the stance phase. For this purpose, new experiments were carried out in which subjects were asked to walk on a treadmill with a split-belt. When the subject was walking with unequal leg speeds, the walking pattern was adapted to a gait pattern resembling limping. The TA then remained active throughout most of the stance phase of the slow-moving leg, which was used as the primary support. This activity was a result of coactivation of agonistic and antagonistic leg muscles in the supporting leg, and represented one of the ways to stabilize the body. Electrical stimulation was given to a cutaneous nerve (sural) at the ankle at twice the perception threshold. Nine of the 12 subjects showed increased TA activity during stance phase while walking on split-belts, and 5 of them showed pronounced suppressions during the first part of stance when stimuli were given on the slow side. It was concluded that a TA suppressive pathway remains open throughout most of the stance phase in the majority of subjects. The suggestion was made that the TA suppression increases loading of the ankle plantar flexors during the loading phase of stance. PMID- 15523529 TI - Somatosensory paths proceeding to spinal cord and brain--centripetal and centrifugal control for human movement. AB - The flow of fast-conducting somatosensory information proceeding from the human leg, and entering sensorimotor control processes, is modulated according to the demands of limb movement. Both centripetal (proceeding in from sensory receptor discharge) and centrifugal (proceeding out from motor control centres) convergences can cause modulation, as seen in human, dog, and cat studies. Spinal H-reflexes appear to be strongly centripetally modulated in magnitude, as do initial somatosensory-evoked potentials recorded from the scalp following transmission in fast-conducting afferents from the leg. From the brain and from locomotor pattern-generators, there is also centrifugal control onto fast conducting somatosensory pathways from the leg, both serving spinal reflexes and ascending to the brain. One expression of the centrifugal control appears to be pattern-generator modulation of cutaneous reflexes. Centrifugal control also can be seen premovement, as spinal H-reflex facilitation. Further, it can be observed as reduction of reception at somatosensory cerebral cortex, when motor learning has occurred or when stimuli are less salient for the task. Fourteen research developments have been identified that involve the generalizability of effects, specific mechanisms, and somatosensory modulation in predictive control. PMID- 15523530 TI - Basal ganglia neural mechanisms of natural movement sequences. AB - Natural rodent grooming and other instinctive behavior serves as a natural model of complex movement sequences. Rodent grooming has syntactic (rule-driven) sequences and more random movement patterns. Both incorporate the same movements- only the serial structure differs. Recordings of neural activity in the dorsolateral striatum and the substantia nigra pars reticulata indicate preferential activation during syntactic sequences over more random sequences. Neurons that are responsive during syntactic grooming sequences are often unresponsive or have reverse activation profiles during kinematically similar movements that occur in flexible or random grooming sequences. Few neurons could be categorized as strictly movement related--instead they were activated only in the context of particular sequential patterns of movements. Particular sequential patterns included "syntactic chain" grooming sequences of paw, head, and body movements and also "warm-up" sequences, which consist of head and body/limb movements that precede locomotion after a period of quiet resting (Golani 1992). Activation during warm-up was less intense and less frequent than during grooming sequences, but both sequences activated neurons above baseline levels, and the same neurons sometimes responded to both sequences. The fact that striatal neurons code 2 natural sequences which are made up of different constituent movements suggests that the basal ganglia may have a generalized role in sequence control. The basal ganglia are modulated by the context of the sequence and may play an executive function in the complex natural patterns of sequenced behaviour. PMID- 15523531 TI - Rate-coding of spinal motoneurons with high-frequency magnetic stimulation of human motor cortex. AB - Rate-coding in spinal motoneurons was studied using high-frequency magnetic stimulation of the human motor cortex. The subject made a weak contraction to cause rhythmic (i.e., tonic) discharge of a single motor unit in flexor (or extensor) carpi radialis or tibialis anterior, while the motor cortical representation of that muscle was stimulated with brief trains of pulses from a Pyramid stimulator (4 Magstim units connected by 3 BiStim modules). An "m@n" stimulus train consisted of m number of pulses (1-4), with an interpulse interval (IPI) of n ms (1-6). Peristimulus time histograms were constructed for each stimulus condition of a given motor unit, and related to the average rectified surface electromyography (EMG) from that muscle. Surface EMG responses showed markedly more facilitation than single-pulse stimulation, with increasing numbers of pulses in the train; responses also tended to increase in magnitude for the longer IPI values (4 and 6 ms) tested. Motor-unit response probability increased in a manner comparable to that of surface EMG. In particular, motoneurons frequently responded twice to a given stimulus train. In addition to recruitment of new motor units, the increased surface EMG responses were, in part, a direct consequence of short-term rate-coding within the tonically discharging motoneuron. Our results suggest that human corticomotoneurons are capable of reliably following high-frequency magnetic stimulation rates, and that this activity pattern is carried over to the spinal motoneuron, enabling it to discharge at extremely high rates for brief periods of time, a pattern known to be optimal for force generation at the onset of a muscle contraction. PMID- 15523532 TI - Therapy of paretic arm in hemiplegic subjects augmented with a neural prosthesis: a cross-over study. AB - There are indications that both intensive exercise and electrical stimulation have a beneficial effect on arm function in post-stroke hemiplegic patients. We recommend the use of Functional Electrical Therapy (FET), which combines electrical stimulation of the paretic arm and intensive voluntary movement of the arm to exercise daily functions. FET was applied 30 min daily for 3 weeks. Forty one acute hemiplegics volunteered in the 18-months single blinded cross-over study (CoS). Nineteen patients (Group A) participated in FET during their acute hemiplegia, and 22 patients (Group B) participated in FET during their chronic phase of hemiplegia. Group B patients were controls during FET in acute hemiplegia, and Group A patients were controls during the FET in chronic hemiplegia. Thirty-two patients completed the study. The outcomes of the Upper Extremity Function Test (UEFT) were used to assess the ability of patients to functionally use objects, as were the Drawing Test (DT) (used to assess the coordination of the arm), the Modified Ashworth Scale, the range of movement, and the questionnaire estimating the patients' satisfaction with the usage of the paretic arm. Patients who participated in the FET during the acute phase of hemiplegia (Group A) reached functionality of the paretic arm, on average, in less than 6 weeks, and maintained this near-normal use of the arm and hand throughout the follow-up. The gains in all outcome scores were significantly larger in Group A after FET and at all follow-ups compared with the scores before the treatment. The gains in patients who participated in the FET in the chronic phase of hemiplegia (Group B) were measurable, yet not significant. The speed of recovery was larger during the period of the FET compared with the follow-up period. The gains in Group A were significantly larger compared with the gains in Group B. The FET greatly promotes the recovery of the paretic arm if applied during the acute phase of post-stroke hemiplegia. PMID- 15523533 TI - Encoding mechanisms for sensory neurons studied with a multielectrode array in the cat dorsal root ganglion. AB - Recent advances in microelectrode array technology now permit a direct examination of the way populations of sensory neurons encode information about a limb's position in space. To address this issue, we recorded nerve impulses from about 100 single units simultaneously in the L6 and L7 dorsal root ganglia (DRG) of the anesthetized cat. Movement sensors, placed near the hip, knee, ankle, and foot, recorded passive movements of the cat's limb while it was moved pseudo randomly. The firing rate of the neurons was correlated with the position of the limb in various coordinate systems. The firing rates were less correlated to the position of the foot in Cartesian coordinates (x, y) than in joint angular coordinates (hip, knee, ankle), or in polar coordinates. A model was developed in which position and its derivatives are encoded linearly, followed by a nonlinear spike-generating process. Adding the nonlinear portion significantly increased the correlations in all coordinate systems, and the full models were able to accurately predict the firing rates of various types of sensory neurons. The observed residual variability is captured by a simple stochastic model. Our results suggest that compact encoding models for primary afferents recorded at the DRG are well represented in polar coordinates, as has previously been suggested for the cortical and spinal representation of movement. This study illustrates how sensory receptors encode a sense of limb position, and it provides a general framework for modeling sensory encoding by populations of neurons. PMID- 15523534 TI - Motor unit firing rates during isometric voluntary contractions performed at different muscle lengths. AB - Firing rates of motor units and surface EMG were measured from the triceps brachii muscles of able-bodied subjects during brief submaximal and maximal isometric voluntary contractions made at 5 elbow joint angles that covered the entire physiological range of muscle lengths. Muscle activation at the longest, midlength, and shortest muscle lengths, measured by twitch occlusion, averaged 98%, 97%, and 93% respectively, with each subject able to achieve complete activation during some contractions. As expected, the strongest contractions were recorded at 90 degrees of elbow flexion. Mean motor unit firing rates and surface EMG increased with contraction intensity at each muscle length. For any given absolute contraction intensity, motor unit firing rates varied when muscle length was changed. However, mean motor unit firing rates were independent of muscle length when contractions were compared with the intensity of the maximal voluntary contraction (MVC) achieved at each joint angle. PMID- 15523535 TI - The effect of activity during early postnatal development on motor unit size. AB - At early stages of neuromuscular development, motor unit territory is expanded, with each muscle fibre being supplied by several axons. During postnatal development, some synapses are eliminated, motor unit size decreases, and the adult distribution of motor unit sizes emerges. This process depends on activity, since it proceeds more rapidly when the nerve is activated and is slower when activity is reduced. Here we studied whether, in addition to influencing the rate of retraction of motor unit territory, activity during the critical period of development affects the final outcome of the distribution of motor unit sizes. The sciatic nerve of 8- to 12-day-old rats was stimulated daily. One week later the tension of the extensor digitorum longus muscle and that of its individual motor units was recorded. The sizes of individual motor units were calculated and compared with those from animals that received no stimulation. The distribution of motor unit sizes from stimulated muscles was not significantly different from those from control muscles. Therefore, we conclude that although activity increases the rate at which motor units attain their adult size, it does not influence the final outcome of motor unit size distribution. PMID- 15523536 TI - Functional electrical stimulation using microstimulators to correct foot drop: a case study. AB - This paper presents a case study that tested the feasibility and efficacy of using injectable microstimulators (BIONs) in a functional electrical stimulation (FES) device to correct foot drop. Compared with surface stimulation of the common peroneal nerve, stimulation with BIONs provides more selective activation of specific muscles. For example, stimulation of the tibialis anterior (TA) and extensor digitorum longus (EDL) muscles with BIONs produces ankle flexion without excessive inversion or eversion of the foot (i.e., balanced flexion). Efficacy was assessed using a 3-dimensional motion analysis of the ankle and foot trajectories during walking with and without stimulation. Without stimulation, the toe on the affected leg drags across the ground. BION stimulation of the TA muscle and deep peroneal nerve (which innervates TA and EDL) elevates the foot such that the toe clears the ground by 3 cm, which is equivalent to the toe clearance in the less affected leg. The physiological cost index (PCI) measured effort during walking. The PCI equals the change in heart rate (from rest to activity) divided by the walking speed; units are beats per metre. The PCI is high without stimulation (2.29 +/- 0.37, mean +/- SD) and greatly reduced with surface (1.29 +/- 0.10) and BIONic stimulation (1.46 +/- 0.24). Also, walking speed increased from 9.4 +/- 0.4 m/min without stimulation to 19.6 +/- 2.0 m/min with surface and 17.8 +/- 0.7 m/min with BIONic stimulation. These results suggest that FES delivered by a BION is an alternative to surface stimulation and provides selective control of muscle activation. PMID- 15523537 TI - Direct and indirect assessment of gamma-motor firing patterns. AB - The study of the patterns of gamma-motor activity which accompany natural contractions has been long and difficult, and has not as yet led to general agreement. In this review we have simplified matters by considering the case of locomotion in the cat only, and we have avoided discussion of the various hypotheses which have been advanced to provide general schemes of gamma control for a wide range of movements. The development of the subject is shown to depend very much on devising ingenious methods applicable to reduced and intact animals. Direct recording from gamma-motoneurones has only been possible in reduced preparations, whereas indirect assessment of gamma activity from spindle afferent recordings was used in these and in intact animals. At this point in time, we still have no direct recordings from gamma-motoneurones in normally behaving animals, but those obtained in decerebrate animals show distinct patterns of modulation for static and dynamic types with particular temporal relation to the stepping movements. The spindle recordings in intact animals potentially provide the most important information, and the problems of interpretation, which have previously caused difficulties, are beginning to be solved through the insights obtained from the reduced preparations. PMID- 15523538 TI - Immunofluorescence revealed the presence of NHE-1 in the nuclear membranes of rat cardiomyocytes and isolated nuclei of human, rabbit, and rat aortic and liver tissues. AB - Using immunofluorescence and 3-dimensional confocal microscopy techniques, the present study was designed to verify if NHE-1 is present at the level of the nuclear membrane in cells that are known to express this type of exchanger. Nuclei were isolated from aortic tissues of adult human, rabbit, and rats, as well as from liver tissues of human fetus, and adult rabbit and rat. In addition, cultured ventricular cardiomyocytes were isolated from 2-week-old rat. Our results showed the presence of NHE-1 in isolated nuclei of aortic vascular smooth muscle and liver of human, rabbit, and rat. NHE-1 seems to be distributed throughout the isolated nucleus and more particularly at the level of the nuclear membranes. The relative fluorescence density of NHE-1 was significantly higher (p < 0.05) in isolated liver nuclei of human, when compared with those of rabbit and rat. However, in isolated nuclei of aortic vascular smooth muscle, the relative fluorescence density of NHE-1 was significantly (p < 0.001) higher in the rabbit when compared with human and rat. In cultured rat ventricular cardiomyocytes, NHE 1 fluorescent labeling could be easily seen throughout the cell, including the nucleus, and more particularly at both the sarcolemma and the nuclear membranes. In rat cardiomyocytes, the relative fluorescence density of NHE-1 of the sarcolemma membrane, including the cytosol, was significantly lower than that of the whole nucleus (including the nuclear envelope membranes). In conclusion, our results showed that NHE-1 is present at the nuclear membranes and in the nucleoplasm and its distribution and density may depend on cell type and species used. These results suggest that nuclear membranes' NHE-1 may play a role in the modulation of intranuclear pH. PMID- 15523539 TI - Aromatic hydrocarbon receptor status in the metabolism of xenobiotics under normal and pathophysiological conditions. AB - The aryl hydrocarbon receptor (AhR), a cytosolic protein belonging to the family of nuclear receptors, controls transcription of a wide range of structurally unrelated genes. To date, the most potent AhR ligand is 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD). Exposure to TCDD leads to a number of toxic effects, in particular to tumor promotion and immunosuppression. The function of AhR in cells and living organisms therefore seems to be of paramount importance. Its absence in AhR null mice, results in severe phenotypic abnormalities, such as liver half size with fibrosis, accumulation of retinoic acid and immune system insufficiency. An important role of AhR inheres in its transcriptional control of several biotransformation enzymes (CYP1A1/2,1B1). Hence AhR is the crucial factor in the regulation of xenobiotic metabolism. Under pathophysiological conditions, such as inflammation, the level and activity of the AhR target gene CYP1A is decreased. Thus it is likely, that mediators and/or products of inflammation affect AhR function. This review deals with the role of AhR in xenobiotic metabolism under normal and pathophysiological conditions, with respect to inflammation in particular. PMID- 15523540 TI - Khat habit and its health effect. A natural amphetamine. AB - Chewing the leaves of the khat shrub is common in certain countries of East Africa and Arabian Peninsula mainly Yemen. It has been established that a khat plant leaves contain an active psycho-stimulant substance known as cathinone that is similar in structure and pharmacological activity to amphetamine in affecting the CNS. Intoxication with khat is self-limiting but chronic consumption can cause certain health disturbances in the user and also lead to social and economic damage to the individual and the community. In recent years, several cases of intoxication have been observed outside the area of its use. In this view, the khat habit, its health effects and socioeconomic aspects are described with the political issue they imply. PMID- 15523541 TI - Determination of the glycaemic index of selected foods (white bread and cereal bars) in healthy persons. AB - The glycaemic index (GI) is a measure of the food power to raise blood glucose (B glucose) concentration after a meal. For healthy eating, foods with low GI are recommended. However, for many foods in the European Union the GI has not been defined yet. The aims of this prospective open-label study were: (1) to determine the GI of white bread and juicy cereal bars FIT (Usovsko, Czech Republic) by means of the glucometer Optium (Abbott/Medisense); (2) to compare the GI of tested foods determined in the morning and in the evening hours; (3) to compare the GI of tested foods in men and women and (4) to assess the variability of the GI. METHODS: To determine the GI, measured portions of food containing 50 g of carbohydrates were eaten by 11 healthy volunteers. B-glucose curves were constructed from B-glucose values at time 0, 15, 30, 45, 60, 60, 120 min after the meal. The GI was calculated by dividing the incremental area under the curve (IAUC) for the tested food by that for the standard food (IAUCS). In each volunteer each food was tested 5 times so that 5 GI's was obtained and the average was calculated. The GI for each tested food was calculated as the mean from the respective average GI's of the 11 volunteers. MS Excel and the statistical program SPSS v. 10.1 were used to analyze the data. RESULTS: (1) The mean values of the GI for white bread was 70.3 % and for juicy cereal bars was 101.0 %, as determined in a total of 139 tests in the whole group of 11 volunteers. There was a difference when comparing white bread vs. glucose (p = 0.012) and white bread vs. cereal bars (p = 0.026) but no difference between glucose and cereal bars. (2) There was no significant difference between the GI determined in the morning and in the evening hours either for the total of 139 tests or for the individual tested foods. (3) No significant difference could be seen between the GI in men and women when comparing glucose, cereal bars and white bread. (4) There was a wide variability of GI in all tested foods: the standard deviation of GI for white bread was 30.7 %, for juicy cereal bars 38.0 %. CONCLUSIONS: The GI's for white bread and juicy cereal bars were determined. There was no difference either between the GI values determined in the morning vs. the evening hours or between the values in men vs. women. The results show wide variability. An accurate standard method for the determination of GI needs to be defined, carefully used and re-evaluated to enable a comparison of the results with various methods of other working groups. PMID- 15523542 TI - Benefits of insulin aspart vs phosphate-buffered human regular insulin in persons with type 1 Diabetes treated by means of an insulin pump. AB - Absorption rates of phosphate buffered insulin analogs aspart and lispro prevail over regular human insulin. However, insulin aspart has not been widely used. The aim of this open controlled clinical study is to compare the metabolic effects of insulin aspart and phosphate buffered insulin when both are used in insulin pumps according to the identical algorithms. METHODS: Twenty one persons aged 39.9 +/- 2.89 (mean +/- SE) years (y) with type 1 diabetes mellitus duration of 17.9 +/- 2.21 y treated by an insulin pump for 4.3 +/- 0.53 y (at least 3 months), educated in self monitoring, entered the study. Mean plasma glucose, rates of hypo- and hyperglycaemias from the glucometer memory and other data from the first 256 +/- 19.97 days period with regular human insulin (check-up 1 and 2) and consequent 364 +/- 8.78 days long period with insulin aspart (check-up 3 and 4) were compared (paired t-test). Replacement of human regular insulin with insulin aspart after two check-ups was the only change in the treatment of diabetes. No special therapeutic education or training was made during the study. RESULTS: In persons with type 1 diabetes treated by an insulin pump with insulin aspart, despite the lower daily dose of insulin aspart vs human regular insulin, the HbA1c decreased; the frequency of hypo- and hyperglycaemias and the BMI did not change. CONCLUSIONS: Insulin analog aspart appears to be more effective for continuous subcutaneous insulin infusion than regular human insulin. PMID- 15523543 TI - A Continuous Glucose Monitoring System (CGMS) - a promising approach for improving metabolic control in persons with type 1 Diabetes mellitus treated by insulin pumps. AB - This pilot study deals with the possibilities of a Continuous Glucose Monitoring System (CGMS, Minimed- Medtronic) to optimize insulin substitution. Ten persons with type 1 diabetes mellitus treated by means of an insulin pump entered the study and eight of them completed the protocol. CGMS was introduced for a period of 5 days. The standard dinner (60 g of carbohydrates) and overnight fasting were designed to ensure standard night conditions in all persons in the study while maintaining their usual daily eating routine, physical exercise and assessment of prandial insulin boluses. The only adaptation of basal rates of insulin pump was performed on day 3. Comparison of the mean plasma glucose concentration (0:00 24:00 hrs) between day 2 (before adaptation) and day 4 (following adaptation) was made. An independent comparison of the mean plasma glucose concentration between the night from day 2 till day 3 (22:00-6:00 hrs) and the night from day 4 till day 5 (22:00-6:00 hrs) was performed. The mean plasma glucose investigated by means of CGMS improved in the 24-hour period in 5 out of 8 persons and in the night fasting period (22:00 to 6 hrs) in 6 out of 8 persons. The CGMS is a useful means for assessment of the effectiveness of basal rate and prandial insulin doses in persons with type 1 diabetes treated by means of an insulin pump. However, further studies are necessary to improve the algorithm for insulin substitution. PMID- 15523544 TI - Methods for retrospective assessment of an unusual behavior of a person with diabetes: ebrietas alcoholica and/or hypoglycaemia? AB - Drunkenness and/or hypoglycaemia are possible causes of unusual behavior in a person with diabetes. The aim of this study was to demonstrate different methods which were used in two contradictory reviews of experts for retrospective assessment of a patient's condition in the course of a car crash: An insulin treated driver caused a car accident resulting in injuries to two men. He was accused of the crime of damage to health as well as of driving under the influence of an addictive drug. One expert, having seen the police and medical reports, concluded the driver was influenced by alcohol at the time of the accident. The contradictory argument of another expert, based on his own thorough investigation including self monitoring and completed by a model experiment, resulted in the conclusion that the driver was probably influenced by hypoglycaemia. Thus, the methods of the second expert seem to be useful reasonable tools to assess the unusual behavior of a person with diabetes. PMID- 15523545 TI - Importance of guided fine needle aspiration cytology (FNAC) for the diagnostics of thyroid nodules - own experience. AB - The aim of our study was to assess the accuracy and limitations of ultrasound guided fine-needle aspiration cytology (FNAC) of thyroid nodules. METHODS: The usg-FNAC results of 245 patients with thyroid nodules, who afterwards underwent thyroid surgery or who died, and autopsies were carried out, and compared retrospectively with cytologic results. Patients with malignant cytologic conclusion without histological confirmation after surgery or autopsy were excluded from the study (9 persons). The usg-FNAC results were divided as follows: group 1: diagnosis of malignancy (n = 30), group 2: suspicion of malignancy (n = 28), group 3: benign (n = 126), group 4: inconclusive (n = 29). RESULTS: Assuming the cytologic results of group 1 and group 2 were interpreted as being malignant and those of group 3 as being benign, the sensitivity, specificity and accuracy of usg-FNAC were 90 %, 85 % and 86 % respectively. Comparing the cytologic conclusions between a group of patients with follicular lesions and a subgroup of other lesions a statistically significant difference (p < 0,01) between both subgroups using Fisher's test was found. Sensitivity, specificity and diagnostic accuracy in subgroup of follicular lesions were low (71 %, 63 %, 67 %), while in the subgroup of other lesions were high (94 %, 86 %, 88 %). FNAC can specify the nature of focal lesion with high sensitivity, specificity and diagnostic accuracy in the cases of non-follicular lesions. Histological evaluation is required to specification of the nature in cases in which cytology is indicative of follicular proliferation. PMID- 15523546 TI - Rewarming from severe accidental hypothermia with circulatory arrest. AB - This case report demonstrates successful cardiopulmonary and cerebral resuscitation (CPCR) of a young male explored 15 hours following a suicide attempt (carbamazepine intoxication) in deep hypothermia (19 degrees C) with circulatory arrest. An extracorporeal circuit was used to rewarm the patient's blood. Weaning from extracorporeal circulation (ECC) was successful and without complications as was recovery from multiorgan dysfunction, severe rhabdomyolysis and carbamazepine intoxication. An excellent outcome was achieved without any neurological deficit at the time of discharge from the hospital. PMID- 15523547 TI - Myocardial revascularisation in patients with severe left ventricular dysfunction. Early and midterm results. AB - AIMS: This retrospective study evaluates early and midterm results in patients with severe left ventricular dysfunction. TECHNIQUE: Ninety-seven consecutive patients with coronary artery disease and ejection fraction of left ventricle (LVEF) less or equal to 25 % underwent elective coronary artery bypass graft procedure between September 1998 and December 2001. Mean age at operation was 66 (47-86) years. The main symptoms were angina class III-IV Canadian Cardiovascular Society (CCS) in 78 patients (80 %) and dyspnoea class III-IV New York Heart Association (NYHA) in 43 patients (44 %). The major indications for surgery were severe angina, dyspnoea in 90 patients (93 %). Cardiac index (CI), echocardiography (ECHO), planimetry, end-diastolic pressure of left ventricle (LVEDP), end-diastolic diameter of left ventricle (LVEDd) were used to access left ventricular function preoperatively. ECHO was also used to access left ventricular function postoperatively. RESULTS: Early postoperative (30 days) mortality was 4.1 %. Complications had 25 patients (26 %) postoperatively. Two years survival was 95.7 %. Class III-IV CCS and/or NYHA had 17 patients (17.5 %) two years after surgery. LVEF (assessed by ECHO) improved from 23.1 % preoperatively to 36.0 % postoperatively (p < 0.05). CONCLUSION: The study showed that elective myocardial revascularisation in patients with severe left ventricular dysfunction and signs of myocardial viability is not necessarily associated with high operative mortality. It can be performed safely with respectable midterm survival in concordance with left ventricular function improvement, symptom relief and quality of life improvement. PMID- 15523548 TI - Cardiomed coronary flow meter for prevention of early occlusion in aortocoronary bypass grafting. AB - INTRODUCTION: The number of patients undergoing technically demanding off-pump myocardial revascularization is increasing, these days. Some researchers question the quality of the aortocoronary bypasses done on the beating heart and consequently their good patency. MATERIAL AND METHODS: 50 consecutive patients underwent off pump coronary surgery using sequential bypass technique, at our department. The quality of constructed grafts was evaluated using the CardioMed Trace System (CM4008, Medi-Stim As, Oslo, Norway) (TTFM). RESULTS: All sequential bypasses showed good per-operative quality with a mean fl ow of 69.4 ml/min. CONCLUSION: TTFM seems to be an effective tool for the per-operative aortocoronary bypass patency verification and should help to prevent early graft occlusion. ABBREVIATIONS: TTFM transit time flow meter. PMID- 15523549 TI - Short-term spectral analysis of heart rate variability during supine-standing supine test in patients with paroxysmal atrial fibrillation. AB - The aim of the study was to assess the sympathovagal balance in group of 27 patients without significant structural heart disease after an attack of atrial fibrillation. The investigation was performed using spectral analysis of heart rate variability during examination under conditions of different orthostatic loads in single phases, called the supine-standing- supine test. The findings were compared with a group of healthy persons. These revealed a significantly decreased total spectral power (430.7 vs 1558.0 ms(2) supine1; 477.6 vs 1042,5 ms(2) standing; 567.5 vs 1948.5 ms(2) supine2), and spectral power of the high frequency spectral component (140.8 vs 619.3 ms(2) supine1; 96.2 vs 203.3 ms(2) standing; 186.3 vs 739.4 ms(2) supine2) in the studied group of patients in comparison with the control group. PMID- 15523550 TI - Telmisartan in the treatment of hypertension in patients with chronic renal insufficiency. AB - The primary aim of this study was evaluation of the efficacy of telmisartan (angiotensin II receptor blocker- AT(1) blocker) on blood pressure in 10 patients with renal impairment in moderate or advanced stages of renal insufficiency and not dependent on haemodialysis. Its effect on proteinuria, renal function (represented by serum urea, creatinine, glomerular filtration), evaluation of overall therapy compliance in comparison with a previously prescribed angiotensin converting enzyme inhibitors (ACEI) were secondary aims. Considering the presence of left ventricle hypertrophy in all patients as a marker of hypertensive cardiopathy, the effect of telmisartan therapy on non-invasive cardiovascular parameters (ECG, echocardiography, and assessment of heart rate variability-HRV) was also evaluated. The study group involved 10 hypertensive patients (6 women, 4 men) with diabetic and non-diabetic renal impairment, proteinuria above 1 g/24 hours, hypertensive cardiopathy and intolerance of ACEI (cough). Telmisartan was added to their long-term antihypertensive combination therapy in a dose of 40 mg for the first 14 days, after which the dose increased to the maximal of 80 mg. The average initial daytime systolic blood pressure (SBP) was 149 +/- 19.7 mm Hg, average night-time SBP 145 +/- 23.0 mm Hg, average initial daytime diastolic BP (DBP) 90.6 +/- 2.5 mm Hg, night-time DBP 88.9 +/- 13.5 mm Hg. Average initial serum creatinine was 207.2 +/- 48.5 micromol/l, urea 15.1 +/- 4.4 mmol/l, GF 0.5 +/- 0.1 ml/s. Echocardiography revealed left ventricular (LV) hypertrophy with well preserved systolic and moderately impaired diastolic LV function. Also the HRV assessment revealed impaired neurovegetative (e.g. sympathovagal) balance. After 1 year of combination therapy with telmisartan, there was a clearly significant reduction in both SBP and DBP in both day and night-time (SBP daytime 149.6 vs.116.6 mm Hg, night-time 145.8 vs. 129.5 mm Hg; DBP daytime 90.6 vs. 83.5 mm Hg, night-time 88.9 vs. 79.3 mm Hg) and proteinuria (2.37 vs. 1.27 g/24 hour, p < 0.05). There were no significant changes in serum creatinine, urea values, and LV functions. On the other hand, further progression of the sympathovagal balance impairment was noted (continuing reduction of HRV in 9 from 10 patients), which can be described as the priority finding. The total compliance of telmisartan therapy was very good and without adverse clinical side effects. In conclusion - telmisartan reduces blood pressure and proteinuria safely and effectively in patients with various types of nephropathy in moderate or advanced stages of renal insufficiency. PMID- 15523551 TI - Our experiences with the treatment of periprosthetic fractures of femur. AB - In the statement authors want to draw attention to the possibilities of treatment of periprosthetic fractures of femur. They present their own experiences with the treatment of these fractures by using various types of internal and external fixation evaluated from 1996 to 2003. They present some less common types of internal fixation as e.g. fixation by clamp plates. PMID- 15523552 TI - Abdominal compartment syndrome in polytrauma. AB - Authors inform about the group of 8 patients with abdominal compartment syndrome (ACS) occurred as a complication in large blunt injury of abdominal cavity. To the ACS diagnose, the measurement of intracystic pressure is used routinely, whose values correlate fully with values of intraabdominal pressure (IAP). In case of increasing values of IAP over 25 mm Hg with positive clinical signs of ACS, authors indicate decompression laparotomy with temporary closing of abdominal cavity by sterile plastic foil or Ethizip. This preventive temporary laparostomy is recommended also in serious injuries of abdominal cavity in patients with fatal haemorrhage, treated by the method of staged laparotomy with tamponade of abdominal cavity and with massive blood and volume resuscitation. PMID- 15523553 TI - Peripheral venous hypertension after the creation of arteriovenous fistula for haemodialysis. AB - The function of an arteriovenous (av) fistula for haemodialysis may be complicated by manifestation of peripheral venous hypertension, which results from the arterial blood flow through the venous system into the periphery of the upper extremity. Its development is most typically caused by a proximal forearm av-fistula, as, in addition to the desirable arterialisation of the subcutaneous venous system of the arm, arterialisation of the venous system of the forearm and the hand may occur and possibly promote the development of venous hypertension, which may in the extreme result in gangrene of the fingers. Awareness of these problems as well as of the necessity of their surgical solution is essential for doctors dealing with haemodialysis. PMID- 15523554 TI - Peritoneovenous shunt - modification with the use of long saphenous vein. AB - The authors describe their own initial experience with saphenoperitoneal modification of the peritoneovenous shunt in intractable ascites solution. Their findings with this easy type of permanent ascites drainage using the "patient's own resources" are puzzling. PMID- 15523555 TI - Femoral - internal iliac bypass in aortoiliac aneurysms endovascular repair. AB - The authors describe femoral - internal iliac bypass creation to remove ischemic complications in aortoiliac aneurysm endovascular repair. Based on a good experience with bypass in 6 patients they recommend its preventive indication when both internal iliac arteries are overstented. PMID- 15523556 TI - Epidemiology and risk factors of kidney cancer. AB - This review article presents kidney cancer epidemiology as well as main environmental and life style risk factors. PMID- 15523557 TI - Macleya cordata and Prunella vulgaris in oral hygiene products - their efficacy in the control of gingivitis. AB - A double-blind, placebo-controlled clinical trial was performed to investigate the effectiveness of a herbal-based dentifrice in the control of gingivitis. Forty volunteers completed the 84-day study. All subjects were balanced for parameters measured - plaque index (PI), community periodontal index of treatment needs (CPITN) and papillary bleeding index (PBI). The dentifrice was effective in reducing symptoms of gingivitis as evaluated by the CPITN and PBI indexes. PMID- 15523558 TI - BMAL1 and CLOCK, two essential components of the circadian clock, are involved in glucose homeostasis. AB - Circadian timing is generated through a unique series of autoregulatory interactions termed the molecular clock. Behavioral rhythms subject to the molecular clock are well characterized. We demonstrate a role for Bmal1 and Clock in the regulation of glucose homeostasis. Inactivation of the known clock components Bmal1 (Mop3) and Clock suppress the diurnal variation in glucose and triglycerides. Gluconeogenesis is abolished by deletion of Bmal1 and is depressed in Clock mutants, but the counterregulatory response of corticosterone and glucagon to insulin-induced hypoglycaemia is retained. Furthermore, a high-fat diet modulates carbohydrate metabolism by amplifying circadian variation in glucose tolerance and insulin sensitivity, and mutation of Clock restores the chow-fed phenotype. Bmal1 and Clock, genes that function in the core molecular clock, exert profound control over recovery from insulin-induced hypoglycaemia. Furthermore, asynchronous dietary cues may modify glucose homeostasis via their interactions with peripheral molecular clocks. PMID- 15523560 TI - An unusual cause of urinary retention in the elderly patient. PMID- 15523559 TI - Components of coated vesicles and nuclear pore complexes share a common molecular architecture. AB - Numerous features distinguish prokaryotes from eukaryotes, chief among which are the distinctive internal membrane systems of eukaryotic cells. These membrane systems form elaborate compartments and vesicular trafficking pathways, and sequester the chromatin within the nuclear envelope. The nuclear pore complex is the portal that specifically mediates macromolecular trafficking across the nuclear envelope. Although it is generally understood that these internal membrane systems evolved from specialized invaginations of the prokaryotic plasma membrane, it is not clear how the nuclear pore complex could have evolved from organisms with no analogous transport system. Here we use computational and biochemical methods to perform a structural analysis of the seven proteins comprising the yNup84/vNup107-160 subcomplex, a core building block of the nuclear pore complex. Our analysis indicates that all seven proteins contain either a beta-propeller fold, an alpha-solenoid fold, or a distinctive arrangement of both, revealing close similarities between the structures comprising the yNup84/vNup107-160 subcomplex and those comprising the major types of vesicle coating complexes that maintain vesicular trafficking pathways. These similarities suggest a common evolutionary origin for nuclear pore complexes and coated vesicles in an early membrane-curving module that led to the formation of the internal membrane systems in modern eukaryotes. PMID- 15523561 TI - Induction of biomolecules in mature leaves of Terminalia arjuna due to feeding of Antheraea mylitta Drury. AB - Terminalia arjuna is an important food plant of the tasar silkworm, Antheraea mylitta Drury. In this study, we investigated the induction of biomolecules in mature leaves of these plants subjected to insect feeding. Increase in total tannin content, lipid peroxidation, and trypsin inhibitor activity have been observed in mature leaves damaged by the insects. The growth rate of Vth instar larvae of A. mylitta fed on previously damaged foliage reduced by 87.1%. Induction of biomolecules for defense mechanisms in relation to herbivore damage has been discussed. PMID- 15523562 TI - Spinal cord blood flow after ischemic preconditioning in a rat model of spinal cord ischemia. AB - Spinal cord blood flow after ischemic preconditioning is poorly characterized. This study is designed to evaluate spinal cord blood flow patterns in animals after acute ischemic preconditioning. Experiment 1: After a laminectomy and placement of a laser Doppler probe over the lumbar spinal cord to measure spinal cord blood flow, 16 male Sprague-Dawley rats were randomized into two groups: ischemic preconditioning (IPC, n = 8), and control (CTRL, n = 8). Rats in the CTRL and the IPC groups were subjected to 12 min of ischemia directly followed by 60 min of reperfusion. IPC rats received 3 min of IPC and 30 min of reperfusion prior to the 12-min insult period. Experiment 2: After instrumentation, the rats were randomized into three groups: control (CTRL, n = 7), ischemic preconditioning (IPC, n = 7), and time control (TC, n = 4). Rats in the CTRL and the IPC groups were subjected to the same ischemia and reperfusion protocol as above. The TC group was anesthetized for the same time period as the CTRL and the IPC groups, but had no ischemic intervention. Microspheres were injected at baseline and at 15 and 60 min into the final reperfusion. All rats were euthanized and tissue harvested for spinal cord blood flow analysis. In Experiment 1, there was a slight, significant difference in spinal cord blood flow during the ischemic period; however, this difference soon disappeared during reperfusion. In experiment 2, there was no difference in blood flow at any experimental time. The results of these experiments demonstrate that IPC slightly enhances blood flow to the spinal cord during ischemia; however, this effect is not sustained during the reperfusion period. PMID- 15523563 TI - High oxygen level in a soaking treatment improves early root and shoot development of black willow cuttings. AB - Black willow (Salix nigra) stem cuttings are commonly used to stabilize eroded streambanks with survival dependent on rapid development of adventitious roots to maintain plant water balance, absorb nutrients, and provide anchorage and support especially during flood and drought events. Soaking cuttings in water prior to planting increases survival and growth rates, but it is not known whether oxygen content in the soaking water affects the rate of early root and shoot initiation and growth. A laboratory experiment tested the hypothesis that cuttings treated with high oxygen (>95% saturation, 8.62 mg O2 l(-1)) soaking exhibit more rapid initiation and growth of roots and shoots than cuttings treated with low oxygen (<15% saturation, 1.24 mg O2 l(-1)) soaking and control (unsoaked). Root initiation was enhanced in both high and low O2 soaking treatments compared to control (100, 93, and 41%, respectively, n = 27). High O2 soaking led to greater root length than low O2 soaking during the fourth week after planting (26.5 and 12.3 cm on day 22; 27.7 and 19.1 cm on day 27, respectively). Shoot growth was greater in high O2 compared to low O2 soaking on days 36 and 56 after planting (9.3 and 6.3 cm on day 36, 10.7 and 7.2 cm on day 56, respectively). Shoot and root biomass production was stimulated in both soaking treatments, with 200% more biomass production by day 59 compared to control. Results of this study demonstrated that a high oxygen soaking treatment has potential for improving early root and shoot growth, and survival in willow cuttings planted at riparian restoration sites. PMID- 15523564 TI - Pilonidal sinus of the glans penis associated with actinomyces case reports and review of literature. AB - Pilonidal sinus is a well-recognized condition that occurs most commonly in the sacrococcygeal area of younger men. It is hypothesized to be an acquired chronic inflammation condition due mainly to hair trapped beneath the surface. A pilonidal sinus in the sacrococcygeal region is associated with recurrent infection, abscess formation, cellulitis, fistulae, and rarely, squamous cell carcinoma. A pilonidal sinus of the penis is a rare entity. The association of a penile pilonidal cyst and Actinomyces is even more uncommon with only three cases reported previously. Two cases of pilonidal sinus are reported in this paper. One of the cases was associated with actinomycosis. Pilonidal sinus of the penis should be considered in the clinical and pathological differential diagnosis and has to be distinguished from balanoposthitis, epidermal cyst, and carcinoma. The knowledge about possible association with actinomycosis is important to ensure early treatment. PMID- 15523565 TI - Clinical holistic medicine: metastatic cancer. AB - We believe that the consciousness-based/holistic medical toolbox has a serious additional offer to cancer patients and, as a consequence, designed a treatment for the patient with metastasized cancer. From a holistic perspective, cancer can be understood as a simple disturbance of the cells, arising from the tissue holding on to a trauma with strong emotional content. This is called "a blockage", where the function of the cells is allocated from their original function in the tissue to a function of holding emotions. We hope to be able not only to improve the quality of life, but also to improve survival and in some cases even induce spontaneous remission of the metastasized cancer. This paper describes how work with a patient with metastasized cancer can be done in the holistic clinical practice in 14 days on an individual basis, helping the patient to recover her human character, purpose of life, coherence, and will to live, thus improving quality of life and possibly also survival time. The holistic therapeutic work includes (1) teaching existential theory, (2) working with life perspective and philosophy of life, (3) helping the patient to acknowledge the state of the disease and the feelings connected to it, and finally (4) getting the patient into the holistic state of healing: (a) feeling old repressed emotions, (b) understanding why she got sick from a holistic point of view, and finally (c) letting go of the negative beliefs and decisions that made her sick according to the holistic theory of nongenetic diseases. The theory of the human character, the quality of life theories, the holistic theory of cancer, the holistic process theory of healing, the theory of (Antonovsky) coherence, and the life mission theory are the most important theories for the patient to find hope and mobilize the will to fight the cancer and survive. The patient went through the following phases: (1) finding the purpose of life and hidden resources; (2) confronting denial; (3) taking responsibility for being very ill; (4) severe existential crises with no wish to live while still fighting; (5) integration of many repressed feelings and negative decisions thus rehabilitating character; (6) confronting lack of intimacy and trust in others and this way rehabilitating the ability to love; (7) rehabilitating the will to live, breaking through and falling in love with life; (8) assuming responsibility for the social relations; and sometimes (9) quality of life is improved radically with indications of spontaneous remission of the liver tumors. PMID- 15523566 TI - [Reusing single-use medical devices]. PMID- 15523568 TI - [In R&D projects... who is vouching for the evaluator?]. PMID- 15523567 TI - Emergence of Serratia marcescens as an ocular surface pathogen. PMID- 15523569 TI - [Chronic carriers of pathogen conjunctival bacteria. Possible risks in cataract surgery]. AB - PURPOSE: To ascertain the frequency and characteristics of chronic carriers of conjunctival pathogen bacteria among patients undergoing cataract surgery in our hospital, to allow the design of studies of their postsurgical endophthalmitis risk. METHODS: Retrospective study of the preoperative conjunctival flora of 784 patients operated on for cataracts in both eyes, in two operations separated by 213 days (SD 170), from November 1993 to December 1997. Results of both cultures for each patient were obtained from the Laboratory cataract preoperative database, by means of an auxiliary utility in dBASE-III-PLUS program. The preoperative bacteria in both surgeries were compared and the patients having the same pathogen bacteria (all except Staphylococcus coagulase negative and Corynebacterium sp.) were identified. The mean values and standard deviations were calculated using Epiinfo 6.04, and the Chi2 test was carried out using Excel 97. RESULTS: The conjunctival flora stemming from the first preoperative culture of the 784 patients were statistically similar to those of our reference population. The pathogen bacteria decreased in the second preoperative culture; especially the Streptococcus Pneumoniae and Haemophilus sp. frequencies (p<0.05). The same pathogen was isolated in both cultures of 31 patients among the 784 who entered the study, Staphylococcus Aureus and Proteus Mirabilis being the most frequent colonizers. The mean age of those carriers was higher than the mean age of the reference population (p<0.01). CONCLUSIONS: 3.95% of our patients awaiting cataract surgery are usual conjunctival carriers of certain pathogen bacteria; this finding is associated with the age of the patients and possibly with some pre-existing diseases. PMID- 15523570 TI - [Neoplasms of the caruncle. Clinicopathologic study of 40 cases]. AB - PURPOSE: To determine the type and frequency of the neoplasms on the lachrymal caruncle and to compare the results with other published case series. METHODS: Retrospective study of forty cases of lesions of the lachrymal caruncle, surgically treated during a 13-year period (1991-2003) are presented. There were 19 females and 21 males. The mean age was 42.1, SD 19.5 years. RESULTS: Ten different histopathologic types of lesions were found. The most frequent lesion was the nevus (40%), and the second, the papilloma (30%). Thirty-seven neoplasms (92.5%) were benign, 2 (5%) premalignant and 1 (2.5%) malignant. The clinical diagnosis was accurate in 87.5% of the cases. CONCLUSION: In this case series, we confirm the low frequency of caruncular neoplasms as well as the wide variety of histopathologic types found in this location. There was a wide range of ages and we did not find any evident sex predominance. Malignant lesions were uncommon, with only one case of basal cell carcinoma. Our results are similar to those of other case series. PMID- 15523571 TI - [Treatment of nasolacrimal duct obstruction with Song's stent. Long term efficacy]. AB - PURPOSE: To evaluate the long-term efficacy (5 years) of Song's stent in the treatment of naso-lachrymal duct obstruction. MATERIALS AND METHODS: We examined Song's stents installed in 64 eyes from 62 patients with naso-lachrymal duct obstruction (NLD). Of the 64, 48 were women and 14 men of mean age 60 SD 13.17 years. Obstructions of right side 62.5% and left side 37.5%, and location in dacriocystography: sac-NLD junction 82.8%, distant NLD 10.9%, medium NLD 3.1% and both proximal and complete NLD 1.6%. CLINICAL PRESENTATION: chronic epiphora 96.8%, acute dacriocystitis 67.2%, purulent discharge 25% and mucocele 17.2%. Mean follow-up period was 25.23 months. RESULTS: 13 stents were removed and 2 changed during the follow-up. 67.2% were patent at the end of the study; epiphora reduced to 32.8%, number of acute dacryocystitis, purulent discharge and mucocele to 1.6%. Subjective improvement was found in 73.5%. Surgical complications included epistaxis (15.6%) and pain (14.1%), and post-operative complications included reobstruction in 22 cases within a mean time of 8.6 months. CONCLUSIONS: Song's stent is a quick and useful option in treatment of epiphora. Despite not being efficient in all cases, it leads to improvement of symptoms and avoidance of surgery in the majority of cases. However, it requires periodical washes to maintain its patency. PMID- 15523572 TI - [Central serous chorioretinopathy versus sympathetic ophthalmia. Case report]. AB - CLINICAL CASE: A 29 year old patient with a history of sutured corneal wound treated with oral antibiotics and steroids. After stopping steroid treatment, he referred reduced visual acuity in the non-traumatized eye. Previous treatment was reintroduced and he was referred to our hospital diagnosed with sympathetic ophthalmia. Bilateral macular folds, white and yellowish lesions, and subretinal fluid were found. DISCUSSION: The association of stress, anxious personality, and steroid use can favor atypical presentations of central serous chorioretinopathy. PMID- 15523573 TI - [Symptomatic ciliary body melanocytoma: melanocytomalytic glaucoma]. AB - CASE REPORT: A 33 year-old female with an asymptomatic pigmented mass in the iridocorneal angle of her right eye, arising from the ciliary body is presented. Ciliary body melanocytoma was suspected and conservative management recommended. After 36 months of follow-up the patient developed pain, inflammatory reaction and uncontrollable ocular hypertension, which was diagnosed as melanocytomalytic glaucoma. Tumor was removed by external iridecyclectomy and the histopathologic findings revealed necrotic melanocytoma. DISCUSSION: Ciliary body melanocytoma is a rare benign pigmented tumor that may present extension to the anterior chamber. Differential diagnosis mainly includes ciliary body melanoma, which carries a different prognosis and treatment. PMID- 15523574 TI - [Orbit xanthogranulomatosis. Erdheim-Chester disease]. AB - CASE REPORT: A patient was studied because of upper lid bilateral edema and xanthelasmae-like lesions after three years of evolution. During the ophthalmologic examination orange-yellowish lesions and two symmetrical tumours were observed on the temporal part of both upper lids. Corticoid-therapy was undertaken which reduced the size of the tumours, however the size increased again after the discontinuation of treatment. A biopsy was performed and lid xanthogranulomatosis was diagnosed. Other systemic examinations were normal. DISCUSSION: Erdheim-Chester disease is a xanthogranulomatosis that can affect ocular and periorbital structures. Combination of xanthelasmae-like lesions and bilateral orbital masses should make us consider this process and try to locate any associated systemic conditions. PMID- 15523575 TI - [Oideo: a case of quack ophthalmology?]. PMID- 15523576 TI - [Wiley Hardeman Post, another one-eyed aviator]. PMID- 15523577 TI - [CGRP-receptor antagonists for the treatment of acute migraine: interesting option or therapeutic breakthrough?]. PMID- 15523578 TI - [Innovative treatment of acute migraine pain with CGRP receptor antagonists]. AB - Neurogenic inflammation caused by the release of vasoactive neuropeptides such as calcitonin gene-related peptide (CGRP) from nociceptive perivascular nerve fibres has been implicated in the development of acute migraine pain. It has recently been demonstrated that the CGRP receptor antagonist BIBN 4096 BS significantly reduces acute pain in migraine. This therapeutic efficacy provides further evidence for a critical role of CGRP in the pathophysiology of migraine. Given the apparent absence of vasoconstrictor activity both in meningeal and extracranial vessels, BIBN 4096 BS may provide an alternative for the treatment of migraine in those patients where triptans and ergotamine derivatives are contraindicated due to their side effects. PMID- 15523579 TI - [Injuries to car passengers protected by air bags]. AB - OBJECTIVE: The air bag, like the seatbelt, is a further development of the inside protection of motorcar passengers. However, the airbag has also been made responsible for severe internal injuries. METHODS: In a retrospective case control study, 394 accidents in which the air bag was released were analysed. At least medium severe injuries (Maximum Abbreviated Injury Scale: MAIS > or = 2) occurred in 69 cases. Three different patterns of injury were distinguished depending on the level of difficulty of diagnosis by the emergency physician. Damage to the vehicles was scored in five intensities or damage grades. RESULTS: Thoracic injury was most frequently diagnosed in the patients (in 61.5 % of cases), followed by injuries to the lower (50.8 %) and upper extremities (47.7 %). Single injuries with a grade of severity of 2 (MAIS) predominated (59.7 %). In most of the cases the injury was easy to diagnose (64.6 %) because of external signs, in 24.6 % internal injuries were assumed and in only 10.8 % were there no sings of damage to body cavities. Most frequent were occult injuries in the thoracic region (100 %) and in the abdomen (74.4 %). However, occult injuries did not always conform to the grade of deformation to the vehicle, since in 66.7 % the grade of damage was 3. This was not true for the remaining types of injury because external injuries increased with the grade of damage to the vehicle. CONCLUSION: After the release of the air bag, occult injuries of the body cavities have to be expected, even if there are no signs of external injury. Women under 35 years of age are particularly endangered. There exists no minimum velocity for the occurrence of injuries to the body cavities because harm can simply be a result of the release of the air bag. PMID- 15523580 TI - [Rapid sequence induction in prehospital emergency medicine: is it safe?]. AB - OBJECTIVE: The use of 0,9 mg/kg Rocuronium allows endotracheal intubation within 60 seconds and therefore might be an alternative to succinylcholine for rapid sequence induction. We investigated the use of high-dose Rocuronium for RSI in prehospital emergency medicine. METHODS: Rocuronium was used in two physician staffed units of emergency medical service for RSI in 79 patients aged 4 - 81 yrs (mean 46 yrs). Anesthesia was induced with etomidate/fentanyl or ketamine and 1 mg/kg of rocuronium. Recorded data were number of intubations, time interval application rocuronium-intubation, rating of intubation conditions and side effects. RESULTS: In 75 of 79 (94,9 %) patients intubation could be performed at the first attempt. In three (3,8 %) patients a second attempt and in one pt. a third attempt was necessary. In 78 (98,7 %) patients intubation could be performed within 60 seconds, in one patient intubation was completed within three minutes. The intubation conditions were estimated as excellent (n = 69; 87,3 %) or good (n = 10; 12,7 %) in all patients. No specific side effects could be observed. CONCLUSIONS: The use of muscle relaxants improves the intubation conditions and may help to avoid unnecessary high doses of anesthetics in hemodynamic unstable emergency patients. This study shows that rocuronium might be an alternative to succinylcholine for RSI also in prehospital emergency medicine, if succinylcholine is contraindicated. PMID- 15523581 TI - [Excessive blood loss after abdominal hysterectomy -- use of recombinant factor VIIa]. AB - MEDICAL HISTORY: A 38-yr-old woman suffered from excessive blood loss after elective removal of the uterus because of a leiomyoma. The surgical attempt to stop the bleeding failed. The laboratory evaluation before surgery showed normal coagulation parameters (aPTT 23.4 sec, TPZ > 100 %, platelet count 267 000/microl). DEVELOPMENT: Despite treatment with the plasminogen activator inhibitor Tranexamic Acid (2 x 500 mg) and 10 units of concentrated platelets as well as 31 units of fresh frozen plasma and 31 units of red blood cell concentrates the bleeding continued. Reaching an cumulative blood loss of 15 000 ml the patient developed ventricular fibrillation necessitating cardiac resuscitation including defibrillation and application of epinephrine 7 mg. At this point systolic blood pressure decreased from 90 to 70 mm Hg, the core temperature was 31,9 degrees C. After successful resuscitation (systolic blood pressure 90 mm Hg after 15 min) the bleeding started again with a blood loss of 2000 ml requiring 3 units of fresh frozen plasma and 3 red cell concentrates Use of recombinant activated Factor VII (r.FVIIa): 6 mg (300 KiU) r.FVIIa have been applied (92 microg/kgKG [4.6 KiE/kg]). Within 10 minutes the bleeding stopped. More transfusions were not necessary. The patient left the intensive care unit 8 days after the operation without neurological deficit. CONCLUSION: The use of recombinant Factor VIIa may stop excessive haemorrhage, even if the cause is unknown. PMID- 15523582 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Intraoperative blood salvage with blood irradiation -- from an anaesthesiological point of view]. AB - The requirement for intraoperative blood salvage (IBS) in cancer surgery stems from the high transfusion rate, the unfavourable effects of an anaemia, and the impact of transfusion risks like immunomodulation in tumor patients. The advantages of IBS are availability, the low waste rate, and the excellent quality of this autologous, unstored blood. The only effective elimination of the risk of tumor cell dissemination after retransfusion of wound blood is achieved by blood irradiation. The combination of the established methods of IBS and blood irradiation is practical, and allows a very efficient saving of blood. For Jehovah's witnesses it may open the possibility for tumor surgery. From an anaesthesiological point of view it is part of the therapy, but any context that puts it compatible to medicolegal regulations is welcome. For him as the one responsible for the therapy of intraoperative blood loss it represents the safest and best blood for an optimal hemotherapy in tumor patients. In addition, first data indicate a better outcome of these patients. PMID- 15523583 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Blood irradiation for intraoperative autotransfusion in cancer surgery -- the view of transfusion medicine]. PMID- 15523584 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Amendment to the transfusion act: easing of legal provisions on salvaged blood preparations]. PMID- 15523585 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Autologous transfusion in children -- blood saving techniques]. AB - It is uncontested that blood-saving strategies should also be applied in children. However, in the past, blood-saving techniques saw limited use in children, although they are well-established in adults. This is due to technical and methodical problems, but also to the physiologically limited compensatory mechanisms for diminished oxygen delivery in this age group. For this reason, the various blood saving-techniques cannot be universally applied to all age groups. During the first year of life most perioperative techniques are of only limited benefit: even only a small amount of blood loss in relation to total blood volume makes a transfusion of allogeneic blood necessary, which is far before enough shed blood can be recovered for retransfusion. Preoperative autologous donation of blood calls for a high degree of cooperation by the child and the parents an is equally demanding in terms of organization and skills. Therefore, this procedure is used mainly in school-age children and in adolescents. Intraoperative blood salvage, by contrast, is already worthwhile in children from age one year with an expected blood loss of 20 % of blood volume. Acute normovolemic hemodilution and deliberate hypotension should be recommended only in an age group where cardiovascular compensatory mechanisms are given. Supporting procedures aimed at avoiding the need for allogeneic blood include perioperative administration of erythropoietin, iron supplementation, blood saving surgical techniques and careful hemostasis. These prerequisites, the combination of various techniques, as well as the definition of an age-specific low transfusion trigger have contributed to a marked decrease in the need for allogeneic blood products in children in recent years. PMID- 15523586 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Postoperative shed mediastinal blood retransfusion -- pro]. PMID- 15523587 TI - [Autologous transfusion -- from euphoria to reason: clinical practice based on scientific knowledge (Part II). Autologous direct re-transfusion -- contra]. PMID- 15523588 TI - Undue increase in insulin resistance during pregnancy may manifest pregnancy induced hypertension and gestational diabetes. PMID- 15523589 TI - Increase of PTEN gene expression in insulin resistance. AB - Phosphatase and tensin homologue deleted from chromosome ten (PTEN) has recently been characterized as a regulator of insulin sensitivity in the insulin target tissue. However, whether PTEN gene expression is changed in insulin resistance remains unclear. We observed that both the mRNA and protein level of PTEN in soleus muscle isolated from the obese Zucker rats (Fa/Fa) were increased compared to the age-matched lean group. Similarly, both the mRNA and protein level of PTEN in soleus muscle of the fructose-fed lean Zucker rats (Fa/Fa) showing the higher glucose-insulin index were higher than that of the regular chow fed group. These results suggest that increase of PTEN gene expression seems to be related to the development of insulin resistance. PMID- 15523590 TI - Short-term fasting and lipolytic activity in rat adipocytes. AB - The aim of this experiment was to study the influence of 18-hour food deprivation on basal and stimulated lipolysis in adipocytes obtained from young male Wistar rats. Fat cells from fed and fasted rats were isolated from the epididymal adipose tissue by collagenase digestion. Adipocytes were incubated in Krebs Ringer buffer (pH 7.4, 37 degrees C) without agents affecting lipolysis and with different lipolytic stimulators (epinephrine, forskolin, dibutyryl-cAMP, theophylline, DPCPX, amrinone) or inhibitors (PIA, H-89, insulin). After 60 min of incubation, glycerol and, in some cases, also fatty acids released from adipocytes to the incubation medium were determined. Basal lipolysis was substantially potentiated in cells of fasted rats in comparison to adipocytes isolated from fed animals. The inhibition of protein kinase A activity by H-89 partially suppressed lipolysis in both groups of adipocytes, but did not eliminate this difference. The agonist of adenosine A (1) receptor also did not suppress fasting-enhanced basal lipolysis. The epinephrine-induced triglyceride breakdown was also enhanced by fasting. Similarly, the direct activation of adenylyl cyclase by forskolin or protein kinase A by dibutyryl-cAMP resulted in a higher lipolytic response in cells derived from fasted animals. These results indicate that the fasting-induced rise in lipolysis results predominantly from changes in the lipolytic cascade downstream from protein kinase A. The antagonism of the adenosine A (1) receptor and the inhibition of cAMP phosphodiesterase also induced lipolysis, which was potentiated by food deprivation. Moreover, the rise in basal and epinephrine-stimulated lipolysis in adipocytes of fasted rats was shown to be associated with a diminished non-esterified fatty acids/glycerol molar ratio. This effect was presumably due to increased re-esterification of triglyceride-derived fatty acids in cells of fasted rats. Comparing fed and fasted rats for the antilipolytic effect of insulin in adipocytes revealed that short-term food deprivation resulted in a substantial deterioration of the ability of insulin to suppress epinephrine-induced lipolysis. PMID- 15523591 TI - Testosterone increases osteoprotegerin mRNA expression in mouse osteoblast cells. AB - The role that androgens play in the regulation of bone metabolism has been substantiated in animals and humans. We previously demonstrated that testosterone inhibits osteoclast differentiation stimulated by parathyroid hormone through the androgen receptor in mouse bone-cell cultures. However, the details of this mechanism are still unknown. The present study was aimed at examining whether testosterone would affect the mRNA levels of osteoprotegerin (OPG) and receptor activator of Nf kappa B ligand (RANKL) in mouse bone-cell cultures as well as mouse osteoblastic cell-line, MC3T3-E1 cells by employing semi-quantitative RT PCR. Testosterone increased OPG mRNA expression in both mouse bone-cell cultures and MC3T3-E1 cells. 10-8 M PTH-(1-34) as well as 10-8M 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] inhibited OPG mRNA expression in mouse bone cells. 10-8 M testosterone antagonized OPG mRNA expression inhibited by 10-8 M PTH-(1-34), but failed to affect OPG mRNA expression inhibited by 10-8 M 1,25(OH)2D3. 10-8 M alpha-dehydrotestosterone, a non-aromatizable androgen, increased OPG mRNA expression. On the other hand, testosterone did not affect RANKL mRNA expression in MC3T3-E1 or mouse bone cells. In conclusion, the present study demonstrated that testosterone increased OPG mRNA expression in mouse bone-cell cultures and the osteoblastic cell line. These effects are likely to take place through the androgen receptor. PMID- 15523592 TI - 3,5-diiodothyronine mimics the effect of triiodothyronine on insulin-like growth factor binding protein-4 expression in cultured rat hepatocytes. AB - We have previously demonstrated that triiodothyronine (T(3)) stimulates hepatic IGFBP-4 expression in rats. Since there is evidence that some of the genes whose expression is regulated by T(3) are also sensitive to 3,5-diiodothyronine (T(2)), we used the adult rat hepatocyte model in primary cultures directly exposed to T(2) to evaluate insulin-like growth factor binding protein-4 (IGFBP-4) expression by Northern and Ligand blot analyses in this study. Our results demonstrate that T(2), like T(3), is able to enhance IGFBP-4 mRNA and protein after 12-24 h of incubation. The potency of the two iodothyronines is comparable as judged by dose-dependence experiments. The T(2)-induced IGFBP-4 increase is independent from ongoing protein synthesis but dependent on active transcription. Since T(3) and T(2) do not affect IGF-I production, it appears that the iodothyronines affect the hepatic IGF system at the IGFBP level. Our data, demonstrating that T(2) mimics the stimulatory effect of T(3) on IGFBP-4 expression by rat hepatocytes, allow us to include IGFBP-4 gene among the genes regulated by the two iodothyronines. PMID- 15523593 TI - Met326Ile aminoacid polymorphism in the human p85 alpha gene has no major impact on early insulin signaling in type 2 diabetes. AB - Class I alpha phosphatidylinositol (PI) 3-kinase is an important enzyme in the early insulin signaling cascade, and plays a key role in insulin-mediated glucose transport. Despite extensive investigation, the genes responsible for the development of the common forms of type 2 diabetes remain unknown. This study was performed to identify variants in the coding region of p85 alpha, the regulatory subunit of PI 3-kinase. Fibroblasts from skin biopsies from type 2 diabetics and controls were established to address this issue. P85 alpha cDNA was sequenced, and a single point mutation at codon 326 was found. This mutation resulted in a homozygous missense amino acid change Met --> Ile in one subject with type 2 diabetes and heterozygous variant in two other diabetic patients and one with severe insulin resistance. Interestingly, those patients revealed an impaired insulin-mediated insulin receptor substrate (IRS)-1 binding to p85 alpha without any alteration in IRS-2/p85 alpha association. Furthermore, IRS-1, IRS-2, p85 alpha and MAPK protein contents were not significantly changed, and neither were MAPK or Akt phosphorylation. We conclude from our data that this variant may have only minor impact on signaling events; however, in combination with variants in other genes encoding signaling proteins, this may have a functional impact on early insulin signaling. PMID- 15523594 TI - Lipid peroxidation at various estradiol concentrations in human circulation during ovarian stimulation with exogenous gonadotropins. AB - The aim of our study was to investigate the correlation between serum malondialdehyde levels and serum estradiol concentrations in healthy human female subjects. Nine hundred and fifty-five blood samples, from infertile women undergoing controlled ovarian hyperstimulation treatment with recombinant follicle-stimulating hormone, were collected for estradiol and malondialdehyde measurements. Five groups were formed according to serum estradiol levels: Group I (< 50 pg/ml), group II (50 - 299 pg/ml), group III (300-999 pg/ml), group IV (1000-1999 pg/ml) and group V (> or = 2000 pg/ml). One-way analysis of variance was used for comparisons. Mean malondialdehyde concentrations were 1.74 +/- 0.24 mmol/ml (group I), 1.53 +/- 0.20 mmol/ml (group II), 1.69 +/- 0.24 mmol/ml (group III), 1.77 +/- 0.21 mmol/ml (group IV) and 1.86 +/- 0.20 mmol/ml (group V), respectively. Mean serum malondialdehyde level at physiological estradiol concentrations (50-199 pg/ml, group II) was significantly (p < 0.01) lower than the mean malondialdehyde levels in other groups. Mean malondialdehyde concentrations among the remaining groups did not significantly differ. Our findings suggest that in vivo lipid peroxidation might be increased when circulating estradiol concentrations are below (< 50 pg/ml) or above (> 300 pg/ml) the physiological limits. High blood estradiol levels in human female subjects during ovarian stimulation with exogenous gonadotropins could be associated with increased serum malondialdehyde concentrations. PMID- 15523595 TI - Insulin-like growth factor-I stimulates acromegaly-like specific mandibular enlargement in rats. AB - To help us investigate the time course of mandibular enlargement in acromegaly or acrogiantism to determine the most suitable period for occlusal treatment in this disease, our aim was to develop a rat model of acromegaly (acrogiantism). In this study, prominent mandibular enlargement was induced by continuous subcutaneous infusion of human recombinant insulin-like growth factor-I (IGF-I) (640 microg/day) in 10-week-old male rats for 4 weeks (n = 6); the control sham operated group was injected with saline alone (n = 6). Circulating human IGF-I was clearly detectable in the IGF-I group during the four-week administration period, while endogenous rat IGF-I levels decreased. Total IGF-I (human + rat) increased significantly during administration, returning to control levels afterwards. The length of every bone examined (mandible, maxilla, and femur) showed a significant increase compared to control rats, especially the mandible. Although the mandible did not continue to grow after discontinuation of IGF-I administration, it did not return to control size, unlike the maxilla and femur, and disharmonious jaw size (between maxilla and mandible) persisted even after circulating IGF-I levels normalized. These findings in our rat model suggest that mandibular occlusal treatment should only be considered for acromegalic (acrogiantic) patients after serum IGF-I levels have normalized and bone growth has ceased. PMID- 15523596 TI - Identification of variables influencing resistin serum levels in patients with type 1 and type 2 diabetes mellitus. AB - BACKGROUND: Resistin, a peptide hormone, has been discussed controversially as a missing link between obesity and insulin resistance. In contrast to resistin mRNA expression in adipose tissue, data on human serum levels in obesity and diabetes mellitus is scarce. The physiological range of serum resistin levels, reference values or adjusted percentiles have not yet been determined, making the interpretation of serum resistin concentrations quite difficult. METHODS: Resistin serum concentrations were measured systematically by ELISA in 216 healthy controls, 555 patients with type 2 diabetes and 114 patients with type 1 diabetes. Mean values, median, and range were determined, and BMI-, gender-, and disease-adapted percentiles were calculated for all subgroups. RESULTS: Age and gender did not have any influence on resistin levels. BMI and resistin levels were positively correlated in healthy controls (p = 0.02), albeit with a weak correlation coefficient. This correlation was absent in patients with type 1 and type 2 diabetes. In both genders, healthy controls had significantly higher resistin levels than patients with type 1 and type 2 diabetes (7.9 +/- 0.2 ng/ml vs. 5.7 +/- 0.2 ng/ml and 5.5 +/- 0.1 ng/ml, respectively; p < 0.0001). There was no correlation between resistin levels and occurrence of diabetic retinopathy or nephropathy. CONCLUSIONS: Serum resistin levels can be measured by ELISA over a broad range from 0.6 ng/ml up to 27.7 ng/ml, suggesting that percentiles might be helpful in the interpretation of an individuals resistin value. While age and gender do not influence resistin levels, BMI and occurrence of diabetes have to be considered. PMID- 15523597 TI - The metabolic syndrome and incident diabetes: assessment of four suggested definitions of the metabolic syndrome in a Chinese population with high post prandial glucose. AB - AIMS: To assess the sensitivity and specificity of the four definitions of the metabolic syndrome for incident diabetes in both men and women. METHODS: The screening survey for type 2 diabetes was conducted in 1994. A follow-up study on 627 high-risk non-diabetic individuals at baseline was carried out in 1999 in Beijing area. 70 men and 76 women developed diabetes during the five-year follow up. Sensitivity and specificity of four definitions of the metabolic syndrome based on the NCEP, WHO, EGIR and AACE recommendations were compared by McNemar's test. RESULTS: The metabolic syndrome based on all four definitions identified men at a 3.7-4.5-fold and women at a 1.6-2.8-fold risk of developing diabetes during 5-year follow-up. The AACE definition had the highest sensitivity for predicting diabetes (men: 0.61; women: 0.58) and lowest specificity (men: 0.71; women: 0.70). The WHO definition identified 53 % of male and 42 % female incident diabetes. The NCEP definition of adiposity as waist girth > 102 cm was the least sensitive, detecting only 27 % of incident diabetes in men; however, it was the most specific (0.91). The EGIR definition identified the lowest number of female cases (28 %) and fewer male cases (28 %) of incident diabetes, but was specific (women: 0.87; men: 0.91). CONCLUSIONS: Further studies on definition of the metabolic syndrome should focus on the potential ethnic differences in insulin resistance and anthropometric indicators for obesity. PMID- 15523598 TI - The comparison of insulin sensitivity in non-diabetic hemodialysis patients treated with and without recombinant human erythropoietin. AB - BACKGROUND: Patients with end-stage renal disease (ESRD) are known to have insulin resistance. Treatment with EPO is associated with improvement in insulin sensitivity in uremic patients. The aim of this study was to compare insulin sensitivity and pancreatic B cell function in adult non-diabetic uremic hemodialysis patients treated with or without rHuEPO. SUBJECTS AND METHODS: Three groups of subjects were included to the study: hemodialysis patients treated with rHuEPO [EPO(+) group] or without rHuEPO [EPO(-) group], and healthy controls. Anthropometrical parameters, lipid levels, fasting glucose and insulin levels were measured in all subjects. Homeostasis Model Assessment (HOMA) was used to compare insulin sensitivity. ANOVA, independent t-test, and Pearson correlation were used for statistical analysis. RESULTS: Mean insulin level of control group (20.04 +/- 7.2 pmol/l) was significantly lower than EPO(+) group (p < 0.04) and EPO(-) group (p < 0.0001). HOMA-(%B) levels in the EPO(+) group were significantly lower than in the EPO(-) group (106 +/- 42, 140 +/- 63 respectively, p < 0.02). HOMA-(%B) levels in the control group (66 +/- 17) were significantly lower than in the EPO(+) and EPO(-) group (p < 0.005 and p < 0.0001 respectively). HOMA-(%S) levels in the EPO(+) groups was significantly higher than in the EPO(-) group (91 +/- 40, 56 +/- 26, respectively; p < 0.01). HOMA (%S) levels of control group (125 +/- 24 ) was significantly higher than EPO(+) and EPO(-) groups (p < 0.02, p < 0.0001 respectively). We found a positive correlation between duration of erythropoietin treatment and insulin sensitivity (r = 0.484, p < 0.002). CONCLUSIONS: Firstly, patients treated with EPO are insulin sensitive compared to patients not treated with EPO. Secondly, duration of erythropoietin treatment is positively correlated with insulin sensitivity in hemodialysis patients. PMID- 15523599 TI - Plasma adiponectin concentrations in patients with chronic renal failure: relationship with metabolic risk factors and ischemic heart disease. AB - AIMS: To compare plasma adiponectin levels between healthy controls and patients with chronic renal failure and to examine for a relationship between plasma adiponectin levels and ischemic heart disease as well as aortic distensibility which is an early marker of atherosclerosis. METHODS: We included 89 patients with CRF (45 on and 44 not on hemodialysis) and 70 controls in a cross-sectional study. Plasma adiponectin levels were measured by radioimmunoassay. Aortic distensibility was assessed by high-resolution ultrasonography. RESULTS: Plasma adiponectin levels were significantly almost twice as high in patients with renal failure compared to controls (9.7 +/- 1.1 vs. 5.4 +/- 0.6 microg/ml, p < 0.0001). No significant differences were found between renal patients on hemodialysis and not on hemodialysis (p = 0.71). Multivariate linear regression analysis in the renal patient group demonstrated a significant negative relationship between plasma adiponectin levels and ischemic heart disease (p = 0.02). The same analysis in the control subjects group showed a significant, negative relationship between plasma adiponectin levels and body mass index (p = 0.02) and a highly significant positive relationship with the high density lipoprotein cholesterol (p < 0.0001). In the total study population, glomerular filtration rate was the only independent predictor of plasma adiponectin concentrations. Aortic distensibility was lower in renal patients than in controls at a high level of significance (p < 0.0001). However, no significant relationship could be found between plasma adiponectin and aortic distensibility in either the controls or the renal patients. CONCLUSIONS: Plasma adiponectin levels are almost twice as high in patients with chronic renal failure in comparison with healthy controls, but not different between renal patients on and those not on hemodialysis. In addition, low plasma adiponectin levels are strongly associated with ischemic heart disease, but not with aortic distensibility in chronic renal failure. PMID- 15523600 TI - Thy-1 expression during 3T3-L1 adipogenesis. PMID- 15523601 TI - Single spin-echo proton transverse relaxometry of iron-loaded liver. AB - A single-spin-echo methodology is described for the measurement and imaging of proton transverse relaxation rates (R2) in iron-loaded and normal human liver tissue in vivo. The methodology brings together previously reported techniques dealing with (i) the changes in gain between each spin-echo acquisition, (ii) signal level offset due to background noise, (iii) estimation of signal intensities in decay curves at time zero to enable reliable extraction of relaxation times from tissues with very short T2 values, (iv) bi-exponential modelling of decay curves with a small number of data points, and (v) reduction of respiratory motion artefacts. The accuracy of the technique is tested on aqueous manganese chloride solutions yielding a relaxivity of 74.1+/-0.3 s-1 (mM) 1, consistent with previous reports. The precision of the in vivo measurement of mean liver R2 values is tested through duplicate measurements on 10 human subjects with mean liver R2 values ranging from 26 to 220 s-1. The random uncertainty on the measurement of mean liver R2 was found to be 7.7%. PMID- 15523602 TI - Vertical conjugate eye deviation in postresuscitation coma. AB - Vertical eye deviation in hypoxic coma is considered to be rare. In contrast, we found that in a consecutive series of 50 postresuscitation comatose patients, 28 (56.0%) developed tonic upward or downward eye deviation. We suggest that both the upward and the downward deviations resulted from diffuse cerebrocerebellar damage sparing the brainstem. Upward deviation is an early sign, whereas downward deviation appears later and generally implies a transition to the vegetative state. PMID- 15523603 TI - Identification of subpopulations of cells with differing telomere lengths in mouse and human cell lines by flow FISH. AB - BACKGROUND: Telomeres are specialized nucleoprotein structures at chromosome ends that undergo dynamic changes after each cell cycle. Understanding the mechanisms of telomere dynamics is critically dependent on the ability to accurately measure telomere length in a cell population of interest. Techniques such as Southern blot, which measures average telomere length, and quantitative fluorescence in situ hybridization (Q-FISH), which can estimate telomere length in individual chromosomes, are limited in their capacity to determine the distribution of cells with differing telomere lengths in a given cell population. METHODS: We employed flow-FISH to determine whether mouse and human cell lines exhibit subpopulations of cells with differing telomere lengths. RESULTS: Our analysis showed that at least one of four analyzed mouse cell lines had two subpopulations of cells with differing telomere lengths. Differences in telomere length between subpopulations of cells were significant, and we term this phenomenon TELEFLUCS (TElomere LEngth FLUctuations in Cell Subpopulations). We also observed TELEFLUCS in 1 of 19 analyzed human nonalternative lengthening of telomere cell lines and in 1 of 2 analyzed human alternative lengthening of telomere cell lines. The existence of cell subpopulations with differing telomere lengths was confirmed by Q-FISH. CONCLUSION: Our results underscore the importance of flow-FISH in telomere length analysis. PMID- 15523604 TI - Autoimmune disorders in Kabuki syndrome. AB - Kabuki syndrome is associated with abnormalities in multiple organ systems. While many of the anomalies are congenital malformations, other clinical manifestations may not appear until later in childhood. Among these associated conditions, autoimmune abnormalities have been described in several patients. These include idiopathic thrombocytopenic purpura (ITP), hemolytic anemia, thyroiditis, and vitiligo. In this report, we describe five affected patients with autoimmune manifestations. Four patients had ITP, and two of these patients had concurrent hemolytic anemia. The fifth patient had vitiligo. Two of the patients with ITP had a chronic and relapsing course. Of note, some of these patients also had hypogammaglobulinemia. The autoimmune disorders may be manifestations of abnormal immune regulation. We conclude that Kabuki syndrome is associated with an increased incidence of autoimmune disorders. In addition, the presence of an underlying immune defect may predispose these children to a chronic course of these autoimmune conditions. PMID- 15523605 TI - Intellectual abilities and adaptive behavior of children and adolescents with Kabuki syndrome: a preliminary study. AB - Very little is known about the intellectual abilities and adaptive behavior of individuals who have Kabuki syndrome, beyond the fact that most individuals with this syndrome have mental retardation. To fill this gap, we have completed psychological assessments of 11 children and adolescents with Kabuki syndrome. Results indicated that most of the participants functioned in the range of mild mental retardation, with both intellectual and adaptive behavior in the mildly deficient range and problem behaviors, if any, limited to mild difficulties with inattention and/or hyperactivity-impulsivity and mild problems with obsession/anxiety. At the lower extreme, one child evidenced severe mental retardation and profound adaptive behavior impairment accompanied by serious externalizing and internalizing problem behaviors. At the upper extreme, one adolescent had average intelligence and adaptive behavior, with problem behaviors well within the normal range for his chronological age. Most participants evidenced relative intellectual strengths in verbal and reasoning abilities and a relative weakness in visuospatial construction abilities. This pattern affected adaptive behavior as well, yielding a relative strength in Social Interaction and Communicative Skills and considerable weakness in Motor Skills and Personal Living Skills. PMID- 15523606 TI - Chromosome 22q11 deletion and pachygyria characterized by array-based comparative genomic hybridization. PMID- 15523607 TI - Pronounced short stature in a girl with tricho-rhino-phalangeal syndrome II (TRPS II, Langer-Giedion syndrome) and growth hormone deficiency. AB - We report on a 10-year-old girl with tricho-rhino-phalangeal syndrome type II (TRPS II) and pronounced short stature (-4.8 SD). The patient has an interstitial chromosome 8q24.1 deletion of 12-15 Mb. The deletion spans all genes from CSMD3 to at least ANXA13 including the TRPS1 and EXT1 genes, which are responsible for the TRPS II phenotype. In addition to the features of TRPS II, the patient had growth hormone (GH) deficiency with diminished response in three stimulation tests. Therapy with 0.2 mg GH/kg/week led to an increase of growth velocity from 2.5 to 6.6 cm/year. To our knowledge, such a combination of TRPS II and GH deficiency has not yet been described. PMID- 15523608 TI - Hippocampus and remote spatial memory in rats. AB - Damage to the hippocampus typically produces temporally graded retrograde amnesia, whereby memories acquired recently are impaired more than memories acquired remotely. This phenomenon has been demonstrated repeatedly in a variety of species and tasks. It has also figured prominently in theoretical treatments of memory and hippocampal function. Yet temporally graded retrograde amnesia has not been demonstrated following hippocampal damage in spatial tasks like the water maze. We have assessed recent and remote spatial memory following hippocampal lesions in three different tests of spatial memory: (1) the standard water maze; (2) the Oasis maze, a dry-land version of the water maze; and (3) the annular water maze, where training and testing occur within a circular corridor. Training protocols were developed for each task such that retention of spatial memory could be expressed after very long retention intervals. In addition, retention in each task was assessed with single probe trials so that the assessment of remote memory did not depend on the ability to relearn across multiple trials. The findings were consistent across the three tasks. In the standard water maze (Experiment 1), spatial memory was impaired after training surgery intervals of 1 day, 8 weeks, or 14 weeks. Similarly, in the Oasis maze (Experiment 2), spatial memory was impaired after training-surgery intervals of 1 day and 9 weeks. Finally, in the annular water maze (Experiment 3), spatial memory was impaired after training-surgery intervals of 9 weeks and 14 weeks. Dorsal hippocampal lesions impaired performance to the same extent as complete lesions. The impairment in remote spatial memory could reflect disruption of previously acquired spatial information. Alternatively, it is possible that in these tasks hippocampal lesions might produce an impairment in performance that prevents the expression of an otherwise intact spatial memory. PMID- 15523609 TI - Failure to acquire new semantic knowledge in patients with large medial temporal lobe lesions. AB - We examined new semantic learning in two profoundly amnesic patients (E.P. and G.P.) whose lesions involve virtually the entire medial temporal lobe (MTL) bilaterally. The patients were given five tests of semantic knowledge for information that could only have been acquired after the onset of their amnesia in 1992 and 1987, respectively. Age-matched and education-matched controls (n = 8) were also tested. On tests of recall, E.P. and G.P. each scored 10% correct on a test of 20 easy factual questions (controls = 90%), 2% and 4% correct on 55 questions about news events (controls = 85%), and 0% and 4% correct on a test of 24 famous faces. On three tests of recognition memory for this same material, the patients scored at chance levels. Similarly, the patients were unable to judge whether persons who had been famous for many decades were still living or had died during the past 10 years (E.P. = 53%; G.P. = 50%; controls = 73%; chance = 50%). Lastly, neither patient E.P. nor patient G.P. could draw an accurate floor plan of his current residence, despite having lived there for 10 years and 1 year, respectively. The results demonstrate that the capacity for new semantic learning can be absent, or nearly absent, when there is virtually complete damage to the MTL bilaterally. Accordingly, the results raise the possibility that the acquisition of conscious (declarative) knowledge about the world cannot be supported by structures outside the MTL, even with extended exposure. Published 2004 Wiley-Liss, Inc. PMID- 15523610 TI - Does hippocampus associate discontiguous events? Evidence from event-related fMRI. AB - To examine the hypothesis that the hippocampus is necessary to overcome temporal or spatial "discontiguity" (Wallenstein et al., Trends Neurosci 1998; 21:317 323), subjects were imaged by functional magnetic resonance imaging (fMRI) when they were making judgments as to whether two words were semantically related. Two words were presented, either at the same time (the Simultaneous Presentation Condition) or one after the other with a short unfilled rest period (the Delayed Presentation Condition). The latter condition, relative to the former, was proposed to involve the process of "discontiguity association." Event-related fMRI results of eight subjects showed that, relative to the binding of simultaneously presented words, the binding of delay presented words was associated with left hippocampus activity. This result provided direct neuroimaging evidence for the role of the hippocampus in "discontiguity association." PMID- 15523611 TI - Inhibition of hippocampal protein synthesis following recall disrupts expression of episodic-like memory in trace conditioning. AB - Transition of short-term to long-term memory is referred to as consolidation and the process is dependent on protein synthesis. Recently, several studies have shown that expression of consolidated memory for simple forms of learning tasks (e.g., delay conditioning, contextual fear, inhibitory avoidance) becomes vulnerable to disruption by inhibition of protein synthesis when administered shortly after recall. In the present study, we address whether recall-induced dependence on protein synthesis is a fundamental property that can be applied to a form of memory requiring attentional awareness or is specific to memories for simple forms of conditioning. Trace fear conditioning is a form of learning that requires an active memory trace to associate a conditioned stimulus (CS) with an unconditioned stimulus (US) separated by time. Our data demonstrate that whether a CS-alone recall trial in a novel context acts as an extinction or reactivation trial depends on the strength of the original memory. Inhibition of protein synthesis following the recall trial in animals receiving one trace conditioning training session (that gives rise to weak memory) resulted in enhanced CS elicited freezing compared with vehicle control, as a result of impaired extinction memory, but had no effect on contextual memory. However, inhibition of hippocampal protein synthesis following the recall trial in animals receiving two trace conditioning training sessions (that gives rise to stronger memory) resulted in impaired retention of both trace CS-US associative and contextual memory despite that the context-US association was not directly reactivated. This provides evidence that, for a robust memory, the CS-alone recall trial results in the reactivation of an episodic-like memory, including trace CS- and contextual memory, and that hippocampal information storage for the memory as a whole is returned to a labile state requiring de novo protein synthesis. This and other studies are consistent with the role of the hippocampus in coordinating episodic memory retrieval. PMID- 15523612 TI - Fatal lymphoproliferative disorder in a child with Schimke immuno-osseous dysplasia. AB - Schimke immuno-osseous dysplasia (SIOD) is a rare autosomal recessive disorder characterized by spondyloepiphyseal dysplasia, nephrotic syndrome, and cell mediated immunodeficiency. Mutations in the SMARCAL1 gene (SW1/SNF2-related, matrix-associated, actin-dependent regulator of chromatin, subfamily a-like1) cause SIOD. We report a patient with SIOD and SMARCAL1 mutations, who presented with fever of unknown origin secondary to B-cell lymphoma. To our knowledge, this is the first report of an SIOD patient with a primary lymphoproliferative disorder (LPD). PMID- 15523613 TI - High prevalence of isolated pericardial effusion in Down syndrome. PMID- 15523614 TI - Somatic mosaicism of chromosome 7 in a highly proliferating melanocytic congenital naevus in a ring chromosome 7 patient. AB - Ring chromosome 7 is a rare but well documented chromosomal aberration in man. So far at least 14 cases have been reported in the literature showing a variable but distinct pattern of phenotypic characteristics in affected individuals. Besides others, skin findings as pigmented naevi are especially frequent. Loss of chromosomal material from the terminal chromosome arms in the structurally abnormal ring chromosome 7 as well as somatic mosaicism with loss or gain of chromosome 7 has been suggested to be responsible for the clinical symptoms. We now report another case of a ring chromosome 7 in a 14-year-old boy with multiple remarkable congenital naevi, where we could demonstrate for the first time somatic mosaicism showing significant gain of chromosome 7 within a highly proliferating melanocytic congenital naevus (MCN). PMID- 15523615 TI - Novel mutation in the tyrosine kinase domain of FGFR2 in a patient with Pfeiffer syndrome. AB - Mutations in the fibroblast growth factor receptor 2 (FGFR2) cause a variety of craniosynostosis syndromes. The mutational spectrum tends to be narrow with the majority of mutations occurring in either exon IIIa or IIIc or in the intronic sequence preceding exon IIIc. Mutations outside of this hotspot are uncommon and the few identified mutations have demonstrated wide clinical variability, making it difficult to establish a clear-cut genotype-phenotype correlation. To better delineate the clinical picture associated with these unusual mutations, we describe a severely affected patient with Pfeiffer syndrome and a missense mutation in the tyrosine kinase (TK) domain of FGFR2. PMID- 15523617 TI - Subcutaneous neurofibromas are associated with mortality in neurofibromatosis 1: a cohort study of 703 patients. AB - Neurofibromatosis 1 (NF1) is a common genetic disorder with an autosomal dominant mode of inheritance, an increased morbidity and mortality, and a shorter lifespan. Although the disease is fully penetrant by the age of 8, the variability in symptoms and complications is high, even among members of the same family. The aim of this study was to identify easily recognizable clinical features that may be associated with mortality in a cohort of patients affected with NF1. We used prospectively collected data from the Neurofibromatosis Institute Database (NFID) and included in our analysis 703 patients who fulfilled the NIH diagnostic criteria for NF1. Clinical, especially dermatological features were tested as potential factors associated with mortality. Among the patients, 405 (57.6%) were children and 298 (42.4%) were adults. The mean follow-up was 2.4 years (median = 0.98, range: 0-15.3 years). Forty patients died during follow-up, mostly due to tumor development such as sarcoma (n = 18). In the adult population, subcutaneous neurofibromas (odds ratio [OR] = 3.6, 95% confidence interval (CI): [1.2-11.3], P = 0.02) and male gender (OR = 5.6, [1.5-20.9], P = 0.004) were independent predictors of mortality after adjustment for age. Among children, the presence of facial plexiform neurofibromas and pruritus were significantly associated with mortality in univariate analysis. Our study describes independent risk factors of mortality in a large cohort of adult and pediatric patients. Close follow-up should be obtained for patients presenting with subcutaneous neurofibromas. PMID- 15523618 TI - Two sisters with Silver-Russell phenotype. AB - Silver-Russell syndrome (SRS) is a well recognizable syndrome, but the etiology of SRS seems to be heterogeneous. SRS is listed in Mendelian Inheritance in Man as an autosomal dominant disorder because most described cases have been of sporadic occurrence, and most likely were caused by de novo autosomal dominant mutation, and because families with apparent dominant transmission of a SRS phenotype have been described. Still, in a few families, autosomal recessive inheritance has been suggested. We describe two sisters who meet the criteria for SRS proposed by Price et al. [1999]. The parents had normal facial features, normal height, and normal post-natal growth. This is the second well-documented case of familial recurrence of SRS that resembles an autosomal recessive inheritance pattern. Since sib recurrence is so rare in SRS, other modes of inheritance should be considered. The finding of maternal uniparental disomy 7 (mUPD7) in 10% of SRS cases suggests that lack of paternally expressed imprinted gene(s) or overexpression of maternal imprinted gene(s) on chromosome 7 cause SRS. The recurrence in sibs could be caused by a mutation in the imprinted gene or imprinting center carried by one parent. Alternatively, recurrence in sibs could represent germ line mosaicism for a dominant mutation in one of the parents. PMID- 15523619 TI - Novel and recurrent exon 13 mutations of COMP in pseudoachondroplasia. PMID- 15523620 TI - KBG syndrome in a cohort of Italian patients. AB - KBG syndrome comprises a distinct facial phenotype, macrodontia, short stature, and skeletal anomalies. So far, it has been reported in 29 individuals. Recently, diagnostic criteria were outlined. Here, we describe eight new patients whose clinical and radiological findings fit the diagnostic criteria of KBG syndrome. While most patients were sporadic in occurrence, in two families the disorder was transmitted from mildly affected mothers to their affected children. The phenotype of KBG syndrome has been reviewed based on published and present patients. EEG anomalies with or without seizures, mixed hearing loss, palatal anomalies with secondary speech disorder, distinct age-related behavior, and cryptorchidism are possible additional characteristics. Less common manisfestations were posterior fossa malformations, eye defects, and congenital heart defects. PMID- 15523621 TI - Histological structure of antlers in castrated male fallow deer (Dama dama). AB - Antlers are periodically replaced cranial appendages that, except for the reindeer, are grown only by male deer. The annual antler cycle is controlled by seasonal fluctuations of sex steroid concentrations in the blood, and accordingly castration of male deer causes deviations from normal antler growth. The present study investigated antler histology of castrated fallow bucks (Dama dama). Castration in early spring was followed by casting of the hard antlers carried by the bucks and the growth of a new set of antlers, which remained in velvet permanently. In the following year, numerous bony protuberances developed from the original antler surface. Further growth of these protuberances, which were formed by subperiosteal intramembranous ossification, led to a marked increase in antler diameter in the affected areas. Compared to antlers of intact bucks, the antlers of the castrates showed histological signs of immaturity, suggestive of a reduced bone remodeling and an impairment of the mineralization process. These changes point to the dependence of the above processes on a stimulation by higher levels of sex steroids. Two years after castration, the antlers also developed integumental thickening and showed an initial formation of skin outgrowths. Cystic structures were present in the skin, which were often filled with a presumably sebaceous and/or keratinous material. Formation of intradermal bone or cartilage was not observed in the antlers of the castrated fallow bucks. The histological structure of the skin outgrowths suggested that they were caused by a hypertrophy of the dermal component of the velvet. Due to the localized bone overgrowth, resulting from the periosteal bone apposition onto the original antler surface, skin-lined infoldings originated, which reached deep into the newly formed bone. Our study revealed no indication of invasive/destructive bone growth in the antlers, i.e., of a penetration of the newly formed bone tissue into the pre-existing bone. The hypertrophic bone growth in the antlers of the castrates is compared with other forms of periosteally derived hypertrophic bone formation, including osteomas, in the mammalian skeleton. It is discussed whether the skin and bone outgrowths of the antlers of castrated fallow bucks may be classified as benign tumors. PMID- 15523622 TI - Genotype-phenotype correlations for a wide spectrum of mutations in the Wilson disease gene (ATP7B). AB - Wilson disease (WND) is caused by mutations in the ATP7B gene and exhibits substantial allelic heterogeneity. In this study we report the results of molecular analyses of 20 WND families not described previously. When combined with our prior results, the cohort includes 93 index patients from 69 unrelated families. Twenty different mutations accounted for 86% of the WND chromosomes. The most frequent were p.H1069Q (35%), p.R969Q (12%), c.2530delA (7%), p.L936X (7%), p.Q289X (7%), and p.I1148T (3%). We also present here a detailed phenotypic assessment for patients whose molecular result was previously reported. Thirty cases were homozygous for 9 different mutations, 13 of which were homozygous for p.H1069Q, and 7 for p.R969Q. Mutations p.H1069Q and p.R969Q appeared to confer a milder disease as patients showed disease onset at a later age, and were associated with milder severity when found in trans with severe mutations. Predicted nonsense and frameshift mutations were associated with severe phenotypic expression with earlier disease onset and lower ceruloplasmin values. WND can be treated by copper-chelation therapy, particularly if the disease is diagnosed before irreversible tissue damage occurs. Our results on the effect of predicted nonsense and frameshift mutations are especially important for early medical intervention in presymptomatic infants and children with these genotypes. PMID- 15523623 TI - Symptomatic Chiari I malformation in Kabuki syndrome. AB - Kabuki (Niikawa-Kuroki) syndrome is associated with a characteristic facial appearance, cleft palate, congenital heart defects, and developmental delay. Structural brain anomalies have only occasionally been described in Kabuki syndrome. Chiari type I malformation, characterized by caudal herniation of the cerebellar tonsils through the foramen magnum, has been described only infrequently in association with defined syndromes and has been reported once in association with Kabuki syndrome. We report three additional children with Kabuki syndrome who have Chiari I malformation. Two children presented with chronic headaches and the third patient presented with gait abnormalities in adolescence. The incidence of Chiari I malformation may be higher than previously reported in Kabuki syndrome since it may not be diagnosed until later in childhood, whereas most reports of Kabuki syndrome are of young children. Further, symptoms of Chiari I anomaly can be somewhat nonspecific. Thus, we suggest that Chiari type I be considered in patients with Kabuki syndrome who present with persistent headache, neck pain, or other symptoms suggestive of Chiari I anomaly. PMID- 15523624 TI - Phenotypic and molecular characterization of Bruck syndrome (osteogenesis imperfecta with contractures of the large joints) caused by a recessive mutation in PLOD2. AB - Bruck syndrome (BS) is a recessively-inherited phenotypic disorder featuring the unusual combination of skeletal changes resembling osteogenesis imperfecta (OI) with congenital contractures of the large joints. Clinical heterogeneity is apparent in cases reported thus far. While the genes coding for collagen 1 chains are unaffected in BS, there is biochemical evidence for a defect in the hydroxylation of lysine residues in collagen 1 telopeptides. One BS locus has been mapped at 17p12, but more recently, two mutations in the lysyl hydroxylase 2 gene (PLOD2, 3q23-q24) have been identified in BS, showing genetic heterogeneity. The proportion of BS cases linked to 17p22 (BS type 1) or caused by mutations in PLOD2 (BS type 2) is still uncertain, and phenotypic correlations are lacking. We report on a boy who had congenital contractures with pterygia at birth and severe OI-like osteopenia and multiple fractures. His urine contained high amounts of hydroxyproline but low amounts of collagen crosslinks degradation products; and he was shown to be homozygous for a novel mutation leading to an Arg598His substitution in PLOD2. The mutation is adjacent to the two mutations previously reported (Gly601Val and Thr608Ile), suggesting a functionally important hotspot in PLOD2. The combination of pterygia with bone fragility, as illustrated by this case, is difficult to explain; it suggests that telopeptide lysyl hydroxylation must be involved in prenatal joint formation and morphogenesis. Collagen degradation products in urine and mutation analysis of PLOD2 may be used to diagnose BS and differentiate it from OI. PMID- 15523625 TI - Heterogeneous basis of the type VIB form of Ehlers-Danlos syndrome (EDS VIB) that is unrelated to decreased collagen lysyl hydroxylation. AB - Skin fibroblasts from the majority of patients with the clinical diagnosis of Ehlers-Danlos syndrome type VI (EDS VI; kyphoscoliosis type), have significantly decreased lysyl hydroxylase (LH) activity due to mutations in the LH1 gene (classified as EDS VIA: OMIM no. 225400). A rare condition exists in which patients are clinically similar but have normal levels of LH activity (designated EDS VIB: OMIM no. 229200). To define the biochemical defect, we have examined cultured fibroblasts from four EDS VIB patients for changes in the levels of the mRNAs for LH1, LH2, and LH3, collagen cross-linking patterns, and the extent of lysine hydroxylation of type I collagen alpha chains. Although normal levels of LH1 mRNA were observed in all four patients, in two patients the levels of LH2 mRNA were decreased by >50%, and a similar decrease was observed in LH3 mRNA in the other two patients. A distinct pattern of collagen cross-links, indicative of decreased lysyl hydroxylation, could be identified in EDS VIA patients, but there was no clear correlation between collagen cross-link pattern and changes in the individual LH mRNAs in EDS VIB patients. Linkage to tenascin-X was excluded in these patients. This study suggests that the basis for this form of EDS VI is genetically heterogeneous, and that alternative pathways in addition to lysine hydroxylation of collagen may be affected. PMID- 15523626 TI - Functional analysis of polymorphic variation within the promoter and 5' untranslated region of the neurofibromatosis type 1 (NF1) gene. AB - The regulatory regions of the neurofibromatosis type 1 (NF1) gene have scarcely been screened either for mutations of potential pathological importance or for functional polymorphisms. To address this question, a 987 bp sequence spanning the promoter and 5' flanking sequence of the human NF1 gene was screened for sequence variants in 570 unrelated NF1 patients and 105 controls. Five novel sequence variants were identified, comprising a 14 bp deletion at -142 within the promoter region, three single nucleotide substitutions in the 5'UTR (C + 247T, C + 261G, G + 462C), and a substitution (C + 514T) at the 5' end of the coding region that served to generate a Stop codon. The latter is likely to be of pathological significance since it is predicted to lead to the synthesis of a truncated protein. The functional significance of three of the other variants (14 bp del, C + 261G, G + 462C) was explored by luciferase reporter gene expression and electrophoretic mobility shift assays. The del14 variant demonstrated allele specific protein binding without altered reporter gene expression and the G + 462C allele showed slightly decreased reporter gene expression. PMID- 15523627 TI - Association of partial trisomy 9p and the Dandy-Walker malformation. PMID- 15523628 TI - A founder effect in three large Newfoundland families with a novel clinically variable spastic ataxia and supranuclear gaze palsy. AB - A distinctive slowly progressive neurodegenerative disorder, which falls under a new category of neurological diseases, the hereditary spastic ataxias (HSA), is described in three independently ascertained Newfoundland kindreds. HSA is a heterogeneous group of disorders in which pyramidal tract features overlap cerebellar characteristics. The families are assumed to have the same condition as, although apparently unrelated, all originate in a historically isolated cluster of rural communities and link to the same locus at 12p13, SAX1. Clinically the phenotype is very variable but lower limb hypertonicity and hyperreflexia are early and prominent generally preceded by eye movement abnormality, an impaired vertical downward saccade and a typical involuntary head jerk. These are followed by variable levels of ataxia, dysarthria, and dysphagia. Onset occurs in the first two decades and can be detected in most by early adulthood. Significant mobility problems are present by the fourth decade with a broad based ataxic and spastic gait. MRI scans of brain and spinal cord were normal. Neuropathology showed degeneration of corticospinal tracts and posterior columns and midbrain neuronal loss. The phenotype is striking in its diversity among and within families and the variability of expression can be observed within the same sibship. Pedigree analysis shows no evidence of anticipation or any sex differences in severity. The condition is unusually prevalent in the province of Newfoundland, which is characteristic of a founder effect followed by isolation and large family size. Fine mapping efforts have reduced the critical interval of the SAX1 locus to 1.9Mb. Identification of the SAX1 gene will help to clarify the pathogenesis of this type of HSA. PMID- 15523630 TI - Cumming syndrome with heterotaxia, campomelia and absent uterus/fallopian tubes. PMID- 15523631 TI - Unexpected death due to refractory metabolic acidosis and massive hemolysis in a young infant with Prader-Willi syndrome. PMID- 15523632 TI - Autosomal dominant flat umbilicus. AB - We describe a Chinese family among whom five members in three generations had a flat umbilicus with vertical and male to male transmission indicative of autosomal dominant inheritance. Except the proband, who also had Tetralogy of Fallot, other affected members had a flat umbilicus as an isolated anomaly. Autosomal dominant transmission of isolated flat umbilicus has not been reported previously. PMID- 15523633 TI - A new three-generational family with frontometaphyseal dysplasia, male-to-female transmission, and a previously reported FLNA mutation. PMID- 15523634 TI - Differential expression of Pax6 and Ngn2 between pair-generated cortical neurons. AB - Progenitor cells that generate neuron pairs ("pair progenitor cells") are implicated in mammalian cortical development, and their division has been thought to be "symmetric." However, asymmetric growth of two sister neurons generated by the division of a pair progenitor cell would lead to more efficient generation of neuronal diversity in the cortex. To explore mechanisms by which pair progenitor cells provide neuronal diversity, we examined molecular differences between a pair of neurons generated in clonal-density culture. Time-course analysis for the acquisition of neuronal markers and the disappearance of Pax6 and Neurogenin2 (Ngn2) demonstrated that 1) these transcription factors are expressed transiently in some but not all young neurons and 2) some neuron pairs showed uneven/asymmetric expression of Pax6 (19.5%) or Ngn2 (23.8%), whereas other pairs were either symmetrically positive or negative. Asymmetric Pax6 distribution in neuron pairs was not associated with asymmetric distribution of Numb, which raises an intriguing possibility, that Pax6 asymmetry in neuron pairs is produced by an alternative mode of the cell autonomous mechanisms. Stage-dependent changes were noted in the pattern of Ngn2 retention in daughter neurons, reflecting qualitative changes in the pair progenitor population. We suggest that pair progenitor cells contribute to the generation of neuronal diversity through cell intrinsic heterogeneity and asymmetric division. PMID- 15523635 TI - 3alpha,5alpha-Tetrahydroprogesterone (allopregnanolone) and gamma-aminobutyric acid: autocrine/paracrine interactions in the control of neonatal PSA-NCAM+ progenitor proliferation. AB - The earliest identified neonatal neural progenitors are cells that express the polysialylated form of the neural cell adhesion molecule (PSA-NCAM). One of these progenitors is the early PSA-NCAM+ progenitor (ePSA-NCAM+ progenitor; Gago et al. [2003] Mol Cell Neurosci 22:162-178), which corresponds to a multipotential cell with a default differentiation through glial lineages. The ePSA-NCAM+ progenitor can synthesize the neurosteroid progesterone (PROG) and its reduced metabolite 3alpha,5alpha-tetrahydroprogesterone (3alpha,5alpha-THP, or allopregnanolone; Gago et al. [ 2001] Glia 36:295-308). The latter is a potent positive allosteric modulator of gamma-aminobutyric acid type A (GABAA) receptors. In the present work, we demonstrate that PROG and 3alpha,5alpha-THP both stimulate ePSA-NCAM+ progenitor proliferation. PROG exerted its mitogenic effect indirectly, through its conversion to 3alpha,5alpha-THP, since it could be abolished by an inhibitor of the 5alpha-reductase (L685-273) and mimicked by 3alpha,5alpha-THP. A dose response curve revealed a bell-shaped effect of 3alpha,5alpha-THP on ePSA-NCAM+ progenitor proliferation, with greatest stimulation at nanomolar concentrations. The mitogenic effect of 3 alpha,5 alpha-THP was mediated by GABAA receptors, insofar as it could be blocked by the selective antagonist bicuculline. ePSA NCAM+ progenitors indeed expressed mRNAs for GABAA receptor subunits, and GABA enhanced cell proliferation, an effect that was also bicuculline sensitive. Moreover, these cells synthesized GABA, which was involved in a tonic stimulation of their proliferation. These results reveal complex autocrine/paracrine loops in the control of ePSA-NCAM+ progenitor proliferation, involving both neurosteroid and GABA signaling, and suggest a novel key role for 3alpha,5alpha-THP in the development of the nervous system. PMID- 15523636 TI - Developmental outcome in Kabuki syndrome. AB - Over the last 20 years, a wide spectrum of congenital anomalies have been described in association with Kabuki syndrome (KS). However, very little information is available on developmental outcome. As more individuals with this syndrome are recognized and reported, it appears that as many as one-sixth may have normal intelligence. The purpose of this report is to describe the developmental outcome in 15 patients with KS, to determine whether a recognizable pattern of disabilities exist, and whether developmental outcome correlates with the presence of malformations. We ascertained 15 patients with KS from three dysmorphology and clinical genetics services in which developmental milestones and formal developmental testing were available. Based on these patients and a review of the literature, in the absence of major structural brain anomalies, the average intelligence quotient (IQ) in patients with this condition fall within the mild mental retardation range, however, specific developmental outcomes are widely variable, ranging from severe MR to normal intelligence. The presence or absence of hearing loss or major malformations, other than those involving the brain, was not predictive of developmental outcome. PMID- 15523637 TI - Neurotoxic prostaglandin J2 enhances cyclooxygenase-2 expression in neuronal cells through the p38MAPK pathway: a death wish? AB - The role of the proinflammatory and inducible form of cyclooxygenases (COX-2) in neurodegeneration is not well defined. Some of its metabolic products, such as prostaglandins (PG) of the J2 series, are known to be neurotoxic. Here we demonstrate that PGJ2 enhances COX-2 gene expression without elevating COX-1 levels in neuronal cells. PGJ2 also increased PGE2 production, establishing that the de novo synthesized COX-2 is enzymatically active. PGJ2 derivatives, such as 15d-PGJ2, are known activators of PPARgamma, a nuclear receptor that activates gene expression. However, the selective PPARgamma agonist ciglitazone failed to up-regulate COX-2, indicating that the PGJ2 effect on COX-2 is PPARgamma independent. Furthermore, PGJ2 stabilized IkappaBalpha levels, indicating that NFkappaB is not active under these conditions. The blocking of neuronal NFkappaB activity by PGJ2 may be an important contributor to its neurotoxicity, insofar as NFkappaB transactivation seems to be required for neuronal survival in the CNS. Interleukin-1 (IL1) is a proinflammatory cytokine known to stimulate the expression of genes associated with inflammation, including COX-2. Notably, IL1 mRNA levels in the neuronal cells were increased by PGJ2 treatment. The proinflammatory cytokine may mediate COX-2 up-regulation by PGJ2 through p38MAPK and not JNK activation, in that only an inhibitor of the former prevented the COX 2 increase. Thiol-reducing agents, such as N-acetylcysteine, protected the neuronal cells from the deleterious effects of PGJ2, whereas ascorbic acid did not. Collectively, our findings suggest that proinflammatory conditions that lead to COX-2 up-regulation and the concomitant production of PGJ2 initiate a mechanism of self-destruction through an autotoxic loop between PGJ2 and COX-2 that may exacerbate neurodegeneration beyond a point of no return. Thiol-reducing antioxidants may offer an optimal strategy for halting this neurodegenerative process. PMID- 15523638 TI - Craniofacial structure in Marfan syndrome: a cephalometric study. AB - Marfan syndrome (MFS) is a connective tissue disorder with autosomal dominant inheritance. Mutations in the FBN1 gene cause deficient processing of fibrillin 1, the main constituent of extracellular microfibrils, affecting tissues displaying elastic properties. Clinical manifestations are widespread and involve the skeletal, ocular, cardiovascular and pulmonary systems, skin and integumentum, and dura. A highly arched palate and retrognathia have been assigned to the symptoms with minor diagnostic specificity, although epidemiological data on prevalence are lacking yet. Twenty-six patients with MFS (n = 26) were studied for craniofacial characteristics using cephalometric measurements on lateral cranial radiographs. The purposes of this study were (1) to compare cephalometric variables of MFS group with age- and sex-matched population norms, and (2) to assess differences in palatal vault dimensions among adult MFS (n = 17) and matched controls (n = 32) by means of cephalometric measurements. Significant differences with population norms were found in the structures of the cranial base, the maxillary complex, the mandible body, and the relations of the jaws with respect to the cranial base and to each other. Palatal height and palatal length were significantly larger in MFS, and were significantly correlated to each other and to the height of the maxillo-alveolar processus. The present data disprove in part previously reported findings, possibly due to biased patient selection in these studies or demographic differences. However, a strong correlation was found between maxillary/mandibular retrognathia, long face, highly arched palate, and MFS. A combination of both intrinsic genetic factors and environmental factors is suggested as a possible explanation for specific morphogenetic aspects of the craniofacial complex in MFS. PMID- 15523639 TI - A novel frameshift mutation of FOXC2 gene in a family with hereditary lymphedema distichiasis syndrome associated with renal disease and diabetes mellitus. AB - Lymphedema-distichiasis (LD) syndrome is a clinically variable autosomal dominant disorder. The disorder is caused by mutations in the forkhead transcription factor FOXC2 gene on chromosome band 16q24.3. Here, we report the sequence of the FOXC2 gene in a German-Irish family with LD in six affected relatives over three generations and identify a single adenine base pair insertion at nt 1006--1007. This insertion creates a frameshift mutation that predicts a premature stop at codon 462. In addition to LD, four of the affected family members have renal disease and three have diabetes mellitus (DM), not usually seen in the LD syndrome. Polymorphisms of FOXC2 in diabetics have been studied in different populations. Our sequence analysis of the 5' untranslated region (UTR) C-512T shows the homozygous T allele in all family members tested. The sequencing data in this family suggests the possibility of a novel phenotype-haplotype. This novel phenotype, LD/renal disease/type 2 diabetes, might be the result of a combination of the nt 1006--1007 insA and the upstream UTR homozygous T polymorphism. PMID- 15523640 TI - A dysmorphic boy with 4qter deletion and 4q32.3-34.3 duplication: clinical, cytogenetic, and molecular findings. AB - An infant boy presented with trigonocephaly, mild craniofacial features, a small VSD, open ductus Botalli (ODB), bilateral hip dysplasia, psychomotor retardation, and hypotonia. The karyotype was 46,XY,del(4)(q34). Unexpectedly, fluorescence in situ hybridization (FISH) studies revealed not only a deletion but also a duplication. The deletion extends from 4qter to 4q34.3 and the duplication from 4q32.3 to q34.3. This is the first description of a deletion inverted duplication 4q. Possible mechanisms we can envision by which this deletion/duplication arose could be a U-type exchange causing end-to-end fusion or a two step event with a paracentric inversion and subsequent cross-over in the inverted segment. This observation suggests that the karyotype of patients with a 4q deletion should be confirmed by molecular cytogenetics. PMID- 15523642 TI - Disruption of exonic splicing enhancer elements is the principal cause of exon skipping associated with seven nonsense or missense alleles of NF1. AB - Nonsense, missense, and even silent mutation-associated exon skipping is recognized in an increasing number of genes as a novel form of splicing mutation. The analysis of individual mutations of this kind can shed light on basic pre mRNA splicing mechanisms. Using cDNA-based mutation detection analysis, we have identified one missense and six nonsense mutations that lead to different extents of exon-lacking transcripts in neurofibromatosis type 1 (NF1) patients. We confirmed mutation-associated exon skipping in a heterologous hybrid minigene context. There is evidence that the disruption of functional exonic splicing enhancer (ESE) sequences is frequently the mechanism underlying mutation associated exon skipping. Therefore, we examined the wild-type and mutant NF1 sequences with two available ESE-prediction programs. Either or both programs predicted the disruption of ESE motifs in six out of the seven analyzed mutations. To ascertain the function of the predicted ESEs, we quantitatively measured their ability to rescue splicing of an enhancer-dependent exon, and found that all seven mutant ESEs had reduced splicing enhancement activity compared to the wild-type sequences. Our results suggest that the wild-type sequences function as ESE elements, whose disruption is responsible for the mutation-associated exon skipping observed in the NF1 patients. Further, this study illustrates the utility of ESE-prediction programs for delineating candidate sequences that may serve as ESE elements. However, until more refined prediction algorithms have been developed, experimental data, preferably from patient tissues, remain indispensable to assess the clinical significance, particularly of missense and silent mutations, and to understand the structure function relationship in the corresponding protein. PMID- 15523643 TI - Sox10 acts as a tissue-specific transcription factor enhancing activation of the myelin basic protein gene promoter by p27Kip1 and Sp1. AB - Myelin basic protein (MBP) is one of the major components of the myelin sheath that insulates axons. In the central nervous system, MBP is synthesized by differentiating oligodendrocytes. The expression of MBP in oligodendrocytes is regulated mainly at the transcriptional level. The Sp1 family of transcription factors has been shown to be important in the regulation of many genes. Binding of Sp1 to the GC box in the proximal MBP promoter has been shown to be indispensable for the activation of MBP gene expression. Previous results from our laboratory have shown that the increase in p27Kip1 that accompanies oligodendrocyte differentiation is paralleled by an increase in Sp1. We also have shown that the increase in MBP expression resulting from elevated p27Kip1 levels is mediated through Sp1 and that this effect occurs specifically in oligodendrocytes. In this study, we found that increased expression of p27Kip1 together with the nervous-system-specific transcription factor Sox10 can activate the MBP promoter even in nonoligodendrocyte cells. This indicates that Sox10 confers cell type specificity on the expression of MBP. Both Sp1 and Sox10 can enhance MBP promoter activity when expressed alone. Cotransfection of plasmids encoding Sp1 and Sox10 induces increased activation of the MBP promoter over expression of either transcription factor alone. This effect is not limited to oligodendrocyte cell lines, in that Sp1 and Sox10 can also synergistically activate the MBP promoter when expressed in NIH3T3 fibroblasts. Mutation of the Sp1 binding sites in the MBP promoter eliminates Sox10 stimulated activation, suggesting that the MBP promoter is activated, at least in part, through protein protein interactions between Sp1 and Sox10. PMID- 15523644 TI - A common nonsense mutation in the repetitive Kringle IV-2 domain of human apolipoprotein(a) results in a truncated protein and low plasma Lp(a). AB - LPA, the gene coding for apolipoprotein(a) [apo(a)], is the major determinant of lipoprotein(a) [Lp(a)] plasma levels, which are associated with risk for coronary heart disease (CHD) and stroke. It is not completely understood how variation in LPA relates to Lp(a) concentrations. One type of variation related to Lp(a) levels is the number of Kringle (K) IV-2 (g.61C>T; GenBank L14005.1) repeats in LPA, but sequence variation may also contribute. Human apo(a) contains from two to >40 nearly identical K IV-2 repeats of genomic size 5.5 kb, which makes it difficult to detect mutations. To elucidate the genetic variation of the apo(a) K IV-2 domain, we isolated a single "nonexpressing" apo(a) allele with 26 K IV-2 repeats, followed by PCR, cloning and sequencing of 96 clones, resulting in an average coverage of each K IV-2 repeat of approximately four-fold. The previously described K IV types 2A and 2B (K IV-2A and K IV-2B) were detected in 74% of the clones. In addition, a new type designated 2C (K IV-2C) was present. A nonsense mutation in the first exon of K IV-2 (g.61C>T) predicted to result in a truncated protein (p.R21X) was found in nine clones on a K IV-2A background. The presence of this mutation was confirmed by analysis of genomic DNA and was shown to represent the rare allele (frequency 0.02) of a SNP. Immunoblot analysis of apo(a) from plasma confirmed the presence of a truncated apo(a) isoform in the index individual and family members. Our data show that SNPs affecting Lp(a) plasma concentrations also exist in the apo(a) K IV-2 domain. PMID- 15523645 TI - Identification and characterization of novel mutations of the major Fanconi anemia gene FANCA in the Japanese population. AB - Fanconi anemia (FA) is a rare autosomal recessive disorder of hematopoiesis, with at least 11 complementation groups. FANCA, a gene for group A, accounts for the majority of FA patients. Previous studies of FANCA mutations revealed high allelic heterogeneity, frequent occurrence of large deletions, and interpopulation differences. However, systematic mutational analysis, including gene dosage assay to detect large deletions, has not been documented for Asian populations. A newly developed TaqMan quantitative PCR-based gene dosage assay, combined with sequencing of exons and cDNA fragments, allowed for detection of 48 mutant alleles of FANCA in 27 (77%) of 35 unrelated Japanese FA families with no detectable mutations in FANCC or FANCG. We identified 29 different mutations (21 nucleotide substitutions or small deletions/insertions and eight large deletions), at least 20 of which were novel. The FANCA mutational spectrum of the Japanese was different from that of other ethnic groups so far studied. This is the largest scale of mutation analysis of FANCA in the Japanese population. Characterization of these mutations provided new information regarding the mutagenesis mechanisms and structure-function relationship of FANCA. Specifically, our data suggest that diverse mechanisms including nonhomologous recombination as well as Alu-mediated homologous recombination are involved in the generation of large deletions in FANCA. PMID- 15523646 TI - LDL-receptor mutations in Europe. AB - Familial hypercholesterolemia (FH) is a clinical definition for a remarkable increase of cholesterol serum concentration, presence of xanthomas, and an autosomal dominant trait of either increased serum cholesterol or premature coronary artery disease (CAD). The identification of the low-density lipoprotein (LDL)-receptor (LDLR) as the underlying cause and its genetic characterization in FH patients revealed more insights in the trafficking of LDL, which primarily transports cholesterol to hepatic and peripheral cells. Mutations within LDLR result in hypercholesterolemia and, subsequently, cholesterol deposition in humans to a variable degree. This confirms the pathogenetic role of LDLR and also highlights the existence of additional factors in determining the phenotype. Autosomal dominant FH is caused by LDLR deficiency and defective apolipoprotein B 100 (APOB), respectively. Heterozygosity of the LDLR is relatively common (1:500). Clinical diagnosis is highly important and genetic diagnosis may be helpful, since treatment is usually effective for this otherwise fatal disease. Very recently, mutations in PCSK9 have been also shown to cause autosomal dominant hypercholesterolemia. For autosomal recessive hypercholesterolemia, mutations within the so-called ARH gene encoding a cellular adaptor protein required for LDL transport have been identified. These insights emphasize the crucial importance of LDL metabolism intra- and extracellularly in determining LDL-cholesterol serum concentration. Herein, we focus on the published European LDLR mutation data that reflect its heterogeneity and phenotypic penetrance. PMID- 15523647 TI - Enchondromatosis (Ollier disease, Maffucci syndrome) is not caused by the PTHR1 mutation p.R150C. AB - Enchondromatosis (Ollier disease, Maffucci syndrome) is a rare developmental disorder characterized by multiple enchondromas. Not much is known about its molecular genetic background. Recently, an activating mutation in the parathyroid hormone receptor type 1 (PTHR1) gene, c.448C>T (p.R150C), was reported in two of six patients with enchondromatosis. The mutation is thought to result in upregulation of the IHH/PTHrP pathway. This is in contrast to previous studies, showing downregulation of this pathway in other cartilaginous tumors. Therefore, we investigated PTHR1 in enchondromas and chondrosarcomas from 31 enchondromatosis patients from three different European countries, thereby excluding a population bias. PTHR1 protein expression was studied using immunohistochemistry, revealing normal expression. The presence of the described PTHR1 mutation was analyzed, using allele-specific oligonucleotide hybridization confirmed by sequence analysis, in tumors from 26 patients. In addition, 11 patients were screened for other mutations in the PTHR1 gene by sequence analysis. Using both allele-specific oligonucleotide hybridization and sequencing, we could neither confirm the previously found mutation nor find any other mutations in the PTHR1 gene. These results indicate that the PTHR1 gene is not, in contrast to previous suggestions, the culprit for enchondromatosis. PMID- 15523648 TI - Transfer of a mitochondrial DNA fragment to MCOLN1 causes an inherited case of mucolipidosis IV. AB - A patient with mucolipidosis-IV heterozygous for two mutations in MCOLN1 expressed only her father's cDNA mutation c.1207C>T predicting an R403C change in mucolipin. She inherited a 93bp segment from mitochondrial NADH dehydrogenase 5 (MTND5) from her mother that was inserted in-frame prior to the last nucleotide of exon 2 of MCOLN1 (c.236_237ins93). This alteration abolished proper splicing of MCOLN1. The splice site at the end of the exon was not used due to an inhibitory effect of the inserted segment, resulting in two aberrant splice products containing stop codons in the downstream intron. These products were eliminated via nonsense-mediated decay. This is the first report of an inherited transfer of mitochondrial nuclear DNA causing a genetic disease. The elimination of the splice site by the mitochondrial DNA requires a change in splicing prediction models. PMID- 15523649 TI - High-Density SNP genotyping defines 17 distinct haplotypes of the TNF block in the Caucasian population: implications for haplotype tagging. AB - The region spanning the tumor necrosis factor (TNF) cluster in the human major histocompatibility complex (MHC) has been implicated in susceptibility to numerous immunopathological diseases, including type 1 diabetes mellitus and rheumatoid arthritis. However, strong linkage disequilibrium across the MHC has hampered the identification of the precise genes involved. In addition, the observation of "blocks" of DNA in the MHC within which recombination is very rare, limits the resolution that may be obtained by genotyping individual SNPs. Hence a greater understanding of the haplotypes of the block spanning the TNF cluster is necessary. To this end, we genotyped 32 human leukocyte antigen (HLA) homozygous workshop cell lines and 300 healthy control samples for 19 coding and promoter region SNPs spanning 45 kb in the central MHC near the TNF genes. The workshop cell lines defined 11 SNP haplotypes that account for approximately 80% of the haplotypes observed in the 300 control individuals. Using the control individuals, we defined a further six haplotypes that account for an additional 10% of donors. We show that the 17 haplotypes of the "TNF block" can be identified using 15 SNPs. PMID- 15523650 TI - Nonsense-mediated and nonstop decay of ribosomal protein S19 mRNA in Diamond Blackfan anemia. AB - Mutations in the ribosomal protein (RP)S19 gene have been found in about 25% of the cases of Diamond-Blackfan anemia (DBA), a rare congenital hypoplastic anemia that includes variable physical malformations. Various mutations have been identified in the RPS19 gene, but no investigations regarding the effect of these alterations on RPS19 mRNA levels have been performed. It is well established that mutated mRNA containing a premature stop codon (PTC) or lacking a stop codon can be rapidly degraded by specific mechanisms called nonsense mediated decay (NMD) and nonstop decay. To study the involvement of such mechanisms in DBA, we analyzed immortalized lymphoblastoid cells and primary fibroblasts from patients presenting different kinds of mutations in the RPS19 gene, generating allelic deletion, missense, nonsense, and nonstop messengers. We found that RPS19 mRNA levels are decreased in the cells with allelic deletion and, to a variable extent, also in all the cell lines with PTC or nonstop mutations. Further analysis showed that translation inhibition causes a stabilization of the mutated RPS19 mRNA. Our findings indicate that NMD and nonstop decay affect the expression of mutated RPS19 genes; this may help to clarify genotype-phenotype correlations in DBA. PMID- 15523651 TI - Functional characterization of SIX3 homeodomain mutations in holoprosencephaly: interaction with the nuclear receptor NR4A3/NOR1. AB - Holoprosencephaly (HPE) is a relatively common brain malformation resulting in an incomplete separation of the two cerebral hemispheres. A number of mutations in different genes have been linked to this malformation, including three missense mutations in the homeodomain of the transcription factor SIX3. In this study, we investigated the functional consequences of these SIX3 mutations with respect to the ability of the protein to interact with and stimulate the transcriptional activity of the nuclear receptor NOR1 (NR4A3). Using glutathione S-transferase fusion protein pull-down assays and transient cotransfections of Neuro-2a cells with expression and reporter vectors, we found that one mutation, c.676C>G (p.L226V), does not alter the properties of SIX3 toward NOR1. Another mutation, c.749T>C (p.V250A), results in the production of a highly unstable protein in Neuro-2a cells. The third mutation, c.770G>C (p.R257P), results in a mutant SIX3 protein that no longer interacts with NOR1 in vivo. These observations suggest that different SIX3 mutations in HPE2 may affect different signaling pathways, and that one of these pathways may involve the nuclear receptor NOR1. PMID- 15523652 TI - Mutations in the MMAA gene in patients with the cblA disorder of vitamin B12 metabolism. AB - Mutations in the MMAA gene on human chromosome 4q31.21 result in vitamin B12 responsive methylmalonic aciduria (cblA complementation group) due to deficiency in the synthesis of adenosylcobalamin. Genomic DNA from 37 cblA patients, diagnosed on the basis of cellular adenosylcobalamin synthesis, methylmalonyl coenzyme A (CoA) mutase function, and complementation analysis, was analyzed for deleterious mutations in the MMAA gene by DNA sequencing of exons and flanking sequences. A total of 18 novel mutations were identified, bringing the total number of mutations identified in 37 cblA patients to 22. A total of 13 mutations result in premature stop codons; three are splice site defects; and six are missense mutations that occur at highly conserved residues. Eight of these mutations were common to two or more individuals. One mutation, c.433C>T (R145X), represents 43% of pathogenic alleles and a common haplotype was identified. Restriction endonuclease or heteroduplex diagnostic tests were designed to confirm mutations. None of the sequence changes identified in cblA patients were found in 100 alleles from unrelated control individuals. PMID- 15523657 TI - Further delineation of Frank-ter Haar syndrome. AB - The combination of the symptoms megalocornea, multiple skeletal anomalies, and developmental delay was first recognized as a separate entity by Frank et al. and subsequently confirmed by ter Haar et al. The main characteristics are brachycephaly, wide fontanels, prominent forehead, hypertelorism, prominent eyes, macrocornea with or without glaucoma, full cheeks, small chin, bowing of the long bones, and flexion deformity of the fingers. Protruding, simple ears, and prominent coccyx bone can be also regarded as important diagnostic signs. Inheritance most likely is autosomal recessive. Several manifestations such as progressive "coarsening" of the face, hirsutism, gallstones, lingual papillomatosis, and cardiac valve anomalies all point to a possible metabolic basis of the disorder. Here we describe four patients, including three sibs of Turkish descent, with this entity. PMID- 15523661 TI - Polycomb homologs are involved in teratogenicity of valproic acid in mice. AB - BACKGROUND: Valproic acid (VPA) is widely used to treat epilepsy and bipolar disorder and is also a potent teratogen, but its teratogenic mechanisms are unknown. We have attempted to describe a fundamental role of the Polycomb group (Pc-G) in VPA-induced transformations of the axial skeleton. METHODS: Pregnant NMRI mice were given a single subcutaneous injection of vehicle or VPA (800 mg/kg) on gestation day (GD) 8. The expression of genes encoding Polycomb and trithorax groups was measured by quantitative real-time RT-PCR using total RNA isolated from the embryos exposed to vehicle or VPA for 1, 3, and 6 hr. In addition, the use of two less teratogenic antiepileptic chemicals valpromide (VPD) and valnoctamide (VCD) provide reliable evidence to support the relationship between VPA teratogenicity and the Polycomb group. RESULTS: At a teratogenic level, VPA inhibits the expression of the Polycomb group genes, including Eed, Ezh2, Zfp144, Bmi1, Cbx2, Rnf2, and YY1 in the mouse embryos. In contrast, neither VPD nor VCD have significant effects on the expression of those genes affected by VPA. The trithorax group (trx-G) gene MLL, which is known to be required to maintain homeobox gene expression such as the Polycomb gene, is not affected by a teratogenic dose of VPA. CONCLUSIONS: We propose that, during embryonic development, VPA may affect the gene silencing pathway mediated by the Polycomb group complex. The epigenetic mechanism of VPA teratogenicity on anteroposterior patterning is suspected. PMID- 15523662 TI - Case for mandatory fortification of food with folate in Australia, for the prevention of neural tube defects. AB - BACKGROUND: Since the publication of randomized controlled trials demonstrating the prevention of neural tube defects (NTDs) with periconceptional folate, several Australian states have promoted an increase in periconceptional use of folic acid supplements. Since 1996, voluntary fortification of food with folate has been allowed in Australia and New Zealand for the purpose of preventing NTDs. METHODS: For this report, we synthesized published and unpublished data on folic acid supplement use, voluntary fortification, and trends in NTDs. RESULTS: There has been an increase in the proportion of women (up to 30-40%) taking periconceptional folic acid supplements in Australia, and many foods (mainly breakfast cereals) are fortified. Supplement use is strongly correlated with educational and socioeconomic status; consumption of voluntarily fortified foods is not. There has been a fall in NTDs of about 30% in the non-Aboriginal population, but no change has been seen in the Aboriginal population. CONCLUSIONS: These data support mandatory fortification of food as a more equitable approach to achieving sufficient folate intake in the periconceptional period for all women in Australia and New Zealand to prevent the majority of NTDs in their offspring. In May 2004, based on these and other considerations, the Australia and New Zealand Food Regulation Ministerial Council agreed that mandatory fortification of food with folate should be considered as a priority. PMID- 15523663 TI - Hungarian cohort-controlled trial of periconceptional multivitamin supplementation shows a reduction in certain congenital abnormalities. AB - BACKGROUND: The 1984-1991 Hungarian randomized controlled trial (RCT) of periconceptional multivitamin supplementation containing folic acid (0.8 mg) showed a significant reduction in the first occurrence of neural tube defects (NTDs), and of urinary tract and cardiovascular abnormalities, but no reduction in orofacial clefts. A controlled cohort trial was designed to confirm or deny these results. METHODS: Supplemented women were recruited from the Hungarian Periconceptional Service using the same multivitamin as the Hungarian RCT. Unsupplemented pregnant women were recruited in the standard regional antenatal care clinics and were matched to each supplemented pregnant woman on the basis of age, socioeconomic status, place of residence, and year of pregnancy. RESULTS: A total of 3056 informative offspring were evaluated in each cohort. The occurrence of congenital cardiovascular malformations (31 vs. 50) was reduced (odds ratio [OR], 0.60; 95% confidence interval [CI], 0.38-0.96) in the supplemented cohort, accounted for mainly by ventricular septal defects (5 vs. 19; OR, 0.26; 95% CI, 0.09-0.72). There was no significant difference (14 vs. 19) in the occurrence of urinary tract defects between the two cohorts, but stenosis/atresia of pelvic ureteric junction (2 vs. 13) showed a significant reduction (OR, 0.19; 95% CI, 0.04-0.86). The protective effect of the folic acid-containing multivitamin for NTDs (one offspring in the supplemented vs. nine in the unsupplemented cohort) was confirmed (OR, 0.11; 95% CI, 0.01-0.91). There was, however, no protective effect on orofacial clefts or on multiple congenital abnormalities. CONCLUSIONS: The results of this cohort-controlled trial support the findings of the previous Hungarian RCT. The primary prevention of some major structural birth defects by multivitamins containing folic acid or by folic acid has great public health importance. PMID- 15523664 TI - Periconceptional multivitamin intake during early pregnancy, genetic variation of acetyl-N-transferase 1 (NAT1), and risk for orofacial clefts. AB - BACKGROUND: Periconceptional supplementation of multivitamins that include folic acid have been shown to prevent several birth defects, including neural tube defects and orofacial clefts. We investigated whether polymorphic variants of fetal acetyl-N-transferase 1 (NAT1), an enzyme involved in the catabolism of folates, differentially interacted with maternal multivitamin use during early pregnancy to alter the risk of delivering an infant with an orofacial cleft malformation. METHODS: Using a large population-based case-control study, we genotyped 421 California infants born with an isolated cleft and 299 controls for two NAT1 polymorphisms. RESULTS: Compared to the homozygous wild-type genotypes, odds ratios for isolated cleft lip with/without cleft palate were slightly increased among infants who were homozygous for the variant alleles of NAT1 1088 and 1095. For isolated cleft palate, no similar associations with these two NAT1 variants were observed. For NAT1 1088 genotypes, we did not observe any differential risks for clefts related to maternal multivitamin intake. For NAT1 1095 genotypes, however, we found a two-fold higher risk for isolated cleft lip with/without cleft palate among infants who were homozygous for the variant allele and whose mothers did not take multivitamins during early pregnancy. CONCLUSIONS: We found evidence suggestive of an interaction between the NAT1 1095 polymorphism and lack of maternal multivitamin use that increased risks of isolated cleft lip with/without cleft palate. PMID- 15523665 TI - Encephalocraniocutaneous lipomatosis accompanied by maxillary compound odontoma and juvenile angiofibroma: report of a case. AB - BACKGROUND: Encephalocraniocutaneous lipomatosis (ECCL) is a rare neurocutaneous syndrome characterized by congenital cutaneous, ocular, and neurologic abnormalities, which may be pronounced in the head and neck. CASE: In this article we present a well-documented case of a six-year-old boy with ECCL associated with oral manifestations. In this case, typical features such as large lipomatous brown pigmented plaques of the top of the skull with overlying alopecia, ptotic eyelid with decreased function, bulbar conjunctiva lipodermoid, microcalcifications and atrophy of cerebral parenchyma, and the widening of the frontal subarachnoid space and the fissure of Sylvius were accompanied by intraoral lesions, maxillary compound odontoma, and juvenile extranasopharyngeal angiofibroma of the gingiva. CONCLUSIONS: Although cases of odontomas have already been described and angiofibromas are a quite common extraoral finding, to our knowledge this is the first case of intraoral evaluation of both extranasopharyngeal juvenile angiofibroma and maxillary compound odontoma in ECCL syndrome. PMID- 15523667 TI - Principal eigenvector of contact matrices and hydrophobicity profiles in proteins. AB - With the aim of studying the relationship between protein sequences and their native structures, we adopted vectorial representations for both sequence and structure. The structural representation was based on the principal eigenvector of the fold's contact matrix (PE). As has been recently shown, the latter encodes sufficient information for reconstructing the whole contact matrix. The sequence was represented through a hydrophobicity profile (HP), using a generalized hydrophobicity scale that we obtained from the principal eigenvector of a residue residue interaction matrix, and denoted as interactivity scale. Using this novel scale, we defined the optimal HP of a protein fold, and, by means of stability arguments, predicted to be strongly correlated with the PE of the fold's contact matrix. This prediction was confirmed through an evolutionary analysis, which showed that the PE correlates with the HP of each individual sequence adopting the same fold and, even more strongly, with the average HP of this set of sequences. Thus, protein sequences evolve in such a way that their average HP is close to the optimal one, implying that neutral evolution can be viewed as a kind of motion in sequence space around the optimal HP. Our results indicate that the correlation coefficient between N-dimensional vectors constitutes a natural metric in the vectorial space in which we represent both protein sequences and protein structures, which we call vectorial protein space. In this way, we define a unified framework for sequence-to-sequence, sequence-to-structure and structure to-structure alignments. We show that the interactivity scale is nearly optimal both for the comparison of sequences to sequences and sequences to structures. PMID- 15523666 TI - Fold recognition by combining sequence profiles derived from evolution and from depth-dependent structural alignment of fragments. AB - Recognizing structural similarity without significant sequence identity has proved to be a challenging task. Sequence-based and structure-based methods as well as their combinations have been developed. Here, we propose a fold recognition method that incorporates structural information without the need of sequence-to-structure threading. This is accomplished by generating sequence profiles from protein structural fragments. The structure-derived sequence profiles allow a simple integration with evolution-derived sequence profiles and secondary-structural information for an optimized alignment by efficient dynamic programming. The resulting method (called SP(3)) is found to make a statistically significant improvement in both sensitivity of fold recognition and accuracy of alignment over the method based on evolution-derived sequence profiles alone (SP) and the method based on evolution-derived sequence profile and secondary structure profile (SP(2)). SP(3) was tested in SALIGN benchmark for alignment accuracy and Lindahl, PROSPECTOR 3.0, and LiveBench 8.0 benchmarks for remote homology detection and model accuracy. SP(3) is found to be the most sensitive and accurate single-method server in all benchmarks tested where other methods are available for comparison (although its results are statistically indistinguishable from the next best in some cases and the comparison is subjected to the limitation of time-dependent sequence and/or structural library used by different methods.). In LiveBench 8.0, its accuracy rivals some of the consensus methods such as ShotGun-INBGU, Pmodeller3, Pcons4, and ROBETTA. SP(3) fold-recognition server is available on http://theory.med.buffalo.edu. PMID- 15523668 TI - Predicting absolute contact numbers of native protein structure from amino acid sequence. AB - The contact number of an amino acid residue in a protein structure is defined by the number of C(beta) atoms around the C(beta) atom of the given residue, a quantity similar to, but different from, solvent accessible surface area. We present a method to predict the contact numbers of a protein from its amino acid sequence. The method is based on a simple linear regression scheme and predicts the absolute values of contact numbers. When single sequences are used for both parameter estimation and cross-validation, the present method predicts the contact numbers with a correlation coefficient of 0.555 on average. When multiple sequence alignments are used, the correlation increases to 0.627, which is a significant improvement over previous methods. In terms of discrete states prediction, the accuracies for 2-, 3-, and 10-state predictions are, respectively, 71.4%, 54.1%, and 18.9% with residue type-dependent unbiased thresholds, and 76.3%, 59.2%, and 21.8% with residue type-independent unbiased thresholds. The difference between accessible surface area and contact number from a prediction viewpoint and the application of contact number prediction to three-dimensional structure prediction are discussed. PMID- 15523670 TI - In support of fatty acid synthase (FAS) as a metabolic oncogene: extracellular acidosis acts in an epigenetic fashion activating FAS gene expression in cancer cells. AB - Relatively little information exists on the ultimate molecular mechanisms by which the lipogenic enzyme Fatty Acid Synthase (FAS) is differentially overexpressed in a biologically aggressive subset of human malignancies. Since the microenvironment of solid tumors contains regions of poor oxygenation and high acidity, it has recently been suggested that cancer-associated FAS is a novel metabolic oncogene conferring a selective growth advantage upon stresses such as hypoxia and/or low pH. Here, we performed transient transfection studies with a 178-bp FAS promoter fragment harboring a complex Sterol Regulatory Element Binding Proteins (SREBP)-binding site to evaluate whether extracellular low pH and/or hypoxia may act in an epigenetic fashion by inducing changes in the transcriptional activation of FAS gene in cancer cells. First, MCF-7 breast cancer cells cultured in acidosis (pH 6.5), but not under hypoxia or in the presence of hypoxia mimetics, demonstrated a more than two-fold increase in the transcriptional activity of FAS promoter-reporter constructs compared with control cells grown under standard culture conditions (pH 7.4). Second, the up regulatory effect of extracellular acidosis on the transcriptional activation of FAS gene was not observed when the FAS promoter was truncated at the SREBP binding site. Third, MCF-7 cells engineered to overexpress the Her-2/neu (erbB-2) oncogene exhibited a SREBP-dependent activation of the FAS promoter-reporter construct up to three-fold higher than that found in wild-type MCF-7 cells, while extracellular acidosis resulted only in a marginal increase of Her-2/neu-promoted activation of FAS gene. This study reveals for the first time that extracellular acidosis can work in an epigenetic fashion by up-regulating the transcriptional expression of FAS gene in breast cancer cells, a stimulatory effect that is equally mimicked by well-characterized oncogenic stimuli such as Her-2/neu. These findings, altogether, support the "metabolic oncogene" theory for FAS overexpression in cancer cells. PMID- 15523671 TI - Electron microscopy of DNA replication in 3-D: evidence for similar-sized replication foci throughout S-phase. AB - DNA replication sites (RS) in synchronized HeLa cells have been studied at the electron microscopic level. Using an improved method for detection following the in vivo incorporation of biotin-16-deoxyuridine triphosphate, discrete RS, or foci are observed throughout the S-phase. In particular, the much larger RS or foci typically observed by fluorescence microscopic approaches in mid- and late-S phase, are found to be composed of smaller discrete foci that are virtually identical in size to the RS observed in early-S-phase. Pulse-chase experiments demonstrate that the RS of early-S-phase are maintained when chased through S phase and into the next cell generation. Stereologic analysis demonstrates that the relative number of smaller sized foci present at a given time remains constant from early through mid-S-phase with only a slight decrease in late-S phase. 3-D reconstruction of serial sections reveals a network-like organization of the RS in early-S-phase and confirms that numerous smaller-sized replication foci comprise the larger RS characteristic of late-S-phase. PMID- 15523672 TI - Specificity of transcriptional regulation by the zinc finger transcription factors Sp1, Sp3, and Egr-1. AB - The transcription factors Sp1, Sp3, and Egr-1 bind with their zinc finger DNA binding domains to GC-rich sequences in the regulatory regions of their target genes. The similarity of the DNA-binding sites of Sp1, Sp3, and Egr-1 has triggered the hypothesis that they compete for the same DNA-binding site. We have investigated the specificity of transcriptional regulation by Sp1, Sp3, and Egr-1 using dominant-negative mutants that block the DNA-binding site of Sp1, Sp3, or Egr-1, respectively. The results show that constitutive transcription of Sp1 regulated reporter genes, containing Sp1 sites derived from the aldolase C and p21WAF1/Cip1 genes, or the long terminal repeat of HIV-1, was impaired by dominant-negative mutants of Sp1 and Sp3, but not by a dominant-negative Egr-1. Transcription mediated by Egr-1 was induced by transfection of expression vectors encoding wild-type or mutated Egr-1 or by stimulation of the extracellular signal regulated protein kinase pathway via an inducible B-Raf-estrogen receptor fusion protein. In all cases transcription of Egr-1-regulated reporter genes, containing Egr-1 binding sites derived from the Egr-1 or the synapsin I gene was impaired by a dominant-negative Egr-1, but not by dominant-negative Sp1 or Sp3 mutants. These results show that there are genuine Sp1/Sp3 or Egr-1 controlled genes showing no cross-regulation of Sp1/Sp3 and Egr-1 through the same DNA-binding site. This does not exclude the existence of composite Sp1/Sp3/Egr-1 binding sites, where competition for a common DNA-binding site occurs. PMID- 15523674 TI - Regulation of gene expression in melanoma: new approaches for treatment. AB - The molecular changes associated with the transition of melanoma cells from radial growth phase (RGP) to vertical growth phase (VGP, metastatic phenotype) are not yet well defined. We have demonstrated that the progression of human melanoma is associated with loss of expression of the transcription factor AP-2. In metastatic melanoma cells, this loss resulted in overexpression of MCAM/MUC18, MMP-2, the thrombin receptor (PAR-1), and lack of c-KIT expression. The transition from RGP to VGP is also associated with overexpression of the angiogenic factor IL-8. Additionally, the transition of melanoma cells from RGP to VGP is associated with overexpression of the transcription factors CREB and ATF-1, both of which may act as survival factors for human melanoma cells. Inactivation of CREB/ATF-1 activities in metastatic melanoma cells by dominant negative CREB or by anti-ATF-1 single chain antibody fragment (ScFv), resulted in deregulation of MMP-2 and MCAM/MUC18, increased the sensitivity of melanoma cells to apoptosis, and inhibition of their tumorigenicity and metastatic potential in vivo. In this prospect article, we summarize our data on the role of AP-2 and CREB/ATF-1 in the progression of human melanoma and report on the development of new fully human antibodies anti-MCAM/MUC18 and anti-IL-8 which could serve as new modalities for the treatment of melanoma. PMID- 15523673 TI - Structural and functional analysis of domains mediating interaction between NKX 3.1 and PDEF. AB - NKX-3.1 is a suspected prostate tumor suppressor gene that encodes a homeodomain transcription factor. NKX-3.1 has been demonstrated to interact with prostate derived Ets factor (PDEF) and to suppress the ability of PDEF to transactivate the prostate specific antigen promoter. To dissect the molecular basis of the interaction between these transcription factors, deletion analyses were preformed using the yeast two-hybrid system. The interaction of NKX-3.1 with full-length PDEF requires part of the homeodomain and a tyrosine-rich 21 amino acid sequence that lies C-terminal to the homeodomain. The interaction of PDEF with full-length NKX-3.1 requires the Ets domain and a linker region that lies between the Ets and pointed domains. Deletion of the C-terminal 21 amino acids of NKX-3.1 completely disrupts the ability to suppress the transactivation function of PDEF in prostate tumor cells, demonstrating concordance between interaction in yeast and function in mammalian cells. These studies have identified novel protein-protein interaction domains within NKX-3.1 and PDEF that operate in concert with their respective DNA binding domains to mediate functional interactions between these growth regulatory transcription factors. PMID- 15523675 TI - Modulation of 1alpha,25-dihydroxyvitamin D3-membrane associated, rapid response steroid binding protein expression in mouse odontoblasts by 1alpha,25-(OH)2D3. AB - The rapid, nongenomic effects of 1alpha,25-dihydroxyvitamin D3 (1alpha,25-(OH)2D3 have been related to a 1,25D3-membrane associated, rapid response steroid binding protein or 1,25D3-[MARRS]bp, with a molecular weight of 65 kDa, in several tissues and species. Currently, no information is available concerning the nongenomic responses to 1alpha,25-(OH)2D3 in dental tissues. In order to investigate the expression of 1,25D3-[MARRS]bp in dental cells, in the presence or absence of 1alpha,25-(OH)2D3, we have used rabbit polyclonal antibodies directed against the N-terminus of the 1,25D3-[MARRS]bp (Ab099) that recognizes the 1alpha,25-(OH)2D3 binding protein in chick intestinal basolateral membranes and a mouse odontoblast-like cell line (MO6-G3). Western blotting and flow cytometric analyses with Ab099 specifically detected 1,25D3-[MARRS]bp in MO6-G3 cells. Moreover, 1,25D3-[MARRS]bp was up-regulated, in vivo, in differentiated dental cells. Electron microscopic analysis confirmed the plasma membrane localization of this binding protein and also showed its intracellular presence. Incubation of MO6-G3 cells with different doses of 1alpha,25-(OH)2D3 for 36 h resulted in an inhibition of 1,25D3-[MARRS]bp expression with a maximal effect at 50 nM steroid. In addition, the culture media of MO6-G3 cells contains immunoreactive 1,25D3-[MARRS]bp. Immunogold positive membrane vesicle-like structures are present in the extracellular matrix of MO6-G3 cells. Altogether, these results indicate that the 1,25D3-[MARRS]bp expression in MO6-G3 cells is modulated by 1alpha,25-(OH)2D3. In conclusion, this 1alpha,25-(OH)2D3 binding protein could play an important role in the rapid, nongenomic responses to 1alpha,25-(OH)2D3 in dental cells. PMID- 15523676 TI - Determination of structural and functional overlap/divergence of five proto-type galectins by analysis of the growth-regulatory interaction with ganglioside GM1 in silico and in vitro on human neuroblastoma cells. AB - The growth-regulatory interplay between ganglioside GM1 on human SK-N-MC neuroblastoma cells and an endogenous lectin provides a telling example for glycan (polysaccharide) functionality. Galectin-1 is the essential link between the sugar signal and the intracellular response. The emerging intrafamily complexity of galectins raises the question on defining extent of their structural and functional overlap/divergence. We address this problem for proto type galectins in this system: ganglioside GM1 as ligand, neuroblastoma cells as target. Using the way human galectin-1 interacts with this complex natural ligand as template, we first defined equivalent positioning for distinct substitutions in the other tested proto-type galectins, e.g., Lys63 vs. Leu60/Gln72 in galectins-2 and -5. As predicted from our in silico work, the tested proto-type galectins have affinity for the pentasaccharide of ganglioside GM1. In contrast to solid-phase assays, cell surface presentation of the ganglioside did not support binding of galectin-5, revealing the first level of regulation. Next, a monomeric proto-type galectin (CG-14) can impair galectin-1-dependent negative growth control by competitively blocking access to the shared ligand without acting as effector. Thus, the quaternary structure of proto-type galectins is an efficient means to give rise to functional divergence. The identification of this second level of regulation is relevant for diagnostic monitoring. It might be exploited therapeutically by producing galectin variants tailored to interfere with galectin activities associated with the malignant phenotype. Moreover, the given strategy for comparative computational analysis of extended binding sites has implications for the rational design of galectin-type-specific ligands. PMID- 15523677 TI - Microwave fabrication of MFI zeolite crystals with a fibrous morphology and their applications. PMID- 15523678 TI - Cubic, hydrogen-bonded (10,3)-a networks in the family [C(NH2)3][N(CH3)4][XO4] (X = S, Cr, and Mo). PMID- 15523679 TI - Primo Levi's The Periodic Table. A search for patterns in times past. PMID- 15523680 TI - Catalytic promiscuity in biocatalysis: using old enzymes to form new bonds and follow new pathways. AB - Biocatalysis has expanded rapidly in the last decades with the discoveries of highly stereoselective enzymes with broad substrate specificity. A new frontier for biocatalysis is broad reaction specificity, where enzymes catalyze alternate reactions. Although often under-appreciated, catalytic promiscuity has a natural role in evolution and occasionally in the biosynthesis of secondary metabolites. Examples of catalytic promiscuity with current or potential applications in synthesis are reviewed here. Combined with protein engineering, the catalytic promiscuity of enzymes may broadly extend their usefulness in organic synthesis. PMID- 15523681 TI - Are NMR-derived model structures for beta-peptides representative for the ensemble of structures adopted in solution? PMID- 15523682 TI - Novel inhibitors targeted to methionine aminopeptidase 2 (MetAP2) strongly inhibit the growth of cancers in xenografted nude model. AB - Inhibition of angiogenesis is emerging as a promising strategy for the treatment of cancer. In our study reported here, the effects of 4 highly potent methionine aminopeptidase 2 (MetAP2) inhibitors, IDR-803, IDR-804, IDR-805 and CKD-732 (designed by structure-based molecular modeling), on angiogenesis and tumor growth were assessed. Concentrations of these inhibitors as low as 2.5 nM were able to inhibit the growth of human umbilical vein endothelial cells (HUVEC) by as much as 50%, arresting growth in the G1 stage of mitosis. An intracellular accumulation of p21(WAF1/Cip1) protein was also observed. Furthermore, at higher concentrations (25 nM) of these 4 MetAP2 inhibitors, a significant induction of apoptosis was apparent in the same HUVEC cultures. As a result of these findings, the possible anticancer effects of these inhibitors were examined, utilizing the SNU-398 hepatoma cell line. Interestingly, pretreatment with these inhibitors led to an increased number of apoptotic cells of up to 60% or more, compared to untreated controls. Moreover, utilizing an in vivo xenografted murine model, these inhibitors suppressed the growth of engrafted tumor. In conclusion, these 4 inhibitory compounds potently exert an antiangiogenic effect to inhibit the growth of cancers in vivo and could potentially be useful for the treatment of a variety of cancers. PMID- 15523683 TI - Magnetic resonance imaging in an orthotopic rat model: blockade of epidermal growth factor receptor with EMD72000 inhibits human pancreatic carcinoma growth. AB - The purpose of our research was to investigate the antiangiogenic effect of the epidermal growth factor receptor monoclonal antibody (anti-EGF-R MAB) EMD72000, in an orthotopic human pancreatic carcinoma model in rats, assessed by magnetic resonance (MR) imaging using angiogenic surrogate markers in comparison with histopathologic findings. Human pancreatic adenocarcinoma cells L3.6pl were injected orthotopically in the pancreas of 12 athymic nude rats. Through a 21-day course, groups of 6 rats were treated intraperitoneally with either EMD72000 or with saline solution for control animals. Dynamic contrast-enhanced MR imaging was performed before and after the treatment to assess microvascular permeability, estimated by the endothelial transfer coefficient (KPS) and fractional plasma volumes (fPV) of the pancreatic tumors. EMD72000-treated animals showed significantly less tumor volume progression (1,080 mm3 +/- 1,244; p = 0.012) and significantly lower values for microvascular permeability (KPS = 4.2 ml min(-1) 100 ml(-1) of tissue +/- 2.8; p = 0.015), fractional plasma volume (fPV = 0.018 ml ml(-1) of tissue +/- .015; p = 0.003) and microvessel density (MVD = 13 +/- 4 (0.159 mm2); p = 0.001) than saline-treated animals (6,544 mm3 +/ 5,202; 9.5 ml min(-1) 100 ml(-1) of tissue +/- 4.3, 0.056 ml ml(-1) of tissue +/ 0.019 and 25 +/- 5 (0.159 mm2), respectively). KPS and fPV values showed moderate positive correlation with MVD (r = 0.5, p = 0.103; r = 0.6, p = 0.065, respectively). Intraperitoneal injection of EMD72000 inhibits orthotopic human pancreatic carcinoma growth in rats. Antiangiogenic effects of anti-EGF-R MAB EMD72000 can be quantified and monitored noninvasively by dynamic MR imaging. PMID- 15523684 TI - A novel NF-kappaB inhibitor DHMEQ selectively targets constitutive NF-kappaB activity and induces apoptosis of multiple myeloma cells in vitro and in vivo. AB - Multiple myeloma (MM) is a fatal lymphoid malignancy that is incurable with conventional modalities of chemotherapy. Strong and constitutive activation of nuclear factor kappa B (NF-kappaB) is a common characteristic of MM cells. In our study we successfully target NF-kappaB with a novel NF-kappaB inhibitor dehydroxymethylepoxyquinomycin (DHMEQ). DHMEQ completely abrogates constitutive NF-kappaB activity and induces apoptosis of MM cells, whereas control peripheral blood mononuclear cells (PBMC) are resistant to NF-kappaB inhibition and apoptosis by DHMEQ treatment. DHMEQ inhibition of NF-kappaB triggers activation of caspases 8 and 9, as well as G0/G1 cell cycle arrest accompanied by downregulation of antiapoptotic genes Bcl-XL and c-FLIP and cell cycle progression gene cyclins D1 and D2. DHMEQ-mediated inhibition of vascular endothelial growth factor (VEGF) production in MM cells raises the possibility that DHMEQ abrogates the autocrine VEGF loop and enhances its antitumor effects by inhibiting neovascularization in the bone marrow. Using an in vivo NOD/SCID/gammac(null) (NOG) mice model, we show that DHMEQ has a potent inhibitory effect on the growth of MM cells. Compared to other compounds having the potential to inhibit NF-kappaB, DHMEQ is a unique compound that blocks the translocation of NF-kappaB p65 into the nucleus and selectively targets NF-kappaB activated in tumor cells. Therefore, our study presents a new molecular target therapy in MM. PMID- 15523685 TI - p53 autoantibodies predict subsequent development of cancer. AB - Because TP53 mutations can induce an immune response and can occur early in the carcinogenic process for some tumors, p53 autoantibodies may be useful biomarkers for risk of development of cancer. Using banked serum samples from an asbestosis cohort at high risk for cancer, we demonstrate for the first time a statistically significant relationship between p53 autoantibodies and the subsequent development of malignancy (hazard ratio [HR] = 5.5, 95% confidence interval [CI] = 2.8-10.9) with a positive predictive value of 0.76 and an average lead time to diagnosis of 3.5 years. p53 autoantibodies were also significantly associated with p53 alterations in the resultant tumors (kappa = 0.78, p = 0.01). PMID- 15523686 TI - Oxidative DNA damage and antioxidant vitamin level: comparison among lung cancer patients, healthy smokers and nonsmokers. AB - In the present study, we examined whether the level of 8-oxo-7,8-dihydro-2' deoxyguanosine (8-oxodGuo) in leukocyte DNA is higher in lung cancer patients compared to controls. Factors that may influence oxidative stress, such as antioxidant vitamins, were also determined. These parameters were analyzed in 4 groups of subjects: smokers with lung cancer, ex-smokers with lung cancer, healthy smokers with comparable smoking status and healthy nonsmokers. The 8 oxodGuo mean level in leukocytes of lung cancer patients reached values of 9.22/10(6) dGuo molecules (smokers) and 11.16/10(6) dGuo molecules (ex-smokers). These values were significantly higher than in DNA of healthy smokers and nonsmokers, where mean levels reached 6.99/10(6) dGuo molecules and 5.98/10(6) dGuo molecules, respectively. Mean levels of vitamin C in the plasma of controls and lung cancer patients were 56.17 microM (nonsmokers), 26.34 microM (healthy smokers), 23.83 microM (cancer patients, smokers) and 29.19 microM (cancer patients, ex-smokers). The difference between nonsmokers and the 3 other groups was statistically significant. Vitamin E level was significantly reduced in the plasma of cancer patients (smokers 19.94 microM, ex-smokers 19.59 microM) compared to healthy smokers (28.93 microM). No changes in vitamin A concentration were found. Our results suggest that a high level of 8-oxodGuo in leukocyte DNA and a low concentration of vitamin E in the blood may predict lung cancer risk. However, it is also possible that these phenomena may simply result from disease development. PMID- 15523687 TI - Ursolic acid induces apoptosis through mitochondrial intrinsic pathway and caspase-3 activation in M4Beu melanoma cells. AB - Over the coming years, skin cancer could become a significant public health problem. Previous results indicate that ursolic acid (UA), a pentacyclic triterpene acid, has pleiotropic biologic activities such as antiinflammatory and antiproliferative activities on cancer cells. As UA represents a promising chemical entity for the protection of human skin, in agreement with tests done by the cosmetic industry, we investigated its effects on the M4Beu human melanoma cell line. In this report, we demonstrated for the first time that UA had a significant antiproliferative effect on M4Beu, associated with the induction of an apoptotic process, characterized by caspase-3 activation, the downstream central effector of apoptosis. We demonstrated that UA-induced apoptosis was dependent on the mitochondrial intrinsic pathway, as shown by transmembrane potential collapse (DeltaPsim) and by alteration of the Bax-Bcl-2 balance, with a concomitant increase in Bax expression and decrease in Bcl-2 expression. We also showed that UA-induced DeltaPsim was associated with apoptosis-inducing factor leakage from mitochondria. Taken together, our results suggest that UA-induced apoptosis on M4Beu cells is accomplished via triggering of mitochondrial pathway. In conclusion, UA could be an encouraging compound in the treatment or prevention of skin cancer and may represent a new promising anticancer agent in the treatment of melanoma. PMID- 15523688 TI - SEREX identification of new tumor antigens linked to melanoma-associated retinopathy. AB - Metastatic melanoma still has a very poor prognosis since it withstands conventional therapies like surgery or chemotherapy. A paraneoplastic autoimmune manifestation of this disease is melanoma-associated retinopathy (MAR). MAR has been associated with prolonged survival and may be an early marker of tumor progression. By screening a retina and a melanoma cDNA phage library by SEREX using sera of patients suffering from melanoma and, in some cases, clinical symptoms of MAR, we identified 20 new antigens (HD-MM-28-47), of which 14 clones had high homology to well-known genes. Six of these genes had previously been associated with retina: rhodopsin, visual arrestin, MEK1, SRPX, BBS1 and galectin 3. Individual clones were recognized by up to 43% of patients' sera, while sera of healthy volunteers were negative except in 2 cases. The expression profile of the antigens identified on the basis of homologous EST database entries in healthy tissues was ubiquitous to differential. Using RT-PCR, we found frequent expression of preselected antigens in melanoma cell lines. For rhodopsin, this could be quantified by quantitative PCR. Retinal proteins were recognized by serum antibodies of melanoma patients but not healthy controls. The role of these antigens in MAR awaits further investigation. (Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020-7136/suppmat/index.html.) PMID- 15523689 TI - Expression of NGF in hepatocellular carcinoma cells with its receptors in non tumor cell components. AB - Nerve growth factor (NGF) is suggested to have a role in tumor progression in addition to its role in differentiation and survival of neuronal cells. We investigated expression of NGF and its receptors, TrkA and p75NTR, in hepatocellular carcinomas (HCCs). Although hepatocytes and hepatic stellate cells (HSCs) showed respectively weak and intense NGF immunostaining in the background livers of patients suffering from liver cirrhosis (LC) or chronic hepatitis (CH), intense staining was demonstrated in HCC cells of 33 of 54 (61.1%) tumors. RT-PCR detected NGF mRNA in 7 freshly-isolated HCC samples, and in 2 of 4 cases, in which both background livers and tumors could be analyzed, NGF mRNA was more abundant in the tumors than the background livers. TrkA was detected in the smooth muscle cells of hepatic arteries, but it was negative in tumor cells as well as non-neoplastic hepatocytes. p75NTR and alpha-smooth muscle actin (alphaSMA) was expressed in HSCs in the background liver and fibroblast-like cells in stromal septa, whereas HSCs within the HCC tissues were mostly negative for p75NTR but positive for alphaSMA. This suggests that HSCs in HCC have a different property from those in background livers. Furthermore, the stromal septa contained abundant nerve fibers, which may be related to the increased NGF expression in HCC cells. NGF and its receptors are then thought to have a role in cellular interactions involving HCC cells, HSCs, arterial cells and nerve cells in HCC tissues. PMID- 15523690 TI - DHPLC is superior to SSCP in screening p53 mutations in esophageal cancer tissues. AB - Mutations of the p53 tumor-suppressor gene universally occur on exons 5-8 in human cancer. We analyzed these mutations in esophageal cancer tissue from 207 patients in China using 2 methods, single-strand conformation polymorphism (SSCP), one of the most frequently used methods, and the recently developed denaturing high-performance liquid chromatography (DHPLC), and compared their sensitivity and efficiency. Exons 5-8 of p53 were amplified from esophageal cancer tissue genomes, screened for fragments of mutations and polymorphisms by SSCP and DHPLC in a blind study and confirmed by direct sequencing to detect the mutations and polymorphisms. The numbers detected by DHPLC were greater than those detected by SSCP, though the rate of mutations and polymorphisms was lower in SSCP than in DHPLC, which appeared to detect smaller mutations (substitutions and 1 bp insertions/deletions). Of the mutations with substitutions detected by DHPLC but not by SSCP, 50% substituted adenosine for other nucleotides, suggesting that these mutations are often missed when SSCP is used. According to these data, the sensitivity of SSCP and DHPLC was 81% and 97%, respectively, and the specificity was 97% and 85%, respectively. Our results suggest that DHPLC may be recommended over SSCP when screening gene mutations. Thus, rates of p53 mutations and polymorphisms in esophageal cancer tissue in Chinese patients were 49% and 41% by DHPLC and SSCP, respectively. PMID- 15523691 TI - Depressed tumor necrosis factor alpha and interleukin-12p40 production by peripheral blood mononuclear cells of gastric cancer patients: association with IL-1R-associated kinase-1 protein expression and disease stage. AB - Our study investigated the ability of peripheral blood mononuclear cells (PBMCs) isolated from patients with different clinical stages of gastric cancer to produce proinflammatory (tumor necrosis factor alpha [TNFalpha], interleukin 12p40 [IL-12p40] and interleukin 6 [IL-6]) and antiinflammatory (interleukin-10 [IL-10]) cytokines after stimulation with lipopolysaccharide (LPS) or tumor cells, and its correlation with IL-1R-associated kinase-1 (IRAK-1) protein expression. The data showed that TNF production by tumor cell-stimulated PBMCs obtained from patients with advanced gastric cancer was significantly depressed in comparison to the control group. The response to LPS was less affected. IL 12p40 production was depressed in all stages of disease, while the release of IL 10 and IL-6 remained unchanged. Depressed tumor cell-induced TNF and IL-12p40 production was associated with diminished IRAK-1 protein expression in PBMC. These findings may suggest that in advanced gastric cancer (at least in some cancer patients) diminished IRAK-1 protein expression may be a novel mechanism responsible for or facilitating downregulation of innate immune response to tumor cells. PMID- 15523692 TI - Unmasking immunosurveillance against a syngeneic colon cancer by elimination of CD4+ NKT regulatory cells and IL-13. AB - We have previously observed a novel role of natural killer T (NKT) cells in negative regulation of antitumor immune responses against an immunogenic regressor tumor expressing a transfected viral antigen. Here, we investigated whether hidden spontaneous antitumor immunosurveillance, in the absence of a vaccine, could be revealed by disruption of this negative regulatory pathway involving CD4+ NKT cells and interleukin-13 (IL-13), in a murine pulmonary metastasis model of a nontransfected, nonregressor, syngeneic tumor, the CT26 colon carcinoma. Lung metastases of CT26 were decreased in CD4+ T cell-depleted BALB/c mice, suggesting that CD4+ T cells were involved in negative regulation of antitumor responses. CD1-knock out (CD1-KO) mice, which have conventional CD4+ T cells and CD4+CD25+ regulatory T cells but lack CD1-restricted CD4+ NKT cells, were significantly resistant to lung metastasis of CT26. The metastases were not further decreased in CD4+ T cell-depleted CD1-KO mice, implying that CD4+ NKT cells might be the primary negative regulator of antitumor immune responses in BALB/c mice. CD8+ T cells were found to act as effectors in antitumor immune responses, since the inhibition of lung metastases observed in naive CD1-KO or CD4+ T cell-depleted mice was abrogated by depletion of CD8+ T cells. Lung metastases were significantly decreased by treatment of mice with an IL-13 inhibitor, but not by deficiency or inhibition of IL-4. Thus, even for a nonregressor tumor, immunosurveillance exists but is negatively regulated via CD4+ NKT cells possibly mediated by IL-13, and can be unmasked by removal of these negative regulatory components. PMID- 15523693 TI - Flavonoid apigenin inhibits motility and invasiveness of carcinoma cells in vitro. AB - Investigations of the mechanisms of the cancer-preventive activity of apigenin (4',5,7,-trihydroxyflavone), a plant-derived, anti-carcinogenic flavonoid, showed its interference with cell proliferation, survival, and gap junctional coupling. We used a model based on non-invasive HeLa wild-type cells and their connexin43 (Cx43) transfected counterparts to correlate the effect of apigenin on tumour cell invasiveness with its influence on cell motility. Both cell lines displayed similar motile properties in control conditions. Apigenin treatment resulted in a significant and reversible inhibition of translocation of both HeLa wild-type cells and HeLa Cx43 transfectants. The effect of apigenin on cell proliferation was less pronounced especially at low apigenin concentration, whereas its influence on cell motility correlated with the reduction of the invasive potential of HeLa Cx43 cells as shown by an invasion assay based on the confrontation of tumour cell spheroids with chick embryo heart fragments. HeLa Cx43 cells were highly invasive in controls, but did not invade the heart tissue at tumour cell aggregate-fibroblast capsule interfaces in the presence of apigenin and failed to fully engulf these heart fragments. Because the motility of chick heart fibroblasts was only slightly affected by apigenin, these observations indicate that apigenin exerts its anti-invasive effect on HeLa cells predominantly via a specific inhibition of tumour cell motility. This inhibitory effect of apigenin on tumour cell invasiveness in vitro demonstrates that apigenin may exert its anti-tumorigenic effect in vivo via inhibition of tumour cell penetration of the healthy tissue. PMID- 15523694 TI - Genotype phenotype correlations in Israeli colorectal cancer patients. AB - While genetic factors clearly play a key role in colorectal cancer (CRC) pathogenesis and in determining its phenotypic features, the precise genes that involved are largely unknown. To gain insight into these genes, consecutive Israeli CRC patients were genotyped using SNPs from within candidate genes: APC, beta-Catenin, K-RAS, DCC, P16, PTEN, RB1, P15, APOE, ERCC2, P53, MTHFR and hMSH2. Genotyping of consecutive, unselected colorectal cancer patients was done mostly by utilizing the MassARRAY technology (Sequenom) and to a lesser extent DGGE, ARMS and direct DNA sequencing. Correlation of genotypes with specific phenotypic features was carried out for all patients and separately for the Ashkenazim. Overall, 456 patients were analyzed, the majority (64.25%) being of Ashkenazi origin; mean age at diagnosis was 65.6 +/- 14 (range 25-90 years), and the mean follow-up was 4.7 +/- 0.28 (range 0-30 years). Statistically significant associations were noted between SNPs in beta-catenin and APOE and a positive family history of cancer (beta-catenin: p=0.034, APOE: p=0.033); tumor location and a DCC SNP (p=0.038) and the P53 R72P mutation and survival (p=0.0336). In Ashkenazi patients, ERCC2 and MTHFR genes' SNPs were associated with age at diagnosis (ERCC2: p=0.025, MTHFR: p=0.0005); a P53 polymorphism, APOE and Rb SNPs with a family history of cancer (P53 p=0.034;APOE p=0.04, Rb p= 0.022); DCC SNP with tumor location (p=0.014); and p15 SNP with tumor grade (p=0.032). This preliminary study shows that genetic factors play a role in determining CRC phenotypic features and that a larger cohort with longer follow-up is clearly needed. PMID- 15523695 TI - Matrilysin (MMP-7) promotes invasion of ovarian cancer cells by activation of progelatinase. AB - Although matrilysin (MMP-7) is overexpressed in various malignancies, few studies have evaluated its role in epithelial ovarian cancer (EOC) invasion and metastasis. We report that the secretion of MMP-7 in EOC is stimulated significantly by vascular endothelial growth factor (VEGF) and interlukin-8 (IL 8). We also examined the in vivo expression of MMP-7 in EOC and its effects on the in vitro invasion and progelatinase activation. We report that MMP-7 is overexpressed in ovarian cancer cell lines and EOC surgical specimens. DOV13 cells incubated with active rhMMP-7 significantly increased cellular invasion and proMMP-2 activation. RhMMP-7 also showed the ability to activate proMMP-2 and proMMP-9 in immortalized ovarian epithelial cell (IOSE-29) conditioned medium. In addition, rhMMP-7 was able to activate progelatinase in a concentration-dependent manner in vitro. TIMP-2 or the generic MMP inhibitor-GM6001 inhibited both the activation of proMMP-2 and the increased invasion of DOV13 cells promoted by rhMMP-7. By incubation of MMP2-TIMP-2 complex with equal molar rhMMP-7, MMP-2 was dissociated from the complex and activated in a time-dependent manner, suggesting that TIMP-2 helps to keep the latency of MMP-2. TIMP-2 also showed inhibitory effects on the MMP-7 induced increase of gelatinolytic activity in DOV13 and IOSE 29 conditioned media. A strong co-localization of MMP-7 and MMP-2 was observed in DOV13 cells and ovarian carcinoma permanent tissue sections. These results indicate MMP-7 is overexpressed in malignant ovarian epithelium and suggest MMP-7 may facilitate tumor cell invasion in vivo partly through the induction of progelatinase activation. PMID- 15523696 TI - Persistence of bone marrow micrometastases in patients receiving adjuvant therapy for breast cancer: results at 4 years. AB - We have previously developed a quantitative PCR (QPCR) technique for the detection of cytokeratin 19 (CK19) transcripts in blood and bone marrow and compared this to immunocytochemistry (ICC). Together, both have shown promise for monitoring therapeutic efficacy in patients with metastatic breast cancer. The aim of this study was to determine the feasibility and value of these assays for minimal residual disease (MRD) in monitoring efficacy of adjuvant therapy following surgery for primary breast cancer. Bone marrow aspirates and peripheral blood samples were taken at the time of surgery from patients with primary breast cancer and no evidence of metastases on conventional scans. These were tested for the presence of CK19 mRNA transcripts and cytokeratin positive cells. Follow-up bone marrow aspirates were taken at 3, 6, 12, 24, 36 and 48 months. Prior to surgery, 51% of patients displayed evidence of disseminated cancer cells in the bone marrow by either or both QPCR and ICC. Of 91 patients who had repeat samples assayed, 87% and 65% had positive results at some time using QPCR and ICC, respectively. All patients received adjuvant systemic therapy and in 44 cases where there was a positive result in either the pretreatment or 3-month aspirate, 32/44 (73%) showed a fall in CK19:ABL ratio (QPCR) and 15/24 (63%) showed a reduction in the number of cytokeratin-positive cells (ICC) during follow-up. These results indicate that MRD persists despite adjuvant therapy in a majority of patients with primary breast cancer up to 4 years following surgery. PMID- 15523697 TI - Renal cell carcinoma in relation to cigarette smoking: meta-analysis of 24 studies. AB - Renal cell carcinoma (RCC) accounts for 3% of adult deaths from cancer. The risk factors for its development are still under intense investigation. Although tobacco smoke is a risk factor, the data are inconsistent and the extent of the increased risk is unclear. Estimates from 19 case-control and 5 cohort studies were used. The case-control reports included 8,032 cases and 13,800 controls; the cohort estimates were based on 1,457,754 participants with 1,326 cases of RCC. The relative risk (RR) for RCC for ever smokers as compared to lifetime never smokers was 1.38 (95% confidence interval [CI] = 1.27-1.50). The RR for male smokers was 1.54 (95% CI = 1.42-1.68) and for female smokers was 1.22 (95% CI = 1.09-1.36). For men and women there was a strong dose-dependent increase in risk. Ever smoker men who had smoked 1-9, 10-20 or 21 or more cigarettes/day had a RR of 1.60 (95% CI = 1.21-2.12), 1.83 (95% CI = 1.30-2.57), or 2.03 (95% CI = 1.51 2.74), respectively. For women, the relative risks were 0.98 (95% CI = 0.71 1.35), 1.38 (95% CI = 0.90-2.11), or 1.58 (95% CI = 1.14-2.20), respectively. The advantages of smoking cessation were confirmed by a reduction in RR for those who had quit smoking for >10 years as compared to those who had quit for 1-10 years. Inhaled tobacco smoke is clearly implicated in the etiology of RCC, with a strong dose-dependent increase in risk associated with numbers of cigarettes smoked per day and a substantial reduction in risk for long-term former smokers. PMID- 15523698 TI - International uveal melanoma incidence trends in view of a decreasing proportion of morphological verification. AB - The introduction of eye-preserving therapies for uveal melanoma in the 1970s complicates time trend analyses of the uveal melanoma incidence because the proportion of morphologically verified uveal melanoma has been decreasing over the decades. We carried out incidence trend analyses, based on data from internationally accredited population-based cancer registries throughout the world that take missing data on topography, morphology and basis of diagnosis of eye tumours into account. We selected incidence data of cancer registries that were included in Cancer Incidence In 5 Continents, Volumes VI-VIII covering a registration period of at least 15 years (usually 1983 to 1997) and classified each eye cancer as morphologically verified uveal melanoma, clinically diagnosed uveal melanoma, uveal melanoma identified as DCO case (Death certificate only), possibly uveal melanoma, other eye tumour or unclassifiable eye tumour and calculated age-standardized incidence rates by 3-year calendar periods using the World Standard Population as the reference. The uveal melanoma incidence decline in the United States SEER Caucasian population is due mainly to an incidence decline in the early registration period (from 1974-76 to 1986-88). The data from France and Italy suggest a recent increase in incidence. Uveal melanoma diagnosed clinically increasingly contribute to the overall uveal melanoma incidence over time. Combining all registries, the proportion of morphologically verified uveal melanoma decreased from 82% in 1983-87 to 75% in 1993-97. Uveal melanoma incidence rates remained quite stable during the period 1983-97. The interpretation of uveal melanoma incidence trends is complicated by missing data on topography within the eye, morphology and basis of diagnosis. PMID- 15523699 TI - Downregulation of metastasis suppressor genes in malignant pheochromocytoma. AB - There is no reliable method currently available to predict malignant potential of pheochromocytoma based on conventional histology or genetic, molecular or immunohistochemical markers. Metastasis suppressor genes affect the spread of several cancers and, therefore, may provide promise as prognostic markers or therapeutic targets for malignant pheochromocytoma. We hypothesized that the downregulation of metastasis suppressor genes in malignant pheochromocytoma may play a role in malignant behavior. We applied quantitative real-time polymerase chain reaction (QRT-PCR) to 11 metastasis suppressor genes. These genes are known to be involved in the regulation of important cancer-related cellular events, such as cell growth regulation and apoptosis (nm23-H1, TIMP-1, TIMP-2, TIMP-3, TIMP-4, TXNIP and CRSP-3), cell-cell communication (BRMS-1), invasion (CRMP-1) and cell adhesion (E-Cad and KiSS1). The study included 15 benign and 10 malignant pheochromocytomas. Six metastasis suppressor genes (nm23-H1, TIMP-4, BRMS-1, TXNIP, CRSP-3 and E-Cad) were downregulated significantly in malignant compared to benign pheochromocytoma (p < 0.05, Mann-Whitney U-test). We applied a non-linear rule using median malignant value (MMV) as a threshold to use metastasis suppressor genes to distinguish malignant from benign samples. After cross-validation, the non-linear rule produced no errors in 10 malignant samples and 3 errors in the 15 benign samples, with an overall error rate of 12%. These results suggest that downregulation of metastasis suppressor genes reflect malignant pheochromocytoma with a high degree of sensitivity. Thus, we conclude that altered function of these metastasis suppressor gene pathways may play an important role in the malignant behavior of pheochromocytoma. PMID- 15523700 TI - Iron-fortified MRI: effects and applications of iron-induced NMR relaxation in biological tissues. PMID- 15523701 TI - Cytokeratin-positive cell dissemination in patients with in situ ductal carcinoma of the breast. PMID- 15523702 TI - Radon and other natural radionuclides in drinking water and risk of stomach cancer: a case-cohort study in Finland. AB - Very high concentrations of naturally occurring radionuclides are encountered in Finnish groundwaters and wells. Radon ingested through drinking water can cause considerable radiation to the stomach. We assessed the effect of natural uranium and other radionuclides in drinking water on the risk of stomach cancer. Subjects (n = 144,627) in the base cohort had lived outside the municipal tap water system during 1967-1980. A subcohort of 4,590 subjects was formed for use as a reference group by random sampling of the base cohort, with stratification by age and sex. Within the subcohort, 371 subjects had used drinking water from drilled wells prior to 1981. Stomach cancer cases within the subcohort were identified through a cancer registry, and cases using water from drilled wells were selected. Activity concentrations of radon, radium-226 and natural uranium in the drinking water were analyzed using radiochemical and alpha spectrometric methods. The median activity concentration of radon in well water was 130 Bq/l for both the 88 stomach cancer cases and the 274 subjects in the subcohort. Median radium concentrations were 0.007 Bq/l for cases and 0.010 Bq/l for the subcohort, with a median uranium concentration of 0.07 Bq/l for both groups. Risk of stomach cancer was not associated with exposure to radon or other radionuclides. The hazard ratio of stomach cancer was 0.68 for radon (95% CI 0.29-1.59 at 100 Bq/l water), 0.69 per Bq/1 for radium-226 (95% CI 0.33-1.47) and 0.76 per Bq/1 for uranium (95% CI 0.48-1.21). Our results do not indicate an increased risk of stomach cancer from ingestion of radon or other natural radionuclides through drinking water at these exposure levels. PMID- 15523703 TI - The interpoint distance distribution as a descriptor of point patterns, with an application to spatial disease clustering. AB - The topic of this paper is the distribution of the distance between two points distributed independently in space. We illustrate the use of this interpoint distance distribution to describe the characteristics of a set of points within some fixed region. The properties of its sample version, and thus the inference about this function, are discussed both in the discrete and in the continuous setting. We illustrate its use in the detection of spatial clustering by application to a well-known leukaemia data set, and report on the results of a simulation experiment designed to study the power characteristics of the methods within that study region and in an artificial homogenous setting. PMID- 15523704 TI - High-resolution MR imaging of mouse brain microvasculature using the relaxation rate shift index Q. AB - Magnetic resonance imaging (MRI) is a powerful method for in vivo quantification of tissue properties. It has been previously proposed that the index Q identical with Delta R2/(Delta R2*)2/3, where Delta R2 and Delta R2* are the spin echo and gradient echo relaxation rate shifts caused by the injection of an intravascular contrast agent, may be useful for characterizing microvasculature. In particular, Q is expected to correlate well with the density of microvessels. This study presents high-resolution in vivo Q-maps of normal mouse brain obtained with a superparamagnetic iron oxide contrast agent at a field of level of 9.4 T. Normative Q values are derived for several regions of interest and significant interregional variations are observed. Microvessel densities estimated from the Q maps are found to be in reasonable accord with histologically determined values. A possible application of Q-maps is the assessment of angiogenic activity in tumors. PMID- 15523705 TI - High-field magnetic resonance imaging of brain iron: birth of a biomarker? AB - The brain has an unusually high concentration of iron, which is distributed in an unusual pattern unlike that in any other organ. The physiological role of this iron and the reasons for this pattern of distribution are not yet understood. There is increasing evidence that several neurodegenerative diseases are associated with altered brain iron metabolism. Understanding these dysmetabolic conditions may provide important information for their diagnosis and treatment. For many years the iron distribution in the human brain could be studied effectively only under postmortem conditions. This situation was changed dramatically by the finding that T2-weighted MR imaging at high field strength (initially 1.5 T) appears to demonstrate the pattern of iron distribution in normal brains and that this imaging technique can detect changes in brain iron concentrations associated with disease states. Up to the present time this imaging capability has been utilized in many research applications but it has not yet been widely applied in the routine diagnosis and management of neurodegenerative disorders. However, recent advances in the basic science of brain iron metabolism, the clinical understanding of neurodegenerative diseases and in MRI technology, particularly in the availability of clinical scanners operating at the higher field strength of 3 T, suggest that iron-dependent MR imaging may soon provide biomarkers capable of characterizing the presence and progression of important neurological disorders. Such biomarkers may be of crucial assistance in the development and utilization of effective new therapies for Alzheimer's and Parkinson's diseases, multiple sclerosis and other iron related CNS disorders which are difficult to diagnose and treat. PMID- 15523706 TI - Joint modelling of bivariate longitudinal data with informative dropout and left censoring, with application to the evolution of CD4+ cell count and HIV RNA viral load in response to treatment of HIV infection. AB - Several methodological issues occur in the context of the longitudinal study of HIV markers evolution. Three of them are of particular importance: (i) correlation between CD4+ T lymphocytes (CD4+) and plasma HIV RNA; (ii) left censoring of HIV RNA due to a lower quantification limit; (iii) and potential informative dropout. We propose a likelihood inference for a parametric joint model including a bivariate linear mixed model for the two markers and a lognormal survival model for the time to drop out. We apply the model to data from patients starting antiretroviral treatment in the CASCADE collaboration where all of the three issues needed to be addressed. PMID- 15523707 TI - Repeated-measures models with constrained parameters for incomplete data in tumour xenograft experiments. AB - In cancer drug development, xenograft experiments (models) where mice are grafted with human cancer cells are used to elucidate the mechanism of action and/or to assess efficacy of a promising compound. Demonstrated activity in this model is an important step to bring a promising compound to humans. A key outcome variable in these experiments is tumour volumes measured over a period of time, while mice are treated with an anticancer agent following certain schedules. However, a mouse may die during the experiment or may be sacrificed when its tumour volume quadruples and then incomplete repeated measurements arise. The incompleteness or missingness is also caused by drastic tumour shrinkage (<0.01 cm3) or random truncation. In addition, if no treatment were given to the tumour-bearing mice, the tumours would keep growing until the mice die or are sacrificed. This intrinsic growth of tumour in the absence of treatment constrains the parameters in the regression and causes further difficulties in statistical analysis. We develop a maximum likelihood method based on the expectation/conditional maximization (ECM) algorithm to estimate the dose-response relationship while accounting for the informative censoring and the constraints of model parameters. A real xenograft study on a new anti-tumour agent temozolomide combined with irinotecan is analysed using the proposed method. PMID- 15523709 TI - Phosphine-promoted conversion of oxo(dithiolato)rhenium(v) into thio(thiolatoalkyl)rhenium(v) compounds. PMID- 15523708 TI - yDNA: a new geometry for size-expanded base pairs. PMID- 15523710 TI - Solid-phase synthesis of lipidated peptides. PMID- 15523711 TI - Analysis of poly(carbon suboxide) by small-angle X-ray scattering. PMID- 15523712 TI - Synthesis of beltenes by reactions of 5,6,11,12 tetradehydrodibenzo[a,e]cyclooctene with [CpCo(CO)2] derivatives. PMID- 15523713 TI - Hydrogen bond mediated enantioselectivity of radical reactions. PMID- 15523714 TI - Synthesis and photophysics of a neutral organic mixed-valence compound. PMID- 15523716 TI - Doxycycline- and tetracycline-regulated transcriptional silencer enhance the expression level and transactivating performance of rtTA. AB - BACKGROUND: The tetracycline-regulated transcriptional silencer (tTS) has been demonstrated to mitigate leaky expression of the tetracycline-inducible promoter under uninduced condition, and, when conjugated with reverse-type tetracycline controlled transactivator (rtTA), shows great promise for gene therapy. This effect was attributed to the effectiveness of tTS as a repressor of transcription at the tetracycline-regulated promoter. However, we observed an unexpected increase in transactivational activity by rtTA in the presence of tTS under inducible condition. METHODS: To explore the nature of this co-activational effect of tTS on rtTA, we examined the expression patterns of rtTA by Western blotting analysis of total cellular lysates or an enriched ubiquitinated pool of proteins under various conditions, including the one when proteasomal degradation is inhibited. RESULTS: We demonstrate tTS, in addition to its established role as a transcriptional silencer, can enhance rtTA expression level by salvaging rtTA from the ubiquitin-dependent proteasomal degradation pathway. Along with this finding, we also demonstrate that doxycycline, a commonly used tetracycline analogue, inhibits the susceptibility of rtTA to ubiquitin/proteasome-mediated degradation and enhances the expression level of rtTA. CONCLUSIONS: Taken together, our data establish an unappreciated role of doxycycline and tTS in tetracycline-regulated gene expression and the functionality of rtTA, and should shed light on the design of gene therapy vectors based on tetracycline-controlled transcriptional regulation systems. PMID- 15523717 TI - First-pass and equilibrium-MRA of the aortoiliac region with a superparamagnetic iron oxide blood pool MR contrast agent (SH U 555 C): results of a human pilot study. AB - The purpose of this study was to study different doses for first-pass and equilibrium phase MRA of aortoiliac vessels with a superparamagnetic iron oxide (SPIO) intravascular MR contrast agent (SH U 555 C) after single i.v. bolus injection. Sixteen healthy volunteers were prospectively enrolled into this single-blind, placebo-controlled clinical trial. SHU 555 C was injected as an i.v. bolus at stepwise increased dose levels of 5, 10, 20 and 40 micromol Fe/kg bodyweight (b.w.) corresponding to injection volumes of 0.01, 0.02, 0.04 and 0.08 ml/kg b.w. Serial high-resolution three-dimensional MRA of the aortoiliac vessels was acquired during first-pass and equilibrium, at 6 min intervals up to 42 min after contrast application using a breath-hold three-dimensional FLASH sequence on a 1.5 T scanner. Intravascular enhancement was calculated within the abdominal aorta and the inferior vena cava and a statistical analysis for significant differences in vessel enhancement was performed during the bolus and equilibrium phases. The visibility of vessels was ranked and effects of potential artifacts on image quality were graded for each time point and dose group. SH U 555 C showed a dose-dependent intravascular enhancement during the observation period (42 min). The highest dose of 40 micromol Fe/kg b.w. revealed the highest image quality during first-pass and equilibrium phases. The intravascular enhancement in the aorta increased dose-dependently from 5 to 40 micromol kg b.w. during first-pass and equilibrium phases (p<0.05). Intravascular signal inhomogeneities were observed at lower doses and decreased with increasing doses. First-pass MRA was diagnostic at doses of 10, 20 and 40 micromol Fe/kg b.w. For equilibrium MRA, a dose of 40 micromol Fe/kg b.w. was considered to be diagnostic. SH U 555 C proved to be a contrast agent with a high T1-effect suitable for both first-pass MRA comparable to gadolinium-enhanced MRA and high resolution equilibrium MRA up to 42 min post-injection (p.i.). PMID- 15523718 TI - Photocontrol of Smad2, a multiphosphorylated cell-signaling protein, through caging of activating phosphoserines. PMID- 15523719 TI - Self-assembly of cyclic metal-DNA nanostructures using ruthenium tris(bipyridine) branched oligonucleotides. PMID- 15523720 TI - Ru(II) tris(bipyridyl) complexes with six oligonucleotide arms as precursors for the generation of supramolecular assemblies. PMID- 15523722 TI - An aromatic anion receptor: anion-pi interactions do exist. PMID- 15523721 TI - The catalytic activity of "naked" gold particles. PMID- 15523723 TI - Nanofiber formation in the chemical polymerization of aniline: a mechanistic study. PMID- 15523724 TI - Total synthesis of apicularen A through transannular pyran formation. PMID- 15523725 TI - Unusual reactivity of a nickel N-heterocyclic carbene complex: tert-butyl group cleavage and silicone grease activation. PMID- 15523726 TI - Surface-confined metalloporphyrin oligomers. PMID- 15523727 TI - Arsenic-pi interactions stabilize a self-assembled As2L3 supramolecular complex. PMID- 15523730 TI - Nobel Prizes 2004: Protein degradation, olfactory system, strong interaction. PMID- 15523729 TI - Apology. Nanofibers from functionalized dendritic molecules. PMID- 15523731 TI - DNA-based barcodes, nanoparticles, and nanostructures for the ultrasensitive detection and quantification of proteins. PMID- 15523732 TI - Helix inside a Helix: encapsulation of hydrogen-bonded water molecules in a staircase coordination polymer. PMID- 15523733 TI - Building molecular minerals: all ferric pieces of molecular magnetite. PMID- 15523734 TI - Self-assembly of interpenetrating coordination nets formed from interpenetrating cationic and anionic three-dimensional diamondoid cluster coordination polymers. PMID- 15523735 TI - Modulation of compactness and long-range interactions of unfolded lysozyme by single point mutations. PMID- 15523736 TI - Synthesis of a large-scale highly ordered porous carbon film by self-assembly of block copolymers. PMID- 15523737 TI - Asymmetric double proton transfer of excited 1:1 7-azaindole/alcohol complexes with anomalously large and temperature-independent kinetic isotope effects. PMID- 15523739 TI - Abstracts of the ISHR Satellite Meeting: Cellular Injury in Ischemia. Kruger Park, South Africa, 13-16 August 2004. PMID- 15523738 TI - Endocrine-related resources from the National Institutes of Health. PMID- 15523740 TI - Abstracts of the American Association for Cancer Education 38th Annual Meeting. October 14-17, 2004, Baltimore, Maryland, USA. PMID- 15523741 TI - Abstracts of the 8th Annual Scientific Meeting of the Japanese Heart Failure Society. October 2, 2004, Gifu, Japan. PMID- 15523742 TI - Bibliography. Current world literature. Gastrointestinal infections. PMID- 15523743 TI - Management of chronic asthma. PMID- 15523744 TI - Criteria for determining disability in speech-language disorders. PMID- 15523745 TI - Utility of blood pressure monitoring outside of the clinic setting. PMID- 15523746 TI - Management of neonatal hyperbilirubinemia. PMID- 15523747 TI - Criteria for determining disability in infants and children: low birth weight. PMID- 15523748 TI - Effect of the supplemental use of antioxidants vitamin C, vitamin E, and coenzyme Q10 for the prevention and treatment of cancer. PMID- 15523749 TI - Hyperbaric oxygen therapy for brain injury, cerebral palsy, and stroke. PMID- 15523750 TI - Intravenous administration of N-acetylcysteine: interference with coagulopathy testing. PMID- 15523751 TI - What is the rate of adverse events with intravenous versus oral N-acetylcysteine in pediatric patients? PMID- 15523752 TI - Is capnography necessary for propofol sedation? PMID- 15523753 TI - Ecstasy and acute myocardial infarction. PMID- 15523754 TI - Get the lead out. PMID- 15523756 TI - Education of veterinarians in Europe: the basis for recent change. PMID- 15523755 TI - Oral hypoglycemics sold as Valium on the streets: a case report. PMID- 15523757 TI - Replacement of an aortic Starr-Edwards ball valve prosthesis 28 years after implantation. PMID- 15523758 TI - Just put me to sleep...please! Ethical issues in palliative and "terminal" sedation. PMID- 15523759 TI - Terminal sedation: a Jewish perspective. PMID- 15523760 TI - Terminal sedation: a Catholic perspective. PMID- 15523761 TI - Personal and professional integrity in clinical medicine. PMID- 15523762 TI - Eukaryotes: not beyond compare. PMID- 15523763 TI - AIDS and tuberculosis - a lethal combination. PMID- 15523764 TI - Recent references. PMID- 15523765 TI - Long-term care and Medicaid. PMID- 15523766 TI - Biomonitoring. PMID- 15523767 TI - Autism Disorders. PMID- 15523768 TI - Two campaigns, two health plans. PMID- 15523769 TI - Stark II final regulations: hot issues for physician practices. AB - The Stark law, first passed in 1989 and repeatedly expanded and clarified since then, has long created questions for group practices. Its primary intent is to prohibit physician referrals to entities in which the physician has a financial interest, but the details have proven confusing. The Stark law's Phase II regulations, as put forth by the Centers for Medicare and Medicaid Services, went into effect in July and attempt to offer further clarification. In this article the authors explain some of the complexities of this most recent set of Stark law regulations, especially as they apply to smaller, multi-specialty practices. PMID- 15523770 TI - Procedural tort reform in the United States: lessons from other nations. Part I : The background. AB - As currently constituted, the tort system in the United States has induced distortions that have resulted in a crisis in medical litigation with escalating awards, as well as in product liability. This article outlines views of the structural defects in the American system and contrasts it with the experience in Britain and the major commonwealth nations. In particular, the author stresses the inefficiencies of the civil jury system. In a subsequent article, he will offer some possible remedies to this problem. PMID- 15523771 TI - Special considerations for hiring an associate. AB - If you 're thinking about hiring an associate for your medical practice, you will have many things to consider and many decisions to make. This article weighs the pros and cons of associateships and provides a 12-question quiz to help you determine whether an associate will be a good fit for your goal, philosophy, needs, and personality. It suggests helpful tips for recruiting and interviewing top-notch applicants to your associateship, including effective interview questions. It also offers advice about evaluating associateship applicants and suggests typical compensation arrangements. This article also includes questions to answer in your associateship contract and practical advice about restrictive covenants and externships. Finally, the article offers 12 suggestions for getting your new associate up to speed right away in your practice. PMID- 15523772 TI - Disaster recovery for electronic data: are you prepared? AB - Disasters that affect medical data may come in many forms, but those that can harm sensitive computer information are not necessarily dramatic. Even so, the problems that can arise if data is lost can be severe. This article discusses the issues of how to recover data once an information technology disaster has occurred and how to prevent or guard against disaster. Not all disasters are preventable, but many are. Even if a disaster is not preventable, certain practices in advance can ease the process of recovery. PMID- 15523773 TI - Computer briefs: Internet telephone service. PMID- 15523774 TI - Implementing patient satisfaction survey findings into a customer service action plan. AB - Patient satisfaction surveys have become popular gauges of practice efficiency and are among the markers used by third-party payers to measure health-care quality. Although surveys may yield valuable information for providers to improve their services, these results most often are assigned a low priority and not applied in actual practice. This article briefly outlines the basic features of a patient satisfaction survey and details specific steps that managers may follow to implement their findings. PMID- 15523775 TI - Return of premium term life: is it for real? AB - Physicians who feel they are wasting their money in paying premiums for a typical 30-year-level term life insurance policy may find help in a return-of-premium term life insurance policy. Premiums are higher, but all the money is refunded at the end of the term if the policyholder has not died. Estimates are that 93 percent of policyholders outlive the term of their life insurance. PMID- 15523776 TI - Coding for split visits for Medicare patients: preventive medicine and office visit on the same day. PMID- 15523777 TI - Hospitals expect capital expenditures to increase. PMID- 15523778 TI - Managing medical groups: 21st century challenges and the impact of physician leadership styles. AB - Physician group managers and administrators charged with leading medical groups in the 21st century face a set of old and new challenges and opportunities. Leadership is assumed to make the difference between a successful and not-so successful medical group. Yet, there is little research about how physician manager leadership styles contribute to the success of medical group practices. This article is a study of physician leadership styles using the DiSC, based upon a sample of 232 physician managers. Dominance (D) and conscientiousness (C) were the two dominant styles found in this study. Moreover, the two dominant combination leadership styles fall under the categories of the "creative" and the 'perfectionist." The article formulates practical recommendations for both physician managers and administrators for leading medical groups to respond more effectively to the challenges and opportunities facing medical groups in the 21st century. PMID- 15523779 TI - Bush administration's proposed solutions for controlling health-care costs. PMID- 15523780 TI - Common misconceptions among surgeons named in malpractice cases. PMID- 15523781 TI - The (mis)application of management science in medicine: a flawed concept. AB - The business of medicine once again finds itself in the throes of rapidly escalating costs, concerns about quality of care, and demands for efficiency while simultaneously enhancing quality. Considerable effort has already been spent in trying to improve costs, quality, and patient satisfaction. The apparent failure to do so may be the result of a fundamental misunderstanding of the salient features of clinical practice and the misapplication of quality improvement techniques. This article explores some of the significant issues and offers potential new directions. PMID- 15523782 TI - Consumer price index trends in 2004. PMID- 15523783 TI - Stormwater. PMID- 15523785 TI - Oxidation-reduction potential changes in aeration tanks and microprofiles of activated sludge floc in medium- and low-strength wastewaters. AB - Real-time control of aeration tank operation is key to high-efficiency pollutant removal and energy savings. One of the aims of this study was to examine the potential for using redox potential (oxidation-reduction potential [ORP]) to indicate wastewater quality online in aeration tanks treating medium (chemical oxygen demand [COD] of 70 to 150 mg/L) and low (COD of 15 to 30 mg/L) pollutant concentration wastewaters. The field-scale data provide a good relationship between ORP values and nutrient removal along the length of the aeration tanks. The ORP values increased dramatically as organic matter was removed along the aeration tanks, indicating the improvement of the bulk liquor redox status. Dissolved oxygen higher than 1.0 mg/L was necessary for good biodegradation and improvement of the liquid redox status. Nitrification occurred at higher ORP values (380 to 420 mV) than was the case for organic substrate oxidation (250 to 300 mV). The microprofiles obtained from microelectrode measurements substantiate the heterogeneity of the microbial processes inside activated sludge flocs. Because of microbial oxygen utilization, the aerobic region in the activated sludge floc was limited to the top layer (0.1 to 0.2 mm) of the activated sludge aggregate present in medium-strength wastewater, with an anoxic zone dominating inside the flocs. When dissolved oxygen in the bulk water was higher than 4.0 mg/L, the anoxic zone inside the floc disappeared. At low wastewater pollutant concentrations, the ORP and dissolved oxygen inside the activated sludge aggregates were higher than those from medium-strength wastewater. The prospect of using ORP as an online control approach for aeration tank operation and the potential reasons for activated sludge floc size varying with pollutant strengths are also discussed. PMID- 15523784 TI - Denitrification of nitrate wastewater using packed-bed columns. AB - Columnar packed-bed (PB) reactors with a specific surface area of 127 m2/m3 were investigated in this study for treating nitrate wastewater. This study demonstrated that a single-stage packed bed was able to achieve total nitrogen (TN) and chemical oxygen demand (COD) removal efficiencies higher than 83 and 75%, respectively. The highest achievable TN and COD removal rates were 47.2 g N/m2 x d and 158.0 g COD/m2 x d, respectively. The substrate removal rate in the PB column was found to follow half-order reaction kinetics, with a reaction coefficient, kappa, of 53.62 (mg/L)1/2/d. A dual-stage PB system was capable of achieving TN and COD removal efficiencies greater than 99 and 98%, respectively. Effluent TN and COD concentrations less than 6.5 mg NO3(-)-N/L and 50.0 mg COD/L, respectively, were obtained when the dual PB system was used. PMID- 15523786 TI - Carbon and nutrient removal from on-site wastewater using extended-aeration activated sludge and ion exchange. AB - The need to improve on-site wastewater treatment processes is being realized as populations move into more environmentally sensitive regions and regulators adopt the total maximum daily load approach to watershed management. Under many conditions, septic systems do not provide adequate treatment; therefore, advanced systems are required. These systems must remove significant amounts of biochemical oxygen demand (BOD) and suspended solids, and substantially nitrify, denitrify, and remove phosphorus. Many existing advanced on-site wastewater systems effectively remove BOD, suspended solids, and ammonia, but few substantially denitrify and uptake phosphorus. The purpose of this research was to design and test modifications to an existing on-site wastewater treatment system to improve denitrification and phosphorus removal. The Nayadic (Consolidated Treatment Systems, Inc., Franklin, Ohio), an established, commercially available, extended-aeration, activated sludge process, was used to represent a typical existing system. Several modifications were considered based on a literature review, and the option with the best potential was tested. To improve denitrification, a supplemental treatment tank was installed before the Nayadic and a combination flow splitter, sump, and pump box with a recirculation system was installed after it. A recirculation pump returned a high proportion of the system effluent back to the supplemental treatment tank. Two supplemental treatment tank sizes, three flowrates, and three recirculation rates were tested. Actual wastewater was dosed as brief slugs to the system in accordance with a set schedule. Several ion-exchange resins housed in a contact column were tested on the effluent for their potential to remove phosphorus. Low effluent levels of five-day biochemical oxygen demand, suspended solids, and total nitrogen were achieved and substantial phosphorous removal was also achieved using a 3780-L supplemental treatment tank, a recirculation ratio of 5:1, and a fine-grain activated aluminum-oxide-exchange media. Good results were also obtained with an 1890-L supplemental treatment tank and a recirculation ratio of 3:1. The most significant benefit of the supplemental treatment tank, in combination with the recirculation system, appears to be the low nitrogen concentration dosed to the Nayadic. By reducing the nitrogen concentration and spreading out its mass over time during no-flow periods, the Nayadic's inherent low-level denitrifying capacity was more closely matched and effective treatment was achieved. PMID- 15523787 TI - Monitoring of bacterial morphology for controlling filamentous overgrowth in an ultracompact biofilm reactor. AB - This research was part of a study of filamentous growth and control in an ultracompact biofilm reactor (UCBR). Morphologies of biofilm and filamentous bacteria in the UCBR were investigated. Ethanol was used as a substrate and sodium hypochlorite was applied as a toxicant to control filamentous growth. The results indicated that factors such as chemical oxygen demand, surface loading rate, pH, and dissolved oxygen could initiate filamentous overgrowth in the UCBR. Different biofilm and filamentous morphologies in the UCBR were observed under different operational conditions. Chlorination was an effective approach to control filamentous growth during and after biofilm formation. Proper chlorine dosing had no effect on biofilm, but killed filaments. Overdose of chlorine damaged biofilm and caused adverse effects such as low treatment efficiency, media clogging and washout, and biofilm color change in the reactor. Frequent monitoring of the morphologies of filaments and biofilm was needed during chlorination to prevent chlorine overdose. PMID- 15523788 TI - A review of biofouling and its control in membrane separation bioreactors. AB - Membrane separation technology is increasingly becoming an important innovation in biological wastewater treatment. Biofouling of the membrane is a major factor affecting the efficient and economic operation of membrane separation bioreactors (MBRs). This review summarizes the state-of-the-art progress in understanding the mechanisms and factors affecting membrane biofouling and the strategies for biofouling control. Biofouling mechanisms include the adsorption of soluble and suspended extracellular polymers on membrane surfaces and in membrane pores, the clogging of membrane pore structure by fine colloidal particles and cell debris, and the adhesion and deposition of sludge cake on membrane surfaces. Design and operating conditions of membrane modules and materials, hydrodynamic conditions in MBRs, process and environmental conditions of activated sludge systems, and the physicochemical properties of the wastewater are the dominant factors determining membrane biofouling. Current strategies to control biofouling include periodic relaxation, backwashing, chemical cleaning, and possible manipulation of hydrodynamic conditions and sludge properties. Achieving full integration of MBRs in wastewater treatment technology requires further research and development. Fundamental information on the bacteria, colloid, and membrane interaction, developed through multimethod and multiscale approaches, is particularly needed. PMID- 15523789 TI - Removal of struvite to prevent problems associated with its accumulation in wastewater treatment works. AB - Deposition of struvite within the infrastructure of wastewater treatment works can cause significant operating difficulties. A laboratory-scale, two-stage, continuously stirred tank reactor was developed to remove struvite to prevent problems associated with its deposition. Before any experimental work was conducted, the mixing regime within the system was determined using a pH-tracer response method. Perfect mixing conditions were achieved once the stirring speed reached 500 rpm. Experiments were conducted examining the effects of various process parameters such as retention time, pH, recycling rate, and ammonium concentration. By increasing the hydraulic retention time, the residual magnesium concentration decreased, with the optimum conditions occurring at 125 minutes. PMID- 15523790 TI - Use of extant kinetic parameters to predict effluent concentrations of specific organic compounds at full-scale facilities. AB - To use the results of kinetic tests to predict effluent concentrations of specific contaminants in activated sludge systems, the fraction of the biomass that has an ability to degrade the test compound (i.e., competent biomass) must be estimated. A calibration procedure was developed to assess the competent biomass concentration because the chemical oxygen demand (COD) fraction tended to underestimate the degrading fraction for three of the four test compounds. Acetone, for instance, had a measured influent COD fraction of 0.08%, and the actual competent fraction was estimated to be 2.3%, based on the model calibration. Once the competent biomass fraction in the mixed liquor was determined, the extant kinetic parameters were subsequently used to predict activated sludge system performance. Predicted effluent concentrations were within 2, 5, and 16% of the average measured concentrations for acetone, linear alkylbenzene sulfonate, and furfural, respectively. Day-to-day predictions for these compounds were less accurate, possibly because of the non-steady-state nature of the activated sludge systems studied. The difference between the fraction of the influent COD contributed by the target compounds and the competent biomass fraction in the mixed liquor was found to be more significant when the target compound contributed less than 1% of the influent organic matter. The chemical structure of the target compound and chemical composition of the influent likely had an effect on the resulting competent biomass concentration. The total maximum growth rate, microX, was observed to be independent of the influent concentration of acetone and furfural, thus suggesting that the competent biomass concentration for these compounds was not affected by the changes in their influent concentrations. Consequently, a majority of competent biomass growth resulted from the degradation of other substrates, resulting in a competent biomass concentration significantly higher than predicted based on the influent COD fraction contributed by the test compound. PMID- 15523791 TI - Aerobic sludge digestion under low dissolved oxygen concentrations. AB - Low dissolved oxygen (DO) concentrations occur commonly in aerobic digesters treating thickened sludge, with benefits of smaller digester size, much reduced aeration cost, and higher digestion temperature (especially important for plants in colder areas). The effects of low DO concentrations on digestion kinetics were studied using the sludge from municipal wastewater treatment plants in Akron, Ohio, and Los Lunas, New Mexico. The experiments were conducted in both batch digestion and a mixed mode of continuous, fed-batch, and batch operations. The low DO condition was clearly advantageous in eliminating the need for pH control because of the simultaneous occurrence of nitrification and denitrification. However, when compared with fully aerobic (high DO) systems under constant pH control (rare in full-scale plants), low DO concentrations and a higher solids loading had a negative effect on the specific volatile solids (VS) digestion kinetics. Nonetheless, the overall (volumetric) digestion performance depends not only on the specific digestion kinetics, but also the solids concentration, pH, and digester temperature. All of the latter factors favor the low DO digestion of thickened sludge. The significant effect of temperature on low DO digestion was confirmed in the mixed-mode study with the Akron sludge. When compared with the well-known empirical correlation between VS reduction and the product (temperature x solids retention time), the experimental data followed the same trend, but were lower than the correlation predictions. The latter was attributed to the lower digestible VS in the Akron sludge, the slower digestion at low DO concentrations, or both. Through model simulation, the first-order decay constant (kd) was estimated as 0.004 h(-1) in the mixed-mode operations, much lower than those (0.011 to 0.029 h(-1)) obtained in batch digestion. The findings suggested that the interactions among sludges with different treatment ages may have a substantially negative effect on digestion kinetics. The use of multistage digesters, especially with small front-end reactors, may be advantageous in both "process" kinetics and "biological reaction" kinetics for sludge digestion. PMID- 15523792 TI - Analysis of microbiological characteristics of wastewater along the polishing sequence of a treatment plant. AB - Microbial populations were studied by plate counts and flow cytometry along the polishing sequence of a wastewater plant. The comparison between plate count and flow cytometry showed comparable trends, but plate counts detected less than 10% of the total intact bacteria counted by flow cytometry. Six months monitoring showed that, in spite of the high ozone doses, the disinfection effect of ozonation was insignificant. This is in agreement with the variability of ozone demand, not only related to chemical oxygen demand or total suspended solids. Membrane-intact bacterial population decreased after ozonation, slightly regrew between ozone and biological activated carbon (BAC), and sharply increased after BAC, probably for damaged bacteria regrowth and saprophytes release; BAC effluent had the highest bacterial counts. Preliminary investigations on the effluent microbial composition showed that the beta Proteobacteria subclass is the most represented in the BAC effluent, whereas the alpha subclass is the most sensitive to ozone effect. PMID- 15523793 TI - Heterotrophic community-level physiological profiles of domestic wastewater following treatment by small constructed subsurface flow wetlands. AB - Seasonal treatment performance of small-surface flow wetlands was evaluated during their second operational year and compared with community-level physiological profiles (CLPP) of the heterotrophic bacterial community obtained from sole-carbon source utilization patterns in BIOLOG GN (Haywood, California) microplates. The CLPP patterns varied significantly by season, indicating reduced functional diversity in the heterotrophic community during warmer months of active plant growth (April through October). Principal component analysis (PCA) suggested that the functional community differed in planted versus unplanted wetlands during the growing season. Wetlands generally improved wastewater quality; however, treatment performance was reduced in the second year. Despite differences in the heterotrophic community suggested by CLPP, treatment efficiency with respect to removal of five-day biochemical oxygen demand or reduction in fecal indicator organisms generally was not significantly changed as a function of growing season or plant treatment. PMID- 15523794 TI - Evaluation of high-rate clarification for wet-weather-only treatment facilities. AB - High-rate clarification (HRC) processes are well suited for enhanced primary treatment of wet-weather flows in combined sewer systems. Suspended solids removal in excess of 75% of influent concentrations can be achieved consistently. Chemical oxygen demand and five-day biochemical oxygen demand removal are better than 60%. However, although optimal treatment could be achieved in a matter of minutes when the units were started full, a delay of up to one-half hour was observed when the units were started empty. Operational strategies that minimize this effect need to be developed in full-scale tests before HRC processes can be implemented with confidence in wet-weather-only facilities. PMID- 15523795 TI - Brain teaser: how to find a cure for diagnostics' ills. PMID- 15523796 TI - Tories give choice a blue rinse and spin. PMID- 15523797 TI - Schools ideas. Something to shout about. PMID- 15523798 TI - Data briefing. Waiting times. PMID- 15523799 TI - Clinical management. Where medicine meets management. Those who scan. AB - Because of a national shortage of radiologists, sonographers in one trust are performing 95 per cent of ultrasound scans. Using a team approach and flexible working, waiting lists have gone down from 20-24 weeks to four to six weeks for most procedures. The model could be applied to other radiology departments. PMID- 15523800 TI - Mental health. The plus side of zero stars. AB - A zero-starred mental health trust is battling to turn around its reputation. The trust believes it is reversing its history of vexed relations with other trusts and agencies. The trust is expecting a pounds sterling 29m boost for accommodation. PMID- 15523801 TI - Finance. Happy together. PMID- 15523802 TI - HSJ people: regional focus on the North West. Talkin' about regeneration. PMID- 15523803 TI - [Special clinical manifestations of Helicobacter pylori infection in children and adolescents]. AB - During the period 1989-1996, we retrospectively investigated 104 children and adolescents with Helicobacter pylori infection diagnosed by gastroscopy. The median age was 12.11 +/- 3.31 years, 55% were female and 45% male. The chief complaint of 92% of the children was abdominal pain, mainly epigastric. The typical macroscopic finding was nodular gastritis. The most common microscopic picture was chronic superficial gastritis with lymphatic follicles and the presence of Helicobacter pylori. We demonstrated correlation between these gastroscopic and histological findings. Children who suffered from peptic ulcer were older than those with nodular gastritis. Most of the Helicobacter pylori positive patients were treated with triple drug therapy. The combination of drugs changed throughout the years according to the newly accepted strategies in the treatment of Helicobacter pylori. The majority of children reported a decrease in symptoms on completion of therapy. Urea breath test was an efficient and convenient method for the follow-up of patients with Helicobacter pylori infection. PMID- 15523804 TI - [Comparison between D-dimer levels and lung scan in patients with suspected pulmonary embolism]. AB - Pulmonary embolism (PE) is a critical situation that requires prompt diagnosis and treatment. Lately, the use of D-Dimer (DD) blood level test was recommended when PE is suspected. The goal of this work was to evaluate the association between DD levels and the results of lung scan in patients with suspected PE. 84 patients who were admitted due to suspected PE and met the inclusion criteria of having both DD test and lung scan within less than 7 days apart, were included in our study (age = 76 +/- 14, females = 58, males = 26). Comparison of lung scan results, DD level and diagnosis of PE at discharge was carried out. The subjects were classified into 2 groups according to the DD levels: group A = DD > or = 250 ng/mL--abnormal test, group B = DD < 250 ng/mL--normal test. Group A includes 67 patients, of which 29 (43%) were diagnosed as PE. In these subgroup lung scan showed high Probability (HP) for PE in 19 (66%), medium probability (MP) in 9 (31%) and low probability (LP) in 1 (3%). From the 38 (57%) patients who were not diagnosed with PE (non-PE), 5 (13%) had HP lung scan, 4 (11%) MP and 29 (76%) LP. Group B includes 17 patients, in which only one (6%) had diagnosed with PE and the rest (n = 16, 94%) had no PE. Of the latter subgroup 15 (94%) had LP lung scan and one MP scan. The one patient who was diagnosed with PE had a HP lung scan that was also confirmed with CT-angiography. DD and lung scan had similar sensitivity (96.7%) and high negative predictive value (NPV = 94.1% and 97.8% accordingly) for PE. On the other hand DD specificity is much lower (29.6%) in comparison to lung scan (81.5%). The results of this work support the use of DD test prior to lung scan, in order to rule out PE with relatively high probability. PMID- 15523805 TI - [Thyroid surgery without drains]. AB - Wound drainage after thyroidectomy was considered to be a routine procedure. However, it has recently been noted that there is a rising trend in the literature to limit wound drainage to selected cases, or even obviate the need for drains altogether. This article presents 100 cases of patients undergoing hemi or total thyroidectomy without drains over a 5 year period. Only two cases of hemorrhage were observed and the overall hospital stay was shortened. Drains after thyroidectomy can be avoided in most cases and should be used sparingly when indicated. PMID- 15523806 TI - [Splenic infarction as an unusual cause of left pleural effusion]. AB - Many etiologies lead to pleural effusion. The pathogenetic cause is usually located either in the lung parenchyma or in the pleura. Subphrenic causes that lead to pleural effusion are uncommon. Several reports elaborated on the role of splenic hemorrhages in the genesis of left-sided pleural effusion. Splenic infarction is a rare etiology of left-sided pleural effusion, and it has rarely been described in medical literature. We present a case study of an elderly female patient who suffered from polycythemia vera for more than a decade, and was hospitalized for left-sided pleural effusion that appeared following left upper abdominal pain. PMID- 15523807 TI - [Development of a Hebrew questionnaire to be used in epidemiological studies to assess physical fitness--validation against sub maximal stress test and predicted VO2max]. AB - BACKGROUND: The physical activity questionnaire is typically chosen for population studies because it is practical and does not interfere with the behavior of the surveyed individual. This instrument can be adapted for the particular studied population, and in terms of accuracy it is both reliable and valid. AIM: To develop a valid and appropriate measure to assess physical fitness according to the level of physical activity throughout the day, and to estimate the relation between a subjective questionnaire measurement evaluating physical activity, to predicted VO2max, as an objective measure of physical fitness. METHODS: Three hundred healthy subjects, 150 men and 150 women, aged 25-65 years old participated in this study. All subjects were routinely examined in the Sheba Medical Center Executive Screening Survey. They were asked to complete a self administered questionnaire and report on the level of physical activity at work, the level of sports activity and the level of leisure time non-sport activity. The index calculated from the questionnaire was compared to the result from a sub maximal exercise test, which they performed in The Executive Screening Survey, to composite a valid questionnaire to estimate physical fitness. RESULTS: A high and statistically significant correlation (r = 0.64, P < 0.01) was found between the indexes of physical activity calculated from the questionnaire to the predicted VO2max, measured from the sub-maximal exercise test. The reliability of the questionnaire was examined by test-retest and a high and statistically significant correlation (r = 0.89, P < 0.01) was found between the first and the proceeding one month administration of the questionnaire. The final modal to predict VO2max included a total index of physical activity, gender, BMI, age and resting heart rate. Overall this model predicted 58% of the VO2max. CONCLUSIONS: The self-administered questionnaire developed in this study has high construct validity and a significant correlation to predicted VO2max, with high sensitivity in both sexes and among active and non-active subjects. It is therefore an appropriate tool to be used in epidemiological studies. PMID- 15523808 TI - [Relatives' satisfaction with the care of elderly patients in skilled nursing wards]. AB - BACKGROUND: Skilled Nursing Wards (SNW) in Israel are purposely equipped and staffed for treatment of patients that, in addition to functional dependence, suffer from active diseases which require close medical supervision. Patients' surveys on satisfaction with health care are increasingly accepted as an element of quality assessment. However, given the high rate of communication disorders present in these patients, this method is not feasible for use in SNWs. OBJECTIVE: We initiated a survey of patients' relatives--based on a questionnaire specifically designed to reflect their satisfaction with the level of care provided. METHODS: Respondents were asked to rank their level of satisfaction with medical, nursing and structural aspects of the care in the SNW on a 1-5 Likert Scale. RESULTS: Relatives of 147 SNW residents answered the questionnaire. The overall level of satisfaction was high, all components being ranked at 4.2 or above. Four point fifty-five was the average rating given to the medical component of the care, 4.39 to the nursing aspect and 4.32 to the structural section. Five points were awarded to the medical component by 60% of the participants, to the nursing element by 55% of respondents and to the structural section by 40% of respondents. No differences were observed between the rankings of frequent and less frequent visitors. All respondents were highly cooperative and expressed their appreciation of the initiative to perform a satisfaction survey. CONCLUSION: The results show that the families of the skilled nursing patients that participated in this study are satisfied with the health care provided to their relatives. Such surveys contribute to the quality of care and should be considered for routine/periodical use in long-term care settings. PMID- 15523809 TI - [Vaccines for all occasions]. AB - The strategy of immunizing a population at risk of infectious disease has been enormously successful medically and has also proven to be cost effective. Development of effective immunogens, that induce active immunization, is a long process that requires careful monitoring and assurance of short and long term safety, induction of protective immunity and proven efficacy in preventing the disease. A successful immunization program is also dependent on delivery of the vaccine to as many susceptible individuals as possible, so as to attain herd immunity. Passive immunization with antibodies, usually used prior to the development of active vaccines has also been remarkably effective. The special circumstances of the field and crowded conditions have demanded that the Medical Corps of the Israeli army cater for the needs of our soldiers. In this issue, the past achievements and current immunization policy are outlined for the first time. Their contribution to the health of our soldiers is commendable. Close monitoring of the epidemiology of infectious disease in the special circumstances of field conditions has prompted successful programs to markedly reduce infectious hepatitis A by passive immunization with gamma globulin in the past and, nowadays, with the killed active viral vaccine. In addition, prevention of influenza by killed viral vaccine and invasive bacterial disease by Neisseria meningitidis with multivalent polysaccharide vaccines are being used. This group has also improved hygienic conditions in the field to cope with shigellosis and salmonella infections. Research in the development of effective vaccines for protection of shigellosis has also been addressed by this group. New challenges posed by the emerging infectious diseases and the possible effects of bioterrorism are certain to keep this group on their toes. PMID- 15523810 TI - [Patients' and relatives' satisfaction with the care in geriatric wards]. AB - The geriatric wards in Israel are well equipped and staffed in order to treat elderly patients who need rehabilitation or special nursing and special medical supervision for active diseases. Health institutions and hospitals in Israel and in the modern world strive to satisfy both patients and relatives. Patients', relatives' and caregivers' satisfaction are important quality measurements of the medical and nursing treatment as well as the level of service provided. Patients' surveys of satisfaction with health care are increasingly accepted as an element of quality assessment and even as a management tool for changing and improving nursing and medical care, quality of service, physical and environmental conditions and hygiene of health institutions. Satisfaction surveys should be performed anonymously at least once a year. The quality of service evaluation and publication is in itself a stimulation to motivate the continuous improvement of the medical and non-medical staff. PMID- 15523811 TI - [Aromatase inhibitors--new applications in gynecology]. AB - P-450 aromatase inhibitors, designed for suppressing estradiol production, were first approved for the treatment of advanced breast cancer. Recent studies have provided evidence that aromatase inhibitors may be effective in the short term for induction of ovulation and in the long-term for treatment of endometriosis. Based on current data, the role of aromatase inhibitors in the management of various gynecological conditions may soon be widely determined. PMID- 15523812 TI - [Omega-3 fatty acids, fish, fish oil and cardiovascular disease--a review with implications to Israeli nutritional guidelines]. AB - Evidence from epidemiological and randomized controlled trials shows beneficial effects of omega-3 (n-3) fatty acids from fish and plant sources on cardiovascular disease (CVD), especially in patients with preexisting CVD. The optimal dose of n-3 is not yet determined, but prospective secondary prevention studies suggest that the addition of 0.5-1.8 grams/day of marine-derived eicosapentaenoic acid and docosahexaenoic acid, or plant derived alpha-linolenic acid at a dose of 1.5-3 grams/day significantly reduce subsequent cardiac events and mortality. These data have led the American Heart Association Dietary Guidelines committee to recommend to the general population the consumption of at least two servings of fatty fish per week, in addition to vegetable oils high in alpha-linolenic acid. The risk of adverse effects and toxicity from contaminants at this dose is low. The amount of daily n-3 fatty acids recommended for patients with coronary heart disease is 1 gram/day. In patients who cannot consume this dose of n-3 fatty acids through diet alone, addition of n-3 supplements should be considered. Higher doses of contaminant-free n-3 supplements, 2-4 grams/day, can be used in the treatment of hypertriglyceridemia. Data on the content of n-3 fatty acids and contaminants in Israeli bred fish is limited. Thus, caution should be exercised when applying these recommendations to the Israeli fish market. PMID- 15523813 TI - [Weighing the efficacy and safety of herbal medicine: the case of Kava-Kava]. AB - Kava-Kava is an indigenous plant in the Pacific islands and has been traditionally used for centuries for both ceremonial and social purposes. The efficacy of the plant for treatment of anxiety states was recently acknowledged in randomized controlled trials. Although these trials support the safety of Kava for short term usage, a major concern was raised in accordance with emerging reports that relate Kava use to major hepatic damage, including a few cases of fulminant hepatitis that required liver transplantation or ended in death. As a result, most medical authorities in the West restricted or banned Kava use. This review discusses evidence of efficacy and safety in the use of Kava for treating patients with anxiety, as well as its meaning in the therapeutic context and patient-doctor dialogue. PMID- 15523814 TI - ["The best among the physicians to Gehinom (to hell)?"]. AB - BACKGROUND: At first glance, the well-known Mishnaic dictum, "The best among the physicians to Gehinom" (Kiddushin 4:14), is a harsh and sweeping denunciation of doctors and medicine. It is contrary to the respect and regard for physicians and medicine shown by the Talmud and Jewish tradition across the generations. AIM: This article explains the statement in its original context in the Mishnah and its parallels. We shall examine how the dictum was understood in both Jewish and non-Jewish literature in order to trace the new meanings that were attached to it over the course of history in both traditions. METHODOLOGY: The article examines the Mishnaic source and its parallels in light of early manuscripts. Furthermore, it investigates the historical background and biographies of the many commentators on this dictum in order to account for the diverse explanations offered by those authors. RESULTS: An examination of the manuscripts demonstrates that this passage did not appear in the original Mishnah, but was added during a later period. The parallel texts imply that condemnation is limited to those physicians who treat their patients using heretical and idolatrous methods. Jewish commentators utilized this exceptional dictum as a means to teach important moral lessons, each author in accordance with his own viewpoint and the needs of the hour. Non-Jewish authors also exploited the passage, supporting their anti-Semitic ideology with Talmudic sources. CONCLUSION: No conclusions regarding the Jewish attitude toward medicine and doctors may be derived from the dictum: "The best of the physicians to Gehinom." The statement probably relates solely to doctors of that period who employed heretical and idolatrous practices in the treatment of their patients. The Talmud expresses its respect and appreciation for medicine and doctors. Jewish commentators across the generations continued to derive important moral lessons from this exceptional statement in order to enhance the professional and moral responsibility of Jewish physicians. PMID- 15523815 TI - [The ubiquitin system for intracellular protein degradation--involvement in human pathologies and therapeutic implications]. AB - The ubiquitin-proteasome pathway has a central role in selective degradation of intracellular proteins. Among the key proteins that are degraded by the system are those involved in the control of inflammation, cell cycle regulation, and gene expression. With so many important cellular pathways affected, derangements in the ubiquitin system have been shown to result in a variety of human diseases. Consequently, proteasome inhibition has a potential as a form of treatment for many human diseases such as cancer and inflammatory conditions. Two proteasome inhibitors, PS-341 and PS-519 are currently under clinical evaluation. PS-341 is currently being evaluated in phase III clinical trial for multiple myeloma, and PS-519 is now on a phase II trial for acute ischemic stroke. In addition, inhibition of the proteasome has been shown to be effective in several animal models for a variety of human diseases such as different malignancies, asthma, rheumatoid arthritis, and arterial restenosis. Future studies will be required to establish whether the promising animal studies could be successfully implicated in human disease states. PMID- 15523816 TI - [Vaccination policy in the IDF--past, present and future]. AB - The Army Health Branch of the Medical Corps in the Israel Defense Forces (IDF) maintains responsibility for the IDF's troop vaccination policy. This policy has undergone many changes and adaptations over time, due to shifting trends in civilian and military morbidity; technological advances and the introduction of new vaccines; changes to the cost-effectiveness ratio of specific vaccines; and other causes. The evolution of the IDF vaccination policy has not been reviewed to date, and can well serve as an example of the various considerations influencing public health policy in general, and specifically vaccination policy. Furthermore, questions are often raised by health care professionals as to discharged soldiers' military vaccination history, especially in the context of travel clinics. The authors present an historical review and current update of the IDF vaccination policy. PMID- 15523817 TI - [Cellular phones and public health]. AB - BACKGROUND: The increased use of mobile cellular phone by the public is associated with a wave of contradictory reports about the possible health effects, due to the exposure of the users to electromagnetic non-ionizing radiation. AIMS: This article reviews the state of the art of the present knowledge concerning the biological and medical effects of exposure to cellular phones, with an emphasis on its possible carcinogenic effect. RESULTS: Health conditions, which have been ascribed to the use of mobile phones mainly include some types of cancer and changes of brain activity. However, the balance of evidence from available studies has not yet supported these claims. Following the recommendation of special international expert committees, the IARC (International Association for Research on Cancer) is conducting a multi-center study to determine the possible effect of cellular phone use on brain and salivary gland tumors. Israel is one of the participants of this study. The only established health effect associated with the use of such technology is an increased risk for road accidents, unrelated to the amount of radiation emitted by phone. CONCLUSIONS: The challenge posed by this new technology to health authorities all over the world has lead to the definition of a new principle, the so-called "prudent avoidance", used as guidelines for the definition of an adequate public health policy. The public policy in Israel has used the prudent avoidance principles, while awaiting the results of the multi-national epidemiological studies. PMID- 15523818 TI - Hand movements deviate toward distracters in the absence of response competition. AB - In a series of experiments, participants reached to targets in the presence of visual distracters that were either adjacent to the target or located along the reach path. The reaching movements were affected by the presence of the distracters, with the movement paths deviating toward the distracters. Those deviations were observed under two different conditions: (a) one in which the distracter could potentially have been a movement target; and (b) another in which the distracter never was a possible target. Because the movement was affected by the distracter in both situations, the results suggested that response competition is not necessary for distracter-induced reach-path deviations. Instead, the authors propose that attention to a distracter is sufficient to affect the to-target movement. The movement deviations may reveal an effective mechanism for coping with stimulus-rich environments. PMID- 15523819 TI - Selective reaching: distracter effects on movement kinematics as a function of target-distracter separation. AB - The authors investigated distracter effects on the kinematics of reaching movements to determine when during reaching responses (reaction time, time to peak velocity, time after peak velocity, or peak velocity) distracter interference occurred and how target-distracter separation affected the locus of interference. Participants moved a pen on a digitizing tablet toward a target appearing with or without a distracter. With a small target-distracter separation, distracter interference occurred during time after peak velocity (similar amounts of interference from near and far distracters). With a large target-distracter separation, distracter interference occurred during time to peak velocity (more interference from near compared to far distracters). The results demonstrated that target-distracter separation is an important determinant of the locus of distracter interference. PMID- 15523820 TI - Space- and object-based attention depend on motor intention. AB - The authors investigated the impact of different motor demands on space- and object-based attention allocation. Responses to targets were either lifting a finger, or pointing to the target, or grasping a clay object placed on the target location. Reaction times and movement times were recorded to assess covert and overt attention, respectively. Both reaction times and movement times showed more space-based attention for pointing than for finger lifting and more object-based attention for grasping than for pointing. That result supports the view that visual selectivity is tuned to specific motor intentions (H. Bekkering & F. W. Neggers, 2002) and illustrates the tight coupling of perception and action. PMID- 15523821 TI - Visuomotor cuing through tool use in unilateral visual neglect. AB - In the present study, the authors examined the effect of tool use in a patient, MP, with neglect of peripersonal space. They found that target detection improved when the patient searched with his arm outstretched, when both visual and motor cues were present. Motor cues (arm outstretched but hidden from view) and visual cues alone (shining a torch on the objects) were less effective. In a final experiment, the authors reported that MP established a better memory for the objects that were searched for when a combined visual and motor cue was present. The authors argue that search was improved by combined visuomotor cuing, which was effective when the action could affect the objects present. Visuomotor cuing also led to stronger memories for searched locations, which reduced any tendency to reexamine positions that had been searched previously. The data are discussed in terms of the interaction between perception and action. PMID- 15523822 TI - Backward-compatibility effects with irrelevant stimulus-response overlap: the case of the SNARC effect. AB - In 3 dual-task experiments, the relationship between primary-task response (R1) and secondary-task response (R2) was varied. In general, R1-left responses were faster when followed by the word one, and right responses were faster when followed by the word two. This backward-compatibility (BWC) effect indicated (a) that activation of R2 was not delayed until R1 selection was completed, and (b) that activation of the vocal responses was accompanied by the automatic activation of magnitude codes, known to be associated with spatial left-right codes (spatial-numerical association of response codes [the SNARC effect]). These findings supported the hypotheses (a) that BWC effects persist even with irrelevant R1-R2 overlap, (b) that the SNARC effect is based on associations between magnitude and spatial representations underlying response processing, and (c) that the production and perception of magnitudes relies on common codes. PMID- 15523823 TI - Differences in attentional set between athletes and nonathletes. AB - The purpose of this study was to determine if the ability to set attention is different between athletes and nonathletes. University volleyball players were compared with nonathletes on a spatial cuing task similar to that of C. L. Folk, R. W. Remington, and J. C. Johnston (1992). As expected, both groups showed evidence of entering into a specific attentional set because attentional cuing effects were found only when cues and targets shared a task-relevant feature (onset or color). In addition, the cuing effects when the cues matched the targets were greater for the athletes than for the nonathletes. The results are discussed in terms of the orienting of attention and athletic performance. PMID- 15523824 TI - Illusory gravitational forces affect aimed limb movements. AB - The authors report on 2 experiments in which participants produced rapid wrist rotation movements in a clockwise direction to move a cursor from a home position into a target region. The display was constructed such that the target could be depicted above, below, or beside the home position, so that the clockwise wrist rotation could produce upward, downward, or rightward movements of the cursor. By parsing the movements into component submovements, the authors found that upward movements were consistently less forceful, as evidenced by shorter primary submovements, compared with downward and horizontal movements. Those results suggested a new virtual gravity illusion: Participants apparently overcompensated for the absence of the anticipated effects of gravity by adjusting the initial force they used to propel the cursor toward the target. PMID- 15523825 TI - What are you looking at? Acuity for triadic eye gaze. AB - The authors measured observers' ability to determine direction of gaze toward an object in space. In Experiment 1, they determined the difference threshold for determining whether a live "looker" was looking to the left or right of a target point. Acuity for eye direction was quite high (approximately 30 s arc). Viewing the movement of the looker's eyes did not improve acuity. When one of the looker's eyes was occluded, the observers' acuity was disrupted and their point of subjective equality was shifted away from the exposed eye. Experiment 2 was a replication of Experiment 1, but digitized gaze displays were used. The results of Experiment 3 showed that the acuity for direction of gaze depended on the position of the looker's target. Overall, the results indicated that humans are highly sensitive to gaze direction and that information from both eyes is used to determine direction of regard. PMID- 15523826 TI - Effect of furastanol glycosides of Dioscorea on lipid peroxidation in tomatoes infected with gall nematode. PMID- 15523827 TI - Fungicidal activity of phosphinic analogues of amino acids involved in methionine metabolism. PMID- 15523828 TI - Involvement of some carrier proteins in thermoregulatory enhancement of respiration of mitochondria of the liver and skeletal muscles of ground squirrels (Citellus undulatus) awakening from hibernation. PMID- 15523829 TI - RepA protein of the bacteriophage N15 exhibits activity of DNA helicase. PMID- 15523830 TI - Degree of homogeneity of cholinesterase activity of nervous tissue of squids as a taxonomic trait. PMID- 15523831 TI - Role of C- and N-terminal mutations of the movement protein of tobacco mosaic virus in activation of complexes between the transport protein and viral RNA that are not translated in vitro. PMID- 15523832 TI - Bacterial degradation of technogenic radioactive particles. PMID- 15523833 TI - Epibrassinolide treatment changes the polypeptide composition of buckwheat fruits. PMID- 15523834 TI - Protein sequences as a "literary" text. PMID- 15523835 TI - Cloning, expression, and purification of the nucleocapsid protein of SARS coronavirus. PMID- 15523836 TI - Molecular modeling of N-terminal domains of NMDA-receptor. Study of ligand binding to N-terminal domains. PMID- 15523837 TI - The unknown nuclear protein that decreases HNF3 activity ensures species specificity of action of hepatocarcinogenic amino azo dyes. PMID- 15523838 TI - Expression of ceruloplasmin pseudogene in cultured human cells. PMID- 15523839 TI - Cytokinin control of rRNA gene transcription in senescing barley leaves. PMID- 15523840 TI - [Evolution of mammalian sex chromosomes: cooperation of genetic and epigenetic factors]. AB - The X and Y chromosomes of mammals, which significantly differ in structure and genetic composition, are thought to originate from a pair of autosomes. During evolution of sex chromosomes in placental mammals, the degradation of the Y chromosome and inactivation spreading along the X chromosome occurred gradually and in concert. Thus, at the molecular level, the genetic and epigenetic factors interacted toward greater differentiation of the X/Y pair. In this review, in context of a comparison permitting to trace this evolutionary pathway, we consider the structural features of mammalian sex chromosomes focusing on the X chromosomal genes and the unique epigenetic mechanism of their regulation. Possible causes and consequences of the genes skipping inactivation and aspects of molecular mechanism of X-chromosome inactivation are discussed. A number of hypotheses are considered on evolutionary relationships of X-chromosome inactivation and other molecular processes in mammals. PMID- 15523841 TI - [Instability of repetitive units of foreign centromeric satellite DNA in transgenic mice and transfected cells]. AB - Cytologically detectable instability of centromeric satellite DNA may cause hereditary disorders in human. To study the mechanisms of such instability, two transgenic mouse lines and 11 clones of transfected F9 mouse embryonic teratocarcinoma cells were obtained with the 3.8-kb repetitive unit (Sat) of Bos taurus satellite DNA IV. Intergeneration and somatic instability of exogenous satellite DNA (satDNA) was observed in transgenic mice and transfected cells as a change in nucleotide sequence of an internal Sat region approximately 1000 bp in size. Since Sat was in the hemizygous state in both cases by the experimental protocol, the instability was attributed to intra-allelic processes. Intergeneration instability probably took place in the premeiotic period of gametogenesis or in early embryo development and led to prenatal death of transgenic embryos after at least one generation. No direct or inverse correlation was observed between methylation and instability of Sat. The results testify that submicroscopic changes in highly repetitive noncoding DNA sequences may already affect the genome function in higher eukaryotes. PMID- 15523842 TI - [Unequal crossing over as the pathway of adaptive homologous recombination between the direct DNA repeats in tandem duplications of Escherichia coli]. AB - Homologous recombination between direct DNA repeats within the extended tandem duplications in E. coli results from unequal sister-chromosome exchanges. This conclusion follows from the observations on the segregation of completely or partly homozygous diploid segregants by heterozygous duplications. The formation of diploid segregants with preserved heterozygosity for the unselected markers could also result from "symmetrical" intrachromosomal recombination. Analysis of the segregant genotypes, however, confirmed their formation via unequal crossing over. The data obtained indicated that in tandem duplications segregation of diploid recombinants of different types was preceded by the formation of triplications as the products of unequal sister-chromosome exchanges. In heterozygous duplications, unequal crossing over is manifested as a highly frequent adaptive exchange, providing the survival of the most part of the duplication-carrying cells on selective medium. It is suggested that adaptive mutagenesis can be the consequence of unequal sister crossing over. PMID- 15523843 TI - [The effect of isogenization on the phenotypic manifestation of quantitative traits in Drosophila melanogaster]. AB - A comparative analysis of the phenotypic values of the proximal and distal fragments of the radial wing vein was carried out in heterogeneous lines of Drosophila melanogaster and in isogenic lines derived from them with the help of a balancer line. The mean values of the traits in the isogenic lines were shown to significantly differ from the corresponding values in the "parental" heterogeneous lines. Apparently, the change in the trait values was caused by a double recombination exchange between the inverted and the "normal" chromosomes, which suggests partial crossing over suppression in the balancer lines. PMID- 15523844 TI - [Age dependence of the level of copia retrotransposon expression in the testes of Drosophila melanogaster]. AB - Expression of the lacZ reporter gene controlled by various deletion derivatives of the regulatory region of the copia retrotransposon was studied in the testes of adult transgenic males of the Drosophila melanogaster y1W67c23(2) strain at the age of 3, 6-9, 12-15, 18-21, and 24-27 days. When the construct contained the full-length regulatory region, which included the 5'-long terminal repeat (LTR) and the 5'-untranslated region (UTR), expression was the lowest in males aged 12 15 days and the highest in males aged 3 or 24-27 days. A similar V-shaped age dependence was previously observed for the copia transposition rate and RNA content in a strain with a high rate of copia transposition. Thus, the V-shaped age dependence of expression, which is unusual for Drosophila, proved to be characteristic of copia regardless of its transposition rate. Deletion of the 5' or 3' end of the LTR, but not of the UTR, changed the age dependence of the level of reporter gene expression. In this case, expression was the highest in 3-day old males and gradually decreased with age, as typical for many Drosophila genes. It was assumed that the 5'- and 3'-terminal regions of the copia LTR contain regulatory elements responsible for the V-shaped age dependence of expression, while the expression level depends to a greater extent on the regulatory elements of UTR. PMID- 15523845 TI - The induction of chromosomal aberrations by tetra antibiotic in bone marrow cells of rats in vivo. AB - The aim of this study was to investigate the in vivo effects of Tetra (Tetralet) antibiotic on the chromosomal aberrations (CA) in bone marrow cells of rats (Rattus norvegicus var. albinos). Tetra antibiotic significantly increased the percentage of abnormal cells and the chromosomal aberrations per cells (CA/cell) in bone marrow cells of rats at concentrations of 100 and 200 mg/kg body weight for 12 and 24 hours treatment periods for each. In addition, the percentage of abnormal cells and the CA/cell increased dose-dependently for 12 hours treatment period; In contrast, mitotic index (MI) was decreased when compared with negative control and solvent controls for 12 hours treatment period. However, MI increased depend on Tetra antibiotic dose for 24 hour treatment period. PMID- 15523846 TI - [Interspecific genetic variability of the oyster mushroom Pleurotus ostreatus as revealed by allozyme gene analysis]. AB - Allozyme variation in natural populations of basidiomycetes fungus Pleurotus ostreatus (88 individuals) from three regions of central Russia was studied. The species was shown to have 92.86% of polymorphic allozyme loci and expected heterozygosity He = 0.49. The mean number of alleles per locus was 3.5. The genetic differences among populations were supported by F-statistics (FST = 0.750). The low level of inbreeding (FIS = 0.018) suggests that the P. ostreatus populations are panmictic, and the main reproduction mode involves basidiospores dispersing at long distances. Using cluster analysis, geographically isolated populations and intersterile groups were differentiated within the complex P. ostreatus species. PMID- 15523847 TI - [Chromosome maps of trilliaceae: II. A study of the genome composition in polyploid species of the genus Trillium by fluorescence nucleotide base-specific staining of heterochromatic chromosome regions]. AB - Chromosome banding with nucleotide base-specific fluorochromes chromomycin A3 (CMA) and Hoechst 33258 (H33258) was used to study the karyotypes and to construct cytological maps for diploid Trillium camschatcense (2n = 10), tetraploid T. tschonoskii (2n = 20), hexaploid T. rhombifolium (2n = 30), and a triploid T. camschatcense x T. tschonoskii hybrid (T. x hagae, 2n = 15). With H33258, species- and genome-specific patterns with numerous AT-rich heterochromatin bands were obtained for each of the four forms; CMA revealed a few small, mostly telomeric GC-rich bands. In T. tschonoskii, the two subgenomes were similar to each other and differed from the T. camschatcense genome; on this evidence, the species was considered to be a segmental allotetraploid. In T. x hagae, one T. camschatcense and both T. tschonoskii subgenomes were identified. The subgenomes of T. rhombifolium only partly corresponded to the T. camschatcense and T. tschonoskii genomes, in contrast to the morphologically identical Japanese species T. hagae. This was assumed to indicate that allohexaploids T. rhombifolium and T. hagae originated independently at different times; i.e., their origin is polyphyletic. Based on the chromosome maps, a new nomenclature was proposed for the Trillium genomes examined: K1K1 for T. camschatcense, T1T1T2T2 for T. tschonoskii, T1T1T2T2 for T. x hagae, and K1RK1RT1RT1RT2RT2R for T. rhombifolium. PMID- 15523848 TI - [Analysis of microsatellite loci of the chloroplast genome in the genus Capsicum (Pepper)]. AB - Six plastome microsatellites were examined in 43 accessions of the genus Capsicum. In total, 33 allelic variants were detected. A specific haplotype of chloroplast DNA was identified for each Capsicum species. Species-specific allelic variants were found for most wild Capsicum species. The highest intraspecific variation was observed for the C. baccatum plastome. Low cpDNA polymorphism was characteristic of C. annuum: the cpSSRs were either monomorphic or dimorphic. The vast majority of C. annuum accessions each had alleles of one type. Another allele type was rare and occurred only in wild accessions. The results testified again to genetic conservation of C. annuum and especially its cultivated forms. The phylogenetic relationships established for the Capsicum species on the basis of plastome analysis were similar to those inferred from the morphological traits, isozyme patterns, and molecular analysis of the nuclear genome. PMID- 15523849 TI - [Allozyme variation of the pygmy wood mouse Sylvaemus uralensis (Rodentia, Muridae) and estimation of the divergence of its chromosome forms]. AB - The genetic divergence between the eastern European, southern European, and Asian chromosome forms of the pygmy wood mouse Sylvaemus uralensis, whose karyotypes differ from one another in the amount of pericentromeric heterochromatin, has been reevaluated using allozyme analysis. In general, Asian S. uralensis living in eastern Kazakhstan, eastern Turkmenistan (the Kugitang Ridge), and Uzbekistan are more monomorphic than European populations of this species. However, the allozyme differences between all chromosome forms of the pygmy wood mouse is comparable with the interpopulation differences within each form and are an order of magnitude smaller than those between "good" species of the genus Sylvaemus. Thus, the chromosome forms of S. uralensis cannot be considered to be separate species. The concept of races as large population groups that have not diverged enough to regard them as species but differ from one another in some genetic characters is used to describe the differentiation of S. uralensis forms more adequately. The currently available evidence suggests the existence of two S. uralensis races, the Asian and the European ones, and two chromosome forms (eastern and western) of the European race. The possible historical factors that have determined the formation of the races of the pygmy wood mouse are considered. According to the most plausible hypothesis, the shift and fragmentation of the broad-leaved forest zone during the most recent glacial period (late Pleistocene) were the crucial factors of the formation of these races, because they resulted in a prolonged isolation of the European and Asian population groups of S. uralensis from each other. PMID- 15523850 TI - [Far Eastern mullet Mugil soiuy Basilewsky (Mugilidae, Mugiliformes): the genetic structure of populations and its change under acclimatization]. AB - The introduction of Far Eastern mullet (pilengas) in the Azov Sea in the 1970s 1980s has resulted in the formation of a self-reproducing commercial population. We have carried out a comparative population-genetic analysis of the mullet from the native (Primorye, the Sea of Japan basin) and the new (The Azov Sea basin) ranges. Genetic characteristics of three Primorye and three Azov local samples were studied using electrophoretic analysis of 15 enzymes encoded by 21 gene loci. In the Azov mullet, the initial heterozygosity characteristic of the donor population was preserved while the genotype and the allele compositions changed; the changes included a 1.9-fold reduction in the percentage of polymorphic loci and 1.5-fold reduction in the mean number of alleles per locus. The genetic differences between the Azov and the Primorye sample groups were highly significant. In the native range, no genetic differentiation among the mullet samples from different areas was found (Gst = 0.42%), whereas in the Azov Sea basin, the samples from spatially isolated populations (ecological groups) exhibited genetic differences (Gst = 1.38). The genetic divergence of the subpopulations and the excess of heterozygotes at some loci in the Azov mullet suggest selection processes that formed genetically divergent groups associated with the areas of different salinity in the new range. The salinity level is assumed to be the most probable factor of local differentiating selection during fast adaptation and naturalization of the introduced mullet. PMID- 15523851 TI - [Analysis of the IT15 gene in Huntington's disease families]. AB - Direct molecular genetic testing carried out in 59 Huntington's disease patients belonging to 46 families from Bashkortostan revealed the (CAG)n repeat expansion in exon 1 of the IT15 gene in 57 of them. By use of this analysis the disease status was not confirmed in two patients with atypical form of the disease and negative family history. The (CAG)n repeat expansion was identified in 27 out of 127 asymptomatic at-risk individuals. Analysis of the mutant (CAG)n allele inheritance demonstrated extremely high instability and high mutation rate predominantly leading to the appearance of the alleles with increasing number of (CAG)n repeats in subsequent generations. The instability was mostly observed in cases of paternal transmission. Almost complete linkage disequilibrium between the (CCG)7 mutant alleles and the del2642 deletion was demonstrated. Three major haplotypes revealed, (CCG)7/del-, (CCG)7/del+, and (CCG)10/del-, implied the existence of at least three sources of the origin of Huntington's disease in Bashkortostan. The identified haplotype frequency distribution patterns displayed similarities with those in European populations. The contribution of a number of genetic factors to the age of onset of Huntington's disease was analyzed. PMID- 15523852 TI - [Genetic demographic analysis of western Ukrainian populations: the marriage structure of populations from the Khmel'nitskii oblast with respect to ethnicity and birthplace]. AB - Ukrainians account for 85 and 91% of the populations of the city of Khmel'nitskii and the town of Starokonstantinov (Khmel'nitskii oblast, western Ukraine), respectively, and for 97% of the rural population of the Khmel'nitskii oblast. The proportions of Russians in the urban and rural populations of the Khmel'nitskii oblast are 7-10 and 1%, respectively. Between 1960 and 1995-1998, the proportions of Ukrainians in all populations studied increased and the proportion of interethnic marriages steadily decreased. The marriage association coefficient (K) with respect to ethnicity varied from 0.35 to 0.76 in different years. The highest assortative marriage indices (A') with respect to ethnicity were 75-98 and 71-84% in Ukrainians and Jews, respectively. The migration coefficient was 0.58-0.77. Western Ukrainian populations differ from eastern Ukrainian ones in a steadily decreasing outbreeding component. PMID- 15523853 TI - [Epidemiology of congenital malformations in Gorno-Altaisk, Altai Republic, Russia]. AB - A retrospective epidemiological study has been performed using the data from healthcare institutions of the city of Gorno-Altaisk, Altai Republic, Russia for the period from 1983 to 2001. Congenital malformations (CMFs) have been studied in newborns, infants that died at ages under one year, and fetuses after 22 weeks of gestation. The most frequent malformations are those of the musculoskeletal and cardiovascular systems and multiple malformations, which account for 37.68, 18.22, and 8.9% of all congenital malformations, respectively. Their frequencies are 7.38, 3.57, and 1.74 per thousand, respectively. The frequency of congenital malformations subject to registration by the national system of CMF monitoring of the Russian Federation (21 malformation forms) is 6.08 per 1000 births and varies from 8.59 to 21.24. The frequency of the Down syndrome is 0.93 per 1000 births; it did not vary significantly during the period studied. The frequency of limb reduction deformities in the urban population of Altai Republic (0.32 per 1000 births) is higher than in other Siberian regions, including the cities of Kyzyl (Tyva Republic) and Tomsk and the Nyurba and Ust-Aldan uluses of Sakha Republic (Yakutia). PMID- 15523854 TI - [Normal and adverse genetic processes in subdivided populations of the island type: computer simulation]. AB - A computer model simulating genetic dynamics of a subdivided population at the level of a one-locus diallele system is proposed. In contrast to the earlier publications, this model is based on Wright's island model. The computer experiments were focused on genetic changes occurring during relatively short time intervals comparable to the duration of economic activities of humans (tens of generations). The process of attaining stationary state and reorganization of the genetic structure of the system under anthropogenic influence were simulated. The results are interpreted graphically as a spatial-temporal distribution of subpopulations by classes of gene frequencies as well as in quantitative terms of Nei's G-statistics. PMID- 15523855 TI - Uterine fibroid: a review. AB - BACKGROUND: Uterine fibroid is a benign tumour of uterine smooth muscle. The purpose of this review is to bring to light the current spectra of presentation and management status of this benign and very important cause of menstrual and fertility disturbance in African women and the Black race in general. This is especially so with the trivialization of its aetological factors and treatment by claims from alternative medical practitioners. This review, therefore sought to document what is currently known about the condition and what could possibly be done to achieve better results in its management. METHODS: Literature on the subject above was reviewed using manual library search, electronic books such as CD-ROMS and journals articles published by various local and international authors on the subject; it also included internet search on relevant aspects of the topic. RESULTS: Fibroid is the commonest benign tumour of the female genital tract, it contributes about 70 to 80% of new growths in the female genital tracts, it is a cause of significant morbidity in women of reproductive age group and when complicated could be a significant cause of mortality. Spectrum of presentation mainly involves disturbance of menstruation, reduction in fertility, pressure and obstructive symptoms and rarely malignant presentations. When recognized early effective conservative and definitive therapies are available to offer relief for women and to make their menstrual and reproductive life more meaningful. CONCLUSION: From the literature reviewed, it was obvious that the subject of uterine fibroid is still not fully exhausted and there were a lot of research questions waiting to be answered on the aetiopathogenesis of the disease. There is also the need to fashion out better treatment alternatives that will reduce morbidity in the process of managing the patient as well as those that will further enhance reproductive potentials after treatment. PMID- 15523856 TI - Glucocorticoids and the insulin resistance syndrome. AB - BACKGROUND: Insulin regulates the uptake, utilization and storage of cellular nutrients. An attenuated responsiveness of tissues to insulin (insulin resistance), results in the metabolic or insulin resistance syndrome, polycystic ovary disease, lipodystrophic states and leprechaunism. Type 2 diabetes mellitus, a consequence of insulin resistance and a major component of the insulin resistance syndrome, currently affects 177 million people worldwide and is predicted to rise to over 300 million by 2025 (the majority of cases in Africa). METHODS: We reviewed currently available literature on the role of glucocorticoids in the pathogenesis and clinical management of the insulin resistance syndrome. RESULTS: The epidemiology of insulin resistance, the components of insulin resistance syndrome, factors responsible for insulin resistance, the role of glucocorticoids, and the treatment of gluocorticoid induced insulin resistance have been discussed. CONCLUSION: Chronically elevated serum glucocorticoid levels contribute to diabetes and removal of glucocorticoid excess improves insulin sensitivity. Because insulin regulates 11beta hydroxysteroid dehydrogenase enzyme type-1 (11betaHSD-1) activity, it has been suggested that dysregulation of (11betaHSD-1) may underlie the pathogenesis of glucocorticoid-dependent insulin-resistance. Indeed, 11beta-HSD1 has been proposed as a new target for type 2 diabetes drugs. PMID- 15523857 TI - The prevalence of hepatitis B surface antigen (HBsAg) among prospective blood donors and patients in Port Harcourt, Nigeria. AB - BACKGROUND: HBV infection with its associated sequelae is a disease of major public health importance worldwide. This study analyzed the results of HBsAg screening at the University of Port Harcourt Teaching Hospital (UPTH), Port Harcourt, over a five-year period (1996-2000), with a view to establishing the prevalence rate in this area of the Niger Delta of Nigeria. METHODS: RESULTS from the Hepatitis Laboratory of the Hospital for the five-year period (1996-2000) were reviewed. The subjects consisted of two (2) main groups, i.e. blood donors and hospital patients. Screening was done using latex agglutination kits manufactured by Biotec Laboratories, USA. RESULTS: A total of 7226 samples were screened for HBsAg over the five-year period. The overall sero-positivity rate was found to be 4.98%. The carrier rate among blood donors was 1.57%, while among hospital patients, the prevalence rate was 28.37%. CONCLUSION: Hepatitis B surface antigenaemia is common in Port Harcourt and its environs. Therefore adequate screening of patients and blood donors and the institution of adequate public health measures is advocated in order to reduce the transmission of this virus. PMID- 15523858 TI - Dental care needs and demands in patients attending the dental hospital of the Obafemi Awolowo University Teaching Hospital's Complex Ile-Ife, Nigeria. AB - BACKGROUND: To compare the pattern of need and demand for dental care in settings where services are paid for through government sponsored insurance programs and out of pocket by individuals. METHODS: Study consisted of first visit patients attending the Dental Hospital. The assessment of normative treatment needs was done. The tooth based treatment needs were assessed by the WHO basic methods and the periodontal needs were assessed by the periodontal need systems PTNS. RESULTS: More males had sought treatment than females. In both sexes, most of the patients demanding treatment belonged to the 17-34 year age group. There was no statistically significant difference between age groups (P = 0.65). 55.4, 8.1 and 1.4 percents sought care for toothache, caries without concomitant pain and dental check-up respectively. Oral surgical care was considered necessary for the main complaint in 50.1% of cases. However, the age group 17-34 year old needed more restorative care than periodontal and the > 65 age group requested more extractions. CONCLUSION: The expressed need or demand for dental care falls short of the normative need. It should therefore be the aspiration of appropriate government ministry and health care providers to attempt converting normative needs into demand for care. PMID- 15523859 TI - Typhoid enteric perforation in north western Nigeria. AB - BACKGROUND: Typhoid perforation is the most important surgical complication of typhoid enteritis and is associated with high morbidity and mortality. AIMS AND OBJECTIVES: To determine the pattern and outcome of management of typhoid perforation in Aminu Kano University Teaching Hospital, Kano. METHOD: A retrospective Analysis of patients treated for typhoid perforation over a 6-year period. RESULTS: There were 47 patients: 35 males and 12 females, ratio 2.9 to 1. The patients were aged 4 years to 58 years (mean 18.9 years). Typhoid perforation occurred all the year round with a peak prevalence in September; Six (12.8%) patients perforated in the first week, 29 (61.7%) second week, and 12 (25.5%) third week, of illness. Single perforation was found in 91.5% of cases, and two to three perforations in 8.5%. Surgical treatment was by simple closure in 72.3%, wedge resection in 8.5%, ileal resection in 17.1% and right hemi-colectomy in 2.1%. Of the 41 survivors (87.2%), wound infection was the most common postoperative complication in 44.7% of cases. The mortality rate was 12.8% mostly due to overwhelming sepsis. CONCLUSION: Typhoid perforation requiring surgical intervention is still endemic in our subregion, and emphasis should be on preventive measures such as safe drinking water and appropriate sewage disposal, and typhoid vaccination. PMID- 15523861 TI - Chronic osteomyelitis of long bones: reasons for delay in presentation. AB - BACKGROUND: Chronic osteomyelitis is a common orthopaedic infection and a major medical problem in developing countries. Time is a critical factor in the evolution of the chronic disease as most of our cases are a progression from acute haematogenous osteomyelitis. Our patients present very late. OBJECTIVE: To identify the reasons for this delayed presentation and suggest possible solutions. METHOD: Over three years (2000-2003) in a multi-centre study (2000 2003), we prospectively evaluated patients with both clinical and radiological diagnosis of chronic osteomyelitis. Besides personal data, mode of onset, duration of illness, father's educational status and occupation and reasons for delayed presentation were documented using a questionnaire. Previous surgery was an exclusion criterion. RESULTS: One hundred and thirty-six patients comprising 81 males and 55 females with a mean age of 17.7(2-55) years were recruited. The mean duration between onset of disease and presentation for treatment was 13.3(2 120) months. Sixty-one (44.9%) presented to a health facility were misdiagnosed and inappropriately treated; ignorance was the reason in 37(27.2%) and lack of finances in 32(23.2%). There was no statistically significant association between the father's educational status and occupation and the attitude to hospital attendance. CONCLUSION: Misdiagnosis, ignorance and poverty are the reasons for delayed presentation in patients with chronic osteomyelitis in this environment. A high index of suspicion, accurate diagnosis and appropriate therapy offer the only hope in containing the acute phase. Continuing medical education, improvement in social and medical amenities and the national health insurance scheme are possible ways of overcoming undue delay in presentation for specialist consultation. PMID- 15523860 TI - Anorectal disorders requiring surgical treatment in the University of Port Harcourt Teaching Hospital, Port Harcourt. AB - BACKGROUND: The prevalence of anorectal disorders requiring surgical treatment has not been previously documented in the University of Port Harcourt Teaching Hospital (UPTH). AIM: To find out which of the anorectal disorders were most commonly managed surgically in the UPTH from January 1 1998 to December 31 2002. METHODS: The case notes and theatre records of all anorectal conditions treated surgically were studied retrospectively. They were analysed with respect to age, sex, type of lesion, duration of pathology before surgery, surgical treatment performed, hospital stay, complications and outcome. RESULTS: Seventy patients were so treated. Males outnumbered females in a ratio of 1.2:1. The commonest age groups treated were in the first, third and fourth decades of life. The commonest conditions treated were haemorrhoid, fissure-in-ano, imperforate anus and fistula in-ano. Apart from imperforate anus cases, the mean duration of symptoms before surgery was 18 1/2 months. Open haemorrhoidectomy, lateral internal sphincterotomy and fistulotomy/fistulectomy were themain procedures performed. The procedures for imperforate anus varied from simple serial anal dilatation to perineal pull through operations. Main complication encountered was post hemorrhoidectomy anal stenosis. The mean hospital stay was 16.9 days. Mortality rate was 1.4%. CONCLUSION: Haemorrhoid, fissure-in-ano, imperforate anus and fistula-in-ano were the commonest conditions treated surgically in UPTH over the 5 year period. PMID- 15523862 TI - The bacteriology of open fractures in Ile-Ife, Nigeria. AB - BACKGROUND: The objective of this study is to determine the pattern of bacterial isolates and antibiotic sensitivity profile in open fractures. Fifty-nine patients with open fractures of the lower limb long bones were studied. The majority of the cases were Gustilo and Anderson types II 21(35.6%), and IIIA 16(27.1%). METHOD: The patients were assessed by history taking, physical examination, and plain radiographs. Wound swabs were taken for bacteriological studies. Other necessary investigations were also done. During wound debridement open fractures were classified into Grades I, II and III using the Gustilo Anderson method of classification. RESULTS: The infection rate was 45.8%. Gram positive cocci and Gram-negative rods were isolated. On the whole Staphylococcus aureus 13(25%), and Coagulase-negative staphylococci (CONS) 14(26.9%), were the commonest organisms isolated. The commonest Gram-negative rods that were isolated from the wounds were Proteus mirabilis 9(17.3%), and Pseudomonas aeruginosa 8(15.4%). Multiple organisms were commonly isolated from the wounds. While all the organisms isolated showed very good sensitivity to gentamicin, cloxacillin and ofloxacin, most of the organisms were resistant to penicillin, ampicillin and tetracycline. The sensitivity ranged from 62.5% to 100%. Coagulase-negative Staphylococci (CONS) showed sensitivity to the widest range of antibiotics which included penicillin, ampicillin, chloramphenicol, gentamicin, cloxacillin and ofloxacin. The sensitivity ranged from 35.7% to 100%. There was statistically significant difference between the open tibial and femoral fracture cases with respect to the interval between injury and debridement time (p = 0.008); the rate of wound infection (p = 0.021); and the occurrence of osteomyelitis (p = 0.023). The commonest complications observed were wound infection 27(45.8%), and delayed fracture union 26(44.1%), which were commoner in the open tibial fractures. CONCLUSION: This study shows that Staphylococcus aureus and coagulase-negative staphylococci (CONS) were the commonest organisms associated with open fracture of the lower limbs in our centre and that delay in the initial wound debridement was a major predisposing factor to wound infection. PMID- 15523863 TI - HIV positive patients and the surgeon. AB - BACKGROUND: Guidelines for the surgical treatment of HIV infected patients are lacking in Nigeria. The Social stigma of the infection and fears of contracting the disease discourage healthcare providers from treating the patients. OBJECTIVE: To present cases treated by surgeons and thereby stimulate interest on the need to offer these patients needed treatment. METHODS: A written questionnaire was sent to surgeons based in the University of Port Harcourt Teaching Hospital who have operated on patients known preoperativly to have HIV. Information sought included age, gender, indication for surgery, operation done, outcome of operation, indication for HIV test and information given to the patient after the test. RESULTS: a total of 26 HIV infected patients received surgical treatment in private and company clinics and there were 18 females and 8 males aged between 8 and 48 years. Most of these were in the 2nd to 4th decades. The operations were elective, emergency or urgent. Pregnancy-related indications predominated. Three patients volunteered their status. One when told she was HIV positive declined a planned right nephrectomy for hypernephroma. Nineteen patiens were discharged from hospital well. Three patients and parents of two were informed by the surgeons that they were HIV positive preoperatively. Five patients died postoperatively. CONCLUSION: There is need to promote awareness on the obligation of suregeons to offer these patients surgical treatment. A protocol and adequate facilities for prophylaxis for health care workers exposed to infection from patients should be established in Nigeria. PMID- 15523864 TI - Chronic gastritis and Helicobacter pylori infection in University College Hospital Ibadan, Nigeria--a study of 85 fibre optic gastric biopsies. AB - BACKGROUND: Fibreoptic endoscopic gastric biopsy, has revealed that chronic gastritis is not only of a complex aetiological background, but also varied topographical distribution and other clinico-pathological associations. By far, the most important aetiologic association of chronic gastritis is chronic infection by the bacillus Helicobacter pylori. This study aims to highlight some of the histopathological associations between this bacterium and chronic gastritis. METHOD: We reviewed all endoscopic gastric biopsies with a histological diagnosis of chronic gastritis, with available case records and archival tissues over an 18 year period, to determine association between the various pathological features of chronic gastritis present and the presence of H. pylori infection. RESULTS: Eighty-five cases with histological diagnosis of chronic gastritis fulfilled the inclusion criteria, and Helicobacter pylori infection was found in 22.4% of them. Most patients (83.1%) with chronic gastritis had moderate to severe grades of inflammation, and most (84.2%) of the infection involved the gastric antrum. Pangastritis was uncommon (10.5%). A greater proportion of the antral biopsies (87.5%) showed moderate to severe grades of chronic gastritis as compared with 16.7% of corporal biopsies. There was an early age of acquisition of infection, starting at the second decade of life, with a peak in the 6th decade. The frequencies of atrophic gastritis and intestinal metaplasia seen complicating chronic gastritis were 16.7% and 9.4% respectively. CONCLUSION: The study shows that Helicobacter pylori infection is associated mainly with moderate to severe chronic gastritis in Nigerians. This study confirms the antral predilection of H. pylori infection, and the finding of an antral preponderance for chronic gastritis partly explains the relatively more frequent occurrence of duodenal ulcers in Nigerians as compared to gastric ulcers. Furthermore, the finding of a low prevalence of atrophic gastritis and intestinal metaplasia, as well as the rarity of pangastritis in our patients probably explains in part, the low incidence of gastric cancer in Nigeria. PMID- 15523865 TI - Use and abuse of analgesics in Nigeria: a community survey. AB - BACKGROUND: Chronic consumption of analgesics has been shown to increase the risk of end-stage renal disease. In Nigeria, these drugs are readily available as there is no legislation regulating their consumption. We report the use and abuse of analgesics in a Nigerian population. METHOD: Using a pre-tested questionnaire, subjects in Zawan B ward in Jos were interviewed regarding analgesic use. Cumulative lifetime dose of drugs consumed was calculated by obtaining the product of the average number of pills consumed in a week by duration of use in years. Analgesic abuse was defined as cumulative lifetime use of >5000 pills. RESULTS: Common analgesics regularly consumed were paracetamol in 58.1%, analgesic mixtures 28.9% and NSAIDS 13%. Common indications for use of these drugs were musculoskeletal pains, headache, fever and stress. Analgesic abuse was present in 22.6% of the respondents. CONCLUSION: Analgesic abuse is common in Jos, Nigeria. The sale and consumption of these drugs should be restricted by legislation. PMID- 15523866 TI - Effect of socio-economic status, age and sex on antibody titre profile to Salmonella typhi/paratyphi in Ile-Ife, Nigeria. AB - BACKGROUND: Typhoid fever remains a great socio-medical problem in many poor and underdeveloped countries, with an estimated 16 million cases and 600,000 deaths occurring each year. Although it has a world wide distribution, typhoid fever is endemic where sanitary controls are inadequate. The distribution of the infection in relation to age and sex has shown that typhoid fever is more prevalent in males than in females with out satisfactory explanation. Also, those in their second and third decades are more susceptible. The aim of the study is to evaluate the effect of socio-economic status, age and sex on antibody titre profile to Salmonella typhi and praratyphi in Ile-Ife, using both healthy individuals with immunological scar to Salmonella typhi/paratyphi and enteric fever patients. METHOD: Healthy volunteers (310) were recruited into the study and clinically diagnosed/culture proven cases (48) were used as control. Widal text was performed on each serum. Slide agglutination was first done, then positive samples were further subjected to tube agglutination for quantitative titration. RESULTS: The relationship between age and antibody titres in both healthy subjects and enteric fever patients is not statistically significant, (P values > 0.05). From the study also, sex and social status do not influence antibody titres to salmonella typhi/paratyphi (P value > 0.05). CONCLUSION: The study has shown that age, sex and social class do not statistically influence antibody titre profile to S. typhi/paratyphi in Ile-Ife, south western Nigeria. PMID- 15523867 TI - Tuberculosis in Rivers state: autopsy and surgical pathology study in the University of Port Harcourt Teaching Hospital (UPTH), Port Harcourt (1990-2002). AB - BACKGROUND: A 13 year (1st January 1990 - 31st December 2002) retrospective study of 138 surgical and autopsy specimens diagnosed as Tuberculosis (Tb) was studied in Port Harcourt, based on the age, sex, affected tissue and the immune status of the patients. METHOD: The tissues were initially fixed in 10% formal saline, processed and embedded in paraffin wax. The tissue were sectioned, mounted on glass slide and stained with hematoxylin and eosin (H & E). In some cases (12.3%) special stains were used to confirm the bacilli and to exclude parasitic or fungal agents. The immune status and the causes of death were extracted from the request forms, case notes and autopsy reports. RESULTS: Of the 138 specimens, 37 (26.8%) and 101(73.2%) were obtained from autopsy and surgery respectively. Male to female ratio was 2:1. The youngest was a 3 year old male and the eldest was a 83 year old female. Children below 11 years contributed the highest specimens at autopsy while the highest frequency of surgical specimens were removed from 41-50 years age group (23.8%). The commonest cause of death was pulmonary tuberculosis with respiratory failure. The commonest tissue affected was the lymph node presenting either singly or matted together. Primary tuberculosis accounted for 15.9% (22) of cases while secondary tuberculosis was 116 (84.1%) presenting in different forms. HIV screening was positive in 39 (28.3%) cases. CONCLUSION: Despite the preventive measures and treatment of tuberculosis, the disease is still on the increase in this environment. The upsurge may be associated with HIV infection and immunodeficiency. It is therefore necessary to screen all tuberculosis patients for possible HIV positivity. PMID- 15523868 TI - HIV sero-prevalence among newly diagnosed adult pulmonary tuberculosis patients in Sagamu. AB - BACKGROUND: [corrected] The association between Tuberculosis (TB) and Human immunodeficiency virus (HIV) has been described in several studies. TB was well represented in the first description of AIDS in Africa. In 1999, 25% of TB cases were attributable to the spread of HIV/AIDS. Frequencies of between 54-79% were recorded in medical in-patients in our hospital. The objective of the study is to assess the frequency of HIV in TB outpatients attending the tuberculosis and leprosy control centre in Sagamu. METHODS: A cross-sectional study of adult tuberculosis patients admitted into the DOTS programme of the TB and leprosy control centre, Olabisi Onabanjo University Teaching Hospital, Sagamu between January 2001 and December 2002 was undertaken. Free anti tuberculosis drugs were provided by the German leprosy Relief Association. Data was collected using interviewer-administered questionnaire. HIV screening, packed cell volume and sputum microscopy for acid fast bacilli were carried out on all patients. RESULTS: A total of 269 cases of pulmonary T.B were registered at the clinic of which 40 (14.9%) were HIV sero-positive. The peak age prevalence was in the 3d decade accounting for 42.5% of cases, followed by 32.5% in the 2rd decade. There was a statistically significant difference in the bacillary count and packed cell volume between HIV positive and negative TB patients. The mean weight at presentation, sputum positivity rate and sputum conversion rate at the end of two months of therapy was similar in both HIV positive and negative TB patients. CONCLUSION: This study has demonstrated a frequency of 14.9% of HIV sero prevalence amongst our TB population. Efforts should be intensified to ensure adherence to INH prophylaxis among HIV infected individuals in TB endemic areas as well as strengthening preventive measures. PMID- 15523869 TI - Survey of knowledge of HIV infection among senior secondary school 3 (SSS3) students in Port Harcourt. AB - BACKGROUND: The objective of this paper is to study the knowledge of the students on Human Immunodeficiency Virus and Acquired Immunodeficiency Deficiency Disease (HIV/AIDS) among third year senior secondary school (SSS3) students in Port Harcourt metropolis. METHOD: This was a cross-sectional survey involving 1800 SSS3 students from thirteen secondary schools in Port Harcourt metropolis, selected by a two-stage stratified sampling technique, using a self administered structured questionnaire. RESULTS: Thirty one percent, 14.4%, 9.1% and 8% of the students studied identified sexual intercourse, blood transfusion, mother to child (vertical) transmission and intravenous drug use, respectively, as modes of transmission of HIV infection. Only 7.1% identified all the listed four modes of transmission of HIV whilst 0.7% of the students identified all the listed preventive methods. The level of knowledge of the students on HIV did not significantly increase with age (p = 0.745). Students from Federal school demonstrated more knowledge than students from state and private schools. Similarly, students from all-male school had more knowledge than students from all-female and co-educational schools. Students from high social class demonstrated more knowledge than those from middle and low social class. CONCLUSION: It is concluded that the knowledge of the students on the transmission and prevention of HIV/AIDS is poor. There is need to intensify HIV/AIDS education programmes among adolescents. PMID- 15523870 TI - An assessment of aspirin use in a Nigerian diabetes outpatient clinic. AB - BACKGROUND: We have conducted this study to assess the use of aspirin among adult diabetic outpatients in our hospital. METHODS: The records of all patients attending the weekly Diabetes clinic of the Wesley Guild Hospital (WGH), Ilesa, Osun state, Nigeria over one month were reviewed and aspirin use evaluated in light of the American Diabetes Association position statement (2003) on aspirin therapy in diabetes. RESULTS: Eighty-two patients in all were studied. Fourty three (52.4%) were males, 39 (47.6%) were females. Their mean age was 59.1 +/- 10.7 yrs (range 31-81). All were type 2 and had been diabetic for a mean of 5.2 +/- 5.7 yrs (1-26yrs). Concurrent hypertension, another major risk factor for cardiovascular disease was found in 71.9% and 12.2% were obese. Aspirin use was contraindicated in 1.2%. All other patients had at least one indication for the use of aspirin based on the ADA criteria but only 39% were taking aspirin regularly. CONCLUSION: The results of this present study suggest that aspirin is still grossly under utilised in clinic patients with diabetes despite proven benefits. There is need to stimulate awareness amongst health care providers. PMID- 15523871 TI - Suicide attempts in HIV/AIDS patients: report of two cases presenting with penetrating abdominal injuries. AB - BACKGROUND: We present two patients to highlight an emerging trend of suicide attempts presenting to the surgeons with penetrating abdominal injuries found to be HIV/AIDS related. METHODS: The two patients were managed at the University of Maiduguri Teaching Hospital in the year 2003. Both patients presented as emergencies with penetrating abdominal injuries and were resuscitated followed by clinical evaluation and laparotomy. RESULTS: Both patients had visceral injuries that were repaired at laparotomy and were both confirmed HIV positive. CONCLUSION: There is an emerging trend of suicide attempts in HIV/AIDS patients and the need to determine the suicide pattern that will enable all stakeholders workout a formidable HIV/AIDS and suicide prevention programmes. PMID- 15523872 TI - Unplanned vaginal birth after two previous caesarean sections. AB - BACKGROUND: Caesarean section is carried out for the benefit of both mother and baby. Most obstetricians will permit vaginal birth after a previous caesarean. An unplanned vaginal birth after two previous caesareans section is reported. METHODS: A case report utilizing the case records and review of relevant literature. RESULT: A 32-year old housewife gravida3, para(2+0), both alive with two previous caesarean sections had a premature rupture of membranes and preterm labour. She had an unplanned successful vaginal birth. CONCLUSION: Vaginal delivery is possible after two previous caesarean sections if careful selection of patients is made. PMID- 15523873 TI - Problems of postgraduate medical training in Nigeria. AB - BACKGROUND: Postgraduate medical training in Nigeria has been in dire straits for about two decades now. That it has continued to survive, is a tribute to the average resident doctor who has become immunized, and who has grown a thick skin of armour, as well the spirit of altruism of the medical teachers--consultants (young and old), who despite odds, have kept their focus clear, above the murky waters of national distraction and daunting socio economic challenges. METHOD: A review of relevant literature on medical education in Nigeria was undertaken by manual library search. RESULTS: This paper x-rays the strong points that have still prevailed to hold the rudiments and ideals of postgraduate medical training in a viable position up to date. It discusses the weaknesses and threats- potential and real--to the training programme. CONCLUSION: This paper attempts to search, and actually hopes, for silver lining in the Nigerian sky as possible solution lifelines that may yet re-engineer the programme. PMID- 15523874 TI - Clinical photographic quiz. PMID- 15523875 TI - Ameloblastoma: the most common odontogenic tumour? PMID- 15523876 TI - [Some issues of forensic medical expertise within civil processes in the light of provisions of the new civil procedural code of the Russian Federation]. PMID- 15523877 TI - [A modern system of procedural forensic medicine in criminal procedures of the Russian Federation]. PMID- 15523878 TI - [The forensic medical service in Germany: its stucture and scientific society of legal medicine]. PMID- 15523879 TI - [Expert evaluation of the gunshot distance based on the volume of wound canal in flat bones]. AB - Different-distance experimental shots were made from a rifled gun at flat bones. Those shot distances were established, at which the wound canal had the smallest volume, and, at which entry and exit holes were virtually identical and matched the bullet diameter. A bigger shot distance was associated with an increasing volume of the wound canal caused by a bigger exit hole. A smaller shot distance led, also, to an increasing volume of the wound canal caused by bigger entry and exit holes. The volume of the wound canal in flat bones can be used as a reliable criterion in assessing the shot distance. PMID- 15523880 TI - [The morphological specificity of relaxation of cardiomyocytes and their value in forensic medical diagnostics]. AB - The morphological specificity of different conditions of cardiomyocytes was studied in myocardia of 50 cadavers of men who died of a variety of reasons. All conditions, except for the normal relaxation of cardipmyocytes, were found to result from damages in the contractive apparatus. Morphological variations of relaxation of cardiomyocytes are described. PMID- 15523881 TI - [The activity of ethanol- and aldehyde-oxidizing enzymes of the brain in ethanol intoxication]. AB - The distributed activity of alcohol dehydrogenase (ADG) and aldehyde dehydrogenase (AldDG) was histochemically studied in neurons of locus coeruleus, nuclei of raphe, substantia nigra and in cerebellar cortex in death or intoxication of ethanol (IE). The enzymatic activity was found to be changing depending on whether there was IE or not. Both higher ADG and lower AldDG were registered in the examined brain sections, which differ by mediatory structure of the neuronic centers. The detected regularities can be used within the forensic medical diagnosis of IE. PMID- 15523882 TI - [The forensic medical diagnosis of intoxication of alcohol surrogates by morphological findings]. AB - The pathomorphology of intoxication of alcohol surrogates is described. A high frequency rate of DIC-syndrome is shown for the condition. A majority of thrombi accumulates at places with the highest concentrations of toxic substances as well as at those places, through which poisoning substances are brought out of the body, i.e. in the liver (since toxic substances are introduced perorally) and kidneys. Thrombi were detected in all internals, including the brain, in lethal intoxication. The immediate toxic effect from methanol and from higher spirits can matter in the genesis of changes, including acute swelling and chromatolysis with subsequent cell death. Exclusively cerebral or coagulopathic-cerebral types of thanatogenesis were registered in studied case of intoxication. PMID- 15523883 TI - [The state-of-the art of forensic medical investigation of conclusive proof and outlooks]. PMID- 15523884 TI - [Determination of somatotype of man in cranio-facial personality identification]. AB - On the basis of their independent research and through the analysis of published data the authors suggested quantitative criteria for the diagnosis of a somatotype of man by the dimensional features of the face and skull. M. A. Negasheva method, based on the discriminative analysis of 7 measurement features, was used in the individual diagnosis of a somatotype by V. V. Bunaka scheme (somatotypes-pectoral, muscular, abdominal and indefinite). The authors suggest 2 diagnostic models based on the linear and discriminative analysis of 11 and 7 measurement features for the skull. The diagnostic accuracy in case of main male som-atotypes makes 87 and 64.4%, respectively, with the canonic correlations of 0.574 and 0.292. The designed methods can be used in forensic medicine for the cranio-facial and portrait expertise. PMID- 15523885 TI - [Practical use of the molecular-genetic technologies in solving the tasks of forensic medical identification of remains in emergencies with huge human death toll]. AB - Previously, we have developed different variants, for the expert use, of information technologies, i.e. of computer automated analytical systems, to ensure an effective analysis of huge bulwarks of molecular-genetic expert findings. The results obtained in the above research significantly advanced the possibilities of expert evaluation related with personality identification under the conditions of a big death toll; they also cleaned the ground for the introduction of such methodological approaches in the practice of investigations of emergency accidents involving huge death tolls. Described within the present case study is a practical experience of forensic medical identification of unrecognized remains in air crashes with huge human death tolls implemented through computer-assisted complex molecular-genetic tests on the basis of indirect DNA identification involving the establishment of blood relationship. PMID- 15523886 TI - [Etalon-free laser mass-spectrometry as a new method of the elementary analysis of bone tissue at identification]. AB - The results of an experimental study dealing with the elementary composition of bone tissue by using the method of laser mass spectrometry are described in the paper. The method ensures the quantification of concentration values of all elements from the Mendeleev periodic table that can be made within an extensive dynamic range, which makes it promising in the forensic medical identification. PMID- 15523887 TI - [A death case of eugenol intoxication]. PMID- 15523888 TI - Ets2 transcription factor inhibits mineralization and affects target gene expression during osteoblast maturation. AB - Our goal is to understand how Ets family transcription factors affect the genetic programs that control bone development. Modest overexpression of Ets2 in transgenic mice leads to Down's syndrome-like bone abnormalities. We observed that in the MC3T3-E1 in vitro model of osteoblast development, mature osteoblasts have very high levels of Ets2 relative to the immature preosteoblasts. We hypothesized that overexpression of Ets2 could have noticeable effects on gene expression, and found that exogenous Ets2 expression results in a complete lack of mineralized matrix in stable Ets2 transfected cells. Our cDNA microarray-based expression profiling of preosteoblasts vs. differentiated osteoblasts revealed several genes previously unrecognized as having roles in osteoblast maturation and up-regulated only in the mature osteoblasts. The promoters of these genes and known osteoblast marker genes were examined for Ets transcription factor binding sites (EBSs). Interactions of these sites with Ets2 protein were tested by EMSA. In vitro expressed Ets2 protein was able to form a protein:DNA complex with both known (Bsp, Opn, ON) and novel (Btg2, CysC, Lum) bone-related genes. In addition, Cbfa1 was found to interact with Ets2, forming a complex on the Opn promoter. PMID- 15523889 TI - Vascular endothelial growth factor as a survival factor in tumor-associated angiogenesis. AB - This review focuses on vascular endothelial growth factors (VEGF), a group of structurally-related proteins that serve as key growth factors for tumor associated angiogenesis. Pathways induced by VEGF proteins that regulate biological functions of key cell types involved in tumor angiogenesis, including vascular endothelial cells, pericytes and tumor cells, are discussed. Strategies that are currently being developed and tested based on the emerging definitions of the roles of the multiple cell types involved in tumor vessel development, their selective production of VEGF-related proteins and other pro-angiogenic growth factors, their expression of associated receptors as well as identification of signal transduction pathways involved in VEGF-induced tumor survival and tumor-associated angiogenesis will be reviewed. PMID- 15523890 TI - Effect of n-3 fatty acids on the antitumour effects of cytotoxic drugs. AB - BACKGROUND: n-3 fatty acids are increasingly being administered to cancer patients for the treatment of cachexia, and it is thus important to know of any potential interactions with ongoing cytotoxic drug therapy. MATERIALS AND METHODS: For this reason eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were administered to mice bearing the cachexia-inducing MAC16 colon adenocarcinoma, and the effect of epothilone, gemcitabine, 5-fluorouracil and cyclophosphamide on tumour growth and body weight determined. RESULTS: Epothilone alone had a minimal effect on tumour growth rate, but this was potentiated by DH4, while for 5-fluorouracil and cyclophosphamide tumour growth inhibition was enhanced by EPA. The antitumour effect of gemcitabine was not altered by either fatty acid. EPA arrested the development of cachexia, while DHA had no effect and the same was true for their effect on tumour growth rate. The anticachectic effect of EPA was only seen in combination with 5-fluorouracil. CONCLUSION: These results suggest that n-3 fatty acids do not interfere with the action of chemotherapy and may potentiate the effect of certain agents. PMID- 15523892 TI - Leiomyosarcoma of the scrotum arising from the dartos muscle: a rare clinicopathological entity. AB - Scrotal leiomyosarcoma is a rare tumour arising from the dartos layer: We describe a case of scrotal leiomyosarcoma in a 40-year-old man. The patient was treated by a wide surgical excision and no recurrence has been recognized 36 months later. A review of the literature is presented, summarizing the principal clinical and morphological characteristics of this rare tumor. PMID- 15523891 TI - Cutaneus pseudolymphoma: a case report. AB - The authors report on a case of pseudolymphoma cutis in a 48-year-old man. The clinical and histopathological characteristics of this benign skin disorder, especially regarding the differential diagnosis with cutaneous B or T cell lymphomas, are reviewed. Finally, the use of hydroxychloroquine sulfate is suggested for the therapy of pseudolymphoma cutis, especially when the causal factor is unknown. PMID- 15523894 TI - 1-amino-4-phenyl-1,2,3,6-tetrahydropyridine and 1-amino-4-phenylpyridinium salt, the 1-amino analogues of neurotoxins, MPTP and MPP+, induce apoptosis in PC12 cells: detection of apoptotic cells by Comet assay and flow cytometric analysis. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is known to induce parkinsonism in humans when it is oxidized to the 1-methyl-4-phenylpyridinium salt (MPP+). We previously reported the syntheses of 1-amino-4-phenyl-1,2,3,6 tetrahydropyridine (APTP) and 1-amino-4-phenyl-pyridinium salt (APP+), the 1 amino analogues of the dopaminergic neurotoxins, MPTP and MPP+, respectively, and demonstrated that both APTP and APP+ are cytotoxic to PC12 cells. In this study, we found that both APTP and APP+ induce apoptotic cell death in PC12 cells. Apoptosis was determined by the Comet assay and flow cytometric analysis. Prior to using the Comet assay for detection of apoptotic PC12 cells, Comet images of apoptotic and necrotic cells were first distinguished by using several standards. Comet images were classified into four groups (A to D) according to their shapes. Class D consisted of the apoptotic cells and was easily distinguished. We also demonstrated that apoptotic and necrotic PC12 cells can be easily differentiated and quantified using the convenient Comet assay. PMID- 15523893 TI - P53 overexpression in nasopharyngeal carcinoma. AB - Nasopharyngeal carcinoma (NPC) is a characteristic tumor displaying epidemiological, genetic and regional distribution properties and is unique by its natural behavior and therapy. Investigation of the molecular and biological changes, gene amplifications and activations that occur during carcinogenesis and progression can provide new insight into the pathology of the disease and may add biological factors that can be used as new prognostic markers. The p53 tumor suppressor gene is the most frequently mutated gene in human cancer. Although point mutations in the p53 gene are observed in nasopharyngeal cancer, the mutation rate is lower than in other tumors. Immunohistochemical studies have shown significant p53 overexpression in NPC material. In this study, p53 protein immunoreactivity was investigated in paraffin sections of primary nasopharyngeal tumors and metastatic cervical lymph nodes and association with clinical and histopathological characteristics was evaluated. Ninety-seven paraffin sections from 81 patients with NPC treated from 1990 to 1996 were examined by immunohistochemistry and were correlated with clinical features and treatment outcome. Among a total of 97 samples, positive staining for p53 protein was observed in 83 (85.5%) samples while no staining was found in 14 (14.5%) cases. Immunoreactivity was observed in 62 (81.5%) of the primary nasopharyngeal biopsy specimens. The correlation between p53 expression and histological type, stage, age and sex distributions was tested. After statistical analysis according to Chi square test and Yates' correction, no significant difference was demonstrated (p>0.05). There was no statistically significant correlation with p53 immunoreactivity and overall and disease-free survival. Although the association between NPC and p53 is not clear, our study confirms that p53 overexpression is present in a considerable subset of patients with NPC. PMID- 15523895 TI - Analysis of the temporal events in blood and tissues following fecal peritonitis in mice. AB - BACKGROUND: The kinetics of systemic responses triggered by bacterial peritonitis were analysed in the blood, bronchoalveolar lavage (BAL) and several organs. MATERIALS AND METHODS: The murine model of cecal ligation and puncture was employed. The parameters were analysed at different periods of time (4 to 72 h). RESULTS: Fecal peritonitis triggered a progressive, but not significant, decline of blood leukocytes between 4 and 6 h, becoming significant at 24 to 48 h (p<0.05). This profile was closely associated with the enhancement of leukocytes both in the abdomen (p<0.05) and in the BAL (p<0.05). A significant abdominal exudation was detected between 4 and 72 h (p<0.01), whereas maximal growth of aerobic bacteria in the blood and lungs was observed 24 and 72 h after. Maximal exudation in the studied tissues occurred at different time points (heart=24 h, spleen and kidney=48 h, liver and lung= 72 h). CONCLUSION: Using this model, evidence of sepsis can be easily measured in different body systems. PMID- 15523896 TI - Management protocol for scirrhous gastric cancer. AB - BACKGROUND: The prognosis for patients with scirrhous gastric cancer (SGC) is extremely poor. However, the management protocol for this type of cancer has not been well discussed. In this study, we retrospectively evaluated the management of SGC and we introduced a new treatment protocol for SGC. PATIENTS AND METHODS: Between 1991 and 2001, 58 patients with SGC were treated. Thirty-nine, who underwent gastrectomy, were divided into 3 sub-groups according to peritoneal metastasis (P) and peritoneal washing cytology (CY) status [P(-)/CY(-), P( )/CY(+), and P(+)/CY(+)]. The survival rates of these 3 sub-groups were compared with patients who did not have a gastrectomy (n=19), retrospectively. From 2002, we started a new treatment protocol for SGC. Laparoscopic diagnosis of P or CY and intraperitoneal chemotherapy (IPC) were performed before performing laparotomy on 10 patients with SGC. RESULTS: The 5-year survival rate of the 19 patients in P(-)/CY(-) was 11.6%. The survival rates of patients with P(-)/CY(+) or patients with P(+)/CY(+) were no different from patients who did not have gastrectomy (pleural effusion or ascites negative). In 10 patients who were treated with the new protocol, 7 with P(-)/CY(-) underwent gastrectomy after IPC and 3 with P(+)/CY(+) underwent repeated IPC. CONCLUSION: Gastrectomy may not have prognostic benefit for patients with SGC with CY(+). Thus, we recommend laparoscopic diagnosis of peritoneal metastasis or peritoneal cytology before performing laparotomy on these patients. PMID- 15523897 TI - Biphasic effect of relaxin, inhibitable by a collagenase inhibitor, on the strength of human fetal membranes. AB - BACKGROUND: Relaxin has been proposed as a hormone involved in the collagen remodeling of the utero-placental unit. MATERIALS AND METHODS: Human fetal membrane explants were incubated with H1 or H2 relaxin for 48 hours and stretched until rupture in a materials testing machine. Co-incubation with a synthetic collagenase inhibitor was performed in order to examine whether the effects of relaxin could be inhibited. The effects on hydroxyproline and histology were evaluated. RESULTS: Biomechanical testing showed that H2 relaxin induced a biphasic weakening of human fetal membranes, an effect that was abolished after co-incubation with a collagenase inhibitor. H1 relaxin produced no significant effects on the biomechanical properties. The effects of H2 relaxin on the biomechanical properties were, however, not followed by changes in the hydroxyproline concentration or the histology. CONCLUSION: H2 relaxin had an effect on human fetal membranes and this effect may be mediated through collagenases. PMID- 15523898 TI - Effect of low-level laser therapy on osteoarthropathy in rabbit. AB - The aim of this study was to determine whether low-level laser therapy (LLLT) aided the recovery of damaged articular cartilage in joints with artificially induced osteoarthropathy (OA). OA was induced by injecting hydrogen peroxide (H2O2) into the articular spaces of both knees in rabbits, twice a week for 4 weeks. The induction of OA and the effect of LLLT were evaluated by biochemical, radiological and histopathological analysis. Superoxide dismutase (SOD) activity increased about 40% in the OA group, as compared to the controls. Although SOD activity in the OA group was not significantly different from the 2-week groups, it was significantly different from the 4-week control and treatment groups. There was also a significant difference between the 4-week control and treatment groups. Simple radiographs and three-dimensional computed tomographs (3D CT) did not show detectable arthropathy in the OA group, nor any particular changes in the 2-week groups. In contrast, distinct erosions were seen in the distal articular cartilage of the femur, with irregularity of the articular surface, in the 4-week control group, while the erosions were reduced and arthropathy improved slightly in the 4-week treatment group. Grossly, erosions formed on the articular surface in the OA group. In comparison, severe erosions damaged the articular cartilage in the 4-week control group, but not in the 2-week control and treatment groups. Regeneration of articular cartilage was seen in gross observations in the 4-week treatment group. Histopathologically, there was slight irregularity of the articular surface and necrosis in the OA group, and serious cartilage damage, despite slight chondrocyte regeneration, in the 4-week control group. Conversely, the 4-week treatment group showed chondrocyte replacement, with sometimes close to normal articular cartilage on the articular surface. These results suggest that LLLT was effective in the treatment of chemically induced OA. PMID- 15523899 TI - Immunocytochemical detection of members of the caspase cascade of apoptosis in high-grade astrocytomas. AB - During the physiological process of PCD, the cell initiates a sequence of events culminating in the disintegration of the cell into small, membrane-bound apoptotic bodies. The intrinsic part of the PCD program arises from the mitochondria when it releases cytochrome c from the mitochondrial intermembrane space into the cytosol, forming the caspase-activating complex or apoptosome. The family of caspases is involved in the execution of genetically controlled PCD. Caspase-3 is expressed in normal and neoplastically transformed human cells and, like other caspases, is synthesized as an inactive, 32kDa proenzyme. Caspase-6 cleaves nuclear mitotic apparatus protein (NuMA) and mediates the shrinkage and fragmentation of cell nuclei. Caspase-8 is an initiation caspase that activates the caspase cascade during apoptosis, while caspase-9 is the initiator caspase in the caspase cascade in apoptotic normal and neoplastically transformed cells. During our immunocytochemical study, a sensitive, four-step, alkaline phosphatase conjugated antigen detection technique was employed. The results did in fact demonstrate the presence of high apoptotic activity within the cellular microenvironment of high-grade astrocytomas and glioblastomas. The observations identified cytoplasmic expression of caspase-3 and caspase-6 in more than 50 per cent of tumor cells, caspase-8 and caspase-9 in more than 10 per cent of tumor cells in high-grade anaplastic ASTR and glioblastoma. The immunocytochemical expression pattern in about 10 per cent of the tumor cells for caspase-3 and caspase-6 and about 1 to 5 per cent of the tumor cells for caspase-8 and caspase 9 demonstrated a translocation tendency from the cytoplasm to the cell nuclei in the apoptotic cells. This phenomenon may play an important role in these tumors' maintenance of immune privilege and evasion of immune attacks. We suggest that caspase-3, -6, -8 and -9 immunocytochemistry could have prognostic and immunotherapeutic significance in the treatment of these highly malignant glial tumors. PMID- 15523900 TI - Detection of 22q11.2 deletion among 139 patients with Di George/Velocardiofacial syndrome features. AB - Cytogenetic and FISH analysis was performed in 139 patients to detect the pathognomonic of Di George/ Velocardiofacial syndrome (DGS/VFCS) deletion 22q11.2. An abnormal karyotype was revealed in 2/139 cases (47, XXY and 46, XX, 2p+). A deletion was found in 17/139 (12.2%) patients (14 males/ 3 females), inherited in 3 (2 maternal and 1 paternal). Patients with 22q11.2 deletion exhibited facial dysmorphic features (82%), congenital heart defects (70%), immunological problems (47%), multiple congenital anomalies (64%), hypocalcemia (47%), mental retardation/learning difficulties (35%), cleft palate/velopharyngeal insufficiency (23.5%), seizures/hypotonia (23%) and growth retardation (12%). Among 56/139 patients with detailed available clinical data, the 22q11.2 deletion was confirmed in all cases with hypocalcemia and in over half of the cases with multiple congenital anomalies, immunological problems and hypotonia/seizures (70%, 60% and 57%, respectively). Genetic reevaluation of 39 patients without the 22q11.2 deletion contributed to the classification of 14 (37%) under different syndromes, emphasizing the need for stricter referral criteria. PMID- 15523901 TI - Sequential expression of inducible nitric oxide synthase and cyclooxygenase-2 during DMBA-induced hamster buccal pouch carcinogenesis. AB - BACKGROUND: Although it is known that iNOS and COX-2 are abundantly expressed in oral premalignant and malignant lesions, respectively, the interaction between iNOS and COX-2 has not been extensively studied. The purpose of this study was to examine the alteration of the iNOS and COX-2 expression level during hamster buccal pouch (HBP) carcinogenesis. MATERIALS AND METHODS: The expression of both iNOS and COX-2 on normal, dysplastic mucosa and squamous cell carcinoma (SCC) from different differentiation stages in 7, 12-dimethylbenz[a]anthracene (DMBA) induced HBP carcinogenesis was examined using immunohistochemical analysis. RESULTS: The mean values of both iNOS and COX-2 expression increased gradually from control to dysplastic lesions and more to invasive SCC. The highest mean expression was SCC. The differences between both iNOS and COX-2 expression in the normal and that in the dysplastic and carcinoma lesions were statistically significant. CONCLUSION: The results suggest that iNOS can enhance its ability to promote tumor growth in cooperation with COX-2. The expression of iNOS and COX-2 may be one of the factors that contribute to oral carcinogenesis. PMID- 15523902 TI - The presence of anti-p53 antibodies in sera from patients with oesophageal carcinoma: correlation to treatment, tumour volume and survival. AB - BACKGROUND: A correlation between mutations in the p53 gene and the presence of anti-p53 antibodies in sera has been reported. The aim of the present study was to analyse anti-p53 antibodies in sera from patients with oesophageal carcinoma and their implications for clinical outcome and survival PATIENTS AND METHODS: Between 1996 and 2002, patients treated for oesophageal carcinoma at the Department of Oncology, Uppsala University Hospital, Sweden, were asked to donate serum samples during treatment and follow-up. A total of 42 patients, with serum samples collected prior to therapy, were analysed for expressions of anti-p53 antibodies using a commercially available sandwich ELISA (Dianova, Hamburg, Germany). RESULTS: Anti-p53 antibodies did not correlate with investigated laboratory parameters. No correlation between anti-p53 antibodies and tumour volume was found (n=31; r=0.08;p=0.66). Anti-p53 antibodies as a continuous variable was not associated with survival (p = 0.42). Neither was the presence of anti-p53 antibodies (according to defined cut-off of 1.1, provided by the manufacturer) associated with survival (p = 0.99). CONCLUSION: The presence of anti-p53 antibodies correlated neither to tumour volume nor to clinical parameters. PMID- 15523903 TI - Glutamatergic system in bone physiology. AB - Bone is a highly innervated tissue consisting of nerve fibers, which contain many neurotransmitters including glutamate. Recently, a number of studies have identified functional glutamate receptors in osteoblasts and osteoclasts, implying that the glutamatergic system has an apparent regulatory role in bone physiology. This review outlines the evidence which suggest that the glutamatergic system regulates bone physiology. PMID- 15523904 TI - Use of annexin V for the identification of fetal cells in maternal circulation. AB - MACS with Annexin V-conjugated microbeads was used to isolate cells in apoptosis from the peripheral blood of 112 women at different weeks of gestation and from 15 women (60 samples) sequentially tested postpartum. FISH using X/Y probes was applied to quantitate fetal apoptotic cells. The mean apoptosis rate in the 16th 18th week was 6.5% and fetal cells constituted 5.1% of the apoptotic cell population. In the 26th-28th week it was 10.1%/7.5% and in the 37th-38th week 12.5%/9.9%, respectively. In samples obtained 30 min, 12h and 24h postpartum, the mean apoptosis was 25.1%, 12.5 and 6.1%, respectively and fetal cells constituted 14.8%, 2.1% and 0.16% of the apoptotic cells. Forty-eight h after delivery, apoptosis was 2.3% and no fetal cells were detected. Accurate estimation of the proportion of fetal cells undergoing apoptosis may facilitate the optimization of protocols for non-invasive prenatal diagnosis of chromosomal abnormalities. PMID- 15523905 TI - Transdermal therapeutic fentanyl-system (TTS-F). AB - Fentanyl, a surgical analgesic and general anaesthetic, is a lipophilic short acting synthetic opioid, having a selective potent effect on mu receptors. The transdermal therapeutic fentanyl-system (TTS-F) allows for a continued and sustained titratable amount of fentanyl to be delivered without the inconvenience of the typical 24-h administration of other analgesics. Although incidences of respiratory depression led to TTS-F being contraindicated for postoperative analgesia, it is currently undergoing Phase III trials for nociceptive, neuropathic and chronic moderate to severe pain in a variety of settings. It demonstrates a slow pharmacokinetic profile and incidences of breakthrough pain may still require rapid analgesia, for which intravenous and bolus administration of rapid acting opioids remain 'gold standard' However, TTS-F is finding uses for chronic pain of cancer origin where it offers a solution for step 3-pain (WHO) management on the WHO analgesic ladder. More recent data indicates that TTS-F is not only effective for neuropathic but also nociceptive non-cancer and cancer pain alike. This review presents an overview of the synthesis, delivery, pharmacokinetics, toxicity and clinical pharmacology of the transdermal delivery of fentanyl. PMID- 15523906 TI - Expression of JNK interacting protein JIP-1 is down-regulated in liver from mouse embryos with a disrupted insulin-like growth factor II gene. AB - JNK interacting Protein 1 (JIP-1) is a pivotal scaffolding protein in the JNK signalling pathway. Its expression pattern in murine tissue indicates that JIP-1 has a role in the regulation of different cellular events. By examining the JIP-1 expression in transgenic mice that were heterozygous for a functional insulin like growth factor II (igf-2) gene, it was possible to show that an abrogated igf 2 expression was followed by a decreased transcription of the JIP-1 gene. This pattern was consistent through different litters, which suggests that the up- or down-regulation of JIP-1 may well be part of the intracellular mediation of IGF II induced messages. PMID- 15523907 TI - Interleukin 4 increases the antibody response against Rubisco in mice. AB - The influence of interleukin 4 (IL-4) on antibody titer in serum and spleen culture supernatant in mice immunized with spinach (Spinacia oleracea L.) Rubisco was investigated. Therefore, we boosted one mouse additionally to the antigen with recombinant mouse IL-4. We found that the Rubisco-specific antibody titer in serum as well as in spleen cell culture supernatant was significantly enhanced in the IL-4 mouse. Most of the antibodies were of the IgG1 subclass. After hybridoma generation, Rubisco-specific antibodies were found in more than 95% of the wells tested compared to about 12% of the control mouse. PMID- 15523908 TI - The effects of phosphoinositide/calcium- or cyclic AMP-mediated signal transduction pathway inhibitors on the activation of rat peritoneal macrophages by acetylated low-density lipoprotein. AB - BACKGROUND: Macrophages that uptake modified lipoproteins are activated and may initially behave as endotoxin-stimulated macrophages. This study was undertaken in order to determine whether signal transduction pathways controlling endotoxin mediated activation may also influence the lipoprotein-mediated activation of macrophages. MATERIALS AND METHODS: Rat peritoneal macrophages were incubated for 16 hours with acetylated low-density lipoprotein and certain agents that modify the phosphoinositide/calcium- and cyclic AMP-mediated pathways, such as 2-[4 morpholinyl]-8-phenyl-1[4H]-benzopyran-4-one (LY-294002), autocamtide 2-related inhibitory peptide (AIP), N-(2-[p-bromocinnamylamino]-ethyl)-5 isoquinolinesulfonamide hydrochloride (H-89) and actinomycin D. The production of nitric oxide and the intracellular and extracellular activities of acid phosphatase were assayed. RESULTS: Macrophages incubated with acetylated low density lipoprotein showed an increased production of nitric oxide and intracellular acid phosphatase activity as compared to their controls. LY-294002, AIP and H-89 caused a significant decrease in nitric oxide production and intracellular acid phosphatase activity. Actinomycin D had similar effects. AIP and actinomycin also significantly increased extracellular acid phosphatase activity. CONCLUSION: The activation of peritoneal macrophages by acetylated low density lipoprotein was similar to the activation by endotoxin, as expressed by the nitric oxide production and acid phosphatase intracellular activity; agents controlling the phosphoinositide/calcium- and cyclic AMP-mediated pathways in endotoxin-activated macrophages also influence the acetylated low-density lipoprotein-activated macrophages. PMID- 15523909 TI - Expression of mismatch repair proteins in invasive and in situ carcinoma of the breast. AB - BACKGROUND: Genetic instability is a characteristic feature of familial and sporadic breast carcinomas. It is not clear whether defects in the mismatch repair system accompany this instability. The purpose of this study was to explore the expression of two of the proteins encoded by the DNA mismatch repair genes, namely MLH1 and MSH2, in sporadic in situ and invasive breast carcinomas of various types and grades occurring in Greek patients. MATERIALS AND METHODS: MLH1 and MSH2 expression was monitored immunohistochemically in 60 breast carcinomas (20 in situ and 40 invasive). RESULTS-CONCLUSION: Although we did not detect loss of MLH1 or MSH2 expression, we do believe that our data will contribute to a better understanding of the role of the mismatch repair (MMR) system in breast cancer. PMID- 15523910 TI - Evaluation of leptin, interleukin-1beta, tumor necrosis factor-alpha and vascular endothelial growth factor in serum and follicular fluids of women undergoing controlled ovarian hyperstimulation as prognostic markers of ICSI outcome. AB - BACKGROUND: Cytokines play an important but controversial role during ovarian folliculogenesis for the development of mature and fertilizable oocytes. In this study, leptin, interleukin-1beta (IL1beta), tumor necrosis factor-alpha (TNFalpha) and vascular endothelial growth factor (VEGF) in serum and follicular fluids (FF) of women undergoing ovarian hyperstimulation were evaluated as prognostic markers of the outcome of intracytoplasmic sperm injection (ICSI) cycles. MATERIALS AND METHODS: Ninety-five ICSI cycles were included in the study. The cytokines were measured in serum and FF samples with enzyme immunoassay methods. RESULTS: The cytokine concentrations in serum were not significantly correlated with the cytokine concentrations in FFs. Serum IL1beta was inversely-correlated with the number of retrieved oocytes. Serum TNFalpha was negatively-correlated with fertilization rate. In FFs, TNFalpha was positively correlated with leptin. Leptin and VEGF in FFs were negatively-associated with pregnancy outcome. CONCLUSION: Leptin and VEGF concentrations in FFs may serve as prognostic markers of success after ovarian hyperstimulation and ICSI. PMID- 15523912 TI - Isolation and growth characteristics of an EDTA-degrading member of the alpha subclass of Proteobacteria. AB - A Gram-negative, ethylenediaminetetraacetic acid (EDTA)-degrading bacterium (deposited at the German Culture Collection as strain DSM 9103) utilising EDTA as the only source of carbon, energy and nitrogen was isolated from a mixed EDTA degrading population that was originally enriched in a column system from a mixture of activated sludge and soil. Chemotaxonomic analysis of quinones, polar lipids and fatty acids allowed allocation of the isolate to the alpha-subclass of Proteobacteria. 16S rDNA sequencing and phylogenetic analysis revealed highest similarity to the Mesorhizobium genus followed by the Aminobacter genus. However, the EDTA-degrading strain apparently forms a new branch within the Phyllobacteriaceae/Mesorhizobia family. Growth of the strain was rather slow not only on EDTA (micro(max) = 0.05 h(-1)) but also on other substrates. Classical substrate utilisation testing in batch culture suggested a quite restricted carbon source spectrum with only lactate, glutamate, and complexing agents chemically related to EDTA (nitrilotriacetate, iminodiacetate and ethylenediaminedisuccinate) supporting growth. However, when EDTA-limited continuous cultures of strain DSM 9103 were pulsed with fumarate, succinate, glucose or acetate, these substrates were assimilated immediately. Apparently, the strain can use a broader spectrum than indicated by traditional substrate testing techniques. The EDTA species CaEDTA and MgEDTA served as growth substrates of the strain because in the mineral medium employed EDTA was predicted to be mainly present in the form of these two complexes. The bacterium was not able to degrade Fe3+-complexed EDTA. PMID- 15523911 TI - Effects of Bacillus subtilis O9 biosurfactant on the bioremediation of crude oil polluted soils. AB - The application of a surfactant from Bacillus subtilis O9 (Bs) on the bioremediation of soils polluted with crude oil was assayed in soil microcosms under laboratory conditions. Three concentrations of biosurfactant were assayed (1.9, 19.5, and 39 mg kg(-1) soil). Microcosms without biosurfactant were prepared as controls. During the experiment, the crude oil-degrading bacterial population, the aliphatic and aromatic hydrocarbons were monitored in each microcosm. The results indicated that applying Bs did not negatively affect the hydrocarbon-degrading microbial population Concentrations of 19 and 19.5mg (Bs) per kilogram of soil stimulated the growth of the population involved in the crude oil degradation, and accelerated the biodegradation of the aliphatic hydrocarbons. However, none of the assayed Bs concentrations stimulated aromatic hydrocarbon degradation. PMID- 15523913 TI - Biodegradation of dibenzo-p-dioxin, dibenzofuran, and chlorodibenzo-p-dioxins by Pseudomonas veronii PH-03. AB - The dioxin-degrading strain Pseudomonas veronii PH-03 was isolated from contaminated soil by selective enrichment techniques. Strain PH-03 grew on dibenzo-p-dioxin and dibenzofuran as a sole carbon source. Further, 1 chlorodibenzo-p-dioxin, 2-chlorodibenzo-p-dioxin and other dioxin metabolites, salicylic acid, and catechol were also metabolized well. Resting cells of strain PH-03 transformed dibenzo-p-dioxin, dibenzofuran, 2,2',3-trihydroxybiphenyl, and some chlorodioxins to their corresponding metabolic intermediates such as catechol, salicylic acid, 2-hydroxy-(2-hydroxyphenoxy)-6-oxo-2,4-hexadienoic acid, and chlorocatechols. The formation of these metabolites was confirmed by comparison of gas chromatography-mass spectrometry (GC-MS) data with those of authentic compounds. Although we did observe the production of 3,4,5,6 tetrachlorocatechol (3,4,5,6-TECC) from 1,2,3,4-tetrachlorodibenzo-p-dioxin (1,2,3,4-TCDD) with resting cell suspensions of PH-03, growth of strain PH-03 in the presence of 1,2,3,4-TCDD was poor. This result suggests that strain PH-03 is unable to utilize 3,4,5,6-TECC, even at very low concentration (0.01 mM) due to its toxicity. In cell-free extracts of DF-grown cells, 2,2',3-trihydroxybiphenyl dioxygenase, 2-hydroxy-6-oxo-6-phenyl-2,4-hexadienoic acid hydrolase, and catechol-2,3-dioxygense activities were detected. Moreover, the activities of meta-pyrocatechase and 2,2',3-trihydroxybiphenyl dioxygenase from the crude cell free extracts were inhibited by 3-chlorocatechol. However, no inhibition was observed in intact cells when 3-chlorocatechol was formed as intermediate. PMID- 15523914 TI - Effect of iron bioavailability on dissolved hydrogen concentrations during microbial iron reduction. AB - Dissolved hydrogen (H2) concentrations have been shown to correlate with specific terminal electron accepting processes (TEAPs) in aquifers. The research presented herein examined the effect of iron bioavailability on H2 concentrations during iron reduction in flow-through column experiments filled with soil obtained from the uncontaminated background area of the Field Research Center (FRC), Oak Ridge, TN and amended with acetate as the electron donor. The first column experiment measured H2 concentrations over 500 days of column operation that fluctuated within a substantial range around an average of 3.9 nM. Iron reduction was determined to be the dominant electron accepting process. AQDS (9,10 anthraquinone-2,6-disulfonic acid) was then used to determine if H2 concentrations during iron reduction were related to iron bioavailability. For this purpose, a 100-day flow-through column experiment was conducted that compared the effect of AQDS on iron reduction and subsequent H2 concentrations using two columns in parallel. Both columns were packed with FRC soil and inoculated with Geobacter sulfurreducens but only one was supplied with AQDS. The addition of AQDS increased the rate of iron reduction in the flow-through column and slightly decreased the steady-state H2 concentrations from an average of 4.0 nM for the column without AQDS to 2.0 nM for the column with AQDS. The results of this study therefore show that H2 can be used as an indicator to monitor rate and bioavailability changes during microbial iron reduction. PMID- 15523915 TI - Anaerobic degradability of alcohol ethoxylates and related non-ionic surfactants. AB - The anaerobic degradability of alcohol ethoxylates with various degrees of branching and several related substances was studied. Different inocula were employed in order to increase the probability of obtaining capable bacteria, and the degradation assays were fed with several small doses of the test substances in order to avoid inhibition by too high initial concentrations. Mineralization was quantified by monitoring the biogas production and inorganic carbon concentration in the liquid phase. Almost complete mineralization was achieved in the assays with linear alcohol ethoxylate, poly(ethylene glycol), dodecanol, 2 ethyl-hexanoic acid and 3-methyl-valeric acid. No significant degradation was detected in the assays with highly branched alcohol ethoxylate, 2-butyl-branched alcohol ethoxylate, alcohol alkoxylate, poly(propylene glycol) and iso tridecanol. A 2-ethyl-branched alcohol ethoxylate was transformed to (2-ethyl hexyloxy)-acetate, which was not further degraded. Apparently already the first step of anaerobic degradation of alcohol ethoxylates, the ethoxylate chain shortening, is sterically hindered by the alkyl branching. Alkyl branching in alcohol ethoxylates and the inclusion of propylene oxide units in alcohol alkoxylates seem to have a clearly more detrimental effect on anaerobic degradability than on aerobic degradability. PMID- 15523916 TI - Biotransformation and dissolution of petroleum hydrocarbons in natural flowing seawater at low temperature. AB - The objective of this study was to establish methods for controlled studies of hydrocarbon depletion from thin oil films in cold natural seawater, and to determine biotransformation in relation to other essential depletion processes. Mineral oil was immobilized on the surface of hydrophobic Fluortex fabrics and used for studies of microbial biodegradation in an experimental seawater flow through system at low temperatures (5.9-7.4 degrees C) during a test period of 42 days. The seawater was collected from a depth of 90 m, and microbial characterization by epifluorescence microscopy, fluorescence in situ hybridization, and most-probable number analysis showed relatively larger fractions of archaea and oil-degrading microbes than in the corresponding surface water. Chemical analysis of hydrocarbons attached to the fabrics during the test period showed that n-alkanes (C10-C36) were decreased by 98% after 21 days, while naphthalenes were depleted by 99-100% during the same period. At the end of the period 4-5 ring polyaromatic hydrocarbon (PAH) compounds were removed by 82% from the fabrics. Analysis of the recalcitrant pentacyclic triterpane C30 17alpha(H),21beta(H)-hopane showed that the oil remained adsorbed to the fabrics during the test period. Comparison of depletion analysis with calculation of hydrocarbon dissolution in a flow-through system indicated that naphthalenes and smaller PAH compounds (alkylated 2-ring and 3-ring compounds) were removed from the fabrics by dissolution. The data further implied that depletion of n-alkanes and 4-5 ring PAH hydrocarbons were the result of biotransformation processes. PCR amplification of bacterial 16S rRNA genes from microbes adhering on the immobilized oil surfaces showed the dominance of a few bands when analysed in denaturing gradient gel electrophoresis (DGGE). Sequence analysis of DGGE bands revealed phylogenetic affiliation to the alpha- and gamma-subdivisions of proteobacteria and to the Chloroflexus-Flavobacterium-Bacteroides group. PMID- 15523917 TI - Insights into amyloid structural formation and assembly through computational approaches. AB - Amyloids are long, insoluble ordered fibers. Due to their insolubility, to date the determination of an amyloid structure with an atomic scale resolution has proven to be a difficult task. Under such circumstances, computational approaches are a preferred option, providing the means to build likely models, test their stabilities and figure out the chemistry of their prevailing interactions. Computational models can be validated by targeted experiments, such as introducing mutations and testing for amyloid formation. Computations further provide vehicles for the comprehension of the mechanisms of amyloid seed formation and oligomer toxicity. Nevertheless, computations face an immense hurdle, the outcome of the time scales involved in amyloid formation and the immense sizes of the systems. In an attempt to overcome these, we adopt a strategy that encompasses (1) bioinformatics studies of native proteins containing beta-sheet structures; (2) simulations of shorter peptides; and finally (3) construction of potential oligomeric models and tests of their stabilities. The results are correlated with experimental data where available. Here, we describe the computational methods in simple terms and present an overview of the results. The systems derive from amyloidogenic, disease-related proteins, including gelsolin, beta2-microglobulin, and peptides derived from the prion, Alzheimer's Abeta, IAPP and human calcitonin. Ultimately, obtaining molecular structures should facilitate efforts to therapy and drug design. PMID- 15523918 TI - Folding and misfolding of the prion protein in the secretory pathway. AB - A hallmark of prion diseases in humans and animals is the conversion of the cellular prion protein PrPc to a pathogenic isoform, denoted PrPSc. PrPSc is characterized by distinct biochemical and biophysical properties; in addition, it is the major component of infectious prions. All available data indicate that the only difference between PrPc and PrPSc resides in their conformation, emphasizing a critical role of protein folding in the pathogenesis of prion diseases. PMID- 15523919 TI - Immunohistochemical detection of beta-amyloid and beta-amyloid precursor protein in the canine brain and non-neuronal epithelial tissues. AB - We examined the immunohistochemical localization of beta-amyloid (Abeta) and beta amyloid precursor protein (AFPP) in the neuronal and non-neuronal tissues of 14 aged (over 10 years old) and 4 young (0-4 years old) dogs. Abeta was detected only in the senile plaque in the cerebrum of aged dogs. AbetaPP was expressed in the neuronal cell body, neuronal fiber, senile plaque and perimalacic tissue independent of age. In addition, epithelial cells in the bronchus, bronchial glands, gastric and intestinal mucosa, intrahepatic bile ducts, and pancreatic ducts and exocrine glands were immunopositive for AbetaPP. Thus, it is suggested that AbetaPP may be expressed in the non-neuronal epithelial tissues independent of age and of the presence of senile plaques, and the expression may depend on individual differences or physiological conditions. PMID- 15523920 TI - Kappa III immunoglobulin light chain origin of localized orbital amyloidosis. AB - Isolated orbital amyloidosis is a rare condition in which intra-muscular deposits result in proptosis and restriction of eye movement. Previous reports have suggested an immunoglobulin origin of the amyloid fibrils, but this has not been proven biochemically. A case is presented in which initial unilateral orbital amyloidosis progressed to bilateral disease. Biochemical analysis of resected ocular muscle determined that the amyloid fibrils were derived from a kappa III immunoglobulin light chain. Questions of pathogenesis and tissue tropism are considered. PMID- 15523921 TI - Serum amyloid A protein forms a complex with a fragment of apolipoprotein A-I in the domestic blue fox: a protective mechanism against AA amyloidosis? AB - The spontaneous occurrence of protein AA-type of amyloidosis varies among animal species. As reactive AA-type of amyloidosis has never been detected in the blue fox, we obtained acute phase sera to search for amyloid-protective elements. The purified SAA fraction was characterized by mass and sequence analyses to disclose any unique domains in the amino acid sequence. The data revealed an SAA protein with heterogeneities in several positions, and showed the typical insertion between positions 69 and 70. By comparing the amino acid sequence with that from other mammals, no unique sequence could be observed. However, a C-terminal fragment of apolipoprotein A-I (ApoA-I) was found attached to the SAA. The amino acid sequence of the ApoA-I fragment revealed a partially blocked and ragged N terminus. A comparison of the amino acid sequence of ApoA-I with that from the dog showed that the ApoA-I fragment started about position 190, had an intact C terminus, and showed an identical sequence in all positions, except one. Based on the data, we suggest an interaction of the C-terminal fragment of ApoA-I with the SAA protein that inhibits the AA fibrillogenesis in the blue fox. PMID- 15523922 TI - Coexistence of familial transthyretin amyloidosis ATTR Val30Met and spinocerebellar ataxia type 1 in a Japanese family--a follow-up autopsy report. AB - Three brothers in a family with Val30Met transthyretin (TTR) amyloid polyneuropathy (FAP) in Iiyama, Japan were studied pathologically. In this family, affected members have been reported to show typical clinical features of FAP, and some have been documented to exhibit symptoms and signs of central nervous system (CNS) involvement consisting of cerebellar ataxia and pyramidal tract signs. After the original description, this family was regarded as a unique phenotype of this form of FAP; however, subsequent molecular genetic studies revealed that some patients and asymptomatic members in the family had Val30Met TTR and/or spinocerebellar ataxia type 1 (SCA1) gene mutations. In this study, pathological examination of two patients with both FAP and CNS symptoms showed (1) TTR-immunoreactive leptomeningeal and cerebrovascular amyloid deposition compatible with Val30Met TTR FAP, and (2) neuronal loss and gliosis mainly in the Purkinje cell layer, spinocerebellar system, olivo-ponto-cerebellar system, dentato-rubral system, gracile nuclei, cuneate nuclei, and various nuclei of cranial nerves, accompanied by anti-expanded polyglutamine tract antibody positive neuronal intranuclear inclusions, all of which were compatible with the pathological findings of SCA1. On the other hand, the remaining patient with FAP symptoms only showed the former pathological finding alone. The present study demonstrates, at the pathological level, that Val30Met TTR FAP and SCA1 coexist in the same family members, and that the CNS dysfunction seen in the patients in this family is ascribable to SCA1 pathology but not to CNS amyloidosis. PMID- 15523923 TI - Mutant fibrinogen A-alpha-chain associated with hereditary renal amyloidosis and peripheral neuropathy. AB - A middle age Portuguese woman was investigated for renal amyloidosis. She presented with progressive renal failure, proteinuria, hypertension, and sensory symptoms in the feet. Clinical and neurophysiological evaluation disclosed sensory-autonomic neuropathy. Cardiovascular tests and 123-MIBG investigation showed parasympathetic dysfunction and decrease of myocardial innervation, in accordance with small fiber neuropathy, as usually observed in amyloidosis. Immunohistochemical studies revealed AFib amyloidosis and genetic studies the amino acid exchange Glu526Val of the fibrinogen Aalpha-chain mutation, which was also present in one of her sons. The mutant gene in this patient was associated with the same haplotype as all other reported cases of Glu526Val mutations. This is the first reported AFibamyloidosis in Portugal, and the first case of AFib in which sensory and autonomic nerve fiber dysfunction is described, indicating that small nerve fiber lesion can occur in the fibrinogen Aalpha chain mutation. This can be important for prognosis, in particular when liver transplantation is considered for treatment. PMID- 15523924 TI - Outcome of exercise electrocardiography in familial amyloidotic polyneuropathy patients, Portuguese type, under evaluation for liver transplantation. AB - Familial amyloidotic polyneuropathy (FAP) is a dominantly inherited systemic amyloidosis caused by mutated transthyretin (TTR). Liver transplantation is currently the only available treatment that halts the progress of the disease. Cardiovascular complications are common in FAP, and cardiac arrhythmias are typical complications in FAP Val30Met. For patients with late onset FAP, as the Swedish patients, coronary heart disease has been found in several patients, and a QS complex is not an uncommon finding in FAP-patients ECG raising the suspicion of coronary heart disease. The aim of this study was to evaluate exercise ECG in FAP patients before transplantation with regard to mortality and morbidity. Thirty-eight FAP patients who underwent examination by exercise ECG, as part of the evaluation for liver transplantation were included in the study. Of these, 30 patients were transplanted, and the surviving patients were followed for at least 2 years. Exercise ECG was performed on bicycles with standard 12 leads. Non parametric statistical analyses were used in all calculations. Six patients died 0-5.5 years after transplantation. They were older than the survivors (p < 0.01), but their duration of disease did not deviate from that of survivors (p = 0.8). They were also less able to increase their heart rates during exercise than the survivors (p < 0.05). For all transplanted patients, a significant relationship was found between patients' increase of heart rate, blood pressure and maximal workload, and the duration of disease and also for the PND-score, signifying that the outcome of exercise ECG predominantly was related to the patients autonomic and motor function, and not to their heart function. PMID- 15523925 TI - [Between a psychiatric diagnosis and a familial diagnosis]. AB - If the idea of a biopsychosocial model is not going to be merely a phrase, then every clinical diagnosis and therapeutic vision need to consider the familial issues. Taking the family issues into consideration has evident gains; it allows for a better understanding of the patient and allows for actions which reduce the risk of a relapse of illness. The fact that such issues are not part of the therapeutic armament, despite the effectiveness of psychoeducation, is bewildering: on one hand the families are not always invited to co-operate, on the other hand--they often give up this co-operation. In such a situation, the relation between the patient's family and the personnel needs analysis, especially common, often unconscious emotions and prejudices. They can form a dysfunctional 'knot', which does not allow for a therapeutic alliance to be formed. How to come out from such an impasse and make a common positive alliance? Paradoxically, the therapeutic possibilities are bigger when--especially in the initial phase of treatment--the psychiatrist "is able to not know" and is capable of listening into the patient's family perspective, and has an interest in the 'family story'. It is only after the knowledge on the family is gained and the family perspective is accepted, that the deconstruction of that part of the family's story that forms the problem and a common search for "positive changes" are possible. Although it may not appear to be so, such an ordeal can be very difficult for the psychiatrist who is used to controlling the situation through psychiatric theory and diagnosis, psychiatric language and the psychiatric institution. The issue presented appears to be part of a universal dilemma: what is the range of "the language power" of the psychiatrist and when is the sharing of this power with the patient and the family beneficial for the therapy. PMID- 15523926 TI - [Schizotaxia--theoretical construct or a tool for clinical research?]. AB - From Kretschmer's trials binding personality traits with risk of psychoses have been described in literature. Still there is lack of one theory linking genetic factors with schizophrenia. In 1962 Meehl, introducing the term "schizotaxia", had been trying to find an answer to such a question. He described schizotaxia as subtle neuronal integration deficit caused by a single genetic factor, which depending on conditions, can give schizotypy or schizophrenia. Actually, this theory has only historical meaning. Recently Tsuang and Faraone reformulated the concept of schizotaxia used in clinical studies. Preliminary results lead to a conclusion that presence of schizotaxia has a detrimental influence on social functioning, which is improving after neuroleptic therapy. Studies confirmed that the risk of schizophrenia in persons with schizotaxia was higher as compared to persons without such characteristics. It is supposed, that paying attention to traits of schizotaxia will improve the possibility of early diagnosis of schizophrenia. PMID- 15523927 TI - [Core factors of schizophrenia structure based on PANSS and SAPS/SANS results. Discerning and head-to-head comparisson of PANSS and SASPS/SANS validity]. AB - AIM: A lot of inconsistencies across dimensional studies of schizophrenia(s) are being unveiled. These problems are strongly related to the methodological aspects of collecting data and specific statistical analyses. Psychiatrists have developed lots of psychopathological models derived from analytic studies based on SAPS/SANS (the Scale for the Assessment of Positive Symptoms/the Scale for the Assessment of Negative Symptoms) and PANSS (The Positive and Negative Syndrome Scale). The unique validation of parallel two independent factor models was performed--ascribed to the same illness and based on different diagnostic scales- to investigate indirect methodological causes of clinical discrepancies. METHOD: 100 newly admitted patients (mean age--33.5, 18-45, males--64, females--36, hospitalised on average 5.15 times) with paranoid schizophrenia (according to ICD 10) were scored and analysed using PANSS and SAPS/SANS during psychotic exacerbation. All patients were treated with neuroleptics of various kinds with 410mg equivalents of chlorpromazine (atypicals:typicals --> 41:59). Factor analyses were applied to basic results (with principal component analysis, normalised varimax rotation). Investing the cross-model validity, canonical analysis was applied. RESULTS: Models of schizophrenia varied from 3 to 5 factors. PANSS model included: positive, negative, disorganisation, cognitive and depressive components and SAPS/SANS model was dominated by positive, negative and disorganisation factors. The SAPS/SANS accounted for merely 48% of the PANSS common variances. The SAPS/SANS combined measurement preferentially (67% of canonical variance) targeted positive-negative dichotomy. Respectively, PANSS shared positive-negative phenomenology in 35% of its own variance. The general concept of five-dimensionality in paranoid schizophrenia looks clinically more heuristic and statistically more stabilised. PMID- 15523928 TI - [The location of the deficits of emotional communication in dimensional models of schizophrenia]. AB - Disordered communication of emotions belongs to the primary schizophrenia symptoms. The aim of this study was our attempt of the localisation of the deficits in emotional communication (its decoding and expression) in dimensional models of schizophrenia. The evaluation of emotionality in schizophrenia might be performed indirectly via negative dimension (for instance: apathy, diminished verbal fluency, flat or inappropriate emotional reactions), via excitement (elevated mood) and depression (decreased mood) or via cognitive dimension. Interpretation of emotional phenomena within the cognitive dimension remains an oversimplification. Brain emotional and cognitive systems act independently although they also reveal mutual interactions enabling the control of emotional behaviour via social learning of emotions. To conclude, to date emotional dimension in schizophrenia has not yet received satisfactory explanation which is in line with the complex nature of emotions. Thus the question regarding the location of emotions in dimensional models of schizophrenia still remains open. PMID- 15523929 TI - [Features of the self-structure and source monitoring disorders in people with schizophrenia]. AB - AIM: The aim of this study was to explore relationships between self-structure and source monitoring in people with schizophrenia. METHOD: Forty-one outpatients with an ICD-10 diagnosis of schizophrenic disorders participated in the study. Subjects were asked to select personality trait words from a checklist that described themselves, themselves as they were five years ago, and what most people are like. They also performed a source monitoring task and were assessed on current psychiatric symptoms. RESULTS: Patients who lacked a sense of continuity over time tended to attribute self-generated items to an external source. Subjects who had a less clearly defined self-structure and whose self representation was less differentiated from others-representation tended to falsely attribute items to themselves. CONCLUSION: The basic features of self structure were significantly related to some problems in discrimination of self generated stimuli in the source monitoring task. PMID- 15523931 TI - [Working memory dysfunction in patients suffering from schizophrenia and their first-degree relatives]. AB - Working memory deficits are considered to play an important role affecting not only the pathogenesis but also the course of schizophrenia. Numerous studies of schizophrenic patients and their first-degree relatives suggest that this impairment may be an indicator of susceptibility to developing schizophrenia. AIM: A comparison of selected working memory indicators in patients with schizophrenia, their first-degree relatives, and healthy controls. METHOD: Participants in the study were 99 patients with the diagnosis of schizophrenia (according to the ICD- 10-DCR criteria) in an early period of remission, their healthy first-degree relatives (N = 42), relatives with a history of psychiatric disorder (N = 14), and a control group of participants (N = 54) unrelated to the subjects and with no psychiatric history. Selected tests from the computer-aided Vienna Test Battery were used, measuring: reaction time (RT) in a task that required choosing among complex stimuli of two modalities, a tendency to perseveration (PERSEV), and immediate visuospatial memory span (CORSI). RESULTS: Schizophrenic patients' performance was found to be significantly inferior to that of controls on all the cognitive tests related to working memory: they had a longer reaction time in forced choice tasks, elevated perseverative tendencies and reduced immediate visuospatial memory span. Moreover, healthy relatives of schizophrenic patients performed significantly poorer than did the controls both as regards perseverative tendencies and visuospatial memory scan. Schizophrenic patients did not differ significantly from their close relatives in the degree of visuospatial memory span impairment. CONCLUSIONS: The findings indicate that working memory deficits as assessed by the tests used in the study may be related to familial susceptibility to schizophrenia. Therefore, the dysfunction may be taken into account in the capacity of endophenotype of such susceptibility. PMID- 15523930 TI - [P50 sensory gating disorders of auditory evoked potentials (AEP) in persons with schizophrenia]. AB - The paper presents a review of recent data on research and clinical significance of gating of the P50 component of the auditory evoked potentials (AEP). Information filters are a necessary element for the proper functioning of the brain. It appears as though they have an important role in the information transfer mechanisms. Neurophysiologically, they appear hypothetically in the sensory gating of the P50 component of the AEP. Schizophrenic patients and their first degree relatives do not have proper sensory gating of the P50 AEP. This suggests that there is a common biological base for these disorders. Some clinical aspects of the schizophrenic psychoses can be linked to this disordered gating. There are also notes which show the contrary. Currently we do not know whether the improper sensory gating of the P50 AEP is a trait endophenotypically linked to schizophrenia, or only something that partially explains the pathophysiology of the illness--especially since the described phenomena may be evoked in healthy persons. PMID- 15523932 TI - [Attention impairment in patients suffering from schizophrenia and their relatives of first-degree]. AB - Attention dysfunction as assessed by standardised tests has often been reported in schizophrenic patients. Since many cognitive dysfunctions noted in these patients occur also in their close relatives, a supposition arises that they may be indicators not so much of transient states, but rather of a stable feature of cognitive functioning. This feature perhaps is transmitted from generation to generation and might contribute to the onset of the disease. AIM: A comparison of selected attention indicators in schizophrenic patients and their first-degree relatives with these in healthy controls without family history of schizophrenia. METHOD: Participants in the study were 99 patients diagnosed with schizophrenia (according to the ICD-10-DCR criteria) in an early period of remission, their first-degree relatives (N = 56), out of whom 42 were healthy and 14 had a history of psychiatric disorders, and a control group of participants (N = 42) unrelated to the subjects and with no psychiatric history. Several tests of the computer aided Vienna Test Battery were used, and namely: the RT test measuring reaction time to simple visual or auditory stimuli, and LVT test measuring the accuracy and performance time in a task that consisted in visual tracking of lines, and required concentration of visual perception. RESULTS: As compared to the controls, the patient group was found to manifest attention deficits in the form of longer reaction time to simple stimuli as well as increased performance time and decreased correctness of visual line tracking. The patients had also a somewhat longer reaction time to visual (but not auditory) stimuli, and their line tracking was inferior as compared to that of their healthy relatives. As regards the reaction time to simple stimuli, the latter did not differ significantly from the controls, but in the line tracking test which required more attention, their performance was significantly inferior to that of the control group. PMID- 15523933 TI - [The paranoid syndrome in the course of a brain tumor--case study]. AB - An inpatient case of The Mental Hospital in Zurawica, in which an appearance of a brain tumor was preceded by a paranoid syndrome, was described in this article. One of the numerous clinical situations, when an appearance of a somatic disease was preceded by manifestation of psychic disturbances, is described in this article. PMID- 15523934 TI - [The endocrine mask of a developing process of schizophrenia--case study]. AB - In this research psychic and somatic symptoms related to disturbances of hypothalamus-hypophysis-peripheral regulation which may occur in the schizophrenic process were analysed. Authors discussed the problem of relations between hypothalamus neuroregulation and pathogenesis of endocrine disturbances which suggest the organic cause of obesity, hirsutism and secondary amenorrhea among women diagnosed with paranoid schizophrenia. Actual antipsychotic pharmacological treatment, including some side-effects: the metabolic (obesity) and the endocrine (hyperprolactinemia) ones were considered. The authors conclude that endocrine disorders which are connected with hypothalamus disfunction (sleeping, eating and reproductive functions) may reach the psychotic symptoms and treating them influences at the same time some endocrine changes. The estimation of PRL release in a test of stimulation with metoclopramide can be a sensitive (though not specific) test of dopaminergic activity in tuberous- infundibulum pathway and may be used to control the treatment. PMID- 15523935 TI - [Subjective and objective evaluation of olanzapine treatment in schizophrenia]. AB - AIM: The study was to evaluate objectively (by a physician) and subjectively (by the patients) the efficacy of 12-months olanzapine treatment of schizophrenic out patients. METHOD: Olanzapine treatment was either initiated or patients were switched from other antipsychotics to olanzapine. Patients' subjective evaluation with the use of SWN and DAI scales was compared with physicians' objective evaluation with the use of PANSS, CDS and CGI scales. RESULTS: The treatment with olanzapine caused significant improvement of schizophrenia positive, negative and affective symptoms. The use of the DAI questionnaire revealed also subjective improvement after olanzapine treatment. The decrease of schizophrenic symptoms correlated with the improvement of patients' subjective feeling of self-control, psychic functioning and somatic health. No serious adverse events were seen during the treatment. The severity of adverse events decreased during the olanzapine treatment. CONCLUSIONS: Olanzapine treatment of patients suffering from schizophrenia proved to be efficacious in both objective and subjective evaluation. PMID- 15523936 TI - [The influence of community support services on the costs of treatment of chronically mentally ill patients]. AB - AIM: To determine changes in costs of treatment for chronically mentally ill patients after their admittance to new social help units; Vocational Rehabilitation Center, Community Center of Mutual Help and Specialised Social Help Services at Client's Home. METHOD: In the group of 73 chronically mentally ill persons, for two years before and after their admission to social help units, the global amount of care (months x persons) and all the costs (PLN) in both social help and mental health systems were compared. RESULTS: For two years after the admission to the mentioned services all the costs in mental health system were significantly decreasing (71.9%), but new costs in social welfare system emerged. The overall costs of care were higher than before, but the elevation (35.4%) of all the costs was not so high as the increase (99,3%) of the care. The structure of care was improved (more day-, and less stationary care). The "out of the pocket" expenses for patients increased. CONCLUSIONS: The implementation of social help services for chronic mentally ill patients is reducing the amount of stationary and day treatment, but increasing active community treatment. Money in the health system is saved, but all the expenses on community care are increased. Coordination of care and finances between the mental health and social welfare is needed. Common financing for both systems is questionable because of impending over medicalisation. The allocation of money saved to early rehabilitation in schizophrenia would be profitable in the future. PMID- 15523937 TI - [Specifics of working with chronic mentally ill patients in Specialised Social Help Services--experiences in training and supervision meetings]. AB - Information was presented on the development and efficiency of The Specialised Psychiatric Social Help Services for the mentally ill in Poland. It took into consideration the specifics of this work and formal requirements for special training of the staff. The experiences of the 3.5 years of work of the specialised services in the Warszawa Targowek district (October 1997 - June 2001) were discussed. Organisational solutions and basic data about clients and staff and establishments and experiences coming from training and supervision meetings were discussed. Problems and difficulties most often experienced by the staff workers at their work with chronic mentally ill patients were described. Consideration was put from one side on the big rotation of the staff and from the other on the need of the extension of the theoretical and practical knowledge as well as of supervision and support was often mentioned in the questionnaire. A need of elaborating the programme of training and professional courses for workers of the specialised psychiatric social help services was also pointed out. PMID- 15523938 TI - [Parental and peer attachment of hospitalized adolescent patients--pilot study]. AB - AIM: In the context of Bowlby's theory of attachment authors made an attempt to assess parental and peer attachment in different clinical subgroups of hospitalised adolescents. METHOD: 142 patients--58 with the diagnosis of neurotic disorders, 49 eating disorders and 35 patients with schizophrenic disorders, as well 34 untreated, healthy adolescents were assessed by The Inventory of Parent and Peer Attachment by Armsden and Greenberg. RESULTS: The group of schizophrenic patients was characterised by disturbed attachment with peers and normal attachment with parents.Two other diagnostic subgroups, neurotic and eating disorders were similar as far as nature of attachment was concerned. The attachment was disturbed both with parents and peers, but the level of alienation in relations with peers in the group of eating disorder patients was similar to the level in the group of schizophrenic patients. This conclusion allows us to put the group of eating disorders closer to the group of schizophrenic, than neurotic patients, as far as attachment is concerned. CONCLUSIONS: The Inventory of Parent and Peer Attachment, used for the first time in Poland, can be seen as a promising instrument in assessing three dimensions of attachment: communication, trust and alienation in relations with parents and peers. PMID- 15523939 TI - [The concentrations of homocysteine, folic acid and vitamin B12 in alcohol dependent male patients]. AB - Metabolism of homocysteine (sulphur-containing amino acid) is accomplished in the remethylation cycle where vitamin B12 and folic acid are essential coenzymes. Markedly elevated homocysteine concentrations have been observed in patients with nutritional deficiencies of vitamin B12 and folate. Hyperhomocysteinemia in alcohol abusers may result from malnutrition and disorder of intestine absorption. AIM: The aim of the study was the estimation of homocysteine, folic acid and vitamin B12 concentrations in alcohol dependent male patients. METHOD: 71 males with a clinical diagnosis of alcohol dependence (ICD-10) have been examined. The investigated parameters have been determined in the blood serum, the homocysteine by means of immunochemical method, vitamin B12 and folic acid by means of immunoenzymatic assay. RESULTS: Serum homocysteine concentration was significantly higher and serum folic acid concentration was lower in alcohol dependent men than in controls. Mean concentrations of folic acid and vitamin B12 were significantly lower in patients with hyperhomocysteinemia than in men with normal homocysteine concentration. The highest correlation was indeed noticed between folate deficiency and the intensity of hyperhomocysteinemia. CONCLUSIONS: The development of hyperhomocysteinemia is associated with alcohol dependence that is also a probable cause of folate and vitamin B12 deficiency. PMID- 15523940 TI - Pro- and antioxidant functions of quinones in mammalian cells. AB - This article is a brief review of current knowledge concerning some basic concepts about the redox and addition chemistry of quinones, followd by a survey of current information regarding the biochemistry of quinones in mammalian cells and their pro- and antioxidant functions. In recent years it has been recognized that ubiquinone (Coenzyme Q), in addition to its involvement as an electron and proton carrier in mitochondrial and bacterial respiration, acts in its reduced form (ubiquinol) as an antioxidant. The antioxidant activity, together with its high degree of hydrophobicity and its widespread occurrence in biological membranes and in low-density lipoprotein, suggest un important role of ubiquinol in cellular defense against oxidative damage. Degenerative diseases and aging may be manifestations of a decreased capacity to maintain adequate ubiquinol levels. PMID- 15523941 TI - Intra-arterial segmental thrombolysis in patients with arterial embolism of cardiac origin. AB - OBJECTIVE: It is well known that in the evolution of cardiac diseases, thromboembolic accidents are quite frequent. In patients with acute peripheral ischaemia, surgical treatment is usually the procedure of choice, but in some circumstances some of these patients cannot benefit from the surgical treatment. Conservative treatment rarely leads to satisfactory results. An alternative of therapy is the thrombolysis, but administration of Streptokinase in systemic dose can lead to serious thromboembolic accidents and hemorrhage. Intra-arterial administration of the drug, in small doses, can eliminate this risk. The aim of this study was to demonstrate the efficacy of intra-arterial thrombolysis in patients with cardiac diseases complicated with peripheral arterial emboli. METHOD: In the period 1984-1997, we performed a number of 84 intra-arterial thrombolyses. Sixty-one of them were performed in case of chronic or acute arterial obstruction of the inferior limbs, occurring in the evolution of chronic arterial disease, 14 in the patients with acute obstruction caused by emboli from cardiac cavities, and 9 in the patients with arterial embolism of other origin. We applied this procedure infusing Streptokinase intraarterially, continuously, directly and progressively in the thrombus, with a rate of 2-4000 unit/min, and a total dose more than 100,000-120,000 units. All patients had been consulted previously in a surgical unit and for different reasons surgical treatment was not indicated. RESULTS: We succeeded to repermeabilise the entire arterial route at all the patients (fact demonstrated by the clinical and Doppler examinations). We obtained complete primary repermeabilisation in 79% cases (11 cases), and repermeabilisation at 24 hours in all cases. We did not record major complications during this procedure. CONCLUSIONS: At the cases we studied (a small number), this method seems to be an alternative of therapy with good results in the restoration of obstructed arterial circulation. Intra-arterial administration of Streptokinase, in small doses, can eliminate the haemorrhagic and thromboembolic risks that appear in case of systemic administration of the drug. PMID- 15523942 TI - The value of intermittent inotropic therapy in unhospitalized patients with refractory heart failure. AB - The treatment of intractable heart failure (New York Heart Association--NYHA IV) with inotropic support is a well established adjunct in the control of clinical status of end-stage heart failure patients but usually needs many admissions and a long period of hospitalization. In this study we report on the follow-up findings of 64 patients for 18 months who were seen in an outpatients congestive heart failure program. All patients had their full standard oral drug treatment, received intensive patient teaching and weekly inotropic infusions. We assess the effects of this comprehensive therapeutic approach on: 1) the number of hospital admissions, 2) length of stay and 3) the number of emergency room visits during the ensuing year. These data were compared with similar data from the year before entry in the program for each patient. All our patients showed NYHA class IV heart failure and received dobutamine to manage their chronic heart failure in an outpatient setting. The cause of heart failure was ischaemic in 34 (53.12%), idiopathic in 14 (21.87%), hypertension in 10 (15.62%), pulmonary hypertension in 7 (10.93%) and valvular heart disease in 6 (9.37%) patients. The mean period of observation was 382 days. For the period before entry in the program patients had 67 emergency room visits, 184 admissions, and 832 days spent in the hospital. After enrollment all these figures significantly decreased (p<0.001), patients heading 35 emergency room visits, 112 admissions and 518 days spent in the hospital. In conclusion, the intermittent inotropic therapy as an outpatient setting may be a valuable cost-effective therapeutic method, in patients with refractory heart failure and is associated with the reduction of emergency room visits, admissions and days spent in the hospital. PMID- 15523943 TI - Results of Sotalol (Darob) therapy in ventricular arrhythmias. AB - The aim of this prospective study was to evaluate the benefit of Sotalol (Darob) use in the therapy of potential malignant ventricular arrhythmias and in the prophylaxis of potential malignant and malignant arrhythmias; the influence on mortality has also been considered. Eighty four patients (pts) diagnosed with ventricular extrasystoles (68 pts., namely 80.95%), ventricular tachycardia (9 pts., namely 10.71%) and ventricular fibrillation (7 pts., namely 8.34%) were included in this study. They have been monitored clinically, electrocardiographically and echocardiographically at 1,3,6 and 12 months (1 year). The drug was given orally in doses 80-320 mg/day in patients with ventricular extrasystoles and in mean doses of 160 mg/day, for 1 year, to prevent recurrencies of ventricular arrhythmias. CONCLUSIONS: 1. In malignant and potential malignant ventricular arrhythmias Darob has been efficient on long term (1 year) in 77.67% cases, while in curative treatment of non-malignant ventricular arrhythmias Darob has been efficient in 68.31% cases. 2. Only minor proarrhythmogenic effects have been noticed for the doses of Darob used in this study. 3. long-term administration of Darob seems to reduce the incidence of cardiovascular deaths (35.30% cardiac deaths and 64.70% non-cardiac deaths have been recorded; p < 0.05). 4. Darob constitutes a drug of choice in postinfarction ventricular arrhythmias therapy, due to the beta-blocking effect, favourable during the ischemic episodes which may trigger arrhythmias. PMID- 15523944 TI - The relationship between the lipid structure of the hair and acute myocardial infarction. AB - In a group of patients with acute myocardial infarction (AMI) an evaluation of the derangements in lipid metabolism was carried out by analysing the fat structure of the hair (closely reflecting serum lipid variations). The data obtained reflects the real incidence of dyslipidemia associated with myocardial infarction. Through hair analysis, the evaluation in time (over weeks, months) of lipids metabolism under hypolipidemic therapy can be carried out without taking repeated samples. PMID- 15523945 TI - The significance of chronic hepatitis B and C virus infections in some connective tissue diseases: the association with chronic liver disease. AB - BACKGROUND/AIM: We investigated the pathogenic role of chronic hepatitis B virus (HBV) and C virus (HCV) in some connective tissue diseases including systemic lupus erythematosus (SLE), overlap syndrome, rheumatoid arthritis (RA), seronegative spondylarthritis (SS) and the association with chronic liver disease. METHODOLOGY: There were studied 155 patients, aged among 18 to 64 years old: 57 with SLE, 22 with overlap, 26 with RA, 30 with SS. The diagnoses were established using modified ARA criteria. There were performed complex immunology tests, percutaneous liver biopsy, HLA, Elisa tests with Riba confirmation for detecting HCV and HBV. RESULTS: 17% of SLE patients were infected with hepatitis viruses, predominantly B (70%). Half of them had a hepatic involvement due to the hepatitis viruses. 23% of RA patients were equally B?C infected with only one case of hepatic involvement secondary to hepatitis viruses. All the HCV infected patients had rheumatoid factor (RF) IgG-IgM type, with low serum levels of haemolytic complement (CH50), increases serum levels of circulating immune complexes (CIC) and evidence of HLA DR4 In the group of SS 40% of patients were infected mostly with HBV. In HLA B27 (+) anchylosing spondylitis (AS) the incidence of HBV was 100%. CONCLUSIONS: There is no high prevalence of HCV infection in SLE, overlap syndrome or RA, compared to the control group. In SS the prevalence is increased (40%), especially HLA B27 (+) AS group (33%), in which HBV is noticed at a rate of 100%. The association SLE-HCV favours the visceral involvement especially renal ones, while the presence of HBV is associated with decrease of lupus activity. In RA, HCV induces IgG-IgM RF with complement activation, being considered as a trigger of the disease in HLA DR4 patients. In HLA B27 (+) AS. HBV may trigger the development of disease in genetically susceptible individuals. PMID- 15523946 TI - Interferon prophylaxis of hepatic carcinoma. AB - The present article reveals the importance of hepatic carcinoma among the other diseases in digestive oncology, and also the importance of a correct designation of these cases. Epidemiology and actual hypothesis on the mechanisms of oncogenesis are discussed. There are reviewed some studies in the literature concerning infection with hepatitis B virus, hepatitis C virus, coinfection (B and C viruses, B and D viruses), the role of interferon prophylaxis in such cases. Also there is present a statistics on chronic viral hepatits, cirrhosis of viral etiology and hepatic carcinoma, diagnosed in patients in "N.Gh.Lupu" Hospital, over two decades. PMID- 15523947 TI - Apolipoproteins A1 and B levels and serum cholinesterase activity in hyperlipidemic subjects. AB - Serum apolipoprotein B (apo B) levels were found to be significantly (p < 0.001) higher in the 27 patients with combined hyperlipidemia (144 m./dl +/- 27.6) than in the 17 normal weight normolipidemic control subjects (92 mg/dl +/- 20.6; X +/- SD). When compared to apolipoprotein A1 (apo A1) levels obtained in controls (168.5 mg/dl +/- 28.4), hyperlipidemic subjects displayed a moderate yet significant (p < 0.02) decrease of this apolipoprotein (140 mg/dl +/- 24.2). Serum apo B levels were significantly (p < 0.001) correlated with serum cholesterol concentrations and also, to a lesser degree (p < 0.01), with serum cholinesterase activity. A highly significant correlation (p < 0.001) between apo A1 and HDL cholesterol levels was also noted. The decrease ofHDL cholesterol occurring in hyperlipidemic men (-30%) was however more accentuated than the decrease of apo A1 (-18%) suggesting an enhanced transfer of cholesterol esters from HDL to VLDL and LDL. It is considered that the determination of apolipoproteins may be useful not only for the detection of risk factors for atherosclerosis, but also for a better insight concerning the mechanisms involved in the development of an atherogenic dyslipidemia. PMID- 15523948 TI - Flow cytometry measurements of intracellular Ca2+ in K 562 cells. AB - The aim of this study is to show the intracellular Ca2+ changes induced by methotrexate, a cell cycle inhibitor, in order to prove the intracellular Ca2+ implication in cell proliferation processes. The fluorescent dye used to measure this parameter is Fluo-3, a fluorescein derivative useful for measuring the kinetics of Ca2+ transitions, waves and oscillations. The alterations of Ca2+ concentration proper to K562 cells have been studied, using methotrexate as a cell activation factor, both as direct effects on the cell response (1-15 minutes), and effects in time (24h.), using methotrexate in the culture medium. The K562 response at Ca2+ level in time remained constant for the cells grown in methotrexate medium, showing a slight increase in the control medium. We proved that methotrexate, a cell cycle inhibitor decreased the cytoplasmatic Ca2+ concentration in K562 cells, soon after adding methotrexate both to T0 and T1. PMID- 15523949 TI - Magnesium orotate in myocardial and neuronal protection. AB - The nervous tissue and the myocardium have in common many denominators, such as: the inability to renew to substitute the severely damaged or dead cells, the role of the membrane electric activity, the presence of similar systems for antioxidation protection, which are obviously involved in pathologic events a.s.o. Aggressive factors mainly act by free radicals injury and increase in cytosolic calcium level. Magnesium orotate molecule includes two synergic protective components: orotic acid and magnesium. Moreover, the orotic acid behaves as a transporter, carrying magnesium into the cells. The antioxidation protective effect of the orotic acid is mainly due to the pirimidinic bases that favor and increased synthesis of enzymes which act as free radical scavengers. The cell antioxidation protective system is dramatically impaired following heavy aggressions such as the ischemia-reperfusion process. Magnesium orotate improves the survival of cells situated within the perinecrotic areas as well as of the cells secondarily damaged during the so-called "second wind". PMID- 15523950 TI - On what information do ophthalmologists base their clinical practice? PMID- 15523951 TI - Comparison of corneal flap thickness between primary and fellow eyes using three microkeratomes. AB - PURPOSE: To compare corneal flap thickness created in laser in situ keratomileusis (LASIK) in primary (right) and fellow (left) eyes (same blade for both eyes) using three microkeratomes. METHODS: The corneal thickness of 132 eyes (66 patients) was measured preoperatively and intraoperatively after flap creation. Corneal flap thickness was calculated by subtracting stromal bed thickness from total corneal thickness. Three microkeratomes were used: Nidek MK 2000, Bausch and Lomb Surgical Hansatome, and the Chiron Automated Corneal Shaper (ACS). Each patient had both corneas cut by one microkeratome and one blade at the same session. RESULTS: Mean corneal flap thickness created in primary eyes was 128.30 +/- 12.57 microm (range 105 to 147 microm) for the ACS (160-microm plate and 8.5-mm ring) and 122.96 +/- 13.30 microm (range 86 to 140 microm) for fellow eyes; Hansatome (160-microm plate and 8.5-mm ring): 141.16 +/- 20.11 microm (range 101 to 169 microm) in primary eyes and 120.95 +/- 26.95 microm (range 107 to 151 microm) in fellow eyes; Nidek (130-microm plate and 8.5-mm ring): 127.25 +/- 4.12 microm (range 116 to 134 microm) in primary eyes and 127.54 +/- 3.7 microm (range 119 to 134 microm) in fellow eyes. The corneal flap in the ACS and Hansatome microkeratomes was always thicker in the primary than the fellow eye, using the same blade for both eyes. No significant difference was found using the Nidek microkeratome. CONCLUSION: Corneal flap thickness tended to be thinner in fellow eyes than in primary eyes for the ACS and Hanstome microkeratomes. The Nidek microkeratome results were closer to specified corneal flap thickness than the ACS and Hanstome microkeratomes. PMID- 15523952 TI - Prospective randomized comparison of wavefront-guided and conventional photorefractive keratectomy for myopia with the meditec MEL 70 laser. AB - PURPOSE: To study refractive results and aberrometric changes in myopic patients treated with wavefront-guided photorefractive keratectomy (PRK) in comparison with standard PRK. METHODS: Sixty eyes of 60 patients with myopic astigmatism were randomly divided into two groups. Group 1 included 30 eyes (mean spherical equivalent refraction -4.39 +/- 1.31 D; range -2.50 to -6.50 D) treated with wavefront-guided PRK using the WASCA workstation and the Asclepion Meditec flying spot MEL 70 excimer laser. Group 2 had 30 eyes (mean spherical equivalent refraction -4.33 +/- 1.22 D; range -2.50 to -6.50 D) that underwent conventional PRK using the same laser, and served as the control group. Wavefront analysis of high order aberrations was performed before and 6 months after surgery. RESULTS: Postoperatively, wavefront error increased in both groups (5.0-mm wavefront aperture diameter). Six months after surgery, the eyes that received the WASCA ablation had a smaller increase in root-mean-square (RMS; 70% of increment) compared to the conventional PRK group (139% of increment) (P<.001). In the standard PRK group, all aberrations notably increased; in the wavefront-guided PRK group there was a smaller increase of trefoil and spherical aberrations (P<.001) and a decrease of coma aberrations (P<.001). The smaller increase of wavefront error in the wavefront-guided PRK group compared to the standard PRK group was more evident when preoperative RMS values were higher than 0.4 microm (P<.01). The visual parameters (spherical equivalent refraction, uncorrected and best spectacle-corrected visual acuity) did not show significant differences between the two groups. CONCLUSION: Wavefront-guided PRK induced a smaller increase of postoperative wavefront-error compared to conventional PRK, particularly in patients with higher preoperative higher order aberrations. PMID- 15523953 TI - Bilateral comparison of wavefront-guided versus conventional laser in situ keratomileusis with Bausch and Lomb Zyoptix. AB - PURPOSE: One aim of corneal refractive surgery is to correct defocus and astigmatism. In the process of correcting lower order aberrations, higher order ocular aberrations increase. To evaluate the effectiveness of wavefront-guided laser in situ keratomileusis (LASIK) in reducing the increase of higher order aberration, we compared aberrational change after LASIK with conventional and wavefront-guided customized ablation. METHODS: Our study included 48 eyes of 24 patients. We performed conventional LASIK in one eye (Group 1) and wavefront guided customized ablation in the other eye (Group 2). Ocular aberration was measured with the Zywave, a type of Shack-Hartmann aberrometer. We then compared low and high order aberrations, contrast sensitivity, visual acuity, corneal topography, and manifest refraction preoperatively and postoperatively at 1 and 3 months. RESULTS: Uncorrected visual acuity improved to more than 20/20 in two eyes in the conventional ablation group and in five eyes in the customized ablation group. In the conventional ablation group, Root-mean-square for higher order (RMS(H)) was 0.215 preoperatively, 0.465 (216.3%) at 1 month, and 0.418 (194.4%) at 3 months. In the customized ablation group, RMS(H) was 0.207 preoperatively, 0.380 (183.6%) at 1 month, and 0.371 (179.2%) at 3 months after LASIK. Mesopic contrast sensitivity in the customized ablation group was higher than that in the conventional ablation group, but this change was not statistically significant. CONCLUSIONS: Wavefront-guided customized ablation reduced the increase of high order aberrations resulting from LASIK. In terms of visual acuity, patient preference, and mesopic contrast sensitivity, wavefront guided customized ablation produced slightly-but not statistically significant better results. PMID- 15523954 TI - Wavefront-guided ablation with Bausch and Lomb Zyoptix for retreatments after laser in situ keratomileusis for myopia. AB - PURPOSE: To evaluate the efficacy of wavefront-guided ablation for treatment of residual refractive error and higher order aberrations after laser in situ keratomileusis (LASIK) for myopia. METHODS: Twenty-one eyes of 13 patients with residual refractive error after myopic LASIK were treated with a wavefront-guided ablation with the Zyoptix technique. In 13 eyes of 8 patients (Group A) we lifted the flap with an epitheliorhexis technique; in 8 eyes of 5 patients (Group B) we cut a new flap with a Hansatome microkeratome. RESULTS: Preoperative root mean square (RMS) values were 1.370 for second order aberrations, 0.382 for total higher order aberrations, 0.273 for third order, 0.243 for fourth order, and 0.052 for fifth order aberrations. Three months postoperatively, RMS values decreased significantly to 0.278 (second order), 0.189 (total higher order), 0.138 (third order), 0.107 (fourth order), and 0.038 (fifth order). Ablation depth was two times greater than needed with a standard Planoscan treatment. Patients in the microkeratome recut group showed a trend toward overcorrection, and residual astigmatism was higher (-0.75 +/- 0.58 D) than in the flap lift group (-0.25 +/- 0.29 D). CONCLUSIONS: Wavefront-guided ablation was an effective method to correct residual refractive error and higher order aberrations after myopic LASIK. Functional and refractive outcomes were better when we lifted the flap than when we recut the flap. PMID- 15523955 TI - One-year results of photorefractive keratectomy with and without surface smoothing using the technolas 217C laser. AB - PURPOSE: To assess the efficacy, predictability, stability, and safety of a smoothing technique in patients with myopia immediately after photorefractive keratectomy (PRK) using a scanning-spot excimer laser. METHODS: Using the Technolas 217C excimer laser, PRK was performed on 100 eyes of 54 patients. Ablation zone diameter was 6.0 mm and transition zone diameter was 9.0 mm. The eyes were randomized into two groups: in 50 eyes PRK alone was performed and in the other 50 eyes, a smoothing technique was performed after the initial ablation. Preoperative mean spherical equivalent refraction was -4.98 +/- 1.71 D in the PRK only group (range -2.25 to -8.60 D) and -4.82 +/- 1.61 D in the smoothing group (range -2.00 to -8.00 D). Follow-up was 12 months for all patients. RESULTS: At 1 year after surgery, mean manifest spherical equivalent refraction was -0.61 +/- 0.50 D (range -2.25 to +0.62 D) in the PRK only group and in the smoothing group, +0.02 +/- 0.32 D (range -0.75 to +0.75 D). Postoperative regularity topographic indices were lower in the smoothing group than in the PRK group (P<.001). CONCLUSIONS: Smoothing after PRK for correction of myopia up to -6.50 D increased surface regularity, as expressed by lower topography surface regularity indices, and reduced the incidence and severity of postoperative haze. We observed higher predictability throughout follow-up in the smoothing group, which may be addressed by a nomogram adjustment in the PRK only group. PMID- 15523956 TI - Smoothing of the ablated porcine anterior corneal surface using the Technolas Keracor 217C and Nidek EC-5000 excimer lasers. AB - PURPOSE: To demonstrate efficacy of a smoothing technique to increase regularity of the anterior corneal surface after photorefractive keratectomy (PRK), using two different excimer lasers. METHODS: Spherical ablations of -10.00 D were performed on 11 fresh porcine corneas using either the Technolas Keracor 217C scanning-spot or the Nidek EC-5000 scanning-slit beam excimer laser. Following the procedure, we performed a phototherapeutic keratectomy treatment (smoothing technique) on half of the corneal surface. The smoothing technique was performed using a viscous solution of 0.25% sodium hyaluronate, which was spread on the cornea prior to the procedure. The ablation zone was 6 mm in diameter and the transition zone extended to 3 mm. The ablation depth was set at 10 microm. Corneas were then examined with scanning electron microscopy. RESULTS: Smoother treatment zones were apparent in porcine corneas in which smoothing was performed following PRK, with both laser systems. Results from the two lasers were not directly compared. CONCLUSIONS: The smoothing procedure performed following PRK using a viscous 0.25% sodium hyaluronate masking solution and a scanning laser system rendered the porcine corneal surface more regular. PMID- 15523957 TI - Very high frequency ultrasound biometry of the anterior and posterior chamber diameter. AB - PURPOSE: To measure the largest diameter of the anterior chamber (AC) and posterior chamber (PC) dimension and its orientation and determine the relationship with the principal keratometric meridians. METHODS: Twenty-eight eyes of 14 subjects were scanned with high frequency (50 MHz) ultrasound in sequential meridional scan planes at 30 degrees increments. Observer identified angle and ciliary sulcus recess boundaries in each patient scan set were fit with an elliptical model to obtain the ellipse semi-major axis corresponding to the largest diameter and its meridional orientation. Anterior and posterior chamber diameters from raw data and model fit were compared using linear statistics. Circular statistics were used to compare the orientation of the largest diameter for raw ultrasound measurements, model estimations of largest diameter, and autorefractor determined keratometric axes. RESULTS: The mean model diameters were anterior chamber OD 12.07 mm (0.32 SD); anterior chamber OS 12.06 mm (0.36 SD); posterior chamber OD 12.35 mm (0.42 SD); posterior chamber OS 12.33 mm (0.43 SD). The general trend for orientation of the meridian of largest diameter was in the horizontal meridian. In over 35% of eyes the difference between AC or PC meridian and the flat keratometric axis was greater than 20 degrees. CONCLUSIONS: Accurate and reproducible anterior segment biometry depends on visualization of structures and minimization of eye and head movement error. The range and standard deviation of the diameter and orientation measures suggests anatomic variation is sufficient to require biometry for proper sizing and placement of intraocular devices that use angle or sulcus fixation. PMID- 15523958 TI - New formula to calculate corneal power after refractive surgery. AB - PURPOSE: To assess the validity of intraocular lens (IOL) power calculations utilizing a theoretical variable refractive index correlated to axial length after myopic photorefractive keratectomy (PRK) in a clinical simulation and in patients who underwent cataract surgery after PRK for myopia. METHODS: Our study included 374 eyes of 300 patients who had PRK for myopia (-2.00 to -12.00 D, mean -4.83 +/- 2.57 D), divided into three groups: Group I had 44 eyes with small ablation zones of 5 to 5.5 mm; Group II had 49 eyes with large ablation zones of 6 to 7 mm; Group III was the control group of 281 eyes (201 patients; 87 males and 114 females) with small and large ablation zones. PRK was performed using the Aesculap-Meditec MEL 60/94 and MEL 70 lasers, and the corneal power was acquired by corneal topography (EyeSys 2000) and a Nidek KM-800 keratometer. RESULTS: There was a higher correlation between corneal power and both the change in refraction and axial length when calculated using keratometric measurements. IOL power calculated using keratometric postoperative PRK power was underestimated. The difference between the mean calculated and actual IOL power for emmetropia was 4.30 +/- 2.34 D. A theoretical variable refractive index (obtained from eyes treated with large PRK ablation zones) that correlated with axial length provided the correct keratometric postoperative PRK power: difference between mean calculated and mean actual IOL power was 0.42 +/- 1.23 D. CONCLUSIONS: We propose a theoretical variable refractive index that is correlated to axial length. Utilizing this keratometric correct power, we calculated IOL power similar to that for emmetropia. PMID- 15523959 TI - Astigmatic power calculations for intraocular lenses in the phakic and aphakic eye. AB - PURPOSE: To develop sets of equations employed in the power calculations for toric intraocular lenses (IOLs) in phakic or aphakic astigmatic eyes. METHODS: Mathematical operations to convert from standard toric parameters of sphere, cylinder, and axis to astigmatic decomposition components, and vice versa, are presented. These operations are used to derive equations to calculate the ideal toric IOL power for a phakic or aphakic astigmatic eye, predict the postoperative spectacle correction for a selected toric IOL with power other than the ideal power, and back calculate a parameter to be used to optimize predictability of the calculations based on clinical data. RESULTS: Two numerical examples are provided to show how the equations are used with clinical data. CONCLUSION: The equations developed provide a method to perform toric IOL power calculations and supporting operations of predicted postoperative spectacle refraction and optimization of prediction error for phakic and aphakic eyes with astigmatism. PMID- 15523960 TI - Correlation of changes in refraction and corneal topography after photorefractive keratectomy. AB - PURPOSE: To establish which corneal power evaluation measured with corneal topography correlates best with refractive changes after photorefractive keratectomy (PRK) for myopia. METHODS: Two hundred fifty-one consecutive eyes of 171 patients who had PRK for myopia ranging from -14.80 to -0.50 D (mean -5.43 +/ 2.978 D), calculated at the corneal plane, were included in the analysis. Data included preoperative and postoperative (1, 3, and 6-mo) subjective refraction and videokeratography with a Keratron Scout (Optikon 2000). Statistical analysis was performed to determine the correlation between the change in subjective refraction at the corneal plane and changes in six corneal power measurements: best fit sphere, simulated keratometry (Sim K), corneal apex, and center of the pupil (last two evaluated for axial and meridional curvatures). RESULTS: The closest correlation between subjective refraction change and corneal power measurement during the three follow-up evaluations was found with Sim K (R2 = 0.904; 0.889; 0.854) and best fit sphere (R2 = 0.919; 0.909; 0.872), whereas the other measurements showed poor correlation with the different curvatures. CONCLUSIONS: The best fit sphere corneal topography parameter correlated best with the refractive changes, primarily for low treatment amounts, whereas it showed a clear-cut underestimation in eyes that had undergone high dioptric treatments. PMID- 15523961 TI - Normal values for photopic and mesopic letter contrast sensitivity. AB - PURPOSE: The exponential increase of patients having refractive surgery has increased the number of patients with night vision disturbances, such as decreased contrast sensitivity. However, there are no standard contrast sensitivity scales in normal persons in the mesopic range. We describe Pelli Robson contrast sensitivity under photopic and mesopic luminance conditions in a large Spanish population over a wide range of age groups to provide normal values. A further aim was to evaluate the effect of photopic visual acuity on photopic and mesopic contrast sensitivity. METHODS: A cross-sectional study was performed on 292 participants stratified by age into six groups. Binocular contrast sensitivity was determined with best spectacle correction using the Pelli-Robson letter chart at 1 m under photopic (85 cd/m2) and mesopic (0.15 cd/m2) luminance conditions. RESULTS: Phototopic letter contrast sensitivity began to decrease gradually from the 61 to 70-year-old age group onward, and for mesopic conditions, from the 51 to 60-year-old age group onward. The reduction in mean contrast sensitivity between the oldest and the youngest age groups was 0.20 log units (photopic) and 0.33 log units (mesopic). Loss in contrast sensitivity due to luminance (two successive triplets) increased slightly with age. Both photopic and mesopic letter contrast sensitivity significantly improved as photopic visual acuity increased. CONCLUSIONS: Under mesopic conditions, Pelli Robson contrast sensitivity began to decline 1 decade earlier than under photopic conditions and was affected by visual acuity. Normal values for mesopic contrast sensitivity could be of help in deciding whether mesopic function is normal or a decrease in contrast sensitivity is pathologic in nature. PMID- 15523962 TI - Dapiprazole for night halos caused by angle-supported phakic intraocular lenses. PMID- 15523963 TI - Use of illuminance and luminance units. PMID- 15523964 TI - Single-value metrics of wavefront aberrationare we there yet? PMID- 15523965 TI - Improving visual function diagnostic metrics with the use of higher-order aberration information from the eye. AB - PURPOSE: This paper reviews the currently used visual function diagnostic metrics, acuity, refractive error, and contrast sensitivity, and suggests ways to create new metrics using the information that has recently become available due to advances in measuring the higher-order aberrations of the eye. Particularly, emphasis is placed on finding metrics that address certain aspects of vision rather than on general metrics. METHODS: Two metrics based on the modulation transfer function are introduced, the Visual Quality Factor (VQF), which is a value based on the modulation transfer function between the spatial frequencies of 3 and 12 cycles per degree (c/deg) giving a measure of the overall degradation of visual quality due to aberrations, and the Subjective Sharpness Factor (SSF), which is a value based on the modulation transfer function between the spatial frequencies of 15 and 40 c/deg, giving a measure of the decrease in perceived image "sharpness" due to aberrations. Two metrics based on the point spread function are suggested, Point Spread Quality (PSQ), which is a measure of the "compactness" of the point spread, and Multiplicity Factor, which is a measure of the multiplicity of perceived images by measuring the number of discrete peaks in the point spread function. RESULTS: The VQF and SSF values for both monochromatic and polychromatic cases are analyzed with regard to the number of acuity letters lost using data from a published study. CONCLUSIONS: The SSF is found to be the best overall correlate with acuity performance if the degradation of the letter stimuli is not excessive. PMID- 15523966 TI - The importance of the phase transfer function to visual function and visual quality metrics. AB - PURPOSE: To propose a vision quality metric that directly incorporates the phase transfer function. METHODS: The process of retinal image formation from the pupil to the retina is presented to identify possible domains for calculating vision quality metrics. These domains are pupil plane metrics using ocular aberrations, and retinal plane metrics using the point spread function or optical transfer function. The importance of the phase transfer function to visual performance is demonstrated and a method is proposed for directly including phase in a vision quality metric based on ocular wavefront error. RESULTS: A family of vision quality metrics is proposed, which directly incorporates the Stiles-Crawford effect, contrast sensitivity function, and the phase transfer function. CONCLUSION: The phase transfer function is an important aspect of the quality of an image, and it may be directly included in metrics intended to quantify vision quality. PMID- 15523967 TI - Measuring visual acuity--mesopic or photopic conditions, and high or low contrast letters? AB - PURPOSE: To develop single-valued wavefront aberration metrics that correlate strongly with visual performance. The purpose of this study is to explore whether photopic high contrast visual acuity (VA) is an appropriate visual performance reference and whether mesopic and/or low contrast testing provides any advantage. METHODS: Subjects from the Texas Investigation of Cataract Optics study (N = 148) ranged in age from 21.6 to 83.8 years and from clear lens to dense nuclear cataract. Visual acuity was measured under four conditions: photopic high (VA(PHC)) and low (VA(PLC)) contrast, mesopic high (VA(MHC)) and low (VA(MLC)) contrast (photopic = 280 cd/m(2), mesopic = 0.75 cd/m(2)). Variables were tested for compliance with normality (-2.00 < skew and kurtosis < 2.00) and transformed if required. Linear regression and Bland-Altman 95% limits of agreement (+/-1.96 SD) were used to examine relationships between VA variables and between VA and wavefront aberration metrics. RESULTS: The two photopic measures VA(PHC) (skew = 2.57, kurtosis = 12.2) and VA(PLC) (1.67, 5.41) were poorly distributed, but the two mesopic measures VA(MHC) (0.88, 1.67) and VA(MLC) (0.29, -0.18) were normally distributed. Strong correlations existed between the (transformed) VA variables (R: 0.53 to 0.84). However, limits of agreement ranged over 0.30 to 0.42 logMAR, whereas retest data suggested a range of 0.15. All four were offered in stepwise multiple linear regression for 30 wavefront metrics: 20 included VA(MLC) alone, two included VA(PLC), two included VA(MHC) and six included both VA(MLC) and VA(MHC); R(2) averaged 25%. CONCLUSION: Although strongly correlated, low contrast and/or mesopic VA testing provides different information. Wave aberration metrics correlates better with VA(MLC) making this the visual performance test of choice. PMID- 15523968 TI - Quantifying scatter in Shack-Hartmann images to evaluate nuclear cataract. AB - PURPOSE: Quantify and localize lenticular forward scatter using Shack-Hartmann wavefront sensing (SHWS) as single-valued metrics and a scatter map, and to examine the relationships between forward scatter and backscatter metrics and visual acuity. METHODS: We obtained SHWS images from 148 patients in the Texas Investigation of Cataract Optics study. Patient age was 22 to 84 years, with Lens Opacities Classification System III (LOCS III) nuclear opalescence (NO) scores ranging from 0.8 to 5.6. Visual acuities were measured at photopic (280 cd/m2) high (VA(PHC)) and low contrast (VA(PLC)) and mesopic (0.75 cd/m2) high (VA(MHC)) and low contrast (VA(MLC)). Scattering was described in a scatter map and by five single-valued metrics characterizing SHWS lenslet point spread functions. The relationships between scatter and visual acuity were tested using linear regression. RESULTS: Visual acuities decreased proportional to both LOCS III NO (R2=up to 39%) and scatter metrics (R2=up to 21%). Stepwise multiple linear regression improved visual acuity prediction by including a backscatter and a forward scatter metric (R2 up to 51.2%). For the subjects over age 60 years (N=46, 68.8+/-6.12 years), the forward scatter metrics explain as much variance in visual acuities (R2=up to 29%) as LOCS III NO (R2=up to 26%). Combined they accounted for up to 48.8% of visual acuity variance. CONCLUSION: Forward light scatter can be quantified using SHWS and the resulting metrics explain significant variance in visual acuity, especially in the aging eye. Together with a backscatter metric they explain approximately 50% of the variance in VA. PMID- 15523969 TI - Vision improvement by correcting higher-order aberrations with phase plates in normal eyes. AB - PURPOSE: To psychophysically demonstrate vision improvement when correcting higher-order aberrations with phase plates in normal eyes. METHODS: The wavefront aberrations of three nonsurgical normal eyes were measured with a Shack-Hartmann wavefront sensor. With these measured aberrations, phase plates were fabricated using a lathing technique. Theoretical improvement in retinal image quality was estimated by calculating the optical modulation transfer functions under the white light condition. Visual acuity measurements were also conducted to demonstrate improvement in visual performance after correcting higher-order aberrations with the phase plate. In this visual acuity measurement, a tumbling "E" with high (100%) and low (10%) contrast was used. RESULTS: The phase plate reduced the higher-order root mean square (RMS) wavefront error from 0.39 +/- 0.09 to 0.15 +/- 0.02 microm (mean +/- standard deviation from three eyes) for a 6-mm pupil. With the phase plate, retinal image quality based on the volume of modulation transfer function under 60 cycles per degree (c/deg) was improved by a factor of 1.8 +/- 0.4 over that of the eyes with spherocylindrical correction only. Average improvement in visual acuity achieved by correcting the higher order aberration was 0.23 lines with high-contrast letters and 1.12 lines with low-contrast letters. All subjects reported subjective improvement in image quality of the letter with the phase plate. CONCLUSION: The phase plate effectively corrected the higher-order aberrations in normal eyes. As a result, both retinal image quality and visual acuity especially with the low-contrast letters were improved. This study demonstrated the feasibility of correcting higher-order aberrations and improving vision with customized optics. PMID- 15523970 TI - Method of determining a patient's subjective refraction based on objective measurement. AB - PURPOSE: To develop a method for predicting the manifest refraction from objective measurement using wavefront sensing. We tested the hypothesis that, when the eye has an optimal refractive correction, the refractive power of the eye is in the most stable state, with the least fluctuation in its total optical aberration and hence least amount of standard deviation in the wavefront error. METHODS: Nine patients were enrolled in this study. All measurements were performed using a grating-based wavefront instrument manufactured by Ophthonix, Inc. The test procedure involved adding defocus power binocularly in the line of sight of the patient. Five added diopter powers were used: plano (no add), +1.00 D, +0.50 D, -0.50 D, and -1.00 D. The eyes were undilated, and the measurement was taken at a single added defocus power each day over 5 days. The patients had a binocular view and were asked to focus on a picture at 20 feet. Manifest refraction was used as the reference point (the +/- added powers are relative to this "plano" power). RESULTS: For each patient, the standard deviation (STDEV) of the total root mean square values of the wavefront measurement were used as a measure of vision fluctuation. Spherical equivalent manifest refraction was tested against the added defocus as a measure of the method's predictability. Graphic plots of standard deviation of the total wavefront error against the added diopter power showed that the minima of the vision fluctuation from the data sets were within -0.10 and +0.30 D (95% limit of agreement). The mean difference between the manifest refraction and the most stable point of vision was 0.14 D. CONCLUSIONS: The preliminary results of 10 eyes show promise that using an objective method of monitoring vision fluctuation may produce good agreement with the manifest refraction. The method should be further validated using a smaller step of added defocus and involving a larger number of test subjects representing a sample of the general population. PMID- 15523971 TI - Unresolved issues in the prediction of subjective refraction from wavefront aberration maps. AB - Sources of uncertainty in the conversion of wavefront aberration maps to refractive prescriptions are described. These uncertainties highlight the nature of optimum corrections, the imprecise far point in human eyes, multiple strategies for locating an approximate far point, and unresolved aspects of the gold standard for judging success. The conclusion points to future experiments needed to resolve these uncertainties to achieve more accurate and precise methods of objective wavefront refraction. PMID- 15523972 TI - Advantages and disadvantages of the Zernike expansion for representing wave aberration of the normal and aberrated eye. AB - PURPOSE: Zernike expansion has been selected for use in describing wavefront aberrations in the human eye. The advantages and limitations of this approach are assessed for eyes with varying degrees of aberration. METHODS: Corneal topography examinations were taken with the Nidek OPD-Scan topographer/aberrometer. These higher data density corneal topography examinations were converted to height data and subsequently to wavefront representations. System noise was evaluated with a 2D frequency analysis of 43-D test balls. Both Zernike polynomials and 2D Fourier transforms were used to evaluate fidelity in the presentation of the point spread function. A display format for potential clinical use was developed based upon Zernike decomposition. RESULTS: Systematic noise from the corneal topographer was found to be minimal and, when eliminated, produced small changes in the point spread function. Using Zernike decomposition up to the 30th order failed to preserve the higher frequency aberrations present in aberrated eyes. Use of a Zernike decomposition display with a fixed micron scale presented only clinically significant details of spherical aberration, coma, trefoil, irregular components above third order and total higher-order aberrations (above second order). CONCLUSIONS: Zernike polynomials excel in extracting the low-order optical characteristics of visual optics. Zernikes accurately represent both low- and high-order aberrations in normal eyes where high-order aberrations are clinically insignificant. For eyes after corneal surgery or eyes with corneal pathology such as keratoconus that have significant higher-order aberrations, the Zernike method fails to capture all clinically significant higher-order aberrations. PMID- 15523973 TI - Ablation design in relation to spatial frequency, depth-of-focus, and age. AB - PURPOSE: To consider factors relevant to the design of wavefront-aberration-based customized ablations. METHODS: Review. RESULTS: Ablations that seek to eliminate all wavefront aberrations, both second- and higher-order, may not be optimal for all patients. This is particularly the case for presbyopes. Their main requirement will normally be for extended binocular depth-of-focus to yield adequate distance and near vision with good retinal contrast at lower spatial frequencies, rather than the highest levels of acuity and modulation transfer function at a single distance. For many presbyopes, this can be achieved by aiming for monovision correction or low myopic astigmatism, with reasonable but not necessarily complete correction of higher-order aberrations. This compromise allows a range of everyday tasks to be carried out, including face recognition and reading. CONCLUSION: The targeted correction of wavefront aberration should take into account the visual needs and preferences of the individual patient. PMID- 15523974 TI - Transferring wavefront measurements into corneal ablations: an overview of related topics. AB - We give an overview of possible side effects that are specific for, or of particular relevance in, customized treatments. Certain processes involved in customized ablations have the potential to alter the quality of the optical correction. Professionals associated with customized treatment should be informed and trained with respect to possible sources of error. PMID- 15523975 TI - Custom phototherapeutic keratectomy with intraoperative topography. AB - PURPOSE: Treatment of highly aberrated eyes with opacities or irregularities consequent to previous refractive treatment, ulcers, keratitis, trauma, or corneal dystrophies remains a challenge for refractive surgeons. We evaluated the results of custom phototherapeutic keratectomy (CPK) with intraoperative corneal topography-based aberrometry and custom ablation in highly aberrated eyes. METHODS: We prospectively evaluated eyes undergoing custom phototherapeutic keratectomy (CPK) with custom ablation for corneal opacities and/or irregularities due to previous refractive treatment, keratitis, trauma, or dystrophies. Customization was planned according to corneal topography-based aberrometry, performed intraoperatively after removal of corneal epithelium. RESULTS: We treated 35 eyes of 34 patients. Mean preoperative haze was 1.6 +/- 1.4 in eyes with previous refractive surgery. Mean preoperative pachymetry was 425.7 +/- 119.3 microm. Mean preoperative spherical equivalent refraction was 1.60 +/- 3.65 D ranging from -9.38 to +10.00 D. No eye reached 20/15 best spectacle-corrected visual acuity (BSCVA), and only 63% could see 20/30 BSCVA. At final 6-month examination, mean spherical equivalent refraction was -0.71 +/- 4.01 D, ranging from -11.25 to +5.25 D. All eyes reached 20/30 or better BSCVA, and 19% had 20/15 or better BSCVA. CONCLUSIONS: Intraoperative, epithelium-free topography-based corneal aberrometry proved to be a feasible option for custom ablation in highly aberrated eyes. This approach widens the applications of PTK, providing a new procedure, custom PTK, that can be a successful solution for eyes that would otherwise require penetrating keratoplasty. PMID- 15523976 TI - Wavefront-guided laser epithelial keratomileusis with the wavelight concept system 500. AB - PURPOSE: Laser epithelial keratomileusis (LASEK) is a technique of refractive surgery for treatment of myopia. More than 90% of patients are within the intended correction of +/-0.50 diopters (D), without any unwanted side effects. After conventional refractive surgery, higher-order aberrations are induced. To reduce these aberrations, we used wavefront-guided ablation. METHODS: Twenty patients with visual problems in ambient light were included. The primary surgery was 141 to 463 days before the retreatment. LASEK was performed with the Camellin technique with 20% EtOH for 30 seconds. Before surgery, the root mean square (RMS) values were measured and the laser treatment was performed with the WaveLight Concept System 500 (WaveLight Laser Technologie AG). The preoperative spherical equivalent refraction was -1.22 D (SD 0.57 D) and the astigmatism was 0.42 D (SD 0.25 D). Preoperatively and 6 months postoperatively, a comprehensive examination was performed. RESULTS: All retreatments were performed without any intra- or postoperative complications. All patients reported an improvement of vision. The 6-month postoperative refraction was 0.25 D (SD 0.36 D). RMS decreased significantly from 0.72 to 0.47. RMS fourth order also decreased significantly from 0.60 to 0.30 and the spherical aberration decreased significantly from 0.57 to 0.28. CONCLUSIONS: In earlier studies, patients reported reduction of contrast vision, even after resurgery. Our patients showed a significant reduction of higher-order aberrations and better contrast vision after retreatment with wavefront guided LASEK. Retreatment with wavefront-guided LASEK has the potential to correct night vision problems after excimer laser surgery. PMID- 15523977 TI - Effect of higher-order wavefront aberrations on binocular summation. AB - PURPOSE: To determine the effect of individual Zernike wavefront aberrations on binocular summation and binocular visual acuity. METHODS: A 0.25-microm wavefront aberration of second, third and fourth order Zernike modes were introduced into a set of log minimum angle of resolution unit (logMAR) visual acuity charts convolved by CTView. Subjects were dilated and fitted with an artificial pupil of 3 mm. For each set of charts, right eye, left eye, and binocular acuity was measured. The gain in binocular visual acuity over monocular visual acuity was defined as binocular summation. The visual acuity lost binocularly reading aberrated charts was normalized for each subject and defined as the aberration induced loss in acuity. RESULTS: Binocular summation was 10.0% (95% limits of agreement 8.8 to 11.1%) in the unaberrated state and ranged from 17.3 to 3.4% in the Zernike modes studied. Binocular summation was greatest in defocus followed by coma and astigmatism. The aberration induced loss in monocular and binocular acuity was higher for Zernike modes with low angular frequency compared to those with high angular frequency. Linear regression showed a significant relationship between aberration-induced loss of visual acuity and binocular summation. CONCLUSIONS: A fixed amount of root mean square (RMS) aberration has a varied effect on binocular vision depending on the angular frequency and radial order of Zernike mode. Binocular vision has a positive effect in reducing the visual impact of aberrations as Zernike modes that suffer from the most loss of visual acuity also experience the greatest amounts of binocular summation. PMID- 15523978 TI - Corneal thickness and elevation maps computed from optical rotary scans. AB - PURPOSE: Recently, the authors presented a technique that allows corneal thickness measurements along any meridian from optical sections obtained using a rotary scanning system. This paper presents three-dimensional mapping of the corneal thickness and topography of both corneal surfaces, obtained with the rotary system. METHOD: Corneal thickness and topography are computed from optical sections obtained by illumination with a collimated beam expanded in a fan by a small cylindrical lens. This lens is provided with motor driven rotation to perform automated rotary scanning of the whole cornea. Two cameras are used to capture the images of the optical sections. RESULTS: With this system, it is possible to obtain measurements of corneal thickness, as well as corneal topography. Corneal thickness and elevation maps are shown. CONCLUSIONS: Although still under development, this new optical system allows measurement of the thickness of the whole cornea as well as topographical mapping of both corneal surfaces. PMID- 15523979 TI - Corneal asphericity and retinal image quality: a case study and simulations. AB - PURPOSE: The optical quality of retinal images is dependent on the refracting elements of the eye including the nominally aspheric cornea and crystalline lens. This paper presents a retrospective theoretical analysis of the impact of corneal asphericity on the quality of retinal images. Clinical data are from the VISX Incorporated CustomVue IDE. METHOD: Topography, contrast sensitivity, and visual acuity data were collected from 278 myopic eyes before and after wavefront-guided laser surgery. The measured corneal surface of each eye was fitted to a conic, and a Q-value was computed for a 5.5-mm pupil. A model eye was used to simulate various amounts of optical asphericity. RESULTS: Preoperatively, most corneas exhibited negative conic shape constants. Postoperatively, corneas were about equally divided between positive and negative conics. There was no statistically significant correlation between the shape of the cornea and the subjects' perceptions of image quality including contrast sensitivity and visual acuity. Simulations showed that the corneal Q-value can vary from more to less prolate depending upon the shape of the internal surface. CONCLUSION: Following wavefront guided laser in situ keratomileusis (LASIK), contrast sensitivity is usually good and is not dependent upon the corneal conic shape. Better visual outcomes are more likely with a customized shape than a standard best conic shape. PMID- 15523980 TI - Understanding pre-market approval and labeling differences of two leading customized ablation platforms: a call for reform at the FDA. AB - PURPOSE: To analyze the Food and Drug Administration (FDA) pre-market approval reported data for two currently available wavefront-guided excimer laser systems. METHODS: VISX and Alcon myopic wavefront laser in situ keratomileusis (LASIK) trials were analyzed based on public information available in the pre-market approval documents. RESULTS: Clinical tests and outcomes reporting varied between pre-market approvals. One pre-market approval reported contrast sensitivity whereas the other did not, although data was collected in both studies. Based on the FDA definition of "accountability," one pre-market approval reported 12-month accountability of 97.7% although only 24.5% of patients were analyzed at that interval. One pre-market approval reported 6-month results whereas the other reported 6- and 12-month results. One pre-market approval reported grouped pre- and postoperative subjective survey results; the other reported individual changes in pre- to postoperative subjective symptoms. CONCLUSION: We recommend an FDA revision to the definition of "accountability" and also recommend future FDA refractive surgery trials be guided by standardized criteria. PMID- 15523981 TI - The pupil is a moving target: centration, repeatability, and registration. AB - PURPOSE: To evaluate the ability to accurately center and register wavefront data to provide effective custom wavefront application for vision correction. METHODS: Centration images were obtained of 40 undilated pupils at 5 different illumination levels using the Alcon LADARWave aberrometer (Alcon Laboratories Inc, Fort Worth, Tex). Pupil position relative to the limbus was determined for all eyes under all levels of illumination. Variability in centration of the pupil was based on difference in magnitude of offset between pupil center and limbal center. Human operators were compared to a computerized limbus recognition system through repeatability testing using 10 different images of one eye of a single patient. RESULTS: As determined by human operators, the pupil center in undilated pupils shifted a mean of 175 microm between the lowest level and the highest level of illumination (range 34 to 335 microm [standard deviation 84 microm]). Repeatability testing suggests human measurements can accurately position the pupil to within approximately 50 microm using current technology. Computer image processing using limbus recognition software appears equal or superior to manual pupil centration. CONCLUSION: The pupil center can have a significant change in position with changing illumination. Wavefront data must be centered on a fixed eye structure rather than the pupil center to ensure that wavefront data captured in scotopic light are appropriately matched to the pupil in the photopic light used at surgery. The goal is to avoid clinically significant decentration of the wavefront between capture and excimer laser application. PMID- 15523982 TI - The theoretical effect of measured wavefront diameter on estimating peripheral wavefront data. AB - PURPOSE: To compare the difference between wavefront data measured in the periphery and that calculated from estimations using a smaller measured region. METHODS: A total of 32 eyes were measured using the Alcon LADARWave aberrometer (Alcon Laboratories Inc, Fort Worth, Tex). Aberrations were measured through a dilated pupil to the 6th order at a 6.5-mm diameter. Data were then "clipped" to 5.0, 5.5, and 6.0 mm, and each of these smaller diameter wavefronts was used to estimate the larger diameter wavefront coefficients. The measure of interest was the mean and range of error between the estimated and measured values, and how they varied for the different diameters of known data. RESULTS: The differences in both the mean and range of error were inversely correlated to the diameter of the region of known data. As this region grew smaller, the region estimated grew larger, with resultant larger mean errors. At 5 mm, the estimated zone was an annulus of 0.75-mm width; in this case the estimated higher order aberrations were 60% higher than those measured, with a range of 40% lower to 300% higher. At 5.5 mm and 6.0 mm, these differences were much smaller, but statistically significant. The differences were greatest when significant peripheral aberrations were measured, as can occur with post-laser in situ keratomileusis eyes. CONCLUSION: Basing a treatment on an estimation of the peripheral wavefront may lead to an inaccurate ablation profile. The inaccuracy increases as the size of the measured wavefront decreases relative to the planned ablation zone. The inaccuracy is also greatest when significant peripheral aberrations are present. Refractive surgery systems should only treat optical zones that are smaller than or equal to the diameter of the measured wavefront. PMID- 15523983 TI - Contralateral comparison of Alcon CustomCornea and VISX CustomVue wavefront guided laser in situ keratomileusis: one-month results. AB - PURPOSE: To evaluate the differences in outcomes between CustomCornea and CustomVue LASIK treatments on different eyes of the same patient 1 month after surgery. METHODS: Thirty-five patients were enrolled in two phases of a prospective study and treated with Alcon CustomCornea LASIK (Alcon Laboratories Inc, Fort Worth, Tex) on one eye and VISX CustomVue LASIK (VISX, Santa Clara, Calif) on the fellow eye. Twenty-five patients were treated in phase I without nomogram adjustments and ten patients were treated in phase II with nomogram adjustments made to the VISX CustomVue treatments only. Standard clinical outcomes such as visual acuity and manifest refraction were evaluated as well as quality of vision measures such as subjective questionnaires, contrast sensitivity, and higher order optical aberrations. RESULTS: In the phase I group, 92% of the CustomCornea eyes and 72% of the CustomVue eyes were within 0.50 diopters (D) of their attempted correction. In the phase II group, with a nomogram applied to the CustomVue eyes, 80% of the CustomCornea eyes and 100% of the CustomVue eyes were within 0.50 D of their attempted correction. There was no loss of 22 lines of best spectacle-corrected acuity for the CustomCornea or CustomVue eyes in either phase I or phase II. Wavefront analysis demonstrated that the CustomVue eyes had statistically significantly more spherical aberrations at 1 month compared to the CustomCornea eyes in the phase I and II groups. An average increase in mesopic contrast sensitivity at spatial frequencies was seen for the CustomCornea eyes in phase II and in only one spatial frequency for the CustomVue eyes in phase II. CONCLUSION: At 1 month after LASIK, the CustomCornea eyes showed superiority in the number of eyes seeing 20/20 or better uncorrected visual acuity, constrast sensitivity at 3 and 12 cpd, and the amount of postoperative spherical aberration. After nomogram adjustment, CustomVue eyes had a more accurate refractive outcome. Patients were satisfied or extremely satisfied with both systems, 92%-100%. PMID- 15523984 TI - Wavefront-guided laser in situ keratomileusis with the Alcon CustomCornea and the VISX CustomVue: three-month results. AB - PURPOSE: To evaluate and compare the visual and clinical outcomes of wavefront guided laser in situ keratomileusis (LASIK) with the Alcon CustomCornea (Alcon Laboratories Inc, Fort Worth, Tex) and VISX CustomVue (VISX, Santa Clara, Calif) systems. METHODS: Ninety-three eyes of 56 patients (50 and 43 consecutive eyes on CustomCornea and CustomVue, respectively) were enrolled in a prospective multisurgeon clinical outcome study. Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), and manifest refraction were measured at 1 day, 1 week, 1 month, and 3 months postoperatively. Early treatment diabetic retinopathy study (ETDRS) visual acuity and contrast sensitivity were measured at 1 month and 3 months, and wavefront mapping at 3 months. RESULTS: Preoperatively, the CustomCornea group had a mean sphere of -3.90 +/- 1.62 diopters (D) (range 0.50 to -7.25 D), mean cylinder of +0.62 +/- 0.39 D (range 0 to +1.50 D), and mean manifest spherical equivalent refraction of -3.59 +/- 1.54 D. The CustomVue group had a sphere of -3.87 +/- 1.45 D (range -1.75 to -6.75 D), cylinder of +0.49 +/- 0.36 D (range 0 to +1.50 D), and manifest spherical equivalent refraction of -3.62 +/- 1.46 D. At 3 months, 98% of the CustomCornea group and 95% of the CustomVue group were within +/-0.50 D. Ninety-nine percent of eyes did not change >0.50 D (manifest spherical equivalent refraction) between 1 month and 3 months. CustomCornea eyes improved on contrast sensitivity testing and had a better profile than CustomVue for 20/15 Snellen and 20/12.5 ETDRS acuity. Both laser groups had a decrease in higher order aberrations with statistical significance for coma and spherical aberration in the CustomCornea group. CONCLUSION: Wavefront-guided LASIK with both systems is safe and effective. PMID- 15523985 TI - Randomized comparison of custom laser in situ keratomileusis with the Alcon CustomCornea and the Bausch & Lomb Zyoptix systems: one-month results. AB - PURPOSE: To compare the early visual results of custom laser in situ keratomileusis (LASIK) outcomes for two different excimer laser systems. METHODS: This is a preliminary report of an ongoing prospective study that evaluated 30 patients (60 eyes) who were randomized to one of two custom LASIK treatment groups: 15 patients (30 eyes) with the Alcon CustomCornea laser system (Alcon Laboratories Inc, Fort Worth, Tex) and 15 patients (30 eyes) with the Bausch & Lomb Zyoptix laser system (Bausch & Lomb, Rochester, NY). These matched patients were required to qualify for treatment with both custom laser systems. All flaps were created with the IntraLase femtosecond laser (IntraLase Corp, Irvine, Calif) using a superior hinge. Early outcome measures are uncorrected visual acuity (UCVA) at 1 day, 1 week, and 1 month postoperative; manifest refractions at 1 month follow-up; and a comparison of 1-month postoperative UCVA to preoperative best spectacle-corrected visual acuity (BSCVA). Wavefront, topography, contrast sensitivity, and subjective questionnaires were also performed and will be reported after 3-month follow-up. RESULTS: At 1 month, the percentage of eyes with UCVA of 20/20 and 20/40 were similar with 93% and 100%, respectively, for CustomCornea eyes and 90% and 97%, respectively, for Zyoptix eyes. A difference between the two groups is in the percentage of eyes with UCVA of 20/16, 20/12.5, and 20/10 with 80%, 47%, and 13%, respectively, for CustomCornea and 70%, 10%, and 0%, respectively, for Zyoptix. The mean residual spherical equivalent refraction was +0.01 +/- 0.34 diopters (D) for CustomCornea and -0.04 +/- 0.38 D for Zyoptix. The two groups are similar at +/-1.00 D of intended correction, with CustomCornea 100% and Zyoptix 97%. However, +/-0.25 D the CustomCornea group was 10% more accurate at 73% versus 63% for the Zyoptix group. The efficiency ratio (postoperative UCVA compared to preoperative BSCVA) was greater for the CustomCornea group at 20/20, 20/16,20/12.5, and 20/10 levels. CONCLUSION: Custom LASIK with CustomCornea and Zyoptix is safe and effective and produces excellent early visual outcomes. This study demonstrates that current standard metrics for reporting refractive surgery visual outcomes are not adequate for evaluating wavefront-guided laser vision correction procedures. Amending the standard reporting metrics to include visual outcomes at the 20/16, 20/12.5, and 20/10 levels; accuracy at the +/-0.25 D of attempted; and postoperative UCVA compared to preoperative BSCVA will improve our ability to analyze wavefront-guided outcomes. PMID- 15523986 TI - Expanded range customcornea algorithms for myopia and astigmatism: one-month results. AB - PURPOSE: To evaluate the early clinical results achieved with an algorithm adjusted for an expanded range of correction in wavefront-guided customized ablation with the LADARVision4000 (Alcon Laboratories Inc, Fort Worth, Tex). METHODS: Fifty-five consecutive eyes from 31 patients underwent wavefront-guided, customablation laser in situ keratomileusis (LASIK) (Hansatome and BD 4000 microkeratomes, LADARWave aberrometer, LADARVision4000 laser system). These were normal myopic and astigmatic eyes that had never been operated on. The spherical equivalent refractive error was +0.30 to -8.13 diopters (D) (mean -4.26 +/- 2.14 D) and the astigmatism ranged from 0 to -3.75 D (mean -0.97 +/- 0.96 D). eyes were treated according to the CustomCornea protocol using the commercial LADARVision4000 platform. Patients were followed for at least 2 months and standard visual measurements were taken and recorded at 1 day and 1, 3, and 6 months after treatment. The results of treatment were assessed using the following parameters: uncorrected visual acuity (UCVA) and best spectacle corrected visual acuity (BSCVA); manifest spherical equivalent refractive error; wavefront measurement of high order aberrations; and subjective reports of visual symptoms by patients. RESULTS: One month after LASIK, the mean manifest spherical equivalent refractive error was -0.18 +/- 0.41 D. Seventy-eight percent and 96% of eyes had manifest spherical equivalent refractive error within +/-0.50 D and +/-1.00 D of attempted correction, respectively, with 78% of eyes with 20/20 or better UCVA. Eyes with 20/16 and 20/12.5 BSCVA were 89% and 36%, respectively (55% and 7% preoperatively). The mean high order aberrations root-mean-square was 0.34 +/- 0.10 microm preoperatively and 0.35 +/- 0.09 microm postoperatively (6.0 mm pupil size). High order aberrations were either reduced, unchanged, or increased by <10% in 58% of eyes. Subjectively, patients reported no visual symptoms and had no complaints regarding the quality of their vision. CONCLUSION: The new optimized algorithm for higher refractive errors appears to be effective in improving BSCVA, and a minority of patients demonstrated an increase in the magnitude of high order aberrations when compared to preoperative aberrometry. A slight spherical equivalent refraction undercorrection will be addressed with future nomogram adjustments. PMID- 15523987 TI - Alcon CustomCornea wavefront-guided retreatments after laser in situ keratomileusis. AB - PURPOSE: To evaluate the outcome of wavefront-guided ablations for the correction of residual myopia and astigmatism after standard laser in situ keratomileusis (LASIK). METHODS: Twenty nine eyes of 26 patients who underwent wavefront-guided LASIK retreatment with Alcon CustomCornea (Alcon Laboratories Inc, Fort Worth, Tex) were evaluated. Complete ophthalmologic examination, corneal topography, and wavefront measurements were performed. Uncorrected visual acuity (UCVA), best spectacle-corrected visual acuity (BSCVA), manifest refraction, and wavefront analysis were evaluated preoperatively, 1 week, 3 months, and 6 months after surgery. Wavefront measurements were assessed using the LADARWave device. Statistical analysis was performed using the McNemar test and percentages of success. RESULTS: One week postoperatively, UCVA was > or =20/40 in 100% of eyes (> or =20/20 in 31%) and BSCVA was > or =20/40 in 100% (> or =20/20 in 73%). Wavefront analysis showed a decrease in total aberrations, high order aberrations, defocus, coma, spherical aberration, and other terms of higher order aberrations at 1-week follow-up. Three months postoperatively, UCVA was > or =20/40 in 100% of eyes (> or =20/20 in 38%) and BSCVA was > or =20/40 in 100% (> or =20/20 in 81%). Six months postoperatively, UCVA was > or =20/40 in 100% of eyes (> or =20/20 in 60%) and BSCVA was > or =20/40 in 100% (> or =20/20 in 90%). Wavefront analysis showed decrease in total aberration, high order aberration, defocus, coma, and spherical aberration. CONCLUSION: Wavefront-guided LASIK retreatment in post-LASIK eyes represents a good option for laser vision correction. All eyes showed reduction in pre-existing total aberrations. Some high order aberration components decreased in this initial series. Further follow up is necessary to assess the initial predictability of wavefront-guided LASIK upgrade. PMID- 15523988 TI - Voluntary sector turns up the volume on disability services. PMID- 15523989 TI - Patient consent ideas. Two sides of the fence. PMID- 15523990 TI - Data briefing. Cardiac surgery. A new report allows comparison of surgeons' perforance but is cautious about how it is presented. PMID- 15523991 TI - Clinical management. Brittle battle. AB - Women aged 75 and over with fractures will get bone-saving treatment and drugs straight away. Women aged 65-75 with fractures will need a DEXA scan before treatment can start. Specialists are concerned that women under 75 will lose out because scanning is patchy around the UK. PMID- 15523992 TI - Working lives. Meet the new model army. PMID- 15523994 TI - HJS people: regional focus on Wales. Valley of the holes. PMID- 15523993 TI - Mental health. Electric avenues. PMID- 15523995 TI - Healthcare-acquired infections. The pound sterling 1bn bug. PMID- 15523996 TI - Healthcare-acquired infections. Clean sweep. PMID- 15523997 TI - Acoustic detection and communication by decapod crustaceans. AB - This paper reviews behavioral, physiological, anatomical, and ecological aspects of sound and vibration detection by decapod crustaceans. Our intent is to demonstrate that despite very limited work in this area in the past 20 years, evidence suggests that at least some decapod crustaceans are able to detect and use sounds in ways that parallel detection and processing mechanisms in aquatic and terrestrial vertebrates. Some aquatic decapod crustaceans produce sounds, and many are able to detect substrate vibration at sensitivities sufficient to tell of the proximity of mates, competitors, or predators. Some semi-terrestrial crabs produce and use sounds for communication. These species detect acoustic stimuli as either air- or substrate-borne energies, socially interact in acoustic "choruses," and probably use "calls" to attract mates. PMID- 15523998 TI - Agonistic behavior in naive juvenile lobsters depleted of serotonin 5,7 dihydroxytryptamine. AB - We have been exploring the role of serotonin in fighting behavior in lobsters using a specific model of agonistic behavior, the establishment of hierarchical relationships between pairs of socially naive juvenile lobsters. We selected this model because the behavior is easily evoked, readily quantifiable, and the effects of experience are eleminated by using socially naive animals. In these studies we injected a specific neurotoxin, 5,7-dihydroxytryptamine, into juvenile lobsters over a 4-week period and then measured the effects on fighting behavior. This treatment reduces the levels of serotonin in the nervous system and immunocytochemical studies show a dramatic reduction in neuropil staining for the amine. Control animals received vehicle injection alone. All injected animals were paired against larger or smaller non-injected opponents, and three successive 30-min fights were carried out and statistically analyzed. The results were surprising: As with elevations of serotonin, reduced levels of serotonin increased the amount of time animals engaged in fighting behavior. No significant effects were seen on who initiated encounters, who retreated first, or who the eventual winner would be. Thus, in this model, elevation or reduction of serotonergic function increases the tendency of animals to engage in agonistic encounters. PMID- 15523999 TI - Factors influencing the susceptibility of anurans to motion sickness. AB - We examined the propensity for motion sickness in five anuran species, concentrating our efforts on the treefrog Rhacophorus schlegelii, because it had shown the greatest susceptibility to motion sickness in a previous study. We used parabolic flight as our provocative stimulus and fed all specimens a known volume of food 1.5-3 h before flight. The presence of vomitus in a frog's cage was our indicator of motion sickness. Significantly more emesis was observed in flight exposed than in control R. schlegelii (P < 0.05). There was no sex difference in susceptibility to motion sickness (P > 0.5). Individuals that vomited were significantly larger (P < 0.02) than those that did not. Among microgravity treated frogs, those that vomited spent on average 85% more time airborne and tumbling in microgravity than those that did not vomit (P=0.031). Our data support the view that postural instability and sensory conflict are elements of motion sickness in anurans. Specifically, conflicts between tactile, vestibular and visual input seem essential for producing motion-induced emesis in anurans. Since the factors that induce motion sickness in R. schlegelii are the same ones that produce motion sickness in humans, arboreal frogs may be useful alternative models to mammals in motion sickness research. PMID- 15524000 TI - Activity of descending contralateral movement detector neurons and collision avoidance behaviour in response to head-on visual stimuli in locusts. AB - We recorded the activity of the right and left descending contralateral movement detectors responding to 10-cm (small) or 20-cm (large) computer-generated spheres approaching along different trajectories in the locust's frontal field of view. In separate experiments we examined the steering responses of tethered flying locusts to identical stimuli. The descending contralateral movement detectors were more sensitive to variations in target trajectory in the horizontal plane than in the vertical plane. Descending contralateral movement detector activity was related to target trajectory and to target size and was most sensitive to small objects converging on a direct collision course from above and to one side. Small objects failed to induce collision avoidance manoeuvres whereas large objects produced reliable collision avoidance responses. Large targets approaching along a converging trajectory produced steering responses that were either away from or toward the side of approach of the object, whereas targets approaching along trajectories that were offset from the locust's mid longitudinal body axis primarily evoked responses away from the target. We detected no differences in the discharge properties of the descending contralateral movement detector pair that could account for the different collision avoidance behaviours evoked by varying the target size and trajectories. We suggest that descending contralateral movement detector properties are better suited to predator evasion than collision avoidance. PMID- 15524001 TI - The antennal motor system of the stick insect Carausius morosus: anatomy and antennal movement pattern during walking. AB - The stick insect Carausius morosus continuously moves its antennae during locomotion. Active antennal movements may reflect employment of antennae as tactile probes. Therefore, this study treats two basic aspects of the antennal motor system: First, the anatomy of antennal joints, muscles, nerves and motoneurons is described and discussed in comparison with other species. Second, the typical movement pattern of the antennae is analysed, and its spatio-temporal coordination with leg movements described. Each antenna is moved by two single axis hinge joints. The proximal head-scape joint is controlled by two levator muscles and a three-partite depressor muscle. The distal scape-pedicel joint is controlled by an antagonistic abductor/ adductor pair. Three nerves innervate the antennal musculature, containing axons of 14-17 motoneurons, including one common inhibitor. During walking, the pattern of antennal movement is rhythmic and spatiotemporally coupled with leg movements. The antennal abduction/adduction cycle leads the protraction/retraction cycle of the ipsilateral front leg with a stable phase shift. During one abduction/adduction cycle there are typically two levation/depression cycles, however, with less strict temporal coupling than the horizontal component. Predictions of antennal contacts with square obstacles to occur before leg contacts match behavioural performance, indicating a potential role of active antennal movements in obstacle detection. PMID- 15524002 TI - Presynaptic effects of octopamine, serotonin, and cocktails of the two modulators on neuromuscular transmission in crustaceans. AB - The effect of the biogenic amines octopamine and serotonin, and of both amines combined (cocktails) on transmitter release at neuromuscular junctions of two crustaceans was studied. octopamine (10(-8) mol l(-1) to 10(-6) mol l(-1)) either enhanced or decreased evoked transmitter release through presynaptic effects. The results were identical for the slow and the fast excitor in the closer muscle of the crab, and for the excitor in the opener muscle of the crayfish. Application of serotonin always resulted in a strong increase of release. However, this potentiating effect of serotonin was reduced in strength by subsequent application of cocktails consisting of serotonin and octopamine. In all experiments, a cocktail of serotonin and octopamine was less effective than serotonin alone. The decrease in the mean quantal content m by octopamine was due to a reduction of the probability of release p. Since both amines are synthesized in the central nervous system and are released from neurohaemal organs into the haemolymph bathing the neuromuscular junctions, the results suggest that the two amines, when present together, modulate transmitter release in an antagonistic way, and that the level of the two determines synaptic efficacy. PMID- 15524003 TI - Pharmacological brain stimulation releases elaborate stridulatory behaviour in gomphocerine grasshoppers--conclusions for the organization of the central nervous control. AB - Grasshoppers produce a variety of sounds generated by complex movements of the hindlegs. Stridulation, performed in the context of partner finding, mating and rivalry, can be released by pressure injection of cholinergic agonists into the protocerebrum. Particularly stimulation with muscarinic agonists induced long lasting stridulation that resembled the natural behaviour to an astonishing degree, not only with respect to their temporal structure and right/left coordination, but also to changes in the song sequences according to the progress of courtship stridulation, even including accessory movements of other parts of the body. According to the complexity of their stridulatory behaviour ten gomphocerine species were chosen for this comparative study. The results indicate that the protocerebrum fulfils two important tasks in the control of stridulation: (1) it integrates sensory input relevant to stridulation that represents a certain behavioural situation and internal state of arousal, and (2) it selectively activates and deactivates the thoracic networks that generate the appropriate movement and sound patterns. With the knowledge of the natural behaviour and the accessibility to pharmacological and electrophysiological studies, the cephalic control system for stridulation in grasshoppers appears to be a suitable model for how the brain selects and controls appropriate behaviours for a given situation. PMID- 15524004 TI - [Introduction to asthma and chronic obstructive pulmonary disease pharmacoeconomics]. AB - It is important for the clinician to be familiar with the different aspects of cost in asthma and chronic obstructive pulmonary disease (COPD) care: direct, indirect, and intangible. In pharmacoeconomics the most common used analysis is cost-effectiveness measurements, where costs are compared to specific outcomes in both diseases. As health care costs continue to rise in asthma and COPD as well, it should be stressed that asthma therapy has changed from inpatient to ambulatory care, while the treatment of COPD to a higher degree still is based on inpatient care. PMID- 15524005 TI - [Epidemiology of bronchial asthma]. AB - Epidemiology of bronchial asthma is analyzed in many trials over the world. The prevalence of disease is dependent to genetic and environmental factors. Some connections have been shown in the paper. The data from international programs focused on allergic diseases have been presented in relation to our observations. PMID- 15524006 TI - [Epidemiology of chronic obstructive pulmonary disease]. AB - Chronic obstructive pulmonary disease (COPD) is a major cause of death and disability in many countries over the world. Prevalence of COPD is difficult to evaluate because of clinical similarity to bronchial asthma, and changes in diagnostic standards. Actually about 8-35% patients are suffering because of COPD. In Poland this problem was described only in some regional trials. For this reason the program for prevalence of COPD has to be established according to contemporary diagnostic and therapeutic standards. PMID- 15524007 TI - [Pharmacoeconomics in contemporary medicine]. AB - Following international examples, economic evaluation of health care programmes is starting to play an increasing role in Poland. Applying the broadest view, an economic evaluation is defined as the comparative analysis of alternative courses of action in terms of their costs and consequences. This definition includes economic evaluation of prescribed drugs by the term pharmacoeconomics. Pharmacoeconomics can be defined by the study of economic implications of drug usage. From a practical point of view, pharmacoeconomics is a part of health economics, the aim of which is to optimize pharmacotherapy through rational drug management. The role of pharmacoeconomic research is to identify measure and compare the resources consumed and corresponding costs with consequences in the form of clinical, humanistic and economic outcomes of pharmaceutical products and services used. The results of these studies serve as scientific evidence for decision making. The Polish Chapter of the International Society for Pharmacoeconomics and Research (ISPOR) is the main scientific organization operating in the field of pharmacoeconomics in Poland. PMID- 15524008 TI - [Clinical effectiveness of synergic therapy]. AB - The coadministration of long-acting inhaled beta2-agonists and inhaled corticosteroids is the most effective treatment for persistent asthma. It is often assumed that there is synergy between these medications. Many studied mostly extensively as a maintenance therapy for patients with persistent asthma, the combination inhaler is at least equivalent to its components administered separately and is superior to monotherapy with salmeterol/formoterol or inhaled corticosteroid in both pediatric and adult populations. The combination has a logical role in the treatment of moderate-to-severe asthma, offering the advantage of increased convenience and possibly improved compliance. In addition to improvements in lung function, symptom scores and quality of life, the combination therapy reduces exacerbation rates, an outcome that contributes to favorable cost-effectiveness. PMID- 15524009 TI - [Current recommendation for asthma treatment]. AB - Asthma is a chronic inflammatory disease, characterized chronic inflammation of respiratory tract, reversible bronchial obturation and non-specific bronchial hyperresponsiveness. Typical symptoms of asthma are cough, wheezing and dyspnea. Diagnosis should be based on positive reversibility test with beta2-agonist. In the management of asthma patient education, allergen avoidance and anti inflammatory treatment should be always implemented. Recently fast and long acting beta2-agonist was introduced with great success. Combination treatment with long acting beta2-agonists and topical glucocorticosteroids for patients with moderate and severe chronic asthma is recommended. We can also consider antileukotrienes and long acting theophylline as an option in patients not fully controlled by inhaled steroids. Patient with asthma should be always examined closely for chronic rhinitis and treated accordingly with antihistaminic drugs. Atopic asthma can be also treated with immunotherapy. With new drugs for asthma we can achieve full control of disease symptoms without unwanted side effects. PMID- 15524010 TI - [Treatment of bronchial asthma exacerbation]. AB - The purpose of this study was to present the actual possibilities of bronchial asthma exacerbation control. The role of viral infection, especially, human metapneumovirus infection may be a leading cause of disease exacerbation in children population. Prevalence of gastroesophageal reflux had been detected in 24% of asthmatic patients, in some cases as a cause of asthma exacerbation. Glucocorticosteroid therapy is accepted as cardinal in asthma exacerbation, but some adverse effects create a control limit. PMID- 15524011 TI - [Management of severe and difficult asthma]. AB - In about 5% of asthmatics patients despite of recommended by GINA 2002 treatment present sudden unexpected severe exacerbations. They are included into group of severe and difficult asthma. Based on the statement of Polish experts the diagnostic procedures and management of severe and difficult asthma was presented. PMID- 15524012 TI - [Asthma in children--epidemiology and management]. AB - The role of epidemiological surveillance is essential in early detection of asthma and prevention programs. The prevalence of asthma diagnosis increased significantly in children in last 20 years--conclusion and results of International Study of Asthma and Allergies in Childhood (ISAAC). The incidence of asthma in Polish children is almost 9%. The management in asthma in children is according to GINA 2000 and 2002 report. PMID- 15524013 TI - [Immunotherapy as the prevention of asthma and allergy]. AB - Paper describes anti-inflammatory and preventive action of immunotherapy in asthma and allergy. PMID- 15524014 TI - [Education of asthmatic patients]. AB - Asthmatics education is the essential part of their therapy--category A of clinical evidences. Authors show the advantages from the correctly conducted educational process and the world experiences in realization of preventive programmes. Possibilities of such procedure in Poland are underlined. PMID- 15524015 TI - [Costs of occupational asthma and chronic obstructive pulmonary disease]. AB - Many published data indicate that a relevant proportion of cases of asthma and chronic obstructive pulmonary disease (COPD) are caused by occupational exposure. That phenomenon results in many negative economic consequences, including, apart from considerable medical costs, also costs owing to unemployment and claiming for compensation, which are frequent in patients suffering from that illnesses. Hence, it is probable that increasing prevalence of those diseases may result in higher economical costs. The effectiveness of primary and secondary prevention in reduction of costs concerning occupational asthma and COPD has been proved. Only effective prophylaxis may decrease individual and public costs related to both diseases. PMID- 15524016 TI - [Pharmacoeconomics of asthma]. AB - Pharmacoeconomics is important in asthma as this condition continues to increase in prevalence throughout the world, consuming a larger and larger share of total health care budgets. Understanding how pharmacoeconomics can be applied in evaluating the cost-effectiveness of asthma treatments can be important in determining proper asthma care in a climate of limited resources in Poland. PMID- 15524017 TI - [Chronic obstructive pulmonary disease--the range of the problem, clinical stages]. AB - The prevalence of chronic obstructive pulmonary disease (COPD) increases worldwide, as well as in Poland. About 10% of adult population is suffering from COPD. COPD occurs mainly in smokers who are older than forty years. Diagnosis of COPD in case of many patients is not properly established. It refers mainly the patients in a mild or a moderate clinical stage. One-fourth of patients with COPD is suffering from a severe or an extremely severe clinical stage of COPD. When untreated, COPD is a fast progressing disorder, leading to respiratory disability and premature death. Therapy of COPD is related to the clinical stage of disease. PMID- 15524018 TI - [Complications of chronic obstructive pulmonary disease]. AB - The paper presents pathomechanisms and treatment of the complications of chronic obstructive pulmonary disease (COPD): ventilatory failure, infections, cor pulmonale, overlap syndrome and discusses acute life-threatening conditions. PMID- 15524020 TI - [Principles of chronic obstructive pulmonary disease treatment and patients' expectations]. AB - The article describes current, consistent with valid standards aims of treatment chronic obstructive pulmonary disease (COPD), focusing on perception of disease by patients. The methods of clinical status and patients' quality of life measurements have been introduced. PMID- 15524019 TI - [Perspectives in the management of chronic obstructive pulmonary disease]. AB - Results of clinical studies conducted in patients with chronic obstructive pulmonary disease (COPD) in recent years have pointed out at new perspectives in the management of COPD. They include both pharmacological and non-pharmacological methods. Progress in pharmacological therapy results mainly due to introduction of long acting anticholinergic compound and combined therapy with long acting beta2-agonists and inhaled corticosteroids. Non-invasive positive pressure ventilation (NIPPV) appears to play a significant role in the treatment of severe COPD exacerbations. Also pulmonary rehabilitation has found very important position in the management of COPD patients. However, from patients' perspective the most important benefit resulting from new therapies is an improvement in health related quality of life. PMID- 15524021 TI - [Causal proceeding in chronic obstructive pulmonary disease]. AB - At present, chronic obstructive pulmonary disease (COPD) is considered as serious clinical, social and economic problem. Widespread habit of smoking together with air-pollutions is the basic etiologic factors, whereas inflammatory process, which involves bronchial tree and pulmonary tissues, is the core of COPD pathogenesis. Quality of life has much been improved by the introduction of pharmacological treatment including: long-acting beta2-mimetics, cholinolytics, mucolytics and in severe cases inhaled glucocorticosteroids. Prophylactic immunization against influenza and pneumococci also has been proved to be helpful. Continuous patient education together with efforts promoting smoking cessation constitute the mainstay of non-pharmacological management. Chronic oxygen therapy, respiratory rehabilitation and non-invasive ventilation proved to be a practical alternative in patients with chronic respiratory failure. Lung volume-reduction surgery may sometimes be considered as an alternative to lung transplantation in very severe cases of COPD. PMID- 15524022 TI - [Exacerbations of chronic obstructive pulmonary disease]. AB - The problems of exacerbations of chronic obstructive pulmonary disease were presented in this study. The methods of diagnosis and treatment was discussed in this article. PMID- 15524023 TI - [Tiotropium as a controller of bronchoconstriction]. AB - Cholinergic nerve fibres arise in the nucleus ambiguus and the dorsal motor nucleus of the vagus nerve in the brainstem. They travel down as the vagus nerve to parasympathetic ganglia placed in the walls of the airways. From these ganglia, short postganglionic fibres innervate airway smooth muscle and the submucosal glands in the lung. Activation of vagal nerve releases acetylcholine at the neuroeffector junctions, where it binds to postsynaptic receptors, resulting in bronchoconstrictions. The resting bronchomotor tone in normal airways has a cholinergic component mediated via muscarinic cholinergic receptors. The human airways have five subtypes of muscarinic cholinergic receptors: the M1 and M3 mediate bronchoconstriction and stimulation of mucus secretion, while M2 control the release of acetylcholine from M1 and M3 receptors through a negative-feedback mechanism. Anticholinergic bronchodilators act by blocking muscarinic receptors. Tiotropium bromide is cutting age anticholinergic bronchodilator. It dissociates more slowly from M1 and M3 than from M2 receptors and subsequently has a long and safety duration of action. In COPD patients tiotropium comparing to placebo, ipratropium and long acting beta agonists significantly improves lung function. It is an effective bronchodilator that reduces dyspnea, COPD exacerbations frequency and improves health status. This suggests that tiotropium will make an important contribution to chronic pulmonary disease therapy. PMID- 15524024 TI - [Methods and cost effectiveness of nicotine abstinence]. AB - Tobacco smoking is responsible for many chronic diseases causing an important burden of morbidity, mortality and economic costs. The treatment of nicotine dependent smokers bases on nicotine replacement therapy and on use of bupropion. Rationales for treatment, methods and results of therapy as well as the cost effectiveness of these methods are described. PMID- 15524025 TI - [Pharmacoeconomics of vaccinations in chronic obstructive pulmonary disease]. AB - In this study basic types of pharmacoeconomics analysis are presented. Selected publications concerned influenza vaccinations in risk groups have been discussed. In age groups higher then 65 years costs of treatment are connected with rates of complications and hospitalisations. In COPD patients' pharmacoeconomic analysis shows positive results in more advanced age-groups. In Str. pneumoniae vaccination 23-valent vaccines are more clinically effective. COPD patients Streptococcus pneumoniae vaccination give better pharmacoeconomic results in higher age-groups. PMID- 15524026 TI - [Costs of chronic obstructive pulmonary disease treatment]. AB - The article reviews available data on direct and indirect costs of COPD treatment. PMID- 15524027 TI - [Immunomodulating properties of antibiotics]. AB - Antimicrobial agents are typically regarded as an exogenous factor responsible only for killing pathogens. However we have more and more data that numerous antibiotics have modulating influence on immunological system, not directly associated with bacteriostatic or bactericidal properties of those antibiotics. The review presents current knowledge about this issue. PMID- 15524028 TI - [Retrospective analysis of direct costs of hospital treatment of chronic obstructive pulmonary disease exacerbations]. AB - A one-year retrospective analysis of 175 cases of COPD exacerbation was conducted. A relationship was established between expenses, on the one side, age, smoking habit and pulmonary function tests results, on the other. The mean direct expenditure for hospital treatment of COPD exacerbation per person amounted to PLN 2374.9 +/- (1040.7) (PLN 1690.8 +/- 581.8 for exacerbation of mild COPD, PLN 2221.9 +/- 815.9 for exacerbation of moderate COPD and PLN 2961.4 +/- 1345.9 for exacerbation of severe COPD). A significant relationship was found between the costs of drugs and forced expiratory volume in the first second (FEV1%) and forced vital capacity (FVC%) (accordingly r = -0.19 and r = -0.26; p < or = 0.05). Smoking habit had significant influence on the costs of therapy only in men (r = 0.22; p < or = 0.05). Age did not have serious influence on the costs. The direct costs of COPD exacerbations in women were significantly higher than in men (accordingly: PLN 2723.3 +/- 1209.9 and 2206.7 +/- 906.9; p = 0.005). Pulmonary function test results, severity of disease and female sex are relevant factors influencing the direct costs of hospital treatment of COPD exacerbations. PMID- 15524030 TI - Moderating spending on prescription drugs. PMID- 15524029 TI - [Evaluation of clinical effectiveness and direct costs of azithromycin treatment for exacerbation of chronic obstructive pulmonary disease in hospitalized patients and ambulatory care]. AB - This paper presents the efficacy and cost of therapy with azithromycin for the treatment of patients with acute exacerbation of chronic obstructive pulmonary disease (COPD). Two subgroups were investigated: out-patients and hospitalized patients--20 in each group (17 women and 23 men). Azithromycin 500 mg was administered sequentially once daily. A complete physical examination including temperature, respiratory rate, cough, production and characteristics of sputum so as general aspects of quality of life assessed by COPD exacerbation questionnaire was made in hospitalized patients on a daily base and in a day 10-14 after the end of therapy. Out-patients were assessed in day 1, 3-5 days after the start of study drug treatment and 10-14 days after the end of therapy. Pulmonary function tests were assessed three times during the whole study course. The results of the study suggest similar duration of therapy with azithromycin in both study subgroups, whereas in out-patients decrease and regression of symptoms were statistically significantly quicker with tendency approximately the same in both study subgroups. The cost of therapy with azithromycin was similar in both subgroups but the complete cost of COPD exacerbation treatment was significantly lower in out-patients in comparison to hospitalized patients group (473.71 PLN and 2587.87 respectively). PMID- 15524031 TI - Healthcare joint ventures a CFO primer. AB - Joint ventures can spread costs across the partners and bring in cash. Partners should be chosen carefully, taking into account expertise and resources offered, reputation, and benefits to be gained from the deal. Even after the joint venture is in place, it needs to be monitored regularly. PMID- 15524032 TI - 3 steps for optimizing self-pay outsourcing. AB - Just because you outsource your self-pay receivables doesn't mean a dramatic improvement in return is inevitable. For optimal results, providers need to be actively involved in the process by carefully screening the vendor, coordinating activities to allow for a seamless transition, and implementing a system that accurately and continually tracks results. PMID- 15524033 TI - Job: security. 7 steps for HIPAA compliance. AB - Healthcare providers trying to meet the April 21 deadline to comply with HIPAA's security regulations need to take several actions: Study the security regulations to determine adequate levels of security for each organization. Establish a security committee and appoint an information security officer. Identify existing security measures within the organization, including administrative, physical, and technical safeguards. Conduct a thorough risk analysis. Develop and implement remediation plans. Adequately train the workforce. Periodically review these compliance activities. PMID- 15524034 TI - Tightrope walk: setting defensible prices. AB - Resolving the conflict between generating a profit and keeping prices reasonable is critical to the financial welfare of the hospital industry. Four steps may make this resolution easier: (1) determine the level of profit required, (2) assess the reasonableness of current costs, (3) assess the reasonableness of current prices, and (4) negotiate more equitable payment arrangements. PMID- 15524035 TI - Pardon me: that's our money! AB - Not-for-profit healthcare providers have several options they can use to resist government attempts to apply the charitable trust doctrine to their business decisions. As a strategic defense, providers should perform internal due diligence that focuses on governing documents, financial arrangements, donations/solicitation, real estate, and charitable waste allegations. PMID- 15524036 TI - Medicare's Compare databases with a long-term care mission. AB - Developed primarily for consumers, Medicare's Compare web sites permit immediate access to information about most nursing homes and home health agencies, including indicators of the quality of care they provide. But hospital staff, too -most notably strategic and discharge planners--can benefit by consulting Compare. PMID- 15524037 TI - There's no profiting from a joint venture misadventure. AB - In St. David's vs. IRS, a not-for-profit health system effectively challenged the IRS's determination that the system should be disqualified from tax exemption because it had entered a 50/50 joint venture with a for-profit system. The court decisions in St. David's, coupled with a recent IRS ruling, Revenue Ruling 2004 51, provide insight into how a not-for-profit hospital can structure such a joint venture to avoid jeopardizing its tax-exempt status. PMID- 15524039 TI - Medical necessity denials: prevention pays off. PMID- 15524038 TI - Raising the performance of owned physician practices. AB - A hospital-based medical group in the Pacific Northwest used a simple, recast income statement to illuminate its challenges, charged its physician leadership with overseeing the change process, and relied on supportive health system governance to provide the financing. Within 18 months, the group saw significant results in terms of both quantity and quality. PMID- 15524040 TI - 5 reasons CFOs get fired. PMID- 15524041 TI - Management's role in IT project failures. PMID- 15524042 TI - Progress review: essential to keeping your plan on track. PMID- 15524043 TI - [The role of high throughput screening in the early stage of drug research]. AB - High throughput screening (HTS) became an integral part of the early phase drug discovery process. Although the methodology itself is widely accepted as a powerful tool of lead discovery there are a number of positive and also negative opinions associated to its application. This paper gives a short introduction to HTS technology, summarizes primary experiences and points out latest changes in screening strategy and technology. Development and application of the new screening facility of Gedeon Richter is also reported. PMID- 15524044 TI - [Synthesis of hydrazino alcohols with anti-inflammatory activity]. AB - Two-step transformations (N-nitrosation and subsequent LiAlH4 reduction) of alicyclic or acyclic amines and 1,2-amino alcohols containing a secondary amino group were applied to prepare novel N1-substituted hydrazines and hydrazino alcohols with wide structural diversity. Methods for the synthesis of certain enantiopure hydrazino alcohols were also developed. The prepared compounds specifically inhibited Vascular Adhesion Protein-1 (VAP-1), a human endothelial cell adhesion molecule with a well-documented role in inflammation. VAP-1 is a semicarbazide-sensitive amine oxidase, activity of which has been demonstrated to play a role in VAP-1 induced inflammation. Some of the hydrazino alcohols obtained reduced the clinical symptoms of inflammation in experimental arthritis in rodents and appear to be potential novel anti-inflammatory drugs. PMID- 15524045 TI - [Synthesis and NMR structure elucidation of bromocryptine stereoisomers]. AB - We describe for the first time the synthesis and NMR structural characterization of two important but so far ignored diasteroisomers of bromokryptine. During this work we have made some interesting insights regarding the conformational and epimerizational behaviour of these molecules. PMID- 15524046 TI - [From routine acylation towards stable sigma-complexes of pyrimidine: carbon protonation of the pyrimdine-ring]. AB - It is well known, that the ring nitrogen of pyrimidine possesses basic property, electron donating groups increase basicity. According to literature data published so far pyrimidines protonate at the ring nitrogen. The present paper gives brief account of the C(5) carbon-protonation observed among triaminopyrimidine derivatives, which in some cases results in stable sigma complexes. Steric and electronic effects responsible for carbon-protonation are investigated in simple C(5) substituted 2,4,6-triaminopyrimidine derivatives. We show new stable sigma-complexes in the triaminopyrimidine series. The paper summarizes, in Hungarian, our recently published results (J. Amer. Chem. Soc., 125, 2535-2540, 2003) as well as our new results presented in the symposium ,,Gyogyszerkemiai Gyogyszertechnologiai Szimpozium 2003". PMID- 15524047 TI - [Amorphization in pharmaceutical technology]. AB - The amorphization of crystalline active ingredients may be necessary because of the polymorphism of the active substance, the poor water-solubility of the drug material, difficult processing in the crystalline form and the taking out of a patent for a new (amorphous) form. This article introduces protocols for amorphization, which use methods traditionally applied in pharmaceutical technology. The protocols involve three possible routes: solvent methods, hot melt technologies and milling procedures. With this presentation, the authors suggest help for practising experts to find the correct amorphization method. PMID- 15524048 TI - [Formulation of a capsule preparation with high active ingredient content using re-crystallized drug compound]. AB - Richter Gedeon Company aimed to manufacture and launch a generic product, that's original is already on market in Hungary. The goal of a generic development is to produce an essentially similar product to the original one. It seems to be simple and routine work, but during the formulation a couple of problems have to be solved. Our active agent was voluminous therefore we could not fill the whole quantity of the active, even without any excipients. In addition the flow property of the powder mixture were very bad, and it may cause inaccurate capsule filling and significant differences in the weight of the filled capsules. During the formulation we managed to assure the appropriate physical properties of the agent by re-crystallisation. We changed the excipients used in the first trials. Chemically these were the same, but with different physical features. PMID- 15524049 TI - [Investigation of redox homeostasis of liver in experimental and human studies]. AB - Great importance has been attributed to antioxidants in the treatment of conditions associated with oxidative stress for many years. At the same time the antioxidants can exert prooxidant activity. Combined antioxidant treatment is more favourable compared with monotherapy, because antioxidants have scavenger-, compartment- and tissue-specificity and they regenerate each other directly, too. Drugs of chronic liver diseases should be considered because of the role of liver in biotransformation. Our aim was to study the redox status of liver and investigate the effects of natural antioxidants (vegetable lyophilizates, silymarin, metadoxine) on liver redox status. Besides of clinical diagnosis, biochemical, analytical, histological methods were used to assess the free radical-antioxidant balance on the whole. Our in vitro methodological system is suitable for screening natural compounds for primary and secondary antioxidant property as well as membrane-stabilizing activity. A shift in free radical antioxidant balance was proved in short-term extrahepatic cholestasis, antioxidant-type choleretic compounds can be investigated in this animal model. In human experiments redox status was investigated in the Gilbert's disease and in the most frequent chronic liver disorder in Hungary, in alcoholic liver disease. The improvement of non-enzymatic antioxidant defence system was found in Gilbert's disease without the alteration of free radical-antioxidant balance. Our results proved the role of free radical reactions in alcoholic liver disease and underlined the greater women vulnerability to alcohol toxicity. The chemiluminometric measurements in erythrocytes is suitable for the estimation of progression of alcoholic liver disease and of efficacy of antioxidant therapy. PMID- 15524050 TI - [Late onset Wilson's disease]. AB - BACKGROUND: Wilson's disease (W.D.) is a metabolic disorder that occurs predominantly in children, adolescents, young adults and, rarely, in patients over 35 years. AIMS: In order to verify the prevalence of the clinical presentation of W.D. in adulthood, we analyzed a wide number of clinical presentation of W.D. with particular attention to the age of onset and to the evolutive stage of liver disease at presentation. PATIENTS: Our study is relative to 11 Sardinian adult subjects, aged 36-57 years, 6 males and 5 females, selected from a series of 120 patients affected by W.D. The only criterion utilized to select the patients was their age at presentation, with a cut off of 35 years. METHODS: Liver biopsies were routinely processed and stained with rhodanine, rubeanic acid, orcein and Timm's methods. On the basis of the histological picture, liver biopsies were subdivided into four evolutive stages: stage I = steatosis; stage II = interface hepatitis; stage III = bridging fibrosis; stage IV = cirrhosis. Molecular characterization of W.D. for gene mutations in the Sardinian population was performed in 7 out of 11 cases (-441/-427 del, 5' UTR and 3436 G > A Exon 16). RESULTS: 3 patients showed histological features of the first evolutive stage, 2 of the second, 1 of the third, and 5 of the fourth stage. Histochemistry for copper resulted positive in 9 of 11 cases at least with one of the four employed methods. In the seven patients in whom molecular characterization was perfomed, the gene mutation of W.D. was the same observed more frequently in the Sardinian population (-441/-427 del, 5'UTR). CONCLUSIONS: Our data show that: 1) W.D. with late onset is not rare in Sardinian population; 2) in spite of the late clinical presentation, W.D. may present in the first (3/10) and in the second evolutive stage (2/10) with mild to moderate changes of the liver architecture; 3) patients may show, at presentation, a severe liver disease, characterized by bridging fibrosis or cirrhosis; 4) from a practical point of view, we ask to consider the diagnosis the W.D. in all patients with chronic liver disease of unknown etiology, even if aged over 35 years. PMID- 15524051 TI - [Galanin immunoreactivity in a laryngeal paraganglioma: case report and literature review]. AB - The authors describe a case of laryngeal paraganglioma (LP) occurring in a 57 year-old-woman. To date, 70 cases have been described in the literature. It is benign and recurrences are infrequent. The differential diagnosis with typical and atypical carcinoids, hemangiopericytomas, alveolar soft-part sarcomas, medullary thyroid carcinomas, malignant melanomas and metastatic renal cell carcinomas is supported by immunohistochemistry. Moreover, this tumor shows the immunohistochemical expression of galanin, a variably expressed marker of paragangliomas that it is not expressed in carcinoid tumors. Nevertheless, our observations militate against its role as a solitary marker but advocate its use in conjunction with other antibodies for the differential diagnosis of neuroendocrine neoplasms of larynx. PMID- 15524052 TI - Colonic adenocarcinoma and bilateral malignant ovarian sex cord tumor with annular tubules in Peutz-Jeghers syndrome. AB - Peutz-Jeghers syndrome is characterized by multiple polyps throughout the gastrointestinal tract in association with mucocutaneous pigmentation. Although Peutz-Jeghers syndrome polyps are hamartomas, frequent association of this syndrome with both gastrointestinal and non-gastrointestinal tumours had led to reassessment of the cancer risk in this hereditary disorder. The most common gynaecological tumors in this syndrome are adenoma malignum of the uterine cervix and ovarian sex cord tumor, particularly sex cord tumor with annular tubules. The question of malignant change in a polyp or of the association of gastro intestinal carcinomas still discuss. The authors report a case of Peutz-Jeghers syndrome in a young patient who developed a colonic adenocarcinoma in a hamartomatous polyp together with an incidentally discovered bilateral malignant sex cord tumours. We discuss its association with certain benign and malignant tumors and the risk of rare complications of these hamartomatous polyps. Although malignant tumors are increasingly reported in association with the Peutz-Jeghers syndrome, to our knowledge, there have been no previous reports of such an association in the literature. PMID- 15524053 TI - Tumoral, quasitumoral and pseudotumoral lesions of the superficial and somatic soft tissue: new entities and new variants of old entities recorded during the last 25 years. Part II: Excerpta I. PMID- 15524054 TI - Complications of the nephrotic syndrome and their treatment. AB - The nephrotic syndrome occurs in association with a diverse array of primary and secondary glomerular disorders. Despite the different etiologies, many of the clinical effects are similar. This review focuses on the pathogenesis and treatment of edema formation, hyperlipidemia, and the hypercoagulable state. Major abnormalities of the endocrine system and evidence of erythropoietin deficiency will be reviewed. Finally, non-specific treatments aimed at reducing proteinuria will also be discussed. PMID- 15524055 TI - Examining chronic kidney disease management in a single center. AB - AIMS: The management of patients with chronic kidney disease in outpatient clinics was assessed for the ability to achieve targets of care advocated in clinical practice guidelines. METHODS: 272 records of outpatients with increased serum creatinine (> or = 1.5 mg/dl for women, > or = 2.0 mg/dl for men) were reviewed for details of their assessment and management. Prevailing data on blood pressure, anemia, bone disease and lipid status as well as therapeutic changes were evaluated. RESULTS: The subjects were aged 64 +/- 18 years, serum creatinine 2.6 +/- 1.1 mg/dl, and calculated GFR (MDRD formula) 19.2 +/- 9.9 ml/min. Median UproV was 1.0 (0.024 - 12.4) g/day. Causes of CKD were diabetes (33.5%), HTN (8.8%), GN (19.5%), and adult PKD (3.3%). Treatment targets were BP < 130/85 mmHg, Hct > or = 36%, serum Ca++ > or = 8.5 mg/dl, serum Po4 < 4.5 mg/dl and cholesterol < 200 mg/dl. Of the patients with abnormal findings, mean values for SBP were 153 +/- 17 mmHg, DBP 93 +/- 6 mmHg, Hct 31.7 +/- 2.9%, Ca++ 8.0 +/- 0.7 mg/dl, PO4 5.6 +/- 1.0 mg/dl, and cholesterol 236 +/- 37 mg/dl. Only a minority of patients with abnormal values had their treatment altered. Furthermore, only 54% of patients with hypertension were treated with either ACEi or ARB therapy. Finally, only 6% of patients with hypercholesterolemia had fasting lipid levels measured. CONCLUSION: This data suggests that treatment of patients with CKD has improved, but that many opportunities exist to optimize their care. PMID- 15524056 TI - Clinical value of renal biopsy in patients with asymptomatic microscopic hematuria with and without low-grade proteinuria. AB - BACKGROUND: The decision whether to perform renal biopsy on patients with persistent asymptomatic microscopic hematuria (AMH) with and without low-grade proteinuria (LGP) remains controversial as, although often diagnostic, the information gained seldom alters clinical management. Our study investigates the clinical value of renal biopsy in patients with isolated AMH versus those with AMH and LGP. METHODS: Between 1996 and 2002, we identified 89 patients with AMH and 46 with AMH and LGP. The patients were asymptomatic, free from systemic illness, had a sterile urine, normal serum creatinine, normal renal and bladder ultrasound, less than 2.5 g proteinuria/day, underwent successful renal biopsy and were followed-up for a mean period of 46 +/- 12 months. RESULTS: In patients with isolated AMH, thin basement membrane nephropathy (TBMN) was diagnosed in 43%, IgA nephropathy in 20%, minor abnormalities in 19% and normal biopsies in 18%. In patients with AMH and LGP, IgA nephropathy was diagnosed in 46%, other major nephropathies in 26%, minor abnormalities in 17%, TBMN in 7% and normal biopsies in 4%. At follow-up, 32% of AMH patients and 38% of AMH with LGP patients had a GFR of less than 90 ml/min and 36% and 56%, respectively were hypertensive. CONCLUSIONS: The results support the current consensus that routine renal biopsy is not indicated for isolated AMH but suggest that biopsy is indicated for AMH and LGP identifying major and potentially progressive nephropathies in 70% of patients, who should be managed by specialist nephrologists. PMID- 15524057 TI - The potential role of statins in contrast nephropathy. AB - BACKGROUND: Administration of contrast agents may result in an acute reduction in renal function and occasionally end-stage renal disease. Risk factors for contrast-induced nephropathy (CN) are preexisting renal dysfunction, diabetes and reduced effective arterial volume. Hydration and use of nonionic contrast agents have been reported to ameliorate CN. Reactive oxygen species may have a role in the pathogenesis of CN. Statins decrease free oxygen radicals in animals. We retrospectively tested the hypothesis that administering statins prior to cardiac catheterization decreases the incidence of CN. METHODS: A total of 1,002 patients were studied. Patients with a stable baseline serum creatinine (SCr) > or = 1.5 mg/dl who had cardiac catheterization between July 1997 and June 2002, were included in the study. None of the patients were taking statins before admission. 250 patients were started on a statin before the procedure and 752 patients were not. The SCr was followed for 7 days after the procedure looking for an acute decrement in renal function, dialysis requirement and survival. RESULTS: The baseline characteristics, SCr, GFR, amount of intravenous fluids and contrast were similar in both groups. The post cath SCr (2.26 vs 3.1 mg/dl, p = 0.001) was significantly better in the statin group. Length of stay (2.72 vs 3.32 days, p = 0.01) and number of patients with acute renal failure (43 (17.2%) vs 168 (22.3%) patients, p = 0.028) were significantly lower in the statin group. Dialysis requirement within 7 days and 28-day survival were similar in both groups. CONCLUSION: Prophylactic administration of statins along with hydration may be associated with less CN induced by a nonionic, low-osmolality contrast. PMID- 15524058 TI - Renal allograft glomerulopathy and the value of immunohistochemistry. AB - Studies of late renal allograft biopsies focus on chronic damage investigated by light microscopy (LM). We evaluated the use of immunohistochemistry (IH) as applied in the routine study of transplant glomerulopathies. Among renal transplants in 1985 - 1997, 129 were identified where a graft biopsy had been obtained 6 months or more after transplantation, studied by LM and IH and the original renal disease was known. IH results were evaluated in relation to glomerular LM findings and the original diagnosis. The risk of graft loss in relation to recurrent and de novo glomerulopathy was evaluated. By LM, 69 biopsies (53%) showed glomerulopathy, mesangial sclerosis only in 26, proliferative changes in 15, membranous in 15 and combined membranous and proliferative in 13. By IH, 46 biopsies (36%) stained positive with IgM and/or complement only and 24 with immune complexes including IgA and/or IgG. Seven biopsies (5.4%) showed glomerular disease by IH in spite of normal LM. Recurrence was diagnosed in 22 grafts; 12 had IgA nephropathy, 3 had SLE, 6 other immune complex nephritides and 1 systemic vasculitis. Twenty-eight biopsies (22%) with proliferative and/or membranous glomerulopathy lacked clear connection to the original renal disorder. More than half of these had deposits of IgM and C3 only. The further graft survival was significantly reduced in the presence of de novo glomerulopathy by LM, relative risk 2.0 (confidence interval 1.1 - 3.8) in a Cox proportional hazards analysis also including serum creatinine and Banff chronic allograft nephropathy (CAN) grade, p = 0.03. In conclusion, transplant glomerulopathy should be separated from recurrence. De novo glomerulopathy is frequent and ominous. PMID- 15524060 TI - Low dialysance of asymmetric dimethylarginine (ADMA)--in vivo and in vitro evidence of significant protein binding. AB - BACKGROUND: Increased blood levels of the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA) predict cardiovascular mortality in patients with end-stage renal disease. Despite its low molecular weight, available information on the impact of hemodialysis (HD) on ADMA plasma levels is controversial. METHODS: We assessed plasma concentrations, dialyzer clearance and total amount of ADMA removed in 30 patients with end-stage renal disease during regular HD. In addition, plasma ADMA levels were assessed in 10 patients with acute renal failure treated with extended HD. RESULTS: Regular HD decreased plasma creatinine (from 774 +/- 42 to 312 +/- 17 micromol/l) and urea (from 24.5 +/- 1.5 to 8.4 +/- 0.5 mmol/l) concentrations significantly (both p < 0.001), whereas plasma ADMA remained unchanged (4.35 +/- 0.19 vs. 4.76 +/- 0.24 micromol/l). ADMA clearance was 92 +/- 6 ml/min, and the total amount removed in the spent dialysate was 37 +/- 4 micromol. The clearances of creatinine (161 +/- 3 ml/min) and of urea (173 +/- 3 ml/min) were significantly higher. Furthermore, even during extended HD, plasma ADMA concentrations did not decrease significantly (1.73 +/- 0.22 vs. 1.63 +/- 0.18 micromol/l). CONCLUSION: In conclusion, dialysance of ADMA is markedly lower than expected from its molecular weight because of significant protein binding of the substance. Since markedly increased ADMA blood concentrations have been linked to cardiovascular complications due to atherosclerosis in patients with ESRD, new strategies should be evaluated to remove this putative uremic toxin. PMID- 15524059 TI - Lipid and apoprotein changes during atorvastatin up-titration in hemodialysis patients with hypercholesterolemia: a placebo-controlled study. AB - BACKGROUND: Patients with end-stage renal disease commonly present with an atherogenic lipid profile characterized by the accumulation of triglyceride-rich, apoprotein B-containing "remnant" lipoproteins, small dense low-density lipoprotein, and low levels of high-density lipoprotein. They are at increased cardiovascular risk and may benefit from drastic lipid-lowering treatment with atorvastatin, a potent, broadacting lipid regulator. This study aims to assess the effects of atorvastatin on the lipid profile in hemodialysis patients, to determine wether atorvastatin is also effective at lowering lipid levels in this particular high-risk subgroup. METHODS: In this randomized, placebo-controlled, double-blind study in hemodialysis patients with hypercholesterolemia (n = 42, mean total cholesterol 243 +/- 33 mg/dl (6.3 +/- 0.8 mmol/l)), the efficacy of 4 weekly increasing doses of atorvastatin (10 - 40 mg daily) was investigated. Lipids and apoproteins were measured in plasma and isolated lipoprotein fractions. RESULTS: Mean total cholesterol and low-density lipoprotein cholesterol progressively decreased with increasing doses of atorvastatin (total cholesterol -33%, low-density lipoprotein cholesterol -43% after 12 weeks), while high-density lipoprotein cholesterol remained unchanged. Plasma levels of apoprotein B and apoprotein E were also significantly reduced by atorvastatin 10 mg, while up-titration to 20 and 40 mg daily provided additional benefits by lowering triglycerides and apoprotein C-III. At week 12, the fraction of small dense low-density lipoprotein was significantly reduced from 23% - 18%, and apoprotein B-containing intermediate-density lipoproteins were no longer detectable. CONCLUSION: In conclusion, atorvastatin not only treated hypercholesterolemia but also favorably affected the uremic lipid profile in patients on hemodialysis. Atorvastatin 4-weekly dose escalation up to 40 mg daily was well-tolerated. Further prospective studies are needed to evaluate the impact of this improved lipid profile on morbidity and mortality. PMID- 15524061 TI - Immediate hemodialysis after percutaneous transvenous angioplasty increases patency rates of arterio-venous fistula. AB - BACKGROUND: It was reported that pathogenesis of access failure of hemodialysis patients through progressive stenosis was followed by thrombosis. Nonionic contrast media increase platelet degranulation within an angioplasty-damaged vessel by releasing procoagulant molecules, which might contribute to acute thrombosis and restenosis. An adequate level of heparin provides satisfactory thrombin inhibition during routine angioplasty. This study was performed to evaluate the effect of immediate hemodialysis after percutaneous transvenous angioplasty (PTA) to remove nonionic contrast media or other factors while injecting heparin continuously. METHODS: From September 9, 1998 - May 15, 2002, successful PTAs were performed in 66 patients with arteriovenous fistula who were not given any inhibitors of platelet aggregation. Hemodialysis was performed in 31 cases immediately after PTA, and in the remainder, hemodialysis was performed the next day. Patients were randomized and fistula patency rates were compared in these 2 groups on March 20, 2004. RESULTS: The patency rates after PTA in patients who were dialyzed immediately, were significantly higher than those who were dialyzed the next day (p = 0.0120). CONCLUSIONS: Immediate hemodialysis after PTA is an effective way of increasing the patency of arteriovenous fistula for reasons which are not clear. This observation will need to be corroborated in subsequent studies using a larger sample size. PMID- 15524062 TI - A boy with Japanese Dent's disease exhibiting abnormal calcium metabolism and osseous disorder of the spine: defective megalin expression at the brushborder of renal proximal tubules. AB - We encountered a 16-year-old boy with Japanese Dent's disease who exhibited renal insufficiency and an osseous disorder of the spine. Proteinuria first was noted at the age of 2 years. At 13 years, the patient underwent analysis of the CLCN5 gene, which identified missense mutation (I524K) in exon 10. During follow-up, a marked increase in urinary beta2-microglobulin was associated with mild deterioration of renal function. At the age of 15 years, hypocalcemia (7.5 mg/dl) accompanied by an increased plasma concentration of alkaline phosphatase was first detected. At that time, plasma concentration of 25(OH)D3 and 1'alpha25(OH)2D3 were low accompanied by a high plasma parathyroid hormone concentration. A renal biopsy specimen revealed tubulointerstitial alterations including mononuclear cell infiltration, partial fibrosis and focal glomerular sclerosis. Immunofluorescence revealed weak, discontinuous staining of megalin along the brushborder of renal proximal tubules. Western blotting demonstrated decreased urinary excretion of megalin. Thus, clinical manifestations and prognosis may vary in Japanese Dent's disease. Reduced megalin expression may have disturbed calcium homeostasis, leading to osseous disorder in our patient. PMID- 15524063 TI - Discordant phenotypic expression of Alport syndrome in monozygotic twins. AB - BACKGROUND: Alport syndrome is a genetically heterogeneous disorder, but most patients showed the X-linked form resulting from mutations in the COL4A5 gene. A few cases of mosaicism in Alport syndrome have been reported. METHODS: We describe the case of an 8-year-old boy with mosaicism in Alport syndrome. Punch skin biopsies were obtained from the patient's mother and monozygotic twin brother. Five biopsy specimens from non-Alport patients were used as controls. Immunohistochemical analysis was performed using rat monoclonal antibodies towards individual collagen IV(NC) domains. RESULTS: Kidney tissue of the patient showed: mosaic expression of alpha3(IV), alpha4(IV) and alpha5(IV) in the glomerular basement membrane (GBM), distal tubular basement membrane (TBM) and Bowman's capsule; mosaic alpha6(IV) expression in the Bowman's capsule and distal TBM; and well-preserved expression of alpha1(IV) and alpha2(IV). The patient's skin exhibited mosaic alpha5(IV) expression. His mother and monozygotic twin brother disclosed a normal linear staining of alpha5(IV) in their epidermal basement membranes. This unusual mosaicism of alpha3(IV), alpha4(IV), alpha5(IV) and alpha6(IV) is consistent with a pattern of female heterozygotes of Alport syndrome. CONCLUSION: This discordant phenotypic expression of Alport syndrome in monozygotic twins with unaffected parents suggests possible somatic mosaicism in the COL4A5 gene. PMID- 15524064 TI - Symptomatic cytomegalovirus infection complicating treatment of acute systemic vasculitis. AB - Cytomegalovirus (CMV) is usually a complication of renal/solid organ or bone marrow transplantation. We describe three cases of severe CMV in the context of vasculitis immunosuppression. PMID- 15524065 TI - Reply to Sjostrom et al. Cystatin C and creatinine for assessment of kidney function: differences in pharmacokinetics--and more! PMID- 15524066 TI - Simplest and real-time screening method of hemodialysis access recirculation. PMID- 15524067 TI - Assessment and treatment of patients with malignant spinal cord compression. AB - Spinal cord compression is a complication of malignancy that affects the quality of life of 12,700 new patients each year and disrupts the lives of the families striving to care for them. Compression can be prevented by early diagnosis, which requires a high index of suspicion on the part of patients, their families, and their clinicians. Disability arising from delays is associated with shortened patient survival. Magnetic resonance imaging is the gold standard for diagnosis and is needed in any cancer patient presenting with new back pain, whether or not plain films or bone scans show metastases. Symptomatic therapy addresses pain, constipation, spinal instability, and the psychological and social consequences of the associated disability. High-dose corticosteroids are recommended unless they are contraindicated or the patient is ambulatory and asymptomatic while receiving radiation therapy. Evidence-based guidelines recommend radiation therapy for most patients. Short courses of irradiation and reirradiation may be associated with less toxicity than previously thought. Initial surgery is recommended for patients without a previous cancer diagnosis or with a remote cancer, unstable spine or bony cord compression, or inability to receive further irradiation. New surgical data suggest that patients with irradiation-resistant tumors and a single site of compression may have improved function with initial surgery and reconstruction followed by irradiation,compared with irradiation alone. PMID- 15524069 TI - Patient-reported fatigue scores reflect physical function impairment. PMID- 15524068 TI - Treatment of brain metastases. AB - Brain metastases are a common complication of cancer, found in approximately 20% of patients at autopsy. The diagnosis is usually established by neuroimaging and carries a poor overall prognosis. Supportive therapies, such as corticosteroids, anticonvulsants, and anticoagulants, are necessary for most patients to address the common medical complications that often accompany brain metastases. These treatments often ameliorate symptoms and signs and improve neurologic function, but they require careful management to minimize their common toxicities. Definitive antitumor treatment may include whole-brain radiotherapy, surgery, stereotactic radiosurgery, and chemotherapy. A multimodal approach can yield prolonged survival of a year or more in some patients, particularly those with limited intracranial disease, high performance status, limited systemic cancer burden, young age, and certain tumor pathologies. However, even patients with poor prognostic factors can have some relief of neurologic symptoms and signs with the institution of therapy. Patients with recurrent brain metastases can also benefit from additional treatment, including all the modalities available at diagnosis. PMID- 15524070 TI - Complementary therapies for cancer-related symptoms. AB - Relief of cancer-related symptoms is essential in the supportive and palliative care of cancer patients. Complementary therapies such as acupuncture, mind-body techniques, and massage therapy can help when conventional treatment does not bring satisfactory relief or causes undesirable side effects. Controlled clinical trials show that acupuncture and hypnotherapy can reduce pain and nausea. Meditation, relaxation therapy, music therapy, and massage mitigate anxiety and distress. Pilot studies suggest that complementary therapies may treat xerostomia, hot flashes, and fatigue. Botanicals or dietary supplements are popular but often problematic. Concurrent use of herbal products with mainstream medical treatment should be discouraged. PMID- 15524071 TI - Four initiatives proposed to improve safety reporting in clinical trials. PMID- 15524072 TI - New software to facilitate clinical trials safety reporting. PMID- 15524073 TI - Hypoxia-mediated chronic normal tissue injury. PMID- 15524074 TI - Late effects of ionizing irradiation in the lung. PMID- 15524075 TI - Palliation of soft tissue cancer pain with radiofrequency ablation. AB - The purpose of this study was to analyze the feasibility, safety, and efficacy of radiofrequency ablation (RFA) to treat pain from soft tissue neoplasms. RFA was performed on 15 painful soft tissue tumors in 14 patients. Tumors varied in histology and location and ranged in size from 2 to 20 cm. Patient pain was assessed using the Brief Pain Inventory (BPI) at baseline and 1 day, 1 week, 1 month, and 3 months post RFA. All patients had unresectable tumors or were poor operative candidates whose pain was poorly controlled by conventional treatment methods. BPI scores were divided into two categories: pain severity and interference of pain. Although not all scores were statistically significant, all mean scores trended down with increased time post ablation. Based on these outcomes, RFA appears to be a low-risk and well-tolerated procedure for pain palliation in patients with unresectable, painful soft tissue neoplasms. RFA is effective for short-term local pain control and may provide another option for failed chemotherapy or radiation therapy in patients with cancer. However, pain may transiently worsen, and relief is often temporary. PMID- 15524076 TI - Use of atypical antipsychotic agents for symptom control in patients with advanced cancer. PMID- 15524077 TI - Baseline patient factors predict response to erythropoietic therapy. PMID- 15524078 TI - Silence as the speech of care. PMID- 15524079 TI - Higher costs may justify prophylaxis in prostate and lung cancer patients with bone metastases. PMID- 15524080 TI - [Health beliefs and health behavior in older employees of medium-size and large enterprises]. AB - BACKGROUND: In view of the aging process in our population there is a need to identify effective methods to prevent disadvantageous effects of this trend. In this paper the state of particular health-related behaviors and elements of health awareness among retirement-approaching employees are presented as a basis for developing efficient health promotion programs for this age group. MATERIALS AND METHODS: The data presented in this paper have been gathered during the study conducted on a random sample of 1138 employees of two 25-35 and 45-55 age groups. The study was carried out in the form of a questionnaire-based interview. RESULTS: There is no significant difference between older and younger employees with regard to their attitude towards one's own health. There are two major problems common to both age groups, the lack of awareness of very close relationship between everyday behavior and health, and a low locus of control manifested by finding excuse for health negligence in numerous problems faced each day or lack of strong will. The most specific feature in older workers is that they concentrate their health-related thinking and acting on the use of services provided by medical specialists. CONCLUSIONS: There are no sound grounds to justify the development of health promotion programs addressed only to older employees. The effective solution to this problem could be the introduction of workplace health promotion programs focused on enhancing the workers' sense of responsibility for their health and on developing skills to shape conditions favorable to health conducive behaviors. PMID- 15524081 TI - [Effect of chronic lead poisoning on the parameters of heart rate variability]. AB - BACKGROUND: The dysfunction of the autonomic nervous system is one of the effects of chronic exposure to lead. Heart rate variability (HRV) is a non-invasive method used to estimate the autonomic system. The aim of the study was to estimate HRV in people occupationally exposed to lead. MATERIALS AND METHODS: The study group included 43 copper-smelters and 43 healthy subjects matched individually. A 24-hour ECG Holter monitoring was performed in all men. RESULTS: The HRV time analysis in a 24-hour observation, as well as for daily activities (6:00-22:00) and nocturnal sleep (22:00-6:00) showed that pNN50 parameter, an estimator of the vagus nerve function, was significantly lower in copper-smelters than in healthy subjects (p < 0.05). The majority of time and spectral HRV parameters, estimated at 15-min intervals during day- and night-time in the group of lead exposed men, were lower than in the control group (p < 0.05). CONCLUSIONS: In subjects occupationally exposed to lead, HRV is lower than in healthy men, which results rather from the decreased parasympathetic than from the increased sympathetic activity. PMID- 15524082 TI - [Influence of tobacco smoking on endothelial function in lead-exposed male workers]. AB - BACKGROUND: Endothelial dysfunction is a pathology resulting from smoking and occupational exposure to lead. The aim of the study was to examine the influence of cigarette smoking on biochemical parameters of endothelial function in people occupationally exposed to lead. MATERIALS AND METHODS: The study covered 105 men, including 43 lead exposed copper-smelters (mean age 49.9 +/- 4.1) and 62 non exposed men (mean age 47.3 +/- 9.3) formed the control group. In all subjects, peripheral blood lipid concentrations, biochemical parameters of endothelial function (nitric oxide, endothelin-1, sICAM-1, selectin-E) and inflammation parameters (interleukin-1beta, interleukin-6) were measured. RESULTS: In the group of smokers, including both copper-smelters and control subjects, serum nitric oxide concentrations were lower than in non-smokers. In the serum of men occupationally exposed to lead, sICAM-1 concentrations were higher than in the control group. The lack of physiological correlation between serum concentration of nitric oxide and endothelin-1 resulted from interactions of tobacco smoking with lead exposure. CONCLUSIONS: Workers occupationally exposed to lead should be informed about adverse effects of tobacco smoking in terms of the interaction between nicotine and lead, two combined toxic agents affecting the circulatory system. PMID- 15524084 TI - [Menstrual disorders. Is this a problem to be handled by occupational medicine physician?]. AB - BACKGROUND: The aim of the study was to define the frequency of menstrual disorders and identify risk factors, especially those associated with the work environment. MATERIALS AND METHODS: The study group was composed of 142 women, aged 22-45 years. Some of them were employed in a cosmetics manufacture plant and a bank, others were hospital and ambulatory nurses and auxiliary personnel. RESULTS AND CONCLUSIONS: The study revealed irregular cycles in one third and prolonged bleeding in one fourth of the women. Long cycles were observed in one fifth of the women and short cycles were noted in every tenth women. It was found that occasional or frequent contact with ethylene oxide increased the risk of irregular cycles and oligomenorrhea, whereas work around X-ray apparatus was responsible for the risk of long cycles and heavy bleeding. Long rest pauses during work (in this study correlated with work load) were regarded as a factor inducing the risk of oligomenorrhea and prolonged bleeding. A randomly sampled group of 33 women measured morning temperature during two consecutive cycles. Of this number, 15 women (45.5%) showed ovulation in both cycles, 11 (33.3%) in one cycle only, and 7 both anovulatory cycles. The risk of ovulation disorders was enhanced by a two-shift work system (mostly night shift) and strong job stress. Energy expenditure for leisure time activity > 1000 kcal/week and smoking also contributed to an enhanced risk. In summing up, it should be stressed that because of varied frequency of irregular cycles determined by the intensity of environmental factors, occupational medicine physicians should gather data on the occurrence of such disorders and undertake relevant preventive measures. PMID- 15524083 TI - [Recommended frequency of preventive psychological tests in selected groups of operators]. AB - BACKGROUND: Psychomotor efficiency, essential in the job as an operator, declines with age. The aim of the present study was to assess the dynamics of changes and to work out recommendations on the frequency of psychological test performed during preventive checkups. MATERIALS AND METHODS: Psychological test battery was performed twice at an interval of 3-5 years, in a group of 167 drivers of motor vehicle, and operators of cutting (in surface mining) and earth-digging machines, aged 25-60 years. Performance and paper-pencil tests were used to measure psychomotor and intellectual functions. RESULTS: The statistical analysis revealed significant age-related decline in attention, cognitive and vital functions and eye-hand coordination. The most advanced deterioration was observed in the group of workers aged 50 years and more. CONCLUSIONS: As revealed by the study, psychological tests should be performed in operators during all the three types of preventive checkups: (a) pre-placement in candidate workers; (b) control in case of the central nervous system impairment; and (c) periodic, carried out at least every four years in workers aged 50 years and more. PMID- 15524086 TI - [Assessment of annual exposure to noise and risk of occupational hearing loss among private farmers specializing in plant production]. AB - BACKGROUND: In agriculture noise is the most frequent hazard, but until now very poorly recognized. It affects the hearing organ in private farmers. Because of a great variety of machines used on farms and a changing magnitude of exposure to noise, this agent has to be studied over the whole annual cycle. The aim of the study was to recognize and evaluate annual exposure to noise among private farmers engaged in plant production. MATERIALS AND METHODS: The study covered 17 family farms with arable land, ranging from 10 to 50 ha (mean, 22.3 ha). They were equipped with agricultural tractors and a set of agricultural accessories, wood-cutting saws, self-propelled and workshop machines. Based on detailed time schedule records of agricultural activities and dosimetric measurements conducted during the whole year, two acoustic parameters were determined, total monthly exposure and equivalent daily exposure. RESULTS: The study showed that the highest values of the total monthly exposure to noise (E(A,T)) occurred in the summer-autumn (July, September, October) and winter (December, January) seasons. During the former, the degree of noise load among farmers was directly associated with intensity of field works and transport activities, whereas during the latter with the frequency of using wood-cutting machines and repair activities. The calculated mean equivalent daily exposure to noise revealed the highest values in two months, December (6.01 Pa2 h) and September (5.51 Pa2 h), and the lowest values were observed in the other months (3.67-4.28 Pa2 h). The mean value for the whole year was 4.35 Pa2 h (the standard was exceeded by 4.4 times). CONCLUSIONS: The study showed that the mean annual noise load among farmers engaged in plant production was characterized by the level equal to 91.3 dB-A with the highest hearing risk in December and September. In those two months, maximum allowable exposure to noise was exceeded by six times (E(A,8h)). Owing to the precision of the obtained values and their strong uniformity, they may be used in practice by relevant services to evaluate the degree of noise risk among private farmers engaged in plant production. PMID- 15524085 TI - [Effectiveness of manual therapy combined with physiotherapy in musicians' occupational overloads]. AB - BACKGROUND: Manual therapy, kinesitherapy and physiotherapy are very helpful in the treatment of musicians' occupational overloads. The aim of this work is the estimation of the effectiveness of these three methods in the therapy of instrumentalists' occupational overloads. MATERIALS AND METHODS: Examinations were made among 689 professional musicians. They were divided into three groups: functional disorders, primary organic disorders, and secondary organic disorders. Before and after the treatment some parameters of the movement organs were estimated. RESULTS: Among patients with functional disorders, after manual therapy all estimated movement organs' parameters improved, however among patients with organic disorders, bigger improvement of some parameters were noticed after kinesitherapy and physiotherapy. CONCLUSIONS: Manual therapy is effective method of treatment of movement functions disorders caused by occupational overloads. PMID- 15524087 TI - [Exposure of nurses to electromagnetic fields]. AB - BACKGROUND: Devices that produce electromagnetic fields (EMF) within the range of 0-300 GHz are widely used in surgical and diagnostic procedures. As a result a large number of physicians and other groups of medical personnel may be exposed to EMF. Even if patients' exposure, sometimes quite high, is inevitable or even recommended, medical personnel should be substantially protected against EMF exposure. MATERIALS AND METHODS: Evaluation of nurses' exposure to EMF was based on an analysis of EMF magnitudes in the surrounding of magnetic resonance imaging (MRI) and electrosurgical units. These two kinds of apparatus are the strongest EMF sources in health service facilities. The measurements were performed according to the norms and hygiene regulations binding in Poland. Measurements performed by the Nofer Institute of Medicine in Lodz, and data collected by the Central Database on EMF Sources were used in the analysis. The Central Database is run by the Nofer Institute of Medicine at the behest of the Chief Sanitary Inspector. RESULTS: The study showed that nurses' exposure to EMF emitted by MRI and electrosurgical units complies with Polish norms and hygiene regulations and can be classified as negligible or allowable. CONCLUSIONS: It was found that work of nurses in exposure to EMF emitted by MRI and electrosurgical units can be regarded as safe, which means that their health should not be endangered by performing their job. PMID- 15524088 TI - [Should ticks be regarded as a tularemia vector in habitants of North-Eastern Poland?]. AB - BACKGROUND: A growing incidence of tick borne encephalitis and Lyme borreliosis in north-eastern Poland as well as an increasing number of Ixodes ricinus ticks in wooded areas and meadows have prompted the authors to evaluate the presence of antibodies against Francisella tularensis in serum of forest workers. MATERIALS AND METHODS: Sera of 820 persons, including 765 healthy forest workers (group I) and 55 patients with lymphadenopathy (group II) admitted to the Department of Infectious and Neuroinfectious Diseases, Medical Academy, Bialystok, were diagnosed for antibodies against F. tularensis. Agglutination test at 37 degrees C, with suspensed F. tularensis (Biomed, Krakow, Poland) were used. Simultaneously all sera were tested for the presence of antibodies against B. burgdorferi, using ELISA test, Borrelia IgM and Borrelia IgG recombinant (Biomedica, Austria). RESULTS: Of the 765 forest workers (group I), 20 (2.06%), persons showed positive (1:20) result of agglutination test, including 8 persons with antibodies against B. burgdorferi in IgG, of these 1 in IgM and IgG. Twenty persons with positive agglutination test had no signs of present or past tularemia in physical examination. Nine persons with prior diagnosis of arthritic form of Lyme boreliosis had undergone repeated antibiotic therapy (two 30-days therapy courses with cefotaksym or doxycycline). In group II, sera of 55 patients were negative in the agglutination test with F. tularensis. CONCLUSIONS: The results of our study show that in North-Eastern Poland, Ixodes ricinus is not essential in Francisella tularensis transmission in this region. PMID- 15524089 TI - [Toxicogenomics. New perspectives for the molecular toxicology]. AB - Gene expression in the cellular genome is subject to changes caused by internal and external factors. Due to different expression of gene sets (polymorphism), cells show different morphological and functional characteristics. Environmental and occupational toxic agents may influence cells at the level of transcription and translation. The functional toxicogenomics attempts to explain those influences. Due to recent developments in molecular biology and bioinformatics, it has become possible to analyze protein transcript (toxicogenomics) and profile (toxicoproteomics). This work reports new opportunities to study gene sequencing and expression by means of the DNA chip technique (rapid analysis of the genetic polymorphism) and the microarrays technique (simultaneous analysis of hundreds or thousands of genes). The authors report examples of some practical applications of toxicogenomics in the assessment of the effects of pathological exposures to environmental and occupational toxic (carcinogenic, hepatotoxic and/or neurotoxic) agents, due to the development of new groups of biomarkers, such as biomarkers of individual susceptibility, biomarkers of toxic effects combined with the assay of the relationship between a toxic agent and its dose, and the effect measured at the level of the cellular genome and results of histopathological and biochemical tests. PMID- 15524090 TI - [Allergy to cosmetics. I. Fragrances]. AB - The authors report current information on allergy to aromatic agents present in cosmetics and products of household chemistry. In the perfume industry, about 3000 aromas are used. Single products may contain from 10 to 300 compounds. The problem of difficulties encountered in the diagnosis of hypersensitivity to odors is addressed. The mixture of 8 such products used in diagnostic screening is able to detect allergy only in about 30% of patients who do not tolerate cosmetics. Changing frequency of allergy to individual aromas is discussed. It has been now observed that cinnamon products are less allergic than chemical compounds present in oak moss. Since the 1990s of the last century, allergy to a synthetic aromatic agent, Lyral is the subject of interest in many research centers involved in studies of contact allergy. Half the cosmetics present in European markets, especially deodorants, after shave cosmetics, hand and body lotions contain this agent. It induces positive reactions in about 10% of patients allergic to aromatic agents. Detection of allergy to Lyral is difficult as it is not included in the set of commercial allergens used to diagnose hypersensitivity to aromatic agents. PMID- 15524091 TI - [Allergic fungal sinusitis: diagnostic, therapeutic and prognostic evaluation]. AB - Allergic Fungal Sinusitis (AFS) is a benign noninvasive sinus disease related to hypersensitivity to fungal antigen. The incidence of the disease has been estimated to be 4-7% of patients requiring surgery for chronic sinusitis. AFS is characterized by high levels of IgE in the atopic patient's serum, and the presence of allergic mucin that contains eosinophils and fungal components. The treatment of AFS entails endoscopic sinus surgery, combined with pre- and post operative systemic and local steroid treatment. This article summarizes the clinical course of 11 patients treated in our department for AFS between the years 1996 and 2002 and followed-up for more than twelve months. We review the characteristics of AFS, the diagnostic criteria, the treatment protocol and the prognostic outcome in these patients. PMID- 15524092 TI - [Fatal pancytopenia and methotrexate-trimethoprim-sulfamethoxazole interaction]. AB - Low dose methotrexate [MTX] is now frequently used for various inflammatory diseases. This is a case study of a fatal outcome in a patient with rheumatoid arthritis [RA] treated for a short period with low dose MTX. The patient developed severe pancytopenia followed by bacterial and monilial sepsis upon the co-administration of trimethoprim-sulphamethoxazole [TMP-SMX] for an intercurrent infection. The differential diagnosis of pancytopenia and the mechanisms underlying the increase in plasma free MTX by MTX-SMX in the patient are discussed. It should be noted that this fatal case highlights the risk of severe drug interactions in patients with multiple risk factors treated with low dose MTX for a short period of time. PMID- 15524093 TI - [Sarcoidosis--an uncommon cause of pancreatic mass]. AB - Sarcoidosis is a multi-system granulomatous disease that mainly affects the lungs and the lymphatic system. Involvement of the digestive system is rare, while the presentation of a mass in the pancreas as the first sign of sarcoidosis in the digestive system is yet rarer. Very few cases have been reported in the literature. The presentation of a mass in the pancreas is indicative of malignancy or inflammation and thus makes the diagnosis of sarcoidosis very difficult. Often it is discovered only after surgical intervention. We present a case study in which the involvement of the pancreas, bile ducts and mesentery lymph nodes led to a Whipple operation. However, on biopsy, non-caseating granulomas were found with no signs of malignancy. In view of the difficulties in diagnosing sarcoidosis of the digestive system and pancreas, we present a review of the literature on the diagnosis and management of this form of sarcoidosis. PMID- 15524094 TI - [Practical significance of sonographic type IIa of dysplastic hip joint according to Graf's classifications]. AB - Hip joint assessment in newborns and neonates is currently performed with ultrasonography due to the high degree of sensitivity and specificity of this technology as compared to the usual radiography. Measuring the alpha and beta angles is indicative in assessing the hip joint. The alpha angle represents the bony acetabular components and the beta angle represents the cartilaginous roof. According to Graf's classifications, type IIa is subdivided into two subgroups; type IIa- and IIa+. This study examined the need for following-up on these hip joints in newborn and neonate. The study population included the newborns and neonates born in our facility between the period 1/1/1999 to 1/4/2002. We routinely screened 10,432 newborns both clinically and ultrasonographically. This included a total of 20,862 hip joints, of which 915 were pathological according to Graf's classifications. Type IIa was diagnosed in 232 newborns and a total of 330 hip joints. The follow-up examinations were performed at 6 and 12 weeks of age, and at six months of age. Each follow-up visit included clinical and sonographic evaluation and alpha angle measuring. The incidence of type IIa was 1.6% of all hips and 36.9% of the pathological hip joints. These joints were divided into two subgroups. Group A included 254 hip joints among 156 newborns, 116 girls [74.3%] and 40 boys [25.6%]. Group A members had bilateral or unilateral type IIa hip joints. In the latter case, the other joint was type I normal. Group B included 76 hip joints among 76 newborns, 72 girls [94.5%] and four boys [5.5%]. Group B members had one type IIa hip joint and a second more severely graded hip joint. Follow-up only was required in 88% of the type IIa hip joints and 12% were treated using Pavlik's method. Surprisingly, all the joints with unilateral type IIa hips developed into normal or type I by six months of age without treatment, independent of the alpha angle value, and the pathology on the opposite side. PMID- 15524095 TI - [A long-acting second generation anti-psychotic--experience in Israel]. AB - Schizophrenia is a chronic disease characterized by psychotic symptoms as well as negative symptoms such as affective flattening, social withdrawal and occupational dysfunction. Anti-psychotic medications reduce the risk of psychotic exacerbations and hospitalization. Poor compliance is common among patients with schizophrenia. Long-acting medications have such advantages as stabilizing drug levels and improving compliance. Second generation anti-psychotic medications were found to be more effective and tolerable compared to first generation drugs. These medications cause less extra-pyramidal symptoms, and compliance with them was shown to be better. Until recently there were only first generation long acting anti-psychotics in use. Recently a new second generation long-acting anti psychotic drug was introduced in Israel. We present our experience with a first schizophrenic patient treated with long-acting Risperidone (Risperdal Consta). The patient was treated in the past with several first generation anti-psychotics and suffered severe extra-pyramidal symptoms. His compliance with treatment was poor. Under treatment with oral Risperidone a considerable improvement was recorded, however compliance remained poor. Under treatment with long-acting Risperidone, Intramuscularly 25 Mg every two week, both positive and negative symptoms improved substantially, as well as compliance with treatment. The results of this case study encourage us to believe that many more patients will benefit from the advantages of both a second-generation anti-psychotic and a long acting preparation. PMID- 15524096 TI - [Outcome of a residency program in radiology for immigrants and other physicians by Maccabi Healthcare Services]. AB - BACKGROUND: Maccabi Healthcare Services has more than 20 Diagnostic Imaging Clinics in Israel, and many radiologists are needed to perform and interpret the imaging studies. There was a shortage of radiologists willing to work full-time in these clinics. In the late 1980s and early 1990s a large number of immigrants arrived in Israel, some of them radiologists, who were looking for a position at Maccabi. However, their training was inadequate for the needs of modern radiology. Therefore, a program in radiology was initiated to support a number of these physicians to complete additional training in radiology, a process that would facilitate their employment as radiologists for Maccabi Healthcare Services. OBJECTIVES: To support several years of residency in radiology for immigrants and other physicians who would, on completion of their residency, work as radiologists for Maccabi Healthcare Services. METHODS: A total of 21 physicians entered the program, 16 women and 5 men, including 11 physicians from the former Soviet Union. They started their training in various academic hospitals during the years 1991-1997. A questionnaire regarding the program was completed by the residents and the chairmen of the relevant departments were interviewed. RESULTS: All physicians except one, who left the country for medical reasons, have as of now successfully completed their residency, and are employed as radiologists in Maccabi. One candidate received a grant for one year of residency abroad, and has been employed by Maccabi since completing her residency. CONCLUSION: The program can be termed a success, as all the parties concerned--residents, Maccabi Healthcare Services and the participating hospitals, profited from the arrangement. PMID- 15524097 TI - [On the relevance of protein kinase C to lithium therapy in bipolar disorder]. AB - The discovery of lithium's efficacy as a mood-stabilizing agent revolutionized the treatment of patients with bipolar disorder and after 5 decades this drug continues to be the mainstay of treatment of this disorder. Valproate, which is dissimilar structurally to lithium, shares most of the effects of lithium at the level of protein kinase C (PKC). Both drugs reduce the activity of PKC, though via different mechanisms. In comparison to patients with major depressive disorder, schizophrenia, or healthy controls, PKC activity is significantly elevated in manic patients, suggesting that changes of PKC activity may be a central pathological trait of the illness. The precise physiological role of PKC activity in the regulation of mood is unclear. The enzyme modulates cellular responses via phosphorylation of numerous substrate proteins. Such substrates of PKC include cytoskeletal proteins, neurotransmitter and hormone receptors, G proteins, GAP-43, MARCKS etc. Further studies are required to clarify any causal role of CPK changes in bipolar-disorder. PMID- 15524098 TI - [Female sexual dysfunction (FSD)]. AB - Recent research suggests that a large number of women are dissatisfied with their sexual life due to sexual dysfunction. The accepted definitions of female sexual dysfunction are based on the human sexual response cycle described by Masters and Johnson and later elaborated by Kaplan. Their work formed the basis for the diagnostic systems of both the ICD-10 and DSM IV. Female sexual dysfunction may appear as lack of sexual desire, sexual pain, and arousal or orgasmic dysfunction. Although these problems are very common in women and more common than in males, for many years this field of interest was neglected both scientifically and clinically. In the past, only psychological and sexological forms of therapy were suggested. It is now better understood that, as in many other areas, the junction between the body and the mind, is responsible for many sex problems in women. Furthermore, multi professional team work is needed to treat these problems in order to enhance better quality of life for individuals and couples. This editorial calls for better education and research in this important field. PMID- 15524100 TI - [Sexual dysfunction in women]. AB - BACKGROUND: Sexual dysfunction in women is quite prevalent and encompasses all ages, cultures and socio-economical statuses. The various array of sexual dysfunction in women adversely affects their quality of life on the one hand and their relationship with their spouses on the other. GOALS: To overview the issue of sexual dysfunction in women while focusing on the latest medical treatment options available. METHODS: Review of the relevant medical literature. RESULTS: Medical treatment of sexual dysfunction in women is based on medications that either enhance blood flow into the female sex organs, mediate sexual neurological response or increase female libido. Most of these treatments are still experimental, yet have demonstrated encouraging preliminary results. CONCLUSIONS: Medical treatment of female sexual dysfunction (FSD) is an evolving discipline in contemporary medicine that has recorded several successes. However, in light of its complexity, much is still left to be resolved in the complete understanding of the female sexual response and treatment should be adjusted accordingly. PMID- 15524099 TI - [Possible involvement of protein kinase C in the pathophysiology and treatment of bipolar disorder]. AB - Over the past decade, the focus of research into the pathophysiology of bipolar disorder has shifted from an interest in the biogenic amines to an emphasis on second messenger systems within cells. Emerging evidence implicates protein kinase C (PKC) intracellular signaling cascade in the pathophysiology and treatment of bipolar disorder. This review explores the possible involvement of PKC in bipolar disorder summarizing results from laboratory and clinical studies. Bipolar patients were demonstrated to have altered PKC levels, activity or distribution in platelets and in the brain. Chronic administration of lithium and valproate produced a striking reduction in protein kinase C (PKC) human cells and in rats. PKC inhibition in animals resulted in altered affective-like behavior and in a small study, tamoxifen (a PKC inhibitor) had marked antimanic efficacy. The results of studies at the molecular, cellular, animal and clinical levels all suggest that regulation of PKC signaling pathways may play a major part in the pathophysiology and treatment of bipolar disorder. Therefore, this pathway may be a promising candidate for the development of new, more specific drugs for the disease. PMID- 15524101 TI - [Androgen receptor and male infertility]. AB - The androgen receptor (AR) mediates androgen action determining male sexual phenotypes and promotion of spermatogenesis. Mutations in the AR cause various degrees of androgen resistance resulting in a range of androgen insensitivity syndromes. A single copy gene in the X chromosome encodes the AR. The gene contains a polymorphic triple repeat sequence [(CAG)n] with 9-36 repeats in the normal population, and displays ethnic dependence. In vitro, there is an inverse correlation between CAG repeat length and AR function. Associations exist between short alleles and prostate cancer in men or clinical hyperandrogenism in women. Expansion of the CAG tract > 40 repeats leads to spinal bulbar muscular atrophy (SBMA, Kennedy disease), an adult onset neurodegenerative disease that also presents with low virilization and spermatogenetic defects. The disease may show evidence of anticipation (increasing severity with succeeding generations accompanying further expansion of repeat length). Twelve studies involving Singaporean, Australian, North American and Japanese men reported a relationship between AR CAG repeat length and male infertility, whereas 10 studies, most of them European, found no association. Differences in hereditary or acquired factors in these populations may explain the equivocality. However, statistical methods, sample sizes, study definition and control populations, in addition to laboratory methods vary widely within the published papers, and could affect the results and conclusions. Current data is insufficient to conclude whether IVF patients who display AR CAG expansion may transfer infertility or premutation of neurodegenerative disease to their descendants. We recommend screening of AR CAG repeat length, at least in those populations where an association between repeat length and infertility could be found. PMID- 15524102 TI - [Trauma education of general surgery residents]. AB - Educating residents in surgery, especially in trauma surgery, and the best way to teach it among the other subspecialties of surgery, is a topic heavily debated in recent literature. Several important factors affecting this learning have changed lately. Many educators have expressed concern about the adequacy of the exposure of surgical residents to surgery and of their ability to care for their patients, with the newly imposed legal constraints on the length of shifts and working hours. Since trauma is inherently unpredictable, residents must spend many hours in the emergency department to become exposed to various types of trauma. Some will only see a small number of trauma patients during their residency, depending on the location of hospital and its commitment to trauma. Furthermore, the operative exposure in trauma has decreased dramatically, with the current trend of non-operative management of blunt splenic and hepatic trauma and conservatism in the management of some stable patients injured by penetrating mechanisms. Similar concerns have also been raised about the exposure of attending surgeons to surgical trauma practice. In most centers in Israel, the trauma patient is managed by a general surgery specialist rather than by a trauma-specialized general surgeon. This may lead to dilution of trauma cases among multiple medical providers and furthermore, to inconsistency in the delivery of care. Consequently, this may effect the education of surgical residents. We therefore suggest a structured curriculum for general surgery residents as well as structural changes in the department of surgery to increase the caseload of general surgeons especially those choosing to specialize in trauma. PMID- 15524103 TI - [Fever and its decrement in acute stroke]. AB - Management of body temperature is considered crucial in the management of acute stroke patients. Hyperthermia exacerbates ischaemic neuronal injury and physiological dysfunction. Hypothermia has been suggested as the "gold standard" for neuroprotection to which new pharmacological therapies should be compared. This article reviews the literature on the predictive role of admission body temperature on the outcomes of patients with stroke. We also examined the derived effect of using hypothermic procedures and antipyretic agents as a treatment measure in acute stroke patients for improving outcomes. Immediate effective treatment of the fever, while simultaneously evaluating its origin, is found to be crucial in the acute stroke for improving outcomes. The hypothermic effect of acetaminophen as a preventive measure for all patients requires further evaluation. PMID- 15524104 TI - [What do hospitalized mental patients complain about and how to deal with their complaints]. AB - Mental ill patient are a population at risk for human and civilian rights violations. One way of preventing this is by the institution of the Ombudsman. In this paper we present the Beer Yakov way of implementing the Ombudsman and review the results of four years of his activity. We also discuss the pros and cons of our model. In general we conclude that not only does the Ombudsman fill a role determined by law, but he also represents a very important link between therapists, patients and families, and makes a very important contribution in improving communication between all parts in the Mental Institution. PMID- 15524105 TI - [National Health Survey 2003-2004 on physical and mental health]. AB - The National Health Survey is a cross-sectional study initiated by the Ministry of Health as part of the World Mental Health 2000 Survey of the World Health Organization. The survey is being conducted in collaboration with the Central Bureau of Statistics, which is responsible for the field work and is supported by the National Insurance Institute of Israel and the Israel National Institute for Health Policy and Health Services Research. For the first time in Israel, national data will be collected on: (a) prevalence of mental disorders; (b) prevalence of impairments and disabilities; (c) associations between chronic conditions and mental disorders; (d) associations between socioeconomic and demographic characteristics, mental health, physical health, disability, use of health services and out-of-pocket medical expenditure. The survey will provide the much-needed database for the proposed national reform of mental health services. This article describes the main innovations in the national health survey. PMID- 15524106 TI - [Biosynthesis of the sulfated glycosaminoglycans in relation to human diseases]. PMID- 15524107 TI - [Physiological role of CDK inhibitors]. PMID- 15524108 TI - [Control of T cell fate by galectins and their ligand-through regulation of the expression of glycosyltransferases]. PMID- 15524109 TI - [Regulatory mechanism of SNARE-mediated exocytosis]. PMID- 15524110 TI - [Molecular mechanisms of lymphatic development]. PMID- 15524111 TI - [Crosstalk between ceramide and phosphatidylinositol-3 kinase as a mechanism for cell death determination]. PMID- 15524112 TI - [The diversity of plant ABC protein superfamily]. PMID- 15524113 TI - [Functions and evolution of siglecs, a family of endogenous sialic acid receptors of vertebrates]. PMID- 15524114 TI - Doctors are still burying their mistakes. PMID- 15524115 TI - Contaminated blood. PMID- 15524116 TI - What price imprisonment? PMID- 15524117 TI - The lessons from the Sally Clark case. PMID- 15524118 TI - An update on managing dangerous people with severe personality disorder--"the human time bombs.". PMID- 15524119 TI - Emerging ethical issues. PMID- 15524121 TI - Like a beacon in the night. PMID- 15524122 TI - Women's health: state by state. From heart disease prevention to mental health, states fall short in many areas. PMID- 15524124 TI - Center for American Nurses: serving the needs of nurses today & tomorrow strategies for a successful retirement. PMID- 15524125 TI - ASNA independent study. Fibromyalgia. PMID- 15524126 TI - Basic ground rules for Alabama nurses. PMID- 15524127 TI - [Pregnant, addicted prostitutes: compulsory admission is sometimes necessary in the interests of the child]. AB - Three drug-addicted prostitutes, who were 37, 27 and 25 weeks pregnant respectively, were admitted to a psychiatric department under court order. After the birth, the children were placed in foster families. The mothers withdrew from further treatment. In Rotterdam, The Netherlands, there is a city network made up of doctors and social workers; in the case of addicted pregnant prostitutes they advise the Child Protection Agency of the Ministry of Justice about the care of the child after its birth. The combination of heavy addiction and prostitution in a pregnancy without any prenatal care can be considered a psychiatric disorder. Consequently, an addicted woman can be compulsorily admitted because of the threat she forms to her unborn child. In these cases the mother's right to self determination must be limited in the interest of the child. PMID- 15524128 TI - [Migraine and TIAs; possibly more overlap than difference]. AB - Headache and transient neurological deficits can occur both in patients with migraine and in patients suffering a transient ischaemic attack (TIA). Consequently differentiating between these two disorders may be difficult. Recently, silent cerebral infarcts and ischaemic lesions in the subcortical white matter were found to be associated with migraine and in particular, with migraine with aura. The predominance of cerebellar infarcts in these patients is difficult to understand. Whether ischaemic damage in the brain of patients with migraine should be managed in a manner similar to that of patients with TIAs, should be the subject of future research. PMID- 15524129 TI - [Osteotomy at knee level for young patients with gonarthrosis]. AB - Young patients with gonarthrosis that does not respond adequately to conservative therapy can be treated by corrective osteotomy. Osteoarthritis of one compartment more often has a mechanical aetiology than osteoarthritis of the entire knee. Patients with osteoarthritis of the medial compartment often have a genu varum (bow-legs) while patients with osteoarthritis of the lateral compartment often have a genu valgum (knock-knees). The goal of corrective osteotomy is to transfer the load bearing to the normal compartment, which will reduce the symptoms and permit arthroplasty to be postponed. In retrospective studies, the procedure resulted in less pain, improved knee function or postponement of knee arthroplasty in 28-87% of the patients. Possible complications include pseudarthrosis, thromboembolism, contracture of the patellar tendon, paresis of the N. peroneus, compartment syndrome. The outcome of osteotomy for gonarthrosis depends on careful patient selection, the stage of osteoarthritis, and the achievement and maintenance of the correction of the load axis that was calculated before the operation. PMID- 15524130 TI - [Medial unicompartmental knee prosthesis for patients with unicompartmental gonarthrosis]. AB - The function and survival time of unicompartmental knee prostheses for patients with severe gonarthrosis have been improved the past few years by developments in their design, the instrumentarium and the surgical technique. A medial unicompartmental knee prosthesis may be indicated in patients with arthrosis of the medial tibiofemoral compartment. The prerequisites are an intact anterior cruciate ligament, an intact lateral compartment, a correctable varus axis and sufficient flexion in the knee. Contraindications are inflammatory arthropathies and a recent episode of septic arthritis. Relative contraindications are: old age, excess body weight, patellofemoral arthrosis and chondrocalcinosis. A unicompartmental knee prosthesis can be placed via a small parapatellar incision. The postoperative recovery is more rapid than following the classical open approach, while the knee function after 5 years is comparable. The knee function also seems better following a medial unicompartmental knee prosthesis than after valgusising tibial head osteotomy. The latter is still preferred for young active patients. Medial unicompartmental knee prostheses fail in 6-8% of patients. Revision to a total knee prosthesis is then the treatment of choice. In the long term, unicompartmental prostheses with a mobile bearing become loose less often than comparable prostheses with a fixed bearing. PMID- 15524131 TI - [Diagnostic image (209). A man after submersion in water]. AB - A 32-year-old man was presented with severe hypothermia and respiratory insufficiency after submersion in water. The ECG showed Osborn waves and a prolonged QRS and QT duration which normalized after correction of the hypothermia. PMID- 15524132 TI - [Body-mass index cut-off values for underweight in Dutch children]. AB - OBJECTIVE: To determine the cut-off values for the body-mass index (BMI) for underweight and serious underweight in children up to 18 years of age based on the Dutch growth standards of 1980, and to determine the prevalence of underweight and serious underweight in the 1997 Dutch growth-study figures. DESIGN: Secondary data analysis. METHOD: Cut-off values for underweight were determined by first calculating the percentage of 18-years old with a BMI < 18.5 kg/m2 in 1980, followed by a calculation of the BMI at younger ages under which the same percentage fell. Serious underweight in 18-year olds was defined as a BMI < 17.0 kg/m2. RESULTS: Between 1980 and 1997 the prevalence of serious underweight increased in children aged between 2 and 6 years from 1.5% to 3.6% and from 2.9% to 3.3%, in boys and girls respectively. In older children (7-18 years) the prevalence decreased from 1.5% to 1.4% for boys and from 2.9% to 1.7% for girls. CONCLUSION: Not only did the median weight of the population increase between 1980 and 1997, but there was also a rise in the variation of the weights at any given age. The latter could reflect more marked health differences. PMID- 15524133 TI - [Activity loss of Von Willebrand factor cleaving protein (ADAMTS-13) is diagnostic for primary and pregnancy-related thrombotic thrombocytopenic purpura]. AB - OBJECTIVE: To determine whether the measurement of the Von Willebrand factor cleaving protease ADAMTS-13, such as is carried out at the University Medical Centre of Utrecht, The Netherlands, contributes towards the diagnosis and treatment of patients with thrombotic thrombocytopenic purpura (TTP). DESIGN: Descriptive. METHOD: In a group of 98 patients from 21 hospitals, with a Coombs negative haemolytic anaemia and thrombocytopenia, the ADAMTS-13 activity was measured. Treatment was given irrespective of ADAMTS-13 activity. RESULTS: ADAMTS 13 activity was absent in 27 of 29 patients diagnosed with primary TTP and in all 5 pregnancy-TTP patients. In patients suffering from TTP after bone marrow transplantation (post-BMT) and in all other patients included in this study, ADAMTS-13 activity was normal. Of the 32 patients with absent ADAMTS-13 activity, 28 underwent plasmapheresis. This treatment proved effective as all 28 patients responded well. 17 patients with normal ADAMTS-13 activity also underwent plasmapheresis; 5 (30%) responded well to treatment. In 2 cases a final diagnosis of primary TTP was made, in a further 2, haemolytic uraemic syndrome and in 1 case sepsis was diagnosed. CONCLUSION: In this study, the absence of ADAMTS-13 activity predicted primary TTP and TTP of pregnancy with a sensitivity of 93% and a specificity of 100%. Absence of ADAMTS-13 activity is a strong indication for plasma exchange. PMID- 15524134 TI - [A woman suffering from systemic lupus erythematosus and acquired thrombotic thrombocytopenic purpura]. AB - A 41-year-old woman with known systemic lupus erythematosus (SLE) was diagnosed with thrombotic thrombocytopaenic purpura (TTP). At the time of admission she was suffering from petechia, purpura and had neurological symptoms. At first a relapse of the SLE was suspected. Additional laboratory findings demonstrated haemolytic anaemia, thrombocytopaenia and high levels of fragmentocytes. After multiple plasmapheresis treatments and immunosuppressive therapy she recovered. TTP can be differentiated from other thrombotic microangiopathic syndromes by its normal levels of prothrombin time, partially activated thromboplastin time (APTT), fibrinogen and direct Coombs-test. Further investigation is needed to confirm the diagnosis by determination of the activity of Von Willebrand factor cleaving protease ADAMTS-13. In this patient, the ADAMTS-13 activity returned after plasmapheresis. This case demonstrates the importance of fast and appropriate laboratory testing in order to diagnose TTP quickly. PMID- 15524135 TI - [Cryopreservation of semen of adolescents and young adult men with cancer]. AB - The importance of cryopreserving semen for young male cancer patients is illustrated in three case descriptions. A 28-year-old man with chronic myeloid leukaemia that resulted in azoospermia, later fathered a child with his semen that had been stored prior to chemotherapy. In an 18-year-old adolescent with non Hodgkin lymphoma the possibility to store cryopreserved semen was only raised after chemotherapy had been started and had caused azoospermia. This caused the patient serious regret. A 14-year-old boy with acute lymphatic leukaemia had his semen stored despite initial hesitations due to his young age. The cancer hardly ever affects the semen quality to the extent that cryopreservation of the semen becomes impossible. The aim should be to obtain several ejaculates prior to the cancer therapy and to store multiple portions, so that later a number of fertilisation attempts are possible. The primary attending physician is initially responsible for raising the possibility of semen cryopreservation. Ideally, however, all health professionals involved should be aware of this aspect. There is a need for multidisciplinary protocols for oncology centres and sperm banks, so that the timely informing of patients is guaranteed, responsibilities are recorded--with appropriate procedures to prevent unnecessary delay--and procedures that concur with legal requirements and financial constraints are established. PMID- 15524137 TI - [Manipulation of mental competence: an ethical problem in case of electrical stimulation of the subthalamic nucleus for severe Parkinson's disease]. PMID- 15524136 TI - [The 2003/2004 influenza season in the Netherlands with a limited epidemic of the virus variant A/Fujian, and the vaccine composition for the 2004/2005 season]. AB - In contrast to the three previous influenza seasons, the influenza epidemic of the 2003/2004 season started early in week 49 of 2003. The epidemic was predominantly caused by influenza-A viruses of the H3N2 subtype. All isolated influenza-A viruses were antigenically related to influenza virus A/Fujian/411/02, which was already detected in the influenza season 2002/2003 and that deviated from the vaccine-reference strain A/Moscow/10/99 to a certain extent. The magnitude of the epidemic was limited despite the fact that it was caused by influenza-A H3N2-virus-drift variants. Immunity caused by natural infection with influenza viruses during previous seasons or vaccination has possibly provided sufficient cross protection against these new H3N2-drift variant. No influenza-A viruses of the H1N1 or H1N2 subtypes were detected in the influenza season 2003/2004. Only a small number of influenza-B viruses were isolated, which all belonged to the B/Yamagata/16/88 lineage, which was temporarily replaced by the B/Victoria2/87 lineage in the previous influenza season. On the basis of epidemiological and serological data the World Health Organization has recommended the following vaccine composition for the 2004/2005 influenza season: A/Fujian/411/02 (H3N2), A/New Caledonia/20/99 (H1N1) and B/Shanghai/361/02. PMID- 15524138 TI - [Manipulation of mental competence: an ethical problem in case of electrical stimulation of the subthalamic nucleus for severe Parkinson's disease]. PMID- 15524139 TI - [Invasive prenatal diagnosis in the Netherlands, 1991-2000: number of procedures, indications and abnormal results detected]. PMID- 15524140 TI - [Summary of the practice guideline 'Hormonal contraception' (second revision) from the Dutch College of General Practitioners]. PMID- 15524141 TI - Protecting the home front: experts recommend practical steps to help you plan for the unthinkable. PMID- 15524142 TI - A model of preparedness: State associations partner for disaster response. PMID- 15524143 TI - Syndromic surveillance, disaster preparedness & home care: is there a connection? PMID- 15524144 TI - Revised CDC isolation guideline takes home care into account. PMID- 15524145 TI - Robert M. Ball: a tribute to a gentleman and a scholar. PMID- 15524146 TI - Getting serious about the politics of home care: the Texas approach. PMID- 15524147 TI - President develops health information technology plan. PMID- 15524148 TI - CMS responds to further queries about medicare hospice benefit. PMID- 15524149 TI - Question thyself. PMID- 15524150 TI - The world of private duty home care. PMID- 15524151 TI - Respecting individual belief systems. PMID- 15524152 TI - "Granny Annie". PMID- 15524153 TI - Shoring up congressional support for post-election year politics. PMID- 15524154 TI - [Screening for Y chromosome sequences in patients with Turner syndrome]. AB - The Turner syndrome (TS) has been described in association with different sex chromosome aberrations. Although most TS patients show no evidence of Y chromosome sequences, according to different authors some TS patients may have Y chromosome material present in a few cells that are not detected by standard cytogenetic analysis. The importance of identification of this low level Y mosaicism is of clinical relevance due to the patient's increased risk of developing gonadoblastoma. In the present study, standard chromosome analysis performed on peripheral blood lymphocytes from 22 TS patients showed 12 patients with 45,X karyotype, 7 patients were mosaics with or without structural abnormalities in one X chromosome and, the remaining three patients had the following karyotypes: 46,X,i(X)(q10); 46,X,+mar/47,X,idic(Y),+mar and 45,X/46,X,+r. Molecular studies were performed on genomic DNA extracted from peripheral blood lymphocytes and mouth epithelial cells, which derive from two different embryonic germ layers, mesoderm and ectoderm, respectively. The screening for low level Y mosaicism was carried out by simplex PCR and by nested PCR of the following Y specific loci: SRY (sex determining region Y), TSPY (testis specific protein Y encoded), DYZ3 (centromeric locus) and DAZ1 (deleted in azoospermia). In two TS patients a set of STSs of the Y long and short arms were used to characterize the idic(Y) and the ring chromosomes. The high sensitivity of the nested PCR (1 male cell/125,000 female cells) allowed for exclusion of the presence of low level Y mosaicism in 20 out of 22 TS patients. In the patient with the idic(Y), PCR analysis was positive for all Y loci tested excluding the heterochromatic region. This result identified the breakpoint between sY158 and sY159 on the long arm and, by fluorescence in situ hybridization (FISH) it was confirmed that the euchromatic part of the long arm, centromere and short arm of the Y chromosome were duplicated. The characterization of the ring chromosome, detected in one of the TS patients, was only possible to analyse by FISH and PCR. In this ring, derived from the Y chromosome, a deletion was identified including the pseudoautosomal region 1 (PARY1) and Y intervals 6 and 7. However, the ring Y was positive for SRY, RPS4Y, AMGY, TSPY loci on the short arm, DYZ3 (centromere) and, sY85, DFFRY, GY6, sY87, sY113, sY119, sY122, sY126 and RBMY1 on the long arm. This study excluded the presence of low level Y mosaicism in two tissues collected from 20 TS patients. FISH and molecular analysis allowed us to characterize, in 2 out of 22 patients, one idic(Y) and one ring chromosome. The nature of the latter had not been completely identified by standard cytogenetics. The potential increased risk of gonadoblastoma in TS patients carrying Y chromosome sequences justifies the application of FISH and PCR for the characterization of marker chromosomes and the application of nested PCR for the detection of low level Y mosaicisms when Y chromosome material is not detected by standard cytogenetic analysis in patients with a 45,X karyotype and/or with virilization. PMID- 15524155 TI - [Endemic goiter epidemiological study of prevalence in Oleiros]. AB - This research was carried out in the county of Oleiros, which integrates the only defined endemic goiter zone in Portugal, since 1969. We intend to characterize, from the epidemiological point of view, the prevalence of the endemic goiter by the observation of all students (N=699) and by the determination of the iodine urinary excretion in a sample of them (n=43). The prevalence value was of 27.5% (192 in 699), corresponding to 21.6% and to 5.9%, respectively for the total cases of degree 1A and degrees 1B and 2. No cases of degree 3 were found. Prevalence was of 36.4% (116 in 319) for females, and 20% (76 in 380) for males (proportion of about 2:1). The iodine urinary excretions presented 20% of the values out of the normal limits, and about 10% of them below minimum. These results allow us to classify the region as one of low to moderate endemic severity. However, in order to eradicate, among us, the endemic goiter, it is essential to proceed with the prophylactic program. PMID- 15524156 TI - [Assessement of an automated homogeneous liposome-based immunoassay system for total complement activity]. AB - The Immunology department of Garcia de Orta Hospital, has evaluated a new automated homogeneous liposome-based immunoassay (L.I.A.), for measuring total complement activity, which has become recently commercially available from WAKO (Osaka, Japan), (CH50 WAKO). Serum samples from 100 healthy voluntary donors were assayed, with liposome-based assay. The reference values in our adult population concerning CH50 were established. Within-run and between-run precision, accuracy, carry-over and the linearity of the new method were also assessed. PMID- 15524157 TI - [Craniopharyngiomas. Clinicopathological aspects in different age groups]. AB - Craniopharyngiomas are rare brain tumors of the hypothalamo-pituitary region, developing from embryonic remnants of Rathke's pouch and sac. Their overall incidence is 0.13 per 100,000 person years. Most frequently, they are suprasellar, start growing in childhood and originate neurological and hormonal symptoms. We retrospectively studied patients treated in our institution for craniopharyngioma in the last 10 years, in order to evaluate their clinical, imaging and pathological characteristics. Of the 32 patients analysed, 18 were females and 14 males with ages ranging between 6 and 81 years (early onset group- EOG aged 5-14 years: 7 patients; middle age onset group--MAOG aged 15-49 years: 15 patients; late age onset group--LOG aged > or = 50 years: 10 patients). Visual impairment was the most frequent presenting clinical feature in EOG (71.4%) and MAOG (86.6%), while in the LOG personality and cognitive changes including memory loss predominated (60%). Headaches were very frequent in all groups (EOG 42.8%, MAOG 60%, LOG 40%). Meningitis and seizures were presenting features, each in one patient. Regarding endocrine symptoms and signs, growth failure was present in 57.2% of the EOG. Amenorrhea was present in 5 of 10 female patients of the MAOG. Preoperatively, TSH was deficient in 25%, ACTH in 15.6% and gonadotropin in 25% of the patients. There were no cases of diabetes insipidus. Preoperative CT and MR revealed a calcified mass in 12 (37.5%), a partially cystic mass in 20 (62.5%) and a lesion involving or extending into the third ventricle in 7 (21.9%) patients. Twenty seven (84.4%) patients were treated primarily by surgery. In 4 (12.5%) cases the tumour was considered inoperable and 1 (3.1%) patient refused surgery; all were in the LOG. Surgical approach was transsphenoidal in 2/27 (7.4%) (all of them in the LAOG) and by craniotomy in the others. The tumour removable was considered complete in 10 (37%--EOG 2/7, MAOG 6/15, LOG 2/5) and subtotal in 17 (62.9%) patients. Eight (29.6%) patients were reoperated for recurrent tumour. Postoperative radiotherapy was administered in 12 cases with residual tumor, and 3 inoperable tumors were treated primarily by conventional external radiotherapy. Pathological study revealed the adamantinomatous type in 25 (92.6%) and the papillary type in 2 (7.4%--all men in the MAOG) tumors. The average follow-up was longer in the EOG (82.6 +/- 40.7 months) than in MAOG (57.2 +/- 48.5 months) and in LOG (48 +/- 92 months). Four (12.5%) patients died, 1 during the follow-up period due to a radiation-induced astrocytoma and 3 in the postoperative period because of cerebral hemorrhage and hydrocephalus (1 in the EOG and 2 in the LOG). In summary, we found the clinical presentation to be different in the 3 age groups, with a large number of patients in the MAOG. In this group were the only examples of the papillary form. Better prognosis was associated with a total resection at initial surgery. PMID- 15524158 TI - [DNA flow cytrometry in solid tumors]. AB - This brief overview outlines the fundamental principles of flow cytometry with emphasis on DNA measurements and cell cycle analysis in human solid tumors. Type of material used, sampling processing procedures and methods of analysis of data are discussed. DNA ploidy and proliferative activity (S-phase fraction) are the two biological parameters commonly measured by DNA flow cytometric analysis. The prime purpose of most studies in this area is the investigation of the prognostic value of DNA flow cytometry in addition to the information provided by conventional clinicopathological factors known to affect disease prognosis. Numerous studies concerning the predictive significance of DNA flow cytometry in some types of solid tumors are reviewed in this article. The general statement, for tumors in the same histopathological stage of the disease, is that diploid and/or low proliferative tumors have a more favourable prognosis than aneuploid and/or high proliferative tumors, suggesting an important role of DNA flow cytometry in the assessment of tumor behaviour and in the outcome evaluation of the disease. However, in some studies this association could not be substantiated, and the prognostic relevance of DNA analysis has been questioned. The potential reasons for conflicting results, namely the methodological pitfalls related to the cell preparation techniques and the histogram interpretation are discussed. PMID- 15524159 TI - [Aircraft cabin air quality: exposure to ozone]. AB - Ozone is the principal component involved in photochemical pollution of the air. As an irritant of the respiratory system, its effects on the health of those exposed to it are characterised essentially by coughing, shortness of breath, chest pain or tightness and alterations to the pulmonary mechanical function. Additionally, a higher frequency and severity of asthmatic exacerbation and the occurrence of eye irritation are linked to environmental exposure to O3. In the early 1960s the first studies on the exposure to O3 in aircraft cabins appeared, prompted by the occurrence of clinical complaints of irritation of the respiratory tract in crewmembers and passengers. The symptoms had hitherto been attributed to the action of other factors, such as the ventilation system and low level of humidity in the air. An updating is done by author of some factors related to the quality of air inside aircraft cabins, namely the exposure to ozone in crewmembers and passengers. PMID- 15524160 TI - [Acute disautonomia associated to Hodgkin lymphoma]. AB - The case of a 31 man with acute disautonomia envolving the parasympathetic and sympathetic systems but sparing sympathetic cholinergic division is presented. A Hodgkin lymphoma was diagnosed allowing the diagnosis of a paraneoplastic syndrome. PMID- 15524161 TI - [Cardiovascular disease and pregnancy]. AB - Cardiovascular disease is the most important non-obstetric cause of disability and death in pregnant women, occurring in 1% of pregnancies. The added hemodynamic burden of pregnancy, labor and delivery can aggravate symptoms and precipitate complications in women with pre-existent cardiac disease. Additionally, it can reveal an underlying unknown pathology. The authors present three cases of pregnant women with mechanical prosthetic valve, cyanotic congenital heart disease and intracardiac shunt. Antenatal, labor and postpartum obstetric management is subject of debate. PMID- 15524162 TI - Genetic and phenotypic targeting of beta-adrenergic signaling in heart failure. AB - Heart failure is a leading cause of hospitalization worldwide. No major significant improvements in prognosis have been achieved for heart failure over the last several decades despite advances in disease management. Heart failure itself represents a final common endpoint for several disease entities, including hypertension and coronary artery disease. On a molecular level, certain biochemical features remain common to failing myocardium. Among these are alterations in the beta-adrenergic receptor (beta-AR) signaling cascade. Recent advances in transgenic and gene therapy techniques have presented novel therapeutic strategies for management of heart failure via genetic manipulation of beta-AR signaling including the targeted inhibition of the beta-AR kinase (betaARK1 or GRK2). In this review, we will discuss the beta-AR signaling changes that accompany heart failure as well as corresponding therapeutic strategies. We will then review the evidence from transgenic mouse work supporting the use of beta-AR manipulation in the failing heart and more recent in vivo applications of gene therapy directed at reversing or preventing heart failure. PMID- 15524163 TI - Changes in beta-adrenoceptors in heart failure due to myocardial infarction are attenuated by blockade of renin-angiotensin system. AB - Earlier studies have revealed an improvement of cardiac function in animals with congestive heart failure (CHF) due to myocardial infarction (MI) by treatment with angiotensin converting enzyme (ACE) inhibitors. Since heart failure is also associated with attenuated responses to catecholamines, we examined the effects of imidapril, an ACE inhibitor, on the beta-adrenoceptor (beta-AR) signal transduction in the failing heart. Heart failure in rats was induced by occluding the coronary artery, and 3 weeks later the animals were treated with g/(kg x day) (orally) imidapril for 4 weeks. The animals were assessed for their left ventricular function and inotropic responses to isoproterenol. Cardiomyocytes and crude membranes were isolated from the non-ischemic viable left ventricle and examined for the intracellular concentration of Ca2+ [Ca2+]i and beta-ARs as well as adenylyl cyclase (AC) activity, respectively. Animals with heart failure exhibited depressions in ventricular function and positive inotropic response to isoproterenol as well as isoproterenol-induced increase in [Ca2+]i in cardiomyocytes; these changes were attenuated by imidapril treatment. Both beta1 AR receptor density and isoproterenol-stimulated AC activity were decreased in the failing heart and these alterations were prevented by imidapril treatment. Alterations in cardiac function, positive inotropic effect of isoproterenol, beta1-AR density and isoproterenol-stimulated AC activity in the failing heart were also attenuated by treatment with another ACE inhibitor, enalapril and an angiotensin II receptor antagonist, losartan. The results indicate that imidapril not only attenuates cardiac dysfunction but also prevents changes in beta-AR signal transduction in CHF due to MI. These beneficial effects are similar to those of enalapril or losartan and thus appear to be due to blockade of the renin angiotensin system. PMID- 15524164 TI - The endothelium in health and disease: a discussion of the contribution of non nitric oxide endothelium-derived vasoactive mediators to vascular homeostasis in normal vessels and in type II diabetes. AB - Endothelial dysfunction is considered as a major risk factor of cardiovascular complications of type I and types II diabetes. Impaired endothelium-dependent vasodilatation can be directly linked to a decreased synthesis of the endothelium derived nitric oxide (NO) and/or an increase in the production of reactive oxygen species such as superoxide. Administration of tetrahydrobiopterin, an important co-factor for the enzyme nitric oxide synthase (NOS), has been demonstrated to enhance NO production in prehypertensive rats, restore endothelium-dependent vasodilatation in coronary arteries following reperfusion injury, aortae from streptozotocin-induced diabetic rats and in patients with hypercholesterolemia. Tetrahydrobiopterin supplementation has been shown to improve endothelium dependent relaxation in normal individuals, patients with type II diabetes and in smokers. These findings from different animal models as well as in clinical trials lead to the hypothesis that tetrahydrobiopterin, or a precursor thereof, could be a new and an effective therapeutic approach for the improvement of endothelium function in pathophysiological conditions. In addition to NO, the endothelium also produces a variety of other vasoactive factors and a key question is: Is there also a link to changes in the synthesis/action of these other endothelium-derived factors to the cardiovascular complications associated with diabetes? Endothelium-derived hyperpolarizing factor, or EDHF, is thought to be an extremely important vasodilator substance notably in the resistance vasculature. Unfortunately, the nature and, indeed, the very existence of EDHF remains obscure. Potentially there are multiple EDHFs demonstrating vessel selectivity in their actions. However, until now, identity and properties of EDHF that determine the therapeutic potential of manipulating EDHF remains unknown. Here we briefly review the current status of EDHF and the link between EDHF and endothelial dysfunction associated with diabetes. PMID- 15524165 TI - The effect of nitric oxide on glucose metabolism. AB - Nitric oxide (NO) is an important bioactive signaling molecule that mediates a variety of normal physiological functions, which, if altered, could contribute to the genesis of many pathological conditions, including diabetes. In this study, we examined the possible diabetogenicity of NO by noting differences in the cellular binding of insulin in dogs treated with the NO donor, S nitrosoglutathione (GSNO) compared to captopril-treated controls. GSNO administration resulted in an abnormality in glucose metabolism which was attributed to decreased binding of insulin to its receptor on the cell membrane of mononuclear leucocytes, 11.60 +/- 0.60% in GSNO-treated dogs compared with 18.10 +/- 1.90% in captopril-treated control (p < 0.05). The decreased insulin binding was attributed to decreased insulin receptor sites per cell, 21.43 +/- 2.51 x 10(4) in GSNO-treated dogs compared with 26.60 +/- 1.57 x 10(4) in captopril-treated controls (p < 0.05). Average affinity analysis of the binding data demonstrated that this decrease in insulin binding was also due to a decrease in average affinity of the receptor on mononuclear leucocytes for insulin. This was evident by a decrease in empty and filled site affinities in GSNO-treated dogs compared with that of captopril-treated dogs (p < 0.05). It appears that GSNO is exerting its effect by decreasing the number of insulin receptor sites and/or decreasing the average receptor affinity. These results provide evidence for a novel role of NO as a modulator of insulin binding and the involvement of NO in the aetiology of diabetes mellitus. PMID- 15524166 TI - Nitric oxide and promotion of cardiac myocyte apoptosis. AB - The removal of damaged, superfluous or energy-starved cells is essential for biological homeostasis, and occurs in every tissue type. Programmed cell death occurs through several closely regulated signal pathways, including apoptosis, in which cell components are broken down and packaged into small membrane-bound fragments that are then removed by neighbouring cells or phagocytes. This process is activated in the cardiac myocyte in response to a variety of stresses, including oxidative and nitrosative stress, and involves mitochondria-derived signals. Loss of cardiac myocytes through apoptosis has been shown to induce cardiomyopathy in a variety of gene-targeted animal models. Because cardiac myocytes have strictly limited ability to regenerate, sustained programmed cell death is likely to contribute to the development and progression of heart failure in a variety of myocardial diseases. At the same time, the cardiac myocyte possesses a number of mechanisms for defence against short-term haemodynamic and oxidative stresses. Our laboratory has recently examined the role of nitric oxide (NO) as a regulator of the programmed death of cardiac myocytes, and the potential contribution of NO and NO-dependent signalling to the loss of myocytes in heart failure. We will review the role of c-Jun N-terminal kinase in response to oxidative and nitrosative stress, and summarise evidence for its role as a cytoprotective mechanism. We will also review evidence implicating NO in the pathophysiology of heart failure, in the context of the extensive and sometimes contradictory body of research on NO and cell survival. PMID- 15524167 TI - Autophagy: many paths to the same end. AB - Different mechanisms lead to the degradation of intracellular proteins in the lysosomal compartment. Activation of one autophagic pathway or another, under specific cellular conditions, plays an important role in the ability of the cell to adapt to environmental changes. Each form of autophagy has its own individual characteristics, but it also shares common steps and components with the others. This interdependence of the autophagic pathways confers to the lysosomal system, both specificity and flexibility on substrate degradation. We describe in this review some of the recent findings on the molecular basis and regulation for each of the different autophagic pathways. We also discuss the cellular consequences of their interdependent function. Malfunctioning of the autophagic systems has dramatic consequences, especially in non-dividing differentiated cells. Using the heart as an example of such cells, we analyze the relevance of autophagy in aging and cell death, as well as in different pathological conditions. PMID- 15524168 TI - Reduced cross-bridge dependent stiffness of skinned myocardium from mice lacking cardiac myosin binding protein-C. AB - The role of cardiac myosin binding protein-C (MyBP-C) on myocardial stiffness was examined in skinned papillary muscles of wild-type (WT(+/+)) and homozygous truncated cardiac MyBP-C (MyBP-C(t/t) male mice. No MyBP-C was detected by gel electrophoresis or by Western blots in the MyBP-C(t/t) myocardium. Rigor-bridge dependent myofilament stiffness, i.e., rigor minus relaxed stiffness, in the MyBP C(t/t) myocardium (281 +/- 44 kN/m2) was 44% that in WT(+/+) (633 +/- 141 kN/m2). The center-to-center spacing between thick filaments as determined by X-ray diffraction in MyBP-C(t/t) (45.0 +/- 1.2 nm) was not significantly different from that in WT(+/+) (43.2 +/- 0.9 nm). The fraction of cross-sectional area comprised of myofibrils, as determined by electron microscopy, was reduced in the MyBP C(t/t) (39.9%) by 10% compared to WT(+/+) (44.5%). These data suggest that the 56% reduction in rigor-bridge dependent stiffness of the skinned MyBP-C(t/t) myocardium could not be due solely to a 10% reduction in the number of thick filaments per cross-sectional area and must also be due to approximately 50% reduction in the stiffness of the rigor-bridge attached thick filaments lacking MyBP-C. PMID- 15524169 TI - Structure and regulation of human troponin genes. AB - The recent completion of a first draft of the human genome has allowed "in silico" genome browsing to become routine. Such computer-based research is now a useful adjunct to experiments based at the bench, and is accelerating gene discovery and the analysis and understanding of genes in their genomic contexts. This review summarises recent findings on genes encoding proteins of the troponin complex. We describe the organization of the three pairs of genes which encode isoforms of troponins I and T, and discuss how this relates to their evolution and regulation. Detailed analysis of the chromosomal context of the cardiac troponin I and slow skeletal troponin T genes reveals a region of densely packed differentially expressed genes, including new genes identified by automatic genome annotation. This information is discussed within the context of detailed analysis of the best-studied gene in this region, cardiac troponin I. In this way, we illustrate the uses to which a combination of conventional bench experiments and "in silico" analyses may be put in understanding the relationship between structure and function within the genome. PMID- 15524170 TI - The slow skeletal muscle troponin T gene is expressed in developing and diseased human heart. AB - Cardiac muscle development is characterised by the activation of contractile protein genes and subsequent modulation of expression resulting, ultimately, in the formation of a mature four-chambered organ. Myocardial gene expression is also altered in the adult in response to pathological stimuli and this is thought to contribute to the altered contractile characteristics of the diseased heart. We have examined the expression of the slow skeletal troponin T (TnT) gene in the human heart during development and in disease using whole mount in situ hybridisation and real-time quantitative (TaqMan) polymerase chain reaction (PCR). Slow skeletal TnT mRNA shows transitory and regional expression in the early foetal heart, which occurs at different times in atria and ventricles. In ventricular myocardium, expression is seen in the outer epicardial layer at a time when the coronary circulation is being established. Expression was detected at low levels in the adult human heart and was significantly increased in end stage heart failure. Similarly, expression was readily detectable during early rat heart development and was up-regulated in pressure overload hypertrophy in adult. Together these data show for the first time that slow skeletal TnT mRNA is readily detectable during early human heart development. They further suggest that slow skeletal TnT may be responsive to myocardial stress and that elevated levels may contribute to myocardial dysfunction in adult disease. PMID- 15524171 TI - Cellular and molecular aspects of familial hypertrophic cardiomyopathy caused by mutations in the cardiac troponin I gene. AB - Mutations in the cardiac troponin I (CTnI) gene occur in approximately 5% of families with familial hypertrophic cardiomyopathy (FHC) and 20 mutations in this gene that cause FHC have now been described. The clinical manifestations of CTnI mutations that cause FHC are diverse, ranging from asymptomatic with high life expectancy to severe heart failure and sudden cardiac death. Most of these FHC mutations in CTnI result in cardiac hypertrophy unlike cardiac troponin T FHC mutations. All CTnI FHC mutations investigated in vitro affect the physiological function of CTnI, but other factors such as environmental or genetic factors (other genes that may affect the CTnI gene) are likely to be involved in influencing the severity of the phenotype produced by these mutations, since the distribution of hypertrophy among affected individuals varies within and between families. CTnI mutations mainly alter myocardial performance via changes in the Ca2+ -sensitivity of force development and in some cases alter the muscle relaxation kinetics due to haemodynamic or physical obstructions of blood flow from the left ventricle. PMID- 15524172 TI - Role of troponin T in disease. AB - Several striated muscle myopathies have been directly linked to mutations in contractile and associated proteins. Troponin T (TnT) is one of the three subunits that form troponin (Tn) which together with tropomyosin is responsible for the regulation of striated muscle contraction. All three subunits of cardiac Tn as well as tropomyosin have been associated with hypertrophic cardiomyopathy (HCM). However, TnT accounts for most of the mutations that cause HCM in these regulatory proteins. To date 30 mutations have been identified in the cardiac TnT (CTnT) gene that results in familial HCM (FHC). The CTnT gene has also been associated with familial dilated cardiomyopathy (DCM). CTnT deficiency is lethal due to impaired cardiac development. A recessive nonsense mutation in the gene encoding slow skeletal TnT has been associated with an unusual, severe form of nemaline myopathy among the Old Order Amish. How each mutation leads to the diverse clinical symptoms associated with FHC, DCM or nemaline myopathy is unclear. However, the use of animal model systems, in particular transgenic mice, has significantly increased our knowledge of normal and myopathic muscle physiology. In this review, we focus on the role of TnT in muscle physiology and disease. PMID- 15524173 TI - Phosphorylation of phospholamban in ischemia-reperfusion injury: functional role of Thr17 residue. AB - Phospholamban (PLB) is a sarcoplasmic reticulum (SR) protein that when phosphorylated at Ser16 by PKA and/or at Thr17 by CaMKII increases the affinity of the SR Ca2+ pump for Ca2+. PLB is therefore, a critical regulator of SR function, myocardial relaxation and myocardial contractility. The present study was undertaken to examine the status of PLB phosphorylation after ischemia and reperfusion and to provide evidence about the possible role of the phosphorylation of Thr17 PLB residue on the recovery of contractility and relaxation after a period of ischemia. Experiments were performed in Langendorff perfused hearts from Wistar rats. Hearts were submitted to a protocol of global normothermic ischemia and reperfusion. The results showed that (1) the phosphorylation of Ser16 and Thr17 residues of PLB increased at the end of the ischemia and the onset of reperfusion, respectively. The increase in Thr17 phosphorylation was associated with a recovery of relaxation to preischemic values. This recovery occurred in spite of the fact that contractility was depressed. (2) The reperfusion-induced increase in Thr17 phosphorylation was dependent on Ca2+ entry to the cardiac cell. This Ca2+ influx would mainly occur by the coupled activation of the Na+ / H+ exchanger and the Na+ / Ca2+ exchanger working in the reverse mode, since phosphorylation of Thr17 was decreased by inhibition of these exchangers and not affected by blockade of the L-type Ca2+ channels. (3) Specific inhibition of CaMKII by KN93 significantly decreased Thr17 phosphorylation. This decrease was associated with an impairment of myocardial relaxation. The present study suggests that the phosphorylation of Thr17 of PLB upon reflow, may favor the full recovery of relaxation after ischemia. PMID- 15524174 TI - Calreticulin in the heart. AB - Calreticulin is a Ca2+ binding/storage chaperone resident protein of the endoplasmic reticulum. This protein plays a key role in the calreticulin/calnexin cycle and the quality control pathways in the endoplasmic reticulum. Calreticulin deficiency is lethal due to impaired cardiac development. However, over expression of the protein in developing and postnatal heart leads to bradycardia, complete heart block and sudden death. Ultrastructural evidence indicates that the deficiency associated with the absence of calreticulin in the heart may be due to a defect in the development of the contractile apparatus and/or a defect in development of the conductive system as well as a metabolic abnormality. Collectively, we postulate that calreticulin and endoplasmic reticulum plays an important role in cardiac development and postnatal pathologies. PMID- 15524176 TI - Mechanisms of low Na+-induced increase in intracellular calcium in KCl depolarized rat cardiomyocytes. AB - Although low Na+ is known to increase the intracellular Ca2+ concentration ([Ca2+]i) in cardiac muscle, the exact mechanisms of low Na+ -induced increases in [Ca2+]i are not completely defined. To gain information in this regard, we examined the effects of low Na+ (35 mM) on freshly isolated cardiomyocytes from rat heart in the absence and presence of different interventions. The [Ca2+]i in cardiomyocytes was measured fluorometrically with Fura-2 AM. Following a 10 min incubation, the low Na+ -induced increase in [Ca2+], was only observed in cardiomyocytes depolarized with 30 mM KCl, but not in quiescent cardiomyocytes. In contrast, low Na+ did not alter the ATP-induced increase in [Ca2+]i in the cardiomyocytes. This increase in [Ca2+]i due to low Na+ and elevated KCl was dependent on the extracellular concentration of Ca2+ (0.25-2.0 mM). The L-type Ca2+ -channel blockers, verapamil and diltiazem, at low concentrations (1 microM) depressed the low Na+, KCl-induced increase in [Ca2+]i without significantly affecting the response to low Na+ alone. The low Na+, high KCl-induced increase in [Ca2+]i was attenuated by treatments of cardiomyocytes with high concentrations of both verapamil (5 and 10 microM), and diltiazem (5 and 10 microM) as well as with amiloride (5-20 microM), nickel (1.25-5.0 mM), cyclopiazonic acid (25 and 50 microM) and thapsigargin (10 and 20 microM). On the other hand, this response was augmented by ouabain (1 and 2 mM) and unaltered by 5-(N-methyl-N-isobutyl) amiloride (5 and 10 microM). These data suggest that in addition to the sarcolemmal Na+ - Ca2+ exchanger, both sarcolemmal Na+ - K+ ATPase, as well as the sarcoplasmic reticulum Ca2+ -pump play prominent roles in the low Na+ -induced increase in [Ca2+]i. PMID- 15524175 TI - Diabetes decreases mRNA levels of calcium-release channels in human atrial appendage. AB - Patients with chronic diabetes mellitus usually develop reductions in rate and force of cardiac contractions. Since calcium-release channels (ryanodine receptors (RyRs) and inositol 1,4,5-trisphosphate receptors (IP(3)Rs)) play integral roles in effecting these processes, we rationalize that alterations in their expression may underlie these defects. To test this hypothesis, right atrial appendages were obtained from diabetic (65.0 +/- 4.5 years) and nondiabetic (56.2 +/- 2.6 years) patients undergoing coronary arterial by-pass grafting and reverse transcription-polymerase chain reactions were used to compare steady state levels of mRNA encoding the three major isoforms of RyRs and IP(3)Rs. In this study we did not detect either RyR1 or RyR3 in human atrial appendage. When compared with nondiabetic patients, mRNA encoding RyR2 from diabetic patients decreased by 74.2 +/- 6.2% (p < 0.01). Diabetes also significantly decreased steady-state levels of mRNA encoding the IP(3)Rs in human atrial appendage. IP(3)R1 decreased by 24.2 +/- 4.6%, IP(3)R2 decreased by 63.0 +/- 4.6% and IP(3)R3 decreased by 55.5 +/- 6.5%. Since a reduction in steady state mRNA is usually indicative of a decrease in protein levels, these data suggest that the decrease in chronotropy and inotropy seen in chronic diabetic patients may be due in part to a decrease in expression of calcium-release channels. PMID- 15524177 TI - Mechanisms of action of pH-induced effects on vascular smooth muscle. AB - It is clear that pH has many effects on vascular smooth muscle and the overall action of pH on force will depend on the type of vascular smooth muscle in question and the combined effects on all the potential modulatory mechanisms. The major effects of pH on force appear to be mediated via modulation of [Ca]i rather than changes in the sensitivity of the contractile machinery to Ca2+. There are still numerous gaps in our understanding of the actions of pH and as more data become available, we will be able to better understand the major mechanisms involved. PMID- 15524178 TI - Human VEGF165-myoblasts produce concomitant angiogenesis/myogenesis in the regenerative heart. AB - Bioengineering the regenerative heart may provide a novel treatment for heart failure. On May 14, 2002, a 55-year-old man suffering from ischemic myocardial infarction received 25 injections carrying 465 million cGMP-produced pure myoblasts into his myocardium after coronary artery bypass grafting. As on August 28, 2002, his EKG was normal and showed no arrhythmia. His ejection fraction increased by 13%. He no longer experienced shortness of breath and angina as he did before the treatment. Three myogenesis mechanisms were elucidated with 17 human/porcine xenografts using cyclosporine as immunosuppressant. Some myoblasts developed to become cardiomyocytes. Others transferred their nuclei into host cardiomyocytes through natural cell fusion. As yet others formed skeletal myofibers with satellite cells. De novo production of contractile filaments augmented the heart contractility. Human myoblasts transduced with VEGF165 gene produced six times more capillaries in porcine myocardium than in placebo. Xenograft rejection was not observed for up to 20 weeks despite cyclosporine discontinuation at 6 weeks. Pros and cons of autografts vs. allografts are compared to guide future development of heart cell therapy. PMID- 15524179 TI - AT1 receptor blockade alters metabolic, functional and structural proteins after reperfused myocardial infarction: detection using proteomics. AB - Angiotensin II (AngII) type 1 receptor (AT1R) blockers (ARBs) limit left ventricular (LV) dysfunction and necrosis after reperfused myocardial infarction (RMI) and proteomics can detect changes in protein levels after injury. We applied proteomics to detect changes in levels of specific protein in the ischemic zone (IZ) and non-ischemic zone (NIZ) of dog hearts after in vivo RMI (90 min of anterior ischemia; 120 min of reperfusion) and treatment with intravenous vehicle (control) and the ARBs valsartan or irbesartan (10 mg/kg) over 30 min before RMI. We also assessed LV function, infarction and apoptosis. Both ARBs limited the RMI-induced LV dysfunction, infarct size and apoptosis. Proteomics detected differential expression of 5 randomly selected proteins in the IZ compared to the NIZ after RMI: decrease in a subunit of ATP synthase isoform precursor (consistent with increased conversion to a subunit under metabolic stress), M chain creatine kinase (consistent with cellular damage) and ventricular myosin light chain-1 (consistent with structural damage and decreased contractility); and increase in NAD+ -isocitrate dehydrogenase (ICDH) and alpha subunit and ATP synthase D chain (mitochondrial, consistent with metabolic dysfunction). Importantly, changes in NAD+ -ICDH and ATP synthase D chain were reversed by ARB therapy. Thus, proteomics can detect regional changes in metabolic, contractile, and structural proteins after RMI and several of these proteins are favorably modified by ARBs, suggesting that they may be novel therapeutic targets. PMID- 15524180 TI - PPAR-gamma receptor agonists--a review of their role in diabetic management in Trinidad and Tobago. AB - The PPAR-gamma receptor agonists, as a relatively new and perhaps still not very widely used class of antidiabetic agent in the Caribbean and particularly the Trinidadian context, possess pharmacologic properties that certainly have been shown to have impact on many of the inflammatory, metabolic, biochemical and structural macrovascular aberrations that occur in the type 2 diabetic. Activation of PPAR(gamma) nuclear receptors regulates the transcription of insulin-responsive genes involved in the control of glucose production, transport, and utilization. PPAR(gamma)-responsive genes also participate in the regulation of fatty acid metabolism, an important contributory pathogenic factor in this subset of patients. The unique mode of action of this class of therapeutic agent addresses a range of anomalies occurring at the cellular and sub-cellular level that are injurious to the diabetic. My aim in addressing the issue of the potential impact of PPAR-gamma receptor agonists on cardiovascular disease (CVD) morbidity and mortality in the diabetic, is first, to seek to enhance both an awareness of, and greater familiarity among our own physicians, with this class of drug, and secondly, to effect a timely review of the recent literature as it relates to the tremendous possibilities for the potential clinical gains that might accrue from their use, in so far as this may serve to ameliorate the ravages of the CVD disease that all too tragically attends the type 2 diabetic, and more specifically those with the insulin resistance syndrome. PMID- 15524181 TI - Differential antioxidant properties of red wine in water soluble and lipid soluble peroxyl radical generating systems. AB - Red wine and its components have been shown to possess cardioprotective and anti atherogenic effects. Additionally, red wine and many of its components like catechin, epicatechin, rutin, transresveratrol and quercetin possess antioxidant properties. Oxidized low density lipoprotein (LDL) is involved in the development of an atherosclerotic lesion. Red wine, therefore, may be anti-atherogenic because of its antioxidant effects on LDL modification. This study examined the antioxidant effects of catechin, epicatechin, rutin, transresveratrol, quercetin and Merlot wines on LDL oxidation. Merlot was chosen because although other red wines have been tested, limited information exists for this variety. Oxidation was carried out with AAPH (2,2'-Azo-bis(2-amidinopropane) dihydrochloride) and AMVN (2,2'-Azo-bis(2,4-dimethylvaleronitrile)), as water and lipid soluble peroxyl radical generating systems (FRGS), respectively. This allowed us to determine the lipophilic antioxidant characteristics of the wine and its components. Conjugated diene assays were used to measure LDL oxidation over 6 hrs. In an AAPH system, all polyphenolic compounds except transresveratrol displayed an antioxidant effect. LDL oxidation by AAPH was also inhibited by aliquots of Merlot wine. No antioxidant effects were observed in an AMVN environment except for a mild antioxidant effect by quercetin. Surprisingly, incubation of LDL with Merlot wine strongly protected against oxidation by AMVN. In summary, the five phenolic compounds displayed antioxidant effects in a water soluble free radical generating system, but only quercetin showed this in a lipid soluble one. However, red wine inhibited LDL oxidation by both water and lipid soluble free radical generating systems. Our data suggest, therefore, that red wines contain unidentified antioxidants that provide protection against LDL oxidation within a lipid soluble environment. PMID- 15524182 TI - Omega-3 fatty acids from fish oils and cardiovascular disease. AB - Fish and fish oils contain the omega-3 fatty acids known as eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA). Epidemiological studies have shown an inverse relation between the dietary consumption of fish containing EPA/DHA and mortality from coronary heart disease. These relationships have been substantiated from blood measures of omega-3 fatty acids including DHA as a physiological biomarker for omega-3 fatty acid status. Controlled intervention trials with fish oil supplements enriched in EPA/DHA have shown their potential to reduce mortality in post-myocardial infarction patients with a substantial reduction in the risk of sudden cardiac death. The cardioprotective effects of EPA/DHA are widespread, appear to act independently of blood cholesterol reduction, and are mediated by diverse mechanisms. Their overall effects include anti-arrhythmic, blood triglyceride-lowering, anti-thrombotic, anti-inflammatory, endothelial relaxation, plus others. Current dietary intakes of EPA/DHA in North America and elsewhere are well below those recommended by the American Heart Association for the management of patients with coronary heart disease. PMID- 15524183 TI - Protease activated receptors in cardiovascular function and disease. AB - Recent studies have shown that a novel class of protease activated receptors (PARs), which are composed of seven transmembrane G protein-coupled domains, are activated by serine proteases such as thrombin, trypsin and tryptase. Although four types (PAR 1, PAR 2, PAR 3 and PAR 4) of this class of receptors have been identified, their discrete physiological and pathological roles are still being unraveled. Extracellular proteolytic activation of PARs results in the cleavage of specific sites in the extracellular domain and formation of a new N-terminus which functions as a tethered ligand. The newly formed tethered ligand binds intramolecularly to an exposed site in the second transmembrane loop and triggers G-protein binding and intracellular signaling. Recent studies have shown that PAR 1, PAR-2 and PAR-4 have been involved in vascular development and a variety of other biological processes including apoptosis and remodeling. The use of animal model systems, mainly transgenic mice and synthetic tethered ligand domains, have contributed enormously to our knowledge of molecular signaling and the regulatory properties of various PARs in cardiomyocytes. This review focuses on the role of PARs in cardiovascular function and disease. PMID- 15524184 TI - Role of proteases in the pathophysiology of cardiac disease. AB - Cardiovascular disease is a major cause of death and thus a great deal of effort has been made in salvaging the diseased myocardium. Although various factors have been identified as possible causes of different cardiac diseases such as heart failure and ischemic heart disease, there is a real need to elucidate their role for the better understanding of the cardiac disease pathology and formulation of strategies for developing newer therapeutic interventions. In view of the intimate involvement of different types of proteases in maintaining cellular structure, the role of proteases in various cardiac diseases has become the focus of recent research. Proteases are present in the cytosol as well as are localized in a number of subcellular organelles in the cell. These are known to use extracellular matrix, cytoskeletal, sarcolemmal, sarcoplasmic reticular, mitochondrial and myofibrillar proteins as substrates. Work from different laboratories using a wide variety of techniques has shown that the activation of proteases causes alterations of a number of specific proteins leading to subcellular remodeling and cardiac dysfunction. Inhibition of protease action by different drugs and agents, therefore, has a clinical relevance and is expected to form a part of new treatment paradigm for improving heart function. This review examines the biochemistry and localization of some of the proteases in the cardiac tissue in addition to identification of the sites of action of some protease inhibitors. PMID- 15524185 TI - A lanthanide hybrid cluster as a selective optical chemosensor for phosphate containing anions in aqueous solution. AB - An europium-chelating [2Fe-2S] cluster was used to assemble an optical molecular chemosensor highly selective for phosphate-containing anions. Phosphate, pyrophosphate, AMP, ADP, ATP, DNA and RNA were well distinguished by UV/V is absorption or fluorescence studies. PMID- 15524186 TI - Supercritical fluid clean-up of environmental samples for the analysis of polycyclic aromatic hydrocarbons using time-of-flight secondary ion mass spectrometry. AB - A novel sample-pretreatment method for time-of-flight secondary ion mass spectrometry (TOF-SIMS) was developed using supercritical fluid extraction (SFE). In SFE, the extraction efficiency of a certain organic matter is controlled by the pressure and temperature of supercritical CO2. Two-step SFE (1st step at 10 Mpa, 40 degrees C; 2nd step at 30 MPa, 120 degrees C) was applied to diesel exhaust particles containing many kinds of n-alkanes and aromatic species. n Alkanes and polycyclic aromatic hydrocarbons (PAHs) were extracted in the 1st and 2nd steps, respectively. This selectivity was utilized for the sample preparation of TOF-SIMS analysis. Diesel exhaust particles after the 1st step of extraction were analyzed with TOF-SIMS, aiming at PAHs as analytical targets. The obtained spectrum was simplified, and mass peaks of individual PAHs were easily assigned, because unwanted compounds, like n-alkanes, were selectively removed by SFE. Furthermore, a simple calculation elucidated the outline of the spectrum. PMID- 15524187 TI - Determination of polycyclic aromatic hydrocarbons in water by solid-phase microextraction and liquid chromatography. AB - This study describes the determination of polycyclic aromatic hydrocarbons (PAHs) in water using high-performance liquid chromatography (HPLC) coupled with fluorescence detection (FLD). Because individual PAHs are generally present in water only at trace levels, a sensitive and accurate determination technique is essential. The separation and detection of five PAHs were run completely within 25 min by the HPLC/FLD system with an analytical C18 column, a fluorescence detection, and acetonitrile-water gradient elution. Calibration graphs were linear with very good correlation coefficients (r > 0.9998), and the detection limits were in the range of 2-6 ng/l for five PAHs. Solid phase microextraction (SPME) was performed for sample pretreatment prior to HPLC-FLD determination, and the governing parameters were investigated. Compared to conventional methods, SPME has high recovery, saves considerable time, and reduces solvents waste. The extraction efficiencies of five PAHs were above 88% and the extraction times were 35 min in one pretreatment procedure. One particular discovery is that 1.5 M sodium monochloroactate (ClCH2COONa) can improve the extraction yield of PAH compounds more than other inorganic salts. The SPME-HPLC-FLD technique provides a relatively simple, convenient, practical procedure, which was here successfully applied to determine five PAHs in water from authentic water samples. PMID- 15524188 TI - Characterization of metal binding properties of rhamnogalacturonan II from plant cell walls by size-exclusion HPLC/ICP-MS. AB - The binding properties of metal ions to a pectic polysaccharide, rhamnogalacturonan II (RG-II), from plant cell walls were analyzed by size exclusion HPLC/ICP-MS. The dimeric RG-II borate complex (dRG-II-B) from sugar beet and red wine contained 0.8 - 1.1 mol/mol of B, 0.8 - 1.1 mol/mol of Ca, 0.1 0.3 mol/mol of Sr, and 0.03 - 0.07 mol/mol of Ba. The treatment of dRG-II-B with Sr2+, Ba2+, Pb2+ or La3+ exchanged the originally present Ca, Sr and Ba. In contrast, monomeric RG-II (mRG-II), which contained approximately 0.1 mol/mol of Ca, formed complexes with La3+, Eu3+, and Lu3+ added to the solution, but did not do so with Ca2+, Sr2+, Ba2+ and Pb2+. The HPLC/ICP-MS and HPLC/RI (refractive index detector) analysis of the partially hydrolyzed mRG-II that was treated with La3+ indicated that the side chains and backbone of mRG-II together form a lanthanoid binding site. PMID- 15524189 TI - HPLC profile of captopril disulfide that undergoes reversible cis-trans conversion among three isomers. AB - Peculiarly shaped chromatograms of some compounds that consist of two reversible isomers have been reported. Those of a compound that consists of three reversible isomers are described here. Because disulfide of captopril has two cis-trans convertible bonds, it exists in three forms of cis-cis, cis-trans and trans-trans isomers. The disulfide has produced various HPLC-UV profiles under various conditions, such as three split peaks, three peaks with bridging plateaus and one broad peak. The effect of the column temperature and the eluent flow rate on the profile was investigated and explained by interconversion among three isomers. A profile analysis could provide some kinetic constants of this reaction. PMID- 15524190 TI - Cyclodextrin-aided determination of iodate and bromate in drinking water by microcolumn ion chromatography with precolumn enrichment. AB - A selective and simple method for the determination of iodate (IO3-) and bromate (BrO3-) by microcolumn ion chromatography (IC) is presented. In this study, IO3- and BrO3- were determined as IBr2- and tribromide (Br3-), respectively, via a postcolumn reaction with bromide (Br) under acidic conditions with the aid of alpha-cyclodextrin (alpha-CD) in microcolumn IC. IO3- and BrO3- were selectively detected by the present method at a wavelength of 253 or 265 nm. The present system achieved good selectivity for IO3- and BrO3- as well as good repeatability under suitable conditions. Precolumn enrichment improved the detection limit, and allowed the determination of BrO3- in bottled water as low as sub microg L(-1) level in microcolumn IC. PMID- 15524191 TI - Characterization of the microscopic surface structure of the octadecylsilica stationary phase using a molecular-dynamics simulation. AB - The influence of the mobile-phase composition and temperature on the surface structure of the octadecylsilica (ODS) stationary-phase was investigated by applying a molecular-dynamics (MD) simulation. The molecular model to which the MD simulation was applied consisted of three parts: an amorphous silica base, dimethyloctadecylsilyl ligands and mobile-phase solvents. More detailed information on the effect of the mobile-phase composition was obtained by constructing larger molecular models than those used in our previous study. The thickness of the hydrocarbon layer of the stationary phase could be estimated based on the distance between the carbon atom located at the terminal end of the ODS ligand and the silica gel surface. The structural information obtained by the calculation showed good consistency with the experimentally observed values. The gauche fraction in the ODS ligand conformation could be also estimated to obtain a more detailed ligand conformation for each molecular model. It was found that as the temperature increased, the ligand conformation collapsed more. This trend was the same as the experimentally observed trends obtained by NMR, FT-IR, and Raman spectroscopic techniques. PMID- 15524192 TI - Enantiomeric separation of basic drugs with partially filled serum albumin as chiral selector in capillary electrophoresis. AB - A reliable method is presented for the chiral separation of three basic drugs (mexiletine, chlorpheniramine and propranolol) with serum albumins (human and porcine, HSA and PSA) as chiral selectors by capillary electrophoresis in combination with the partial filling technique. Based on the systematic optimization of operation variables, the chiral separation of mexiletine, chlorpheniramine and propranolol was achieved in the pH 7.4 phosphate buffer by using HSA, PSA and PSA as selectors, respectively. The chiral recognition ability of HSA and PSA was compared. HSA and PSA show a different chiral recognition ability for each of the three drugs. In addition, the association constants between enantiomeric drugs and proteins were determined to be 2.00 and 3.80 x 10(2) M(-1) for mexiletine and HSA, 0.59 and 1.12 x 10(3) M(-1) for chlorpheniramine and PSA, and 0.87 and 1.42 x 10(3) M(-1) for propranolol and PSA. The method for the chiral separation and determination of association constants possesses the advantages of simple performance, effective avoiding of the interference of the UV detection from protein, and lowering of the reagent consumption. PMID- 15524193 TI - Determination of glyphosate in water samples by alternating-current oscillopolarographic titration. AB - This paper describes a novel, simple and sensitive method for the determination of glyphosate in water samples by alternating-current oscillopolarographic titration. Under an acidic condition, glyphosate can be transferred into a nitrosocompound by reacting with NaNO2. The titration end-point was obtained by the formation of a sharp cut in the oscillopolarographic with infinitesimal NaNO2 on double platinum electrodes. The results showed that glyphosate had an excellent linear relationship over the range of 7.16 x 10(-6) - 4.14 x 10(-3) mol L(-1), the RSD of the proposed method was lower than 3%, and the spiked recoveries of three real water samples were in the range of 97.7 - 100.2%. PMID- 15524194 TI - A hydrogen ion-selective poly(aniline) solid contact electrode based on dibenzylpyrenemethylamine ionophore for highly acidic solutions. AB - Hydrogen-ion selective solid contact electrodes based on tribenzylamine, dibenzylnaphthalenemethylamine, and dibenzylpyrenemethylamine ionophores were prepared. With these electrodes, we showed that the response ranges were influenced by the number of phenyl rings in the ionophores. The lower limits for a linear pH response in acidic solutions were pH 2.50, 0.65, and 0.50, respectively. As the number of phenyl rings in the ionophores increased, the slopes of the EMF responses of these electrodes did not change significantly, but their response extended toward an acidic range (shifted to pH 0.50). Thus, their dynamic response range became wider. A solid contact electrode with dibenzylpyrenemethylamine ionophore, in particular, showed the best selectivity, from the interference of cations and anions, and the best reproducibility of the EMF. This electrode was stored in Tris buffer solutions, artificial serum, and hydrofluoric acid solutions for one month without any loss of performance. Their response time was 8 s. Satisfactory results were obtained when it was tested directly with artificial serum (in pH range 6.0 - 8.5) and hydrofluoric acid. PMID- 15524195 TI - 2,6-bis-methylsulfanyl-[1,3,5]thiadiazine-4-thione as a Ag+-selective ionophore. AB - A silver ion-selective electrode was prepared with a polymeric membrane incorporating 2,6-bis-methylsulfanyl-[1,3,5]thiadiazine-4-thione as an ionophore, tri-n-butylphosphate (TBP) as a plasticizer and sodium tetraphenylborate (NaTPB) as an additive. The electrode exhibited a near-Nernstian response of 52 mV/decade over a wide linear concentration range of 1.0 x 10(-5) - 1.0 x 10(-1) M with a lower detection limit of 9.77 x 10(-6) M. The electrode exhibited excellent selectivity for silver ion over many of the alkali, alkaline-earth and transition metal ions. The electrode worked well over a wide pH range of 1.77 - 7.13. The response time of the electrode was less than 20 s. The sensor can be applied as indicator electrode for the potentiometric titration of Ag+ ions with Cl- ions. PMID- 15524196 TI - An Eu(III) sensor based on N,N-diethyl-N-(4-hydroxy-6-methylpyridin-2 yl)guanidine. AB - A highly selective poly(vinyl chloride)-based membrane sensor produced by using N,N-diethyl-N-(4-hydroxy-6-methylpyridin-2-yl)guanidine (GD) as active material is described. The electrode displays Nernstian behavior over the concentration range 7.0 x 10(-5) - 1.0 x 10(-1) M. The detection limit of the electrode is 5.0 x 10(-5) M. The best performance was obtained with the membrane containing 30% PVC, 55% benzyl acetate, 5% GD and 10% oleic acid. The response of the sensor is pH-independent in the range of 3.0 - 7.0. The sensor possesses satisfactory reproducibility, fast response time (< 20 s), and specially excellent discriminating ability for Eu(III) ion with respect to the alkali, alkaline earth, transition and heavy metal ions. The membrane sensor was used as an indicator electrode in potentiometric titration of Eu(III) ion with EDTA. It was also applied in determination of fluoride ions in mouth wash preparations. PMID- 15524197 TI - Constructing a new optical sensor for monitoring ammonia in water samples using bis(acetylacetoneethylendiamine)-tributylphosphin cobalt(III) tetraphenylborate complex-coated triacetylcellulose. AB - A new ammonia optical sensor was designed using bis(acetylacetoneethylendiamine)tributylphosphin cobalt(III) tetraphenylborate complex, coated on transparent triacetylcellulose film as membrane. The change in the absorbance of the optode at the maximum wavelength of 408 nm was related to ammonia concentration in aqueous samples. A buffer solution with a pH of 9 (sodium borate-HCl) was used. The optode was fully regenerated in pH 2. The linear dynamic range for determination of ammonia was 3.3 x 10(-4) to 6.9 x 10( 3) mol l(-1) with a detection limit of 5.0 x 10(-5) mol l(-1) and a response time range of 4 - 6 min. This membrane was successfully applied for determination of ammonia in drinking water. PMID- 15524198 TI - Flow-through spectrophotometric sensor for the determination of aspartame in low calorie and dietary products. AB - A very simple flow-through sensor is presented for the determination of the intense sweetener aspartame in low-calorie and dietary products. The sensor is implemented in a monochannel flow-injection system with UV spectrophotometric detection using a Sephadex CM-C25 cationic exchanger packed 20 mm high in a flow cell. This method is based on the transient retention of a cationic species of the sweetener on the solid phase when a pH 5.0 acetic acid sodium acetate buffer (0.01 M) is used as a carrier (2.6 mL(-1) min). The carrier itself elutes the analyte from the solid support, regenerating a sensing zone. Aspartame was determined by measuring its intrinsic absorbance at 219 nm at its residence time without any derivatization. Calibration graphs were linear over the range of 5.0 600.0 microg mL(-1) with an RSD of 0.55% (peak height). This sweetener was determined in several samples by measuring the height or peak area, obtaining recoveries ranging between 95 - 101% and 97.5 - 101%, respectively. The procedure was validated for its use in the determination of aspartame in low-calorie and dietary products, giving reproducible and accurate results. PMID- 15524199 TI - Cyanide reaction with ninhydrin: elucidation of reaction and interference mechanisms. AB - A new sensitive spectrophotometric method has recently been developed for the trace determination of cyanide with ninhydrin. Cyanide ion was supposed to act as a specific base catalyst. Nevertheless, this paper demonstrates that the reported assay is based on a novel reaction of cyanide with 2,2-dihydroxy-1,3-indanedione, which affords purple or blue colored salts of 2-cyano-1,2,3-trihydroxy-2H indene. Hydrindantin is merely an intermediary of the reaction. The formation of a stable and isolable ninhydrin-cyanide compound has been confirmed by its preparation in crystalline form. Also, it is thoroughly characterized by elemental as well as MS, IR, UV/VIS and 1H NMR analyses. The Ruhemann's sequence of reactions of cyanide with ninhydrin has been reconsidered and an adequate mechanism of the reaction is proposed. As a consequence, the interference of oxidizers as well as copper, silver and mercury ions with the cyanide determination has been elucidated. PMID- 15524200 TI - Flotation-spectrophotometric determination of mercury in water samples using iodide and ferroin. AB - This paper describes a simple and highly selective method for separation, preconcentration and spectrophotometric determination of trace amounts of mercury. The method is based on the flotation of an ion-associate of HgI4(2-) and ferroin between aqueous and n-heptane interface at pH 5. The ion-associate was then separated and dissolved in acetonitrile to measure its absorbance. Quantitative flotation of the ion-associate was achieved when the volume of the water sample containing Hg(II) was varied over 50 - 800 ml. Beer's law was obeyed over the concentration range of 3.2 x 10(-8) - 9.5 x 10(-7) mol l(-1) with an apparent molar absorptivity of 1 x 10(6) l mol(-1) cm(-1) for a 500 ml aliquot of the water sample. The detection limit (n = 25) was 6.2 x 10(-9) mol l(-1), and the RSD (n = 5) for 3.19 x 10(-7) mol l(-1) of Hg(II) was 1.9%. A notable advantage of the method is that the determination of Hg(II) is free from the interference of the almost all cations and anions found in the environmental and waste water samples. The determination of Hg(II) in tap, synthetic waste, and seawater samples was carried out by the present method and a well-established method of extraction with dithizone. The results were satisfactorily comparable so that the applicability of the proposed method was confirmed in encountering with real samples. PMID- 15524201 TI - Structural features of humic acid of the coastal sediment in Ariake Sea tidelands: use of humic acid as an environmental indicator for river basins and coastal regions. AB - The structural features of humic acid (HA) at the sediment surface of the tideland at the Hayatsuegawa-river mouth at the Ariake Sea were investigated for the utilization of HA toward an environmental indicator of the features of the river basin and coastal region. 1H NMR analysis revealed a high-content hydrocarbon residue with a similar type of terrigenous HA. Direct and methylation pyrolysis-GC analysis suggested the incorporation of long-chain carboxylate in HA in the tidelands. The incorporation of branched-chain carboxylate residues in HA is the result of the microbial decomposition of detritus; these residues could be one of the characteristic structural features of HA in this area, which is rich in biodiversity and microbial activity. Because the structural features of coastal zone HA appear to reveal the characteristics and activities of the biological environment, these findings suggest the possibility of becoming an indicator of the detailed analysis of the structural features of coastal zone HA. PMID- 15524202 TI - Analysis of aliphatic and aromatic carbonyl compounds in ambient air by LC/MS/MS. AB - A sensitive liquid chromatograph/tandem mass spectrometric technique (LC/MS/MS) was applied to determine aliphatic and aromatic carbonyl compounds in ambient air. Traces of the carbonyl compounds were sampled by passing through a Sep-Pak DNPH-silica cartridge. Their derivatives were thus eluted with acetonitrile, separated by reversed-phase liquid chromatography and determined by quadrupole tandem mass spectrometry in an atmospheric pressure chemical ionization (APCI) mode with multiple reaction monitoring (MRM). The detection limits (DL) of the carbonyl compounds were 0.8 - 15 ng/m3. A number of the carbonyl compounds were detected at n.d.- 14 microg/m3 levels. The precursor ion scanning analysis was applied to identify the unknown compounds. PMID- 15524203 TI - Adsorption of divalent transition metal ions with a chelating agent on octadecyl silica gel. AB - This investigation looked at the extraction ability of divalent transition metal ions onto an octadecyl silica gel (C18g) with a 4,4-trifluoro-1-(2-thienyl)-1,3 butadione (TTA) chelating agent. A method of retaining TTA onto C18g (TTA-C18g) was developed in order to adsorb the metal ions. The difference in the half adsorption and half-extraction pH values between transition metals Ni2+-Co2+ was found to be 0.7 in this system. This is better than previously published results of 0.3 for the conventional extraction method using TTA in nitrobenzene. More than 96% of the metal ions in aqueous solution could be adsorbed onto TTA-C18g. Our system, which has no organic phase, can achieve a better removal or separation of transition metal ions than the conventional solvent-extraction methods using TTA in toluene or nitrobenzene. PMID- 15524204 TI - Electrochemical determination of the superoxide ion concentration from KO2 dissolved in dimethyl sulfoxide. AB - An electrochemical determination of the O2- concentration from KO2 in DMSO solution using steady state microelectrode voltammetry shows that the KO2/DMSO method with the combined use of a crown compound or sonication is a reliable and simple technique for introduction of O2- to the biomimitic reaction system. PMID- 15524205 TI - Application of difference NOE-pumping NMR technique and cold-spray ionization mass spectrometry to identify a ligand binding with a protein receptor. AB - A difference diffusion-based NMR technique and cold-spray ionization mass spectrometry were employed as a solution-based approach for identifying a ligand binding with a protein receptor. The difference diffusion-based NMR technique, called difference NOE-pumping, can directly detect the ligand interacting with a protein receptor. This technique uses a simple pulse sequence and the diffusion filter can easily be optimized. The cold-spray ionization mass spectrometry (CSI MS), a variant of electrospray ionization mass spectrometry (ESI-MS) operating at low temperature, has been applied to detect the ligand-receptor complex. The efficiency of these techniques for identifying binding ligands is demonstrated with the human serum albumin (HSA)-drug system. PMID- 15524206 TI - Computational and NMR analyses for the identification of bound water molecules in ribonuclease T1. AB - A structural characterization of bound water molecules in ribonuclease T1 (RNase T1) was carried out by nuclear magnetic resonance spectroscopy and molecular dynamics simulation. Amide protons of residues Trp59, Leu62, Tyr68 and Phe100 were found to cross-relax with protons of bound waters. Molecular dynamics simulations of the 120 water molecules observed in the free form of the crystal structure indicate that these amide protons donate hydrogen bonds to the less mobile water molecules. Hydrogen-bonded chains of the water molecules that are identified in the simulation study are located in the hairpin-like loop of RNase T1, comprising residues 62 to 76. The temperature factors of the observed water molecules in the crystal structure are very low, indicating that these bound waters are intrinsic components of RNase T1. PMID- 15524207 TI - Application of the 19F NMR technique to observe binding of the general anesthetic halothane to human serum albumin. AB - 19F NMR techniques were employed to characterize the binding property of the widely used general anesthetic halothane with human serum albumin (HSA). It was found that 19F(1H) NOE and 2D 1H-19F HOESY experiments detected intermolecular NOEs between halothane 19F and HSA protons. Measurements of the diffusion coefficients for halothane were also carried out by 1H and 19F NMR, indicating the interaction of halothane with HSA. The present results indicate that these techniques are very suitable to identify a fluorine-containing ligand binding with a protein receptor in the drug-discovery process. PMID- 15524208 TI - Oligonucleotides as radiopharmaceuticals. PMID- 15524209 TI - Imaging protein-protein interactions in whole cells and living animals. PMID- 15524210 TI - Radiolabeled peptides in nuclear oncology: influence of peptide structure and labeling strategy on pharmacology. AB - Radiometallo-labeled analogs of SS have shown great benefit in the in vivo localization and targeted radiotherapy of human tumors. The progress and innovation in this clinical application came from the change in strategy, leaving the most widely used radiohalogens for a coordination chemistry approach. The use of chelators appended to the biologically active peptide which convey high thermodynamic and kinetic stability to the radiopeptides did not only improve the pharmacokinetics and pharmacodynamics of the molecules, but surprisingly the biological potency as well. The most urgent problem to be solved in the field is to improve the kidney clearance of the radiopeptides. The kidney turned out to be the dose-limiting organ in this type of targeted radiotherapy. Coordination chemical strategies have already paved the way to a successful clinical application; it is most likely that chelator modification will further help to improve the renal handling of radiometallopeptides. PMID- 15524211 TI - Pretargeted radioimmunotherapy. PMID- 15524212 TI - PET/CT: combining function and morphology. PMID- 15524213 TI - High relaxivity contrast agents for MRI and molecular imaging. PMID- 15524214 TI - Luminescent lanthanide complexes as sensors and imaging probes. PMID- 15524215 TI - Magnetic resonance signal amplification probes. PMID- 15524216 TI - Imaging of proteases for tumor detection and differentiation. PMID- 15524217 TI - Molecular imaging with targeted ultrasound contrast microbubbles. PMID- 15524218 TI - Noninvasive real-time in vivo bioluminescent imaging of gene expression and of tumor progression and metastasis. PMID- 15524219 TI - Targeted optical imaging and photodynamic therapy. PMID- 15524220 TI - Canada or bust? The battle over drug importation. PMID- 15524221 TI - Federal Trade Commission and the Department of Justice come out in favor of competition in health care. PMID- 15524222 TI - Postmenopausal hormone replacement therapy. PMID- 15524223 TI - Childhood tuberculosis in developing countries: prospects for improved diagnosis and control. PMID- 15524224 TI - Significance of recent onset nocturnal enuresis in adult men: a prospective study. AB - INTRODUCTION: Although literature on childhood nocturnal enuresis and its persistence into adulthood is abundant, recent onset nocturnal enuresis in adults is a poorly studied symptom. OBJECTIVE: To determine the significance of recent onset nocturnal enuresis in adult males in relation to lower urinary tract pathology, and its treatment. METHODS: All men with recent onset nocturnal enuresis attending a urology unit over a period of 12 months were evaluated prospectively. Their treatment and outcome were recorded. RESULTS: There were 30 patients (mean age 64 years). Ultrasonography revealed upper urinary tract dilatation in 22 patients. Another six patients had post-void residual urine volume over 500 mL without upper tract dilatation. Only two patients did not have ultrasonographic evidence of bladder outflow obstruction. Elevated blood urea was noted in 14 patients. Twenty patients underwent transurethral resection of the prostate (TURP) and one patient with balanitis xerotica obliterans had circumcision and meatotomy. Three patients were taught clean intermittent self catheterisation, and three patients preferred indwelling urethral catheters. One patient while awaiting TURP died of a myocardial infarction. Eight patients, who were treated with alpha-adrenergic antagonists initially, required further intervention later as the response to medical therapy was poor. CONCLUSIONS: Recent onset nocturnal enuresis in adult males is a symptom closely associated with significant lower urinary tract pathology requiring early urological intervention. Considering its impact on management it is reasonable to classify recent onset nocturnal enuresis as a lower urinary tract symptom in adult men and including it in symptom scores used to assess bladder outflow obstruction. PMID- 15524225 TI - Selection of students for admission to a medical school in Sri Lanka. AB - OBJECTIVES: To assess the extent to which selected entry point factors predicted success in a Sri Lankan medical school. METHODS: The study sample consisted of all students in two consecutive entry cohorts. Marks obtained at the national university entrance examination in physics, chemistry, botany and zoology; the aggregate marks of these four subjects (the only academic criterion used in selection); the district of entry (the other, non-academic criterion); and gender, were identified as entry point variables. Success in a medical school was measured in five ways, including whether a student had passed all examinations in the medical faculty at first attempt or not. Multiple logistic regression was used to assess the extent to which the selected entry point factors could predict variability in outcome measures. RESULTS: Of 331 students, 6.6% were merit quota admissions, and 19.4% were from 'underprivileged' districts; 46.8% were women. Of the entry point factors, being female and obtaining an aggregate of > or =280 (of a possible 400) were the only independent predictors of success in all outcome measures. Obtaining at least grade B in zoology was also an independent predictor of passing all examinations at first attempt. The aggregate score alone accounted for only 2-5% of variance in a medical school performance. There was no association between admission from an underprivileged district and any of the outcome measures. CONCLUSIONS: The one and only measure of academic performance used for selection of students admitted to our medical schools, is a very weak predictor of success in a medical school. PMID- 15524226 TI - Outcome of treatment of multidrug resistant tuberculosis. AB - Multidrug resistant tuberculosis (MDRTB) is a growing problem worldwide. It is an emerging problem in Sri Lanka too although the exact data are not known. This report describes treatment and outcome of 14 MDRTB patients. All have had previous anti-tuberculosis treatment. Out of 32 previous treatment episodes, treatment has been either irregular or defaulted in 26(81%), which has largely contributed to the emergence of drug resistance. Treatment commenced with a combination of second line anti-tuberculosis drugs, namely, amikacin, ciprofloxacin, ethionamide, clofazimine and pyrazinamide. Four patients were considered cured, another four failed treatment, five defaulted treatment and one died. Previous sensitivity to second line drugs and proper isolation facilities were not available. Preparedness to face the threat of MDRTB is essential. Measures should be taken to reduce the rate of defaulters of anti-tuberculosis treatment, and facilities to treat MDRTB should be expanded. PMID- 15524227 TI - Locating an insulinoma by surgical exploration. AB - The case of a 79-year old man who was suspected to have an insulinoma is presented. Although clinically and biochemically an insulinoma was the most probable diagnosis there was no supportive radiological evidence. Open surgery and exploration revealed a lesion which was confirmed to be an insulinoma. The patient's hypoglycaemia improved immediately following surgery. In the Sri Lankan setting where sophisticated imaging procedures are not freely available, open exploration of the pancreas is a good alternative to locate a suspected insulinoma. PMID- 15524228 TI - Pregnancy following treated malignant struma ovarii. PMID- 15524229 TI - A case of pulmonary alveolar microlithiasis. PMID- 15524230 TI - Management of health care waste in Sri Lanka. PMID- 15524231 TI - Medullary carcinoma of the thyroid gland associated with Hashimoto's thyroiditis. PMID- 15524232 TI - Umbilical cord blood lead levels. PMID- 15524233 TI - Delay in investigating haematuria. PMID- 15524234 TI - Losing baby friendliness? PMID- 15524235 TI - Cracked skin of feet: an ignored entity in the tropics. PMID- 15524236 TI - Nutritional status and productivity of Sri Lankan tea pluckers. PMID- 15524238 TI - Turning over a new leaf versus shaking the tree. PMID- 15524242 TI - Aging with multiple sclerosis. AB - Although multiple sclerosis (MS) does not typically reduce life expectancy, there has been relatively little systematic investigation of the experiences and health related concerns of people aging with this disease. A current search of the database CINAHL produced no articles when the search terms "multiple sclerosis," "nursing" and "aging" were used. To initiate more dialogue about the role of nurses in addressing the issues and concerns of people aging with MS, a cross sectional descriptive study was conducted using both qualitative interviews and the administration of standardized instruments to elicit information about the health concerns and service needs of 27 people with MS 55 years of age and older. Qualitatively, participants perceived that they had less freedom and required more assistance than same age peers who do not have MS. Scores from standardized instruments support these perceptions. Participants expressed unmet needs in the areas of housework, physical therapy, MS support groups, religious service attendance, information and referral, check-in services, assistive technology use, social activities, personal care, and care coordination. To address these perceptions and needs, neuroscience nurses need to be aware of and sensitive to the challenges of aging with MS. In addition, nurses must be prepared to discuss and provide information, resources, and referrals on a wide range of health, social, and wellness-related services. PMID- 15524243 TI - Pain in children with cerebral palsy: a review. AB - Children with cerebral palsy (CP) face many challenges including impaired motor control and coordination, functional impairment, sensory disturbances, and, sometimes, communication difficulties and cognitive deficits. Pain also may be a problem for children with CP due in part to the inherent deficits associated with the disease, as well as the invasive medical and surgical procedures and rehabilitative activities children with CP undergo on a regular basis. A review of current literature indicates pain is a common experience for children with CP and has been understudied in this population. Further emphasis and research on appropriate assessment and management strategies sensitive to the unique characteristics and limitations of children with CP are warranted. PMID- 15524244 TI - Neuronal pathway finding: from neurons to initial neural networks. AB - Neuronal pathway finding is crucial for structured cellular organization and development of neural circuits within the nervous system. Neuronal pathway finding within the visual system has been extensively studied and therefore is used as a model to review existing knowledge regarding concepts of this developmental process. General principles of neuron pathway finding throughout the nervous system exist. Comprehension of these concepts guides neuroscience nurses in gaining an understanding of the developmental course of action, the implications of different anomalies, as well as the theoretical basis and nursing implications of some provocative new therapies being proposed to treat neurodegenerative diseases and neurologic injuries. These therapies have limitations in light of current ethical, developmental, and delivery modes and what is known about the development of neuronal pathways. PMID- 15524245 TI - A young onset Parkinson's patient: a case study. AB - Young onset Parkinson's disease (YOPD) is defined as idiopathic Parkinson's disease (IPPD) occurring in people between 21 and 40 years of age; it strikes approximately 5% of Parkinson's patients. YOPD has earlier onset of motor complications than later onset Parkinson's disease. Motor complications and disease progression are responsible for devastating morbidity. Current medical and surgical treatments can dramatically ameliorate motor complications and help maintain function and employment. Patient education, support, and advocacy provided by nursing staff can influence the treatment options for these patients, having a significant effect on the future course of the disease. This case history documents the course of a YOPD patient with unusually severe motor complications. He is the only patient at Puget Sound Neurology ever to develop rhabdomyolysis due to dyskinesias. Following bilateral subthalamic nucleus deep brain stimulation, his Parkinson's symptoms have improved dramatically, and his motor complications are significantly improved. PMID- 15524247 TI - NDT competence of nurses caring for patients with stroke. AB - Neurodevelopmental treatment (NDT) is the most used rehabilitation approach in the treatment of patients with stroke in the Western world today, despite the lack of evidence for its efficacy. The aim of this study was to conduct an intervention check and measure the nurses' competence, in positioning stroke patients according to the NDT approach. The sample consisted of 144 nurses in six neurological wards who were observed while positioning stroke patients according to the NDT approach. The nurses' combined mean competence scores within the wards was 195 (70%) of 280 (100%) possible, and for each ward the mean score varied between 181 (65%) and 206 (74%). This study indicates that nurses working in hospitals where the NDT approach has been implemented have the knowledge and skills to provide NDT nursing. PMID- 15524246 TI - Effect of backrest position on intracranial pressure and cerebral perfusion pressure in individuals with brain injury: a systematic review. AB - Head elevation is a conventional nursing procedure for brain-injured individuals with intracranial hypertension; it is performed with the intent of reducing intracranial pressure (ICP) by means of a noninvasive physical intervention. However, in certain circumstances, head elevation puts the brain-injured individual at risk for secondary cerebral injury because of impaired arterial blood pressure and compromised cerebral perfusion pressure (CPP). A systematic literature search was conducted to evaluate existing evidence regarding the effect of changing the backrest position on ICP and CPP in brain-injured individuals. Eleven articles were retrieved. In nine articles it was concluded that ICP significantly decreased at 30 degrees of head elevation compared with a flat position. Five of the nine articles showed no statistical significance in the magnitude of change in CPP from a flat position to 30 degrees of head elevation. Major limitations in the 11 articles were small sample sizes and unclear study protocols, which may have caused a failure to detect the effect of head elevation. In clinical practice, intensive care unit staff members need to cautiously perform head elevation with a thorough understanding of its physiologic effect and potential hazard. Future research should investigate the effects of therapeutic positions on different neurological and neurosurgical populations and explore the combination of head elevation and lateral side-lying positions. PMID- 15524248 TI - Martin's mantra. Interview by Jennifer Trueland. PMID- 15524249 TI - The evolving nurse. PMID- 15524250 TI - Chilling prospects. PMID- 15524251 TI - A forever kind of caring. PMID- 15524252 TI - Seasons in the sun. PMID- 15524253 TI - Discomfort breaks. PMID- 15524254 TI - Evaluation of a clinical leadership initiative. AB - AIM: To evaluate a clinical leadership initiative in mental health and care of older people settings. METHOD: Clinical leaders (n=15), clinical nurse managers (n=6), and mentors (n=4) involved in the initiative received three questionnaires assessing perceived change, occupational stress and burnout. A nurse, a support worker and a doctor or therapist with whom the clinical leaders worked, each received the perceived change questionnaire (n=45). A comparative group of senior nurses (n=6) who were not part of the initiative received all three questionnaires. A comparative group of clinical nurse managers (n=3) not involved in the initiative received the perceived change questionnaire. RESULTS: Major improvements in direct care management, communication processes, decision-making, clinical nursing care, reporting and evaluation practices, and support strategies and processes at ward level were indicated. The evaluation also identified evidence of stress, potential burnout and job dissatisfaction. CONCLUSION: A customised design was negotiated between the evaluators and the NHS trust, and recommendations were made for wider introduction of the initiative, with enhanced planning, an integrated evaluation process and a systematic approach to job redesign. PMID- 15524256 TI - Evaluating your learning environment. PMID- 15524255 TI - Principles of post-operative patient care. AB - Surgery causes physiological stress on the body and carries inherent risks such as shock and haemorrhage. This article discusses cardiogenic and hypovolaemic shock and outlines the principles of safe and effective post-operative care, including recognising hypovolaemia, maintaining fluid balance and administering pain control. PMID- 15524257 TI - A home of our own. PMID- 15524258 TI - All about job descriptions. PMID- 15524260 TI - "Conscious caring". PMID- 15524259 TI - Are oral weight loss devices within our scope of practice? PMID- 15524261 TI - MDA offers all Blue Cross Blue Shield small group options. PMID- 15524262 TI - Women dentists: the hidden addicts. Health and well-being in the dental profession. PMID- 15524263 TI - Oral lesions of sickle cell anemia: case report and review of the literature. AB - A case report of the oral manifestations of sickle cell anemia is presented, with emphasis on the radiographic features of the disease and the surgical management of periodontal defects. The pathogenesis and clinical course of the disease are discussed, along with a review of clinical characteristics and management protocols. Recommendations for genetic counseling and an update of research in gene therapy of sickle cell anemia are also presented. PMID- 15524264 TI - University of Detroit Mercy: on the move! PMID- 15524265 TI - "The Michigan difference" is under way. PMID- 15524266 TI - [The role of iron in bacterial virulence]. AB - Data on the role of iron in host-bacterium interaction in relation to virulence are summarized. Attention is focused on host iron acquisition pathways in bacteria. Host iron can be acquired by several mechanisms, e.g. from hemoglobin degradation products such as heme and hemin, directly from ferrated transferrin and lactoferrin, indirectly from iron binding proteins by the production of siderophores and from intracellular iron stores (ferritin). Regulation of iron uptake is discussed. PMID- 15524267 TI - [Trends in the incidence of salmonellosis and campylobacteriosis in the Czech Republic]. AB - Salmonellosis and campylobacteriosis are the most frequently reported acute enteric diseases of infectious origin in the Czech Republic. Epidemiological data on salmonellosis and campylobacteriosis have been reportable in this country since 1951 and 1984, respectively. In 2003, 53,486 cases of acute enteric infections were reported: 26,899 (52%) diagnosed as salmonellosis and 20,063 (almost 40%) diagnosed as campylobacteriosis. In 1989, the annual incidence of salmonellosis was three times as high as in the previous year, the upward trend continued until 1995 (528/100,000) and since 1998 the salmonellosis incidence rates have been declining. The incidence of campylobacteriosis showed a progressive increase since 1984 to peak in 2002 with a following slight decline in 2003. Morbidity from salmonellosis and campylobacteriosis is highest in the age group 0-4-year-olds. The most frequent causative agents are Salmonella Enteritidis (96%) and Campylobacter jejuni, respectively. Both infections are foodborne. Ready-to-eat meals, poultry, confectionery and eggs seem to be most frequently implicated in outbreaks of salmonellosis in public catering and families. Sporadic cases of campylobacteriosis are mostly associated with ingestion of poultry and chopped meat. The incidence rates of these two infections are positively correlated with the average daily temperatures. PMID- 15524268 TI - [Surveillance of serious diseases caused by group A streptococci in the Czech Republic in 2003--the Strep-EURO project]. AB - Results obtained in the first year of active surveillance of serious diseases caused by group A streptococci in the Czech Republic carried out within the Strep EURO project are summarized. From January to December 2003, 28 Czech laboratories referred to the National Institute of Public Health 59 group A streptococcal strains isolated from invasive diseases meeting the Strep-EURO definition. At the same time, clinical-epidemiological data and information on the catchment area population were provided. For 2003, the total morbidity and case fatality rates were calculated to be 1.3/100,000 population and 13.5%, respectively. A high proportion (23.7%) of the 59 strains were of type emm 1, initially labeled M1. As many as 24.1%, 17.2% and 20.7% of the strains tested were resistant to erythromycin, clindamycin and tetracycline, respectively. The seemingly lower morbidity rate and higher case fatality rate from serious diseases caused by group A streptococci reported in the Czech Republic for 1994-1996, i.e. 0.4/100,000 and 46.5%, respectively, in comparison with the latest data can be explained by the use of softer international criteria for the Strep-EURO definition of invasive disease. PMID- 15524269 TI - [Antibodies against mosquito-born viruses in human population of an area of Central Bohemia affected by the flood of 2002]. AB - In the Central-Bohemian area affected by the flood of 2002, 497 residents were screened for antibodies against the mosquito-borne viruses Tahyna (TAHV), West Nile (WNV), Sindbis (SINV) and Batai (BATV; syn. Calovo) using the haemagglutination-inhibition (HIT) and plaque-reduction neutralization (PRNT) tests. Blood samples were collected in September 2002 when the mosquito populations showed the maximum density following the flood. Antibodies against TAHV (16.5% persons in PRNT, 14.9% in HIT), SINV (1.4% in HIT) and BATV (1.4% in HIT, 0.2% in PRNT) were detected. Although 6.8% and 1.2% of the subjects tested reactive with WNV in HIT and PRNT, respectively, the results were interpreted as cross-reactivity with tick-borne encephalitis virus. The seroprevalence of TAHV (both in PRNT and HIT) showed no association with gender (15.8% of males, 16.9% of females), increased with age (1.4% of persons younger than 20 years, 11.2% of persons aged between 20 and 50 years, and 26.2% of persons older than 50 years were positive), and correlated with the mosquito peri-residential challenge (5.0% residents seropositive in a mosquito-free control zone D--mostly Prague, 14.7% in a mild-risk zone C, 20.5% in a moderate-risk zone B, and 28.0% in the most heavily mosquito-infested risk zone A). The highest TAHV seropositivity rate (> 25%) was found amongst the inhabitants of the villages Obristvi, Kozly, Tuhan, Chrast, Chlumin and Hostin. Paired blood samples were obtained from 150 of the persons at a 6-month interval: an infection episode with TAHV during or after the flood was clearly evidenced in one person living in Obristvi, and less convincing findings of recent TAHV infections were found in other three residents of Chlumin and Obristvi (seroconversion and/or significant antibody titres increase detected in HIT only). This serosurvey indicated the existence of an active natural focus of Valtice fever (TAHV infection) stretched along the river Labe nearby Neratovice (Obristvi, Chlumin, Tuhan; Kozly, Tisice, Chrast), and a low TAHV activity area along the lower reaches of the river Vltava between Zloncice and Bukol/Zalezlice. An increased population density of mosquitoes after the flood may have boosted the incidence of mosquito-borne virus diseases, particularly Valtice fever, in Central Bohemia. An optimum prophylactic strategy to control these diseases would be epidemiological surveillance (including monitoring of both the density of mosquitoes and their rate of infection with viruses in natural foci) on the basis of which antiepidemic measures such as integrated mosquito control can be taken. PMID- 15524270 TI - [Direct detection of Treponema pallidum in diagnosis of syphilis]. AB - Available methods for direct diagnosis of syphilis are summarized with emphasis being on those promising for routine use. Direct detection of the causative agent T. pallidum is limited since the agent is not able to synthesize enzyme cofactors, fatty acids and nucleotides de novo, is completely dependent on its host and thus culture on synthetic media is not feasible. Direct diagnosis of syphilis is based on rabbit infectivity testing (RIT), dark field or fluorescent microscopy and recently also on molecular biological methods used with increasing frequency in routine practice. Suitability and usability of different methods for direct detection of T. pallidum at different stages of syphilis are explained. Except for molecular biological methods, most of detection techniques can only be used at the primary and secondary stages or in early congenital syphilis. Major PCR methods for diagnosis of syphilis are presented. Not all of them are suitable for use in routine practice owing to differences in their sensitivity and design. The polA PCR method appears to be the most promising in this regard. PMID- 15524271 TI - [Problems related to control of epidemic and persistent outbreaks of scabies]. AB - Experience with control of epidemic and persistent outbreaks of scabies in collectives is presented. Both diagnosis and patient's compliance may pose problems. When diagnosing scabies, not only clinical symptoms but also the dynamics of the spread among persons in collective facilities such as social care and psychiatric in-patient centres should be taken into account and efficacy of the antiscabies medication should be followed up. If diagnosis of scabies is confirmed, it is crucial to prescribe an effective antiscabies medication and to ensure the patient's compliance with treatment, including thorough washing of the body each time before application of the medication, throughout the whole treatment period instead of treating only the symptoms of secondary eczema. PMID- 15524272 TI - [Antibodies against low density oxidized lipoproteins in type 2 diabetics]. AB - Oxidative modification of low density lipoproteins (LDL) is an important factor in the development of macrovascular atherosclerotic complications in patiens with type 2 diabetes mellitus. Recently autoantibodies against oxidized LDL (anti oxLDL) have been suggested as a potential marker of LDL oxidation in vivo. The purpose of this study was to investigate the presence and levels of anti-oxLDL in patients with type 2 diabetes compared to healthy persons. We determined the serum concentrations of anti-oxLDL in 20 type 2 diabetic patiens with different degree and type of atherosclerotic vascular damage. Two healthy population groups: 20 young blood donors and 20 age and gender matched persons were used as controls. Anti-oxLDL positivity rates were distinctively higher in both control groups. Concentrations of anti-oxLDL were significantly lower in diabetic patients compared to both control groups. The incidence rates and levels of anti oxLDL in both control groups were similar. Anti-oxLDL levels in the diabetes group did not correlate with the degree of macrovascular damage, serum total cholesterol, LDL cholesterol and triglyceride concentrations. We did not find any significant relationship between anti-oxLDL and other oxidative stress factors (superoxide dismutase, malondialdehyde, C and E vitamins). We suppose that anti oxLDL may have an antiatherogenic protective role in healthy people but are not applicable to be an in vivo marker of LDL oxidation and macrovascular atherosclerotic vascular damage. PMID- 15524273 TI - [Host immune response in mammals to Encephalitozoon cuniculi infection]. AB - The immunity system status of an individual plays the key role in regulation of opportune infection. In the fight against the intracellular parasites several non specific as well as specific immunity mechanisms are applied. The dominant role in response to infection caused by the representatives of genus Encephalitozoon plays the cell-mediated immune response. Encephalitozoon cuniculi, as the most explored representative of this genus is able to survive in the host organism despite his active immunity response. Latent asymptomatic infection goes on only as long as the parasite multiplication and immune response are balanced. PMID- 15524275 TI - An expression profile of human alpha-lactalbumin in the milk of transgenic mouse. AB - Five female transgenic mice were produced by microinjection using a construct made up of a 7.3-kb-5' flanking region and a 2.0-kb coding region of human alpha lactalbumin, as well as a 227-bp 3'-flanking region from bovine growth hormone gene. A founder female expressed human alpha-lactalbumin as much as 0.3 g per liter of its milk, approximately a 3-fold increase in the total alpha-lactalbumin concentration of the transgenic mouse milk. Compared with the normal mice, the expression profile of the halpha-Lac transgene in the transgenics is different during the lactation, showing low level in the first 3 days and becoming increased from day 4, then gradually reaching and stabilizing at the highest level from day 13. In addition, the milk yielding volume in the transgenics tended to be higher than in normal mice, suggesting higher concentrations of alpha-lactalbumin might boost more milk output. PMID- 15524276 TI - Effects of clinorotation on COL1A1- EGFP gene expression. AB - Bone-formation related gene plays a critical role in bone loss induced by space microgravity, however the exact mechanism is unclear. In this study, we aim to investigate the effect of microgravity on the activity of alpha 1(I) collagen (COL1A1) gene promoter and the expression of osteoblast-related genes. COL1A1 promoter was digested by restriction enzymes resulting in three DNA fragments. The fragments were ligated with the enhanced green fluorescent protein report gene, and subcloned into expression vectors. ROS17/2.8 cells transfected by these vectors were screened by G418, and enhanced green fluorescent protein (EGFP) positive colonies were isolated and cultured under clinostat condition. EGFP and Collagen type I expression level were detected by fluorescence intensity analysis and immunocytochemistry methods respectively. The results showed that the expression of EGFP and collagen type I was increased 24 h, 48 h after the cells were cultured under stimulated microgravity, illustrating that the activity of COL1A1 promoter might be increased. In conclusion, osteoblasts can compensatively increase the expression of type I collagen by enhancing the activity of COL1A1 promoter under short-term simulated microgravity conditions. PMID- 15524277 TI - Establishment and application of minigene models for studying pre-mRNA alternative splicing. AB - The objective of the present study is to establish a minigene model for studying pre-mRNA alternative splicing. To prepare the minigene DNA constructs, with human or mouse genomic DNA as templates, GluR-B, FGF-2R and Zis "minigene" fragments were amplified using PCR and cloned to the eukaryotic expression vectors. The three constructed minigenes and the expression vectors of Tra2beta1 and Zis2 were co-transfected in Hela cells. RT-PCR analysis was performed to semi quantitatively determine the spliced products from the minigenes. The results demonstrated that the constructed minigenes are useful in studying the pre-mRNA alternative splicing in cultured cells. With the established Zis minigene, we for the first time found that Zis2 isoform regulates the alternative splicing of Zis minigene. PMID- 15524278 TI - Discovery of laryngeal carcinoma by serum proteomic pattern analysis. AB - Laryngeal carcinoma is the most common malignancy among head and neck tumors. The purpose of this study is to find biomarkers for laryngeal carcinoma in patient blood serum using the Surface Enhanced Laser Desorption/Ionization (SELDI) technique. Serum samples from 33 laryngeal carcinoma (12 cases of glottis, 18 of supraglottis and 3 of subglottis) patients and 31 age- and sex-matched healthy people were analyzed by SELDI-TOF on a ProteinChip reader, PBSII-C. Protein profiles were generated using WCX2 protein chips. Protein peak clustering and classification analyses were performed utilizing the Biomarker Wizard and Biomarker Pattern software packages, respectively. The results showed that sixteen peaks had significant difference between laryngeal cancer patients and healthy group, eight of which were up-regulated in the patient samples, and the others were down-regulated. Two protein peaks 8153 Da and 2035 Da were automatically chosen for the system training and development of a classification tree. The analysis yielded a correct percentage of 96.9% for patients and 96.7% for control. The results suggest that serum is a useful resource for the detection of specific biomarkers for laryngeal carcinoma. Proteinchip Array System was a useful tool for a high throughput screening of large-sized serum samples to discover potential biomarkers for carcinoma. PMID- 15524279 TI - Three-dimensional structure determination of capsid of Aedes albopictus C6/36 cell densovirus. AB - The three-dimensional structure of capsid of Aedes albopictus C6/36 densovirus was determined to 14-A resolution by electron cryomicroscopy and computer reconstruction. The triangulation number of the capsid is 1. There are 12 holes in each triangular face and a spike on each 5-fold vertex. The validity of the capsid and nucleic acid densities in the reconstructions was discussed. PMID- 15524280 TI - Increasing terrestrial vegetation activity in China, 1982-1999. AB - Variations in vegetation activity during the past 18 years in China were investigated using the normalized difference vegetation index (NDVI) derived from the 3rd generation time series dataset of NOAA-AVHRR from 1982 to 1999. In order to eliminate the effects of non-vegetation factors, we characterized areas with NDVI < 0.1 as "sparsely vegetated areas" and areas with NDVI > or = 0.1 as "vegetated areas". The results showed that increasing NDVI trends were evident, to varying extents, in almost all regions in China in the 18 years, indicating that vegetation activity has been rising in recent years in these regions. Compared to the early 1980s, the vegetated area increased by 3.5% by the late 1990s, while the sparsely vegetated area declined by 18.1% in the same period. The national total mean annual NDVI increased by 7.4% during the study period. Extended growing seasons and increased plant growth rates accounted for the bulk of these increases, while increases in temperature and summer rainfall, and strengthening agricultural activity were also likely important factors. NDVI changes in China exhibited relatively large spatial heterogeneity; the eastern coastal regions experienced declining or indiscernibly rising trends, while agricultural regions and western China experienced marked increases. Such a pattern was due primarily to urbanization, agricultural activity, regional climate characteristics, and different vegetation responses to regional climate changes. PMID- 15524281 TI - Differentiation potential of bone marrow mesenchymal stem cells into retina in normal and laser-injured rat eye. AB - Bone marrow mesenchymal stem cells (MSCs) can develop into hematopoietic and mesenchymal lineages but have not been known to participate in the production of retina. Here we report that bone marrow mesenchymal stem cells, after being subretinally transplanted into normal or Nd: YAG laser-injured rat eye, can integrate into RPE layer, photoreceptor layer, bipolar cell layer and ganglion layer. DAPI-labeling detection was used to trace the origin of the repopulating cells. DAPI fluorescence was used to identify retina cells of bone marrow origin 10, 20, 35 and 50 days after transplantation. No formation of rosettes was found but some random cells were found at the end of the observation. MSCs-originated cells spread more widely in the injured retinas than in the normal ones. Immunohistochemical detection showed that though the cells could express neuronal nuclei (NeuN), neuron specific enolase (NSE), glial fibrillary acidic protein (GFAP) and cytokeratin (CK), the proteins expression in the injured transplantation group was abnormal in some region compared with that in the normal transplantation group. Electroretinogram (ERG) showed that ERG-b wave of the injured transplantation group is significantly higher than that of the two laser-injured control groups. These results suggest that a proportion of MSCs can differentiate into retina-like structure in vivo and the differentiation differs in normal and laser-injured retinas. PMID- 15524282 TI - The effect of cdk-5 overexpression on tau phosphorylation and spatial memory of rat. AB - In Alzheimer's disease (AD), hyperphosphorylation of tau may be the underlying mechanism for the cytoskeletal abnormalities and neuronal death. It was reported that cyclin-dependent kinase5 (cdk-5) could phosphorylate tau at most AD-related epitopes in vitro. In this study, we investigated the effect of cdk-5 overexpression on tau phosphorylation and spatial memory in rat. We demonstrated that 24 h after transfection into rat hippocampus, cdk-5 was overexpressed and induced a reduced staining with antibody tau-1 and an enhanced staining with antibodies 12e8 and PHF-1, suggesting hyperphosphorylation of tau at Ser199/202, Ser262/356 and Ser396/404 sites. Additionally, the cdk-5 transfected rats showed long latency to find the hidden platform in Morris water maze compared to the control rat. 48 h after transfection, the level of cdk-5 was decreased significantly, and the latency of rats to find the hidden platform was prolonged. It implies that in vivo overexpression of cdk-5 leads to impairment of spatial memory in rat and tau hyperphosphorylation may be the underlying mechanism. PMID- 15524283 TI - Nuclear translocation of EGF receptor regulated by Epstein-Barr virus encoded latent membrane protein 1. AB - Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP1) is considered to be the major oncogenic protein of EBV encoded proteins, and also it has always been the core of the oncogenic mechanism of EBV. Traditional receptor theory demonstrates that cell surface receptors exert biological functions on the membrane, which neither enter into the nucleus nor directly affect the transcription of the target genes. But, advanced studies on nuclear translocation of the epidermal growth factor receptor (EGFR) family have greatly developed our knowledge of the biological function of cell surface receptors. In this study, we used Tet-on LMP1 HNE2 cell line as a cell model, which is a dual-stable LMP1 integrated NPC cell line and the expression of LMP1 in which could be regulated by Tet system. We found that LMP1 could regulate the nuclear translocation of EGFR in a dose-dependent manner from both quantitative and qualitative levels through the Western blot analysis and the immunofluorescent analysis with a laser scanning confocal microscope. We further demonstrated that the nuclear localization sequence of EGFR played some roles in the location of the protein within the nucleus under LMP1 regulation, and the nuclear accumulation of EGFR regulated by LMP1 was in a ligand-independent manner. These findings provide a novel view that the regulation of LMP1 on the nuclear translocation of EGFR is critical for the process of nasopharyngeal carcinoma. PMID- 15524284 TI - Phylogenetic clusters of rhizobia revealed by genome structures. AB - Rhizobia, bacteria that fix atmospheric nitrogen, are important agricultural resources. In order to establish the evolutionary relationships among rhizobia isolated from different geographic regions and different plant hosts for systematic studies, we evaluated the use of physical structure of the rhizobial genomes as a phylogenetic marker to categorize these bacteria. In this work, we analyzed the features of genome structures of 64 rhizobial strains. These rhizobial strains were divided into 21 phylogenetic clusters according to the features of genome structures evaluated by the endonuclease I-CeuI. These clusters were supported by 16S rRNA comparisons and genomic sequences of four rhizobial strains, but they are largely different from those based on the current taxonomic scheme (except 16S rRNA). PMID- 15524285 TI - Identification of binding epitope of a monoclonal antibody (Z12) against human TNF-alpha using computer modeling and deletion mutant technique. AB - The genes of the heavy and light chain variable region (VH, VL) of Z12 antibody against hTNF-alpha were cloned, and according to the translated sequence of amino acids, the spatial structures of VH and VL domains were modeled by using homology based modeling method, followed by constructing the whole three-dimensional structure of Fv fragment. The complex model of Fv interacting with hTNF-alpha was gained with computer-guided molecular docking method, based on which, it was predicted that the epitope recognized by Z12 was from 141 to 146 of hTNF-alpha. hTNF-alpha molecule was divided into two fragments of N-terminal region from 1 to 91 and C-terminal region from 92 to 157 with prokaryotic expression. The measured results suggested that the antigenic epitope recognized by Z12 antibody was located in the C-terminal region 92-157 of hTNF-alpha, proving the predicted result reliable preliminarily. Further experimental results showed that after hTNF-a 141-146 residues were deleted, Z12 antibody almost lost the ability to recognize the mutant, suggesting that the amino acid residues from 141 to 146 of hTNF-alpha were specially recognized by Z12 antibody. PMID- 15524286 TI - Single nucleotide polymorphism analysis on melanocortin receptor 1 (MC1R) of Chinese native pig. AB - Melanocortin receptor 1 (MC1R) gene, one of the important candidate genes for coat color trait, was used to analyze the single nucleotide polymorphism (SNP) in Chinese native pig breeds by PCR-single strand conformation polymorphism (PCR SSCP). The study had also taken 3 imported pig breeds as control. The results showed that the three mutations G284A, T309C and T364C found in Chinese native pigs were consistent to the mutation found in the European Large Black individuals. However, 68CC or C492T and G728A were only found in the imported individuals, which were obviously different from the Chinese native pigs. Accordingly, we presumed that the coat colors of Chinese native pigs belonged to dominant black color system, which was completely distinct to that of imported pig breeds. Thus it was implied that MC1R gene was not the principal factor affecting the coat color differences of Chinese native pig breeds, but could be used to trace the molecular evolution of pig breeds. PMID- 15524287 TI - [Research of GAP implement supporting system of Chinese medicine plant]. AB - The paper presented a total system frame of supporting GAP implement of Chinese medicine plant in which information technology played an important role. A supporting system of GAP implement have been demonstrated on the GAP base in Zhangshu, Jiangxi. PMID- 15524288 TI - [Determination of hyperin in genus Hypericum medicinal plants]. AB - OBJECTIVE: To compare the content of hyperin in Hypericum medicinal plants. METHODS: HPLC method was established. Alltima C18 radial compression column was used with a mobile phase of methanol-0.5% phosphoric acid (45:55, adjusted to pH = 3.0 with triethylamine). The detecting wavelength was 360 nm. Flow rate was 0.8 ml/min, external standard method was quantitative analysis method. RESULTS: The linear range was 0.101-2.023 microg. The average recovery was 96.527%. The result showed that the content of hyperin in H. perforatum, H. forrestii was higher than that of others (1.257% and 0.461% respectively). CONCLUSION: This method provides an accurate and sensitive way in detecting hyperin. PMID- 15524289 TI - [Microscopic identification of commercial Herba Cistanches by digital imaging technique]. AB - OBJECTIVE: To identify the commercial drugs of Herba Cistanches collected from 10 different areas. METHOD: Descriptions identification and digital imaging technique. RESULT: The original plants of the commercial drugs were three species, Cistanche deserticola Y. C. Ma, C. salsa (C. A. Mey) Benth et Hook. f. and C. tubelosa (Schrenk.) R. Wight. CONCLUSION: The confusion of Herba Cistanches in markets is exist. The main current species was C. deserticola, C. salsa (each 2/5) and C. tubelosa (1/5). The descriptions of C. deserticola and C. salsa are similar and not easy to distinguish. But the digital photographs offered by the paper visually reflected the microscopic differences of two species. PMID- 15524290 TI - [Identification of Fructus Schisandrae chinensis and Fructus Schisandrae sphenantherae in wu zi yan zong pill by RP-HPLC analysis]. AB - OBJECTIVE: To identify Fructus schisandrae chinensis and Fructus schisandrae sphenantherae in wu zi yan zong pill (WZYZP). METHODS: Columm SUPELCOSIL C18; Mobile phase: methanol-water (65:35); Detection wavelength: 254 nm. RESULTS: Using above optimum chromatography conditions, the HPLC chromatography of relative medicinal materials and WZYZP was established. CONCLUSION: HPLC chromatography can be used for identification of Fructus schisandrae chinensis and Fructus schisandrae sphenantherae in WZYZP. PMID- 15524291 TI - [Separation and identification of two benzotropolones from teapigment]. AB - OBJECTIVE: To study the chemical constituents of teapigment. METHOD: Some constituents were isolated by chromatographic methods and identified by chemical constituents and their structures were elucidated by spectral data. RESULT: From teapigment, two compounds have been isolated and identified as purpurogallin (I) and purpurogallin carboxylic acid (II). CONCLUSION: These compounds were main pigments obtained from teapigment for the first time, and likely to be active compounds of teapigment. PMID- 15524292 TI - [Studies on chemical constituents from Polygonum macrophyllum]. AB - OBJECTIVE: To study the chemical structures from Polygonum macrophyllum D. Don. METHODS: To isolate the constituents by reverse phase chromatography and characterize their structures by the analysis of chemical property and spectral data. RESULT: Six compounds were isolated from the 70% acetone extract from the complete herb of P. macrophyllum. Their structures were elucidated as (-) epicatechin-5-O-beta-D-glucopyranoside (I), (+)-catechin-7-O-beta-D glucopyranoside (II), 1-(3-O-beta-D-glucopyranosyl 4,5-dihydroxy-phenyl)-ethanone (III) (-)-epicatechin (IV), chlorogenic acid (V) and gallic acid (VI). CONCLUSION: Compounds I-VI were isolated from the plant for the first time. PMID- 15524293 TI - [Isolation of plant insulin from Momordica charantia seeds by gel filtration and RP-HPLC]. AB - Hypoglycemic polypeptide (PA) was extracted from Momordica charantia seeds with organic acid and ethanol and purified with Sephadex G-50 gel filtration and RP HPLC. PA was judged as plant insulin on the base of the analysis of its SDS-PAGE electrophoresis and amino acid composition. PMID- 15524294 TI - [Inhibition of proliferation and influence of proto-oncogenes expression by matrine in C6 cell]. AB - OBJECTIVE: This study were designed to investigate the inhibition of matrine on C6 glioma cell line and its mechanism. METHODS: MTT was used to measure the levels of the proliferation of C6 cultured with matrine in different concentrations. The effects of matrine on cell cycle of C6 were observed by FCM. The expression of proto-oncogenes C-myc was measured by RT-PCR. RESULT: The proliferation of C6 was obviously inhibited by matrine in a dose dependent manner. The inhibitory rate came to 53.8% +/- 5.6%, when cultured with matrine at 0.20 g/L. The outcome of FCM showed that the proportion of G0/G1 phase cells was decreased obviously, the proportion of S phase cells was reduced when cultured with matrine at 0.20 g/L in 3 days. The outcome of RT-PCR showed that the expression of proto-oncogene C-myc was notably decreased when the dose of matrine was increased. CONCLUSION: Matrine can inhibit the proliferation of C6 and inhibit the expression of proto-oncogenc C-myc. PMID- 15524295 TI - [The inhibitive effects of the ethanol extract from Radix et Rhizoma Rhei on the secretion of HBsAg and HBeAg]. AB - OBJECTIVE: To study the anti-HBV effects of the ethanol extract from Radix et Rhizoma Rhei. METHODS: The influence of the ethanol extract from Radix et Rhizoma Rhei on the secretion of HBeAg and HBsAg was observed through the culture of the 2.2.15 cell with the ethanol extract. RESULTS: 11 days after the ethanol extract's action on the 2.2.15 cell, its 50% concentration dose (CD50) is 39.69 g/L; inhibiting dose (ID50) to HBsAg and HBeAg are 3.29 g/L and 2.34 g/L respectively, and TI 12.06 and 16.96 respectively. CONCLUSION: The ethanol extract from Radix et Rhizoma Rhei can markedly inhibit the secretion of HBsAg and HBeAg in 2.2.25 cell lines. PMID- 15524296 TI - [Therapeutic effects of a low molecular weight peptide from cattle spleen on masugi nephritis in rats]. AB - OBJECTIVE: To investigate the therapeutic effects of a low molecular weight peptide KZ-36 from cattle spleen on masugi nephritis in rats. METHODS: The rabbit serum of anti-rat kidney was initially prepared, and then injected into normal rats to induce the formation of masugi nephritis. In this model, KZ-36 was administrated (i.p.) at the dosage of 50 microg/kg/d for 2 weeks, while saline and hydrocortisone were respectively used as negative and positive controls. The pharmacodynamic parameters were collected. RESULTS: After KZ-36 administration for one week, the protein output in 24 h urine remarkably decreased, and was 5.0 +/- 4.1 mg for two weeks, which was much less than 17.2 +/- 14.5 mg, a counterpart in saline control (P<0.01). KZ-36 could also reduce the serum level of creatinine and improve the renal pathological injury. CONCLUSION: KZ-36 has obvious therapeutic effects on masugi nephritis in rats. PMID- 15524297 TI - [The anti-respiratory syncytial virus (RSV) effect of Radix Glycyrrhizae in vitro]. AB - OBJECTIVE: To develop safe and effective anti-RSV new medicine from Radix Glycyrrhizae. METHOD: The anti-RSV effect of Radix Glycyrrhizae in Hela cell culture was observed by means of the inhibition of cytopathic effect. RESULT: In Hela cell culture, Radix Glycyrrhizae was found to be a inhibitor of RSV in a concentration-dependent manner. The median toxic concentration (TC50) of Radix Glycyrrhizae was 3.43 g/L, the median effective concentration (EC50) of Radix glycyrrhizae against replication of the Long strain of RSV in Hela cells were 0.2535 g/L, the selectivity index (TI = TC50/EC50) is 13.53. In time of addition experiment, Radix Glycyrrhizae inhibited the effect of RSV in Hela cells when it was added at 0 h, 2 h, 4 h, 6 h, 8 h after virus infection. CONCLUSION: In Hela cell culture, Radix Glycyrrhizae was found to be a inhibitor of RSV, there are many ways in the mechanisms. PMID- 15524298 TI - [Protective effect of ganfukang capsule on hepatocytes in rats with experimental hepatic fibrosis]. AB - OBJECTIVE: To observe the protective effect of ganfukang capsule on hepatocytes of rats with experimental hepatic fibrosis rat. METHOD: SD rat model of liver fibrosis was induced by CCl4. The effect of ganfukang capsule on liver function, histological change and ultrastructural were examined. RESULTS: Liver function, histological change and ultrastructural were significantly improved. The degree of steatosis was significantly decreased. CONCLUSION: Ganfukang capsule might protect hepatocytes, mitochondria from the injury induced by CCl4 and improve liver function. PMID- 15524299 TI - [Experimental study on treatment of minimal change nephropathy with niaozhuoqing]. AB - OBJECTIVE: To study the therapeutic effect of niaozhuoqing on minimal change nephropathy (MCN) in rats. METHODS: The MCN rat model was induced by intravenous injection of adriamycin. The effect of treatment on 24h urine protein, serum protein, lipid metabolism, malondialdehyde (MDA), superoxide dismutase (SOD) in serum, and renal histopathological changes were observed. RESULTS: Niaozhuoqing could decrease the contents of proteinuria, blood lipid and MDA in serum, increase serum protein, SOD level and pathologically restore the glomeruli. CONCLUSION: Niaozhuoqing has good effects on MCN, which probably related to decrease serum lipids level and reduce oxygen free radicals. PMID- 15524300 TI - [Pharmacological mechanism of Semen Litchi on antagonizing insulin resistance in rats with type 2 diabetes]. AB - OBJECTIVE: To investigate the pharmacological mechanism of Semen Litchi water extract (SL) on enhancing insulin sensitivity in type 2 diabetic rats (T2DR) with insulin resistance (IR). METHODS: The effects of SL were observed on serum contents of fasting glucose (FSG), blood urea nitrogen (BUN), creatine (Cr), total proteins (TP), albumin (A), malondialdehyde (MDA), total cholesterols (TC), triacylglycerol (TG), free fatty acid (FFA), leptin, tumor necrosis factor-alpha (TNF-alpha), fasting insulin (Fins) and index of insulin sensitivity (ISI), and activities of superoxide dismutase (SOD), alanine aminotransferase (ALT), aspartate aminotransferase (AST) in T2DR-IR. RESULTS: In T2DR-IR, SL could lower concentrations of FSG, TC, TG, FFA, leptin, TNF-alpha and Fins (P < 0.05-0.01), increase ISI (P < 0.01), reform the hyperinsulinemia and insulin sensitivity, decrease the levels of BUN, Cr and the activities of ALT, AST, increase concentration of TP, A (P < 0.05-0.01), recover the hepatic and nephric functions, increase the activity of SOD and decrease content of MDA (P < 0.01). CONCLUSION: SL could reduce the levels of TNF-alpha, hyper-leptinemia and hyperinsulinemia, antagonize insulin resistance, fortify insulin sensitivity, readjust lipodystrophy and maladjustment of glycometabolism, enhance antioxidation,and improve functions of liver and kidney in T2DR-IR. PMID- 15524301 TI - [Studies on the analgesic and anti-inflammatory effect of bornyl acetate in volatile oil from Amomum villosum]. AB - OBJECTIVE: To study analgesic and anti-inflammatory effects of bornyl acetate, the main ingredient of Amomum villosum volatile oil. METHODS: The analgesic effects were tested by hot-plate and writhing reaction method, the ear swelling caused by dimethylbenzene in mice. RESULTS: Bornyl acetate could restrain writhing reaction caused by acetic acid glacial, lighten the pain caused by hot plate. It could also suppress ear swelling caused by dimethylbenzene in mice. CONCLUSION: Bornyl acetate shows analgesic and anti-inflammatory effects. PMID- 15524302 TI - [Experimental study on inclusion compound of beta-cyclodextrin with limonene]. AB - OBJECTIVE: To study the inclusion action of beta-cyclodextrin (beta-CD) with limonene. METHOD: Orthogonal design. RESULT: The optimum preparing condition was a proportion of 1:9 (ml:g) for oil to beta-CD inclusion, temperature at 40 degrees C and stirred for 2 h. CONCLUSION: The condition was optimum for preparation of the inclusion compound of beta-CD with the volatile oil of limonene. PMID- 15524303 TI - Restoring the relationship: the key to nurse and patient satisfaction. AB - Want to improve staff and patient satisfaction scores? Focus on the hospital's most important connection--the bond between nurses and patients. PMID- 15524304 TI - Exercising ethical leadership: conflicts of interest. AB - In this excerpt from a recently published AHA book, the authors provide direction on how to deal with conflicts of interest within a framework you can use for other ethical problems. PMID- 15524305 TI - Trustee certification: around the corner or miles away? AB - How do lay boards make tough decisions on complicated hospital issues? The verdict is still out on whether mandatory trustee education is the best solution, but you need to confront the pros and cons before the choice is made for you. PMID- 15524306 TI - The challenge of emergency departments: what boards can do. PMID- 15524307 TI - Strategic planning: what kind of CEO will your hospital need next? A model for succession planning. PMID- 15524308 TI - Phylogeographic analysis of the firefly, Luciola lateralis, in Japan and Korea based on mitochondrial cytochrome oxidase II gene sequences (Coleoptera: Lampyridae). AB - Luciola lateralis is widely distributed throughout the Korean Peninsula, northeast China, Sakhalin, and Japan. Two ecological types are recognized in Japan based on flash and hatching time characteristics. The mitochondrial cytochrome oxidase II gene was surveyed by restriction fragment length polymorphism analysis for Japan (46 populations) and Korea (two populations). Eleven haplotypes were detected. Gene trees revealed that haplotypes between Japan and Korea are much more differentiated in nucleotide sequences (8.1%) than those within Japan (0.3-1.4%) and Korea (0.7%). Haplotypes between Honshu and Hokkaido are not separated as clades, and the two ecological types cannot be segregated from each other phylogenetically. We suggest that the Japanese populations of this species may have dispersed within one million years ago and that ecological differences may be the result of physiological adaptation to cold climates. PMID- 15524309 TI - Phylogenetic analysis of Mediterranean Mugilids by allozymes and 16S mt-rRNA genes investigation: are the Mediterranean species of Liza monophyletic? AB - The family Mugilidae (Pisces, Mugiliformes) includes species which are present in all tropical and temperate regions. Six species, Chelon labrosus, Mugil cephalus, Liza aurata, L. ramada, L. saliens, Oedalechilus labeo, are commonly found in the Mediterranean. These species have been widely studied through morphological, biochemical, and molecular markers. However, their phylogenetic relationships, and therefore the assumed monophyly of Liza species, still remain unclear: To further investigate this topic, gene-enzyme systems and sequences of the partial 16S rRNA mitochondrial gene were analyzed in Italian samples of all six Mediterranean species. The phylogenetic reconstructions indicated M. cephalus as being the most divergent species and the existence of a main cluster including all the Mediterranean species of Liza and C. labrosus. The parametric bootstrap approach adopted to test alternative phylogenetic hypotheses indicated that the Mediterranean species of Liza do not form a monophyletic group exclusive of Chelon. PMID- 15524310 TI - Biochemical polymorphisms and genetic relationships in rodents of the genera Oryzomys and Oligoryzomys (Sigmodontinae) from Brazil. AB - An electrophoretic analysis of 12 allozyme systems (14 loci, 40 alleles) was performed in the rodent genera Oryzomys and Oligoryzomys, in order to determine the levels of genetic variability within and among populations and species. One hundred and fifty-five individuals from 16 populations of Oryzomys russatus, Or. angouya, Oligoryzomys flavescens, and O1. nigripes species were trapped in nine Brazilian localities. Genetic divergence among populations, as well as the interpopulational gene diversity, was higher in Oryzomys than in Oligoryzomys. The dendrogram of the phenetic relationships among the 16 populations of these four species displays three clusters: the first one joins the O1. flavescens populations with those of O1. nigripes; the second groups the populations of Or. russatus, and the last assembles the populations of Or. angouya. The genetic parameters analyzed reveal that the species belonging to the genus Oryzomys are genetically more structured than those of Oligoryzomys, the latter presenting lower levels of intrademe genetic differentiation (G(ST)') PMID- 15524311 TI - Phylogenetic relationships of 12 penaeoidea shrimp species deduced from mitochondrial DNA sequences. AB - DNA sequences of an 847 bp fragment of mitochondrial cytochrome oxidase subunit I (COI) gene and a 514 bp fragment of 16s rRNA gene were determined to examine the phylogenetic relationships of 12 Penaeoidea shrimp species (Penaeus chinensis, Penaeus japonicus, Penaeus penicillatus, Penaeus vannamei, Penaeus canaliculatus, Trachypenaeus curvirostris, Metapenaeus affinis, Metapenaeus ensis, Metapenaeopsis barbata, Parapenaeus fissuroides, Parapenaeopsis hardiwickii, Solenocera crassicomis). Both fragments of the swimming crab Portunus trituberculaus chosen as the outgroup were also sequenced. Intraspecific sequence divergence of 0.24-1.2% in the COI gene was found in 5 species, while no intraspecific variation was observed in the 16s rRNA gene. Three phylogenetic trees based on the 1361 bp combined sequences of COI and 16s rRNA were concordant in indicating the following suggestions: (1) phylogenetic relationship of the 11 Penaeidae species based on our result support the opinion of Burkenroad (Burkenroad, M.D. (1983). Crustacean Issues 3:279-290) on the basis of morphological features; (2) it seems more reasonable to class Solenocera crassicorni in the family Penaeidae; (3) the fragment of the COI gene chosen here appears to be a good marker for speciation studies and population analysis in Crustaceans, while the 16s rRNA gene fragment here seems suitable for examining phylogenetic relationships at the species or genus levels in Crustaceans. Our time estimates suggest that Penaeus and Metapenaeus might have separated about 6.38 x 10(6)-7.98 x 10(6) years BP in the post-Miocene, and the species separation within Metapenaeus and Penaeus might occur 0.08 x 10(6)-0.4 x 10(6) years BP in the late Pleistocene. PMID- 15524312 TI - Quantitative trait locus analysis of plasma cholesterol and triglyceride levels in C57BL/6J x RR F2 mice. AB - A highly significant cholesterol quantitative trait locus (QTL) (Cq6) was identified on chromosome 1 in C57BL/6J x RR F2 mice. The Cq6 was located over the gene for apolipoprotein A-Il (Apoa2), and the RR allele was associated with increased plasma cholesterol. C57BL/6J has Apoa2a alleles and RR has Apoa2b alleles. Three different Apoa2 alleles are known on the basis of amino acid substitutions at four residues. Analysis with partial Apoa2 congenic strains possessing Apoa2a, Apoa2b, and Apoa2C alleles revealed that the Apoa2b allele is unique in the ability to increase cholesterol among the three Apoa2 alleles, and that the Ala-to-Val substitution at residue 61 may be crucial as far as cholesterol metabolism is concerned. We also investigated the question of whether the Apoa1 gene is responsible for the cholesterol QTLs (Cq4 and Cq5) that had been identified previously on chromosome 9 in C57BL/6J x KK-Ay/a F2 and in KK x RR F2, but not in C57BL/6J x RR F2 mice. Similar to Apoa2 alleles, three different Apoal alleles with two successive amino acid substitutions were revealed among the strains. However, we could not correlate Apoal polymorphisms with the occurrence of QTLs in these three sets of F2 mice. PMID- 15524313 TI - Cloning and sequencing of the growth hormone gene of large yellow croaker and its phylogenetic significance. AB - Using conserved primers and the PCR reaction, the growth hormone (GH) gene and the 3'-UTR of the large yellow croaker (Pseudosciaena crocea) were amplified and sequenced. The gene structure was analyzed and compared to the GH genes of 5 other percoid fish downloaded from Genbank. Also the GH gene of the large yellow croaker and the genes from 14 Percoidei and 2 Labroidei species were aligned using Clustal X. A matrix of 564 bp was used to construct the phylogenetic tree using maximum parsimony and neighbor-joining methods. Phylogenetic trees by the two methods are identical in most of the clades with high bootstrap support. The results are also identical to those from morphological data. In general, this analysis does not support the monophyll of the families Centropomidae and Carangidae. But our GH gene tree indicates that the representative species of the families Sparidae and Sciaenidae are a monophyletic group. PMID- 15524314 TI - Identification of three novel insertion/deletion mutations in Wilson disease's gene. PMID- 15524315 TI - Polymorphisms of the second exon of MHC-DRB gene in Chinese local sheep and goat. PMID- 15524316 TI - Insulin resistance and abomasal motility disorders in cows detected by use of abomasoduodenal electromyography after surgical correction of left displaced abomasum. AB - OBJECTIVE: To determine the correlation between insulin concentrations and myoelectrical activity of the abomasum in cows with a left displaced abomasum (LDA). ANIMALS: 14 dairy cows with an LDA at the onset of lactation. PROCEDURE: During surgical correction of an LDA, 3 pairs of electrodes were placed in the smooth muscle of the gastrointestinal tract (abomasal body, pars pylorica, and duodenum) of each cow. Electromyographic recordings were obtained once per day for 7 days. Samples were collected and tested to determine concentrations of insulin, glucagon, cortisol, glucose, beta-hydroxybutyrate, and nonesterified fatty acids. RESULTS: All 14 cattle had high glucose and insulin concentrations at the time of admission, independent of ketosis. Concentrations of glucose and insulin decreased slowly after surgical treatment and were associated with a progressive increase in abomasoduodenal myoelectric activity. The 14 cows were allocated into 2 groups (suspected insulin-resistant cattle, n = 7; suspected non insulin-resistant cattle, 7) on the basis of persistent hyperinsulinemia during the postoperative period. Seven days after surgery, the abomasoduodenal myoelectric patterns were still significantly lower for the insulin-resistant cows, compared with patterns for the non-insulin-resistant cows. CONCLUSIONS AND CLINICAL RELEVANCE: Insulin resistance appears to be common in cows with an LDA. Analysis of results of this study reveals that abomasal atony in cows with an LDA depends on persistence of high serum concentrations of insulin. Results of this study could provide an explanation for a pathogenetic factor of LDAs and the frequent relapses of cattle affected by this condition. PMID- 15524317 TI - Effects of intracameral injection of preservative-free lidocaine on the anterior segment of the eyes in dogs. AB - OBJECTIVE: To evaluate effects of intracameral injection of preservative-free 1% and 2% lidocaine hydrochloride solution on the anterior segment of the eyes in dogs. ANIMALS: 16 adult healthy dogs (8 male and 8 female) judged to be free of ocular disease. PROCEDURE: Dogs were randomly assigned to 2 groups of 8 dogs each. Group 1 dogs received an intracameral injection of 0.10 mL of preservative free 1% lidocaine solution in the designated eye, and group 2 dogs received 0.10 mL of preservative-free 2% lidocaine solution in the designated eye. After injection, intraocular pressure was measured every 12 hours for 48 hours and then every 24 hours until 168 hours after injection. Slit-lamp biomicroscopy was performed preceding intracameral injection, 8 hours after injection, and then every 24 hours until 168 hours after injection. Ultrasonic pachymetry and specular microscopy were performed preceding intracameral injection and 72 and 168 hours after injection. Corneal thickness and endothelial cell density and morphology were compared with baseline measurements. RESULTS: No significant differences were found in intraocular pressure, corneal thickness, endothelial cell density, and morphologic features in either group, compared with baseline. A significant difference in aqueous flare was found for treated and control eyes 8, 24, and 48 hours after injection, compared with baseline. No significant difference in aqueous flare was found between treated and control eyes within either group. CONCLUSIONS AND CLINICAL RELEVANCE: No adverse ocular effects were detected after intracameral injection of preservative-free 1% or 2% lidocaine solution; thus, its use would be safe for intraocular pain management in dogs. PMID- 15524318 TI - Evaluation of antibody response and nonspecific lymphocyte blastogenesis following inoculation of a live attenuated bluetongue virus vaccine in goats. AB - OBJECTIVE: To evaluate vaccine safety, antibody response, and nonspecific lymphocyte blastogenesis following inoculation of a commercial monovalent live attenuated bluetongue virus (BTV) serotype 2 vaccine in goats. ANIMALS: 12 nonpregnant and nonlactating Saanen goats. PROCEDURE: 6 goats were inoculated with the monovalent live attenuated BTV serotype 2 vaccine, which has been widely used in Italy during the proceding 2 years. The other 6 goats were unvaccinated and represented negative controls. Nonspecific lymphocyte blastogenesis was evaluated 14 and 7 days before and 7, 21, and 49 days after vaccination by measuring DNA synthesis in peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin, concanavalin-A, and pokeweed mitogen. On the same days as lymphocyte blastogenesis, blood samples were taken to determine serum concentrations of anti-BTV antibodies. RESULTS: During the 7 weeks following vaccination, PBMCs obtained from vaccinated goats had a significantly decreased response to mitogens in terms of DNA synthesis, compared with PBMCs from the same goats before vaccination. Conversely during the experiment, no significant change was found in the response of the PBMCs obtained from unvaccinated goats. Starting from 21 days after vaccination, serum from vaccinated goats had anti-BTV antibodies. No anti-BTV antibodies were detected in the serum from unvaccinated goats. CONCLUSIONS AND CLINICAL RELEVANCE: Inoculation of goats with the monovalent live attenuated BTV serotype 2 vaccine described herein resulted in a profound depression of nonspecific lymphocyte blastogenesis, which might compromise the resistance of vaccinated goats to pathogens. PMID- 15524319 TI - Tissue Doppler assessment of diastolic and systolic alterations of radial and longitudinal left ventricular motions in Golden Retrievers during the preclinical phase of cardiomyopathy associated with muscular dystrophy. AB - OBJECTIVE: To quantify radial and longitudinal left ventricular free wall (LVFW) velocities in dogs during the preclinical phase of Golden Retriever muscular dystrophy (GRMD)-associated cardiomyopathy by use of tissue Doppler imaging (TDI). ANIMALS: 9 dogs with GRMD and 6 healthy control dogs. PROCEDURE: All dogs (< 3 years old) were examined via conventional echocardiography and 2-dimensional color TDI. Myocardial velocities in the LVFW were recorded from right parasternal ventricular short-axis (radial motion) and left apical 4-chamber (longitudinal motion) views. Cardiac assessments via TDI included maximal systolic and early and late diastolic LVFW velocities in the endocardial and epicardial layers (for radial motion) and in the basal and apical segments (for longitudinal motion) (for longitudinal motion), RESULTS: -No notable ventricular dilatation or alteration of inotropism was detected in dogs with GRMD via conventional echocardiography. Compared with healthy dogs, endocardial velocities were significantly decreased in dogs with GRMD, resulting in marked decreases in radial myocardial velocity gradients during systole and early and late diastole. Similarly, basal and apical velocities were significantly decreased in systole and the former also in early diastole, resulting in significant decreases in the 2 corresponding longitudinal myocardial velocity gradients. The radial epicardial and longitudinal late diastolic velocities were comparable in the 2 groups. CONCLUSION AND CLINICAL RELEVANCE: Results indicated that GRMD-associated cardiomyopathy in dogs is associated with early marked dysfunction of both radial and longitudinal LVFW motions. These combined regional myocardial abnormalities might be useful criteria for detection of dilated cardiomyopathy at the preclinical stage of the disease in dogs. PMID- 15524320 TI - Effects of alpha2-adrenergic receptor agonists on urine production in horses deprived of food and water. AB - OBJECTIVE: To quantitate the dose- and time-related effects of IV administration of xylazine and detomidine on urine characteristics in horses deprived of feed and water. ANIMALS: 6 horses. PROCEDURE: Feed and water were withheld for 24 hours followed by i.v. administration of saline (0.9% NaCI) solution, xylazine (0.5 or 1.0 mg/kg), or detomidine (0.03 mg/kg). Horses were treated 4 times, each time with a different protocol. Following treatment, urine and blood samples were obtained at 15, 30, 60, 120, and 180 minutes. Blood samples were analyzed for PCV and serum concentrations of total plasma solids, sodium, and potassium. Urine samples were analyzed for pH and concentrations of glucose, proteins, sodium, and potassium. RESULTS: Baseline (before treatment) urine flow was 0.30 +/- 0.03 mL/kg/h and did not significantly change after treatment with saline solution and low-dose xylazine but transiently increased by 1 hour after treatment with high dose xylazine or detomidine. Total urine output at 2 hours following treatment was 312 +/- 101 mL versus 4,845 +/- 272 mL for saline solution and detomidine, respectively. Absolute values of urine concentrations of sodium and potassium also variably increased following xylazine and detomidine administration. CONCLUSIONS AND CLINICAL RELEVANCE: Xylazine and detomidine administration in horses deprived of feed and water causes transient increases in urine volume and loss of sodium and potassium. Increase in urine flow is directly related to dose and type of alpha2-adrenergic receptor agonist. Dehydration in horses may be exacerbated by concurrent administration of alpha2-adrenergic receptor agonists. PMID- 15524321 TI - Evaluation of hepatosplanchnic circulation and intestinal oxygenation in dogs with a condition that mimicked septic shock induced by continuous infusion of a low dose of lipopolysaccharide. AB - OBJECTIVE: To determine whether continuous infusion of a low dose of lipopolysaccharide (LPS) to induce a condition mimicking septic shock in dogs would affect systemic and hepatosplanchnic circulation and oxygenation. ANIMALS: 12 healthy adult Beagles. PROCEDURE: Dogs received a low dose of LPS (Escherichia coli O55:B5) by continuous IV infusion at a rate of 1 microg/kg/h for 8 hours. Systemic hemodynamics; systemic oxygenation; blood flow in the cranial mesenteric artery, common hepatic artery, and portal vein; intestinal and hepatic tissue blood flow; mesenteric oxygenation; and intramucosal Pco2 were examined before and at selected time points after onset of the LPS infusion. RESULTS: After onset of the LPS infusion, cardiac index increased and mean arterial pressure (MAP) and systemic vascular resistance decreased, which is characteristic of the hyperdynamic state in septic patients. Hepatosplanchnic blood flow increased during the hyperdynamic state. Intestinal Pco2 was increased even when blood flows increased. During the latter half of the experimental period, MAP was maintained but hepatosplanchnic blood flows decreased and intestinal Pco2 increased further. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of the results suggested that hepatosplanchnic blood flow enters the hyperdynamic state during the early stages of sepsis and that intestinal tissue oxygenation is threatened even when hepatosplanchnic blood flow is increased or maintained. Hence, improvement of hepatosplanchnic circulation and intestinal tissue oxygenation is important in dogs with clinical evidence of a septic condition. PMID- 15524323 TI - Distribution of Escherichia coli O157:H7 within and among cattle operations in pasture-based agricultural areas. AB - OBJECTIVE: To determine the distribution of Escherichia coli O157:H7 in pasture based cattle production areas. SAMPLE POPULATION: Two 100-km2 agricultural areas consisting of 207 pasture, 14 beef-confinement, and 3 dairy locations within 24 cattle operations. PROCEDURE: 13,726 samples from cattle, wildlife, and water sources were obtained during an 11-month period. Escherichia coli O157:H7 was identified by use of culture and polymerase chain reaction assays and characterized by pulsed-field gel electrophoresis (PFGE). RESULTS: Odds of recovering E coli O157:H7 from feeder-aged cattle were > 4 times the odds for cow calf or dairy cattle. There was no difference in prevalence for pastured versus confined cattle after controlling for production age group. Number of samples collected (37 to 4,829), samples that yielded E coli O157:H7 (0 to 53), and PFGE subtypes (0 to 48) for each operation varied and were highly correlated. Although most PFGE subtypes were only detected once, 17 subtypes were detected on more than 1 operation. Ten of 12 operations at which E coli O157:H7 was detected had at least 1 subtype that also was detected on another operation. We did not detect differences in the probability of having the same subtype for adjacent operations, nonadjacent operations in the same study area, or operations in the other study area. CONCLUSIONS AND CLINICAL RELEVANCE: Strategies aimed at controlling E coli O157:H7 and specific subtypes should account for the widespread distribution and higher prevalence in feeder-aged cattle regardless of production environment and the fact that adjacent and distant cattle operations can have similar subtypes. PMID- 15524322 TI - Expression, bioactivity, and clinical assessment of recombinant feline erythropoietin. AB - OBJECTIVE: To determine the activity of recombinant feline erythropoietin (rfEPO) in murine bioassays and evaluate its efficacy and safety in cats with erythropoietin-dependent nonregenerative anemia. ANIMALS: 26 cats (group 1, 19 cats with anemia attributed to chronic kidney disease [CKD]; group 2, 7 cats with CKD and recombinant human erythropoietin [rhEPO]-induced red cell aplasia [RCA]). PROCEDURE: The rfEPO was synthesized by use of Chinese hamster ovary (CHO) cells transfected with feline erythropoietin complementary DNA. Preclinical assessments of rfEPO included an erythroid cell proliferation assay and measurements of reticulocytosis in Balb/C mice. Clinical assessments of cats included hematologic, biochemical, and clinical examinations during 12 (group 1) or 6 (group 2) months of rfEPO treatment. RESULTS: Biological activity of rfEPO was broadly equivalent to rhEPO in preclinical murine bioassays. Median Hct and absolute reticulocyte count in cats increased significantly during the first 3 weeks of rfEPO treatment, and median Hct generally could be maintained within a target range of 30% to 40% with periodic adjustments of rfEPO doses. Unexpectedly, 5 cats in group 1 and 3 cats in group 2 that initially responded to rfEPO treatment again developed anemia that was refractory to additional rfEPO treatments, even at higher doses. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with rfEPO can reestablish active erythropoiesis in most cats with CKD, even those with anemia attributable to rhEPO-induced RCA. Unfortunately, development of RCA during treatment with CHO cell-derived recombinant erythropoietin proteins was not eliminated as a serious safety concern, even for this feline-specific preparation. PMID- 15524324 TI - Effects of ischemia and the cyclooxygenase inhibitor flunixin on in vitro passage of lipopolysaccharide across equine jejunum. AB - OBJECTIVE: To determine whether ischemia and flunixin affect in vitro lipopolysaccharide (LPS) absorption in samples of the jejunum of horses. ANIMALS: 12 horses. PROCEDURE: Horses were anesthetized, a midline celiotomy was performed, and the jejunum was located. Two 30-cm sections of jejunum (60 cm apart) were selected. One segment was designated as control tissue; ischemia was induced in the other segment for 120 minutes. Horses were then euthanatized. Mucosa from each jejunal segment was mounted on Ussing chambers and treated with or without flunixin. Tissues from 6 horses were used to assess permeability to radiolabeled LPS; mucosal samples from the remaining 6 horses were incubated with fluorescent-labeled LPS (FITC-LPS) and examined histologically. Production of tumor necrosis factor-alpha (TNF-alpha) and production of LPS-binding protein (LBP) were assessed as indicators of mucosal response to LPS. RESULTS: Ischemia significantly increased mucosal permeability to LPS, but by 180 minutes, the mucosa was not more permeable than control tissue. Flunixin treatment adversely affected intestinal barrier function throughout the experiment but did not result in increased mucosal permeability to LPS. Compared with control tissues, LBP production was increased by ischemia and reduced by exposure to LPS. In ischemic tissue, FITC-LPS entered the lamina propria but TNF-alpha was produced on the mucosal side only, indicating little response to the absorbed LPS. CONCLUSIONS AND CLINICAL RELEVANCE: Ischemia increased LPS passage across equine jejunal mucosa. Flunixin delayed mucosal recovery but did not exacerbate LPS absorption. Evaluation of the clinical importance of flunixin-associated delayed mucosal recovery requires further in vivo investigation. PMID- 15524326 TI - Effect of a commercial anion dietary supplement on acid-base balance, urine volume, and urinary ion excretion in male goats fed oat or grass hay diets. AB - OBJECTIVE: To determine whether feeding a commercial anionic dietary supplement as a urinary acidifier to male goats may be useful for management of urolithiasis. ANIMALS: 8 adult sexually intact male Toggenburg, Saanen, and Nubian goats. PROCEDURE: Goats were randomly assigned by age-, breed-, and weight matched pairs to an oat or grass hay diet that was fed for 12 days. On days 13 to 14 (early sample collection time before supplementation), measurements were made of blood and urine sodium, potassium, calcium, magnesium, chloride, phosphorus, and sulfur concentrations; blood and urine pH; urine production; and water consumption. During the next 28 days, the anionic dietary supplement was added to the oat and grass hay diets to achieve a dietary cation-anion difference of 0 mEq/100g of dry matter. Blood and urine samples were analyzed during dietary supplementation on days 12 to 13 (middle sample collection time) and 27 to 28 (late sample collection time). RESULTS: Blood bicarbonate, pH, and urine pH of goats fed grass hay and goats fed oat hay were significantly decreased during the middle and late sample collection times, compared with the early sample collection time. Water consumption and urine production in all goats increased significantly during the late sample collection time, compared with the early sample collection time. CONCLUSIONS AND CLINICAL RELEVANCE: The anionic dietary supplement used in our study increases urine volume, alters urine ion concentrations, and is an efficacious urinary acidifier in goats. Goats treated with prolonged anionic dietary supplementation should be monitored for secondary osteoporosis from chronic urinary calcium loss. PMID- 15524325 TI - Effect of meloxicam and carprofen on renal function when administered to healthy dogs prior to anesthesia and painful stimulation. AB - OBJECTIVE: To determine whether administration of the nonsteroidal anti inflammatory drugs meloxicam or carprofen to healthy dogs that were subsequently anesthetized and subjected to painful electrical stimulation has adverse effects on renal function as measured by glomerular filtration rate (GFR) and evaluation of serum concentrations of urea and creatinine. ANIMALS: 6 male and 6 female healthy young-adult Beagles. PROCEDURE: A study was conducted in accordance with a randomized crossover Latin-square design. One of 3 treatments (saline [0.9% NaCl] solution, 0.2 mg of meloxicam/kg, or 4.0 mg of carprofen/kg) was administered i.v. 1 hour before anesthesia was induced by use of drugs in accordance with a standard anesthetic protocol (butorphanol tartrate and acepromazine maleate as preanesthetic medications, ketamine hydrochloride and diazepam for induction, and maintenance with isoflurane). Anesthetized dogs were subjected to intermittent electrical stimulation for 30 minutes. Direct, mean arterial blood pressure; heart rate; and respiratory rate were monitored. End tidal isoflurane concentration was maintained at 1.5 times the minimum alveolar concentration. The GFR, as measured by plasma clearance of 99mTc diethylenetriaminepentaacetic acid, and serum concentrations of serum and creatinine were determined 24 hours after induction of anesthesia. RESULTS: Neither meloxicam nor carprofen significantly affected GFR or serum concentrations of urea and creatinine, compared with values for the saline treatment. CONCLUSIONS AND CLINICAL RELEVANCE: When administered 1 hour before onset of anesthesia and painful electrical stimulation, meloxicam or carprofen did not cause clinically important alterations of renal function in young healthy dogs. PMID- 15524327 TI - Evaluation of environmental and management-related risk factors associated with chronic mastitis in sows. AB - OBJECTIVE: To evaluate environmental and management-related risk factors associated with chronic mastitis in sows. ANIMALS: 1,254 sows from 76 herds. PROCEDURE: Prevalence of chronic mastitis was determined by a veterinarian who performed clinical examinations at the time of weaning and approximately 1 week later in a sample of the sow population on each farm. Information concerning environmental factors and management practices was collected. In addition, the herd veterinarian made an assessment of the farmer's skills in swine production. RESULTS: Use of partly slatted floors in the farrowing pens, use of disinfectants between batches in the farrowing and breeding areas, feeding lactating sows whey, and avoiding cutting or grinding of the piglets' teeth were significantly associated with a decreased risk of chronic mastitis. A high hygienic standard on the farm, as determined by the herd veterinarian, was associated with a significant reduction in the prevalence of mastitis. CONCLUSIONS AND CLINICAL RELEVANCE: Chronic mastitis in sows is a common disease that has a negative influence on productivity. Results indicate that certain management practices and environmental factors influenced the development of mastitis, which may contribute to the development of methods useful for controlling the disease. PMID- 15524328 TI - Evaluation of vaccination with Neospora caninum protein for prevention of fetal loss associated with experimentally induced neosporosis in sheep. AB - OBJECTIVE: To evaluate the immunologic response of a killed tachyzoite vaccine against Neospora caninum and its effectiveness in preventing fetal loss associated with experimentally induced neosporosis in sheep. ANIMALS: 30 Dorset ewes. PROCEDURE: Ewes were randomly allocated to receive vaccination on days 1 and 60 of the study with a killed N caninum tachyzoite preparation in a commercially available adjuvant or a saline-adjuvant mixture. A ram was placed on pasture with the ewes from days 15 to 60. Blood was collected from ewes before primary and booster vaccinations and prior to experimental challenge with N caninum tachyzoite performed on day 90; sera were assessed via Neospora agglutination (NA) and immunofluorescence antibody (IFA) assays. Blood was collected from lambs before they suckled, and sera were tested for antibodies against N caninum. RESULTS: Of the 14 vaccinated ewes that became pregnant, 12 gave birth to live-born lambs; in contrast, 5 of 11 pregnant control ewes gave birth to live-born lambs. Whereas vaccination improved fetal survival in pregnant ewes challenged with N caninum tachyzoites, it did not appear to have any appreciable effect on transmission of N caninum to offspring, as indicated by results of NA and IFA assays. CONCLUSIONS AND CLINICAL RELEVANCE: The N caninum tachyzoite vaccine used in this study appeared to provide protection against fetal loss associated with experimentally induced neosporosis in a high proportion of pregnant ewes. PMID- 15524329 TI - Use of computed tomography angiography to evaluate the vascular anatomy of the distal portion of the forelimb of horses. AB - OBJECTIVE: To provide a detailed description of the vascular anatomy of the distal portion of the forelimbs of horses by use of computed tomography angiography (CTA). SAMPLE POPULATION: 6 forelimbs of 5 horses and 1 forelimb from an equine cadaver; none of the horses had orthopedic or vascular disease. PROCEDURE: Horses were anesthetized and CTA was conducted on the dependent forelimb. A catheter was inserted in the median artery, and contrast medium was infused at a rate of 3 mL/s. A computed tomography (CT) scanner was used to obtain contiguous slices from the region of the proximal sesamoid bones to the toe. All horses were allowed to recover from anesthesia. To help identify vessel patterns in the distal portion of the forelimb, the median artery and lateral palmar digital vein of a heparinized forelimb obtained from an equine cadaver were infused with red and blue polymethylmethacrylate and the distal portion of that forelimb was then sectioned to correspond to CTA images. RESULTS: Vessel patterns in CTA images matched vascular anatomic structures of the cadaver forelimb and were consistent with published anatomic structures. Major and minor vessels were consistently visible in CTA images of all horses. There were no complications reported in any horses. CONCLUSIONS AND CLINICAL RELEVANCE: Use of CTA provided a highly detailed depiction of the vasculature of the distal portion of the equine forelimb. This was a safe technique and should be useful in the evaluation of the blood supply to the distal portion of the forelimb. PMID- 15524330 TI - Effect of proinflammatory mediators and glucocorticoids on L-selectin expression in peripheral blood neutrophils from dairy cows in various stages of lactation. AB - OBJECTIVE: To determine whether proinflammatory mediators and glucocorticoids affect CD62L(L-selectin) expression on peripheral blood neutrophils from cows in various stages of lactation. ANIMALS: 100 healthy dairy cows during early (13.1 +/- 0.79 days after parturition; n = 31), peak (58.7 +/- 1.64 days after parturition; 31), and mid (137.2 +/- 2.59 days after parturition; 38) lactation. PROCEDURE: In vitro effects of relevant proinflammatory mediators that are released in response to mastitis caused by gram-negative bacteria such as lipopolysaccharide (endotoxin), tumor necrosis factor-alpha, and platelet activating factor (PAF) on CD62L expression on bovine neutrophils were assessed by flow cytometry. Influences of cortisol and dexamethasone on CD62L expression on bovine neutrophils were also investigated. RESULTS: Basal CD62L expression on neutrophils from cows during early, peak, and mid lactation were similar. Lipopolysaccharide and tumor necrosis factor-alpha had no effect on CD62L expression on neutrophils from cows at any stage of lactation. Conversely, PAF elicited a time- and dose-dependent, down regulatory effect on CD62L expression. However, no differential shedding of CD62L from neutrophils of cows at any stage of lactation were detected. In addition, no effects on CD62L expression on bovine neutrophils after whole blood incubation with cortisol or dexamethasone were observed. Incubation with glucocorticoids did not prevent the down regulatory effect of PAF on CD62L expression. CONCLUSIONS AND CLINICAL RELEVANCE: Comparable basal CD62L expression on bovine neutrophils and equal amounts of CD62L shedding from bovine neutrophils during all stages of lactation suggest that variations in CD62L density are not a likely cause of susceptibility of cows to coliform induced mastitis during early lactation. PMID- 15524331 TI - Evaluation of the immunogenicity of dietary proteins in cats and the influence of the canning process. AB - OBJECTIVE: To characterize the antigen-specific immune response to dietary proteins in cats and evaluate whether there was a qualitative or quantitative difference between the responses to dietary proteins when those proteins were fed unprocessed or as part of a canned diet. ANIMALS: 14 healthy domestic shorthair cats. PROCEDURE: Cats were fed 2 dietary proteins (soy and casein) either as unprocessed aqueous suspensions or as part of canned diets for 21 days. Serum IgG and IgA and salivary IgA were assayed by indirect ELISA, and antigen-specific proliferation of mesenteric lymph node-derived lymphocytes was determined. RESULTS: Robust serum IgG and IgA responses to dietary proteins were elicited, irrespective of the form in which they were fed. Salivary IgA responses to unprocessed proteins were not detected. However, a significant salivary IgA response to the protein isolated from the canned casein diet was observed in cats fed canned casein but not in those fed unprocessed casein. Lymphocyte proliferation to the antigens was slight, and there were no significant differences between groups. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that cats develop robust serum IgG and IgA responses to dietary proteins when fed as either aqueous suspensions or as part of canned diets. For certain proteins, there may be an increase and a qualitative difference in the immunogenicity of canned diets, compared with unprocessed proteins. Canned diets may not be ideal for management of cats with enteritis. PMID- 15524332 TI - Tumor necrosis factor-alpha, interleukin-6, serum amyloid A, haptoglobin, and cortisol concentrations in sows following intramammary inoculation of Escherichia coli. AB - OBJECTIVE: To determine whether concentrations of proinflammatory cytokines, acute-phase proteins, and cortisol differ at parturition among 3 categories of sows (noninoculated, clinically affected and nonaffected following intramammary inoculation with Escherichia coll). ANIMALS: 16 sows. PROCEDURE: Sows were allocated to inoculated (n = 12) or noninoculated (4) groups. Inoculated sows received intramammary administration of E coli (serotype O127) during the 24-hour period preceding parturition. Blood samples were collected from noninoculated and inoculated sows for 3 consecutive days within 3 to 11 days before farrowing and inoculation. Samples were also collected 0, 24, 48, 72, and 96 hours after farrowing and inoculation. Inoculated sows were further categorized as affected (4 sows) or nonaffected (8 sows) based on clinical signs of disease. Serum tumor necrosis factor (TNF)-alpha, plasma interleukin (IL)-6, and serum amyloid A (SAA) concentrations were measured by use of ELISA; serum haptoglobin concentration was assayed by use of a hemoglobin-binding method; and plasma cortisol concentration was determined by use of radioimmunoassay. RESULTS: Plasma or serum concentrations of TNF-alpha, IL-6, and SAA of both categories of inoculated sows were significantly increased by 24 hours after intramammary inoculation of E coli, compared with concentrations in noninoculated sows. Concentrations of serum TNF-alpha and plasma IL-6 were significantly higher in inoculated sows that developed clinical mastitis than in nonaffected inoculated sows. CONCLUSIONS AND CLINICAL RELEVANCE: Concentrations of TNF-alpha and IL-6 are promising markers for the identification of periparturient sows with subclinical coliform mastitis. Identification of such sows should help improve the health and survival of piglets. PMID- 15524333 TI - Comparison of inhibitory effects of glucosamine and mannosamine on bovine articular cartilage degradation in vitro. AB - OBJECTIVE: To compare the inhibitory effects of glucosamine and mannosamine on articular cartilage degradation and the effects on chondrocyte viability in vitro. SAMPLE POPULATION: Bovine articular cartilage explants. PROCEDURES: Explants were cultured in commercial medium for 48 hours. Cartilage was exposed to medium containing 10% fetal bovine serum, 10 microg of lipopolysaccharide/mL, and 0.5, 1.0, 2.5, 5.0, and 10.0 mg of glucosamine or mannosamine/mL for 24 hours. Nitric oxide (NO) production (nitrite concentration) and proteoglycan (PG) release (PG concentration) in media were measured. Cartilage extracts were analyzed via zymography to detect gelatinolytic activity. At the end of the experiment, explants were assessed for chondrocyte viability. RESULTS: Addition of lipopolysaccharide resulted in increased NO production and PG release, but no increase in gelatinolytic activity, compared with controls. Glucosamine and mannosamine at concentrations as low as 0.5 mg/mL inhibited NO production. Glucosamine inhibited PG release at a minimum concentration of 1.0 mg/mL, whereas mannosamine inhibited PG release at a concentration of 0.5 mg/mL. Concentrations of glucosamine < or = 5.0 mg/mL did not adversely affect chondrocyte viability; however, at a concentration of 10.0 mg/mL, cell death was evident. Mannosamine had a toxic effect at a concentration of 5.0 mg/mL and was associated with pronounced chondrocyte death at a concentration of 10.0 mg/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Glucosamine and mannosamine inhibit selected indices of bovine articular cartilage degradation at concentrations that do not affect chondrocyte viability. The potential for cytotoxic effects at higher concentrations underscores the importance of establishing appropriate dosage regimens for these aminomonosaccharides. PMID- 15524334 TI - Evaluation of a rapid single multiplex microsatellite-based assay for use in forensic genetic investigations in dogs. AB - OBJECTIVE: To develop a set of microsatellite markers, composed of a minimal number of these markers, suitable for use in forensic genetic investigations in dogs. SAMPLE POPULATION: Blood, tissue, or buccal epithelial cells from 364 dogs of 85 breeds and mixed breeds and 19 animals from related species in the family Canidae. PROCEDURE: 61 tetranucleotide microsatellite markers were characterized on the basis of number and size of alleles, ease of genotyping, chromosomal location, and ability to be coamplified. The range in allele size, number of alleles, total heterozygosity, and fixation index for each marker were determined by use of genotype data from 383 dogs and related species. Polymorphism information content was calculated for several breeds of dogs. RESULTS: 7 microsatellite markers could be coamplified. These markers were labeled with fluorescent dyes, multiplexed into a single reaction, and optimized for resolution in a commercial genetic analyzer. The multiplex set was used to identify sires for 2 mixed litters. The test was not species specific; genotype information collected for wolves, coyotes, jackals, New Guinea singing dogs, and an African wild dog could not distinguish between these species. CONCLUSIONS AND CLINICAL RELEVANCE: This set of 7 microsatellite markers is useful in forensic applications (ie, identification of dogs and determination of parentage) in closely related animals and is applicable to a wide range of species belonging to the family Canidae. PMID- 15524335 TI - Combating mortality from severe sepsis. PMID- 15524336 TI - Artificial oxygen carriers for trauma: myth or reality. AB - Two-unit blood transfusion accounts for one third of blood use in trauma patients. Artificial oxygen carriers have no need for cross-match, allow volume expansion, prolonged storage at 1-38 degrees C, and improve rheology to reverse ischaemia. Haemoglobin-based oxygen carriers can provide a vital alternative given the predicted future shortfall in blood donations. PMID- 15524337 TI - Radial artery as a conduit for coronary artery bypass graft. AB - Since its resurgence in the 1990s, some 30 years after its abandonment as an alternative conduit, the radial artery has rapidly gained popularity. This article describes the application of this conduit in coronary grafting and details the controversies surrounding its use. A historical account as well as technical aspects such as operative technique are also discussed. PMID- 15524338 TI - Pharmacogenetics and cardiovascular disease management. AB - Patients differ in their response to drugs. Part of this variability may reflect genetically-determined characteristics of target genes or metabolizing enzymes. A knowledge of an individual's genetic makeup could allow drug therapy to be targeted at those most likely to benefit. PMID- 15524339 TI - Older drivers and motor vehicle crashes. AB - Older drivers are a source of great public concern, particularly after well publicized incidents in which an older driver has caused a grievous event. As their numbers grow, understanding the risks that older drivers pose to themselves and other road users is important for appropriate clinical and policy decisions. PMID- 15524340 TI - Radiotherapy for prostate cancer and sexual functioning. AB - The incidence of erectile dysfunction after radiotherapy for prostate cancer is high. A multifactorial aetiology has to be considered, taking into account pretreatment erectile function. Patients need to be informed about effective treatments such as sildenafil and intracavernosal injections. PMID- 15524341 TI - Fatty liver. AB - Fatty liver is increasingly becoming more problematic from a clinical perspective. This article describes fatty liver and its clinical presentation. The current understanding of processes underlying fatty liver is reviewed, as well as the evidence for therapeutic options. PMID- 15524342 TI - Breaking bad news: practical advice for busy doctors. AB - Breaking bad news is a difficult task faced daily by the busy doctor. This article draws on some of the literature on the topic and offers some practical advice on how to break bad news. PMID- 15524343 TI - Acute management of paediatric respiratory failure. PMID- 15524344 TI - Flexible (part-time) preregistration house officer training: review of Thames and London deanery experiences. AB - Flexible training has been in greatest demand at the specialist registrar grade, and to a lesser extent in the senior house officer grade. This article describes the experiences of a much smaller group, flexible preregistration house officers. PMID- 15524345 TI - The first successful renal graft in man. PMID- 15524346 TI - Squamous cell carcinoma of the nasal vestibule presenting as chronic vestibulitis. PMID- 15524347 TI - Leiomyoma of the nasal cavity: case report and literature review. PMID- 15524349 TI - A diagnosis not to miss. PMID- 15524350 TI - Anaesthetist or intensivist: continued debate. PMID- 15524351 TI - Anaesthetist or intensivist: continued debate. PMID- 15524352 TI - Anaesthetist or intensivist: continued debate. PMID- 15524353 TI - Anaesthetist or intensivist: continued debate. PMID- 15524354 TI - Copying letters to patients. PMID- 15524355 TI - Swollen and cyanotic arm: a new cause? PMID- 15524356 TI - Looking after your family's future. PMID- 15524357 TI - Is preoperative pacing for bifascicular and trifascicular heart block necessary? PMID- 15524358 TI - Higher-order complex source for elegant Laguerre-Gaussian waves. AB - We introduce a higher-order complex source that generates elegant Laguerre Gaussian waves with radial mode number n and angular mode number m. We derive the integral and differential representations for the elegant Laguerre-Gaussian wave that in the appropriate limit yields the corresponding elegant Laguerre-Gaussian beam. From the spectral representation of the elegant Lauguerre-Gaussian wave we determine the first three orders of nonparaxial corrections for the corresponding paraxial elegant Laguerre-Gaussian beam. PMID- 15524359 TI - Direct visualization of surface-plasmon bandgaps in the diffuse background of metallic gratings. AB - When a surface plasmon propagates along a microrough grating, it interacts with the periodic plus the random roughness and emits light into the diffuse background, which can present intensity maxima called diffuse light bands. We reexamine previous studies on these bands within the framework of recent studies on photonic surfaces and show that the phenomenon of diffuse light provides an experimental technique for directly imaging the dispersion relation of surface plasmons, including the gap that, under appropriate circumstances, opens in the reciprocal grating space. PMID- 15524360 TI - Wideband tunable fiber short-pass filter based on side-polished fiber with dispersive polymer overlay. AB - It is known that dispersive fiber has an LP01 mode cutoff wavelength that acts as a wavelength filter. With a dispersive polysiloxane polymer overlay on a side polished single-mode fiber, an in-line tunable fiber short-pass filter with a tuning range of approximately 400 nm (1250-1650 nm) and a temperature variation of 15 degrees C was demonstrated. The rejection efficiency was greater than 50 dB, whereas the insertion and polarization-dependent losses were below 0.26 and 0.09 dB, respectively. PMID- 15524361 TI - Interrogating fiber Bragg grating sensors by thermally scanning a demultiplexer based on arrayed waveguide gratings. AB - We evaluate a wavelength interrogation technique based on an arrayed waveguide grating (AWG). Initial results show that the Bragg wavelength of fiber Bragg grating (FBG) sensors can be precisely interrogated by thermally scanning an AWG based demultiplexer. The technique potentially offers a low-cost, compact, and high-performance solution for the interrogation of FBG distributed sensors and multisensor arrays. PMID- 15524362 TI - Differential birefringence in Bragg gratings in multicore fiber under transverse stress. AB - We present experimental measurements of the peak splitting of the reflection spectra of fiber Bragg gratings as a result of birefringence induced by transverse loading of a multicore fiber. Measurements show that the splitting is a function of the applied load and the direction of the load relative to the azimuth of the fiber. A model for calculating the stress in the fiber that is due to an applied load is in good agreement with our experimental observations. PMID- 15524363 TI - Temporally and spectrally resolved imaging of laser-induced plasmas. AB - We report a hybrid imaging technique capable of performing measurements of the spatial, temporal, and spectral emission characteristics of laser-induced plasmas by use of a single detection system. We apply this technique to study the plasma produced by laser ablation of LiNbO3 and observe phenomena not seen in such detail with standard instruments. These include extreme line broadening up to a few nanometers accompanied by self-absorption near the target surface, and expansion dynamics that differ strongly between the different species. Overall, the wealth of quantitative information provided by this novel technique sheds new light on processes occurring during plasma expansion. PMID- 15524364 TI - High-precision measurements of light-induced torque on absorbing microspheres. AB - Laser beams have been demonstrated to be capable of exerting torque as well as forces on microparticles. Using a custom magneto-optic manipulator, we directly measured the torque exerted by laser light on absorbing microspheres as a result of the transfer of spin angular momentum. A general method for measuring torque has been developed, and the experimental apparatus has shown a sensitivity of approximately 1 pN/nm. PMID- 15524365 TI - Influence of the order of diattenuator, retarder, and polarizer in polar decomposition of Mueller matrices. AB - Polar decomposition consists of representing an arbitrary Mueller matrix with a product of three simpler matrices, but, since these matrices do not commute, the result depends on the order in which they are multiplied. We show that the six possible decompositions can be classified into two families and that one of these families always leads to physical elementary matrices, whereas the other does not. PMID- 15524366 TI - Electrochromic blueshift in polymer-dispersed liquid-crystal cells. AB - Electrochromic blueshift in the absorption band of polymer-dispersed liquid crystal cells is reported as a function of applied electric field. The changes in the peak absorption wavelength, absorption broadening, and their possible relationships with the nonlinear optical properties of polymer-dispersed liquid crystal cells are discussed. PMID- 15524367 TI - Use of ultrafast-laser-driven microexplosional for fabricating three-dimensional void-based diamond-lattice photonic crystals in a solid polymer material. AB - Micro-sized void spheres are successfully generated in a solid polymer by use of a tightly focused femtosecond laser beam from a high-repetition-rate laser oscillator. Confocal reflection images show that the void spheres are longitudinal rotational symmetric ellipsoids with a ratio of long to short axes of approximately 1.5. Layers of void spheres are then stacked to create three dimensional diamond-lattice photonic crystals. Three gaps are observed in the [100] direction with a suppression rate of the second gap of up to approximately 75% for a 32-layer structure. The observed first- and second-order gaps shift to longer and shorter wavelengths, respectively, as the angle of incidence increases. PMID- 15524368 TI - Loss-coupled distributed-feedback lasers with amplified optical feedback for optical microwave generation. AB - Multisection semiconductor lasers for optical microwave generation have been fabricated that consist of a loss-coupled distributed feedback (LC-DFB), a phase control, and an amplifier section. High-frequency self-pulsations are generated according to the concept of a single-mode laser with short optical feedback. The effect of the optical feedback via the phase control and the amplifier section on the self-pulsation is apparently shown as a result of the superior single-mode characteristic of the LC-DFB section. Continuous frequency tuning is achieved in the range of 17-35 GHz. PMID- 15524369 TI - Passively synchronized erbium (1550-nm) and ytterbium (1040-nm) mode-locked fiber lasers sharing a cavity. AB - Erbium and ytterbium fiber lasers were firmly synchronized by nonlinear interaction in active media placed in the same cavity. A two-color femtosecond picosecond pulse train at largely separate wavelengths of 1.55 and 1.04 microm was generated. Optimizing the laser cavity to enhance the cross-phase modulation in the gain materials has yielded a large mismatch of 20 microm between the two laser cavities. PMID- 15524370 TI - High-speed wide-field time-gated endoscopic fluorescence-lifetime imaging. AB - We report the development of a high-speed wide-field fluorescence-lifetime imaging (FLIM) system that provides fluorescence-lifetime images at rates of as many as 29 frames/s. A FLIM multiwell plate reader and a potentially portable FLIM endoscopic system operating at 355-nm excitation have been demonstrated. PMID- 15524371 TI - Polarization-resolved second-harmonic-generation optical coherence tomography in collagen. AB - We describe a novel imaging technique, second-harmonic-generation optical coherence tomography (SHOCT). This technique combines the spatial resolution and depth penetration of optical coherence tomography (OCT) with the molecular sensitivity of second-harmonic-generation spectroscopy. As a consequence of the coherent detection required for OCT, polarization-resolved images arise naturally. We demonstrate this new technique on a skin sample from the belly of Icelandic salmon, acquiring polarization-resolved SHOCT and OCT images simultaneously. PMID- 15524372 TI - Coherence-gated wave-front sensing in strongly scattering samples. AB - We show that, by coherence-gate rejection of out-of-focus light, wave-front distortions can be measured in the presence of a scattering background that is dominant by several orders of magnitude. Applications are expected for multiphoton and confocal laser-scanning microscopy. PMID- 15524373 TI - Simple time-domain optical method for estimating the depth and concentration of a fluorescent inclusion in a turbid medium. AB - A simple time-domain optical method for estimating the depth and concentration of fluorescent inclusions in turbid media is described. We demonstrate direct depth estimation of a localized fluorescent object from the temporal position of the temporal point-spread function maximum. The depth estimation permits recovery of the fluorophore concentration, both of which are essential quantities for optical molecular imaging studies. Since the maximum is independent of the fluorophore concentration, excitation laser power, detector gain, and other system-dependent factors, this method ensures a robust and efficient approach. PMID- 15524374 TI - Micromotor endoscope catheter for in vivo, ultrahigh-resolution optical coherence tomography. AB - A distally actuated, rotational-scanning micromotor endoscope catheter probe is demonstrated for ultrahigh-resolution in vivo endoscopic optical coherence tomography (OCT) imaging. The probe permits focus adjustment for visualization of tissue morphology at varying depths with improved transverse resolution compared with standard OCT imaging probes. The distal actuation avoids nonuniform scanning motion artifacts that are present with other probe designs and can permit a wider range of imaging speeds. Ultrahigh-resolution endoscopic imaging is demonstrated in a rabbit with <4-microm axial resolution by use of a femtosecond Cr:forsterite laser light source. The micromotor endoscope catheter probe promises to improve OCT imaging performance in future endoscopic imaging applications. PMID- 15524375 TI - Monolithically integrated bacteriorhodopsin-GaAs/GaAlAs phototransceiver. AB - A monolithically integrated bacteriorhodopsin-semiconductor phototransceiver is demonstrated for the first time to the authors' knowledge. In this novel biophotonic optical interconnect, the input photoexcitation is detected by bacteriorhodopsin (bR) that has been selectively deposited onto the gate of a GaAs-based field-effect transistor. The photovoltage developed across the bR is converted by the transistor into an amplified photocurrent, which drives an integrated light-emitting diode with a Ga0.37Al0.63As active region. Advantage is taken of the high-input impedance of the field-effect transistor, which matches the high internal resistance of bR. The input and output wavelengths are 594 and 655 nm, respectively. The transient response of the optoelectronic circuit to modulated input light has also been studied. PMID- 15524376 TI - Computational microscopy in embryo imaging. AB - The growth of computing power has greatly improved our ability to extract quantitative information about complicated three-dimensional structures from microscope images. New hardware techniques are also being developed to provide suitable images for these tasks. However, a need exists for synthetic data to test these new developments. The work reported here was motivated by studies of embryo health, but similar needs exist across the field of microscopy. We report a rigorous computer model, based on Maxwell's equations, that can produce the required synthetic images for bright-field, differential interference contrast, interferometric imaging, and polarimetric imaging. After a description of the algorithm, sample results are presented, followed by a discussion of future plans and applications. PMID- 15524377 TI - Real-time three-dimensional optical micromanipulation of multiple particles and living cells. AB - Counterpropagating light fields provide a stationary optical potential well for a Brownian particle. Introducing variability in the relative strengths of the counterpropagating beams allows us to create a more general configuration-the optical elevator. An optical elevator dynamically controls the axial location of the potential minimum where the particle finds a stable equilibrium position. We describe the implementation of multiple real-time reconfigurable optical elevators with the generalized phase contrast method for dynamic manipulation of polystyrene spheres and yeast cells S. cerevisiae in three dimensions. PMID- 15524378 TI - High dynamic range contrast measurements by use of an optical parametric amplifier correlator. AB - A new design of pulse-contrast-measuring correlator, based on optical parametric amplification, is presented. It amplifies the wings of the pulse and allows high contrast measurement with a low-sensitivity detector. The test system demonstrates an ability to measure a dynamic range of >10(11):1. PMID- 15524379 TI - Linear and nonlinear optical studies in photonic crystal alloys. AB - We report on linear transmittance and reflectance as well as on third-harmonic generation in photonic crystal alloys formed by various compositions of polystyrene and poly (methyl methacrylate) colloidal spheres of the same size. These photonic crystal alloys are structurally ordered but contain refractive index disorder and thus provide a random variation of scattering potential. The stopgap shows a monotonic shift in wavelength as a function of composition that can be fitted by assuming an effective dielectric constant for the colloidal spheres. In each alloy a dramatic enhancement of third-harmonic generation is observed, always on the short-wavelength side of the stopgap. PMID- 15524380 TI - Diode-pumped passively Q-switched picosecond Nd:GDxY1-xVO4 self-stimulated raman laser. AB - An efficiency of 8.2% is demonstrated for a diode-pumped passively Q-switched self-stimulated Raman laser with an a-cut mixed vanadate crystal, Nd:Gd0.8Y0.2VO4. At 2.2 W of incident pump power, the self-stimulated Raman laser produces pulses as short as 660 ps at a Stokes wavelength of 1175 nm with 2.7 microJ of energy per pulse at a 66-kHz repetition rate. PMID- 15524381 TI - Starch-based second-harmonic-generated collinear frequency-resolved optical gating pulse characterization at the focal plane of a high-numerical-aperture lens. AB - We report the use of starch as an ideal nonlinear medium with which to perform collinear frequency-resolved optical gating measurements of ultrashort pulses at the focal plane of a high-numerical-aperture (NA) lens. We achieved these measurements by simply sandwiching starch granules (suspended in water) between two coverslips and placing them within the focal plane of a high-NA lens. The natural nonlinear characteristics of starch allow the correct phase matching of pulses at the focal plane of a high-NA lens at different wavelengths. This elegant arrangement overcomes all the complexity and problems that were previously associated with pulse characterization within a multiphoton microscope. PMID- 15524382 TI - Incoherent vector vortex-mode solitons in self-focusing nonlinear media. AB - We suggest a novel type of composite spatial optical soliton created by a coherent vortex beam guiding a partially incoherent light beam in a self-focusing nonlinear medium. We show that the incoherence of the guided mode may enhance, rather than suppress, the vortex azimuthal instability, and we also demonstrate strong destabilization of dipole-mode solitons by partially incoherent light. PMID- 15524383 TI - Method for sensitivity analysis of photonic crystal devices. AB - We present a new method for sensitivity analysis of photonic crystal devices. The algorithm is based on a finite-difference frequency-domain model and uses the adjoint variable method and perturbation theory techniques. We show that our method is highly efficient and accurate and can be applied to calculation of the sensitivity of transmission parameters of resonant nanophotonic devices. PMID- 15524384 TI - Slow light in semiconductor quantum wells. AB - We demonstrate slow light via population oscillation in semiconductor quantum well structures for the first time. A group velocity as low as 9600 m/s is inferred from the experimentally measured dispersive characteristics. The transparency window exhibits a bandwidth as large as 2 GHz. PMID- 15524385 TI - Efficient multiwave mixing in the ultraslow propagation regime and the role of multiphoton quantum destructive interference. AB - We analyze a lifetime-broadened four-state four-wave-mixing (FWM) scheme in the ultraslow propagation regime and show that the generated FWM field can acquire the same group velocity and pulse shape as those of an ultraslow pump field. We show that a new type of induced transparency resulted from multiphoton destructive interference that significantly reduced the pump field loss. Such induced transparency based on multphoton destructive interference may have important applications in other nonlinear optical processes. PMID- 15524386 TI - Expanding amplitude-squeezing bandwidth with off-peak dispersive optical feedback. AB - Weak dispersive optical feedback has been proved both theoretically and experimentally to be able to reduce the intensity noise of semiconductor lasers and allow the generation of amplitude-squeezed states. I propose a novel scheme exploiting the off-peak optical feedback condition that expands the squeezing bandwidth and significantly enhances the repeatability over the most commonly utilized on-peak optical feedback scheme. Using a realistic model, I show that under the zero-frequency-pulling condition the off-peak scheme greatly enhances the squeezing bandwidth. PMID- 15524387 TI - Experimental evidence of 25-fs laser pulse distortion in singlet beam expanders. AB - We report the measurement of spatiotemporal distortions of an ultrashort pulse in singlet beam expanders. With a simple second-order autocorrelator the temporal broadening of the pulse from 23 to 40 fs, due to propagation time difference (PTD), is determined. The delay due to PTD between different parts of the beam is also measured. This effect was theoretically studied for the first time by Bor [J. Mod. Opt. 35, 1907 (1988)]. These experimental results are in good agreement with the calculations of a dedicated three-dimensional ray-tracing program developed to simulate the spatial and temporal transformation of femtosecond pulses in optical systems. PMID- 15524388 TI - Control of atomic ionization by two-color few-cycle pulses. AB - We have studied the ionization of Rydberg atoms by few-cycle radio-frequency pulses and used two-color fields to control the ionization dynamics. We show that the number of times that electrons are emitted during a pulse can be limited and that the duration of the electron emission can be shortened. These results, once they are transposed to the optical domain, may inspire new strategies for the production of single attosecond pulses. PMID- 15524389 TI - Spot-size reduction in terahertz apertureless near-field imaging. AB - We show measurements and calculations of the terahertz (THz) near field of a metal tip with a specially formed, semicircular apex that allows us to identify the separate contributions of the tip apex and shaft to the measured signal. We find that when the tip-crystal distance is not modulated the measured near-field signal is overwhelmed by contributions from the tip shaft, resulting in a relatively large THz spot size. When the tip-crystal distance is modulated, with subsequent lock-in detection at the modulation frequency, only the near-field distribution of the semicircular apex is observed, resulting in a much smaller THz spot size and thus improved spatial resolution. PMID- 15524390 TI - Discrete-mode cancellation mechanism for high-Q integrated optical cavities with small modal volume. AB - A mechanism to reduce radiation loss from integrated optical cavities without a complete photonic bandgap is introduced and demonstrated. It is applicable to any device with a patterned substrate (including both low and high index-contrast systems), when it supports discrete guided or leaky modes through which power escaping the cavity can be channeled into radiation. One then achieves the associated increase in Q by designing the cavity such that the near-field pattern becomes orthogonal to these discrete modes, therefore canceling the coupling of power into them and thus reducing the total radiation loss. The method is independent of any delocalization mechanism and can be used to create high-Q cavities with small modal volume. PMID- 15524391 TI - [Molecular analysis of methicillin-resistant staphylococci isolated from hospital patients and in the out-patient clinic]. AB - In this study, a molecular analysis of the methicillin-resistant coagulase negative staphylococci strains was performed. The obtained results of the biochemical and drug resistant pattern investigations were insufficient to assess the relationship between the strains. Therefore genotyping by the restriction fragment length polymorphism analysis (PCR-RFLP) method was performed. Analyzed strains characterized presence of the mecA gene-PCR products. The PCR products were digested with DraI and TasI, and the fragments separated by agarose gel electrophoresis. Typing of the methicillin-resistant gene using PCR-RFLP showed that all MRCNS strains possess an identical restriction pattern of the mecA gene. This identical restriction pattern of the mecA gene in investigated strains may suggest an easy transfer of this gene between different staphylococci species and lead to the spreading of methicillin-resistant among hospital strains. Furthermore performing the comparison of different phenotype and genotype methods has shown that the PCR-RFLP method is quick and reliable, enabling the detection and estimation of the relationship between MRCNS strains. PMID- 15524392 TI - [Drug resistant coagulase-positive staphylococci strains in food]. AB - From the 1572 food samples, examined in Microbiology Department of Frozen Food Industry Research Laboratory in Lodz, 79 (5.0%) coagulase-positive staphylococci strains were isolated. All the strains were sensitive to vancomycin, gentamycin and chloramphenicol. Only individual staphylococci strains were resistant to erythromycin (1.3%), lincomycin (2.5%) and ciprofloxacin (2.5%). 20.3% strains, isolated mainly from raw meat, were resistant to doxycycline and 6.3% to oxacillin. 38.0% of coagulase-positive staphylococci strains had positive results of cefinase test. One strain isolated from minced meat was resistant to methicillin and at the same time it was producing beta-lactamases. PMID- 15524393 TI - [Coagulase-negative staphylococci in municipal waste]. AB - The total of 259 coagulase-negative staphylococci isolates gained from aerosols created during dumping and utilisation of municipal waste were investigated. Species and subspecies identification allowed to determine predominance of novobiocin-resistant species which primarily colonise rodents or other animals. However, about 30% of isolates were species originating from humans. Metabolic properties of the studied group of isolates were examined, as well as their sensitivity to a set of 8 commonly used antibiotics: amoxicillin, erythromycin, tetracycline, clindamycin, chloramphenicol, gentamicin, trimethoprim/sulfamethoxazole and ciprofloxacin. Many isolates resistant to erythromycin (19%), tetracycline (13%) and a considerably smaller group resistant to clindamycin (6.2%), chloramphenicol (2%) and trimethoprim/sulfamethoxazole (1%) were found. It appeared that processes of occurring at the time of dumping and utilisation of waste affect the prevalence of only some features. The examined metabolic features were found to be relatively stable and no changes were observed, which would indicate the tendency of adaptation to existence in the inanimate matter environment. Only isolates with active nitrate reductase were more frequently detected. The increase in frequency of occurrence of isolates resistant to tetracycline and chloramphenicol and simultaneous elimination of isolates resistant to other antibiotics were observed. PMID- 15524394 TI - [Citric acid as a siderophore of enterococci?]. AB - In the pool of 70 enterococcal strains of the genus Enterococcus 61.4% released citrate into the medium. This metabolite has occurred more frequently in E. faecium strains. There was no correlation between hydroxamate siderophores production and citrate releasing. Only nine (10, 3%) of 70 strains have used Fe3+ dicitrate complex as iron sources. Iron restricted condition causing moderate inhibition of growth have not increased citrate releasing. When iron deficiency has caused stronger growth inhibition, E. faecalis strains did not release citrate and E. faecium strains its smaller amounts. The resting cells grown in iron-restricted condition have incorporated 59Fe3+ complexed by citrate more active than cells grown in the medium with excess of iron. So, citrate has not been a siderophore in enterococci. PMID- 15524395 TI - [Application of released proteins (RPs) of Yersinia enterocolitica for serological diagnosis of yersiniosis. II. Gene recombination for production of the YopD protein]. AB - The aim of this study was to evaluate the usefulness of gene recombination technique using the pET-30 Ek/LIC expression vector for production a 36 kDa released protein called YopD and evaluate of this purified protein as antigen in serodiagnosis of yersiniosis. Protein YopD of Y. enterocolitica was expressing in Escherichia coli BL21 (DE3) using the pET-30 Ek/LIC expression vector. Purification of the expressed enzyme from suspensions of E. coli cells treated with Bug Buster Protein/Extraction Reagent was accomplished by immobilised metal (Ni2+) affinity column chromatography (His-trap). The IgM, IgG and IgA class antibodies to YopD were measured in 100 serum samples collected from patients suspected for yersiniosis and 100 blood donors. The obtained results were compared to the results of ELISA with released proteins isolated from the culture of Y. enterocolitica supernatant under calcium deficient conditions and commercial ELISA with recombinant released proteins. A very high (94.0-100.0%) specificity and good sensitivity (55.2-80.4%) were displayed by the ELISA with YopD in relation to other two ELISA. The results of our study showed that recombinant YopD protein purified by chromatography of bio-affinity may be used in serodiagnosis of yersiniosis as a high specific antigen free of Yersinia lipopolysaccharides. PMID- 15524396 TI - [Comparison of the results of disc diffusion methods applied for detection of ESBL-positive strains of gram-negative rods]. AB - Examinations were undertaken to compare the results of disc diffusion tests applied for detection of strains producing extended-spectrum beta-lactamases (ESBLs). A total of 120 clinical strains were used in experiments. These strains were determined as ESBL-positive on the basis of consistent results of two methods: the double disc synergy test (DDST) according to Jarlier et al. (1988) and the diagnostic disc test (DD, version CPD/CD 01) according to Appleton (1999). In the next step examined strains were analysed in two further tests, which are variants of DD method: CAZ/CD 02 test with discs containing ceftazidime and ceftazidime/clavulanic acid, and CTX/CD 03 test with the use of cefotaxime and cefotaxime/clavulanic acid discs. ESBL-positive strains first of all belonged to the species E. coli and K. pneumoniae. In the case of seven analysed strains consistent results of determinations were not obtained with the use of different disc diffusion methods. Application of several disc diffusion methods to determine ESBL-positive strains of gram-negative rods increases the probability of their proper identification. PMID- 15524397 TI - [Hydrophobic properties of gram-negative rods colonizing upper respiratory tract of healthy people]. AB - The cell surface hydrophobicity is one of the non specific factors of adhesion influencing the ability of microorganisms to colonize nasopharynx. The aim of this paper was to evaluate via salt aggregation test (SAT) the cell surface hydrophobicity of 150 strains of gram-negative rods isolated from the throat or/and nasal specimens of healthy people. It has been found that among the nonfermenting rods hydrophobic strains were predominant. In contrast, the isolates of Enterobacteriaceae family were characterized by the distinctive features of the cell surface within particular genera or even species. The obtained results show that, despite differences in cell surface hydrophobicity, numerous species of gram-negative rods have the ability to colonize the mucous membrane of upper respiratory tract. This suggests that the cell surface hydrophobicity is rather a feature of species or genus, but it is not related to the ecological niche of microorganisms in human body. PMID- 15524398 TI - [Hydrogen peroxide produced by Lactobacillus species as a regulatory molecule for vaginal microflora]. AB - A total of 33 strains of Lactobacillus belonging to 9 species, isolated from vagina, were tested for production of hydrogen peroxide. We observed that the following species: L. delbrueckii, L. acidophilus, L. crispatus, L. johnsonii and L. gasseri dominated over other species in secretion of hydrogen peroxide to the growth medium. Concentration of this substance amounted from 0.05 to 1.06 mM (in case of strong aeration the concentration increased up to 1.8 mM). Moreover, killing properties of the pure hydrogen peroxide exerted toward Escherichia coli and Candida albicans were less prominent than these of the supernatants of cultures of Lactobacillus strains producing H2O2. PMID- 15524399 TI - [Characteristics of the Propionibacterium strains isolated from acne patients]. AB - Propionibacterium acnes is a component of physiological flora of human skin. It colonizes the outlets of sebaceous glands and participates in the pathogenesis of inflammatory acne. Acne vulgaris is a common skin disease. It is found in more or less exacerbated form in approximately 85% of adolescent population. The main purpose of the research was to confirm the hypothesis of Propionibacterium bacteria participation in the aetiopathogenesis of acne vulgaris. The researches have proved the presence of Propionibacterium acnes on the surface of the skin both of people with acne-related changes and these with whom such changes were not found. Statistically significant differences were found in the number of P. acnes bacteria per 1 square centimeter of healthy and disease-affected skin as well as in the diversity of biochemical types. The highest number of P. acnes bacteria have been found in fresh changes with visible symptoms of inflammation. In order to confirm the hypothesis of the participation of Propionibacterium bacteria in the aetiopathogenesis of acne, a detailed phenotypical analysis of isolated P. acnes strains have been conducted. Type, biotype, resistance pattern, proteolytic and lipolytic properties have been determined. PMID- 15524400 TI - [Diagnostic value of Gram stain for assessment of vaginal smears during pregnancy]. AB - The main aim of this study was to compare the diagnostic value of Giemsa stained method with Gram stained method for the evaluation of vaginal smears among pregnant women. A study population comprised 111 pregnant between 6 and 30 weeks of gestation. The vaginal smears from every subject was diagnosed according to Giemsa and Gram stained method and micro-organisms were isolated by culture. In 29.3% cases diagnosed as normal flora (2a) on the basis of Giemsa method bacterial vaginosis was detected in Gram stains according to Spiegel's criteria and pathological microflora in concentration > or = 10(5) CFU/ml was cultured among 75.9% of them. Among 31.7% women who had grade 3a (abnormal) in Giemsa stains method normal flora was diagnosed on the basis on Gram's method and from 17.1% pregnant women from this group we did not isolated any pathogens. For evaluation of vaginal smears during pregnancy the Giemsa method should be replaced by Gram stained method. PMID- 15524401 TI - [Comparison of polymerase chain reaction assay and Mycoplasma IST 2 test with culture for detection of infections caused by Ureaplasma urealyticum and Mycoplasma hominis]. AB - The polymerase chain reaction (PCR) technique and commercial Mycoplasma IST 2 test were compared with culture for the detection of U. urealyticum and M. hominis in 173 clinical samples obtained from patients without clinical symptoms from genito-urinary tract. The presence of U. urealyticum was diagnosed by culture in 24 samples, by PCR in 33 samples and by Mycoplasma IST 2 test in 39 samples. The presence of M. hominis was diagnosed in 26 samples only by Mycoplasma IST 2 test--culture and PCR were negative. The study showed the excellent sensitivity (100%) and good specificity (appropriately 94.0% and 90.0%) for U. urealyticum in PCR and Mycoplasma IST 2 test. The discrepancy of results obtained in Mycoplasma IST 2 test and culture as well as in PCR may suggest the over sensitivity of the commercial test for detection of M. hominis. PMID- 15524402 TI - Safrole oxide induces apoptosis in A549 human lung cancer cells. AB - 3,4-(Methylenedioxy)-1-(2',3'-epoxypropyl)-benzene (safrole oxide) was synthesized in the authors' laboratory. To investigate the effects of safrole oxide on the growth and apoptosis of A549 human lung cancer cells, the authors treated the cells with safrole oxide, 112.36 to 449.44 micromol/L, for 24 to 48 hours. The results showed that the drug led A549 cells to apoptosis and blocked cell cycle completely at G1 phase and partly at G(2)-M phase. To further study the correlated mechanism, the authors examined P53 and H-Ras protein expressions by using immunofluorescence assay. They found that the expression of P53 was dramatically up-regulated but the expression of H-Ras was hardly affected by safrole oxide, 224.72 micromol/L, within 24 hours. Taken together, these results revealed that safrole oxide could induce apoptosis of A549 cells and suggested that safrole oxide might perform its function by blocking cells completely at G1 phase and partly at G(2)-M phase, and also by up-regulating the expression of P53 protein. These findings would raise exciting possibilities for cancer therapy in future. PMID- 15524403 TI - Effects of N-acetylcysteine on oxidant-antioxidant balance in oleic acid-induced lung injury. AB - The antioxidant and anti-inflammatory properties of N-acetylcysteine has been documented in many experimental lung injury models. Because intravenous injection of oleic acid induces histopathologic changes similar to those seen in human acute lung injury or acute respiratory distress syndrome, the authors evaluated the effects of N-acetylcysteine (NAC) on oxidative stress and lung damage in an oleic acid (OA)-induced lung injury model. Thirty-five rats were divided into 5 groups as sham, NAC, OA, pre-OA-NAC, and post-OA-NAC. Lung damage was induced by intravenous administration of oleic acid. Pre-OA-NACgroup received intravenous (IV) N-acetylcysteine 15 minutes before oleic acid infusion and post-OA-NAC group received IV N-acetylcysteine 2 hours after oleic acid infusion. In both of the N acetylcysteine treatment groups, blood and tissue samples were collected 4 hours after oleic acid infusion, independent from the time of N-acetylcysteine infusion. In other groups, blood and tissue samples were collected 4 hours after ethanol, NAC, or OA infusions. Serum myeloperoxidase activity, total antioxidant capacity, malondialdehyde levels, and lung tissue Na+ - K+ ATPase activity were measured and light microscopic analyses of lung specimens were performed. The administration of N-acetylcysteine significantly restored Na+ - K+ ATPase activity and total antioxidant capacity levels and ameliorated lung architecture. N-acetylcysteine has been shown to have some attenuating effects in experimental animal studies. However, further investigations are necessary to suggest N acetylcysteine as a treatment agent in critically ill patients with lung injury. PMID- 15524404 TI - p21(Waf1/Cip1) regulates proliferation and apoptosis in airway epithelial cells and alternative forms have altered binding activities. AB - p21(Waf1/Cip1) plays central roles in proliferation, differentiation, and apoptosis. Alterations in the expression and subcellular localisation of p21 occur during several lung diseases but the roles of p21 in the lung epithelium are unknown. The effects of p21 on proliferation and apoptosis in mouse airway epithelial cells (AECs) were examined using p21-null mice. AECs isolated from p21 null mice had increased proliferation and apoptotic rates compared to AECs from wild-type mice. Alterations in the subcellular localization of the cell cycle regulatory proteins p27, PCNA, and p53 were also evident in p21(-/-) cells. The nuclear and cytoplasmic forms of p21 present in AECs were also examined. Full length p21 (20 kDa) was detected in nuclear fractions but a C-terminal truncated form (17 kDa) of p21 was present in cytoplasmic fractions. The binding activities of truncated p21 were altered compared to full-length p21. Although the latter was complexed with PCNA, Cdk2, Cdk4, Cdk6, cyclin D3, and cyclin E, truncated p21 was bound only to Cdk4 and cyclin D3. In conclusion, p21 regulates proliferation and protects against apoptosis in AECs. In addition, different forms of p21 are present in AECs and the subcellular localization of these forms reflects differences in p21 activity. PMID- 15524405 TI - Neuregulin-1 induces branching morphogenesis in the developing lung through a P13K signal pathway. AB - Neuregulin-1 (NRG-1) induces signal transduction through the activation of its receptor, a heterodimer of human epidermal growth factor receptors 2 and 3 (HER2/HER3). Signal transduction through this receptor/ligand system plays a critical role in the developing heart, mammary gland, and nervous systems. Previous studies showed that NRG-1-induced HER2 activation resulted in pulmonary epithelial cell proliferation in the human fetal lung. The authors hypothesized that NRG-1 further contributes to lung development and maturation by inducing branching morphogenesis. In the present study, the authors show that NRG-1, HER2, and HER3, but not HER4, are expressed in the developing mouse lung. Addition of NRG-1 to fetal lung explants increased lung branching morphogenesis by 32% (P < .05). This increase in branching was blocked by 2C4, an antibody directed against HER2 that inhibits its dimerization and subsequent NRG-1-induced signal transduction. To gain an understanding of the intracellular signaling pathways involved in NRG-1-induced branching morphogenesis, the authors specifically blocked the phosphatidylinositol-3 kinase (PI3K) and mitogen activation protein kinase (MAPK) pathways. Inhibition of PI3K signaling significantly decreased NRG 1-induced branching morphogenesis (P < .05). Inhibition of NRG-1-induced MAPK activation had no effect on explant branching morphogenesis. These data suggest that NRG-1, binding to the HER2/HER3 heterodimer receptor complex, induces pulmonary branching morphogenesis through HER2 activation of the PI3K pathway. PMID- 15524406 TI - Direct visual instillation as a method for efficient delivery of fluid into the distal airspaces of anesthetized mice. AB - Although several methods have been used to deliver fluid into the distal airspaces of the lung, the efficiency of these methods has been variable. Therefore, the authors have modified prior techniques to design a better method for direct visual instillation (DVI) of fluid into the trachea and compared its efficiency with two commonly used methods: nasal inhalation and invasive intratracheal instillation (delivery of the instillate by needle puncture of the trachea). The results showed that this method (DVI) can deliver fluid efficiently into either both lungs or into a single lung. Using an 131I-albumin labeling technique, DVI resulted in 92 +/- 1% retention of the labeled albumin in the lungs 1 hour after instillation, significantly greater than nasal inhalation (48 +/- 3%, P < .01) and invasive intratracheal instillation (77 +/- 3%, P < .05). Also, when bacteria (Escherichia coli) were instilled with the DVI method, the severity of gram-negative pneumonia was greater (6.5 +/- 0.5 g water/g dry weight) compared to delivery by nasal inhalation (5.5 +/- 0.4 g water/g dry weight, P < .05) or by invasive intratracheal instillation (5.9 +/- 0.4g water/g dry weight, P < .05). The authors conclude that DVI is more efficient than nasal inhalation and invasive intratracheal instillation for delivering experimental fluids into the distal airspaces of anesthetized mice. This method should be valuable for experimental lung studies in mice. PMID- 15524407 TI - [Effect of pregnancy on diabetic vascular complications]. AB - OBJECTIVES: There are still controversies referring to pregnancy influence on the development of vascular complications in type 1 diabetes. MATERIALS AND METHODS: 227 diabetic women were taken into the study (109 who gave at least one birth (GB) and 118, who never were pregnant (NB)). All of patients started to be diabetic before the 15 years of age. We compared the development of vascular complications (VC) (hypertension-RR, proteinuria (P), retinopathy (R) and the age of patients, HbA1C, serum creatinine (Cr), creatinine clearance (CrCl) between the GB and NB groups. We also compared the development of VC in relation to risk factors (RF): HbA1c and duration of diabetes. RESULTS: Women who gave birth were significantly older and developed diabetes earlier than childless women. We didn't find any significant difference in duration of diabetes, HbA1c, mean blood pressure (BP), Cr and CrCl between these two groups. In the GB group in comparison to NB we noticed higher frequency of RR (11.1% vs 6.8%, OR 1.7), proliferative retinopathy (9% vs 6.8%, OR 1.4) and laser therapy (5.5% vs 3.4%, OR 1.6). In the next step we analyzed the development of VC in relation to risk factors: duration of diabetes (less or more than 20 years) and HbA1c levels (below and above 8.0%). Patients, who developed diabetes before 20 years of age were significantly older (36.3 vs 31.2 years p < 0.001), duration of diabetes in this group was longer (26 vs 14 years p < 0.001), Cr was higher (0.9 mg/dl vs 0.8 mg/dl p < 0.05)and CrCl was lower (73 mg/ml vs 86 mg/ml p < 0.05). In this group the frequency of proteinuria as well as retinopathy were higher (18.2% vs 7.9%, OR 2.59) (52% vs 9.2%, OR 10.6). We didn't find the difference in HbA1c concentration between the groups with different duration of diabetes. Higher HbA1c presented no relation to the age of patients, but correlated with lower frequency of patients without retinopathy (60.3% vs 66% OR 0.68). In this group laser therapy had to be performed more frequent (6.8% vs 1.8% OR 6.1). CONCLUSIONS: Pregnancy in diabetic patients with good metabolic control doesn't seem to be a risk factor for developing vascular complications. These complications are still strongly correlated with duration of diabetes. PMID- 15524408 TI - [IL-1beta and PGE2 production in whole blood and gingival fluid in women with periodontitis and preterm low birth weight]. AB - DESIGN: The aim of the investigation was to evaluate of IL-1beta and PGE2 concentrations in gingival fluid, whole blood as well as IL-1beta, PGE2 production after Escherichia coli lipopolysaccharide stimulation in whole blood in women with preterm low birth weight (PLBW), as compared to the control group. MATERIAL AND METHODS: A case-control study of 88 postpartum women aged 17 to 39 was performed. The case group consisted of 52 women with PLBW and the control group consisted of 36 women giving birth in time. Concentration of inflammatory mediators in gingival fluid, blood serum and IL-1beta, PGE2 production in whole blood after bacterial lipopolysaccharide stimulation were determined by means of immunoenzymatic method. RESULTS AND CONCLUSIONS: The levels of IL-1beta and PGE2 in gingival fluid were significantly higher in all PLBW mothers (also PLBW primiparous) than in the control group. In addition in the primiparous with PLBW group significantly higher PGE2 concentration in blood serum was found compared to the primiparous controls. There were no significant differences between women with PLBW and the controls together with a significantly higher production of IL 1beta and PGE2 in whole blood after LPS stimulation in women with periodontitis and gingivitis compared to subjects with healthy periodontium. Such findings suggest that inflammatory mediator synthesis is mainly result of specific cells exposition to bacterial products. Therefore it seems that more frequent occurrence of the phenotype of hyperactive cells that synthesise these mediators is not responsible for PLBW. PMID- 15524409 TI - [Clinical analysis of pregnancy complicated by hypertension in the material of the Department of Obstetrics and Perinatology of the Pomeranian Academy of Medicine]. AB - OBJECTIVES: The aim of this study was retrospective analysis of arterial hypertension during pregnancy in the Department of Obstetrics and Perinatology of Pomeranian Academy of Medicine in Szczecin, and severe PIH intensive obstetrical care optimization. DESIGN: The retrospective analysis of 81 pregnancies complicated by arterial hypertension in the years 1995-2000 was performed. MATERIAL AND METHODS: The retrospective analysis of 81 pregnancies was performed. Patients were divided into two groups. In the first pregnancy was ended by caesarean section (n = 43), in the second by vaginal delivery. The mean gestational age, the way of delivery, accompanying diseases, uric acid levels, short term variability, Dawes - Reedman's criteria, presence of decelerations in CTG, Doppler PI, S/D, RI parameters in umbilical artery and cerebral arteries, presence of AEDVF and REDVF in umbilical artery were analyzed. RESULTS: Caesarean section was performed in 53.1% of all cases, in 46.9% vaginal delivery took place. Absolute range of short-term variability was more often less than 6 ms in caesarean section group (41.9%). Uric acid level was also higher in caesarean section group (p = 0.000194) CONCLUSIONS: 1. The caesarean rate in pregnancies complicated by arterial hypertension is over 50% and during severe PIH is approximately 100%. 2. Caesarean section takes place before estimated delivery date and indications to it are in most cases connected with a main disease 3. Short-term variability is lower among patients from caesarean section group. 4. Uric acid level is a relevant parameter of a degree of arterial hypertension, and the level is higher in first group. 5. Doppler velocimetry of umbilical artery and middle cerebral artery are valid part of obstetrical care among patients with PIH. PMID- 15524410 TI - [Vitamin D receptor polymorphism in the group of postmenopausal women with low bone mineral density]. AB - OBJECTIVES: Osteoporosis is a multifactoral disease with aetiology depending from the hormonal, environmental, and genetic factors. One of the suggested candidate gene involved in the pathogenesis is the polymorphic gene encoding for vitamin D receptor (VDR). VDR polymorphism was connected with bone mineral density (BMD) and correlated with onset of osteoporosis. The goal of our study was to determine the role of BsmI polymorphism of VDR gene in the group of postmenopausal women with low bone mineral density. MATERIALS AND METHODS: We have analysed the group of 34 postmenopausal women. The DNA analysis was performed using PCR/RFLP (polymerase chain reaction/restriction fragment length polymorphism) assays. RESULTS: In our investigation we have observed statistically higher frequency of B allele (48.5% vs. 41.2%), the lower frequency of b allele (51.5% vs. 58.8%) and bb genotype (8.8% vs. 42.5%) in the investigated group of postmenopausal women with low BMD. CONCLUSIONS: Our observations could suggest the important role of B allele of the VDR gene in the pathogenesis of osteoporosis in the group of women with low mineral density and possible protective role of b allele in this disease. PMID- 15524411 TI - [Evaluation of the quality of life of women in the climacteric period]. AB - OBJECTIVES AND STUDY DESIGN: Evaluation of quality of life of women in the climacteric period with the use of an individually developed questionnaire of a transitory period. Evaluation of the effect of Hormonal Replacement Therapy (HRT) on health related quality of life. MATERIALS AND METHODS: A group of 190 women in the perimenopause period was subject to an analysis. The criterion of qualification for the investigated group was medical history of a patient, which revealed climacteric symptoms and age of women (from 45 to 60 years old). The patients were divided into two main groups depending on the use of HRT The first group--investigated group - adhibiting HRT consists of 76 patients, the second one--control group--114 patients. Within each of the groups, 5 subgroups were isolated basing on the moment of the menopause occurrence. The research tool was an individually developed questionnaire consisting of SF-36, Green Scale and Beck Depression Scale. RESULTS: Between the group subject to HRT and the control group, a statistically significant difference (p < 0.05) was observed concerning the quality of life parameters, intensification of depression connected with climacteric and symptoms of the climacteric syndrome. CONCLUSIONS: 1. The application of HRT in women after menopause contributes to an improvement of the general quality of life, of mood and vitality. 2. Women subject to the therapy constitute a minority. Thus, HRT should be promoted in order to improve the life quality of women in that period so difficult for them. 3. The transitory period questionnaire is a good work tool, which allows to determine the necessity of the therapy application and to monitor its course. PMID- 15524412 TI - [Staphylococcal toxic shock syndrome]. AB - [A case of menstrual toxic shock syndrome (TSS) in a woman aged 21 is reported. Diagnosis was made on clinical basis and was followed by successful treatment] PMID- 15524414 TI - [Periodontitis and preterm low birth weight]. AB - During the past decade an issue of influence of periodontitis on developing of systemic social diseases with polyfactorial etiology (atherosclerosis, myocardial infarction and stroke, diabetes, lung diseases) was taken up. Several researches have also proved that periodontitis could be an independent risk factor for preterm low birth weight (PLBW). The nature of this connection is not clear yet. The aim of this article is to show the current evidence from experimental studies in animals and human case-control studies on the relationship between these pathologies. Potential etiopathogenic mechanisms for periodontitis and preterm low-birth weight mothers have also been demonstrated. It's possible that treatment of periodontitis may reduce the risk of preterm birth. PMID- 15524413 TI - [Vascular complications and their effect on fetal-maternal outcome and therapeutic approach in diabetic women]. AB - Metabolic disturbances related to glucose intolerance and hyperglycemia are known to affect the health and life of women planning the pregnancy. They also are capable of impairing the development of fetuses and newborns. Remarkable advances have been made in recent years in clarifying the metabolic processes that occur during pregnancy and their effect on intrauterine fetal development. Clinicians have become aware of the observations, that maternal diabetes and the occurrence of complications are related to the duration of the disease, to the grade of hyperglycemia and to the time of pregnancy when hyperglycemia occurred. Since the last decade, numerous epidemiological studies have confirmed that a good balanced diabetes mellitus in pregnant women leads to the fruitful fetal development and contributes to reducing maternal mortality. The aim of this study was to discuss the update guidelines for managing diabetic patients with vascular complications before and during the course of pregnancy. PMID- 15524415 TI - [Genetic risk factors of osteoporosis--vitamin D receptor gene polymorphism]. AB - The aetiology of osteoporosis is not fully understood but it was suggested that the genetic factors could play an important role. The many studies have presented the key role of the polymorphism of the gene encoding for vitamin D receptor (VDR). The modern concepts about VDR and osteoporosis were summarised in this review. PMID- 15524416 TI - [The stress level and structure in parents awaiting babies from high risk pregnancies]. AB - OBJECTIVES: The analysis of stress in patients with high risk pregnancies. DESIGN: The purpose of this research was analysis of stress level and structure in patients awaiting the baby from high risk pregnancies. MATERIALS AND METHODS: 50 married couples were recruited to the study: 14 with threatened abortion, 18 with impending preterm delivery and 18 with a chronic disease (preeclampsia or diabetes). Trauma Symptoms Inventory (J. Briere) was implemented in this research. RESULTS: Women exhibited higher levels of anxious arousal (AA). They revealed more frequently lowered mood and depression (D), as well as anger and irritability (AI). The clinical type of the threat seems to be the factor modifying the level of experienced stress. The highest level of stress occurred in those couples in which the women suffered from a chronic disease (preeclampsia, diabetes). CONCLUSIONS: Threatened pregnancy is a trigger for psychological stress in parents. Both women and men reveal increased behaviours directed to reduction of emotional tension. The level of experienced stress is most heavily affected by the type of threat. The symptoms of stress are similar in both parents in a group with a particular diagnosis. PMID- 15524417 TI - [Lymphangioma--prenatal and perinatal aspects--Part one]. AB - Lymphangioma or cystic hygroma is a congenital malformation of the lymphatic system which has been commonly associated with fetal aneuploidy, hydrops, structural malformations and intrauterine death. In this paper we would like to report two cases of lymphangioma diagnosed prenatally in the third trimester in the fetuses with normal karyotype, normal NT in the first trimester and without other structural anomalies and with good perinatal outcome. PMID- 15524418 TI - [Lymphangioma--the care and treatment in neonates and babies--Part two]. AB - Lymphangioma or cystic hygroma (CH) in neonates is a rare congenital malformation (1:12000 live births), its etiology is unknown. It often occurs in head and neck region. Initial diagnosis based on physical examination is next to be confirmed by MR and CT imaging. The authors presented with diagnosis and treatment of two babies with congenital cystic hygromas of face, neck and thorax. Although in cystic hygroma the surgery still remains as the treatment of choice, the OK-432 sclerotherapy seems to be very promising in huge CH lesions. PMID- 15524419 TI - [Procalcitonin and C-reactive protein as a markers of neonatal sepsis]. AB - OBJECTIVES: Sensitive, reliable and early parameters of bacterial infection are extremely valuable in diagnosis of nosocomial infections in neonatal intensive care unit. In this study procalcitonin (PCT) and C-reactive protein (CRP) were evaluated for their diagnostic relevance in neonatal late onset sepsis. DESIGN: Clinical study MATERIALS AND METHODS: We analysed inflammatory parameters in 48 newborn infants admitted to the Intensive Care Unit of Institute of Paediatrics in Lodz who suffered from nosocomial sepsis. They were sampled for PCT and CRP levels at the time of the onset of signs and 24 hours later. CRP was determined by an nephelometric method and PCT was determined by an immunoluminometric assay. RESULTS: At the onset of Gram negative sepsis 14 from 17 contaminated newborns had significantly increased CRP levels and 15 of them had increased levels of PCT After 24 hours 100% of them had elevated PCT and CRP levels. At the onset of Gram positive sepsis only 18 from 31 neonates with positive blood culture had increased CRP levels and 28 of them had elevated concentrations of PCT. This difference was statistically significant. After 24 hours 26 of them had elevated CRP and 100% had increased PCT concentrations--this difference was not significant. CONCLUSION: Measurement of procalcitonin concentrations may be useful for early diagnosis of late onset sepsis in neonates. PMID- 15524420 TI - [Echocardiographic abnormalities in infants with heart murmur]. AB - OBJECTIVES: Congenital heart diseases are very serious problem of current perinatology. Early detection of congenital heart diseases and new possibilities of their treatment have decreased mortality rate in neonates. THE AIM OF THE STUDY: Was to present the reasons of the heart murmurs found in newborns, based on postnatal echocardiography. MATERIAL AND METHODS: The study group consisted of 107 neonates born in The Obstetrics and Gynaecology Clinic of Medical University in Warsaw in the period from 1st October, 2001 until 30th September, 2002, found to have murmurs during the observation in The Neonatology Department. RESULTS: Heart murmurs were detected in 107 infants out of 1291 born in The Clinic in this period. Out of the study group, echocardiography results were normal in fourteen children, and in ninety-three patients the echocardiography revealed one of the following abnormalities: persistent foramen ovale (FO), persistent ductus arteriosus (PDA), ventricular septum defect (VSD), insufficiency of tricuspid valve (IT) and others. CONCLUSIONS: The most frequently observed abnormality in echocardiography in infants with heart murmur was persistent foramen ovale. The rate of heart abnormalities was the highest in infants with birth weight below 1500 G. PMID- 15524421 TI - [Occurrence of fungal pathogens in the delivery rooms of a hospital obstetrics department]. AB - OBJECTIVES: Assessment of the incidence of fungal pathogens in air of the operating rooms from one of the hospitals in Bialystok. MATERIAL AND METHODS: Investigations were conducted in selected rooms of obstetrics department. Material for mycological studies was air sampled at the entrance of hospital building, the entrance to operating room, hall and selected rooms of the department. Fungi were identified using the standard microbial procedures: The monitoring of airborne fungi pollution was done using SAS SUPER 100 (pbi international). Classification of the isolated fungi was done with an accordance to the current procedures. RESULTS: In the air of 16 rooms of obstetrics department different numbers of fungal colonies from 0 to 560 CFU/1000L of air were isolated. Fungi were not isolated from the air samples of preparing, septic, operating and family deliveries rooms. The highest number of fungi colonies were isolated at the entrance the hospital. The following fungal pathogens isolated from the air were: Candida albicans. non-Candida albicans, Penicillium species, Cladosporium species and Aspergillus species. CONCLUSIONS: 1. The different number of fungal colonies was found depending on type of the hospital room. 2. The highest number of fungal colonies was isolated from the air samples of patients rooms 3. No fungal colonies were isolated from the septic, operating and family deliveries rooms 4. The main fungal pathogen isolated from the air samples was Candida albicans. PMID- 15524422 TI - [HPV, Chlamydia trachomatis and genital mycoplasmas infections in women with low grade squamous intraepithelial lesions (LSIL)]. AB - OBJECTIVES: The purpose of the study was to determine the correlation between Chlamydia trachomatis, urogenital mycoplasmas and Low Grade Squamous Intraepithelial Lesions (LSIL) in women with and without Human Papilloma Virus (HPV) infection. MATERIALS AND METHODS: The specimens were tested for: carcinogenic HPV by the Hybrid Capture I Assay, Chlamydia trachomatis antigen by direct immunofluorescence, urogenital mycoplasmas by Mycoplasma IS test. Cytological smears were classified according to the Bethesda system. RESULTS: High-oncogenic HPV types, Chlamydia trachomatis and mixed infections with Mycoplasma hominis and Ureaplasma urealyticum in patients with LSIL occur significantly more frequently comparing to women without dysplasia. Statistically significant correlation between C. trachomatis and presence of HPV was determined. In HPV negative women there was no correlation between C. trachomatis and LSIL. CONCLUSION: In women infected with HPV, especially high-oncogenic types, C. trachomatis test should be included in diagnostic-therapeutic routine scheme. PMID- 15524423 TI - [The influence of HRT on exercise performance in women with coronary artery disease]. AB - OBJECTIVES: The aim of this study was to evaluate the effect of hormone replacement therapy (HRT) on exercise tolerance and ischemia during treadmill exercise test. DESIGN: Prospective, cohort study. MATERIALS AND METHODS: Fifty six postmenopausal women with proven coronary artery disease (CAD) were enrolled into the study (31 were assigned to HRT and 25 were matched as a control group). Clinically both groups were comparable. All patients underwent treadmill exercise test at the beginning and after 12 months of follow up. RESULTS: After 12-months of follow-up exercise tolerance increased significantly and similarly in both groups: duration time (sec): HRT 347 +/- 142 to 401 +/- 113; control 328 +/- 112 to 368 +/- 120; workload (MET): 6.6 +/- 2.1 to 6.8 +/- 2.0 and 6.1 +/- 1.9 to 6.8 +/- 1.7; time to onset of ischemia (sec): 256 +/- 116 to 293 +/- 120 and 238 +/- 87 to 299 +/- 133. Number of leads with ST changes and total sum (mm) of ST depression in all leads did not change significantly. All above parameters were comparable at baseline and after 12 months follow-up between both groups. CONCLUSIONS: Short term, combined, transdermal HRT does not improve exercise tolerance nor coronary ischemia during exercise test in women with CAD. PMID- 15524424 TI - [Sibutramine administration in polycystic ovary syndrome treatment]. AB - Obesity is considered important factor contributing development of PCOS. Insulin resistance, low level of serum SHBG and activation of pituitary-adrenal axis plays a major role in pathogenesis and symptoms of PCOS. The achievement of weight loss is an important factor in treatment of PCOS, but difficulties with adherences to the dietary regiment discourage physicians to use this method. In our paper we would like to show the effectiveness of such treatment and pay attention to possibilities of drug facilities in dietary approach in treatment of PCOS coexisting with obesity. PMID- 15524425 TI - [Endometrial brush biopsy in monitoring endometrial cytology during hormone replacement therapy]. AB - During the past few decades, a variety of diagnostic methods for endometrium direct sampling have been developed. One of these new techniques is endometrial brush biopsy (uterobrush). The principal aim of uterobrush is early detection of endometrial carcinomas and their precursors, but it may be also used to follow up the patients undergoing hormone replacement therapy (HRT). So far, there are only few clinical reports evaluating uterobrush as a technique useful in monitoring endometrial cytology during HRT. In this study the world literature on using brush biopsy to monitor endometrial condition of women receiving HRT has been reviewed. PMID- 15524426 TI - [Immunophysiology of normal pregnancy. Part I. Immunoregulatory mechanisms inside the maternal-fetal interface]. AB - Contemporary opinions concerning the function of decidual NK cells, T and B lymphocytes as well as macrophages and neutrophils in normal pregnancy were presented in the paper. Maintenance of proper Th1/Th2 balance concerning cytokines produced by trophoblast, decidua and decidual infiltrating immunological cells was also precisely described. PMID- 15524428 TI - Concise total synthesis of (-)-muricatacin by tandem ring-closing/cross metathesis. AB - A strategy for the synthesis of chiral 5-(1-hydroxyalk-2-enyl)-5H-furan-2-ones and its application to the total synthesis of (-)-muricatacin, in four steps and 37% overall yield from (R,R)-hexa-1,5-diene-3,4-diol, are described. The key synthetic step in this approach is a highly regioselective and stereoselective tandem ring-closing/cross metathesis reaction in which both lactone formation and alkyl chain extension are accomplished in an efficient one-pot process. PMID- 15524427 TI - Novel sesterterpenes from Leucosceptrum canum of nepalese origin. AB - Two novel sesterterpenoids, leucosesterterpenone (1) and leucosesterlactone (2), with novel skeleta were isolated from the hexane extract of the medicinal plant, Leucosceptrum canum. Their structures were established by the analysis of NMR data and the single-crystal X-ray diffraction of compound 1. Compounds 1 and 2 were found to exhibit activity against prolylendopeptidase (PEP). PMID- 15524429 TI - Highly enantioselective phenylacetylene additions to ketones catalyzed by (S) BINOL-Ti complex. AB - The readily available and inexpensive (S)-BINOL ligand in combination with Ti(O(i)Pr)(4) is an effective chiral catalyst for the catalytic asymmetric addition of alkynylzinc to unactivated simple ketones. Good to excellent enantioselectivities were achieved. No previous case has been reported successfully using BINOL to catalyze the addition of phenylacetylene to unactivated ketones, and thus the utility of BINOL in asymmetric catalysis is expanded. PMID- 15524430 TI - Pyridine-capped, oligomeric (o-phenyleneethynylene)s. AB - The synthesis, optical properties, and solid-state structures of a series of o (aryleneethynylene) oligomers containing three to seven aryl units is described. The main chain in all examples is composed of between one and five 4,5 dimethoxybenzene rings which are capped on each end by 2-pyridyl rings. These oligomers demonstrate pH-dependent emission and absorption spectra. Single crystals were obtained from the aryleneethynylenes containing three to five arene rings. These single crystals were investigated by X-ray diffraction. PMID- 15524431 TI - Oxazatricyclic noradamantanes: stereocontrolled synthesis of functionalized scopolines, related cage molecules, and drug leads. AB - Scopolines 4 and the noradamantane scaffold are accessible from 8 oxabicyclo[3.2.1]oct-6-en-3-ones such as 6 by a concise route involving introduction of an axial amino nitrogen at C3, epoxidation, and cyclization. The resulting cage molecules are versatile drug leads. PMID- 15524432 TI - Synthesis of the azaspiracid-1 trioxadispiroketal. AB - The de novo analysis, design, and synthesis of the azaspiracid-1 trioxadispiroketal system is described. A revised structural model was developed on the basis of an independent analysis of the NMR spectral data of the natural product that fit all of the data and the thermodynamically favored spiroketal paradigm. This model was then tested via synthesis using a novel trioxadispiroketalization process and supported by spectroscopic correlation. PMID- 15524433 TI - Synthesis and lectin binding ability of glycosamino acid-calixarenes exposing GlcNAc clusters. AB - Novel calix[4 or 8]arene-based glycoconjugates exposing terminal N-acetyl-D glucosamine clusters have been synthesized using amino acid-calixarenes as building blocks. The obtained glycosamino acid-calixarenes 9b-14b have lectin binding ability and amplified inhibitory effects on erythrocyte agglutination induced by wheat germ (Triticum vulgaris) agglutinin (WGA). The inhibitory ability is dependent on the presence of the spacer and on the shape and rigidity of the calixarene skeleton. PMID- 15524434 TI - Counterion-induced translational isomerism in a bistable [2]rotaxane. AB - Translational isomerization can be induced by changing the anions associated with a bistable rotaxane in which the tetracationic cyclophane (blue box), cyclobis(paraquat-p-phenylene), encircles a dumbbell component containing bispyrrolotetrathiafulvalene (green) and a dioxynaphthalene (red) recognition sites. The rotaxane was isolated as both its hexafluorophosphate and tris(tetrachlorobenzenediolato)phosphate(v) (TRISPHAT(-)) salts. Photophysical measurements and NMR spectroscopy carried out in acetone (CD(3)COCD(3)) and acetonitrile (CD(3)CN) solutions reveal that the much larger TRISPHAT(-) anion favors predominantly the encirclement of the green site by the blue box. PMID- 15524435 TI - New platforms integrating ethynyl-grafted modules for organogels and mesomorphic superstructures. AB - A methyldiacylaminophenyl core substituted with gallic derivatives and alkynyl functions has been synthesized from 2,6-diamino-4-iodotoluene. By heating the iodo and ethynyl molecules, typical columnar mesophases are observed and the C(12) synthons bearing a protected terminal alkyne group gelified acetone, via the formation of interlocked fibers which are promoted by intermolecular hydrogen bonding. PMID- 15524436 TI - Sonogashira reaction on quinolizium cations. AB - The four isomeric bromoquinolizium bromides reacted with aryl- and heteroarylacetylenes under Sonogashira conditions. The reactions proceed with moderate-to-high yields to afford aryl- and heteroarylethynyl quinolizium cations. This is the first reported example of the Sonogashira reaction on heteroaromatic cations, and it allowed easy access to potential pi-donor pi acceptor systems bearing cationic units. PMID- 15524437 TI - Radical cation of dibenzothiophene fully annelated with bicyclo[2.2.2]octene units: X-ray crystal structure and electronic properties. AB - New dibenzothiophene 2 fully annelated with bicyclo[2.2.2]octene units was synthesized and oxidized to stable radical cation salt 2(*+)SbCl(6)(-), whose structure was determined by X-ray crystallography. Although the intrinsic electronic structure of 2(*+) is predicted to be close to structure A, an interaction with the counteranion makes structure B contribute significantly. A part of the salt 2(*+)SbCl(6)(-) underwent rearrangement to arenium ion 6(+), whose structure was also clarified by X-ray crystallography. PMID- 15524438 TI - Mild and high-yielding syntheses of diethyl phosphoramidate-stoppered [2]rotaxanes. AB - The mild and efficient reaction between triethyl phosphite and benzylic azides allows us not only to construct rotaxanes in high yield from dibenzo[24]crown-8 (DB24C8) and dibenzylammonium (DBA(+))-derived threads but also to incorporate di(p-toluidine)[24]crown-8, which binds DBA(+) ions much more weakly than does DB24C8, into a corresponding [2]rotaxane. PMID- 15524439 TI - Substituted epoxides by lithiation of terminal epoxides. AB - Diamine-ligand-assisted direct hydrogen-lithium exchange allows the generation of nonstabilized (H-substituted) oxiranyllithium species and their subsequent trapping by Bu(3)SnCl and carbonyl-based electrophiles. This reaction provides a new concise route to alpha,beta-epoxystannanes and substituted epoxides. PMID- 15524440 TI - Lithiation and reactions of stilbene oxides: synthetic utility. AB - The lithiation of trans- and cis-stilbene oxides (+/-)-1 and 8 has been investigated. While with 8, lithiation occurred exclusively at the benzylic position, with the trans isomer (+/-)-1, ortho-lithiation competed with alpha lithiation depending upon the experimental conditions. The configurational stability of the alpha-lithiated cis- and trans-stilbene oxides (+/-)-2 and (+/-) 9, respectively, was proved as well as that of scalemic stilbene oxide (R,R)-2. PMID- 15524441 TI - Synthesis of the Lewis a trisaccharide based on an anomeric silyl fluorous tag. AB - The synthesis of the trisaccharide Lewis a was performed using an anomeric fluorous silyl protective group. This methodology allowed us to fully characterize each product (NMR, MS) and monitor each synthetic step (TLC). Although the product purifications could be performed by fluorous-solid-phase extraction (F-SPE) technology, standard chromatography could be used to effect purification if necessary. Trichloroethoxy carbonyl (Troc) protection of the amino group of the glucosamine moiety was found essential to allow protecting group manipulation of the fluorous protected sugar. PMID- 15524442 TI - Anodic coupling reactions: the use of N,O-ketene acetal coupling partners. AB - Intramolecular anodic olefin coupling reactions utilizing N,O-ketene acetals have been studied. Coupling reactions with both enol ether and allylsilane terminating groups were examined. The reactions involving the coupling of the N,O-ketene acetals with allylsilane groups were found to be much more efficient than corresponding reactions utilizing dithioketene acetal groups and allylsilanes. The reactions were also more efficient than the intramolecular coupling reactions between enol ethers and allylsilanes studied earlier. PMID- 15524443 TI - Sialated diazeniumdiolate: a new sialidase-activated nitric oxide donor. AB - A new sialated diazeniumdiolate has been synthesized, and the glycosylation product was exclusively an alpha anomer. This new nitric oxide donor exhibited significantly improved stability as compared to its parent diazeniumdiolate salts, and it could be efficiently hydrolyzed by neuraminidase to release nitric oxide with a K(m) of 0.14 mM. The sialic acid-NO conjugate would be a valuable prodrug that targets NO to influenza viruses. PMID- 15524444 TI - Investigations of the scope and mechanism of the tandem hydroesterification/lactonization reaction. AB - Heating allylic and homoallylic alcohols and 2-pyridylmethyl formate in the presence of Ru(3)(CO)(12) initiates a tandem sequence of hydroesterification and lactonization. Mechanistic studies suggest that regioselectivity and overall reaction efficiency are governed by the relative rates of reductive elimination and beta-hydride elimination for the alkylruthenium intermediates. PMID- 15524445 TI - One-pot chemo-, regio-, and stereoselective double-differential glycosidation mediated by lanthanide triflates. AB - Nuanced activation of n-pentenyl, thioglycoside, and trichloroacetimidate donors by lanthanide salts coupled with donor/acceptor matching can simplify oligosaccharide assembly. Thus, a one-pot, double-differential glycosidation process can be designed, in which an n-pentenyl acceptor-diol is chemo- and regioselectively glycosidated by using an n-pentenyl ortho ester under the agency of Yb(OTf)(3)/NIS followed by in situ addition of a 2-O-acylated trichloroacetimidate or ethyl thioglycoside to effect stereoselective glycosidation at the remaining OH. PMID- 15524446 TI - Stereoselective preparation of functionalized acyclic alkenylmagnesium reagents using i-PrMgCl.LiCl. AB - Acyclic functionalized alkenyl iodides are converted with high stereoselectivity to the corresponding functionalized alkenylmagnesium derivatives by the reaction with i-PrMgCl.LiCl between -40 and -20 degrees C. Functional groups such as a nitrile, chloride, iodide, and ester are readily tolerated. The conversion of an alkenyl iodide bearing a keto group to the corresponding silylated cyanohydrin allows preparation of the corresponding Grignard reagent affording, after acylation and deprotection, unsaturated 1,4-diketones. PMID- 15524447 TI - Tuning stability of coiled-coil heterotrimers by selection of steric matching partners. AB - Coiled-coil trimers of differing stability have been designed by judicious choice of hydrophobic core side chain identity. Steric matching of two alanines with one large side chain at three core positions affords stable heterotrimers. Use of either cyclohexylalanine (Cha) or phenylalanine (Phe) as the large side chain is effective, and unfolding free energies can be varied by over 3 kcal/mol by mixing Cha- and Phe-containing subunits. PMID- 15524448 TI - A microwave-assisted click chemistry synthesis of 1,4-disubstituted 1,2,3 triazoles via a copper(I)-catalyzed three-component reaction. AB - A microwave-assisted three-component reaction was used to prepare a series of 1,4 disubstituted-1,2,3-triazoles from corresponding alkyl halides, sodium azide, and alkynes. This procedure eliminates the need to handle organic azides, as they are generated in situ, making this already powerful click process even more user friendly and safe. PMID- 15524449 TI - A new hydrogen-bonding motif for chiral recognition in the diastereomeric salts of racemic 1-phenylethylamine derivatives with enantiopure O-ethyl phenylphosphonothioic acid. AB - An enantiopure phosphonothioic acid showed a unique and superior chiral recognition ability, arising from its P-stereogenicity, for racemic 1 phenylethylamine derivatives through diastereomeric crystallization. Spherical molecular clusters, associated by hydrogen bonds and CH/pi interactions, aggregated with high symmetry in the less-soluble diastereomeric salts. PMID- 15524450 TI - Stereochemical control of the Passerini reaction. AB - A catalytic asymmetric Passerini reaction using tridentate indan (pybox) Cu(II) Lewis acid complex 4 with substrates capable of bidentate coordination has been achieved. The reaction occurs via ligand-accelerated catalysis. PMID- 15524452 TI - A new strategy to produce beta-peptides: use of alicyclic beta-lactams. AB - On p-methylbenzhydrylamine (MBHA) resin, by means of t-Boc chemistry, several tetrapeptides (H-Ala-ACXC-Ala-Gly-NH(2)) containing cyclic beta-amino acid units were prepared. These units were introduced into the growing peptide chain by using Boc-protected beta-lactams with KCN as catalyst in DMF. The method was applicable for both racemic and enantiomeric beta-lactams. PMID- 15524453 TI - Preparation of 5-hydroxy-1-alkenyboronates from 1-alkynylboronates, Cp2ZrCl2/2EtMgBr, and aldehydes. AB - 1-Alkynylboronates form five-membered zirconacycles with Cp(2)ZrCl(2)/2-EtMgBr as indicated by deuterium labeling. The zirconacycles add aldehydes to form seven membered zirconacycles. Hydrolysis of the latter provides 5-hydroxy-1 alkenylboronates in fair to good isolated yields. Both aliphatic and aromatic aldehydes undergo insertion. PMID- 15524451 TI - Regioselectivity control in a ruthenium-catalyzed cycloisomerization of diyne ols. AB - The ruthenium-catalyzed cycloisomerization of diynes containing one silyl alkyne and one propargyl alcohol yields 2-silyl-[6H]-pyrans instead of the expected unsaturated acylsilanes except when additional conjugation of a aromatic ring is present at the delta-position. Under certain conditions, a facile ruthenium catalyzed isomerization of the product takes place as well. This regioselectivity of the cyclization can be controlled by the choice of solvent system. DFT calculations confirm the expected greater stability of the silyl-pyrans relative to the acylsilanes. PMID- 15524454 TI - Facile preparation of an orthogonally protected, pH-sensitive, bioconjugate linker for therapeutic applications. AB - We describe the facile, three-step synthesis of an orthogonally protected, pH sensitive linker (8), based on maleic acid, and report its application to the preparation of a pH-sensitive phospholipid (20) for potential use in drug and gene delivery. In addition, we highlight the benefits of our linker over the use of the commercially available cis-aconitic anhydride (4). PMID- 15524455 TI - A concise synthesis of spirotryprostatin A. AB - The preparation of two new synthons, 2,5- and 2,6-dibromotryptophan esters, and their use in diastereoselective intramolecular N-acyliminium ion spirocyclization methodology for the rapid construction of spirotryprostatin A and analogues are described. PMID- 15524456 TI - Site-selective monotitanation of dialkynylpyridines and its application for preparation of highly fluorescent pi-conjugated oligomers. AB - Reaction of Ti(O-i-Pr)(4)/2i-PrMgCl reagent with 2,n bis[(trimethylsilyl)ethynyl]pyridines, where n is 3, 4, 5, and 6, or with 3,4 bis[(trimethylsilyl)ethynyl]pyridines, proceeded with excellent site-selectivity to afford the corresponding monotitanated complex. Synthetic application of the reaction was demonstrated by an efficient preparation of pi-conjugated oligomers having pyridine and enyne units alternately, which possess intense blue fluorescence emission. PMID- 15524457 TI - Synthesis of 2',3'-dideoxy-6',6'-difluorocarbocyclic nucleosides. AB - 2',3'-Dideoxy-6',6'-difluorouracils, a novel series of gem-difluoromethylenated carbocyclic nucleosides, were synthesized from (Z)-but-2-ene-1,4-diol in 14 steps. A notable step was the construction of the carbocyclic ring via ring closing metathesis and the incorporation of gem-difluoromethylene group by way of silicon-induced Reformatskii-Claisen reaction of chlorodifluoroacetic ester 3. PMID- 15524458 TI - Toward the total synthesis of phorboxazole B: an efficient synthesis of the C20 C46 segment. AB - An efficient synthesis of the C20-C46 segment of phorboxazole B is described. The key steps involved Hg(OAc)(2)/I(2)-induced cyclization to construct the cis tetrahydropyran moiety, the coupling of the metalated 2-methyloxazole 7 with lactone 6, and Julia olefination to furnish the conjugated diene moiety. PMID- 15524459 TI - Kinetic control in noncovalent synthesis: regioselective ligand exchange into a hydrogen bonded assembly. AB - This paper illustrates the use of a kinetically controlled exchange reaction to effect regioselective modification of a hydrogen-bonded assembly. Both the bound anion and cation can control the exchange of ligand into the different layers of a synthetic G-quadruplex. PMID- 15524460 TI - A potentially divergent and rapid route to analogues of deoxycyclitols, pentopyranoses, 6-deoxyhexoses, and hexoses. AB - Direct precursors to analogues of pentopyranoses, 6-deoxyhexoses, and hexoses, in which a CF(2) center replaces the pyranose oxygen, have been synthesized rapidly from trifluoroethanol. A simple scaleable allylation reaction delivers ethers which undergo dehydrofluorination/metalation, followed by addition to either acrolein or cinnamaldehyde, to afford allylic alcohols. Fluorine-assisted [3,3] rearrangement followed by reduction with sodium borohydride delivers diols, which undergo RCM smoothly to afford cyclohexene diols. PMID- 15524461 TI - DFT study of pericyclic reaction cascades in the synthesis of antibiotic TAN 1085. AB - DFT calculations show that aromatic and bis-methoxy substituent effects in a synthetic precursor of TAN-1085 strongly favor a [1,7] sigmatropic hydrogen shift over the 6 pi electrocyclic pathway, rendering the latter unfavorable in synthesis. PMID- 15524462 TI - Sugars within a hydrophobic scaffold: glycodendrimers from polyphenylenes. AB - A new glycodendrimer type has been introduced that is designed on the basis of shape-persistent polyphenylene dendrimers. The sugar installation occurs not only on the dendrimer surface but also within the hydrophobic internal scaffold. The synthesis has been accomplished via both convergent and divergent routes by employing the Schmidt glycosylation and the Diels-Alder reaction. This new glycodendrimer has been found to exhibit water-solubility, while conserving hydrophobicity of the interior environment despite the incorporation of sugars. PMID- 15524463 TI - Selective synthesis of alpha,beta-unsaturated ketones by dibutyltin dimethoxide catalyzed condensation of aldehydes with alkenyl trichloroacetates. AB - Various alpha,beta-unsaturated ketones were stereoselectively synthesized in high yields up to 94% by a condensation reaction between alkenyl trichloroacetates and aldehydes using dibutyltin dimethoxide as a catalyst in the presence of methanol. This process is superior to the classical Claisen-Schmidt condensation with respect to mildness of the base catalyst and product selectivity. PMID- 15524464 TI - Synthesis of alpha-fluorosulfonamides by electrophilic fluorination. AB - Alpha-fluorosulfonamides were prepared by electrophilic fluorination of tertiary sulfonamides using N-fluorobenzenesulfonimide as fluorinating agent and utilizing the dimethoxybenzyl group (DMB) as a new sulfonamide protecting group. Removal of the DMB group with TFA/CH(2)Cl(2) gave primary and secondary alpha fluorosulfonamides. PMID- 15524465 TI - Experimental support for planar pseudopericyclic transition states in thermal cheletropic decarbonylations. AB - Low-temperature crystal structures of three pyrrolediones (3-5) and a furandione (9) were obtained and compared to structurally related compounds that cannot undergo decarbonylation. Systematic trends in bond lengths and angles are consistent with distortions along the reaction coordinate, in accord with the structure correlation principle of Dunitz. Since the pyrroledione and furandione rings in 3-5 and 9 are planar, these ground-state geometries prefigure the calculated planar, pseudopericyclic transition states. PMID- 15524466 TI - Solvent-equilibrated homoadamantyl chloride ion pairs from chloroformate or oxachlorocarbene fragmentations. AB - Fragmentations of 3-homoadamantyl chloroformate and 3 homoadamantyloxychlorocarbene produce identical ion pairs as product-determining intermediates. PMID- 15524467 TI - Potassium hydroxide-mediated novel rearrangement of 2-alkyl-sulfonyl-2 arylsulfonyl-1-phenylethanones to 1-aryl-2 (arylsulfonylmethanesulfonyl)ethanones. AB - Treatment of a solution of a mixture of 1-aryl-2-arylsulfonylethanones 9 and alkylsulfonyl chlorides (1.5-2.0 equiv) in THF at 0 degree C with potassium hydroxide (8 equiv) for 10 min gave a rearrangement product, i.e., 1-aryl-2 (arylsulfonylmethanesulfonyl)ethanones 8, in excellent yields. Regiospecific methyl- and ethylation at the methylene carbon sandwiched between two sulfonyl groups of 8 could be achieved by the reactions of 7i-j with LDA (1 equiv) in THF at room temperature, respectively. PMID- 15524468 TI - Efficient synthesis of tris(4-imidazolyl)methanol derivatives. AB - Biomimetic tris(4-imidazolyl)carbinol derivatives are prepared from imidazole in a short, high-yielding sequence via sulfonamide 1, which is converted to the 2 silylated carbinol 2 by one-pot, sequential 2-functionalization and then 4(5) functionalization. Alcohol 2 can be transformed either to the parent carbinol 3 or to a desilylated sulfonamide derivative 4. The tripodal alcohol 3 is a convenient precursor to ethers by solvolysis and to metal complexes, as illustrated by the preparation of a bis-tripod complex with iron(III). PMID- 15524469 TI - Oxidative rearrangement of cyclic tertiary allylic alcohols with IBX in DMSO. AB - A practical and environmentally friendly method for oxidative rearrangement of five- and six-membered cyclic tertiary allylic alcohols to beta-disubstituted alpha,beta-unsaturated ketones by the IBX/DMSO reagent system is described. Several conventional protecting groups (e.g., Ac, MOM, and TBDPS) are compatible under the reaction conditions prescribed. PMID- 15524470 TI - Stereoselective [3,3]-sigmatropic rearrangement promoted by the metal [1,3]-shift of binuclear Fischer carbene complexes. AB - Reaction of chiral homobinuclear Fischer chromium carbene complexes with allyl alcohol in the presence of NaH and the following oxidative demetalation gave alpha-allyl esters in up to 97% ee via [3,3]-sigmatropic rearrangement reaction promoted by the metal 1,3-shift. On the other hand, chiral heterobinuclear tungsten carbene complexes with arene chromium complexes afforded alpha-allyl beta-hydroxy esters as a major product in up to 92/8 dr by the same reaction sequence. PMID- 15524471 TI - A flexible approach toward trans-fused polycyclic tetrahydropyrans. A synthesis of prymnesin and yessotoxin units. AB - Ru-catalyzed cycloisomerization and oxidative cyclization of bis-homopropargylic alcohols provide a rapid iterative approach to structural units of the ladder toxins. PMID- 15524472 TI - Nucleophilic difluoromethylation of primary alkyl halides using difluoromethyl phenyl sulfone as a difluoromethyl anion equivalent. AB - A facile and efficient nucleophilic difluoromethylation of primary alkyl halides has been disclosed through a novel nucleophilic substitution-reductive desulfonylation strategy, using difluoromethyl phenyl sulfone as a difluoromethyl anion ("CF(2)H(-)") equivalent. PMID- 15524473 TI - Axial and equatorial cyclohexylacyl and tetrahydropyranyl-2-acyl radicals. An experimental and theoretical study. AB - Axial and equatorial cyclohexylacyl and tetrahydropyranyl-2-acyl radicals gave distinct EPR spectra thanks to surprisingly large beta-hydrogen atom hyperfine splittings that enabled them to be characterized and monitored. DFT computations indicated that the axial species (X = CH(2)) had a higher barrier to rotation about the (O)C(alpha)-C(beta) bond. The computed difference Delta H degrees for the axial and equatorial radicals (R = H, X = CH(2)) was 0.8 kcal mol(-)(1). PMID- 15524474 TI - Efficient synthesis of 1-adamantanecarboxaldehyde by the GaCl3-mediated carbonylation of adamantane under mild reaction conditions. AB - The reaction of adamantane (1) with CO (1 atm) in 1,2-dichloroethane at room temperature in the presence of GaCl(3) results in formylation to give 1 adamantanecarboxaldehyde (2) in up to 84% yield. PMID- 15524475 TI - Maoecrystal V, cytotoxic diterpenoid with a novel C19 skeleton from Isodon eriocalyx (Dunn.) hara. AB - Maoecrystal V (1), a novel C(19) diterpenoid possessing a unique 6,7-seco-6-nor 15(8-->9)-abeo-5,8-epoxy-ent-kaurane skeleton, was isolated from the leaves of a Chinese medicinal herb, Isodon eriocalyx. Its structure was determined by comprehensive NMR and MS spectroscopic analysis and confirmed by single-crystal X ray diffraction study. Compound 1 showed remarkable inhibitory activity toward HeLa cells with IC(50) = 0.02 microg/mL (cis-platin: IC(50) = 0.99 microg/mL). PMID- 15524476 TI - Requirements for selective hydrophobic acceleration in the reduction of ketones. AB - Reductions of various quaternized hydrophobic beta-keto amines were performed in water and in methanol using borohydride anions carrying hydrophobic groups. The most important requirement of the substrate to permit hydrophobically accelerated selective reductions is the ability of the hydrophobic group of the substrate and its attached keto group to attain a coplanar relationship. Some derivatives of naturally occurring steroid diones have also been employed as substrates to probe the mechanism and utility of these hydrophobically accelerated selective reductions further. PMID- 15524477 TI - Asymmetric synthesis of the tetrahydropyran ring, C32-C38 fragment, of phorboxazoles. AB - The asymmetric synthesis of a model aldehyde (2R,6R)-2 and the C32-C38 fragment of phorboxazoles, (2R,4R,6R)-1, is described using a sulfoxide as chiral auxiliary. Key advances include the stereoselective reductions of beta-keto- or beta,gamma-diketosulfoxides, the acid-catalyzed cyclization of enantiopure sulfinyl hydroxy ketone precursors to the tetrahydropyran ring, and the Pummerer reaction on the pendant sulfoxide to create the formyl group. PMID- 15524478 TI - Synthesis of silyl aziridines and alpha-amino acylsilanes with silyldibromomethyllithium. AB - The reaction of silyldibromomethyllithium with aromatic imines provides alpha amino acylsilanes via a bromo aziridine intermediate upon quenching the reaction with water. Alternatively, treatment of the bromo aziridine intermediate with various Grignard reagents or lithium aluminum hydride permits the nucleophilic displacement of the halogen to furnish substituted silyl aziridines. PMID- 15524479 TI - Three-component, stereoselective palladium-catalyzed synthesis of functionalized bicyclopentanoids. AB - 1,5-Cyclooctadiene can be stereoselectively transformed into a substituted bicyclo[3.3.0]octane ring system under palladium catalysis with concomitant formation of three carbon-carbon bonds. Reaction with an aryl iodide or triflate and malonate gives an exo-endo product, while the reaction with a malonate in the presence of oxygen affords a bis-endo adduct. PMID- 15524480 TI - Rh(II)-catalyzed enantioselective cyclopropanation of olefins with dimethyl malonate via in situ generated phenyliodonium ylide. AB - Olefins are cyclopropanated with dimethyl malonate (1a) iodosylbenzene (PhI=O) and a Rh(II) carboxylate catalyst via an in situ generated phenyliodonium ylide (1c). Enantioselectivities of up to 90% for 4-bromostyrene and 98% for pent-1-ene have been observed with (S)-N-4-bromo-1,8-naphthanoyl-tert-leucine (4c) as the chiral ligand. The same catalyst was effective for olefin cyclopropanation with Meldrum's acid, giving cyclopropanes with 96% (with styrene) and 87% ee (with pent-1-ene), respectively. PMID- 15524481 TI - Use of N-N bond stereodynamics in ring-closing metathesis to form medium-sized rings and macrocycles. AB - A unique strategy based on double ring-closing metathesis for the formation of a 14-membered macrocyclic enamide has been developed. This strategy hinges upon the well-known stereodynamic and conformational behavior of N-substituted diacylhydrazines, which promotes an effective ring-closing metathesis of hydrazine-derived dienes and enynes to form 8- to 14-membered rings. PMID- 15524482 TI - Synthesis and structure of a helical diindenophenanthrene with four congested phenyl substituents as a molecular spiral staircase. AB - Treatment of 9 with potassium tert-butoxide produced 10 having a helical twist in a single operation. The X-ray structure of 10 shows that the four phenyl substituents are essentially parallel to one another but are virtually perpendicular to the helical axis of the diindenophenanthrene ring system. Viewing from the direction perpendicular to the helical axis, the structure of 10 resembles that of a segment of a spiral staircase having four parallel steps. PMID- 15524483 TI - Rhodium-catalyzed novel trifluoromethylation at the alpha-position of alpha,beta unsaturated ketones. AB - Treatment of alpha,beta-unsaturated ketones with CF(3)I in the presence of Et(2)Zn and RhCl(PPh(3))(3) gave novel alpha-trifluoromethylation products in good yields. Hydrogen transfer from the ethyl group on the rhodium complex to the beta-position of the enone seems to play an important role in this reaction. PMID- 15524484 TI - Catalytic additions of acylsilanes to imines: an acyl anion strategy for the direct synthesis of alpha-amino ketones. AB - The addition of acylsilanes to imines catalyzed by neutral carbenes (or zwitterions) generated in situ from readily available thiazolium salts is described. The general reaction successfully utilizes acylsilanes as carbonyl anion precursors and is tolerant of a range of structural diversity on the acylsilane or imine electrophile. The overall reaction utilizes easily available precursors and directly accesses protected alpha-amino ketones in the correct oxidation state. PMID- 15524485 TI - Formation of functionalized carbocycles via base-promoted ring opening/Brook rearrangement/allylic alkylation of gamma-silyl-beta,gamma-epoxybutanenitrile followed by nitrile anion cyclization with bis-electrophiles. AB - Reaction of gamma-silyl-beta,gamma-epoxybutanenitrile with a base generates an alpha-nitrile carbanion derivative of 4-siloxybut-3-enenitrile, which undergoes reaction with bis-electrophiles such as 1,omega-dihaloalkanes, omega-bromo alpha,beta-unsaturated esters, and bisenoates to provide highly functionalized carbocycles. PMID- 15524486 TI - Diastereoselective phenol para-alkylation: access to a cross-conjugated cyclohexadienone en route to resiniferatoxin. AB - We document a route for the synthesis of a densely functionalized spiro-fused 2,5 cyclohexadienone as an intermediate for the synthesis of resineferatoxin. The strategy is based on an unprecedented diastereoselective, intramolecular phenol para-alkylation to a cross-conjugated cyclohexadienone. In the course of these synthetic studies we developed rapid access to a chiral nitrile possessing a quaternary stereocenter and disclose an unusual acetal rearrangement from a dioxane, which favors the corresponding dioxepane. PMID- 15524487 TI - Highly diastereo- and enantioselective reagents for aldehyde crotylation. AB - Two new, crystalline solid, storable, and highly enantioselective reagents for aldehyde crotylation have been developed. Both (cis and trans) crotylsilane reagents are easily prepared in bulk, require trivial reaction conditions, and provide the homoallylic alcohol products with near diastereo- and enantiospecificity in many cases. PMID- 15524488 TI - Palladium-catalyzed cross-coupling of aryl triethylammonium bis(catechol) silicates with aryl bromides using microwave irradiation. AB - The scope of the palladium-catalyzed cross-coupling reaction of aryl bis(catechol) silicates has been extended to include the coupling of aryl bromides by employing microwave irradiation. This new set of coupling conditions is tolerant of electron-rich and -deficient aryl bromides. In addition, a variety of substituted aryl bis(catechol) silicates have been successfully cross-coupled. PMID- 15524489 TI - Total synthesis of callipeltoside A. AB - A convergent total synthesis of cytotoxic marine macrolide callipeltoside A is described. The synthesis highlights two stereoselective [4 + 2] annulations for the preparation of associated pyran rings. PMID- 15524490 TI - Challenge toward structural complexity using asymmetric catalysis: target oriented development of catalytic enantioselective Diels-Alder reaction. AB - A new method for the catalytic enantioselective Diels-Alder reaction using polysubstituted silyl enol ethers as dienes is described. High enantioselectivity (up to 92% ee) was produced using a catalyst generated from FeBr(3) and AgSbF(6) in a 1:2 ratio and aryl-pybox (aryl = Ph or p-ethoxyphenyl). This reaction should facilitate the enantioselective synthesis of polycyclic acylphloroglucinols such as hyperforin or garsubellin A, which are currently of interest from synthetic and medicinal points of view. PMID- 15524491 TI - Gold catalysis: mild conditions for the synthesis of oxazoles from N propargylcarboxamides and mechanistic aspects. AB - 2,5-Disubstituted oxazoles are synthesized from the corresponding propargylcarboxamides under mild reaction conditions via homogeneous catalysis by AuCl(3). While monitoring the conversion via (1)H NMR spectroscopy, an intermediate 5-methylene-4,5-dihydrooxazole can be observed and accumulated up to 95%, being the first direct and catalytic preparative access to such alkylidene oxazolines. The intermediate was fully characterized and can be trapped at -25 degrees C for several weeks. Deuteration experiments show a stereospecific mode of the two first steps of the reaction. PMID- 15524493 TI - Cost-effectiveness analysis of extended-release formulations of oxybutynin and tolterodine for the management of urge incontinence. AB - INTRODUCTION: Oxybutynin and tolterodine are two drugs widely used for the management of overactive bladder and urge urinary incontinence. The once-daily, extended-release formulations benefit from being well tolerated and efficacious. However, their costs, compared with generic immediate-release (IR) oxybutynin, are significantly greater. This study compared the cost effectiveness of oxybutynin extended-release (Oxy-XL), tolterodine extended-release (Tol-ER), tolterodine immediate-release (Tol-IR) and oxybutynin immediate-release (Oxy-IR). STUDY DESIGN: A cost-effectiveness model. METHODS: A systematic review that identified appropriate randomised clinical trials provided evidence on efficacy. Empirical models of drug effects (number of incontinent-free weeks) and persistence (proportion of patients still on therapy) were constructed in order to determine clinical effectiveness which was combined with cost data (direct medical costs to the UK NHS, year 2001 values) to calculate the drugs' cost effectiveness from the perspective of the NHS. Univariate sensitivity analyses were conducted to test the robustness of the results. PATIENTS: Hypothetical cohort of patients with urge incontinence associated with overactive bladder. MAIN OUTCOME MEASURES AND RESULTS: The incremental cost per incontinent-free week for Oxy-IR (versus no treatment) ranged from pound sterling 2.58 to pound sterling 16.59. Oxy-XL and Tol-ER were more effective than Oxy-IR but at additional costs per incontinent-free week. Tol-IR did not appear to be a cost effective option as it was less effective and more costly than the extended release formulations. Uncertainty surrounding the health and cost consequences of early discontinuation affected these results, although the model results were robust to parameter uncertainty. CONCLUSION: Oxy-IR, Oxy-XL and Tol-ER appear to be cost-effective options for the management of urge incontinence from the NHS perspective. A decision among the treatments depends on the acceptable cost per additional incontinent-free week. PMID- 15524492 TI - Clinical and economic impact of epoetins in cancer care. AB - Anaemia develops in most patients undergoing cancer therapy and invariably induces fatigue, which is a major determinant of QOL. Blood transfusions are reserved for patients with severe anaemia, since blood is a scarce resource and provides a short-lived benefit. Epoetins are recombinant proteins capable of alleviating therapy-related anaemia in 40-60% of cancer patients. The number of patients needed to be treated with epoetins to avoid the transfusion of one unit of blood ranges from 2.6 to 5.2; however, the absolute risk reduction depends on patients' characteristics and dose-escalation. The ratio between acquisition costs of epoetins and blood transfusion requirement is very high; thus, many thousands of dollars needs to be spent on epoetins to save 1 blood unit. Despite this, epoetins have been widely adopted by industrialised countries, where cancer patients are about 2% of the total population. The resulting budget impact of epoetins can be calculated at about 10% of the overall direct cost for cancer care, and it is expected to continue growing by about 20% each year, due to the expanding cancer population and the intensification of cancer therapies. The economic burden of epoetins needs to be weighed against the improvement of patients' QOL and society's willingness to pay for a non-life-saving therapy. All published economic evaluations of epoetins invariably report that this supportive therapy is not cost effective. Society should be made aware of the opportunity cost of treatments and should be allowed to elicit preferences for healthcare interventions and prioritisation criteria. In the near future we expect that a wider range of epoetins, drug patent expiry, a more appropriate patient selection criteria and an improved dosage schedule may help increase the efficiency of cancer-related anaemia management. PMID- 15524494 TI - Use and cost of antiretrovirals in France 1995-2000: an analysis based on the Medical Dossier on Human Immunodeficiency (release 2) database. AB - OBJECTIVE: To assess the temporal relationship between the uptake of more aggressive antiretroviral (ARV) therapy and the use and cost of hospital treatment for HIV-infected patients in France from 1995 to 2000 from a hospital perspective. METHODS: Analysis of evolutions in health status (CD4+ cell count, transition to AIDS and mortality) and components of hospital treatment (inpatient hospitalisation, outpatient consultations, day hospital stays and hospital ARV prescriptions) based on a sample of 2203 patients in 30 French hospitals, using data from the French Medical Dossier on Human Immunodeficiency (release 2) database. Health status and healthcare consumption were measured per patient and per semester from 1995 to 2000 inclusive, with unit costs applied to resource data (1999 values). Results were stratified for gravity of illness. RESULTS: Between 1995 and 2000, there was an overall improvement in health status for HIV infected patients. The mortality rate dropped by 82% and the incidence of cases of transition to AIDS fell significantly. Over the same period, the proportion of patients on ARV treatment grew from 69.5% to 97%, with a large rise in the use of polytherapy. The increase was most notable for patients with CD4+ cell counts above 500 cells/mm3, in whom the ARV prescription rate rose from 29% to 96% by 2000. This brought ARV expenditure from Euro 637 per patient per semester in 1995 up to Euro 2042 in 2000, an increase of 220%. Inpatient hospitalisation rates fell by 60% and average length of stay decreased, bringing down total hospital costs (excluding ARVs) from Euro 5179 per patient in the first semester of 1995 to Euro 2314 in 2000 (a 55.5% reduction). Total expenditure per patient per semester therefore fell from Euro 5817 to Euro 4356, a 25% decrease. CONCLUSION: The wider use of multiple combination ARV therapy in France is temporally related to an improvement in the health status of HIV-infected patients, accompanied by reduced consumption of hospital care. PMID- 15524496 TI - Evolution of wrinkles in hard films on soft substrates. AB - A compressively strained film on a substrate can wrinkle into intricate patterns. This Rapid Communication studies the evolution of the wrinkle patterns. The film is modeled as an elastic nonlinear plate and the substrate a viscoelastic foundation. A spectral method is developed to evolve the nonlinear system. When the initial film strains are isotropic, the wrinkles evolve into a pattern with a motif of zigzag segments, in random orientations. When the initial film strains are anisotropic, the wrinkles evolve to an array of herringbones or stripes. The zigzag segments select a width, a length, and an elbow angle that minimize the total elastic energy. PMID- 15524495 TI - Etanercept: a pharmacoeconomic review of its use in rheumatoid arthritis. AB - Etanercept (Enbrel), which inhibits the activity of tumour necrosis factor-alpha, is indicated in the treatment of patients with active rheumatoid arthritis (RA). A lifetime cost-utility analysis in patients with severe disease-modifying antirheumatic drug (DMARD)-resistant RA in the UK suggested that etanercept is associated with acceptable cost-utility ratios relative to traditional nonbiological DMARDs. In a 12-month cost-utility study in Spain, etanercept was predicted to be dominant over infliximab plus methotrexate in patients with active, refractory RA with regards to the cost per QALY gained and cost per American College of Rheumatology (ACR) 20 response achieved. In short-term cost effectiveness analyses conducted in the US, the cost effectiveness of etanercept relative to other treatments in patients with methotrexate-naive or -resistant RA depends on whether predicted incremental cost-effectiveness ratios of at least USD 41,900 per ACR 20 response or USD 34,800 per ACR 70 weighted response over a 6-month period are considered acceptable (1999 values). The relative efficacy and cost effectiveness of etanercept and other biological DMARDs will be clarified when appropriate data from directly comparative clinical and/or long-term pharmacoeconomic studies become available. Etanercept may prevent or delay disability, which may produce reductions in nondrug costs that could help offset its acquisition cost. PMID- 15524497 TI - Multiphoton switching dynamics of single green fluorescent proteins. AB - Multi-photon driven photo-switching between dark and bright (fluorescent) states of a green fluorescent protein (GFP) mutant is demonstrated. A single-protein investigation shows the existence of two distinct bright states that display sharp two-photon cross-section bands peaked at 780 nm and at 870 nm. Fluorescence of these two species can be independently switched on and off. These results highlight a new photoconversion pathway for photochromic GFPs and can have significant applications in multi-photon confocal microscopy and in optical data storage architectures. PMID- 15524498 TI - Unexpected epidemic thresholds in heterogeneous networks: the role of disease transmission. AB - We reformulate several recent analyses of infection processes on highly heterogeneous networks (e.g., scale-free networks) which conclude that diseases will spread and persist even for vanishingly small transmission probabilities. The results of these latter studies contrast with conventional epidemiological models where there are clear threshold effects, namely, should the transmission probability fall below a critical threshold level the disease is expected to die out. Here we show that epidemic propagation depends equally on the infection scheme as well as the network structure. Connectivity-dependent infection schemes can yield threshold effects even in scale-free networks where they would otherwise be unexpected. PMID- 15524499 TI - Teaching computers to fold proteins. AB - A new general algorithm for optimization of potential functions for protein folding is introduced. It is based upon gradient optimization of the thermodynamic stability of native folds of a training set of proteins with known structure. The iterative update rule contains two thermodynamic averages which are estimated by (generalized ensemble) Monte Carlo. We test the learning algorithm on a Lennard-Jones (LJ) force field with a torsional angle degrees-of freedom and a single-atom side-chain. In a test with 24 peptides of known structure, none folded correctly with the initial potential functions, but two thirds came within 3 A to their native fold after optimizing the potential functions. PMID- 15524500 TI - Intermittency of height fluctuations in stationary state of the Kardar-Parisi Zhang equation with infinitesimal surface tension in 1+1 dimensions. AB - The Kardar-Parisi-Zhang (KPZ) equation with infinitesimal surface tension, dynamically develops sharply connected valley structures within which the height derivative is not continuous. We discuss the intermittency issue in the problem of stationary state forced KPZ equation in 1+1 dimensions. It is proved that the moments of height increments C(a) = <|h(x(1)) - h(x(2))|(a)> behave as |x(1) - x(2)|(xi(a)) with xi(a) = a for length scales |x(1) - x(2)|<0) interact and exchange the quantities asymmetrically by the rule x(') =c [(1-a) x+by] , y(') =d [ax+(1-b) y] (x> or =y) , where (0< or =) a,b (< or =1) and c,d (>0) are interaction parameters. Noninteger power-law tails in the probability distribution function of scaled quantities are analyzed in a similar way as in inelastic Maxwell models. A transcendental equation to determine the growth rate gamma of the processes and the exponent s of the tails is derived formally from moment equations in Fourier space. In the case c=d or a+b=1 (a not = 0, 1) , the first-order moment equation admits a closed form solution and gamma and s are calculated analytically from the transcendental equation. It becomes evident that at c=d , exchange rate b of small quantities is irrelevant to power-law tails. In the case c not = d and a+b not = 1 , a closed form solution of the first-order moment equation cannot be obtained because of asymmetry of interactions. However, the moment equation for a singular term formally forms a closed solution and possibility for the presence of power-law tails is shown. Continuity of the exponent s with respect to parameters a,b,c,d is discussed. Then numerical simulations are carried out and compared with the theory. Good agreement is achieved for both gamma and s. PMID- 15524506 TI - Consistent description of quantum Brownian motors operating at strong friction. AB - A quantum Smoluchowski equation is put forward that consistently describes thermal quantum states. In particular, it notably does not induce a violation of the second law of thermodynamics. This so modified kinetic equation is applied to study analytically directed quantum transport at strong friction in arbitrarily shaped ratchet potentials that are driven by nonthermal two-state noise. Depending on the mutual interplay of quantum tunneling and quantum reflection these quantum corrections can induce both, a sizable enhancement or a suppression of transport. Moreover, the threshold for current reversals becomes markedly shifted due to such quantum fluctuations. PMID- 15524507 TI - Transport and fractionation in periodic potential-energy landscapes. AB - Objects driven through periodically modulated potential-energy landscapes in two dimensions can become locked in to symmetry-selected directions that are independent of the driving force's orientation. We investigate this problem in the overdamped limit, and demonstrate that the crossover from free-flowing to locked-in transport can depend exponentially on an object's size, with this exceptional selectivity emerging from the landscape's periodicity. PMID- 15524508 TI - Density-functional theory for the structures and thermodynamic properties of highly asymmetric electrolyte and neutral component mixtures. AB - Density-functional theory (DFT) is applied to investigate the structural and thermodynamic properties of concentrated electrolyte and neutral component mixtures that are highly asymmetric in terms of both size and charge mimicking a crowded cellular environment. The excess Helmholtz energy functional is derived from a modified fundamental measure theory for the hard-sphere repulsion and a quadratic functional Taylor expansion for the electrostatic interactions. The direct correlation functions are obtained from the analytical solutions of the mean-spherical approximation. In the context of a primitive model where biomacromolecules are represented by neutral or charged hard spheres and the solvent is represented by a continuous dielectric medium, this DFT is able to take into account both the excluded-volume effects and the long-ranged electrostatic interactions quantitatively. The performance of the theoretical method has been tested with Monte Carlo simulation results from this work and from the literature for the pair correlation functions, excess internal energies, and osmotic coefficients for a wide variety of aqueous dispersions of charged and neutral particles. PMID- 15524509 TI - On-lattice coalescence and annihilation of immobile reactants in loopless lattices and beyond. AB - We study the behavior of the chemical reactions A+A-->A+S and A+A-->S+S (where the reactive species A and the inert species S are both assumed to be immobile) embedded on Bethe lattices of arbitrary coordination number z and on a two dimensional (2D) square lattice. For the Bethe lattice case, exact solutions for the coverage in the A species in terms of the initial condition are obtained. In particular, our results hold for the important case of an infinite one dimensional (1D) lattice (z=2). The method is based on an expansion in terms of conditional probabilities which exploits a Markovian property of these systems. Along the same lines, an approximate solution for the case of a 2D square lattice is developed. The effect of dilution in a random initial condition is discussed in detail, both for the lattice coverage and for the spatial distribution of reactants. PMID- 15524510 TI - Localized states on comb lattices. AB - Complex networks and graphs provide a general description of a great variety of inhomogeneous discrete systems. These range from polymers and biomolecules to complex quantum devices, such as arrays of Josephson junctions, microbridges, and quantum wires. We introduce a technique, based on the analysis of the motion of a random walker, that allows us to determine the density of states of a general local Hamiltonian on a graph, when the potential differs from zero on a finite number of sites. We study in detail the case of the comb lattice and we derive an analytic expression for the elements of the resolvent operator of the Hamiltonian, giving its complete spectrum. PMID- 15524511 TI - Random cuts in binary mixtures of spheres. AB - Three-dimensional (3D) disordered media may be studied by performing random planar cuts through the material and trying to go back to 3D geometry from two dimensional information. We have studied from this point of view numerical packings of spheres of two sizes for which the radii of the spheres and the composition of the mixtures have been obtained from the cuts. We have also studied the froths generated by the related Laguerre-Voronoi tessellation of the packings, and their cuts. Stereological relations, in this specific frame, are derived in both cases. PMID- 15524512 TI - Transitions and transport for a spatially periodic stochastic system with locally coupled oscillators. AB - In this paper, with a special model, we investigate the spatially periodic stochastic system with locally coupled oscillators subject to a constant force F. A nonequilibrium second-order phase transition is found when F=0. This phase transition is reentrant when the additive noise is weak. With varying the constant force F, a continuous or discontinuous transition between the states with positive and negative mean fields (mu>0 and mu<0) is observed, which is not a phase transition. The mean field or current sometimes exhibits hysteresis as a function of F. With the variation of the force F, when hysteresis of the mean field or current versus F appears, a nonzero probability current with definite direction will occur at the point F=0. The correlation between the additive and multiplicative noises has an effect on the transitions and the transport. PMID- 15524513 TI - Demixing in simple dipolar mixtures: Integral equation versus density functional results. AB - Using reference hypernetted chain (RHNC) integral equations and density functional theory in the modified mean-field (MMF) approximation we investigate the phase behavior of binary mixtures of dipolar hard spheres. The two species (A and B) differ only in their dipole moments m(A) and m(B), and the central question investigated is under which conditions these asymmetric mixtures can exhibit demixing phase transitions in the fluid phase regime. Results from our two theoretical approaches turn out to strongly differ. Within the RHNC (which we apply to the isotropic high-temperature phase) demixing does indeed occur for dense systems with small interaction parameters Gamma= m(2)(B)/m(2)(A). This result generalizes previously reported observations on demixing in mixtures of dipolar and neutral hard spheres (Gamma=0) to the case of true dipolar hard sphere mixtures. The RHNC approach also indicates that these demixed fluid phases are isotropic at temperatures accessible by the theory, whereas isotropic-to ferroelectric transitions occur only at larger Gamma. The MMF theory, on the other hand, yields a different picture in which demixing occurs in combination with spontaneous ferroelectricity at all Gamma considered. This discrepancy underlines the relevance of correlational effects for the existence of demixing transitions in dipolar systems without dispersive interactions. Indeed, supplementing the dipolar interactions by small, asymmetric amounts of van der Waals-like interactions (and thereby supporting the systems tendency to demix) one finally reaches good agreement between MMF and RHNC results. PMID- 15524514 TI - Symmetric diffusion equations, barodiffusion, and cross-diffusion in concentrated liquid mixtures. AB - In models of diffusion in multicomponent mixtures, the current practice is to derive equations for an isobaric system. The equations are nonsymmetric in relation to the components of the mixture, and the concentration of solvent is assumed to be governed by the conservation of mass instead of its own corresponding diffusion equation. For concentrated mixtures, the solvent component is selected arbitrarily, which makes interpretation of the experimental data dependent on the choice of the interpreter. In this work, we derive a symmetric system of equations, made possible by the introduction of a spontaneously produced secondary pressure gradient. The effect of that pressure gradient is barodiffusion (barophoresis), defined by the force expressed as the secondary pressure gradient multiplied by the molecular volume. The model also considers the cross-diffusion (diffusiophoresis) that results from the hydrodynamic stresses associated with the local concentration-induced pressure gradient in liquid layers surrounding individual molecules. The resulting system of diffusion equations, which contains the secondary pressure gradient and component concentrations, is applied to a binary (nonionic) mixture of benzene and 1,2-dichloroethane. The steady-state system is placed in a uniform force field, and the effect of the secondary pressure gradient on the field-induced migration is discussed. Fluctuation dynamics in a system with no external force field is also discussed. The numerical results predict the establishment of lower concentration gradients compared to standard theory. Also, the predicted concentration dependence in the effective diffusion coefficient measured by dynamic light scattering is different compared to standard theory. Finally, experiments are proposed to further evaluate differences between the new model and the standard approach. PMID- 15524515 TI - Effect of interparticle force on mixing and segregation of dry granular materials. AB - In this paper, the effects of interparticle force on mixing, segregation, and stratification in dry granular materials are investigated. Avalanche segregation, stratification and also segregation in rotating drums are examined. A series of binary mixtures of granular materials is prepared which consists of spherical iron particles and a nonmagnetic material. By placing each mixture in a magnetic field, the induced magnetic interparticle force could be altered and the effects on particle segregation observed. Using this technique, the effects of altering interparticle force on both avalanche and radial segregation are examined. It is found that altering interparticle force could induce mixed materials to segregate and also induce segregating granular materials to mix. We also report a complete reversal of segregation and stratification as interparticle force was increased. These results have important implications for the mixing of cohesive powders. PMID- 15524516 TI - van der Waals normal form for a one-dimensional hydrodynamic model. AB - Phase separation in a fluidized granular system is studied. We consider a one dimensional hydrodynamic model that mimics a two-dimensional fluidized granular system with a vibrating wall and without gravity, which exhibits a phase separation. Close to the critical point, by means of an adiabatic elimination of the temperature, we deduce the van der Waals normal form, which is the equation that describes the slow dynamics of the system and predicts the qualitative behavior in different regions of parameters. This allows us to understand the origin of the effective viscosity and the spatial saturation at the onset of the bifurcation. The hydrodynamic model and van der Waals normal form exhibit a behavior similar to the one observed in molecular dynamics simulations. PMID- 15524517 TI - Slowly sheared dense granular flows: crystallization and nonunique final states. AB - Simultaneous time-resolved measurements of internal structure, granular volume, and boundary shear force are reported for dense granular packing steadily sheared under a fixed normal load. We identify features of the crystallization transition for a deep shear flow, whose height-dependent local mean velocity spans more than five orders of magnitude. This structural change is accompanied by a significant decrease of granular volume and shear force, with a more rapid falloff of particle velocity with depth than occurs in the disordered state. Boundary conditions can have a profound influence on the crystallization of the entire packing. We find that continuously sheared flow can exhibit nonunique final states even under identical boundary conditions; a few cycles of oscillatory pretreatment can initiate states that evolve into either a crystallized or a disordered final state after long-term unidirectional shearing. On the other hand, the disordered state can be stabilized after being sufficiently compacted by unidirectional shear. These experiments raise interesting questions about how prior history is recorded in the internal structure of granular packings, affecting their instantaneous rheology and long-term evolution in response to shear. PMID- 15524518 TI - Segregation in a monolayer of magnetic spheres. AB - Segregation and pattern formation is investigated for binary mixtures of granular magnetic spheres in a vertically vibrated monolayer. The spheres, all of equal mass and size, have a maximum surface magnetic field B induced by encased cylindrical magnetic cores of length l. For binary mixtures of particles with equal l but different B, we find that the particles spontaneously segregate when driven. For fixed vibration frequency, the segregation rate increases roughly linearly with driving acceleration over the amplitudes investigated. For systems of fixed particle number density, the rate of segregation also decreases as the volume fraction of "strong" (high B) particles increases. We find that segregation also occurs in binary mixtures of particles with equal B, but different l. Finally, using a simple model of spheres with dipolar and higher magnetic moments, we show that the observed segregation phenomena occur in conjunction with a decrease in magnetic energy. PMID- 15524519 TI - Analytic study of the three-urn model for separation of sand. AB - We present an analytic study of the three-urn model for separation of sand, which can be regarded as a zero-range process. We solve analytically the master equation and the first-passage problem. We find that the stationary probability distribution obeys the detailed balance and is governed by the free energy. We find that the characteristic lifetime of a cluster diverges algebraically with exponent 1/3 at the limit of stability. We also give a general argument that the scaling behavior is robust with respect to different expressions of the flux. PMID- 15524520 TI - Determination of branch fraction and minimum dimension of mass-fractal aggregates. AB - Particles of micrometer to nanometer size often aggregate to form branched structures. Such materials include metals and metal oxides as well as biological and polymeric materials (considering the persistence length as a primary unit). Characterization of such structures is difficult since they typically display disordered, irregular features in three dimensions. Branched aggregates display two limiting size scales: that of the primary particle, R1 and that of the aggregate, R2. The mass-fractal model is often used to describe such structures where the aggregate mass, z=M2/M1, is related to the aggregate size, r=R2/R1, through a scaling relationship z=alpha r (d(f)), where the lacunarity alpha is close to 1 and may depend on the growth mechanism. Scattering of x rays, light and neutrons yields a direct measure of the mass-fractal dimension since I(q) approximately q(-d(f)) for 1/R2 rho(s), i.e., the liquid expands upon solidification. Then, the growth is contrasted by an increasing pressure, which results in a continuous decrease of the coexistence temperature and the effective supercooling. These phenomena have been simulated in two dimensions through a modified version of the classic phase field model. We observe that for spherical growth the interface temperature reflects almost instantaneously the change of the coexistence temperature. For dendritic growth, we observed a relaxation time for the dendrite tip velocity and the tip radius which is comparable to the characteristic time of the process; however, after the first fast transient, the growth dynamics seems to follow the changing pressure with no appreciable lag. The onset of the morphological instability is slightly anticipated in respect to free growth. PMID- 15524530 TI - Sign of line tension in liquid bridge stability. AB - We apply the stability criterion we recently proposed for a general wetting functional [Phys. Rev. E 68, 012601 (2003)] to find out whether straight liquid bridges can be stable when subject to line tension of either sign. Our main conclusion is that, even when the line tension is negative, a straight liquid bridge can be stable, and so observable, provided that the line tension is not too large in absolute value. PMID- 15524531 TI - Growth rate of a crystal facet of arbitrary size and growth kinetics of vertical nanowires. AB - We present a modification of the Kolmogorov-Johnson-Mehl-Avrami crystallization model to the case of a finite size crystal facet growing layer by layer. A general expression for the facet growth rate is derived that provides an asymptotic matching to the known limit cases of very small and very large facets. The derived expression is applied to the study of the growth kinetics of vertical nanowires in the "vapor-liquid-solid" growth mechanism. The presented model generalizes the Givargizov-Chernov theory of whisker growth, shows why the whiskers grow much faster than the nonactivated surface, and gives the dependence of the growth rate of nanowires on the diameter of drop of liquid alloy and the growth conditions. PMID- 15524532 TI - Adsorption phenomenon of neutral particles and a kinetic equation at the interface. AB - The adsorption phenomenon of neutral particles in a sample having the shape of a slab is theoretically investigated by using a particular form for the kinetic equation at the limiting surfaces. The time evolution of the bulk and surface densities is determined in a closed form by means of a simple expression. A discussion on the characteristic times entering in the problem is reported. Finally, a microscopic model giving rise to a kinetic equation similar to the one used in the analysis is proposed, and the phenomenological parameters determined. The analysis is suitable for the description of the adsorption phenomena of dyes in nematic liquid crystals. PMID- 15524533 TI - Analytic formulation and parametrization of the kinetic potential theory for drizzle formation. AB - The kinetic potential of nucleation theory is extended to describe cloud droplet growth processes that can lead to drizzle formation. In this model drizzle formation is identified as a statistical barrier crossing phenomenon that transforms cloud droplets to much larger drizzle size with a rate dependent on turbulent diffusion, droplet collection efficiency, and properties of the underlying cloud droplet size distribution. Closed-form expressions for the kinetic potential, critical drop volume, barrier height, and both steady-state and transient barrier crossing drizzle rates are obtained in terms of measurable cloud properties. In an analogy with the theory of phase transformation, clouds are classified into two regimes: an activated metastable regime, in which there is a significant barrier and drizzle initiation resembles nucleation, and an unstable regime where kinetics dominates analogous to the spinodal regime of phase transformation. Observational evidence, including the threshold behavior of drizzle formation and the well-known effect that aerosols have on drizzle suppression, is shown to favor drizzle formation under activated conditions (more similar to nucleation than spinodal decomposition) and under transient conditions rather than steady state. These new applications of the kinetic potential theory should lead to more accurate parametrizations of aerosol-cloud interaction and improved algorithms for weather forecasting and climate prediction. PMID- 15524534 TI - Numerical study of discrete models in the class of the nonlinear molecular beam epitaxy equation. AB - We study numerically some discrete growth models belonging to the class of the nonlinear molecular beam epitaxy equation, or the Villain-Lai-Das Sarma (VLDS) equation. The conserved restricted solid-on-solid model (CRSOS) with maximum height differences Delta H(max)=1 and Delta H(max)=2 was analyzed in substrate dimensions d=1 and d=2 . The Das Sarma and Tamborenea (DT) model and a competitive model involving random deposition and CRSOS deposition were studied in d=1. For the CRSOS model with Delta H(max)=1, we obtain the more accurate estimates of scaling exponents in d=1:roughness exponent alpha=0.94+/-0.02 and dynamical exponent z=2.88+/-0.04. These estimates are significantly below the values of one-loop renormalization for the VLDS theory, which confirms Janssen's proposal of the existence of higher-order corrections. The roughness exponent in d=2 is very near the one-loop result alpha=2/3, in agreement with previous works. The moments W(n) of orders n=2 , 3, 4 of the height distribution were calculated for all models, and the skewness S triple bond W3/W(3/2)(2) and the kurtosis Q triple bond W4/W(2)2-3 were estimated. At the steady states, the CRSOS models and the competitive model have nearly the same values of S and Q in d=1, which suggests that these amplitude ratios are universal in the VLDS class. The estimates for the DT model are different, possibly due to their typically long crossover to asymptotic values. Results for the CRSOS models in d=2 also suggest that those quantities are universal. PMID- 15524535 TI - Characterizing the microscopic physics near moving contact lines using dynamic contact angle data. AB - Directly probing the fluid flow and liquid-vapor interface shape in the microscopic immediate vicinity of the moving contact line can only be accomplished in very specific and isolated cases. Yet this physics is critical to macroscopic dynamic wetting. Here we examine the microscopic (or inner) physics of spreading silicone fluids using data of macroscopic dynamic contact angle versus Capillary number Ca=U mu/sigma. This dynamic contact angle is precisely defined so that it can be related back to the microscopic behavior through detailed theory. Our results indicate that the parameters describing the inner region have a detectable dependence on spreading velocity when this velocity exceeds a critical value. This dependence is not scaled (i.e., the data are not collapsed) by Ca, which suggests that an additional time scale must be present in the model of the inner region. PMID- 15524536 TI - Ripening of porous media. AB - We address the surface-tension-driven dynamics of porous media in nearly saturated pore-space solutions. We linearize this dynamics in the reaction limited regime near its fixed points--surfaces of constant mean curvature (CMC surfaces). We prove that the only stable interface for this dynamics is the plane and estimate the time scale for a CMC surface to become unstable. We also discuss the differences between dynamics in open and closed environments, pointing out the unlikelihood that CMC surfaces are ever realized in such environments on any time scale. PMID- 15524537 TI - Collective and local molecular dynamics in the lyotropic mesophases of decylammonium chloride: 1H and 2H NMR study. AB - The collective and individual dynamics of decylammonium chloride (DACl) molecules in water environment were investigated as a function of surfactant concentration and temperature. In the presence of water the DACl forms a variety of self assembled structures, ranging from isotropic micellar systems to lyotropic liquid crystalline phases of hexagonal, nematic, and lamellar types. In order to characterize the complex molecular dynamics that occur in the DACl-water system, we applied 1H and 2H NMR techniques that cover the whole frequency range between 1 kHz and 30 MHz. The slow molecular dynamics were studied by 1H NMR fast-field cycling T1 measurements and pulse-frequency dependence of 2H NMR transverse relaxation time, performed by means of the Carr-Purcell-Meiboom-Gill sequence. We detected a well-expressed contribution of order director fluctuations, i.e., layer undulations, with characteristic omega(-1)(L) frequency dependence of T( 1)(1) in the lamellar phase. Its presence indicates a relatively weak impact of interactions between neighboring DACl layers. The frequency dependence of proton T(-1)(1) in the hexagonal phase exhibits a different type of frequency dispersion, T(-1)(1) approximately omega(-1.32)(L). The increase in the exponent is explained with the quasi-one-dimensional character of fluctuations in elongated cylinders. Further, the T1 and T2 relaxation times of deuterons selectively attached to the C2 and C7 segments of the hydrocarbon chains of DACl were measured at a Larmor frequency of 30.7 MHz, providing quantitative information about local molecular dynamics. PMID- 15524538 TI - Bent-core dopant in a liquid crystal having a reentrant synclinic phase. AB - Small quantities of the bent-core mesogen P-7PIMB were dissolved in an anticlinic liquid crystal consisting of a mixture of left- and right-handed TFMHPOBC, with enantiomer excess X=0.2. The bent-core dopant promotes anticlinic order at higher temperatures, but becomes less effective in suppressing the synclinic phase at the reentrant synclinic transition due to an orientational transition of the dopant within the calamitic TFMHPOBC matrix. Measurements of the anticlinic synclinic electric-field switching threshold as a function of temperature and dopant concentration facilitate a determination of the component of the anticlinic interaction coefficient U that is due to the bent-core dopant. It is found that the value of U per bent-core molecule is much larger than the corresponding value for a pair of TFMHPOBC molecules in adjacent smectic layers. PMID- 15524539 TI - Polarity-directed analog electro-optic switching in a low-polarization chiral smectic liquid crystal with positive dielectric anisotropy. AB - We describe an analog electro-optic (EO) switching mechanism occurring in thin cells filled with a low-polarization ferroelectric liquid crystal mixture with positive dielectric anisotropy. The mixture is composed of an achiral nonpolar smectic-C (Sm-C) host doped with a small amount of a commercially available unichiral compound. The switching mechanism provides analog EO behavior, and thus could be attractive for information display applications. The process is polarization-driven for weak fields, while for higher field strength the dielectric coupling dominates the process. PMID- 15524540 TI - Direct measurement of the permeability of the meniscus bordering a free-standing smectic-A film. AB - A smectic-A free-standing film is always connected by a meniscus to the frame on which it has been stretched. The meniscus acts as a dissipative reservoir and is characterized by its permeability. We propose a method to measure directly this quantity by equilibrating two menisci in correspondence with the same free standing film. The permeability is shown to depend on the film thickness, in full agreement with previous indirect measurements obtained by analyzing the growth dynamics of dislocation loops. An improved model of the meniscus is proposed to interpret all the data. PMID- 15524541 TI - Electrostatic contribution to the persistence length of a semiflexible dipolar chain. AB - We investigate the electrostatic contribution to the persistence length of a semiflexible polymer chain whose segments interact via a screened Debye-Huckel dipolar interaction potential. We derive the expressions for the renormalized persistence length on the level of a 1/D-expansion method already successfully used in other contexts of polyelectrolye physics. We investigate different limiting forms of the renormalized persistence length of the dipolar chain and show that, in, general, it depends less strongly on the screening length than in the context of a monopolar chain. We show that for a dipolar chain the electrostatic persistence length in the same regime of the parameter phase space as the original Odijk-Skolnick-Fixman (OSF) form for a monopolar chain depends logarithmically on the screening length rather than quadratically. This can be understood solely on the basis of a swifter decay of the dipolar interactions with separation compared to the monopolar electrostatic interactions. We comment also on the general contribution of higher multipoles to the electrostatic renormalization of the bending rigidity. PMID- 15524542 TI - Overdamped stress relaxation in buckled rods. AB - We present a comprehensive theoretical analysis of the stress relaxation in a multiply but weakly buckled incompressible rod in a viscous solvent. For the bulk, two interesting parameter regimes of generic self-similar intermediate asymptotics are distinguished, which give rise to approximate and exact power-law solutions, respectively. For the case of open boundary conditions the corresponding nontrivial boundary-layer scenarios are derived by a multiple-scale perturbation ("adiabatic") method. Our results compare well with--and provide the theoretical explanation for--previous results from numerical simulations, and they suggest directions for further fruitful numerical and experimental investigations. PMID- 15524543 TI - Half-soliton interaction of population taxis waves in predator-prey systems with pursuit and evasion. AB - In this paper, we use numerical simulations to demonstrate a half-soliton interaction of waves in a mathematical model of a "prey-predator" system with taxis when of two colliding waves, one annihilates and the other continues to propagate. We show that this effect depends on the "ages" or, equivalently, "widths" of the colliding waves. In two spatial dimensions we demonstrate that the type of interaction, i.e., annihilation, quasisoliton, or half-soliton, depends not only on curvature and width of the colliding waves, but also on the angle of the collision. When conditions of collision are varying in such a way that only a part of a wave survives the collision, then "taxitons," compact pieces of solitary waves, may form, which can exist for a significant time. PMID- 15524544 TI - Quantum models of the mind: are they compatible with environment decoherence? AB - Quantum models of the mind associate consciousness with coherent superposition of states in the brain. Some authors consider consciousness to be the result of a kind of internal quantum measurement process in the brain. In this paper we discuss the ideas of Hameroff-Penrose and Tegmark and their calculation for an estimate of decoherence time. We criticize the Hameroff-Penrose model in the context of a quantum brain model by gravitational collapse orchestrated objective reduction (OOR), assumed by Penrose, and we propose instead that the decoherence process is caused by interaction with the environment. We consider it useful to exploit this possibility because of the growing importance of the decoherence theory in quantum measurement, and also because quantum mechanics can be applied to brain study independently of the Hameroff-Penrose model for mind and consciousness. Our conclusion is that the Hameroff-Penrose model is not compatible with decoherence, but nevertheless the quantum brain can still be considered if we replace gravitational collapse OOR with decoherence. However, our result does not agree with Tegmark's conclusion of refuting not only the Hameroff-Penrose gravitational collapse but also the quantum brain, based on decoherence time calculations in specific cases in the brain. In spite of this fact we also disagree with some points of the response to Tegmark's article given by Hagan, Hameroff, and Tuszynski. PMID- 15524545 TI - Bioinspired laser-operated molecular locomotive. AB - Biomotors kinesin and dynein show us that robust track-walking is possible down to molecular scale. Here I design a laser-powered molecular locomotive that is able to do that on an easily constructed track. The core of the machine is its work cycle that periodically converts optical energy into mechanical work, which is further rectified into processive, directional motion. Thus the molecular locomotive is essentially beyond the famous design of molecular shuttles. Under automated laser operation, the locomotive can move a few mum per second comparable to its biological counterparts. However, this artificial motor is capable of conveniently switchable, dual directional motion in contrast to common unidirectionality of biomotors. The locomotive is also different from the big category of Brownian motors in the sense that move of the locomotive is not a result of biasing pre-existing fluctuations, rather it is directly and decisively driven by optomechanical strokes of the work cycle, generating a pulling force ten times greater than those of biomotors. Being a novel type of molecular motor as well as a powerful molecular engine, this machine will potentially enable automatic, forceful delivery of molecular building blocks with nanometer accuracy. Well within reach of established techniques, its implementation will be a significant advance in nanoscience and nanotechnology. PMID- 15524546 TI - Influence of time-delayed feedback in the firing pattern of thermally sensitive neurons. AB - We explore the dynamics of a Hodgkin-Huxley-type model for thermally sensitive neurons that exhibit intrinsic oscillatory activity. The model is modified to include a feedback loop that is represented by two parameters: the synaptic strength and the transmission delay time. We analyze the dynamics of the neuron depending on the temperature, the synaptic strength, and the delay time. We find parameter regions where the effect of the recurrent connexion is excitatory, inducing spikes or trains of spikes, and regions where it is inhibitory, reducing or eliminating completely the spiking behavior. We characterize the complex interplay of the intrinsic dynamics of the neuron with the recurrent feedback input and a noisy input. PMID- 15524547 TI - Self-organized pattern formation in motor-microtubule mixtures. AB - We model the stable self-organized patterns obtained in the nonequilibrium steady states of mixtures of molecular motors and microtubules. In experiments [Nedelec et al., Nature (London) 389, 305 (1997); Surrey et al., Science 292, 1167 (2001)] performed in a quasi-two-dimensional geometry, microtubules are oriented by complexes of motor proteins. This interaction yields a variety of patterns, including arrangements of asters, vortices, and disordered configurations. We model this system via a two-dimensional vector field describing the local coarse grained microtubule orientation and two scalar density fields associated to molecular motors. These scalar fields describe motors which either attach to and move along microtubules or diffuse freely within the solvent. Transitions between single aster, spiral, and vortex states are obtained as a consequence of confinement, as parameters in our model are varied. For systems in which the effects of confinement can be neglected, we present a map of nonequilibrium steady states, which includes arrangements of asters and vortices separately as well as aster-vortex mixtures and fully disordered states. We calculate the steady state distribution of bound and free motors in aster and vortex configurations of microtubules and compare these to our simulation results, providing qualitative arguments for the stability of different patterns in various regimes of parameter space. We study the role of crowding or "saturation" effects on the density profiles of motors in asters, discussing the role of such effects in stabilizing single asters. We also comment on the implications of our results for experiments. PMID- 15524548 TI - Spatially resolved electronic detection of biopolymers. AB - An integrated array of field-effect transistor structures is used to detect two oppositely charged biopolymers: poly(L-lysine) and DNA. Local deposition of polymer solutions on part of the array induces sizeable variations in the dc current-voltage characteristics of the transistors exposed to the molecular charge. The whole transistor array is measured in the presence of a common electrolyte. Differential signals are studied as a function of electrolyte salt and polymer concentrations. The measurements provide information on the interface electrostatic potentials of the (semiconductor/biopolymer/electrolyte) system and the experimental data are compared to an analytical model which accounts for screening of the adsorbed charge by mobile ions. PMID- 15524549 TI - Motion-contrast computation without directionally selective motion sensors. AB - The detection of relative motion, i.e., motion contrast, has been reported for motion-sensitive neurons in several vertebrate systems, yet the mechanism underlying motion-contrast sensitivity remains unknown. An algorithm for computing motion contrast directly from the moving intensity distribution is proposed. In this algorithm, the time-dependent intensity distribution of the visual space is convolved with a periodic function. For coherent motion, the resulting convolution integral reduces to a traveling wave of fixed amplitude, while incoherent motion causes the amplitude to oscillate. The frequency of the amplitude oscillation provides a measure of motion contrast. The algorithm is successful in reproducing tuning curves derived from measurements of motion contrast sensitivity in avian tectum and primate middle temporal area. PMID- 15524550 TI - Epidemic spreading in a hierarchical social network. AB - A model of epidemic spreading in a population with a hierarchical structure of interpersonal interactions is described and investigated numerically. The structure of interpersonal connections is based on a scale-free network. Spatial localization of individuals belonging to different social groups, and the mobility of a contemporary community, as well as the effectiveness of different interpersonal interactions, are taken into account. Typical relations characterizing the spreading process, like a range of epidemic and epidemic curves, are discussed. The influence of preventive vaccinations on the spreading process is investigated. The critical value of preventively vaccinated individuals that is sufficient for the suppression of an epidemic is calculated. Our results are compared with solutions of the master equation for the spreading process and good agreement of the character of this process is found. PMID- 15524551 TI - Topological generalizations of network motifs. AB - Biological and technological networks contain patterns, termed network motifs, which occur far more often than in randomized networks. Network motifs were suggested to be elementary building blocks that carry out key functions in the network. It is of interest to understand how network motifs combine to form larger structures. To address this, we present a systematic approach to define "motif generalizations": families of motifs of different sizes that share a common architectural theme. To define motif generalizations, we first define "roles" in a subgraph according to structural equivalence. For example, the feedforward loop triad--a motif in transcription, neuronal, and some electronic networks--has three roles: an input node, an output node, and an internal node. The roles are used to define possible generalizations of the motif. The feedforward loop can have three simple generalizations, based on replicating each of the three roles and their connections. We present algorithms for efficiently detecting motif generalizations. We find that the transcription networks of bacteria and yeast display only one of the three generalizations, the multi output feedforward generalization. In contrast, the neuronal network of C. elegans mainly displays the multi-input generalization. Forward-logic electronic circuits display a multi-input, multi-output hybrid. Thus, networks which share a common motif can have very different generalizations of that motif. Using mathematical modeling, we describe the information processing functions of the different motif generalizations in transcription, neuronal, and electronic networks. PMID- 15524552 TI - Spectrum and symbol distribution of nucleotide sequences. AB - This paper explores the connection between the size of the spectral coefficients of a nucleotide or any other symbolic sequence and the distribution of nucleotides along certain subsequences. It explains the connection between the nucleotide distribution and the size of the spectral coefficients, and gives a necessary and sufficient condition for a coefficient to have a prescribed magnitude. Furthermore, it gives a fast algorithm for computing the value of a given spectral coefficient of a nucleotide sequence, discussing periods 3 and 4 as examples. Finally, it shows that the spectrum of a symbolic sequence is redundant, in the sense that there exists a linear recursion that determines the values of all the coefficients from those of a subset. PMID- 15524553 TI - Robust stochastic resonance for simple threshold neurons. AB - Simulation and theoretical results show that memoryless threshold neurons benefit from small amounts of almost all types of additive noise and so produce the stochastic-resonance or SR effect. Input-output mutual information measures the performance of such threshold systems that use subthreshold signals. The SR result holds for all possible noise probability density functions with finite variance. The only constraint is that the noise mean must fall outside a "forbidden" threshold-related interval that the user can control--a new theorem shows that this condition is also necessary. A corollary and simulations show that the SR effect occurs for right-sided beta and Weibull noise as well. These SR results further hold for the entire uncountably infinite class of alpha-stable probability density functions. Alpha-stable noise densities have infinite variance and infinite higher-order moments and often model impulsive noise environments. The stable noise densities include the special case of symmetric bell-curve densities with thick tails such as the Cauchy probability density. The SR result for alpha-stable noise densities shows that the SR effect in threshold and thresholdlike systems is robust against occasional or even frequent violent fluctuations in noise. Regression analysis reveals both an exponential relationship for the optimal noise dispersion as a function of the alpha bell curve tail thickness and an approximate linear relationship for the SR-maximal mutual information as a function of the alpha bell-curve tail thickness. PMID- 15524554 TI - Periodically varying externally imposed environmental effects on population dynamics. AB - Effects of externally imposed periodic changes in the environment on population dynamics are studied with the help of a simple model. The environmental changes are represented by the temporal and spatial dependence of the competition terms in a standard equation of evolution. Possible applications of the analysis are on the one hand to bacteria in Petri dishes and on the other to rodents in the context of the spread of the Hantavirus epidemic. The analysis shows that spatiotemporal structures emerge, with interesting features which depend on the interplay of separately controllable aspects of the externally imposed environmental changes. PMID- 15524555 TI - Multidelayed random walks: theory and application to the neolithic transition in Europe. AB - We present a model in which particles (or individuals of a biological population) disperse with a rest time between consecutive motions (or migrations) which may take several possible values from a discrete set. Particles (or individuals) may also react (or reproduce). We derive a new equation for the effective rest time T of the random walk. Application to the neolithic transition in Europe makes it possible to derive more realistic theoretical values for its wavefront speed than those following from the single-delayed framework presented previously [J. Fort and V. Mendez, Phys. Rev. Lett. 82, 867 (1999)]. The new results are consistent with the archaeological observations of this important historical process. PMID- 15524556 TI - Solitons in alpha-helical proteins. AB - We investigate some aspects of the soliton dynamics in an alpha-helical protein macromolecule within the steric Davydov-Scott model. Our main objective is to elucidate the important role of the helical symmetry in the formation, stability, and dynamical properties of Davydov's solitons in an alpha helix. We show, analytically and numerically, that the corresponding system of nonlinear equations admits several types of stationary soliton solutions and that solitons which preserve helical symmetry are dynamically unstable: once formed, they decay rapidly when they propagate. On the other hand, the soliton which spontaneously breaks the local translational and helical symmetries possesses the lowest energy and is a robust localized entity. We also demonstrate that this soliton is the result of a hybridization of the quasiparticle states from the two lowest degenerate bands and has an inner structure which can be described as a modulated multihump amplitude distribution of excitations on individual spines. The complex and composite structure of the soliton manifests itself distinctly when the soliton is moving and some interspine oscillations take place. Such a soliton structure and the interspine oscillations have previously been observed numerically [A. C. Scott, Phys. Rev. A 26, 578 (1982)]. Here we argue that the solitons studied by Scott are hybrid solitons and that the oscillations arise due to the helical symmetry of the system and result from the motion of the soliton along the alpha helix. The frequency of the interspine oscillations is shown to be proportional to the soliton velocity. PMID- 15524557 TI - Double-wavelet approach to study frequency and amplitude modulation in renal autoregulation. AB - Biological time series often display complex oscillations with several interacting rhythmic components. Renal autoregulation, for instance, involves at least two separate mechanisms both of which can produce oscillatory variations in the pressures and flows of the individual nephrons. Using double-wavelet analysis we propose a method to examine how the instantaneous frequency and amplitude of a fast mode is modulated by the presence of a slower mode. Our method is applied both to experimental data from normotensive and hypertensive rats showing different oscillatory patterns and to simulation results obtained from a physiologically based model of the nephron pressure and flow control. We reveal a nonlinear interaction between the two mechanisms that regulate the renal blood flow in the form of frequency and amplitude modulation of the myogenic oscillations. PMID- 15524558 TI - Spectra and waiting-time densities in firing resonant and nonresonant neurons. AB - The response of a neural cell to an external stimulus can follow one of two patterns: nonresonant neurons monotonically relax to the resting state after excitation while resonant ones show subthreshold oscillations. We investigate how these subthreshold properties of neurons affect their suprathreshold response. Conversely we ask the following: Can we distinguish between both types of neuronal dynamics using suprathreshold spike trains? The dynamics of neurons is given by stochastic FitzHugh-Nagumo and Morris-Lecar models either having a focus or a node as the stable fixed point. We determine numerically the spectral power density as well as the interspike interval density in response to random (noiselike) signals. We show that the information about the type of dynamics obtained from power spectra is of limited validity. In contrast, the interspike interval density provides a very sensitive instrument for the diagnostics of whether the dynamics has resonant or nonresonant properties. For the latter value, we formulate a fit formula and use it to reconstruct theoretically the spectral power density, which coincides with the numerically obtained spectra. We underline that the renewal theory is applicable to analysis of suprathreshold responses even of resonant neurons. PMID- 15524559 TI - Thermoelectricity in natural and synthetic hydrogels. AB - We describe a technique for measuring a Seebeck effect in gels and present data for three systems. Notably distinct signals are obtained for gel originating in the electrosensitive organs of marine sharks, synthetic collagen-based gel, and as a control, seawater, the gels' solvent. Only the gel of sharks shows a reversible thermoelectric signal. The difference between gel samples and seawater simply confirms that gels suppress mass transport. The difference between synthetic gel and the gel of sharks shows that the charged polymers of the shark gel restrict mass transport much more successfully than the polymers of the collagen gel, and we submit that this sort of ion localization is key to the emergence of thermoelectricity in a gelatinous substance. We compare the properties of the natural gel to those of established thermoelectrics. PMID- 15524560 TI - Sierpinski signal generates 1/f alpha spectra. AB - We investigate the row sum of the binary pattern generated by the Sierpinski automaton: Interpreted as a time series we calculate the power spectrum of this Sierpinski signal analytically and obtain a unique rugged fine structure with underlying power law decay with an exponent of approximately 1.15. Despite the simplicity of the model, it can serve as a model for 1/f(alpha) spectra in a certain class of experimental and natural systems such as catalytic reactions and mollusc patterns. PMID- 15524561 TI - Diffusion-controlled annihilation A+B-->0: the growth of an A-particle island from a localized A source in the B-particle sea. AB - We present the growth dynamics of an island of particles A injected from a localized A source into a sea of particles B and dying in the course of diffusion controlled annihilation A+B-->0. We show that in the one-dimensional (1D) case the island grows unlimitedly at any source strength Lambda, and the dynamics of its growth does not depend asymptotically on the diffusivity of B particles. In the 3D case the island grows only at Lambda> Lambda(c), achieving asymptotically a stationary state (static island). In the marginal 2D case the island unlimitedly grows at any Lambda but at Lambda< Lambda(*) the time of its formation becomes exponentially large. For all cases the numbers of surviving and dying A particles are calculated, and the scaling of the reaction zone is derived. PMID- 15524562 TI - Steady states of a microtubule assembly in a confined geometry. AB - We study the steady state of an assembly of microtubules in a confined volume, analogous to the situation inside a cell where the cell boundary forms a natural barrier to growth. We show that the dynamical equations for growing and shrinking microtubules predict the existence of two steady states, with either exponentially decaying or exponentially increasing distribution of microtubule lengths. We identify the regimes in parameter space corresponding to these steady states. In the latter case, the apparent catastrophe frequency near the boundary is found to be significantly larger than that in the interior. Both the exponential distribution of lengths and the increase in the catastrophe frequency near the cell margin is in excellent agreement with recent experimental observations. PMID- 15524563 TI - Stabilization of surface-immobilized enzymes using grafted polymers. AB - We introduce a two-dimensional lattice model of immobilization and stabilization of proteinlike polymers using grafted polymers. The protein is designed to have a specific bulk conformation reproducing a catalytic cleft of natural enzymes. Our model predicts a first order denaturing adsorption transition of free proteins. On the other hand, for an immobilized protein we observe a more gradual disappearance of the hydrophobic centers accompanied by adsorption. We show that, using hydrophilic grafted polymers of proper length and grafting density, the conformation as well as the hydrophobic centers of the protein can be restored. PMID- 15524564 TI - Complex dynamics of the formation of spatially localized standing structures in the vicinity of saddle-node bifurcations of waves in the reaction-diffusion model of blood clotting. AB - Local activation in a one-dimensional three-component reaction-diffusion model of blood clotting may lead to a formation of spatially localized standing structures (peaks) via several complex scenarios. In the first scenario, two concentration pulses first propagate from the site of activation, then stop and transform into peaks [Zarnitsina et al., Chaos 11, 57 (2001)]. Here, we examine this scenario, and also describe a different scenario of peak formation. In this scenario, two trigger waves propagate initially in opposite directions away from the site of activation. Then they stop and change direction of propagation toward each other to the activation site, where they interact and form a peak. Both of these scenarios of stable peak formation are observed in the vicinity of saddle-node bifurcation and may be viewed as a memory of the extinct wave modes. PMID- 15524565 TI - Attack vulnerability of scale-free networks due to cascading breakdown. AB - The possibility that a complex network can be brought down by attack on a single or a very few nodes through the process of cascading failures is of significant concern. Here we investigate a recent model for cascading failures in complex networks and uncover a phase-transition phenomenon in terms of the key parameter characterizing the node capacity. For parameter value below the phase-transition point, cascading failures can cause the network to disintegrate almost entirely. We obtain a theoretical estimate for the phase-transition point and provide numerical support. PMID- 15524566 TI - Universality of the optimal path in the strong disorder limit. AB - We study numerically the optimal paths in two and three dimensions on various disordered lattices in the limit of strong disorder. We find that the length l of the optimal path scales with geometric distance r , as l approximately r (d(opt) ) with d(opt) =1.22+/-0.01 for d=2 and 1.44+/-0.02 for d=3 , independent of whether the optimization is on a path of weighted bonds or sites, and independent of the lattice or its coordination number. Our finding suggests that the exponent d(opt) is universal, depending only on the dimension of the system. PMID- 15524567 TI - Percolation of unsatisfiability in finite dimensions. AB - The optimization of two-dimensional Boolean formulas is studied using percolation theory, rare region arguments, and boundary effects. In contrast with mean-field results, there is no satisfiability transition as the constraint density is varied, although there is a logical connectivity transition. In the disconnected phase, there is a transition in the solution time. The thermodynamic ground state for this NP-hard optimization problem is unique; local solutions can be adjoined to find the global ground state. These results have implications for the computational study of disordered materials. PMID- 15524568 TI - Where two fractals meet: the scaling of a self-avoiding walk on a percolation cluster. AB - The scaling properties of self-avoiding walks on a d -dimensional diluted lattice at the percolation threshold are analyzed by a field-theoretical renormalization group approach. To this end we reconsider the model of Phys. Rev. Lett. 63, 2819 (1989)] and argue that via renormalization its multifractal properties are directly accessible. While the former first order perturbation did not agree with the results of other methods our analytic result gives an accurate description of the available MC and exact enumeration data in a wide range of dimensions 2 -s(a) , s(b) --> s(b) for b not equala . This includes spin models in the presence of random cubic symmetric anisotropy with probability distribution vanishing outside the lattice axes. Using nonperturbative arguments we show the existence of a stable fixed point corresponding to the random-exchange Ising universality class. The field theoretical renormalization-group flow is investigated in the framework of a fixed-dimension expansion in powers of appropriate quartic couplings, computing the corresponding beta functions to five loops. This analysis shows that the random Ising fixed point is the only stable fixed point that is accessible from the relevant parameter region. Therefore, if the system undergoes a continuous transition, it belongs to the random-exchange Ising universality class. The approach to the asymptotic critical behavior is controlled by scaling corrections with exponent Delta= -alpha(r) , where alpha(r) approximately -0.05 is the specific-heat exponent of the random-exchange Ising model. PMID- 15524585 TI - Aging and rejuvenation with fractional derivatives. AB - We discuss a dynamic procedure that makes fractional derivatives emerge in the time asymptotic limit of non-Poisson processes. We find that two-state fluctuations, with an inverse power-law distribution of waiting times, finite first moment, and divergent second moment, namely, with the power index mu in the interval 2> t(a) yields o=mu-2 . The intermediate time regime is probably incompatible with a picture based on fractional derivatives, or, at least, with a mono-order fractional derivative. PMID- 15524586 TI - Self-organization of collaboration networks. AB - We study collaboration networks in terms of evolving, self-organizing bipartite graph models. We propose a model of a growing network, which combines preferential edge attachment with the bipartite structure, generic for collaboration networks. The model depends exclusively on basic properties of the network, such as the total number of collaborators and acts of collaboration, the mean size of collaborations, etc. The simplest model defined within this framework already allows us to describe many of the main topological characteristics (degree distribution, clustering coefficient, etc.) of one-mode projections of several real collaboration networks, without parameter fitting. We explain the observed dependence of the local clustering on degree and the degree degree correlations in terms of the "aging" of collaborators and their physical impossibility to participate in an unlimited number of collaborations. PMID- 15524587 TI - Minimizing energy below the glass thresholds. AB - Focusing on the optimization version of the random K -satisfiability problem, the MAX- K -SAT problem, we study the performance of the finite energy version of the survey propagation algorithm. We show that a simple (linear time) backtrack decimation strategy is sufficient to reach configurations well below the lower bound for the dynamic threshold energy and very close to the analytic prediction for the optimal ground states. A comparative numerical study on one of the most efficient local search procedures is also given. PMID- 15524588 TI - Networking the seceder model: Group formation in social and economic systems. AB - The seceder model illustrates how the desire to be different from the average can lead to formation of groups in a population. We turn the original, agent based, seceder model into a model of network evolution. We find that the structural characteristics of our model closely match empirical social networks. Statistics for the dynamics of group formation are also given. Extensions of the model to networks of companies are also discussed. PMID- 15524589 TI - Cluster growth in far-from-equilibrium particle models with diffusion, detachment, reattachment, and deposition. AB - Monolayer cluster growth in far-from-equilibrium systems is investigated by applying simulation and analytic techniques to minimal hard core particle (exclusion) models. The first model (I), for postdeposition coarsening dynamics, contains mechanisms of diffusion, attachment, and slow activated detachment (at rate epsilon<<1 ) of particles on a line. Simulation shows three successive regimes of cluster growth: fast attachment of isolated particles; detachment allowing further ( epsilont )(1/3) coarsening of average cluster size; and t( 1/2) approach to a saturation size varying as epsilon(-1/2) . Model II generalizes the first one in having an additional mechanism of particle deposition into cluster gaps, suppressed for the smallest gaps. This model exhibits early rapid filling, leading to slowing deposition due to the increasing scarcity of deposition sites, and then continued power law [ ( epsilont )(1/2) ] cluster size coarsening through the redistribution allowed by slow detachment. The basic ( epsilont )(1/3) domain growth laws and epsilon(-1/2) saturation in model I are explained by a simple scaling picture involving the time for a particle to detach and diffuse to the next cluster. A second, fuller approach is presented that employs a mapping of cluster configurations to a column picture and an approximate factorization of the cluster configuration probability within the resulting master equation. This allows, through the steady state solution of the corresponding equation for a cluster probability generating function, quantitative results for the saturation of model I in excellent agreement with the simulation results. For model II, it provides a one-variable scaling function solution for the coarsening probability distribution, and in particular quantitative agreement with the cluster length scaling and its amplitude. PMID- 15524590 TI - Statistical analysis of the extreme values of stress time series from the Portevin-Le Chatelier effect. AB - In an effort to understand the deterministic vs stochastic character of the Portevin-Le Chatelier (PLC) phenomenon, we investigate the structure of the underlying mechanism that generates the stick-slip patterns of stress over time. The stress time series is reduced to a series of successive pairs of minimum and maximum values representing the stick-slip patterns and a statistical analysis by means of hypothesis testing is applied to it. The null hypothesis of least deterministic structure is that the time series of extreme values is a bounded random walk of alternating direction (BRWAD); that is, besides the constraint of succession of minima to maxima bounded at a predefined range there are no other correlations in the data. To implement the test we use surrogate data generated by a model consistent with a BRWAD type process, which also uses the statistics of the original data to best mimic them. The proposed hypothesis testing is found to perform properly on simulated data from stochastic and deterministic systems. For the PLC time series, the null hypothesis is rejected at a high level of confidence giving evidence for some deterministic structure in the succession of the extreme stress values. This result allows for further statistical analysis including also the time aspect of the stick-slip patterns. PMID- 15524591 TI - Survival probability of a diffusing test particle in a system of coagulating and annihilating random walkers. AB - We calculate the survival probability of a diffusing test particle in an environment of diffusing particles that undergo coagulation at rate lambda(c) and annihilation at rate lambda(a) . The test particle is annihilated at rate lambda(') on coming into contact with the other particles. The survival probability decays algebraically with time as t(-theta;) . The exponent theta; in d<2 is calculated using the perturbative renormalization group formalism as an expansion in epsilon=2-d . It is shown to be universal, independent of lambda(') , and to depend only on delta , the ratio of the diffusion constant of test particles to that of the other particles, and on the ratio lambda(a) / lambda(c) . In two dimensions we calculate the logarithmic corrections to the power law decay of the survival probability. Surprisingly, the logarithmic corrections are nonuniversal. The one-loop answer for theta; in one dimension obtained by setting epsilon=1 is compared with existing exact solutions for special values of delta and lambda(a) / lambda(c) . The analytical results for the logarithmic corrections are verified by Monte Carlo simulations. PMID- 15524592 TI - Triangular Ising model with nearest- and next-nearest-neighbor couplings in a field. AB - The authors study the Ising model on the triangular lattice with nearest-neighbor couplings K(nn) , next-nearest-neighbor couplings K(nnn) >0 , and a magnetic field H . This work is done by means of finite-size scaling of numerical results of transfer matrix calculations, and Monte Carlo simulations. We determine the phase diagram and confirm the character of the critical manifolds. The emphasis of this work is on the antiferromagnetic case K(nn) <0 , but we also explore the ferromagnetic regime K(nn) >/=0 for H=0 . For K(nn) <0 and H=0 we locate a critical phase presumably covering the whole range -infinity< K(nn) <0 . For K(nn) <0 , H not equal 0 we locate a plane of phase transitions containing a line of tricritical three-state Potts transitions. In the limit H-->infinity this line leads to a tricritical model of hard hexagons with an attractive next-nearest neighbor potential. PMID- 15524593 TI - Stochastic process of equilibrium fluctuations of a system with long-range interactions. AB - The relaxation towards equilibrium of systems with long-range interactions is not yet understood. As a step towards such a comprehension, we propose the study of dynamical equilibrium fluctuations in a model system with long-range interaction. We compute analytically, from the microscopic dynamics, the autocorrelation function of the order parameter. From this result, we derive analytically a Fokker-Planck equation which describes the stochastic process of the impulsion of a single particle in an equilibrium bath. The diffusion coefficient is explicitly computed. PMID- 15524594 TI - Pair approximation of the stochastic susceptible-infected-recovered-susceptible epidemic model on the hypercubic lattice. AB - We investigate the time evolution and steady states of the stochastic susceptible infected-recovered-susceptible (SIRS) epidemic model on one- and two-dimensional lattices. We compare the behavior of this system, obtained from computer simulations, with those obtained from the mean-field approximation (MFA) and pair approximation (PA). The former (latter) approximates higher-order moments in terms of first- (second-) order ones. We find that the PA gives consistently better results than the MFA. In one dimension, the improvement is even qualitative. PMID- 15524595 TI - Anomalous scaling of superrough growing surfaces: from correlation functions to residual local interfacial widths and scaling exponents. AB - A study on the (1+1) -dimensional superrough growth processes is undertaken. We first work out the exact relations among the local interfacial width w , the correlation function G , and the pth degree residual local interfacial width w(p) with p=1,2,3,... . The relations obtained are exact and thus can be applied to any (1+1) -dimensional growth processes in the continuum limit, no matter whether the interface is superrough or not. Then we investigate the influence of the macroscopic structure formation on the scaling behavior of the superrough growth processes. Moreover, we show analytically that the residual local interfacial width w(p) excludes only the influence of the macroscopic structure on the scaling behavior of the system and retains the true scaling behavior originating from the stochastic nature of the system. Finally, we analyze and simulate some superrough growth models for demonstration. PMID- 15524596 TI - Phase transitions in moving systems. AB - The general stability criteria of the supercritical Ginzburg-Landau equations in moving media are considered for different forms of the convective velocity which may change in space and time both periodically and randomly. The results are correlated with experiments on the propagation of vortices in superconducting films under the influence of a bias current. The role of the finite size of a sample is discussed. PMID- 15524597 TI - Navigation in a small world with local information. AB - It is commonly known that there exist short paths between vertices in a network showing the small-world effect. Yet vertices, for example, the individuals living in society, usually are not able to find the shortest paths, due to the very serious limit of information. To study this issue theoretically, here the navigation process of launching messages toward designated targets is investigated on a variant of the one-dimensional small-world network (SWN). In the network structure considered, the probability of a shortcut falling between a pair of nodes is proportional to r(-alpha) , where r is the lattice distance between the nodes. When alpha=0 , it reduces to the SWN model with random shortcuts. The system shows the dynamic small-world effect, which is different from the well-studied static SW effect. We study the effective network diameter, the path length as a function of the lattice distance, and the dynamics. They are controlled by multiple parameters, and we use data collapse to show that the parameters are correlated. The central finding is that, in the one-dimensional network studied, the dynamic SW effect exists for 0 upsilon(0) , the ground state energy. PMID- 15524606 TI - Two-dimensional map for impact oscillator with drift. AB - An impact oscillator with drift is considered. The model accounts for viscoelastic impacts and is capable of mimicking the dynamics of progressive motion, which is important in many applications. To simplify the analysis of this system, a transformation decoupling the original coordinates is introduced. As a result, the bounded oscillations are separated from the drift motion. To study the bounded dynamics, a two-dimensional analytical map is developed and analyzed. In general, the dynamic state of the system is fully described by four variables: time tau , relative displacement p and velocity y of the mass, and relative displacement q of the slider top. However, this number can be reduced to two if the beginning of the progression phase is being monitored. The lower and upper bounds of the map domain are approximated. A graphical method of iteration of the two-dimensional map, similar to the cobweb method used in the one-dimensional case, is proposed. The results of numerical iterations of this two-dimensional map are presented, and a comparison is given between bifurcation diagrams calculated for this map and for the original system of differential equations. PMID- 15524607 TI - Rotating hexagonal pattern in a dielectric barrier discharge system. AB - Here, we report on the experimental observation of a rotating hexagonal pattern in a continuous dissipative medium. The system under investigation is a planar dielectric barrier gas-discharge cell. The pattern consists of a set of current filaments occupying the whole discharge area and rotating as a rigid body. The symmetry of the rotating hexagons is lower than the symmetry of the stationary hexagonal pattern. We study the dynamics of the pattern, especially peculiarities of its rotational velocity. The temperature of the gas is found to be an important quantity influencing the rotating hexagons. PMID- 15524608 TI - Stability of attractors formed by inertial particles in open chaotic flows. AB - Particles having finite mass and size advected in open chaotic flows can form attractors behind structures. Depending on the system parameters, the attractors can be chaotic or nonchaotic. But, how robust are these attractors? In particular, will small, random perturbations destroy the attractors? Here, we address this question by utilizing a prototype flow system: a cylinder in a two dimensional incompressible flow, behind which the von Karman vortex street forms. We find that attractors formed by inertial particles behind the cylinder are fragile in that they can be destroyed by small, additive noise. However, the resulting chaotic transient can be superpersistent in the sense that its lifetime obeys an exponential-like scaling law with the noise amplitude, where the exponent in the exponential dependence can be large for small noise. This happens regardless of the nature of the original attractor, chaotic or nonchaotic. We present numerical evidence and a theory to explain this phenomenon. Our finding makes direct experimental observation of superpersistent chaotic transients feasible and it also has implications for problems of current concern such as the transport and trapping of chemically or biologically active particles in large scale flows. PMID- 15524609 TI - Poincare Husimi representation of eigenstates in quantum billiards. AB - For the representation of eigenstates on a Poincare section at the boundary of a billiard different variants have been proposed. We compare these Poincare Husimi functions, discuss their properties, and based on this select one particularly suited definition. For the mean behavior of these Poincare Husimi functions an asymptotic expression is derived, including a uniform approximation. We establish the relation between the Poincare Husimi functions and the Husimi function in phase space from which a direct physical interpretation follows. Using this, a quantum ergodicity theorem for the Poincare Husimi functions in the case of ergodic systems is shown. PMID- 15524610 TI - Chaotic Hamiltonian ratchets for pulsed periodic double-well potentials: classical correlations and the ratchet current. AB - We present analytical derivations of the diffusion rates, ratchet currents, and time scales of a new ratchet in a fully chaotic Hamiltonian system, introduced in Phys. Rev. Lett. 89, 194102 (2002)] with a proposed implementation using atoms in pulsed standing waves of light. The origin of this type of ratchet current is in asymmetric momentum diffusion rates which result when a "double-well" lattice is pulsed with unequal "kick" periods. The form of the new short-time correlations which modify the diffusion rates are derived. The resulting formulas for the classical energy diffusion rates are shown to give good agreement with numerical simulations. A closed analytical formula for the ratchet current is also obtained, which predicts correctly the current magnitudes and current reversals. The characteristic "ratchet time," a classical time scale associated with the momentum-diffusion ratchet is derived analytically. The competition between the ratchet time t(r) , and the quantum break time t(*) is investigated further. PMID- 15524611 TI - Counting function for a sphere of anisotropic quartz. AB - We calculate the leading Weyl term of the counting function for a monocrystalline quartz sphere. In contrast to other studies of counting functions, the anisotropy of quartz is a crucial element in our investigation. Hence we do not obtain a simple analytical form, but we carry out a numerical evaluation. To this end we employ the Radon transform representation of the Green's function. We compare our result to a previously measured unique data set of several tens of thousands of resonances. PMID- 15524612 TI - Regularity and chaos in interacting two-body systems. AB - We study classical and quantum chaos for two interacting particles on the plane. This is the simplest nontrivial case which sheds light on chaos in interacting many-body systems. The system consists of a confining one-body potential, assumed to be a deformed harmonic oscillator, and a two-body interaction of Coulomb type. In general, the dynamics is mixed with regular and chaotic trajectories. The relative roles of the one-body field and the two-body interaction are investigated. Chaos sets in as the strength of the two-body interaction increases. However, the degree of chaoticity strongly depends on the shape of the one-body potential and, for some shapes of the harmonic oscillator, the dynamics remains regular for all values of the two-body interaction. Scaling properties are found for the classical as well as for the quantum mechanical problem. PMID- 15524613 TI - Coupling scheme for complete synchronization of periodically forced chaotic CO2 lasers. AB - We present a way of coupling two nonautonomous, periodically forced, chaotic C O2 lasers in a master-slave configuration in order to achieve complete synchronization. The method consists of modulating the forcing of the slave laser by means of the difference between the intensities of the two lasers, and lends itself to a simple physical implementation. Experimental evidence of complete synchronization induced by a suitable coupling strength is shown, and a numerical model is used to achieve further insight of the synchronization phenomena. Finally, we describe a possible application of the investigated technique to the design of a digital communication system. PMID- 15524614 TI - Lyapunov spectrum of the many-dimensional dilute random Lorentz gas. AB - For a better understanding of the chaotic behavior of systems of many moving particles, it is useful to look at other systems with many degrees of freedom. An interesting example is the high-dimensional Lorentz gas, which, just like a system of moving hard spheres, may be interpreted as a dynamical system consisting of a point particle in a high-dimensional phase space, moving among fixed scatterers. In this paper, we calculate the full spectrum of Lyapunov exponents for the dilute random Lorentz gas in an arbitrary number of dimensions. We find that the spectrum becomes flatter with increasing dimensionality. Furthermore, for fixed collision frequency the separation between the largest Lyapunov exponent and the second largest one increases logarithmically with dimensionality, whereas the separations between Lyapunov exponents of given indices not involving the largest one go to fixed limits. PMID- 15524615 TI - Domain dynamics in the anisotropic Swift-Hohenberg equation. AB - Two types of asymptotic ordering processes in the anisotropic Swift-Hohenberg equation are studied, paying particular attention to the interaction between domain walls. For the first type, we will discuss the time evolution in which the spatially oscillatory patterns are formed, and show that two kinds of patterns exist depending on whether or not the imaginary part of the field vanishes. When the imaginary part is present, the equation has two distinct states which are regarded as kinds of domains, so the dynamics between two domain walls is established. We then discuss, for the second type, the dynamics when nontrivial uniform states are constructed. There exist two different domain walls, the Ne el type wall and the Bloch type wall, in a similar way to the anisotropic Ginzburg Landau equation. The equation of motion for two domain walls is derived, and it is shown that the distance between the two domain walls eventually approaches a finite length. The theoretical result is confirmed by numerical simulations. This fact proves the validity of the prediction on the temporal development of the distance between two domain walls. PMID- 15524616 TI - Bifurcation study of regular pulse packages in laser diodes subject to optical feedback. AB - We study the influence of delayed optical feedback from a short external cavity on the emission dynamics of semiconductor lasers using the Lang and Kobayashi rate equation model. A combination of numerical integration and continuation techniques allows us to bring new light into the bifurcation scenario leading to the regular pulse packages (RPP) regime. We give examples of bistability between RPP and time-periodic or steady state solutions. Our bifurcation study of RPP regime is complemented by an analysis of the dependency of the RPP period on the laser and feedback parameters. We qualitatively study this new dynamical regime by plotting a two-dimensional map in the feedback parameters space. The occurrence of RPP is furthermore associated with a topological change in the bifurcation diagram and accompanied by the creation of a new type of bifurcation bridge between a mode and an antimode. PMID- 15524617 TI - Fractal rock slope dynamics anticipating a collapse. AB - Time series of dilatometric measurements of relative displacements on rock cracks on stable and unstable sandstone slopes were analyzed. The inherent dynamics of rock slopes lack any significant nonlinearity. However, the residuals obtained by removing meteorological influences are fat-tailed non-Gaussian fluctuations, with short-range correlations in the case of stable slopes. The fluctuations of unstable slopes exhibit self-affine dynamics of fractional Brownian motions with power-law long-range correlations and are characterized by asymptotic power-law probability distributions with decay coefficients outside the range of stable Levy distributions. PMID- 15524618 TI - Transient and stationary chaos of a Bose-Einstein condensate loaded into a moving optical lattice potential. AB - Chaotic space-time evolution is investigated for the particle number density of a Bose-Einstein condensate with attractive interatomic interaction loaded into a traveling optical lattice. Melnikov chaos is studied and the weakly chaotic regime is presented analytically. Transitions from transient to stationary chaos in the space-time evolution are illustrated numerically. The results show that, on increasing the strength of the optical potential, the transient chaos falls onto several different attractors. Meanwhile, these attractors undergo a series of period-doubling bifurcations when the optical potential intensity is increased continuously, and eventually stationary chaos arises for a critical depth of the optical lattice. The obstructions to chaos caused by the damping and the motion of lattice are also demonstrated. PMID- 15524619 TI - Classical-quantum correspondence for the scattering dwell time. AB - Using results from the theory of dynamical systems, we derive a general expression for the classical average scattering dwell time tau . Remarkably, tau depends only on a ratio of phase space volumes. We further show that, for a wide class of systems, the average classical dwell time is not in correspondence with the energy average of the quantum Wigner time delay. PMID- 15524620 TI - Hierarchical structure description of spatiotemporal chaos. AB - We develop a hierarchical structure (HS) analysis for quantitative description of statistical states of spatially extended systems. Examples discussed here include an experimental reaction-diffusion system with Belousov-Zhabotinsky kinetics, the two-dimensional complex Ginzburg-Landau equation, and the modified FitzHugh Nagumon equation, which all show complex dynamics of spirals and defects. We demonstrate that the spatial-temporal fluctuation fields in the above-mentioned systems all display the HS similarity property originally proposed for the study of fully developed turbulence [Phys. Rev. Lett. 72, 336 (1994)]]. The derived values of a HS parameter beta from experimental and numerical data in various physical regimes exhibit consistent trends and characterize the degree of turbulence in the systems near the transition, and the degree of heterogeneity of multiple disorders far from the transition. It is suggested that the HS analysis offers a useful quantitative description for the complex dynamics of two dimensional spatiotemporal patterns. PMID- 15524621 TI - Reactions in flows with nonhyperbolic dynamics. AB - We study the reaction dynamics of active particles that are advected passively by 2D incompressible open flows, whose motion is nonhyperbolic. This nonhyperbolicity is associated with the presence of persistent vortices near the wake, wherein fluid is trapped. We show that the fractal equilibrium distribution of the reactants is described by an effective dimension d(eff) , which is a finite resolution approximation to the fractal dimension. Furthermore, d(eff) depends on the resolution epsilon and on the reaction rate 1/tau . As tau is increased, the equilibrium distribution goes through a series of transitions where the effective dimension increases abruptly. These transitions are determined by the complex structure of Cantori surrounding the Kolmogorov-Arnold Moser (KAM) islands. PMID- 15524622 TI - Observation of robust quantum resonance peaks in an atom optics kicked rotor with amplitude noise. AB - The effect of pulse train noise on the quantum resonance peaks of the atom optics kicked rotor is investigated experimentally. Quantum resonance peaks in the late time mean energy of the atoms are found to be surprisingly robust against all levels of noise applied to the kicking amplitude, while even small levels of noise on the kicking period lead to their destruction. The robustness to amplitude noise of the resonance peak and of the fall-off in mean energy to either side of this peak are explained in terms of the occurrence of stable, epsilon classical dynamics [Nonlinearity 16, 1381 (2003)]] around each quantum resonance. PMID- 15524623 TI - Interaction of Ising-Bloch fronts with Dirichlet boundaries. AB - We study the Ising-Bloch bifurcation in two systems, the complex Ginzburg Landau equation (CGLE) and a FitzHugh Nagumo (FN) model in the presence of spatial inhomogeneity introduced by Dirichlet boundary conditions. It is seen that the interaction of fronts with boundaries is similar in both systems, establishing the generality of the Ising-Bloch bifurcation. We derive reduced dynamical equations for the FN model that explain front dynamics close to the boundary. We find that front dynamics in a highly nonadiabatic (slow front) limit is controlled by fixed points of the reduced dynamical equations, that occur close to the boundary. PMID- 15524624 TI - Synchronization of spatially extended chaotic systems in the presence of asymmetric coupling. AB - In a recent paper [Phys. Rev. Lett. 91, 064103 (2003)]] we described the effects of asymmetric coupling configurations on the synchronization of spatially extended systems. In this paper, we report the consequences induced by the presence of asymmetries in the coupling scheme on the synchronization process of a pair of one-dimensional fields obeying complex Ginzburg-Landau equations. While synchronization always occurs for large enough coupling strengths, asymmetries have the effect of enhancing synchronization and play a crucial role in setting the threshold for the appearance of the synchronized dynamics, as well as in selecting the statistical and dynamical properties of the synchronized motion. We analyze the process of synchronization in the presence of asymmetries when the dynamics is affected by the presence of phase singularities, and show that defects tend to anchor one system to the other. In addition, asymmetry controls the number of synchronized defects that are present in the dynamics. Possible consequences of such asymmetry induced effects in biological and natural systems are discussed. PMID- 15524625 TI - Effects of time-delayed feedback on chaotic oscillators. AB - We study the effects of time-delayed feedback on chaotic systems where the delay time is both fixed (static case) and varying (dynamic case) in time. For the static case, typical phase coherent and incoherent chaotic oscillators are investigated. Detailed phase diagrams are investigated in the parameter space of feedback gain ( K ) and delay time ( tau ). Linear stability analysis, by assuming the time-delayed perturbation, varies as e(lambdat) where lambda is the eigenvalue, gives the boundaries of the stability islands and critical feedback gains ( K(c) ) for both Rossler oscillators and Lorenz oscillators. We also found that the stability island are found when the delay time is about tau= (n+ 1 / 2 ) T , where n is an integer and T is the average period of the chaotic oscillator. It is shown that these analytical predictions agree well with the numerical results. For the dynamic case, we investigate Rossler oscillator with periodically modulated delay time. Stability regimes are found for parameter space of feedback gain and modulation frequency in which it was impossible to be stabilized for a fixed delay time. We also trace the detailed routes to the stability near the island boundaries for both cases by investigating bifurcation diagrams. PMID- 15524626 TI - Direction detector on an excitable field: field computation with coincidence detection. AB - Living organisms process information without any central control unit and without any ruling clock. We have been studying a novel computational strategy that uses a geometrically arranged excitable field, i.e., "field computation." As an extension of this research, in the present article we report the construction of a "direction detector" on an excitable field. Using a numerical simulation, we show that the direction of a input source signal can be detected by applying the characteristic as a "coincidence detector" embedded on an excitable field. In addition, we show that this direction detection actually works in an experiment using an excitable chemical system. These results are discussed in relation to the future development of "field computation." PMID- 15524627 TI - Conditional Lagrangian acceleration statistics in turbulent flows with Gaussian distributed velocities. AB - The random intensity of noise approach to the one-dimensional Laval-Dubrulle Nazarenko-type model having deductive support from the three-dimensional Navier Stokes equation is used to describe Lagrangian acceleration statistics of a fluid particle in developed turbulent flows. Intensity of additive noise and cross correlation between multiplicative and additive noises entering a nonlinear Langevin equation are assumed to depend on random velocity fluctuations in an exponential way. We use an exact analytic result for the acceleration probability density function obtained as a stationary solution of the associated Fokker Planck equation. We give a complete quantitative description of the available experimental data on conditional and unconditional acceleration statistics within the framework of a single model with a single set of fit parameters. The acceleration distribution and variance conditioned on Lagrangian velocity fluctuations and the marginal distribution calculated by using independent Gaussian velocity statistics are found to be in a good agreement with the recent high-Reynolds-number Lagrangian experimental data. The fitted conditional mean acceleration is very small, that is, in agreement with direct numerical simulations, and increases for higher velocities but it departs from the experimental data, which exhibit anisotropy of the studied flow. PMID- 15524628 TI - Reynolds stress model involving the mean spin tensor. AB - In this work, we develop a Reynolds stress model along the line of the approach presented by Commun. Nonlinear Sci. Numer. Simul. 9, 543 (2004)]], aiming to assess the role and contribution of the mean spin tensor in turbulence modeling. Here, the constitutive functional for the Reynolds stress depends on the mean spin tensor as well as the mean stretching tensor and its Jaumann derivative, the turbulent kinetic energy K , and the turbulent dissipation rate epsilon , which is at the complexity level of p=1,m=1 , and n=0 of a rate-type constitutive equation for the Reynolds stress proposed in the aforementioned paper. The explicit form for the Reynolds stress is obtained with recourse to the representation theorem and the theory of invariants developed in modern rational continuum mechanics, and, as an approximation, a nonlinear cubic K-epsilon model is worked out in which the model coefficients are analytically identified based on the experimental results of Tavoularis and Corrsin [J. Fluid Mech. 104, 311 (1981)]]. In addition, numerical results based on this model, in the forms of employing the Jaumann derivative and the Oldroyd derivative, respectively, for homogeneous turbulent shear flow and fully developed turbulent flow over a backward-facing step, are presented in comparison with those obtained based on a few previously proposed linear and nonlinear K-epsilon models, showing reasonably good agreement with the experimental results and the DNS data concerned and a better performance than the previously developed quadratic models. PMID- 15524629 TI - Nonlinear hydrodynamic waves: effects of the equation of state. AB - The problem of the decay of an initial discontinuity (the Riemann problem) is studied for a substance with abnormal properties when the rarefaction waves get a shock form, whereas the compression waves become nonsharp with the width proportional to the distance traveled. Such a situation is inherent to matter in a near-critical thermodynamic state and is also met in many other physical systems. The behavior of pressure jumps is compared for the van der Waals equation of state and for its more realistic three-parametric modification. It is shown that the evolution of the rarefaction and compression waves is strongly dependent on the value of the fundamental gasdynamic derivative determined by the equation of state. We demonstrate that for some substances with abnormal properties both rarefaction and compression waves can keep a shocklike form for a long period of time after discontinuity decay. PMID- 15524630 TI - Kelvin-Helmholtz instability for relativistic fluids. AB - We reexamine the stability of an interface separating two nonmagnetized relativistic fluids in relative motion, showing that, in an appropriate reference frame, it is possible to find analytic solutions to the dispersion relation. Moreover, we show that the critical value of the Mach number, introduced by compressibility, is unchanged from the nonrelativistic case if we redefine the Mach number as M= [beta/ (1- beta(2) )(1/2) ] [ beta(s) / (1- beta(2)(s) )(1/2) ](-1) , where beta and beta(s) are, respectively, the speed of the fluid and the speed of sound (in units of the speed of light). PMID- 15524631 TI - Electro-osmotic streaming on application of traveling-wave electric fields. AB - We describe ac electro-osmotic flow of an aqueous electrolyte on application of a traveling-wave electric field. Depending on the frequency of the applied traveling wave, the interaction of the electric double layer charge and the tangential electric field leads to fluid flow in the direction of the traveling wave. We have derived two theoretical models that describe this flow as a function of the amplitude of the applied electric potential, the signal frequency, and the material properties of the system. The first is based on a capacitative model and is limited to frequencies much lower than the double layer relaxation frequency. The second is an analytical solution of the electrokinetic equations and is also valid at higher frequencies. We provide experimental evidence that streaming takes place on application of a traveling wave of potential by tracing the movements of fluorescent latex beads over a spiral electrode structure. Streaming takes place at applied potentials low enough for the method to be easily integrated into lab-on-a-chip devices. PMID- 15524632 TI - Fractal aircraft trajectories and nonclassical turbulent exponents. AB - The dimension (D) of aircraft trajectories is fundamental in interpreting airborne data. To estimate D, we studied data from 18 trajectories of stratospheric aircraft flights 1600 km long taken during a "Mach cruise" (near constant Mach number) autopilot flight mode of the ER-2 research aircraft. Mach cruise implies correlated temperature and wind fluctuations so that DeltaZ approximately Deltax (H(z) ) where Z is the (fluctuating) vertical and x the horizontal coordinate of the aircraft. Over the range approximately 3-300 km , we found H(z) approximately 0.58+/-0.02 close to the theoretical 5/9=0.56 and implying D=1+ H(z) =14/9 , i.e., the trajectories are fractal. For distances <3 km aircraft inertia smooths the trajectories, for distances >300 km , D=1 again because of a rise of 1 m/km due to fuel consumption. In the fractal regime, the horizontal velocity and temperature exponents are close to the nonclassical value 1/2 (rather than 1/3 ). We discuss implications for aircraft measurements as well as for the structure of the atmosphere. PMID- 15524633 TI - 23/9 dimensional anisotropic scaling of passive admixtures using lidar data of aerosols. AB - In buoyancy-driven flows, another dimensional quantity appears in addition to the energy flux. Classically, this leads to the prediction that at large scales, isotropic Bolgiano-Obukhov (BO) scaling can dominate isotropic Kolmogorov scaling. We investigate this in the atmosphere by using state-of-the-art high powered lidar data. We examine simultaneous horizontal and vertical sections of passive scalar surrogates over the ranges 100 m to 120 km and 3 m to 4.5 km , respectively. Overall, this spans the crucial "mesoscale" and involves nearly 1000 times more data than the largest relevant experiments to date. Rather than a transition from one isotropic regime to another, we find that the two regimes always coexist in an anisotropic Corrsin-Obukhov law with the Kolmogorov holding in the horizontal, and the BO holding in the vertical. The stratification is quantified by an elliptical dimension D(el) found to be equal to 2.55+/-0.02 . This anisotropic scaling is very close to that predicted by the 23/9 dimensional unified scaling model of the atmosphere and is consistent with observations of the horizontal wind. PMID- 15524634 TI - Molecular to fluid dynamics: the consequences of stochastic molecular motion. AB - The derivation of fluid dynamic equations from molecular equations is considered. This is done on the basis of a stochastic model for the molecular motion which can be obtained by a projection of underlying deterministic equations. The stochastic model is used to derive fluid dynamic equations where the molecular stress tensor and heat flux appear as unknowns. However, the stochastic model also implies transport equations for these quantities. Combined with the assumption of a local equilibrium state, these transport equations can be used to derive a hierarchy of algebraic expressions for the molecular stress tensor and heat flux. A scaling analysis then explains the range of applicability of the Navier-Stokes model. The latter is relevant, for example, to simulations of high Mach-number turbulent flows. PMID- 15524635 TI - Two-point closure strategy in the mapping closure approximation approach. AB - A two-point closure strategy in mapping closure approximation (MCA) approach is developed for the evolution of the probability density function (PDF) of a scalar advected by stochastic velocity fields. The MCA approach is based on multipoint statistics. We formulate a MCA modeled system using the one-point PDFs and two point correlations. The MCA models can describe both the evolution of the PDF shape and the rate at which the PDF evolves. PMID- 15524636 TI - Polymers suppress the inverse transfers of energy and the enstrophy flux fluctuations in two-dimensional turbulence. AB - The addition of minute amounts of a flexible polymer to two-dimensional turbulence produced in fast-flowing soap films affects large scales and small scales differently. For large scales, the inverse transfers of energy are suppressed. For small scales, where mean quantities are barely affected, the enstrophy flux fluctuations are significantly reduced, making the flow less chaotic. PMID- 15524637 TI - Adhesion phenomena in ferrofluids. AB - One efficient way of determining the bond strength of adhesives is to measure the force or the work required to separate two surfaces bonded by a thin adhesive film. We consider the case in which the thin film is not a conventional adhesive material but a high viscosity ferrofluid confined between two narrowly spaced parallel flat plates subjected to an external magnetic field. Our theoretical results demonstrate that both the peak adhesive force and the separation energy are significantly influenced by the action and symmetry properties of the applied field. Specifically, we show that the adhesive strength of a ferrofluid is reduced if the applied magnetic field is perpendicular to the plates or if the applied field is in plane and exhibits azimuthal symmetry. Conversely, the adhesive strength can be either enhanced or reduced if the applied field is in plane and is directed radially outward. This establishes an interesting connection between adhesion and ferrohydrodynamic phenomena, allowing the control of important adhesive properties by magnetic means. PMID- 15524638 TI - Frozen water waves over rough topographical bottoms. AB - The propagation of surface water waves over rough topographical bottoms is investigated by the multiple scattering theory. It is shown that the waves can be localized spatially through the process of multiple scattering and wave interference, a peculiar wave phenomenon which has been previously discussed for frozen light in optical systems [Nature 390, 661 (1997)]]. This paper demonstrates that when frozen, the transmission of the waves falls off exponentially, and a cooperative behavior appears, fully supporting previous predictions. A graphic method is used to illustrate this distinct phase state in the wave propagation. PMID- 15524639 TI - Wave-number selection by target patterns and sidewalls in Rayleigh-Benard convection. AB - We present experimental results for patterns of Rayleigh-Benard convection in a cylindrical container with static sidewall forcing. The fluid used was methanol, with a Prandlt number sigma=7.17 , and the aspect ratio was Gamma identical withR/d approximately 19 ( R is the radius and d the thickness of the fluid layer). In the presence of a small heat input along the sidewall, a sudden jump of the temperature difference DeltaT from below to slightly above a critical value Delta T(c) produced a stable pattern of concentric rolls (a target pattern) with the central roll (the umbilicus) at the center of the cell. A quasistatic increase of epsilon identical withDeltaT/Delta T(c) -1 beyond epsilon(1,c) approximately 0.8 caused the umbilicus of the pattern to move off center. As observed by others, a further quasistatic increase of epsilon up to epsilon=15.6 caused a sequence of transitions at epsilon(i,b) ,i=1,...,8 , each associated with the loss of one convection roll at the umbilicus. Each loss of a roll was preceded by the displacement of the umbilicus away from the center of the cell. After each transition the umbilicus moved back toward but never quite reached the center. With decreasing epsilon new rolls formed at the umbilicus when epsilon was reduced below epsilon(i,a) < epsilon(i,b) . When decreasing epsilon , large umbilicus displacements did not occur. In addition to quantitative measurements of the umbilicus displacement, we determined and analyzed the entire wave director field of each image. The wave numbers varied in the axial direction, with minima at the umbilicus and at the cell wall and a maximum at a radial position close to 2Gamma/3 . The wave numbers at the maximum showed hysteretic jumps at epsilon(i,b) and epsilon(i,a) , but on average agreed well with the theoretical predictions for the wave numbers selected in the far field of an infinitely extended target pattern. To our knowledge there is as yet no prediction for the wave number selected by the umbilicus itself, or by the cell wall of the finite experimental system. PMID- 15524640 TI - Electron acceleration to energies beyond GeV by a relativistic ion beam instability. AB - Synchrotron emission suggests the presence of TeV electrons at various astrophysical objects. We propose a mechanism for the acceleration of electrons to ultrahigh energies (UHE) by intense electrostatic waves (ESWs). The latter are driven by dense proton beams that move at relativistic speeds relative to a background plasma and the electrons are accelerated by their nonlinear interaction with the ESWs. We follow the evolution of the wave instability by means of particle-in-cell (PIC) simulations. After the instability has saturated, we obtain spatially confined electron voids in which secondary instabilities develop due to resonant interactions between the beams and the background protons, generating intense ESWs which accelerate electrons to ultrarelativistic speeds within times of a few hundred inverse plasma frequencies. PMID- 15524641 TI - Effects of the electron energy distribution function on modeled x-ray spectra. AB - This paper presents the results of a broad investigation into the effects of the electron energy distribution function on the predictions of nonlocal thermodynamic equilibrium collisional-radiative atomic kinetics models. The effects of non-Maxwellian and suprathermal ("hot") electron distributions on collisional rates (including three-body recombination) are studied. It is shown that most collisional rates are fairly insensitive to the functional form and the characteristic (central or average) energy of the electron distribution function as long as the characteristic energy is larger than the threshold energy for the collisional process. Collisional excitation and ionization rates are, however, highly sensitive to the number of hot electrons. This permits the development of robust spectroscopic diagnostics that can be used to characterize the electron density, bulk electron temperature, and hot electron fraction of plasmas with nonequilibrium electron distribution functions. Hot electrons are shown to increase and spread out plasma charge state distributions, amplify the intensities of emission lines fed by direct collisional excitation and radiative cascades, and alter the structure of satellite features in both K - and L -shell spectra. The characteristic energy, functional form, and spatial properties of hot electron distributions in plasmas are open to characterization through their effects on high-energy continuum and line emission and on the polarization of spectral lines. PMID- 15524642 TI - Ultrashort relativistic electromagnetic solitons. AB - Ultrashort high-intensity electromagnetic solitons in both underdense and overdense plasmas are investigated. Comparison is made for solitons with smooth and sharp electron density profiles. It is found that subcycle relativistic solitons can propagate from low-density to high-density plasmas. PMID- 15524643 TI - Closed fluid description of relativistic, magnetized plasma interacting with radiation field. AB - A closed set of averaged fluid equations for a relativistic plasma immersed, simultaneously, in a slowly varying magnetizing field and a sharply varying electromagnetic field (radiation field, for example) of arbitrary intensity is derived. The modifications due to the radiation field on the plasma stress tensor and the Lorentz force are explicitly displayed. The resulting equations include the effects of radiation reaction as well as radiation pressure. PMID- 15524644 TI - Characterization of proton and heavier ion acceleration in ultrahigh-intensity laser interactions with heated target foils. AB - Proton and heavy ion acceleration in ultrahigh intensity ( approximately 2 x 10(20) W cm(-2) ) laser plasma interactions has been investigated using the new petawatt arm of the VULCAN laser. Nuclear activation techniques have been applied to make the first spatially integrated measurements of both proton and heavy ion acceleration from the same laser shots with heated and unheated Fe foil targets. Fe ions with energies greater than 10 MeV per nucleon have been observed. Effects of target heating on the accelerated ion energy spectra and the laser-to-ion energy conversion efficiencies are discussed. The laser-driven production of the long-lived isotope (57 )Co (271 days) via a heavy ion induced reaction is demonstrated. PMID- 15524645 TI - Normal modes of a quasi-one-dimensional multichain complex plasma. AB - We studied equally charged particles, suspended in a complex plasma, which move in a plane and interact with a screened Coulomb potential (Yukawa type) and with an additional external confining parabolic potential in one direction, which makes the system quasi-one-dimensional (Q1D). The normal modes of the system are studied in the presence of dissipation. We also investigated how a perpendicular magnetic field couples the phonon modes with each other. Two different ways of exciting the normal modes are discussed: (1) a uniform excitation of the Q1D lattice, and (2) a local forced excitation of the system in which one particle is driven by, e.g., a laser. Our results are in very good agreement with recent experimental findings on a finite single chain system [Phys. Rev. Lett. 91, 255003 (2003)]]. Predictions are made for the normal modes of multichain structures in the presence of damping. PMID- 15524646 TI - Large-amplitude oscillations and chaos in a Hamiltonian plasma system with many degrees of freedom. AB - Taking the beam-plasma system as a reference Hamiltonian system with many degrees of freedom, the connection between the development of large amplitude oscillations and stochasticity of the system (measured through the time-dependent maximum Lyapunov exponent) is investigated. It is found that the development of self-consistent large amplitude oscillations occurs in correspondence with the onset of chaos, and is related to a well defined change of topology of the phase space of the system. It is also shown that in a (Hamiltonian) linearly stable regime the development of large amplitude oscillations can occur when weakly dissipative processes are introduced. PMID- 15524647 TI - Suppression of small scale dynamo action by an imposed magnetic field. AB - Nonhelical hydromagnetic turbulence with an externally imposed magnetic field is investigated using direct numerical simulations. It is shown that the imposed magnetic field lowers the spectral magnetic energy in the inertial range. This is explained by a suppression of the small scale dynamo. At large scales, however, the spectral magnetic energy increases with increasing imposed field strength for moderately strong fields, and decreases only slightly for even stronger fields. The presence of Alfve n waves is explicitly confirmed by monitoring the evolution of magnetic field and velocity at one point. The frequency omega agrees with v(A) k(1) , where v(A) is the Alfve n speed and k(1) is the smallest wave number in the box. PMID- 15524648 TI - Kinetics of ultrashort relativistic electron pulses emitted from solid targets. AB - Interaction of ultrashort high-intensity laser pulses with solid targets generates relativistic electrons which escape from the target. The kinetics of these ultrashort electron pulses is governed by self-fields generated by the charge of the electron cloud. In this paper an analytical theory is developed which allows calculation of electron trajectories, electron fluxes, and electron spectra at any distance from the target. The theory is exact for two limiting cases: (a) a monoenergetic electron pulse with an arbitrary temporal shape; (b) an infinitely short electron pulse with an arbitrary energy spectrum. These results have applications in high-intensity irradiation experiments, e.g., in experiments irradiating samples with ultrashort electron or x-ray pulses, in developing optics for fourth-generation light sources, and in work relating to x ray lasers. PMID- 15524649 TI - Mechanism of electron multipacting with a long-bunch proton beam. AB - The energy gain and motion of electrons can quantitatively describe the mechanism of electron multipacting in a long-bunched proton machine. Strong multipacting usually happens around the bunches' tails due to the high energy of electrons when they hit the chamber surface. We investigated several important parameters of electron multipacting, proving that it is sensitive to the beam's intensity, the shape of its longitudinal profile, its transverse size, the secondary emission yield, and the energy at peak secondary emission yield. Our analyses, simulations, and experiments are all in agreement. PMID- 15524650 TI - Anisotropic enhanced backscattering induced by anisotropic diffusion. AB - The enhanced backscattering cone displaying a strong anisotropy from a material with anisotropic diffusion is reported. The constructive interference of the wave is preserved in the helicity preserving polarization channel and completely lost in the nonpreserving one. The internal reflectivity at the interface modifies the width of the backscatter cone. The reflectivity coefficient is measured by angular-resolved transmission. This interface property is found to be isotropic, simplifying the backscatter cone analysis. The material used is a macroporous semiconductor, gallium phosphide, in which pores are etched in a disordered position but with a preferential direction. PMID- 15524651 TI - Saturation-induced coherence loss in coherent backscattering of light. AB - We use coherent backscattering of light by cold strontium atoms to study phase breaking mechanisms in the multiple-scattering regime. As the probe light intensity is increased, the atomic optical transition starts to be saturated. Nonlinearities and inelastic scattering then occur. The latter induces a characteristic phase-breaking time that reduces the wave coherence. In our experiment, this leads to a strong reduction of the enhancement factor of the coherent backscattering cone. The results at different probe detuning are also presented. PMID- 15524652 TI - Circular photonic crystal resonators. AB - We study analytically and numerically a new class of circular resonators based on a radial photonic crystal reflector. The Bragg confinement enables the realization of compact resonators exhibiting both large free spectral range and high Q -factor. The dependence of the resonator characteristics on the reflector architecture and dimensions is studied in detail. Good agreement is found between the analytical and the numerical results obtained by finite-difference time domain simulations. PMID- 15524653 TI - Nonlinear optical pulse propagation simulation: from Maxwell's to unidirectional equations. AB - Spatial- and time-domain versions of the unidirectional pulse propagation equation (UPPE) are derived and compared from the point of view of their practical application in simulations of nonlinear optical pulse dynamics. A modification of the UPPE suitable for ultrathin optical waveguides, such as submicron silica wires, is also presented. We show in detail how various, previously published propagation equations follow from the UPPE in a unified way that clearly elucidates their underlying approximations and areas of applicability. PMID- 15524654 TI - Optical properties of an ideal homogeneous causal left-handed material slab. AB - The optical properties of a homogeneous slab of material characterized by causal permittivity epsilon (f) , and permeability mu (f) are investigated through finite difference time domain simulations. Lorentzian epsilon (f) and mu (f) are used to produce values of interest in the resulting index n (f) , namely, n<0 , 0infinity limit, we show that lambda(')(c)=d+2 and phi=z/2. We give a heuristic argument suggesting that this result is, in fact, valid for any dimension d and spin vector dimension n. We present numerical simulations for the conserved Ising model in d=1 and d=2, which are fully consistent with the present theory. PMID- 15524697 TI - Density functional theory for general hard-core lattice gases. AB - We put forward a general procedure to obtain an approximate free-energy density functional for any hard-core lattice gas, regardless of the shape of the particles, the underlying lattice, or the dimension of the system. The procedure is conceptually very simple and recovers effortlessly previous results for some particular systems. Also, the obtained density functionals belong to the class of fundamental measure functionals and, therefore, are always consistent through dimensional reduction. We discuss possible extensions of this method to account for attractive lattice models. PMID- 15524698 TI - Yang-Lee zeros of the antiferromagnetic Ising model. AB - There exists the famous circle theorem on the Yang-Lee zeros of the ferromagnetic Ising model. However, the Yang-Lee zeros of the antiferromagnetic Ising model are much less well understood than those of the ferromagnetic model. The precise distribution of the Yang-Lee zeros of the antiferromagnetic Ising model only with nearest-neighbor interaction J on LxL square lattices is determined as a function of temperature a=e(2betaJ) (J<0), and its relation to the phase transitions is investigated. In the thermodynamic limit (L-->infinity), the distribution of the Yang-Lee zeros of the antiferromagnetic Ising model cuts the positive real axis in the complex x=e(-2betaH) plane, resulting in the critical magnetic field +/ H(c)(a), where H(c)>0 below the critical temperature a(c)=square root of 2-1. The results suggest that the value of the scaling exponent y(h) is 1 along the critical line for api0mu+nu decays. AB - A search for T-violating transverse muon polarization (P(T)) in the K+- >pi(0)mu(+)nu decay was performed using kaon decays at rest. A new improved value P(T)=-0.0017+/-0.0023(stat)+/-0.0011(syst) was obtained giving an upper limit |P(T)|<0.0050. The T-violation parameter was determined to be Imxi=-0.0053+/ 0.0071(stat)+/-0.0036(syst) giving an upper limit |Imxi|<0.016. PMID- 15524702 TI - All conjugate-maximal-helicity-violating amplitudes from topological open string theory in twistor space. AB - It has recently been proposed that the D-instanton expansion of the open topological B model on P(3|4) is equivalent to the perturbative expansion of the maximally supersymmetric Yang-Mills theory in four dimensions. In this letter we show how to construct the gauge theory results for all n-point conjugate-maximal helicity-violating amplitudes by computing the integral over the moduli space of curves of degree n-3 in P(3|4), providing strong support to the string theory construction. PMID- 15524704 TI - Evidence for B0-->rho0pi0. AB - We present the first evidence of the decay B0-->rho(0)pi(0), using 140 fb(-1) of data collected at the Upsilon(4S) resonance with the Belle detector at the KEKB asymmetric e(+)e(-) collider. We detect 15.1+/-4.8 signal events with a significance of 3.5 standard deviations and measure the branching fraction to be B(B0-->rho(0)pi(0))=(5.1+/-1.6(stat)+/-0.9(syst))x10(-6). PMID- 15524705 TI - Evidence for B+-->omegal+nu. AB - We have searched for the decay B+-->omegal(+)nu (l=e or mu) in 78 fb(-1) of Upsilon(4S) data (85x10(6)BB events) accumulated with the Belle detector. The final state is fully reconstructed using the omega decay into pi(+)pi(-)pi(0), combined with detector hermeticity to estimate the neutrino momentum. A signal of 414+/-125 events is found in the data, corresponding to a branching fraction of (1.3+/-0.4+/-0.2+/-0.3)x10(-4), where the first two errors are statistical and systematic, respectively. The third error reflects the estimated form-factor uncertainty. PMID- 15524708 TI - Melting pattern of diquark condensates in quark matter. AB - Thermal color superconducting phase transitions in high density three-flavor quark matter are investigated in the Ginzburg-Landau approach. The effects of nonzero strange quark mass, electric and color charge neutrality, and direct instantons are considered. Weak coupling calculations show that an interplay between the mass and electric neutrality effects near the critical temperature gives rise to three successive second-order phase transitions as the temperature increases: a modified color-flavor locked (mCFL) phase (ud, ds, and us pairings) -> a d-quark superconducting (dSC) phase (ud and ds pairings) --> an isoscalar pairing phase (ud pairing) --> a normal phase (no pairing). The dSC phase is novel in the sense that while all eight gluons are Meissner screened as in the mCFL phase, three out of nine quark quasiparticles are always gapless. PMID- 15524709 TI - QCD vacuum as a disordered medium: a simplified model for the QCD dirac operator. AB - We model the QCD Dirac operator as a power-law random banded matrix (RBM) with the appropriate chiral symmetry. Our motivation is the form of the Dirac operator in a basis of instantonic zero modes with a corresponding gauge background of instantons. We compare the spectral correlations of this model to those of an instanton liquid model (ILM) and find agreement well beyond the Thouless energy. In the bulk of the spectrum the dimensionless Thouless energy of the RBM scales with the square root of system size in agreement with the ILM and chiral perturbation theory. Near the origin the scaling in the RBM remains the same as in the bulk which agrees with chiral perturbation theory but not with the ILM. Finally we discuss how this RBM should be modified in order to describe the spectral correlations of the QCD Dirac operator at the finite temperature chiral restoration transition. PMID- 15524711 TI - Experimental confirmation of the Alhassid-Whelan arc of regularity. AB - Evidence is presented to show that a group of nuclei, spanning a range of structures, corresponds to a previously proposed isolated region of regular behavior between vibrational and rotational structures that was never before observed empirically. Nuclei predicted to show such regular spectra correspond to Hamiltonian parameters that lie amidst those giving more chaotic spectra. We identify a key observable that has a one-to-one correspondence to this arc of regularity and which therefore provides both an empirical signature for it and a clue to its underlying nature. PMID- 15524712 TI - Quark structure and nuclear effective forces. AB - We formulate the quark meson coupling model as a many-body effective Hamiltonian. This leads naturally to the appearance of many-body forces. We investigate the zero range limit of the model and compare its Hartree-Fock Hamiltonian to that corresponding to the Skyrme effective force. By fixing the three parameters of the model to reproduce the binding and symmetry energy of nuclear matter, we find that it allows a very satisfactory interpretation of the Skyrme force. PMID- 15524710 TI - Measurement of the exclusive 3He(e,e'p) reaction below the quasielastic peak. AB - New, high-precision measurements of the 3He(e,e(')p) reaction using the A1 Collaboration spectrometers at the Mainz microtron MAMI are presented. These were performed in antiparallel kinematics at energy transfers below the quasielastic peak, and at a central momentum transfer of 685 MeV/c. Cross sections and distorted momentum distributions were extracted and compared to theoretical predictions and existing data. The longitudinal and transverse behavior of the cross section was also studied. Sizable differences in the cross-section behavior from theoretical predictions based on the plane wave impulse approximation were observed in both the two- and three-body breakup channels. Full Faddeev-type calculations account for some of the observed excess cross-section, but significant differences remain. PMID- 15524713 TI - Distribution of spectral widths and preponderance of spin-0 ground states in nuclei. AB - We use a single j-shell model with random two-body interactions to derive closed expressions for the distribution of and the correlations between spectral widths of different spins. This task is facilitated by introducing two-body operators whose squared spectral widths sum up to the squared spectral width of the random Hamiltonian. The spin-0 width is characterized by a relatively large average value and small fluctuations, while the width of maximum spin has the largest average and the largest fluctuations. The approximate proportionality between widths and spectral radii explains the preponderance of spin-0 ground states. PMID- 15524714 TI - Breakup densities of hot nuclei. AB - Breakup densities of hot 197Au-like residues have been deduced from the systematic trends of Coulomb parameters required to fit intermediate-mass fragment kinetic-energy spectra. The results indicate emission from nuclei near normal nuclear density below an excitation energy E(*)/A less, similar 2 MeV, followed by a gradual decrease to a near-constant value of rho/rho(0) approximately 0.3 for E(*)/A greater, similar 5 MeV. Temperatures derived from these data with a density-dependent Fermi-gas model yield a nuclear caloric curve that is generally consistent with those derived from isotope ratios. PMID- 15524715 TI - Caloric curve for mononuclear configurations. AB - The caloric curve for mononuclear configurations is studied with a schematic model. We investigate the dependence of the entropy on the density and effective mass profiles. In finite nuclei, a plateau in the caloric curve is a consequence of decreasing density and the destruction of correlations rather than an indication of phase coexistence. The mononuclear regime is metastable with respect to binary fission at low excitation energy and with respect to multifragmentation at high excitation. The statistical framework presented here is suitable to treat scenarios where experimental conditions are set to favor a population of highly excited mononuclei. PMID- 15524716 TI - Measurement of the electric quadrupole moment of the 4d2D5/2 level in 88Sr+. AB - The quadrupole moment of the 4d (2)D(5/2) level in 88Sr+ has been measured to be 2.6(3)ea(2)(0), where a(0) is the Bohr radius and e the elementary charge. A single laser-cooled strontium ion was confined in an end cap trap with a variable dc quadrupole potential, and measurements were made on the 5s (2)S(1/2)-4d (2)D(5/2) transition at 674 nm using a femtosecond optical frequency comb. This work shows that measurements of the unperturbed 88Sr+ transition frequency with sub-Hz uncertainty are possible and is important in understanding the reproducibility of ion trap optical frequency standards. PMID- 15524717 TI - Alternative low-symmetry structure for 13-atom metal clusters. AB - The atomic geometry, electronic structure, and magnetic moment of 4d transition metal clusters with 13 atoms are studied by pseudopotential density-functional calculations. We find a new buckled biplanar structure with a C(2v) symmetry stabilized by enhanced s-d hybridization. It has a lower energy than the close packed icosahedral or cuboctahedral structure for elements with more than half filled d shells. The magnetic moments of this buckled biplanar structure are found to be smaller than those of the icosahedral structure and closer to available experimental results. PMID- 15524718 TI - Collective polarization exchanges in collisions of photon clouds. AB - The one-loop "vacuum" Heisenberg-Euler coupling of four electromagnetic fields can lead to interesting collective effects in the collision of two photon clouds on a time scale order of magnitude faster than one estimates from the cross section and density. We estimate the characteristic time for macroscopic transformation of positive to negative helicity in clouds that are initially totally polarized and for depolarization of a polarized beam traversing an unpolarized cloud. PMID- 15524719 TI - Nonlinear frequency conversion in waveguide directional couplers. AB - Nonlinear optical effects for frequency conversion require a phase-matching condition to efficiently generate a coherent field at the new wavelength. We find that the phase-matching condition can be replaced by a resonance condition when the nonlinear effect takes place in a waveguide directional coupler. We apply this theory to second-harmonic generation and find a theoretical conversion efficiency of 100%, equivalent to the perfect phase-matching condition. An example of the design of such waveguide directional coupler is presented. PMID- 15524706 TI - Search for B+/--->[K(-/+)pi(+/-)](D)K+/- and upper limit on the b-->u amplitude in B+/--->DK+/-. AB - We search for B+/--->[K(-/+)pi(+/-)](D)K+/- decays, where [K(-/+)pi(+/-)](D) indicates that the K-/+pi(+/-) pair originates from the decay of a D0 or D (0). Results are based on 120x10(6) Upsilon(4S)-->BB decays collected with the BABAR detector at SLAC. We set an upper limit on the ratio R(Kpi) identical with[Gamma(B+-->[K(-)pi(+)](D)K+)+Gamma(B--->[K(+)pi(-)](D)K-)][Gamma(B+- >[K(+)pi(-)](D) / K+)+Gamma(B--->[K(-)pi(+)](D)K-)]<0.026 (90% C.L.). This constrains the amplitude ratio r(B) identical with|A(B--->D 0K-)/A(B--->D0K )|<0.22 (90% C.L.), consistent with expectations. The small value of r(B) favored by our analysis suggests that the determination of the Cabibbo-Kobayashi-Maskawa phase gamma from B-->DK will be difficult. PMID- 15524721 TI - Phase-coupling-induced ultraslow light propagation in solids at room temperature. AB - We show that the group velocities of light pulses can be decelerated dramatically by the use of a dispersive phase-coupling effect through a wave mixing process. We have observed experimentally such a phase-coupling-induced ultraslow light propagation with a group velocity as low as 0.05 m/s in a photorefractive Bi12SiO20 crystal at room temperature. Moreover, the ultraslow light is amplified in the Bi12SiO20 crystal because of the unidirectional energy transfer from a coupling beam to the ultraslow light. This technique to produce ultraslow light propagation is valid for all nonlinear wave mixing processes with a dispersive phase-coupling effect. PMID- 15524703 TI - Direct CP violating asymmetry in B0-->K+pi- decays. AB - We present a measurement of the direct CP violating asymmetry in the decay B0- >K+pi(-) using a data sample of 227x10(6) Upsilon(4S)-->BB decays collected with the BABAR detector at the PEP-II asymmetric-energy e(+)e(-) collider at SLAC. We observe a total signal yield of nK-(pi(+))+nK+(pi(-))=1606+/-51 decays and measure the asymmetry (nK-(pi(+))-nK+(pi(-)))/(nK-(pi(+))+nK+(pi(-)))=-0.133+/ 0.030(stat)+/-0.009(syst). The probability of observing such an asymmetry in the absence of direct CP violation is 1.3x10(-5), corresponding to 4.2 standard deviations. PMID- 15524720 TI - Whispering gallery modes inside asymmetric resonant cavities. AB - Two dimensional resonators with a smooth strictly convex boundary are known to possess a whispering gallery region supporting modes concentrated near the boundary. A new class of asymmetric resonant cavities is introduced, where a whispering gallery-like region is found deep inside the resonator. The construction of such resonators is a novel application of the geometric control methods. The results of numerical simulations and experiments are presented. PMID- 15524707 TI - Measurements of CP-violating asymmetries in B0-->K(0)(s)pi(0) decays. AB - We present a measurement of the time-dependent CP-violating (CPV) asymmetries in B0-->K(0)(S)pi(0) decays based on 124x10(6) Upsilon(4S)-->BB decays collected with the BABAR detector at the PEP-II asymmetric-energy B factory at SLAC. In a sample containing 122+/-16 signal decays, we obtain the magnitudes of the direct CPV asymmetry CK(0)(S)(pi(0))=0.40(+0.27)(-0.28)+/-0.09 and of the CPV asymmetry in the interference between mixing and decay SK(0)(S)(pi(0))=0.48(+0.38)(-0.47)+/ 0.06 where the first error is statistical and the second systematic. PMID- 15524723 TI - Phase synchronization in ensembles of bursting oscillators. AB - We study the effects of mutual and external chaotic phase synchronization in ensembles of bursting oscillators. These oscillators (used for modeling neuronal dynamics) are essentially multiple time scale systems. We show that a transition to mutual phase synchronization takes place on the bursting time scale of globally coupled oscillators, while on the spiking time scale, they behave asynchronously. We also demonstrate the effect of the onset of external chaotic phase synchronization of the bursting behavior in the studied ensemble by a periodic driving applied to one arbitrarily taken neuron. We also propose an explanation of the mechanism behind this effect. We infer that the demonstrated phenomenon can be used efficiently for controlling bursting activity in neural ensembles. PMID- 15524722 TI - Conical second harmonic generation in a two-dimensional chi(2) photonic crystal: a hexagonally poled LiTaO3 crystal. AB - A new type of conical second-harmonic generation was discovered in a 2D chi((2)) photonic crystal-a hexagonally poled LiTaO3 crystal. It reveals the presence of another type of nonlinear interaction-a scattering involved optical parametric generation in a nonlinear medium. Such a nonlinear interaction can be significantly enlarged in a modulated chi((2)) structure by a quasi-phase matching process. The conical beam records the spatial distribution of the scattering signal and discloses the structure information and symmetry of the 2D chi((2)) photonic crystal. PMID- 15524724 TI - First experimental evidence for quantum echoes in scattering systems. AB - A self-pulsing effect termed quantum echoes has been observed in experiments with an open superconducting and a normal conducting microwave billiard whose geometry provides soft chaos, i.e., a mixed phase space portrait with a large stable island. For such systems a periodic response to an incoming pulse has been predicted. Its period has been associated with the degree of development of a horseshoe describing the topology of the classical dynamics. The experiments confirm this picture and reveal the topological information. PMID- 15524726 TI - Lagrangian tracers on a surface flow: the role of time correlations. AB - Finite time correlations of the velocity in a surface flow are found to be important for the formation of clusters of Lagrangian tracers. The degree of clustering characterized by the Lyapunov spectrum of the flow is numerically shown to be in qualitative agreement with the predictions for the white-in-time compressible Kraichnan flow, but to deviate quantitatively. For intermediate values of compressibility the clustering is surprisingly weakened by time correlations. PMID- 15524725 TI - Transient structures in a granular gas. AB - A force-free granular gas is considered with an impact-velocity-dependent coefficient of restitution as it follows from the model of viscoelastic particles. We analyze structure formation in this system by means of three independent methods: molecular dynamics, numerical solution of the hydrodynamic equations, and linear stability analysis of these equations. All these approaches indicate that structure formation occurs in force-free granular gases only as a transient process. PMID- 15524727 TI - Dynamics in a multicomponent plasma near the low-frequency cutoff. AB - The distinctive feature of a multicomponent plasma is found to be a first order rotation of the light ion fluid with a characteristic frequency, Omega(r). A resonance at omega=Omega(r) results in plasma oscillation with unique properties. Coupling of the fast rotation time scale with the slow magnetosonic time scale leads to a nonlinear Schro dinger equation for the system and suggests the possibility of strong structural turbulence at MHD scales. PMID- 15524728 TI - Measurement of the resistive-wall-mode stability in a rotating plasma using active MHD spectroscopy. AB - The stability of the resistive-wall mode (RWM) in DIII-D plasmas above the conventional pressure limit, where toroidal plasma rotation in the order of a few percent of the Alfve n velocity is sufficient to stabilize the n=1 RWM, has been probed using the technique of active MHD spectroscopy at frequencies of a few Hertz. The measured frequency spectrum of the plasma response to externally applied rotating resonant magnetic fields is well described by a single-mode approach and provides an absolute measurement of the damping rate and the natural mode rotation frequency of the stable RWM. PMID- 15524729 TI - Parametric dependence of turbulent particle transport in tore supra plasmas. AB - Steady state full noninductive current tore supra plasmas offer a unique opportunity to study the local parametric dependence of particle pinch velocity, in order to discriminate among different theories. Magnetic field shear is found to generate an inward pinch which is dominant in the gradient region (normalized radius 0.3>1 the shear stress at t=0(+) is proportional to gamma(-1)lngamma, the first normal stress is proportional, lngamma and both the stress-optical law and the time-strain factorability are violated. A principle of virtual work applies to fast deformations. PMID- 15524737 TI - Atomic-resolution dynamic force microscopy and spectroscopy of a single-walled carbon nanotube: characterization of interatomic van der Waals forces. AB - We report atomic-resolution imaging and site-specific quantitative force measurements on a single-walled carbon nanotube by dynamic force microscopy and three-dimensional force field spectroscopy at low temperatures. The topography imaged in the attractive force regime reflects the trigonal arrangement of the hollow sites as maxima. Individual force curves were unambiguously assigned to carbon atoms and hollow sites, respectively. Site-specific quantitative evaluation revealed that the short-range interatomic van der Waals forces are responsible for the atomic-scale contrast. PMID- 15524738 TI - Rule for structures of open metal surfaces. AB - We present a clear and simple rule for determining the relaxation sequences on open (stepped, vicinal, or high-Miller-index) metal surfaces. At the bulk truncated configuration of a surface, a surface slab is defined where the coordination of atoms is reduced from the bulk. The rule predicts that the interlayer spacings within this slab contract, while the interlayer spacing between this slab and the substrate expands. By first-principles calculations, we show that this rule is obeyed on all open Cu surfaces with interlayer spacings down to about 0.5 A. We also illustrate a direct relation of the relaxation sequences to the charge redistribution on these surfaces, which is demonstrated to be driving the multilayer relaxations. The applicability of the rule can be extended to other fcc and bcc metals, including unreconstructed and missing-row surfaces. PMID- 15524739 TI - Phonon spectromicroscopy of carbon nanostructures with atomic resolution. AB - The vibrational properties of single-wall carbon nanotubes have been probed locally with atomic-scale resolution by inelastic electron tunneling spectroscopy with a low-temperature scanning tunneling microscope. The high spatial resolution has allowed the unraveling of changes in the local phonon spectrum related to topological defects. We demonstrated that the radial breathing mode is suppressed within tube segments of lengths below approximately 3 nm, and that in the cap region phonon modes characteristic of the fullerene hemisphere are emerging. Phonon spectromicroscopy should lead to a better understanding of the mechanisms that limit the transport of heat or electrical charge inside nanostructured carbon materials. PMID- 15524740 TI - Long-range surface reconstruction: Si(110)-(16 x 2). AB - A variety of reconstruction models is studied for the Si(110)-(16 x 2) surface using first-principles calculations. Assuming appropriate rebonding of edge atoms and surface chains buckled in antiphase, we show that steps along the [112] direction yielding a trench indeed lower the surface energy. We explain the long range surface reconstruction and develop a geometry model based on steps, adatoms, tetramers, and interstitials. The model is able to explain the stripes of paired pentagons seen obviously in empty-state scanning tunneling microscopy images. PMID- 15524741 TI - Formation of nanostructures by the activated Stranski-Krastanow transition method. AB - We propose that self-assembly of complicated nanocrystals can be realized by making patterns on the surface of the Stranski-Krastanow systems with the film thickness in a special range that gradual surface undulation is prohibited but island nucleation can be effected. The method has the potential to control the locations, the size, and the shapes of the self-assembled nanocrystals. PMID- 15524742 TI - Raman study of the orbital-phonon coupling in LaCoO3. AB - The magnetic state in LaCoO3 changes from the low spin state (S=0) to the mixed state with a thermally excited intermediate spin state (IS) (S=1) above about 50 K. The partially filled e(g) orbital in the IS state has a nature of Jahn-Teller (JT) distortion. The cooperative JT distortion causes an orbital order. We found that all Raman active phonon modes are affected by the excitation of IS Co3+ ions. Especially, the JT vibration mode shows anomalous temperature dependence. PMID- 15524743 TI - Angle resolved photoemission from Nd1.85Ce0.15CuO4 using high energy photons: a fermi surface investigation. AB - We have performed an angle resolved photoemission study on a single crystal of the optimally electron doped (n-type) cuprate superconductor Nd2-xCexCuO4 (x=0.15) at a photon energy of 400 eV. The Fermi surface is mapped out and is, in agreement with earlier measurements, of hole-type with the expected Luttinger volume. However, comparing with previous low energy measurements, we observe a different Fermi surface shape and a different distribution of spectral intensity around the Fermi surface contour. The observed Fermi surface shape indicates a stronger electron correlation in the bulk as compared to the surface. PMID- 15524744 TI - Quantum percolation in granular metals. AB - Theory of quantum corrections to conductivity of granular metal films is developed for the realistic case of large randomly distributed tunnel conductances. Quantum fluctuations of intergrain voltages (at energies E much below the bare charging energy scale E(C)) suppress the mean conductance g (E) much more strongly than its standard deviation sigma(E). At sufficiently low energies E(*) any distribution becomes broad, with sigma(E(*)) approximately g (E(*)), leading to strong local fluctuations of the tunneling density of states. The percolative nature of the metal-insulator transition is established by a combination of analytic and numerical analysis of the matrix renormalization group equations. PMID- 15524745 TI - Ab-initio electron transport calculations of carbon based string structures. AB - First-principles calculations show that monatomic strings of carbon have high cohesive energy and axial strength, and exhibit stability even at high temperatures. Because of their flexibility and reactivity, carbon chains are suitable for structural and chemical functionalizations; they also form stable ring, helix, grid, and network structures. Analysis of electronic conductance of various infinite, finite, and doped string structures reveal fundamental and technologically interesting features. Changes in doping and geometry give rise to dramatic variations in conductance. In even-numbered linear chains, strain induces a substantial decrease of conductance. The double covalent bonding of carbon atoms underlies their unusual chemical, mechanical, and transport properties. PMID- 15524746 TI - Projective quantum monte carlo method for the anderson impurity model and its application to dynamical mean field theory. AB - We develop a projective quantum Monte Carlo algorithm of the Hirsch-Fye type for obtaining ground state properties of the Anderson impurity model. This method is employed to solve the self-consistency equations of dynamical mean field theory. It is shown that the approach converges rapidly to the ground state so that reliable zero-temperature results are obtained. As a first application, we study the Mott-Hubbard metal-insulator transition of the frustrated one-band Hubbard model, reconfirming the numerical renormalization group results. PMID- 15524747 TI - Spin-dependent transport in molecular tunnel junctions. AB - We present measurements of magnetic tunnel junctions made using a self-assembled monolayer molecular barrier. Ni-octanethiol-Ni samples were fabricated in a nanopore geometry. The devices exhibit significant changes in resistance as the angle between the magnetic moments in the two electrodes is varied, demonstrating that low-energy electrons can traverse the molecular barrier while remaining spin polarized. An analysis of the voltage and temperature dependence of the data suggests that the spin-polarized transport signals can be degraded by localized states in the molecular barriers. PMID- 15524748 TI - Asymmetric quantum shot noise in quantum dots. AB - We analyze the frequency-dependent noise of a current through a quantum dot which is coupled to Fermi leads and which is in the Coulomb blockade regime. We show that the asymmetric shot noise, as a function of detection frequency, shows steps and becomes super-Poissonian. This provides experimental access to the quantum fluctuations of the current. We present an exact calculation of the noise for a single dot level and a perturbative evaluation of the noise in Born approximation (sequential tunneling regime but without Markov approximation) for the general case of many levels with charging interaction. PMID- 15524749 TI - Chemical tuning of metal-semiconductor interfaces. AB - We report a study of the Schottky barrier for Pb films grown on Si surfaces terminated by various metals (Ag, In, Au, and Pb) to explore the atomic-scale physics of the interface barrier and a means to control the barrier height. Electronic confinement by the Schottky barrier results in quantum well states in the Pb films, which are measured by angle-resolved photoemission. The barrier height is determined from the atomic-layer-resolved energy levels and the line widths. A calculation based on the known interface chemistry and the electronegativity yields predicted barrier heights in good agreement with the experiment. PMID- 15524750 TI - Single-electron tunneling with strong mechanical feedback. AB - A harmonic nanomechanical oscillator with a high quality factor weakly coupled to a single-electron tunneling device can provide a strong feedback for electron transport. Strong feedback occurs in a narrow voltage range just above the Coulomb blockade threshold. In this regime, current is strongly modified and current noise is drastically enhanced compared to the Schottky value. PMID- 15524751 TI - Superconducting proximity effect at the paramagnetic-ferromagnetic transition. AB - The exchange-enhanced electron-electron interactions at the paramagnetic ferromagnetic transition were studied experimentally via proximity effect tunneling spectroscopy. By solving the Usadel equations in both the paramagnetic and ferromagnetic states, the electron-spin fluctuation coupling constant and the exchange field are derived from the tunneling spectra. PMID- 15524752 TI - Magneto-optical evidence for a gapped fermi surface in underdoped YBa2Cu3O6+x. AB - The infrared (900-1100 cm(-1)) Faraday rotation and circular dichroism are measured in the normal state of underdoped High T(c) superconductors and used to study the magnetotransport. YBa2Cu3O6+x thin films are investigated in the temperature range 10-300 K in magnetic fields up to 8 T and as a function of oxygen concentration. A dramatic increase of the Hall frequency is observed for underdoped samples, which is not consistent with the approach to a Mott transition but is consistent with a partial gapping of the Fermi surface as predicted in density wave models. PMID- 15524754 TI - Magnetic percolation in diluted magnetic semiconductors. AB - We demonstrate that the magnetic properties of diluted magnetic semiconductors are dominated by short ranged interatomic exchange interactions that have a strong directional dependence. By combining first principles calculations of interatomic exchange interactions with a classical Heisenberg model and Monte Carlo simulations, we reproduce the observed critical temperatures of a broad range of diluted magnetic semiconductors. We also show that agreement between theory and experiment is obtained only when the magnetic atoms are randomly positioned. This suggests that the ordering of diluted magnetic semiconductors is heavily influenced by magnetic percolation, and that the measured critical temperatures should be very sensitive to details in the sample preparation, in agreement with observations. PMID- 15524753 TI - Pressure enhancement of the giant magnetocaloric effect in Tb5Si2Ge2. AB - Effects of temperature and pressure on magnetic, elastic, structural, and thermal properties of Tb5Si2Ge2 have been studied by means of macroscopic (thermal expansion and magnetization) and microscopic (neutron powder diffraction) techniques. We present evidence that the high-temperature second-order ferromagnetic transition can be coupled with the low-temperature first-order structural phase change into a single first-order magnetic-crystallographic transformation at and above a tricritical point in the vicinity of 8.6 kbar. This pressure-induced coupling has a remarkable effect on the magnetocaloric effect, transforming Tb5Si2Ge2 from an ordinary into a giant magnetocaloric effect material. PMID- 15524755 TI - Determination of the exciton formation in quantum wells from time-resolved interband luminescence. AB - We present the results of a detailed time-resolved luminescence study carried out on a very high quality InGaAs quantum well sample where the contributions at the energy of the exciton and at the band edge can be clearly separated. We perform this experiment with a spectral resolution and a sensitivity of the setup, allowing us to keep the observation of these two separate contributions over a broad range of times and densities. This allows us to directly evidence the exciton formation time, which depends on the density as expected from theory. We also denote the dominant contribution of excitons to the luminescence signal, and the lack of thermodynamical equilibrium at low densities. PMID- 15524756 TI - Resonant enhancement of inelastic light scattering in strongly correlated materials. AB - We use dynamical mean field theory to find an exact solution for inelastic light scattering in strongly correlated materials such as those near a quantum-critical metal-insulator transition. We evaluate the results for q=0 (Raman) scattering and find that resonant effects can be quite large, and yield a double resonance, a significant enhancement of nonresonant scattering peaks, a joint resonance of both peaks when the incident photon frequency is on the order of U, and the appearance of an isosbestic point in all symmetry channels for an intermediate range of incident photon frequencies. PMID- 15524757 TI - Fullerene quantum gyroscope. AB - We report the observation of quantized rotational states of a diatomic C2 unit in solid endohedral fullerene C(2)Sc(2)@C(84). The rotational transitions induce a periodic line pattern in the low energy Raman spectrum. The rotational constant B and the C-C distance were found to be 1.73 cm(-1) and 0.127 nm, respectively. Density functional calculations revealed an intrinsic rotational barrier of the order of only a few meV for the C2 unit. The Schrodinger equation involving the potential barrier was solved and the Raman tensor matrix elements were calculated, yielding good quantitative agreement with the experiment. To our best knowledge this is the first intrinsic rotational spectrum of a diatomic plane molecular rotor. PMID- 15524758 TI - Nanofocusing of optical energy in tapered plasmonic waveguides. AB - We predict theoretically that surface plasmon polaritons propagating toward the tip of a tapered plasmonic waveguide are slowed down and asymptotically stopped when they tend to the tip, never actually reaching it (the travel time to the tip is logarithmically divergent). This phenomenon causes accumulation of energy and giant local fields at the tip. There are various prospective applications in nano optics and nanotechnology. PMID- 15524759 TI - Enhanced diffraction from a grating on the surface of a negative-index metamaterial. AB - We show by numerical simulation as well as by measurements on negative-index metamaterial wedge samples, that the unavoidable stepping of the refraction interface-due to the finite unit-cell size inherent to metamaterials-can give rise to a well-defined diffracted beam in addition to the negatively refracted beam. The direction of the diffracted beam is consistent with elementary diffraction theory; however, the coupling to this higher order beam is much larger than would be the case for a positive index material. The results confirm recent theoretical predictions of enhanced diffraction for negative-index grating surfaces. PMID- 15524760 TI - Kinetic electron excitation in atomic collision cascades. AB - The kinetic excitation of electrons upon bombardment of a solid surface with energetic ions is investigated. Using a metal-insulator-metal junction, hot electrons produced by the projectile impact are detected with excitation energies well below the vacuum level. The results provide information that cannot be accessed by electron emission experiments. The observed tunneling current depends on the projectile energy and the bias voltage across the junction, opening the possibility of internal excitation spectroscopy. PMID- 15524761 TI - Tuning the surface state dimensionality of Cu nanostripes. AB - Stepped Cu nanostripes with varying terrace widths are self-assembled during Ag induced periodic faceting of vicinal Cu(111). By changing Ag coverage the average terrace size within individual Cu stripes is readily tuned, making it possible to select the one-dimensional or two-dimensional character of surface states. Furthermore, the average terrace size can be smoothly switched from 10 to 30 A, thereby tracking the transition from step-lattice, quasi-two-dimensional umklapp bands to terrace-confined one-dimensional quantum well states. PMID- 15524762 TI - Time resolved experimental analysis of the electric field induced biaxial order reconstruction in nematics. AB - We present the time resolved experimental characterization of the biaxial switching between two topologically distinct textures of a nematic liquid crystal cell submitted to a strong electric field. This fast electro-optical effect is governed by the electric induced order reconstruction in the nematic bulk, which is an actual challenge for a complete theoretical description of the nematodynamics. The electric measurements across the cell are suitable to this purpose since they well discriminate among the dielectric, the ionic, and the order reconstruction contributions. A phenomenological model describes the experimental data, allowing the measurements of the order reconstruction characteristic time. PMID- 15524763 TI - Peeling, healing, and bursting in a lubricated elastic sheet. AB - We consider the dynamics of an elastic sheet lubricated by the flow of a thin layer of fluid that separates it from a rigid wall. By considering long wavelength deformations of the sheet, we derive an evolution equation for its motion, accounting for the effects of elastic bending, viscous lubrication, and body forces. We then analyze various steady and unsteady problems for the sheet, such as peeling, healing, levitating, and bursting, using a combination of numerical simulation and dimensional analysis. On the macroscale, we corroborate our theory with a simple experiment, and, on the microscale, we analyze an oscillatory valve that can transform a continuous stream of fluid into a series of discrete pulses. PMID- 15524764 TI - Attractive instability of oppositely charged membranes induced by charge density fluctuations. AB - We predict the conditions under which two oppositely charged membranes show a dynamic, attractive instability. Two layers with unequal charges of opposite sign can repel or be stable when in close proximity. However, dynamic charge density fluctuations can induce an attractive instability and thus facilitate fusion. We predict the dominant instability modes and time scales and show how these are controlled by the relative charge and membrane viscosities. These dynamic instabilities may be the precursors of membrane fusion in systems where artificial vesicles are engulfed by biological cells of opposite charge. PMID- 15524765 TI - Liouville dynamics and the conservation of classical information. AB - We show that the invariance of entropic distances under Liouville dynamics yields classical analogues of information-related, quantum mechanical impossible operations. By recourse to Fisher's information measure, we also establish a connection between such classically forbidden operations and statistical estimation theory. PMID- 15524766 TI - Comment on "collective excitations of a degenerate gas at the BEC-BCS crossover". PMID- 15524767 TI - Comment on "memory effects in an interacting magnetic nanoparticle system". PMID- 15524768 TI - Comment on "memory effects in an interacting magnetic nanoparticle system". PMID- 15524770 TI - Coherent classical-path description of deep tunneling. AB - A central challenge to the semiclassical description of quantum mechanics is the quantum phenomenon of "deep" tunneling. Here we show that real time classical trajectories suffice to account correctly even for deep quantum tunneling, using a recently formulated semiclassical initial value representation series of the quantum propagator and a prefactor free semiclassical propagator. Deep quantum tunneling is effected through what we term as coherent classical paths which are composed of one or more classical trajectories that lead from reactant to product but are discontinuous along the way. The end and initial phase space points of consecutive classical trajectories contributing to the coherent path are close to each other in the sense that the distance between them is weighted by a coherent state overlap matrix element. Results are presented for thermal and energy dependent tunneling through a symmetric Eckart barrier. PMID- 15524771 TI - Locality in quantum and Markov dynamics on lattices and networks. AB - We consider gapped systems governed by either quantum or Markov dynamics, with the low-lying states below the gap being approximately degenerate. For a broad class of dynamics, we prove that ground or stationary state correlation functions can be written as a piece decaying exponentially in space plus a term set by matrix elements between the low-lying states. The key to the proof is a local approximation to the negative energy, or annihilation, part of an operator in a gapped system. Applications to numerical simulation of quantum systems and to networks are discussed. PMID- 15524772 TI - The second law, Maxwell's demon, and work derivable from quantum heat engines. AB - With a class of quantum heat engines which consists of two-energy-eigenstate systems undergoing, respectively, quantum adiabatic processes and energy exchanges with heat baths at different stages of a cycle, we are able to clarify some important aspects of the second law of thermodynamics. The quantum heat engines also offer a practical way, as an alternative to Szilard's engine, to physically realize Maxwell's demon. While respecting the second law on the average, they are also capable of extracting more work from the heat baths than is otherwise possible in thermal equilibrium. PMID- 15524773 TI - Finite-time disentanglement via spontaneous emission. AB - We show that under the influence of pure vacuum noise two entangled qubits become completely disentangled in a finite-time, and in a specific example we find the time to be given by ln((2+sqrt[2] / 2) times the usual spontaneous lifetime. PMID- 15524774 TI - Molecular matter-wave amplifier. AB - We describe a matter-wave amplifier for vibrational ground-state molecules which uses a Feshbach resonance to first form quasibound molecules starting from an atomic Bose-Einstein condensate. The quasibound molecules are then driven into their stable vibrational ground state via a two-photon Raman transition inside an optical cavity. The transition from the quasibound state to the electronically excited state is driven by a classical field. Amplification of ground state molecules is then achieved by using a strongly damped cavity mode for the transition from the electronically excited molecules to the molecular ground state. PMID- 15524775 TI - Observation of dynamical instability for a Bose-Einstein condensate in a moving 1D optical lattice. AB - We have experimentally studied the unstable dynamics of a harmonically trapped Bose-Einstein condensate loaded into a 1D moving optical lattice. The lifetime of the condensate in such a potential exhibits a dramatic dependence on the quasimomentum state. This is unambiguously attributed to the onset of dynamical instability, after a comparison with the predictions of the Gross-Pitaevskii theory. Deeply in the unstable region we observe the rapid appearance of complex structures in the atomic density profile, as a consequence of the condensate phase uniformity breakdown. PMID- 15524776 TI - Gisin's theorem for three qubits. AB - We present a theorem that all generalized Greenberger-Horne-Zeilinger states of a three-qubit system violate a Bell inequality in terms of probabilities. All pure entangled states of a three-qubit system are shown to violate a Bell inequality for probabilities; thus, one has Gisin's theorem for three qubits. PMID- 15524777 TI - Single atom transistor in a 1D optical lattice. AB - We propose a scheme utilizing a quantum interference phenomenon to switch the transport of atoms in a 1D optical lattice through a site containing an impurity atom. The impurity represents a qubit which in one spin state is transparent to the probe atoms, but in the other acts as a single atom mirror. This allows a single-shot quantum nondemolition measurement of the qubit spin. PMID- 15524778 TI - Universal quantum computation through control of spin-orbit coupling. AB - We propose a method for quantum computation which uses control of spin-orbit coupling in a linear array of single electron quantum dots. Quantum gates are carried out by pulsing the exchange interaction between neighboring electron spins, including the anisotropic corrections due to spin-orbit coupling. Control over these corrections, even if limited, is sufficient for universal quantum computation over qubits encoded into pairs of electron spins. The number of voltage pulses required to carry out either single-qubit rotations or controlled Not gates scales as the inverse of a dimensionless measure of the degree of control of spin-orbit coupling. PMID- 15524779 TI - Measurement of gravitational spin-orbit coupling in a binary-pulsar system. AB - In relativistic gravity, a spinning pulsar will precess as it orbits a compact companion star. We have measured the effect of such precession on the average shape and polarization of the radiation from PSR B1534+12. We have also detected, with limited precision, special-relativistic aberration of the revolving pulsar beam due to orbital motion. Our observations fix the system geometry, including the misalignment between the spin and orbital angular momenta, and yield a measurement of the precession time scale consistent with the predictions of general relativity. PMID- 15524783 TI - Prethermalization. AB - Prethermalization of the equation of state and the kinetic temperature to their equilibrium values occurs on time scales dramatically shorter than the thermal equilibration time. This is a crucial ingredient for the understanding of collisions of heavy nuclei or other nonequilibrium phenomena in complex quantum and classical many body systems. We also compare the chemical equilibration time with other characteristic time scales. PMID- 15524784 TI - Laser spectroscopic determination of the 6He nuclear charge radius. AB - We have performed precision laser spectroscopy on individual 6He (t(1/2)=0.8 s) atoms confined and cooled in a magneto-optical trap, and measured the isotope shift between 6He and 4He to be 43 194.772+/-0.056 MHz for the 2(3)S1-3(3)P2 transition. Based on this measurement and atomic theory, the nuclear charge radius of 6He is determined for the first time in a method independent of nuclear models to be 2.054+/-0.014 fm. The result is compared with the values predicted by a number of nuclear structure calculations and tests their ability to characterize this loosely bound halo nucleus. PMID- 15524785 TI - Effective-range approach and scaling laws for electromagnetic strength in neutron halo nuclei. AB - We study low-lying multipole strength in neutron-halo nuclei. The strength depends only on a few low-energy constants: the neutron separation energy, the asymptotic normalization coefficient of the bound-state wave function, and the scattering length that contains the information on the interaction in the continuum. The shape of the transition probability shows a characteristic dependence on few scaling parameters and the angular momenta. The total E1 strength is related to the root-mean-square radius of the neutron wave function in the ground state and shows corresponding scaling properties. We apply our approach to the E1 strength distribution of 11Be. PMID- 15524786 TI - Magnetic moment of the fragmentation-aligned 61Fe (9/2(+)) isomer. AB - We report on the g factor measurement of an isomer in the neutron-rich (61)(26)Fe (E(*)=861 keV and T(1/2)=239(5) ns). The isomer was produced and spin aligned via a projectile-fragmentation reaction at intermediate energy, the time dependent perturbed angular distribution method being used for the measurement of the g factor. For the first time, due to significant improvements of the experimental technique, an appreciable residual alignment of the nuclear spin ensemble has been observed, allowing a precise determination of its g factor, including the sign: g=-0.229(2). In this way we open the possibility to study moments of very neutron-rich short-lived isomers, not accessible via other production and spin orientation methods. PMID- 15524787 TI - Large enhancement of radiative strength for soft transitions in the quasicontinuum. AB - Radiative strength functions (RSFs) for the (56,57)Fe nuclei below the separation energy are obtained from the 57Fe(3He,alphagamma)56Fe and 57Fe(3He,3He'gamma)57Fe reactions, respectively. An enhancement of more than a factor of 10 over common theoretical models of the soft (E(gamma) less than or approximately equal 2 MeV) RSF for transitions in the quasicontinuum (several MeV above the yrast line) is observed. Two-step cascade intensities with soft primary transitions from the 56Fe(n,2gamma)57Fe reaction confirm the enhancement. PMID- 15524788 TI - Observation of Feshbach resonances between two different atomic species. AB - We have observed three Feshbach resonances in collisions between 6Li and 23Na atoms. The resonances were identified as narrow loss features when the magnetic field was varied. The molecular states causing these resonances have been identified, and additional 6Li-23Na resonances are predicted. These resonances will allow the study of degenerate Bose-Fermi mixtures with adjustable interactions and could be used to generate ultracold heteronuclear molecules. PMID- 15524789 TI - Three-boson problem near a narrow Feshbach resonance. AB - We consider a three-boson system with resonant binary interactions and show that for sufficiently narrow resonances three-body observables depend only on the resonance width and the scattering length. The effect of narrow resonances is qualitatively different from that of wide resonances revealing novel physics of three-body collisions. We calculate the rate of three-body recombination to a weakly bound level and the atom-dimer scattering length and discuss implications for experiments on Bose-Einstein condensates and atom-molecule mixtures near Feshbach resonances. PMID- 15524790 TI - Determination of the number of atoms trapped in an optical cavity. AB - The number of atoms trapped within the mode of an optical cavity is determined in real time by monitoring the transmission of a weak probe beam. Continuous observation of atom number is accomplished in the strong coupling regime of cavity quantum electrodynamics and functions in concert with a cooling scheme for radial atomic motion. The probe transmission exhibits sudden steps from one plateau to the next in response to the time evolution of the intracavity atom number, from N>or=3 to N=2-->1-->0 atoms, with some trapping events lasting over 1 s. PMID- 15524791 TI - Generalized sagnac effect. AB - Experiments were conducted to study light propagation in a light waveguide loop consisting of linearly and circularly moving segments. We found that any segment of the loop contributes to the total phase difference between two counterpropagating light beams in the loop. The contribution is proportional to a product of the moving velocity v and the projection of the segment length Delta(l) on the moving direction, Deltaphi=4piv x Delta(l)/c(lambda). It is independent of the type of motion and the refractive index of waveguides. The finding includes the Sagnac effect of rotation as a special case and suggests a new fiber optic sensor for measuring linear motion with nanoscale sensitivity. PMID- 15524792 TI - Soliton eigenvalue control in optical lattices. AB - We address the dynamics of higher-order solitons in optical lattices, and predict their self-splitting into the set of their single-soliton constituents. The splitting is induced by the potential introduced by the lattice, together with the imprinting of a phase tilt onto the initial multisoliton states. The phenomenon allows the controllable generation of several coherent solitons linked via their Zakharov-Shabat eigenvalues. Application of the scheme to the generation of correlated matter waves in Bose-Einstein condensates is discussed. PMID- 15524793 TI - Field inside a random distribution of parallel dipoles. AB - We determine the probability distribution for the field inside a random distribution of electric or magnetic dipoles. Although the average contribution from any spherical shell around the probe position vanishes, at the center of a spherical distribution of parallel dipoles, the Levy stable distribution of the field is symmetric around a nonvanishing field amplitude. Omission of contributions from a small volume around the probe leads to a field distribution with a vanishing mean, which, in the limit of vanishing excluded volume, converges to the shifted distribution. PMID- 15524794 TI - Ultraslow optical solitons in a cold four-state medium. AB - We show the formation of ultraslow optical solitons in a lifetime broadened four state atomic medium under Raman excitation. With appropriate conditions we demonstrate, both analytically and numerically, that both bright and dark ultraslow optical solitons can occur in such a highly resonant medium with remarkable propagation characteristics. This work may open other research opportunities in condensed matter and may result in a substantial impact on technology. PMID- 15524795 TI - Control of the refractive index in photopolymerizable materials for (2+1)D solitary wave guide formation. AB - We report an experimental and theoretical study on the optimization of (2+1)D self-written waveguide formation inside a photopolymerizable material. The accurate control of the refractive index value inside the bulk of the material during the polymerization process gives us the opportunity to define a virtual core and a virtual cladding for the system. The V value which characterizes the guidance properties of a fiber can be applied to this propagation. The control of the V value allows us to propagate single mode or multimode waveguides on a few centimeters. Numerical simulations of these waveguides based on a paraxial model including both photopolymerization and Kerr effect give very good agreement with our experimental results. PMID- 15524796 TI - Magnetic field enhanced coherence length in cold atomic gases. AB - We study the effect of an external magnetic field on coherent backscattering of light from a cold rubidium vapor. We observe that the backscattering enhancement factor can be increased with B. This surprising behavior shows that the coherence length of the system can be increased by adding a magnetic field, in sharp contrast with usual situations. This is mainly due to the lifting of the degeneracy between Zeeman sublevels. We find good agreement between our experimental data and a full Monte Carlo simulation, taking into account the magneto-optical effects and the geometry of the atomic cloud. PMID- 15524797 TI - Effective Kerr nonlinearity and two-color solitons in photonic band-gap fibers filled with a Raman active gas. AB - We predict a strong effective Kerr nonlinearity in hollow-core photonic crystal fibers filled with a Raman active gas which exceeds the intrinsic Kerr nonlinearity by 2 orders of magnitude. Two-color bright-bright and dark-bright solitons supported by this nonlinearity are found and the feasibility of their experimental observation is demonstrated. PMID- 15524798 TI - Experimental deterministic coherence resonance. AB - We demonstrate coherence resonance in a dynamical system without external noise. The experimental evidence is reported in the low frequency fluctuations of a chaotic diode laser with optical feedback. The phenomenon is also verified numerically using the Lang-Kobayashi equations for a single solitary mode laser, without noise terms. Fast deterministic dynamics plays the role of an effective exciting noise, narrowing the resonance in the autonomous slow power drop cycles of the laser. This new result is the natural extension of deterministic stochastic resonance and noise induced coherence resonance predicted and observed in recent years. PMID- 15524781 TI - Search for doubly charged higgs boson pair production in the decay to mu(+)mu(+)mu(-)mu(-) in pp collisions at sqrt[s]=1.96 TeV. AB - A search for pair production of doubly charged Higgs bosons in the process pp - >H(++)H(--) -->mu(+)mu(+)mu(-)mu(-) is performed with the D0 run II detector at the Fermilab Tevatron. The analysis is based on a sample of inclusive dimuon data collected at an energy of sqrt[s]=1.96 TeV, corresponding to an integrated luminosity of 113 pb(-1). In the absence of a signal, 95% confidence level mass limits of M(H(+/-+/-)(L))>118.4 GeV/c(2) and M(H(+/-+/-)(R))>98.2 GeV/c(2) are set for left-handed and right-handed doubly charged Higgs bosons, respectively, assuming 100% branching into muon pairs. PMID- 15524799 TI - Model for heat conduction in nanofluids. AB - A comprehensive model has been proposed to account for the large enhancement of thermal conductivity in nanofluids and its strong temperature dependence, which the classical Maxwellian theory has been unable to explain. The dependence of thermal conductivity on particle size, concentration, and temperature has been taken care of simultaneously in our treatment. While the geometrical effect of an increase in surface area with a decrease in particle size, rationalized using a stationary particle model, accounts for the conductivity enhancement, a moving particle model developed from the Stokes-Einstein formula explains the temperature effect. Predictions from the combined model agree with the experimentally observed values of conductivity enhancement of nanofluids. PMID- 15524780 TI - Inclusive double-pomeron exchange at the fermilab tevatron p p collider. AB - We report results from a study of events with a double-Pomeron exchange topology produced in p p collisions at sqrt[s]=1800 GeV. The events are characterized by a leading antiproton and a large rapidity gap on the outgoing proton side. We find that the differential production cross section agrees in shape with predictions based on Regge theory and factorization, and that the ratio of double-Pomeron exchange to single diffractive production rates is relatively unsuppressed as compared to the O(10) suppression factor previously measured in single diffractive production. PMID- 15524800 TI - Falling paper: Navier-Stokes solutions, model of fluid forces, and center of mass elevation. AB - We investigate the problem of falling paper by solving the two dimensional Navier Stokes equations subject to the motion of a free-falling body at Reynolds numbers around 10(3). The aerodynamic lift on a tumbling plate is found to be dominated by the product of linear and angular velocities rather than velocity squared, as appropriate for an airfoil. This coupling between translation and rotation provides a mechanism for a brief elevation of center of mass near the cusplike turning points. The Navier-Stokes solutions further provide the missing quantity in the classical theory of lift, the instantaneous circulation, and suggest a revised model for the fluid forces. PMID- 15524801 TI - Convective roll dynamics in liquid 4He near the onset of convection. AB - We present results of experiments on Rayleigh-Benard convection in liquid 4He at several temperatures. We show visually that with carefully defined boundary conditions the basic convection state consists of parallel rolls which are aligned in one of two directions, the angle thus defined as being temperature dependent and we attempt to explain this behavior. We also show directly the skew varicose instability acting on the basic state and correlate it with fluctuations in the temperature difference across the fluid layer. PMID- 15524782 TI - Measurement of the tt production cross section in pp collisions at sqrt[s]=1.96 TeV using dilepton events. AB - We report a measurement of the tt production cross section using dilepton events with jets and missing transverse energy in pp collisions at a center-of-mass energy of 1.96 TeV. Using a 197+/-12 pb(-1) data sample recorded by the upgraded Collider Detector at Fermilab, we use two complementary techniques to select candidate events. We compare the number of observed events and selected kinematical distributions with the predictions of the standard model and find good agreement. The combined result of the two techniques yields a tt production cross section of 7.0(+2.4)(-2.1)(stat)+1.6-1.1(syst)+/-0.4(lum) pb. PMID- 15524803 TI - Mass-profile and instability-growth measurements for 300-wire Z-pinch implosions driven by 14-18 MA. AB - We present the first comprehensive study of high wire-number, wire-array Z-pinch dynamics at 14-18 MA using x-ray backlighting and optical shadowgraphy diagnostics. The cylindrical arrays retain slowly expanding, dense wire cores at the initial position up to 60% of the total implosion time. Azimuthally correlated instabilities at the array edge appear during this stage which continue to grow in amplitude and wavelength after the start of bulk motion, resulting in measurable trailing mass that does not arrive on axis before peak x ray emission. PMID- 15524802 TI - Particle-transport analysis in reversed field pinch helical states. AB - This Letter reports the result of a numerical study of particle transport in self organized single helicity (SH) and quasi-SH reversed field pinch plasmas. Our code, benchmarked against experimental data, predicts a large improvement in particle transport for SH compared to the standard multiple helicity states. The contribution of neoclassical effects is noted. An estimate of the ambipolar electric field in helical states and in fully stochastic magnetic fields is given. PMID- 15524804 TI - Trapped-particle instability leading to bursting in stimulated Raman scattering simulations. AB - Nonlinear, kinetic simulations of stimulated Raman scattering (SRS) under laser fusion conditions present a bursting behavior. Different explanations for this regime have been given in previous studies: saturation of SRS by increased nonlinear Landau damping [K. Estabrook et al., Phys. Fluids B 1, 1282 (1989)]], and detuning due to the nonlinear frequency shift of the plasma wave [H. X. Vu et al., Phys. Rev. Lett. 86, 4306 (2001)]]. Another mechanism, also assigning a key role to the trapped electrons is proposed here: the breakup of the plasma wave through the trapped-particle instability. PMID- 15524805 TI - Fast-ion production from short-pulse irradiation of ethanol microdroplets. AB - We report on the first detailed measurements of ion energy distributions from microdroplet targets under ultrashort (100 fs), intense (10(16) Wcm(-2)) laser irradiation. Protons with kinetic energies of up to 20 keV are produced in a strongly anisotropic microexplosion. Modeling of the incident electromagnetic field indicates strong spatial variations over the surface of the microdroplet. Detailed numerical modeling shows that nonuniform heating on a submicron scale causes temperature inhomogeneities and nonradial pressure gradients, which drive an energetic nonspherical expansion. PMID- 15524806 TI - Specific heat of disordered superfluid 3He. AB - The specific heat of superfluid 3He, disordered by a silica aerogel, is found to have a sharp discontinuity marking the thermodynamic transition to superfluidity at a temperature reduced from that of bulk 3He. The magnitude of the discontinuity is also suppressed. This disorder effect can be understood from the Ginzburg-Landau theory which takes into account elastic quasiparticle scattering suppressing both the transition temperature and the amplitude of the order parameter. We infer that the limiting temperature dependence of the specific heat is linear at low temperatures in the disordered superfluid state, consistent with predictions of gapless excitations everywhere on the Fermi surface. PMID- 15524807 TI - A1 and A2 transitions in superfluid 3He in 98% porosity aerogel. AB - Superfluid 3He in high porosity aerogel is the system in which the effects of static impurities on a p-wave superfluid can be investigated in a systematic manner. We performed shear acoustic impedance measurements on this system (98% porosity aerogel) in the presence of magnetic fields up to 15 T at the sample pressures of 28.4 and 33.5 bars. We observed the splitting of the superfluid transition into two transitions in high fields in both bulk and liquid in aerogel. The field dependence of the splitting in aerogel resembles that of the bulk superfluid 3He caused by the presence and growth of the A1 phase. Our results provide the first evidence of the A1 phase in superfluid (3)He/aerogel. PMID- 15524808 TI - Excitations of metastable superfluid 4He at pressures up to 40 bars. AB - Neutron scattering measurements of the fundamental excitations of liquid 4He confined in 44 A pore diameter gelsil glass at pressures up to 40 bars in the wave vector range 0.41.6 A(-1), especially the rotons, are observed up to complete solidification of all the liquid at a pressure of approximately 40 bars where the roton vanishes. At and above a pressure of 35.1 bars, Bragg peaks are observed, indicating coexistence of liquid and solid in the pores at pressures 35 less than or approximately equal P less than or approximately equal 40 bars. PMID- 15524809 TI - Structural stability of complex hydrides: LiBH4 revisited. AB - A systematic approach to study the phase stability of LiBH4 based on ab initio calculations is presented. Three thermodynamically stable phases are identified and a new phase of Cc symmetry is proposed for the first time for a complex hydride. The x-ray diffraction pattern and vibrational spectra of the Cc structure agree well with recently reported experimental data on LiBH4. Calculations of the free energy at finite temperatures suggest that the experimentally proposed P6(3)mc phase is unstable at T>0 K. PMID- 15524810 TI - Crystal-like nature of acoustic excitations in glassy ethanol. AB - We report on inelastic x-ray scattering experiments on crystalline and glassy phases of ethanol in order to directly compare the influence of disorder on high frequency acoustic excitations. We find that both the dispersion and the line width of the longitudinal acoustic excitations in the glass are the same as in the polycrystal in the reciprocal space portion covering the 1st and 2nd Brillouin zones. The structural disorder is found to play little role apart from an intrinsic angular averaging, and the nature of these excitations must essentially be the same in both glass and poly crystal. PMID- 15524811 TI - Symmetric-to-asymmetric transition in triblock copolymer-homopolymer blends. AB - In blends of a symmetric poly(styrene-ethylene/butylene-styrene) tri-block copolymer with a polystyrene homopolymer, small-angle x-ray scattering and cryotransmission electron microscopy measurements reveal a microstructure consisting of a disordered arrangement of poly(ethylene/butylene) membranes suspended in polystyrene. For triblock volume fractions less than 0.22, the membranes form an asymmetric sponge or L4 phase, consisting predominantly of equilibrium vesicles. For volume fractions greater than 0.22, they form a symmetric sponge-phase (L3 phase), separated from the L4 phase by a first-order transition. PMID- 15524812 TI - Nanopipettes for metal transport. AB - Here we demonstrate, for the first time experimentally, a nanopipette action for metals using multiwalled carbon nanotubes. The process relies on electromigration forces, created at high electron current densities, enabling the transport of material inside the hollow core of carbon nanotubes. In this way nanoparticles of iron were transported to and from electrically conducting substrates. PMID- 15524813 TI - Resonant oscillators with carbon-nanotube torsion springs. AB - We report on the characterization of nanometer-scale resonators. Each device incorporates one multiwalled carbon nanotube (MWNT) as a torsional spring. The devices are actuated electrostatically, and their deflections, both low frequency and on resonance, are detected optically. These are some of the smallest electromechanical devices ever created and are a demonstration of practical integrated MWNT-based oscillators. The results also show surprising intershell mechanical coupling behavior in the MWNTs. PMID- 15524814 TI - Atomic structure of the GaAs(001)-c(4x4) surface: first-principles evidence for diversity of heterodimer motifs. AB - The GaAs(001)-c(4x4) surface was studied using ab initio atomistic thermodynamics based on density-functional theory calculations. We demonstrate that in a range of stoichiometries, between those of the conventional three As-dimer and the new three Ga-As-dimer models, there exists a diversity of atomic structures featuring Ga-As heterodimers. These results fully explain the experimental scanning tunneling microscopy images and are likely to be relevant also to the c(4x4) reconstructed (001) surfaces of other III-V semiconductors. PMID- 15524815 TI - When Langmuir is too simple: H2 dissociation on Pd(111) at high coverage. AB - Recent experiments of H2 adsorption on Pd(111) [Nature (London) 422, 705 (2003)]] have questioned the classical Langmuir picture of second order adsorption kinetics at high surface coverage requiring pairs of empty sites for the dissociative chemisorption. Experiments find that at least three empty sites are needed. Through density functional theory, we find that H2 dissociation is favored on ensembles of sites that involve a Pd atom with no direct interaction with adsorbed hydrogen. Such active sites are formed by aggregation of at least 3 H-free sites revealing the complex structure of the "active sites." PMID- 15524816 TI - Oxidation of Pt(110). AB - Using scanning tunneling microscopy and temperature programmed desorption we investigate the Pt(110) surface under strongly oxidizing conditions involving either high-pressure O2 or atomic oxygen exposure. At low temperatures, only disordered Pt oxide structures are observed. After annealing ordered surface oxide islands are observed to coexist with a highly stable reconstructed (12x2)-O chemisorption structure. From density functional theory calculations a model for the surface oxide phase is revealed. The phase is found to be metastable, and its presence is explained in terms of stabilizing defects in the chemisorption layer and reduced Pt mobility. PMID- 15524817 TI - Mechanism of hopping transport in disordered mott insulators. AB - By using a combination of detailed experimental studies and simple theoretical arguments, we identify a novel mechanism characterizing the hopping transport in the Mott insulating phase of Ca2-xSrxRuO4 near the metal-insulator transition. The hopping exponent alpha shows a systematic evolution from a value of alpha=1/2 deeper in the insulator to the conventional Mott value alpha=1/3 closer to the transition. This behavior, which we argue to be a universal feature of disordered Mott systems close to the metal-insulator transition, is shown to reflect the gradual emergence of disorder-induced localized electronic states populating the Mott-Hubbard gap. PMID- 15524818 TI - Coupled electron-ion monte carlo calculations of dense metallic hydrogen. AB - We present an efficient new Monte Carlo method which couples path integrals for finite temperature protons with quantum Monte Carlo calculations for ground state electrons, and we apply it to metallic hydrogen for pressures beyond molecular dissociation. We report data for the equation of state for temperatures across the melting of the proton crystal. Our data exhibit more structure and higher melting temperatures of the proton crystal than do Car-Parrinello molecular dynamics results. This method fills the gap between high temperature electron proton path integral and ground state diffusion Monte Carlo methods and should have wide applicability. PMID- 15524819 TI - Resonant states in the electronic structure of the high performance thermoelectrics AgPbmSbTe2+m: the role of Ag-Sb microstructures. AB - Ab initio electronic structure calculations based on gradient corrected density functional theory were performed on a class of novel quaternary compounds AgPb(m)SbTe(2+m), which were found to be excellent high temperature thermoelctrics with a large figure of merit ZT approximately 2.2 at 800 K. We find that resonant states appear near the top of the valence and bottom of the conduction bands of bulk PbTe when Ag and Sb replace Pb. These states can be understood in terms of modified Te-Ag(Sb) bonds. The electronic structure near the gap depends sensitively on the microstructural arrangements of Ag-Sb atoms, suggesting that large ZT values may originate from the nature of these ordering arrangements. PMID- 15524820 TI - Charge and orbital order in Fe3O4. AB - Charge and orbital ordering in the low-temperature monoclinic structure of magnetite (Fe3O4) is investigated using the local spin density approximation with Coulomb interaction correction method. While the difference between t(2g) minority occupancies of Fe(2+)(B) and Fe(3+)(B) cations is large and gives direct evidence for charge ordering, the screening is so effective that the total 3d charge disproportion is rather small. The charge order has a pronounced [001] modulation, which is incompatible with the Anderson criterion. The orbital order agrees with the Kugel-Khomskii theory. PMID- 15524821 TI - Electronic correlations in oligo-acene and -thiopene organic molecular crystals. AB - From first-principles calculations we determine the Coulomb interaction between two holes on oligo-acene and -thiophene molecules in a crystal, as a function of the oligomer length. The electronic polarization of the molecules that surround the charged oligomer reduces the bare Coulomb repulsion between the holes by approximately a factor of 2. The effects of relaxing the molecular geometry in the presence of holes is found to be significantly smaller. In all cases the effective hole-hole repulsion is much larger than the valence bandwidth, which implies that at high doping levels the properties of these organic semiconductors are determined by electron-electron correlations. PMID- 15524822 TI - Electron-vibron coupling in high-resolution X-Ray absorption spectra of organic materials: NTCDA on Ag(111). AB - We report additional rich fine structures in high-resolution near-edge x-ray absorption fine structure (NEXAFS) spectra of large organic molecules using NTCDA on Ag(111) as an example. These fine structures are completely interpreted as vibronic coupling to electronic core excitations. The coupling is mode selective; predominantly one vibronic mode couples to each excitation. The fit results suggest the occurrence of a Davydov splitting, first observed for core excitons. Morphological differences substantially influence the electron-vibron coupling, indicating a strong intermolecular interaction. Thus NEXAFS becomes a more subtle probe for organic solids. PMID- 15524823 TI - Spin separation in cyclotron motion. AB - Charged carriers with different spin states are spatially separated in a two dimensional hole gas. Because of strong spin-orbit interaction, holes at the Fermi energy in GaAs have different momenta for two possible spin states traveling in the same direction, and, correspondingly, different cyclotron orbits in a weak magnetic field. Two point contacts, acting as a monochromatic source of ballistic holes and a narrow detector arranged in the magnetic focusing geometry are demonstrated to work as a tunable spin filter. PMID- 15524824 TI - Magnetic-field-induced suppression of electronic conduction in a superlattice. AB - We use a magnetic field applied along the axis of a semiconductor superlattice (SL) as a controllable means of creating a one-dimensional band structure. We demonstrate that the current flow through the SL is strongly suppressed when the electron motion perpendicular to the SL axis is strongly confined by the quantizing magnetic field. By modeling this behavior using semiclassical and nonequilibrium Green's function methods, we show that the observed quenching arises from a qualitative change in electron dynamics caused by increasing quantum confinement. PMID- 15524825 TI - Photoemission from stepped W(110): initial or final state effect? AB - The electronic structure of the (110)-oriented terraces of stepped W(331) and W(551) is compared to the one of flat W(110) using angle-resolved photoemission. We identify a surface-localized state which develops perpendicular to the steps into a repeated band structure with the periodicity of the step superlattices. It is shown that a final-state diffraction process rather than an initial-state superlattice effect is the origin of the observed behavior and why it does not affect the entire band structure. PMID- 15524826 TI - Role of spin-orbit splitting and dynamical fluctuations in the Si(557)-Au surface. AB - Our ab initio calculations show that spin-orbit coupling is crucial to understand the electronic structure of the Si(557)-Au surface. The spin-orbit splitting produces the two one-dimensional bands observed in photoemission, which were previously attributed to spin-charge separation in a Luttinger liquid. This spin splitting might have relevance for future device applications. We also show that the apparent Peierls-like transition observed in this surface by scanning tunneling microscopy is a result of the dynamical fluctuations of the step-edge structure which are quenched as the temperature is decreased. PMID- 15524827 TI - Imaging of cyclotron emission from edge channels in quantum Hall conductors. AB - A local probing technique of cyclotron emission is applied to image nonequilibrium electrons generated along edge channels in quantum Hall conductors. In a lower-magnetic field region of a quantum Hall state plateau (filling factor 2 or approximately 5 TeV. PMID- 15524864 TI - Lattice study of the exotic s = +1 baryon. AB - We propose S = +1 baryon interpolating operators, which are based on an exotic description of the antidecuplet baryon, like the diquark-diquark-antiquark structure. By using one of the new operators, the mass spectrum of the spin-1/2 pentaquark states is calculated in quenched lattice QCD at beta = 6/g(2) = 6.2 on a 32(3) x 48 lattice. It is found that the J(P) assignment of the lowest Theta(uudds) state is most likely (1/2)(-). We also calculate the mass of the charm analog of the Theta and find that the Theta(c)(uuddc) state lies much higher than the DN threshold, in contrast to several model predictions. PMID- 15524865 TI - Infrared behavior and fixed points in Landau-gauge QCD. AB - We investigate the infrared behavior of gluon and ghost propagators in Landau gauge QCD by means of an exact renormalization group equation. We explain how, in general, the infrared momentum structure of Green functions can be extracted within this approach. An optimization procedure is devised to remove residual regulator dependences. In Landau-gauge QCD this framework is used to determine the infrared leading terms of the propagators. The results support the Kugo-Ojima confinement scenario. Possible extensions are discussed. PMID- 15524866 TI - Perturbative generation of a strange-quark asymmetry in the nucleon. AB - We point out that perturbative evolution in QCD at three loops generates a strange-antistrange asymmetry s(x) - s(x) in the nucleon's sea just from the fact that the nucleon has nonvanishing up and down quark valence densities. The recently computed three-loop splitting functions allow for an estimate of this effect. We find that a fairly sizable asymmetry may be generated. Results for analogous asymmetries in the heavy-quark sector are also presented. PMID- 15524869 TI - Physical significance of q deformation and many-body interactions in nuclei. AB - The quantum deformation concept is applied to a study of pairing correlations in nuclei with mass 40< or =A< or =100. While the nondeformed limit of the theory provides a reasonable overall description of certain nuclear properties and fine structure effects, the results show that the q deformation plays a significant role in understanding higher-order effects in the many-body interaction. PMID- 15524870 TI - Alternative interpretation of sharply rising E0 strengths in transitional regions. AB - It is shown that strong 0(+)(2)-->0(+)(1) E0 transitions provide a clear signature of phase transitional behavior in finite nuclei. Calculations using the interacting-boson approximation (IBA) show that these transition strengths exhibit a dramatic and robust increase in spherical-deformed shape transition regions, that this rise matches well the existing data, that the predictions of these E0 transitions remain large in deformed nuclei, that they arise from the specific d-boson coherence in the wave functions, and do not necessarily require the explicit mixing of normal and intruder configurations from different IBA spaces. PMID- 15524867 TI - Measurement of the generalized forward spin polarizabilities of the neutron. AB - The generalized forward spin polarizabilities gamma(0) and delta(LT) of the neutron have been extracted for the first time in a Q2 range from 0.1 to 0.9 GeV2. Since gamma(0) is sensitive to nucleon resonances and delta(LT) is insensitive to the Delta resonance, it is expected that the pair of forward spin polarizabilities should provide benchmark tests of the current understanding of the chiral dynamics of QCD. The new results on delta(LT) show significant disagreement with chiral perturbation theory calculations, while the data for gamma(0) at low Q2 are in good agreement with a next-to-leading-order relativistic baryon chiral perturbation theory calculation. The data show good agreement with the phenomenological MAID model. PMID- 15524871 TI - Angular dependence of the dipole-dipole interaction in a nearly one-dimensional sample of Rydberg atoms. AB - Atoms in an ultracold highly excited sample are strongly coupled through the dipole-dipole interaction. In an effort to understand and manipulate the complicated interactions in this system we are investigating their dependence on the relative orientation of the dipoles. By focusing a 480 nm beam from a tunable dye laser into a magneto-optical trap, we produce a nearly one-dimensional sample of Rydberg atoms. The trap lies at the center of four conducting rods with which we can vary the magnitude and direction of the electric field at the trap, thus controlling the orientation of the dipoles with respect to the sample axis. We have measured the strength of the interaction for a variety of relative orientations. PMID- 15524872 TI - Transitions driven by a missing frequency. AB - We demonstrate an intricate combination of strong and weak interactions of a broadband half-cycle pulse with Rydberg atoms. A transition, which is resonant with a frequency that was taken out of the frequency spectrum, can be enhanced. In the experiment, the broadband ultrashort terahertz half-cycle pulse propagates through water vapor, which absorbs on discrete lines in the spectrum, thus removing frequencies from the spectrum. The resulting pulse interacts with a rubidium Rydberg atom, which has a resonant transition with one of the missing frequencies. Under certain conditions, the transition probability is enhanced although the associated frequency was missing in the spectrum. PMID- 15524874 TI - Optimization of effective atom centered potentials for london dispersion forces in density functional theory. AB - We add an effective atom-centered nonlocal term to the exchange-correlation potential in order to cure the lack of London dispersion forces in standard density functional theory. Calibration of this long-range correction is performed using density functional perturbation theory and an arbitrary reference. Without any prior assignment of types and structures of molecular fragments, our corrected generalized gradient approximation density functional theory calculations yield correct equilibrium geometries and dissociation energies of argon-argon, benzene-benzene, graphite-graphite, and argon-benzene complexes. PMID- 15524873 TI - Kerr effect in liquid helium at temperatures below the superfluid transition. AB - The electro-optical Kerr effect induced by a slowly varying electric field in liquid helium at temperatures below the lambda point is investigated. The Kerr constant of liquid helium is measured to be (1.43+/-0.02(stat)+/-0.04(sys)) x 10( 20) (cm/V)(2) at T=1.5 K. Within experimental uncertainty, the Kerr constant is independent of temperature in the range T=1.5 K to 2.17 K, which implies that the Kerr constant of the superfluid component of liquid helium is the same as that of normal liquid helium. Pair and higher correlations of He atoms in the liquid phase account for about 23% of the measured Kerr constant. Liquid nitrogen was used to test the experimental setup; the result for the liquid nitrogen Kerr constant is (4.38+/-0.15) x 10(-18) (cm/V)(2). Kerr effect can be used as a noncontact technique for measuring the magnitude and mapping out the distribution of electric fields inside these cryogenic insulants. PMID- 15524875 TI - First observation of four-body breakup in electron recombination: C2D+5. AB - We report the first observation of four-body breakup in electron dissociative recombination of a molecular ion: C2D+5. In an ion storage ring experiment, the branching ratio for the process C2D+5 + e(-)-->C2D2 + D + D + D was determined to be 13%. This means that three covalent chemical bonds are broken as a result of the action of a single electron. This is the first time a four-body breakup of chemical bonds has been observed in a low-energy binary reaction. PMID- 15524876 TI - Three-body dissociation dynamics of the low-lying Rydberg states of H3 and D3. AB - The dynamics of the three-body dissociative charge exchange of fast (12 keV) H+3 and D+3 with Cs have been studied using multiparticle translational spectroscopy. The observed partitioning of product momenta was found to be state-specific and yields insights into the nuclear motion during dissociation for the three lowest lying 2s 2A'1, 2p 2A''2, and 3p 2E' metastable Rydberg states of H3 and the 2s 2A'1 and 2p 2A''2 states for D3. These results provide direct empirical information on the nonadiabatic couplings that govern the three-body dissociation of the lowest-lying Rydberg states of H3 and D3. PMID- 15524868 TI - Bose-Einstein correlations of charged pion pairs in Au + Au collisions at square root sNN = 200 GeV. AB - Bose-Einstein correlations of identically charged pion pairs were measured by the PHENIX experiment at midrapidity in Au + Au collisions at square root s(NN)=200 GeV. The Bertsch-Pratt radius parameters were determined as a function of the transverse momentum of the pair and as a function of the centrality of the collision. Using the standard core-halo partial Coulomb fits, and a new parametrization which constrains the Coulomb fraction as determined from the unlike-sign pion correlation, the ratio R(out)/R(side) is within 0.8-1.1 for 0.25< <1.2 GeV/c. The centrality dependence of all radii is well described by a linear scaling in N(1/3)(part), and R(out)/R(side) for approximately 0.45 GeV/c is approximately constant at unity as a function of centrality. PMID- 15524877 TI - High-efficiency detection of a single quantum of angular momentum by suppression of optical pumping. AB - We propose and demonstrate experimentally the discrimination between two spin states of an atom purely on the basis of their angular momentum. The discrimination relies on angular momentum selection rules and does not require magnetic effects such as a magnetic dipole moment of the atom or an applied magnetic field. The central ingredient is to prevent by coherent population trapping an optical pumping process which would otherwise relax the spin state before a detectable signal could be obtained. We detected the presence or absence of a single quantum (h) of angular momentum in a trapped calcium ion in a single observation with success probability 0.86. As a practical technique, the method can be applied to read out some types of quantum computer. PMID- 15524878 TI - Formation and annihilation of laser light pulse quanta in a thermodynamic-like pathway. AB - We present a theoretical and experimental study of multiple pulse formation in passively mode-locked lasers. Following a statistical-mechanics approach, the study yields a thermodynamic-like "phase diagram" with boundaries representing cascaded first order phase transitions. They correspond to abrupt creation or annihilation of pulses and a quantized rf power behavior, as system parameters (noise and/or pumping levels) are varied, in excellent accordance with the experiments. Remarkably, individual pulses carry an almost constant quantum of energy. PMID- 15524879 TI - Nonlinear unbalanced bessel beams: stationary conical waves supported by nonlinear losses. AB - Nonlinear losses accompanying self-focusing substantially impact the dynamic balance of diffraction and nonlinearity, permitting the existence of localized and stationary solutions of the 2D + 1 nonlinear Schrodinger equation, which are stable against radial collapse. These are featured by linear, conical tails that continually refill the nonlinear, central spot. An experiment shows that the discovered solution behaves as a strong attractor for the self-focusing dynamics in Kerr media. PMID- 15524880 TI - Tunable soliton self-bending in optical lattices with nonlocal nonlinearity. AB - We address the phenomenon of soliton self-bending in Kerr-type nonlocal nonlinear media with an imprinted transverse periodic modulation of the linear refractive index. We show that the imprinted optical lattice makes possible to control the mobility of soliton by varying the depth and the frequency of the linear refractive index modulation. PMID- 15524881 TI - Statistics of defect-mediated turbulence influenced by noise. AB - The influence of white noise on defect-mediated turbulence which is modeled by the complex Ginzburg-Landau equation is investigated. We show that the dynamics of defects in the noise-driven spatiotemporal chaos can be described by a simple statistical model. The noise enhances significantly the ability of the turbulent background to advocate new defects with a constant rate, and at the same time it increases the vanishing of defects in the system by introducing an additional annihilation rate that is proportional to the number of defects. A universal probability distribution function is derived for the number of defect pairs. PMID- 15524882 TI - Quantum-to-classical crossover of quasibound states in open quantum systems. AB - In the semiclassical limit of open ballistic quantum systems, we demonstrate the emergence of instantaneous decay modes guided by classical escape faster than the Ehrenfest time. The decay time of the associated quasibound states is smaller than the classical time of flight. The remaining long-lived quasibound states obey random-matrix statistics, renormalized in compliance with the recently proposed fractal Weyl law for open systems [W.T. Lu, S. Sridhar, and M. Zworski, Phys. Rev. Lett. 91, 154101 (2003)]. We validate our theory numerically for a model system, the open kicked rotator. PMID- 15524883 TI - Phase synchronization from noisy univariate signals. AB - We present methods for detecting phase synchronization of two unidirectionally coupled, self-sustained noisy oscillators from a signal of the driven oscillator alone. One method detects soft phase locking; another hard phase locking. Both are applied to the problem of detecting phase synchronization in von Karman vortex flow meters. PMID- 15524884 TI - Anomalous behavior of the coefficient of normal restitution in oblique impact. AB - The coefficient of normal restitution in an oblique impact is theoretically studied. Using a two-dimensional lattice model for an elastic disk and an elastic wall, we demonstrate that the coefficient of normal restitution can exceed unity and has a peak against the incident angle in our simulation. We also explain this behavior based upon a phenomenological theory. PMID- 15524885 TI - Two-dimensional locally resonant phononic crystals with binary structures. AB - The lumped-mass method is applied to study the propagation of elastic waves in two-dimensional binary periodic systems, i.e., periodic soft rubber/epoxy and vacuum/epoxy composites, for which the conventional methods fail or converge very slowly. A comprehensive study is performed for the two-dimensional binary locally resonant phononic crystals, which are composed of periodic soft rubber cylinders immersed in epoxy host. Numerical simulations predict that subfrequency gaps also appear because of the high contrast of mass density and elastic constant of the soft rubber. The locally resonant mechanism in forming the subfrequency gaps is thoroughly analyzed by studying the two-dimensional model and its quasi-one dimensional mechanical analog. The rule used to judge whether a resonant mode in the phononic crystals can result in a corresponding subfrequency gap or not is found. PMID- 15524886 TI - Codalike multiple scattering of elastic waves in dense granular media. AB - We study the multiple scattering of short-wavelength ultrasound through the force networks in dry and wet glass bead packings under stress. Over long distance scales, the diffusion approximation is shown to describe adequately the transport of elastic waves dominated by shear waves. The recovered transport mean path reveals a short-range correlation of the force chains. Also we observe the drastic effect of wetting liquids on the energy dissipation in the granular medium. The relevance of these experimental findings for the seismological applications is discussed. PMID- 15524887 TI - Eddy viscosity for time reversing waves in a dissipative environment. AB - We present new results for the time reversal of weakly nonlinear pulses traveling in a random dissipative environment. Also we describe a new theory for calculating the eddy viscosity for weakly nonlinear waves propagating over a random surface. The turbulent viscosity is calculated from first principles, namely, without imposing any stress-strain hypothesis. A viscous shallow water model is considered and its effective viscosity characterized. We also show that weakly nonlinear waves can still be time reversed under weak dissipation. Incoherently scattered signals are recompressed, both for time reversal in transmission as well as in reflection. Under the weakly nonlinear, weakly dissipative regime, dissipation only affects the refocused pulse profile regarding its amplitude, but its shape is not corrupted. Numerical experiments are presented. PMID- 15524888 TI - Crossover in the enstrophy decay in two-dimensional turbulence in a finite box. AB - The numerical simulation of two-dimensional decaying turbulence in a large but finite box presented in this Letter uncovered two physically different regimes of enstrophy decay. During the initial stage, the enstrophy Omega, generated by a random Gaussian initial condition, decays as Omega(t) proportional variant t( gamma) with gamma approximately 0.7-0.8. After that, the flow undergoes a transition to a gas or fluid composed of distinct vortices. Simultaneously, the magnitude of the decay exponent crosses over to gamma approximately 0.4. A theory predicting N(t) proportional variant t(-xi) and the magnitudes of exponents gamma=2/5 and xi=4/5 is presented. PMID- 15524889 TI - Velocity imaging of highly turbulent gas flow. AB - We introduce a noninvasive, quantitative magnetic resonance imaging (MRI) wind tunnel measurement in flowing gas (>10 m s(-1)) at high Reynolds numbers (Re>10(5)). The method pertains to liquids and gases, is inherently three dimensional, and extends the range of Re to which MRI is applicable by orders of magnitude. There is potential for clear time savings over traditional pointwise techniques. The mean velocity and turbulent diffusivity of gas flowing past a bluff obstruction and a wing section at realistic stall speeds were measured. The MRI data are compared with computational fluid dynamics. PMID- 15524890 TI - Spectroscopic diagnosis of nested-wire-array dynamics and interpenetration at 7 MA. AB - Nested-wire array experiments have been conducted at the 7 MA level with 150 ns implosion times from an outer diameter of 40 mm. Analysis of spectral data indicates that material from the outer array preferentially occupies the high temperature core of the stagnated pinch independent of the interwire gap in the range of 1.1 to 4.5 mm. PMID- 15524891 TI - Wave dispersion relation of two-dimensional plasma crystals in a magnetic field. AB - The wave dispersion relation in a two-dimensional strongly coupled plasma crystal is studied by theoretical analysis and molecular dynamics simulation taking into account a constant magnetic field parallel to the crystal normal. The expression for the wave dispersion relation clearly shows that high-frequency and low frequency branches exist as a result of the coupling of longitudinal and transverse modes due to the Lorenz force acting on the dust particles. The high frequency and the low-frequency branches are found to belong to right-hand and left-hand polarized waves, respectively. PMID- 15524892 TI - Ion acceleration by collisionless shocks in high-intensity-laser-underdense plasma interaction. AB - Ion acceleration by the interaction of an ultraintense short-pulse laser with an underdense-plasma has been studied at intensities up to 3 x 10(20) W/cm(2). Helium ions having a maximum energy of 13.2+/-1.0 MeV were measured at an angle of 100 degrees from the laser propagation direction. The maximum ion energy scaled with plasma density as n(0.70+/-0.05)(e). Two-dimensional particle-in-cell simulations suggest that multiple collisionless shocks are formed at high density. The interaction of shocks is responsible for the observed plateau structure in the ion spectrum and leads to an enhanced ion acceleration beyond that possible by the ponderomotive potential of the laser alone. PMID- 15524893 TI - Shear flows and shear viscosity in a two-dimensional Yukawa system (dusty plasma). AB - The shear viscosity of a two-dimensional liquid-state dusty plasma was measured experimentally. A monolayer of highly charged polymer microspheres, with a Yukawa interaction, was suspended in a plasma sheath. Two counterpropagating Ar+ laser beams pushed the particles, causing shear-induced melting of the monolayer and a shear flow in a planar Couette configuration. By fitting the particle velocity profiles in the shear flow to a Navier-Stokes model, the kinematic viscosity was calculated; it was of order 1 mm(2) s(-1), depending on the monolayer's parameters and shear stress applied. PMID- 15524894 TI - Spectral distribution of the cross helicity in the solar wind. AB - There are a variety of theoretical and observational indications that fluctuation energy in astrophysical and space plasma turbulence is distributed anisotropically in space relative to the magnetic field direction. The cross helicity, represented by correlations between velocity and magnetic field fluctuations, enters a magnetohydrodynamic description on equal footing with the energy, but its anisotropy has not been examined in the same degree of detail. Here we employ Advanced Coronal Explorer data to examine the rotational symmetry of the cross helicity. We find that the normalized cross helicity is associated more or less equally with all angular components of the fluctuations. This favors turbulence models that allow for cross communication between parallel and perpendicular wave numbers, suggesting that "wavelike" and "turbulencelike" fluctuations are strongly coupled. PMID- 15524895 TI - Spectral dips in ion emission emerging from ultrashort laser-driven plasmas. AB - Deep dips in MeV ion spectra are obtained from water droplet targets irradiated by intense [(0.5-1.2) x 10(19) W/cm(2)] and ultrashort (35 fs) laser pulses. The existence of these dips is ascribed to the generation of a multielectron temperature plasma, which is confirmed by our experiments. An existing fluid model based on hot-electron components with significantly different temperatures is consistent with the behavior we observe in the ion spectra of the femtosecond laser-driven interaction. The model provides a good simulation of the observed spectral dips and allows us to establish important parameters such as hot- and cold-electron temperatures and the respective electron density ratios. The result may be of interest for spectral tailoring of proton spectra in future applications of laser-generated proton beams. PMID- 15524896 TI - Theory of tokamak equilibria with central current density reversal. AB - It is found that, with a model current profile, the Grad-Shafranov equation can be reduced to the Helmholtz equation, which can describe a variety of equilibrium configurations. With the eigenvalue problem solved in the toroidal coordinate system, an analytical solution to the Grad-Shafranov equation is found. It is demonstrated that current reversal equilibrium configurations exist with finite radial gradient of plasma pressure and continuous current density, and that current density reversal is accompanied by pressure gradient reversal. PMID- 15524897 TI - Phase diagram of superfluid 3He in 99.3% porosity aerogel. AB - We report on continuous-wave NMR measurements of the energy gaps of the A-like and B-like superfluid phases of 3He at 28.4 mT confined to a 99.3% porosity silica aerogel. The gaps are suppressed by the presence of the aerogel in a temperature-independent manner, but the suppression is considerably stronger than expected from the suppression of T(c). We then use our measurements to calculate the free energy ratio between the A-like and B-like phases. The equilibrium AB transition temperature, derived from where this ratio reaches unity, is consistent with previous measurements of the initial displacement of the pinned AB interface on warming. On this basis, we present for the first time the equilibrium phase diagram of the A-like and B-like phases of superfluid 3He in aerogel. PMID- 15524898 TI - Casimir forces, surface fluctuations, and thinning of superfluid film. AB - Recent experiments on the wetting of 4He have shown that the film becomes thinner at the lambda transition and in the superfluid phase. The difference in thickness above and below the transition has been attributed to a Casimir interaction which is a consequence of a broken continuous symmetry in the bulk superfluid. However, the observed thinning of the film is larger than can be accounted by this Casimir force. We show that surface fluctuations give rise to an additional force, similar in form but larger in magnitude, which may explain the observations. PMID- 15524899 TI - Ring exchanges and the supersolid phase of 4He. AB - Using path integral Monte Carlo simulations we calculate exchange frequencies in bulk hcp 4He as atoms undergo ring exchange. We fit the frequencies to a lattice model and examine whether such atoms could become a supersolid, that is, have a nonclassical rotational inertia. We find that the scaling with respect to the number of exchanging atoms is such that superfluid behavior will not be observed in a perfect 4He crystal. PMID- 15524900 TI - Antiferromagnetic coupling driven by bond length contraction near the Ga1-xMnxN film surface. AB - Using first principles calculations based on gradient corrected density functional theory we show that Mn atoms, which couple ferromagnetically in bulk Ga1-xMnxN, couple antiferromagnetically on its surface. This change in magnetic behavior is brought about by a contraction of the Mn-Mn and Mn-N bond lengths, which is significantly greater on the surface than in the bulk. The present study provides new insight to the numerous conflicting experimental observations in Mn doped GaN systems. PMID- 15524901 TI - An interstitialcy theory of structural relaxation and related viscous flow of glasses. AB - A theory of isothermal structural relaxation and creep of glasses below the glass transition temperature is given. According to the interstitialcy theory, the supercooled liquid state does not exist below a Kauzmann "pseudocritical" temperature T(k), which lies above the temperature T(K), commonly called the "Kauzmann temperature." Structural relaxation is simply a reduction with time of the interstitialcy concentration to the crystalline state for TT(k). The predicted viscosity eta is universal, given by eta=eta(0) + eta(T)t, in agreement with experiment. eta is continuous in T, with eta discontinuous at T(k) but linear in 1/T above and below T(k). The dependence of eta on the shear modulus directly connects kinetic and thermodynamic properties of glasses and liquids. PMID- 15524902 TI - Lattice dynamics of an anharmonic crystal: evidence for interactions between atomic vibrations at high temperatures. AB - In studying the lattice dynamics of a strongly anharmonic crystal, we solve a set of nonlinear Langevin equations for interacting oscillators while a multiwell potential is calculated in the displacive limit from the first principles. The model applied to the peculiar vibrations of beta-Zr along [111] allows us to analyze all contributions to the spectral density and their influence on each other. We predict the effect of induced anharmonicity for quick vibrations due to their interaction with intrinsically anharmonic slow vibrations. This effect results in the broadband distribution in energy of inelastic neutron scattering known as the symmetry-forbidden phonon-branch splitting. PMID- 15524903 TI - Breakdown of the perturbative renormalization group at certain quantum critical points. AB - It is shown that the presence of multiple time scales at a quantum critical point can lead to a breakdown of the loop expansion for critical exponents, since coefficients in the expansion diverge. Consequently, results obtained from finite order perturbative renormalization-group treatments may not be an approximation in any sense to the true asymptotic critical behavior. This problem manifests itself as a nonrenormalizable field theory, or, equivalently, as the presence of a dangerous irrelevant variable. The quantum ferromagnetic transition in disordered metals provides an example. PMID- 15524904 TI - Finite-size effects on the structure of grain boundaries. AB - We present a combined experimental and theoretical analysis of the structure of finite-sized Sigma 3 [112] grain boundaries in Au. High-resolution electron microscopy shows lattice translations at the grain boundary, with the magnitude of the translation varying along the finite-sized grain boundaries. The presence of this structural profile is explained using continuum elasticity theory and first-principles calculations as originating from a competition between elastic energy and the energy cost of forming continuous [111] planes across the boundary. This competition leads to a structural transition between offset-free and nontrivial grain boundary structures at a critical grain boundary size, in agreement with the experiments. We also provide a method to estimate the energy barrier of the gamma surface. PMID- 15524905 TI - Role of nanostructured dual-oxide supports in enhanced catalytic activity: theory of CO oxidation over Au/IrO2/TiO2. AB - The synergetic effect in multicomponent catalysts is a topic of profound industrial importance and intense academic interest. On a newly identified multicomponent catalyst, Au/IrO(2)/TiO(2), first-principles density-functional theory is analyzed to clarify the outstanding catalytic activity of the system for oxidative reactions at high temperatures. By comparing CO oxidation on interfaces and single-component surfaces, it is revealed that a high dispersion of a more active oxide (IrO2), on a more inert oxide (TiO2) is the key. It preserves the sintering resistance of Au supported on less active oxides, while at the same time promoting oxidative reactions that occur at the Au/active-oxide interface. PMID- 15524906 TI - Observation of microscopic CO dynamics on Cu(001) using 3He spin-echo spectroscopy. AB - We present momentum resolved measurements of quasielastic helium atom scattering made using a new 3He spin-echo spectrometer. Our data for the dynamics of CO on Cu(001) indicates an activated jump mechanism which we analyze in detail using molecular dynamics simulations. A nearly isotropic potential energy surface is found with an average barrier height of approximately 125 meV, yielding comparable hopping rates along both the <110> and <100> directions. The measurements provide the first rigorous experimental test of state-of-the-art first-principles calculations previously made on this system. PMID- 15524907 TI - Chirality assignment of single-walled carbon nanotubes with strain. AB - Strain-induced band gap shifts that depend strongly on the chiral angle have been observed by optical spectroscopy in single-walled carbon nanotubes (SWCNTs). Uniaxial and torsional strains are generated by changing the environment surrounding the SWCNTs, using the surrounding D2O ice temperature or the hydration state of a wrapping polymer. These methods are used as diagnostic tools to determine the quantum number q and examine chiral vector indices for specific nanotubes. PMID- 15524908 TI - Spin reorientation and structural relaxation of atomic layers: pushing the limits of accuracy. AB - The correlation between an ad-layer-induced spin reorientation transition (SRT) and the ad-layer-induced structural relaxation is investigated by combined in situ surface x-ray diffraction and magneto-optical Kerr-effect experiments on Ni/Fe/Ni(111) layers on W(110). The Fe-induced SRT from in-plane to out-of-plane, and the SRT back to in-plane upon subsequent coverage by Ni, are each accompanied by a small lattice relaxation of at most 0.002 A. Such a small strain variation excludes a magnetoelasticity driven SRT, and we suggest the interface anisotropy as a possible driving force. PMID- 15524909 TI - Dilution-controlled quantum criticality in rare-earth nickelates. AB - A microscopic model for the diluted spin-mixed compounds (RxY1-x)2BaNiO5 (R=magnetic rare earth) is studied using quantum Monte Carlo simulations. The ordering temperature is shown to be a universal function of the impurity concentration x and the intrinsic Ni-chain correlation length. An effective model for the critical modes is derived. The possibility of a quantum critical point driven by the rare-earth concentration and the existence of a quantum Griffiths phase in the high dilution limit is investigated. Several possible experimental approaches to verify the results are put forward. PMID- 15524910 TI - Mutual experimental and theoretical validation of bulk photoemission spectra of Sr1-xCaxVO3. AB - We report high-resolution high-energy photoemission spectra together with parameter-free LDA + DMFT (local density approximation + dynamical mean-field theory) results for Sr1-xCaxVO3, a prototype 3d(1) system. In contrast to earlier investigations the bulk spectra are found to be insensitive to x. The good agreement between experiment and theory confirms the bulk sensitivity of the high energy photoemission spectra. PMID- 15524911 TI - Charge-orbital ordering and Verwey transition in magnetite. AB - Local density approximation + Hubbard U (LDA + U) band structure calculations reveal that magnetite (Fe3O4) forms an insulating charge-orbital-ordered state below the Verwey transition temperature. The calculated charge ordering is in good agreement with that inferred from recent experiments. We found an associated t(2g) orbital ordering on the octahedral Fe2+ sublattice. Such an orbital ordering results primarily from the on-site Coulomb interaction. This finding unravels such fundamental issues about the Verwey transition as the mechanism for the charge ordering and for the formation of the insulating gap, as well as the nonobedience of the Anderson's criterion for the charge ordering. PMID- 15524912 TI - Metal-dimer atomic reconstruction leading to deep donor states of the anion vacancy in II-VI and chalcopyrite semiconductors. AB - First-principles total-energy calculations reveal a novel local atomic reconstruction mode around anion vacancies in II-VI and chalcopyrite compounds resulting from the formation of metal dimers. As a consequence, the neutral Se vacancy has an unexpected low symmetry in ZnSe and becomes a deep donor in both ZnSe and CuGaSe2, contrary to the common belief regarding chalcopyrites. The calculated optical transition energies explain the hitherto puzzling absorption bands observed in the classic experiments of the color center in ZnS. PMID- 15524913 TI - Positron annihilation studies of the f-electron character in actinides. AB - Measurement of the angular correlation of the electron-positron annihilation radiation complemented with ab initio calculations can provide decisive information about the character of f electrons in actinide compounds. Our studies of the antiferromagnet UGa3 in the paramagnetic phase produce substantial evidence that an unconstrained f-electron itinerant description applies. PMID- 15524914 TI - Electronic structures of RTe2 (R = La,Ce): a clue to the pressure-induced superconductivity in CeTe1.82. AB - Electronic structures of RTe2 (R=La,Ce) have been investigated by using the local spin density approximation (LSDA) and the LSDA + U (U: on-site Coulomb interaction) band methods. Both LaTe2 and CeTe2 show the very similar Fermi surface nesting features along the [100] direction, which drive the charge density wave (CDW) instability in the Te(1) sheets. The contribution near E(F) from Ce 4f states is negligible in agreement with the measured ARPES spectra. In the semimetallic CDW-distorted RTe2, both Te vacancy and pressure induce the charge transfer from Te(1) 5p to R 5d states, producing the enhanced density of states at E(F). We suggest that these increased self-doped Te(1) 5p hole carriers are responsible for the pressure-induced superconductivity in nonstoichiometric CeTe1.82. PMID- 15524915 TI - Orbital and Spin Chains in ZnV2O4. AB - Our powder inelastic neutron scattering data indicate that ZnV2O4 is a system of spin chains that are three-dimensionally tangled in the cubic phase above 50 K due to randomly occupied t(2g) orbitals of V3+ (3d(2)) ions. Below 50 K in the tetragonal phase, the chains become straight due to antiferro-orbital ordering. This is evidenced by the characteristic wave vector dependence of the magnetic structure factor that changes from symmetric to asymmetric at the cubic-to tetragonal transition. PMID- 15524916 TI - Magnetite, a model system for mixed-valence oxides, does not show charge ordering. AB - We have investigated the charge ordering (CO) in magnetite below the Verwey transition. A new set of half-integer and mixed-integer superlattice reflections of the low-temperature phase have been studied by x-ray resonant scattering. None of these reflections show features characteristic of CO. We demonstrate the absence of CO along the c axis with the periodicity of either the cubic lattice q=(001) or the doubled cubic lattice q=(001/2). This result suggests that the Verwey transition is caused by strong electron-phonon interaction instead of an electronic ordering on the octahedral Fe atoms. PMID- 15524917 TI - Magnetic anisotropy of the antiferroquadrupole phase in Ce0.50La0.50B6. AB - Magnetic phase diagrams for antiferroquadrupole (AFQ) phase II and antiferromagnetic (AFM) phase III in Ce0.50La0.50B6 with a Gamma(8) ground state have been investigated by ultrasonic measurements. The hybrid magnet (Gama) in the National Institute for Materials Science was employed for high-field measurements up to 30 T and a 3He-4He dilution refrigerator was used for low temperature experiments down to 20 mK. The phase boundary from paramagnetic phase I to AFQ phase II under [001] magnetic fields closes at H(I-II) approximately 29 T, while the boundary is still open under fields along the [110] and [111] directions even up to 30 T. This anisotropic character of phase II in fields is consistent with the theoretical calculation based on the O(xy)-type AFQ ordering. We also found that AFM phase III reduces considerably in fields turning from the [001] to [110] and [111] directions. PMID- 15524918 TI - Role of surface States in scanning tunneling spectroscopy of (111) metal surfaces with Kondo adsorbates. AB - A nearly free-electron model to describe scanning tunneling spectroscopy of (111) metal surfaces with Kondo impurities is presented. Surface states are found to play an important role giving a larger contribution to the conductance of Cu(111) and Au(111) than Ag(111) surfaces. The different line shapes observed when Co is adsorbed on the different substrates are mainly determined by the position of the surface band onset relative to the Fermi energy and the decay length of the surface state into the substrate. The lateral dependence of the line shape amplitude is found to be bulklike for R|| < or approximately 3-5 A and surfacelike at larger distances, in agreement with experimental data. PMID- 15524919 TI - Role of strain and ligand effects in the modification of the electronic and chemical properties of bimetallic surfaces. AB - Periodic density functional calculations are used to illustrate how the combination of strain and ligand effects modify the electronic and surface chemical properties of Ni, Pd, and Pt monolayers supported on other transition metals. Strain and the ligand effects are shown to change the width of the surface d band, which subsequently moves up or down in energy to maintain a constant band filling. Chemical properties such as the dissociative adsorption energy of hydrogen are controlled by changes induced in the average energy of the d band by modification of the d-band width. PMID- 15524922 TI - Spin-Hall insulator. AB - Recent theories predict dissipationless spin current induced by an electric field in doped semiconductors. Nevertheless, the charge current is still dissipative in these systems. In this work, we theoretically predict the dissipationless spin Hall effect, without any accompanying charge current, in some classes of band insulators, including zero-gap semiconductors such as HgTe and narrow-gap semiconductors such as PbTe. This effect is similar to the quantum-Hall effect in that all the states below the gap contribute and there occurs no dissipation. However, the spin-Hall conductance is not quantized even in two dimensions. This is the first example of a nontrivial topological structure in a band insulator without any magnetic field. PMID- 15524921 TI - Time-resolved photoimaging of image-potential states in carbon nanotubes. AB - The first experimental evidence for the existence of image-potential states in carbon nanotubes is presented. The observed features constitute a new class of surface image states due to their quantized centrifugal motion. Measurements of binding energies and the temporal evolution of image state electrons were performed using femtosecond time-resolved photoemission. The associated lifetimes are found to be significantly longer than those of n=1 image state on graphite, indicating a substantial difference in electron decay dynamics between tubular and planar graphene sheets. PMID- 15524920 TI - Evidence of gap anisotropy in superconducting YNi2B2C using directional point contact spectroscopy. AB - We present a study of the anisotropy in the superconducting energy gap in a single crystal of YNi2B2C (T(c) approximately 14.6 K) using directional point contact spectroscopy. The superconducting energy gap at 2.7 K, when measured for I||c, is 4.5 times larger than that for I||a. The energy gaps in the two directions also have different temperature dependences. Our results support a scenario with s + g like symmetry. PMID- 15524923 TI - Two-flux composite fermion series of the fractional quantum Hall states in strained Si. AB - Magnetotransport properties are investigated in a high-mobility two-dimensional electron system in the strained Si quantum well of a (100) Si(0.75)Ge(0.25)/Si/Si(0.75)Ge0.25 heterostructure, at temperatures down to 30 mK and in magnetic fields up to 45 T. We observe around nu=1/2 the two-flux composite fermion (CF) series of the fractional quantum Hall effect (FQHE) at nu=2/3, 3/5, 4/7, and at nu=4/9, 2/5, 1/3. Among these FQHE states, the nu=1/3, 4/7, and 4/9 states are seen for the first time in the Si/SiGe system. Interestingly, of the CF series, the 3/5 state is weaker than the nearby 4/7 state and the 3/7 state is conspicuously missing, resembling the observation in the IQHE regime that the nu=3 is weaker than the nearby nu=4 state. Our results can be quantitatively understood in the picture of CF's with the valley degree of freedom. PMID- 15524924 TI - Anomalous proximity effect in gold coated (110) YBa2Cu3O7-delta films: penetration of the Andreev bound states. AB - Scanning tunneling spectroscopy of (110)YBa(2)Cu(3)O(7-delta)/Au bilayers reveal a proximity effect markedly different from the conventional one. While proximity induced mini-gaps rarely appear in the Au layer, the Andreev bound states clearly penetrate into the metal. Zero bias conductance peaks are measured on Au layers thinner than 7 nm with a magnitude similar to those detected on the bare superconductor films. The peaks then decay abruptly with Au thickness and disappear above 10 nm. This length is shorter than the normal coherence length and corresponds to the (ballistic) mean free path. PMID- 15524925 TI - Giant proximity effect in cuprate superconductors. AB - Using an advanced molecular beam epitaxy system, we have reproducibly synthesized atomically smooth films of high-temperature superconductors and uniform trilayer junctions with virtually perfect interfaces. We found that supercurrent runs through very thick barriers. We can rule out pinholes and microshorts; this "giant proximity effect" (GPE) is intrinsic. It defies the conventional explanation; it might originate in resonant tunneling through pair states in an almost-superconducting barrier. GPE may also be significant for superconducting electronics, since thick barriers are easier to fabricate. PMID- 15524926 TI - Crystal field potential of PrOs4Sb12: consequences for superconductivity. AB - The results of inelastic neutron scattering provide a solution for the crystal field level scheme in PrOs4Sb12, in which the ground state in the cubic crystal field potential of T(h) symmetry is a Gamma(1) singlet. The conduction electron mass enhancement is consistent with inelastic exchange scattering, and we propose that inelastic quadrupolar, or aspherical Coulomb, scattering is responsible for enhancing the superconducting transition temperature. PrOs4Sb12 appears to be the first compound in which aspherical Coulomb scattering is strong enough to overcome magnetic pair breaking and increase T(c). PMID- 15524927 TI - Pressure effects on the transition temperature and the magnetic field penetration depth in the pyrochlore superconductor RbOs2O6. AB - Magnetization measurements under hydrostatic pressure up to 8 kbar in the pyrochlore superconductor RbOs2O6 (T(c) approximately or equal 6.3 K at p=0) were carried out. A positive pressure effect on T(c) with dT(c)/dp=0.090(3) K/kbar was observed, whereas no pressure effect on the magnetic penetration depth lambda was detected. The pressure independent ratio 2 Delta(0)/k(B)T(c)=3.72(2) (Delta(0) is the superconducting gap at zero temperature) was found to be close to the BCS value 3.52. Magnetization and muon-spin rotation measurements of lambda(T) indicate that RbOs2O6 is an adiabatic s-wave BCS-type superconductor. The value of lambda extrapolated to zero temperature and ambient pressure was estimated to be 230(30) nm. PMID- 15524928 TI - NMR-like control of a quantum bit superconducting circuit. AB - Coherent superpositions of quantum states have already been demonstrated in different superconducting circuits based on Josephson junctions. These circuits are now considered for implementing quantum bits. We report on experiments in which the state of a qubit circuit, the quantronium, is efficiently manipulated using methods inspired from nuclear magnetic resonance (NMR): multipulse sequences are used to perform arbitrary operations, to improve their accuracy, and to fight decoherence. PMID- 15524929 TI - Magic angle effects and angular magnetoresistance oscillations as dimensional crossovers. AB - Interference effects between velocity and density of states, which occur as electrons move along open orbits in the extended Brillouin zone in anisotropic conductors, result in a change of wave functions' dimensionality at magic angle (MA) directions of a magnetic field. In particular, these 1D-->2D dimensional crossovers result in the appearance of sharp minima in a resistivity component rho perpendicular (H,alpha), perpendicular to conducting layers. This explains the main qualitative features of MA and angular magnetoresistance oscillations' phenomena observed due to the existence of quasi-one-dimensional sheets of Fermi surface in (TMTSF)2X, (DMET-TSeF)2X, and kappa-(ET)2Cu(NCS)(2) conductors. PMID- 15524930 TI - Oxygen superstructures throughout the phase diagram of (Y,Ca)Ba2Cu3O6+x. AB - The doping dependence of short-range lattice superstructures in (Y,Ca)Ba2Cu3O6+x has been studied with high-energy x-ray scattering. We observe diffuse features with a well defined periodicity which depend on the oxygen concentration but not on the charge carrier concentration. In addition, we find that diffuse scattering is absent in underdoped YBa2Cu4O8, which does not sustain oxygen defects. Our combined data highlight that the diffuse scattering arises from short-range oxygen ordering and associated lattice distortions. Signatures of stripe ordering or fluctuations are not seen and therefore must be much weaker. PMID- 15524931 TI - Four-unit-cell superstructure in the optimally doped YBa2Cu3O6.92 superconductor. AB - Diffuse x-ray scattering measurements reveal that the optimally doped YBa2Cu3O6.92 superconductor is intrinsically modulated due to the formation of a kinetically limited 4-unit-cell superlattice, q(0)=(1/4, 0, 0), along the shorter Cu-Cu bonds. The superlattice consists of large anisotropic displacements of Cu, Ba, and O atoms, respectively, which are correlated over approximately 3-6 unit cells in the ab plane, and appears to be consistent with the presence of an O ordered "ortho-IV" phase. Long-range strains emanating from these modulated regions generate an inhomogeneous lattice which may play a fundamentally important role in the electronic properties of yttrium-barium-copper-oxides. PMID- 15524932 TI - Nuclear ordering in lithium and an upper limit on its ambient pressure superconducting transition temperature. AB - We have discovered spontaneous ordering of nuclear spins in lithium metal by NMR measurements at very low temperatures. In low magnetic fields, B<0.2 mT, the NMR spectra show a pronounced low-frequency anomaly. Also, nonadiabatic response to a slowly varying magnetic field was observed. A rich phase diagram with three different nonparamagnetic regions is proposed. We estimate a critical spin temperature T(c) approximately 350 nK at B=0. We also report the absence of superconductivity in lithium at normal pressure down to T(e) approximately 100 microK (B<10 nT). PMID- 15524933 TI - Quantum superposition of high spin states in the single molecule magnet Ni4. AB - Quantum tunneling of the magnetization in a single molecule magnet has been studied in experiments that combine microwave spectroscopy with high sensitivity magnetic measurements. By monitoring spin-state populations in the presence of microwave radiation, the energy splittings between low lying superpositions of high-spin states of single molecule magnet Ni4 (S=4) have been measured. Absorption linewidths give an upper bound on the rate of decoherence. Pulsed microwave experiments provide a measure of energy relaxation time, which is found to increase with frequency. PMID- 15524934 TI - Controlled normal and inverse current-induced magnetization switching and magnetoresistance in magnetic nanopillars. AB - By combining pairs of ferromagnetic metals with the same or different signs of scattering anisotropies in ferromagnetic-nonmagnetic-ferromagnetic metal nanopillars, we independently invert just the magnetoresistance, just the direction of current-induced magnetization switching, or both together, at room temperature (295 K) and at 4.2 K. In all cases studied, the switching direction is correctly predicted from the net scattering anisotropy of the fixed ferromagnet, including both bulk and interfacial contributions. PMID- 15524935 TI - Finite-size effect on magnetic ordering temperatures in long-period antiferromagnets: holmium thin films. AB - The thickness dependence of the helical antiferromagnetic ordering temperature T(N) was studied for thin Ho metal films by resonant magnetic soft x-ray and neutron diffraction. In contrast with the Curie temperature of ferromagnets, T(N) was found to decrease with film thickness d according to [T(N)(infinity) T(N)(d)]/T(N)(d) proportional variant (d-d(0))(-lambda(')), where lambda(') is a phenomenological exponent and d(0) is of the order of the bulk magnetic period L(b). These observations are reproduced by mean-field calculations that suggest a linear relationship between d(0) and L(b) in long-period antiferromagnets. PMID- 15524936 TI - Some fermi surface properties of double-exchange interaction systems. AB - We study the photoemission spectrum of the double-exchange (DE) interaction systems. The DE Hamiltonian can be transformed into a simple form consisting of fermions and Schwinger bosons. We apply the gauge-field model and calculate the Green's function of the gauge field, fermions, and bosons. The imaginary part of the Green's function of an electron has an asymmetrical peak with strong temperature dependence. This can explain why the shape of the angle-resolved photoemission spectra of manganites near the Fermi surface is very different from that of Fermi liquid. We also show why the position of the Fermi surface is not sensitive to temperature. PMID- 15524937 TI - Structural, orbital, and magnetic order in vanadium spinels. AB - Vanadium spinels (ZnV2O4, MgV2O4, and CdV2O4) exhibit a sequence of structural and magnetic phase transitions, reflecting the interplay of lattice, orbital, and spin degrees of freedom. We offer a theoretical model taking into account the relativistic spin-orbit interaction, collective Jahn-Teller effect, and spin frustration. Below the structural transition, vanadium ions exhibit ferro-orbital order and the magnet is best viewed as two sets of antiferromagnetic chains with a single-ion Ising anisotropy. Magnetic order, parametrized by two Ising variables, appears at a tetracritical point. PMID- 15524938 TI - Generation of dark and bright spin wave envelope soliton trains through self modulational instability in magnetic films. AB - The generation of dark spin wave envelope soliton trains from a continuous wave input signal due to spontaneous modulational instability has been observed for the first time. The dark soliton trains were formed from high dispersion dipole exchange spin waves propagated in a thin yttrium iron garnet film with pinned surface spins at frequencies situated near the dipole gaps in the dipole-exchange spin wave spectrum. Dark and bright soliton trains were generated for one and the same film through placement of the input carrier frequency in regions of negative and positive dispersion, respectively. Two unreported effects in soliton dynamics, hysteresis and period doubling, were also observed. PMID- 15524939 TI - Laser-induced above-band-gap transparency in GaAs. AB - We report the observation of large (approximately 40%) laser-induced above-band gap transparency in GaAs at room temperature. The induced transparency is present only during the pulse width of the driving midinfrared laser pulses and its spectral shape is consistent with a laser-induced blueshift of the band edge. Our simulations based on the dynamic Franz-Keldysh effect reproduce the salient features of the experimental results, demonstrating, in particular, that the amount of the band edge shift is approximately given by the ponderomotive potential. PMID- 15524940 TI - Electron-electron interaction effects on the optical excitations of semiconducting single-walled carbon nanotubes. AB - We report correlated-electron calculations of optically excited states in ten semiconducting single-walled carbon nanotubes with a wide range of diameters. Optical excitation occurs to excitons whose binding energies decrease with increasing nanotube diameter, and are smaller than the binding energy of an isolated strand of poly-(paraphenylene vinylene). The ratio of the energy of the second optical exciton polarized along the nanotube axis to that of the lowest exciton is smaller than the value predicted within single-particle theory. The experimentally observed weak photoluminescence is an intrinsic feature of semiconducting nanotubes. PMID- 15524941 TI - Depth-controlled beta-NMR of 8Li in a thin silver film. AB - Depth-controlled beta-NMR can be used to probe the magnetic properties of thin films and interfaces on a nanometer length scale. A 30 keV beam of highly spin polarized 8Li+ ions was slowed down and implanted into a 50 nm film of Ag deposited on a SrTiO3 substrate. A novel high field beta-NMR spectrometer was used to observe two well resolved resonances which are attributed to Li occupying substitutional and octahedral interstitial sites in the Ag lattice. The temperature dependence of the Knight shifts and spin relaxation rates are consistent with the Korringa law for a simple metal, implying that the NMR of implanted 8Li reflects the spin suspectibility of bulk metallic silver. PMID- 15524942 TI - Equation of state of a dense columnar liquid crystal. AB - An accurate description of a columnar liquid crystal of hard disks at high packing fractions is presented using an improved free-volume theory. It is shown that the orientational entropy of the disks in the (one-dimensional) fluid direction leads to a different high-density scaling pressure compared to the prediction from traditional cell theory. Excellent quantitative agreement is found with recent Monte Carlo simulation results for various thermodynamic and structural properties. PMID- 15524943 TI - Synchrotron diffraction study of deformation mechanisms in mineralized tendon. AB - The high stiffness and toughness of biomineralized tissues are related to the material deformation mechanisms at different levels of organization, from trabeculae and osteons at the micrometer level to the mineralized collagen fibrils at the nanometer length scale. Quantitatively little is known about the sub-micrometer deformation mechanisms under applied load. Using a parallel-fibred mineralized tissue from the turkey leg tendon as a model for the mineralized collagen fibrils, we used in situ tensile testing with synchrotron x-ray diffraction to measure the average fibril deformation with applied external strain. Diffraction peak splitting occurred at large strains, implying an inhomogeneous elongation of collagen fibrils. Scanning electron microscopy measurements lead us to conclude that the inhomogeneous mineralization in mineralized tendon is at the origin of the high fracture strain. PMID- 15524944 TI - Strong coupling of nonlinear electronic and biological oscillators: reaching the "amplitude death" regime. AB - Interaction between an electronic and a biological circuit has been investigated for a pair of electrically connected nonlinear oscillators, with a spontaneously oscillating olivary neuron as the single-cell biological element. By varying the coupling strength between the oscillators, we observe a range of behaviors predicted by model calculations, including a reversible low-energy dissipation "amplitude death" where the oscillations in the coupled system cease entirely. PMID- 15524945 TI - Shape selection in chiral self-assembly. AB - Many biological and synthetic materials self-assemble into helical or twisted aggregates. The shape is determined by a complex interplay between elastic forces and the orientation and chirality of the constituent molecules. We study this interplay through Monte Carlo simulations, with an accelerated algorithm motivated by the growth of an aggregate out of solution. The simulations show that the curvature changes smoothly from cylindrical to saddlelike as the elastic moduli are varied. Remarkably, aggregates of either handedness form from molecules of a single handedness, depending on the molecular orientation. PMID- 15524946 TI - Fission of a multiphase membrane tube. AB - A common mechanism for intracellular transport is the use of controlled deformations of the membrane to create spherical or tubular buds. While the basic physical properties of homogeneous membranes are relatively well known, the effects of inhomogeneities within membranes are very much an active field of study. Membrane domains enriched in certain lipids, in particular, are attracting much attention, and in this Letter we investigate the effect of such domains on the shape and fate of membrane tubes. Recent experiments have demonstrated that forced lipid phase separation can trigger tube fission, and we demonstrate how this can be understood purely from the difference in elastic constants between the domains. Moreover, the proposed model predicts time scales for fission that agree well with experimental findings. PMID- 15524947 TI - Probing molecular free energy landscapes by periodic loading. AB - Single molecule pulling experiments provide information about interactions in biomolecules that cannot be obtained by any other method. However, the reconstruction of the molecule's free energy profile from the experimental data is still a challenge, in particular, for the unstable barrier regions. We propose a new method for obtaining the full profile by introducing a periodic ramp and using Jarzynski's relation for obtaining equilibrium quantities from nonequilibrium data. Our simulated experiments show that this method delivers significant more accurate data than previous methods, under the constraint of equal experimental effort. PMID- 15524948 TI - Quantized spiral tip motion in excitable systems with periodic heterogeneities. AB - Meandering spiral tips in homogeneous reaction-diffusion systems are characterized by two generically incommensurate radii and frequencies. Here, we create periodic perturbations in space to induce a transition to commensurate radii and frequencies that exhibit a devil's staircase. The plateaus of the staircase correspond to pinned or complex periodic orbits of the spiral tip. PMID- 15524950 TI - Comment on "Universal decoherence in solids". PMID- 15524949 TI - Stabilization of large generalized Lotka-Volterra foodwebs by evolutionary feedback. AB - Conventional ecological models show that complexity destabilizes foodwebs, suggesting that foodwebs should have neither large numbers of species nor a large number of interactions. However, in nature the opposite appears to be the case. Here we show that if the interactions between species are allowed to evolve within a generalized Lotka-Volterra model such stabilizing feedbacks and weak interactions emerge automatically. Moreover, we show that trophic levels also emerge spontaneously from the evolutionary approach, and the efficiency of the unperturbed ecosystem increases with time. The key to stability in large foodwebs appears to arise not from complexity per se but from evolution at the level of the ecosystem which favors stabilizing (negative) feedbacks. PMID- 15524951 TI - Comment on "Transient evolution of surface roughness on patterned GaAs(001) during homoepitaxial growth". PMID- 15524953 TI - Comment on "Anomalous conductance distribution in quasi-one-dimensional gold wires: possible violation of the one-parameter scaling hypothesis". PMID- 15524955 TI - Comment on "Instabilities of isotropic solutions of active polar filaments". PMID- 15524957 TI - Collective Rabi oscillations and solitons in a time-dependent BCS pairing problem. AB - Motivated by recent efforts to achieve cold fermions pairing, we study the nonadiabatic regime of the Bardeen-Cooper-Schrieffer state formation. After the interaction is turned on, at times shorter than the quasiparticle energy relaxation time, the system oscillates between the superfluid and normal state. The collective nonlinear evolution of the BCS-Bogoliubov amplitudes u(p), v(p), along with the pairing function Delta, is shown to be an integrable dynamical problem which admits single soliton and soliton train solitons. We interpret the collective oscillations as Bloch precession of Anderson pseudospins, where each soliton causes a pseudospin 2pi Rabi rotation. PMID- 15524958 TI - Reentrant phenomenon in the quantum phase transitions of a gas of bosons trapped in an optical lattice. AB - We calculate the location of the quantum phase transitions of a Bose gas trapped in an optical lattice as a function of effective scattering length a(eff) and temperature T. Knowledge of recent high-loop results on the shift of the critical temperature at weak couplings is used to locate a nose in the phase diagram above the free Bose-Einstein critical temperature T((0))(c), thus predicting the existence of a reentrant transition above T((0))(c), where a condensate should form when increasing a(eff). At zero temperature, the transition to the normal phase produces the experimentally observed Mott insulator. PMID- 15524959 TI - Dynamical role of anyonic excitation statistics in rapidly rotating bose gases. AB - We show that for rotating harmonically trapped Bose gases in a fractional quantum Hall state, the anyonic excitation statistics in the rotating gas can effectively play a dynamical role. For particular values of the two-dimensional coupling constant g=-2pih2(2k-1)/m, where k is a positive integer, the system becomes a noninteracting gas of anyons, with exactly obtainable solutions satisfying Bogomol'nyi self-dual order parameter equations. Attractive Bose gases under rapid rotation thus can be stabilized in the thermodynamic limit due to the anyonic statistics of their quasiparticle excitations. PMID- 15524960 TI - Current in open quantum systems. AB - We show that a dissipative current component is present in the dynamics generated by a Liouville-master equation, in addition to the usual component associated with Hamiltonian evolution. The dissipative component originates from coarse graining in time, implicit in a master equation, and needs to be included to preserve current continuity. We derive an explicit expression for the dissipative current in the context of the Markov approximation. Finally, we illustrate our approach with a simple numerical example, in which a quantum particle is coupled to a harmonic phonon bath and dissipation is described by the Pauli master equation. PMID- 15524961 TI - Matter-wave gap vortices in optical lattices. AB - We predict the existence of spatially localized nontrivial topological states of a Bose-Einstein condensate with repulsive atomic interactions confined by an optical lattice. These nonlinear localized states, matter-wave gap vortices, carry a vortexlike phase dislocation and exist in the gaps of the matter-wave band-gap spectrum due to the Bragg scattering. We discuss the structure, stability, and formation dynamics of the gap vortices in the case of two dimensional optical lattices. PMID- 15524962 TI - Dynamical instability of a doubly quantized vortex in a Bose-Einstein condensate. AB - Doubly quantized vortices were topologically imprinted in /F=1> 23Na condensates, and their time evolution was observed using a tomographic imaging technique. The decay into two singly quantized vortices was characterized and attributed to dynamical instability. The time scale of the splitting process was found to be longer at higher atom density. PMID- 15524963 TI - How to measure the decoherence of a micromaser field under well controlled conditions. AB - We discuss a possible realization of a quantum register with controllable decoherence in terms of /0> and /1> photon number states of a micromaser field. It is shown how to create in the Jaynes-Cummings model a superposition state of /0> and /1> photon number states inside a closed micromaser cavity. The loss of phase coherence between these two states can subsequently be measured by a second probe atom monitoring the decoherence of the field. A technique is proposed for forming the superposition of number states /0> and /1> using the time structure of the Rabi oscillation. The proposed method avoids problems with stray fields at the cavity holes, which disturb the coherence of the atomic superposition, and offers a way to study how the coupling strength to the environment influences the decoherence rate, displaying the robustness of physical qubits and the fidelity of quantum computations. PMID- 15524964 TI - Inconsistency in the application of the adiabatic theorem. AB - The adiabatic theorem states that an initial eigenstate of a slowly varying Hamiltonian remains close to an instantaneous eigenstate of the Hamiltonian at a later time. We show that a perfunctory application of this statement is problematic if the change in eigenstate is significant, regardless of how closely the evolution satisfies the requirements of the adiabatic theorem. We also introduce an example of a two-level system with an exactly solvable evolution to demonstrate the inapplicability of the adiabatic approximation for a particular slowly varying Hamiltonian. PMID- 15524965 TI - Morphological thermodynamics of fluids: shape dependence of free energies. AB - We examine the dependence of a thermodynamic potential of a fluid on the geometry of its container. If motion invariance, continuity, and additivity of the potential are satisfied, only four morphometric measures are needed to describe fully the influence of an arbitrarily shaped container on the fluid. These three constraints can be understood as a more precise definition for the conventional term extensive and have as a consequence that the surface tension and other thermodynamic quantities contain, aside from a constant term, only contributions linear in the mean and Gaussian curvature of the container and not an infinite number of curvatures as generally assumed before. We verify this numerically in the entropic system of hard spheres bounded by a curved wall. PMID- 15524966 TI - Suppression of noise in a noisy optical trap. AB - We have experimentally explored and clarified the concept of noise suppression in intrinsically noisy systems by adding noise at the input using a microscopic bead held in a moving and intensity-modulated optical trap. By coupling the stiffness of the optical trap to its position, we have explicitly constructed an experimental model system in which added fluctuations in the trap position result in reduced variance of the observed bead position as compared to a stationary trap. This reduction in variance and the spectral properties of the observed output noise agree with theoretical predictions. Our experiment demonstrates that the essential aspect of noise reduction in such a system is that the added fluctuations drive the system into states with a reduced intensity of intrinsic noise sufficiently often. PMID- 15524967 TI - Probing a nonequilibrium einstein relation in an aging colloidal glass. AB - We present a direct experimental measurement of an effective temperature in a colloidal glass of laponite, using a micrometric bead as a thermometer. The nonequilibrium fluctuation-dissipation relation, in the particular form of a modified Einstein relation, is investigated with diffusion and mobility measurements of the bead embedded in the glass. We observe an unusual nonmonotonic behavior of the effective temperature: starting from the bath temperature, it is found to increase up to a maximum value, and then decrease back, as the system ages. We show that the observed deviation from the Einstein relation is related to the relaxation times previously measured in dynamic light scattering experiments. PMID- 15524968 TI - Hyperpolarized xenon nuclear spins detected by optical atomic magnetometry. AB - We report the use of an atomic magnetometer based on nonlinear magneto-optical rotation with frequency-modulated light to detect nuclear magnetization of xenon gas. The magnetization of a spin-exchange-polarized xenon sample (1.7 c m(3) at a pressure of 5 bars, natural isotopic abundance, polarization 1% ), prepared remotely to the detection apparatus, is measured with an atomic sensor. An average magnetic field of approximately 10 nG induced by the xenon sample on the 10 cm diameter atomic sensor is detected with signal-to-noise ratio approximately 10 , limited by residual noise in the magnetic environment. The possibility of using modern atomic magnetometers as detectors of nuclear magnetic resonance and in magnetic resonance imaging is discussed. Atomic magnetometers appear to be ideally suited for emerging low-field and remote-detection magnetic resonance applications. PMID- 15524969 TI - Push-pull optical pumping of pure superposition states. AB - A new optical pumping method, "push-pull pumping," can produce very nearly pure, coherent superposition states between the initial and the final sublevels of the important field-independent 0-0 clock resonance of alkali-metal atoms. The key requirement for push-pull pumping is the use of D1 resonant light which alternates between left and right circular polarization at the Bohr frequency of the state. The new pumping method works for a wide range of conditions, including atomic beams with almost no collisions, and atoms in buffer gases with pressures of many atmospheres. PMID- 15524970 TI - Numerical simulations of generic singularities. AB - Numerical simulations of the approach to the singularity in vacuum spacetimes are presented here. The spacetimes examined have no symmetries and can be regarded as representing the general behavior of singularities. It is found that the singularity is spacelike and that, as it is approached, the spacetime dynamics becomes local and oscillatory. PMID- 15524971 TI - Evidence for nucleosynthesis in the supernova gamma process: universal scaling for p nuclei. AB - Analyzing the solar system abundance, we find two universal scaling laws concerning the p and s nuclei. They indicate that the gamma process in supernova (SN) explosions is the most probable origin of the p nuclei that has been discussed with many possible nuclear reactions and sites in about 50 years. In addition, the scalings lead to new concepts: a universality of the gamma process and a new nuclear cosmochronometer. We carry out gamma-process nucleosynthesis calculations for typical core-collapse SN explosion models, and the results satisfy the observed scalings. PMID- 15524973 TI - Mass measurement on the rp-process waiting point 72Kr. AB - The mass of one of the three major waiting points in the astrophysical rp process 72Kr was measured for the first time with the Penning trap mass spectrometer ISOLTRAP. The measurement yielded a relative mass uncertainty of deltam/m=1.2x10( 7) (deltam=8 keV). (73,74)Kr, also needed for astrophysical calculations, were measured with more than 1 order of magnitude improved accuracy. We use the ISOLTRAP masses of 72-74Kr to reanalyze the role of 72Kr (T(1/2)=17.2 s) in the rp process during x-ray bursts and conclude that 72Kr is a strong waiting point delaying the burst duration with at least 80% of its beta-decay half-life. PMID- 15524974 TI - Squeezing in the audio gravitational-wave detection band. AB - We demonstrate the generation of broadband continuous-wave optical squeezing from 280 Hz-100 kHz using a below-threshold optical parametric oscillator (OPO). The squeezed state phase was controlled using a noise locking technique. We show that low frequency noise sources, such as seed noise, pump noise, and detuning fluctuations, present in optical parametric amplifiers, have negligible effect on squeezing produced by a below-threshold OPO. This low frequency squeezing is ideal for improving the sensitivity of audio frequency measuring devices such as gravitational-wave detectors. PMID- 15524972 TI - Neutron capture cross section measurement of 151Sm at the CERN neutron time of flight facility (n_TOF). AB - The151Sm(n,gamma)152Sm cross section has been measured at the spallation neutron facility n_TOF at CERN in the energy range from 1 eV to 1 MeV. The new facility combines excellent resolution in neutron time-of-flight, low repetition rates, and an unsurpassed instantaneous luminosity, resulting in rather favorable signal/background ratios. The 151Sm cross section is of importance for characterizing neutron capture nucleosynthesis in asymptotic giant branch stars. At a thermal energy of kT=30 keV the Maxwellian averaged cross section of this unstable isotope (t(1/2)=93 yr) was determined to be 3100+/-160 mb, significantly larger than theoretical predictions. PMID- 15524975 TI - Model-independent assessment of current direct searches for spin-dependent dark matter. AB - I evaluate the current results of spin-dependent weakly interacting massive particle searches within a model-independent framework, showing the most restrictive limits to date derive from the combination of xenon and sodium iodide experiments. The extension of this analysis to the case of positive signal experiments is elaborated. PMID- 15524976 TI - Possible evidence for MeV dark matter in dwarf spheroidals. AB - The observed 511 keV emission from the galactic bulge could be due to very light (MeV) annihilating dark matter particles. To distinguish this hypothesis from conventional astrophysical sources, we study dwarf spheroidals in the region observed by the International Gamma-Ray Astrophysics Laboratory/SPI such as Sagittarius. As these galaxies have comparatively few stars, the prospects for 511 keV emission from standard astrophysical scenarios are minimal. The dwarf spheroidals do, however, contain copious amounts of dark matter. The observation of 511 keV emission from Sagittarius should be a "smoking gun" for MeV dark matter. PMID- 15524977 TI - Construction of non-Abelian walls and their complete moduli space. AB - We present a systematic method to construct exactly all Bogomol'nyi-Prasad Sommerfield multiwall solutions in supersymmetric U(N(C)) gauge theories in five dimensions with N(F) hypermultiplets in the fundamental representation for infinite gauge coupling. The moduli space of these non-Abelian walls is found to be the complex Grassmann manifold SU(N(F)) divided by SU(N(C))xSU(N(F)-N(C))xU(1) endowed with a deformed metric. PMID- 15524978 TI - Spin chains and string theory. AB - Recently, an important test of the anti de Sitter/conformal field theory correspondence has been done using rotating strings with two angular momenta. We show that such a test can be described more generally as the agreement between two actions: one a low energy description of a spin chain appearing in the field theory side, and the other a limit of the string action in AdS5xS5. This gives a map between the mean value of the spin in the boundary theory and the position of the string in the bulk, and shows how a string action can emerge from a gauge theory in the large-N limit. PMID- 15524979 TI - Relation between the neutrino and quark mixing angles and grand unification. AB - We argue that there exists a simple relation between the quark and lepton mixings, which supports the idea of grand unification and probes the underlying robust bimaximal fermion mixing structure of still unknown flavor physics. In this framework the quark mixing matrix is a parameter matrix describing the deviation of neutrino mixing from exactly bimaximal, predicting theta(sol)+theta(C)=pi/4, where theta(C) is the Cabibbo angle, theta(atm)+theta(CKM)(23)=pi/4 and theta(MNS)(13) approximately theta(CKM)(13) approximately O(lambda(3)), in perfect agreement with experimental data. Both non Abelian and Abelian flavor symmetries are needed for such a prediction to be realistic. An example flavor model capable of explaining this flavor mixing pattern and inducing the measured quark and lepton masses is outlined. PMID- 15524980 TI - Production of the X( 3872) in B-meson decay by the coalescence of charm mesons. AB - If the recently discovered charmonium state X( 3872) is a loosely bound S-wave molecule of the charm mesons D0 D(*0) or D(*0) D0, it can be produced in B-meson decay by the coalescence of charm mesons. If this coalescence mechanism dominates, the ratio of the differential rate for B+ -->D(0) D(* 0)K+ near the D0 D(*0) threshold and the rate for B+ -->XK+ is a function of the D0 D(*0) invariant mass and hadron masses only. The identification of the X( 3872) as a D0 D(*0)/D(*0)D0 molecule can be confirmed by observing an enhancement in the D0 D(*0) invariant mass distribution near the threshold. An estimate of the branching fraction for B+ -->XK+ is consistent with observations if X has quantum numbers J(PC)=1(++ ) and if J/psi pi(+) pi(-) is one of its major decay modes. PMID- 15524982 TI - Photon interferometry of Au+Au collisions at the BNL Relativistic Heavy-Ion Collider. AB - We calculate the two-body correlation function of direct photons produced in central Au+Au collisions at the Relativistic Heavy-Ion Collider. Our calculation includes contributions from the early preequilibrium phase in which photons are produced via hard parton scatterings as well as radiation of photons from a thermalized quark-gluon plasma and the subsequent expanding hadron gas. We find that high energy photon interferometry provides a faithful probe of the details of the space-time evolution and of the early reaction stages of the system. PMID- 15524983 TI - Evidence of complete fusion in the sub-barrier 16O+238U reaction. AB - Evaporation residue cross sections in the 16O+238U reaction were measured for the energy range from above- to extreme sub-barrier. The cross sections are reproduced by a statistical model calculation, for which partial cross sections are calculated by a coupled-channel model taking into account the prolate deformation of 238U. Complete fusion was observed in the collision of the projectile with the tips of the 238U target, in the same way as the side collision. PMID- 15524984 TI - Suppression of excitation and spectral broadening induced by interactions in a cold gas of Rydberg atoms. AB - We report on the observation of ultralong range interactions in a gas of cold rubidium Rydberg atoms. The van der Waals interaction between a pair of Rydberg atoms separated as far as 100,000 Bohr radii features two important effects: spectral broadening of the resonance lines and suppression of excitation with increasing density. The density dependence of these effects is investigated in detail for the S- and P-Rydberg states with principal quantum numbers n approximately 60 and n approximately 80 excited by narrow-band continuous-wave laser light. The density-dependent suppression of excitation can be interpreted as the onset of an interaction-induced local blockade. PMID- 15524985 TI - Conjugation length dependence of internal conversion in carotenoids: role of the intermediate state. AB - We report on a sub-20-fs transient absorption study of the S2(1(1)B(+)(u))- >S1(2(1)A(-)(g)) internal conversion in a series of carotenoids with a number of conjugated double bonds (N) ranging from 5 to 15. For the longer carotenoids (N>or=9), the measurements reveal the existence of an additional intermediate excited state lying between the optically allowed S2 state and the lower-lying forbidden S1 state. This state enables us to explain the nonmonotonic dependence of the S2-->S1 conversion rate on N and is expected to play an important role in photosynthetic light harvesting. PMID- 15524986 TI - Experimental observation of interatomic coulombic decay in neon dimers. AB - Recently Cederbaum et al. [Phys. Rev. Lett. 79, 4778 (1997)]] predicted a new decay channel of excited atoms and molecules termed interatomic Coulombic decay (ICD). In ICD the deexcitation energy is transferred via virtual photon exchange to a neighboring atom, which releases it by electron emission. We report on an experimental observation of ICD in 2s ionized neon dimers. The process is unambiguously identified by detecting the energy of two Ne1+ fragments and the ICD electron in coincidence, yielding a clean, background free experimental spectral distribution of the ICD electrons. PMID- 15524987 TI - "Eclipse" effect in the scattering of weakly bound helium clusters. AB - The total cross sections of the helium dimer, trimer, and tetramer for scattering from Kr atoms have been measured for cluster beam velocities between 250 and 820 m/s. The dimer cross section is twice that of the atom within 5% indicating that the Kr atoms scatter from the He atoms independently, which is consistent with the large dimer bond distance of about 50 A. The trimer and tetramer cross sections are somewhat larger and can be described by an impulse approximation with a multiple "eclipse" correction, extending ideas of Glauber for high energy collisions with the deuteron. PMID- 15524988 TI - Controlling HD+ and H+2 dissociation with the carrier-envelope phase difference of an intense ultrashort laser pulse. AB - Carrier-envelope phase difference effects in the dissociation of the HD+ molecular ion in the field of an intense, linearly polarized, ultrashort laser pulse are studied in the framework of the time-dependent Schrodinger equation. We consider a reduced-dimensionality model in which the nuclei are free to vibrate along the field polarization and the electrons move in two dimensions. The laser has a central wavelength of 790 nm and a pulse length of 10 fs with intensities in the range 6x10(14) to 9x10(14) W/cm(2). We find that the angular distribution of dissociation to p+D and H+d can be controlled by varying the phase difference, generating differences between the dissociation channels of more than a factor of 2. Moreover, the asymmetry is nearly as large for H+2 dissociation. PMID- 15524989 TI - Continuous weak measurement and nonlinear dynamics in a cold spin ensemble. AB - A weak continuous quantum measurement of an atomic spin ensemble can be implemented via Faraday rotation of an off-resonance probe beam, and may be used to create and probe nonclassical spin states and dynamics. We show that the probe light shift leads to nonlinearity in the spin dynamics and limits the useful Faraday measurement window. Removing the nonlinearity allows a nonperturbing measurement on the much longer time scale set by decoherence. The nonlinear spin Hamiltonian is of interest for studies of quantum chaos and real-time quantum state estimation. PMID- 15524990 TI - Optimization of attosecond pulse generation. AB - The generation of attosecond pulses by superposition of high harmonics relies on their synchronization in the emission. Our experiments in the low-order, plateau, and cutoff regions of the spectrum reveal different regimes in the electron dynamics determining the synchronization quality. The shortest pulses are obtained by combining a spectral filtering of harmonics from the end of the plateau and the cutoff, and a far-field spatial filtering that selects a single electron quantum path contribution to the emission. This method applies to isolated pulses as well as pulse trains. PMID- 15524991 TI - Modulational instability and parametric amplification induced by loss dispersion in optical fibers. AB - We show that modulational instability may arise even in the normal group-velocity dispersion regime of an optical fiber when the fiber loss (gain) varies depending on the wavelength. A simple analytical expression for the instability gain is obtained, which reveals that the odd-order terms of the loss dispersion are responsible for this phenomenon. The instability gain is measured experimentally in an optical-parametric-amplification configuration. Large parametric gain is induced in a non-phase-matched regime as we apply narrow band loss at the idler wavelength. PMID- 15524992 TI - Gravity-sensitive quantum dynamics in cold atoms. AB - We subject a falling cloud of cold cesium atoms to periodic kicks from a sinusoidal potential created by a vertical standing wave of laser light. By controllably accelerating the potential, we show quantum accelerator mode dynamics to be highly sensitive to the effective gravitational acceleration when close to specific, resonant values. This quantum sensitivity to a control parameter is reminiscent of that associated with classical chaos and promises techniques for precision measurement. PMID- 15524993 TI - Quasiscarred resonances in a spiral-shaped microcavity. AB - We study resonance patterns of a spiral-shaped dielectric microcavity with chaotic ray dynamics. Many resonance patterns of this microcavity, with refractive indices n=2 and 3, exhibit strong localization of simple geometric shape, and we call them quasiscarred resonances in the sense that there is, unlike conventional scarring, no underlying periodic orbits. It is shown that the formation of a quasiscarred pattern can be understood in terms of ray dynamical probability distributions and wave properties like uncertainty and interference. PMID- 15524994 TI - Multistability and memory effect in a highly turbulent flow: experimental evidence for a global bifurcation. AB - We report experimental evidence of a global bifurcation on a highly turbulent von Karman flow. The mean flow presents multiple solutions: the canonical symmetric solution becomes marginally unstable towards a flow which breaks the basic symmetry of the driving apparatus even at very large Reynolds numbers. The global bifurcation between these states is highly subcritical and the system thus keeps a memory of its history. The transition recalls low-dimension dynamical system transitions and exhibits very peculiar statistics. We discuss the role of turbulence in two ways: the multiplicity of hydrodynamical solutions and the effect of fluctuations on the nature of transitions. PMID- 15524995 TI - Monitoring Alfven cascades with interferometry on the JET Tokamak. AB - A microwave interferometry technique is applied for the first time for detecting a discrete spectrum of Alfven cascade (AC) eigenmodes excited with fast ions in reversed magnetic shear plasmas of the Joint European Torus. The interferometry measurements of plasma density perturbations associated with ACs show an unprecedented frequency and time resolution superior to that obtained with external magnetic coils. The measurements of ACs are used for monitoring the evolution of the safety factor and density of rational magnetic surfaces in the region of maximum plasma current. PMID- 15524996 TI - Identification of zonal flows in a toroidal plasma. AB - This Letter presents experimental confirmation of the presence of zonal flows in magnetically confined toroidal plasma using an advanced diagnostic system--dual heavy ion beam probes. The simultaneous observation of an electric field at two distant toroidal locations (approximately 1.5 m apart) in the high temperature (approximately 1 keV) plasma provides a fluctuation spectrum of electric field (or flow), a spatiotemporal structure of the zonal flows (characteristic radial length of approximately 1.5 cm and lifetime of approximately 1.5 ms), their long range correlation with toroidal symmetry (n=0), and the difference in the zonal flow amplitude with and without a transport barrier. These constitute essential elements of turbulence-zonal flow systems, and illustrate one of the fundamental processes of structure formation in nature. PMID- 15524997 TI - Effects of a vertical magnetic field on particle confinement in a magnetized plasma torus. AB - The particle confinement in a magnetized plasma torus with superimposed vertical magnetic field is modeled and measured experimentally. The formation of an equilibrium characterized by a parallel plasma current canceling out the grad B and curvature drifts is described using a two-fluid model. Characteristic response frequencies and relaxation rates are calculated. The predictions for the particle confinement time as a function of the vertical magnetic field are verified in a systematic experimental study on the TORPEX device, including the existence of an optimal vertical field and the anticorrelation between confinement time and density. PMID- 15524998 TI - Dust Coulomb balls: three-dimensional plasma crystals. AB - First experimental investigations of spherical three-dimensional plasma crystals consisting of hundreds or thousands of micrometer-sized polymer particles suspended in a radio-frequency gas discharge are described. These "Coulomb balls" are not subject to the formation of dust-free regions (voids) and have an unusual structure of nested crystalline shells. While small systems are in a solid phase, large systems show melting effects. PMID- 15524999 TI - Growth of a third ferromagnetic solid 3He layer on graphite. AB - We have measured the nuclear susceptibility of 3He in Grafoil filled with pure liquid 3He over the pressure region between 0.6 and 31.38 bars and at temperatures down to 0.5 mK with a cw NMR method. The nuclear magnetization corresponding to the adsorbed 3He layers on the Grafoil surface shows a strong ferromagnetic tendency with a periodic behavior as a function of liquid pressure. This observation is attributable to the growth of third and fourth solid 3He layers with the liquid pressure increase. The pressure dependence of the Weiss temperature indicates the third layer is completed at 19 bars and the fourth probably at 28 bars. The number of localized spins estimated from the solid magnetization is almost doubled from 0 to 28 bars, being consistent with this scenario. PMID- 15525000 TI - Bayesian ensemble approach to error estimation of interatomic potentials. AB - Using a Bayesian approach a general method is developed to assess error bars on predictions made by models fitted to data. The error bars are estimated from fluctuations in ensembles of models sampling the model-parameter space with a probability density set by the minimum cost. The method is applied to the development of interatomic potentials for molybdenum using various potential forms and databases based on atomic forces. The calculated error bars on elastic constants, gamma-surface energies, structural energies, and dislocation properties are shown to provide realistic estimates of the actual errors for the potentials. PMID- 15525001 TI - Structure of the fivefold surface of the icosahedral Al-Cu-Fe quasicrystal: experimental evidence of bulk truncations at larger interlayer spacings. AB - Based on scanning tunneling microscopy of the fivefold surface of the icosahedral Al-Cu-Fe quasicrystal and the refined structure model of the isostructural i-Al Pd-Mn, we present evidence that the surface corresponds to bulk truncations at the positions where blocks of atomic layers are separated by larger interlayer spacings (gaps). Both step-height distribution and high resolution scanning tunneling microscopy images on terraces reveal bulk truncations at larger gaps. PMID- 15525002 TI - Nanovoid cavitation by dislocation emission in aluminum. AB - This Letter is concerned with the determination of the transition paths attendant to nanovoid growth in aluminum under hydrostatic tension. The analysis is, therefore, based on energy minimization at 0 K. Aluminum is modeled by the Ercolessi-Adams embedded-atom method, and spurious boundary artifacts are mitigated by the use of the quasicontinuum method. Our analysis reveals several stages of pressure buildup separated by yield points. The first yield point corresponds to the formation of highly stable tetrahedral dislocation junctions around the surfaces of the void. The second yield point is caused by the dissolution of the tetrahedral structures and the emission of conventional 1/2<110>[111] and anomalous 1/2<110>[001] dislocation loops. PMID- 15524981 TI - Observation and properties of the X(3872) decaying to J/psipi(+)pi(-) in pp collisions at sqrt[s]=1.96 TeV. AB - We report the observation of the X(3872) in the J/psipi(+)pi(-) channel, with J/psi decaying to mu(+)mu(-), in pp collisions at sqrt[s]=1.96 TeV. Using approximately 230 pb(-1) of data collected with the Run II D0 detector, we observe 522+/-100 X(3872) candidates. The mass difference between the X(3872) state and the J/psi is measured to be 774.9+/-3.1(stat)+/-3.0(syst) MeV/c(2). We have investigated the production and decay characteristics of the X(3872) and find them to be similar to those of the psi(2S) state. PMID- 15525003 TI - Fusion of the extended modified liquid drop model for nucleation and dynamical nucleation theory. AB - We present a new phenomenological approach to nucleation, based on the combination of the "extended modified liquid drop" model and dynamical nucleation theory. The new model proposes a new cluster definition, which properly includes the effect of fluctuations, and it is consistent both thermodynamically and kinetically. The model is able to predict successfully the free energy of formation of the critical nucleus, using only macroscopic thermodynamic properties. It also accounts for the spinodal and provides excellent agreement with the result of recent simulations. PMID- 15525004 TI - Effective temperatures in driven systems: static versus time-dependent relations. AB - Using simulations of glassy systems under steady-state shear, we compare effective temperatures obtained from static linear response with those from time dependent fluctuation-dissipation relations. Although these two definitions are not expected to agree, we show that they yield the same answer over two and a half decades of effective temperature. This suggests that a more complete conceptual framework is necessary for effective temperatures in steady-state driven systems. PMID- 15525005 TI - Kinetic step pairing. AB - We report on the theoretical and experimental discovery of pairing of identical crystal steps. We first show that step bunching always occurs at long wavelength in the vicinity of an instability threshold when step dynamics is local. But an instability towards a stable train of pairs can be obtained when steps dynamics is nonlocal. This instability is shown to occur for transparent steps under electromigration. Observations on Si(111) under electromigration around 1230 degrees C show stable trains of pairs. By controlling both supersaturation and electromigration, we establish an experimental morphology diagram, from which we conclude that the transparency kinetic coefficient is negative. PMID- 15525006 TI - Enhanced self-diffusion on Cu(111) by trace amounts of s: chemical-reaction limited kinetics. AB - We find that less than 0.01 monolayer of S can enhance surface self-diffusion on Cu(111) by several orders of magnitude. The measured dependence of two dimensional island decay rates on S coverage (theta(S)) is consistent with the proposal that Cu3S3 clusters are responsible for the enhancement. Unexpectedly, the decay and ripening are diffusion limited with very low and very high theta(S) but not for intermediate theta(S). To explain this result we propose that surface mass transport in the intermediate region is limited by the rate of reaction to form Cu3S3 clusters on the terraces. PMID- 15525007 TI - Etchant anisotropy controls the step bunching instability in KOH etching of silicon. AB - STM investigations of vicinal Si(111) surfaces etched in KOH solutions under controlled flow conditions show that step bunching instability is due to inhomogeneities that develop in the etchant as the result of highly step-site specific etching reactions. Other previously postulated mechanisms for step bunching, including anisotropic surface diffusion, surface strain, and impurity deposition, are conclusively ruled out. The inhomogeneities locally accelerate etching near surface steps. Kinetic Monte Carlo simulations of this process qualitatively reproduce the observed morphologies. PMID- 15525008 TI - Systematic study of the effect of disorder on nanotribology of self-assembled monolayers. AB - The adhesion and friction between pairs of ordered and disordered self-assembled monolayers on SiO2 are studied using molecular dynamics. The disorder is introduced by randomly removing chains from a well ordered crystalline substrate and by attaching chains to an amorphous substrate. The adhesion force between monolayers at a given separation increases monotonically with chain length at full coverage and with coverage for fixed chain length. Friction simulations are performed at shear velocities between 0.02-2 m/s at constant applied pressures between 200 and 600 MPa. Stick-slip motion is observed at full coverage but disappears with disorder. With random defects, the friction becomes insensitive to chain length, defect density, and substrate. PMID- 15525009 TI - Monolayer pattern evolution via substrate strain-mediated spinodal decomposition. AB - Investigations of octylsilane (C8H17SiH3) monolayer pattern formation on Au(111) are reported. Scanning tunneling microscopy data display the evolution of a approximately 6 nm scale pattern of interwoven features concomitant with ejection of surface Au atoms and relaxation of the Au(111) 23xsqrt[3] surface reconstruction. Numerical simulations suggest the surface dynamics are governed by a substrate strain-mediated spinodal decomposition mechanism, novel to organic monolayer formation. Collectively, the experimental and theoretical data indicate strain-inducing Si-Au bond interactions drive the pattern formation and the alkyl chains play a negligible role. PMID- 15525010 TI - Breakdown of classical nucleation theory near isostructural phase transitions. AB - We report simulations of crystal nucleation in binary mixtures of hard spherical colloids with a size ratio of 1:10. The stable crystal phase of this system can be either dense or expanded. We find that, in the vicinity of the solid-solid critical point where the crystallites are highly compressible, small crystal nuclei are less dense than large nuclei. This phenomenon cannot be accounted for by either classical nucleation theory or by the Gibbsian droplet model. We argue that the observed behavior is due to the surface stress of the crystal nuclei. The observed effect highlights a general deficiency of the most frequently used thermodynamic theories for crystal nucleation. Surface stress should lead to an experimentally observable expansion of crystal nuclei of colloids with short ranged attraction and of globular proteins. PMID- 15525011 TI - Proximity effects in self-organized binary particle-block copolymer blends. AB - Dependent on the surface chemistry of gold nanocrystals of equal metal core size, two morphological types of self-organized block copolymer-particle blends are observed: (1) the segregation of the nanocrystals to the interfacial areas or (2) the preferential uniform distribution within one of the respective polymer domains. The confinement of the nanocrystals to the narrow interfacial regions of the microstructure in type one blends results in high local particle filling fractions and gives rise to electromagnetic coupling upon light irradiation, accompanied by a pronounced increase in absorbance. PMID- 15525012 TI - Nonstochastic behavior of atomic surface diffusion on Cu(111) down to low temperatures. AB - Atomic diffusion is usually understood as a succession of random, independent displacements of an adatom over the surface's potential energy landscape. Nevertheless, an analysis of molecular dynamics simulations of self-diffusion on Cu(111) demonstrates the existence of different types of correlations in the atomic jumps at all temperatures. Thus, the atomic displacements cannot be correctly described in terms of a random walk model. This fact has a profound impact on the determination and interpretation of diffusion coefficients and activation barriers. PMID- 15525013 TI - Single-molecule measurements of gold-quenched quantum dots. AB - We report the study of the quenching of quantum dots (CdSe) by gold nanoparticles at the single-molecule level. Double-stranded DNA is used as a rigid spacer to tune the distance between the two nanoparticles. The width of the fluorescent intensity distribution, monitored at different interparticle distances, reflects both the nanoparticle heterogeneity and the fluorescence intermittency of the quantum dot. The fluorescence distribution emitted by single CdSe nanocrystals can easily be distinguished from the fluorescence of partially quenched CdSe. Our results show that the distance-dependence quenching is compatible with a Forster type process. PMID- 15525014 TI - Probing microcavity polariton superfluidity through resonant Rayleigh scattering. AB - We study the motion of a polariton fluid injected into a planar microcavity by a continuous wave laser. In the presence of static defects, the spectrum of the Bogoliubov-like excitations reflects onto the shape and intensity of the resonant Rayleigh scattering emission pattern in both momentum and real space. We find a superfluid regime in which the Rayleigh scattering ring in momentum space collapses as well as its normalized intensity. We show how collective excitation spectra having no analog in equilibrium systems can be observed by tuning the excitation angle and frequency. PMID- 15525015 TI - Phase analysis of quantum oscillations in graphite. AB - The quantum de Haas-van Alphen (dHvA) and Shubnikov-de Haas oscillations measured in graphite were decomposed by pass-band filtering onto contributions from three different groups of carriers. Generalizing the theory of dHvA oscillations for 2D carriers with an arbitrary spectrum and by detecting the oscillation frequencies using a method of two-dimensional phase-frequency analysis which we developed, we identified these carriers as (i) minority holes having a 2D parabolic massive spectrum p(2)(perpendicular)/2m(perpendicular), (ii) massive majority electrons with a 3D spectrum and (iii) majority holes with a 2D Dirac-like spectrum +/ vp(perpendicular) which seems to be responsible for the unusual strongly correlated electronic phenomena in graphite. PMID- 15525016 TI - Real space imaging of one-dimensional standing waves: direct evidence for a Luttinger liquid. AB - Electronic standing waves with two different wavelengths were directly mapped near one end of a single-wall carbon nanotube as a function of the tip position and the sample bias voltage with high-resolution position-resolved scanning tunneling spectroscopy. The observed two standing waves caused by separate spin and charge bosonic excitations are found to constitute direct evidence for a Luttinger liquid. The increased group velocity of the charge excitation, the power-law decay of their amplitudes away from the scattering boundary, and the suppression of the density of states near the Fermi level were also directly observed or calculated from the two different standing waves. PMID- 15525017 TI - Wide energy-window view on the density of states and hole mobility in poly(p phenylene vinylene). AB - Using an electrochemically gated transistor, we achieved controlled and reversible doping of poly(p-phenylene vinylene) in a large concentration range. Our data open a wide energy-window view on the density of states (DOS) and show, for the first time, that the core of the DOS function is Gaussian, while the low energy tail has a more complex structure. The hole mobility increases by more than 4 orders of magnitude when the electrochemical potential is scanned through the DOS. PMID- 15525018 TI - Measurements of Kondo and spin splitting in single-electron transistors. AB - We measure the spin splitting in a magnetic field B of localized states in single electron transistors using a new method, inelastic spin-flip cotunneling. Because it involves only internal excitations, this technique gives the most precise value of the Zeeman energy Delta=/g/mu(B)B. In the same devices we also measure the splitting with B of the Kondo peak in differential conductance. The Kondo splitting appears only above a threshold field as predicted by theory. However, the magnitude of the Kondo splitting at high fields exceeds 2/g/mu(B)B in disagreement with theory. PMID- 15525019 TI - Temperature dependence of spin-transfer-induced switching of nanomagnets. AB - We measure the temperature, magnetic-field, and current dependence for the switching of nanomagnets by a spin-polarized current. Depending on current bias, switching can occur between either two static magnetic states or a static state and a current-driven precessional mode. In both cases, the switching is thermally activated and governed by the sample temperature, not a higher effective magnetic temperature. The activation barriers for switching between static states depend linearly on current, with a weaker dependence for dynamic to static switching. PMID- 15525020 TI - Resonant magnetic excitations at high energy in superconducting YBa2Cu3O6.85. AB - A detailed inelastic neutron scattering study of the high temperature superconductor YBa2Cu3O6.85 provides evidence of new resonant magnetic features, in addition to the well-known resonant mode at 41 meV: (i) a commensurate magnetic resonance peak at 53 meV with an even symmetry under exchange of two adjacent CuO2 layers, and (ii) high-energy incommensurate resonant spin excitations whose spectral weight is around 54 meV. The locus and the spectral weight of these modes provides unrevealed insight about the momentum shape of the electron-hole spin-flip continuum of d-wave superconductors. PMID- 15525021 TI - Precise determination of the orientation of the Dzialoshinskii-Moriya vector in kappa-(BEDT-TTF)2Cu[N(CN)(2)]Cl. AB - A novel electron spin-reorientation transition is discovered by 13C NMR in the quasi-two-dimensional organic antiferromagnet kappa-(BEDT-TTF)(2)Cu[N(CN)(2)]Cl. The spin reorientation occurs as an external field is swept through the orientation of the characteristic vector of the Dzialoshinskii-Moriya (DM) interaction, thus providing a precise determination of the orientation of the DM vector. Such a spin reorientation could help to characterize the DM interaction in other antiferromagnetic systems. PMID- 15525022 TI - Charge ordering and magnetopolarons in Na0.82CoO2. AB - Using spectral ellipsometry, we measured the dielectric function of a Na(0.82(2))CoO2 crystal that exhibits bulk antiferromagnetism with T(N)=19.8 K. We identify two prominent transitions as a function of temperature. The first one at 280 K involves marked changes of the electronic and lattice responses that are indicative of charge ordering in the CoO2 layers. The second transition occurs around T(N)=19.8 K and reveals sizable spin-charge coupling. The data are discussed in terms of charge ordering and formation of magnetopolarons due to a charge-induced spin-state transition of adjacent Co3+ ions. PMID- 15525023 TI - Measurement of the 101Ru-Knight shift of superconducting Sr2RuO4 in a parallel magnetic field. AB - 101Ru-Knight shift (101K) in the spin-triplet superconductor Sr2RuO4 was measured under magnetic fields parallel to the c axis (perpendicular to the RuO2 plane), which is the promising superconducting (SC) d-vector direction in a zero field. We succeeded in measuring K(c) in the field range from 200 to 1200 Oe and at temperatures down to 80 mK, using nuclear-quadrupole-resonance spectra. We found that (101)K(c) is invariant with respect to the field and temperature on passing through H(c2) and T(c) above 200 Oe. This indicates that the spin susceptibility along the c axis does not change in the SC state, at least, in the field greater than 200 Oe. The results imply that the SC d vector is in the RuO2 plane when the magnetic field is applied to the c axis. PMID- 15525024 TI - Importance of the internal shape mode in magnetic vortex dynamics. AB - We investigate the motion of a nonplanar vortex in a circular easy-plane magnet with a rotating in-plane magnetic field. Our numerical simulations of the Landau Lifshitz equations show that the vortex tends to a circular limit trajectory, with an orbit frequency which is lower than the driving field frequency. To describe this we develop a new collective variable theory by introducing additional variables which account for the internal degrees of freedom of the vortex core, strongly coupled to the translational motion. We derive the evolution equations for these collective variables and find limit-cycle solutions whose characteristics are in qualitative agreement with the simulations of the many-spin system. PMID- 15525025 TI - Metallic ferromagnet with square-lattice CoO2 sheets. AB - A single-crystalline film of Sr2CoO4 with square-lattice CoO2 sheets (K2NiF4-type structure) was synthesized, and its electronic properties were characterized. The compound exhibited both ferromagnetic and metallic behaviors, with a fairly high Curie temperature T(C) of approximately 250 K, in contrast to the superconductivity recently found in a triangular-lattice CoO2-sheet compound. The film's large magnetoresistance with current perpendicular to the CoO2 plane showed field-hysteretic behavior analogous to tunneling magnetoresistance. The electronic structure of Sr2CoO4 was also investigated to characterize the quasi two-dimensional ferromagnetic metal state in terms of optical spectroscopy and first-principles calculation. PMID- 15525026 TI - Studying quantum spin systems through entanglement estimators. AB - We study the field dependence of the entanglement of formation in anisotropic S=1/2 antiferromagnetic chains displaying a T=0 field-driven quantum phase transition. The analysis is carried out via quantum Monte Carlo simulations. At zero temperature the entanglement estimators show abrupt changes at and around criticality, vanishing below the critical field, in correspondence with an exactly factorized state, and then immediately recovering a finite value upon passing through the quantum phase transition. At the quantum-critical point, a deep minimum in the pairwise-to-global entanglement ratio shows that multispin entanglement is strongly enhanced; moreover this signature represents a novel way of detecting the quantum phase transition of the system, relying entirely on entanglement estimators. PMID- 15525027 TI - Dipolar spin correlations in classical pyrochlore magnets. AB - We study spin correlations for the highly frustrated classical pyrochlore lattice antiferromagnets with O(N) symmetry in the limit T-->0. We conjecture that a local constraint obeyed by the extensively degenerate ground states dictates a dipolar form for the asymptotic spin correlations, at all N not equal 2 for which the system is paramagnetic down to T=0. We verify this conjecture in the cases N=1 and N=3 by simulations and to all orders in the 1/N expansion about the solvable N=infinity limit. Remarkably, the N=infinity formulas are an excellent fit, at all distances, to the correlators at N=3 and even at N=1. Thus we obtain a simple analytical expression also for the correlations of the equivalent models of spin ice and cubic water ice, Ic. PMID- 15525028 TI - Field-induced orbital and magnetic phases in Ca3Ru2O7. AB - Magnetic-field- and temperature-dependent Raman scattering studies of Ca3Ru2O7 reveal dramatic field-dependent properties arising from transitions between various complex orbital and magnetic phases, including a field-induced orbital ordered to orbital-disordered transition (H(o) // hard axis), and a reentrant orbital-ordered to orbital-disordered to orbital-ordered transition (H(o) // easy axis). We find that the dramatic magnetic-field properties are most prevalent in a "mixed"-magnetic and -orbital phase regime, providing evidence for a strong connection between orbital phase inhomogeneity and "colossal" field effects in the ruthenates. PMID- 15525029 TI - Origin of the glassy magnetic behavior of the phase segregated state of the perovskites. AB - In this Letter we demonstrate that the phase segregated state observed in many rare-earth perovskites constitutes a sort of self-generated assembly of magnetic clusters in which magnetic interaction introduces collectivity among them. We show that the observed glassy behavior (memory, aging, etc.) can be perfectly understood taking into account only the intercluster interactions. We address the fundamental question about whether this state constitutes classical spin glass or if, on the other hand, a new universality class must be defined. PMID- 15525030 TI - Measuring the uniaxial strain of individual single-wall carbon nanotubes: resonance Raman spectra of atomic-force-microscope modified single-wall nanotubes. AB - Raman spectroscopy is used to measure the strain in individual single-wall carbon nanotubes, strained by manipulation with an atomic-force-microscope tip. Under strains varying from 0.06%-1.65%, the in-plane vibrational mode frequencies are lowered by as much as 1.5% (40 cm(-1)), while the radial breathing mode (RBM) remains unchanged. The RBM Stokes/anti-Stokes intensity ratio remains unchanged under strain. The elasticity of these strain deformations is demonstrated as the down-shifted Raman modes resume their prestrain frequencies after a nanotube is broken under excessive strain. PMID- 15525031 TI - Monitoring surface charge movement in single elongated semiconductor nanocrystals. AB - We demonstrate a universal correlation between the spectral linewidth and position of the excitonic transition in the spectral jitter observed from single elongated colloidal quantum dots. Breaking the symmetry of electron and hole confinement as well as of the spatial directions for surface charge diffusion enables us to microscopically track meandering surface charges, providing a novel probe of the particle's nanoenvironment. Spectral diffusion exhibits only a weak temperature dependence, which allows us to uncover the single particle homogeneous linewidth of 50 meV at room temperature. PMID- 15525032 TI - 23Na NMR evidence for charge order and anomalous magnetism in NaxCoO2. AB - Oriented powder samples of NaxCoO2 are studied by 23Na NMR and SQUID magnetometry. In nominal 0.50. It is found that this covalent/quantum contribution is determined nonlocally by the topological structure in the Q--> space. The condition for the charge pumping for a cyclic change of Q--> is also obtained. Applications of this picture to various organic ferroelectrics and BaTiO3 are discussed. PMID- 15525034 TI - Long-range interlayer interactions in ferroelectric liquid crystals. AB - Some smectic liquid crystals exhibit a series of phases, including ferroelectric, antiferroelectric, and ferrielectric commensurate structures as well as an incommensurate phase. A long-standing problem has been to understand the origin of the long-range interaction responsible for this rich variety of phases. We study a model that incorporates thermal fluctuations in the flexing of layers and find that it supports commensurate and incommensurate structures. The vibrational entropy competes with an assumed helical interaction between nearest-neighbor layers. An increase in temperature then leads to an unwinding of the helix that proceeds at first through commensurate phases and then into an incommensurate phase. This result is consistent with the experimentally observed "distorted clock model." PMID- 15525035 TI - Inaccessibility in online learning of recurrent neural networks. AB - We apply nonlinear dynamical system techniques to recurrent neural networks. In particular, we numerically analyze the dynamical system characteristics of the online learning process. By introducing the notion of inaccessibility, we show that the learning process is well characterized by strong nonhyperbolicity and inaccessibility, which is a greater uncertainty than chaotic unpredictability. These results are clearly contrasted with a gradient descent dynamics, or ordinary chaos. PMID- 15525036 TI - Viscoelasticity of single wall carbon nanotube suspensions. AB - We investigate the viscoelastic properties of an associating rigid rod network: aqueous suspensions of surfactant stabilized single wall carbon nanotubes (SWNTs). The SWNT suspensions exhibit a rigidity percolation transition with an onset of solidlike elasticity at a volume fraction of 0.0026; the percolation exponent is 2.3+/-0.1. At large strain, the solidlike samples show volume fraction dependent yielding. We develop a simple model to understand these rheological responses and show that the shear dependent stresses can be scaled onto a single master curve to obtain an internanotube interaction energy per bond approximately 40k(B)T. Our experimental observations suggest SWNTs in suspension form interconnected networks with bonds that freely rotate and resist stretching. Suspension elasticity originates from bonds between SWNTs rather than from the stiffness or stretching of individual SWNTs. PMID- 15525037 TI - Non-Tikhonov asymptotic properties of cardiac excitability. AB - Models of electric excitability of cardiac cells can be studied by singular perturbation techniques. To do this one should take into account parameters appearing in equations in nonstandard ways. The physical reason for this is near perfect switch behavior of ionic current gates. This leads to a definition of excitability different from the currently accepted one. The asymptotic structure revealed by our analysis can be used to devise simplified caricature models, to obtain approximate analytical solutions, and to facilitate numerical simulations. PMID- 15525038 TI - Minimalist molecular model for nanopore selectivity. AB - Using a simple model it is shown that the cost of constraining a hydrated potassium ion inside a narrow nanopore is smaller than the cost of constraining the smaller hydrated sodium ion. The former allows for a greater distortion of its hydration shell and can therefore maintain a better coordination. We propose that in this way the larger ion can go through narrow pores more easily. This is relevant to the molecular basis of ion selective nanopores and since this mechanism does not depend on the molecular details of the pore, it could also operate in all sorts of nanotubes, from biological to synthetic. PMID- 15525040 TI - Locked-floating-solid to locked-smectic transition in colloidal systems. AB - We investigate two-dimensional melting of a colloidal system in the presence of a one-dimensional periodic substrate potential created by two interfering laser beams. We study the commensurability ratio p=sqrt[3]a/2d=2 with a the mean particle distance and d the period of the periodic potential. In contrast with the previously investigated case p=1, here we observe that melting of the locked floating solid occurs via a novel locked-smectic phase, predicted by recent theoretical studies. PMID- 15525039 TI - Human hand moves proactively to the external stimulus: an evolutional strategy for minimizing transient error. AB - We investigated particularly the proactive nature of the visual-motor system by steady and transient experiments of a hand-tracking task, and confirmed that the hand motion precedes on the average the target motion in steady runs within a finite frequency range of the sinusoidal target motion. The question why and how much the hand motion should precede was answered by frequency-jump experiments. The results implied that the positive phase shift of the hand motion represents the proactive nature of the visual-motor control system which is adaptationally developed for each person to minimize the transient error of the hand motion when the target motion changes unexpectedly. PMID- 15525041 TI - Continuous monitoring of Mg oxidation by internal exoemission. AB - Thin-film Mg/Si(111) Schottky diodes are exposed to oxygen to detect chemicurrents in the devices. The detected charge is created by nonadiabatic energy dissipation and due to either internal exoemission currents or surface chemiluminescence induced photocurrents. Both contributions can be distinguished by changing the metal film thickness of the device. Auger electron spectroscopy to study the oxygen uptake demonstrates that the chemicurrent transients represent truly the time dependent reaction rate at the surface. Model calculations indicate that the current monitors Mg oxide island nucleation and growth. PMID- 15525042 TI - Pinning force in active media. AB - Pinning of vortices by defects plays an important role in various physical (superconductivity, superfluidity, etc.) or biological (propagation in cardiac muscle) situations. Which defects act as pinning centers? We propose a way to study this general problem by using an advection field to quantify the attraction between an obstacle and a vortex. A full solution is obtained for the real Ginzburg-Landau equation (RGLE). Two pinning mechanisms are found in excitable media. Our results suggest strong analogies with the RGLE when the heterogeneity is excitable. Unpinning from an unexcitable obstacle is qualitatively harder, resulting in a stronger pinning force. We discuss the implications of our results to control vortices and propose experiments in a chemical active medium and in cardiac tissue. PMID- 15525043 TI - Geographical coarse graining of complex networks. AB - We perform a renormalization-grouplike numerical analysis of geographically embedded complex networks on a two-dimensional square lattice. At each step of the coarse-graining procedure, the four vertices on each 2x2 square box are merged to a single vertex, resulting in a coarse-grained system of smaller size. Repetition of the process leads to the observation that the coarse-graining procedure does not alter the qualitative characteristics of the original scale free network, which opens the possibility of subtracting a smaller network from the original network without destroying the important structural properties. The implication of the result is also suggested in the context of the recent study of the human brain functional network. PMID- 15525044 TI - Low-energy excitations in resonant inelastic X-Ray scattering of alpha(')-NaV2O5. PMID- 15525046 TI - Comment on "Direct measurement of the spin polarization of the magnetic semiconductor (Ga,Mn)As". PMID- 15525048 TI - Fraying of fractional statistics at the edge. PMID- 15525050 TI - Local molecular dynamics with coulombic interactions. AB - We propose a local, O(N) molecular dynamics algorithm for the simulation of charged systems. The long ranged Coulomb potential is generated by a propagating electric field that obeys modified Maxwell equations. On coupling the electrodynamic equations to an external thermostat we show that the algorithm produces an effective Coulomb potential between particles. On annealing the electrodynamic degrees of freedom the field configuration converges to a solution of the Poisson equation much like the electronic degrees of freedom approach the ground state in ab initio molecular dynamics. PMID- 15525051 TI - Quantum monte carlo algorithm based on two-body density functional theory for fermionic many-body systems: application to 3He. AB - We construct a quantum Monte Carlo algorithm for interacting fermions using the two-body density as the fundamental quantity. The central idea is mapping the interacting fermionic system onto an auxiliary system of interacting bosons. The correction term is approximated using correlated wave functions for the interacting system, resulting in an effective potential that represents the nodal surface. We calculate the properties of 3He and find good agreement with experiment and with other theoretical work. In particular, our results for the total energy agree well with other calculations where the same approximations were implemented but the standard quantum Monte Carlo algorithm was used. PMID- 15525052 TI - Unified derivation of tunneling times from decoherence functionals. AB - The four tunneling times, the Larmor time, the Buttiker-Landauer time, the Bohm Wigner time, and the Pollak-Miller time, originally obtained from very different physical models, are derived in a unified manner from the Gell-Mann-Hartle decoherence functionals. The origin of the two types of derivatives in the expressions for these tunneling times is clarified at the level of Feynman paths. PMID- 15525053 TI - Spontaneous macroscopic spin polarization in independent spinor Bose-Einstein condensates. AB - We present experimental evidence for the spontaneous formation of a macroscopic spin polarization in overlapping regions of two independent Bose-Einstein condensates produced in different hyperfine states of 87Rb. The condensates are independent in the sense that we do not explicitly introduce a relative phase between them. A single "spin-tip" pulse maps the transverse spin polarization into longitudinal spin polarization, and the atomic density distributions are measured with a Stern-Gerlach imaging method. The resulting matter-wave interference patterns are anticorrelated. PMID- 15525054 TI - Atom-dimer scattering for confined ultracold fermion gases. AB - We solve the three-body problem of a quasi-one-dimensional ultracold Fermi gas with parabolic confinement length a (perpendicular) and 3D scattering length a. On the two-body level, there is a Feshbach-type resonance at a (perpendicular)/a approximately 1.46, and a dimer state for arbitrary a (perpendicular)/a. The three-body problem is shown to be universal, and described by the atom-dimer scattering length a(ad) and a range parameter b(ad). In the dimer limit a (perpendicular)/a>>1, we find a repulsive zero-range atom-dimer interaction. For a (perpendicular)/a<<-1, however, the potential has long range, with a(ad)>0 and b(ad)>>a(ad). There is no trimer state, and despite a(ad)=0 at a( perpendicular)/a approximately 2.6, there is no resonance enhancement of the interaction. PMID- 15525055 TI - Experimental purification of single qubits. AB - We report the experimental realization of the purification protocol for single qubits sent through a depolarizing channel. The qubits are associated with polarization states of single photons and the protocol is achieved by means of passive linear optical elements. The present approach may represent a convenient alternative to the distillation and error correction protocols of quantum information. PMID- 15525057 TI - Distillation protocols: output entanglement and local mutual information. AB - A complementary behavior between local mutual information and average output entanglement is derived for arbitrary bipartite ensembles. This leads to bounds on the yield of entanglement in distillation protocols that involve disinguishing. This bound is saturated in the hashing protocol for distillation, for Bell-diagonal states. PMID- 15525056 TI - Security of quantum key distribution with coherent states and homodyne detection. AB - We assess the security of a quantum key distribution protocol relying on the transmission of Gaussian-modulated coherent states and homodyne detection. This protocol is shown to be equivalent to an entanglement purification protocol using CSS codes followed by key extraction, and is thus secure against any eavesdropping strategy. PMID- 15525058 TI - Quantum cryptography without switching. AB - We propose a new coherent state quantum key distribution protocol that eliminates the need to randomly switch between measurement bases. This protocol provides significantly higher secret key rates with increased bandwidths than previous schemes that only make single quadrature measurements. It also offers the further advantage of simplicity compared to all previous protocols which, to date, have relied on switching. PMID- 15525059 TI - Efficient simulations of gas-grain chemistry in interstellar clouds. AB - Chemical reactions on dust grains are of crucial importance in interstellar chemistry because they produce molecular hydrogen and various organic molecules. Because of the submicron size of the grains and the low flux, the surface populations of reactive species are small and strongly fluctuate. Under these conditions rate equations fail and the master equation is needed for modeling these reactions. However, the number of equations grows exponentially with the number of reactive species, severely limiting its feasibility. Here we present a method which dramatically reduces the number of equations, thus enabling the incorporation of the master equation in models of interstellar chemistry. PMID- 15525060 TI - Eigenmodes and thermodynamics of a Coulomb chain in a harmonic potential. AB - The density of ions trapped in a harmonic potential in one dimension is not uniform. Consequently the eigenmodes are not phononlike waves. We calculate the long-wavelength modes in the continuum limit, and evaluate the density of states in the short-wavelength limit for chains of N>>1 ions. Remarkably, the results that are found analytically in the thermodynamic limit provide a good estimate of the spectrum of excitations of small chains down to few tens of ions. The spectra are used to compute the thermodynamic functions of the chain. Deviations from the extensivity of the thermodynamic quantities are found. An analytic expression for the critical transverse frequency determining the stability of a linear chain is derived. PMID- 15525061 TI - Limit on the present temporal variation of the fine structure constant. AB - The comparison of different atomic transition frequencies over time can be used to determine the present value of the temporal derivative of the fine structure constant alpha in a model-independent way without assumptions on constancy or variability of other parameters, allowing tests of the consequences of unification theories. We have measured an optical transition frequency at 688 THz in 171Yb+ with a cesium atomic clock at 2 times separated by 2.8 yr and find a value for the fractional variation of the frequency ratio f(Yb)/f(Cs) of (-1.2+/ 4.4)x10(-15) yr(-1), consistent with zero. Combined with recently published values for the constancy of other transition frequencies this measurement sets an upper limit on the present variability of alpha at the level of 2.0x10(-15) yr( 1) (1sigma), corresponding so far to the most stringent limit from laboratory experiments. PMID- 15525063 TI - Antineutrino spectroscopy with large water Cerenkov detectors. AB - We propose modifying large water C erenkov detectors by the addition of 0.2% gadolinium trichloride, which is highly soluble, newly inexpensive, and transparent in solution. Since Gd has an enormous cross section for radiative neutron capture, with summation operatorE(gamma)=8 MeV, this would make neutrons visible for the first time in such detectors, allowing antineutrino tagging by the coincidence detection reaction nu (e)+p-->e(+)+n (similarly for nu (mu)). Taking Super-Kamiokande as a working example, dramatic consequences for reactor neutrino measurements, first observation of the diffuse supernova neutrino background, galactic supernova detection, and other topics are discussed. PMID- 15525062 TI - Direct observation of charge inversion by multivalent ions as a universal electrostatic phenomenon. AB - We have directly observed reversal of the polarity of charged surfaces in water upon the addition of trivalent and quadrivalent ions using atomic force microscopy. The bulk concentration of multivalent ions at which charge inversion reversibly occurs depends only very weakly on the chemical composition, surface structure, size, and lipophilicity of the ions, but is very sensitive to their valence. These results support the theoretical proposal that spatial correlations between ions are the driving mechanism behind charge inversion. PMID- 15525064 TI - Rotating black holes in higher dimensions with a cosmological constant. AB - We present the metric for a rotating black hole with a cosmological constant and with arbitrary angular momenta in all higher dimensions. The metric is given in both Kerr-Schild and the Boyer-Lindquist form. In the Euclidean-signature case, we also obtain smooth compact Einstein spaces on associated S(D-2) bundles over S2, infinitely many for each odd D>/=5. Applications to string theory and M theory are indicated. PMID- 15525065 TI - 60Fe anomaly in a deep-sea manganese crust and implications for a nearby supernova source. AB - A nearby supernova (SN) explosion in the past can be confirmed by the detection of radioisotopes on Earth that were produced and ejected by the SN. We have now measured a well resolved time profile of the 60Fe concentration in a deep-sea ferromanganese crust and found a highly significant increase 2.8 Myr ago. The amount of 60Fe is compatible with the deposition of ejecta from a SN at a distance of a few 10 pc. The well defined time of the SN explosion makes it possible to search for plausible correlations with other events in Earth's history. PMID- 15525066 TI - Chameleon fields: awaiting surprises for tests of gravity in space. AB - We present a novel scenario where a scalar field acquires a mass which depends on the local matter density: the field is massive on Earth, where the density is high, but is essentially free in the solar system, where the density is low. All existing tests of gravity are satisfied. We predict that near-future satellite experiments could measure an effective Newton's constant in space different from that on Earth, as well as violations of the equivalence principle stronger than currently allowed by laboratory experiments. PMID- 15525067 TI - J/Psi production in pp collisions at square root = 200 GeV at the BNL relativistic heavy ion collider. AB - We study J/psi production in pp collisions at BNL Relativistic Heavy Ion Collider (RHIC) within the PHENIX detector acceptance range using the color singlet and color octet mechanism which are based on perturbative QCD and nonrelativistic QCD. Here we show that the color octet mechanism reproduces the RHIC data for J/psi production in pp collisions with respect to the p(T) distribution, the rapidity distribution, and the total cross section at square root = 200 GeV. The color singlet mechanism leads to a relatively small contribution to the total cross section when compared to the octet contribution. PMID- 15525068 TI - New half-life measurement of 182Hf: improved chronometer for the early solar system. AB - The decay of 182Hf, now extinct, into stable 182W has developed into an important chronometer for studying early solar system processes such as the accretion and differentiation of planetesimals and the formation of the Earth and the Moon. The only 182Hf half-life measurements available were performed 40 years ago and resulted in an imprecise half-life of (9+/-2)x10(6) yr. We redetermined the half life by measuring the specific activity of 182Hf based on two independent methods, resulting in a value of t(1/2)(182Hf)=(8.90+/-0.09)x10(6) yr, in good agreement with the previous value, but with a 20 times smaller uncertainty. The greatly improved precision of this half-life now permits very precise intercalibration of the 182Hf-182W isotopic system with other chronometers. PMID- 15525069 TI - Chiral bands, dynamical spontaneous symmetry breaking, and the selection rule for electromagnetic transitions in the chiral geometry. AB - A model for a special configuration in triaxial odd-odd nuclei is constructed which exhibits degenerate chiral bands with a sizable rotation, a manifestation of dynamical spontaneous symmetry breaking. A quantum number obtained from the invariance of the model Hamiltonian, which characterizes observable states, is given and selection rules for electromagnetic transition probabilities in chiral bands is derived in terms of this quantum number. The degeneracy of the lowest two bands is indeed obtained in the numerical diagonalization of the Hamiltonian at an intermediate spin range, over which electromagnetic transitions follow exactly the selection rule expected for the chiral geometry. PMID- 15525070 TI - Lack of Hohenberg-Kohn theorem for excited states. AB - For a given excited state there exist densities that arise from more than one external potential. This is due to a qualitatively different energy-density relationship from that of the ground state and is related to positive eigenvalues in the nonlocal susceptibility for excited states. Resulting problems with the generalization of the density functional methodology to excited states are discussed. PMID- 15525072 TI - Mixed quantum-classical treatment of vibrational decoherence. AB - The mixed quantum-classical method based on the Bohmian formulation of quantum mechanics [J. Chem. Phys. 113, 9369 (2000)]] is applied to study the process of vibrational decoherence of I2 in a dense helium environment. Specifically, the revival of vibrational wave packets, detectable by pump-probe spectroscopy, is a quantum phenomena which depends sensitively on the coherence between the vibrational levels excited by the pump pulse. The time-dependent pump-probe revival signal is a very sensitive way of detecting vibrational dephasing induced by an environment. The very good agreement between previous experimental signals and calculations presented in ths work confirms the theoretical approach and provides a promising basis for the prediction and interpretation of future experiments exploiting quantum revivals as a probe of decoherence. PMID- 15525071 TI - Can a quantum nondemolition measurement improve the sensitivity of an atomic magnetometer? AB - We consider the limitations due to noise (e.g., quantum projection noise and photon shot-noise) on the sensitivity of an idealized atomic magnetometer that utilizes spin squeezing induced by a continuous quantum nondemolition measurement. Such a magnetometer measures spin precession of N atomic spins by detecting optical rotation of far-detuned light. We show that for very short measurement times, the optimal sensitivity scales as N(-3/4); if strongly squeezed probe light is used, the Heisenberg limit of N-1 scaling can be achieved. However, if the measurement time exceeds tau(rel)/N(1/2) in the former case, or tau(rel)/N in the latter, where tau(rel) is the spin relaxation time, the scaling becomes N(-1/2), as for a standard shot-noise-limited magnetometer. PMID- 15525073 TI - Imaging of s and d partial-wave interference in quantum scattering of identical bosonic atoms. AB - We report on the direct imaging of s and d partial-wave interference in cold collisions of atoms. Two ultracold clouds of 87Rb atoms were accelerated by magnetic fields to collide at energies near a d-wave shape resonance. The resulting halos of scattered particles were imaged using laser absorption. By scanning across the resonance we observed a marked evolution of the scattering patterns due to the energy dependent phase shifts for the interfering s and d waves. Since only two partial-wave states are involved in the collision process the scattering yield and angular distributions have a simple interpretation in terms of a theoretical model. PMID- 15525074 TI - Interferometric determination of the s and d-wave scattering amplitudes in 87Rb. AB - We demonstrate an interference method to determine the low-energy elastic scattering amplitudes of a quantum gas. We linearly accelerate two ultracold atomic clouds up to energies of 1.2 mK and observe the collision halo by direct imaging in free space. From the interference between s- and d- partial waves in the differential scattering pattern we extract the corresponding phase shifts. The method does not require knowledge of the atomic density. This allows us to infer accurate values for the s- and d-wave scattering amplitudes from the zero energy limit up to the first Ramsauer minimum using only the van der Waals C6 coefficient as theoretical input. For the 87Rb triplet potential, the method reproduces the scattering length with an accuracy of 6%. PMID- 15525076 TI - Evaluation of two-photon nonlinearity by a semiclassical method. AB - In order to discuss the two-photon nonlinearity theoretically, both photons and nonlinear materials should be treated quantum mechanically, which usually is a heavy theoretical task. Contrarily, nonlinear optics for classical light has been developed well and a detailed analysis is possible for realistic complex nonlinear systems. Here we show that the two-photon nonlinearity can be evaluated from the linear and third-order nonlinear output fields against a classical input pulse, which contains 2(-1/2) photons on average. PMID- 15525075 TI - Femtosecond interatomic Coulombic decay in free neon clusters: large lifetime differences between surface and bulk. AB - A quantitative determination of 2s vacancy lifetimes in surface and bulk atoms of free Ne clusters has been made. While for free atoms the 2s inner-valence hole has a ps lifetime, it reduces to 6+/-1 fs for cluster bulk atoms. For surface atoms, the lifetime is on average longer than 30 fs. The lifetime estimate was obtained from fits of high-resolution photoelectron spectra of Ne clusters. The shortening of the lifetime is attributed to the coordination dependent interatomic Coulombic decay, which is extremely sensitive to internuclear distances. PMID- 15525077 TI - Connection between phase singularities and the radiation pattern of a slit in a metal plate. AB - We report a new fundamental relation between the minima of the far-zone radiation pattern of a narrow slit in a metal plate and the location of phase singularities in the intermediate field. If a system parameter such as the wavelength is changed, a previously unappreciated singular optics phenomenon occurs: namely, the transition of a near-zone phase singularity into a singularity of the radiation pattern. Our results have significance for the design of novel nanoscale light sources and antennas. PMID- 15525078 TI - Self-compression of ultrashort pulses through ionization-induced spatiotemporal reshaping. AB - We present the first demonstration of a new mechanism for temporal compression of ultrashort light pulses that operates at high (i.e., ionizing) intensities. By propagating pulses inside a hollow waveguide filled with low-pressure argon gas, we demonstrate a self-compression from 30 to 13 fs, without the need for any external dispersion compensation. Theoretical models show that 3D spatiotemporal reshaping of the pulse due to a combination of ionization-induced spectral broadening, plasma-induced refraction, and guiding in the hollow waveguide are necessary to explain the compression mechanism. PMID- 15525079 TI - Time reversal of light with linear optics and modulators. AB - We introduce a new physical process that can perform a complete time-reversal operation on any electromagnetic pulse. The process uses only small refractive index modulations of linear optical elements. No nonlinear multiphoton effects such as four-wave mixing are required. The introduced process can be implemented on chip with standard semiconductor materials. Furthermore, the same process can be used to compress or expand the spectrum of electromagnetic waves while completely preserving the coherent information. We exhibit the time-reversal process by first-principles simulations of microcavity complexes in photonic crystals. PMID- 15525080 TI - Experimental relevance of global properties of time-delayed feedback control. AB - We show by means of theoretical considerations and electronic circuit experiments that time-delayed feedback control suffers from severe global constraints if transitions at the control boundaries are discontinuous. Subcritical behavior gives rise to small basins of attraction and thus limits the control performance. The reported properties are, on the one hand, universal since the mechanism is based on general arguments borrowed from bifurcation theory and, on the other hand, directly visible in experimental time series. PMID- 15525081 TI - Chimera states for coupled oscillators. AB - Arrays of identical oscillators can display a remarkable spatiotemporal pattern in which phase-locked oscillators coexist with drifting ones. Discovered two years ago, such "chimera states" are believed to be impossible for locally or globally coupled systems; they are peculiar to the intermediate case of nonlocal coupling. Here we present an exact solution for this state, for a ring of phase oscillators coupled by a cosine kernel. We show that the stable chimera state bifurcates from a spatially modulated drift state, and dies in a saddle-node bifurcation with an unstable chimera state. PMID- 15525082 TI - Temporal chaos versus spatial mixing in reaction-advection-diffusion systems. AB - We develop a theory describing the transition to a spatially homogeneous regime in a mixing flow with a chaotic in time reaction. The transverse Lyapunov exponent governing the stability of the homogeneous state can be represented as a combination of Lyapunov exponents for spatial mixing and temporal chaos. This representation, being exact for time-independent flows and equal Pe clet numbers of different components, is demonstrated to work accurately for time-dependent flows and different Pe clet numbers. PMID- 15525083 TI - Three-species collisionless reconnection: effect of O+ on magnetotail reconnection. AB - The nature of collisionless reconnection in a three-species plasma composed of a heavy species, protons, and electrons is examined. In addition to the usual two length scales present in two-species reconnection, there are two additional larger length scales in the system: one associated with a "heavy whistler" which produces a large scale quadrupolar out-of-plane magnetic field, and one associated with the "heavy Alfven" wave which can slow the outflow speed and thus the reconnection rate. The consequences for reconnection with O+ present in the magnetotail are discussed. PMID- 15525084 TI - Hamiltonian description of low-temperature relativistic plasmas. AB - We develop a low-temperature fluidlike plasma model without recourse to a collisional closure. The equations are closed by treating the momentum spread asymptotically. This model inherits the Hamiltonian structure, including Casimir invariants of the Vlasov-Maxwell theory. We study temperature evolution in the wake of an intense laser pulse propagating in a plasma. We show that the momentum spread is intrinsically anisotropic and that, for conditions corresponding to recent experiments, modest heating occurs. PMID- 15525085 TI - Hall magnetic reconnection rate. AB - Two-dimensional Hall magnetohydrodynamic simulations are used to determine the magnetic reconnection rate in the Hall limit. The simulations are run until a steady state is achieved for four initial current sheet thicknesses: L=1,5,10, and 20c/omega(pi), where c/omega(pi) is the ion inertial length. It is found that the asymptotic (i.e., time independent) state of the system is nearly independent of the initial current sheet width. Specifically, the Hall reconnection rate is weakly dependent on the initial current layer width and is partial differential Phi/ partial differential t less, similar 0.1V(A0)B0, where Phi the reconnected flux, and V(A0) and B0 are the Alfven velocity and magnetic field strength in the upstream region. Moreover, this rate appears to be independent of the scale length on which the electron "frozen-in" condition is broken (as long as it is /=0.2 single crystals, Deltarho(anis)(ab)(theta) displays a deviation from the typical quasiparticle contribution (proportional, sin((2)theta) for temperatures smaller than a certain value T(phi) in the pseudogap region. This deviation is consistent with a flux-flow type contribution to angular magnetoresistivity, indicating the presence of vortexlike excitations above the zero-field critical temperature in the pseudogap region. PMID- 15525116 TI - Nondestructive readout for a superconducting flux qubit. AB - We present a new readout method for a superconducting flux qubit, based on the measurement of the Josephson inductance of a superconducting quantum interference device that is inductively coupled to the qubit. The intrinsic flux detection efficiency and backaction are suitable for a fast and nondestructive determination of the quantum state of the qubit, as needed for readout of multiple qubits in a quantum computer. We performed spectroscopy of a flux qubit and we measured relaxation times of the order of 80 micros. PMID- 15525117 TI - Fermi surface topology of Ca1.5Sr0.5RuO4 determined by angle-resolved photoelectron spectroscopy. AB - We report angle-resolved photoelectron spectroscopy results of the Fermi surface of Ca1.5Sr0.5RuO4, which is at the boundary of magnetic/orbital instability in the phase diagram of the Ca-substituted Sr ruthenates. Three t(2g) energy bands and the corresponding Fermi surface sheets are observed, which are also present in the Ca-free Sr2RuO4. We find that while the Fermi surface topology of the alpha,beta (d(yz,zx)) sheets remains almost the same in these two materials, the gamma (d(xy)) sheet exhibits a holelike Fermi surface in Ca1.5Sr0.5RuO4 in contrast to being electronlike in Sr2RuO4. Our observation of all three volume conserving Fermi surface sheets clearly demonstrates the absence of orbital selective Mott transition, which was proposed theoretically to explain the unusual transport and magnetic properties in Ca1.5Sr0.5RuO4. PMID- 15525118 TI - Unusual directional dependence of exchange energies in GaAs diluted with Mn: is the RKKY description relevant? AB - Ferromagnetism in Mn-doped GaAs, the prototypical dilute magnetic semiconductor (DMS), has so far been attributed to hole mediated RKKY-type interactions. First principles calculations reveal a strong direction dependence of the ferromagnetic (FM) stabilization energy for Mn pairs, a dependence that cannot be explained within RKKY. In the limit of a hostlike hole engineered here where the RKKY model is applicable, the exchange energies are strongly reduced, suggesting that this limit cannot explain the observed ferromagnetism. The dominant contribution stabilizing the FM state is found to be maximal for 110-oriented Mn pairs and minimal for 100-oriented Mn pairs, providing an alternate explanation for magnetism in such materials in terms of energy lowering due to p-d hopping interactions, and offering a new design degree of freedom to enhance FM. PMID- 15525119 TI - Thermal conductivity of geometrically frustrated, ferroelectric YMnO3: extraordinary spin-phonon interactions. AB - The thermal conductivity of the magnetically frustrated, ferroelectric YMnO3 exhibits an isotropic suppression in the cooperative paramagnetic state, followed by a sudden increase upon magnetic ordering. This unprecedented behavior without an associated static structural distortion probably originates from the strong dynamic coupling between acoustic phonons and low-energy spin fluctuations in geometrically frustrated magnets. The replacement of magnetic Ho for Y at the ferroelectrically active site results in an even larger effect, suggestive of the strong influence of multiferroicity. PMID- 15525120 TI - Diffusive transport in spin-1 chains at high temperatures. AB - We present a numerical study on the spin and thermal conductivities of the spin-1 Heisenberg chain in the high temperature limit, in particular, the Drude weight contribution and frequency dependence. We use the exact diagonalization and the recently developed microcanonical Lanczos method; it allows us a finite size scaling analysis by the study of significantly larger lattices. This work, pointing to a diffusive rather than ballistic behavior, is discussed with respect to other recent theoretical and experimental studies. PMID- 15525121 TI - Spin order in one-dimensional Kondo and Hund lattices. AB - We study numerically the one-dimensional Kondo and Hund lattices consisting of localized spins interacting antiferromagnetically or ferromagnetically with the itinerant electrons, respectively. Using the density-matrix renormalization group we find, for both models and in the small coupling regime, the existence of new magnetic phases where the local spins order forming ferromagnetic islands coupled antiferromagnetically. Furthermore, by increasing the interaction parameter |J| we find that this order evolves toward the ferromagnetic regime through a spiral like phase with longer characteristic wavelengths. These results shed new light on the zero temperature magnetic phase diagram for these models. PMID- 15525122 TI - Monolayer resolved oscillating hyperfine fields in epitaxial face-centered tetragonal Co(001) films. AB - Ultrahigh-quality thin fct-Co films grown on Cu(001) have been investigated by 59Co nuclear magnetic resonance. The influence of the spin-dependent electron scattering at the interfaces is observed for at least four Co atomic layers from the interface with monolayer resolution. An oscillatory effect on the Co hyperfine field with a period of several monolayers is measured, corresponding to the oscillating conduction electron polarization. The observation is exclusively possible in this system due to its very narrow resonance lines, corresponding to a virtually perfect Co structure. PMID- 15525123 TI - Anisotropic ferromagnetism in substituted zinc oxide. AB - Room-temperature ferromagnetism is observed in (110) oriented ZnO films made from targets containing 5 at. % of Sc, Ti, V, Fe, Co, or Ni, but not Cr, Mn, or Cu ions. There are large moments, 2.6 micro(B) and 0.5 micro(B)/dopant atom for Co- and Ti-containing oxides, respectively. There is also a moment of 0.3 micro(B)/Sc. Magnetization is very anisotropic, with variations of up to a factor of 3 depending on the orientation of the applied field relative to the substrate. Results are interpreted in terms of a spin-split donor impurity-band model, which can account for ferromagnetism in insulating or conducting high-k oxides with concentrations of magnetic ions that lie far below the percolation threshold. Magnetic moments are associated with two-electron defects in the films as well as unpaired electrons of the 3d ions. PMID- 15525124 TI - Chirality distribution and transition energies of carbon nanotubes. AB - From resonant Raman scattering on isolated nanotubes we obtained the optical transition energies, the radial breathing mode frequency, and the Raman intensity of both metallic and semiconducting tubes. We unambiguously assigned the chiral index (n(1),n(2)) of approximately 50 nanotubes based solely on a third-neighbor tight-binding Kataura plot and find omega(RBM)=(214.4+/-2) cm(-1) nm/d+(18.7+/-2) cm(-1). In contrast to luminescence experiments we observe all chiralities including zigzag tubes. The Raman intensities have a systematic chiral-angle dependence confirming recent ab initio calculations. PMID- 15525125 TI - Structural anomalies and multiferroic behavior in magnetically frustrated TbMn2O5. AB - We have studied the magnetostructural phase diagram of multiferroic TbMn2O5 as a function of temperature and magnetic field by neutron diffraction. Dielectric and magnetic anomalies are found to be associated with steps in the magnetic propagation vector, including a rare example of a commensurate-incommensurate transition on cooling below 24 K, and in the structural parameters. The geometrically frustrated magnetic structure is stabilized by "canted antiferroelectric" displacements of the Mn3+ ions, an example of the magnetic Jahn-Teller effect. The Tb moments order ferromagnetically at low temperatures in an applied field, while the Mn magnetic structure is largely unchanged. PMID- 15525126 TI - Size-dependent optical properties of VO2 nanoparticle arrays. AB - The size effects on the optical properties of vanadium dioxide nanoparticles in ordered arrays have been studied. Contrary to previous VO2 studies, we observe that the optical contrast between the semiconducting and metallic phases is dramatically enhanced in the visible region, presenting size-dependent optical resonances and size-dependent transition temperatures. The collective optical response as a function of temperature presents an enhanced scattering state during the evolving phase transition. The effects appear to arise because of the underlying VO2 mesoscale optical properties, the heterogeneous nucleation behind the phase transition, and the incoherent coupling between the nanoparticles undergoing an order-disorder-order transition. Calculations that support these interpretations are presented. PMID- 15525127 TI - Do spherical polyelectrolyte brushes interdigitate? AB - The structure of spherical micelles of the diblock copolymer poly(styrene-block acrylic acid) in water was investigated with small angle neutron scattering and contrast matching. We have monitored intermicelle correlation and the extension of the polyelectrolyte chains in the coronal layer through the overlap concentration. Irrespective of ionic strength, the corona shrinks with increasing packing fraction. Furthermore, at high charge and minimal screening conditions, the corona layers interpenetrate once the volume fraction exceeds the critical value 0.53+/-0.02. PMID- 15525128 TI - Enhancement of association rates by nonspecific binding to DNA and cell membranes. AB - A comprehensive analytic theory is developed for the kinetics of reversible association with specific sites on DNA and receptors on cell membranes in the presence of nonspecific binding to the target surfaces. Nonspecific binding is treated as a short-range attractive potential, which is more fundamental and realistic than the surface sliding model. The presence of a surface potential around the target enhances the rate of specific association and for reversible reactions leads to deviations from single exponential relaxation. PMID- 15525129 TI - Species abundance patterns in complex evolutionary dynamics. AB - An analytic theory of species abundance patterns (SAPs) in biological networks is presented. The theory is based on multispecies replicator dynamics equivalent to the Lotka-Volterra equation, with diverse interspecies interactions. Various SAPs observed in nature are derived from a single parameter. The abundance distribution is formed like a widely observed left-skewed lognormal distribution. As the model has a general form, the result can be applied to similar patterns in other complex biological networks, e.g., gene expression. PMID- 15525130 TI - Critical scale invariance in a healthy human heart rate. AB - We demonstrate the robust scale-invariance in the probability density function (PDF) of detrended healthy human heart rate increments, which is preserved not only in a quiescent condition, but also in a dynamic state where the mean level of the heart rate is dramatically changing. This scale-independent and fractal structure is markedly different from the scale-dependent PDF evolution observed in a turbulentlike, cascade heart rate model. These results strongly support the view that a healthy human heart rate is controlled to converge continually to a critical state. PMID- 15525131 TI - Self-diffusion in sheared colloidal suspensions: violation of fluctuation dissipation relation. AB - Using memory-function formalism we show that in sheared colloidal suspensions the fluctuation-dissipation theorem for self-diffusion, i.e., Einstein's relation between self-diffusion and mobility tensors, is violated and propose a new way to measure this violation in Brownian dynamics simulations. We derive mode-coupling expressions for the tagged particle friction tensor and for an effective, shear rate dependent temperature. PMID- 15525132 TI - Strong temperature dependence of irradiation effects in organic layers. AB - Radiation damage of self-assembled monolayers, which are prototypes of thin organic layers and highly organized biological systems, shows a strong dependence on temperature. Two limiting cases could be identified. Reactions involving transport of single atoms and small fragments proceed nearly independent of temperature. Reactions requiring transport of heavy fragments are, however, efficiently quenched by cooling. We foresee the combined use of temperature and irradiation by electrons or photons for advanced tailoring of self-assembled monolayers on surfaces. In addition, our results have direct implications for cryogenic approaches in advanced electron and x-ray microscopy and spectroscopy of biological macromolecules and cells. PMID- 15525133 TI - Scale dependence and localization of the deformation of Arctic sea ice. AB - A scaling analysis of the deformation of Arctic sea ice over a 3-day time period is performed for scales of 10 to 1000 km. The deformation field is derived from satellite radar data; it allows us to study how a very large solid body-the Arctic sea-ice cover-deforms under the action of heterogeneous forcing winds and ocean currents. The deformation is strongly localized at small scales, and can be characterized as multifractal. This behavior is well known for turbulent flows, and is here also observed for a deforming solid. A multiscaling extrapolation to the meter scale (laboratory scale) shows that, at the 3-day time scale, about 15% of the deformation is larger than 10(-4) s(-1), implying brittle failure, over 0.2% of the total area. PMID- 15525134 TI - Observation of quantum oscillations between a Josephson phase qubit and a microscopic resonator using fast readout. AB - We have detected coherent quantum oscillations between Josephson phase qubits and critical-current fluctuators by implementing a new state readout technique that is an order of magnitude faster than previous methods. These results reveal a new aspect of the quantum behavior of Josephson junctions, and they demonstrate the means to measure two-qubit interactions in the time domain. The junction fluctuator interaction also points to a possible mechanism for decoherence and reduced fidelity in superconducting qubits. PMID- 15525135 TI - Limit to minimally destructive optical detection of atoms. AB - All optical techniques used to probe the properties of Bose-Einstein condensates have been based on dispersion and absorption that can be described by a two-level atom. Both phenomena lead to spontaneous emission that is destructive at the low energies involved with ultracold atomic systems. Recently, both were shown to lead to the same limit to the signal to noise ratio for a given destruction. We develop a new method for calculating the phase shift of a laser beam and show that no single-pass optical technique using classical light and a three-level atom can exceed this limit. This puts significant restrictions on potential nondestructive measurement schemes. PMID- 15525136 TI - Interference of an array of independent Bose-Einstein condensates. AB - We have observed high-contrast matter wave interference between 30 Bose-Einstein condensates with uncorrelated phases. Interferences were observed after the independent condensates were released from a one-dimensional optical lattice and allowed to overlap. This phenomenon is explained with a simple theoretical model, which generalizes the analysis of the interference of two condensates. PMID- 15525137 TI - How quantum correlations enhance prediction of complementary measurements. AB - If there are correlations between two qubits, then the results of the measurement on one of them can help to predict measurement results on the other one. It is an interesting question as to what can be predicted about the results of two complementary projective measurements on the first qubit. To quantify these predictions the complementary knowledge excesses are used. A nontrivial constraint restricting them is derived. For any mixed state and for arbitrary measurements the knowledge excesses are bounded by a factor depending only on the maximal violation of Bell's inequalities. This result is experimentally verified on two-photon Werner states prepared by means of spontaneous parametric down conversion. PMID- 15525138 TI - Strong Levinson theorem for the Dirac equation. AB - We consider the Dirac equation in one space dimension in the presence of a symmetric potential well. We connect the scattering phase shifts at E= +m and E= m to the number of states that have left the positive energy continuum or joined the negative energy continuum, respectively, as the potential is turned on from zero. PMID- 15525139 TI - Confined quantum time of arrivals. AB - We show that formulating the quantum time of arrival problem in a segment of the real line suggests rephrasing the quantum time of arrival problem to finding states that evolve to unitarily collapse at a given point at a definite time. For the spatially confined particle, we show that the problem admits a solution in the form of an eigenvalue problem of a compact and self-adjoint time of arrival operator derived by a quantization of the classical time of arrival, which is canonically conjugate with the Hamiltonian in a closed subspace of the Hilbert space. PMID- 15525140 TI - Electron-positron pair production by photons in nonuniform strong fields. AB - A probability of electron-positron pair production by photons in strong nonuniform fields is derived by applying a model trajectory method in the frame of a semiclassical approach. In addition to the well known invariant field parameter chi, a new invariant parameter nu is introduced to characterize the nonuniformity of the field. For nu>>1, the obtained expression is reduced to the uniform-field approximation while it approaches the Bethe-Heitler formula for nu<<1. The pair production is predicted for relatively weak external fields where the uniform-field approximation gives no effect. The theory agrees well with the experimental results of crystal-assisted pair production. PMID- 15525141 TI - Quantum cloning of a coherent light state into an atomic quantum memory. AB - A scheme for the optimal Gaussian cloning of coherent light states at the interface between light and atoms is proposed. The distinct feature of this proposal is that the clones are stored in an atomic quantum memory, which is important for applications in quantum communication. The atomic quantum cloning machine requires only a single passage of the light pulse through the atomic ensembles followed by the measurement of a light quadrature and an appropriate feedback, which renders the protocol experimentally feasible. An alternative protocol, where one of the clones is carried by the outgoing light pulse, is discussed in connection with eavesdropping on quantum key distribution. PMID- 15525142 TI - Distribution of time-bin entangled qubits over 50 km of optical fiber. AB - We report experimental distribution of time-bin entangled qubits over 50 km of optical fibers. Using actively stabilized preparation and measurement devices we demonstrate violation of the Clauser-Horne-Shimony-Holt Bell inequality by more than 15 standard deviations without removing the detector noise. In addition we report a proof-of principle experiment of quantum key distribution over 50 km of optical fibers using entangled photon. PMID- 15525143 TI - Efficient use of quantum resources for the transmission of a reference frame. AB - We propose a covariant protocol for transmitting reference frames encoded on N spins, achieving sensitivity N-2 without the need of a preestablished reference frame and without using entanglement between sender and receiver. The protocol exploits the use of equivalent representations that were overlooked in the previous literature. PMID- 15525144 TI - Quasiperiodic dynamics of a quantum walk on the line. AB - We study the dynamics of a generalization of a quantum coin walk on the line, which is a natural model for a diffusion modified by quantum or interference effects. In particular, our results provide surprisingly simple explanations for recurrence phenomena observed by Bouwmeester et al. [Phys. Rev. A 61, 13410 (1999)]] in their optical Galton board experiment, and a description of a stroboscopic quantum walk given by Buerschaper and Burnett [quant-ph/0406039] through numerical simulations. We also provide heuristic explanations for the behavior of our model which show, in particular, that its dynamics can be viewed as a discrete version of Bloch oscillations. PMID- 15525145 TI - Stochastic reaction-diffusion phenomena in porous media with nonlinear kinetics: effects of quenched porosity fluctuations. AB - We study the effects of quenched porosity fluctuations in the presence of nonlinear kinetics in a reaction-diffusion porous system. Adopting a weak-noise approximation and a mean-field assumption, a renormalized equation of motion for the average concentration of a chemical species is obtained. It is characterized by a smaller effective diffusion coefficient and by the presence of supplementary nonlinear reactive terms. Explicit results are given for a Gaussian porosity distribution and for a long-ranged one. Comparisons with simulations in one dimension are presented. PMID- 15525146 TI - Generalized Langevin equation with fractional Gaussian noise: subdiffusion within a single protein molecule. AB - By introducing fractional Gaussian noise into the generalized Langevin equation, the subdiffusion of a particle can be described as a stationary Gaussian process with analytical tractability. This model is capable of explaining the equilibrium fluctuation of the distance between an electron transfer donor and acceptor pair within a protein that spans a broad range of time scales, and is in excellent agreement with a single-molecule experiment. PMID- 15525147 TI - Tip-enhanced fluorescence microscopy at 10 nanometer resolution. AB - We demonstrate unambiguously that the field enhancement near the apex of a laser illuminated silicon tip decays according to a power law that is moderated by a single parameter characterizing the tip sharpness. Oscillating the probe in intermittent contact with a semiconductor nanocrystal strongly modulates the fluorescence excitation rate, providing robust optical contrast and enabling excellent background rejection. Laterally encoded demodulation yields images with <10 nm spatial resolution, consistent with independent measurements of tip sharpness. PMID- 15525149 TI - Phantom energy mediates a long-range repulsive force. AB - Scalar field models with nonstandard kinetic terms have been proposed in the context of k inflation, of Born-Infeld Lagrangians, of phantom energy and, more in general, of low-energy string theory. In general, scalar fields are expected to couple to matter inducing a new interaction. In this Letter I derive the cosmological perturbation equations and the Yukawa correction to gravity for such general models. I find three interesting results: first, when the field behaves as phantom energy (equation of state less than -1), then the coupling strength is negative, inducing a long-range repulsive force; second, the dark-energy field might cluster on astrophysical scales; third, applying the formalism to a Brans Dicke theory with a general kinetic term it is shown that its Newtonian effects depend on a single parameter that generalizes the Brans-Dicke constant. PMID- 15525148 TI - TeV neutrinos from core collapse supernovae and hypernovae. AB - A fraction of core-collapse supernovae of type Ib/c are associated with gamma-ray bursts, which are thought to produce highly relativistic jets. Recently, it has been hypothesized that a larger fraction of core-collapse supernovae produce slower jets, which may contribute to the disruption and ejection of the supernova envelope, and explain the unusually energetic hypernovae. We explore the TeV neutrino signatures expected from such slower jets, and calculate the expected detection rates with upcoming Gigaton Cherenkov experiments. We conclude that individual jetted supernovae may be detectable from nearby galaxies. PMID- 15525151 TI - A determination of the Cabibbo-Kobayashi-Maskawa parameter |V us| using KL decays. AB - We present a determination of the Cabibbo-Kobayashi-Maskawa parameter |V(us)| based on new measurements of the six largest K(L) branching fractions and semileptonic form factors by the KTeV (E832) experiment at Fermilab. We find |V(us)|=0.2252+/-0.0008(KTeV)+/-0.0021(ext), where the errors are from KTeV measurements and from external sources. We also use the measured branching fractions to determine the CP violation parameter |eta(+-)|=(2.228+/ 0.005(KTeV)+/-0.009(ext))x10(-3). PMID- 15525152 TI - Precise measurement of the pi+-->pi0 e+nu branching ratio. AB - Using a large acceptance calorimeter and a stopped pion beam we have made a precise measurement of the rare pi(+)-->pi(0)e(+)nu (pi(beta)) decay branching ratio. We have evaluated the branching ratio by normalizing the number of observed pi(beta) decays to the number of observed pi(+)-->e(+)nu (pi(e2)) decays. We find the value of Gamma(pi(+)-->pi(0)e(+)nu)/Gamma(total)=[1.036+/ 0.004(stat)+/-0.004(syst)+/-0.003(pi(e2))]x10(-8), where the first uncertainty is statistical, the second systematic, and the third is the pi(e2) branching ratio uncertainty. Our result agrees well with the standard model prediction. PMID- 15525153 TI - Precise measurement of the pion axial form factor in the pi+-->e+nugamma decay. AB - We have studied radiative pion decays pi(+)-->e(+)nugamma in three broad kinematic regions using the PIBETA detector and a stopped pion beam. Based on Dalitz distributions of 41 601 events we have evaluated absolute pi-->enugamma branching ratios in the three regions. Minimum chi(2) fits to the integral and differential (E(e(+)),E(gamma)) distributions result in the axial-to-vector weak form factor ratio of gamma identical with F(A)/F(V)=0.443(15), or F(A)=0.0115(4) with F(V)=0.0259. However, deviations from standard model predictions in the high E(gamma)-low-E(e(+)) kinematic region indicate the need for further theoretical and experimental work. PMID- 15525156 TI - Fluctuations of the initial conditions and the continuous emission in the hydrodynamical description of two-pion interferometry. AB - Within the hydrodynamical approach, we study the Bose-Einstein correlation of identical pions by taking into account both event-by-event fluctuating initial conditions and continuous pion emission during the whole development of the hot and dense matter formed in high-energy collisions. Important deviations occur, compared to the usual hydrocalculations with smooth initial conditions and a sudden freeze-out on a well defined hypersurface. Comparison with data at the BNL Relativistic Heavy Ion Collider (RHIC) shows that, despite the rather rough approximation we used here, this description can account for the m(T) dependence of R(L) and R(s), and produces a significant improvement for R(o) with respect to the usual version. PMID- 15525157 TI - Record of cycling operation of the natural nuclear reactor in the Oklo/Okelobondo area in Gabon. AB - Using selective laser extraction technique combined with sensitive ion-counting mass spectrometry, we have analyzed the isotopic structure of fission noble gases in U-free La-Ce-Sr-Ca aluminous hydroxy phosphate associated with the 2 billion yr old Oklo natural nuclear reactor. In addition to elevated abundances of fission-produced Zr, Ce, and Sr, we discovered high (up to 0.03 cm(3) STP/g) concentrations of fission Xe and Kr, the largest ever observed in any natural material. The specific isotopic structure of xenon in this mineral defines a cycling operation for the reactor with 30-min active pulses separated by 2.5 h dormant periods. Thus, nature not only created conditions for self-sustained nuclear chain reactions, but also provided clues on how to retain nuclear wastes, including fission Xe and Kr, and prevent uncontrolled runaway chain reaction. PMID- 15525159 TI - Relativistic and interchannel coupling effects in photoionization angular distributions by synchrotron spectrocopy of laser cooled atoms. AB - We investigate the angular distribution of photoionization fragments at low photon energies (12-40 eV) in an open shell atom, by synchrotron radiation recoil ion momentum spectroscopy in a laser cooled and trapped sample. For cesium atoms, for which relativistic effects play an important role and the ion recoil is relatively small, we could determine large and rapid changes of the asymmetry parameter beta from two, observed for s electrons outside resonances and far from the Cooper minimum. They can be explained by relativistic effects and interchannel coupling arising from final state configuration mixing. PMID- 15525158 TI - Correlated strength in the nuclear spectral function. AB - We have carried out an (e,e'p) experiment at high momentum transfer and in parallel kinematics to measure the strength of the nuclear spectral function S(k,E) at high nucleon momenta k and large removal energies E. This strength is related to the presence of short-range and tensor correlations, and was known hitherto only indirectly and with considerable uncertainty from the lack of strength in the independent-particle region. This experiment locates by direct measurement the correlated strength predicted by theory. PMID- 15525160 TI - Observation of heteronuclear Feshbach resonances in a mixture of bosons and fermions. AB - Three magnetic-field induced heteronuclear Feshbach resonances were identified in collisions between bosonic 87Rb and fermionic 40K atoms in their absolute ground states. Strong inelastic loss from an optically trapped mixture was observed at the resonance positions of 492, 512, and 543+/-2 G. The magnetic-field locations of these resonances place a tight constraint on the triplet and singlet cross species scattering lengths, yielding (-281+/-15)a(0) and (-54+/-12)a(0), respectively. The width of the loss feature at 543 G is 3.7+/-1.5 G wide; this broad Feshbach resonance should enable experimental control of the interspecies interactions. PMID- 15525161 TI - Routes to control of H2 Coulomb explosion in few-cycle laser pulses. AB - We have measured coincident ion pairs produced in the Coulomb explosion of H2 by 8-30 fs laser pulses at different laser intensities. We show how the Coulomb explosion of H2 can be experimentally controlled by tuning the appropriate pulse duration and laser intensity. For laser pulses less than 15 fs, we found that the rescattering-induced Coulomb explosion is dominated by first-return recollisions, while for longer pulses and at the proper laser intensity, the third return can be made to be the major one. Additionally, by choosing suitable pulse duration and laser intensity, we show H2 Coulomb explosion proceeding through three distinct processes that are simultaneously observable, each exhibiting different characteristics and revealing distinctive time information about the H2 evolution in the laser pulse. PMID- 15525162 TI - Regge oscillations in integral cross sections for proton impact on atomic hydrogen. AB - The integral cross sections for elastic scattering and spin exchange for proton impact on atomic hydrogen show several oscillations in the energy range 0.01-1.0 eV that cannot be associated with resonances or the glory effect. A complex angular momentum analysis using computed Regge trajectories shows that each peak of the oscillatory structure is predominantly associated with at most three trajectories. In this way, the peaks are related to the L=0 bound states of H+ 2. The complex angular momentum theory for integral cross sections that we introduce shows that such oscillations are a general feature of potential scattering. PMID- 15525163 TI - Frequency mixing using electromagnetically induced transparency in cold atoms. AB - We report the first experimental demonstration of four-wave mixing using electromagnetically induced transparency in cold atoms. Backward-wave, phase matched difference-frequency conversion is achieved at optical powers of a few nanowatts and at energies of less than a picojoule. PMID- 15525164 TI - Vector soliton fission. AB - We investigate the vectorial nature of soliton fission in an isotropic nonlinear medium both theoretically and experimentally. As a specific example, we show that supercontinuum generation in a tapered fiber is extremely sensitive to the input state of polarization. Multiple vector solitons generated through soliton fission exhibit different states of elliptical polarization while emitting nonsolitonic radiation with complicated polarization features. Experiments performed with a tapered fiber agree with our theoretical description. PMID- 15525165 TI - Thermal diode: rectification of heat flux. AB - By coupling two nonlinear one dimensional lattices, we demonstrate a thermal diode model that works in a wide range of system parameters. We provide numerical and analytical evidence for the underlying mechanism which allows heat flux in one direction while the system acts like an insulator when the temperature gradient is reversed. The possible experimental realization in nanoscale systems is briefly discussed. PMID- 15525166 TI - Stripe formation in granular mixtures due to the differential influence of drag. AB - We describe an investigation of fluid-immersed granular mixtures subjected to horizontal vibration. For sufficiently large amplitudes of vibration, a mixture of equal-sized glass and bronze particles in water is found to separate into a striped pattern. Numerical simulations based on soft-sphere molecular dynamics coupled to the interstitial fluid are able to capture many of the features observed experimentally. We propose a general pattern-formation mechanism based on the differential influence of drag on the components of the mixture. An expression for the number of stripes as a function of the system parameters is derived and shown to be in good agreement with experiments. PMID- 15525167 TI - Pattern formation in binary fluids confined between rough, chemically heterogeneous surfaces. AB - Using a mesoscale model for hydrodynamics, we simulate driven flow of AB binary fluids past surfaces that contain well-defined roughness or asperities. The geometry and wetting properties of the asperities are found to have a dramatic effect on the flow patterns. We isolate conditions where the A fluid forms vertical bands that bridge the asperities and an imposed shear (or pressure gradient) drives the system to form monodisperse droplets of A within the B fluid. The size of the droplets can be tailored by varying the morphology of the asperities. The surfaces needed to create this rich dynamical behavior are used as the stamps in microcontact printing; thus, the parameter space can readily be accessed experimentally, and the predictions suggest an efficient method for forming emulsions with well-controlled morphologies. PMID- 15525168 TI - Viscous entrainment from a nozzle: singular liquid spouts. AB - We analyze a long-wavelength model of viscous entrainment from a nozzle and show that, when appropriate large-scale boundary conditions are imposed, the entrainment transition can become continuous. As the entrainment threshold is approached from above, the entrained steady-state spout becomes vanishingly thin compared to the nozzle radius and takes the form of a thin tendril emerging from a base profile whose tip is perfectly conical. PMID- 15525169 TI - Velocity and velocity-difference distributions in Burgers turbulence. AB - We consider the one-dimensional Burgers equation randomly stirred at large scales by a Gaussian short-time correlated force. Using the method of dissipative anomalies, we obtain velocity and velocity-difference probability density functions and confirm the results with high-resolution numerical simulations. PMID- 15525170 TI - Generation of 0.4-keV femtosecond electron pulses using impulsively excited surface plasmons. AB - We demonstrate the generation of 0.4-keV, sub-27 fs electron pulses using low intensity laser pulses from a Ti:sapphire oscillator through the excitation of surface plasmon waves on a time scale within the plasmon lifetime. Modeling of the ponderomotive electron pulse acceleration yields electron energy spectra that are in excellent agreement with the observed ones. Our work opens a doorway for time-resolved experimentation using low-power, high-repetition rate laser pulses. PMID- 15525171 TI - Diagnostics of the electronegative plasma sheath at low pressures using microparticles. AB - Levitated particles are a new powerful diagnostic of the midplasma sheath region. They can reveal features undetectable either to plasma or to surface measurements. The equilibrium position of microparticles suspended in an oxygen plasma sheath, together with a model of the levitation force and Langmuir probe measurements, gives evidence of secondary electropositive plasmas in the already established plasma sheath, in the range of parameters where the modified Bohm criterion breaks down into multiple solutions. PMID- 15525172 TI - Near-GeV-energy laser-wakefield acceleration of self-injected electrons in a centimeter-scale plasma channel. AB - The first three-dimensional, particle-in-cell (PIC) simulations of laser wakefield acceleration of self-injected electrons in a 0.84 cm long plasma channel are reported. The frequency evolution of the initially 50 fs (FWHM) long laser pulse by photon interaction with the wake followed by plasma dispersion enhances the wake which eventually leads to self-injection of electrons from the channel wall. This first bunch of electrons remains spatially highly localized. Its phase space rotation due to slippage with respect to the wake leads to a monoenergetic bunch of electrons with a central energy of 0.26 GeV after 0.55 cm propagation. At later times, spatial bunching of the laser enhances the acceleration of a second bunch of electrons to energies up to 0.84 GeV before the laser pulse intensity is significantly reduced. PMID- 15525155 TI - Searches for B0 decays to combinations of two charmless isoscalar mesons. AB - We search for B meson decays into two-body combinations of eta, eta', omega, and phi mesons from 89 x 10(6) BB pairs collected with the BABAR detector at the PEP II asymmetric-energy e+e- collider at SLAC. We find the branching fraction B(B0- >etaomega)=(4.0(+1.3)(-1.2)+/-0.4)x10(-6) with a significance of 4.3 sigma. For the other decay modes we set the following 90% confidence level upper limits on the branching fractions, in units of 10(-6): B(B0-->etaeta)<2.8, B(B0- >etaeta')<4.6, B(B0-->eta'eta')<10, B(B0-->eta'omega)<2.8, B(B0-->etaphi)<1.0, B(B0-->eta'phi)<4.5, and B(B0-->phiphi)<1.5. PMID- 15525173 TI - Breakdown of hot-spot model in determining convective amplification in large homogeneous systems. AB - Convective amplification in large homogeneous systems is studied, both analytically and numerically, in the case of a linear diffraction-free stochastic amplifier. Overall amplification does not result from successive amplifications in small scale high intensity hot spots, but from a single amplification in a delocalized mode of the driver field spreading over the whole interaction length. For this model, the hot-spot approach is found to systematically underestimate the gain factor by more than 50%. PMID- 15525150 TI - Study of B-->D(*)sJ +-D(*) decays. AB - We report a study of D(*)(sJ)(2317)(+) and D(sJ)(2460)(+) meson production in B decays. We observe the decays B+-->D((*)+)(sJ)D ((*)0) and B0-->D((*)+)(sJ)D((*) ) with the subsequent decays D(*)(sJ)(2317)(+)-->D(+)(s)pi(0), D(sJ)(2460)(+)- >D(+)(s)gamma, and D(sJ)(2460)(+)-->D(*+)(s)pi(0). Based on a data sample of 122.1 x 10(6) BB pairs collected with the BABAR detector at the PEP-II B factory, we obtain branching fractions for these modes, including the previously unseen decays B-->D((*)+)(sJ)D(*). In addition, we perform an angular analysis of D(sJ)(2460)(+)-->D(+)(s)gamma decays to test the different D(sJ)(2460)(+) spin hypotheses. PMID- 15525174 TI - Global simulation for laser-driven MeV electrons in fast ignition. AB - A comprehensive examination of the interaction of a picosecond-long ignition pulse on high-density (40 times critical density) pellets using a two-dimensional particle-in-cell model is described. The global geometry consists of a 50 mum diameter pellet surrounded by a corona which is isolated by a vacuum region from the boundary. For cone-attached targets, as much as 67% of the incident laser energy is absorbed with 12% sent forward as fast electrons in a 23 degrees cone. The current filaments are driven by the Weibel instability of the forward-going fast electron flux and its return current with the ions playing an important role of neutralizing the space charge. No global current filament coalescence has been observed. The electron distribution function obeys a power law, which begins at E approximately 0.2 MeV and falls off as E-(2-3). PMID- 15525175 TI - Intrinsic energy loss mechanisms in a cantilevered carbon nanotube beam oscillator. AB - Classical molecular dynamics is applied to study the energy dissipation (the Q factor) of the cantilever-type beam oscillators of single wall and double-walled carbon nanotubes (CNTs). The study finds that the Q factor of the CNT beam oscillator varies with the temperature T following the 1/T(0.36) dependence. For single wall CNT, the Q factor drops from 2 x 10(5) at 0.05 K to 1.5 x 10(3) at 293 K. The study further reveals that the weak interlayer binding strength and the interlayer commensurance significantly increases the energy dissipation in the double-walled CNT oscillator. PMID- 15525176 TI - Path prediction of kinked and branched cracks in plane situations. AB - Using the asymptotic expansion of the stress field ahead a curved extension of a straight crack, some general results on the paths selected by kinked and branched cracks are derived. When dealing with the dynamic branching instability of a single propagation crack, the experimentally observed shape of the branches is recovered without introducing any adjustable parameter. It is shown that the length scale introduced by the curved extension of the branches is given by the geometrical length scale of the experiment. The theoretical results agree quantitatively with the experimental findings. PMID- 15525177 TI - Kohn anomalies and electron-phonon interactions in graphite. AB - We demonstrate that graphite phonon dispersions have two Kohn anomalies at the Gamma-E(2g) and K-A'1 modes. The anomalies are revealed by two sharp kinks. By an exact analytic derivation, we show that the slope of these kinks is proportional to the square of the electron-phonon coupling (EPC). Thus, we can directly measure the EPC from the experimental dispersions. The Gamma-E(2g) and K-A'1 EPCs are particularly large, while they are negligible for all the other modes at Gamma and K. PMID- 15525154 TI - Branching fractions and CP asymmetries in B0-->K+ K- K0 S and B+-->K+ K0 S K0 S. AB - We measure the branching fractions and CP asymmetries in the decays B0-->K(+)K( )K(0)(S) and B+-->K(+)K(0)(S)K(0)(S) using a sample of approximately 122x10(6) BB pairs collected by the BABAR detector. From a time-dependent analysis of the K(+)K(-)K(0)(S) sample that excludes phiK(0)(S), the values of the CP-violation parameters are S=-0.56+/-0.25+/-0.04 and C=-0.10+/-0.19+/-0.10, where the first uncertainty is statistical and the second is systematic. We confirm that the final state is nearly purely CP even and extract the standard model parameter sin(2beta=0.57+/-0.26+/-0.04(+0.17)(-0) where the last error is due to uncertainty on the CP content. We present the first measurement of the CP violating charge asymmetry A(CP)(B+-->K(+)K(0)(S)K(0)(S))=-0.04+/-0.11+/-0.02. The branching fractions are B(B0-->K+K-K0)=(23.8+/-2.0+/-1.6)x10(-6) and B(B+- >K(+)K(0)(S)K(0)(S))=(10.7+/-1.2+/-1.0)x10(-6). PMID- 15525178 TI - Peculiar effects of anisotropic diffusion on dynamics of vicinal surfaces. AB - We report on peculiar behaviors due to anisotropic terrace diffusion on step meandering on a vicinal surface. We find that anisotropy triggers tilted ripples. In addition, if the fast diffusion direction is perpendicular to the steps, the instability is moderate and coarsening is absent, while in the opposite case the instability is promoted, and interrupted coarsening may be observed. Strong enough anisotropy restabilizes the step for almost all step orientations. These findings point to the nontrivial effect of anisotropy and open promising lines of inquiries in the design of surface architectures. PMID- 15525180 TI - Disorder by random crosslinking in smectic elastomers. AB - We present a high-resolution x-ray study of the effects of disorder due to random crosslinking on the one-dimensional translational ordering in smectic elastomers. At a small crosslink density of about 5%, the elastomer network stabilizes the smectic structure against layer-displacement fluctuations, and the algebraically decaying layer ordering extends up to several micrometers. With increasing concentration of crosslinks, the finite size of these domains is strongly reduced, indicating that disordering takes over. Finally, at a crosslink concentration of 20%, the structure factor can be described by a Lorentzian, which signals extended short-range correlations. The findings are discussed in terms of recent theories of randomly quenched disorder. PMID- 15525179 TI - Nanoscale hydrophobic interaction and nanobubble nucleation. AB - We report large-scale atomistic simulation of midrange nanoscale hydrophobic interaction, manifested by the nucleation of nanobubble between nanometer-sized hydrophobes at constrained equilibrium. When the length scale of the hydrophobes is greater than 2 nm, the nanobubble formation shows hysteresis behavior resembling the first-order transition. Calculation of the potential of mean force versus interhydrophobe distance provides a quantitative measure of the strength of the nanoscale hydrophobic interaction. PMID- 15525181 TI - Investigations of the structure of water using mid-IR fiberoptic evanescent wave spectroscopy. AB - Liquid water is one of the most studied but still one of the least understood substances. The absorption spectra of water in the mid-IR were measured from -10 and up to 90 degrees C using fiberoptic evanescent wave spectroscopy. The changes in the spectrum and the existence of an isosbestic point during the ice-liquid phase transition were studied. Based on the spectroscopic data we propose a multispecies model for the structure of liquid water. The new model provides an explanation for the results of the measurements reported in this work. It also provides an explanation for some of the unique thermodynamic and kinetic properties of liquid water and for many of the anomalies of water. PMID- 15525182 TI - Negative entropy of mixing for vanadium-platinum solutions. AB - The phonon densities of states for pure vanadium and the solid solutions V-6.25% Ni, Pd, Pt were determined from inelastic neutron scattering measurements. The solute atoms caused a large stiffening of the phonons, resulting in large, negative vibrational entropies of mixing. For V-6.25%Pt, the negative vibrational entropy of mixing exceeds the conventional positive chemical entropy of mixing. This negative total entropy of mixing should extend to lower concentrations of Pt, and the effect on the bcc solvus line is discussed. The experimental data were inverted to obtain interatomic force constants by using a Born-von Karman model with an iterative optimization algorithm. The stiffening of bonds responsible for the decrease of entropy was found to occur mainly in first nearest-neighbor solute-host bonds, and correlates in part with the solute metallic radius. PMID- 15525183 TI - Optimal swimming at low Reynolds numbers. AB - Efficient swimming at low Reynolds numbers is a major concern of microbots. To compare the efficiencies of different swimmers we introduce the notion of "a swimming drag coefficient" which allows for the ranking of swimmers. We find the optimal swimmer within a certain class of two-dimensional swimmers using conformal mapping techniques. PMID- 15525184 TI - Fractional statistical theory of adsorption of polyatomics. AB - A new theoretical description of fractional statistical theory of adsorption (FSTA) phenomena is presented based on Haldane's statistics. Thermodynamic functions for adsorption of polyatomics are analytically developed. The entropy is characterized by an exclusion parameter g, which relates to the configuration of the admolecules and surface geometry. FSTA provides a simple framework to address a large class of complex adsorption systems. Comparisons of theoretical adsorption isotherms with experiments and simulations indicate that adsorption configuration and adsorption energy can accurately be assessed from this theory. PMID- 15525185 TI - Magnetic properties of (BEDT-TTF)(ClMeTCNQ) studied by ESR under pressure: a neutral-ionic crossover and thermoinduced mesophase. AB - The neutral-ionic (NI) phase transition in (BEDT-TTF)(ClMeTCNQ) is studied with ESR under pressure to tune the transition temperature. Broad peak structures observed in the spin susceptibility around T(NI) are analyzed with the assumption of a spin-Peierls gap of 1100 K, which demonstrates the continuous evolution of ionic domain fraction through T(NI) over the extent T(W) proportional, variantT(NI). An unexpected feature is the presence of sizable ionic droplets even at T>>T(NI), which is ascribed to the tiny energy difference between the two phases. PMID- 15525186 TI - Well-defined quasiparticles in interacting metallic grains. AB - We analyze spectral functions of mesoscopic systems with large dimensionless conductance, which can be described by a universal Hamiltonian. We show that an important class of spectral functions are dominated by one single state only, which implies the existence of well-defined (i.e., infinite-lifetime) quasiparticles. Furthermore, the dominance of a single state enables us to calculate zero-temperature spectral functions with high accuracy using the density-matrix renormalization group. We illustrate the use of this method by calculating the tunneling density of states of metallic grains, of which we discuss the crossover from the few-electron to the bulk regime. PMID- 15525187 TI - Jahn-Teller distortions and the supershell effect in metal nanowires. AB - A stability analysis of metal nanowires shows that a Jahn-Teller deformation breaking cylindrical symmetry can be energetically favorable, leading to stable nanowires with elliptic cross sections. The sequence of stable cylindrical and elliptical nanowires allows for a consistent interpretation of experimental conductance histograms for alkali metals, including both the electronic shell and supershell structures. It is predicted that for gold, elliptical nanowires are even more likely to form since their eccentricity is smaller than for alkali metals. The existence of certain metastable superdeformed nanowires is also predicted. PMID- 15525188 TI - Strong exciton-photon coupling and exciton hybridization in a thermally evaporated polycrystalline film of an organic small molecule. AB - We demonstrate strong exciton-photon coupling in an optical microcavity containing a thermally evaporated polycrystalline organic thin film. Microcavity polaritons result from coupling between the 0-0 excitonic transition of 3,4,7,8 napthalenetetracarboxylic dianhydride and a cavity photon. For thicker films, the 0-1 transition also couples to the cavity mode, as vibronic relaxation is overcome by the short Rabi period for strong coupling. To our knowledge, this is the first report of strong coupling between a cavity photon and multiple vibronic transitions in a single material, made possible by the pronounced vibronic absorption features characteristic of crystalline organic materials. PMID- 15525189 TI - 4f-electron localization in CexLa 1-xM In5 with M=Co, Rh, or Ir. AB - de Haas-van Alphen measurements on Ce(x)La(1-x)MIn(5) yield contrasting types of behavior that depend on whether M=Co and Ir or M=Rh. A stronger x-dependent scattering in the case of M=Co and Ir is suggestive of a stronger relative coupling, J/W, of the conduction electrons to the 4f electrons, which would then account for the development of a heavy composite Fermi-liquid state as x-->1. The failure of a composite Fermi-liquid state to form for any x in the case of M= Rh is shown to be inconsistent with theoretical models that propose antiferromagnetism to result from spin-density-wave formation. PMID- 15525190 TI - Periodic field emission from an isolated nanoscale electron island. AB - We observe field emission from an isolated nanomachined gold island. The island is able to mechanically oscillate between two facing electrodes, which provide recharging and detection of the emission current. We are able to trace and reproduce the transition from current flow through a rectangular tunneling barrier to the regime of field emission. A theoretical model via a master equation reproduces the experimental data and shows deviation from the Fowler Nordheim description due to the island's electric isolation. PMID- 15525191 TI - Manipulation of a single charge in a double quantum dot. AB - We manipulate a single electron in a fully tunable double quantum dot using microwave excitation. Under resonant conditions, microwaves drive transitions between the (1,0) and (0,1) charge states of the double dot. Local quantum point contact charge detectors enable a direct measurement of the photon-induced change in occupancy of the charge states. From charge sensing measurements, we find T1 approximately 16 ns and a lower bound estimate for T*(2) of 400 ps for the charge two-level system. PMID- 15525192 TI - Electronic structure and bonding of Au on a SiO2 cluster: a nanobullet for tumors. AB - The binding of gold atoms to a small silica cluster has been studied by using density functional theory with generalized gradient approximation for exchange and correlation. It is shown that gold atoms bind to silicon atoms with dangling bonds and serve as seeds for the growth of Au islands. The large electron affinity of gold causes a significant change in the electronic structure of silica resulting in a substantial reduction in the highest occupied and the lowest unoccupied molecular orbital and the optical gap, thus allowing it to absorb near infrared radiation. This suggests that a small cluster can have a similar functionality in the treatment of cancer as the large size nanoshell, but for a different mechanism. PMID- 15525193 TI - Quantized dispersion of two-dimensional magnetoplasmons detected by photoconductivity spectroscopy. AB - We find that the long-wavelength magnetoplasmon, resistively detected by photoconductivity spectroscopy in high-mobility two-dimensional electron systems, deviates from its well-known semiclassical nature as uncovered in conventional absorption experiments. A clear filling-factor dependent plateau-type dispersion is observed that reveals a so far unknown relation between the magnetoplasmon and the quantum Hall effect. PMID- 15525194 TI - Fano versus Kondo resonances in a multilevel "semiopen" quantum dot. AB - Linear conductance across a large quantum dot via a single level epsilon(0) with large hybridization to the contacts is strongly sensitive to quasibound states localized in the dot and weakly coupled to epsilon(0). The conductance oscillates with the gate voltage due to interference of the Fano type. At low temperature and Coulomb blockade, Kondo correlations damp the oscillations on an extended range of gate voltage values, by freezing the occupancy of the epsilon(0) level itself. As a consequence, the antiresonances of Fano origin are washed out. The results are in good correspondence with experimental data for a large quantum dot in the semiopen regime. PMID- 15525195 TI - Exponential sensitivity to dephasing of electrical conduction through a quantum dot. AB - According to random-matrix theory, interference effects in the conductance of a ballistic chaotic quantum dot should vanish proportional to (tau(phi)/tau(D))(p) when the dephasing time tau(phi) becomes small compared to the mean dwell time tau(D). Aleiner and Larkin have predicted that the power law crosses over to an exponential suppression proportional to exp((-tau(E)/tau(phi)) when tau(phi) drops below the Ehrenfest time tau(E). We report the first observation of this crossover in a computer simulation of universal conductance fluctuations. Their theory also predicts an exponential suppression proportional to exp(( tau(E)/tau(D)) in the absence of dephasing--which is not observed. We show that the effective random-matrix theory proposed previously for quantum dots without dephasing explains both observations. PMID- 15525196 TI - New fluxon resonant mechanism in annular Josephson tunnel structures. AB - A novel dynamical state has been observed in the dynamics of a perturbed sine Gordon system. This resonant state has been experimentally observed as a singularity in the dc current-voltage characteristic of an annular Josephson tunnel junction, excited in the presence of a magnetic field. In this respect it can be assimilated to self-resonances known as Fiske steps. Differently from these, however, we demonstrate, on the basis of numerical simulations, that its detailed dynamics involves rotating fluxon pairs, a mechanism associated, so far, to self-resonances known as zero-field steps. This occurs because the size of nonlinear excitations is comparable with that of the system. PMID- 15525197 TI - Pair density wave in the pseudogap state of high temperature superconductors. AB - Recent scanning tunneling microscopy experiments of Bi(2)Sr(2)CaCu(2)O(8+delta) have shown evidence of real-space organization of electronic states at low energies in the pseudogap state [Science 303, 1995 (2004)]]. We argue based on symmetry considerations as well as model calculations that the experimentally observed modulations are due to a density wave of d-wave Cooper pairs without global phase coherence. We show that scanning tunneling microscopy measurements can distinguish a pair density wave from more typical electronic modulations such as those due to charge density wave ordering or scattering from an on site periodic potential. PMID- 15525198 TI - Coherent oscillations in a superconducting multilevel quantum system. AB - We have observed coherent oscillations in a multilevel quantum system, formed by a current-biased dc SQUID. These oscillations have been induced by applying resonant microwave pulses of flux. Quantum measurement is performed by a nanosecond flux pulse that projects the final state onto one of two different voltage states of the dc SQUID, which can be read out. The number of quantum states involved in the coherent oscillations increases with increasing microwave power. The dependence of the oscillation frequency on microwave power deviates strongly from the linear regime expected for a two-level system and can be very well explained by a theoretical model taking into account the anharmonicity of the multilevel system. PMID- 15525199 TI - Robust dx2-y2 pairing symmetry in hole-doped cuprate superconductors. AB - Although initially quite controversial, it is now widely accepted that the Cooper pairs in optimally doped cuprate superconductors have predominantly dx2-y2 wave function symmetry, and the controversy has now shifted to whether the pairing symmetry changes away from optimal doping. Here we present phase-sensitive tricrystal experiments on three cuprate systems: Y(0.7)Ca(0.3)Ba(2)Cu(3)O(7 delta) (Ca-doped Y-123), La2-xSrxCuO4 (La-214), and Bi(2)Sr(2)CaCu(2)O(8+delta) (Bi-2212), with doping levels covering the underdoped, optimal, and overdoped regions. Our work implies that predominantly d x2-y2 pairing symmetry is robust over a large variation in doping. PMID- 15525200 TI - Flux-line lattice distortion in PrOs4Sb12. AB - We report that the flux-line lattice in the cubic superconductor Pr(Os4Sb12 is strongly distorted from an ideal hexagonal lattice at very low temperatures in a small applied field. We attribute this to the presence of gap nodes in the superconducting state on at least some Fermi-surface sheets. PMID- 15525201 TI - muSR study of the quantum dynamics in the frustrated S=3/2 kagome bilayers. AB - We present muSR experiments in the S=3/2 kagome bilayer compound Ba(2)Sn(2)ZnGa(10-7p)Cr(7p)O22 [BSZCGO(p)] and compare it to the isostructural SrCr(9p)Ga(12-9p)O19 [SCGO(p)], including for the latter new results for p > or =0.89. Quantum-dynamical low energy magnetic excitations are evidenced in this novel compound. We study the evolution of the muon relaxation rate with p, T, and field. A phenomenological model for the muon relaxation based on sporadic dynamics due to spin excitations in a singlet sea proposed by Uemura et al. is extended to all fields and T range. Its connection to the RVB picture is discussed, and we argue that such coherent states might mediate the interactions between "impurities" which induce the spin glass freezing. PMID- 15525202 TI - Irradiation-induced magnetism in graphite: a density functional study. AB - Recent experiments indicate that proton irradiation triggers ferromagnetism in originally nonmagnetic graphite samples while He ion bombardment has a much smaller effect. To understand the origin of irradiation-induced magnetism, we have performed spin-polarized density functional theory calculations of the magnetic properties of the defects which are most likely to appear under irradiation vacancies and vacancy-hydrogen complexes. Both defects are magnetic, but as for the latter we find that H adsorption on one of the vacancy dangling bonds gives rise to a magnetic moment double that of the naked vacancy. We show that for small irradiation doses vacancy-hydrogen complexes result in a macroscopic magnetic signal which agrees well with the experimental values. PMID- 15525203 TI - Metamagnetic transition in Na 0.85 CoO2 single crystals. AB - We report the magnetization, specific heat, and transport measurements of a high quality Na(0.85)CoO2 single crystal in applied magnetic fields up to 14 T. At high temperatures, the system is in a paramagnetic phase. It undergoes a magnetic phase transition below approximately 20 K. For the field H||c, the measurement data of magnetization, specific heat, and magnetoresistance reveal a metamagnetic transition from an antiferromagnetic state to a quasiferromagnetic state at about 8 T at low temperatures. However, no transition is observed in the magnetization measurements up to 14 T for H perpendicular c. The low temperature magnetic phase diagram of Na(0.85)CoO2 is determined. PMID- 15525204 TI - Pressure and field induced magnetic order in the spin liquid Tb2Ti2O7 as studied by single crystal neutron diffraction. AB - We have studied the spin liquid Tb2Ti2O7 by single crystal neutron diffraction under high pressure up to 2.8 GPa, together with uniaxial stress, down to 0.1 K, in zero and high magnetic fields up to 7 T. In zero magnetic field, a long-range ordered antiferromagnetic structure is induced by pressure. The Neel temperature and ordered magnetic moment can be tuned by the anisotropic pressure component. Under magnetic field, the antiferromagnetic structure transforms into a canted ferromagnetic one at 0.6 T. Spin canting persists even at 7 T. The magnetic phase diagram under pressure shows a strong increase of the Neel temperature with the field. PMID- 15525205 TI - Low-energy singlets in the Heisenberg antiferromagnet on the kagome lattice. AB - The spin-1/2 Heisenberg antiferromagnet on the kagome lattice, is mapped by contractor renormalization to a spin-pseudospin Hamiltonian on the triangular superlattice. Variationally, we find a ground state with columnar dimer order. Dimer orientation fluctuations are described by an effective O(2) model at energies above an exponentially suppressed clock mass scale. Our results explain the large density of low-energy singlets observed numerically, and the nonmagnetic T2 specific heat observed experimentally. PMID- 15525206 TI - First-principles study of the electro-optic effect in ferroelectric oxides. AB - We first present a method to compute the electro-optic tensor from first principles, explicitly taking into account the electronic, ionic and piezoelectric contributions. We then study the nonlinear optic behavior of three paradigmatic ferroelectric oxides. Our calculations reveal the dominant contribution of the soft mode to the electro-optic coefficients in LiNbO3 and BaTiO3 and its minor role in PbTiO3. We identify the coupling between the electric field and the polar atomic displacements along the B-O chains as the origin of the large electro-optic response in perovskite ABO3 compounds. PMID- 15525207 TI - Structure of excited-state transitions of individual semiconductor nanocrystals probed by photoluminescence excitation spectroscopy. AB - We perform for the first time photoluminescence excitation (PLE) studies of individual nanocrystals (NCs) that reveal the structure of excited-state transitions not obscured by ensemble averaging. Single-NC PLE spectra strongly deviate from a traditional idealized picture of sharp, quasiatomic resonances. We detect only a few relatively narrow transitions (3-4 meV) at the band edge, while at higher spectral energies, we observe a broad structureless feature separated from the band-edge peaks by a >50 meV "minigap." These observations can be rationalized by analyzing hole intraband relaxation behavior. PMID- 15525208 TI - Two-photon lasers based on intersubband transitions in semiconductor quantum wells. AB - We propose to make a two-photon laser based on intersubband (sublevel) transitions in semiconductor nanostructures. The advantages and feasibility of such a two-photon laser are analyzed in detail using the density matrix approach. Both one-photon and two-photon gains in a three subband quantum well structure are studied on the same footing to show how the two-photon gain can be maximized, while the competing one-photon gain is minimized. The results show that a sufficient two-photon gain can be achieved to overcome one-photon competition and the loss of a conventional semiconductor cavity, making intersubband transitions one of the very few feasible approaches to two-photon lasing. PMID- 15525209 TI - Laser trapping of small colloidal particles in a nematic liquid crystal: clouds and ghosts. AB - We show that, contrary to intuition, small (< or =1 microm) transparent particles can be trapped and manipulated in a nematic liquid crystal using an intense laser beam, although their index of refraction is lower than both refractive indices of the surrounding birefringent fluid. Two mechanisms are identified that are responsible for this anomalous trapping: (i) surface-induced distortion of the birefringent media around the particle, creating a high-index "cloud" around the colloid, and (ii) laser-induced distortion or (partial) melting of a nematic, creating a ghost colloid. PMID- 15525210 TI - Theory of optical spectra involving charge transfer states: dynamic localization predicts a temperature dependent optical band shift. AB - The influence of charge transfer states on the optical line shape of chromophore complexes is investigated in a minimal model that includes a coupling between an excited state and an optically dark charge transfer state. In the calculations of the absorption spectrum, an intensity borrowing by the charge transfer state, strong vibrational sidebands, and a temperature dependent shift of optical transitions are obtained. The theory is applied to the bacterial photosynthetic reaction center to explain a 30 nm blueshift of the low-energy absorption band with increasing temperature. PMID- 15525211 TI - Scaling of F-actin network rheology to probe single filament elasticity and dynamics. AB - The linear and nonlinear viscoelastic response of networks of cross-linked and bundled cytoskeletal filaments demonstrates remarkable scaling with both frequency and applied prestress, which helps elucidate the origins of the viscoelasticity. The frequency dependence of the shear modulus reflects the underlying single-filament relaxation dynamics for 0.1-10 rad/sec. Moreover, the nonlinear strain stiffening of such networks exhibits a universal form as a function of prestress; this is quantitatively explained by the full force extension relation of single semiflexible filaments. PMID- 15525212 TI - Stability of membrane bound reactions. AB - We present a novel approach to the dynamics of reactions of diffusing chemical species with species fixed in space, e.g., by binding to a membrane. The nondiffusing reaction partners are clustered in areas with a diameter smaller than the diffusion length of the diffusing partner. The activated fraction of the fixed species determines the size of an active subregion of the cluster. Linear stability analysis reveals that diffusion is one of the major determinants of the stability of the dynamics. We illustrate the model concept with Ca2+ dynamics in living cells, which has release channels as fixed reaction partners. Our results suggest that spatial and temporal structures in intracellular Ca2+ dynamics are caused by fluctuations due to the small number of channels per cluster. PMID- 15525213 TI - Multiscale modeling of the surfactant mediated synthesis and supramolecular assembly of cobalt nanodots. AB - Multiscale simulations are used to bridge the surfactant templated assembly of individual approximately 1-10 nm cobalt dots, to their ordering into supramolecular arrays. Potential energy surfaces derived from ab initio calculations are input to lattice Monte Carlo simulations at atomic scales. By this process we quantitatively reproduce the experimental cobalt nanoparticle sizes. Crucially, we find that there is an effective short range attraction between pairs of nanodots. Mesoscale simulations show that these attractive interdot potentials are so short ranged that the dots can assemble only into orientally ordered hexatic phases as in the experiments. PMID- 15525214 TI - Excitable CO oxidation on Pt(110) under nonuniform coupling. AB - A new feedback scheme for guided spatiotemporal pattern formation in reaction diffusion systems is introduced. In contrast to previously established control methods, we present a coupling protocol that is sensitive to the presence of coherent structures in the medium. Applying this feedback to the catalytic CO oxidation on Pt(110) in both experiments and numerical simulations, we show that temporal evolution and spatial extension of self-organizing objects can be efficiently controlled. PMID- 15525215 TI - Fitness-dependent topological properties of the world trade web. AB - Among the proposed network models, the hidden variable (or good get richer) one is particularly interesting, even if an explicit empirical test of its hypotheses has not yet been performed on a real network. Here we provide the first empirical test of this mechanism on the world trade web, the network defined by the trade relationships between world countries. We find that the power-law distributed gross domestic product can be successfully identified with the hidden variable (or fitness) determining the topology of the world trade web: all previously studied properties up to third-order correlation structure (degree distribution, degree correlations, and hierarchy) are found to be in excellent agreement with the predictions of the model. The choice of the connection probability is such that all realizations of the network with the same degree sequence are equiprobable. PMID- 15525216 TI - Nanostructures in biodefense and molecular diagnostics. PMID- 15525218 TI - Advances in genome biology and technology. PMID- 15525219 TI - QA/QC: challenges and pitfalls facing the microarray community and regulatory agencies. AB - The scientific community has been enthusiastic about DNA microarray technology for pharmacogenomic and toxicogenomic studies in the hope of advancing personalized medicine and drug development. The US Food and Drug Administration has been proactive in promoting the use of pharmacogenomic data in drug development and has issued a draft guidance for the pharmaceutical industry on data submissions. However, many challenges and pitfalls are facing the microarray community and regulatory agencies before microarray data can be reliably applied to support regulatory decision making. Four types of factors (i.e., technical, instrumental, computational and interpretative) affect the outcome of a microarray study, and a major concern about microarray studies has been the lack of reproducibility and accuracy. Intralaboratory data consistency is the foundation of reliable knowledge extraction and meaningful crosslaboratory or crossplatform comparisons; unfortunately, it has not been seriously evaluated and demonstrated in every study. Profound problems in data quality have been observed from analyzing published data sets, and many laboratories have been struggling with technical troubleshooting rather than generating reliable data of scientific significance. The microarray community and regulatory agencies must work together to establish a set of consensus quality assurance and quality control criteria for assessing and ensuring data quality, to identify critical factors affecting data quality, and to optimize and standardize microarray procedures so that biologic interpretation and decision-making are not based on unreliable data. These fundamental issues must be adequately addressed before microarray technology can be transformed from a research tool to clinical practices. PMID- 15525220 TI - Emergent FDA biodefense issues for microarray technology: process analytical technology. AB - A successful biodefense strategy relies upon any combination of four approaches. A nation can protect its troops and citizenry first by advanced mass vaccination, second, by responsive ring vaccination, and third, by post-exposure therapeutic treatment (including vaccine therapies). Finally, protection can be achieved by rapid detection followed by exposure limitation (suites and air filters) or immediate treatment (e.g., antibiotics, rapid vaccines and iodine pills). All of these strategies rely upon or are enhanced by microarray technologies. Microarrays can be used to screen, engineer and test vaccines. They are also used to construct early detection tools. While effective biodefense utilizes a variety of tactical tools, microarray technology is a valuable arrow in that quiver. PMID- 15525221 TI - NMP22 BladderChek Test: point-of-care technology with life- and money-saving potential. AB - A new, relatively obscure tumor marker assay, the NMP22 BladderChek Test (Matritech, Inc.), represents a paradigm shift in the diagnosis and management of urinary bladder cancer (transitional cell carcinoma). Specifically, BladderChek should be employed every time a cystoscopy is performed, with corresponding changes in the diagnostic protocol and the guidelines of the American Urological Association for the diagnosis and management of bladder cancer. Currently, cystoscopy is the reference standard and NMP22 BladderChek Test in combination with cystoscopy improves the performance of cystoscopy. At every stage of disease, BladderChek provides a higher sensitivity for the detection of bladder cancer than cytology, which now represents the adjunctive standard of care. Moreover, BladderChek is four-times more sensitive than cytology and is available at half the cost. Early detection of bladder cancer improves prognosis, quality of life and survival. BladderChek may be analogous to the prostate-specific antigen test and eventually expand beyond the urologic setting into the primary care setting for the testing of high-risk patients characterized by smoking history, occupational exposures or age. PMID- 15525222 TI - Computational analysis of gene-gene interactions using multifactor dimensionality reduction. AB - Understanding the relationship between DNA sequence variations and biologic traits is expected to improve the diagnosis, prevention and treatment of common human diseases. Success in characterizing genetic architecture will depend on our ability to address nonlinearities in the genotype-to-phenotype mapping relationship as a result of gene-gene interactions, or epistasis. This review addresses the challenges associated with the detection and characterization of epistasis. A novel strategy known as multifactor dimensionality reduction that was specifically designed for the identification of multilocus genetic effects is presented. Several case studies that demonstrate the detection of gene-gene interactions in common diseases such as atrial fibrillation, Type II diabetes and essential hypertension are also discussed. PMID- 15525223 TI - Biomarkers of cardiac disease. AB - The challenge of medical practice today is to identify individuals who are at risk of developing disease, determine the severity of the disease and distinguish the responders from the nonresponders to therapy (individualized medicine). Advances in molecular genetics and biology have shifted the paradigm for identification of markers from large-scale epidemiologic studies to studies on genomic- and proteomic-based techniques. Consequently, a large number of biologic markers, referred to as biomarkers, are being identified and validated to serve for risk stratification, prognostication and individualization of therapy. Identification of biomarkers for cardiovascular diseases could also provide insight into the pathogenesis of the phenotype, which is fundamental for the development of specific therapies. The list of biomarkers for cardiovascular disease is expanding rapidly. Nonetheless, the field is in the early stages of evolution and large-scale clinical studies are required to validate the utility of newly identified biomarkers in diagnosis, risk stratification and treatment of cardiovascular diseases. Selected biomarkers for coronary atherosclerosis, acute coronary syndromes and heart failure are discussed in this review. PMID- 15525224 TI - Mass spectrometry for malaria diagnosis. AB - A physical method currently being developed for malaria parasite detection and diagnosis in blood is reviewed in this article. The method - direct laser desorption mass spectrometry - is based on the detection of heme (iron protoporphyrin) as a unique qualitative and quantitative molecular biomarker for malaria. In infected erythrocytes, the parasite sequesters heme in a molecular crystal (hemozoin) - a volume of highly concentrated and purified biomarker molecules. Laser desorption mass spectrometry detects only heme from hemozoin in parasite-infected blood, and not heme that is bound to hemoglobin or other proteins in uninfected blood samples. The method requires only a drop of blood with minimal sample preparation. Laser desorption mass spectrometry may become a rapid and high-throughput tool for specific and sensitive pan-malaria detection at levels below 10 parasites/mul of blood. PMID- 15525225 TI - Laser capture microdissection, microarrays and the precise definition of a cancer cell. AB - Most expression profiling studies of solid tumors have used biopsy samples containing large numbers of contaminating stromal and other cell types, thereby complicating any precise delineation of gene expression in nontumor versus tumor cell types. Combining laser capture microdissection, RNA amplification protocols, microarray technologies and our knowledge of the human genome sequence, it is possible to isolate pure populations of cells or even a single cell and interrogate the expression of thousands of sequences for the purpose of more precisely defining the biology of the tumor cell. Although many of the studies that currently allow for characterization of small sample preparations and single cells were performed utilizing noncancer cell types, and in some cases isolation protocols other than laser capture microdissection, a list of protocols are described that could be used for the expression analysis of individual tumor cells. Application of these experimental approaches to cancer studies may permit a more accurate definition of the biology of the cancer cell, so that ultimately, more specific targeted therapies can be developed. PMID- 15525226 TI - Multiplex diagnostic platforms for detection of biothreat agents. AB - The availability of rapid, sensitive and cost-effective diagnostic methods is paramount to the success of a comprehensive national health security system in the USA. The national networks that were established to safeguard US infrastructures (e.g., public health, livestock, agriculture and water supply) have developed sufficient capability and capacity for monitoring. However, additional advanced methods will be required to maintain operational readiness. Currently available methods, although sensitive and specific, are generally costly and not amenable to high-throughput analyses. Critical to the success of biothreat surveillance is the ability to screen for and detect multiple agents rapidly in a single reaction and with minimal sample processing. This review will examine currently available diagnostic platforms (i.e., PCR-, immuno- and array based) and biosensors that can detect multiple biothreat analytes in a single reaction (i.e., multiplex assays). The maturity, benefits and limitations of each platform will be described and a prospective view, from current to future state of the art, will be proposed. PMID- 15525227 TI - Molecular recognition of biowarfare agents using micromechanical sensors. AB - Recent terrorist events have demonstrated that an urgent and widespread need exists for the development of novel sensors for threat detection, especially biowarfare agents. The advent of inexpensive, mass-produced microcantilever sensors promises to bring about a revolution in detection of terrorist threats. Extremely sensitive and highly selective sensors can be developed for using a microcantilever platform. Microcantilevers undergo bending when molecules are adsorbed on a single side. For biowarfare agent detection, specificity is achieved by immobilizing antibodies on one side of the cantilever. Antigen adsorption decreases surface energy and stress, resulting in cantilever deflection. PMID- 15525228 TI - A tale of a few hospitals. PMID- 15525230 TI - Falls are a sensitive and challenging area of health care. PMID- 15525229 TI - Let's ban drivel: commentary on A tale of a few hospitals. PMID- 15525231 TI - Public and private intervention rates in obstetric practice. PMID- 15525232 TI - Commentary: Refashioning child and family health services in response to family, social and political change. PMID- 15525233 TI - Tensions in maternal and child health policy in Victoria: looking back, looking forward. AB - Since the late 1980s, Maternal and Child Health Services (MCHS) in Victoria have undergone significant change. This paper provides an historically-informed analysis of the complex intersection of policy, administrative restructuring and stakeholder interests. It draws on and extends the authors' previous research into MCH Service policy directions and administration, including the impact of Compulsory Competitive Tendering (CCT) on MCH nurses in the 1990s. Historically there has been little explicit debate about either organisational arrangements, or the policy objectives of the MCHS. The dominant focus on health surveillance of infants never adequately reflected nurses' wider role in the community and was not consistent with a wider social model of health. Tensions between professional, consumer and administrative stakeholders became heightened by the implementation of the 1990s neoliberal political agenda. During this period, when restructuring linked funding to service delivery through tendering arrangements, a political and policy settlement further institutionalised surveillance as the basis of the MCHS. The restructured Service has remained constrained by the dominance of health surveillance as the primary program goal even after more varied contracting arrangements replaced CCT. Although recent initiatives indicate signs of hange, narrow surveillancebased guidelines for Victorian MCH Services are not consistent, we argue, with recent early years of life policy which calls for approaches derived from socio-ecological models of health. PMID- 15525234 TI - A preliminary case-mix classification system for Home and Community Care Clients in Western Australia. AB - political and policy settlement further institutionalised surveillance as the basis of the MCHS. The restructured Service has remained constrained by the dominance of health surveillance as the primary program goal even after more varied contracting arrangements replaced CCT. Although recent initiatives indicate signs of hange, narrow surveillancebased guidelines for Victorian MCH Services are not consistent, we argue, with recent early years of life policy which calls for approaches derived from socio-ecological models of health. PMID- 15525235 TI - Demographics and utilisation of health services by paediatric refugees from East Africa: implications for service planning and provision. AB - Little is known of difficulties in accessing health care for recently arrived paediatric refugees in Australia. We reviewed routinely collected data for all 199 East African children attending a hospital Immigrant Health Clinic for the first time over a 16 month period. Although 63% of parents reported medical consultations since arrival, 77% of this group reported outstanding, unaddressed health problems. Availability of interpreters and information on health services were the main factors hindering access to care. These data have informed future service planning at the Clinic. Ongoing data collection is key to maintaining a responsive, targeted service for a continually changing population. PMID- 15525236 TI - The lived experience of men and women with hepatitis C: implications for support needs and health information. AB - routinely collected data for all 199 East African children attending a hospital Immigrant Health Clinic for the first time over a 16 month period. Although 63% of parents reported medical consultations since arrival, 77% of this group reported outstanding, unaddressed health problems. Availability of interpreters and information on health services were the main factors hindering access to care. These data have informed future service planning at the Clinic. Ongoing data collection is key to maintaining a responsive, targeted service for a continually changing population. PMID- 15525237 TI - Hepatitis C-related discrimination in New South Wales. AB - Hepatitis C-related discrimination was examined. Intake interviews with 606 HepCare trial participants from New South Wales were analysed to determine the prevalence and correlates of hepatitis C related discrimination. The sample was a mean age of 37 years, 54% were males, 79% reported a history of drug injecting and 35% were current injectors. Forty percent of the sample reported experiencing hepatitis C-related discrimination. Multivariate analysis revealed that current injectors, 35-44 year olds, females, those who had recently consulted a general practitioner and those who had been referred to a specialist for their hepatitis C were more likely to report discrimination than other groups. More research is required to attain a better understanding of hepatitis C-related discrimination. PMID- 15525238 TI - Demographic patterns of emergency presentations to Northern Territory public hospitals. AB - This study investigates demographic patterns of emergency presentations to Northern Territory (NT) public hospitals over the past five years with respect to population changes, Aboriginality and age of patients. Retrospective analysis was undertaken on the 1996-2001 data extracted from the NT Module of Caresys and the Hospital Morbidity Data System. There was a 4.6% decrease in total presentations to the five public hospitals but a 9.4% growth in the population during the study period. Substantial differences in emergency presentation patterns were found between Aboriginal and non-Aboriginal patients. There were more Aboriginal presentations than non-Aboriginal presentations for all age groups except for 5 to 19 years and 70 to 74 years. Analysis based on the national triage scale showed the higher needs of older adults with the 60 or over age group accounting for the majority of presentations, and Aboriginal presentation rates exceeded the non-Aboriginal presentation rates in most triage categories. Re-attendance within seven days at the emergency departments occurred predominantly among Aboriginal patients regardless of age group. The analysis has highlighted several emerging demographic patterns. The issue of non-urgent visits by Aboriginal patients occupying a large portion of the emergency department utilisation also needs to be addressed. PMID- 15525239 TI - The influence of socio-economic and locational disadvantage on patterns of surgical care for lung cancer in Western Australia 1982-2001. AB - OBJECTIVE: Patterns of in-hospital surgical care for lung cancer in Western Australia were examined, including the effects of demographic, locational and socio-economic disadvantage and the possession of private health insurance, on the likelihood of receiving surgery. PATIENTS AND METHODS: The WA Record Linkage Project was used to extract hospital morbidity, cancer and death records of all people with lung cancer in Western Australia from 1982 to 2001. The likelihood of receiving lung cancer surgery was estimated, after adjustment for co-variates, using logistic regression. RESULTS: Overall, 16% of patients received surgery for their lung cancer, although this varied according to histology. Patients who received surgery were typically younger, female, non-indigenous and had less comorbidity. Patients from socio-economically disadvantaged groups tended to be less likely to receive surgery (OR 0.79; 95% CI 0.61-1.04) although this was not significant for each category of disadvantage. Those who had their first hospital admission, with a mention of lung cancer, in a rural hospital were less likely to receive surgery (OR 0.26; 95% CI 0.19-0.36) than those in metropolitan hospitals, although residential location generally had less effect (OR 0.36; 95% CI 0.14 0.92). Patients admitted as a private patient either to a private or public hospital for their first mention of lung cancer had increased likelihood of receiving surgery (OR 1.15; 95% CI 1.02-1.30); however first admission to a private hospital had no effect (OR 0.99: 95% CI 0.85-1.16). CONCLUSION: The utilisation of lung cancer surgery was low with several factors found to affect the rate. Patients from socio-economically or locationally disadvantaged backgrounds, indigenous patients or patients without private health insurance were less likely to receive lung cancer surgery than those from more advantaged groups. PMID- 15525240 TI - The impact of manual handling training on work place injuries: a 14 year audit. AB - Manual handling injuries represented both the highest costs and largest proportion of workers compensation claims for the South Australian Department of Human Services from 1995 to 1999 (Risk Management Services 2000). There is evidence to suggest the manutention method of manual handling training can reduce workplace injuries. A retrospective audit of injuries was conducted at Metropolitan Domiciliary Care Southern Region, a community health service agency, to determine the effects of introducing the manutention method of manual handling to the agency. The workers most likely to sustain a manual handling injury were paramedical aides. For this workgroup, the incidence and severity of injuries were significantly reduced for the period following the introduction of the manutention method of manual handling training. PMID- 15525241 TI - Australian women's perceptions of a specialist breast nurse model. AB - The aim of this study was to evaluate Australian women's perceptions of the specialist breast nurse (SBN) model developed by the National Breast Cancer Centre. Two hundred and forty women evaluated the care they received from a SBN. The SBN was perceived as a valuable link between women and the multi-disciplinary team, with continuity of care being rated as a major benefit. Over 80% of women reported the SBN to be effective in providing information and emotional support. There is a need to consider the establishment of SBN positions in treatment centres for breast cancer in Australia. PMID- 15525242 TI - Sustaining evidence-based practice for young people who self-harm: a 4-year follow-up. AB - OBJECTIVE: In 1998 and 1999, two NSW Area Health Services conducted the Youth At Risk of Deliberate Self Harm (YARDS) project. The YARDS project was designed to implement evidence-based service enhancements for the clinical management of young people with Deliberate Self Harm. This paper examines the extent to which service enhancements implemented during YARDS were maintained 4 years after the conclusion of the project and compares service quality with another NSW Area Health Service that did not participate in the YARDS project. METHOD: Staff from mental health services and emergency departments completed the Service Activity Scale, a measure developed for the YARDS project to assess the quality of health service response to individuals who present following a suicide attempt. RESULTS: RESULTS indicated that the service improvements made during the YARDS project were maintained 4years after the project ended. Furthermore, a significant difference was found between scores for services that participated in YARDS and services that did not participate in YARDS. CONCLUSIONS: These results suggest that projects such as YARDS which support evidence based service nhancements may be useful in improving the management of young people with deliberate self-harm, and that these improvements may be long-lasting. PMID- 15525243 TI - People who are 'socially disadvantaged' and the role of the aged care assessment team: a case example. PMID- 15525244 TI - A new look for Australian Health Review. PMID- 15525245 TI - The weakest link? PMID- 15525246 TI - Patient safety, ethics and whistleblowing: a nursing response to the events at the Campbelltown and Camden Hospitals. PMID- 15525247 TI - What's wrong with our hospitals? PMID- 15525248 TI - Peer review of adverse events - a perspective on Macarthur. AB - Recent investigations into the Macarthur Health Service have resulted in multiple reviews of a small number of cases. This article was prompted by a casual observation that these reviews have resulted in differing conclusions about what occurred in each case and what might have been done in response. The reliability of peer review is examined, together with the literature on the scale of adverse events and the issue of problem identification. Potential sources of bias and error during peer review are considered. Drawing on the lessons from the literature and the experience of Macarthur, suggestions are made to improve the identification and review of adverse events. PMID- 15525249 TI - Private health insurance and the Pharmaceutical Benefits Scheme: how effective has recent government policy been? AB - The cost to government of the Pharmaceutical Benefits Scheme (PBS) is rising at over 10 percent per annum. The government subsidy to Private Health Insurance (PHI) is about $2.4 billion and rising. Despite this, the queues facing public patients - which were the primary justification for the assistance to PHI - do not appear to be shortening. Against this backdrop, we seek to evaluate recent policies. It is shown that the reason commonly given for the support of PHI - the need to preserve the market share of private hospitals and relieve pressure upon public hospitals - is based upon a factually incorrect analysis of the hospital sector in the last decade. It is similarly true that the 'problem' of rising pharmaceutical expenditures has been exaggerated. The common element in both sets of policies is that they result in cost shifting from the public to the private purse and have little to do with the quality or quantity of health services. PMID- 15525250 TI - Why protect confidentiality in health records? A review of research evidence. AB - We present the main arguments for protecting the confidentiality of health services, along with those for limiting confidentiality. These arguments are then substantiated by reference to research evidence. There is evidence that access to health care is restricted if confidentiality is not promised to some groups of patients. Fear of disclosure does diminish patients' candour, and this can compromise the quality of care. While patients are concerned about confidentiality and some are harmed by 'leaks' from health services, most people in Australia still trust health providers to keep their secrets, and patients rarely become aware of a breach of confidence. It has been claimed that strict protection of confidentiality may obstruct the pursuit of medical research and the use of electronic medical records. There is, as yet, no evidence that gaining full benefit from the use of electronic medical records entails reduced protection for confidentiality. The losses to epidemiological research if patient consent were always required are hotly debated. Confidentiality should be protected because it protects patients from harm, supports access to health care and produces better health outcomes. PMID- 15525251 TI - How empowerment and social support affect Australian nurses' work stressors. AB - We surveyed 157 nurses at a private hospital in Melbourne to examine the role of social support and empowerment in reducing work stress among Australian nurses. Our findings reveal that social support, derived from either the supervisor or work colleagues, is negatively associated with the main work stressors, such as role conflict, role ambiguity, work overload and resource inadequacy. Empowerment, too, is negatively associated with the main work stressors, apart from work overload. We discuss contributions of this study and implications for research and practice in health sector management. PMID- 15525252 TI - A survey of workplace bullying in a health sector organisation. AB - The purpose of this study was to explore the frequency, nature and extent of workplace bullying in a health care organisation in New South Wales. The survey methodology achieved a 79% response rate (n = 311). Overall, 155 respondents (50%) reported that they had experienced one or more forms of bullying behaviour in the past 12 months. The largest reported source of workplace bullying was peers or fellow workers (49%), followed by clients (42%) and managers or supervisors (38%). Only 36% of respondents who had been bullied had formally reported the episode. The level of bullying reported was unexpectedly high and will require development of strategies to address the issue. PMID- 15525253 TI - Gender differences in senior management decision making in the New England Area Health Service. AB - Data from a questionnaire administered to senior managers in the New England Area Health Service (NEAHS) was used to examine gender differences in decision-making processes. The study found that female managers were more likely to report that they included staff in decision-making processes. The small size of the population restricted the statistical analysis; more meaningful findings may result if the study were to be repeated using a larger population of senior managers. PMID- 15525254 TI - Reducing utilisation of hospital services by case management: a randomised controlled trial. AB - A 12-month randomised controlled trial was conducted in Hong Kong to evaluate the effectiveness of case management provided to a group of home-dwelling, frail elderly patients (control group: n = 47; intervention group: n = 45) in terms of utilisation of hospital services by these patients. Significant reductions (significance at P 1.35, the number of known genes was 33 (38%), and the number of expressed sequence tag (EST) sequences was 55 (62%). Among the 130 genes for which the contra : ipsi was > 1.35, the number of known genes was 55 (42%), and the number of EST sequences was 75 (58%). Changes in some of the genes were consistent with previous studies; however, we found several new genes which could be functionally related to the molecular basis of the electrophysiological asymmetry between the VNCs following UVD. Ipsi > contra genes included the GABA(A) receptor rho subunit, regulatory proteins of G protein signaling, calcium signaling related molecules such as the voltage-dependent calcium channel alpha2/delta subunit 1, calcineurin subunit Abeta and Ca(2+) pump. Contra > ipsi genes included the neuronal high affinity glutamate transporter, 5-hydroxytryptamine receptor 1D, mitogen-activated protein kinase 12 and ubiquitin carboxy-terminal hydrolase L1. PMID- 15525352 TI - Two different Ca2+-dependent inhibitory mechanisms of spontaneous firing by glutamate in dopamine neurons. AB - The excitatory neurotransmitter, glutamate, generates a characteristic burst pause type of firing in midbrain dopamine neurons in association with the reward behavior, but the cellular mechanism by which glutamate generates these bursts is unknown. Here, we show that the bursts in spontaneously firing dopamine neurons can be generated by the combinative actions of the brief stimulatory and the subsequent Ca(2+)-dependent inhibitory signals in response to glutamate stimulation. The two Ca(2+)-dependent firing-extinction signals are activated by different glutamate receptors. Although the activation of metabotropic glutamate receptors rapidly stopped the enhanced firing through the Ca(2+) release from intracellular stores, the activation of NMDA and AMPA/kainate receptors abolished the firing immediately after termination of the stimulation due to the Ca(2+) accumulation in the cell. These two Ca(2+)-dependent inhibitory mechanisms appear to participate in the generation of characteristic bursts in dopamine neurons by controlling the maximum firing number of single bursts and the duration of post firing pauses. PMID- 15525353 TI - A proteasomal stress response: pre-treatment with proteasome inhibitors increases proteasome activity and reduces neuronal vulnerability to oxidative injury. AB - We report here that exposure to low concentrations of proteasome inhibitors (e.g. 10-100 nm MG-132, 0.1-3 nm epoxomicin or 10-30 nm clasto-lactacystin beta lactone) resulted in an enhancement, rather than an inhibition, of proteasome activity in cultured neocortical neurons. Size-fractionation chromatography confirmed that the enhanced peptide cleavage activity was associated with proteasome-sized complexes. This sub toxic exposure reduced neuronal death caused by subsequent exposure to oxidative stress (100-200 microm H(2)O(2) for 30 min, 24-h exposure to 100 microm paraquat or 7.5 microm menadione), but did not alter vulnerability to excitotoxicity (5-min exposure to 30-100 microm NMDA or 24 exposure to 12 microm NMDA). Sub toxic proteasome inhibitor exposure caused an increase in levels of proteasome core subunit proteins and mRNAs, but not in levels of potentially cytoprotective heat shock proteins (hsp70, hsp90 and hsp40). The neuroprotective effects of proteasome inhibitor pre-treatment were blocked by coapplication of proteasome inhibitors during the oxidative insult. These findings support a model in which sublethal proteasome inhibition induces neurons to increase proteasome activity and promotes resistance to oxidative injury and suggests that enhancement of proteasome activity is a potential therapeutic target for diseases in which oxidative stress has been implicated. PMID- 15525354 TI - Cloning of a novel neuronally expressed orphan G-protein-coupled receptor which is up-regulated by erythropoietin, interacts with microtubule-associated protein 1b and colocalizes with the 5-hydroxytryptamine 2a receptor. AB - G-protein-coupled receptors (GPCRs) are the largest group of cell surface molecules involved in signal transduction and are receptors for a wide variety of stimuli ranging from light, calcium and odourants to biogenic amines and peptides. It is assumed that systematic genomic data-mining has identified the overwhelming majority of all remaining GPCRs in the genome. Here we report the cloning of a novel orphan GPCR which was identified in a search for erythropoietin-induced genes in the brain as a strongly up-regulated gene. This unknown gene coded for a protein which had a seven-transmembrane topology and key features typical of GPCRs of the A family but a low overall identity to all known GPCRs. The protein, coded ee3, has an unusually high evolutionary conservation and is expressed in neurons in diverse areas of the CNS with relation to integrative functions or motor tasks. A yeast two-hybrid screen for interacting proteins revealed binding to the microtubule-associated protein (MAP) 1b. Coupling to MAP1a has been described for another cognate GPCR, the 5 hydroxytryptamine (5HT) 2a receptor. Surprisingly, we found complete colocalization of ee3 and the 5HT2a receptor. The interaction with MAP1b proved to be critical for the stability or folding of ee3 as in mice lacking MAP1b the ee3 protein was undetectable by immunohistochemistry, although messenger RNA levels remained unchanged. We propose that ee3 is a highly interesting new orphan GPCR with potential connections to erythropoietin and 5HT2a receptor signalling. PMID- 15525355 TI - Recombinant DNA vaccine encoding multiple domains related to inhibition of neurite outgrowth: a potential strategy for axonal regeneration. AB - Myelin-derived proteins, such as tenascin-R (TN-R), myelin associate glycoprotein (MAG), and Nogo-A, inhibit the CNS regeneration. By targeting specifically the inhibitory epitopes, we have investigated whether vaccination with a recombinant DNA molecule encoding multiple domains of myelin inhibitors may be useful in CNS repair. We show here that the recombinant DNA vaccine is able to activate the immune system but does not induce experimental autoimmune encephalomyelitis (EAE) in Lewis rats. Importantly, it promotes axonal regeneration in a spinal cord injury model. Thus, the application of DNA vaccine, encoding multiple specific domains of major inhibitory proteins and/or their receptors, provides another promising approach to overcome the inhibitory barriers during CNS regeneration. PMID- 15525357 TI - Melioidosis in animals: a review on epizootiology, diagnosis and clinical presentation. AB - Melioidosis, an infectious disease caused by Burkholderia pseudomallei is an emerging disease with high impact on animals and man. In different animal species, the clinical course varies and delayed diagnosis poses risks for the dissemination of the agent in non-endemic areas. Not only migration and transport of animals around the world but also tourism increases the risk that melioidosis can leave its endemic boundaries and establish itself elsewhere. Detection of the agent is a major challenge, as the agent has to be handled in laboratories of biosafety level 3 and test kits are not yet commercially available. Veterinarians and doctors should be aware of melioidosis not only as an agent of public interest but also in terms of a bioterrorist attack. The aim of this review is to describe the agent, its aetiology, the manifestation in a variety of animal species as well as to describe diagnostic procedures, typing techniques and countermeasures. PMID- 15525358 TI - Risk factors for mortality in grow-finishing pigs in Belgium. AB - The present study investigated risk factors for mortality in grow-finishing pigs of 137 pig herds belonging to one integration company during a period of 2.5 years. Mortality data, expressed as the number of dead pigs divided by the number of pigs placed in the fattening unit were investigated retrospectively. The following potential risk factors were evaluated: type of pig herd, season and year of placement in the fattening unit, pig density in the municipality, management practices (density of the pigs in the barn, origin of the pigs), housing conditions and feeding practices. The overall average mortality percentage was 4.70%. Three variables in a multivariable regression model were significantly associated with mortality: season of placement in the fattening unit, origin of the piglets and duration of the fattening period. Pigs placed in October, November and December, were at higher risk than pigs placed in other months. Herds that purchased pigs from a merchant, used pigs from other herds with an excess of piglets for filling their fattening units or herds that purchased pigs from more than five origin herds also suffered higher mortality. The mortality also increased in case of longer duration of the fattening period. This study documented for the first time that in addition to seasonal effects and a longer duration of the fattening period, purchasing feeder pigs from one or a limited number of herds is of crucial importance to achieve low mortality in the grow-finishing pigs. PMID- 15525359 TI - Rabies in red foxes (Vulpes vulpes) experimentally infected with European bat lyssavirus type 1. AB - The susceptibility of red foxes (Vulpes vulpes) to European bat lyssavirus type 1 (EBLV-1) infection was examined. Eight foxes were inoculated intramuscularly (i.m.) with 10(4.9) foci-forming units (FFU) (n = 4) and 10(5.1) FFU (n = 4) and observed for up to 90 days. All foxes showed manifestations of a neurologic disorder (e.g. seizures, myoclonus, agitation), starting as early as 5 days post infection (p.i.). Subsequently, all animals showed improvement followed by one or more relapses. One fox was killed 3 days after it recovered, 26 days post infection. Two other foxes were also killed 38 and 54 days post-infection after severe neurologic signs returned. All foxes developed a humoral immune response against EBLV-1 as determined in serum and brain tissues. However, no rabies virus antigen was detected in the brain, other tissues and secretions examined (e.g. salivary gland, saliva, tonsils, lungs) by using different standard diagnostic techniques [fluorescent antibody test, reverse transcription polymerase chain reaction (RT-PCR), rabies tissue culture inoculation test], with the exception of one fox in which EBLV-1 RNA was detected by RT-PCR in only the spinal cord. Brain tissues showed moderate to severe multifocal, mononuclear encephalomyelitis in the three foxes that were killed during the observation period, although no EBLV 1 virus was detectable in these tissues. PMID- 15525360 TI - Epidemiological study of non-contagious intramammary infections in nine commercial dairy herds following a Staphylococcus aureus control programme. AB - Changes in prevalence in intramammary infection, by pathogen type, in herds applying a stringent contagious mastitis control programme was studied. Enrollment of 1651 lactating cows and collection of milk samples was made in this ancillary study to a cohort study of the dynamics of mastitis prevalence after adoption of a strict contagious mastitis control programme that targeted the elimination of mastitis caused by Staphylococcus aureus. Nine commercial dairies in Italy were used. Aseptic collection of milk samples from all lactating cows was performed at the time of enrollment, from all cows within 7-14 days of entering the lactating herd after the date of enrollment, and from all lactating cows at 2, 4, 7, 10 and 14 months after the date of enrollment. Prevalence of intramammary infection by pathogen type was determined from culture of milk samples. Application of the strict contagious mastitis programme did not lead to an increased risk of non-contagious mastitis. The risk of coliform, environmental streptococcal and coagulase-negative staphylococcal intramammary infections decreased after adoption of the programme. The data reported herein indicate that the overall risk for any intramammary infections decreases with adoption of a strict contagious mastitis programme, and that such a programme therefore does not necessarily lead to an increase in environmental mastitis. PMID- 15525361 TI - Prevalence and antimicrobial resistance of Salmonella spp. in non-diarrhoeic dogs in Trinidad. AB - The estimated prevalence and antimicrobial resistances of Salmonella spp. in non diarrhoeic dogs across Trinidad was determined. The serotypes of Salmonella spp. isolated were also identified. Of a total of 1391 dogs sampled, 50 (3.6%) were positive for Salmonella spp. with 28 different serotypes, the predominant serotypes were Javiana (12), Newport (6), Arechavaleta (5) and Heidelberg (5). Fifty-seven (85.1%) of 67 isolates exhibited resistance to one or more antimicrobial agents. Of eight antimicrobial agents tested, resistance was exhibited to streptomycin (80.6%), cephalothin (37.3%), neomycin (38.8%) and gentamicin (9.0%). All isolates were sensitive to ampicillin, norfloxacin, choramphenicol and sulphamethoxazole/trimethoprim. It was concluded that the isolation of the Salmonella spp. from non-diarrhoeic dogs could pose health hazard to their owners as most serotypes are known to be virulent. Furthermore, the prevalence of resistance to antimicrobial agents amongst the Salmonella isolates from these animals indicates susceptibility testing may influence chemotherapeutic choices when treating these isolates. PMID- 15525362 TI - Distribution of virulence genes in Escherichia coli strains isolated from diarrhoeic piglets in the Slovak Republic. AB - Ninety-two Escherichia coli isolates from 14 to 28-day-old piglets that died because of diarrhoea were examined for genes for fimbriae (F4, F5, F6, F18 and F41), enterotoxins (STa, STb and LT), verotoxin (VT2e or Stx2e) and enteroaggregative heat-stable enterotoxin 1 (EAST1) by polymerase chain reaction. Twenty-two strains (24%) carried a gene for F4, whereas genes for F18, F6 and F5 + F41 were detected in 10.8, 3.3 and 1.1% of strains respectively. Genes for STb, LT, STa and Stx2e were detected in 40.2, 26.1, 14.1 and 1.1% of strains respectively. The astA gene was detected in 49 (53.3%) isolates, 35 of which also carried genes for enterotoxins and/or fimbriae. The major genotypes reached at (in decreasing order of prevalence) were F4/STb/LT/EAST1, F18/STa/STb/EAST1, STb/EAST1, F6/STa/STb/EAST1 and F18/STb/EAST1. PMID- 15525363 TI - Neospora-associated abortion episode over a 1-year period in a dairy herd in north-east Spain. AB - This report describes a retrospective study based on serological screening, performed on a Neospora-associated abortion episode over a 12-month period in a dairy herd in north-east Spain. During the 1-year study period, the overall abortion rate for the herd was 23.2% (38 abortions of 164 diagnosed pregnancies). The data analysed were those derived from blood samples collected from the whole herd (n = 237) and from diagnosed pregnancies. Antibodies to Neospora caninum were found in 35.4% of the cattle with 44% of seropositive pregnant animals aborting over 1-year period. Based on the odds ratio, the risk of abortion was 12.2 times higher (P < 0.0001) in the Neospora-seropositive animals than in seronegative animals and significantly higher during the second term of gestation (P < 0.01) than during the first and third terms. Abortions were not found to be associated with parity status or season of pregnancy, and the common risk factors associated with pregnancy loss in the geographical area of the study. Age-related differences in N. caninum seroprevalence were not statistically significant, indicating vertical transmission as the main route of infection. Indeed, a high percentage of congenitally infected offspring was observed (90.6%) and the farm had been free of dogs for the last 7 years. Our results suggest that, when a dairy herd shows an increased incidence of abortions due to Neosporosis, maternal serology can be a good indicator of the abortion risk in individual cows, and that the effects of factors normally related to abortion, such as parity and pregnancy season, may be masked. PMID- 15525364 TI - Canine coronavirus infection in Turkish dog population. AB - Canine coronavirus (CCoV) is one of the most important viral agents affecting the gastrointestinal system of dogs. In this study virological and serological investigations were performed to demonstrate the existence and prevalence of CCoV infection in a Turkish dog population. A total of 269 animals were subjected to the study. Of 179 dogs tested for CCoV antibodies, 112 (62.5%) were found to be positive by serum neutralization test, while 133 (74.3%) were positive by ELISA. The highest prevalence (94.2%) was detected in kennel dogs. Detection of CCoV genome in faeces was performed in samples from 90 diarrhoeic puppies by reverse transcription-polymerase chain reaction. Fourteen (15.5%) faeces were positive for CCoV RNA, five of which were characterized as CCoV type I. The widespread CCoV infection in the Turkish dog population may be attributed as an important cause of viral diarrhoea in dogs. PMID- 15525365 TI - Towards the rational use of antibiotics: results of the first International Symposium on the Risk Analysis of Antibiotic Resistance. AB - The results of four expert working groups on the risk analysis process on anti microbial resistance are described. They focus on the procedure recommended by the FAO/WHO CODEX Alimentarius Commission in 1999 including hazard identification, hazard characterization, exposure assessment and risk characterization. In all four areas, the current scientific knowledge was evaluated and recommendations for risk management options were given. They will form the basis for mitigating the health risk caused by resistant micro organisms. PMID- 15525366 TI - Veterinary use of antimicrobials and emergence of resistance in zoonotic and sentinel bacteria in the EU. AB - Antimicrobials are essential for treatment of sick animals, but even if used correctly, may eventually lead to antimicrobial resistance. While this represents a potential hazard to humans, the great majority of resistant human pathogens, especially the more important ones, are unrelated to animal sources. A survey of informed medical opinion suggested that of the human antimicrobial resistance problem, <4% was seen as potentially linked to animal sources. This proportion related largely to zoonotic bacteria which by definition have the capacity to carry resistance between species, although the evidence for resulting harm remains limited. A recent study compared resistance among chicken, pig and cattle isolates of Salmonella spp., Campylobacter spp. and Escherichia coli from a series of EU countries. When tested against antimicrobial agents, this survey showed variation of resistance between countries, between hosts and between organisms. Such variation may give insight into preferred methods of antimicrobial administration or disease control, but it is clear that the epidemiology of antimicrobial resistance induction and dissemination in animals remains complex and is yet to be fully understood. PMID- 15525367 TI - Spread of resistant bacteria and resistance genes from animals to humans--the public health consequences. AB - The paper reviews the lines of evidence which link the use of antimicrobial drugs for food animals with the emergence of antimicrobial drug resistance in bacteria pathogenic to humans, with a particular focus on the public health aspects. Deductions from the epidemiology of food-borne infections, ecological studies, outbreak investigations, typing studies and direct epidemiological observations show that resistant bacteria are transferred from food animals to man. In addition to transfer in the food chain, exchange of mobile genetic elements among commensal and pathogenic bacteria contributes to the emergence of drug resistance. There is growing evidence that this has measurable consequences for human public health. One consequence is increased transmission supported by unrelated use of anti-microbials in humans. Other consequences are related to reduced efficacy of early empirical treatment, limitations in the choices for treatment after confirmed microbiological diagnosis, and finally a possible coselection of virulence traits. Recent epidemiological studies have measured these consequences in terms of excess mortality associated with resistance, increased duration of illness, and increased risk of invasive illness or hospitalization following infections with resistant Salmonella. PMID- 15525368 TI - Glycopeptide resistant Staphylococcus. AB - Insensitivity of Staphylococcus aureus to glycopeptides became known at the end of the 1990s. To distinguish intermediate (VISA and GISA) and full resistance (vancomycin-resistant Staphylococcus aureus, VRSA) is of both epidemiological and clinical importance. The VISA (GISA) phenotype is obviously selected by glycopeptide usage in individual patients and can be disseminated by clonal spread of particularly affected staphylococcal isolates, preferentially methicillin-resistant S. aureus (MRSA). In the meantime emergence of VISA became known worldwide. VRSA evolve by acquisition of the vanA gene from enterococci, which are of significance as reservoir of resistance genes among gram-positive bacterial pathogens. Until now three independent single cases of emergence of VRSA became known from the USA, in two of them an association with vanA containing Enterococcus spp. at the same site of the affected patients could be established. The frequency of VISA (GISA) among MRSA in Central Europe is still low (<0.1%); no VRSA have been detected until now. PMID- 15525369 TI - Evidence of an association between use of anti-microbial agents in food animals and anti-microbial resistance among bacteria isolated from humans and the human health consequences of such resistance. AB - Several lines of evidence indicate that the use of anti-microbial agents in food animals is associated with anti-microbial resistance among bacteria isolated from humans. The use of anti-microbial agents in food animals is most clearly associated with anti-microbial resistance among Salmonella and Campylobacter isolated from humans, but also appears likely among enterococci, Escherichia coli and other bacteria. Evidence is also accumulating that the anti-microbial resistance among bacteria isolated from humans could be the result of using anti microbial agents in food animals and is leading to human health consequences. These human health consequences include: (i) infections that would not have otherwise occurred and (ii) increased frequency of treatment failures and increased severity of infection. Increased severity of infection includes longer duration of illness, increased frequency of bloodstream infections, increased hospitalization and increased mortality. Continued work and research efforts will provide more evidence to explain the connection between the use of anti-microbial agents in food animals and anti-microbial-resistant infections in humans. One particular focus, which would solidify this connection, is to understand the factors that dictate spread of resistance determinants, especially resistant genes. With continued efforts on the part of the medical, veterinary and public health community, such research may contribute to more precise guidelines on the use of anti-microbials in food animals. PMID- 15525370 TI - Monitoring of antimicrobial resistance among food animals: principles and limitations. AB - Large amounts of antimicrobial agents are in the production of food animals used for therapy and prophylactics of bacterial infections and in feed to promote growth. The use of antimicrobial agents causes problems in the therapy of infections through the selection for resistance among bacteria pathogenic for animals or humans. Current knowledge regarding the occurrence of antimicrobial resistance in food animals, the quantitative impact of the use of different antimicrobial agents on selection for resistance and the most appropriate treatment regimes to limit the development of resistance is incomplete. Programmes monitoring the occurrence and development of resistance are essential to determine the most important areas for intervention and to monitor the effects of interventions. When designing a monitoring programme it is important to decide on the purpose of the programme. Thus, there are major differences between programmes designed to detect changes in a national population, individual herds or groups of animals. In addition, programmes have to be designed differently according to whether the aim is to determine changes in resistance for all antimicrobial agents or only the antimicrobial agents considered most important in relation to treatment of humans. In 1995 a continuous surveillance for antimicrobial resistance among bacteria isolated from food animals was established in Denmark. Three categories of bacteria, indicator bacteria, zoonotic bacteria and animal pathogens are continuously isolated from broilers, cattle and pigs and tested for susceptibility to antimicrobial agents used for therapy and growth promotion by disc diffusion or minimal inhibitory concentration determinations. This programme will only detect changes on a national level. However, isolating the bacteria and testing for several antimicrobial agents will enable us to determine the effect of linkage of resistance. Since 1995 major differences in the consumption pattern of different antimicrobial agents have occurred in Denmark. The Danish monitoring programme has enabled us to determine the effect of these changes on the occurrence of resistance. The Danish monitoring is, however, not suited to determine changes on a herd level or to detect emergence of new types of resistance only occurring at a low level. PMID- 15525371 TI - Resistance of Salmonella isolates in Germany. AB - During 2000-2002 the National Veterinary Reference Laboratory for Salmonella (NRL Salm) in Germany typed 11,911 isolates from animals, food, feed and the environment. All of them were tested for their susceptibility to 17 anti microbial agents. Sixty-three per cent of all isolates were resistant and 40% were multiresistant (resistant against more than one anti-microbial). This general resistance level was strongly influenced by those specific serotypes which dominate the Salmonella epidemiology in Germany. Salmonella Typhimurium DT104 isolates from pig and cattle, and their resulting food products, were multiresistant in 98 and 94% of the cases respectively. During the period 2000 2003 an increasing quinolone resistance especially in Salmonella isolates from poultry and poultry meat (to 26%) and in S. Paratyphi B D-tartrate positive isolates (to 64%) could be observed. This increase was accompanied by a shift towards higher minimal inhibitory concentrations for ciprofloxacin. PMID- 15525372 TI - Hazard potential from antibiotic-resistant commensals like Enterococci. PMID- 15525373 TI - Microbial Threat--The Copenhagen Recommendations initiative of the EU. AB - The Microbial Threat was the name of an invitational EU conference held in Copenhagen in 1998, which for the first time made antibiotic resistance an official EU issue. The initiative resulted in a set of conclusions, which together with a summary of the conference was published as The Copenhagen Recommendations. This was soon followed by an EU Council Recommendation in 2001 exhorting the member states to follow and enact the recommendations from the Copenhagen meeting. Funding from EU research funds was later provided for projects monitoring resistance as well as antibiotic use, both of which have been lacking in most EU countries until now. The initiative is a good example of a major health issue being lifted by a concerted and official EU action. PMID- 15525374 TI - Conclusions and activities of previous expert groups: the Scientific Steering Committee of the EU. AB - In 1998, the EU Commission consulted its Scientific Steering Committee (SSC) to give advice on actions against anti-microbial resistance based on scientific evidence. The SSC set up a working group and adopted in 1999 an Opinion on Antimicrobial Resistance. Statements given in the well-structured document are clear, and precise recommendations were proposed. Summarizing, the Committee stated: There is evidence to suppose a continuous flow of resistance genes between pathogenic and commensal bacteria and of transfer of these bacteria between different compartments of the biosphere, thus changing the genetic resources continuously. There exist numerous factors which influence the emergence and spread of anti-bacterial resistance. However, it is likely that restriction in the use of anti-microbials will lead to a containment or a reduction of the drug resistance problem. Actions should be taken promptly to reduce the overall use of anti-microbials in a balanced way in all areas: human medicine, veterinary medicine, animal production and plant protection. PMID- 15525375 TI - Risk Analysis of Antibiotic Resistance, Berlin, BfR, 10-12 November 2003: activities of the Codex Alimentarius Commission related to antibiotic resistance. AB - The FAO/WHO Codex Alimentarius Commission develops recommendations concerning risk management related to regulatory issues, on the basis of scientific advice provided by FAO and WHO. The complex question of anti-microbial resistance that requires a multidisciplinary approach, has been considered from different point of views in the framework of Codex, especially food hygiene and veterinary drugs and to a lesser extent, plant protection. Antibiotic resistance was considered from the perspective of the food chain and a risk profile was developed. Risk management options were also considered and may be grouped in two main categories: prevention of contamination through the food chain; and prudent use of anti-microbials. It is recognized that further scientific advice is necessary before developing specific risk management options in the framework of Codex. It is expected that the results of the FAO/OIE/WHO Expert Consultations on anti microbial resistance will provide the necessary guidance for further action. PMID- 15525376 TI - OIE International Standards on Antimicrobial Resistance. PMID- 15525377 TI - Developing new regulatory approaches to antimicrobial safety. AB - Resistance to antimicrobial agents is of concern to public health officials worldwide. In industrialized countries, a significant source of antimicrobial resistant food-borne infections in humans is the acquisition of resistant bacteria originating from animals. The US Food and Drug Administration (FDA) is committed to resolving the public health impact arising from the use of antimicrobial drugs in food-producing animals. The FDAs goal is to ensure that significant human antimicrobial therapies are not compromised or lost while providing for the safe use of antimicrobials in food animals. Recently the FDA published a guidance document titled 'Evaluating the Safety of Antimicrobial New Animal Drugs with Regard to their Microbiological Effects on Bacteria of Human Health Concern' (US Food and Drug Administration, Center for Veterinary Medicine, 2003). This document outlines a pathway drug sponsors can use to address concerns about antimicrobial resistance prior to approval of their drug. The process uses a qualitative risk assessment approach to assess the potential of the intended use of a product to develop resistance in bacteria that may harm humans. The level of risk determines the level of risk management that is required for the drug to be used. The Food and Drug Administration (FDA) always has the option of not approving a drug if the risk of a public health consequence is too high. PMID- 15525378 TI - Antimicrobial resistance in bacteria from food-producing animals. risk management tools and strategies. AB - The application of antimicrobial agents has proved to be the main risk factor for development, selection and spread of antimicrobial resistance. This link applies to the use of antimicrobial agents in human and in veterinary medicine. Furthermore, antimicrobial-resistant bacteria and resistant genes can be transmitted from animals to humans either by direct contact or via the food chain. In this context, risk management has to be discussed regarding prevention and control of the already existing antimicrobial resistance. One of the primary risk management measures in order to control the development and spread of antimicrobial resistances is by regulating the use of antimicrobial agents and subjecting their use to guidelines. Thereby, the occurrence of antimicrobial resistant bacteria in the human and veterinary habitat can be controlled to a certain degree. There is little information about past attempts to prevent the development of resistances or to control them, and even less is known about the effectiveness or the cost intensiveness of such efforts. Most of the strategies focus on preventing and controlling antimicrobial resistance by means of the reduction or limitation of the use of antimicrobial agents in food-producing animals. PMID- 15525380 TI - Initial biofilm formation of Streptococcus sobrinus on various orthodontics appliances. AB - Biofilms accumulate on hard and soft surface in the oral cavity. Accumulation of biofilms on orthodontic appliance bear scientific and clinical interest. The objection of this study was to examine the formation of dental biofilm by Streptococcus sobrinus on different types of orthodontics appliances, using a model consisting of host and bacterial constituents. The adsorption pattern of saliva to the orthodontics appliances was determined by means of gel electrophoresis coupled with computerized densitometry techniques. The amount of salivary proteins adsorbed onto the surfaces was measured using the Bradford method. Sucrose-dependent bacterial adhesion to the saliva-coated orthodontics appliances was tested by radioactive-labelled S. sobrinus. Our results show different adsorption patterns of salivary proteins to the various orthodontic appliances as modules, brackets, springs and intra oral elastics. Modules and brackets demonstrated the most affinity to salivary proteins. A surface dependent adhesion profile was recorded, showing a high affinity of albumin and amylase to modules. Bacterial accumulation was the highest on modules compared with springs which demonstrated the least bacterial adhesion. Our study demonstrates the specificity of biofilm formation on the different orthodontic appliances. Formation of a variety of dental biofilms has a significant impact on the progression of dental diseases associated with orthodontic treatment. PMID- 15525381 TI - The effect of Coca-Cola and fruit juices on the surface hardness of glass ionomers and 'compomers'. AB - The interaction of tooth-coloured dental restorative materials (a conventional glass-ionomer, two resin-modified glass-ionomers and two compomers) with acidic beverages has been studied with the aim of investigating how long-term contact affects solution pH and specimen surface hardness. For each material (ChemFil Superior, ChemFlex, Vitremer Core Build-Up/Restorative, Fuji II LC, Dyract AP and F2000) disc-shaped specimens were prepared and stored in sets of six in the following storage media: 0.9% NaCl (control), Coca-Cola, apple juice and orange juice. After time intervals of 1 day, 1 week, 1 month, 3 months, 4 months, 6 months and 1 year, solution pH and Vickers Hardness Number were determined for each individual specimen. Differences were analysed by anova followed by Student Newman-Keuls post hoc analysis. All materials were found to reduce the pH of the 0.9% NaCl, but to increase the pH of the acidic beverages. The conventional glass ionomers dissolved completely in apple juice and orange juice, but survived in Coca-Cola, albeit with a significantly reduced hardness after 1 year. The other materials survived in apple juice and orange juice, but showed greater reductions in surface hardness in these beverages than in Coca-Cola. Fruit juices were thus shown to pose a greater erosive threat to tooth coloured materials than Coca Cola, a finding which is similar to those concerning dentine and enamel towards these drinks. PMID- 15525382 TI - An in vitro investigation of penetration depth of dentine bonding agents into carious dentine. AB - The aim of the present in vitro study was to determine the penetration depth of dentine bonding agents into carious dentine as a possible strategy in caries treatment of deep lesions. Forty-eight extracted carious teeth were used for the experiments and divided in four groups. In a split-tooth design, one half of each tooth was treated without caries excavation, and the second half after caries excavation. In group 1 the teeth were treated with Prime & Bond 2.0 without phosphoric acid etching, and in group 2 with phosphoric acid etching. In group 3, the teeth were treated with All-Bond 2 without phosphoric acid etching, and in group 4 with phosphoric acid etching. Bonding agents were marked with an orange dye prior to application. The composition of the bonding agents had no significant influence on penetration depths. The mean thickness of the outer, necrotic caries zone in unexcavated specimens was 117 microm (+/-126). Bonding agents penetrated significantly deeper into unexcavated carious dentine (mean 124 +/- 76 microm) compared with excavated dentine (11 +/- 14 microm) and sound dentine (3 +/- 4 microm). Acid etching significantly reduced the thickness of the outer caries zone, and the penetration depth of the bonding agents into unexcavated carious specimens by approximately 50%, and increased it into sound dentine. The results indicate that dentine bonding agents could completely penetrate the unexcavated outer carious dentine in 90% of specimens. Further studies are required to determine whether further caries activity is arrested after treatment with dentine bonding procedures. PMID- 15525383 TI - Effect of 2-hydroxyethyl methacrylate pre-treatment on micro-tensile bond strength of resin composite to demineralized dentin. AB - The purpose of this study was to evaluate the effect of 2-hydroxyethyl methacrylate (HEMA) application on the micro-tensile bond strength of resin composite to demineralized dentin. Artificially demineralized lesions were formed on bovine dentin surfaces and treated with 10, 30, 50, 70 and 100 wt% HEMA aqueous solution. The surfaces were then applied and covered with SE Bond and AP X according to the manufacturer's instruction. After immersion in 37 degrees C water for 24 h, bond strength were measured using a universal testing machine. Bond strengths to both demineralized dentin and normal dentin, without HEMA application, were also measured. Scanning electron microscopic (SEM) observation and confocal laser scanning microscopy (CLSM) analysis at the resin-dentin interface were also performed. The bond strength data were statistically compared with anova and Scheffe's test (P < 0.05). Bond strength to demineralized dentin treated with over 30 wt% HEMA aqueous solution were significantly higher than that to demineralized dentin without HEMA application, but significantly lower than that to normal dentin. SEM observation revealed that the hybrid layer and resin-tags thickened and lengthened with HEMA application. In CLSM, the diffusion of adhesive primer into demineralized dentin increased with HEMA application. These results indicated that HEMA application might increase the bond strength to demineralized dentin by the enhancement of resin monomer penetration of HEMA. PMID- 15525384 TI - The effect of pulse activation on microleakage of a 'packable' composite resin and two 'ormocers'. AB - The purpose of the study was to examine the effect of pulse polymerization on microleakage of one packable composite resin and two organically modified ceramics (ormocers), within a high C-factor preparation. Class-V cavity preparations with occlusal margins in enamel and gingival margins in dentin were prepared on 60 freshly extracted premolars or molars. Teeth were randomly assigned to 12 experimental groups (five teeth each) representing the different restorative systems, mode of polymerization and thermal cycling test. Three groups of 20 teeth each were restored with one of the following restorative systems: Scotch bond 1/Filtek P 60, Admira bond/Admira, Etch&Prime 3.0/Definite. 10 restorations of each restorative system (two experimental groups) were conventionally polymerized, whereas the other 10 restorations were polymerized with a pulse program. The specimens were stored in double-distilled water at 37 degrees C for 24 h. Ten restorations from each restorative system (30 restorations, half of the specimens) were thermocycled, the other 30 restorations were kept in double-distilled water at 37 degrees C for 24 h. Then all 60 restorations stained with dye, sectioned and scored for microleakage. Results showed that there were no significant differences (P > 0.05) among the materials with respect to microleakage scores. The experimental groups that were conventionally polymerized revealed significantly more microleakage (P < 0.05) after thermal cycling than all the other groups. The gingival microleakage was significantly more (P < 0.05) than occlusal in conventionally polymerized, thermocycled experimental groups. PMID- 15525385 TI - Surface roughness of acrylic resins submitted to mechanical and chemical polishing. AB - Surface roughness is extremely important as it influences micro-organism accumulation and consequently oral health. The aim of the present study was to evaluate the influence of mechanical polishing (MP) and chemical polishing (CP) on surface roughness of four heat-cured denture base acrylic resins. Twenty sample discs were prepared for each of the following acrylic resins: Classico (CL), QC 20 (QC), Acron MC (AC) and Onda Cryl (ON). The first two are boiling cured resins and the latter two are microwave cured. After curing and deflasking, samples were ground and subdivided into two groups: MP, which received pumice slurry, chalk powder, soft brush and felt cone, at a bench vice; and CP, which received fluid specifically for chemical polisher. Surface roughness was measured using a Surf Corder SE 1700 rugosimeter. Data collected were statistically analysed by analysis of variance followed by Tukey's test. Surface roughness means in mum were: CL-MP: 0.0350; CL-CP: 0.2298; QC-MP: 0.0307; QC-CP: 0.1792; AC MP: 0.0312; AC-CP: 0.2700; ON-MP: 0.0321; ON-CP: 0.2949. It can be concluded that MP produced significantly smoother surfaces than CP and that surface roughness because of MP was not influenced by acrylic resin type, whereas this was not true of CP. PMID- 15525386 TI - Comparison of the tensile bond strengths of cast metal crowns luted with resin cements. AB - The limitation of photoactivation of dual-polymerized resin cements along the margins of metal restorations may adversely affect the mechanical properties of these cements, thus impairing the retention of restorations. The aim of this study was to assess the bond strength of cast metal crowns cemented with three dual-polymerized resin cements, using a chemically-activated resin cement and zinc phosphate as controls. Fifty nickel-chromium alloy crowns were cast and randomly assigned to five groups of equal size. Castings were cemented on their corresponding metal dies with one of the tested luting agents: Scotchbond Resin Cement, Enforce and Panavia F (dual-polymerized resin cements), Cement-It (chemically-activated resin cement) and Zinc Phosphate Cement (zinc phosphate cement). Specimens were stored in distilled water at 37 degrees C for 24 h and then loaded in tension until failure. Panavia F and Zinc Phosphate Cement provided the highest and lowest bond strength means, respectively. Scotchbond Resin Cement, Enforce and Cement-It cements exhibited similar intermediate values, but with statistically significant difference compared to the other materials (P < 0.05). Even with the restriction or absence of light activation, all tested dual-polymerized resin cements produced significantly higher bond strength than did the zinc phosphate cement and yielded similar or better results than the chemically activated cement. It should be pointed out that the findings of this study relate to a test scenario which does not mimic clinical circumstances and that further work is required to identify the clinical significance of the reported tensile bond strength differences between the different luting materials. PMID- 15525387 TI - Hardness and compressive strength of indirect composite resins: effects of immersion in distilled water. AB - The aim of this study was to evaluate the effect of ageing in distilled water on the hardness and compressive strength of a direct composite resin Z100, a feldspatic porcelain (Noritake) and three indirect composites (Artglass, Solidex and Targis). For the Vickers hardness tests, five disk-shaped specimens (2 x 4 mm) of each material were prepared according to the manufacturers' instructions. The hardness tests were conducted using a Vickers diamond indentor. Compressive strength measurements were recorded on cylindrical specimens with a diameter of 6 mm and a length of 12 mm. The compression tests were carried out with a constant cross-head speed of 0.5 mm min(-1) on a mechanical test machine. For each material, 10 specimens were tested after 7 days of dry storage at 37 +/- 1 degrees C and 10 specimens were tested after water storage at 37 +/- 1 degrees C for 180 days. Noritake porcelain specimens showed higher hardness values than the composites. Among the composite materials, Z100 promoted the highest VHN values, regardless of the ageing periods. The results showed that Solidex and Z100 had the highest compressive strength values. Ageing in water reduced the hardness for all composites, but had no long-term effect on the compressive strength. PMID- 15525389 TI - Calculation of colour resulting from composite/compomer layering techniques. AB - This study determined the influence of optical properties of constituent layers on the colour of double-layer aesthetic filling materials. Multiple regression equations for the Commission Internationale de I'Eclairage (CIE) L*, a* and b* of layered materials were calculated from the optical values of the covering and underlying layers. Specimens (10 mm diameter, 1 mm thickness) of two light-cured resin composites and one compomer of seven to 11 shades were used. CIE L*, a* and b* values of each specimen were measured with a colour spectrophotometer backed by a standard white background. The scattering coefficient (S), absorption coefficient (K), contrast ratio (C) and translucency parameter (T) were calculated. Double-layered specimens were formed in optical contact by joining two different shades from the same material, or resin composite as covering with a compomer underlying layer. Each of the L*, a* and b* of layered material was used as a dependent variable, and 14 optical values of underlying and covering layers were used as independent variables in forward regression analysis (P = 0.01). CIE L* after layering had a positive correlation with S of covering layer (correlation coefficient; beta = 0.79-0.91, P < 0.01) and a correlation with L* of underlying layer (beta = 0.14-0.16). CIE a* after layering had a correlation with a* of covering layer (beta = 0.83-0.94) and a correlation with a* of underlying layer (beta = 0.30-0.56). CIE b* after layering had a correlation with b* of covering layer (beta = 0.77-0.90) and a correlation with T of covering layer (beta = 0.40-0.59). The layered colour of these materials can be predicted by the derived regression equations within the limitations of this study. CIE L*, a* and b* values of double-layer material are mainly influenced by S, CIE a* and b* of covering layer, respectively. PMID- 15525388 TI - Characterization of the inorganic fraction of resin composites. AB - The purposes of this study were to determine the weight fraction of filler in thirty-nine resin-based materials including flowable and packable composites, and to examine the morphology of the filler particles. The percentages of inorganic fillers by weight were determined by Thermogravimetric Analysis and by ashing in air technique at 900 degrees C. The size and shape of the filler particles were examined using scanning electron microscopy (SEM) after dissolution of the organic matrix. The weight fraction of inorganic fillers ranged between 41.6 and 84.6%. Wide variations were found among materials of the same category. Values found in the present study were sometimes different from those given by the manufacturers. The SEM photomicrographs showed various shapes, and sizes of inorganic fillers. Compared with universal hybrid restorative materials, flowable composites have lower filler loading and packable resin composites did not show higher values as claimed by some manufacturers. Various factors may explain the observed discrepancies between the manufacturer's data and our results. The silane treatment as well as the incorporation of organic material as part of the fillers of the composite could be responsible for those differences. PMID- 15525390 TI - Mechanical properties of cast Ti-6Al-4V-XCu alloys. AB - The mechanical properties of Ti-6Al-4V-XCu (1, 4 and 10 wt% Cu) alloys were examined. The castings for each alloy were made in a centrifugal titanium casting machine. Two shapes of specimens were used: a dumbbell (20 mm gauge length x 2.8 mm diameter) for mechanical property studies, and a flat slab (2 mm x 10 mm x 10 mm) for metallography, microhardness determination and X-ray diffractometry. Tensile strength, yield strength, modulus of elasticity, elongation and microhardness were evaluated. After tensile testing, the fracture surfaces were observed using scanning electron microscopy. The tensile strengths of the quaternary alloys decreased from 1016 MPa for the 1% Cu alloy to 387 MPa for the 10% Cu alloy. Elongation decreased with an increase in the copper content. The 1% Cu alloy exhibited elongation similar to Ti-6Al-4V without copper (3.0%). The results also indicated that the copper additions increased the bulk hardness of the quaternary alloy. In particular, the 10% Cu alloy had the highest hardness and underwent the most brittle fracture. The mechanical properties of cast Ti-6Al 4V alloy with 1 and 4% Cu were well within the values for existing dental casting non-precious alloys. PMID- 15525391 TI - Physical properties and compatibility with dental stones of current alginate impression materials. AB - This study examined physical properties and compatibility with dental stones of two types of alginate impression materials. Five powder-type alginate impression materials (Alginoplast EM, Aroma Fine, Algiace Z, Coe Alginate, Jeltrate Plus) and a paste-type alginate impression material (Tokuso AP-1) were used. The dynamic viscosity immediately after mixing was measured by means of a controlled stress rheometer. The gelation times were determined according to Japanese Industrial Standards (JIS) T6505, and recovery from deformation, strain in compression and compressive strength were determined according to the International Organization for Standardization (ISO) specification 1563. Detail reproduction and surface roughness of type III dental stones (New Plastone, New Sunstone) and a type IV dental stone (Die Stone) were evaluated using a ruled test block as specified in the ISO specification 1563 and a profilometer, respectively. The alginate impression materials evaluated in this study were all in compliance with the ISO specification 1563 and JIS T6505. The alginate impression materials had similar mechanical properties after gelation, whilst a wide range of dynamic viscosity immediately after being mixed, gelation times and compatibility with dental stones were found among the materials. The paste-type material had a higher dynamic viscosity and a shorter gelation time than the powder-type materials. The best surface quality was obtained with the paste-type material/type III dental stone cast combinations. The materials should be selected in consideration of initial flow, setting characteristics and compatibility with dental stones. The results suggested that a paste-type material would better meet the requirements of an alginate impression material. PMID- 15525392 TI - The effect of retainer thickness on posterior resin-banded prostheses: a finite element study. AB - According to its design concept, a resin-bonded prosthesis, compared with the conventional fixed partial denture, is a weak and unstable structure. Therefore, a resin-bonded prosthesis induces a higher failure rate, especially in the posterior region. Recently, adhesion agents have been profoundly improved. However, the design guidelines of posterior resin-bonded prostheses (RBP) have seldom been evaluated from a biomechanical perspective. The objective of this study was to investigate the biomechanical effects of the retainer thickness on posterior RBP using the finite element method. A solid model of a posterior mandibular resin-bonded prosthesis, which employed the second molar and second premolar as the abutment teeth, was constructed and meshed with various retainer thickness (ranging from 0.2 to 1.0 mm). Horizontal and vertical loadings of 200 N were applied respectively at the central fossa of the pontic to examine the stress level at the interface between the retainer and abutment teeth. All exterior nodes in the root, below the cementoenamel junction were fixed as the boundary condition. The results showed that horizontal loading would induce higher interfacial stresses than the vertical loading which indicated that the horizontal component of the occlusal force plays a more important role in evaluating the debonding phenomenon. Further, the peak interfacial stresses increased as the retainer thickness decreased and, based on the fitted relation between retainer thickness and interfacial stresses, a 0.4 mm retainer thickness was suggested as the minimum required to prevent severe interfacial stresses increasing. PMID- 15525393 TI - Efficacy of conventional and experimental techniques for denture repair. AB - This study investigated the efficacy of one experimental and three conventional techniques for denture repair. Forty maxillary dentures were constructed in dental stone casts duplicated from an edentulous copper-aluminium maxillary master cast. Two groups of 20 dentures each were prepared with Lucitone 199 (water-bath, 8 h/74 degrees C) or Acron MC (microwave, 3 min/500 W) denture base materials processed in gypsum moulds. The 40 dentures were all separated sagittally in the middle. After that, five dentures of each denture base material were repaired with one of the four following techniques: L (Lucitone 199, water bath, 8 h/74 degrees C, gypsum mould), A (Acron MC, microwave, 3 min/500 W, gypsum mould), AR (Acron MC/R, autopolymerized, 60 psi/45 degrees C/15 min) and the experimental technique AS (Acron MC, 1 min/500 W + 1 min/0 W + 1 min/500 W, hard silicone mould). The parameters denture accuracy (DA), horizontal (HC) and vertical changes (VC) of the occlusal plane measured the efficacy of the repair techniques. The DA was determined by weighing a film of silicone impression material set in contact to the tissue surface of the denture seated on the metallic master die. For HC, cross-arch measurements were made among reference marks drilled on the teeth 11, 21, 16 and 26. The VC was obtained by calculating the relative differences in height between similar teeth of each semi-arch (pairs 13-23, 14-24, 15-25 e 16-26). For DA, HC and VC, the percentage differences between the percentage means obtained before and after repair were calculated and grouped for comparisons. Analysis of variance (SuperANOVA) and means compared by Tukey-Kramer intervals (0.05) revealed that AR repair had the best percentage difference value for DA [0.5% (P < 0.05)], while the others were not statistically different [L = 27.2%, A = 28.9%, AS = 21.2% (P > 0.05)]. For HC, there was a statistical difference (P < 0.05) between AR and the other techniques for the tooth pairs, 16-26, 11-26 and 21-16; repairs with AR and AS differed for the 11-21 pair, while those with A and AS techniques differed for the 16-26 pair (P < 0.05). The VC differences were not detected between repair methods (P > 0.05). Denture accuracy was not affected by the interaction of base material repair technique; repair with AR technique gave the best adaptation; the interaction of base material-repair technique did not affect HC; HC was affected by the repair technique. PMID- 15525394 TI - Shear modulus and thermal properties of gutta percha for root canal filling. AB - The purpose of this investigation was to investigate the rheological properties of four commercially available gutta perchas for root canal filling. The relaxation modulus [Gr(0): instantaneous shear modulus] and specific volume of their materials were examined. In addition, the quantity of heat was also studied by differential scanning calorimeter. In a lower temperature range than the first order transition temperature (melting point), the Gr(0) values of each material were almost identical. A marked decrease of Gr(0) was observed at the melting point, and the range of the first-order transition temperature at heating was from 42.0 to 60.0 degrees C. At higher temperatures than the first-order transition temperature of each material, a considerable difference in Gr(0) values was observed. The transition temperatures obtained by the results of the Gr(0), specific volume and quantity of heat agreed with one another. A marked specific volume change was observed at the first-order transition temperature. The technique using melted gutta percha may not be favourable compared with the conventional lateral condensation technique because melted gutta percha undergoes a large amount of shrinkage during setting. PMID- 15525395 TI - Effect of the gutta-percha solvents on the microhardness and the roughness of human root dentine. AB - The purpose of this in vitro study was to evaluate the effect of gutta-percha solvents on the microhardness and the roughness of human root dentine. Forty-five recently extracted human mandibular anterior teeth were used. The teeth were sectioned longitudinally, embedded in acrylic resin and polished. Ninety specimens were divided into three groups containing 30 teeth each. In group 1, the specimens were treated with chloroform for 15 min. In group 2, the specimens were treated with halothane for 15 min. In group 3, distilled water was used and this group was served as control. All groups were then divided into two subgroups of 15 specimens each. The specimens, in groups 1a, 2a and 3a, were submitted to Vicker's microhardness indentations. Groups 1b, 2b and 3b were used for determination of the roughness of root dentine. The data were recorded as Vicker's numbers and Ra, mum for roughness test. The results were then analysed by one-way ANOVA and post hoc Tukey tests. The results indicated that chloroform and halothane did not affect the microhardness and the roughness of root dentine (P > 0.05). PMID- 15525396 TI - Costs and benefits of genetic heterogeneity within organisms. AB - An increasing number of studies have recently detected within-organism genetic heterogeneity suggesting that genetically homogeneous organisms may be rare. In this review, we examine the potential costs and benefits of such intraorganismal genetic heterogeneity (IGH) on the fitness of the individual. The costs of IGH include cancerous growth, parasitism, competitive interactions and developmental instability, all of which threaten the integrity of the individual while the potential benefits are increased genetic variability, size-specific processes, and synergistic interactions between genetic variants. The particular cost or benefit of IGH in a specific case depends on the organism type and the origin of the IGH. While mosaicism easily arise by genetic changes in an individual, and will be the more common type of IGH, chimerism originates by the fusion of genetically distinct entities, and is expected to be substantially rare in most organisms. Potential conflicts and synergistic effects between different genetic lineages within an individual provide an interesting example for theoretical and empirical studies of multilevel selection. PMID- 15525397 TI - Will two walk together, except they have agreed? Amos 3:3. PMID- 15525398 TI - Intraorganismal genetic heterogeneity: is it a useful concept? PMID- 15525399 TI - Genetic heterogeneity within organisms and the evolution of individuality. PMID- 15525400 TI - Much ado about nothing... so far? PMID- 15525401 TI - Mosaicism and chimerism as components of intraorganismal genetic heterogeneity. PMID- 15525402 TI - Genetic conflicts and intercellular heterogeneity. PMID- 15525404 TI - Spite and the scale of competition. AB - In recent years there has been a large body of theoretical work examining how local competition can reduce and even remove selection for altruism between relatives. However, it is less well appreciated that local competition favours selection for spite, the relatively neglected ugly sister of altruism. Here, we use extensions of social evolution theory that were formulated to deal with the consequences for altruism of competition between social partners, to illustrate several points on the evolution of spite. Specifically, we show that: (i) the conditions for the evolution of spite are less restrictive than previously assumed; (ii) previous models which have demonstrated selection for spite often implicitly assumed local competition; (iii) the scale of competition must be allowed for when distinguishing different forms of spite (Hamiltonian vs. Wilsonian); (iv) local competition can enhance the spread of spiteful greenbeards; and (v) the theory makes testable predictions for how the extent of spite should vary dependent upon population structure and average relatedness. PMID- 15525405 TI - A phylogenomic approach to reconstructing the diversification of serine proteases in fungi. AB - Using a phylogenomic approach with 10 fungi of very different virulence and habitat, we determined that there was substantial diversification of subtilase type proteases early in ascomycete history (with subsequent loss in many lineages) but with no comparable diversification of trypsins. Patterns of intron loss and the degree of divergence between paralogues demonstrated that the proliferation of proteinase K subtilases and subtilisin type subtilases seen in pathogenic ascomycetes (Metarhizium anisopliae, Magnaporthe grisea, Fusarium graminearum) occurred after the basidiomycete/ascomycete split but predated radiation of ascomycete lineages. This suggests that the early ascomycetes had a lifestyle that selected for multiple proteases, whereas the current disparity in gene numbers between ascomycete lineages results from retention of genes in at least some pathogens that have been lost in other lineages (yeasts, Aspergillus nidulans, Neurospora crassa). A similar prevailing trend towards lineage specific gene loss of trypsins in saprophytes and some pathogens suggests that their phylogenetic breadth will have been much wider in early fungi than currently. PMID- 15525406 TI - Offspring-driven local dispersal in female sand lizards (Lacerta agilis). AB - We report on a field study in which determinants of female breeding dispersal (i.e. the shift in the mean home range coordinates between successive breeding events) was investigated. Offspring were released in full sib groups (or half sib ones if there was within-clutch multiple paternity) at a separation distance from the females that varied between 'families'. This allowed for analysis of 'offspring nearness' effects on maternal dispersal. When a female's offspring were released more closely to her, she responded with greater dispersal. Furthermore, when the data set was truncated at 100 m maternal-offspring separation distance at offspring release (because perception at longer distances is likely to be unrealistic), maternal dispersal resulted in greater separation distance between female and offspring in the following year. A corresponding analysis for juveniles revealed no effect of maternal nearness on offspring dispersal but identified a significant effect of clutch size, to our surprise with dispersal declining with increasing clutch size. We discuss this result in a context of the 'public information hypothesis' (reinterpreted for juveniles in a nonsocial foraging species), suggesting that conspecific abundance perhaps acts as an indicator of local habitat quality. Thus, our analysis suggests a microgeographic structuring of the adult female population driven by genetic factors, either through inbreeding avoidance, or from simply avoiding individuals with a similar genotype regardless of their pedigree relatedness, while a nongenetic factor seems more important in their offspring. PMID- 15525407 TI - The close relationship between estimated divergent selection and observed differentiation supports the selective origin of a marine snail hybrid zone. AB - To study the role of divergent selection in the differentiation of the two morphs in a hybrid zone of the intertidal snail Littorina saxatilis, we compared the strength of the divergent selection acting on a series of shell characters (as estimated by the viability of snails in a reciprocal transplant experiment) with the contribution of these characters to the phenotypic differences between the morphs. We found a close correlation between selection and differentiation, which suggests a cause-effect relationship, i.e. that all present differentiation is the result of past divergent selection. In addition, divergent selection was a very important component of the total natural selection acting on shell measures. These novel results support previous evidence, based on allozyme analysis, of a parapatric origin for this hybrid zone. We discuss possible limitations of this interpretation and the circumstances under which allopatric differentiation would produce the same results. Phenotypic analysis of divergent selection may be a useful method of investigating the evolutionary mechanisms involved in differentiation processes. PMID- 15525408 TI - Evolution of viviparity in horned lizards (Phrynosoma): testing the cold-climate hypothesis. AB - The cold-climate hypothesis for evolution of viviparity in squamates predicts a correlation between reproductive mode, altitude and latitude. I tested this prediction in horned lizards within a phylogenetic context. I first determined whether all viviparous species were monophyletic using Monte Carlo simulations. Secondly, I tested for presence of phylogenetic signal using randomization tests. Thirdly, I analysed relationships between reproductive mode and minimum, midpoint, and maximum altitudes and latitudes by computing conventional correlations and phylogenetically independent contrasts. Viviparous species do not form a monophyletic group suggesting viviparity evolved twice in the genus. Viviparity and altitude showed strong phylogenetic signal based on randomization tests and were significantly correlated, while latitude was not correlated with reproductive mode. This study partially supports the cold-climate model, but also suggests that altitude either may be a better predictor of cold temperatures or may be a surrogate for other selective factors important in the evolution of viviparity. PMID- 15525409 TI - Sexual isolation and extreme morphological divergence in the Cumana guppy: a possible case of incipient speciation. AB - Theory predicts that sexual selection can promote the evolution of reproductive isolation and speciation. Those cases in which sexual selection has led to speciation should be characterized by significant differentiation in male display traits and correlated female preferences in the absence of post-zygotic isolation, accompanied by little genetic or other morphological differentiation. Previous evidence indicates that a cluster of populations of the guppy (Poecilia reticulata Peters) from Cumana, Venezuela, the 'Cumana guppy', differs significantly in female preferences from a nearby guppy population (A. Lindholm & F. Breden, Am. Nat., 160: 2002, S214). Here, we further document sexual isolation between these populations. In addition, these populations exhibit significant divergence in male display traits correlated to differences in between-population mating success, little mitochondrial genetic differentiation, and we find no evidence for genetic incompatibility between a Cumana population and several geographically isolated populations. These results suggest that divergent sexual selection has contributed to differentiation of the Cumana guppy, and this may be the first example of incipient speciation in the guppy. PMID- 15525410 TI - The effect of temperature and wing morphology on quantitative genetic variation in the cricket Gryllus firmus, with an appendix examining the statistical properties of the Jackknife-MANOVA method of matrix comparison. AB - We investigated the effect of temperature and wing morphology on the quantitative genetic variances and covariances of five size-related traits in the sand cricket, Gryllus firmus. Micropterous and macropterous crickets were reared in the laboratory at 24, 28 and 32 degrees C. Quantitative genetic parameters were estimated using a nested full-sib family design, and (co)variance matrices were compared using the T method, Flury hierarchy and Jackknife-manova method. The results revealed that the mean phenotypic value of each trait varied significantly among temperatures and wing morphs, but temperature reaction norms were not similar across all traits. Micropterous individuals were always smaller than macropterous individuals while expressing more phenotypic variation, a finding discussed in terms of canalization and life-history trade-offs. We observed little variation between the matrices of among-family (co)variation corresponding to each combination of temperature and wing morphology, with only one matrix of six differing in structure from the others. The implications of this result are discussed with respect to the prediction of evolutionary trajectories. PMID- 15525411 TI - Genetic and morphological data supporting the hypothesis of adaptive radiation in the endemic fish of Lake Matano. AB - Adaptive radiation resulting from differential selection acting on functional features is believed to be an important source of biodiversity. In this study, morphometric measures and mitochondrial DNA are used to test for adaptive radiation within four fish genera (Glossogobius, Oryzias, Dermogenys and Telmatherina) endemic to an ancient island lake (Lake Matano, Sulawesi, Indonesia), using the framework proposed by Schluter (The Ecology of Adaptive Radiation, Oxford University Press, 2000). We demonstrate common ancestry and rapid divergence in one genus (Telmatherina) based on 560 bp of 16S sequence data. We found higher levels of variation in feeding-related traits (N = 8) for Telmatherina relative to the other genera, while no differences were found for sexual display traits (N = 8) or neutral morphological traits (N = 8). Telmatherina also had the highest number of distinct colouration patterns among the four genera. These data, combined with the very low productivity of the lake, are indicative of selection driving adaptive radiation. The morphometric divergence in the Telmatherina likely results from selection acting on feeding traits in this low productivity lake, leading to trophic specialization among closely related morphotypes. These results provide indirect but compelling data supporting the adaptive radiation of Telmatherina in this system. PMID- 15525412 TI - Quantitative genetics of sexually dimorphic traits and capture of genetic variance by a sexually-selected condition-dependent ornament in red junglefowl (Gallus gallus). AB - We studied the quantitative genetics of sexually selected traits in a captive population of red junglefowl (Gallus gallus L.) using a multi-generational 'animal model' approach. We found significant heritability of mass, tarsus length (both strongly sexually dimorphic), residual mass, and male comb (a fleshy head ornament) length. Residual mass has a genetic correlation between the sexes smaller than unity and so could show partially independent responses to selection in the two sexes. In males, tarsus length and mass were not genetically correlated, and this produced a negative genetic correlation between tarsus length and residual mass. The male red junglefowl's comb, an ornament influencing female choice, is highly condition dependent. We show that expression of this ornament is heritable, however, and shows strong genetic correlation with a condition index, residual mass. Because residual mass is partly influenced by various aspects of condition, it appears that comb size has 'captured' genetic variability in condition. PMID- 15525413 TI - Genetic variation in carbon isotope discrimination in six European populations of Castanea sativa Mill. originating from contrasting localities. AB - The objective of this study was to evaluate the variability of physiological performances of Castanea sativa Mill. in relation to drought tolerance, among and within European populations coming from contrasting environmental conditions. Forty-eight open-pollinated families from a stratified sample (temperature/precipitation) of six naturalized populations from Spain, Italy and Greece were grown for one growth period under two temperature regimes (25 and 32 degrees C), in combination with two watering regimes in growth chambers. Complementary to growth traits analysed in a previous study, carbon isotope discrimination (Delta), a complex physiological trait involved in acclimation and adaptive processes, was studied. anova indicated significant Delta variability for C. sativa populations across Europe and, thereby, variation in adaptedness to drought. The European pattern of Delta variability matches the previously reported one for the centre of origin of C. sativa (Ponto-Caucasian region). This suggests that common mechanisms of drought adaptedness, involving both genetic and physiological determinants, give C. sativa the capacity to colonize a wide range of site conditions. The highest Delta values, indicating the lowest water use efficiency (WUE), were found within each treatment for populations originating from Mediterranean drought-prone sites. These populations also had the highest phenotypic plasticity of Delta. Significant among-family genetic variation in Delta was found. The heritability based on the joint anova was estimated at 0.31 +/- 0.07. The estimates of the coefficients for the additive variance varied in the range 2.6-4.0%, suggesting possibilities for selection on WUE and adaptedness to drought. The genetic correlations between Delta and growth traits were generally strong and negative, especially in the two high temperature treatments. PMID- 15525414 TI - How do natural and sexual selection contribute to sympatric speciation? AB - I use explicit genetic models to investigate the importance of natural and sexual selection during sympatric speciation and to sort out how genetic architecture influences these processes. Assortative mating alone can lead to speciation, but rare phenotypes' disadvantage in finding mates and intermediate phenotypes' advantage due to stabilizing selection strongly impede speciation. Any increase in the number of loci also decreases the likelihood of speciation. Sympatric speciation is then harder to achieve than previously demonstrated by many theoretical studies which assume no mating disadvantage for rare phenotypes and consider a small number of loci. However, when a high level of assortative mating evolves, sexual selection might allow populations to split into dimorphic distributions with peaks corresponding to nearly extreme phenotypes. Competition then works against speciation by favouring intermediate phenotypes and preventing further divergence. The interplay between natural and sexual selection during speciation is then more complex than previously explained. PMID- 15525415 TI - Heightened condition dependence is not a general feature of male eyespan in stalk eyed flies (Diptera: Diopsidae). AB - Stalk-eyed flies are exemplars of sexual selection leading to the evolution of exaggerated male ornaments (eyespan). In Sphyracephala beccarri, there is no evidence for female mate choice for exaggerated male eyespan and only minor sex differences in eyespan. We used S. beccarri to test whether heightened condition dependence only evolves when male eyespan becomes sexually exaggerated. Male eyespan showed heightened condition dependence under food stress compared with a control trait (wing length). However, female eyespan displayed a similar pattern and there was no sex difference in the degree of increased eyespan sensitivity. The finding that eyespan is a sensitive indicator of food stress, even in an unexaggerated state, suggests that this may have acted as a pre-adaptation to the role of eyespan in sexual signalling in other Diopsid species. These results are consistent with handicap theory and Fisher's view of how sexual selection is initiated. PMID- 15525416 TI - Complexity and integration in sexual ornamentation: an example with carotenoid and melanin plumage pigmentation. AB - Sexual ornaments often consist of several components produced by distinct developmental processes. The complexity of sexual ornaments might be favoured by mate choice of individual components in different environments which ultimately results in weak interrelationships (integration) among the developmental processes that produce these components. At the same time, sexual selection for greater exaggeration of individual components favours their stronger co dependence on organismal resources. This should ultimately produce stronger condition-mediated integration among ornaments' components in individuals with the most exaggerated ornamentation. Here we distinguish between these two sources of integration by examining the relationship between integration and elaboration of sexual ornamentation in three bird species: two with carotenoid-based sexual ornamentation (the house finch, Carpodacus mexicanus and common redpoll, Carduelis flammea) and a species with melanin-based sexual ornamentation (house sparrow, Passer domesticus). We found that integration of components varied with elaboration of carotenoid-based ornamentation but not of melanin ornamentation. In the house finches, integration was the highest in individuals with small ornaments and decreased with ornament elaboration whereas the pattern was the opposite in common redpolls. These results suggest that in these species integration and complexity of carotenoid-based ornamental components are due to shared condition-dependence of distinct developmental pathways, whereas integration and complexity of the melanin ornamentation is due to organismal integration of developmental pathways and is largely condition- and environment invariant. Thus, functionally, ornamentation of the house sparrows can be considered a single trait, whereas complexity of the house finch and redpoll ornamentation varies with ornament elaboration and individual condition. PMID- 15525417 TI - No place like home: competition, dispersal and complex adaptation. AB - In many groups of organisms the location of settling is determined by competition, and fitter individuals tend to settle closer to their natal territory than less fit ones. In this work we study the implications of this phenomenon to the problem of adaptation and speciation on a rugged adaptive landscape. One consequence of fitness-associated dispersal (FAD) is that individuals with high fitness are more likely to experience inbreeding, especially with other fit individuals. Another consequence is that when dispersal is costly, the less fit individuals are more likely to pay the cost. When a rare and advantageous allelic combination appears, FAD may increase its chances to spread in the population. In a two-locus two-alleles model with negative epistasis, we find that FAD significantly shortens the waiting time for an adaptive peak shift in comparison with random dispersal. PMID- 15525418 TI - In vitro embryo survival and early viability of larvae in relation to male sexual ornaments and parasite resistance in roach, Rutilus rutilus L. AB - According to the 'good genes' hypothesis, sexual ornaments provide an indication of the 'quality' of the bearer. In roach, Rutilus rutilus, breeding tubercles (BTs) may signal resistance against the digenean parasite, Rhipidocotyle campanula. Life history theory predicts that there should be a trade-off between parasite resistance and other life history traits. In roach, this could imply a trade-off between parasite resistance in mature fish and some larval feature. We studied embryo survival and the early viability of larvae of male roach in relation to expression of BTs and parasite resistance in maternal half-sibling families. Highly ornamented males had higher resistance against R. campanula than less ornamented males, but the BTs were not related to either embryo survival or larval viability. However, sires having higher resistance to R. campanula had lower larval viability. These results suggest that BTs of male roach do not indicate 'quality' in terms of early survival or viability, but rather in terms of adult parasite resistance. PMID- 15525419 TI - Divergence in replicated phylogenies: the evolution of partial post-mating prezygotic isolation in bean weevils. AB - By tradition, speciation research has been focused on processes leading to either premating or post-zygotic reproductive isolation. The processes which generate isolation after mating but before zygote formation are less well understood. Here, we study divergence in characters which contribute to post-mating prezygotic isolation, such as egg production and remating rate. We propose that 'replicated' laboratory phylogenies with known histories can be used to yield insights into the processes of divergence. We performed a series of cross-matings between populations within two strains of the bean weevil Callosobruchus maculatus. Each strain has a unique and independent origin and both have been kept in the same set of laboratories during the last few decades. Our results show that divergence has occurred between laboratory populations within strains with regards to the effects that mating has on female reproductive behaviour, showing that the evolution of partial post-mating prezygotic isolation can be rapid. More importantly, the pattern of divergence across populations was distinct in the two strains, suggesting that coevolutionary trajectories are not determined by environmental factors but are to some extent arbitrary. We discuss the limitations of the novel empirical strategy employed here, and conclude that our results lend support to the hypothesis that post-mating sexual selection is capable of rapidly generating post-mating prezygotic isolation. PMID- 15525420 TI - Evolution of sex-biased maternal effects in birds: I. Sex-specific resource allocation among simultaneously growing oocytes. AB - Females in species that produce broods of multiple offspring need to partition resources among simultaneously growing ova, embryos or neonates. In birds, the duration of growth of a single egg exceeds the ovulation interval, and when maternal resources are limited, a temporal overlap among several developing follicles in the ovary might result in a trade-off of resources among them. We studied growth of oocytes in relation to their future ovulation order, sex, and overlap with other oocytes in a population of house finches (Carpodacus mexicanus) where strongly sex-biased maternal effects are favoured by natural selection. We found pronounced differences in growth patterns between oocytes that produced males and females. Male oocytes grew up to five times faster and reached their ovulation size earlier than female oocytes. Early onset and early termination of male oocytes' growth in relation to their ovulation resulted in their lesser temporal overlap with other growing ova compared with female oocytes. Consequently, ovulation mass of female but not male oocytes was strongly negatively affected by temporal overlap with other oocytes. In turn, mass of male oocytes was mostly affected by the order of ovulation and by maternal incubation strategy. These results provide a mechanism for sex-biased allocation of maternal resources during egg formation and provide insights into the timing of the sex determining meiotic division in relation to ovulation in this species. PMID- 15525421 TI - An experimental evaluation of self-interference in Narcissus assoanus: functional and evolutionary implications. AB - Floral traits that reduce self-pollination in hermaphroditic plants have usually been interpreted as mechanisms that limit the genetic consequences of self fertilization. However, the avoidance of sexual conflict between female and male function (self-interference) may also represent an important selection pressure for the evolution of floral traits, particularly in self-incompatible species. Here, we use experimental manipulations to investigate self-interference in Narcissus assoanus, a self-incompatible species with a stigma-height dimorphism in which the degree of spatial separation between sex organs (herkogamy) differs strikingly between the long- and short-styled morphs (hereafter L- and S-morphs). We predicted that weak herkogamy in the L-morph would cause greater self pollination and hence self-interference. Experimental self-pollination reduced seed set when it occurred prior to, or simultaneously with, cross-pollination in the L-morph, but only if it occurred prior to cross-pollination in the S-morph. In the field, autonomous self-pollination was greater in the L-morph than the S morph, but we found no evidence that self-interference reduced maternal or paternal fitness in either morph. One-day-old flowers of the L-morph have reduced stigma receptivity and hence exhibit protandry, whereas stigma receptivity and anther dehiscence are concurrent in the S-morph. This suggests that the two style morphs have alternative strategies for reducing self-interference: dichogamy in the L-morph and herkogamy in the S-morph. These results provide insight into the mechanisms that reduce sexual conflict in hermaphrodite plants and are of significance for understanding the evolution and maintenance of sexual polymorphisms. PMID- 15525422 TI - Testing the status-dependent ESS model: population variation in fighter expression in the mite Sancassania berlesei. AB - The conditional evolutionarily stable strategy (ESS) with status-dependent tactics is the most commonly invoked ESS for alternative reproductive tactics within the sexes. Support for this model has recently been criticized as apparent rather than real. We address key predictions of the status-dependent ESS in three populations of the male dimorphic mite Sancassania berlesei. In S. berlesei'fighter' males are characterized by a thickened pair of legs used for killing rivals; 'scramblers' are benign. Most males in each population could be manipulated to become fighters by decreasing density, fulfilling the prediction that males make a 'decision'. There was evidence of genetic covariance between sire status and offspring morph, but also a strong effect of sire morph on offspring morph ratio. This was consistent with considerable genetic variation for the status-dependent switch point as a breeding experiment found no support for single-locus inheritance. We also found evidence that switch points evolve independently of distributions of status. This study supports the current status dependent ESS model. PMID- 15525423 TI - Predicting evolution of floral traits associated with mating system in a natural plant population. AB - Evolution of floral traits requires that they are heritable, that they affect fitness, and that they are not constrained by genetic correlations. These prerequisites have only rarely been examined in natural populations. For Mimulus guttatus, we found by using the Riska-method that corolla width, anther length, ovary length and number of red dots on the corolla were heritable in a natural population. Seed production (maternal fitness) was directly positively affected by corolla width and anther size, and indirectly so by ovary length and number of red dots on the corolla. The siring success (paternal fitness), as estimated from allozyme data, was directly negatively affected by anther-stigma separation, and indirectly so by the corolla length-width ratio. Genetic correlations, estimated with the Lynch-method, were positive between floral size measures. We predict that larger flowers with larger reproductive organs, which generally favour outcrossing, will evolve in this natural population of M. guttatus. PMID- 15525424 TI - Synergistic epistasis and alternative hypotheses. AB - Inbreeding generally results in deleterious shifts in mean fitness. If the fitness response to increasing inbreeding coefficient is non-linear, this suggests a contribution of epistasis to inbreeding depression. In a cross breeding experiment, Salathe & Ebert (2003. J. Evol. Biol. 16: 976-985) tested and found the presence of this non-linearity in Daphnia magna. They argue that epistatic interactions cause this non-linearity. We argue here that their experimental protocol does not allow disentangling the effect of synergistic epistasis from two alternative hypotheses, namely hybrid vigour and statistical non-independence of data. PMID- 15525426 TI - Introduction: A special issue on cellular biology of protozoan parasites. PMID- 15525427 TI - Contribution of microscopy to a better knowledge of the biology of Giardia lamblia. AB - Giardia lamblia is a flagellated protozoan of great medical and biological importance. It is the causative agent of giardiasis, one of the most prevalent diarrheal disease both in developed and third-world countries. Morphological studies have shown that G. lamblia does not present structures such as peroxisomes, mitochondria, and a well-elaborated Golgi complex. In this review, special emphasis is given to the contribution made by various microscopic techniques to a better knowledge of the biology of the protozoan. The application of video microscopy, immunofluorescence confocal laser scanning microscopy, and several techniques associated with transmission electron microscopy (thin section, enzyme cytochemistry, freeze-fracture, deep-etching, fracture-flip) to the study of the cell surface, peripheral vesicles, endoplasmic reticulum-Golgi complex system, and of the encystation vesicles found in trophozoites and during the process of trophozoite-cyst transformation are discussed. PMID- 15525428 TI - Trichomonads under Microscopy. AB - Trichomonads are flagellate protists, and among them Trichomonas vaginalis and Tritrichomonas foetus are the most studied because they are parasites of the urogenital tract of humans and cattle, respectively. Microscopy provides new insights into the cell biology and morphology of these parasites, and thus allows better understanding of the main aspects of their physiology. Here, we review the ultrastructure of T. foetus and T. vaginalis, stressing the participation of the axostyle in the process of cell division and showing that the pseudocyst may be a new form in the trichomonad cell cycle and not simply a degenerative form. Other organelles, such as the Golgi and hydrogenosomes, are also reviewed. The virus present in trichomonads is discussed. PMID- 15525429 TI - Three-dimensional ultrastructure of the ring stage of Plasmodium falciparum: evidence for export pathways. AB - The three-dimensional structure of the Plasmodium falciparum ring stage has been explored by reconstruction from serial sections and stereoscopic examination of tilted sections. The ring-like light microscopic appearance is related to the shape and contents of the biconcave discoidal parasite at this stage, its thick perimeter containing most of the ribosomes and its thin center containing smooth membrane organelles. The shapes of rings vary between flat and curved cuplike forms. The rough endoplasmic reticulum is a branched network continuous with the nuclear envelope. Evidence for a simple Golgi complex is seen in the presence on the outer nuclear envelope of a locus of coated vesicle budding associated with a single membranous cisterna or cluster of smooth vesicles. In middle and late stage rings this complex migrates along an extension of the nuclear envelope continuous with the rough endoplasmic reticulum. Evidence is also presented for a mechanism of exporting membrane from the parasite into the parasitophorous vacuole membrane and beyond into the red blood cell, by means of double-membraned vesicle-based exocytosis. PMID- 15525430 TI - Polyphosphate content and fine structure of acidocalcisomes of Plasmodium falciparum. AB - Although acidocalcisomes have been well characterized morphologically in other apicomplexan parasites, no such characterization has been done in Plasmodium spp. Here, we report that Plasmodium falciparum merozoites possess electron-dense organelles rich in phosphorus and calcium, as detected by X-ray microanalysis of intact cells, which are similar to the acidocalcisomes of other apicomplexans, but of more irregular form. In agreement with these results malaria parasites possess large amounts of short- and long-chain polyphosphate (polyP), which are associated with acidocalcisomes in other organisms. PolyP levels were highest in the trophozoite stage of the parasite. Treatment of isolated trophozoites with chloroquine resulted in a significant hydrolysis of polyP. Taken together, these results provide evidence that acidocalcisomes from Plasmodium falciparum do not differ significantly from acidocalcisomes of other apicomplexan parasites. PMID- 15525431 TI - Studying the cell biology of apicomplexan parasites using fluorescent proteins. AB - The ability to transfect Apicomplexan parasites has revolutionized the study of this important group of pathogens. The function of specific genes can be explored by disruption of the locus or more subtly by introduction of altered or tagged versions. Using the transgenic reporter gene green fluorescent protein (GFP), cell biological processes can now be studied in living parasites and in real time. We review recent advances made using GFP-based experiments in the understanding of protein trafficking, organelle biogenesis, and cell division in Toxoplasma gondii and Plasmodium falciparum. A technical section provides a collection of basic experimental protocols for fluorescent protein expression in T. gondii. The combination of the in vivo marker GFP with an increasingly diverse genetic toolbox for T. gondii opens many exciting experimental opportunities, and emerging applications of GFP in genetic and pharmacological screens are discussed. PMID- 15525432 TI - Three-dimensional imaging of Toxoplasma gondii-host cell interactions within the parasitophorous vacuole. AB - The protozoan parasite Toxoplasma gondii is a representative of apicomplexan parasites that invades host cells through an unconventional motility mechanism. During host cell invasion it forms a specialized membrane-surrounded compartment that is called the parasitophorous vacuole. The interactions between the host cell and parasite membranes are complex and recent studies have revealed in more detail that both the host cell and the parasite membrane contribute to the formation of the parasitophorous vacuole. By using our a new specimen preparation technique that allows three-dimensional imaging of thick-sectioned internal cell structures with high-resolution, low-voltage field emission scanning electron microscopy, we were able to visualize continuous structural interactions of the host cell membrane with the parasite within the parasitophorous vacuole. Fibrous and tubular material extends from the host cell membrane and is connected to parasite membrane components. Shorter protrusions are also elaborated from the parasite. Several of these shorter fine protrusions connect to the fibrous material of the host cell membrane. The elaborate network may be used for modifications of the parasitophorous vacuole membrane that will allow utilization of nutrients from the host cell by the parasite while it is being protected from host cell attacks. The structural interactions between parasite and host cells undergo time-dependent changes, and a fission pore is the most prominent structure left connecting the parasite with the host cell. The fission pore is anchored in the host cell by thick structural components of unknown nature. The new information gained with this technique includes structural details of fibrous and tubular material that is continuous between the parasite and host cell and can be imaged in three dimensions. We present this technique as a tool to investigate more fully the complex structural interactions of the host cell and the parasite residing in the parasitophorous vacuole. PMID- 15525433 TI - Structural analysis of Cryptosporidium parvum. AB - Cryptosporidium parvum (Apicomplexa, formerly Sporozoa) is the causative agent of cryptosporidiosis, an enteric disease of substantial medical and veterinary importance. C. parvum shows a number of unique features that differ from the rest of the class of coccidea in which it is currently grouped taxonomically. Differences occur in the overall structure of the transmission form and the invasive stages of the parasite, its intracellular location, the presence of recently described additional extracellular stages, the host range and target cell tropism, the ability to autoinfection, the nonresponsiveness to anticoccidial drugs, the immune response of the host, and immunochemical and genetic characteristics. These differences have an important impact on the infectivity, the epidemiology, the therapy, and the taxonomy of the parasite. The present article describes the structural analysis of the parasite using light and electron microscopy with an emphasis on structural details unique to C. parvum. PMID- 15525434 TI - Tissue culture and explant approaches to studying and visualizing Neospora caninum and its interactions with the host cell. AB - Neospora caninum is an apicomplexan parasite first mentioned in 1984 as a causative agent of neuromuscular disease in dogs. It is closely related to Toxoplasma gondii and Hammondia heydorni, and its subsequent description in 1988 has been, and still is, accompanied by discussions on the true phylogenetical status of the genus Neospora. N. caninum exhibits features that clearly distinguish this parasite from other members of the Apicomplexa, including distinct ultrastructural properties, genetic background, antigenic composition, host cell interactions, and the definition of the dog as a final host. Most importantly, N. caninum has a particular significance as a cause of abortion in cattle. In vitro culture has been indispensable for the isolation of this parasite and for investigations on the ultrastructural, cellular, and molecular characteristics of the different stages of N. caninum. Tissue culture systems include maintenance of N. caninum tachyzoites, which represent the rapidly proliferating stage in a large number of mammalian host cells, culture of parasites in organotypic brain slice cultures as a tool to investigate cerebral infection by N. caninum, and the use of techniques to induce the stage conversion from the tachyzoite stage to the slowly proliferating and tissue cyst-forming bradyzoite stage. This review will focus on the use of these tissue culture models as well as light- and electron-microscopical techniques for studies on N. caninum tachyzoites and bradyzoites, and on the physical interactions between parasites and host cells. PMID- 15525435 TI - New approaches to the microscopic imaging of Trypanosoma brucei. AB - Protozoan parasites are fearsome pathogens responsible for a substantial proportion of human mortality, morbidity, and economic hardship. The principal disease agents are members of the orders Apicomplexa (Plasmodium, Toxoplasma, Eimeria) and Kinetoplastida (Trypanosomes, Leishmania). The majority of humans are at risk from infection from one or more of these organisms, with profound effects on the economy, social structure and quality of life in endemic areas; Plasmodium itself accounts for over one million deaths per annum, and an estimated 4 x 10(7) disability-adjusted life years (DALYs), whereas the Kinetoplastida are responsible for over 100,000 deaths per annum and 4 x 10(6) DALYs. Current control strategies are failing due to drug resistance and inadequate implementation of existing public health strategies. Trypanosoma brucei, the African Trypanosome, has emerged as a favored model system for the study of basic cell biology in Kinetoplastida, because of several recent technical advances (transfection, inducible expression systems, and RNA interference), and these advantages, together with genome sequencing efforts are widely anticipated to provide new strategies of therapeutic intervention. Here we describe a suite of methods that have been developed for the microscopic analysis of T. brucei at the light and ultrastructural levels, an essential component of analysis of gene function and hence identification of therapeutic targets. PMID- 15525436 TI - Biogenesis of the reservosomes of Trypanosoma cruzi. AB - Reservosomes are endocytic compartments found in the posterior region of epimastigotes of Trypanosoma cruzi. In the differentiation from trypomastigotes to epimastigotes (reverse metacyclogenesis in vitro), one has the rare opportunity of following the biogenesis of an endocytic compartment. Metacyclic trypomastigotes incubated in LIT medium highly enriched with fetal calf serum differentiated directly to epimastigotes. In recently differentiated epimastigotes, acidic organelles were found in round compartments spread along the cell body, whereas in control epimastigotes they were found in reservosomes located in the posterior region. Ultrastructural analysis of intermediate forms showed that the cytostome and reservosomes appeared before differentiation to epimastigotes was completed. Many polymorphic reservosomes, with or without lipid inclusions, were observed from the anterior portion of the cell body, in close relationship with the Golgi complex, to the posterior region. Endocytic tracers were observed in the cytostome, flagellar pocket, vesicles, and newly formed reservosomes. Cruzipain, the main protease of T. cruzi, was localized in newly formed reservosomes and in vesicles budding from the trans-Golgi network that seem to fuse with reservosomes. Ingested gold-labeled albumin and cruzipain colocalized in recently formed reservosomes. Endocytosis and immunocytochemical analysis suggested that the endocytic and the secretory pathways may contribute to reservosome formation. PMID- 15525437 TI - Acidocalcisomes of Phytomonas francai possess distinct morphological characteristics and contain iron. AB - Acidocalcisomes are acidic calcium storage compartments described initially in trypanosomatid and apicomplexan parasites, and recently found in other unicellular eukaryotes. The aim of this study was to identify the presence of acidocalcisomes in the plant trypanosomatid Phytomonas francai. Electron-dense organelles of P. francai were shown to contain large amounts of oxygen, sodium, magnesium, phosphorus, potassium, calcium, iron, and zinc as determined by X-ray microanalysis, either in situ or when purified using iodixanol gradient centrifugation or by elemental mapping. The presence of iron is not common in other acidocalcisomes. In situ, but not when purified, these organelles showed an elongated shape differing from previously described acidocalcisomes. However, these organelles also possessed a vacuolar H+-pyrophosphatase (V-H+-PPase) as determined by biochemical methods and by immunofluorescence microscopy using antibodies against the enzyme. Together, these results suggest that the electron dense organelles of P. francai are homologous to the acidocalcisomes described in other trypanosomatids, although with distinct morphology and elemental content. PMID- 15525438 TI - Contribution of electron and confocal microscopy in the study of Leishmania macrophage interactions. AB - Promastigotes of the protozoan parasite genus Leishmania are inoculated into a mammalian host when an infected sand fly takes a bloodmeal. Following their opsonization by complement, promastigotes are phagocytosed by macrophages. There, promastigotes differentiate into amastigotes, the form of the parasite that replicates in the phagolysosomal compartments of host macrophages. Although the mechanisms by which promastigotes survive the microbicidal consequence of phagocytosis remain, for the most part, to be elucidated, evidence indicates that glycoconjugates play a role in this process. One such glycoconjugate is lipophosphoglycan, an abundant promastigote surface glycolipid. Using quantitative electron and confocal laser scanning microscopy approaches, evidence was provided that L. donovani promastigotes inhibit phagolysosome biogenesis in a lipophosphoglycan-dependent manner. This inhibition correlates with an accumulation of periphagosomal F-actin, which may potentially form a physical barrier that prevents L. donovani promastigote-containing phagosomes from interacting with endocytic vacuoles. Inhibition of phagosome maturation may constitute a strategy to provide an environment propitious to the promastigote-to amastigote differentiation. PMID- 15525440 TI - Preventing restenosis in early drug-eluting stent era: recent developments and future perspectives. AB - Restenosis is the major limitation of the successful therapy of percutaneous coronary intervention (PCI) for patients with coronary artery disease. The problem was appreciated in the late 1970s to early 1980s. Only in recent years, anti-restenotic therapy has achieved a breakthrough with the development of drug eluting stents. Here, we provide an overview about pathological mechanisms of restenosis after PCI. Present therapeutic approaches to overcome restenosis and recent clinical results are revisited, and some major concerns in the post-drug eluting stent era are discussed. PMID- 15525441 TI - In-vitro transcutaneous delivery of tamoxifen and gamma-linolenic acid from borage oil containing ethanol and 1,8-cineole. AB - The objective of this study was to examine the effects of ethanol and 1,8-cineole on the transcutaneous delivery of tamoxifen and gamma-linolenic acid (GLA) as a two-pronged anti-breast cancer therapy. Formulations containing tamoxifen and varying concentrations of borage oil (approximately 25% GLA), 1,8-cineole and ethanol were prepared and the simultaneous permeation of tamoxifen and GLA determined across full-thickness pig skin using Franz-type diffusion cells over 48 h. Analysis of tamoxifen and GLA (as methyl ester) were by reverse-phase HPLC. The highest flux of tamoxifen of 488.2 +/- 191 x 10(-3) microg cm(-2) h(-1) was observed with a formulation containing 20% 1,8-cineole and 20% ethanol. The same formulation also provided the greatest flux of GLA, 830.6 x 10(-3) microg cm(-2 )h(-1). The findings from this work demonstrate the ability of 1,8-cineole and ethanol to enhance the in-vitro permeation of tamoxifen and GLA across the skin and support the plausibility of simultaneously delivering tamoxifen and GLA transcutaneously as a two-pronged anti-breast cancer system. PMID- 15525442 TI - Evaluation of khaya gum as a directly compressible matrix system for controlled release. AB - Khaya gum has been evaluated as a controlled release agent in modified release matrices in comparison with hydroxypropylmethylcellulose (HPMC) using paracetamol (water soluble) and indometacin (water insoluble) as model drugs. Tablets were produced by direct compression and the in-vitro drug release was assessed in conditions mimicking the gastrointestinal system. Khaya gum matrices provided a controlled release of paracetamol for up to 5 h. The release of paracetamol from khaya gum matrices followed time-independent kinetics (n = 1.042) and release rates were dependent on the concentration of the drug present in the matrix. The addition of tablet excipients not only improved the mechanical properties of the tablet, but also altered the dissolution profile, except for dicalcium phosphate where the profile remained unchanged. HPMC could be used to control the drug release rates from khaya gum matrices and a combination of khaya gum and HPMC gave zero-order time-independent release kinetics. Indometacin exhibited a lag time in excess of 2 h, due to its insolubility at low pH, before the zero-order release was observed. Thus khaya gum matrices could be useful in the formulation of sustained release tablets for up to 5 h and the appropriate combination of khaya gum and HPMC could be used to provide a time-independent release for longer periods. PMID- 15525443 TI - Microsphere-liposome complexes protect adenoviral vectors from neutralising antibody without losses in transfection efficiency, in-vitro. AB - Adenoviral vectors have been commonly used in gene therapy protocols but the success of their use is often limited by the induction of host immunity to the vector. Following exposure to the adenoviral vector, adenoviral-specific neutralising antibodies are produced, which limits further administration. This study examines the effectiveness of a novel combination of microspheres and liposomes for the shielding of adenovirus from neutralising antibodies in an in vitro setting. We show that liposomes are effective in the protection of adenovirus from neutralising antibody and that the conjugation of these complexes to microspheres augments the level of protection. This study further reveals that previously neutralised adenovirus may still be transported into the cell via liposome-cell interactions and is still capable of expressing its genes, making this vector an effective tool for circumvention of the humoral immune response. We also looked at possible side effects of using the complexes, namely increases in cytotoxicity and reductions in transfection efficiency. Our results showed that varying the liposome:adenovirus ratio can reduce the cytotoxicity of the vector as well as increase the transfection efficiency. In addition, in cell lines that are adenoviral competent, transfection efficiencies on par with uncomplexed adenoviral vectors were achievable with the combination vector. PMID- 15525444 TI - Griseofulvin micronization and dissolution rate improvement by supercritical assisted atomization. AB - Supercritical assisted atomization (SAA) was used to micronize griseofulvin (GF), selected as a model compound, to verify the performance of this innovative process. SAA is based on the solubilization of supercritical carbon dioxide in a liquid solution containing the drug. The ternary mixture is then sprayed through a nozzle and microparticles are formed as a consequence of the enhanced atomization. Precipitation temperature and drug concentration in the liquid solution were studied to evaluate their influence on morphology and size of precipitated particles. A good particle size control was obtained and GF spherical particles with mean diameters ranging from 0.5 to 2.5 microm were produced with a narrow particle size distribution. Processed GF was characterized by high-performance liquid chromatography-UV/vis, headspace-gas chromatography flame ionization detection, differential scanning calorimetry, BET and X-ray analyses. No drug degradation was observed and a solvent residue (acetone) less than 800 ppm was measured. GF microparticles showed good stability and surface areas ranging from about 4 to 6 m(2) g(-1); moreover, the micronized drug retained the crystalline habit. GF capsules were formulated with starch and used to compare the dissolution rate of SAA-processed and conventional jet-milled drug. A faster dissolution and a better reproducibility of the dissolution profile were observed for SAA-processed GF. PMID- 15525445 TI - A pharmacokinetic and pharmacodynamic study of desmopressin: evaluating sex differences and the effect of pre-treatment with piroxicam, and further validation of an indirect response model. AB - Desmopressin is a synthetic vasopressin analogue mainly used in treatment of diabetes insipidus and nocturia. Studies in rats have revealed a sex difference in the response to a vasopressin infusion, which was diminished after treatment with an NSAID. This study was performed in man to investigate the influence of sex and concomitant treatment of piroxicam on the pharmacokinetics and dynamics of desmopressin, and to validate a previously described indirect response model. Eight healthy males and eight healthy females participated in the trial, which was conducted in a pharmacokinetic (PK) part followed by a pharmacodynamic (PD) part. Desmopressin was administered intravenously as a single dose (PK = dose 2 microg, PD = dose 0.2 microg). Piroxicam was administered to achieve steady state. The pharmacokinetic parameters of desmopressin were estimated and calculated by means of two-compartmental analysis. In the dynamic part a study design based on an oral hydration model was used. Parameters for urine flow and urine osmolality were estimated. Individual estimates of the pharmacokinetic parameters served as input to the indirect response model that subsequently was fitted to urine osmolality data. The pharmacokinetics of desmopressin after a fixed bolus injection was neither influenced by piroxicam nor sex of the subject. The pharmacodynamics of desmopressin showed a sex difference where females exhibited a more pronounced antidiuretic effect than males, which was statistically significant when the effects were submaximal (>4.5 h after dose). The sex differences were diminished after pre-treatment with piroxicam, indicating a prostaglandin PGE(2)-mediated mechanism. The indirect response model was confirmed, although the modelling could not distinguish a sex difference, indicating a limitation of this model compared with traditional descriptive statistics. PMID- 15525446 TI - Induction of apoptosis in human breast adenocarcinoma MCF-7 cells by prodelphinidin B-2 3,3'-di-O-gallate from Myrica rubra via Fas-mediated pathway. AB - Myrica rubra Sieb et Zucc. (Myricaceae) is well known as a rich source of tannins. Prodelphinidin B-2 3,3'-di-O-gallate (PB233'OG) is a proanthocyanidin gallate that has been reported to exhibit antioxidant and antiviral activity. In this study, we evaluated the anti-proliferative activity of PB233'OG isolated from the bark of M. rubra in human breast adenocarcinoma MCF-7 cells. To identity the anti-cancer mechanism of PB233'OG, we assayed its effect on apoptosis, cell cycle distribution, and levels of p53, p21/WAF1, Fas/APO-1 receptor and Fas ligand. The results showed that PB233'OG induced apoptosis of MCF-7 cells without mediation of p53 and p21/WAF1. We suggest that Fas/Fas ligand apoptotic system is the main pathway of PB233'OG-mediated apoptosis of MCF-7 cells. Our study reports here for the first time that the activity of the Fas/Fas ligand apoptotic system may participate in the anti-proliferative activity of PB233'OG in MCF-7 cells. PMID- 15525447 TI - Role of the carboxylic group in the antispasmodic and vasorelaxant action displayed by kaurenoic acid. AB - The present work describes the investigation of the role of the carboxylic group in the structure-activity relationship of the diterpene ent-kaur-16-en-19-oic acid (kaurenoic acid, KA) in inhibiting rat aorta contraction. For this purpose the methylation of the C-19 carboxyl group of KA was carried out. The effects of the obtained ent-methyl-kaur-16-en-19-oate (KAMe) were compared with those induced by KA. Vascular reactivity experiments showed that KA (50 and 100 microM) concentration-dependently inhibited KCl-induced contraction in both endothelium intact and denuded rat aortic rings. On the other hand, KAMe attenuated KCl induced contraction at 100 microM, but not at 50 microM. KA also reduced CaCl(2) induced contraction in Ca(2+)-free solution containing KCl (30 mM). Again, KAMe produced a less accentuated reduction in CaCl(2)-induced contraction than that induced by the acid KA. KAMe (1-450 microM) concentration-dependently relaxed KCl pre-contracted rings (percentages of relaxation 82.57 +/- 1.65 and 70.55 +/- 4.71, respectively) with denuded endothelium. Similarly, the relaxation induced by KA on phenylephrine (Phe)-pre-contracted rings (73.06 +/- 3.68%) was more pronounced than that found for KAMe (53.68 +/- 4.75%). Pre-incubation of denuded rings for different periods with KA and KAMe showed that the equilibrium periods required by each compound to achieve its maximal inhibitory response on KCl induced contraction are different. Collectively, our results provide functional evidence that methylation of the C-19 carboxyl group of KA reduces but does not abolish the antispasmodic activity displayed by KA. Additionally, we showed that the equilibrium period is a critical step for the inhibitory effect displayed by kaurane-type diterpenes. PMID- 15525448 TI - Red marine alga Bryothamnion triquetrum lectin induces endothelium-dependent relaxation of the rat aorta via release of nitric oxide. AB - We have investigated the vascular relaxant effects of the lectin from a red marine alga Bryothamnion triquetrum (BTL), in particular, the endothelial dependency and the participation of a specific glycoprotein-binding site. BTL (1 100 microg mL(-1)) was applied to rat isolated aortic rings, with or without endothelium, tonically precontracted with phenylephrine (0.1 microM). Endothelium dependent relaxation was assessed in the presence of indometacin (10 microM), L nitro arginine methyl ester (L-NAME, 100 microM) and tetraethylammonium (TEA, 500 microM). For the involvement of the glycoprotein-binding site, BTL was assayed in presence of mucin (300 microg mL(-1)) or N-acetyl D-glucosamine (GlcNAc; 300 microg mL(-1)), a specific and non-specific lectin-binding sugar, respectively. BTL fully and concentration dependently relaxed preparations that possessed an intact endothelium (IC50 (concn producing 50% contraction) = 12.1 +/- 1.6 microg mL(-1)), whereas no significant relaxation was observed in endothelial-denuded tissue. L-NAME, but not indometacin or TEA, completely inhibited the lectin relaxation, suggesting the involvement of nitric oxide (NO). The lectin in association with mucin, but not with GlcNAc, inhibited BTL-induced relaxation, implicating the involvement of the lectin binding site. Our data suggest that the relaxant effect of the red marine alga Bryothamnion triquetrumlectin on isolated aorta occurs via interaction with a specific lectin-binding site on the endothelium, resulting in a release of NO. PMID- 15525449 TI - Investigation into the mast cell stabilizing activity of nature-identical and synthetic indanones. AB - As part of an ongoing search for novel molecules with therapeutic potential we examined the mediator release inhibition activity of a number of indanones and their derivatives. The aldol condensation product 18 was approximately twice as potent as disodium cromoglycate as an inhibitor of compound 48/80-stimulated histamine release from rat peritoneal mast cells. The activity of this class of dimeric indanone compound is significantly higher than controls and may represent a new class of mast cell stabilizing agents. Compound 18 has been selected for further biological evaluation of its mast cell stabilization profile. PMID- 15525450 TI - Carane derivative stereoisomers of different local anaesthetic and antiplatelet activity similarly potentiate forskolin-stimulated cyclic AMP response and bind to beta-adrenoceptors in the rat brain cortex. AB - A carane derivative, KP-23 [RS](-)-4-(2-hydroxy-3)N-isopropylamino)-propoxyimino) cis-carane, was earlier described as a potential local anaesthetic and antiplatelet agent, and the following studies revealed that its R and S stereoisomers, KP-23R and KP-23S, have different potencies in the infiltration anaesthesia and platelet aggregation tests. The effects of these stereoisomers on the cyclic AMP (cAMP) generating system and the displacement of [(3)H]CGP 12177 (a beta-adrenoceptor ligand) from its binding sites in the rat cerebral cortical tissue were investigated. The stereoisomers did not affect the basal cAMP level, but, at concentrations between 10(-4) and 10(-3) M, they elevated the forskolin induced accumulation of cAMP with similar potency. The compounds displaced [(3)H]CGP 12177, however the stereoisomer R was less potent than the racemic KP 23 and the S form (K(i) = 64.1 +/- 5.9 nM, 161.1 +/- 10 nM and 62.1 +/- 5.6 nM for KP-23, KP-23R and KP-23S, respectively). The fact that the stereoisomers differed in both tests only slightly, if at all, suggests that their pharmacological effects are not related to the action on the beta adrenoceptor/adenylate cyclase system. PMID- 15525451 TI - Antihyperglycaemic and antioxidant effect of hyponidd, an ayurvedic herbomineral formulation in streptozotocin-induced diabetic rats. AB - Hyponidd is a herbomineral formulation composed of the extracts of ten medicinal plants ( Momordica charantia, Melia azadirachta, Pterocarpus marsupium, Tinospora cordifolia , Gymnema sylvestre, Enicostemma littorale, Emblica officinalis, Eugenia jambolana, Cassia auriculata and Curcuma longa). We have investigated hyponidd for its possible antihyperglycaemic and antioxidant effect in diabetic rats. Rats were rendered diabetic by streptozotocin (STZ) (45 mg kg(-1) body weight). Oral administration of hyponidd (100 mg kg(-1) and 200 mg kg(-1)) for 45 days resulted in significant lowered levels of blood glucose and significant increased levels of hepatic glycogen and total haemoglobin. An oral glucose tolerance test was also performed in experimental diabetic rats in which there was a significant improvement in blood glucose tolerance in the rats treated with hyponidd. Hyponidd administration also decreased levels of glycosylated haemoglobin, plasma thiobarbituric acid reactive substances, hydroperoxides, ceruloplasmin and alpha-tocopherol in diabetic rats. Plasma reduced glutathione and vitamin C were significantly elevated by oral administration of hyponidd. The effect of hyponidd at a dose of 200 mg kg(-1) was more effective than glibenclamide (600 microg kg(-1)) in restoring the values to near normal. The results showed that hyponidd exhibits antihyperglycaemic and antioxidant activity in STZ-induced diabetic rats. PMID- 15525452 TI - Corni fructus as the major herb of Die-Huang-Wan for lowering plasma glucose in Wistar rats. AB - Die-Huang-Wan is a mixture of six herbs used to lower plasma glucose by increasing insulin secretion in normal rats. Die-Huang-Wan contains the herbs dioscorea (Dioscoreae rhizoma), cornus (Corni fructus), alisma (Rhizoma alismatis), holelen (Poria), rehmannia (Rehmanniae radix) and tree peony bark (Moutan radicis cortex). The present study was designed to clarify the major herb contributing to the plasma glucose-lowering action of Die-Huang-Wan in rats. A decrease in plasma glucose was not observed in Wistar rats treated with the cornus-deleted formula of Die-Huang-Wan; however, the action was retained in the other herb-deleted formulas containing cornus. In normal rats, the decrease in plasma glucose and increase in plasma insulin concentrations were dependent on the dose of cornus and were similar to those produced by Die-Huang-Wan. Treatment of Wistar rats with each of the other five herbs separately did not result in a decrease in plasma glucose. Moreover, the increase in plasma insulin or reduction in plasma glucose resulting from cornus treatment was blocked by atropine or 4 diphenylacetoxy-N-methylpiperidine methiodide mustard, indicating mediation of muscarinic M(3) receptors similar to that caused by Die-Huang-Wan. These results suggest that cornus is the major contributor to the plasma glucose-lowering action in Die-Huang-Wan in normal rats. PMID- 15525453 TI - Hepatoprotective activity of Terminalia catappa L. leaves and its two triterpenoids. AB - The aim of this study was to evaluate the effect of the chloroform extract of Terminalia catappa L. leaves (TCCE) on carbon tetrachloride (CCl(4))-induced acute liver damage and D-galactosamine (D-GalN)-induced hepatocyte injury. Moreover, the effects of ursolic acid and asiatic acid, two isolated components of TCCE, on mitochondria and free radicals were investigated to determine the mechanism underlying the action of TCCE on hepatotoxicity. In the acute hepatic damage test, remarkable rises in the activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) (5.7- and 2.0-fold) induced by CCl(4) were reversed and significant morphological changes were lessened with pre treatment with 50 and 100 mg kg(-1) TCCE. In the hepatocyte injury experiment, the increases in ALT and AST levels (1.9- and 2.1-fold) in the medium of primary cultured hepatocytes induced by D-GalN were blocked by pre-treatment with 0.05, 0.1, 0.5 g L(-1) TCCE. In addition, Ca(2+)-induced mitochondrial swelling was dose-dependently inhibited by 50-500 microM ursolic acid and asiatic acid. Both ursolic acid and asiatic acid, at concentrations ranging from 50 to 500 microM, showed dose-dependent superoxide anion and hydroxyl radical scavenging activity. It can be concluded that TCCE has hepatoprotective activity and the mechanism is related to protection of liver mitochondria and the scavenging action on free radicals. PMID- 15525454 TI - In-vitro vasodilatory activity of the hexanic extract of leaves and stems from Piper truncatum Vell. in rats. AB - Several plants from the Brazilian Tropical Forest are used in folk medicine for treatment of hypertension and asthma. In this study, we investigated the effects of hexanic extracts of leaves (HLE) and stems (HSE) from Piper truncatum on the contractility of cardiac, vascular and tracheal smooth muscles. Twitches of cardiac muscles obtained with electrical stimulation were recorded before and after exposure to increasing concentrations of hexanic extracts. HLE and HSE respectively reduced significantly the amplitude of twitches to 57.05 +/- 11.63 and 61.58 +/- 5.70% of control in the presence of 100 microg mL(-1). Contractile response to a single concentration of adrenaline (epinephrine) was measured before and after exposure of rat aorta rings to HLE and HSE. Both extracts inhibited aorta contraction in a concentration-dependent manner. The concentration of 50% inhibitory effect (IC50) was 32.3 +/- 13.8 and 47.0 +/- 23.8 microg mL(-1) for HLE and HSE, respectively, in aorta with intact endothelium. HLE and HSE also reduced the acetylcholine-precontracted trachea in a concentration-dependent manner with maximal effect observed at 250 and 350 microg mL(-1), respectively. Vasodilatation and trachea relaxation induced by HLE and HSE could explain the use of Piper extracts to reduce blood pressure and bronchospasm. PMID- 15525455 TI - The combination of modified antioxidant enzymes for anti-thrombotic protection of the vascular wall: the significance of covalent connection of superoxide dismutase and catalase activities. AB - Vascular wall protection can be achieved by preventive attachment to the vascular wall of antioxidants and elimination/neutralization of toxic products after their disproportioning. For this purpose we have prepared covalent conjugates between the vascular wall glycosaminglycan chondroitin sulfate (CHS) and the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The following conjugates were obtained: SOD-CHS, CAT-CHS and SOD-CHS-CAT. Their anti-thrombotic activity was compared in a rat model of arterial thrombosis by measuring the time of occlusion emergence and thrombus mass. It is noteworthy that the effectiveness of single bolus injections of SOD-CHS/CAT-CHS mixture was much lower than that of the bienzymic SOD-CHS-CAT conjugate. The conjugate SOD-CHS-CAT proved to be anti thrombotically effective in doses two orders of magnitude lower than the native biocatalysts and an order of magnitude lower than SOD-CHS and CAT-CHS derivatives. For effective anti-thrombotic protection in oxidative conditions it is important to maintain the stable connection of SOD and CAT activity on the vascular wall and the large size of these conjugates. Covalent conjugate SOD-CHS CAT is the best prospect for pharmaceutical development. PMID- 15525456 TI - Metabolism of the dihydropyridine calcium channel blockers mebudipine and dibudipine by isolated rat hepatocytes. AB - The prototype 1,4-dihydropyridine (1,4-DHP) nifedipine, indicated for the management of hypertension and angina pectoris, has drawbacks of rapid onset of vasodilating action and a short half-life. Several newer analogues have been designed to offset these problems and these include mebudipine and dibudipine. These analogues contain t-butyl substituents that have been selected to alter the fast metabolism without altering pharmacological activity. In this study, the metabolism of mebudipine and dibudipine by isolated rat hepatocytes has been investigated. These compounds were extensively metabolized in 2 h by oxidative pathways, analogous to those known for nifedipine, and by O-glucuronidation after hydroxylation of the t-butyl substituents. The in-vitro half-lives of mebudipine (22 +/- 7.1 min) and dibudipine (40 +/- 9.8 min) were significantly longer than that of nifedipine (5.5 +/- 1.1 min), which was investigated in parallel in this study. These newer 1,4-DHPs address the problem of the short half-life of nifedipine and have potential for further development in view of their comparable potency to nifedipine. PMID- 15525457 TI - Anti-tumor and immunoregulatory activities of Ganoderma lucidum and its possible mechanisms. AB - Ganoderma lucidum (G lucidum) is a medicinal fungus with a variety of biological activities. It has long been used as a folk remedy for promotion of health and longevity in China and other oriental countries. The most attractive character of this kind of medicinal fungus is its immunomodulatory and anti-tumor activities. Large numbers of studies have shown that G lucidum modulate many components of the immune system such as the antigen-presenting cells, NK cells, T and B lymphocytes. The water extract and the polysaccharides fraction of G lucidum exhibited significant anti-tumor effect in several tumor-bearing animals mainly through its immunoenhancing activity. Recent studies also showed that the alcohol extract or the triterpene fraction of G lucidum possessed anti-tumor effect, which seemed to be related to the cytotoxic activity against tumor cells directly. Preliminary study indicated that antiangiogenic effect may be involved antitumor activity of G lucidum. PMID- 15525458 TI - Tissue distribution of bitespiramycin and spiramycin in rats. AB - AIM: To investigate the tissue distribution of bitespiramycin (BSPM) and spiramycin (SPM) in rats. METHODS: Liquid chromatographic-mass spectrometric assay was applied for the determination of three major components (isovalerylspiramycins, ISV-SPMs) of BSPM and their major active metabolites (SPMs) in rat tissues and plasma after an oral dose of bitespiramycin, as well as SPMs. RESULTS: High levels of drug concentrations were observed in most tissues, especially in the liver, stomach, intestine, spleen, lung, womb, and pancreas. BSPM persisted long time in many rat tissues such that the drug concentration in spleen was 69.4 nmol/g at 60 h post-dose and it was still above the minimum inhibitory concentration of many susceptible pathogens. At 2.5 h post-dose, the total concentrations of ISV-SPMs and SPMs achieved in tissues were from 6 to 215 times higher than the corresponding concentrations in plasma. At 2.5 h post-dose, the mean C(t)/C(p) of BSPM appeared to be 2- or 3-fold those of SPM in most tissues. The tissue to plasma concentration ratios following oral dose of BSPM were higher than those of SPM in most tissues. The drug was not detected in brain and testis after a single dose of BSPM and SPM. CONCLUSION: Both BSPM and SPM penetrate into rat tissues well and BSPM has higher tissue affinity than SPM. PMID- 15525459 TI - Pharmacokinetics of magnesium lithospermate B after intravenous administration in beagle dogs. AB - AIM: To develop a specific liquid chromatography-tandem mass spectrometry (LC MS/MS) method for the pharmacokinetic study of magnesium lithospermate B (MLB), and study the pharmacokinetics of MLB after i.v. administration in beagle dogs. METHODS: Each beagle dog was iv administered MLB 3, 6, and 12 mg/kg random. The serum drug concentration was determined by specific liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays. The pharmacokinetic parameters were calculated by Drug and Statistics version 1.0 program. RESULTS: The calibration curve for MLB was linear over a range of 16-4096 microg/L with coefficients of correlation >0.999. The intra- and inter-day precisions (CV) of analysis were <10 %, and accuracy ranged from 90 % to 113 %. After i.v. administration of MLB at the doses of 3, 6, and 12 mg/kg, the C(0) values for MLB were estimated to be of 24, 47, and 107 mg/L, respectively. The AUC increased with the increasing doses for iv administration, and the mean AUC(0-t) values were 109.3, 247.9, and 582.4 mg x min x L(-1), respectively. MLB was distributed and eliminated quickly from central compartment, the mean T(1/2alpha) values for MLB at doses of 3, 6, 12 mg/kg were 2.2, 2.7, and 2.9 min, and the mean T(1/2beta) values were 43, 42, and 42 min, respectively. CONCLUSION: This LC-MS/MS method is rapid, sensitive, and specific for the pharmacokinetic study of MLB. The kinetic process of MLB in beagle dogs in vivo was best fitted to a two-compartment model. For i.v. administration, the pharmacokinetic parameters of C(0) and AUC have good linearity among the doses, and MLB was distributed and eliminated quickly in beagle dogs. PMID- 15525460 TI - Moclobemide up-regulates proliferation of hippocampal progenitor cells in chronically stressed mice. AB - AIM: To explore the action mechanism of antidepressants. METHODS: The PC12 cell proliferation was detected by flow cytometry. The proliferation of hippocampal progenitor cells and level of brain-derived neurotrophic factor (BDNF) were measured by immunohistochemistry. RESULTS: Treatment with N-methylaspartate (NMDA) 600 micromol/L for 3 d significantly decreased the percentage of S-phase in PC12 cells, while in the presence of classical antidepressant, moclobemide (MOC) 2 and 10 micromol/L, the percentage in S-phase increased. Furthermore, the proliferation of progenitor cells in hippocampal dentate gyrus (subgranular zone), as well as the level of BDNF in hippocampus significantly decreased in chronically stressed mice, while chronic administration with MOC 40 mg/kg (ip) up regulated the progenitor cell proliferation and BDNF level in the same time course. CONCLUSION: Up-regulation of the proliferation of hippocampal progenitor cells is one of the action mechanisms for MOC, which may be closely related to the elevation of BDNF level at the same time. These results also extend evidence for our hypothesis that up-regulation of the hippocampal neurogenesis is one of the common mechanisms for antidepressants. PMID- 15525461 TI - A new buprenorphine analogy, thenorphine, inhibits morphine-induced behavioral sensitization in mice. AB - AIM: To investigate effects of thenorphine, a new compound of partial agonist of mu-opioid receptor, on the locomotor activity and the behavioral sensitization to morphine in mice. METHODS: Locomotor activity was observed after administration of thenorphine or co-administration of thenorphine and morphine in mice. Mice were induced behavioral sensitization to morphine by intraperitoneal injection of 20 mg/kg morphine once daily for 7 d. Thenorphine was co-administrated with morphine to observe the effects of thenorphine on the development, transfer and expression of morphine-induced behavioral sensitization. RESULTS: A single dose of thenorphine (0.0625, 0.25, and 1.0 mg/kg) could dose-dependently inhibit the locomotor activity in mice (P<0.05), repeated administrations of thenorphine, however, were not able to induce locomotor sensitization, but induced tolerance. Pretreatment with thenorphine 30 min prior to morphine effectively inhibited the psychomotor effect of morphine in mice (P<0.01). Co-administration of thenorphine (0.0625, 0.25, and 1.0 mg/kg) could dose-dependently inhibit the development, transfer, and expression of behavioral sensitization to morphine in mice (P<0.05 or P<0.01). CONCLUSION: Thenorphine inhibited morphine-induced behavioral sensitization in mice, suggesting that thenorphine may be effective against the addiction of opioids. PMID- 15525462 TI - Homocysteine induces production of monocyte chemoattractant protein-1 and interleukin-8 in cultured human whole blood. AB - AIM: To investigate whether increased plasma L-homocysteine (Hcy) level could promote monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in cultured whole blood. METHODS: Human whole blood or different type of peripheral blood cells from health volunteers were incubated with Hcy and/or the inhibitors. MCP-1 and IL-8 level were measured by ELISA assay. RESULTS: Hcy 10-1000 micromol/L induced production of MCP-1 and IL-8 in cultured human whole blood (P<0.05). The major cellular source of these chemokines comed from monocytes. Meanwhile,Hcy also promoted the upregulation of MPO level even at the 10 micromol/L in the cultured whole blood. The intracellular ROS, particular the OH. radicals, play extremely important role in the Hcy-induced MCP-1 and IL-8 production. CONCLUSION: Increased Hcy level in plasma (hyperhomocysteinemia) induced MCP-1 and IL-8 secretion in cultured human whole blood, especially in monocytes via oxidative stress mechanism. PMID- 15525463 TI - Losartan inhibited expression of matrix metalloproteinases in rat atherosclerotic lesions and angiotensin II-stimulated macrophages. AB - AIM: To explore whether the angiotensin II (Ang II) receptor 1 (AT1) antagonist, losartan could reduce activity and expression of matrix metalloproteinases (MMPs) in rat atherosclerotic plaques. METHODS: Male Wistar-Kyoto rats were ip injected a single dose of vitamin D3 600 kU x kg(-1) x month(-1) and fed an atherogenic diet for 4 months to induce experimental atheroma. Then either placebo or losartan 50 mg x kg(-1) x d(-1) was administered in rats for another 2 months. In vitro, the effect of losartan 0.1-10 micromol/L on the expression of MMP-2 and MMP-9 was investigated in Ang II-stimulated rat peritoneal macrophages. The expression and activity of MMP-2 and MMP-9 were monitored by Western blot, RT PCR, and SDS-PAGE zymography analysis. RESULTS: High levels of MMP-2 and MMP-9 were expressed in rat atherosclerotic lesions. Losartan significantly reduced the activity and expression of MMP-2 and MMP-9 compared with the placebo group (MMP 2, 5861+/-539 vs 8991+/-965, P<0.05; MMP-9,10527+/-1002 vs 14623+/-2462, P<0.01). In cultured rat peritoneal macrophages, Ang II 0.1 micromol/L elicited an increase in MMP-2 and MMP-9 activity and expression that were prevented by losartan in a dose-dependent manner (P<0.01). But the AT2 receptor antagonist PD123319 had no effect. CONCLUSION: Losartan reduced the expression and activity of MMP-2 and MMP-9 in rat atherosclerotic lesions. The anti-atherogenic effects of losartan were due to the direct inhibition of Ang II bioactivity. PMID- 15525464 TI - AT1 receptor in rostral ventrolateral medulla mediating blunted baroreceptor reflex in spontaneously hypertensive rats. AB - AIM: To determine the role of AT1 receptor in the rostral ventrolateral medulla (RVLM) in mediating the blunted baroreceptor reflex in spontaneously hypertensive rats (SHR). METHODS: Intravenous injections of graded doses of phenylephrine (1, 5, 10, 20, and 40 microg/kg) increased the blood pressure to elicit the baroreceptor reflex in both SHR and normotensive Wistar rats anesthetized with urethane. The baroreceptor reflex sensitivity (BRS) was determined before and after microinjection of Ang II, losartan, or AT1 receptor antisense oligodeoxynucleotides into the RVLM. AT1 receptor protein level in the RVLM was measured by Western blotting. RESULTS: The BRS was significantly decreased in SHR compared with normal rats. Bilateral microinjection of AT1 receptor antagonist losartan (250 nmol/h) into the RVLM partly reversed the blunted BRS in SHR, but had no significant effect on the BRS in normal rats. Ang II (1.5 nmol/h) significantly inhibited the BRS in normal rats, which was completely abolished by pretreatment with losartan. However, no significant change in the BRS was observed after microinjection of Ang II into the RVLM in SHR. Bilateral microinjection of AT1 receptor antisense oligodeoxynucleotides (ASODN) into the RVLM partially recovered the blunted BRS in SHR after 3 h, but no significant change in the BRS was observed in normal rats. The AT1 receptor protein level significantly decreased after administration of ASODN. CONCLUSION: Blockage of AT1 receptor or inhibition of AT1 receptor protein synthesis in the RVLM enhanced the BRS in SHR, suggesting that the enhanced activities of AT1 receptor in the RVLM contribute to the blunted BRS in SHR. PMID- 15525465 TI - Capsaicin facilitates carotid sinus baroreceptor activity in anesthetized rats. AB - AIM: To study the effect of capsaicin on carotid sinus baroreceptor activity (CBA). METHODS: The functional curve of carotid baroreceptor (FCCB) was constructed and the functional parameters of carotid sinus baroreceptor were measured by recording sinus nerve afferent discharge in anesthetized rats with perfused isolated carotid sinus. RESULTS: Low-concentration of capsaicin (0.2 mumol/L) had no significant effect on CBA, while perfusion of the isolated carotid sinus with middle-concentration of capsaicin (1 mumol/L) could shift FCCB to the left and upward, with peak slope (PS) increased from (2.47 %+/-0.14 %)/mmHg to (2.88 %+/-0.10 %)/mmHg (P<0.05) and peak integral value of carotid sinus nerve discharge (PIV) enhanced from 211 %+/-5 % to 238 %+/-6 % (P<0.01). The threshold pressure (TP) and saturation pressure (SP) were significantly decreased from 68.0+/-1.1 to 62.7+/-1.0 mmHg (P<0.01) and from 171.0+/-1.6 to 165.0+/-0.6 mmHg (P<0.01). By perfusing with high-concentration of capsaicin (5 micromol/L), FCCB was shifted to the left and upward further and the changes of the functional parameters such as PS, TP, and SP were concentration-dependent. Pretreatment with ruthenium red (100 micromol/L), an antagonist of vanilloid receptor subtype 1 (VR1), blocked the effect of capsaicin on CBA. Preperfusion with glibenclamide (20 micromol/L), a K(ATP) channel blocker, could eliminate the effect of capsaicin on CBA. CONCLUSION: Capsaicin exerts a facilitatory role on the isolated carotid baroreceptor in a concentration-dependent manner. The facilitatory action of capsaicin may be attributed to the opening of K(ATP) channels mediated by VR1. PMID- 15525466 TI - Effects of rat urotensin II on coronary flow and myocardial eNOS protein expression in isolated rat heart. AB - AIM: To examine the effects of urotensin II, a recently discovered endogenous peptide, on coronary flow (CF), cardiac function, and endothelial nitric oxide synthase (eNOS) expression in isolated rat hearts. METHODS: Heart was isolated and perfused retrogradely via the aorta in Langendorff mode. Rat urotensin II was administered in the perfusion solution. The eNOS content in myocardium was determined by Western blot. RESULTS: Rat urotensin II had no effect on the heart rate, left ventricular systolic pressure, left ventricular end-diastolic pressure, or +/-dp/dt(max). While rat urotensin II dose-dependently increased CF. CF was increased by 11.43 %, 6.67 %, 6.62 %, 6.56 %, 6.36 %, and 5.86 % respectively in a time-dependent manner at 5, 10, 15, 20, 25, and 30 min after injection of rat urotensin II 6.66 x 10(-2) microg. The maximal effect on CF was found at 5 min following urotensin II administration. N(G)-nitro-L-arginine methyl ester (L-NAME) did not prevent the increased CF in response to urotensin II. Rat urotensin II dose-dependently increased the cardiac eNOS protein expression and this effect was not inhibited by L-NAME. CONCLUSION: Rat urotensin II did not alter cardiac function but increased CF and the amount of myocardial eNOS protein in the isolated rat heart. The increased CF was independent of the involvement of eNOS. PMID- 15525467 TI - Muscarinic cholinergic regulation of L-type calcium channel in heart of embryonic mice at different developmental stages. AB - AIM: To investigate the muscarinic regulation of L-type calcium current (I(Ca-L)) during development. METHODS: The whole cell patch-clamp technique was used to record II(Ca-L) in mice embryonic cardiomyocytes at different stages (the early developmental stage, EDS; the intermediate developmental stage, IDS; and the late developmental stage, LDS). Carbachol (CCh) was used to stimulate M-receptor in the embryonic cardiomyocytes of mice. RESULTS: The expression of I(Ca-L) density did not change in different developmental stages (P>0.05). There was no difference in the sensitivity of I(Ca-L) to CCh during development (P>0.05). This inhibitory action of CCh was mediated by inhibition of cyclic AMP since 8-bromo cAMP completely reversed the muscarinic inhibitory action. IBMX, a non-selective inhibitor of phosphodiesterase (PDE), reversed the inhibitory action of M receptor on I(Ca-L) current by 71.2 %+/-9.2 % (n=8) and 11.3 %+/-2.5 % (n=9) in EDS and LDS respectively. However forskolin, an agonist of adenylyl cyclase (AC), reversed the action of CCh by 14.5 %+/-3.5 % (n=5) and 82.7 %+/-10.4 % (n=7) in EDS and LDS respectively. CONCLUSION: The inhibitory action of CCh on I(Ca-L) current was mediated in different pathways: in EDS, the inhibitory action of M receptor on I(Ca-L) channel mainly depended on the stimulation of PDE. However, in LDS, the regulation by M-receptor on I(Ca-L) channel mainly depended on the inactivation of AC. PMID- 15525468 TI - Effects of imidapril on heterogeneity of action potential and calcium current of ventricular myocytes in infarcted rabbits. AB - AIM: To investigate the effects of chronic treatment with imidapril on the electrophysiologic heterogeneous change of the noninfarcted myocardium of rabbits after myocardial infarction and the mechanism of its antiarrhythmic efficacy. METHODS: Rabbits with left coronary artery ligation were prepared and allowed to recover for 8 weeks. Myocytes were isolated from subendocardial, midmyocardial, and subepicardial regions of the noninfarcted left ventricular wall. Action potentials and calcium current were recorded using whole-cell patch clamp technique. RESULTS: The action potential duration of repolarization 90 % (APD90) was more prolonged in midmyocardium rather than in subepicardium and subendocardium with healed myocardial infarction. The transmural dispersion of repolarization (TDR) was increased in the three ventricular regions. The amplitude of I(Ca-L) [was enhanced but its density was decreased in noninfarcted ventricular myocytes due to increased cell membrane capacitance. The increased differences of calcium currents among subepicardium, midmyocardium, and subendocardium were also discovered. Normalization of heterogeneous changes in repolarization after treatment with imidapril was observed and decrease of TDR in noninfarcted area was measured. Early after depolarization (EAD) events of noninfarcted midmyocardium were markedly decreased by imidapril. CONCLUSION: Imidapril reduced the electrophysiologic heterogeneities in noninfarcted area in rabbits after myocardial infarction. This ability of imidapril may contribute to its antiarrhythmic efficacy. PMID- 15525469 TI - Construction and activity of a novel GHRH analog, Pro-Pro-hGHRH(1-44)-Gly-Gly Cys. AB - AIM: To construct another growth hormone releasing hormone (GHRH) analog, Pro-Pro hGHRH(1-44)-Gly-Gly-Cys peptide and to compare its activity with that of Pro-Pro hGHRH(1-44)OH peptide. METHODS: The pro-pro-hGHRH(1-44)-gly-gly-cys DNA fragment was synthesized by polymerase chain reaction. The recombinant protein was expressed to high levels in Escherichia coli BL21 (DE3). The Pro-Pro-hGHRH (1-44) Gly-Gly-Cys peptide was purified to homogeneity by cell disruption, washing, ethanol precipitation, acid hydrolysis, and SP-Sephadex C-25 and Sephadex G-25 column chromatography. The peptide molecular mass was determined by electrospray ionization mass spectroscopy (ESI-MS). Its purity was determined by SDS-PAGE. The concentration of growth hormone (GH) stimulated by GHRH and its analogs was determined with antiserum kit against human GH. The other human hormones as hTSH, hFSH, hLH, and hPRL were determined with a paramagnetic particle chemiluminescent immunoassay kit. RESULTS: The molecular weight of Pro-Pro-hGHRH (1-44)-Gly-Gly Cys was 5455.4 kDa which was coincident with the theoretical calculations. All the three peptides at 5 mg/L stimulated GH release from the human fetal pituitary but only the difference between Pro-Pro-hGHRH (1-44)-Gly-Gly-Cys group and blank group was significant (P<0.05). Pro-Pro-hGHRH (1-44)-Gly-Gly-Cys 0.01, 0.1, and 1 mg/L and Pro-Pro-hGHRH (1-44)OH 0.1 and 1 mg/L stimulated GH release from rat pituitary in a concentration-dependent manner (P<0.05, P<0.01). At the same concentration Pro-Pro-hGHRH (1-44)-Gly-Gly-Cys stimulated more GH release than Pro-Pro-hGHRH (1-44)OH. Pro-Pro-hGHRH (1-44)OH 0.01 mg/L and hGHRH (1-40)OH 2 mg/L did not stimulate GH release from rat pituitary (P>0.05). Pro-Pro-hGHRH (1 44)OH and Pro-Pro-hGHRH (1-44)-Gly-Gly-Cys peptides at 5 mg/L did not stimulate hTSH, hFSH, hLH, and hPRL release from human fetal pituitary in vitro. CONCLUSION: Pro-Pro-hGHRH (1-44)-Gly-Gly-Cys had a better activity than that of Pro-Pro-hGHRH(1-44)OH. Variation at C-terminus of GHRH could modulate its GH releasing activity. PMID- 15525470 TI - Yeast two-hybrid screening for proteins that interact with alpha1-adrenergic receptors. AB - AIM: To find novel proteins that may bind to alpha1A-adrenergic receptor (alpha1A AR) and investigate their interactions with the other two alpha1-AR subtypes (alpha1B-AR and alpha1D-AR) with an expectation to provide new leads for the function study of the receptors. METHODS: Yeast two-hybrid assay was performed to screen a human brain cDNA library using the C terminus of alpha1A-AR (alpha1A-AR CT) as bait. X-Gal assay and o-nitrophenyl-beta-D-galactopyranoside (ONPG) assay were subsequently conducted to further qualitatively or quantitatively confirm the interactions between receptors and the three identified proteins. RESULTS: (1) Selection medium screening identified segments of bone morphogenetic protein 1 (BMP-1), active Bcr-related protein (Abr), and filamin-C as binding partners of alpha1A-AR-CT in yeast cells respectively. Besides, protein segments of BMP-1 and Abr could only specifically interact with alpha1A-AR-CT while filamin-C segment interacted with all three alpha1-AR subtypes. (2) In X-Gal assay, the co transformants of alpha1A-AR-CT and BMP-1 segments turned strong blue at about 30 min while other positive transformants only developed weak blue at about 5-6 h. (3) In ONPG assay, interaction (shown in beta-galactosidase activity) between alpha1A-AR-CT and BMP-1 segments was about 30 times stronger than that of control (P<0.01), while other positive interactions were only about 2-5 times as strong as those of controls (P<0.05). CONCLUSION: In yeast cells BMP-1, Abr and/or filamin-C could interact with three alpha1-AR subtypes, among which, interaction between BMP-1 and alpha1A-AR was the strongest while other interactions between proteins and receptors were relatively weak. PMID- 15525471 TI - Effect of IFN-gamma and dexamethasone on TGF-beta1-induced human fetal lung fibroblast-myofibroblast differentiation. AB - AIM: To study whether Smads signaling pathway was involved in human fetal lung fibroblast-myofibroblast differentiation induced by transforming growth factor (TGF)-beta1 and the role of interferon (IFN)-gamma, dexamethasone (DEX) in the fibroblast-myofibroblast differentiation. METHODS: Alpha-smooth muscle actin (alpha-SMA), Smad2/3, and Smad7 protein were assessed by Western blot. Collagen protein was analyzed by measuring hydroxyproline. Alpha-SMA and collagen III mRNA were assessed by RT-PCR. Myofibroblasts morphology and Smad2/3 nuclear translocation were assessed by immunohistochemistry. The overexpression of Smad7, a negative mediator of Smads signaling pathway, was acquired by transfection of Smad7 vector. RESULTS: During fibroblast-myofibroblast differentiation induced by TGF-beta1, IFN-gamma 200 microg/L markedly blocked TGF-beta1-induced alpha-SMA protein expression (P<0.01), collagen protein (P<0.01) and mRNA (P<0.05) expression, and myofibroblasts morphological transformation, but DEX 10 micromol/L augmented TGF-beta1-induced alpha-SMA expression (P<0.01). For myofibroblasts, both IFN-gamma 200 microg/L and DEX 10 micromol/L did not inhibit alpha-SMA expression (P>0.05) and collagen protein (P>0.05) and mRNA expression (P>0.05) and did not change myofibroblasts morphology. Transient transfection of Smad7 vector resulted in significant inhibition of TGF-beta1-induced alpha-SMA expression (P<0.01). IFN-gamma 200 microg/L did not block TGF-beta1-stimulated Smad2/3 phosphorylation and their nuclear translocation. CONCLUSION: TGF-beta1 induced fibroblast-myofibroblast differentiation in a Smad proteins-dependent manner. IFN-gamma could block this process but it was not mediated by interrupting smad2/3 phosphorylation and their nuclear translocation and DEX played a synergism with TGF-beta1. Differentiated myofibroblasts, however, were resistant to both IFN-gamma and DEX. PMID- 15525472 TI - Effects of estrogen on gastrocnemius muscle strain injury and regeneration in female rats. AB - AIM: To study the effects of estrogen on muscle damage and regeneration after acute passive gastrocnemius muscle strain injury in female Sprague-Dawley rats. METHODS: Rats were divided into 5 groups: ovariectomized, strained and treated with low-dosage estradiol (20 microg/d) (E(low)), treated with high-dosage estradiol (200 microg/d) (E(high)), treated with oil placebo (Oil), strained with no ovariectomy (Strain), and sham operated with no strain and no ovariectomy (Con). Muscle damage index [plasma creatine kinase (CK)], antioxidant indexes [glutathione (GSH), Vitamin E (Vit E), total antioxidant capability (TAC)], and muscle regeneration index (desmin) were investigated at 7 d. RESULTS: The plasma CK activity increased but GSH, Vit E, and TAC levels decreased after muscle strain injury (Strain vs Con P<0.05). Plasma CK activity was the greatest while GSH, Vit E, and TAC were the lowest in the Oil group among the five groups (P<0.01). Plasma CK in the E(high) and Strain groups was lower than that in the E(low) group. Plasma GSH, Vit E, and TAC were higher in the E(high) and Strain groups compared with the E(low) group (P<0.05). The expression of desmin in the E(high) and Strain groups was higher than that in the E(low) group (P<0.01) while that in the Oil group was the lowest in all the five groups (P<0.01). CONCLUSION: Endogenous estrogen in normal female rats or exogenous estrogen in ovariectomized rats could improve antioxidant capability in vivo, so that reduced muscle damage and accelerated muscle regeneration post gastronemius muscle strain injury. PMID- 15525473 TI - Therapeutic effects of glucosides of Cheanomeles speciosa on collagen-induced arthritis in mice. AB - AIM: To investigate the therapeutic effect of the glucosides of Cheanomeles speciosa (GCS) on the collagen-induced arthritis (CIA) in mice. METHODS: Mice were divided randomly into six groups, including normal, CIA, CIA+GCS (60, 120, and 240 mg/kg) and CIA plus glucosides of Tripterygium wilfordii (GTW) groups. CIA model was based on mice. The effect of GCS in CIA mice was measured by paw swelling, arthritis scores, and histopathological assessment of synovium. Indices of thymus and spleens were measured. Thymocytes and splenocytes proliferation, activity of interleukin-1 (IL-1), and interleukin-2 (IL-2) were assayed by MTT and [(3)H]TdR method. The level of anti-collagen type II (CII) antibody in serum and prostaglandin E (PGE) in ankle were assayed by ELISA and ultraviolet spectrophotometer method, respectively. RESULTS: The onset of paw-swelling was on d 24 after injection of emulsion. The peak of secondary inflammation appeared on d 36 and then declined after d 40. GCS and GTW significantly reduced paw-swelling and arthritis scores, reduced the increase of spleen indices of CIA mice, suppressed the ConA or LPS-induced thymocyte or spleen cell proliferation, and the production of IL-1 and IL-2 in CIA mice. GCS reduced the level of anti-CII antibody and PGE. Histological pathology analysis demonstrated that the synovium of CIA mice was hyperplastic, pannus was formed, and inflammatory cells infiltrated into synovium. The pathological changes were significantly reduced by GCS. CONCLUSION: GCS had anti-inflammatory effect on CIA mice, which might be related to the modification of the abnormal immunological function of CIA mice. PMID- 15525474 TI - Norcantharidin induces apoptosis in HeLa cells through caspase, MAPK, and mitochondrial pathways. AB - AIM: To investigate the mechanism of norcantharidin (NCTD)-induced HeLa cell apoptosis. METHODS: HeLa cell growth inhibition was measured by MTT method. Apoptosis was detected by Hoechst 33258 staining and agarose gel electrophoresis. Caspase activities were assayed using caspase apoptosis detection kit. Western blot analysis was used to evaluate the level of ICAD, ERK/p-ERK, JNK/p-JNK, and Bcl-X(L)/Bax expression. RESULTS: Norcantharidin inhibited HeLa cell growth in a time- and dose-dependent manner. HeLa cells treated with norcantharidin showed typical characteristics of apoptosis including the morphological changes and DNA fragmentation. Caspase family inhibitor (z-VAD-fmk), caspase-8, -9 inhibitor (z IETD-fmk, Ac-LEHD-CHO, respectively) and caspase-3 inhibitor (z-DEVD-fmk) partially prevent norcantharidin-induced apoptosis, but initiator caspase-1 inhibitor (Ac-YVAD-fmk) did not. The activities of caspase-3, -8, and -9 were up regulated after norcantharidin treatment. Furthermore, NCTD-induced activation of caspase-3 resulted in the degradation of the inhibitor of caspase-activated DNase (ICAD). Up-regulation of mitochondrial Bax expression and down-regulation of Bcl x(L) expression also participated in the apoptosis induced by NCTD. Although p38 MAPK inhibitor (SB203580) failed to block cell death, ERK MAPK inhibitor (PD98059) and JNK MAPK inhibitor (SP600125) had marked inhibitory effects on norcantharidin-induced apoptosis. Moreover, the phosphorylation of JNK were up regulated followed by delayed ERK phosphorylation after treatment with NCTD, suggesting that ERK and JNK were both responsible for NCTD-induced apoptosis in HeLa cells and worked at different stages. CONCLUSION: The cytotoxic effect of NCTD on HeLa cells was mainly due to apoptosis. The anti-tumor mechanism of NCTD might involve caspses, mitochondrial, and MAPKs pathways. PMID- 15525475 TI - PC-407 inhibited proliferation and induced apoptosis in human colon cancer SW 1116 cells. AB - AIM: To study whether PC-407 [4-[5-naphthyl-3- (trifluoromethyl)-1H-pyrazol-1-yl] benzenesulfonamide] inhibits cell viability and induces apoptosis in human colon cancer SW-1116 cells. METHODS: Inhibition of SW-1116 proliferation was measured by MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope and electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. The amount of apoptotic cells was measured by flow cytometry. RESULTS: PC-407 inhibited SW-1116 cell proliferation in a concentration-dependent manner after 3 d of treatment, and the IC(50) for PC-407 inhibition of cell number was 16.67+/-0.17 micromol/L. After incubation of SW 1116 cells with PC-407 20 micromol/L for 24 h, morphological changes of typical apoptosis were observed by AO/EB staining or transmission electron microscopy. Flow cytometry analysis showed that PC-407 induced apoptosis in SW-1116 cells in a time- and concentration-dependent manner. The agarose gel electrophoresis of DNA revealed a ladder pattern 48 h later. CONCLUSION: PC-407 inhibited proliferation and induced apoptosis in the human colon cancer SW-1116 cell line. PMID- 15525476 TI - 2-Methoxyestradiol induces cell cycle arrest and apoptosis of nasopharyngeal carcinoma cells. AB - AIM: To investigate 2-methoxyestradiol induced apoptosis and its mechanism of action in CNE2 cell lines. METHODS: CNE2 cells were cultured in RPMI-1640 medium and treated with 2-methoxyestradiol in different concentrations. MTT assay was used to detect growth inhibition. Flow cytometry and DNA ladders were used to detect apoptosis. Western blotting was used to observe the expression of p53, p21(WAF1), Bax, and Bcl-2 protein. RESULTS: 2-methoxyestradiol inhibited proliferation of nasopharyngeal carcinoma CNE2 cells with IC(50) value of 2.82 micromol/L. The results of flow cytometry showed an accumulation of CNE2 cells in G2/M phase in response to 2-methoxyestradiol. Treatment of CNE2 cells with 2 methoxyestradiol resulted in DNA fragmentation. The expression levels of protein p53 and Bcl-2 decreased following 2-methoxyestradiol treatment in CNE2 cells, whereas Bax and p21(WAF1) protein expression were unaffected after treatment with 2-methoxyestradiol. CONCLUSION: These results suggest that 2-methoxyestradiol induced cell cycle arrest at G2/M phase and apoptosis of CNE2 cells which was associated to Bcl-2 down-regulation. PMID- 15525478 TI - Proceedings of the 5th china-French pharmacological joint symposium. PMID- 15525477 TI - Structure-activity relationship of lysophosphatidylcholines in HL-60 human leukemia cells. AB - AIM: To explore the structure-activity relationship of lysophosphatidylcholine (LPC) and lysolipid molecules from a marine sponge and ladybirds. METHODS: We tested three synthetic LPCs and four natural lysolipids on Ca2+ mobilization in HL-60 human leukemia cells. RESULTS: We observed lysolipid-mediated Ca2+ mobilization. The activity was the same in both ester- and ether-linked lysolipids, and introduction of a double bond or methoxy group on the alkyl chain did not significantly modulate the activity. However, replacement of trimethylammonium moiety in the choline structure with ammonium moiety reduced the activity. Furthermore, change of the alkyl chain length influenced the Ca2+ response. CONCLUSION: LPC-induced Ca2+ mobilization might be dependent on the length of alkyl chain and the presence of choline moiety in HL-60 leukemia cells. PMID- 15525479 TI - Identification and assessment of ongoing trials in health technology assessment reviews. AB - OBJECTIVES: To assess the importance of ongoing trials in health technology assessment reviews (HTARs) for the National Institute for Clinical Excellence and to provide practical recommendations for identifying ongoing trials and assessing their possible impact. DATA SOURCES: Electronic databases. REVIEW METHODS: Ongoing trials (or trials in progress) were defined as any trials that have started but where the results are not yet available or only interim results are available for HTARs. This methodological review included: (1) an assessment of ongoing trials in HTARs completed by the end of August 2002, (2) a survey and assessment of trial registers and other sources of ongoing trials and (3) a summary and assessment of available methods for assessing the possible impacts of ongoing trials. RESULTS: The identification of ongoing trials is a common phenomenon in reviews of health technology assessment. Twenty-three of the 32 HTARs identified one or more ongoing trials and in eight of these the information on identified ongoing trials was not considered in the evidence synthesis and research recommendations. All but one HTAR that considered the potential impact of ongoing trials adopted a narrative approach. Trial registers and grey literature are important sources of information on ongoing trials. All 32 HTARs explicitly or implicitly searched for unpublished studies, and/or ongoing trials and/or grey literature and trial registers. The assessment of six commonly used trial registers suggested that most registers provided sufficient information for reviewers to decide the relevance of identified ongoing trials. However, it is sometimes extremely difficult to know whether ongoing trials identified from different sources (registers) are the same trials or belong to the same multicentre trials. The ISRCTN (the International Standard Randomised Controlled Trial Number) is the most reliable system but it has not been widely adopted. The qualitative assessment of ongoing trials compared major features of completed and ongoing trials, providing information about the possible impact of ongoing trials in terms of relevance, validity, reliability and generalisability. Quantitative methods to assess the impact of ongoing trials include cumulative meta-analysis related methods, fail-safe N, Bayesian data monitoring, and Bayesian interim predictions. The most useful method may be the Bayesian predictive probability, which estimates predictive probabilities for any possible values of treatment effect. A case study indicated that the appropriate use of quantitative methods would strengthen findings from narrative assessment of possible impact of ongoing trials. CONCLUSIONS: Identification of ongoing trials is common in HTARs. Searching for ongoing trials in effectiveness reviews should be more thorough and explicit. Conversely, primary researchers, in particular those working with in multicentre trials, should label ongoing trials more clearly, preferably by ISRCTN. Qualitative assessment of identified ongoing trials is crucial and informative. Available quantitative methods could be used to strengthen findings from narrative assessment, although further research and more empirical examples are required. Information from ongoing trials may contribute to syntheses of results, conclusions and recommendations for future research. Future research is suggested into the identification and assessment of ongoing trials in other systematic reviews of effectiveness of health care interventions; existing and new methods for incorporating information on ongoing trials; comparing estimated impacts with the actual results of ongoing trials; and to incorporate findings from the assessment of ongoing trials into decision models. PMID- 15525481 TI - [Receptors, receptors...]. PMID- 15525480 TI - Systematic review and economic evaluation of a long-acting insulin analogue, insulin glargine. AB - OBJECTIVES: To evaluate the use of insulin glargine in its licensed basal-bolus indication in terms of both clinical and cost-effectiveness. DATA SOURCES: Electronic databases. REVIEW METHODS: A systematic review of the literature, involving a range of databases, was performed to identify all papers relating to insulin glargine. RESULTS: Nineteen studies met the inclusion criteria but full reports were available for only six. For type 1 diabetes patients, insulin glargine appears to be more effective than neutral protamine Hagedorn (NPH) in reducing fasting blood glucose (FBG) but not in reducing glycosylated haemoglobin (HbA1c) and there is some evidence that both insulins are as effective as each other in both FBG and HbA1c control. For type 2 patients for whom oral antidiabetic agents provide inadequate glycaemic control, there is no evidence that insulin glargine is more effective than NPH in reducing either FBG or HbA1c and some evidence that both insulins are as effective as each other in both FBG and HbA1c control. Evidence for control of hypoglycaemia is equivocal. In studies where insulin glargine is demonstrated to be superior to NPH in controlling nocturnal hypoglycaemia, this may be only apparent when compared with once-daily NPH and not twice-daily NPH. Further, this superiority of glargine over NPH in the control of nocturnal hypoglycaemia may relate to one formulation of insulin glargine (HOE901[80]) and not another (HOE901[30]). There is no conclusive evidence that insulin glargine is superior to NPH in controlling symptomatic hypoglycaemia and severe hypoglycaemia. Insufficient data are available to conclude whether insulin glargine is different from each of the commonly used NPH dosing regimens: once daily and more than once daily. Given the lack of a published evidence base for the cost-effectiveness of insulin glargine, the economic review concentrates on a review of the industry submission and an amended model. Three economic models are provided in the submission, two relating to type 1 diabetes and one relating to type 2 diabetes. All three models compare the cost--utility of insulin glargine against NPH insulin. In general, the structures of the models are poor and in all three models, mistakes relating to assumptions and calculations have been made. The assessment team believe that the cost per QALY estimates generated by the Aventis model may be an underestimate for several reasons. The cost-effectiveness of insulin glargine in both type 1 and type 2 diabetes is highly sensitive to the amount of utility associated with reducing the fear of hypoglycaemia. CONCLUSIONS: The evidence suggests that, compared with NPH insulin, insulin glargine is effective in reducing the number of nocturnal hypoglycaemic episodes, especially when compared with once-daily NPH. There appears to be no improvement in long-term glycaemic control and therefore insulin glargine is unlikely to reduce the incidence of the long-term microvascular and cardiovascular complications of diabetes. Further research into insulin glargine is needed that addresses the quality of life issues associated with fear of hypoglycaemia and also the economic impact of balance of HbA1c control and incidence of hypoglycaemia achieved in practice. Studies examining the economic evidence on insulin glargine should be published. PMID- 15525483 TI - [RGM and neogenin, a promising couple]. PMID- 15525484 TI - [Stimulation of BDNF transport by huntingtin]. PMID- 15525485 TI - [Cornelia de Lange syndrome]. PMID- 15525486 TI - [A novel mechanism for genetic diseases: transcription of antisense RNA may silence a gene]. PMID- 15525487 TI - [Wired for eating]. PMID- 15525488 TI - [HOXB4 and primitive human hematopoietic cell expansion]. PMID- 15525489 TI - [Fli1 haploinsufficiency underlies Paris-Trousseau thrombopenia]. PMID- 15525490 TI - [Pharmacogenomics of the growth hormone: polymorphism of the receptor in the front line]. PMID- 15525492 TI - [Clinical use of calcimimetics in the treatment of hyperparathyroidisms]. AB - Recognition of the role of the extracellular calcium sensing receptor (CaR) in mineral metabolism has greatly improved our understanding of calcium homeostasis. The activation of this receptor by small changes in the extracellular ionized calcium concentration (Ca(2+)ec) regulates parathormone (PTH) and calcitonin secretion, urinary calcium excretion and ultimately bone turnover. Cloning of CaR and discovery of mutations making the receptor less or more sensitive to calcium allowed a better understanding of several hereditary disorders characterized either by hyperparathyroidism or hypoparathyroidism. CaR became an ideal target for the development of compounds able to modulate the activity of CaR, activators (calcimimetics) as well as inhibitors (calcilytics). The calcimimetics are able to amplify the sensitivity of the CaR to Ca(2+)ec, suppressing PTH levels with a resultant fall in blood Ca2+. They dose-dependently reduce the secretion of PTH in vitro in cultured parathyroid cells, in animal models and in humans. In uremic animals, these compounds prevent parathyroid cell hyperplasia, normalize plasma PTH levels and bone remodelling. In uremic patients undergoing hemodialysis, the calcimimetics reduce plasma PTH concentration at short-term (12 weeks) as well as at long-term (2 years), serum calcium-phosphorus product and bone remodelling. After one year of treatment, these patients show a gain of bone mass of 2-3% at the femoral neck and at the total body. Contrarily, the calcilytics, by inhibiting CaR, can intermittently stimulate the secretion and the serum concentration of PTH. This results in an skeletal anabolic effect with a substantial increase in bone mineral density. They are potentially very interesting for the treatment of post-menopausal osteoporosis. PMID- 15525493 TI - [New insights into the pharmacology of the extracellular calcium sensing receptor]. AB - The extracellular calcium-sensing receptor (CaR) belongs to class III of G protein coupled receptors. The CaR is expressed at the surface of the parathyroid cells and plays an essential role in the regulation of Ca2+ homeostasis through the control of parathyroid secretion. The CaR is activated by Ca2+ and Mg2+ present in the extracellular fluids, various di- and trivalent cations, L aminoacids and charged molecules including several antibiotics. Calcimimetics potentiate the effect of Ca2+ and are proposed to be of therapeutic benefit for the treatment of both primary and secondary hyperparathyroidism. Calcilytics block the Ca2+-induced activation of the CaR. Three-dimensional models of the seven transmembrane domains of the human CaR have been used to identify specific residues implicated in the recognition of calcimimetics and calcilytics. These molecules should be useful for delineating the physiological roles played by the CaR in several tissues and for clarifying the direct effects attributed to extracellular Ca2+. PMID- 15525494 TI - [Biotechnological and administration innovations in insulin therapy]. AB - The importance of the intensive control of blood glucose in patients with diabetes has been well documented in several large scale studies. Attempts to attain strict glucose control when managing diabetes have traditionally utilized daily subcutaneous injections of human insulin. This strategy has offered improvements in glycaemic control but is unable to replicate fully the normal diurnal plasma profile of endogenous human insulin. Advances in protein engineering techniques have, however, resulted in the formulation of a number of insulin analogues that offer more desirable properties of absorption from the subcutaneous depot and hence improved insulin profiles in patients with diabetes. Concurrent to the development of insulin analogues, devices to deliver insulin either subcutaneously or by other routes have also advanced. These novel delivery strategies are also likely to contribute to improved glycaemic control for patients with diabetes in the future. PMID- 15525495 TI - [Immunoprophylaxis of respiratory infections]. AB - Anti-infective antibody-based immunotherapy has gained renewed interest since the crisis of antibiotic resistance and because there is no therapy against various viral infections. The immunoprophylaxis of respiratory infections aims to utilize the ability of local antibodies to neutralize inhaled micro-organisms and their cytopathic products. Immunoglobulins for intravenous use (i.v.i.g.) have a wide spectrum of specificities. Hyperimmune i.v.i.g. containing high titers of specific antibodies have demonstrated efficacy in clinical trials, notably against the respiratory syncytial virus. Monoclonal antibodies have the advantage to be homogenous and specific for one selected epitope and several studies have demonstrated their efficacy to neutralize several infectious agents. Moreover, antibodies can be administered topically and are effective at lower doses than those needed for systemic administration. The mechanism of action could be the agglutination of bacteria or viruses at the epithelial surfaces of the respiratory tract inhibiting the early steps of the infectious process. Thanks to new technologies of humanized monoclonal antibodies, immunotherapy offers real promising perspectives for prophylactic and therapeutic therapies against a variety of current or emerging infectious diseases. PMID- 15525496 TI - [Bird sex determination]. AB - During the evolution, sex determination occurred early. Sex determining factors were progressively isolated from other genes in sexual chromosomes, or gonosomes. Among vertebrates, evolution took two opposite pathways : in mammals, the system of XX:XY sex determination, with Y chromosome, induces male differentiation. In contrast, in birds, the system ZZ:ZW, with the W chromosome, induces female differentiation. But comparative studies show that the two pathways are not so simple. In the chicken as in the lower vertebrates, estrogens play a central role in gonadal sex differentiation. Several genes, show to be critical for mammalian determination, are also expressed in the chicken but their expression pattern differs, indicating functional plasticity. The W-linked female determinants remains still unknown. But comparative studies of the two pathways, with conserved and divergent elements, are broadening our understanding of sex determination. PMID- 15525497 TI - [Tpit mutations reveal a new model of pituitary differentiation and account for isolated ACTH deficiency]. AB - Pituitary hormone-producing cells differentiate sequentially from a common epithelial primordium, Rathke's pouch, under the combinatorial action of a subset of tissue- and cell-restricted transcription factors. Some factors have been implicated in early events of pituitary induction and morphogenesis while other factors like Pit-1 and SF-1 have been associated with differentiation of particular lineages. In POMC-expressing cells, Pitx1, NeuroD1 and Tpit were shown to be important for cell specific transcription of the POMC gene. Since Tpit is exclusively expressed in pituitary POMC-expressing lineages, the corticotrophs and melanotrophs, we investigated the TPIT gene coding sequences in 17 patients presenting with congenital isolated ACTH deficiency (IAD). We demonstrated that human TPIT gene mutations cause a neonatal onset form of IAD (8/11), but not juvenile forms of this deficiency (0/6). In the absence of glucocorticoid replacement, IAD can lead to neonatal death by acute adrenal insufficiency. To assess the importance of Tpit in pituitary differentiation and function, we produced Tpit-null mice. Concordant with the human phenotype, Tpit-null mice have IAD : plasma ACTH is greatly reduced in these mice, their plasma corticosterone is undetectable and the adrenals are hypoplastic. Analysis of the pituitary in Tpit-null mice revealed multiple roles of this factor in cell differentiation. First, Tpit is a positive regulator for POMC cell differentiation. Tpit is also a negative regulator of the pituitary gonadotroph fate. Thus, Tpit operates as a molecular switch to orient differentiation of a common precursor towards either POMC or gonadotroph fate. A binary choice model of pituitary cell differentiation is presented. PMID- 15525498 TI - [Oncogenic transcription factors as splicing regulators]. AB - Oncogene activity ranges from transduction signals to transcription factors. Altered expression of oncogenes, either by chromosomal translocation, proviral insertion or point mutations, can lead to tumor formation. More specifically, data accumulated through the last two decades have shown that disregulation of oncogenic transcription factors can interfere with regulatory cascades that control the growth, differentiation, and survival of normal cells. There is also evidence that alterations of oncogene activity are associated with pre-mRNA splicing defects. The insights gained from the pivotal role of RNA polymerase II in coupling transcription and splicing have instigated a new line of research regarding the possible role of oncogenic transcription factors in pre-mRNA splicing regulation. This review focuses on recent advances addressing this question. Understanding the impact of alterations in the expression and/or function of oncogenes have important prognostic implications that can guide the design of new therapeutic drugs to promote differentiation and/or apoptosis over cell proliferation. PMID- 15525499 TI - [The double-play of PP17/TIP47]. AB - A missing link in the understanding of the mechanisms of transport of the mannose 6-phosphate receptors has recently been discovered, following the identification of the protein TIP47. In association with Rab9-GTP, this protein is responsible for the return of the receptors from the late endosomes back to the trans-Golgi network. Curiously, the same protein called PP17b, was described as a placental protein twenty years ago, and more recently, as a blood marker for human uterine cervical cancer. The sequence of PP17b/TIP47 displays not only a strong homology with those of adipophilin and the perilipins, two proteins known to be involved in the intracellular traffic of lipid droplets but also PP17b/TIP47 is associated with the later. How this ubiquitous protein could participate in processes as different as the mannose 6-phosphate receptors traffic and the formation and/or traffic of lipid droplets? A tentative hypothesis is put forward. PMID- 15525500 TI - [FRAP, FLIP, FRET, BRET, FLIM, PRIM...new techniques for a colourful life]. AB - Cell and tissue imaging provides scientists with wonderful tools, thanks to a fruitful dialog between chemistry, optical, mechanical, computational sciences and biology. Confocal microscopy, videomicroscopy together with a new generation of fluorochromes (especially those derived from green fluorescent protein, GFP) and image analysis software allow to visualize life in all its dimensions (space and time). Cell imaging also allows to quantify biological processes at the cellular level, to analyse both stoechiometry and dynamics of molecular interactions involved in cell and tissue regulations. Entering the new era of post-genomics requires a better knowledge of advantages and limitations of these new approaches. PMID- 15525501 TI - [Prediction of transcription and genomic sequences]. AB - Technological developments have enhanced DNA sequencing at genomic scale. On the basis of the resulting sequences, computational biologists now attempt to localise the most important functional regions, starting with genes, but also importantly the regulatory motifs and conditions controlling their expression. In a recent paper published in Cell, M.A. Beer and S. Tavazoie report the results obtained by combining statistical classifications (clustering) of transcriptome data (DNA chips), software for the discovery of cis-regulatory patterns, together with a probabilistic learning method to infer regulatory rules tentatively accounting for the observed transcriptional profiles. PMID- 15525502 TI - [Eggs in plenty?]. PMID- 15525503 TI - [Nobel Prize 2004 for Medicine. Richard Axel and Linda B. Buck. Olfaction under the spotlight]. PMID- 15525504 TI - [Social withdrawal behaviour in infant: clinical and research interest]. AB - The social withdrawal behaviour concept is presented, along with its historical background and its links to infant depression. Sustained social withdrawal behaviour in infancy is observed either with attachment disorders, in autism, sensory impairment, intense and chronic pain, or in interaction disorders and as effects of maternal depression on the infant. The alarm distress baby scale (ADBB) is a 8 item-assessment scale. It has been built to help screening early withdrawal behaviour, making use for example of the routine physical examination in a Well Baby clinic. Current clinical and research application and validations of the scale are presented. PMID- 15525505 TI - [An enigmatic disease in Gregor Mendel's life]. AB - The great value of the experimental and theoretical work of Gregor Mendel has been recognized more than thirty five years after its publication; in this article, we suggest that his personality has still to be rediscovered. PMID- 15525506 TI - Importin beta: conducting a much larger cellular symphony. AB - Importin beta, once thought to be exclusively a nuclear transport receptor, is emerging as a global regulator of diverse cellular functions. Importin beta acts positively in multiple interphase roles: in nuclear import, as a chaperone for highly charged nuclear proteins, and as a potential motor adaptor for movement along microtubules. In contrast, importin beta plays a negative regulatory role in mitotic spindle assembly, centrosome dynamics, nuclear membrane formation, and nuclear pore assembly. In most of these, importin beta is counteracted by its regulator, Ran-GTP. In light of this, the recent discovery of Ran's involvement in spindle checkpoint control suggested a potential new arena for importin beta action, although it is also possible that one of importin beta's relatives, the karyopherin family of proteins, manages this checkpoint. Lastly, importin beta plays a role in transducing damage signals from the axons of injured neurons back to the cell body. PMID- 15525507 TI - NaBC1 is a ubiquitous electrogenic Na+ -coupled borate transporter essential for cellular boron homeostasis and cell growth and proliferation. AB - Boron is a vital micronutrient in plants and may be essential for animal growth and development. Whereas the role of boron in the life cycle of plants is well documented, nothing is known about boron homeostasis and function in animal cells. NaBC1, the mammalian homolog of AtBor1, is a borate transporter. In the absence of borate, NaBC1 conducts Na(+) and OH(-) (H(+)), while in the presence of borate, NaBC1 functions as an electrogenic, voltage-regulated, Na(+)-coupled B(OH)(4)(-) transporter. At low concentrations, borate activated the MAPK pathway to stimulate cell growth and proliferation, and at high concentrations, it was toxic. Accordingly, overexpression of NaBC1 shifted both effects of borate to the left, whereas knockdown of NaBC1 halted cell growth and proliferation. These findings may reveal a previously unrecognized role for NaBC1 in borate homeostasis and open the way to better understanding of the many presumed physiological roles of borate in animals. PMID- 15525508 TI - Nucleotide-dependent substrate handoff from the SspB adaptor to the AAA+ ClpXP protease. AB - The SspB adaptor enhances ClpXP degradation by binding the ssrA degradation tag of substrates and the AAA+ ClpX unfoldase. To probe the mechanism of substrate delivery, we engineered a disulfide bond between the ssrA tag and SspB and demonstrated otherwise normal interactions by solving the crystal structure. Although the covalent link prevents adaptor.substrate dissociation, ClpXP degraded GFP-ssrA that was disulfide bonded to the adaptor. Thus, crosslinked substrate must be handed directly from SspB to ClpX. The ssrA tag in the covalent adaptor complex interacted with ClpX.ATPgammaS but not ClpX.ADP, suggesting that handoff occurs in the ATP bound enzyme. By contrast, SspB alone bound ClpX in both nucleotide states. Similar handoff mechanisms will undoubtedly be used by many AAA+ adaptors and enzymes, allowing assembly of delivery complexes in either nucleotide state, engagement of the recognition tag in the ATP state, and application of an unfolding force to the attached protein following hydrolysis. PMID- 15525509 TI - Structure of the Tetrahymena ribozyme: base triple sandwich and metal ion at the active site. AB - The Tetrahymena intron is an RNA catalyst, or ribozyme. As part of its self splicing reaction, this ribozyme catalyzes phosphoryl transfer between guanosine and a substrate RNA strand. Here we report the refined crystal structure of an active Tetrahymena ribozyme in the absence of its RNA substrate at 3.8 A resolution. The 3'-terminal guanosine (omegaG), which serves as the attacking group for RNA cleavage, forms a coplanar base triple with the G264-C311 base pair, and this base triple is sandwiched by three other base triples. In addition, a metal ion is present in the active site, contacting or positioned close to the ribose of the omegaG and five phosphates. All of these phosphates have been shown to be important for catalysis. Therefore, we provide a picture of how the ribozyme active site positions both a catalytic metal ion and the nucleophilic guanosine for catalysis prior to binding its RNA substrate. PMID- 15525510 TI - A pathway of sequential arginine-serine-rich domain-splicing signal interactions during mammalian spliceosome assembly. AB - Serine-arginine (SR) proteins are general splicing factors and can function through binding to exonic splicing enhancers (ESEs). SR proteins and several other mammalian splicing factors contain an arginine-serine-rich (RS) domain required to promote splicing. We have recently found that the ESE bound RS domain functions by contacting the branchpoint. Here, we perform RNA-protein crosslinking experiments to show that the branchpoint is sequentially contacted first in complex E by the RS domain of the essential splicing factor U2AF(65) and then in the prespliceosome by the ESE bound RS domain. Although the ESE bound RS domain can promote formation of the prespliceosome, at least one additional SR protein is required for complete spliceosome assembly. We show that the RS domain of this additional SR protein contacts the 5' splice site specifically in the mature spliceosome. We propose that direct contact with splicing signals is a general mechanism by which RS domains promote splicing. PMID- 15525511 TI - Achieving error-free translation; the mechanism of proofreading of threonyl-tRNA synthetase at atomic resolution. AB - The fidelity of aminoacylation of tRNA(Thr) by the threonyl-tRNA synthetase (ThrRS) requires the discrimination of the cognate substrate threonine from the noncognate serine. Misacylation by serine is corrected in a proofreading or editing step. An editing site has been located 39 A away from the aminoacylation site. We report the crystal structures of this editing domain in its apo form and in complex with the serine product, and with two nonhydrolyzable analogs of potential substrates: the terminal tRNA adenosine charged with serine, and seryl adenylate. The structures show how serine is recognized, and threonine rejected, and provide the structural basis for the editing mechanism, a water-mediated hydrolysis of the mischarged tRNA. When the adenylate analog binds in the editing site, a phosphate oxygen takes the place of one of the catalytic water molecules, thereby blocking the reaction. This rules out a correction mechanism that would occur before the binding of the amino acid on the tRNA. PMID- 15525512 TI - Phosphorylation of Cdc20 by Bub1 provides a catalytic mechanism for APC/C inhibition by the spindle checkpoint. AB - To ensure the fidelity of chromosome segregation, the spindle checkpoint blocks the ubiquitin ligase activity of APC/C(Cdc20) in response to a single chromatid not properly attached to the mitotic spindle. Here we show that HeLa cells depleted for Bub1 by RNA interference are defective in checkpoint signaling. Bub1 directly phosphorylates Cdc20 in vitro and inhibits the ubiquitin ligase activity of APC/C(Cdc20) catalytically. A Cdc20 mutant with all six Bub1 phosphorylation sites removed is refractory to Bub1-mediated phosphorylation and inhibition in vitro. Upon checkpoint activation, Bub1 itself is hyperphosphorylated and its kinase activity toward Cdc20 is stimulated. Ectopic expression of the nonphosphorylatable Cdc20 mutant allows HeLa cells to escape from mitosis in the presence of spindle damage. Therefore, Bub1-mediated phosphorylation of Cdc20 is required for proper checkpoint signaling. We speculate that inhibition of APC/C(Cdc20) by Bub1 in a catalytic fashion may partly account for the exquisite sensitivity of the spindle checkpoint. PMID- 15525513 TI - A common set of gene regulatory networks links metabolism and growth inhibition. AB - Using genome-wide analysis of transcription factor occupancy, we investigated the mechanisms underlying three mammalian growth arrest pathways that require the pRB tumor suppressor family. We found that p130 and E2F4 cooperatively repress a common set of genes under each growth arrest condition and showed that growth arrest is achieved through repression of a core set of genes involved not only in cell cycle control but also mitochondrial biogenesis and metabolism. Motif finding algorithms predicted the existence of nuclear respiratory factor-1 (NRF1) binding sites in E2F target promoters, and genome-wide factor binding analysis confirmed our predictions. We showed that NRF1, a factor known to regulate expression of genes involved in mitochondrial function, is a coregulator of a large number of E2F target genes. Our studies provide insights into E2F regulatory circuitry, suggest how factor occupancy can predict the expression signature of a given target gene, and reveal pathways deregulated in human tumors. PMID- 15525514 TI - p53 linear diffusion along DNA requires its C terminus. AB - In cells, sequence-specific transcription factors must search through an entire genome to find their target sites in promoters. Such sites may be identified by using one-dimensional (linear diffusion) and/or three-dimensional (association/dissociation) mechanisms. We show here that wild-type p53 possesses the ability to linearly diffuse on DNA. p53 lacking its C terminus is incapable of such sliding along DNA, while the isolated C terminus of p53 is even more effective than the full-length protein at one-dimensional linear diffusion. Importantly, neither acetylation-mimicking mutations nor phosphorylation of residues within the C terminus stimulates linear diffusion by p53. Supporting these in vitro observations, we found that C-terminally deleted p53 (p53Delta30) expressed at physiological levels is deficient in binding to and transactivating downstream promoters in vivo. Therefore, our data show that the C terminus is a positive regulator of DNA binding in vivo and in vitro, and indicate that the mechanism may involve linear diffusion. PMID- 15525515 TI - Structural basis for androgen receptor interdomain and coactivator interactions suggests a transition in nuclear receptor activation function dominance. AB - The androgen receptor (AR) is required for male sex development and contributes to prostate cancer cell survival. In contrast to other nuclear receptors that bind the LXXLL motifs of coactivators, the AR ligand binding domain is preferentially engaged in an interdomain interaction with the AR FXXLF motif. Reported here are crystal structures of the ligand-activated AR ligand binding domain with and without bound FXXLF and LXXLL peptides. Key residues that establish motif binding specificity are identified through comparative structure function and mutagenesis studies. A mechanism in prostate cancer is suggested by a functional AR mutation at a specificity-determining residue that recovers coactivator LXXLL motif binding. An activation function transition hypothesis is proposed in which an evolutionary decline in LXXLL motif binding parallels expansion and functional dominance of the NH(2)-terminal transactivation domain in the steroid receptor subfamily. PMID- 15525516 TI - DNA instructed displacement of histones H2A and H2B at an inducible promoter. AB - Regulation of gene expression requires dynamic changes in chromatin, but the nature of these changes is not well understood. Here, we show that progesterone treatment of cultured cells leads to recruitment of progesterone receptor (PR) and SWI/SNF-related complexes to Mouse Mammary Tumor Virus (MMTV) promoter, accompanied by displacement of histones H2A and H2B from the nucleosome containing the receptor binding sites, but not from adjacent nucleosomes. PR recruits SWI/SNF to MMTV nucleosomes in vitro and facilitates synergistic binding of receptors and nuclear factor 1 to the promoter. In nucleosomes assembled on MMTV or mouse rDNA promoter sequences, SWI/SNF catalyzes ATP-dependent sliding of the histone octamer followed only on the MMTV promoter by displacement of histones H2A and H2B. In MMTV nucleosome arrays, SWI/SNF displaces H2A and H2B from nucleosome B and not from the adjacent nucleosome. Thus, the outcome of nucleosome remodeling by SWI/SNF depends on DNA sequence. PMID- 15525517 TI - Recruitment of histone modifications by USF proteins at a vertebrate barrier element. AB - The chicken beta-globin 5'HS4 insulator element acts as a barrier to the encroachment of chromosomal silencing. Endogenous 5'HS4 sequences are highly enriched with histone acetylation and H3K4 methylation regardless of neighboring gene expression. We report here that 5'HS4 elements recruit these histone modifications when protecting a reporter transgene from chromosomal silencing. Deletion studies identified a single protein binding site within 5'HS4, footprint IV, that is necessary for the recruitment of histone modifications and for barrier activity. We have determined that USF proteins bind to footprint IV. USF1 is present in complexes with histone modifying enzymes in cell extracts, and these enzymes specifically interact with the endogenous 5'HS4 element. Knockdown of USF1 expression leads to a loss of histone modification recruitment and subsequent encroachment of H3K9 methylation. We propose that barrier activity requires the constitutive recruitment of H3K4 methylation and histone acetylation at multiple residues to counteract the propagation of condensed chromatin structures. PMID- 15525518 TI - Rvb1p/Rvb2p recruit Arp5p and assemble a functional Ino80 chromatin remodeling complex. AB - The Rvb1p and Rvb2p (or TIP48 and TIP49) nuclear ATP binding proteins are universally conserved in eukaryotes and essential for viability of yeasts. Rvbp associate with each other as a double hexamer, with YHR034c and with two complexes involved in chromatin remodeling, Ino80.com and Swr1.com. Loss of Rvb1p or Ino80p affects many yeast promoters similarly. Rvbp are not essential for the recruitment of Ino80p to promoters but are essential for the catalytic activity of Ino80.com. Loss of Rvbp leads to loss of the functionally critical Arp5p in Ino80.com. Rvb2p associates with Arp5p in vitro in a reaction dependent on the presence of ATP and Ino80p. Therefore, Rvbp are required for the structural and functional integrity of the Ino80 chromatin remodeling complex. PMID- 15525519 TI - The INO80 protein controls homologous recombination in Arabidopsis thaliana. AB - Homologous recombination (HR) serves a dual role in providing genetic flexibility and in maintaining genome integrity. Little is known about the regulation of HR and other repair pathways in the context of chromatin. We report on a mutant affected in the expression of the Arabidopsis INO80 ortholog of the SWI/SNF ATPase family, which shows a reduction of the HR frequency to 15% of that in wild type plants. In contrast, sensitivity to genotoxic agents and efficiency of T-DNA integration remain unaffected, suggesting that INO80 is a positive regulator of HR, while not affecting other repair pathways. So far, INO80 function has only been reported in a lower eukaryote. Profiling studies on three ino80 allelic mutants show that INO80 regulates nearly 100 Arabidopsis genes. However, the transcriptional regulation of repair-related genes is unaffected in the mutant. This suggests a dual role for INO80 in transcription and DNA repair by HR. PMID- 15525520 TI - A robust toolkit for functional profiling of the yeast genome. AB - Study of mutant phenotypes is a fundamental method for understanding gene function. The construction of a near-complete collection of yeast knockouts (YKO) and the unique molecular barcodes (or TAGs) that identify each strain has enabled quantitative functional profiling of Saccharomyces cerevisiae. By using these TAGs and the SGA reporter, MFA1pr-HIS3, which facilitates conversion of heterozygous diploid YKO strains into haploid mutants, we have developed a set of highly efficient microarray-based techniques, collectively referred as dSLAM (diploid-based synthetic lethality analysis on microarrays), to probe genome-wide gene-chemical and gene-gene interactions. Direct comparison revealed that these techniques are more robust than existing methods in functional profiling of the yeast genome. Widespread application of these tools will elucidate a comprehensive yeast genetic network. PMID- 15525522 TI - MicroRNAs guide asymmetric DNA modifications guiding asymmetric organs. AB - In plants and animals, microRNAs have been shown to regulate target genes by inhibiting translation or altering target mRNA stability. In this issue of Developmental Cell, Bao et al. extend the known mechanisms of action of microRNAs to RNA-directed DNA methylation, a mechanism previously associated only with siRNA-mediated gene silencing. PMID- 15525523 TI - Mitochondrial programmed cell death pathways in yeast. AB - Whether or not yeast cell death is altruistic, apoptotic, or otherwise analogous to programmed cell death in mammals is controversial. However, growing attention to cell death mechanisms in yeast has produced several new papers that make a case for ancient origins of programmed death involving mitochondrial pathways conserved between yeast and mammals. PMID- 15525524 TI - Atonal points the way- protein-protein interactions and developmental biology. AB - Many cells maintain their state of determination long after the signals that induced it decay. In this issue of Developmental Cell, zur Lage and colleagues describe how certain cells sustain proneural gene expression through direct interactions between transcription factors. PMID- 15525525 TI - Bub1, a gatekeeper for Cdc20-dependent mitotic exit. AB - The mitotic spindle assembly checkpoint arrests cells at metaphase by suppressing Cdc20, a protein required to trigger ubiquitination and consequent degradation of cyclin B. New evidence from Tang et al. appearing in the November 5th issue of Molecular Cell finds that one of the checkpoint proteins, Bub1, specifically phosphorylates Cdc20 to suppress APC/C activation. PMID- 15525526 TI - Stuck in division or passing through: what happens when cells cannot satisfy the spindle assembly checkpoint. AB - Cells that cannot satisfy the spindle assembly checkpoint (SAC) are delayed in mitosis (D-mitosis), a fact that has useful clinical ramifications. However, this delay is seldom permanent, and in the presence of an active SAC most cells ultimately escape mitosis and enter the next G1 as tetraploid cells. This review defines and discusses the various factors that determine how long a cell remains in mitosis when it cannot satisfy the SAC and also discusses the cell's subsequent fate. PMID- 15525527 TI - MicroRNA binding sites in Arabidopsis class III HD-ZIP mRNAs are required for methylation of the template chromosome. AB - Dominant mutations in the Arabidopsis PHABULOSA (PHB) and PHAVOLUTA (PHV) transcription factor genes cause transformation of abaxial to adaxial leaf fates by altering a microRNA complementary site present in processed PHB and PHV mRNAs but not in the corresponding genomic DNA. phb-1d mutants accumulate excess PHB transcript throughout the leaf primordium, indicating defective regulation of PHB transcript synthesis and/or stability. We show that PHB and PHV coding sequences are heavily methylated downstream of the microRNA complementary site in most wild type plant cells and that methylation is reduced in phb-1d and phv-1d mutants. Decreased methylation is limited to the chromosome bearing the dominant mutant allele in phb-1d heterozygotes. Low levels of methylation are detected in wt PHB DNA isolated from undifferentiated tissues. These results suggest a model in which the microRNA interacts with nascent, newly processed PHB mRNA to alter chromatin of the corresponding PHB template DNA predominantly in differentiated cells. PMID- 15525528 TI - Polycomb group proteins Ring1A/B link ubiquitylation of histone H2A to heritable gene silencing and X inactivation. AB - In many higher organisms, 5%-15% of histone H2A is ubiquitylated at lysine 119 (uH2A). The function of this modification and the factors involved in its establishment, however, are unknown. Here we demonstrate that uH2A occurs on the inactive X chromosome in female mammals and that this correlates with recruitment of Polycomb group (PcG) proteins belonging to Polycomb repressor complex 1 (PRC1). Based on our observations, we tested the role of the PRC1 protein Ring1B and its closely related homolog Ring1A in H2A ubiquitylation. Analysis of Ring1B null embryonic stem (ES) cells revealed extensive depletion of global uH2A levels. On the inactive X chromosome, uH2A was maintained in Ring1A or Ring1B null cells, but not in double knockout cells, demonstrating an overlapping function for these proteins in development. These observations link H2A ubiquitylation, X inactivation, and PRC1 PcG function, suggesting an unanticipated and novel mechanism for chromatin-mediated heritable gene silencing. PMID- 15525529 TI - The APC tumor suppressor binds to C-terminal binding protein to divert nuclear beta-catenin from TCF. AB - Adenomatous polyposis coli (APC) is an important tumor suppressor in the colon. APC antagonizes the transcriptional activity of the Wnt effector beta-catenin by promoting its nuclear export and its proteasomal destruction in the cytoplasm. Here, we show that a third function of APC in antagonizing beta-catenin involves C-terminal binding protein (CtBP). APC is associated with CtBP in vivo and binds to CtBP in vitro through its conserved 15 amino acid repeats. Failure of this association results in elevated levels of beta-catenin/TCF complexes and of TCF mediated transcription. Notably, CtBP is neither associated with TCF in vivo nor does mutation of the CtBP binding motifs in TCF-4 alter its transcriptional activity. This questions the idea that CtBP is a direct corepressor of TCF. Our evidence indicates that APC is an adaptor between beta-catenin and CtBP and that CtBP lowers the availability of free nuclear beta-catenin for binding to TCF by sequestering APC/beta-catenin complexes. PMID- 15525530 TI - EGF receptor signaling triggers recruitment of Drosophila sense organ precursors by stimulating proneural gene autoregulation. AB - In Drosophila, commitment of a cell to a sense organ precursor (SOP) fate requires bHLH proneural transcription factor upregulation, a process that depends in most cases on the interplay of proneural gene autoregulation and inhibitory Notch signaling. A subset of SOPs are selected by a recruitment pathway involving EGFR signaling to ectodermal cells expressing the proneural gene atonal. We show that EGFR signaling drives recruitment by directly facilitating atonal autoregulation. Pointed, the transcription factor that mediates EGFR signaling, and Atonal protein itself bind cooperatively to adjacent conserved binding sites in an atonal enhancer. Recruitment is therefore contingent on the combined presence of Atonal protein (providing competence) and EGFR signaling (triggering recruitment). Thus, autoregulation is the nodal control point targeted by signaling. This exemplifies a simple and general mechanism for regulating the transition from competence to cell fate commitment whereby a cell signal directly targets the autoregulation of a selector gene. PMID- 15525531 TI - FBF-1 and FBF-2 regulate the size of the mitotic region in the C. elegans germline. AB - In the C. elegans germline, GLP-1/Notch signaling and two nearly identical RNA binding proteins, FBF-1 and FBF-2, promote proliferation. Here, we show that the fbf-1 and fbf-2 genes are largely redundant for promoting mitosis but that they have opposite roles in fine-tuning the size of the mitotic region. The mitotic region is smaller than normal in fbf-1 mutants but larger than normal in fbf-2 mutants. Consistent with gene-specific roles, fbf-2 expression is limited to the distal germline, while fbf-1 expression is broader. The fbf-2 gene, but apparently not fbf-1, is controlled by GLP-1/Notch signaling, and the abundance of FBF-1 and FBF-2 proteins is limited by reciprocal 3'UTR repression. We propose that the divergent fbf genes and their regulatory subnetwork enable a precise control over size of the mitotic region. Therefore, fbf-1 and fbf-2 provide a paradigm for how recently duplicated genes can diverge to fine-tune patterning during animal development. PMID- 15525532 TI - Activation of a signaling cascade by cytoskeleton stretch. AB - Cells sense and respond to mechanical force. However, the mechanisms of transduction of extracellular matrix (ECM) forces to biochemical signals are not known. After removing the cell membrane and soluble proteins by Triton X-100 extraction, we found that the remaining complex (Triton cytoskeletons) activated Rap1 upon stretch. Rap1 guanine nucleotide exchange factor, C3G, was required for this activation; C3G as well as the adaptor protein, CrkII, in cell extract bound to Triton cytoskeletons in a stretch-dependent manner. CrkII binding, which was Cas dependent, correlated with stretch-dependent tyrosine phosphorylation of proteins in Triton cytoskeletons including Cas at the contacts with ECM. These in vitro findings were compatible with in vivo observations of stretch-enhanced phosphotyrosine signals, accumulation of CrkII at cell-ECM contacts, and CrkII Cas colocalization. We suggest that mechanical force on Triton cytoskeletons activates local tyrosine phosphorylation, which provides docking sites for cytosolic proteins, and initiates signaling to activate Rap1. PMID- 15525533 TI - Follistatin regulates enamel patterning in mouse incisors by asymmetrically inhibiting BMP signaling and ameloblast differentiation. AB - Rodent incisors are covered by enamel only on their labial side. This asymmetric distribution of enamel is instrumental to making the cutting edge sharp. Enamel matrix is secreted by ameloblasts derived from dental epithelium. Here we show that overexpression of follistatin in the dental epithelium inhibits ameloblast differentiation in transgenic mouse incisors, whereas in follistatin knockout mice, ameloblasts differentiate ectopically on the lingual enamel-free surface. Consistent with this, in wild-type mice, follistatin was continuously expressed in the lingual dental epithelium but downregulated in the labial epithelium. Experiments on cultured tooth explants indicated that follistatin inhibits the ameloblast-inducing activity of BMP4 from the underlying mesenchymal odontoblasts and that follistatin expression is induced by activin from the surrounding dental follicle. Hence, ameloblast differentiation is regulated by antagonistic actions of BMP4 and activin A from two mesenchymal cell layers flanking the dental epithelium, and asymmetrically expressed follistatin regulates the labial-lingual patterning of enamel formation. PMID- 15525534 TI - gamma-secretase functions through Notch signaling to maintain skin appendages but is not required for their patterning or initial morphogenesis. AB - The role of Notch signaling during skin development was analyzed using Msx2-Cre to create mosaic loss-of-function alleles with precise temporal and spatial resolution. We find that gamma-secretase is not involved in skin patterning or cell fate acquisition within the hair follicle. In its absence, however, inner root sheath cells fail to maintain their fates and by the end of the first growth phase, the epidermal differentiation program is activated in outer root sheath cells. This results in complete conversion of hair follicles to epidermal cysts that bears a striking resemblance to Nevus Comedonicus. Sebaceous glands also fail to form in gamma-secretase-deficient mice. Importantly, mice with compound loss of Notch genes in their skin phenocopy loss of gamma-secretase in all three lineages, demonstrating that Notch proteolysis accounts for the major signaling function of this enzyme in this organ and that both autonomous and nonautonomous Notch-dependent signals are involved. PMID- 15525535 TI - A signaling network reciprocally regulates genes associated with acute infection and chronic persistence in Pseudomonas aeruginosa. AB - The opportunistic pathogen Pseudomonas aeruginosa causes a variety of acute and chronic infections. We identified a gene whose inactivation results in attenuation of virulence due to premature activation of genes involved in biofilm formation and coordinate repression of genes required for initial colonization. This gene, retS, encodes a hybrid sensor kinase/response regulator with an unconventional arrangement of functional domains. Genome-wide transcriptional profiling indicates that the retS gene is required for expression of the Type III secretion system and other virulence factors and for repression of genes responsible for exopolysaccharide components of the P. aeruginosa biofilm matrix. These disparate phenotypes are suppressed by transposon insertions in genes encoding the GacS/GacA/rsmZ signal transduction pathway, a highly conserved system involved in the control of diverse adaptive functions. This study defines RetS as a pleiotropic regulator of multiple virulence phenotypes that orchestrates genes required for acute infection and genes associated with chronic persistence. PMID- 15525536 TI - The S. pombe Cdc14-like phosphatase Clp1p regulates chromosome biorientation and interacts with Aurora kinase. AB - The S. pombe Cdc14-related phosphatase Clp1p/Flp1p regulates G2/M transition by antagonizing CDK activity and is essential for coordinating the nuclear division cycle with cytokinesis through the cytokinesis checkpoint. At the G2/M transition, Clp1p/Flp1p is released from the nucleolus and SPB and distributes throughout the nucleus to the spindle and the contractile ring. This early relocalization is analogous to vertebrate Cdc14 homologs and stands in contrast to S. cerevisiae Cdc14p, which is not released from the nucleolus until metaphase/anaphase transition. Here, we report that Clp1p/Flp1p localizes to kinetochores in prometaphase and functions in chromosome segregation, since deletion of clp1/flp1 causes cosegregation of sister chromatids, when sister kinetochores are prone to mono-orientation. Genetic, cytological, and biochemical experiments suggest that Clp1p/Flp1p functions together with Aurora kinase at kinetochores. Together, these results suggest that Clp1p/Flp1p has a role in repairing mono-orientation of sister kinetochores. PMID- 15525538 TI - ACAP1 promotes endocytic recycling by recognizing recycling sorting signals. AB - Cargo sorting that promotes the transport of cargo proteins from a membrane compartment has been predicted to be unlikely in the endocytic recycling pathways. We now show that ACAP1 binds specifically and directly to recycling cargo proteins. Reducing this interaction for TfR inhibits its recycling. Moreover, ACAP1 binds to two distinct phenylalanine-based sequences in the cytoplasmic domain of TfR that function as recycling sorting signals to promote its transport from the recycling endosome. Taken together, these findings indicate that ACAP1 promotes cargo sorting by recognizing recycling sorting signals. PMID- 15525537 TI - RGS14 is a mitotic spindle protein essential from the first division of the mammalian zygote. AB - Heterotrimeric G protein alpha subunits, RGS proteins, and GoLoco motif proteins have been recently implicated in the control of mitotic spindle dynamics in C. elegans and D. melanogaster. Here we show that "regulator of G protein signaling 14" (RGS14) is expressed by the mouse embryonic genome immediately prior to the first mitosis, where it colocalizes with the anastral mitotic apparatus of the mouse zygote. Loss of Rgs14 expression in the mouse zygote results in cytofragmentation and failure to progress to the 2-cell stage. RGS14 is found in all tissues and segregates to the nucleus in interphase and to the mitotic spindle and centrioles during mitosis. Alteration of RGS14 levels in exponentially proliferating cells leads to cell growth arrest. Our results indicate that RGS14 is one of the earliest essential product of the mammalian embryonic genome yet described and has a general role in mitosis. PMID- 15525540 TI - Cyclooxygenase and nitric oxide systems in the gut as therapeutic targets for safer anti-inflammatory drugs. AB - After a decade of intense pharmacological and drug development activity by the pharmaceutical industry, compounds derived from two key strategies for reducing gastrointestinal effects of non-steroidal anti-inflammatory drugs (NSAIDs) have been subjected to rigorous clinical appraisal. Despite the undoubted therapeutic and commercial success of the selective cyclooxygenase (COX)-2 inhibitors, known as the coxibs, with second-generation compounds already approved and launched, some concerns over their full gastrointestinal profile still linger, while the cardiovascular safety of this class has become a key issue. Likewise, Phase II evaluation of compounds incorporating a nitric oxide (NO)-donating moiety into standard NSAIDs (the NO-NSAIDs or CINODs) has created recent controversy over the full clinical profile of these compounds. Other approaches such as NO-COX-2 inhibitors and dual COX-lipoxygenase inhibitors are already warranting interest. It might therefore be too early to predict the eventual winning strategy for safer anti-inflammatory drugs. PMID- 15525541 TI - Matrix metalloproteinases and the gut - new roles for old enzymes. AB - Matrix metalloproteinases (MMPs) are a family of neutral proteases with the ability to degrade all components of extracellular matrix. To date, more than 24 different human MMPs have been identified. MMP activity is important in diseases such as arthritis, atherosclerosis, periodontal diseases and cancer. Recent data suggest that MMPs are involved in tissue injury and healing in the human gut. PMID- 15525542 TI - Protease-activated receptors: protease signaling in the gastrointestinal tract. AB - Serine proteases from the circulation, inflammatory cells, digestive glands and microorganisms can signal to cells by cleaving protease-activated receptors (PARs), a family of four G-protein-coupled receptors. Proteases cleave PARs at specific sites to expose tethered ligand domains that bind to and activate the cleaved receptors. Despite this irreversible mechanism of activation, PAR signaling is tightly regulated to prevent the uncontrolled stimulation of cells. Although PARs are found in all organ systems, protease signaling is of particular interest in the gastrointestinal tract, where proteases regulate neurotransmission, secretion, motility, epithelial permeability and intestinal inflammation, and can thus contribute to disease. PMID- 15525543 TI - Gut endocrine secretions and their relevance to satiety. AB - The three major regions of the gastrointestinal tract (stomach, upper small intestine, ileum/colon) are each the source of factors influencing food intake. These can enhance appetite or, conversely, inhibit it. The active factors include hormones, lipid mediators, nutrients and gastric distension. Vagal afferent neurons mediate the effects of some gut signals on the brain, but other gut hormones also act directly on the brain in areas where the blood-brain barrier can be penetrated. Many different interactions between these factors are now emerging. Collectively, this system presents an array of new potential targets in seeking to modify energy intake. PMID- 15525544 TI - Gastric leptin: a new manager of gastrointestinal function. AB - Leptin, a 16 kDa protein-encoded by the ob gene, is involved in the regulation of food intake, body composition and energy expenditure through a central feedback mechanism. Initially thought to be adipocyte-specific, the ob gene, as well as the leptin receptor, has been found in a variety of other tissues including the stomach. Stomach-derived leptin, mainly secreted in the lumen, remains stable in gastric juice even at pH2. It then enters the intestine where leptin receptors have been identified on the brush border. Recent data also suggest that gut leptin may act locally within the gastrointestinal tract to influence intestinal functions, such as nutrient absorption, and thus have physiopathological implications. PMID- 15525545 TI - Growth factors and trefoil peptides in gastrointestinal health and disease. AB - Peptide growth factors are a fascinating group of molecules with diverse effects. Recent developments have allowed us to gain a much greater insight into their pathophysiological functions. In addition, the development of recombinant peptide technology, monoclonal antibody production and both artificial small molecule receptor agonists and inhibitors now allows us to use these factors for the treatment of multiple conditions including gastrointestinal malignancy (particularly colonic carcinoma), short bowel syndrome (where factors such as growth hormone, epidermal growth factor and glucagons-like peptide 2 show particular promise), and inflammatory bowel disease. PMID- 15525546 TI - The gastrointestinal pharmacology of cannabinoids: an update. AB - Recent work in the field of gastrointestinal pharmacology of cannabinoids has focused on enteric endocannabinoid and endovanilloid systems and their modulation in pathophysiological conditions. CB(1) receptor immunoreactivity was detected on enteric cholinergic neurones and vasoactive intestinal peptide-containing submucosal ganglion cells, on discrete nuclei of the dorsovagal complex (involved in emesis) and on central and peripheral vagal terminals, thus controlling gastroesophageal reflux and gastrointestinal motility. CB(1) receptor activation by endocannabinoids inhibited induced fluid secretion and inflammation in animal models and reduced proliferation of cultured colorectal cancer cells. Endocannabinoids also activate cannabinoid CB(2) and vanilloid VR1 receptors in certain inflammatory states. Thus endocannabinoid metabolism could provide a useful therapeutic target for many gastrointestinal disorders. PMID- 15525548 TI - Peptide YY and appetite control. AB - Peptide YY (PYY) is an important gut hormone synthesized and secreted by the gastrointestinal tract. Peripheral administration of PYY(3-36), one of the circulating forms of PYY, is known to inhibit food intake. This anorexigenic effect is masked by stress inhibition of appetite, and it is therefore important for animals to be thoroughly acclimatised for PYY(3-36) to be effective. Evidence suggests that PYY(3-36) acts via the hypothalamic Y(2) receptor. Levels of the anorexigenic hormone PYY(3-36) are low in overweight volunteers and could provide an important therapeutic avenue in the quest to combat the obesity epidemic. PMID- 15525549 TI - Glucagon-like peptide 1 and inhibitors of dipeptidyl peptidase IV in the treatment of type 2 diabetes mellitus. AB - Proof-of-concept for the efficacy of a glucagon-like peptide 1 (GLP-1)-based therapy of patients with type 2 diabetes was provided in 2002 by means of prolonged continuous subcutaneous infusion of native GLP-1. Since then, several long-acting analogues of GLP-1, as well as inhibitors of dipeptidyl peptidase IV, the enzyme that rapidly inactivates endogenous GLP-1, have demonstrated efficacy in long term clinical trials. PMID- 15525550 TI - 11beta-hydroxysteroid dehydrogenases: changing glucocorticoid action. AB - 11beta-hydroxysteroid dehydrogenases (11beta-HSDs) catalyse the interconversion of active cortisol and inert cortisone. Two isozymes have been discovered, each with unique properties and powerful biological roles. 11beta-HSD2 potently inactivates cortisol, protecting key tissues. By contrast, 11beta-HSD1 regenerates cortisol, amplifying its actions in liver, fat and brain. Overexpression of this isozyme may contribute to the pathogenesis of the metabolic syndrome. Its inhibition is a potential therapeutic target for both metabolic and glucocorticoid-associated CNS disorders. PMID- 15525551 TI - Mitochondrial uncoupling proteins as potential targets for pharmacological agents. AB - It is widely acknowledged that the function of the original uncoupling protein, UCP1, is uncoupling of substrate oxidation from ATP synthesis, and that its physiological purpose is thermogenesis. The mechanisms and physiological functions of the novel uncoupling proteins, identified within the past seven years, are as yet poorly understood. These novel uncoupling proteins are part of a large family comprising approximately 35 mitochondrial anion carrier proteins. UCP2 and UCP3 appear to function in reactive oxygen species handling and/or in fatty acid metabolism; uncoupling might occur secondarily. There is little information on UCP4 and UCP5 (BMCP1), and phylogenetic analyses indicate that they are further removed from UCP1 than mitochondrial anion carrier proteins, and have distinct functions. PMID- 15525552 TI - Somatostatin analogs - from new molecules to new applications. AB - Somatostatin (SST) was firstly discovered as a hypothalamic hormone inhibiting GH secretion. Despite its broad inhibitory effects on both endocrine and exocrine secretions, natural SST has limited therapeutic potential owing to its short plasma half-life. The synthesis of the first two metabolically stabilized and more potent SST analogs (octreotide and lanreotide) established the use of SST peptide therapy. The discovery of the five SST receptor (sst(1-5)) subtypes in the 1990s further enhanced our understanding of the biological roles of SST, created new therapeutic opportunities and highlighted the limitation of 'classical' SST analogs, which act mainly via receptor subtype 2 and are unable to reproduce all actions of native SST. To diminish these limitations, new SST analogs highly selective for particular receptor subtypes, together with so called 'universal' analogs acting on multiple receptor subtypes, have been developed. These compounds have shown promise in preclinical studies and might further advance the use of SST analog therapy in the future. The development of SST analogs coupled to radioisotopes or cytotoxic drugs, which allows the selective destruction of tumor cells overexpressing sst receptors, constitutes another field of progress. PMID- 15525553 TI - Abuse of anabolic androgenic steroids and related substances in sport and exercise. AB - Anabolic androgenic steroids are synthetic derivatives of testosterone, which is the primary male sex hormone. Anabolic androgenic steroids are used to enhance athletic performance and appearance. Adverse effects include those on the liver, serum lipids, psyche/behavior and reproductive system. Androstenedione is an anabolic androgenic steroid used to increase blood testosterone levels for the purposes of increasing strength, lean body mass and sexual performance. However, there is no research indicating that androstenedione, or its related compounds, significantly increases strength and/or lean body mass in humans by increasing testosterone levels. The long-term health effects of prolonged androstenedione supplementation are unknown. Dehydroepiandrosterone (DHEA) is a weak androgen also used to elevate testosterone levels, and is advertised as an anti-obesity and anti-aging supplement capable of improving libido, vitality and immunity levels. However, research demonstrates that DHEA supplementation does not increase serum testosterone concentrations or increase strength in men, and may acutely increase testosterone levels in women, thus producing a virilizing effect. PMID- 15525554 TI - Hormonal and metabolic strategies to attenuate catabolism in critically ill patients. AB - During the prolonged phase of critical illness, the ongoing hypermetabolic response leads to loss of lean tissue mass. Although the cachexia of prolonged illness is usually associated with low concentrations of anabolic hormones, most endocrine interventions attempting to correct the hormone balance have shown to be ineffective and their indiscriminate use is even harmful. Thus, a detailed understanding of the neuroendocrinology of the stress response is warranted, especially as the acute and chronic phases show remarkable differences. In the acute stress response, low circulating peripheral anabolic hormone levels, despite an actively secreting pituitary, are indicative of peripheral resistance to the anterior pituitary hormones. By contrast, the pulsatile secretion of anterior pituitary hormones is uniformly decreased in the prolonged phase of the disease, leading to proportionally reduced concentrations of peripheral anabolic hormones. As hypothalamic secretagogues can restore the pulsatile secretion of the anterior pituitary and increase peripheral target hormones, tissues are at least partially sensitive to the anterior pituitary hormones in this phase of illness. Therefore, a combination of hypothalamic secretagogues that reactivates the anterior pituitary to a greater extent could be a more physiological and effective strategy to induce anabolism in patients with prolonged critical illness. PMID- 15525555 TI - The molecular complexity of glucocorticoid actions in inflammation - a four-ring circus. AB - Glucocorticoids have been the mainstay of anti-inflammatory therapy for the past 50 years despite the significant risks associated with their long-term use. Recent research into the molecular mechanisms of action of these compounds can be aggregated into two major themes: first, those that focus on the involvement of the glucocorticoid receptor in regulating the transcription of pro-inflammatory genes; and second, studies that seek to identify alternative pathways of glucocorticoid action either via the induction of anti-inflammatory mediators or through rapid non-genomic mechanisms. Understanding the temporal-spatial patterns of these multiple mechanisms on relevant cell types will be key to explaining the profound effects that these four-ring steroids exert on the inflammatory process. PMID- 15525556 TI - Disseminated Mycobacterium haemophilum infection as initial manifestation of AIDS. AB - Mycobacterium haemophilum has rarely been implicated in human disease. The organisms have been isolated mainly in patients with human immunodeficiency virus (HIV) disease or transplant recipients. We describe the first case of a disseminated M. haemophilum infection as initial manifestation of AIDS in Europe. PMID- 15525557 TI - The toll-like receptor 4 Asp299Gly variant: no influence on LPS responsiveness or susceptibility to pulmonary tuberculosis in The Gambia. AB - SETTING: Tuberculosis (TB) remains a major cause of morbidity and mortality worldwide. Studies in a murine model of pulmonary TB have identified a role for Toll-like receptor 4 (TLR4) in the development of chronic lung infection with Mycobacterium tuberculosis. The Asp299Gly polymorphism in the human TLR4 gene is associated with in vivo hyporesponsiveness to lipopolysaccharide (LPS) in Caucasians. OBJECTIVE: To determine whether TLR4 Asp299Gly influences LPS responses or susceptibility to pulmonary TB in humans in a Gambian population sample. DESIGN: We compared whole blood monokine responses to LPS in 245 healthy blood donors stratified by TLR4 Asp299Gly genotype to assess whether this polymorphism was functional in this population. A case-control study of 640 subjects was conducted to investigate whether TLR4 Asp299Gly was associated with TB. RESULTS: LPS-induced tumour necrosis factor, interleukin-1 beta and interleukin-10 production was not influenced by TLR4 Asp299Gly genotype. There was no association between TLR4 Asp299Gly and TB. CONCLUSION: Our data suggest that TLR4 Asp299Gly has no influence on monocyte LPS responses or susceptibility to TB in Gambians and could be an ancient neutral polymorphism. PMID- 15525558 TI - Development of a scintillation proximity assay for the Mycobacterium tuberculosis KasA and KasB enzymes involved in mycolic acid biosynthesis. AB - Tuberculosis remains a global health problem, and programs dedicated to discovery of novel compounds against Mycobacterium tuberculosis require robust assays for high-throughput screening of chemical and natural product libraries. Enzymes involved in the biosynthesis of mycolic acids, vital components of the mycobacterial cell wall, have received much attention as potential drug targets. KasA and KasB, examples of the beta-ketoacyl-acyl carrier protein synthase I/II (KASI/II) class of condensing enzymes of the M. tuberculosis fatty acid synthase II system have been the focus of several studies designed to biochemically characterize these enzymes. Whilst robust methods have been developed for FabH like proteins, fast and sensitive assays for high-throughput screening of KASI/II enzymes have not been available. Here we report the development of a direct scintillation proximity assay (SPA) for the KASI/II enzymes, KasA and KasB. The SPA was more sensitive than existing assays, as shown by its ability to measure activity using less enzyme than other assay formats, and the SPA was validated using the known KAS inhibitor thiolactomycin. In addition, the KasA and KasB SPA was adapted for use with Staphylococcus aureus FabF to show the versatility of this assay format to KAS enzymes from other pathogenic organisms. PMID- 15525559 TI - Statistical limitations to the Cornell model of latent tuberculosis infection for the study of relapse rates. AB - The Cornell model has been extensively used as a mouse model for studying the latent stage of Mycobacterium tuberculosis infection. In this model mice are infected and then given a course of chemotherapy prior to reactivation of infection. We discuss here the importance of using adequate mouse numbers in a Cornell model for the study of relapse rates in order to obtain sufficient statistical power to confirm a hypothesis. Experiments with small sample sizes are useful for 'screening' experiments, but will have very little value in 'confirming' the objective. When the objective of the experiment is confirmation of an effect through establishment of statistical significance, power calculations are critical in order to assure that the sample size will be sufficient to meet that objective. PMID- 15525560 TI - Pharmacokinetics and relative bioavailability of clofazimine in relation to food, orange juice and antacid. AB - BACKGROUND: Clofazimine is potentially useful for the treatment of disease due to multidrug resistant Mycobacterium tuberculosis, as well as leprosy and certain chronic skin diseases. Its pharmacokinetics have been incompletely characterized. This study was conducted to explore issues relating to bioavailability in the presence of food, orange juice, and antacid. METHODS: A 5 drug regimen consisting of clofazimine, cycloserine, ethionamide, para-aminosalicyclic acid, and pyridoxime was administered to healthy subjects four times using a four period cross-over design with two weeks washout between treatments. Subjects also received orange juice, a high fat meal, aluminum/magnesium antacid, or only water in random order with the drug regimen. The pharmacokinetics of clofazimine were assessed using individual- and population-based methods and relative bioavailability compared to fasting administration was determined. RESULTS: Clofazimine exhibited a sometimes prolonged and variable lag-time and considerable variability in plasma concentrations. From the population analysis (one-compartment model), the mean oral clearance was 76.7 l/h (CV=74.2%) and mean apparent volume of distribution was 1470 l (CV=36.3%). The first-order absorption rate constant ranged from 0.716 to 1.33 h(-1) (pooled CV=61.7%). Residual (proportional) error was 49.1%. Estimates of bioavailability compared to fasting administration were 145% (90% CI, 107-183%) for administration with high fat food, 82.0% (63.2-101%) for administration with orange juice, and 78.5% (55.1 102%) for administration with antacid. CONCLUSION: Administration of clofazimine with a high fat meal provides the greatest bioavailability, however, bioavailability is associated with high inter- and intra-subject variability. Both orange juice and aluminum-magnesium antacid produced a reduction in mean bioavailability of clofazimine. PMID- 15525561 TI - Morphometric analysis of Th(1) and Th(2) cytokine expression in human pulmonary tuberculosis. AB - SETTING: Following infection with Mycobacterium tuberculosis, host cytokine responses influence disease manifestation. Differences in cytokine expression likely determine whether tuberculosis (TB) progresses, resolves, or becomes latent. In particular, the balance between Th(1) and Th(2) cytokine responses influences the expression of disease in individuals with pulmonary TB. OBJECTIVE AND DESIGN: Since the cytokine microenvironment in pulmonary TB remains suboptimally defined, we utilized quantitative immunohistochemistry to compare the expression of Th(1) cytokines [interferon-gamma (IFNgamma) and interleukin-12 (IL-12)] and Th(2) cytokines [IL-4, IL-10, transforming growth factor-beta (TGFbeta)] in surgically resected lungs of seven TB patients and four control subjects. We also quantified IFNgamma-inducible protein 10 (IP-10) expression, a CXC chemokine for macrophages and T cells. RESULTS: Morphometric analyses revealed increased IFNgamma, IL-12, IP-10, and TGFbeta in granulomas and in pneumonitis areas of TB lungs. In contrast, IL-10 and IL-4 expressions were globally reduced in TB lung tissues compared to controls. CONCLUSION: Th(1) cytokines and TGFbeta are increased while Th(2) cytokines are decreased in well formed pulmonary granulomas of TB patients compared to controls. PMID- 15525562 TI - A matrix metalloproteinase inhibitor promotes granuloma formation during the early phase of Mycobacterium tuberculosis pulmonary infection. AB - OBJECTIVE: The host response to pulmonary Mycobacterium tuberculosis (Mtb) infection results in granuloma formation in an effort to limit infection, but the host immune cells also provide an environment in which Mtb persists. Granuloma formation requires immune cell infiltration and concurrent extensive remodeling of pulmonary tissue which we hypothesize to be the result of increased matrix metalloproteinases (MMP) activity. DESIGN: C57BL/6 mice infected with virulent Mtb (H37Rv) via intratracheal inoculation were treated with a synthetic inhibitor of MMP activity (BB-94). Mice were assessed for colony forming units, granuloma morphology, leukocyte recruitment and cytokine levels over 90 days of infection. RESULTS: BB-94 treated mice had significantly decreased numbers of pulmonary and blood-borne Mtb early during disease, increased collagen deposition within early granulomas and significantly decreased pulmonary leukocyte recruitment when compared to vehicle-treated, Mtb-infected mice. Cytokine expression did not differ significantly between groups. CONCLUSION: Events of early granuloma formation can be modified by inhibiting MMP activity, by decreasing leukocyte recruitment, a major source of MMPs during infection, enhancing the establishment of granulomas and decreasing blood-borne dissemination of Mtb. PMID- 15525563 TI - Short- and long-term spatial delayed response performance across the lifespan. AB - Delayed response paradigms have been used to examine the neural basis of short and long-term memory in humans. However, limited information exists on how delayed response performance changes across the lifespan. Using a well-validated spatial delayed response (SDR) task, we examined performance at short and long delays in over 300 control participants, 7 to 80 years old. We found a significant nonlinear relation between age and short delay performance (children and older adults worse than young adults) and a significant effect of delay length across the entire lifespan (long worse than short; largest in the youngest children, diminishing nonlinearly with age). This study compares short and long term spatial memory and suggests that the relation between these systems may alter across the lifespan. PMID- 15525565 TI - Age-associated memory changes in adults with williams syndrome. AB - Age-associated changes on measures of episodic and working memory were examined in 15 adults with Williams Syndrome (WS; M age = 48.3 years, SD = 14.7; M IQ = 62.9, SD = 8.5) and their performance was compared to that of 33 adults with mental retardation (MR) with unspecified etiologies (M age = 54.2 years, SD = 8.9; M IQ = 61.7, SD = 6.5). Among the group with WS, older adults were significantly poorer than younger adults on the free recall task, a measure of episodic memory. Although this finding is consistent with normal aging, it occurred at a chronologically early age in adults with WS and was not found in their peers with unspecified MR. Although both groups showed small declines with age on a backward digit span task, a measure of working memory, for the group with WS the rate of decline on backward digit span was slower as compared to their performance on the free recall task. The findings from this study indicate a chronologically early and precipitous age-associated decrease in long-term, episodic memory in adults with WS. PMID- 15525564 TI - A retrospective neurocognitive study in children with spastic diplegia. AB - The study presents the results on neonatal cranial ultrasonography (US) and later intelligence (Wechsler Intelligence Scale-Third Edition and Wechsler Preschool and Primary Scale of Intelligence-Revised) and Neuropsychological assessments of 15 children with spastic diplegia. The assessments were undertaken when the children were 5 to 12 years of age. The children's IQ scores were, as a group, at the lower end of the normal distribution. The neuropsychological assessment indicated that deficits in visuomotor and visuospatial processing were characteristic of the children. No association was found between the neonatal cranial US findings and the IQ and neurocognitive scores. However, the cranial US findings strongly predicted functional motor limitations of the children. PMID- 15525566 TI - Social competence in young children with inflicted traumatic brain injury. AB - As infants develop skills that allow for increasing independence in social and cognitive domains, they acquire the ability to identify goals, sequence behaviors to carry out goals, and to flexibly use strategies for attaining goals in both social and independent play contexts. Little is known about how brain injury in young children may disrupt the precursors to such executive processes. In this study, we examined social and cognitive competence in 25 infants ages 3 to 23 months who sustained moderate to severe traumatic brain injury (TBI) secondary to physical abuse and in 22 healthy community comparison children. Children with TBI were evaluated an average of 1.6 months after the injury. A toy-centered activity with the examiner was used to capture joint attention and social behavior and an exploratory toy play situation was used to measure independent goal-directed play. The inflicted TBI group showed significant reduction in both social and cognitive domains relative to the comparison group. Canonical correlation analyses disclosed that inflicted TBI was associated with reduction in (a) initiation of social interactions, (b) responsiveness to interactions initiated by the examiner, (c) positive affect, and (d) compliance. The groups performed comparably on indexes of gestural and verbal communication and for the occurrence of negative affect. Joint attention was an area of vulnerability for the TBI group in both social initiation and response contexts. Although general cognitive and motor scores were lower in the inflicted TBI group, the complexity of independent toy play did not differ across groups. Early brain injury causes significant disruption in behaviors regulating initiation and responsiveness in social contexts. Longitudinal follow-up will characterize the long-term consequences of early disruption in joint attention and other behaviors on the development of social and cognitive precursors to executive processes. PMID- 15525567 TI - Utilization behavior in boys with ADHD: a test of Barkley's theory. AB - Barkley's (1997) comprehensive theory of attention deficit hyperactivity disorder (ADHD) predicts that affected children express frontal lobe related impairments, including expression of utilization behavior-the appropriate use of an object in an inappropriate context. Nineteen boys with and 20 without ADHD were exposed to a series of utilitarian (potentially utilization-inducing) and neutral objects. Boys in the ADHD group exhibited more utilization behavior and did so more quickly than boys in the control group; engagement with utilitarian objects and tendencies to do so quickly best predicted ADHD and control group membership. Beyond utilization behavior distinctions, general impulsivity differences were seen between boys with ADHD and control group counterparts, but these were less pronounced than the utilization behavior differences. Results offer support for utilization behavior's place in Barkley's theory. PMID- 15525568 TI - The influence of unequal numbers of trials on comparisons of average event related potentials. AB - Four studies examined (a) how event-related potentials (ERPs) change as the number of trials averaged increases and (b) the statistical implications of comparing ERPs composed of different numbers of trials. Experiment 1 utilized data from 7-year-old children performing an oddball task. The other three experiments used simulated data with different distributions of P3 peak latency. In all 4 experiments, peak amplitude decreased and the mean amplitude of the 300 to 900 msec interval remained stable as the number of trials averaged increased. The standard deviations of both measures decreased. These data show that the decrease in peak amplitude with increasing numbers of trials that has been found in other studies is not solely due to the elimination of residual noise but is likely to also involve a fundamental aspect of signal averaging and the algorithm used to select peaks. Furthermore, these experiments expose the possibility of statistical errors when investigators compare average ERPs composed of small versus large numbers of trials as is often done when the oddball paradigm is used. PMID- 15525569 TI - Effects of GnRH antagonist treatment on follicular development and angiogenesis in the primate ovary. AB - Angiogenesis is required for normal follicular development but the role of gonadotrophins in the control of follicular angiogenesis remains to be elucidated. This study investigated the effects of treatment with GnRH antagonist in vivo on follicular development and angiogenesis in the marmoset. GnRH antagonist was administered on either follicular day 0 or day 5 of the 10-day follicular phase with ovaries collected on day 10. Ovaries from control marmosets were studied at day 5 (mid follicular phase) and day 10 (periovulatory period). Ovaries were fixed, serial sectioned and subjected to morphological analysis and immunocytochemistry to determine cell proliferation and follicular endothelial cell area and in situ hybridization to assess changes in expression of vascular endothelial growth factor (VEGF). Treatment with GnRH antagonist from day 0-10 resulted in an absence of dominant preovulatory follicles seen in controls. In the remaining tertiary follicles granulosa, theca and endothelial cell proliferation was reduced, resulting in a minor reduction in vascular density. However, VEGF mRNA expression was unaffected by treatment. Treatment from day 5 10 did not prevent development of ovulatory size follicles, but they were atretic and lacked VEGF mRNA. These results suggest that while VEGF expression in the preovulatory follicle is under gonadotrophic control it is not dependent on normal gonadotrophin secretion in tertiary follicles, indicating that there are other paracrine factors regulating VEGF expression in the developing ovarian follicle. PMID- 15525570 TI - Steroid signalling in human ovarian surface epithelial cells: the response to interleukin-1alpha determined by microarray analysis. AB - The human ovarian surface epithelium (HOSE) is a common site of gynaecological disease including endometriosis and ovarian cancer, probably due to serial injury repair events associated with successive ovulations. To comprehend the importance of steroid signalling in the regulation of the HOSE, we used a custom microarray to catalogue the expression of over 250 genes involved in the synthesis and reception of steroid hormones, sterols and retinoids. The array included a subset of non-steroidogenic genes commonly involved in pro-/anti-inflammatory signalling. HOSE cells donated by five patients undergoing surgery for non malignant gynaecological conditions were cultured for 48 h in the presence and absence of 500 pg/ml interleukin-1alpha (IL-1alpha). Total RNA was reverse transcribed into biotin-labelled cDNA, which was hybridised to the array and visualised by gold-particle resonance light scattering and charge-coupled device (CCD) camera detection. Results for selected genes were verified by quantitative reverse-transcription PCR. In five out of five cases, untreated HOSE cells expressed genes encoding enzymes required for de novo biosynthesis of cholesterol from acetate and subsequent formation of C21-pregnane and C19-androstane steroids. Consistent with the inability of HOSE cells to synthesise glucocorticoids, oestrogens or 5alpha-reduced androgens de novo, CYP21, CYP19 and 5alpha-reductase were not detected. The only steroidogenic gene significantly up regulated by IL-1alpha was 11beta-hydroxysteroid dehydrogenase type 1 (11betaHSD1). Other cytokine-induced genes were IL-6, IL-8, nuclear factor kappaB (NFkappaB) inhibitor alpha, metallothionein-IIA and lysyl oxidase: inflammation associated genes that respond to glucocorticoids. The only steroidogenic gene significantly suppressed by IL-1alpha was 3betaHSD1. Other genes suppressed by IL 1alpha were aldehyde dehydrogenase (ALDH) 1, ALDH 10, gonadotrophin hormone releasing hormone receptor, peroxisome proliferation-activated receptor-binding protein (PPAR-bp) and nuclear receptor subfamily 2 group F member 2. These results define a steroidogenic phenotype of cultured HOSE cells and provide a limited expression profile for genes with associated signalling functions. IL 1alpha co-ordinately induces 11betaHSD1 and a panel of glucocorticoid-regulated, inflammation-associated genes in HOSE cells, providing further evidence that cortisol generated by 11betaHSD1 could participate in the local resolution of inflammation associated with ovulation. PMID- 15525571 TI - Reproductive failure in mice lacking inter-alpha-trypsin inhibitor (ITI)--ITI target genes in mouse ovary identified by microarray analysis. AB - Bikunin, a Kunitz-type protease inhibitor, is found in blood and urine. It has been established by two laboratories independently that the bikunin knockout female mice display a severe reduction in fertility: the cumulus oophorus has a defect in forming the extracellular hyaluronan-rich matrix during expansion. Proteins of the inter-alpha-trypsin inhibitor (ITI) family are eliminated in mice in which the bikunin gene has been inactivated, since bikunin is essential for their biosynthesis. Proteins of the ITI family may contribute to the microenvironment in which ovulation takes place. It is not clear, however, whether a single mechanism affects the reproductive function including ovulation. For identifying the full repertoire of the ITI deficiency-related genes, a cDNA microarray hybridization screening was conducted using mRNA from ovaries of wild type or bik(-/-) female mice. A number of genes were identified and their regulation was confirmed by real-time RT-PCR analysis. Our screen identified that 29 (0.7%) and 5 genes (0.1%) of the genes assayed were, respectively, up- and down-regulated twofold or more. The identified genes can be classified into distinct subsets. These include stress-related, apoptosis-related, proteases, signaling molecules, aging-related, cytokines, hyaluronan metabolism and signaling, reactive oxygen species-related, and retinoid metabolism, which have previously been implicated in enhancing follicle development and/or ovulation. Real-time RT-PCR analysis confirmed that these genes were up- and down-regulated two- to tenfold by bikunin knockout. These studies demonstrate that proteins of the ITI family may exert potent regulatory effects on a major physiological reproductive process, ovulation. PMID- 15525572 TI - Changes in cAMP-dependent protein kinase (PKA) and progesterone secretion in luteinizing human granulosa cells. AB - Luteinization of follicular granulosa cells leads to an increase in progesterone secretion that is regulated by luteinizing hormone (LH). LH acts mainly by elevating intracellular cyclic 3',5'-adenosine monophosphate (cAMP) and activating cAMP-dependent protein kinase (PKA). In this study, we have examined the role of PKA in relation to progesterone output by luteinizing human granulosa cells. Human granulosa cells were obtained by percoll gradient centrifugation of follicular aspirates of patients undergoing oocyte retrieval for assisted conception. Cells were cultured in serum-supplemented medium for up to 3 days in the presence and/or absence of human (h)LH and other cAMP-elevating agents. Spent medium was assayed for cAMP and progesterone content by specific RIA. Cell lysates were collected and assessed for PKA regulatory (R)IIalpha/catalytic (C)alpha expression by Western blotting. Although basal progesterone secretion increased progressively throughout culture, cAMP levels remained unchanged. Under basal conditions, PKA RIIalpha/Calpha expression appeared to increase throughout the 3-day culture period. In the presence of hLH and other cAMP-elevating agents, progesterone secretion increased in a dose-dependent manner coincident with an increase in cAMP. However, despite the increase in both progesterone secretion and cAMP accumulation, there was a dose-dependent decrease in both PKA RIIalpha and Calpha expression. Thus, data presented in this study show that increases in progesterone secretion in luteinizing human granulosa cells can be dissociated from increases in PKA expression. This notion implies that progesterone secretion may be regulated by PKA-dependent as well as PKA-independent mechanisms. PMID- 15525573 TI - Progesterone secretion by luteinizing human granulosa cells: a possible cAMP dependent but PKA-independent mechanism involved in its regulation. AB - The corpus luteum formed after luteinization of follicular cells secretes progesterone under the control of luteinizing hormone (LH). Binding of LH to its G-protein-coupled receptor leads to the activation of the adenylate cyclase/ cyclic AMP (cAMP)/cAMP-dependent protein kinase (PKA) signalling pathway. The identification of a new class of cAMP-binding proteins termed 'guanine nucleotide exchange factors' (cAMP-GEFs) provides a means by which changes in cAMP could yield actions that are independent of PKA. Hence, in this study, we have explored the hypothesis that steroidogenesis in luteinizing cells is mediated in both a cAMP/PKA-dependent and cAMP-dependent, but PKA-independent, manner. Human granulosa cells were isolated from follicular aspirates of women undergoing assisted conception. Luteinizing human granulosa cells were cultured for up to 3 days in the presence of human (h)LH and the adenylate cyclase activator forskolin in the added presence or absence of increasing doses of the PKA inhibitors H89 (N [2-(4-bromocinnamylamino)ethyl] 5-isoquinoline) and PKI (myristoylated protein kinase A inhibitor amide 14-22) or the cAMP antagonist, Rp-cAMP. Agonist stimulated progesterone secretion was inhibited in a dose-dependent manner by the PKA inhibitors and the cAMP antagonist, with decreasing sensitivity as luteinization progressed. Pretreatment of granulosa cells for 4 h with human (h)LH reduced the effectiveness of H89 in inhibiting progesterone secretion. Under basal conditions, cAMP-GEFI expression increased progressively throughout culture, and this could be further enhanced when cells were incubated with increasing doses of LH and forskolin. Furthermore, incubation of cells in the presence of increasing concentrations of the novel cAMP-GEF-specific cAMP analogue, 8 CPT-2 ME-cAMP (8-(4-chloro-phenylthio)-2'-0-methyladenosine-3',5' cyclic monophosphate), increased progesterone secretion in a dose-dependent manner. The results show that increases in cAMP generated by LH and forskolin, in addition to activating PKA, also induce increases in cAMP-GEFI protein expression in luteinizing human granulosa cells. In addition, activation of cAMP-GEFI results in increased progesterone secretion. Hence, increases in cAMP lead to the activation of PKA-dependent, as well as PKA-independent but cAMP-dependent (via cAMP-GEFI), signalling mechanisms. Since cAMP-GEFs have the capacity to activate the mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3-K)/protein kinase B (PKB) signalling pathways, these may provide the potential mechanisms by which cAMP-dependent but PKA-independent progesterone synthesis is regulated. PMID- 15525574 TI - A switch from continuous to episodic testicular testosterone release in response to pulsatile LH stimulation in juvenile rhesus monkeys (Macaca mulatta). AB - This study examined the ontogeny of the testicular testosterone response to precocious pulsatile LH stimulation in the juvenile rhesus monkey. LH stimulation was achieved with an i.v. infusion (one pulse every 3 h) of either single-chain human (sch)LH, administered alone or in combination with recombinant human (rh)FSH, or recombinant monkey (rm)LH in combination with rmFSH. Homologous gonadotropin treatment resulted in an adult profile of circulating mLH concentrations. The schLH infusions produced a similar pulsatile pattern in circulating LH with peak concentrations of approximately 5 IU/l. Although a robust testicular testosterone response was observed after 24 h of intermittent LH stimulation, surprisingly testosterone release at this time was continuous. The apulsatile mode of testosterone secretion, however, did not persist, and a switch to an unequivocal episodic mode of secretion, comparable to that observed in adult monkeys, occurred by day 4 of LH stimulation. FSH did not influence the pattern of the testosterone response. We conclude from these findings that progenitor Leydig cells in the primate testis are able to respond rapidly to a physiological LH stimulus. While the cell biology underlying the switch from a continuous to a pulsatile mode of testosterone secretion remains unclear, we suggest that this phenomenon may be related to the hypothesis that episodic testosterone secretion is required for the operation of the neuroendocrine axis governing testicular function. PMID- 15525575 TI - Hormonal regulation of H19 gene expression in prostate epithelial cells. AB - The H19 gene is transcribed in an mRNA-like noncoding RNA. When tumors of various organs or cell types are considered, H19 oncogene or tumor-suppressor status remains controversial. To address the potential regulation of H19 gene expression by an androgen steroid hormone (DHT: dihydrotestosterone) or by a peptidic hormone (PRL: prolactin), we performed experiments in rats systemically treated with chemical mediators. This range of in vivo experiments demonstrated that chronic hyperprolactinemia upregulated the H19 expression in epithelial and stromal cells whereas DHT downregulated the gene. PRL and DHT appeared to be opposite mediators in the H19 RNA synthesis. We investigated these hormonal effects in three human prostate epithelial cell lines. In LNCaP cancer cells, the opposite effect of PRL and DHT was corroborated. However, in normal cells (PNT1A), H19 remained insensitive to the hormones in fetal calf serum (FCS) medium but became responsive in a serum-stripped medium. In the DU-145 cancer cell line, tested for its androgen-independence and aggressiveness, the hormones had no effect on H19 expression whatever the culture conditions. Finally, we demonstrated that PRL upregulated the H19 expression in LNCaP cells by the JAK2 STAT5 transduction pathway. We conclude that H19 expression is regulated by both a peptidic and a male steroid hormone. PMID- 15525576 TI - The mitochondrial-dependent pathway is chronically affected in testicular germ cell death in adult rats exposed in utero to anti-androgens. AB - In utero exposure to exogenous anti-androgenic compounds induces a wide range of abnormalities of the reproductive system, including hypospermatogenesis, cryptorchidism and hypospadias. By using rats exposed in utero to the anti androgenic compound flutamide (0.4, 2 or 10 mg/kg per day), it has been shown that hypospermatogenesis in adult testes could be related to (i) a long-term apoptosis in germ cells but not in somatic Leydig and Sertoli cells as evidenced by the TUNEL approach and (ii) alterations in the mRNA and protein expression of pro- (Bax, Bak, Bid) and anti-apoptotic (Bcl-2, Bcl-w) members of the Bcl-2 family. Indeed, the number of apoptotic germ cells increased with the dose of flutamide administered and the apoptotic germ cells were mainly detected at androgen-dependent stages VII-VIII. Moreover, for the Bcl-2-related proteins that were expressed mainly in the germ cells, a decrease in the levels of anti apoptotic peptides Bcl-w (60%, P=0.003) and Bcl-2 (90%, P=0.0001) was observed at 2 mg/kg per day flutamide and an increase in levels of the pro-apoptotic Bax (2.3 fold, P=0.0004) was detected at 10 mg/kg per day. In contrast, the levels of pro apoptotic peptide Bak that was mainly expressed in somatic cells decreased (70%, P=0.0008) at 10 mg/kg per day. Such alterations in Bcl-2-related peptides occurred mainly at the protein level except for Bcl-2 (72%, P=0.0001) and Bak (43%, P=00002) transcripts. Together, these results showed that the apoptosis observed in adult germ cells from rats exposed in utero to flutamide may result from a long-term alteration in the balance between pro- and anti-apoptotic Bcl-2 related molecules in favour of pro-apoptotic proteins. These data further supported the concept of an androgen-dependent fetal programming that is in relation with an alteration of the expression of Bcl-2-related genes/proteins promoting apoptosis in testicular germ cells of adult rats with fetal androgen disruption. PMID- 15525577 TI - The effects of steroidal estrogens in ACI rat mammary carcinogenesis: 17beta estradiol, 2-hydroxyestradiol, 4-hydroxyestradiol, 16alpha-hydroxyestradiol, and 4-hydroxyestrone. AB - Several investigators have suggested that certain hydroxylated metabolites of 17beta-estradiol (E2) are the proximate carcinogens that induce mammary carcinomas in estrogen-sensitive rodent models. The studies reported here were designed to examine the carcinogenic potential of different levels of E2 and the effects of genotoxic metabolites of E2 in an in vivo model sensitive to E2 induced mammary cancer. The potential induction of mammary tumors was determined in female ACI rats subcutaneously implanted with cholesterol pellets containing E2 (1, 2, or 3 mg), or 2-hydroxyestradiol (2-OH E2), 4-hydroxyestradiol (4-OH E2), 16alpha-hydroxyestradiol (16alpha-OH E2), or 4-hydoxyestrone (4-OH E1) (equimolar to 2 mg E2). Treatment with 1, 2, or 3 mg E2 resulted in the first appearance of a mammary tumor between 12 and 17 weeks, and a 50% incidence of mammary tumors was observed at 36, 19, and 18 weeks respectively. The final cumulative mammary tumor incidence in rats treated with 1, 2, or 3 mg E2 for 36 weeks was 50%, 73%, and 100% respectively. Treatment of rats with pellets containing 2-OH E2, 4-OH E2, 16alpha-OH E2, or 4-OH E1 did not induce any detectable mammary tumors. The serum levels of E2 in rats treated with a 1 or 3 mg E2 pellet for 12 weeks was increased 2- to 6-fold above control values (approximately 30 pg/ml). Treatment of rats with E2 enhanced the hepatic microsomal metabolism of E2 to E1, but did not influence the 2- or 4 hydroxylation of E2). In summary, we observed a dose-dependent induction of mammary tumors in female ACI rats treated continuously with E2; however, under these conditions 2-OH E2, 4-OH E2, 16alpha-OH E2, and 4-OH E1 were inactive in inducing mammary tumors. PMID- 15525578 TI - Sex steroid regulation and identification of different transcription units of the SA gene in mouse kidney. AB - Although the SA gene was first identified as a putative candidate gene to understand the molecular basis of hypertension in rat and humans, the concept has not been supported in recently generated SA-null mice. We had first identified the mouse SA gene on the basis of its strong androgenic regulation in mouse kidney and further characterized its genomic organization, transcription start site and chromosomal location. Northern blot, RT-PCR and in situ hybridization assays determined mouse strain, tissue distribution, sex-hormone dependence and cell expression of the SA) mRNA. Kidney and liver constitute the main expression sites of the SA gene; in particular it is expressed in epithelial proximal tubule cells in the presence of androgens. This androgen-dependent expression is abrogated when estrogens are also present. By using the sensitive RT-PCR technique, minor SA expression sites, corresponding to testes, stomach, heart and lung, have also appeared. Like in kidney, expression of the SA gene in heart and lung is androgen-dependent. Production of rabbit antibodies against SA-synthetic peptides identified the SA protein, a moiety of unknown function, which has been defined as a member of the acyl-CoA synthetase family. We have determined that the SA protein follows the same distribution and regulation as its corresponding mRNA. Transient transfection assays followed by confocal microscopy identified the mitochondria of proximal tubule-derived PCT3 cells as the subcellular location of the SA protein. Different transcriptional units produced by splicing events, occurring before the translation initiation site, have been identified from mouse kidney. This work provides the basis to further understand the molecular mechanisms that control the sex-steroid-dependent expression of the SA gene in mouse kidney, heart and lung, where SA is also expressed in an androgen dependent manner. PMID- 15525579 TI - Studies of the neuromedin U-2 receptor gene in human obesity: evidence for the existence of two ancestral forms of the receptor. AB - Central administration of neuromedin U (NMU) suppresses food intake acting through the NMU-2 receptor (NMU2R), which is expressed in the hypothalamus. We screened the NMU2R gene in 96 patients with severe early-onset obesity. A common variant haplotype was found (f-0.21). This common variant haplotype was unusual in nature, consisting of four non-contiguous missense changes in complete linkage disequilibrium, and across two separate exons. The variant haplotype resulted in four amino acid substitutions (S295T/F312L/P380L/ M385 V) and was present in several other Europid populations and in subjects of South Asian, East Asian and African American origin, but not in eleven African Pygmies. This variant haplotype was not associated with obesity or related traits in 500 subjects from a prospective population-based cohort. In summary, we have identified two markedly different isoforms of the NMU-2 receptor, presumably arising through an ancient and complex mutational event; no genetic associations between this haplotype and obesity-related traits were, however, discerned. Further investigation of the pharmacogenomic consequences of NMU2R variation in humans is warranted. PMID- 15525580 TI - Influence of genotype on the differential ontogeny of uncoupling protein 2 and 3 in subcutaneous adipose tissue and muscle in neonatal pigs. AB - The present study aimed to determine whether porcine genotype and/or postnatal age influenced mRNA abundance or protein expression of uncoupling protein (UCP)2 or 3 in subcutaneous adipose tissue (AT) and skeletal muscle (SM) and the extent to which these differences are associated with breed-specific discordance in endocrine and metabolic profiles. Piglets from commercial and Meishan litters were ranked according to birth weight. Tissue samples were obtained from the three median piglets from each litter on either day 0, 4, 7, 14 or 21 of neonatal life. UCP2 protein abundance in AT was similar between genotypes on the first day of life, but it was elevated at all subsequent postnatal ages (P<0.05) in AT of Meishan piglets. In contrast, UCP2 mRNA abundance was lower in Meishans up to 14 days of age. UCP2 mRNA expression was not correlated with protein abundance in either breed at any age. UCP3 mRNA in AT was similar between breeds up to day 7; thereafter, expression was higher (general linear model, P<0.05) in Meishan piglets. Conversely, UCP3 mRNA expression in SM was higher in commercial piglets after day 7. Colonic temperature remained lower in Meishan than commercial piglets throughout the study; this was most obvious in the immediate post-partum period when Meishan piglets had lower (P<0.05) plasma triiodothyronine. In conclusion, we have demonstrated that porcine genotype influences the expression and abundance of UCP2 and 3, an influence which may, in part, be due to the distinctive endocrine profiles associated with each genotype. PMID- 15525581 TI - Overexpression of short heterodimer partner recovers impaired glucose-stimulated insulin secretion of pancreatic beta-cells overexpressing UCP2. AB - The short heterodimer partner (SHP) (NR0B2) is an orphan nuclear receptor whose function in pancreatic beta-cells is unclear. Mitochondrial uncoupling protein (UCP2) in beta-cells is upregulated in obesity-related diabetes, causing impaired glucose-stimulated insulin secretion (GSIS). We investigated whether SHP plays a role in UCP2-induced GSIS impairment. We overexpressed SHP in normal islet cells and in islet cells overexpressing UCP2 by an adenovirus-mediated infection technique. We found that SHP overexpression enhanced GSIS in normal islets, and restored GSIS in UCP2-overexpressing islets. SHP overexpression increased the glucose sensitivity of ATP-sensitive K+ (KATP) channels and enhanced the ATP/ADP ratio. A peroxisome proliferator-activated receptor gamma (PPARgamma) antagonist, GW9662, did not block the SHP effect on GSIS. SHP overexpression also corrected the impaired sensitivity of UCP2-overexpressing beta-cells to methylpyruvate, another energy fuel that bypasses glycolysis and directly enters the Krebs cycle. KATP channel inhibition mediated by dihydroxyacetone, which gives reducing equivalents directly to complex II of the electron transport system, was similar in Ad-Null-, Ad-UCP2- and Ad-UCP2+Ad-SHP-infected cells. The mitochondrial metabolic inhibitor sodium azide totally blocked the effect of SHP overexpression on GSIS. These results suggest that SHP positively regulates GSIS in beta-cells and restores glucose sensitivity in UCP2-overexpressing beta-cells by enhancing mitochondrial glucose metabolism, independent of PPARgamma activation. PMID- 15525582 TI - Induction of glucose transporter 1 expression through hypoxia-inducible factor 1alpha under hypoxic conditions in trophoblast-derived cells. AB - Glucose transporter 1 (GLUT1) plays an important role in the transport of glucose in the placenta. During early pregnancy, placentation occurs in a relatively hypoxic environment that is essential for appropriate embryonic development, and GLUT1 expression is enhanced in response to oxygen deficiency in the placenta. Hypoxia-inducible factor-1 (HIF-1)alpha is involved in the induction of GLUT1 expression in other cells. The present study was designed to test whether HIF 1alpha is involved in hypoxia-induced activation of GLUT1 expression using trophoblast-derived human BeWo and rat Rcho-1 cells as models. GLUT1 mRNA and protein expression were elevated under 5% O2 or in the presence of cobalt chloride, which has been shown to mimic hypoxia. Using rat GLUT1 (rGLUT1) promoter-luciferase constructs, we showed that this up-regulation was mediated at the transcriptional level. Deletion mutant analysis of the rGLUT1 promoter indicated that a 184 bp hypoxia-responsive element (HRE) of the promoter was essential to increase GLUT1 reporter gene expression in response to low-oxygen conditions. BeWo and Rcho-1 cells cultured under 5% O2 or with CoCl2 showed increased expression of HIF-1alpha protein compared with those cultured under 20% O2. To test whether this factor is directly involved in hypoxia-induced GLUT1 promoter activation, BeWo and Rcho-1 cells were transiently transfected with an HIF-1alpha expression vector. Exogeneous HIF-1alpha markedly increased the GLUT1 promoter activity from constructs containing the HRE site, while the GLUT1 promoter constructs lacking the HRE site were not activated by exogenous HIF 1alpha These data demonstrate that GLUT1 is up-regulated under 5% O2 or in the presence of CoCl2 in the placental cell lines through HIF-1alpha interaction with a consensus HRE site of the GLUT1 promoter. PMID- 15525584 TI - Hepatocyte growth factor regulates proliferation and differentiation of epithelial monolayers derived from islets of postnatal rat pancreas. AB - Hepatocyte growth factor (HGF) has been suggested to be a potent regulator of beta-cell function and proliferation. The purpose of this study was to investigate whether HGF could regulate the proliferation and differentiation of islet-derived epithelial monolayers into insulin-producing cells. We have generated islet-derived epithelial monolayers that are enriched with cells expressing c-Kit, a tyrosine kinase receptor and putative marker, from isolated postnatal rat islets. Monolayers were cultured on type I collagen gel and treated in defined differentiation medium with or without HGF (50 ng/ml) for 7 days. Subsequently, the expression of transcription factors and pancreatic endocrine cell markers as well as c-Kit expression were compared between the HGF (HGF+), no HGF treatment (HGF-) and monolayers without differentiation medium (control) groups, using immunocytochemical and RT-PCR approaches. We observed that the number of c-Kit-, glucose transport type 2 (Glut2)- and the transcription factor pancreatic duodenal homeobox-1 (PDX-1)-expressing cells were significantly increased in the HGF+ group. The expression of insulin at the mRNA and protein level was also increased in this treatment group with a 1.7-fold increase in basal insulin release and a 2.3-fold increase in insulin content in comparison with the HGF- group. A high proliferative capacity was also found in the HGF+ group. Co-localization of insulin and PDX-1 or Glut2 was revealed frequently in cells treated with HGF+ with occasional co-staining of c-Kit and insulin observed. This study showed that HGF can activate the proliferation and differentiation of islet-derived epithelial monolayer into insulin-producing cells. However, no formation of islet-like clusters was observed. Taken together, this study implies that HGF mediates differentiation of immature cell types into insulin-expressing cells; however, HGF supplementation alone is insuffcient in restoring full beta-cell function. PMID- 15525583 TI - High glucose potentiates cytokine- and streptozotocin-induced apoptosis of rat islet cells: effect on apoptosis-related genes. AB - Pancreatic beta-cell apoptosis is known to participate in the beta-cell destruction process that occurs in diabetes. A better understanding of how it takes place is essential for future development of therapeutic strategies aimed at preventing beta-cell loss and diabetes. In this study we determine the possible role that high glucose concentration might play as an enhancer of cytokine- and streptozotocin (STZ)-mediated rat islet cell apoptosis in vitro and its relationship with potential changes in the expression of pro- and anti apoptotic proteins. Rat islets treated with a cytokine combination (interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma) displayed a significant increase in islet cell apoptosis when the islets were incubated in 24.4 mM glucose compared with untreated islets at the same glucose concentration (13.07 +/- 1.78% vs 6.09 +/- 0.78%; P < 0.01) or islets incubated in 5.5 mM glucose concentration and cytokines (13.07 +/- 1.78% vs 8.04 +/- 1.56%; P < 0.05). IL-1beta alone did not induce a significant increase in the apoptotic rates in islet cells cultured at normal or high glucose concentrations. STZ significantly increased islet cell apoptosis when islets were cultured in 24.4 mM glucose concentration compared with untreated islets at the same glucose concentration (6.02 +/- 0.62% vs 4.44 +/- 0.63%; P < 0.05). High glucose induced an increase in Fas expression in the islet cells, and this increase was maintained after cytokine or STZ treatment. However, the expression of anti apoptotic mediators such as bcl-2 and bcl-xL did not show any significant change. These results suggest that cytokine- and STZ-mediated apoptotic effects on islet cells might be mediated by a glucose-induced hyperfunctional status and associated with an increase in Fas (Apo-1, CD-95) expression and no changes in the expression of the anti-apoptotic proteins bcl-xL and bcl-2. PMID- 15525585 TI - Diurnal changes in hypothalamic neuropeptide and SOCS-3 expression: effects of lactation and relationship with serum leptin and food intake. AB - Rats normally eat about 85% of their food at night. Lactation increases food intake 3- to 4-fold, but the diurnal pattern of food intake persists. The mechanisms responsible for the diurnal and lactation-induced changes in food intake are still unresolved, hence we have further investigated the possible roles of serum leptin and hypothalamic expression of neuropeptide Y (NPY), agouti related peptide (AgRP) and pro-opiomelanocortin (POMC) in rats. Suppressor of cytokine signalling-3 (SOCS-3) acts as a feedback inhibitor of leptin signalling in the hypothalamus, hence changes in expression of SOCS-3 were also investigated. Changes in expression of NPY, AgRP or POMC alone could not account for the diurnal changes in intake and their alteration by lactation. However, there were increased AgRP mRNA:POMC mRNA ratios at night and also during lactation, which were very similar to estimated changes in food intake. Such changes in expression may result in dominance of the orexigenic AgRP peptide over the appetite-suppressing POMC-derived peptides, and so could contribute to the hyperphagia in these states. Diurnal and lactation-related changes in the AgRP mRNA:POMC mRNA ratio and food intake are not due to changes in leptin alone. However, hypoleptinaemia, possibly through increased expression of NPY, may contribute to the hyperphagia of lactation. In the dark, expression of SOCS-3 was decreased in non-lactating rats; lactation decreased SOCS-3 expression in both light and dark phases. However, such changes are likely to enhance the ability of leptin-responsive neurones to transmit the leptin signal, and so are unlikely to contribute to either the nocturnal increase in appetite or the hyperphagia of lactation. PMID- 15525586 TI - Vascular endothelial growth factor is increased during early stage of diabetic nephropathy in type II diabetic rats. AB - Vascular endothelial growth factor (VEGF) has been implicated in the pathogenesis of diabetic nephropathy. We investigated serial changes of VEGF in the kidney and assessed whether glomerular and urinary VEGF levels are related to the severity of diabetic nephropathy. Furthermore, we examined the relationship between urinary VEGF levels and the urinary albumin excretion (UAE) rate in Otsuka-Long Evans-Tokushima-Fatty (OLETF) rats. Glomerular VEGF mRNA expression and protein synthesis were evaluated by the reverse transcription-polymerase chain reaction, immunohistochemical staining and in situ hybridization. Urinary levels of VEGF were determined by enzyme-linked immunosorbent assay. UAE was significantly higher in OLETF rats than in control Long-Evans-Tokushima-Fatty (LETO) rats throughout the study period. Urinary VEGF levels were significantly higher from 25 to 37 weeks, and then gradually reduced until 55 weeks, although the levels were still higher than those in control rats. Urinary VEGF levels also showed a significant positive correlation with UAE (r=0.262, P=0.045) and serum creatinine (r=0.398, P=0.044), and were found to be independently correlated with UAE by Spearman's rank correlation. By immunohistochemical staining and in situ hybridization, VEGF was mainly detected in the podocytes in the glomeruli. Interestingly, a significant increase in VEGF mRNA expression was observed in the early period of diabetic nephropathy, and this was associated with increased urinary VEGF excretion. Thus, the overproduction of VEGF in the diabetic kidney may participate in the pathogenesis of early-stage diabetic nephropathy. PMID- 15525587 TI - Growth hormone-induced blood pressure decrease is associated with increased mRNA levels of the vascular smooth muscle KATP channel. AB - Growth hormone (GH) deficiency is associated with abnormal vascular reactivity and development of atherosclerosis. GH treatment in GH deficient states restores systemic vascular resistance, arterial compliance, endothelium-dependent and endothelium-independent vasodilation, and may reverse markers of early atherosclerosis. However, very little is known about the molecular mechanisms underlying these effects. In the present study, male Sprague Dawley rats were hypophysectomized and treated for two weeks with GH (recombinant human GH, 2 mg/kg/day) or saline as s.c. injections twice daily. GH decreased aortic systolic blood pressure compared with saline-treated animals, while the diastolic blood pressure was not significantly changed. GH treatment increased cardiac output as determined by Doppler-echocardiography and the calculated systemic vascular resistance was markedly reduced. In order to identify GH-regulated genes of importance for vascular function, aortic mRNA levels were analyzed by the microarray technique and correlated to the systolic blood pressure levels. Using this approach, we identified 18 GH-regulated genes with possible impact on vascular tone and atherogenesis. In particular, mRNA levels of the inwardly rectifying potassium channel Kir6.1 and the sulfonylurea receptor 2B, which together form the vascular smooth muscle ATP-sensitive potassium channel, were both up-regulated by GH treatment and highly correlated to systolic blood pressure. Our findings establish a major role for GH in the regulation of vascular physiology and gene expression. Increased expression of the ATP sensitive potassium channel, recently shown to be crucial in the regulation of vascular tone, constitutes a possible mechanism by which GH governs vascular tone. PMID- 15525588 TI - Rosiglitazone impacts negatively on bone by promoting osteoblast/osteocyte apoptosis. AB - Thiazolidinediones (TZDs) increase peripheral tissue insulin sensitivity in patients with type 2 diabetes mellitus by activating the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma). In bone marrow stromal cell cultures and in vivo, activation of PPARgamma by high doses (20 mg/kg/day) of TZDs has been reported to alter stem cell differentiation by promoting commitment of progenitor cells to the adipocytic lineage while inhibiting osteoblastogenesis. Here, we have examined the in vivo effects of low dose rosiglitazone (3 mg/kg/day) on bone, administered to mice by gavage for 90 days. Rosiglitazone-treated mice had increased weight when compared with controls, with no significant alterations in serum levels of glucose, calcium or parathyroid hormone (PTH). Bone mineral density (BMD) at the lumbar vertebrae (L1 L4), ilium/sacrum, and total body was diminished by rosiglitazone treatment. Histologically, bone was characterized by decreased trabecular bone volume and increased marrow space with no significant change in bone marrow adipocity. Decreased osteoblast number and activity due to increased apoptotic death of osteoblasts and osteocytes was apparent while osteoclast parameters and serum levels of osteocalcin, alkaline phosphatase activity, and leptin were unaltered by rosiglitazone treatment. Therefore, the imbalance in bone remodeling that follows rosiglitazone administration arises from increased apoptotic death of osteogenic cells and diminished bone formation leading to the observed decrease in trabecular bone volume and BMD. These novel in vivo effects of TZDs on bone are of clinical relevance as patients with type 2 diabetes mellitus and other insulin resistant states treated with these agents may potentially be at increased risk of osteoporosis. PMID- 15525589 TI - Lentiviral vectors efficiently transduce human gonadotroph and somatotroph adenomas in vitro. Targeted expression of transgene by pituitary hormone promoters. AB - Despite important advances in human therapeutics, no specific treatment for both non-functioning gonadotroph and resistant somatotroph adenomas is available. Gene transfer by viral vectors can be considered as a promising way to achieve a specific and efficient treatment. Here we show the possibility of efficient gene transfer in human pituitary adenoma cells in vitro using a human immunodeficiency virus (HIV)-type 1-derived vector. Using enhanced green fluorescent protein (eGFP) gene as a marker placed under the phosphoglycerate kinase (PGK) promoter, gonadotroph and somatotroph adenomas were transduced even with moderate viral loads. The expression started at day 2, reached a peak at day 5, and it was still present at day 90. For targeting somatotroph and gonadotroph adenomas, human growth hormone (GH) promoter (GH -481, +54 bp) and two fragments of the human glycoprotein hormone alpha-subunit promoter (alpha-subunit 1 -520, +33 bp, and alpha-subunit 2 -907, +33 bp) were tested. In gonadotroph adenomas, the percentage of identified fluorescent cells and the fluorescence intensity analyzed by fluorescence-activated cell sorting indicated that the strength of the alpha-subunit 1 and alpha-subunit 2 promoters were comparable to that of the PGK promoter. Primary cultures of rat pituitary cells showed that alpha-subunit 1 is more selective to thyreotroph and gonadotroph phenotypes than alpha-subunit 2. GH promoter activity appeared weak in somatotroph adenomas. The human GH enhancer did not increase the GH promoter activity at all but the human prolactin promoter (-250 bp) allowed 4-fold more fluorescent cells to be obtained than the GH promoter. Several cell lines appeared too permissive to test cell-specificity of pituitary promoters. However, on human non-pituitary cell cultures, the tested pituitary promoters seemed clearly selective to target endocrine pituitary phenotypes. This study gives a starting point for a gene-therapy program using lentiviral vectors to transfer therapeutic genes in human pituitary adenomas. PMID- 15525590 TI - Type II iodothyronine deiodinase protein in chicken choroid plexus: additional perspectives on T3 supply in the avian brain. AB - It is widely accepted that type II iodothyronine deiodinase (D2) is mostly present in the brain, where it maintains the homeostasis of thyroid hormone (TH) levels. Although intensive studies have been performed on activity and mRNA levels of the deiodinases, very little is known about their expression at the protein level due to the lack of specific antisera. The current study reports the production of a specific D2 polyclonal antiserum and its use in the comparison of D2 protein distribution with that of type I (D1) and type III (D3) deiodinase protein in the choroid plexus at the blood-brain barrier level. Immunocytochemistry showed very high D2 protein expression in the choroid plexus, especially in the epithelial cells, whereas the D1 and D3 proteins were absent. Furthermore, dexamethasone treatment led to an up-regulation of the D2 protein in the choroid plexus. The expression of D2 protein in the choroid plexus led to a novel insight into the working mechanism of the uptake and transport of thyroid hormones along the blood-brain barrier in birds. It is hypothesized that D2 allows the prohormone thyroxine (T4) to be converted into the active 3,5,3' triiodothyronine (T3). Within the choroidal epithelial cells. T3 is subsequently bound to its carrier protein, transthyretin (TTR), to allow transport through the cerebrospinal fluid. Neurons can thus not only be provided with a sufficient T3 level via the aid of the astrocytes, as was hypothesized previously based on in situ hybridization data, but also by means of T4 deiodination by D2, directly at the blood-brain barrier level. PMID- 15525591 TI - Thyroid hormones modulate the endocrine and autocrine/paracrine actions of leptin on thyrotropin secretion. AB - We investigated the influence of hypo- and hyperthyroidism on the ability of leptin to modulate TSH secretion. Two hours after receiving leptin (8 mug leptin/100 g BW; s.c.), hyperthyroid rats (10 mug thyroxine (T4)/100 g body weight (BW) for 5 days) showed a 1.7-fold increase in serum TSH (P<0.05); in hypothyroid rats, leptin had no effect. Hemi-pituitaries of hyperthyroid rats incubated with 10(-9) and 10(-7)M leptin showed reductions in TSH release of 40 and 50% respectively (P<0.05); incubation with 1:2000 and 1:500 dilutions of antiserum against leptin resulted in 3- and 4-fold higher TSH release (P<0.05 and P<0.001 respectively). However, in hypothyroid pituitaries leptin or the antiserum had no effect. The results suggest that the in vivo and in vitro responsiveness of TSH to leptin is abolished in hypothyroidism and is preserved in short-term hyperthyroidism, in comparison to previous reports in euthyroidism. In addition, the inhibitory action of pituitary leptin is enhanced in hyperthyroid glands, which may suggest a role for locally produced leptin in the suppression of TSH release associated with hyperthyroidism. PMID- 15525592 TI - Estrogen response element and the promoter context of the human and mouse lactoferrin genes influence estrogen receptor alpha-mediated transactivation activity in mammary gland cells. AB - A critical step in estrogen action is the recognition of estrogen responsive elements (EREs) by liganded estrogen receptor. Our current studies were designed to determine whether an extended estrogen response element half-site (ERRE) contributes to the differential estrogen responses of the human and mouse lactoferrin overlapping chicken ovalbumin upstream promoter/ERE sequences (estrogen response modules, ERMs) in the context of their natural promoters. Transient transfections of MCF-7 cells show that liganded estrogen receptor alpha (ERalpha) activates transcription of the human lactoferrin ERM fourfold higher than the mouse lactoferrin ERM in the context of their natural promoters. Since the ERRE of the human lactoferrin gene naturally occurs 18 bp upstream from the ERM and is absent in the mouse lactoferrin gene promoter, we created a chimeric mouse lactoferrin CAT reporter, which now encodes the ERRE in the identical location as in the human lactoferrin gene. The addition of the ERRE in the mouse lactoferrin gene rendered this reporter extremely responsive to estrogen stimulation. Using limited protease digestions and electrophoretic mobility shift assays, we showed that the binding and protease sensitivity of ERalpha bound to the mouse ERM with or without the ERRE, differed. Importantly, occupancy of additional nuclear receptors at the ERRE may contribute to ERalpha binding and activation. Furthermore, the presence of ERRE influences the selectivity of coactivators in liganded ERalpha-mediated transcriptional activity. When the receptor is bound to human and mouse plus genes, which contain the ERRE, steroid receptor coactivator (SRC)-2 was preferred, while SRC-1 and SRC-3 coactivators selectively enhanced the mouse lactoferrin gene activity. Moreover, peroxisome proliferator activated receptor-gamma coactivator-1 (PGC-1alpha) and PGC-1 related estrogen receptor coactivator (PERC) robustly increase the transcriptional function of ERalpha in the presence of the ERRE. In conclusion, these data show that the context of the lactoferrin gene influences the ERalpha mediated transcriptional activity. PMID- 15525594 TI - Aromatase is abundantly expressed by neonatal rat penis but downregulated in adulthood. AB - Although synthesis of estrogen by male gonads has been well documented for over half a century, it is only recently that the role of estrogen in male reproductive events has gained appreciation. We recently reported abundant expression of estrogen receptor (ER)-alpha and -beta in different cell types of the rat penis, whose levels diminished with advancing age. The present study, which builds on data from the ER study, was designed to determine whether the penis is capable of generating its own local estrogen by examining evidence of the expression of aromatase, a microsomal enzymatic complex which irreversibly converts androgens to estrogens, using immunohistochemistry, Western blotting, in situ hybridization and real-time PCR analyses. Secondly, the effects of sex steroid hormones on penile aromatase were examined. Discrete aromatase immunoreactive cells were localized in primordial corpus cavernosum, corpus spongiosus and os penis, blood vessels and sensory corpuscle of glans penis. In situ hybridization signals corresponded with immunohistochemical findings. Western blot, enzyme immunoassay and real-time PCR analyses of rat penile samples revealed an age-dependent expression of aromatase and estrogen, with levels at week 1 almost resembling those of the ovary, but they decreased sharply by week 8, and decreased further by week 35. This expression pattern was strikingly similar to that of ER-alpha reported previously. Testosterone and diethylstilbesterol administered prenatally upregulate levels of aromatase mRNA and protein, and estrogen postnatally. Dihydrotestosterone upregulated aromatase mRNA and protein, but not estrogen. We conclude that estrogen acts via ER in a paracrine and/or autocrine manner to regulate penile events, particularly during development, and that estrogen synthesis is regulated by estrogen and androgens. PMID- 15525593 TI - Multiple plasma proteins control atrial natriuretic peptide (ANP) aggregation. AB - We have recently demonstrated that human alpha-atrial natriuretic peptide (alpha hANP), an amyloidogenic peptide responsible for isolated atrial amyloidosis, binds to a dimeric form of apo A-I belonging to small high-density lipoproteins (HDL). This binding phenomenon is considered a protective mechanism since it inhibits or strongly reduces the ANP aggregation process. The observation that plasma exhibits at least four times greater amyloid inhibitory activity than HDL prompted us to determine whether small HDL are the only ANP plasma-binding factors. After incubation of whole plasma with labelled ANP, the macromolecular complexes were subjected to two-dimensional gel electrophoresis followed by autoradiography. The results presented here provide novel evidence of additional binding proteins, in addition to apo A-I dimer, able to bind ANP in vitro and to prevent its aggregation. The mass spectrometry analysis of the radioactive spots identified them as albumin, alpha-1 antitrypsin, orosomucoid and apo A-IV-TTR complex. The putative impact of these findings in the amyloidogenic/antiamyloidogenic peptides network is discussed. PMID- 15525595 TI - Nuclear oxysterol receptors, LXRs, are involved in the maintenance of mouse caput epididymidis structure and functions. AB - In this study we looked at the epididymides and spermatozoa of mice knocked-out for nuclear oxysterol receptors (LXR). We have shown that LXR-deficient mice exhibited upon ageing a severe disruption of their caput epididymides associated with abnormal accumulation of neutral lipids. The epididymis defaults were correlated with sperm head fragility and infertility. In agreement with the observed caput defect in transgenic animals in which both LXRalpha and LXRbeta isoforms were disrupted, we have shown here that both receptors are expressed in caput and cauda epididymides regions. LXRbeta was predominantly expressed throughout the mouse epididymis while the expression of LXRalpha was weaker. In addition, the expression of selected genes that can be considered as markers of adult epididymis function was monitored via Northern blots in the different single and double LXR-deficient backgrounds. Altogether, the data presented here suggest that LXR receptors are important actors in epididymis function. PMID- 15525596 TI - Contributions of the N- and C-terminal domains of IGF binding protein-6 to IGF binding. AB - Insulin-like growth factors IGF-I and IGF -II are important mediators of growth. A family of six high affinity IGF binding proteins (IGFBPs) modulate IGF action. IGFBPs have three domains, of which the N- and C-domains are involved in high affinity IGF binding. IGFBP-6 is unique in its 20-100-fold IGF-II binding specificity over IGF-I. The aim of this study was to determine the contributions of the N- and C-domains of IGFBP-6 to its IGF binding properties. We confirmed that differential dissociation kinetics are responsible for the IGF-II binding preference of IGFBP-6. The N-domain has rapid association kinetics, similar to full-length IGFBP-6, but both IGF-I and -II dissociate rapidly from this domain, thereby reducing its binding affinity for IGF-II approximately 50-fold. However, the N-domain binds IGF-I and -II with similar affinities and it has a similar IGF I binding affinity to full-length IGFBP-6. This suggests that the C-domain confers the IGF-II binding preference of IGFBP-6; indeed, IGF-I bound inconsistently with very low affinity to the C-domain. Coincubation studies showed that isolated N- and C-domains of IGFBP-6 do not strongly cooperate to enhance IGF binding. The results of the binding studies are supported by the effects of the IGFBP-6 domains on IGF-induced colon cancer cell proliferation; the N-domain inhibited IGF-II induced proliferation with approximately 20-fold lower potency than IGFBP-6 and it was equipotent in inhibiting IGF-I- and IGF-II induced proliferation. Coincubation of C-domain had no additional effect on N domain-induced inhibition of proliferation. In conclusion, both the N- and C domains of IGFBP-6 are involved in IGF binding, the C-domain is responsible for the IGF-II binding preference of IGFBP-6 and intact IGFBP-6 is necessary for high affinity IGF binding. PMID- 15525597 TI - Estrogen response element-dependent regulation of transcriptional activation of estrogen receptors alpha and beta by coactivators and corepressors. AB - One mechanism by which ligand-activated estrogen receptors alpha and beta (ERalpha and ERbeta) stimulate gene transcription is through direct ER interaction with specific DNA sequences, estrogen response elements (EREs). ERE bound ER recruits coactivators that stimulate gene transcription. Binding of ER to natural and synthetic EREs with different nucleotide sequences alters ER binding affinity, conformation, and transcriptional activity, indicating that the ERE sequence is an allosteric effector of ER action. Here we tested the hypothesis that alterations in ER conformation induced by binding to different ERE sequences modulates ER interaction with coactivators and corepressors. CHO-K1 cells transfected with ERalpha or ERbeta show ERE sequence-dependent differences in the functional interaction of ERalpha and ERbeta with coactivators steroid receptor coactivator 1 (SRC-1), SRC-2 (glucocorticoid receptor interacting protein 1 (GRIP1)), SRC-3 amplified in breast cancer 1 (AIB1) and ACTR, cyclic AMP binding protein (CBP), and steroid receptor RNA activator (SRA), corepressors nuclear receptor co-repressor (NCoR) and silencing mediator for retinoid and thyroid hormone receptors (SMRT), and secondary coactivators coactivator associated arginine methyltransferase 1 (CARM1) and protein arginine methyltransferase 1 (PRMT1). We note both ligand-independent as well estradiol- and 4-hydroxytamoxifen-dependent differences in ER-coregulator activity. In vitro ER-ERE binding assays using receptor interaction domains of these coregulators failed to recapitulate the cell-based results, substantiating the importance of the full-length proteins in regulating ER activity. These data demonstrated that the ERE sequence impacts estradiol-and 4-hydroxytamoxifen-occupied ERalpha and ERbeta interaction with coregulators as measured by transcriptional activity in mammalian cells. PMID- 15525598 TI - Duplicated receptors for VIP and PACAP (VPAC1R and PAC1R) in a teleost fish, Fugu rubripes. AB - Two principal groups of receptors orthologous with human PAC1R and VPAC1R and were identified and characterised at the genomic level in the teleost fish Fugu rubripes. An additional group orthologous with VPAC2R was also identified and partially characterised. In Fugu, gene duplication of each of the PAC1Rs, VPAC1Rs and VPAC2Rs appears to have occurred. The topology of the tree surrounding the Fugu duplications and other isolated piscine sequences indicates that the duplication events for these six genes clearly preceded the speciation event leading to the Cypriniformes and Tetraodontiformes and is probably teleost specific. Overall, the combined pattern of gene expression for each pair of duplicated genes mirrored the expression in other vertebrates. However, within each pair of duplicates further specialisation had occurred, with each demonstrating differential tissue distribution profiles suggesting they that may be responsible for the divergent action of the ligands, vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP). The Fugu VPAC1R gene regions showed conserved synteny with human chromosome 3p21.3 and also C. elegans chromosome X, indicating that the putative ancestral human chromosome 3 region may be equivalent to chromosome X in Caenorhabditis elegans. PMID- 15525599 TI - Effects of dihydrotestosterone on adipose tissue measured by serial analysis of gene expression. AB - Intra-abdominal fat accumulation is related to several diseases, especially diabetes and heart disease. Molecular mechanisms associated with this independent risk factor are not well established. Through the serial analysis of gene expression (SAGE) strategy, we have studied the transcriptomic effects of castration and dihydrotestosterone (DHT) in retroperitoneal adipose tissue of C57BL6 male mice. Approximately 50,000 SAGE tags were isolated in intact and gonadectomized mice, as well as 3 and 24 h after DHT administration. Transcripts involved in energy metabolism, such as glyceraldehyde-3-phosphate dehydrogenase, malic enzyme supernatant, fatty acid synthase, lipoprotein lipase, hormone sensitive lipase and monoglyceride lipase, were upregulated by DHT. Transcripts involved in adipogenesis, and cell cycle and cell shape organization, such as DDX5, C/EBPalpha, cyclin I, procollagen types I, III, IV, V and VI, SPARC and matrix metalloproteinase 2, were upregulated by DHT. Cell defense, division and signaling, protein expression and many novel transcripts were regulated by castration and DHT. The present results provide global genomic evidence for a stimulation of glycolysis, fatty acids and triacylglycerol production, lipolysis and cell shape reorganization, as well as cell proliferation and differentiation, by DHT. The novel transcripts regulated by DHT may contribute to identify new mechanisms involved in the action of sex hormones and their potential role in obesity. PMID- 15525600 TI - Regulation of fibronectin by thyroid hormone receptors. AB - Thyroid hormones regulate growth, development, differentiation, and metabolic processes by interacting with and activating thyroid hormone receptors and associated pathways. We investigated the triiodothyronine (T3) modulation of gene expression, in human hepatocellular carcinoma cell lines, via a PCR-based cDNA subtraction method. Here we present further data on one of the T3-upregulated genes, fibronectin (FN). We demonstrate that the induction of FN protein expression by T3 in TRalpha1 and TRbeta1 over-expressing cells was time and dose dependent at the mRNA and protein levels. Blockade of protein synthesis by cycloheximide almost completely inhibited the concomitant induction of FN mRNA by T3, indicating that T3 indirectly regulates FN. Furthermore, nuclear-run on and FN promoter assay clearly can specifically increase the number of FN transcriptional demonstrated that the presence of T3 initiations. In addition, we further confirmed that the up-regulation of FN by T3 was mediated, at least in part, by transforming growth factor-beta (TGF-beta), because the induction of FN was blocked in a dose-dependent manner by the addition of TGF-beta neutralizing antibody. In an effort to elucidate the we demonstrated the involvement of the signaling pathways involved in the activation of FN by T3, mitogen activated protein kinase/c-Jun N-terminal kinase/p38 MAPK (MAPK/JNK/p38) pathway. Although T3 induces the expression of TGF-beta, neither wild-type nor dominant-negative Smad3 or Smad4 over-expression affected the activation of FN by T3. Thus, we demonstrate that T3 regulates FN gene expression indirectly at the transcriptional level, with the participation of the MAPK/JNK/p38 pathway and the TGF-beta signaling pathway but independent of Smad3/4. PMID- 15525601 TI - Localization of 17beta-hydroxysteroid dehydrogenase type 1 mRNA in mouse tissues. AB - The enzyme 17beta-hydroxysteroid dehydrogenase (17beta-HSD) type 1 catalyzes the conversion of estrone (E1) into 17beta estradiol (E2). To gain information about the cellular localization of 17beta-HSD mRNA type 1 expression, we performed in situ hybridization using a 35S-labeled cRNA probe in several tissues of adult mice of both sexes. In the ovary, high expression was found in granulosa cells of growing follicles. No specific labeling could be observed in corpora lutea or interstitial cells. In the pituitary gland of animals of both sexes, 17beta-HSD type 1 mRNA was expressed in the intermediate lobe melanotrophs while no specific signal could be detected in the anterior or posterior lobes of the pituitary. In the prostate, 17beta-HSD type 1 mRNA was exclusively found in the epithelial cells. In both male and female mouse dorsal skin, a specific hybridization signal was seen in the sebaceous glands while the epidermis, stroma, hair follicles and sweat glands were unlabeled. In the testis, a hybridization signal was detected in germ cells of the seminiferous tubules, Leydig cells being unlabeled. The present data indicate that E2 can be formed through the action of 17beta-HSD type 1 in specific cells of the gonads and peripheral tissues. In the testes and peripheral tissues, the action of E2 is probably limited to the cells involved in its formation in an intracrine fashion. PMID- 15525602 TI - Thyroid hormone signaling is highly heterogeneous during pre- and postnatal brain development. AB - We have generated transgenic reporter mice to analyze the spatio-temporal distribution of thyroid hormone signaling during mouse brain development. The reporter system, utilizing a chimeric yeast Gal4 DNA-binding domain-thyroid hormone alpha ligand-binding domain fusion protein to drive lacZ expression, revealed that thyroid hormone signaling starts in the midbrain roof several days before the onset of thyroid gland function, and that it remains highly heterogeneous in the central nervous system throughout pre- and postnatal development. We speculate that this heterogeneity might provide neural cells with positional information during development. PMID- 15525603 TI - A bioinformatics-based functional analysis shows that the specifically androgen regulated gene SARG contains an active direct repeat androgen response element in the first intron. AB - We characterized the specifically androgen-regulated gene (SARG), which is expressed in the androgen receptor (AR) and glucocorticoid receptor (GR) positive cell line lymph node carcinoma of the prostate-1F5 (LNCaP-1F5). SARG mRNA expression can be up-regulated by androgens, but not by glucocorticoids. SARG mRNA expression is high in prostate tissue. SARG is composed of four exons and spans a region of 14.5 kbp on chromosome 1q32.2. Transcripts of 5.5, 3.3 and 2.3 kb are the result of alternative polyadenylation. SARG mRNA splice variants lack exon 2 and vary in length of exon 1. The SARG protein has a length of 601 amino acids and is located in the cytoplasm. By screening the 18 kbp genomic sequence flanking the transcription start site we identified the imperfect direct repeat 5'-TGTGCTaacTGTTCT-3'in intron 1 as an active androgen response element (ARE SARG+4.6). A 569 bp genomic DNA fragment containing this element functioned as an androgen-specific enhancer in transiently transfected LNCaP-1F5 cells. ARE SARG+4.6 cooperated with flanking sequences for optimal activity. Inactivation of ARE-SARG+4.6 completely abolished the androgen response of the enhancer. Chromatin immunoprecipitation (ChIP) experiments showed chromatin structural changes of the enhancer in the presence of R1881. ARE-SARG+4.6 was able to bind to the androgen receptor, but not to the glucocorticoid receptor, correlating with its androgen-specific activity in transfections. PMID- 15525604 TI - Dimerization is required for transactivation by estrogen-receptor-related (ERR) orphan receptors: evidence from amphioxus ERR. AB - The estrogen-receptor-related (ERR) receptors are orphan members of the nuclear receptor superfamily that bind to their specific DNA target sites as homodimers. However, it has not been shown whether this mode of binding is required for the transcriptional activation they drive. We here show that heterodimerization can also occur between these receptors. Furthermore, we demonstrate that the unique amphioxus ortholog of ERR genes (AmphiERR) is expressed as two isoforms differing by an in-frame insertion. While the short isoform behaves like its mammalian counterparts, the long isoform (AmphiERR(L)) displays divergent transcriptional properties according to the target site to which it binds. Indeed, AmphiERR(L) binds as a monomer but does not activate transcription through the SF1 response element (SFRE). On the contrary, this isoform binds as a homodimer and activates transcription through the classical estrogen-response element. Our results strongly suggest that dimerization is required for transactivation exerted by the ERR receptors. PMID- 15525606 TI - Manipulating sorting signals to generate co-expression of somatostatin and eGFP in the regulated secretory pathway from a monocistronic construct. AB - Targeted overexpression of biologically active peptides represents a powerful approach to the functional dissection of neuroendocrine systems. However, the requirement to generate separate, biologically active and reporter molecules necessitates the use of internal ribosome entry site (IRES) technology, which often results in preferential translation of the second cistron. We report here a novel approach in which the proteolytic processing machinery of the regulated secretory pathway (RSP) has been exploited to generate multiple mature proteins from a monocistronic construct that encodes a single precursor. This was achieved by duplication of the pre-pro cleavage sites in pre-prosomatostatin cDNA. The duplicated site included 10 flanking amino acids on either side of the Gly-Ala cleavage position. This enabled the incorporation of a foreign protein-coding sequence (in this case, enhanced green fluorescent protein (eGFP)) between these sites. The pre-eGFP-prosomatostatin (PEPS) construct generated co-localized expression of fully processed eGFP and somatostatin to the RSP of transiently transfected AtT20 cells. This approach represents an advance upon bicistronic and other extant approaches to the targeting of multiple, biologically active proteins to neuroendocrine systems, and, importantly, permits the co-expression of fluorescent markers with biologically active neuropeptides. In this study, our demonstration of the fusion of the first 10 amino acids of the prosomatostatin sequence to the N-terminus of eGFP shows that this putative sorting sequence is sufficient to direct expression to the RSP. PMID- 15525605 TI - cAMP-dependent protein kinase activation inhibits proliferation and enhances apoptotic effect of tumor necrosis factor-alpha in NCI-H295R adrenocortical cells. AB - Adrenocorticotropin is the major regulator of adrenocortical development and function. It acts mainly through the cAMP-dependent protein kinase A (PKA) pathway. Our aim was to study the interaction of tumor necrosis factor-alpha (TNFalpha) and the PKA pathway in adrenocortical cell proliferation and apoptosis. The PKA activator Dibutyryl cAMP ((Bu)2cAMP) strongly induced differentiation and inhibited proliferation in the human adrenocortical cell line NCI-H295R (H295R). TNFalpha induced apoptosis of H295R cells. Interestingly, (Bu)2cAMP treatment clearly enhanced TNFalpha-induced apoptosis in H295R cells, but not in another human adrenocortical cell line SW-13, the mouse adrenocortical Y-1 cell line or the human HeLa cell line. This synergistic effect was not due to the (Bu)2cAMP-induced glucocorticoid secretion since dexamethasone had no significant effect on the TNFalpha-induced apoptosis. (Bu)2cAMP treatment rapidly increased the expression of the proto-oncogene c-myc in H295R cells, but not in SW-13, Y-1 or HeLa cells. In transient c-myc transfection assay, c-myc expression associated with decreased expression of the proliferation marker Ki-67 in H295R cells. In conclusion, cAMP-dependent protein kinase activation reduced proliferation and augmented TNFalpha-induced apoptosis in adrenocortical H295R cells, and these effects were associated with increased c-myc expression. PMID- 15525607 TI - Suppression of plasma free fatty acids upregulates peroxisome proliferator activated receptor (PPAR) alpha and delta and PPAR coactivator 1alpha in human skeletal muscle, but not lipid regulatory genes. AB - Fatty acids are an important ligand for peroxisome proliferator-activated receptor (PPAR) activation and transcriptional regulation of metabolic genes. To examine whether reduced plasma free fatty acid (FFA) availability affects the mRNA content of proteins involved in fuel metabolism in vivo, the skeletal muscle mRNA content of various transcription factors, transcriptional coactivators and genes encoding for lipid regulatory proteins were examined before and after 3 h of cycle exercise with (NA) and without (CON) pre-exercise ingestion of nicotinic acid (NA). NA resulted in a marked (3- to 6-fold) increase (P<0.05) in PPARalpha, PPARdelta and PPAR coactivator 1alpha (PGC1alpha) mRNA, but was without effect on nuclear respiratory factor-1 and Forkhead transcription factor, fatty acid transcolase/CD36, carnitine palmitoyl transferase 1, hormone sensitive lipase (HSL) and pyruvate dehydrogenase kinase 4. Exercise in CON was associated with increased (P<0.05) PPARalpha, PPARdelta and PGC1alpha mRNA, which was similar in magnitude to levels observed with NA at rest. Exercise was generally without effect on the mRNA content of lipid regulatory proteins in CON and did not affect the mRNA content of the measured subset of transcription factors, transcriptional co-activators and lipid regulatory proteins during NA. To determine the possible mechanisms by which NA might affect PGC1alpha expression, we measured p38 MAP kinase (MAPK) and plasma epinephrine. Phosphorylation of p38 MAPK was increased (P<0.05) by NA treatment at rest, and this correlated (r2=0.84, P<0.01) with increased PGC1alpha. Despite this close relationship, increasing p38 MAPK in human primary myotubes was without effect on PGC1alpha mRNA content. Plasma epinephrine was elevated (P<0.05) by NA at rest (CON: 0.27+/-0.06, NA: 0.72+/ 0.11 nM) and throughout exercise. Incubating human primary myotubes with epinephrine increased PGC1alpha independently of changes in p38 MAPK phosphorylation. Hence, despite the fact that NA ingestion decreased FFA availability, it promoted the induction of PPARalpha/delta and PGC1alpha gene expression to a similar degree as prolonged exercise. We suggest that the increase in PGC1alpha may be due to the elevated plasma epinephrine levels. Despite these changes in transcription factors/coactivators, the mRNA content of lipid regulatory proteins was generally unaffected by plasma FFA availability. PMID- 15525608 TI - Permission to publish case reports/case series. PMID- 15525609 TI - Causes of brain injury during orthopedic surgery. PMID- 15525610 TI - Developing yet another spinal analgesic drug? PMID- 15525611 TI - Cerebral fat embolism diagnosed by magnetic resonance imaging at one, eight, and 50 days after hip arthroplasty: a case report. AB - PURPOSE: To describe cardiovascular collapse during a cemented hip hemiarthroplasty in a patient who, despite a successful cardiopulmonary resuscitation, remained in a persistent vegetative state due to cerebral fat embolism diagnosed by magnetic resonance imaging (MRI). CLINICAL FEATURES: A 75 yr-old woman with no medical history underwent cemented hip hemiarthroplasty under spinal anesthesia for a right femoral neck fracture. Shortly after insertion of the prosthesis, a sudden oxygen desaturation, hypotension, bradycardia, and cardiac arrest occurred. The patient was successfully resuscitated, but did not regain consciousness. The patient developed high-grade fever, thrombocytopenia, anemia, and oliguria. MRI scans of the brain revealed multiple high intensity signals throughout the white matter, the basal ganglia, the cerebellum, and the brain stem. The diagnosis of fat embolism was made on the basis of clinical findings and MRI images. Although her cardiorespiratory status improved over the next week, the patient remained in a persistent vegetative state. CONCLUSION: When fat embolism is suspected, serial MRI scans of the brain should be performed to diagnose the etiology of cerebral embolism as well as to evaluate the severity of brain damage. PMID- 15525612 TI - Arousal with iv epinephrine depends on the depth of anesthesia. AB - PURPOSE: To investigate whether the depth of anesthesia affects the change in the bispectral index (BIS) caused by iv epinephrine during propofol anesthesia. METHODS: Forty women undergoing elective lower abdominal surgery received a propofol target controlled infusion (TCI) to maintain a modified Observer's Assessment of Alertness/Sedation (OAA/S) score of 2 (sedation period). Subsequently anesthesia was induced with propofol TCI 5 mug.mL(-1) and rocuronium 0.9 mg.kg(-1), and propofol continued so as to maintain general anesthesia at a BIS of 50 (general anesthesia period). Intravenous epinephrine at a dose of 10 mug.5 mL(-1) in normal saline (epinephrine group, n = 20) or normal saline 5 mL (control group, n = 20) was administered during both periods. The BIS, mean arterial pressure (MAP) and heart rate (HR) were measured immediately before, and one, two, three, four, six, eight, and ten minutes after injection. The modified OAA/S scale was evaluated during the sedation period. RESULTS: There was no significant change in the modified OAA/S scale, BIS, or hemodynamic variables compared to preinjection values during either sedation or general anesthesia in the control group. Intravenous epinephrine increased the BIS and modified OAA/S scale during sedation, but there was no increase in BIS during general anesthesia. Increases in HR and MAP were observed during both periods after iv epinephrine. CONCLUSION: Intravenous epinephrine 10 mug resulted in an arousal effect and an increase in BIS during sedation, but did not change the BIS during general anesthesia. These results suggest that the arousal effect of iv epinephrine during propofol anesthesia depends on anesthetic depth. PMID- 15525613 TI - Thirty percent of patients have moderate to severe pain 24 hr after ambulatory surgery: a survey of 5,703 patients. AB - PURPOSE: Postoperative pain is the commonest reason for delayed discharge and unanticipated hospital admission after ambulatory surgery. We investigated the severity of pain at 24 hr postoperatively and determined the most painful procedures. The need for further medical advice and clarity of postoperative analgesia instructions were also studied. METHODS: Five thousand seven hundred and three ambulatory surgical patients were telephoned 24 hr postoperatively. Patients graded their pain using the ten-point self-assessing verbal scale (0 = no pain, 10 = worst pain). Data were analyzed in two groups, those with moderate to severe pain (pain score 4-10) and those with no or mild pain (0-3). RESULTS: Thirty percent of patients (1,495/5,703) had moderate to severe pain. Microdiscectomy, laparoscopic cholecystectomy, shoulder surgery, elbow/hand surgery, ankle surgery, inguinal hernia repair, and knee surgery were identified as the procedures causing most pain at 24 hr. 13.2% of patients needed medical advice by telephone, 1.4% made an unplanned visit to a doctor while the rate of readmission to the hospital was 0.08%. Ninety-eight percent found postoperative instruction sheets and advice helpful. Eighty-eight percent of patients indicated that analgesic instructions were absolutely clear. CONCLUSION: This study has identified the more painful common ambulatory surgical procedures which will allow take home analgesia to be tailored according to individual procedures. Further improvement in analgesic instructions may help in better pain management of ambulatory surgery patients. PMID- 15525614 TI - Intrathecal propofol has analgesic effects on inflammation-induced pain in rats. AB - PURPOSE: Propofol is thought to act on gamma-aminobutyric acid receptors, which have some role in pain transmission in the spinal cord. In this study, we examined the effects of intrathecal propofol on acute thermally- or inflammation induced pain in rats. METHODS: Lumbar intrathecal catheters were implanted in Male Sprague-Dawley rats. The tail withdrawal response to thermal stimulation (tail flick test) or paw flinching and shaking response by sc formalin injection into the hind paw (formalin test) were tested. Propofol 1000, 300 or 100 microg or saline (control) was administered as 10 microL intrathecally. Motor disturbance and behavioural side effects were also monitored in the rats during the tail flick test. Eight rats were used for each dose in each test. RESULTS: No analgesic effects were observed in the tail flick test. In the formalin test, 50% of effective doses were 449 mug (95% confidence interval, 80-3180 microg) in phase 1 and 275 microg (146-519 microg) in phase 2. Motor disturbance was observed in one rat with 100 microg and agitation and allodynia were seen in one rat with 300 microg. However, both were reversible in 120 min. CONCLUSIONS: Intrathecal administration of propofol had analgesic effects on inflammation induced acute and facilitated pain but not on thermally-induced acute pain. Transient motor and sensory disturbance could not rule out the possibility of neurotoxicity. PMID- 15525615 TI - Dextromethorphan potentiates morphine antinociception at the spinal level in rats. AB - PURPOSE: Morphine is an effective analgesic, but adverse effects limit its clinical use in higher doses. The non-opioid antitussive, dextromethorphan (DM), can potentiate the analgesic effect of morphine and decrease the dose of morphine in acute postoperative pain, but the underlying mechanism remains unclear. We previously observed that DM increases the serum concentration of morphine in rats. Therefore, we investigated the effects of drugs administered at the spinal level to exclude possible pharmacokinetic interactions. As DM has widespread binding sites in the central nervous system [such as N-methyl-D-aspartate (NMDA) receptors, sigma receptors and alpha(3)ss(4) nicotinic receptors], we investigated whether the potentiation of morphine antinociception by DM at the spinal level is related to NMDA receptors. METHODS: We used MK-801 as a tool to block the NMDA channel first, and then studied the interaction between intrathecal (i.t.) morphine and DM. The tail-flick test was used to examine the antinociceptive effects of different combinations of morphine and other drugs in rats. RESULTS: DM (2-20 microg) or MK-801 (5-15 microg) showed no significant antinociceptive effect by themselves. The antinociceptive effect of morphine (0.5 microg, i.t.) was significantly enhanced by DM and reached the maximal potentiation (43.7%-50.4%) at doses of 2 to 10 microg. Pretreatment with MK-801 (5 or 10 microg, i.t.) significantly potentiated morphine antinociception by 49.9% or 38.7%, respectively. When rats were pretreated with MK-801, DM could not further enhance morphine antinociception (45.7% vs 50.5% and 43.3%). CONCLUSION: Our results suggest that spinal NMDA receptors play an important role in the effect of DM to potentiate morphine antinociception. PMID- 15525616 TI - Sufentanil modifies the antibacterial activity of bupivacaine and ropivacaine. AB - PURPOSE: The purpose of our study was to investigate the effect on the growth of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus), and Enterococcus faecalis (E. faecalis) of bupivacaine at a final concentration of 0.77 mg.mL(-1), ropivacaine at 1.2 mg.mL(-1), and sufentanil at 0.38 and 0.5 microg.mL(-1) (alone or in combination with bupivacaine and ropivacaine). METHODS: The strains were diluted to approximately 3 x 10(4) cfu.mL(-1) in Mueller-Hinton broth. The anesthetics (0.5 mL) were incubated with the bacterial suspensions (0.5 mL) for 24 hr at 37 degrees C. RESULTS: Bupivacaine inhibited the growth of E. coli (59 +/- 0.8%; P < 0.05) and S. aureus (22 +/- 3.6%; P < 0.05). Ropivacaine also inhibited the growth of E. coli (41 +/- 1.2%; P < 0.05) and S. aureus (25.5 +/- 4.1%; P < 0.05). Both anesthetics were ineffective against E. faecalis. Sufentanil only inhibited S. aureus (13.8 +/- 3.1%; P < 0.05) at a concentration of 0.5 microg.mL(-1). Sufentanil modified the antibacterial activity of bupivacaine and ropivacaine. It increased the inhibitory effect of bupivacaine on E. faecalis and S. aureus by 10 +/- 2.1% (P < 0.05) and on E. coli by 7% (P < 0.05). Sufentanil did not increase the inhibitory effect of ropivacaine on the growth of S. aureus. On the other hand, sufentanil reduced the inhibitory effect of ropivacaine on E. coli by 11% (P < 0.05). CONCLUSION: Both bupivacaine and ropivacaine alone or combined with sufentanil inhibited the growth of E. coli and S. aureus. E. faecalis was partially sensitive to a bupivacaine + sufentanil mixture. Sufentanil had a partial synergistic effect on bupivacaine and a partial antagonistic effect on ropivacaine's antibacterial activity. PMID- 15525617 TI - A modified needle-inside-needle technique for the ganglion impar block. AB - PURPOSE: The ganglion impar is the fused terminus of the paired sympathetic chain located at the level of the sacrococcygeal junction. It has been blocked using a bent and a curved spinal needle via the anococcygeal ligament. It has also been approached through the sacrococcygeal disc using a straight spinal needle. We describe a needle-inside-needle modification of the latter approach.Technical features: A 22-gauge (G), 1(1/2)-inch (38 mm) needle is introduced through the sacrococcygeal ligament under fluoroscopy via the sacrococcygeal disc. A 25-G, 2 inch (50 mm) needle is introduced through the 22-G needle. Placement is confirmed with injection of iopamidol 300, 0.2 mL in the retroperitoneal space with the comma sign. CONCLUSIONS: The bent and curved needle techniques are associated with significant discomfort, tissue trauma and risk of rectal perforation due to difficulty in obtaining a midline needle tip position. The straight spinal needle approach minimizes these problems, however there is increased risk of discitis and a longer spinal needle may help also raise incidence of needle breakage. The needle-inside-needle technique may reduce these risks. PMID- 15525619 TI - Anesthetic management of Cesarean delivery in a patient with hypoplastic anemia and severe pre-eclampsia. AB - PURPOSE: To describe the anesthetic management of Cesarean delivery in a patient with hypoplastic anemia and severe pre-eclampsia. CLINICAL FEATURES: A 28-yr-old parturient with a history of thrombocytopenia was admitted with signs of pre eclampsia (blood pressure of 140/90 mmHg, heavy proteinuria and moderate bilateral ankle edema) at 25 weeks of gestation. Laboratory studies revealed pancy-topenia (hemoglobin 6.4 g.dL(-1), white cell count 3.43 x 10(9).L(-1), platelet count 20 x 10(9).L(-1)) and bone marrow biopsy showed hypoplastic anemia. As pre-eclampsia worsened, a Cesarean delivery was performed at 27 weeks with prophylactic platelet transfusion and meticulous blood pressure control. The procedure was uneventful, conducted under general anesthesia with an estimated blood loss of around 600 mL and a live female baby was delivered. Postoperatively her blood pressure and neurological symptoms improved but thrombocytopenia remained at discharge. CONCLUSIONS: Hypoplastic anemia is rare in pregnancy but it poses an increased risk for both mother and fetus. The mother is at risk of life-threatening episodes of bleeding and infection and a multidisciplinary team approach (obstetrician, anesthesiologist, hematologist and pediatrician) is essential. An accurate assessment of the hematological condition should be made and abnormalities corrected before surgery. Regional anesthesia may not be possible in this circumstance. PMID- 15525618 TI - 0.2% Ropivacaine and levobupivacaine provide equally effective epidural labour analgesia. AB - PURPOSE: To compare the duration of epidural analgesia induced by levobupivacaine and ropivacaine at clinically relevant doses. METHODS: Forty healthy nulliparous parturients with cervical dilatation of 3 to 5 cm and at least one contraction every two or three minutes were recruited. Patients were randomly assigned to receive either 10 mL of 0.2% ropivacaine or 10 mL of 0.2% levobupivacaine. Preblock visual analogue scale (VAS) score (0-100) and VAS score after five, ten, 15, 20, 25 and 30 min from time(0) and VAS at time of request for additional analgesia (time(end)) were recorded. During the first 30 min after the completion of epidural injection, the systolic blood pressure, highest sensory level to cold and the maximum degree of motor block based on a 0 to 3 modified Bromage scale were collected every five minutes. The duration of analgesia was defined as the time from time(0) to time(end). RESULTS: There was no difference in the duration of analgesia between the two groups; similarly, there was no significant difference found in the area under the curve (AUC) time(15)-time(0) and AUC time(30)-time(0) for VAS. The highest sensory block to cold and the degree of motor block were also indistinguishable between the two groups. No difference in the serial systolic blood pressures was found. CONCLUSION: Ten millilitres of either 0.2% ropivacaine or levobupivacaine can be used to induce epidural labour analgesia effectively without a difference in the duration of pain relief. PMID- 15525620 TI - Patient-controlled regional analgesia is effective in children: a preliminary report. AB - PURPOSE: To report a preliminary analysis of prospectively recorded data in 27 children in whom patient-controlled regional analgesia (PCRA) was used for postoperative pain control following lower limb surgery. METHODS: Under general anesthesia, perineural catheters (popliteal and fascia iliaca compartment block) were inserted and infused with ropivacaine 0.2% (0.02 mL.kg(-1).hr(-1)). Additional demand doses were left to the child's discretion (0.1 mL.kg(-1)and a 30-min lockout interval). RESULTS: The average total dose of ropivacaine administered was 4.9 +/- 2 mg.kg(-1)over 48 hr. Visual analogue scale and Children's Hospital of Eastern Ontario Pain Scale scores were always inferior to 5/10 and 6/13, respectively. Motor block was observed in two children and two children needed rescue analgesia. CONCLUSIONS: Our preliminary observations indicate that PCRA in children provides satisfactory postoperative pain relief following lower limb surgery. PMID- 15525621 TI - Images in Anesthesia: transesophageal echocardiography enhances endovascular stent placement in traumatic trans-section of the thoracic aorta. PMID- 15525622 TI - Gargling with povidone-iodine reduces the transport of bacteria during oral intubation. AB - PURPOSE: Nosocomial pneumonia remains a common complication in patients undergoing endotracheal intubation. This study examined the transport of bacteria into the trachea during endotracheal intubation, and evaluated the effects of gargling with povidone-iodine on bacterial contamination of the tip of the intubation tube. METHODS: In the gargling group, patients gargled with 25 mL of povidone-iodine (2.5 mg.mL(-1)). In the control group, patients gargled with 25 mL of tap water. Before tracheal intubation, microorganisms were obtained from the posterior wall of the patient's pharynx using sterile cotton swabs. After anesthesia, all patients were extubated and bacteria contaminating the tip of the tracheal tube were sampled and cultured. RESULTS: Before orotracheal intubation, all 19 patients who gargled with tap water (control group) had bacterial colonization on the posterior walls of the pharynx. This group included five patients who had methicillin-resistant staphylococcus aureus (MRSA) in their nasal cavity preoperatively and MRSA was also detected in the pharynx of four patients. Bacterial colonization was observed in all 19 patients who gargled with povidone-iodine (gargling group) and four patients carried MRSA in their nasal cavity, although no MRSA was detected in the pharynx. In the control group, all the patients had bacterial colonization at the tip of the tube after extubation. Additionally, MRSA was detected in two of the four patients. In the gargling group, povidone-iodine eradicated general bacteria and MRSA colonies in the pharynx before intubation and at the tip of the tube after extubation. CONCLUSION: Gargling with povidone-iodine before oral intubation reduces the transport of bacteria into the trachea. PMID- 15525623 TI - Posterior tibial nerve and median nerve somatosensory evoked potential monitoring during carotid endarterectomy. AB - PURPOSE: Somatosensory evoked potential (SSEP) monitoring using the median nerve (MN) modality during carotid endarterectomy is well established. This study assessed the usefulness of monitoring the posterior tibial nerve (PTN) SSEP as an adjunct to MNSSEP for detection of cerebral ischemia and as an indicator for the insertion of a shunt in patients undergoing a carotid endarterectomy. METHODS: All patients undergoing carotid endarterectomy during three years who had routine bilateral MNSSEP were also monitored with bilateral PTNSSEP. Patients received a shunt if there was a significant change (> 50% decrease in amplitude of cortical peak (N20) in the MNSSEP after cross clamping. The incidence, timing, and duration of all PTNSSEP changes were compared to MNSSEP changes. RESULTS: One hundred fifty-three patients were studied. Significant changes in MNSSEP after cross clamp lead to insertion of a shunt in six patients. Changes in PTNSSEP occurred at almost the same time in three patients, four minutes before MNSSEP in one, three minutes later in one and no change in one patient. Good quality baseline tracings were obtained in 99% MNSSEP as compared to 88% PTNSSEP (P < 0.05). New postoperative neurological deficits occurred in four patients (2.6%), only one had significant evoked potential changes. CONCLUSION: Monitoring of PTNSSEP is feasible and may be considered for an adjunct to MNSSEP or as an alternative modality if there are difficulties with MNSSEP. However, there may be a greater incidence of poor quality baseline tracings for PTNSSEP. PMID- 15525624 TI - More on lumbo-sacral spine surgery and bradycardia. PMID- 15525625 TI - Difficulties in anesthetizing a 106-yr-old patient for total hip replacement. PMID- 15525627 TI - Early blood-patch for spontaneous intracranial hypotension. PMID- 15525626 TI - Neuroendocrine stress response in laparoscopic surgery for benign ovarian cyst. PMID- 15525628 TI - Anesthetic management of laparoscopic surgery for twin to twin transfusion syndrome. PMID- 15525629 TI - Anesthetic management of a patient with Cantrell's pentalogy diagnosed prenatally. PMID- 15525630 TI - Pediatric class zero airway. PMID- 15525631 TI - An ounce of prevention worth a pound of cure. PMID- 15525632 TI - Use of the LMA in an unusual "impossible to ventilate" situation. PMID- 15525633 TI - Continuous low-dose diclofenac infusion for fever control in patients with acute neurological lesions. PMID- 15525635 TI - Zinc transporters, ZnT5 and ZnT7, are required for the activation of alkaline phosphatases, zinc-requiring enzymes that are glycosylphosphatidylinositol anchored to the cytoplasmic membrane. AB - Numerous proteins are properly folded by binding with zinc during their itinerary in the biosynthetic-secretory pathway. Several transporters have been implicated in the zinc entry into secretory compartments from cytosol, but their precise roles are poorly understood. We report here that two zinc transporters (ZnT5 and ZnT7) localized in the secretory apparatus are responsible for loading zinc to alkaline phosphatases (ALPs) that are glycosylphosphatidylinositol-anchored membrane proteins exposed to the extracellular site. Disruption of the ZnT5 gene in DT40 cells decreased the ALP activity to 45% of that in the wild-type cells. Disruption of the ZnT7 gene lowered the ALP activity only by 20%. Disruption of both genes markedly decreased the ALP activity to <5%. Overexpression of human ZnT5 or ZnT7 in DT40 cells deficient in both ZnT5 and ZnT7 genes recovered the ALP activity to the level comparable to that in the wild-type cells. The inactive ALP protein in DT40 cells deficient in both ZnT5 and ZnT7 genes was transported to cytoplasmic membrane like the active ALP protein in the wild-type cells. Thus both ZnT5 and ZnT7 contribute to the conversion of apo-ALP to holo-ALP. PMID- 15525634 TI - TCF-4 mediates cell type-specific regulation of proglucagon gene expression by beta-catenin and glycogen synthase kinase-3beta. AB - The proglucagon gene (glu) encodes glucagon, expressed in pancreatic islets, and the insulinotropic hormone GLP-1, expressed in the intestines. These two hormones exert critical and opposite effects on blood glucose homeostasis. An intriguing question that remains to be answered is whether and how glu gene expression is regulated in a cell type-specific manner. We reported previously that the glu gene promoter in gut endocrine cell lines was stimulated by beta-catenin, the major effector of the Wnt signaling pathway, whereas glu mRNA expression and GLP 1 synthesis were activated via inhibition of glycogen synthase kinase-3beta, the major negative modulator of the Wnt pathway (Ni, Z., Anini, Y., Fang, X., Mills, G. B., Brubaker, P. L., & Jin, T. (2003) J. Biol. Chem. 278, 1380-1387). We now show that beta-catenin and the glycogen synthase kinase-3beta inhibitor lithium do not activate glu mRNA or glu promoter expression in pancreatic cell lines. In the intestinal GLUTag cell line, but not in the pancreatic InR1-G9 cell line, the glu promoter G2 enhancer-element was activated by lithium treatment via a TCF binding motif. TCF-4 is abundantly expressed in the gut but not in pancreatic islets. Furthermore, both TCF-4 and beta-catenin bind to the glu gene promoter, as detected by chromatin immunoprecipitation. Finally, stable introduction of dominant-negative TCF-4 into the GLUTag cell line repressed basal glu mRNA expression and abolished the effect of lithium on glu mRNA expression and GLP-1 synthesis. We have therefore identified a unique mechanism that regulates glu expression in gut endocrine cells only. Tissue-specific expression of TCF factors thus may play a role in the diversity of the Wnt pathway. PMID- 15525636 TI - The Neisseria meningitidis outer membrane lipoprotein FrpD binds the RTX protein FrpC. AB - At conditions of low iron availability, Neisseria meningitidis produces a family of FrpC-like, type I-secreted RTX proteins of unknown role in meningococcal lifestyle. It is shown here that iron starvation also induces production of FrpD, the other protein expressed from a gene located immediately upstream of the frpC gene in a predicted iron-regulated frpDC operon. We found that FrpD is highly conserved in a set of meningococcal strains representative of all serogroups and does not exhibit any similarity to known sequences of other organisms. Subcellular localization and [3H]palmitic acid labeling in Escherichia coli revealed that FrpD is synthesized with a type II signal peptide for export across the cytoplasmic membrane and is, upon processing to a lipoprotein, sorted to the outer bacterial membrane. Furthermore, the biological function of FrpD appears to be linked to that of the RTX protein FrpC, because FrpD was found to bind the amino-proximal portion of FrpC (first 300 residues) with very high affinity (apparent Kd approximately 0.2 nM). These results suggest that FrpD represents an rtx loci-encoded accessory lipoprotein that could be involved in anchoring of the secreted RTX protein to the outer bacterial membrane. PMID- 15525637 TI - Anthraquinones inhibit tau aggregation and dissolve Alzheimer's paired helical filaments in vitro and in cells. AB - The abnormal aggregation of tau protein into paired helical filaments (PHFs) is one of the hallmarks of Alzheimer's disease. Aggregation takes place in the cytoplasm and could therefore be cytotoxic for neurons. To find inhibitors of PHF aggregation we screened a library of 200,000 compounds. The hits found in the PHF inhibition assay were also tested for their ability to dissolve preformed PHFs. The results were obtained using a thioflavin S fluorescence assay for the detection and quantification of tau aggregation in solution, a tryptophan fluorescence assay using tryptophan-containing mutants of tau, and confirmed by a pelleting assay and electron microscopy of the products. Here we demonstrate the feasibility of the approach with several compounds from the family of anthraquinones, including emodin, daunorubicin, adriamycin, and others. They were able to inhibit PHF formation with IC50 values of 1-5 microm and to disassemble preformed PHFs at DC50 values of 2-4 microm. The compounds had a similar activity for PHFs made from different tau isoforms and constructs. The compounds did not interfere with the stabilization of microtubules by tau. Tau-inducible neuroblastoma cells showed the formation of tau aggregates and concomitant cytotoxicity, which could be prevented by inhibitors. Thus, small molecule inhibitors could provide a basis for the development of tools for the treatment of tau pathology in AD and other tauopathies. PMID- 15525638 TI - Calpain and other cytosolic proteases can contribute to the degradation of retro translocated prion protein in the cytosol. AB - PrP, a cell surface-localized N-linked glycoprotein, is required for the pathogenesis of prion diseases. Recent studies have revealed that prion protein (PrP) becomes neurotoxic and prone to aggregation when it is in the cytosol, suggesting that cytosolic PrP may play a role in the pathogenesis of prion disease. Retro-translocation of PrP from the endoplasmic reticulum to the cytosol for proteasome degradation offers a natural route for PrP to enter the cytosol, but whether PrP is subject to retrotranslocation is controversial. In this study, we investigated the metabolism of endogenous wild-type PrP in several cell lines and in primary mouse cortical neurons. Our results suggest that a portion of the endogenous wild-type PrP is retro-translocated to the cytosol and degraded by the proteasome. Moreover, we also found that calpain and other cytosolic proteases could degrade PrP in the cytosol when the proteasome activity is compromised. These results provide the foundation for the hypothesis that cytosolic PrP may be involved in the pathogenesis of prion disease. PMID- 15525639 TI - INSL5 is a high affinity specific agonist for GPCR142 (GPR100). AB - Insulin-like peptide 5 (INSL5) is a peptide that belongs to the relaxin/insulin family, and its receptor has not been identified. In this report, we demonstrate that INSL5 is a specific agonist for GPCR142. Human INSL5 displaces the binding of (125)I-relaxin-3 to GPCR142 with a high affinity (K(i) = 1.5 nM). In a saturation binding assay, (125)I-INSL5 binds GPCR142 with a K(d) value of 2.5 nM. In functional guanosine (gamma-thio)-triphosphate binding and cAMP accumulation assays, INSL5 potently activates GPCR142 with EC(50) values of 1.3 and 1.2 nM, respectively. In addition, INSL5 stimulates Ca(2+) mobilization in HEK293 cells expressing GPCR142 and G alpha(16). Overall, INSL5 behaves as an agonist for GPCR142 similar to relaxin-3. However, unlike relaxin-3, which is also a potent agonist for GPCR135 and LGR7, INSL5 does not activate either GPCR135 or LGR7. INSL5 inhibits (125)I-relaxin-3 binding to GPCR135 with a low potency (K(i) = 500 nM). A functional assay shows that INSL5 (1 microm) is a weak antagonist for GPCR135. In addition, INSL5 (up to 1 microm) shows no affinity or activity at LGR7 or LGR8 either in a binding assay or a bio-functional assay. Previously, we have demonstrated that GPCR142 mRNA is expressed in peripheral tissues, particularly in the colon. Here we show that INSL5 mRNA is expressed in many peripheral tissues, similar to GPCR142. The high affinity interaction between INSL5 and GPCR142 coupled with their co-evolution and partially overlapping tissue expression patterns strongly suggest that INSL5 is an endogenous ligand for GPCR142. PMID- 15525640 TI - Identification and characterization of a novel vitamin B12 (cobalamin) biosynthetic enzyme (CobZ) from Rhodobacter capsulatus, containing flavin, heme, and Fe-S cofactors. AB - One of the most intriguing steps during cobalamin (vitamin B12) biosynthesis is the ring contraction process that leads to the extrusion of one of the integral macrocyclic carbon atoms from the tetrapyrrole-derived framework. The aerobic cobalamin pathway requires the action of a monooxygenase called CobG (precorrin 3B synthase), which generates a hydroxylactone intermediate that is subsequently ring-contracted by CobJ. However, in the photosynthetic bacterium Rhodobacter capsulatus, which harbors an aerobic-like pathway, there is no cobG in the main cobalamin biosynthetic operon although it does contain an additional uncharacterized gene called orf663. To demonstrate the involvement of Orf663 in cobalamin synthesis, the first dedicated 10 genes of the B12 pathway (including orf663), encoding enzymes for the transformation of uroporphyrinogen III into hydrogenobyrinic acid (HBA), were sequentially cloned into a plasmid to generate an artificial operon, which, when transformed into Escherichia coli, endowed the host with the ability to make HBA. Deletion of orf663 from this operon prevented HBA synthesis, demonstrating that it was essential for corrin construction. HBA synthesis was restored to this recombinant strain either by returning orf663 or by substituting it with cobG. Recombinant overproduction of Orf663, now renamed CobZ, allowed the characterization of a novel cofactor-rich protein, housing two Fe-S centers, a flavin, and a heme group, which like B12 itself is a modified tetrapyrrole. A mechanism for Orf663 (CobZ) in cobalamin biosynthesis is proposed. PMID- 15525641 TI - Heterogeneous nuclear ribonucleoprotein A1 is a novel internal ribosome entry site trans-acting factor that modulates alternative initiation of translation of the fibroblast growth factor 2 mRNA. AB - Alternative initiation of translation of the human fibroblast growth factor 2 (FGF-2) mRNA at five in-frame CUG or AUG translation initiation codons requires various RNA cis-acting elements, including an internal ribosome entry site (IRES). Here we describe the purification of a trans-acting factor controlling FGF-2 mRNA translation achieved by several biochemical purification approaches. We have identified the heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) as a factor that binds to the FGF-2 5'-leader RNA and that also complements defective FGF-2 translation in vitro in rabbit reticulocyte lysate. Recombinant hnRNP A1 stimulates in vitro translation at the four IRES-dependent initiation codons but has no effect on the cap-dependent initiation codon. Consistent with a role of hnRNP A1 in the control of alternative initiation of translation, short interfering RNA-mediated knock down of hnRNP A1 specifically inhibits translation at the four IRES-dependent initiation codons. Furthermore, hnRNP A1 binds to the FGF-2 IRES, implicating this interaction in the control of alternative initiation of translation. PMID- 15525642 TI - The latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus modulates cellular gene expression and protects lymphoid cells from p16 INK4A induced cell cycle arrest. AB - Latently infected Kaposi's sarcoma-associated herpes-virus (KSHV)-associated tumor cells have both endothelial and lymphoid origins and express a limited set of latent viral genes. One such gene, ORF73, encodes the latency-associated nuclear antigen (LANA), a multifunctional protein that plays roles in viral DNA replication, episome maintenance, and transcriptional regulation. LANA interacts with cellular proteins involved in transcriptional regulation such as the tumor suppressors, retinoblastoma (Rb) and p53, and RING3 family members. Although several reports about specific LANA-regulated promoters exist, only limited data are available that address how LANA expression in KSHV-infected cells globally affects cellular gene expression, thereby potentially contributing to KSHV pathogenicity. To investigate this question, we generated an Epstein-Barr virus negative Burkitts lymphoma line that expresses LANA from a tetracycline-inducible promoter (BJAB/Tet-On/LANA), and we performed microarray-based gene expression profiling. Expression profiling at different time points post-induction revealed that 186 genes were activated or repressed over 2-fold in the presence of LANA. Of these genes, 41 are regulated in the Rb/E2F pathway, whereas 7 are related to p53 signaling. To determine whether these gene expression changes translate into LANA-dependent changes in cell cycle regulation, we overexpressed p16 INK4a, a CDK4/6 inhibitor that efficiently induces cell cycle arrest in Rb-positive cells. Under these conditions, LANA expression protects lymphoid cells from p16 INK4a induced cell cycle arrest and induces S-phase entry. PMID- 15525643 TI - Regiospecificity determinants of human heme oxygenase: differential NADPH- and ascorbate-dependent heme cleavage by the R183E mutant. AB - The ability of the human heme oxygenase-1 (hHO-1) R183E mutant to oxidize heme in reactions supported by either NADPH-cytochrome P450 reductase or ascorbic acid has been compared. The NADPH-dependent reaction, like that of wild-type hHO-1, yields exclusively biliverdin IXalpha. In contrast, the R183E mutant with ascorbic acid as the reductant produces biliverdin IXalpha (79 +/- 4%), IXdelta (19 +/- 3%), and a trace of IXbeta. In the presence of superoxide dismutase and catalase, the yield of biliverdin IXdelta is decreased to 8 +/- 1% with a corresponding increase in biliverdin IXalpha. Spectroscopic analysis of the NADPH dependent reaction shows that the R183E ferric biliverdin complex accumulates, because reduction of the iron, which is required for sequential iron and biliverdin release, is impaired. Reversal of the charge at position 183 makes reduction of the iron more difficult. The crystal structure of the R183E mutant, determined in the ferric and ferrous-NO bound forms, shows that the heme primarily adopts the same orientation as in wild-type hHO-1. The structure of the Fe(II).NO complex suggests that an altered active site hydrogen bonding network supports catalysis in the R183E mutant. Furthermore, Arg-183 contributes to the regiospecificity of the wild-type enzyme, but its contribution is not critical. The results indicate that the ascorbate-dependent reaction is subject to a lower degree of regiochemical control than the NADPH-dependent reaction. Ascorbate may be able to reduce the R183E ferric and ferrous dioxygen complexes in active site conformations that cannot be reduced by NADPH-cytochrome P450 reductase. PMID- 15525644 TI - Analysis of transport activity of Arabidopsis sugar alcohol permease homolog AtPLT5. AB - The transporters responsible for sugar uptake into non-photosynthetic sink tissues in plants, such as roots and flowers, have not been fully identified and analyzed. Plants encode around 100 putative sugar transporters within the major facilitator superfamily, yet only a few have been studied. Here we report the analysis of a sugar alcohol permease homolog (AtPLT5, At3g18830) from Arabidopsis. A wide range of sugars including hexoses, pentoses, tetroses, a sugar acid, and sugar alcohols but not disaccharides induced inward currents in oocytes expressing AtPLT5. AtPLT5 expression also resulted in 14C-labeled substrate uptake in oocytes, indicating that AtPLT5 encodes an ion-coupled uptake transporter. K(0.5) values for glucose and sorbitol were highly dependent on external pH. Expression of AtPLT5 was found primarily in sink tissues: in the elongation zone of roots, in the inflorescence stem, and several floral structures, especially in the floral abscission zone. Expression was induced by mechanical wounding and insect feeding. Analysis of transport properties and expression in Arabidopsis indicate that AtPLT5 functions to transport a wide range of sugars into specific sink tissues in the plant. PMID- 15525645 TI - Protein stoichiometry of a multiprotein complex, the human spliceosomal U1 small nuclear ribonucleoprotein: absolute quantification using isotope-coded tags and mass spectrometry. AB - The human U1 snRNP (small nuclear ribonucleoprotein), which is a part of the spliceosome, consists of U1 snRNA and ten different proteins: seven Sm proteins B/B', D1, D2, D3, E, F, and G and the three U1-specific proteins U1-70 K, U1-A, U1-C. To determine the stoichiometry of all ten proteins, the complex was denatured, digested completely with an endoproteinase and labeled with an amine specific tag. Corresponding peptides were synthesized and labeled with the same tag containing heavier isotopes. The digest was then spiked with defined amounts of the synthetic peptides, and the resulting isotopic peptide pairs were analyzed quantitatively by mass spectrometry. The mass spectra provided information about the absolute amount of each component in the starting protein mixture. The use of the isotope-coded, amine-specific reagents propionyl-N-oxysuccinimide and nicotinoyl-N-oxysuccinimide was evaluated for stoichiometry determination; the nicotinoyl reagent was found to be advantageous because of its greater mass spectrometric sensitivity. Absolute quantities of all ten proteins were measured, showing equal numbers of all ten proteins in the U1 spliceosomal snRNP. These data demonstrate that quantitative mass spectrometry has great potential for the determination of the stoichiometry of multiprotein complexes. PMID- 15525646 TI - Structural basis of constitutive activity and a unique nucleotide binding mode of human Pim-1 kinase. AB - Pim-1 kinase is a member of a distinct class of serine/threonine kinases consisting of Pim-1, Pim-2, and Pim-3. Pim kinases are highly homologous to one another and share a unique consensus hinge region sequence, ER-PXPX, with its two proline residues separated by a non-conserved residue, but they (Pim kinases) have <30% sequence identity with other kinases. Pim-1 has been implicated in both cytokine-induced signal transduction and the development of lymphoid malignancies. We have determined the crystal structures of apo Pim-1 kinase and its AMP-PNP (5'-adenylyl-beta,gamma-imidodiphosphate) complex to 2.1-angstroms resolutions. The structures reveal the following. 1) The kinase adopts a constitutively active conformation, and extensive hydrophobic and hydrogen bond interactions between the activation loop and the catalytic loop might be the structural basis for maintaining such a conformation. 2) The hinge region has a novel architecture and hydrogen-bonding pattern, which not only expand the ATP pocket but also serve to establish unambiguously the alignment of the Pim-1 hinge region with that of other kinases. 3) The binding mode of AMP-PNP to Pim-1 kinase is unique and does not involve a critical hinge region hydrogen bond interaction. Analysis of the reported Pim-1 kinase-domain structures leads to a hypothesis as to how Pim kinase activity might be regulated in vivo. PMID- 15525648 TI - Human immunodeficiency virus protease inhibitors accumulate into cultured human adipocytes and alter expression of adipocytokines. AB - Lipodystrophic syndrome is a major side effect of highly active antiviral therapy. Fat tissue redistribution is associated with changes in adipocyte gene expression and in circulating levels of adipocytokines involved in the development of insulin resistance. However, the evidence that HIV drugs accumulate into human adipocytes and have a direct effect on the expression of adipocyte-specific genes is still lacking. To address these questions, we used adipocytes derived from adult stem (hMADS) cells isolated from human adipose tissue. We showed by ELISA that two inhibitors of the HIV protease, lopinavir and ritonavir, accumulated at similar levels during the development of hMADS cells in adipocytes, whereas a non-nucleoside reverse transcriptase inhibitor, the nevirapine, accumulated at lower levels. Two fluorescent protease inhibitors then have been generated to investigate their subcellular localization. The data showed that HIV drugs accumulated into adipocytes and displayed various effects on hMADS cell-derived adipocytes. Indinavir, amprenavir, and nevirapine did not alter differentiation of precursor cells. In contrast, lopinavir, saquinavir, and ritonavir inhibited the development of preadipocytes into adipocytes. In adipocytes, amprenavir increased leptin expression and ritonavir was able to up regulate tumor necrosis factor-alpha, interleukin 6, and leptin expression and down-regulate the expression of peroxisome proliferator-activated receptor gamma and adiponectin. Intracellular accumulation and localization of HIV drugs into human adipocytes strongly suggest that adipose tissues store these drugs. Because ritonavir can alter the expression of insulin resistance-related cytokines in human adipocytes in a way parallel to the situation observed in vivo upon treatment of HIV-infected patients, we propose that protease inhibitors participate in insulin resistance through a direct effect on adipocytes. PMID- 15525647 TI - Arabidopsis HAF2 gene encoding TATA-binding protein (TBP)-associated factor TAF1, is required to integrate light signals to regulate gene expression and growth. AB - Plant growth and development are sensitive to light. Light-responsive DNA-binding transcription factors have been functionally identified. However, how transcription initiation complex integrates light signals from enhancer-bound transcription factors remains unknown. In this work, we characterized mutations within the Arabidopsis HAF2 gene encoding TATA-binding protein-associated factor TAF1 (or TAF(II)250). The mutation of HAF2 induced decreases on chlorophyll accumulation, light-induced mRNA levels, and promoter activity. Genetic analysis indicated that HAF2 is involved in the pathways of both red/far-red and blue light signals. Double mutants between haf2-1 and hy5-1, a mutation of a light signaling positive DNA-binding transcription factor gene, had a synergistic effect on photomorphogenic traits and light-activated gene expression under different light wavelengths, suggesting that HAF2 is required for interaction with additional light-responsive DNA-binding transcription factors to fully respond to light induction. Chromatin immunoprecipitation assays showed that the mutation of HAF2 reduced acetylation of histone H3 in light-responsive promoters. In addition, transcriptome analysis showed that the mutation altered the expression of about 9% of genes in young leaves. These data indicate that TAF1 encoded by the Arabidopsis HAF2 gene functions as a coactivator capable of integrating light signals and acetylating histones to activate light-induced gene transcription. PMID- 15525649 TI - Angiotensin II and epidermal growth factor induce cyclooxygenase-2 expression in intestinal epithelial cells through small GTPases using distinct signaling pathways. AB - Colorectal carcinogenesis is a multistep process involving genetic mutations and alterations in rigorously controlled signaling pathways and gene expression that control intestinal epithelial cell proliferation, differentiation, and apoptosis. Cyclooxygenase-2 (COX-2) is aberrantly expressed in premalignant adenomatous polyps and colorectal carcinomas and is associated with increased epithelial cell proliferation, decreased apoptosis, and increased cell invasiveness. Currently, knowledge of the regulation of expression of COX-2 by endogenous cell-surface receptors is inadequate. Recently, in a non-transformed rat intestinal epithelial cell line (IEC-18), we showed induction of cell proliferation and DNA synthesis by angiotensin II (Ang II) via the endogenous Ang II type 1 receptor (Chiu, T., Santiskulvong, C., and Rozengurt, E. (2003) Am. J. Physiol. 285, G1-G11). We report that Ang II potently stimulated expression of COX-2 mRNA and protein as an immediate-early gene response through the Ang II type 1 receptor, correlating with an increase in prostaglandin I2 production. Ang II induced Cdc42 activation and filopodial formation. COX-2 expression was induced by epidermal growth factor (EGF), which activated Rac with lamellipodial formation. Inhibition of small GTPases by Clostridium difficile toxin B blocked COX-2 expression by Ang II and EGF. Inhibition of ERK activation by U0126 or PD98059 significantly decreased EGF dependent COX-2 expression, but did not affect Ang II-dependent COX-2 expression. Conversely, inhibition of p38MAPK by SB202190 or PD169316 inhibited COX-2 expression by Ang II, but did not block COX-2 induction by EGF. Ang II caused Ca2+ mobilization. Inhibition of Ca2+ signaling by 2-aminobiphenyl borate blocked Ang II-dependent COX-2 expression. EGF did not induce Ca2+ mobilization, and 2 aminobiphenyl borate did not inhibit EGF-dependent COX-2 expression. Inhibition of COX-2 expression correlated with inhibition of prostaglandin I2 production. Luciferase promoter assays showed that Ang II-dependent transcriptional activation of the COX-2 promoter was dependent on activation of small GTPases and p38(MAPK) and on Ca2+ signaling via the cAMP-responsive element/activating transcription factor cis-acting element. PMID- 15525650 TI - The V-ATPase subunit C binds to polymeric F-actin as well as to monomeric G-actin and induces cross-linking of actin filaments. AB - Previously, we have shown that the V-ATPase holoenzyme as well as the V1 complex isolated from the midgut of the tobacco hornworm (Manduca sexta) exhibits the ability of binding to actin filaments via the V1 subunits B and C (Vitavska, O., Wieczorek, H., and Merzendorfer,H. (2003) J. Biol. Chem. 278, 18499-18505). Since the recombinant subunit C not only enhances actin binding of the V1 complex but also can bind separately to F-actin, we analyzed the interaction of recombinant subunit C with actin. We demonstrate that it binds not only to F-actin but also to monomeric G-actin. With dissociation constants of approximately 50 nm, the interaction exhibits a high affinity, and no difference could be observed between binding to ATP-G-actin or ADP-G-actin, respectively. Unlike other proteins such as members of the ADF/cofilin family, which also bind to G- as well as to F actin, subunit C does not destabilize actin filaments. On the contrary, under conditions where the disassembly of F-actin into G-actin usually occurred, subunit C stabilized F-actin. In addition, it increased the initial rate of actin polymerization in a concentration-dependent manner and was shown to cross-link actin filaments to bundles of varying thickness. Apparently bundling is enabled by the existence of at least two actin-binding sites present in the N- and in the C-terminal halves of subunits C, respectively. Since subunit C has the possibility to dimerize or even to oligomerize, spacing between actin filaments could be variable in size. PMID- 15525651 TI - Galpha12 directly interacts with PP2A: evidence FOR Galpha12-stimulated PP2A phosphatase activity and dephosphorylation of microtubule-associated protein, tau. AB - The Galpha(12/13) family of heterotrimeric G proteins modulate multiple cellular processes including regulation of the actin cytoskeleton. Galpha(12/13) interact with several cytoskeletal/scaffolding proteins, and in a yeast two-hybrid screen with Galpha(12), we detected an interaction with the scaffolding subunit (Aalpha) of the Ser/Thr phosphatase, protein phosphatase 2A (PP2A). PP2A dephosphorylates multiple substrates including tau, a microtubule-associated protein that is hyperphosphorylated in neurofibrillary tangles. The interaction of Aalpha and Galpha(12) was confirmed by coimmunoprecipitation studies in transfected COS cells and by glutathione S-transferase (GST)-Galpha(12) pull-downs from cell lysates of primary neurons. The interaction was specific for Aalpha and Galpha(12) and was independent of Galpha(12) conformation. Endogenous Aalpha and Galpha(12) colocalized by immunofluorescent microscopy in Caco-2 cells and in neurons. In vitro reconstitution of GST-Galpha(12) or recombinant Galpha(12) with PP2A core enzyme resulted in approximately 300% stimulation of PP2A activity that was not detected with other Galpha subunits and was similar with GTPgammaS- and GDP-liganded Galpha(12). When tau and active kinase (Cdk5 and p25) were cotransfected in to COS cells, there was robust tau phosphorylation. Co expression of wild type or QLalpha(12) with tau and the active kinase resulted in 60 +/- 15% reductions in tau phosphorylation. In primary cortical neurons stimulated with lysophosphatitic acid, a 50% decrease in tau phosphorylation was observed. The Galpha(12) effect on tau phosphorylation was inhibited by the PP2A inhibitor, okadaic acid (50 nm), in COS cells and neurons. Taken together, these findings reveal novel, direct regulation of PP2A activity by Galpha(12) and potential in vivo modulation of PP2A target proteins including tau. PMID- 15525652 TI - Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain. AB - Processing of the laminin-5 (Ln-5) gamma 2 chain by membrane-type-1 matrix metalloproteinases (MT1-MMP) promotes migration and invasion of epithelial and tumor cells. We previously demonstrated that MT1-MMP cleaves the rat gamma 2 chain at two sites, producing two major C-terminal fragments of 100 (gamma 2') and 80 (gamma 2 x) kDa and releasing a 30-kDa fragment containing epidermal growth factor (EGF)-like motifs (domain III (DIII) fragment). The DIII fragment bound the EGF receptor (EGF-R) and stimulated cell scattering and migration. However, it is not yet clear whether human Ln-5 is processed in a similar fashion to rat Ln-5 because one of the two MT1-MMP cleavage sites present in rat gamma 2 is not found in human gamma 2. To identify the exact cleavage site for MT1-MMP in human Ln-5, we purified both the whole molecule as well as a monomeric form of human gamma 2 that is frequently expressed by malignant tumor cells. Like rat Ln 5, both the monomer of gamma 2, as well as the gamma 2 derived from intact Ln-5, were cleaved by MT1-MMP in vitro, generating C-terminal gamma 2' (100 kDa) and gamma 2 x (85 kDa) fragments and releasing DIII fragments (25 and 27k Da). In addition to the conserved first cleavage site used to generate gamma 2', two adjacent cleavage sites (Gly(559)-Asp(560) and Gly(579)-Ser(580)) were found that could generate the gamma 2 x and DIII fragments. Two of the three EGF-like motifs present in the rat DIII fragment are present in the 27-kDa human fragment, and like the rat DIII, this fragment can promote breast carcinoma cell migration by engaging the EGF-R. These results suggest that MT1-MMP processing of Ln-5 in human tumors may stimulate the EGF-R, resulting in increased tumor cell scattering and migration that could possibly increase their metastatic potential. PMID- 15525653 TI - Integrin-mediated adhesion and soluble ligand binding stabilize COX-2 protein levels in endothelial cells by inducing expression and preventing degradation. AB - Cyclooxygenase-2 (COX-2), a key enzyme in prostaglandin synthesis, is highly expressed during inflammation and cellular transformation and promotes tumor progression and angiogenesis. We have previously demonstrated that endothelial cell COX-2 is required for integrin alphaVbeta3-dependent activation of Rac-1 and Cdc-42 and for endothelial cell spreading, migration, and angiogenesis (Dormond, O., Foletti, A., Paroz, C., and Ruegg, C. (2001) Nat. Med. 7, 1041-1047; Dormond, O., Bezzi, M., Mariotti, A., and Ruegg, C. (2002) J. Biol. Chem. 277, 45838 45846). In this study, we addressed the question of whether integrin-mediated cell adhesion may regulate COX-2 expression in endothelial cells. We report that cell detachment from the substrate caused rapid degradation of COX-2 protein in human umbilical vein endothelial cells (HUVEC) independent of serum stimulation. This effect was prevented by broad inhibition of cellular proteinases and by neutralizing lysosomal activity but not by inhibiting the proteasome. HUVEC adhesion to laminin, collagen I, fibronectin, or vitronectin induced rapid COX-2 protein expression with peak levels reached within 2 h and increased COX-2 dependent prostaglandin E2 production. In contrast, nonspecific adhesion to poly L-lysine was ineffective in inducing COX-2 expression. Furthermore, the addition of matrix proteins in solution promoted COX-2 protein expression in suspended or poly-L-lysine-attached HUVEC. Adhesion-induced COX-2 expression was strongly suppressed by pharmacological inhibition of c-Src, phosphatidylinositol 3-kinase, p38, extracellular-regulated kinase 1/2, and, to a lesser extent, protein kinase C and by the inhibition of mRNA or protein synthesis. In conclusion, this work demonstrates that integrin-mediated cell adhesion and soluble integrin ligands contribute to maintaining COX-2 steady-state levels in endothelial cells by the combined prevention of lysosomal-dependent degradation and the stimulation of mRNA synthesis involving multiple signaling pathways. PMID- 15525654 TI - Are occupational therapists more effective than social workers when assessing frail older people? Results of CAMELOT, a randomised controlled trial. AB - OBJECTIVES: To compare the effectiveness of occupational therapist-led assessments of older people on dependency and service costs with that of social worker-led assessments. DESIGN: Pragmatic community-based randomised controlled trial over 2 years 4 months. SETTING: Cambridgeshire, UK. PARTICIPANTS: 321 older people aged 65 and over living in their own homes and 113 carers. INTERVENTION: participants were randomised to two groups, to receive either occupational therapist-led or social worker-led assessment. OUTCOME MEASURES: Primary outcome was dependency (Community Dependency Index). Secondary outcomes included quality of life scores (EQ-5D) and psychological outlook (Perceived Stress Scale (PSS)). Outcome measures for carers included Carer Assessment of Difficulty Index (CADI), PSS and EQ-5D, collected at baseline, 4 and 8 months. Resource use data were collected from professional practice records, participants and carers at final follow-up. RESULTS: 264 (82%) of the randomised participants completed the study. No between-group statistically significant differences were found, except that carers in the occupational therapist arm had significantly better EQ-5D scores at the 8 month follow-up (thermometer P = 0.03) and in the social worker arm better CADI scores on stress (P = 0.047) and amount of caring (P = 0.049). CONCLUSIONS: There was no clear difference in patient-centred effectiveness measures between occupational therapists and social workers in assessing frail older people and their carers in the community. More extensive use of primary care health services by occupational therapists may have contributed to the differences in EQ-5D scores for carers. Delays in making occupational therapy assessments and in completing recommended housing adaptations may have contributed to these negative findings. PMID- 15525655 TI - The effectiveness of a modified version of the Wilkinson questionnaire in screening for TIA and minor stroke in the United Kingdom. AB - BACKGROUND: Transient ischaemic attacks (TIAs) and minor strokes are important to diagnose as there are effective secondary preventive interventions. Significant under-reporting by patients occurs, but general practitioners tend to over diagnose the condition, contributing in part to long clinic waiting lists. An accurate screening test could address both these problems. METHODS: A modified version of a questionnaire designed to detect TIA was tested against the gold standard of specialist diagnosis in two vascular outpatient clinics in Leicester, UK. RESULTS: The questionnaire was sent by post with the clinic appointment and completed by 136 participants. In 99 cases the same questionnaire was administered by a clinic nurse. Overall levels of agreement (kappa, 95% CI) with specialist diagnosis of TIA, stroke or neither were 0.32 (0.15, 0.48) and 0.31 (0.12, 0.50) for postal and administered questionnaires, respectively. When the diagnoses of TIA and stroke were combined, agreement rose to 0.38 (0.23, 0.53) for postal and 0.38 (0.20, 0.57) for administered versions. For this outcome, the postal version had a sensitivity of 0.56 (0.43, 0.68) and specificity of 0.81 (0.71, 0.90). Equivalent figures for administered questionnaires were 0.61 (0.46, 0.76) and 0.76 (0.63, 0.87). CONCLUSION: The questionnaire has potential in prioritising outpatient referrals but is not sufficiently specific to be used for research or population screening. PMID- 15525656 TI - Comprehensive identification and characterization of diallelic insertion-deletion polymorphisms in 330 human candidate genes. AB - Despite being the second most frequent type of polymorphism in the genome, diallelic insertion-deletion polymorphisms (indels) have received far less attention in the study of sequence variation. In this report, we describe an approach that can detect indels in the heterozygous state and can comprehensively identify indels in the target sequence. Using this approach, we identified 2393 indels in a set of 330 candidate genes, i.e. an average of seven indels per gene with about two indels per gene being common (minor allele frequency >or=0.1). We compared the population genetic characteristics of indels with substitutions in this data. Our data supported the findings that deletions occur more frequently in the human genome. 5'-UTR and coding regions of the genes showed a significantly lower diversity for indels compared with other regions, suggesting differences in effects of selection on indels and substitutions. Sequence diversity and pairwise linkage disequilibrium (LD) findings of the different populations were similar to earlier results and included a greater skew towards low-frequency variants and a faster rate of LD decay in the African-descent population compared with the non-African populations. Within populations, the allele frequency spectra and LD-decay profiles for indels were similar to substitutions. Overall, the findings suggest that, although the mechanisms giving rise to indels may be different from those causing substitutions, the evolutionary histories of indels and substitutions are similar, and that indels can play a valuable role in association studies and marker selection strategies. PMID- 15525657 TI - Deficiency of pantothenate kinase 2 (Pank2) in mice leads to retinal degeneration and azoospermia. AB - Pantothenate kinase-associated neurodegeneration (PKAN, formerly known as Hallervorden-Spatz syndrome) is a rare but devastating neurodegenerative disorder, resulting from an inherited defect in coenzyme A biosynthesis. As pathology in the human condition is limited to the central nervous system, specifically the retina and globus pallidus, we have generated a mouse knock-out of the orthologous murine gene (Pank2) to enhance our understanding of the mechanisms of disease and to serve as a testing ground for therapies. Over time, the homozygous null mice manifest retinal degeneration, as evidenced by electroretinography, light microscopy and pupillometry response. Specifically, Pank2 mice show progressive photoreceptor decline, with significantly lower scotopic a- and b-wave amplitudes, decreased cell number and disruption of the outer segment and reduced pupillary constriction response when compared with those of wild-type littermates. Additionally, the homozygous male mutants are infertile due to azoospermia, a condition that was not appreciated in the human. Arrest occurs in spermiogenesis, with complete absence of elongated and mature spermatids. In contrast to the human, however, no changes were observed in the basal ganglia by MRI or by histological exam, nor were there signs of dystonia, even after following the mice for one year. Pank2 mice are 20% decreased in weight when compared with their wild-type littermates; however, dysphagia was not apparent. Immunohistochemistry shows staining consistent with localization of Pank2 to the mitochondria in both the retina and the spermatozoa. PMID- 15525659 TI - Expression of cardiac myosin-binding protein-C (cMyBP-C) in Drosophila as a model for the study of human cardiomyopathies. AB - Mutations in the MYBPC3 gene encoding human cardiac myosin-binding protein-C (cMyBP-C) are associated with familial hypertrophic cardiomyopathy (FHC), but the molecular mechanisms involved are not fully understood. In addition, development of FHC is sensitive to genetic background, and the search for candidate modifier genes is crucial with a view to proposing diagnosis and exploring new therapies. We used Drosophila as the model to investigate the in vivo consequences of human cMyBP-C mutations. We first produced transgenic flies that specifically express human wild-type or two C-terminal truncated cMyBP-Cs in indirect flight muscles (IFM), a tissue particularly amenable to genetic and molecular analyses. First, incorporation of human cMyBP-C into the IFM led to sarcomeric structural abnormalities and to a flightless phenotype aggravated by age and human gene dosage. Second, transcriptome analysis of transgenic IFM using nylon microarrays showed the remodelling of a transcriptional program involving 97 out of 3570 Drosophila genes. Among them, the Calmodulin gene encoding a key component of muscle contraction, found up-regulated in transgenic IFM, was evaluated as a potential modifier gene. Calmodulin mutant alleles rescued the flightless phenotype, and therefore behave as dominant suppressors of the flightless phenotype suggesting that Calmodulin might be a modifier gene in the context of human FHC. In conclusion, we suggest that the combination of heterologous transgenesis and transcriptome analysis in Drosophila could be of great value as a way to glean insights into the molecular mechanisms underlying FHC and to propose potential candidate modifier genes. PMID- 15525658 TI - Orexin loss in Huntington's disease. AB - Huntington's disease (HD) is a devastating neurodegenerative disorder caused by an expanded CAG repeat in the gene encoding huntingtin, a protein of unknown function. Mutant huntingtin forms intracellular aggregates and is associated with neuronal death in select brain regions. The most studied mouse model (R6/2) of HD replicates many features of the disease, but has been reported to exhibit only very little neuronal death. We describe for the first time a dramatic atrophy and loss of orexin neurons in the lateral hypothalamus of R6/2 mice. Importantly, we also found a significant atrophy and loss of orexin neurons in Huntington patients. Like animal models and patients with impaired orexin function, the R6/2 mice were narcoleptic. Both the number of orexin neurons in the lateral hypothalamus and the levels of orexin in the cerebrospinal fluid were reduced by 72% in end-stage R6/2 mice compared with wild-type littermates, suggesting that orexin could be used as a biomarker reflecting neurodegeneration. Our results show that the loss of orexin is a novel and potentially very important pathology in HD. PMID- 15525660 TI - Recessive mutations in PTHR1 cause contrasting skeletal dysplasias in Eiken and Blomstrand syndromes. AB - Eiken syndrome is a rare autosomal recessive skeletal dysplasia. We identified a truncation mutation in the C-terminal cytoplasmic tail of the parathyroid hormone (PTH)/PTH-related peptide (PTHrP) type 1 receptor (PTHR1) gene as the cause of this syndrome. Eiken syndrome differs from Jansen and Blomstrand chondrodysplasia and from enchondromatosis, which are all syndromes caused by PTHR1 mutations. Notably, the skeletal features are opposite to those in Blomstrand chondrodysplasia, which is caused by inactivating recessive mutations in PTHR1. To our knowledge, this is the first description of opposite manifestations resulting from distinct recessive mutations in the same gene. PMID- 15525661 TI - Association of DJ-1 and parkin mediated by pathogenic DJ-1 mutations and oxidative stress. AB - The identification of rare monogenic forms of Parkinson's disease (PD) has provided tremendous insight into the molecular pathogenesis of this disorder. Heritable mutations in alpha-synuclein, parkin, DJ-1 and PINK1 cause familial forms of PD. In the more common sporadic form of PD, oxidative stress and derangements in mitochondrial complex-I function are considered to play a prominent role in disease pathogenesis. However, the relationship of DJ-1 with other PD-linked genes and oxidative stress has not been explored. Here, we show that pathogenic mutant forms of DJ-1 specifically but differentially associate with parkin, an E3 ubiquitin ligase. Chemical cross-linking shows that pathogenic DJ-1 mutants exhibit impairments in homo-dimer formation, suggesting that parkin may bind to monomeric DJ-1. Parkin fails to specifically ubiquitinate and enhance the degradation of L166P and M26I mutant DJ-1, but instead promotes their stability in cultured cells. The interaction of parkin with L166P DJ-1 may involve a larger protein complex that contains CHIP and Hsp70, perhaps accounting for the lack of parkin-mediated ubiquitination. Oxidative stress also promotes an interaction between DJ-1 and parkin, but this does not result in the ubiquitination or degradation of DJ-1. Parkin-mediated alterations in DJ-1 protein stability may be pathogenically relevant as DJ-1 levels are dramatically increased in the detergent-insoluble fraction from sporadic PD/DLB brains, but are reduced in the insoluble fraction from parkin-linked autosomal recessive juvenile-onset PD brains. These data potentially link DJ-1 and parkin in a common molecular pathway at multiple levels that may have important implications for understanding the pathogenesis of inherited and sporadic PD. PMID- 15525662 TI - Six1 promotes a placodal fate within the lateral neurogenic ectoderm by functioning as both a transcriptional activator and repressor. AB - Cranial placodes, which give rise to sensory organs in the vertebrate head, are important embryonic structures whose development has not been well studied because of their transient nature and paucity of molecular markers. We have used markers of pre-placodal ectoderm (PPE) (six1, eya1) to determine that gradients of both neural inducers and anteroposterior signals are necessary to induce and appropriately position the PPE. Overexpression of six1 expands the PPE at the expense of neural crest and epidermis, whereas knock-down of Six1 results in reduction of the PPE domain and expansion of the neural plate, neural crest and epidermis. Using expression of activator and repressor constructs of six1 or co expression of wild-type six1 with activating or repressing co-factors (eya1 and groucho, respectively), we demonstrate that Six1 inhibits neural crest and epidermal genes via transcriptional repression and enhances PPE genes via transcriptional activation. Ectopic expression of neural plate, neural crest and epidermal genes in the PPE demonstrates that these factors mutually influence each other to establish the appropriate boundaries between these ectodermal domains. PMID- 15525663 TI - Compartmentalization of visual centers in the Drosophila brain requires Slit and Robo proteins. AB - Brain morphogenesis depends on the maintenance of boundaries between populations of non-intermingling cells. We used molecular markers to characterize a boundary within the optic lobe of the Drosophila brain and found that Slit and the Robo family of receptors, well-known regulators of axon guidance and neuronal migration, inhibit the mixing of adjacent cell populations in the developing optic lobe. Our data suggest that Slit is needed in the lamina to prevent inappropriate invasion of Robo-expressing neurons from the lobula cortex. We show that Slit protein surrounds lamina glia, while the distal cell neurons in the lobula cortex express all three Drosophila Robos. We examine the function of these proteins in the visual system by isolating a novel allele of slit that preferentially disrupts visual system expression of Slit and by creating transgenic RNA interference flies to inhibit the function of each Drosophila Robo in a tissue-specific fashion. We find that loss of Slit or simultaneous knockdown of Robo, Robo2 and Robo3 causes distal cell neurons to invade the lamina, resulting in cell mixing across the lamina/lobula cortex boundary. This boundary disruption appears to lead to alterations in patterns of axon navigation in the visual system. We propose that Slit and Robo-family proteins act to maintain the distinct cellular composition of the lamina and the lobula cortex. PMID- 15525664 TI - Twisted gastrulation promotes BMP signaling in zebrafish dorsal-ventral axial patterning. AB - In vertebrates and invertebrates, the bone morphogenetic protein (BMP) signaling pathway patterns cell fates along the dorsoventral (DV) axis. In vertebrates, BMP signaling specifies ventral cell fates, whereas restriction of BMP signaling by extracellular antagonists allows specification of dorsal fates. In misexpression assays, the conserved extracellular factor Twisted gastrulation (Tsg) is reported to both promote and antagonize BMP signaling in DV patterning. To investigate the role of endogenous Tsg in early DV patterning, we performed morpholino (MO)-based knockdown studies of Tsg1 in zebrafish. We found that loss of tsg1 results in a moderately strong dorsalization of the embryonic axis, suggesting that Tsg1 promotes ventral fates. Knockdown of tsg1 combined with loss of function of the BMP agonist tolloid (mini fin) or heterozygosity for the ligand bmp2b (swirl) enhanced dorsalization, supporting a role for Tsg1 in specifying ventral cell fates as a BMP signaling agonist. Moreover, loss of tsg1 partially suppressed the ventralized phenotypes of mutants of the BMP antagonists Chordin or Sizzled (Ogon). Our results support a model in which zebrafish Tsg1 promotes BMP signaling, and thus ventral cell fates, during DV axial patterning. PMID- 15525665 TI - Focal adhesion kinase is not required for integrin function or viability in Drosophila. AB - The mammalian focal adhesion kinase (FAK) family of non-receptor protein-tyrosine kinases has been implicated in controlling a multitude of cellular responses to the engagement of cell-surface integrins and G-protein-coupled receptors. The high level of sequence conservation between the mammalian proteins and the Drosophila homologue of FAK, Fak56, suggested that it would have similar functions. However, we show here that Drosophila Fak56 is not essential for integrin functions in adhesion, migration or signaling in vivo. Furthermore, animals lacking Fak56 are viable and fertile, demonstrating that Fak56 is not essential for other developmental or physiological functions. Despite this, overexpressed Fak56 is a potent inhibitor of integrins binding to the extracellular matrix, suggesting that Fak56 may play a subtle role in the negative regulation of integrin adhesion. PMID- 15525666 TI - Temporal complexity within a translational control element in the nanos mRNA. AB - Translational control of gene expression plays a fundamental role in the early development of many organisms. In Drosophila, selective translation of nanos mRNA localized to the germ plasm at the posterior of the embryo, together with translational repression of nanos in the bulk cytoplasm, is essential for development of the anteroposterior body pattern. We show that both components to spatial control of nanos translation initiate during oogenesis and that translational repression is initially independent of Smaug, an embryonic repressor of nanos. Repression during oogenesis and embryogenesis are mediated by distinct stem loops within the nanos 3' untranslated region; the Smaug-binding stem-loop acts strictly in the embryo, whereas a second stem-loop functions in the oocyte. Thus, independent regulatory modules with temporally distinct activities contribute to spatial regulation of nanos translation. We propose that nanos evolved to exploit two different stage-specific translational regulatory mechanisms. PMID- 15525668 TI - Evolutionary diversification of specification mechanisms within the O/P equivalence group of the leech genus Helobdella. AB - Developmental fates and cell lineage patterns are highly conserved in the teloblast lineages that give rise to the segmental ectoderm of clitellate annelids. But previous studies have shown that the pathways involved in specification of the ventrolateral O lineage and the dorsolateral P lineage differ to some degree in distantly related clitellate species such as the leeches Helobdella and Theromyzon, and the sludgeworm Tubifex. To examine this developmental variation at a lower taxonomic level, we have explored the specification pathways of the O and P lineages in the leech genus Helobdella. In leech, the O and P lineages arise from a developmental equivalence group of O/P teloblasts. In this study, we demonstrate that the cell-cell interactions involved in cell fate specification of the O/P equivalence group differ among three laboratory colonies of closely related species. In two populations, the Q lineage is necessary to specify the P fate in the dorsalmost O/P lineage, but in the third population the P fate can be specified by a redundant pathway involving the M lineage. We also observe interspecific variation in the role played by cell interactions within the O/P equivalence group, and in the apparent significance of extrinsic signals from the micromere cell lineages. Our data suggest that cell fate specification in the O/P equivalence group is a complex process that involves multiple cell-cell interactions, and that the developmental architecture of the O/P equivalence group has undergone evolutionary diversification in closely related species, despite maintaining a conserved morphology. PMID- 15525667 TI - Stabilization of beta-catenin in the mouse zygote leads to premature epithelial mesenchymal transition in the epiblast. AB - Many components of the Wnt/beta-catenin signaling pathway are expressed during mouse pre-implantation embryo development, suggesting that this pathway may control cell proliferation and differentiation at this time. We find no evidence for a functional activity of this pathway in cleavage-stage embryos using the Wnt reporter line, BAT-gal. To further probe the activity of this pathway, we activated beta-catenin signaling by mating a zona pellucida3-cre (Zp3-cre) transgenic mouse line with a mouse line containing an exon3-floxed beta-catenin allele. The result is expression of a stabilized form of beta-catenin, resistant to degradation by the GSK3beta-mediated proteasome pathway, expressed in the developing oocyte and in each cell of the resulting embryos. Nuclear localization and signaling function of beta-catenin were not observed in cleavage-stage embryos derived from these oocytes. These results indicate that in pre implantation embryos, molecular mechanisms independent of the GSK3beta-mediated ubiquitination and proteasome degradation pathway inhibit the nuclear function of beta-catenin. Although the mutant blastocysts initially developed normally, they then exhibited a specific phenotype in the embryonic ectoderm layer of early post implantation embryos. We show a nuclear function of beta-catenin in the mutant epiblast that leads to activation of Wnt/beta-catenin target genes. As a consequence, cells of the embryonic ectoderm change their fate, resulting in a premature epithelial-mesenchymal transition. PMID- 15525669 TI - Independent roles of the dachshund and eyes absent genes in BMP signaling, axon pathfinding and neuronal specification. AB - In the Drosophila nerve cord, a subset of neurons expresses the neuropeptide FMRFamide related (Fmrf). Fmrf expression is controlled by a combinatorial code of intrinsic factors and an extrinsic BMP signal. However, this previously identified code does not fully explain the regulation of Fmrf. We have found that the Dachshund (Dac) and Eyes Absent (Eya) transcription co-factors participate in this combinatorial code. Previous studies have revealed an intimate link between Dac and Eya during eye development. Here, by analyzing their function in neurons with multiple phenotypic markers, we demonstrate that they play independent roles in neuronal specification, even within single cells. dac is required for high level Fmrf expression, and acts potently together with apterous and BMP signaling to trigger Fmrf expression ectopically, even in motoneurons. By contrast, eya regulates Fmrf expression by controlling both axon pathfinding and BMP signaling, but cannot trigger Fmrf ectopically. Thus, we show that dac and eya perform entirely different functions in a single cell type to ultimately regulate a single phenotypic outcome. PMID- 15525670 TI - Formation of membrane-bound ring complexes by prohibitins in mitochondria. AB - Prohibitins comprise a remarkably conserved protein family in eukaryotic cells with proposed functions in cell cycle progression, senescence, apoptosis, and the regulation of mitochondrial activities. Two prohibitin homologues, Phb1 and Phb2, assemble into a high molecular weight complex of approximately 1.2 MDa in the mitochondrial inner membrane, but a nuclear localization of Phb1 and Phb2 also has been reported. Here, we have analyzed the biogenesis and structure of the prohibitin complex in Saccharomyces cerevisiae. Both Phb1 and Phb2 subunits are targeted to mitochondria by unconventional noncleavable targeting sequences at their amino terminal end. Membrane insertion involves binding of newly imported Phb1 to Tim8/13 complexes in the intermembrane space and is mediated by the TIM23 translocase. Assembly occurs via intermediate-sized complexes of approximately 120 kDa containing both Phb1 and Phb2. Conserved carboxy-terminal coiled-coil regions in both subunits mediate the formation of large assemblies in the inner membrane. Single particle electron microscopy of purified prohibitin complexes identifies diverse ring-shaped structures with outer dimensions of approximately 270 x 200 angstroms. Implications of these findings for proposed cellular activities of prohibitins are discussed. PMID- 15525671 TI - Actin and septin ultrastructures at the budding yeast cell cortex. AB - Budding yeast has been a powerful model organism for studies of the roles of actin in endocytosis and septins in cell division and in signaling. However, the depth of mechanistic understanding that can be obtained from such studies has been severely hindered by a lack of ultrastructural information about how actin and septins are organized at the cell cortex. To address this problem, we developed rapid-freeze and deep-etch techniques to image the yeast cell cortex in spheroplasted cells at high resolution. The cortical actin cytoskeleton assembles into conical or mound-like structures composed of short, cross-linked filaments. The Arp2/3 complex localizes near the apex of these structures, suggesting that actin patch assembly may be initiated from the apex. Mutants in cortical actin patch components with defined defects in endocytosis disrupted different stages of cortical actin patch assembly. Based on these results, we propose a model for actin function during endocytosis. In addition to actin structures, we found that septin-containing filaments assemble into two kinds of higher order structures at the cell cortex: rings and ordered gauzes. These images provide the first high resolution views of septin organization in cells. PMID- 15525672 TI - Analysis of a spindle pole body mutant reveals a defect in biorientation and illuminates spindle forces. AB - The spindle pole body (SPB) is the microtubule organizing center in Saccharomyces cerevisiae. An essential task of the SPB is to ensure assembly of the bipolar spindle, which requires a proper balancing of forces on the microtubules and chromosomes. The SPB component Spc110p connects the ends of the spindle microtubules to the core of the SPB. We previously reported the isolation of a mutant allele spc110-226 that causes broken spindles and SPB disintegration 30 min after spindle formation. By live cell imaging of mutant cells with green fluorescent protein (GFP)-Tub1p or Spc97p-GFP, we show that spc110-226 mutant cells have early defects in spindle assembly. Short spindles form but do not advance to the 1.5-microm stage and frequently collapse. Kinetochores are not arranged properly in the mutant cells. In 70% of the cells, no stable biorientation occurs and all kinetochores are associated with only one SPB. Examination of the SPB remnants by electron microscopy tomography and fluorescence microscopy revealed that the Spc110-226p/calmodulin complex is stripped off of the central plaque of the SPB and coalesces to from a nucleating structure in the nucleoplasm. The central plaque components Spc42p and Spc29p remain behind in the nuclear envelope. The delamination is likely due to a perturbed interaction between Spc42p and Spc110-226p as detected by fluorescence resonance energy transfer analysis. We suggest that the force exerted on the SPB by biorientation of the chromosomes pulls the Spc110-226p out of the SPB; removal of force exerted by coherence of the sister chromatids reduced fragmentation fourfold. Removal of the forces exerted by the cytoplasmic microtubules had no effect on fragmentation. Our results provide insights into the relative contributions of the kinetochore and cytoplasmic microtubules to the forces involved in formation of a bipolar spindle. PMID- 15525673 TI - The A78V mutation in the Mad3-like domain of Schizosaccharomyces pombe Bub1p perturbs nuclear accumulation and kinetochore targeting of Bub1p, Bub3p, and Mad3p and spindle assembly checkpoint function. AB - During mitosis, the spindle assembly checkpoint (SAC) responds to faulty attachments between kinetochores and the mitotic spindle by imposing a metaphase arrest until the defect is corrected, thereby preventing chromosome missegregation. A genetic screen to isolate SAC mutants in fission yeast yielded point mutations in three fission yeast SAC genes: mad1, bub3, and bub1. The bub1 A78V mutant is of particular interest because it produces a wild-type amount of protein that is mutated in the conserved but uncharacterized Mad3-like region of Bub1p. Characterization of mutant cells demonstrates that the alanine at position 78 in the Mad3-like domain of Bub1p is required for: 1) cell cycle arrest induced by SAC activation; 2) kinetochore accumulation of Bub1p in checkpoint-activated cells; 3) recruitment of Bub3p and Mad3p, but not Mad1p, to kinetochores in checkpoint-activated cells; and 4) nuclear accumulation of Bub1p, Bub3p, and Mad3p, but not Mad1p, in cycling cells. Increased targeting of Bub1p-A78V to the nucleus by an exogenous nuclear localization signal does not significantly increase kinetochore localization or SAC function, but GFP fused to the isolated Bub1p Mad 3-like accumulates in the nucleus. These data indicate that Bub1p-A78V is defective in both nuclear accumulation and kinetochore targeting and that a threshold level of nuclear Bub1p is necessary for the nuclear accumulation of Bub3p and Mad3p. PMID- 15525674 TI - Staufen recruitment into stress granules does not affect early mRNA transport in oligodendrocytes. AB - Staufen is a conserved double-stranded RNA-binding protein required for mRNA localization in Drosophila oocytes and embryos. The mammalian homologues Staufen 1 and Staufen 2 have been implicated in dendritic RNA targeting in neurons. Here we show that in rodent oligodendrocytes, these two proteins are present in two independent sets of RNA granules located at the distal myelinating processes. A third kind of RNA granules lacks Staufen and contains major myelin mRNAs. Myelin Staufen granules associate with microfilaments and microtubules, and their subcellular distribution is affected by polysome-disrupting drugs. Under oxidative stress, both Staufen 1 and Staufen 2 are recruited into stress granules (SGs), which are stress-induced organelles containing transiently silenced messengers. Staufen SGs contain the poly(A)-binding protein (PABP), the RNA binding proteins HuR and TIAR, and small but not large ribosomal subunits. Staufen recruitment into perinuclear SGs is paralleled by a similar change in the overall localization of polyadenylated RNA. Under the same conditions, the distribution of recently transcribed and exported mRNAs is not affected. Our results indicate that Staufen 1 and Staufen 2 are novel and ubiquitous SG components and suggest that Staufen RNPs are involved in repositioning of most polysomal mRNAs, but not of recently synthesized transcripts, during the stress response. PMID- 15525675 TI - Requirement of the Caenorhabditis elegans RapGEF pxf-1 and rap-1 for epithelial integrity. AB - The Rap-pathway has been implicated in various cellular processes but its exact physiological function remains poorly defined. Here we show that the Caenorhabditis elegans homologue of the mammalian guanine nucleotide exchange factors PDZ-GEFs, PXF-1, specifically activates Rap1 and Rap2. Green fluorescent protein (GFP) reporter constructs demonstrate that sites of pxf-1 expression include the hypodermis and gut. Particularly striking is the oscillating expression of pxf-1 in the pharynx during the four larval molts. Deletion of the catalytic domain from pxf-1 leads to hypodermal defects, resulting in lethality. The cuticle secreted by pxf-1 mutants is disorganized and can often not be shed during molting. At later stages, hypodermal degeneration is seen and animals that reach adulthood frequently die with a burst vulva phenotype. Importantly, disruption of rap-1 leads to a similar, but less severe phenotype, which is enhanced by the simultaneous removal of rap-2. In addition, the lethal phenotype of pxf-1 can be rescued by expression of an activated version of rap-1. Together these results demonstrate that the pxf-1/rap pathway in C. elegans is required for maintenance of epithelial integrity, in which it probably functions in polarized secretion. PMID- 15525677 TI - Mitotic regulation of protein 4.1R involves phosphorylation by cdc2 kinase. AB - The nonerythrocyte isoform of the cytoskeletal protein 4.1R (4.1R) is associated with morphologically dynamic structures during cell division and has been implicated in mitotic spindle function. In this study, we define important 4.1R isoforms expressed in interphase and mitotic cells by RT-PCR and mini-cDNA library construction. Moreover, we show that 4.1R is phosphorylated by p34cdc2 kinase on residues Thr60 and Ser679 in a mitosis-specific manner. Phosphorylated 4.1R135 isoform(s) associate with tubulin and Nuclear Mitotic Apparatus protein (NuMA) in intact HeLa cells in vivo as well as with the microtubule-associated proteins in mitotic asters assembled in vitro. Recombinant 4.1R135 is readily phosphorylated in mitotic extracts and reconstitutes mitotic aster assemblies in 4.1R-immunodepleted extracts in vitro. Furthermore, phosphorylation of these residues appears to be essential for the targeting of 4.1R to the spindle poles and for mitotic microtubule aster assembly in vitro. Phosphorylation of 4.1R also enhances its association with NuMA and tubulin. Finally, we used siRNA inhibition to deplete 4.1R from HeLa cells and provide the first direct genetic evidence that 4.1R is required to efficiently focus mitotic spindle poles. Thus, we suggest that 4.1R is a member of the suite of direct cdc2 substrates that are required for the establishment of a bipolar spindle. PMID- 15525676 TI - ERdj3, a stress-inducible endoplasmic reticulum DnaJ homologue, serves as a cofactor for BiP's interactions with unfolded substrates. AB - We recently identified ERdj3 as a component of unassembled immunoglobulin (Ig) heavy chain:BiP complexes. ERdj3 also associates with a number of other protein substrates, including unfolded light chains, a nonsecreted Ig light chain mutant, and the VSV-G ts045 mutant at the nonpermissive temperature. We produced an ERdj3 mutant that was unable to stimulate BiP's ATPase activity in vitro or to bind BiP in vivo. This mutant retained the ability to interact with unfolded protein substrates, suggesting that ERdj3 binds directly to proteins instead of via interactions with BiP. BiP remained bound to unfolded light chains longer than ERdj3, which interacted with unfolded light chains initially, but quickly disassociated before protein folding was completed. This suggests that ERdj3 may bind first to substrates and serve to inhibit protein aggregation until BiP joins the complex, whereas BiP remains bound until folding is complete. Moreover, our findings support a model where interactions with BiP help trigger the release of ERdj3 from the substrate:BiP complex. PMID- 15525678 TI - Coupling of posterior cytoskeletal morphogenesis to the G1/S transition in the Trypanosoma brucei cell cycle. AB - The expression levels of four Cdc2-related kinases (CRK1, 2, 4, and 6) in the procyclic form of Trypanosoma brucei were knocked down in pairs using the RNA interference (RNAi) technique. A double knockdown of CRK1 and CRK2 resulted in arrested cell growth in the G1 phase accompanied by an apparent cessation of nuclear DNA synthesis. The arrested cells became elongated at the posterior end like the G1-phase cells generated by knockdown of CycE1/CYC2 in a previous study. However, approximately 5% of the G1 cells in the current study also possessed multiply branched posterior ends, which have not previously been observed in T. brucei. DAPI and immunofluorescence staining showed a single nucleus, kinetoplast, basal body, and flagellum in the anterior portion of each G1 cell. The split and grossly extended posterior ends were heavily stained with antibodies to tyrosinated alpha-tubulin, suggesting an accumulation of newly synthesized microtubules. A significant population of anucleate cells (zoids), apparently derived from kinetoplast-dictated cytokinesis and cell division of the G1 cells, also had extended and branched posterior ends filled with newly synthesized microtubules. This continued posterior extension of microtubules in the G1 cells and zoids suggests that CRK1 and CRK2 exert a coordinated control on G1/S passage and the limited growth of the microtubule corset toward the posterior end. This connection may provide a new insight into the mechanism of morphological maintenance of an ancient protist during its cell cycle progression. PMID- 15525679 TI - Gap junctions regulate extracellular signal-regulated kinase signaling to affect gene transcription. AB - Osteoblasts are highly coupled by gap junctions formed by connexin43. Overexpression of connexin45 in osteoblasts results in decreased chemical and electrical coupling and reduces gene transcription from connexin response elements (CxREs) in the osteocalcin and collagen Ialpha1 promoters. Here, we demonstrate that transcription from the gap junction-dependent osteocalcin CxRE is regulated by extracellular signal-regulated protein kinase (ERK) and phosphatidylinositol 3-kinase (PI3K) cascades. Overexpression of a constitutively active mitogen-activated protein kinase kinase (MEK), Raf, or Ras can increase transcription more than twofold of the CxRE, whereas inhibition of MEK or PI3K can decrease transcription threefold from the osteocalcin CxRE. Importantly, disruption of gap junctional communication by overexpression of connexin45 or treatment with pharmacological inhibitors of gap junctions results in reduced Raf, ERK, and Akt activation. The consequence of attenuated gap junction dependent signal cascade activation is a decrease in Sp1 phosphorylation by ERK, resulting in decreased Sp1 recruitment to the CxRE and inhibited gene transcription. These data establish that ERK/PI3K signaling is required for the optimal elaboration of transcription from the osteocalcin CxRE, and that disruption of gap junctional communication attenuates the ability of cells to respond to an extracellular cue, presumably by limiting the propagation of second messengers among adjacent cells by connexin43-gap junctions. PMID- 15525680 TI - Mycobacterium tuberculosis functional network analysis by global subcellular protein profiling. AB - Trends in increased tuberculosis infection and a fatality rate of approximately 23% have necessitated the search for alternative biomarkers using newly developed postgenomic approaches. Here we provide a systematic analysis of Mycobacterium tuberculosis (Mtb) by directly profiling its gene products. This analysis combines high-throughput proteomics and computational approaches to elucidate the globally expressed complements of the three subcellular compartments (the cell wall, membrane, and cytosol) of Mtb. We report the identifications of 1044 proteins and their corresponding localizations in these compartments. Genome based computational and metabolic pathways analyses were performed and integrated with proteomics data to reconstruct response networks. From the reconstructed response networks for fatty acid degradation and lipid biosynthesis pathways in Mtb, we identified proteins whose involvements in these pathways were not previously suspected. Furthermore, the subcellular localizations of these expressed proteins provide interesting insights into the compartmentalization of these pathways, which appear to traverse from cell wall to cytoplasm. Results of this large-scale subcellular proteome profile of Mtb have confirmed and validated the computational network hypothesis that functionally related proteins work together in larger organizational structures. PMID- 15525681 TI - R-Ras controls membrane protrusion and cell migration through the spatial regulation of Rac and Rho. AB - Although it is known that the spatial coordination of Rac and Rho activity is essential for cell migration, the molecular mechanisms regulating these GTPases during migration are unknown. We found that the expression of constitutively activated R-Ras (38V) blocked membrane protrusion and random migration. In contrast, expression of dominant negative R-Ras (41A) enhanced migrational persistence and membrane protrusion. Endogenous R-Ras is necessary for cell migration, as cells that were transfected with siRNA for R-Ras did not migrate. Expression of R-Ras (38V) decreased Rac activity and increased Rho activity around the entire cell periphery, whereas expression of dominant negative R-Ras (41A) showed the converse, suggesting that R-Ras can spatially activate Rho and inactivate Rac. Consistent with this role, endogenous R-Ras localized and was preferentially activated at the leading edge of migratory cells in response to adhesion. The effects of R-Ras on cell migration are mediated by PI3-Kinase, as an effector mutant that uncouples PI3-Kinase binding from R-Ras (38V) rescued migration. From these data, we hypothesize that R-Ras plays a key role in cell migration by locally regulating the switch from Rac to Rho activity after membrane protrusion and adhesion. PMID- 15525682 TI - Insulin inhibits platelet-derived growth factor-induced cell proliferation. AB - Cellular behavior can be considered to be the result of a very complex spatial and temporal integration of intracellular and extracellular signals. These signals arise from serum-soluble factors as well as from cell-substrate or cell cell interactions. The current approach in mitogenesis studies is generally to analyze the effect of a single growth factor on serum-starved cells. In this context, a metabolic hormone such as insulin is found to be a mitogenic agent in many cellular types. In the present study, we have considered the effect of insulin stimulation in platelet-derived growth factor (PDGF)-activated NIH-3T3 and C2C12 cells. Our results show that insulin is able to inhibit strongly both NIH-3T3 and C2C12 cell growth induced by PDGF, one of the most powerful mitotic agents for these cell types. This inhibitory effect of insulin is due primarily to a premature down-regulation of the PDGF receptor. Thus, when NIH-3T3 or C2C12 cells are stimulated with both PDGF and insulin, we observe a decrease in PDGF receptor phosphorylation with respect to cells treated with PDGF alone. In particular, we find that costimulation with insulin leads to a reduced production of H2O2 with respect to cell stimulation with PDGF alone. The relative low concentration of H2O2 in PDGF/insulin-costimulated cell leads to a limited down regulation of protein tyrosine phosphatases, and, consequently, to a reduced PDGF receptor phosphorylation efficiency. The latter is very likely to be responsible for the insulin-dependent inhibition of PDGF-receptor mitogenic signaling. PMID- 15525683 TI - Ionic mechanism for contractile response to hyposmotic challenge in canine basilar arteries. AB - A hyposmotic challenge elicited contraction of isolated canine basilar arteries. The contractile response was nearly abolished by the removal of extracellular Ca(2+) and by the voltage-dependent Ca(2+) channel (VDCC) blocker nicardipine, but it was unaffected by thapsigargin, which depletes intracellular Ca(2+) stores. The contraction was also inhibited by Gd(3+) and ruthenium red, cation channel blockers, and Cl(-) channel blockers DIDS and niflumic acid. The reduction of extracellular Cl(-) concentrations enhanced the hypotonically induced contraction. Patch-clamp analysis showed that a hyposmotic challenge activated outwardly rectifying whole cell currents in isolated canine basilar artery myocytes. The reversal potential of the current was shifted toward negative potentials by reductions in intracellular Cl(-) concentration, indicating that the currents were carried by Cl(-). Moreover, the currents were abolished by 10 mM BAPTA in the pipette solution and by the removal of extracellular Ca(2+). Taken together, these results suggest that a hyposmotic challenge activates cation channels, which presumably cause Ca(2+) influx, thereby activating Ca(2+)-activated Cl(-) channels. The subsequent membrane depolarization is likely to increase Ca(2+) influx through VDCC and elicit contraction. PMID- 15525684 TI - Polarized expression of human P2Y receptors in epithelial cells from kidney, lung, and colon. AB - Eight human G protein-coupled P2Y receptors (P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(11), P2Y(12), P2Y(13), and P2Y(14)) that respond to extracellular nucleotides have been molecularly identified and characterized. P2Y receptors are widely expressed in epithelial cells and play an important role in regulating epithelial cell function. Functional studies assessing the capacity of various nucleotides to promote increases in short-circuit current (I(sc)) or Ca(2+) mobilization have suggested that some subtypes of P2Y receptors are polarized with respect to their functional activity, although these results often have been contradictory. To investigate the polarized expression of the family of P2Y receptors, we determined the localization of the entire P2Y family after expression in Madin Darby canine kidney (MDCK) type II cells. Confocal microscopy of polarized monolayers revealed that P2Y(1), P2Y(11), P2Y(12), and P2Y(14) receptors reside at the basolateral membrane, P2Y(2), P2Y(4), and P2Y(6) receptors are expressed at the apical membrane, and the P2Y(13) receptor is unsorted. Biotinylation studies and I(sc) measurements in response to the appropriate agonists were consistent with the polarized expression observed in confocal microscopy. Expression of the G(q)-coupled P2Y receptors (P2Y(1), P2Y(2), P2Y(4), P2Y(6), and P2Y(11)) in lung and colonic epithelial cells (16HBE14o- and Caco-2 cells, respectively) revealed a targeting profile nearly identical to that observed in MDCK cells, suggesting that polarized targeting of these P2Y receptor subtypes is not a function of the type of epithelial cell in which they are expressed. These experiments highlight the highly polarized expression of P2Y receptors in epithelial cells. PMID- 15525685 TI - Cellular mechanisms involved in CO(2) and acid signaling in chemosensitive neurons. AB - An increase in CO(2)/H(+) is a major stimulus for increased ventilation and is sensed by specialized brain stem neurons called central chemosensitive neurons. These neurons appear to be spread among numerous brain stem regions, and neurons from different regions have different levels of chemosensitivity. Early studies implicated changes of pH as playing a role in chemosensitive signaling, most likely by inhibiting a K(+) channel, depolarizing chemosensitive neurons, and thereby increasing their firing rate. Considerable progress has been made over the past decade in understanding the cellular mechanisms of chemosensitive signaling using reduced preparations. Recent evidence has pointed to an important role of changes of intracellular pH in the response of central chemosensitive neurons to increased CO(2)/H(+) levels. The signaling mechanisms for chemosensitivity may also involve changes of extracellular pH, intracellular Ca(2+), gap junctions, oxidative stress, glial cells, bicarbonate, CO(2), and neurotransmitters. The normal target for these signals is generally believed to be a K(+) channel, although it is likely that many K(+) channels as well as Ca(2+) channels are involved as targets of chemosensitive signals. The results of studies of cellular signaling in central chemosensitive neurons are compared with results in other CO(2)- and/or H(+)-sensitive cells, including peripheral chemoreceptors (carotid body glomus cells), invertebrate central chemoreceptors, avian intrapulmonary chemoreceptors, acid-sensitive taste receptor cells on the tongue, and pain-sensitive nociceptors. A multiple factors model is proposed for central chemosensitive neurons in which multiple signals that affect multiple ion channel targets result in the final neuronal response to changes in CO(2)/H(+). PMID- 15525686 TI - Probing the iron pool. Focus on "Detection of intracellular iron by its regulatory effect". PMID- 15525687 TI - Surfing the lipid bilayer: how basolateral insulin receptors regulate Na(+) channels in the apical cell membrane. Focus on "Real-time three-dimensional imaging of lipid signal transduction: apical membrane insertion of epithelial Na(+) channels". PMID- 15525688 TI - Activation of PLC-delta1 by Gi/o-coupled receptor agonists. AB - The mechanism of phospholipase (PLC)-delta activation by G protein-coupled receptor agonists was examined in rabbit gastric smooth muscle. Ca(2+) stimulated an eightfold increase in PLC-delta1 activity in permeabilized muscle cells. Treatment of dispersed or cultured muscle cells with three G(i/o)-coupled receptor agonists (somatostatin, delta-opioid agonist [D-Pen(2),D Pen(5)]enkephalin, and A(1) agonist cyclopentyl adenosine) caused delayed increase in phosphoinositide (PI) hydrolysis (8- to 10-fold) that was strongly inhibited by overexpression of dominant-negative PLC-delta1(E341R/D343R; 65-76%) or constitutively active RhoA(G14V). The response coincided with capacitative Ca(2+) influx and was not observed in the absence of extracellular Ca(2+), but was partly inhibited by nifedipine (16-30%) and strongly inhibited by SKF-96365, a blocker of store-operated Ca(2+) channels. Treatment of the cells with a G(q/13)-coupled receptor agonist, CCK-8, caused only transient, PLC-beta1 mediated PI hydrolysis. Unlike G(i/o)-coupled receptor agonists, CCK-8 activated RhoA and stimulated RhoA:PLC-delta1 association. Inhibition of RhoA activity with C3 exoenzyme or by overexpression of dominant-negative RhoA(T19N) or Galpha(13) minigene unmasked a delayed increase in PI hydrolysis that was strongly inhibited by coexpression of PLC-delta1(E341R/D343R) or by SKF-96365. Agonist-independent capacitative Ca(2+) influx induced by thapsigargin stimulated PI hydrolysis (8 fold), which was partly inhibited by nifedipine ( approximately 25%) and strongly inhibited by SKF-96365 ( approximately 75%) and in cells expressing PLC delta1(E341R/D343R). Agonist-independent Ca(2+) release or Ca(2+) influx via voltage-gated Ca(2+) channels stimulated only moderate PI hydrolysis (2- to 3 fold), which was abolished by PLC-delta1 antibody or nifedipine. We conclude that PLC-delta1 is activated by G(i/o)-coupled receptor agonists that do not activate RhoA. The activation is preferentially mediated by Ca(2+) influx via store operated Ca(2+) channels. PMID- 15525689 TI - Calmodulin reverses rundown of L-type Ca(2+) channels in guinea pig ventricular myocytes. AB - Calmodulin (CaM) is implicated in regulation of Ca(2+) channels as a Ca(2+) sensor. The effect of CaM on rundown of L-type Ca(2+) channels in inside-out patch form was investigated in guinea pig ventricular myocytes. Ca(2+) channel activity disappeared within 1-3 min and did not reappear when the patch was excised and exposed to an artificial intracellular solution. However, application of CaM (0.03, 0.3, 3 microM) + 3 mM ATP to the intracellular solution within 1 min after patch excision resulted in dose-dependent activation of channel activity. Channel activity averaged 11.2%, 94.7%, and 292.9%, respectively, of that in cell-attached mode. Channel activity in inside-out patch mode was induced by CaM + ATP at nanomolar Ca(2+) concentrations ([Ca(2+)]); however, increase to micromolar [Ca(2+)] rapidly inactivated the channel activity induced, revealing that the effect of CaM on the channel was Ca(2+) dependent. At the 2nd, 4th, 6th, 8th, and 10th minutes after patch excision, CaM (0.75 microM) + ATP induced Ca(2+) channel activity to 150%, 100%, 96.9%, 29.3%, and 16.6%, respectively, revealing a time-dependent action of CaM on the channel. CaM added with adenosine 5'-(beta,gamma-imido)triphosphate (AMP-PNP) also induced channel activity, although with much lower potency and shorter duration. Protein kinase inhibitors KN-62, CaM-dependent protein kinase (CaMK)II 281-309, autocamtide-related CaMKII inhibitor peptide, and K252a (each 1-10 microM) did not block the effect of CaM, indicating that the effect of CaM on the Ca(2+) channel was phosphorylation independent. Neither CaM nor ATP alone induced Ca(2+) channel activity, showing a cooperative effect of CaM and ATP on the Ca(2+) channel. These results suggest that CaM is a crucial regulatory factor of Ca(2+) channel basal activity. PMID- 15525690 TI - Stabilization of Nrf2 by tBHQ confers protection against oxidative stress-induced cell death in human neural stem cells. AB - Recent studies indicate that NF-E2 related factor 2 (Nrf2) is a substrate for the ubiquitin-proteasome pathway. The present study is aimed to determine whether increased protein stability is a mechanism by which quinone compounds, like tert butylhydroquinone (tBHQ), may enhance Nrf2-mediated transcriptional activation and subsequent antioxidant protection. H2O2-induced necrotic cell death, evidenced by transmission electronic microscope (TEM) imaging with no caspase 3 activation and PARP cleavage, was significantly attenuated by pretreatment with tBHQ or overexpression of Nrf2 through advenovirus-mediated infection in human neural stem cells (hNSCs). Microarray analysis showed that those identified antioxidant genes, responsible for antiapoptotic action in IMR-32 cells (J. Li et al., 2002, J. Biol. Chem. 277, 388-394), were also coordinately upregulated through Nrf2-dependent antioxidant responsive element (ARE) activation in hNSC. The stabilization of Nrf2 by tBHQ in IMR-32 cells was evidenced by a pulse-chase assay showing no significant increase in Nrf2 protein synthesis after tBHQ treatment, and by ubiquitin immunoprecipitation showing that tBHQ stabilized ubiquitinated Nrf2. An in vitro proteasomal activity assay showed that tBHQ did not act as a 20S/26S proteasome inhibitor. Nrf2 stabilization by tBHQ also was observed in hNSCs. Taken together, this study suggests that identified antioxidant genes, which were upregulated through tBHQ induced Nrf2 stabilization, confer protection on target cells against H2O2-induced apoptotic cell death in neuroblastoma cells as well as the necrotic cell death in the hNSC. Nrf2 stabilization by pharmacological modulation or adenovirus-mediated Nrf2 overexpression, therefore, might be viable strategies to prevent a wide-spectrum of oxidative stress-related neuronal cell injuries. PMID- 15525691 TI - 2,2',4,6,6'-Pentachlorobiphenyl-induced apoptosis is limited by cyclooxygenase-2 induction. AB - Polychlorinated biphenyls (PCBs), a group of persistent and widespread environmental pollutants, are considered to be immunotoxic, carcinogenic, and to induce apoptosis. However, the cellular mechanisms underlying the action of PCBs have not been established. Here, we investigated the effects of PCBs on the induction of cyclooxygenase-2 (COX-2). Among the several congeners examined, only 2,2',4,6,6'-pentachlorobiphenyl (PeCB) specifically increased the COX-2 promoter activity, and the levels of COX-2 mRNA and protein, and thereby enhanced prostaglandin E2 (PGE2) synthesis in Rat-1 cells. By conducting mutation analyses of the COX-2 promoter and its transcription factor, we found that the CRE site in COX-2 promoter and c-Jun are important for increased COX-2 promoter activity induced by 2,2',4,6,6'-PeCB. In addition, 2,2',4,6,6'-PeCB-stimulated COX-2 induction was reduced by the specific MAPK kinase (MEK) inhibitor, PD98059, and in p53-deficient cells, implying that COX-2 induction requires the activation of ERK1/2 MAPK and p53. The selective COX-2 inhibitor, NS-398, potentiated the 2,2',4,6,6'-PeCB-induced mitochondrial apoptotic pathway involved in Bcl-xL attenuation, cytochrome c release and the subsequent activation of caspase-3. Furthermore, the cell death was prevented by PGE2 treatment, suggesting that 2,2',4,6,6'-PeCB-induced apoptosis is restricted by prostaglandin upregulation by COX-2. Taken together, these results demonstrate that 2,2',4,6,6'-PeCB-induced COX-2 expression may be an important compensatory mechanism for abating 2,2',4,6,6'-PeCB toxicity. PMID- 15525692 TI - Co-culture of primary human mammary fibroblasts and MCF-7 cells as an in vitro breast cancer model. AB - Approximately 60% of all breast tumors are estrogen-responsive and chemicals that show estrogenic or anti-estrogenic properties are able to interact with breast tumor growth. In a breast tumor, adipose stromal cells (fibroblasts) surrounding the epithelial tumor contain the aromatase enzyme, which converts androgens into estrogens. Exposure to aromatase inducers can therefore lead to increased estrogen levels and possibly to accelerated breast tumor growth. Subsequently, breast tumor cells synthesize and secrete elevated levels of factors such as prostaglandin E2 (PGE2), interleukin-6 (IL-6), and IL-6 soluble receptor (IL 6sR), which in turn have the ability to stimulate aromatase gene transcription in fibroblasts, establishing a positive feedback loop. In this study, a technique that allows for culturing MCF-7 epithelial breast tumor cells and healthy primary human mammary fibroblasts together in one compartment was developed. To establish the positive feedback loop, the co-culture was exposed to estrogenic compounds. RNA was isolated and reverse-transcriptase polymerase chain reaction (RT-PCR) was performed on the aromatase and pS2 genes. Exposure of the co-culture to estradiol (E2), diethylstilbestrol (DES), and bisphenol-A (BPA), resulted in a three- to seven-fold increase of pS2 transcription levels. Furthermore, pS2 transcription levels increased even more when the aromatase substrate testosterone (20 nM) was present in the co-culture medium. Exposure of the co-culture to the aromatase inducer dexamethasone (DEX) resulted in increased pS2 transcription levels, as well as increased aromatase transcription levels. Simultaneous exposure to DEX and the synthetic anti-estrogen ICI 182,780 almost completely blocked the pS2 response. The aromatase induction response was not altered by ICI 182,780 treatment. Simultaneous exposure to DEX and the non-steroidal aromatase inhibitor fadrozole, abolished the effect of the presence of testosterone in the co-culture medium, but did not result in pS2 gene transcription levels as low as seen after exposure to ICI 182,780. These observations indicate the presence of a positive feedback loop in our co-culture system. This co-culture provides a more sophisticated and sensitive system to detect direct and indirect estrogenic effects of compounds and their possible effects on breast tumor promotion. PMID- 15525693 TI - Methoxychlor induces proliferation of the mouse ovarian surface epithelium. AB - While the pesticide methoxychlor (MXC) has a variety of adverse effects on the female reproductive system, the effects of MXC on the ovarian surface epithelium (OSE) are unknown. Thus, this study tested the hypothesis that MXC alters the growth of the OSE. Mouse OSE cells were isolated by enzymatic digestion and cultured with vehicle, 3 microM of MXC, or 3 microM of 2,2-bis[p-hydroxyphenyl] 1,1,1,-trichloroethane (HPTE) for 14 days. After culture, proliferation and apoptosis were assessed by measurement of cell density, immunohistochemistry, and real-time polymerase chain reaction. Cell density was 66% greater for MXC-treated cells and 95% greater for HPTE-treated cells than controls (p < or = 0.05). The estrogen receptor blocker ICI 182,780 abolished MXC- and HPTE-induced increases in cell density. Proliferating cell nuclear antigen (PCNA) staining was positive in only 22 +/- 2.3% of controls, compared to 35 +/- 2.4% of MXC-treated cells and 40 +/- 2.4% of HPTE-treated cells (p < or = 0.05). The cell cycle regulators, cyclinD2 and cdk4, were significantly increased in MXC- and HPTE-treated cells compared to controls. The ApopTag assay demonstrated apoptotic cells in 4.8 +/- 0.45% of controls, 2.2 +/- 0.56% of MXC-treated cells, and 2.1 +/- 0.33% of HPTE treated cells (p < or = 0.005). Expression of bcl-2 was significantly increased in MXC- and HPTE-treated cells, while bax was decreased in MXC- and HPTE-treated cells compared to controls. Collectively, these data indicate that MXC and HPTE stimulate OSE cell growth by increasing proliferation and inhibiting apoptosis. Further, since ICI 182,780 blocked MXC- and HPTE-induced OSE growth, these data suggest that the effects of MXC and HPTE on the OSE are mediated by estrogen receptors. PMID- 15525694 TI - Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry. AB - The classical laboratory tests for exposure to organophosphorus toxicants (OP) are inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in blood. In a search for new biomarkers of OP exposure, we treated mice with a biotinylated organophosphorus agent, FP-biotin. The biotinylated proteins in muscle were purified by binding to avidin-Sepharose, separated by gel electrophoresis, digested with trypsin, and identified from their fragmentation patterns on a quadrupole time-of-flight mass spectrometer. Albumin and ES1 carboxylesterase (EC 3.1.1.1) were found to be major targets of FP-biotin. These FP-biotinylated proteins were also identified in mouse plasma by comparing band patterns on nondenaturing gels stained for albumin and carboxylesterase activity, with band patterns on blots hybridized with Streptavidin Alexa-680. Two additional FP-biotin targets, AChE (EC 3.1.1.7) and BChE (EC 3.1.1.8), were identified in mouse plasma by finding that enzyme activity was inhibited 50-80%. Mouse plasma contained eight additional FP-biotinylated bands whose identity has not yet been determined. In vitro experiments with human plasma showed that chlorpyrifos oxon, echothiophate, malaoxon, paraoxon, methyl paraoxon, diazoxon, diisopropylfluorophosphate, and dichlorvos competed with FP-biotin for binding to human albumin. Though experiments with purified albumin have previously shown that albumin covalently binds OP, this is the first report of OP binding to albumin in a living animal. Carboxylesterase is not a biomarker in man because humans have no carboxylesterase in blood. It is concluded that OP bound to albumin could serve as a new biomarker of OP exposure in man. PMID- 15525695 TI - Chaperone proteins involved in troglitazone-induced toxicity in human hepatoma cell lines. AB - Troglitazone (TRO), an effective thiazolidinedione antidiabetic agent, was reported to produce idiosyncratic hepatotoxic effects in some individuals. In contrast, rosiglitazone (RSG), in the same group of agents, has no significant toxic effects and now is widely used. In this study, human hepatoma (HepG2) cell lines were exposed to various doses of TRO as well as RSG (0, 25, 50, and 75 microM) for 48 h. Cell lysates were separated by two-dimensional electrophoresis, and the gels were stained with coomassie brilliant blue to compare the spot profiles. The greatest protein expression at a MW of 75 kDa and isoelectric point of 5 was specifically increased with TRO treatments of 50 and 75 microM. The spot was identified as a mixture of immunoglobulin heavy chain binding protein (BiP) and, to a lesser extent, protein disulfide isomerase-related protein (PDIrp). Immunoblot analyses showed that the BiP protein was dose-dependently increased by TRO treatment and, to a lower degree, by RSG. These effects were also correlated with the high induction of BiP mRNA by TRO (50 and 75 microM) and the lower induction by RSG. However, both treatments showed no significant effects on PDIrp expression. The toxic effects of TRO in relation to the overexpression of BiP were also demonstrated in HLE cells, another human hepatoma cell line. In HLE cells, the inhibition of BiP expression by small interference RNA rendered cells more susceptible to the toxic effects of TRO. These results suggest that the overexpression of BiP is a defense mechanism of the endoplasmic reticulum in response to TRO-induced toxicity. PMID- 15525696 TI - The comparative method rules! Codon volatility cannot detect positive Darwinian selection using a single genome sequence. AB - All established methods for detecting positive selection at the molecular level rely on comparisons between nucleotide sequences. An exceptional method that purports to detect selection on the basis of a single genomic sequence has recently been proposed. This method uses a measure called "codon volatility," defined for each codon as the ratio between the number of nonsynonymous codons that differ from the codon under study at a single nucleotide position and the number of sense codons that differ from the codon under study at a single nucleotide position. Here, we examine various properties of codon volatility and its derivatives and use simulation of evolutionary processes to determine whether they can be used to detect selective pressures. Codons for only four amino acids (glycine, leucine, arginine, and serine) show any variation in codon volatility. Thus, codon volatility is mainly a proxy for amino acid usage, rather than for codon usage, with 65% of all synonymous changes and 27% of all nonsynonymous changes being undetectable by this measure. Genes identified by the volatility method as being subject to positive selection tend to have idiosyncratic amino acid compositions (e.g., they are glycine rich or arginine poor). An additional property of codon volatility is the near zero variance of its mean expectation, which translates into overestimated statistical significance estimates, especially in the absence of corrections for multiple comparisons. A comparison with measures of selection inferred through comparative methodology reveals no relationship between the results of the two methods. Finally, we show that codon volatility can increase in the absence of positive Darwinian selection; that is, increased codon volatility is not indicative of positive selection. PMID- 15525697 TI - A bayesian analysis of metazoan mitochondrial genome arrangements. AB - Genome arrangements are a potentially powerful source of information to infer evolutionary relationships among distantly related taxa. Mitochondrial genome arrangements may be especially informative about metazoan evolutionary relationships because (1) nearly all animals have the same set of definitively homologous mitochondrial genes, (2) mitochondrial genome rearrangement events are rare relative to changes in sequences, and (3) the number of possible mitochondrial genome arrangements is huge, making convergent evolution of genome arrangements appear highly unlikely. In previous studies, phylogenetic evidence in genome arrangement data is nearly always used in a qualitative fashion-the support in favor of clades with similar or identical genome arrangements is considered to be quite strong, but is not quantified. The purpose of this article is to quantify the uncertainty among the relationships of metazoan phyla on the basis of mitochondrial genome arrangements while incorporating prior knowledge of the monophyly of various groups from other sources. The work we present here differs from our previous work in the statistics literature in that (1) we incorporate prior information on classifications of metazoans at the phylum level, (2) we describe several advances in our computational approach, and (3) we analyze a much larger data set (87 taxa) that consists of each unique, complete mitochondrial genome arrangement with a full complement of 37 genes that were present in the NCBI (National Center for Biotechnology Information) database at a recent date. In addition, we analyze a subset of 28 of these 87 taxa for which the non-tRNA mitochondrial genomes are unique where the assumption of our inversion-only model of rearrangement is more plausible. We present summaries of Bayesian posterior distributions of tree topology on the basis of these two data sets. PMID- 15525698 TI - Codon volatility as an indicator of positive selection: data from eukaryotic genome comparisons. AB - It has been suggested that codon volatility (the proportion of the point-mutation neighbors of a codon that encode different amino acids) can be used as an index of past positive selection. We compared codon volatility with patterns of synonymous and nonsynonymous nucleotide substitution in genome-wide comparisons of orthologous genes between three pairs of related genomes: (1) the protists Plasmodium falciparum and P. yoelii, (2) the fungi Saccharomyces cerevisiae and S. paradoxus, and (3) the mammals mouse and rat. Codon volatility was not consistently associated with an elevated rate of nonsynonymous substitution, as would be expected under positive selection. Rather, the most consistent and powerful correlate of elevated codon volatility was nucleotide content at the second codon position, as expected, given the nature of the genetic code. PMID- 15525699 TI - Positive selection in the carbohydrate recognition domains of sea urchin sperm receptor for egg jelly (suREJ) proteins. AB - A wealth of evidence shows that protein-carbohydrate recognition mediates the steps of gamete interaction during fertilization. Carbohydrate-recognition domains (CRDs) comprise a large family of ancient protein modules of approximately 120 amino acids, having the same protein fold, that bind terminal sugar residues on glycoproteins and polysaccharides. Sea urchin sperm express three suREJ (sea urchin receptor for egg jelly) proteins on their plasma membranes. suREJ1 has two CRDs, whereas suREJ2 and suREJ3 both have one CRD. suREJ1 binds the fucose sulfate polymer (FSP) of egg jelly to induce the sperm acrosome reaction. The structure of FSP is species specific. Therefore, the suREJ1 CRDs could encode molecular recognition between sperm and egg underlying the species-specific induction of the acrosome reaction. The functions of suREJ2 and suREJ3 have not been explored, but suREJ3 is exclusively localized on the plasma membrane over the sperm acrosomal vesicle and is physically associated with sea urchin polycystin-2, a known cation channel. An evolutionary analysis of these four CRDs was performed for six sea urchin species. Phylogenetic analysis shows that these CRDs were already differentiated in the common ancestor of these six sea urchins. The CRD phylogeny agrees with previous work on these species based on one nuclear gene and several mitochondrial genes. Maximum likelihood shows that positive selection acts on these four CRDs. Threading the suREJ CRDs onto the prototypic CRD crystal structure shows that many of the sites under positive selection are on extended loops, which are involved in saccharide binding. This is the first demonstration of positive selection in CRDs and is another example of positive selection acting on the evolution of gamete recognition proteins. PMID- 15525700 TI - Nonhomogeneous model of sequence evolution indicates independent origins of primary endosymbionts within the enterobacteriales (gamma-Proteobacteria). AB - Standard methods of phylogenetic reconstruction are based on models that assume homogeneity of nucleotide composition among taxa. However, this assumption is often violated in biological data sets. In this study, we examine possible effects of nucleotide heterogeneity among lineages on the phylogenetic reconstruction of a bacterial group that spans a wide range of genomic nucleotide contents: obligately endosymbiotic bacteria and free-living or commensal species in the gamma-Proteobacteria. We focus on AT-rich primary endosymbionts to better understand the origins of obligately intracellular lifestyles. Previous phylogenetic analyses of this bacterial group point to the importance of accounting for base compositional variation in estimating relationships, particularly between endosymbiotic and free-living taxa. Here, we develop an approach to compare susceptibility of various phylogenetic reconstruction methods to the effects of nucleotide heterogeneity. First, we identify candidate trees of gamma-Proteobacteria groEL and 16S rRNA using approaches that assume homogeneous and stationary base composition, including Bayesian, maximum likelihood, parsimony, and distance methods. We then create permutations of the resulting candidate trees by varying the placement of the AT-rich endosymbiont Buchnera. These permutations are evaluated under the nonhomogeneous and nonstationary maximum likelihood model of Galtier and Gouy, which allows equilibrium base content to vary among examined lineages. Our results show that commonly used phylogenetic methods produce incongruent trees of the Enterobacteriales, and that the placement of Buchnera is especially unstable. However, under a nonhomogeneous model, various groEL and 16S rRNA phylogenies that separate Buchnera from other AT-rich endosymbionts (Blochmannia and Wigglesworthia) have consistently and significantly higher likelihood scores. Blochmannia and Wigglesworthia appear to have evolved from secondary endosymbionts, and represent an origin of primary endosymbiosis that is independent from Buchnera. This application of a nonhomogeneous model offers a computationally feasible way to test specific phylogenetic hypotheses for taxa with heterogeneous and nonstationary base composition. PMID- 15525701 TI - Molecular population genetics and the search for adaptive evolution in plants. AB - The first papers on plant molecular population genetics were published approximately 10 years ago. Since that time, well over 50 additional studies of plant nucleotide polymorphism have been published, and many of these studies focused on detecting the signature of balancing or positive selection at a locus. In this review, we discuss some of the theoretical and statistical issues surrounding the detection of selection, with focus on plant populations, and we also summarize the empirical plant molecular population genetics literature. At face value, the literature suggests that a history of balancing or positive selection in plant genes is rampant. In two well-studied taxa (maize and Arabidopsis) over 20% of studied genes have been interpreted as containing the signature of selection. We argue that this is probably an overstatement of the prevalence of natural selection in plant genomes, for two reasons. First, demographic effects are difficult to incorporate and have generally not been well integrated into the plant population genetics literature. Second, the genes studied to date are not a random sample, so selected genes may be overrepresented. The next generation of studies in plant molecular population genetics requires additional sampling of local populations, explicit comparisons among loci, and improved theoretical methods to control for demography. Eventually, candidate loci should be confirmed by explicit consideration of phenotypic effects. PMID- 15525702 TI - Divergence pattern of duplicate genes in protein-protein interactions follows the power law. AB - The impact of the biological network structures on the divergence between the two copies of one duplicate gene pair involved in the networks has not been documented on a genome scale. Having analyzed the most recently updated Database of Interacting Proteins (DIP) by incorporating the information for duplicate genes of the same age in yeast, we find that there was a highly significantly positive correlation between the level of connectivity of ancient genes and the number of shared partners of their duplicates in the protein-protein interaction networks. This suggests that duplicate genes with a low ancestral connectivity tend to provide raw materials for functional novelty, whereas those duplicate genes with a high ancestral connectivity tend to create functional redundancy for a genome during the same evolutionary period. Moreover, the difference in the number of partners between two copies of a duplicate pair was found to follow a power-law distribution. This suggests that loss and gain of interacting partners for most duplicate genes with a lower level of ancestral connectivity is largely symmetrical, whereas the "hub duplicate genes" with a higher level of ancient connectivity display an asymmetrical divergence pattern in protein-protein interactions. Thus, it is clear that the protein-protein interaction network structures affect the divergence pattern of duplicate genes. Our findings also provide insights into the origin and development of biological networks. PMID- 15525703 TI - The 15-K neutron structure of saccharide-free concanavalin A. AB - The positions of the ordered hydrogen isotopes of a protein and its bound solvent can be determined by using neutron crystallography. Furthermore, by collecting neutron data at cryo temperatures, the dynamic disorder within a protein crystal is reduced, which may lead to improved definition of the nuclear density. It has proved possible to cryo-cool very large Con A protein crystals (>1.5 mm3) suitable for high-resolution neutron and x-ray structure analysis. We can thereby report the neutron crystal structure of the saccharide-free form of Con A and its bound water, including 167 intact D2O molecules and 60 oxygen atoms at 15 K to 2.5-A resolution, along with the 1.65-A x-ray structure of an identical crystal at 100 K. Comparison with the 293-K neutron structure shows that the bound water molecules are better ordered and have lower average B factors than those at room temperature. Overall, twice as many bound waters (as D2O) are identified at 15 K than at 293 K. We note that alteration of bound water orientations occurs between 293 and 15 K; such changes, as illustrated here with this example, could be important more generally in protein crystal structure analysis and ligand design. Methodologically, this successful neutron cryo protein structure refinement opens up categories of neutron protein crystallography, including freeze-trapped structures and cryo to room temperature comparisons. PMID- 15525704 TI - Formation of tabular single-domain magnetite induced by Geobacter metallireducens GS-15. AB - Distinct morphological characteristics of magnetite formed intracellularly by magnetic bacteria (magnetosome) are invoked as compelling evidence for biological activity on Earth and possibly on Mars. Crystals of magnetite produced extracellularly by a variety of bacteria including Geobacter metallireducens GS 15, thermophilic bacteria, and psychrotolerant bacteria are, however, traditionally not thought to have nearly as distinct morphologies. The size and shape of extracellular magnetite depend on the culture conditions and type of bacteria. Under typical CO(2)-rich culture conditions, GS-15 is known to produce superparamagnetic magnetite (crystal diameters of approximately <30 nm). In the current study, we were able to produce a unique form of tabular, single-domain magnetite under nontraditional (low-CO(2)) culture conditions. This magnetite has a distinct crystal habit and magnetic properties. This magnetite could be used as a biosignature to recognize ancient biological activities in terrestrial and extraterrestrial environments and also may be a major carrier of the magnetization in natural sediments. PMID- 15525705 TI - Archaea recruited D-Tyr-tRNATyr deacylase for editing in Thr-tRNA synthetase. AB - Aminoacyl-tRNA synthetases (AARSs) are key players in the maintenance of the genetic code through correct pairing of amino acids with their cognate tRNA molecules. To this end, some AARSs, as well as seeking to recognize the correct amino acid during synthesis of aminoacyl-tRNA, enhance specificity through recognition of mischarged aminoacyl-tRNA molecules in a separate editing reaction. Recently, an editing domain, of uncertain provenance, idiosyncratic to some archaeal ThrRSs has been characterized. Here, sequence analyses and molecular modeling are reported that clearly show a relationship of the archaea specific ThrRS editing domains with d-Tyr-tRNATyr deacylases (DTDs). The model enables the identification of the catalytic site and other substrate binding residues, as well as the proposal of a likely catalytic mechanism. Interestingly, typical DTD sequences, common in bacteria and eukaryotes, are entirely absent in archaea, consistent with an evolutionary scheme in which DTD was co-opted to serve as a ThrRS editing domain in archaea soon after their divergence from eukaryotes. A group of present-day archaebacteria contain a ThrRS obtained from a bacterium by horizontal gene transfer. In some of these cases a vestigial version of the original archaeal ThrRS, of potentially novel function, is maintained. PMID- 15525706 TI - RNA chaperone activity of large ribosomal subunit proteins from Escherichia coli. AB - The ribosome is a highly dynamic ribonucleoprotein machine. During assembly and during translation the ribosomal RNAs must routinely be prevented from falling into kinetic folding traps. Stable occupation of these trapped states may be prevented by proteins with RNA chaperone activity. Here, ribosomal proteins from the large (50S) ribosome subunit of Escherichia coli were tested for RNA chaperone activity in an in vitro trans splicing assay. Nearly a third of the 34 large ribosomal subunit proteins displayed RNA chaperone activity. We discuss a possible role of this function during ribosome assembly and during translation. PMID- 15525707 TI - Assembly of the 30S ribosomal subunit: positioning ribosomal protein S13 in the S7 assembly branch. AB - Studies of Escherichia coli 30S ribosomal subunit assembly have revealed a hierarchical and cooperative association of ribosomal proteins with 16S ribosomal RNA; these results have been used to compile an in vitro 30S subunit assembly map. In single protein addition and omission studies, ribosomal protein S13 was shown to be dependent on the prior association of ribosomal protein S20 for binding to the ribonucleoprotein particle. While the overwhelming majority of interactions revealed in the assembly map are consistent with additional data, the dependency of S13 on S20 is not. Structural studies position S13 in the head of the 30S subunit > 100 A away from S20, which resides near the bottom of the body of the 30S subunit. All of the proteins that reside in the head of the 30S subunit, except S13, have been shown to be part of the S7 assembly branch, that is, they all depend on S7 for association with the assembling 30S subunit. Given these observations, the assembly requirements for S13 were investigated using base-specific chemical footprinting and primer extension analysis. These studies reveal that S13 can bind to 16S rRNA in the presence of S7, but not S20. Additionally, interaction between S13 and other members of the S7 assembly branch have been observed. These results link S13 to the 3' major domain family of proteins, and the S7 assembly branch, placing S13 in a new location in the 30S subunit assembly map where its position is in accordance with much biochemical and structural data. PMID- 15525708 TI - Human microRNAs are processed from capped, polyadenylated transcripts that can also function as mRNAs. AB - The factors regulating the expression of microRNAs (miRNAs), a ubiquitous family of approximately 22-nt noncoding regulatory RNAs, remain undefined. However, it is known that miRNAs are first transcribed as a largely unstructured precursor, termed a primary miRNA (pri-miRNA), which is sequentially processed in the nucleus, to give the approximately 65-nt pre-miRNA hairpin intermediate, and then in the cytoplasm, to give the mature miRNA. Here we have sought to identify the RNA polymerase responsible for miRNA transcription and to define the structure of a full-length human miRNA. We show that the pri-miRNA precursors for nine human miRNAs are both capped and polyadenylated and report the sequence of the full length, approximately 3433-nt pri-miR-21 RNA. This pri-miR-21 gene sequence is flanked 5' by a promoter element able to transcribe heterologous mRNAs and 3' by a consensus polyadenylation sequence. Nuclear processing of pri-miRNAs was found to be efficient, thus largely preventing the nuclear export of full-length pri miRNAs. Nevertheless, an intact miRNA stem-loop precursor located in the 3' UTR of a protein coding gene only moderately inhibited expression of the linked open reading frame, probably because the 3' truncated mRNA could still be exported and expressed. Together, these data show that human pri-miRNAs are not only structurally similar to mRNAs but can, in fact, function both as pri-miRNAs and mRNAs. PMID- 15525709 TI - A computational and experimental approach toward a priori identification of alternatively spliced exons. AB - Alternative splicing is a powerful means of regulating gene expression and enhancing protein diversity. In fact, the majority of metazoan genes encode pre mRNAs that are alternatively spliced to produce anywhere from two to tens of thousands of mRNA isoforms. Thus, an important part of determining the complete proteome of an organism is developing a catalog of all mRNA isoforms. Alternatively spliced exons are typically identified by aligning EST clusters to reference mRNAs or genomic DNA. However, this approach is not useful for genomes that lack robust EST coverage, and tools that enable accurate prediction of alternatively spliced exons would be extraordinarily useful. Here, we use comparative genomics to identify, and experimentally verify, potential alternative exons based solely on their high degree of conservation between Drosophila melanogaster and D. pseudoobscura. At least 40% of the exons that fit our prediction criteria are in fact alternatively spliced. Thus, comparative genomics can be used to accurately predict certain classes of alternative exons without relying on EST data. PMID- 15525710 TI - The RNA catabolic enzymes Rex4p, Rnt1p, and Dbr1p show genetic interaction with trans-acting factors involved in processing of ITS1 in Saccharomyces cerevisiae pre-rRNA. AB - Eukaryotes have two types of ribosomes containing either 5.8SL or 5.8SS rRNA that are produced by alternative pre-rRNA processing. The exact processing pathway for the minor 5.8SL rRNA species is poorly documented. We have previously shown that the trans-acting factor Rrp5p and the RNA exonuclease Rex4p genetically interact to influence the ratio between the two forms of 5.8S rRNA in the yeast Saccharomyces cerevisiae. Here we report a further analysis of ITS1 processing in various yeast mutants that reveals genetic interactions between, on the one hand, Rrp5p and RNase MRP, the endonuclease required for 5.8SS rRNA synthesis, and, on the other, Rex4p, the RNase III homolog Rnt1p, and the debranching enzyme Dbr1p. Yeast cells carrying a temperature-sensitive mutation in RNase MRP (rrp2-1) exhibit a pre-rRNA processing phenotype very similar to that of the previously studied rrp5-33 mutant: ITS2 processing precedes ITS1 processing, 5.8SL rRNA becomes the major species, and ITS1 is processed at the recently reported novel site A4 located midway between sites A2 and A3. As in the rrp5-Delta3 mutant, all of these phenotypical processing features disappear upon inactivation of the REX4 gene. Moreover, inactivation of the DBR1 gene in rrp2-1, or the RNT1 gene in rrp5 Delta3 mutant cells also negates the effects of the original mutation on pre-rRNA processing. These data link a total of three RNA catabolic enzymes, Rex4p, Rnt1p, and Dbr1p, to ITS1 processing and the relative production of 5.8SS and 5.8SL rRNA. A possible model for the indirect involvement of the three enzymes in yeast pre-rRNA processing is discussed. PMID- 15525711 TI - Generation of a catalytic module on a self-folding RNA. AB - It is theoretically possible to obtain a catalytic site of an artificial ribozyme from a random sequence consisting of a limited numbers of nucleotides. However, this strategy has been inadequately explored. Here, we report an in vitro selection technique that exploits modular construction of a structurally constrained RNA to acquire a catalytic site for RNA ligation from a short random sequence. To practice the selection, a sequence of 30 nucleotides was located close to the putative reaction site in a derivative of a naturally occurring self folding RNA whose crystal structure is known. RNAs whose activity depended on the starting three-dimensional structure were selected with 3'-5' ligation specificity, indicating that the strategy can be used to acquire a variety of catalytic sites and other functional RNA modules. PMID- 15525712 TI - Modified nucleotides at the 5' end of human U2 snRNA are required for spliceosomal E-complex formation. AB - U2 snRNA, a key player in nuclear pre-mRNA splicing, contains a 5'-terminal m3G cap and many internal modifications. The latter were shown in vertebrates to be generally required for U2 function in splicing, but precisely which residues are essential and their role in snRNP and/or spliceosome assembly is presently not clear. Here, we investigated the roles of individual modified nucleotides of HeLa U2 snRNA in pre-mRNA splicing, using a two-step in vitro reconstitution/complementation assay. We show that the three pseudouridines and five 2'O-methyl groups within the first 20 nucleotides of U2 snRNA, but not the m3G cap, are required for efficient pre-mRNA splicing. Individual pseudouridines were not essential, but had cumulative effects on U2 function. In contrast, four of five 2'O-methylations (at positions 1, 2, 12, and 19) were individually required for splicing. The in vitro assembly of 17S U2 snRNPs was not dependent on the presence of modified U2 residues. However, individual internal modifications were required for the formation of the ATP-independent early spliceosomal E complex. Our data strongly suggest that modifications within the first 20 nucleotides of U2 play an important role in facilitating the interaction of U2 with U1 snRNP and/or other factors within the E complex. PMID- 15525713 TI - Serum antibody responses after intradermal vaccination against influenza. AB - BACKGROUND: If found to be safe and immunogenic, reduced doses of influenza vaccine given by the intradermal route could increase the number of available doses of vaccine. METHODS: In an open-label study, we randomly assigned 119 subjects to receive an intradermal injection of trivalent inactivated influenza vaccine, containing 6 mug of hemagglutinin for each antigen (40 percent of the usual dose), and 119 to receive an intramuscular injection of the standard dose of 15 mug of hemagglutinin for each antigen. The two groups were subdivided according to age (18 to 60 years and older than 60 years). RESULTS: Among subjects who were 18 to 60 years of age, serum antibody responses were vigorous and did not differ significantly between the intradermal and intramuscular groups, and all subjects had hemagglutination-inhibition (HAI) titers of at least 1:40. Although the subjects who were older than 60 years of age also had a vigorous antibody response, there was a trend toward a better response in the intramuscular route, but this finding was significant only for antigen to the H3N2 strain. Nevertheless, 100 percent of older subjects in the intramuscular group and 93 percent of such subjects in the intradermal group had an HAI antibody titer to the H3N2 strain of more than 1:40, and 100 percent in each group had a titer of this level for both the H1N1 and B strains. Local pain was significantly more common in the intramuscular group than in the intradermal group among subjects who were 18 to 60 years of age but not among subjects who were over 60 years old. Signs of local inflammation were significantly more common among subjects in the intradermal group than among those in the intramuscular group, in both age groups. CONCLUSIONS: As compared with an intramuscular injection of full-dose influenza vaccine, an intradermal injection of a reduced dose resulted in similarly vigorous antibody responses among persons 18 to 60 years of age but not among those over the age of 60 years. PMID- 15525714 TI - Dose sparing with intradermal injection of influenza vaccine. AB - BACKGROUND: The loss of half the U.S. supply of influenza vaccine due to contamination has created a critical shortage. Dose-sparing strategies that use intradermal delivery of vaccines may be one approach to consider. METHODS: We conducted a randomized, open-label trial outside the influenza season in 100 healthy adults 18 to 40 years of age to compare the immunogenicity and safety of intradermal immunization with influenza vaccine with standard intramuscular immunization. Subjects were randomly assigned to receive either a single intramuscular dose of 0.5 ml of trivalent influenza vaccine, containing at least 15 microg of hemagglutinin per strain, by means of a prefilled syringe or a single intradermal dose of 0.1 ml, containing at least 3 microg of hemagglutinin per strain, by means of a fine-gauge needle; both injections were in the deltoid region. Changes in the hemagglutination-inhibition (HAI) antibody titer were assessed by comparing geometric mean titers and fold increases relative to baseline values and by comparing changes in the seroconversion and seroprotection rates. Local and systemic adverse events were assessed after both types of vaccination. RESULTS: Subjects who received an intradermal injection with one fifth the standard dose of influenza vaccine had increases in the geometric mean HAI titer by a factor of 15.2 for the H1N1 strain in the vaccine, 19.0 for the H3N2 strain, and 12.4 for the B strain on day 21, as compared with respective increases by a factor of 14.9, 7.1, and 15.3 for the intramuscular injection of the standard dose. Seroconversion and seroprotection rates were similar in the two groups on day 21, ranging from 66 to 82 percent and 84 to 100 percent, respectively. Local reactions were significantly more frequent among recipients of intradermal injections than among recipients of intramuscular injections, but such reactions were mild and transient. CONCLUSIONS: In this study of young adults, intradermal administration of one fifth the standard intramuscular dose of an influenza vaccine elicited immunogenicity that was similar to or better than that elicited by intramuscular injection. Intradermal administration could be used to expand the supplies of influenza vaccine, but further studies are needed before this strategy can be recommended for routine use. PMID- 15525715 TI - Intradermal influenza vaccination--can less be more? PMID- 15525716 TI - Images in clinical medicine. Backscatter from lead. PMID- 15525717 TI - Advanced maternal age--how old is too old? PMID- 15525718 TI - Minimal intervention--nurse-midwives in the United States. PMID- 15525719 TI - Nondisjunction--a view from ringside. PMID- 15525720 TI - New genetic insights into Parkinson's disease. PMID- 15525721 TI - Mass treatment with single-dose azithromycin for trachoma. AB - BACKGROUND: Trachoma, caused by repeated ocular infection with Chlamydia trachomatis, is an important cause of blindness. Current recommended dosing intervals for mass azithromycin treatment for trachoma are based on a mathematical model. METHODS: We collected conjunctival swabs for quantitative polymerase-chain-reaction assay of C. trachomatis before and 2, 6, 12, 18, and 24 months after mass treatment with azithromycin in a Tanzanian community in which trachoma was endemic. For ethical reasons, at 6, 12, and 18 months, we gave tetracycline eye ointment to residents who had clinically active trachoma. RESULTS: At baseline, 956 of 978 residents (97.8 percent) received either one oral dose of azithromycin or (if azithromycin was contraindicated) a course of tetracycline eye ointment. The prevalence of infection fell from 9.5 percent before mass treatment to 2.1 percent at 2 months and 0.1 percent at 24 months. The quantitative burden of ocular C. trachomatis infection in the community was 13.9 percent of the pretreatment level at 2 months and 0.8 percent at 24 months. At each time point after baseline, over 90 percent of the total community burden of C. trachomatis infection was found among subjects who had been positive the previous time they were tested. CONCLUSIONS: The prevalence and intensity of infection fell dramatically and remained low for two years after treatment. One round of very-high-coverage mass treatment with azithromycin, perhaps aided by subsequent periodic use of tetracycline eye ointment for persons with active disease, can interrupt the transmission of ocular C. trachomatis infection. PMID- 15525722 TI - Mutations in the glucocerebrosidase gene and Parkinson's disease in Ashkenazi Jews. AB - BACKGROUND: A clinical association has been reported between type 1 Gaucher's disease, which is caused by a glucocerebrosidase deficiency owing to mutations in the glucocerebrosidase gene (GBA), and parkinsonism. We examined whether mutations in the GBA gene are relevant to idiopathic Parkinson's disease. METHODS: A clinic-based case series of 99 Ashkenazi patients with idiopathic Parkinson's disease, 74 Ashkenazi patients with Alzheimer's disease, and 1543 healthy Ashkenazi Jews who underwent testing to identify heterozygosity for certain recessive diseases were screened for the six GBA mutations (N370S, L444P, 84GG, IVS+1, V394L, and R496H) that are most common among Ashkenazi Jews. RESULTS: Thirty-one patients with Parkinson's disease (31.3 percent; 95 percent confidence interval, 22.2 to 40.4 percent) had one or two mutant GBA alleles: 23 were heterozygous for N370S, 4 were heterozygous for 84GG, 3 were homozygous for N370S, and 1 was heterozygous for R496H. Among the 74 patients with Alzheimer's disease, 3 were identified as carriers of Gaucher's disease (4.1 percent; 95 percent confidence interval, 0.0 to 8.5 percent): 2 were heterozygous for N370S, and 1 was heterozygous for 84GG. Ninety-five carriers of Gaucher's disease were identified among the 1543 control subjects (6.2 percent; 95 percent confidence interval, 5.0 to 7.4 percent): 92 were heterozygous for N370S, and 3 were heterozygous for 84GG. Patients with Parkinson's disease had significantly greater odds of being carriers of Gaucher's disease than did patients with Alzheimer's disease (odds ratio, 10.8; 95 percent confidence interval, 3.0 to 46.6; P<0.001) or control subjects (odds ratio, 7.0; 95 percent confidence interval, 4.2 to 11.4; P<0.001). Among the patients with Parkinson's disease, patients who were carriers of Gaucher's disease were younger than those who were not carriers (mean [+/-SD] age at onset, 60.0+/-14.2 years vs. 64.2+/-11.7 years; P=0.04). CONCLUSIONS: Our results suggest that heterozygosity for a GBA mutation may predispose Ashkenazi Jews to Parkinson's disease. PMID- 15525723 TI - Clinical practice. Hypersensitivity to hymenoptera stings. PMID- 15525724 TI - Neonatal brain injury. PMID- 15525725 TI - Images in clinical medicine. Getting the lead out. PMID- 15525726 TI - Clinical problem-solving. Special cure. PMID- 15525727 TI - New steps toward eliminating blinding trachoma. PMID- 15525728 TI - The limits of conscientious objection--may pharmacists refuse to fill prescriptions for emergency contraception? PMID- 15525729 TI - Nevirapine plus zidovudine to prevent mother-to-child transmission of HIV. PMID- 15525730 TI - Recurrent venous thromboembolism in men and women. PMID- 15525731 TI - The antibiotic pipeline. PMID- 15525732 TI - Endothelial cells in B-cell lymphomas. PMID- 15525733 TI - Ciguatera poisoning. PMID- 15525734 TI - Gluten contamination of commercial oat products in the United States. PMID- 15525735 TI - Circumpapillary retinal ridge in the shaken-baby syndrome. PMID- 15525736 TI - A review of complications associated with vertebroplasty and kyphoplasty as reported to the Food and Drug Administration medical device related web site. AB - In 2002, approximately 38,000 vertebroplasties and 16,000 kyphoplasties were performed in the United States. As the use of both modalities for the treatment of vertebral compression fractures has increased, so have questions regarding safety and efficacy. The authors addressed this by reviewing both the current literature and complications data reported to the Food and Drug Administration (FDA) Center for Devices and Radiological Health through the on-line database (http://www.fda.gov/cdrh/maude.html) and through the Office of the Freedom of Information Act at the FDA. Although both procedures are largely safe, the FDA data highlight two main concerns: reactions to the use of acrylic (polymethylmethacrylate) bone cement, including hypotension and, in some cases, death, especially when multiple vertebral levels are treated in one setting; and a possible increased risk with kyphoplasty of pedicle fracture and cord compression. PMID- 15525737 TI - Vertebral augmentation and the limits of interpreting complications reported in the food and drug administration manufacturer and user facility device experience database. PMID- 15525738 TI - Medical treatment of peripheral arterial disease: a comprehensive review. AB - Peripheral arterial disease (PAD) is a common manifestation of atherosclerosis that affects more than 10 million people in the United States. The risk factors associated with PAD are similar to those found in patients with coronary artery disease and cerebrovascular disease. Medical therapy of PAD must include modification of cardiovascular risk factors with application of strict secondary prevention guidelines. For improvement in quality of life, a structured exercise rehabilitation program remains the most effective noninterventional treatment strategy, but it is difficult to employ from economic and patient-compliance perspectives. Newer pharmacologic therapies have demonstrated efficacy in patients with intermittent claudication. Emerging strategies for management of these patients include revascularization and maximal medical therapy for improvement of physical function as well as reduction in risk for subsequent major cardiovascular events. This article will review the clinical data supporting aggressive medical interventions for patients with PAD. PMID- 15525739 TI - Comparison of MELD and Child-Pugh scores to predict survival after chemoembolization for hepatocellular carcinoma. AB - PURPOSE: To compare the value of the Child-Pugh and Model for End-stage Liver Disease (MELD) scores to predict patient survival rates after transarterial chemoembolization (TACE) for hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Eighty-seven patients underwent 169 TACE sessions. Child-Pugh and MELD values were calculated before initial treatment. Survival length was tracked from the date of the first TACE procedure. Transplant recipients were censored from the study at the time of surgery. Child-Pugh and MELD scores as well as bilirubin and albumin levels and International Normalized Ratio were placed in high and low categories defined by their respective medians. Patient survival was compared at 3 months, 6 months, 12 months, and 24 months, and patterns were tested with chi2 or Fisher exact tests. Survival over the entire period was examined with Kaplan Meier analysis and differences were tested with log-rank tests. RESULTS: Mean and median survival times for all patients were 24 and 17 months, respectively. Sixteen patients were censored for transplantation at a mean of 12.9 months. MELD and Child-Pugh scores correlated well with each other (r = 0.68). Child-Pugh score (r = -0.35, P = .04) correlated more strongly with 12-month survival than did MELD score (r = -0.26, P = .12). After high/low score category division, a significantly greater survival difference was predicted by Child-Pugh score (27.2 months vs 10.3 months; P = .03) versus MELD score (27.5 months vs 15.8 months; P = .19). An albumin level greater than 3.4 g/dL was also associated with significantly improved survival (29.3 months vs 10.1 months; P = .0032). Survival differences between high-risk and low-risk groups at the 3-, 6-, 12-, and 24 month intervals were significant for low Child-Pugh scores and for albumin levels greater than 3.4 g/dL. Statistical significance was not approached at any of the time lengths with MELD scores. CONCLUSIONS: Child-Pugh score correlates better than MELD score to overall patient survival and is a better predictor than MELD score of survival at specific time points. Of the components of the Child-Pugh and MELD systems, albumin level is the most useful predictor of survival. PMID- 15525740 TI - Results of a retrospective multicenter trial of the Viatorr expanded polytetrafluoroethylene-covered stent-graft for transjugular intrahepatic portosystemic shunt creation. AB - PURPOSE: To report the results of a multicenter experience with the Viatorr expanded polytetrafluoroethylene-covered stent-graft for transjugular intrahepatic portosystemic shunt (TIPS) creation in which patency and clinical outcome were evaluated. MATERIALS AND METHODS: One hundred consecutive patients with portal hypertension, with a mean age of 52 years (range, 22-86 years), underwent implantation of the Viatorr TIPS stent-graft at one of three hospital centers. The indications for TIPS creation were variceal bleeding (n = 81) and refractory ascites (n = 19). Twenty patients had Child-Pugh class A disease, 46 had class B disease, and 34 had class C disease. Eighty-seven patients underwent de novo TIPS placements, with 13 treated for recurrent TIPS stenosis. Sixty-two patients were available for follow-up portal venography and portosystemic pressure gradient (PSG) measurement commencing 6 months after Viatorr stent-graft placement. RESULTS: The technical success rate was 100%. TIPS creation resulted in an immediate decrease in mean PSG (+/-SD) from 21 mm Hg +/- 6 to 7 mm Hg +/- 3. Acute repeat intervention (within 30 days) was required for portal vein thrombosis (n = 1), continued bleeding (n = 3), and encephalopathy (n = 1). The all-cause 30-day mortality rate was 12%. Two patients developed acute severe refractory encephalopathy, which led to death in one case. New or worsening encephalopathy was identified in 14% of patients. The incidence of recurrent bleeding was 8%. The cumulative survival rate at 1 year was 65%. Sixty-two patients available for venographic follow-up had a mean PSG of 9 mm Hg +/- 5 at a mean interval of 343 days (range, 56-967 days). There were four stent-graft occlusions (6%) and seven hemodynamically significant stenoses (11%), four within the stent-graft and three in the non-stent-implanted hepatic vein. The primary patency rate at 1 year by Kaplan-Meier analysis was 84%. CONCLUSIONS: This retrospective multicenter experience with the Viatorr stent-graft confirms the preliminary findings of other investigators of good technical results and improved patency compared with bare stents. Early mortality and symptomatic recurrence rates are low by historical standards. The theoretical increase in TIPS-related encephalopathy was not demonstrated. Longer-term follow-up will be required to determine whether the additional cost of the Viatorr stent-graft will be offset by reduced surveillance and repeat intervention. PMID- 15525741 TI - Iliac artery stent placement: clinical experience and short-term follow-up regarding a self-expanding nitinol stent. AB - PURPOSE: To evaluate the effectiveness, safety, and short-term patency rates of a self-expandable nitinol stent (JostentSelfX; Abbott Laboratories, Chicago, IL) for treatment of iliac artery stenoses. MATERIALS AND METHODS: In a prospective study, 34 chronic iliac artery stenoses were primarily treated in 27 patients (mean age, 61.7 years +/- 9.4) by placement of a JostentSelfX. Handling and visibility of the stent was scored on a 4-point scale. Before stent placement, within 3 days after stent placement and at 6 months follow-up, the clinical history, clinical stage according to the Rutherford classification, and the ankle brachial index (ABI) were assessed. Intra-arterial digital subtraction angiography including measurement of the mean pressure gradient was performed at the time of stent placement and at 6 months follow-up or in case of recurrence of symptoms. The Wilcoxon signed rank test and Kaplan-Meier method were applied for statistical analysis. RESULTS: The immediate technical success rate was 94%. The clinical success rate immediately after the procedure was 96% and 85% at 6 months follow-up. Stent placement significantly decreased the severity of stenoses as well as the mean pressure gradients and significantly increased the ABI. Handling of the stent was judged good. No misplacement occurred. Foreshortening during deployment was negligible and no stent migration was seen. However, visibility of the stent during fluoroscopy was ranked as moderate to bad. Follow-up at 6 months revealed the cumulative angiographic primary patency rate (<50% restenosis and <10 mm Hg mean pressure gradient) to be 0.96. CONCLUSION: The evaluated self expandable nitinol stent is an effective tool in treating iliac artery stenoses. Apart from poor visibility, handling was good. The angiographic patency rate is comparable with data reported in other stent trials. PMID- 15525742 TI - Cryoplasty for femoropopliteal arterial disease: late angiographic results of initial human experience. AB - PURPOSE: A new form of angioplasty, called cryoplasty, was developed to improve the late results typically associated with percutaneous transluminal angioplasty. Cryoplasty combines the dilation force of percutaneous transluminal angioplasty with the delivery of cold thermal energy to the vessel wall. This study reports the authors' early clinical experience with cryoplasty in patients with femoropopliteal disease. MATERIALS AND METHODS: Fifteen patients with femoropopliteal arterial lesions were treated with cryoplasty (CryoVascular Systems, Los Gatos, CA). Cryoplasty was performed at 6 atm of pressure and delivered at -10 degrees C for 60 seconds. The ankle-brachial index (ABI) was measured at baseline, 24 hours after cryoplasty, and at 1 and 3 months during follow-up. Repeated angiography was performed at 6 and 18 months to determine short-term and late primary patency. RESULTS: Cryoplasty was technically successful in 93% of patients (< 30% residual stenosis and less then grade C dissection). ABIs at baseline were 0.64 +/- 0.08 and improved the day after cryoplasty to 0.95 +/- 0.09 (P < .05). ABIs were well maintained at 1 and 3 months with measurements of 0.94 +/- 0.09 and 0.92 +/- 0.10, respectively (P < .05 vs baseline). Baseline angiographic diameter stenosis improved significantly immediately following cryoplasty (86% +/- 12% to 16% +/- 3%; P < .05). Angiography at 6 months revealed 0% binary restenosis and insignificant change in residual stenosis from the acute cryoplasty results (16 +/- 3% vs 21 +/- 5%; P = NS). Late angiographic follow-up at 14 months +/- 4 demonstrated primary patency of 83.3%. CONCLUSION: Cryoplasty was able to achieve substantial dilation of femoropopliteal lesions with well-preserved late angiographic patency. Cryoplasty represents a potential advance in the field of endovascular medicine. PMID- 15525743 TI - Comparison of sequential versus simultaneous methods of adrenal venous sampling. AB - PURPOSE: To compare two methods of adrenal venous sampling (AVS) in preoperative localization of adrenal lesions in primary hyperaldosteronism. MATERIALS AND METHODS: Twenty-one patients (13 men, eight women) underwent selective adrenal venous sampling between July 2001 and May 2003. One of the 21 patients underwent repeat AVS, for a total of 22 procedures. In half the procedures (n = 11), simultaneous bilateral adrenal venous catheterization and sampling was performed before and after intraprocedural adrenocorticotropic hormone (ACTH) administration; in the remaining half (n = 11), sequential catheterization of the left and right adrenal veins was performed during continuous ACTH infusion 1 hour before and throughout AVS. Chart review provided procedural data, including sampling intervals and aldosterone/cortisol ratios. Patient records provided clinical data, including blood pressure, serum aldosterone levels, and computed tomography and magnetic resonance imaging findings. Surgical pathology reports confirmed unilateral disease but were not applicable to bilateral disease. RESULTS: Selective AVS was completed successfully in 21 of 22 procedures (95%); the unsuccessful sampling was repeated successfully. Disease lateralized in 13 of 22 cases. Simultaneous bilateral AVS localized unilateral disease in seven of eight cases (88%) and was nondiagnostic in one case (13%), with cases confirmed by surgical pathology reports. Sequential bilateral AVS localized unilateral disease in four of four cases (100%) confirmed by surgical pathology reports, with one lost to follow-up. Bilateral disease was diagnosed in six of 22 cases: two of 11 by simultaneous AVS and four of 11 by sequential AVS. Three of 22 cases demonstrated borderline hormone levels that failed to meet the diagnostic threshold for recommended adrenalectomy. Mean elapsed time between acquisition of right and left samples did not differ between simultaneous and sequential AVS (P = .09). Baseline (prestimulation) sampling did not contribute unique diagnostic information in any case and provided contradictory or confounding information in three of 11 simultaneous AVS procedures (27%). CONCLUSIONS: Sequential bilateral catheterization does not compromise the reliability of time-sensitive AVS. Both simultaneous and sequential AVS are adequate studies; however, obtaining baseline prestimulation samples during simultaneous AVS is unnecessary and increases the cost of the procedure. PMID- 15525744 TI - Selective arterial calcium stimulation and hepatic venous sampling in the evaluation of hyperinsulinemic hypoglycemia: potential and limitations. AB - PURPOSE: To describe the potential and limitations of selective arterial calcium stimulation and hepatic venous sampling (ASVS) in the evaluation of patients with hyperinsulinemic hypoglycemia. MATERIALS AND METHODS: Twenty-seven consecutive adult patients with documented hyperinsulinemic hypoglycemia undergoing an ASVS test and histologically confirmed diagnosis were included. RESULTS: By histology, 24 patients were found to have an insulinoma, two had islet cell hyperplasia, and one had insulin-secreting neuroendocrine carcinoma of the liver. ASVS correctly predicted the source of excessive insulin secretion in 26 of 27 (sensitivity, 96%) patients. In two patients with an insulinoma, ASVS had to be repeated for correct diagnosis; in one patient with a glucose-sensitive insulinoma, ASVS was falsely negative. In one patient, a falsely negative ASVS test was caused by diazoxide treatment; in another patient a falsely negative ASVS test occurred because of extraordinarily high baseline insulin levels when the calcium stimulation was performed. CT and/or MR imaging correctly predicted the source of excessive insulin secretion in 59% of patients, the sensitivity of the intraoperative exploration was 88%. CONCLUSIONS: In the authors' experience, the ASVS test is superior to CT/MR imaging and even the intraoperative exploration in identifying the source of excessive insulin secretion in patients with hyperinsulinemic hypoglycemia. To prevent false negative ASVS tests, medications with an influence on insulin secretion have to be discontinued prior to ASVS and two baseline blood samples should be obtained before the calcium stimulation is performed. PMID- 15525745 TI - Extended interval for retrieval of Gunther Tulip filters. AB - PURPOSE: To evaluate the Gunther Tulip vena cava filter with regard to ease of placement, complications, and retrieval over long time periods. MATERIALS AND METHODS: In 53 patients (ratio of men to women, 24:29; mean age, 52.8 years) retrievable Gunther Tulip filters (Vena Cava M Reye Filter Set; William Cook Europe, Denmark) were inserted. Indications included planned major surgery with recent pulmonary embolus or high pulmonary embolus risk (n = 16), extensive ilio femoral thrombus (n = 11), deep vein thrombosis with anticoagulant complications (n = 9), breakthrough pulmonary embolus despite anticoagulant therapy (n = 4), and contraindication to anticoagulant therapy (n = 13). All patients were followed-up for immediate and long-term complications. RESULTS: Fifty-three filters were successfully placed in 52 of 53 patients, yielding a success rate of 98.1%. Nineteen patients underwent attempted retrieval of their filter. Sixteen of 19 retrieval procedures were successful (84%). In three patients, the filter could not be removed on attempted retrieval (extensive filter thrombus in two patients and attachment to the wall in one patient). One patient received two filters, which were both successfully retrieved at a later date. Median implantation time for retrievable filters was 34 days (range, 7-126 days). Mean follow-up for patients with permanent filters was 13 months. Two major complications (pneumothorax and break through pulmonary embolus) and three minor complications (right internal jugular vein thrombosis in two patients and transient Horner's Syndrome in one patient) were recorded. CONCLUSION: Insertion and retrieval of filters is safe and feasible. Preliminary data suggest that Gunther Tulip filter retrieval is feasible over and above the manufacturer's recommended retrieval interval of 14 days. PMID- 15525746 TI - Inferior mesenteric artery embolization before endovascular aneurysm repair: technique and initial results. AB - PURPOSE: To report a single center's technique and initial results in the preoperative embolization of the inferior mesenteric artery (IMA) before endovascular aneurysm repair (EVAR). MATERIALS AND METHODS: Over a 3-year period, 102 patients at a single clinical site, including 86 men and 16 women aged 54-93 years (mean, 75 years), were found to have a patent IMA on computed tomographic (CT) angiography before EVAR. Coil embolization was performed after subselective catheterization with use of microcoils placed in the IMA proximal to the origin of the left colic artery. All patients in whom the IMA was visualized on flush aortography and successfully accessed underwent embolization. One month and 6 months after surgery, results in this cohort were retrospectively compared with those from a similar group of patients who underwent EVAR during the same period. These patients had patent IMAs on preoperative CT angiography but did not undergo embolization as a result of nonvisualization during flush aortography. All patients underwent EVAR with bifurcated modular devices with proximal transrenal fixation. All patients underwent postoperative follow-up with multiphase CT angiography to detect the presence of endoleak. Six-month follow-up data were available for 18 patients who underwent embolization and 54 patients who did not. Change in sac diameter was compared in these patients. RESULTS: Embolization was technically successful in 30 of 32 patients (94%) in whom it was attempted. There were no complications. At 1-month follow-up, five of 30 patients in the embolization group were noted to have a type II endoleak (17%). None of the endoleaks in this group were related to the IMA. The group with patent IMAs who did not undergo preoperative embolization had a 42% incidence of type II endoleak (P < .05). At 6 months after surgery, three of 18 patients who had undergone embolization (17%) had a type II endoleak, compared with 26 of 54 in the other group (48%; P < .05). Among the patients in whom 6-month data were available, mean changes in sac diameter were -5.2 mm (range, -24 to 2 mm) in the embolized group and -2.1 mm (range, -19 to 8 mm) in the nonembolized group. CONCLUSION: These initial results demonstrate that embolization of the IMA with subselective microcoils before EVAR is a safe and effective procedure to reduce the incidence of type II endoleaks. The data also suggest that preoperative embolization of the IMA is associated with greater shrinkage of aneurysm sac diameter at 6 months. PMID- 15525747 TI - Magnetic resonance angiography in the follow-up of distal lower-extremity bypass surgery: comparison with duplex ultrasound and digital subtraction angiography. AB - PURPOSE: The danger of limb loss as a consequence of acute occlusion of infrapopliteal bypasses underscores the requirement for careful patient follow up. The objective of this study was to determine the agreement and accuracy of contrast material-enhanced moving-table magnetic resonance (MR) angiography and duplex ultrasonography (US) in the assessment of failing bypass grafts. In cases of discrepancy, digital subtraction angiography (DSA) served as the reference standard. MATERIALS AND METHODS: MR angiography was performed in 24 consecutive patients with 26 femorotibial or femoropedal bypass grafts. Each revascularized limb was divided into five segments--(i) native arteries proximal to the graft; (ii) proximal anastomosis; (iii) graft course; (iv) distal anastomosis; and (v) native arteries distal to the graft-resulting in 130 vascular segments. Three readers evaluated all MR angiograms for image quality and the presence of failing grafts. The degree of stenosis was compared to the findings of duplex US, and in case of discrepancy, to DSA findings. Two separate analyses were performed with use of DSA only and a combined diagnostic endpoint as the reference standard. RESULTS: Image quality was rated excellent or intermediate in 119 of 130 vascular segments (92%). Venous overlay was encountered in 26 of 130 segments (20%). In only two segments was evaluation of the outflow region not feasible. One hundred seventeen of 130 vascular segments were available for quantitative analysis. In 109 of 117 segments (93%), MR angiography and duplex US showed concordant findings. In the eight discordant segments in seven patients, duplex US overlooked four high-grade stenoses that were correctly identified by MR angiography and confirmed by DSA. Percutaneous transluminal angioplasty was performed in these cases. In no case did MR angiography miss an area of stenosis of sufficient severity to require treatment. Total accuracy for duplex US ranged from 0.90 to 0.97 depending on the reference standard used, whereas MR angiography was completely accurate (1.00) regardless of the standard definition. CONCLUSION: Our data strongly suggest that the accuracy of MR angiography for identifying failing grafts in the infrapopliteal circulation is equal to that of duplex US and superior to that of duplex US in cases of complex revascularization. MR angiography should be included in routine follow-up of patients undergoing infrapopliteal bypass surgery. PMID- 15525748 TI - Liver malignancies: CT-guided interstitial brachytherapy in patients with unfavorable lesions for thermal ablation. AB - PURPOSE: To evaluate computed tomography (CT)-guided brachytherapy in patients with very large liver malignancies or with liver tumors located adjacent to the liver hilum. MATERIALS AND METHODS: In this prospective nonrandomized phase II trial, we treated 20 patients (19 liver metastases and one cholangiocarcinoma) with CT-guided high-dose-rate (HDR) brachytherapy using a (192) Iridium source. All patients demonstrated no functional liver degradation prior to irradiation. Entry criteria were liver tumors > 5 cm (group A, n = 11, no upper limit) or liver tumors < or = 5 cm adjacent to the liver hilum (group B, n = 9). Dose planning for brachytherapy was performed with three dimensional (3D) CT data acquired after percutaneous applicator positioning. Magnetic resonance (MR) imaging follow-up was performed 6 weeks and every 3 months post intervention. Primary endpoints were complications, local tumor control (absence of tumor growth after treatment followed by shrinkage of the lesion starting at 6 months) and progression-free survival. RESULTS: The mean tumor diameter was 7.7 cm (range, 5.5-10.8 cm) in group A, 3.6 cm (range, 2.2-4.9 cm) in group B. On average, a minimal dose of 17 Gy in the target volume was applied (range, 12-25 Gy). Severe side effects were recorded in two patients (10%). One patient demonstrated an obstructive jaundice caused by tumor edema after irradiation of a metastasis adjacent to the bile duct bifurcation. One patient developed intra abdominal hemorrhage that was treated by a single blood transfusion and has ceased. We frequently encountered moderate increases of liver enzymes (70% of patients) and bilirubin (50% of patients) without clinical symptoms and thus considered to be insignificant. The median follow-up was 13 months. In group A (tumors > 5 cm), primary local tumor control after 6 and 12 months was 74% and 40%, respectively; in group B, it was 100% and 71%, respectively. All but one local recurrence (in a patient with diffuse tumor progression) were successfully treated during another CT-guided brachytherapy leading to a primary assisted local control of 93% after 12 months. CONCLUSION: CT-guided brachytherapy based on individual dose plans and 3D CT data sets generated encouraging results in large liver malignancies as well as in tumors located adjacent to the liver hilum. PMID- 15525749 TI - Efficient inhibition of in-stent restenosis by controlled stent-based inhibition of elastase: a pilot study. AB - PURPOSE: It is proposed that local elastase inhibition could suppress the extracellular matrix (ECM) degradation and subsequent smooth muscle cell migration and limit subsequent in-stent restenosis. This study evaluated the effect of stent-based controlled elastase inhibition on restenosis after stent implantation in a rabbit model. MATERIALS AND METHODS: Biodegradable microspheres containing the potent elastase inhibitor alpha-1-antitrypsin (AAT) were prepared. Daily release of AAT from the microspheres was confirmed in vitro. The microspheres were loaded into stents with an abluminal polymer reservoir. Implantation of the stent with AAT microspheres and blank microspheres (control) was performed in the abdominal aortae of six rabbits in each group. After stent deployment, all stents were overdilated to 125% diameter. Stent-implanted arteries were harvested after 7 days (n = 3 each) or 28 days (n = 3 each). To assess the effect of local delivery of AAT, elastase activity and elastin content of the stent-implanted aortae were analyzed. As an endpoint, intima-to-media (I/M) ratio was determined in the 7-day and 28-day specimens. RESULTS: Significant inhibition of elastase was confirmed in treated vessels versus controls at 7 days after stent implantation (P < .05). This reduction in elastase activity was sufficient to afford early and late reduction of in-stent neointima. Plaque progression in the 28-day specimens decreased to 67% with elastase inhibition relative to controls (P < .05). CONCLUSION: Stent-based controlled release of elastase inhibitor may significantly reduce ECM degradation and might limit in-stent restenosis. PMID- 15525750 TI - An in vitro analysis of a carotid artery stent with a protective porous membrane. AB - PURPOSE: To prove the effectiveness of a new stent concept with integrated protection (MembraX [MX]) by comparing it with five cerebral protection devices designed for carotid angioplasty in an in vitro model. MATERIALS AND METHODS: Two simulation series of embolization from carotid angioplasty have been performed. In the first series, polyvinyl-alcohol particles (150-250 microm [small], 355-500 microm [medium], 710-1000 microm [large]; 5 mg each) were injected into a silicone flow model simulating the aortic arch with a carotid bifurcation. The particles were injected proximally to the partially deployed MX stent or one of the following protection devices: Angioguard (AG), FilterWire EX (EX), Trap, Neuroshield (NS), or GuardWire Plus (GW). Particles evading the protection device were caught in a filter at the end of the flow model and weighed. In the second series, human plaque material (8-12 particles; total weight 6.09 +/- 0.01 mg; 500 1500 microm) was injected into the model with the respective devices. MX was compared with the AG, EX, Trap, and NS devices. RESULTS: MX had the most effective overall filtration performance for polyvinyl alcohol particles in the effluent of the internal carotid artery (ICA; 0.43 mg, 2.9%), compared with NS (0.53 mg, 3.5%), GW (1.10 mg, 7.0%), EX and AG (1.18 and 1.21 mg, respectively; 7.8% and 8.0%), and Trap (1.24 mg, 8.2%). MX performed best for the small particles (2.0% passed particles into ICA; P < .05 compared with all). Human plaque material was retained best in the in vitro model by MX (0.0%), followed by NS (0.8%), EX (1.3%), Trap (2.6%), and AG (4.4%). CONCLUSIONS: In vitro, none of the tested devices had the ability to prevent embolization completely. Comparing current designs, the MX device captured the highest percentage of the three different particle groups. Tested with human plaque emboli, MX performed effectively in filtering the particles in the ICA. PMID- 15525751 TI - Reversible neuropathy caused by overuse following radiofrequency ablation of metastatic pelvic lesions. AB - Adverse events following radiofrequency ablation of skeletal metastases are uncommon. This report describes two patients who developed a delayed neuropathy following radiofrequency ablation of pelvic metastases. Both patients had significant pain relief and normal neurological examinations following the procedure. Each patient's neuropathy developed following acute significant increases of activity or stress on surrounding tissues allowed by the pain relief from therapy. Both patients completely recovered after a course of corticosteroids. PMID- 15525752 TI - Inhaled nitric oxide as an adjunct to suction thrombectomy for pulmonary embolism. AB - Pulmonary suction thrombectomy can be a successful interventional tool in the treatment of pulmonary thromboembolism. Removal of clot burden typically results in prompt recovery of hemodynamic stability and improved oxygenation. However, in rare cases, clot removal does not sufficiently improve the clinical situation. Herein, two patients with massive pulmonary thromboembolism are presented whose condition improved only after they received nitric oxide as an adjunct to pulmonary suction thrombectomy. The treatment with this inhalable vasodilator was based on the hypothesis that prolonged ischemia had induced microcirculatory vasospasm, persistent after removal of the central clot. PMID- 15525753 TI - Salvage of endoluminal exclusion of an anastomotic arch aneurysm with a "kissing" carotid stent. AB - Traditional open repair of thoracic aortic aneurysms remains a high-risk surgical procedure. Thoracic aneurysm stent-graft repair is evolving but has promise as a minimally invasive treatment option. Herein a case is presented of an anastomotic arch aneurysm treated with a custom-made thoracic stent-graft after a left subclavian artery-to-carotid artery transposition. Proximal extension for a type I endoleak necessitated a bailout procedure with a "kissing" left common carotid artery bare stent. PMID- 15525754 TI - Transthoracic percutaneous radiofrequency ablation for liver tumors in the hepatic dome. AB - Computed tomography (CT)-guided transthoracic radiofrequency ablation was performed for nine liver tumors of eight patients, which were located in the hepatic dome and undetectable by ultrasound (US). A total 11 sessions of ablation were performed. Complications were noted in five sessions (45%) and no complications were noted in six sessions (55%). Pneumothorax was noted in five sessions (45%), including two sessions (18%) with major pneumothorax requiring a chest tube. Major complications were seen in two sessions (18%), major pneumothorax and both major pneumothorax and moderate pleural effusion, respectively. CT-guided transthoracic radiofrequency ablation may be an alternative for treatments of liver tumor in the hepatic dome. PMID- 15525755 TI - Percutaneous ultrasound-guided biopsy in the definitive diagnosis of osteosarcoma. AB - Thirty-three patients with 35 bone lesions suspicious for osteosarcoma underwent image-guided biopsy. Of those 35 biopsies, 12 were performed with use of fluoroscopy or computed tomography (CT) guidance to target the osseous abnormality. Diagnostic magnetic resonance imaging revealed a soft tissue component in 23 cases; in these cases, biopsies were performed with use of ultrasonography (US) to target the soft tissue component of the tumor. Of the 35 tumors in which biopsy was performed, 33 (94%) were definitively diagnosed as osteosarcoma by image-guided biopsy. All 23 US-guided biopsies resulted in definitive diagnosis. Two of the 12 fluoroscopy- or CT-guided biopsies (17%) were inconclusive. PMID- 15525756 TI - Percutaneous thrombin injection treatment of a profunda femoris pseudoaneurysm after femoral neck fracture. PMID- 15525757 TI - Identification of amino acid residues in the insect sodium channel critical for pyrethroid binding. AB - The voltage-gated sodium channel is the primary target site of pyrethroids, which constitute a major class of insecticides used worldwide. Pyrethroids prolong the opening of sodium channels by inhibiting deactivation and inactivation. Despite numerous attempts to characterize pyrethroid binding to sodium channels in the past several decades, the molecular determinants of the pyrethroid binding site on the sodium channel remain elusive. Here, we show that an F-to-I substitution at 1519 (F1519I) in segment 6 of domain III (IIIS6) abolished the sensitivity of the cockroach sodium channel expressed in Xenopus laevis oocytes to all eight structurally diverse pyrethroids examined, including permethrin and deltamethrin. In contrast, substitution by tyrosine or tryptophan reduced the channel sensitivity to deltamethrin only by 3- to 10-fold, indicating that an aromatic residue at this position is critical for the interaction of pyrethroids with sodium channels. The F1519I mutation, however, did not alter the action of two other classes of sodium channel toxins, batrachotoxin (a site 2 toxin) and Lqhalpha-IT (a site 3 toxin). Schild analysis using competitive interaction of pyrethroid-stereospecific isomers demonstrated that the F1519W mutation and a previously known pyrethroid-resistance mutation, L993F in IIS6, reduced the binding affinity of 1S-cis-permethrin, an inactive isomer that shares the same binding site with the active isomer 1R-cis-permethrin. Our results provide the first direct proof that Leu993 and Phe1519 are part of the pyrethroid receptor site on an insect sodium channel. PMID- 15525758 TI - Kv2.1 channel activation and inactivation is influenced by physical interactions of both syntaxin 1A and the syntaxin 1A/soluble N-ethylmaleimide-sensitive factor 25 (t-SNARE) complex with the C terminus of the channel. AB - Kv2.1, the prevalent delayed-rectifier K(+) channel in neuroendocrine and endocrine cells, was suggested previously by our group to be modulated in islet beta-cells by syntaxin 1A (Syx) and soluble N-ethylmaleimide-sensitive factor attachment protein-25 (SNAP-25). We also demonstrated physical interactions in neuroendocrine cells between Kv2.1, Syx, and SNAP-25, characterized their effects on Kv2.1 activation and inactivation in Xenopus laevis oocytes, and suggested that they pertain to the assembly/disassembly of the Syx/SNAP-25 (t-SNARE) complex. In the present work, we established the existence of a causal relationship between the physical and the functional interactions of Syx with the Kv2.1 channel using three different peptides that compete with the channel for binding of Syx when injected into oocytes already coexpressing Syx with Kv2.1 in the plasma membrane: one peptide corresponding to the Syx-binding region on the N type Ca(2+) channel, and two peptides corresponding to Syx-binding regions on the Kv2.1 C terminus. All peptides reversed the effects of Syx on Kv2.1, suggesting that the hyperpolarizing shifts of the steady-state inactivation and activation of Kv2.1 caused by Syx result from cell-surface protein-protein interactions and point to participation of the C terminus in such an interaction. In line with these findings, the effects of Syx were dissipated by partial deletions of the C terminus. Furthermore, the t-SNARE complex was shown to bind to the Kv2.1 C terminus, and its effects on the inactivation of Kv2.1 were dissipated by partial deletions of the C terminus. Taken together, these findings suggest that physical interactions of both Syx and the t-SNARE complex with the C terminus of Kv2.1 are involved in channel regulation. PMID- 15525759 TI - The S6KII (rsk) gene of Drosophila melanogaster differentially affects an operant and a classical learning task. AB - In an attempt to dissect classical and operant conditioning in Drosophila melanogaster, we have isolated the gene for ribosomal S6 kinase II (S6KII). This enzyme is part of a family of serine-threonine kinases that in mammals have been implicated in the MAPK (mitogen-activated protein kinase) signaling cascade controlling (among other processes) synaptic plasticity (long-term potentiation/long-term depression) and memory formation. The human homolog rsk2 has been linked to mental retardation (Coffin-Lowry syndrome). Mutant analysis in Drosophila shows that S6KII serves different functions in operant place learning and classical (pavlovian) olfactory conditioning. Whereas in the null mutant only pavlovian olfactory learning is affected, a P-element insertion mutant reducing the amount of S6KII only affects operant place learning. A mutant lacking part of the N-terminal kinase domain and performing poorly in both learning tasks is dominant in the operant paradigm and recessive in the pavlovian paradigm. The behavioral defects in the pavlovian task can be rescued by the genomic S6KII transgene. Overexpression of S6KII in wild type has a dominant-negative effect on the operant task that is rescued by the null mutant, whereas in the pavlovian task overexpression may even enhance learning performance. PMID- 15525760 TI - Visualizing synaptic ribbons in the living cell. AB - Visual and auditory information is encoded by sensory neurons that tonically release neurotransmitter at high rates. The synaptic ribbon is an essential organelle in nerve terminals of these neurons. Its precise function is unknown, but if the ribbon could be visualized in a living terminal, both its own dynamics and its relation to calcium and vesicle dynamics could be studied. We designed a short fluorescent peptide with affinity for a known binding domain of RIBEYE, a protein unique to the ribbon. When introduced via a whole-cell patch pipette, the peptide labeled structures at the presynaptic plasma membrane of ribbon-type terminals. The fluorescent spots match in size, location, number, and distribution the known features of synaptic ribbons. Furthermore, fluorescent spots mapped by confocal microscopy directly match the ribbons identified by electron microscopy in the same cell. Clearly the peptide binds to the synaptic ribbon, but even at saturating concentrations it affects neither the morphology of the ribbon nor its tethering of synaptic vesicles. It also does not inhibit exocytosis. Using the peptide label, we observed that the ribbon is immobile over minutes and that calcium influx is concentrated at the ribbon. Finally, we find that each ribbon in a retinal bipolar cell contains approximately 4000 molecules of RIBEYE, indicating that it is the major component of the synaptic ribbon. PMID- 15525761 TI - Brain-derived neurotrophic factor induces mammalian target of rapamycin-dependent local activation of translation machinery and protein synthesis in neuronal dendrites. AB - In neurons, perisynaptic or dendritic translation is implicated in synapse-wide alterations of function and morphology triggered by neural activity. The molecular mechanisms controlling local translation activation, however, have yet to be elucidated. Here, we show that local protein synthesis and translational activation in neuronal dendrites are upregulated by brain-derived neurotrophic factor (BDNF) in a rapamycin and small interfering RNA specific for mammalian target of rapamycin (mTOR)-sensitive manner. In parallel, BDNF induced the phosphorylation of tuberin and the activation of mTOR in dendrites and the synaptoneurosome fraction. mTOR activation stimulated translation initiation processes involving both eIF4E/4E-binding protein (4EBP) and p70S6 kinase/ribosomal S6 protein. BDNF induced phosphorylation of 4EBP in isolated dendrites. Moreover, local puff application of BDNF to dendrites triggered S6 phosphorylation in a restricted area. Taken together, these data indicate that mTOR-dependent translation activation is essential for the upregulation of local protein synthesis in neuronal dendrites. PMID- 15525762 TI - Electrical coupling among irregular-spiking GABAergic interneurons expressing cannabinoid receptors. AB - Anatomical studies have shown that the G-protein-coupled cannabinoid receptor-1 (CB1) is selectively expressed in a subset of GABAergic interneurons. It has been proposed that these cells regulate rhythmic activity and play a key role mediating the cognitive actions of marijuana and endogenous cannabinoids. However, the physiology, anatomy, and synaptic connectivity of neocortical CB1 expressing interneurons remain poorly studied. We identified a population of CB1 expressing interneurons in layer II/III in mouse neocortical slices. These cells were multipolar or bitufted, had a widely extending axon, and exhibited a characteristic pattern of irregular spiking (IS) in response to current injection. CB1-expressing-IS (CB1-IS) cells were inhibitory, establishing GABAA receptor-mediated synapses onto pyramidal cells and other CB1-IS cells. Recently, electrical coupling among other classes of cortical interneurons has been shown to contribute to the generation of rhythmic synchronous activity in the neocortex. We therefore tested whether CB1-IS interneurons are interconnected via electrical synapses using paired recordings. We found that 90% (19 of 21 pairs) of simultaneously recorded pairs of CB1-IS cells were electrically coupled. The average coupling coefficient was 6%. Signaling through electrical synapses promoted coordinated firing among CB1-IS cells. Together, our results identify a population of electrically coupled CB1-IS GABAergic interneurons in the neocortex that share a unique morphology and a characteristic pattern of irregular spiking in response to current injection. The synaptic interactions of these cells may play an important role mediating the cognitive actions of cannabinoids and regulating coherent neocortical activity. PMID- 15525763 TI - Cytokine-induced activation of signal transducer and activator of transcription in photoreceptor precursors regulates rod differentiation in the developing mouse retina. AB - Ciliary neurotrophic factor (CNTF) exhibits multiple biological effects during vertebrate retinogenesis, including regulation of photoreceptor cell differentiation. In the early postnatal mouse retina, CNTF induces rapid and transient phosphorylation of signal transducer and activator of transcription (STAT) 1 and STAT3 and the extracellular signal-regulated kinase (ERK). Although both proliferating progenitor cells and postmitotic neurons respond directly to cytokine signals, CNTF elicits distinct phosphorylation patterns of STAT3 and ERK. CNTF stimulation induces low levels of STAT3 phosphorylation in progenitors and differentiated neurons but a robust STAT3 activation among postmitotic photoreceptor precursors expressing the cone-rod homeobox gene Crx and newly differentiated rod photoreceptors. In contrast, CNTF causes preferential phosphorylation of ERK in progenitor cells and photoreceptor precursors. Inhibition of the cytokine receptor gp130 using neutralizing antibodies reveals that gp130 is required for both CNTF-induced STAT3 and ERK phosphorylation. Perturbation of STAT signaling by a STAT inhibitor peptide or a dominant-negative STAT3 mutant causes enhanced production of rod photoreceptors in the absence of exogenous cytokines, whereas inhibiting ERK activation by a MEK (mitogen activated protein kinase kinase)-specific inhibitor has no effect on rod photoreceptor differentiation in vitro. Furthermore, disrupting the function of epidermal growth factor (EGF) receptors, which modulate rod development in vivo, indicates that the EGF family of ligands does not mediate the inhibitory effect of cytokine on rod differentiation. These results demonstrate that cytokine signal transduction is dynamic and heterogeneous in the developing retina, and that endogenous ligand-induced STAT activation in retinal progenitor and/or photoreceptor precursor cells plays an important role in regulating photoreceptor development. PMID- 15525764 TI - An early broad competence of motoneurons to express ER81 is later sculpted by the periphery. AB - The ETS transcription factor ER81 is expressed in sensory neurons and motoneurons that innervate the adductor and femorotibialis muscles in chick hindlimb and is essential for the development of monosynaptic connections between these two populations of neurons. Neurons need a signal(s) from limb bud mesoderm to initiate ER81 expression. It is not known whether the mature expression pattern arises because adductor and femorotibialis motoneurons are uniquely competent to respond to peripheral signals and express ER81, or whether all motoneurons are competent to express ER81, but normally only adductor and femorotibialis motoneurons are exposed to the requisite activating signal. To investigate these possibilities, we examined ER81 expression in motoneurons that encountered limb tissue surgically mismatched with their target identity at stages after motor pool identities are established. We found that ER81 expression was not invariably linked to motor pool identity or target innervation and was more malleable in later-born femorotibialis motoneurons than in earlier-born adductor motoneurons. We also found that ER81 expression is regulated differently in sensory neurons and motoneurons. Most striking was the observation that motoneurons caudal to the normal adductor and femorotibialis pools could express ER81 when exposed to the appropriate peripheral signals, although this competence did not extend through the entire lumbosacral (LS) region. Thus, it appears that a prepattern of competence to express ER81 is established in early LS motoneurons, most likely in concert with their target identity, and that the expression domains of motoneurons are subsequently refined by peripheral signals at later stages. PMID- 15525765 TI - Superparamagnetic iron oxide-labeled Schwann cells and olfactory ensheathing cells can be traced in vivo by magnetic resonance imaging and retain functional properties after transplantation into the CNS. AB - Schwann cell (SC) and olfactory ensheathing cell (OEC) transplantation has been shown experimentally to promote CNS axonal regeneration and remyelination. To advance this technique into a clinical setting it is important to be able to follow the fates of transplanted cells by noninvasive imaging. Previous studies, using complex modification processes to enable uptake of contrast agents, have shown that cells labeled in vitro with paramagnetic contrast agents transplanted into rodent CNS can be visualized using magnetic resonance imaging (MRI). Here we show that SCs and OECs efficiently internalize dextran-coated superparamagnetic iron oxide (SPIO) from the culture medium by fluid phase pinocytosis. After transplantation into focal areas of demyelination in adult rat spinal cord both transplanted SPIO-labeled SCs and OECs produce a signal reduction using T(2) weighted MRI in anesthetized rats that persists for up to 4 weeks. Although signal reduction was discernable after transplantation of unlabelled cells, this is nevertheless distinguishable from that produced by transplanted labeled cells. The region of signal reduction in SPIO-labeled cell recipients correlates closely with areas of remyelination. Because the retention of functional integrity by labeled cells is paramount, we also show that SPIO-labeled SCs and OECs are able to myelinate normally after transplantation into focal areas of demyelination. These studies demonstrate the feasibility of noninvasive imaging of transplanted SCs and OECs and represent a significant step toward the clinical application of promising experimental approaches. PMID- 15525766 TI - Entorhinal cortex lesions disrupt the relational organization of memory in monkeys. AB - Recent accounts suggest that the hippocampal system critically supports two central characteristics of episodic memory: the ability to establish and maintain representations for the salient relationships between experienced events (relational representation) and the capacity to flexibly manipulate memory (flexible memory expression). To test this proposal in monkeys, intact controls and subjects with bilateral aspiration lesions of the entorhinal cortex were trained postoperatively on two standard memory tasks, delayed nonmatchingto sample (DNMS) and two-choice object discrimination (OD) learning, and three procedures intended to emphasize relational representation and flexible memory expression: a paired associate (PA) task, a transitive inference (TI) test of learning and memory for hierarchical stimulus relationships, and a spatial delayed recognition span (SDRS) procedure. The latter assessments each included critical "probe" tests that asked monkeys to evaluate the relationships among previously learned stimuli presented in novel combinations. Subjects with entorhinal cortex lesions scored as accurately as controls on all phases of DNMS and OD, procedures that can be solved on the basis of memory for individual stimuli. In contrast, experimental monkeys displayed deficits relative to controls on all phases of the PA, TI, and SDRS tasks that emphasized the flexible manipulation of memory for the relationships between familiar items. Together, the findings support the conclusion that the primate hippocampal system critically enables the relational organization of declarative memory. PMID- 15525767 TI - Activity affects dendritic shape and synapse elimination during steroid controlled dendritic retraction in Manduca sexta. AB - Insect metamorphosis is a compelling example for dendritic and synaptic remodeling as larval and adult behaviors place distinct demands on the CNS. During the metamorphosis of the moth, Manduca sexta, many larval motoneurons are remodeled to serve a new function in the adult. During late larval life, steroid hormones trigger axonal and dendritic regression as well as larval synapse elimination. These regressive events are accompanied by stereotypical changes in motor behavior during the so-called wandering stages. Both normally occurring changes in dendritic shape and in motor output have previously been analyzed quantitatively for the individually identified motoneuron MN5. This study tested whether activity affected steroid-induced dendritic regression and synapse disassembly in MN5 by means of chronically implanted extracellular electrodes. Stimulating MN5 in vivo in intact, normally developing animals during a developmental period when it usually shows no activity significantly slowed the regression of high-order dendrites. Both physiological and anatomical analysis demonstrated that reduced dendritic regression was accompanied by a significant reduction in larval synapse disassembly. Therefore, steroid-induced alterations of dendritic shape and synaptic connectivity are modified by activity-dependent mechanisms. This interaction might be a common mechanism for rapid adjustments of rigid, inflexible, hormonal programs. PMID- 15525768 TI - Microglial phagocytosis of fibrillar beta-amyloid through a beta1 integrin dependent mechanism. AB - Microglia are the principle immune effector and phagocytic cells in the CNS. These cells are associated with fibrillar beta-amyloid (fAbeta)-containing plaques found in the brains of Alzheimer's disease (AD) patients. The plaque associated microglia undergo a phenotypic conversion into an activated phenotype and are responsible for the development of a focal inflammatory response that exacerbates and accelerates the disease process. Paradoxically, despite the presence of abundant activated microglia in the brain of AD patients, these cells fail to mount a phagocytic response to Abeta deposits but can efficiently phagocytose Abeta fibrils and plaques in vitro. We report that exposure of microglia to fAbeta in vitro induces phagocytosis through mechanisms distinct from those used by the classical phagocytic receptors, the Ig receptors (FcRgammaI and FcgammaRIII) or complement receptors. Microglia interact with fAbeta through a recently characterized Abeta cell surface receptor complex comprising the B-class scavenger receptor CD36, alpha6beta1 integrin, and CD47 (integrin-associated protein). Antagonists specific for each component of the receptor complex blocks fAbeta-stimulated phagocytosis. These data demonstrated that engagement of this ensemble of receptors is required for induction of phagocytosis. The phagocytic response stimulated by this receptor complex is driven principally by a beta(1) integrin-linked process that is morphologically and mechanistically distinct from the classical type I and type II phagocytic mechanisms. These data provide evidence for phagocytic uptake of fAbeta through a receptor-mediated, nonclassical phagocytic mechanism. PMID- 15525769 TI - The kinetic profile of intracellular calcium predicts long-term potentiation and long-term depression. AB - Efficiency of synaptic transmission within the neocortex is regulated throughout life by experience and activity. Periods of correlated or uncorrelated presynaptic and postsynaptic activity lead to enduring changes in synaptic efficiency [long-term potentiation (LTP) and long-term depression (LTD), respectively]. The initial plasticity triggering event is thought to be a precipitous rise in postsynaptic intracellular calcium, with higher levels inducing LTP and more moderate levels inducing LTD. We used a pairing protocol in visual cortical brain slices from young guinea pigs with whole-cell recording and calcium imaging to compare the kinetic profiles of calcium signals generated in response to individual pairings along with the cumulative calcium wave and plasticity outcome. The identical pairing protocol applied to layer 2/3 pyramidal neurons results in different plasticity outcomes between cells. These differences are not attributable to variations in the conditioning protocol, cellular properties, inter-animal variability, animal age, differences in spike timing between the synaptic response and spikes, washout of plasticity factors, recruitment of inhibition, or activation of different afferents. The different plasticity outcomes are reliably predicted by individual intracellular calcium transients in the dendrites after the first few pairings. In addition to the differences in the individual calcium transients, the cumulative calcium wave that spreads to the soma also has a different profile for cells that undergo LTP versus LTD. We conclude that there are biological differences between like-type cells in the dendritic calcium signals generated by coincident synaptic input and spiking that determine the sign of the plasticity response after brief associations. PMID- 15525770 TI - Deletion of N-type calcium channels alters ethanol reward and reduces ethanol consumption in mice. AB - N-type calcium channels are modulated by acute and chronic ethanol exposure in vitro at concentrations known to affect humans, but it is not known whether N type channels are important for behavioral responses to ethanol in vivo. Here, we show that in mice lacking functional N-type calcium channels, voluntary ethanol consumption is reduced and place preference is developed only at a low dose of ethanol. The hypnotic effects of ethanol are also substantially diminished, whereas ethanol-induced ataxia is mildly increased. These results demonstrate that N-type calcium channels modulate acute responses to ethanol and are important mediators of ethanol reward and preference. PMID- 15525771 TI - Modulation of an afterhyperpolarization by the substantia nigra induces pauses in the tonic firing of striatal cholinergic interneurons. AB - Striatal cholinergic interneurons, also known as tonically active neurons (TANs), acquire a pause in firing during learning of stimulus-reward associations. This pause response to a sensory stimulus emerges after repeated pairing with a reward. The conditioned pause is dependent on dopamine from the substantia nigra, but its underlying cellular mechanism is unknown. Using in vivo intracellular recording, we found that both subthreshold and suprathreshold depolarizations in cholinergic interneurons induced a prolonged after-hyperpolarization (AHP) associated with a pause in their tonic firing. The AHP duration was dependent on the level of depolarization, whether elicited by intracellular current injection or by activation of excitatory inputs from the cortex. High-frequency stimulation of the substantia nigra induced potentiation of the cortically evoked excitation and increased the prolonged AHP after the stimulus. These findings from anesthetized animals suggest that a substantia nigra-induced AHP produces stimulus-associated firing pauses in cholinergic interneurons. This mechanism may underlie the acquisition of the pause response in TANs recorded from behaving animals during learning. PMID- 15525772 TI - Developmental roles of p73 in Cajal-Retzius cells and cortical patterning. AB - To examine the role of the p53 homolog p73 in brain development, we studied p73-/ , p73+/-, E2F1-/-, and reeler mutant mice. p73 in developing brain is expressed in Cajal-Retzius (CR) cells, the cortical hem, and the choroid plexus. p73 expressing CR cells are lost in p73-/- embryos, although Reelin is faintly expressed in the marginal zone. Ectopic neurons in the p73-/- preplate and cortical hem at embryonic day 12 implicate p73 in the early developmental program of the cortex; however, preplate partition and early cortical plate formation are not disturbed. Postnatal p73-/- mice show a mild hypoplasia of the rostral cortex and a severely disrupted architecture of the posterior telencephalon. In the developing p73-/- hippocampus, the most striking abnormality is the absence of the hippocampal fissure, suggesting a role of p73 in cortical folding. p73+/- mice have a less severe cortical phenotype; they display a dorsal shift of the entorhinal cortex and a reduced size of occipital and posterior temporal areas, which acquire entorhinal-like features such as Reelin-positive cells in layer II. CR cells appear unaffected by heterozygosity. We relate the malformations of the posterior pole in p73 mutant mice to alterations of p73 expression in the cortical hem and suggest that p73 forms part of an early signaling network that controls neocortical and archicortical regionalization. In mice deficient for the transcription factor E2F1, a main activator of the TAp73 (transactivating p73) isoform, we find a defect of the caudal cortical architecture resembling the p73+/- phenotype along with reduced TAp73 protein levels and propose that an E2F1 TAp73 dependent pathway is involved in cortical patterning. PMID- 15525773 TI - Glial nitric oxide-mediated long-term presynaptic facilitation revealed by optical imaging in rat spinal dorsal horn. AB - We investigated a presynaptic form of long-term potentiation (LTP) in horizontal slices of the rat spinal cord by visualizing presynaptic and postsynaptic excitation with a voltage-sensitive dye. To record presynaptic excitation, we stained primary afferent fibers anterogradely from the dorsal root. A single pulse test stimulation of C fiber-activating strength to the dorsal root elicited action potential (AP)-like or compound AP-like optical signals throughout the superficial dorsal horn. After conditioning (240 pulses at 2 Hz for 2 min), the presynaptic excitation was augmented. Furthermore, new excitation was elicited in the areas that were silent before conditioning. For postsynaptic recording, projection neurons in spinal lamina I were stained retrogradely from the periaqueductal gray in the brain stem. The test stimulation elicited AP-like or EPSP-like optical signals in the stained neurons. After conditioning, the EPSP like responses were augmented, and previously silent neurons were converted to active ones. Results obtained with a nitric oxide (NO) donor, NO synthase inhibitors, metabotropic glutamate receptor (mGluR) agonist and mGluR1 antagonist, and a glial metabolism inhibitor suggest that after conditioning, presynaptic excitation is facilitated by NO released from glial cells via the activation of mGluR1. The results also indicate the possible presence of additional presynaptic and postsynaptic mechanism(s) for the LTP induction. Activity-dependent LTP of nociceptive afferent synaptic transmission in the spinal cord is believed to underlie central sensitization after inflammation or nerve injury. This glial NO-mediated control of presynaptic excitation may contribute to the induction at least in part. PMID- 15525774 TI - Spiral waves in disinhibited mammalian neocortex. AB - Spiral waves are a basic feature of excitable systems. Although such waves have been observed in a variety of biological systems, they have not been observed in the mammalian cortex during neuronal activity. Here, we report stable rotating spiral waves in rat neocortical slices visualized by voltage-sensitive dye imaging. Tissue from the occipital cortex (visual) was sectioned parallel to cortical lamina to preserve horizontal connections in layers III-V (500-mum thick, approximately 4 x 6 mm(2)). In such tangential slices, excitation waves propagated in two dimensions during cholinergic oscillations. Spiral waves occurred spontaneously and alternated with plane, ring, and irregular waves. The rotation rate of the spirals was approximately 10 turns per second, and the rotation was linked to the oscillations in a one-cycle- one-rotation manner. A small (<128 mum) phase singularity occurred at the center of the spirals, about which were observed oscillations of widely distributed phases. The phase singularity drifted slowly across the tissue ( approximately 1 mm/10 turns). We introduced a computational model of a cortical layer that predicted and replicated many of the features of our experimental findings. We speculate that rotating spiral waves may provide a spatial framework to organize cortical oscillations. PMID- 15525776 TI - Phasic activation of monkey locus ceruleus neurons by simple decisions in a forced-choice task. AB - The noradrenergic locus ceruleus (LC) system has been implicated in several behavioral functions, most notably, response to salient sensory events. Here, we provide new evidence indicating a role in the execution of responses associated with simple decisions. We examined impulse activity of monkey LC neurons during performance of a forced-choice discrimination task. The timing of LC activity more closely tracked behavioral responses than stimulus presentation. LC neurons were phasically activated preceding behavioral responses for both correct and incorrect identifications but were not activated by stimuli that failed to elicit lever responses nor by nontask-related lever movements. We hypothesize that the LC responds to the outcome of task-related decision processes, facilitating their influence on overt behavior. This role of the LC in regulating the behavioral outcome of decisional processes contrasts with more traditional views of LC responses as primarily related to sensory processes. PMID- 15525775 TI - Late calcium EDTA rescues hippocampal CA1 neurons from global ischemia-induced death. AB - Transient global ischemia induces a delayed rise in intracellular Zn2+, which may be mediated via glutamate receptor 2 (GluR2)-lacking AMPA receptors (AMPARs), and selective, delayed death of hippocampal CA1 neurons. The molecular mechanisms underlying Zn2+ toxicity in vivo are not well delineated. Here we show the striking finding that intraventricular injection of the high-affinity Zn2+ chelator calcium EDTA (CaEDTA) at 30 min before ischemia (early CaEDTA) or at 48 60 hr (late CaEDTA), but not 3-6 hr, after ischemia, afforded robust protection of CA1 neurons in approximately 50% (late CaEDTA) to 75% (early CaEDTA) of animals. We also show that Zn2+ acts via temporally distinct mechanisms to promote neuronal death. Early CaEDTA attenuated ischemia-induced GluR2 mRNA and protein downregulation (and, by inference, formation of Zn2+-permeable AMPARs), the delayed rise in Zn2+, and neuronal death. These findings suggest that Zn2+ acts at step(s) upstream from GluR2 gene downregulation and implicate Zn2+ in transcriptional regulation and/or GluR2 mRNA stability. Early CaEDTA also blocked mitochondrial release of cytochrome c and Smac/DIABLO (second mitochondria derived activator of caspases/direct inhibitor of apoptosis protein-binding protein with low pI), caspase-3 activity (but not procaspase-3 cleavage), p75NTR induction, and DNA fragmentation. These findings indicate that CaEDTA preserves the functional integrity of the mitochondrial outer membrane and arrests the caspase death cascade. Late injection of CaEDTA at a time when GluR2 is downregulated and caspase is activated inhibited the delayed rise in Zn2+, p75NTR induction, DNA fragmentation, and cell death. The finding of neuroprotection by late CaEDTA administration has striking implications for intervention in the delayed neuronal death associated with global ischemia. PMID- 15525777 TI - HCN2 and HCN1 channels govern the regularity of autonomous pacemaking and synaptic resetting in globus pallidus neurons. AB - The globus pallidus (GP) is a critical component of the basal ganglia circuitry controlling motor behavior. Dysregulation of GP activity has been implicated in a number of psychomotor disorders, including Parkinson's disease (PD), in which a cardinal feature of the pathophysiology is an alteration in the pattern and synchrony of discharge in GP neurons. Yet the determinants of this activity in GP neurons are poorly understood. To help fill this gap, electrophysiological, molecular, and computational approaches were used to identify and characterize GABAergic GP neurons in tissue slices from rodents. In vitro, GABAergic GP neurons generate a regular, autonomous, single-spike pacemaker activity. Hyperpolarization-activated, cyclic nucleotide-gated cation (HCN) channels make an important contribution to this process: their blockade with ZD7288 significantly slowed discharge rate and decreased its regularity. HCN currents evoked by somatic voltage clamp had fast and slow components. Single-cell RT-PCR and immunohistochemical approaches revealed robust expression of HCN2 subunits as well as significant levels of HCN1 subunits in GABAergic GP neurons. Transient activation of striatal GABAergic input to GP neurons led to a resetting of rhythmic discharge that was dependent on HCN currents. Simulations suggested that the ability of transient striatal GABAergic input to reset pacemaking was dependent on dendritic HCN2/HCN1 channels. Together, these studies show that HCN channels in GABAergic GP neurons are key determinants of the regularity and rate of pacemaking as well as striatal resetting of this activity, implicating HCN channels in the emergence of synchrony in PD. PMID- 15525778 TI - Target-dependent release of a presynaptic neuropeptide regulates the formation and maturation of specific synapses in Aplysia. AB - The correct wiring of neurons is critical for the normal functioning of the nervous system. Sensory neurons of Aplysia form synapses with specific postsynaptic targets. Interaction with appropriate target cells in culture induces a significant increase in axon growth, the number of sensory neuron varicosities with release sites contacting the target, and regulates the expression and distribution of mRNAs encoding presynaptic proteins such as syntaxin and the sensory neuron-specific neuropeptide sensorin. Synapse stabilization is accompanied by the maintenance of presynaptic varicosities and target-dependent regulation of mRNA distributions. We report here that specific targets induce the release of sensorin from sensory neurons, which then regulates synaptic efficacy, axonal growth associated with synapse formation, the maintenance of synaptic contacts, and the specific distribution of mRNAs. Bath application of an antisensorin antibody during the early phase of synapse formation blocked the expected increase in synaptic strength, the growth and formation of new presynaptic varicosities, and the target-dependent regulation of mRNA distribution. In contrast, bath application of sensorin accelerated the increase in synaptic strength and enhanced the formation of new varicosities and target-dependent regulation of mRNA distribution in sensory neurons. As synapses stabilize, sensorin secretion declines but is required for the maintenance of synaptic efficacy, presynaptic varicosities, and mRNA distributions. These results suggest that a retrograde target signal regulates the secretion and actions of a presynaptic neuropeptide critical for the formation and maintenance of specific synapses. PMID- 15525779 TI - Macrophage migration inhibitory factor: an intracellular inhibitor of angiotensin II-induced increases in neuronal activity. AB - Angiotensin II (Ang II) elicits Ang II type 1 receptor (AT1-R)-mediated increases in neuronal firing within the hypothalamus and brainstem that are ultimately responsible for physiological actions such as increased blood pressure and fluid intake. Although there is a growing literature on the intracellular mechanisms that mediate the actions of Ang II via AT1-R in neurons, little is known about the mechanisms that diminish or "switch-off" the neuronal chronotropic action of Ang II. In the present study, we identified macrophage migration inhibitory factor (MIF) as an intracellular inhibitor of the actions of Ang II in neurons. The evidence is as follows. First, Ang II, acting via AT1-R, increases the intracellular levels of MIF in neurons cultured from rat hypothalamus and brainstem. Second, elevation of intracellular MIF by Ang II prevents further chronotropic actions of this peptide. Third, intracellular application of exogenous recombinant MIF abolishes the Ang II-induced chronotropic action in neurons. Finally, intracellular application of the MIF peptide fragment MIF-(50 65), which harbors the thiol oxidoreductase property of the MIF molecule, mimics the inhibitory actions of MIF on Ang II-stimulated neuronal firing. Thus, this study is the first to demonstrate the existence of an intracellular negative regulator of Ang II-induced actions in neurons and indicates that MIF may act as a physiological brake for the chronotropic effects of Ang II in rat neurons. PMID- 15525780 TI - Circuitry for associative plasticity in the amygdala involves endocannabinoid signaling. AB - Endocannabinoids are crucial for the extinction of aversive memories, a process that considerably involves the amygdala. Here, we show that low-frequency stimulation of afferents in the lateral amygdala with 100 pulses at 1 Hz releases endocannabinoids postsynaptically from neurons of the basolateral amygdala of mice in vitro and thereby induces a long-term depression of inhibitory GABAergic synaptic transmission (LTDi) via a presynaptic mechanism. Lowering inhibitory synaptic transmission significantly increases the amplitude of excitatory synaptic currents in principal neurons of the central nucleus, which is the main output site of the amygdala. LTDi involves a selective mGluR1 (metabotropic glutamate receptor 1)-mediated calcium-independent mechanism and the activation of the adenylyl cyclase-protein kinase A pathway. LTDi is abolished by the cannabinoid type 1 (CB1) receptor antagonist SR141716A and cannot be evoked in CB1 receptor-deficient animals. LTDi is significantly enhanced in mice lacking the anandamide-degrading enzyme fatty acid amide hydrolase. The present findings show for the first time that mGluR activation induces a retrograde endocannabinoid signaling via activation of the adenylyl cyclase-protein kinase A pathway and the release of anandamide. Furthermore, the results indicate that anandamide decreases the activity of inhibitory interneurons in the amygdala. This disinhibition increases the activity of common output neurons and could provide a prerequisite for extinction by formation of new memory. PMID- 15525781 TI - Molecular organization of the ferret visual thalamus. AB - The visual system encodes and deciphers information using parallel, anatomically segregated, processing streams. To reveal patterns of gene expression in the visual thalamus correlated with physiological processing streams, we designed a custom ferret cDNA microarray. By isolating specific subregions and layers of the thalamus, we identified a set of transcription factors, including Zic2, Islet1, and Six3, the unique distribution profiles of which differentiated the lateral geniculate nucleus (LGN) from the associated perigeniculate nucleus. Within the LGN, odd homeobox1 differentiated the A layers, which contain X cells and Y cells, from the C layers. One neuron-specific protein, Purkinje cell protein 4 (PCP4), was strongly expressed in Y cells in the ferret LGN and in the magnocellular layers of the primate LGN. In the ferret LGN, PCP4 expression began as early as postnatal day 7 (P7), suggesting that Y cells are already specified by P7. These results reveal a rich molecular repertoire that correlates with functional divisions of the LGN. PMID- 15525782 TI - Stimulation of the posterior parietal cortex interferes with arm trajectory adjustments during the learning of new dynamics. AB - Substantial neurophysiological evidence points to the posterior parietal cortex (PPC) as playing a key role in the coordinate transformation necessary for visually guided reaching. Our goal was to examine the role of PPC in the context of learning new dynamics of arm movements. We assessed this possibility by stimulating PPC with transcranial magnetic stimulation (TMS) while subjects learned to make reaching movements with their right hand in a velocity-dependent force field. We reasoned that, if PPC is necessary to adjust the trajectory of the arm as it interacts with a novel mechanical system, interfering with the functioning of PPC would impair adaptation. Single pulses of TMS were applied over the left PPC 40 msec after the onset of movement during adaptation. As a control, another group of subjects was stimulated over the visual cortex. During early stages of learning, the magnitude of the error (measured as the deviation of the hand paths) was similar across groups. By the end of the learning period, however, error magnitudes decreased to baseline levels for controls but remained significantly larger for the group stimulated over PPC. Our findings are consistent with a role of PPC in the adjustment of motor commands necessary for adapting to a novel mechanical environment. PMID- 15525783 TI - Caspase-7 expanded function and intrinsic expression level underlies strain specific brain phenotype of caspase-3-null mice. AB - Caspase-3-deficient mice of the 129S1/SvImJ (129) strain show severe brain development defects resulting in brain overgrowth and perinatal lethality, whereas on the C57BL/6J (B6) background, these mice develop normally. We therefore sought to identify the strain-dependent ameliorating gene. We biochemically isolated caspase-7 from B6-caspase-3-null (Casp3-/-) tissues as being the enzyme with caspase-3-like properties and capability of performing a caspase-3 surrogate function, apoptotic DNA fragmentation. Moreover, we show that, in contrast to the human enzymes, mouse caspase-7 is as efficient as caspase-3 at cleaving and thus inactivating ICAD (inhibitor of caspase-activated DNase), the inhibitor of apoptotic DNA fragmentation. Low levels of caspase-7 expression and activation correlate with lack of DNA fragmentation in 129-Casp3-/ apoptotic precursor neurons, whereas B6-Casp3-/- cells, which can fragment their DNA, show higher levels of caspase-7 expression and activation. The amount of caspase-7 activation in apoptotic precursor neurons is independent of the presence of caspase-3. Together, our findings demonstrate for the first time a strong correlation between caspase-7 activity, normal brain development, and apoptotic DNA fragmentation in Casp3-/- mice. PMID- 15525784 TI - Transcranial direct current stimulation during sleep improves declarative memory. AB - In humans, weak transcranial direct current stimulation (tDCS) modulates excitability in the motor, visual, and prefrontal cortex. Periods rich in slow wave sleep (SWS) not only facilitate the consolidation of declarative memories, but in humans, SWS is also accompanied by a pronounced endogenous transcortical DC potential shift of negative polarity over frontocortical areas. To experimentally induce widespread extracellular negative DC potentials, we applied anodal tDCS (0.26 mA) [correction] repeatedly (over 30 min) bilaterally at frontocortical electrode sites during a retention period rich in SWS. Retention of declarative memories (word pairs) and also nondeclarative memories (mirror tracing skills) learned previously was tested after this period and compared with retention performance after placebo stimulation as well as after retention intervals of wakefulness. Compared with placebo stimulation, anodal tDCS during SWS-rich sleep distinctly increased the retention of word pairs (p < 0.005). When applied during the wake retention interval, tDCS did not affect declarative memory. Procedural memory was also not affected by tDCS. Mood was improved both after tDCS during sleep and during wake intervals. tDCS increased sleep depth toward the end of the stimulation period, whereas the average power in the faster frequency bands (,alpha, and beta) was reduced. Acutely, anodal tDCS increased slow oscillatory activity <3 Hz. We conclude that effects of tDCS involve enhanced generation of slow oscillatory EEG activity considered to facilitate processes of neuronal plasticity. Shifts in extracellular ionic concentration in frontocortical tissue (expressed as negative DC potentials during SWS) may facilitate sleep-dependent consolidation of declarative memories. PMID- 15525785 TI - Glycogen synthase kinase-3beta phosphorylates Bax and promotes its mitochondrial localization during neuronal apoptosis. AB - Glycogen synthase kinase-3beta (GSK-3beta) is a critical activator of neuronal apoptosis induced by a diverse array of neurotoxic insults. However, the downstream substrates of GSK-3beta that ultimately induce neuronal death are unknown. Here, we show that GSK-3beta phosphorylates and regulates the activity of Bax, a pro-apoptotic Bcl-2 family member that stimulates the intrinsic (mitochondrial) death pathway by eliciting cytochrome c release from mitochondria. In cerebellar granule neurons undergoing apoptosis, inhibition of GSK-3beta suppressed both the mitochondrial translocation of an expressed green fluorescent protein (GFP)-Bax(alpha) fusion protein and the conformational activation of endogenous Bax. GSK-3beta directly phosphorylated Bax(alpha) on Ser163, a residue found within a species-conserved, putative GSK-3beta phosphorylation motif. Coexpression of GFP-Bax(alpha) with a constitutively active mutant of GSK-3beta, GSK-3beta(Ser9Ala), enhanced the in vivo phosphorylation of wild-type Bax(alpha), but not a Ser163Ala mutant of Bax(alpha), in transfected human embryonic kidney 293 (HEK293) cells. Moreover, cotransfection with constitutively active GSK-3beta promoted the localization of Bax(alpha) to mitochondria and induced apoptosis in both transfected HEK293 cells and cerebellar granule neurons. In contrast, neither a Ser163Ala point mutant of Bax(alpha) nor a naturally occurring splice variant that lacks 13 amino acids encompassing Ser163 (Bax(sigma)) were driven to mitochondria in HEK293 cells coexpressing constitutively active GSK-3beta. In a similar manner, either mutation or deletion of the identified GSK-3beta phosphorylation motif prevented the localization of Bax to mitochondria in cerebellar granule neurons undergoing apoptosis. Our results indicate that GSK-3beta exerts some of its pro-apoptotic effects in neurons by regulating the mitochondrial localization of Bax, a key component of the intrinsic apoptotic cascade. PMID- 15525786 TI - p53 activation domain 1 is essential for PUMA upregulation and p53-mediated neuronal cell death. AB - The p53 tumor suppressor gene has been implicated in the regulation of apoptosis in a number of different neuronal death paradigms. Because of the importance of p53 in neuronal injury, we questioned the mechanism underlying p53-mediated apoptosis in neurons. Using adenoviral-mediated gene delivery, reconstitution experiments, and mice carrying a knock-in mutation in the endogenous p53 gene, we show that the transactivation function of p53 is essential to induce neuronal cell death. Although p53 possesses two transactivation domains that can activate p53 targets independently, we demonstrate that the first activation domain (ADI) is required to drive apoptosis after neuronal injury. Furthermore, the BH3-only proteins Noxa and PUMA exhibit differential regulation by the two transactivation domains. Here, we show that Noxa can be induced by either activation domain, whereas PUMA induction requires both activation domains to be intact. Unlike Noxa, the upregulation of PUMA alone is sufficient to induce neuronal cell death. We demonstrate, therefore, that the first transactivation domain of p53 is indispensable for the induction of neuronal cell death. PMID- 15525787 TI - Endocannabinoids link feeding state and auditory perception-related gene expression. AB - Singing by adult male zebra finches is a learned behavior important for courtship, kin recognition, and nest defense (Zann, 1996) and is inhibited by both brief periods of limited food availability and systemic injection of cannabinoids. These similar effects on singing, combined with increasing evidence for endocannabinoid involvement in feeding behavior, led us to evaluate a possible shared mechanism. We found that limited food availability both reduces singing in a cannabinoid antagonist-reversible manner and increases levels of the endocannabinoid 2-arachidonyl glycerol in various brain regions including the caudal telencephalon, an area that contains auditory telencephalon including the L2 subfield of L (L2) and caudal medial nidopallium (NCM). Development and use of an anti-zebra finch cannabinoid receptor type 1 (CB1) antibody demonstrates distinct, dense cannabinoid receptor expression within song regions including Area X, lMAN (lateral magnocellular nucleus of anterior nidopallium), HVC, RA (robust nucleus of arcopallium), and L2. NCM receives L2 projections and is implicated in integration of auditory information. Activity in this area, determined through expression of the transcription factor ZENK, is increased after exposure to unfamiliar song. Because previous work has shown that these novel song-stimulated increases in NCM activity are mitigated by cannabinoid exposure, we tested and found that similar effects on ZENK expression are produced by limiting food. Limited food-related reductions in the activity of NCM neurons were reversed by the cannabinoid antagonist SR141716A (N-piperidino-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3-carboxamide), implicating CB1 cannabinoid receptor involvement. Taken together, these experiments indicate a link between feeding state and gene expression related to auditory perception that is mediated by endocannabinoid signaling. PMID- 15525788 TI - A novel epilepsy mutation in the sodium channel SCN1A identifies a cytoplasmic domain for beta subunit interaction. AB - A mutation in the sodium channel SCN1A was identified in a small Italian family with dominantly inherited generalized epilepsy with febrile seizures plus (GEFS+). The mutation, D1866Y, alters an evolutionarily conserved aspartate residue in the C-terminal cytoplasmic domain of the sodium channel alpha subunit. The mutation decreased modulation of the alpha subunit by beta1, which normally causes a negative shift in the voltage dependence of inactivation in oocytes. There was less of a shift with the mutant channel, resulting in a 10 mV difference between the wild-type and mutant channels in the presence of beta1. This shift increased the magnitude of the window current, which resulted in more persistent current during a voltage ramp. Computational analysis suggests that neurons expressing the mutant channels will fire an action potential with a shorter onset delay in response to a threshold current injection, and that they will fire multiple action potentials with a shorter interspike interval at a higher input stimulus. These results suggest a causal relationship between a positive shift in the voltage dependence of sodium channel inactivation and spontaneous seizure activity. Direct interaction between the cytoplasmic C terminal domain of the wild-type alpha subunit with the beta1 or beta3 subunit was first demonstrated by yeast two-hybrid analysis. The SCN1A peptide K1846 R1886 is sufficient for beta subunit interaction. Coimmunoprecipitation from transfected mammalian cells confirmed the interaction between the C-terminal domains of the alpha and beta1 subunits. The D1866Y mutation weakens this interaction, demonstrating a novel molecular mechanism leading to seizure susceptibility. PMID- 15525789 TI - The role of polymorphisms of glutathione S-transferases GSTM1, M3, P1, T1 and A1 in susceptibility to alcoholic liver disease. AB - AIMS AND METHODS: Oxidant stress is proposed to be an important pathogenic factor in liver damage related to alcohol. The glutathione S-transferases (GSTs) are a group of polymorphic enzymes that are important in protection against oxidant stress. As there is evidence for genetic susceptibility to alcohol-related liver disease we have compared the frequency of polymorphisms of GSTM1, M3, P1, T1 and A1 by polymerase chain reaction (PCR) on leucocyte DNA in patients from North Staffordshire, Birmingham and Liverpool with alcohol-related chronic liver disease heavy drinking and normal local controls. RESULTS: There were no significant differences in GSTM1, GSTM3 or GSTP1 genotype frequencies among patients, drinking and non-drinking controls from the three centres. There was a significant increase in the GSTT1 null Liverpool alcoholic liver disease (ALD) patients compared with corresponding non-drinking controls (26.3 and 14.6%, respectively; P = 0.044, odds ratio (OR) = 2.1, 95% CI = 1.1-4.7) though this was not repeated in the Birmingham and North Staffordshire cohorts. For GSTA1, the 69 CC genotype was associated with increased risk of ALD in the Liverpool group, but a reduced risk in the North Staffordshire group. CONCLUSIONS: We have failed to demonstrate within the limitation of a case-control study a reproducible significant association of GST polymorphisms with susceptibility to ALD but there are suggestions that GSTA1 and GSTT1 warrant further study. PMID- 15525790 TI - A one-year pragmatic trial of naltrexone vs disulfiram in the treatment of alcohol dependence. AB - AIMS: To compare the efficacy of naltrexone and disulfiram in preventing an alcoholic relapse in routine clinical practice in an Indian metropolis. METHODS: Hundred alcohol-dependent men, for whom a family member would accompany the patient to follow-up appointments, were randomly allocated to a year of treatment with either naltrexone or disulfiram. Patients, the accompanying family member and the treating psychiatrist were aware of the nature of treatment given. Alcohol consumption, craving and adverse events were recorded weekly for the first three months, then fortnightly for the rest of the year, by the treating psychiatrist. Serum gamma-glutamyl transferase (GGT) was measured at the start and the end of the study. RESULTS: At the end of the year, 97 patients were still in contact. Relapse, the consumption of >5 drinks (40 g of ethanol) in a 24 h period, occurred at a mean of 119 days with disulfiram and at 63 days with naltrexone (P = 0.020). Mean serum GGT, which had not differed between the two groups initially, was 117 U/l with naltrexone and 85 U/l with disulfiram (P = 0.038) at the end of the study. Eighty-six per cent of the patients remained abstinent throughout the study with disulfiram compared to 44% with naltrexone (P = 0.0009). However, patients allocated to naltrexone had significantly lower craving than those allocated to disulfiram. CONCLUSIONS: Disulfiram is superior to naltrexone in preventing a relapse among alcohol-dependent men with family support. Comparison between these treatments in other settings and in different types of alcoholics is warranted. PMID- 15525791 TI - Ethyl pyruvate inhibits nuclear factor-kappaB-dependent signaling by directly targeting p65. AB - Ethyl pyruvate has been shown to have anti-inflammatory properties in numerous cell culture and animal studies. In this series of experiments, we tested the hypothesis that ethyl pyruvate inhibits signaling by the pro-inflammatory transcription factor, NF-kappaB. Ethyl pyruvate inhibited luciferase expression in lipopolysaccharide-stimulated murine macrophage-like RAW 264.7 cells transfected with an NF-kappaB-dependent luciferase reporter vector. Ethyl pyruvate also decreased NF-kappaB DNA-binding activity in lipopolysaccharide stimulated RAW 264.7 cells and decreased lipopolysaccharide-induced expression of an NF-kappaB-dependent gene, inducible nitric oxide synthase. Ethyl pyruvate had no effect on the degradation of IkappaBalpha or IkappaBbeta in lipopolysaccharide stimulated RAW 264.7 cells, suggesting that ethyl pyruvate acts distally to this step in the activation of NF-kappaB. In a cell-free system, binding of p50 homodimers to an NF-kappaB consensus oligonucleotide sequence was unaffected by ethyl pyruvate over a wide range of concentrations, indicating that ethyl pyruvate probably does not modify or interact with the p50 subunit of NF-kappaB. In contrast, ethyl pyruvate inhibited DNA binding by ectopically overexpressed wild-type p65 homodimers. However, ethyl pyruvate failed to inhibit the DNA binding activity of homodimers of an overexpressed mutant form of a p65 with substitution of serine for cysteine 38. Taken together, these results suggest that ethyl pyruvate inhibits DNA-binding by covalently modifying p65 at Cys(38). We conclude that some of the beneficial anti-inflammatory effects of ethyl pyruvate may be due to modification of p65, thereby inhibiting signaling via the NF-kappaB pathway. PMID- 15525792 TI - A new ATP-sensitive potassium channel opener reduces blood pressure and reverses cardiovascular remodeling in experimental hypertension. AB - Some potassium channel openers (KCOs) are potent vasodilators that mainly target the ATP-sensitive potassium channels in vascular smooth muscle cells. Their lack of tissue selectivity limits their clinical use in hypertension therapy. Iptakalim [2,3-dimethyl-n-(1-methylethyl)-2-butylamine], which belongs to a novel chemical type of KCO, possesses unique pharmacological characteristics. In vitro experiments have shown that iptakalim could limit its vasorelaxing actions to resistance vessels. In this study, we investigate the antihypertensive effects of iptakalim on two different experimental hypertensive models: stroke-prone, spontaneously hypertensive rats (SHRsps) and two-kidney with one-clip renal hypertensive dogs (2K1C RHD). In acute hypotensive tests, iptakalim showed stable, long-lasting antihypertensive effects in SHRsps and 2K1C RHDs. Mean while, it had little effect on heart rate when compared with pinacidil, nifedipine, captopril, or bisoprolol. In experimental therapeutic tests, repeated doses in SHRsps for 30 days or in 2K1C RHDs for 14 days produced consistent antihypertensive effects without causing tolerance. In separate experiments, chronic administration of iptakalim resulted in reversing hypertensive vascular remodeling in spontaneously hypertensive rats and hypertensive cardiac remodeling in SHRsps. These results suggest that iptakalim is a promising antihypertensive drug. PMID- 15525793 TI - Cilostazol prevents remnant lipoprotein particle-induced monocyte adhesion to endothelial cells by suppression of adhesion molecules and monocyte chemoattractant protein-1 expression via lectin-like receptor for oxidized low density lipoprotein receptor activation. AB - This study shows cilostazol effect to prevent remnant lipoprotein particle (RLP) induced monocyte adhesion to human umbilical vein endothelial cells (HUVECs). Upon incubation of HUVECs with RLP (50 microg/ml), adherent monocytes significantly increased by 3.3-fold with increased cell surface expression of vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1, E selectin, and monocyte chemoattractant protein-1 (MCP-1). Cilostazol ( approximately 1-100 microM) concentration dependently repressed these variables as did (E)3-[(4-t-butylphenyl)sulfonyl]-2-propenenitrile (BAY 11-7085) (10 microM), a specific nuclear factor-kappaB (NF-kappaB) inhibitor. Cilostazol effects were significantly antagonized by iberiotoxin (1 microM), a maxi-K channel blocker. RLP significantly increased expression of lectin-like receptor for oxidized low-density lipoprotein (LDL) (LOX-1) receptor protein. Upon transfection with antisense LOX-1 oligodeoxynucleotide (As-LOX-1), LOX-1 receptor expression was reduced, whereas HUVECs with sense LOX-1 oligodeoxynucleotide did express high LOX-1 receptor. RLP-stimulated superoxide and tumor necrosis factor alpha levels were significantly lowered with decreased expression of VCAM-1 and MCP-1 by transfection with As-LOX-1 as did polyinosinic acid (10 microg/ml, a LOX 1 receptor inhibitor). RLP significantly degraded inhibitory kappaBalpha in the cytoplasm and activated nuclear factor-kappaB (NF-kappaB) p65 in the nucleus of HUVECs with increased luciferase activity of NF-kappaB, all of which were reversed by cilostazol (10 microM), BAY 11-7085, and polyinosinic acid. Together, cilostazol suppresses RLP-stimulated increased monocyte adhesion to HUVECs by suppression of LOX-1 receptor-coupled NF-kappaB-dependent nuclear transcription via mediation of the maxi-K channel opening. PMID- 15525794 TI - Mixed-lineage kinase inhibitors require the activation of Trk receptors to maintain long-term neuronal trophism and survival. AB - Small-molecule mixed-lineage kinase (MLK) inhibitors, such as CEP-1347 [3,9 bis[(ethylthio)methyl]-(8R*,9S*,11S*)-(-)-9-hydroxy-9-methoxycarbonyl-8-methyl 2,3,9,10-tetrahydro-8,11-epoxy-1H,8H, 11H-2,7b,11a triazadibenzo(a,g)cycloocta(cde)trinden-1-one] and CEP-11004 [3,9-bis [(isopropylthio)methyl]-(8R*,9S*,11S*)-(-)-9-hydroxy-9-methoxycarbonyl-8-methyl 2,3,9,10-tetrahydro-8,11-epoxy-1H,8H,11H-2,7b,11a triazadibenzo(a,g)cycloocta(cde)trinden-1-one], prevent c-Jun NH(2)-terminal kinase (JNK) pathway activation as well as the consequent neuronal cell death in many cell culture and animal models. In the cell culture model of nerve growth factor (NGF)-deprived sympathetic neurons, we find that CEP-11004 induced a approximately 3-fold increase in the mRNA and protein levels of TrkA, the NGF receptor. This resulted in ligand-independent activation of the TrkA receptor and the downstream phosphatidylinositol 3-kinase (PI3-kinase) pathway. Addition of the Trk inhibitor K252a [(8R*,9S*,11S*)-(-)-9-hydroxy-9-methoxycarbonyl-8-methyl 2,3,9,10-tetrahydro-8,11-epoxy-1H,8H,11H-2,7b,11a triazadibenzo(a,g)cycloocta(cde)-trinden-1-one] or the PI3-kinase inhibitor LY294002 [2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one] significantly decreased the protein synthesis rates, mitochondrial function, and neuronal survival maintained by CEP-11004. In contrast to sympathetic neurons, MLK inhibitors maintain only short-term survival of potassium- and serum-deprived rat cerebellar granule neurons (CGNs), despite continuous inhibition of the JNK pathway. We found that similar to sympathetic neurons, CEP-11004 increased the levels of the Trk receptor expressed in CGNs, TrkB. However, CGNs required the addition of the exogenous ligand brain-derived neurotrophic factor (BDNF) to activate the PI3-kinase pathway and to maintain long-term survival. BDNF activated TrkB, but caused rapid down-regulation of activated receptors and maintained only minimal survival. Therefore, increase in TrkB levels by CEP-11004 mediated a synergism with BDNF resulting in long-term survival in response to the combined treatment of CEP-11004 and BDNF. Taken together, our studies suggest that in addition to the direct inhibition of the JNK pathway, the indirect activation of the PI3-kinase pathway via Trk activation is important for MLK inhibitor-mediated neuronal survival and trophism. PMID- 15525795 TI - YC-1 [3-(5'-Hydroxymethyl-2'-furyl)-1-benzyl Indazole] exhibits a novel antiproliferative effect and arrests the cell cycle in G0-G1 in human hepatocellular carcinoma cells. AB - This study delineates the antiproliferative activities and in vivo efficacy of YC 1 [3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole] in human hepatocellular carcinoma cells. YC-1 inhibited the growth of HA22T and Hep3B cells in a concentration-dependent manner without significant cytotoxicity. YC-1 induced G(1) phase arrest in the cell cycle, as detected by an increase in the proportion of cells in the G(1) phase using FAC-Scan flow cytometric analysis. It was further shown that cGMP, p42/p44 mitogen-activated protein kinase, or AKT kinase mediated signaling pathways did not contribute to the YC-1-induced effect. Of note, YC-1 induced a dramatic increase in the expression of cyclin-dependent kinase (CDK)-inhibitory protein, p21(CIP1/WAP1), and a modest increase in p27(KIP1). The association of p21(CIP1/WAP1) with CDK2 was markedly increased in cells responsive to YC-1. YC-1 did not modify the expression of cyclin D1, cyclin E, CDK2, or CDK4. In a corollary in vivo study, YC-1 induced dose-dependent inhibition of tumor growth in mice inoculated with HA22T cells. Immunohistochemical analysis revealed an inverse relationship between the staining of p21(CIP1/WAF) and the staining of Ki-67, a cell proliferation marker. Based on the results reported herein, we suggest that YC-1 induces cell cycle arrest and inhibits tumor growth both in vitro and in vivo via the up-regulation of p21(CIP1/WAP1) expression in HA22T cells. Because of this, YC-1 is a potential antitumor agent worthy of further investigation. PMID- 15525796 TI - Immunogenicity and rapid blood clearance of liposomes containing polyethylene glycol-lipid conjugates and nucleic Acid. AB - Polyethylene glycol (PEG) is used widely in the pharmaceutical industry to improve the pharmacokinetics and reduce the immunogenicity of therapeutic and diagnostic agents. The incorporation of lipid-conjugated PEG into liposomal drug delivery systems greatly enhances the circulation times of liposomes by providing a protective, steric barrier against interactions with plasma proteins and cells. Here we report that liposome compositions containing PEG-lipid derivatives and encapsulated antisense oligodeoxynucleotide (ODN) or plasmid DNA elicit a strong immune response that results in the rapid blood clearance of subsequent doses in mice. The magnitude of this response is sufficient to induce significant morbidity and, in some instances, mortality. This effect has been observed in several strains of mice and was independent of sequence motifs, such as immunostimulatory CpG motifs. The ODN-to-lipid ratio and ODN dose was also determined to be important, with abrogation of the response occurring at a ratio between 0.04 and 0.08 (w/w). Rapid elimination of liposome-encapsulated ODN from blood depends on the presence of PEG-lipid in the membrane because the use of nonpegylated liposomes or liposomes containing rapidly exchangeable PEG-lipid also abrogated the response. These studies have important implications for the evaluation and therapeutic use of liposomal formulations of nucleic acid, as well as the potential development of liposomal vaccines. PMID- 15525797 TI - Cannabinoid CB1 receptor antagonists as promising new medications for drug dependence. AB - This review examines the development of cannabinoid CB(1) receptor antagonists as a new class of therapeutic agents for drug addiction. Abused drugs [alcohol, opiates, Delta(9)-tetrahydrocannabinol (Delta(9)-THC), and psychostimulants, including nicotine] elicit a variety of chronically relapsing disorders by interacting with endogenous neural pathways in the brain. In particular, they share the common property of activating mesolimbic dopamine brain reward systems, and virtually all abused drugs elevate dopamine levels in the nucleus accumbens. Cannabinoid CB(1) receptors are expressed in this brain reward circuit and modulate the dopamine-releasing effects of Delta(9)-THC and nicotine. Rimonabant (SR141716), a CB(1) receptor antagonist, blocks both the dopamine-releasing and discriminative and rewarding effects of Delta(9)-THC in animals. Blockade of CB(1) receptor activity by genetic invalidation also decreases rewarding effects of opiates and alcohol in animals. Although CB(1) receptor blockade is generally ineffective in reducing the self-administration of cocaine in rodents and primates, it reduces the reinstatement of extinguished cocaine-seeking behavior produced by cocaine-associated conditioned stimuli and cocaine-priming injections. Likewise, CB(1) receptor blockade is effective in reducing nicotine seeking behavior induced by re-exposure to nicotine-associated stimuli. Some of these findings have been recently validated in humans. In clinical trials, Rimonabant blocks the subjective effects of Delta(9)-THC in humans and prevents relapse to smoking in exsmokers. Findings from both clinical and preclinical studies suggest that ligands blocking CB(1) receptors offer a novel approach for patients suffering from drug dependence that may be efficacious across different classes of abused drugs. PMID- 15525798 TI - Angiotensin II-induced Akt activation through the epidermal growth factor receptor in vascular smooth muscle cells is mediated by phospholipid metabolites derived by activation of phospholipase D. AB - Angiotensin II (Ang II) activates cytosolic Ca(2+)-dependent phospholipase A(2) (cPLA(2)), phospholipase D (PLD), p38 mitogen-activated protein kinase (MAPK), epidermal growth factor receptor (EGFR) and Akt in vascular smooth muscle cells (VSMC). This study was conducted to investigate the relationship between Akt activation by Ang II and other signaling molecules in rat VSMC. Ang II-induced Akt phosphorylation was significantly reduced by the PLD inhibitor 1-butanol, but not by its inactive analog 2-butanol, and by brefeldin A, an inhibitor of the PLD cofactor ADP-ribosylation factor, and in cells infected with retrovirus containing PLD(2) siRNA or transfected with PLD(2) antisense but not control LacZ or sense oligonucleotide. Diacylglycerol kinase inhibitor II diminished Ang II induced and diC8-phosphatidic acid (PA)-increased Akt phosphorylation, suggesting that PLD-dependent Akt activation is mediated by PA. Ang II-induced EGFR phosphorylation was inhibited by 1-butanol and PLD(2) siRNA and also by cPLA(2) siRNA. In addition, the inhibitor of arachidonic acid (AA) metabolism 5,8,11,14 eicosatetraynoic acid (ETYA) reduced both Ang II- and AA-induced EGFR transactivation. Furthermore, ETYA, cPLA(2) antisense, and cPLA(2) siRNA attenuated Ang II-elicited PLD activation. p38 MAPK inhibitor SB202190 [4-(4 flurophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)1H-imidazole] reduced PLD activity and EGFR and Akt phosphorylation elicited by Ang II. Pyrrolidine-1, a cPLA(2) inhibitor, and cPLA(2) siRNA decreased p38 MAPK activity. These data indicate that Ang II-stimulated Akt activity is mediated by cPLA(2)-dependent, p38 MAPK regulated PLD(2) activation and EGFR transactivation. We propose the following scheme of the sequence of events leading to activation of Akt in VSMC by Ang II: Ang II-->cPLA(2)-->AA-->p38 MAPK-->PLD(2)-->PA-->EGFR-->Akt. PMID- 15525799 TI - Blood clearance and activity of erythrocyte-coupled fibrinolytics. AB - Conjugating tissue-type plasminogen activator (tPA) to red blood cells (RBCs) endows it with features useful for thromboprophylaxis. However, the optimal intensity and duration of thromboprophylaxis vary among clinical settings. To assess how the intrinsic properties of a plasminogen activator (PA) affect functions of the corresponding RBC/PA conjugate, we coupled equal amounts of tPA or Retavase (rPA; a variant with an extended circulation time, lower fibrin affinity, and greater susceptibility to PA inhibitors). Conjugation to RBC markedly prolonged the circulation of each PA in rats and mice, without detrimental effects on carrier RBC. The initial blood clearance of RBC/tPA was faster than RBC/rPA, yet it exerted greater fibrinolytic activity, in part due to greater resistance of tPA and RBC/tPA to plasma inhibitors versus rPA and RBC/rPA observed in vitro. Soluble and RBC-coupled tPA and rPA exerted the same amidolytic activity, yet RBC/tPA lysed fibrin clots more effectively than RBC/rPA, notwithstanding comparable fibrinolytic activity of their soluble counterparts. Conjugation to RBC suppressed rPA's ability to be activated by fibrin, whereas the fibrin activation of RBC-coupled tPA was not hindered. Therefore, the functional profile of RBC/PA is influenced by: pharmacokinetic features provided by carrier RBC (e.g., prolonged circulation), intrinsic PA features (e.g., clearance rate, resistance to inhibitors), and changes imposed by conjugation to RBC (e.g., loss of cofactor stimulation). These factors, different from those guiding the design of soluble PA for lysis of existing clots, can be exploited in the rational design of RBC/PA tailored for specific prophylactic indications. PMID- 15525800 TI - Pharmacological properties and functional role of a TRP-related ion channel in lobster olfactory receptor neurons. AB - Odors activate lobster olfactory receptor neurons (ORNs) through phosphoinositide signaling that appears to target a Na(+)-gated nonselective cation channel. The Na(+)-gated channel is a potential member of the growing family of transient receptor potential (TRP) channels. Here, we test the effect of potential antagonists on the channel in cell-free patches from cultured lobster ORNs. We show that the channel is antagonized by H+ and the TRP channel blockers 2 aminoethoxydiphenyl borate, SKF96365, ruthenium red, Al3+, Gd3+, and La3+. We then use this enhanced antagonist profile together with the agonists Na+ and Ca2+ to implicate the channel in signal amplification in the cells. PMID- 15525801 TI - Single-column thalamocortical network model exhibiting gamma oscillations, sleep spindles, and epileptogenic bursts. AB - To better understand population phenomena in thalamocortical neuronal ensembles, we have constructed a preliminary network model with 3,560 multicompartment neurons (containing soma, branching dendrites, and a portion of axon). Types of neurons included superficial pyramids (with regular spiking [RS] and fast rhythmic bursting [FRB] firing behaviors); RS spiny stellates; fast spiking (FS) interneurons, with basket-type and axoaxonic types of connectivity, and located in superficial and deep cortical layers; low threshold spiking (LTS) interneurons, which contacted principal cell dendrites; deep pyramids, which could have RS or intrinsic bursting (IB) firing behaviors, and endowed either with nontufted apical dendrites or with long tufted apical dendrites; thalamocortical relay (TCR) cells; and nucleus reticularis (nRT) cells. To the extent possible, both electrophysiology and synaptic connectivity were based on published data, although many arbitrary choices were necessary. In addition to synaptic connectivity (by AMPA/kainate, NMDA, and GABA(A) receptors), we also included electrical coupling between dendrites of interneurons, nRT cells, and TCR cells, and--in various combinations--electrical coupling between the proximal axons of certain cortical principal neurons. Our network model replicates several observed population phenomena, including 1) persistent gamma oscillations; 2) thalamocortical sleep spindles; 3) series of synchronized population bursts, resembling electrographic seizures; 4) isolated double population bursts with superimposed very fast oscillations (>100 Hz, "VFO"); 5) spike-wave, polyspike wave, and fast runs (about 10 Hz). We show that epileptiform bursts, including double and multiple bursts, containing VFO occur in rat auditory cortex in vitro, in the presence of kainate, when both GABA(A) and GABA(B) receptors are blocked. Electrical coupling between axons appears necessary (as reported previously) for persistent gamma and additionally plays a role in the detailed shaping of epileptogenic events. The degree of recurrent synaptic excitation between spiny stellate cells, and their tendency to fire throughout multiple bursts, also appears critical in shaping epileptogenic events. PMID- 15525802 TI - Synchronization in hybrid neuronal networks of the hippocampal formation. AB - Understanding the mechanistic bases of neuronal synchronization is a current challenge in quantitative neuroscience. We studied this problem in two putative cellular pacemakers of the mammalian hippocampal theta rhythm: glutamatergic stellate cells (SCs) of the medial entorhinal cortex and GABAergic oriens lacunosum-molecular (O-LM) interneurons of hippocampal region CA1. We used two experimental methods. First, we measured changes in spike timing induced by artificial synaptic inputs applied to individual neurons. We then measured responses of free-running hybrid neuronal networks, consisting of biological neurons coupled (via dynamic clamp) to biological or virtual counterparts. Results from the single-cell experiments predicted network behaviors well and are compatible with previous model-based predictions of how specific membrane mechanisms give rise to empirically measured synchronization behavior. Both cell types phase lock stably when connected via homogeneous excitatory-excitatory (E E) or inhibitory-inhibitory (I-I) connections. Phase-locked firing is consistently synchronous for either cell type with E-E connections and nearly anti-synchronous with I-I connections. With heterogeneous connections (e.g., excitatory-inhibitory, as might be expected if members of a given population had heterogeneous connections involving intermediate interneurons), networks often settled into phase locking that was either stable or unstable, depending on the order of firing of the two cells in the hybrid network. Our results imply that excitatory SCs, but not inhibitory O-LM interneurons, are capable of synchronizing in phase via monosynaptic mutual connections of the biologically appropriate polarity. Results are largely independent of synaptic strength and synaptic kinetics, implying that our conclusions are robust and largely unaffected by synaptic plasticity. PMID- 15525803 TI - Vestibular gaze stabilization: different behavioral strategies for arboreal and terrestrial avians. AB - In birds, it is thought that head movements play a major role in the reflexive stabilization of gaze and vision. In this study, we investigated the contributions of the eye and head to gaze stabilization during rotations under both head-fixed [vestibuloocular (VOR)] and head-free conditions in two avian species: pigeons and quails. These two species differ both in ocular anatomy (the pigeon has 2 distinct foveal regions), as well as in behavioral repertoires. Pigeons are arboreal, fly extended distances, and can navigate. Quails are primarily engrossed in terrestrial niches and fly only short distances. Unlike the head-fixed VOR gains that were under-compensatory for both species, gaze gains under head-free conditions were completely compensatory at high frequencies. This compensation was achieved primarily with head movements in pigeons, but with combined head and eye-in-head contributions in the quail. In contrast, eye-in-head motion, which was significantly reduced for head-free compared with head-fixed conditions, contributed very little to overall gaze stability in pigeons. These results suggest that disparity between the stabilization strategies employed by these two birds may be attributed to differences in species-specific behavior and anatomy. PMID- 15525804 TI - Skilled motor learning does not enhance long-term depression in the motor cortex in vivo. AB - Learning of motor skills may occur as a consequence of changes in the efficacy of synaptic connections in the primary motor cortex. We investigated if learning in a reaching task affects the excitability, short-term plasticity, and long-term plasticity of horizontal connections in layers II-III of the motor cortex. Because training in this task requires animals to be food-deprived, we compared the trained animals with similarly food-deprived untrained animals and normal controls. The results show that the excitability, short-term plasticity, and long term plasticity of the studied horizontal connections were unaffected by motor learning. However, stress-related effects produced by food deprivation and handling significantly enhanced the expression of long-term depression in these pathways. These results are compatible with the hypothesis that the acquisition of a complex motor skill produces bi-directional changes in synaptic strength that are distributed throughout the complex neural networks of motor cortex, which remains synaptically balanced during learning. The results are incompatible with the idea that learning causes large unidirectional changes in the population response of these neural networks, which may occur instead during certain behavioral states, such as stress. PMID- 15525805 TI - Glutamate neurotransmission is not required for, but may modulate, hypoxic sensitivity of pre-Botzinger complex in vivo. AB - Focal hypoxia in the pre-Botzinger complex (pre-BotC) in vivo elicits excitation of inspiratory motor output by modifying the patterning and timing of phrenic bursts. Hypoxia, however, has been reported to enhance glutamate release in some regions of the brain, including the medullary ventral respiratory column; thus the pre-BotC-mediated hypoxic respiratory excitation may result from, or be influenced by, hypoxia-induced activation of ionotropic glutamate [i.e., excitatory amino acid (EAA)] receptors. To test this possibility, the effects of focal pre-BotC hypoxia [induced by sodium cyanide (NaCN)] were examined before and after blockade of ionotropic EAA receptors [using kynurenic acid (KYN)] in this region in chloralose-anesthetized, vagotomized, mechanically ventilated cats. Before blockade of ionotropic EAA receptors, unilateral microinjection of NaCN (1 mM; 10-20 nl) into the pre-BotC produced either phasic or tonic excitation of phrenic nerve discharge. Unilateral microinjection of KYN (50-100 mM; 40 nl) decreased the amplitude and frequency of basal phrenic nerve discharge; however, subsequent microinjection of NaCN, but not DL-homocysteic acid (DLH, a glutamate analog), still produced excitation of phrenic motor output. Under these conditions, the NaCN-induced excitation included frequency modulation (FM) of phasic phrenic bursts, and in many cases, augmented and/or fractionated phrenic bursts. These findings show that the hypoxia-sensing function of the in vivo pre-BotC, which produces excitation of phrenic nerve discharge, is not dependent on activation of ionotropic glutamate receptors, but ionotropic glutamate receptor activation may modify the expression of the focal hypoxia-induced response. Thus these findings provide additional support to the concept of intrinsic hypoxic sensitivity of the pre-BotC. PMID- 15525806 TI - Hypercapnic exposure in congenital central hypoventilation syndrome reveals CNS respiratory control mechanisms. AB - Congenital central hypoventilation syndrome (CCHS) patients show impaired ventilatory responses and loss of breathlessness to hypercapnia, yet arouse from sleep to high CO2, suggesting intact chemoreceptor afferents. The syndrome provides a means to differentiate brain areas controlling aspects of breathing. We used functional magnetic resonance imaging to determine brain structures responding to inspired 5% CO2-95% O2 in 14 CCHS patients and 14 controls. Global signal changes induced by the challenge were removed on a voxel-by-voxel basis. A priori-defined volume-of-interest time trends (assessed with repeated measures ANOVA) and cluster analysis based on modeling each subject to a step function (individual model parameter estimates evaluated with t-test, corrected for multiple comparisons) revealed three large response clusters to hypercapnia distinguishing the two groups, extending from the 1) posterior thalamus through the medial midbrain to the dorsolateral pons, 2) right caudate nucleus, ventrolaterally through the putamen and ventral insula to the mid-hippocampus, and 3) deep cerebellar nuclei to the dorsolateral cerebellar cortex bilaterally. Smaller clusters and defined areas of group signal differences in the midline dorsal medulla, amygdala bilaterally, right dorsal-posterior temporal cortex, and left anterior insula also emerged. In most sites, early transient or sustained responses developed in controls, with little, or inverse change in CCHS subjects. Limbic and medullary structures regulating responses to hypercapnia differed from those previously shown to mediate loaded breathing ventilatory response processing. The findings show the significant roles of cerebellar and basal ganglia sites in responding to hypercapnia and the thalamic and midbrain participation in breathing control. PMID- 15525807 TI - Peaked encoding of relative luminance in macaque areas V1 and V2. AB - It is widely presumed that throughout the primate visual pathway neurons encode the relative luminance of objects (at a given light adaptation level) using two classes of monotonic function, one positively and the other negatively sloped. Based on computational considerations, we hypothesized that early visual cortex also contains neurons preferring intermediate relative luminance values. We tested this hypothesis by recording from single neurons in areas V1 and V2 of alert, fixating macaque monkeys during presentation of a large, spatially uniform patch oscillating slowly in luminance and surrounded by a static texture background. A substantial subset of neurons responsive to such low spatial frequency luminance stimuli in both areas exhibited prominent and statistically reliable response peaks to intermediate rather than minimal or maximal luminance values. When presented with static patches of different luminance but of the same spatial configuration, most neurons tested retained a preference for intermediate relative luminance. Control experiments using luminance modulation at multiple low temporal frequencies or reduced amplitude indicate that in the slow luminance oscillating paradigm, responses were more strongly modulated by the luminance level than the rate of luminance change. These results strongly support our hypothesis and reveal a striking cortical transformation of luminance-related information that may contribute to the perception of surface brightness and lightness. In addition, we tested many luminance-sensitive neurons with large chromatic patches oscillating slowly in luminance. Many cells, including the gray preferring neurons, exhibited strong color preferences, suggesting a role of luminance-sensitive cells in encoding information in three-dimensional color space. PMID- 15525808 TI - Intersegmental coordination of walking movements in stick insects. AB - Locomotion requires the coordination of movements across body segments, which in walking animals is expressed as gaits. We studied the underlying neural mechanisms of this coordination in a semi-intact walking preparation of the stick insect Carausius morosus. During walking of a single front leg on a treadmill, leg motoneuron (MN) activity tonically increased and became rhythmically modulated in the ipsilateral deafferented and deefferented mesothoracic (middle leg) ganglion. The pattern of modulation was correlated with the front leg cycle and specific for a given MN pool, although it was not consistent with functional leg movements for all MN pools. In an isolated preparation of a pair of ganglia, where one ganglion was made rhythmically active by application of pilocarpine, we found no evidence for coupling between segmental central pattern generators (CPGs) that could account for the modulation of MN activity observed in the semi intact walking preparation. However, a third preparation provided evidence that signals from the front leg's femoral chordotonal organ (fCO) influenced activity of ipsilateral MNs in the adjacent mesothoracic ganglion. These intersegmental signals could be partially responsible for the observed MN activity modulation during front leg walking. While afferent signals from a single walking front leg modulate the activity of MNs in the adjacent segment, additional afferent signals, local or from contralateral or posterior legs, might be necessary to produce the functional motor pattern observed in freely walking animals. PMID- 15525809 TI - Ipsilateral motor cortex activity during unimanual hand movements relates to task complexity. AB - Functional imaging studies have revealed recruitment of ipsilateral motor areas during the production of sequential unimanual finger movements. This phenomenon is more prominent in the left hemisphere during left-hand movements than in the right hemisphere during right-hand movements. Here we investigate whether this lateralization pattern is related specifically to the sequential structure of the unimanual action or generalizes to other complex movements. Using event-related fMRI, we measured activation changes in the motor cortex during three types of unimanual movements: repetitions of a sequence of movements with multiple fingers, repetitive "chords" composed of three simultaneous key presses, and simple repetitive tapping movements with a single finger. During sequence and chord movements, strong ipsilateral activation was observed and was especially pronounced in the left hemisphere during left-hand movements. This pattern was evident for both right-handed and, to a lesser degree, left-handed individuals. Ipsilateral activation was less pronounced in the tapping condition. The site of ipsilateral activation was shifted laterally, ventrally, and anteriorly with respect to that observed during contralateral movements and the time course of activation implied a role in the execution rather than planning of the movement. A control experiment revealed that strong ipsilateral activity in left motor cortex is specific to complex movements and does not depend on the number of required muscles. These findings indicate a prominent role of left hemisphere in the execution of complex movements independent of the sequential nature of the task. PMID- 15525811 TI - Activity of pyramidal tract neurons in the cat during postural corrections. AB - The dorsal side-up body orientation in quadrupeds is maintained by a postural control system. We investigated participation of the motor cortex in this system by recording activity of pyramidal tract neurons (PTNs) from limb representations of the motor cortex during postural corrections. The cat was standing on the platform periodically tilting in the frontal plane, and maintained equilibrium at different body configurations: with the head directed forward (symmetrically alternating loading of the left and right fore limbs), or with the head voluntary turned to the right or to the left (asymmetrical loading). We found that postural corrective responses to tilts included an increase of the contact forces and activity of limb extensors on the side moving down, and their decrease on the opposite side. The activity of PTNs was strongly modulated in relation to the tilt cycle. Phases of activity of individual PTNs were distributed over the cycle. Thus the cortical output mediated by PTNs appeared closely related to a highly automatic motor activity, the maintenance of the body posture. An asymmetrical loading of limbs, caused by head turns, resulted in the corresponding changes of motor responses to tilts. These voluntary postural modifications were also well reflected in the PTNs' activity. The activity of a part of PTNs correlated well with contact forces, in some others with the limb muscle activity; in still others no correlation with these variables was observed. This heterogeneity of the PTNs population suggests a different functional role of individual PTNs. PMID- 15525810 TI - Simultaneous contribution of two rod pathways to AII amacrine and cone bipolar cell light responses. AB - Rod signals traverse several synapses en route to cone bipolar cells. In one pathway, rods communicate directly with cones via gap junctions. In a second pathway, signals flow rods-rod bipolars-AII amacrines-cone bipolars. The relative contribution of each pathway to retinal function is not well understood. Here we have examined this question from the perspective of the AII amacrine. AIIs form bidirectional electrical synapses with on cone bipolars. Consequently, as on cone bipolars are activated by outer plexiform inputs, they too should contribute to the AII response. Rod bipolar inputs to AIIs were blocked by AMPA receptor antagonists, revealing a smaller, non-AMPA component of the light response. This small residual response did not reverse between -70 and +70 mV and was blocked by carbenoxolone, suggesting that the current arose in on cone bipolars and was transmitted to AIIs via gap junctions. The residual component was evident for stimuli 2 log units below cone threshold and was prolonged for bright stimuli, demonstrating that it was rod driven. Because the rod bipolar-AII pathway was blocked, the rod-driven residual current likely was generated via the rod-cone pathway activation of on cone bipolars. Thus for a large range of intensities, rod signals reach the inner retina by both rod bipolar-AII and rod-cone coupling pathways. PMID- 15525812 TI - Canal-otolith interactions after off-vertical axis rotations. II. Spatiotemporal properties of roll and pitch postrotatory vestibuloocular reflexes. AB - We have examined the spatiotemporal characteristics of postrotatory eye velocity after roll and pitch off-vertical axis rotations (OVAR). Three rhesus monkeys were placed in one of 3 orientations on a 3-dimensional (3D) turntable: upright (90 degrees roll or pitch OVAR), 45 degrees nose-up (45 degrees roll OVAR), and 45 degrees left ear-down (45 degrees pitch OVAR). Subjects were then rotated at +/-60 degrees /s around the naso-occipital or interaural axis and stopped after 10 turns, in one of 7 final head orientations, each separated by 30 degrees . We found that postrotatory eye velocity showed horizontal-vertical components after roll OVAR and horizontal-torsional components after pitch OVAR that varied systematically as a function of final head orientation. The quantitative analysis suggests that, in contrast to the analogous yaw OVAR paradigm, a system of up to 3 real, gravity-dependent eigenvectors and eigenvalues determines the spatiotemporal characteristics of the residual eye velocities after roll and pitch OVAR. One of these eigenvectors closely aligned with gravity, whereas the other 2 determined the orientation of the earth horizontal plane. We propose that the spatial characteristics of eye velocity after roll and pitch OVAR follow the physical constraints of stationary orientation in a gravitational field and reflect the brain's best estimate of head-in-space orientation within an internal representation of 3D space. PMID- 15525814 TI - Percoll gradient-centrifuged capacitated mouse sperm have increased fertilizing ability and higher contents of sulfogalactosylglycerolipid and docosahexaenoic acid-containing phosphatidylcholine compared to washed capacitated mouse sperm. AB - Although Percoll gradient centrifugation has been used routinely to prepare motile human sperm, its use in preparing motile mouse sperm has been limited. Here, we showed that Percoll gradient-centrifuged (PGC) capacitated mouse sperm had markedly higher fertilizing ability (sperm-zona pellucida [ZP] binding and in vitro fertilization) than washed capacitated mouse sperm. We also showed that the lipid profiles of PGC capacitated sperm and washed capacitated sperm differed significantly. The PGC sperm had much lower contents of cholesterol and phospholipids. This resulted in relative enrichment of male germ cell-specific sulfogalactosylglycerolipid (SGG), a ZP-binding ligand, in PGC capacitated sperm, and this would explain, in part, their increased ZP-binding ability compared with that of washed capacitated sperm. Analyses of phospholipid fatty acyl chains revealed that PGC capacitated sperm were enriched in phosphatidylcholine (PC) molecular species containing highly unsaturated fatty acids (HUFAs), with docosahexaenoic acid (DHA; C22: 6n-3) being the predominant HUFA (42% of total hydrocarbon chains of PC). In contrast, the level of PC-HUFAs comprising arachidonic acid (20:4n-6), docosapentaenoic acid (C22:5n-6), and DHA in washed capacitated sperm was only 27%. Having the highest unsaturation degree among all HUFAs in PC, DHA would enhance membrane fluidity to the uppermost. Therefore, membranes of PGC capacitated sperm would undergo fertilization-related fusion events at higher rates than washed capacitated sperm. These results suggested that PGC mouse sperm should be used in fertilization experiments and that SGG and DHA should be considered to be important biomarkers for sperm fertilizing ability. PMID- 15525813 TI - Estrogen and inflammation increase the excitability of rat temporomandibular joint afferent neurons. AB - Several painful conditions, including temporomandibular disorders (TMD), are more prevalent and more severe in women than in men. Although the physiological basis for this sex difference remains to be determined, it is likely that estrogen is an underlying factor. The present study was performed to test the hypotheses that estrogen increases the excitability of rat temporomandibular joint (TMJ) afferents and exacerbates the inflammation-induced sensitization of these sensory neurons. Retrogradely labeled TMJ neurons from ovariectomized rats and ovariectomized rats receiving chronic estrogen replacement were studied using whole cell patch-clamp techniques three days after injecting the TMJ with either saline or Complete Freund's Adjuvant to induce inflammation. Excitability was assessed with depolarizing current injection to determine action potential threshold, rheobase, and the response to suprathreshold stimuli. Spontaneous activity was also assessed. Both inflammation and estrogen increased the excitability of TMJ neurons as reflected by decreases in action potential threshold and rheobase and increases in the incidence of spontaneous activity. The effects were additive with neurons from rats receiving both estrogen and inflammation being the most excitable. The increases in excitability were associated with changes in passive properties and action potential waveform, suggesting that estrogen and inflammation affect the expression and/or properties of ion channels in TMJ neurons. Importantly, the influence of estrogen on both baseline and inflammation-induced changes in TMJ neuronal excitability may help explain the profound sex difference observed in TMD as well as suggest a novel target for the treatment of this pain condition. PMID- 15525815 TI - Reproductive hormones and follicular growth during development of one or multiple dominant follicles in cattle. AB - The mechanisms regulating ovulation rate under natural conditions are not yet defined, particularly for monovular species. In the present study, we evaluated ovarian structures (every 12 h by ultrasonography) and circulating hormones (every 6 h) to determine the differences between cows that developed one (single dominant; n = 16), two (double dominant; n = 8), or three (triple dominant; n = 3) dominant follicles. The four largest follicles were tracked retrospectively, and the data were normalized to the time of expected follicular deviation (F1 >/= 8.5 mm; hour 0). Follicular dynamics from emergence to deviation were similar, whereas after deviation, expected subordinate follicles continued to grow at a rate similar to the dominant follicle. Triple dominants had greater FSH than double dominants (hour -24 to hour -12) and single dominants (hour -42 to hour 6), and double dominants had greater FSH than single dominants (hour -24 to hour 12). Increased circulating estradiol but lower inhibin were observed in cows that developed multiple follicles. In addition, double dominants had greater LH than single dominants (hour -42 to hour -24 and hour -6 to hour 0) and lower progesterone than single dominants (hour -12 and hour -6). Luteal volume was similar between groups, but milk production was greater for codominant than for single-dominant cows. Thus, selection of multiple dominant follicles during high milk production is related to a transient increase in circulating FSH and LH during the 24 h before follicular selection, producing continued postdeviation growth of follicles that ordinarily would have regressed. Increased FSH and LH probably result from decreased circulating inhibin and progesterone in cows that develop codominant follicles. PMID- 15525816 TI - Possibility of long-term preservation of freeze-dried mouse spermatozoa. AB - Freeze-dried mouse spermatozoa are capable of participating in normal embryonic development after injection into oocytes. When the freeze-dried spermatozoa are used as a method for storage of genetic materials, however, it is essential to assure the relevance of long-term preservation over several decades or centuries. Thus, we applied the theory of accelerated degradation kinetics to freeze-dried mouse spermatozoa. Thermal denaturation kinetics were determined based on Arrhenius plots derived from transition-state theory analysis at three elevated temperatures: 30, 40, and 50 degrees C. Accelerated degradation kinetics were calculated by extrapolation of Arrhenius plots. This theory also is being applied to the long-term stability of drugs. The estimated rate of development to the blastocyst stage at 3 and 6 mo and at 1, 10, and 100 yr of sperm storage at 4 degrees C were 21.60%, 7.91%, 1.00%, 0%, and 0%, respectively. At -80 degrees C, estimated development rates to the blastocyst stage that would be expected after 100 yr of storage did not decline significantly. In addition, after 3 or 6 mo of storage at 4 or -80 degrees C, preimplantation development of the embryos derived from intracytoplasmic sperm injection (ICSI) was examined. The actual developmental rates to the blastocyst stage from ICSI by freeze-dried sperm stored for 3 mo at 4 and -80 degrees C were 21% and 62%, respectively, and the rates for such sperm stored for 6 mo were 13% and 59%, respectively. These results indicate that the determination of accelerated degradation kinetics can be applied to the preservation of freeze-dried mouse spermatozoa. Furthermore, for long-term preservation, freeze-dried mouse spermatozoa appear to require being kept at lower than -80 degrees C. PMID- 15525817 TI - Effect of treating induced mitochondrial damage on embryonic development and epigenesis. AB - Germinal vesicle transplantation (GVT) has been proposed as a possible treatment to correct age-related oocyte aneuploidy caused by dysfunctional ooplasm. How healthy ooplasm regulates normal meiosis and subsequent development has yet to be elucidated, but impaired mitochondrial metabolism may be attributable to incomplete segregation of the oocyte chromosomes. In the present study, after ooplasmic mitochondrial damage by photoirradiating chloromethyl-X-rosamine, examination of the oocyte nuclei's ability to survive after transfer into healthy ooplasts was performed. To assess their fertilizability and potential for development, GVT oocytes were fertilized by intracytoplasmic sperm injection (ICSI) and transferred to foster mice. Condition of the offspring at birth was assessed, and epigenetic analysis was performed. Photosensitization consistently inhibited oocyte maturation. However, after GVT of photosensitized nuclei into healthy ooplasts, 67.2% were reconstituted, and 76.2% of these matured normally, with an overall rate of 51.2%, much higher than that (6.0%) in the mitochondrially injured oocytes. After ICSI, 65.8% (52/79) of GVT oocytes were fertilized normally, and 21.1% (11/52) eventually reached the blastocyst stage. The transfer of 132 two-cell GVT embryos into the oviducts of pseudopregnant females resulted in 17 apparently healthy live offspring. For some key developmental genes, a high level of expression was identified in the GVT and "rescue"-derived fetal adnexa. Thus, one can induce in oocyte mitochondria a photosensitization-based type of damage, which consistently inhibits GV breakdown, meiotic spindle formation, chromosomal segregation, and polar body extrusion. Germinal vesicle transplanted and rescued oocytes were able to undergo maturation, fertilization, and embryonic cleavage and, ultimately, to develop to term. This approach may provide a model with which to study the age-related ooplasmic dysfunction seen in human oocytes. PMID- 15525818 TI - Increased expression of protein C-mannosylation in the aortic vessels of diabetic Zucker rats. AB - C-Mannosylation is a novel type of glycosylation in proteins. There are several examples of proteins in which the specific motif Trp-X-X-Trp is mannosylated at the first Trp to produce C-mannosylated Trp (CMW). Although C-mannosylation modifies Trp-X-X-Trp, predicted to be a functional motif of various integral proteins such as cytokine receptors, the physiological or pathological relevance of C-mannosylation in the cell is still not known. In this study, to characterize C-mannosylation in biological samples, we generated specific polyclonal antibodies against CMW by using a chemically synthesized CMW as an antigen. Using the antibody, we investigated the effect of hyperglycemic conditions on protein C mannosylation in cultured cells and diabetic Zucker fatty rats. We found that protein C-mannosylation was increased in macrophage-like RAW264.7 cells under hyperglycemic conditions compared to low-glucose conditions. Furthermore, C mannosylation was increased in the aortic vessel wall of Zucker fatty rats. Thrombospondin-1 was identified as a protein modified with C-mannosylation, and its expression was also increased in the aortic tissues of Zucker fatty rats. These results indicate that C-mannosylation is increased in specific tissues or cell types under hyperglycemic conditions, suggesting a pathological role for the increased C-mannosylation in the development of diabetic complications. PMID- 15525819 TI - Suppressors of alpha(1,3)fucosylation identified by expression cloning in the LEC11B gain-of-function CHO mutant. AB - Factors that regulate alpha(1,3)fucosyltransferase activity are important to identify because FUT genes are up-regulated during inflammation, cancer progression, and tumor metastasis. FUT gene activation increases the expression of cell surface oncofetal antigens such as Lewis X, sialyl-Le X and VIM-2. The LEC11B gain-of-function glycosylation mutant displays these antigens and binds E selectin because it expresses the Fut6B gene that is shown here to lie immediately downstream of the Fut6A gene. A retroviral strategy for expression cloning factors that suppress alpha(1,3)fucosylation in LEC11B cells was developed, and several cDNAs that reverted the LEC11B glycosylation phenotype were isolated. cDNAs that arose most frequently and independently encoded SLC35C2, a putative GDP-fucose transporter (also termed CGI-15 or Ovcov1); Cd63, a tetraspanin membrane protein; and Hdac5, a histone deacetylase. When transfected into LEC11B cells the SLC35C2 cDNA reduced Le X expression with no concomitant suppression of Fut6B gene transcripts. Transfection of the Cd63 cDNA induced low levels of ricin resistance and also did not suppress Fut6B gene transcripts in LEC11B. However, the Hdac5 cDNA induced ricin resistance, reduced fucosylated antigen expression, and essentially eliminated Fut6B gene transcripts. The Hdac5 cDNA isolated by expression cloning encoded the C-terminal region of hamster Hdac5. Overexpression of this partial Hdac5 cDNA or a full length Hdac5 cDNA, suppressed Fut6B gene transcripts specifically. Thus the expression cloning strategy identified Hdac5 as a trans-acting repressor of the Chinese hamster ovary Fut6B gene and Cd63 and SLC35C2 as novel factors that suppress alpha(1,3)fucosylation by mechanisms unrelated to effects on Fut gene expression. PMID- 15525820 TI - Markers of cell cycle-mediated drug resistance and prognosis of patients receiving preoperative combined modality therapy for rectal cancer. PMID- 15525821 TI - Age and cancer surgery: judicious selection or discrimination? PMID- 15525822 TI - Molecular staging of melanoma. PMID- 15525823 TI - Expression of p27 in residual rectal cancer after preoperative chemoradiation predicts long-term outcome. AB - BACKGROUND: Compared with surgery alone, preoperative radiotherapy and 5 fluorouracil-based chemotherapy (combined-modality therapy; CMT) improves outcomes in patients with locally advanced rectal cancer. Although numerous studies have focused on identifying molecular markers of prognosis in the primary rectal cancer before CMT, our aim was to identify markers of prognosis in residual rectal cancer after preoperative CMT. METHODS: Sixty-seven patients with locally advanced (T3-4 and/or N1) rectal cancer were treated with preoperative radiotherapy (median, 5040 cGy) with or without 5-fluorouracil-based chemotherapy. Residual tumor in the resected specimen, available for 52 patients, was analyzed for tumor-node-metastasis stage, lymphovascular and/or perineural invasion, and immunohistochemical expression of p27, p21, p53, Ki-67, retinoblastoma gene, cyclin D1, and bcl-2. Recurrence-free survival (RFS) was determined by the Kaplan-Meier method and compared by the log-rank test. RESULTS: With a median follow-up of 69 months, the overall 5-year RFS was 74%. RFS was significantly worse for patients with positive p27 expression (P = .005), T3-4 tumors (P = .02), and positive lymph nodes (P = .04) in the irradiated specimen. On multivariate analysis, positive p27 expression remained an independent negative prognostic factor for RFS (P = .04). None of the other proteins was significantly associated with RFS. CONCLUSIONS: Our results indicate that positive p27 expression in rectal cancer after preoperative chemoradiation is an independent negative predictor of RFS. Expression of p27 in the residual rectal cancer may therefore identify patients with disease likely to be refractory to standard therapy and for whom investigational approaches should be strongly considered. PMID- 15525824 TI - Cancer-directed surgery for localized disease: decreased use in the elderly. AB - BACKGROUND: Previous studies report underuse of radiation and chemotherapy in the elderly, yet few have examined the rates of use (or underuse) of surgery. Using national data, we examined rates of surgical resection for patients with local stage cancers. METHODS: By using the Surveillance, Epidemiology, and End RESULTS database (1988-1997), patients (> or =40 years) were identified with localized adenocarcinoma of the breast, esophagus, stomach, pancreas, colon, or rectum; non small-cell lung carcinoma; and sarcoma (n = 200,360). Rates of cancer-directed surgery (CDS) were compared across age groups (at 5-year intervals). Multivariate regression was used to identify predictors of receipt of CDS in each tumor group. RESULTS: Rates of CDS declined steadily with increasing age for all nine localized tumors. Most striking were the low rates of CDS for patients >70 years with esophagus, stomach, pancreas, and lung cancers (range, 0%-83%). However, CDS rates were >90% for breast and colon and >84% for rectal cancer in all age groups. Multivariate regression found lower odds of CDS for elderly patients for all cancers except colon. For example, age significantly decreased the odds of receiving CDS beginning at 60 years for lung cancer (odds ratio [OR], .550; P = .03), at 70 years for liver cancer (OR, .109; P = .003), and at 80 years for pancreatic cancer (OR, .120; P < .05). CONCLUSIONS: Although CDS for localized disease is being performed regularly in the elderly for some cancers (e.g. breast, colon, and rectum), this analysis shows that elderly patients are not receiving surgery for many potentially curable cancers. Whether these rates are appropriate or too low requires further evaluation. This is particularly essential because our population is aging. PMID- 15525825 TI - Institutional commitment to rectal cancer screening results in earlier-stage cancers on diagnosis. AB - BACKGROUND: The Veterans Administration hospitals underwent an institutional directive in 1998 to meet a colorectal cancer screening (CRCS) standard. This intervention should result in an increase in the hospital's screening rate and percentage of early-stage rectal cancers diagnosed. METHODS: A retrospective review, from 1991 to 2002, of our institution's pathology and cancer registry databases for rectal cancers. CRCS data were obtained from the Veterans Administration Prevention Disease Index. Cancer stage at diagnosis was compared before and after the directive and was compared with the National Cancer Data Base and the Surveillance, Epidemiology, and End Results data. RESULTS: The rate of CRCS was 55% in 1998 and increased to 75% in 2003. During the 11 years studied, a total of 147 rectal cancers were diagnosed. After the Veterans Administration directive, there was a significant increase in stage 0 cancers (P < .02) and an overall migration to earlier-stage cancers. Our Veterans Administration hospital had a significantly greater percentage of stage 0 cancers both before (P < .007) and after the directive (P < .00) and had fewer stage 3 cancers after the directive (P < .03) compared with National Cancer Data Base data. Compared with Surveillance, Epidemiology, and End Results data, the Palo Alto Veterans Affairs Health Care System had more local disease (P < .03) and less regional disease (P < .006). CONCLUSIONS: These data suggest that a monitored institutional directive may significantly increase early detection of rectal cancers. This should result in a greater survival rate and lower local recurrence rate, because survival is predicated on stage at presentation. This may serve as a model for other health-care systems. PMID- 15525826 TI - Microsatellite instability as a prognostic factor in resected colorectal cancer liver metastases. AB - BACKGROUND: Two distinct genetic mutational pathways characterized by either chromosomal instability or high-frequency microsatellite instability (MSI-H) are currently recognized in the pathogenesis of colorectal cancer (CRC). Recently, it has been shown that patients with primary CRC that displays MSI-H have a significant, stage-independent, multivariate survival advantage. Untreated CRC hepatic metastases are incurable and are associated with a median survival of 4 to 12 months. Conversely, surgical resection in selected patients results in a 20% to 50% cure rate. The aim of this study was to investigate the prognostic importance of MSI-H in patients undergoing resection of hepatic CRC metastases. METHODS: DNA was extracted from paraffin-embedded, resected metastatic CRC liver lesions and corresponding normal liver parenchyma from 190 patients. MSI-H status was determined by polymerase chain reaction-based evaluation of the noncoding mononucleotide repeats BAT-25 and BAT-26. RESULTS: MSI was detected in tumors from 5 (2.7%) of the 190 CRC patients. All MSI-H tumors were in patients with node-positive CRC primary tumors. The median survival after hepatic resection of MSI-H and non-MSI-H tumors was 67 and 61 months, respectively (P = .9). CONCLUSIONS: These data suggest that MSI-H is not a common feature in resected CRC liver metastases and do not suggest a role for MSI in stratifying good versus poor prognosis in these patients. PMID- 15525827 TI - Detection of melanoma cells in sentinel lymph nodes by reverse transcriptase polymerase chain reaction: prognostic significance. AB - BACKGROUND: Recently reverse transcriptase-polymerase chain reaction (RT-PCR) has been proposed as a new sensitive method for the detection of submicroscopic melanoma nodal metastases. Sentinel lymph node (SLN) status is considered the most important prognostic factor for melanoma patients. Thus, in recent years, melanoma research has been focused on identifying new molecular markers of micrometastases. METHODS: In this study, 129 SLNs were collected and analyzed by RT-PCR for tyrosinase and melanoma inhibitory activity (MIA) messenger RNA (mRNA) expression. RESULTS from PCR analysis were then compared with those obtained by hematoxylin and eosin and immunohistochemistry and related to progression of disease. RESULTS: MIA gene expression was positive by RT-PCR in 27% of the tyrosinase-positive SLNs. When the correlation between tyrosinase and/or MIA mRNA expression and disease-free survival was evaluated by the Kaplan-Meier exact test, there was a statistically significant correlation between simultaneous tyrosinase and MIA gene expression in SLNs and progression of disease. CONCLUSIONS: RT-PCR analysis for both MIA and tyrosinase mRNA may identify a subset of melanoma patients with a worse prognosis whom the routine methods, such as histology and immunohistochemistry, fail to identify because of the poor sensitivity of these methods. PMID- 15525828 TI - Detection of melanoma cells in the lymphatic drainage after lymph node dissection in melanoma patients by using two-marker reverse transcriptase-polymerase chain reaction assay. AB - BACKGROUND: The aim of this study was to evaluate the role of melanoma gene expression as a marker of the presence of melanoma cells in lymphatic drainage routinely collected after lymphadenectomy and to correlate reverse transcriptase polymerase chain reaction (RT-PCR) assay results with recurrence, survival, and prognostic factors. METHODS: We collected 24-hour postoperative lymphatic drainage samples (between days 2 and 4) from 93 patients with stage III melanoma who underwent radical lymphadenectomy between May 2002 and November 2003. We used RT-PCR assays with primers specific for the tyrosinase and MART-1 (Melan-A) genes. The samples were considered positive if at least one marker was expressed. Median follow-up time was 12.8 months. RESULTS: In 18 (19.4%) of 93 patients, the RT-PCR assay results were positive: in 8 of 18 for tyrosinase only, in 7 of 18 for MART-1 only, and in 3 of 18 for both markers. We observed a significantly higher recurrence rate in patients with positive RT-PCR results (15 of 18; 83%) than negative results (26 of 75; 35%; P = .0001). Positive results of RT-PCR correlated with the number of involved lymph nodes (P = .0001) and extracapsular extension of nodal metastases (P = .03). We observed significant differences in overall and disease-free survival for RT-PCR-positive and -negative patients in univariate and multivariate analyses. CONCLUSIONS: We observed positive RT-PCR assay results for melanoma cells in the lymphatic drainages of approximately 20% of patients after lymphadenectomy. This correlated significantly with early recurrence and shorter survival. These results may suggest that the RT-PCR assay could be useful for routinely analyzing postoperatively collected lymphatic drainage in stage III melanoma patients and for predicting disease progression. PMID- 15525829 TI - Osteosarcomas after the age of 50: a clinicopathologic study of 64 cases--an experience in northern Japan. AB - BACKGROUND: The clinicopathologic profile and prognostic factors of osteosarcomas after the age of 50 years have been obscure. METHODS: Clinicopathologic features were analyzed in 645 patients with osteosarcoma who were registered at the Tohoku Musculoskeletal Tumor Society and National Cancer Center in Tokyo between 1972 and 2002. RESULTS: Sixty-four patients (9.9%; 34 men and 30 women) were more than 50 years old. The most common location was the distal femur (n = 13), followed by the pelvis (n = 10), proximal femur (n = 9), and proximal fibula (n = 6). Seven (11%) patients had lung metastasis at initial presentation. On radiographs, an osteolytic appearance without periosteal reactions was a common and characteristic feature. Forty-eight tumors (75%) were classified as conventional osteosarcomas. Fourteen cases (22%) were secondary; postradiation osteosarcoma was most common in these patients, but there was no Paget's sarcoma. At the initial presentation, misdiagnoses without biopsy were made in 15 (23%) of the 64 cases. Preoperative chemotherapy was given to 22 patients, but the effect was poor in 18 cases (82%). Fifty-four patients underwent surgery, whereas the other 10 patients were treated without surgery because of systemic or inoperable local conditions. The overall survival rate at 5 years was 55.5%. Multivariate analysis showed initial pulmonary metastasis, axial tumor location, and larger tumors as significant prognostic factors. CONCLUSIONS: In northern Japan, most patients with osteosarcoma after the age of 50 had primary osteosarcoma. Careful radiological examination and biopsy are mandatory for correct diagnosis. Current systemic chemotherapy is not effective for this age group. Alternative treatment strategies should be considered. PMID- 15525830 TI - Intraoperative examination of sentinel nodes in breast cancer: is the glass half full or half empty? AB - BACKGROUND: Intraoperative identification of positive sentinel lymph nodes in patients with breast cancer may avoid a return to the operating room. METHODS: In a group of 402 consecutive patients with primary breast cancer who underwent sentinel lymph node biopsy, an intraoperative examination (IE) was obtained in 236 cases either by frozen section (FS; n = 68) or by touch preparation cytology (TP; n = 168). RESULTS: IE had an accuracy of 89% (209 of 236), but it identified only 52 of 77 positive cases (sensitivity, 68%). There were 25 false-negative cases (13.7%), of which 7 were macrometastases and 18 by micrometastases (P < .001). Six macrometastases were missed by TP and one by FS (P = .9). There were two false-positive cases (3.7%). Overall, 48 (20%) of 236 patients avoided a delayed return to the operating room for a completion lymphadenectomy because of IE findings. This occurred in 10% of patients with tumors <1 cm in diameter, in 20% of those with tumors between 1 and 2 cm, and in 34% of those with tumors >2 cm in diameter (P = .05). The cost savings for the Italian Health System amounted to 198,040 (US$223,794) in these patients. CONCLUSIONS: IE has acceptable sensitivity for lymph node macrometastases, but it is a weak tool for diagnosing micrometastases. FS and TP are roughly equivalent. IE allows management changes, because approximately 20% of all patients are expected to undergo synchronous axillary dissection, and it is particularly helpful in T2 patients. This may allow substantial cost savings for the health-care system. PMID- 15525831 TI - Risk factors for recurrence and death after primary surgical treatment of malignant phyllodes tumors. AB - BACKGROUND: Malignant phyllodes tumor (MPT) is a rare but aggressive breast malignancy. The aim of this study was to evaluate parameters that influence outcome in patients with MPT. METHODS: Fifty women were diagnosed with MPT of the breast and treated between August 1971 and July 2000. All medical records were reviewed retrospectively. The Cox regression model was used for multivariate analysis. RESULTS: Tumors were classified as borderline (6%), low grade (32%), or high grade (62%). The median patient age was 46 years (range, 14-77 years). The median tumor diameter was 3.5 cm (range, 1.5-18 cm). Twenty-two patients had wide local excision (WLE), and 28 patients had mastectomy. The median follow-up was 91 months (range, 12-360 months). Local recurrence (LR) occurred in 16 patients (32%) an average of 26 months after surgery (median, 17 months; range, 3-72 months). Distant metastasis occurred in 13 patients (26%) at an average of 53.4 months (median, 36 months; range, 4-177 months). Sixteen (32%) patients have died of their disease. LR was significantly increased with stromal overgrowth (P < .0001), large tumor size (P = .0177), and surgical margins <1 cm (P = .0120), but not with WLE (P = .5099). Stromal overgrowth was the only independent variable predictive of systemic metastasis (P < .0001) and patient survival (P < .0001). CONCLUSIONS: Stromal overgrowth in MPT carries a grave prognosis. Close surgical margins and large tumor size, but not type of operation, significantly increased LR. Either WLE with adequate margins or mastectomy is an appropriate treatment for patients with MPT. PMID- 15525834 TI - Relay man. Molecule transmits estrogen's bone-saving signal through T cells. PMID- 15525832 TI - Treatment-related upper limb morbidity 1 year after sentinel lymph node biopsy or axillary lymph node dissection for stage I or II breast cancer. AB - BACKGROUND: In a prospective study, upper limb morbidity and perceived disability/activities of daily life (ADLs) were assessed before and 1 year after sentinel lymph node biopsy (SLNB) or axillary lymph node dissection (ALND). METHODS: A total of 204 patients with stage I/II breast cancer (mean age, 55.6 years; SD, 11.6 years) entered the study, and 189 patients (93%) could be evaluated after 1 year. Fifty-eight patients (31%) underwent only SLNB, and 131 (69%) underwent ALND. Assessments performed before surgery (t(0)) and 1 year after surgery (t(1)), included pain, shoulder range of motion, muscle strength, upper arm/forearm circumference, and perceived shoulder disability/ADL. RESULTS: Considerable treatment-related upper limb morbidity was observed. Significant (P < .05) changes between t(0) and t(1) were found in all assessments except strength of elbow flexors. Patients in the ALND group showed significantly more changes in the range of motion in forward flexion, abduction, and abduction/external rotation; grip strength and strength of shoulder abductors; circumference of upper arm and forearm; and perceived shoulder disability in ADLs compared with the SLNB group. Multivariate linear regression analysis showed that ALND could predict a decrease of range of motion in forward flexion, abduction, strength of shoulder abductors, grip strength, and shoulder-related ADLs and an increase in the circumference of the upper arm. Radiation of the axilla (19 patients) predicts an additional decrease in shoulder range of motion. CONCLUSIONS: One year after treatment of breast cancer, there is significantly less upper limb morbidity after SLNB compared with ALND. ALND is a predictor for upper limb morbidity. PMID- 15525835 TI - Gut instinct. Intestinal microbes promote fat buildup in mice. PMID- 15525836 TI - The search for biomarkers of aging: next stop INK4a/ARF locus. AB - Although several biomarkers of aging have been described in the literature, it is only recently that gerontologists have started to search for molecular biomarkers of aging. A gene or a set of genes that are expressed in a wide range of tissues and exhibit an age-dependent, easily quantifiable increase in their expression represent a possible molecular biomarker of aging. Because the physiology of an organism is profoundly affected by the pattern of gene expression, it is hoped that molecular biomarkers of aging will more accurately predict the physiological age of an organism than the chronological age. A recent report from Sharpless's laboratory examines the possibility that the tumor suppressors p16 and ARF (encoded by the INK4a/ARF locus) represent molecular biomarkers of aging in rodent models. PMID- 15525837 TI - Evidence for increased risk of secondary graft failure after in vivo depletion of suicide gene-modified T lymphocytes transplanted in conjunction with CD34+ enriched blood stem cells. PMID- 15525838 TI - Identification of immunodominant alloreactive T-cell epitopes on the Jka red blood cell protein inducing either Th1 or Th2 cytokine expression. PMID- 15525839 TI - Chondrogenic differentiation of mesenchymal stem cells is inhibited after magnetic labeling with ferumoxides. PMID- 15525840 TI - Important publication missing key information. PMID- 15525841 TI - Fatal case of protothecosis in a hematopoietic stem cell transplant recipient after infliximab treatment for graft-versus-host disease. PMID- 15525842 TI - Diamond-Blackfan anemia responding to valproic acid. PMID- 15525844 TI - Cardiac sarcoidosis: a retrospective study of 41 cases. AB - This retrospective study concerned 18 female and 23 male patients with cardiac sarcoidosis (CS). The average age at CS diagnosis was 38 years. CS was observed in white (73% of cases) and in black or Caribbean patients (27% of cases). All patients had extracardiac histologic proof of sarcoid tissue. In 63% of cases, the CS arose during the follow-up of systemic sarcoidosis. Systemic sarcoidosis was not specific except for a high frequency of neurosarcoidosis. Revealing cardiac signs were clinical in 63% of cases and electrical in 22%. In most patients these signs were associated with an abnormal echocardiography (77%) and/or a defect on thallium-201 or sestamibi imaging (75%). Thirty-nine patients received steroid therapy (initial dose mostly equal to 1 mg/kg per day), associated in 13 cases with another immunosuppressive treatment. In 26% of cases the immunosuppressive treatment was associated with a specific cardiac treatment. In the long-term follow-up (average follow-up, 58 mo), 87% of the cases showed an improvement, and 54% were cured from a clinical and laboratory point of view (electrocardiogram, 24-hour monitoring, echocardiography, radionuclide imaging). There was no sudden death. Two patients worsened, which can be explained in 1 case by very late treatment and in the other case by lack of treatment, except for a pacemaker. Our experience leads us to treat CS with corticosteroids as soon as possible and to use another immunosuppressive treatment where there is an insufficient therapeutic response or where there are contraindications to corticosteroids. PMID- 15525845 TI - Aortic aneurysm and dissection in patients with biopsy-proven giant cell arteritis from northwestern Spain: a population-based study. AB - Most classical manifestations of giant cell arteritis (GCA) are the result of occlusive vascular involvement. However, unlike ischemic manifestations, aortic aneurysmal disease in patients with GCA has been less well described. We assessed the incidence and predictors of aortic aneurysm and dissection in patients with biopsy-proven GCA from the Lugo region of northwestern Spain and compared the results with those in a 2003 report from Olmsted County, MN. We performed a retrospective study of biopsy-proven GCA patients diagnosed from 1981 to 2001 at the single hospital for a well-defined population of almost 250,000 people. Twenty (9.5%) of the 210 biopsy-proven GCA patients diagnosed during the study period developed aortic aneurysmal disease. Sixteen of the 20 patients had thoracic aneurysms and 6 had abdominal aneurysms. The incidence of aortic aneurysm and/or dissection in Lugo (18.9 per 1000 person years at risk) was similar to that reported in Olmsted County (18.7 per 1000 person years at risk). Hypertension (hazard ratio: 4.73) and polymyalgia rheumatica with a marked acute inflammatory response at the time of diagnosis of GCA (hazard ratio: 3.71) were the best predictors of aortic aneurysmal disease. Our present observations suggest that a severe inflammatory response associated with hypertension at the time of diagnosis of GCA may promote the development of aortic aneurysmal disease. GCA patients having these features should be monitored for the existence of aortic aneurysm and dissection. PMID- 15525846 TI - Influence of traditional risk factors of atherosclerosis in the development of severe ischemic complications in giant cell arteritis. AB - Because the prognosis of giant cell arteritis (GCA) is related to the development of ischemic complications, we sought to assess the possible influence of traditional risk factors of atherosclerosis in the development of severe ischemic complications of GCA. We conducted a retrospective study of patients with biopsy proven GCA diagnosed from 1981 to 2001 at the single hospital for a well-defined population of almost 250,000 people. Patients were considered to have severe ischemic manifestations if they suffered visual manifestations, cerebrovascular accidents, jaw claudication, or signs of occlusive changes in large arteries of the extremities. Patients were assessed for the presence of hypercholesterolemia, hypertension, diabetes mellitus, and heavy smoking at the time of GCA diagnosis. The presence of traditional risk factors of atherosclerosis at the time of GCA diagnosis in this series of 210 patients increased significantly the risk of developing at least 1 of the severe ischemic complications (odds ratio [OR], 1.79; 95% confidence intervals [CI], 1.03-3.11; p = 0.04). Patients with traditional atherosclerosis risk factors had fever less commonly than the rest of GCA patients (5.2% vs. 16.0%; p = 0.01). GCA patients with hypertension exhibited a significantly increased risk of developing severe ischemic complications (OR, 1.80; 95% CI, 1.00-3.25; p = 0.05). The current study suggests that the presence of atherosclerosis risk factors at the time of diagnosis of GCA may influence the development of severe ischemic manifestations of the disease. PMID- 15525847 TI - Late-onset systemic lupus erythematosus: a personal series of 47 patients and pooled analysis of 714 cases in the literature. AB - Systemic lupus erythematosus (SLE) is uncommon after the age of 50 years, and studies of elderly patients with SLE are scarce. We conducted the current study to analyze characteristics and outcome of patients with late-onset SLE in a French tertiary referral center, and to compare them with those of younger patients with SLE. From 1980 to 2000, 47 patients were identified as having late onset SLE, defined as SLE diagnosed at or over the age of 50 years. These patients were compared with a group of 114 randomly selected patients aged younger than 50 years at SLE diagnosis. We compared clinical characteristics, laboratory data, therapy, and course. The female to male ratio was smaller in the late-onset SLE group (p = 0.0012). Some manifestations occurred less frequently in late-onset SLE: arthritis (p = 0.009), malar rash (p = 0.013), and nephropathy (p = 0.009). High-dose corticosteroids (p = 0.0016) and immunosuppressive drugs (p = 0.006) were less commonly used in the elderly. Deaths occurred more frequently in late-onset SLE (p = 0.019), with a 10-year survival rate of 71% versus 95% in early-onset SLE (p < 0.01). In patients with late-onset SLE, causes of death were usually unrelated to SLE. Analysis of pooled data from the literature, based on 714 old and 4700 young SLE patients, confirmed that late onset SLE was characterized by a smaller female to male ratio (4.4:1 vs. 10.6:1; p = 3.10); a higher occurrence of serositis (36.7% vs. 28.6%; p = 7.10) and pulmonary involvement (21.2% vs. 11.3%; p = 6.10); and a lower occurrence of malar rash (31.1% vs. 62.4%; p = 10), photosensitivity (26.2% vs. 38.2%; p = 6.10), purpura/cutaneous vasculitis (13.4% vs. 25.9%; p = 9.10), alopecia/hair loss (24% vs. 44.9%; p = 3.10), Raynaud phenomenon (24.8% vs. 37.2%; p = 3.10), neuropsychiatric manifestations (15.3% vs. 20.2%; p = 0.025), lymphadenopathy (9.1% vs. 19.6%; p = 2.10), nephrotic syndrome (8.1% vs. 24.3%; p = 0.015), and nephritis (28.6% vs. 42.7%; p = 2.10). Regarding laboratory features, rheumatoid factor positivity was more frequent (32.7% vs. 20.1%; p = 3.10), whereas anti-RNP positivity (10.4% vs. 20.9%; p = 9.10), anti-Sm positivity (9.1% vs. 17.1%; p = 0.001), and a low CH50 complement fraction (45% vs. 64.9%; p = 0.002) were less frequent in old compared with young SLE patients. In conclusion, the clinical pattern of late-onset SLE is characterized by a lower disease severity. The reduced survival observed in this group seems to result mainly from the consequences of aging. PMID- 15525848 TI - The prognosis of acute respiratory failure in critically ill cancer patients. AB - Acute respiratory failure (ARF) in patients with cancer is frequently a fatal event. To identify factors associated with survival of cancer patients admitted to an intensive care unit (ICU) for ARF, we conducted a prospective 5-year observational study in a medical ICU in a teaching hospital in Paris, France. The patients were 203 cancer patients with ARF mainly due to infectious pneumonia (58%), but also noninfectious pneumonia (9%), congestive heart failure (12%), and no identifiable cause (21%). We measured clinical characteristics and ICU and hospital mortality rates.ICU mortality was 44.8% and hospital mortality was 47.8%. Noninvasive mechanical ventilation was used in 79 (39%) patients and conventional mechanical ventilation in 114 (56%), the mortality rates being 48.1% and 75.4%, respectively. Among the 14 patients with late noninvasive mechanical ventilation failure (>48 hours), only 1 survived. The mortality rate was 100% in the 19 noncardiac patients in whom conventional mechanical ventilation was started after 72 hours. By multivariable analysis, factors associated with increased mortality were documented invasive aspergillosis (odds ratio [OR], 2.13; 95% confidence intervals [CI], 1.05-14.74), no definite diagnosis (OR, 3.85; 95% CI, 1.26-11.70), vasopressors (OR, 3.19; 95% CI, 1.28-7.95), first-line conventional mechanical ventilation (OR, 8.75; 95% CI, 2.35-35.24), conventional mechanical ventilation after noninvasive mechanical ventilation failure (OR, 17.46; 95% CI, 5.04-60.52), and late noninvasive mechanical ventilation failure (OR, 10.64; 95% CI, 1.05-107.83). Hospital mortality was lower in patients with cardiac pulmonary edema (OR, 0.16; 95% CI, 0.03-0.72). Survival gains achieved in critically ill cancer patients in recent years extend to patients requiring ventilatory assistance. The impact of conventional mechanical ventilation on survival depends on the time from ICU admission to conventional mechanical ventilation and on the patient's response to noninvasive mechanical ventilation. PMID- 15525849 TI - Cardiovascular involvement, an overlooked feature of Erdheim-Chester disease: report of 6 new cases and a literature review. AB - Erdheim-Chester disease (ECD) is a rare, non-Langerhans form of histiocytosis of unknown origin. It is characterized by xanthomatous or xanthogranulomatous infiltration of tissues by spumous ("foamy") histiocytes. As of this writing, 178 cases have been reported. ECD is characterized by heterogeneous systemic manifestations. Bone pain is the most frequent symptom. About half of all patients have extraskeletal manifestations. Cardiovascular manifestations of ECD remain underestimated. We report 6 new cases of ECD associated with periaortic fibrosis. In 4 of these cases, the whole aorta had a "coated" aspect. A literature review revealed 66 cases of ECD with cardiovascular involvement. We therefore analyzed 72 ECD patients with cardiovascular involvement: 40 (55.6%) had periaortic "fibrosis," 32 (44.4%) had pericardial involvement, and 22 (30.6%) had myocardial involvement. Six had a right atrial tumor. Symptomatic valvular heart disease (3 aortic and 3 mitral regurgitations) was found in 6 patients. Nineteen patients (26.4%) had heart failure, leading to death in 8 cases. Six patients had renovascular hypertension related to perirenal artery stenosis. Data concerning follow-up were available for 58 (80.6%) patients. Of these, 35 (60.3%) patients died, confirming the severe prognosis of ECD. Cardiovascular complications were responsible for the death of 11 of the 35 patients (31.4%). PMID- 15525860 TI - Uncertainty intervention for watchful waiting in prostate cancer. AB - Watchful waiting is a reasonable alternative to treatment for some older men with localized prostate cancer, but it inevitably brings uncertainty. This study tested the effectiveness of the watchful waiting intervention (WWI) in helping men cognitively reframe and manage the uncertainty of watchful waiting. Based on Mishel's Reconceptualized Uncertainty in Illness Theory (Image. 1990; 256-262), the WWI was tested with a convenience sample of 41 men. Experimental subjects received 5 weekly intervention calls from a nurse. Control subjects received usual care. Outcomes were new view of life, mood state, quality of life, and cognitive reframing. Repeated measures of analysis of variance were used to test the effectiveness of the WWI. The sample was 86% Caucasian and 14% African American, with an average age of 75.4 years. Intervention subjects were significantly more likely than controls to view their lives in a new light (P = .02) and experience a decrease in confusion (P = .04) following the intervention. Additionally, intervention subjects reported greater improvement in their quality of life than did controls (P = .01) and believed their quality of life in the future would be better than did controls (P = .01). This study's findings document the benefits of the WWI for patients living with uncertainty. PMID- 15525861 TI - False-positive cancer screens and health-related quality of life. AB - By design, screening tests are imperfect-unresponsive to some cancers (false negatives) while occasionally raising suspicion of cancer where none exists (false positives). This pilot study describes patients' responses to having a false-positive screening test for cancer, and identifies screening effects on health-related quality of life (HRQoL). The pilot findings suggest issues important for incorporation in future evaluations of the impact of screening for prostate, lung, colon, or ovarian (PLCO) cancers. Seven focus groups were conducted to identify the nature and meaning of all phases of PLCO screening. Minnesota participants in the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial who had completed screening, with at least 1 false-positive screen, participated (N = 47). Participants' reactions to abnormal screens and diagnostic work-ups were primarily emotional (eg, anxiety and distress), not physical, and ultimately positive for the majority. Health distress and fear of cancer and death were the major negative aspects of HRQoL identified. These concepts are not typically included in generic HRQoL questionnaires like the SF 36, but are highly relevant to PLCO screening. Clinicians were regarded as underestimating the discomfort of follow-up diagnostic testing. However, relief and assurance appeared to eventually outweigh the negative emotions for most participants. Implications for oncology nurses include the need to consider the emotional consequences of screening in association with screen reliability and validity. PMID- 15525862 TI - Characteristics of participation in cervical cancer screening. AB - Although Papanicolaou test screening rates are reportedly high, a significant proportion of women remain unscreened. With recent revision of Papanicolaou test guidelines, it is critical that interventions and programs for cervical cancer directed toward low participating groups or individuals be developed. The purpose of the study was to examine factors that influence participation in cervical cancer screening by quantifying characteristics of women who engage in Papanicolaou test screening in a 12-month period. Using the 2000 National Health Interview Survey and Cancer Topical Module, the sample (N = 18,388) consisted of women who were older than 18 years. The dependent variable was nominally identified as whether a woman had had a Papanicolaou test in the last 12 months. Independent variables examined were insurance, level of education, place for care, age, race, employment, place of residence, and income level. Using logistic regression, all variables except race and income level were found to be significant for participation in cervical cancer screening (P < .000). Each variable is discussed within the framework of the Institute of Medicine model of access to personal healthcare services. Study findings provide insight and guidance for the development and implementation of methods for accessing women who have lower participation rates. PMID- 15525863 TI - Effects of 2 educational methods on the knowledge, attitude, and practice of women high school teachers in prevention of cervical cancer. AB - Because of the increased emphasis on prevention and early detection of cervical cancer, we studied the effects of 2 educational methods on the knowledge, attitude, and practice, as regards prevention of cervical cancer, of women high school teachers in Tabriz. This study was a semiexperimental research. Samples were 129 female teachers divided in 3 groups: experimental 1 (educated by pamphlets), experimental 2 (educated by a lecture and flash cards), and control group (not manipulated). After doing pretest in the 3 groups, investigators used 2 educational methods for experimental groups. Data regarding the knowledge and attitude of 3 groups were gathered after 14 days and data regarding practice were gathered after 2 months. Chi-square and 1-way ANOVA were used for data analysis. Before education, knowledge, attitude, and practice of the 3 groups were the same, but after education there were significant differences in mean scores of knowledge and attitude of 2 experimental groups as compared with the control group and also between the 2 experimental groups (P < .001). Education by lecture and flash cards was more effective than by pamphlets. In regard to Pap smear practice, there was a significant difference between the 2 experimental groups as compared with the control group (P = .001), but there was no significant difference between the 2 experimental groups. Therefore, educational methods were effective on knowledge, attitude, and practice of teachers regarding prevention of cervical cancer and education by lecture and flash cards was more effective than by pamphlets in increasing knowledge and inducing a positive attitude but the 2 educational methods had the same effect on practice of teachers. PMID- 15525864 TI - Health seeking related to ovarian cancer. AB - Critical review of general health-seeking models showed a need for expansion to include the early and atypical symptom period associated with ovarian cancer and the role of self and primary care in the diagnostic process. Data from family functioning research showed that in the self-care phase, the initial gastrointestinal symptoms were unrecognized as serious, given common sense labels, and self-managed. When primary care provider care was sought, misdiagnoses occurred three fourth of the time. Diagnostic delays occurred in these 2 phases of care. An expansion of a model of health seeking links personal and family risks and adds early symptom data may be obtained through monthly self monitoring by women using a symptom checklist. Organization of risks and symptom information assists in interpretation of disparate streams of data and gives a recommended outcome: high personal risk level + high family risk level + high early and persistent symptoms presence = high need for a prompt gynecological evaluation. The restructured health-seeking process requires women be taught how to monitor their ovarian health. Nurses and primary care providers need frequent continuing education updates and the health media need current and accurate information about this malignancy. PMID- 15525866 TI - Substantial changes in life: perceptions in patients with newly diagnosed advanced cancer and their spouses. AB - There are few studies on patients' perceptions of their situation after being recently diagnosed with an advanced gastrointestinal cancer and those of their spouses. Fourteen patients and their spouses were interviewed separately. The interviews were analyzed using a phenomenographic approach. The analysis indicated that the response categories for patients and spouses were roughly the same, but the number of patients and spouses who made statements differed between categories. All informants perceived substantial changes in life. This included negative physical, mental, and practical changes as well as positive changes. Mental changes included 3 categories: despair, why, and uncertainty. The informants described several ways of handling these changes in life. The most frequently reported by patients were that "one shouldn't complain" and by spouses to "hope," and by all informants to "make the best of it." Other ways of handling the situation were reconciliation, avoidance, preparation for death, seeking support, and isolation. In conclusion, more patients than spouses seemed to accept their situation because fewer patients complained and instead prepared for death, whereas more spouses felt despair, used hope and avoidance, and were preoccupied with practical matters. These findings suggest that spouses are a vulnerable group and healthcare staff should be just as aware of their situation as that of the patients. PMID- 15525867 TI - Symptoms, psychological distress, social support, and quality of life of Chinese patients newly diagnosed with gastrointestinal cancer. AB - This study aims to describe symptoms, psychological distress, social support, and quality of life of Chinese patients newly diagnosed with gastrointestinal tract (GIT) cancer, and to identify the extent to which demographic, physical, and psychosocial factors predict their quality of life. A convenience sample of 146 newly diagnosed GIT cancer patients recruited from 3 major hospitals in Shanghai completed a self-report questionnaire. The questionnaire was designed to obtain demographic and medical data and measures of symptoms, psychological distress, social support, health-related quality of life (HRQoL), and global quality of life (GQoL). Measures developed in English were translated into Chinese using the procedure advocated by WHO. The results showed that the most common signs and symptoms reported were fatigue, pain, and weight loss; 28% of the patients were depressed; and overall, patients had a moderate quality of life. Comparative analyses found some difference on measures for demographic and diagnostic subgroups. Depression, symptom distress, and social support accounted for 44% of the total variance for HRQoL, while perceived financial difficulty and symptom distress accounted for 20% of the total variance for GQoL. Findings from this research give insights into the importance of quality of life assessment, symptom management, and intervention to improve the quality of life of Chinese cancer patients. It also raises questions about measures of quality of life that are culturally relevant. PMID- 15525868 TI - Determining the main risk factors and high-risk groups of breast cancer using a predictive model for breast cancer risk assessment in South Korea. AB - This study was aimed at developing a predictive model for assessing the breast cancer risk of Korean women under the assumption of differences in the risk factors between Westerners and Koreans. The cohort comprised 384 breast cancer patients and 2 control groups: one comprising 166 hospitalized patients and the other comprising 104 nurses and teachers. Two initial models were produced by comparing cases and the 2 control groups, and the final equations were established by selecting highly significant variables of the initial models to test the accuracy of the models in terms of disease probability and predictability. Both the initial models and the final disease-probability models were confirmed to exhibit high degrees of accuracy and predictability. Major risk factors determined by comparing the patients with hospitalized controls were a family history, menstrual regularity, total menstrual duration, age at first full term pregnancy, and duration of breastfeeding. Major risk factors determined by comparing patients with nurse/teacher controls were age, education level, menstrual regularity, drinking status, and smoking status. The predictive model developed here shows that risk factors for breast cancer differ between Korean and Western subjects in the aspect of breastfeeding behavior. However, identifying the relationship between genetic susceptibility and breast cancer will require further studies with larger samples. In a model with nurse/teacher controls, drinking and higher education were found to be protective variables, whereas smoking was a risk factor. Hence the predictive model in this group could not be generalized to the Korean population; instead, breast cancer incidence needs to be compared among nurses and teachers in a nurse-and-teacher cohort. PMID- 15525869 TI - Breast cancer knowledge assessment in female Chinese immigrants in New York. AB - The purpose of this study was to examine the relationships between acculturation level and perceptions of health access, Chinese health beliefs, Chinese health practices, and knowledge of breast cancer risk. This descriptive, correlational cross-sectional study used a survey approach. The sample included 135 Chinese women from the New York City metropolitan area. Data were analyzed using correlational techniques and polytomous regression. There were no significant relationships between acculturation and health access, Chinese health beliefs, Chinese health practices, and breast cancer risk knowledge. Only "years of education," "marital status," and "household income" significantly predicted breast cancer risk knowledge level. The data indicate that women with a better knowledge of breast cancer risk are twice as likely to have higher income and have more education. The most knowledgeable women are less likely to be married and less likely to have partners compared to least knowledgeable group. Providers need to promote health knowledge and provide information about as well as access to preventive health practices to the immigrant population, given that acculturation to the new dominant society is inevitable. PMID- 15525870 TI - Competence of and emotional/behavioral problems in pediatric oncology patients in Taiwan. AB - The objective of this study was to examine the factor structure of Achenbach's Child Behavior Checklist, using confirmatory factor analysis, in a group of pediatric oncology patients in Taiwan and to compare the competence and behavioral/emotional problems scores of these children with normative scores to assess competence and emotional/behavioral problems of Taiwanese children with cancer. Subjects included parents (124 mothers and 22 fathers) of 146 pediatric oncology patients who completed the measures. Factor loadings of the overall fit indices for the study subjects yielded or approached satisfactory results for all syndromes, except for aggressive behavior. The indices for both internalizing and externalizing syndromes also showed good model fit. Taiwanese children with cancer had significantly lower competence scores on the Child Behavior Checklist/4-18 than did Achenbach's sample, except Taiwanese girls with cancer, who had higher activity competence scores, but these scores were not statistically significant. Taiwanese boys with cancer had significantly higher scores on all of the emotional/behavioral problems, but had lower delinquent behavior, aggressive behavior, and externalizing syndrome scores. In contrast, girls with cancer had significantly higher scores on somatic complaints and internalizing syndrome, but significantly lower aggressive behavior and externalizing syndrome scores. Taiwanese adolescents with cancer had significantly lower competence scores compared with normative scores, with the exception being that the adolescents in the current study had significantly higher school competence scores. These adolescents also had significantly fewer attention problems but tended to have more somatic complaints. These findings clearly indicate that healthcare for children with cancer should include psychological services to prevent long-term emotional/behavioral problems. PMID- 15525872 TI - Metabolism of lipids in human white adipocyte. AB - Adipose tissue is considered as the body's largest storage organ for energy in the form of triacylglycerols, which are mobilized through lipolysis process, to provide fuel to other organs and to deliver substrates to liver for gluconeogenesis (glycerol) and lipoprotein synthesis (free fatty acids). The release of glycerol and free fatty acids from human adipose tissue is mainly dependent on hormone-sensitive lipase which is intensively regulated by hormones and agents, such as insulin (inhibition of lipolysis) and catecholamines (stimulation of lipolysis). A special attention is paid to the recently discovered perilipins which could regulate the activity of the lipase hormono sensible. Most of the plasma triacylglycerols are provided by dietary lipids, secreted from the intestine in the form of chylomicron or from the liver in the form of VLDL. Released into circulation as non-esterified fatty acids by lipoprotein lipase, those are taken up by adipose tissue via specific plasma fatty acid transporters (CD36, FATP, FABPpm) and used for triacylglycerol synthesis. A small part of triacylglycerols is synthesized into adipocytes from carbohydrates (lipogenesis) but its regulation is still debated in human. Physiological factors such as dieting/fasting regulate all these metabolic pathways, which are also modified in pathological conditions e.g. obesity. PMID- 15525873 TI - Aldosterone and anti-aldosterone effects in cardiovascular diseases and diabetic nephropathy. AB - Cells in the cortical collecting duct of distal nephron have been considered for a long time as the unique cellular targets of aldosterone. However, it is now clear that other cell types in non-epithelial tissues are also potential targets for aldosterone. The functions that this hormone controls in non-epithelial tissues are still a matter of debate. Clinical and experimental studies have established that aldosterone plays a major role in the pathophysiology of cardiovascular and renal diseases. The aldosterone receptor antagonists spironolactone and eplerenone have demonstrated specific effects not related to their hypotensive properties in hypertension or cardiac diseases. It appears that a key action of these molecules is related to prevention or treatment of end organ damage. The latter fact, and the recognition of aldosterone escape on long term treatment of heart failure, diabetic nephropathy and some forms of hypertension with ACE inhibitors, justify the clinical use of aldosterone receptor antagonists provided that kaliemia is controlled. Experimental studies have allowed to draw a still incomplete but comprehensive scheme of aldosterone cardiovascular actions in pathological conditions. When elevated, aldosterone has deleterious effects in blood vessels, in the heart and in kidney, which are secondary to the induction of inflammatory and oxidative processes and necrosis, that induce the increased synthesis of extracellular matrix proteins. PMID- 15525874 TI - Dexamethasone-induced insulin resistance shows no gender difference in healthy humans. AB - OBJECTIVE: Recent reports suggest that lipid-induced insulin resistance is more pronounced in men than in women. Whether such gender difference exists for other factors known to induce insulin resistance in healthy individuals remains unknown. We therefore assessed whether glucocorticoid-induced insulin resistance differs in men and women. METHODS: The insulin sensitivity and insulin secretion of 8 women and 7 men, all non obese and healthy, were evaluated with or without administration of dexamethasone (2 mg/day during 2 days) by means of a two-step hyperglycemic clamp. RESULTS: Dexamethasone decreased insulin sensitivity to the same extent in men and women. The relative increases in insulin concentration observed after dexamethasone in the basal state, during the first phase of insulin release and at the two steps of hyperglycemia were similar in men and women. The hyperinsulinemia thus attained allowed to fully compensate for insulin resistance in both genders. CONCLUSIONS: The effects of glucocorticoids on insulin sensitivity and insulin secretion show no gender difference in healthy humans. PMID- 15525875 TI - Sub-optimal drug treatment of diabetes and cardiovascular risk in diabetic patients in Turkey. A countrywide survey. AB - OBJECTIVES: The present study is a snapshot of how diabetic patients are treated for diabetes and coexisting cardiovascular risk factors in Turkey. We also addressed the question of what percentage of these patients are treated appropriately according to the current guidelines. Next step will be to determine which pharmacological treatment strategies affect mortality and morbidity in these patients and whether there are regional differences in these outcomes. METHODS: To get a representative picture, Turkey was splitted into four parts with different ethnic and socioeconomic features then centers were randomized within each of these parts. Number of the centers in a region were calculated according to the population of that region. 305 physicians in 11 cities participated in data collection during a period of 3 months. Consecutive 2226 diabetic patients patients who were above 55 years of age were included. Detailed information was obtained about the demographic features and the cardiovascular risk factor and diabetes status of the patients together with relevant drug treatment. Laboratory analyses were done locally and recorded if performed during the last 3 months. RESULTS: Most patients were treated with oral antidiabetic monotherapy regardless of diabetes duration, metabolic control and complication and cardiovascular risk factor status. There was a trend among physicians except for endocrinologists to underprescribe insulin. Monotherapy also was the main mode of treatment for hypertension. Angiotensin converting enzyme inhibitors were generally not used as first line treatment contrary to the recommendations and angiotensin converting enzyme inhibitors and angiotensin receptor blockers are not prescribed for renoprotection in microalbuminuric patients. Statins, fibrates, metformin and aspirin were largely underused. CONCLUSION: The present study indicates that diabetic patients are undertreated in Turkey. Therefore every effort should be spent to implement current guidelines in diabetic patients in order to prevent macro and microvascular complications of diabetes. PMID- 15525876 TI - Prevalence of diabetes, obesity, hypertension and hyperlipidemia in the central area of Argentina. AB - OBJECTIVES: There is little information available about the prevalence of chronic metabolic diseases in many Latin American countries. Between 1995 and 1998, studies on the prevalence of obesity, hypertension, hyperlipidemia and diabetes were carried out in four cities located in central Argentina: Dean Funes, Oncativo, Pehuajo and Venado Tuerto. The data provided by these surveys are reanalysed here in order to determine prevalence of obesity, hypertension, hyperlipidemia and diabetes using new epidemiological criteria. METHODS: Representative samples of the population, based on a multistage probabilistic sampling design, were taken from each of the four cities. The sample size was calculated to obtain a precision of 4% for the prevalence assessment. The subjects included were aged 20 years and over. Standardization of the prevalence rates used the entire study sample as the reference population. RESULTS: Age standardised prevalence rates for the cities ranged between 22.4% and 30.8% for obesity, 27.9% and 43.6% for hypertension, 24.2% and 36.4% for hyperlipidemia, and 6.5% and 7.7% for diabetes mellitus. All these prevalences increased with age. 58.1% of the obese subjects and 51.2% of the diabetic subjects had hypertension, while 43.2% of the obese subjects and 52.8% of the diabetic subjects had hyperlipidemia. CONCLUSIONS: While the prevalence of diabetes mellitus was between 6% and 8%, the prevalence of obesity was close to 26% and hypertension and hyperlipidemia affected one third of the population. These data can be considered as indicative of the prevalences of these four diseases in the population aged 20 years and over, in the central region of Argentina. PMID- 15525877 TI - Intravitreous levels of hepatocyte growth factor/scatter factor and vascular cell adhesion molecule-1 in the vitreous fluid of diabetic patients with proliferative retinopathy. AB - OBJECTIVE: Hepatocyte growth factor, also know as the scatter factor (HGF/SF) has been involved in the etiopathogenesis of proliferative diabetic retinopathy (PDR). To further explore this issue we have determine the intravitreous levels of HGF/SF taking into account the problems that could lead to misinterpretation of the results when the vitreous fluid is used to indirectly explore the events that are taking place in the retina. In addition, the relationship between HGF/SF and vascular cell adhesion molecule 1 (VCAM-1) was also investigated. PATIENTS AND METHODS: Serum and vitreous samples were obtained during vitrectomy from 22 diabetic patients with PDR and 25 non-diabetic control subjects. Patients in whom intravitreous hemoglobin was detectable were excluded. A correction for plasma levels of either HGF/SF and VCAM-1 and intravitreal proteins was performed. RESULTS: Vitreal levels of both HGF/SF and VCAM-1 were higher in patients with PDR in comparison with the control group (p<0.001 and p=0.003, respectively). However, after correcting for total vitreal proteins both HGF/SF and VCAM-1 (ng/mg of proteins) were lower in diabetic patients than in non-diabetic control subjects (p=0.03 and p<0.0001, respectively). No relationship between the vitreous levels of either HGF/SF or VCAM-1 with PDR activity was detected. Finally, a correlation between the vitreal levels of HGF/SF and VCAM-1 was observed in diabetic patients (r=0.61, p=0.005) but not in the control group. CONCLUSION: Our results suggest that intraocular production of HGF/SF might be more important in mediating inflammatory and fibroproliferative processes rather than in angiogenesis itself. PMID- 15525878 TI - C677T methylene-tetrahydrofolate reductase mutation in type 2 diabetic patients with and without hyperhomocysteinaemia. AB - OBJECTIVE: The aim of the study was to determine the prevalence of the C677T mutation in a cohort of type 2 diabetic patients with and without elevated total plasma homocysteine (tHcy). METHODS: 80 type 2 diabetic patients with hyperhomocysteinaemia (group 1, tHcy: 21.3 +/- 6.7 micromol/L) and 50 subjects with normal levels (group 2, tHcy 11.2 +/- 2.3 micromol/L) were studied. C677T mutation was assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Homozygosity was present in 23% of patients in group 1 and 8% in group 2 (P<0.02). No significant difference in heterozygosity frequency was observed between patients with and without hyperhomocysteinaemia. T allele frequency was 0.43 in group 1 and 0.35 in group 2. CONCLUSION: C677T mutation is frequent in diabetic patients with hyperhomocysteinaemia and could contribute, besides non genetic factors, to increased levels of tHcy. PMID- 15525879 TI - Clinical significance of alendronate in postmenopausal type 2 diabetes mellitus. AB - OBJECTIVES: To examine early changes in biochemical markers of bone turnover and bone mineral density (BMD) in a clinical trial of anti-resorptive agent alendronate versus alfacalcidol in postmenopausal women with type 2 diabetes mellitus. METHODS: 12 subjects (mean age; 73.1 +/- 6.3 yrs, duration of diabetes; 13.2 +/- 3.7 yrs) were administered alendronate sodium (5 mg/day) and 12 subjects (mean age; 70.7 +/- 7.8 yrs, duration of diabetes; 12.8 +/- 2.0 yrs) were administered alfacalcidol (0.5 microg/day) for 12 months. Urinary N-telopeptide cross-linked collagen type I (NTx), one of biochemical markers, and radial bone mineral density (BMD) were measured as a marker of bone turnover. RESULTS: After 12 months, urinary NTx did not change in women with alfacalcidol treatment, however urinary NTx significantly decreased after alendronate treatment. The BMD significantly decreased by 3.33% (p<0.05) in women with alfacalcidol treatment, while the BMD did not decrease in women with alendronate treatment. CONCLUSION: Alendronate that produces reduction in urinary NTx and inhibition of decrease in BMD may have a clinical significance to reduce the risk of bone fracture in postmenopausal type 2 diabetic women. PMID- 15525880 TI - Parent-child physical activity relationships in 12-year old French students do not depend on family socioeconomic status. AB - OBJECTIVES: Regular physical activity (PA) in young needs to be promoted to prevent obesity and subsequent diabetes. Influences of parental PA on adolescents' activity level have been studied with discordant results and the effect of socioeconomic status (SES) on these relationships is not clarified. METHODS: Cross-sectional population-based survey of about 3000 12-year old French students and their parents. Familial associations were assessed with logistic regression models taking into account SES and children's and parents' corpulence. Sport involvement and sedentary behaviors were assessed by standardized questionnaires filled out separately by children and parents, the latter also reporting their educational data and family income tax. RESULTS: Participation in structured PA outside school was higher for boys (74%) than for girls (58%, p<10 5) and a high sedentary behavior (watching television, playing computer/video games and reading > or =2h/day) was observed in one-third of the sample, both in boys and in girls. Children were more likely to participate in structured PA outside school when both parents practiced sport as compared to neither parent practicing it, with an odd ratio OR (95%CI) of 1.97 (1.4-2.8) for boys and 1.56 (1.2-2.1) for girls. Familial associations of inactivity were significant for boys only. A greater percentage of adolescents had a high level of sedentary behavior when both parents versus no parents watched television > 2h/day (OR 1.95 (1.52.6)). Parent-child physical activity relationships were not modified by the family SES or the children's or parents' weight status. CONCLUSION: Parental involvement in sport is an important correlate of a young adolescent's participation in structured PA outside school, whatever the family SES or corpulence of the family members. PMID- 15525881 TI - Maternal-fetal transport kinetics of L-Leucine in vitro in gestational diabetic pregnancies. AB - OBJECTIVE: Paucity of data relating to transport of amino acids in gestational diabetic pregnancies prompted us to undertake this study. Transport kinetics of a model amino acid, L-leucine was investigated in gestational diabetic pregnancies in vitro, using perfusion of isolated placental lobules. METHODS: Placentae from diabetic and control pregnant women were collected post-partum. Suitable placental lobules were then perfused, using National Culture and Tissue Collection (NCTC) medium, diluted with Earle's buffered salt solution as perfusate. 14C-labelled L-leucine along with tritiated water as reference were injected as a 100 ul bolus into the maternal circulation and serial perfusate samples collected over a 5-minute study period. RESULTS: In 6 successful perfusions, differential transport rate of L-leucine for 10, 25, 50, 75 and 90% of efflux in the fetal vein averaged 1.17, 1.12, 1.22, 1.20 and 1.17 times respectively that of reference in the diabetic group. In the control group (n=6), leucine transport indices for the corresponding efflux periods averaged 1.13, 1.15, 1.18, 1.17 and 1.16 times respectively that of the reference marker. Student's 't' test showed that the difference between the two groups was not statistically significant (p > 0.05) for all the efflux fractions studied. In the diabetic series, leucine transport fraction (TF) averaged 41.2 +/- 4.5% of corresponding water TF while in control group, the amino acid TF averaged 46.5 +/ 6.5% of water TF. The difference between the two series, however was not statistically significant (p > 0.05). Similarly, kinetic parameters as area under the curve, clearance, elimination constant, time for maximum response, absorption rate, and elimination rate in the diabetic and control groups, were not significantly different (p > 0.05) as well. CONCLUSION: Our study seems to indicate that transport kinetics of leucine under in vitro conditions, do not differ significantly in placentae of gestational diabetic women compared to controls. PMID- 15525882 TI - Emphysematous cystitis. AB - We present a case of emphysematous cystitis in a diabetic patient with a poor glycemic control in the context of alcoholic chronic pancreatitis. A 62-year-old woman was admitted to the emergency department after being found on floor with confusion and vomiting. The clinical examination was unremarkable except she was undernourished, agitated and presented an hepatomegaly. Urine contained 5.104 leukocytes/mm3 and culture grew Escherichia coli, 10(7) Colony Forming Unit/ml. Abdominal plain film showed gas shadows along the wall of urinary bladder. CT scan of the pelvis confirmed the presence of gas, and diffuse thickening of the urinary bladder wall. A Foley catheter was placed and the patient was treated with antibiotics for 6 weeks. She was also treated with insulin, rehydratation, vitamin B1 and B6, and pancreatic enzyme replacement. Emphysematous cystitis is defined by the presence of gas in the urinary bladder wall. It complicates urinary tract infections especially in diabetic patients but other disabled general medical conditions may be present. Because this relatively uncommon disease may present with fairly nonspecific findings, the diagnosis is often made incidentally on X-rays. However, as early diagnosis and treatment improve the outcome, a high index of suspicion for unusual presentations is warranted. Every diabetic patient with a urinary tract infection who seems to be severely ill should have an abdominal X-ray as a minimal screening tool to detect emphysematous complications. PMID- 15525883 TI - Usefulness of B-type natriuretic peptide (BNP) as a screen for left ventricular abnormalities in diabetes mellitus. AB - Cardiovascular disease is the leading cause of morbidity and mortality in diabetic subjects. Diabetes, independently of the mechanism, is associated with an increased risk of left ventricular hypertrophy, left ventricular dysfunction and coronary artery disease. B-type natriuretic peptide (BNP) is a cardiac neurohormone predominantly released from the cardiac ventricles in response to left ventricular volume expansion and pressure overload. Numerous studies have shown that BNP levels are elevated in asymptomatic or symptomatic left ventricular dysfunction, hypertrophy and coronary artery disease. BNP testing plays an important role in the screening and diagnosis of left ventricular dysfunction by improving the performance of non-specialist physicians in diagnosing heart failure. In clinical practice, BNP testing is best used as a 'rule out' test targeted to patients at high risk for left-ventricular dysfunction, such as those with diabetes. Studies are needed to establish if this promising biological tool, in the next future, would assist the management of diabetic patients. PMID- 15525884 TI - Ethical, deontologic, social, and economic reflections on screening with helical CT. Part I: general aspects. PMID- 15525885 TI - Ethic, deontologic, social, and economic reflections on screening with helical CT. Part II: specific aspects. PMID- 15525886 TI - Role of the chest radiograph in the preoperative assessment of the pulmonary function in patients with cirrhosis candidates to liver transplant. AB - PURPOSE: To appraise the role of chest radiography (CXR) in the clinical assessment of pulmonary complications occurring as a result of liver disease of any cause, a standardised reading of CXR was compared both with the results of lung function studies and with the severity of liver disease in 60 consecutive patients with cirrhosis candidates to liver transplant. MATERIALS AND METHODS: CXR were scored for signs of cardiomegaly, enlargement of central and or peripheral vessels, hyperkinetic circulatory and/or intravascular volume states, abnormally increased interstitial lung markings, and pleural effusion by three independent observers. RESULTS: The CXR score of this complete vascular interstitial deficiency showed a high interobserver reproducibility and was significantly increased in those patients with a more decompensated liver cirrhosis. Moreover, a larger number of CXR signs of vascular and interstitial deficiency was present in those patients who had a more severe cirrhosis (Child Pugh class 3). Functional indices of pulmonary gas exchange and diffusing capacity were worse in patients with end-stage liver cirrhosis, and for some indices the correlation was statistically significant. The comparison between lung function and CXR score showed that on average lung function impairment was significantly less severe in patients with lower CXR scores. CONCLUSIONS: In conclusion, the information derived from a standardised reading of CXR is valuable in the clinical assessment of the pulmonary damage induced by liver cirrhosis as it allows discrimination of classes of severity of liver disease and of lung function impairment. When integrated with physiologic measurements and, if necessary, with high resolution tomographic studies, chest radiography helps to discriminate patients with advanced liver cirrhosis and progressing towards a hepatopulmonary syndrome, who need a proper therapeutic management. PMID- 15525887 TI - Advantages and limits of percutaneous breast core biopsy with Mammotome and stereotactic equipment in upright seated patient. AB - PURPOSE: To confirm the diagnostic accuracy of percutaneous core breast biopsy with Mammotome and stereotactic equipment in upright seated patients, and to compare results from this method with those obtained using the dedicated unit with prone table. MATERIALS AND METHODS: We analysed 72 microbioptic samples from 69 upright seated patients. These samples were obtained in the course of mammographies assisted by digital stereotactic equipment (Diamond Delta32 General Electric Medical System/Instrumen-tarium), and further equipped with a lateral arm to support the Mammotone probe. We studied the following lesions: 57 cases of isolated microcalcification clusters, 12 masses with microcalcification and three cases of plain masses. We describe in detail the various phases of our 30-minute long microbiopsy procedure. All 72 samples were extracted without problems. Only in one case a large haematoma emerged, but quickly disappeared. RESULTS: We were able to characterise the histology of the lesions in all our samples: in 39/72 cases they were identified as benign, 28 were classified as malignant, and the remaining 5 as atypia. In the 39 lesions identified as benign, the microbiopsy procedure could be carried out without surgical biopsy - consequently, these patients were available for routine follow-up exams, which confirmed the benign nature of their lesions. Our identification of the 28 malignant lesions was confirmed by surgical biopsy. The microbiopsy finding and the histological result did not agree in only two cases. The 5 cases with atypical ductal hyperplasia showed no neoplastic alteration following surgical biopsy. In our personal experience, core biopsy with Mammotome achieved a high diagnostic accuracy. CONCLUSIONS: After careful analysis of the results obtained using Mammotome in upright seated patients with stereotactic equipment, we can confirm the diagnostic accuracy of this procedure in mammography alterations not visible in sonography. In agreement with previous studies, we also confirm that the diagnostic accuracy of this approach, as well as possible complications deriving from it, do not depend on the position of the patient. PMID- 15525888 TI - Role of ultrasound-guided fine needle cytology of axillary lymph nodes in breast carcinoma staging. AB - PURPOSE: To evaluate the efficacy of cytology on axillary lymph node ultrasound guided aspiration biopsy in the reduction of inappropriate surgery, such as the sentinel node (SN) procedure if positive, or axillary dissection if negative. MATERIALS AND METHODS: Cytology was performed on 159 consecutive cases, on the ultrasonographically most suspicious lymph node. Lymph node histology was used as a reference standard to determine accuracy. Four different scenarios were simulated: routine axillary ultrasonography with cytology of the lymph nodes visible at ultrasonography (A), or of only the lymph nodes suspicious at ultrasonography (B), with ultrasonography limited to clinically negative axillae and cytology of the lymph nodes visible at ultrasonography (C), or only of the lymph nodes suspicious at ultrasonography only (D). RESULTS: Cytologic sensitivity was 58.6%, specificity 100%. Immediate axillary dissection only in the case of positive cytology would have avoided 6/6 inappropriate axillary dissections and 5/34 (14.7%) inappropriate SN, compared to routine practice (immediate dissection for palpable adenopathy, SN in the remaining cases). Each of the simulated scenarios saved inappropriate surgical procedures (A: 6 dissections, 5 SNs; B: 6 dissections, 3 SNs; C: 13 SNs; D: 11 SNs) at no expense (A and B) or limited expense (C: Euro 348, D: Euro 232 for each inappropriate surgical procedure saved). CONCLUSIONS: Axillary lymph node cytology can save axillary dissections or sentinel node procedures and is recommended as routine practice. Routine axillary ultrasonography, with cytology of sonographically visible lymph nodes, followed by immediate axillary dissection only in case of positive cytology proved to be the best approach in terms of cost-benefit ratio. PMID- 15525889 TI - Treatment of large hepatocellular carcinoma: comparison between techniques and long term results. AB - PURPOSE: The aim of this study was to compare results over time of Transcatheter Arterial Chemo Embolization (TACE), Percutaneous Ethanol Injection (PEI), Laser Thermal Ablation (LTA) and combined therapy of large Hepatocellular Carcinoma (HCC). MATERIALS AND METHODS: Between 1995 and 2003, 89 cirrhosis patients (51 Child-Pugh A, 38 Child-Pugh B) with at least one nodule of HCC =/> 40 mm, were included in this randomized study; 21 were treated with TACE, 20 with PEI, 29 with LTA and 19 with combined therapy. The total number of HCC nodules was 92 with a mean diameter of 52.9 mm. RESULTS: No major complication occurred in all procedures. CT scan showed that complete necrosis was achieved in 83% of treated nodules (76 out of 92); as a whole, the disease relapsed in 18 (20.2%) patients (disease free interval being 18.2+/-9.4 months). The cumulative survival rates were 69.6%, 25.1% and 9.8% at 12, 36 and 60 months respectively. Univariate analysis of survival showed statistically significant differences in the comparison between Child-Pugh A group vs B (p<0.0001) and between single nodule vs multiple (p=0.0019). Patients subjected to combined therapy and LTA showed a statistically significant longer survival than those treated with TACE and PEI. CONCLUSIONS: LTA proves to be the most effective treatment for HCC < 50 mm, combined therapy is the best choice for nodules =/> 50 mm since complete necrosis is achieved in almost all cases and better total survival in the treated patients. PMID- 15525890 TI - Multislice CT anatomy of hepatic artery in patients undergoing liver transplantation using 3D reconstructions. AB - PURPOSE: To evaluate the accuracy of multislice CT angiography, investigating vascular anatomy and anatomical variants of hepatic artery in patients undergoing liver transplantation. MATERIALS AND METHODS: The study concerns 20 patients (12 male and 8 female) candidates to liver transplantation were examined using multislice CT with a triphasic protocol following the administration of Iomeron 400 mg/ml at a rate of 5 ml/s using Sure Start technique. The following protocol was applied in all patients: row thickness 3, pitch 5.5, image thickness 3, reconstruction 1. Vascular reconstruction was obtained with 3D Maximum Intensity Projection and Volume Rendering algorithms using the data of the arterial phase. All variants were classified by Michels's classification. All patients were transplanted and the anatomical results of CT have been verified surgically. RESULTS: CT angiography detected 5 anatomical variants of the hepatic artery and one aneurysm of the celiac trunk; the other 14 patients had a normal anatomy. In all patients the results of CT correlates with the surgical ones. CONCLUSIONS: Our results suggest that multislice CT angiography is useful for planning surgical transplantation, giving precise information about vascular anatomy and its variants, (those are common). In our opinion this technique can replace conventional angiography. PMID- 15525892 TI - Occlusion of postcatheterisation femoral pseudoaneurysms with percutaneous thrombin injection under ultrasound guidance. AB - PURPOSE: The incidence of iatrogenic femoral artery pseudoaneurysms is reported to occur in 1-7% by of all percutaneous catheterisations. These pseudoaneurysms are traditionally treated by ultrasound-guided compression or surgical repair. We report our experience in sealing postcatheterization femoral pseudoaneurysms with percutaneous thrombin injection under colour-Doppler ultrasound guidance. MATERIALS AND METHODS: Since June 2000 we have consecutively treated 31 pseudoaneurysms in 30 patients, (14 males and 16 females, age range 45 to 81 years); in one patient the pseudoaneurysm was bilateral. All patients had a clinical diagnosis of postcatheterization femoral pseudoaneurysm, later confirmed by colour-Doppler ultrasonography. We injected a bovine thrombin solution percutaneously at a concentration of 1000 U/mL using 21-22 gauge needles under colour Doppler ultrasound guidance. All patients underwent clinical and colour Doppler US examination before, during and 24 hours after the procedure and were followed up after 1 and 3 months. RESULTS: The primary success rate was 83.8%. Complete and persistent occlusion of the pseudoaneurysm was achieved in less than 20 seconds by administering an average dose of 880 U of thrombin (0.8 mL of solution). In 5 cases (16.1%) reperfusion of the pseudoaneurysm was observed within 24 hours. These patients underwent a repeat procedure. The final result was successful in 96.7% of patients (30 of 31 cases). No thromboembolic complication was observed. Only 22.5% of patients reported a heat sensation in the treated limb, which resolved spontaneously within minutes. CONCLUSIONS: The percutaneous injection of thrombin under ultrasound colour-Doppler guidance should be regarded as the first choice treatment for postcatheterization femoral pseudoaneurysms, owing to its simplicity, safety, effectiveness and inexpensiveness. PMID- 15525891 TI - Ultrasound diagnosis of intragastric balloon complications in obese patients. AB - PURPOSE: Our study describes ultrasonography (US) aspects of complications related to the use of the Bioenterics intragastric balloon (BIB) system for the treatment of moderate obesity. MATERIALS AND METHODS: Over the last three years 151 BIBs were endoscopically placed and inflated with 500-600 ml of saline plus 10 ml of methylene blue in 131 patients (mean age 38.4 years, range 18-72; 46 males, 85 females, mean body mass index 43.8 kg/m(2)). In all cases abdominal US was carried out before and 1-2 days after endoscopy and thereafter at scheduled follow-up visits and/or whenever the patient complained of heartburn, vomiting, a sensation of ''absence of the BIB'' with or without staining of urine and/or faeces. RESULTS: The BIB appeared as a round anechoic structure, with a thick hyperechoic wall and a hyperechoic band-like valve inside. In 8 cases staining of urine and/or faeces prompted immediate endoscopic removal. In 10 patients US findings of BIB-related complications were: 1) decreased volume with a visible valve (5 cases; in two of these staining of urine was reported); 2) multiple hyperechoic streaks regularly spaced due to a completely collapsed BIB, not modified by decubitus changes (3 cases; one patient reported staining of urine); 3) migration through the bowel (2 cases): in one patient the BIB was passed after 4 days, whereas in the other case bowel obstruction required laparoscopic surgery. CONCLUSIONS: In conclusion, US provides a quick and accurate method for the assessment of BIB-related complications in obese patients. PMID- 15525893 TI - Computed tomography and magnetic resonance features of carcinoma of the tongue. AB - Computed tomography (CT) and magnetic resonance (MR) imaging allow more accurate staging of the carcinoma of the tongue than the clinical examination alone. The complex anatomy and the tumour margins make the interpretation of CT and MR images challenging. The aim of this paper is to review the technique and semiotic criteria for CT and MR evaluation and staging of carcinoma of the tongue. PMID- 15525894 TI - Magnetic resonance imaging of the upper airway in obstructive sleep apnea. AB - PURPOSE: To define the role and reliability of MR imaging of the oropharynx during wakefulness in the evaluation of patients with Obstructive Sleep Apnoea Syndrome. MATERIALS AND METHODS: During a 33-month period, 28 patients with clinical diagnosis of Obstructive Sleep Apnoea Syndrome (OSAS) were evaluated with Magnetic Resonance imaging. MR imaging was performed during wakefulness by T1-, DP- and T2-weighted sequences in the sagittal and axial plane. The morphological and quantitative data of the imaging, the majority of which were obtained from the relevant Literature (minimal pharyngeal cross-sectional area and its location and shape, increase of adipose tissue of the neck, tongue swelling, posterior dislocation of the tongue, soft and hard palate length, geniohyoid muscle length, evaluation of oral floor lowering), were subsequently compared with clinical-instrumental parameters (Body Mass Index, apnoea/hypopnoea index, hematocrit, arterial oxygen partial pressure, average nocturnal oxyhemoglobin saturation and sleeping-time percentage with oxygen saturation lower than 90%), to assess correlations. RESULTS: The minimal pharyngeal cross area is significantly positively correlated with the arterial oxygen partial pressure and with the average nocturnal oxyhemoglobin saturation and is inversely correlated with the apnoea/hypopnoea index and with the sleeping-time percentage with oxygen saturation lower than 90%; the inverse correlation with the Body Mass Index is less marked. Furthermore, the minimal pharyngeal cross-sectional area is correlated with its shape on the axial section. Increase of the adipose tissue of the neck is significantly correlated with the Body Mass Index, with a low nocturnal oxygen saturation, with a high percentage of sleeping time with oxygen saturation lower than 90%, and with the axial-section shape of the minimal pharyngeal area. An inverse correlation between the soft palate length and the arterial oxygen partial pressure was observed. Oral floor lowering is significantly inversely correlated with the minimal area only if the minimal area is located at the tongue base level and is directly correlated with the Body Mass Index as well as with the apnoea/hypopnoea index. The remaining MR findings appear to be independent parameters that may worsen the syndrome when associated with other parameters. CONCLUSIONS: In patients with a clinical diagnosis of OSAS, MR imaging performed during wakefulness can provide reliable information on the causes, severity and site/s of oropharyngeal obstruction with important consequences on therapeutic decisions. PMID- 15525895 TI - Clinical impact of correlative [123I]-FP-CIT brain imaging and neurological findings in suspect Parkinson's disease. AB - PURPOSE: Here we report our experience in a general hospital setting using [(123)I]-FP-CIT SPECT to diagnose patients with suspect Parkinson's disease (PD). MATERIALS AND METHODS: Thirty consecutive patients (19M, 11W, mean age: 61+/-13 years) were prospectively studied. Patients underwent MRI (27) at 1.5T or CT (3) when MRI was contraindicated, to rule out focal brain abnormalities. Motor and cognitive function were evaluated by neurologists with UPDRS and Hoehn e Yahr Scale. [(123)I]-FP-CIT striatal uptake, assessed with SPECT, was classified as normal, non-diagnostic, abnormal (unilateral or bilateral). Imaging results (SPECT+MRI) were correlated with the neurological findings. RESULTS: In 5 patients the [(123)I]-FP-CIT brain SPECT was normal, suggesting that their symptoms could be related to a benign disorder such as essential tremor. Two patients had non-diagnostic [(123)I]-FP-CIT brain SPECT, with MRI/CT findings compatible with subcortical cerebrovascular disease. In the remaining 23 patients abnormal striatal [(123)I]-FP-CIT uptake correlated with neurological findings, significantly increasing the probability of Parkinson's disease. In these patients MRI/CT scans were normal, or showed a mild BA, or mild cerebral vascular disease (mild CVD). CONCLUSIONS: Our results suggest that [(123)I]-FP-CIT scan could be used routinely in clinical practice to support the diagnosis of PD and to differentiate between other conditions. Moreover, FP-CIT could significantly impact treatment selection and follow-up of these patients. PMID- 15525896 TI - Comparison between indicators for the measurement of radiology activity volumes (Italy-USA). AB - PURPOSE: Different evaluation systems and indicators have recently been used to measure the activity volumes of Italian hospital departments, and in particular of Diagnostic Imaging Units. These measurements have mostly been based on more or less complex and repeatable indicators such as total accesses, accesses per imaging modality, type and number of exams. The aim of this study was to compare four models for measuring and evaluating productivity to assess their features and propose a common method for measuring activity volumes in a Diagnostic Imaging Unit. The models considered are: a) the numerical count, b) the model proposed by SNR-SAGO-SIRM, c) the model based on transfer prices in use in the Emilia Romagna Region (RER), d) the model used by the U.S. Health Care Financing Agency (HCFA-USA), based on a complex system of weights named RVUs (Relative Value Units). MATERIALS AND METHODS: The period under review considers two years of activity (2000-2001) at our Diagnostic Imaging Unit. The data were collected by grouping the radiological procedures into homogeneous groups (macroaggregates) which were then assessed with the four models. The reference parameters considered in order to produce homogeneous data were: the number of procedures per physician hour, the score per hour according to the SNR-SAGO-SIRM model, the score per hour according to the RER model, the number of work-RVUs per hour worked. With regard to the HCFA-USA system, the following indicators were used: the work component (work-RVU), the insurance component (malpractice RVU) and the technical component (practice expense-RVU), the equivalent units of physician time (FTE: Full Time Equivalent), such as the number of procedures per FTE, the difficulty index, and the number of RVUs per FTE. RESULTS: a) The total number of procedures was 55,884, the number of procedures per hour ranged from 2.43 (August 2000) to 4.20 (March 2000); based on the numerical count conventional radiology accounted for the most of the Unit's activity (40%). b) The total score according to the SNR-SAGO-SIRM model was 147,358; the weight of each physician hour ranged from 6.37 (August 2000) to 9.80 (October 2001). The SNR-SAGO-SIRM model indicates that the most significant macroaggregate in the Unit's activity was ultrasound (42%). c) The total score according to the RER model was 4,313,047, the weight of each physician hour varied between 159 (August 2000) and 316 (April 2000). Based on the RER model, CT (42%) accounted for most of the Unit's activity. d) According to the RVU model, the total number of work-RVUs was 37,619, and the physician weight per hour ranged from 1.45 (August 2000) to 2.86 (March 2000). The predominant method was ultrasound (35%); the number of total practice expense RVUs was 192,749; the month with the highest score was March 2000 (9,398), while the one with the lowest score was August 2000 (4,710); the total number of malpractice RVUs was 9,940, and the months with the highest scores were April 2000 (487) and March 2000 (487), while the month with the lowest score was August 2000 (243), and the modality carrying the highest insurance risks was MRI (38%). We also calculated the number of procedures per FTE (6,141), the number of work RVUs per FTE (4,134); the difficulty index resulting from the ratio between work RVUs and number of procedures (0.67); the number of work-RVUs per hour worked (3.06). CONCLUSIONS: Based on the numerical count, conventional radiology and ultrasound play a predominant role (40% and 34%, respectively, total 74%). This approach therefore fails to reflect the weight of more technologically advanced procedures. The SNR-SAGO-SIRM model gives adequate importance to the combination ''number- weight of patients'' among the macroaggregates analysed. The RER model rewards the use of more expensive technologies, as it assesses the overall weight of the service and not only the weight of the radiologist's activity. The RVU model, with its distribution of weights, differentiates the different work, cost, and insurance components of the macroaggregates. It also introduces an important aspect that is new to our professional and scientific culture: evaluation of the ''insurance component'', whose role will become increasingly important in Italy. The difficulty index (work-RVUs/no. of procedures), which expresses the ratio between the number of modalities and their complexity, is particularly interesting. This index, adjusted to reflect the Italian situation, might help to assess the true technological and scientific content of the department's activity. PMID- 15525897 TI - Influenza and pneumococcal vaccination coverage among persons aged > or =65 years and persons aged 18-64 years with diabetes or asthma--United States, 2003. AB - Vaccination of persons at risk for complications from influenza and pneumococcal disease is a key public health strategy for preventing associated morbidity and mortality in the United States. Risk factors include older age and medical conditions that increase the risk for complications from infections. During the 1990-1999 influenza seasons, more than 32,000 deaths each year among persons aged > or =65 years were attributed to complications from influenza infection. National health objectives for 2010 call for 90% influenza and pneumococcal vaccination coverage among noninstitutionalized persons aged > or =65 years and 60% coverage among noninstitutionalized persons aged 18-64 years who have risk factors (e.g., diabetes or asthma) for complications from infections. To estimate influenza and pneumococcal vaccination coverage among these populations, CDC analyzed data from the 2003 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report summarizes the results of that analysis, which indicated that 1) influenza vaccination levels among adults aged 18-64 with diabetes or asthma, 2) pneumococcal vaccination levels among adults aged 18-64 years with diabetes, and 3) influenza and pneumococcal vaccination levels among adults aged > or =65 years all were below levels targeted in the national health objectives for 2010. Moreover, vaccination coverage levels varied among states for both vaccines and both age groups. Innovative approaches and adequate, reliable supplies of vaccine are needed to increase vaccination coverage, particularly among adults with high risk conditions. PMID- 15525898 TI - Influenza vaccination and self-reported reasons for not receiving influenza vaccination among Medicare beneficiaries aged > or =65 years--United States, 1991 2002. AB - Annual influenza vaccination of the U.S. elderly population has been demonstrated as safe and effective in reducing the risks of illness, hospitalization, and death. The Medicare Current Beneficiary Survey (MCBS) has measured annual influenza vaccination rates since 1991; the latest data available are for the 2001-02 influenza season. Since 1996, self-reported reasons for not receiving influenza vaccine also have been measured. During 1991-2002, MCBS indicated a steady upward trend in vaccination coverage among Medicare beneficiaries, with the exception of the 2000-01 influenza season, when vaccine distribution was delayed. The most frequently cited reasons for not receiving influenza vaccine were 1) not knowing that influenza vaccination was needed and 2) concerns that vaccination might cause influenza or side effects. During the 2000-01 influenza season, vaccine shortage or unavailability was noted for the first time as an important reason for nonvaccination. Further efforts are needed to educate the elderly regarding the benefits of influenza vaccination and to address any concerns regarding the safety of the vaccine. PMID- 15525899 TI - Acute hepatitis B among children and adolescents--United States, 1990-2002. AB - Since the 1991 adoption of a comprehensive strategy to eliminate hepatitis B virus (HBV) transmission in the United States, the incidence of acute hepatitis B cases has declined steadily. Declines have been greatest among children born after the 1991 recommendations for universal infant hepatitis B vaccination were implemented. In 1995, the elimination strategy was expanded to include routine vaccination of all adolescents aged 11-12 years and, in 1999, to include children aged < or =18 years who had not been vaccinated previously. To describe the epidemiology of acute hepatitis B in children and adolescents in the United States, CDC analyzed notifiable disease surveillance data collected during 1990 2002 and data collected during 2001-2002 through enhanced surveillance of reported cases of acute hepatitis B in children born after 1990. This report summarizes the results of that analysis, which indicated that the rate of acute hepatitis B in children and adolescents decreased 89% during 1990-2002 and that racial disparities in hepatitis B incidence have narrowed. Many confirmed cases in persons born after 1990 occurred among international adoptees and other children born outside the United States. Continued implementation of the hepatitis B elimination strategy and accurate surveillance data to monitor the impact of vaccination are necessary to sustain the decline of acute hepatitis B among children. PMID- 15525900 TI - Blood mercury levels in young children and childbearing-aged women--United States, 1999-2002. AB - Exposure to high levels of mercury (Hg) can cause neurologic and kidney disorders. Because methylated Hg (methyl-Hg) in the aquatic environment accumulates in animal tissues up the food chain, persons in the United States can be exposed by eating freshwater fish, seafood, and shellfish. Exposure of childbearing-aged women is of particular concern because of the potential adverse neurologic effects of Hg in fetuses. To determine levels of total blood Hg in childbearing-aged women and in children aged 1-5 years in the United States, CDC's National Health and Nutrition Examination Survey (NHANES) began measuring blood Hg levels in these populations in 1999. This report summarizes NHANES results for 1999-2002 and updates previously published information. The findings confirmed that blood Hg levels in young children and women of childbearing age usually are below levels of concern. However, approximately 6% of childbearing aged women had levels at or above a reference dose, an estimated level assumed to be without appreciable harm (> or =5.8 microg/L). Women who are pregnant or who intend to become pregnant should follow federal and state advisories on consumption of fish. PMID- 15525901 TI - Outbreak of histoplasmosis among industrial plant workers--Nebraska, 2004. AB - In February 2004, the Nebraska Health and Human Services System (NHHSS) notified CDC about an outbreak of histoplasmosis among workers at a local agricultural processing plant (plant A). Three workers at the plant had acute, febrile, respiratory illness; two had serologic evidence of histoplasmosis. NHHSS and CDC conducted an investigation to determine the source of transmission and the extent of the outbreak. This report summarizes the findings of that investigation, which confirmed occupationally acquired histoplasmosis. Additional measures might be necessary to minimize risk for histoplasmosis among persons who work in the agricultural industry in areas where it is endemic. PMID- 15525902 TI - West Nile virus activity--United States, October 27-November 2, 2004. AB - During October 27-November 2, a total of 10 cases of human West Nile virus (WNV) illness were reported from eight states (Arizona, Georgia, Iowa, Michigan, New Mexico, Ohio, Oklahoma, and Pennsylvania). PMID- 15525903 TI - Muscarinic antagonist control of myopia: a molecular search for the M1 receptor in chick. AB - PURPOSE: Pirenzepine, an M1 selective muscarinic antagonist, is effective in slowing the progression of myopia in both humans and experimental animals, including chick. As an M1 selective antagonist, pirenzepine is considered to mediate its effect through M1 receptors. However, there is currently no report of the M1 receptor in chicken. Therefore, if the mechanism of action of pirenzepine is similar across species, either the drug mediates its effect through a non-M1 mechanism, or M1 muscarinic receptors are present in chicken. The aim of the present study was to determine whether a genetic template for the M1 receptor was expressed, or even present, in chick. METHODS: Polymerase Chain Reaction (PCR), and Southern and northern blotting analyses were used to search for M1 mRNA in chick ocular and brain tissues. PCR and Southern analyses were then used for searching the chick M1 gene and promoter. Appropriate rat positive controls were included throughout the study. RESULTS: Direct mRNA detection by northern analysis showed no evidence of M1 mRNA expression in the chick ocular and brain tissues studied. Identical results were obtained from PCR amplification and were further confirmed by Southern analysis. Similarly, no M1 gene or promoter sequences were detected by PCR or Southern analyses. Our methods were validated in every case by a positive finding in equivalent rat tissue and by detection of M2 and M4 mRNA expression in chick retina. CONCLUSIONS: Findings in this study suggest that the chick does not possess an M1 receptor. This finding is of primary interest to vision researchers in that it suggests pirenzepine is unlikely to mediate its inhibitory effect on the progression of myopia through an M1 receptor in chick. Alternative mechanisms of action are discussed. PMID- 15525906 TI - Thyroid nodules. PMID- 15525904 TI - Apolipoprotein E promoter polymorphisms do not have a major influence on the risk of developing primary open angle glaucoma. AB - PURPOSE: Primary open angle glaucoma (POAG) is a major cause of late onset visual failure of unknown etiology. Recent genetic association studies have implicated the apolipoprotein E (APOE) gene in the pathophysiology of primary open angle glaucoma, but there have been conflicting findings. METHODS: To resolve this issue we studied 140 cases and 73 controls that were carefully phenotyped, and used a logistic regression model to simultaneously analyze the effect of apolipoprotein E genotype and functional polymorphisms in the apolipoprotein E gene promoter while controlling for potentially confounding variables. RESULTS: We found no evidence of an association between the apolipoprotein E promoter region polymorphisms and primary open angle glaucoma. CONCLUSIONS: Apolipoprotein E promoter polymorphisms are unlikely to have a major impact on the pathophysiology of primary open angle glaucoma. PMID- 15525911 TI - Infection control practices for SARS in Lao People's Democratic Republic, Taiwan, and Thailand: experience from mobile SARS containment teams, 2003. AB - BACKGROUND: Despite available recommendations on infection control for severe acute respiratory syndrome (SARS), information is limited on actual practices in Asian hospitals during the epidemic. We describe practices observed by mobile SARS containment teams (mobile teams) during outbreak investigations. METHODS: We retrospectively summarized infection control practices observed in hospitals visited by mobile teams in the Lao People's Democratic Republic (PDR), Taiwan, and Thailand, during March and April 2003. RESULTS: Mobile teams investigated 22 reports of SARS in 20 hospitals (1, 5, and 14 hospitals in Lao PDR, Taiwan, and Thailand, respectively). Facilities ranged from urban hospitals with negative pressure isolation rooms and high-efficiency particulate air filtration to rural hospitals with patient rooms open to outside air circulation and intermittent running water. At the time of mobile team visits, 5 (25%) hospitals implemented infection control practices consistent with World Health Organization recommendations on visitor policies, private negative-pressure rooms, and personal protective equipment. CONCLUSIONS: Early in the SARS epidemic, mobile teams found wide variations in infection control practices and resources among Asian hospitals evaluating patients for SARS, indicating the importance of ongoing assessment during SARS preparedness. Mobile teams are one mechanism to assess practices and promote implementation of recommended infection control measures. PMID- 15525912 TI - Should self-assessment methods be used to measure compliance with handwashing recommendations? A study carried out in a French university hospital. AB - BACKGROUND: Implementation of a hand hygiene promotion program in a large university hospital required that we find a suitable method to assess health care workers' (HCWs) hand hygiene practices. This study aims at comparing direct observation and self-assessment methods. METHODS: Hand hygiene practices of 206 HCWs (physicians, nurses, and nurse assistants) in 25 care units were directly observed by trained auditors for 1 day. A week later, 1050 HCWs filled out a self assessment questionnaire on their compliance with handwashing indications (participation rate was 83%). RESULTS: Average global self-reported compliance rate (SRR) after patient care was similar to the observed rate (OBR) (74%). According to the type of care, differences between SRR and OBR were nonsignificant, except for change of infusion bag by nurses and nursing care by nurse assistants. Physicians and nurse assistants tended systematically to over evaluate their compliance, whereas nurses tended to under evaluate their compliance with hand hygiene recommendations. CONCLUSIONS: Mean compliance rates were higher than those reported in the literature but varied as a function of patient care activity and occupation of the HCW. A reinforced in-service educational program will be implemented that will target especially physicians and medical students. Self-assessment method, easy to use and inexpensive, gave encouraging results. The development of a broad-based, routine, self-assessment program is underway at Nantes University Hospital, but, before such a program can be implemented, reproducibility of these self-assessment indicators must be further confirmed. PMID- 15525913 TI - Field- and laboratory-based active dengue surveillance in Chennai, Tamil Nadu, India: observations before and during the 2001 dengue epidemic. AB - BACKGROUND: Dengue cases are reported every year in the city of Chennai, Tamil Nadu, India. Since April 2001, longitudinal field- and laboratory-based active dengue surveillance has been carried out in Chennai to study dengue trends. METHOD: A serologic survey of people in Chennai using the hemagglutination inhibition test (HIT) was performed to determine evidence of prior exposure to dengue virus infections. Dengue virus infections and their serotypes were demonstrated in vectors. The serum samples from clinical dengue patients were analyzed for dengue virus-specific immunoglobulins M (IgM) and G (IgG) antibodies by 2 commercial ELISA kits. RESULTS: There was an increase in the percentage of children with monotypic antibody responses to dengue in the later survey (April 2.2%, September 9.93%). DEN-3 serotype infections were demonstrated in male Aedes aegypti mosquitoes collected in September 2001. Dengue virus infection was diagnosed in 74.5% (143/192) of cases. While dengue-specific IgM responses were predominant among infants with dengue fever, IgG and mixed responses (M + G) were seen in 85% of the children with severe forms of dengue. CONCLUSION: The findings from these investigations suggest that antibody surveys in children and virus detection in vectors may be included as early warning system parameters in laboratory-based proactive dengue surveillance. PMID- 15525914 TI - Risk factors for surgical site infections following open-heart surgery in a Canadian pediatric population. AB - BACKGROUND: The rate and risk factors for surgical site infections (SSIs) in pediatric cardiac patients have not been well delineated. METHODS: All patients aged <18 years who had open-heart surgery at the Stollery Children's Hospital in the 1998-2002 period were followed. A case-control study was performed to examine risk factors for SSI. Controls were matched to cases according to National Nosocomial Surveillance System risk scores, age, and year of surgery. RESULTS: SSI incidence was 3.4% (0.9% superficial wound infections, .1% deep incisional surgical site infections, and 2.4% organ space surgical site infections). In the case-control study, the only risk factor that was statistically significant was the duration of surgery. There was a trend toward an increased incidence of SSI (P < .25) for children with failure to thrive, or for those who required inotropes or had an elevated serum lactate in the first 24 hours postoperation. CONCLUSION: In pediatric cardiac surgery, the risk of SSI increases with the duration of surgery. There is a need for prospective studies of potentially modifiable risk factors. PMID- 15525915 TI - Staphylococcus aureus and wounds: a review of tea tree oil as a promising antimicrobial. AB - Antibiotic-resistant bacteria continue to be a major health concern worldwide. In particular, Staphylococcus aureus, both methicillin-resistant and -sensitive, are of concern in their ability to cause difficult skin and underlying tissue infections. Melaleuca alternifolia oil (tea tree oil), an essential oil, has demonstrated promising efficacy in treating these infections. Tea tree oil has been used for centuries as a botanical medicine, and has only in recent decades surfaced in the scientific literature as a promising adjunctive wound treatment. Tea tree oil is antimicrobial, anti-inflammatory, and has demonstrated ability to activate monocytes. There are few apparent side effects to using tea tree oil topically in low concentrations, with contact dermatitis being the most common. Tea tree oil has been effective as an adjunctive therapy in treating osteomyelitis and infected chronic wounds in case studies and small clinical trials. There is a need for larger clinical trials to further examine efficacy of tea tree oil as an adjunctive wound therapy, as well as improved guidelines for developing plant-based medicines. PMID- 15525916 TI - Prevalence and characteristics of nosocomial infections in a Turkish university hospital. AB - BACKGROUND: The purpose of this study is to estimate the prevalence of nosocomial infections in a university hospital, as well as determining the groups at high risk of such infections. MATERIALS AND METHODS: Two surveys based on a modification of the British National Survey protocol for nosocomial infection were conducted in July and December 1998. RESULTS: In the first survey, hospital infections were found in 41 (13.4%) of the patients, and in the second survey in 34 (10.9%). The study showed that the risk of nosocomial infection was associated with being in the intensive care unit, undergoing surgery, and invasive procedures. CONCLUSION: Prevalence data are consistent with results reported in many other regions of the world. These findings provide the principal information for future surveillance in association with prevention programs in Turkish hospitals. PMID- 15525917 TI - Antimicrobial susceptibility of microorganisms recovered from intraabdominal infections at Belo Horizonte, Brazil. AB - We evaluated antimicrobial susceptibility patterns of microorganisms isolated from intraabdominal infection of Brazilian patients, by agar dilution, agar diffusion, and E test. Among the strictly anaerobes, 57.7% were resistant to penicillin, 28.2% to clindamycin, and 9.9% to metronidazole. The majority of Escherichia coli and Staphylococcus were sensitive and resistant to almost all drugs, respectively. Half of Candida samples were resistant to itraconazole. Our data reinforce the importance of this kind of study to support rational antimicrobial therapy. PMID- 15525918 TI - Universal SARS preventive measures in an obstetrics unit: experience of health care staff. AB - BACKGROUND: Severe acute respiratory syndrome (SARS) epidemics have affected populations in many countries, including Hong Kong. This disease is infectious, especially in hospital settings. Health care workers have expressed great concern, including those working in obstetrics wards, defined as high-risk areas. METHOD: Four weeks after implementation of universal precautionary measures at a teaching hospital in Hong Kong, a survey of the health care staff was conducted to identify their feelings and opinions. RESULTS: In spite of general knowledge about SARS epidemics and related mortality, most respondents stated that universal precautionary measures were not very necessary, especially in the obstetrics ward. In addition, respondents were generally dissatisfied with the measures, as most items imposed extra work, inconvenience, and burdens on the staff. CONCLUSION: Our findings reported the views and satisfaction levels of the front-line staff of an obstetric unit concerning precautionary measures against SARS. The importance of individualized design and implementation of infection control measures is highlighted and discussed. PMID- 15525919 TI - Patient safety tools to avoid disaster: Health care facility management of biologic agents, SARS, and uncommon contagion. PMID- 15525920 TI - Promoting quality through surveillance of surgical site infections: five prevention success stories. AB - BACKGROUND: Public reports of successful quality improvement efforts are useful, but seldom available. We present 5 successful efforts to prevent surgical site infections (SSIs) with the use of prospectively collected surveillance data. METHODS: Before-and-after intervention studies were conducted in 5 acute care public hospitals in the national surveillance network for SSI in The Netherlands from 1992 to 2000. Patients undergoing surgery for total hip prosthesis (3 hospitals), knee prosthesis (2 hospitals), prosthesis of the femur head (1 hospital), or appendectomy (1 hospital) were included. Included were 1066 patients before intervention, and 1269 patients after intervention. Multidisciplinary evaluation of infection control policy led to subsequent changes of infection control measures, mainly involving the discipline of staff and organization of perioperative infection prevention procedures. RESULTS: All 5 hospitals drastically reduced their SSI rates to the national average or below. Absolute declines ranged from 2.1% to 13.9%, but not all reductions were statistically significantly different from 0%. CONCLUSION: Surveillance results provide a basis for improvement of infection prevention. PMID- 15525921 TI - An unusual case of tophaceous gout involving the anterior cruciate ligament. AB - Abstract We describe a very unusual case of a 49-year-old man with tophaceous gout involving and infiltrating the base of the anterior cruciate ligament of the left knee. To our knowledge, such a case has never been reported in the literature, although gout is well known and widely studied. Magnetic resonance imaging findings and differential diagnosis were analyzed before arthroscopy. Tophaceous gout was suspected at arthroscopy and confirmed on pathologic examination. Treatment consisted of partial removal of the tumor by arthroscopy, conserving the anterior cruciate ligament. PMID- 15525922 TI - Enhancement of tendon graft osteointegration using mesenchymal stem cells in a rabbit model of anterior cruciate ligament reconstruction. AB - PURPOSE: To study the effect of coating tendon grafts with mesenchymal stem cells (MSCs) on the rate and quality of graft osteointegration in anterior cruciate ligament (ACL) reconstruction. TYPE OF STUDY: Animal model. METHODS: Bilateral ACL reconstructions using hamstring tendon autografts were performed on 48 adult rabbits. Grafts were coated with MSCs in a fibrin glue carrier in one limb, and fibrin glue only in the other. Assessment was done at 2, 4, and 8 weeks. Histologic analysis was carried out using standard and immunohistochemical stains. Biomechanical testing of force and stiffness during loading to ultimate failure was performed. RESULTS: Control reconstructions showed mature scar tissue with some Sharpey's-like fibers spanning the tendon-bone interface at 8 weeks. The MSC-enhanced reconstructions had large areas of cartilage cells at the tendon bone junction at 2 weeks. By 8 weeks, a mature zone of cartilage was seen gradually blending from bone into the tendon grafts. This zone stained strongly for type II collagen and showed histologic characteristics similar to normal rabbit ACL insertions. Biomechanically, there was no statistical difference between limbs at 2 and 4 weeks. At 8 weeks, the MSC-enhanced grafts had significantly higher failure load and stiffness. CONCLUSIONS: Coating of tendon grafts with MSCs results in healing by an intervening zone of cartilage resembling the chondral enthesis of normal ACL insertions rather than collagen fibers and scar tissue. MSC-enhanced ACL reconstructions perform significantly better than controls on biomechanical testing. CLINICAL RELEVANCE: Enhancement of tendon graft osteointegration with MSCs is a novel method offering the potential for more physiologic and biomechanically stronger ligament reconstructions. PMID- 15525923 TI - Histologic changes in articular cartilage after medial meniscus replacement in rabbits. AB - PURPOSE: Meniscal allografts show capsular ingrowth, but it remains to be established whether meniscal transplantation can prevent long-term degeneration of articular cartilage. This study examined whether immediate or delayed transplantation of the medial meniscus can protect the knee from degenerative changes. TYPE OF STUDY: Experimental study. METHODS: Thirty-five rabbits were divided into 5 groups. Three rabbits developed infective arthritis and were excluded from the study. Group A (6 animals) and group C (6 animals) had meniscectomy only; group B (7 animals) and group D (6 animals) underwent meniscal transplantation immediately after meniscectomy; group E (7 animals) had delayed transplantation 6 weeks after meniscectomy. Six nonoperated knees served as controls. Histologic changes of the articular cartilage were examined 6 weeks (groups A, B) and 1 year (groups C, D, E, controls) after surgery. RESULTS: All operated groups showed more histologic changes than the control group (P < .00001 for both the medial tibial plateau and medial femoral condyle in all groups). At 6 weeks follow-up, no differences were found between the postmeniscectomy group and the transplanted group. At 1 year, immediately transplanted knees showed less degenerative changes than meniscectomized knees (P < .0001 for medial tibial plateau and P < .005 for medial femoral condyle). Delayed transplantation resulted in more degenerative changes than both meniscectomy only and immediate transplantation (for both comparisons, P < .00001 for both medial tibial plateau and medial femoral condyle). CONCLUSIONS: Immediate meniscal transplantation in rabbits has a protecting effect on articular cartilage for the long-term whereas delayed transplantation leads to even more degenerative changes than meniscectomy only. CLINICAL RELEVANCE: If these results are applicable to humans, they would support immediate, not delayed, transplantation after removal of a meniscus. PMID- 15525924 TI - When to inject analgesic agents intra-articularly in anterior cruciate ligament reconstruction: before or after tourniquet releasing. AB - PURPOSE: To evaluate 2 different applications of an intra-articular (IA) mixture composed of 40 mL 0.25% bupivacaine and 2 mg morphine in anterior cruciate ligament (ACL) reconstructions, before and after tourniquet release. TYPE OF STUDY: Prospective, double-blind, randomized, clinical trial. METHODS: Forty patients who underwent ACL reconstruction were prospectively randomized into 2 groups. Group 1 was injected with IA morphine with bupivacaine 10 minutes before tourniquet release. Group 2 was injected with IA morphine with bupivacaine injection after tourniquet release, after a 30-minute period of waiting with closed drainage. Visual analog scale scores were recorded at 10 and 30 minutes and at 1, 2, 4, 8, 12, and 24 hours after surgery. Detailed records were kept of the amount of analgesic drug required. RESULTS: In pain scores, there was no difference between the 2 groups except for 30 minutes postoperatively (P = .005). The first analgesic requirement time was significantly lower in group 2 (P < .001) and the total amount of opioid used was lower in group 2 (P = .037) than those in group 1. There was no difference between the 2 groups in terms of operation time (P = .484), although there was a significant difference between groups in terms of tourniquet time (P = .024). CONCLUSIONS: We conclude that IA analgesic injection after tourniquet release is more beneficial to obtain postoperative analgesia in ACL reconstructions than the same injection before tourniquet release. LEVEL OF EVIDENCE: Level I. PMID- 15525925 TI - Graft-bone motion and tensile properties of hamstring and patellar tendon anterior cruciate ligament femoral graft fixation under cyclic loading. AB - PURPOSE: To assess longitudinal graft-bone motion and tensile properties of the femur-anterior cruciate ligament (ACL) graft fixation-ACL graft complex based on the hypothesis that there is little difference in graft-bone motion between suspensory and aperture hamstring ACL femoral graft fixation techniques, and between hamstring and patellar tendon ACL femoral graft fixation techniques. TYPE OF STUDY: In vitro biomechanical study using human cadavers. METHODS: The distal femur-ACL graft fixation-ACL graft complex was cyclically loaded between 50 and 250 N at 1 Hz for 1,000 cycles with the direction of the load applied parallel to the axis of the femoral bone tunnel. Graft-bone motion was measured indirectly using retroreflective markers and a video motion-analysis system. Tensile testing to failure was performed at 1 mm/sec for fixation techniques completing 1,000 cycles without fixation failure. RESULTS: Among the hamstring fixation techniques, 4 of 13 Bio-Interference screws (Arthrex, Naples, FL), 2 of 12 LinX HT fasteners (DePuy Mitek, Norwood, MA), and 1 of 11 TransFix cross-pins (Arthrex) failed before completing 1,000 cycles. Five of 13 patellar tendon grafts fixed with metal interference screws, and 2 of 12 patellar tendon grafts fixed with a plastic button and No. 5 sutures failed before completing 1,000 cycles. Suture/button fixation of patellar tendon grafts resulted in significantly more graft-bone motion than hamstring tendon grafts fixed using the Bone Mulch Screw (Arthrotek, Warsaw, IN), or interference screw fixation of patellar tendon and hamstring grafts. Otherwise, there was no significant difference in graft-bone motion among the various hamstring fixation techniques or the various hamstring fixation techniques and interference screw fixation of patellar tendon grafts. Maximum graft-bone displacement after cyclic loading was significantly greater for hamstring grafts fixed with the EndoButton and EndoButton Tape (Smith & Nephew Endoscopy, Andover, MA) compared with the other hamstring fixation techniques and interference screw fixation of patellar tendon grafts. All fixation techniques except hamstring tendon grafts fixed with the Bio Interference screw achieved at least 59% of maximum graft-bone displacement after 20 cycles. Hamstring tendon grafts fixed with the EndoButton CL were significantly stronger than all other hamstring and patellar tendon fixation methods. Patellar tendon grafts fixed with interference screws and hamstring tendon grafts fixed with interference screws and the Bone Mulch Screw and TransFix were significantly stiffer than hamstring tendon grafts fixed with the EndoButton CL. CONCLUSIONS: There is no significant difference in graft-bone motion between aperture and suspensory femoral fixation methods when the stiffness of the femur-ACL graft fixation-ACL graft complex is similar. CLINICAL RELEVANCE: The small differences in graft-bone motion reported in our study provide further evidence that graft-tunnel motion or the so-called bungee effect is unlikely to be the primary cause of radiographic bone tunnel enlargement following ACL reconstruction. PMID- 15525927 TI - A comparison of bone tunnel measurements made using computed tomography and digital plain radiography after anterior cruciate ligament reconstruction. AB - PURPOSE: The purpose of this study was to compare measurements of bone tunnel enlargement after anterior cruciate ligament (ACL) reconstruction using digital plain radiography and computed tomography (CT). TYPE OF STUDY: Within-subjects comparison of 2 imaging techniques. METHODS: Twenty-two patients who had undergone primary ACL reconstruction had anteroposterior and lateral digital plain radiographs and a CT scan of their operated knee 12 months after surgery. Using digital calipers, the sclerotic margins of each tunnel were measured at the widest point in each plane. Measurements were corrected for magnification and expressed as a percentage change in tunnel width compared with the size of the drill bit used at surgery. RESULTS: In all but 1 instance, bone tunnels seen on CT were also seen on the corresponding digital radiograph. In the single incidence in which a bone tunnel was identified on the CT scan but not on the radiograph, the tunnel had the smallest diameter of the group and had decreased since surgery by 30%. There was no difference between the group-averaged CT and radiographic measurements for the tibia, but femoral tunnel radiographic measurements were clearly larger than CT, particularly for the anteroposterior view (P < .001). Moderate discrepancy was noted between individual radiograph and CT measurements. CONCLUSIONS: Digital plain radiography appears to be satisfactory for detecting bone tunnel enlargement following ACL reconstruction. Thus it may offer a time- and cost-effective means for monitoring this phenomenon. LEVEL OF EVIDENCE: Level I. PMID- 15525926 TI - Arthroscopic all-inside suture repair of medial meniscus lesion in anterior cruciate ligament--deficient knees: results of second-look arthroscopies in 39 cases. AB - PURPOSE: To evaluate by second-look arthroscopy the clinical results of arthroscopic all-inside sutures with hook using 2 posteromedial portals for medial meniscus posterior horn (MMPH) tears in patients who underwent concurrent anterior cruciate ligament (ACL) reconstruction. TYPE OF STUDY: Case series. METHODS: From May 1997 to June 2001, 78 knees underwent surgery for MMPH tears with arthroscopic all-inside sutures with hook and concurrent ACL reconstruction. Among them, 39 patients were evaluated with follow-up second-look arthroscopy. All MMPH tears were repaired by arthroscopic all-inside suture technique using 2 posteromedial portals. The second-look arthroscopy was performed, on average, 19 months (range, 6 to 40 months) after the ACL reconstruction and meniscal repair. They were divided into complete healing, incomplete healing, and failure groups. Tear size, type, and location were analyzed. We determined clinical criteria for success in meniscal status as (1) not positive for 4 clinical objective parameters--joint line pain and tenderness, locking or catching, recurrent effusions, and McMurray test; and (2) complete healing of all-inside sutured meniscus during second-look arthroscopy. RESULTS: Among 39 knees assessed by second-look arthroscopy, 32 (82.1%) knees showed complete healing and 6 (15.4%) showed incomplete healing without any positive findings of the clinical symptoms. Furthermore, all cases in the incompletely healed group had complete healing of the posterior horn, which was sutured by an all-inside suture; the incomplete healing was observed between the junction areas of all-inside to inside-out sutures, which was mainly at the posteromedial corner. The success rate was 97.4% (38 of 39 patients) overall for all-inside suture healing. The 1 patient (2.6%) with clinical failure had a retear of the repaired site. The average knee scores improved and all showed less than 2-mm side-to-side difference on KT-2000 testing. CONCLUSIONS: Arthroscopic all-inside vertical suture using a suture hook resulted in a high rate of healing even in large and complex vertical tears. This suturing can be one of the optimal treatments for MMPH tears greater than 1 cm during concurrent ACL reconstruction. LEVEL OF EVIDENCE: Level IV, Therapeutic Study, Cases Series (no, or historical, control group). PMID- 15525928 TI - A new weight-bearing meniscal test and a comparison with McMurray's test and joint line tenderness. AB - PURPOSE: The purpose of this study was to describe a new weight-bearing McMurray's test (Ege's test) and to compare its diagnostic value with McMurray's test and joint line tenderness (JLT). We also aimed to determine if associated lesions had any effect on the diagnostic values of the 3 tests. TYPE OF STUDY: Prospective controlled trial, clinical study. METHODS: The study group consisted of 150 consecutive patients who had had symptoms related to intra-articular knee pathology, and arthroscopic diagnoses were used as the gold standard. RESULTS: There were a total of 211 diagnoses on arthroscopy. A meniscus tear was found in 127 of the 150 patients; 90 had medial, 28 had lateral, and 9 had tears of both menisci. There were no statistically significant differences between the 3 tests in detecting a meniscal tear ( P > .05). However, better accuracy, sensitivity, and specificity rates were obtained with Ege's test for medial meniscal lesions (71%, 67%, and 81%, respectively). JLT also gave superior accuracy rates (71%), but the specificity of Ege's test was apparently higher than JLT (81% v 44%). The highest positive predictive value was also obtained with Ege's test (86%), whereas a superior negative predictive value was obtained with JLT (67%) in medial meniscal tears. Lateral meniscal tears were diagnosed more accurately than medial meniscal tears, and Ege's test gave results superior to the others (84%, 64%, 90% for accuracy, sensitivity, and specificity, respectively). Higher positive predictive values were obtained with McMurray's and Ege's tests than with JLT, but similar negative predictive values were achieved in all. A torn anterior cruciate ligament did not decrease the diagnostic values of the 3 tests, whereas the number of associated lesions in the knee negatively affected the diagnostic capabilities of the tests. CONCLUSIONS: Accuracies of traditional clinical meniscus tests may be improved by including Ege's test in the clinical examination. LEVEL OF EVIDENCE: Level II, diagnostic. PMID- 15525929 TI - Chronic labral tears: value of magnetic resonance arthrography in evaluating the glenoid labrum and labral-bicipital complex. AB - PURPOSE: In the magnetic resonance (MR) evaluation of the glenoid labrum, all studies to our knowledge have included mixed populations of patients, those with acute shoulder injuries as well as patients with chronic shoulder disorders. The focus of this investigation was to assess the effectiveness of MR arthrography in patients with chronic labral tears, excluding those with acute injuries. TYPE OF STUDY: Prospective case series. METHODS: Conventional MR images and MR arthrograms were obtained in 36 patients from April 1994 to April 1997. A single experienced musculoskeletal radiologist read all MR images. Each patient subsequently underwent shoulder arthroscopy performed by a single highly experienced shoulder arthroscopist. Detailed arthroscopic reports were then reviewed and compared with the MR findings documented before surgery, with arthroscopic findings being the standard of reference for comparison. Inclusion criteria required greater than 6 months of shoulder symptoms before imaging, thus eliminating acute injuries. RESULTS: SLAP lesions were diagnosed at the time of surgery in 11 of 36 patients (31%). The sensitivity was 100% (11 of 11 patients) and the specificity was 88% (22 of 25 patients). Accuracy for SLAP lesions was 92% (33 of 36 patients). Anterior labral tears were diagnosed surgically in 12 of 36 patients (33%). The sensitivity was 86% (12 of 14 patients) and specificity was 86% (19 of 22 patients). Accuracy for labral tears was 86% (31 of 36 patients). CONCLUSIONS: MR arthrography is an accurate technique for assessing the glenoid labrum in patients with chronic labral tears. LEVEL OF EVIDENCE: Level I. PMID- 15525931 TI - Anterior cruciate ligament reconstruction: which graft is best? AB - Abstract For the last 4 decades, since the initial use of the patellar tendon for anterior cruciate ligament (ACL) reconstruction, there has been controversy regarding the ideal graft choice for this procedure. Beside bone-patellar tendon bone autografts, several other graft choices have become popular, including hamstring tendon and a variety of allografts. Within the past 5 years, several randomized and nonrandomized studies have compared the graft choices in ACL reconstruction. However, the question still remains: Is there an ideal graft for ACL reconstruction? The purpose of this review is to assess the most recent data, identifying if there truly is an ideal graft choice. PMID- 15525930 TI - Arthroscopic bicipital sheath repair: two-year follow-up with pulley lesions. AB - PURPOSE: The purpose of this study was to evaluate arthroscopic repair in patients who had lesions of both the subscapularis insertion/medial head of the coracohumeral ligament and the lateral head of the coracohumeral ligament and supraspinatus tendon (a type 5 biceps subluxation/instability classification), and to determine if primary repair of the torn structures used to reconstruct the bicipital sheath was associated with a high biceps rupture rate. The null hypothesis, that there is no difference between preoperative and postoperative outcomes, was tested. TYPE OF STUDY: Prospective cohort. METHODS: Since 1995, the author has had 18 patients who had lesions that affected both the medial and lateral wall of the bicipital sheath. An adjunct was added if tendonitis was present with fraying, and the biceps tendon was debrided if the fraying consisted of 50% or less the width of the tendon. This was chosen arbitrarily. Greater than 50% fraying of the biceps tendon was treated with repair of the supraspinatus and subscapularis. The biceps tendon was treated with tenotomy or tenodesis in these cases and these patients were not included in this study. This article reports on the repair technique and results having a minimum of 2-year follow-up. RESULTS: There were 12 male patients (age range, 45 to 80 years; average, 62 years) and 6 female patients (age range, 50 to 85 years; average, 66 years). The dominant extremity was involved in 12 of the 16 extremities. Preoperative, ASES Index, Total Constant scores, Subjective Constant scores, Objective Constant scores, visual analog pain scales, and percent function were 31 +/- 19, 53 +/- 13, 12 +/- 8, 41 +/- 8, 7 +/- 3, and 42 +/- 17, respectively. Postoperative scores were 80 +/- 14, 77 +/- 10, 30 +/- 4, 47 +/- 7, 2 +/- 2, and 84 +/- 14, respectively. The null hypothesis was rejected at a level of P = .001, .001, .001, .05, .001, and .001, respectively. CONCLUSIONS: There was 1 biceps disruption in this cohort following repair, for an incidence rate of 6%. There were 2 patients, active tennis players, who had recurrence of biceps inflammation in the follow-up period with no evidence of biceps subluxation. The arthroscopic technique reported is a primary repair used to reconstruct the normal structures of the groove. This may explain why previous recommendations not to reconstruct the groove because of the high biceps disruption rate have been noted previously. This study did not deepen the groove, tubulize the biceps tendon, or close the rotator interval in nonanatomic fashion. This arthroscopic technique is technically feasible and can alleviate the symptoms of biceps tendon inflammation and/or subluxation in the majority of cases in this cohort. LEVEL OF EVIDENCE: Level IV, Case Series. PMID- 15525932 TI - Surgical fixation of massive osteochondritis dissecans lesion using cylindrical osteochondral plugs. AB - Abstract The prognosis of osteochondritis dissecans (OCD) depends on the age of manifestation and the size of the lesion. Before epiphyseal closure, OCD usually responds well to the restriction of the patient's sporting activities. Surgical intervention is not always necessary. However, if the disease manifests after epiphyseal closure, it often needs surgical treatment because the lesion is usually found at an advanced stage (International Cartilage Repair Society [ICR] OCD II or more). Furthermore, if a massive lesion develops to separation (ICR OCD II or III) or displaced loose bodies (ICR OCD IV), despite restriction of sporting activities, the prognosis is poor. This report discusses 2 cases of OCD. One patient had a massive stage ICR I lesion, although the disease was found before epiphyseal closure. In the other patient, a 22-year-old man, a large separated fragment (ICR OCD IV) was found after epiphyseal closure. The separated surfaces of the fragment and the mother bed had become sclerotic. Cylindrical osteochondral grafts using jigs for Mosaicplasty (Smith & Nephew, Andover, MA) were performed on the 2 patients, both of whom made successful recoveries. These cases suggest that cylindrical osteochondral grafting is a useful procedure for difficult cases of large lesion size or late manifestation. PMID- 15525933 TI - The complete type of suprapatellar plica in a professional baseball pitcher: consideration of a cause of anterior knee pain. AB - Abstract We report the case of a professional baseball pitcher who achieved complete relief of an anterior knee pain after resection of a complete type of suprapatellar plica under arthroscopy. The 27-year-old male professional baseball pitcher had complained of right anterior knee pain while pitching for more than 2 years. On physical examination, the mobility of his patella was limited and he complained of tenderness along the joint line of his lateral patellofemoral joint. The preoperative patellofemoral axial radiograph showed osteophyte formation on the lateral edge of the patella. On preoperative magnetic resonance imaging, a septum dividing the suprapatellar pouch was found. Arthroscopy revealed obvious cartilaginous damage on the lateral facet of the patella and facing trochlea of the femoral condyle. There was a complete type of suprapatellar plica. The suprapatellar plica seemed to be anchoring the patella, which reduced the mobility of the patella. After resecting the complete type of suprapatellar plica under arthroscopy, the mobility of the patella was improved. Because his anterior knee pain while pitching reduced rapidly postoperatively, he could return to regular baseball. We consider that the main cause of his anterior knee pain was cartilaginous damage on the patella and the trochlea, which might be accelerated by the existence of the suprapatellar plica that reduced the mobility of the patella. PMID- 15525934 TI - Tendon-to-bone healing of a quadrupled hamstring tendon graft fixed with biodegradable screws in an immature athlete: a radiologic, arthroscopic, histologic, and electromicroscopic investigation. AB - Abstract Quadrupled hamstring tendon is one of the most commonly used ligamentous substitutes in anterior cruciate ligament reconstruction. We describe a case of a young athlete who sustained a rerupture of a semitendinosus-gracilis autograft fixed with biodegradable screws 2 years after primary surgery. Radiologic, arthroscopic, histologic, and electromicroscopic investigation showed tendon-to bone healing of the soft tissue graft and partial degradation of the interference screws. PMID- 15525935 TI - Lateral patellofemoral ligament reconstruction. AB - Abstract Medial dislocation of the patella is a disabling condition; there are several reports in the literature describing this condition and its association with failed lateral retinacular release. The diagnosis and treatment of medial subluxation of the patella may be difficult. Direct repair or imbrication of the lateral retinaculum provides initial stability but a noticeable increase in medial excursion usually reappears. In this article, we describe a simple and reproducible technique to reconstruct the lateral patellofemoral ligament with autogenous tissue that is based on the basic principles of all ligament reconstruction. Reconstruction of the lateral patellofemoral ligament is useful in eliminating the symptoms related to medial instability of the patella after failed lateral retinacular release; however, it must be considered a salvage procedure because it does not address the pathomechanics that led to the initial patellofemoral symptoms. PMID- 15525936 TI - Arthroscopically assisted core decompression of the proximal humerus for avascular necrosis. AB - Abstract Core decompression has been described as an effective treatment for early stage avascular necrosis of the humeral head. This article describes the technique for arthroscopically assisted core decompression of the humeral head using a transtibial anterior cruciate ligament guide. This technique provides an advantage over the open procedure by avoiding soft-tissue stripping or damaging the biceps tendon or ascending branch of the anterior humeral circumflex artery, as well as providing accurate and safe placement of the core biopsy tract and the ability to address other intra-articular pathology. PMID- 15525937 TI - Arthroscopic supraspinatus tendon repair using suture anchors and a modified Mason-Allen technique: an intra-articular approach. AB - Abstract Arthroscopic repair of supraspinatus tendon tears has had clinical results comparable to those of open procedures. The arthroscopic subacromial approach is well described in the literature but is still limited to skilled arthroscopic surgeons owing to the technical difficulties of the method. We present an intra-articular approach for supraspinatus tendon repair using suture anchors and a modified Mason-Allen technique. A major advantage of this technique is an excellent visualization of the procedure that optimizes the preparation of the lesion, positioning of the suture anchors, and, especially, grasping of the tendon with suture devices. The easy and exact positioning of the sutures allows creation of a modified arthroscopic Mason-Allen stitch. The result is a very stable initial tendon-to-bone fixation, which is necessary for a sufficient osteofibroblastic integration. PMID- 15525938 TI - Regulation of p53 activity through lysine methylation. AB - p53 is a tumour suppressor that regulates the cellular response to genotoxic stresses. p53 is a short-lived protein and its activity is regulated mostly by stabilization via different post-translational modifications. Here we report a novel mechanism of p53 regulation through lysine methylation by Set9 methyltransferase. Set9 specifically methylates p53 at one residue within the carboxyl-terminus regulatory region. Methylated p53 is restricted to the nucleus and the modification positively affects its stability. Set9 regulates the expression of p53 target genes in a manner dependent on the p53-methylation site. The crystal structure of a ternary complex of Set9 with a p53 peptide and the cofactor product S-adenosyl-l-homocysteine (AdoHcy) provides the molecular basis for recognition of p53 by this lysine methyltransferase. PMID- 15525939 TI - Methylated lysine 79 of histone H3 targets 53BP1 to DNA double-strand breaks. AB - The mechanisms by which eukaryotic cells sense DNA double-strand breaks (DSBs) in order to initiate checkpoint responses are poorly understood. 53BP1 is a conserved checkpoint protein with properties of a DNA DSB sensor. Here, we solved the structure of the domain of 53BP1 that recruits it to sites of DSBs. This domain consists of two tandem tudor folds with a deep pocket at their interface formed by residues conserved in the budding yeast Rad9 and fission yeast Rhp9/Crb2 orthologues. In vitro, the 53BP1 tandem tudor domain bound histone H3 methylated on Lys 79 using residues that form the walls of the pocket; these residues were also required for recruitment of 53BP1 to DSBs. Suppression of DOT1L, the enzyme that methylates Lys 79 of histone H3, also inhibited recruitment of 53BP1 to DSBs. Because methylation of histone H3 Lys 79 was unaltered in response to DNA damage, we propose that 53BP1 senses DSBs indirectly through changes in higher-order chromatin structure that expose the 53BP1 binding site. PMID- 15525941 TI - Bad funding. PMID- 15525940 TI - The role of autophagy during the early neonatal starvation period. AB - At birth the trans-placental nutrient supply is suddenly interrupted, and neonates face severe starvation until supply can be restored through milk nutrients. Here, we show that neonates adapt to this adverse circumstance by inducing autophagy. Autophagy is the primary means for the degradation of cytoplasmic constituents within lysosomes. The level of autophagy in mice remains low during embryogenesis; however, autophagy is immediately upregulated in various tissues after birth and is maintained at high levels for 3-12 h before returning to basal levels within 1-2 days. Mice deficient for Atg5, which is essential for autophagosome formation, appear almost normal at birth but die within 1 day of delivery. The survival time of starved Atg5-deficient neonates (approximately 12 h) is much shorter than that of wild-type mice (approximately 21 h) but can be prolonged by forced milk feeding. Atg5-deficient neonates exhibit reduced amino acid concentrations in plasma and tissues, and display signs of energy depletion. These results suggest that the production of amino acids by autophagic degradation of 'self' proteins, which allows for the maintenance of energy homeostasis, is important for survival during neonatal starvation. PMID- 15525942 TI - Good eggs. PMID- 15525943 TI - China takes steps to secure pole position in primate research. PMID- 15525944 TI - Early embryos fuel hopes for shortcut to stem-cell creation. PMID- 15525945 TI - Climate change clouds commercial licence to krill. PMID- 15525946 TI - Green groups baulk at joining nanotechnology talks. PMID- 15525947 TI - Mouse sequencing plan aims to boost models. PMID- 15525948 TI - Standing room only signals US zeal for Earth imaging. PMID- 15525950 TI - British Museum bids to stop illicit traders using eBay. PMID- 15525949 TI - EPA accused of conflict of interest over chemicals study. PMID- 15525951 TI - Beta-blocker goes on trial as asthma therapy. PMID- 15525954 TI - Medicinal leeches: stuck on you. PMID- 15525955 TI - Marine conservation: Sink or swim. PMID- 15525956 TI - Industry funding doesn't influence our reports. PMID- 15525957 TI - US rules on tech transfer to foreign nationals. PMID- 15525958 TI - European council should back young investigators. PMID- 15525964 TI - Making sense. PMID- 15525965 TI - Wandering nostrils. PMID- 15525967 TI - Cell biology: adhesion articulated. PMID- 15525968 TI - Land management: forests, fires and climate. PMID- 15525969 TI - Cardiovascular biology: how genes know their place. PMID- 15525970 TI - Obituary: Fred Lawrence Whipple (1906-2004). PMID- 15525972 TI - Archaeology: formulation of a Roman cosmetic. AB - The discovery of a small tin canister in London during archaeological excavations of a Roman temple precinct, dated to the middle of the second century AD, is a landmark in the study of this class of artefact. Such discoveries from the Roman world are rare and this is the only one to be found so far with its lid and contents--a whitish medicinal or cosmetic cream--providing a unique opportunity for us to study the ancient formulation. PMID- 15525973 TI - Biophysics: water-repellent legs of water striders. AB - Water striders (Gerris remigis) have remarkable non-wetting legs that enable them to stand effortlessly and move quickly on water, a feature believed to be due to a surface-tension effect caused by secreted wax. We show here, however, that it is the special hierarchical structure of the legs, which are covered by large numbers of oriented tiny hairs (microsetae) with fine nanogrooves, that is more important in inducing this water resistance. PMID- 15525975 TI - The fertility riddle. PMID- 15525976 TI - Age is no barrier... PMID- 15525977 TI - Waiting for the second coming. PMID- 15525979 TI - Seeds of concern. PMID- 15525980 TI - Resourceful imprinting. PMID- 15525981 TI - Insights into assembly from structural analysis of bacteriophage PRD1. AB - The structure of the membrane-containing bacteriophage PRD1 has been determined by X-ray crystallography at about 4 A resolution. Here we describe the structure and location of proteins P3, P16, P30 and P31. Different structural proteins seem to have specialist roles in controlling virus assembly. The linearly extended P30 appears to nucleate the formation of the icosahedral facets (composed of trimers of the major capsid protein, P3) and acts as a molecular tape-measure, defining the size of the virus and cementing the facets together. Pentamers of P31 form the vertex base, interlocking with subunits of P3 and interacting with the membrane protein P16. The architectural similarities with adenovirus and one of the largest known virus particles PBCV-1 support the notion that the mechanism of assembly of PRD1 is scaleable and applies across the major viral lineage formed by these viruses. PMID- 15525983 TI - An interplanetary shock traced by planetary auroral storms from the Sun to Saturn. AB - A relationship between solar activity and aurorae on Earth was postulated long before space probes directly detected plasma propagating outwards from the Sun. Violent solar eruption events trigger interplanetary shocks that compress Earth's magnetosphere, leading to increased energetic particle precipitation into the ionosphere and subsequent auroral storms. Monitoring shocks is now part of the 'Space Weather' forecast programme aimed at predicting solar-activity-related environmental hazards. The outer planets also experience aurorae, and here we report the discovery of a strong transient polar emission on Saturn, tentatively attributed to the passage of an interplanetary shock--and ultimately to a series of solar coronal mass ejection (CME) events. We could trace the shock-triggered events from Earth, where auroral storms were recorded, to Jupiter, where the auroral activity was strongly enhanced, and to Saturn, where it activated the unusual polar source. This establishes that shocks retain their properties and their ability to trigger planetary auroral activity throughout the Solar System. Our results also reveal differences in the planetary auroral responses on the passing shock, especially in their latitudinal and local time dependences. PMID- 15525982 TI - High-energy particle acceleration in the shell of a supernova remnant. AB - A significant fraction of the energy density of the interstellar medium is in the form of high-energy charged particles (cosmic rays). The origin of these particles remains uncertain. Although it is generally accepted that the only sources capable of supplying the energy required to accelerate the bulk of Galactic cosmic rays are supernova explosions, and even though the mechanism of particle acceleration in expanding supernova remnant (SNR) shocks is thought to be well understood theoretically, unequivocal evidence for the production of high energy particles in supernova shells has proven remarkably hard to find. Here we report on observations of the SNR RX J1713.7 - 3946 (G347.3 - 0.5), which was discovered by ROSAT in the X-ray spectrum and later claimed as a source of high energy gamma-rays of TeV energies (1 TeV = 10(12) eV). We present a TeV gamma-ray image of the SNR: the spatially resolved remnant has a shell morphology similar to that seen in X-rays, which demonstrates that very-high-energy particles are accelerated there. The energy spectrum indicates efficient acceleration of charged particles to energies beyond 100 TeV, consistent with current ideas of particle acceleration in young SNR shocks. PMID- 15525984 TI - Optically programmable electron spin memory using semiconductor quantum dots. AB - The spin of a single electron subject to a static magnetic field provides a natural two-level system that is suitable for use as a quantum bit, the fundamental logical unit in a quantum computer. Semiconductor quantum dots fabricated by strain driven self-assembly are particularly attractive for the realization of spin quantum bits, as they can be controllably positioned, electronically coupled and embedded into active devices. It has been predicted that the atomic-like electronic structure of such quantum dots suppresses coupling of the spin to the solid-state quantum dot environment, thus protecting the 'spin' quantum information against decoherence. Here we demonstrate a single electron spin memory device in which the electron spin can be programmed by frequency selective optical excitation. We use the device to prepare single electron spins in semiconductor quantum dots with a well defined orientation, and directly measure the intrinsic spin flip time and its dependence on magnetic field. A very long spin lifetime is obtained, with a lower limit of about 20 milliseconds at a magnetic field of 4 tesla and at 1 kelvin. PMID- 15525985 TI - Fire-induced erosion and millennial-scale climate change in northern ponderosa pine forests. AB - Western US ponderosa pine forests have recently suffered extensive stand replacing fires followed by hillslope erosion and sedimentation. These fires are usually attributed to increased stand density as a result of fire suppression, grazing and other land use, and are often considered uncharacteristic or unprecedented. Tree-ring records from the past 500 years indicate that before Euro-American settlement, frequent, low-severity fires maintained open stands. However, the pre-settlement period between about ad 1500 and ad 1900 was also generally colder than present, raising the possibility that rapid twentieth century warming promoted recent catastrophic fires. Here we date fire-related sediment deposits in alluvial fans in central Idaho to reconstruct Holocene fire history in xeric ponderosa pine forests and examine links to climate. We find that colder periods experienced frequent low-severity fires, probably fuelled by increased understory growth. Warmer periods experienced severe droughts, stand replacing fires and large debris-flow events that comprise a large component of long-term erosion and coincide with similar events in sub-alpine forests of Yellowstone National Park. Our results suggest that given the powerful influence of climate, restoration of processes typical of pre-settlement times may be difficult in a warmer future that promotes severe fires. PMID- 15525986 TI - Contrasting origins of the upper mantle revealed by hafnium and lead isotopes from the Southeast Indian Ridge. AB - The origin of the isotopic signature of Indian mid-ocean ridge basalts has remained enigmatic, because the geochemical composition of these basalts is consistent either with pollution from recycled, ancient altered oceanic crust and sediments, or with ancient continental crust or lithosphere. The radiogenic isotopic signature may therefore be the result of contamination of the upper mantle by plumes containing recycled altered ancient oceanic crust and sediments, detachment and dispersal of continental material into the shallow mantle during rifting and breakup of Gondwana, or contamination of the upper mantle by ancient subduction processes. The identification of a process operating on a scale large enough to affect major portions of the Indian mid-ocean ridge basalt source region has been a long-standing problem. Here we present hafnium and lead isotope data from across the Indian-Pacific mantle boundary at the Australian-Antarctic discordance region of the Southeast Indian Ridge, which demonstrate that the Pacific and Indian upper mantle basalt source domains were each affected by different mechanisms. We infer that the Indian upper-mantle isotope signature in this region is affected mainly by lower continental crust entrained during Gondwana rifting, whereas the isotope signature of the Pacific upper mantle is influenced predominantly by ocean floor subduction-related processes. PMID- 15525987 TI - The origin of the internal nostril of tetrapods. AB - The choana, a unique 'internal nostril' opening from the nasal sac into the roof of the mouth, is a key part of the tetrapod (land vertebrate) respiratory system. It was the first component of the tetrapod body plan to evolve, well before the origin of limbs, and is therefore crucial to our understanding of the beginning of the fish-tetrapod transition. However, there is no consensus on the origin of the choana despite decades of heated debate; some have claimed that it represents a palatally displaced external nostril, but others have argued that this is implausible because it implies breaking and rejoining the maxillary-premaxillary dental arcade and the maxillary branch of nerve V. The fossil record has not resolved the dispute, because the choana is fully developed in known tetrapod stem-group members. Here we present new material of Kenichthys, a 395-million year-old fossil fish from China, that provides direct evidence for the origin of the choana and establishes its homology: it is indeed a displaced posterior external nostril that, during a brief transitional stage illustrated by Kenichthys, separated the maxilla from the premaxilla. PMID- 15525988 TI - Ecosystem remodelling among vertebrates at the Permian-Triassic boundary in Russia. AB - The mass extinction at the Permian-Triassic boundary, 251 million years (Myr) ago, is accepted as the most profound loss of life on record. Global data compilations indicate a loss of 50% of families or more, both in the sea and on land, and these figures scale to a loss of 80-96% of species, based on rarefaction analyses. This level of loss is confirmed by local and regional-scale studies of marine sections, but the terrestrial record has been harder to analyse in such close detail. Here we document the nature of the event in Russia in a comprehensive survey of 675 specimens of amphibians and reptiles from 289 localities spanning 13 successive geological time zones in the South Urals basin. These changes in diversity and turnover cannot be explained simply by sampling effects. There was a profound loss of genera and families, and simplification of ecosystems, with the loss of small fish-eaters and insect-eaters, medium and large herbivores and large carnivores. Faunal dynamics also changed, from high rates of turnover through the Late Permian period to greater stability at low diversity through the Early Triassic period. Even after 15 Myr of ecosystem rebuilding, some guilds were apparently still absent-small fish-eaters, small insect-eaters, large herbivores and top carnivores. PMID- 15525989 TI - Long-term decline in krill stock and increase in salps within the Southern Ocean. AB - Antarctic krill (Euphausia superba) and salps (mainly Salpa thompsoni) are major grazers in the Southern Ocean, and krill support commercial fisheries. Their density distributions have been described in the period 1926-51, while recent localized studies suggest short-term changes. To examine spatial and temporal changes over larger scales, we have combined all available scientific net sampling data from 1926 to 2003. This database shows that the productive southwest Atlantic sector contains >50% of Southern Ocean krill stocks, but here their density has declined since the 1970s. Spatially, within their habitat, summer krill density correlates positively with chlorophyll concentrations. Temporally, within the southwest Atlantic, summer krill densities correlate positively with sea-ice extent the previous winter. Summer food and the extent of winter sea ice are thus key factors in the high krill densities observed in the southwest Atlantic Ocean. Krill need the summer phytoplankton blooms of this sector, where winters of extensive sea ice mean plentiful winter food from ice algae, promoting larval recruitment and replenishing the stock. Salps, by contrast, occupy the extensive lower-productivity regions of the Southern Ocean and tolerate warmer water than krill. As krill densities decreased last century, salps appear to have increased in the southern part of their range. These changes have had profound effects within the Southern Ocean food web. PMID- 15525990 TI - Baf60c is essential for function of BAF chromatin remodelling complexes in heart development. AB - Tissue-specific transcription factors regulate several important aspects of embryonic development. They must function in the context of DNA assembled into the higher-order structure of chromatin. Enzymatic complexes such as the Swi/Snf like BAF complexes remodel chromatin to allow the transcriptional machinery access to gene regulatory elements. Here we show that Smarcd3, encoding Baf60c, a subunit of the BAF complexes, is expressed specifically in the heart and somites in the early mouse embryo. Smarcd3 silencing by RNA interference in mouse embryos derived from embryonic stem cells causes defects in heart morphogenesis that reflect impaired expansion of the anterior/secondary heart field, and also results in abnormal cardiac and skeletal muscle differentiation. An intermediate reduction in Smarcd3 expression leads to defects in outflow tract remodelling reminiscent of human congenital heart defects. Baf60c overexpressed in cell culture can mediate interactions between cardiac transcription factors and the BAF complex ATPase Brg1, thereby potentiating the activation of target genes. These results reveal tissue-specific and dose-dependent roles for Baf60c in recruiting BAF chromatin remodelling complexes to heart-specific enhancers, providing a novel mechanism to ensure transcriptional regulation during organogenesis. PMID- 15525991 TI - A faux 3'-UTR promotes aberrant termination and triggers nonsense-mediated mRNA decay. AB - Nonsense-mediated messenger RNA decay (NMD) is triggered by premature translation termination, but the features distinguishing premature from normal termination are unknown. One model for NMD suggests that decay-inducing factors bound to mRNAs during early processing events are routinely removed by elongating ribosomes but remain associated with mRNAs when termination is premature, triggering rapid turnover. Recent experiments challenge this notion and suggest a model that posits that mRNA decay is activated by the intrinsically aberrant nature of premature termination. Here we use a primer extension inhibition (toeprinting) assay to delineate ribosome positioning and find that premature translation termination in yeast extracts is indeed aberrant. Ribosomes encountering premature UAA or UGA codons in the CAN1 mRNA fail to release and, instead, migrate to upstream AUGs. This anomaly depends on prior nonsense codon recognition and is eliminated in extracts derived from cells lacking the principal NMD factor, Upf1p, or by flanking the nonsense codon with a normal 3' untranslated region (UTR). Tethered poly(A)-binding protein (Pab1p), used as a mimic of a normal 3'-UTR, recruits the termination factor Sup35p (eRF3) and stabilizes nonsense-containing mRNAs. These findings indicate that efficient termination and mRNA stability are dependent on a properly configured 3'-UTR. PMID- 15525992 TI - Insight into steroid scaffold formation from the structure of human oxidosqualene cyclase. AB - In higher organisms the formation of the steroid scaffold is catalysed exclusively by the membrane-bound oxidosqualene cyclase (OSC; lanosterol synthase). In a highly selective cyclization reaction OSC forms lanosterol with seven chiral centres starting from the linear substrate 2,3-oxidosqualene. Valuable data on the mechanism of the complex cyclization cascade have been collected during the past 50 years using suicide inhibitors, mutagenesis studies and homology modelling. Nevertheless it is still not fully understood how the enzyme catalyses the reaction. Because of the decisive role of OSC in cholesterol biosynthesis it represents a target for the discovery of novel anticholesteraemic drugs that could complement the widely used statins. Here we present two crystal structures of the human membrane protein OSC: the target protein with an inhibitor that showed cholesterol lowering in vivo opens the way for the structure-based design of new OSC inhibitors. The complex with the reaction product lanosterol gives a clear picture of the way in which the enzyme achieves product specificity in this highly exothermic cyclization reaction. PMID- 15525993 TI - Membrane structure and interactions with protein and DNA in bacteriophage PRD1. AB - Membranes are essential for selectively controlling the passage of molecules in and out of cells and mediating the response of cells to their environment. Biological membranes and their associated proteins present considerable difficulties for structural analysis. Although enveloped viruses have been imaged at about 9 A resolution by cryo-electron microscopy and image reconstruction, no detailed crystallographic structure of a membrane system has been described. The structure of the bacteriophage PRD1 particle, determined by X-ray crystallography at about 4 A resolution, allows the first detailed analysis of a membrane containing virus. The architecture of the viral capsid and its implications for virus assembly are presented in the accompanying paper. Here we show that the electron density also reveals the icosahedral lipid bilayer, beneath the protein capsid, enveloping the viral DNA. The viral membrane contains about 26,000 lipid molecules asymmetrically distributed between the membrane leaflets. The inner leaflet is composed predominantly of zwitterionic phosphatidylethanolamine molecules, facilitating a very close interaction with the viral DNA, which we estimate to be packaged to a pressure of about 45 atm, factors that are likely to be important during membrane-mediated DNA translocation into the host cell. In contrast, the outer leaflet is enriched in phosphatidylglycerol and cardiolipin, which show a marked lateral segregation within the icosahedral asymmetric unit. In addition, the lipid headgroups show a surprising degree of order. PMID- 15525994 TI - Home and away. PMID- 15525995 TI - High road to Scotland. PMID- 15525996 TI - Pharmacological and behavioral characteristics of interactions between vigabatrin and conventional antiepileptic drugs in pentylenetetrazole-induced seizures in mice: an isobolographic analysis. AB - To characterize the anticonvulsant effects and types of interactions exerted by mixtures of vigabatrin (VGB) and conventional antiepileptic drugs (valproate (VPA), ethosuximide (ESM), phenobarbital (PB), and clonazepam (CZP)) in pentylenetetrazole (PTZ)-induced seizures in mice, the isobolographic analysis for three fixed-ratio combinations of 1 : 3, 1 : 1, and 3 : 1 was used. The adverse-effect profile of the combinations tested, at the doses corresponding to their median effective doses (ED(50)) at the fixed-ratio of 1 : 1 against PTZ induced seizures, was determined by the chimney (motor performance), step-through passive avoidance (long-term memory), pain threshold (pain sensitivity), and Y maze (general explorative locomotor activity) tests in mice. Additionally, the observed isobolographic interactions were verified in terms of a pharmacokinetic interaction existence. VGB combined with PB or ESM exerted supra-additive (synergistic) interactions against the clonic phase of PTZ-induced seizures, which was associated with the increment of PB or ESM concentrations in the brains of examined animals. The remaining combinations tested (ie VGB+VPA and VGB+CZP) occurred additive in the PTZ test, which was associated with no significant changes in the brain concentrations of VPA and CZP. None of the examined combinations exerted motor impairment in the chimney test in mice. In the standard variant of passive avoidance task (current of 0.6 mA; 2 s of stimulus duration), the combinations of VGB+CZP and VGB+VPA significantly affected long term memory in mice. Moreover, VGB in a dose-dependent manner lengthened the latency to the first pain reaction in the pain threshold test in mice. The modified variant of step-through passive avoidance task (current of 0.6 mA; stimulus duration based on the latency from the pain threshold test) revealed no significant changes in the long-term memory of animals for the combinations of VGB+VPA and VGB+CZP; so the observed effects in the standard variant of passive avoidance task were a result of the antinociceptive effects produced by VGB. In the Y-maze test, VGB also, in a dose-dependent manner, increased the general explorative locomotor activity of the animals tested. Similarly, the total number of arm entries in the Y-maze was significantly increased for the combinations of VGB+CZP and VGB+ESM, but not for VGB+PB and VGB+VPA. The application of VGB in combination with PB, ESM, CZP, and VPA suppressed the clonic phase of PTZ-induced seizures, having no harmful or deleterious effects on behavioral functioning of the animals tested, which might be advantageous in further clinical practice. PMID- 15525997 TI - Downregulation of endocannabinoid signaling in the hippocampus following chronic unpredictable stress. AB - Deficits in cognitive functioning and flexibility are seen following both chronic stress and modulation of endogenous cannabinoid (eCB) signaling. Here, we investigated whether alterations in eCB signaling might contribute to the cognitive impairments induced by chronic stress. Chronic stress impaired reversal learning and induced perseveratory behavior in the Morris water maze without significant effect on task acquisition. These cognitive impairments were reversed by exogenous cannabinoid administration, suggesting deficient eCB signaling underlies these phenomena. In line with this hypothesis, chronic stress downregulated CB1 receptor expression and significantly reduced the content of the endocannabinoid 2-arachidonylglycerol within the hippocampus. CB1 receptor density and 2-arachidonylglycerol content were unaffected in the limbic forebrain. These data suggest that stress-induced downregulation of hippocampal eCB signaling contributes to problems in behavioral flexibility and could play a role in the development of perseveratory and ruminatory behaviors in stress related neuropsychiatric disorders. PMID- 15525998 TI - A double-blind, placebo-controlled trial of modafinil for cocaine dependence. AB - Despite years of active research, there are still no approved medications for the treatment of cocaine dependence. Modafinil is a glutamate-enhancing agent that blunts cocaine euphoria under controlled conditions, and the current study assessed whether modafinil would improve clinical outcome in cocaine-dependent patients receiving standardized psychosocial treatment. This was a randomized, double-blind, placebo-controlled trial conducted at a university outpatient center (from 2002 to 2003) on a consecutive sample of 62 (predominantly African American) cocaine-dependent patients (aged 25-63) free of significant medical and psychiatric conditions. After screening, eligible patients were randomized to a single morning dose of modafinil (400 mg), or matching placebo tablets, for 8 weeks while receiving manual-guided, twice-weekly cognitive behavioral therapy. The primary efficacy measure was cocaine abstinence based on urine benzoylecgonine levels. Secondary measures were craving, cocaine withdrawal, retention, and adverse events. Modafinil-treated patients provided significantly more BE-negative urine samples (p=0.03) over the 8-week trial when compared to placebos, and were more likely to achieve a protracted period (> or =3 weeks) of cocaine abstinence (p=0.05). There were no serious adverse events, and none of the patients failed to complete the study as a result of adverse events. This study provides preliminary evidence, which should be confirmed by a larger study, that modafinil improves clinical outcome when combined with psychosocial treatment for cocaine dependence. PMID- 15525999 TI - Increased attributable risk related to a functional mu-opioid receptor gene polymorphism in association with alcohol dependence in central Sweden. AB - The mu-opioid receptor (MOR), through its effects on reward and stress responsivity, modulates alcohol intake in both animal and human laboratory studies. We have previously demonstrated that the frequently occurring A118G single-nucleotide polymorphism (SNP) in exon 1 of the MORgene (OPRM1), which encodes an amino-acid substitution, is functional and receptors encoded by the variant 118G allele bind the endogenous opioid peptide beta-endorphin with three fold greater affinity than prototype receptors. Other groups subsequently reported that this variant alters stress-responsivity in normal volunteers and also increases the therapeutic response to naltrexone (a mu-preferring opioid antagonist) in the treatment of alcohol dependence. We compared frequencies of genotypes containing an 118G allele in 389 alcohol-dependent individuals and 170 population-based controls without drug or alcohol abuse or dependence. The A118G SNP was present in the Hardy-Weinberg equilibrium with an overall frequency of the 118G allele of 10.9%. There was a significant overall association between genotypes with an 118G allele and alcohol dependence (p=0.0074). The attributable risk for alcohol dependence in subjects with an 118G allele was 11.1%. There was no difference in A118G genotype between type 1 and type 2 alcoholics. In central Sweden, the functional variant 118G allele in exon 1 of OPRM1 is associated with an increased attributable risk for alcohol dependence. PMID- 15526000 TI - Evaluation of the reinforcing effects of monoamine reuptake inhibitors under a concurrent schedule of food and i.v. drug delivery in rhesus monkeys. AB - Most medications prescribed for attention-deficit-hyperactivity disorder are psychomotor stimulants with reinforcing effects in laboratory animals (eg methylphenidate). The present studies were conducted to evaluate the reinforcing effects of the recently approved medication atomoxetine in monkeys trained to 'choose' between automated deliveries of either an i.v. injection or food. Rhesus monkeys were trained to lever-press under concurrent schedules of reinforcement; responses on one lever resulted in an injection of either saline or drug, and responses on the alternative lever resulted in food delivery. Data were collected on four measures: response rates, percentage of total responses occurring on the injection-lever (% ILR), number of injections earned, and number of food pellets earned. Dose-effect functions were determined for cocaine (0.003-0.3 mg/kg/inj), methylphenidate (0.003-0.1 mg/kg/inj), amphetamine (0.003-0.1 mg/kg/inj), atomoxetine (0.01-0.3 mg/kg/inj), and desipramine (0.03-1.0 mg/kg/inj) using a double alternation schedule of saline and drug availability. Results indicate that the distribution of behavior changed according to the drug and dose available for self-injection. Saline availability was typically associated with high rates of food-maintained responding. The % ILR increased from 3+/-1% when saline was available to >90% when >0.03 mg/kg/inj of cocaine, methylphenidate or d-amphetamine was available. However, no dose of atomoxetine or desipramine maintained self-administration behavior on the injection-lever. The number of food pellets earned per session decreased as the dose of each drug increased, indicative of behavioral activity with all five drugs. The reinforcing effects of cocaine, methylphenidate, and d-amphetamine in these studies are consistent with previous findings in nonhuman primates and with their documented abuse liability. The absence of reinforcing effects of atomoxetine support the view that, like desipramine, it has no evident abuse potential. PMID- 15526001 TI - Is there an interaction between interleukin-10 and interleukin-22? AB - Interleukin(IL)-10 and IL-22 are structurally related cytokines. Their heterodimeric receptors consist of the cytokine-specific chains IL-10R1 and IL 22R1, respectively, and the common chain IL-10R2. This study focused on the question of whether IL-10 modulates IL-22 effects and vice versa. This question is important because IL-10 and IL-22 exert anti- and proinflammatory effects, respectively, and, as we show here, are simultaneously present in both systemic and local inflammation. The revealed lacking concomitance of IL-10R1 and IL-22R1 on identical cells excluded any possible interaction between IL-10 and IL-22 apart from the competition for IL-10R2. To study this competition, monocytes and hepatocytes were chosen. The dependence of the cytokine action on IL-10R2 was verified. Interestingly, no influence of IL-22 on IL-10 effects was observed. The same was true when IL-22 was used in complex with IL-22-binding protein. Similarly, no influence of IL-10 was found on IL-22 action. This missing competition seemed to be due to a lack of binding between IL-10R2 and the native cytokines in the absence of their corresponding R1 chain. However, IL-10R2 interacted with defined IL-10- and IL-22-derived peptides supporting the hypothesis that cytokine binding to its corresponding R1 chain creates a binding site on this cytokine for IL-10R2. PMID- 15526002 TI - Characterization of the divergent eosinophil ribonuclease, mEar 6, and its expression in response to Schistosoma mansoni infection in vivo. AB - The eosinophil-associated ribonucleases (Ears) are rapidly evolving proteins found in multigene clusters that are unique to each rodent species. Of the 15 independent genes in the Mus musculus cluster, only mEars 1 and 2 are expressed at significant levels at homeostasis. Here we characterize the expression of mEar 6 in the liver and spleen in mice in response to infection with the helminthic parasite, Schistosoma mansoni. Interestingly, expression of mEar 6 is not directly related to the elevated levels of serum IL-5 or tissue eosinophilia characteristic of this disease, as no mEar 6 transcripts were detected in the liver or the spleen from uninfected IL-5-transgenic mice. The coding sequence of mEar 6 has diverged under positive selection pressure (K(a)/K(s) > 1.0) and has a unique unpaired cysteine near the carboxy-terminus of the protein. The high catalytic efficiency of recombinant mEar 6 (k(cat)/K(m) = 0.9 x 10(6)/M/s) is similar to that of the cluster's closest human ortholog, eosinophil-derived neurotoxin (EDN/RNase 2). In summary, we have identified mEar 6 as one of only two RNase A superfamily ribonucleases known to be expressed specifically in response to pathophysiologic stress in vivo. PMID- 15526003 TI - A functional polymorphism (1858C/T) in the PTPN22 gene is linked and associated with type I diabetes in multiplex families. AB - Type I diabetes (T1D) is a complex disorder, which arises from the autoimmune destruction of the insulin-secreting beta cells of the pancreas leading to a life long dependence on exogenous insulin. A recent study of T1D cases and controls provided evidence for association between an allele of a functional single nucleotide polymorphism (SNP) in the PTPN22 gene and T1D. In the current study, this SNP was genotyped in a collection of 406 multiplex T1D families. Significant evidence of the combined presence of association and linkage to T1D was obtained (P = 2.5 x 10(-5)). Linkage studies in subsets of families defined by PTPN22 SNP genotypes suggest possible interaction with loci on chromosomes 3 and 21. Previous genome scans in this collection of T1D families, and others, have not yielded significant evidence of linkage in the region of the PTPN22 locus. However, the highly significant evidence of allelic association suggests that variation at, or near, this functional SNP contributes to the risk of T1D. PMID- 15526004 TI - Genetic risk factors for infection in patients with early rheumatoid arthritis. AB - We analyzed clinical and genetic factors contributing to infections in 457 subjects with early rheumatoid arthritis (RA) enrolled in a prospective, 1-year clinical trial of methotrexate and the TNF inhibitor etanercept. Subjects were genotyped for the following single nucleotide polymorphisms (SNPs): (TNF -308, 238, and + 488); lymphotoxin-alpha (LTA) (LTA + 249, + 365, and + 720); and Fc gamma receptors FCGR2A 131 H/R; FCGR3A 176 F/V; and FCGR3B NA 1/2 and genotypes were correlated with infections. At least one URI was noted in 52% of subjects (99/191) with the NA2/NA2 genotype of the neutrophil-specific FCGR3B gene, compared to 42% (77/181) of those with the NA1/NA2 genotype and 39% (23/59) of those with the NA1/NA1 genotype (P = 0.038). Urinary tract infection (UTI) was associated with the TNF -238 A (odds ratio(OR) 2.56, 95% confidence interval (CI) 1.05-6.25) and LTA +365 C (OR 1.73, 95% CI 1.07-2.79) alleles, and marginally with the FCGR3A F allele (OR 1.72, 95% CI 0.99-3.00). There was a striking linear correlation between UTI and the number of risk alleles defined by these three SNPs (P < 0.001), suggesting an additive effect on susceptibility. These findings have important implications for the role of genetics in susceptibility to bacterial and viral infections. PMID- 15526005 TI - TNF and TNFR polymorphisms in severe sepsis and septic shock: a prospective multicentre study. AB - Tumour necrosis factor (TNF) is an important pro-inflammatory cytokine produced in sepsis. Studies examining the association of individual TNF single nucleotide polymorphisms with sepsis have produced conflicting results. This study investigated whether common polymorphisms of the TNF locus and the two receptor genes, TNFRSF1A and TNFRSF1B, influence circulating levels of encoded proteins, and whether individual polymorphisms or extended haplotypes of these genes are associated with susceptibility, severity of illness or outcome in adult patients with severe sepsis or septic shock. A total of 213 Caucasian patients were recruited from eight intensive care units (ICU) in the UK and Australia. Plasma levels of TNF (P = 0.02), sTNFRSF1A (P = 0.005) and sTNFRSF1B (P = 0.01) were significantly higher in those who died on ICU compared to those who survived. There was a positive correlation between increasing soluble receptor levels and organ dysfunction (increasing SOFA score) (sTNFRSF1A R = 0.51, P < 0.001; sTNFRSF1B R = 0.53, P < 0.001), and in particular with the degree of renal dysfunction. In this study, there were no significant associations between the selected candidate TNF or TNF receptor polymorphisms, or their haplotypes, and susceptibility to sepsis, illness severity or outcome. The influence of polymorphisms of the TNF locus on susceptibility to, and outcome from sepsis remains uncertain. PMID- 15526006 TI - Immunotherapy with plasmid DNA encoding mycobacterial hsp65 in association with chemotherapy is a more rapid and efficient form of treatment for tuberculosis in mice. AB - Tuberculosis (TB) remains a threat for public health, killing around 3 million people a year. Despite the fact that most cases can be cured with antibiotics, the treatment is long and patients relapse if chemotherapy is not continued for at least 6 months. Thus, a better characterization of the working principles of the immune system in TB and identification of new immunotherapeutic products for the development of shorter regimens of treatment are essential to achieve an effective management of this disease. In the present work, we demonstrate that immunotherapy with a plasmid DNA encoding the Mycobacterium leprae 65 kDa heat shock protein (hsp65) in order to boost the efficiency of the immune system, is a valuable adjunct to antibacterial chemotherapy to shorten the duration of treatment, improve the treatment of latent TB infection and be effective against multidrug-resistant bacilli (MDR-TB). We also showed that the use of DNA-hsp65 alone or in combination with other drugs influence the pathway of the immune response or other types of inflammatory responses and should augment our ability to alter the course of immune response/inflammation as needed, evidencing an important target for immunization or drug intervention. PMID- 15526007 TI - Mesothelin-mediated targeting of adenoviral vectors for ovarian cancer gene therapy. AB - Adenoviruses (Ads) are efficient gene transfer vehicles, but Ad-mediated gene therapy for ovarian cancer remains limited in vivo by inefficient and nonspecific gene transfer. Mesothelin (MSLN), a cell surface glycoprotein, is overexpressed in ovarian cancer but not in normal tissues except mesothelial cells. Therefore, MSLN is an attractive candidate for transcriptional and transductional targeting in the context of ovarian cancer gene therapy. We evaluated the expression of MSLN mRNA and MSLN surface protein in ovarian cancer cells. Ads containing the MSLN promoter driving reporter gene expression were created and tested in ovarian cancer cell lines and purified ovarian cancer cells isolated from patients. To evaluate transductional targeting, we used an Ad vector containing an Fc-binding domain within the fiber protein, which served as a docking domain for binding with anti-MSLN immunoglobulins. Both RT-PCR and flow cytometry revealed high MSLN gene and protein expression in ovarian cancer cells. The MSLN promoter was activated in ovarian cancer cells, but showed significantly reduced activity in normal control cells. Transductional targeting of Ads via anti-MSLN antibody increased transgene expression in ovarian cancer cells. This report describes the use of MSLN for transcriptional as well as transductional targeting strategies for ovarian cancer gene therapy. PMID- 15526008 TI - Long-term efficacy and compliance of MUSE for erectile dysfunction following radical prostatectomy: SHIM (IIEF-5) analysis. AB - Baseline and follow-up data of 54 patients from a single surgical series (1998 2001), who used medicated urethral system for erection (MUSE) for the erectile dysfunction (ED) associated with radical prostatectomy (RP), were obtained. Patients were surveyed using the abridged five-item version of the International Index of Erectile Function (IIEF) questionnaire, commonly referred to as the Sexual Health Inventory of Men (SHIM), to determine presence and severity of ED and efficacy of ED treatment modalities. The mean patient age was 63.7+/-5.6 y and the mean follow-up period was 2.3+/-1.2 y. All patients experienced ED for at least 6 months after their surgery before starting MUSE therapy. Overall, 55% of the patients achieved and maintained erections sufficient for sexual intercourse while on MUSE and 48% continued long-term therapy with a mean use of 2.32+/-1.2 y. The mean presurgery SHIM score in these patients was 19.2+/-1.3, which decreased to 5.2+/-0.5 after surgery and increased to 16.3+/-1.3 after MUSE treatment. A total of 28 patients (52%) discontinued treatment after a mean use of 8+/-1.4 months. The reasons for discontinuation were insufficient erections (n = 16, mean SHIM score of 10.5+/-4.4), switch to other ED therapies (n = 4), natural return of erections (n = 4) and urethral pain and burning (n = 4). Excluding the patients (n = 8) who preferred other therapies and return of natural erections, the compliance to MUSE was 63%. There were no significant differences in the IIEF-5 responses between the patients who had a nerve-sparing technique (n=34) and those who did not (n = 20) or among patients who used different doses (250, 500 or 1000 microg) of MUSE. The results of the current trial indicate that MUSE is a successful treatment option in RP patients with established ED. It appears that a post-treatment SHIM score of > or = 16 defines a successful outcome with MUSE therapy. PMID- 15526009 TI - Vasoactive intestinal polypeptide (VIP) is not an androgen-dependent neuromediator of penile erection. AB - The effects of castration on vasoactive intestinal polypeptide (VIP) immunostaining in human corpus cavernosum (CC) and the relationship between VIP immunostaining and erectile function were studied in patients with localised prostate cancer who had (Group 1 = castrated) or had not (Group 2 = control) undergone 3-month neoadjuvant chemical castration before radical prostatectomy. Evaluation of erectile function included medical and sexual history, physical examination, and measurement of total serum testosterone. CC biopsies were taken at the end of radical prostatectomy and samples immunostained with anti-human VIP antibody. Specific staining was quantified by image analysis and expressed in arbitrary units (AU). Chemical castration induced erectile function deterioration in 70% of patients due to loss of sexual interest and confidence in the ability of having an erection rather than reduced ability of obtaining sexually induced erections. Average VIP content was 34.5 AU in Group 1 and 39 AU in Group 2 and this difference was not statistically significant. Chemical castration does not influence VIP immunostaining of human CC, suggesting that VIP is not an androgen dependent neuromediator of penile erection and that it can be responsible for sexually induced erections in castrated patients. PMID- 15526010 TI - Prothrombotic risk factors in infants of diabetic mothers. AB - BACKGROUND: Infants of diabetic mothers (IDMs) are at an increased risk for thromboembolic disease. The mechanism(s) to explain this association is unclear. We hypothesized that the pathophysiology of thrombosis in IDMs is multifactorial and likely involves interactions among genetic and acquired factors affecting the procoagulant, anticoagulant and fibrinolytic pathways. OBJECTIVE: To compare the prevalence of common prothrombotic risk factors in a cohort of IDMs to a matched control group. PATIENTS/METHODS: Full-term infants born to mothers with diet controlled (A1-IDM) (N=17), insulin requiring diabetes (ID-IDM) (N=20) and healthy term infants (controls) (N=20) matched for mode of delivery had cord blood collected at delivery. Samples were analyzed for the following: factor V Leiden (FVL), prothrombin 20210A (P20210A), methylenetetrahydrofolate reductase C677 T (MTHFR), Factor VIII (FVIII), Protein C (PC), Lipoprotein(a) (Lp(a)) and plasminogen activator inhibitor-1 (PAI-1). RESULTS: None of the infants had a clinically apparent thrombotic event. IDM mothers and their infants were clinically similar to controls except for a higher prevalence of hypoglycemia (30 vs 0%; p=0.005). There was no significant difference in the prevalence of the common genetic risk factors (FVL, P20210A, MTHFR) FVIII, or PAI-1 levels. Elevated Lp(a) levels were seen more frequently in IDMs than Controls (40 vs 20%) but this difference was not statistically significant. The PC activity (%) was significantly decreased in the IDM group compared to controls, 35+/-12 vs 44+/-9 (p<0.005). A1-IDM had lower PC activity compared to ID-IDM (p=0.05) and controls (p=0.001). CONCLUSIONS: PC deficiency is likely one mechanism to explain thrombosis in IDMs. PMID- 15526011 TI - The effect of intermittent maternal fasting on computerized fetal heart tracing. AB - OBJECTIVE: To determine effects of maternal fasting on antepartum computerized fetal heart tracing analysis. STUDY DESIGN: This was a cross-sectional study of two groups of healthy pregnant women who were recruited, a fasting and a nonfasting control group. Each pregnant woman gave a blood sample, and had a computerized fetal heart tracing. RESULTS: A total of 124 pregnant women were studied. The mean hours since the last oral intake were significantly different between the two groups (p=0.003). Fetuses of fasted mothers had fewer episodes of large acceleration compared to the controls (p=0.001). This difference was significantly associated with maternal appreciation of fetal movement (p=0.003). CONCLUSION: The number of large accelerations in computerized fetal heart tracing is decreased in pregnant women abstaining from food and water. PMID- 15526012 TI - Effect of short-term erythropoietin therapy in anemic premature infants. AB - OBJECTIVE: To determine the effectiveness of a 10-day subcutaneous erythropoietin (rHuEpo) course of 300 units per kg per dose plus oral iron compared to oral iron alone in anemic infants during their convalescent phase of illness. STUDY DESIGN: Prospective, randomized trial performed at a 40-bed, teaching, referral, level III, neonatal intensive care unit. Infants with a gestational age at birth of less than 32 weeks, hematocrit of less than or equal to 28% with a corrected reticulocyte count of less than or equal to 5%, postconceptual age of less than 48 weeks or 5 months chronological age, and a diagnosis of anemia of prematurity were considered for inclusion. Major outcome parameters included hematocrit, corrected reticulocyte count and red cell transfusion requirements. RESULTS: A total of 60 infants were enrolled (n=30 per group). Infants randomized to rHuEpo had a significantly higher post-treatment hematocrit and corrected reticulocyte count than infants in the iron only group (p<0.001). There was a trend towards fewer red cell requirements in the rHuEpo group. CONCLUSIONS: The rHuEpo regimen studied here was associated with an acute improvement in hematocrit and corrected reticulocyte counts. This study did not demonstrate a statistically significant decrease in transfusion therapy, in part related to increased subsequent use of rHuEpo in the control group. Taken together, these data demonstrate that this regimen can effectively treat anemia in convalescent premature infants. PMID- 15526013 TI - Prevalence of low cortisol values in term and near-term infants with vasopressor resistant hypotension. AB - OBJECTIVE: To determine the prevalence of low cortisol values and to evaluate clinical responses to hydrocortisone in ill term and near-term newborns. STUDY DESIGN: Retrospective cohort study including infants >or=35 weeks gestational age who were mechanically ventilated, received vasopressor therapy, and had a cortisol concentration obtained for evaluation of vasopressor-resistant hypotension. In those infants treated with hydrocortisone, physiologic responses were evaluated and correlated with cortisol values (<15 vs >or=15 microg/dl). RESULTS: A total of 32 infants had cortisol values obtained; 18 (56%) were <15 microg/dl. In all, 21 infants were treated with hydrocortisone of whom 13 had cortisol values <15 microg/dl. These 13 infants showed decreased heart rate, dopamine support and fluid bolus requirements after treatment, compared to infants with values >or=15 microg/dl (n=8). CONCLUSIONS: A significant number of hypotensive, mechanically ventilated infants have evidence of inadequate adrenal function. Hydrocortisone therapy resulted in hemodynamic stabilization in this population. PMID- 15526014 TI - Severe thrombocytopenia predicts outcome in neonates with necrotizing enterocolitis. AB - OBJECTIVE: Necrotizing enterocolitis (NEC) is a common and serious gastrointestinal disorder that predominately affects premature infants. Few prognostic indices are available to guide physicians through the expected course of the disease. We hypothesized that the degree and timing of onset of severe thrombocytopenia (platelet count <100,000/mm(3)) would be a predictor of adverse outcome and an indication for surgical intervention in infants with NEC. STUDY DESIGN: The clinical presentation and outcome of all infants with Bell stage II or III NEC treated at Texas Children's Hospital between 1997 and 2001 were retrospectively reviewed. Patients were stratified into two groups based on the presence (Group1) or absence (Group 2) of severe thrombocytopenia (platelet count <100,000/mm(3)) within 3 days of a diagnosis of NEC. Differences between groups were compared using logistic regression to estimate adjusted odds ratios. RESULTS: A total of 91 infants met inclusion criteria (average birth weight 1288+/-135 g; average gestational age 29.0+/-3.0 weeks). Compared to infants in Group 2, infants in Group 1 were more premature (28.0+/-4.1 vs 30.0+/-4.2 weeks; p=0.02), more likely to have received postnatal steroids (42.5% vs 20.4%; p=0.02), and more likely to require laparotomy for gangrenous bowel (adjusted OR 16.33; p<0. 001). The presence of severe thrombocytopenia was also a predictor of mortality (adjusted OR 6.39; p=0.002) and NEC-related gastrointestinal complications including cholestatic liver disease and short bowel syndrome (adjusted OR 5.47; p=0.006). CONCLUSION: Severe thrombocytopenia within the first 3 days after a diagnosis of NEC suggests a higher likelihood of bowel gangrene, morbidity, and mortality. Prospective studies of infants with early and severe thrombocytopenia may help determine the optimal timing of laparotomy in infants with NEC. PMID- 15526015 TI - Unrelated donor transplantation over the age of 55. Are we merely getting (b)older? PMID- 15526016 TI - Hematopoietic stem-cell transplantation from unrelated donors in elderly patients (age >55 years) with hematologic malignancies: older age is no longer a contraindication when using reduced intensity conditioning. AB - Allogeneic stem cell transplantation (SCT) is a potentially curative approach for patients with hematological malignancies. Reduced-intensity conditioning regimens allow SCT in elderly patients; however, there are only limited data on the feasibility and outcomes of unrelated donor SCT in these patients. In this study, we analyzed, retrospectively, data of 36 patients with various hematological malignancies and median age 58 years (range, 55-66), who were given unrelated donor SCT after reduced-intensity conditioning. The preparative regimen consisted of fludarabine combined with oral busulfan (8 mg/kg, n=8), intravenous busulfan (6.4 mg/kg, n=11), treosulfan (30 g/m(2), n=5) or melphalan (100-150 mg/m(2), n=12). Patients were also given serotherapy, ATG (n=32), or alemtuzumab (n=4). The probabilities of overall survival, disease-free survival, and nonrelapse mortality at 1 year after SCT were 52, 43, and 39%, respectively. Acute graft versus-host disease (GVHD) grade II-IV and chronic GVHD occurred in 31 and 45%, respectively. Multivariable analysis determined that survival rates were higher in patients with chemosensitive disease (HR 4.5), and patients conditioned with intravenous busulfan or treosulfan (HR 3.9). Unrelated donor SCT is feasible in elderly patients, with outcomes that are similar to younger patients. Favorable outcome was observed in patients with myeloid malignancies, and those transplanted in remission and early in the course of disease. Age alone should not be considered a contraindication to unrelated donor SCT. PMID- 15526017 TI - Acute lymphoblastic leukemia after tandem autologous stem cell transplantations for multiple myeloma. PMID- 15526018 TI - A novel approach to identify antigens recognized by CD4 T cells using complement opsonized bacteria expressing a cDNA library. AB - In patients with hematological malignancies receiving HLA-matched stem cell transplantation, T cells specific for minor histocompatibility antigens play a major role in graft rejection, induction of graft-versus-host disease and beneficial graft-versus-leukemia reactivity. Several human minor histocompatibility antigens recognized by T cells have been identified, but only two are presented by HLA class II molecules. In search of an efficient approach to identify antigenic peptides processed through the HLA class II pathway, we constructed a cDNA library in bacteria that were induced to express proteins. Bacteria were opsonized with complement to enforce receptor-mediated uptake by Epstein-Barr virus immortalized B cells that were subsequently used as antigen presenting cells. This approach was validated with an HLA class II-restricted antigen encoded by gene DBY. We were able to identify bacteria expressing DBY diluted into a 300-fold excess of bacteria expressing a nonrelevant gene. Screening of a bacterial library using a DBY-specific CD4 T cell clone resulted in the isolation of several DBY cDNAs. We propose this strategy for a rapid identification of HLA class II-restricted antigenic peptides recognized by CD4 T cells. PMID- 15526023 TI - Upregulation of asparagine synthetase and cell cycle arrest in t(12;21)-positive ALL. PMID- 15526021 TI - A phase I study of 153Sm-EDTMP with fixed high-dose melphalan as a peripheral blood stem cell conditioning regimen in patients with multiple myeloma. AB - Despite response rates of 30% after high-dose chemotherapy with autologous hematopoietic stem cell transplant, patients with multiple myeloma are not cured. 153Samarium ethylenediaminetetramethylenephosphonate (153Sm-EDTMP; Quadramet) is a short-range, beta-emitting therapeutic radiopharmaceutical with avid skeletal uptake. In total, 12 patients were treated with escalating doses of 153Sm-EDTMP (N=3/group; 6, 12, 19.8, and 30 mCi/kg) and a fixed dose of melphalan (200 mg/m(2)). No dose limiting toxicity was seen. To better standardize the marrow compartment radiation dose, the study was modified such that an additional six patients were treated at a targeted absorbed radiation dose to the red marrow of 40 Gy based on a trace labeled infusion 1 week prior to the therapy. Despite rapid elimination of unbound radiopharmaceutical via kidneys and bladder, no episodes of nephrotoxicity, hemorrhagic cystitis, or delayed radiation nephritis were observed with a median follow-up of 31 months (range 8.5-44). Median times to ANC>0.5 and platelet >20 x 10(6)/l were 12 and 11 days, respectively, with no graft failures. Overall response rate was 94% including seven very good partial responses and five complete responses. Addition of 153Sm EDTMP to melphalan conditioning appears to be safe, well-tolerated and worthy of further study. PMID- 15526024 TI - Minor histocompatibility antigen-specific T cells with multiple distinct specificities can be isolated by direct cloning of IFNgamma-secreting T cells from patients with relapsed leukemia responding to donor lymphocyte infusion. AB - Graft-vs-leukemia reactivity after donor lymphocyte infusion (DLI) can be mediated by donor T cells recognizing minor histocompatibility antigens (mHags) on recipient hematopoietic cells. To study the diversity of cells involved in this immune response, hematopoietic cell reactive T cells were directly clonally isolated from peripheral blood of patients entering complete remission after DLI. T cells were briefly stimulated with bone marrow cells from patients pretransplant, and IFNgamma-secreting T cells were directly clonally isolated, and expanded. Cytotoxic T-lymphocyte (CTL) clones from individual patients used multiple distinct HLA-restricting molecules and varied in reactivity against patient-derived normal and/or malignant hematopoietic cells. For each patient, CTL clones specific for known immunodominant mHags as well as distinct unknown mHags were found. Within individual patients, CTL clones using the same HLA restricting element could show differential recognition patterns, indicating further diversity in mHag reactivity. CTL clones from individual patients exhibiting identical specificities could show oligoclonal origin. In conclusion, the direct cloning technique shows that the response to hematopoietic cells after DLI is directed against multiple distinct mHags, including but not limited to known immunodominant mHags, implying that immunotherapy with T cells against multiple mHag specificities may be more effective in eradicating malignant cells. PMID- 15526025 TI - Checkpoint kinase 1 (CHK1) protein and mRNA expression is downregulated in aggressive variants of human lymphoid neoplasms. AB - CHK1: gene encodes for a serine/threonine kinase involved in the regulation of cell cycle progression and DNA damage checkpoints. To determine the role of CHK1 in the pathogenesis of lymphoid neoplasms and its relationship to other DNA damage response genes, we have analyzed the gene status, protein, and mRNA expression in a series of tumors and nonneoplastic lymphoid tissues. CHK1 protein and mRNA expression levels were very low in both reactive tissues and resting lymphoid cells, whereas tumor samples showed a variable pattern of expression related to their proliferative activity. However, seven aggressive tumors showed a dissociate pattern of extremely low or negative protein expression in spite of a high proliferative activity. Four of these tumors were diffuse large B-cell lymphomas (DLCLs) with concordant reduced levels of mRNA, whereas one blastoid mantle cell lymphoma (B-MCL) and two DLCLs had relatively normal levels of mRNA. No gene mutations, deletions, or hypermethylation of the promoter region were detected in any of these cases. In all these tumors ATM, CHK2, and p53 genes were wild type. These findings suggest that CHK1 inactivation in NHLs occurs by loss of protein expression in a subset of aggressive variants alternatively to ATM, CHK2, and p53 alterations. PMID- 15526026 TI - Impact of plasmacytoid dendritic cells on outcome after reduced-intensity conditioning allogeneic stem cell transplantation. AB - The reconstitution of the plasmacytoid dendritic cells (PDCs) compartment might influence outcome after allogeneic stem cell transplantation (allo-SCT). Thus, we investigated the impact of blood PDCs measured at the third month after reduced intensity conditioning (RIC) in 54 patients who received an HLA-identical sibling allo-SCT. The absence of grade II-IV acute graft-versus-host-disease (GVHD) was associated with an improved PDC count at 3 months after RIC-allo-SCT (P=0.003; OR=6.4; 95% CI, 1.9-22). The CD34+ stem cell dose and other lymphoid subsets infused with the allograft did not affect PDC recovery. Although PDC count could not predict death from progression or relapse, patients with a "high" PDC recovery profile had an improved overall survival (OS; P=0.03), in contrast to patients with a "low" PDC recovery profile who had an increased incidence of nonrelapse mortality (GVHD, infections) (P=0.03). The overall incidence of late infections (viral, fungal and bacterial) was significantly higher in the "low" PDC recovery group as compared to the "high" PDC recovery group (59 vs 19%; P=0.002). In a multivariate analysis, only a "high" PDC count was significantly predictive of a decreased risk of death (P=0.04; RR=0.34; 95% CI, 0.12-0.96). Monitoring of PDCs at 3 months after RIC-allo-SCT may be a useful indicator predictor of long-term outcome. PMID- 15526027 TI - Phenotype and function of a CD56+ peripheral blood monocyte. AB - G-CSF primed CD34 cells cultured for 2-3 weeks in IL-2 and stem cell factor generate CD56(high) cells with phenotypic and morphologic features of NK cells, and a novel adherent CD56(low) CD16- population expressing myeloid markers (CD33 and HLA-DR). We hypothesized that similar cells might also occur in peripheral blood. In 13/13 normal individuals, we found a circulating population of CD56(low), CD33+, FcgammaRI+, FcgammaRII+, HLA-DR+, CD11b(high), CD14+ monocytes closely resembling the cultured CD56(low)CD33+ cells. They may represent a normal counterpart of the CD56+ CD33+ hybrid myeloid/natural killer cell leukemia. Their mean frequency was 1.3+/-1% (standard deviation), range 0.16-3.5%, of total mononuclear cells. CD56(low)CD33+ cells, primed with cytomegalovirus antigen, induced autologous T-lymphocyte proliferation comparably to CD56-, CD14+ peripheral blood monocytes (PBM). Conversely, CD56(low) cells induced greater T cell proliferation than CD56- PBM when lymphocyte responders were HLA mismatched. Unstimulated CD56(low)CD33+ cells showed a low antiproliferative effect on K562, which was increased upon LPS stimulation. The pattern of cytokine production by CD56(low)CD33+ cells and PBM largely overlapped; however, they produced detectable levels of IL-6 and IL-1beta. These results define a minor monocyte population with distinct phenotypic and functional features. PMID- 15526028 TI - Is physical fitness decreased in survivors of childhood leukemia? A systematic review. AB - The aim of this review is to determine whether physical fitness, assessed by peak oxygen uptake (VO(2peak)) measurement, is reduced in survivors of acute lymphoblastic leukemia (ALL) compared to healthy children. A systematic literature search (up to June 2004) was performed using Medline, Sportdiscus, Cinahl, Embase, Cochrane and PEDro database and reference tracking. The VO(2peak) (ml kg(-1) min(-1)) reached during a maximal exercise test until volitional exhaustion was used as the main outcome for this review. In all, 17 studies were identified in the literature. Data from three studies (102 ALL survivors, age ranging from 7 to 19 years) were pooled in a meta-analysis. Although there was a significant heterogeneity between the included studies (P=0.0006), the standardized mean difference (SMD) value of -0.61 (P=0.07) indicated that VO(2peak) tended to be reduced in survivors of childhood ALL compared to healthy control subjects, that is, decrease of -5.97 ml kg(-1) min(-1) (95% confidence interval (CI): (-12.35, 0.41); P=0.07) or -13% (95 % CI: (-27, 0.004)). Physical fitness tends to be reduced in survivors of ALL during childhood, which suggests the need for this population group to engage in regular physical activities with the purpose of increasing their functional capacity. Although more research is needed, this functional improvement might ameliorate the quality of life of ALL survivors as physical and outdoors activities are an essential part of daily routine during childhood. PMID- 15526029 TI - Translocation t(1;19) is related to low cellular drug resistance in childhood acute lymphoblastic leukaemia. PMID- 15526030 TI - Axin stimulates p53 functions by activation of HIPK2 kinase through multimeric complex formation. AB - Axin and p53 are tumor suppressors, controlling cell growth, apoptosis, and development. We show that Axin interacts with homeodomain-interacting protein kinase-2 (HIPK2), which is linked to UV-induced p53-dependent apoptosis by interacting with, and phosphorylating Ser 46 of, p53. In addition to association with p53 via HIPK2, Axin contains a separate domain that directly interacts with p53 at their physiological concentrations. Axin stimulates p53-dependent reporter transcription in 293 cells, but not in 293T, H1299, or SaOS-2 cells that are defective in p53 signaling. Axin, but not AxindeltaHIPK2, activates HIPK2 mediated p53 phosphorylation at Ser 46, facilitating p53-dependent transcriptional activity and apoptosis. Specific knockdown of Axin by siRNA reduced UV-induced Ser-46 phosphorylation and apoptosis. Kinase-dead HIPK2 reduced Axin-induced p53-dependent transcriptional activity, indicating that Axin stimulates p53 function through HIPK2 kinase activity. Interestingly, HIPK2deltaAxin that lacks its Axin-binding region acts as a dominant-positive form in p53 activation, suggesting that the Axin-binding region of HIPK2 is a putative autoinhibitory domain. These results show that Axin acts as a tumor suppressor by facilitating p53 function through integration of multiple factors. PMID- 15526031 TI - Post-transfer editing in vitro and in vivo by the beta subunit of phenylalanyl tRNA synthetase. AB - Translation of the genetic code requires attachment of tRNAs to their cognate amino acids. Errors during amino-acid activation and tRNA esterification are corrected by aminoacyl-tRNA synthetase-catalyzed editing reactions, as extensively described for aliphatic amino acids. The contribution of editing to aromatic amino-acid discrimination is less well understood. We show that phenylalanyl-tRNA synthetase misactivates tyrosine and that it subsequently corrects such errors through hydrolysis of tyrosyl-adenylate and Tyr-tRNA(Phe). Structural modeling combined with an in vivo genetic screen identified the editing site in the B3/B4 domain of the beta subunit, 40 angstroms from the active site in the alpha subunit. Replacements of residues within the editing site had no effect on Phe-tRNA(Phe) synthesis, but abolished hydrolysis of Tyr tRNA(Phe) in vitro. Expression of the corresponding mutants in Escherichia coli significantly slowed growth, and changed the activity of a recoded beta galactosidase variant by misincorporating tyrosine in place of phenylalanine. This loss in aromatic amino-acid discrimination in vivo revealed that editing by phenylalanyl-tRNA synthetase is essential for faithful translation of the genetic code. PMID- 15526032 TI - Crosstalk between PKCzeta and the IL4/Stat6 pathway during T-cell-mediated hepatitis. AB - PKCzeta is required for nuclear factor kappa-B (NF-kappaB) activation in several cell systems. NF-kappaB is a suppressor of liver apoptosis during development and in concanavalin A (ConA)-induced T-cell-mediated hepatitis. Here we show that PKCzeta-/- mice display inhibited ConA-induced NF-kappaB activation and reduced damage in liver. As the IL-4/Stat6 pathway is necessary for ConA-induced hepatitis, we addressed here the potential role of PKCzeta in this cascade. Interestingly, the loss of PKCzeta severely attenuated serum IL-5 and liver eotaxin-1 levels, two critical mediators of liver damage. Stat6 tyrosine phosphorylation and Jak1 activation were ablated in the liver of ConA-injected PKCzeta-/- mice and in IL-4-stimulated PKCzeta-/- fibroblasts. PKCzeta interacts with and phosphorylates Jak1 and PKCzeta activity is required for Jak1 function. In contrast, Par-4-/- mice have increased sensitivity to ConA-induced liver damage and IL-4 signaling. This unveils a novel and critical involvement of PKCzeta in the IL-4/Stat6 signaling pathway in vitro and in vivo. PMID- 15526033 TI - The mitochondrial RNA polymerase contributes critically to promoter specificity in mammalian cells. AB - Initiation of transcription in mammalian mitochondria depends on three proteins: mitochondrial RNA polymerase (POLRMT), mitochondrial transcription factor A (TFAM) and mitochondrial transcription factor B2 (TFB2M). We show here that the recombinant mouse and human transcription machineries are unable to initiate transcription in vitro from the heterologous light-strand promoter (LSP) of mitochondrial DNA. This species specificity is dependent on the interaction of TFAM and POLRMT with specific distal and proximal promoter elements. A sequence element localized from position -1 to -2 relative to the transcription start site in LSP functionally interacts with POLRMT. The POLRMT/TFB2M heterodimer is unable to interact with promoter elements and initiate even abortive transcription in the absence of TFAM. TFAM is thus an integral part of the mammalian transcription machinery, and we propose that TFAM induces a structural change of the promoter that is required for POLRMT-dependent promoter recognition. PMID- 15526034 TI - Protection from cytosolic prion protein toxicity by modulation of protein translocation. AB - Failure to promptly dispose of undesirable proteins is associated with numerous diseases. In the case of cellular prion protein (PrP), inhibition of the proteasome pathway can generate a highly aggregation-prone, cytotoxic form of PrP implicated in neurodegeneration. However, the predominant mechanisms that result in delivery of PrP, ordinarily targeted to the secretory pathway, to cytosolic proteasomes have been unclear. By accurately measuring the in vivo fidelity of protein translocation into the endoplasmic reticulum (ER), we reveal a slight inefficiency in PrP signal sequence function that generates proteasomally degraded cytosolic PrP. Attenuating this source of cytosolic PrP completely eliminates the dependence on proteasomes for PrP degradation. This allows cells to tolerate both higher expression levels and decreased proteasomal capacity without succumbing to the adverse consequences of misfolded PrP. Thus, the generation of potentially toxic cytosolic PrP is controlled primarily during its initial translocation into the ER. These results suggest that a substantial proportion of the cell's constitutive proteasomal burden may consist of proteins that, like PrP, fail to cotranslationally enter the secretory pathway with high fidelity. PMID- 15526035 TI - AIF deficiency compromises oxidative phosphorylation. AB - Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein that, after apoptosis induction, translocates to the nucleus where it participates in apoptotic chromatinolysis. Here, we show that human or mouse cells lacking AIF as a result of homologous recombination or small interfering RNA exhibit high lactate production and enhanced dependency on glycolytic ATP generation, due to severe reduction of respiratory chain complex I activity. Although AIF itself is not a part of complex I, AIF-deficient cells exhibit a reduced content of complex I and of its components, pointing to a role of AIF in the biogenesis and/or maintenance of this polyprotein complex. Harlequin mice with reduced AIF expression due to a retroviral insertion into the AIF gene also manifest a reduced oxidative phosphorylation (OXPHOS) in the retina and in the brain, correlating with reduced expression of complex I subunits, retinal degeneration, and neuronal defects. Altogether, these data point to a role of AIF in OXPHOS and emphasize the dual role of AIF in life and death. PMID- 15526036 TI - Chemical validation of GPI biosynthesis as a drug target against African sleeping sickness. AB - It has been suggested that compounds affecting glycosylphosphatidylinositol (GPI) biosynthesis in bloodstream form Trypanosoma brucei should be trypanocidal. We describe cell-permeable analogues of a GPI intermediate that are toxic to this parasite but not to human cells. These analogues are metabolized by the T. brucei GPI pathway, but not by the human pathway. Closely related nonmetabolizable analogues have no trypanocidal activity. This represents the first direct chemical validation of the GPI biosynthetic pathway as a drug target against African human sleeping sickness. The results should stimulate further inhibitor design and synthesis and encourage the search for inhibitors in natural product and synthetic compound libraries. PMID- 15526037 TI - Functional interaction between p90Rsk2 and Emi1 contributes to the metaphase arrest of mouse oocytes. AB - Vertebrate eggs arrest at metaphase of the second meiotic division before fertilization under the effect of a cytostatic factor (CSF). This arrest is established during oocyte maturation by the MAPK kinase module, comprised of Mos, MEK, MAPKs and p90Rsk. Maintenance of CSF arrest at metaphase requires inhibitors of the anaphase-promoting complex (APC) like Emi1, which sequesters the APC activator Cdc20. Although it was proposed that the Mos pathway and Emi1 act independently, neither one alone is sufficient to entirely reproduce CSF arrest. Herein we demonstrate that p90Rsk2 associates with and phosphorylates Emi1 upstream of the binding region for Cdc20, thus stabilizing their interaction. Experiments in transfected cells and two-cell embryos indicate that Emi1 and p90Rsk2 cooperate to induce the metaphase arrest. Moreover, oocyte maturation was impaired by interfering with the interaction between p90Rsk2 and Emi1 or by RNA interference of Emi1. Our results indicate that p90Rsk2 and Emi1 functionally interact during oocyte maturation and that the Mos pathway establishes CSF activity through stabilization of an APC-inhibitory complex composed by Emi1 and Cdc20 before fertilization. PMID- 15526038 TI - The synaptic acetylcholinesterase tetramer assembles around a polyproline II helix. AB - Functional localization of acetylcholinesterase (AChE) in vertebrate muscle and brain depends on interaction of the tryptophan amphiphilic tetramerization (WAT) sequence, at the C-terminus of its major splice variant (T), with a proline-rich attachment domain (PRAD), of the anchoring proteins, collagenous (ColQ) and proline-rich membrane anchor. The crystal structure of the WAT/PRAD complex reveals a novel supercoil structure in which four parallel WAT chains form a left handed superhelix around an antiparallel left-handed PRAD helix resembling polyproline II. The WAT coiled coils possess a WWW motif making repetitive hydrophobic stacking and hydrogen-bond interactions with the PRAD. The WAT chains are related by an approximately 4-fold screw axis around the PRAD. Each WAT makes similar but unique interactions, consistent with an asymmetric pattern of disulfide linkages between the AChE tetramer subunits and ColQ. The P59Q mutation in ColQ, which causes congenital endplate AChE deficiency, and is located within the PRAD, disrupts crucial WAT-WAT and WAT-PRAD interactions. A model is proposed for the synaptic AChE(T) tetramer. PMID- 15526040 TI - Steps on the critical path: arresting HIV/AIDS in developing countries. PMID- 15526039 TI - The molecular basis for oat intolerance in patients with celiac disease. AB - BACKGROUND: Celiac disease is a small intestinal inflammatory disorder characterized by malabsorption, nutrient deficiency, and a range of clinical manifestations. It is caused by an inappropriate immune response to dietary gluten and is treated with a gluten-free diet. Recent feeding studies have indicated oats to be safe for celiac disease patients, and oats are now often included in the celiac disease diet. This study aimed to investigate whether oat intolerance exists in celiac disease and to characterize the cells and processes underlying this intolerance. METHODS AND FINDINGS: We selected for study nine adults with celiac disease who had a history of oats exposure. Four of the patients had clinical symptoms on an oats-containing diet, and three of these four patients had intestinal inflammation typical of celiac disease at the time of oats exposure. We established oats-avenin-specific and -reactive intestinal T cell lines from these three patients, as well as from two other patients who appeared to tolerate oats. The avenin-reactive T-cell lines recognized avenin peptides in the context of HLA-DQ2. These peptides have sequences rich in proline and glutamine residues closely resembling wheat gluten epitopes. Deamidation (glutamine-->glutamic acid conversion) by tissue transglutaminase was involved in the avenin epitope formation. CONCLUSIONS: We conclude that some celiac disease patients have avenin-reactive mucosal T-cells that can cause mucosal inflammation. Oat intolerance may be a reason for villous atrophy and inflammation in patients with celiac disease who are eating oats but otherwise are adhering to a strict gluten-free diet. Clinical follow-up of celiac disease patients eating oats is advisable. PMID- 15526041 TI - The future of surgical research. PMID- 15526042 TI - How should the health community respond to violent political conflict? PMID- 15526043 TI - Generalized seizure in a Mauritian woman taking bupropion. PMID- 15526044 TI - Assessment of volume depletion in children with malaria. AB - BACKGROUND: The degree of volume depletion in severe malaria is currently unknown, although knowledge of fluid compartment volumes can guide therapy. To assist management of severely ill children, and to test the hypothesis that volume changes in fluid compartments reflect disease severity, we measured body compartment volumes in Gabonese children with malaria. METHODS AND FINDINGS: Total body water volume (TBW) and extracellular water volume (ECW) were estimated in children with severe or moderate malaria and in convalescence by tracer dilution with heavy water and bromide, respectively. Intracellular water volume (ICW) was derived from these parameters. Bioelectrical impedance analysis estimates of TBW and ECW were calibrated against dilution methods, and bioelectrical impedance analysis measurements were taken daily until discharge. Sixteen children had severe and 19 moderate malaria. Severe childhood malaria was associated with depletion of TBW (mean [SD] of 37 [33] ml/kg, or 6.7% [6.0%]) relative to measurement at discharge. This is defined as mild dehydration in other conditions. ECW measurements were normal on admission in children with severe malaria and did not rise in the first few days of admission. Volumes in different compartments (TBW, ECW, and ICW) were not related to hyperlactataemia or other clinical and laboratory markers of disease severity. Moderate malaria was not associated with a depletion of TBW. CONCLUSIONS: Significant hypovolaemia does not exacerbate complications of severe or moderate malaria. As rapid rehydration of children with malaria may have risks, we suggest that fluid replacement regimens should aim to correct fluid losses over 12-24 h. PMID- 15526045 TI - Nevirapine and efavirenz elicit different changes in lipid profiles in antiretroviral-therapy-naive patients infected with HIV-1. AB - BACKGROUND: Patients infected with HIV-1 initiating antiretroviral therapy (ART) containing a non-nucleoside reverse transcriptase inhibitor (NNRTI) show presumably fewer atherogenic lipid changes than those initiating most ARTs containing a protease inhibitor. We analysed whether lipid changes differed between the two most commonly used NNRTIs, nevirapine (NVP) and efavirenz (EFV). METHODS AND FINDINGS: Prospective analysis of lipids and lipoproteins was performed in patients enrolled in the NVP and EFV treatment groups of the 2NN study who remained on allocated treatment during 48 wk of follow-up. Patients were allocated to NVP (n = 417), or EFV (n = 289) in combination with stavudine and lamivudine. The primary endpoint was percentage change over 48 wk in high density lipoprotein cholesterol (HDL-c), total cholesterol (TC), TC:HDL-c ratio, non-HDL-c, low-density lipoprotein cholesterol, and triglycerides. The increase of HDL-c was significantly larger for patients receiving NVP (42.5%) than for patients receiving EFV (33.7%; p = 0.036), while the increase in TC was lower (26.9% and 31.1%, respectively; p = 0.073), resulting in a decrease of the TC:HDL c ratio for patients receiving NVP (-4.1%) and an increase for patients receiving EFV (+5.9%; p < 0.001). The increase of non-HDL-c was smaller for patients receiving NVP (24.7%) than for patients receiving EFV (33.6%; p = 0.007), as were the increases of triglycerides (20.1% and 49.0%, respectively; p < 0.001) and low density lipoprotein cholesterol (35.0% and 40.0%, respectively; p = 0.378). These differences remained, or even increased, after adjusting for changes in HIV-1 RNA and CD4+ cell levels, indicating an effect of the drugs on lipids over and above that which may be explained by suppression of HIV-1 infection. The increases in HDL-c were of the same order of magnitude as those seen with the use of the investigational HDL-c-increasing drugs. CONCLUSION: NVP-containing ART shows larger increases in HDL-c and decreases in TC:HDL-c ratio than an EFV-containing regimen. Based on these findings, protease-inhibitor-sparing regimens based on non-nucleoside reverse transcriptase inhibitor, particularly those containing NVP, may be expected to result in a reduced risk of coronary heart disease. PMID- 15526047 TI - Characterization of T lymphocytes in chronic obstructive pulmonary disease. PMID- 15526049 TI - Distribution of major health risks: findings from the Global Burden of Disease study. AB - BACKGROUND: Most analyses of risks to health focus on the total burden of their aggregate effects. The distribution of risk-factor-attributable disease burden, for example by age or exposure level, can inform the selection and targeting of specific interventions and programs, and increase cost-effectiveness. METHODS AND FINDINGS: For 26 selected risk factors, expert working groups conducted comprehensive reviews of data on risk-factor exposure and hazard for 14 epidemiological subregions of the world, by age and sex. Age-sex-subregion population attributable fractions were estimated and applied to the mortality and burden of disease estimates from the World Health Organization Global Burden of Disease database. Where possible, exposure levels were assessed as continuous measures, or as multiple categories. The proportion of risk-factor-attributable burden in different population subgroups, defined by age, sex, and exposure level, was estimated. For major cardiovascular risk factors (blood pressure, cholesterol, tobacco use, fruit and vegetable intake, body mass index, and physical inactivity) 43%-61% of attributable disease burden occurred between the ages of 15 and 59 y, and 87% of alcohol-attributable burden occurred in this age group. Most of the disease burden for continuous risks occurred in those with only moderately raised levels, not among those with levels above commonly used cut-points, such as those with hypertension or obesity. Of all disease burden attributable to being underweight during childhood, 55% occurred among children 1 3 standard deviations below the reference population median, and the remainder occurred among severely malnourished children, who were three or more standard deviations below median. CONCLUSIONS: Many major global risks are widely spread in a population, rather than restricted to a minority. Population-based strategies that seek to shift the whole distribution of risk factors often have the potential to produce substantial reductions in disease burden. PMID- 15526050 TI - The global health watch. PMID- 15526053 TI - Palliative care in Africa and the Caribbean. PMID- 15526052 TI - Should health professionals screen all women for domestic violence? AB - BACKGROUND TO THE DEBATE: The US and Canadian task forces on preventive health recently declared that there is not enough evidence to recommend for or against routine universal screening of women for domestic violence. Yet some experts argue that routine enquiry is justified. PMID- 15526054 TI - New drugs for neglected diseases: from pipeline to patients. PMID- 15526055 TI - Fever, headache, and visual blurring in a 17-year-old woman. PMID- 15526057 TI - The birth of reproductive health: a difficult delivery. PMID- 15526056 TI - An immune basis for lung parenchymal destruction in chronic obstructive pulmonary disease and emphysema. AB - BACKGROUND: Chronic obstructive pulmonary disease and emphysema are a frequent result of long-term smoking, but the exact mechanisms, specifically which types of cells are associated with the lung destruction, are unclear. METHODS AND FINDINGS: We studied different subsets of lymphocytes taken from portions of human lungs removed surgically to find out which lymphocytes were the most frequent, which cell-surface markers these lymphocytes expressed, and whether the lymphocytes secreted any specific factors that could be associated with disease. We found that loss of lung function in patients with chronic obstructive pulmonary disease and emphysema was associated with a high percentage of CD4+ and CD8+ T lymphocytes that expressed chemokine receptors CCR5 and CXCR3 (both markers of T helper 1 cells), but not CCR3 or CCR4 (markers of T helper 2 cells). Lung lymphocytes in patients with chronic obstructive pulmonary disease and emphysema secrete more interferon gamma--often associated with T helper 1 cells- and interferon-inducible protein 10 and monokine induced by interferon, both of which bind to CXCR3 and are involved in attracting T helper 1 cells. In response to interferon-inducible protein 10 and monokine induced by interferon, but not interferon gamma, lung macrophages secreted macrophage metalloelastase (matrix metalloproteinase-12), a potent elastin-degrading enzyme that causes tissue destruction and which has been linked to emphysema. CONCLUSIONS: These data suggest that Th1 lymphoctytes in the lungs of people with smoking-related damage drive progression of emphysema through CXCR3 ligands, interferon-inducible protein 10, and monokine induced by interferon. PMID- 15526060 TI - Video views and reviews: Golgi export, targeting, and plasma membrane caveolae. PMID- 15526058 TI - A randomised, double-blind, controlled vaccine efficacy trial of DNA/MVA ME-TRAP against malaria infection in Gambian adults. AB - BACKGROUND: Many malaria vaccines are currently in development, although very few have been evaluated for efficacy in the field. Plasmodium falciparum multiple epitope (ME)- thrombospondin-related adhesion protein (TRAP) candidate vaccines are designed to potently induce effector T cells and so are a departure from earlier malaria vaccines evaluated in the field in terms of their mechanism of action. ME-TRAP vaccines encode a polyepitope string and the TRAP sporozoite antigen. Two vaccine vectors encoding ME-TRAP, plasmid DNA and modified vaccinia virus Ankara (MVA), when used sequentially in a prime-boost immunisation regime, induce high frequencies of effector T cells and partial protection, manifest as delay in time to parasitaemia, in a clinical challenge model. METHODS AND FINDINGS: A total of 372 Gambian men aged 15-45 y were randomised to receive either DNA ME-TRAP followed by MVA ME-TRAP or rabies vaccine (control). Of these men, 296 received three doses of vaccine timed to coincide with the beginning of the transmission season (141 in the DNA/MVA group and 155 in the rabies group) and were followed up. Volunteers were given sulphadoxine/pyrimethamine 2 wk before the final vaccination. Blood smears were collected weekly for 11 wk and whenever a volunteer developed symptoms compatible with malaria during the transmission season. The primary endpoint was time to first infection with asexual P. falciparum. Analysis was per protocol. DNA ME-TRAP and MVA ME-TRAP were safe and well-tolerated. Effector T cell responses to a non-vaccine strain of TRAP were 50-fold higher postvaccination in the malaria vaccine group than in the rabies vaccine group. Vaccine efficacy, adjusted for confounding factors, was 10.3% (95% confidence interval, -22% to +34%; p = 0.49). Incidence of malaria infection decreased with increasing age and was associated with ethnicity. CONCLUSIONS: DNA/MVA heterologous prime-boost vaccination is safe and highly immunogenic for effector T cell induction in a malaria-endemic area. But despite having produced a substantial reduction in liver-stage parasites in challenge studies of non-immune volunteers, this first generation T cell-inducing vaccine was ineffective at reducing the natural infection rate in semi-immune African adults. PMID- 15526061 TI - The 50th Anniversary of Brown v. Board of Education: continued impacts on minority life science education. PMID- 15526059 TI - Limited durability of viral control following treated acute HIV infection. AB - BACKGROUND: Early treatment of acute HIV infection with highly active antiretroviral therapy, followed by supervised treatment interruption (STI), has been associated with at least transient control of viremia. However, the durability of such control remains unclear. Here we present longitudinal follow up of a single-arm, open-label study assessing the impact of STI in the setting of acute HIV-1 infection. METHODS AND FINDINGS: Fourteen patients were treated during acute HIV-1 infection and subsequently subjected to an STI protocol that required retreatment if viral load exceeded 50,000 RNA copies/ml plasma or remained above 5,000 copies/ml for more than three consecutive weeks. Eleven of 14 (79%) patients were able to achieve viral loads of less than 5,000 RNA copies/ml for at least 90 d following one, two, or three interruptions of treatment. However, a gradual increase in viremia and decline in CD4+ T cell counts was observed in most individuals. By an intention-to-treat analysis, eight (57%), six (43%), and three (21%) of 14 patients achieved a maximal period of control of 180, 360, and 720 d, respectively, despite augmentation of HIV specific CD4+ and CD8+ T cell responses. The magnitude of HIV-1-specific cellular immune responses before treatment interruption did not predict duration of viremia control. The small sample size and lack of concurrent untreated controls preclude assessment of possible clinical benefit despite failure to control viremia by study criteria. CONCLUSIONS: These data indicate that despite initial control of viremia, durable viral control to less than 5,000 RNA copies/ml plasma in patients following treated acute HIV-1 infection occurs infrequently. Determination of whether early treatment leads to overall clinical benefit will require a larger and randomized clinical trial. These data may be relevant to current efforts to develop an HIV-1 vaccine designed to retard disease progression rather than prevent infection since they indicate that durable maintenance of low-level viremia may be difficult to achieve. PMID- 15526062 TI - Outcomes from the undergraduate research summit. PMID- 15526063 TI - PowerPoint in the classroom. AB - Points of View (POV) addresses issues faced within life science education. Cell Biology Education has launched the POV feature to present two or more opinions published in tandem on a common topic. We consider POVs to be "Op-Ed" pieces designed to stimulate thought and dialog on significant educational issues. Each author has the opportunity to revise a POV after reading drafts of the other POVs. In this issue, we ask the question, "Is PowerPoint the best instructional medium to use in your class?" Everyone seems to have an opinion on Microsoft, but the intellectual merits of using PowerPoint (or similar software) is a growing question as states and institutions put more and more money into information technology and distance learning. Four POVs are presented: 1) David Keefe and James Willett provide their case why PowerPoint is an ideal teaching software. Keefe is an educational researcher at the Center for Technology in Learning at SRI International. Willett is a professor at George Mason University in the Departments of Microbial and Molecular Bioscience; as well as Bioinformatics and Computational Biology. 2) Kim McDonald highlights the causes of PowerPointlessness, a term which indicates the frequent use of PowerPoint as a crutch rather than a tool. She is a Bioscience Educator at the Shodor Education Foundation, Inc. 3) Diana Voss asks readers if PowerPoint is really necessary to present the material effectively or not. Voss is a Instructional Computing Support Specialist at SUNY Stony Brook. 4) Cynthia Lanius takes a light-hearted approach to ask whether PowerPoint is a technological improvement or just a change of pace for teacher and student presentations. Lanius is a Technology Integration Specialist in the Sinton (Texas) Independent School District. The authors span the range of teaching experiences and settings from which they bring different points of view to the debate. Readers are encouraged to participate in the online discussion forum hosted by CBE at www.cellbioed.org/discussion/public/main.cfm and/or contact the authors directly. PMID- 15526064 TI - The University of Alabama at Birmingham Center for Community OutReach Development Summer Science Institute Program: a 3-yr laboratory research experience for inner city secondary-level students. AB - This article describes and assesses the effectiveness of a 3-yr, laboratory-based summer science program to improve the academic performance of inner-city high school students. The program was designed to gradually introduce such students to increasingly more rigorous laboratory experiences in an attempt to interest them in and model what "real" science is like. The students are also exposed to scientific seminars and university tours as well as English and mathematics workshops designed to help them analyze their laboratory data and prepare for their closing ceremony presentations. Qualitative and quantitative analysis of student performance in these programs indicates that participants not only learn the vocabulary, facts, and concepts of science, but also develop a better appreciation of what it is like to be a "real" scientist. In addition, the college-bound 3-yr graduates of this program appear to be better prepared to successfully academically compete with graduates of other high schools; they also report learning useful job-related life skills. Finally, the critical conceptual components of this program are discussed so that science educators interested in using this model can modify it to fit the individual resources and strengths of their particular setting. PMID- 15526065 TI - Use of animation in teaching cell biology. AB - To address the different learning styles of students, and because students can access animation from off-campus computers, the use of digital animation in teaching cell biology has become increasingly popular. Sample processes from cell biology that are more clearly presented in animation than in static illustrations are identified. The value of animation is evaluated on whether the process being taught involves motion, cellular location, or sequential order of numerous events. Computer programs for developing animation and animations associated with cell biology textbooks are reviewed, and links to specific examples of animation are given. Finally, future teaching tools for all fields of biology will increasingly benefit from an expansion of animation to the use of simulation. One purpose of this review is to encourage the widespread use of animations in biology teaching by discussing the nature of digital animation. PMID- 15526066 TI - Assessing precollege science education outreach initiatives: a funder's perspective. AB - We describe an assessment of the collective impact of 35 grants that the Howard Hughes Medical Institute (HHMI) made to biomedical research institutions in 1999 to support precollege science education outreach programs. Data collected from funded institutions were compared with data from a control group of institutions that had advanced to the last stage of review but had not been funded. The survey instrument and the results reveal outcomes and impacts that HHMI considers relevant for these programs. The following attributes are considered: ability to secure additional, non-HHMI funding; institution buy-in as measured by gains in dedicated space and staff; enhancement of the program director's career; number and adoption of educational products developed; number of related publications and awards; percentage of programs for which teachers received course credit; increase in science content knowledge; and increase in student motivation to study science. PMID- 15526067 TI - [Congenital deficiencies of coagulation factors and acquired inhibitors leading to bleeding disorders]. AB - This review focuses on symptoms, course and treatment of bleeding disorders due to hereditary coagulation factor deficiencies and acquired inhibitors, mentioning as well the pathophysiologic and molecular genetic aspects and diagnostic particularities. The review of haemophilia A and B deals with carrier problems, replacement therapy, additional haemostatic agents such as antifibrinolytics and desmopressin, the treatment of typical haemorrhages, haemophilia with inhibitors and future therapeutic options. Of the autosomal homozygous bleeding disorders such as von Willebrand disease type 3, afibrinogenaemia, factor XIII-, VII- and XI-deficiency each has its particularities influencing treatment strategies. The last chapter discusses acquired bleeding disorders such as acquired haemophilia A, von Willebrand disease, factor V deficiency and the hypoprothrombinaemia lupus anticoagulant syndrome, the different modes of inhibition, diagnostics and principles of treatment. PMID- 15526068 TI - [Thrombophilia]. AB - A survey on definitions, epidemiology, clinical manifestations of congenital thrombophilias and clinical relevant acquired thrombophilias is given. Diagnostic and therapeutical strategies are presented. PMID- 15526069 TI - [Acquired blood coagulation disorders]. AB - Acquired coagulopathies regularly result from underlying diseases such as liver failure or sepsis. Optimal treatment of these underlying diseases defines longterm prognosis. In a few coagulopathies, such as immunocoagulopathies, in a relevant portion of patients an underlying disease cannot be identified. Therapeutic modulation of haemostasis aiming at the prevention of fatal bleeding or thromboembolism is able to improve the prognosis of patients with acquired coagulopathies. Most often this is the result of individualized, lab-controlled treatments, which are difficult to be studied. Within the last years the role of therapeutic modulation of haemostasis to reduce mortality was successfully studied for the first time in the frame of randomized placebo-controlled phase III studies. PMID- 15526070 TI - [Hemostasis disorders in obstetrics and gynecology]. AB - A survey is given on the clinical important haemostatic facts and diseases in gynaecology and obstetrics from the viewpoint of the gynecologist and obstetrician. PMID- 15526071 TI - [Heparin, thrombin and Factor Xa inhibitors]. AB - Direct and indirect coagulation inhibitors are used to inhibit the activity of the serine proteases of the coagulation system. Indirect inhibitors act via antithrombin and heparin cofactor II. The main representatives are heparins, lowmolecular-weight heparins, fondaparinux, idraparinux and danaparoid. They bind to antithrombin and potentiate the inactivation of factor Xa and other serine proteases. Direct thrombin inhibitors bind reversibly to thrombin without cofactor. Anticoagulants are determined by global and specific anticoagulant methods. New anticoagulants are developed such as oral factor Xa inhibitors, oral thrombin inhibitors, antibody against activated factor VII, recombinant tissue pathway inhibitor to improve inhibition of blood coagulation or to induce nonanticoagulant effects (e. g. activated protein C in septicaemia). New anticoagulant methods are developed to improve and specify the anticoagulant effect of anticoagulants in thromboembolic diseases. PMID- 15526072 TI - [Oral anticoagulants and coumarin classes]. AB - A survey is given on pharmacology and indications for the treatment with vitamin K antagonists. The therapeutic handling and self control by the patient is described. PMID- 15526073 TI - [Clotting factor concentrates]. AB - Topics of this survey are the indications and use of plasma-derived and recombinant coagulation factor concentrates. These substitution therapies need to be handled carefully by weighing up their effectiveness against the long and short term side effects in the individual patient. PMID- 15526074 TI - Clinical holistic medicine: developing from asthma, allergy, and eczema. AB - This paper shows how consciousness-based holistic medicine can be used in the case of asthma, allergy, and eczema. We have many fine drugs to relieve patients from the worst of these symptoms, where many children and adults suffer health problems related to hyper-reactivity of the immune system. Many symptoms remain throughout life because the drugs do not cure the allergy and allergy today is the sixth leading cause of chronic illness. The etiology of the immune disturbances is mostly unknown from a biomedical perspective. Consciousness-based holistic medicine could therefore be used to treat these diseases if the patient is willing to confront hidden existential pain, is motivated to work hard, and is dedicated to improve quality of life, quality of working life, and personal relationships. Improving quality of life is not always an easy job for the patient, but it can be done with coaching from the physician. An increased physical health is often observed after only a few sessions with a physician skilled in using holistic medical tools and able to coach the patient successfully through a few weeks of dedicated homework. Children with allergy and asthma can also be helped if their parents are able to do work on personal development, to improve the general quality of life in the family and their relationship with the child. PMID- 15526075 TI - [Influence of environmental polluting ozone on the the skin]. AB - The skin is directly and frequently exposed to a pro-oxidative environment, including ozone and UV-radiation. While ozone in the stratosphere protects against mutagenic UVC-radiation, it is also a major air pollutant in urban areas. With its strong oxidizing potential, ozone is perhaps one of the most reactive chemicals the skin ever encounters. Although a large body of evidence exists for ozone- induced oxidative stress in the respiratory tract, the current knowledge on its in vivo effect on cutaneous tissues is based on studies of the last 10 years. Acute ozone exposure damages the stratum corneum, depletes skin vitamin C and E and induces lipid and protein oxidation in upper epidermal layers. Secondary products penetrate into deeper skin layers and are capable of activating signal transduction pathways and inducing cell damage. It has been shown in a murine model, that environmentally relevant ozone concentrations can induce a stress response in the skin. PMID- 15526076 TI - [Biomaterial studies in cultures of human stapedial bone-like cells]. AB - BACKGROUND: Cell culture studies may provide information on the behavior of biomaterials in the intended implant environment. Cell cultures from such an environment could be used for the development of middle ear implants. MATERIAL AND METHODS: Secondary bone-like cell cultures derived from human stapes were exposed to different materials [Al(2)O(3) ceramic, glass ceramic (Ceravital), gold and titanium]. Proliferation was studied for up to 40 days. RESULTS: The proliferation of cultured stapes bone-like cells did not differ significantly between the four tested biomaterials. The well known cytotoxic effect of copper, which was used as a control, was evident. CONCLUSIONS: Four biomaterials [Al(2)O(3) ceramic, glass ceramic (Ceravital), gold and titanium] have similar biocompatibility and no toxicity when tested in human stapes cell cultures. This in vitro model may be of considerable value for the further development of middle ear implants, e.g., when coated with bone morphogenetic proteins. PMID- 15526077 TI - [Intraoperative endosonographic guided resection of tongue carcinoma]. AB - BACKGROUND: Exact estimation of a tumor's size and the definition of adequate resection margins in carcinomas of the tongue are often difficult because of the tumor's extension and deep infiltration. METHODS: We have developed a method that allows intraoperative visualisation and marking of tumor margins. Intra-operative endosonography was performed on nine patients with carcinomas of the tongue using a 8-12 MHz linear array transducer. The oral cavity was flooded with normal saline solution and the transducer was immersed therein. This allowed scanning in a non-contact mode. The tumor margins were marked with a surgical suture under endosonographic monitoring. RESULTS: In the nine patients studied, the histological margins corresponded to the sonographic margins. The sonographic marking proved to be useful during the resection of the tumor and histological safety margins were respected in each case. CONCLUSIONS: This non-invasive procedure provides a quick and reliable orientation during the resection of tongue carcinoma, and a more precise and individual definition of resection margins is possible. Intraoperative non-contact use of endosonography is a promising method. PMID- 15526078 TI - [Differential therapy of heart failure. Which drug for which patient?]. AB - The implementation of guidelines for medical therapy of heart failure may be problematic for the following reasons: 1. Elderly patients and women were underrepresented in large clinical trials which may limit their therapeutic impact in these patients. 2. Therapeutic decisions are influenced by co morbidities like renal failure, obstructive airway disease (COLD, Asthma), stroke, and diabetes mellitus. We therefore discuss the differential therapy of heart failure in view of particular patient subgroups. PMID- 15526079 TI - [Circumscribed acral hypokeratosis]. AB - Circumscribed acral hypokeratosis is a rare chronic disorder of cornification that occurs predominantly in women. Lesions are solitary and do not respond to any local conservative treatment. They have to be differentiated by biopsy from other non-healing lesions and tumors in acral skin. Clinically lesions appear as sharply circumscribed reddish macules. The histologic hallmark is a circumscribed loss of the entire stratum corneum, which can be best demonstrated at the border of the lesion as a contrast to the perilesional broad stratum corneum typical for acral sites. PMID- 15526080 TI - Functional analysis of a mutant form of the receptor tyrosine kinase Tie2 causing venous malformations. AB - Tie2 is expressed predominantly in endothelial cells and is required for blood vessel formation and maintenance. A missense mutation resulting in an R to W substitution in the kinase domain of Tie2 co-segregates with an autosomal dominantly inherited form of vascular dysmorphogenesis, venous malformation (VM). The mechanism by which this activating mutation leads to vessel dysmorphogenesis in VM is not known. Here we examined Tie2 activation status in VM and found activated receptor in lesional and non-lesional vessels. To gain insight into functional effects of VM mutant Tie2, wild-type and R849W mutant receptor were expressed in cultured human venous endothelial cells. Mutant Tie2 was constitutively phosphorylated in endothelial cells in vivo and caused a marked suppression of apoptosis. The anti-apoptotic kinase Akt was constitutively activated in cells expressing mutant receptor. Dominant-negative Akt inhibited the pro-survival activity of mutant Tie2. Migration of smooth muscle cells induced by conditioned medium from cells expressing mutant receptor was similar to that from cells expressing wild-type receptor. These data suggest that a primary effect of R849W Tie2 in VM is to allow survival of mural cell poor vessels via ligand-independent Tie2 activation of Akt and endothelial survival, rather than to directly induce formation of dysmorphogenic vessels. PMID- 15526081 TI - [Shoulder Arthroplasty. Surgical management]. AB - Shoulder arthroplasty covers an extensive indication spectrum of degenerative, inflammatory to infectious, tumorous, instability-associated and neurogenic disease to acute trauma and posttraumatic joint destruction. Each of these indications requires its own regime and an appropriate implant. Osteoarthritis is the most frequent indication, and will be used as an example for surgical management, bringing the manufacturer-nonspecific general guidelines, joint mechanics and surgical technology into agreement. The fundamentals of the technique of implantation refer to the anatomical adaptability of the third generation of shoulder prostheses, which permits an accurate reconstruction of glenohumeralen joint centring. This article also describes far-reaching principles, which are generally applicable to all indications for shoulder arthroplasty. PMID- 15526082 TI - [Combined Scanners (PET/CT, SPECT/CT) Versus Multimodality Imaging with Separated Systems]. AB - With increasing use of combined PET/CT scanners in the last few years, multimodality imaging (Nuclear Medicine/Radiology) found its way into clinical routine diagnostics. In this overview, necessary components for multimodality imaging, strategies for image analysis and image presentation, and diagnostic goals of combined imaging are demonstrated and discussed. A special focus is on the question, whether combined scanners can be replaced by a software approach with separated modalities. Advantages and limitations of multimodality imaging with combined or separated scanners are shown. PMID- 15526083 TI - [Randomized phase III study on nonseminomatous testicular tumor]. PMID- 15526084 TI - [Nephrectomy in acute uncomplicated pyelonephritis]. AB - BACKGROUND: Which complications can develop from uncomplicated pyelonephritis that may require nephrectomy? METHODS: From January 1999 to June 2003 we carried out nephrectomy in ten cases due to abscess formation after acute, uncomplicated pyelonephritis. The medical files were evaluated retrospectively. RESULTS: Nine women and one man were involved. The mean age was 36.2 years. Leading symptoms: flank pain, fever and chills. The mean symptom duration before admission was 14.6 days. Urinary tract infections were caused by Escherichia coli (six), E. coli and Enterococcus (once) and Klebsiella pneumoniae (once). Two cultures were sterile. Indications for nephrectomy were urosepsis (7 cases), anuria (once), increasing abscess formation under antibiotic therapy (once), drastic deterioration of general condition (once). CONCLUSIONS: Uncomplicated pyelonephritis is easy to treat under outpatient conditions with adequate oral antibiotic therapy. Close control must be ensured and clear recovery of symptoms should occur within 48 h. In the case of long duration of symptoms (>6-7 days) or lack of improvement of symptoms under calculated therapy in the first 2 days, inpatient therapy should be initiated because of the high risk of infectious complications. According to our experience, the following patient group is especially at risk: female, symptom duration of at least approximately 1 week, pre-treatment and transfer from another departments. PMID- 15526085 TI - On the origin of six-rowed barley with brittle rachis, agriocrithon [Hordeum vulgare ssp. vulgare f. agriocrithon (Aberg) Bowd.], based on a DNA marker closely linked to the vrs1 (six-row gene) locus. AB - The origin of six-rowed cultivated barley has been revealed to be more complex since the discovery of agriocrithon, a six-rowed barley with brittle rachis. The present study investigates whether such six-rowed brittle barley is wild or hybrid in nature, by analyzing genetic diversity at the cMWG699 marker locus, which is closely linked to the vrs1 (six-row gene) locus. DNA sequence analysis for 42 accessions showed only three types in six-rowed brittle barleys; in contrast, nine sequence types were found in ten wild barleys, ssp. spontaneum, in our previous study. Nucleotide diversities for the six-rowed brittle barley were 2.8-4.5 times lower than that for the ssp. spontaneum at this marker locus. The three sequence types found in the six-rowed brittle barley also appeared in the six-rowed cultivated barley. A cross-allelism test confirmed that the six-rowed character of the six-rowed brittle barley was controlled by the vrs1 locus. The nucleotide diversity and genealogy demonstrated that f. agriocrithon does not have the same level of diversity as found in wild barley, ssp. spontaneum. Consequently, f. agriocrithon does not appear to represent genuinely wild populations, but more probably originated from hybridization between ssp. spontaneum and six-rowed cultivated barley. PMID- 15526086 TI - Functionally associated molecular genetic marker map construction in perennial ryegrass (Lolium perenne L.). AB - A molecular marker-based map of perennial ryegrass (Lolium perenne L.) has been constructed through the use of polymorphisms associated with expressed sequence tags (ESTs). A pair-cross between genotypes from a North African ecotype and the cultivar Aurora was used to generate a two-way pseudo-testcross population. A selection of 157 cDNAs assigned to eight different functional categories associated with agronomically important biological processes was used to detect polymorphic EST-RFLP loci in the F(1)(NA(6) x AU(6)) population. A comprehensive set of EST-SSR markers was developed from the analysis of 14,767 unigenes, with 310 primer pairs showing efficient amplification and detecting 113 polymorphic loci. Two parental genetic maps were produced: the NA(6) genetic map contains 88 EST-RFLP and 71 EST-SSR loci with a total map length of 963 cM, while the AU(6) genetic map contains 67 EST-RFLP and 58 EST-SSR loci with a total map length of 757 cM. Bridging loci permitted the alignment of homologous chromosomes between the parental maps, and a sub-set of genomic DNA-derived SSRs was used to relate linkage groups to the perennial ryegrass reference map. Regions of segregation distortion were identified, in some instances in common with other perennial ryegrass maps. The EST-derived marker-based map provides the basis for in silico comparative genetic mapping, as well as the evaluation of co-location between QTLs and functionally associated genetic loci. PMID- 15526089 TI - Factors associated with mortality and functional disability after hip fracture: an inception cohort study. AB - Hip fracture results in excess mortality and functional disability. This study sought to identify predictors of mortality and limited functional ability 1 year after hip fracture. We conducted a 1-year follow-up of a prospective population based inception cohort of 218 hip fracture patients who had been consecutively admitted and discharged from hospital during the previous year. Mortality was observed to be independently associated with poor mental status (relative risk [RR]=6.96; 95% confidence interval [95% CI], 1.73-28.00), prefracture limited functional ability (RR=4.35; 95% CI, 1.32-14.36), institutionalized disposition at discharge (RR = 2.92; 95% CI, 1.02-8.38), and male gender (RR = 2.44; 95% CI, 1.01-5.93). Independent predictors of limited functional ability were prefracture functional disability (RR = 34.14; 95% CI, 3.13-372.33), poor mental status (RR = 9.71; 95% CI, 1.57-59.82), age >80 years (RR = 4.03; 95% CI, 1.48-11.00), and female gender (RR = 3.57; 95% CI, 0.08-0.98). On discharge, special attention and care should thus be given to all patients displaying any of the above predictive factors. PMID- 15526090 TI - Fractures of the proximal humerus in osteoporotic bone. AB - The purpose of this article was to review critically the current treatment options for fractures of the proximal humerus in patients with severe osteoporosis. The main difficulties lie in correctly diagnosing the fracture and hence selecting the most appropriate method of treatment. The reliability of the diagnosis can be increased by systematically appending additional information to a basic fracture classification. Classification is best carried out on a morphological basis, whereby a descriptor of bone quality can be added in order to introduce the degree of osteoporosis into the decision-making algorithm. Any classification system that claims to provide both treatment and prognosis is inappropriate, because prognosis will depend hopefully on the treatment. Approaches to treatment differ widely amongst centers and surgeons. It is still unclear as to what would be the optimal treatment. Factors such as the individual's functional requirements and ability to cooperate should be given careful consideration. At our institution, hemiarthroplasty is the method of choice for ischemic humeral heads and/or when anatomic reconstruction cannot be obtained. In all other displaced fractures, the main objective is preservation of the head since the best functional results can generally be obtained with internal fixation. Selection of a balanced osteosynthesis, adapted to the weak bone, is mandatory. Bulky, stiff implants are inadequate and may cause additional damage. Load sharing, not load bearing, compound constructions are the aim. Obtaining metaphyseal elastic buttressing is the key element in achieving the necessary load-sharing fixation. The system should allow controlled impaction and be forgiving towards occasional load peaks that will occur and are beyond patient control. Thin and flexible implants are required to realize this type of fixation. Given the polypragmatic approach that is current in clinical practice there is room for further improvement of techniques and implants. PMID- 15526091 TI - Renal effects and erythrocyte oxidative stress in long-term low-level lead exposed adolescent workers in auto repair workshops. AB - Lead poisoning is an old but persistent public health problem in developing countries. The present study investigated blood lead levels and its effects on markers of renal function and parameters of erythrocyte oxidative stress in adolescent male auto repair workers in Turkey. Blood Pb level and the ALAD index (logarithm of activated delta-aminolaevulinic acid dehydratase/nonactivated delta aminolaevulinic acid dehydratase) were measured as indicators of exposure to Pb. Markers of tubular damage urine N-acetyl-beta-D-glucosaminidase (NAG), beta2 microglobulin (beta-2 MG), creatinine (Cr), uric acid (UA), and calcium, markers of glomerular filtration blood urea nitrogen (BUN), serum Cr, UA, and parameters of oxidative damage in erythrocyte were studied in 79 Pb-exposed adolescent and 71 healthy control subjects. Blood lead levels and ALAD index were found significantly higher in the study group than that of normal control group. BUN, UA level, and glomerular filtration rates were detected in normal range in the lead-exposed group. Urinary NAG excretion and calciuria were higher in the study group than in controls. Urinary excretion of NAG was positively correlated with the blood lead levels (r=0.427). There was no relationship between blood lead levels and UA or beta-2 MG in urine. Malondialdehyde and glutathione peroxidase levels were significantly elevated in lead-exposed adolescents than controls, but changes in the catalase and superoxide dismutase activities in lead-exposed adolescents did not reach statistical significance. In conclusion, chronic low dose lead exposure seems as a cause of subtle renal impacts in adolescent workers of auto repair workshops. Lead-induced oxidative stress in erythrocytes probably contributes to these subclinical renal effects. PMID- 15526094 TI - Methadone and nicotine self-administration in humans: a behavioral economic analysis. AB - RATIONALE: Prior research has revealed inconsistencies in the behavioral relations between nicotine and opiates among methadone-maintained patients. OBJECTIVES: The current study examined whether the drug reinforcers cigarette puffs and methadone were economic complements or substitutes. METHODS: Five methadone-maintained, nicotine-dependent participants were trained to self administer methadone, cigarette puffs, or concurrently available methadone and puffs. Following training, the fixed ratio (FR) value ("price") was increased across sessions (FR 32, 64, 128, 256, and 512), first for methadone and then for puffs. Subsequently, methadone and puffs were concurrently available, and the price of each drug was increased independently, while the price of the alternative (puffs or methadone) remained constant at FR 32. RESULTS: Demand for methadone and cigarette puffs decreased as a function of increases in methadone and cigarette puff prices, respectively. When methadone and puffs were concurrently available, an increase in methadone's price decreased puff consumption, and demand for methadone was less elastic than when puffs were not concurrently available. An increase in puff price decreased puff and methadone demand, but the elasticity of puff demand was unaffected. The concurrent presence of methadone had no effect on the elasticity of demand for cigarette puffs. CONCLUSIONS: Methadone and cigarette puffs appear to be asymmetric economic complements. PMID- 15526095 TI - Bilateral Wallerian degeneration of the middle cerebellar peduncles due to paramedian pontine infarction: MRI findings. AB - Wallerian degeneration is a frequent finding in lesions of the pyramidal tract, but has been observed after damage of the other fibre systems as well. Few reports exist about Wallerian degeneration of cerebellar fibres after distant lesions to the axons. Here, we report on a patient who developed degeneration of both middle cerebellar peduncles after a paramedian pontine infarction. PMID- 15526096 TI - Genomic analysis reveals a duplication of eight rather than seven short consensus repeats in primate CR1 and CR1L: evidence for an additional set shared between CR1 and CR2. AB - We report the discovery of previously unrecognised short consensus repeats (SCRs) within human and chimpanzee CR1 and CR1L. Analysis of available genomic, protein and expression databases suggests that these are actually genomic remnants of SCRs previously reported in other complement control proteins (CCPs). Comparison with the nucleotide motifs of the 11 defined subfamilies of SCRs justifies the designation g-like because of the close similarity to the g subfamily found in CR2 and MCP. To date, we have identified five such SCRs in human and chimpanzee CR1, one in human and chimpanzee CR1L, but none in either rat or mouse Crry in keeping with the number of internal duplications of the long homologous repeat (LHR) found in CR1 and CR1L. In fact, at the genomic level, the ancestral LHR must have contained eight SCRs rather than seven as previously thought. Since g like SCRs are found immediately downstream of d SCRs, we suggest that there must have been a functional dg set which has been retained by CR2 and MCP but which is degenerate in CR1 or CR1L. Interestingly, dg is also present in the CR2 component of mouse CR1. The degeneration of the g SCR must have occurred prior to the formation of primate CR1L and prior to the duplication events which resulted in primate CR1. In this context, the apparent conservation of g-like SCRs may be surprising and may suggest the existence of mechanisms unrelated to protein coding. These results provide examples of the many processes which have contributed to the evolution of the extensive repertoire of CCPs. PMID- 15526098 TI - Vancomycin-resistant Staphylococcus aureus in a bone marrow transplantation unit. PMID- 15526099 TI - Cerebral venous thrombosis in a patient with Evans syndrome: a rare association. AB - In this report we describe a case of extensive cerebral venous thrombosis in a patient with Evans syndrome. A 19-year-old male patient with Evans syndrome was admitted to the hospital with the complaints of headache, convulsive seizure, and vomiting. The cerebral venous thrombosis including left lateral, left sigmoid, straight sinus, and vena jugularis interna was diagnosed by cerebral magnetic resonance angiography. When the thrombosis developed, he was in hematological remission and he was not receiving any medications except lamivudine for chronic hepatitis B infection. As a genetic prothrombotic risk factor, he had heterozygous prothrombin G20210A gene mutation. His clinical and radiologic findings improved after unfractionated heparin and subsequently with coumadin therapy. On follow-up, cerebral venous thrombosis reoccurred in different localizations, but complete recanalization could be obtained with antithrombotic therapy. We present the case since the association of cerebral venous thrombosis and Evans syndrome is very rare. PMID- 15526100 TI - High-resolution ultrasonography in gastrointestinal graft-versus-host disease. AB - Gastrointestinal graft-versus-host disease (GVHD) is one of the main causes of therapy-related death after allogeneic hemopoietic stem cell transplantation. Early diagnosis and immediate treatment are probably essential for improving clinical outcome. High-resolution sonography allows the evaluation of single bowel wall layers throughout the small and large bowel. We report the sonographic findings of seven consecutive patients with histologically confirmed GVHD grade II-IV of the gastrointestinal tract. The patients were examined sonographically within the 1st week after onset of clinical symptoms. Dilatation of the colon and striking bowel wall thickening were early and common features in these patients. Sonography may become a useful tool for diagnosis of patients with gastrointestinal GVHD. Its role for preclinical diagnosis, grading of severity of disease, and guiding therapy of patients with gastrointestinal GVHD warrants evaluation in prospective studies. PMID- 15526101 TI - Radiological assessment of penile prosthesis: the role of magnetic resonance imaging. AB - Penile implants offer a dependable way of restoring erections in virtually all motivated patients. The satisfaction rate among both patients and partners using these devices is high. Advances in technology have reduced the infection rate and increased the mechanical reliability of these products. However, too often, urologists do not present this option with the same authority as other treatments. The reason is fear of complications and lack of expertise in managing them. Although they are not very frequent, complications may be catastrophic. The most significant postoperative complication associated with the implant surgery is infection of the device, which is quite frequent, but some other important complications are distal and proximal perforation of the albuginea, SST deformity, "S-shaped" deformity of the penis, erosion of a component, and mechanical malfunction of the device. The best way to manage complications is to prevent them, but we do not have many diagnostic tools available. Diagnosis is based on clinical history and physical examination, but imaging techniques are also needed to explore the prosthesis "in situ" to plan the surgical approach if it is needed. In this article we review the different imaging techniques used for the diagnosis of complications of prosthetic surgery of the penis, including conventional radiology, use of sonography, the role of CT scan and the magnetic resonance imaging (MRI) of the penile prosthesis. We conclude that MRI is the most valuable method for the diagnosis of penile prosthesis complications. It is not an ionizing radiation imaging method and has the unique feature among imaging techniques of demonstrating penile anatomy in three orthogonal planes. It is superior to any other imaging method in the definition of soft tissue contrast. PMID- 15526102 TI - [Anticoagulative therapy in patients with exudative age-related macular degeneration: acute angle closure glaucoma after massive intraocular hemorrhage]. AB - BACKGROUND: Acute secondary angle closure glaucoma after massive vitreous and subretinal hemorrhage from exudative age related macular degeneration is a seldom, but devastating complication of an anticoagulative therapy. PATIENTS: Four patients (age range 70-76 years) developed unilateral acute angle closure glaucoma after massive intraocular bleeding due to exudative age-related macular degeneration and anticoagulative therapy. Three patients received Phenprocomoun and one patient thrombolytic therapy with heparin. One eye had led to blindness at initial investigation. Pars plana vitrectomy was performed in the remaining three eyes. Within a few months, 2 of these 3 eyes were blind, one eye had light perception and all three eyes developed hypotonia. CONCLUSION: Acute angle closure glaucoma after intraocular bleeding may be the initial clinical manifestation of overtreatment with anticoagulants. Early pars plana vitrectomy probably will increase the surveillance rate of the eye but not the visual prognosis. PMID- 15526104 TI - 2-Benzyloxybenzaldehyde inhibits formyl peptide-stimulated increase in intracellular Ca2+ in neutrophils mainly by blocking Ca2+ entry. AB - 2-Benzyloxybenzaldehyde (CCY1a) inhibited the formyl-Met-Leu-Phe (fMLP)-induced elevation of cytosolic [Ca(2+)]() ([Ca(2+)](i)) in rat neutrophils. The late plateau phase, but not the initial Ca(2+) spike, of the fMLP-induced [Ca(2+)](i) change was inhibited by CCY1a. In the absence of external Ca(2+), CCY1a had no appreciable effect on either the fMLP- or cyclopiazonic acid (CPA)-induced [Ca(2+)](i) elevation. CCY1a failed to inhibit [Ca(2+)](i) changes induced by N ethylmaleimide, GEA3162, ionomycin or sphingosine, but slightly inhibited the Ca(2+) signals elicited by ATP or interleukin-8 (IL-8). In a classical Ca(2+) readdition protocol, addition of CCY1a after cell activation strongly inhibited the [Ca(2+)](i) response to fMLP, whilst that to CPA was only slightly reduced. CCY1a nearly abrogated the fMLP-stimulated Mn(2+) influx but was less effective on the CPA-induced response. CCY1a attenuated the levels of tyrosine phosphorylated bands in the 70-85 kDa molecular mass range. CCY1a had no effect on the basal [Ca(2+)](i) level, the pharmacologically isolated plasma membrane Ca(2+)-ATPase activity or on the mitochondrial membrane potential. Thus, CCY1a blocks fMLP-induced Ca(2+) entry into neutrophils probably by blocking the relevant Ca(2+) channel directly or, alternatively, indirectly through the attenuation of tyrosine phosphorylation of some cellular proteins. PMID- 15526105 TI - Urotensin II stimulates plasma extravasation in mice via UT receptor activation. AB - The peptide urotensin II (U-II) is the cognate ligand of the G-protein coupled receptor UT (formerly GPR14). A role in the regulation of cardiovascular functions has been proposed for this novel peptide/receptor system. In the present study, we evaluated the ability of U-II to induce plasma extravasation in mice and attempted to characterize the receptor involved using the novel UT receptor ligand, [Orn(8)]U-II. The Evans blue technique was used to quantify plasma extravasation. U-II (0.01, 0.1, 1 and 10 nmol/kg) dose-dependently stimulated plasma extravasation in airways, gastrointestinal and urogenital tract tissues from mice, but not in the skin. In most tissues, the dose/response curves to U-II were bell shaped with the maximal effect induced by 1 nmol/kg. [Orn(8)]U II at 30 nmol/kg was per se either inactive or produced a non-significant increase in plasma extravasation; in the presence of 30 nmol/kg [Orn(8)]U-II, the effects of 1 nmol/kg U-II were always reduced and, in some tissues, abolished. The present findings demonstrate that U-II promotes plasma extravasation in various mouse vascular regions via activation of UT receptors. The mouse plasma extravasation assay will be a useful tool in future studies aimed at characterizing the pharmacological features of novel UT receptor ligands in vivo. PMID- 15526106 TI - Involvement of AP-2 binding sites in regulation of human beta-glucuronidase. AB - The lysosomal hydrolase beta-glucuronidase (beta-gluc) can be used for the bioactivation of non-toxic glucuronide prodrugs of anticancer agents. The enzyme is present at high levels in many tumours and hence may lead to an enhanced drug targeting by tumour-selective release of the active anticancer drug. Individual expression and regulation of this enzyme is one factor modulating the bioactivation of glucuronide prodrugs. Nevertheless, in contrast to murine beta gluc, which is inducible by androgens, the human enzyme has been regarded as an unregulated housekeeping gene due to a lacking TATA box and high G+C contents within the putative promotor sequence. Despite these facts, we were able to demonstrate downregulation of human beta-gluc expression by the calcium ionophore A23187 and the calcium ATPase inhibitor thapsigargin in the human hepatoma cell line HepG2. However, cis-acting elements responsible for this regulation have not yet been identified. We therefore characterised the 5'-untranslated region of the human beta-gluc gene using transient transfection assays with promotor-luciferase constructs in HepG2 cells and cloned fragments between 3,770 bp and 107 bp. A23187 reduced the beta-gluc promotor activity. This effect disappeared using fragments smaller than 356 bp. Using site-directed in vitro mutagenesis and gel electrophoretic-mobility shift assays, we found evidence of an involvement of transcription factor activating protein-2 (AP-2) binding sites on the regulation of human beta-glucuronidase by A23187. Our studies provide a basis for the understanding of the transcriptional regulation of the human beta-glucuronidase gene and could be useful for the optimisation of glucuronide prodrug therapy. PMID- 15526107 TI - Beta 2-adrenoceptor-mediated intrinsic sympathomimetic activity of carteolol: an in vivo study. AB - The intrinsic sympathomimetic activity (ISA) of a beta-adrenoceptor blocker can be mediated by beta(1)- or beta(2)-adrenoceptors. The aim of this study was to characterize the ISA of the beta-adrenoceptor blocker carteolol in healthy volunteers. Two approaches were employed. First, we assessed the effects of carteolol (20, 40 or 80 mg p.o.) on blood pressure, heart rate and heart-rate corrected duration of electromechanical systole (QS(2)c, a measure of cardiac contractility) in the volunteers. Carteolol dose-dependently increased systolic blood pressure, heart rate and contractility and decreased diastolic blood pressure. The beta(1)-adrenoceptor blocker bisoprolol did not attenuate these carteolol effects, but rather enhanced the effects on heart rate and systolic blood pressure. Second, we treated volunteers for 7 days with 1 x 20 mg/day carteolol and assessed lymphocyte beta(2)-adrenoceptor density (by (-)-[(125)I] iodocyanopindolol binding) and functional responsiveness (by 10 muM isoprenaline induced increase in lymphocyte cyclic AMP content). Carteolol significantly reduced lymphocyte beta(2)-adrenoceptor density and function. After withdrawal of carteolol lymphocyte beta(2)-adrenoceptor density and function recovered only very slowly and had not returned to control levels 11 days after carteolol withdrawal. In conclusion, the fact that, on the one hand, the cardiovascular effects of carteolol were not attenuated by the beta(1)-adrenoceptor blocker bisoprolol and, on the other, carteolol significantly decreased lymphocyte beta(2)-adrenoceptor density and function is in favour of the idea that the ISA of carteolol is mediated by beta(2)-adrenoceptors. Involvement of an additional receptor site (e.g. the propranolol-resistant state of the beta(1)-adrenoceptor), however, cannot be excluded. PMID- 15526108 TI - Differential amino acid transmission in the locus coeruleus of Wistar Kyoto and spontaneously hypertensive rats. AB - In addition to differences in their blood pressure, Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR) are known to differ in their emotional behaviour. The neurochemistry underlying these differences is not well understood. In the present study the release rates of the two main regulatory amino acids in the locus coeruleus, glutamate and gamma-aminobutyric acid (GABA), were monitored in WKY rats and SHR to investigate whether basal and/or challenged neurotransmission differs between these strains. The strains differed in their basal blood pressure (WKY 102+/-2 mmHg, SHR 140+/-4 mmHg), as well as in their emotional behaviour, since WKY rats displayed enhanced anxiety-related behaviour in the open field test (time in centre: WKY 197+/-40 s/30 min, SHR 741+/-93 s/30 min). Basal glutamate and GABA release rates did not differ between WKY rats and SHR. A rise in blood pressure induced by intravenous infusion of noradrenaline for 10 min enhanced GABA release in WKY rats by 60%, while no effect was observed in SHR. Glutamate release did not respond to experimental hypertension in both strains. Intravenous infusion of sodium nitroprusside led to a fall in blood pressure, which was less pronounced and was of shorter duration in WKY rats than in SHR. The depressor response had no effect on amino acid release in the locus coeruleus of both strains. Mild stress induced by noise or tail pinch led to slight rises in arterial blood pressure (10 mmHg and 20 mmHg respectively), which were similar in WKY rats and SHR. Tail pinch enhanced the release rates of glutamate and GABA in the locus coeruleus of WKY rats and SHR; however, no strain differences were noted. Noise stress did not significantly influence amino acid release. These findings demonstrate that SHR and WKY rats differ in GABAergic neurotransmission, which is revealed in response to specific cardiovascular challenges, but not to mild stressors. The observed lack of GABA response to blood pressure elevation in SHR may reflect a disturbed mechanism counteracting high blood pressure, possibly contributing to hypertension in this strain. PMID- 15526109 TI - A comparison between haemodynamic effects of vasopressin analogues. AB - Some analogues of arginine vasopressin (AVP) reportedly possess hypotensive properties, and two such peptides are Cys(1)-Tyr(2)-Phe(3)-Val(4)-Asn(5)-Cys(6) Pro(7)- d-Arg(8)-Gly(9)-NH(2) (VD-AVP) and d(CH(2))(5)-Cys(1)- d-Tyr(Et)(2) Arg(3)-Val(4)-Asn(5)-Cys(6)-Lys(7)-Lys(8)-ethylenediamine(9) (TA-LVP). In the present investigation we examined the effects of TA-LVP (0.3, 1.0 and 3.0 microg/kg/min), VD-AVP (0.3, 1.0 and 3.0 microg/kg/min) and AVP (1.0, 3.0, 10 ng/kg/min) on haemodynamics, blood volume (BV) and plasma troponin levels in anaesthetised rats. Infusion of TA-LVP significantly ( P<0.05) reduced blood pressure (-45+/-3%; n=8; mean +/- SEM), mean circulatory filling pressure ( P(mcf); -41+/-3%), and cardiac output (CO; -59+/-4%). The reduction in CO at a lower dose of TA-LVP was due to reduced venous tone, while at higher doses the reduction was predominantly the result of reduced BV (-35+/-4%). The large decrease in BV during the infusion of TA-LVP, substantially increased resistance to venous return (50+/-11%), which was the main contributor in reducing CO. Administration of AVP significantly increased blood pressure (41+/-4%) and arterial resistance (98+/-16%) without any impact on P(mcf) and BV, while significantly reducing CO (-26+/-5%). Infusion of VD-AVP did not produce hypotension, but produced a modest but significant reduction in CO (-18+/-5%) and insignificant but moderate increases in peripheral resistance (30+/-12%) and resistance to venous return (28+/-8%). Plasma troponin levels were not affected by any of the peptides. The hypotensive action of TA-LVP was due to a reduction in CO as a result of a reduced pre-load, while the pressor effect of AVP increased after-load sufficiently to impede flow, reducing CO. VD-AVP was devoid of any hypotensive effects, suggesting that V(2)-vasopressin receptors are most likely to play a limited role in the control of cardiac and vascular function in these animals. PMID- 15526110 TI - Three-dimensional isobolographic analysis of interactions between lamotrigine and clonazepam in maximal electroshock-induced seizures in mice. AB - The anticonvulsant effects of lamotrigine (LTG) and clonazepam (CZP) and combinations thereof against maximal electroshock (MES)-induced seizures in mice were investigated using three-dimensional (3D) isobolographic analysis. With this method, the doses of fixed-ratio combinations of the drugs (1:3, 1:1 and 3:1) that elicited 16, 50 and 84% of the maximum anticonvulsant effect were determined. Additionally, to evaluate the characteristics of interactions observed with 3D isobolography, the brain concentrations of both drugs were verified pharmacokinetically. The 3D isobolographic analysis showed that LTG and CZP combined at the fixed ratios of 3:1 and 1:1 interacted synergistically in the MES test for all anticonvulsant effects between 16% and 84% of maximum. In contrast, the combination of LTG and CZP at the fixed ratio of 1:3 showed only pure additivity for all estimated effects in 3D isobolography. Moreover, none of the examined antiepileptic drugs altered the brain concentrations of the coadministered drug, so the observed interactions in the MES test are of a pharmacodynamic nature. The 3D isobolographic findings suggest that in epilepsy therapy, increased efficacy of seizure control (synergistic interaction) might be achieved by using LTG and CZP in combination. In this study, some important problems and assumptions related to statistical analysis of data in 3D isobolography are discussed. PMID- 15526111 TI - Reversal of loop ileostomy under spinal anaesthesia. AB - BACKGROUND AND AIMS: Traditionally, loop ileostomy is reversed under general anaesthetic. In patients with severe cardiorespiratory disease, many surgeons opt for a one-stage, low Hartmann's procedure to avoid a second general anaesthetic to reverse a defunctioning stoma. Closure of loop ileostomy under spinal anaesthetic would allow high-risk patients to avoid a permanent stoma and a general anaesthetic. PATIENTS AND METHODS: Seven patients (6 of whom were men) had reversal of loop ileostomy under spinal anaesthetic. The median age was 63 years (range 43--70). Six patients had significant co-morbidity with a median American Society of Anesthesiologists (ASA) grade of 3. The ileostomy was reversed in a side-to-side fashion using a linear stapler. The small bowel wall was infiltrated with local anaesthetic prior to firing the stapler. RESULTS AND FINDINGS: All patients tolerated the procedure without discomfort. Patients started feeding on the first postoperative day. Analgesia requirements postoperatively were minimal. No complications occurred due to the anaesthetic technique. CONCLUSION: With careful patient selection, preparation and a gentle and meticulous surgical technique, reversal of loop ileostomy can be achieved under spinal anaesthesia, thereby, saving high-risk patients with low tumours (suitable for sphincter preservation) from having a one-stage resection with permanent stoma. PMID- 15526112 TI - Comparison study between electrogalvanic stimulation and local injection therapy in levator ani syndrome. AB - BACKGROUND AND AIMS: Two theories have been reported for the pathophysiology of levator ani syndrome: the spastic cycle hypothesis and the local inflammation (Tendinitis) hypothesis. This study compared two treatment modalities in order to determine which of the two hypotheses is more appropriate. SUBJECTS AND METHODS: In this prospective study, Group EGS (n=22) underwent electrogalvanic stimulation twice a week. Group LI (n=31) underwent a local injection of a 40-mg triamcinolone acetonide mix with 1 ml 2% lidocaine into the maximal tender point of the arcus tendon in the levator ani muscle. RESULTS: The most common location of tenderness was the left anterior of the arcus tendon of the levator ani muscle. At the last follow-up (12 months), the LI group showed more relief, more improvement, and fewer failures than the EGS group. No difference was seen between the mean pain scores (verbal analog scale: 0-100) of the two groups at either the 1-week or the 12-month follow-up. However, the LI group showed better results at the 1-month, 3-month, and 6-month follow-ups. CONCLUSION: The LI group showed better short-term results than the EGS group. Therefore, the tendinitis hypothesis seems to be the more reliable one for levator ani syndrome. However, because the subjective responses of the patients indicated that a sufficient level of patient satisfaction had not been achieved, we cannot positively conclude that the tendinitis hypothesis is the more reliable one for the pathophysiology of levator ani syndrome. PMID- 15526114 TI - Regional differences of myocardial infarct development and ischemic preconditioning. AB - The spatial and temporal development of myocardial infarction depends on the area at risk (AAR), the severity and duration of blood flow reduction (energy supply) as well as on heart rate and regional wall function (energy demand). Both supply and demand can vary within the AAR of a given heart, potentially resulting in differences in infarct development. We therefore retrospectively analyzed infarct size (IS, %AAR, TTC) in 24 anesthetized pigs in vivo following 90 min hypoperfusion and 120 min reperfusion of the LAD coronary artery, which supplies parts of the LV septum (LVS) and anterior free wall (LVAFW). The total LAD perfusion territory averaged 49.8 +/- 14.2 (SD) g (49.2 +/- 8.4% of LV); 61.4 +/- 8.1% of the AAR was LVAFW. IS within the LVS was 25.3 +/- 15.1%, while IS within the LVAFW was 16.6 +/-10.1% (p<0.05). While ischemic blood flow (radiolabeled microspheres) did not differ between LVS (0.05 +/- 0.02 ml/min/g) and LVAFW (0.05 +/- 0.03 ml/min/g), perivascular connective tissue (56 +/- 9 vs. 38+/-7 microm(2), p < 0.05) and the capillary-to-myocyte distance (1.65 +/- 0.23 vs. 1.18 +/- 0.23 mm, p < 0.05) were larger in LVS than in LVAFW. Interestingly, IS in LVS (9.3 +/- 9.6%, n = 24) and LVAFW (9.2 +/- 9.1%) were reduced to the same absolute extent by ischemic preconditioning with one cycle of 10 min ischemia and 15 min reperfusion, suggesting that a similar regional difference exists also in the protection afforded by ischemic preconditioning. The mechanism(s) for that remain(s) to be established. CONCLUSION: In pigs, regional differences in infarct development and protection from it exist in the LAD perfusion territory, which are independent of ischemic blood flow but apparently related to pre-existing structural differences. PMID- 15526113 TI - Long-term results of botulinum toxin for the treatment of chronic anal fissure: prospective clinical and manometric study. AB - BACKGROUND: The aim of this prospective trial was to analyse the effectiveness and morbidity of chemical sphincterotomy in the treatment of chronic anal fissure after a 3-year follow-up. METHODS: One hundred consecutive patients with chronic anal fissures were treated by chemical sphincterotomy with 25 U botulinum toxin injected into the internal sphincter. Clinical and manometric results were recorded. RESULTS: No major complications were found; initial incontinence at the 2-month review (6%) spontaneously reversed at 6 months. There was a tendency of progressive recurrence over time, with an overall healing after 3 years of 47%. We found a group of patients with clinical (symptoms longer than 12 months and presence of a sentinel pile before treatment) and manometric factors (persistently elevated mean resting pressure, percentage of time with slow waves, and number of patients or percentage of time with ultra slow waves after treatment) associated with a higher recurrence of anal fissures. CONCLUSION: Since it avoids the greater risk of incontinence associated with surgical sphincterotomy, we recommend the use of botulinum toxin as the first therapeutic approach for patients with chronic anal fissure and risk factors for incontinence; despite the higher rate of recurrence associated with this treatment. In patients with factors related to recurrence, re-injection with higher doses of botulinum toxin or complementary medical-surgical treatment should be considered. PMID- 15526115 TI - Electrical remodeling of the canine superior vena cava after chronic rapid atrial pacing. AB - BACKGROUND: The superior vena cava (SVC) might serve as the trigger and/or substrate for paroxysmal atrial fibrillation (AF). However, the electrophysiological properties of the SVC with chronic AF are unknown. The purposes of this study were to investigate the electrophysiological properties of the SVC and the electropharmacological effects of intravenous dl-sotalol on the canine SVC after chronic rapid atrial pacing (RAP). METHODS AND RESULTS: In the control group, the effective refractory period (ERP), conduction velocity, and AF inducibility of the SVC were assessed in 6 normal dogs before and after an infusion of dl-sotalol. In the experimental group, the ERP, conduction velocity, and AF inducibility of the SVC were assessed before and after dl-sotalol administration in 10 dogs after 8 weeks of RAP. The SVC showed a shorter ERP, decreased slope of rate-adaptation of the ERP, increased ERP dispersion, a decreased conduction velocity, and increased inducibility and duration of AF initiated from the SVC in the RAP dogs. In the RAP dogs, intravenous dl-sotalol significantly increased the ERP, but dl-sotalol did not change the slope of rate adaptation of the ERP, dispersion of the ERP, conduction velocity, inducibility, or duration of AF initiated from the SVC. CONCLUSIONS: The present study demonstrates that the canine SVC shows significant electrical remodeling and increased AF vulnerability after chronic RAP. Intravenous dl-sotalol was unable to decrease the inducibility or duration of AF initiated from the SVC. PMID- 15526116 TI - The temporal relationship between p38 MAPK and HSP27 activation in ischaemic and pharmacological preconditioning. AB - An ischaemic preconditioning protocol and subsequent sustained ischaemia were characterized by activation and attenuation of p38 MAPK phosphorylation, respectively. However, the significance of events downstream of p38 MAPK needs investigation. Therefore the temporal relationship between phosphorylation of p38 MAPK and its downstream substrate HSP27 was studied during either an ischaemic or beta-adrenergic preconditioning protocol and during sustained ischaemia. Isolated rat hearts were preconditioned (with or without a p38 MAPK inhibitor, SB203580) with 1 x 5 min or 3 x 5 min global ischaemia or 5 min beta-adrenergic stimulation (10(-7) M isoproterenol), followed by 25 min sustained ischaemia and 30 min reperfusion. Hearts were freeze-clamped at different time intervals and fractionated to determine p38 MAPK and HSP27 phosphorylation, via Western blotting. Significant phosphorylation of cytosolic p38 MAPK and membrane (myo fibrillar) HSP27 occurred at the end of the first preconditioning episode. However, p38 MAPK phosphorylation disappeared during subsequent preconditioning episodes, while HSP27 phosphorylation was maintained for the duration of the protocol. Similar changes in p38 MAPK and HSP27 occurred with 5 min beta adrenergic preconditioning. After 25 min ischaemia, significant phosphorylation of cytosolic and membrane HSP27 was observed, while p38 MAPK phosphorylation was attenuated in ischaemic and beta-adrenergic preconditioned compared to non preconditioned hearts. SB203580-induced abolishment of p38 MAPK and HSP27 phosphorylation during the triggering phase of both preconditioning protocols reversed the changes in these parameters seen after sustained ischaemia. The results suggest that p38 MAPK activation triggers HSP27 phosphorylation during both the preconditioning protocols and during sustained ischaemia. Protection of preconditioned hearts during sustained ischaemia was characterized by phosphorylation of both cytosolic and myofibrillar HSP27. PMID- 15526117 TI - Cross sections of electron inelastic interactions in DNA. AB - The cross sections of electron inelastic interaction in DNA are calculated using the dielectric response theory and Penn statistical approximation, with the exchange correction included. An empirical approach to obtain optical energy loss function is given for the organic compounds without available optical data. Comparisons of the calculated data with available experimental and theoretical results have been done to show the reliability of the approach proposed in this work. Using this approach, the total inelastic cross sections for five bases: guanine, adenine, thymine, cytosine and uracil have been calculated in the energy range of E< or =10 keV and compared with those recently obtained with the Deutsch Mark formalism and the Binary-Encounter-Bethe model, respectively. An equivalent unit of the DNA molecule is constructed according to the contents of A-T and G-C base pairs in DNA, and is divided into five constituents, i.e. sugar-phosphate and four bases. The total inelastic cross sections for the constructed unit of the DNA molecule and its constituents have also been calculated. PMID- 15526119 TI - Within-trophic group interactions of bacterivorous nematode species and their effects on the bacterial community and nitrogen mineralization. AB - Knowledge of the interactions between organisms within trophic groups is important for an understanding of the role of biodiversity in ecosystem functioning. We hypothesised that interactions between bacterivorous nematodes of different life history strategies would affect nematode population development, bacterial community composition and activity, resulting in increased N mineralization. A microcosm experiment was conducted using three nematode species (Bursilla monhystera, Acrobeloides nanus and Plectus parvus). All the nematode species interacted with each other, but the nature and effects of these interactions depended on the specific species combination. The interaction between B. monhystera and A. nanus was asymmetrically competitive (0,-), whereas that between B. monhystera and P. parvus, and also A. nanus and P. parvus was contramensal (+, -). The interaction that affected microcosm properties the most was the interaction between B. monhystera and P. parvus. This interaction affected the bacterial community composition, increased the bacterial biomass and increased soil N mineralization. B. monhystera and P. parvus have the most different life history strategies, whereas A. nanus has a life history strategy intermediate to those of B. monhystera and P. parvus. We suggest that the difference in life history strategies between species of the same trophic group is of importance for their communal effect on soil ecosystem processes. Our results support the idiosyncrasy hypothesis on the role of biodiversity in ecosystem functioning. PMID- 15526120 TI - Resectable hilar cholangiocarcinoma: surgical treatment and long-term outcome. AB - Unresected hilar cholangiocarcinoma has a dismal prognosis, but advances in staging and surgical techniques have given well-selected patients a chance of long-term survival if curative resection is possible. This review summarizes the state of the art in diagnosis, treatment, and outcome for patients with biliary obstruction at the hilus of the liver. PMID- 15526121 TI - Prognosis after reoperation for local recurrence of papillary thyroid carcinoma. AB - PURPOSE: To investigate the factors associated with a favorable prognosis after reoperation for local recurrent papillary thyroid carcinoma (PTC), we reviewed 45 patients who underwent surgery for first local recurrence of PTC. METHODS: We divided the patients into two groups. Group A (n = 28) had no second recurrence, and group B (n = 17) had second local recurrence after surgery for recurrence. RESULTS: The mean follow-up period after reoperation was 56.9 months. The mean age at the time of reoperation in group A was significantly lower than that in group B, at 48.1 years versus 62.3 years, respectively (P = 0.0007). The mean age at the time of the initial operation in group A was also significantly lower than that in group B, at 40.1 years versus 55.1 years, respectively (P = 0.0006). Patients with recurrent tumors only outside the area dissected at the initial operation (n = 27) had a better outcome than those with recurrence within the dissected area (n = 18; P = 0.0127). Patients who underwent systematic partial or modified neck dissection (n = 36) had a better outcome than those who underwent only simple local resection (n = 9; P = 0.0169). CONCLUSION: For local recurrent PTC, systematic neck dissection is recommended over local resection of recurrent tumors. PMID- 15526122 TI - Serum tissue inhibitors of metalloproteinase-1 and -2 in patients with non-small cell lung cancer. AB - PURPOSE: In view of the increasing number of patients diagnosed with lung cancer every year worldwide, there is an urgent need for an effective screening marker to improve its early detection. METHODS: We quantified the level of immunoreactive proteins for the tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 in the sera of 54 patients with non-small cell lung cancer (NSCLC) and 25 healthy control subjects, by using enzyme-linked immunosorbent assays with specific monoclonal antibodies. RESULTS: The TIMP-1 level was significantly higher, and the TIMP-2 level was significantly lower in the patients with NSCLC than in the controls. Furthermore, both TIMP-1 and TIMP-2 were significantly higher in patients with squamous cell carcinoma than in those with adenocarcinoma. The TIMP-1 level in patients with stage III/IV disease was significantly higher than that in those with stage I/II disease. The TIMP-1/TIMP 2 ratio was significantly higher in the patients with NSCLC, and the receiver operating characteristic curves analysis revealed that the TIMP-1/2 ratio, but not TIMP-1 or -2 alone, was a better screening marker for NSCLC than carcinoembryonic antigen (P < 0.0001). Patients with a high TIMP-1 value had significantly shorter disease-free survival (P = 0.0479), but those with a high TIMP-1/2 ratio did not. CONCLUSION: The TIMP-1/2 ratio may be a good screening marker of NSCLC; however, it was less effective than TIMP-1 as a prognostic factor. PMID- 15526123 TI - Endoscopic hemostasis using fibrin adhesive to treat hemorrhage in the upper digestive system. AB - PURPOSE: There are several methods of achieving endoscopic hemostasis of hemorrhage in the upper digestive system. We compared the therapeutic results and advantages of using a local injection of fibrin adhesive for endoscopic hemostasis, which we have found more effective than other hemostatic methods. METHODS: Between October 2000 and April 2002, 16 patients with hemorrhage in the upper digestive system underwent endoscopic hemostasis using fibrin adhesive. The hemorrhage was caused by a hemorrhagic tendency from liver disease, anticoagulant therapy, or failed hemostasis with clipping or local ethanol injection. The fibrin adhesive was injected through a standard 21-gauge endoscopic needle using the so-called sandwich method. RESULTS: Hemostasis was successfully achieved by a single local injection of fibrin adhesive, in all except one patient who had been on anticoagulant therapy for a long time and needed an additional local injection of fibrin adhesive. CONCLUSION: Fibrin adhesive does not cause any tissue injury, and a sufficient amount can be injected endoscopically even in patients with liver dysfunction and those on anticoagulant therapy. Thus, we think that endoscopic hemostasis with fibrin adhesive is safe and effective. PMID- 15526124 TI - Potentially curative resection for locoregional recurrence of colorectal cancer. AB - PURPOSE: Local recurrence after curative surgery for colorectal cancer may be treated by potentially curative surgery, defined as resection of all macroscopic disease. We conducted this retrospective study to show the effectiveness of potentially curative resection for patients with locoregional recurrence (LRR). METHODS: We reviewed the records of 242 patients who underwent curative resection of colorectal cancer in our unit between 1988 and 2000. Locoregional recurrence developed in 50 (20.6%) patients, and was treated by R0 resection in 10 (20%) patients (group R0), by R1 resection in 8 (16%) patients (group R1), by R2 resection in 13 (26%) patients (group R2), and by surgery without resection in 12 (24%) patients (group NR). Seven (15%) patients did not undergo surgery (group NS). RESULTS: The mean survival periods were 48, 36, 10, 5.6, and 5 months in groups R0, R1, R2, NR, and NS, respectively. There was no significant difference in survival between groups R0 and R1 (P = 0.5), but survival was significantly longer in groups R0 and R1 than in groups R2, NR, and NS (P = 0.001). CONCLUSION: These findings show that surgeons should aim to achieve at least macroscopic clearance of the recurrent tumor. Potentially curative surgery improves survival in selected patients with LRR after curative resection of colorectal cancer. PMID- 15526125 TI - Preoperative serum hyaluronic acid level as a good predictor of posthepatectomy complications. AB - PURPOSE: We evaluated the efficiency of measuring hyaluronic acid (HA) levels preoperatively in patients with injured liver disease as a predictor of complications after hepatectomy. METHODS: We examined patients who underwent hepatectomy for liver tumors secondary to chronic viral liver diseases or obstructive jaundice. RESULTS: The preoperative HA level correlated significantly with the indocyanine green retention rate at 15 min, liver activity at 15 min by technetium-99m galactosyl human serum albumin scientigraphy, and the histopathological activity index. It was also significantly elevated in patients with severe fibrosis caused by cirrhosis. After hepatectomy, the HA level was increased on postoperative day (PODS) 7, but had normalized by POD 28. The preoperative HA level tended to correlate with the regeneration rate on POD 28, and was significantly higher in patients with prolonged ascites or hepatic failure postoperatively. Multivariate analysis identified a serum HA level above 200 or 150 ng/ml as the only significant predictor of postoperative hepatic failure or long-term ascites, respectively (P < 0.05). CONCLUSION: Our findings indicate that the preoperative serum HA level is a good predictor of postoperative complications in patients who undergo hepatectomy for injured liver disease. PMID- 15526127 TI - Clinical study of transient portal vein stenosis induced after pancreatic head resection. AB - PURPOSE: We prospectively analyzed the clinical significance of transient portal vein stenosis soon after a pancreaticoduodenectomy (PD) as detected on three dimensional computed tomography (3D-CT) for portography. METHODS: This study included 20 patients who underwent a generalized pancreatic head resection. A 3D CT scan was taken seven times up until the eighth postoperative week for each patient. The 3D images were reconstructed by the Voxel Transmission method. The portal vein images were quantified using the portal volume rates (PVR). The relationship between portal vein stenosis and six clinical factors was statistically analyzed. RESULTS: The 3D-CT scans showed portal vein stenosis in all 20 patients. The preoperative mean of these 20 PVR were 100%, and the postoperative values were 76.2%, 64.1%, 69.5%, 75.1%, 84.1%, and 89.8% at weeks 1 4, 6, and 8, respectively. Portal vein stenosis soon after PD reached a peak in the second week and almost disappeared by the eighth postoperative week. Three clinical factors, namely, the main pancreatic duct diameters, pancreatic reconstruction methods, and postoperative prognoses, significantly affected portal vein stenosis. CONCLUSION: The stenotic changes to the portal vein are considered to be significant prognostic indicators of pancreatic anastomotic trouble and postoperative complications. PMID- 15526126 TI - Intraoperative continuous external bile drainage during pancreaticoduodenectomy. AB - PURPOSE: The common hepatic duct is usually divided during the early stage of pancreaticoduodenectomy. However, abrupt, complete, and prolonged closure of the proximal common duct stump can cause liver damage in the course of this long operation, resulting in postoperative liver dysfunction and associated complications. Here, we investigate this phenomenon further. METHODS: We performed intraoperative continuous external bile drainage (IBD) in 43 consecutive patients (drainage group) and compared postoperative liver enzyme levels, morbidity including liver dysfunction, and outcomes with those of a control group (n = 41). RESULTS: There were no complications associated with IBD catheter insertion in this series. The drainage group had significantly lower transaminase levels within the first 7 postoperative days (PODs) than the control group. Postoperative liver dysfunction was confirmed in six patients from the control group and in one patient from the drainage group (P = 0.04). However, there were no significant differences between these two groups in terms of postoperative morbidity (other than liver dysfunction), relaparotomy, and in hospital mortality rates. CONCLUSION: Intraoperative continuous external bile drainage failed to improve the overall morbidity and mortality rates in this series. However, our findings showed that prolonged intraoperative complete closure of the common hepatic duct contributed to postoperative liver dysfunction in most patients and that IBD, which is easy and safe to perform, could reduce intra-operative liver damage and prevent postoperative liver dysfunction. PMID- 15526128 TI - The use of totally extraperitoneal endoscopic hernioplasty for the treatment of groin hernia. AB - PURPOSE: To examine the use of totally extraperitoneal endoscopic hernioplasty (TEP) and its postoperative complications in order to obtain better and more stable treatment results. METHODS: We studied 73 patients (83 cases) for whom TEP was performed at our hospital between June 1995 and February 2003. The types of hernias included unilateral hernia (63), 10 bilateral hernias, 47 indirect inguinal hernias, 29 direct inguinal hernias, and 7 other types. RESULTS: The average body mass index was 22.3 kg/m2. The average duration of operation was 87.1 +/- 31.1 min for unilateral hernias and 106.5 +/- 38.4 min for bilateral hernias. The length of hospitalization was 9.46 days on average, and the postoperative stay was approximately 6.21 days. 80.8% of all cases were administered analgesics for 1.3 days on average. Postoperative complications were observed in 12 cases: peritoneum injury (3), postoperative subcutaneous suggillation (2), ischemic orchitis (1), and other symptoms which required a switch to other operational techniques (7). CONCLUSION: The study revealed that patients with previous abdominal surgery and patients 70 years of age or older tended to show significantly more postoperative complications and thus, for these patients, the use of TEP should be determined only after careful consideration. PMID- 15526129 TI - Lung injury after intestinal ischemia-reperfusion may be avoided by the reduced absorption of locally produced cytokines. AB - PURPOSE: It has been suggested that inflammatory mediators such as cytokines released during intestinal ischemia and reperfusion increase permeability in the lungs. Cytokines exist at concentrations several hundred times higher at the site of inflammation than in the blood. When absorbed, the locally produced cytokines may affect multiple remote organs. We thus investigated whether the isolation of the intestine in a bag during ischemia and reperfusion can reduce subsequent lung injury. METHODS: Rats were divided into three groups: group 1, simple laparotomy (sham); group 2, intestinal ischemia and reperfusion (I/R); and group 3, intestinal ischemia and reperfusion with an intestinal bag (IB). Lung permeability was assessed using the Evans Blue leakage method. Cytokines (interleukin-1beta, tumor necrosis factor alpha, interleukin-8) in the plasma and ascites were measured by enzyme-linked immunosorbent assay. RESULTS: The increase in lung permeability of I/R significantly decreased in IB (1.73 +/- 0.48 vs 1.05 +/- 0.22, P < 0.01). The plasma cytokine concentrations were also lower in IB than in I/R. In addition, the cytokine levels in the intestinal bag fluid were extremely high. CONCLUSION: The isolation of the intestine during ischemia and reperfusion was found to reduce the degree of subsequent lung injury, possibly due to the reduced absorption of locally produced cytokines via the parietal peritoneum. PMID- 15526130 TI - The relationship of a Helicobacter heilmannii infection to the mucosal changes in abattoir and laboratory pig stomach. AB - PURPOSE: A pig ulcer model in which ulceration is reproducibly induced in the pars oesophagea (a tongue of the oesophageal squamous epithelium that extends into the pig stomach) by bile duct ligation (BDL) was used in this study to determine whether Helicobacter heilmannii (Hh) is a predisposing factor in the ulceration of this region. The infection with Hh and its relationship to ulceration and mucus integrity was examined. METHODS: We microscopically investigated the occurrence of spontaneous pars oesophageal ulceration in 33 pigs from a local abattoir and 5 pigs nurtured in pens in our surgical laboratory (JSM). Further groups of 5 and 6 JSM pigs underwent a sham operation and a BDL, respectively. Giemsa staining was used to detect Hh and purified mucin was characterized by gel filtration. RESULTS: Ten of 33 and 2 of 5 of the stomachs of abattoir and JSM pigs, respectively, were positive for Hh by Giemsa stain. Three of the 33 abattoir pigs showed ulceration in the pars oesophagea and none of these was Hh-positive. All six of the bile duct-ligated pigs showed ulceration in the pars but only 2 of these were Giemsa-positive. Only 8 of 33 of the abattoir pigs had > or =50% large polymeric mucin that was eluted in the void/excluded volume of a Sepharose 2B column. CONCLUSIONS: There was no consistent correlation between an infection of the pig stomachs by Hh, an ulceration of the pars oesophagea, and mucin degradation. There was a significant difference between the percentage of polymeric mucin from the abattoir pigs and that of the JSM group (P < 0.003), the JSM group vs sham-operated pigs (P < 0.011), and JSM vs BDL pigs (P < 0.0005), but there appeared to be no association between the infectivity with Hh and mucin degradation. PMID- 15526131 TI - Mediastinal neurofibroma originating from the left intrathoracic phrenic nerve: report of a case. AB - We report a case of mediastinal neurofibroma originating from the left phrenic nerve in a 42-year-old woman who was referred to us after a routine chest X-ray showed a smooth, round abnormal shadow in the left middle lung field adjacent to the heart. We resected a 25 x 20 x 20-mm tumor by video-assisted thoracic surgery. Histopathological examination confirmed that the lesion was a mediastinal neurofibroma originating from the left phrenic nerve without von Recklinghausen's disease. Neurogenic mediastinal tumors originating from the phrenic nerve are very rare, and to the best of our knowledge, no other case of a mediastinal neurofibroma originating from the phrenic nerve in a patient without von Recklinghausen's disease has ever been reported. PMID- 15526132 TI - Video-assisted thoracic surgery lobectomy for extralobar pulmonary sequestration in a child: report of a case. AB - Pulmonary sequestration in infants and children is conventionally treated by resecting the sequestered lung parenchyma (sequestrectomy) or by performing lobectomy through a standard thoracotomy. We performed lobectomy by video assisted thoracic surgery, using an original tracheal tube that we designed, in a 6-year-old boy with extralobar pulmonary sequestration and bronchiectasis in the left lower lobe. PMID- 15526133 TI - Repair of delayed left ventricular rupture after mitral valve replacement: report of a case. AB - Rupture of the left ventricle (LV) after mitral valve replacement (MVR) is a devastating complication, associated with high mortality. A 64-year-old woman with a type I delayed LV rupture, which occurred after MVR with a 27-mm St. Jude Medical mitral prosthesis for mitral stenosis, was successfully treated by a combination of intracardiac and extracardiac surgical repair techniques. The extracardiac repair involved approximating the edges of myocardium around the tear with large sutures bolstered by strips of Teflon felt, then covering the epicardial hematoma with another porcine pericardial patch, using gelatin resorcinol formaldehyde glue and collagen sheets. The intracardiac repair involved suturing the edges of an oval piece of porcine pericardium to the endocardium around the laceration. No LV pseudoaneurysm was detected postoperatively on echocardiography or computed tomography scans. The patient is well 2 years after the operation. PMID- 15526134 TI - Simultaneous open and endoluminal repair of ruptured abdominal and thoracic aortic aneurysms: report of a case. AB - A 66-year-old woman was transferred to our hospital for emergency treatment of a ruptured abdominal aortic aneurysm (AAA) and impending rupture of a descending thoracic aortic aneurysm (TAA) caused by a Stanford type-B dissection. She had severe coronary artery disease and a highly calcified aorta, and had been taking long-term steroids for rheumatoid arthritis. Endovascular repair of the TAA failed because the femoral artery was too small, so we performed simultaneous repair of the TAA and the AAA. A temporary axillofemoral bypass was constructed and the AAA was replaced with a bifurcated prosthetic graft. A thoracic stent graft was delivered successfully through a chimney graft of the abdominal graft. About 4 months later, the TAA extended proximally, causing hemoptysis, which was stopped by placing a new stent graft proximal to the previous one. This case report shows that a combination of open and endovascular repair is useful for treating a TAA with an AAA, especially in a small or frail patient. PMID- 15526135 TI - Using the angio-seal to achieve hemostasis in prosthetic endovascular surgery: report of three cases. AB - At least 10% of patients who undergo bilateral aortofemoral bypass are at risk of needing a reoperation for late prosthetic thrombosis because of reduced outflow as the disease progresses. To prevent occlusion of the prostheses, we performed endovascular surgery with transprosthetic access for distal stenosis. We report our experience of using the Angio-Seal with transprosthetic access after angioplasty in three patients who had undergone bilateral aortofemoral bypass. Hemostasis was achieved in all three patients. There were no complications, such as hemorrhage, hematoma, or prosthetic infection, and all three patients were discharged within 24 h. At the 12-, 15-, and 24-month follow-up, none of the patients had any sign of recurrent claudication. Using the Angio-Seal in bilateral aortofemoral bypass provided the means of treating distal stenosis by endovascular surgery with transprosthetic access. This method is both rapid and safe, and may broaden the indications for the endovascular treatment of distal arteriopathies in patients with vascular prostheses. PMID- 15526136 TI - Mucoepidermoid carcinoma of the liver diagnosed as a liver abscess: report of a case. AB - Mucoepidermoid carcinoma of the liver is a rare variant of cholangiocarcinoma, containing both mucus-secreting glandular cells and squamous cells mixed in nests. We report a case of mucoepidermoid carcinoma of the liver in a 69-year-old woman who presented with a 1-week history of fever, chills, and right flank pain. On admission, she was not jaundiced, and under a provisional diagnosis of liver abscess, a pigtail catheter was inserted into the abscess cavity. We performed right hepatectomy and partial excision of the diaphragm 1 month later. Microscopically, the tumor was composed of solid and invasive nests of epidermoid and mucin-producing cells with desmoplastic stroma. The epidermoid component of the tumor contained intercellular bridges and individual cell keratinization. Alcian blue and Periodic acid-Schiff (PAS) staining confirmed that there was mucin in the cytoplasm of mucus-secreting cells. The tumor cells, intrahepatic bile ducts, and ductules were consistently reactive with cytokeratin (CK) 7 and negative for CK20. The adjacent nonneoplastic liver cells were CK 7-/CK20-, and P63 immunostaining was positive in the epidermoid cells. The tumor was diagnosed as mucoepidermoid carcinoma arising from the intrahepatic bile duct. Despite aggressive surgical treatment, the patient died of multiple liver metastases 4 months after the right hepatectomy. PMID- 15526137 TI - Acute cholecystitis with a hemocholecyst as an unusual presentation of gallbladder cancer: report of a case. AB - Several atypical presentations of gallbladder carcinoma have been reported, but one of the rarest is intraluminal hemorrhage. We report a case of carcinoma of the gallbladder disclosed by an emergency cholecystectomy, performed for acute cholecystitis caused by a hemocholecyst. The diagnostic approaches and characteristics of a hemocholecyst associated with carcinoma of the gallbladder are discussed. PMID- 15526138 TI - Partial fistulectomy and fistular wall flap for the treatment of high perianal fistulas. AB - Controversy surrounds the management of fistula in ano especially the high type. This study assesses the clinical results of a partial fistulectomy with a closure of the internal opening using a fistular wall flap for the management of high perianal fistulas. This technique was performed in 50 patients with this condition, with the aid of a fistulectomy tube. The clinical outcome was assessed in terms of continence and recurrence for a period of 22 months after operation. The technique is easy to perform, results in less morbidity, and provided successful healing in 49 patients. A recurrent fistula occurred in one patient. No continence disturbance was noted during the follow-up period. This technique is thus considered to be an effective method for managing high perianal fistulas since it was found to demonstrate good results in terms of recurrence and continence. PMID- 15526140 TI - Lack of G protein-coupled sigma receptors in rat brain membranes: receptor mediated high-affinity GTPase activity and [35S]GTPgammaS binding studies. AB - Although sigma (sigma) receptors have been identified as an independent receptor family distinct from opioid and phencyclidine receptors, the physiological roles of these receptors are largely unknown. It is controversial whether there exist metabotropic sigma receptors that are coupled with heterotrimeric G proteins. In the present study, the stimulatory effects of sigma ligands on high-affinity GTPase activity and [35S]GTPgammaS binding were determined in the membranes prepared from rat cerebral cortex, hippocampus, and striatum. In either G protein activation assay, none of the sigma ligands examined had stimulatory effect in any brain regions, except for unambiguous concentration-dependent increase in [35S]GTPgammaS binding by (+)-3-(3-hydroxyphenyl)-N-(1-propyl) piperidine [(+)-3 PPP] in striatal membranes. However, the competition study clearly showed this response was mediated through dopamine D2-like receptors, but not sigma receptors. It is concluded that sigma receptors are not coupled to heterotrimeric G proteins, at least those of Gi/o type. PMID- 15526139 TI - Aminoglutethimide but not spironolactone enhances the anticonvulsant effect of some antiepileptics against amygdala-kindled seizures in rats. AB - Aminoglutethimide (AGLD, an inhibitor of adrenal steroid synthesis) up to 5 mg/kg and spironolactone (SPIR, a mineralocorticosteroid antagonist and a weak antiandrogen) up to 50 mg/kg did not affect any seizure parameter in amygdala kindled rats. AGLD (10 mg/kg) significantly reduced seizure activity in rats of both gender. The combination of AGLD (5 mg/kg) with phenobarbital (PB, applied at its subeffective dose of 15 mg/kg) significantly shortened motor seizure and afterdischarge duration in amygdala-kindled seizures. The combined treatment of AGLD (5 mg/kg) and clonazepam (CLO) at its subeffective dose of 0.01 mg/kg caused significant reduction of the seizure severity, seizure duration and afterdischarge duration. Finally, AGLD (5 mg/kg) proved ineffective upon the action of valproate (VPA) in this model of epilepsy. In contrast to AGLD, SPIR (50 mg/kg) did not affect the action of PB, CLO or VPA against kindled seizures in rats. AGLD did not alter the free plasma levels and brain concentration of PB or CLO, so a pharmacokinetic interaction does not seem probable. Among a variety of chemoconvulsants, bicuculline and N-methyl-D-aspartic acid reversed the effects of AGLD/PB and AGLD/CLO combinations. Aminophylline, kainic acid, strychnine and the glucocorticosteroid (hydrocortisone) were ineffective in this respect. Our data confirm the hypothesis that AGLD-mediated events may play a role in seizure activity and can affect the anticonvulsant activity of some conventional antiepileptic drugs against kindled seizures. Moreover, extrapolation of obtained results to clinical practice may indicate that patients with complex partial seizures may be safely co-medicated with AGLD or SPIR without the risk of worsening of seizure control. PMID- 15526141 TI - Echogenic substantia nigra in patients with orthostatic tremor. AB - OBJECTIVES: Previous studies suggest that the nigrostriatal dopaminergic transmission is impaired in patients with primary orthostatic tremor. METHODS: We used transcranial sonography (TCS) to examine the morphology of the substantia nigra (SN) in four patients with primary orthostatic tremor (OT). RESULTS: TCS revealed an SN echogenicity in all patients, in three patients unilaterally, in one patient bilaterally. CONCLUSIONS: Our data suggest nigrostriatal dopaminergic deficits in OT patients. The exact impact of these dopaminergic deficits on OT generation is unclear. PMID- 15526142 TI - Efficacy and tolerability of a botulinum toxin type A free of complexing proteins (NT 201) compared with commercially available botulinum toxin type A (BOTOX) in healthy volunteers. AB - PURPOSE: This randomized controlled trial was performed to compare the novel botulinum toxin type A free of complexing proteins (NT 201) with the marketed preparation BOTOX degrees regarding efficacy and tolerability. METHODS: Fourteen healthy volunteers received a single intramuscular injection into the extensor digitorum brevis (EDB) muscle of either 4 units NT 201, or 4 units of BOTOX degrees randomised by foot. Compound muscle action potential (CMAP) measurements were recorded for up to 90 days after injection. RESULTS: Both drugs produced a maximum decline between Day 7 and Day 14. At Day 90, administration of both drugs resulted in approximately a 40% CMAP decline as compared to baseline. Duration of paralytic effect was comparable in both groups, at all response thresholds tested. Both drugs were well tolerated. CONCLUSION: The effects of small amounts of NT 201 and BOTOX degrees injected into the EDB muscle are comparable in terms of efficacy, time to onset of action, duration of action, and tolerability. PMID- 15526143 TI - Lack of association between two polymorphisms of brain-derived neurotrophic factor and response to typical neuroleptics. AB - Several studies have connected brain-derived neurotrophic factor (BDNF) with treatment response to neuroleptics. In recent studies, the BDNF expression was reduced by typical neuroleptics. We conducted a retrospective study on 94 patients with schizophrenia and 98 controls. The BDNF G196A and C270T polymorphisms are not associated with treatment response to typical neuroleptics or with age at first hospitalization. Moreover, these polymorphisms of the BDNF gene are not associated with the risk of schizophrenia. PMID- 15526144 TI - RNA synthesis during infection by Hendra virus: an examination by quantitative real-time PCR of RNA accumulation, the effect of ribavirin and the attenuation of transcription. AB - Hendra virus is one of two virus species within the newly-formed genus Henipavirus, subfamily Paramxyovirinae. It is a designated select agent with potential biosecurity threat to both human and animal health. Quantitative real time PCR was used to measure viral RNA synthesis in Vero cells infected by Hendra virus, and to examine the inhibitory effect of ribavirin. It was also used to determine the points of attenuation during transcription of the six viral genes N, P, M, F, G and L by targeting amplicons located towards the 3' end of each gene. Major increases in viral RNA and virus yield occurred between 4 to 8 h and 8 to 10 h post infection, respectively. The effect of ribavirin was examined at a range of concentrations up to 400 microm. At 50 microm, RNA synthesis was reduced 9 fold, and virus yield 58 fold. As expected for a member of the order Mononegavirales, a gradient of transcription was observed in Hendra virus infected cells. There was significant attenuation at the M-F and G-L junctions, more closely resembling Sendai virus (genus Respirovirus) than measles virus (genus Morbillivirus). PMID- 15526145 TI - Partial genome sequence of an apricot isolate of Cherry green ring mottle virus (CGRMV). Annotated sequence record. PMID- 15526146 TI - A novel melon flexivirus transmitted by whitefly. AB - In recent years, a viral disease on melon plants has become a serious problem in Brazil. Symptoms were principally yellowing and mottling on older leaves. Long filamentous virus particles, resembling those of carlaviruses, were seen in symptomatic leaves. In this study, the 3' terminal region of the virus genome isolated from an infected plant, including the last two ORFs, was cloned and sequenced. The sequence comprised a polyadenilated tail and two ORFs, one exhibiting similarity to potexvirus and carlavirus coat protein gene and the second to a carlavirus protein with potential nucleic acid-binding property. The sequence analysis, the genome organization and the particle morphology indicated that the virus could be classified as a novel whitefly-transmitted flexivirus. The name Melon yellowing-associated virus (MYaV) is tentatively suggested for this virus. PMID- 15526147 TI - Modeling of bone at a single lamella level. AB - This paper focuses on the ultrastructure of bone at a single lamella level. At this scale, collagen fibrils reinforced with apatite crystals are aligned preferentially to form a lamella. At the next structural level, such lamella are stacked in different orientations to form either osteons in cortical bone or trabecular pockets in trabecular bone. We use a finite element model, which treats small strain elasticity of a spatially random network of collagen fibrils, and compute anisotropic effective stiffness tensors and deformations of such a single lamella as a function of fibril volume fractions (or porosities), prescribed microgeometries, and fibril geometric and elastic properties. PMID- 15526148 TI - A discrete-time approach to the formulation of constitutive models for viscoelastic soft tissues. AB - This paper presents a novel approach to constitutive modeling of viscoelastic soft tissues. This formulation combines an anisotropic strain energy function, accounting for preferred material directions, to define the elastic stress-strain relationship, and a discrete time black-box dynamic model, borrowed from the theory of system identification, to describe the time-dependent behavior. This discrete time formulation is straightforwardly oriented to the development of a recursive time integration scheme that calculates the current stress state by using strain and stress values stored at a limited number of previous time instants. The viscoelastic model and the numerical procedure are assessed by implementing two numerical examples, the simulation of a uniaxial tensile test and the inflation of a thin tube. Both simulations are performed using parameter values based on previous experiments on preserved bovine pericardium. Parameters are then adjusted to investigate the sensitivity of the model. The hypotheses the model relies upon are discussed and the main limitations are stated. PMID- 15526149 TI - Epigenetic tinkering and evolution: is there any continuity in the role of cytosine methylation from invertebrates to vertebrates? AB - The function of DNA methylation has been investigated in depth in vertebrate and plant genomes, establishing that it is involved in gene silencing and transposon control. Data regarding insect methylation, even if still scanty, apparently argue against evolutionary conservation of DNA methylation functions. Cytosine methylation, therefore, proves to be an epigenetic tool repeatedly used to accomplish different functions in different taxa according to a sort of epigenetic tinkering occurring during evolution. PMID- 15526150 TI - The SARS-CoV S glycoprotein. AB - The severe acute respiratory syndrome-coronavirus (SARS-CoV) spike (S) glycoprotein alone can mediate the membrane fusion required for virus entry and cell fusion. It is also a major immunogen and a target for entry inhibitors. Recent rapid advances in our knowledge of the structure and function of this protein have lead to the development of a number of candidate vaccine immunogens and SARS-CoV entry inhibitors. PMID- 15526151 TI - Biological actions of the oligomers of ribonuclease A. PMID- 15526152 TI - Cys-tRNACys formation and cysteine biosynthesis in methanogenic archaea: two faces of the same problem? AB - Aminoacyl-tRNA (transfer RNA) synthetases are essential components of the cellular translation machinery as they provide the ribosome with aminoacyl-tRNAs. Aminoacyl-tRNA synthesis is generally well understood. However, the mechanism of Cys-tRNACys formation in three methanogenic archaea ( Methanocaldococcus jannaschii, Methanothermobacter thermautotrophicus and Methanopyrus kandleri) is still unknown, since no recognizable gene for a canonical cysteinyl-tRNA synthetase could be identified in the genome sequences of these organisms. Here we review the different routes recently proposed for Cys-tRNACys formation and discuss its possible link with cysteine biosynthesis in these methanogenic archaea. PMID- 15526154 TI - Low-density lipoprotein receptor structure and folding. AB - The endoplasmic reticulum (ER) is a major cellular 'production factory' for many membrane and soluble proteins. A quality control system ensures that only correctly folded and assembled proteins leave the compartment. The low-density lipoprotein receptor (LDLR) is the prototype of a large family of structurally homologous cell surface receptors, which fold in the ER and function as endocytic and signaling receptors in a wide variety of cellular processes. Patients with familial hypercholesterolemia carry single or multiple mutations in their LDLR, which leads to malfunction of the protein, in most patients through misfolding of the receptor. As a result, clearance of cholesterol-rich LDL particles from the circulation decreases, and the elevated blood cholesterol levels cause early onset of atherosclerosis and an increased risk of cardiac disease in these patients. In this review, we will elaborate on the structural aspects of the LDLR and its folding pathway and compare it to other LDLR family members. PMID- 15526153 TI - Evolutionary and functional perspectives of the major histocompatibility complex class I antigen-processing machinery. AB - Major histocompatibility complex (MHC) class I molecules present antigenic peptides to CD8+ T cells, providing the basis for immune recognition of pathogen infected cells. Peptides generated mainly by proteasomes in the cytosol are transported into the lumen of the endoplasmic reticulum by transporters associated with antigen processing (TAP). The maturation of MHC class I molecules is controlled by a number of accessory proteins and chaperones that are to a varying degree dedicated to the assembly of MHC class I. Several newly characterised proteins have been demonstrated to play important roles in this process. This review focuses on the functional relationship and evolutionary history of the antigen-processing machinery (APM) components and MHC class I itself. These are of great interest for further elucidating the origin of the immune system and understanding the mechanisms of antigen presentation and immunology in general. PMID- 15526155 TI - Galactokinase: structure, function and role in type II galactosemia. AB - The conversion of beta- D-galactose to glucose 1-phosphate is accomplished by the action of four enzymes that constitute the Leloir pathway. Galactokinase catalyzes the second step in this pathway, namely the conversion of alpha- D galactose to galactose 1-phosphate. The enzyme has attracted significant research attention because of its important metabolic role, the fact that defects in the human enzyme can result in the diseased state referred to as galactosemia, and most recently for its utilization via 'directed evolution' to create new natural and unnatural sugar 1-phosphates. Additionally, galactokinase-like molecules have been shown to act as sensors for the intracellular concentration of galactose and, under suitable conditions, to function as transcriptional regulators. This review focuses on the recent X-ray crystallographic analyses of galactokinase and places the molecular architecture of this protein in context with the extensive biochemical data that have accumulated over the last 40 years regarding this fascinating small molecule kinase. PMID- 15526156 TI - An update on the biology and physiology of resistin. AB - Resistin is a newly discovered adipocyte hormone. It is related to resistin-like molecules alpha, beta and gamma in structure and function. Resistin is produced by white and brown adipose tissues but has also has been identified in several other tissues, including the hypothalamus, pituitary and adrenal glands, pancreas, gastrointestinal tract, myocytes, spleen, white blood cells and plasma. The tissue level of resistin is decreased by insulin, cytokines such as tumour necrosis factor alpha, endothelin-1 and increased by growth and gonadal hormones, hyperglycaemia, male gender and some proinflammatory cytokines, such as interleukin-6 and lipopolysaccharide. Resistin antagonizes insulin action, and it is downregulated by rosiglitazone and peroxisome proliferator-activated receptor gamma agonists. Since evidence of a direct link between resistin genotype and human diabetes is still weak, more molecular, physiological and clinical studies are needed to determine the role of resistin in the aetiology of type 2 diabetes. PMID- 15526157 TI - Biomimetic actuators: where technology and cell biology merge. AB - The structural and functional analysis of biological macromolecules has reached a level of resolution that allows mechanistic interpretations of molecular action, giving rise to the view of enzymes as molecular machines. This machine analogy is not merely metaphorical, as bio-analogous molecular machines actually are being used as motors in the fields of nanotechnology and robotics. As the borderline between molecular cell biology and technology blurs, developments in the engineering and material sciences become increasingly instructive sources of models and concepts for biologists. In this review, we provide a--necessarily selective--summary of recent progress in the usage of biological and biomimetic materials as actuators in artificial environments, focussing on motors built from DNA, classical cellular motor systems (tubulin/kinesin, actin/myosin), the rotary motor F1F0-ATPase and protein-based 'smart' materials. PMID- 15526158 TI - Nestin expression--a property of multi-lineage progenitor cells? AB - Tissue-specific progenitor cells are characterized by proliferation and differentiation, but, in contrast to embryonic stem (ES) cells, have limited capacities for self-renewal and no tumourigenic potential. These latter traits make progenitor cells an ideal source for regenerative cell therapies. In this review, we describe what is currently known about nestin, an intermediate filament first identified in neuroepithelial stem cells. During embryogenesis, nestin is expressed in migrating and proliferating cells, whereas in adult tissues, nestin is mainly restricted to areas of regeneration. We show that nestin is abundant in ES-derived progenitor cells that have the potential to develop into neuroectodermal, endodermal and mesodermal lineages. Although it remains unclear what factors regulate in vitro and in vivo expression of nestin, we conclude that nestin represents a characteristic marker of multi-lineage progenitor cells and suggest that its presence in cells may indicate multi potentiality and regenerative potential. PMID- 15526159 TI - Immortalization protocols used in cell culture models of human breast morphogenesis. AB - Defining the key players in normal breast differentiation is instrumental to understanding how morphogenesis becomes defective during breast cancer progression. During the past 2 decades much effort has been devoted to the development of technologies for purification and expansion of primary human breast cells in culture and optimizing a relevant microenvironment, which may help to define the niche that regulates breast differentiation and morphogenesis. In contrast to the general property of cancer, normal human cells have a finite lifespan. After a defined number of population doublings, normal cells enter an irreversible proliferation-arrested state referred to as replicative senescence. To overcome this obstacle for continuous long-term studies, replicative senescence can be bypassed by treatment of cells with chemical agents such as benzopyrene, by radiation or by transfection with viral oncogenes or the gene for human telomerase (human telomerase reverse transcriptase, hTERT). A drawback of some of these protocols is a concurrent introduction of chromosomal changes, which sometimes leads to a transformed phenotype and selection of a subpopulation, which may not be representative of the tissue of origin. In recent years, we have sought to establish immortalized primary breast cells, which retain crucial characteristics of their original in situ tissue pattern. This review discusses various approaches to immortalization of breast-derived epithelial and stromal cells and the application of such cell lines for studies on human breast morphogenesis. PMID- 15526160 TI - Signal transduction via the stem cell factor receptor/c-Kit. AB - Together with its ligand, stem cell factor, the receptor tyrosine kinase c-Kit is a key controlling receptor for a number of cell types, including hematopoietic stem cells, mast cells, melanocytes and germ cells. Gain-of-function mutations in c-Kit have been described in a number of human cancers, including testicular germinomas, acute myeloid leukemia and gastrointestinal stromal tumors. Stimulation of c-Kit by its ligand leads to dimerization of receptors, activation of its intrinsic tyrosine kinase activity and phosphorylation of key tyrosine residues within the receptor. These phosphorylated tyrosine residues serve as docking sites for a number of signal transduction molecules containing Src homology 2 domains, which will thereby be recruited to the receptor and activated many times through phosphorylation by the receptor. This review discusses our current knowledge of signal transduction molecules and signal transduction pathways activated by c-Kit and how their activation can be connected to the physiological outcome of c-Kit signaling. PMID- 15526161 TI - Targeted polymeric micelles for delivery of poorly soluble drugs. AB - Polymeric micelles (micelles formed by amphiphilic block copolymers) demonstrate a series of attractive properties as drug carriers, such as high stability both in vitro and in vivo and good biocompatibility, and can be successfully used for the solubilization of various poorly soluble pharmaceuticals. These micelles can also be used as targeted drug delivery systems. The targeting can be achieved via the enhanced permeability and retention effect (into the areas with the compromised vasculature), by making micelles of stimuli-responsive amphiphilic block copolymers, or by attaching specific targeting ligand molecules to the micelle surface. Immunomicelles prepared by coupling monoclonal antibody molecules to p-nitrophenylcarbonyl groups on the water-exposed termini of the micelle corona-forming blocks demonstrate high binding specificity and targetability. Immunomicelles prepared with cancer-specific monoclonal antibody 2C5 specifically bind to different cancer cells in vitro and demonstrate increased therapeutic activity in vivo. This new family of pharmaceutical carriers can be used for the solubilization and targeted delivery of poorly soluble drugs to various pathological sites in the body. PMID- 15526162 TI - The special Sm core structure of the U7 snRNP: far-reaching significance of a small nuclear ribonucleoprotein. AB - The polypeptide composition of the U7 small nuclear ribonucleoprotein (snRNP) involved in histone messenger RNA (mRNA) 3' end formation has recently been elucidated. In contrast to spliceosomal snRNPs, which contain a ring-shaped assembly of seven so-called Sm proteins, in the U7 snRNP the Sm proteins D1 and D2 are replaced by U7-specific Sm-like proteins, Lsm10 and Lsm11. This polypeptide composition and the unusual structure of Lsm11, which plays a role in histone RNA processing, represent new themes in the biology of Sm/Lsm proteins. Moreover this structure has important consequences for snRNP assembly that is mediated by two complexes containing the PRMT5 methyltransferase and the SMN (survival of motor neurons) protein, respectively. Finally, the ability to alter this polypeptide composition by a small mutation in U7 snRNA forms the basis for using modified U7 snRNA derivatives to alter specific pre-mRNA splicing events, thereby opening up a new way for antisense gene therapy. PMID- 15526163 TI - Biochemistry and biology of mammalian DNA methyltransferases. AB - DNA methylation is a stable but not irreversible epigenetic signal that silences gene expression. It has a variety of important functions in mammals, including control of gene expression, cellular differentiation and development, preservation of chromosomal integrity, parental imprinting and X-chromosome inactivation. In addition, it has been implicated in brain function and the development of the immune system. Somatic alterations in genomic methylation patterns contribute to the etiology of human cancers and ageing. It is tightly interwoven with the modification of histone tails and other epigenetic signals. Here we review our current understanding of the molecular enzymology of the mammalian DNA methyltransferases Dnmt1, Dnmt3a, Dnmt3b and Dnmt2 and the roles of the enzymes in the above-mentioned biological processes. PMID- 15526164 TI - Integration target site selection for retroviruses and transposable elements. AB - When a retrovirus infects a cell, its RNA genome is reverse transcribed into a double-stranded DNA, which is then permanently integrated into the host chromosome. Integration is one of the essential steps in the retroviral life cycle. Many transposable elements also move around and integrate into the host genome as part of their life cycle, some through RNA intermediates and some through 'cut and paste' mechanisms. Integration of retroviruses and transposable elements into 'sensitive areas' of the genome can cause irreparable damage. On the other hand, because of their ability to integrate permanently, and the relatively efficient rates of transgenesis, retroviruses and transposable elements are widely used as gene delivery tools in basic research and gene therapy trials. Recent events in gene therapy treatments for X-linked severe combined immunity deficiencies (X-SCID) have highlighted both the promise and some of the risks involved with utilizing retroviruses. Nine of 11 children were successfully treated for X-SCID using a retrovirus carrying the gene mutated in this disease. However, later two of these children developed leukemias because of retroviral integrations in the putative oncogene LMO2 [1]. A third child has also been demonstrated to have an integration in LMO2, but is as of yet nonsymptomatic [2]. It is a bit difficult to explain the high frequency of integrations into the same gene using a random model of retroviral integration, and there has been evidence for decades that retroviral integrations may not be random. But the data were somewhat limited in their power to determine the precise nature of the integration biases. The completion of the human genome sequence coupled with sensitive polymerase chain reaction techniques and an ever-decreasing cost of sequencing has given a powerful new tool to the study of integration site selection. In this review, we describe the findings from several recent global surveys of target site selection by retroviruses and transposable elements, and discuss the possible ramifications of these findings to both mechanisms of action and to the use of these elements as gene therapy vectors. PMID- 15526165 TI - Biological functions of the ING family tumor suppressors. AB - Early studies of the inhibitor of growth 1 ( ING1) gene, the founding member of the ING tumor suppressor family, demonstrated that this gene plays an important role in apoptosis and cellular senescence. Four other related genes have since been identified and found to be involved in various biological activities, including cell cycle arrest, regulation of gene transcription, DNA repair and apoptosis. The biochemical functions of ING proteins as histone acetyltransferases and histone deacetylase co-factors ties this new tumor suppressor family to the regulation of transcription, cell cycle check-points, DNA repair and apoptosis. This review is aimed at summarizing the known biological functions of the ING tumor suppressors and the signalling pathways that they involve. PMID- 15526166 TI - Gelsolin superfamily proteins: key regulators of cellular functions. AB - Cytoskeletal rearrangement occurs in a variety of cellular processes and involves a wide spectrum of proteins. Among these, the gelsolin superfamily proteins control actin organization by severing filaments, capping filament ends and nucleating actin assembly [1]. Gelsolin is the founding member of this family, which now contains at least another six members: villin, adseverin, capG, advillin, supervillin and flightless I. In addition to their respective role in actin filament remodeling, these proteins have some specific and apparently non overlapping particular roles in several cellular processes, including cell motility, control of apoptosis and regulation of phagocytosis (summarized in table 1). Evidence suggests that proteins belonging to the gelsolin superfamily may be involved in other processes, including gene expression regulation. This review will focus on some of the known functions of the gelsolin superfamily proteins, thus providing a basis for reflection on other possible and as yet incompletely understood roles for these proteins. PMID- 15526167 TI - Novel actions of tyrphostin AG 879: inhibition of RAF-1 and HER-2 expression combined with strong antitumoral effects on breast cancer cells. AB - Binding of growth factors to cell surface receptors activates protein tyrosine kinases (PTKs) that initiate cascades of downstream signaling events including the mitogen-activated protein (MAP) kinase cascade. This study reports that the PTK inhibitor AG 879 inhibits proliferation of human breast cancer cells through an effect involving inhibition of MAP kinase activation, but which cannot be explained by effects of AG 879 on its known PTK targets. Instead, AG 879 markedly inhibits expression of the RAF-1 gene, which encodes an upstream MAP kinase kinase kinase. Additionally, expression of HER-2, but not of other genes tested, is inhibited by this compound. These novel effects have to be considered when using AG 879 as a TRK-A and HER-2 inhibitor but may have useful therapeutic implications. PMID- 15526168 TI - Single-copy T-DNAs integrated at different positions in the Arabidopsis genome display uniform and comparable beta-glucuronidase accumulation levels. AB - This study aimed at determining whether transgene expression variability is observed in single-copy T-DNA plants and whether it can be correlated with the T DNA integration position. Among a population of 135 Arabidopsis thaliana transformants, selected on the basis of antibiotic resistance marker expression, 21 single-copy T-DNA transformants were identified and characterized. In 19 of these 21 lines, 35S-beta-glucuronidase transgene expression, measured in two subsequent generations, was similar. This observation means that the intra transformant variability was as high as the inter-transformant variability. Integration into an intergenic or genic region, into an exon or intron, in sense or antisense orientation, did not result in differential transgene expression. Remarkably, single-copy transformants were not always the highest expressers, implying that low transgene expression is not always induced by multicopy transformants. In only 2 of the 21 single-copy plants was the transgene expression more than 20-fold lower. However, characteristics of the insertion position in one of these lines did not differ significantly when compared to high expressing lines. In the remaining line, methylation of the transgene was clearly demonstrated. In conclusion, screening for single-copy T-DNA transformants greatly enriches for stable and high transgene expression, because the integration position is not a major determinant of transgene expression variability in Arabidopsis. PMID- 15526169 TI - Proton motive force mediates a reorientation of the cytosolic domains of the multidrug transporter LmrP. AB - LmrP from Lactococcus lactis is a 45-kDa membrane protein that confers resistance to a wide variety of lipophilic compounds by acting as a proton motive force driven efflux pump. This study shows that both the proton motive force and ligand interaction alter the accessibility of cytosolic tryptophan residues to a hydrophilic quencher. The proton motive force mediates an increase of LmrP accessibility toward the external medium and results in higher drug binding. Residues Asp128 and Asp68, from cytosolic loops, are involved in the proton motive force-mediated accessibility change. Ligand binding does not modify the protein accessibility, but the proton motive force-mediated restructuring is prerequisite for a subsequent accessibility change mediated by ligand binding. Asp142 cooperates with other membrane-embedded carboxylic residues to promote a conformational change that increases LmrP accessibility toward the hydrophilic quencher. This drug binding-mediated reorganization may be related to the transition between the high- and low-affinity drug-binding sites and is crucial for drug release in the extracellular medium. PMID- 15526170 TI - Modulation of SMN nuclear foci and cytoplasmic localization by its C-terminus. AB - The survival of motor neuron ( SMN1) gene product, SMN, is detected both in the cytoplasm and in nuclear gems and cajal bodies. We show here that SMN exon 6 is essential both for formation of its nuclear foci and for its cytoplasmic localization. However, exon 7 inhibits the formation of SMN nuclear foci but promotes SMN cytoplasmic localization. More interestingly, we find that a random C-terminal tag of five or more amino acids downstream of exon 6 is sufficient to inhibit the occurrence of multiple nuclear foci and to promote cytoplasmic localization of SMNDelta7, the primary product of the SMN2 gene. Moreover, SMNDelta7 proteins that bear spinal muscular atrophy mutations in exon 6 either showed defects in nuclear foci formation or enhanced cytoplasmic localization. We conclude that exon 6 and exon 7 synergistically regulate SMN distribution that may require specific exon 6 motifs but is independent of specific sequences in exon 7. PMID- 15526171 TI - Influence of reactive oxygen species production by monoamine oxidase activity on aluminum-induced mitochondrial permeability transition. AB - Treatment of Ca2+-loaded mitochondria with both aluminum and tyramine results in a swelling of higher amplitude than with aluminum alone, while tyramine alone is ineffective. The phenomenon is accompanied by H2O2 production and thiol and pyridine nucleotide oxidation. Cyclosporin A, N-ethylmaleimide or dithioerythritol completely prevent these effects, while catalase exhibits a lower inhibition, pointing to the induction of the permeability transition (MPT) by an oxidative stress. Reactive oxygen species are generated by the interaction of aluminum with the inner membrane and the oxidation of tyramine by monoamine oxidase on the outer membrane. This different localization determines the oxidation of critical thiol groups located on both internal and external sides of pore-forming structures, resulting in MPT induction. The reduced effect by aluminum or the inefficacy by tyramine, when implied alone, can be attributable to the oxidation of thiol groups located only on the internal or external side, respectively. Ultrastructural observations show that aluminum plus tyramine induce the typical configuration of mitochondria that have undergone the MPT. Instead, with aluminum alone, the sensitive subpopulation, although swollen, preserves the outer membrane and shows an apparently orthodox configuration. PMID- 15526175 TI - [Acute cor pulmonale in pulmonary embolism. An important prognostic factor and a critical parameter for the choice of a therapeutic strategy]. AB - Venous thromboembolism remains one of the most frequent and threatening acute cardiovascular syndromes. Its incidence has remained constant over the last several years, both in Europe and the United States, accounting for approximately 1.5 cases per 1000 inhabitants per year. Every year about 100 cases of venous thromboembolism per 100,000 inhabitants are admitted to the hospital, and 10% of these patients die during the hospital stay. Particularly alarming is the fact that in 50-70% of all patients who die from an acute pulmonary embolism the diagnosis is only made after death. These facts emphasize not only the relevance of the problem of "acute pulmonary embolism," but also the need for optimization of current diagnostic and therapeutic strategies. Rapid identification of acute cor pulmonale with echocardiography and cardiac biomarkers has recently proven to be especially helpful in this regard. PMID- 15526176 TI - [When is expensive pharmacotherapy good value for the money?]. AB - Expensive drugs are good value for money if they are effective and safe, and if they have a better cost-effectiveness ratio than the standard therapy. In this article, an overview of commonly used methods, sources and functions of health economic evaluation is presented and illustrated using clinical examples to facilitate interpretation of the health economic information. PMID- 15526177 TI - [Therapeutic management of intra-abdominal and retroperitoneal soft tissue sarcomas]. AB - In this article, we review the diagnostics, therapy, follow-up, and prognosis of intra-abdominal and retroperitoneal soft tissue sarcomas. There are descriptions and discussion of the surgical techniques, advantages and disadvantages of various types and combinations of adjuvant and neoadjuvant therapies, therapy in case of nonresectability, and procedures for desmoid tumors. Further, treatment results, control methods, and studies on new therapeutic approaches are presented. PMID- 15526178 TI - [Bone and soft-tissue tumors of the foot and ankle]. AB - BACKGROUND: This study reports the epidemiology of bone and soft-tissue tumors of the foot and ankle, presents therapy strategies, and evaluates mid-term clinicofunctional outcome after surgery for malignant tumors. METHODS: Two hundred four patients with tumors of the foot and ankle were analyzed (163 benign and 41 malignant). Epidemiology and surgical therapy are reported. RESULTS: The most frequent tumors were exostosis, bone cyst, and osteoid osteoma for benign tumors and metastases and chondrosarcoma and Ewing's sarcoma for malignant tumors. In more than 90% of the benign tumors, local resection could be carried out, whereas in malignant tumors, ablative procedures and arthrodeses were almost as common as limb- and joint-sparing techniques. However, follow-up revealed good functional results and a 5-year survival rate of 84% for patients with primary malignant tumors. CONCLUSION: Tumors of the foot and ankle require a thorough therapeutic strategy. Mid-term functional results and survival rate after surgical treatment are good, although a high percentage of ablative procedures or fusions could not be avoided in our patients. PMID- 15526179 TI - [Ex-vivo gene therapy with BMP-4 for critically sized defects and enhancement of fracture healing in an osteoporotic animal model]. AB - Fractures in osteoporotic bones or segment defects are problematic bone lesions with a reduced biological capability of regeneration. We tested the hypothesis that cell-mediated ex vivo gene therapy to deliver BMP4 can heal critically sized defects and improve bone healing in osteoporotic rats. Primary muscle-derived cells were isolated from the hindlimb muscle of rats and retrovirally transduced to express bone morphogenic protein 4 (BMP4). The bone formation was evaluated following local application of these cells in critically sized defects and in fractures of osteoporotic bones. Radiographic analysis revealed bridging callus formation in a critically sized defect in all specimens using muscle-derived cells expressing BMP4 at 12 weeks. These findings were confirmed by histological evaluation, which revealed callus bone formation with good integration to the distal and proximal bone. Following treatment with muscle-derived cells expressing BMP4, the bone healing process in the osteoporotic bone was improved to the level similar to that of normal bone. The ex vivo gene therapy could be a promising tool for the treatment of osteoporotic fractures and critically sized defects. The reduced number of complications (nonunions, loss of reduction, and fragment dislocation), shortening of hospitalization period, and improvement of bone strength are decisive advocates for this treatment option. PMID- 15526181 TI - [Removal of a thoracic impaling agent without direct observation. The significance of pre-operative diagnosis]. AB - Impalement injuries are uncommon and only occasional reports exist in the literature, resulting in non standardized approaches. Depending on the location, completely different combinations of injuries occur, making every impalement unique. Nevertheless some basic principles for dealing with impalements exist. These principles, and some controversial statements in the literature on the value of preoperative diagnostics, especially CT, are discussed using the example of a spectacular thoracic impalement by a 2 x 2 cm square metal pole. Our deviation from the principal of removing an impaling object only under direct observation in this special case is also discussed. PMID- 15526182 TI - [Differential diagnosis of space demands in the cerebellopontine angle]. AB - Most of the space demands in the cerebellopontine angle lie extra-axially. Important structures run within the cisterns of the cerebellopontine angle, such as the trigeminal, facial and vestibulocochlear nerves as well as the anterior inferior and posterior inferior cerebellar arteries and the veins which lead to the petrosal sinus. The most common space demands are caused by acoustic neuromas, meningeomas, vascular ectasia and aneurysms. Less common are epidermoid and other schwannomas as well as metastases, paragangliomas and arachnoidal cysts. Intra-axial tumours in the area of the cerebellopontine angle include the medulloblastoma, astrocytoma and the ependymoma, which occurs predominantly in children, in addition to the uncommon choroid plexus papilloma. Nearby, there are also space demands around the petrous bone, such as cholesterol granuloma, malignant otitis media, paraganglioma and metastases. For differential diagnosis, an understanding of the space requirements of the tumours in the cerebellopontine angle is needed in addition to knowledge of the anatomical structures. PMID- 15526183 TI - [PET-CT interventional tumour therapy]. AB - Multifocal metastatic spread of solid tumours is often a diagnostic and therapeutic challenge. Those patients often cannot undergo curative surgery. Since several years, various types of interventional tumour therapy have been introduced, which may represent a promising option in those cases. In some cases morphological imaging is not able to assess the viability of a previously treated lesion. The combination of computed tomography and positron emission tomography has the potential to ease planning and guiding of interventional procedures and to improve postinterventional control. Due to its higher sensitivity and specificity, PET/CT enables for a more precise selection of patients for interventional tumor therapy. The success of a particular therapy could be assessed with greater accuracy using PET/CT as compared with the separate modalities. PET/CT was also able to improve planning and guiding interventional therapy. As of yet, no data are available concerning the success rate and long term survival of PET/CT associated interventional procedures. PMID- 15526184 TI - [Therapeutical options for seminomas at clinical stage I-IIA/B]. AB - Seminomas represent the most common histological subgroup of all testicular germ cell tumors. About 75% of all seminomas present as clinical stage I disease at time of initial diagnosis and exhibit a long-term cure rate of 99%. Management strategies maintaining these high cure rates but minimizing the risks need are actively pursued. Currently, three treatment strategies are available for stage I seminomas: surveillance, radiotherapy and chemotherapy. Pathohistological prognostic factors allow an individualized risk-adapted therapeutic approach. Tumor size < or =4 cm and absence of rete testis invasion define a low-risk group with a recurrence rate of 12% being best managed by surveillance. Tumor size >4 cm and presence of rete testis invasion define a high-risk with a 35% risk of relapse, best managed by active therapy. Active treatment either consists of radiation of the ipsilateral paracaval or paraaortic lymph nodes with 20 Gy or of adjuvant chemotherapy with 2 cycles of carboplatin. It is currently unclear if 1 cycle carboplatin is as effective as 2 cycles; if this approach is performed follow-up has to be standardized and a compliance of both patient and physician are mandatory. PMID- 15526185 TI - [Urological education improvement--two paths to a goal]. PMID- 15526186 TI - Post-ICU mortality in critically ill infected patients: an international study. AB - OBJECTIVE: To determine the incidence and risk factors for post-ICU mortality in patients with infection. DESIGN AND SETTING: International observational cohort study including 28 ICUs in eight countries. PATIENTS: All 1,872 patients discharged alive from the ICU over a 1-year period were screened for infection at ICU admission and daily throughout the ICU stay. Outcomes at ICU and hospital discharge were recorded. MEASUREMENTS AND RESULTS: Post-ICU death occurred in 195 (10.4%) patients and was associated in the multivariable analysis with age, chronic respiratory failure, immunosuppression, cirrhosis, Simplified Acute Physiology Score II on the first day with infection, and LOD score at ICU discharge. Post-ICU death was more common among medical patients and patients with hospital-acquired infection or microbiologically documented infection and was less common in patients with pneumonia. CONCLUSIONS: Post-ICU death in patients with infection was within previously reported ranges in overall ICU populations. The main risk factors were patient and infection characteristics, severity at ICU admission, and persistent organ dysfunction at ICU discharge. Further interventions such as further ICU management, discharge to a step-down unit, or follow-up by intensivists on the ward should be evaluated in patients with a high risk of post-ICU mortality. PMID- 15526187 TI - Plasma volume measurement in septic patients using an albumin dilution technique: comparison with the standard radio-labelled albumin method. AB - OBJECTIVE: To investigate a technique using 20% albumin for measurement of plasma volume in critically ill patients. DESIGN AND SETTING: Laboratory and clinical investigation in the adult intensive care unit and anaesthetic laboratories of a university hospital. PATIENTS: 12 patients fulfilling ACCP/SCCM criteria for septic shock. INTERVENTIONS AND MEASUREMENTS: Each patient received (125)I labelled albumin, and the volume of distribution was measured at 1 and 10 min. The accepted standard plasma volume measurement (98% of the 10-min volume of distribution) was calculated. Immediately thereafter 200 ml 20% human albumin was given. Albumin concentrations were measured before and 1 min after this 40-g bolus, and the volume of distribution calculated using a formula that corrected for the 200 ml fluid in which the albumin was dissolved. RESULTS: Plasma volumes measured using the albumin dilution technique at 1 min were smaller than the standard technique by 110+/-280 ml; limits of agreement were from -660 to +440 ml (-16% to +11%). Plasma volumes measured by (125)I-albumin at 1 min were smaller than the standard by 120+/-110 ml; limits of agreement were from -330 to +100 ml (-8 to +2%). CONCLUSIONS: Non-labelled albumin can be used easily and quickly to measure a plasma volume in ICU patients and gives a moderately accurate estimate when compared with the (125)I-labelled albumin methods. The normal isotope method over-estimates plasma volumes in septic patients because excessive transcapillary escape of albumin is inadequately compensated for by the standard correction factor. PMID- 15526188 TI - Will ethical requirements bring critical care research to a halt? PMID- 15526189 TI - Embryonic co-exposure to methoxychlor and Clophen A50 alters sexual behavior in adult male quail. AB - Embryonic exposure to estrogens and estrogenic pollutants is known to demasculinize sexual behavior in adult male Japanese quail. In the present study, we administered the insecticide methoxychlor to quail eggs at a dose of 150 microg/g egg and then studied sexual behavior and other reproductive variables in adult males. In a second experiment we administered the same dose of methoxychlor together with 10 microg/g egg of the commercial polychlorinated biphenyl (PCB) mixture Clophen A50 (CA50) and also CA50 alone. Neither methoxychlor nor CA50 had any significant effects by themselves, but when they were administered together a significant reduction in male sexual behavior was observed. It seems likely that induction of biotransformation enzymes in the embryos by CA50 resulted in increased conversion of methoxychlor to the more estrogenic metabolite 2,2-bis(p hydroxyphenyl)-1,1,1-trichloroethane (HPTE). PMID- 15526190 TI - A new metabolic pathway of arsenite: arsenic-glutathione complexes are substrates for human arsenic methyltransferase Cyt19. AB - The metabolism of arsenic is generally accepted to proceed by repetitive reduction and oxidative methylation; the latter is mediated by arsenic methyltransferase (Cyt19). In human urine, the major metabolites of inorganic arsenicals such as arsenite (iAsIII) and arsenate (iAsV) are monomethylarsonic acid (MMA(V)) and dimethylarsinic acid (DMA(V)). On the other hand, in rat bile, the major metabolites of iAsIII have been reported to be arsenic-glutathione (As GSH) complexes. In the present study we investigate whether these As-GSH complexes are substrates for arsenic methyltransferase by using human recombinant Cyt19. Analyses by high-performance liquid chromatography-inductively coupled plasma mass spectrometry suggested that arsenic triglutathione (ATG) was generated nonenzymatically from iAsIII when GSH was present at concentrations 2 mM or higher. Human recombinant Cyt19 catalyzed transfer of a methyl group from S adenosyl-L-methionine to arsenic and produced monomethyl and dimethyl arsenicals. The methylation of arsenic was catalyzed by Cyt19 only when ATG was present in the reaction mixture. Moreover, monomethylarsonic diglutathione (MADG) was a substrate of Cyt19 for further methylation to dimethylarsinic glutathione (DMAG). On the other hand, monomethylarsonous acid (MMA(III)), a hydrolysis product of MADG, was not methylated to dimethyl arsenical by Cyt19. These results suggest that As-GSH complexes such as ATG and MADG were converted by Cyt19 to MADG and DMAG, respectively. Both MADG and DMAG were unstable in solution when the GSH concentration was lower than 1 mM, and were hydrolyzed and oxidized to MMA(V) and DMA(V), respectively. Metabolism of iAsIII to methylated arsenicals by Cyt19 was via ATG and MADG rather than by oxidative methylation of iAsIII and MMA(III). PMID- 15526191 TI - Oxidation of ethanol to acetaldehyde and free radicals by rat testicular microsomes. AB - A large number of epidemiological studies evidencing that excessive alcohol consumption is associated with impaired testosterone production and testicular atrophy are available in the literature. One hypothesis to explain the deleterious action of alcohol involves the in situ biotransformation to acetaldehyde, but it strongly suggests the need to learn more about the enzymatic processes governing alcohol metabolism to acetaldehyde in different cellular fractions since limited information is available in the literature. In this article we report studies on the metabolic conversion of alcohol to acetaldehyde and to 1-hydroxyethyl radicals in rat testicular microsomal fractions. The oxidation of ethanol to acetaldehyde in rat testes microsomal fraction was mostly of enzymatic nature and strongly dependent on the presence of NADPH and oxygen. Several compounds were able to significantly decrease the production of acetaldehyde: SKF 525A; diethyldithiocarbamate; esculetin; gossypol; curcumin; quercetin; dapsone; and diphenyleneiodonium. Microsomal preparations in the presence of NADPH were also able to produce both hydroxyl and 1-hydroxyethyl free radicals. Their generation was modulated by the presence of diphenyleneiodonium, gossypol, and deferoxamine. Results show that rat microsomal fractions are able to metabolize alcohol to deleterious chemicals, such as acetaldehyde and free radicals, that may be involved in ethanol toxic effects. Enzymes involved could include CYP2E1, P450 reductase, and other enzymes having lipoxygenase- /peroxidase-like behavior. PMID- 15526192 TI - Posture-based or trajectory-based movement planning: a comparison of direct and indirect pointing movements. AB - Various models have been proposed in the literature to explain the control of human arm movements. To make a quantitative comparison between the predictions of various models, we tested subjects for movements to targets on a vertical screen in various conditions. Subjects were asked to move directly from one target to another, or to move by a via-point, at various movement velocities and in a condition with a weight of 0.6 kg attached to the forearm. This set of experimental data was used for comparison with the predictions by various posture based and trajectory-based models on 3-D movement planning and control. Small but significant effects of starting position and path towards the target were found on the torsion of the arm at the end of the movement. No effects of movement velocity and weight attached to the forearm were found. The experimental results differed significantly from the predictions by any of the models considered. Of the models considered, Donders' law best predicts the experimental data. Our data indicate that future tests of models for motor control (1) should compare the predictions of not just one, but several models to a data set, and (2) should include not only planar, but rather 3-D movements in such a comparison. PMID- 15526193 TI - Evidence for entorhinal and parietal cortices involvement in path integration in the rat. AB - Rats with lesions of the entorhinal or parietal cortex were tested in a homing task on a circular platform containing food cups and surrounded by curtains. The animals had to leave a refuge, explore the platform to find a hidden piece of food and carry it back to the refuge. Once the rats were proficient at performing the procedural aspects of the task, they were tested in two successive types of trials in which the food pellet was either always located in the central cup (food at center, "FAC" trials) or placed in a randomly chosen cup (food at random, "FAR" trials). Except in the first FAC trials, all groups displayed similar outward paths in FAC and FAR trials, showing that both types of trials involved equivalent path integration demand. Analysis of the homing accuracy showed that rats with entorhinal cortex or parietal cortex lesions exhibited inaccurate returns to the starting hole, suggesting that these two cortical areas are part of a neural network mediating path integration. PMID- 15526194 TI - Odor-induced changes in taste perception. AB - We investigated odor-induced changes in taste perception (OICTP), by examining the influence of strawberry and soy sauce odors on perceived sweetness (Experiment 1) and saltiness (Experiment 2). We explored whether taste-smell interactions occur at the central level, by delivering odorants (strawberry, soy sauce, odorless water) and tastants (sucrose, sodium chloride) separately, and whether effects of imagined odors are comparable to those of physically presented odors. We found specific taste-smell interactions: sweetness enhancement induced by strawberry odor and saltiness enhancement induced by soy sauce odor. These interactions were elicited with separate delivery of olfactory and gustatory stimuli. Secondly, we found a similar but rather limited effect with the imagined odors: imagined strawberry enhanced perceived sweetness of water solutions, and imagined soy sauce enhanced perceived saltiness of weak sodium chloride solutions. We concluded that OICTP is a centrally mediated phenomenon, and that imagined odors can to some extent induce changes in perceived taste intensity comparable to those elicited by perceived odors. PMID- 15526195 TI - Neonatal bronchial obstruction caused by aspiration of placental tissue. PMID- 15526196 TI - Klebsiella planticola strain DSZ mineralizes simazine: physiological adaptations involved in the process. AB - We examined the ability of a soil bacterium, Klebsiella planticola strain DSZ, to degrade the herbicide simazine (SZ). Strain DSZ is metabolically diverse and grows on a wide range of s-triazine and aromatic compounds. DSZ cells grown in liquid medium with SZ (in 10 mM ethanol) as carbon source mineralized 71.6+/-1.3% of 0.025 mM SZ with a yield of 4.6+/-0.3 microg cell dry weight mmol(-1) carbon. The metabolites produced by DSZ during SZ degradation included ammeline, cyanuric acid, N-formylurea and urea. We studied the physiological adaptations which allow strain DSZ to metabolize SZ. Using scanning electron microscopy, we detected DSZ cells covering the surfaces of SZ crystals when the herbicide was used at high concentrations (0.1 mM). The membrane order observed by FTIR spectroscopy showed membrane activity at low temperature (4 degrees C) to assimilate the herbicide. Membrane fatty acid analysis demonstrated that strain DSZ adapted to grow on SZ by increasing the degree of saturation of membrane lipid fatty acid; and the opposite effect was detected when both SZ and ethanol were used as carbon sources. This confirms the modulator effect of ethanol on membrane fluidity. PMID- 15526198 TI - Stiffness characteristics of hamstring tendon graft fixation methods at the femoral site. AB - In ACL reconstruction, stiffness and strength of a tendon graft complex are important features for knee stability and rehabilitation. The fixation between tendon and bone is known to be one of the weakest components of the graft complex. We examined the tensile load-displacement characteristics of looped semitendinosus tendons in a porcine femoral tunnel. Two groups of six cadaveric semitendinosus tendons and porcine femurs were tested, secured with either an aperture or non-aperture fixation method. Constructs were tested at 1 mm/s until failure in a materials testing machine, which allowed force and displacement data to be recorded. The non-aperture fixation group was significantly less stiff for the first 4 mm of displacement and had significantly higher ultimate failure loads. Provided that adequate ultimate strength can be achieved, stiffness properties of a tendon graft will be improved by using aperture fixation in femoral-site ACL reconstruction. PMID- 15526199 TI - Low back pain after spinal fusion and Harrington instrumentation for idiopathic scoliosis. AB - We reviewed 41 patients with adolescent idiopathic scoliosis treated with spinal fusion and Harrington instrumentation between 1973 and 1992. The mean follow-up was 23 (11-30) years. All patients completed self-administered questionnaires, Oswestry Low Back Pain Disability Score (ODS), Roland Morris score (RLS), and Visual Analog Pain Intensity Scale (VAS). We found a high degree of satisfaction with more than three quarters of the patients in work. The outcome of ODS, RLS, and VAS showed low scores. We found a significant correlation between the scores and the Cobb angle preoperatively as well as at follow-up. The patient-oriented outcome did not correlate with the type of curve, extension of vertebral fusion, tilt angle of the lowest instrumented vertebra, postoperative Cobb angle, loss of correction, or lumbar lordosis. This long-term follow-up of Harrington rod fusion for adolescent idiopathic scoliosis showed no important impairment of health related quality of life. PMID- 15526200 TI - Augmented repair of acute Achilles tendon ruptures using gastrocnemius-soleus fascia. AB - Fifty-four consecutive acute Achilles tendon ruptures were treated with end-to end suture augmented with gastrocnemius fascial flaps. Surgery was performed within 24 h. Mean patient age was 35 (23-57) years, and 46 were men. Mean follow up time was 4.8 (2-8) years. At follow-up, the mean visual analogue scale for pain was 0.49, and the mean AOFAS ankle-hindfoot score was 95 (74-100) points. Fifty-three patients were able to stand on their tiptoes for 30 s and perform repeated toe raises, and 50 patients were able to perform single-limb hopping. Complications included rerupture in one case, deep infection in three cases, delayed wound healing in eight cases, and deep venous thrombosis in one case. Reconstruction of acute ruptures of the Achilles tendon augmented with gastrocnemius-soleus fascial flaps provided a good outcome but was associated with a high complication rate. PMID- 15526201 TI - Evaluation of the cassette dosing approach for assessing the pharmacokinetics of geldanamycin analogues in mice. AB - PURPOSE: There is currently much interest in developing analogues of the benzoquinone ansamycin geldanamycin that may overcome the limitations of 17 (allylamino)-17-demethoxygeldanamycin (17AAG), which is the first known inhibitor of heat shock protein 90 (Hsp90) to enter clinical trials. Studies were performed to assess whether cassette dosing, the coadministration of several compounds to a single animal, is a suitable approach to evaluate the preclinical pharmacokinetics of geldanamycin analogues in high throughput. METHODS: Five geldanamycin analogues (17AAG, NSC 255110, NSC 682300, NSC 683661, NSC 683663) were administered intravenously to mice in combination at 5 mg/kg each and as single agents at 5 mg/kg and 50 mg/kg, or 12.5 mg/kg for NSC 682300. The compounds were also incubated with mouse liver microsomes individually and in combination at 15 microM each. Quantitative analysis was performed by LC/MS/MS. Plasma and tissue pharmacokinetic parameters were evaluated by non-compartmental analysis. In vitro metabolic stability was assessed by monitoring disappearance of the parent compound. RESULTS: Of the compounds that were detectable following individual administration at 5 mg/kg, 17AAG and NSC 683661 exhibited nonlinear pharmacokinetics. In addition, the plasma area under the curve (AUC) and the half life of these compounds was greater following cassette dosing at 5 mg/kg compared to single administration at the same dose. When pharmacokinetic parameters were calculated up to the same time point following cassette and individual administration at the higher dose, three of the compounds displayed non-linear increases in AUC and slower clearances following cassette compared to single compound dosing. When all measurable concentrations at the higher dose were included, the half-life of NSC 683663 was nine-fold longer following individual compared to cassette administration. 17AAG displayed the highest AUC following cassette dosing, whereas NSC 683663 displayed the highest AUC following single compound dosing. Excluding NSC 683663, the rank order from the highest to the lowest AUC was the same; however, NSC 682300, which ranked fifth, was administered at a four-fold lower individual dose than the other compounds. Exposure of the liver and kidneys to the compounds was greater than that of plasma. Despite being administered at a lower dose, NSC 682300 displayed the highest kidney AUC of the five compounds. The same ranking was maintained between cassette and single compound dosing in the kidney. With the exception of NSC 682300, in vitro metabolic stability was predictive of in vivo pharmacokinetics in the plasma and liver. The extent of metabolism of four of the five compounds was lower following microsomal incubation in combination compared to incubation alone, suggestive of likely drug-drug interaction in the cassette. However, for 17AAG this may be partly due to metabolism of NSC 683661 and NSC 683663 to this compound. CONCLUSIONS: Whilst cassette dosing has advantages for use in drug discovery, it is probably unsuitable to evaluate the pharmacokinetics of geldanamycin analogues due to non-linear pharmacokinetics and drug-drug interaction. The issues identified for this compound series should also be considered in assessing the suitability of cassette dosing for other chemotypes. PMID- 15526202 TI - Uptake of the anthracycline pirarubicin into mouse M5076 ovarian sarcoma cells via a sodium-dependent nucleoside transport system. AB - PURPOSE: We have previously demonstrated that the cytotoxicity of anthracyclines, pirarubicin (THP) and doxorubicin (DOX), is partially dominated by their intracellular amounts, which depend on the uptake efficacy of transporter(s). To clarify their transport mechanism, we examined whether or not Na+/nucleoside cotransporter (CNT) is involved in the uptake of THP by M5076 cells. METHODS: Expression of the CNT isoforms was determined by reverse-transcription PCR. We used two cell lines, intact M5076 and CNT2-transfected Cos-7 cells, to characterize the uptake of THP and [3H]uridine. RESULTS: The mRNA for CNT2, but not that for CNT1 or CNT3, was expressed in M5076 cells, and [3H]uridine uptake by the cells required a Na+ gradient as a driving force. THP uptake by M5076 cells depended on a Na+ gradient, and furthermore, formycin B and AZT had cis inhibitory and trans-stimulatory effects on the uptake. The efflux of [3H]uridine from M5076 cells was stimulated by the addition of THP extracellularly, which constituted definite evidence of CNT-mediated uptake of THP. However, THP uptake by CNT2 transfectant was almost the same as that by mock cells, indicating that an unidentified CNT isoform contributes to THP uptake by M5076 cells, this being supported by the differences in transport characteristics of [3H]uridine between M5076 and CNT2-transfected cells. CONCLUSION: THP is partially taken up into M5076 cells via a novel Na+-dependent transport system common to nucleosides. PMID- 15526203 TI - Particular cutaneous side effects with etoposide-containing courses: is VP16 or etoposide phosphate responsible? AB - PURPOSE: Etoposide is commercially available in France in two different pharmaceutical forms: VP16 and its phosphate ester (etoposide phosphate, EP). EP shows better chemical and physical properties, is said to be less toxic but is five times more expensive than VP16. Some criteria were defined for the use of each form in the Paediatric Oncohaematology Department in Hopital Sud in Rennes. As some particular cutaneous side effects were observed during treatment with etoposide-based course in this department, a retrospective study was initiated. The aims of this work were to determine the side effects (especially cutaneous toxicity), whether the pharmaceutical formulation of etoposide had any influence on the toxicity of the drug, and whether the observed side effects resulted from etoposide alone or from particular antineoplastic drug associations. METHODS: Five types of etoposide-containing protocols were chosen: NB 97 and NB 99 (neuroblastoma), FRALLE 93 (acute lymphoid leukaemia), LAME 91 (acute myeloid leukaemia), OS 94 (osteosarcoma), Ewing 97 and Euro-Ewing 99 (Ewing sarcoma). The medical files of 36 children (88 EP courses, 25 VP16 courses) included in these protocols were analysed on the basis that if a child showed a side effect during a course, the child had to have recovered from that side effect before the beginning of the next course. RESULTS: Apart from classical side effects (haematological and digestive toxicities etc.), two particular cutaneous side effects were observed: (1) palmar-plantar eruptions and nail inflammations, and (2) irritation of the anal area and anal fissures. Those side effects were observed with three of the studied protocols: NB 97, OS 94 and Ewing sarcoma treatments. CONCLUSIONS: No striking differences in toxicity appeared between the two etoposide formulations, but this retrospective study seemed to confirm the appearance of particular cutaneous and anal side effects especially with two associations: (1) etoposide-ifosfamide (OS 94 and Ewing 97), and (2) etoposide ifosfamide-Adriamycin-vincristine (VIDE course of the Euro-Ewing 99 protocol). PMID- 15526204 TI - Structural features and transcript-editing analysis of sugarcane (Saccharum officinarum L.) chloroplast genome. AB - The complete nucleotide sequence of the chloroplast genome of sugarcane (Saccharum officinarum) was determined. It consists of 141,182 base-pairs (bp), containing a pair of inverted repeat regions (IR(A), IR(B)) of 22,794 bp each. The IR(A) and IR(B) sequences separate a small single copy region (12,546 bp) and a large single copy (83,048 bp) region. The gene content and relative arrangement of the 116 identified genes (82 peptide-encoding genes, four ribosomal RNA genes, 30 tRNA genes), with the 16 ycf genes, are highly similar to maize. Editing events, defined as C-to-U transitions in the mRNA sequences, were comparable with those observed in maize, rice and wheat. The conservation of gene organization and mRNA editing suggests a common ancestor for the sugarcane and maize plastomes. These data provide the basis for functional analysis of plastid genes and plastid metabolism within the Poaceae. The sugarcane chloroplast DNA sequence is available at GenBank under accession NC005878. PMID- 15526205 TI - Characterization of the Arxula adeninivorans AHOG1 gene and the encoded mitogen activated protein kinase. AB - Arxula adeninivorans is an osmo-resistant yeast species that can tolerate high levels of osmolytes like NaCl, PEG400 and ethylene glycol. As in other yeast species, this tolerance is elicited by components of the high osmolarity glycerol (HOG) response pathway. In the present study, we isolated and characterized as a key component of this pathway the A. adeninivorans AHOG1 gene encoding the mitogen-activated protein (MAP) kinase Ahog1p, an enzyme of 45.9 kDa. The gene includes a coding sequence of 1,203 bp disrupted by a 57-bp intron. The identity of the gene was confirmed by complementation of a hog1 mutation in a Saccharomyces cerevisiae mutant strain and the high degree of homology of the derived amino acid sequence with that of MAP kinases from other yeasts and fungi. Under stress-free conditions, the inactive Ahoglp is present in low levels. When exposed to osmotic stress, Ahoglp is rendered active by phosphorylation. In addition, AHOG1 expression is increased. Assessment of the AHOG1 promoter activity with a lacZ reporter gene confirmed its inducibility by osmolytes, a characteristic not observed in homologous HOG1 genes of other yeast species. This specific property could account for the fast adaptation and high osmo-resistance encountered in this species. PMID- 15526206 TI - The gene for the heat-shock protein HSP100 is induced by blue light and heat shock in the fungus Phycomyces blakesleeanus. AB - We cloned and sequenced the Phycomyces hspA gene. The hspA gene product is a 901 amino-acid protein member of the clpB/HSP100 family. HSP100 proteins are ATPases involved in high-temperature tolerance, proteolysis, and protein disaggregation. Phycomyces HSP100 is composed of a domain presumably involved in protein-protein interactions and two ATP-binding domains. The hspA promoter contains three heat shock elements that are presumably involved in the activation of hspA after heat shock. In addition, four short sequences are present in the hspA promoter and in the promoter of the photoinducible genes carB and carRA; and these are candidates as binding sites for light-regulated transcription factors. Blue light can increase transcription of the hspA gene 10-fold, with a threshold of 1 J/m2. The threshold for hspA photoactivation is 10(4) times higher than the thresholds for blue-light regulation of sporangiophore development and photocarotenogenesis, which suggests that there are differences in the photosensory systems for gene photoactivation and mycelial photoresponses. A heat-shock of 30 min at 34 degrees C or 42 degrees C increased hspA gene activity 160-fold. The differences in maximum hspA gene transcription by blue light and heatshock suggest the presence of different regulatory mechanisms. PMID- 15526207 TI - Detection of pulmonary nodules at multirow-detector CT: effectiveness of double reading to improve sensitivity at standard-dose and low-dose chest CT. AB - The purpose of this study was to assess the effectiveness of double reading to increase the sensitivity of lung nodule detection at standard-dose (SDCT) and low dose multirow-detector CT (LDCT). SDCT (100 mAs effective tube current) and LDCT (20 mAs) of nine patients with pulmonary metastases were obtained within 5 min using four-row detector CT. Softcopy images reconstructed with 5-mm slice thickness were read by three radiologists independently. Images with 1.25-mm slice thickness served as the gold standard. Sensitivity was assessed for single readers and combinations. The effectiveness of double reading was expressed as the increase of sensitivity. Average sensitivity for detection of 390 nodules (size 3.9+/-3.2 mm) for single readers was 0.63 (SDCT) and 0.64 (LDCT). Double reading significantly increased sensitivity to 0.74 and 0.79, respectively. No significant difference between sensitivity at SDCT and LDCT was observed. The percentage of nodules detected by all three readers concordantly was 52% for SDCT and 47% for LDCT. Although double reading increased the detection rate of pulmonary nodules from 63% to 74-79%, a considerable proportion of nodules remained undetected. No difference between sensitivities at LDCT and SDCT for detection of small nodules was observed. PMID- 15526208 TI - Meal ingestion provokes entry of lipoproteins containing fat from the previous meal: possible metabolic implications. AB - BACKGROUND: Prolonged and exaggerated postprandial plasma triacylglycerol (TAG) concentrations are considered as an independent risk factor for coronary artery disease. Western populations eat many meals at regular intervals, and can be in a postprandial state for at least 17 h of a 24 h period. After consuming 2 meals an early plasma TAG peak has been observed after the second meal, the origin of which is unclear. AIM OF THE STUDY: To test the hypothesis that the early TAG peak observed following sequential meals was of intestinal origin and represented fat derived from the previous meal. METHODS: Postprandial plasma lipaemic responses of 17 healthy postmenopausal women were studied by giving a test breakfast followed by a lunch. Watermiscible retinyl palmitate (RP) was added to the breakfast, but not the lunch test meal. Plasma TAG, retinyl esters (RE) and apo B-48 were determined for a 10 h period following breakfast. RESULTS: In response to the test meals, RE, apo B-48 and TAG showed multiple peaks. Despite omission of RP from the lunch, RE showed an early peak response after ingestion of lunch in 15 of 17 subjects. The peak response after lunch of all three markers appeared significantly earlier compared with their respective peak responses after the breakfast (P < 0.0001). The area of RE response after lunch was significantly correlated with the RE lipaemic response to the breakfast (r = 0.67; P < 0.004) and to the fasting TAG concentration (r = 0.48; P < 0.05). CONCLUSIONS: Since the lunch did not contain RP, the distinctive second influx of RE after lunch was believed to have originated from the breakfast. This, together with the fact that all three markers showed an earlier response to the lunch than the breakfast, supports the view that ingestion of a second meal provokes entry of fat from the previous meal, from an as yet unidentified site (gut, enterocytes, lymph). The results indicate that the degree of TAG "storage" from previous meals might be a function of TAG tolerance and provide a possible site of regulation of the entry of fat into the systemic circulation. PMID- 15526209 TI - In vitro fermentation characteristics of a mixture of Raftilose and guar gum by human faecal bacteria. AB - BACKGROUND: The therapeutic effects of indigestible carbohydrates in the human colon are well known. Most fermentation studies have examined only single carbohydrates. Considering the idiosyncratic actions of individual carbohydrates and the dose required, it is unlikely that any one carbohydrate will provide ideal physiological actions without having some undesirable effects. Little is known of how mixing non-digestible carbohydrates affects their fermentation. This necessitates fermentation studies using different carbohydrates in mixtures. AIM OF THE STUDY: The aim of this study was to test the effect of mixing Raftilose (R) and guar gum (G) on short chain fatty acid (SCFA) production in in vitro cultures of human faecal bacteria. METHOD: The fermentation of the individual carbohydrates (10 mg/ml) was compared with that of a 50:50 mixture in anaerobic in vitro cultures of human faeces. RESULTS: Cultures of R/G mixtures produced significantly more n-butyrate than 100 mg G alone at 8 and 24 hours (p < 0.02). There was no significant difference in the production of n-butyrate between cultures of 100 mg R and the R/G mixture at 8 and 24 hours. R (100 mg) produced a propionic/butyric (p/b) acid ratio of 1.18 compared with 3.88 for 100 mg G, whereas R/G mixture produced a p/b ratio of 2.01. CONCLUSION: The fermentation of R/G mixture was different compared with 100 mg of R or G alone. There was no loss of n-butyrate in the culture containing R/G (50 mg of each) mixture compared with the 100 mg R culture. PMID- 15526210 TI - Ovarian teratoma with diffused peritoneal reactions mimicking advanced ovarian malignancy. AB - BACKGROUND: Benign cystic teratomas are one of the most common benign ovarian neoplasms. Although its rupture is rare, once occurred it can cause complications such as granulomatous peritonitis, mimicking metastatic ovarian malignancy. CASE: A 39-year-old woman, Para 0-0-0-0, presented to the hospital with rapid abdominal distention for 3 months. Her physical examination and ultrasonographic findings led to a diagnosis of advanced stage ovarian carcinoma. An exploratory laparotomy was performed and the operative impression was that of stage III ovarian carcinoma. Total hysterectomy with bilateral salpingo-oophorectomy and surgical staging were done. The postoperative pathology revealed a benign cystic teratoma of right ovary with chronic granulomatous peritonitis. She was well at discharge and at her 1-year follow-up. CONCLUSION: Although ruptured a benign cystic teratoma is rare, it can cause granulomatous peritonitis, the clinical findings of which mimic advanced stage ovarian carcinoma. This warrants physicians to be aware of and intraoperative frozen section should be used, its correct management will provide a good outcome with less complication. PMID- 15526211 TI - Forensic application of VEGF expression to skin wound age determination. AB - An immunohistochemical study combined with morphometry was carried out to examine the time-dependent expression of vascular endothelial growth factor (VEGF) using 53 human skin wounds with different wound ages (groups I: 0-12 h, II: 1-4 days, III: 7-14 days and IV: 17-21 days). In the human wound specimens aged 4-12 h, neutrophils recruited at the wound showed no positive signals for VEGF. With an increase in wound ages of > or =7 days, granulation tissue and angiogenesis were observed, with the migration of macrophages and fibroblasts of which the cytoplasm expressed VEGF-positive reactions. Morphometrically, the average VEGF positive ratio was highest in group III, followed by that of group IV. In groups III and IV, 13 out of 26 wound samples had VEGF-positive ratios of more than 50%. However, all of the wound samples in groups I and II showed VEGF-positive ratios of less than 50%. With regard to the practical applicability and forensic validity, these observations suggest that a VEGF-positive ratio of more than 50% possibly indicates a wound age of 7 days or more. PMID- 15526212 TI - Development of a psilocin immunoassay for serum and blood samples. AB - After the immunisation of rabbits with a psilocin-specific immunogen, polyclonal antisera were obtained. With these antisera a competitive, heterogeneous radioimmunoassay for the detection of psilocin was developed. As tracer a derivative of psilocin was synthesised, which contained a tritiated CH(3) group. The antisera showed a specific reaction with psilocin. The cross-reactivity of structurally related endogenous substances like serotonin, tryptophan and tyrosine was below 0.01%. Also common drugs of abuse (Delta(9) tetrahydrocannabinol, cocaine, morphine, amphetamine) showed negligible cross reactivity (0.01-2%). Only tricyclic neuroleptics with a (dimethylamino)ethyl side-chain showed some cross-reactivity (20%). Spiked serum and blood samples were analysed with this new immunoassay and the results obtained were compared with the values measured with a validated GC-MS method. PMID- 15526213 TI - Mortality and cancer incidence in a Swedish art glassworks--an updated cohort study. AB - AIM: To study mortality and cancer incidence, in a Swedish art glassworks producing both heavy and semi-crystal glassware, in an extended cohort of workers over a long time period during which some preventive actions had taken place. METHODS: In the updated study, 1,229 men and women were eligible as cohort members during the period 1964-1997. The observed number of cases was compared with expected numbers, as calculated from cause-, age-, gender- and calendar year specific national rates for mortality and cancer incidence. RESULTS: Among men only, a significant risk was seen for cancer incidence in the colon and rectum [standardised incidence ratio (SIR) 1.92, 95% confidence interval (CI) 1.05-3.23; 14 cases] and increased, but statistically non-significant, risks were also seen for male cases of tumours in the liver/bile ducts and brain. Among women, statistically non-significant risks were seen for tumours in the liver/bile ducts and in the lymphatic and haematopoietic systems. No increased risk for cancer of the lung was found in this updated study. The risk for cancer in the colon/rectum was slightly increased in all work categories, and the increase was statistically significant among male and female unspecified glassworkers (SIR 3.13, 95% CI 1.35 6.16; five male and three female cases). A statistically significantly increased risk for cancer in the liver/bile ducts was seen among refinement workers (SIR 3.96, 95% CI 1.07-10.14; two male and two female cases). CONCLUSION: Most of the causes of death associated with an elevated standardised mortality ratio (SMR) in the 1985 cohort resulted in lower SMRs in this updated cohort, maybe as a consequence of preventive actions taken at the glassworks. On the other hand, the risk for cancers in the digestive system seems to remain, perhaps due to past asbestos exposure or inhalation/digestion of larger particles in the ambient air. PMID- 15526214 TI - Lipoxygenase-mediated metabolism of storage lipids in germinating sunflower cotyledons and beta-oxidation of (9Z,11E,13S)-13-hydroxy-octadeca-9,11-dienoic acid by the cotyledonary glyoxysomes. AB - During the early stages of germination, a lipid-body lipoxygenase is expressed in the cotyledons of sunflowers (Helianthus annuus L.). In order to obtain evidence for the in vivo activity of this enzyme during germination, we analyzed the lipoxygenase-dependent metabolism of polyunsaturated fatty acids esterified in the storage lipids. For this purpose, lipid bodies were isolated from etiolated sunflower cotyledons at different stages of germination, and the storage triacylglycerols were analyzed for oxygenated derivatives. During the time course of germination the amount of oxygenated storage lipids was strongly augmented, and we detected triacylglycerols containing one, two or three residues of (9Z,11E,13S)-13-hydro(pero)xy-octadeca-9,11-dienoic acid. Glyoxysomes from etiolated sunflower cotyledons converted (9Z,11E,13S)-13-hydroxy-octadeca-9,11 dienoic acid to (9Z,11E)-13-oxo-octadeca-9,11-dienoic acid via an NADH-dependent dehydrogenase reaction. Both oxygenated fatty acid derivatives were activated to the corresponding CoA esters and subsequently metabolized to compounds of shorter chain length. Cofactor requirement and formation of acetyl-CoA indicate degradation via beta-oxidation. However, beta-oxidation only proceeded for two consecutive cycles, leading to accumulation of a medium-chain metabolite carrying an oxo group at C-9, equivalent to C-13 of the parent (9Z,11E,13S)-13-hydroxy octadeca-9,11-dienoic acid. Short-chain beta-oxidation intermediates were not detected during incubation. Similar results were obtained when 13-hydroxy octadecanoic acid was used as beta-oxidation substrate. On the other hand, the degradation of (9Z,11E)-octadeca-9,11-dienoic acid was accompanied by the appearance of short-chain beta-oxidation intermediates in the reaction mixture. The results suggest that the hydroxyl/oxo group at C-13 of lipoxygenase-derived fatty acids forms a barrier to continuous beta-oxidation by glyoxysomes. PMID- 15526215 TI - Ci-POU-IV expression identifies PNS neurons in embryos and larvae of the ascidian Ciona intestinalis. AB - Several lines of evidence suggest that members of the POU domain gene family may regulate invertebrate and vertebrate neurogenesis. In particular, POU IV genes appear to be neural genes involved in differentiation of sensory neurons, as demonstrated in mollusc, Drosophila, Caenorhabditis elegans and vertebrates. In the present work, we describe the developmental expression of a homologue of POU IV genes, Ci-POU-IV, in the ascidian Ciona intestinalis. Ci-POU-IV is expressed in the precursor cells of the neural system during development and in the neural system of the larva. In particular, transcripts are prevalent in the peripheral nervous system (PNS), with expression in the central nervous system (CNS) restricted to the posterior sensory vesicle. Therefore, the evolution of a complex sensory system seems to be under the control of a common genetic mechanism. PMID- 15526216 TI - Thalamic projections to the auditory cortex in the rufous horseshoe bat (Rhinolophus rouxi). II. Dorsal fields. AB - In this study, we analyzed the thalamic connections to the parietal or dorsal auditory cortical fields of the horseshoe bat, Rhinolophus rouxi. The data of the present study were collected as part of a combined investigation of physiologic properties, neuroarchitecture, and chemoarchitecture as well as connectivity of cortical fields in Rhinolophus, in order to establish a neuroanatomically and functionally coherent view of the auditory cortex. Horseradish peroxidase or wheat-germ-agglutinated horseradish peroxidase deposits were made into cortical fields after mapping response properties. The dorsal fields of the auditory cortex span nearly the entire parietal region and comprise more than half of the non-primary auditory cortex. In contrast to the temporal fields of the auditory cortex, which receive input mainly from the ventral medial geniculate body (or "main sensory nucleus"), the dorsal fields of the auditory cortex receive strong input from the "associated nuclei" of the medial geniculate body, especially from the anterior dorsal nucleus of the medial geniculate body. The anterior dorsal nucleus is as significant for the dorsal fields of the auditory cortex as the ventral nucleus of the medial geniculate body is for the temporal fields of the auditory cortex. Additionally, the multisensory nuclei of the medial geniculate body provide a large share of the total input to the nonprimary fields of the auditory cortex. Comparing the organization of thalamic auditory cortical afferents in Rhinolophus with other species demonstrates the strong organizational similarity of this bat's auditory cortex with that of other mammals, including primates, and provides further evidence that the bat is a relevant and valuable model for studying mammalian auditory function. PMID- 15526217 TI - Cytoarchitecture of the medial geniculate body and thalamic projections to the auditory cortex in the rufous horseshoe bat (Rhinolophus rouxi). I. Temporal fields. AB - The auditory cortex in echolocating bats is one of the best studied in mammals, yet the projections of the thalamus to the different auditory cortical fields have not been systematically analyzed in any bat species. The data of the present study were collected as part of a combined investigation of physiological properties, neuroarchitecture, and chemoarchitecture as well as connectivity of cortical fields in Rhinolophus in order to establish a neuroanatomically and functionally coherent view of the auditory cortex in the horseshoe bat. This paper first describes the neuroanatomic parcellation of the medial geniculate body and then concentrates on the afferent thalamic connections with auditory cortical fields of the temporal region. Deposits of horseradish peroxidase and wheatgerm-agglutinated horseradish peroxidase were made into neurophysiologically characterized locations of temporal auditory cortical fields; i.e., the tonotopically organized primary auditory cortex, a ventral field, and a temporal subdivision of a posterior dorsal field. A clear topographic relationship between thalamic subdivisions and specific cortical areas is demonstrated. The primary auditory cortex receives topographically organized input from the central ventral medial geniculate body. The projection patterns to the temporal subdivision of the posterior dorsal field suggest that it is a "core" field, similar to the posterior fields in the cat. Projections to the ventral field arise primarily from border regions of the ventral medial geniculate body. On the whole, the organization of the medial geniculate body projections to the temporal auditory cortex is quite similar to that described in other mammals, including cat and monkey. PMID- 15526218 TI - Non-recurrent 17p11.2 deletions are generated by homologous and non-homologous mechanisms. AB - Several recurrent common chromosomal deletion and duplication breakpoints have been localized to large, highly homologous, low-copy repeats (LCRs). The mechanism responsible for these rearrangements, viz., non-allelic homologous recombination between LCR copies, has been well established. However, fewer studies have examined the mechanisms responsible for non-recurrent rearrangements with non-homologous breakpoint regions. Here, we have analyzed four uncommon deletions of 17p11.2, involving the Smith-Magenis syndrome region. Using somatic cell hybrid lines created from patient lymphoblasts, we have utilized a strategy based on the polymerase chain reaction to refine the deletion breakpoints and to obtain sequence data at the deletion junction. Our analyses have revealed that two of the four deletions are a product of Alu/Alu recombination, whereas the remaining two deletions result from a non-homologous end-joining mechanism. Of the breakpoints studied, three of eight are located in LCRs, and five of eight are within repetitive elements, including Alu and MER5B sequences. These findings suggest that higher-order genomic architecture, such as LCRs, and smaller repetitive sequences, such as Alu elements, can mediate chromosomal deletions via homologous and non-homologous mechanisms. These data further implicate homologous recombination as the predominant mechanism of deletion formation in this genomic interval. PMID- 15526219 TI - Anatomical study of the paraspinal approach to the lumbar spine. AB - The original description of the paraspinal posterior approach to the lumbar spine was for spinal fusion, especially regarding lumbosacral spondylolisthesis treatment. In spite of the technical details described by Wiltse, exact location of the area where the sacrospinalis muscle has to be split remains somewhat unclear. The goal of this study was to provide topographic landmarks to facilitate this surgical approach. Thirty cadavers were dissected in order to precisely describe the anatomy of the trans-muscular paraspinal approach. The level of the natural cleavage plane between the multifidus and the longissimus part of the sacrospinalis muscle was noted and measurements were done between this level and the midline at the level of the spinous process of L4. A natural cleavage plane between the multifidus and the longissimus part of the sacrospinalis muscle was present in all cases. There was a fibrous separation between the two muscular parts in 55 out of 60 cases. The mean distance between the level of the cleavage plane and the midline was 4 cm (2.4-5.5 cm). In all cases, small arteries and veins were present, precisely at the level of the cleavage plane. We found it possible to easily localize the anatomical cleavage plane between the multifidus part and the longissimus part of the sacrospinalis muscle. First the superficial muscular fascia is opened near the midline, exposing the posterior aspect of the sacrospinalis muscle. Then, the location of the muscular cleft can be found by identifying the perforating vessels leaving the anatomical inter-muscular space. PMID- 15526220 TI - The role of closed-suction drainage in preventing epidural fibrosis and its correlation with a new grading system of epidural fibrosis on the basis of MRI. AB - In this study we aimed to evaluate the role of closed-suction drainage on the extent of epidural fibrosis (EF) after lumbar disc surgery and to define a new grading system of epidural fibrosis in these patients, based on magnetic resonance imaging. Seventy-nine patients (34 women, 45 men) with a unilateral, single-level lumbar disc herniation were included in this study. Forty-one patients in whom closed-suction drainage was implanted were compared with 38 patients in whom the drain was not implanted. We have used a new grading system for the extent of epidural fibrosis, on the basis of follow-up magnetic resonance imaging findings. Pain intensity was evaluated by visual analog scale (VAS), and the patients' function and working ability were measured according to the Prolo functional-economic scale. We conclude that, in patients operated on for unilateral, single-level lumbar disc hernias, implantation of closed-suction drainage into the operation site results in less formation of EF radiologically and yields better clinical outcome. PMID- 15526221 TI - Surface-based registration accuracy of CT-based image-guided spine surgery. AB - Registration is a critical and important process in maintaining the accuracy of CT-based image-guided surgery. The aim of this study was to evaluate the effects of the area of intraoperative data sampling and number of sampling points on the accuracy of surface-based registration in a CT-based spinal-navigation system, using an optical three-dimensional localizer. A cadaveric dry-bone phantom of the lumbar spine was used. To evaluate registration accuracy, three alumina ceramic balls were attached to the anterior and lateral aspects of the vertebral body. CT images of the phantom were obtained (1-mm slice thickness, at1-mm intervals) using a helical CT scanner. Twenty surface points were digitized from five zones defined on the basis of anatomical classification on the posterior aspects of the target vertebra. A total of 20 sets of sampling data were obtained. Evaluation of registration accuracy accounted for positional and rotational errors. Of the five zones, the area that was the largest and easiest to expose surgically and to digitize surface points was the lamina. The lamina was defined as standard zone. On this zone, the effect of the number of sampling points on the positional and rotational accuracy of registration was evaluated. And the effects of the additional area selected for intraoperative data sampling on the registration accuracy were evaluated. Using 20 surface points on the posterior side of the lamina, positional error was 0.96 mm +/- 0.24 mm root-mean-square (RMS) and rotational error was 0.91 degrees +/- 0.38 degrees RMS. The use of 20 surface points on the lamina usually allows surgeons to carry out sufficiently accurate registration to conduct computer-aided spine surgery. In the case of severe spondylosis, however, it might be difficult to digitize the surface points from the lamina, due to a hypertrophic facet joint or the deformity of the lamina and noisy sampling data. In such cases, registration accuracy can be improved by combining use of the 20 surface points on the lamina with surface points on other zones, such as on the both sides of the spinous process. PMID- 15526222 TI - Measurement of angular and linear segmental lumbar spine flexion-extension motion by means of image registration. AB - BACKGROUND: The presently available method of measuring segmental lumbar spine mobility by means of superimposition of lumbar spine radiographs in flexion and extension lacks precision due to differences in the cortical outline of the vertebral bodies in flexed and extended position. The introduction of digital image processing has opened the possibility of computerised superimposition ('matching') of digital vertebral body images by means of image registration. Theoretically this technique allows more accurate image matching and, consequently, greater precision of measurement because the whole vertebral body image (not only its cortical outline) can be chosen as region of interest, with registration of all available digital information within this region. METHODS: To check accuracy and convenience of the new method, two computer program experts performed five image registration measurements of the five lumbar motion segments in five consecutive flexion-extension studies of old lumbar fracture, spondylolytic spondylolisthesis and degenerative anterolisthesis. For comparison an experienced radiologist performed the same repeated measurements with the manual superimposition method. RESULTS: Measurement error of the image registration method proved to be significantly smaller than that of the manual superimposition method. There was no overlap between the 95% confidence intervals of the mean standard deviations of experts A and B using the image registration method and the 95% confidence interval of the mean standard deviations of the experienced radiologist using the manual superimposition method. Besides, the image registration method proved to be more convenient because the whole procedure from import of the image data to display of the measurement outcomes lasted 2-3 min compared to 3-6 min for the superimposition method. PMID- 15526223 TI - Intramedullary spinal cord metastases: diagnosis and treatment - an illustrated review. AB - BACKGROUND: Intramedullary spinal cord metastases (ISCM) are rare but, with increasing use of magnetic resonance imaging (MRI) are being encountered with increasing frequency. Optimum treatment remains controversial. On the basis of a review of previous reports and experience with a patient with an ISCM from a large cell lung cancer, we propose practical diagnostic and therapeutic approaches. FINDINGS: We found 284 patients who had an Intramedullary spinal cord metastasis reported in English literature up to February 2004. 32 had been treated surgically. The mean survival in these patients was two times longer than in those treated by a conservative approach. Improvement and prolonged survival occurred in patient we treated by microsurgical dissection of the metastasis. CONCLUSION: Early diagnosis and early surgical resection can result in improvement in neurological deficits and in the quality of life of patients with a Intramedullary spinal cord metastasis. PMID- 15526227 TI - Dairy cream as a phantom material for biexponential diffusion decay. AB - Commercially available aliquots of dairy cream are shown to have diffusion decay curves characterized by biexponential functions when studied over a wide range of b-factors. The fast and slow diffusion components responsible for the biexponential decay are attributed to water and lipid protons, respectively. The fast diffusion coefficient and relative fast and slow diffusion component fractions obtained from biexponential fits of cream phantoms over a wide range of b-factors up to 3,000 s/mm2 are similar to those obtained previously for brain. The slow diffusion coefficient from lipid protons is smaller than that found in the brain. Overall, however, the results suggest that dairy cream can serve as a widely available phantom material for testing software and hardware components designed to perform quantitative, biexponential diffusion decay studies. PMID- 15526228 TI - [Urinary stress incontinence in women]. AB - The integral theory of Petros and Ulmsten has profoundly changed our understanding of the female pelvic floor. Anatomic laxity of the vaginal wall caused by pelvic floor defects induced at different damage zones is frequently not only responsible for stress urinary incontinence but also for pollakisuria, urgency, post-void residual and pelvic pain. A number of minimally invasive techniques have been developed to correct these defects. Applying a tension-free polypropylene tape around the mid-urethra has become an established method to correct the anterior ligaments. The infra-coccygeal sacropexy can achieve dorsal stabilization of the vaginal wall. Currently, polypropylene meshes are increasingly used for repairing supporting pelvic fasciae. The most recommended conservative methods are exercises to strengthen the pelvic floor muscles. Duloxetine increases the rhabdosphincter contractility during the filling phases, but not during voiding, and therefore is a promising drug for clinical use. PMID- 15526229 TI - [Proteasome inhibitors: induction of apoptosis as new therapeutic option in prostate cancer]. AB - New perspectives in prostate cancer genesis and putative clinical management have emerged in recent years . Apoptosis plays a major role in this environment. Proteasome inhibitors block the action of a multicatalytic proteinase complex involved in the degradation of intracellular proteins, particularly with regard to cell cycle regulation and apoptosis. Numerous in vitro studies have demonstrated the ability of these compounds to induce apoptosis and enhance the activity of conventional tumoricidal agents in many cancer cell types, including prostate cancer cells. They point out the use of these potent inhibitors as a new potential molecular approach to the therapeutic management of prostate cancer. Furthermore, the action of proteasome inhibitors has been tested in animal models and in patients with hormone refractory prostate cancer, resulting in both PSA and tumor volume decrease. PS-341 (bortezomib, Velcade) is the first proteasome inhibitor with clinical application in cancer therapy that has been used in clinical trials to date. This report reviews the current status of those papers that have tried to analyze the connection between the proteasome pathway and apoptosis. We present our results of proteasome inhibition in individual prostate cancer cell lines. Proteasomal inhibition may offer a new therapeutic access in "molecular targeting" of prostate cancer. PMID- 15526230 TI - [The impact of ALA (5-aminolevulinic acid)-fluorescence detection on the prognosis of superficial bladder cancer]. AB - PURPOSE: Throughout the past years, several studies have shown that fluorescence cystoscopy with ALA (5-aminolevulinic acid) improves the detection rate of superficial bladder tumors by approximately 20 % compared to standard white light cystoscopy. These results suggest a reduced rate of residual/recurrent tumors with the routine use of ALA fluorescence technique prior to bladder tumor resection. The present prospectively randomized study was performed to verify this hypothesis. MATERIALS AND METHODS: A total of 115 bladder tumor patients were randomized for initial resection under white light or ALA fluorescence. After 6 to 8 weeks, a second-look resection was performed in all patients guided by ALA fluorescence. Additional white light cystoscopies were performed after 3, 6 and 12 months. RESULTS: The second-look resection did not find a tumor in 31 of 51 (59 %) patients initially resected under white light guidance compared to 43 of 51 (84 %) patients in the fluorescence group. This difference was statistically significant (p = 0.005). At 12 months, a tumor was not found in 17 of 48 patients from the white light group vs. 25 of 47 patients from the fluorescence group (p = 0.03). Seven patients were lost to follow-up. CONCLUSIONS: By reducing otherwise inevitable re-operations, fluorescence cystoscopy decreases morbidity and lowers treatment costs. PMID- 15526231 TI - [Syndrome of persisting mullerian duct with first manifestation in an adult]. AB - INTRODUCTION: We report on a 22-year-old male patient who presented with an intrapelvic tumor. CASE REPORT: CT and MRI showed a left pelvic abscess, which was drained. After regression of the abscess, we removed the causative tubular structure surgically, revealing a rudimentary genital structure with parts of the Mullerian and Wolffian ducts. CONCLUSIONS: This case report demonstrates an abscess formation as complication of a previously asymptomatic rudimentary genital structure, which was associated with further abnormalities, such as left testicular agenesia, perineoscrotal hypospadias and transverse testicular ectopia. PMID- 15526232 TI - [Breast cancer metastatic to a renal cell carcinoma]. AB - INTRODUCTION: Renal tumors are often diagnosed during routine radiological imaging. A newly diagnosed renal tumor next to an existing cancer is challenging since a primary or a secondary renal neoplasm has to be considered in the differential diagnosis. CASE REPORT: A 64-year-old woman underwent radical mastectomy and axillary lymphadenectomy for cancer of the right breast. After surgery, the patient underwent chemotherapy and radiotherapy because of multiple metastases. Six years later, computed tomography (CT) obtained as follow-up examination revealed a solid mass in the left kidney. Because radiological differentiation between metastatic breast cancer and primary kidney tumor was impossible and fine needle biopsy of renal tumors should be avoided, a nephrectomy was performed. Histologic diagnosis was a metastatic breast cancer within a primary renal cell carcinoma. Whereas the primary tumor was receptor negative, the breast cancer metastasis was estrogen receptor positive. CONCLUSIONS: In a renal mass of unknown nature in patients in good general condition and acceptable life-expectancy, surgical exploration with partial or radical nephrectomy is justified in spite of a synchronous metastatic tumor of different origin. This is the only way to obtain a definitive histologic diagnosis. A primary renal tumor can be treated curatively, preventing secondary complications, such as hematuria. In this case, the changed receptor state of the breast cancer metastasis also offered the patient the possibility of new palliative chemotherapy and hormonal manipulation. PMID- 15526233 TI - [Antireflux plasty using the Lich-Gregoir method]. PMID- 15526236 TI - Current guidelines in the pharmacological management of chronic heart failure. AB - Treatment of heart failure has undergone major changes in the last two decades. European and US guidelines have absorbed these major changes and set the new standards. It is therefore mandatory that treatment of heart failure is based on this knowledge. According to these guidelines, patients with chronic symptomatic heart failure should be treated with a diuretic and sometimes digoxin and vasodilators, to relieve symptoms, in addition to an angiotensin-converting enzyme inhibitor and/or an angiotensin receptor blocker (ARB) and a beta-blocker to improve prognosis. An aldosterone antagonist should be added in patients with more advanced heart failure. Prevention of heart failure has also become a major goal. We summarise here the major issues regarding the pharmacological treatment of chronic heart failure as indicated by the European and US guidelines. PMID- 15526234 TI - Sex-specific genetic architecture of whole blood serotonin levels. AB - Recently, a quantitative-trait locus (QTL) for whole blood serotonin level was identified in a genomewide linkage and association study in a founder population. Because serotonin level is a sexually dimorphic trait, in the present study, we evaluated the sex-specific genetic architecture of whole blood serotonin level in the same population. Here, we use an extended homozygosity-by-descent linkage method that is suitable for large complex pedigrees. Although both males and females have high broad heritability (H2=0.99), females have a higher additive component (h2=0.63 in females; h2=0.27 in males). Furthermore, the serotonin QTL on 17q that was identified previously in this population, integrin beta 3 (ITGB3), and a novel locus on 2q influence serotonin levels only in males, whereas linkage to a region on chromosome 6q is specific to females. Both sexes contribute to linkage signals on 12q and 16p. There were, overall, more associations meeting criteria for suggestive significance in males than in females, including those of ITGB3 and the serotonin transporter gene (5HTT). This analysis is consistent with heritable sexual dimorphism in whole blood serotonin levels resulting from the effects of a combination of sex-specific and sex independent loci. PMID- 15526237 TI - Angiotensin inhibition in heart failure. AB - Survival in patients with heart failure remains very poor, and is worse than that for most common cancers, including bowel cancer in men and breast cancer in women. The renin-angiotensin-aldosterone system (RAAS) is not completely blocked by angiotensin-converting enzyme (ACE) inhibition. Blockade of the RAAS at the AT1-receptor has the theoretical benefit of more effective blockade of the actions of angiotensin II. ACE inhibitors (ACE-Is) prevent the breakdown of bradykinin: this has been blamed for some of the unwanted effects of ACE-Is although bradykinin may have advantageous effects in heart failure. Consequently, ACE-Is and ARBs might be complementary or even additive treatments; recent trials have tested these hypotheses. The Candesartan in Heart failure Assessment of Reduction in Mortality and morbidity (CHARM) programme compared the angiotensin receptor blocker (ARB) candesartan (target dose 32 mg once daily) to placebo in three distinct but complementary populations of patients with symptomatic heart failure. These were: patients with reduced left ventricular ejection fraction (LVEF) who were ACE-I-intolerant (CHARM-Alternative); patients with reduced LVEF who were being treated with ACE-Is (CHARM-Added); and patients with preserved left ventricular systolic function (CHARM-Preserved). There were substantial and statistically significant reductions in the primary composite end point (risk of cardiovascular death or hospital admission for heart failure) in CHARM Alternative. This was also the case in CHARM-Added, supporting and extending the findings of Val-HeFT. In CHARM-Preserved, the effect of candesartan on the primary end point did not reach conventional statistical significance though hospital admission for heart failure was reduced significantly with candesartan. In the CHARM-Overall programme there was a statistically borderline reduction in all-cause mortality with a clear reduction in cardiovascular mortality. All-cause mortality was reduced by 12% in the two CHARM trials in patients with low LVEF. CHARM succeeded in answering a number of questions about the safety and efficacy of ARB use in heart failure. It showed evidence for a clinical benefit of candesartan both additive to and independent of ACE-I use. The benefits in terms of clinical outcomes were seen irrespective of beta-blocker usage. Benefits in patients with preserved LVEF were shown in the proportion of patients hospitalised with worsening heart failure and in overall number of admissions for heart failure. Candesartan had expected effects on blood pressure and renal function, emphasising the need for careful patient monitoring. PMID- 15526238 TI - The epidemiology of heart failure. AB - Heart failure is becoming increasingly common. More than 20 million people worldwide are estimated to have heart failure. Prevalence is rising because the population is ageing: in both men and women, the prevalence of heart failure in those aged 80-89 years is roughly 10 times the prevalence in those aged 50-59 years. Coronary artery disease is now the most common cause of heart failure. Better treatment of myocardial infarction means that more people survive with impaired myocardial function, and some of these will develop heart failure in time. Hypertension is also an important contributing factor. Valvular disease, once a major cause of heart failure, has become less prevalent. The median survival after diagnosis of heart failure was only 1.7 years for men and 3.2 years for women, according to Framingham data for the years 1948 to 1988. After five years, only 25% of men and 38% of women were still alive. Preventive and treatment measures have improved this picture somewhat: deaths from heart failure have decreased by about 12% per decade. However, heart failure continues to carry a grave prognosis. PMID- 15526239 TI - Aldosterone blockade in heart failure. AB - Aldosterone plays a key role in the pathophysiology of heart failure. Angiotensin converting enzyme inhibitors and angiotensin II receptor blockers may not suppress aldosterone production in the long term. This allows aldosterone to exert its effects on myocardial fibrosis and cardiac remodelling, endothelial function, electrolytes and baroreceptor response. The Randomized Aldactone Evaluation Study (RALES) tested spironolactone against placebo in patients with severe heart failure. The study found a 30% reduction in the risk of death among patients treated with spironolactone and a 31% reduction in the risk of death from cardiac causes. Patients in the spironolactone group had significantly lower risks of death from progression of heart failure and sudden cardiac death. The Eplerenone Post-Acute Myocardial Infarction Heart Failure Efficacy and Survival Study (EPHESUS) investigated the effects of eplerenone against placebo in patients with myocardial infarction complicated by left ventricular dysfunction. Compared to placebo, the relative risk of death from any cause was 0.85 in eplerenone-treated patients, and the relative risk of death or hospitalisation for cardiovascular events was 0.87. The reduction in the risk of sudden death from cardiac causes was statistically significant. In conclusion, aldosterone blockade should form part of optimal therapy for patients with heart failure. PMID- 15526240 TI - Reassessing guidelines for heart failure. AB - Significant progress has been made in the last few years in the management of heart failure. In particular, several trials have given significant results. It has become apparent that heart failure may be prevented in some patients by treatment of risk factors such as coronary artery disease. Experience with angiotensin-converting enzyme (ACE) inhibitors has shown that the survival and symptomatic benefits do last in the long term, and confirm that they are the first-line treatment in heart failure. The results of a number of trials using the angiotensin receptor blockers (ARBs) candesartan, valsartan and losartan are presented and discussed. There is also some experience now in the use of candesartan for patients with heart failure and preserved left ventricular systolic function. The COMET trial compared the beta-blockers carvedilol and metoprolol tartrate, and suggests that there may be differences in clinical effect between beta-blockers. The selective aldosterone receptor blocker eplerenone was evaluated in the EPHESUS trial in post-MI patients with signs of heart failure. Based on these clinical trials, heart failure guidelines are now being updated. PMID- 15526241 TI - Conclusions on the management of heart failure. AB - Activity of the renin-angiotensin-aldosterone system (RAAS) is increased in heart failure. Agents which act on the RAAS can be expected to benefit these patients. Recent trials which have furthered our understanding of drugs which are of benefit in heart failure--angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, beta-blockers and aldosterone antagonists--are discussed. It is desirable to use multiple agents for best results in patients with heart failure. New European Society of Cardiology guidelines on the management of heart failure are expected later this year. PMID- 15526242 TI - The role of the renin-angiotensin-aldosterone system in heart failure. AB - Activity of the renin-angiotensin-aldosterone system (RAAS) is increased in patients with heart failure, and its maladaptive mechanisms may lead to adverse effects such as cardiac remodelling and sympathetic activation. Elevated renin activity has been demonstrated in patients with dilated cardiomyopathy. (Third- generation synthetic non-peptide renin inhibitors, with more favourable properties than earlier renin inhibitors, lower ambulatory blood pressure and may have a role to play in other cardiovascular disease.) Chymase, a protease inhibitor stored in mast cells that generates angiotensin II (Ang II) (in addition to angiotensin-converting enzyme [ACE]), has been linked to extracellular matrix remodelling in heart failure. Again, chymase inhibitors have been developed to investigate its functions in vitro and in vivo . Bradykinin is thought to contribute to the cardioprotective effect of ACE inhibition through modification of nitric oxide release, calcium handling and collagen accumulation. Ang II is believed to influence a number of molecular and structural changes in the heart, mostly mediated through the AT1-receptor. The importance of the RAAS in heart failure is shown by the survival benefit conferred by treatment with ACE inhibitors. PMID- 15526243 TI - The VALUE trial: a commentary. AB - The Valsartan Antihypertensive Long-Term Use Evaluation (VALUE) Trial compared coronary heart disease outcome in two anti-hypertensive treatment strategies based on either an angiotensin receptor blocker, valsartan, or a calcium channel blocker (CCB), amlodipine. In both patient groups a diuretic was added, if necessary, in an attempt to achieve blood pressure (BP) goals. Follow-up of over 15,000 patients was maintained for 4.2 years. There were no differences in the primary composite endpoint of cardiac morbidity and mortality (which included interventional procedures, hospitalised heart failure, non-fatal myocardial infarction and fatal coronary heart disease, however myocardial infarction and stroke events occurred less commonly on amlodipine than on valsartan the former achieving statistical significance [p=0.02 and p=0.08 respectively]). There was a non-significant excess of hospitalised heart failure on amlodipine (p=0.012). However, lower BPs early in the trial probably accounted for most of the observed benefits in favour of the CCB. The angiotensin receptor blocker arm was associated with less new onset diabetes. The results of VALUE add further support to the evidence that blood pressure control is the major determinant in outcome in trials of antihypertensive therapy. PMID- 15526244 TI - Arterial stiffness and the renin-angiotensin-aldosterone system. AB - Arterial stiffness has recently been recognised as an independent risk factor for cardiovascular morbidity and mortality in hypertension. Many of the complications seen with angiotensin II (Ang II) excess or hyperaldosteronism--an increased event rate, left ventricular hypertrophy, endothelial dysfunction and target organ damage--are also associated with arterial stiffness. It is possible that reduced arterial compliance may be one mechanism whereby increased activity of the renin-angiotensin-aldosterone system (RAAS) produces adverse vascular effects. Common pathophysiological processes, altered collagen turnover and increased fibrosis may underlie both arterial stiffness and RAAS-associated vascular damage. While it is recognised that patients with hyperaldosteronism have increased arterial stiffness, the role of the RAAS in modulating arterial compliance in essential hypertension and in normotensive subjects is less clear cut. There is, however, more consistent data which show that drugs that interfere with Ang II or aldosterone, namely angiotensin-converting enzyme (ACE) inhibitors, angiotensin receptor blockers (ARBs) and aldosterone antagonists, all reduce arterial stiffness. In many cases, this is to a greater extent than predicted from the extent of reduction in blood pressure (BP), suggesting a role for RAAS in vascular stiffness in hypertensive subjects. There is also evidence that combined ACE inhibitors (ACE-Is) and ARBs may have an additive effect in reducing stiffness. The reduction in cardiovascular mortality in end-stage renal disease patients treated with ACE-Is was preferentially seen in those who had reduced arterial stiffness. These data suggest that, in addition to regulation of vascular biology and BP, the RAAS is an important determinant of arterial stiffness in health and, more particularly, in disease. PMID- 15526245 TI - Statin therapy increases vascular sensitivity to angiotensin II in hypercholesterolaemic patients. AB - Cross-talk between various cardiovascular risk factors has been suggested by a number of studies. This study examines the interaction between hypercholesterolaemia and the renin-angiotensin system in vivo in man. METHODS: We performed a randomised, placebo-controlled, double-blind crossover study on 40 hypercholesterolaemic patients, comparing cholesterol-lowering therapy with a statin for six months versus placebo. Brachial artery function was assessed by bilateral venous occlusion plethysmography using intra-arterial infusions of the endothelial-dependent vasoconstrictors, angiotensin I (Ang I) and angiotensin II (Ang II), to measure vascular angiotensin-converting enzyme (ACE) and Ang II receptor response respectively. The endothelial-independent vasoconstrictor, noradrenaline, was used as a control vasoconstrictor. Results were analysed by multiple analysis of variance and statistical significance was taken as a p value <0.05. RESULTS: Cholesterol-lowering treatment with a statin significantly reduced the mean total cholesterol level to 5.71 mmol/L vs. 7.57 mmol/L on placebo (p<0.0001). Hypercholesterolaemia significantly increased the vasoconstriction response to noradrenaline (placebo versus statin treatment; p=0.046). In hypercholesterolaemia, there was a strong trend towards a reduction in the vasoconstriction response to Ang I (placebo versus statin treatment; p=0.089). In hypercholesterolaemia, the vasoconstriction response to Ang II was significantly reduced (placebo versus statin treatment; p=0.01). CONCLUSIONS: Our in vivo results show that, unlike some other previous work, hypercholesterolaemia is associated with down-regulation of the vasoconstrictor response to Ang II and that statin therapy up-regulates the local vasoconstrictor response to Ang II. The possibility now arises that, in man, statins alter the balance between AT(1) receptors and AT(2)-receptors. PMID- 15526246 TI - Prevention of recurrences in patients with lone atrial fibrillation. The dose dependent effect of angiotensin II receptor blockers. AB - BACKGROUND: Atrial fibrillation (AF) leads to the activation of the renin angiotensin system (RAS), which seems to play an important role in atrial remodelling. It is not known yet whether RAS blockade may prevent recurrences in patients with lone AF. METHODS AND RESULTS: Patients with an episode of persistent AF for >7 days, in the absence of cardiac or extracardiac causes and with normal blood pressure values (lone AF), were recruited. Ninety patients were randomised and scheduled for electrical cardioversion. Three groups of patients were compared: Group I was treated with amiodarone 400 mg daily (30 patients), group II was treated with amiodarone 400 mg daily plus irbesartan 150 mg daily (30 patients) and group III with amiodarone 400 mg daily plus irbesartan 300 mg daily (30 patients). The primary endpoint was the time to a first recurrence of AF. The patients were cardioverted and followed. The Kaplan-Meier analysis of time to first recurrence during the follow-up period showed that patients treated with amiodarone 400 mg plus irbesartan 300 mg had a greater probability of remaining free of AF (77% vs. 52% for amiodarone and 65% for amiodarone+irbesartan 150 mg), hazard ratio for a recurrence in group III: 0.47 (95% CI 0.27-0.82; p=0.001). CONCLUSIONS: The combination of irbesartan plus amiodarone decreased the rate of AF recurrences, with a dose-dependent effect, in lone AF patients. PMID- 15526247 TI - BNP and ANP as diagnostic and predictive markers in heart failure with left ventricular systolic dysfunction. AB - BACKGROUND: The prevalence of chronic heart failure (CHF) with systolic dysfunction is increasing. Plasma natriuretic peptides have been envisaged as diagnostic and predictive markers. AIMS: To investigate the relationship between the levels of B-type natriuretic peptide (BNP) and A-type natriuretic peptide (ANP) and the clinical and functional parameters of CHF in outpatients with CHF at baseline, compared with normal healthy controls; to find out the differences in a randomised controlled trial between patients treated with an angiotensin converting enzyme (ACE) inhibitor, captopril, or an angiotensin receptor blocker (ARB), irbesartan. These differences were assessed throughout the six-month treatment period and at the sixth month. METHODS: Plasma BNP (pmol/L) and ANP (pmol/L) were determined in 68 hypertensive patients with dilated cardiomyopathy, NYHA class III-IV and ejection fraction (EF) < or = 40%, and in 26 normal controls. Statistical analysis for BNP and ANP was done by Students t-test. The patient group was randomly subdivided into two subgroups of 34 patients, each treated with either an ARB, irbesartan, or an ACE inhibitor (ACE-I), captopril. BNP and ANP were measured in both subsamples and correlated with clinical, functional and neurohormonal parameters throughout a follow-up period of six months and at the sixth month. RESULTS: The mean EF in the patient sample was 33.43+/-6.52% and in the controls was 61.96 +/-3.53% (p=0.000). The mean BNP (pmol/L) in patients was 44.78+/-54.36 and in the controls was 7.12+/-8.28 (p=0.000) and the mean ANP (pmol/L) was 30.32+/-25.97 in patients and 11.18+/ 7.92 in controls (p=0.000). A statistically significant difference was found between patients and healthy controls. Significant correlations were found between natriuretic peptides and EF. Between the baseline phase and the sixth month, BNP and ANP decreased significantly in the ARB group. At the sixth month, both BNP and ANP were lower in the ARB group. Evidence of clinical benefit was found with both ARB or ACE-I treatment throughout the six months, with patients moving from classes III and IV to class II NYHA. Improvement of EF was also found, with transition of patients with lower EF (even <30%) to higher values. EF was higher in the ARB group at the sixth month. CONCLUSIONS: BNP and ANP can be useful diagnostic tools in hypertensive CHF patients with moderate-to-severe LV dysfunction. The decrease in BNP and ANP in the ARB group throughout six months, as well as the lower value at the sixth month, suggest a prognostic value of these parameters. PMID- 15526248 TI - Vascular effects of quinapril completely depend on ACE insertion/deletion polymorphism. AB - INTRODUCTION: The angiotensin-converting enzyme (ACE) DD-genotype is associated with increased plasma and myocardial ACE-activity. The influence of the ACE insertion/deletion (I/D) polymorphism on the effects of ACE-inhibition on vascular responses has not been previously described. MATERIALS AND METHODS: In the randomised, double-blind QUinapril On Vascular ACE and Determinants of Ischemia Study (QUO VADIS), 149 patients undergoing coronary bypass surgery were randomised to receive either the ACE inhibitor, quinapril, or placebo. In 82 patients, we obtained ACE-genotype, and measured vascular responses to angiotensin II (Ang II) in left internal mammary arteries. RESULTS: In the placebo group, the mean maximal vasoconstriction to Ang II was significantly lower in patients with the DD-genotype than in those with the ID/II genotype (36.2+/-5.11% [n=13] vs. 55.6+/-4.57% [n=25]; p=0.01). In the quinapril group, the mean maximal vasoconstriction to Ang II was similar between DD- and ID/II genotype (59.6+/-9.19% [n=8] vs. 57.7+/-4.07% [n=35]; p=0.85). CONCLUSIONS: DD genotype patients showed decreased vascular responses to Ang II but treatment with quinapril completely restored the decreased vascular response in DD-genotype patients to the same level as II/ID-genotype patients, while no effect of quinapril was demonstrated in the II/ID-genotype patients. PMID- 15526249 TI - Angiotensin II may mediate apoptosis via AT1-receptors in the rat cardiac conduction system. AB - INTRODUCTION: Apoptosis and angiotensin II (Ang II) have been suggested as possible causes of arrhythmias. In addition, Ang II via Ang II type I (AT(1)-) receptors, has been demonstrated to induce cardiomyocyte apoptosis. The transgenic m(Ren-2)27 (TG) rat carries the additional Ren-2 gene, the expression of which results in an increase in cardiac Ang II, thus potentially affecting the cell growth/death equilibrium. In this study we have investigated the effect of Ang II, via AT(1)-receptors, on mediating apoptosis in a cardiac conduction system (SA node and AV nodes). MATERIALS AND METHODS: Heart sections from male two-day, one-week and two-week TG and Sprague-Dawley (SD) rats were stained with Masson Trichrome to localise the SA and AV nodes. The sections containing SA or AV nodes were processed for quantitation of apoptotic nuclei and AT(1)-receptors. RESULTS: The number of apoptotic nuclei/mm(2) in the SA and AV nodes were found to decrease from two days to two weeks in both the TG and the SD rats, and the number of apoptotic nuclei/mm(2) in the TG groups was significantly higher than that of the SD groups for all ages (p<0.05). The number of AT(1)-receptors/mm(2) in the SA node were found to decrease with increasing age, whereas the number of AT(1)-receptors/mm(2) in the AV node was increased in both TG and SD rats and the number of AT(1)-receptors/mm(2) in the three TG groups was significantly more than that of the three SD groups (p<0.05). DISCUSSION AND CONCLUSION: As a consequence of the additional renin gene in the TG rats, which results in the alteration of the local renin-angiotensin system, the numbers of AT(1) receptors/mm(2) and apoptotic nuclei/mm(2) are increased. The number of apoptotic nuclei/mm(2) and AT(1)-receptors/mm(2) in the SA node decrease with maturation, whereas, the number of AT(1)-receptors in the AV node increase. Thus, there may be a correlation between Ang II and apoptosis in the SA node, which does not appear to be present in the AV node. PMID- 15526250 TI - How different urinary albumin excretion rates can predict progression to nephropathy and the effect of treatment in hypertensive diabetics. AB - HYPOTHESIS: The efficacy of a treatment in a clinical trial depends in part on where the cut-off point is placed for the test result used to select patients for the trial, and this applies to irbesartan in the Irbesartan Microalbuminuria II (IRMA II) trial for preventing nephropathy. PATIENTS AND METHODS: Patients in the IRMA II trial were stratified into different pre-treatment albumin excretion rate (AER) ranges to compare the proportion of patients starting in these different ranges (i) that progressed to develop nephropathy within 24 months and (ii) whose AER was over 40 microg/minute at three months. RESULTS: The proportion of patients with pre-treatment AER values between 20 and 40 microg/minute progressing to develop nephropathy was 1.25% in the placebo group and 0.78% in the irbesartan group, while for pre-treatment AER values between 41 and 200 microg/minute, 24.4% and 11.2% develop nephropathy respectively in the placebo and irbesartan groups. In patients with a pre-treatment AER of 20 to 30 microg/minute, 32.5% and 13.6% respectively in the placebo and irbesartan groups had a value exceeding 40 microg/minute at three months. CONCLUSIONS: The data demonstrate that irbesartan is effective in reducing the onset of nephropathy within two years when the pre-treatment AER is above 40 microg/minute, but if the AER is below this level it progresses unusually to nephropathy within two years. Irbesartan also slows progression of AER to over 40 microg/minute for patients with pre-treatment AER values at or above 20 microg/minute and these patients should be treated. PMID- 15526251 TI - Renin-angiotensin system blockade prevents the increase in plasma transforming growth factor beta 1, and reduces proteinuria and kidney hypertrophy in the streptozotocin-diabetic rat. AB - INTRODUCTION: Combination therapy with angiotensin-converting enzyme (ACE) inhibitors and angiotensin receptor blockers (ARBs) is used to improve renal outcome achieved by monotherapy in diabetic patients. In addition, interference with the renin-angiotensin system (RAS) reduced expression and excretion of transforming growth factor beta 1 (TGF-beta 1) in diabetic nephropathy. The aim of this study was to investigate the effects of interrupting the RAS by ACE inhibitor (ACE-I) or ARB monotherapy or by combination therapy on proteinuria, kidney hypertrophy and plasma TGF-beta 1 in diabetic rats. MATERIALS AND METHODS: Forty-one male Wistar rats were allocated to five groups: 1 = control rats, 2 = diabetic rats (streptozotocin [STZ] 55 mg/kg), 3 = diabetic rats as above receiving enalapril (20 mg/kg/day), 4 = diabetic rats receiving losartan (80 mg/kg/day), 5 = diabetic rats receiving both losartan and enalapril. The study lasted 60 days. RESULTS: Urinary protein excretion, kidney weight, serum ACE activity and plasma TGF-beta1 increased significantly in untreated diabetic rats compared with controls. Administration of losartan, enalapril, or both for 60 days prevented these changes. Furthermore, combined therapy for 30 days normalised urinary protein excretion, while monotherapy did not. Losartan inhibited serum ACE activity both in vivo and in vitro. Plasma TGF-beta 1 levels were positively correlated with blood glucose levels (r=0.4059) and with urinary protein excretion (r=0.3558). CONCLUSIONS: Combination therapy with losartan and enalapril was more effective than monotherapy with either drug in achieving an early antiproteinuric response. Long-term treatment with losartan was as effective as the combined treatment, possibly due to a dual inhibitory effect on the RAS. The antiproteinuric effect may be related, in part, to reduced TGF-beta 1. PMID- 15526252 TI - Psychosocial impairment and recurrence of major depression. AB - Major depressive disorder is often marked by multiple recurrences. Unfortunately, it is difficult to predict specifically which patients will suffer a recurrence. This study examined whether psychosocial impairment is a risk factor for recurrence. A total of 290 subjects with unipolar major depressive disorder according to Research Diagnostic Criteria (RDC) were prospectively followed for up to 15 years as part of the Collaborative Depression Study (CDS), a multicenter longitudinal observational study of the mood disorders. Follow-up data on course of illness and psychosocial functioning were collected with the Longitudinal Interval Follow-up Evaluation (LIFE). The association of psychosocial impairment with recurrence of major depression was examined with mixed-effects logistic regression. The mean (SD) score for psychosocial functioning during recovery from an episode of major depression was 9.0 (2.7), with a possible range of 4 (no impairment) to 20 (severe impairment). For euthymic subjects who recovered from an episode of major depression, elevated psychosocial impairment was significantly associated with subsequent recurrence of major depression, with an odds ratio of 1.12 (95% confidence interval [CI], 1.06 to 1.19). The odds ratio of 1.12 indicates that for each 1-point increase in the functional impairment score, the risk of recurrence increases by about 12%. In patients who have recovered from an episode of major depression, the presence of psychosocial impairment may help identify who is at increased risk of recurrence. PMID- 15526253 TI - Longitudinal course of bipolar I disorder. AB - The course of bipolar I disorder is characterized by frequently fluctuating levels of manic and depressive symptoms. In the current study, we sought to characterize the month-by- month course of this disorder in 61 patients who were originally enrolled in a clinical trial and were followed for a mean of 23.7 months (SD = 6.1). All patients in the trial received medication management; some received family psychosocial interventions as well. On a monthly basis, we assessed symptom severity using the Modified Hamilton Rating Scale for Depression (MHRSD) and the Bech-Rafaelson Mania Scale (BRMS). Each month, we categorized each participant as fully symptomatic, partially symptomatic, or asymptomatic in terms of both depressed and manic symptoms. We found that the median percent time fully symptomatic was 8%, the median percent time partially symptomatic was 22%, and the median percent time asymptomatic was 59%. Using DSM-IV-TR criteria for defining an acute mood episode, we found that the median length of episode was 1 month, and participants experienced, on average, one episode every 8 months. Estimates concerning percent time fully symptomatic and asymptomatic converge with those reported in other datasets. PMID- 15526254 TI - Factor structure of recalled DSM-IV hypomanic symptoms of bipolar II disorder. AB - The DSM-IV-TR definition of hypomania in bipolar II disorder (BP-II) has yet to show its validity. The aim of the current study was to find the factor structure of hypomania by using DSM-IV-TR symptoms, and to assess the DSM-IV-TR definition of hypomania. One hundred ninety-seven consecutive BP-II remitted outpatients were interviewed by the Structured Clinical Interview for DSM-IV (SCID-CV) as modified by Benazzi and Akiskal (2003) and by Benazzi (2003), in a private practice, assessing the symptoms that were more common during past hypomanic episodes. The factor structure of hypomania was studied by principal component factor analysis. Almost all patients reported overactivity (increased goal directed activity) during hypomania, and less commonly elevated mood. Overactivity plus three or more symptoms identified 89.3% of DSM-IV-TR BP-II. Factor analysis found three factors: factor 1, including racing thoughts ("mental activation"); factor 2, including elevated mood ("high mood"); and factor 3, including overactivity ("behavioral activation"). Elevated mood was correlated only with two of the nine DSM-IV-TR hypomanic symptoms. The three-domains structure of hypomania by Kraepelin (i.e., increased mood, thought, and activity) was found in the DSM-IV-TR definition of hypomania, partly supporting its list of symptoms. However, DSM-IV-TR priority given to mood change for the diagnosis of hypomania was not supported. An upgrading of overactivity to at least a priority level similar to mood change was supported by (1) its high frequency, (2) its utility to diagnose BP-II, and (3) by factor analysis showing that elevated mood (the "prototypical" symptom of hypomania in DSM-IV-TR) correlated with few symptoms, and that three factors (of which only one included elevated mood) were present. PMID- 15526255 TI - Attempted suicide and self-injury in patients diagnosed with eating disorders. AB - Eating disorders (ED) patients are at high risk for developing suicidal behavior. The aim of the present study was to investigate factors associated with suicidal behavior in ED patients. One hundred fifty patients at an outpatient ED clinic were included in the study. Data were gathered by retrospective chart analysis. We found that 48 patients (32%) had a history of parasuicide (i.e., suicide attempts, self-injury, or both). A significantly greater percentage of parasuicidal patients than nonparasuicidal patients had EDs with bingeing/pursing symptomatology, used more than one type of purging method, and had a lifetime history of a drug use disorder, impulse control problems, and bipolar disorder, as well as a more extensive outpatient and inpatient treatment history. The findings of this study support an increased tendency toward impulsivity among parasuicidal ED outpatients. PMID- 15526256 TI - Attentional disturbances in patients with unipolar psychotic depression: a selective and sustained attention study. AB - Psychotic depression is a clinical subtype of major depressive disorder in the recent editions of the psychiatric diagnostic systems ICD-10 (1992) and DSM-IV (1994). Recent evidence suggests that psychotic depressed patients are more impaired on neuropsychologic tests measuring attention as compared to nonpsychotic depressed patients. However, information on this issue between psychotic and nonpsychotic depression is limited. It has become clear that attention is not a single concept; thus we studied both selective and sustained attention using the theoretic model of automatic and controlled information processing. Thirty-two patients with major depressive disorder, 16 psychotics and 16 nonpsychotics, were investigated and compared with 20 patients with schizophrenic disorder and 20 healthy volunteers who comprised the control groups, using Ruff's 2 and 7 selective attention tests. Compared to the healthy controls, both depressed groups were impaired; however, the psychotic depressed group was more severely impaired on both measures. Attentional performance speed and accuracy scores, on both effortless and effortful conditions, were significantly lower in the psychotic depressed group than in the nonpsychotic depressed group. No significant differences were found on attentional performance between the psychotic depressed patients and those with schizophrenic disorder. Attention deficits are thus more prominent in psychotic than in nonpsychotic depression. Furthermore, taking attention as a criterion, psychotic depression, although of mood congruent subtype, lies closer to schizophrenia than to nonpsychotic depression. PMID- 15526257 TI - Anxiety disorders in schizophrenia. AB - Data regarding the co-occurrence of anxiety symptoms or syndromes in schizophrenia is scant. Hierarchical assumptions embedded in diagnostic systems and methodologic difficulties hamper the development of studies on accessory symptomatology outside of the core positive-negative-disorganized symptoms. Recent studies have repeatedly challenged these assumptions by presenting data on comorbid disorders in schizophrenia. We review the current knowledge about anxiety comorbidity in schizophrenia, and its relative prevalence, relevance, and treatment. A computerized search of the literature published from 1966 to July 2003 was conducted on Medline using the word "schizophrenia" and the words from the names of each anxiety disorder listed in DSM-IV, one at a time. Only studies including exclusively the diagnosis of schizophrenia were included. Only 15 studies were dedicated to the subject of anxiety comorbidity prevalence in schizophrenia. The most studied comorbidities were obsessive-compulsive disorder (OCD) and panic disorder, and most reports suggested them to be highly prevalent in schizophrenia. Posttraumatic stress disorder (PTSD) was the least studied (N = 2). Except for two pharmacologic studies in OCD, there were no double-blind randomized controlled trials regarding the treatment of anxiety in schizophrenia. Most case reports and open label trials tried conventional treatment for anxiety disorders with some degree of success. The literature indicates that anxiety comorbidities are prevalent in schizophrenia and conventional treatment for anxiety can help alleviate the symptoms in those patients. However, the body of data provided by research so far is still far from allowing evidence-based conclusions. Large studies with operationalized criteria, as well as comprehensive assessments of treatment response and outcome are needed to clarify the relationship between anxiety disorders and schizophrenia. PMID- 15526258 TI - Expressed emotion and social functioning in chronic schizophrenia. AB - The aim of this study was to reveal the relationships between family expressed emotion (EE), family evaluations of social functioning, and the psychopathologic symptoms of patients with schizophrenia. We examined whether EE influenced the social functioning of patients with schizophrenia. Forty-four subjects with schizophrenia and 82 of their relatives participated in this study. The Five Minute Speech Sample (FMSS) was conducted to evaluate EE, and subjects were divided into high-EE and low-EE groups. The Positive and Negative Symptom Scale (PANSS) was used to assess symptom severity. Social functioning was compared between the two groups using the Social Functioning Scale (SFS). No differences in symptom severity or social adjustment, as evaluated by a global assessment of functioning, were observed between the two groups. However, the high-EE relatives tended to evaluate the social functioning of the schizophrenia patient in their family as being rather low and showed a strong dissatisfaction with the patient's social withdrawal and level of independence (competence). Furthermore, low-EE relatives in high-EE families showed the same tendencies. The family members who were evaluated as low-EE relatives in a high-EE family were dissatisfied with the patient's social withdrawal, level of independence (competence), and also their interpersonal functioning. In the chronic stable phase of schizophrenia, the attention of the family members may be more directed towards changes in social functioning; thus, the EE may reflect a family's attitude towards improvements in the patient's social functioning. PMID- 15526259 TI - Complicated grief in Bosnian refugees: associations with posttraumatic stress disorder and depression. AB - Complicated grief is likely to be common among refugee populations exposed to war trauma. However, there have been few studies investigating the traumatic antecedents and correlates of complicated grief in refugees, and the relationship of that symptom pattern with other common disorders such as posttraumatic stress disorder (PTSD) and depression. We studied Bosnian refugees recruited from a community center in Sydney, Australia, with the sample being supplemented by a snowball method (N = 126; response rate, 86%). Measures included a trauma inventory, the Clinician Administered PTSD Scale (CAPS), the depression module of the Structured Clinical Interview (SCID), and the Core Bereavement Items (CBI). A dimension of traumatic loss derived from the trauma inventory was a specific predictor of complicated grief, with exposure to human rights violations being associated with images of the traumatic events surrounding the lost person. There was no link between PTSD and grief other than for a low-order association with the PTSD intrusion dimension. In contrast, depression was strongly associated with grief and its subscales. Only the subgroup with comorbid grief and depression reported higher levels of traumatic loss. The results suggest that complicated grief in refugees can become persistent and associated with depression. While PTSD and grief share common symptoms of intrusion, the two symptom domains are sufficiently distinct to warrant independent assessment of grief in refugee populations. PMID- 15526260 TI - Italian version of the defense style questionnaire. AB - The Defense Style Questionnaire (DSQ) assesses defensive behavior by empirically evaluating conscious derivatives of defense and coping mechanisms in everyday life. It was developed on the assumption that defenses can be ordered along a maturity-immaturity continuum and tend to group into clusters, or defensive styles. The original factor analytical study, by Bond et al. (1983) identified four styles, called maladaptive, image-distorting, self-sacrifice, and adaptive styles. Successive studies only partially confirmed this factor structure. We present the factor structure and the main psychometric features of the Italian version of the questionnaire. The DSQ was translated into Italian by the back translation method and administered to a sample of 294 men (mean age, 33.33 years) and 333 women (mean age, 32.38 years). An exploratory factor analysis identified three factors largely corresponding to Bond's maladaptive, image distorting, and adaptive defensive styles and to analogous factors identified by other authors. Accordingly, three defense style scales were constructed, containing respectively 37, 17, and 12 items. These scales showed intercorrelations compatible with the hierarchical model of defensive functioning at the base of the questionnaire, acceptable, though ameliorable, test-retest reliabilities (r's = .79, .63, and .81, respectively) and, with the exception of the Adaptive Style scale, sufficient internal consistencies (alphas: .85, .72, .57). However, only the Maladaptive Style scale, probably due to its greater length, showed values of reliability and internal consistency high enough to warrant clinical use in its present form. Further investigation is required to find new items that may improve the reliability of the Image-Distorting and the Adaptive Style scales. PMID- 15526262 TI - Impact of aminopolycarboxylates on aquatic organisms and eutrophication: overview of available data. AB - Aminopolycarboxylic acids, which include ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA), diethylenetriaminepentaacetic acid (DTPA), 1,3 propylenediaminetetraacetic acid (1,3-PDTA), beta-alaninediacetic acid (beta ADA), and methylglycinediacetic acid (MGDA), constitute a class of complexing agents that occur in a wide range of domestic products and that are used intensively as metal sequestrants in several industrial applications. Because they are highly polar and partially nondegradable, aminopolycarboxylates are released into the aquatic environment in significant quantities, mainly via wastewater. The historical and current use of aminopolycarboxylates and their ubiquitous presence in surface waters prompted many studies about their possibly detrimental impact on aquatic organisms. This review summarizes the available data and information on the eutrophication potential and toxicity of aminopolycarboxylates to a multitude of aquatic organisms including vertebrates, invertebrates, algae, bacteria, and protozoa. This article also addresses how the ecotoxic effects of aminopolycarboxylates are dependent on their speciation, that is, on their presence in a free or a metal-complexed form. PMID- 15526263 TI - Ultrastructural changes in various organs of the fish Puntius gonionotus fed cadmium-enriched cyanobacteria. AB - The accumulation and toxicity of cadmium in Puntius gonionotus fish that consumed the cyanobacterium Spirulina platensis contaminated with cadmium were studied. Fish were fed cadmium-contaminated cells for 4 weeks, after which cadmium accumulation in various organs was determined. The highest cadmium content was found in the kidney (56.0 microg Cd/g wet weight). Cadmium was not detected in the gill during the entire 4 weeks of cadmium feeding. Histopathological alteration of cells was observed in the gill, kidney, and liver. The results showed that dietary cadmium caused hypertrophy and edema of gill filaments. Coagulative necrosis and karyolysis of the nucleus were observed in the kidney. Vacuoles and hyaline droplets had accumulated in the epithelial cells of the proximal tubule. In the liver vacuolation of the cytoplasm, infiltration of macrophages, and focal necrosis were found. The ultrastructural changes that occurred in the cells of different organs were similar. These included a proliferation of vacuoles and lysosomes, formation of myelin bodies, degranulation, vesiculation, and dilation of rough endoplasmic reticulum, as well as swelling of mitochondria with loss of cristae. PMID- 15526264 TI - Histological evidence of intersex in feral sharptooth catfish (Clarias gariepinus) from an estrogen-polluted water source in Gauteng, South Africa. AB - This is the first histological evidence of intersex in a fish species inhabiting a South African water source. One hundred catfish (Clarias gariepinus) were collected randomly from the Marais Dam (MD) and the Rietvlei Dam (RVD) in the Rietvlei Nature Reserve (RNR), South Africa. These dams drain water from a stream that receives effluent from industrial sites, agricultural activities, informal settlements, and municipal treatment plants. Each fish was evaluated macroscopically and had blood drawn, and its gonads were macroscopically and histologically examined to verify intersex potentially related to endocrine disruption. Gonadal histology of several fish showed primary oocytes scattered in testicular tissue, indicative of intersex. The results showed intersexuality in 20% of fish from both the dams. The GSI value for intersex fish was closer to male GSI values, suggesting that the sampled intersex fish were more likely to have occurred from the feminization of male catfish. Target chemical analyses showed that the water, sediment, and serum samples tested positive for p nonylphenol (p-NP). The p-NP level in water and sediment at MD was 6360 and 4.0 microg/kg, respectively, whereas in sediment at RVD it was 113 microg/kg. Commonly found in the effluent from sewage treatment plants, p-NP in water and sediment indicates estrogenic water pollution, which might affect wildlife and humans dependent on these sources. PMID- 15526265 TI - Detection and quantification of genotoxicity in wastewater-treated Tetrahymena thermophila using the comet assay. AB - In the present study, the comet, or single-cell, gel electrophoresis assay was adapted for use with the ubiquitous unicellular protozoan Tetrahymena thermophila, and the method was evaluated for its ability to detect DNA damage induced by known genotoxins and wastewater samples. The original comet assay protocol was substantially modified (e.g., lower concentrations of detergents were used in the lysis buffer; electrophoresis time was reduced). Using the modified method, T. thermophila were subjected to short exposures of phenol, hydrogen peroxide, and formaldehyde, leading to concentration-dependent increases in DNA damage. The genotoxic potential of influent and effluent water samples from a local municipal wastewater treatment plant was evaluated. The results indicated that the influent wastewater was genotoxic and that the genotoxicity in the effluent water was substantially reduced. We assume employing T. thermophila in the use of the comet assay may become a cost-effective and reliable tool for genotoxicity screening and monitoring of wastewater and similar systems. PMID- 15526266 TI - Physiological and biochemical analyses of microcystin-RR toxicity to the cyanobacterium Synechococcus elongatus. AB - Freshwater Microcystis may form dense blooms in eutrophic lakes. It is known to produce a family of related cyclic hepatopeptides (microcystins, MC) that constitute a threat to aquatic ecosystems. Most toxicological studies of microcystins have focused on aquatic animals and plants, with few examining the possible effects of microcystins on phytoplankton. In this study we chose the unicellular Synechococcus elongatus (one of the most studied and geographically most widely distributed cyanobacteria in the picoplankton) as the test material and investigated the biological parameters: growth, pigment (chlorophyll-a, phycocyanin), photosynthetic activity, nitrate reductase activity, and protein and carbohydrate content. The results revealed that microcystin-RR concentrations above 100 microg x L(-1) significantly inhibited the growth of Synechococcus elongatus. In addition, a change in color of the toxin-treated algae (chlorosis) was observed in the experiments. Furthermore, MC-RR markedly inhibited the synthesis of the pigments chlorophyll-a and phycocyanin. A drastic reduction in photochemical efficiency of PSII (F(v)/F(m)) was found after a 96-h incubation. Changes in protein and carbohydrate concentrations and in nitrate reductase activity also were observed during the exposure period. This study aimed to evaluate the mechanisms of microcystin toxicity on a cyanobacterium, according to the physiological and biochemical responses of Synechococcus elongatus to different doses of microcystin-RR. The ecological role of microcystins as an allelopathic substance also is discussed in the article. PMID- 15526267 TI - Light-emitting diodes for the illumination of algae in ecotoxicity testing. AB - Ecotoxic effects of chemicals, preparations, and environmental samples are routinely quantified in standardized algae growth inhibition tests using microalgae like Desmodesmus subspicatus. The intention of this investigation was to prove the suitability of light-emitting diodes (LEDs) as an alternative to tubular discharge lamps in algae growth inhibition tests. This was approached by demonstrating that growth of algae was similar with both illuminating devices. Growth factors of growth controls were determined using LEDs. Growth curves of different light intensities of LEDs were compared with those of tubular discharge lamps. By testing certain chemicals with both illumination methods comparable results for E(r)C50 values were obtained. It could be shown that small-scale construction of incubators using LEDs is possible and could be advantageous, especially for the illumination of algae in 96-well microplate growth inhibition assays. PMID- 15526268 TI - Cytometric analysis of adverse action of diphenyl ditelluride on rat thymocytes: cell shrinkage as a cytotoxic parameter. AB - Despite the growing use of organotellurium compounds in the chemical and biomedical fields, there has been no great concern about their toxicity until now. To test the possibility that diphenyl ditelluride (DPDT) and tellurium chloride (TeCl2), organic and inorganic tellurium compounds, may exert adverse action on mammals, their effects on rat thymocytes were examined under in vitro conditions using a flow cytometer with fluorescent probes. Incubation of thymocytes with DPDT at 300 nM or more for 24 h significantly increased the populations of shrunken cells and of cells with hypodiploidal DNA. Z-VAD-FMK, a paninhibitor of caspases, greatly suppressed the DPDT-induced increase in the hypodiploidal cell population, suggesting the involvement of caspase activation in DPDT toxicity. Hence, it is possible that DPDT would increase the population of thymocytes undergoing apoptosis if the blood concentration in mammals reached at least 300 nM or more. TeCl2 was much less potent than DPDT in increasing the population of hypodiploidal cells. PMID- 15526269 TI - Evaluation of the mutagenic potential of urban air pollution in Sao Paulo, Southeastern Brazil, using the Tradescantia stamen-hair assay. AB - We used a short-term bioassay--the Tradescantia stamen-hair assay (TSH)--to evaluate the toxicity of ambient particles with an aerodynamic diameter less than 10 microm sampled in the metropolitan region of Sao Paulo, Brazil. Two exposure locations were selected: downtown Sao Paulo and Caucaia do Alto (mean PM10 levels of 64 and 14 microg/m3, respectively). The experiment was conducted July 11 August 15, 2002, and toxicity was assessed with the Tradescantia stamen-hair assay (TSH) employing clone KU-20 of Tradescantia. Four experimental groups were defined: inflorescences collected from plants cultivated in Caucaia, inflorescences collected from plants cultivated in Sao Paulo (to establish the baseline level of mutations in stamen hairs at both sites), inflorescences collected from plants cultivated in Caucaia and brought to Sao Paulo and maintained in chambers that received ambient air, and inflorescences collected from plants cultivated in Caucaia and brought to Sao Paulo and maintained in chambers that received air passed through a particle filter. The frequency of mutations observed in Caucaia was significantly lower than that in the remaining groups. Flower cuttings brought from Caucaia and receiving ambient air of Sao Paulo showed a rate of mutations similar to that of plants cultivated in Sao Paulo. Filtering particles from the air reduced the rate of mutation but not sufficiently to reach the level of that in Caucaia. The frequency of mutations observed in Sao Paulo was significantly associated with PM10 levels on the fifth day before the opening of the flowers (r = 0.47, p = 0.025). Our results indicate that urban particles play a significant role in the development of pollution dependent mutations. PMID- 15526270 TI - Inflammatory activity in river-water samples. AB - Contamination of the urban aquatic environment with chemical and biological substances could have a long-term impact on human health because these substances threaten the integrity of the urban ecosystem and the availability of high quality water for recreation and consumption. In light of this, the aim of the present study was to assess the potential immunological effects of water sampled at various sites along the River Saale near the city of Halle (in the state of Sachsen-Anhalt, Germany). For the control, Ficoll-separated peripheral blood mononuclear cells (PBMC) of healthy donors were cultured for 24 h in either filter-sterilized river water or drinking-water samples. Cell vitality was assessed using the MTT bioassay. Cytokines in culture supernatants were measured by ELISA. Endotoxin concentrations in the water samples were assessed by the limulus amoebocyte lysate (LAL) test. River water and drinking water showed comparably weak cytotoxic effects on PBMC. Drinking water did not exert any effect on cytokine secretion. In contrast, all river-water samples triggered secretion of proinflammatory cytokines, as shown for TNF-alpha, IL-1beta, and IL 6. Free endotoxin was detected in all river-water samples. However, the highest inflammatory activity regarding induction of all three cytokines, as well as the highest endotoxin content as determined by LAL, was found in a water sample taken immediately downstream of a wastewater treatment plant. Inhibition studies using the monoclonal anti-CD14 antibody biG14, which is known to suppress binding of lipopolysaccharide (LPS) to CD14 via binding CD14 itself, revealed that free endotoxin was indeed the major inducer of proinflammatory cytokines in the river water samples. Taken together, the results suggest that the microorganism-derived endotoxin is a widely distributed contaminant in the urban aquatic environment that should be considered in routine monitoring and in assessing ecosystem and human health. PMID- 15526271 TI - Effects of in vitro exposure to low levels of organotin and carbamate pesticides on human natural killer cell cytotoxic function. AB - Human natural killer (NK) lymphocytes play a central role in immune system defense against viral infection and against the formation of primary tumors. Organotin (OT) pesticides have been used in industrial and agricultural applications, and OT contamination has been reported in water, sediment, and fish. Carbamate pesticides are currently used in agricultural chemicals. Two specific carbamates used in agriculture are ziram and maneb; ziram also is used as an additive in rubber products including latex gloves. In previous studies we demonstrated that at concentrations in the 150-200 nM range, the OTs tributyltin (TBT) and triphenyltin (TPT) were capable of disrupting the function of human NK cells after incubations to as short as 24 h. Previously, we also examined the effects of ziram and maneb at higher concentrations on the cytotoxic function of human NK cells. The current study examined the effects of exposure of up to 6 days to lower concentrations of each of these compounds on the cytotoxic function of NK cells. The OTs were studied at concentrations ranging from 200 to 10 nM; ziram was studied at concentrations of 2.5 microM-125 nM and maneb at concentrations of 10-1 microM. These conditions were studied both in highly purified NK cells and in a mixture of lymphocytes containing both T and NK cells. As little as 25 nM TBT decreased the function of purified NK cells after 24 and 48 h, whereas 10 nM TBT was effective after 6 days. The lowest level of TPT that was effective at 24 h was 50 nM whereas the results after 48 h and 6 days were similar to those seen with TBT. The presence of T lymphocytes diminished the effects of both TBT and TPT on NK cytotoxic function. A concentration of ziram as low as 125 nM produced significant loss of cytotoxic function in highly purified NK cells (65% decrease in function after 6 days). The toxicity of each of the compounds studied increased very significantly with length of exposure. PMID- 15526272 TI - Comparative study of microcystin-LR-induced behavioral changes of two fish species, Danio rerio and Leucaspius delineatus. AB - The spontaneous locomotor behavior separated into day/night activity phases of two fish species Danio rerio and Leucaspius delineatus was recorded and quantified continuously under sublethal long-term exposure to microcystin-LR in tanks. Microcystin-LR was applied in concentrations of 0.5, 5, 15, and 50 microg L(-1). By using an automated video-monitoring and object-tracing system, the average motility (swimming velocity) and the average number of turns were assessed. Clear dose-dependent effects of microcystin-LR on the behavior of both test fish were measured. During the daytime, the motility of Danio rerio as well as Leucaspius delineatus increased significantly by exposure to the lowest concentrations, whereas higher concentrations led to significantly decreased motility. Influenced by microcystin-LR, the swimming time of Leucaspius delineatus reversed, going from a prominently diurnal activity to a nocturnal one; Danio rerio remained active during the daytime. Most of the relative changes in the behavioral patterns of Danio rerio and Leucaspius delineatus suggest these fish have comparable susceptibility to microcystin-LR and may indicate some adverse consequences for fish populations, for example, in connection with reproduction and predator-prey interactions. PMID- 15526273 TI - Identification of progressive cervical epithelial cell abnormalities using DNA image cytometry. AB - BACKGROUND: The objectives of the current study were to compare the capabilities of conventional cervical cytology and of DNA image cytometry (DNA-ICM) in the prediction of progressive or regressive behavior in atypical squamous cells (ASC), low-grade squamous intraepithelial lesions (LSIL), and atypical glandular cells (AGC). METHODS: One hundred ninety-six women with Papanicolaou (Pap) smears that yielded diagnoses of ASC, LSIL, or AGC were included in a prospective cohort study. Slides were classified according to the Bethesda system. DNA-ICM was performed according to the consensus reports of the European Society of Analytical Cellular Pathology. RESULTS: Reference standard verification was available in 108 patients. The rate of DNA aneuploidy in Pap smears increased significantly from cervical intraepithelial neoplasia 1 (CIN1) (54%) and CIN2 (64.3%) to CIN3 or greater (CIN3+) (83.3%) in subsequent biopsies (P < 0.05). Using ASC, LSIL, and AGC as input cytologic diagnoses and >/= CIN2 as the output histologic diagnosis, the positive predictive values (PPVs) for conventional cytology and DNA-ICM were 35.2% and 65.9%, respectively (P < 0.001). The negative predictive value (NPV) of DNA-ICM was 85.0%. When >/= CIN3 was used as the output histologic diagnosis, conventional cytology had a PPV of 22.2%. The PPV and NPV of DNA-ICM were 43.9% and 93.3%, respectively. CONCLUSIONS: The results of the current study confirmed the prognostic validity of DNA image cytometry for differentiation between progressive and regressive lesions in patients with ASC, LSIL, and AGC diagnoses. PMID- 15526274 TI - Beta-catenin and BMP-2 synergize to promote osteoblast differentiation and new bone formation. AB - Mutations of critical components of the Wnt pathway profoundly affect skeletal development and maintenance, probably via modulation of beta-catenin signaling. We tested the hypothesis that beta-catenin is involved in mesenchymal lineage allocation to osteogenic cells using a beta-catenin mutant with constitutive transcriptional activity (DeltaN151). Although this stable beta-catenin had no effects by itself on osteogenic differentiation of multipotent embryonic cell lines, it synergized with bone morphogenetic protein-2 (BMP-2) resulting in dramatic stimulation of alkaline phosphatase activity, osteocalcin gene expression, and matrix mineralization. Likewise, DeltaN151 and BMP-2 synergistically stimulated new bone formation after subperiosteal injection in mouse calvaria in vivo. Conversely, DeltaN151 prevented adipogenic differentiation from pre-adipocytic or uncommitted mesenchymal cells in vitro. Intriguingly, the synergism with BMP-2 on gene transcription occurred without altering expression of Cbfa1/Runx2, suggesting actions independent or downstream of this osteoblast-specific transcription factor. Thus, beta-catenin directs osteogenic lineage allocation by enhancing mesenchymal cell responsiveness to osteogenic factors, such as BMP-2, in part via Tcf/Lef dependent mechanisms. In vivo, this synergism leads to increased new bone formation. PMID- 15526277 TI - Doxazosin inhibits human vascular endothelial cell adhesion, migration, and invasion. AB - The quinazoline-derived alpha1-adrenoceptor antagonists, doxazosin and terazosin have been recently shown to induce an anoikis effect in human prostate cancer cells and to suppress prostate tumor vascularity in clinical specimens [Keledjian and Kyprianou, 2003]. This study sought to examine the ability of doxazosin to affect the growth of human vascular endothelial cells and to modulate vascular endothelial growth factor (VEGF)-mediated angiogenesis. Human umbilical vein endothelial cells (HUVECs) were used as an in vitro model to determine the effect of doxazosin on cell growth, apoptosis, adhesion, migration, and angiogenic response of endothelial cells. The effect of doxazosin on cell viability and apoptosis induction of human endothelial cells, was evaluated on the basis of trypan blue and Hoechst 33342 staining, respectively. Doxazosin antagonized the VEGF-mediated angiogenic response of HUVEC cells, by abrogating cell adhesion to fibronectin and collagen-coated surfaces and inhibiting cell migration, via a potential downregulation of VEGF expression. Furthermore there was a significant suppression of in vitro angiogenesis by doxazosin on the basis of VEGF-mediated endothelial tube formation (P < 0.01). Fibroblast growth factor-2 (FGF-2) significantly enhanced HUVEC cell tube formation (P < 0.01) and this effect was suppressed by doxazosin. These findings provide new insight into the ability of doxazosin to suppress the growth and angiogenic response of human endothelial cells by interfering with VEGF and FGF-2 action. This evidence may have potential therapeutic significance in using this quinazoline-based compound as an antiangiogenic agent for the treatment of advanced prostate cancer. PMID- 15526278 TI - Sustained diacylglycerol accumulation resulting from prolonged G protein-coupled receptor agonist-induced phosphoinositide breakdown in hepatocytes. AB - Studies in various cells have led to the idea that agonist-stimulated diacylglycerol (DAG) generation results from an early, transient phospholipase C (PLC)-catalyzed phosphoinositide breakdown, while a more sustained elevation of DAG originates from phosphatidylcholine (PC). We have examined this issue further, using cultured rat hepatocytes, and report here that various G protein coupled receptor (GPCR) agonists, including vasopressin (VP), angiotensin II (Ang.II), prostaglandin F2alpha, and norepinephrine (NE), may give rise to a prolonged phosphoinositide hydrolysis. Preincubation of hepatocytes with 1 butanol to prevent conversion of phosphatidic acid (PA) did not affect the agonist-induced DAG accumulation, suggesting that phospholipase D-mediated breakdown of PC was not involved. In contrast, the GPCR agonists induced phosphoinositide turnover, assessed by accumulation of inositol phosphates, that was sustained for up to 18 h, even under conditions where PLC was partially desensitized. Pretreatment of hepatocytes with wortmannin, to inhibit synthesis of phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate (PIP2), prevented agonist-induced inositol phosphate and DAG accumulation. Upon VP stimulation the level of PIP) declined, but only transiently, while increases in inositol 1,4,5-trisphosphate (InsP3) and DAG mass were sustained, suggesting that efficient resynthesis of PIP2 allowed sustained PLC activity. This was confirmed when cells were pretreated with wortmannin to prevent resynthesis of PIP2. Furthermore, metabolism of InsP3 was rapid, compared to that of DAG, with a more than 20-fold difference in half-life. Thus, rapid metabolism of InsP3 and efficient resynthesis of PIP2 may account for the larger amount of DAG generated and the more sustained time course, compared to InsP3. The results suggest that DAG accumulation that is sustained for many hours in response to VP, Ang.II, NE, and prostaglandin F2alpha in hepatocytes is mainly due to phosphoinositide breakdown. PMID- 15526279 TI - Endothelial cell myosin light chain kinase (MLCK) regulates TNFalpha-induced NFkappaB activity. AB - Tumor necrosis factor (TNFalpha-) generates both apoptotic and survival signals with endothelial cell (EC) survival dependent on nuclear factor kappa-B (NFkappaB) activation, a regulator of anti-apoptotic genes. We previously demonstrated that increased EC contractility, rearrangement of the actin cytoskeleton, and increased myosin light chain (MLC) phosphorylation occurs as a consequence of TNFalpha-induced activation of EC MLC kinase (EC MLCK) and is required for bovine lung EC apoptosis. As the association between MLCK and pro survival signals such as NFkappaB activation is unknown, we studied the role of MLCK in the regulation of NFkappaB-dependent transactivation in bovine pulmonary artery EC. Both TNFalpha-induced increase in NFkappaB dependent transactivation measured by NFkappaB luciferase reporter assay (approximately fivefold) and nuclear translocation of NFkappaB were significantly inhibited by MLCK-selective inhibitors, KT5926 (60% inhibition of luciferase activity) and ML7 (50% decrease). Furthermore, our data revealed that inhibition of MLCK attenuated the TNFalpha-induced IkappaB phosphorylation, translocation of p65, NFkappaB-DNA binding, and NFkappaB transcriptional activity. Molecular approaches to either reduce EC MLCK expression (AdV EC MLCK antisense construct) or to reduce kinase activity (kinase-dead EC MLCK ATPdel mutant) produced similar attenuation of the TNFalpha-induced NFkappaB response. In contrast, a constitutively active MLCK mutant (EC MLCK1745) enhanced TNFalpha-induced luciferase activity. Together, these novel observations indicate that TNFalpha-induced cytoskeletal rearrangement driven by MLCK activity is necessary for TNFalpha-dependent NFkappaB activation and amplification of pro-survival signals. PMID- 15526280 TI - Signaling function of PSGL-1 in neutrophil: tyrosine-phosphorylation-dependent and c-Abl-involved alteration in the F-actin-based cytoskeleton. AB - P-selectin glycoprotein ligand-1 (PSGL-1) is the best-characterized selectin ligand that has been demonstrated to mediate leukocytes rolling on endothelium and leukocytes recruitment into inflamed tissue in vivo. In addition to its direct role in leukocyte capturing, PSGL-1 also functions as a signal-transducing receptor. The present work showed that after cross-linking of PSGL-1 with KPL1, an anti-PSGL-1 monoclonal antibody, PSGL-1 linked to the cytoskeleton and became a detergent-insoluble component in activated neutrophils. The antibody cross linking led to the polymerization and redistribution of F-actin-based cytoskeleton, and this alteration of cytoskeleton was spatiotemporally related to the polarization of PSGL-1. PSGL-1's polarization was cytoskeleton-dependent because it was eliminated by cytochalasin B. Furthermore, the polymerization and redistribution of F-actin filaments were tyrosine-phosphorylation-dependent since the alteration of F-actin-based cytoskeleton was severely blocked by genistein, a universal tyrosine kinase inhibitor. STI571, a small molecule inhibitor for cytoplasmic tyrosine kinase c-Abl, also inhibited the alteration of F-actin-based cytoskeleton, and c-Abl was redistributed to where F-actin concentrated in the activated neutrophils. The results suggested that cross-linking of PSGL-1 induces the phosphorylation-dependent and c-Abl-involved alteration of F-actin-based cytoskeleton in neutrophils. PMID- 15526281 TI - Bone morphogenetic protein-2 induces expression of murine zinc finger transcription factor ZNF450. AB - The bone morphogenetic protein-2 (BMP-2) is a potent secreted factor that promotes osteoblast differentiation during development. Exposure to BMP-2 is sufficient to cause a lasting change in cell fate presumably by activating specific target genes. To identify genes downstream of BMP-2 we treated the murine pluripotent embryonic cell line, C3H10T1/2 that can be induced to form an osteoblastic phenotype, with 100 ng/ml BMP-2 for 24 h. Using suppression subtractive hybridisation we found the novel zinc finger transcription factor, ZNF450 was upregulated. The single-copy ZNF450 gene spans 15.6 kb on chromosome 10B1 and consists of seven exons, the first of which is untranslated. The open reading frame encodes a 710 reside protein. Analysis of the protein sequence reveals a highly conserved amino-terminal BTB/POZ dimerisation domain, an AT-hook motif, and eight C2H2 zinc fingers. Library screening identified a second mRNA isoform encoding a short protein isoform with one zinc finger. Using reverse transcriptase-real time PCR to measure mRNA expression we found that ZNF450, Runx2/Cbfa-1, and Sp7/osterix were induced by BMP-2 after 4 h in C2C12 myoblast cells. Treatment of C2C12 cells with BMP-2 causes a shift from a myoblastic to osteoblastic phenotype. ZNF450 was upregulated three to fivefold after 24 h in C3H10T1/2 cells and required 100 ng/ml BMP-2. Expression of the 3 kb major transcript was highest in liver, testis, and kidney. However, ZNF450 mRNA was found also in a wide range of adult tissues. The consistent induction of ZNF450 by BMP-2 after 4 h in three murine pluripotent cell lines suggests that ZNF450 may play a role in the BMP-2 signalling pathway. PMID- 15526282 TI - Suppression of cyclic GMP-specific phosphodiesterase 5 promotes apoptosis and inhibits growth in HT29 cells. AB - Phosphodiesterase 5 (PDE5) is a major isoform of cGMP phosphodiesterase in a variety of human tumor cell lines and plays a key role in regulating intracellular cGMP concentrations ([cGMP]i). Here, we demonstrate that suppression of PDE5 gene expression by antisense pZeoSV2/ASP5 plasmid transfection results in a sustained increase in [cGMP]i, growth inhibition, and apoptosis in human colon tumor HT29 cells. With stable transfection, antisense transcripts exhibited a specific suppression in PDE5 activity, mRNA levels, and a 93 kDa hPDE5A1 protein. In cloned antisense cells, prolongation of the cell growth doubling times correlate positively with suppressed PDE5 activity and increased [cGMP]i. The growth inhibition in PDE5 antisense clones is due to an increased apoptotic rate and delayed cell-cycle progression. These results corroborate previous findings with the PDE5 inhibitor exisulind and its derivatives showing that sustained [cGMP]i induces apoptosis and growth inhibition in tumor cells. Furthermore, an inducible mitotic inhibitor p21WAF1/CIP1 has been found to account for the delay of cell-cycle progression in PDE5 antisense clones at G2/M phase. A proteolytic cleavage of p21WAF1/CIP1 in the antisense clones is also increased at the later stage of serum stimulation. The protein kinase G (PKG) inhibitor, KT5823, can prevent the cleavage of p21(WAF1/CIP). These data substantiate a pivotal role for PDE5 as a modulator of apoptosis and cell-cycle progression for human carcinoma via a mechanism involving the activation of [cGMP]i/PKG signaling pathways. PMID- 15526283 TI - Differentiation of human colon adenocarcinoma cells alters the expression and intracellular localization of annexins A1, A2, and A5. AB - Butyrate induces differentiation and alters cell proliferation in intestinal epithelial cells by modulation of the expression of several genes. Annexins are a superfamily of ubiquitous proteins characterized by their calcium-dependent ability to bind to biological membranes; their involvement in several physiological processes, such as membrane trafficking, calcium signaling, cell motility, proliferation, and differentiation has been proposed. Thus, we have analyzed changes in annexin A1 (AnxA1), annexin A2 (AnxA2), and annexin A5 (AnxA5) levels and localization in human colon adenocarcinoma cells differentiated by butyrate treatment or by culture in glucose-free inosine containing medium. The acquired differentiated phenotype increased dipeptidyl peptidase-IV (DPP-IV) expression and alkaline phosphatase (ALP) activity, two well known brush border markers. Butyrate induces cell differentiation and growth arrest in BCS-TC2, BCS-TC2.2, HT-29, and Caco-2 cells, increasing the levels of AnxA1 and AnxA5, whereas AnxA2 decreases except in Caco-2 cells. Inosine differentiated cells present increased amounts of the three studied annexins, as occurs in spontaneously differentiated Caco-2 cells. AnxA2 down-regulation is not due to proteasome activation and seems to be related to the butyrate-induced cell proliferation arrest; AnxA1 and AnxA5 expression is growth-state independent. AnxA1 and AnxA5 are mainly found in the cytoplasm while AnxA2 is localized underneath the plasma membrane in cell-to-cell contacts. Butyrate induces changes in subcellular localization towards a vesicle-associated pattern. Human colon adenocarcinoma cell differentiation is associated with an up-regulation of AnxA1, AnxA2, and AnxA5 and with a subcellular relocation of these proteins. No correlation between annexin levels and tumorigenicity was found. Up-regulation of AnxA1 could contribute to the reported anti-inflammatory effects of butyrate in colon inflammatory diseases. PMID- 15526284 TI - Up-regulation of the association between heat shock protein 90 and endothelial nitric oxide synthase prevents high glucose-induced apoptosis in human endothelial cells. AB - Hyperglycemia is the hallmark of diabetes mellitus. Poor glycemic control is correlated with increased cardiovascular morbidity and mortality. High glucose can trigger endothelial cell apoptosis by de-activation of endothelial nitric oxide synthase (eNOS). eNOS was recently demonstrated to be extensively regulated by Akt and heat shock protein 90 (HSP90). Yet, little is known about the molecular mechanisms that regulate eNOS activity during high glucose exposure. The present study was designed to determine the involvement of protein interactions between eNOS and HSP90 in high glucose-induced endothelial cell apoptosis. The protein interaction of eNOS/HSP90 and eNOS/Akt were studied in cultured human umbilical vein endothelial cells (HUVECs) exposed to either control-level (5.5 mM) or high-level (33 mM) glucose for different durations (2, 4, 6, and 24 h). The results showed that the protein interactions between eNOS and HSP90 and between eNOS and Akt and the phosphorylation of eNOS were up regulated by high glucose exposure for 2-4 h. With longer exposures, these effects decreased gradually. During early hours of exposure, the protein interactions of eNOS/HSP90 and eNOS/Akt and the phosphorylation of eNOS were all inhibited by geldanamycin, an HSP90 inhibitor. High glucose-induced endothelial cell apoptosis was also enhanced by geldanamycin and was reversed by NO donors. LY294002, a phosphatidylinositol 3 (PI3) kinase inhibitor, inhibited the association of eNOS/Akt and the phosphorylation of eNOS but had no effect on the interaction between eNOS and HSP90 during early hours of exposure. From our results we propose that, in HUVECs, during early phase of high glucose exposure, apoptosis can be prevented by enhancement of eNOS activity through augmentation of the protein interaction between eNOS and HSP90 and recruitment of the activated Akt. With longer exposure, dysregulation of eNOS activity would result in apoptosis. The present study provides a molecular basis for the effects of eNOS in the prevention of endothelial cells apoptosis during early phase of high glucose exposure. These observations may contribute to the understanding of the pathogenesis of vascular complications in diabetes mellitus. PMID- 15526285 TI - Transcriptional activation of the testis-specific histone H1t gene by RFX2 may require both proximal promoter X-box elements. AB - The rat testis-specific linker histone H1t gene is transcribed in pachytene primary spermatocytes during spermatogenesis. Our previous work using transgenic mice demonstrated that spermatocyte-specific transcription of the H1t gene is dependent upon a proximal promoter element designated the TE element. TE is composed of two adjacent and inverted imperfect repeat sequences designated TE1 and TE2 and both of these palindromic elements are similar in sequence to the X box, a DNA consensus sequence that binds regulatory factor X (RFX). RFX2 is the major enriched protein derived from rat testis nuclear extracts when using the TE1 element as an affinity chromatography probe. Co-expression of RFX2 together with an H1t promoted reporter vector in transient expression assays activates the H1t promoter in the GC-2spd germinal cell line, and mutation of either X-box significantly represses activity. However, RFX2 partially reactivates the promoter when either of the X-box elements is independently mutated. In order to totally block reactivation by RFX2, it is necessary to mutate both X-boxes simultaneously. Therefore, RFX2 appears to be able to bind to either X-box independently to partially activate the promoter of the testis-specific histone H1t gene, but simultaneous binding of RFX2 to both X-box elements may be required for maximal promoter activation. PMID- 15526286 TI - Cellular organization and appearance of differentiated structures in developing stages of the parasitic platyhelminth Echinococcus granulosus. AB - Echinococcus granulosus is the causative agent of hydatidosis, a major zoonoses that affects humans and herbivorous domestic animals. The disease is caused by the pressure exerted on viscera by hydatid cysts that are formed upon ingestion of E. granulosus eggs excreted by canine. Protoscoleces, larval forms infective to canine, develop asynchronously and clonally from the germinal layer (GL) of hydatid cysts. In this report, we describe the cellular organization and the appearance of differentiated structures both in nascent buds and developed protoscoleces attached to the GL. Early protoscolex morphogenesis is a highly complex and dynamic process starting from the constitution of a foramen in the early bud, around which nuclei are distributed mainly at the lateral and apical regions. Similarly, distribution of nuclei in mature protoscoleces is not homogenous but underlies three cellular territories: the suckers, the rostellar pad, and the body, that surrounds the foramen. Several nuclei are associated to calcareous corpuscles (Cc), differentiated structures that are absent in the earlier bud stages. The number of nuclei is similar from the grown, elongated bud stage to the mature protoscolex attached to the GL, strongly suggesting that there is no significant cellular proliferation during final protoscolex development. The amount of DNA per nucleus is in the same range to the one described for most other platyhelminthes. Our results point to a sequential series of events involving cell proliferation, spatial cell organization, and differentiation, starting in early buds at the GL of fertile hydatid cysts leading to mature protoscoleces infective to canine. PMID- 15526287 TI - Compensating bends in a 16-base-pair DNA oligomer containing a T(3)A(3) segment: A NMR study of global DNA curvature. AB - In-phase ligated DNA containing T(n)A(n) segments fail to exhibit the retarded polyacrylamide gel electrophoresis (PAGE) migration observed for in-phase ligated A(n)T(n) segments, a behavior thought to be correlated with macroscopic DNA curvature. The lack of macroscopic curvature in ligated T(n)A(n) segments is thought to be due to cancellation of bending in regions flanking the TpA steps. To address this issue, solution-state NMR, including residual dipolar coupling (RDC) restraints, was used to determine a high-resolution structure of [d(CGAGGTTTAAACCTCG)2], a DNA oligomer containing a T3A3 tract. The overall magnitude and direction of bending, including the regions flanking the central TpA step, was measured using a radius of curvature, Rc, analysis. The Rc for the overall molecule indicated a small magnitude of global bending (Rc = 138 +/- 23 nm) towards the major groove, whereas the Rc for the two halves (72 +/- 33 nm and 69 +/- 14 nm) indicated greater localized bending into the minor groove. The direction of bending in the regions flanking the TpA step is in partial opposition (109 degrees), contributing to cancellation of bending. The cancellation of bending did not correlate with a pattern of roll values at the TpA step, or at the 5' and 3' junctions, of the T3A3 segment, suggesting a simple junction/roll model is insufficient to predict cancellation of DNA bending in all T(n)A(n) junction sequence contexts. Importantly, Rc analysis of structures refined without RDC restraints lacked the precision and accuracy needed to reliably measure bending. PMID- 15526289 TI - Further characterization of the genetic defect of the Bent tail mouse, a mouse model for human neural tube defects. AB - BACKGROUND: Neural tube defects (NTDs) are congenital malformations arising mostly from incomplete neural tube closure during early embryogenesis. Most NTDs in humans have a complex etiology, with involvement of both genetic and environmental factors. More than 100 mouse models for human neural tube defects exist; Bent tail is one of them. The mouse mutant is caused by a submicroscopic deletion on Xq that completely encompasses the Zic3 gene. METHODS: We searched the ENSEMBL database for other genes/transcribed sequences in the Bent tail deletion in addition to Zic3, which we confirmed by PCR analysis. RESULTS: In our study, we show that the Bent tail deletion is at least 300 kb in size, encompassing a processed pseudogene and a number of expressed sequence tags in addition to Zic3. Although more research is needed to clarify the identity and function of the deleted transcripts, most of them are expressed during embryonic development and might therefore contribute to the phenotype of the Bent tail mouse. CONCLUSIONS: This study presents the first evidence for the fact that the Bent tail allele is not merely a Zic3 knockout allele, as has been previously suggested. PMID- 15526290 TI - Molecular genetic analysis of human homologs of Caenorhabditis elegans mab-21 like 1 gene in patients with neural tube defects. AB - BACKGROUND: Neural tube defects (NTDs) are complex embryological malformations, affecting 1 in 1,000 live births. Antisense studies have implicated murine Mab21 genes as having an important role in neural tube development. We investigated whether MAB21L1/L2 genes could be involved in the aetiology of NTDs. METHODS: Denaturing HPLC (DHPLC) analysis of MAB21 genes was performed in 116 NTD cases. A case-control approach was used to test if the two single nucleotide polymorphisms (SNPs) of the MAB21L1 gene might be associated with increased NTD risk. RESULTS: No pathological variants of MAB21L1/L2 genes were identified by DHPLC analysis. Case-control studies demonstrated that the two SNPs (CAG triplets in 5'UTR; A-->C in 3'UTR) in the MAB21L1 gene are unlikely to be directly responsible for myelomeningocele. CONCLUSIONS: We suggest that MAB21 genes are unlikely to have substantial impact on NTDs. These preliminary findings will need to be investigated in larger samples before firm conclusions can be made. PMID- 15526288 TI - Genistein stimulates the osteoblastic differentiation via NO/cGMP in bone marrow culture. AB - The soybean phytoestrogen, genistein (Gen), has anabolic effects on bone through mechanisms that remain to be elucidated. We examined the role of nitric oxide (NO) and its downstream effector guanylyl cyclase (GC) in mediating the effects of Gen on the proliferation and osteoblastic maturation of primary mouse bone marrow-derived mesenchymal stem cells (BMSCs). Gen (10(-8) approximately 10(-6) M) resulted in a dose-dependent increase in cell proliferation as measured by increased [3H]thymidine incorporation, and stimulated osteoblastic maturation as assessed by culture duration-dependent increments in alkaline phosphatase (ALP) activity, calcium deposition into extracellular matrix and Runx2/Cbfa1 gene expression in BMSCs cultures. Gen also resulted in a dose-dependent increase in NO synthase (NOS) activity, NO formation, and cGMP production in BMSCs cultures. The effects of Gen were mimicked by 17beta-estradiol (E2, 10(-8) M). Concurrent treatment with the estrogen receptor (ER) antagonist ICI182,780 (10(-7) M) or the NOS inhibitor L-NAME (3 x 10(-3) M) diminished the Gen (10(-6) M)-mediated increase in NOS activity, NO production, and cGMP content. In contrast, a soluble GC inhibitor 1H-[1,2,4]oxadiazolo [4,3,-a]quinoxalin-1-one (ODQ, 10(-6) M) selectively blocked the Gen (10(-6) M)-mediated increase in cGMP content but not in NO production and NOS activity. Moreover, inhibition of ER, NOS activity or cGMP blocked Gen-induced proliferation and osteoblastic differentiation of BMSCs and Runx2/Cbfa1 gene expression in culture. Gen has estrogen-like activity and stimulates the proliferation and osteoblastic differentiation of mouse BMSCs at least in part through NO/cGMP pathway. PMID- 15526291 TI - De novo interstitial direct duplication of Xq21.1q25 associated with skewed X inactivation pattern. AB - Genotype-phenotype correlation in women with an abnormal phenotype associated with a duplication of the long arm of the X chromosome remains unclear. We report on prenatal diagnosis and follow-up of a girl with an Xq duplication and dysmorphic features. The abnormal phenotype included growth retardation, hypotonia, and nystagmus. In order to improve the resolution of the cytogenetic analysis, we used both conventional and array-based comparative genomic hybridization to perform a global molecular cytogenetic analysis of the genome. These molecular cytogenetic analyses showed a direct duplication Xq21.1 --> q25 without other chromosomal abnormalities. This duplication was originating from the paternal X chromosome. Moreover, a skewed X-inactivation pattern was observed leading to a partial functional disomy of the chromosomal region Xq21.1q25. This report and review of the literature suggest that functional disomy for chromosome X could explain the abnormal phenotype. In prenatal diagnosis, this can have implication for patient management and genetic counseling. PMID- 15526292 TI - Reduced birth defects caused by maternal immune stimulation in methylnitrosourea exposed mice: association with placental improvement. AB - BACKGROUND: Methylnitrosourea (MNU) is a potent carcinogen and teratogen that is associated with central nervous system, craniofacial, skeletal, ocular, and appendicular birth defects following transplacental exposure at critical time points during development, and preliminary studies have suggested that nonspecific maternal immunostimulation may offer protection against development of these birth defects. METHODS: Our study examined morphologic alterations in fetal limb and digital development and placental integrity following maternal exposure to MNU on GD 9 in CD-1 mice, and characterized the improvement in placental integrity and abrogation of fetal defects following maternal immune stimulation with interferon-gamma (IFN-gamma) on GD 7. RESULTS: Fetal limbs were significantly shortened (p < 0.0001) and incidence of limb and digital defects (syndactyly, polydactyly, oligodactyly, clubbing, and webbing) was dramatically increased following mid-gestational maternal MNU exposure. Maternal immune stimulation with IFN-gamma on GD 7 lessened incidence of fetal limb shortening and maldevelopment on GD 12 and 14. Further, disruption of placental spongiotrophoblast integrity, increased cell death in placental trophoblasts with increased intercellular spaces in the spongiotrophoblast layer and minimal inflammation, and increased loss of fetal labyrinthine endothelial cells from MNU exposed dams suggested that MNU-induced placental breakdown may contribute to fetal limb and digital maldevelopment. MNU + IFN-gamma was associated with diminished cell death within all layers of the placenta, especially in the labyrinthine layer. CONCLUSIONS: These data verify improved distal limb development in MNU-exposed mice as a result of maternal IFN-gamma administration, and suggest a link between placental integrity and proper fetal development. PMID- 15526293 TI - Iron-containing nodules of cirrhosis. AB - In the absence of genetic hemochromatosis and systemic hemosiderosis, patients with cirrhosis can accumulate focal iron within regenerative or dysplastic hepatic nodules, commonly referred to as 'siderotic nodules'. Siderotic dysplastic nodules are premalignant lesions while siderotic regenerative nodules are a marker for severe viral or alcoholic cirrhosis. The relationship of hepatic iron deposition to hepatic cirrhosis and neoplasia has not been fully clarified. This article will review the current literature regarding selective iron accumulation in siderotic nodules in chronic liver disease, followed by a discussion of current MR imaging techniques for detection and characterization of these nodules. PMID- 15526294 TI - Localization of MRX82: a new nonsyndromic X-linked mental retardation locus to Xq24-q25 in a Basque family. AB - Clinical and molecular studies are reported on a Basque family (MRX82) with nonsyndromic X-linked mental retardation (XLMR) in five affected males. A total of 38 microsatellite markers were typed. The XLMR locus has been linked to DXS8067, DXS1001, DXS425, DXS7877, and DXS1183 with a maximum LOD score of 2.4. The haplotype studies and multipoint linkage analysis suggest a localization of the MRX82 locus to an interval of 7.6 Mb defined by markers DXS6805 and DXS7346, in Xq24 and Xq25, respectively. No gene contained in this interval has been so far associated with nonsyndromic mental retardation, except for GRIA3, disrupted by a balanced translocation in a female patient with bipolar affective disorder and mental retardation. However, the search for mutations of this gene did not showed a pathogenic mutation in the present family. Given that there are other eight MRX families overlapping this interval, none of them with known mutation, we conclude that at least one new gene responsible for nonsyndromic mental retardation is located in this region. PMID- 15526295 TI - Phase II study of temozolomide without radiotherapy in newly diagnosed glioblastoma multiforme in an elderly population. PMID- 15526297 TI - Anchor profiles of HLA-specific peptides: analysis by a novel affinity scoring method and experimental validation. AB - The study of intermolecular interactions is a fundamental research subject in biology. Here we report on the development of a quantitative structure-based affinity scoring method for peptide-protein complexes, named PepScope. The method operates on the basis of a highly specific force field function (CHARMM) that is applied to all-atom structural representations of peptide-receptor complexes. Peptide side-chain contributions to total affinity are scored after detailed rotameric sampling followed by controlled energy refinement. A de novo approach to estimate dehydration energies was developed, based on the simulation of individual amino acids in a solvent box filled with explicit water molecules. Transferability of the method was demonstrated by its application to the hydrophobic HLA-A2 and -A24 receptors, the polar HLA-A1, and the sterically ruled HLA-B7 receptor. A combined theoretical and experimental study on 39 anchor substitutions in FxSKQYMTx/HLA-A2 and -A24 complexes indicated a prediction accuracy of about two thirds of a log-unit in Kd. Analysis of free energy contributions identified a great role of desolvation and conformational strain effects in establishing a given specificity profile. Interestingly, the method rightly predicted that most anchor profiles are less specific than so far assumed. This suggests that many potential T-cell epitopes could be missed with current prediction methods. The results presented in this work may therefore significantly affect T-cell epitope discovery programs applied in the field of peptide vaccine development. PMID- 15526298 TI - ATP-induced structural change of dephosphocoenzyme A kinase from Thermus thermophilus HB8. AB - Dephosphocoenzyme A kinase (DCK) catalyzes phosphorylation in the final step of coenzyme A (CoA) biosynthesis. In this phosphorylation process, domain movements play a very important role. To reveal the structural changes induced by ligand binding, we determined the crystal structure of DCK from Thermus thermophilus HB8 by the multiwavelength anomalous dispersion method at 2.8 A. The crystal structure includes three independent protein molecules in the asymmetric unit: One is a liganded form and the others are unliganded. The topology shows a canonical nucleotide-binding protein possessing the P-loop motif. A structure homology search by DALI revealed the similarity of the DCKs from T. thermophilus HB8, Haemophilus influenzae, and Escherichia coli. Structural comparisons between the liganded and unliganded forms of DCK from T. thermophilus HB8 indicated domain movements induced by adenosine triphosphate (ATP) binding. For the domain movements, proline residues confer flexibility at the domain linkages. In particular, Pro91 plays an important role in moving the CoA domain. PMID- 15526299 TI - Early events in protein aggregation: molecular flexibility and hydrophobicity/charge interaction in amyloid peptides as studied by molecular dynamics simulations. AB - In a previous article (Zbilut et al., Biophys J 2003;85:3544-3557), we demonstrated how an aggregation versus folding choice could be approached considering hydrophobicity distribution and charge. In this work, our aim is highlighting the mutual interaction of charge and hydrophobicity distribution in the aggregation process. Use was made of two different peptides, both derived from a transmembrane protein (amyloid precursor protein; APP), namely, Abeta(1 28) and Abeta(1-40). Abeta(1-28) has a much lower aggregation propensity than Abeta(1-40). The results obtained by means of molecular dynamics simulations show that, when submitted to the most "aggregation-prone" environment, corresponding to the isoelectric point and consequently to zero net charge, both peptides acquire their maximum flexibility, but Abeta(1-40) has a definitely higher conformational mobility than Abeta(1-28). The absence of a hydrophobic "tail," which is the most mobile part of the molecule in Abeta(1-40), is the element lacking in Abeta(1-28) for obtaining a "fully aggregating" phenotype. Our results suggest that conformational flexibility, determined by both hydrophobicity and charge effect, is the main mechanistic determinant of aggregation propensity. PMID- 15526300 TI - Porcine beta-lactoglobulin chemical unfolding: identification of a non-native alpha-helical intermediate. AB - The chemical unfolding behavior of porcine beta-lactoglobulin (PLG) has been followed at pH 2 and 6 in the presence of guanidinium hydrochloride. The PLG unfolding transition, monitored by tryptophan fluorescence, far and near UV circular dichroism and 1D-NMR, can be described by a three-state transition suggesting the presence of at least one intermediate state that appears to display an excess of non-native alpha-helical structures. The thermodynamic parameters, as determined through a global analysis fitting procedure, give estimates of the free energy differences of the transitions connecting the native, the intermediate and the unfolded state: DeltaG(NI) (0) = 2.8 +/- 0.7 kcal mol(-1) (pH 2) and 4.2 +/- 0.5 kcal mol(-1) (pH 6) and DeltaG(NU) (0) = 7.2 +/- 0.6 kcal mol(-1) (pH 2) and 6.9 +/- 0.6 kcal mol(-1) (pH 6). CD unfolding data of the bovine species (BLG) have been collected here under the same experimental conditions of PLG to allow a careful comparison of the two beta lactoglobulins. Intermediates with different characteristics have been identified for BLG and PLG, and their nature has been discussed on a structural analysis basis. The thermodynamic data reported here for PLG and BLG and the comparative analysis with data reported for equine beta lactoglobulin, show that homologous beta-barrel proteins, belonging to the same family and displaying high sequence identity (52-64%) populate unfolding intermediates to different extents, even though a common tendency to the formation of non-native alpha-helical intermediates, can be envisaged. The present results provide a prerequisite foundation of knowledge for the design and interpretation of future folding kinetic studies. PMID- 15526301 TI - Adaptation of the ACE for a Malayalam speaking population in southern India. AB - OBJECTIVE: To adapt the Addenbrooke's Cognitive Examination (ACE) as a dementia screening tool in a community in south India. To establish that items in the adapted version are equivalent to that in the original. METHODS: The ACE was adapted into the local language, Malayalam (m-ACE), following cultural/linguistic modifications. To establish equivalence, qualitative comparisons were made (on the distribution of scores, percentage scoring at ceiling, and relative difficulty across items) between a UK sample receiving the ACE (n = 50; mean age = 67.9 +/- 7.4; education >/= 9, mean = 10.9 +/- 2.5) and a community-based educationally-stratified Indian sample receiving the m-ACE: 'India >/= 9' (n = 50; mean age = 67.8 +/- 5.2; education >/= 9, mean = 13.9 +/- 2.7) and 'India /= 9 groups. Language, Naming, Attention and Orientation are relatively easy (>/= 80% at ceiling) and Recall and Verbal fluency are relatively difficult ( 23 and no word remembered on the aMMSE delayed-recall task (SMI group); and (2) aMMSE > 23 and at least one word remembered on the delayed-recall task (control group). In a three-year follow-up wave, conversion rate to dementia was calculated and logistic regression was performed. RESULTS: Of a total of 2507 subjects who completed the two evaluations, 280 qualified for SMI at entry. In the SMI group, 25 subjects (8.9%) developed dementia vs 26 subjects (1.2%) in the control group. Taking the two groups together, and once demographic and medical variables had been controlled, a low delayed-recall score increased dementia conversion rate (OR 0.47, 95% CI 0.34-0.64). Alzheimer's disease was the main cause of dementia (79.8%). CONCLUSIONS: Memory impairment is a risk factor for future dementia in the neurologically-healthy elderly. This can be observed in a subgroup of subjects with SMI defined on the aMMSE delayed-recall subscore. Some other measurements should be added to the SMI construct to improve its predictive validity. PMID- 15526310 TI - A pilot study of behavioural and psychological signs and symptoms of dementia in patients of Indian sub-continent origin admitted to a dementia day hospital in the United Kingdom. AB - BACKGROUND: There is a paucity of cross-cultural studies of behavioural and psychological symptoms of dementia (BPSD). METHOD: BPSD were examined in a consecutive series of Indian sub-continent origin and white indigenous elders admitted to a dementia day hospital using the BEHAVE-AD. The correlates of individual BPSD in each of the two ethnic groups and the differences between the two ethnic groups were examined. RESULTS: There were no differences between the two groups on most of the demographic and clinical variables examined, except that Indian sub-continent elders had a greater number of children. There were no differences between the two groups on the MMSE scores, BEHAVE-AD total scores and BEHAVE-AD subscale scores (with one exception). Indian sub- continent origin patients had lower scores on the anxiety and phobias subscale. Within the Indian sub-continent origin group, Alzheimer's disease (AD) was associated with activity disturbance and vascular dementia with affective disturbance. Within the indigenous group, aggressivity was associated with males and prescription of neuroleptics, and affective disturbance with prescription of antidepressants. CONCLUSION: There is a need to develop and evaluate translated versions of instruments that measure BPSD. After development of these instruments there is a need for cross-cultural population-based epidemiological studies of BPSD. PMID- 15526311 TI - Sugar-poly(para-phenylene ethynylene) conjugates as sensory materials: efficient quenching by Hg2+ and Pb2+ ions. AB - Three polar poly(para-phenylene ethynylene)s (PPE) were synthesized by utilizing the Heck-Sonogashira protocol. Two of the PPEs carry beta-glucopyranose substituents. Depending upon the linker used between the glycol units and the backbone, the fluorescence of these PPEs can be quenched by Hg2+ and Pb2+ to a varying degree. Monomeric model compounds that are substituted with only one glucose unit are not efficiently quenched. The presence of many glucose substituents in one PPE assembly led to a large increase in the binding constant to Hg2+ and quenching of the fluorescence was amplified. PMID- 15526313 TI - A remarkable ligand orientational effect in osmium-atom-induced blue phosphorescence. AB - A new series of Os(II)-based carbonyl complexes cis(CO),trans(Npy,Npy),cis(Ntz,Ntz)-[Os(CO)2(bptz)2] (1), cis(CO),cis(Npy,Npy),trans(Ntz,Ntz)-[Os(bptz)2(CO)2] (2), and cis(CO),trans(Npy,Npy),cis(Ntz,Ntz)-[Os(CO)2(fptz)2] (3), where bptz and fptz denote 3-tert-butyl-5-(2-pyridyl)- and 3-trifluoromethyl-5-(2-pyridyl)-1,2,4 triazolate, respectively, have been designed and synthesized in an effort to achieve high efficiency, room-temperature blue phosphorescence. Although 1 and 2 are geometric isomers, remarkably different excited-state relaxation pathways were observed. Complex 1 exhibits strong phosphorescence in CH3CN (Phi(p) approximately 0.47) and as a single crystal at room temperature, whereas complex 2 is nearly nonemissive under similar conditions. The associated relaxation dynamics have been comprehensively investigated by spectroscopic and relaxation dynamics as well as by theoretical approaches. Our results lead us to the conclusion that for complex 2, the "loose bolt" effect of metal-ligand bonding interactions plays a crucial role in the fast radiationless deactivation of this type of geometrical isomer. Fine adjustment can also be achieved by functionalizing the ligands so that the electron-withdrawing nature of the CF3 group in 3 stabilizes the HOMO of the triazolate moiety, thus moving the emission further into the pure "blue" region; this results in highly efficient phosphorescence and renders 3 particularly attractive for application in blue OLED devices. PMID- 15526312 TI - No association between subjective memory complaints and apolipoprotein E genotype in cognitively intact elderly. AB - OBJECTIVE: This cross-sectional study examined the relationship between subjective memory complaints and the apolipoprotein epsilon 4 allele (epsilon4), a genetic risk factor for Alzheimer's disease (AD), among cognitively normal subjects identified from a community memory screening. DESIGN: The sample comprised 232 consecutive white non-Hispanic older adults who presented to a free community-based memory-screening program at a University affiliated memory disorders center. Participants were classified as cognitively normal based on scores on the age and educated adjusted Folstein Mini-Mental Status Exam (MMSAdj) and a brief Delayed Verbal Recall Test (DRT). Subjects were assessed for APOE genotype, subjective memory complaints (Memory Questionnaire, MQ), depressive symptoms (Hamilton Depression Rating Scale, HDRS), and history of four major medical conditions that have been associated with memory loss (stroke/transient ischemic attack [TIA], atherosclerotic heart disease, hypertension, and diabetes). A hierarchical regression analysis was performed to examine the association between APOE genotype and memory complaints after controlling for a host of potential confounding factors. RESULTS: The APOE epsilon4 allele frequency for cognitively normal subjects was 0.13. Subjective memory complaints were predicted by depressive symptoms and a history of stroke/TIA. They were not associated with APOE genotype, MMSAdj score, DRT score, age, education, gender, and reported history of atherosclerotic heart disease, hypertension, or diabetes. CONCLUSION: The results did not suggest an association between subjective memory complaints and the APOE epsilon4 allele in this sample of cognitively intact subjects. This indicates that memory complaints may confer risk for future dementia through pathways independent of APOE genotype. The results also show that older adults with memory complaints are at increased risk for underlying depression. PMID- 15526314 TI - A simple hydrothermal route to large-scale synthesis of uniform silver nanowires. AB - This paper describes the preparation of uniform silver nanowires by reducing freshly prepared silver chloride with glucose at 180 degrees C for 18 hours in the absence of any surfactants or polymers. Scanning electron microscopy studies indicated that the silver nanowires are about 100 nm in diameter and up to 500 microm in length. High-resolution transmission electron microscopy analyses showed that the silver nanowires grow perpendicularly to the Ag(200) plane. The silver nanowires are believed to grow through a solid-solution-solid process. Some influential factors on the growth of silver nanowires are also discussed. PMID- 15526315 TI - Edge-bridging and face-capping coordination of alkenyl ligands in triruthenium carbonyl cluster complexes derived from hydrazines: synthetic, structural, theoretical, and kinetic studies. AB - The reactions of the triruthenium cluster complex [Ru3(mu-H)(mu3-eta2 HNNMe2)(CO)9] (1; H2NNMe2=1,1-dimethylhydrazine) with alkynes (PhC triple bond CPh, HC triple bond CH, MeO2CC triple bond CCO2Me, PhC triple bond CH, MeO2CC triple bond CH, HOMe2CC triple bond CH, 2-pyC triple bond CH) give trinuclear complexes containing edge-bridging and/or face-capping alkenyl ligands. Whereas the edge-bridged products are closed triangular species (three Ru-Ru bonds), the face-capped products are open derivatives (two Ru-Ru bonds). For terminal alkynes, products containing gem (RCCH2) and/or trans (RHCCH) alkenyl ligands have been identified in both edge-bridging and face-capping positions, except for the complex [Ru3(mu3-eta2-HNNMe2)(mu3-eta3-HCCH-2-py)(mu-CO)(CO)7], which has the two alkenyl H atoms in a cis arrangement. Under comparable reaction conditions (1:1 molar ratio, THF at reflux, time required for the consumption of complex 1), some reactions give a single product, but most give mixtures of isomers (not all the possible ones), which were separated. To determine the effect of the hydrazido ligand, the reactions of [Ru3(mu-H)(mu3-eta2-MeNNHMe)(CO)9] (2; HMeNNHMe=1,2-dimethylhydrazine) with PhC triple bond CPh, PhC triple bond CH, and HC triple bond CH were also studied. For edge-bridged alkenyl complexes, the Ru- Ru edge that is spanned by the alkenyl ligand depends on the position of the methyl groups on the hydrazido ligand. For face-capped alkenyl complexes, the relative orientation of the hydrazido and alkenyl ligands also depends on the position of the methyl groups on the hydrazido ligand. A kinetic analysis of the reaction of 1 with PhC[triple chemical bond]CPh revealed that the reaction follows an associative mechanism, which implies that incorporation of the alkyne in the cluster is rate-limiting and precedes the release of a CO ligand. X-ray diffraction, IR and NMR spectroscopy, and calculations of minimum-energy structures by DFT methods were used to characterize the products. A comparison of the absolute energies of isomeric compounds (obtained by DFT calculations) helped rationalize the experimental results. PMID- 15526316 TI - Synthesis of benzannulated N-heterocyclic carbene ligands by a template synthesis from 2-nitrophenyl isocyanide. AB - The reaction of 2-nitrophenyl isocyanide 2 with [M(CO)5(thf)] (M=Cr, Mo, W) yields the isocyanide complexes [M(CO)5(2)] (3: M=Cr; 4: M=Mo; 5: M=W). Complexes 3-5 react with elemental tin under reduction of the nitro function of the isocyanide ligand to give the complexes with the unstable 2-aminophenyl isocyanide ligand. The coordinated 2-aminophenyl isocyanide ligand in all three complexes reacts spontaneously under intramolecular nucleophilic attack of the primary amine at the isocyanide carbon atom to yield the complexes with the NH,NH benzimidazol-2-ylidene ligand (6: M=Cr; 7: M=Mo; 8: M=W). An incomplete reduction of the nitro group in 3-5 is observed when hydrazine hydrate is used instead of tin. Here the formation of complexes with a coordinated 2-hydroxylamine functionalized phenyl isocyanide [(CO)5M-CN-C6H(4-)-2-N(H)-OH] is postulated and this unstable ligand again undergoes intramolecular cyclization to give the NH,NOH-stabilized benzimidazol-2-ylidene complexes 9-11. The tungsten derivative 11 can be allylated stepwise by a deprotonation/alkylation sequence first at the OH and then at the NH position to yield the monoallylated and diallylated species 12 and 13. The molecular structures of 3-5 and 12-13 were established by X-ray crystallography. PMID- 15526317 TI - Oxorhenium complexes as aldehyde-olefination catalysts. AB - Several oxorhenium compounds in the formal oxidation states V and VII are examined as catalysts for the aldehyde-olefination starting from diazo compounds, phosphines, and aldehydes. Of these, [ReMeO2(eta2-alkyne)] complexes provide the simplest catalysts to study, although [ReOCl3(PPh3)2] still remains the most efficient rhenium catalyst for aldehyde-olefination described to date. Prior to the reaction with the Re catalysts the phosphine and the diazo compound react to form a phosphazine. No catalytic reaction occurs in cases where no phosphazine formation is observed. The first step of the catalytic cycle involves the formation of a carbene intermediate by the reaction of phosphazine and catalyst under extrusion of phosphine oxide and dinitrogen. In a second step the carbene reacts with aldehyde under olefin formation and catalyst regeneration. Excess of alkyne as well as the presence of ketones slows down the catalytic reaction. The olefination of 4-nitrobenzaldehyde with diazomalonate is possible with these Re catalysts. In contrast, this reaction does not take place either in the classical Wittig fashion from Ph3P=C(CO2Et)2 and aldehyde or by use of all other catalysts for aldehyde olefination reactions reported to date. Catalytic ylide formation from diazo compounds seems therefore not to be the only pathway through which catalytic aldehyde-olefination reactions can proceed. PMID- 15526318 TI - Solvent-ligated manganese(II) complexes for the homopolymerization of isobutene and the copolymerization of isobutene and isoprene. AB - Polyisobutenes with a high content of terminal olefinic groups can be synthesized by using manganese(II) initiators in homogeneous solution. These easily accessible complexes initiate the polymerization at room temperature and above, and afford highly reactive, gel-free polyisobutenes with high viscosities. Furthermore, the initiators were successfully used for the copolymerization of isobutene with isoprene. The high activities of the Mn(II) initiators seem to be related to their weakly coordinating nitrile ligands, which are easily displaced by substrate molecules. Replacing the nitrile ligands by other more strongly coordinating ligands such as water reduces the initiator activity significantly. The Mn(II) initiators are surprisingly resistant to temperature. PMID- 15526319 TI - Cytokines in pancreatic carcinoma: correlation with phenotypic characteristics and prognosis. AB - BACKGROUND: Cytokines have been implicated in diverse processes that are relevant to pancreatic carcinoma, including cachexia, asthenia, and tumor growth. The objective of this study was to examine the association between serum levels of proinflammatory and antiinflammatory or angiogenic cytokines and the outcomes of patients with pancreatic carcinoma. METHODS: Serum cytokine levels were measured by enzyme-linked immunosorbent assay from 51 patients with pancreatic carcinoma and from 48-62 healthy volunteers. Cytokine levels were compared with disease manifestations and overall survival. RESULTS: Circulating levels of vascular endothelial growth factor, tumor necrosis factor alpha, interleukin-1alpha (IL 1alpha), and IL-1beta were not elevated significantly in patients with pancreatic carcinoma, but levels of IL-6, IL-8, IL-10, and IL-1 receptor antagonist (IL-1RA) were elevated significantly (P <0.05). Cytokine levels were dichotomized based on an analysis of null Martingale residuals. Patients who had IL-6 levels > 5.2 pg/mL or IL-10 levels >9.8 pg/mL had significantly worse survival compared with patients who had lower IL-6 or IL-10 levels (P <0.05). IL-8 levels were not associated with survival differences. Patients who had IL-1RA levels <159 pg/mL had significantly worse survival compared with patients who had higher IL-1RA levels (P <0.05). Higher IL-6, IL-10, and IL-8 levels were associated with poor performance status and/or weight loss. In multivariate analysis, only T4 tumors and high IL-6 levels were selected as independent prognostic factors for poor survival. CONCLUSIONS: Circulating levels of several cytokines were high in patients with pancreatic carcinoma, and their association with weight loss and poor performance status suggested that they may be involved in these disease manifestations. Furthermore, serum cytokine levels, in particular IL-6, may be a useful prognostic marker. PMID- 15526320 TI - Synthesis, spectroscopy, and electrochemistry of tetra-tert-butylated tetraazaporphyrins, phthalocyanines, naphthalocyanines, and anthracocyanines, together with molecular orbital calculations. AB - Tetraazaporphyrins (TAPs), phthalocyanines (Pcs), naphthalocyanines (Ncs), and anthracocyanines (Acs) with four tert-butyl groups attached at similar positions have been synthesized, and their electronic absorption, magnetic circular dichroism (MCD), IR, and voltammetric properties were studied and interpreted with the help of quantum-mechanical calculations. Through the preparation of a series of compounds with the same number of the same substituent, the effects of the increase in the size of the ring system were clearly derived. The main results may be summarized as follows. 1) The Q band shifts to longer wavelength and its intensity increases, but with decreasing degree of change with increasing molecular size. If the size of the effect of benzene directly fused to the TAP skeleton is set at unity, the effects of the second and third benzene units are roughly 0.8 and 0.5, respectively. 2) The splitting of the Q bands in metal-free compounds decreases with increasing molecular size, so that the Q bands of H2Nc and H2Ac appear as single bands. 3) The magnitude of the orbital angular momentum of the excited state of the ligand decreases with increasing molecular size. 4) Interestingly, the ring current, as judged from the positions of pyrrole proton signals in the 1H NMR spectrum, appears to decrease with increasing molecular size. 5) The first reduction potential becomes less negative, but only slightly, whereas the first oxidation potential shows a marked shift to less positive values with increasing molecular size, indicating that the HOMO destabilizes significantly as the molecule becomes larger. 6) In 5), the extent of the HOMO destabilization with molecular size differs depending on the central metal, so metals producing smaller destabilization effects can allow larger macrocycles. Of the metals studied, the most effective is cobalt, and the practical size limit is represented by the Acs. 7) The IR spectra become simpler the larger the molecule, and the main bands were assigned by DFT calculations. 8) The trend in experimentally determined redox potentials and electronic absorption and MCD spectra were reasonably reproduced by MO calculations using the ZINDO/S Hamiltonian. 9) EPR data for several metallocomplexes are also reported. PMID- 15526321 TI - Trends in mortality from major cancers in the European Union, including acceding countries, in 2004. AB - BACKGROUND: In May 2004, 10 additional countries joined the European Union (EU), including a total of 75 million inhabitants. Most of these were from central and eastern European countries with comparably high cancer mortality rates and with relatively unfavorable trends. Therefore, it is important to provide updated mortality data regarding major cancers in various countries and to analyze trends for the current population of the EU. METHODS: The authors considered mortality rates (directly standardized to the world standard population) for all cancers and for 8 major cancer sites in the year 2000 in the 25 countries of the EU and analyzed corresponding trends since 1980 using data derived from the World Health Organization data base. RESULTS: For men, overall cancer mortality in the year 2000 varied by a factor > 2 between the highest rate of 258.5 per 100,000 men in Hungary and the lowest rate of 122.0 per 100,000 men in Sweden. Central and Eastern European accession countries had the highest rates not only for lung and other tobacco-related cancers but also for gastrointestinal cancers and leukemias. The geographic pattern was different and the range of variation was smaller for women, i.e., between 136.7 per 100,000 women in Denmark and 76.4 per 100,000 women in Spain in the year 2000. In the EU as a whole, lung cancer mortality in men peaked at 55.4 per 100,000 men in 1988 and declined thereafter to 46.7 per 100,000 men in 2000. Gastric cancer steadily declined from 19.7 per 100,000 men in 1980 to 10.1 per 100,000 men in 2000. Other major sites showed moderately favorable trends over the last few years. In women, breast cancer peaked at 21.7 per 100,000 in 1989 and declined to 18.9 per 100,000 in 2000. Mortality from gastric, (cervix) uterus, and intestinal cancers demonstrated steady decreases, but lung cancer increased from 7.7 per 100,000 women in 1980 to 11.1 per 100,000 women in 2000. The increase in lung cancer mortality in women age < 55 years was 38% between 1990 and 2000 (from 2.16 per 100,000 women to 2.99 per 100,000 women), reflecting the spread of tobacco smoking among women in the EU over the last few decades. CONCLUSIONS: The priority for further reduction of cancer mortality in the EU remains tobacco control together with more widespread availability of modern diagnostic and treatment procedures for neoplasms that are amenable to treatment. PMID- 15526322 TI - Colorectal carcinoma mortality among Appalachian men and women, 1969-1999. AB - BACKGROUND: Colorectal carcinoma screening can reduce mortality, but residents of poor or medically underserved areas may face barriers to screening. The current study assessed colorectal carcinoma mortality in Appalachia, a historically underserved area, from 1969 to 1999. METHODS: All counties within the 13-state Appalachian region, which stretches from southern New York to northern Mississippi, were used to calculate annual death rates for the 31-year period. Joinpoint regression analysis was used to examine trends by age and race for the Appalachian region and the remainder of the United States. Five-year rates for 1995-1999 age-adjusted to the 2000 U.S. standard population were calculated by race and age group for the Appalachian region and elsewhere in the United States. RESULTS: Trend analysis showed that colorectal carcinoma death rates among both racial and gender groups studied had declined in recent years. Despite this, the rates for white males and white females were still significantly higher in Appalachia than in the rest of the country at the end of the study period, 1999. Five-year colorectal carcinoma death rates among white males (ages < 50, 50-59, and 70-79 years) and white females (ages < 50, 50-59, 70-79, > or = 80 years) were significantly higher in Appalachia than elsewhere in the United States, whereas rates among black females 60-69 and 70-79 years old were significantly lower in Appalachia. CONCLUSIONS: The Appalachian region may benefit from targeted prevention efforts to eliminate disparities in the colorectal carcinoma death rates among subgroups. Further studies are needed to determine whether the higher death rates in specific Appalachian subgroups are related to a higher incidence of the disease, the cancer being at a later stage at diagnosis, poorer treatment, or other factors. PMID- 15526323 TI - Method and algorithm of obtaining the molecular intrinsic characteristic contours (MICCs) of organic molecules. AB - The molecular intrinsic characteristic contour (MICC) is defined as the set of all the classical turning points of electron movement in a molecule. Studies on the MICCs of some medium organic molecules, such as dimethylether, acetone, and some homologues of alkanes, alkenes, and alkynes, as well as the electron density distributions on the MICCs, are shown for the first time. Results show that the MICC is an intrinsic approach to shape and size of a molecule. Unlike the van der Waals hard-sphere model, the MICC is a smooth contour, and it has a clear physical meaning. Detailed investigations on the cross-sections of MICCs have provided a kind of important information about atomic size changing in the process of forming molecules. Studies on electron density distribution on the MICC not only provide a new insight into molecular shape, but also show that the electron density distribution on the boundary surface relates closely with molecular properties and reactivities. For the homologues of alkanes, Rout(H), Dmin, and Dmax (the minimum and maximum of electron density on the MICC), all have very good linear relationships with minus of the molecular ionization potential. This work may serve as a basis for exploring a new reactivity indicator of chemical reactions and for studying molecular shape properties of large organic and biological molecules. PMID- 15526325 TI - Evaluation of library ranking efficacy in virtual screening. AB - We present the results of a comprehensive study in which we explored how the docking procedure affects the performance of a virtual screening approach. We used four docking engines and applied 10 scoring functions to the top-ranked docking solutions of seeded databases against six target proteins. The scores of the experimental poses were placed within the total set to assess whether the scoring function required an accurate pose to provide the appropriate rank for the seeded compounds. This method allows a direct comparison of library ranking efficacy. Our results indicate that the LigandFit/Ligscore1 and LigandFit/GOLD docking/scoring combinations, and to a lesser degree FlexX/FlexX, Glide/Ligscore1, DOCK/PMF (Tripos implementation), LigandFit1/Ligscore2 and LigandFit/PMF (Tripos implementation) were able to retrieve the highest number of actives at a 10% fraction of the database when all targets were looked upon collectively. We also show that the scoring functions rank the observed binding modes higher than the inaccurate poses provided that the experimental poses are available. This finding stresses the discriminatory ability of the scoring algorithms, when better poses are available, and suggests that the number of false positives can be lowered with conformers closer to bioactive ones. PMID- 15526326 TI - QMQSAR: utilization of a semiempirical probe potential in a field-based QSAR method. AB - A semiempirical quantum mechanical approach is described for the creation of molecular field-based QSAR models from a set of aligned ligand structures. Each ligand is characterized by a set of probe interaction energy (PIE) values computed at various grid points located near the surface of the ligand. Single point PM3 calculations afford these PIE values, which represents a pool of independent variables from which multilinear regression models of activity are built. The best n-variable fit is determined by constructing an initial regression using standard forward stepwise selection, followed by refinement using a simulated annealing technique. The resulting fit provides an easily interpreted 3D physical model of ligand binding affinity. Validation against three literature datasets demonstrates the ability of the semiempirical potential to model critical binding interactions in diverse systems. PMID- 15526327 TI - Novel activators of the tumour suppressor p53. PMID- 15526328 TI - A miniature membrane-less biofuel cell operating at +0.60 V under physiological conditions. PMID- 15526329 TI - Effect of silver nanoparticles on the electron transfer reactivity and the catalytic activity of myoglobin. AB - Silver nanoparticles (11+/-1.5 nm) could greatly enhance the electron-transfer reactivity of myoglobin (Mb) and its catalytic ability toward hydrogen peroxide (H2O2). Direct fast electron transfer between Mb and a pyrolytic graphite (PG) electrode was achieved, and a pair of well-defined, quasi-reversible redox peaks was obtained. The cathodic and anodic peaks were located at -329 and -281 mV, respectively. Meanwhile, the catalytic ability of the protein toward the reduction of H2O2 was also studied, and a H2O2 biosensor was subsequently fabricated. Its detection limit was 1.0 x 10(-6) M with a sensitivity of 0.0205 microA per microM of H2O2. The apparent Michaelis-Menten constant was calculated to be 1303 muM. Flocculation assay showed that the protein maintained plasmon layers surrounding the surface of silver nanoparticles and avoided silver nanoparticle aggregation. On the other hand, UV-visible spectroscopy studies revealed that silver nanoparticles could induce a small change of the heme-group environment of the protein; this contributed to the enhancement of the electron transfer reactivity and the catalytic activity. PMID- 15526330 TI - Structural investigation of the HIV-1 envelope glycoprotein gp160 cleavage site 3: role of site-specific mutations. AB - Proteolytic processing of HIV gp160 to produce gp120 and gp41 is performed by PC enzymes. This process is a prerequisite for the virus infectivity, since both gp120 and gp41 participate in the virus HIV-1 entry mechanism. The structure of the gp120/gp41 junction remains to be elucidated, and the structural features required for molecular recognition between HIV-1 gp160 and proteolytic enzymes have not been clarified. Furin is the best PC candidate for the gp160 proteolytic processing known to date. In previous studies on model peptides, we have shown the relevance of an N-terminal helix for the proper recognition of the gp160 processing site by furin. Here we analyze the effect of point mutations in peptides lacking a regular N-terminal helix. To this end, we present the structure-activity characterization of three peptide analogues of the HIV gp160 processing site that all present mutations in proline at positions P3 and/or P2', while sharing the same N-terminal sequence, containing helix-breaking D-amino acids. Conformational analysis of the peptides was carried out in solution by NMR techniques, and furin's efficiency in cleaving them was measured. Structural findings are presented and discussed in relation to the different exhibited activity. PMID- 15526332 TI - Real-time monitoring of in vitro transcriptional RNA by using fluorescence correlation spectroscopy. PMID- 15526331 TI - Structure and axis curvature in two dA6 x dT6 DNA oligonucleotides: comparison of molecular dynamics simulations with results from crystallography and NMR spectroscopy. AB - Molecular dynamics (MD) simulations have been performed on the A6 containing DNA dodecamers d(GGCAAAAAACGG) solved by NMR and d(CGCAAAAAAGCG) solved by crystallography. The experimental structures differ in the direction of axis bending and in other small but important aspects relevant to the DNA curvature problem. Five nanosecond MD simulations of each sequence have been performed, beginning with both the NMR and crystal forms as well as canonical B-form DNA. The results show that all simulations converge to a common form in close proximity to the observed NMR structure, indicating that the structure obtained in the crystal is likely a strained form due to packing effects. A-tracts in the MD model are essentially straight. The origin of axis curvature is found at pyrimidine-purine steps in the flanking sequences. PMID- 15526333 TI - A molecular dynamics study of acylphosphatase in aggregation-promoting conditions: the influence of trifluoroethanol/water solvent. AB - The 98-residue protein acylphosphatase exhibits a high propensity for aggregation under certain conditions. Aggregates formed from wild-type acylphosphatase in the presence of 2,2,2-trifluoroethanol and from highly destabilized mutants are essentially identical in structure. Furthermore, it has been shown by mutational studies that different regions of the protein are important for aggregation and folding. In the present molecular dynamics study, we compare the behavior of the protein in aqueous solution and in a 25% (v/v) 2,2,2-trifluoroethanol/water environment mimicking the experimental conditions. The 2,2,2-trifluoroethanol surrounding affects the structure of the protein mostly in the regions important for aggregation, in good agreement with experimental data. This suggests that the early step of (partly) unfolding, which precedes the aggregation process, has been observed. PMID- 15526334 TI - Peptide-chelating agent conjugate for selective targeting of somatostatin receptor type 1: synthesis and characterization. AB - Previously reported results suggest that the analogue of the somatostatin des AA1,2,5[D-Trp8,IAmp9]-somatostatin (CH-275) peptide bearing chelating agents able to coordinate radioactive metals could be used for scintigraphic imaging of tumor lesions overexpressing sstr1. An efficient synthetic procedure for the preparation of the somatostatin analogue CH-275 and its conjugate DTPAGlu-Gly-CH 275, bearing the chelating agent DTPAGlu (DTPAGlu=N,N-bis[2-[bis(carboxy ethyl)amino]ethyl]-L-glutamic acid) on the N-terminus, by solid-phase peptide synthesis and 9-fluorenylmethoxycarbonyl (Fmoc) chemistry, is here reported. Rapid and efficient labeling of DTPAGlu-Gly-CH-275 was achieved by addition of 111In(III) to the compound. Typical yields were greater than 97% as determined by reversed phase high performance liquid chromatography (HPLC) at specific activities in the range 4-9 GBq/micromol (100-250 Ci/mmol). A preliminary biological assay of the binding ability of 111In-DTPAGlu-Gly-CH-275 indicates, however, that the labeled compound does not display any specific interaction with somatostatin sstr1 receptors in the tested cell lines. To confirm this unexpected negative result, competition binding experiments were carried out, in which fixed tracer amounts of the 125I-labeled somatostatin-14 were incubated with the receptor-expressing cells in the presence of DTPAGlu-Gly-CH-275 or CH-275 at concentrations ranging from 10(-10) to 10(-3) M. While CH-275 shows a IC50 of 80 nM similar to that already found in displacement experiments on CHO-K1 sstr1 transfected cells, DTPAGlu-Gly-CH-275 displays instead very low or negligible affinity towards this receptor. The NMR solution characterization indicates that the presence of DTPAGlu does not influence the conformational and chemical features of the peptide moiety, thus suggesting that the loss in binding activity should be due to steric hindrance of either the chelating agent DTPAGlu or its indium complex. PMID- 15526335 TI - Polymyxin B-lipid interactions in Langmuir-Blodgett monolayers of Escherichia coli lipids: a thermodynamic and atomic force microscopy study. AB - The dramatically increased frequency of antibiotic resistance has led to intensive efforts towards developing new families of antibiotics. Membrane-active antibiotic peptides such as polymyxin B (PxB) hold promise as the next generation of antibiotics, since they rarely spur the evolution of resistance. At low concentrations in the membrane, PxB forms vesicle-vesicle contacts and induces lipid exchange without leakage or fusion, a phenomenon that can explain its specificity towards gram-negative bacteria by contact formation between the two phospholipids interfaces in the periplasmatic space. In this work, the interaction of PxB and the nonantibiotic derivative polymyxin B nonapeptide (PxB NP) with monolayers of Escherichia coli membrane lipids (ECL) has been studied by thermodynamic and structural methods. PxB inserts itself into ECL monolayers as a conformation that forms intermembrane contacts with vesicles injected underneath, and induces lipid exchange when the monolayer surface pressure is set at 32 mN/m (membrane equivalence pressure) or net transfer vesicle-to-monolayer at lower surface pressures. Thermodynamic analysis of the compression isotherms of mixed monolayers indicates that PxB inserts into the monolayer with an expansion of the mean molecular area, implying that peptide and lipids form nonideal mixtures. At low concentrations, corresponding to the membrane-membrane contact form of PxB, the mixed monolayers present positive excess energy values (deltaGm(Ex)), and atomic force microscopy (AFM) imaging reveals structures of approximately 120-nm diameter that protrude from the lipid surface approximately 0.7 nm. At concentrations of PxB above 4 mol %, thermodynamic analysis gives a very high deltaGm(Ex), corresponding to nonfavorable interactions, and AFM images show round structures of 20-30 nm diameter. PxB-NP behaves in a totally different way, in agreement with its inability to form vesicle-vesicle contacts and its lack of antibiotic effect. These results are discussed in the light of the mechanism of action of PxB on the membrane of gram-negative bacteria. PMID- 15526336 TI - New synthesis of PNA-3'DNA linker monomers, useful building blocks to obtain PNA/DNA chimeras. AB - A new synthetic strategy to get the PNA-3'DNA linker with the monomethoxytrityl (Mmt) group as temporary protection of the backbone to be used for the synthesis of PNA/DNA chimeras was employed and a convenient strategy to obtain Mmt PNA monomers was developed. The synthetic strategies take advantage of the introduction of the acid-labile Mmt-protecting group in the first step. PMID- 15526337 TI - Change in enantioselectivity in bufuralol 1''-hydroxylation by the substitution of phenylalanine-120 by alanine in cytochrome P450 2D6. AB - The functional roles of phenylalanine at position 120 in drug oxidation by cytochrome P450 2D6 (CYP2D6) were examined using a yeast cell expression system and bufuralol (BF) enantiomers as a chiral substrate. Two mutated cDNAs, one encoding a CYP2D6 mutant having alanine instead of Phe-120 (F120A) and another encoding a mutant having alanine instead of Glu-222 (E222A), were prepared by site-directed mutagenesis and transformed into yeast cells via pGYRI vectors. The enantiomeric BF 1''-hydroxylase activities of the mutants were compared with those of the wild type. When enantiomeric BF 1''-hydroxylase activities at a substrate concentration of 100 microM were compared, the CYP2D6 wild type showed substrate enantioselectivity of (R-BF >> S-BF) and the F120A mutant exhibited substrate enantioselectivity of (R-BF < or = S-BF), whereas the product diastereoselectivity of (1''R-OH-BF << 1''-S-OH-BF) was similar between the wild type and the mutant. The activities of the other mutant (E222A) were much lower than those of the wild type and the F120A mutant, while its substrate enantioselectivity and product diastereoselectivity were the same as those of the wild type. The kinetics demonstrated that apparent K(m) values were similar among the recombinant enzymes, and V(max) values clearly reflected the selectivity described above. These results indicate that Phe-120 has a key role in the enantioselective BF 1''-hydroxylation by CYP2D6. PMID- 15526338 TI - Synthesis of new N-sulfonylated amino alcohols and application to the enantioselective addition of diethylzinc to aldehydes. AB - A series of N-sulfonylated amino alcohols were prepared and the enantioselective addition of diethylzinc to aldehydes catalyzed by titanium(IV) complexes of those ligands were carried out. Excellent enantioselectivities were obtained for the desired alcohols with enantiomeric excess values up to 94% of R-configuration. PMID- 15526339 TI - One-pot synthesis and chiral analysis of cyclopropane derivatives. AB - A user-friendly, one-pot procedure was developed to access racemic as well as enantiomerically enriched cyclopropanes. Thus, the cyclopropanation of olefin (3) was performed using Meldrum's acid (4) or dimethyl malonate (5) and diacetoxyiodobenzene PhI(OAc)2 (6) or iodosyl benzene PhI=O (7) for in situ generation and decomposition of the phenyliodonium ylide 1 and 2, respectively. The reaction proceeds well with 5 mol% of achiral rhodium (II)-catalyst [Rh2(OAc)4] and a 10-fold excess of olefin affording the cyclopropane derivates 10 and 11, respectively, with high yield. The system is compatible with chiral Rh(II)-catalysts 8 and 9 and an enantiomeric excess up to 66% was achieved. An effective baseline separation of the enantiomers of the resulting cyclopropane derivatives was achieved using gas chromatography on the chiral stationary phase Chirasil-beta-dex. PMID- 15526340 TI - Enantioselective synthesis of 2- and 3-substituted 2,3-dihydro[1,4]dioxino[2,3 b]pyridine derivatives and enantiomeric purity control by capillary electrophoresis. AB - A rapid and simple procedure for enantioselective preparation of 2- and 3 substituted 2,3-dihydro[1,4]dioxino[2,3-b]pyridine derivatives (A and B, respectively) is described. The enantiomeric purity of each isomer was determined by capillary electrophoresis using a dual-cyclodextrin system (S-beta-CD/beta-CD) dissolved in formic acid-ammonia buffer (pH 4, ionic strength 50 mM). PMID- 15526341 TI - Molybdenum phosphide as an o-propylaniline hydrodenitrogenation catalyst: a first principles study. AB - MoP has been shown experimentally to be an active catalyst in the hydrodenitrogenation of o-propylaniline. We investigate the structure and the energetics of the o-propylaniline adsorption on the Mo-terminated MoP(001) surface. Detailed information on the structure of the free MoP(001) surface and on the structure and adsorption energy of o-propylaniline on MoP(001) is obtained by using density functional theory. The transition state, reaction path, and the energy barrier are reported for one of the branches of the HDN reaction network that leads to the formation of propylbenzene by hydrogenolysis of the C-N bond. PMID- 15526342 TI - Palladium-catalyzed intramolecular hydroalkylation of alkenyl-beta-keto esters, alpha-aryl ketones, and alkyl ketones in the presence of Me3SiCl or HCl. AB - Reaction of 3-butenyl beta-keto esters or 3-butenyl alpha-aryl ketones with a catalytic amount of [PdCl2(CH3CN)2] (2) and a stoichiometric amount of Me3SiCl or Me3SiCl/CuCl2 in dioxane at 25-70 degrees C formed 2-substituted cyclohexanones in good yield with high regioselectivity. This protocol tolerated a number of ester and aryl groups and tolerated substitution at the allylic, enolic, and cis and trans terminal olefinic positions. In situ NMR experiments indicated that the chlorosilane was not directly involved in palladium-catalyzed hydroalkylation, but rather served as a source of HCl, which presumably catalyzes enolization of the ketone. Identification of HCl as the active promoter of palladium-catalyzed hydroalkylation led to the development of an effective protocol for the hydroalkylation of alkyl 3-butenyl ketones that employed sub-stoichiometric amounts of 2, HCl, and CuCl2 in a sealed tube at 70 degrees C. PMID- 15526343 TI - A proteomic analysis of thioacetamide-induced hepatotoxicity and cirrhosis in rat livers. AB - Thioacetamide (TAA) administration is an established technique for generating rat models of liver fibrosis and cirrhosis. Oxidative stress is believed to be involved as TAA-induced liver fibrosis is initiated by thioacetamide S-oxide, which is derived from the biotransformation of TAA by the microsomal flavine adenine dinucleotide (FAD)-containing monooxygense (FMO) and cytochrome P450 systems. A two-dimensional gel electrophoresis-mass spectrometry approach was applied to analyze the protein profiles of livers of rats administered with sublethal doses of TAA for 3, 6 and 10 weeks respectively. With this approach, 59 protein spots whose expression levels changed significantly upon TAA administration were identified, including three novel proteins. These proteins were then sorted according to their common biochemical properties and functions, so that pathways involved in the pathogenesis of rat liver fibrosis due to TAA induced toxicity could be elucidated. As a result, it was found that TAA administration down-regulated the enzymes of the primary metabolic pathways such as fatty acid beta-oxidation, branched chain amino acids and methionine breakdown. This phenomenon is suggestive of the depletion of succinyl-CoA which affects heme and iron metabolism. Up-regulated proteins, on the other hand, are related to oxidative stress and lipid peroxidation. Finally, these proteomics data and the data obtained from the scientific literature were integrated into an "overview model" for TAA-induced liver cirrhosis. This model could now serve as a useful resource for researchers working in the same area. PMID- 15526344 TI - Differential expression profiling of human pancreatic adenocarcinoma and healthy pancreatic tissue. AB - Due to poor prognosis and lack of effective treatment, pancreatic carcinoma (PC) is a devastating disease. With the goal of contributing to an improved detection, prevention and treatment of the disease, a comparative proteome analysis of PC and normal tissue was carried out. Paired tissue extracts from 12 patients (pancreatic adenocarcinoma and adjacent healthy tissue) were separated by two dimensional electrophoresis. Differential protein expression was analyzed by gel comparison with the help of image analysis software. The differentially expressed spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Seventy proteins were more strongly expressed (mostly two-fold or more) in cancerous tissue, while 41 were stronger in normal pancreas respectively. Those spots highly expressed in PC were confirmed in gels from independent individual samples. Among them were several cytoskeletal proteins, small GTP-binding proteins, and members of the S100 protein family etc. Nine proteins had been reported in previous nuclear acid-based studies. The levels of two proteins were confirmed by immunohistochemistry. One of them, fascin, was detected in 13 out of 21 carcinoma and negative in all normal pancreas samples. Moreover, fascin expression was related to the differentiation of pancreatic carcinoma. PMID- 15526345 TI - Proteome analysis of human substantia nigra in Parkinson's disease. AB - Protein expression has been compared in human substantia nigra specimens from Parkinson's disease (PD) patients and from controls, and 44 proteins expressed in this midbrain region were identified by peptide mass fingerprinting. Among them, nine showed changes in their abundance. L and M neurofilament chains are less abundant in PD specimens, whereas peroxiredoxin II, mitochondrial complex III, ATP synthase D chain, complexin I, profilin, L-type calcium channel delta subunit, and fatty-acid binding protein are significantly more present in PD samples than in controls. Besides the consolidated view of oxidative stress involvement in PD pathogenesis, suggested by overexpression of mitochondrial and reactive oxygen species (ROS)-scavenging proteins, these results indicate a possible potentiation mechanism of afferent signals to substantia nigra following degeneration of dopaminergic neurons. PMID- 15526346 TI - Characterization of the in vivo forms of lacrimal-specific proline-rich proteins in human tear fluid. AB - The tear film is complex and is rich in both peptides and proteins. Physiological factors have been shown to alter the balance of the protein components in the tear film, however, little is known of the precise stimuli that initiate these changes, or their nature and extent. Attention has been directed at the role of tear proteins in the protection of the external ocular surface, and their potential role in the pathogenesis of inflammatory and autoimmune diseases, but few lacrimal-specific proteins have been identified and demonstrated to offer a protective function at the ocular surface. The biological importance of proline rich proteins is uncertain, although there is some evidence to indicate a potential antimicrobial function for these proteins in saliva. Despite the detection of mRNA for proline-rich proteins in lacrimal gland, the translated protein product has not been detected in tear fluid. In this study we investigate the presence of proline-rich proteins in the tear film. Human reflex tear fluid was examined by matrix-assisted laser desorption/ionization-time of flight mass spectrometry directly, and following size exclusion high performance liquid chromatography. This revealed significant levels of a truncated form of lacrimal proline-rich protein, and a series of peptides derived the C-terminus of this protein. None of these had previously been identified in tear. Our study highlights the dangers inherent in proteomic strategies that assign an identity to a protein based on limited coverage of tryptic peptides. PMID- 15526347 TI - Iron oxide MR contrast agents for molecular and cellular imaging. AB - Molecular and cellular MR imaging is a rapidly growing field that aims to visualize targeted macromolecules or cells in living organisms. In order to provide a different signal intensity of the target, gadolinium-based MR contrast agents can be employed although they suffer from an inherent high threshold of detectability. Superparamagnetic iron oxide (SPIO) particles can be detected at micromolar concentrations of iron, and offer sufficient sensitivity for T2(*) weighted imaging. Over the past two decades, biocompatible particles have been linked to specific ligands for molecular imaging. However, due to their relatively large size and clearance by the reticuloendothelial system (RES), widespread biomedical molecular applications have yet to be implemented and few studies have been reproduced between different laboratories. SPIO-based cellular imaging, on the other hand, has now become an established technique to label and detect the cells of interest. Imaging of macrophage activity was the initial and still is the most significant application, in particular for tumor staging of the liver and lymph nodes, with several products either approved or in clinical trials. The ability to now also label non-phagocytic cells in culture using derivatized particles, followed by transplantation or transfusion in living organisms, has led to an active research interest to monitor the cellular biodistribution in vivo including cell migration and trafficking. While most of these studies to date have been mere of the 'proof-of-principle' type, further exploitation of this technique will be aimed at obtaining a deeper insight into the dynamics of in vivo cell biology, including lymphocyte trafficking, and at monitoring therapies that are based on the use of stem cells and progenitors. PMID- 15526348 TI - Feridex labeling of mesenchymal stem cells inhibits chondrogenesis but not adipogenesis or osteogenesis. AB - Magnetic resonance (MR) tracking of superparamagnetic iron oxide (SPIO)-labeled cells is a relatively new technique to non-invasively determine the biodistribution and migration of transplanted stem cells. A number of studies have recently reported encouraging results in the use of bone marrow-derived mesenchymal stem cells (MSCs) for repair of a variety of tissues. For MR tracking of SPIO-labeled MSCs, it is important to determine the effect that the magnetic labeling procedure may have on the differentiation capacity of labeled MSCs. Human MSCs were labeled with poly-L-lysine (PLL)-coated Feridex, with Feridex being an FDA-approved SPIO formulation in an off-label application, and assayed for cellular differentiation using five different assays. As compared with unlabeled controls, labeled MSCs exhibited an unaltered viability, proliferated similarly, and underwent normal adipogenic and osteogenic differentiation. However, there was a marked inhibition of chondrogenesis. The blocking of chondrogenic activity was mediated by the Feridex, rather than by the transfection agent (PLL). This is the first report showing Feridex blocking of cellular differentiation down a specific pathway (while not affecting viability and proliferation), and caution should thus be exercised when using Feridex labeled MSCs for chondrogenic MR tracking studies. On the other hand, no detrimental effects of Feridex-labeling are anticipated for MR-guided osteogenic or adipogenic transplantation studies. PMID- 15526349 TI - Applications of ultrasmall superparamagnetic iron oxide contrast agents in the MR study of animal models. AB - Ultrasmall superparamagnetic iron oxide nanoparticles have been widely used during the past decade as MR intravascular contrast agents in the study of animal models. Such agents enhance both T1 and T2/T2* relaxation, although for animal studies it is the later type of enhancement that is most commonly exploited. Their strong microscopic intravascular susceptibility effect enables the local blood volume distribution to be mapped in various organs. High spatial resolution and sensitivity can be achieved, because the long half-life of these agents in blood, combined with anesthetization, permits steady-state measurements over extended periods. This capability has been utilized to study the cerebrovascular blood volume distributions and their changes in normal, activated, pathologic and pharmacologically or genetically modified states, particularly in rodent animal models. It has also been applied to study blood volume changes in other tissues, such as the myocardium. The relaxation rate shifts Delta R2 and Delta R2* induced by iron oxide agents may differ depending on certain morphological characteristics of the microvascular network, and sensitive Delta R2 and Delta R2* mapping can potentially provide, in addition to blood volume, measurement of other important microvascular parameters such as blood vessel density and size. This work aims to review the applications of ultrasmall superparamagnetic iron oxide contrast agents in MR animal studies, with an emphasis on the investigation of microvascular parameters. PMID- 15526350 TI - Hepatocyte growth factor (HGF) downregulates thrombospondin 1 (TSP-1) expression in thyroid papillary carcinoma cells. AB - This study investigates the expression of thrombospondin-1 in papillary carcinoma of the thyroid and the role of Met-HGF interaction in TSP-1 regulation. In tissue sections, immunostaining for TSP-1 was associated with the fibrous tumour stroma, and showed areas of marked intensity adjacent to the basal membrane of tumour cells. Investigation of TSP-1 RNA expression showed that, in 10 of 14 cases, TSP 1 mRNA levels were significantly lower in tumour tissue (20-100% reduction; mean = 55% +/- 20; p = 0.001) than in the corresponding normal thyroid. Since it has been reported that HGF can downregulate the expression of TSP-1 mRNA, TSP-1 mRNA levels were measured in 7 primary cultures, established from thyroid papillary carcinomas (TPC), and in 1 TPC cell line prior to, or after, stimulation with HGF. A marked decrease in TSP-1 mRNA levels was observed after HGF stimulation in 6/7 primary cultures (60-100% decrease (mean = 79 +/- 15%; p = 0.006) and in the TPC cell line; moreover, the decrease in TSP-1 mRNA in cell extracts was associated with a decrease in TSP-1 protein in culture supernatants. The HGF activity was dose dependent and the downregulation lasted for at least 48 h after stimulation. The high-level expression of Met protein, the high-affinity receptor for HGF, in most cases of papillary carcinoma of the thyroid is consistent with the possibility that HGF-Met interaction plays a crucial role in regulating the expression of TSP-1 in this tumour type. PMID- 15526351 TI - The utility of superparamagnetic contrast agents in MRI: theoretical consideration and applications in the cardiovascular system. AB - This review will discuss the in vivo physical chemical relaxation properties of superparamagnetic iron oxide particles. Various parameters such as size, magnetization, compartmentalization and water exchange effects and how these alter the behavior of the iron oxide particles in an in vitro vs an in vivo situation with special reference to the cardiovascular system will be exemplified. Furthermore, applications using iron oxide particles for vascular, perfusion and viability imaging as well as assessment of the inflammatory status of a given tissue will be discussed. PMID- 15526352 TI - Relaxation induced by ferritin: a better understanding for an improved MRI iron quantification. AB - Ferritin, the iron storing protein, is known to darken T2-weighted MRI. This darkening can be used to non-invasively measure iron content. However, ferritin's behavior is not the same in tissue as in solution, a discrepancy that remains unexplained by the recently developed theory matching the NMR properties of ferritin solutions. A better understanding of the relaxation induced by ferritin in tissue could help for the development of new MRI protocols of iron quantification. In this short review, the main relaxation properties of ferritin in solution and in tissue are presented together with a discussion of the possible reasons for the faster transverse relaxation observed in tissues. PMID- 15526353 TI - GSTM1, GSTT1, GSTP1 and CYP1A1 genetic polymorphisms and susceptibility to esophageal cancer in a French population: different pattern of squamous cell carcinoma and adenocarcinoma. AB - AIM: To evaluate the association between CYP1A1 and GSTs genetic polymorphisms and susceptibility to esophageal squamous cell carcinoma (SCC) and esophageal adenocarcinoma (ADC) in a high risk area of northwest of France. METHODS: A case control study was conducted to investigate the genetic polymorphisms of these enzymes (CYP1A1*2C and GSTP1 exon 7 Val alleles, GSTM1*2/*2 and GSTT1*2/*2 null genotypes). A total of 79 esophageal cancer cases and 130 controls were recruited. RESULTS: GSTM1*2/*2 and CYP1A1*1A/*2C genotype frequencies were higher among squamous cell carcinomas at a level close to statistical significance (OR = 1.83, 95% CI 0.88-3.83, P = 0.11; OR = 3.03, 95% CI 0.93-9.90, P = 0.07, respectively). For GSTP1 polymorphism, no difference was found between controls and cases, whatever their histological status. Lower frequency of GSTT1 deletion was observed in ADC group compared to controls with a statistically significant difference (OR = 13.31, 95% CI 1.66-106.92, P<0.01). CONCLUSION: In SCC, our results are consistent with the strong association of this kind of tumour with tobacco exposure. In ADC, our results suggest 3 distinct hypotheses: (1) activation of exogenous procarcinogens, such as small halogenated compounds by GSTT1; (2) contribution of GSTT1 to the inflammatory response of esophageal mucosa, which is known to be a strong risk factor for ADC, possibly through leukotriene synthesis; (3) higher sensitivity to the inflammatory process associated with intracellular depletion of glutathione. PMID- 15526354 TI - Expression of Dnmt1, demethylase, MeCP2 and methylation of tumor-related genes in human gastric cancer. AB - AIM: To explore the effect of DNA methyltransferase, demethylase and methyl-CpG binding protein MeCP2 on the expressions and methylation of hMSH2 and proto oncogene in human gastric cancer. METHODS: Paired samples of primary gastric cancer and corresponding para-cancerous, non-cancerous gastric mucosae were obtained from surgically resected specimens of 28 patients. Transcription levels of Dnmt1, mbd2, MeCP2, p16(INK4A), hMSH2 and c-myc were detected by using real time PCR or RT-PCR. Promoter methylation of p16(INK4A), c-myc and hMSH2 genes was assayed by methylation-specific PCR (MSP) and sequencing (mapping). Their relationships were analyzed by Fisher's exact test using the software SPSS. RESULTS: The average mRNA level of Dnmt1 gene from cancerous tissue was higher and that of mbd2 gene from cancerous tissue was lower than that from non cancerous tissue, respectively. mbd2 was lower in cancerous tissue than in non cancerous tissue in 14 (50.0%) of patients but higher in 3 cases (10.7%) of non cancerous gastric tissue (P<0.001). c-myc expression was up-regulated in cancer tissues (P<0.05). The up-regulation of mbd2 was found in all patients with hypomethylated c-myc. The transcriptional levels of p16(INK4A) and MeCP2 genes did not display any difference between gastric cancerous and matched non cancerous tissues. There were down-regulation and hypermethylation of hMSH2 in cancer tissues, and the hypermethylation of hMSH2 coexisted with down-regulated transcription. However, the transcription level of the above genes was not associated with biological behaviours of gastric cancers. CONCLUSION: The up regulation of proto-oncogene may be the consequence of epigenetic control of gene expression by demethylase, and mbd2 is involved in the regulation of hMSH2 expression in human gastric cancer. PMID- 15526355 TI - Transabdominal ultrasonography in preoperative staging of gastric cancer. AB - AIM: To investigate the value of transabdominal ultrasonography (US) in the preoperative staging of gastric cancer. METHODS: A total of 198 patients with gastric cancer underwent preoperatively transabdominal US, depth of tumor infiltration was assessed in 125 patients, and lymph node metastasis was assessed in 106 patients. RESULTS: The staging accuracy of transabdominal US was 55.6%, 75.0%, 87.3% and 71.1% in T1, T2, T3 and T4 carcinomas, respectively. The overall accuracy was 77.6%. The detection rate for pancreatic invasion and liver invasion was 77.4%, 71.4%, respectively. The sensitivity, specificity, accuracy of transabdominal US in assessment of lymph node metastasis were 77.6%, 64.1%, 72.6%, respectively. Various shapes such as round, ovoid, spindle were encountered in benign and malignant lymph nodes. Majority of both benign and malignant lymph nodes were hyperechoic and had a distinct border. Benign lymph nodes were smaller than malignant lymph nodes in length and width (P = 0.000, 0.005). Irregular shape, fusional shape, infiltrative signs, inhomogenous echo were seen mainly in malignant lymph nodes (P = 0.045, 0.006, 0.027, 0.006). CONCLUSION: Transabdominal US is useful for preoperative staging in gastric cancer, although it is difficult to differentiate benign from malignant lymph nodes. PMID- 15526356 TI - Surgical treatment and prognosis of gastric cancer in 2,613 patients. AB - AIM: To analyze the factors influencing the prognosis of patients with gastric cancer after surgical treatment, in order to optimize the surgical procedures. METHODS: A retrospective study of 2 613 consecutive patients with gastric cancer was performed. Of these patients, 2,301 (88.1%) received operations; 196 explorative laparotomy (EL), 130 by-pass procedure (BPP), and 1 975 surgical resection of the tumors (891 palliative resection and 1 084 curative resection). The survival rate was calculated by the actuarial life table method, and the prognostic factors were evaluated using the Cox regression proportional hazard model. RESULTS: Of the patients, 2,450 (93.8%) were followed-up. The median survival period was 4.6 mo for patients without operation, 5.2 mo for EL, 6.4 mo for BPP, and 15.2 mo for palliative resection (P = 0.0001). Of the patients with surgical resection of the tumors, the overall 1, 3 and 5-year survival rates after were 82.7%, 46.3% and 31.1%, respectively, with the 5-year survival rate being 51.2% in patients with curative resection, and 7.8% for those with palliative resection. The 5-year survival rate was 32.5% for patients with total gastrectomy, and 28.3% for those with total gastrectomy plus resection of the adjacent organs. The factors that independently correlated with poor survival included advanced stage, upper third location, palliative resection, poor differentiation, type IV of Borrmann classification, tumor metastasis (N3), tumor invasion into the serosa and contiguous structure, proximal subtotal gastrectomy for upper third carcinoma and D1 lymphadenectomy after curative treatment. CONCLUSION: The primary lesion should be resected as long as the local condition permitted for stage III and IV tumors, in order to prolong the patients' survival and improve their quality of life after operation. Total gastrectomy is indicated for carcinomas in the cardia and fundus, and gastric cancer involving the adjacent organs without distant metastasis requires gastrectomy with resection of the involved organs. PMID- 15526357 TI - Epstein-Barr virus in hepatocellular carcinogenesis. AB - AIM: In recent years, studies have suggested that Epstein-Barr virus (EBV) is associated with HCC. The present study was to determine the prevalence of EBV in HCC patients, and whether EBV acted synergistically with hepatitis viruses in HCC carcinogenesis. METHODS: Liver tissue 115 HCC patients and 26 non-carcinoma patients were studied. Polymerase chain reaction (PCR) was performed to detect EBV BamHI W DNA, EBV LMP1 DNA, HBV X DNA, and HBV S DNA. Reverse transcription PCR (RT-PCR) was performed to detect HCV RNA and HDV RNA. Immunohistochemistry was performed to detect LMP1, HBsAg, HBcAg and HCV. The positive ratios were compared between HCC group and control group by chi2 test. RESULTS: Totally, 78 HCC samples whose beta-globulin DNA was positively detected by amplified PCR were selected. PCR was performed in all cases for EBV DNA and HBV DNA. RT-PCR was performed in 18 cases for HCV RNA and HDV RNA. EBV BamHI W and EBV LMP1 were positive in 18 and 6 cases, respectively. HBV X gene and HBV S gene were positive in 42 and 27 cases respectively. HCV was positive in one of the 18 cases, and none was positive for HDV. The positive rates were 28.2% (22 of 78) for EBV DNA (BamHI W and/or LMP1) and 56.4% (44 of 78) for HBV DNA (X gene and/or S gene) respectively. In addition, 12 cases were positive for both EBV DNA and HBV DNA. Among the 26 cases in the control group, 2 cases were positive for EBV BamHI W, 4 positive for HBV X gene and 3 positive for HBV S gene. The positive rates were 8.0% (2 of 26) and 23.1% (6 of 26), respectively, for EBV DNA and HBV DNA. The result of DNA sequencing of BamHI W was 100% homologous with the corresponding sequence of B95-8. There was significant difference in EBV infection rate between HCC patients and controls (chi2 = 4.622, P<0.05). The difference in HBV infection rate was also significant (chi2 = 8.681, P<0.05). However, there was no obvious correlation between HBV and EBV in HCC patients (chi2 = 0.835, P>0.05). LMP1, HBV (HBsAg, HBcAg) and HCV were detected positively in 25, 45 and 6 of 78 cases of HCC tissues respectively. In the 26 control cases, the corresponding positive cases were 2, 4 and 0. The difference in EBV infection rate between HCC patients and control cases was statistically significant (chi2 = 6.02, P<0.05). The difference in HBV infection rate was also statistically significant (chi2 = 10.03, P<0.05). In the 25 cases with positive LMP1 expression, 6 were in the nuclei of tumor cells, 9 in the cytoplasm of tumor cells and 10 in mesenchymal lymphocyte cytoplasm. CONCLUSION: The existence of EBV infection in HCC tissues suggests that EBV may be involved in the hepatocellular carcinogenesis in China. HBV infection may be a major cause of HCC. There is no correlation between EBV and HBV in the development of HCC. The prevalence of HCV infection is low in our area, and HDV appears not to play a direct role in hepatocellular carcinogenesis. PMID- 15526358 TI - Expressions of cysteine-rich61, connective tissue growth factor and Nov genes in hepatocellular carcinoma and their clinical significance. AB - AIM: To investigate the expression of cysteine-rich61 (Cyr61), connective tissue growth factor (CTGF) and nephroblastoma overexpressed gene (Nov) in hepatocellular carcinoma (HCC), and to evaluate the relationship between Cyr61, CTGF and Nov genes expression with invasion and metastasis of HCC. METHODS: Thirty-one HCC specimens were divided into small hepatocellular carcinoma (SHCC), nodular hepatocellular carcinoma (NHCC), solitary large hepatocellular carcinoma (SLHCC) according to their diameter and number of nodes. Reverse transcription polymerse chain reaction (RT-PCR) was used to detect the mRNA expression levels of Cyr61, CTGF and Nov genes in 31 resected specimens of hepatocellular carcinoma and para-cancerous normal liver tissues semi-quantitatively and the relation between their expression levels and clinical pathological parameters were compared. RESULTS: The expressions of Cyr61 and CTGF mRNA in carcinoma tissues were significantly higher than those in para-cancerous normal liver tissues (P<0.01). The expressions of Cyr61 and CTGF mRNA in HCC with venous invasion were higher than those in HCC without venous invasion. CTGF expression in HCC Edmondson's grade III-IV was significantly higher than that in HCC Edmondson's grade I-II (P = 0.022). There was no obvious correlation between Nov mRNA and clinical-pathological features. Compared to NHCC, SLHCC had better cell differentiation, easier capsule formation, less microscopic venous invasion, milder liver cirrhosis. The expressions of Cyr61 and CTGF mRNA in NHCC were significantly higher than those in SLHCC and SHCC. CONCLUSION: Cyr61 and CTGF genes may play an important role in hepatocellular carcinogenesis and correlate with recurrence and metastasis of hepatocellular carcinoma. SLHCC has better biological behaviors than NHCC. PMID- 15526359 TI - Peroxisome proliferator-activated receptor gamma ligands suppress liver carcinogenesis induced by diethylnitrosamine in rats. AB - AIM: Peroxisome proliferator-activated receptor gamma (PPARgamma) is known to regulate growth arrest and terminal differentiation of adipocytes and is used clinically as a new class of antidiabetic drugs. Recently, several studies have reported that treatment of cancer cells with PPARgamma ligands could induce cell differentiation and apoptosis, suggesting a potential application as chemopreventive agents against carcinogenesis. In the present study, 3 different kinds of PPARgamma ligands were subjected to the experiments to confirm their suppressive effects on liver carcinogenesis. METHODS: Three PPARgamma ligands, pioglitazone (Pio) (200 ppm), rosiglitazone (Rosi) (200 ppm), and troglitazone (Tro) (1,000 ppm) were investigated on the induction of the placental form of rat glutathione S-transferase (rGST P) positive foci, a precancerous lesion of the liver, and liver cancer formation using a diethylnitrosamine-induced liver cancer model in Wistar rats, and dose dependency of a PPARgamma ligand was also examined. RESULTS: PPARgamma ligands reduced the formation of rGST P-positive foci by diethylnitrosamine and induction of liver cancers was also markedly suppressed by a continuous feeding of Pio at 200 ppm. CONCLUSION: PPARgamma ligands are potential chemopreventive agents for liver carcinogenesis. PMID- 15526360 TI - Preparation of human hepatocellular carcinoma-targeted liposome microbubbles and their immunological properties. AB - AIM: To prepare the human hepatocellular carcinoma-(HCC)-targeted liposome microbubbles and to investigate their immunological properties. METHODS: Human hepatocarcinoma specific monoclonal antibody HAb18 was attached to the surface of home-made liposome microbubbles by static attraction to prepare the targeted liposome microbubbles. The combination of HAb18 with liposome microbubbles was confirmed by the slide agglutination test and immunofluorescent assay. Their immunological activity was measured by ELISA. Rosette formation test, rosette formation blocking test and immun-ofluorescent assay were used to identify the specific binding of targeted liposome microbubbles to SMMC-7721 hepatoma cells, and cytotoxicity assay was used to detect their effect on human hepatocytes. RESULTS: The targeted liposome microbubbles were positive in the slide agglutination test and immunofluorescent assay. ELISA indicated that the immunological activity of HAb18 on the liposome microbubbles was similar to that of free HAb18. SMMC-7721 cells were surrounded by the targeting liposome microbubbles to form rosettes, while the control SGC-7901 gastric cancer cells were not. Proliferation of SMMC-7721 cells and normal human hepatocytes was not influenced by the targeted liposome microbubbles. CONCLUSION: The targeted liposome microbubbles with a high specific biological activity have been successfully prepared, which specifically bind to human hepatocarcinoma cells, and are non-cytotoxic to hepatocytes. These results indicate that the liposome microbubbles can be used as a HCC-targeted ultrasound contrast agent that may enhance ultrasound images and thus improve the diagnosis of HCC, especially at the early stage. PMID- 15526361 TI - Effect of hepatoma H22 on lymphatic endothelium in vitro. AB - AIM: To determine the effect of metastatic hepatoma cells on lymphangioma-derived endothelium, and to establish in vitro model systems for assessing metastasis related response of lymphatic endothelium. METHODS: Benign lymphangioma, induced by intraperitoneal injection of the incomplete Freund's adjuvant in BALB/c mice, was embedded in fibrin gel or digested and then cultured in the conditioned medium derived from hepatoma H22. Light and electron microscopy, and the transwell migration assay were used to determine the effect of H22 on tissue or cell culture. Expressions of Flt-4, c-Fos, proliferating cell nuclear antigen (PCNA), and inducible nitric oxide synthase (iNOS) in cultured cells, and content of nitric oxide in culture medium were also examined. RESULTS: The embedded lymphangioma pieces gave rise to array of capillaries, while separated cells from lymphangioma grew to a cobblestone-like monolayer. H22 activated growth and migration of the capillaries and cells, induced expressions of Flt-4, c-Fos, PCNA and iNOS in cultured cells, and significantly increased the content of NO in the culture medium. CONCLUSION: Lymphangioma-derived cells keep the differentiated phenotypes of lymphatic endothelium, and the models established in this study are feasible for in vitro study of metastasis-related response of lymphatic endothelium. PMID- 15526362 TI - Methylation profile of the promoter CpG islands of 14 "drug-resistance" genes in hepatocellular carcinoma. AB - AIM: To establish the DNA methylation patterns of the promoter CpG islands of 14 "drug-resistance" genes in hepatocellular carcinoma (HCC). METHODS: The methylation specific polymerase chain reaction in conjunction with sequencing verification was used to establish the methylation patterns of the 14 genes in the liver tissues of four healthy liver donors, as well as tumor and the paired non-cancerous tissues of 30 HCC patients. RESULTS: While 11 genes (ATP-binding cassette, sub-family G (WHITE), member 2(ABCG2), activating transcription factor (ATF2), beta-2-microglobulin (B2M), deoxycytidine kinase (DCK), occludin (OCLN), v-raf-1 murine leukemia viral oncogene homolog (RAF1), ralA binding protein 1 (RALBP1), splicing factor (45 kD) (SPF45), S-phase kinase-associated protein 2 (p45) (SKP2), tumor protein p53 (Li-Fraumeni syndrome) (TP53) and topoisomerase (DNA) II beta (TOP2B)) maintained the unmethylated patterns, three genes displayed to various extents the hypermethylation state in tumor tissues in comparison with the normal counterparts. The catalase (CAT) was hypermethylated in tumor and the neighboring non-cancerous tissue of one case (3.3%). Both glutathione S-transferase pi (GSTpi) (80%, 24/30 in tumor and 56.7%, 17/30 in the paired non-cancerous tissues) and cystic fibrosis transmembrane conductance regulator, ATP-binding cassette (sub-family C, member 7) (CFTR) (77%, 23/30 in tumor and 50%, 15/30 in the paired non-cancerous tissues) genes were prevalently hypermethylated in HCC as well as their neighboring non-cancerous tissues. No significant difference in the hypermethylation occurrence was observed between the HCC and its neighboring non-cancerous tissues. CONCLUSION: Hypermethylation of promoter CpG islands of both CFTR and GSTpi genes occurs prevalently in HCC, which may correlate with the low expression of these two genes at the mRNA level and has the profound etiological and clinical implications. It is likely to be specific to the early phase of HCC carcinogenesis. PMID- 15526363 TI - Methylation profile of the promoter CpG islands of 31 genes that may contribute to colorectal carcinogenesis. AB - AIM: To establish the methylation profile of the promoter CpG islands of 31 genes that might play etiological roles in colon carcinogenesis. METHODS: The methylation specific PCR in conjunction of sequencing verification was used to establish the methylation-profile of the promoter CpG islands of 31 genes in colorectal cancer (n = 65), the neighboring non-cancerous tissues (n = 5), colorectal adenoma (n = 8), and normal mucosa (n = 1). Immunohistochemically, expression of 10 genes was assessed on the home-made tissue microarrays of tissues from 58 patients. The correlation of tumor specific changes with each of clinical-pathologic features was scrutinized with relevant statistic tools. RESULTS: In comparison with the normal mucosa of the non-cancer patients, the following 14 genes displayed no tumor associated changes: breast cancer 1, early onset (BRCA1), cadherin 1, type 1, E-cadherin (epithelial) (CDH1), death associated protein kinase 1 (DAPK1), DNA (cytosine-5-)-methyltransferase 1 (DNMT1), melanoma antigen, family A, 1 (directs expression of antigen MZ2-E) (MAGEA1), tumor suppressor candidate 3 (N33), cyclin-dependent kinase inhibitor 1A (p21, Cip1) (p21(WAF1)), cyclin-dependent kinase inhibitor 1B (p27, Kip1) (p27(KIP1)), phosphatase and tensin homolog (mutated in multiple advanced cancers 1) (PTEN), retinoic acid receptor, beta (RAR- , Ras association (RalGDS/AF-6) domain family 1 C (RASSF1C), secreted frizzled-related protein 1 (SFRP1), tissue inhibitor of metalloproteinase 3 (Sorsby fundus dystrophy, pseudoinflammatory) (TIMP3), and von Hippel-Lindau syndrome (VHL). The rest 17 targets exhibited to various extents the tumor associated changes. As changes in methylation of the following genes occurred marginally, their impact on the formation of colorectal cancer were trivial: adenomatous polyposis coli (APC) (8%, 5/65), Ras association (RalGDS/AF-6) domain family 1A (RASSF1A) (3%, 2/65) and cyclin-dependent kinase inhibitor 2A, alternated reading frame (p14(ARF)) (6%, 4/65). The following genes exhibited moderate changes in methylation: O-6-methylguanine-DNA methyltransferase (MGMT) (20%, 13/65), mutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) (hMLH1) (18%, 12/65), cyclin-dependent kinase inhibitor 2A (melanoma, p16, inhibits CDK4) (p16(INK4a)) (10%, 10/65), methylated in tumor 1 (MINT1) (15%, 10/65), methylated in tumor 31 (MINT31) (11%, 7/65). The rest changed greatly in the methylation pattern in colorectal cancer (CRC): cyclin A1 (cyclin a1) (100%, 65/65), caudal type homeobox transcription factor 1 (CDX1) (100%, 65/65), RAR- (85%, 55/65), myogenic factor 3 (MYOD1) (69%, 45/65), cyclin dependent kinase inhibitor 2B (p15, inhibits CDK4) (p15(INK4b)) (68%, 44/65), prostaglandin-endoperoxide synthase 2 (prostaglandin G/H synthase and cyclooxygenase) (COX2) (72%, 47/65), cadherin 13, H-cadherin (heart) (CDH13) (65%, 42/65), CAAX box 1 (CXX1) (58%, 38/65), tumor protein p73 (p73) (63%, 41/65) and Wilms tumor 1 (WT1) (58%, 38/65). However, no significant correlation of changes in methylation with any given clinical-pathological features was detected. Furthermore, the frequent changes in methylation appeared to be an early phase event of colon carcinogenesis. The in situ expression of 10 genes was assessed by the immunohistochemical approach at the protein level: CDH1, CDH13, COX2, cyclin A1, hMLH1, MGMT, p14(ARF), p73, RAR- , and TIMP3 genes in the context of the methylation status in colorectal cancer. No clear correlation between the hypermethylation of the promoter CpG islands and the negative expression of the genes was established. CONCLUSION: The methylation profile of 31 genes was established in patients with colon cancer and colorectal adenomas, which provides new insights into the DNA methylation mediated mechanisms underlying the carcinogenesis of colorectal cancer and may be of prognostic values for colorectal cancer. PMID- 15526364 TI - Effects of phosphorothioate anti-sense oligodeoxynucleotides on colorectal cancer cell growth and telomerase activity. AB - AIM: To investigate the inhibitory effect of phosphorothioate anti-sense oligodeoxynucleotides (PASODN) on colorectal cancer LS-174T cells in vitro and the mechanism of inhibition of telomerase activity in these cells. METHODS: PASODN were used to infect LS-174T cells and block human telomerase RNA (hTR) through anti-sense technology. The inhibitory effect of PASODN was evaluated by colony-forming inhibition assay and growth curve. Changes of telomerase activity in LS-174T cells were detected by polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA), and the level of apoptosis was analyzed by flow cytometry (FCM) assay. RESULTS: PASODN showed a dose and time-dependent inhibition of cell proliferation. The optimal dosage of PASODN was 10 mumol/L. The colony-forming efficiency was 10.3% in PASODN group after 10 d, whereas that in phosphorothioate mis-sense oligodeoxynucleotides (PMSODN) group with the same concentration and in PBS group (blank control) was 49.1% and 50.7%, respectively. PCR-ELISA results indicated that telomerase activity in the PASODN group was obviously inhibited in comparison with in the control groups (P<0.01, t = 3.317 and 3.241, t0.01(20) = 2.845). Meanwhile, before the number of cells was decreased, the morphological changes were observed in the cells of PASODN group. The cells in PASODN group showed the apoptotic peak at 72 h after infection, whereas the control group did not show. CONCLUSION: Specific sequence oligonucleotides can inhibit telomerase activity and lead to cell apoptosis, suggesting a novel treatment strategy for malignant tumors induced by telomerase. PMID- 15526365 TI - Honokiol: a potent chemotherapy candidate for human colorectal carcinoma. AB - AIM: To investigate the anticancer activity of honokiol on RKO, a human colorectal carcinoma cell line in vitro and in vivo, and to evaluate its possible use in clinic. METHODS: In vitro anticancer activity of honokiol was demonstrated by its induction of apoptosis in tumor cells. We analyzed cell proliferation with MTT assay, cell cycle with flow cytosmeter, DNA fragment with electrophoresis on agarose gels. To test the mechanism of honokiol-induced apoptosis, Western blotting was used to investigate the factors involved in this process. The pharmacokinetics study of honokiol was tested by high phase liquid chromatography. In in vivo study, Balb/c nude mice were incubated with RKO cells. Honokiol was injected intraperitoneally every other day into tumor bearing Balb/c nude mice. RESULTS: Our results showed that honokiol induced apoptosis of RKO cells in a time- and dose-dependent manner. At 5-10 microg/mL for 48 h, honokiol induced apoptosis through activating Caspase cascades. Pharmacokinetics study demonstrated that, honokiol could be absorbed quickly by intraperitoneal injection, and maintained in plasma for more than 10 h. In nude mice bearing RKO incubated tumor, honokiol displayed anticancer activity by inhibiting tumor growth and prolonging the lifespan of tumor bearing mice. CONCLUSION: With its few toxicity to normal cells and potent anticancer activity in vitro and in vivo, honokiol might be a potential chemotherapy candidate in treating human colorectal carcinoma. PMID- 15526366 TI - Diagnosis of Helicobacter pylori infection and diseases associated with Helicobacter pylori by Helicobacter pylori outer membrane proteins. AB - AIM: To examine the serological response of patients with upper gastrointestinal diseases and Helicobocter pylori (H pylori) infection to two H pylori outer membrane proteins (OMPs) (Mr18,000 and Mr 26,000) acquired by gene recombinant technique, and to determine the diagnostic significance of serological tests derived from these OMPs. METHODS: Recombinant vectors encoding the two H pylori OMPs were used to transform and express in BL21 (DE3) E.coli. After purification with Ni2+-NTA agarose resin, colloid gold kits were prepared with purified recombinant proteins to detect H pylori infection and H pylori-associated diseases by the immunity-marker technology. We selected 150 patients with H pylori infection and digestive symptoms without previous treatment, including chronic gastritis (n = 60), duodenal ulcer (n = 30), gastric ulcer (n = 30), and gastric cancer (n = 30). As controls, 33 H pylori-negative healthy volunteers were also recruited. Serum samples were collected from all subjects, and the antibodies to specific proteins of H pylori were tested with the colloid gold test kits. The sensitivity, specificity and accuracy of the colloid gold tests were evaluated, by using the combination of standard diagnostic methods (13C urea breath test and bacteria culture) and classic enzyme-linked immunosorbent assay (ELISA) as reference. RESULTS: After purification with Ni2+-NTA agarose resin, the purity of recombinant fusion proteins was about 95%. The recombinant fusion proteins were recognized by the specific monoclonal antibodies against the two H pylori OMPs, as demonstrated by the ELISA. Of the 150 serum samples from patients infected with H pylori 141 (94.0%) responded positively to the recombinant protein with Mr 26,000, while the seropositive rates were 95.0%, 96.7%, 96.7% and 90.0% for patients with H pylori-associated chronic gastritis, duodenal ulcer, gastric ulcer, and gastric cancer respectively. The sensitivity, specificity, and accuracy of the colloid gold kit with Mr 26,000 protein were 94.0%, 97.0%, and 94.5%, respectively. Compared with the classic ELISA, bacteria culture and 13C urea breath test results in detecting H pylori-infection, there was no significant difference (P>0.05). For the colloid gold kit with Mr 18,000, the seropositive rates were 52.0%, 40.0%, 40.0%, 53.3% and 86.7%, respectively, in H pylori-infected patients, and those with H pylori-associated chronic gastritis, duodenal ulcer, gastric ulcer, and gastric cancer. There was a significant difference (P<0.05) in seropositivity between patient with gastric cancer (86.7%) and those with other diseases (43.3%). CONCLUSION: The two colloid gold kits derived from the recombinant OMPs are useful tools either for detecting H pylori infection, or for, predicting H pylori-associated gastric malignancy. PMID- 15526367 TI - Ethanol inhibits the motility of rabbit sphincter of Oddi in vitro. AB - AIM: The role of the sphincter of Oddi (SO) in ethanol (ETOH)-induced pancreatitis is controversial. Our aim was to characterise the effect of ETOH on basal and stimulated SO motility. METHODS: SOs removed from white rabbits were placed in an organ bath (Krebs solution, pH7.4, 37 degrees). The effects of 2 mL/L, 4 mL/L, 6 mL/L and 8 mL/L of ETOH on the contractile responses of the sphincter were determined. SOs were stimulated with either 0.1 mumol/L carbachol, 1 mumol/L erythromycin or 0.1 mumol/L cholecystokinin (CCK). RESULTS: ETOH at a dose of 4 mL/L significantly decreased the baseline contractile amplitude from 11.98+/-0.05 mN to 11.19+/-0.07 mN. However, no significant changes in the contractile frequency were observed. ETOH (0.6%) significantly decreased both the baseline amplitude and the frequency compared to the control group (10.50+/-0.01 mN, 12.13+/-0.10 mN and 3.53+/-0.13 c/min, 5.5+/-0.13 cycles(c)/min, respectively). Moreover, 0.8% of ETOH resulted in complete relaxation of the SO. Carbachol (0.1 micromol/L) or erythromycin (1 micromol/L) stimulated the baseline amplitudes (by 82% and 75%, respectively) and the contractile frequencies (by 150% and 106%, respectively). In the carbachol or erythromycin-stimulated groups 2-6 mL/L of ETOH significantly inhibited both the amplitude and the frequency. Interestingly, a 4-5 min administration of 0.6% ETOH suddenly and completely relaxed the SO. CCK (0.1 micromol/L) stimulated the baseline amplitude from 12.37+/-0.05 mN to 27.40+/-1.82 mN within 1.60+/-0.24 min. After this peak, the amplitude decreased to 17.17+/-0.22 mN and remained constant during the experiment. The frequency peaked at 12.8+/-0.2 c/min, after which the constant frequency was 9.43+/-0.24 c/min throughout the rest of the experiment. ETOH at a dose of 4 mL/L significantly decreased the amplitude from 16.13+/-0.23 mN to 14.93+/-0.19 mN. However, no significant changes in the contractile frequency were observed. ETOH at a dose of 6 mL/L inhibited both the amplitudes and the frequencies in the CCK-stimulated group, while 8 mL/L of ETOH completely relaxed the SO. CONCLUSION: ETOH strongly inhibits the basal, carbachol, erythromycin, and CCK-stimulated rabbit SO motility. Therefore, it is possible that during alcohol-intake the relaxed SO opens the way for pancreatic fluid to flow out into the duodenum in rabbits. This relaxation of the SO may protect the pancreas against alcohol-induced damage. PMID- 15526368 TI - DNA ploidy and c-Kit mutation in gastrointestinal stromal tumors. AB - AIM: To investigate the prognostic significance of c-Kit gene mutation and DNA ploidy in gastrointestinal stromal tumors (GISTs). METHODS: A total of 55 cases of GISTs were studied for the expression of c-Kit by immunohistochemistry, and the c-Kit gene mutations in exons 9, 11, 13, and 17 were detected by polymerase chain reaction-single strand confirmation polymarphism (PCR-SSCP) and denaturing high performance liquid chromatography (D-HPLC) techniques. DNA ploidy was determined by flow cytometry. RESULTS: Of the 55 cases of GISTs, 53 cases (96.4%) expressed c-Kit protein. The c-Kit gene mutations of exons 11 and 9 were found in 30 (54.5%) and 7 cases (12.7%), respectively. No mutations were found in exons 13 and 17. DNA aneuploidy was seen in 10 cases (18.2%). The c-Kit mutation positive GISTs were larger in size than the negative GISTs. The aneuploidy tumors were statistically associated with large size, high mitotic counts, high risk groups, high cellularity and severe nuclear atypia, and epithelioid type. There was a tendency that c-Kit mutations were more frequently found in aneuploidy GISTs. CONCLUSION: DNA aneuploidy and c-Kit mutations can be considered as prognostic factors in GISTs. PMID- 15526369 TI - Genomic determination of CR1 CD35 density polymorphism on erythrocytes of patients with gallbladder carcinoma. AB - AIM: To study the changes of quantitative expression, adhering activity and genomic density polymorphism of complement types in erythrocytes (CR1) of patients with gallbladder carcinoma and the related clinical significance. METHODS: Polymerase chain reaction (PCR), Hind III restriction enzyme digestion, quantitative assay of CR1 and adhering activity assay of CR1 in erythrocytes were used. RESULTS: The number and adhering activity of CR1 in patients with gallbladder carcinoma (0.738+/-0.23, 45.9+/-5.7) were significantly lower than those in chronic cholecystitis and cholecystolithiasis (1.078+/-0.21, 55.1+/-5.9) and healthy controls (1.252+/-0.31, 64.2+/-7.4) (P<0.01). The number and adhering activity of CR1 in patients with chronic cholecystitis and cholecystolithiasis (1.078+/-0.21, 55.1+/-5.9) were significantly lower than those in healthy controls (1.252+/-0.31, 64.2+/-7.4) (P<0.05). There was a positive correlation between quantitative expression and adhering activity of CR1 (r = 0.79, P<0.01). Compared with those on preoperative day (0.738+/-0.23, 45.4+/-4.9), the number and adhering activity of CR1 in patients with gallbladder carcinoma decreased greatly on the third postoperative day (0.310+/-0.25, 31.8+/-5.1) (P<0.01), and on the first postoperative week (0.480+/-0.25, 38.9+/-5.2) (P<0.01), but they were increased slightly than those on the preoperative day (P>0.05). The number and adhering activity of CR1 recovered in the second postoperative week(0.740+/ 0.24, 46.8+/-5.9) (P<0.01) and increased greatly in the third postoperative week (0.858+/-0.35, 52.7+/-5.8) (P<0.01) in comparison with those on the preoperative day and in the first postoperative week. The number and adhering activity of CR1 of gallbladder carcinoma patients with infiltrating, adjacent lymphogenous and distant organ metastases were significantly lower than those of gallbladder carcinoma patients without them (P<0.01). No difference was observed between the patients with gallbladder carcinoma and healthy individuals in the spot mutation rate of CR1 density gene (chi(2) = 0.521, P>0.05). The distribution of expression was 67.8% in high expression genomic type, 24.8% in moderate expression genomic type, and 7.4% in low expression genomic type. The number and adhering activity of CR1 high expression genomic type gallbladder carcinomas (0.749+/-0.22, 42.1+/ 6.2) were significantly lower than those of healthy individuals (1.240+/-0.29, 63.9+/-7.2), and were also significantly lower than those of healthy individuals (0.921+/-0.23, 54.8+/-7.1), but no difference was observed between the number and adhering activity of CR1 lower expression genomic type gallbladder carcinomas (0.582+/-0.18, 44.3+/-5.5) and those of healthy individuals (0.610+/-0.20, 45.8+/ 5.7) (P>0.05). CONCLUSION: Defective expression of CR1 in gallbladder carcinoma is mostly acquired through central peripheral mechanisms. The changes in CR1 quantitative expression and adhering activity are consanguineously related to the development and metastasis in gallbladder carcinoma. PMID- 15526370 TI - Effects of dermatan sulfate derivatives on platelet surface P-selectin expression and protein C activity in blood of inflammatory bowel disease patients. AB - AIM: To investigate the effect of dermatan sulfate (DS) derivatives on platelet surface P-selectin expression and blood activated protein C (APC) activity in patients with inflammatory bowel disease (IBD), and to clarity the anti inflammatory mechanism of DS derivatives. METHODS: Dermatan sulfate (DS) was sulfated with chlorosulfonic acid to prepare polysulfated dermatan sulfate (PSDS). The major disaccharides of DS and PSDS were determined by 1H nuclear magnetic resonance spectroscopy (1H-NMR) and 13C-NMR. Both DS and PSDS were depolymerized with hydrogen peroxide. The fragments were separated by gel filtration chromatography. The effects of DS derivatives on P-selectin expression were assayed by ELISA method, and blood APC activity was assayed by the synthetic chromogenic substrate method. RESULTS: The major disaccharides of DS and PSDS were IdoA-1-3-GalNAc-4-SO3 and IdoA-2SO3-1-3-GalNAc4, 6-diSO3, respectively. Compared with the adenosine diphosphate stimulated group and IBD control group, DS and its derivatives all had significant inhibitory effects on P-selectin expression (P<0.01), but there was no difference between DS-derived oligosaccharides (DSOSs) and PSDS-derived oligosaccharides (PSDSOSs). The experiments on APC activity showed that DS and its derivatives all enhanced APC activity. The most active DSOS was the one with a relative molecular weight (Mr) of 4,825, which enhanced the APC activity from 106.5+/-11.5% to 181.8+/-22.3% (P<0.01). With the decrease of Mr, the activity of DSOSs decreased gradually. The effect of PSDS on APC activity enhancement was more significant than that of DS, and the APC activity was raised to 205.2+/-22.1% (P<0.01). All the PSDSOSs were more active than DSOSs on the basis of comparable Mr. With the decrease of Mr, the activity of PSDSOSs increased gradually, and the most active PSDSOS was PSDSOS3 with Mr of 2,749, which enhanced the APC activity to 331.2+/-27.8% (P<0.01), then the activity of PSDSOSs decreased gradually. CONCLUSION: DS and its derivatives can significantly inhibit P-selectin expression on platelet surface, but the effect has no correlation with DS molecular mass and sulfation. The effect of DS or its derivatives on APC activity at molecular level involves complex mechanisms that depend on the molecular mass, the degree of sulfation, and the heterogeneous composition of DS. On the same molecular size, the higher the degree of DS sulfation, the more significant the effect on enhancing APC activity. PMID- 15526371 TI - Inhibitory effect of heparin-derived oligosaccharides on secretion of interleukin 4 and interleukin-5 from human peripheral blood T lymphocytes. AB - AIM: To investigate the inhibitory effect of heparin-derived oligosaccharides (Oligs) on secretion of interleukin-4 (IL-4) and interleukin-5 (IL-5) from human peripheral blood T lymphocytes (PBTLs). METHODS: Oligs were prepared by three different heparin depolymerization methods and separated by gel filtration chromatography. PBTLs from ten adult patients with allergic eosinophilic gastroenteritis were treated with phytahematoagglutinin (PHA) and Oligs. The supernatants from the cell culture of PBTLs were harvested and subjected to the determination of IL-4 and IL-5 contents by ELISA method. RESULTS: At the concentration of 5 microg/mL, Oligs with different Mr had different effects on the secretion of IL-4 and IL-5. The tetrasaccharide with Mr of 1,142, produced by depolymerizing heparin with hydrogen peroxide, had the strongest inhibitory effect on the secretion of IL-4. It decreased the IL-4 content from 375.6+/-39.2 ng/L (PHA group) to 12.5+/-5.7 ng/L (P<0.01). The hexasaccharide with Mr of 1,806, produced by depolymerizing heparin with beta-elimination method, had the strongest inhibitory effect on the secretion of IL-5. It decreased the IL-5 content from 289.2+/-33.4 ng/L (PHA group) to 22.0+/-5.2 ng/L (P<0.01). CONCLUSION: The inhibitory activity of Oligs on the secretion of IL-4 and IL-5 from human PBTLs closely depends on their molecular structure, and there may be an essential structure to act as an inhibitor. The most effective inhibitors of IL-4 and IL-5 secretion are tetrasaccharides and hexasaccharides, respectively. PMID- 15526372 TI - Selection criteria for preoperative endoscopic retrograde cholangiopancreatography before laparoscopic cholecystectomy and endoscopic treatment of bile duct stones: results of a retrospective, single center study between 1996-2002. AB - AIM: The optimal treatment for bile duct stones (in terms of cost, complications and accuracy) is unclear. The aim of our study was to determine the predictive factors for preoperative endoscopic retrograde cholangiopancreatography (ERCP). METHODS: Patients undergoing preoperative ERCP (< or =90 d before laparoscopic cholecystectomy) were evaluated in this retrospective study from the 1st of January 1996 to the 31st of December 2002. The indications for ERCP were elevated serum bilirubin, elevated liver function tests (LFT), dilated bile duct (> or =8 mm) and/or stone at US examination, coexisting acute pancreatitis and/or acute pancreatitis or jaundice in patient's history. Suspected prognostic factors and the combination of factors were compared to the result of ERCP. RESULTS: Two hundred and six preoperative ERCPs were performed during the observed period. The rate of successful cannulation for ERC was (97.1%). Bile duct stones were detected in 81 patients (39.3%), and successfully removed in 79 (97.5%). The number of prognostic factors correlated with the presence of bile duct stones. The positive predictive value for one prognostic factor was 1.2%, for two 43%, for three 72.5%, for four or more 91.4%. CONCLUSION: Based on our data preoperative ERCP is highly recommended in patients with three or more positive factors (high risk patients). In contrast, ERCP is not indicated in patients with zero or one factor (low risk patients). Preoperative ERCP should be offered to patients with two positive factors (moderate risk patients), however the practice should also be based on the local conditions (e.g. skill of the endoscopist, other diagnostic tools). PMID- 15526373 TI - Effect of integrated traditional Chinese and Western medicine on SARS: a review of clinical evidence. AB - AIM: To assess the possible effect of integrated traditional Chinese and Western medicine on severe acute respiratory syndromes. METHODS: The current available randomized controlled trials of integrated traditional Chinese and Western medicine on SARS were identified through systematically searching literature in any languages or any types of publications. Additional studies of gray literature were also collected. The quality of studies was evaluated by two investigators independently based largely on the quality criteria specified CONSORT. Statistical analysis of the results was performed using RevMan 4.2.0 software developed by the Cochrane Collaboration. RESULTS: Six studies (n = 366) fulfilling the inclusion criteria were found, of which the quality of one study was graded as B, the remaining five were graded as C. Two studies were performed with meta-analysis, the other four studies existed some heterogeneity for which meta-analysis could not be performed, a significant effect on lung infiltrate absorption was found in the treatment groups of these two studies (RR 6.68, 95% CI (2.93, 15.24), P<0.01), there was no significant differences between the mortality (RR 0.86, 95% CI (0.22, 3.29), P = 0.82) and the average dosage of corticosteroid (WMD -39.65, 95% CI (-116.84, 37.54), P = 0.31). The other three studies also showed significant differences in infiltrate absorption, including national drug No. 2. 3. 4 in combination with Western medicine (RR 5.45, 95% CI (1.54, 19.26)), compound formulas NO. 1 combined with Western medicine (WMD 0.24, 95% CI (0.02, 0.46)), compound formulas combined with Western medicine (RR 8.06, 95% CI (0.40, 163.21)). Kangfeidian No.4 in combination with Western medicine had no significant effect on symptom improvement such as loss of dyspnea and cough (RR 1.50, 95%CI (0.41, 5.43)) and (RR 1.29, 95%CI (0.30, 5.43)). CONCLUSION: Integrated traditional Chinese and Western medicines has some positive effects on lung infiltrate absorption in SARS patients, and is recommended as an adjunct treatment for SARS. However, its effect on SARS requires further careful study due to limited available randomized control trials. PMID- 15526374 TI - HDR-192Ir intraluminal brachytherapy in treatment of malignant obstructive jaundice. AB - AIM: To determine the feasibility and safety of intraluminal brachytherapy in treatment of malignant obstructive jaundice (MOJ) and to evaluate the clinical effect of intraluminal brachytherapy on stent patency and patient survival. METHODS: Thirty-four patients with MOJ were included in this study. Having biliary stent placed, all patients were classified into intraluminal brachytherapy group (group A, n = 14) and control group (group B, n = 20) according to their own choice. Intraluminal brachytherapy regimen included: HDR 192Ir was used in the therapy, fractional doses of 4-7 Gy were given every 3-6 d for 3-4 times, and standard points were established at 0.5-1.0 cm. Some patients of both groups received transcatheter arterial chemoembolization (TACE) after stent placement. RESULTS: In group A, the success rate of intraluminal brachytherapy was 98.0%, RTOG grade 1 acute radiation morbidity occurred in 3 patients, RTOG/EORTC grade 1 late radiation morbidity occurred in 1 patient. Mean stent patency of group A (12.6 mo) was significantly longer than that of group B (8.3 mo) (P<0.05). There was no significant difference in the mean survival (9.4 mo vs 6.0 mo) between the two groups. CONCLUSION: HDR-192Ir intraluminal brachytherapy is a safe palliative therapy in treating MOJ, and it may prolong stent patency and has the potentiality of extending survival of patients with MOJ. PMID- 15526375 TI - Cloning and sequencing of cagA gene fragment of Helicobacter pylori with coccoid form. AB - AIM: To clone and sequence the cagA gene fragment of Helicobacter pylori (H pylori) with coccoid form. METHODS: H pylori strain NCTC11637 were transformed to coccoid form by exposure to antibiotics in subinhibitory concentrations. The coccoid H pylori was collected. cagA gene of the coccoid H pylori strain was amplified by PCR. After purified, the target fragment was cloned into plasmid pMD 18T. The recombinant plasmid pMD-18T-cagA was transformed into E.coli JM109. Positive clones were screened and identified by PCR and digestion with restriction endonucleases. The sequence of inserted fragment was then analysed. RESULTS: cagA gene of 3,444 bp was obtained from the coccoid H pylori genome DNA. The recombinant plasmid pMD-18T-cagA was constructed, then it was digested by BamH I+Sac I, and the product of digestion was identical with the predicted one. Sequence analysis showed that the homology of coccoid and the reported original sequence H pylori was 99.7%. CONCLUSION: The recombinant plasmid containing cagA gene from coccoid H pylori has been constructed successfully. The coccoid H pylori contain completed cagA gene, which may be related to pathogenicity of them. PMID- 15526376 TI - Real-time determination of human telomerase reverse transcriptase mRNA in gastric cancer. AB - AIM: To set up a real-time fluorescent quantitative reverse transcription polymerase chain reaction (RT-PCR) assay, to detect human telomerase reverse transcriptase (hTERT) messenger RNA in gastric carcinomas, and to evaluate quantitative determination of hTERT mRNA in the diagnostic value of gastric carcinomas, and to analyze the correlation between the expression level of hTERT mRNA and clinicopathological parameters in patients with gastric cancer. METHODS: A real-time quantitative RT-PCR (RQ-PCR) based on TaqMan fluorescence methodology and the LightCycler system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and corresponding adjacent non cancerous tissues. The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as internal control and expressed as 100X (hTERT/GAPDH) ratio. Variables were analyzed by the Student's t-test, chi2 test and Fisher's exact test. RESULTS: NhTERT from gastric carcinomas and corresponding adjacent non-cancerous tissues was 6.27+/-0.89 and 0.93+/-0.18, respectively (t = 12.76, P<0.001). There was no significant association between gastric cancer hTERT mRNA expression level and patient's age, gender, tumor size, location and stage (pTNM), but a significant correlation was found between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation. CONCLUSION: Quantitative determination of hTERT mRNA by RQ-PCR is a rapid and sensitive method. hTERT might be a potential biomarker for the early detection of gastric cancer. PMID- 15526377 TI - NDRG2 expression and mutation in human liver and pancreatic cancers. AB - AIM: To investigate the expression of NDRG2 and mutation of the entire coding region of NDRG2 in human liver and pancreatic cancers, and to further discuss the possible causes of NDRG2 distinct expression patterns. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyze the expression of NDRG2 mRNA in 37 fresh cancer specimens (including 8 cases of pancreatic cancer and 29 cases of liver cancer) and adjacent normal tissues collected from clinical operation. In addition, mutation analysis of the whole coding region of NDRG2 in these cancers was examined by polymerase chain reaction single strand conformational polymorphism (PCR-SSCP). RESULTS: Compared with adjacent normal tissues, the expression levels of NDRG2 mRNA in corresponding cancer tissues reduced significantly (pancreatic cancer: 0.680+/-0.112 vs 2.089+/ 0.214, P<0.01) (liver cancer: 0.894+/-0.098 vs 1.345+/-0.177, P<0.05). Using PCR SSCP, the mutation of the whole coding region of NDRG2 was not found in those cancer tissues where the expression of NDRG2 mRNA reduced markedly. CONCLUSION: NDRG2 gene might express differently between normal tissues and cancer tissues, and might play an important role in the development of pancreatic cancer and liver cancer. Low expression of NDRG2 might be unrelated to the mutation of coding region of NDRG2. PMID- 15526378 TI - Involvement of extracellular signal-regulated kinase/mitogen-activated protein kinase pathway in multidrug resistance induced by HBx in hepatoma cell line. AB - AIM: To investigate the molecular mechanism of the influence of HBx protein on multidrug resistance associated genes: multidrug resistance 1 (MDR-1), multidrug related protein (MRP-1), lung resistance related protein (LRP) in hepatoma cells and the potential role of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway in this process. METHODS: A cell model stably expressing the HBx protein was established by liposome-mediated transfection of HBx gene into HepG2 cell line. The expression of multidrug resistance associated genes and proteins was detected by RT-PCR and Western blot. AnnexinV-FITC/PI assay was used to confirm the multidrug resistance (MDR) phenotype of transfected cells by fluorescence cytometry (FACS). The ERK/MAPK pathway activation was measured by Western blot through comparing the ratio of phosphorylation of ERK/MAPK to total ERK/MAPK protein. After treated with the ERK/MAPK pathway inhibitor U0126, the HBx-expressing cells were harvested. Then RT-PCR, Western blot and FACS were used to analyze the alterations in the expression of multidrug resistance associated genes and the MDR phenotype after exposure. RESULTS: Compared with the control group, the transfected cells showed a higher expression of MDR associated genes and proteins. Marked elevations in MDR-1 (64.3%), MRP-1 (87.5%) and LRP (90.8%) were observed in the transfected cells (P<0.05). RT-PCR revealed that the over-expression of MDR associated proteins was due to amplification of such genes (MDR1 2.9 fold, MRP1 1.67 fold, LRP1.95 fold). Furthermore, we found that the ERK/MAPK activity was remarkably high in the HBx expressing cells. The activation of ERK/MAPK, as measured by the ratio of phosphorylated ERK bands normalized to the total ERK bands, was increased by 2.3 fold in HBx-transfected cells compared with cells transfected with the empty vector. After treated with the ERK/MAPK pathway inhibitor, the level of MDR associated genes and proteins in the transfected cells decreased to some extent. Compared with controls, a significant decrease in MDR-1 mRNA (53.3%), MRP-1 mRNA (59.7%) as well as LRP mRNA (56.4%) was observed in the U0126 treated transfected cells after 12 h. Western blot also demonstrated that the protein expression of these MDR associated genes slightly reduced after treated with U0126 for 12 h (MDR-1 40.1%, MRP-1 29.4%, LRP35.7%). This change was accompanied with the rise of cell apoptosis ratio confirmed by Annexin V-PI detection. The apoptosis index of U0126-treated cells increased by 1.28 fold, compared with that of transfected cells. Obviously, the MDR phenotype of these cells was obviously related with increased activities of the ERK/MAPK pathway. CONCLUSION: HBx protein might be one of the causes for the occurrence of MDR in HCC, and ERK/MAPK pathway might be involved in this change. PMID- 15526379 TI - Effect of environmental hyperthermia on gastrin, somatostatin and motilin in rat ulcerated antral mucosa. AB - AIM: To study the effect of environmental hyperthermia on gastrin, somatostatin and motilin in rat ulcerated antral mucosa. METHODS: Forty-two Wistar rats were equally divided into six groups, according to the room temperature (high and normal) and the treatment (acetic acid, normal saline and no treatment). Levels of gastrin, somatostatin and motilin in rat ulcerated antral mucosa were measured with a radioimmunoassay method. RESULTS: The average temperature and humidity were 32.5 degrees and 66.7% for the high temperature group, and 21.1 degrees and 49.3% for the normal temperature group, respectively. Gastric ulcer model was successfully induced in rat injected with 0.05 mL acetic acid into the antrum. In rats with gastric ulcers, the levels of gastrin and motilin increased, whereas the somatostatin level declined in antral mucosa, compared with those in rats treated with normal saline and the controls. However, the change extent in the levels of gastrin, motilin and somatostatin in antral mucosa was less in the high temperature group than in the normal temperature group. CONCLUSION: The levels of gastrin, somatostatin and motilin in rat ulcerated antral mucosal tissue remain relatively stable in a high temperature environment, which may relate to the equilibration of the dynamic system. PMID- 15526380 TI - Effects of selenium on peripheral blood mononuclear cell membrane fluidity, interleukin-2 production and interleukin-2 receptor expression in patients with chronic hepatitis. AB - AIM: To study the effect of selenium on peripheral blood mononuclear cell (PBMC) membrane fluidity and immune function in patients with chronic hepatitis. METHODS: PBMCs were pretreated with selenium (1.156x10(-7) mol/L) for 6 h in vitro or extracted directly from patients after administration of selenium-yeast continuously for 8-12 wk (200 microg/d), and then exposed to Con-A for 48 h. The membrane fluidity, interleukin-2 (IL-2) production and interleukin-2 receptor (IL 2R) expression in PBMCs and malondialdehyde (MDA) concentration in medium and lipid peroxide (LPO) in plasma were determined. RESULTS: The PBMC membrane fluidity, IL-2 production and IL-2R expression in patients with chronic hepatitis were significantly lower than those in healthy blood donators (particle adhesive degree R, 0.17+/-0.01 vs 0.14+/-0.01, P<0.01; IL-2, 40.26+/-9.55 vs 72.96+/ 11.36, P<0.01; IL-2R, 31.05+/-5.09 vs 60.58+/-10.56, P<0.01), and the MDA concentration in medium in patients with chronic hepatitis was significantly higher than that in healthy blood donators (1.44+/-0.08 vs 0.93+/-0.08, P<0.01). Both in vitro and in vivo administration of selenium could reverse the above parameters. CONCLUSION: Supplement of selenium can suppress lipid peroxidation, and improve PBMC membrane fluidity and immune function in patients with chronic hepatitis. PMID- 15526381 TI - Successful management of a benign anastomotic colonic stricture with self expanding metallic stents: a case report. AB - AIM: To assess the effectiveness of and complications associated with metallic stent placement for treatment of anastomotic colonic strictures. METHODS: A 46 year-old man undergoing two procedures of surgery for perforation of descending colon due to a traffic accident presented with pain, abdominal distention, and inability to defecate. Single-contrast barium enema radiograph showed a severe stenosis in the region of surgical anastomosis and the patient was too weak to accept another laparotomy. Under fluoroscopic and endoscopic guidance, we placed two metallic stents in the stenosis site of the anastomosis of the patient with anastomotic colonic strictures. RESULTS: In this case of postsurgical stenosis, the first stent relieved the symptoms of obstruction, but stent migration happened on the next day so an additional stent was required to deal with the stricture and relieve the symptoms. CONCLUSION: This case confirms that metallic stenting may represent an effective treatment for anastomotic colonic strictures in the absence of other therapeutic alternatives. PMID- 15526383 TI - [Radiological quiz of the month: costal lump]. PMID- 15526384 TI - Serum butyrylcholinesterase activity in leprosy. PMID- 15526382 TI - Cyclooxygenase 2, pS2, inducible nitric oxide synthase and transforming growth factor alpha in gastric adaptation to stress. AB - AIM: To determine the role of mucosal gene expression of cyclooxygenase 2 (COX 2), pS2 (belongs to trefoil peptides), inducible nitric oxide synthase (iNOS) and transforming growth factor alpha (TGFalpha) in gastric adaptation to water immersion and restraint stress (WRS) in rats. METHODS: Wistar rats were exposed to single or repeated WRS for 4 h every other day for up to 6 d. Gastric mucosal blood flow (GMBF) was measured by laser Doppler flowmeter-3. The extent of gastric mucosal lesions were evaluated grossly and histologically and expressions of COX-2, pS2,iNOS and TGFalpha were determined by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot. RESULTS: The damage to the surface of gastric epithelium with focal areas of deep haemorrhagic necrosis was induced by repeated WRS. The adaptative cytoprotection against stress was developed with activation of cell proliferation in the neck regions of gastric glands. The ulcer index (UI) in groups II, III and IV was markedly reduced as compared with group I (I: 47.23+/-1.20; IV: 10.39+/-1.18,P<0.01). GMBF significantly decreased after first exposure to WRS with an adaptive increasement of GMBF in experimental groups after repetitive challenges with WRS. After the 4th WRS, the value of GMBF almost restored to normal level (I: 321.87+/-8.85; IV: 455.95+/-11.81, P<0.01). First WRS significantly decreased the expression of pS2 and significantly increased the expressions of COX-2, iNOS and TGFalpha. After repeated WRS, pS2 and TGFalpha expressions gradually increased (pS2: I: 0.37+/ 0.02; IV: 0.77+/-0.01; TGFalpha: I: 0.86+/-0.01; IV: 0.93+/-0.03, P<0.05) with a decrease in the expressions of COX-2 and iNOS (COX-2: I: 0.45+/-0.02; IV: 0.22+/ 0.01; iNOS: I: 0.93+/-0.01; IV: 0.56+/-0.01, P<0.01). Expressions of pS2, COX-2, iNOS and TGFalpha showed regular changes with a good relationship among them. CONCLUSION: Gastric adaptation to WRS injury involves enhanced cell proliferation, increased expression of pS2 and TGFalpha, and reduced expression of COX-2 and iNOS. These changes play an important role in adaptation of gastric mucosa after repeated WRS. PMID- 15526385 TI - Will it make a difference if I show up and share? A citizens' perspective on improving public involvement processes for health system decision-making. AB - INTRODUCTION: Health policy decision-makers are grappling with increasingly complex and ethically controversial decisions at a time when citizens are demanding more involvement in these decision processes. OBJECTIVES: To assess and revise a set of guiding principles for the design of public involvement processes generated from a synthesis of public participation design and evaluation frameworks that can be used to inform the design and evaluation of future public participation processes in the health sector. METHODS: Six focus groups held in five Canadian provinces comprising citizens with considerable experience of public participation processes. RESULTS AND DISCUSSION: Our findings suggest that citizen participants are highly critical of, and discerning about,their public participation experiences. Yet, they are optimistic and determined to contribute in meaningful ways to future public policy processes. They are clear about where improvements are needed and give top priority to what information is shared, and how, among participants and decision-makers. The views of experienced citizens mapped well onto most of the prior principles of public involvement with a few modifications. First, participants gave greater emphasis to the content and balance of information for the purposes of building trust and credibility between citizens and decision-makers. Second, participants viewed themselves, as well as decision-makers, as sources of information to be shared through the consultation process. Finally, participants stressed the importance of getting the information and communication principles right over addressing all other principles. PMID- 15526387 TI - Heirs of liquid treasures. PMID- 15526388 TI - 15th International AIDS Conference. Access for all. The politics of leadership. PMID- 15526389 TI - 15th International AIDS Conference. New drug development. Medications in the pipeline. PMID- 15526390 TI - 15th International AIDS Conference. News on treatment and social issues. Monotherapy, clean needles, and more. PMID- 15526391 TI - 15th International AIDS Conference. News from the Centers for Disease Control. Update on the U.S. PMID- 15526392 TI - In the culture of now. The arrival of the OraQuick rapid HIV test. PMID- 15526393 TI - HIV and aging. PMID- 15526394 TI - Antiretroviral rounds. MSRA: not just for inpatients anymore. PMID- 15526395 TI - Updated guidelines for the use of rifamycins in HIV/TB-coinfected patients. PMID- 15526396 TI - Pharmacological and surgical treatment of obesity. PMID- 15526397 TI - Economic incentives for preventive care. PMID- 15526398 TI - Training to hospital staff to respond to a mass casualty incident. PMID- 15526399 TI - Defending submission-year analyses of new drug approvals. AB - In response to the critique of Mary Olsen, Daniel Carpenter, on behalf of his co authors, addresses the issue of analysis based on the year a new drug is submitted for Food and Drug Administration (FDA) approval, not the year it is approved. Both substantive knowledge of the FDA drug review process and sound social science theory favor submission-year averaging. The history and bureaucratic mechanics of the Center for Drug Evaluation and Review (CDER) conform to the author's assumption. The statistical theory of optimal experimentation also points to the beginning of review as a locus for effects upon decisions. PMID- 15526400 TI - Allergy and eczema: changing patterns, mechanisms and treatment. January 23-24, 2004. Milan, Italy. PMID- 15526401 TI - Selected papers from the Twenty-Fourth Annual Conference on Peritoneal Dialysis. February 9-11, 2004. San Antonio, Texas, USA. PMID- 15526402 TI - Alzheimer's disease, the neuroimmune axis, and viral infection. PMID- 15526403 TI - The occurrence of intercellular bridges during oogenesis in the mouse. PMID- 15526404 TI - [Transfusional therapy of hemodialyzed patient with anti-Cartwright antibodies: case report and review of the literature]. AB - BACKGROUND: Red cell Cartwright antigen (Yta) is very common in the general population therefore patients without red cell Cartwright antigen and with anti Yta alloantibodies due to previous exposure to the antigen are rare. Report about clinical significance of Yta red cell alloantibodies in hemodialysis (HD) patients are scarce. CASE REPORT: We report a cirrhotic uremic patient with anti Yta antibodies who received Yta positive red cells. No adverse reactions nor hemolysis were detected. CONCLUSIONS: We concluded that dialysis patients with anti-Yta antibodies could be safely transfused with blood from Cartwright positive donors. PMID- 15526405 TI - Molecular techniques and the true content of reality. PMID- 15526406 TI - Spotlight on ghrelin in gastroenterology. PMID- 15526407 TI - Retraction. Assessment of the hypothalamic-pituitary-adrenal axis over a 24-hour diurnal period and in response to neuroendocrine challenges in women with and without childhood sexual abuse and posttraumatic stress disorder. PMID- 15526408 TI - [AIDS, a danger without precedent]. PMID- 15526409 TI - Dietary sesame oils inhibits iron-induced oxidative stress in rats [corrected]. AB - The high stability of sesame oil against oxidative deterioration is attributed to lignans in its non-glycerol fraction. The present study evaluates the effects of feeding sesame lignans (sesamin and sesamolin) on Fe2+-induced oxidative stress in rats. Three groups, each of sixteen male weanling WNIN rats, were fed diets containing 200 g casein/kg and l00 g oil/kg (group 1, groundnut oil; group 2, sesame oil; group 3,sesame oil + sesamin (0.4 g/kg). After 45 d of feeding, eight rats from each group were injected with saline (9 g Na Cl/l, controls) intraperitoneally while the remaining eight rats were injected with 30 mg Fe2+/kg body weight as ferrous sulfate in normal saline. The animals were killed after 90 min to evaluate hepatic function and antioxidant status. Compared with those fed groundnut oil (group 1), sesame oil-fed rats(groups 2 and 3) had lower levels of hepatic thiobarbituric acid-reactive substances, serum glutamate:oxaloacetate transaminase activities and serum glutamate pyruvate transaminase activities, indicating protection against Fe-induced oxidative stress. Despite similar tocopherol levels in the three diets, hepatic a-tocopherol levels were higher in rats fed the sesame-oil diets (groups 2 and 3) compared with controls (group 1).However, activities of hepatic antioxidant enzymes (superoxide dismutase and glutathione peroxidase) were significantly (P< 0-05) increased only in rats fed higher levels of lignans (group 3). These observations suggest that sesame lignans may have sparing effects on tocopherols. The increased bioavailability of tocopherols in the presence of dietary lignans might be due to the regeneration of oxidized tocopherols. The synergistic effects of lignans with tocols has nutritional and therapeutic implications. PMID- 15526410 TI - The cause is worse: remeeting Jocasta. AB - This is paper will help bring Jocasta, a figure relatively neglected by psychoanalytic theory, whose lens upon her son has left her to the side, into greater focus. It is not that Jocasta is completely ignored but rather that, when studied, she appears as the dangerous, castrating, forbidden woman to be placated or avoided at all cost. So, although she stands at center stage as the fulcrum of Oedipus's destruction, it is never as a mother longing for her son. The author contends that Jocasta is the personification of an ongoing developmental need on the part of all mothers to separate from their children coupled with a universal longing for reunion. As with Oedipus, Jocasta--a character in a play--is an example of the perverse outcome of forbidden gratifications; but also, as with Oedipus, she is the figurative presentation of normal variations on a theme. PMID- 15526411 TI - On: dream actors in the theatre of memory: their role in the psychoanalytic process. PMID- 15526412 TI - In honor of Vice Admiral James A. Zimble. PMID- 15526413 TI - [Body morphology, coronary risk factor]. PMID- 15526414 TI - [The future of asthmatics]. PMID- 15526415 TI - [Controlling antibiotic resistance]. PMID- 15526416 TI - [5th Nationwide Diabetology Symposium. Diabetes and Gastroenterology. Hradec Kralove, 6-7 July 2003]. PMID- 15526417 TI - Autochronometric abnormalities in patients with cerebral pathology in various locations. AB - Dysfunction of the limbic structures of the temporal lobe of the brain (predominantly the hippocampus), chiasma, hypothalamus, and epiphysis is accompanied by impairment of internal estimation of time periods. In patients with lesions of the temporal lobes and base of the skull, changes in measures of autochronometry were uniform, were the most marked, and were not directly associated with locomotor parameters or with changes in the general functional state of the CNS. In patients with spinal cord or vertebral lesions, including tumors, and in those with extensive lesions of the frontoparietal areas of the neocortex, there were no profound defects in endogenous time estimation, regardless of impairments of the functional state of the synaptic and motor system. These data identify the cerebral oscillatory formations--the hippocampus, the hypothalamic area, and the epiphysis--as structures directly involved in the physiological mechanisms controlling autochronometry. PMID- 15526418 TI - Development of neural stem/progenitor cells from human brain by transplantation into the brains of adult rats. AB - The aim of the present work was to study human neural stem/progenitor cells (SPC) cultured in vitro and their potential to survive, migrate, and differentiate after transplantation into adult rat brain. SPC were extracted from the brains of nine-week human embryos and were cultured in selective medium for three weeks. Transplantation was with suspensions of cells or whole neurospheres; these were studied four weeks after transplantation into the hippocampus, striatum, and lateral ventricles of adult rats. Analysis of transplanted cells was based on various histological and immunohistological staining methods: bisbenzimide, bromodeoxyuridine, and antibodies to human nuclei, vimentin, beta-tubulin, neurofilaments, and glial fibrillar acidic protein, which allowed us to make independent assessments of their state and differentiation. Transplanted SPC from human brains survived well for one month in all areas of adult rat brain without immunosuppression. Cells from suspension transplants migrated intensely and differentiated into neurons and gliocytes. At the same time, transplants of whole neurospheres showed limited or no migration because of the development of a glial barrier. PMID- 15526419 TI - Structure of the sensorimotor area of the cerebral cortex in the offspring of alcoholized rats. AB - Light and electron microscopy and morphometry were used to study the characteristics of the cytoarchitectonics of the sensorimotor cortex and the structures of neurons and their dendrites in 21- and 30-day-old baby rats born to chronically alcoholized females and males. Three categories of morphological changes were identified: signs of delayed maturation of neurons and dendrites, destructive changes to these structures, and signs of repair processes, with dynamics occurring during postnatal ontogenesis. At age three weeks, apart from delayed maturation of neurons and underdevelopment of the dendritic system, there were also spreading destructive changes. Increasing age was associated with increases in repair processes, though destructive changes to neurons persisted, which is evidence for the delayed action of alcohol intoxication of animals on the structural development of the brain in their offspring. PMID- 15526420 TI - Morphofunctional changes in neurons in the temporal cortex of the brain in relation to spatial memory in bulbectomized mice after treatment with mineral ascorbates. AB - The effects of an antioxidant mixture of mineral ascorbates (MA) on the state of neurons in the temporal area of the cortex and the behavior of mice subjected to bulbectomy (BE) were studied; these mice, as demonstrated previously, are characterized by deficiency of spatial memory and the development of a neurodegenerative process in brain structures showing pathological changes in Alzheimer's disease. One month after BE, there were abnormalities in the cytoarchitectonics of the temporal area of the cortex, with loss of clarity of the boundaries between its layers because of dystrophy of pyramidal neurons and foci of loss of these cells. There were sharp increases in the numbers of neurons showing pyknosis, karyolysis, and vacuolysis on the background of decreases in neuronal density. Three weeks of treatment by addition of MA to the diet prevented the degradation of spatial memory in mice after BE and protected neurons in the temporal area of the cortex from degenerative changes. These results provide evidence for the possibility of prophylaxis of neurodestructive changes of the Alzheimer's type. PMID- 15526421 TI - Changes in the activity of NO synthase, energy metabolism enzymes, and the ultrastructure of cerebral cortical neurons in a model of transient ischemia. AB - Experiments were performed on white male Wistar rats under Nembutal-urethane anesthesia. Transient ischemia (10 min) was produced by clamping of the carotid arteries. Some neurons in the parietal area of the cerebral cortex and endotheliocytes in blood vessels showed increases in NO synthesis and in the activity of energy metabolism enzymes (SDH, LDH). Changes in the ultrastructure of some neurons and synaptic formations provided evidence of a state of functional pressure, while changes in others indicated irreversible changes (disturbances in lipid metabolism and destructive changes in the nucleus and cytoplasm). PMID- 15526422 TI - Structural and quantitative characteristics of the dendrites of neurons in the posterior zone of the amygdaloid body in the rat brain. AB - The aim of the present work was to study the structure of the dendritic tree of the main groups of neurons in the posterior zone of the amygdaloid body and to analyze their quantitative characteristics. Frontal sections of rat brain impregnated with Golgi silver nitrate were used to study the characteristics of neuronal organization. Classification of neurons was based on criteria developed by Leontovich (1978) and Polyakov (1973). The main groups of neurons were long axon, rarely branched cells, long-axon, densely branched subcortical cells, and long-axon, densely branched cortical cells. Quantitative studies of seven measures of the dendrite tree showed that the most informative parameters for differences between the main groups of neurons were the number of branching points and the number of free dendrite tips. The contents of rarely and densely branched neuron systems in the nuclei, paleocortex, and intermediate formations are described. PMID- 15526423 TI - Morphofunctional changes in incubated Mauthner neurons in goldfish treated with peptides from scorpion venom. AB - Electron microscopy with negative contrast showed that direct interaction of one of the peptide fractions of scorpion venom with monomeric chromatographically pure actin led to polymerization of actin, transforming it from the globular form to the fibrillar form. The effects of prolonged orthodromic stimulation on the evoked electrical activity and ultrastructure of Mauthner neurons (MN) were studied in incubated slices of goldfish medulla oblongata in the presence of this actin-polymerizing venom fraction. Peptides in this fraction were found to stabilize the amplitude of the electrical response of MN to exhaustion and to protect the ultrastructure of afferent chemical synapses and the neurons themselves from damage induced by stimulation. Enhancements in morphofunctional resistance were accompanied by stabilization of actin-containing specialized synaptic structures--desmosome-like contacts. The data obtained here provide evidence that peptides of this fraction of scorpion venom have direct actions on the actin component of the MN cytoskeleton and demonstrate potential for its use as a pharmacological tool able to penetrate living cells with value for studying the role of actin in the mechanisms of adaptation and memory. PMID- 15526424 TI - Morphofunctional characteristics of spinal cord neurons after single integrative motor loads. AB - Motor and intermediate neurons in the fourth lumbar segment of the spinal cord were studied in 38 mongrel male dogs with the aim of detecting adaptive morphological changes in spinal cord elements after exposure to dosed physical loads (running on the treadmill for periods of 7-35 min). These experiments showed that single integrative physical loads facilitate significant increases in the numbers of functionally active cells. Morphological changes to motor neurons were more marked than those of associative nerve cells. PMID- 15526425 TI - Decreases in glucocorticoid sensitivity as a factor of stress-producing changes in the activity of monoamine oxidase, lipid peroxidation, and behavior in rats. AB - Four episodes of immobilization stress induced a decrease in the sensitivity of rats to glucocorticoid hormones, which was accompanied by anxiogenic behavior, increased MAO-B activity, and a parallel increase in lipid peroxidation (LPO) in brain tissues. There was a simultaneous increase in MAO-B activity in the kidneys and accumulation of LPO products in the liver and kidneys. Administration of Kenalog (2 mg/kg), a pharmacological analog of glucocorticoid hormones, prevented the poststress activation of MAO-B and LPO and decreased the extent of anxiogenic behavioral abnormalities in rats. PMID- 15526426 TI - Functional asymmetry of the frontal cortex and lateral hypothalamus of cats during an operant food-related conditioned reflex. AB - The extent of correlation and the latency of evoked potentials to sound, recorded bilaterally in the frontal cortex and lateral hypothalamus of cats, were studied at different stages of the acquisition of an operant food-related reflex and during sudden transfer to 30% food reinforcement. High correlation coefficients between the evoked responses of the cortex and hypothalamus with left-sided dominance were seen in conditions of high levels of food motivation at the beginning of each experiment and in conditions of the high-probability appearance of the acquired conditioned reflex throughout the rest of the experiment. Comparison of the peak latencies of the early positive (P55-80) components of evoked potentials on the right and left sides showed that shorter latent periods were seen in the cortex on the left side at all behavioral stages, while this occurred (on the left side) in the hypothalamus only when the conditioned reflex was unfixed, while after fixation of the reflex and provision of 30% reinforcement, shorter latent periods were seen on the right side. It is concluded that the high level of left-sided correlation of evoked potentials in the hypothalamus was associated with the motivational and motor components of purposive behavior and was not associated with the emotional tension of the animals provoked by the disruption of the food reinforcement stereotype. PMID- 15526427 TI - From physiological theory to psychological facts (celebrating the 100th anniversary of I. P. Pavlov's Madrid speech). PMID- 15526428 TI - Effects of repeated experience of aggression on the aggressive motivation and development of anxiety in male mice. AB - The sensory contact model allowed aggressive behavior to be formed in male mice as a result of repeated experience of victory in daily social confrontations. In individuals of the low aggressivity, high emotionality line CBA/Lac, repeated experience of aggression led to the development of anxiety, assessed in the elevated cross maze test. Males showed increases in aggressive motivation, measured in terms of increases in behavioral reactions to conspecifics in the partition test. It is concluded that repeated experience of aggression provokes the development of anxiety in male mice and that the level of developing anxiety, like its behavioral correlates, depends on the duration of aggressive experience and the mouse line studied. PMID- 15526429 TI - Duration of retention of changes in the electrical characteristics of command neurons during acquisition of a conditioned defensive reflex in snails. PMID- 15526430 TI - Characteristics of the photoconversion of rhodopsin in the early stages of photolysis. AB - Low-temperature spectrophotometry was used to study the primary stages of rhodopsin photolysis. A digitonin extract of rhodopsin was irradiated at -155 degrees C with blue light of wavelength 436 nm. The stage of the bathorhodopsin - > lumirhodopsin conversion was accompanied by the simultaneous formation of several products. Formation of an intermediate product spectrally similar to the known "blue-shifted intermediate" (BSI) was demonstrated. It is suggested that the appearance of more than one intermediate product at each stage of photolysis reflects the existence of several conformational states of the rhodopsin molecule during its photoconversion. PMID- 15526431 TI - The effects of substance P after central administration on the activity of the mesolimbic system of the rat brain as studied by microdialysis. AB - In vivo microdialysis was used to study the effects of substance P on dopamine, dihydroxyphenylacetic acid, and homovanillic acid levels in the nucleus accumbens in rats. Each animal received sequential injections of physiological saline, 0.1 microg of substance P, and 1 microg of substance P into the lateral ventricle over three days. Dialysates showed increases in dopamine levels in response to neuropeptide, by 41% for the 0.1 microg dose and 71% for the 1 microg dose. The dynamics of these changes also depended on the concentration of the agent. Administration of 1 microg of substance P gave a peak dopamine level at 50 min; the neurotransmitter level remained significantly elevated 75 min after dosage with substance P. The dopamine level was increased only at 75 min when the 0.1 microg dose of neuropeptide was used. Changes in metabolite levels were also dose dependent. After the 1 microg dose, the dihydroxyphenylacetic acid level increased by 28%, while the 0.1 microg dose produced no significant change in the level of this metabolite. The homovanillic acid level did not respond to administration of substance P at either dose. These data support the suggestion that the influence of substance P on the internal compensation system is to a significant extent mediated by dopaminergic mechanisms and provides a possible explanation for the effects of the neuropeptide seen in a conditioned place preference reflex. PMID- 15526432 TI - Mechanisms of disorders of the characteristics of fine movements in long-term hypokinesia. AB - The precision and electromyographic characteristics of single-joint voluntary movements of the human foot, of the programmed and tracking types, were studied, along with the characteristics of rhythmic activity of motor units before and during 120-day antiorthostatic hypokinesia. This latter was accompanied by significant decreases in the precision of the control system, evident as decreases in the number of discriminable force gradations, increases in the absolute and differential thresholds for movements of the programmed type, and sharp increases in the variability of integrated EMG traces from the working muscles during tracking movements. The direction and dynamics of changes in the activity of motor units at different stages of antiorthostatic hypokinesia were different: during the first 14-30 days (stage I), there was a sharp increase in the variability of interspike intervals and an increase in the extent of synchronization of motor unit activity; from day 30 onwards (stage II), there was a reproducible decrease in the duration of interspike intervals, along with disappearance of the synchronization of motor units, while the high level of variability of spike activity persisted. The results obtained here suggest that impairments of precision during stages I and II of antiorthostatic hypokinesia are different in nature and are associated with reflex responses to support unloading at stage I and with atrophic processes in muscles in stage II. PMID- 15526434 TI - New "persona" concept helps site designers cater to target user segments' needs. AB - Using the relatively new "persona" design concept, Web strategists create a set of archetypical user characters, each one representing one of their site's primary audiences. Then, as their site is constructed or upgraded, they champion the personas, arguing on their behalf and forcing the design team to take each audience's needs and wants into account. PMID- 15526435 TI - Creative ways to improve navigation. AB - If your site has voluminous content, you need to find ways to organize the material so that users can find what they need. In its newest upgrade, MayoClinic.com has found a number of good ways to combine massive amounts of content with two- or three-click navigation. PMID- 15526433 TI - Effects of deafferentation on the size and myosin phenotype of muscle fibers on stretching of the rat soleus muscle in conditions of gravitational unloading. AB - The aim of the present work was to assess the contributions of the reflex and local components to preventing decreases in the size and changes in the ratio of fibers containing the slow and fast isoforms of myosin heavy chains during chronic stretching of a postural muscle in rats in conditions of gravitational unloading. A unilateral surgical deafferentation method was used. The results demonstrated that deafferentation of the hindlimb had no effect on preventing reductions in muscle fiber size in conditions of chronic muscle stretching in conditions of gravitational unloading. The results obtained from these experiments did not support the hypothesis that the predominant contribution to preventing the development of atrophic changes comes from activation of muscle afferents in chronic stretching of the unloaded muscle. Deafferentation of both suspended animals and those with normal motor activity led to increases in the proportion of soleus muscle fibers containing the slow isoforms of myosin heavy chain. PMID- 15526436 TI - Improving menu categories. AB - No matter how good a site's navigational tools, site visitors will not use them if the menu categories are ambiguous. Users have to know what to expect when they click on a particular menu item. If the categories are not intuitive, users will have to resort to the site's search engine, ignoring the entire structure. The Pennsylvania Medical Society site (http://www.pamedsoc.org) had been plagued with poor menu labels until it took a step back and improved them. PMID- 15526437 TI - Report on attitudes about personal health records. PMID- 15526438 TI - [Quantitative assessment of the effects of hemispheric asymmetry on the distortion of visual perception of the Jastrow version of the Poggendorff figure]. PMID- 15526439 TI - [Changes in the EEG spectrum and subjective characteristics of the general state after stimulation with variable frequency photic stimuli with two types of organization]. PMID- 15526440 TI - [A neuropsychological and neurophysiological study of memory impairments in patients with left-hemispheric stroke during rehabilitation]. PMID- 15526441 TI - [Psychophysiological correlates of writing and reading difficulties in children of elementary school age]. PMID- 15526442 TI - [Influence of short-term relaxation on the organization of the brain electrical activity of young schoolchildren in the state of quiet wakefulness]. PMID- 15526443 TI - [Metaphorical and associative thinking in healthy children and in children with Asperger's syndrome at different ages]. PMID- 15526444 TI - [Age-related changes in attention and impulsivity in young schoolchildren]. PMID- 15526445 TI - [Central hemodynamic parameters and cardiointervalogram features in infants during early postnatal adaptation]. PMID- 15526446 TI - [Application of chaos theory in analyzing the cardiac rhythm in healthy subjects at different sleep stages]. PMID- 15526447 TI - [Electromyographic characteristics of muscle fatigue in parkinsonian patients exposed to general heating]. PMID- 15526448 TI - [Riding therapy as a method of rehabilitation of children with cerebral palsy]. PMID- 15526449 TI - [Estimation of human biological age based on the parameters of heart rhythmic activity]. PMID- 15526450 TI - [Intermittent normobaric hypoxia as a model of incomplete adaptation]. PMID- 15526451 TI - [Some personality characteristics of fire service specialists under conditions of prolonged intense workloads]. PMID- 15526452 TI - [Nitric oxide correlation with malondialdehyde and angiotensin-converting enzyme in healthy subjects and patients with thoracic wounds]. PMID- 15526454 TI - [Blood rheology, sex hormones, and cortisol in athletes]. PMID- 15526453 TI - [Motor and autonomic asymmetries in athletes with different specializations. Communication I]. PMID- 15526455 TI - [Interrelations between the constitution type and features of muscular activity energetics in sprinters and stayers]. PMID- 15526456 TI - [Homocysteinemia and some other metabolic risk factors of cardiovascular pathology in adolescents with arterial hypotension]. PMID- 15526457 TI - [Characteristics of platelet parameters in newborn infants with intrauterine growth retardation]. PMID- 15526458 TI - [Pharmacological correction of human energy expenditures during immediate adaptation to cold and hypoxia]. PMID- 15526459 TI - [Physiological analysis of features of relaxation achieved by different techniques]. PMID- 15526460 TI - [Ultrasound induced the formation of nitric oxide and nitrosonium ions in water and aqueous solutions]. AB - Nitric oxide, nitrosonium ions, nitrites, and nitrates are formed in water saturated with air under the action of ultrasound. Nitrosonium ions react with water and hydrogen peroxide to form nitrites and nitrates in sonicated solution, correspondingly. Nitric oxide is practically completely released from sonicated water into the atmosphere and reacts with air oxygen, forming NOx compounds. The oxidation of nitric oxide in aqueous medium by hydroxyl radicals and dissolved oxygen is a minor route of the formation of nitrites and nitrates in ultrasonic field. PMID- 15526461 TI - [Distinctive features of the self-association of deoxyhexanucleotide 5' D(GpCpApTpGpC) in aqueous solution: 1H NMR analysis]. AB - The self-association of self-complementary deoxyhexanucleotide d(GCATGC) was investigated in aqueous salt solution. Homonuclear 1H NMR correlation spectroscopy (2D-TOCSY and 2D-NOESY) was used for complete assignments of nonexchangeable protons of the hexamer. The equilibrium reaction constants and thermodynamical parameters of duplex d(GCATGC)2 formation were determined from experimental concentration and temperature dependences of proton chemical shifts of the deoxyhexanucleotide. Distinctive features of the concentration dependences in the range of small concentrations at relatively low temperatures of solution enable one to assume that one single-stranded hexamer sequence forms a compact structure (similar to a hairpin) in aqueous solution. A possible spatial hairpin structure of the hexamer was proposed. Comparative analysis of the experimental and theoretical (using the "nearest neighbor" model) thermodynamical parameters of duplex formation was made. PMID- 15526462 TI - [The formation of liquid-crystalline dispersions of DNA-chitosan complexes under the conditions of molecular crowding]. AB - The formation of liquid-crystalline dispersions from DNA-chitosan complexes in polyethyleneglycol-containing solutions was studied. It was shown that the molecular crowding affects neither the efficiency of binding of chitosan molecules to DNA nor the mode of spatial packing of DNA-chitosan complexes in particles of liquid-crystalline dispersions. PMID- 15526463 TI - [The molecular mobility of water in natural polymers: silk Bombyx mori with a low water content as studied by 1H DQF NMR]. AB - The molecular mobility of water in fibres of natural silk (Bombyx mori) was studied by the double-quantum-filtered (DQF) and single-pulse 1H NMR techniques. The results obtained showed a slow motion of water molecules and their strong interaction with silk macromolecules. At different model functions for resonance lineshape in 1H NMR spectra, the influence of signal linewidth on the estimation of relaxation times and cross-relaxation parameters was considered. The observed 1H DQF NMR signal in B. mori silk fibres (BC = 0.065) indicated a local order and anisotropic motion of water molecules, which leads to 1H-1H dipolar interactions in natural silk fibers due to the creation of the second-rank tensors (T(2,+1), T(2,-1)). DQF spectra were the difference of two Lorentzians with different linewidths and were analyzed using the theory of 1H DQF NMR and the data on residual dipolar interactions in systems with the anisotropic mobility of water molecules. The residual dipolar interactions was insignificant and, as the humidity increased (0.18), no DQF-signals and residual dipolar interactions were observed. PMID- 15526465 TI - [Rotenone-insensitive NADH oxydation in mitochondrial suspension occurs by NADH dehydrogenase of respiratory chain fragments]. AB - Two types of NADH oxidation, rotenone-sensitive and rotenone-insensitive, in suspension of beef heart mitochondria were investigated by the spectrophotometric method. The oxidation of the added NADH by mitochondria in hypotonic media occurs only through the NADH dehydrogenase of the respiratory chain, since it was totally blocked by rotenone or amytal (and also by antimycin A or azide), but the ferricyanide-activated NADH oxidation was insensitive to these inhibitors. The insensitivity of the NADH dehydrogenase to rotenone appears to be due to a shunt of the electron transfer to ferricyanide without involving of ubiquinone. Both types of the oxydation occur through one and the same enzyme, which exists in two states. The evidence in favour of this is that NAD+ and DTT slightly influence the first type of oxidation but strongly inhibit the second one. The ferricyanide activated NADH oxidation takes place in NADH dehydrogenase fragments released from mitochondria. Low Ds-Na concentrations block the respiratory chain NADH oxidation but increase the velocity of the ferricyanide-dependent oxidation. Probably, the increase is the result of the detergent-induced additional releasing of the fragments. The express-method for the preparation of the initially purified fraction with a high yield of detergent-containing fragments of the active enzyme is described. PMID- 15526464 TI - [A study of the effect of irradiation hardness on the properties of the fluoropolymeric matrix modified by polyanionic biologically active substances]. AB - The properties of a synthetic substrate responsible for the behavior of substrate dependent cells in the culture were studied. The effect of the composition of a system water-soluble biopolymer (sodium alginate or methyl cellulose)-synthetic latex SKF-26 and the effect of various types of radiations on its biophysical properties were studied. The results obtained indicate that the addition of water soluble biopolymeric additives to synthetic polymeric films improves the adhesion of cells to the substrate, the adhesion being closely related to the concentration of additives. It was found that the modification methods that determine changes in the charge of the substrate affect the capacity of different cell types for adhesion and proliferation. It was also found that the hardness of irradiation does not affect the vapor permeability and the extent of film swelling. PMID- 15526466 TI - [A network of hydrogen bonds in the reaction centers of Rhodobacter sphaeroides serves as a regulatory factor of the temperature dependence of the recombination rate constant of photooxidized bacteriochlorophyll and primary quinone acceptors]. AB - The dark recombination rate constant for the photooxidized bacteriochlorophyll (P) and reduced primary quinone acceptor (QA) in the photosynthetic reaction centers (RC) from purple bacterium Rhodobacter sphaeroides depends nonmonotonically on temperature. The time of this reaction is approximately 100 ms at 270-300 K and decreases as the temperature both increases and decreases beyond this temperature range. It is known that the dome-shaped dependence of the thermodynamic stability on temperature is an intrinsic feature of many proteins in solution. The experimental results on the nonmonotonous temperature dependence of P+ and QA- recombination rate constant are discussed in terms of general thermodynamic approaches. The dynamic properties of the network of hydrogen bonds that are involved in the relaxation processes accompanying the electron transport are considered as a regulatory factor of the efficiency of electron transfer. PMID- 15526467 TI - [The temperature dependence of high-resolution 1H NMR spectra of Rhodobacter sphaeroides photosynthetic reaction centres in a temperature range of 25-40 degrees C]. AB - High-resolution 1H-NMR spectra registered within a temperature range of 25-40 degrees C revealed a nonmonotonous dome-shaped temperature dependence of the ratio between integral NMR signal intensities determined at ppm intervals 2.5-4.5 and 0.2-2.5 with a maximum at 30 degrees C. This may be due to RC structural changes accompanying the temperature rise and accelerating the recombination reaction between oxidized bacteriochlorophyll and reduced primary quinone at temperatures above 30 degrees C. PMID- 15526468 TI - [Effect of methylmercury chloride on the primary photosynthetic activity of higher plants]. AB - The effect of methylmercury chloride (MeHg) on the fluorescence characteristics of pea seedling leaves and thylakoids isolated from these leaves was studied by the pulse-amplitude-modulation (PAM) fluorometric method. In 3-4 days after the addition of MeHg (20 microM) to the nutritious solution, the maximal (Fv/Fm) and real (under steady state actinic light illumination) (deltaF/F'm) quantum photochemical yield of PS II decreased. The nonphotochemical fluorescence quenching coefficient in control (qN) decreased after its maximum value has been reached. In MeHg-treated samples, this decrease was not observed, possibly due to the disturbance of delta pH energy transducing processes in ATP synthase. This was confirmed by the results of experiments on isolated thylakoids. After MeHg (5 microM) treatment of thylakoids, the photophosphorylation rate and light triggered Mg2+-dependent H+-ATPase activity were suppressed by 20-40%, depending on the duration of MeHg exposure. However, in experiments with isolated thylakoids, no decrease either in the electron transport rate or in the Fv/Fm ratio was observed. In total, the results obtained allow one to assume that MeHg at concentrations and time duration used directly damages the coupling complex. The PS II inactivation in leaves and algae cells may be a result of the oxidative stress processes. PMID- 15526469 TI - [Low radiance in the region of wavelengths 620-660 nm reduces the UV-B-induced damage to photosystem II in spinach leaves]. AB - The effect of monochromatic preirradiation in the region of wavelengths 550-730 nm on the parameters of variable and delayed fluorescence in primary spinach leaves irradiated with UV-B was investigated. It was shown that irradiation of leaves with UV-B at lambda(M) = 300 nm reduced photoinduced changes in variable and delayed fluorescence as well as increased the half rise-time of maximum fluorescence. Preirradiation in the region of 620-660 nm (I = 2-4 W x m(-2)) reduced partly the damage to photosystem II induced by UV-B, whereas preirradiation at lambda(m) = 550, 600, 700, and 730 nm had no significant influence. Potential receptors of red light that take part in the protection of photosystem II against UV-B are discussed. PMID- 15526470 TI - [Damage to erythrocyte membranes induced by high-intensity ultrasound]. AB - The effect of high-intensity ultrasound (11.2-54.2 W/cm2, frequency 36 kHz) on the structural and functional state of erythrocytes was investigated. It was shown that, at short-term action of the ultrasound (up to 1 min), the dose dependent hemolysis of erythrocytes occurs. It was found that the exposure to ultrasound of high intensity (54.2 W/cm2) leads to the disruption of the structural state of erythrocyte membranes, which manifests itself in a change of microviscosity of the lipid bilayer of membranes and inhibition of the activity of the lipid-dependent membrane-bound enzyme acetylcholinesterase. PMID- 15526471 TI - [Fractal properies of gating in potential-dependent K+-channels in Lymnaea stagnalis neurons]. AB - Sets of the channel open times, [tau(o)], and closed times, [tau(c)], and the full set of the channel open and closed times, [tau(o), tau(c)], in the activity of single voltage-dependent K+-channels in mollusc L. stagnalis neurons were analyzed using the rescaled range analysis (Hurst method), fast Fourier and wavelet transforms. It was found that the Hurst dependence for each time series could be approximated by a polygonal line with at least two slopes: H1 and H2 (Hurst exponents). The averaged values of H1 and H2 for the sets [tau(o), tau(c)] were equal to 0.61 +/- 0.03 and 0.83 +/- 0.11, respectively; for the [tau(o)] sets H1 = 0.66 +/- 0.03 and H2 = 0.95 +/- 0.10; for the [tau(c)] sets, H1 = 0.62 +/- 0.05 and H2 = 0.85 +/- 0.10. In some cases, a third slope appeared on the Hurst dependences. It was very variable and ranged between 0.5 and 1. The Hurst exponents H1, H2, and H3 characterized short, intermediate, and long time ranges, respectively. The ranges greatly varied from experiment to experiment. The data obtained show that the channel openings and closings (gating process) represent a persistent process correlated in time. The randomization of the time sets resulted in a single slope, H, of 0.52 +/- 0.02 characteristic of random processes. The results were confirmed by the fast Fourier and wavelet transforms. In addition, possible voltage dependences of Hurst exponents and their correlation with tau(o) and tau(c) were investigated. As a whole, single channel activity may be characterized as a multifractal process with a slight voltage dependence of the Hurst exponents. PMID- 15526472 TI - [The causes of the biological action of electrochemically activated solutions by changes in the growth of Escherichia coli cells]. AB - To study the causes of the biological effect of electrochemically activated solutions, nutrient growth media M 9 were prepared using catholyte and anolyte solutions containing separate components of the nutrient medium, such as distilled water, phosphate buffer, phosphate buffer with chlorides (NaCl, NH4Cl), and chlorides. The biological activity of different nutrient media was assessed by a comparison with the stimulation or inhibition of the growth of Escherichia coli cells in the catholyte and anolyte of the complete nutrient medium M 9. It was shown that medium M 9 prepared on the catholytes of different initial solutions acquired the stimulating properties only if the initial solution contained salts containing chlorine. The stimulating effect of the initial solution was 18-24%. Electrochemical treatment of solutions containing no chlorides (distilled water, phosphate buffer) and subsequent addition of the components of nutrient medium to exposed solutions had neither a stimulating nor the inhibiting effect on cell growth. The cultivation of cells in a nutrient medium based on the catholyte of preliminarily treated hydrochloric acid showed that it is the presence of chlorine ions in solution during electrolysis that causes the stimulating effect of the nutrient medium based on the catholyte. The formation of oxidizers and the inhibitory effect of the anolyte described previously was also observed if the solution contained chlorine ions during electrolysis. Possible mechanisms of the biological effect of catholytes containing chlorides during electrolysis were discussed. PMID- 15526473 TI - [The reconstruction of the dynamics of acetylcholine concentration in synaptic cleft in response to a monoquantum signal]. AB - By using the experimental data and the model of cholinereceptor activation kinetics in frog nervous-muscular junction, the function of acetylcholine release from the nervous terminal and the function of acetylcholinesterase activity in synaptic cleft were reproduced and approximated. These functions can be used in modeling the influence of blockers and other biogenic postsynaptic modulators on synaptic transfer. PMID- 15526474 TI - [A two-compartment model for the dependence of a postsynaptic potential on a postsynaptic current, measured by the patch-clamp method]. AB - As known, the dependence of a postsynaptic potential (PSP)1 on a postsynaptic current (PSC) is not satisfactorily approximated by simple Ohm's law due to a significant role of electrotonic propagation of currents along dendrites. The present work shows that a two-compartment model of a neuron, conjointly solving the two problems of voltage and current clamping, gives quite precisely the PSP on-PSC dependence, in spite of inaccurate reconstruction of currents on dendritic terminals. The two-compartment model is compared with the neuron model consisting of a distributed cylindrical dendrite and a concentrated soma. PMID- 15526475 TI - [Human soleus fibers contractile characteristics and sarcomeric cytoskeletal proteins after gravitational unloading. Contribution of support stimulus]. AB - The effects of support withdrawal and support stimulation on the contractile characteristics of human soleus fibers and cellular factors which influence them were studied. The experimental model of the "dry" head-out water immersion was used in the study. In this model, the hydrostatic pressure on different sites of the body surface are equal so that the experimental conditions are close to the complete supportlessness. A 7-day exposure to dry immersion resulted in a decrease in the maximal isometric tension of the skinned fibers, a decline in the myofibrillar Ca2+-sensitivity, and the relative loss of the titin and nebulin content. A significant decrease in the percentage of fibers containing slow myosin heavy chains was also observed after dry immersion. The application of the mechanical stimulator influencing the plantar support zones with a pressure of 0.2 +/- 0.15 kg/cm2 6 times a day for 20 minutes of each hour brought about a complete prevention of the above listed effects of dry immersion. The data obtained allow one to conclude that the decline in maximal tension and Ca2+ sensitivity as well as myosin shift and loss of sarcomeric cytoskeletal proteins are associated with the support withdrawal during the exposure to dry immersion. PMID- 15526476 TI - [The NMR spin-echo method is used for measurements of the translational water diffusion selectively along the apoplast and the vacuolar and cytoplasmic symplasts of plant tissue]. AB - Simple methods for the registration of translational diffusion of water in apoplast and vacuolar and cytoplasmic symplasts were developed. The methods are based on spin-echo NMR with a pulsed magnetic field gradient and are realized by preliminary inversion of magnetization and the use of paramagnetic doping. It was shown that the diffusion of a part of root water in segments of mais roots is more enhanced than that for the bulk water. The results are explained by the appearance of the rotational movement of the protoplasm and in terms of the hypothesis of water transfer along the apoplast, which balances the cytoplasmic symplast. PMID- 15526477 TI - [A model of resource reallocation during physiological adaptation in Mediterranean fruit fly females]. AB - Analysis of experimental data on longevity and fertility of females of Mediterranean fruit fly Ceratitis capitata demonstrates that flies lay fewer eggs on the average as the average life-span increases. At the same time, the values of individual life-span and the number of eggs laid are positively correlated. To explain these results, a resource model describing the allocation and reallocation of resources between two basic functions, reproduction and maintenance, was proposed. The reallocation of resources, is triggered by changes in environmental conditions such as diet changes, and the parameters of reallocation depend on the current environmental conditions. Modeling demonstrated that the results of experiments are greatly influenced by population selection and heterogeneity. PMID- 15526478 TI - [Spontaneous sector formation in soil streptomycetes colonies]. AB - A model of the formation of sectors during the growth and zone formation of a colony of the soil radiant fungus streptomycetes was developed. It was shown that the basic parameters determining the shape of the sector border are the correlations between the growth rates of the sector of mycelium hyphae and the remainder of the colony and between the frequency and the numbers of branching order of these hyphae. Based on the long-standing observations, a polyseasonal dependence of spontaneous sector formation in streptomycetes zone-forming colonies was demonstrated. PMID- 15526479 TI - [On the regulation of the ecological community structure by a variation of the concentration ratios of resources in environment]. AB - The possibility to regulate the structure of a community using a variational model of the ecological community was studied. The dependences of relative numbers of microorganisms on the initial ratios of resources of nutrients in environment were obtained. The potential possibility of controlling the community structure was shown. Model calculations and experimental data obtained on a community of Chlorococcales indicated that, as the ratios of resources vary, a change of the dominating species takes place. PMID- 15526480 TI - [Visual associative memory and the orientation-contingent color after-effect]. AB - The traditional explanation of the McCollough effect (ME) by selective adaptation of single detectors selective to color and orientation suffers from a number of inconsistencies: 1) the ME lasts much longer (from several days up to 3 months) than the ordinary adaptation, the decay of the effect being completely arrested by night sleep or occluding the eye for a long time; 2) the strength of the ME practically does not depend on the intensity of adapting light; and 3) a set of related pattern-contingent after-effects discovered later required for such an explanation new detectors, specific for other patterns. These properties can be explained, however, in the framework of associative memory and novelty filters. A computational model has been developed, which consists of 1) an input layer of two (left and right eyes) square matrices with two analog receptors (red and green) in each pixel, 2) an isomorphic associative neural layer, each analog neuron being synaptically connected with all receptors of both eyes, and 3) an output layer (novelty filter). The modification of synaptic efficacies conforms to the Hebb learning rule. The function of the model was examined by simulation. After a few presentations of colored gratings, the model displays the ME that is slowly destroyed by subsequent presentations of random pictures. With a sufficiently large receptor matrix, the effect lasts a thousand times longer than the period of adaptation. Continuous darkness does not change the strength of the effect. Like in real ME, the model does not display interocular transfer. The model can account for different pattern-contingent color after-effects without assuming any predetermined specific detectors. Such detectors are constructed in the course of adaptation to specific stimuli (gratings). PMID- 15526481 TI - [On the reactions of the human nervous system to complex-frequency optical stimulation]. AB - A computerized system for precise stimulation and analysis of electroencephalographic (EEG) reactions to two simultaneously presented frequencies of sine-wave light (one constant, 13 Hz, and the other varying from 1 to 6 Hz and vice versa) was used to study the mechanisms of human brain reactivity to complex rhythmical stimulation. The frequencies were generated by computer and presented to the subjects by three different ways: as a result of their simple summation (additively), as a product of their multiplication (multiplicatively, amplitude modulation of constant frequency by the varying frequency), or by separate presentation to different eyes. The dynamics of electroencephalograms for different types of stimulation were compared. Under all three experimental conditions, the dynamics of EEG spectra has demonstrated the same general pattern of resonance activation, which was similar to that observed for the presented signals in the case of their amplitude modulation. Significant positive shifts in the functional state of subjects were observed as a result of stimulation. The results obtained show the leading role of the processes of amplitude modulation in the interaction of integrative, adaptive, and trace mechanisms of the brain functioning during human perception of complex rhythmical stimuli. PMID- 15526482 TI - [Adaptation of the functional system of the living organism to nonregular conjugated actions]. AB - A semiphenomenological approach to the study of the adaptation of a particular functional system of the living organism to nonregular changes in the environmental parameter conjugated with this system was suggested. It was shown that the behaviour of the functional system under these conditions is similar to that of a macroparticle in the rotational Brownian motion. In the adaptation process, a phase of superadaptation at the smallest nonregular actions was revealed. Some examples supporting the occurrence of the superadaptation phase are presented. PMID- 15526483 TI - [Registration of surface fluorescence of endogenous porphyrins during low-level laser therapy of gastric and duodenal ulcers]. AB - With the use of the developed opto-fiber spectroscope and gastroscope, the intensity of fluorescence from the ulcer surface into the gastrointestinal tract was measured in the range of 670-690 nm under the He-Ne laser (= 632.8 nm) illumination. Surface fluorescence data were collected with the use of a special diagnostic fiber with a total diameter of about 2 mm, which combines some illuminating and receiving fibers. To do the measurements, the diagnostic fiber was brought into a slight contact with the mucosa surface. The quantitative results were calculated with the use of the special fluorescent contrast coefficient (Kf), which takes into account the backscattered light as well. After the fluorescent diagnostic procedure, the low-level laser therapy was applied to all patients. It was found that: first of all, during the laser medical cure the registered fluorescence for the patients with a good dynamics of treatment had an evident tendency to a decrease, and the coefficient Kf for observed ulcers tended to approach the value obtained from normal mucosa (Kf), which indicates the normalization of the porphyrin content in the tissue. Second, the retrospective analysis of the efficacy of laser therapy versus initial values of Kf showed that, for the patients with the initial values Kf = 0.2, the efficiency of the laser treatment was the highest: the relation between the patients with good treatment results and bad ones was 4:1 (more than 80%). For the initial Kf = 0.2 0.3, this relation was 3:1 but for Kf = 0.4-0.5 it was 2:3. The differences in the efficiency of the low-level laser therapy, except the cases of the cancer in the ulcers, may be due to different content of porphyrin in the tissue: at high concentrations of the photosensitizer in the ulcer, the effect of the overdosing could be realized. In this case, another technology of laser illumination is needed. The results are discussed in terms of the free-radical conception of the stimulating effect of laser radiation. PMID- 15526484 TI - [Evaluation of glucose utilization in patients with insulin-independent diabetes mellitus by using breathing test]. AB - Commonly used clinical and biochemical parameters, such as the content of glucose, insulin, somatotropic hormone, triglycerides, lactate, pyruvate, and free fatty acids (FFA) in blood of practically healthy subjects and in patients with insulin-independent diabetes mellitus (IIDM), were compared with the parameters obtained by mass-spectrometric analysis of 13CO2 in expired air after 13C-glucose loading. It was shown that, as opposed to healthy subjects, the content of blood glucose and free fatty acids in patients with IIDM increased, the level of glucose dropped in progression upon short-term fasting, and the concentration of lactate changed both upon fasting and after the administration of small test doses of glucose. The use of the 13C-glucose breathing test (13C GBT), which presupposes the loading of safe small doses of glucose enriched in 13C-isotope permitted one to reveal a number of novel quantitative diagnostic criteria for the evaluation of glucose metabolism in patients with IIDM: a decrease in the rate of 13C withdrawal as a constituent of expired carbon dioxide after the administration of 13C-glucose; a reduction in the amount of exogenous glucose metabolized to carbon dioxide; and increased oxidation of endogenous substrates participating in carbon dioxide formation. Small glucose loads proposed by the authors in 13C-GBT are safe for patients with diabetes mellitus and have no effect on the level of blood glucose in healthy persons. The parameters determined by noninvasive 13C-GBT are more sensitive for diagnosis than commonly used biochemical characteristics of blood in patients with IIDM. The diagnostic criteria obtained allow the prediction of the maximum prohibited glucose loading for every patient. PMID- 15526485 TI - [Methodological differences of the physical and computational model approaches]. AB - General principles of modeling, the problem of adequency of models, and the similarity and differences between physical and computational models are discussed. PMID- 15526486 TI - Improving the odds for avoiding dementia in advanced age. PMID- 15526487 TI - Spiritual and emotional needs of bariatric patients. PMID- 15526488 TI - Intracranial hypertension: monitoring and nursing assessment. PMID- 15526489 TI - Pass the salt: indications for and implications of using hypertonic saline. AB - Sodium is the most abundant extracellular ion. Historically, therapy with hypertonic saline was widely used for a variety of conditions. Currently, there are 3 primary indications for its use in critical care: hyponatremia, volume resuscitation, and brain injury. SIADH and CSW syndrome may require sodium replacement, but most cases of hyponatremia can be managed without administration of hypertonic saline. Studies of use of hypertonic saline in hypovolemia and brain injury are promising, but additional research is needed to better define optimal dosing regimens and to determine the relative risks associated with hypertonic saline versus conventional treatment for the management of patients with head injuries and for volume resuscitation in shock states. PMID- 15526490 TI - Determining brain death in adults: a guideline for use in critical care. Mercy Medical Center, Springfield, Mass.. PMID- 15526491 TI - Vasospasm after aneurysmal subarachnoid hemorrhage. AB - Despite every effort, 13% of patients with SAH still die or are permanently disabled as a consequence of vasospasm. Optimal outcome after aneurysmal SAH depends on careful assessment and management of patients throughout the course of hospitalization. Critical care nurses play a crucial role in this process. Efforts continue to understand the cascade of events that lead to cerebral vasospasm and to develop more effective treatments. Many therapies, both traditional and new, are being investigated to reduce the incidence of symptomatic vasospasm and improve the lives of patients who experience this devastating condition. PMID- 15526492 TI - What is the current evidence on pain and sedation assessment in nonresponsive patients in the intensive care unit? AB - Assessing pain and sedation in nonresponsive patients is challenging. A major challenge is the confounding effect of sedation on objective indicators of pain. Clinicians might infer that adequate sedation means different patient states: promotion of amnesia, sleep/rest, patient safety, ventilator synchrony, and hemodynamic stability. Hence, an ideal measure that can adequately address the complexity and individualize the nature of the goals of pain and sedation therapy remains elusive. Furthermore, the behavioral responses to pain and anxiety/agitation (eg, restlessness, ventilator dyssynchrony, and movement) have many similarities. Tolerance to mechanical ventilation has been suggested to have validity in both an ICU pain scale and a sedation scale. Additional research is needed to establish the validity, sensitivity, and specificity of these pain indicators in sedated patients. In the meantime, in circumstances where patients are nonresponsive to external stimuli, clinicians should integrate other information such as actual or potential risks of pain (eg, extensiveness of injury, invasive therapies, intubation) and risks of pain-related functional impairment into their pain assessment in nonresponsive, sedated patients. PMID- 15526493 TI - The fifth element of negligence. PMID- 15526494 TI - Solving the problem of juggling preceptors. PMID- 15526495 TI - Hypertension in obese patients: a dysmetabolic hypertension with a possible adipocyte dysfunction mechanism. AB - Large longitudinal studies showed the epidemiological link between obesity and hypertension. During last years, multiple possible mechanisms involved in this association were identified. Adipose tissue has an important role in the genesis of hypertension in obese patients through several pathways: insulin resistance, leptin, renin-angiotensin-aldosteron system and mediators of inflammation (TNF alpha, IL-6). Adipocyte may be the major player in the development of insulin resistance and hypertension, elements of the metabolic syndrome, responsible for the cardiovascular complications. PMID- 15526496 TI - Arterial compliance in hypertension and heart failure. AB - The purpose of this paper is to investigate the relation between the presence and degree of arterial compliance impairment and, respectively, the clinical conditions of systemic hypertension and/or heart failure. We investigated 19 patients with hypertension, without heart failure (group A), 13 patients with hypertension and heart failure class NYHA II (group B), 10 patients with coronary heart disease and heart failure class NYHA II (group C) and 7 control patients without clinically relevant cardio-vascular diseases (group D). Compliance was evaluated with a Complior device, by measuring carotid femoral and carotid-radial pulse-wave velocities (PWV). Carotid-radial PWV did not differ significantly between the various groups (10.8 +/- 2, 10.6 +/- 0.9, 9.5 +/- 1.5 and 9.9 +/- 1.6 m/s, for groups A, B, C and D, respectively). Carotid-femoral PWV, in group A hypertensive p without heart failure, was 13.1 +/- 2.9 m/s, significantly higher as compared to group C p with coronary heart disease and heart failure (10.5 +/- 2.4 m/s, p = 0.02), as well as compared to group D controls (9.8 +/- 2.6, p = 0.02). Group B p, with heart failure associated to hypertension, had a carotid femoral PWV of 13.5 +/- 3.9 m/s, similar to the one found in group A hypertensive p without heart failure. When comparing groups B and C p with heart failure, we noted that the carotid-femoral PWV was significantly (p = 0.04) increased in hypertensive p, as compared to those with coronary heart disease. In conclusion, the results of our study confirm the impairment of arterial compliance, in hypertensive patients. The carotid-femoral, but not the carotid-radial PWV, were useful for identifying impaired compliance in these patients. Heart failure, associated to hypertension, was not accompanied by a supplemental deterioration of arterial compliance. In fact, our study could not confirm the decrease of arterial compliance, in heart failure patients, as compared to controls. This study suggests the importance of hypertension, with its structural arterial wall changes, in the genesis of arterial compliance impairment. PMID- 15526497 TI - Pulse pressure and inflammatory markers in apparently healthy premenopausal women. AB - Several epidemiological studies showed that pulse pressure (PP), as a marker of large arterial stiffness, is an independent predictor of cardiovascular risk. A high pulse pressure may induce lesions of the vessel walls and of the endothelium of the large arteries. Now, inflammation represents one of the most extensively studied pathways involved in the pathogenesis of cardiovascular disease. The aim of the present study was to evaluate if usual markers of systemic inflammation were correlated with pulse pressure. In fifty-four apparently healthy premenopausal women, without any known disease, we performed anthropometrical and blood chemistry measurements, together with the standard measurements of blood pressure. Mean values of fibrinogen are significantly more elevated in PP > 50 mmHg group than in PP < 50 mmHg group (364.79 +/- 71.07 vs. 329.31 +/- 57.81, P < 0.05). Fibrinogen was also significantly correlated with PP (r = 0.27, P < 0.05). However, after controlling for age the correlation between PP and fibrinogen was not significant (r = 0.18, P = NS). ESR was significantly correlated with PP (r = 0.31, P < 0.05) and systolic blood pressure (SBP) (r = 0.32, P < 0.05) but not with diastolic blood pressure (DBP). After controlling for age the correlation between PP and erythrocyte sedimentation rate (ESR) was not significant (r = 0.20, P = NS), but after controlling for body mass index (BMI) this correlation became significant (r = 0.28, P < 0.05). White blood cell (WBC) was not correlated with age, SBP, DBP, PP, age and BMI and the mean values of WBC were increased in the PP > 50 mmHg group, but not significantly (6637.93 +/- 1615.19 vs. 6416.67 +/- 1427.57, P = NS). In our study we found that ERS and fibrinogen values are related to pulse pressure values. Otherwise, blood pressure values, including pulse pressure, were correlated with the age of the subjects and the correlation between pulse pressure and inflammatory markers (ESR and fibrinogen) was not significant after controlling for age. After controlling for BMI, the correlation between ESR and PP remains significant. PMID- 15526498 TI - Ischemic preconditioning by repeated exercise tests involves nitric oxide up regulation. AB - BACKGROUND: Animal studies show that nitric oxide is involved in delayed ischaemic preconditioning. OBJECTIVES: To determine whether plasma nitrates/nitrites (NOx-, as measure of nitric oxide) are modified by two consecutive effort tests and whether these changes translate into clinical improvement. METHODS: There were studied 22 patients with ischemic heart disease, who performed two effort tests at 24-hour interval. Plasma NOx- level was determined and compared before and after both stress tests. Peak effort, double product at peak effort and maximal ST segment depression were considered clinical end-points and were compared between the two tests. RESULTS: Plasma NOx- increased slightly after the first exercise test compared to pre-test value (17.05 +/- 1.6 vs. 15.38 +/- 1.4 micromol/ml). In turn, after the second test a significant rise of NOx- level (23.65 +/- 2.2 vs. 15.10 +/- 1.3 micromol/ml, p < 0.03) was noticed. The pre-test value was practically identical between the two tests. Peak effort and double product at peak effort remained unchanged between the two tests. Although the ischaemic stress was the same, ST depression was significantly lower (p < 0.01) at the second test (0.85 +/- 0.06 vs. 1.73 +/- 0.16 mm). CONCLUSION: Our study shows an increase of plasma NOx- level after the second of two consecutive exercise stress tests at 24 hour interval, along with a decrease of electrocardiographic consequences of approximately the same ischemic stress. These findings are consistent with experimental data in animals, which point to nitric oxide as both trigger and effector of ischaemic preconditioning. PMID- 15526499 TI - Platelet-activating factor acetylhydrolase activity in patients with chronic stable angina (preliminary results). AB - Coronary atherosclerotic disease is related to endothelial inflammation and dysfunction, thrombosis and plaque instability. Different inflammatory markers are studied in stable angina and coronary acute syndromes, in order to stratify better the risk and to prevent the cardiovascular events. Platelet-activating factor (PAF) and platelet activating factor acetylhydrolase (PAF-AH) represent a complex with proinflammatory actions, possibly related to progression of atherosclerotic lesions. In our study, we investigated PAF-AH activity in 30 patients with stable angina, trying to demonstrate a relation of PAF-AH activity with the severity of the coronary disease. PMID- 15526500 TI - Humalog Mix 25 in patients with type 2 diabetes which do not achieve acceptable glycemic control with oral agents: results from a phase III, randomized, parallel study. AB - BACKGROUND: Humalog Mix 25 (Mix 25) is a premixed insulin mixture of 25% lispro and 75% neutral protamine lispro. Insulin lispro is an analog of human insulin. It is created when the amino acids at positions 28 and 29 of the B-chain of insulin are reversed. The natural sequence in human insulin at this position is proline at B28 and lysine at B29. The pharmacokinetic and pharmacodynamic profiles of insulin lispro indicate that it is more rapid acting, and therefore more physiological mealtime insulin than regular human insulin. OBJECTIVE: Primary objective of this study was to compare twice daily treatment with insulin lispro low mixture (Mix 25) to oral treatment with glibenclamide in patients with type 2 diabetes, with respect to the mean 2-hour postprandial blood glucose excursions after breakfast and dinner. SECONDARY OBJECTIVES: to compare the two treatments with regard to the following: hemoglobin A1c, fasting blood glucose, pre-dinner blood glucose, frequency of hypoglycemia, body weight, treatment satisfaction (by questionnaire). METHODS: The study described is a randomized, open-label, parallel group comparison of two treatment regimens in patients with type 2 diabetes. The study included two periods. The lead-in period lasted 10 +/- 7 days, all patients were taking glibenclamide. The treatment period lasted 16 weeks. Patients were randomized to receive either glibenclamide 15 mg daily or switch to Mix 25 before breakfast and dinner. Study design is illustrated in Fig. 1. Glycemic control was assessed by glycosylated hemoglobin (HbA1c) measurements, 4-point self monitoring blood glucose profiles, and patient reported hypoglycemia. One treatment satisfaction questionnaire (Appendix 1) was completed by each participant. RESULTS: 175 patients were included from the two participating countries (Romania--100 patients and Russia--75 patients). 85 were randomized to receive Mix 25 and 90 to glibenclamide arm. 172 patients were included in the efficacy analysis. Baseline patient characteristics did not show any differences between treatment groups for any of the demographic (age, gender, height, body weight, body mass index) or efficacy parameters (HbA1c or self monitored BG values). The mean age was 59.5 +/- 8.2 years, and 35.5% (61/172) were men. The mean body mass index was 27.2 kg/m2. The mean duration of type 2 diabetes was 10.2 +/- 6.6 years, and the mean duration of sulfonylurea treatment was 5.8 +/- 5.9 years. The mean HbA1c and fasting blood glucose levels were 10.07 +/- 1.4% and 11.6 +/- 2.8 mmol/L, respectively, in the glibenclamide group and 9.85 +/- 1.2% and 12.2 +/- 2.9 mmol/L, respectively, in the Mix 25 group. At the end point, all efficacy parameters were better improved in Mix 25 group (HbA1c, fasting blood glucose, 2-hour postprandial blood glucose). Mean HbA1c was significantly lower in the Mix 25 group than in the GB group (Mix 25, 8.5% +/- 1.3%; GB, 9.4 +/- 1.8%; P = 0.001). For all self-monitored blood glucose values (Fig. 2) a larger decrease from baseline was observed in the Mix 25 group: -1.4% versus -0.7% for HbA1c, (P = 0.004); -2.8 mmol/L versus -1.1 mmol/L for fasting blood glucose, (P < 0.01); -5.1 mmol/L versus -1.7 mmol/L for the morning 2-hour postprandial blood glucose, (P < 0.001); -2.2 mmol/L versus -0.8 mmol/L for the evening preprandial blood glucose, (P < 0.05); and 4.4 mmol/L versus -1.5 mmol/L for the evening 2-hour postprandial blood glucose, (P < 0.001). Percentage of patients experiencing at least 1 episode of hypoglycemia was--as predicted- higher in the Mix 25 group (44.7% versus 10.3%; P = 0.01). Patients expressed more satisfaction with Mix 25 than with GB, as measured by the weighted combined score on a treatment satisfaction questionnaire (2.0 +/- 1.3 vs 0.7 +/- 1.3). CONCLUSIONS: When glycemic control can no longer be achieved by oral antidiabetic agents, treatment with insulin should be considered as the next therapeutic option. Mix 25 provided good overall glycemic control, as well as patient treatment satisfaction. PMID- 15526501 TI - Erectile dysfunction in diabetes mellitus. AB - Erectile dysfunction (ED) is 2-3 times more frequent in men with diabetes mellitus than in men without such a history and might be an early marker of endothelial dysfunction. We studied a group of 310 unselected male patients within the Clinical Center of Diabetes and Metabolic Diseases of Dolj County, with ages ranging between 20-78 years (57.43 + 0.835) and a positive history of diabetes mellitus for 1-47 years (10.09 +/- 8.715). Erectile dysfunction, quantified using SHIM (Sexual Health Inventory for Men), was present in 196 patients (63.2%); severe in 52 patients (16.8%), moderate in 42 patients (13.5%) and mild in 102 patients (32.9%). Erectile dysfunction showed a positive correlation with age after 65 years, history of diabetes of more than 10 years, obesity, stroke, arteriopathy, retinopathy, neuropathy and the smoking habit and was not correlated to the type of diabetes mellitus, history of diabetes less than 10 years, diabetes therapy, hypertension, ischemic heart disease, nephropathy, dyslipidemia and alcohol consumption. Our results plead for a holistic approach of the diabetic patient, irrespective of age, in order to detect and to treat all the risk factors, keeping in mind that the appearance of erectile dysfunction might indicate the presence of occult chronic diabetes complications. PMID- 15526502 TI - Longitudinal study of the FEV1, cumulative exposure to dusts and silicosis in gold miners. AB - OBJECTIVES: The aim of this longitudinal study was to assess the rate of decline in FEV1 (deltaFEV1) in gold miners with silicosis and to relate these findings with the radiographic picture, age, smoking habits and cumulative exposure to dusts. METHODS: The study included 73 gold miners with silicosis and 73 miners exposed to dusts but without silicosis. The clinical, radiological and functional assessment was made at least twice in each miner at the beginning and at the end of an adequate interval of the investigation which was extended for a mean period of 38.9 months for the group with silicosis and 33.6 months for the group without silicosis. RESULTS: In the group of miners with silicosis deltaFEV1 was much higher (187 ml/year) as compared with the mean value found in the miners without silicosis (43.2 ml/year). No significant differences were found for deltaFEV1 according to the age criterion (in both groups) and to their smoking habits (in the group with silicosis). In silicosis, a very good correlation exists between the index of cumulative exposure to dusts and the magnitude of the decline rate of FEV1. In the miners, group without silicosis, deltaFEV1 had not any significant correlation with cumulative exposure to dusts. CONCLUSIONS: Our observations revealed that the cumulative exposure to dusts affected the decline rate of FEV1 only to the extent to which it determined silicosis and intervened less in the functional decline in miners without silicosis. Smoking habits increased AFEV1 only in miners without silicosis. PMID- 15526503 TI - Study of anti-cardiolipin and anti-beta2-glycoprotein I antibodies in patients with ischemic stroke. AB - The study of IgG and IgM anti-cardiolipin antibodies (IgG/IgM aCL) is now well accepted and is routinely used in the risk assessment of various conditions associated with thrombosis. The aim of the study was to define whether the investigation of aCL is sufficient by itself to evaluate a risk of ischemic stroke. Frequency of aCL and anti-beta2-glycoprotein I (beta2-GPI) antibodies was prospectively investigated in 96 patients with ischemic stroke and in 119 controls by ELISA. In ischemic stroke patients IgG aCL were found in 36%, the IgM aCL were found in 58%, the IgG-IgM-aCL were found in 43%. The levels of both antibodies were higher in patients with ischemic stroke than in controls (p < 0.01). In controls, IgM-aCL were positive in 2% and IgG-aCL antibodies were negative. IgG-beta-GPI Abs were found in ischemic patients in 19% and IgM-beta2 GPI Abs in 37%. The IgG-IgM-anti-beta2-GPI Abs were found in 24% patients. They were negative in controls. There was a correlation between levels of aCL and anti beta2-GPI Abs for both isotypes (r = 0.728) but not between IgG- and IgM-beta2 GPI Abs. IgG-aCL test was more sensitive for ischemic stroke than the IgG-beta2 GPI Abs test (71.4%, respectively 65.7%) but less specific (66.8%, respectively 88.6%). The sensitivity of anti-beta2-GPI Abs for ischemic stroke was increased when both isotypes were tested. These results showed that aCL and anti-beta2-GPI Abs could be pathogenetically important for ischemic stroke and that anti-beta2 GPI Abs testing might contribute to a better evaluation of ischemic stroke. PMID- 15526504 TI - Chronic hepatitis C virus infection mimicking rheumatoid arthritis. AB - We present the case of a young female patient diagnosed two years ago with rheumatoid arthritis (AR) for which she is taking methotrexate (MTX), who develops cutaneous lesions highly suggestive of porphyria cutanea tarda, diagnosis confirmed by biochemical means. It is noteworthy that she was regularly taking oral contraceptives until the moment of appearance of the skin lesions. The association of those two illnesses, particularly in the case of MTX treatment can raise some problems regarding the potential direct causality relationship. This is why we tried a new diagnostic hypothesis: is chronic hepatitis C virus infection, hypothesis that we verified by means of the presence of anti-VHC and of RNA-VHC. It is well known now the association between chronic viral C infection, rheumatoid syndrome and porphyria cutanea tarda (PCT). The latter are extrahepatic manifestations of that viral infection, thus representing a major indication for antiviral treatment. Our patient received that treatment and she had a very good outcome of the skin lesions. We suggest that the differential diagnosis of any arthritis should always comprise chronic hepatitis C viral infection. PMID- 15526505 TI - Current indications for metformin therapy. AB - Metformin (dimethyl-biguanide) is an oral antidiabetic drug, which decreases hepatic glucose production (gluconeogenesis) and increases peripheral glucose uptake by muscles. Metformin is a first-line drug in the treatment of overweight and obese type 2 diabetic patients, offering a selective pathophysiological approach by its effect on insulin resistance. It has been shown in a number of studies to improve clinical outcomes in type 2 diabetic patients. It has been demonstrated in a number of studies that metformin has multiple biological effects - it has been shown to have platelet antiaggregating effects, to reduce the rate of formation of advanced glycation end products (AGEs) and to decrease the cellular oxidative reactions, thus demonstrating the antioxidant effects of the drug, which may largely explain its vascular protective effect. A number of studies have established the favorable effect of metformin on body weight, insulin resistance, hyperinsulinaemia, lipid parameters (total cholesterol, HDL cholesterol, LDL-cholesterol, triglycerides), arterial hypertension, fibrinolysis, endothelial dysfunction. Thus metformin appears to have a broad set of pharmacological properties, making the drug potentially applicable even in nondiabetic situations such as obesity, extreme insulin resistance with acanthosis nigricans, polycystic ovary syndrome, etc. Metformin has been demonstrated in the Diabetes Prevention Program to be a drug with great potential in preventing the conversion of IGT to type 2 diabetes. Thus, metformin appears to be a drug with multiple therapeutic effects far beyond its effect on lowering blood glucose in diabetes mellitus. PMID- 15526506 TI - Restenosis in the coronary stent is an argument for the surgical myocardial revascularization. AB - BACKGROUND: This study will evaluate the difference between the frequency of restenosis in myocardial revascularization procedures by stents and CABG by coronarographic control after clinical criteria (angina). METHODS AND RESULTS: Out of the total of 6564 coronarographies performed (1999-2002) for diagnosis purposes, 3110 patients (44.8%) underwent myocardial revascularization procedures, PCI or CABG. PCI was performed in 981 patients (31%) and CABG in 1148 patients (37.3%). At the same time, we performed in our units 2067 surgical procedures, out of which 1148 (55%) revascularizations by CABG. The angiographic control for patients with myocardial revascularization by stent (55 patients) or CABG (50 patients) was performed by clinical criteria (angina reappears) on 105 patients. The restenosis we found in 47 patients (74.5%) treated by stent revascularization and in 29 patients from CABG procedures. In our study restenosis rate was 4.8% in patients with PCI and 2.5% in patients with CABG. CONCLUSIONS: Both surgical (CABG) and percutaneous coronary artery revascularization (PCI) have proved to be extremely effective in the treatment of patients with multivessel coronary disease. In our study restenosis rate was smaller in the patients with CABG than in the PCI group, taking into account the fact that we did not use drug eluting stent (DES) on a large scale. Results from the series of randomized trials (SOS, SIRIUS, ERACI, ARTS, BARI, etc.) have shown that the restenosis phenomenon is an apparent advantage in patients with DES. Restenosis prevention is a complex phenomenon (inflammation, procoagulation, cellular migration, etc.) and DES appearance opens a new era in PCI. PMID- 15526507 TI - Rovamycine as add-on treatment in unstable angina and 4 year evolution with major cardiovascular events. AB - BACKGROUND: Major antibiotic trials targeting Chlamydia Pneumoniae or the pathogen burden in acute coronary syndromes reported conflicting data. Only a minor impact of antibiotic treatment on major cardiovascular events (MACE) incidence was demonstrated in some studies. METHODS AND RESULTS: 109 unstable angina patients were randomised in: group C receiving conventional treatment, group R treated with Rovamycine 12 days 4.5 MUI iv /day as add-on therapy, group R1 treated with Rovamycine 12 days 4.5 MUI iv/day followed by 6 MUI/day per os for another 12 days add on treatment. Randomisation into the therapeutical groups was independent of the serological status for Chlamydia pneumoniae. The primary adverse end-points of the study were the incidence of major cardiovascular events at 3 months, 6 months and at 4 years and the 4 years cumulated end-point rate. Secondary adverse end-points were the incidence of recurrent stable angina at 3 and 6 months and the incidence of increased serum levels of C reactive protein and fibrinogen at 3 and 6 months. Statistics used multiple regression analysis and Chi square test. At 6 months the incidence of unstable angina with readmission was significantly lower in groups R and R1 compared to group C (p < 0.001, respective p < 0.0001) and significantly lower in group R1 compared to group R (p < 0.0001). The incidence of nonfatal myocardial infarction at 6 months was significantly lower in groups R and R1 compared to group C (p < 0.0001). The incidence of cardiovascular death was significantly lower in group R1 compared to group C and R (p < 0.001). At 4 years the incidence of unstable angina with readmission and the cumulated end point rate were significantly reduced in groups R and R1 compared to group C. The 3 months incidence of increased serum levels of C reactive protein was significantly decreased in group R1 compared to groups C and R (p<0.001). The 3 months incidence of increased serum levels of fibrinogen was significantly lower in groups R and R1 compared to group C (p<0.002, respectively p<0.001). CONCLUSIONS: In patients with unstable angina Rovamycine as add-on treatment to the conventional treatment lead to a significant decrease of MACE incidence at 6 months and to a significant decrease in the 4 years incidence of unstable angina with readmission. The beneficial effect of Rovamycine was parallel to the decrease in serum inflammations markers concentration. PMID- 15526508 TI - The effects of carvedilol a nonselective beta-blocker on portal hemodynamics in cirrhosis. AB - Portal hypertension is the result of increased hepatic resistance and portal influx. AIM: To assess the effects of Carvedilol, a 3rd-generation nonselective beta blocker with alpha1-adrenergic activity on portal and systemic homodynamic in patients with cirrhosis and portal hypertension. METHODS: Fifty patients with cirrhosis and portal hypertension were divided into two groups and statistically compared as follows: group I - 25 patients received Carvedilol, 12.5 mg/day and group II - 25 patients received placebo for six days. All the patients had hemodynamic, endocrine and renal measurements before and after administration of Carvedilol or placebo. They underwent creatinine clearance, lithium clearance, plasma renin activity, concentration of plasma aldosteron and urinary sodium excretion. Hemodynamic effects were assessed by portal flow volume and velocity, cardiac output and medium blood pressure. RESULTS: Carvedilol significantly increased the portal blood flow and velocity (p<0.05). Carvedilol reduced the medium blood pressure (p<0.001) with statistically insignificant alterations in creatinine clearance and 24-hours urinary sodium excretion. Carvedilol also reduced the concentration of plasma aldosteron (p < 0.002). CONCLUSIONS: Carvedilol can be used as an alternative drug for the prophylactic treatment of portal hypertension with careful monitoring of blood pressure regarding its hypotensive effects. PMID- 15526509 TI - Correlation between the immunophenotypical presentation of dendritic cells and the clinical response to anti-rheumatic treatment in rheumatoid arthritis. AB - BACKGROUND: Dendritic cells (DCs) are potent antigen-presenting cells (APC) that are deeply implicated in the initiation and exacerbation of rheumatoid arthritis (RA). Active RA is associated with an activated DCs population as demonstrated by high expression of adhesion and co-stimulatory molecules. PURPOSE: To compare the expression of adhesion and co-stimulatory molecules on DCs from synovial tissue (ST) in patients (pts) with RA and the clinical status before and after treatment with disease modifying antirheumatic drugs (DMARDs). METHODS: Samples of ST were obtained from RA patients at the time of hip or knee replacement or arthroscopy. Clinical status (assessed by the American College of Rheumatology - ACR - core set and the DAS28) and co-stimulatory and adhesion molecules on DCs were evaluated before and after treatment. Immunophenotype of DCs was analyzed by two color immunofluorescence microscopy. RESULTS: In patients with active RA we found a highly differentiated subpopulation of DCs that expressed an activated phenotype. After treatment, the expression of adhesion and co-stimulatory molecules on ST DCs was correlated with the ACR and DAS28 clinical response. CONCLUSION: Clinical outcome and the immunophenotypical presentation of ST DCs after DMARDs treatment are closely correlated in pts with RA. The co-stimulatory activity in the synovium is important in determining the course of the disease and provide new therapeutic targets for immune intervention. PMID- 15526510 TI - Metformin in the treatment of obesity in subjects with normal glucose tolerance. AB - The aim of the present study was to evaluate the effect of metformin on body weight, body fat mass, waist circumference, lipid parameters, insulin secretion and insulin resistance in obese subjects with normal glucose tolerance. 26 subjects (16 females and 10 males), mean age 37.3+/-9.7 years and mean BMI 32.3+/ 9.1kg/m2 were treated with individual hypocaloric diet and metformin at a mean dose of 2.38+/-0.32g for six months. Insulin secretion was studied during OGTT; insulin resistance was assessed by the HOMA index. We have found a reduction of 3.24% in body weight, which was accompanied by a decrease in body fat mass (by 7.45%, p<0.01) and in waist circumference (p<0.01). Insulin resistance decreased by 14.5% (p=0.02) after metformin. There was a significant decrease in the AUC for the total (by 17.6%, from 6999+/-1453 to 5763+/-1511mU/L.120min, p<0.01) and stimulated insulin secretion (by 17.1%, from 4215+/-1011 to 3495+/ 925mU/L.120min, p=0.01) during OGTT after six-month treatment with metformin. In conclusion, the results from the present study demonstrate that metformin contributes to a reduction in body weight, body fat mass and waist circumference, improves insulin sensitivity and decreases basal, total and stimulated insulin secretion in obese subjects. Thus metformin appears to be an effective and well tolerated drug in the treatment of obesity in subjects with normal glucose tolerance. PMID- 15526511 TI - IgG anti-cardiolipin antibodies-markers of inflammation in diabetic patients with ischemic stroke. AB - Some researchers investigated the role of anticardiolipin antibodies in the pathogenesis of the ischemic stroke. The objective of the study was to evaluate the relationship between the prevalence of the IgG anti-cardiolipin antibodies (IgG aCL) and ischemic stroke in patients with diabetes mellitus. IgG aCL were prospectively investigated in 87 diabetic patients with ischemic stroke (mean age 78.5 years) and in 68 diabetic patients without ischemic stroke (mean age 72.8 years). IgG aCL were determined by ELISA. Twenty-three out of 87 diabetic patients without ischemic stroke had IgG aCL and 25 out of 68 diabetic patients without stroke had IgG aCL. The IgG aCL titer did not differ significantly between patients with ischemic stroke and those without stroke (p>0.05). Our data showed that IgG aCL are not a risk factor for ischemic stroke in diabetic patients. Both patients with diabetes and those with diabetes and ischemic stroke had serum IgG aCL. IgG aCL indicate inflammatory conditions in these pathologies. PMID- 15526512 TI - The effect of fasting on the parameters of the antioxidant defence system in the blood of vegetarian human subjects. AB - OBJECTIVES: Although the evidence is strong that fasting has anti-tumor, anti inflammatory and anti-ageing actions, the mechanisms responsible for these phenomena are still unclear. An ameliorated antioxidative defence with fasting may be the cause of such effects. The objective of the present work was to determine the influence of fasting on antioxidant systems in healthy young vegetarian humans. DESIGN AND METHODS: We measured Trolox Equivalents Antioxidant Capacity (TEAC) of plasma, erythrocytes superoxide dismutase (SOD) activity, blood glutathione peroxidase (GPx) activity, level of total blood non-proteic thiols (TBNT), plasma ceruloplasmin activity, plasma level of NO metabolites (the sum of nitrites and nitrates, NOx), in 18 healthy young humans (age 20-27 years) after 12h (overnight fasting) and 80h of fasting. RESULTS: Trolox Equivalents Antioxidant Capacity of plasma, the level of total blood nonproteic thiols, plasma ceruloplasmin activity and plasma concentration of nitrites and nitrates were significantly increased after 80h of fasting. Superoxide dismutase activity and glutathione peroxidase activity were lower after 80h of fasting. CONCLUSIONS: Our results suggest that fasting induces the "reorganisation" of antioxidative defence lines: fasting increases especially plasma protective systems (total antioxidant capacity of plasma, plasma ceruloplasmin activity) and decreases an erythrocytes antioxidant enyzme (superoxide dismutase) and blood glutathione peroxidase. PMID- 15526513 TI - Effect of kanamycin and gentamicin on frog n. sciatic--m. gastrocnemian preparation. AB - The effect of kanamycin and gentamicin on frog n.sciatic-m.gastrocnemian preparatus was analyzed. Both substances decreased the muscular contraction in both after direct electrical stimulation of the muscle and electrical nerve stimulation, in a dose-dependent manner. Both substances were used at a concentration near the therapeutical concentration in human. The effect of kanamycin was more important than the effect of gentamicin. These data obtained in presented experimental condition are in accord with clinical observation but are not in accord with the possibility that aminoglycoside antibiotics block presynaptic Ca2+ channels. PMID- 15526514 TI - Kikuchi's disease associated with systemic lupus erythematosus and autoimmune like hepatitis. AB - The authors present a case of Kikuchi's disease associated with systemic lupus erythematosus (SLE) and autoimmune-like hepatitis. Kikuchi's disease, or histiocytic necrotizing lymphadenitis is occasionally associated with SLE and mildly elevated aminotransferases. A 17-year old woman presented with fever, arthritis and bilateral cervical adenopathy. Histopathological and immunohistochemical examinations of an excised lymph node showed evidence of Kikuchi disease. An elevation of hepatocytic enzymes (aminotransferases and gamma glutamyl transpeptidase) associated with smooth muscle antibodies in a titer of 1/320 was present. Clinical symptoms and laboratory tests improved after "pulse" corticotherapy. The association described has not been described in the literature, but is considered possible due to the immune pathogenicity of the 3 simultaneous diseases. Kikuchi-Fujimoto's disease, or histiocytic necrotizing lymphadenitis (HNL), is a rare condition first described in 1972 independently by Kikuchi and Fujimoto. HNL is rarely associated with systemic lupus erythematosus (SLE). It is a benign illness characterized by fever and cervical adenopathy and has a self-limiting course. PMID- 15526515 TI - Relapsing polychondritis with splenic abscess. AB - A patient aged 42, diagnosed with polycondritis for approximately 14 years is presented; she has undergone urgent surgery for a splenic abscess in imminent fistulization in the left pleural cavity. Her susceptibility to infections is marked in time by surgical interventions for pultaceous amygdalitis, an abscess of the right submandibular salivary gland, a splenic abscess. To be noted the peculiar connection between the illness and the pregnancy, which differs from the data to be found in reference literature that is the association with a tendency to spontaneous abortion and the sudden installation of an evolutionary acute episode during pregnancy, which was followed by deafness. Based on these facts, immunopathogenic observations on recurrent polycondritis are getting into shape. PMID- 15526516 TI - Multiple myeloma in remission complicated by bone marrow granulomas. AB - BACKGROUND: Granulomas are encountered in 1-2% of biopsies performed in various hematological and non-hematological diseases. Almost 50% of bone marrow granulomas are associated with infections and 25% with hematologic disorders, especially lymphoma and multiple myeloma. Toxoplasmosis is reported to induce granulomas in bone marrow inmunosuppressed patients. On the other side, long-term unexplained remissions after conventional treatment in multiple myeloma were mentioned in up to 10% of cases. CASE REPORT: A 56-years-old female patient was diagnosed with IgG(kappa) multiple myeloma in 1992. After 5 years, being still in complete remission, frequent bone marrow epithelioid non-caseating granulomas were noticed in biopsy, without clinical symptomatology or modifications of routine paraclinical examinations. The history revealed no treatments with antiarrhythmic, antihypertensive, anticonvulsivants or nonsteroid antiinflammatory drugs. The serologic tests for other infections or systemic diseases known to induce granulomas were negative, except those for toxoplasma gondii IgG. The treatment with azithromycine and pyrimethamine induced the disappearance of granulomas, simultaneously with an important decrease of anti toxoplasma IgG antibodies titer. CONCLUSIONS: The bone marrow granulomas provide a valuable histologic clue to opportunistic infections and the bone marrow biopsy is useful for their diagnosis. In the specific case of toxoplasmosis, a recently proposed treatment with azithromycin induced the resolution of the granulomas. Due to the usual lack of specificity of the most bone marrow granulomas, a broad and long-term clinical, histopatological and serological follow-up to establish the etiology should be performed. PMID- 15526517 TI - Trend in randomized controlled trials. AB - Virtually any study may achieve statistical significance if we increase the sample size indefinitely. We cannot discuss about trend without interpreting confidence intervals (CI). The sample size being computed function of the magnitude of the clinical effect is considered worth detecting, when the result is not statistically significant, it is neither clinically significant. Increasing the sample, the "trend" can disappear or even change its direction. PMID- 15526518 TI - Coagulation factor XIII and atherothrombosis. A mini-review. AB - Coagulation factor XIII is a transglutaminase catalysing the crosslinking of fibrin chains as well as the formation of covalent links between several extracellular matrix proteins such as fibronectin, vitronectin and collagen. By mediating the incorporation of alpha2 antiplasmin into the fibrin network, this factor also interferes with fibrinolysis. Increased plasma factor XIII activity was reported by our laboratory 30 years ago in hypertriglyceridemic subjects who also displayed increased activity of serum cholinesterase, a marker of hepatic protein synthesis, and a delayed diluted, blood clot lysis time. Recent data in the literature emphasize a relationship between insulin resistance (metabolic syndrome) and increased plasma levels of factor XIII, confirming our results. It was also reported that a faster activation of this factor related to the Val 34 leu polymorphism provides protective effect against myocardial infarction and stroke, this effect being however negated in patients with insulin resistance and high plasma levels of plasminogen activator inhibitor-1. The pathogenic role of factor XIII in atherothrombosis seems to be bivalent. On the one side, an increased activity would favor the persistence of fibrin depositions and increase plaque burden, while on the other side it would reduce plaque vulnerability and the risk of downstream embolization. PMID- 15526519 TI - Subclinical atherosclerosis, carotid intima-media thickness and the cardiovascular risk. AB - The refinement and the improvement of the cardiovascular risk factors assessment became one of the high priorities in the global approach of cardiovascular diseases. Recent studies showed that increased carotid intima media thickness (IMT) is a relatively simple noninvasive marker of the global atherosclerotic disease, even in the asymptomatic stage. There is a strong link between IMT and both traditional and emerging cardiovascular risk factors, and IMT also predicts the occurrence of major cardiovascular events. Thus, IMT, as a marker of subclinical atherosclerosis, might improve the risk assessment in asymptomatic patients at risk. PMID- 15526520 TI - Pulmonary involvement in diabetes mellitus. AB - Diabetes mellitus involves the lungs in the course of the complex phenomena it generates. Recent research in animal diabetes as well as in human diabetes demonstrated biochemical changes at the pulmonary level such as the suppression of anyline p-hydroxilase, the reduction of the activity of glutathione peroxidase, the development of NO-dependent endothelial dysfunction, microsomal disorders, increased heparan sulphate at the level of the vascular basement membrane, increased levels of advanced glycation end-products and the derangement of bronchial mucus production by amyline. Structural modifications of the lung parenchyma were observed such as the narrowing of the alveolar space, the flattening of the alveolar epithelium and the expansion of the interstitium. Aside from the involvement of the pulmonary vessels there is the involvement of the basement membranes of the alveolar epithelium, the bronchial epithelium and the pulmonary capillaries. The consequences of local oxidative stress, the increased vascular permeability and the modifications in mucus secretion lead to the reduction of pulmonary volumes, pulmonary diffusion capacity, elastic recoil with involvement of restrictive lung disorders, diminished bronchial reactivity and diminished bronchodilatation. Data of pulmonary pathology obtained from patients as well as pulmonary involvement of children born of diabetic mothers are presented succinctly. PMID- 15526521 TI - Therapeutic efficiency of continuous renal replacement therapy--experience of a single Romanian center. AB - We studied 24 patients with acute renal failure (ARF) or acutization of chronic renal failure (CRF) caused by a variety of different disorders. A rapid decline in renal excretory function, fluid, electrolyte, and divalent ion disturbances determined us to initiate continuous renal replacement: continuous venovenous hemofiltration (CVVHF) - 28 procedures in 15 patients and continuous venovenous hemodiafiltration (CVVHDF) - 11 procedures in 11 patients. The mean duration of CVVHF was 21.6+/-6.9 h, with a mean blood flow rate of 116.9+/-16.4 ml/min and an ultrafiltration rate of 6.4+/-4.6 ml/min. The serum creatinine level decreased from 12.6 to 8.3 mg% and the concentration of urea from 237 to 166 mg%. The mean duration of hemodiafiltration was 24+/-8.5 h, with a mean blood flow rate of 134+/-15.2 ml/min, a mean dialysate flow of 35+/-7 ml/min and a mean ultrafiltration rate of 5.6+/-2.1 ml/min. The serum creatinine level decreased from 11.6 to 6.36 mg% and the concentration of urea from 236 to 137 mg%. Survival rate was 79.2% (19/24 patients). The complete recover of renal function was achieved in 5 patients, partial recover in 5 patients and 9 patients were included in chronic dialysis. In 8/24 (30%) patients we encountered complications, such as hypotension in 2 cases, bleeding disorders in 5 cases or diselectrolithemias in 2 cases. As a consequence, continuous renal replacement therapy is efficient, having an acceptable rate of adverse effects in patients with ARF or acutization of CRF. PMID- 15526522 TI - Double bolus of 0.75 MU streptokinase plus enoxaparin versus front-loaded alteplase plus unfractionated heparin in ST-segment elevation myocardial infarction. AB - OBJECTIVE: To compare the efficacy and safety of an accelerated streptokinase regimen (double bolus of 0.75 MU in 10 min) in combination with enoxaparin (SK0.75Enox regimen) with the one of the front loaded alteplase (t-PA 100 mg/90 min) plus heparin (the t-PAHep regimen) in patients (pts.) with ST-segment elevation acute myocardial infarction (STAMI). METHODS: One hundred seventy three pts. (age 18-74) treated within the first 6 hrs. after the onset of STAMI with the above two mentioned thrombolytic regimens were included. 1. The group SK0.75Enox (102 pts.) received an i.v. bolus of 40 mg Enox followed by 0.75 MU SK in 10 min. A second bolus of 0.75 MU SK would be administrated only if no bed side signs of coronary reperfusion (CR) were detected within the next 50 min. After thrombolysis Enox was administered 1 mg/kg bodyweight every 12 hrs. for 5-7 days. 2. The group t-PAHep (71 pts.) received 15 mg oft-PA in bolus followed by 50 mg in 30 min and 35 mg within the next 60 min; t-PA was followed by heparin 1000 u/hour for the next 48-72 hours. All the patients received aspirin. Three noninvasive CR criteria were used: 1. Rapid cesation of the chest pain. 2. Rapid decrease of the ST segment elevation by more than 50% from the initial value. 3. Rapid increase of the CK and CK-MB with a peak within the first 12 hrs. RESULTS: Two patients (2.85%) from the t-PAHep group had non-fatal stroke (one haemorrhagic, one ischemic). No other major haemoragical events were registered in both groups. During the thrombolytic infusion hypotension appeared more frequently in the SK0.75Enox group (31.4%) than in the t-PAHep one (8.5%) (p>0.0001) but without any consequence regarding the patients' outcome. The ratio of CR was 78.4% in the SK0.75Enox group and 70.4% in the t-PAHep one (p = 0.308). In-hospital reocclusion appeared in 4 pts. from the t-PAHep group (5.7%) but in none in the SK0.75Enox one. Six pts. (5.9%) from the SK0.75Enox group and 5 pts. from the t-PA one (7.04%) died within the first 30 days after the onset of STAMI (p = 0.993). CONCLUSIONS: The combination SK0.75Enox is at least as safe and efficacious as the t-PAHep one. PMID- 15526523 TI - Circulating osteoprotegerin and leptin levels in postmenopausal women with and without osteoporosis. AB - Osteoprotegerin (OPG) is a recently identified citokine with an important role in bone remodeling, that acts as a decoy receptor for RANKL; OPG was shown to be an important inhibitor of osteoclast differentiation and activation. Leptin influences bone metabolism by acting on differentiated osteoblasts, having an anabolic effect on bone. The relationship between circulating OPG levels and osteoporosis in postmenopausal women is controversial. Thus, one of the aims of our study was to investigate the relationships between OPG levels and biochemical markers of bone turnover and bone density in women with and without osteoporosis. We have investigated 135 postmenopausal women, including a group with osteoporosis (n=76, mean age 59+/-8 years) and a group with severe osteoporosis (n=31, mean age 64+/-8 years), using healthy postmenopausal women (n=28, mean age 48+/-9 years) as controls. The serum concentrations of OPG were determinated by ELISA. Serum estradiol was measured by Enzyme Immunoassay (EIA). The markers of bone formation and resorption were measured by standard methods. Leptin was measured by ELISA. Bone mineral density at lumbar spine and femoral neck was measured by dual energy x-ray absorptiometry (DEXA). There was a significant positive association between serum OPG levels and age (r=0.27; p<0.001), both in the postmenopausal women as a whole and in the cohort with osteoporosis. Circulating OPG levels were significantly higher in both osteoporotic groups (p<0.005 and p<0.01, respectively) than in the control group. There were no significant associations between serum OPG levels and bone density, bone markers and serum estradiol. Serum leptin levels were significantly associated with age (r=0.18, p<0.03), estradiol (r=0.2, p<0.05) and BMD (r=0.25, p<0.008); there was no significant relationship between leptin and bone turnover markers. We conclude that serum OPG levels increase with age, both in healthy and osteoporotic postmenopausal women. This could represent a possible protective mechanism against bone loss. Serum leptin levels also increase with age and are positively associated with estradiol and BMD and not significantly associated with bone turnover markers. PMID- 15526524 TI - Study of total homocyst(e)ine levels in type 2 diabetic patients with silent brain infarction. AB - Hyperhomocysteinemia is thought to have an important role in the pathogenesis of ischemic cerebral infarction. When associated with diabetes mellitus, it might worsen the neurologic course. The aim of the study was to clarify the relation between plasma homocysteine (Hcy) concentrations and silent brain infarction (SBI) in patients with type 2 diabetes mellitus. Total plasma Hcy levels were prospectively studied in 46 patients with type 2 diabetes and SBI (group I), mean age 56+/-5.4 years, as compared to 38 diabetic patients without SBI (group II) and with 31 controls (group III). Homocysteine concentrations were determined using a high-performance liquid chromatography assay. The results were compared using the Student's t test. The mean level of Hcy was 22.6+/-2.4 micromol/l in group I, 19.7+/-1.6 micromol/l in group II and 11.4+/-1.4 micromol/l in group III; between group I and group II p < or = 0.001. These data are consistent with increased Hcy levels in type 2 diabetic patients, contributing to the onset of SBI in some patients. The phenomenon should be considered in any future strategy for the therapy of hyperhomocyst(e)inemia (HHcy). PMID- 15526525 TI - Vasospastic angina caused by 5-fluorouracil. A case report. PMID- 15526526 TI - Systemic dermatitis and obstructive respiratory syndrome following occupational sensitization to trichloroethylene. AB - We present a derma-respiratory syndrome in a patient occupationally exposed to trichloroethylene (TCE). At the beginning of its industrial use trichloroethylene was considered harmless. But, in time it showed a high noxious capacity. It produces an important and various pathology, which evolves as acute or chronic disease. The case we present shows that trichloroethylene can induce cutaneous pathology that excels contact dermatitis. It also proves that trichloroethylene can produce systemic effects (obstructive respiratory syndrome). The particularity of the case is based on the succession of the events, first the cutaneous and then the respiratory effects. A long period of time was necessary for the installation of the symptoms (for cutaneous and bronchial sensitization to take place). The case presented is the proof that trichloroethylene's great toxicity cannot be doubted and that the clinical forms due to sensitization to trichloroethylene can be dramatic. PMID- 15526527 TI - Hypoplastic myelodysplastic syndrome transformed in acute myeloid leukemia after androgens and cyclosporin. A treatment. AB - The apparent contradiction between clonal expansion and marrow failure encountered in myelodysplastic syndromes (MDS) is more evident in hypocellular forms at presentation. Hypoplastic MDS (hMDS) appears to be a distinct clinicopathologic entity, accounting for about 15% from all MDS. The pathogeny is supposed to result from immunosupressive mechanisms and some observations on successful treatment with Cyclosporine A (CsA) are reported. The case of a young female patient diagnosed by bone marrow core biopsy with hMDS - refractory anemia (FAB and WHO classification) with normal karyotype and scarce CD34(+) cells by immunohistophenotyping is presented. She was treated with androgens followed by CsA for a few months and shortly after she developed an acute myeloid leukemia (M4) which responded to low-doses of daily oral melphalan. This is one of the first few reports on such an event during the immunosuppressive therapy in MDS and the possible explanations for this unusual evolution are discussed. PMID- 15526528 TI - Brain virus: a different kind of infection. PMID- 15526529 TI - Imaging of meningitis. PMID- 15526530 TI - Imaging of neurocysticercosis. PMID- 15526531 TI - Cerebral tuberculosis. PMID- 15526532 TI - Imaging viral and prion infections. PMID- 15526533 TI - Neurosarcoidosis--review of imaging findings. AB - Five to 25% of patients with systemic sarcoidosis also have CNS involvement. Patients with systemic disease may initially present with neurologic symptoms, and rarely, the sarcoidosis may be isolated to the CNS. A typical imaging feature is thickening and enhancement of the basilar leptomeninges of the brain. Other imaging findings, such as enhancing or nonenhancing parenchymal lesions, dural, and bone lesions may occur in the head and spine. Because there is a high rate of progression and recurrence following treatment, imaging follow-up is recommended in all patients. PMID- 15526534 TI - Fungal infections of the central nervous system. PMID- 15526535 TI - Case of the season: metastatic pituitary carcinoma mimicking meningioma. PMID- 15526536 TI - The effect of Simvastatin upon serum pseudocholinesterase and plasmatic factor VII. AB - The statins have a moderate lowering effect upon triglycerides, one hypothesis suggesting an inhibitory effect upon liver VLDL synthesis. Because VLDL synthesis is related to other hepatic protein synthesis we studied the effect of Simvastatin upon serum cholinesterase activity and factor VII activity, which are also synthetized at hepatic level. METHODS: 39 patients with ischaemic heart disease, proved by coronarography and moderate hypercholesterolaemia received for two months 20 mg Simvastatin (Zocor). The value of total Co, LDLCo, HDLCo, triglycerides, serum cholinesterase activity and factor VII activity were determined before and after treatment, by standard methods. RESULTS: Triglycerides were, together with total Co, significantly lowered during the two months treatment with Simvastatin, but the values of serum cholinesterase and factor VII activity were not significantly modified. This suggests that moderate doses of Simvastatin do not depress liver protein synthesis. CONCLUSION: Simvastatin does not lower the triglycerides level through depression of liver protein syntesis, other mechanisms being probably involved. PMID- 15526537 TI - Streptokinase and enoxaparin in the pre-hospital management of the ST-segment elevation acute myocardial infarction. AB - In a pilot study, safety and efficacy of an accelerated Streptokinase (SK) regimen (0.75 MU/10 min. repeated after 50 min. if no signs of coronary reperfusion were detected) in combination with enoxaparin was evaluated in the prehospital (preASENOX regimen) and in the in-hospital (in ASENOX regimen) phase of the ST segment elevation acute myocardial infarction (STAMI) and compared to the in-hospital standard SK plus Heparin (the StSK regimen). METHODS: A group of 262 consecutive patients (age 34-74 years) thrombolised within the first 6 hours after the onset of STAMI was divided in three subgroups according to the mentioned regimens: preASENOX (64 patients); in ASENOX (69 patients) and StSK (129 patients). Enoxaparin was administered i.v. 40 mg before the first dose of 0.75 MU of SK and 1 mg/kg every 12 hours for 5-7 days. Heparin was infused 1000 i.u./h 48-96 hrs. Three noninvasive reperfusion criteria were used: 1) Rapid cessation of the chest pain; 2) Rapid decreasing of the ST segment elevations by more than 50% from the initial value; 3) Rapid increasing of the CK and CK-MB with a peak within the first 12 hrs. RESULTS: The chest pain-thrombolysis time was 145 +/- 52 min in the preASENOX subgroup, significantly shorter as that in the ASENOX subgroup (172 +/- 73 min, p = 0.016) and the StSK one (168 +/- 80 min, p = 0.038). The ratios of the CR were 81.2%, 78.2% and 62.0%, respectively (preASENOX vs StSK p = 0.025; in ASENOX vs StSK p = 0.030). The inhospital mortalities were: 3.12% (preASENOX); 5.8% (inASENOX) and 10.8% (StSK)(non significant differences). Only one case of ischemic stroke was registered (in the inASENOX subgroup). Symptomatic hypotension appeared more frequent in the preASENOX (39.06%), and inASENOX (43.47%) subgroups as compared to the SSK one (20.15%). CONCLUSIONS: 1) The preASENOX and inASENOX regimens are safe, and lead toward a significant higher ratios of coronary reperfusion as compared to StSK regimen; 2) Our data suggest a very low mortality in patients treated with the preASENOX regimen. Further investigations (randomized studies) are needed for a definite conclusion. PMID- 15526538 TI - Cytokine patterns and pathogenicity in autoimmune diseases. AB - Cytokines are a large family of small proteins secreted by leukocytes and having an essential role in mediating the immune function. Many cytokines have multiple cellular sources and targets, as well as many natural inducers and inhibitors. These features and moreover the functional particularities of cytokines (autocrine and paracrine mode of action, overlapped activities, pleiotropic action, functioning as a complex regulatory network, and reciprocal down regulation of the Th1 and Th2 cytokine groups) hampered the investigation of cytokines' role in autoimmune diseases (ADs). Despite this, the experimental and clinical studies have firmly documented the implication of cytokines in major pathophysiological and pathogenetic mechanisms associated with both the acute (onset and/or recurrence) and the chronic stages of ADs. The enhanced production of cytokines during the acute phase of systemic lupus erythematosus, rheumatoid arthritis and vasculitis has pathogenic effects such as appearance of anti neutrophil cytoplasmic antibodies, initiation of vascular thrombosis and/or an increased production of autoantigens. The potent proinflammatory cytokines IL-1 and TNFalpha and also IL-8 and IFNalpha,gamma stimulate cells in different tissues to release harmful proteinases and reactive oxygen species, contributing to tissue damage. Some correlations have been found between serum cytokine levels and disease activity in certain systemic ADs. Studies investigating the cytokine pattern in ADs revealed the prevalence of the proinflammatory Th1-type cytokines in the target organ of patients with organ-specific ADs, such as multiple sclerosis and autoimmune thyroid disease, and heterogeneous cytokine profiles in patients with systemic ADs. A lot of factors, including the mechanism of tissue damage, the type of antigen-presenting cells and costimulatory molecules, and also the hormonal status, favor secretion of cytokines of Th1- or Th2-type in these patients. Beneficial effects have been obtained in patients with rheumatoid arthritis and Crohn disease by administering biotechnologically manufactured anti TNFalpha antibodies or TNFalpha receptors. The risks of these cytokine-targeting therapeutical interventions are also discussed. PMID- 15526539 TI - Serum cholinesterase activity correlates with serum insulin, C-peptide and free fatty acids levels in patients with type 2 diabetes. AB - When compared with values recorded in 14 control subjects, the 15 overweight patients with type 2 diabetes displayed significantly increased activities of serum alanineaminotransferase (172% of mean values in controls), gamma glutamyltransferase (253%) and cholinesterase (139%). A much wider dispersion of individual values for the two firstly mentioned enzymes was however noted so that their correlation with serum triglycerides levels were weaker (r = 0.373; p < 0.05 and r = 0.451; p < 0.05 respectively) than the same correlation obtained for serum cholinesterase (r = 0.760; p < 0.001). In two other studies including 28 controls and 30 diabetic patients serum cholinesterase was found to be significantly correlated with serum levels of insulin (r = 0.622; p < 0.001), C peptide (r = 0.652; p < 0.001) and free fatty acid (r = 0.821; p < 0.001). Circumstantial evidence is provided that insulin resistance and an increased flux of free fatty acids from adipose tissue to the liver would stimulate the hepatic synthesis of serum cholinesterase. PMID- 15526540 TI - Diabetic neuropathy--choices of treatment. AB - Diabetic neuropathy represents a heterogeneous pathology taking place during diabetes mellitus (DM) evolution, hiding possibilities for rapid evolution which threaten even the life of the diabetic person. As a result, the recognition as precocious as possible of the neuropathy-induced alterations and the ways of treating this complication are essential. Diabetic neuropathy is classified in peripheral and vegetative, each of them with numerous clinical forms. Nevertheless, its etiopathogeny remains not totally understood, many theories existing for explaining it, its development being probably the result of a mixture of pathogenic mechanisms. Although there are difficulties in appreciation of the neuropathy prevalence in diabetic people, it is generally known that this alteration is one of the most frequent and invalidant major complication that affects this population, treatment as precocious as possible being essential. alpha-lipoic acid, substance with antioxidant properties, plays a central role in the energetic metabolism, principally functioning as a coenzyme in multienzymatic mitochondrial complexes alpha-lipoic acid has complex actions on different levels of the human organism, having many indications in different diseases, including DM. The results of numerous clinical studies have confirmed its efficacy in treating diabetic neuropathy. The present study has proposed itself to investigate the long-term effect of the short-term i.v. treatment with Thiogamma 600 (alpha-lipoic acid) in patients having type I or 2 DM, proving neuropathy symptoms and signs reduction evaluated in 3 months time, without metabolic control of the disease. If this amelioration proves to be persistent in time, with or without continuing the medication, this treatment will represent the most evident and efficient present solution in neuropathy. PMID- 15526541 TI - Effects of Atorvastatin on some inflammatory parameters in severe primary hypercholesterolemia. AB - Recent publications have reanimated the point of view that there exist links between atherosclerosis--inflammation and hypercholesterolemia. The aim of our study was to investigate the possible influence of statins on some inflammatory parameters in persons with severe primary hypercholesterolemia (PHC). The effects of the HMG CoA reductase inhibitor--Atorvastatin--on serum lipids, apoproteins, C reactive protein (CRP), soluble Intercellular Adhesion Molecule (sICAM), lipid peroxides, antibodies to oxidized LDL (Ab oxLDL) and homocystein were evaluated in 21 persons (52.9 +/- 8.38 years old) with severe PHC, 12 of these having significant coronary-artery stenosis (diameter stenosis > or = 70%), in at least one major coronary artery branch. Ab oxLDL, sICAM, TBARS, CRP and homocystein were significantly increased (p < 0.05) in patients with coronary-artery stenosis. Following a 4 weeks hypolipemiant free baseline period, all persons were treated with Atorvastatin 40 mg once daily for 8 weeks. Atorvastatin 40 mg resulted in a reduction of LDL-C with 57.8% (baseline 259.6 +/- 71.39 mg%) p < 0.001, total Cholesterol with 44.08% (baseline 343.1 +/- 71.72 mg%) p < 0.001, Apo B with 50.6% (baseline 194.7 +/- 48.71 mg%) p < 0.001, TG with 12.02% (baseline 177.4 +/- 83.63 mg%) and HDL-C was increased with 6.84% (baseline 48.0 +/- 7.86 mg%). In coronary heart disease patients, Atorvastatin reduced homocystein concentrations with 19.41% (baseline 17.7 +/- 11.16 microM/l) (p < 0.01), and CRP with 21.9% (baseline 4.8 +/- 4.19 mg/l) p < 0.01 and TBARS with 52% (baseline 0.87 +/- 0.89 nM/ml) p < 0.001, but did not influence sICAM and Ab oxLDL. Thus atherogenic concentrations of LDL-C have to be closely modulated by minimal changes in LDL oxidative state. The effects of Atorvastatin on inflammatory parameters may crucially contribute to the clinical benefit of statins, independent of cholesterol lowering. Plaque stabilization may be a paradigm for antiinflammatory mechanism of action by this class of drugs. PMID- 15526542 TI - The analysis of structural lipid of the hair of patients with angina pectoris. AB - A study of 48 patients with angina pectoris carried out, in which the structural lipids of the hair were analyzed and correlated with the manifestations of angina and the potential incidence of dyslipidemia. It was noted that the excesses of hair lipids associated with deficiencies of some unsaturated fatty acids raised the frequency of angina attacks in the group studied by 2.7 times. The normalization of the lipid levels diminished the levels of angina. Data were obtained on the influence over a long time (weeks, months) of the hypolipidemic treatment. The analysis of the hair has been shown to be also useful in detecting latent dyslipidemia associated with angina pectoris. Also, through the analysis of hair, large scale epidemiologic studies can be carried out on the association between dyslipidemia and angina pectoris. PMID- 15526543 TI - The risk factors of fibrosis in nonalcoholic steatohepatitis. AB - Nonalcoholic steatohepatitis (NASH) can be a severe progressive liver disease leading to the development of cirrhosis. The aim of this study was to assess the risk factors of fibrosis in NASH. METHODS: The clinical, biochemical and histological features of 80 patients with NASH admitted at University Hospital from 1998 to 2000, were analyzed. The patients had no alcoholic, viral, autoimmune, drug induced or genetic liver disease. The fibrosis and necroinflammatory activity was associated with clinico-biochemical parameters and with oxidative stress. The clinical and biochemical parameters consist of: age, sex, body mass index (BMI), serum glucose, tryglycerides, alanine aminotransferase (ALT) and serum iron study (transferring saturation and ferritin). Liver iron overload was assessed by Pearls'staining and graded in four classes. The oxidative stress was evaluated by the levels of serum malon dyaldehide (MDA) and serum total gluthatione (GSI4). RESULTS: Septal fibrosis was present in 30 patients (27%) including cirrhosis in 4 patients (5%). Age > 45 years, B.M.I. >30 Kg/m2, serum tryglycerides >180 mg/dl, hyperglycemia > 220 mg/dl, serum ALT > 3N, increased hepatic iron and transferrin saturation percentage were independently associated with sepal fibrosis. Linear regression analysis showed that increased hepatic iron had the greatest association with the increase of lipid peroxidation (MDA > 250 nanomol/dl) and the decrease of serum gluthatione (< 40 micromol/dl). CONCLUSIONS: The clinical and biochemical parameters which consist of age, BMI, serum ALT, glucose and tryglycerides, Perls' grade and the serum index of oxidative stress (MDA and GSH) are independent risk factors for fibrosis in NASH raising therapeutic implications for the management of these patients. PMID- 15526544 TI - Chronic viral hepatitis, the treatment with spiruline for one month has no effect on the aminotransferases. AB - CONTEXT: Spirulina platensis is extracted from an alga and theoretically has many good effects on the majority of the organs. There is not any published clinical trial on humans. OBJECTIVE: The evaluation of the efficacy of spiruline in chronic viral liver disease. DESIGN: Double blind, randomised clinical trial. SETTING: Secondary care university hospital. PATIENTS: 24 patients with chronic viral liver disease, treated with spiruline or placebo for one month. OUTCOME MEASUREMENT: Aminotransferases diminution and the modification of a general state score self-evaluated by the patient on an analogic visual scale. RESULTS: A modification of the aminotransferases level in the detriment of the spiruline treated group has been found (p = 0.036 for ALAT, p = 0.017 for ASAT), and not at the level of the general state score (p = 0.30). CONCLUSION: Despite the little number of patients, significant results not favouring spiruline have been found. PMID- 15526545 TI - Clinical and hematological aspects of chronic myelomonocytic leukemia. Study on 20 cases from a single center from Romania. AB - Chronic myelomonocytic leukemia (CMML) has long been recognized as a disorder with both myelodysplastic and myeloproliferative characteristics, some patients showing clinical and morphological features resembling myelodysplastic syndrome (MDS) especially refractory anemia with excess of blasts (RAEB) with monocytosis, and others leukocytosis with neutrophilia, monocytosis and splenomegaly resembling myeloproliferative syndrome (MPS). The intrinsec differences determined at first the separation of CMML in two forms, one named "dysplastic", more similar with RAEB, and the other "proliferative", closer to chronic myeloid leukemia and then included by the recent WHO classification into a separate new created group--myelodysplastic diseases (MDD)/chronic myeloproliferative diseases (CMPD). The aim of this study was the analysis of some features of 20 cases of CMML, with emphasis on the differences between the two forms. The proliferative form of CMML differed from the dysplastic one by greater white blood cells and neutrophils counts (P < 0.001), a more important monocytosis in periferal blood (P = 0.07), and by the size and frequency of splenomegaly (P = 0.03). The sex and age of the patients, the frequency of the general symptoms, the frequency of the general symptoms and signs and that of infections, the hemoglobin and hematocrit values, the platelet counts, the percentage of myeloblasts and monocytes in bone marrow, the frequency of dysplastic traits, the percentage of reticulocytes and the modified Bournemouth prognostic index were not significantly different. These findings support the concept that CMML is a heterogeneous "overlap" syndrome between MDS and CMPD. PMID- 15526546 TI - Study of some markers of inflammation in atherothrombotic pathogenesis of acute ischemic stroke. AB - Recent researches focused on the study of the role of the inflammation in the atherothrombotic pathogenesis of the acute cerebral ischemia. The aim of the study was to identify some acute phase proteins with possible role in the pathogenesis of the ischemic stroke. Some acute phase proteins were prospectively investigated by standard methods in sera of 78 patients with ischemic stroke in the first admission day. There were two groups according to neurological deficit one month after the ischemic stroke: good outcome and poor outcome. In the second group mean value of C-reactive protein (CRP) was 0.122 +/- 0.06 g/l (p < 0.01), mean value of C3 was 2.61 +/- 0.36 g/l (p < 0.01), mean value of C4 was 0.73 +/- 0.07 g/l (p < 0.05), mean value of alpha 1-antitrypsin (AAT) was 4.9 +/- 0.46 g/l (p < 0.01), mean value of alpha 1-antichymotrypsin (ACT) was 0.33 +/- 0.04 g/l (p < 0.01), mean value of alpha 1-acid glycoprotein (AGA) was 1.12 +/- 0.15 g/l, (p < 0.05), mean value of fibrinogen was 2.6 +/- 0.22 g/l (p < 0.01), mean value of haptoglobin was 2.8 +/- 0.33 g/l, (p < 0.05), mean value of transferrin was 2.8 +/- 0.26 g/l (p < 0.05), mean value of ferritin was 238 +/- 22.42 microg/l (p < 0.001), mean value of fibronectin was 2.14 +/- 0.17 g/l (p < 0.05), mean value of ceruloplasmin was 1.23 +/- 0.24 g/l (p < 0.01). High significant values of ferritine and significant values of CRP, C3, AAT, ACT and fibrinogen were observed in patients with poor outcome. The presented data suggest that the studied markers are useful to appreciate the role of the inflammatory reaction in the atherothrombotic pathogenesis of the ischemic stroke. PMID- 15526547 TI - Oxidative aggression in atherosclerosis associated to degenerative psychoorganic disturbances. AB - We had in view the effect of the oxidative aggression as determinant factor in atherogenic process associated with degenerative psychoorganic disturbances. It was studied a group of old people distributed in two subgroups: a) 51 old people with atherosclerosis (AS) and b) 57 old people with atherosclerosis associated with degenerative psychoorganic disturbances determined by chronic ethylism. The results were compared to those of a 40 healthy adults group. Concomitantly, it were evaluated the supervened modifications in the antioxidant defense systems of the organism, by determining both of some nonenzymatic defense system components (reduced glutathione, ceruloplasmine and transferrin) and of some enzymatic defense system components (superoxide dismutase, catalase and glutathione peroxidase). The obtained results emphasize an important aggression exerted by the lipid peroxides against the old people organism, aggression that is amplified by the chronic ethylism with appearance of the degenerative psychoorganic disturbances. Generally, the antioxidant defense capacity of the old people organism is depressed, especially in the group with atherosclerosis associated with degenerative psychoorganic disturbances engendered by excessive and chronic alcohol intake. PMID- 15526548 TI - Higher order memories for objects encountered in different spatio-temporal contexts in mice: evidence for episodic memory. AB - The ability to build higher order multi-modal memories comprising information about the spatio-temporal context of events has been termed 'episodic memory'. Deficits in episodic memory are apparent in a number of neuropsychiatric diseases. Unfortunately, the development of animal models of episodic memory has made little progress. Towards the goal of such a model we devised an object exploration task for mice, providing evidence that rodents can associate object, spatial and temporal information. In our task the mice learned the temporal sequence by which identical objects were introduced into two different contexts. The 'what' component of an episodic memory was operationalized via physically distinct objects; the 'where' component through physically different contexts, and, most importantly, the 'when' component via the context-specific inverted sequence in which four objects were presented. Our results suggest that mice are able to recollect the inverted temporal sequence in which identical objects were introduced into two distinct environments. During two consecutive test trials mice showed an inverse context-specific exploration pattern regarding identical objects that were previously encountered with even frequencies. It seems that the contexts served as discriminative stimuli signaling which of the two sequences are decisive during the two test trials. PMID- 15526549 TI - Remembering emotional experiences: the contribution of valence and arousal. AB - Emotional experiences can be described by two factors: valence (how negative or positive) and arousal (how calming or exciting). Although both dimensions appear to influence memory, they may do so via distinct mechanisms. The amygdala likely plays a specific role in modulating memory for arousing experiences, whereas non amygdalar networks may be instrumental in enhancing memory for non-arousing positive or negative events. PMID- 15526550 TI - TMS induced plasticity in human cortex. AB - Repetitive transcranial magnetic stimulation (RTMS) is a novel, non-invasive and painless technique to stimulate the human brain with the intent to alter excitability or function of the stimulated cortex or its connections. This review focuses on RTMS induced changes in excitability, the potential mechanisms underlying these effects, and the usefulness of this knowledge to gain insight into mechanisms of other processes such as learning. PMID- 15526551 TI - Neuronal structure is altered by amyloid plaques. AB - During the course of Alzheimer's disease (AD), neurons undergo extensive remodeling, contributing to the loss of function observed in the disease. Many brain regions in patients with AD show changes in axonal and dendritic fields, dystrophic neurites, synapse loss, and neuron loss. Accumulation of amyloid-beta protein, a pathological hallmark of the disease, contributes to many of these alterations of neuronal structure. Areas of the brain displaying a high degree of plasticity are particularly vulnerable to degeneration in Alzheimer's disease. This article describes neuronal changes that occur in AD, reviews evidence that amyloid-beta contributes to these changes, and finally discusses the recovery of amyloid-induced changes in the brains of transgenic mice, lending hope to the idea that therapeutic strategies which reduce amyloid-beta production will lead to functional recovery in patients with AD. PMID- 15526552 TI - Neural mechanisms of versatile functions in primate anterior cingulate cortex. AB - The anterior cingulate cortex (ACC) is located on the medial wall of the cerebral hemisphere in humans and non-human primates, and has well developed intracortical and subcortical connections. Recent brain-imaging studies have suggested the possible involvement of the ACC in a variety of cognitive and motor-related functions. To clarify the cellular mechanisms underlying such higher-order functions in the ACC, neuronal activity in distinct areas of the ACC and its adjacent cingulate areas has been examined, through single unit recordings, in monkeys performing specific tasks. Each of the rostral (CMAr), dorsal (CMAd), and ventral (CMAv) cingulate motor areas basically participates in motor-related functions such as preparation and execution of movements. In particular, the CMAr appears to be involved in selection of appropriate motor responses as well as in planning of sequential movements. Furthermore, the CMAr and area 32 may participate in attentional functions which are necessary to select correct actions. These areas have also been implicated in detection of error actions and/or monitoring of action performance. Finally, a number of neurons in the ACC exhibit specific or modulated activity relevant to reward expectation. The primate ACC may play critical roles in performing appropriate actions with attention and in checking the performance to acquire rewards efficiently. PMID- 15526553 TI - The potential role of vascular endothelial growth factor in the central nervous system. AB - During the past decade, vascular endothelial growth factor (VEGF) has been widely investigated, and reported to have pleiotropic functions in the central nervous system (CNS) and its supporting physiological environment. VEGF is involved in not only such well-known functions as angiogenesis, accentuation of vessel permeability, and glial proliferation, but also more recently acknowledged functions such as neuroprotection and even neurogenesis itself. Most recently, the neurogenesis function has attracted much attention, and a number of research groups have taken up the challenge of elucidating this activity. In keeping with this trend, our knowledge of VEGF receptors has increased, and certain suggestions concerning the mechanisms of neuroprotection have come to light in the course of the ongoing work, though at times what the researchers had to work with was only a tiny percent of the signal transduction of VEGF. Together with flt-1 (VEGF receptor 1) and flk-1 (VEGF receptor 2), neuropilin (NP) is frequently described as being involved in the neuroprotective effects of VEGF. In this review, both the direct and indirect neuroprotective effects of VEGF, including various signaling pathways as well as the neurogenesis induced by this factor, are discussed in the context of the newly emerging insights into the biological mechanisms of VEGF and closely related, interacting molecules. PMID- 15526554 TI - [Economical impact of preterm delivery management based on fetal fibronectin results]. AB - BACKGROUND: Preterm labor continues to be the first cause (after congenital malformations) of morbidity and mortality during the perinatal period. One of the markers showing the highest sensitivity to predict preterm labor seems to be fetal fibronectin (fFN) determined in the cervicovaginal secretion. OBJECTIVES: To identify patients at risk of preterm labor based on the presence of fFN in cervicovaginal secretions and to analyze the cost-benefit of medical attention and hospital stay depending on the fFN results. MATERIAL AND METHODS: Four hundred and sixty two patients were admitted in a 6 month period to the Maternal Fetal Medicine Department. All of them had symptoms of preterm labor. Their gestational ages were between 24 and 34 weeks, the fFN analyses were taken from cervical vaginal secretion. One hundred and fifty eight were fFN positive and 304 were negative. Patients with positive fFN were hospitalized and received specific treatment for preterm labor. The patients with negative fFN were sent home with no medical treatment. In order to calculate the economic impact of the hospital's medical attention we considered the total cost in pesos which included patient's hospital care when admitted with preterm labor symptoms, hospital stay, and neonatal attention. The total costs were compared and analyzed in the two groups. RESULTS: The average cost generated for fFN positive and negative patients was 23,059 and 7,859 pesos, respectively. Approximately 15,200 pesos were saved per patient in this group. Multiplying this amount among patients with negative fFN (n=304), we would have saved 4,620,000 pesos in a 6 month period. Our established medical management did not affect negatively maternal-fetal well being. For statistical purposes of variables the Mann Whitney U, chi square and McNemar's tests were calculated. CONCLUSION: By determining fFN for the diagnosis of preterm labor, we obtained savings of 4,620,000 pesos in a 6 month period avoiding unnecessary treatments and hospital stay in patients with negative fFN. Determination of fFN in cervical vaginal secretion in patients with symptoms of preterm labor showed to have high sensitivity and specificity in predicting preterm labor between 24-34 weeks of gestation, permitting a more rational use of medical management and resources and avoiding unnecessary treatments. The treatment instituted on the basis of an opportune diagnosis in patients with positive fFN showed to prolong weeks of gestation. Although a persistence of morbidity and mortality of prematurity has been reported, these have diminished in comparison with studies previously done in the institution. PMID- 15526555 TI - [Factors related to the diagnosis of HELLP syndrome in patients with severe preeclampsia]. AB - BACKGROUND: Each year, around 50,000 women die from preeclampsia-eclampsia worldwide. Thus, hypertensive disorders during pregnancy are public health problems in both developed and developing countries. OBJECTIVE: To identify prognosis factors associated with HELLP syndrome in patients with severe preeclampsia. MATERIAL AND METHODS: A retrospective, observational, cross sectional, and analytical study was carried out. It included patients that suffered from severe preeclampsia, with and without HELLP syndrome. They were hospitalized at the Division of Obstetrics Dr. Manuel Gea Gonzalez General Hospital, from January 1st, 1995 to January 1st, 2000 (study group). Only clinical files of patients with severe preeclampsia, without convulsions, HELLP syndrome, or who had not died during the days spent at the hospital were included in the control group; within at least 72 subsequent hours to the pregnancy termination. The connection of HELLP syndrome with the following variables was assessed in the control group: gestational age, maternal age, infant formula, prenatal control, hypertensive disorder history, headache, tinnitus, phosphen, nausea, vomiting, epigastric pain, edema, hyperreflexia, blood pressure values, hepatic biometry, platelet count, blood chemistry with hepatic function. RESULTS: Right upper quadrant or epigastric pain was the most important independent prognosis factor. There were significant differences in the admission laboratory values between those with HELLP syndrome and those without acute complications of preeclampsia. CONCLUSIONS: Although the contribution of right upper quadrant or epigastric pain to the risk status of a pregnant patient is difficult to quantify, it can be used to assess whether the patient is at high risk for development of HELLP syndrome. PMID- 15526556 TI - [Pulmonary thromboembolism in obstetrics]. AB - BACKGROUND: Pulmonary embolism is a serious and sometimes mortal complication of pregnant and puerperal women. Pulmonary embolism diagnosis can be difficult. In Mexico, it causes between 2.5 and 16% of maternal mortality. OBJECTIVES: To estimate the incidence of pulmonary embolism, to identify most frequently risk factors that contribute to the presence of pulmonary embolism as an obstetrical complication and to determine clinical characteristics and specific diagnostic tests in hospitalized patients at the Hospital de Ginecoobstetricia, Centro Medico Nacional de Occidente, Instituto Mexicano del Seguro Social. MATERIAL AND METHODS: We conducted a cross-sectional study from January 1st 1997 to December 31st, 2002. We identified 30 patients with pulmonary embolism confirmed by ventilation-perfusion scan. RESULTS: Incidence of 4.7 cases per 10,000 pregnancies was found. Most frequently, risk factors were varicose veins in lower extremities (0.045), previous thromboembolic event (0.030) and pre eclampsia/eclampsia (0.05). Cesarean section was present in 85% of the cases with pulmonary embolism during puerperium. The most common clinical findings were: dyspnea (100%), chest pain (63%), tachycardia (93%) and tachypnea (93%). Sinus tachycardia (93%) and S1 Q3 T3 were the electrocardiogram findings most commonly observed. 83% of the patients showed abnormalities in chest radiography. 100% presented altered blood values. Mortality rate was of 6.6%. CONCLUSION: Diagnosis of pulmonary embolism should be based on risk factors and clinical findings. Basic laboratory and scan are essential. Early diagnosis and treatment significantly reduce mortality rates. PMID- 15526557 TI - [A study of two different routes of administration of micronized progesterone in assisted reproduction]. AB - BACKGROUND: Progesterone is almost completely absorbed after administration by oral route when its preparation is micronized, but, due to the important metabolic inactivation during the first hepatic pass, bioavailability of oral progesterone is notably poor reaching values lesser than 10%. OBJECTIVE: To compare two different routes of administration of progesterone for luteal phase support in artificial insemination protocols. MATERIAL AND METHODS: 24 patients were randomly allocated in the 2 arms of the study, 200 mg/daily, vaginally or p.o. Plasma levels of progesterone were analyzed and ultrasound was performed to compare endometrium pattern and thickness. RESULTS: No differences were found in terms of indication, semen parameters, number of preovulatory follicles and progesterone serum levels. Endometrial pattern and thickness were clinical and statistically better in group I as the pregnancy rate. DISCUSSION: Taking in account our results we can argue that vaginal progesterone improve endometrial features favoring implantation and pregnancy rates without modifying progesterone serum levels. PMID- 15526558 TI - [Treatment of syphilis during pregnancy. 1949]. PMID- 15526559 TI - [Specific recommendations to improve medical care in the prenatal diagnosis]. PMID- 15526560 TI - Have GPs caused an A&E meltdown? PMID- 15526561 TI - A multi-ethnic approach to care is vital. PMID- 15526562 TI - The overseas nurse experience. PMID- 15526563 TI - Dreamcatchers: supporting bereaved children. Interview by Nic Paton. PMID- 15526564 TI - Challenging diabetes myths. Interview by Janis Smy. PMID- 15526566 TI - What you need to know about ... Meniere's disease. PMID- 15526565 TI - The NSF for children, young people and maternity services. AB - The new National Service Framework for Children, Young People and Maternity Services (Department of Health, 2004) draws together current recommendations for best practice. This article highlights the points most relevant to nurses from this 10-year plan of practice improvement. PMID- 15526567 TI - Using a clinical decision-making support tool to enhance practice. AB - As nursing practice expands nurses need easy access to reliable cohesive guidance. PRODIGY is a support tool for clinical decision-making which is approved by the Department of Health. It is a learning resource that provides information on about 170 illnesses and covers condition management, prescribing options and patient information leaflets. This article uses a fictional case study to illustrate how nurses can make use of PRODIGY guidance to provide holistic, patient-centred care. PMID- 15526568 TI - How the media has affected the discharge of mentally ill patients. PMID- 15526569 TI - A nurse-led ADHD service for children and adolescents. AB - Evidence-based nursing guidelines were developed on the management and treatment of attention deficit hyperactivity disorder (ADHD) in Forth Valley. After an audit revealed that adherence to the guidelines was generally good, except in terms of providing individualised school-based programmes for children with ADHD, a service was developed to work closely with schools and families to improve this area of care. PMID- 15526570 TI - Understanding the experience of training for overseas nurses. AB - AIM: To explore the perceptions of overseas nurses during their induction programme. SAMPLE: A pilot cohort of 20 overseas nurses. METHOD: A qualitative research approach was used. RESULTS: The key themes were: communication issues, culture, role definition and feelings of self-worth, which are interrelated and suggest how the experience has influenced each nurse's professional development and ultimate achievement of 'competence' as a registered nurse able to practise in the UK. CONCLUSION: These findings confirm there is a need for greater understanding of the 'adjustment process' and integration into the workforce. Recommendations are made that for future projects to succeed, comprehensive support frameworks are required to fully support both the overseas nurse and the organisation as a whole. PMID- 15526571 TI - Adapting to the challenges of a different health care system. PMID- 15526572 TI - [Back to the usual. We find what we left]. PMID- 15526573 TI - [Mobbing in nursing. A pilot study]. AB - While planning to help shed light on the phenomenon of mobbing in the work place and to develop an instrument by which to measure it, the authors carried out a pilot study in which 160 persons from varying professional classes and autonomous communities in Spain participated, 65 of whom were nurses whose ages lie between 20 and 48 years, with a medium age of 33.98. By means of a factorial analysis, the authors discovered that the most common mobbing practices are grouped in a set of eight factors; these eight factors cover 74.17% of all the various factors. The two most significant factors refer to behaviors which can be considered to be personal humiliation and professional discredit. The most common mobbing practice, according to the overall findings of this study group, consists in providing contradictory information to the victim (19.4% once or more times per week), while in nursing, this practice consists of exposing the victim to criticism by the group (50%). PMID- 15526574 TI - [Do patients and their families expect what we really offer them?]. AB - Frequently we evaluate user satisfaction for the treatment procedures provided (quality perceived), but we do not study the prior expectations (quality desired). Our objective is to discover what these patient and their family expectations are about treatment procedures and nurses' role so that we can make improvements and contribute to maximizing the quality of treatment provided. The aspects which cause greatest concern so that a patient's hospital stay is comfortable are the interpersonal relationships with family members, room mate, and professionals. Furthermore, the authors evaluate the treatment with nurses as a primary aspect in quality treatment. PMID- 15526575 TI - [Phlebitis in peripheral catheters (I). Incidence and risk factors]. AB - The type of pathology and the current therapeutic tendencies justify that endovenous therapy is the most frequently selected one. Although it has multiple advantages, it bears with it a series of infectious type complications (bacterial infections, phlebitis and thrombophlebitis). Even though bacterial infections in peripheral intravascular vessels have a low occurrence rate, which some authors list as between 1 and 2%, in central intravascular vessels, their occurrence is usually higher. Phlebitis is one of the complications most frequently associated with the use of peripheral intravascular catheters. PMID- 15526576 TI - [Phlebitis in peripheral catheters (II). A study]. AB - The study group consisted of 121 patients, 37 or 30.5% women and 84 or 69.4% men, checked into the Internal Medicine and Infectious Diseases Hospitalization Unit who bore a total of 215 peripheral venous catheters. The authors of this study detected a high level occurrence of phlebitis in patients using peripheral venous catheters, with a low average duration time, during the period in which the authors carried out their study. PMID- 15526577 TI - [How to reduce the incidence of traumatic phlebitis]. AB - The authors carried out an analytic study on 2572 patients to reduce the occurrence of phlebitis having a traumatic cause. The authors conclude that the use of extensions after three way valves, or infusion systems and a venous catheter, considerably reduces the risk of traumatic phlebitis occurring. Furthermore, their use reduces 1. the bother patients incur, 2. the amount of work nursing professionals have derived by the appearance of traumatic phlebitis and 3. the cost of health care. PMID- 15526578 TI - [Phlebitis in children]. AB - The authors' objectives are to determine the occurrence of phlebitis in patients hospitalized in the General Pediatrics Unit and to define the possible factors associated to its development. The authors carried out a prospective study in the "Sant Joan de Deu" Hospital in Barcelona during the month of July and from the 15th of September until the 15th of October in 2002. A one variant and two variant analysis using the chi squared method as the basis for estimating which associations took place. There were 153 patients admitted having 201 peripheral veins canalized; occurrence of phlebitis was 8.4%; a venous cord (53%) was the most frequent clinical sign. In cases of phlebitis, the back of a patient's hands (64.7%) was the main spot to make an insertion using a 22G "Vasocan" catheter (70.6%). Phlebitis appeared 2 days after insertion; the high number of antibiotics administered increased the risk of phlebitis (p>0.05). CONCLUSION: Phlebitis is a frequent problem in pediatrics patients having endo-venous treatment, susceptible to prevention by means of the implementation of control measures and a program of continuous vigilance. PMID- 15526579 TI - [Aneurysm in the aorta. Diagnostic confirmation by means of a helicoidal CAT scan]. AB - A helicoidal radiological CAT Scan, which produces reconstructions in varying planes and makes use of an injected contrasting liquid, confirms a diagnosis of an abdominal aorta aneurysm in the shortest time possible, whether this aneurysm is broken or dried, in order to facilitate a patient's treatment to the highest degree. PMID- 15526580 TI - [Prevention of pressure ulcers in heels]. AB - Heels are, for all assistance levels, one of the most frequent locations for the development of pressure ulcers (PU). In this study we deal to investigate in order to determine in patients at risk in an Internal Medicine Unit, the PU incidence on heels, after applying a specific prevention protocol. This protocol particularly designed for pressure ulcers on heels included a combined application of special hydrocellular dressings specially shaped for heels (Allevyn Heel), hyper-oxygenated fatty acids (Mepentol) and special surfaces for pressure management (Aerocare); afterwards, we attempted a comparison of our results with those from previous similar studies. We designed a prospective study which lasted from May 1-2002 until June 30-2003, with a sample of 100 patients without PU included in the study when admitted to the unit. The cumulated incidence established for PU in heels is a 4% which means an incidence rate of 2.06 PU in heels per 1000 persons/day. After observing the results we may affirm that applying the protocol is, under a clinical point of view, as effective as other measures used in previous studies. If we focus on the cost-benefit, the protocol studied represents an option with an excellent cost-efficiency relationship. PMID- 15526581 TI - [Following breast cancer surgery, which treatments are necessary?]. AB - This essay received first prize in the Third Nursing Conferences organized by the Sanitary Consortium of the Alt Penedes County in Catalonia. In a very clear, simple way, this essay deals with those questions which most concern women who have undergone an axillary extirpation. The authors describe their everyday experiences in health education for women who have undergone this surgery. PMID- 15526582 TI - [Preventing childhood accidents (and III)]. PMID- 15526583 TI - [Unusual chromosome translocation 46, XX, t(1;16) (p13;p13) as a possible cause of retarded development of carrier]. AB - Unbalanced changes of chromosomes with loss or extra genetic material often cause changing phenotype. The main characteristics unbalanced karyotype involves mental retardation and numbers of anomalies, especially if the change involves autosoms. In this paper child with retard development and de novo translocation between chromosome 1 and chromosome 16 was described. Cytogenetics survey was carried out to establishing possible cause in relation with retard development and it involved standard cytogenetics method and method of differential staining (G banding) that enables identification of certain structural rearrangement of chromosomes. Chromosome aberration was translocation of part of p arm of chromosome 1 on p arm of chromosome 16, that is, it presents abnormal karyotype 46, XX, t(1;16)(p13;p13) that might be one of the cause of retard development of child. PMID- 15526584 TI - Serum angiotensin converting enzyme in patients with psoriasis. AB - Serum angiotensin-converting enzyme (ACE) was measured in 60 patients with psoriasis and in 16 healthy individuals. According to clinical forms ofpsoriasis, the patients were further divided into three groups: psoriasis with solitary lesions (n=20), psoriasis with multiple disseminated lesions (n=20) and erythrodermic psoriasis (n=20). The serum ACE activity was determined before and after therapy, by the spectrophotometric method using hippuryl-l-histidyl-l leucine as a substrate. Before therapy, serum ACE activity was significantly increased in patients with psoriasis (47.20 +/- 2.06 U/L) in comparison to healthy individuals (28.33 +/- 1.32 U/L). The greatest increase in serum ACE activity was observed in patients with multiple disseminated lesions (78%), followed by those with solitary psoriatic lesions (76%) and erythrodermic psoriasis (31%). After therapy, serum ACE activity was significantly decreased in all clinical forms of the disease. In conclusion, the determination of serum ACE activity may be helpful in the diagnosis of psoriasis and one of the discriminators to assess the effects of used therapy. PMID- 15526585 TI - [Genotoxicity evaluation of paracetamol applying Allium test]. AB - Paracetamol genotoxic potential was evaluated among different concentrations, applying Allium test on Allium cepa. Total number of roots, number of dark roots, the length of the longest root, the average root, and the shortest root were determined. Statistically significant differences among total number of roots (p > 0.05) was observed at concentrations of 50 microg/ml and 100 microg/ml, and highly statistically significant differences at concentrations of 5 microg/ml and 25 microg/ml, while at the highest concentration (400 microg/ml) was observed statistically significant higher number of roots in comparation to all other concentrations of paracetamol and control group. The results of research suggest the concentrations of 5 microg/ml, 25 microg/ml and 400 microg/ml for further evaluation of paracetamol mutagenic potential. PMID- 15526586 TI - [Acute physical workload and glucose serum concentration in subjects with different level of physical activity]. AB - The aim of this study was to estimate glucose serum concentration during acute workload in the subjects with different level of physical activity. The study involved a group of healthy males, average 21 years of age, which, based on the level of their physical activity, were divided into two groups: trained and untrained subjects. Acute work load of 30 minutes duration was carried out on byciclergometer following the protocol of individually adjusted constant work load. The glucose serum concentration was measured in the rest and during exercise (6., 12. 18., 24. and 30. minutes). The standard glucose oxidase method was used to determine serum glucose concentration. The serum glucose concentration was in the mean refferal values in the rest period, and during exercise in both examined groups. In the trained group of subjects there was no statistically significant changes in glucose serum concentration during acute work load. In the untrained group of subjects we established decrease in glucose serum concentration during exercise (at 18th., 24th. and 30th) compared to the concentration at rest period. Level of physical activity does not cause significant changes in the glucose serum concentration during acute work load of moderate intensity. Trained subjects have better ability of serum glucose concentration maintenance during exercise compared with sedentary subjects. PMID- 15526587 TI - [Comparison of gastrointestinal toxicity in patients with advanced cervical carcinoma treated with concomitant chemotherapy and radiotherapy versus radiotherapy alone]. AB - BACKGROUND: Cervical carcinoma is the second most often malignant tumor of female in Bosnia and Hercegovina. About 70% of patients in the time of diagnosis have advanced disease, when prognosis and results of treatment are poor. Improvement of survival and local control of disease could be possible with combined chemo radiotherapy treatment, expecting higher gastrointestinal toxicity. AIM: Comparation of gastrointestinal toxicity in patients with advanced cervical carcinoma treated with concomitant chemo-radiotherapy versus radiotherapy alone. PATIENTS AND METHODS: From 1997-1999, we performed prospective, randomised study, and 80 patients were divided in two groups: Group A--40 patients treated with concomitant chemo-radiotherapy versus, Group B--40 patients treated with radiotherapy alone. Gastrointestinal toxicity was evaluated before enrollement, during the therapy, immediately after the treatment, 3 and 6 months, according to the CTC toxicity criteria. RESULTS: We performed statistical analysis using H2 test, and we found no significant differencies in mild and severe gastrointestinal toxicity among groups, while in the Grade 1 toxicity we found statistical differencies in nausea and vomitus, and no differencies in diarrhoa. PMID- 15526588 TI - [Acute polyradiculoneuritis--clinical course and outcome during the war and postwar period]. AB - The acute polyradiculoneuritis represent acute inflamatory polyneuropathy with unknown cause, frequently induced by prior virus and sometimes bacteria infection. The purpose of this paper is to establish particularities of the clinical course and results of the disease during the war and the post-war time. Within the retrospective study we performed analysis of all patients suffering from the acute polyradiculoneuritis between January 1992 and December 2003 year who during the mentioned period where treated at the Department of Neurology in Sarajevo. In addition to the anamnysis and the clinical examination, diagnostic criteria where laboratory parameters, EKG, cardiological finding, cerebrospinal liquor (LP) and the EMG finding. PMID- 15526589 TI - [Morphological characteristics of the phyllodes tumours of the breast]. AB - Tumor phyllodes arise from stroma component of the terminal ductulo-lobular unit (TDLU). Stromal and epithelial proliferation are present in the same time. The stromal component grow th more intensive and dominante over the epithelial component. Clinically, macroscopically and microscopically tumor phyllode look like fibroadenoma which is more common lesion than tumor phyllodes. The differentiation between these two lession is important because the fibroadenoma almost never reccur bur the reccurence is ofthen in tumor phyllodes especially in incomplete excision. The fibroadenoma growth is never over 3 cm, but the growth of tumor phyllodes is sometimes over the 20 cm. The clinica differentiation in smaller lesions is very hard, macroscopical differentiation is a litle better, but the final diagnosis is microscopical. Biological behavior is not in the correlation with histological picture. Histologically, we can differentiate benign, borderline and malignant tumor phyllodes. PMID- 15526590 TI - [Clinical aspects and haemodynamic parameters for monitoring patients with acute myocardial infarct (AIM)]. AB - In acute myocardial infarct (AIM) incomes to a local disturbances in contractility: hypokinesia, akinesia, diskinesia, asynchronia. With repeated AIM esspecially in high oclussive coronary disease incomes to diffuse disturbance in contractility of myocard, with disorder in systolic and diasystolic function of heart. The aim is to point on significance of clinical and haemodynamic parameters in revealing heart weakness in AIM. By retrograded analysis in our study was embraced 86 patients deseased from AIM, average age of 58.42 years. Front wall infarct had 60,46%, backwall infarct had 39.54% of deseased patients. Patients were monitored clinically, electrocardiographically and haemodynamically with microcatetherisation of the right heart by Schwan-Ganz threeluminal thermodilutional catether with regulation of heart minute volumen. By haemodynamic values they were splitted in four groups. In clinical study, silent heart tones had 33%, systolic murmurs on ictus 19%, gallop rhythm 26% and lung failure 22% of patients. The most reliable clinical sign for heart weakness were low heart tones and gallop rhythm as well as lung failure. By haemodynamic values aquired by microcatetherisation of right heart with regulation of minute heart volumen, deseased with AIM were 15% in first group, 28% in second group, 3% in third group and 54% in fourth group of patients. PMID- 15526591 TI - [Effects of iron therapy on motor and mental development of infants and small children suffering from iron deficiency anaemia]. AB - The aim of this paper is to establish the effect of iron therapy on mental and motor development in children suffering from iron deficiency anaemia by Bayley Scales of Infant Development (BSID). PATIENTS AND METHODS: In prospective study were enroled 90 anemic children with Hb < 110 g/l and 30 healthy children with Hb > 110 g/l, all of them 6-24 months of age. RESULTS: Indexes of mental development (MDI) before and 3 months after iron therapy for group of patients with severe and mild form of anemia (Hb < 95) did not show statisticaly significant difference (p > 0.05) before and after three months after iron therapy. Before iron therapy there was statisticaly significant difference (p < 0.001) between group with Hb < 95 g/l and group with Hb 95-110 g/l, and also statisticaly significant difference (p < 0.001) between anemic groups and control group. Indexes of psychomotor development (PDI) before and three months after iron therapy were similar to results MDI. CONCLUSION: level of Hb < or = 95 g/l had great importance for results BSID test in infants and small chilren up to age of two years. PMID- 15526592 TI - [Transdermal drug delivery systems]. AB - Transdermal drug delivery systems are pharmaceutical preparations, intended to be applied to the unbroken skin in order to deliver the active ingredient(s) to the systemic circulation after passing through the skin barrier. Transdermal patches are formulated in different ways, but normally they consist of release liner, adhesive layer and backing layer. Active ingredient(s) can be incorporated in reservoir, matrix, adhesive, membrane-matrix and microreservoir. The releasing rate of active substances from patches is controlled by membrane, matrix and adhesive. Only several transdermal systems are available on the market today, because for the penetration of most substances skin represents a strong barrier. With transdermal systems it is possible to obtain a controllable and sustained plasma levels, to minimaze the risk of undesirable side effects and to avoid the hepatic first- pass metabolism. These devices are easy to apply and also to remove from the skin. Of a great pharmaceutical interest are also socalled cosmetic patches. They are applied with aim of cleaning and protecting the skin. PMID- 15526593 TI - [Intracranial aneurysms treated by means Guglielmi detachable coil]. AB - AIM: The endovascular treatment of an intracranial aneurysm using the Guglielmi detachable coil (GDC) becomes more and more treatment of choice which is based on aneurysm configuration, aneurysm location, the patient's medical and neurological condition and age, available surgical and interventional abilities at the treatmant center, and patient preference. METHODS: Out of 73 patients with the intracranial aneurysm, 9 were treated by the endosaccular embolization using the Guglielmi detachable coil. Subarachnoid hemorrhage (SAH) occured in 8 patients and a spacio-compressive effect caused epilepsy in one patient. Three patients with multiple aneurysms were treated by the combination of microsurgical clipping and endosaccular embolization. The outcome was obtained after 6-12 months on the basis of Glasgow outcome scale. RESULTS: Out of five patients treated in acute phase of severe subarachnoid hemorrhage (Hunt-Hess grade 4 and 5), one patient died for an inicial hemorrhage, one had a severe disability, and three patients had a good recovery or moderate disability. Two patients with the multiple aneurysms returned to the same quality of life as before the SAH, and one patient had moderate disability. CONCLUSION: Microsurgical clipping of the neck of aneurysm is an optimal way of treatment for the most of ruptured intracranial aneurysms. Endosaccular embolization becomes more and more the treatment of choice in selected cases. PMID- 15526594 TI - [Trombosis of the middle cerebral artery as the cause of cerebrovascular insult (CVI) and recognition of the etiologic factors for CVI]. AB - In the paper is shown the case of the patient of the CVI life, age of 40 years. It was about the vascular lesion left of the temperoparienal MRA shew the amputation of amputation of the temporal branches art. cerebri medii from the left side. The patient 6 years before she suffered of CVI infract myocard after which she recovered well. After the performed neurologic and physiatric treatment it came to the strength of the rude motor strength with the normalization of the speech. PMID- 15526595 TI - [Importance of an interpersonal approach and object relation theories for the organization of treatment at the psychiatric hospital]. AB - The importance of interpersonal relationships for the organization of the psychiatric hospital as a therapeutic setting is presented through the theoretical framework of object relations and milieu therapy. From the point of view of psychodynamic approach the majority of hospitalized patients show the pathology of self and object relations. So, the familiarity with these theories for the optimal planning of treatment of these patients is of a particular importance. The concepts of following authors are discussed in this paper: Winnicot, Kohut, Mahler, and Klein, as well as a concept of projective identification and milieu therapy. PMID- 15526596 TI - [Urinary infection in patients with the condition of paraplegia]. AB - The data were retrospectivelly analyzed conserning the frequency of uroinnfect in person with the paraplegia condition at the institute for physiatry and rehabilitation--the Center for paraplegia of the Clinical center of University in Sarajevo. The analysis was involved 71 (10%) of the patients chosen by the method of the casual choice. Males 57 (80.28%), females 14 (19.71%) of the examimnees. Urolitiazation was found in 22 (30.98%), without urolitization was 49 (69.01%) of patients. The sterile urinoculture were found in only 4 (5.63%) of the patients. The greatest number of the patients 34 (47.88%) he had three and more bacteria in urinoculture. Per two bacterias had 18 (25.35%), with the a one bacteria was 15 (21.12%) of patients. That the degree of the education and enabling of the activities of every day life (AEL) has no essential influence on the seizures of uroinfect shows the result of the research of Barther index where we found 59 (83.09%) of the examinees independent in movement. What are social consequences of this condition when the complications by uroinfect and urocaliculosys can be seen by the hospitalization number at the Institute for the rehabilitation where 29 (40.84%) of the examinees was hospitalized more than three times, 12 (16.90%) three times, 16 (22.90%) and only 14 (19.71%) once. It is considered that the persons with the paraplegia condition if they have no the acute becoming works should perform at least the essential control examinations every six months. PMID- 15526598 TI - [The role of corticosteroids in the lungs' maturation in expected premature birth]. AB - Cortikosteroids, a hormons of adrenal glands have a influence to almost all organic sistems because of their antiinflamatory and immunosupressive effects. They are used in perinatology to accelerate lungs maturation in expected premature labour. The most frequent used corticosteroids are bethamethason and dexamethason, with useful effects: acceleration of growth of fetal lungs by ripening pneumocites type II and supression of creating secundary septs; by protection of brain from intraventricular haemorrhage and periventricular leucomalation; decreasing rate of rethinopathy and enterocolitis and ductus artheriosus persistens. By this way the mortality rate ofpreterm newborns is decreased in 60% of cases. Expected harmful effects of antenatal using of corticosteroids are: high blood pressure of newborns in first 24 hours after delivery, the possible oedema of lungs, in simultaneous using of corticosteroids and bethamimetic treatment. It is not recommended to repeat treatment by corticosteroids in same pregnancy because of hormonal influence to fetal growth and neurological development, till clear benefit is not proved. PMID- 15526597 TI - [Evaluation of functional treatment of the acute Achilles tendon rupture according to the Thermann score scale (ten years experience)]. AB - INTRODUCTION: The injury of Achilles tendon most frequently occurs as a result of overburdening of the tendon, particularly in sportsmen (runners and jumpers) even though they are not rare in the rest of the population. Biomechanical distrubances on the burdening tendon, or its continuous burdening, result in the degenerative changes in the form of pathological tenosynovial adhesions in and around the tendon that precedes the tendon rupture. In the recent years conservative functional treatment increasingly became the method of choice due to final results of the treatment. Basic principle of this method of treatment is to establish disrupted balance between the synergist and the antagonist of the lower leg, which was caused by rupture. This helps to reduce the kinetics of the ankle and the knee joint, and to avoid long lasting classical immobilisation which has found its substitution in a vario stable shoe, which allows possibility of flexibile arthrodesis of ankle with usage of insole. MATERIALS AND METHODS: The conservative functional treatment of the acute Achilles tendon rupture has been carried out on 19 patients during the period 1993-1998, at the University of Sarajevo, Clinical Center. Sex structure was 17 males and 2 female patients. Average age of the patients was 27,1 (working population). The study was shortterm with first results obtained after 12 weeks. Etiologically, in the mechanisms of the injury there were 15 spontaneous ruptures from jumps and 3 from in landing after a jump, and in a direct trauma there was one rupture reported. Two of these were sport injuries. Clinical evaluation has been done according to the Thermann score scale. Ultrasound diagnostics was done immediately after the injury, followed by control examinations in the 4th, the 8th, and the 12th week. If the vario stable boot is missing, the modified program can be applied. RESULTS: After 12 weeks, the Thompson, the Simmond and the Matles signs were evaluated and the results were negative. No patient reported suffering pain. The muscular atrophy of the lower leg up to 2 cm was reported in six patients, and up 3 cm in 3 patients, while other patients had tolerable 0.5 to 1 cm atrophy. There were no reports of thromboembolism. The largest number of ruptures was verified on the crossing between the middle and the lower third by ultrasound examination (8). Complete ruptures were reported in 14 examinees, partial in 5. There were no reports of desinsertions and distensions. In the 12th week we followed the ultrasound adaptation, i.e. diastases of tendon ends in neutral position (0(0)) and functional with 20(0) of plantar flexion. There were 16 examinees without functional deficit, one with the reduced plantar flexion od 5(0), 1 with the reduced dorsal flexion od 5(0), and also one patient with 10(0) reduced dorsal flexion. After ten years (2003-2004) we reevaluated six patients according to the Thermann scor scale. Functional examinations each patients have been excellent. CONCLUSIONS: All patients returned to their daily working activities. We did not record a single case with complicatoions. With regard to other methods of treatment the functional treatment is apparently without an alternative, but at the same time requires methods that seek full engagement of a surgeon, a physiatrist and the radiologist. In order to avoid compromising and make the treatment successful, it is necessary to keep up with the implementation protocol, to have available adequate corrective shoes or other means, and to possess certain level of clinical experience. PMID- 15526599 TI - [Establishment of the first medical faculty in Bosnia and Herzegovina]. AB - After establishment of the medical faculties in Zagreb (1917 year), Ljubljana (1919 year) and Belgrade (1921 gear), by the decision of the minister of the National education of the Independent State Croatia from 31.3.1944. year, there were establishment the medical faculties in Sarajevo and Split the same year. The Medical faculty in Sarajevo, as the faculty of the University in Zagreb, officially began with the work 22.11.1944. year by the opening which made the then dean of the Medical faculty in Zagreb professor (doctor) Ante Sercer. For the dean of the Medical faculty in Sarajevo was nominated professor Stanko Sielski, the renown bosnian-herzegovinian physician, the former head of the district bacteriological station in Tuzla and Banja Luka. The access lectures held professor (doctor) Mile Budak, professor (doctor) Ljudevit Thaller, professor (doctor) Ibrahim Ruzdic. The first generation of the students of the school 1944/1945 year numbered about 165 students. The teaching was organized in the complex of the building of the present residance Conac in Sarajevo, and the practical instruction was performed on the basis of the contract with the then General hospital in Sarajevo. The teachers of the faculty were the eminent professors of the Medical faculty at the University in Zagreb. From all the known person, the fact about the so called NDH (IS Croatia) to the medical faculty, for the long time were hidden from the publicity, so that about them was not spoken, till the back several years, either the momentary alive students of the generation. In this article is stated the factographia connected for the establishment and functionioning of the mentioned first medical faculty in Bosnia and Herzegovina. PMID- 15526601 TI - [Anaplasmosis in a Swiss dairy farm: an epidemiological outbreak investigation]. AB - In summer of 2002, a case of severe clinical bovine anaplasmosis caused great losses in a dairy farm of an animal trader in Grisons. This article outlines the general approach of an outbreak investigation considering the case of anaplasmosis as an example. The goals of such investigations are to identify and eliminate the source of a disease outbreak in order to avoid additional cases. In addition, recommendations should be developed for preventing or limiting the magnitude of similar outbreaks in the future. In the outbreak presented, the causative agents were probably brought into the dairy farm by animal trade. Due to the large herd size, a missing quarantine for new animals and the coinfection with several pathogens, this case led to a high number of fatalities. The investigations of this outbreak demonstrated the importance of an universal and consistent identification of individual animals for the reconstruction of their movements. The veterinary practitioner should be reminded to act cautiously when facing strange clinical cases and to also consider "exotic diseases" as a possible cause. PMID- 15526602 TI - Porcine circovirus as a possible cause of postweaning wasting in pigs in Switzerland. AB - Postweaning wasting is a major worldwide problem in pig production, particularly with respect to the disease termed postweaning multisystemic wasting syndrome (PMWS). In addition to wasting, PMWS symptoms include respiratory distress, diarrhoea, pallor and occasional cases of jaundice. The causative agent is porcine circovirus type 2 (PCV-2). The objective of the present study was to determine the significance of PMWS and similar conditions in Switzerland. A total of 72 weaned piglets from 26 farms showing wasting were examined for the presence of PCV-2 by immunohistochemical and histological analysis and 57 piglets from 21 farms were examined serologically. Possible causes for wasting other than PCV-2 were investigated by macroscopic, histological and bacteriological methods. PCV-2 antigen was identified immunohistochemically in the lymphatic organs in 11 of these 72 piglets. However, only 4 animals showed histological changes typical of PMWS. PCV-2 antibodies were found in 70% of the piglets. Piglets with wasting syndrome not associated with PCV-2 infection suffered from conditions including porcine proliferative enteropathy, gastric ulcers, polyserositis and polyarthritis. The most frequent condition was chronic enteritis not associated to circovirus infection. The results from the serological analyses indicate a wide distribution of PCV-2 in the Swiss pig population. However, confirmed cases of PMWS were rare in the investigated piglets. PMID- 15526603 TI - [Enzootic pneumonia (EP) in Swiss swine herds after area-wide eradication: epidemiological analysis of outbreaks occurring between 1999 and 2003]. AB - In a retrospective analysis, infections of swine herds with Enzootic Pneumonia (EP), occurring after the regional eradication programme had been completed, were evaluated and described according to epidemiological criteria. The aim of this study was to obtain interim results about status, progress and trends of the area wide eradication in Switzerland over all regions involved in the programme. The population comprised pig farms with eradication of EP completed by the end of 2002. Incidence of EP infection was calculated for the years 2000-2003. Seasonal effects, influences from production-type, herd size, and pig density in the surrounding area on incidences of EP outbreaks were investigated. In 2000-2002, annual incidences were steadily decreasing from 3.1% to 2.0%, and they showed great regional variation. In 2003, contrary to the long-term trend, a minor increase of overall incidence from 2.0% to 2.1% was observed while the incidence for breeding-only farms continued to recede. Possible explanations for this are discussed. Previous observations on EP infections accumulating in the cold season could not be confirmed. Large farms, finishing farms, and farms located in densely populated pig areas as well as farms being part of multisite production rings had a higher risk of infection. In spite of occurrence of EP outbreaks in eradicated areas, the current status of the area-wide eradication is promising, and a further decline in incidence of EP infection can be expected in the future. PMID- 15526604 TI - [Life threatening intestinal bleeding in a Bearded Collie associated with a food supplement for horses]. AB - In a Bearded Collie with acute weakness, hematemesis, melena, painful abdomen and pale mucous membranes a hematocrit of 13% and panhypoproteinemia were found. This combination of findings was the manifestation of severe gastrointestinal bleeding. Despite intensive laboratory and imaging investigations no systemic or local cause could be identified. After repeated client interrogation it was found that the dog had been receiving a food supplement for equines. It was further detected that this supplement besides a shell extract also contained willow (Salicaceae) and meadowsweet (Filipendula, Spiraea ulmaria) which contain salicin.Thus, the administration of this supplement was considered a possible cause of gastrointestinal bleeding. Even though measurement of toxic metabolites in the blood was not obtained and a cause-and-effect relationship not definitively proven, on principle it must be taken into consideration that any natural and so-called harmless agent supposed to have a positive effect may be associated with adverse effects in a predisposed individual. PMID- 15526605 TI - VP candidate John Edwards attends Labor Fest, meets with nursing leaders. PMID- 15526606 TI - RNs at University of Michigan Health System ratify new contract. PMID- 15526607 TI - My employer offered a settlement to my grievance ... should I take it? PMID- 15526608 TI - Statewide 'Hope for Haiti' project launched by nursing students. PMID- 15526609 TI - [Portuguese-Brazilian exchanges: challenges and opportunities]. PMID- 15526610 TI - [Epidemiology of heart failure in mainland Portugal: new data from the EPICA study]. AB - Heart failure (HF) is common, costly, disabling and lethal, and can be prevented. The progression of overt HF (CHF) can be slowed by correct management including precise diagnosis and appropriate therapy. Patients with CHF can be misdiagnosed, mainly in primary care, where patients are actually less symptomatic than those seen in hospitals. Accurate diagnosis requires objective evidence of cardiac dysfunction at rest by imaging techniques, according to the European Society of Cardiology (ESC) Guidelines. The EPICA Project (EPidemiologia da Insuficiencia Cardiaca e Aprendizagem) was one of the first European studies designed to evaluate the prevalence of CHF according to those criteria. The estimated overall prevalence of CHF in Portugal was 4.36% in adults over 25 years. 1058 individuals were identified by the Boston questionnaire as possible or probable CHF cases; only 551 had objective evidence of cardiac dysfunction at rest by echocardiography. 264 patients in sinus rhythm had a Boston score > 3 and no echocardiographic abnormalities. These patients were predominantly older obese women; coronary artery disease was less prevalent than in patients with proved CHF. The ECG was normal in 40.9%, versus 20% in CHF patients. About one half of these patients were prescribed ACE inhibitors and diuretics; 17% were prescribed digoxin. According to these EPICA data concerning CHF, older obese women are more frequently misdiagnosed and incorrectly medicated. PMID- 15526611 TI - [Heart failure in Brazil. The EPICA-Niteroi study]. AB - In this paper we review various problems of cardiovascular disease, particularly heart failure (HF), in Brazil. Cardiovascular disease is the main cause of death in Brazil, and heart failure is the most common cause of hospitalization in the Brazilian health system. The problem is even more serious GIVEN that the population is ageing rapidly; by 2025 the country will be ranked sixth worldwide in terms of numbers of old people. According to figures from DATA-SUS, HF is the most important cause of hospital admissions among the elderly in the Brazilian health system. The most frequent etiologies of HF encountered in clinical practice are coronary artery disease and hypertension. Another problem is the difficulty experienced by a large proportion of the population in gaining access to public health services; the health system is unable to cope with the needs of the population and the costs involved. An important reason for including HF under the public health system is the low socioeconomic level of many patients, which leads them to abandon treatment. PMID- 15526612 TI - [From diagnostic and therapeutic guidelines to clinical practice in Brazil]. AB - Heart failure (HF) is a clinical syndrome of high incidence and high prevalence all over the world. Available therapeutic options for chronic HF slow the progression of ventricular remodeling and decrease the number of arrhythmic deaths; the resulting increased life expectancy leads to a higher prevalence of heart failure in the community and a higher number of hospital admissions. In Brazil, the largest country in Latin America, hospital admissions due to HF represent approximately 4% of all hospitalizations and 31% of hospitalizations for cardiovascular diseases. Brazilian cardiologists are increasingly following, the Brazilian Society of Cardiology Guidelines on Heart Failure, which are very similar to other international guidelines. Differences between the public and private health systems in our country, although, compromise the compliance of patients to HF treatment, decreasing the efficacy of treatment in the lower social classes. PMID- 15526613 TI - [Heart failure management: guidelines versus clinical practice]. AB - In Europe the management of heart failure is largely conducted by primary care physicians and several reports have revealed differences between guidelines and actual practice. The IMPROVEMENT of Heart Failure project was designed to assess how patients with heart failure are managed by those physicians. Two surveys, the "perception survey" and the "actual practice survey" addressed to 1363 physicians, included questions on diagnosis, functional assessment and treatment. The major discrepancies found were: the lack of awareness about the concept of heart failure with preserved systolic function; the low number of echocardiograms requested; the low rate of prescription of beta-blocking agents and spironolactone; and the inappropriately low of angiotensine-converting enzyme inhibitors prescribed. PMID- 15526614 TI - [Heart failure clinics: the Brazilian experience]. AB - The treatment of congestive heart failure (CHF) is complex due to the need for multiple drugs and many surgical options. The accuracy, time of diagnosis, the introduction of treatment based on evidence, the detection of co-morbidities, and follow-up to improve prognosis are fundamental. Many studies have demonstrated that specialized centers in the treatment of CHF are superior and show better cost/benefit relation in comparison with usual treatment. The complexity of CHF treatment also requires a sub-specialist in CHF. The programs of CHF clinics include intensive education of family/patients and a careful follow-up performed by nurses and the multidisciplinary team. In a prospective randomized study in the CHF clinics of the Heart Institute (InCor) this program reduced the number of hospitalizations by 38%, the number of days during the hospitalizations by 71%, and necessity of emergency care by 27%. Published studies have demonstrated tendency or reduction of mortality, reduction of mortality associated with hospitalizations, reduction of hospitalizations, with sustained effects during a long-term follow-up and consequent reduction of costs. In the treatment of CHF it is necessary to include patients in programs of CHF clinics. PMID- 15526615 TI - [What causes edema, how can it be flushed out? Edemas require differentiated diuresis]. PMID- 15526617 TI - [After care for kidney transplant recipients: should the family physician really carry the major responsibility?]. PMID- 15526616 TI - [Edema of unknown origin. What helps the young woman with edematous legs?]. PMID- 15526618 TI - [Acute renal failure caused by analgesics. Successful treatment of knee pain, but patient died]. PMID- 15526619 TI - [Controversy over textiles impregnated with antimicrobial agents. Odor killers in socks and underwear: risk for the skin? (interview by Waldtraud Paukstadt)]. PMID- 15526620 TI - [Dyspepsia--tachycardia--dyspnea--How to recognize psychiatric disorders]. PMID- 15526621 TI - [Anxiety-ridden patients in the doctor's office]. AB - Initially, patients with anxiety syndromes consult the general practitioner complaining of physical symptoms. After an appropriate differential diagnostic work-up, the family doctor can therefore check whether the patient might possibly be suffering from an anxiety disorder. For the treatment of such disorders, psychotherapeutic and psychopharmacological strategies all have a role to play. The general practitioner can initiate supportive medication before referring the patient to a specialist. Early detection and treatment of an anxiety disorder can prevent the condition from becoming chronic and may also obviate the need for unnecessary repeated diagnostic measures and treatments. PMID- 15526622 TI - [Somatoform disorders--what must the general practitioner know?]. AB - Somatoform disorders are common conditions. Provided that the diagnostic criteria are known, however, they are not difficult to diagnose. In this connection, the family doctor has it within his power, by initiating empathic client-centred (conversation) therapy with the patient, to create a basis for a confidence building and a long-lasting physician-patient relationship. In addition to a psychotherapeutic intervention or psychodynamic treatment or behavioral treatment, we now have the first positive results of a psychopharmacological treatment regimen with opipramol and the St. John's Wort extract LI 160. PMID- 15526623 TI - [Adult attention deficit/hyperactivity disorder]. AB - ADHD (attention deficit/hyperactivity disorder) in adults is a more complex pathological condition than ADHD seen in children and adolescents. The number of reports of impaired self-regulation are on the increase. Psychiatric comorbities are being found ever more frequently, and negative life experiences are coloring the clinical presentation to an ever greater extent. Therapeutic strategies involving the use of stimulants and antidepressants are often needed to pave the way for individual and group psychotherapy. Despite the fact that it is currently considered to be "fashionable", the diagnosis of ADHD is a clinically relevant and persisting psychological disorder. PMID- 15526624 TI - [Hematemesis]. PMID- 15526625 TI - [Obesity and the metabolic syndrome in children and adolescents]. AB - Childhood obesity is a major risk factor, especially for the metabolic syndrome and further cardiovascular diseases. The metabolic syndrome is defined by a combination of obesity, arterial hypertension, hyperlipidemia and dyslipidemia, insulin resistance or type 2 diabetes mellitus. Obesity in early childhood further increases the risk of premature illnesses and early death, therefore raising public health concerns. About one third of overweight or obese children and adolescents exhibit features of early metabolic syndrome. In view of the increasing issue of early-onset obesity, effective family- and school-based primary care programs in addition to sufficient weight-reducing therapeutic approaches are necessary in order to achieve a modification of our sedentary lifestyle. Local long-term therapeutic programs for obese young patients are an important tool to establish a more active and health-oriented life style in children and their families and to reduce the individual and public burden of obesity in childhood. PMID- 15526626 TI - [The efficacy of a combination herbal medicine in the treatment of functional dyspepsia]. PMID- 15526627 TI - [KBV faulted: there is not enough money for drugs. They have caused the deficit for physicians!]. PMID- 15526628 TI - [Supra-regional cooperation will soon be possible. Do you know the influential colleagues?]. PMID- 15526629 TI - [Establishing a medical care center: is it financially beneficial?]. PMID- 15526630 TI - [Disposition court decides: a physician may also advertise with drug names]. PMID- 15526631 TI - [Thermotherapy. Rusty iron for back pain? (interview by Dr. Marianne Kupfer)]. PMID- 15526632 TI - [New STIKO recommendation. Varicella vaccination for all]. PMID- 15526633 TI - [Tightening the belt! "Spare tires" promote insulin resistance]. PMID- 15526634 TI - [Motivational therapy can fail here. Sleep disorders in depressions]. PMID- 15526635 TI - [Despite drug treatment, ever second hypertensive patient with morning blood pressure too high]. PMID- 15526636 TI - [PTCA with drug eluting stent. 90% fewer restenoses in high risk patients]. PMID- 15526637 TI - [Phytotherapy tested in heart-healthy seniors. More vital with Crataegus monogyna]. PMID- 15526638 TI - [Low HbA1c value is not a reason for optimism. Infarct risk due to postprandial glucose peaks]. PMID- 15526639 TI - [Werner Forssmann tested the first heart catheter on himself. For this reason he was fired by the chief physician]. PMID- 15526640 TI - [Severe toe pain after heart catheterization]. PMID- 15526642 TI - [Pulpotomy of deciduous teeth]. AB - Rather easy to perform, pulpotomy of the deciduous teeth is the most frequent endodontic treatment performed on children, but also the most controversial. Based on the amputation of the pulp chamber and the conservation of the inflammation-free root canals, the clinical results can be good, depending on the materials used. In this, calcium hydroxide (Ca(OH)2) as well as zinc-oxide eugenol (ZOE) have been proven very inflammatory whereas Formocresol (FC) remains the reference even if its clinical toxicity is still reported in literature on a very controversial way. Nevertheless, this was sufficient to trigger and stimulate a search for alternatives, and led to the proposition to use ferric sulfate and even more recently MTA as new bases for the treatment of the pulp stumps after pulp chamber amputation. PMID- 15526641 TI - [Clinical evaluation of the INJEX system, a local anesthesia system without needles: a comfort evaluation study]. AB - The use of needless local anaesthesia systems in dentistry can be of a help in treating needle phobia patients. The aim of this comfort study was to compare a classical local infiltration anaesthesia with a needle-free system. INJEX (ROSCH AG Germany). A split mouth design study was performed. All patients in this study needed 2 restorations. In order to receive objective information on both systems needle-phobia patients were banned. Therefore the first restoration was performed with the classical system (with needle) and the well-acceptance was evaluated. The second restoration was done with the needle-free system. Both patient and dental practitioner performed an evaluation after each treatment. Evaluation by the patient was given by a questionnaire on the comfort of the treatment using Faces Pain Scale, Lickert Scale and a modified version of the Abbreviated Acceptability Rating Profile. By the dental practitioner the comfort of the treatment and the amount of local anaesthesia needed was evaluated. The study was approved by the ethical committee of Ghent University Hospital Belgium. CONCLUSION: The INJEX system can be a valuable alternative to use in paediatric dentistry, although non needle phobia patients in this study did not preferred the needle free INJEX system above the classical local injection. PMID- 15526643 TI - [Calcium hydroxide and treatment of inflammatory inter-radicular bone resorption of non-vital deciduous molars]. AB - On non-vital deciduous molars, inter-radicular bone resorption is often an indication of extraction. The endodontic treatment of these teeth by means of zinc oxide-eugenol (ZOE) paste also showed its limits. To mitigate the deficiencies of this material, we suggested a preliminary treatment by means of calcium hydroxide (Ca(OH)2) for its anti-inflammatory and antiseptic properties as well as its ability to stimulate calcified tissues apposition or remineralisation. This study concerns 21 non-vital deciduous molars. X-rays excluded any lesion of the underlying permanent bony crypt (bone tissue) as well as any inflammation of the dental follicle. After preparation, root canals were filled by means of Pulpdent. An initial X-ray check was made 15 days and then every 3 months. After disappearance of the inflammatory resorption, root canal fillings were performed with ZOE paste. The remineralisation of the inter radicular alveolar bone was observed for 14 deciduous molars, which were then filled using ZOE. The remineralisation period varies from 3 to 18 months depending on the scale of the lesion. Of the 7 failed treatments, 3 failed following downfall of the crown filling material, and 2 due to failure to keep appointments and late replacement of resorbed Ca(OH)2. On 2 teeth, the treatment did not stop the lesion forming. Calcium hydroxide (Ca(OH)2) give encouraging results in the treatment of inter-radicular alveolar bone resorption of non-vital deciduous tooth. Its fast resorption requires rigorous controls, frequent refills, and thus strong motivation on the part of the child and parents. It cannot, on any account, be considered as permanent filling material. PMID- 15526644 TI - [Early recognition of orthodontic problems by the general dentist during oral examination: signs which should attract attention]. AB - The last decennia more and more patients are aware of the advantages of a good dental health. Dental minded parents in their children visit the dentist (general practitioner, GP) on a regular basis for a routine check up. During this check up, signs of possible future orthodontic problems can be encountered by the GP. In this article guidelines are handed to the GP by means of a flow chart which can be used at the end of the dental examination. Anterior/posterior relation, cross-bites, deep and open bite, habits and crowding shall be briefly discussed. Heaving knowledge of these, the GP can decide referring the patient to the orthodontist for further orthodontic evaluation. PMID- 15526645 TI - [Prosthetic rehabilitation and accompaniment of growth in cases of early tooth loss in children]. AB - Teeth, in the general phenomenon of growth contribute to the blooming and the growth of the young child. It plays a paramount role in the communication while taking part in the development of the language and of the phonation, and by perfecting the shape of the face. The prosthetic rehabilitations are addressed to children who present more or less significant early losses of teeth, whose origin could be congenital, structural, post-decays or traumatic, and bearing on temporary, mixed or final teeth. The series of case presented clarifies the methods which we employ in our service for the design and the practical realization of the prosthetic rehabilitations. The dental lose seriously disturbs the physical development of the child, it can slow down its school development and affect in a pathological way its relational life. The installation of prosthesis makes it possible to the child to feel like the others, to eat normally, to better sleep, to be able to present its face without causing mocking remarks. Among our patients candidates with the prosthetic rehabilitation, the children in low-age are most numerous and present the highest frequency of the traumatisms. Their first real smile is already the justification of the decision of the specialist. Nevertheless, the number of toothless children who profit from this type of rehabilitation is very small. PMID- 15526646 TI - [Dental symptomatology associated with connective tissue anomalies]. AB - Subjects affected with inherited disorders, of the connective tissue make up an important population, carrying high risks as to distinct aspects of oral health and dental treatment. These generalized conditions may produce serious clinical symptoms in different orofacial structures, which have to be dealt with, or anticipated, when considering dental treatment. The most prevalent disorders result from deficiency of Type I collagen, an important extracellular matrix protein regulating both the structural and mechanical properties of most of the orofacial tissues. Recurrent jaw fractures, an increased liability for development of temporomandibular disorders, periodontal disease and mucosal fragility, an abnormal tooth color and/or shape, and pulp obliteration may feature as major clinical manifestations of the respective disorders. Deficiency of fibrillin, a protein providing soft tissues with elastic capacities, may produce a long face with a high and narrow palate, an increased liability for the developnet of temporomandibular disorders and periodontal disease, and root dsyplasia. Whenever present, these manifestations/risk factors have to be integrated in dental treatment strategies. In cases with high risk for cardiovascular complications, specific preventive measures, such as cardiac output monitoring and the administration of appropriate local anesthetics, have to be taken before starting any invase dental treatment. The present paper aims to provide the practitioner with an appreciation of the most prevalent inherited disorders of the connective tissue with their respective genetics, molecular aspects of pathology, medical and oral manifestations, and guidelines for dental treatment. PMID- 15526648 TI - [What is the relation between the presence of caries in the deciduous dentition and the chronology of the eruption of the permanent teeth?]. AB - Based on data obtained from a prospective epidemiological study in a random sample of 4468 Flemish children, accurate emergence ages for permanent teeth could be established. When data on permanent tooth emergence in different populations are compared, differences are most pronounced for premolars. Several authors hypothesised that this difference could be explained by a difference in caries experience in the primary molars. The purpose of this study was to quantify the effect of caries experience in a primary molar on the timing of emergence of its successor. The results indicate that the emergence of the premolars was accelerated by 2 to 8 months when its predecessor had been decayed and or filled but had not been extracted. Premature loss of maxillary primary molars resulted in a significant acceleration of the emergence of the premolars; this was not observed in the mandible. In conclusion, when considering permanent tooth emergence ages, caries experience in the primary dentition should be taken into account. PMID- 15526647 TI - [Use of osteoconductive materials in pediatric dental medicine]. AB - Osteoconductive materials are well known to have the ability to form a characteristic scaffold for bone replacement. The use of these agents is mostly described in periodontal procedures. The present study deals with the use of two different osteoconductive materials used for repairing bone defects following traumatic injuries. The materials used were Bio-Oss (Geistlich) an allogenic demineralised bone and Fisiograft (Ghimas) a synthetic co-polymer of polyglicolic and polylactic acid. These materials were used in five different clinical cases with the objective to compare their typical properties during application and to evaluate radiographically and clinically their healing ability. In three cases filling of the alveolus was performed after extraction of traumatised incisors as a consequence of root resorption or fracture; two patients suffered from periodontal defects. All cases were followed every three months for at least two years and maximum 6 years. Regarding the handling of the materials all products were well accepted. From our clinical experience all modalities are found to be suitable. After clinical and radiographical evaluation normal bone formation was found in all cases without any complaints nor clinical symptoms. These clinical findings support the use of osteoconductive agents in the field of Paediatric Dentistry in well-defined indications. PMID- 15526649 TI - [Not only diabetics have experienced the pain... distressing neuralgia]. PMID- 15526650 TI - [Evidence-based management of diabetic polyneuropathy. "Burning feet" alarm signal]. PMID- 15526651 TI - [Sudeck hyperalgesia and phantom pain. Extreme pain can be conquered with exercise]. PMID- 15526652 TI - [Immunosuppressed patient becomes pregnant. Termination of pregnancy unavoidable?]. PMID- 15526653 TI - [Review of European national soccer competition. Could restless legs have saved Rudi?]. PMID- 15526654 TI - [Digital mammography. An alternative to current screening?]. PMID- 15526655 TI - [Alternative medicine practitioner promised healing. That is why my patient had to die at 29 years of age]. PMID- 15526656 TI - [When the mountain calls: who is at risk at high altitude? (interview by Dr. Judith Neumaier)]. PMID- 15526657 TI - [Therapeutic training: power, balance and weight control not merely for athletes. More exercise for everyone]. PMID- 15526659 TI - [Current views on the prevention of ailments of the musculoskeletal system through muscle strengthening programs]. AB - Because of an increasingly older population structure and the enhanced relevance of body composition, the prevention of problems arising from the musculoskeletal system has become of central importance. Strengthening and muscle-building training have been proven to be useful even for older people. The training is focused on the types of movement and exertion that frequently occur in the "activities of daily living." In addition, a purely hypertrophic training for building muscle mass must be differentiated from a more sensorimotor oriented training with situational employment of the muscles for optimizing motor control and coordination. There are no contraindications for strength training when the program is adapted to the individual. The rate of injury is small, even for people at low performance levels. PMID- 15526658 TI - [Current views on the importance of physical activity]. AB - To improve the individual health profile and to solve the growing overweight problem, a long-term change in the lifestyle to one which includes an energetically balanced combination of diet and activity is essential. Physical activity and the muscles involved are the primary means by which body composition and energy turnover are regulated. A state of imbalance has decisive consequences on the development of atherogenic and inflammatory risk factors. Additionally, the aging process is significantly influenced by the long-term retention or loss of muscle mass. The Deutsche Gesellschaft fur Sportmedizin und Pravention (German Society for Sports Medicine and Prevention, a registered association) offers within the concept of a therapeutic lifestyle change, an educational program for overweight adults (M.O.B.I.L.I.S.). PMID- 15526660 TI - [Physical activity and dyslipoproteinemia]. AB - The positive influence of regular physical activity on lipoprotein metabolism and, hence, on the risks for atherogenesis has been documented in controlled studies. Although the levels of total and LDL cholesterol (LDL) usually change only slightly through physical activity, there is a clear change in the LDL composition with a reduction in the atherogenic small dense LDL particle. An activity-induced increase for HDL cholesterol (HDL) between 4 and 29% and a reduction in the triglyceride (TG) level between 4 and 37% are described. To achieve this, it is necessary to increase energy consumption by 1000-1200 kcal/week and to attain an energy consumption of ca. 2500 (> 2000) kcal/week, optimum ca. 3500 kcal/week. PMID- 15526661 TI - [Otitis caused by Vibrio cholerae non-01/non-0139 strains acquired in Germany]. PMID- 15526662 TI - [Preventing end-organ damage in hypertension. What is the significance of the VALUE Study for general practice?]. PMID- 15526663 TI - [Indications for antihypertensive therapy. Trans-Atlantic differences]. PMID- 15526664 TI - [Outpatient treatment of patients suffering from Alzheimer dementia. Two exploratory studies on the characteristics of therapy in medical practice]. PMID- 15526665 TI - [How nose-blowing assistants promote their general practice]. PMID- 15526666 TI - [Not every colleague is happy about it. In emergencies now only 1 co-pay]. PMID- 15526667 TI - [Drug costs should be lower. Hessian colleagues beaten without compassion]. PMID- 15526668 TI - [Are you planning a cultural vacation? Caution in old masonry!]. PMID- 15526669 TI - [Medicine needs new concepts. Prevention for the future (interview by Dr. Dirk Einecke)]. PMID- 15526670 TI - [Overactive bladder. New anticholinergic drug controls urinary urge]. PMID- 15526671 TI - [Optimizing antihypertensive therapy. In 5 years 35,000 strokes are preventable]. PMID- 15526672 TI - [Nasal obstruction on the decline. Antihistamine allows allergic patients to breathe]. PMID- 15526673 TI - [Decreasing LDL level moderately or drastically. Is differential therapy with statins available? (interview by Dirk Einecke)]. PMID- 15526674 TI - [ASS plus dipyridamole instead of ASS monotherapy. Risk for second stroke cut in half]. PMID- 15526675 TI - [Depressed, lacking motivation, unconcentrated. Signs of central deficiency of noradrenaline]. PMID- 15526676 TI - [Type 2 diabetic patient with kidney damage. Sartans banish dialysis risk]. PMID- 15526677 TI - [Consulting the ophthalmologist for assessment]. PMID- 15526678 TI - [10 minute consultation: urticaria]. PMID- 15526679 TI - [Dermatologist becomes hepatologist]. PMID- 15526680 TI - [Medicine of the 21st century and cell biology]. PMID- 15526681 TI - [Tissue cell systems as a basis for the biomedical cell technologies of the next generation: ideology outlook]. PMID- 15526682 TI - [Ethic issues of stem cells]. PMID- 15526684 TI - [Stem cells: present-day, problems, outlooks]. PMID- 15526683 TI - [An efficiency study of transplantation of embryonic nervous-tissue preparations in experimental 6-OHDA-induced hemiparkinsonism]. PMID- 15526685 TI - [The concept of cell therapy in autoimmune diseases]. PMID- 15526686 TI - [Experimental substantiation of the efficiency of using the stem and progenitory cells of the bone marrow for regulating the recovery processes in organs in acute and chronic affections]. PMID- 15526687 TI - [Cell and interactive technologies in the treatment of congenital and acquired heart diseases and ischemic heart disease]. PMID- 15526688 TI - [Cell technologies in the therapy of malignant neoplasms]. PMID- 15526689 TI - [Cell technologies in oncology]. PMID- 15526690 TI - [Transplant versus leukemia reaction]. PMID- 15526691 TI - [Production of cell cultures from mesenchymal stem cells of the human bone marrow and their use in medicine]. PMID- 15526692 TI - [The reality and problems related with the use of stem cells in clinical practice]. PMID- 15526694 TI - [Resolution of the XIII (76) session of the General Assembly of the Russian academy of medical sciences "Cell technologies in medicine"]. PMID- 15526693 TI - [Resolution of the XIII (76) session of the General Assembly of the Russian academy of medical sciences on the report made by the Presidium of the Russian academy of medical sciences (March 2001-September 2003)]. PMID- 15526695 TI - [Alexandr Vasilyevich Vishnevsky (on the occasion of his 130th birthday anniversary)]. PMID- 15526696 TI - Keeping surgical procedure turnover in perspective. PMID- 15526698 TI - The new barrier standard--whose interests does it serve? PMID- 15526699 TI - From handmaiden to right hand--World War I--the mud and the blood. AB - By the beginning of World War I, modern innovations and advances in the field of medicine were common. Physicians knew about bacteria, how disease spread, and the importance of antisepsis. To prepare for the war, Great Britain developed elite corps of army nurses, fully-equipped military hospitals, and trained personnel who established field hospitals and base hospitals. Alice Fitzgerald, a nurse from Baltimore, was drawn into the conflict when she was asked to serve as the Edith Cavell Memorial Nurse with the British Army. PMID- 15526700 TI - Intraoperative use of autologous platelet-rich and platelet-poor plasma for orthopedic surgery patients. AB - As use of autologous platelet-rich plasma (PRP) and platelet-poor plasma (PPP) increases for intraoperative care of a variety of patients, it is important for perioperative nurses to recognize their benefits. Autologous PRP may decrease postoperative drainage, reduce narcotic requirements, and facilitate an early return to mobility. Postoperatively, patients should experience fewer complications, recover more rapidly, and have a reduced hospital stay. This article defines autologous PRP and PPP, describes processing and application of PRP and PPP, and reports clinical outcomes of the use of platelet concentrate for a group of patients who underwent total knee arthroplasty. PMID- 15526701 TI - Treating patients with smallpox in the operating room. AB - Recent events around the world have emphasized the need for health care facilities to prepare to deal with biological threats, including smallpox. At Walter Reed Army Medical Center, Washington, DC, administrators recognized the need for a policy on handling patients with smallpox in the OR and asked a group of students to create a template policy for care of patients with smallpox in need of surgery. This article provides a brief history of smallpox, concerns surrounding smallpox today, and smallpox characteristics with which perioperative personnel should be familiar, as well as a guideline for treating patients in the OR who have smallpox. PMID- 15526702 TI - Preparing RNs for the OR through a certificate in perioperative nursing program. AB - The need for qualified perioperative nurses is an ongoing issue, and many hospitals are training their own. Often, a facility will invest time and money to train a perioperative nurse, only to have the nurse leave to work for a competitor. This article describes how hospitals in the city of Louisville, Ky, pooled resources to develop a certificate in perioperative nursing program to address these issues. PMID- 15526703 TI - A knowledge comparison of nursing students in perioperative versus other rotations. AB - As more patients undergo surgical interventions in ambulatory facilities and patients in acute care settings have shorter lengths of stay, nursing students may have limited interaction with the surgical patient population. This study used a repeated measure, experimental design to compare knowledge of surgical patient care among undergraduate nursing students who experienced a perioperative clinical rotation (ie, experimental groups) with knowledge among students who experienced rotations in other clinical settings (ie, control groups). Results demonstrated a significant change in mean knowledge scores over time and a significant difference in mean scores between the experimental and control groups. Students in the perioperative group scored higher in all four testing periods. PMID- 15526704 TI - Creating a successful RN first assistant practice. AB - Many RN first assistants (RNFAs) aspire to having their own practice. This article provides helpful exercises to help RNFAs determine if they have what it takes to be business owners and their own bosses. The pros and cons of different business structures-sole proprietorships, partnerships, and corporations-also are discussed to help RNFAs determine the best structure for their independent practice. PMID- 15526705 TI - Perioperative surgery in the twenty-first century--two case studies. AB - Comprehensive perioperative information technology (IT) systems can help health care organizations and their ORs reduce costs, increase revenues, improve processes, and enhance caregiver and patient satisfaction. This article examines what perioperative IT systems have done for two health care organizations and details implementation, the benefits these hospitals derived, next steps, and lessons learned. PMID- 15526706 TI - AORN guidance statement: safe specimen identification, collection, and handling in perioperative practice settings. PMID- 15526707 TI - Human patient simulation: a teaching strategy. PMID- 15526708 TI - The anti-platelet approach targeting the fibrinogen ligand of the GPIIB/IIIa receptor. AB - Activation of the platelet surface receptor GPIIb/IIIa is the final pathway of platelet aggregation, regardless of the initiating stimulus. RGD analogues, peptidomimetics and monoclonal antibodies to GPIIb/IIIa have been developed targeting the blockage of the receptor and inhibition of the fibrinogen binding. However, the intrinsic activating effect of GPIIb/IIIa blockers is widely discussed as one potential contributing factor for the disappointing outcome of trials with GPIIb/IIIa inhibitors. An alternative method for thrombus prevention could be the use of specific fibrinogen blockers since they will act at the final step of the platelet aggregation and are expected to leave the receptor unaffected. To achieve this target the design of the fibrinogen ligands could be based on (i) sequences derived from GPIIb/IIIa ligand binding sites, and (ii) sequences complementary to RGD and/or to fibrinogen gamma-chain. The available information, which could be used as a starting point for developing potent fibrinogen ligands, is reviewed. PMID- 15526709 TI - Polymer-bound alkyltriazenes for mild racemization-free esterification of amino acid and peptide derivatives. AB - A novel tool for polymer-assisted solution phase (PASP) esterification of amino acid and peptide derivatives has been developed. When treated with carboxylic acids, polymer-bound alkyltriazenes react with a loss of nitrogen and transfer of the alkyl moiety to the carboxylate anion to form the corresponding alkyl esters. There are no limitations with regard to either the protecting groups or the nature of the amino acid. Furthermore no racemization occurs at the chiral centers of the amino acids as demonstrated by chiral GC-MS analyses. Alkyltriazene-resins were also applied successfully to the esterification of peptide acids and other peptidic structures, such as tripalmitoyl-S-glyceryl cysteine (Pam3Cys). The triazene-mediated esterification reaction is exceptionally mild, and there is no need for prior activation of the carboxy groups. This method is therefore particularly suitable for the alkylation of complex peptidomimetic structures prone to racemization and for acid-sensitive structures. PMID- 15526710 TI - Interaction between amyloid beta-protein aggregates and membranes. AB - The conversion of soluble, nontoxic amyloid beta-protein (Abeta) to aggregated, toxic Abeta rich in beta-sheet structures is considered to be the key step in the development of Alzheimer's disease. Therefore, extensive studies have been carried out on the mechanisms involved in Abeta aggregation and the characterization of Abeta aggregates formed in aqueous solutions mimicking biological fluids. On the other hand, several investigators pointed out that membranes play an important role in Abeta aggregation. However, it remains unclear whether Abeta aggregates formed in solution and membranes are identical and whether the former can bind to membranes. In this study, using a dye-labeled Abeta-(1-40) as well as native Abeta-(1-40), the properties of Abeta aggregates formed in buffer and raft-like membranes composed of monosialoganglioside GM1/cholesterol/sphingomyelin were compared. Fourier transform infrared spectroscopic measurements suggested that Abeta aggregates formed in buffer and in membranes have different beta-sheet structures. Fluorescence experiments revealed that Abeta aggregated in buffer did not show any affinity for membranes. PMID- 15526711 TI - NMR studies of peptide T, an inhibitor of HIV infectivity, in an aqueous environment. AB - The synthetic octapeptide peptide T (ASTTTNYT) has been shown to interfere with binding of the HIV-1 envelope glycoprotein gp120 to the chemokine receptor R5, thus preventing viral infection. This study investigated the degree of conformational order of two analogs of peptide T, one biologically active (D-Ala peptide T amide) and one inactive (D-Ala, D-Tyr peptide T amide) using nuclear magnetic resonance (NMR) spectroscopy in an aqueous environment, both in solution and in the frozen solid state. Standard solution NMR techniques such as DQFCOSY, HMQC, ROESY and inversion recovery measurements have been utilized to characterize these peptides. Solid state NMR experiments were likewise employed to study the peptides in a frozen glycerol:water mixture. The NMR results indicate that the monomeric form of both peptide T analogs have considerable conformational heterogeneity. Solid state NMR studies indicate aggregation of D Ala peptide T, possibly into a beta-sheet structure, at concentrations higher than 10 mM. PMID- 15526712 TI - High-yield, solid-phase synthesis of humanin, an Alzheimer's disease associated, novel 24-mer peptide which contains a difficult sequence. AB - Humanin is a novel, 24-mer residue bioactive peptide, which antagonizes Alzheimer's disease (AD) related neurotoxicity and offers a hope for developing new therapeutics against AD. Access to adequate amounts of pure humanin is a prerequisite for further, thorough, investigation of the pharmacological properties and therapeutic potency of the peptide. Until now, humanin has been obtained mainly by molecular biology techniques. In this work the Fmoc solid phase synthesis of humanin on an in-house prepared 2-Cl-tritylamidomethyl polystyrene resin is described fully. Special precautions, i.e. prolonged deprotection steps, should be taken to achieve a high overall yield, since humanin seems to contain a 'difficult sequence' (R4G5F6S7C8L9) near its highly lipophilic, biologically important region L9L10L11L12. PMID- 15526713 TI - Effect of humanin analogues on experimentally induced impairment of spatial memory in rats. AB - Humanin and its analogues have been shown to protect cells against death induced by various Alzheimer's disease genes and amyloid-beta-peptides in vitro: the analogue [Gly14]-humanin has also been shown to be potent in reversing learning and memory impairment induced by scopolamine in mice in vivo. It is important to validate these results by using other behavioral methods. In this study, the effect of [Gly14]-humanin and des-Leu-PAGA, another analogue (0.2 micromol kg( 1), i.p.) on the 3-quinuclidinyl benzilate-induced (2 mg kg(-1), i.p.) impairment of spatial memory in the multiple T-maze in rats has been evaluated. Both peptides reversed the impairment of spatial memory. These results indicate the potential of humanin analogues in modulation of the cholinergic system. PMID- 15526714 TI - Gender role across development in adult women with congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - This study evaluated the degree of femininity and masculinity at different developmental stages in a group of adult women, some of whom were exposed to elevated prenatal adrenal androgens as a result of congenital adrenal hyperplasia (CAH) due to 21 hydroxylase (21-OH) deficiency. Women who had presented to the Johns Hopkins Hospital Pediatric Endocrine Clinic for treatment of CAH due to 21 OH deficiency were included. The control group consisted of sisters of CAH participants and women referred for evaluation of polycystic ovary syndrome. Study participants were given a questionnaire asking them to indicate their degree of masculinity and femininity during childhood, adolescence, and adulthood. In addition, participants were asked questions related to their play behavior during childhood, including playmate preferences, toy preferences, and admiration of male or female characters during fantasy play. Across participant groups, self-reported femininity decreased in a dose response manner, according to prenatal androgen exposure. For all groups, femininity increased through developmental stages. Women with salt-losing CAH remained less feminine than controls into adulthood. Conversely, self-reported masculinity increased in a dose-response manner, according to prenatal androgen exposure, across participant groups. Women with CAH showed a decrease in masculinity across developmental stages, such that by adulthood, there were no significant differences in masculinity between controls and the women with CAH. Women with salt-losing CAH were more likely to recall preferences for boy playmates, male-typical toys, and admiration for male characters during childhood than other study participants. Our data support the effect of both prenatal androgen exposure and socialization on gender role behavior in adult women with CAH due to 21-OH deficiency. PMID- 15526715 TI - Hyperinsulinaemic hypoglycaemia in infancy and childhood--resolving the enigma. AB - Children with severe hypoglycaemia due to persistent hyperinsulinism in infancy (HI) generate some of the most formidable problems of management in contemporary paediatric endocrinology. Until recently its pathophysiology was an enigma, although it was thought to be due to an anatomical abnormality in the islets of Langerhans (so called 'nesidioblastosis'). During the last 6 years there has been an explosion of knowledge providing fundamental insights into the pathological mechanisms underpinning the abnormal insulin secretion. This knowledge has been facilitated by ENRHI, a programme of research funded by the European Union, which brings together clinicians and basic scientists from 14 different countries. This collaboration encompasses clinical paediatric endocrinology, intracellular biochemistry, membrane physiology and molecular biology. This collaboration has resulted in numerous publications generating new insights into the pathophysiology of HI and represents a paradigm for collaboration in paediatric endocrinology. This review article is based on a plenary lecture delivered at the European Society for Paediatric Endocrinology meeting in Montreal on behalf of the European Network for Research into Hyperinsulinism of Infancy (ENRHI). PMID- 15526716 TI - Adopted girls with idiopathic central precocious puberty: observations about character. AB - The aim of this study was to investigate the psychological traits of adopted girls affected by idiopathic central precocious puberty (ICPP). Twelve adopted girls were evaluated before, during GnRH agonist (GnRHa) treatment, and after the treatment was stopped. Before, during and after treatment, adopted girls with ICPP showed a normal degree of anxiety, an adequate level of maturity, and a correct sexual identification. None of them had symbiotic character traits as previously reported in non-adopted girls with ICPP. The perception of their body was poor before treatment, but improved during GnRHa treatment and psychological support. The changes in the life of adopted girls are so numerous that precocious puberty may represent an additional risk for psychological discomfort. Therefore to treat precocious puberty it may not only be necessary to prevent premature changes in their body but also to give them enough time to face up to their emotional and psychological problems and to become competent adolescents. PMID- 15526717 TI - Serum leptin levels in premature pubarche and prepubertal girls with and without obesity. AB - Leptin can be regarded as a marker of the nutritional status of the body. This study was performed to determine the correlation of leptin levels with insulin (I) and androgens in girls with premature pubarche (PP) and prepubertal controls (C) with (OB) or without (nOB) obesity. We studied 25 girls with PP and 14 C; girls were dived into two subgroups according to body mass index (BMI): OB (18 PP and 8 C) and nOB (7 PP and 6 C). Obesity was defined as BMI >95th percentile for chronological age. Serum levels of leptin, I, glucose (G), DHEAS, testosterone, androstenedione (A), cortisol, SHBG, IGFBP-1 and lipid profile were measured. The fasting G to I ratio (FGIR) was calculated and FGIR <7 was considered as suggestive of I resistance (IR). Data were analyzed comparing PP vs C and OB vs nOB. Serum DHEAS (0.60 +/- 0.45 vs 0.18 +/- 0.22 microg/ml) and A (895.5 +/- 420.4 vs 457.0 +/- 352.1 pg/ml) levels were significantly higher in PP than C. Other hormonal and metabolic parameters were similar. Serum leptin (30.8 +/- 18.3 vs 8.1 +/- 5.9 ng/ml), A (841.8 +/- 471.1 vs 522.5 +/- 317.2 pg/ml), DHEAS (0.53 +/- 0.44 vs 0.31 +/- 0.39 microg/ml), G (88.4 +/- 8.8 vs 80.2 +/- 8.1 mg/dl), I (13.5 +/- 7.7 vs 5.1 +/- 3.7 microU/ml) and total cholesterol (TC) (180.5 +/- 30.9 vs 161.8 +/- 29.5 mg/dl) levels were greater in the OB than in the nOB group. IR was observed in 10 girls with OB and in one with nOB. Leptin was correlated with BMI (r = 0.83), SHBG (r = -0.44), IGFBP-1 (r = -0.47), I (r = 0.37), A (r = 0.48) and TC (r = 0.36), but in multiple regression analysis only with BMI (r2 = 0.72, p < 0.001). Girls with PP and prepubertal OB girls showed elevated leptin levels independent of I and androgen levels. Girls with OB had a greater degree of hyperandrogenism and IR. As obesity, IR and hyperandrogenism are common findings in polycystic ovary syndrome (PCOS), which is more prevalent in young women with a history of PP, a role of leptin in PCOS can be suggested. In addition, girls with PP could be considered a population at risk for plurimetabolic syndrome. PMID- 15526718 TI - Hyperprolactinemia in children during the peripubertal period--personal observations. AB - There can be many reasons for functional hyperprolactinemia, including polycystic ovary syndrome (PCOS), obesity, insulin resistance and hypoglycemia. Prolactinoma is also a frequent cause. This study included 12 patients (10 girls and 2 boys) aged 14-17 years with hyperprolactinemia. Six patients were treated for PCOS, two for type 1 diabetes mellitus, and one patient suffered from Prader-Willi syndrome. In all patients, TSH, fT3, fT4, FSH, LH, testosterone, and E2 levels were measured. MRI of the pituitary was performed. Prolactin was measured with the metoclopramid dynamic test (MTC). IRI, C-peptide, oral glucose test, and minor pelvis ultrasound examination were performed in patients with PCOS. Pituitary adenoma was diagnosed in four out of 12 patients. In the remaining eight patients, functional hyperprolactinemia was found. All of these patients remain under pharmacological treatment with positive clinical results. CONCLUSIONS: 1. It is necessary to determine prolactin in each girl with unexplained amenorrhea irrespective of galactorrhea. 2. In some patients with prolactinoma the basal prolactin levels may be in the normal range, but they are increased in the MTC test. 3. In girls with various disorders of the menstrual cycle it is necessary to determine the level of prolactin with a provocative test. PMID- 15526719 TI - Cardiac mass and function, carotid artery intima media thickness, homocysteine and lipoprotein levels in children and adolescents with growth hormone deficiency. AB - INTRODUCTION: Early atheromatic changes have been found in the carotid and the femoral arteries of young adults with growth hormone (GH) deficiency (GHD). It has been shown that adults with GHD have increased cardiovascular risk factors. The aim of this study was to compare atherosclerotic risk factors in children with GHD and a normal population. PATIENTS AND METHODS: Twenty-five patients (10 girls, 15 boys) with GHD qualified for GH replacement treatment were included in the study. The control group consisted of 22 healthy children (11 girls, 11 boys) with no family history of atherosclerosis and cardiovascular disease. Cardiac mass and function was evaluated by one-dimensional, two-dimensional echocardiography and the Doppler method. Common carotid intima-media thickness was measured with a linear probe (7.5 MHz). Lipid metabolism parameters (total cholesterol, HDL- and LDL-cholesterol, triglycerides), lipoprotein A (Lp(a)) and homocysteine levels were measured. RESULTS: Cardiac dimensions and systolic parameters were within normal values in both groups. The mean index of left ventricular mass (68.37 +/- 18.64 g/m2) in patients with GHD did not differ significantly compared to controls (68.48 +/- 15.56 g/m2). No significant differences between the study and control group were observed when comparing systolic and ejection fraction values. Significantly higher mean values of IVRT parameter were found (70.8 +/- 14.2 ms vs 64.09 +/- 8.54% ms; p < 0.05). Other parameters characterizing systolic function were not significantly different. The mean concentration of total (178.28 +/- 31.1 mg/dl) and LDL-cholesterol (106 +/- 28.68 mg/dl) was significantly higher in patients with GHD than in controls (157.59 +/- 22.39 mg/dl, 84.54 +/- 22.01 mg/dl; p <0.05). Lp(a) (40.34 +/- 12.45 mg/dl vs 11.02 +/- 4.82 mg/dl; p <0.05) and apolipoprotein B (ApoB) (71.12 +/- 18 mg/dl vs 56.72 +/- 11.46 mg/dl; p < 0.05) were significantly higher in patients with GHD. Carotid artery intima-media thickness (IMT) values were significantly higher in patients with GHD compared to healthy children (IMT-L 0.53 +/- 0.058 mm vs 0.41 +/- 0.045 mm; IMT-R 0.54 +/- 0.048 vs 0.42 +/- 0.42 mm, respectively). CONCLUSIONS: No significant differences in parameters of cardiac systolic function and left ventricular mass were found between patients with GHD and healthy children. However, significantly higher Isovolumetric relaxation time (IVRT) values in the group of patients may suggest impaired diastolic function. Significantly elevated IMT values in the common carotid artery were observed in patients with GHD in comparison with healthy children, which may prove the onset of atheromatosis. PMID- 15526720 TI - Low bone mass in prepubertal children with thalassemia major: insights into the pathogenesis of low bone mass in thalassemia. AB - OBJECTIVE: Low bone mass occurs frequently in the aging thalassemic population. However, limited information exists on bone mass in children with thalassemia major (TM) during their first decade of life. STUDY DESIGN: Spinal bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DEXA) in 18 children (age 5.8 +/- 1.5 yr; M:F 8:10) with TM on hypertransfusion and iron chelation therapy. Serial BMD measurements were available for 11 of the 18 children. RESULTS: Weight and height z scores were 0.81 +/- 4.2 and -0.47 +/- 1.7 respectively. At the first BMD, four (22.2%) patients presented with BMD z scores less than -2.5, seven (38.8%) had BMD z scores between -1 and -2.5, while the remaining seven (38.8%) had normal BMDs (z score above -1). The mean decline of BMD z score was -0.38/year (p = ns). BMD z scores correlated with height z scores (p = 0.039), but not with liver enzymes, serum ferritin levels, or thalassemia genotypes. CONCLUSIONS: Low bone mass is present in most children with TM despite hypertransfusion and optimal chelation, adequate growth and lack of endocrine complications. PMID- 15526721 TI - Prevalence of microalbuminuria in young patients with type 1 and type 2 diabetes mellitus. AB - This study was designed to determine the prevalence of microalbuminuria and the associated risk factors in patients with childhood-onset diabetes mellitus (DM). One hundred and sixty-three patients (141 with type 1 DM [DM1] and 22 with type 2 DM [DM21), aged 8 to 28 years, were evaluated for albumin excretion rate and HbA(1c). The mean duration of DM was 8.1 +/- 3.4 and 5.5 +/- 3.9 years in DM1 and DM2, respectively. Persistent microalbuminuria and macroalbuminuria were observed in 11.3% and 2.8% of patients with DM1, and 18.2% and 4.5% in patients with DM2, respectively. In DM1, the duration of DM, age of onset, and HbA(1c) levels were significant predictors of microalbuminuria. Our observations suggest that screening for microalbuminuria should be started from early adolescence in patients with DM1 and DM2. PMID- 15526722 TI - Efficacy and safety of Valtropin in the treatment of short stature in girls with Turner's syndrome. AB - Valtropin (somatropin, BioPartners and LG Life Sciences [LGLS]) is a recombinant human growth hormone (GH) preparation produced using a yeast expression system. An open single-arm phase III study was conducted to evaluate efficacy and safety at a dose of 0.16 IU/kg/day (0.053 mg/kg/day) s.c. for 12 months in the treatment of short stature in girls (n = 30, aged 2-9 years) with Turner's syndrome. The primary efficacy variable was height velocity (HV) at 12 months. Secondary efficacy variables included serum GH dependent growth factors. HV increased from 3.8 +/- 1.8 cm/yr at baseline to 9.7 +/- 1.6 cm/yr (mean +/- SD) after 12 months of treatment. Marked treatment effects were also observed on other growth parameters, serum insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3). Treatment was well tolerated with no significant adverse events. It is concluded that Valtropin is as safe and effective as other human GH preparations for the treatment of growth failure in girls with Turner's syndrome. PMID- 15526723 TI - Thyroid hormone levels and their relationship to survival in children with bacterial sepsis and septic shock. AB - OBJECTIVES: Reported studies have showed alternations of thyroid hormones in critical illness mostly in adults and some in children. In this study, we aimed to measure thyroid hormone levels in children with sepsis and septic shock and investigate the relationship of these hormones with clinical state and survival. PATIENTS AND METHODS: Thyroid hormone levels of children with sepsis and septic shock, and age- and sex-matched controls were measured. RESULTS: There were 51 children in sepsis (group S), 21 children in septic shock (group SS) and 30 in the control (group C) group. Total triiodothyronine (TT3) levels were (nmol/l): 0.91 +/- 0.22, 0.64 +/- 0.23, 2.11 +/- 0.59; free triiodothyronine (FT3) (pmol/l): 0.027 +/- 0.006, 0.018 +/- 0.007, 0.049 +/- 0.010; total thyroxine (TT4) (nmol/l): 100.62 +/- 21.93, 65.79 +/- 19.35, 109.65 +/- 19.35; free thyroxine (FT4) (pmol/l): 18.06 +/- 3.87, 10.32 +/- 1.29, 19.35 +/- 3.87; and thyroid stimulating hormone (TSH) (mIU/ml): 5.0 +/- 2.0, 4.8 +/- 2.4, 5.2 +/- 3.0, in children with sepsis, septic shock, and controls, respectively. The TT3, FT3, TT4, and FT4 levels of group SS were significantly lower than those of groups S and C. The TT3 and FT3 levels of group S were lower than in group C, but there was no significant difference between TT4, and FT4 levels of groups S and C. TSH levels were slightly decreased in both sepsis and septic shock, but the difference was not significant. Eleven (21.6%) children with sepsis and 15 (71.4%) children with septic shock died (p < 0.001). The levels of TT3, FT3, TT4 and FT4 were markedly lower in non-survivors of groups S and SS compared to survivors (p < 0.001). CONCLUSIONS: These changes in the hypothalamo-pituitary thyroidal axis may suggest a possible prognostic value of thyroid hormone levels in children with sepsis and septic shock. To the best of our knowledge, this report is the first study to compare thyroid hormone levels in a large number of patients with sepsis and septic shock with those in healthy controls in childhood. PMID- 15526724 TI - The relationship between iron status and thyroid hormones in adolescents living in an iodine deficient area. AB - BACKGROUND: Iron may be only one of many nutritional factors that influence thyroid hormone metabolism in iodine-deficient areas. OBJECTIVE: To evaluate the effect of iron status on thyroid hormone profile in adolescents living in a mild iodine-deficient area in Turkey. SUBJECTS: Three hundred and thirty school-aged children with mean age 14 years were examined. METHODS: Free triiodothyronine, free tetraiodothyronine and thyrotropin levels were analyzed by chemiluminescent enzyme immunoassay. Hemoglobin and mean corpuscular volume were measured with a Hemocounter. Iron and total iron binding capacity concentration were determined by colorimetric photometric methods. Ferritin level was determined by immunoturbidimetry. RESULTS: Thyroid hormone levels of the children with anemia were not significantly different compared to children without anemia. No significant correlation was found between thyroid hormones and iron status. CONCLUSION: The thyroid hormone profile was not significantly affected in this area. This survey showed no correlation between iron status and thyroid hormone levels. PMID- 15526725 TI - Galactocele in a male infant with congenital hypopituitarism. AB - Galactocele is an uncommon benign breast lesion usually occurring in young lactating women. It is a rare cause of breast enlargement in children. We report a galactocele in a male child with congenital hypopituitarism and propose an association between the two conditions. PMID- 15526726 TI - Infantile osteopetrosis in four Thai infants. AB - Four Thai infants, aged between 4 and 23 months, had progressive abdominal distension, pallor and delayed or regressed developmental milestones, with age at onset of 1 month, 3 months, 4 months and 1 month, respectively. Clinical findings consisted of growth and developmental retardation, anemia, frontal bossing, marked hepatosplenomegaly, and hearing and visual impairment. Laboratory findings revealed moderate anemia, leukocytosis and thrombocytopenia. The radiographic findings comprised generalized sclerosis of all bones, including the cranial base, and obliteration of the medullary canals and trabecular patterns. The first and second patients, who had swelling of the wrist joints and prominent costochondral junctions, had hypophosphatemia, elevated levels of serum alkaline phosphatase, and metaphyseal flaring on their radiographs, which was consistent with infantile osteopetrosis complicated by rickets. After Stoss therapy, there were biochemical and radiological responses suggesting vitamin D deficiency in the first patient, but not in the second. The third patient, who had hypocalcemia, hypophosphatemia and normal levels of serum alkaline phosphatase, received vitamin D at 3000 units per day, without improvement. Despite frequent blood transfusions, all patients continued to deteriorate and were finally lost to follow-up. Rickets should be identified and treated at the onset, because treatment of rickets leads to improvement in well-being and an adequate clinical response to bone marrow transplantation. PMID- 15526727 TI - Wolfram (DIDMOAD) syndrome: report of two patients. AB - We report a girl with Wolfram syndrome who presented with juvenile-onset diabetes mellitus when she was 4 3/12 years old. Optic atrophy and high frequency sensorineural hearing loss were found at 7 and 9 5/12 years of age, respectively. Her younger brother also developed Wolfram syndrome when he was 3 2/12 years old. Wolfram syndrome is also called DIDMOAD (diabetes insipidus, diabetes mellitus, optic atrophy and deafness). This syndrome is transmitted as an autosomal recessive trait and is a progressive neurodegenerative disorder. It should be considered in a diabetic patient with unexplained optic atrophy, hearing loss, or polyuria and polydipsia in the presence of adequate blood glucose control. Visual acuity should be checked annually in patients with juvenile-onset diabetes mellitus. Optic atrophy should be considered if visual acuity is impaired. PMID- 15526728 TI - Persistent hyperinsulinemic hypoglycemia presenting with a rare complication: West syndrome. AB - BACKGROUND: Persistent hyperinsulinemic hypoglycemia of infancy (PHHI) is characterized by disproportional secretion of insulin from pancreatic beta-cells. Although one of the manifestations of hypoglycemia is West syndrome, it is rarely reported in PHHI. PATIENT REPORT: A 6 month-old girl who was followed up with the diagnosis of PHHI was admitted to hospital with the complaint of jerky movements at her extremities. EEG revealed the typical pattern of hypsarrhythmia leading to the diagnosis of West syndrome. CONCLUSION: To our knowledge, there is only one report in the literature of West syndrome as a manifestation of PHHI, and that was the hyperammoniemic form of the disease. The present report is the first of normoammoniemic PHHI leading to West syndrome. We wish to highlight the potential risks of PHHI, especially in inadequately treated patients, and to emphasize that close neurological follow-up is very important in children who suffer from PHHI. PMID- 15526729 TI - A survey of personality disorders. PMID- 15526730 TI - Thrombophlebitis: a common complication of amiodarone. PMID- 15526731 TI - Definitive statements should only be made with evidence. PMID- 15526732 TI - Suggestions for performing aspiration of breast cysts. PMID- 15526733 TI - NSAIDs should not be used in treatment of stress fractures. PMID- 15526734 TI - Teenage pregnancy prevention: what can we do? PMID- 15526735 TI - Diagnosis and treatment of basal cell and squamous cell carcinomas. AB - Rates of squamous cell and basal cell carcinomas have been increasing, possibly as a result of increased exposure to ultraviolet radiation. Primary care physicians can expect to diagnose six to seven cases of basal cell carcinoma and one to two cases of squamous cell carcinoma each year. Basal cell carcinomas may be plaque-like or nodular with a waxy, translucent appearance, often with ulceration and telangiectasia. They rarely metastasize and are treated with excision, cryotherapy, electrodesiccation and cautery, imiquimod, 5-fluorouracil, or photodynamic therapy (the latter is not approved for this purpose by the U.S. Food and Drug Administration), although surgery results in the fewest recurrences. Actinic keratoses are scaly keratotic patches that often are more easily felt than seen. They are amenable to any of the destructive techniques described above, with the exception of photodynamic therapy. Squamous cell carcinomas arise from keratotic patches and become more nodular and erythematous with growth, sometimes including keratin plugs, horns, or ulceration. Because they may metastasize, they often are treated with excisional biopsy. PMID- 15526736 TI - Pneumonia in older residents of long-term care facilities. AB - Compared with community-dwelling persons, residents in long-term care facilities have more functional disabilities and underlying medical illnesses and are at increased risk of acquiring infectious diseases. Pneumonia is the leading cause of morbidity and mortality in this group. Risk factors include unwitnessed aspiration, sedative medication, and comorbidity. Recognition may be delayed because, in this population, pneumonia often presents without fever, cough, or dyspnea. Accurate identification of the etiologic agent is hampered because most patients cannot produce a suitable sputum specimen. It is difficult to distinguish colonization from infection. Colonization by Staphylococcus aureus and gram-negative organisms can result from aspiration of oral or gastric contents, which could lead to pneumonia. Aspiration of gastric contents also can produce aspiration pneumonitis. This condition is not infectious initially and may resolve without antibiotics. Antibiotics for the treatment of pneumonia should cover Streptococcus pneumoniae, Haemophilus influenzae, gram-negative rods, and S. aureus. Acceptable choices include quinolones or an extended spectrum beta-lactam plus a macrolide. Treatment should last 10 to 14 days. Pneumonia is associated with significant mortality for up to two years. Dementia is related independently to the death rate within the first week after pneumonia, regardless of treatment. Prevention strategies include vaccination against S. pneumoniae and influenza on admission to the care facility. This article focuses on recent recommendations for the recognition of respiratory symptoms and criteria for the designation of probable pneumonia, and provides a guide to hospitalization, antibiotic use, and prevention. PMID- 15526737 TI - Assessment and management of personality disorders. AB - Patients with personality disorders are common in primary care settings; caring for them can be difficult and frustrating. The characteristics of these patients' personalities tend to elicit strong feelings in physicians, lead to the development of problematic physician-patient relationships, and complicate the task of diagnosing and managing medical and psychiatric disorders. These chronic, inflexible styles of perceiving oneself and interacting with others vary widely in presentation. In the Diagnostic and Statistical Manual of Mental Disorders, 4th ed., these styles are categorized into three clusters based on their prominent characteristics: cluster A, the odd or eccentric (e.g., paranoid, schizoid, schizotypal); cluster B, the dramatic, emotional, or erratic (e.g., antisocial, borderline, histrionic, narcissistic); and cluster C, the anxious or fearful (e.g., avoidant, dependent, obsessive-compulsive). Knowledge of the core characteristics of these disorders allows physicians to recognize, diagnose, and treat affected patients. The goal of management is to develop a working relationship with patients to help them receive the best possible care despite their chronic difficulties in interacting with physicians and the health care system. Effective interpersonal management strategies exist for these patients. These strategies vary depending on the specific diagnosis, and include interventions such as the use of specific communication styles, the establishment of clear boundaries, limit setting on the patients' behavior and use of medical resources, and provision of reassurance when appropriate. Additionally, medications may be useful in treating specific symptoms in some patients. PMID- 15526738 TI - Pregnancy prevention in adolescents. AB - Although the pregnancy rate in adolescents has declined steadily in the past 10 years, it remains a major public health problem with lasting repercussions for the teenage mothers, their infants and families, and society as a whole. Successful strategies to prevent adolescent pregnancy include community programs to improve social development, responsible sexual behavior education, and improved contraceptive counseling and delivery. Many of these strategies are implemented at the family and community level. The family physician plays a key role by engaging adolescent patients in confidential, open, and nonthreatening discussions of reproductive health, responsible sexual behavior (including condom use to prevent sexually transmitted diseases), and contraceptive use (including the use of emergency contraception). This dialogue should begin before initial sexual activity and continue throughout the adolescent years. PMID- 15526740 TI - Evaluating the patient with an ankle or foot injury. PMID- 15526741 TI - Cutaneous ulceration in a patient with HIV. PMID- 15526743 TI - Information from your family doctor. Blood pressure monitors. PMID- 15526742 TI - Information from your family doctor. High blood pressure. PMID- 15526744 TI - [125 years of infant mortality registration in Croatia (1874-1999)]. AB - Infant mortality data in Croatia have been registered in the last 125 years. A tremendous decline has happened during this period from 295.2/1000 live births in 1874 to 7.7/1000 live births in 1999. A tenfold decline was obtained during the second half of the 20th century. In the last years the decline has been slower. Significant differences are found between the counties and also in the counties between different years. Further analysis is needed to make conclusions because the small number of live births influences a lot the infant mortality. During the past period the causes of infant mortality have changed, too. There were classical reasons such as inadequate care about children, low hygienic conditions, poor nutrition, and infectious diseases. Today, causes of infant mortality in Croatia don't differ from those in developed countries. In the first place are perinatal causes and congenital malformations. The neonatal mortality accounts for 75% of infant mortality. Having in mind current reasons of infant deaths, it is not easy to separate the influences of socio-economical, biological or medical factors. It is particularly impossible to predict the influence of a risk factor on an individual case, because the outcome under the same conditions could be quite different. PMID- 15526745 TI - [Smoking habits among health staff in the county of Koprivnica-Krizevci- comparative study 1998 and 2002]. AB - The study examined smoking habits among health care workers of all health institutions in the County of Koprivnica-Krizevci with the aim of establishing the prevalence. The first research of smoking habits was done in 1998 in order to get basic data about smoking habits, and the second research was carried out in 2002 with the aim of establishing a trend of smoking habits. An anonymous qestionnaire was given to all employees of all health institutions in the County. It was answered by 508 persons (44%) in 1998 and by 622 persons (50%) in 2002. As far as profession is concerned, 49 (53)% of those who answered the questionnaire were doctors, 24 (42)% dentists, 7 (9)% pharmacists, 53 (58)% were professional nurses and 28 (37)% administrative-technical non-health professionals. The data in parentheses are for 2002. The survey showed that of all employees who answered the questionnaire 34% in 1998 were smokers and 31% in 2002, while 17% were ex smokers (1998 and 2002) and 48% in 1998 and 52% in 2002 have never been smokers. Although the damage of smoking and consequences is a matter of every day talk about, and the employees of health institutions are the first expected to set an example to become aware and informed, the data of this research showed that the decrease of prevalence of smokers among health care workers is very slow. PMID- 15526746 TI - [Impact of posttraumatic stress disorder symptoms and their intensity on patients working ability]. AB - After the war in Croatia, many people are suffering from posttraumatic stress disorder (PTSD). The aim of this research was to analyse the impact of symptoms of PTSD and their intensity on working ability of those patients. Two groups of patients with PTSD hospitalized at Psychiatric Department of osijek Clinical Hospital were interviewed. The first one consisted of 80 patients who were (by psychiatrist) estimated as able to work, and the second of 80 patients who were estimated as unable to work. In this research we used ICD-10 classification for diagnosis of PTSD, self-estimate scale for evaluation of intensity of simptoms of PTSD, Brief psychiatric ratin scale (BPRS) and medical files. Data were statistically arranged by SAS. Working status is correlated to values on Brief psychiatric rating scale (BPRS) and self-estimate scale for evaluation of symptoms intensity of PTSD. Work disability was statistically significantly correlated to: earlier appearance of symptoms of PTSD, more frequent reminding of stressor by live memories, and symptoms intensity. The intensity of symptoms was statistically more significantly correlated to work ability than their form, and the number of stressors lived through the war (through more severe illness) had more influence on work ability than characteristics or durability of a stressor. PMID- 15526747 TI - [Reconstruction of nasal tip in combined technique of cartilage disc grafts and onlay graft]. AB - Rhinoplasty is often thought of as the most challenging of all aesthetic procedures. Irregularities of nasal tip implicate decreasing of nasal function and are generally regarded as an unattractive facial feature. The combined operative techniques which required extracorporeal septoplasty, incorporation of cartilage disc graft and onlay graft were performed in nineteen patients. Among the nineteen patients, fifteen had procedure of primary rhinoplasty and in four of them secondary or revised rhinoplasty was made. Indications for this operative technique were in patients with boxy nasal tip, bifid nasal tip, lateral alar tethering with extremely fat or thin nose skin. We emphasize that this technique is very successful with an acceptable percentage of postoperative complications. The tecnique is method of choice for reconstruction of extreme nasal tip irregularities where the usual reconstruction technique does not give satisfactory results. PMID- 15526748 TI - [Specific anesthetic management in laparoscopic surgery]. AB - Laparoscopic surgery is a technique by which abdominal surgical treatments are performed in minimally invasive way. Many operations that once required long hospitalization are now being performed with laparoscopic technique. It has significantly reduced postoperative pain and the number of complications, shorter hospital stay, with faster recovery, and more rapid return to normal activities. These are only some of the advantages of laparoscopic surgery. The first laparoscopic cholecystectomy in Croatia was performed on May 14, 1992, at the University Department of Surgery of "Sveti Duh" General Hospital in Zagreb. By the introduction of this method as the "gold standard" in the treatment of cholelithiasis, laparoscopic surgery has been widely accepted, and today, ten years later, has become a recognized and routine surgical method. Because of its particular performance, laparoscopic surgery requires some specific anesthesiologic adjustments. Surgical treatments are usually performed in general endotracheal anesthesia, although regional or local one could be applied too. The choice of anestesiologic technique and anesthetics depends upon preoperative patient's condition, the kind of surgical treatment, anesthesiologist's evaluation, and at last upon the cooperation among patients, anesthesiologist and surgeon. The basic aim of anesthesia is respiratory and hemodynamic patient's stability, and the choice of anesthesiologic technique and anesthesia should enable satisfactory analgesia, amnesia, muscular relaxation, faster recovery and shorter hospital stay. PMID- 15526749 TI - [Contemporary surgical treatment of sellar region tumors]. AB - Due to improved diagnostics, and primarily neuroradiological methods, an increased incidence of sellar region tumors is marked, among which the most common are pituitary adenomas. Sellar region tumors cause dysfunctions of the hypothalamo-pituitary axis as well as surrounding neurovascular structures. Despite the development of pharmacological and radiation therapy, surgical treatment is in these cases still a dominant therapeutical method. The author displays an overview of contemporary surgical methods for the treatment of sellar region tumors, as well as an overview of other adjuvant methods, especially pharmacological. Described are the results of over 1200 patients surgically treated during the past 20 years for sellar region tumors, as well as their complications and adverse effects. The author argues for respecting modem consensus on standardization of results of treating sellar region tumors as well as the consensus on professional and technical equipment of doctors and institutions that deal with this complex work. PMID- 15526750 TI - [General psychosomatics in children and adolescents]. AB - The purpose of this study is a review of contemporary knowledge from the field of children and adolescent psychosomatics by using standard textbooks and journals. Data are chosen according to their importance and easy reference for a clinical study and scientific research. Psychosomatics can be defined as medicine of mutual relationships in the widest sense. It seems that factors of bio-psycho social area participate in etiology and pathogenesis, i.e. complex genetic, developmental, psychodynamic, neuroanatomical, psychoneurophysiological, neurobiochemical, psychoneuroimmunological, psychoneuroendocrinal, familiar, social and probably other factors. Children and adolescent psychiatry, pediatrics are mutually complementary. In the medical-psychosomatic field there is consideration in the form of spatial-operative models that have to be amended through the concepts of unconscious and preconscious as well as through a temporal-systematic perspective. A team approach will be applied in a therapy. PMID- 15526751 TI - [Transfusion-associated graft-versus-host disease]. AB - Transfusion-associated graft-versus-host disease (TA-GVHD) is rare but usually fatal complication of transfusion of any blood component containing viable T lymphocytes. TA-GVHD manifests as an acute syndrome characterized by the dysfunction of the skin, liver, gastrointestinal tract and bone marrow. The development of bone marrow aplasia increases the risk for hemorrhage and infection and most patients die within 1 month of transfusion. The risk associated with an individual transfusion depends on the number and viability of contaminated lymphocytes, the susceptibility of the patient's immune system to their engraftment and the degree of disparity in histocompatibility antigens between the donor and the patient. The risk of TA-GVHD are highest in recipients with immunodeficiency or immunosuppression. Prevention of TA-GVHD is of great importance as it cannot be treated successfully. Patients at risk must be identified and transfused with irradiated cellular blood products, as gamma irradiation inhibits the proliferation of donor lymphocytes and is currently the only recommended method for TA-GVHD prevention. PMID- 15526752 TI - [The role of finasteride in prostate cancer]. PMID- 15526753 TI - The role of MIP-1alpha in experimental hypersensitivity pneumonitis. AB - S. rectivirgula (SR) causes Farmer's Lung Disease, a classic example of hypersensitivity pneumonitis (HP). We utilized a model of experimental hypersensitivity pneumonitis (EHP), antibody to MIP-1alpha and MIP-1alpha -/- mice, to test the hypothesis that MIP-1alpha is essential in the development of EHP. Treatment of C57BI/6 mice with anti-MIP-1alpha antibody did not change the extent of pulmonary histology abnormalities, BALF cell number or characteristics, or BALF concentration of IL12p40, TNF, IL1alpha and IL6, after an i.t. challenge with SR. MIP-1alpha -/- animals responded similarly to wild-type (wt) animals in the extent and nature of pulmonary histologic changes and BALF cell number and type after a single i.t. injection of SR There was a dose-response relationship between the amount of SR and BALF IL12p40, MCP-1 and IL6 in both strains, and MIP 1alpha in wild-type animals. We next transferred SR cultured spleen cells from SR sensitized mice (both wt and MIP-1alpha -/-) to naive recipients. Lung histology and BALF characteristics after SR i.t. challenge of the recipients were used to determine if adoptive transfer had occurred. Cultured cells from MIP-1alpha -/- animals were fully capable of transferring EHP to recipients. There was no difference of BALF TNF, IL6 and IL1alpha between the strains, but there was more MCP-1 and IL12p40 in the MIP-1alpha -/- mice than in the control mice. MIP-1alpha is not necessary for the recruitment of cells into the lung and BALF after i.t. administration of SR, or the development of cells able to adoptively transfer EHP. PMID- 15526754 TI - Signaling pathways in NSCLC as a predictor of outcome and response to therapy. AB - The 5-year survival rate for patients with stage III non-small cell lung cancer (NSCLC) is 10%. A number of genetic alterations are associated with this disease including mutations and amplifications of EGFR (70%) and Ras (20-30%), both of which are upstream of PI3K. Our previous data show that these regulate tumor radiation sensitivity. Here we ask whether the activation of this pathway has prognostic relevance in NSCLC. Two series of patients were retrospectively analyzed. The first series consisted of 23 Stage III NSCLC patients treated preoperatively with a chemo/radiation protocol. The second consisted of 12 Stage III NSCLC patients treated with chemo/ radiation without surgery who had survived more than 2 years. Expression levels of EGFR and Her-2 were assessed by immunohistochemical staining. PI3K signaling was evaluated by staining for phosphorylated Akt (P-Akt), a downstream target of PI3K. The staining for EGFR, Her-2, and P-Akt were related to outcome in the two groups. Additionally, the importance of PI3K signaling was evaluated in 3 NSCLC cell lines using a pharmacological blockade of PI3K by LY294002. In the first series of patients, 43% were positive for EGFR, 5% for Her-2, and 82.6% for P-Akt. Of the survivors, 25% were positive for EGFR, 0% for Her-2, and 42% for P-Akt. For P-Akt, this difference had a probability calculation of 0.003. The three NSCLC cell lines that we tested were found to have high levels of P-Akt. Pharmacologically inhibiting PI3K led to decreased Akt phosphorylation and radio sensitization of all three cell lines. The finding that NSCLC survivors treated by radiation have lower levels of PI3K and Akt signaling is consistent with the idea that inhibition of Akt leads to radio sensitization. This further suggests that Akt might be a useful target for sensitization of NSCLC to radiation. PMID- 15526755 TI - Lung inflammatory responses and hyperinflation induced by an intratracheal exposure to lipopolysaccharide in rats. AB - Exposure of the respiratory tract to lipopolysaccharide (LPS) induces acute local inflammation and tissue injury associated with the various deliveries of LPS. To determine potential association of local inflammatory responses with respiratory tract dysfunction, infiltration of inflammatory cells, production of inflammatory mediators, lung hyperinflation and edema were measured in Wister rats 2, 4, and 24 h after an intratracheal administration of LPS at different doses (5, 50, 500 and 5000 microg/ml/kg). Lung hyperinflation determined by an increased excised lung gas volume was significantly increased 2 and 4 h after LPS instillation and lung edema occurred from 2 h onward. Peak BAL levels of TNFalpha appeared at 2 h, MCP-1 at 4 h, and IL-6 at 2 and 4 h, while BAL levels of IL-1beta were increased during 24 h after the intratracheal instillation of LPS. Neutrophilia in BAL fluid was noted from 2 h post-challenge. Our results demonstrate a clear dose related change in the lung weight at 4 and 24 h, in the BAL levels of MCP-1 at 4 h, and IL-6 and IL-1beta at 2 and 4 h. It seems important to understand polymorphisms of LPS-induced lung hyperinflation and inflammation. Lung hyperinflation and inflammation may be independent during the development of acute lung injury. PMID- 15526756 TI - Osteopontin is strongly expressed by alveolar macrophages in the lungs of acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) is characterized by an intense inflammatory response in the lung parenchyma. Recent studies suggest that excessive nitric oxide (NO) production mediated by inducible NO synthase (iNOS) in macrophages is partially involved in mediating acute lung injury in ARDS. On the other hand, osteopontin (OPN) is a cytokine which is capable of inhibiting NO production by suppressing iNOS mRNA expression in macrophages. In this study, we investigated the expression of OPN in the lungs of 10 patients with ARDS. In most patients, OPN is strongly expressed on alveolar macrophages. In addition, we produced a murine model for ARDS by intratracheal administration of lipopolysaccharide and investigated the expression of endogenous OPN and iNOS in the lungs of ARDS mice. Immunostaining demonstrated that in vivo OPN protein was coinduced with iNOS protein predominantly in the accumulating alveolar macrophages. OPN mRNA expression was also coinduced with iNOS mRNA, but was induced more slowly than iNOS mRNA in the lungs of ARDS mice. These results suggested that OPN, which may reduce NO production of macrophages by inhibiting iNOS expression, is significantly induced and expressed on alveolar macrophages in the lungs of ARDS. It is possible that OPN is partially involved in playing a protective role against excessive production of NO in ARDS. PMID- 15526757 TI - Economic evaluation of a community-based pulmonary rehabilitation program for chronic obstructive pulmonary disease. AB - Pulmonary rehabilitation has been demonstrated to be efficacious in chronic obstructive pulmonary disease (COPD), however, its cost-effectiveness is largely unknown. The present study determined the cost-effectiveness of a community-based pulmonary rehabilitation program for COPD patients with mild, moderate, and severe disease. We compared the direct costs (in Canadian dollars) and disease specific quality of life (measured by the St. George's Respiratory Questionnaire, SGRQ) of patients with COPD (N = 210) who enrolled in the rehabilitation program in Edmonton, Canada one year before and after completion of the program. To determine temporal trends in health service utilization between 2000 and 2002 we used similar data from 592 COPD patients from the same region who did not participate in the rehabilitation program. We found that the health status of patients enrolled in the program improved significantly following pulmonary rehabilitation, irrespective of the severity of disease (total SGRQ score improved by 4.85%, p = 0.001). The total direct cost per 100 person-years of follow-up before the program was $122,071 (SE = 29,566); after the program it was $87,704 (SE = 26,146). The average reduction of total costs before and after the program was $34,367 per 100 person-years or approximately $344 per person per year (p = 0.02). Over one-year, pulmonary rehabilitation was associated with decreased health service utilization, reduced direct costs and improved health status of COPD patients. This suggests that pulmonary rehabilitation is cost effective for patients with relatively high utilization of emergency and hospital based services. PMID- 15526758 TI - [Hansen's disease-associated disabilities in sanatoria residents. A study of the actual situation in three sanatoria located in the Setouchi area]. AB - PURPOSE: Clarification of details of Hansen's disease in Japan is important for obtaining an adequate understanding of the present circumstances regarding control and for formulation of specific suggestions for future measures. The average age of residents in national Hansen's disease sanatoria reached 74.9 in May 2002 and most of these suffer from serious disabilities caused by the disease. In order to implement measures facilitating their return to society or secure living in sanatoria, a proper understanding of the residents' physical situation is essential. Therefore, the present study was conducted of residents suffering from Hansen's disease in Japan. METHOD: A direct interview was conducted with all 1282 residents of three sanatoria, located in the Setouchi area: Nagashima-aiseien, Oku-komyoen, and Oshima-seishoen. An investigator visited the rooms of the residents, interviewed them using a questionnaire, which consisted of about 100 items including the OPCS scale, and filled out an answer form. The reply rate was 64%. RESULTS: The average age of the residents was found to be 72.8 years, their average age when they entered the sanatoria was 26.0 years, and the average period of residence was 52.4 years. The damage they suffer as a result of the isolation policy is physical, mental, economical and social. That is to say, their entire life is harmed as a direct result of the Leprosy Prevention Law or the Eugenic Protection Law with discrimination in society and negative effects on their families. Their physical condition features serious disabilities in the use of fingers, as well as in walking and seeing caused by peripheral nerve damage. Such disabilities are related to their age and period of residence, and in addition, these in use of their fingers are related to the work which residents were forced to do in sanatoria. The results indicate that patients' physical condition is related to their age, how long they have had the disease and their work in sanatoria. CONCLUSION: It is clear that previous Hansen's disease control in Japan has caused a great deal of harm, from which it is very difficult to recover so that residents have to live with serious problems. In order to implement measures allowing their return to society or more secure life at sanatoria, sufficient care is needed to treat progressive disabilities caused by ageing and prevent deterioration caused by peripheral nerve damage. PMID- 15526759 TI - [Zinc intake from daily meals in preschool children and its influence on their physical growth]. AB - OBJECTIVE: The aims of this study were to investigate the influence of dietary zinc intake on physical growth and to clarify effects of dietary factors in preschool children. METHODS: Food consumption of 30 children aged 5-6 years (15 boys and 15 girls) was surveyed using a weighing dietary method for three consecutive days each in spring and autumn. Daily nutrient intakes were calculated using the Standard Tables of Food Composition in Japan Fifth Revised Edition, 2000. The heights and weights were also measured in each season. They were classified into low, middle and high zinc intake groups, and their physical growth and dietary factors were compared. RESULTS: The average zinc intake was 6.4 +/- 1.1 mg/day in all children, which met the demands of the Japanese RDA (6.0 mg/day). Classification was a low zinc intake group (5.4 +/- 0.5 mg/day), a middle group (6.2 +/- 0.3 mg/day), and a high group (7.7 +/- 0.6 mg/day). The lower zinc intake group showed lower heights and weights, but all children who participated demonstrated increase during the 5 months from spring to autumn. In particular, increase in height in the low zinc intake group was the highest among the three groups. Regarding each food item contribution for the zinc intake, rice (20.53%), meat (16.28%), milk (15.57%), eggs (7.45%), and soybeans (6.87%) were the highest 5 of 19 items, accounting for 66.7% of the total. Zinc intake from vegetable foods (55%) was more than that from animal foods (45%). Intake of total energy, nutrients, and some food groups positively associated with average zinc intake. However, energy intake from confectionery in the lowest zinc intake group was highest among the three groups. CONCLUSION: Estimation of zinc intake of Japanese children using a weighing dietary method has suggested that even a relatively low dietary value might not significantly affect the physical growth of children. Rice is the main source and reduction of zinc intake from regular meals might be related to high consumption of confectionery. PMID- 15526761 TI - [Health status and socioeconomic factors related to home remedy practices among elderly subjects living in a community in Okinawa]. AB - OBJECTIVE: The purpose of the present study was to clarify health status and socioeconomic characteristics of elderly persons who practice home remedies as complementary alternative medicine. Whether such usage is associated with health related factors was also assessed. METHODS: The subjects are 243 independent elderly living at home in A-village of Okinawa prefecture, aged 65 years or older. A self-administered questionnaire was mailed to all subjects in 2001. Included were questions on socioeconomic indicators, health status, IADL, health habits, social network, social support and home remedy practices. All data were statistically analyzed by Student's t test and the chi-square test. In addition logistic regression was used to calculate the odds ratios for current practitioners for each of the factors. RESULTS: The present study included 200 subjects who responded and completed all questions. (Table 2). The main results are as follows: The practitioner rate for home remedies was 50.8% in males and 71.9% in females, the difference being significant (P<0.05). Usage was found to be significantly related to hypertension (P<0.05), arthritis and rheumatic disease (P<0.01), some health habits and provision nursing of social support (OR = P<0.05). Logistic regression analysis adjusted for age, gender, and income revealed current practitioners of acupuncture to have a significantly higher level of history of hospital admission in the previous year (OR = 0.2 P<0.05), food control habits (OR = 4.3 P<0.05) and provision nursing of social support (OR = 4.2 P<0.05) than non-current practitioners. CONCLUSION: The findings of this study suggest that home remedy practices are associated with health-related habit and provision of social support for good quality of elderly life. The purpose is not only physical care but also health maintenance and promotion. PMID- 15526760 TI - [Relationships between finding of calcaneal quantitative ultrasound and lifestyle in Japanese college students]. AB - PURPOSE: The actual situation regarding the lifestyle of college students in Japan cannot be said to be healthy and future reduction of bone mass is a possible matter for concern. However, there have been only a few reports about the relationship between lifestyle during this period and bone mass, and especially none focusing on males not affected by female hormones. The purpose of this study was thus to investigate the relationship between bone mass and lifestyle in college students of both sexes. METHODS: Seven hundred and sixty-six college students of both genders were enrolled in this study as subjects and underwent calcaneal quantitative ultrasound with an AOS-100 device (ALOKA). At the same time physical factors, lifestyle and nutrition were also examined with laboratory findings. RESULTS: Physical factors such as stature, weight, body mass index (BMI), percentage body fat and gripping power were more strongly related to calcaneal quantitative ultrasound among females than in male students. Multiple regression analysis using calcaneal Osteo Sono-Assessment Index (OSI) as the dependent variable, and physical factors, lifestyle and nutrition, and laboratory findings as the independent variables, indicated that regular physical activity was strongly related to OSI in both sexes. The OSI in the alcohol-consuming group was significantly higher than that in the non alcohol-consuming group in males, and demonstrated a significant negative correlation with liver function markers, i.e. ALT (GPT) IU/l. CONCLUSION: It is very necessary to undertake daily physical activity for primary prevention of osteoporosis in both males and females. Bone mass in the alcohol-consuming group was here found to be significantly higher than that in the non alcohol-consuming group, but the study suggested that if the amount of alcohol consumed reaches an extent where liver function markers are affected, bone mass may decrease. PMID- 15526762 TI - [An attempt to prevent lifestyle-related diseases of schoolchildren in collaboration with parents, schools and communities]. AB - PURPOSE: The authors focused on preventing lifestyle-related diseases among schoolchildren on the basis of health surveys with blood examinations and questionnaires on lifestyle in collaboration with parents, YOGO teachers, and public health nurses. The present study purposed to evaluate this approach using the PRECEDE-PROCEED Model. METHOD: The authors inquired of schoolchildren's parents their awareness of their children's health through health surveys of their children. Additionally, changes in the role of YOGO teachers and public health nurses before and after our attempt were assessed. The significance and problems with our approach were then evaluated based on the PRECEDE-PROCEED Model. RESULTS: Based on the PRECEDE-PROCEED Model, it was clarified that, for prevention of lifestyle-related diseases among schoolchildren, health surveys with blood examinations and questionnaires on lifestyle increased parents' interest in their children's health. On the basis of the results of health examinations, the role of YOGO teachers changed from leading figure for general health education to supporters of health promotion for individuals, with provision of health education classes in schools for this purpose. The role of public health nurses also changed from providing health education at the request of schools to supporting families to improve their lifestyle, including that of their children. CONCLUSIONS: In collaboration with children's parents, YOGO teachers and public health nurses, the present approach to prevent lifestyle related diseases of schoolchildren on the basis of health surveys has advantages for increasing interest in schoolchildren's health, and should be useful for health promotion. PMID- 15526763 TI - [Factors related to municipality expectations of retaining physicians in public medical facilties]. AB - OBJECTIVES: This study was conducted to investigate the association between municipality expectations of retaining physicians in public medical facilities and their evaluation of the physicians from key aspects considered important for practicing community medicine. METHODS: A cross-sectional study using a self reported postal questionnaire survey was conducted with all 3,152 local governments in Japan from July to September, 2000. Three thousand and fifty-nine (94%) local governments responded. Of the responders, 1,315 (42%) operated public medical facilities. These local governments evaluated their expectations of retaining physicians in medical facilities and the physicians themselves focusing on the following aspects considered important for practicing community medicine: degree of government satisfaction with the physician's involvement in public health and welfare services; primary care for emergency patients; human relationships between the physician and the population, staff of the facility or the municipal officials; public estimation of the facility; income and expenditure of the facility; understanding of the local government's system. RESULTS: The subjects were 1,092 local governments (83.0%) that responded to all of the questions. The proportion of municipalities expecting to retain physicians was 56% overall, 61% in rural areas and 44% in urban areas (P<0.001). The proportion of governments satisfied with the physicians' human relationships was higher than the other factors and the proportion satisfied about income and expenditure of facilities was the lowest. Multiple logistic regression analysis revealed that the degree of local government expectation of retaining physicians was significantly associated with the physicians' involvement in public health and welfare services (Odds ratio (OR) 1.8; 95% confidential interval (95%CI) 1.3 2.5), their human relationship with the population and public estimation of facilities (OR 1.6; 95%CI 1.1-2.2), and income and expenditure (OR 1.3; 95%CI 1.01-1.8 in all, OR 1.7; 95%CI 1.7-2.4 in rural areas). Primary care for emergency patients demonstrated a significant association in rural areas (OR 1.6; 95%CI 1.1-2.3). CONCLUSIONS: Municipal expectation of retaining physicians in public medical facilities was significantly associated with factors related to the integration of medical, public health and welfare systems, human relationship with the population, public estimation and income and expenditure of the facilities overall. In rural areas, there was a closer association with income and expenditure and a significant association with emergency care. These findings suggest that the local governments' thought related with the expectation of retaining physicians are influenced by their different needs for medical services in community healthcare. PMID- 15526764 TI - [Problems of tuberculosis measure based on hospitalization and medical treatment conditions of registered patients in Mie prefecture]. PMID- 15526765 TI - [A nationwide survey of municipalities' policy for smoking control and its implementation in relation to its priority grading by public health officers]. AB - PURPOSE: The purpose of this study was to investigate the present situation regarding implementation of smoking control programs and to clarify relations with public health officers' attitudes toward smoking control measures. METHODS: A questionnaire form was mailed to the 3,207 local municipalities throughout Japan. The items included in the form were the contents of the smoking control programs, educational activities for passive smoking prevention, interaction with the media to spread education and awareness, and public health officers' attitudes toward smoking control measures. RESULTS: Of 3,207 municipalities, 2,570 (80.1%) responded. Over 95% of local municipalities were undertaking smoking control programs. Among the programs carried out in practice, dividing areas into smoking and non-smoking sections in local government office buildings had the highest proportion of support (80%). On the other hand, complete prohibition of smoking in local government office buildings and non-smoking support programs were conducted by less than 20%. The dissemination of education activities were executed in 60 percent of government office buildings, and 36% of schools, but only 20% of athletic gyms, hospitals, and other facilities. Sixty percent of public health officers recognized the importance of smoking control programs. A non-smoking support program and complete prohibition of smoking in government office buildings were more likely to be executed in for local municipalities with a higher priority for smoking control programs. No significant association was found between attitudes regarding program importance and the division of areas into smoking and non-smoking sections in government office buildings. CONCLUSIONS: Our results suggest that promotion of smoking control measures in local municipalities may be dependent on the public health officers' attitude toward the issues. High priority should be given to assisting public health officials and other key personnel in recognizing the importance of implementing smoking control measures for attaining better health. PMID- 15526766 TI - [Pathophysiochemistry of acute death: an approach to evidence-based assessment in forensic pathology]. AB - The essential social and academic task of legal medicine is to devote itself to a multidisciplinary approach to problems at the interface of medicine and law. It includes forensic medical activity, in which one of the social concerns is to investigate the fatal mechanisms, survival time and physical activity, especially in traumatic and unexpected sudden death, by means of forensic pathological procedures. To meet the social requirements through reliable interpretation of those issues, systematic practical investigations are necessary, establishing the evidence-based assessment in forensic pathology. For that purpose, an approach based on the pathophysiochemistry of fatal mechanisms may be useful to aid or support pathomorphological observations. Essential markers in forensic pathophysiochemistry are the indicators of systemic responses involving acute phase reaction to traumas, i.e., circulatory, respiratory and central nervous system (CNS) functions. A comprehensive study based on previous investigations is necessary to establish practical markers and to promote their use in routine forensic casework. In the present paper, reviewing the literature, our data in routine casework are summarized. Routine forensic casework at our institute includes biochemistry on automated analyzer systems, immunohistochemistry using commercial kits and molecular biology by means of RT-PCR: 1) blood and urine biochemistry in general, 2) oxymetry, 3) serum and pericardial myocardial markers (creatine kinase MB, troponin I and T), 4) serum pulmonary surfactants (SP-A and D), 5) other serum markers including C-reactive protein, neopterin, catecholamines, cortisol, erythropoietin and S-100 protein, 6) pericardial natriuretic peptides, 7) urinary myoglobin, 8) immunohistochemistry of a pulmonary surfactant (SP-A) in the lungs, ubiquitin, S-100 protein and ssDNA in the brain, and 9) RT-PCR for a pulmonary surfactant (SP-A) in the lungs, ischemia and hypoxia-related factors (hypoxia-inducible factor 1A, vascular endothelial growth factor and erythropoietin) in the brain, heart and kidneys. Further accumulation of practical data may be essentially important to establish evidence for medico-legal assessment in individual cases and to renew forensic pathology in response to potential social requirements. PMID- 15526767 TI - [Induction mechanism of shock: applying the etiology in judgment of the cause of death in forensic practice]. AB - In the field of forensic medicine, shock has been identified as a cause of death owing to various kinds of exogenous insults. The etiology and pathogenesis of shock cannot be explained well by the usual gross appearance in medicolegal autopsies, because it is now generally established that the shock is a functional reaction of the vascular system to bodily injury, and that several organs are secondarily impaired during shock. Thus it seemed to forensic pathologists that these morphological changes in several organs after shock did not reveal any significant differences among the causes of death. We approached to the induction mechanism of shock, and we investigated what etiology induced these morphological changes after shock in order to identify shock as the cause of death. It is now generally accepted that the kidney is a target organ of shock, so we mainly investigated the cause of kidney disorder in a case of burn shock and hemorrhagic shock. 1. Consequences of bacterial translocation (BT) in the shock. The concept of BT indicates that the beginning of shock is induced by the loss of gut barrier function and consequent translocation of bacteria. In general, impaired gut barrier function can be caused either during the shock period by decreased intestinal blood flow and reduced oxygen delivery, resulting during reperfusion in a stage of increased intestinal blood flow, or at a later stage again by reduced flow. A variety of physiological stresses, such as trauma, hemorrhage, thermal injury, surgical operation, various kinds of drags and mental stress, have been shown to cause failure of the gut mucosal barrier, with translocation of bacteria/endotoxin from the gastrointestinal into the mesenteric lymph nodes, and translocation into remote organs and systemic circulation. 2. Burn shock. We designed to evaluate the BT in a burn shock rat model (following 20% full thickness scald injury). The p38 MAPK pathway is an important stress-responsive signal molecule pathway, and it is responsible for the production and signal transduction of cytokines. This pathway is activated by the bacterial LPS or ischemia, so we examined the effects of FR167653, a specific inhibitor of p38 MAPK, on the development of renal failure after the burn-induced intestinal barrier damage. Our study demonstrated that viable bacteria reached the remote organs after burn by quantitative bacterial culture data and FR167653 blocked the burn-induced intestinal barrier damage, and the immunohistochemical data showed that FR167653 prevented the accumulation of polymorphonuclear leukocytes (PMNs) in the glomerular capillaries after burn, and blockaded the burn-induced renal failure by serum UN assay. FR167653 especially decreased the phosphorylation levels of p38 MAPK in the infant kidney after burn, and TNF-alpha and IL-1beta mRNA decreased through the p38 MAPK pathway. The above-mentioned facts do provide additional support for the hypothesis that postburn renal failure is mediated by endotoxin associated with the bacterial translocation, and we identified the pathophysiologic role of p38 MAPK pathway in the development of renal failure after the burn-induced intestinal barrier damage. 3. Hemorrhagic shock. We evaluated the role of endogenous TNF-alpha in the renal failure and gut bacterial translocation induced by mild hemorrhagic shock (16.7% bleeding of total body blood via a common carotid catheter without fluid resuscitation). FR167653, a potent inhibitor of TNF-alpha up regulation through p38 MAPK pathway, significantly inhibited these increases of TNF-alpha. Adding to this, our study demonstrated that FR167653 prevented renal failure, such as the infiltration of inflammatory cells and tubular cell necrosis after hemorrhage, and the intestinal barrier damage was also dramatically improved by FR167653 treatment. These results show that derived endogenous TNF-alpha plays a key role in renal failure through p38 MAPK activation during the early phase of mild hemorrhagic shock, including the possible participation of BT. According to these results, we hypothesized that the invading leukocytes induced these organs failures after hemorrhagic shock, so we examined the appearances of leukocytes by the immunohistochemical myeloperoxidase (MPO) staining (marker staining for PMNs). The incidences of PMNs in these organs after mild hemorrhagic shock increased significantly, and FR157653 prevented the appearance of PMNs. These results showed the possible effective role of the PMNs on the occurrence of organ failure caused by mild hemorrhagic shock. 4. Forensic practice. Six hundred and seven forensic autopsy cases in our department of forensic medicine during the past 11 years between 1992 and 2002 were analyzed with regard to the cause of death. Shock cases accounted for 18% of all forensic autopsy cases, and among them 65% of cases identified hemorrhagic shock as the cause of death. So we investigated what good grounds to clearly identify the cause of death induced by hemorrhagic shock. Our experimental hemorrhagic shock data showed PMNs activation and priming during hemorrhagic shock, and it might be closely related to BT and remote organ failure. Consequently, we used the MPO staining method, and we immunohistochemically investigated several organs of our practical autopsy cases to detect the appearance of PMNs as a marker of shock induction. We compared the hemorrhagic shock with other causes of death, such as blood loss, asphyxia, drawing and head injury (intracranial hemorrhage). In every organ, a significant appearance of PMNs was observed in the hemorrhagic shock compared to the other causes of death. Especially, the appearance of PMNs in the heart was clear than that of the other organs in the hemorrhagic shock cases. Therefore, detecting the appearance of PMNs as a marker of shock induction is a very useful and significant method forjudging the cause of death in forensic practice. PMID- 15526768 TI - [Haplotype analysis of single nucleotide polymorphisms of mitochondrial DNA]. AB - Mitochondrial DNA (mtDNA) mutations observed in both non-coding control and coding regions are being used widely to characterize human evolution and for identification. Several methodological approaches have been available for detection of single nucleotide polymorphisms (SNPs). SNPs of the non-coding region were mainly analyzed by means of direct sequencing. Especially, sequence analysis of D-loop is often the final resort in forensic and ancient personal identification. However, sequencing is relatively expensive and time-consuming. Furthermore, because the mtDNA molecule is a single linked unit, the statistical significance in forensic cases requires mtDNA matching in comparison with large sequence data. In this study, we developed a procedure of technique simultaneous typing to 11 SNPs and 2 tandem repeat sites in the non-coding and coding region by use of allele-specific amplification. In an analysis to 631 unrelated Japanese individuals the 180 haplotypes were defined in this study, and the level of haplotypic diversity and random mach probability was similar to that obtained by sequencing of the human hypervariable region 1 in mtDNA. These results show the usefulness of mtDNA haplotype analysis by the presented method for personal identification. PMID- 15526769 TI - [Case of positive identification by digital superimposed comparison between photograph of the thoracic vertebrae front and thorax roentgenograph]. AB - We reported an autopsy case in which an antemortem thorax roentgenograph and a postmortem photograph of thoracic vertebrae front were available for digital superimposed comparison of contour of the vertebral column and provided a positive identification by the characteristic osteophyte formation. In the elderly, the thorax roentgenograph is often stored in medical institution. Osteophyte formation of the vertebral column has individual features with the aging and formed characteristic profiles of the vertebral column. Photographing of a cadaver's thoracic vertebrae front after removing of the thoracic and abdominal organ should be carried out to make a material for future comparison examination in personal identification. PMID- 15526770 TI - [Basic methodology in forensic autopsy]. PMID- 15526771 TI - [The legal issue of the present occupational physician system in crisis management of health damage]. AB - In present day Japan, when a crisis like the Bhopal accident occurs, due to defects in the current of industrial health law, effective crisis management cannot be taken to minimize health damage in both workers and residents. The current law characterizes industrial health as a part of the welfare service provided by employers for employees. Nevertheless, the company should be liable for all failure and damage including health impairment as a primary party. Moreover, in an emergency crisis, unlike the case of reparations, it is not accepted to argue whether the company should take absolute liability or not. Accordingly, in such cases, we consider it more appropriate to apply the "Polluter Pays Principle" and the principle of "Liability without Fault" to the company's responsibility. By these rules, the company should mobilize their own professionals, who are experts of managing crises, such as occupational physicians and/or industrial health professionals to minimize health damages among citizens in general. The company should take such a social responsibility in a crisis when it is the primary responsible party to the crisis. PMID- 15526772 TI - [Health certificate problems at the place of employment]. PMID- 15526773 TI - [Study on the relative breakthrough time (RBT) of various respirator cartridges]. PMID- 15526774 TI - [Relationship between self-oriented perfectionism and depressive mood in the industrial society]. AB - The purpose of this study was to investigate the relationship between self oriented perfectionism and depressive mood in the industrial society. 380 workers in a construction company replied to a questionnaire, which consisted of a Beck depression inventory (BDI) and a multidimensional self-oriented perfectionism scale (MSPS). The results showed that concern over mistakes (CM) had a positive correlation with depressive mood in both younger and older age groups. Personal standard (PS) had a negative relation with depressive mood in younger age groups, but no statistical difference was seen in older age groups. We considered that the background for depressive mood differs in different age groups. PMID- 15526775 TI - [Interview study on autonomous chemical management system and the contribution of occupational health specialists in companies]. AB - Under the circumstance that autonomous risk management for chemicals is required in Japan, it is necessary to define fundamental steps for developing chemical management system that are applicable in various types of companies and to understand the effective contribution of chemical or occupational health specialists to the system. For the purposes, we conducted interviews with companies which have an advanced chemical management system in Japan. As the result, each company had a certain policy about detailedness level of collected hazard and exposure information, and also had an efficient risk management system to ensure workers' health in depending on the business type and situations. Moreover, it was commonly observed that the specialists played major roles in developing tools for risk assessment and control, and then business lines led execution of the risk management with their supports. Based on the interviews, we showed a hypothesis of basic steps in introducing autonomous chemical risk management system at the workplaces. It is necessary to verify the hypothesis and to develop a simple system that is applicable to middle or small size companies as the next step. PMID- 15526776 TI - Assessing swine thermal comfort by image analysis of postural behaviors. AB - Postural behavior is an integral response of animals to complex environmental factors. Huddling, nearly contacting one another on the side, and spreading are common postural behaviors of group-housed animals undergoing cold, comfortable, and warm/hot sensations, respectively. These postural patterns have been routinely used by animal caretakers to assess thermal comfort of the animals and to make according adjustment on the environmental settings or management schemes. This manual adjustment approach, however, has the inherent limitations of daily discontinuity and inconsistency between caretakers in interpretation of the animal comfort behavior. The goal of this project was to explore a novel, automated image analysis system that would assess the thermal comfort of swine and make proper environmental adjustments to enhance animal wellbeing and production efficiency. This paper describes the progress and on-going work toward the achievement of our proposed goal. The feasibility of classifying the thermal comfort state of young pigs by neural network (NN) analysis of their postural images was first examined. It included exploration of using certain feature selections of the postural behavioral images as the input to a three-layer NN that was trained to classify the corresponding thermal comfort state as being cold, comfortable, or warm. The image feature selections, a critical step for the classification, examined in this study included Fourier coefficient (FC), moment (M), perimeter and area (P&A), and combination of M and P&A of the processed binary postural images. The result was positive, with the combination of M and P&A as the input feature to the NN yielding the highest correct classification rate. Subsequent work included the development of hardware and computational algorithms that enable automatic image segmentation, motion detection, and the selection of the behavioral images suitable for use in the classification. Work is in progress to quantify the relationships of postural behavior and physiological responses of pigs using thermographs. The results are expected to facilitate objective training of NN, hence improving the accuracy of the postural image-based assessment of the thermal comfort state. Work is also in progress to implement the analysis and assessment algorithms into computer codes for real time application. PMID- 15526777 TI - Dynamic responses of cattle to thermal heat loads. AB - The focal point of this limited review is bioenergetic research conducted in the Biological Engineering Research Unit at the U.S. Meat Animal Research Center (MARC), using recently developed instrumentation and analytical techniques. The dynamics of observed thermoregulatory responses in cattle to thermal heat load challenges are explored, with an emphasis on physiological and behavioral parameters of body temperature, respiration rate, and feed intake. Observations of body temperature, especially tympanic temperature, have shown hot environments to cause phase shifts, increased amplitude, and increased means for diurnal rhythms. Fractal analysis of body temperature records obtained at 2- to 10-min intervals has been found to be robust for objectively differentiating among responses of cattle in cool to hot environments, and it indicates a stress threshold of approximately 25 degrees C (coincident with declining feed intake). Other analyses determined a 21 degrees C threshold for increased respiration rate. The reported observations and analyses provide further understanding of how and why the animals respond to environmental challenges, an understanding that is necessary for refining performance models and developing energetic and thermoregulatory models. The dynamic responses are discussed in the context of establishing criteria for proactive environmental management for cattle during hot weather, using heat waves as an example. PMID- 15526778 TI - Nutritional strategies for managing the heat-stressed dairy cow. AB - Heat stress results from the animal's inability to dissipate sufficient heat to maintain homeothermy. Environmental factors, including ambient temperature, radiant energy, relative humidity, and metabolic heat associated with maintenance and productive processes, contribute to heat stress. The focus of this article is to identify environmental and metabolic factors that contribute to excessive heat load, describe how disruption of homeothermy alters physiologic systems of the cow, and discuss nutritional modifications that help to maintain homeostasis or prevent nutrient deficiencies that result from heat stress. Changes in diet are needed during hot weather to maintain nutrient intake, increase dietary nutrient density, or to reestablish homeostasis. Formulation for adequate nutrient intake is challenging because of the competition between nutrient density and other needs for the cow, including energy density and adequate dietary fiber. Lower DMI during hot weather reduces nutrients available for absorption, and absorbed nutrients are used less efficiently. An excess of degradable dietary protein is undesirable because of energy costs to metabolize and excrete excess N as urea. Optimizing ruminally undegraded protein improves milk yield in hot climates. Mineral losses via sweating (primarily K) and changes in blood acid-base chemistry resulting from hyperventilation reduce blood bicarbonate and blood buffering capacity and increase urinary excretion of electrolytes. Theoretical heat production favors feed ingredients with a lower heat increment, such as concentrates and fats, whereas forages have a greater heat increment. Improved dietary energy density and the lower heat increment associated with the inclusion of dietary fat must be coupled with limitations to fat feeding to avoid ruminal and metabolic disorders. Numerous nutritional modifications are used for hot weather feeding; however, many need further investigation to achieve specific recommendations. PMID- 15526779 TI - Strategies for managing reproduction in the heat-stressed dairy cow. AB - Establishment and maintenance of pregnancy is difficult in lactating dairy cows exposed to heat stress because of reductions in estrous detection rate and the proportion of inseminated cows that maintain pregnancy. The most common approach to ameliorate heat stress in developed countries has been to alter the cow's environment through provision of shade, fans, sprinklers, and so on. Nonetheless, seasonal variation in reproductive function persists. Increased understanding of bovine reproductive function and its alteration by heat stress has led to additional strategies for reducing deleterious consequences of heat stress on reproduction. These include hormonally induced timed artificial insemination, which can reduce losses in reproductive efficiency caused by poor detection of estrus, and embryo transfer, which can increase pregnancy rate by allowing embryos to bypass the period when they are most sensitive to elevated temperature (i.e., in the first 1 to 2 d after breeding). Other efforts are directed toward developing methods to protect the embryo from harmful actions of elevated temperature. Approaches being studied include manipulation of embryonic synthesis of heat shock proteins and use of antioxidants to reduce free radical damage associated with heat stress. It may also be possible to reduce the magnitude of hyperthermia caused by heat stress. This might be possible physiologically, for example by feeding of agents that affect thermoregulatory systems, or genetically by selecting for specific traits conferring thermal resistance. Finally, the development of bovine somatotropin as a lactational promotant means that it may be possible to extend lactations beyond 305 d and voluntarily discontinue inseminations during periods of heat stress. PMID- 15526780 TI - Livestock nutrient management concerns: regulatory and legislative overview. AB - A greater focus on manure nutrient disposition from concentrated animal-feeding operations has developed from environmentalists, concerned citizens, and regulatory agencies. The establishment and enforcement of manure nutrient regulations will alter the future of livestock production. Proposed legislation and strategies may provide a false sense of security regarding environmental preservation or restoration and may impose monitoring and record keeping on the livestock operators. Existing regulations and proposed regulations and strategies are presented. Implications of legislation and proposed strategies, policies, and regulations are discussed. Livestock operations will need to comply with regulations to remain in business and to minimize environmental liability. PMID- 15526781 TI - Nutrient balance on Nebraska livestock confinement systems. AB - Managing the environmental risk associated with livestock production is a significant challenge. Nitrogen and phosphorus are commonly implicated as the sources of ground and surface water quality problems associated with livestock production. The degree of imbalance between these nutrient inputs and the managed nutrient outputs for a livestock operation defines the magnitude of potential environmental risk and provides insight as to the underlying causes of these challenges. A nitrogen and phosphorus balance was constructed for 33 Nebraska confinement livestock operations. Twenty-five and 17 of these operations experienced significant nitrogen and phosphorus imbalances, respectively (50% more nutrient inputs than outputs). Nutrient inputs on many livestock operations were observed to be two to four times greater than nutrient outputs as managed crop and livestock products. Size of the livestock operation and the degree of integration of livestock with a cropping operation provided only limited explanation of the variation in nutrient balance observed among the individual operations. Management options that contribute to a more favorable nutrient balance were also identified. Management decisions related to feeding program and exporting of manure nutrients to off-farm users were observed to have a substantial impact on the nutrient imbalance. For modern livestock production systems to successfully respond to nutrient-related environmental problems, management strategies must be implemented that address the commonly experienced imbalances of nitrogen and phosphorus. PMID- 15526782 TI - Animal grouping strategies, sources of variation, and economic factors affecting nutrient balance on dairy farms. AB - Increasing environmental concerns are forcing animal industries to reevaluate current feeding practices and their relationships to nutrient excretion. Previous modeling efforts have used simple budgets of nutrient flows through animals, assuming a constant productivity level. This assumption is not valid if animals are not in a steady state. A response model of dairy cow production to levels of net energy for lactation (NEL) and crude protein (CP) was derived from an abrupt threshold and plateau model of individuals. Monte Carlo techniques were used to simulate populations of cows fed diets of various NEL and CP concentrations, to derive the optimum allocation of NEL and CP, and to estimate how the optimum is affected by herd production potential, prices of inputs, and uncertainty of parameters. The simulation showed that a 25% increase in milk production reduced N excretion per kilogram of milk produced by 8%. Improved knowledge of the biology involved and feed composition can reduce N excretion by an additional 8%. Grouping strategies and number of groups used affect optimum allocation of nutrients. An optimum of six milking groups per production unit was derived from the simulation and would reduce N excretion by 8% compared to herds fed in one group. PMID- 15526783 TI - An approach to nutrient management on dairy farms. AB - In the European Union, groundwater should contain less than 50 mg of nitrate/L. Individual countries have developed alternative strategies for phosphorus (P). In The Netherlands, regulations based on P limited the amount of manure applied per hectare. A more balanced P supply to the land has been achieved by transport of manure from surplus to deficit regions. Costs of processing of manure to pellets appeared to be (too) high. In animal production experiments, lowering the P content of concentrates and mineral supplements reduced P losses without an adverse effect on production. In addition to the European guideline for nitrate, ammonia volatilization should be reduced by 50 to 70%. Management practices for reducing nitrogen (N) losses were studied with a farm model, developed at PR. A combination of a more efficient use of fertilizer N, restricted grazing, and a more balanced diet, and, to a lesser extent, higher milk production per cow resulted in considerable reductions in nitrate leaching. The application of slurry by injection diminishes the ammonia volatilization at farm level by almost 50%. This technique has become obligatory, and is only allowed during the growing season. Other techniques, like low emission housing and covering of slurry storage have relatively high costs. Starting in 1998, farmers have to keep a record of nutrients on a balance sheet. A tax will be imposed on surpluses of N and P. This new instrument replaces the regulations based on P. To further improve efficiency of use of N and P, farmers have the nutrient balance sheet available as an integrated management tool. Urea content in bulk milk has been introduced as a new indicator for the utilization of N in the diet. Also, fertilizer applications are improved. Furthermore, an experimental farm was set up to integrate all available expertise and analyze the resulting nutrient flows and farm performance. PMID- 15526784 TI - Managing nutrients across regions of the United States. AB - Nutrient balance in the ecosystem involves profitability of the agricultural enterprise and commitments to resource management to maintain quality of air, water, and land resources. Phosphorus and N are the two nutrients of major concern, and they behave differently in soils. Most P adheres strongly to soil particles and moves laterally with the soil during erosion processes, but with high concentrations more P remains in soluble forms and moves in the water fraction. Most N is soluble and moves laterally or downward with soil water. Soil scientists and agronomists have researched soil processes, plant nutrition, cropping systems, and water quality issues mainly on a field and farm level, but now the movement is to management and regulation of nonpoint problems on a watershed basis as proposed in the Clean Water Action Plan. The plan recognizes the vast diversity of soil parent materials and climates among geographic areas, even among and within watersheds, that determine crop adaptation and cropping systems, the role of states in regulatory processes, and the need for local citizens to have operational involvement. This process insures that nutrient management guidelines will be more site-specific and solutions can be focused on the direct problem. Directed efforts will be needed to educate local citizens, landowners, and caretakers of agricultural enterprises, and regulatory agencies. Several factors, including economic and social incentives for implementation must be considered along with the technologies available. The solutions are multidisciplinary, will require long-term research to accommodate climate variation, and should be associated with a strong commitment to education. Public funding will be needed to support the effort. PMID- 15526785 TI - Inflammation in the bovine female reproductive tract. AB - Inflammation of the reproductive tract of a cow occurs when the physical and functional barriers to contamination are breached or specific infection occurs. Commonly, contamination occurs at parturition and to a lesser extent at estrus. Uterine contamination following calving is common, but most healthy cows are able to clear the uterus of bacteria in the first 2 to 3 wk after calving. Persistent infections are more likely to be caused by Actinomyces pyogenes. Specific venereal infections tend to be more host-adapted and produce a lower grade inflammation. Nonspecific bacterial contamination of the endometrium generally induces a neutrophilic influx into the stratum compactum and uterine lumen. Neutrophils phagocytize bacteria with the aid of opsonins in the uterine fluid. Mast cells and eosinophils may also contribute to the inflammatory reaction, which may damage the surface epithelium and release vasoactive substances that allow leakage of serum antibodies into the uterine secretions. Specific antibodies of immunoglobulin (Ig) isotype A, M, G1, and G2 in uterine secretions have been described. In model species, the immune capability of the uterus is influenced by steroid hormones, especially estradiol, which increases secretory component and both IgA and IgG content in uterine secretions and increases the activity of antigen-presenting cells in the uterus. Similar cyclic fluctuations in immune components have been described for cows, including changes in the population of subsurface cytotoxic and helper T cells and changes in the expression of major histocompatibility II antigen on surface cells. PMID- 15526786 TI - Implications of cloning for breed improvement strategies: are traditional methods of animal improvement obsolete? AB - Can the optimum animal be defined? Will that definition change over time, by location, by market demand? First, assume what may be impossible, that the perfect animal can be defined or that only a limited number of definitions of "perfect" are needed. Then, can the "perfect" animal to match a definition be found? Suppose such an animal is found. Then the question to be answered before trying to clone as a method of genetic improvement becomes "Is the animal perfect because of phenotype or genotype?" In other words, the P = G + E problem exists, which requires traditional methods of genetic evaluation and testing to determine whether genotype (G) or random environmental (E) effects or a combination leads to the apparent perfection in the phenotype (P). For most traits, additive genetic variance accounts for 10 to 50% of total variance, a fraction denoted as heritability. With a simple model, the best prediction of genotypic value is to reduce the apparent phenotypic superiority by multiplying by heritability. Cloning the "perfect" animal also could capture optimum dominance and epistatic genetic effects that are otherwise difficult to select for. For some traits, maternal effects are important. In that case, clones as breeding animals must be "perfect" for both direct and maternal genotypes, or alternatively terminal and maternal clone lines would need to be developed. The use of clones to increase uniformity can be only partially successful. If heritability is 25%, then the standard deviation among clones would be 87% of that of uncloned animals. Only if heritability is 100% will clone mates have complete uniformity. Fixing the genotype could increase susceptibility to failure if environment changes or if the cloned genotype is susceptible to a new disease or if economic conditions change. Cloning, at best, is another tool for animal improvement that joins the list of previous biotechnological inventions, some of which have become cost effective, such as artificial insemination, sexing of semen, multiple ovulation and embryo transfer, embryo sexing, and in vitro fertilization. Cloning has a place in that inventory but, in the long-term, the use of cloning will need to be managed to be cost effective for the improvement of quantitative characters. PMID- 15526787 TI - Effects of supplementation on voluntary forage intake, diet digestibility, and animal performance. AB - A data base was constructed to describe and estimate supplementation effects in nonlactating cattle consuming forage ad libitum. The data base included 66 publications on 126 forages (73 harvested and 53 grazed) and a total of 444 comparisons between a control, unsupplemented treatment and a supplemented treatment. Daily gains were reported for 301 comparisons and voluntary intake for 258. Direct measures of forage digestibility were reported for 202 comparisons, and total diet digestibility for 150. Supplements did not increase gain in all cases. Change in ADG due to supplement was not related closely to intake of supplemental TDN. Lowest increases in ADG were with native forages supplemented with molasses alone or with low intakes of molasses containing high levels of NPN. Greatest increases in gain were with improved forages, supplements with > 60% TDN, and supplemental CP intake > .05% of BW. Supplements decreased voluntary forage intake (VFI) when supplemental TDN intake was > .7% of BW, forage TDN:CP ratio was < 7 (adequate N), or VFI when fed alone was > 1.75% of BW. When supplements increased VFI, forage TDN: CP ratio was > 7 (N deficit), and VFI when fed alone was often low. There was little relationship between change in VFI and sources of supplemental CP and TDN. Supplements caused total diet TDN concentration to deviate from expected values by -10 to +5% of OM. When supplemental TDN intake was > .7% of BW, diet TDN concentration was always less than expected. There was little relationship between deviation from expected total diet TDN and type or composition of forages or supplements. Empirical multiple regression equations were developed to estimate effects of supplements on VFI and total diet TDN concentration. The most acceptable intake equation estimated VFI when fed with supplement (r2 = .84) That equation included VFI when fed alone, supplement intake, CP and TDN concentrations in forage and supplement, and classification codes describing forages and supplemental energy. The most acceptable equation for estimating total diet TDN concentration included only the expected total diet TDN concentration (r2 = .87). These equations may be used in nutritional models to account for associative effects. PMID- 15526788 TI - Effects of previous grazing nutrition and management on feedlot performance of cattle. AB - Management strategies designed to improve grazing animal performance can influence feedlot performance and carcass traits both positively and negatively. In spite of the economic relevance of potential interactions between grazing and finishing performance, controlled experiments evaluating integrated production systems are limited in number. Effects of grazing treatments can result from, or be overshadowed by, changes in gut fill, thus making it difficult to assign precise costs to different phases of production. Published reports have considered the effects of stocking rate, duration of grazing, forage characteristics, supplementation, and growth-promoting implants on subsequent finishing performance. Improvements in cattle performance attributed to changes in stocking rate generally have been neutral to positive with respect to effects on finishing performance. Comparisons among forages have led to the suggestion that forage species may contribute to differences in gastrointestinal fill of grazing cattle, thereby influencing gain and efficiency during the subsequent finishing phase. Creep-feeding suckling calves generally has increased preweaning performance but has had relatively little influence on performance during the subsequent finishing phase. Grain supplementation of stocker cattle during the grazing period has improved grazing performance, but effects on subsequent feedlot performance have been inconsistent. Potential carryover effects from protein and mineral supplementation also have been inconclusive. Lack of congruence among studies is puzzling but may be the consequence of highly varied production systems, differences in experimental procedures, and changes in gut fill or mass of internal organs. Based on the studies reviewed, the expression or absence of compensatory growth during the finishing phase appears to be related to the nutritional quality of forages utilized in the grazing period, with higher quality forages tending to yield greater compensatory effects. The bulk of evidence with suckling cattle and stocker implants suggests that effects on subsequent finishing performance are minimal. Attention is drawn to the noticeable lack of research pertaining to integrated production systems. A more thorough understanding of the interactions among grazing nutrition and management, finishing performance, and carcass traits is needed to facilitate greater economic exploitation of these relationships. PMID- 15526789 TI - Applications in survival analysis. AB - Survival or failure time traits such as herd life and days open are both important economically and pose a number of challenges to an analysis based on linear mixed models. The main features of a survival trait are that it is the time until some event occurs, and some of the observations are censored. Survival models and the associated estimation procedures provide a flexible means of modeling survival traits. In this paper I will discuss the application of survival analysis based on the Weibull distribution. The components that make up a survival model will be presented along with their interpretation. Issues related to the model construction and estimation will be presented. PMID- 15526790 TI - An appropriate role for ethics in teaching contemporary issues. AB - There seems to be general agreement among animal scientists that training in contemporary issues is an appropriate goal of animal science curricula. One approach is based on the view that public policy issues embody moral dilemmas that arise when the moral perspectives (values) of opposing groups are incompatible. Based on this view, an understanding of such issues requires some training in ethics. More specifically, knowledge of major moral theories and the nature of values is necessary to identify and analyze the moral components of issues. An appreciation for ethics also provides insight into addressing issues in a morally responsible manner. The plurality of values and the contextual nature of policy issues precludes solutions based on application of universal moral principles. Nevertheless, informed judgments, based on the collective wisdom of individuals, frequently offer solutions that address opposing values. Such judgments can be attained when those affected by issues form communities, attempt to achieve a collective understanding of the problem, and, if possible, construct a common set of values that facilitates consensus. Based on this analysis, issues courses should facilitate the development of learning communities wherein students and teachers critically analyze (i.e., deconstruct) and reconstruct issues in ways that enhance understanding and instill a respect for diverse moral perspectives. An expression of such understanding and respect is the ability to distinguish between situations when consensus offers fair solutions and when it subjects minority groups to the tyranny of the majority. PMID- 15526791 TI - Team approach to teaching participatory group process involving natural resources and agriculture. AB - Contemporary issues are defined by people who share diverse and often strongly defended views about the topic. In Oregon, citizens are increasingly being asked or expected to participate in complex decisions that require a consensus. Rather than teach one professor's synthesis of a contemporary natural resource issue, faculty from six disciplines coach group process, interactive learning skills, and systems thinking as a way to address complex issues from multiple perspectives. Students learn by grappling with a natural resource issue of their choice within groups based on a diversity among majors, degree status, and gender. Students define situation (S), brainstorm new or different targets (T), and analyze two or more pathways (P), using an STP learning and action process. Exploring potential pathways involves defining possible consequences, stakeholder views, feasibility (ecological, social, economic, and political), and planning that includes expected behavior of the improved system over time. Students present their topics and improvements showing systemic relationships, systematic analysis, and integration of scientific facts and secondary data at midterm and during finals. Reflective learning is fostered throughout the course with prompted questions in a journal notebook. Grading criteria promote meaningful inquiry and participation in group process combined with integration of scientific facts and reflective learning. PMID- 15526792 TI - Odor control for livestock systems. AB - Odors are generated primarily as the result of manure storage but also result from animal housing and manure application. Effective odor control is dependent upon implementation of strategies that are complementary to management practices. Some systems use a deep pit or a holding tank for manure storage. In such systems, little or no biological processing occurs, and they are therefore considered high-load systems. In systems where biological processing occurs to a great extent, such as in anaerobic digesters or lagoons, the system would be termed a low-load system. Odor control strategies for manure storage areas, such as solids separation and additives, are best suited for low-load systems, whereas covers and biofilters provide the best results for high-load systems. Strategies that reduce nutrient production, such as dietary restriction of nutrients, are well-suited for all types of manure storage systems. To comply with current or pending odor control regulations, it is imperative that producers be provided with sound recommendations of odor control strategies. PMID- 15526793 TI - Interpretation and design of nonregulatory on-farm feeding trials. AB - Nonregulatory feeding trials are used to determine the frequency and magnitude of response to a nutritional treatment, the odds of success, and factors that influence the odds and to provide an estimate of potential economic impact. These types of trials are not appropriate for elucidating mechanisms or modes of action. Correct experimental design is critical. Important factors that impact the validity of these trials are animal randomization, potential confounding with time, identification of the proper experimental unit, and adequate replication. Animals should be assigned to treatment without a systematic influence of environmental or physiological factors. The experimental design must account for and remove these influences. The experimental unit is the smallest entity to which the treatment can be applied randomly. In commercial settings, this is often a pen of animals. In a pen feeding situation, the experimental unit is the pen, although measurements may be taken on an individual animal basis. Frequency of data collection generally does not influence the treatment effect; however, it does influence the variance associated with the observations and the power to detect differences in treatments. Statistical methods that account for repeated observations over time are required when analyzing data with multiple observations on the same animal. Improper accounting for environmental changes over time is the most common error. Other errors include improper assignment of animals to treatment, failure to replicate the treatment across multiple experimental units, lack of on-farm oversight, and poor calibration of test equipment. PMID- 15526794 TI - The regulation of growth plate cartilage turnover. AB - The advances made in the areas of genetics and nutrition during this century have resulted in improved growth rates for livestock. However, one drawback has been the increased prevalence of long bone growth problems, such as rickets, avian tibial dyschondroplasia, and osteochondrosis. Growth plate cartilage, which regulates long bone development, must maintain a tightly controlled balance between cartilage synthesis and degradation as well as chondrocyte proliferation and apoptosis. This paper will briefly review the various nutritional factors, cell signals, and proteins that help regulate growth plate chondrocytes. Some of the growth plate diseases will be discussed with an emphasis on how a breakdown in growth plate metabolism is related to the observed problems. The author's intention is that readers will gain an appreciation for the complexity of this relatively small tissue and for why a better understanding of its physiology will be important in the years to come for the prevention of skeletal problems related to long bone growth. PMID- 15526795 TI - Bone: more than a stick. AB - Despite the importance of the skeleton for animal welfare, research into the skeletal biology of farmed species is relatively neglected. This review describes basic concepts relating to current knowledge of the control processes responsible for bone growth and its subsequent remodeling from a physiological, cell biological, and mechanical function perspective. PMID- 15526796 TI - Nutritional benefits of specialty corn grain hybrids in dairy diets. AB - Corn grain is a primary energy supplement in dairy diets and can contribute up to 30, 60, and 98% of the diet's protein, net energy, and starch, respectively. Specialty corn hybrids are one result of efforts to select corn based on nutrient content. Many of these hybrids, which include high lysine, high oil, waxy, white, and sugary, among others, have been the subject of renewed interest because of improvements in agronomic performance, commitments by marketers to preserve the identity of specialty grain, and improvements in our understanding of digestion and nutrient requirements. Nutrients targeted in corn for dairy cattle include protein content, amino acid quality (especially lysine and methionine that escape ruminal fermentation), starch (including form, texture, and digestibility), lipid content, fatty acid composition, and mineral composition and availability. Concentrations of protein and oil are highest in the germ, and both are negatively correlated with starch concentration. Efforts to improve lysine content are hampered by its negative correlation with total protein. Lactation studies to date have shown variable production responses to specialty corns; their value may be limited to replacing more expensive ingredients in dairy diets. Increasing the protein content of corn would have the most economic value in diets for cows producing less than 9500 kg per lactation. High lysine corn is currently of limited value; increases in ruminally undegraded lysine would increase its value in diets for cows producing more than 9500 kg per lactation. At current fat ingredient prices, high oil corn has limited economic value except in diets for very high production. PMID- 15526797 TI - Nutritional benefits of specialty grain hybrids in beef feedlot diets. AB - The study of grain hybrids with faster or more extensive rates of ruminal starch fermentation has been a key research area. Because grain sorghum starch is generally regarded as less accessible to enzymatic degradation than starch in other grains, it has received the greatest research emphasis. However, all grains have been evaluated to some extent. Grain sorghum hybrids appear to be more variable in digestibility, in vitro and in vivo, and in rate of starch fermentation than are corn hybrids. The greater variation may be partially because grain sorghum hybrids are developed and evaluated under more stressful environmental conditions (high temperature and limited water conditions) than are corn hybrids. In vitro and in vivo studies indicate that differences in grain hybrids exist, but these differences may not totally explain differences in cattle performance. The response to feeding high-lysine corn to cattle has been variable. Although high-lysine corn supplies more lysine in the diet, lysine flow to the abomasum was not increased. Hybrids selected for increased lysine content have been shown to have faster in vitro rates of starch digestion, suggesting that improvements in animal performance may be related to the indirect selection for improved energy utilization. In one study in which high-oil corn was evaluated, feed conversion was not improved compared with a control corn diet. At the present time additional studies in which other genetic modifications of grain hybrids are evaluated are in progress, but the results have not been published. PMID- 15526798 TI - The Beltsville sperm sexing technology: high-speed sperm sorting gives improved sperm output for in vitro fertilization and AI. AB - The Beltsville sperm sexing technology is currently the only effective means of altering the sex ratio of offspring in livestock. The method is based on the flow cytometric separation of X- and Y-chromosome-bearing sperm based on X/Y DNA content difference. It is an effective means of producing progeny of predetermined sex in cattle, swine, sheep, and laboratory animals. The method involves treating sperm with a DNA-binding fluorochrome, Hoechst 33342, and flow cytometrically sorting them into separate X and Y populations that can subsequently be used for surgical intratubal or intrauterine insemination, deep uterine insemination, regular artificial insemination in some cases, in vitro fertilization to produce sexed embryos for transfer, and intracytoplasmic sperm injection of ova. Skewed sex ratios of 85 to 95% of one sex or the other have been repeatably achieved in most species. The method has been used worldwide to produce several hundred morphologically normal animal offspring of the predicted sex. It has also been validated in the laboratory using DNA reanalysis of the sorted sperm populations and by fluorescence in situ hybridization and PCR of individual sperm. We developed a new orienting nozzle that we have fitted to both conventional and high-speed cell sorters that have been modified for sperm sorting. Recently we completed the adaptation of the new orienting nozzle to a Cytomation MoFlo high-speed cell sorter modified for sperm. This adaptation of the nozzle has increased the overall production rate of sorted X and Y sperm from about .35 million/h to 5 or 6 million sperm/h (each population). Calves have been born from cows artificially inseminated using conventional technique and sexed sperm. In addition, numerous litters of pigs have been born after transfer of embryos produced from X or Y sorted sperm. PMID- 15526799 TI - The callipyge phenomenon: evidence for unusual genetic inheritance. AB - In 1983, a male lamb exhibiting a pronounced muscular hypertrophy, particularly noticeable in the hind quarters, was born into a commercial Dorset flock in Oklahoma. The ram was premonitorily called Solid Gold. He subsequently produced offspring expressing the unusual phenotype, which is referred to as callipyge (Greek: calli- beautiful + -pyge buttocks). Animals demonstrating the callipyge phenotype are all descendants of this founder ram. These animals produce leaner, higher yielding carcasses, but there is some concern with decreased tenderness of the loin. Genetic characterization of the locus has demonstrated a unique mode of inheritance termed polar overdominance, in which only heterozygous offspring inheriting the mutation from their sire express the phenotype. The three other genotypes are normal in appearance. Progeny data indicate that reactivation of the maternal callipyge allele occurs after passage through the male germ line, although this reactivation is not absolute. The callipyge gene has been mapped to the distal end of ovine chromosome 18. PMID- 15526800 TI - Basics of gametic imprinting. AB - The fundamental assumption of Mendelian genetics is that behavior of an allele is identical whether it arrives to a zygote through paternal or maternal germline pathway. Gametic imprinting phenomena discovered and studied in mammals show limitations of the classical view in special cases. Two sources of evidence were essential to describe gametic imprinting. The first approach based on genetic evidence demonstrated that some maternally and paternally derived regions of certain chromosomes were not equivalent. Paternal or maternal disomy of the regions containing particular genes caused significant effects on viability and development of progeny. The second set of data was obtained by nuclear transplantations and parthenogenetic activation of mammalian oocytes. These data suggested that the contribution of parental genomes was not equivalent and differential imprinting of nuclear genes during gametogenesis was very likely. The number of loci found in mice, which show gametic imprinting, is 34 and continues to grow. It is generally accepted that gametic imprinting is a mammalian invention and there are differences in imprinting pattern between species. Most hypotheses propose involvement of imprinted genes in the control of fetal growth and fetal-maternal interactions, thus keeping a balance between contradictory fetal and maternal requirements. Molecular mechanisms responsible for gametic imprinting still remain to be studied, but for several genes it was shown that imprinting marks are imposed by a parent-specific methylation process during gametogenesis. These marks are resistant to global demethylation during cleavage and to global de novo methylation after implantation and maintain different methylation patterns in paternal and maternal alleles of imprinted genes. About 20 to 25% of all transgene loci studied demonstrate similarities with imprinted genes. For instance, methylation of some transgenes is dependent on parental gametic pathway and reversible in the next generation. There are data indicating that selection of modifier genes may change the effects of gametic imprinting. It is possible that future selection and crossbreeding programs may take gametic imprinting into consideration. PMID- 15526801 TI - The callipyge phenomenon: tenderness intervention methods. AB - Consumers continue to desire leaner meats. Lambs expressing the callipyge gene have been identified as having superior, leaner carcasses compared with normally muscled lambs. However, the longissimus muscle, a major merchandised muscle in lamb, has repeatedly been shown to be significantly less tender in callipyge lamb compared with normally muscled lambs. Preharvest factors, such as genetics, sex, and production/management practices, have thus far shown no promise at alleviating this tenderness problem. But a number of postharvest interventions have been introduced to alleviate it. Included among the strategies are postmortem aging, carcass electrical stimulation, the combination of freezing and thawing before aging, calcium chloride injection, and the Hydrodyne process. These strategies have exhibited various degrees of success. Postharvest strategies to improve callipyge longissimus tenderness are described in this article. PMID- 15526802 TI - Economics of callipyge lamb production. AB - This paper examines the economic implications of callipyge (CLPG) lamb production. The price, as it relates to competing meats and excess fat, significantly impact lamb demand, and CLPG genetics improves those factors. The CLPG phenotype does not affect number or weight of lambs weaned or postweaning ADG, but it does improve postweaning feed efficiency by approximately 10%; dressing percentage approximately 7.5%; and yields of wholesale leg (11.8%), loin (4.7%), rack (2.5%), and shoulder (2.3%). Total production costs for a 59-kg lamb are 4% lower in CLPG lambs due to improved feed efficiency. Assuming pelt and offal value pays for slaughter costs, the costs of normal (N) and CLPG carcasses are the same as for live lambs, $81 and $78, respectively; but, due to dressing percentage, the N carcass weighs 29.2 kg and the CLPG carcass, 31.4 kg. Thus, carcass costs for N and CLPG lambs are $2.77/kg and $2.49/kg, respectively. Decreased feed costs, combined with increased carcass and primal cut yields for CLPG lambs, lowers the price required to recover meat costs for leg, loin, rack, and shoulder by 19.7, 14.4, 12.6, and 11.9%, respectively. Successful marketing of CLPG loin and rack depends on the use of one of several postharvest tenderization procedures. Moisture-enhanced pork is accepted by consumers and often sells for a premium; and moisture enhancement may be appropriate for CLPG lamb. The meat cost per kilogram (including a $.10 per kilogram treatment cost) of tenderized and moisture-enhanced CLPG leg, loin, rack, and shoulder containing 10% added water and ingredients would be lowered to $2.51, $4.65, $5.34, and $1.85, respectively. That represents a total of a 20.9% reduction in cost-basis price. When expressed on the basis of increased revenue from the additional yield of cuts at a given market price, the value of CLPG and moisture-enhanced CLPG cuts from a 59-kg lamb would be, respectively, 14.2% and 23.4% higher than for N lamb. Industrywide adoption of CLPG could increase intermediate-run U.S. profits by $109 million, but the actual effects of CLPG attributes, such as a visual appeal, lower fat and cholesterol content, and reduced seam fat, on consumer demand need to be quantified. If accepted by packers and consumers, moisture enhanced CLPG lamb has the potential to decrease the cost of lamb to consumers and increase lamb industry profitability. PMID- 15526803 TI - Corticosteroid and immunomodulatory agents in idiopathic pulmonary fibrosis. AB - Idiopathic pulmonary fibrosis (IPF) is a progressive pulmonary disease leading to death within a few years of diagnosis despite medical therapy. On the basis of methodologies of the Cochrane collaboration, this overview discusses the evidence for IPF therapy. Good-quality studies on oral corticosteroids, the most common medical therapy in use for IPF, are lacking. A few small studies have been carried out on the efficacy of many non-steroid immunosuppressive agents, and the results have been generally disappointing. The most extensively studied medical therapy, gamma interferon, showed a significant effect in a small randomized study, but its efficacy was not confirmed in a larger randomized-controlled trial. The long-awaited good news for patients affected by this deadly disease, and for their physicians, could come in the near future from large randomized controlled trials with gamma interferon or other immunomodulatory agents. PMID- 15526804 TI - Gender does not influence the response to the combination of salmeterol and fluticasone propionate in COPD. AB - The prevalence of chronic obstructive pulmonary disease (COPD) in women is increasing worldwide. Women may have greater susceptibility to COPD progression than men, and differences in efficacy and safety of respiratory medications by gender are largely unexplored. We aimed to determine whether the response to treatment in women with COPD differed from men in a large, 1-year double-blind trial ('TRISTAN'). In a sensitivity analysis, we compared 539 male and 180 female COPD patients, who were randomized to the saLmeterol/fluticasone combination 50/500mcg bid or placebo for 12 months. Combination therapy improved pre treatment FEV1 significantly more than placebo in women by 152 ml (95% confidence interval 95-208) and in men by 127 ml (94-159). Similarly, a reduction in COPD exacerbation rates of 31% in women (9-48%) and of 23% in men (8-35%) was observed. Combination therapy reduced COPD exacerbations requiring treatment with oral corticosteroids by 36% in women and by 41% in men. Finally, combination treatment produced a better improvement in health status than placebo with a decrease in the SGRQ scores in women by -2.3 (-4.6 - 0.1) and in men by -2.1 ( 3.5 to -0.8). No gender interaction was found for any outcome. Treatments were well tolerated with no difference in the frequency of adverse events in women and men. In this trial, therapy with the salmeterol/fluticasone combination produced significant improvements compared to placebo on all main endpoints and the magnitude of these improvements was similar for both men and women. PMID- 15526805 TI - A comparison of the effects of oral montelukast and inhaled salmeterol on response to rescue bronchodilation after challenge. AB - OBJECTIVES: To compare the effects of addition of montelukast or salmeterol to inhaled corticosteroids (ICS) on the response to rescue beta2-agonist use after exercise-induced bronchoconstriction. METHODS: A double-blind, placebo-controlled study was performed at 16 centers in the United States. Patients with asthma (n = 122, ages 15-58) whose symptoms were uncontrolled on Low-dose inhaled fluticasone and who had a history of exercise-induced worsening of asthma were randomized to receive either montelukast (10 mg once daily), salmeterol (50microg twice daily), or placebo for 4 weeks. Standardized spirometry after exercise challenge and beta2-agonist rescue was performed at baseline, week 1 and 4. RESULTS: Maximum achievable forced expiratory volume in 1 s (FEV1) percent predicted after rescue beta2-agonist improved in the montelukast (+1.5%) and placebo (+1.2%) groups at 4 weeks, but diminished in the salmeterol (-3.9%) group (P < 0.001). Although pre exercise FEV1 was greatest with salmeterol (P = 0.10), patients taking montelukast had significantly greater protection from an exercise-induced decrease in FEV1 than those taking salmeterol (P < 0.001). Both the magnitude and rate of rescue bronchodilation were greater with montelukast compared with salmeterol (P < 0.001). Five minutes after rescue beta2-agonist, 92% of patients taking montelukast and 68% of those taking placebo had recovered to pre-exercise levels, whereas only 50% of those taking salmeterol had recovered to pre-exercise levels. CONCLUSION: In patients whose asthma symptoms remain uncontrolled using ICS, addition of montelukast permits a greater and more rapid rescue bronchodilation with a short-acting beta2-agonist than addition of salmeterol and provides consistent and clinically meaningful protection against exercise-induced bronchoconstriction. PMID- 15526806 TI - Effect of respiratory rate on airway deadspace ventilation during exercise in cystic fibrosis. AB - Gas exchange during exercise in patients with cystic fibrosis (CF) is characterised by an elevated physiological deadspace to tidal volume ratio. While this has been attributed to alveolar ventilation perfusion mismatch, there are other potential causes of the high proportion of wasted ventilation, including factors relating to the volume and the ventilation of the airway deadspace. CF (n = 6, F = 1, FEV1 26-63% pred) and control (n = 6, F = 2) subjects completed steady-state exercise on a cycle ergometer. Gas exchange was measured breath-by breath and the volume of the airway deadspace (V(Daw)) determined using the equal areas method. Exercise data were interpolated to a CO2 output of 0.7 l/min. V(Daw) was similar in the two groups both at rest and during exercise. However, the airway deadspace ventilation (V(Daw)) (median (inter-quartile range)), patients, 6.8 (5.1-7.1) l/min; controls, 4.9 (3.5-5.6) l/min, P < 0.05) was significantly greater in the CF group due to a greater respiratory frequency. These results indicate that in CF patients, abnormally increased V(Daw) is an important contributor to the total (physiological) deadspace ventilation. Exercise performance in CF might be enhanced by efforts directed at facilitating an increase in exercise tidal volume and therefore the adoption of a more efficient pattern of breathing. PMID- 15526807 TI - Factors determining performance of bronchodilator reversibility tests in middle aged and elderly. AB - Knowledge of the factors determining the performance of bronchodilator reversibility testing in a general population is lacking. Random samples of all adults aged 47-48 and 71-73 years living in Bergen, Norway, were invited to a cross-sectional study. Altogether 3506 subjects attended (69%). Test quality predictors were examined through multiple logistic regression analyses including gender, age, body mass, educational level, smoking history, respiratory symptoms, and in the elderly cohort cognitive level. Among the participants, 1.6% refused to inhale salbutamol, 2.5% failed the initial spirometry according to the ATS guidelines, and 1.3% failed the post-bronchodilator spirometry. Old age and body mass index > 30 kg/m2 were independent risk factors for an unsuccessful initial spirometry, and never smoking and respiratory symptoms were risk factors for failing the post-bronchodilator spirometry. Cognitive impairment in the elderly was a risk factor for failing both the initial- and post-bronchodilator spirometry. The median number of forced expirations was 7 in subjects obtaining an acceptable reversibility test. One third of these participants needed > or = 8 attempts, with independent predicting factors being old age, Little formal education and never smoking. Although reversibility testing becomes increasingly difficult with age, reliable data are obtained in a vast majority (94%) of subjects in our community study. PMID- 15526808 TI - Chemo-radiotherapy for stage III unresectable non-small cell lung cancer long term results of a prospective study. AB - INTRODUCTION: Combined-modality treatment is considered standard of care in the treatment of stage III non-small-cell lung cancer (NSCLC). This study was designed to assess the efficacy and tolerability of induction paclitaxel/carboplatin followed by concurrent thoracic radiotherapy and weekly paclitaxel. MATERIALS AND METHODS: Patients with unresectable stage III NSCLC were treated prospectively with two cycles of paclitaxel (175 mg/m2) and carboplatin (area under the curve of 6) followed by radiotherapy (60-66 Gy) concurrent with 6 weekly doses of paclitaxel (60 mg/m2). Response was determined 8 weeks after the completion of treatment and treatment-related toxicities were assessed at each visit during treatment and follow-up. RESULTS: Sixty-three patients were treated, 5 had complete response and 33 had partial response, giving a response rate of 60%. Thirty-seven percent of patients developed grade 3 or 4 neutropenia; 48% had significant esophagitis requiring the use of narcotic analgesics. Two patients developed esophageal stricture subsequently. The median survival was 51 months and 12 months for stage IIIA and IIIB patients, respectively. Progression-free survival was 16 months and 11 months respectively. CONCLUSIONS: The response rate was encouraging. Esophagitis was a significant morbidity and should prompt modification of treatment regimen, either in the chemotherapy schedule or by adjusting the radiotherapy treatment planning. PMID- 15526809 TI - LPS-induced bronchoalveolar neutrophilia; effects of salmeterol treatment. AB - Salmeterol has earlier been reported to have immune modulating effects on Lipopolysaccharide (LPS)-induced neutrophilic inflammation in rodents. The aim of this study was to explore whether 3 weeks regular treatment with inhaled Salmeterol would have a protective effect against neutrophilia, following an LPS inhalation as assessed by bronchoscopy with bronchial wash (BW) and bronchoalveolar lavage (BAL) in healthy subjects. Fifteen volunteers all underwent bronchoscopies with bronchial wash and BAL on three occasions, each being 3 h after inhalation provocation. The initial inhalation was with saline (dilutant) as a reference and the two following with LPS 50 microg diluted in saline. After the saline inhalation the subjects were randomised to treatment with Salmeterol 50 microg twice daily and placebo in a double-blind double-dummy crossover design. Compared to saline inhalation, the LPS inhalations resulted in a two-fold increase in neutrophils both in BW and BAL, respectively (P < or = 0.01). The neutrophilia was present irrespective of the LPS inhalation was preceded by placebo or Salmeterol. This experimental study could not confirm any modulating effect of Salmeterol on LPS-induced airway neutrophilia. PMID- 15526810 TI - Antibiotic prescribing rates in the US ambulatory care setting for patients diagnosed with influenza, 1997-2001. AB - To document the rate and cost of antibiotic prescribing for patients diagnosed only with influenza during US ambulatory care visits. Federal survey data for 1997-2001 were used to estimate outpatient trends for all patients and healthy people age 5-49 years. Cost estimates were based on Medicare payments and Red Book average wholesale prices in 2003. Antibiotic prescribing for influenza is widespread; 38% of visits led to an antibiotic prescription of which one-third were for broad spectrum antibiotics. Inappropriate antibiotics cost dollar 18.5 million annually and may contribute to resistance. Increased vaccination rates and viral testing could reduce these trends. PMID- 15526811 TI - Pre-medication for bronchoscopy: a randomised double blind trial comparing alfentanil with midazolam. AB - This study was designed to compare the effects of alfentanil and midazolam pre medication on patient comfort during and after flexible bronchoscopy. A randomised, double-blind study was performed; 40 patients received alfentanil and 29 midazolam. Subjects completed questionnaires about discomfort and adverse effects immediately post-procedure and 24 h later. The bronchoscopist also completed a questionnaire. No difference in patient discomfort was found immediately post-procedure and no differences were found for amount of topical lignocaine used or minimum oxygen saturation. Operators reported no overall difference between the agents for ease of procedure but about 20% less cough was reported in the alfentanil group (P = 0.02). Patient discomfort scores in the 24h questionnaire were significantly lower in patients given midazolam (P = 0.01 for nasal discomfort, P = 0.003 for throat discomfort) but drowsiness was commoner in this group (P = 0.04). There was no significant difference in patients' reports of cough, nausea or vomiting or their willingness to have a repeat procedure. In conclusion, cough during bronchoscopy was slightly less marked with alfentanil than midazolam pre-medication but this made no difference to the ease of procedure or to overall patient discomfort. Patients given midazolam reported less discomfort when asked about the test 24 h later. PMID- 15526812 TI - Prevalence and incidence of asthma related to waist circumference and BMI in a Swedish community sample. AB - Both asthma and obesity have become more common in affluent societies during the recent decades and several studies have shown a correlation between the presence of asthma and obesity. In order to further study this association we have investigated a population from a community in southern Sweden, where almost all inhabitants had their body indices measured as part of a study on diabetes at a primary care centre. An asthma unit working with a structured care programme for asthma was available. This organisation enabled us to study whether body mass index and waist circumference was associated with having or developing asthma. There was a significant association between both overweight, increased waist circumference and asthma, P < 0.01. The risk for developing asthma was associated with increased body weight and abdominal circumference, P < 0.05. The increase in asthma morbidity in the overweight subjects was found almost exclusively in the non-atopic asthma patients. This study confirms earlier findings of an increased prevalence of asthma in obese and overweight patients. Increased obesity and especially abdominal obesity is thus a risk factor for asthma, which probably contributes to the high prevalence of asthma in affluent societies. PMID- 15526813 TI - Red cell macrocytosis in hypoxemic patients with chronic obstructive pulmonary disease. AB - Macrocytosis is a common finding in patients with chronic obstructive pulmonary disease (COPD). The cause for the elevation of mean corpuscular volume (MCV) in these patients remains elusive. In an attempt to determine the extent of macrocytosis in COPD patients and search for possible causative factors, we evaluated the hematologic parameters, F-cell percentage, blood gases and serum erythropoietin (Epo) Levels in 32 COPD clinically stable patients and 34 sex- and age-matched non-smoker healthy volunteers. An increased MCV was observed in almost half of the hypoxemic COPD cases (14/32 or 43.75%), while erythrocytosis developed to a lesser degree (37.5%). The erythropoietic response did not correlate with the severity of hypoxia. Moreover, no significant correlation was found between macrocytosis and hypoxemia or erythrocytosis and red cell size. In some cases the two phenomena occurred independently. The F-cell percentage was significantly elevated in the COPD group (P < 0.01) and was associated with MCV values (n = 32, r5 = 0.41, P < 0.05). This finding supports the hypothesis we put forward to explain the macrocytosis often observed in COPD, i.e., that the acute erythropoietic stress occurring repeatedly in these patients as a result of the frequent exacerbations may lead to waves of release of relatively immature, large red cells from the marrow, including an increased number of F-cells, reflecting the recruitment of normally dormant BFU-E (bursts forming units of erythrocyte precursors), which maintain the program for gamma-chain synthesis. The fact that erythrocytosis and macrocytosis, both being triggered by hypoxemia, do not occur consistently in all COPD patients indicates that many other factors may also intervene. PMID- 15526814 TI - Impact of spirometry on GPs' diagnostic differentiation and decision-making. AB - BACKGROUND: Spirometry is increasingly implemented in general practice, while the ability of general practitioners (GPs) to interpret flow volume curves (F-V curves) has been questioned. Furthermore, the role of spirometry in the GPs decision-making process has barely been studied. AIM: To compare the achievements of trained GPs in spirometric diagnosis with an expert consensus panel (1) and to assess the influence of spirometry on the GPs decision-making (2). METHOD: Twelve cases including a wide range of F-V curves were interpreted by 39 GPs as well as the expert panel. Diagnostic test characteristics were calculated using multi level analysis and summarised by diagnostic odds ratios (DOR). Differences in decision-making indicators were expressed as odds ratios and 95% confidence intervals. RESULTS: Normal F-V curves (DOR 65.0) and obstructive F-V curves (DOR 48.9) were reasonably well diagnosed, while rare and mixed pathological patterns achieved considerably lower scores (DOR 3.8). Intermediate scores were obtained in the recognition of incorrect test manoeuvres (DOR 24.4). Spirometry influenced the GPs decision-making in reducing the number of alternative diagnoses (OR 0.266 [0.200, 0.353]), but also increased referral rates (7.26 [4.71, 11.2]) and the use of diagnostic prednisolone courses (4.55 [3.12, 6.64]) substantially. CONCLUSION: Trained GPs were able to differentiate between normal and obstructive disease patterns, while F-V curves suggestive of rare and mixed pathology were often missed. Spirometry seems to influence the decision-making process of the GP; whether this represents an initial or a more sustained effect remains to be evaluated in studies of daily primary care practice. PMID- 15526815 TI - Role of KL-6 in evaluating the disease severity of rheumatoid lung disease: comparison with HRCT. AB - OBJECTIVE: To determine the role of KL-6 (Krebs von den Lungen-6) in evaluating the disease severity of pulmonary lesions in rheumatoid arthritis (RA) compared with high resolution computed tomography (HRCT) findings. METHODS: Fifty serum KL 6 levels and HRCT images were prospectively obtained from 47 RA patients. Eight HRCT findings were classified into five grades. Patients were also divided into two groups according to the KL-6 threshold level and HRCT findings were evaluated. RESULTS: There was a positive correlation between the serum KL-6 level and the total CT score (r = 0.83). Reticular opacity most closely related to the serum KL-6 levels (r = 0.84). In the high KL-6 group (n = 10), the average CTscore was markedly increased (64.6 points) and severe honeycombing expanded into the whole lung. One case revealed diffuse ground glass opacity. In 12 of 40 cases in the normal KL-6 group, CT scores mildly increased compared with the other cases (over 20 points). The predominant finding of these cases could be classified into four types: (1) narrow spread honeycombing; (2) subtle fibrosis; (3) airway diseases; and (4) dense consolidation. CONCLUSION: KL-6 is a useful marker to detect severe RA lung disease. It is also useful to distinguish non fibrosis from fibrosis predominant cases. However, it sometimes could not detect early stage RA lung disease. PMID- 15526816 TI - The response to beta-agonists in wheezy infants: three methods compared. AB - BACKGROUND: Studies into the effects of salbutamol in the treatment of wheeze in infancy have been conflicting, possibly due to differences in outcome variables. We aimed to assess the response to salbutamol using indices derived from passive and forced expiration. METHODS: We recruited 39 infants who had a history of wheezing (mean age 43 weeks) and measured maximum flow at functional residual capacity (V'(max FRC)) by rapid thoracoabdominal compression (RTC), and forced expired volume at 0.4s (FEV0.4) using the raised-volume RTC technique (RV-RTC). We calculated passive compliance (C(rs)), resistance (R(rs)) and time constant (tau) from relaxed expirations that followed the augmented inspirations delivered during RV-RTC. Measurements were repeated after aerosol salbutamol (800 mcg). RESULTS: Data were obtained in 32 infants for V'(max FRC), 22 for FEV0.4 and 19 for passive mechanics. There were no mean changes in any index of forced expiration after salbutamol. Some individuals showed significant changes (improvement or worsening) in one or other index. Overall, there was a small increase in C(rs) after salbutamol but no change in R(rs) or tau. CONCLUSIONS: We found no consistent pattern of response in either index of forced expiration. Validated clinical scores or alternative physiological techniques may be preferable to respiratory mechanics in assessing bronchodilator response. PMID- 15526817 TI - An economic evaluation of combination treatment with budesonide and formoterol in patients with mild-to-moderate persistent asthma. AB - Patients with mild asthma may benefit from increasing their inhaled corticosteroid dose, adding a long-acting beta2-agonist, or both. This study assessed the cost-effectiveness of these options. Patients aged > or = 12 years with mild-to-moderate persistent asthma (n = 1272) were randomised to twice daily, double-blind treatment with budesonide 100 microg, budesonide 100 microg plus formoterol 4.5 microg, budesonide 200 microg, or budesonide 200 microg plus formoterol 4.5 microg for 12 months. Clinical variables included lung function, number of symptom-free days and number of severe exacerbations. Data on medication use, hospitalisation, visits to health professionals and time off work due to asthma were combined with Swedish unit cost data (1999) to estimate the mean annual cost per patient. Budesonide 200 microg plus formoterol 4.5 microg had the greatest efficacy and effectiveness. Budesonide 200 microg plus formoterol 4.5 microg was both more effective and less costly than budesonide 100 microg plus formoterol 4.5 microg, so a cost-effectiveness ratio was not calculated for this comparison. The cost-effectiveness ratio for budesonide 200 microg plus formoterol 4.5 microg compared with budesonide 200 microg alone was SEK 21 per symptom-free days gained. The combination of budesonide and formoterol in mild-to-moderate persistent asthma improved effectiveness at modest additional cost. PMID- 15526818 TI - [Quality of life scales, the psychiatrist's point of view]. PMID- 15526819 TI - [Principal determinants of quality of life]. PMID- 15526820 TI - [The role of erythropoietin]. PMID- 15526821 TI - [How to better the quality of life of dialysed patients]. PMID- 15526823 TI - [Roche plays with dialysis professionals!]. PMID- 15526822 TI - [Pre-transplantation, what words to say]. PMID- 15526824 TI - [Infections related to central venous catheters]. PMID- 15526825 TI - [Peritoneal dialysis in 2004]. PMID- 15526826 TI - [Renal transplantation in the elderly patient]. PMID- 15526827 TI - [Psychological management of the chronic renal insufficiency patient]. PMID- 15526828 TI - Salvage therapy options for myeloma patients. PMID- 15526829 TI - Prostate cancer 2004: insights from national disease registries. AB - In 2004, the large majority of prostate cancers are detected via prostate specific antigen (PSA) screening. Most are diagnosed at an early stage and are amenable to aggressive local treatment. However, the natural history of the disease may be prolonged, and all available active treatments exert a potential negative effect on patients' HRQOL. Management options for localized prostate cancer have become increasingly complex in recent years, and rigorous trials are frequently difficult to perform due to the extended follow-up required to reach meaningful outcomes. In this context, the advent of the national prostate cancer disease registries-Prostate Cancer Outcomes Study (PCOS), Center for Prostate Disease Research (CPDR), Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE), and Shared Equal Access Regional Cancer Hospital (SEARCH)-has greatly facilitated clinical research in prostate cancer. This review summarizes key findings from the registries in the areas of risk migration, practice patterns, outcome prediction, and quality-of-life outcomes. The availability of these large databases of patients will be a tremendous asset as prostate cancer management continues to evolve in the coming years. PMID- 15526830 TI - Interdisciplinary breast cancer care: declaring and improving the standard. AB - The contemporary management of breast cancer is a complex endeavor that requires a truly collaborative team approach, characterized by ongoing communication and active information-sharing among the multiple disciplines involved. Programs designed to provide comprehensive breast cancer management by a team of multidisciplinary specialists were introduced in the late 1970s and have been increasing slowly. Patients attending comprehensive breast centers receive care from a broad-based multidisciplinary team that most often includes surgeons, radiologists, pathologists, medical oncologists, radiation oncologists, plastic/reconstructive surgeons, primary care physicians, gynecologists, nurses, social workers, patient advocates, and genetic risk counselors. At the heart of comprehensive, interdisciplinary breast care is the consensus planning conference that brings together team members on a regular basis to discuss individual patient cases and develop comprehensive treatment plans. This interactive and dynamic forum has become integral to the interdisciplinary management of breast diseases and results in an increased level of communication between the participating health-care professionals and the patients they treat. Several professional organizations, most prominently the American Society of Breast Disease, promote and support an interdisciplinary approach to breast care. PMID- 15526831 TI - Clinical trials referral resource. Current clinical trials in mesothelioma. PMID- 15526832 TI - Comparing radical prostatectomy and brachytherapy for localized prostate cancer. AB - Radical prostatectomy and ultrasound-guided transperineal brachytherapy are both commonly used for the treatment of localized prostate cancer. No randomized trials are available to compare these modalities. Therefore, the physician must rely on institutional reports of results to determine which therapy is most effective. While some investigators have concluded that both therapies are effective, others have concluded that radical prostatectomy should remain the gold standard for the treatment of this disease. This article reviews the major series available for both treatments and discusses the major controversies involved in making these comparisons. The data indicate that for low-risk disease, both treatments are effective, controlling disease in over 80% of the cases, with no evidence to support the use of one treatment over the other. Similarly, for intermediate-risk disease, the conclusion that one treatment is superior to the other cannot be drawn. Brachytherapy should be performed in conjunction with external-beam radiation therapy in this group of patients. For patients with high-risk disease, neither treatment consistently achieves biochemical control rates above 50%. Although radical prostatectomy and/or brachytherapy may play a role in the care of high-risk patients in the future, external-beam radiation therapy in combination with androgen deprivation has the best track record to date. PMID- 15526833 TI - Oxaliplatin shows significant benefit over standard therapy in early-stage colon cancer. PMID- 15526834 TI - Tositumomab/I-131 tositumomab following chemotherapy effective as first-line treatment of NHL. PMID- 15526835 TI - Amifostine reduces radiation therapy-induced dry mouth in head and neck cancer patients. PMID- 15526836 TI - Rituximab maintenance stretches progression-free survival in patients with indolent NHL. PMID- 15526837 TI - Treatment of advanced non-small-cell lung cancer in special populations. AB - The benefits of chemotherapy in non-small-cell lung cancer (NSCLC) patients remains, to some extent, restricted to younger patients with a good performance status (PS). It has long been assumed that chemotherapy is too toxic and of marginal benefit for elderly NSCLC patients and those with a PS of 2. Nevertheless, retrospective analyses and more recent prospective trials have suggested that such patients enjoy longer survival and a better quality of life when treated with chemotherapy. This article will review the data and discuss their clinical implications. PMID- 15526838 TI - [Effect of erythropoietin on tissue factor and tissue factor pathway inhibitor in uremic hemodialysis patients]. AB - Some parameters of extrinsic coagulation pathway, including concentration and activity of tissue factor and concentration of tissue factor pathway inhibitor, have been estimated in uremic hemodialysis patients. The impact of erythropoietin treatment on the extrinsic coagulation pathway has also been the aim of the study. Increased concentration of tissue factor pathway inhibitor--TFPI has been found both in dialysed and non-dialysed uremic patients. This finding may be the evidence of endothelial damage as well as the protective factor against thrombotic complications. Erythropoietin treatment seemed not to induce statistically significant changes in extrinsic coagulation pathway. Some results indicate that estimation of "truncated' and "full length" forms of TFPI may be more useful comparing to complete TFPI concentration. PMID- 15526840 TI - [Relationship between peritoneal equilibration test and surface tension of peritoneal effluent in CAPD patients]. AB - Surface phenomena resulting from interactions between molecules occur commonly in nature. Peritoneal effluent is a mixture of organic and inorganic substances both macro- and micromolecular. Surfactants present in dialysate affect its surface properties. Among them are: proteins, phospholipids, fatty acids. Our aim in this study was to investigate relationships between peritoneal membrane solute transport characteristics and surface tension of peritoneal effluent. The study was conducted in 40 CAPD patients who were stable, without peritonitis (24 M, 16F), age 51.5 +/- 15.8 (range 30-79) mean CAPD duration 26.4 +/- 20.6 months (range 4-72). Standard peritoneal equilibration test (sPET) was done in all patients. Dialysate surface tension (ST) values after 4 hours dewell were determined using Wilhelmy Plate method. Mean ST values of individual dialysate sample based on 10 measurements were calculated. According to the PET values patients were divided into two groups: group 1 (high/high average transporters, n = 26) and group 2 (low average/low transporters, n = 14). Patients in group 1 had significantly lower ST of dialysate than patients in group 2 (51.2 +/- 4.8 vs 57.9 +/- 1.4 mN/m), p<0.01. The lowest values of ST (48.5 +/- 5 mN/m) were found in patients classified as high transporters (n = 8). Correlation's: significant negative correlation was found between ST and D4/P4 for creatinine (r = -0.45, p<0.005) and significant positive correlation between ST and D4/DO for glucose (r = 0.48, p = 0.003). We conclude that there are significant relationships between peritoneal transport status and surface tension of peritoneal effluent. High transporters have significantly higher concentrations of surfactants in dialysis effluent. PMID- 15526839 TI - [Effect of salmon calcitonin on bone mineral density and calcium-phosphate metabolism in chronic hemodialysis patients with secondary hyperparathyroidism]. AB - The aim of the study was to evaluate the effect of salmon calcitonin on bone mineral density, parathyroid and thyroid C cells, and calcium-phosphate metabolism in chronic hemodialysis patients with uremic hyperparathyroidism. Forty five patients with serum 1-84 PTH >220 pg/ml were divided into 2 groups: group I (n = 25), treated with intranasal salmon calcitonin (200 IU, thrice a week) and control group II (n = 20). Patients received calcium carbonate (up to 6 g/d) alone or with aluminum hydroxide (up to 3 g/d) as phosphate binders; dialysate calcium was 1.75-2 mmol/l. The observation period was 12 months. The following parameters were measured: bone mineral density (BMD) with dual-energy X ray absorptiometry in: lumbar spine (L2-L4), femoral neck and total body, before and after the study; serum endogenous calcitonin, before and after the study; serum PTH, alkaline phosphatase and total hydroxyproline, before and after 1, 3, 6, and 12 months; and serum calcium and phosphate monthly. During 12 months of the study, a substantial reduction in BMD was observed in all examined regions in group II (-2.8 +/- 2.1%; p<0.01 in L2-L4, -2.4 +/- 2.0%; p<0.01 in femoral neck, and -1.9 +/- 1.4%; p<0.01 in total body), whereas in group I a slight increase of bone mineral was noted, however insignificant. The inhibition of bone resorption was accompanied by a marked decrease in serum hydroxyproline. No changes in parathyroid activity were noted nor any decrease in serum phosphate. The treatment had no influence on serum endogenous calcitonin; initial concentrations were elevated in 47% of patients. CONCLUSION: Intranasal salmon calcitonin: 1) has no influence on bone mineralization in dialysis patients with uremic hyperparathyroidism; 2) has no significant effect on serum phosphate concentration; 3) provided adequate calcium supplementation doesn't stimulate parathyroid glands; 4) has no influence on endogenous calcitonin secretion. PMID- 15526841 TI - [Changes in selected parameters of bone turnover resulting from use of different calcium concentrations in dialysate of patients treated with CAPD]. AB - Renal osteodystrophy is a common complication occurring in patients with end stage disease. Using dialysis solutions with different concentrations of calcium undoubtedly has an influence on calcium-phosphorus homeostasis and bone turnover. The aim of the study was to examine the influence of dialysate with calcium concentrations of Ca 1.75 mmol/l and Ca 1.25 mmol/l on selected parameters of bone turnover. The study was performed on 12 clinically stable patients treated with CAPD. After 24 weeks of the treatment with Ca 1.75 mmol/l solution the initial investigation was performed. The next step was to switch to dialysate with the Ca level of 1.25 mmol/V. After 2, 12, and 24 weeks of the treatment with solutions of low calcium level the concentrations of PTH, osteocalcin, total calcium, and ionized calcium, phosphorus were calculated. We have observed that the level of PTH and osteocalcin was increased significantly at the time of treatment with Ca 1.25 mmol/l solution. We have also observed a decrease in concentration of total and ionized calcium. We did not observe any change of phosphorus concentrations in the serum of treated patients. PMID- 15526842 TI - [Angiotensin-converting enzyme gene polymorphism in patients with obstructive sleep apnea]. AB - Obstructive sleep apnea (OSA) is often accompanied by cardiovascular and metabolic disorders. The renin-angiotensin system plays an important role of the cardiovascular regulation. The activity of the angiotensin converting enzyme is genetically determined. The aim of the study was to estimate the relation between angiotensin converting gene polymorphism and cardiovascular diseases or familial history in patients with obstructive sleep apnea. 63 patients with OSA were enrolled to the study. Arterial hypertension was diagnosed in 30 cases, ischaemic heart disease in 5 cases and 10 patients suffered from both mentioned diseases. RESULT: The observed ACE genotype frequencies in the study group were II: ID: DD: 23.81: 47.62: 28.57%, in the hypertension subgroup II: ID: DD--30.0: 46.6: 23.3%. No differences in the study group were observed in relation to familial history :DI:II--30.43: 43.48: 26.09% vs 25.64: 48.72: 25.4%. CONCLUSIONS: 1. In the study group there was no association between ACE polymorphism and OSA. 2. There were also no association between the polymorphism of ACE and cardiac diseases or familial history of cardiac diseases in OSA group. 3. The polymorphism of ACE is not a risk factor for OSA. PMID- 15526843 TI - [Clinical usefulness of determining beta-2-microglobulin concentration in thyroid cystic fluid]. AB - The main subject of the paper was to prove that beta-2-microglobulin (beta-2-m) exists in fluid taken from a thyroid gland cyst of the patients with non-toxic, nodular goitre with cysts and whether the confirmation of beta-2-m existence in fluid taken from a cyst may indicate the presence of autoimmunological process in thyroid glands. The research was carried out on a group of 36 patients operated on because of goitre. Patients with the thyroid cancer were except. The operations were carried out in the General Surgery Department of Hospital 1 in Sosnowiec from 1994 to 1999. The group of patients consisted of women aged between 13 to 75 years old (average 43 +/- 13.8 years). TSH, FT4 serum concentration in this investigated group were normal. The paper presents the beta 2-m existence in fluid taken from a thyroid gland cyst. It was confirmed that there was no differences between beta-2-m serum concentration in operated patients and in healthy controls. In 50% of patients operated on with thyroid cysts high antibody antithyroid peroxidase (antiTPO) serum concentration and in fluid taken from cyst was confirmed and statistically higher beta-2-m concentration in cyst fluid than beta-2-m concentration in serum. CONCLUSIONS: The increased antiTPO concentration in serum and in a fluid taken from cyst and beta-2-m existence in cyst fluid point to the autoimmunological process taking place in half of the patients with thyroid cysts. PMID- 15526844 TI - [Changes of the oral mucous membrane in patients after transplantation of haematopoietic cells]. AB - Myeloablative chemotherapy used prior stem cell transplantation produces epithelial injury including oral cavity. Both damage by chemotherapy and subsequent infections cause mucositis. The aim of the study was to evaluate mucositis in a group of 31 patients. Patients aged 18-67 yrs (mean 42 yrs): 28 with blood neoplasms and 3 with aplastic anemia, were observed on days 0, 3, 5, 7, 9, 14 and 30 after transplantation of haematopoietic cells (autologous in 20 cases and allogeneic in 11 cases). The intensity of mucosal inflammation was evaluated according to 3 scales: WHO, Bearmann and 5-degree scale developed by the authors. This last scale, unlike other scales, was based on an accurate stomatological evaluation of the mucosal changes. A positive correlation was found between the intensity of mucosal inflammation and granulocytopenia <500/microl (p<0.01) and also between the presence of petechiae with blood platelet level <20.000/microl (p<0.001). PMID- 15526845 TI - [Use of argon plasma coagulation in advanced cancers of the upper digestive tract]. AB - Endoscopy being a diagnostic tool, allows also a nonsurgical treatment of upper gastrointestinal tract neoplasms, even at their advanced stage. Argon plasma coagulation (APC) is a contact-free method of monopolar coagulation, based primarily on the effects of high frequency electrical current flow in the ionized argon. The study used in 14 patients, aged 56-85 years, with advanced carcinoma of oesophagus and/or cardia. Carcinoma was diagnosed upon the results of endoscopic and microscopic examinations. The following parameters were included in the analysis: electrical current power, number of treatment sessions, stage of dysphagia, oesophagus passage, localization and type of a tumor. The analysis revealed that APC is a highly effective as well as safe method of palliative treatment of advanced oesophageal and/or cardiac cancers. Moreover, it was noted that APC is more effective in adenocarcinomas than in squamous cell carcinomas. PMID- 15526846 TI - [Outcome of immunosuppressive treatment of a patient with renal failure due to retroperitoneal fibrosis]. AB - Retroperitoneal fibrosis (RPF) is an uncommon disease of unclear aetiology. The review of the literature over the past 20 years revealed 160 published cases. Till now, no accepted diagnostic or therapeutic strategy exist. Most of patients diseases progress to end-stage renal failure without pharmacological treatment. In the paper we report a case of a 58-year old man admitted to the department of urology due to body mass loss, lower-back pain, vomiting, development of oliguria and anuria and intermittent claudication. On physical examination arterial hypertension (180/100 mm Hg), peripheral oedema, tenderness of the enlarged kidneys and lower limbs ischemia were found. Creatinine serum level (Pcr) was 232 micromol/l (2.69 mg/dl). On ultrasonography, symmetrical hydronephrosis and the existence of a hypoechogenic mass along of the aorta and below of renal arteries was found. The diagnosis of RPF was confirmed with MRI. Ureteral catheters were inserted with subsequent decompression of both kidneys and the patient was discharged from the hospital. Seven months later he still presented symptoms of lower limbs ischemia, arterial pressure was high and Per decreased to 138 micromol/l (1.55 mg/dl). The patient was admitted to the department of internal diseases. The treatment with prednisone at the dose of 40 mg/d during 6 weeks was introduced, and the dose was decreased gradually to 10 mg/d within 6 months. Simultaneously, the patient received intravenous therapy with cyclophosphamide 600 mg/infusion once monthly during 6 months. Two month after starting immunosuppressive treatment the intermittent claudication disappeared and after six months MRI examination demonstrated the regression of RPF. The ureteral catheters were removed. After 18 months of follow up, no recurrence of RPF is observed and the kidney function is normal. PMID- 15526847 TI - [Primary clear cell carcinoma of the thyroid--case report]. AB - We present a case of a 59-year old woman in whom incidentally a thyroid nodule of 10 mm diameter was discovered. A fine needle aspiration biopsy was performed and it revealed follicular tumour. The patient was referred to surgery and she had a subtotal thyreoidectomy performed with a part of the left lobe spared. The histopathological examination of the nodule led to a different diagnosis. The diagnosis was clear cell carcinoma in the thyroid, probably metastatic, the most probable primary site being the kidney. A computed tomography scan of the kidneys and abdomen was performed but it revealed no changes. The patient has been seen for a year in follow-up, with CT scans performed every six months that revealed no changes. The patient has been in good condition during the whole period of observation. CONCLUSIONS: We describe an extremely rare case of a primary thyroid clear cell carcinoma. PMID- 15526848 TI - [Pancreatic islets endocrinopathies with disorders of carbohydrate metabolism- Part 2: somatostatinoma, VIP-oma, PP-oma]. PMID- 15526849 TI - [The role of heparin in bronchial asthma]. PMID- 15526850 TI - [Obesity. The role of recently discovered hormones in the regulation of energy balance]. PMID- 15526851 TI - [Zinc supplementation in dialyzed patients]. PMID- 15526852 TI - [Patomechanism of diabetic macroangiopathy. The role of calcium-sensing receptor (CaR)]. PMID- 15526853 TI - [Genetic factors in the pathogenesis of atrial fibrillation]. PMID- 15526854 TI - [Assessment and monitoring of patients with chronic hepatitis C on antiviral therapy--state of the art in 2003]. PMID- 15526855 TI - Assessment of the genetic toxicological impacts of the Exxon Valdez oil spill on pink salmon (Oncorhynchus gorbuscha) may be confounded by the influence of hatchery fish. PMID- 15526856 TI - Integration of ecotoxicity index and carboxylic of MSW incineration bottom ashes. AB - In France, MSW incineration bottom ash disposal calls for physico-chemical tests and depending on the results the bottom ashes are sorted into different classes allowing the reuse or not. However this study shows that there is no direct correlation between the physico-chemical testings, like leaching, soluble fraction and heavy metal content, and the environmental impact, in terms of biological toxicity. Based on the results of several ecotoxicity testings and their biotest response we define an "ecotoxicity index" and demonstrate there is a correlation between the carboxylic acids amount and the ecotoxicologic index for the bottom ashes. This clearly shows that at least organic carbon characterisation in the bottom ash is needed in addition to leaching and physico chemical testings if one wants to have a coherent sorting procedure for the bottom ashes. PMID- 15526857 TI - Experimental designs to assess endocrine disrupting effects in invertebrates. A review. AB - In order to gain basic understanding of the ecological effects of vertebrate Endocrine Disrupting Chemicals (EDCs), many research groups are currently testing these chemicals using aquatic invertebrates. Small crustaceans, such as cladocerans and copepods, are of particular interest since they are ecologically important and their short life cycles allow obtaining information on demographic parameters. Despite the existence of diverse literature on the development, growth and reproductive effects of EDCs on these crustaceans, only a few studies have unambiguously assessed a truly endocrine disrupting effect. This review discusses new experimental designs to differentiate between endocrine disruption and other causes of reproductive and developmental impairment. Our findings clearly illustrate that many studies may have falsely concluded that chemicals have endocrine disrupting modes of action when in fact a much simpler explanation was not previously ruled out (e.g., egg mortality, feeding inhibition). This means that there is an urgent need for integration of toxic effects on energy intake to toxicity assessments. Such an approach would permit different ectotoxicological models of action, including endocrine disrupting effects, to be distinguished and their relative roles in the overall toxic response to be clarified. PMID- 15526859 TI - Effects of mercury on the life table demography of the rotifer Brachionus calyciflorus Pallas (Rotifera). AB - Mercury is highly toxic to a variety of aquatic organisms including zooplankton. The functioning of freshwater ecosystems can be altered if rotifers, being a natural food link between phytoplankton and fish larvae, are contaminated by mercuric compounds. In order to detect age-specific responses of the rotifer Brachionus calyciflorus to mercury toxicity (5 nominal concentrations as chloride viz. 0, 0.000625, 0.00125, 0.0025 and 0.005 mg l(-1)), we used the standard life table method at two different food (Chlorella vulgaris) levels (0.5 x 10(6) and 1.5 x 10(6) cells ml(-1)). Data indicated that increase in mercury concentration had an increasingly intense negative effect on many of the life history variables, while at higher food levels, its impact was less. A nearly rectangular survivorship pattern was obtained in controls, especially at higher food levels. This trend gradually changed to a steep fall as the concentration of the heavy metal in the medium increased from 0 to 0.005 mg l(-1). At any given food density, increase in the mercury concentration resulted in decreased age-specific reproduction. A maximum of 3.5 offspring female(-1) was observed in controls at higher food density. The average lifespan varied from 6 to 8 days at low food level, depending on the heavy metal concentration in the medium. The corresponding values at high food level varied from 8 to 12 days. Regardless of mercury concentration in the medium, gross and net reproductive values varied from 10 to 33 and 4 to 19 offspring female(-1). The longest generation time (about 9 days) of B. calyciflorus was obtained at 1.5 x 10(6) cells ml(-1) food density in control, while the shortest was 5 days at low food level and high (0.005 mg l(-1)) mercury concentration in the medium. Depending on the food level and heavy metal concentration in the medium, the rate of population growth (r) varied from 0.32 to 0.62 d(-1). In general, higher food level resulted in higher r. Except generation time, all other derived variables were significantly influenced by food level and the heavy metal concentration in the medium. PMID- 15526858 TI - Tissue distribution and redistribution of trace elements in shrimp species with the emphasis on the roles of metallothionein. AB - This review is focused on distribution and redistribution of trace elements in various tissues of different shrimp species, especially genus Penaeus. The possible roles of metallothionein in this regard are emphasized. Factors affecting heavy metals uptake and distribution have also been reviewed separately. Moreover, patterns of metals bioaccumulation and their order of occurrence have been evaluated. Another part of this paper deals with comparison of the related data from different aquatic environments as well as existing guidelines and limits for human consumption. Generally, the number of conducted studies in some related context, especially in the case of possible roles of metallothionein during storage of organisms is very limited. PMID- 15526860 TI - Use of fatty acids to explain variability of organochlorine concentrations in eggs and plasma of common terns (Sterna hirundo). AB - We have studied the breeding parameters, organochlorine compounds (OCs) concentrations, and fatty acid (FA) composition of egg yolks (n = 47) and plasma (n = 90) of common terns (Sterna hirundo) from two colonies (Banya and Fangar) in the Ebro delta, NE Spain. Terns from Banya tend to have smaller clutch size and lower hatching success than terns from Fangar. p,p'-DDE and polychlorinated biphenyls (PCBs) concentrations were almost 2-fold higher in yolks from Banya in 1998 than from Fangar in 1999, and the percentage of n-6 PUFA was positively correlated with these contaminants. Differences between samplings in OCs concentrations in plasma were less evident, and were affected by breeding chronology. The highest OCs concentrations in yolks from Banya may be explained by two processes involving the increased deposition of n-6 PUFA: (1) higher mobilization of endogenous fat due to lower food resources, or (2) differences in the diet between colonies. Birds from Banya may have been feeding at a higher degree on discards of trawling fisheries composed of demersal and benthic fish species that are more exposed to contaminants from sediment and have lower n-3/n 6 ratio, whereas birds from Fangar would feed mostly on pelagic species of small clupeiformes that are less polluted and have higher n-3/n-6 ratios. PMID- 15526861 TI - Biodiversity of Collembola in urban soils and the use of Folsomia candida to assess soil 'quality'. AB - The effects of metal contamination on natural populations of Collembola in soils from five sites in the Wolverhampton area (West Midlands, England) were examined. Analysis revealed that metal concentrations were elevated above background levels at all sites. One location in particular (Ladymoor, a former smelting site) was highly contaminated with Cd, Cu, Pb and Zn at more than 20 times background levels. Biodiversity indices (Shannon-Weiner, Simpson index, Margalef index, alpha index, species richness, Shaneven (evenness) and Berger-Parker dominance) were calculated. Of these indices, estimates of species richness and evenness were most effective at highlighting the differences between the Collembola communities. Indeed, the highest number of species were found at the most contaminated site, although the Collembola population also had a comparatively low evenness value, with just two species dominating. The number of individuals per species were allocated into geometric classes and plotted against the cumulative number of species as a percentage. At Ladymoor, there were more geometric classes, and the slope of the line was shallower than at the other four sites. This characteristic is a feature of polluted sites, where a few species are dominant and most species are rare. The Ladymoor soil also had a dominance of Isotomurus palustris, and was the only site in which Ceratophysella denticulata was found. Previous studies have shown that these two species are often found in sites subject to high metal contamination. Survival and reproduction of the "standard" test springtail, Folsomia candida (Willem), were determined in a 4 week exposure test to soils from all five sites. Mortality was significantly increased in adults and reproduction significantly lower in the Ladymoor soil in comparison to the other four sites. This study has shown that severe metal contamination can be related to the population structure of Collembola in the field, and performance of F. candida (in soils from such sites) in the laboratory. PMID- 15526862 TI - A comparative study of the effects of metal contamination on Collembola in the field and in the laboratory. AB - We examined the species diversity and abundance of Collembola at 32 sampling points along a gradient of metal contamination in a rough grassland site (Wolverhampton, England), formerly used for the disposal of metal-rich smelting waste. Differences in the concentrations of Cd, Cu, Pb and Zn between the least and most contaminated part of the 35 metre transect were more than one order of magnitude. A gradient of Zn concentrations from 597 to 9080 microg g(-1) dry soil was found. A comparison between field concentrations of the four metals and previous studies on their relative toxicities to Collembola, suggested that Zn is likely to be responsible for any ecotoxicological effects on springtails at this site. Euedaphic (soil dwelling) Collembola were extracted by placing soil cores into Tullgren funnels and epedaphic (surface dwelling) species were sampled using pitfall traps. There was no obvious relationship between the total abundance, or a range of commonly used diversity indices, and Zn levels in soils. However, individual species showed considerable differences in abundance. Metal "tolerant" (e.g., Ceratophysella denticulata) and metal "sensitive" (e.g., Cryptopygus thermophilus) species could be identified. Epedaphic species appeared to be influenced less by metal contamination than euedaphic species. This difference is probably due to the higher mobility and lower contact with the soil pore water of epedaphic springtails in comparison to euedaphic Collembola. In an experiment exposing the standard test springtail, Folsomia candida, to soils from all 32 sampling points, adult survival and reproduction showed small but significant negative relationships with total Zn concentrations. Nevertheless, juveniles were still produced from eggs laid by females in the most contaminated soils with 9080 microg g(-1) Zn. Folsomia candida is much more sensitive to equivalent concentrations of Zn in the standard OECD soil. Thus, care should be taken in extrapolating the results of laboratory toxicity tests on metals in OECD soil to field soils, in which, the biological availability of contaminants is likely to be lower. Our studies have shown the importance of ecotoxicological effects at the species level. Although there may be no differences in overall abundance, sensitive species that are numerous in contaminated sites, and which may play important roles in decomposition ("keystone species") can be greatly reduced in numbers by pollution. PMID- 15526863 TI - Coumaphos distribution in the hive ecosystem: case study for modeling applications. AB - Pesticides are currently used inside hives, against the honeybee parasite Varroa destructor, producing unwanted contamination effects. To assess the distribution and fate of one of these pesticides (coumaphos), two experimental hives were treated with Perizin (the commercial product containing the active ingredient coumaphos). Samples of honey, wax, pollen, adult bees and larvae taken before treatment and up to 104 days afterwards, showed diffuse contamination. Wood hedges and wax bridges, where the pesticide solution was applied, were analysed as well. A mass balance was calculated, yielding a recovered amount of around 60% just after treatment and 38% 1 month later. Directly contaminated surfaces and wax contained the highest amount of residues. Wax and honey contained different amounts (10, and 0.1% respectively) but both retained residues for long time. Bees ingest most of the product just after treatment, then rapidly eliminate it by metabolism, advection and deposition processes. On the basis of analytical results, a simple model (level I of the fugacity model) was applied to the hive system for different pesticides (coumaphos, malathion, fluvalinate and bromopropylate). Predicted concentrations in wax and honey were compared with those measured, indicating the good predictive capability of this approach. PMID- 15526864 TI - Assessment of soil toxicity from an antitank firing range using Lumbricus terrestris and Eisenia andrei in mesocosms and laboratory studies. AB - Earthworm mesocosms studies were carried out on a explosives-contaminated site at an antitank firing range. Survival of earthworms and the lysosomal neutral red retention time (NRRT), a biomarker of lysosomal membrane stability, were used in these studies to assess the effect of explosives-contaminated soils on the earthworms Lumbricus terrestris and Eisenia andrei under field conditions. Toxicity of the soils samples for E. andrei was also assessed under laboratory conditions using the earthworms reproduction test and the NRRT. Results indicate that the survival was reduced up to 40% in certain explosive-contaminated soil mesocosms following 10 days of exposure under field conditions, whereas survival was reduced up to 100% following 28 days of exposure under laboratory conditions. Reproduction parameters such as number of cocoons and number of juveniles were reduced in many of the selected contaminated soils. Compared to the reference, NRRT was significantly reduced for E. andrei exposed to explosive-contaminated soils under both field and laboratory conditions, whereas for L. terrestris NRRT was similar compared to the reference mesocosm. Analyses showed that HMX was the major polynitro-organic compound in soils. HMX was also the only explosive detected in earthworm tissues. Thus, results from both field mesocosms and laboratory studies, showed lethal and sub-lethal effects associated to soil from the contaminated area of the antitank firing range. PMID- 15526865 TI - Suffering of gravely ill patients. An important area of intervention for family physicians. PMID- 15526866 TI - Reinventing primary health care. Physicians have a pivotal part to play. PMID- 15526867 TI - More thoughts on third-year training. PMID- 15526868 TI - More thoughts on third-year training. PMID- 15526869 TI - [More thoughts on third-year training]. PMID- 15526870 TI - Home care critical part of health care. PMID- 15526871 TI - Celiac disease during pregnancy. AB - QUESTION: One of my patients was diagnosed with celiac disease and maintains a strict gluten-free diet. Is her fetus at risk of neural tube defects because she does not get folic acid from bread and other flour-based foods? ANSWER: A woman with celiac disease must supplement her diet with multivitamins, including folic acid. Most prenatal vitamins contain 0.8 to 0.9 mg of folic acid, double the amount recommended by Health Canada for prevention of neural tube defects. Without supplementation (eg, undiagnosed pregnancy), women with celiac disease might not take in enough folate to maintain protective levels. PMID- 15526872 TI - Ophthaproblem. Charles Bonnet's syndrome. PMID- 15526873 TI - Practice tips. Website for your family practice. PMID- 15526874 TI - An approach to managing depression. Defining and measuring outcomes. AB - OBJECTIVE: To provide family physicians with a contemporary approach to formulating a treatment model for major depressive disorder that integrates definitions of new therapeutic end points, familiarizes them with tools for assessing these end points, and describes newer methods for enhancing outcome. SOURCES OF INFORMATION: Canadian Psychiatric Association Guidelines for the Treatment of Depressive Disorders, relevant articles from a MEDLINE search using the MeSH headings"full remission" and "depression," and the authors' clinical experience. MAIN MESSAGE: Major depressive disorder is an episodic, relapsing, and sometimes chronic illness. Depressive symptoms in primary care settings are often vague reports of anhedonia, anxiety, and nonspecific somatic complaints. Therapeutic objectives in depression are full remission of depressive symptoms, prevention of recurrence, and restoration of function. Depression rating scales can be useful for monitoring and treating depression. CONCLUSION: The proposed therapeutic model anticipates the chronic course of illness, defines treatment end points, encourages longer duration of treatment, and includes both pharmacologic and lifestyle therapies. The 7-item Hamilton Depression Rating Scale can assist clinicians in determining when full remission has occurred. PMID- 15526875 TI - Case report: adverse drug reactions in unrecognized kidney failure. PMID- 15526876 TI - Women marginalized by poverty and violence. How patient-physician relationships can help. AB - OBJECTIVE: To explore the experiences of women relegated to the margins of society by poverty or violence. DESIGN: Qualitative method of focus groups. SETTING: Shelters and transitional housing in southwestern Ontario. PARTICIPANTS: Thirty-six women staying at shelters or transitional housing. METHOD: Focus groups conducted at five locations explored the women's experiences and interactions with family physicians. MAIN FINDINGS: Two themes emerged from the analysis: power imbalances in patient-physician relationships, and the role of family physicians in creating collaborative relationships. Women who felt demeaned in patient-physician relationships described their family physicians as dominating and intimidating. Women who described relationships as collaborative felt valued and understood. CONCLUSION: Poor or abused women living in shelters who felt powerless in patient-physician relationships felt even more demeaned as they coped with the struggles associated with being poor. Women who had continuous collaborative relationships with their family physicians were able to articulate their needs more readily. PMID- 15526877 TI - Evidence-based cardiovascular care. Family physicians' views of obstacles and opportunities. AB - OBJECTIVE: To explore obstacles to and opportunities for applying specific lifestyle and pharmacologic recommendations on chronic ischemic heart disease. DESIGN: Qualitative study. SETTING: Rural, town, and city settings in Nova Scotia. PARTICIPANTS: Fifty family physicians caring for patients with cardiovascular (CV) disease. METHOD: Nine focus groups were conducted, audiotaped, and transcribed. Seven recommendations had been selected for discussion based on their relevance to primary care, strength, and class of supporting evidence. Analysis was guided by grounded-theory methodology. MAIN FINDINGS: "Ischemic events" can be powerful motivators for change, whereas the asymptomatic nature of CV risks and distant outcomes can form obstacles. Trust built through previous experiences and the opportunity to repeat important messages can facilitate application of evidence, but patient-physician relationships can also pose obstacles. CONCLUSION: Physicians can take steps to improve care, but success at reducing CV risks depends upon active involvement of many health professionals and community resources. Future guideline implementation should focus on patient-oriented issues, such as comorbidity and treatment preferences. PMID- 15526878 TI - Short report: Ontario family medicine residents. Practice choices in 1998 and 1999. PMID- 15526879 TI - Canadian Hypertension Education Program. Brief overview of 2004 recommendations. PMID- 15526880 TI - Adverse drug reactions in Canada. PMID- 15526881 TI - Effects of porcine zona pellucida immunocontraceptives in zoo felids. AB - Methods of contraception are necessary for management of zoo felids; however, the most commonly used contraceptive (melengestrol acetate implant) is associated with serious adverse reactions with long-term use. Porcine zona pellucida (pZP) vaccines are promising as contraceptives, but their safety in zoo felids has not been tested. pZP vaccine was administered to 27 female felids representing 10 species, including African lion (Panthera leo), Asian leopard (P. pardus), jaguar (P. onca), tiger (P. tigris), snow leopard (P. uncia), cougar (Felis concolor), Siberian lynx (F. lynx), Canada lynx (F. canadensis), serval (F. serval), and bobcat (F. rufus), in 15 facilities. Over 6 wk, each animal received three i.m. injections of 65 microg pZP with Freund's complete adjuvant (FCA), Freund's incomplete adjuvant, or carbopol as the adjuvant. Behavioral signs of estrus were seen in 14 of the vaccinated felids. An unacceptably high incidence of adverse reactions was seen including injection site swelling, lameness, limb swelling, or abscessation (or all) in five felids after injection with FCA as the initial adjuvant. Adverse behavioral signs, including increased irritability and aggression, were seen in four felids. Six of the felids were assayed for antibodies against pZP during the 12 mo after vaccination; all showed antibody production. Antibody levels appeared to peak 1-4 mo after vaccination began, although elevated antibody levels persisted in two animals for > 12 mo after the first injection. All vaccinated felids were ovariohysterectomized 3-13 mo after vaccination. Folliculogenesis was present in all treated animals, and there was no histopathologic evidence of inflammatory damage to ovaries. Contraceptive efficacy was not specifically evaluated in this study; however, two of the three felids housed with an intact male became pregnant during the study, one of which gave birth to healthy cubs. PMID- 15526882 TI - Pancreatic islet fibrosis in rock hyrax (Procavia capensis), Part 2: Pathology, immunohistochemistry, and electron microscopy. AB - Pancreatic islet fibrosis with varying degrees of islet cell hyperplasia or islet effacement was diagnosed histologically in 19 rock hyraxes (Procavia capensis) from seven zoological parks. Some, but not all, affected hyraxes were from a common lineage. The condition was associated with apparent hyperglycemia in seven and diabetes mellitus in two. Immunohistochemistry revealed hyperplasia of beta, alpha, and delta cells proportional to the degree of the fibrosis. Electron microscopy revealed collagen deposition and fibroplasia within and around the islets. Special stains and electron microscopy were negative for the presence of amyloid. Beta cell depletion was never identified. The condition has morphologic features that resemble islet fibrosis of human infants born to diabetic mothers. PMID- 15526883 TI - Retrospective evaluation of renal disease in captive black howler monkeys (Alouatta caraya). AB - Six of 15 (40%) inactive medical records of adult black howler monkeys (Alouatta caraya) at one zoological institution included either a pre- or postmortem diagnosis of renal disease. In these six cases, significantly abnormal hematologic and serum chemistry values were reported at onset of azotemia, onset of clinical signs, and at euthanasia. Average age of onset of azotemia was 14.8 +/- 2.9 yr, with clinical signs of disease noted at 17 +/- 4.7 yr. In four of the cases (66.6%), azotemia was documented earlier than the onset of clinical signs of renal disease. Average duration of clinical disease was 2.83 +/- 1.6 yr, with an average age at euthanasia of 18 +/- 4.7 yr. Chronic tubulointerstitial nephritis with secondary glomerular sclerosis was present in all cases. Thirteen of an additional 20 institutions in the United States that have held Alouatta caraya responded to a survey for prevalence of renal disease. These institutions showed a lower prevalence (15.1%) of renal disease in complete, inactive records, a higher prevalence of glomerulonephritis, and similar significant clinicopathologic values. PMID- 15526884 TI - Field use of isoflurane and air anesthetic equipment in wildlife. AB - Conventional inhalation anesthesia of wildlife species within natural habitats presents significant practical problems. Heavy cylinders of medical grade oxygen are often unavailable in field situations. Equipment has been modified to permit the delivery of isoflurane in ambient air as the carrier and to be fitted with circuitry adaptable for different species and anesthetic situations. Preliminary empirical studies at low altitude in a range of small mammalian and avian species demonstrate the suitability of this combination and these techniques for inducing and maintaining anesthesia in clinically normal patients undergoing relatively minor procedures. The equipment has also been used to deepen and prolong anesthesia in several larger species, including great apes and large cats, after induction with injectable agents. These techniques, in combination with pulse oximetry to detect hypoxemia, provide a cheap, robust, and portable inhalation anesthetic system for field situations that is not dependent on compressed gasses. PMID- 15526885 TI - Chemical immobilization of rhebok (Pelea capreolus) with carfentanil-xylazine or etorphine-xylazine. AB - Twelve adult rhebok (Pelea capreolus) were immobilized using a combination of 0.4 mg/kg xylazine and either 0.01 mg/kg of carfentanil (n = 6) or 0.01 mg/kg etorphine (n = 6), delivered i.m. using a remote injection system. Induction and recovery times, heart rate, respiratory rate, rectal temperature, oxygen saturation, end-tidal CO2 (ETCO2), anesthetic depth, indirect blood pressure, and arterial blood gases were recorded. Rhebok were not intubated but nasal oxygen was administered. Forty minutes after induction, anesthesia was antagonized with naltrexone and yohimbine. Mean initial heart rate was significantly higher in the carfentanil group than in the etorphine group. Mean initial oxygen saturation was consistent with hypoxia in both the carfentanil group and the etorphine group. In both groups, arterial pH decreased and partial pressure of carbon dioxide increased during the first 15 min of anesthesia, and values were similar in both groups. These findings were consistent with respiratory acidosis and decreased ventilation. Values for respiratory rate, temperature, oxygen saturation, ETCO2, and blood pressure were similar for both groups at all time periods. During the first 5 min of anesthesia, rhebok in the carfentanil group were more responsive to stimuli than rhebok in the etorphine group. After administration of antagonists, time to first arousal was significantly shorter in the etorphine group than in the carfentanil group. Although cardiopulmonary values were similar for the two groups, rhebok in the carfentanil group were at a comparatively lighter plane of anesthesia, and some individuals in this group required additional manual and chemical restraint for medical procedures to be performed. In conclusion, for captive adult rhebok, 0.01 mg/kg of etorphine and 0.4 mg/kg of xylazine are recommended over 0.01 mg/kg carfentanil and 0.4 mg/kg xylazine because of qualitatively better anesthetic episodes and shorter recovery times. PMID- 15526887 TI - Comparison of the effect of dipotassium ethylenediaminetetraacetic acid and lithium heparin on hematologic values in the green iguana (Iguana iguana). AB - We compared the effects of dipotassium ethylenediaminetetraacetic acid (EDTA) and lithium heparin on hematologic values of green iguanas (Iguana iguana). Thirty two privately owned sibling iguanas had blood drawn, and the sample was divided into three components: an EDTA tube, a heparin tube, and a nonanticoagulated blood smear. A full reptilian complete blood count was performed on each anticoagulated sample, and white blood cell (WBC) and leukocyte differential counts were performed on the whole-blood smears. Heparin and EDTA samples differed significantly in absolute values of thrombocytes, WBC, heterophils, and monocytes. The EDTA had no significant effect on the packed cell volume or plasma protein values, and the white blood count and differential counts produced with EDTA were more similar to those of the nonanticoagulated blood smear than were the counts produced with heparin. PMID- 15526886 TI - Xylazine-midazolam-ketamine versus medetomidine-midazolam-ketamine anesthesia in captive Siberian tigers (Panthera tigris altaica). AB - Two alpha2-adrenoceptor agents, xylazine and medetomidine, in combination with midazolam and ketamine safely and effectively immobilized Siberian tigers (Panthera tigris altaica). The medetomidine protocol used smaller drug volumes, and induction and recovery times were shorter. Although cardiopulmonary abnormalities were noted, none were likely to be life threatening. PMID- 15526888 TI - Pharmacokinetics of enrofloxacin after single-dose oral and intravenous administration in the American alligator (Alligator mississippiensis). AB - The pharmacokinetics of enrofloxacin administered orally and i.v. to American alligators (Alligator mississippiensis) at 5 mg/kg was determined. Plasma levels of enrofloxacin and its metabolite ciprofloxacin were measured using high performance liquid chromatography and the resulting concentration versus time curve analyzed using compartmental modeling techniques for the i.v. data and noncompartmental modeling techniques for the oral data. A two-compartment model best represented the i.v. data. Intravenous administration of enrofloxacin resulted in an extrapolated mean plasma concentration of 4.19 +/- 4.23 microg/ml at time zero, with average plasma drug levels remaining above 1.0 microg/ml for an average of 36 hr. Plasma volume of distribution for i.v. enrofloxacin was 1.88 +/- 0.96 L/kg, with a harmonic mean elimination half-life of 21.05 hr and mean total body clearance rate of 0.047 +/- 0.021 L/hr/kg. Plasma levels of p.o. enrofloxacin remained below 1.0 microg/ml in all test animals, and average concentrations ranged from 0.08 to 0.50 microg/ml throughout the sampling period. Oral administration of enrofloxacin achieved a mean maximum plasma concentration of 0.50 +/- 0.27 microg/ml at 55 +/- 29 hr after administration, with a harmonic mean terminal elimination half-life of 77.73 hr. Minimal levels of ciprofloxacin were detected after both oral and i.v. enrofloxacin administration, with concentrations below minimum inhibitory concentrations for most susceptible organisms. On the basis of the results of this study, enrofloxacin administered to American alligators at 5 mg/kg i.v. q 36 hr is expected to maintain plasma concentrations that approximate the minimum inhibitory concentration for susceptible organisms (0.5 microg/ml). Enrofloxacin administered to American alligators at 5 mg/kg p.o. is not expected to achieve minimum inhibitory values for susceptible organisms. PMID- 15526889 TI - Pharmacokinetic disposition of a long-acting oxytetracycline formulation after single-dose intravenous and intramuscular administrations in the American alligator (Alligator mississippiensis). AB - The pharmacokinetics of a long-acting oxytetracycline preparation administered i.v. and i.m. to American alligators (Alligator mississippiensis) at 10 mg/kg was determined. Plasma levels of oxytetracycline were measured using high-performance liquid chromatography, and the resulting concentration versus time curve was analyzed using compartmental modeling and noncompartmental modeling techniques for i.v. and i.m. samples, respectively. A two-compartment model best represented the i.v. data. Intravenous administration of oxytetracycline resulted in an extrapolated mean plasma concentration at time zero of 60.63 +/- 28.26 microg/ml, with average plasma drug levels of 2.82 +/- 0.71 microg/ml at the end of the 192 hr sampling period. Plasma volume of distribution for i.v. oxytetracycline was 0.20 +/- 0.09 L/kg, with a harmonic mean elimination half-life of 15.15 hr and mean total body clearance rate of 0.007 +/- 0.002 L/hr/kg. Intramuscular administration of oxytetracycline achieved a mean peak plasma concentration of 6.85 +/- 1.96 microg/ml at 1 hr after administration, with average plasma drug levels of 4.96 +/- 1.97 microg/ml at the end of the 192-hr sampling period. The harmonic mean terminal elimination half-life for i.m. oxytetracycline was 131.23 hr. Based on the results of this study, long-acting preparations of oxytetracycline administered parenterally to American alligators at 10 mg/kg q 5 days is expected to maintain plasma concentrations above the minimum inhibitory concentration of 4.0 microg/ml for susceptible organisms. PMID- 15526890 TI - Effects of ozonation on the speciation of dissolved iodine in artificial seawater. AB - Iodine in the form of iodide is required for synthesis of tri-iodothyronine and thyroxine in fish. Iodine chemical speciation in aliquots of raw artificial seawater mix was measured before, during, and after exposure for fixed time periods to air only and to concentrations of ozone required to achieve oxidation reduction potentials typical of a protein skimmer (400 mV) and an ozone contact chamber (800 mV). Chemical species of iodine were also measured in tank water from a large, recirculating, ozonated aquarium system that has a low-grade incidence of thyroid lesions (e.g., thyroiditis, hyperplasia, adenoma, and adenocarcinoma) in its fish. With increasing exposure to ozone, concentrations of iodide and dissolved organic iodine (DOI) decreased, whereas iodate levels increased. As a result of exposure to 400 mV, iodide concentration dropped to less than half the amount found in raw artificial seawater mix. After exposure to 800 mV, initial iodide levels decreased by 67%, and DOI became undetectable, whereas iodate concentration increased by 155%, with no remarkable change in total iodine concentration. These results indicate ozone-induced conversions from iodide to iodate, and DOI to iodide or iodate (or both). Iodide and DOI were not detectable in the aquarium system's water samples. Ozonation of artificial seawater may alter the relative concentrations of iodine species in a closed tank system, so that iodide supplementation of the diet or tank water of captive teleosts and elasmobranchs living in ozonated seawater is advisable. PMID- 15526891 TI - Renal ultrasonographic evaluation in the oncilla (Leopardus tigrinus). AB - Ultrasonography is currently used in veterinary medicine to examine the anatomy and physiology of internal organs and to establish normal standards for exotic animal species. Specifically, the kidneys may be evaluated and measured in length, width, and height. The kidneys in 33 oncilla (Leopardus tigrinus), including 20 males and 13 females, were opportunistically located, characterized, and measured by ultrasound using a 7.5-MHz linear transducer at two different zoologic facilities in Sao Paulo, Brazil. Renal volume was calculated from these linear measurements. The mean linear measurements and volume of the right and left kidneys were not significantly different. PMID- 15526892 TI - Pancreatic islet fibrosis in rock hyraxes (Procavia capensis), Part 1: Case histories, clinical pathology, and epizootiology. AB - Two adult female rock hyraxes (Procavia capensis) at the Dallas Zoo were confirmed with spontaneous diabetes mellitus from 1997-2000, whereas a third animal with a similar clinical presentation never became hyperglycemic. The pancreas in all three animals showed pancreatic islet fibrosis (PIF). Retrospective examination of medical records for rock hyraxes acquired by this collection or born into it from 1991-2002 identified eight more animals affected with PIE All affected animals, including three males and eight females, were 1-7 yr of age and presented either with vague clinical signs of soft feces and rough hair coat or were acutely moribund or dead. Clinical pathology data was available for seven of the animals before onset of overt clinical signs and revealed inappropriate hyperglycemia in six, as well as elevated serum concentrations of creatine phosphokinase, amylase, and lipase in all seven animals. Pedigree evaluation did not support a familial pattern for PIE Review of the histopathology findings from nine other zoologic collections with rock hyrax deaths during the study period identified six institutions with 12 additional cases genetically unrelated to the incident collection. Histopathology and viral serology did not support an infectious cause. Analysis of serum anti-islet and anti-insulin antibodies did not suggest autoimmune disease, and none of the animals had known exposure to toxic substances. Limited nutritional analyses did not support a nutritional basis for the condition, and the cause for PIF remains unknown. PMID- 15526893 TI - Transitional cell carcinomas in four fishing cats (Prionailurus viverrinus). AB - Transitional cell carcinomas (TCC) of the urinary bladder were diagnosed in four related fishing cats (Prionailurus viverrinus). The major clinical sign in each case was persistent hematuria unresponsive to medical therapy. Cystotomy and biopsy provided an antemortem diagnosis in three of the fishing cats before euthanasia because of progression of clinical signs. The diagnosis was made in the fourth cat after euthanasia because of renal failure. Hematuria improved temporarily in one of the cats diagnosed antemortem and treated with piroxicam and carboplatin. Attempts to isolate a herpesvirus in two of the cats failed. Histopathologic appearance of the TCC was similar to that described for other species. TCC metastasis to the lungs was noted at necropsy in one cat; metastatic disease was not noted in the other fishing cats on gross or histopathologic examination. TCC of the urinary bladder appears to be more prevalent in fishing cats than in other species of domestic or nondomestic felids. PMID- 15526894 TI - Renal myxosporidiasis in Asian horned frogs (Megophrys nasuta). AB - Renal myxosporidiasis and associated lesions were seen in Asian horned frogs, Megaphrys nasuta, at the Detroit Zoological Institute and the Toledo Zoo. The organism was identified as Chloromyxum sp. on the basis of histopathology, cytology, and electron microscopy. Histologic changes in the kidneys included varying degrees of renal tubular dilation and necrosis, and mild to severe nonsuppurative tubulointerstitial nephritis associated with vegetative stages of the myxosporidian. This is the second known identification of Chloromyxum sp. in amphibian kidneys, and the first report of death attributed primarily to the parasite. PMID- 15526895 TI - Serologic survey in a colony of captive common marmosets (Callithrix jacchus) after infection with herpes simplex type 1-like virus. AB - An outbreak of herpesvirus caused the death of four of five common marmosets (Callithrix jacchus) in a private colony. Gross lesions included acute ulcerative gingivitis, glossitis, and enlarged mandibular lymph nodes. Histologically, all fatal cases showed meningoencephalitis and eosinophilia with intranuclear inclusion bodies in neurons and glial cells. A herpes simplex-like virus was cultured from the brain and was identified as herpes simplex type 1 virus or a closely related virus by immunofluorescence. Serologic testing (complement fixation test) indicated that the surviving adult female was serologically positive for more than 4 yr and that the offspring she produced was seronegative. The most likely source of the outbreak was the owner who mouth fed hand-raised offspring. PMID- 15526896 TI - Use of computer tomography for imaging of Crassicauda grampicola in a Risso's dolphin (Grampus griseus). AB - A mature male Risso's dolphin (Grampus griseus) stranded along the coasts of Friuli Venezia Giulia, northeast Italy, in May 2001. Parasitic infection with Crassicauda grampicola is often found in the tympanic bullae and pterygoid sinuses in many of the Risso's dolphins examined from the same area. For this reason, it was decided to perform computed tomography of the head to assess this imaging technique for the diagnosis of crassicaudosis in dolphins. A full postmortem examination confirmed the pathologic findings of the computed tomography scan. This technique can be considered a useful adjunct in the diagnosis of cranial crassicaudosis in live dolphins. PMID- 15526897 TI - Plasma evaluation for ivermectin in llamas (Lama glama) after standard subcutaneous dosing. AB - Plasma levels of the parasiticide ivermectin were studied by high-performance liquid chromatography in five llamas (Lama glama) after single 200 microg/kg s.c. injections. Ivermectin levels were undetectable in plasma samples drawn up to 4 wk after injection, suggesting that the dosage used was insufficient to reach therapeutic concentrations in this species. PMID- 15526898 TI - Diagnosis and treatment of presumptive pyelonephritis in an Asian elephant (Elephas maximus). AB - A 37-yr-old female Asian elephant (Elephas maximus) presented with anorexia, restlessness, and dark-colored urine. Urinalyses showed hematuria, leukocyturia, isosthenuria, proteinuria, granular casts, and no calcium oxalate crystals. Bloodwork revealed azotemia. Urine culture revealed a pure growth of Streptococcus zooepidemicus resistant to sulfamethoxazole-trimethoprim but susceptible to cephalosporins. A presumptive diagnosis of pyelonephritis was made based on bloodwork, urinalysis, and urine culture. The animal was treated with intravenous ceftiofur, and intravenous and per rectum fluids were given for hydration. The elephant's attitude and appetite returned to normal, the abnormal blood parameters resolved, and urinary calcium oxalate crystals reappeared after treatment, supporting presumptive diagnosis. Follow-up ultrasonography revealed an abnormal outline of both kidneys with parenchymal hyperechogenicity and multiple uterine leiomyomas. PMID- 15526899 TI - Knemidokoptes in a Swainson's hawk, Buteo swainsoni. AB - A male Swainson's hawk (Buteo swainsoni) at a rehabilitation facility in Colorado was presented with severe crusting of the skin on the medial thighs and caudal ventral body. Male and female Knemidokoptes mites were found on a cellophane tape preparation of the affected area. The hawk's clinical signs resolved with treatment with ivermectin administered at 200 microg/kg, i.m., every 10 days for three treatments. PMID- 15526900 TI - Malformation of the permanent maxillary canine after dental infraction in a wild boar (Sus scrofa L.). AB - Intra-alveolar infraction and malformation of the apical part of the permanent left maxillary canine in an adult male wild boar (Sus scrofa L.) is documented. The infraction and its sequelae produced an irregular mass of reparative tissue that led to the formation of two growing ends of the tooth. PMID- 15526901 TI - Retroperitoneal teratoma in a skunk (Mephitis mephitis). AB - A 4-yr-old striped skunk (Mephitis mephitis) developed an 11- x 7- x 7-cm retroperitoneal mass caudal to the left kidney. The mass was surgically excised and, on the basis of histology and immunohistochemistry, diagnosed as a benign teratoma. Six months later, a second histologically similar neoplasm was excised from the same location and was considered to represent local tumor recurrence. The skunk died 16 mo later without recurrence. Complete excision of the tumor appeared curative in this case and supported the benign diagnosis. An extragonadal teratoma should be considered as a differential diagnosis for an abdominal mass in a skunk. PMID- 15526902 TI - Myasthenia gravis in a polar bear (Ursus maritimus). AB - A 14.6-yr-old, female, multiparous polar bear (Ursus maritimus) acutely developed an apparent hind limb weakness. Physical examination and diagnostic tests including a hemogram, serum biochemistry, electrolytes, radiographs, and myelogram did not provide a definitive diagnosis. No improvement in condition was noted during 4 days of supportive care, and the bear was euthanized. An ovoid mass was present in the anterior mediastinum, and a thymoma was confirmed histologically. Compared with control polar bears, elevated serum acetylcholine receptor (AChR) antibodies (0.13 +/- 0.06 nmol/L vs. 0.86 nmol/L) were detected by immunoprecipitation radioimmunoassay, which is consistent with myasthenia gravis (MG) in other species. Although the AChR antibody test has not been validated in the polar bear, we are confident in the postmortem diagnosis of MG, which is commonly associated with thymoma in other species. PMID- 15526903 TI - Clinical challenge. Marked distention of the coelomic cavity, with homogeneous soft tissue opacity consistent with fluid, and cranial displacement and compression of the lungs and right coxofemoral luxation. PMID- 15526904 TI - Amelioration of cyclosporine nephrotoxicity by irbesartan, A selective AT1 receptor antagonist. AB - Cyclosporine A (CsA), a fungal undecapeptide, is the most common immunosuppressive drug used in organ transplantation and autoimmune diseases. However, nephrotoxicity is the major adverse effect of CsA use. The molecular mechanisms of CsA nephrotoxicity are not well characterized, but more recent studies suggest an involvement of angiotensin II (ANG II) and reactive oxygen species in the development of cyclosporine nephrotoxicity. Induction of heat shock proteins (HSPs) is one of the best-described cellular responses to heat stress, hypoxia, and exposure to oxidants. HSPs have beneficial roles in protein processing and protection against cell injury. There is emerging evidence that ANG II induces oxidative stress in vitro and in vivo. This study was thus designed to investigate the role of Angiotensin II type I (AT1) receptor antagonist, irbesartan, on CsA-induced nephrotoxicity. Five groups of rats were employed in this study: group 1 served as control, group 2 rats were treated with CsA (20 mg kg(-1), subcutaneously for 21 days), and groups 3, 4, and 5 received CsA along with irbesartan (10, 25, and 50 mg kg(-1), perorally 24 hr before and 21 days concurrently), respectively. Renal function was assessed by measuring serum creatinine, blood urea nitrogen, creatinine, and urea clearance. The renal oxidative stress was measured by renal malondialdehyde levels, reduced glutathione levels, and enzymatic activity of catalase, glutathione reductase, and superoxide dismutase. Renal morphological alterations were assessed by histopathological examination. CsA administration for 21 days resulted in a marked renal oxidative stress and significantly deranged the renal functions as well as renal morphology. All these factors were significantly improved by irbesartan (50 mg kg(-1)) treatment. HSP72, HSP47, and HSP25 were clearly induced and expressed in CsA-treated animals. The induction and expression of HSP25 was markedly protected by treatment with irbesartan, whereas the induction and expression of HSP47 and HSP72 remained unaltered with the irbesartan treatment. These results clearly demonstrate the pivotal role of ANG II-induced oxidative stress and therapeutic potential of AT, receptor antagonist in ameliorating CsA induced nephrotoxicity. PMID- 15526905 TI - Effects of exogenous melatonin on myoglobinuric acute renal failure in the rats. AB - Free oxygen radicals and nitric oxide (NO) play a crucial role in the pathogenesis of myoglobinuric acute renal failure (ARF). In this study, we aimed to investigate the effect of melatonin, a potent free radical scavenger, on the myoglobinuric ARF formed by injecting hypertonic glycerol intramuscularly (i.m.). The rats were randomly divided into 4 Groups. Rats in Group 1 were given saline and those in Groups 2, 3, and 4 were injected with glycerol (10 mL/kg) i.m. Concomitant and 24 hours after glycerol injection Group 3 (5 mg/kg) and Group 4 (10 mg/kg) were administrated melatonin intraperitoneally. Forty-eight hours after the glycerol injection, the blood and kidneys of the rats were taken under anesthesia. Kidney morphology and the levels of urea, creatinine and nitric oxide metabolites (NOx) in the plasma and the enzyme activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and the level of malondialdehyde (MDA) in the kidney were determined. In both groups of melatonin administration, there was no protective effect of melatonin. Moreover, melatonin significantly decreased the level of NO. As a result, we suggest that the decreasing effect of melatonin on NO, which is a strong vasodilatator, may further increase the renal ischemia in this model. Thus, melatonin may have worsening rather than beneficial effects on myoglobinuric ARF. PMID- 15526906 TI - Does the analysis based on a histological and immunohistochemical grading system in the model of BDL kidney allow the quantification of the degree of injury? AB - The aim of this study is to evaluate histopathological findings induced by Nomega nitro-L-arginine methyl ester (L-NAME) and molsidomine (MOL) on the kidney of bile duct ligated rats. Forty Sprague-Dawley rats, each weighing 125 to 140 g, were included in the study. Extent of histological glomerular injury scores (GIS), arterial injury scores (AIS), and tubulointerstitial injury scores (TIS) in each animal were graded. Alpha-smooth muscle actin (alpha-SMA), tenascin, lectin (Ulex europaeus agglutinin-1), and vimentin were used to determine extent of the injury. The cholestasis was evidenced by a significant increase in the levels of serum total bilirubin in BDL rats (p < 0.01). Malondialdeyde MDA levels increased by the bile duct ligation (BDL) to 12.10 +/- 0.45. This value was significantly higher than the other groups (p < 0.01). Changes in the BDL kidney were marked at 7 days after surgery. GIS were observed to have the highest score, especially at juxtamedullary region in BDL/L-NAME rats, and AIS were also the highest score in this region. These observations were lower in BDL/MOL rats. There is a correlation between GIS and AIS scores (r = .2, p < .01). TIS revealed that BDL/L-NAME rats were significantly more damage than rats in the other groups (p<.001). MOL-treated rats showed considerably fewer lesions in the tubules and interstitium (p < .001). The tubular injuries observed in BDL and BDL/L-NAME rats were significantly attenuated by MOL treatment. Lectin was more and extensively stained in tubular epithelia of the BDL/L-NAME group than in the other (p <.05). Expression of tenascin in tubular epithelia was significantly higher in BDL and BDL/L-NAME as compared with controls (p < .01). Fibrous tissue was only observed in the BDL and BDL/L-NAME group. These areas were weakly stained with vimentin. alpha-SMA staining was more reduced in the L-NAME-treated arterioles than in BDL/MOL (p < .05). In conclusion, the analysis of cell injury based on a histological grading system in the model of BDL kidney allows the quantification of the degree of injury. PMID- 15526907 TI - Anti-inflammatory effect of PPARgamma in cultured human mesangial cells. AB - Our aim is to investigate whether peroxisome proliferator-activator receptor gamma (PPARgamma) expression was altered in human mesangial cells under inflammatory stress and whether PPARgamma could retard the inflammatory responses. Based on cultured human mesangial cell lines (HMCLs), PPARgamma expressions at protein and mRNA levels were observed by Western blot analysis and reverse transcriptase polymerase chain reaction. Informatory cytokines such as tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Our results demonstrated that PPARgamma protein expression was dramatically increased in HMCLs stimulated by IL-1beta (10 ng/mL). The levels of IL-6 and TNF-alpha in HMCL supernatants, protein, and mRNA expressions of PPARgamma in IL-1beta challenge cells were significantly increased more than those in untreated cells. Importantly, PPARgamma agonists troglitazone, rosiglitazone, and 15-deoxy-delta(12, 14)-prosglandin J2 significantly decreased the up expression of TNF-alpha and IL-6 in HMCL supernatants stimulated by IL 1beta. Furthermore, troglitazone downregulated TNF-alpha and IL-6 mRNA expression from IL-1beta challenge HMCLs. Our data suggest that PPARgamma plays an important role in mesangial cells responding to inflammatory stress. PPARgamma may prove to be a pharmacological target in glomerulonephritis. PMID- 15526908 TI - Are bile acids involved in the renal dysfunction of obstructive jaundice? An experimental study in bile duct ligated rats. AB - BACKGROUND: Surgery on patients with obstructive jaundice is associated with a significant risk of postoperative renal failure. Bile acids are implicated as nephrotoxins because they accumulate in the plasma and the kidney becomes their only excretory route in cholestasis. The experimental evidence favoring this proposal is inadequate and unconvincing. Therefore, we designed an animal experiment involving bile duct ligated (BDL) rats in which we could correlate variations in serum and urine bile acids with indices of nephrotoxicity and renal function. HYPOTHESIS: Bile acids are putative nephrotoxins. MATERIALS AND METHODS: Total serum and urine bile acid concentrations and profiles were determined using liquid chromatography/gas chromatography/mass spectrometry selected ion monitoring. Nephrotoxicity was assessed by renal histopathology and by determination of the urinary activities of the following enzymes: muramidase, glutamate dehydrogenase, alkaline phosphatase, N-acetyl-beta-D-glucosaminidase, and lactate dehydrogenase. Renal function was assessed by measuring urine osmolality, daily osmolar excretion, sodium excretion (U(Na)V), potassium excretion (U(K)V), and total protein and albumin excretion. RESULTS: Maximum plasma concentrations and renal clearance of bile acids occurred between the third or fourth postoperative day following BDL. This peak coincided with maximal disruption of proximal convoluted tubule architecture and postoperative changes in renal function-increased urine flow rate and decreases in urine osmolality and sodium excretion. Thereafter, 1) plasma levels of bile acids returned toward normal levels, 2) urinary bile acid clearance declined, 3) normal renal histology was restored, and 4) normal renal function was reestablished. Throughout this period, fluctuations in enzymuria were evident. However, these shifts did not coincide with plasma and urine bile acid concentrations and histological and functional changes. DISCUSSION AND CONCLUSIONS: Transient functional impairment of renal cation and water transport and nonspecific morphological changes in the proximal convoluted tubule occur 3 to 4 days following bile duct ligation in rats. These functional and morphological changes occurred when plasma total and urinary bile acids were at their peaks. Although it is tempting to equate association with causality, we cannot implicate bile acids as being responsible for the aberrations in renal function and structure following BDL. Accordingly, we have concluded that elevated plasma concentrations of bile acids are renal exacerbates acting in concert with other factors, be they prerenal or renal in origin to precipitate a cascade of events leading to postoperative renal failure in cholestasis. PMID- 15526910 TI - Renal artery stenosis of solitary kidney: the dilemma. AB - BACKGROUND: Renal artery stenosis is an important cause of hypertension and renal failure. When present in a solitary kidney, it is associated with extremely high morbidity and mortality. Although guidelines suggest an aggressive approach by revascularization in these cases, available evidence is too weak and contradictory to provide definite recommendations for treatment. OBJECTIVES: To discuss the various clinical factors predicting the benefit from revascularization. METHODS AND RESULTS: Review of more recent clinical trials is presented. DISCUSSION: The approach to the patient with single kidney and renal artery stenosis is discussed and, although there is insufficient evidence-based medicine for guidelines, some suggestions are made to predict revascularization results in these cases. CONCLUSION: Continuous development of clinical prediction rules will help in the management of renal artery stenosis of solitary kidney. PMID- 15526909 TI - Attenuation of chloroquine-induced renal damage by alpha-lipoic acid: possible antioxidant mechanism. AB - The toxic effect of chloroquine (CQ) has been attributed to oxidative stress with the consequences of lipid peroxidation. This study investigates the effects of alpha-lipoic acid (LA) on CQ-induced nephrotoxicity in rats. A single oral administration of CQ (970 mg/kg)-induced nephrotoxicity, manifested biochemically by a significant increase in serum creatinine and blood urea nitrogen concentrations. In addition, renal tissue from CQ-treated rats showed a significant increase in lipid peroxides measured as thiobarbituric acid reactive substances and hydroperoxides, along with significant decrease in nonenzymic antioxidants (vitamin C, vitamin E, and reduced glutathione) and enzymic antioxidants (superoxide dismutase, catalase, glutathione peroxidase, and glutathione-S-transferase) levels. Oral administration of LA (10, 30, or 100 mg/kg) in different doses for 10 days produced a significant protection against nephrotoxicity induced by CQ. Treatment with LA markedly reduced the elevated lipid peroxidation, restored the depleted renal antioxidant defense system. LA at 100 mg/kg was effective when compared with other doses (10 and 30 mg/kg). This was accompanied by the histopathological observations in kidney tissue. The results suggest that LA ameliorate the lipid peroxidation and the loss of cellular antioxidants, thereby protecting the CQ-induced oxidative damage in kidney. PMID- 15526911 TI - Early referral as an independent predictor of clinical outcome in end-stage renal disease on hemodialysis and continuous ambulatory peritoneal dialysis. AB - OBJECTIVES: To evaluate the influence of early nephrology referral on clinical outcome in patients on maintenance hemodialysis (HD) and peritoneal dialysis (PD). PATIENTS AND METHODS: This study retrospectively analyzed patients entering our HD and PD program from February 2000 to June 2003. Patients who presented to a nephrologist more than 6 months before starting dialysis were defined as early referral (ER). Meanwhile, patients transferred to the nephrology department less than 6 months before initial dialysis were considered late referral (LR). RESULTS HD GROUPS: Of 78 HD patients, 37 (47.1%) qualified for the ER group and 41 (52.6%) were designated to the LR group. The demographic data were analyzed for both the HD and PD groups. No significant differences in average age at dialysis, duration of hemodialysis, and gender were noted between these two groups. The same applied for the biochemical parameters in both groups. HD patients with early referral had significantly better survival (p < .05) as plotted with the Kaplan-Meier method. In univariate analysis by cox proportional hazards mode, the early referral in HD patients [Exp (Coef) = 0.426, P < .01] significantly influenced survival. The various variables were further examined by multivariate analysis, and early referral, hemoglobin, and age still significantly impacted patient survival (P < .05). CAPD GROUPS: The survival curve related to early referral in continuous ambulatory peritoneal dialysis (CADP) patient survival rate was significantly higher for the early referral groups (P < .05). In addition, a multivariate analysis adjusting for several potential risk factors found that referral time remained significantly associated with patient survival. In additional, hemoglobin and age were significant and independent predictors of mortality. CONCLUSION: This study demonstrates that time between referral and starting dialysis is a predictor of survival for both HD and PD patients, with early referral being associated with longer survival time. These analytical results suggest that early referral before dialysis is important in determining long-term prognosis in HD and PD patients. PMID- 15526912 TI - Effect of exercise training on interdialytic ambulatory and treatment-related blood pressure in hemodialysis patients. AB - BACKGROUND: Exercise training improves blood pressure (BP) in the general population, but prior studies in hemodialysis (HD) patients only used pill counts or treatment-related BPs. We evaluated the effect of 3 to 6 months of intradialytic exercise training on ambulatory blood pressure (ABP) and treatment related pre- and postdialysis BP. PATIENTS AND METHODS: Nineteen chronic HD patients trained with an exercise bicycle for 30 to 60 min in the first 1 to 2 hr of each of thrice weekly HD. Interdialytic 44-hr ABP was performed a week before training began and repeated at 3 and 6 months. Pre- and post-HD systolic and diastolic BP and pre- and post-HD weight were recorded for 2 months prior to training, throughout the training, and, if available, for the 2 months after training ended. BP medications were recorded throughout. Body composition by bioimpedance, and norepinephrine and epinephrine levels by RIA were done at 0, 3, and 6 months. RESULTS: Thirteen subjects who completed at least 3 months of training exercised 90% of HD sessions for 56 min +/- 23 SD each. Systolic and diastolic 44-hr interdialytic ABP fell during training (systolic 138.4 mmHg +/- 19.6 vs. 125.7 mmHg +/- 20.0 vs. 125.9 mmHg +/- 22.9; diastolic 83.2 mmHg +/- 10.2 vs. 74.7 mmHg +/- 9.0 vs. 73.9 mmHg +/- 11.8 at 0, 3, and 6 months; p < .05 ANOVA). Norepinephrine and epinephrine levels did not independently predict systolic BP. Pre-HD systolic BP was stable during the pretraining period, fell significantly during the training period (p < .03), and returned toward preexercise levels during the posttraining period (p < .001). Pre- or postweight, erythropoietin dose, total body water, and number of BP meds were unchanged. CONCLUSION: Exercise training during HD significantly improves both interdialytic ABP and treatment-related BP. PMID- 15526913 TI - Evaluation of prognostic indexes in critical acute renal failure patients. AB - OBJECTIVE: To study different prognostic indexes in acute renal failure (ARF) patients admitted to an intensive care unit (ICU). DESIGN: Prospective, cohort study. Individual Severity Score-Acute Tubular Necrosis (ISS-ATN) obtained prospectively and retrospectively, Acute Physiologic and Chronic Health Evaluation (APACHE II) Score, APACHE II Risk, Lung Injury Score (LIS), and Number of Organ Failures (NOF) were calculated for each patient. The outcome analyzed was death in the ICU. Discrimination was evaluated by the area under the receiver operator characteristic curve (AUC). For calibration analysis, the chi-square goodness-of-fit test was used to compare predicted mortality, calculated by ISS ATN (obtained prospectively or retrospectively) and APACHE II risk, with observed mortality. SETTING: ICU, Sao Rafael Hospital, Salvador-BA, Brazil. PATIENTS: Seventy-six ARF patients admitted to the ICU within 6 months. INTERVENTIONS: Surgical and medical procedures. MEASUREMENTS AND RESULTS: The observed AUC was 0.69 for LIS, 0.73 for prospective ISS-ATN, 0.75 for retrospective ISS-ATN, 0.76 for APACHE II Score, 0.78 for APACHE II Risk, and 0.88 for NOF. These areas were significantly different from 0.5 (p < .001). There was no difference between the observed and expected death rate calculated by ISS-ATN. However, APACHE II underestimated the observed mortality (p < .001). CONCLUSION: The prognostic indexes studied showed good discriminative power. However, APACHE II was not well calibrated in contrast to the good calibration of ISS-ATN. PMID- 15526914 TI - Anti-N-like and anti-Form red cell antibodies in chronic hemodialysis patients. AB - Allogeneic red blood cell (RBC) transfusions and the use of reusable dialyzers sterilized with formaldehyde can lead to RBC alloimmunization in chronic hemodialysis patients. The formed RBC alloantibodies have been implicated in immediate kidney allograft failure and decreased RBC survival observed in these patients. Using indirect antiglobulin test, direct antiglobulin test (DAT), and direct Polibrene test (DPT), we detected an RBC alloimmunization rate of 17.2% (11/64) in transfused hemodialysis patients, and found the presence of anti-N like and anti-Form antibodies in 5 (5.7%) and 53 (60.9%) individuals, respectively. The sensitivity rate of the DPT was significantly higher than that of the DAT in detecting anti-Form, but the DAT showed a higher specificity rate compared with the DPT. We conclude that patients treated with reusable dialyzers sterilized with formaldehyde may develop specific RBC alloantibodies that could increase the potential risk of hemolysis, decrease survival of RBCs, and increase the need of blood supply. PMID- 15526915 TI - Maternal and fetal outcomes in HELLP syndrome complicated with acute renal failure. AB - OBJECTIVE: This study reviews maternal and fetal outcomes in HELLP syndrome complicated with acute renal failure (ARF), and compares clinical and laboratory findings of the cases of HELLP syndrome that did not develop ARF. MATERIALS AND METHODS: All pregnant women with hypertensive disorders admitted or referred to the maternal and fetal unit were recorded into a perinatal database between January 15, 2002 and September 15, 2003. During the study period, out of 615 cases of hypertensive pregnancy, we followed and delivered 347 cases of severe preeclampsia, of them 132 cases were diagnosed as HELLP syndrome. ARF was defined as creatinine level > or =1.2 mg/dL and/or oliguria <400 mL/24 hr. The cases were divided into three groups on the basis of the highest creatinine level recorded during hospitalization: creatinine <1.2 mg/dL, creatinine > or =1.2 to 2.0 mg/dL, and creatinine > or =2.0 mg/dL. Statistical comparisons were performed by Student t test, X2 analysis, and Fisher's Exact test as appropriate. The value of P < .05 was considered significant. RESULTS: ARF developed in 8.9% (n:31) of severe preeclampsia (n:347); of them, 15 (4.3%) cases were nonoliguric, and all had mildly elevated creatinine levels between 1.2 and 1.9 mg/dL. Moderately elevated creatinine levels were 2 to 3.9 mg/dL in 10 cases, and severely elevated creatinine levels were 4 to 8.4 mg/dL in 6 cases, for a total of 16 (4.6%) cases; creatinine levels were > or =2.0 mg/dL (range: 2.0-8.4 mg/dL). HELLP syndrome was the most frequent cause of ARF, 64.5% (n:20/31), and was observed in 15% (n:20) of 132 cases of HELLP syndrome. Fourteen (88%) of 16 cases that had oliguria and creatinine levels > or =2 mg/dL were detected in HELLP syndrome (n:14/132; 10.6%). Major maternal complications in HELLP syndrome with ARF and creatinine level > or =2 mg/dL in the study group were abruptio placentae (42.8%; n:6/14), incisional hematoma (21%; n:3/14), pulmonary edema (14%; n:2/14), cesarean hysterectomy (7%; n: 1/14), and dialysis (50%; n:7/14). There was no maternal mortality. All patients complicated with ARF were discharged without renal impairment. Perinatal mortality was 26.1% in the cases of HELLP syndrome with ARF creatinine > or =1.2 mg/dL and further increased to 37.5% when creatinine levels were above 2.0 mg/dL, compared with 11.8% in the cases having creatinine <2.0 mg/dL, and the difference was statistically significant (p:.007). CONCLUSIONS: The most contributing factors leading to ARF in HELLP syndrome were abruptio placentae and HELLP syndrome complicated with ARF, particularly, oliguric ARF has relatively higher maternal complications and perinatal mortality. PMID- 15526916 TI - The clinical course of patients with type 1 hepatorenal syndrome maintained on hemodialysis. AB - GOAL: Report the natural coarse of hepatorenal syndrome in 4 patients who were maintained on chronic hemodialysis. BACKGROUND: The diagnosis of hepatorenal syndrome carries a grave prognosis with a mortality rate >90% and a median survival time of <2 weeks without orthotopic liver transplantation. STUDY: We report the clinical course of 4 patients with hepatorenal syndrome who underwent long-term (greater than 3 weeks) hemodialysis in an attempt to bridge them to orthotopic liver transplantation. The etiologies of cirrhosis were: chronic hepatitis C infection (n = 2), alcoholic liver disease (n = 1), and primary sclerosing cholangitis (n = 1). RESULTS: Mean survival time on hemodialysis was 236 days (range: 31 to 460 days). All patients survived their initial hospitalization and were discharged from the hospital. However, only one patient received orthotopic liver transplantation. Mean number of hospital admissions was 11 (range: 4 to 18) while receiving hemodialysis at an average rate of 2.2 (range: 1.1 to 5) admissions/patient month. Mean number of days spent in hospital while on hemodialysis support was 85 days (range: 15 to 199 days) at an average rate of 11.2 (range: 8.3 to 15) hospital days/patient month. An average of 33% (range: 26% to 48%) of the days of the prolonged survival on hemodialysis was spent in hospital. CONCLUSION: Although our 4 patients with hepatorenal syndrome demonstrated long-term survival with hemodialysis, their prolonged survival was at the cost of a very heavy burden of morbidity and in-patient stay. The advisability of maintenance hemodialysis in patients with hepatorenal syndrome should be judged on an individual basis. PMID- 15526917 TI - Therapeutic apheresis of immune diseases in nephrology department. AB - OBJECTIVE: The clinical efficacy of therapeutic apheresis is still controversial. We undertook a retrospective review of apheresis treatment to ascertain its safety and efficacy. METHODS: We reviewed 31 patients (13 male, 18 female). Plasmapheresis was performed on 7 patients with hematologic disorders, 5 patients with neurologic disorders, 6 patients with systemic diseases, and 3 patients with Lyell syndrome. Immunoadsorption onto protein A sepharose was evaluated as rescue therapy in 7 patients. Low-density lipoprotein (LDL) apheresis was performed on 3 patients. RESULTS: There were five mortalities due to serious complications of their primary disease. Most complications were mild such as hypotension and hypocalcemia. Two patients who received LDL apheresis had severe anaphylactic reactions. Apheresis was effective in the remaining 24 patients. CONCLUSIONS: The therapeutic apheresis consists of a continuously improving therapeutic method for diseases with high mortality and morbidity, especially in cases with poor outcome by using current medications. PMID- 15526918 TI - Cognitive status in hemodialysis as a function of fluid adherence. AB - Previous work has shown that dialysis improves cognitive functioning in hemodialysis patients, perhaps due to improvements in anemia among these patients. Such improvements in cognitive performance may lead to better levels of self-care and adherence with treatment. This study examined the relationship between fluid adherence and cognitive functioning in patients receiving hemodialysis. One hundred forty-seven patients were assessed with a brief screening instrument, the Cognistat, to determine their current level of functioning during the first hour of hemodialysis. Fluid nonadherence was operationalized as interdialytic weight gain above 1 kg/day. Rates of impairment on the Cognistat subscale ranged from 2.7% (orientation) to 54% (memory) in this sample. Roughly 68% of the sample was nonadherent during the course of treatment. Results found no differences in mean levels of cognitive performance between those who were adherent and those who were not and only modest relationships of measures of anemia to certain aspects of cognitive performance. For the hemodialysis patient to benefit from self-care education, the patient must be able to understand, remember, reason, and use cognitive processes to modify behavior. These results suggest that more in-depth assessment of cognitive performance may be needed. In addition, this assessment may need to be conducted on a day when treatment is not received. PMID- 15526919 TI - A six-year follow-up after interferon-alpha monotherapy for chronic hepatitis C infection in hemodialysis patients. AB - Interferon-alpha (IFN) has been accepted as an effective treatment for chronic hepatitis C virus (HCV) infection in hemodialysis (HD) patients. We prospectively assess the long-term clinical, biochemical, and virological effects of interferon in the treatment of HD patients with chronic HCV infection. This study was performed in 20 HCV-RNA-positive HD patients with evidence of chronic hepatitis on liver biopsy. The patients received IFN administered after HD sessions in doses ranging from 3 to 6 million units for 6 to 12 months. The patients were followed up for a period of 6 years with determinations of serial alanine aminotransferase (ALT) levels and serum HCV-RNA. At the time of the final follow up, the patients had no cirrhosis or hepatocellular carcinoma. Among the nonresponder group, only 1 patient died due to sudden cardiac death. Sustained normal serum ALT levels occurred in 9 (45%) of the patients. Nine patients had variable ALT levels, and 2 patients had persistently elevated ALT levels. Eight (40%) patients were continuously HCV-RNA negative, whereas 12 patients (60%) had variable HCV-RNA results at the end of the 6-year follow-up. These findings show that the long-term clinical, biochemical, and virological response to interferon monotherapy is good in HD patients with HCV infection. PMID- 15526920 TI - Urinary excretion of vitamin A in critically ill patients complicated with acute renal failure. AB - OBJECTIVE: Study the possible excretion of vitamin A in urine of critically ill patients complicated with acute renal failure. PATIENTS AND METHODS: Nine Intensive Care Unit patients, age 71.2 +/- 15.7 (mean +/- SD) with acute renal failure were studied. Urinary retinol, creatinine, protein, albumin, and serum creatinine were measured. RESULTS: All patients excreted retinol in urine; individual values ranged from 0.007 to 0.379 micromol retinol/mmol creatinine. There was no correlation of urinary retinol/creatinine ratio with serum creatinine or with urinary protein/creatinine and albumin/creatinine ratios. CONCLUSION: Excretion of retinol in urine may be indicative of acute renal failure in critically ill patients. PMID- 15526921 TI - Waldenstrom's macroglobulinemia presenting as Goodpasture's syndrome. AB - We report a case of Waldenstrom's macroglobulinemia (WM) presenting as classic Goodpasture's syndrome. Thrombocytopenia, an additional autoimmune phenomenon, further complicated the clinical course. Renal disease is well recognized in Waldenstrom's macroglobulinemia and other B-cell dyscrasias. Similarly, a wide spectrum of autoimmune-mediated conditions has been reported in association with paraproteinemia. However, rapidly progressive glomerulonephritis in association with antiglomerular basement membrane antibodies has never been reported as either an autoimmune or a renal manifestation of WM. This article reviews the current understanding of renal pathology and autoimmune phenomenon associated with WM. PMID- 15526922 TI - Circumpolar health research. PMID- 15526923 TI - Public health research and practice in Greenland. PMID- 15526924 TI - A leading authority on cold injuries in Alaska. PMID- 15526925 TI - Impact of public health research in Greenland. AB - In 1992, the Greenland Home Rule Government took over the responsibility for health care. There has since been a growing cooperation between the Directorate of Health and researchers in Denmark and Greenland, for instance by the Directorate supporting workshops and funding a chair in health research. Two health surveys have been carried out in Greenland by the National Institute of Public Health, and a follow-up is being planned together with the Directorate of Health. The results have been widely used by politicians, administrators, and health care professionals. PMID- 15526926 TI - Mental health of Sami youth. AB - Sami children and adolescents are the indigenous youngsters in Norway, mainly resided in the arctic part of the country. While disadvantaged living conditions, risk behavior and psychososial health problems has been shown for children and youth from many indigenous groups worldwide, the research among Sami youngsters is sparse. However, recent research show that compared to the Norwegian majority group, Sami children and adolescents have just as good mental health as their majority peers. They also show less risk taking behavior as substance and drug use, have less eating problems and have a stronger body satisfaction. However, the smoking rates are high as for their Norwegian counterparts. Intragroup studies show that Sami adolescents grown up in Sami dominated areas, have a strong bicultural identification, are practicing more Sami cultural behavior and have a better mental health compared to Sami peers in marginal Sami areas. Ethnocultural factors have only a slight impact on behavior problems among young Sami and particularly among boys in the marginal Sami areas. PMID- 15526927 TI - Impact of economic changes on the diet of Chukotka Natives. AB - This paper describes and analyses changes in food composition and nutritional preferences among the Chukchi and Yupik of coastal Chukotka in the last 15 years. The economic collapse of the infrastructure of Chukotka region has resulted in many indigenous northerners reverting to the traditional subsistence economy. Relatively expensive market foods are being replaced by cheaper ones, and by more readily available local foods. Percent contribution of proteins, lipids and carbohydrates to total caloric intake has not changed substantially, but sources of the major nutrients have become different. In 1985, local marine mammals accounted for about half of the consumed meat (55%), while in 2000 the share of it increased to 89 %. Market fats and oils are also being substituted by the fat of marine mammals. However, the contemporary diet of the natives of coastal Chukotka differs significantly from the traditional one. The meat of seals and gray whales (small sized and less dangerous to harvest) remains seasonally accessible, but can not be stored for long times. There is an insufficient amount of walrus and bowhead whale meat, which can be prepared in traditional style by fermentation, and stored for a long time. This probably also provides a specific protection against Helicobacter pylori. The young people today are more oriented towards local food-stuff: 76 % Coastal Chukchee and Yupik under the age of 30 indicated a preference for native foods over European ("Russian") ones, while this share is lower (66 %) among people older than 30 years. Overall, 86 % of natives consider that whale hunting, as the main source of food, should be increased (in 1985, only 45% suggested so). PMID- 15526928 TI - Healthy living in Nunavut: an on-line nutrition course for inuit communities in the Canadian arctic. AB - OBJECTIVES: It is recognized that empowerment of Indigenous Peoples through training and education is a priority. The objective was to design a course that would provide an innovative training approach to targeted workers in remote communities and enhance learning related to the Nunavut Food Guide, traditional food and nutrition, and diabetes prevention. STUDY DESIGN: A steering committee was established at the outset of the project with representation from McGill University and the Government of Nunavut (including nutritionists, community nurses and community health representatives (CHRs), as well as with members of the target audience. Course content and implementation, as well as recruitment of the target audience, were carried out with guidance from the steering committee. METHODS: An 8-week long course was developed for delivery in January - March, 2004. Learning activities included presentation of the course content through stories, online self-assessment quizzes, time-independent online discussions and telephone-based discussions. Invitations were extended to all prenatal nutrition program workers, CHRs, CHR students, home-care workers, Aboriginal Diabetes Initiative workers and public health nurses in Nunavut. RESULTS: Ninety-six health-care workers registered for Healthy Living in Nunavut, with 44 actively participating, 23 with less active participation and 29 who did not participate. CONCLUSIONS: Despite having to overcome numerous technological, linguistic and cultural barriers, approximately 40% of registrants actively participated in the online nutrition course. The internet may be a useful medium for delivery of information to target audiences in the North. PMID- 15526929 TI - Heart defects and other malformations in the Inuit in Canada: a baseline study. AB - OBJECTIVES: Birth defects occur in all ethnic groups, remaining an important world-wide cause of perinatal and infant morbidity. This contributes greatly to an excess of health care dollars allocated to the care and repair of those affected. This is especially true when those affected live in remote geographical locations. STUDY DESIGN: A chart review of 2567 live births of children of Inuit parents residing in Arctic Quebec (Nunavik) and on Baffin Island (Nunavut) between 1989 and 1994 (five years) was carried out compared to rates of anomalies of the Alberta Congenital Anomalies Surveillance System (ACASS). RESULTS: Birth defects were higher in the Inuit sample in nearly every major ICD-9 category with the exception of neural tube defects, eye anomalies and chromosome abnormalities. (Total: 99.7/1000 Vs 51.5/1000; OR 1.93 95% CI 1.7-2.3). Congenital heart defects were significantly increased 22.9/1000 Vs 5.6/1000, with an OR of 4.18 (95% CI 3.2-5.4) in the ICD-9 category 745. In particular, ventricular septal defects (VSDs) and atrial septal defects (ASDs) (OR 4.9 CI 3.5-6.9 and 4.6 CI 2.9-7.2) were frequent. CONCLUSIONS: A high rate of heart defects was an important contributor to the nearly two times rate of total birth defects in the Inuit compared to the ACASS. Further study should be carried out to determine the contributing factors. Genetic predisposition to specific heart defects, and a diet low in folate and vitamin A are considerations. The use of alcohol may exacerbate vitamin status in pregnancy. Optimizing vitamin status in the periconceptional period may reduce the rate of birth defects. PMID- 15526930 TI - Daylight availability: a poor predictor of depression in Iceland. AB - OBJECTIVES: To test the hypothesis that the unexpectedly low prevalence of winter depression in Iceland is explained by Icelanders enjoying more daylight, during the winter months, than allocated to them by latitude. METHODS: A conventional photometer was applied to measure illuminance on a horizontal surface at 64 degrees 8.8' N and 21 degrees 55.8' W every minute throughout the year. The illuminance thus measured was compared with computed illuminance, based on theoretical upper bounds. RESULTS: Daylight availability proved to be, on average, 60% of the theoretical upper bounds derived using clear sky conditions. Snow cover did not, on average, cause a significant increase in daylight availability. Great variability was observed in illuminance from day to day, as well as within days. CONCLUSIONS: Average daylight availability does not explain the lower than expected prevalence of winter depression in Iceland. The great variability in illuminance might, however, affect the expression of winter depression, as could daylight quality and genetic factors. PMID- 15526931 TI - The current status of diabetes care, diabetic retinopathy screening and eye-care in British Columbia's First Nations Communities. AB - OBJECTIVES: The objective of this study was to evaluate the current status of primary diabetes care, diabetic retinopathy screening, and eye-care for First Nations individuals with diabetes living on-reserve in British Columbia. STUDY DESIGN: Prospective cross-sectional observational survey. METHODS: A mail-out survey was sent to all BC First Nations Communities in the format of a 26-item questionnaire. A series of general questions were asked concerning community populations, regional transportation options and the availability of local health care providers. Specific questions about the frequency and source of eye and diabetic retinopathy care in each community were also solicited. RESULTS: A sixty seven percent questionnaire response rate was achieved (136/202 communities). The community-reported rate of diabetes mellitus ranged from 4.8 to 11.8% with an average of 6.4%. The proportion of on-reserve individuals with diabetes receiving yearly retinal examinations for diabetic retinopathy was found to be only 33%. Many communities received eye-care from more than one professional group, but the majority of basic eye-care and retinopathy screening was provided by optometrists. Ophthalmologists were less likely to provide eye care for these communities; however, family physicians and nurses were frequently responsible for performing eye evaluations. CONCLUSIONS: The vast majority of First Nations people with diabetes who live on-reserve in British Columbia do not have access to annual examinations by an eye-care professional. Eye evaluations and retinal screening were often the responsibility of individuals with little formal training in this area. PMID- 15526932 TI - Glucose intolerance and its relation to cardiovascular risk factors among Greenland Inuit. The Greenland Population Study. PMID- 15526933 TI - The teleconsultation in general practice. A randomized, controlled study of a remote consultation experiment using a videoconferencing system. PMID- 15526934 TI - Making gene-modified mice. PMID- 15526935 TI - High-throughput gene knockouts and phenotyping in mice. PMID- 15526936 TI - Modelling gene-environment interactions in Th1- and Th2-dominated diseases of laboratory animals. PMID- 15526937 TI - Animal models of experimental asthma. PMID- 15526938 TI - Models of rheumatoid arthritis. PMID- 15526939 TI - Animal models of inflammatory bowel diseases. PMID- 15526940 TI - Murine models of atopic dermatitis. PMID- 15526941 TI - Epicutaneous sensitization with allergens as an atopic dermatitis model. PMID- 15526942 TI - T-cell receptor transgenic models of inflammatory disorders: relevance for atopic dermatitis? PMID- 15526943 TI - What must a model display for proof as a model of psoriasis? PMID- 15526944 TI - From classical mouse models of psoriasis to a spontaneous xenograft model featuring use of AGR mice. PMID- 15526945 TI - The psoriasis SCID mouse model: a tool for drug discovery? PMID- 15526946 TI - Pro- and anti-inflammatory effects of IL-4: from studies in mice to therapy of autoimmune diseases in humans. PMID- 15526947 TI - What must a model display for proof as a model of contact dermatitis? PMID- 15526948 TI - Acute and chronic models of allergic contact dermatitis: advantages and limitations. PMID- 15526949 TI - Transgenic mice expressing IFN-gamma in the epidermis are a model of inflammatory skin disease and systemic lupus erythematosus. PMID- 15526950 TI - Neurobiology of specific language impairment. AB - This review summarizes what is known about the neurobiology of specific language impairment. Despite its name, specific language impairment is frequently not specific. It is common to find associated impairments in motor skills, cognitive function, attention, and reading in children who meet criteria for specific language impairment. There is evidence that limitation in phonologic working memory may be a core deficit in specific language impairment. Both genetic and environmental factors have been shown to be important etiologic factors in specific language impairment. Structural neuroimaging studies suggest that atypical patterns of asymmetry of language cortex, white-matter abnormalities, and cortical dysplasia may be associated with specific language impairment. Abnormalities in the later stages of auditory processing have been demonstrated using auditory event-related potentials. Functional neuroimaging may cast further light on the neurobiology of specific language impairment and serve as a means of developing and evaluating therapy. A better understanding of the neurobiology of specific language impairment is critical for the rational development of therapeutic strategies to treat this common disorder. PMID- 15526951 TI - Neurocysticercosis in children. AB - Neurocysticercosis is a major cause of neurologic disease worldwide. The clinical presentations are pleomorphic depending on the stage and location of cysts in the nervous system. Most children (> 80%) present with seizures, particularly partial seizures; headache and vomiting are seen in about a third of cases. Diagnosis is made by either computed tomography (CT) or magnetic resonance imaging. Single enhancing lesions are the most common finding. Visualization of a scolex confirms the diagnosis. Some cases have multiple cysts; the "starry-sky" appearance in cases with innumerable cysts is characteristic. Most children require anticonvulsants. Corticosteroids are indicated in those with cerebral edema. The efficacy of cysticidal therapy continues to be debated. Controlled studies suggest that cysticidal therapy helps in increased and faster resolution of CT lesions, but there is no conclusive evidence that it improves long-term seizure control. The prognosis in cases with single lesions is good; seizure control is achieved with a single anticonvulsant, and the recurrence rate is low. Children with multiple lesions have recurrent seizures. Extraparenchymal neurocysticercosis is rare in children and carries a poor prognosis. Neurocysticercosis must be considered in the differential diagnosis of seizures and a wide variety of neurologic disorders, particularly in endemic areas. PMID- 15526952 TI - Neonatal arterial ischemic stroke and cerebral sinovenous thrombosis are more commonly diagnosed in boys. AB - The risk factors for arterial ischemic stroke and cerebral sinovenous thrombosis in neonates are not well understood. We looked at gender, birthweight, and gestational age in neonates with arterial ischemic stroke and cerebral sinovenous thrombosis to see if there were trends suggesting that these were risk factors. We identified neonates with a gestational age at birth > or = 36 weeks and a diagnosis of arterial ischemic stroke or cerebral sinovenous thrombosis made by computed tomography or magnetic resonance imaging during the neonatal period from a consecutive cohort study of children with arterial ischemic stroke and cerebral sinovenous thrombosis in Ontario. Data on gender, birthweight, and gestational age were obtained by health record review. Sixty-six children with neonatal arterial ischemic stroke were identified. Forty-one (62.1%; 95% CI 49.3-73.8%) were male. Thirty-two children with neonatal cerebral sinovenous thrombosis were identified. Twenty-five (78.1%; 95% CI 60.0-90.7%) were male. One male child was identified with both arterial ischemic stroke and cerebral sinovenous thrombosis. There was a trend toward higher than average birthweights among neonates with arterial ischemic stroke and a trend toward older gestational age in female neonates with arterial ischemic stroke. Our data suggest that neonatal arterial ischemic stroke and cerebral sinovenous thrombosis are more commonly diagnosed in boys. The slightly larger size of male neonates may be contributory in arterial ischemic stroke. It is not known whether boys are at higher risk of developing arterial ischemic stroke and cerebral sinovenous thrombosis or are simply more likely to present with symptoms resulting in diagnosis. These issues need further study. PMID- 15526953 TI - Valproic acid-induced pancreatitis in childhood epilepsy: case series and review. AB - In the past 6 years, 11 children on valproic acid have developed pancreatitis in our children's hospital. Valproic acid has been used as one of the primary anticonvulsants for generalized seizures in children for the past 25 years. A literature review reveals mostly singular reports of pancreatitis over the past decade. The charts of the 11 patients with valproic acid-induced pancreatitis were reviewed. Dosage, valproic acid serum levels, duration of therapy, and concomitant medications were examined. Families were contacted by telephone to determine the formulation (brand name vs generic) of valproic acid at the time of diagnosis. Six girls and five boys were studied. The ages ranged from 4 to 16 years. Eight of 11 children presented with an acute abdomen. Unexpectedly, three children presented with a flulike illness. Serum lipase values ranged from 341 to 5576 U/L (normal range < 190 U/L). The dose of valproic acid ranged from 20 to 50 mg/kg. Serum levels ranged from 334 to 884 micromol/L (therapeutic range 350-800 micromol/L). Six of the patients were on monotherapy. Seven children were on brand-name drugs. Four of the children had an abnormal neurologic syndromic diagnosis (West syndrome, Rett syndrome, Lowe syndrome, and Angelman's syndrome). Six of the children had a history of drug allergies with a skin rash. Valproic acid was reintroduced in one child and resulted in a second episode of pancreatitis. Resolution of symptoms usually took several weeks following discontinuation of the drug. No association was found with valproic acid dosage, type of preparation, serum levels, duration of therapy, or presence of concomitant medications. Pancreatitis is a severe adverse effect of valproic acid use in children. Dose, duration of treatment, serum valproic acid levels, generic preparation, and the presence of concomitant antiepileptic drugs do not appear to be risk factors. Children with known drug sensitivity might be at risk. Lipase levels at the time of an acute abdomen or a flulike illness in epileptic children taking valproic acid can reveal early stages of pancreatitis and are recommended. PMID- 15526954 TI - Influence of MECP2 gene mutation and X-chromosome inactivation on the Rett syndrome phenotype. AB - To date, approximately 200 different mutations in the MECP2 gene have been identified. We analyzed the entire coding sequence of the MECP2 gene and the X chromosome inactivation pattern in 42 sporadic cases of Rett syndrome. Of the 42 patients, 30 had pathogenic mutations, including 14 different mutations: 9 missense mutations, 4 nonsense mutations, and 1 frameshift mutation. One was a novel mutation (S134P). There was a tendency for patients who had a nonsense mutation in the transcriptional repression domain region to show earlier onset of regression and more severe language retardation than patients with a mutation in the methyl-CpG binding domain region. However, the parameters of clinical severity were variable among patients with the same type of mutation, depending on the pattern of X-chromosome inactivation. This study suggests that the X chromosome inactivation pattern can modify the phenotype of Rett syndrome, which is primarily determined by the type and site of MECP2 gene mutation. PMID- 15526955 TI - Study of attentional processes in children with idiopathic epilepsy by Conners' Continuous Performance Test. AB - Before starting antiepilepsy therapy (T1) and after 1 year (T2), 19 children (7 female) affected by focal or generalized idiopathic epilepsy were considered with the aim of studying attention performance. All of the children received a neurological examination and clinical interview, electroencephalography (EEG) in wakefulness or sleep, and a standardized computerized measure of attention (Conners' Continuous Performance Test). At T1, 21% of the patients showed a specific attention disorder, and their percentage rose to 42% after 1 year, despite complete control of seizures. The prognosis was strongly correlated with (1) the presence of active seizures for a period of over 6 months, (2) the persistence of specific interictal abnormalities on an electroencephalogram at T2, and (3) the presence of emotional and behavioral disorders at the diagnosis of epilepsy. Our results confirm that attention disorders are frequent in children with idiopathic epilepsy. The Conners' Continuous Performance Test has proven useful both for diagnosis and follow-up over time. PMID- 15526956 TI - Severe myoclonic epilepsy in infancy: toward an optimal treatment. AB - Severe myoclonic epilepsy in infancy, or Dravet syndrome, is one of the catastrophic epilepsy syndromes. In the past, treatment was mainly based on valproate and phenobarbital. Recently, some of the new antiepilepsy drugs, such as topiramate and stiripentol, have been shown to be promising in the treatment of this epilepsy syndrome. The treatment regimen of 12 children with Dravet syndrome and proven mutations in the alpha subunit of the sodium channel SCN1A is reported here. Five patients on the "traditional" treatment regimen are compared with seven children on an "optimal" treatment regimen based on a combination of valproate and topiramate. With respect to the literature and our own experience, we propose guidelines for "optimal" treatment of children with severe myoclonic epilepsy in infancy. This includes prevention of hyperthermia, rigorous treatment of fever, avoiding stressful situations, maintenance treatment based on a combination of only two antiepilepsy drugs (ie, valproate and topiramate), and a strict acute seizure treatment based on benzodiazepines. To prevent long-lasting periods of status epilepticus, this acute seizure treatment must be taught to parents and caregivers. PMID- 15526957 TI - Familial hyperekplexia and refractory status epilepticus: a new autosomal recessive syndrome. AB - Hyperekplexia is a rare disorder characterized by an exaggerated startle response to noise and handling and by neonatal hypertonia. It is predominantly an autosomal dominant disease; however, atypical cases with additional variable manifestations have been reported. We report a hitherto undescribed association of hyperekplexia and refractory status epilepticus in two siblings. Both children were born after an uneventful pregnancy to healthy unrelated Ashkenazi Jews. Both had increased startle and tone from birth and later became hypotonic. A metabolic evaluation, including a muscle biopsy, was normal. At the age of 18 months and 12 months, respectively, they developed status epilepticus refractory to all treatment that culminated in death. An autopsy in the girl did not reveal any brain pathology. The unusual association of hyperekplexia and refractory status epilepticus in both children suggests that this is a new autosomal recessive syndrome, possibly a channelopathy affecting both the brain and the spinal cord. PMID- 15526958 TI - Cerebral sinovenous thrombosis in children: another reason to treat iron deficiency anemia. AB - Iron deficiency anemia is a rare cause of cerebral sinovenous thrombosis in children. We report three cases of cerebral sinovenous thrombosis and iron deficiency anemia treated at Primary Children's Medical Center in Salt Lake City, Utah, between 1998 and 2001. The children were 9, 19, and 27 months old at the time of admission. Hemoglobin levels ranged from 6.6 to 7.0 g/dL, mean corpuscular volume levels from 45 to 56 fL, and platelet counts from 248,000 to 586,000/microL. Magnetic resonance imaging and magnetic resonance venography revealed thrombosis of the straight sinus and internal cerebral veins in all three children, with the addition of the vein of Galen, left transverse and sigmoid sinuses, and upper left internal jugular vein in one child. Recovery ranged from excellent to poor in 3 months to 3 years of follow-up. Four additional cases, ages 6 to 22 months, were found in the English-language literature. Evaluation for prothrombotic disorders was negative in all children, including the current cases. Treatments have included thrombectomy, corticosteroids, mannitol, heparin, low-molecular-weight heparin, warfarin, aspirin, blood transfusion, and iron supplementation, but there is no consensus regarding therapy, other than to correct the anemia and treat iron deficiency. Iron deficiency anemia, a preventable cause of cerebral sinovenous thrombosis, deserves consideration when cerebral sinovenous thrombosis is detected in young children. PMID- 15526959 TI - Cerebral regional oxygen fluctuations and decline during clinically silent focal electroencephalographic seizures in a neonate. AB - We describe a neonate with tuberous sclerosis complex and right frontal cortical dysplasia who underwent simultaneous near-infrared spectroscopy and electroencephalography (EEG) during repetitive clinically silent right frontal EEG seizures. The seizures produced a progressive decline in regional oxygen saturation index and wider regional oxygen saturation index fluctuations in the right hemisphere than in the left hemisphere. We conclude that recurrent clinically silent focal EEG seizures in this neonate were associated with lateralizing near-infrared spectroscopy changes suggestive of relative cerebral hypoxia. PMID- 15526960 TI - Electroencephalographic correlate of juvenile Huntington's disease. AB - The spectrum of clinical disease in juvenile Huntington's disease differs from that seen in adults. Younger patients often present with seizures, dystonia and rigidity. The mechanism and type of seizures, timing of onset and electrographic features have not been well characterized in either adults or children. We describe the electroencephalographic findings observed in a young child with Huntington's disease who presented with motor regression and seizures. Recordings demonstrated bilateral posterior quadrant epileptiform discharges and occipital intermittent rhythmic delta activity, generally considered a nonspecific abnormality. These findings have not been reported in adult or juvenile Huntington's disease. Its presence in proximity to bilateral posterior spikes suggests that occipital intermittent rhythmic delta activity is an epileptiform abnormality, on a continuum with the posterior spike-and-wave discharges. Intermittent rhythmic delta activity can occur secondary to either the subcortical gray-matter disease or the associated seizure disorder. Huntington's disease should be considered in the differential diagnosis of young children who present with seizures and developmental regression. PMID- 15526961 TI - Lumbosacral agenesis and anteroposterior split cord malformation in a patient with single central maxillary incisor: case report and review of the literature. AB - We report an infant born with lumbosacral agenesis, a subcutaneous lumbosacral mass, and a solitary median maxillary incisor. Operatively, a lipomyelomeningocele was noted to extend from the dorsal hemicord of a split cord malformation. Genetically, the child was found to have a defect on chromosome 7q. To our knowledge, this is the first case report of a patient with the rare lumbosacral agenesis and solitary median maxillary incisor. Furthermore, to our understanding, this is only the fourth reported case of split cord malformation in which the hemicords are oriented in the sagittal plane and one of only a few cases reported in the medical literature of simultaneous occurrence of lumbosacral agenesis and occult spinal dysraphism, that is, split cord malformation and lipomyelomeningocele. Following a review of the medical literature, and considering that our patient did indeed have dysmorphology of chromosome 7q, we hypothesize that this single defect resulted in both cranial and caudal deformation, as exemplified by our patient. We review the medical literature for related case reports and potentially related chromosomal abnormalities and various teratogenic substances. PMID- 15526962 TI - Case of subacute sclerosing panencephalitis with atypical absences and myoclonic atonic seizures as a first symptom. AB - The 2-year history of a 10-year-old boy with subacute sclerosing panencephalitis is presented. After 6 months of epilepsy manifested by atypical absences and myoclonic-atonic seizures with an electroencephalographic (EEG) correlation of generalized spike-and polyspike-wave discharges, the child developed chorioretinitis and demonstrated a dramatic intellectual decline during corticosteroid treatment. Neuroimaging investigations did not demonstrate any pathologic changes, including at the time of fully developed disease, with neurologic deficits, periodic spasms, polymorphic epileptic seizures, and dementia. The typical generalized periodic complexes and general slowing replaced the epileptic abnormalities in the EEG. The initial anticonvulsant treatment was temporarily effective, and the purposeful isoprinosine therapy had no significant beneficial effect. The subacute sclerosing panencephalitis rapidly developed to stages III to IV, and only during the vegetative state did computed tomography show marked brain atrophy. This case is among the few described in the literature with atypical absences and myoclonic-atonic seizures as the first manifestation of subacute sclerosing panencephalitis. PMID- 15526963 TI - Unusual manifestation of subacute sclerosing panencephalitis: case with intracranial high-pressure symptoms. AB - Subacute sclerosing panencephalitis can show variations in clinical course, and some ophthalmologic abnormalities can be seen as cortical blindness and optic atrophy. A 4-year-old girl was referred to our hospital with a complaint of diplopia, vomiting, and ataxia. On physical examination, she was found to have stage IV papilledema with retinal hemorrhage. She was diagnosed as having idiopathic intracranial high pressure until magnetic resonance imaging demonstrated T2-weighted hyperintense lesions. After observation of head drop attacks and detection of elevated antimeasles antibodies in cerebrospinal fluid, the diagnosis of subacute sclerosing panencephalitis was established, and isoprinosine and carbamazepine were started for treatment. However, because carbamazepine failed to control the head drop attacks, topiramate was also included, and the attacks were kept under control with topiramate. The case presented in this article is a good example of subacute sclerosing panencephalitis in which, at early stages, some of the signs and symptoms can lead to an erroneous diagnosis. In addition, we have demonstrated that topiramate might be a good choice for treatment for the persistent myoclonus seen in this type of patient. PMID- 15526964 TI - Lack of long-term histopathologic changes in brain and skeletal muscle of mice treated with a ketogenic diet. AB - Although there is increasing awareness of adverse effects associated with use of the high-fat ketogenic diet, very little is known regarding its long-term clinical consequences, especially in relation to cardiovascular health. Recent reports have highlighted rare but significant cardiac problems in patients treated with the ketogenic diet. Given the inherent limitations in conducting detailed pathologic assessments in patients, we asked whether histologic changes might develop in the brain and skeletal muscle of mice fed a high-fat diet for 2 to 3 months. We found no evidence of gross morphologic or histochemical alterations in muscle or brain after administration of the ketogenic diet. Further, there was no abnormal lipid storage or mitochondrial enzymatic staining. Our data suggest that patients chronically treated with a ketogenic diet are not likely to develop a lipid myopathy or neuronal inclusions. PMID- 15526965 TI - The challenge of neuropsychiatric issues in pediatric epilepsy. AB - Diagnosis and management of pediatric epilepsy are challenging for a variety of reasons; however, one of the most common yet difficult situations involves treatment of the patient with pediatric epilepsy with coexistent cognitive impairment or psychiatric disturbance. The etiologies and presentations of these comorbidities are myriad, making diagnosis problematic; furthermore, interactions between epilepsy and its treatments and the neuropsychiatric problem and its treatments add another layer of complexity to the management of these patients. Antiepileptic drugs are often blamed for cognitive or behavioral problems in children treated for epilepsy, but the actual contribution to such problems in a particular child can be difficult to ascertain. PMID- 15526966 TI - Mechanisms of action for the commonly used antiepileptic drugs: relevance to antiepileptic drug-associated neurobehavioral adverse effects. AB - Antiepileptic drugs exert their anticonvulsant effects by interfering with brain processes that involve structures that are also involved in learning, memory, and emotional behavior. Thus, modulation of ion channels, neurotransmitters, second messengers, and other processes by antiepileptic drugs, although helpful in controlling seizures, can interfere with normal brain function in undesired ways. The specific mechanism(s) of action of an antiepileptic drug can increase the risk for particular types of adverse events. In this review, we examine the cognitive and behavioral effects of antiepileptic drugs in animal models. Although animal studies, in many respects, do not mimic clinical experience, the data suggest a connection between certain mechanisms of antiepileptic action and the occurrence of cognitive adverse effects. Specifically, antiepileptic drugs with traditional gamma-aminobutyric acid (GABA)ergic mechanisms have the most detrimental effects on cognitive function, possibly because they impair attention. Conversely, drugs with the predominant effects at Na+ channels appear to have minimal impact on cognition. Levetiracetam, with its nonconventional GABAergic and Ca2+ channel effects, has shown positive cognitive effects in animal studies. Antiglutamatergic drugs have the potential to be a double-edged sword: they can interfere with consolidation of learning and memory but can also provide neuroprotection in addition to their antiseizure effects. PMID- 15526967 TI - Determining the effects of antiepileptic drugs on cognitive function in pediatric patients with epilepsy. AB - The majority of children with epilepsy are of normal intelligence; however, a significant subset suffers from temporary or permanent cognitive impairment. Factors that affect cognitive function are myriad and include the neuropathology underlying the epilepsy, seizures, epileptiform activity, psychosocial problems, and antiepileptic drug side effects. Although cognitive impairment is often wrongly attributed to the effects of antiepileptic drugs, antiepileptic drugs do impair cognition in some children. Clinicians should be aware of the differential cognitive effects of antiepileptic drugs and should monitor cognitive function closely when adding or changing therapy. Based on published data from prospective, chronic dosing studies, phenobarbital and topiramate have the highest potential for causing cognitive dysfunction. PMID- 15526968 TI - Behavioral and psychiatric adverse events associated with antiepileptic drugs commonly used in pediatric patients. AB - Multiple factors-including the efficacy of the antiepileptic drug for the particular seizure type, availability of pediatrics-friendly formulations, the ease of dosing and titration, and tolerability issues, such as possible drug interactions and adverse events-affect the selection of the best antiepileptic drug for a child with epilepsy. Behavioral problems are common in children with epilepsy and can be aggravated or initiated by antiepileptic drug therapy. The types and frequencies of antiepileptic drug-associated behavioral events can influence a clinician's drug selection, drug management, and counseling of parents. Unfortunately, appreciating differences among antiepileptic drugs in behavioral adverse event profiles is problematic because, among other reasons, methodologies for reporting, collecting, and analyzing adverse events are not uniform across trials, and there is marked heterogeneity in study design among trials. This review summarizes incidence rates for behavioral and psychiatric adverse events taken from studies of children with epilepsy. These rates are reported for the 10 most commonly prescribed antiepileptic drugs (valproic acid, carbamazepine, phenobarbital, lamotrigine, phenytoin, levetiracetam, oxcarbazepine, topiramate, zonisamide, and gabapentin), grouped according to their predominant mechanism of action. Despite the numerous methodologic inconsistencies, some similarities in adverse event profiles among antiepileptic drugs that share mechanisms of action are apparent. Moreover, the overwhelming body of data on the behavioral effects of phenobarbital should convince clinicians that, whenever possible, it should not be used as the initial-or even the second-monotherapy in children with epilepsy. PMID- 15526969 TI - Choosing antiepileptic drugs for developmentally normal children with specific epilepsy syndromes and behavioral disorders. AB - Antiepileptic drugs are often used for the treatment of both epilepsy and a wide range of behavioral and psychiatric disorders. The treatment of patients with epilepsy has been the proving ground for antiepileptic drugs, not only with respect to their efficacy in the treatment of seizures but also for clarifying their dose-related and idiosyncratic adverse events. This information has been useful in treating patients with behavioral and psychiatric disorders. Indeed, the number of prescriptions written for many antiepileptic drugs for nonepileptic uses far exceeds those written for the same drugs for epilepsy. Because patients with chronic epilepsy have a higher incidence of axis I psychiatric disorders, physicians can choose an antiepileptic drug to treat both the epilepsy and psychiatric comorbidity in selected patients. Guided by the principles of evidence-based medicine as outlined by the American Academy of Neurology and the American Academy of Pediatrics, this article reviews the application of antiepileptic drugs for epilepsy and behavioral and psychiatric disorders in children. PMID- 15526971 TI - Management of epilepsy in mentally retarded children using the newer antiepileptic drugs, vagus nerve stimulation, and surgery. AB - Clusters of seizures, prolonged seizures, and status epilepticus occur more frequently in children with multiple disabilities, and chronic seizures are more likely to be refractory to treatment. In many patients, the seizures appear to contribute to the mental retardation. Thus, if the lives of these children are to improve, seizure control is essential. However, medical treatment can interfere with cognition and cause behavioral disturbances, making life very difficult for the child and the child's family. With the introduction of 10 new antiepileptic drugs in the last decade, the treatment of epilepsy in multiply handicapped children has significantly advanced. These new antiepileptic drugs may improve seizure control, medication tolerance, or both. Although the ultimate therapeutic goal is to keep children seizure free and alert, compromises regarding medication choice and dosage are still necessary in many cases. Novel treatment options, such as the vagus nerve stimulator, may decrease seizure frequency without behavioral or cognitive side effects. In carefully selected children with specific epilepsy syndromes, epilepsy surgery can provide partial or complete relief from seizures. PMID- 15526970 TI - Seizures and epilepsy among children with language regression and autistic spectrum disorders. AB - Clinical and subclinical seizures occur frequently among children with autistic spectrum disorders. Electrographic status epilepticus in sleep, or continuous spike-wave in slow-wave sleep, is a typical feature of acquired epileptic aphasia and Landau-Kleffner syndrome. Seizures and epilepsy are more common among children with autistic spectrum disorder who experience language regression, especially those who experience language regression after the age of 2 years. Although the seizures associated with Landau-Kleffner syndrome and with acquired epileptic aphasia can be easily treated, improvement in language function often does not follow successful treatment of seizures. There are no published randomized clinical trials of treatments for Landau-Kleffner syndrome or for autistic language regression. Broad-spectrum antiepileptic drugs not associated with cognitive slowing are probably the treatment of choice for epilepsy among children with autistic spectrum disorder. Large multisite trials are needed to determine treatment efficacy among children with Landau-Kleffner syndrome and acquired epileptic aphasia and to establish whether there is a cause-effect relationship between electrographic status epilepticus in sleep or continuous spike-wave in slow-wave sleep and autistic language regression. PMID- 15526972 TI - Managing behavioral and cognitive problems in children with epilepsy. PMID- 15526973 TI - Expression of integrin fraction and adhesion molecules on human granulosa cells and its relation with oocyte maturity and follicular steroidogenesis. AB - PURPOSE: To study the correlation between the expression of integrin fractions and adhesion molecules on granulosa cells (GC) and follicular development. METHODS: GC and follicular fluid (FF) were obtained at oocyte retrieval for ICSI. Expression of adhesion molecules on GC was studied by flow cytometry. Statistics were evaluated using the Student t test and simple linear regression analysis. RESULTS: alpha5 integrin fraction was significantly (p < 0.01) higher, while alpha6 fraction and CD9 were significantly (p < 0.01 and p < 0.001, respectively) lower in GC from FF with metaphase II oocytes. A direct significant correlation was observed between FF progesterone and the alpha5 expression on GC (r = 0.54). In contrast, an inverse significant correlation was observed between FF progesterone level and the expression of alpha6 and CD9 (r = -0.40 and -0.41, respectively). CONCLUSIONS: The expression pattern for integrin fractions and adhesion molecules may be of predictive value in assessing the state of differentiation of the human follicle. PMID- 15526974 TI - Comparison of clinical outcome and costs with CC + gonadotropins and gnrha + gonadotropins during Ivf/ICSI cycles. AB - OBJECTIVE: To compare clinical outcome and costs of CC + gonadotropins with GnRHa + gonadotropins during IVF/ICSI cycles. MATERIALS AND METHODS: Clinical outcome and expenses of 382 CC + gonadotropin and 964 GnRHa + gonadotropin cycles were compared. Medication costs were calculated on the basis of the mean number of ampoules and the proportion of various gonadotropins. Costs per clinical pregnancy were calculated on the basis of expenses and clinical pregnancy rates. RESULTS: Women in the CC + gonadotropin group were younger, and had fewer follicles, oocytes, embryos, and embryos transferred. Clinical pregnancy rates were higher in the GnRHa group (35.9 % vs 26.2%, p < 0.001). More ampoules of gonadotropins were used in the GnRHa group (24.0 +/- 0.3 vs 20.0 +/- 0.5, p < 0.001). Medication costs per cycle were higher in the GnRHa group (US dollars 357 vs 248). Expenses per pregnancy however were lower in the GnRHa group (USdollars 4197 vs 5335 with IVF; USdollars 5590 vs 7244 with ICSI). When different age subgroups with similar baseline characteristics and stimulation parameters were compared, pregnancy rates were significantly higher in the GnRHa groups. Medication cost per cycle was higher in the GnRHa subgroups, and the expense per pregnancy was lower with GnRHa protocol. CONCLUSIONS: Cost per cycle is higher with GnRHa + gonadotropin. However, because of the better performance of the GnRHa + gonadotropin stimulation, the cumulative costs are reduced by the time a clinical pregnancy is achieved. PMID- 15526975 TI - Improved outcome of frozen-thawed blastocyst transfer with Menezo's two-step thawing compared to the stepwise thawing protocol. AB - PURPOSE: To compare the effects of two different blastocyst thawing protocols on implantation, pregnancy, and live birth rates. METHODS: Ninety four consecutive frozen-thawed blastocyst transfers from 1996 to 2002 were retrospectively analyzed. Blastocysts were cryopreserved using Menezo's two-step slow freezing protocol. Frozen blastocysts were thawed for transfer according to Menezo's stepwise or two-step protocol. Immediately after thawing, assisted hatching was performed and all embryos were cultured for 3 to 5 h before transfer. Only viable embryos were selected for transfer. RESULTS: Implantation, pregnancy (determined by the presence of fetal cardiac activity), and live birth rates were significantly higher with two-step (25, 45.7, and 42.9%) than with stepwise thawing protocol (9.2,18.6, and 16.9%, P < 0.01). The percentage of gestational sacs resulting in live babies was higher with two-step thawing (76.6%) than with stepwise thawing (50%, P = 0.04). CONCLUSIONS: We conclude that the two-step thawing protocol offers advantages over the stepwise method. The two-step thawing protocol dramatically increased embryo implantation potential, resulting in higher pregnancy rate, and subsequent live birth rate, after frozen blastocyst transfer. PMID- 15526976 TI - Impact of body mass index on in vitro fertilization outcomes. AB - PURPOSE: To determine if body mass index (BMI) impacts IVF outcome. METHODS: Retrospective, cohort study. Main outcome measure was number of oocytes obtained. RESULTS: BMI did not correlate with the prestimulation parameters. There was a significant positive correlation between BMI and the number of follicles on ultrasound prior to egg retrieval. A threshold analysis revealed a significant change in parameters at a BMI > 24 kg/m2. Patients with BMI > 24 kg/m2 demonstrated a significant increase in the number of follicles after stimulation (p = 0.03) and a comparative decrease in the number ampules of gonadotropins used (p = 0.04) and days of stimulation required (p = 0.01). CONCLUSION: These data demonstrated that an elevated BMI significantly correlates with the number of follicles, days of stimulation, and number of ampules of gonadotropins used. Further correlation to an actual increase in number of oocytes and pregnancy rates may be limited by insufficient power in this study. PMID- 15526977 TI - Reduced senescence and retained nuclear DNA integrity in human spermatozoa prepared by density gradient centrifugation. AB - PURPOSE: To investigate whether removal of extraneous cells and immotile spermatozoa from a sperm preparation by density gradient centrifugation could help to maintain normal spermatozoa in a viable state and retain their deoxyribonucleic acid integrity. METHODS: Sperm motility was assessed on a daily basis in aliquots of neat semen, extended semen, and spermatozoa prepared on a PureSperm density gradient. At the same time, aliquots of each sperm sample were preserved for TUNEL assay and nick translation. RESULTS: Spermatozoa prepared using density gradient centrifugation survived three times as long as spermatozoa in neat semen or in extended semen. Both deoxyribonucleic acid integrity and sperm motility were retained in the gradient preparations. CONCLUSIONS: Preparing spermatozoa by density gradient centrifugation is advantageous in prolonging sperm survival and maintaining deoxyribonucleic acid integrity, presumably by removing sources of reactive oxygen species. Stored spermatozoa could be used for a second attempt at fertilization if oocyte immaturity was suspected. PMID- 15526978 TI - Coculture with a human granulosa cell line enhanced the development of murine preimplantation embryos via SCF/c-kit system. AB - PURPOSE: To evaluate the effect on the coculture of murine embryos with a human ovarian granulosa tumor derived cell line (KGN cells). METHODS: We observed microscopically the growth of murine preimplantation embryos in the coculture system with KGN cells or in the presence with exogenous stem cell factor (SCF). The reverse transcriptase-polymerase chain reaction (RT-PCR) method was used to analyze the gene expression of SCF in KGN cells cocultured with murine embryos. RESULTS: The coculture system with KGN cells significantly increased the rate of embryo development to late blastocyst and to hatching stage. We also found that coculture with murine embryos enhanced the gene expression of SCF in KGN cells. Adding human recombinant SCF to the medium significantly enhanced embryo development to late blastocyst and hatching stage. CONCLUSIONS: KGN cells may facilitate preimplantion embryo development through SCF/c-kit paracrine system. PMID- 15526979 TI - Spontaneous pregnancies in a Turner syndrome woman with Y-chromosome mosaicism. AB - PURPOSE: To present a case involving pregnancies in a Turner woman with Y chromosome mosaicism. METHOD: A descriptive case report of a single patient. RESULTS: A 39-year-old woman was admitted to the endocrine clinic due to fatigue and premature menopause. She had tried in-vitro fertilization and oocyte donation twice without pregnancies but became spontaneously pregnant at age 36 and 37 and delivered two girls. During the seventh month of the second pregnancy, a dissecting aortic aneurysm, a coarctation, and subsequently a pheochromocytoma were detected and repaired. Hypothyroidism developed. Turner syndrome was diagnosed. Fluorescence in situ hybridization (FISH) analysis of lymphocytes revealed 31% XY cells and 4% XYY cells, while 66% of buccal cells had an XY constitution. Oophorectomy revealed no malignancy. FISH revealed 54% XY cells in the left gonad and 38% XY cells in the right. CONCLUSION: Turner syndrome should be suspected in women with aortic dissection, in general, but especially in those with additional features such as horseshoe kidney, coarctation, and infertility. PMID- 15526980 TI - Comparison of maturation, fertilization, development, and gene expression of mouse oocytes grown in vitro and in vivo. AB - PURPOSE: To investigate the difference of in vitro and in vivo grown oocytes, we compared maturation, fertilization, development, and maternal gene expression from both in vitro and in vivo grown mouse oocytes. METHODS: The preantral follicles isolated from 12-day-old mice were cultured on Transwell-COL membrane inserts. After culture, maturation, fertilization, and developmental rates were assessed. RT-PCR (reverse transcription-polymerase chain reaction) was performed to examine the expression of beta-actin, GDF-9, and IGF-II in matured oocytes. RESULTS: No difference in the nuclear maturation was detected between in vitro and in vivo grown oocytes, but the mean oocyte diameter of the in vitro group was smaller than that of the in vivo group. The fertilization rate was significantly lower in the in vitro group than in the in vivo group (p < 0.05). The capacities of in vitro grown oocyte to cleave and develop to blastocysts were significantly lower than those of the in vivo grown oocytes (p < 0.001). Moreover, blastocyst of in vitro group had fewer total cells than those of in vivo group (p < 0.05). In regards to the expression of genes in mature oocytes, growth differentiation factor-9 (GDF-9) expression was similar between the two groups, but beta-actin was significantly reduced in the in vitro group compared to the in vivo group. Particularly, the expression of insulin-like growth factor II (IGF-II) was not found in the in vitro grown oocytes. CONCLUSIONS: These results showed that in vitro grown oocytes did not have the same developmental capacity as in vivo grown oocytes. We assume that the aberrant expression of maternal-derived genes in the in vitro grown oocytes may cause the poor embryo viability. PMID- 15526981 TI - The presence of cytokines and growth factors in hydrosalpingeal fluid. AB - PURPOSE: To examine the presence of cytokines and growth factors in hydrosalpingeal fluid. METHODS: Eighteen hydrosalpingeal fluids were compared with 15 follicular fluids and serum samples regarding the presence of interleukin 8 (IL-8), IL-12, IL-1alpha, epidermal growth factor (EGF), granulocyte macrophage colony stimulating factor (GM-CSF), leukemia inhibitory factor (LIF), tumor necrosis factor-alpha (TNFalpha), interferon-gamma (IFNgamma), and transforming growth factor-beta2 (TGFbeta2). RESULTS: IL-8 and EGF were detected in all the hydrosalpinx samples. IL-8, IL-12, IL-1alpha, TNFalpha, TGFbeta2, GM-CSF, and LIF were detected to a significantly larger extent in hydrosalpingeal than follicular fluids (p < 0.01). The same cytokines, with the exception of IL-8, TGFbeta2, and LIF, were also more frequently present in comparison with serum. CONCLUSION: The abundant presence of cytokines in hydrosalpingeal fluid suggests an increased expression from the tubal epithelium. Whether high concentrations have a negative influence on embryo development and implantation needs further investigation. PMID- 15526982 TI - Association of inhibin B serum levels with parameters of follicular response in a randomized controlled trial comparing gnRH agonist versus antagonist protocols for ovarian hyperstimulation. AB - PURPOSE: To study the association of inhibin B with ovarian response to FSH stimulation, applying either GnRH agonist or antagonist. METHODS: In a prospective randomized controlled trial, 46 patients undergoing COH received either triptorelin (group I, n = 15) or ganirelix (group II, n = 31). Parameters of follicular response and inhibin B serum levels were assessed. RESULTS: Inhibin B before FSH stimulation was significantly lower in group I than group II. The FSH stimulation phase was significantly longer in group I than group II, and the total FSH dose was significantly higher with a comparable number of retrieved oocytes. Day 1 inhibin B in group I, but not group II, was significantly correlated with the number of large ovarian follicles and retrieved oocytes. In group II, but not group I, inhibin B on day 1 was inversely correlated with the daily and total FSH dose as well as FSH stimulation duration. CONCLUSIONS: The association of inhibin B serum levels with parameters of follicular response in COH is different in patients assigned to GnRH agonist vs. antagonist treatment protocols. PMID- 15526983 TI - The practical implications of a raised serum FSH and age on the risk of IVF treatment cancellation due to a poor ovarian response. AB - PURPOSE: Chronological age, or biological age as indicated by elevated FSH levels, are related to ovarian reserve. This study addresses the likelihood of cancellation of IVF treatment due to a poor ovarian response utilising both basal serum FSH and woman's age. METHODS: A prospective cohort of 536 infertile but ovulating women were studied in their first cycle of IVF treatment. Standardised methods of pituitary desensitisation and ovarian stimulation prior to IVF treatment were employed. Treatment cycles cancelled due to a poor ovarian response to gonadotrophins were studied. A series of logistic regression models were used to explore the probabilities of cancellation in relation to age and FSH. RESULTS: Both age and basal serum FSH levels were independently associated with the risk of treatment cancellation. A low risk of treatment cancellation was observed in women under the age of 35 irrespective of serum FSH, however in older women the risk of treatment cancellation was most likely in women with a high FSH. CONCLUSIONS: In combination both age and FSH may serve as a valuable indicator of poor ovarian response leading to treatment cancellation. However, among older women FSH has particular importance, while less so in younger women with regular menstrual cycles. PMID- 15526984 TI - Does seasonality alter intrauterine insemination outcomes: a 5-year study. AB - PURPOSE: To determine if seasonal changes alter pregnancy rate in intrauterine insemination (IUI) patients. METHODS: One-thousand and eighty semen analyses prepared for IUI were evaluated in this retrospective cohort study of 496 patients. RESULTS: Volume, pH, sperm concentration, and pregnancy rates were not altered by season. However, the percent motility, the total motile spermatozoa in the ejaculate, the straight-line velocity (VSL) of spermatozoa, as well as the morphology of sperm were altered by season. In a subset of these patients that were defined as normal, only the VSL and the morphology of the spermatozoa were altered by seasonal changes. CONCLUSIONS: Seasonality alters sperm motility parameters as well as morphology, but these changes are not significant enough to alter pregnancy rates. PMID- 15526985 TI - Selection of embryos for day-3 transfer at the pronuclear-stage and pronuclear stage cryopreservation results in high delivery rates in fresh and frozen cycles. AB - PURPOSE: Evaluate IVF-ET outcome data for a unique culture and cryopreservation strategy. METHODS: Retrospective study of 92 patients. Embryos for day-3 transfer were selected at pronuclear-stage; all extra zygotes were cryopreserved at pronuclear-stage. RESULTS: Delivery rates for Anonymous Oocyte Donation (Group I), patients <35 years (Group II), and 35-38 years (Group III) were 52.9%, 61.5%, and 51.7% for fresh and 38.5%, 33.3%, and 40.0% for frozen transfer. Deliveries per retrieval were 82.3%, 71.8%, and 58.6%. Only 0.88, 0.80, and 0.61 more zygotes were cultured than what were used for fresh transfer. Singleton, twin, and triplet rates were 64.6%, 31.2%, and 4.2% for fresh and 69.2%, 30.8%, and 0% for frozen. CONCLUSIONS: Selection of day-3 transfer embryos at the pronuclear stage and cryopreservation of extra zygotes results in high delivery rates in fresh and frozen cycles. This approach optimizes deliveries per retrieval and provides many patients with more than one pregnancy per retrieval. PMID- 15526986 TI - FSH time-concentration profiles before and after administration of 0.25 mg cetrorelix in the gnRH-antagonist multiple-dose protocol for ovarian hyperstimulation. AB - PURPOSE: To determine FSH concentration behavior before and after cetrorelix 0.25 mg administration in the GnRH-antagonist multiple-dose protocol on day 6 of ovarian stimulation with 150-300 IU daily recombinant FSH. METHODS: Blood samples for FSH measurements were drawn from seven women every 15 min from 8 h prior to the first cetrorelix administration in the GnRH-antagonist multiple-dose protocol until 15-32 h thereafter. RESULTS: No significant change of FSH concentration was observed. CONCLUSIONS: This observation indicates that no rationale exists of increasing the daily FSH dosage concomitantly to the GnRH-antagonist administration to compensate for a drop of endogenous FSH. PMID- 15526987 TI - Mood and cognitive performance effects of "energy" drink constituents: caffeine, glucose and carbonation. AB - Three studies using healthy volunteers (n = 271) investigated the effects of caffeine, carbohydrates and carbonation in functional "energy" drinks (EDs) with the aim of determining their benefit in every-day life. The results showed caffeine to be the main ED constituent responsible for the effects found, with possible minor, relatively weak effects of carbohydrates. EDs were found to improve and/or maintain mood and performance during fatiguing and cognitively demanding tasks relative to placebo. In terms of absolute values, EDs maintained levels of arousal compared to a deterioration in arousal where placebo was consumed. These effects were found in caffeine-deprived participants, and so may be largely due to "withdrawal reversal". There were only minor differences in the effects of water vs. "sensory-matched" placebo, supporting previous findings indicating that the type of placebo does not alter the conclusions drawn about the effects of the full ED. Finally, carbonation had various effects on mood, some of which were present immediately following consumption, others were consistent with slower absorption of caffeine (and possibly carbohydrates) from carbonated drinks. PMID- 15526988 TI - Prenatal dietary docosahexaenoic acid supplementation in combination with protein restriction does not affect blood pressure in adult Wistar rats. AB - Recent findings indicate that prenatal protein restriction, which leads to elevated blood pressure in adult rats, results in decreased levels of docosahexaenoic acid (DHA) in neonatal rat brain. In light of the evidence of a relationship between dietary DHA and adult blood pressure, the purpose of this study was to ascertain whether prenatal dietary supplementation with DHA would prevent the development of hypertension associated with maternal protein restriction. Throughout gestation, female Wistar rats were fed isocaloric diets containing either 18% casein + 10% corn oil (CON; control), 9% casein + 10% corn oil (LP; low-protein) or 9% casein + 8.5% corn oil + 1.5% DHASCO (LP + 0.6% DHA). DHA increased levels of DHA in neonatal forebrain but there were no effects of LP. At 10 weeks there were no dietary effects on blood pressure measured on four consecutive days using tail-cuff plethysmography. There were also no significant effects measured at 30 weeks, using femoral artery catheterisation, despite adequate power to detect a 10 mm Hg difference. Trends in corticosterone measurements suggested higher stress reactivity in the LP group. These results do not provide strong support for the prenatal low protein model of hypertension and a relation with dietary DHA. PMID- 15526989 TI - Immune response to dietary proteins, gliadin and cerebellar peptides in children with autism. AB - The mechanisms behind autoimmune reaction to nervous system antigens in autism are not understood. We assessed the reactivity of sera from 50 autism patients and 50 healthy controls to specific peptides from gliadin and the cerebellum. A significant percentage of autism patients showed elevations in antibodies against gliadin and cerebellar peptides simultaneously. For examining cross-reaction between dietary proteins and cerebellar antigens, antibodies were prepared in rabbits, and binding of rabbit anti-gliadin, anti-cerebellar peptides, anti-MBP, anti-milk, anti-egg, anti-soy and anti-corn to either gliadin- or cerebellar antigen-coated wells was measured. In comparison to anti-gliadin peptide binding to gliadin peptide at 100%, the reaction of anti-cerebellar peptide to gliadin peptide was 22%, whereas the binding of anti-myelin basic protein (MBP), anti milk, anti-egg and anti-soy to gliadin was less than 10%. Further examination of rabbit anti-gliadin (EQVPLVQQ) and anti-cerebellar (EDVPLLED) 8 amino acid (AA) peptides with human serum albumin (HSA) and an unrelated peptide showed no binding, but the reaction of these antibodies with both the cerebellar and gliadin peptides was greater than 60%. This cross-reaction was further confirmed by DOT-immunoblot and inhibition studies. We conclude that a subgroup of patients with autism produce antibodies against Purkinje cells and gliadin peptides, which may be responsible for some of the neurological symptoms in autism. PMID- 15526990 TI - Malnutrition and REM-sleep deprivation modulate in rats the impairment of spreading depression by a single sub-convulsing dose of pilocarpine. AB - This study aimed to investigate the effect of a single injection of pilocarpine upon the phenomenon of cortical spreading depression (SD), in adult rats submitted to early malnutrition and/or to REM-sleep deprivation for 72h prior to the SD-recordings. The SD was recorded continuously for 3-4h in 13 well-nourished (W) and 15 early-malnourished (M) adult rats. One to two hours after the beginning of the recording session, a sub-convulsing intraperitoneal (i.p.) injection of pilocarpine (190mg/kg) was applied and its effects on SD were studied during the rest of the recording session. Pilocarpine reduced markedly the ECoG amplitudes in all animals and decreased the SD velocity of propagation in the M-, but not in the W-rats, as compared with the pre-drug values for the same animals. In additional 9W- and 10 M-animals, REM-sleep deprivation was induced during the 72 h preceding the SD-recording session. This condition enhanced the pilocarpine effects on SD in the W-, but not in the M-rats, as compared to the respective non-deprived (ND) groups. The results indicate an important acute cholinergic influence on SD, acting by means of pilocarpine activated muscarinic receptors. This effect seems to be differentially modulated by sleep deprivation and malnutrition. PMID- 15526991 TI - Effects of fluoxetine administration on regional galanin expression in obese Zucker rat hypothalamus. AB - The aim of the present work was to study the potential involvement of hypothalamic galanin system in the anorectic mechanism of fluoxetine in obese Zucker rats. Male obese Zucker (fa/fa) rats were administered fluoxetine (10 mg/kg; i.p.) daily for two weeks. The control group was given 0.9% NaCl solution. Significant decreases in food intake, final body weight and total body fat were observed after fluoxetine treatment. Although fluoxetine-treated rats showed a decrease in urine elimination, this effect was not enough to compensate decreased water intake, leading to dehydration, as showed by decreased body water content. Chronic fluoxetine administration increased the numbers of galanin positively immunostained neural cells in medial and lateral preoptic areas, lateral hypothalamic area and paraventricular nucleus (rostral and magnocellular regions), without changes in dorsomedial, ventromedial, supraoptic, suprachiasmatic and arcuate nuclei. Taken into account that galanin stimulates appetite, these results could represent rather a compensatory response against reduced food intake than a direct anorectic mechanism. Changes in the magnocellular region of the hypothalamic paraventricular nucleus suggest a role for galanin neural circuits at this level in fluoxetine-induced hydro-osmotic impairment. PMID- 15526992 TI - Hypothalamic nuclei nitric oxide synthase expression in rats malnourished during early lactation period. AB - In humans and other animals, it has been shown that protein malnutrition during the prenatal period leads to permanent changes, which in adulthood may cause chronic diseases. Molecules involved in the control of energy metabolism could be targets to alterations caused by nutritional status. Some hypothalamic nuclei as the paraventricular (PVN), ventro-medial and arcuate are related to energy metabolism regulation. Orexigenic and anorexigenic molecules are involved in this regulation. Some studies have showed that these nuclei present nitric oxide synthase (NOS) and that it is increased in obese rats. Recently it had been shown that rats malnourished during the lactation period presented metabolic alterations that persist in adulthood. The aim of this work was to study the expression of NOS in hypothalamic nuclei of rats submitted to malnutrition during the early lactation period. Rats from post-natal day (P10) to P90 were used. Control dams were fed with regular chow pellets and diet dams were fed with protein-free chow pellets during the first 10 days of lactation. NADPH-diaphorase or immunostaining techniques were used to access NOS expression in hypothalamic nuclei. Our results show a delay in NOS expression in the PVN and VMH of malnourished rats. It may affect the development of the hypothalamic circuitry, leading to a metabolic imprinting. PMID- 15526993 TI - Effect of adding dietary methionine to a low soy protein diet on the brain protein synthesis rate in ovariectomized female rats. AB - A deficiency of sex hormones affects brain function in mammals, including the decrease of protein synthesis. Recently, we have shown that the protein synthesis in the brain depended on the quality of dietary protein in ovariectomized female rats. The methionine is the first limiting amino acid for the recommended dietary allowance of amino acids in soy protein. The purpose of this study was to determine whether the addition of dietary methionine affected the rate of brain protein synthesis in ovariectomized female rats fed on the soy protein diet. Experiments were conducted on two groups of ovariectomized female rats (24 week) given the diets containing 5% soy protein or 5% soy protein + 0.2% methionine for 10 d. The fractional rates of protein synthesis in cerebral cortex and cerebellum significantly increased with an addition of dietary methionine. In the brain, the RNA activity [g protein synthesized/((g RNA) x d)] was significantly correlated with the fractional rate of protein synthesis. The RNA concentration (mg of RNA/g of protein) was not related to the fractional rate of protein synthesis in any organ. The results suggest that the addition of limiting amino acid for the low soy protein elevates the rate of protein synthesis in the brain of ovariectomized female rats, and that RNA activity is at least partly related to the fractional rate of brain protein synthesis. PMID- 15526994 TI - Delayed onset of ataxia in a patient with short bowel syndrome: a case of vitamin E deficiency. AB - Chronic gastrointestinal disease can result in nutritional deficiencies that can have a direct effect on the neurologic system. Although acute abnormalities can be corrected, symptoms are rarely reversible. Recognizing the appropriate abnormality is a crucial part of long-term treatment strategies in this population. Because motor and cerebellar symptoms can contribute to poor feeding, aggressive supplementation should begin as soon as symptoms are recognized. We present a patient with delayed onset and progressive hypovitaminosis E and briefly review diagnostic and therapeutic options. PMID- 15526995 TI - Michigan plans compete for incentive-laden cap contracts. PMID- 15526996 TI - Michigan health plan sees pluses in new contract. AB - M-CAID in Ann Arbor, MI, uses disease management and pharmacy management to reduce costs, boost quality, and enhance the viability of its capitated contracts. PMID- 15526997 TI - Survey of state programs shows strong reliance on risk contracting. AB - Data Insight. Provider risk sharing in state Medicaid programs appears to be holding steady and even slightly gaining ground compared with a decline in commercial markets. PMID- 15526998 TI - Alta Bates uses hospitalists to reduce inpatient days, improve capitation performance. AB - Hospitalists managing inpatient care at Alta Bates Medical Center hospitals have been able to reduce commercial length of stay to 3.8 days from 6.2 days and Medicare length of stays to 4.2 days from 7.1 since 1999. Here's how..... PMID- 15526999 TI - [Technical plateau in urological consultation]. PMID- 15527000 TI - Pediatric experts make case for providing a 'medical home'. PMID- 15527001 TI - Face-to-face approach proves successful for HIV/AIDS program in Florida. AB - In the past several years, Florida legislators have pinned their hopes on a cadre of DM vendors to bring health care costs under control in that state's Medicaid program. While most of these highly publicized efforts have failed to meet the lawmakers' expectations, the state's HIV/AIDS program has reportedly saved the state as much as dollar 52 million dollars over five years. That program, developed by the Los Angeles, CA-based AIDS Healthcare Foundation, relies on an in-person approach that developers contend is critical to bringing people into care. PMID- 15527002 TI - Innovative technologies offer new tools for DM in rural and underserved areas. AB - The possibilities presented by teleconferencing equipment and Internet-based applications are well-known, but scientists are now looking at the value of wearable devices that can transmit specific data regarding body functions such as energy intake and expenditure, and even sleep. The idea is to arm diabetics, the chronically obese, and disease managers with the kind of real-time information that can influence lifestyle choices when they really count. PMID- 15527003 TI - Interventions show promise in primary prevention of asthma. AB - It's not clear why some children develop allergies and asthma and some children do not, but scientists know that environmental exposures probably account for the epidemic of the disease that has taken place in the developed world over the last three decades. Consequently, a number of researchers are investigating whether environmental interventions can actually prevent onset of the disease. Taking on such an issue for study is not easy, but early results from one investigation in Canada have thus far yielded promising results. PMID- 15527010 TI - Breakthrough to nursing timeline. PMID- 15527011 TI - Touching lives. PMID- 15527012 TI - A care plan for nursing students... 10 antidotes for stress. PMID- 15527013 TI - Strategies for coping with test anxiety. PMID- 15527014 TI - NSNA Leadership University--giving credit where credit is due! PMID- 15527015 TI - Study tips for nursing students. PMID- 15527016 TI - Production of surface active compounds from methane. AB - Methane utilizing bacterial strain was isolated and found to produce a surface active compound while growing on methane. The maximum production of the surface active compound depended on pressure of methane and air in the reactor. The crude surface active compound was extracted from the cell free broth and its surface active and emulsification properties were studied. The purified surface active compound had a critical micelle concentration of 150 mg/l at pH 7.0. On the basis of surface tension reduction, emulsification property and IR-spectrophotometric results this surface active compound was characterized as a glycolipid. PMID- 15527017 TI - Exploring possibilities of achieving sustainability in solid waste management. AB - The solid waste management scenario in the recent years has shifted towards a more sustainable approach. This paper brings in to focus the waste management methods that can be adopted using Bangalore as case study, in order to achieve economic viability and explores the sustainable options that conserves both natural and man-made resources and averts ecological risks. Bangalore, the Garden City of India with a population of 6 million is facing the daunting task of handling 3613 tonnes of municipal solid waste per day. Added to this are the constraints that are faced by the authorities such as poor political back up, inadequate infrastructure, insufficient funds and lack of public support. Attempts have been made to clear wastes by door to door collection method, introduction of push carts, which separate biodegradable wastes from non biodegradable wastes, deployment of auto tippers to clear slum wastes etc. under the guidance of Bangalore Agenda Task Force (government appointed body) and Bangalore Mahanagara Palike (city municipal corporation). Integrated waste management system is proposed as an option, which include collection, transport and processing of wastes in an environmentally sound way. The methods to overcome constraints in waste management and the future plans and actions that will bring about a significant change in the current waste management practices are also discussed in the paper. PMID- 15527018 TI - Arsenic contamination in parts of Yamuna sub-basin, West Bengal. AB - Arsenic rarely occurs in free state and is generally found in combination with Sulphur, Oxygen and Iron. In the environment, Arsenic occurs as a result of several inputs that contain this element in organic and inorganic forms. Arsenic is present in Coal, Lead, Zinc, Gold and Copper ores in the form of several minerals viz. Arsenopyrite (FeAsS), Orpiment (As2S3), Realgar (AsS), Arsenolite (As2O3) and Lollingite (FeAs2). The commonly existing species in groundwater are in two forms viz. Arsenite [As(III)] which is the reduced state of inorganic Arsenic and Arsenate [As(V)], which is the oxidized state of inorganic Arsenic, The As(III) form is more mobile and toxic for living organisms. Recently, the symptoms of chronic arsenic poisoning have been observed in many parts of West Bengal (India) and Bangladesh. In the present study, an integrated evaluation of hydrogeology and hydrochemistry has been carried out by processing the available data of Yamuna sub-basin (Part of Nadia and North 24 Parganas district) of West Bengal in order to understand the possible reason of arsenic pollution in the ground water. Arsenic has generally been observed in higher concentration in shallow aquifer. Northwestern region of the study area has been observed to be the most effected with premonsoon periods exhibiting higher concentration. Transmissivity values have been observed to vary inversely with arsenic concentrations. Arsenic values have not indicated any well defined association with many other constituents. Arsenic buildup could also not be conclusively related to lowering of ground water table of the study area. PMID- 15527019 TI - Influence of Adyar river in the coastal waters of Chennai (Madras), Bay of Bengal. AB - The impact of the river Adyar which is influenced by industrial wastes and domestic sewage from the city of Chennai (Madras), on the hydrobiology of coastal water in the Bay of Bengal was evaluated. Reduction in pH, dissolved oxygen (DO), nitrite, nitrate, primary productivity and increase of ammonia, phosphate, silicate, heavy metals, biological oxygen demand (BOD), chlorophyll pigments, particulate organic carbon (POC), respiration, suspended solids, ATP, and bacteria in the mixing site during low tide indicated that the Adyar water was heavily polluted and ultimately affect the quality of coastal water near the vicinity of mixing zone. The effects are diluted or nullified as the distance increased from the mixing site due to heavy mixing and littoral current. During high tide, the flow was completely stopped and little or no effect was recorded. The light, water current and wave action are the important factors which minimize the pollution impacts in the receiving coastal water. PMID- 15527020 TI - Mapping of fluoride endemic areas and correlation studies of fluoride with other quality parameters of drinking water of Veppanapalli block of Dharmapuri district in Tamil Nadu. AB - 193 drinking water samples from water sources of 27 panchayats of Veppanapalli block of Dharmapuri district of Tamil Nadu were analysed for chemical quality parameters. Based on the fluoride content of the water sources, fluoride maps differentiating regions with high / low fluoride levels were prepared using Isopleth mapping technique. The interdependence among the important chemical quality parameters were assessed using correlation studies. The experimental results of the application of linear and multiple regression equations on the influence of hardness, alkalinity, total dissolved solids and pH on fluoride are discussed. PMID- 15527021 TI - Pyroclastic fluoride in ground waters in some parts of Tadpatri Taluk, Anantapur district, Andhra Pradesh. AB - Fluorosis is a disease affecting the teeth and bones and caused by excessive ingestion of fluoride through drinking water. Fluoride concentration in ground water samples of the study area varies from 1.2 to 2.1 ppm., which is much above the permissible limit. The probable source of fluoride in ground water is fluoride bearing minerals like apatite, micas and clay minerals etc., present in the country rocks like shales and pyroclastic materials of the study area. The sample survey is carried out to know the incidence of fluorosis in this area which reveals that more than 43% of the inhabitants are affected by dental fluorosis and 0.4% by skeletal fluorosis. PMID- 15527022 TI - Environmental monitoring of asbestos products manufacturing units--a case study. AB - Air borne asbestos dust concentration and occupational health environment of workers in a asbestos products manufacturing unit was monitored, and compared with the standards. Study reveals that overall airborne asbestos concentration in the unit is well within the limit, but the workers, which were exposed to air borne asbestos dust, showed a marked increase in deterioration of lung function as compared to the control population, which was not exposed to this dust. Further more, the population which was exposed to airborne asbestos dust along with other predisposing factors like cigarette smoking, showed a marked deterioration of lung function as compared to the population exposed only to air borne asbestos dust. PMID- 15527023 TI - Leaching behaviour of coal-ash: a case study. AB - Leaching of trace elements from fly ash dumps to subsoil layer due to the rain water results in contamination of ground water. The ground water pollution due to fly ash deposition on land so occurring was assessed by simulating the disposal site conditions using two lysimeter with two different soils. Leachate was collected and analysed daily to help understand the phenomenon of leaching of fly ash constituents in the environment. The trace metals and physico-chemical parameters of fly ash and soil used were measured before and after the experiment. Results of analysis of soil and fly ash samples were then compared with the results of lysimeter-I and lysimeter-II. The study reveals that metals respond differently at dumping site while reacting with soil and water. PMID- 15527024 TI - In-vessel composting of different organic waste. AB - Experimental data on bio degradation of wastes with different C/N ratios have been presented for vessel composting conditions. Detailed analysis on the routes of stabilization of organic wastes was done. Greater efficiency during aeration have been pointed out and discussed. Necessity of detailed evaluation of temperature, pH, total solids, E.C, volatile solids, C.O.D, nutrient levels of carbon, nitrogen, phosphorus and potassium, is suggested to achieve quicker bioconversion of organic wastes into humus. Four separate amendment conditions were applied to stimulate varying degrees of bioconversion rate. Optimum environmental conditions were maintained inside vessel through proper aeration, moisture adjustment and sufficient mixing. PMID- 15527025 TI - Major inorganic ion composition of atmospheric aerosols at the coastal site in Visakhapatnam City--India. AB - Atmospheric aerosols were sampled at a coastal site and analyzed for H+, NH+4, Ca++, Mg++, Na+, SO4- -, HCO(-3), Cl- and No3-. The analysis demonstrated that these ions are the main constituents of the aerosols sampled. The correlation coefficients are calculated for each possible pair of anions and cations. Ionic balance in the ambient aerosols indicate that the main compounds existed are in the order of sodium chloride; bicarbonates of calcium and magnesium; sulphates of sodium, magnesium and calcium; and nitrates of sodium, magnesium and calcium. The existence of cations has exceeded the anions which indicate that ionic composition of the aerosols are not very much determined by background marine nature but may also be due to transport of polluted air masses from the surrounding areas. PMID- 15527026 TI - Operating environmental laboratories--an overview of analysis equipment procurement and management. AB - Management of equipment in an environmental laboratory requires planning involving assessment of the workload on a particular equipment, establishment of criteria and specification for the purchase of equipment, creation of infrastructure for installation and testing of the equipment, optimization of analysis conditions, development of preventive maintenance procedures and establishment of in-house repair facilities. The paper reports the results of such an analysis carried for operating environmental laboratories associated with R& D work, serving as an Govt. laboratory or attached to an Industry for analysing industrial emissions. PMID- 15527027 TI - Variation of concentration of heavy metals, calcium and magnesium with sex as determined by atomic absorption spectrophotometry. AB - The concentrations of ten elements: Pb, Cd, Cr, Mn, Fe, Ni, Cu, Zn, Ca and Mg in hair, fingernails and toenails of girl and boy students between 20-24 years of age, living in nonindustrialized areas of Rajasthan state in India were analysed by Atomic Absorption Spectrophotometer. Adequate quality assurance was ensured by inter-laboratory exerciese, the results of which overall showed good consistency. With the exception of Cu, all metals were found to be significant in fingernails and toenails of boys as compared to boys hair while in case of girl students all metals except zinc in toenails were found to be significant in hair. Using Pearson's Correlation Coefficient we obtained information about high positive and negative correlations among metals in hair, finger nails and toenails of boy and girl students. PMID- 15527028 TI - Environmental pollution in rural areas of Orissa state due to industrial emissions--with special reference to fluoride. AB - Angul - Talcher belt in Central Orissa, having a number of industries contributing to a great extent to deteriorate the air quality of the surrounding villages. Previous reports showed higher SPM, SO2, NO, levels in air and prevalence of respiratory illness, skin and teeth disorders among village population. Higher ground water fluoride, urine and serum fluoride among the cattle were also reported in some villages. Present study reports SPM, SO2, NOx and Fluorides (gaseous and Particulate) in ambient air around aluminium smelter during February and August 1996. High volume sampling technique for SPM and the standard colorimetric methods (BIS) for analyses of SO2 and NOx were adopted. Fluoride in air and water were estimated by standard fluoride ion selective electrode method. Higher SPM, SO2 and NOx values than prescribed CPCB standard were obtained in February. Gaseous fluoride in village air were varied between, 1.66 - 7.64 mg/m3 in February and 1.11 - 22.75 mg/m3 in August, whereas particulate fluoride ranged between, 0.054 - 19.61 mg/m3. Water sources of the villages near the smelter showed fluoride values above permissible limit. The study indicated higher fluoride pollution in air and water of the surrounding villages. PMID- 15527029 TI - Efficiency of federal hospitals in the United States. AB - This study evaluates the technical efficiency of federal hospitals in the United States using a variable returns to scale, input-oriented, data envelopment analysis (DEA) methodology. Hospital executives, health care policy-makers, taxpayers, and other stakeholders, benefit from studies that improve the efficiency of federal hospitals. Data for 280 federal hospitals in 1998 and 245 in 2001 were analyzed using DEA to measure hospital efficiency. Results indicate overall efficiency in federal hospitals improved from 68% in 1998 to 79% in 2001. However, based upon 2001 spending of $42.5 billion for federal hospitals potential savings of $2.0 billion annually are possible through more efficient management of resources. From a policy perspective, this study highlights the importance of establishing more specific policies to address inefficiency in the federal health care industry. PMID- 15527030 TI - Classification of mitral insufficiency and stenosis using MLP neural network and neuro-fuzzy system. AB - Cardiac Doppler signals recorded from mitral valve of 60 patients were transferred to a personal computer by using a 16-bit sound card. The power spectral density (PSD) was applied to the recorded signal from each patient. In order to do a good interpretation and rapid diagnosis, PSD values classified using multilayer perceptron (MLP) and neuro-fuzzy system. Our findings demonstrated that 93.33% classification success rate was obtained from MLP, 90% classification success rate was obtained from neuro-fuzzy system. The classification results show that MLP offers best results in the case of diagnosis. PMID- 15527031 TI - Cost accounting by diagnosis in a Japanese university hospital. AB - Cost accounting according to diagnoses covering approximately 600 inpatients with 64 diseases in 20 departments of Kumamoto University was carried out. The reports of these results were automatically generated and used for individual departmental meetings with participating delegates. The administration of each department as well as the management of diseases was discussed at the meetings, and all departments were requested to provide a report of their discussions. We are planning to increase the number of patients in the sample group and to perform more comprehensive and accurate hospital cost accounting. PMID- 15527032 TI - Multimedia based medical instrumentation course in biomedical engineering. AB - Computer assisted instruction in education, including biomedical engineering education, has been explored and changed dramatically for more than two decades. The Internet, with its capacity to transmit synchronous and asynchronous audio, text, and graphics, presents educators with tremendous opportunies for distance education and independent learning. In this work, we have developed a new educational hypermedia for medical instrumentation courses. It is designed to be suitable for biomedical and technical curricula where these courses are scheduled. The courseware provides support for the education of medical instrumentation. The work is presented herein to provide multimedia course material with animations to assist learning some key Medical Instrumentation topics on the World Wide Web. PMID- 15527033 TI - Design and implementation of a calibrated store and forward imaging system for teledermatology. AB - The paper presents a computer-based imaging system aiming to support telemedicine examination sessions in dermatology. Many studies have proved the inadequacy of general practitioners to diagnose successfully common dermatological diseases; some of them may prove fatal if not diagnosed at their early stages (e.g., melanoma). Thus the need for telemedicine systems customized for dermatology becomes obvious for distant rural areas, where dermatological care is usually provided by general doctors. We treat technological issues such as image acquisition, camera calibration, illumination, data transmission, and data compression, and propose a store and forward architecture for image transmission. We also include a study of the effect that image compression quality factor has in the diagnostic value of the skin digital images, along with some initial results and conclusions from the pilot use of the system. PMID- 15527034 TI - Applying a medical budget system to biostatistics clinics. AB - Biostatistics are indispensable in clinical medicine because doctors increasingly need to publish and understand medical articles that use statistical methods. An opportunity is needed for medical doctors to interact with biostatisticians on statistical issues. This paper's objective is to determine how much it costs to establish a biostatistics clinic and what revenue is required to run the clinic. A balance sheet was used to outline the revenue and expenses of a statistics clinic using a simple method introduced by the author. Expenses for personnel, material, and tenancy costs were 41.9, 18.1, and 25.1% of the total, respectively. To run, the biostatistics clinic required 200 research design engagements, 1200 h of hourly consultation, and 6600 h of data processing. To acquire scientific grants, each of four biostatisticians had to write one academic paper annually. The results show that a biostatistics clinic could be managed profitably based on the assumptions in this study. PMID- 15527035 TI - Classification of MCA stenosis in diabetes by MLP and RBF neural network. AB - For the classification of Middle Cerebral Artery (MCA) stenosis, Doppler signals have been received from the diabetes and control group by using 2 MHz Transcranial Doppler. After the Fast Fourier Transform (FFT) analyses of the Doppler signals, Power Spectrum Density (PSD) estimations have been made and Multilayer Perceptron (MLP) and Radial Basis Function (RBF) have been dealt to apply to the neural networks. PSD estimations of Doppler signals received from MCA of 104 subjects have been successfully classified by MLP (correct classification = 94.2%) and RBF (correct classification = 88.4%) neural network. As we have seen in the area under ROC curve (AUC), MLP neural network (AUC = 0.934) has classified more successfully when compared with RBF neural network (AUC = 0.873). PMID- 15527036 TI - An autoregressive (AR) model applied to eye tremor movement, clinical application in schizophrenia. AB - In this study, we use a parametric autoregressive (AR) model to obtain descriptive features of eye tremor movement during fixation. The interest consists in analyzing model parameters to determine the information that can be used as indicator of specific pathophysiology underlying cerebral dysfunction in schizophrenic subjects. We have tested healthy volunteers and schizophrenic medicated and unmedicated patients, to evaluate the treatment effect. The AR model is applied to the eye tremor movement extracted from the eye position signal recorded when subjects are fixating a stationary target. The analysis of the model parameters shows distinct classes, corresponding to a population of subjects among the three kinds included in this study. PMID- 15527037 TI - Report generation and data mining in the domain of thoracic surgery. AB - As a part of AssistMe system, the reporting system has been developed for the thoracic surgery domain. Reporting System is defined as software for dynamic report generation purpose and based on the data-mining techniques. The target users of the future reporting system-physicians, administrative staff, and patients-have been identified. Two major types of clinical reports have been found: predefined and customized. The decision of splitting reports into groups has been taken mainly because users were heterogeneous and had different access rights to the sensitive information. Data-mining process in the reporting system is based on descriptive statistics. It allows dynamically mined AssistMe databases and generates statistical reports about patient's morbidity, mortality, and comorbidity. Information is visualized in the chart way and can be also observed in tabular form. User interaction is also supported by the system. PMID- 15527038 TI - Utilization of public mental health services by adults with serious mental illness. AB - Public mental health (MH) services were examined for non-elderly adults with serious mental illness (SMI) using a database combining information from Medicaid, MH, and substance abuse agencies in three states. These data show that between 23% and 39% of those with SMI received MH services only through Medicaid. Relative use of community versus state hospitals for delivery of psychiatric inpatient care varied across the three states. However, state hospitals accounted for a large proportion of total inpatient days, due to high mean annual days of care. In two states, Medicaid paid for fewer psychiatric inpatient days than expected. PMID- 15527039 TI - Federal demonstration funds at the interface of mental health and criminal justice. AB - A jail diversion program was implemented for offenders with co-occurring mental health and substance abuse disorders (COD) in Lane County, Oregon. This paper discusses whether the program resulted in any sustained community service systems change. Interviewees were asked questions about any systems-level changes that interviewees were aware of in corrections, criminal justice, or treatment where offenders with COD were involved. Results showed evidence of system-level changes in corrections, criminal justice, and treatment that were attributed to various aspects of our "boundary spanning" jail diversion approach with the COD population. The researchers also interpreted the results as evidence of the efficacy of using research to promote change. PMID- 15527040 TI - The responsiveness of State Mental Health Authorities to parents with mental illness. AB - The majority of adults with serious mental illness living in the community are parents, many of whom may be receiving services from State Mental Health Authorities (SMHA). Innovative intervention approaches are available to improve outcomes for these parents and their children. Analyses of SMHA and state-level data, as well as qualitative interviews of administrators, service providers, and consumers, underscore the importance of organizational structure and philosophy, an advocacy presence, and available funding to SMHA efforts on behalf of parents and their families. PMID- 15527041 TI - Mental health, United States, 2002 Executive Summary. PMID- 15527042 TI - Consumer perspectives on services needed to prevent psychiatric hospitalization. PMID- 15527043 TI - Youth substance use and psychosocial well-being in Hungary's post-socialist transition. PMID- 15527044 TI - Comparative flow visualization. AB - There are many situations where one needs to compare two or more data sets. It may be to compare different models, different resolutions, differences in algorithms, different experimental results, etc. There is therefore a need for comparative visualization tools to help analyze the differences. This paper focuses on comparative visualization tools for analyzing flow or vector data sets. The techniques presented allow one to compare individual streamlines and streamribbons as well as a dense field of streamlines. These comparison methods can also be used to study differences in vortex cores that are represented as polylines. PMID- 15527045 TI - Augmented scene modeling and visualization by optical and acoustic sensor integration. AB - In this paper, underwater scene modeling from multisensor data is addressed. Acoustic and optical devices aboard an underwater vehicle are used to sense the environment in order to produce an output that is readily understandable even by an inexperienced operator. The main idea is to integrate multiple-sensor data by geometrically registering such data to a model. The geometrical structure of this model is a priori known but not ad hoc designed for this purpose. As a result, the vehicle pose is derived and model objects can be superimposed upon actual images, thus generating an augmented-reality representation. Results on a real underwater scene are reported, showing the effectiveness of the proposed approach. PMID- 15527046 TI - ISA and IBFVS: image space-based visualization of flow on surfaces. AB - We present a side-by-side analysis of two recent image space approaches for the visualization of vector fields on surfaces. The two methods, Image Space Advection (ISA) and Image-Based Flow Visualization for Curved Surfaces (IBFVS) generate dense representations of time-dependent vector fields with high spatio temporal correlation. While the 3D vector fields are associated with arbitrary surfaces represented by triangular meshes, the generation and advection of texture properties is confined to image space. Fast frame rates are achieved by exploiting frame-to-frame coherency and graphics hardware. In our comparison of ISA and IBFVS, we point out the strengths and weaknesses of each approach and give recommendations as to when and where they are best applied. PMID- 15527047 TI - Image-based collision detection for deformable cloth models. AB - Modeling the natural interaction of cloth and garments with objects in a 3D environment is currently one of the most computationally demanding tasks. These highly deformable materials are subject to a very large number of contact points in the proximity of other moving objects. Furthermore, cloth objects often fold, roll, and drape within themselves, generating a large number of self-collision areas. The interactive requirements of 3D games and physically driven virtual environments make the cloth collisions and self-collision computations more challenging. By exploiting mathematically well-defined smoothness conditions over smaller patches of deformable surfaces and resorting to image-based collision detection tests, we developed an efficient collision detection method that achieves interactive rates while tracking self-interactions in highly deformable surfaces consisting of a large number of elements. The method makes use of a novel technique for dynamically generating a hierarchy of cloth bounding boxes in order to perform object-level culling and image-based intersection tests using conventional graphics hardware support. An efficient backward voxel-based AABB hierarchy method is proposed to handle deformable surfaces which are highly compressed. PMID- 15527048 TI - Interactive volume rendering of thin thread structures within multivalued scientific data sets. AB - We present a threads and halos representation for interactive volume rendering of vector-field structure and describe a number of additional components that combine to create effective visualizations of multivalued 3D scientific data. After filtering linear structures, such as flow lines, into a volume representation, we use a multilayer volume rendering approach to simultaneously display this derived volume along with other data values. We demonstrate the utility of threads and halos in clarifying depth relationships within dense renderings and we present results from two scientific applications: visualization of second-order tensor valued magnetic resonance imaging (MRI) data and simulated 3D fluid flow data. In both application areas, the interactivity of the visualizations proved to be important to the domain scientists. Finally, we describe a PC-based implementation of our framework along with domain specific transfer functions, including an exploratory data culling tool, that enable fast data exploration. PMID- 15527049 TI - Visualization of vector fields using seed LIC and volume rendering. AB - Line Integral Convolution (LIC) is a powerful texture-based technique for visualizing vector fields. Due to the high computational expense of generating the 3D textures and the difficulties of effectively displaying the result, LIC has most commonly been used to depict vector fields in 2D or over a surface in 3D. Here, we propose new methods for more effective volume visualization of three dimensional vector fields using LIC: 1) We present a fast method for computing volume LIC textures that exploits the sparsity of the input texture. 2) We propose the use of a shading technique, called limb darkening, to reveal the depth relations among the field lines. The shading effect is obtained simply by using appropriate transfer functions and, therefore, avoids using expensive shading techniques. 3) We demonstrate how two-field visualization techniques can be used to enhance the visual information describing a vector field. The volume LIC textures are rendered using texture-based rendering techniques, which allows interactive exploration of a vector field. PMID- 15527050 TI - Adaptive extraction of time-varying isosurfaces. AB - We present an algorithm for adaptively extracting and rendering isosurfaces from compressed time-varying volume data sets. Tetrahedral meshes defined by longest edge bisection are used to create a multiresolution representation of the volume in the spatial domain that is adapted over time to approximate the time-varying volume. The reextraction of the isosurface at each time step is accelerated with the vertex programming capabilities of modern graphics hardware. A data layout scheme which follows the access pattern indicated by mesh refinement is used to access the volume in a spatially and temporally coherent manner. This data layout scheme allows our algorithm to be used for out-of-core visualization. PMID- 15527051 TI - Image-space visibility ordering for cell projection volume rendering of unstructured data. AB - Projection methods for volume rendering unstructured data work by projecting, in visibility order, the polyhedral cells of the mesh onto the image plane, and incrementally compositing each cell's color and opacity into the final image. Normally, such methods require an algorithm to determine a visibility order of the cells. The Meshed Polyhedra Visibility Order (MPVO) algorithm can provide such an order for convex meshes by considering the implications of local ordering relations between cells sharing a common face. However, in nonconvex meshes, one must also consider ordering relations along viewing rays which cross empty space between cells. In order to include these relations, the algorithm described in this paper, the scanning exact meshed polyhedra visibility ordering (SXMPVO) algorithm, scan-converts the exterior faces of the mesh and saves the ray-face intersections in an A-Buffer data structure which is then used for retrieving the extra ordering relations. The image which SXMPVO produces is the same as would be produced by ordering the cells exactly, even though SXMPVO does not compute an exact visibility ordering. This is because the image resolution used for computing the visibility ordering relations is the same as that which is used for the actual volume rendering and we choose our A-Buffer rays at the same sample points that are used to establish a polygon's pixel coverage during hardware scan conversion. Thus, the algorithm is image-space correct. The SXMPVO algorithm has several desirable features; among them are speed, simplicity of implementation, and no extra (i.e., with respect to MPVO) preprocessing. PMID- 15527053 TI - Lattice-based flow field modeling. AB - We present an approach for simulating the natural dynamics that emerge from the interaction between a flow field and immersed objects. We model the flow field using the Lattice Boltzmann Model (LBM) with boundary conditions appropriate for moving objects and accelerate the computation on commodity graphics hardware (GPU) to achieve real-time performance. The boundary conditions mediate the exchange of momentum between the flow field and the moving objects resulting in forces exerted by the flow on the objects as well as the back-coupling on the flow. We demonstrate our approach using soap bubbles and a feather. The soap bubbles illustrate Fresnel reflection, reveal the dynamics of the unseen flow field in which they travel, and display spherical harmonics in their undulations. Our simulation allows the user to directly interact with the flow field to influence the dynamics in real time. The free feather flutters and gyrates in response to lift and drag forces created by its motion relative to the flow. Vortices are created as the free feather falls in an otherwise quiescent flow. PMID- 15527052 TI - An intestinal surgery simulator: real-time collision processing and visualization. AB - This research work is aimed toward the development of a VR-based trainer for colon cancer removal. It enables the surgeons to interactively view and manipulate the concerned virtual organs as during a real surgery. First, we present a method for animating the small intestine and the mesentery (the tissue that connects it to the main vessels) in real-time, thus enabling user interaction through virtual surgical tools during the simulation. We present a stochastic approach for fast collision detection in highly deformable, self colliding objects. A simple and efficient response to collisions is also introduced in order to reduce the overall animation complexity. Second, we describe a new method based on generalized cylinders for fast rendering of the intestine. An efficient curvature detection method, along with an adaptive sampling algorithm, is presented. This approach, while providing improved tessellation without the classical self-intersection problem, also allows for high-performance rendering thanks to the new 3D skinning feature available in recent GPUs. The rendering algorithm is also designed to ensure a guaranteed frame rate. Finally, we present the quantitative results of the simulations and describe the qualitative feedback obtained from the surgeons. PMID- 15527054 TI - A clinical study of the involvement of cranial nerves in leprosy. AB - A clinical descriptive study was conducted to assess the frequency and pattern of involvement of cranial nerves in leprosy and to study the relationship of cranial nerve involvement with a leprosy patch or patches on facial skin. One hundred consecutive patients of leprosy, diagnosed by clinical features and/or slit skin smear and histopathology, were studied; of these, 22 patients had cranial nerve involvement. The mean age of patients with cranial nerve involvement was 41.2 years. 16 patients (72.7%) with cranial nerve involvement were in the age-group of 20-49 years. The male-to-female ratio was 3.4:1. The mean duration of the disease in these patients was 5.73 years. The duration of the disease in the majority of patients with cranial nerve involvement was less than 5 years. Impairment of cranial nerves was seen in 12 BT patients, 6 BL patients, and 4 LL patients. No significant difference was noted between involvement of cranial nerves in PB and MB patients. Among the cranial nerves, facial nerve was the most common nerve involved (10/22), followed by olfactory (9/22), trigeminal (7/22) and auditory (3/22) nerves. Among the risk factors, it was found that facial nerve impairment was significantly associated with facial patch(es) and also type 1 lepra reaction. PMID- 15527055 TI - An evaluation of clinical and histopathological status in paucibacillary leprosy patients after completion of fixed duration therapy. AB - The present study was carried out involving 25 patients with paucibacillary leprosy who attended the outpatient department of dermatology of Father Muller's Medical College Hospital during the period January 2001 to March 2002. All the patients were examined clinically and histopathologically at the beginning and at the end of six months of MDT and relevant data recorded. Clinicopathological correlation with histopathological classification before MDT was 72% and 68% at the end of MDT in our study. At the end of treatment 4 (16%) cases were clinically active and 8 (32%) were histopathologlcally active. The study showed that active cases were significantly reduced as a result of MDT, both clinically and histopathologically. The histopathological activity that outlasts MDT may be due to the bacillary fragments that persist; but clinical activity coupled with histopathological activity seen in 2 patients at the end of 6 months of MDT was possibly an indicator of relapse and these patients and similar others need to be followed up for a longer duration. In this study, resolution of granuloma and clinical activity after completion of MDT were assessed. PMID- 15527056 TI - Prevalence of leprosy among household contacts of leprosy cases in western Orissa. AB - The prevalence rate of leprosy among household contacts is an important epidemiological indicator in the character and trend of the disease. During the period January 2001 to December 2002, families of 400 primary cases were studied to detect contact (secondary) cases. Seventy-two cases were detected from 54 families; of these 72 cases, 45 (62.5%) were in the paediatric age-group (0-14 years) and of these 45.8% belonged to 6-14 years group. Out of the 72 cases, there were more (58) of paucibacillary (PNL+I+TT+BT) cases. Secondary cases were significantly high when there was lepromatous type of leprosy in the family. The attack rate among those exposed to paucibacillary type (TT, BT) of leprosy, though much less, was still observed. The influence of duration of contact revealed that a maximum number of cases (N=43, 59.7%) acquired the disease during 0-6 years of contact. A majority of the patients belonged to low-income groups, were illiterate and lived in extended families. The father was the source case for most of the secondary cases (N=41, 57%). Conjugal leprosy was very rare. The results of the study points to the fact that PB leprosy is still a potential source of infection and cannot be ignored. It is important to evolve a hospital based surveillance programme for contacts of leprosy patients since early detection and treatment of contacts are important measures for reducing the reservoir of infection in the community. PMID- 15527057 TI - A study of mast cells in granulomatous lesions of skin, with special emphasis on leprosy. AB - 76 skin biopsies that included material from 7 controls, 65 granulomatous skin lesions and 2 each of granulation tissue and chronic non-specific inflammation, were subjected to histopathological evaluation on haematoxylin and eosin and pertinent special stains. Mast cell study was done on slides stained by toluidine blue method, with special reference to their location, and morphology and cell count were done with the help of occculomicrometre. In normal skin, mast cell density was 11.43/mm2 with a range of 6-22/mm2 and an S.D. of 5.94. Highest value in the whole series was seen in TVC (66/mm2), followed by lupus vulgaris (50/mm2). Mast cell counts were normal in indeterminate and TT leprosy and showed a rise over the immunological spectrum BT to LL, with values in LL being 32.86/mm2 (28-40/mm2). PMID- 15527058 TI - Trends in case detection influenced by leprosy elimination campaigns in certain areas of China. AB - LECs were carried out from 1998 to 2000 in eight counties of west China. The number of cases detected during the year of LECs was much higher than that detected by routine methods before the year of the LEC. However, the annual number of cases detected during the year after the LEC showed different patterns. One pattern is that the number of new cases detected in the year after the LEC declined to the level similar to that before the year of the LEC. The second pattern is that the number of new cases detected in the year after the LEC declined steeply to less than that detected before the year of the LEC. Following peak case-detection during the year of the LEC, a gradual decrease in the number of new cases was observed in the subsequent years. The repeat LEC brought a weakly rebounding peak case-detection during the year following the first LEC carried out 3 years earlier. The operational, epidemiological and technical factors influencing the trends of case-detection during the LECs are discussed. PMID- 15527059 TI - The first international leprosy conference, Berlin, 1897: the politics of segregation. AB - The present paper examines the first attempts to internationalise the problem of leprosy, a subject hitherto overlooked by historians of imperialism and disease. The last decade of the nineteenth century saw many in the civilised countries of the imperialist West gripped by a paranoia about an invasion of leprosy via germ laden immigrants and returning expatriates who had acquired the infection in leprosy endemic colonial possessions. Such alarmists clamoured for the adoption of vigorous leper segregation policies in such colonies. But the contagiousness of leprosy did not go unquestioned by other westerners. The convocation in Berlin of the first international meeting on leprosy revealed the interplay of differing and sometimes incompatible views about the containment of leprosy by segregation. The roles of officials from several countries, as well as the roles of five protagonists (Albert Ashmead, Jules Goldschmidt, Edvard Ehlers. Armauer Hansen, and Phineas Abraham) in the shaping of the Berlin Conference are here examined. PMID- 15527060 TI - Involvement of genitofemoral nerve with genital lesions in lepromatous leprosy. AB - The case of a male patient diagnosed to have lepromatous leprosy with type 2 reaction on multibacillary multidrug therapy, with unusual, widespread involvement of genitalia in the form of plaque and nodules of leprosy over scrotum and perimeatal region of glans, necrotic lesions of erythema nodosum leprosum over scrotum, neuritis of genital branch of genitofemoral nerve bilaterally, and azoospermia, is reported. PMID- 15527061 TI - Cryoglobulins and rheumatoid factor in leprosy. PMID- 15527062 TI - Leprosy situation in endemic states of India and prospects of elimination of the disease. PMID- 15527063 TI - Direct effects of high concentrations of dimethylsulfoniopropionate, vitamin E and ferulic acid on the senility of aged scenescene-accelerated mouse (SAMP8). AB - The effects of high concentrations of dimethylsulfoniopropionate (DMSP), vitamin E and ferulic acid solutions on the aging of male and female scenescene accelerated mouse (SAMP8) at the age of 50 wk were examined by direct supplementation to their stomachs twice a week for 28-30 d. The addition of 1 mL of DMSP, vitamin E and ferulic acid solutions (21% each) to the male and female mice in each group in this order rather elevated their growth and significantly suppressed their total grading score and loss of learning and memory with increasing rearing times for the short experimental period. However, there were no significant differences found between the male and female mice during the experimental period. The antioxidant and hormonal actions for the effects of the test compounds on the aged SAMP8 were possibly considered. PMID- 15527064 TI - Folate intake, serum folate, serum total homocysteine levels and methylenetetrahydrofolate reductase C677T polymorphism in young Japanese women. AB - Various factors influence folate status and requirements. To investigate the folate status in young Japanese women (n=340, 20-22 y), I determined the serum folate and total homocysteine (tHcy), and examined the influence of methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism on folate status in a subgroup of 252 of the women. I calculated folate consumption based on records of food intake weighed over 3 d using the standard Tables of Food Composition in Japan (5th revised edition) and the US Food Composition Table based on the USDA nutrient database excluding fortification. Serum folate and total homocysteine (tHcy) levels were determined by chemiluminescent-competitive protein binding assay and HPLC, respectively. The mean intake of folate was 323+/ 133 and 196+/-71 microg/d, and 86.5% and 42.1% of the participants consumed more than the RDA of 200 microg of folate, according to the Japanese and US tables, respectively. The main sources of dietary folate were vegetables, green tea and bread. Serum folate levels were significantly correlated with folate intake (r=0.249, p<0.001). Serum tHcy levels were above 15 micromol/L in four participants. I found significant negative correlations between serum tHcy and folate intake (r=-0.175, p<0.05), and between serum tHcy and serum folate (r= 0.297, p<0.0001). The frequencies of the CC, CT and TT genotypes of the MTHFR gene were 32.9, 51.6 and 15.5%, respectively. Among individuals with the TT genotype, serum folate and tHcy levels were lower and higher, respectively, than in those with the other genotypes, regardless of folate intake above RDA. These data suggest that the current RDA of folate estimated based on foreign data might not be sufficient for Japanese and that this value might require reassessment. PMID- 15527065 TI - Blood isoflavone levels during intake of a controlled hospital diet. AB - Isoflavones are reported to have an estrogenic activity to prevent prostate cancer. Therefore, it is necessary to analyze the factors influencing the absorption and metabolism of isoflavones using detailed and precise dietary information. We evaluated the relationship between the amount of intake of soybean isoflavones in the diet and its serum levels in 88 hospitalized patients (22 with prostate cancer, 66 without cancer) with a mean age of 67.0+/-9.3 y. The intake amount of genistein and daidzein was significantly lower in the hospital diet than that in the ordinary daily diet. The serum levels of isoflavones were related to the amount of intake consumed during the day before blood collection (r=0.27 for genistein, r= 0.33 for daidzein), but not to the last meal before blood collection. While little relationship was observed between the amount of intake and serum levels of isoflavones in the equol producers, a marked relationship was noted in the non-producers (r=0.52 for genistein, r=0.67 for daidzein). Blood isoflavone levels decreased when the duration of the hospitalization was longer than 1 wk. These observations indicate that the serum isoflavone levels are correlated with the amount of intake of isoflavones during the preceding 2 wk and serve as an effective biomarker in individuals during the intake of the hospital diet. Further investigations, including the mechanism of the metabolism on isoflavones, are necessary for the evaluation of the preventive effect of isoflavones. PMID- 15527066 TI - The effects of bitter melon (Momordica charantia) extracts on serum and liver lipid parameters in hamsters fed cholesterol-free and cholesterol-enriched diets. AB - The hypolipidemic effect of dietary methanol fraction (BMMF) extracted from bitter melon (Koimidori variety), at the levels of 0.5% and 1.0%, was examined in male golden Syrian hamsters fed diets supplemented with and without cholesterol. The feeding of BMMF at 0.5% and 1.0% levels in the diets for 4 wk tended to reduce food intake and growth, although there was no difference in food efficiency (weight gain/food intake). An effect of dietary BMMF on serum triglyceride was not seen in hamsters fed diets free of cholesterol, while hypertriglyceridemia induced by dietary cholesterol was significantly lowered in a dose-dependent manner in those fed diets containing the BMMF Serum total cholesterol concentration also tended to decrease in a dose-dependent manner following feeding of increasing amounts of BMMF in the presence and absence of cholesterol in the diet. The effects of dietary BMMF on liver triglyceride and total cholesterol levels were marginal, although dietary cholesterol caused a marked accumulation of these lipid molecules in the liver. These results suggest that the BMMF contains some components that could ameliorate lipid disorders such as hyperlipidemia. PMID- 15527067 TI - Methionine supplementation did not augment oxidative stress, atherosclerotic changes and hepatotoxicity induced by high cholesterol diet in C57BL/6J mice. AB - The purpose of this study was to investigate the effect of a high-methionine plus cholesterol diet (HM+HC) on plasma, erythrocyte, liver and aorta lipid, lipid peroxide levels, and the liver antioxidant system, as well as hepatic and aortic histopathology in CS 7BL/6J mice, and to compare these results to those observed following administration of a high-methionine (HM) or high-cholesterol diet (HC) alone. Mice were fed diets containing 1.5% methionine, 1.5%, cholesterol and 0.5% cholic acid, or a combination of the two diets, for 4 mo. The HM diet did not alter cholesterol or diene conjugate (DC) levels in the plasma or aorta, but this diet caused increases in cholesterol, triglyceride, malondialdehyde (MDA) and DC levels and a decrease in a-tocopherol levels without any change in the levels of glutathione and ascorbic acid or the activities of superoxide dismutase, glutathione peroxidase and glutathione transferase in the liver of mice. However, the HC diet alone was found to further increase cholesterol, triglyceride. MDA and DC levels in the plasma and liver together with changes in hepatic antioxidant system elements, but aortic cholesterol and DC levels remained unchanged as compared to the control group. There were no changes in blood hemoglobin and erythrocyte MDA levels or erythrocyte hemolysis values in both the HM and HC groups. However, the parameters related to lipid and lipid peroxide and antioxidant systems did not change in the plasma or tissues of the HM+HC and HC groups. Only plasma cholesterol was observed to increase in the HM+HC group as compared to the HC group. In addition, histopathological findings in the liver and aorta were similar in the HC and HM+HC groups. In conclusion, our results indicate that the addition of methionine to the HC diet did not augment oxidative stress, hepatotoxicity or atherosclerotic changes induced by the HC diet in mice. PMID- 15527068 TI - The specific expression patterns of lactase, sucrase and calbindin-D9k in weaning rats are regulated at the transcriptional level. AB - During weaning, rat lactase-phlorizin hydrolase (LPH) expression decreased to the low levels found in adults, while sucrase-isomaltase (SI) sharply increased. Calbindin-D9k (CaBP) is specific to the intestine and expression peaked within a few days of weaning. The present study investigates whether these molecules are regulated at transcriptional or post-transcriptional levels and examines the effects of diet on regulation. At normal weaning on day 21, litters were separated from their dams and one group was fed with a standard laboratory diet (weaned (W) group). The other group received a diet containing lactose as the sole source of carbohydrate (lactose-fed (L) group). Mucosal cells were obtained from the proximal part of the rat small intestine and then the activity and concentration of LPH, SI and CaBP proteins and mRNAs were determined. Three parameters revealed the same changing patterns in LPH, SI and CaBP during development and there was significant (p<0.001) correlation between three parameters: LPH, r=0.97 for activity vs. protein, r=0.99 for activity vs. mRNA, r=0.96 for protein vs. mRNA, SI, r=0.99 for activity vs. protein, r=0.98 for activity vs. mRNA, r=0.96 for protein vs. mRNA, CaBP, r=0.94 for activity vs. protein, r=0.97 for activity vs. mRNA, r=0.95 for protein vs. mRNA. Expression of the three proteins did not differ between the L and W groups. Accordingly, it has been suggested that the expression of LPH, SI and CaBP during development is defined at the transcriptional level and dietary changes do not exert a primary effect on it. PMID- 15527069 TI - Alleviation of fatty liver by alpha-linolenic acid. AB - We compared the efficacy of alpha-linolenic acid (alpha-LNA, n-3) and linoleic acid (LA, n-6) on orotic acid (OA)-induced fatty liver in Sprague-Dawley rats. Rats were fed semi-synthetic diets containing either LA or alpha-LNA with or without 1% OA for 2 wk. OA supplementation lowered serum lipids in LA+OA groups. In addition to the decline of serum lipids in alpha-LNA groups compared to LA groups, a further decrease was found in alpha-LNA+OA groups compared to LA+OA groups. OA-containing diets significantly increased the liver weights and triacylglycerol (TG) accumulations compared with the OA-free diets. These results were attributed to the significant increases in the activities of phosphatidate phosphohydrolase (PAP), a rate-limiting enzyme of TG synthesis, and glucose-6 phosphate dehydrogenase, a fatty acid synthesis-related enzyme. However, the increase of PAP activity was significantly less in the alpha-LNA+OA group as compared with the LA+OA group. These results suggest that dietary alpha-LNA alleviates OA-induced hepatic TG accumulation through the attenuation of hepatic TG synthesis in rats. PMID- 15527070 TI - Gamma-tocopherol enhances sodium excretion as a natriuretic hormone precursor. AB - Endogenous natriuretic factors are believed to be responsible for extracellular fluid homeostasis in mammals. A new endogenous natriuretic factor, Loma Linda University-alpha (LLU-alpha) has recently been proven to be a 2,7,8-trimethyl-2 (2'-carboxyethyl)-6-hydroxychroman (gamma-CEHC), which is a metabolite of gamma tocopherol (gamma-Toc). The purpose of this study was to investigate whether gamma-Toc could accelerate sodium excretion into rat urine as a natriuretic hormone precursor. Male SD strain rats were divided into two groups; one was a control diet group, while the other was a high NaCl group (50 g/kg diet). Next, the two groups were each subdivided into two groups consisting of a placebo group and a gamma-Toc group. After the oral administration of one experimental dose of 20 mg gamma-Toc or placebo, rat urine was collected at 6 h intervals for 24 h, and then the urine volume, sodium and potassium and gamma-CEHC content were determined. gamma-Toc increased in the urine volume of the high-NaCl intake group. The sodium excretion in the high-NaCl group given gamma-Toc was 8.29+/ 2.20 g, while in the control group given gamma-Toc it was 6.24+/-1.49 g from 12 18 h. In contrast, the potassium excretion in the rat urine did not change in any of the groups. Our findings suggested that gamma-Toc accelerates the degree of sodium excretion in rats with a high sodium intake. PMID- 15527071 TI - The effects of carbohydrate consumption on stress levels in humans. AB - A significant increase in calculation ability was observed in the glucose, fructose and sucrose administration groups during both the first and second half of the Uchida Kraepelin test. There was no significant difference in alpha waves between carbohydrate groups (glucose, fructose and sucrose) and control groups at baseline. After a stress situation, there was a significant increase in alpha waves at 11-20 min (p<0.05) in glucose, 11-20 min (p<0.01) and 21-30 min (p<0.01) in fructose, and 0-10 min (p<0.05) in sucrose compared, with each control group. There was no significant difference in beta waves in any of the paired groups. PMID- 15527072 TI - Uptake of iron by a vegetable; kaiware daikon (Japanese radish sprout). AB - Uptake of iron and several other minerals into kaiware daikon (Japanese radish sprout) was investigated by changing their supplementary conditions. The supplementation of calcium sulfate (0.1% (w/v), 230 ppm as calcium ion) and iron (III) chloride (0.1% (w/v), 200 ppm as iron ion) in hydroponics promoted the growth of kaiware daikon ca. 1.0-1.5 cm longer than the control group of no additions, while magnesium sulfate (0.1% (w/ v), 100 ppm as magnesium ion) did not affect the length of sprouts so much, but caused their leaves to grow a little wider. Mineral contents in the sprout were maximal when it was added, at ca. 0.1% (w/v), to the hydroponic solution. In tests regarding iron (II) ion supplementation, the color of the kaiware daikon seeds changed from brownish to black and the sprout did not grow well. In contrast, the iron (III) ion did not affect their color and promoted growth of the sprout. When the seeds were soaked in an iron (II) sulfate (75 ppm) or an ammonium iron (III) citrate (180 ppm) solution for 24 h prior to germination, the resulting kaiware daikon contained a few times higher the amount of iron than commercially available ones. PMID- 15527073 TI - Saccharification of Okara fiber by plant dietary fiber hydrolases. AB - In this paper, the saccharification process of okara fiber with pectinase, xylanase and cellulase was investigated as a preliminary attempt to effectively utilize okara fiber. The solubilization of okara fiber was raised in proportion to the number of enzymes used; that is, by a single enzyme, two enzymes and three enzymes, in that order. The saccharification of okara fiber was much more completed by a combination of pectinase, xylanase and cellulase than by using individual preparations. This multi-enzyme system liberated sugars equivalent to 80% of the original okara fiber by weight. Moreover the structure of okara fiber gradually disintegrated by hydrolytic reactions with pectinase, xylanase and cellulase in turns. These results suggest that the saccharification of okara fiber progresses by the cooperative action of pectinase, xylanase and cellulase. PMID- 15527074 TI - Hot-water extracts from adzuki beans (Vigna angularis) suppress not only the proliferation of KATO III cells in culture but also benzo(a)pyrene-induced tumorigenesis in mouse forestomatch. AB - Treatment of human stomach cancer KATO III cells with hot-water extracts from adzuki beans led to their growth inhibition as well as apoptosis induction. There are morphological changes in the cultured cells treated with the extracts, by which DNA fragmentation characteristic of apoptosis was actualized both concentration- and time-dependently. In contrast, N-acetyl-L-cysteine suppressed such DNA fragmentation, implying that the extracts from adzuki beans might exert antitumorigenicity via active oxygen-induced apoptosis. In order to verify this hypothesis in animal experiment, the 40% ethanol fraction of hot-water extracts was examined for its preventive effect against benzo(a)pyrene-induced tumorigenesis in the forestomach of A/J mice, given as drinking water containing the above fraction at 0.5-2.0% levels. Consequently, forestomach cancer has turned out to be reduced by 36-62% in tumor weight relative to the control. These results suggest that the fraction of hot-water adzuki extracts may serve as a nutrapharmaceutical or functional food available for cancer prevention. PMID- 15527075 TI - Reductive effect of hot-water extracts from woody ear (Auricularia auricula-judae Quel.) on food intake and blood glucose concentration in genetically diabetic KK Ay mice. AB - Genetically diabetic (type II) KK-Ay mice, male and 5 wk of age, were divided into four groups and fed test diets containing 2 (2F), 5 (5F), 10 (10F), or 20 (20F)% fat for 4 wk, and then the 5F group and one-half of the 20F group continued to be fed the corresponding diet and the other half of the latter group was given the 20F (20FHWE) diet with 5% hot-water extracts (HWE) from woody ear (Auricularia auricula-judae Quel.) for a further 3 wk. Remarkable changes in body weight were unfound among the dietary groups. The food intake generally decreased according to the increase in dietary fat content, but the energy expenditure was hardly different among the dietary groups except for the 20FHWE group. That of the 20FHWE group was further decreased compared to the 20F group. Water consumption dropped in the order of the 20F, 5F, and 20FHWE groups. The plasma glucose concentration was the highest in the 20F group, followed in order by the 10F and 5F groups, and the lowest in the 2F and 20FHWE groups. The contents of dietary fat were most positively correlated with the plasma glucose level. The present results using KK-Ay mice confirm that dietary fat levels control postprandial glycaemia, and suggest that the hypoglycemic effect of HWE is primarily caused by a reduction in food ingestion. PMID- 15527077 TI - Chairman's reflection on the past, present and future of neurosurgical oncology. AB - The development of neurosurgical oncology as a subspecialty is closely tied to the development of neurosurgery as a whole. Therefore, the progress that has taken place in the diagnosis and the surgical management of neurosurgical disorders has been widely applied to oncologic disorders affecting the nervous system. The challenges and opportunities that characterize the specialty are grouped into five general categories. These are issues that are related to (1) the management of a large and diverse patient population, (2) the proper training of neurosurgeons to develop the set of required technical skills, (3) the lack of disease curability and its associated opportunity for research endeavors, (4) the importance of the multidisciplinary coordination of care in a horizontal matrix model, and (5) the psychosocial dimensions that are a part of the complexity of human nature. Based on a current assessment of the subspecialty, a perspective on the unfolding future is obtained. This future can be characterized by a stronger workforce, a broader connectivity and representations, and an improved scientific inquiry. PMID- 15527076 TI - History of the AANS/CNS joint section on tumors and preface to the 20th anniversary Journal of Neuro-Oncology Special Issue. AB - The Joint Section on Tumors of the American Association of Neurological Surgeons (AANS) and the Congress of Neurological Surgeons (CNS) was formed in 1984, at the suggestion of Dr. Edward R. Laws, Jr. and with Dr. Mark Rosenblum as the first Section Chair. The Joint Section on Tumors is the first professional organization devoted to the study and treatment of brain tumors. Its initial goals were to 'assist in the education of neurosurgeons in neuro-oncology and to serve as a resource [to the AANS and CNS] and other national groups on the clinical treatment of and research into nervous system tumors'. During its 20-year history, the Section has facilitated both open and invited talks at the neurosurgical national meetings, conducted its own Satellite Symposia, and instituted multiple awards and grants. Members have conducted research surveys and national practice pattern studies, and have collected and disseminated information on clinical protocols, research funding opportunities, and fellowships in neurosurgical oncology. Guidelines for brain tumor treatment and for neuro-oncology fellowships for surgeons have been written by Section committees. Studies presented orally at Section meetings, 1999-2002, had a remarkably high rate of full publication compared to other meetings - 73% actuarial at 4 years after presentation. Finally, nationwide in-hospital mortality rates for craniotomy for malignant glioma have fallen from 8 to 2% during the Section's existence. These data suggest that the Section's goals of educating all surgeons in neurosurgical oncology are being successfully met. A bibliography of secondary sources on the history of brain tumor surgery is appended. PMID- 15527078 TI - The modern brain tumor operating room: from standard essentials to current state of-the-art. AB - It is just over a century since successful brain tumor resection. Since then the diagnosis, imaging, and management of brain tumors have improved, in large part due to technological advances. Similarly, the operating room (OR) for brain tumor surgery has increased in complexity and specificity with multiple forms of equipment now considered necessary as technical adjuncts. It is evident that the theme of minimalism in combination with advanced image-guidance techniques and a cohort of sophisticated technologies (e.g., robotics and nanotechnology) will drive changes in the current OR environment for the foreseeable future. In this report we describe what may be regarded today as standard essentials in an operating room for the surgical management of brain tumors and what we believe to be the current 'state-of-the-art' brain tumor OR. Also, we speculate on the additional capabilities of the brain tumor OR of the near future. PMID- 15527080 TI - Surgical resection for patients with benign primary brain tumors and low grade gliomas. AB - 'Benign' brain tumors are a heterogeneous group of primary neoplasms representing 40% of primary brain tumors. Amongst neuroepithelial tumors, astrocytic neoplasms predominate, followed by oligodendroglial and mixed glial and glial-neuronal neoplasms. Less common are tumors of the ependyma, choroid plexus and neuroepithelial tumors of unknown origin such as chordoid glioma of the third ventricle. Neurosurgical intervention is indicated in order to establish a diagnosis, alleviate symptoms of mass effect, hydrocephalus, or hemorrhage, remove a seizure focus and decrease the cell pool at risk of malignant degeneration. As the majority of patients survive their tumor for years after initial diagnosis, the benefits of therapy must be carefully weighed against adverse treatment effects. This article, with a particular focus on surgical management, reflects the authors approach to the treatment of low-grade tumors of the brain parenchyma in the adult and reviews the pertinent literature published on this controversial issue. PMID- 15527079 TI - The changing role of stereotaxis in surgical neuro-oncology. AB - PURPOSE: Evaluate evolution and time course of stereotactic neurosurgery within surgical neuro-oncology. METHODS: MEDLINE search 1966-2003 sub-stratified and analyzed for annual trends. AANS/CNS membership databases for Joint Sections. ACRC neuro-oncology program database 1998-2003. RESULTS: Tumor stereotaxis emerged in 1980 and became the dominant stereotactic publication topic by 1984. Frame-based tumor stereotaxis led publications through 1994, when supplanted by stereotactic radiosurgery (SR). Brachytherapy led SR 1982-1987, but then fell behind, reducing to pre-1983 levels by 1996. SR publications currently comprise 65% of stereotactic tumor articles and publication rate continues to rise at a steady rate. Frameless stereotaxis (FS) publications began to increase in 1993 and growth is larger than the corresponding fall in frame-based volumetric resection publications. Data suggest increased utilization for cases that would have otherwise utilized ultrasound or gone without image guidance. Intraoperative MR developed predominantly as complimentary technology to FS. Tumor diagnostic needle biopsy publications continue to be mostly frame-based, while FS techniques are largely resection focused. This may change as >80% of our tumors biopsied with frame-based techniques would be candidates for FS biopsy based solely on lesion size, location, and technique accuracy considerations. CNS parenchymal delivery of experimental therapies continues to be predominantly frame-based. CONCLUSION: The role of tumor stereotaxis in surgical neuro-oncology is important, but changing. SR is increasingly dominating the subspecialty. Stereotactic tumor resection has become a mainstream neurosurgical procedure due to FS, and this will likely occur with needle biopsy as well. Delivery of experimental therapies remains predominantly frame-based, but may need to transition to FS in order to gain wider mainstream acceptance and applicability once efficacy is demonstrated. PMID- 15527081 TI - Microsurgery for malignant gliomas. AB - Reflections directly involving the ongoing, intense research activities in biology, the neurosciences and in neurosurgery are discussed including the evolving diagnostic and treatment modalities of primary and secondary malignant gliomas of the central nervous system. The etiology of this enigmatic disease remains obscure, and a curative therapy is still not available. Nevertheless, as a result of changing paradigms in neuroanatomy, neuropathology, neurophysiology, neuroradiology and in neurosurgery, and taking into account the broader selection of adjuvant therapies available, well circumscribed malignant gliomas, which are in predilected compartments of the brain, can be efficiently resected. Good life quality and a respectable survival time are achieved in the majority of patients. PMID- 15527082 TI - Stereotactic radiosurgery and interstitial brachytherapy for glial neoplasms. AB - The application of focal radiation therapies in the management of malignant gliomas has gone through a number of stages. Earlier efforts to improve local control of malignant gliomas involved the use of brachytherapy. Despite some early encouraging results, Phase 3 studies did not prove a significant survival benefit for the addition of brachytherapy for newly diagnosed glioblastoma. Most recently radiosurgery has been employed using the same rationale in that improved local control may improve survival. Results of the RTOG Phase 3 study are pending final publication, but early abstracted reports are negative. While radiosurgery and brachytherapy continue to be used as a form of therapy for selected patients with recurrent gliomas, new information from metabolic imaging studies suggests our problem with these techniques in part may be related to targeting. This paper reviews the recent literature and results of the use of brachytherapy and radiosurgery in the management of newly diagnosed and recurrent malignant gliomas. PMID- 15527084 TI - Surgical resection for patients with solid brain metastases: current status. AB - Brain metastases occur in up to 40% of patients with cancer. Their management has been revolutionized in the last decade by three developments: improved imaging and detection of metastases, better treatment of systemic disease with the result that metastases occur more often; and improved surgical techniques including image-guided surgery to treat metastatic lesions. Class 1 data suggest that surgery is a better treatment for metastases than whole brain radiation. Other data suggest that metastases even in eloquent cortex can be removed safely. The complication rate is low and the recurrence rate is less than 10%. In general, indications for surgery include a mass with an unknown primary; a symptomatic mass including one in eloquent areas; a mass with considerable edema requiring high dose steroids; a mass greater than 3 cm; or patient preference when radiosurgery may also be an option. The question of radiosurgery or whole brain radiation as adjunct to surgical removal requires further evaluation. PMID- 15527083 TI - Neurosurgical delivery of chemotherapeutics, targeted toxins, genetic and viral therapies in neuro-oncology. AB - Local delivery of biologic agents, such as gene and viruses, has been tested preclinically with encouraging success, and in some instances clinical trials have also been performed. In addition, the positive pressure infusion of various therapeutic agents is undergoing human testing and approval has already been granted for routine clinical use of biodegradable implants that diffuse a chemotherapeutic agent into peritumoral regions. Safety in glioma patients has been shown, but anticancer efficacy needs additional refinements in the technologies employed. In this review, we will describe these modalities and provide a perspective on needed improvements that should render them more successful. PMID- 15527085 TI - Stereotactic radiosurgery for patients with solid brain metastases: current status. AB - The goal of this article is to provide a contemporary update on the use of stereotactic radiosurgery (SRS) for the treatment of intracranial metastatic disease. We discuss the rationale for employing SRS in brain metastases and describe the critical factors that predict outcome. We highlight the main clinical indications for SRS including treatment of recurrent brain metastases after previous whole brain radiation therapy (WBRT), as a boost after WBRT, and as sole therapy for newly diagnosed tumors. For each clinical scenario, we offer a treatment algorithm based on our clinical experience. The article also addresses the most common complications associated with SRS and their treatment. PMID- 15527087 TI - Prospective clinical trials of brain tumor therapy: the critical role of neurosurgeons. AB - Prospective clinical trials are critical to the scientific evaluation of new treatments for brain tumors. This paper reviews basic concepts of early and late phase prospective clinical trials that are most relevant to neurosurgical oncologists, with an emphasis on the challenges associated with conducting clinical trials of brain tumor therapies. Novel clinical trial designs that meet these challenges by incorporating pretreatment 'molecular profiling' and post treatment 'molecular endpoints' are described. Because of their ability to obtain brain tumor specimens from patients before and after treatment, neurosurgeons have been required to play an increasingly important role in the execution of these molecular-based clinical trials. Potential avenues for enhancing the participation of neurosurgeons in the design and development of clinical trials are discussed. PMID- 15527086 TI - Pediatric surgical neuro-oncology: current best care practices and strategies. AB - Significant advances have been made in the diagnosis and treatment of childhood brain tumors. Gross total surgical resection combined with appropriate adjuvant therapies can achieve a high rate of disease control for low grade gliomas, ependymomas and medulloblastomas. High grade gliomas, tumors involving the optic apparatus or diencepahalic structures, diffuse brainstem lesions, and recurrent or metastatic disease still pose considerable therapeutic challenges. We review the current treatment strategies of the three most common types of pediatric brain tumors: gliomas, medulloblastomas and ependymomas, and discuss current and future diagnostic and therapeutic modalities. PMID- 15527088 TI - Developing a brain tumor center. AB - The development of a multidisciplinary brain tumor center requires many steps, from selection of leaders to garnering institutional support, growing the clinical neuro-oncology business, and building multidisciplinary involvement especially with the tumor board. Key clinicians and researchers need to be recruited to develop the academic enterprise, and acquiring external endorsement and philanthropy support can be critical factors in maintaining institutional support and in meeting special needs of the center. The process of managing money, space and support personnel, and the maintenance of the overall program are important as well. Our views on building a brain tumor center reflect our specific situation in developing and growing the Hermelin Brain Tumor Center at Henry Ford Hospital in Detroit and hence might not be precisely applicable to the development of centers elsewhere. PMID- 15527089 TI - Skull base surgery for benign skull base tumors. AB - The use of skull base techniques in the treatment of benign skull base tumors has allowed access to areas of the brain once considered inaccessible. The most common benign skull base tumors encountered in neurosurgical practice are benign meningiomas, schwannomas, and glomus jugulare tumors. Gross total resection of these lesions gives patients the best possible chance of a cure. In this paper, we review the rationale for the use of skull base surgery techniques for benign skull base tumors. PMID- 15527090 TI - Evolving role of skullbase surgery for patients with low and high grade malignancies. AB - Malignancy involving the skull base remains a formidable management challenge. Advances and refinements in diagnostic imaging, instrumentation, microvascular reconstruction, and an improved overall appreciation of the anatomy of the skullbase have extended the boundaries of tumor resectability and in some cases, obviated the need for adjuvant therapies. Successful management of high-grade malignancy however, requires a carefully constructed multi-modal treatment plan to maximize patient outcome. Over the course of an 11-year period, 259 patients with skullbase malignancies were treated by the author in the setting of a tertiary care comprehensive cancer center. All patients were evaluated by a multidisciplinary team experienced in the assessment and treatment of skullbase malignancy. Management paradigms were constructed and treatment based on categorization into low or high-grade malignancy was recommended and undertaken. This manuscript discusses this patient population and the outcome of the management paradigms that were constructed. Differences in outcome based on the characterization of malignancies as either low or high grade is discussed. Complications of treatment and patient reported quality of life outcomes are reviewed. PMID- 15527091 TI - The evolving role of stereotactic radiosurgery for patients with skull base tumors. AB - Tumors located at the base of the skull are among the most difficult problems that neurosurgeons encounter. Management of patients with skull base tumors must take into account that complete tumor removal is not possible with acceptable morbidity in many patients. Therefore, radiation therapy and stereotactic radiosurgery are commonly performed. The use of radiosurgery for patients with skull base tumors has increased significantly over the past three decades and we now possess a better understanding of the expectations of skull base radiosurgery. For properly selected patients with benign tumors (meningiomas, schwannomas, glomus tumors), tumor control rates between 90 and 100% have been reported. Local control rates for chordomas and chondrosarcomas have ranged from 50 to 70%, but tumor growth adjacent to the treated area (marginal failure) remains a significant problem. Radiosurgery is also commonly performed for patients with malignant skull base tumors as a palliative treatment and symptom relief is common, especially for patients with facial pain related to their tumor. Follow-up beyond 10 years is still needed to better define long-term results of radiosurgery and the incidence of radiation-induced neoplasms after radiosurgery. PMID- 15527092 TI - Endoscopic and minimally invasive microsurgical approaches for treating brain tumor patients. AB - Recent developments in neuroendoscopy and minimally invasive procedures have greatly impacted the diagnosis and treatment of brain tumors. In this paper, we will review these innovations and discuss how they have influenced our approach to the treatment of intraventricular and pituitary tumors. Finally, the concept of keyhole neurosurgery is illustrated by discussing 'eyebrow orbitotomy' approach as an example. As noninvasive therapeutic alternative become available, future neurosurgeons will be challenged to develop effective and less invasive surgical approaches for the diagnosis and treatment of patients will brain tumors. PMID- 15527093 TI - Surgical strategies for treating patients with pineal region tumors. AB - Optimal management of pineal region tumors depends on securing an accurate histologic diagnosis to facilitate management customized to the nuances of specific pathologies. As an initial step, surgical intervention by either stereotactic biopsy or open surgery is necessary to obtain tissue for pathologic examination. Stereotactic biopsy has the benefit of relative ease and minimal morbidity but is associated with greater likelihood of diagnostic inaccuracy compared to open surgery where more extensive tissue sampling is possible. The role of surgical debulking in the management of pineal tumors is clearly defined for some tumors but is less evident for others. Among the one third of pineal tumors that are benign or low grade, complete surgical resection is achievable and constitutes optimal management with excellent long-term recurrence-free survival. The benefits of aggressive surgical resection among malignant tumors are less clear but several studies have correlated degree of tumor removal with improved outcome. Advances in technology, surgical technique, and post-operative care have minimized surgical complications, however all surgical procedures in the pineal region, including both stereotactic biopsy and open surgery, are potentially hazardous. Advanced judgment, experience, and expertise are necessary to achieve rates of success sufficient to justify aggressive management. Management strategies using stereotactic biopsy, endoscopy, and radiosurgery can also provide favorable outcomes in some cases. Selective incorporation of these innovations can be expected to improve the already highly favorable outcome for all pineal region tumors. PMID- 15527094 TI - Transsphenoidal and transcranial surgery for pituitary adenomas. AB - This paper reviews the progress made over the first century of pituitary surgery. The goals of surgery for pituitary tumors are to eliminate tumor mass effect and perform as complete a removal as possible, retain pituitary function, and normalize any hormonal hypersecretion. Since the initial transsphenoidal approach performed in Austria by Schloffer, the transsphenoidal approach has become the preferred surgical approach to most pituitary tumors. The history and development of the transsphenoidal approach to the sella is discussed, as are the contemporary techniques of microscopic and endoscopic pituitary surgery. The continued evolution of the variations and extension of the transsphenoidal approach to other lesions are reviewed. The indications and use of a transcranial approach to remove pituitary tumors are discussed. More recently, stereotactic radiosurgery (SRS) has become an important adjuvant management technique in the management of difficult pituitary adenomas, especially with cavernous sinus invasion. PMID- 15527095 TI - Stereotactic radiosurgery for pituitary adenomas: a review of the literature. AB - OBJECTIVE: Pituitary adenomas are very common neoplasms and represent between 10 and 20% of all primary brain tumors. Historically, the treatment armamentarium for pituitary adenomas included medical management, microsurgery, and fractionated radiotherapy. More recently, radiosurgery has emerged as a viable treatment option. The goal of this research is to define accurately the efficacy, safety, and role of radiosurgery for treatment of pituitary adenomas. METHODS: Medical literature databases from 1965 to 2003 were searched for articles pertaining to pituitary adenomas and stereotactic radiosurgery. Each study was evaluated for the number of patients, radiosurgical parameters (e.g. tumor margin dose), length of follow-up, tumor growth control rate, complications, and rate of hormonal normalization in the case of functioning adenomas. RESULTS: A total of 34 published studies including 1567 patients were reviewed. Radiosurgery offers a tumor growth control rate of approximately 90%. The reported rates of hormonal normalization for functioning adenomas vary substantially. This range is in part due to widespread differences in endocrinological criteria utilized for post radiosurgical assessment. Thus far, the risks of radiation induced neoplasia and cerebral vasculopathy associated with radiosurgery appear to be lower than for fractionated radiation therapy. The incidence of other serious complications following radiosurgery is quite low. CONCLUSIONS: Although surgical resection typically is the primary treatment modality, stereotactic radiosurgery offers safe and effective treatment for recurrent or residual pituitary adenomas. In rare instances, radiosurgery may be the best initial treatment for patients with pituitary adenomas. Refinements in the radiosurgical technique will likely lead to improved outcomes. PMID- 15527096 TI - Tumors of the osseous spine. AB - Recent advances in molecular biology with the development of novel therapeutic agents, as well as fundamental understanding of the mechanisms of bone metastases have greatly altered the therapeutic options in patients with spine tumors. Improvements in spine instrumentation as well as the development of recombinant Bone Morphogenetic Protein for spine reconstruction and fusion offers promising hope for curative strategies in selected patients. A clearer application of the fundamentals of surgical oncology applied to spine tumors should result in a greater proportion of patients undergoing surgically appropriate en bloc resections for spine tumors. Finally, the superiority of surgery over external irradiation in the controlled clinical trial setting should greatly expand the pool of patients requiring surgery. In this review, we consider the recent advances in primary bone tumors including chordoma, sarcomas, multiple myeloma, as well as metastatic cancer to the spine. PMID- 15527097 TI - Spinal cord and intradural-extraparenchymal spinal tumors: current best care practices and strategies. AB - The management of patients with intradural spinal tumors differs in many respects from approaches taken for patients with intracranial tumors. Intramedullary lesions are often completely surrounded by normal spinal cord, displacing vital functional tracts eccentrically. Extramedullary lesions can drastically compress the spinal cord and nerve roots, reducing normal tissue to a ribbon-like consistency. The small amount of normal tissue relative to tumor has implications for surgery and postoperative adjuvant therapy. In addition, operative intervention must take spinal stability into consideration. In this report, we describe the current best care practices and strategies for patients with a diagnosis of spinal astrocytoma, ependymoma, hemangioblastoma, schwannoma, and meningioma. Treatment of patients with intradural tumors of the spinal cord and adjoining structures has changed over the past 20 years. Advances in many disciplines including neuroradiology, neurosurgery, neurooncology, and neuropathology have contributed to expediting diagnosis and improving outcomes. PMID- 15527098 TI - The evolving role of stereotactic radiosurgery and stereotactic radiation therapy for patients with spine tumors. AB - Traditional management strategies for patients with spinal tumors have undergone considerable changes during the last 15 years. Significant improvements in digital imaging, computer processing, and treatment planning have provided the basis for the application of stereotactic techniques, now the standard of care for intracranial pathology, to spinal pathology. In addition, certain of these improvements have also allowed us to progress from frame-based to frameless systems which now act to accurately assure the delivery of high doses of radiation to a precisely defined target volume while sparing injury to adjacent normal tissues. In this article we will describe the evolution from yesterday's standards for radiation therapy to the current state of the art for the treatment of patients with spinal tumors. This presentation will include a discussion of radiation dosing and toxicity, the overall process of extracranial radiation delivery, and the current state of the art regarding Cyberknife, Novalis, and tomotherapy. Additional discussion relating current research protocols and future directions for the management of benign tumors of the spine will also be presented. PMID- 15527099 TI - Peripheral nerve tumors: management strategies and molecular insights. AB - Because of its relative rarity and a wide variety of clinical manifestations, peripheral nerve tumors (PNTs) often present to specialists from widely different disciplines, thus often resulting in delayed diagnosis and a non-cohesive pattern of management. Critical appraisal of the history and physical examination followed by radiological investigations, by experienced medical personnel, ultimately suggests that the extremity mass is perhaps a PNT, rather than the wide variety and more common soft tissue tumors. Included in this appraisal is a search for a pre-disposition syndrome, the most common of which are neurofibromatosis-1 and -2 (NF1, NF2) and schwannomatosis, which may require life long careful follow up. Intra-operative and post-operative management decisions in a multidisciplinary manner with knowledge of the biological, pathological and clinical behavior of the PNT, is of utmost importance. In the context of pre disposition syndromes, where multiple tumors may exist along with other nervous system lesions, molecular biological insights and hopefully the biological therapies that stem from this knowledge are of interest. In this article the spectrum of PNTs and their management protocols, including the algorithm for treatment of malignant PNTs advocated by our institute, are presented and currently available genetic insights and probable role of experimental therapies are briefly reviewed. PMID- 15527100 TI - Neuro-oncology in a nutshell. PMID- 15527102 TI - Irradiation after surgically induced brain injury in the rat: timing in relation to severity of radiation damage. AB - The aim of this study was to evaluate how timing of irradiation after brain surgery in rats relates to overall extent of brain radiation damage. Extent of injury was determined according to lipid peroxidation (malondialdehyde; MDA) levels in brain tissue. Thirty female rats were randomly assigned to five equal groups (Groups A-E). Four groups underwent trephination and scalpel hemisection of right frontal lobe. Rats in Groups B and D received 25 Gy cranial irradiation in a LINAC system at 10 and 20 days after surgery, respectively. Twenty-four hours later they were killed and their right frontal lobes were removed for lipid peroxidation determination. Groups A and C were not irradiated; these groups were killed and had their frontal lobes removed on day 11 and day 21 post-surgery, respectively. The remaining six animals (Group E, sham surgery) underwent trephination only, and were killed and had their frontal lobes removed 24 h later. There was a significant difference between the mean MDA levels in the control group and Group D, and between the levels in Group B and Group D (P < 0.05 for both). The difference between the mean for Group A and the mean for Group B was even more significant (P < 0.01). The most striking differences were between the control group and Group B, and between Group B and Group C (P < 0.001 for both). The data from this rat model suggest that, in humans, starting radiotherapy early (1-2 weeks) after debulking of a brain tumor may result in significantly higher levels of tissue damage than if the radiation is started 3 weeks or more postoperatively. Further experimental research is needed to project these findings in rats to human subjects. PMID- 15527101 TI - Correlation of N-cadherin expression in high grade gliomas with tissue invasion. AB - Cadherins are Ca2+-dependent cell adhesion molecules that play an important role in tissue construction and morphogenesis in multicellular organisms. Over the last few years, reports have emerged in the literature describing the involvement of cadherins in tumor invasion and metastasis. Cadherins typically demonstrate up and down-regulation according to the biological needs of the tissue. Additionally, up-regulation of N-cadherin is thought to be important for tumor formation in early stages of tumor development. We studied N-cadherin in surgical specimens of patients with primary glioblastoma by microarray analysis and found that N-cadherin mRNA expression is up-regulated compared to normal brain. To study the effects of N-cadherin expression on invasion and metastasis in vitro and in vivo, we overexpressed N-cadherin in the rat C6 glioma cell line which normally has low levels of N-cadherin. We found that up-regulation of N-cadherin resulted in a slight decreased adhesion to type IV collagen, fibronectin, and laminin, but statistically significant decreased adhesion to type I collagen. Furthermore, increased expression of N-cadherin correlated with a dramatic decrease in invasive behavior in extracellular matrix invasion assays. We then proceeded to study these cell lines in vivo in a rat intracranial glioma model, and found that N-cadherin expression inversely correlated with invasion into surrounding tissues, irregular margins, and extracranial invasion. In summary, these data collectively demonstrate that N-cadherin levels are important in the malignant behavior of gliomas, and may serve as a prognostic indicator for patients with high-grade gliomas. PMID- 15527103 TI - Dexamethasone mediated inhibition of local IL-2 immunotherapy is dose dependent in experimental brain tumors. AB - Local delivery of cytokines has been shown to have a potent anti-tumor activity against a wide range of malignant brain tumors. In this study, we examined the role of systemic immunosuppression using dexamethasone on the efficacy of local IL-2 immunotherapy in treating experimental murine CNS tumors. An endothelial cell line secreting hIL-2 (NTC-121) was injected intracranially in C57BL/6 mice (n = 10/ group) along with B16/F10 (wild type) melanoma cells. A separate set of animals also received daily injections of either 1 mg/k or 10 mg/kg of dexamethasone. Sixty percent of mice treated with IL-2 (P < 0.001 vs. control) vs. 55% (P < 0.005) of mice treated with IL-2 and 1 mg/kg of dexamethasone were long-term survivors (LTS) of > 120 days. There was no difference in survival between control animals that received only wild type cells or animals that were treated with IL-2 and 10 mg/kg of dexamethasone. Histopathological examination of brains from animals sacrificed at different times showed an accumulation of CD8 + T-cells around the site of the injected tumor only in the IL-2 group and the group that received 1 mg/kg of dexamethasone. These results suggest that while high doses of dexamethasone can completely inhibit the immune response observed with IL-2, lower and more likely therapeutic doses of dexamethasone do not inhibit local IL-2 immunotherapy. PMID- 15527104 TI - Parent phenotype and age dependence, on rat glioma tumor rejection. AB - BACKGROUND: The brain, despite the blood-brain barrier, does not escape to the highly variable host rejection response mediated by a very strong and complex immune reaction when rat glioma cells are transplanted into the adult animal. METHODS: Crosses were performed among parents that are able or enable to reject a well-known brain tumor cell line (C6). Newborn animals were also challenged with rat glioma cells both in the brain and the side flanks. RESULTS: The percentage of susceptibility or resistance to develop a lethal glioma can be estimated knowing the parental phenotypes. When both parents had rejected an induced tumor, 63% of the progeny will also reject it. Similarly, if both parents died as a consequence of the tumor, 70% of the progeny would also be unable to reject the challenge of glioma C6 cells. Newborn animals do not have a mature immune system and they tolerate transplanted cells much better than adults. We found no rejection to glioma C6, at both brain and side flank sites, in 1-day-old neonatal Wistar rats. Tumors were beginning to be eliminated if the cells are inoculated at day 3 from birth on the flanks, and at 1 week from birth on the brain. CONCLUSIONS: There is a genetic component conferring susceptibility or resistance to the lethal effect of tumor development and progression depending on the parental phenotype of the adult rats. Neonatal rats represent a much more reliable model than adults to study experimental therapies against gliomas. PMID- 15527105 TI - Melanocytoma. PMID- 15527106 TI - Temozolomide for the treatment of recurrent supratentorial glioma: results of a compassionate use program in Belgium. AB - Temozolomide (TMZ) has demonstrated activity and acceptable toxicity for the treatment of recurrent high-grade gliomas in prospective phase II studies. Limited information is available on TMZ when prescribed outside a clinical trial. We conducted a retrospective study to evaluate the activity and safety of TMZ that was prescribed for the treatment of recurrent glioma in the context of a compassionate use program in Belgium. Data were obtained on 117 adult patients (from five hospitals) who received TMZ as first or second line chemotherapy. The recommended starting dose of TMZ was 200 mg/m2 x5d q28d for chemonaive patients and 150 mg/m2 x5d q28d for pre-treated patients. Toxicity was generally mild. Thrombocytopenia was the most frequent treatment related adverse event (grade 3/4 in 17% of patients). Its occurrence was correlated with a starting dose of 200 mg/m2/d and stresses the need to monitor toxicity. The overall objective response rate (complete and partial response) was 29 and 34% of patients achieved an objective disease stabilization. The median progression-free survival was 104 days (95% CI: 85-123) and the median overall survival was 215 days (95% CI: 161 269). In multivariate analysis a 'deep localization' of the glioma (as opposed to a cortico-subcortical localization) and 'the preceding history of a low-grade glioma' were respectively identified as a negative and positive independent prognostic variable for survival. No significant difference in terms of response or median survival was observed between patients with anaplastic astrocytoma or oligo-astrocytoma and chemonaive glioblastoma multiforme. This retrospective study indicates that the reported activity and toxicity profile of TMZ for the treatment of patients with recurrent glioma is reproducible outside the setting of a prospective clinical trial. PMID- 15527107 TI - 123I-IMT SPECT and 1H MR-spectroscopy at 3.0 T in the differential diagnosis of recurrent or residual gliomas: a comparative study. AB - The aim of this investigation was to compare two current non-invasive modalities, single photon emission tomography (SPECT) using 123-iodine-alpha-methyl tyrosine (123I-IMT) and single-voxel proton magnetic resonance spectroscopy (1H-MRS) at 3.0 T, with regard to their ability to differentiate between residual/ recurrent tumors and treatment-related changes in patients pretreated for glioma. The patient population comprised 25 patients in whom recurrent glioma was suspected based on MR imaging. SPECT imaging started 10 min after iv. injection of 300-370 MBq 123I-IMT and was performed using a triple-head system. The IMT uptake was calculated semiquantitatively using regions-of-interest. 1H-MRS was performed at 3.0 T using the single-volume point-resolved spectroscopy (PRESS) technique. Guided by MR imaging volumes-of-interest for spectroscopy were placed into the suspected lesions. Signal intensities of choline-containing compounds (Cho), creatine and phosphocreatine (Cr), and N-acetylaspartate (NAA) were obtained. When using the cut-off of 1.62 for 123I-IMT uptake, the sensitivity, specificity, and accuracy of the 123I-IMT SPECT were 95, 100 and 96%, respectively. For 1H MRS, the sensitivity, specificity and accuracy were 89, 83 and 88%, respectively, based both on the metabolic ratios of Cho/Cr and Cho/NAA as tumor criterion with cut-off values of 1.11 and 1.17, respectively. In conclusion, 123I-IMT SPECT yielded more favorable results compared to 1H-MRS at distinguishing recurrent and/or residual glioma from post-therapeutic changes and may be particularly valuable when the evaluation of tumor extent is necessary. PMID- 15527108 TI - Intracranial meningiomas developing at long intervals following low-dose X-ray irradiation of the head. AB - Five patients are reported who underwent X-ray epilation in childhood for tinea capitis and who developed meningiomas after about four decades. X-ray irradiation resulted in permanent alopecia in four of the five patients. In four patients the tumors were found on the convexity, one patient had a tuberculum sellae meningioma. All five patients underwent surgery. Recurrence was noted in three of the five patients, one of them was reoperated. No malignant features, but signs of atypia were found at histopathological examination. The development of meningiomas after low-dose irradiation with long latency periods, the predominant calvarial location of the tumors, the high recurrence rates, the absence of malignant traits but the presence of atypias are features overwhelmingly common with similar cases published in the literature. The fact that two of the five patients were sisters stresses the importance of genetic factors in the evolution of these tumors. PMID- 15527109 TI - Carcinomatous meningitis as the presenting manifestation of gallbladder carcinoma: case report and review of the literature. AB - The primary tumors that typically cause carcinomatous meningitis include lung cancer, breast cancer, leukemia, lymphoma and melanoma. A variety of neurological signs and symptoms can be seen depending on the extent and location of the meningeal metastasis. Once the diagnosis of carcinomatous meningitis is confirmed, the search for the primary tumor can be a challenge and at times may require extensive radiographic or even surgical evaluation to obtain specimen for pathological confirmation. Here we report a patient who presented with bilateral cranial nerve VIII and cerebellar symptoms, and was diagnosed with carcinomatous meningitis. Only after an exploratory laporatomy did it become clear that the initial symptoms were related to a metastatic gallbladder carcinoma. PMID- 15527110 TI - Intracranial metastatic esthesioneuroblastoma responsive to temozolomide. AB - Successful management of a heavily pretreated 58-year-old woman with metastatic esthesioneuoblastoma using temozolomide is reported. There is no standard treatment of this tumors with extra- and intracranial manifestations. The response, long term stability and high quality of life using temozolomide for this tumor entity should be recognized. PMID- 15527111 TI - Reducing the overall treatment time for radiotherapy of metastatic spinal cord compression (MSCC): 3-year results of a prospective observational multi-center study. AB - BACKGROUND: This prospective multi-center study investigates a reduction of the overall treatment time for radiotherapy of MSCC, which is important for these mostly disabled patients. PATIENTS AND METHODS: Two standard fractionation schedules, 30 Gy/10 fractions/2 weeks (n = 71) and 40 Gy/20 fractions/4 weeks (n = 65) were compared for functional outcome and ambulatory status. Motor function was graded using an 8-point-scale before RT, at the end and at 6, 12 and 24 weeks after RT. A multi-variate analysis was performed for functional outcome. Included variables were the fractionation schedule and the three relevant prognostic factors. These factors are the type of primary tumor, the time of developing motor deficits before RT and the pre-treatment ambulatory status. RESULTS: The ambulatory rates were 49% in the 30 Gy group and 52% in the 40 Gy group before RT (P = 0.888), and 56% and 60% after RT (P = 0.888). Improvement of motor function occurred in 45% of the 30 Gy group and 40% of the 40 Gy group (P = 0.752). The relevant prognostic factors were comparably distributed in both groups. According to the multivariate analysis, a slower development of motor deficits (P < 0.001), a favorable histology (P = 0.040) and being ambulatory (P = 0.045) were associated with better functional outcome, whereas the fractionation schedule had no significant impact (P = 0.311). CONCLUSIONS: The data suggest both schedules to be comparably effective for functional outcome. Thus, 30 Gy/10 fractions/2 weeks should be applied instead of 40 Gy/20 fractions/4 weeks. The reduction of the overall treatment time from 4 to 2 weeks means less discomfort for the paraparetic or paraplegic patient. PMID- 15527112 TI - Testicular seminoma 16 years after treatment for CNS germinoma. AB - Most patients with intracranial germinomas will be cured and become long-term survivors. Physicians caring for these patients should recognize that these patients may be at risk for disease-related and/or treatment-related late sequelae. We report the case of a 27-year-old man who developed testicular seminoma 16 years after treatment for intracranial germinoma. Like their testicular cancer counterparts, long-term survivors of intracranial germinomas may have a susceptibility to develop a subsequent germ cell tumor. These patients require lifelong medical follow-up and should be encouraged to perform testicular self-examination at the appropriate age. PMID- 15527113 TI - Carcinomatous meningitis in cancer of the uterine cervix. AB - Carcinomatous meningitis is extremely rare in cervical cancer. The diagnosis of carcinomatous meningitis is a difficult one when clinical symptoms are limited and radiographic imaging is normal. Demonstration of malignant cells in the cerebrospinal fluid remains the gold standard to establish the diagnosis. For patients without bulky disease who can be treated with radiotherapy, standard treatment for carcinomatous meningitis is chemotherapy, which may be administered intrathecally. Despite the poor prognosis, treatment may result in effective palliation. We describe a 54-year-old patient who was diagnosed with carcinomatous meningitis in the course of metastatic cervical cancer and who responded to administration of intrathecal methotrexate. PMID- 15527115 TI - Ghost peaks in reversed-phase gradient HPLC: a review and update. AB - New examples and sources of ghost peaks in reversed-phase gradient HPLC are described and related to previous publications to give a broad perspective of ghost peak problems. In one new example the ghost peak was found to be due to mixing problems caused by a period of non-delivery of the stronger "solvent B" in a stepped gradient system. In a second example the ghost peak was due to plasticizer contamination of the organic solvent. This paper includes tips and recommendations for the consistent running of ghost-peak-free reversed-phase gradient HPLC. PMID- 15527114 TI - Phase II study of concurrent continuous Temozolomide (TMZ) and Tamoxifen (TMX) for recurrent malignant astrocytic gliomas. AB - PURPOSE AND BACKGROUND: The aim of this study was to assess the frequency of response and toxicity in adults with recurrent anaplastic astrocytoma (AA) or glioblastoma multiforme (GM) treated with concurrent continuous TMZ and TMX. METHODS: In addition to histology, eligibility included age > 18 years, Karnovsky score > or = 60, normal laboratory parameters, no radiotherapy (RT) for 4 weeks, measurable disease and normal EKG. The chief exclusions were: previous TMZ, TMX or dacarbazine (DTIC); nitrosourea within 6 weeks; history of deep venous thrombosis or pulmonary emboli. All patients (pts) had received prior RT. TMZ was given at 75 mg/M2/day for 6 weeks, repeated every 10 weeks, maximum 5 cycles. Four pts received 60 mg/M2/day for 6 weeks due to extensive prior chemotherapy exposure. TMX was started at 40 mg twice daily (b.i.d.) for 1 week and then was increased in three successive weeks to 60, then 80, then 100 mg b.i.d. Response was assessed before every cycle with MRI +/- gadolinium (Gd). RESULTS: Sixteen pts enrolled: GM 10, AA 6; female 6, male 10; median age 48 (21-58); prior chemotherapy 7. There was one partial response and one stable disease. Eleven pts progressed by the end of cycle 1; three pts failed due to toxicity before completing cycle 1. Median time to treatment failure was 10 weeks. The main toxicities were: transaminitis, pancytopenia, 1st division herpes zoster, deep vein thrombosis and fatigue. The study was closed due to the low response rate and frequency of toxicity. PMID- 15527116 TI - Analytical method for the quantitation of sertraline hydrochloride stereoisomers by electrokinetic chromatography. AB - Sertraline is a basic compound and of pharmaceutical application for antidepressant treatment. The compound has two chiral centers. Separation of the three enantiomeric impurities from the parent compound is challenging. In this study, we successfully separated all four stereoisomers by electrokinetic chromatography using highly sulfated gamma-cyclodextrin and highly sulfated alpha cyclodextrin as the chiral selectors. The two chiral selectors provided different selectivity and therefore affected the overall separation profiles. This may be due to the size difference between the dichlorophenyl moiety end and naphthalenamine moiety end, resulting in two different types of inclusion complexes with the different cyclodextrins. For routine analysis, highly sulfated gamma-cyclodextrin was better than highly sulfated alpha-cyclodextrin. For each stereoisomeric impurity, the method using sulfated gamma-cyclodextrin provided a limit of quantitation at or lower than 0.1% of the drug substance with adequate resolution. The critical resolution at this concentration level was not less than 4.0. Experimental data suggested that an internal standard was necessary for the purpose of quantitation, and the practical linearity range for analysis of sertraline stereoisomeric impurities was of about two orders of magnitude. PMID- 15527117 TI - Determination of lycopene in food by on-line SFE coupled to HPLC using a single monolithic column for trapping and separation. AB - A method that would eliminate the degradation of lycopene during analysis was developed. Supercritical fluid extraction (SFE) with carbon dioxide as the extraction medium was connected on-line to high performance liquid chromatography (HPLC) where a single monolithic column was used for trapping and the subsequent separation of analytes. The method was linear over the studied range (0.1-2.5 microg), and it was repeatable (R.S.D. 3.9%), sensitive (LOD = 0.5 ng) and fast (35 min). Lycopene was determined in tomatoes, fruit and several food products. Because of the on-line construction, lycopene was not in contact with air or light during the whole procedure and the amount analysed should therefore correspond to the real amount in the sample. PMID- 15527118 TI - New extraction method for the analysis of linear alkylbenzene sulfonates in marine organisms. Pressurized liquid extraction versus Soxhlet extraction. AB - A new method has been developed for the determination of linear alkylbenzene sulfonates (LAS) from various marine organisms, and compared with Soxhlet extraction. The technique applied includes the use of pressurized liquid extraction (PLE) for the extraction stage, preconcentration of the samples, purification by solid-phase extraction (SPE) and analysis by liquid chromatography with fluorescence detection. The spiked concentrations were added to the samples (wet mass of the organisms: Solea senegalensis and Ruditapes semidecussatus), which were homogenized and agitated continuously for 25 h. The samples were extracted by pressurized hot solvent extraction using two different extraction temperatures (100 and 150 degrees C) and by traditional Soxhlet extraction. The best recoveries were obtained employing pressurized hot solvent extraction at 100 degrees C and varied in the range from 66.1 to 101.3% with a standard deviation of between 2 and 13. Detection limit was between 5 and 15 microg kg(-1) wet mass using HPLC-fluorescence detection. The analytical method developed in this paper has been applied for LAS determination in samples from a Flow-through exposure system with the objective of measuring the bioconcentration of this surfactant. PMID- 15527119 TI - Effect of temperature in reversed phase liquid chromatography. AB - The high temperature liquid chromatography (HTLC) reveals interesting chromatographic properties but even now, it misses some theoretical aspects concerning the influence of high temperature on thermodynamic and kinetic aspects of chromatography: such a knowledge is very essential for method development. In this work, the effect of temperature on solute behavior has been studied using various stationary phases which are representative of the available thermally stable materials present on the market. The thermodynamic properties were evaluated by using different mobile phases: acetonitrile-water, methanol-water and pure water. The obtained results were discussed on the basis of both type of mobile phases and type of stationary phases. Type of mobile phase was found to play an important role on the retention of solutes. The kinetic aspect was studied at various temperatures ranging from ambient temperature to high temperature (typically from about 30 to 200 degrees C) by fitting the experimental data with the Knox equation and it was shown that the efficiency is improved significantly when the temperature is increased. In this paper, we also discussed the problem of temperature control for thermostating columns which may represent a significant source of peak broadening: by taking into account the three main parameters such as heat transfer, pressure drop and band broadening resulting from the preheating tube, suitable rules are set up for a judicious choice of the column internal diameter. PMID- 15527120 TI - Development of a liquid chromatography-electrospray mass spectrometric method for the simultaneous analysis of benzoxazolinones and their degradation products. AB - A new method for the simultaneous analysis of some benzoxazolinones, aminophenoxazinones and malonamic acids was developed based on liquid chromatography (LC) coupled to mass spectrometry (MS), using electrospray ionization (ESI) and operating in positive mode. Different ESI-MS parameters, such as fragmentor voltage, capillary voltage, drying gas flow, nebulizer gas pressure and drying gas temperature, were optimized in order to obtain structural information and to achieve maximum sensitivity. Chromatographic separation was performed by a reversed-phase LC column using a linear gradient of water and methanol. Quality assurance of the developed method was assessed by measuring parameters as linearity, sensitivity, repeatability and reproducibility. Quantification method based on the use of internal standard was developed, selecting synthetic 2-methoxy-2H-1,4-benzoxazin-3(4H)-one as internal standard. Good correlations were obtained for all analytes relative to this compound in the range of 0.05-1.5 ng/microL. Instrumental detection limits were between 0.02 and 0.2 ng/microL. Repeatability and reproducibility studies showed acceptable coefficient of variation values. PMID- 15527121 TI - Multiple headspace solid-phase microextraction of 2-cyclopentyl-cyclopentanone in polyamide 6.6: possibilities and limitations in the headspace analysis of solid hydrogen-bonding matrices. AB - The interactions between a polar analyte, 2-cyclopentyl-cyclopentanone, and a solid polar matrix, polyamide 6.6, during multiple headspace solid-phase microextraction (MHS-SPME) were studied. Strong hydrogen bonding between the analyte and the matrix was observed and shown to cause slow migration and adsorption of the analyte. These matrix effects led to erroneous quantitation despite the use of multiple headspace extraction. Addition of water disrupted the hydrogen bonding between the analyte and the matrix and a valid quantitation was achieved. The addition of water also increased the sensitivity and allowed the identification of 2,5-bis(cyclopentyl)-1-cyclopentanone. The amount of 2 cyclopentyl-cyclopentanone in five different polyamide 6.6 samples was measured using the developed multiple headspace solid-phase microextraction method with water-displacer. The measured concentrations were in the range of 1.44-15.61 microg/g. These concentrations were up to 30% higher than the concentrations measured after microwave-assisted extraction (MAE), which indicates incomplete recovery by MAE. The use of water as a displacer eliminated the matrix effects and complete recovery of the analyte was achieved by MHS-SPME. PMID- 15527122 TI - Gradient elution of organic acids on a beta-cyclodextrin column in the polar organic mode and its application to drug discovery. AB - A high performance liquid chromatographic method was developed that separated organic acids using the polar organic mode. The separation was obtained using a beta-cyclodextrin stationary phase with a mobile phase that was composed of acetonitrile/methanol/triethylamine (TEA)/acetic acid. The compounds were eluted under gradient conditions and the elution order depended on the number, type and position of the hydrogen bonding functional groups present in the molecule. Adjusting the acid to base ratio resulted in the biggest change in selectivity. In addition, increasing the methanol concentration decreased the retention times of the analytes, which had little effect on the selectivity. Using a certain set of conditions one could separate a large number of organic acids, which allowed these acids to be detected by UV and mass spectrometry. These conditions were used to evaluate the purity of potential pharmaceutical drug candidates that showed activity towards a kinase target vascular endothieal growth factor (Vegf). Each compound contained a carboxylic acid group that was critical to the activity. The method was able to give purity estimates of these samples, which were difficult to determine by other HPLC methods. PMID- 15527124 TI - Chitosan as a macroaffinity ligand purification of chitinases by affinity precipitation and aqueous two-phase extractions. AB - (1) Chitosan was found to be a suitable macroaffinity ligand for affinity precipitation of chitinase from Neurospora crassa, cabbage and puffballs. (2) The activity recoveries of 85, 82 and 90% with concomitant fold purifications in terms of specific activities were 27, 15 and 30 with N. crassa, cabbage and puffballs and were obtained with affinity precipitation. These results were obtained with clarified extracts/homogenates as the starting materials. (3) The incorporation of chitosan in poly(ethylene glycol) (PEG)-salt aqueous two-phase system allowed purification of chitinases from these sources directly from unclarified extracts/homogenates. (4) The 96% (w/v) chitosan (of initially introduced into the aqueous two-phase system) partitioned into PEG-phase and this enhanced the partitioning of chitinases into PEG-phase. The chitosan, free as well as bound to chitinases, could be separated from PEG-phase by increasing the pH to 7. (5) By the process of desorption with 2.0 M MgCl2, 86, 80 and 88% activity recoveries (% expressed in terms of total units of enzyme activities in the crude extract) were obtained in the case of N. crassa, cabbage and puffballs, respectively. The corresponding fold purifications in terms of specific activities were 34, 20 and 38. (6) The purified preparations gave single bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the estimated molecular masses agreed with the reported values in the literature. PMID- 15527123 TI - Quantitative determination of dityrosine in milk powders by liquid chromatography coupled to tandem mass spectrometry using isotope dilution. AB - An analytical method to quantify dityrosine (DiTyr) in milk powder samples is presented. The assay is based on isotope dilution liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (LC-ESIMS/MS). The sample preparation entails acid hydrolysis of milk proteins followed by a solid phase extraction (SPE) step. Neither artifactual formation nor degradation of DiTyr were observed during the proteolysis step. Mass spectral detection was performed in the positive ion mode by recording five transition reactions for DiTyr, in order to unambiguously confirm the presence of DiTyr by correct ion ratios. Under the analytical conditions used, the limit of detection (LOD) and limit of quantification (LOQ) for DiTyr were estimated at ca. 2 and 6 micromol DiTyr per mol of Tyr (using ca. 500 microg of milk proteins), with a mean recovery of ca. 90%. Quantification was conduced in eight different commercial milk powder samples, and the level of DiTyr ranged from below the LOQ up to 393.0 +/- 9.1 micromol DiTyr per mol of Tyr. PMID- 15527125 TI - Rapid characterization of oligonucleotides by capillary liquid chromatography nano electrospray quadrupole time-of-flight mass spectrometry. AB - A fast quality control method is developed allowing the desalting and characterization of oligonucleotides by capillary liquid chromatography and on line nano-electrospray ionization quadrupole time-of-flight mass spectrometry using column switching. The influence of addition of ammonium acetate, trans-1,2 diaminocyclohexane-N,N,N',N'-tetraacetic acid, formic acid or acetic acid to the sample, addition of ammonium acetate to the trapping solvent and variation of the trapping time on the further reduction of cation adduction was studied. Final conditions were the addition of 0.1 M ammonium acetate to the sample, the use of a trapping solvent consisting of 0.4 M aqueous 1,1,1,3,3,3-hexafluoro-2-propanol (HFLP) adjusted to pH 7.0 with triethylamine plus 10 mM ammonium acetate during 8 min and the elution of the oligonucleotides with 0.4 M HFIP in 50% methanol. The potential of the optimized procedure is demonstrated for different synthetic oligonucleotides. PMID- 15527126 TI - Chromatographic behavior of pyrithiones. AB - Pyrithione biocides are currently viewed as a major prospect for the replacement of tributyltin antifoulants in ship paints. The chromatographic behavior of 1 hydroxy-2-pyridinethione (pyrithione, PT), bis(2-pyridinyl)disulfide 1,1'-dioxide (PT2), and the metal complexes zinc [Zn(PT)2], iron [Fe(PT)3] and copper [Cu(PT)2] pyrithione, were investigated by means of UV-vis spectroscopy, ESI-MSn, HPLC-DAD and HPLC-ESI-MSnD. This revealed transformations of the analytes, which affect the development of adequate methods for species or environmental analysis of pyrithiones. PT transforms into copper- or iron- containing complexes and/or the oxidation product PT2, depending on the type of the stationary phase used in chromatographic analysis. Speciation complicates direct chromatography of [Zn(PT)2] and [Fe(PT)3]. PMID- 15527127 TI - Simultaneous extraction and fate of linear alkylbenzene sulfonates, coconut diethanol amides, nonylphenol ethoxylates and their degradation products in wastewater treatment plants, receiving coastal waters and sediments in the Catalonian area (NE Spain). AB - The efficiency of six wastewater treatment plants (WWTPs) in the Catalonian region to remove several classes of ionic and non-ionic surfactants was investigated, using solid-phase extraction (SPE) followed by liquid chromatography-mass spectrometry (LC-MS). Occurrence and distribution of linear alkylbenzene sulfonates (LASs), coconut diethanol amides (CDEAs), nonylphenol ethoxylates (NPEOs) and their degradation products were studied in coastal areas receiving WWTP effluents. For the analysis of sediment samples pressurized liquid extraction (PLE) followed by SPE extract clean-up and LC-MS detection was used for simultaneous extraction of all target compounds. Concentrations of parent long ethoxy chain NPEOs in raw water entering WWTPs ranged from 60 to 190 microg/L. In effluents concentrations ranged from 2.8 to 6.6 microg/L, which corresponded to an average primary elimination of 93-96%. Nonylphenol (NP) was found in concentrations from 0.2 to 18 microg/L in WWTP influents and up to 5 microg/L in the treated water, showing a clear declining trend with respect to concentrations reported in the last 5 years. The elimination of LASs and CDEAs varied depending on the WWTP and was higher than 85% in well operated WWTPs with secondary treatment and only 56% (for CDEAs) and 30% (for LAS) in WWTPs with primary treatment. Levels up to 86 mg/kg of LAS, 1.7 mg/kg of NP, 400-760 microg/L for NPEOs and NPECs were detected in sediments at sampling points near the discharge of WWTP effluents, clearly showing that in coastal area receiving WWTP effluents surfactants and their degradation products are widespread contaminants. PMID- 15527128 TI - New considerations about the separation and quantification of antimony species by ion chromatography-hydride generation atomic fluorescence spectrometry. AB - A new method for the speciation of inorganic [Sb(III) and Sb(V)] and organic (Me3SbCl2) antimony species by using a polystyrene-divinylbenzene-based anion exchange HPLC column (Hamilton PRP-X100) coupled to hydride generation atomic fluorescence spectrometry (HG-AFS) is presented. Several mobile phases were tested for the baseline separation of these three antimony species, investigating in detail experimental parameters such as concentration and pH. The best efficiency and resolution was achieved by using a gradient elution between diammonium tartrate 250 mmol l(-1) pH 5.5 (A) and KOH 20 mmol l(-1) pH 12 (B). The gradient programme used was 100% B for 1.5 min, decreasing to 0% B in 0.1 min and maintained the elution with 100% A for 5.5 min. Analysis time was less than 7 min. Equilibration of the column with the complexing mobile phase was found to be critical in order to avoid Sb(III) double peak formation. Dilution in diammonium tartrate medium was necessary in order to avoid Sb(III) oxidation at microg l(-1) concentration level. Detection limits of 0.06 microg l(-1) for Sb(V), 0.09 microg l(-1) for Me3SbCl2 and 0.04 microg l(-1) for Sb(III) as well as repeatability and reproducibility better than 5% R.S.D. (n = 10) and 9% R.S.D. (n = 30) (for 1 and 5 microg l(-1) of Sb(V) and Sb(III) and 5 and 10 microg l(-1) of Me3SbCl2) were obtained. Accuracy and recovery studies were carried out by analysing one river freshwater sample and two water certified reference materials. The proposed methodology can be considered reliable and straightforward for antimony speciation in fresh water samples. PMID- 15527129 TI - Separation and determination of lanthanides, thorium and uranium using a dual gradient in reversed-phase liquid chromatography. AB - Separation and determination of lanthanides, Th and U is of great relevance in different fields of science and technology. Reversed-phase high-performance liquid chromatography (RP-HPLC) using alpha-hydroxy isobutyric acid (alpha-HIBA) as an eluent on reversed-phase column modified to cation exchanger has been reported to achieve the separation. However, under those conditions, Th and U are eluted amongst lanthanides, making their quantification difficult due to overlapping with some of the lanthanides peaks. In this work, different chromatographic parameters (concentrations of eluent and ion interaction reagent, pH, etc.) were studied systematically to arrive at optimum chromatographic conditions. Using the dual (concentration and pH) gradient conditions, lanthanides, Th and U could be separated in 11 min by RP-HPLC with sequential elution of Th and U after the elution of all the lanthanides. The separation methodology was tested using SY-3 rock sample for the separation and determination of lanthanides, Th and U. The method allows an accurate determination of these elements in a single run using a single column. Also, the method is fast and cost-effective compared to the reported methods. PMID- 15527130 TI - Combining headspace gas chromatography with mass spectrometry detection for confirmation of hydrocarbon residues in virgin olive oil following automatic screening. AB - Hydrocarbon residues can be present in virgin olive oils either naturally or as contaminants. Presence of benzene, toluene, ethylbenzene, xylene isomers and styrene (BTEXS) in virgin olive oil, demanded the establishment of a cut-off level to discriminate oil samples containing these residues at normal levels from those at high levels caused by contamination. By introducing volatile components present in the headspace (HS) of the oil samples (without prior chromatographic separation) into the ionization source of a mass spectrometer, samples were classified as containing normal or high levels of BTEXS (recommendable or non recommendable forhuman consumption). Confirmation and quantification of contaminated virgin olive oils were achieved by combining HS sampling with gas chromatography-mass spectrometry (GC-MS). No sample pretreatment was necessary for the chromatographic method, but the addition of 600 microl of ethyl acetate as chemical modifier for 10 ml of virgin olive oil. Detection limits ranged between 3 and 9 ng/ml. Oil analysis showed the presence of toluene in all samples, as well as the absence of benzene, ethylbenzene and o-xylene. PMID- 15527131 TI - Determination of chloroanisole compounds in red wine by headspace solid-phase microextraction and gas chromatography-mass spectrometry. AB - The objectives of this study, were the development and validation of an analytical method for the determination of 2,4,6-trichloroanisole (TCA), 2,3,4,6 tetrachloroanisole (TeCA) and pentachloroanisole (PCA) in red wine by headspace solid-phase microextraction and GC-MS as well as the application of the optimized and validated method for the quatification of chloroanisoles in different red wines from Navarra. To carry out this study, the extraction variables have been optimized. The fiber and the experimental design selected permit the determination of low analyte concentrations (ng/L) with good accuracy (<5%). Moreover, an analytical method for the determination of TCA and TeCA in wine by GC-MS has been validated. The results obtained in the validation step, recovery values, detection and quantitative limits, and precision were acceptable for all the analytes in the ranges of concentration studied (<5% and <10% for TCA and TeCA, respectively). This method has been used as an analytical method for the quantification of TCA and TeCA in red wine samples that were selected for this study, yielding good results. PMID- 15527132 TI - Correlation between emitted and total amount of 2-cyclopentyl-cyclopentanone in polyamide 6.6 allows rapid assessment by HS and HS-SPME under non-equilibrium conditions. AB - A correlation was found between the emitted and total amount of 2-cyclopentyl cyclopentanone in polyamide 6.6. The emitted amounts were measured by GC-MS after headspace (HS) or headspace solid-phase microextraction (HS-SPME) and the total content was determined after microwave-assisted extraction (MAE). The correlation was valid also under non-equilibrium conditions, which allows rapid assessment of 2-cyclopentyl-cyclopentanone content in polyamide 6.6 by headspace techniques. The incubation time needed for non-equilibrium headspace analysis could be reduced from 5 h to 45 min if the PA66 granules were milled to powder prior to extraction. However, to reach equilibrium between the analyte in the solid sample and the headspace still required 12 h of incubation at 80 degrees C. The long incubation time is explained by slow diffusion rate due to the strong hydrogen bonding between analyte and matrix and the relatively high crystallinity of polyamide 6.6. The headspace extraction profile showed several equilibrium-like patterns that are easily mistaken for the real equilibrium. PMID- 15527133 TI - Increasing UV detection sensitivity in the supercritical fluid chromatographic analysis of alcohol polyethers. AB - Alcohol ethoxylates (AEOs) that contain a wide distribution of oligomers pose a challenge for ultraviolet (UV) absorbance detection due to the fact that the AEOs absorb strongly only in the range of commercial UV detectors between 190 and 200 nm. Most mobile phase components, with the exceptions of water and carbon dioxide, also absorb in this region. Ethoxylated hexadecanol and octadecanol were derivatized with disilazane-chlorosilane mixtures for the formation of phenyl containing silylethers. Derivatized samples were analyzed by supercritical fluid chromatography (SFC) coupled with both electrospray ionization mass spectrometry and UV absorbance detection. An increase in the number of phenyl groups incorporated into the derivatives increased the number of oligomers observed by UV detection. An increase in the number of oligomers detected increased the calculated average molar ethoxylate values. The average molar oligomer values calculated by SFC-UV for these alcohols were consistent with the nominal reported values. PMID- 15527134 TI - Biocatalyzed acidolysis of soybean oil triacylglycerols to increase oleic acid content. AB - Lipase catalyzed acidolysis of triacylglycerols (TAG) of soybean oil with oleic acid in organic solvent was studied; immobilized lipase from Rhizomucor miehei was used and the effects of reaction time, incubation temperature and enzyme load on TAG total and positional fatty acid (FA) percentage compositions were investigated. The results show that oleic acid incorporation was high after 24 and 48 h, while after 72 h a lower level of oleic acid in TAG was observed. Moreover, for the reactions carried out at 30 and 40 degrees C, it was observed that the oleic acid level was about 46.5% while ligtly higher values (about 49%) were observed at 50 degrees C; however, under this last condition, the modification of sn-2 position FA composition was higher. Finally, the variable enzyme load resulted also important on the incorporation of oleic acid; in particular, even if the value of 10% (w/w) of enzyme load resulted in a lightly lower incorporation of oleic acid in soybean oil TAG (about 45%), in this situation a minimal modification of sn-2 position FA composition was obtained. PMID- 15527135 TI - Micellar electrokinetic chromatography with bile salts for predicting ecotoxicity of aromatic compounds. AB - The retention factors of several aromatic compounds were obtained by micellar electrokinetic chromatography (MEKC) using cholate, taurocholate, deoxycholate and deoxytaurocholate as micellar systems. The possibility of using these retention factors to describe and predict several ecotoxicological activities of different aromatic compounds was evaluated. Adequate correlations retention ecotoxicity (log LC50 in fish and daphnia, log EC50 in green algae and daphnia, chronic values in fish and green algae, bioconcentration factor, and soil sorption coefficient) were obtained for the micellar systems studied. The predictive ability of the models obtained for these micellar systems was compared. Predicted values concur with the experimental log LC50 in Bluegill, Rainbow trout, Fathead minnows and Daphnia Magna values for the compounds studied. The results obtained indicated the usefulness of the MEKC systems investigated for the rapid ecotoxicity assessment of aromatic compounds. PMID- 15527136 TI - Retention behaviour of peptides in capillary electrochromatography using an embedded ammonium in dodecacyl stationary phase. AB - The potential of a silica stationary phase bearing an embedded cationic quaternary amine in dodecacyl chain, to separate peptides by capillary electrochromatography (CEC) has been evaluated. The ability of this stationary phase, to generate a consistent anodic electroosmotic flow was first evaluated. This flow was found to be independent of pH over a wide range (2-12), of the acetonitrile percentage in the electrolyte. The stability of the stationary phase evaluated through the electroosmotic flow variations was demonstrated at extreme pH values (2.5 and 9.1). A careful examination of the influence of mobile phase conditions (acetonitrile percentage, salt concentration and nature of buffer) on the electrochromatographic retention and electrophoretic migration behaviour of different standard peptides was carried out. In acidic conditions, the electrokinetic contribution appears to be predominant compared to the chromatographic one. Several types of chromatographic interactions, reversed phase partitioning and anion exchange, were involved in the CEC of peptides, whereas repulsive electrostatic interaction could be considered as negligible. This stationary phase affords different selectivity compared to that observed on a C18 stationary phase. Finally, the method was applied to the peptide mapping of beta-lactoglobulin and human growth hormone under unpressurized and isocratic elution. PMID- 15527137 TI - Analysis of aged red wine pigments by capillary zone electrophoresis. AB - Red wines of different ages (1-14 years) were analysed by Capillary zone electrophoresis (CZE). Pigments were separated using a fused silica capillary with an effective length of 56 cm and an internal diameter of 75 cm. Disodium tetraborate at 50 mM with a pH of 9.4 was used as buffer solution with methanol as modifier. Electrophoregrams were recorded at 280, 520 and 599 nm and spectra were measured from 200 to 599 nm using a diode array detector. Several differences in the CZE signals obtained for different aged wines were observed, as well as between red and white wines. The signals of a group of seven peaks increased in mature wines. A decrease in free anthocyanins was also identified in aged wine. The migration times of these compounds corresponded to charge/size ratios higher than those of free anthocyanins, and their spectra suggested combinations of anthocyanins with flavanols or tannins. The global CZE response of these pigmented polyphenolic polymers was linearly correlated with the spectrophotometric determination of polymeric pigments (96%) and age (92%). PMID- 15527138 TI - Insights into the retention mechanism of neutral organic compounds on polar chemically bonded stationary phases in reversed-phase liquid chromatography. AB - The solvation parameter model is used to characterize the retention properties of a 3-aminopropylsiloxane-bonded (Alltima amino), three 3-cyanopropylsiloxane bonded (Ultrasphere CN, Ultremex-CN and Zorbax SB-CN), a spacer bonded propanediol (LiChrospher DIOL) and a multifunctional macrocyclic glycopeptide (Chirobiotic T) silica-based stationary phases with mobile phases containing 10 and 20% (v/v) methanol-water. The low retention on the polar chemically bonded stationary phases compared with alkylsiloxane-bonded silica stationary phases arises from the higher cohesion of the polar chemically bonded phases and an unfavorable phase ratio. The solvated polar chemically bonded stationary phases are considerably more hydrogen-bond acidic and dipolar/polarizable than solvated alkylsiloxane-bonded silica stationary phases. Selectivity differences are not as great among the polar chemically bonded stationary phases as they are between the polar chemically bonded phases and alkylsiloxane-bonded silica stationary phases. PMID- 15527139 TI - Preparation and characterization of long methacrylate monolithic column for capillary liquid chromatography. AB - Long methacrylate monolithic columns (100 cm x 320 microm i.d.) were prepared from silanized fused-silica capillaries of 320 microm i.d. by in situ copolymerization of butyl methacrylate (BMA) with ethylene dimethacrylate (EDMA) in the presence of a suitable porogen. The separation performance and selectivity of the column were evaluated and compared with a 25 cm x 320 microm i.d. column prepared in the same way by capillary high-performance liquid chromatography (micro-HPLC). The results showed that the 1 m long monolithic column can generate 33 x 10(3) plate number and exhibited good permeability, higher sample loadability, and separation capability. PMID- 15527140 TI - Rapid liquid chromatography-ultraviolet determination of organic acids and phenolic compounds in red wine and must. AB - A reversed-phase liquid chromatography-ultraviolet (LC-UV) method is proposed for the rapid simultaneous analysis of the main carboxylic acids and polyphenols in must and wine. Good resolution was obtained for citric, tartaric, malic, lactic, acetic, caffeic, ellagic and gallic acids, (-)-epicatechin, quercetin and resveratrol. A novel silica-based column containing ether-linked phenyl groups, with polar end-capping and suitable for low-pH aqueous mobile phases was used and found to be superior to others tested. The method employed a mixture of 0.2% TFA in water and acetonitrile as eluents, showed linearity and precision, and was applied to samples of must and wine. PMID- 15527141 TI - Preparative isolation and purification of hydroxyanthraquinones and cinnamic acid from the chinese medicinal herb Rheum officinale Baill. by high-speed counter current chromatography. AB - A high-speed counter-current chromatography (HSCCC) method for preparative separation and purification of five hydroxyanthraquinones and cinnamic acid from the Chinese medicinal herb Rheum officinale Baill. was developed by using pH gradient elution. The purities of rhein, emodin, aloe-emodin, chrysophanol, physcion and cinnamic acid were all over 98%, as determined by high performance liquid chromatography (HPLC). The structures of them were identified by 1H NMR. PMID- 15527142 TI - Preparative isolation and purification of coumarins from Angelica dahurica (Fisch. ex Hoffn) Benth, et Hook. f (Chinese traditional medicinal herb) by high speed counter-current chromatography. AB - A preparative high-speed counter-current chromatography (HSCCC) method for isolation and purification of coumarins from Angelica dahurica (Fisch. ex Hoffin) Benth, et Hook. f(Baizhi in Chinese) was successfully established by using n hexane-methanol-water as the two-phase solvent system in gradient elution mode. The upper phase of n-hexane-methanol-water (5:5:5, v/v) was used as the stationary phase of HSCCC. The mobile phase used in HSCCC was the lower phase of n-hexane-methanol-water (5:5:5, v/v) and n-hexane-methanol-water (5:7:3, v/v) that was changed in gradient. Three major components including imperatorin, isoimperatorin and oxypeucedanine were isolated, each at over 98% purity as determined by high-performance liquid chromatography (HPLC). The peak fractions of HSCCC were identified by 1H-NMR and 13C-NMR. PMID- 15527143 TI - Preparative isolation and purification of gastrodin from the Chinese medicinal plant Gastrodia elata by high-speed counter-current chromatography. AB - Gastrodia elata Blume is a famous Chinese medicinal plant, which has been widely used for the treatment of rheumatism, epilepsy, paralysis, hemiplegia, lumbago, headache and vertigo. High-speed counter-current chromatography was successfully used for the first time for the preparative isolation and purification of the bioactive component gastrodin from G. elata Blume. The crude gastrodin was obtained by extraction with ethanol from the dried roots of G. elata Blume under sonication. Preparative high-speed counter-current chromatography with a two phase solvent system composed of n-butanol-ethyl acetate-water (2:3:5, v/v/v) was successfully performed yielding 48 mg gastrodin at 96% purity from 500 mg of the crude extract (10.3% gastrodin) with the recovery of approximately 90% in a one step separation. PMID- 15527144 TI - Comparison of three derivatization ways in the separation of phenol and hydroquinone from water samples. AB - Hydroquinone and phenol were analysed in aqueous alkaline model samples using chromatographic techniques. The compounds were isolated by solid-phase extraction and determined quantitatively by capillary gas chromatography after derivatization. The effectivity of derivatization was tested in three phases of sample handling: (i) before extraction; (ii) in the extraction bed; and (iii) during elution. PMID- 15527145 TI - Chemical composition of the essential oil of Salvia multicaulis Vahl. var. simplicifolia Boiss. growing wild in Lebanon. AB - The essential oil of aerial parts of Salvia multicaulis Vahl. var. simplicifolia Boiss. (Lamiaceae) growing wild in Lebanon was obtained by hydrodistillation and was analysed by GC and GC-MS. 67 compounds constituting 95.2% of the oil were identified, the major components being alpha-copaene (8.0%), alpha-pinene (6.6%), myrtenol (5.7%), trans-sabinyl acetate (5.3%). PMID- 15527146 TI - Global tobacco control must become a top public health priority. PMID- 15527147 TI - Current challenges in tobacco control. AB - Tobacco is the world's biggest preventable killer, but the circumstances of its history, the power and influence of its commerce and the nature of addiction make it a very difficult public health issue. Determinants of smoking are both individual and environmental. Genetics and environment influence to varying degrees all of the steps in a smoker's career. Persistence of use, degree of addiction to nicotine and difficulty in stopping are influenced by inherited traits and nicotine susceptibility, whereas the social environment and the individual's cognitions are the key factors in starting smoking and successfully stopping smoking. The tools available to tobacco control include influencing the social and cultural norms concerning tobacco; legislative and regulatory measures to protect the population and to limit tobacco industry marketing tactics, now encapsulated in the Framework Convention on Tobacco Control; and programmes to enhance the chance of not starting and successfully stopping. Strategies for tobacco control must work at both societal and individual levels, and directions are being taken that include genetic, pharmacological, behavioural, socio cultural and international approaches. PMID- 15527148 TI - Pentaglobin in steroid-resistant severe acute respiratory syndrome. AB - SETTING: The treatment of severe acute respiratory syndrome (SARS) is at best controversial, although there is considerable anecdotal experience to show the benefits of corticosteroid therapy for selected patients. Some patients deteriorate relentlessly despite treatment with antibiotic, corticosteroid and mechanical ventilation. OBJECTIVE: To attempt to determine the clinical efficacy of pentaglobin, an IgM-enriched immunoglobulin preparation, on 12 severe SARS patients who continued to deteriorate despite corticosteroid and ribavirin therapy. DESIGN: Retrospective analysis of daily quantitative and radiographic data on the cohort in a regional teaching hospital. RESULTS AND CONCLUSION: There was significant improvement in radiographic scores, when compared with day 1, on days 5, 6 and 7 (P < 0.05) after commencement of pentaglobin treatment. Similarly, there was significant improvement in oxygen requirement, when compared with day 1, on days 6 and 7 (P < 0.05) after commencement of pentaglobin treatment. There were no reported adverse events attributable to pentaglobin administration. Ten patients made an uneventful recovery after treatment. One elderly man died from cardiorespiratory arrest despite clinical and radiological improvement, and another patient is making good progress. Pentaglobin is safe and probably effective in the treatment of steroid-resistant SARS. A double-blind placebo-controlled study should therefore be considered. PMID- 15527149 TI - Bronchial hyperresponsiveness in two populations with different prevalences of atopy. AB - SETTING: Random population samples of young adults from Tartu, Estonia (n = 307) and Uppsala, Sweden (n = 498) in the framework of the European Community Respiratory Health Survey (ECRHS). OBJECTIVE: To compare the prevalence and risk factors for bronchial hyperresponsiveness (BHR) to methacholine in two centres with similar climate and ethnicity but differences in the prevalence of atopy, asthma and respiratory symptoms. DESIGN: General population-based cross-sectional survey. RESULTS: The prevalence of BHR using the cut-off points 1.5, 1.0 and 0.5 mg was 19%, 12% and 8% in Tartu and 11%, 7% and 2%, respectively, in Uppsala. Current smoking was a common risk factor for BHR in both centres. Female sex (odds ratio [OR] 2.9), sensitisation to cat (OR 5.9) and visible mould in the home (OR 2.4) were independent risk factors for BHR in Tartu. In Uppsala, BHR was significantly associated with total IgE levels (OR 2.0) and exposure to environmental tobacco smoke (OR 3.3). CONCLUSION: The high prevalence of BHR can explain the high prevalence of respiratory symptoms in Tartu and indicates that causes other than asthma and atopy can be responsible for the high prevalence of BHR in a population. PMID- 15527150 TI - Non-tuberculous mycobacteria: patterns of isolation. A multi-country retrospective survey. AB - OBJECTIVE: To collect data on non-tuberculous mycobacteria (NTM) isolated from clinical laboratories in different countries to establish: 1) whether the isolation of NTM was increasing, 2) which species were increasing, and 3) whether there was any pattern of geographical distribution. DESIGN: In 1996, the Working Group of the Bacteriology and Immunology Section of the International Union Against Tuberculosis and Lung Disease contacted 50 laboratories in different countries for the necessary information. RESULTS: The number of patients reported with NTM was 36099 from 14 countries. Mycobacterium avium complex, M. gordonae, M. xenopi, M. kansasii and M. fortuitum were the five species most frequently isolated. There was a significant upward trend for M. avium complex and M. xenopi. Pigmented mycobacteria predominated in Belgium, the Czech Republic and the Mediterranean coast of Spain. Non-chromogenic mycobacteria were found to be predominant in the area of the Atlantic coast of Brazil and in Turkey, the United Kingdom, Finland and Denmark. CONCLUSIONS: There was an increase in the number of NTM isolated from clinical samples of patients. Isolation of the most frequent species is constantly changing in most of the geographical areas, and newer species are emerging due to better diagnostic techniques to detect and identify NTM. PMID- 15527151 TI - Characterization of drug-resistant isolates of Mycobacterium tuberculosis derived from Russian inmates. AB - SETTING: Tuberculosis ward of a prison in Russia. OBJECTIVE: Molecular characterization of drug-resistant isolates. DESIGN: Isolates were collected from all tuberculosis patients occurring in the prison over a 1-year period. RESULTS: Of 130 patients studied, 17 patients produced pan-susceptible isolates and 113 produced isolates resistant to at least one drug, including 85 multidrug resistant isolates. Mutations at katG315 occurred in 98% of isoniazid-resistant isolates. Mutations in rpoB were found in 89% of rifampicin-resistant isolates. Mutations in pncA occurred in 13% of the 75 isolates tested. By spoligotyping, members of the Beijing (55 isolates) and LAM (31 isolates) families were identified. By IS6110 genotyping, two groups (34 and 55 isolates) of related isolates were found, including three clusters (10, 12, and 16 isolates) with identical patterns. In a study of samples collected 3 months apart from 28 patients, four patients produced isolates containing a mixture of strains and five patients produced specimens containing distinctly different isolates. Isolates of nine patients acquired additional drug resistance. CONCLUSION: Three families of strains accounted for much of the drug-resistant tuberculosis in this population. Multiple resistance, acquisition of resistance, and infection with two or more strains as well as reinfection were observed. PMID- 15527152 TI - Impact of the human immunodeficiency virus on the epidemiology of tuberculosis in area 15 of the Valencian community in Spain. AB - SETTING: Area 15 in Valencia. OBJECTIVES: To describe the epidemiology (1987 2001) of tuberculosis (TB) in human immunodeficiency virus (HIV) patients. METHODS: Study of annual incidence, age distribution, excess cases attributed to HIV, etiological risk fraction (ERF), population attributable fraction (PAF) and f factor. RESULTS: Of 476 cases diagnosed, 459 were TB, 16 environmental and one mixed; 76% of environmental cases were HIV-positive (P < 0.001). There was a mean annual TB incidence of 24.6/100000, with an annual reduction of 4%. Seventy-three patients were HIV coinfected (16%) (mean incidence 3834/100 000 seropositives). The principal risk factor was drug use (59%) for HIV+ and contact with TB for HIV . We found no difference in pulmonary or extra-pulmonary location between groups, contrary to mixed cases (P < 0.001). In HIV+ there was a lower frequency of infiltrates (P < 0.001) and cavitation (P < 0.01), and a higher frequency of adenitis (P < 0.001), miliary or nodular pattern and normal X-ray (P < 0.001). Seropositives had a 174 times higher probability of developing TB. The mean ERF attributed to HIV was 99%, the PAF was 16% and the f factor was 1.19. Highly active antiretroviral therapy (HAART) reduced the risk of TB in HIV+ by 80%. CONCLUSIONS: TB has continued its decline, although HIV generated an excess of cases in the 1990s. HAART has reduced the TB risk in HIV+ and possibly the overall rate of TB. PMID- 15527153 TI - Transmission characteristics of tuberculosis in the foreign-born and the Canadian born populations of Alberta, Canada. AB - SETTING: All notified cases of tuberculosis in the province of Alberta, Canada, 1994-1998. OBJECTIVE: To compare the transmission characteristics of tuberculosis among foreign-born and Canadian-born cases. DESIGN: Retrospective analysis using DNA fingerprinting (IS6110 restriction fragment length polymorphism and spoligotyping) and patient information from the Alberta Tuberculosis Registry. Transmission indexes were determined by calculating the average number of culture positive pulmonary cases generated by a single source case. RESULTS: Of the 750 cases of active tuberculosis, 437 (58.3%) were in the foreign-born. DNA fingerprinting of Mycobacterium tuberculosis isolates from all 573 culture positive cases over the 5 years from 1994 to 1998 showed that there was significantly less clustering among foreign-born isolates (9.8%) compared to Canadian-born non-Aboriginal (28.8%) and Aboriginal (44.7%) isolates. The transmission index was significantly higher for males, lower for those > or =65 years of age, and higher for Aboriginals. CONCLUSION: Although cases of tuberculosis in the foreign-born constitute the majority in Alberta, there is little transmission to other foreign-born or to Canadian-born individuals. Transmission of tuberculosis among the Aboriginal population remains a significant problem in Alberta. PMID- 15527154 TI - Functional and genetic assessment of IFN-gamma receptor in patients with clinical tuberculosis. AB - OBJECTIVE: The molecular basis of the genetic vulnerability underlying the most common form of clinical tuberculosis (TB) remains largely unknown. We speculated that mild genetic defects in the interferon-gamma (IFN-gamma) signalling pathway caused a subtle functional impairment of IFN-gamma which would explain susceptibility to Mycobacterium tuberculosis in clinical TB. DESIGN: A case control study. RESULTS: We evaluated functional responsiveness to IFN-gamma in monocytes from patients with clinical TB (n = 10), and analysed the genetic sequences of the IFN-gamma receptor 1 (IFN-gammaR1) and STAT1 genes in patients with disseminated TB (n = 18). IFN-gamma stimulated an increase in the expression of HLA-DR and CD64 on monocytes of both controls and patients; the rate of increase in expression was the same in both groups. Treatment with IFN-gamma before lipopolysaccharide (LPS) stimulation further increased tumour necrosis factor-alpha (TNF-alpha) production as compared to TNF-alpha production with LPS stimulation alone; the rate of increase in TNF-alpha production was the same in both groups. The known mutations in the coding sequences of the IFN-gammaR1 and STAT1 genes were not found in the patients with disseminated tuberculosis. CONCLUSION: These results suggest that impairment of the IFN-gamma signalling pathway did not account for cases of clinical TB in this study. PMID- 15527155 TI - Tuberculin skin testing in BCG-vaccinated populations of adults and children at high risk for tuberculosis in Taiwan. AB - SETTING: Various populations at high risk for tuberculosis (TB) infection and with high bacille Calmette-Guerin (BCG) vaccination coverage in eastern Taiwan. OBJECTIVE: To investigate the usefulness of the tuberculin skin test (TST) to diagnose TB in BCG-vaccinated populations. DESIGN: Cross-sectional survey. RESULTS: TST reaction size was recorded for 444 individuals ranging in age from 8 months to 99 years, of whom 94.3% had scars to suggest previous BCG vaccination. The TST-positive (> or =10 mm) rate was increased in all comparisons of higher to lower TB risk, including household contacts (relative risk [RR] 3.52, P < 0.0001) and intermediate risk residents (RR 2.30, P = 0.030) compared to a lower risk control group. Advancing age was generally associated with increases in the TST positive rate. Gender or the number of BCG vaccinations had no relation to the TST-positive rate. CONCLUSIONS: The results of this survey suggest that in eastern Taiwan a positive TST represents either active or latent TB infection rather than past BCG vaccination. Therefore, high BCG vaccination coverage in this region does not appear to limit the usefulness of the TST as a tool for diagnosing TB. PMID- 15527156 TI - Quality assessment of smear microscopy for acid-fast bacilli in the Argentine tuberculosis laboratory network, 1983-2001. AB - SETTING: Tuberculosis Laboratory Network, Argentina. OBJECTIVES: 1) To evaluate the technical quality of smear microscopy for acid-fast bacilli (AFB) supervised in 1983-2001, and 2) to analyse the effect of procedural errors on the results. DESIGN: Registers of technical evaluation results for AFB microscopy were analysed. The quality of specimens, smears, staining and readings were evaluated, as was the relationship between these parameters. RESULTS: The proportion of good quality specimens was considered acceptable. A direct relationship was demonstrated between positivity in results and mucopurulent sputum. The proportion of thin smears was relatively high. Positivity and bacillary count were lower in thin smears. Staining quality was considered good. The average agreement in readings throughout the country was 98%. Nevertheless, the false positive rate was considered significant, and 46% of false-positive results were associated with defective staining. CONCLUSIONS: The technical quality and agreement in the laboratory network were satisfactory. Nevertheless, improvements need to be made in the following: the quality of the smears, staining and reading, coverage, decentralisation of supervision, the slide selection method and data registration. Operational research on the storage and preservation of slides is also necessary. PMID- 15527157 TI - Outcome of tuberculosis retreatment in routine conditions in Cotonou, Benin. AB - SETTING: National Tuberculosis Programme (NTP), Cotonou, Benin. OBJECTIVE: To study the patient characteristics and outcome of tuberculosis retreatment cases in a well-functioning NTP. METHODS: A retrospective, register-based study of all smear-positive pulmonary tuberculosis cases put on retreatment (2SERHZ/1ERHZ/5R3H3E3) between 1992 and 2001 in Cotonou. For comparison, information on new smear-positive cases in Cotonou in 1999 was entered and analysed. RESULTS: Of 8103 tuberculosis patients registered, 642 were put on retreatment. The analysis is mainly based on the 236 patients whose initial treatment regimen records were available (113 relapses, 84 failures, 39 returns after default). Most of the relapse (57%) and return after default (72%) cases were put on retreatment within 12 months after stopping their initial treatment. Overall, the retreatment results were satisfactory (78% success) and comparable with those of new cases (82%); the failure rates were low (3%), as were those for initial treatment (1%). There were more defaulters from retreatment among those who had already defaulted from initial treatment (21%). Treatment success rates were better among women than men. CONCLUSION: The standardised retreatment regimen is effective in Cotonou, probably because the NTP is functioning well, there are no drug shortages, drug taking is strictly supervised, and a good treatment plan is followed. PMID- 15527158 TI - Effectiveness of community-based directly observed treatment for tuberculosis in an urban setting in Tanzania: a randomised controlled trial. AB - SETTING: An urban district in Dar es Salaam city with a high tuberculosis (TB) caseload. OBJECTIVE: To evaluate the effectiveness of community-based direct observation of treatment (DOT) using guardians and former TB patients compared to hospital-based DOT in an urban setting in Tanzania. DESIGN: Unblinded randomised control trial conducted in five sites under operational conditions in Temeke district. No changes to existing treatment delivery were made other than randomisation. The main outcome measure was treatment success. Analysis was by intention to treat. FINDINGS: A total of 587 new tuberculosis patients were enrolled. Among enrolled patients, 260 were assigned to community-based DOT using guardians and former TB patients and 327 to health facility-based DOT. Both DOT options gave similar treatment outcomes. Treatment success rate among patients under community and health facility-based DOT were 85% and 83%, respectively (OR 1.17, 95%CI 0.75-1.83). CONCLUSION: Community-based DOT is as effective as health facility-based DOT and can achieve good treatment outcomes, even in countries with well functioning National Tuberculosis Programmes. PMID- 15527159 TI - A survey of tuberculosis hospitals in India. AB - SETTING: Hospitals with beds for tuberculosis (TB) in India. OBJECTIVES: To assess diagnostic and treatment practices at institutions offering secondary or tertiary level care for TB patients, and to determine the resources being used at these institutions. DESIGN: Countrywide cross-sectional survey of TB hospitals using a mailed semi-structured questionnaire sent to all 105 hospitals with 100 or more beds and to all State Directorate Health Services. RESULTS: The 94 hospitals that returned the questionnaire had 15773 TB beds, one third of the total TB beds in the country. Nearly 1 million patients sought treatment in the TB hospitals and one third were diagnosed with TB; the ratio of smear-positive to smear-negative patients was 1:2.7. Sixty-four per cent of hospitals prescribed unobserved rifampicin in the continuation phase, and 56% of sputum smear-positive patients were hospitalised. The annual expenditure for the TB hospitals was more than the total annual budget for the TB control programme of the country. CONCLUSIONS: In view of the high number of patients seen and the suboptimal practices observed, urgent steps should be taken to ensure implementation of correct diagnostic and treatment policies in hospitals with TB beds. PMID- 15527160 TI - Acceptability of traditional healers as directly observed treatment providers in tuberculosis control in a tribal area of Andhra Pradesh, India. AB - SETTING: Vizianagaram district of Andhra Pradesh, India, where 9% of the population is tribal. OBJECTIVE: To 1) examine traditional healers' (THs) diagnostic and treatment practices for tuberculosis (TB); 2) understand health seeking behaviour for TB amongst tribal groups; 3) evaluate the acceptability of THs to public health workers; and 4) assess how THs can collaborate with the TB Programme. METHODS: In-depth interviews with 120 THs, 37 exit interviews and four focus group discussions with TB patients; interviews with eight non-governmental organisations (NGOs) and 38 public health staff. RESULTS: There were 18 THs per 1000 population. Of the 120 THs interviewed, 15 (13%) claimed to be TB specialists. Of the 72 healers aware of TB, 65 (90%) considered prolonged cough an important symptom and 53 (74%) believed that TB could be cured with allopathic drugs. THs felt collaboration was possible by directly observed treatment (DOT) providers (49, 68%), referring symptomatic individuals (54, 75%) and offering treatment for side effects (18, 25%). CONCLUSION: Involvement of THs in the TB Control Programme may improve services as they are a major health service provider in tribal areas and are acceptable as DOT providers to patients, public health providers and NGO workers. PMID- 15527161 TI - Traditional healers participate in tuberculosis control in The Gambia. AB - SETTING: Twenty-three Gambian villages. OBJECTIVE: To evaluate the feasibility of involving traditional healers in tuberculosis diagnosis and treatment in The Gambia. DESIGN: Twenty-eight traditional healers were educated in the recognition of signs and symptoms of tuberculosis and indications for referral. They administered medications to confirmed cases, and were revisited after 1 year to assess knowledge retention. RESULTS: Over 6 months, the traditional healers referred 66 suspects, from whom eight cases were diagnosed. All were successfully treated. Twenty-three of 24 traditional healers re-interviewed retained appropriate knowledge; 16 continued to refer suspects. CONCLUSIONS: Traditional healers can play a positive role in tuberculosis control. PMID- 15527162 TI - Measurement in epidemiology. AB - Data (information) are the building blocks of epidemiological research. Careful measurement, recording and handling of information are crucial to the research process. The nature of the data required must be precisely defined and its collection requires the use of reliable tools or instruments. Standardisation of definitions and procedures used in measurement helps to ensure comparability. Care in undertaking measurements and rigorous training of personnel carrying out the measurements help to ensure precision. Special care is necessary to ensure completeness and accuracy in recording the data. Information may be obtained from routine or especially designed forms and evaluated by counting deaths or cases of disease or measures of dysfunction. Questionnaires are a frequently used instrument for collecting information. They should include data identifying the individual being studied, followed by specific questions concerning diseases and determinants. To be useful, such questionnaires must be carefully structured. Diseases are usually compared by calculating 'rates' or 'risks', consisting of numerators (cases, events) and denominators (population). Prevalence and incidence are standard measures of 'risk'. The collection of high quality information (data) and its use to construct estimates of 'risk' or risk ratios form the basis of epidemiology. PMID- 15527163 TI - A case of pulmonary tuberculosis associated with IgA nephropathy. PMID- 15527164 TI - Pulmonary alveolar microlithiasis with epididymal and periurethral calcifications causing obstructive azospermia. PMID- 15527165 TI - Epidemiology and control of tuberculosis in Western European cities. PMID- 15527166 TI - Murine model for non-IgE-mediated asthma. AB - There is increasing evidence that inflammatory mechanisms other than atopy or eosinophilic inflammation may be involved in the pathogenesis of asthma. The mechanisms associated with non-atopic (non-IgE) or neutrophil-mediated asthma are poorly investigated. Non-atopic airway inflammation and hyperresponsiveness was induced in mice by skin sensitization with dinitrofluorobenzene (DNFB) followed by intra-airway challenge with dinitrobenzene sulfonic acid (DNS). Acute bronchoconstriction and mast cell activation were observed shortly after challenge. Increased levels of the major mast cell mediator, TNF-alpha, were found in the bronchoalveolar lavage fluid of DNFB-sensitized. Mast cells play a key role in the early release of TNF-alpha since mast-cell-deficient WBB6F1-W/Wv mice did not show an increase in TNF-alpha release after DNFB-sensitization and DNS challenge compared to their ++ littermates. Features of the late-phase pulmonary reaction included mononuclear and neutrophilic cell infiltration, pulmonary edema, in vitro tracheal hyperreactivity and in vivo airway hyperresponsiveness. These characteristics bear marked similarity with those observed in non-atopic asthmatic patients. Therefore, this model can be used to further study the mechanisms potentially responsible for the development of non IgE-mediated asthma. PMID- 15527167 TI - Inflammation induced by inoculation of the joint with Candida albicans. AB - In humans Candida albicans is the most frequently isolated opportunistic fungal pathogen. In immunocompromized host the balance with the commensal fungus easily turns to life-threatening disseminated infection. The asymptomatic Candida persistence in organs and the recurrent infections suggest continuous circulation of yeast cells and their degradation products. Under certain conditions, joints might become one of the infectious sites. More easily a reactivation and destructive process can be provoked in individuals with established arthritis. We have investigated the joint inflammation caused by inoculation of the paw with live C. albicans, in intact mice and mice with collagen-induced arthritis (CIA). The results demonstrate that C. albicans infection when localized into the joints caused rapidly progressing septic arthritis. The effect was associated with a strong swelling, a rapid influx of polymorphonuclear (PMN) cells, and an elevated secretion of TNF-alpha and IFN-gamma by lymph node cells. Joint infection exacerbated the established CIA which correlated with an increased level of anti collagen antibodies. PMID- 15527168 TI - Aberrant [correction of Abberant] cytosolic calcium ion mobilization in chronic granulomatous disease neutrophils. AB - Beside the inability to produce superoxide ions, neutrophils (PMN) from chronic granulomatous disease (CGD) patients show other functional defects, e.g. abnormal membrane potential reactions. We observed that PMN from a female CGD carrier, with a discrete mutation in one allele of the pg91(phox) gene and exhibiting extreme lyonization, showed a consistently and remarkably delayed PMN cytosolic calcium response to the tripeptide fMLP or leukotriene B4 (LTB4). In keeping with results from other CGD patients, membrane potential changes were abnormal, whereas chemotaxis and adherence was normal. Since phospholipase D-generated metabolites are important for calcium transients we examined the generation of phosphatidic acid, but found that to be normal. A male CGD patient with pg91(phox) deficiency exhibited a trend toward prolongation of this calcium response, whereas two other CGD patients (one with p47 and one with 67(phox) deficiencies) had normal calcium transients. Thus, our finding points to a defect of the stimulus response coupling for fMLP and LTB4, which is supposed to be characteristic for this patient or a subset of CGD patients. PMID- 15527169 TI - IL-4 and IL-10 inhibition of spontaneous monocyte apoptosis is associated with Flip upregulation. AB - Human peripheral blood monocytes undergo spontaneous apoptosis in culture. Spontaneous monocyte apoptosis is regulated by the death ligand, Fas Ligand (FasL) binding to its receptor Fas. The pro-inflammatory molecules, LPS and IL 1beta, prevent spontaneous monocyte apoptosis. Here, we demonstrate that the anti inflammatory cytokines IL-4 and IL-10 inhibit spontaneous monocyte apoptosis compared to control-treated cells. IL-4- or IL-10-mediated suppression of spontaneous monocyte apoptosis is associated with the induction of Flip, an essential inhibitor of the Fas-death signal. In contrast, IL-4 and IL-10 inhibit LPS or IL-1beta induced pro-inflammatory cytokine production. These data suggest that in monocytes IL-4 or IL-10 has a dual function, to inhibit pro-inflammatory cytokine production and to suppress spontaneous apoptosis. PMID- 15527170 TI - Lipopolysaccharide protection from oxygen toxicity: effect on rat pulmonary selectins. AB - Sublethal doses of LPS result in increased tolerance to high concentrations of oxygen and this is associated with decreased pulmonary inflammation in a rat model. To investigate the mechanism of decreased neutrophil influx into the lung in this model, we measured levels of mRNA in the lung for the endothelial adhesion molecules, E-selectin and P-selectin. Immunostaining for E-selectin protein was also done in rat lungs, as well as measurement of soluble L-selectin in the blood. These levels were measured in the lungs of adult rats injected with 0.5 mg/kg LPS or placebo at 0 and 24 h and exposed to > 95% O2 for 60 h. Oxygen exposure resulted in significant increases in both E- and P-selectin mRNA and in E-selectin protein after 60 h. LPS resulted in an early rise in E-selectin protein followed by a decline to less than control (saline/O2) levels at 60 h. Messenger RNA for E-selectin followed a similar trend, although there were no differences at 60 h between LPS and control groups exposed to O2. P-selectin mRNA expression did not significantly differ between LPS and control O2 groups. Soluble L-selectin levels decreased by 6 h after LPS infusion and were significantly lower than saline/O2 controls through 24 h, suggesting binding to endothelium. In conclusion, the decrease in E-selectin expression on the surface of pulmonary endothelium after LPS could contribute to decreased inflammation in this model of oxygen toxicity. Soluble L-selectin may serve a further anti inflammatory role after LPS infusion by binding to pulmonary endothelium. PMID- 15527171 TI - Impact of site-specific nucleobase deletions on the arthritogenicity of DNA. AB - Oligodeoxynucleotides (ODN) containing unmethylated CpG motifs (CpG ODN) potently stimulate the innate and acquired immune system. We have compared the in vivo and in vitro inflammatogenic properties of CpG ODNs containing a specific nucleobase deletion either 5'-upstream (ODN-2) or 3'-downstream (ODN-3) of the CpG motif, comparing with a prototype CpG ODN (ODN-1). The frequency of arthritis was similar after intra-articular (i.a.) injections of ODN-1 or ODN-3, but was significantly lower (p < 0.02) after i.a. injections of ODN-2. In vitro production of the pro-inflammatory cytokine TNF-alpha was higher in mouse spleen cell cultures exposed to ODN-2 in comparison to ODN-1. In addition, the level of IL-10 induced by ODN-2 was higher than that induced by ODN-1. On the other hand, TNF-alpha, IL-10, and MCP-1 levels, as well as splenocyte proliferative responses were all significantly lower for ODN-3 than for ODN-1. These results suggest that a 5'-upstream nucleobase deletion reduces arthritogenicity, while maintaining or increasing the production of pro- and anti-inflammatory factors. In contrast, a 3'-downstream nucleobase deletion has no effect on arthritogenicity, despite significantly lower levels of proliferation and pro- and anti-inflammatory cytokines, compared with ODN-1. This study indicates that specific structural elements within the ODN sequence but outside the CpG motif, modulate the immunostimulatory properties of CpG ODNs. PMID- 15527172 TI - An in vitro investigation of the anti-inflammatory properties of potassium humate. AB - In this study the anti-inflammatory potential of potassium humate, derived from bituminous coal, has been investigated in vitro. Exposure of resting and phorbol 12-myristate-13-acetate (PMA) stimulated human neutrophils to potassium humate resulted in a decreased expression of CR3 by activated, but not resting cells, in a dose-related way. Humate also inhibited the adhesion of PMA-stimulated neutrophils to a baby hamster kidney cell line expressing ICAM1 (the CR3 ligand) (BHK331-7). Similar results were obtained using normal BHK cells indicating that this inhibition does not only target specific adhesion molecules on the neutrophil and eosinophil membrane by activated phagocytes, but also affects other mechanisms involved in cell adhesion. Opsonised Sephadex or FMLP/Cyto B induced degranulation of neutrophils and eosinophils were also decreased by humate treatment. Inhibition of the adhesion of activated phagocytes, as well as inhibition of the release of granule polypeptides, both of which are responsible for tissue damage during inflammatory processes, are attractive targets for anti inflammatory drugs. Because humate is well tolerated with an excellent safety profile it merits further evaluation in patients suffering from inflammatory conditions. PMID- 15527173 TI - [80% less myocardial infarct risk through lifestyle changes]. PMID- 15527174 TI - [What is a cost-benefit analysis?]. PMID- 15527175 TI - [Migraine in women. Relationship to menstrual cycle and treatment concepts]. PMID- 15527176 TI - [The "andropause"--fact or fiction?]. PMID- 15527177 TI - [Optimal and future capable]. PMID- 15527178 TI - [Local anesthetics and pain therapy in pharyngitis]. PMID- 15527180 TI - [Hair loss-what can one do?]. PMID- 15527179 TI - [Treatment of patients with intestinal cancer. Increased neurotoxicity after oxaliplatin and related liver metastasis]. PMID- 15527181 TI - Wither academic forensic science? PMID- 15527182 TI - Quantitative analysis of amplifiable DNA in tissues exposed to various environments using competitive PCR assays. AB - Competitive PCR assays were established for the mitochondrial DNA hypervariable region I and the human amelogenin locus. Using these assays, the copy numbers of DNA participating in PCR (amplifiable DNA) were quantified in tissues exposed to different environments. Human ribs, skin and nails were left in three exposure conditions (in the open air, in soil and in water). The amounts of amplifiable DNA in these tissues were quantified during a time period of up to two months. The amount of amplifiable DNA was well preserved in hard tissues (ribs and nails) regardless of the exposure conditions, whereas the soft tissues immersed in water showed a rapid decrease in amplifiable DNA. Strong PCR inhibition was observed in the DNA extracts obtained from buried bones. This phenomenon was clearly identified from an amplification failure of the internal standards in the competitive PCR. A preliminary examination to identify the PCR inhibitor suggested that the soil itself contributed to the inhibition. In addition, the amounts of amplifiable DNA in case samples were also investigated. PMID- 15527183 TI - Textile damage caused by vapour cloud explosions. AB - The aim of the project was to investigate the damage to garments caused by particular vapour cloud explosions. The authors would like to be able to provide investigators with specific information on how to link clothes to a specific type of crime: a particular case study was the inspiration for the examinations. Experiments were carried out in the fire reconstruction chamber of the laboratory using a selection of 26 clothes and 15 household garments differing in colour, fibre composition and textile construction. PMID- 15527184 TI - Measuring the impact of forensic science in detecting burglary and autocrime offences. AB - As UK investment in forensic science has increased, the government has taken a fresh interest in how far this has led to dividends in terms of the detection of crime and its reduction. The Home Office funded 'Pathfinder Project' sought to monitor and document the complex relationships between the collection and use of forensic material (looking at a range of forensic science techniques) and its impact on crime detection. The project specifically targeted the 'volume' crimes of burglary and vehicle crime. Detailed data was gathered on all stages of the process between the collection and use of forensic material and crime detection. The model falls into two conceptual phases--scene attendance to suspect identification and identification to detection. From the analysis it was found that approximately one third of burglary and autocrime scenes are visited by SOCOs. While scientific identifications are only made in a minority of burglary and autocrime offences overall, it belies their importance. About one in ten of burglary and autocrime cases are cleared up by the police and it is estimated that fingerprints and SGMPlus were a contributory factor in achieving one third of these clear ups. PMID- 15527185 TI - Volatile organic compounds from the combustion of human and animal tissue. AB - The volatile by-products of the combustion of ordinary fuels such as wood, polystyrene, polyethylene, urethane foam, PVC and the like are well known to the forensic fire debris examiner. When a fire involves a human body, volatile species are produced that are not so well known, including various alkenes and aldehydes. These have sometimes been mistaken for the residues of unusual accelerants. In an attempt to document what volatiles are produced by the combustion of animal fat and human fat, the authors have used an open-tube pyrolysis probe as a microfurnace to burn small samples of unembalmed subcutaneous fat from human, avian and porcine sources, and collect volatiles by charcoal strip adsorption. The volatile products were analyzed by GC/MS. Predominant species included aldehydes in the C6-C10 range, homologous series of alkenes and alkanes, and other hydrocarbon products. These results were compared to those obtained by free-burning (open flame in air) of similar specimens and to the volatiles detected in debris from beneath a human cadaver in a test fire. Differences between the volatile profiles produced by human fat as compared to pork and chicken fat and adventitious sources of such volatiles are discussed. PMID- 15527186 TI - [Should acupuncture be reimbursed? Three acupuncture programmes of German Statutory Health Insurance Funds (SHI) and their scientific evaluation]. AB - An estimated 40,000 physicians offer acupuncture treatment to patients in Germany. Due to a decision of the German Federal Committee of SHI-accredited Physicians and Health Insurance Funds, acupuncture treatment for chronic low back pain, headache and osteoarthritic pain may be reimbursed from 2000 onwards if patients and physicians participate in specific scientifically evaluated model projects of the Statutory Health Insurance Funds. Currently, three separate acupuncture programmes are offered by different Statutory Health Insurance Funds. The scientific evaluation of all these programmes includes both randomised trials (comparing acupuncture to no treatment, sham acupuncture or standard treatment in 300 to several thousands of patients) and large cohort studies. Details of the three programmes will be presented in the following chapters. PMID- 15527187 TI - [German acupuncture studies]. PMID- 15527188 TI - [Program for evaluation of patient care with acupuncture--the scientific comparison of models and results]. PMID- 15527189 TI - [Design of models for acupuncture in disease and in disease models]. PMID- 15527190 TI - [Assessment of unconventional medical therapies by the Medical Review Board of the Statutory Health Insurance Funds (MDK)]. AB - The Medical Review Board of the Statutory Health Insurance Funds (MDK) provides assessments of medical diagnostic and therapeutic procedures comprising the assessment of both individual cases and systematic assessments of health technologies (HTA). Here the foundation of these assessments will be outlined and demonstrated that assessment work is based on the principles of evidence-based medicine. There are no clear-cut criteria distinguishing between so-called complementary and other medical procedures. It is argued that there should be no difference in the assessment standards and criteria to be applied. This position seems to be supported by a broad consensus that also includes proponents of complementary medicine. PMID- 15527191 TI - [Is there any leeway for assessment beyond randomised clinical trials?]. AB - Randomised controlled trials provide the basis for assessing treatment efficacy. Yet decisions concerning the introduction of new medications and treatment regimens will have to consider more aspects than the ones that are usually assessed in trials. Here, two examples will be discussed to explain the options that health care systems provide. PMID- 15527192 TI - [Evidence-based medicine and public health law: statutory health insurance]. AB - Beyond all differences in terminology and legal principles between the laws governing private health insurance, the governmental financial support for civil, servants and statutory health insurance the fundamental issues to be solved by the courts in case of litigation are quite similar. But only a part of these refer to the quality of medical services, which is the main concern of Evidence based Medicine (EbM); EbM, though, is not able to contribute towards answering the equally important question of how to distinguish between "treatment" and "(health-relevant) lifestyle". The respective definitions that have been developed in the particular fields of law are only seemingly divergent from each other and basically unsuitable to aid the physician in his clinical decision making because the common blanket clauses of public health law are regularly interpreted as rules for the exclusion of certain claims and not as a confirmatory paraphrase of what is clinically necessary. If on the other hand medical quality is what lies at the core of litigation, reference to EbM may become necessary. In fact, it is already common practice in the statutory health insurance system that decision-making processes in the Federal Committee being responsible for quality assurance (Bundesausschuss) are based on EbM principles and that in exceptional cases only the courts have to medically review the Federal Committee's decisions. PMID- 15527193 TI - [The current jurisdiction of the Federal Administrative Court concerning the public service employer's obligation within the scope of the Public Support Fund for Civil Servants]. AB - Civil servants are entitled to a subsidy for their health insurance expenses provided the medical interventions are basically necessary and the amount of related expenses adequate. The definition of the borderline between accepted and unaccepted expenses for medical treatment is factually based on the criteria of evidence-based medicine. Accordingly, expenses are not refundable if incurred for treatments and medical products whose efficiency is doubtful or which generally lack scientific acceptance, though under narrowly defined conditions, even "outsider methods" might be approved of. Thus, the law governing these subsidies is more closely related to private insurance than to social security law. PMID- 15527194 TI - [CAMbase--the realisation of an XML-based bibliographical database system for complementary and alternative medicine]. AB - The term "Complementary and Alternative Medicine" (CAM) describes a variety of approaches to medical theory and practice that have not been commonly accepted by representatives of conventional medicine. This is why research results from this field of medicine are seldom reported in well-established medical journals. Due to the high level of interest and demand for complementary medicine, though, there is an urgent need for a system covering the scientific literature of complementary medicine. Against this background, the chair of Medical Theory and Complementary Medicine at the University of Witten/Herdecke has launched the CAMbase project. The database can be accessed online and is free of charge or additional costs at www.cambase.de. METHODS: Using XML-based protocols, existing databases of Complementary and Alternative Medicine were imported and indexed in a central database for online retrieval. XML-based interfaces, which have been developed within this project (implementation of Open Archives Initiative protocols), can be used for embedding CAMbase into larger bibliographical networks. RESULTS: Since the relaunch of CAMbase in August 2003 an average of 107 search queries per day have been registered. An initial analysis of the search terms has demonstrated that CAMbase is not only used by scientists and researchers but also by both health professionals in clinical practice and patients in search for information about complementary therapies. PERSPECTIVES: In the long term the accessibility of full-text versions of original publications on Complementary Medicine will have to be improved. The further development of CAMbase in co-operation with the Library of Witten/Herdecke University is currently being planned. PMID- 15527195 TI - [Medical errors and iatrogenic injury--results of 173 Schlichtungsstellen proceedings in general practice]. AB - The Schlichtungsstelle (expert panel for alternative dispute resolution) of Northern Germany receives and decides on large numbers of malpractice claims. We report on 173 panel decisions on claims involving general practitioners: Medical negligence was found in 40%, and patient-injuries due to negligence in 28% of these cases. Treatment-caused injuries of the patient were also found in several of the cases decided against the claimant. In proceedings on the grounds of injections, 26 of 30 claimants concerned had suffered iatrogenic injuries. Treatment-caused injuries were severe in 40 of the 173 patients, fatal in 7. Injury rated and margins of error are much lower in industry than in medicine. Reviews of hospital records identified adverse events in 3-4% of hospitalized patients. Autopsy studies revealed that in 10-50% of cases the diagnosis verified postmortem had been missed clinically. Effective risk-management should be based on a trustful relationship among the persons working together: so that errors and adverse events might be discussed openly and the roles of persons, organization or system can be laid open. PMID- 15527196 TI - [Triangulation or how different research methods may lead to different results]. AB - A study of general practitioners' attitudes towards rehabilitation demonstrates how a mix of methods (focus group, case analysis, survey) does not only provide confirmation and validation of research results but also reveals divergences and contradictions. On the one hand, these differences will uncover the weaknesses and strengths of the different methods used but they will also indicate the existence of parallel realities. Different ways to interpret and integrate such divergences will be presented. Triangulation will help to avoid premature conclusions and formulate the conclusions in a such a way that special consideration will be given to the complexity of the subject under investigation. PMID- 15527197 TI - [Oxygen treatment. 1904]. PMID- 15527198 TI - Effects of Qi-therapy on premenstrual syndrome. AB - This study investigated the effects of Qi-therapy (nine sessions over two menstrual cycles) on pain and other symptoms in women with premenstrual syndrome (PMS). Forty-six women who were attending college were randomly allocated to receive Qi-therapy (Qi-therapy group: n = 23) or placed on a waiting list as controls (n = 22: 1 dropped out). Qi-therapy had a significant effect on pain and water retention. In addition, there were significant short-term effects on pain, mental depression, and anxiety. These results suggested that Qi-therapy might be useful as a nursing intervention for women who suffer from PMS to maintain or restore a balance of Qi. PMID- 15527199 TI - Effects of multifunctional fabrics on cardiac autonomic tone and psychological state. AB - Heart rate variability was compared in 20 subjects wearing multifunctionalfabric (experimental sessions) and cotton (control sessions) clothing. Anxiety, depression, fatigue, and stress levels were lower and emotional levels were higher during the experimental sessions than in the control sessions. Multifunctional fabrics reduced the low-frequency/high-frequency power ratio of heart rate variability. These results support the hypothesis that multifunctional fabrics increase cardiac parasympathetic tone. In addition, subjects had lower heart rates during the experimental sessions compared with controls, suggesting a stabilizing effect on the autonomic nervous system. In conclusion, multifunctional fabrics may act to stabilize both the autonomic nervous system and psychological state. PMID- 15527200 TI - Prenatal maternal biochemistry predicts neonatal biochemistry. AB - Depressed (n = 45) and nondepressed (n = 47) mothers were recruited prenatally at an ultrasound clinic. Their urine samples were assayed for cortisol, catecholamines (norepinephrine, epinephrine, dopamine) and serotonin. Their urines were assayed again at the neonatal period, and their newborns' urines were also assayed at that time. The depressed versus the nondepressed mothers showed significantly higher cortisol and norepinephrine and significantly lower dopamine levels across the pre- and postnatal assessments. At the postnatal assessment all levels had decreased except the serotonin levels for both groups. Regression analyses on the mother's postnatal biochemistry with the prenatal biochemistry entered as predictor variables showed highly significant, specific relationships between each of the catecholamines, cortisol, and serotonin. The newborn's biochemistry (except for epinephrine) was higher than the maternal biochemistry. Regression analyses on the neonatal biochemistry with the mother's prenatal biochemistry entered as predictor variables also suggested highly significant, specific relationships. The continuity between the mother's and the newborn's neurotransmitter/ neurohormone profiles and data showing that elevated norepinephrine and cortisol predict to low birthweight and prematurity, respectively, highlight the importance of assessing these levels during pregnancy. PMID- 15527201 TI - Fixation of the mandible changes masseter muscle activity associated with swallowing. AB - The purpose of this study was to investigate the effect of mandibular fixation on activity of the swallowing-related muscles during swallowing. Electromyograms in the masseter muscle, orbicularis oris superior muscle, suprahyoid muscles, and infrahyoid muscles were recorded from seven healthy humans. Electromyographic activity during swallowing was compared between an experimental condition with mandibular fixation and an experimental condition without such fixation. Duration of the swallowing phase was prolonged with mandibular fixation. Mandibular fixation significantly delayed the onset of masseter muscle activity during swallowing, although no other muscle groups showed a delay. The amount of muscle activity during swallowing was not significantly different between the two experimental conditions in any of the muscle groups. In conclusion, mandibular fixation decreases the masseter muscle activity at the initial phase of swallowing. This may be due to decreased need of the masseter muscle activity against force pulling the mandible. PMID- 15527202 TI - ATP-sensitive potassium channels and endogenous adenosine are involved in spinal antinociception produced by locus coeruleus stimulation. AB - The effects of locus coeruleus stimulation on nociceptive evoked discharges of thalamic parafascicular (PF) neurons were investigated in lightly urethane anesthetized rats, aiming to study the mechanisms underlying these effects. Intrathecal (i.t.) administration of aminophylline (an adenosine antagonist), glibenclamide (an ATP-sensitive potassium [K+(ATP)] channels blocker), nicrorandil (Nico; an agonist of K+(ATP) channel and a K+(ATP) channel opener), and 5'-N-ethylcarboxamido-adenosine (NECA; an adenosine agonist) were used. The results showed that (1) locus coeruleus stimulation significantly inhibited the nociceptive evoked discharges of parafascicular neurons, (2) locus coeruleus stimulation-produced antinociception in PF neurons was blocked by both it. glibenclamide and i.t. aminophylline, (3) nociceptive discharges of PF neurons were also suppressed by both i.t. NECA and i.t. nicorandil, and (4) i.t. glibenclamide showed no effect on the suppression of nociceptive discharges induced by NECA, whereas aminophylline blocked the suppression of nociceptive discharges induced by nicorandil. These results suggest that (a) K+(ATP) channels and endogenous adenosine may be involved in the mediation of antinociception induced by norepinephrine, which is released in the dorsal horn by descending fibers originating from the locus coeruleus and (b) the opening of K+(ATP) channels may precede the release of endogenous adenosine in the process of suppressing nociceptive transmission at the spinal level. PMID- 15527204 TI - Opioid neurotoxicity: comparison of morphine and tramadol in an experimental rat model. AB - Histopathologic changes in rat brain due to chronic use of morphine and/or tramadol in progressively increased doses were investigated in this study. Thirty male Wistar rats (180-220 g) were included and divided into three groups. Normal saline (1 ml/kg) was given intraperitoneally as placebo in the control group (n = 10). Morphine group (n = 10) received morphine intraperitoneally at a dose of 4 mg/kg/day for the first 10 days, 8 mg/kg/day between 11-20 days, and 12 mg/kg/day between 21-30 days. The tramadol group (n = 10) received the drug intraperitoneally at doses of 20, 40, and 80 mg/kg/day in the first, second, and the third 10 days of the study, respectively. All rats were decapitated on the 30th day and the brain was removed intact for histology. The presence and the number of red neurons, which are a histologic marker of apoptosis, were investigated in the parietal, frontal, temporal, occipital, entorhinal, pyriform, and hippocampal CA1, CA2, CA3 regions. Red neurons were found in morphine and tramadol groups but not in the control group. The total number of red neurons was not different in morphine and tramadol groups, but the numbers of red neurons were significantly higher in the temporal and occipital regions in tramadol group as compared with the morphine group (p < .05). In conclusion, chronic use of morphine and/or tramadol in increasing doses is found to cause red neuron degeneration in the rat brain, which probably contributes to cerebral dysfunction. These findings should be taken into consideration when chrome use of opioids is indicated. PMID- 15527203 TI - Simultaneous recording of eeg and direct current (DC) potential makes it possible to assess functional and metabolic state of nervous tissue. AB - It has been proposed to assess functional and metabolic state of the brain nervous tissue in terms of bioelectrical parameters. Simultaneous recording of the DC potential level and total slow electrical activity of the nervous tissue was performed in the object of study by nonpolarizable Ag/AgCl electrodes with a DC amplifier. The functional and metabolic state of the brain was determined in terms of enhancement or reduction in the total slow electrical activity and positive or negative shifts in the DC potential level. PMID- 15527205 TI - Electrophysiological and quantitative electroencephalographic measurements after treatment by transcerebral magnetic fields generated by compact disc through a computer sound card: the Shakti treatment. AB - Quantitative electroencephalographic activity over the left and right frontal, temporal, parietal, and occipital lobes was obtained on 4 successive weeks in 12 subjects before they were exposed cerebrally for 30 min to one of two configurations of weak complex magnetic fields or to a sham-field condition. The two configurations were Shakti (c Todd Murphy) and the set of 4 solenoids (Koren boxes) generating a burst-firing magnetic field through the temporal lobes. Compared to baseline measurements there were no statistically significant differences in treatments for relative changes in power over these regions within the delta, theta, low alpha, beta, or gamma ranges. However within the high alpha range (10.5 Hz to 13 Hz), there was a significant interaction between session and type of treatment that was due primarily to the Shakti treatment. These changes were congruent with the subjective experiences reported by some users of the technology. PMID- 15527206 TI - Visual evoked potential is superior to triple dose magnetic resonance imaging in the diagnosis of optic nerve involvement. AB - The aim of this study was to evaluate whether VEP is sensitive to optic neuritis (ON) when compared with triple dose orbital MRI. Twenty-four relapsing-remitting MS (RRMS) patients were included in the study. Group I (n = 10) patients with acute ON, Group II (n = 8): patients presenting with a current relapse who had the history of ON in the previous relapses. Group III (n = 6): patients presenting with a current relapse but with no history of ON. Neuro ophtalmological evaluation. VEP investigation and orbital MRI with triple dose (0.3 mmol/kg) gadolinium (Gd) were carried out for all. VEP was found to be 70% sensitive and 12.5% specific to the acute ON, whereas orbital MRI with triple dose Gd was 70% sensitive and 100% specific. In chronic ON, the sensitivity of orbital MRI is 0%, whereas the VEP is still 75% sensitive to chronic optic nerve involvement and can distinguish the pathology 100% specifically. In conclusion, orbital MRI with triple dose Gd is not more sensitive than VEP in determining the acute optic nerve pathologies but it is a 100% specific method. The results suggest that VEP is superior to the orbital MRI in determining the chronic optic nerve involvement. PMID- 15527207 TI - Lithium ion "cyclotron resonance" magnetic fields decrease seizure onset times in lithium-pilocarpine seized rats. AB - The cyclotron resonance equation predicts that the frequency of an applied magnetic field that might optimally interact with a single ion species may be computed as a function of the charge-to-mass ratio of the ion and the strength of the background static magnetic field. The present study was undertaken to discern the applicability of this equation for optimizing lithium ion utilization in the rat, as inferred by the predicted magnetic "ion resonance "field-induced shift of lithium's dose-dependent curve for seizure onset times (SOTs) when combined with the cholinergic agent pilocarpine. Groups of rats were administered 1.5 thru 3 mEq/kg lithium chloride (in 0.5 mEq/kg increments) and exposed to reference conditions or to one of three intensities (70 nanoTesla, 0.8 microTesla, or 25 microTesla) of a 85 Hz magnetic field calculated to resonate with lithium ions given the background static geomagnetic field of approximately 38,000 nanoTesla (0.38 Gauss). A statistically significant quadratic relationship for SOT as a function of magnetic field intensity (irrespective of lithium dose) was noted: this U-shaped function was characterized by equal SOTs for the reference and 25 microTesla groups, with a trend toward shorter SOTs for the 70 nanoTesla and 0.8 microTesla groups. Although not predicted by the equations, this report extends other findings suggestive of discrete intensity windows for which magnetic field frequencies derived from the cyclotron ion resonance equation may affect ion activity. PMID- 15527208 TI - ["Second hand" experience. About hearing--saying]. PMID- 15527209 TI - [Dyspnea!--Managing difficult events in palliative nursing and hospice terminal care nursing]. PMID- 15527210 TI - [Occupational accidents can have psychiatric sequelae]. PMID- 15527211 TI - [Nursing visit as a means for quality assurance]. PMID- 15527212 TI - [Danger especially in home nursing by family]. PMID- 15527213 TI - [How is quality in home nursing assured?]. PMID- 15527214 TI - [ASB for the future of nursing in Germany]. PMID- 15527215 TI - ["Higher outcomes"--optimizing therapy of schizophrenia]. PMID- 15527216 TI - [Attention deficit hyperactivity syndrome (ADHS) in Germany]. PMID- 15527217 TI - [In children with psychiatrically ill parents the soul suffers]. PMID- 15527218 TI - [FDA approval for the antidepressive drug Cymbalta]. PMID- 15527219 TI - [Developments in therapy of malignant gliomas in the last 30 years]. PMID- 15527220 TI - [Changes in survival by combined radio-chemotherapy]. PMID- 15527221 TI - [Adjuvant therapy of colonic carcinoma--soon in tablet form]. PMID- 15527222 TI - [Tyrosine kinase inhibitor erlotinib (Tarceva) improves survival of patients with multiple previous treatments]. PMID- 15527224 TI - [Start of the new German Register for Neuroendocrine Tumors]. PMID- 15527223 TI - [Pancreatic carcinoma: progress in therapy]. PMID- 15527225 TI - [The most recent research results confirm the safety and effectiveness of iscador in cancer]. PMID- 15527226 TI - [The nursing uniform is often unsuitable]. PMID- 15527227 TI - [First antibody in oncology continues on a successful course]. PMID- 15527228 TI - [Diabetes mellitus: global strategies for control of the epidemic]. PMID- 15527229 TI - ["Living better--thanks to insulin"]. PMID- 15527231 TI - [Blood glucose self-monitoring is now simpler and more comfortable!]. PMID- 15527230 TI - [Novo Nordisk introduces a new basic insulin for marketing]. PMID- 15527232 TI - [Attitudes, wishes and notes of diabetic patients]. PMID- 15527233 TI - [Risk for patients often underestimated]. PMID- 15527234 TI - [Lantus is superior in BOT (basic oral therapy) to standard mixed insulin]. PMID- 15527235 TI - [Study proves anti-hepatitis A virus effectiveness of Korsolex endoscope disinfectant]. PMID- 15527236 TI - [Diagnostic imaging in children]. PMID- 15527237 TI - [Start of the "simply from love" initiative]. PMID- 15527238 TI - [Testosterone replacement in urology]. PMID- 15527239 TI - [Innovations improve quality of life of patients with hemophilia A]. PMID- 15527240 TI - [Psoriatic arthritis (PsA)]. PMID- 15527241 TI - [Multiple sclerosis--a challenge for physician and patient]. PMID- 15527242 TI - [Recognizing and treating malnutrition]. PMID- 15527243 TI - [Modern migraine therapy]. PMID- 15527244 TI - [Evidence-based therapy of intermittent claudication]. PMID- 15527245 TI - [Current EULAR data on valdecoxib. New study results of gastrointestinal safety and "cost effectiveness"]. PMID- 15527246 TI - [Separation into an unclean and a clean side--even when there is little space]. PMID- 15527247 TI - [Women and addictions, 3: Women and nicotine dependence]. PMID- 15527249 TI - ["Courage for loneliness": about dealing with yourself]. PMID- 15527248 TI - [Controlling stubborn prejudgment]. PMID- 15527250 TI - [Giving away for free at termination?]. PMID- 15527251 TI - [Deep sleep: enveloped by nurses]. PMID- 15527252 TI - Lihouidine, a novel spiro polycyclic aromatic alkaloid from the marine sponge Suberea n. sp. (Aplysinellidae, Verongida). AB - An investigation of a new species of sponge from the genus Suberea collected at Lihou Reef in the Coral Sea afforded lihouidine, an unprecedented cytotoxic spiro nonacyclic polyaromatic alkaloid. The structure of the alkaloid, which was racemic, was determined by a combination of 1D and 2D NMR techniques and single crystal X-ray structural analysis. PMID- 15527253 TI - One-pot synthesis of helical aromatics: stereoselectivity, stability against racemization, and assignment of absolute configuration assisted by experimental and theoretical circular dichroism. AB - Helical aromatics (1) were synthesized via one step in good quantity by solvent free condensation of N,N'-p-phenylenediamine (2) and various carboxylic acids in the presence of a Lewis acid. Microwave irradiation greatly facilitated the condensation reaction to furnish 1 with a 100% diastereo- and a 50% enantioselectivity, when a chiral carboxylic acid was utilized. 1f, derived from 2-methylglutaric acid, was quite stable, no racemization taking place even at 200 degrees C. The assignment of the absolute configurations to the helical aromatics has been achieved by experimental and theoretical CD spectra calculated by time dependent density functional theory. PMID- 15527254 TI - Formal synthesis of optically active ingenol via ring-closing olefin metathesis. AB - The construction of strained carbon skeletons by ring-closing olefin metathesis (RCM) was investigated. With well-designed diene 4, RCM was found to be applicable to the formation of a highly strained inside-outside bicyclo[4.4.1]undecane skeleton of ingenol, a bioactive diterpenoid, and formal total synthesis of optically active ingenol (1) was achieved. The key features of this synthesis are construction of an A-ring by spirocyclization of the ketone with an allylic chloride unit, 26, and ring closure of a B-ring by olefin metathesis. Starting from Funk's keto ester 6, the key intermediate aldehyde 9 in Winkler's total synthesis was synthesized in eight steps in 12.5% overall yield. This strategy of direct cyclization of a strained inside-outside skeleton provided the first easy access to optically active ingenol. PMID- 15527255 TI - Easy access to 3- or 5-heteroarylamino-1,2,4-triazines by S(N)Ar, S(N)H, and palladium-catalyzed N-heteroarylations. AB - In this paper, N-arylations between two heteroaryl compounds were studied. Conditions were found to generate selectively either 3- or 5-heteroarylamino 1,2,4-triazines by investigating anionic processes (use of bases such as 2,2',6,6'-tetramethylpiperidine/tBuOK/nBuLi) or Pd-catalyzed N-arylations [Pd(OAc)(2), xantphos]. These methods were successfully applied to a wide variety of heteroarylamines and allowed us to pursue our work on fused polynitrogen compounds synthesis. PMID- 15527256 TI - Combined directed ortho metalation/cross-coupling strategies: synthesis of the tetracyclic A/B/C/D ring core of the antitumor agent camptothecin. AB - A convergent synthesis of the A/B/C/D ring fragment 5 of camptothecin using a combination of directed ortho metalation and Negishi cross-coupling is described. The key features of the synthetic sequence are an anionic ortho-Fries rearrangement (10 --> 12), a Negishi cross-coupling (7 --> 6), and a terminal modified von Braun reaction (16 --> 5) that leads to tetracyclic derivative 5 in 7 steps and 11% overall yield. PMID- 15527257 TI - Stereoselective photochemical 1,3-dioxolane addition to 5-alkoxymethyl-2(5H) furanone: synthesis of bis-tetrahydrofuranyl ligand for HIV protease inhibitor UIC-94017 (TMC-114). AB - A convenient synthesis of (3R,3aS,6aR)-3-hydroxyhexahydrofuro[2,3-b]furan, a high affinity nonpeptidal ligand for HIV protease inhibitor UIC-94017, is described. This inhibitor is undergoing advanced clinical trials. The synthesis utilizes a novel stereoselective photochemical 1,3-dioxolane addition to 5(S) benzyloxymethyl-2(5H)-furanone as the key step. The requisite furanone derivative was prepared in high enantiomeric excess by an immobilized lipase-catalyzed selective acylation of (+/-)-1-(benzyloxy)-3-buten-2-ol and a ring-closing olefin metathesis with Grubbs' catalyst. Optically active bis-THF was converted to protease inhibitor 2 (UIC-94017). PMID- 15527258 TI - Microwave-assisted synthesis of highly substituted aminomethylated 2-pyridones. AB - By employing microwave-assisted organic synthesis (MAOS) efficient conditions to introduce aminomethylene substituents in highly substituted bicyclic 2-pyridones have been established. Primary amino methylene substituents were introduced via a cyanodehalogenation followed by a borane dimethyl sulfide reduction of the afforded nitrile. In both of these transformations, microwave irradiation proved to be superior to traditional conditions and the primary amines were obtained in good overall yields (55-58% over three steps). To incorporate tertiary aminomethylene substituents in the 2-pyridone framework, a microwave-assisted Mannich reaction using preformed iminium salts proved to be effective. Thus highly substituted 2-pyridones were obtained in 48-93% yields. PMID- 15527259 TI - Indole synthesis by controlled carbolithiation of o-aminostyrenes. AB - An effective synthesis of the functionalized indole ring system has been developed from substituted o-aminostyrene starting material. Our methodology involves a novel cascade reaction sequence of alkyllithium addition to the styrene double bond and subsequent trapping of the intermediate organolithium with a suitable electrophile, followed by an in situ ring closure and dehydration to generate the indole ring. This new reaction sequence allows for the introduction of molecular diversity at all positions on the indole scaffold. The procedure was shown to be successful with a range of both C and N substituents on the o-aminostyrenes. The reaction sequence was tolerant to the reactivity range of alkyllithiums such as tert-, sec-, and n-butyllithium. The electrophiles used were DMF, which generated indole products with C-2 unsubstituted, and nitriles, which incorporated the nitrile substituent at C-2. The o-aminostyrene starting materials were generated by a Pd-catalyzed cross-coupling reaction of a vinyl boronic acid equivalent with the readily available substituted o-bromoanilines. PMID- 15527260 TI - Highly stereoselective aldol reaction for the synthesis of gamma-lactones starting from tartaric acid. AB - A simple stereoselective process for the synthesis of highly substituted gamma lactones was developed based on aldol reactions between the enolate of dioxanes derived from tartaric acid and aldehydes. A range of aromatic and aliphatic aldehydes were reacted, in most cases achieving good yields and stereoselectivity. The limitations of this reaction were identified and a transition state is proposed. PMID- 15527261 TI - A practical synthesis of gramicidin s and sugar amino Acid containing analogues. AB - A practical gram-scale and high-yielding synthesis of the antimicrobial peptide gramicidin S is presented. An Fmoc-based solid-phase peptide synthesis protocol is employed for the generation of the linear decapeptide precursor, which is cyclized in solution to afford the target compound. The versatility of our method is demonstrated by the construction of eight gramicidin S analogues (15a-h) having nonproteinogenic sugar amino acid residues (4-7) incorporated in the turn regions. PMID- 15527262 TI - Triplet-sensitized photolysis of the photoisomer of a 2,11 diaza[3,3](9,10)anthracenoparacyclophane: an adiabatic cycloreversion and a [2pia + 2pia + 2sigmas] rearrangement in a triplet state of the biplanophane system. AB - The triplet-sensitized photoreactions of the title biplanophane system 6, the photoisomer of a 2,11-diaza[3,3](9,10)anthracenoparacyclophane derivative 5, were investigated by stationary and laser-flash photolyses using xanthone (XT) and benzophenone (BP) as triplet sensitizers. When photoisomer 6 underwent XT sensitized irradiation, a triplet cyclophane 5 and a novel polycyclic product 7 were obtained via an adiabatic cycloreversion and a formal [2pia + 2pia + 2sigmas] rearrangement, respectively. The maximum quantum yield for the formation of cyclophane 5 (0.69) and the upper-limit efficiency for the formation of polycycle 7 (0.31) were determined by laser photolysis techniques. For BP sensitized photolysis of photoisomer 6, oxetane 8, in addition to triplet cyclophane 5 and polycycle 7, was formed by a Paterno-Buchi reaction. The quenching rate constant (k(q)) of triplet BP by photoisomer 6 (3.4 x 10(8) dm(3) mol(-)(1) s(-)(1)) was found to be 1 order of magnitude smaller than that for XT (5.0 x 10(9) dm(3) mol(-)(1) s(-)(1)). On the basis of the relationship between k(q) and the triplet donor-acceptor energy gap, the triplet energy level of photoisomer 6 was estimated to be approximately 71 kcal mol(-)(1). The photochemical and the photophysical processes involved in the sensitized photolyses are summarized in an energetic reaction diagram and discussed in detail. PMID- 15527263 TI - Modulation of chemoselectivity by protein additives. Remarkable effects in the oxidation of hyperforin. AB - Protein additives have a dramatic effect on the H(2)O(2) oxidation of hyperforin, either protecting the enolized phloroglucinol core from oxidation (human albumin) or promoting (HRP and ovalbumin) reaction pathways derived from the intermediacy of the enollactone 4, a minor component of the oxidation mixture in the absence of protein additives. To rationalize the exquisite specificity of several steps and their mechanistic oddity, an organocatalytic effect is postulated. The use of protein additives allows a straightforward and multigram preparation of the enollactone 6, an interesting multifunctionalized scaffold for bioactivity induction and/or modulation. PMID- 15527264 TI - Full stereochemical assignment and synthesis of the potent anthelmintic pyrrolobenzoxazine natural product CJ-12662. AB - The structure of the unusual anthelmintic pyrrolobenzoxazine terpenoid natural product CJ-12662 was established by X-ray crystallography and partial synthesis from 2-chloronitrobenzene. An unusual Meisenheimer-type rearrangement was used to provide the core pyrrolobenzoxazine heterocycle, and coupling of a tetracyclic pyrrolobenzoxazine lactone with the terpene alcohol was used to complete the synthesis of CJ-12662. PMID- 15527265 TI - Microwave-assisted acylation of amines, alcohols, and phenols by the use of solid supported reagents (SSRs). AB - A microwave-assisted synthesis of solid-supported reagents for the acylation of amines has been developed, and the same methodology has been successfully applied to the preparation of acylating agents anchored on different solid supports. Similarly, alcohols, phenols, and thiophenols have been easily acylated using these reagents under microwave irradiation. PMID- 15527266 TI - Tropiporphyrins, cycloheptatrienyl analogues of the porphyrins: synthesis, spectroscopy, chemistry, and structural characterization of a silver(III) derivative. AB - Tripyrranes were condensed with 1,3,5-cycloheptriene-1,6-dicarbaldehyde in TFA CH(2)Cl(2) to give, following oxidation with 0.1% aqueous ferric chloride solutions, a series of tropiporphyrins 9. These cycloheptatrienyl analogues of the porphyrins show strong diatropic ring currents by proton NMR spectroscopy where the internal CH gives a resonance at -7.3 ppm, although the meso-protons are not shifted as far downfield as most aromatic porphyrinoid systems. These data indicate that the seven-membered ring distorts the porphyrinoid macrocycle and decreases the overall diatropicity in tropiporphyrins. Addition of trace amounts of TFA to solutions of 9 affords the corresponding aromatic monocations, and at higher acid concentrations a nonaromatic dication is generated. The dication has undergone C-protonation at one of the meso-bridges and has lost the plane of symmetry present in the parent system. This species shows significant downfield shifts to the cycloheptatrienyl protons, indicating that this unit has taken on tropylium character. Tropiporphyrin 9a underwent a Diels-Alder cycloaddition with dimethyl acetylenedicarboxylate in refluxing xylenes to give modest yields of the related adduct. The Diels-Alder adduct 17 showed an increased diatropic ring current where the internal proton shifted beyond -9 ppm, and this indicates that the [18]annulene substructure has flattened out compared to 9a. Diimide reduction of 9a afforded a dihydrotropiporphyrin that also showed a stronger ring current. Tropiporphyrins 9 were also shown to react with silver(I) acetate in the presence of DBU in refluxing pyridine to give the corresponding silver(III) organometallic derivatives. The meso-protons for these metal complexes give proton NMR chemical shift values similar to those for the parent tropiporphyrins, indicating that the macrocycle is still distorted, but the external olefinic protons are shifted downfield compared to 9. A diphenyl substituted silver(III) derivative 18b was further characterized by X-ray crystallography. This shows that the cycloheptatriene unit takes on a highly twisted geometry that distorts the overall conformation of the porphyrinoid macrocycle. PMID- 15527267 TI - Reaction of bis(2,4-dinitrophenyl) phosphate with hydrazine and hydrogen peroxide. Comparison of O- and N- phosphorylation. AB - Nonionic hydrazine reacts with anionic bis(2,4-dinitrophenyl) phosphate (BDNPP), giving 2,4-dinitrophenyl hydrazine and dianionic 2,4-dinitrophenyl phosphate by an S(N)2(Ar) reaction, and at the phosphoryl center, giving 2,4-dinitrophenoxide ion and a transient phosphorylated hydrazine that rearranges intramolecularly to N-(2,4-dinitrophenyl)-N-phosphonohydrazine. Approximately 58% of the reaction at pD = 10 occurs by N-phosphorylation, as shown by (31)P NMR spectroscopy. Reaction of HO(2)(-) is wholly at phosphorus, and the intermediate peroxophosphate reacts intramolecularly, displacing a second 2,4-dinitrophenoxide ion, or with H(2)O(2), giving 2,4-dinitrophenyl phosphate and O(2). Rate constants of O- and N phosphorylation in reactions at phosphorus of NH(2)NH(2), HO(2)(-), and NH(2)OH and its methyl derivatives follow Bronsted relationships with similar slopes, but plots differ for oxygen and nitrogen nucleophiles. The reaction with NH(2)NH(2) has been probed by using both NMR spectroscopy and electrospray ionization mass and tandem mass spectrometry, with the novel interception of key reaction intermediates in the course of reaction. PMID- 15527268 TI - Asymmetric iodolactamization induced by chiral oxazolidine auxiliary. AB - Asymmetric iodolactamization reactions of unsaturated amides with oxazolidines as the chiral auxiliaries were investigated. With (4S)-4-((2R)-2-butyl)-2,2 dimethyloxazolidine as the auxiliary and LiH as the base, a number of unsaturated amides underwent iodolactamization smoothly to afford the corresponding gamma- and delta-lactams in 30-98% yield with de values up to 97%. PMID- 15527269 TI - Highly enantioselective cyanosilylation of aldehydes catalyzed by novel beta amino alcohol-titanium complexes. AB - The beta-amino alcohol 1b-Ti(Oi-Pr)(4) complex has been shown to catalyze the enantioselective cyanosilylation of aldehydes efficiently. In the presence of 5 mol % of 1b-Ti(Oi-Pr)(4) complex catalyst, the aromatic, conjugated, heteroaromatic, and aliphatic aldehydes were converted to their corresponding trimethylsilyl ethers of cyanohydrins in 90-99% yields with up to 94% ee under mild conditions. PMID- 15527270 TI - Selective lithiation of 4-(1H-1-pyrrolyl)pyridine. access to new electron releasing ligands. AB - The first lithiation of 4-(1H-1-pyrrolyl)pyridine has been realized. The use of BuLi-containing lithium aggregates induced the selective pyridine ring functionalization by taking advantage of the electron-donor effect of the pyrrole nucleus. Opportune substituents were introduced alpha to the pyridine nitrogen leading to new electron-enriched pyridylphosphine, bipyridine, and terpyridine ligands. PMID- 15527271 TI - Aqueous-phase, palladium-catalyzed cross-coupling of aryl bromides under mild conditions, using water-soluble, sterically demanding alkylphosphines. AB - Sterically demanding, water-soluble alkylphosphines have been used in combination with various palladium salts in Suzuki, Sonogashira, and Heck couplings of aryl bromides under mild conditions in aqueous solvents. The tert-butyl-substituted ligands 2-(di-tert-butylphosphino)ethyltrimethylammonium chloride (t-Bu-Amphos) and 4-(di-tert-butylphosphino)-N,N-dimethylpiperidinium chloride (t-Bu-Pip-phos) in combination with palladium(II) salts were found to give catalysts that were significantly more active than catalysts derived from tri(3 sulfonatophenyl)phosphine trisodium (TPPTS). Suzuki couplings of unactivated aryl bromides occurred efficiently at room temperature in water/acetonitrile and water/toluene biphasic mixtures or in neat water. Notably, Suzuki couplings of hydrophilic aryl bromides gave high yields without using organic solvents for the reaction or purification. This methodology has been applied to a highly efficient synthesis of diflunisal. The catalyst derived from t-Bu-Amphos was recycled three times in Suzuki couplings in water/toluene before catalyst activity began to significantly drop. The average yield of four cycles was >80% per cycle. Heck and Sonogashira couplings were carried out under mild conditions (50 and 80 degrees C, respectively) with unactivated aryl bromides to give coupled products in high yield. PMID- 15527272 TI - Alkynyliodonium salts in organic synthesis. Application to the preparation of the tricyclic core of (+/-)-halichlorine. AB - The tricyclic core of halichlorine has been synthesized through the use of an alkynyliodonium salt/alkylidenecarbene/1,5 C-H insertion sequence that sets the pivotal quaternary center in the target. PMID- 15527273 TI - Preparation and characterization of regioisomerically pure 1,7-disubstituted perylene bisimide dyes. AB - A detailed study on bromination and subsequent imidization of perylene bisanhydride with cyclohexylamine is reported. The present results reveal that previously reported 1,7-difunctionalized perylene bisimides are presumably contaminated with the respective 1,6 regioisomers. N,N'-Dicyclohexyl-1,7 dibromoperylene bisimide 1,7-3 is obtained for the first time in isomerically pure form, and its structure is unequivocally confirmed by X-ray analysis. By using regioisomerically pure 1,7-dibromoperylene bisimide 1,7-3, 1,7 dipyrrolidinylperylene bisimides 4a-c and 1,7-dipyrrolidinylperylene bisanhydride 5 as well as the unsymmetrically difunctionalized 1-bromo-7-pyrrolidinyl- and 1 cyano-7-pyrrolidinylperylene bisimides 7 and 8 are synthesized in good yield. PMID- 15527274 TI - Asymmetric synthesis of 3-substituted proline chimeras bearing polar side chains of proteinogenic amino acids. AB - The amino-zinc-ene-enolate cyclization reaction is a straightforward route to the synthesis of 3-substituted prolines. Herein we report the application of this reaction to the syntheses of proline chimeras of lysine, glutamic acid, glutamine, arginine, and serine. All these compounds were obtained in enantiomerically pure form and suitably protected for peptide synthesis. PMID- 15527275 TI - Stereocontrolled total synthesis of a polyfunctional carotenoid, peridinin. AB - Peridinin, which was isolated from the planktonic algae dinoflagellates causing red tides, is a highly oxidized carotenoid containing an allene and a characteristic (Z)-gamma-ylidenebutenolide function in the main conjugated polyene chain in addition to functionalized cyclohexane rings at both ends of the molecule. We achieved a stereocontrolled total synthesis of peridinin by featuring the Sharpless asymmetric epoxidation under precise reaction conditions, Wittig reaction with silylfuranmethylide followed by photosensitized oxygenation, stereocontrolled Pd-catalyzed one-pot (Z)-gamma-ylidenebutenolide synthesis, and modified Julia-Kocienski olefination. This synthesis is the first example of controlling the stereochemistry of polyfunctional allenic carotenoids. PMID- 15527276 TI - Enantioselective allylic substitution of cinnamyl esters catalyzed by iridium chiral aryl phosphite complex: conspicuous change in the mechanistic spectrum by a countercation and solvent. AB - Iridium-catalyzed asymmetric allylic alkylation of monoaryl substrates 4-6 with chiral phosphites 1-3 has been investigated. Although branched isomers were formed with high regioselectivities, the enantioselectivities of these products were remarkably influenced by solvents, countercations, and additives (ZnCl(2) and LiCl). PMID- 15527277 TI - Toward the development of a cephalosporin-based dual-release prodrug for use in ADEPT. AB - In previous work we have shown that a cephalosporin structure bearing an S aminosulfenimine at the 7-position behaved as a beta-lactamase-dependent dual release prodrug. Scission of the beta-lactam ring of such a structure led to the rapid loss of the sulfur-attached side chain moiety via an intramolecular displacement, while the 3'-group was lost via the well-established elimination process at that position. In the present work we report on an evaluation of the scope and limitations of exploiting the S-aminosulfenimine functionality to generate a cephalosporin-based prodrug incorporating two biologically active components. Starting from 7-ACA, a viable synthetic cycle was put in place that avoided formation of the Delta(2) isomer throughout and that allowed incorporation of aminoglutethimide at the 3'-position and of a tosyl S aminosulfenimine at the 7-position. The direct incorporation of a biologically active sulfonamide (ethoxzolamide) or a sulfamate (coumate) at this latter position was not achieved as a result of the difficulty of generating the corresponding sulfur diimides. An indirect route for the formation of an S aminosulfenimine was put in place, as was a general method of alkylation (Mitsunobu reaction) of the tosyl S-aminosulfenimine following its incorporation. PMID- 15527278 TI - Stereoselective Rh-catalyzed hydrogenation of cyclobutyl chiral enamides: double stereodifferentiation vs catalyst-controlled diastereoselection. AB - The hydrogenation reactions of several cyclobutyl enamides derived from (-)-alpha pinene or (-)-verbenone have been investigated by using different catalysts. The chiralities of both the substrate and the catalyst as well as the Z/E stereochemistry of the double bond have been considered, and the observed diastereoselectivity has been rationalized. For enamides with the double bond separated from the cyclobutane by a methylene, the Wilkinson catalyst did not induce any diastereoselection, but excellent diastereoselectivity was observed when using Et-DuPHOS-Rh and ChiraPHOS-Rh. The configuration of the new stereogenic center was catalyst-dependent and can be rationalized according to the Halpern mechanism. For (Z)-enamides with the double bond directly linked to the cyclobutane ring, the chirality of the substrate governed the diastereoselection and the Halpern mechanism seemed not to be operative in the hydrogenation with ChiraPHOS, with the configuration of the new stereogenic center being determined by steric effects. On the contrary, the chirality of the catalyst was the factor determining the stereochemistry of the major products with alkyl-DuPHOS-Rh. Z/E stereochemistry influenced the stereodifferentiation, and a different behavior for each Z or E stereoisomer was found. For both (Z)- and (E)-enamides, some instances of match/mismatch between the chirality of the substrate and that of the catalyst were observed. As a result of all of these studies, a series of new cyclobutyl alpha-amino acids has been synthesized. These products are interesting to incorporate into conformationally constrained peptides. PMID- 15527279 TI - Anion radical [2 + 2] cycloaddition as a mechanistic probe: stoichiometry- and concentration-dependent partitioning of electron-transfer and alkylation pathways in the reaction of the Gilman reagent Me2CuLi.LiI with bis(enones). AB - Exposure of easily reduced aromatic bis(enones) 1a-1e to the methyl Gilman reagent Me(2)CuLi.LiI at 0 degrees C in tetrahydrofuran solvent provides the products of tandem conjugate addition-Michael cyclization, 2a-2e, along with the products of [2 + 2] cycloaddition, 3a-3e. Complete partitioning of the Gilman alkylation and [2 + 2] cycloaddition pathways may be achieved by adjusting the loading of the Gilman reagent, the rate of addition of the Gilman reagent, and the concentration of the reaction mixture. The Gilman alkylation manifold is favored by the rapid addition of excess Gilman reagent at higher substrate concentrations, while the [2 + 2] cycloaddition manifold is favored by slow addition of the same Gilman reagent at lower concentrations and loadings. Notably, [2 + 2] cycloaddition to form 3a-3e is catalytic in Gilman reagent. Kinetic data reveal that the ratio of 2a and 3a changes such that the cycloaddition pathway becomes dominant upon increased consumption of Gilman reagent. These data suggest a concentration-dependent speciation of the Gilman reagent and differential reactivity of the aggregates present at higher and lower concentrations. While the species present at higher concentration induce Gilman alkylation en route to products 2a-2e, the species present at lower concentration provide products of catalytic [2 + 2] cycloaddition, 3a-3e. Moreover, upon electrochemical reduction of the bis(enones) 1a-1e, or chemically induced single electron transfer from arene anion radicals, the very same [2 + 2] cycloadducts 3a-3e are formed. The collective data suggest that [2 + 2] cycloadducts 3a-3e arising under Gilman conditions may be products of anion radical chain cyclobutanation that derive via electron transfer (ET) from the Me(2)CuLi.LiI aggregate(s) present at low concentration. These observations provide a link between the Gilman alkylation reaction and related ET chemistry and suggest these reaction paths are mechanistically distinct. This analysis is made possible by the recent observation that easily reduced bis(enones) are subject to intramolecular [2 + 2] cycloaddition upon cathodic reduction or chemically induced ET from arene anion radicals, and is herewith showcased as a novel method of testing for the intermediacy of enone anion radicals. PMID- 15527280 TI - A facile and efficient synthesis of (Purin-6-yl)alanines. AB - (Purin-6-yl)alanines, a new class of amino acid-nucleobase conjugates, were synthesized by palladium-catalyzed cross-coupling reactions of protected iodozincalanines with 6-iodopurines (9-Bn-6-iodopurine and 9-THP-6-iodopurine as well as acyl-protected 6-iodopurine ribonucleoside and 2-deoxyribonucleoside). Free purine base and nucleosides bearing alanine in position 6 were obtained after complete deprotection of the products of cross-coupling reactions. PMID- 15527281 TI - Synthesis of a new class of furan-fused tetracyclic compounds using o quinodimethane chemistry and investigation of their antiviral Activity. AB - The synthesis and evaluation of antiviral activity of new furan-fused tetracyclic compounds are described. The syntheses were satisfactorily achieved on the basis of o-quinodimethane chemistry, using furan-containing benzocyclobutene derivatives as a substrate, in high generality and stereoselectivity. The various derivatives thus synthesized were examined on their inhibitory activity on virus growth using a hemagglutinin (HA) method, leading to a discovery of promising candidates for new antiviral drugs having high activity and good therapeutic index. PMID- 15527282 TI - Spin trapping by 5-carbamoyl-5-methyl-1-pyrroline N-oxide (AMPO): theoretical and experimental studies. AB - The nitrone 5-carbamoyl-5-methyl-1-pyrroline N-oxide (AMPO) was synthesized and characterized. Spin trapping of various radicals by AMPO was demonstrated for the first time by electron paramagnetic resonance (EPR) spectroscopy. The resulting spin adducts for each of these radicals gave unique spectral profiles. The hyperfine splitting constants for the superoxide adduct are as follows: isomer I (80%), a(nitronyl)(-)(N) = 13.0 G and a(beta)(-)(H) = 10.8 G; isomer II (20%), a(nitronyl)(-)(N) = 13.1 G, a(beta)(-)(H) = 12.5 G, and a(gamma)(-)(H) = 1.75 G. The half-life of the AMPO-O(2)H was about 8 min, similar to that observed for EMPO but significantly shorter than that of the DEPMPO-O(2)H with t(1/2) approximately 16 min. However, the spectral profile of AMPO-O(2)H at high S/N ratio is distinguishable from the spectrum of the (*)OH adduct. Theoretical analyses using density functional theory calculations at the B3LYP/6 31+G//B3LYP/6-31G level were performed on AMPO and its corresponding superoxide adduct. Calculations predicted the presence of intramolecular H-bonding in both AMPO and its superoxide adduct. The H-bonding interaction was further confirmed by an X-ray structure of AMPO, and of the novel and analogous amido nitrone 2 amino-5-carbamoyl-5-methyl-1-pyrroline N-oxide (NH(2)-AMPO). The thermodynamic quantities for superoxide radical trapping by various nitrones have been found to predict favorable formation of certain isomers. The measured partition coefficient in an n-octanol/buffer system of AMPO was similar to those of DMPO and DEPMPO. This study demonstrates the suitability of the AMPO nitrone for use as a spin trap to study radical production in aqueous systems. PMID- 15527283 TI - Activation of carbon dioxide by bicyclic amidines. AB - Activation of the carbon dioxide molecule was achieved using bicyclic amidines (DBU, PMDBD, and DBN). The solution reaction of CO(2) with amidines yielded the corresponding zwitterionic complexes through the formation of a N-CO(2) bond. (13)C NMR data confirmed the carbamic nature of the carbamic zwitterions, DBU CO(2) and PMDBD-CO(2). However, when these adducts were crystallized, the X-ray analyses of the single crystals were in agreement with bisamidinium bicarbonate salt structures, indicating that structural changes occurred in the crystallization process. The elemental and thermogravimetric analysis data for the carbamic zwitterions, DBU-CO(2) and PMDBD-CO(2), initially obtained by the direct reaction of amidines with CO(2), suggest that these molecules are probably associated with one molecule of water by hydrogen-bond formation (amidinium(+) COO(-)...H(2)O). A correlation was observed between the thermal stability and the transcarboxylating activity for the amidine-CO(2) complexes. Theoretical calculations of hardness were performed at the B3LYP/cc-pVTZ level of theory and showed concordance with the experimental reactivity of DBU and PMDBD toward CO(2). PMID- 15527284 TI - Synthesis and hydrolysis of a cis-chlorohydrin derived from a benzo[a]pyrene 7,8 diol 9,10-epoxide. AB - (+/-)-7beta,8alpha-Dihydroxy-9beta,10beta-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (DE-1) undergoes reaction with anhydrous HCl in dioxane to yield predominantly ( approximately 94%) a single chlorohydrin. This chlorohydrin was assigned structure 9, in which the chloro goup at C-10 is located cis to the C-9 hydroxyl group, on the basis of its (1)H NMR spectrum. This result is in contrast to the reaction of a diastereomeric benzo[a]pyrene 7,8-diol 9,10-epoxide (DE-2) with HCl, which yields only trans-chlorohydrin 8. The hydrolysis of cis-chlorohydrin 9 in 10:90 dioxane-water solutions yields the same ratio of tetrols ( approximately 89% cis/11% trans) as that formed by acid-catalyzed hydrolysis of DE-1. This result again contrasts with the hydrolysis of trans-chlorohydrin 8, which undergoes hydrolysis to give tetrols in a ratio different from that from acid catalyzed hydrolysis of DE-2. A marked common ion rate depression in the hydrolysis of cis-chlorohydrin 9 is observed, which shows that hydrolysis proceeds via an intermediate carbocation that has a sufficient lifetime to be trapped by external chloride ion. The observation that DE-1 reacts with HCl to give mainly the cis-chlorohydrin is rationalized by quantum chemical calculations that suggest that the cis-chlorohydrin is more stable than the epimeric trans chlorohydrin. PMID- 15527285 TI - A theoretical investigation on the mechanism of the PtCl2-mediated cycloisomerization of heteroatom-tethered 1,6-enynes. AB - A mechanistic study based on DFT theoretical calculations for the PtCl(2) catalyzed formation of bicyclic adducts from heteroatom tethered 1,6-enynes is reported. Different reaction pathways have been taken into account and the results are discussed. This analysis clearly reveals that the kinetically preferred pathway involves an initial 6-endo-cyclization from a triggered reactant complex by pi-complexation of Pt(II) onto the alkyne to form a cyclopropyl platina-carbene intermediate, followed by a [1,2]-hydrogen shift. PMID- 15527286 TI - Computational studies on the electrocyclizations of 1-amino-1,3,5-hexatrienes. AB - Electrocyclizations of 1,3,5-hexatrienes containing up to four electron-donating and/or electron withdrawing substituents have been studied computationally using the hybrid density functional, B3LYP. Electron donating substituents at positions C-1 and C-5 decrease activation barriers by 0.3 to 2.3 kcal/mol. Introducing of an electron-withdrawing group, CO(2)Me, at C-4 further decreases the activation energy by 7 kcal/mol. Electron-withdrawing groups (NO(2), SO(2)Ph and C=N(+)Me(2)) at C-2 have a profound effect of 17-25 kcal/mol on the activation energy. PMID- 15527287 TI - Toward nanoamphiphiles: efficient synthesis of desymmetrized polyphenylene dendrimers. AB - A new synthetic approach for the desymmetrization of polyphenylene dendrimers (PPDs) is described. Tetrakis(4-ethynylphenyl)methane undergoes facile Diels Alder cycloaddition with substoichiometric quantities of tetraphenylcyclopentadienones bearing one polar functional group. A single ethynyl group is thereby converted to a rigid, selectively functionalized polyphenylene moiety, which serves as a focal point for further transformations or interfacial anchoring. This is the key feature for the design of desymmetrized monodisperse macromolecules with a spherical shape. The remaining unreacted ethynyl groups provide a trifold core for the stepwise elaboration of first- and second-generation polyphenylene dendrons, which may, in turn, bear specific numbers of different peripheral functional groups at their terminae. Moreover, the resulting macromolecules exhibit the characteristic shape-persistence and monodispersity of PPDs. This approach is an important achievement in nanosciences, especially for tailoring new nanoamphiphiles. It is also of synthetic importance, as it enables the separation of two regioisomeric polyphenylene dendrimers for the first time. PMID- 15527288 TI - A new series of pyrimidine-containing linear molecules: their elegant crystal structures and intriguing photophysical properties. AB - A new series of aza-substituted analogues 3-5 based on the 1,4 bis(phenylethynyl)benzene moiety have been synthesized by the selective Pd catalyzed Sonogashira coupling reaction from 5-bromo-2-iodopyrimidine (1). In these linear molecules, the dipolar pyrimidine moiety is introduced as a probe to investigate factors that control the intermolecular interactions over the crystal engineering. The results reveal that the manner of packing changes both dipolar interactions between linear pyrimidine-containing molecules and transition moments simultaneously, resulting in remarkably different photophysical properties. Due to their versatile dipole-dipole and face-to-face pi piinteractions in a crystal motif, further applications on the design of ordered crystalline materials for the field effect transistors are promising. PMID- 15527289 TI - Highly enantioselective conjugate additions of potassium organotrifluoroborates to enones by use of monodentate phosphoramidite ligands. AB - The use of phosphoramidite ligands in the rhodium-catalyzed asymmetric conjugate addition of potassium organotrifluoroborates to various enones in the absence of water is described. A systematic search for effective catalysts has been performed by use of high-throughput screening methods. Initially, we have screened reaction conditions, catalyst precursors, and focused ligand libraries. In the next stage we have used the monodentate ligand combination approach, and finally we have made a library of 96 different phosphoramidites by parallel synthesis in the robot (instant ligand libraries) and have tested these in the vinylation of cyclohexenone (up to 88% enantiomeric excess, ee) and 4-phenyl-3 buten-2-one (up to 42% ee). Arylation of cyclohexenone by use of potassium phenyltrifluoroborate gave 3-phenylcyclohexanone with 99% ee. PMID- 15527290 TI - PuPHOS: a synthetically useful chiral bidentate ligand for the intermolecular Pauson-Khand reaction. AB - Here we describe the synthesis and use of the Pulegone-derived bidentate P,S ligands PuPHOS and CyPuPHOS in the intermolecular Pauson-Khand reaction. Ligand exchange reaction of hexacarbonyldicobalt-alkyne complexes with PuPHOS provides a diasteromeric mixture of complexes (up to 4.5:1) from which the major isomers can be conveniently separated by simple crystallization. An isomerization crystallization sequence of the original mixture results in a dynamic resolution that allows the preparation of the pure major Co(2)(mu-TMSC(2)H)(CO)(4)-PuPHOS (15a) in a multigram scale. Pauson-Khand reaction of 15a with norbornadiene provided, for the first time, the corresponding enone 18 with up to 93% yield and 97% ee. The use of (+)-18 as a surrogate of chiral cyclopentadienone is also demonstrated. Copper-catalyzed Michael addition of a Grignard reagent followed by removal of the TMS group with TBAF were the most reliable methods to transform (+)-18 into valuable starting materials 20a-e for the enantioselective synthesis of cyclopentenoid systems. PMID- 15527291 TI - Synthesis and application of chiral cyclopropane-based ligands in palladium catalyzed allylic alkylation. AB - A series of chiral, cyclopropane-based phosphorus/sulfur ligands have been synthesized and evaluated in the palladium-catalyzed allylic alkylation of 1,3 diphenylpropenyl acetate with dimethyl malonate. Variation of the ligand substituents at phosphorus, sulfur, and the carbon backbone revealed 24d to have the optimal configuration for this reaction, giving the product in high yield and with good enantioselectivity (93%). A model for the observed enantioselectivity is discussed within the context of existing models, using X-ray crystallographic data, solution-phase NMR studies, and the absolute stereochemistry of the products. Selected ligands were also evaluated in the palladium-catalyzed intermolecular Heck reaction and the rhodium-catalyzed hydrogenation of a dehydroamino acid. PMID- 15527292 TI - Palladium-catalyzed reaction of boronic acids with chiral and racemic alpha-bromo sulfoxides. AB - Palladium-catalyzed cross-coupling reactions of racemic alpha-bromo sulfoxides with boronic acids are carried out in either aqueous or nonaqueous medium with formation of a new C sp(3)-C sp(2) bond. The arylation of chiral alpha-bromo sulfoxides occurs without racemization. The cross-coupling reaction is general and gives high yields with arylboronic acids substituted with either donor or acceptor groups but gives poor results with heteroarylboronic acids. The best yields are obtained using degassed solvents and CsF instead of aqueous base. The use of aqueous base and the presence of oxygen favor the homocoupling side reaction. PMID- 15527293 TI - Catalysis with phosphine-containing amino acids in various "turn" motifs. AB - We have been actively involved in the development of parallel approaches for the discovery of phosphine ligands. Our approach has been based on the incorporation of phosphine-containing amino acids into peptide sequences that are designed to have stable secondary structures. We have examined helical and turn secondary structures and have reported that alkylation of cyclopentenyl acetate with dimethylmalonate can be catalyzed in high enantiomeric excess (ee) with a beta turn-based ligand. The importance of the peptide secondary structure was demonstrated through the synthesis of a series of peptide ligands where the nature of the turn-forming residues was probed. Additionally, other turn-forming units and a variety of different phosphine-containing amino acids have been examined for their ability to control the selectivity of the allylation reaction. This paper reports the results obtained through the examination of different turn motifs as well as different phosphine substitutions on the "best" turn sequence, Pps-Pro-d-Xxx-Pps. PMID- 15527294 TI - Synthesis and electrogenerated chemiluminescence of donor-substituted phenylethynylcoumarins. AB - Two series of donor-bearing phenylethynylcoumarins have been synthesized, and their photophysical properties have been evaluated. Chemiluminescence was observed through the annihilation of their electrogenerated radical ions and was found to be only slightly affected by the presence of various donor groups on the phenyl moiety linked through the C-C triple bond. The overall properties of the two series of compounds are discussed with respect to their structures. The observed electronic absorption properties are explained with the help of computational studies. PMID- 15527295 TI - Dicyclobuta[de,ij]naphthalene and dicyclopenta[cd,gh]pentalene: a theoretical study. AB - The structures, energetics, and aromatic character of dicyclobuta[de,ij]naphthalene, 1, dicyclopenta[cd,gh]pentalene, 2, dihydrodicyclobuta[de,ij]naphthalene, 3, and dihydrocyclopenta[cd,gh]pentalene, 4, have been examined at the B3LYP/6-311++G//B3LYP/6-31G level of theory. All molecules are bowl-shaped, and the pentalene isomers, 2 and 4, are most stable. A comparison with other C(12)H(6) and C(12)H(8) isomers indicates that 2 is approximately 25 kcal/mol less stable than 1,5,9-tridehydro[12]annulene and 4 is approximately 100 kcal/mol higher in energy than acenaphthylene, both of which are synthetically accessible. The transition state structure for bowl-to-bowl inversion of 1 is planar (D(2)(h)()) and lies 30.9 kcal/mol higher in energy than the ground state; the transition state for inversion of 2 is C(2)(h)() and lies 46.6 kcal/mol higher in energy. Symmetry considerations, bond length alternations, and NICS values (a magnetic criterion) all indicate that the ground states of 1, 3, and 4 are very aromatic; however, HOMA values (a measure of bond delocalization) indicate that 3S and 4S are aromatic but that 1S is less so. NICS values for the ground state of 2 strongly indicate aromaticity; however, bond localization, symmetry, and HOMA values argue otherwise. PMID- 15527296 TI - Monomeric cobalt oxazoline palladacycles (COP). Useful catalysts for catalytic asymmetric rearrangement of allylic trichloroacetimidates. AB - Cobalt oxazoline palladacycles (COP) containing acetylacetonate and hexafluoroacetylacetonate ligands were prepared as catalysts for the asymmetric rearrangement of allylic trichloroacetimidates. These monomeric catalysts are more soluble than the previously described chloride-bridged dimer COP-Cl (1). COP hfacac (2) provides rearranged allylic trichloroacetamides with high enantiomeric purities (91-98% ee) in solvents of widely varying polarities: cyclohexane, toluene, 1,2-dichloroethane, ethyl acetate, acetone, acetonitrile, and THF. The first single-crystal X-ray structure of a COP catalyst is also reported. PMID- 15527297 TI - Mizoroki-Heck arylation of alpha,beta-unsaturated acids with a hybrid fluorous ether, F-626: facile filtrative separation of products and efficient recycling of a reaction medium containing a catalyst. AB - The Mizoroki-Heck reaction was carried out using a fluorous ether F-626 as the solvent and a fluorous Pd carbene complex as the catalyst. When carboxylic acids are the products, separation of both the F-626 and the Pd catalyst from the products can be conveniently carried out by simple filtration. The F-626 filtrates containing the Pd catalyst can be recycled. PMID- 15527298 TI - Reactive dications: the superacid-catalyzed reactions of alkynes bearing adjacent N-heterocycles or amine groups. AB - A variety of aminoalkynes and related heterocycles are reacted in the Bronsted superacid CF(3)SO(3)H (triflic acid), and products are obtained in generally good yields (69-99%) from Friedel-Crafts-type reactions. The reactions are consistent with the formation of novel dicationic intermediates having a vinyl cationic site and an adjacent protonated N-heterocycle or ammonium cation. PMID- 15527299 TI - Density functional theory study on the effect of substitution and ring annelation to the rim of corannulene. AB - B3LYP/6-311G calculations indicate that annelation of a five-membered ring to the rim of corannulene and substitution to all the rim carbons lowers the barrier for bowl-to-bowl inversion. Singlet-triplet energy differences, frontier orbital analysis, and nucleus-independent chemical shift (NICS) values indicate significant enhancement of the reactivity when the substitutions involve exocyclic double bonds. Bowl-to-bowl inversion barrier, curvature, and reactivity for unsaturated and saturated five-membered ring-annelated corannulenes are analogous to decamethyl- and decamethylene-substituted corannulenes. PMID- 15527300 TI - A scalable synthesis of a histamine H3 receptor antagonist. AB - Starting from 1-methylimidazole, a concise, scalable, three-step synthesis of the title compound is described. The required 2-chloroimidazole was prepared in very good yield by halogen-metal exchange between the 2-lithio derivative and hexachloroethane. PMID- 15527301 TI - Novel salicylaldehyde-based mineral-supported expeditious synthesis of benzoxazin 2-ones. AB - One-pot montmorillonite K-10 clay supported reactions of either salicylaldehyde/2 hydroxyacetophenone hydrazones and aryl-/alkylureas or salicylaldehydes/2 hydroxyacetophenone and 4-aryl-/alkylsemicarbazides expeditiously yield 3,4 dihydro-4-hydrazino-2H-benz[e]-1,3-oxazin-2-ones (9) via cycloisomerization of the intermediate salicylaldehyde/2-hydroxyacetophenone 4-aryl /alkylsemicarbazones (5) under solvent-free microwave irradiation. Under the same conditions, hydrazines (9) readily underwent reductive dehydrazination on alumina supported copper(II) sulfate to furnish 2H-benz[e]-1,3-oxazin-2-ones (10). PMID- 15527302 TI - One-pot synthesis of cyanuric acid-bridged porphyrin-porphyrin dyads. AB - Stepwise amination of cyanuric chloride (1) with 5-(4-aminophenyl)-10,15,20 triphenylporphyrin (2) and/or its zinc(II) complex (3) enables the synthesis of porphyrin-porphyrin dyads with predetermined free base-free base forms or free base-zinc and zinc-zinc metalation states. Furthermore, the use of aminopropyl silanized silica gel as a scavenger for unwanted byproducts allowed the one-pot synthesis of title porphyrin compounds in high yield and purity with minimum use of preparative column chromatography. PMID- 15527303 TI - Mild and selective palladium-catalyzed dimerization of terminal alkynes to form symmetrical (Z,Z)-1,4-dihalo-1,3-dienes. AB - A regioselective and stereoselective palladium-catalyzed dimerization of terminal alkynes method for the synthesis of symmetrical (Z,Z)-1,4-dihalo-1,3-dienes is presented. In the presence of a catalytic amount of PdX(2) and 3 equiv of CuX(2) (X = Cl and Br), terminal alkynes were dimerized to afford (Z,Z)-1,4-dihalo-1,3 dienes in good yields. The results showed that the effect of solvent had a fundamental influence on the chemoselectivity of the dimerization reaction. The mechanism of the palladium-catalyzed dimerization reaction is also discussed. PMID- 15527304 TI - Gas-phase kinetics and activation parameters for thermal [1,5] hydrogen shifts interconverting monodeuterium- labeled 1,3-cycloheptadienes. AB - The kinetics of thermal equilibrations among monodeuterium-labeled 1,3 cycloheptadienes in the gas phase followed from 154 to 190 degrees C provide activation parameters for the [1,5] shift of a single hydrogen: E(a) = (27.5 +/- 0.9) kcal/mol and log A = 9.7 [corrected] +/- 0.4. These activation parameters imply a comparatively low E(a) barrier balanced by demandingly specific geometric constraints, for DeltaS (170 degrees C) = -17 [corrected] e.u. PMID- 15527305 TI - On the reaction of 1-oxa-4-thiaspiro[4.5]decan-7-one with PhLi. A reinvestigation. AB - Two diastereomeric alcohols are formed from the reaction of 1-oxa-4 thiaspiro[4.5]decan-7-one with PhLi as against the single diastereomer reported earlier. The diastereoselectivity achieved from PhMgBr is significantly higher than with PhLi. The use of Et(2)O/hexane = 2:1 as the solvent for the reaction with PhMgBr offered the highest (14:1) diastereocontrol. PMID- 15527306 TI - Diastereoselective total synthesis of isocarbacyclin from L-ascorbic acid. AB - Diastereoselective total synthesis of isocarbacyclin, which features a fused bicyclic key intermediate available from l-ascorbic acid, is described. The key intermediate was prepared in multigram quantities by the Pauson-Khand reaction of l-ascorbic acid-based (R)-4,4-diallyl-2,2-dimethyl-5-(trimethylsilyl)ethynyl-1,3 dioxolane (3), discriminating diastereotopic groups and faces of the geminal allyl substituents. PMID- 15527307 TI - Palladium-catalyzed cross-coupling reactions of allylic halides and acetates with indium organometallics. AB - The palladium(0)-catalyzed cross-coupling reaction of allylic halides and acetates with indium organometallics is reported. In this synthetic transformation, triorganoindium compounds and tetraorganoindates (aryl, alkenyl, and methyl) react with cinnamyl and geranyl halides and acetates to afford the S(N)2 product regioselectively and in good yield. The reaction proceeds with net inversion of the stereochemical configuration. PMID- 15527308 TI - Synthesis of functional meso-aryl porphomonomethenes and porphodimethenes: application to the preparation of a chiral calix[4]phyrin dimer. AB - Reaction of 5,5-dimethyldipyrromethane (1) with electron-deficient aryl aldehydes in the presence of BF(3)-Et(2)O and NH(4)Cl in propionitrile constitutes efficient, easy access to unprecedented, functional porphomonomethenes together with the expected porphodimethenes (calix[4]phyrins). Alternatively, when the reaction was carried out in CH(2)Cl(2) in the presence of an acid and Florisil, the expected bis-arylcalix[4]phyrin was isolated in 41% yield, while no scrambled macrocycle was detected. After reduction of the nitro function, porphomonomethene 9 was efficiently condensed with the binaphthyl diacyl chloride (10) leading to the first chiral calix[4]phyrin dimer (11) that exhibits a moderate enantiorecognition toward the enantiomers of malic acid. PMID- 15527309 TI - Reactivity of pyrido[4,3,2-kl]acridines: regioselective formation of 6 substituted derivatives. AB - Pyrido[4,3,2-kl]acridines represent a new class of heterocycles, isomers of marine alkaloids. The 7H-pyrido[4,3,2-kl]acridine reacts as an electron rich heterocycle, and in particular via electrophilic substitution such as H/D exchange and the Vilsmeier-Haack reaction. The reaction is fully regioselective and gives the corresponding 6-substituted derivatives. The pyrido[4,3,2 kl]acridin-4-one reacts with amines and thiol, via 1,4-Michael addition to give the 6-amino or 6-thio analogues in a very efficient way. Molecular calculations account for the observed regioselectivity. PMID- 15527310 TI - A new and efficient synthetic method for 15N3-labeled cytosine nucleosides: Dimroth rearrangement of cytidine N3-oxides. AB - The treatment of (15)N(4)-labeled cytidine N(3)-oxide and (15)N(4)-labeled 2' deoxycytidine N(3)-oxide, prepared from the appropriate unprotected uridines in three reaction steps, with benzyl bromide in the presence of excess lithium methoxide allowed the smooth occurrence of their Dimroth rearrangement even under mild conditions leading to the corresponding (15)N(3)-labeled uridine 4-O benzyloximes which can easily undergo the reductive N-O bond cleavage to give the desirable (15)N(3)-labeled cytosine nucleosides in high total yields. PMID- 15527311 TI - Synthesis of N-methylpyrrole and N-methylimidazole amino acids suitable for solid phase synthesis. AB - New and higher yielding synthetic routes to N-protected N-methylpyrrole and N methylimidazole amino acids are introduced to circumvent difficulties associated with established schemes. Key steps in each synthesis include copper-mediated cross-coupling reaction to directly install a carbamate-protected 4-amine in the N-methylpyrrole derivative and effective nitration followed by a one-pot reduction/Boc protection of the amine in the synthesis of the N-Me-imidazole amino acid. PMID- 15527312 TI - Intramolecular ionic Diels-Alder reactions of alpha-acetylenic acetals. AB - The intramolecular ionic Diels-Alder reaction of alpha-acetylenic acetals as a precursor of the propargyl cation has been investigated in the presence of Lewis acids and in protic acids. The reaction of diene-tethered alpha-acetylenic acetals (1-2) with formic acid yielded the regioselective intramolecular ionic Diels-Alder reaction products, bicyclodienal (9) and bicyclodienone (11) derivatives, in good yields. PMID- 15527313 TI - Synthesis of monodentate chiral spiro phosphonites and the electronic effect of ligand in asymmetric hydrogenation. AB - New monodentate chiral phosphonites were synthesized from enantiomerically pure 1,1'-spirobiindane-7,7'-diol. The phosphonites 2 were efficient ligands for the Rh-catalyzed asymmetric hydrogenation of alpha- and beta-dehydroamino acid derivatives, providing the amino acids in high enantioselectivities. The study of electronic effect showed that the electron-withdrawing substitutent on the P phenyl ring of the phosphonite ligand dramatically decreased both the reactivity and enantioselectivity of the ligand. PMID- 15527314 TI - Density functional study of the optical rotation of glucose in aqueous solution. AB - Optical rotation values were calculated for the eight most abundant structures of glucose in aqueous solution, following the TD-DFT/GIAO approach for the property and the PCM description for the solvent. The results show that all alpha structures give a large positive contribution to the OR property, while the beta structures give both positive and negative contributions. The good agreement of the calculated OR, obtained as a Boltzmann average of the property of the eight conformers, with experimental data proves the validity of the quantum-mechanical approach and of the solvent modelization. PMID- 15527315 TI - Organocatalytic asymmetric hydroxylation of beta-keto esters: metal-free synthesis of optically active anti-diols. AB - The organocatalytic alpha-hydroxylation of beta-keto esters using cinchona alkaloid derivatives as the catalyst and peroxides as the terminal oxidant has been investigated and is shown to proceed in high yields and with good enantioselectivity. The scope of the reaction is demonstrated for various substrates, and furthermore, the preparation of optically active anti-diols is presented. PMID- 15527316 TI - Organocatalyzed solvent-free aza-Henry reaction: a breakthrough in the one-pot synthesis of 1,2-diamines. AB - A nitrogen-containing superbase such as TMG was found to be an effective catalyst for the reaction between N-diphenylphosphinoyl imines and nitroalkanes. Exploiting a protocol that avoids the use of any solvent also during workup procedure, we synthesized a series of beta-nitroamines in excellent yields and high diastereomeric ratios. These results, combined with the capability of the indium in conjunction with Zn as the stoichiometric reducing agent to perform in aqueous medium reduction of the nitro group under mild reaction conditions, led us to devise a three-step, one-pot synthesis of a range of 1,2-diamines, making use of environmentally friendly procedures in the various steps. PMID- 15527317 TI - A straightforward synthesis of (-)-phaseolinic acid. AB - A concise approach to (-)-phaseolinic acid starting from commercially available (S)-oct-1-yn-3-ol is disclosed. The key steps are a ring-closing metathesis reaction to prepare a C(2)-symmetrical allylic diol and its desymmetrization to a gamma-butyrolactone by using an Ireland-Claisen rearrangement. The 2S,3S,4S configuration of the levogyre natural product has been confirmed. PMID- 15527321 TI - Proteomic patterns as a diagnostic tool for early-stage cancer: a review of its progress to a clinically relevant tool. AB - The pace of development in novel technologies that promise improvements in the early diagnosis of disease is truly impressive. One such technology at the forefront of this revolution is mass spectrometry. New capabilities in mass spectrometry have provided the means for the development of proteomics, and the race is on to find innovative ways to apply this powerful technology to solving the problems faced in clinical medicine. One area that has garnered much attention over the past few years is the use of mass spectral patterns for cancer diagnostics. The use of these so-called 'proteomic patterns' for disease diagnosis relies fundamentally on the pattern of signals observed within a mass spectrum rather than the more conventional identification and quantitation of a biomarker such as in the case of cancer antigen-125- or prostate-specific antigen. The inherent throughput of proteomic pattern technology enables the analysis of hundreds of clinical samples per day. Currently, there are two primary means by which proteomic patterns can be acquired, surface-enhanced laser desorption/ionization (SELDI) and an electrospray ionization (ESI) method that has been popularized under the name, OvaCheck. In this review, an historical perspective on the development of proteomic patterns for the diagnosis of early stage cancers is described. In addition, a critical assessment of the overall technology is presented with an emphasis on the steps required to enable proteomic pattern analysis to become a viable clinical tool for diagnosing early stage cancers. PMID- 15527322 TI - Feasibility of a cost-effective approach to evaluate short tandem repeat markers suitable for chimerism follow-up. AB - BACKGROUND: Precise chimerism monitoring is important for the prediction of the success of allogeneic bone marrow transplantation (BMT). Most of the current procedures employed for chimerism follow-up with short tandem repeat (STR) markers are either time-consuming, labor-intensive, or use expensive assays, making it burdensome to perform large-scale studies of transplanted patients. AIM: To set-up a simple nonradioactive method to investigate a set of STR markers that could be used in the evaluation of chimerism status after allogeneic BMT. METHOD: Six dinucleotide STRs (D2S123, D5S107, CRTL1, D7S500, D11S1356, and TP53) were analyzed by touchdown (TD)-PCR followed by medium size non-denaturing polyacrylamide gel electrophoresis and silver staining. The sensitivity of the approach was evaluated by dilution competition assays. Peripheral blood samples were taken from a group of 50 healthy Argentinean donors, two transplanted patients, and their respective bone marrow donors. Buccal mucosa samples were also obtained from the BMT recipients. RESULTS: Four markers, D2S123, D7S500, D11S1356, and TP53, presented the highest heterozygosities (0.67-0.88) under our experimental system. A sensitivity of 0.8-1.6% for chimerism detection was consistently found for the different STR. The usefulness of these STR in chimerism analysis was illustrated with the screening of related siblings analyzing two transplanted patients with persistent mixed chimerism, which were previously studied by fluorescence in situ hybridization (FISH). Similar proportions of mixed chimerism were obtained with STR analysis compared with those estimated by FISH. DISCUSSION: To our knowledge, this was the first study of mixed chimerism using TD-PCR to achieve a highly specific STR amplification. This approach allows simple and accurate chimerism quantification because it avoids slippage of Taq polymerase on repeat stretches and prevents the differential amplification of the shorter allele. STR heterozygosities and the high level of sensitivity of this method demonstrated that this approach is not only very informative in this population, but is also rapid (taking less than 14 hours) and cost-efficient. CONCLUSION: The data confirms that this method is a useful tool applicable to routine large-scale STR genotyping and mixed chimerism analysis in low-complexity laboratories worldwide. PMID- 15527323 TI - Detection of MYCN amplification and chromosome 1p36 loss in neuroblastoma by cDNA microarray comparative genomic hybridization. AB - BACKGROUND: In the last decade, microarray technology has been extensively used to evaluate gene expression profiles and genome imbalances. We have developed a microarray-based comparative genomic hybridization (CGH) approach to identify MYCN gene amplification and 1p36 chromosome loss, two markers of tumor aggressiveness in neuroblastoma. AIM: The aim was to use microarray CGH technology to detect the two major prognostic markers for neuroblastoma, MYCN amplification and 1p36 chromosome deletion, in neuroblastoma patients and, therefore, confirm the usefulness of this approach in this cancer. METHODS: DNA was purified from 16 tumors containing at least 90% malignant neuroblasts and collected at the onset of disease. Pooled fluorescent-labeled reference and neuroblastoma tumor genomic DNA was hybridized to epoxide-coated glass slides on laboratory-made complementary DNA microarray. The microarray contained cDNA mapped at the 1p36.33-36.1 chromosomal region and MYCN gene. cDNA from the 2q33 q34 and 12p13 chromosomes was used as a control and Arabidopsis thaliana DNA was spotted to control unspecific hybridization. Fluorescence in situ hybridization analysis was also performed to validate results from the microarray CGH. RESULTS: Both MYCN amplification and 1p36 chromosome deletion were detected by microarray CGH. The sensitivity and specificity for 1p36 loss detection were 66.7% and 90.0%, respectively. The method had a sensitivity of 66.7% and specificity of 90.9% to detect MYCN amplification. DISCUSSION: Our results demonstrated that the microarray CGH can be efficiently applied to study DNA gain and loss of specific chromosome regions. PMID- 15527324 TI - Rapid detection of common CARD15 variants in patients with inflammatory bowel disease. AB - BACKGROUND: Three mutations (R702W, G908R, and 1007fs) within the CARD15 gene have been identified as independent risk factors for the development of Crohn's disease (CD). Virtually all studies investigating the occurrence of these mutations in patients with CD have used separate PCR-based methods to screen patient DNA, here we describe a novel multiplex amplification refractory mutation system (ARMS) assay that allows the simultaneous detection of R702W, G908R, and 1007fs, and a fourth CARD15 variant, P268S, at a fraction of the cost of the pre existing genotyping assays. METHODS: Allele-specific primer sets were designed for each CARD15 variant, optimized separately for annealing temperature and MgCl2 and then multiplexed. The mutant- and wild-type-specific primers were split across two tubes so that each multiplex reaction was internally controlled for amplification failure. An additional primer pair specific to beta2-microglobulin was included as an independent control for DNA quality. The specificity of each primer set was tested using positive controls that had been validated by sequencing, and the robustness of the final ARMS assay was assessed by genotyping 111 Caucasian patients with inflammatory bowel disease (IBD). RESULTS: The specificity of each primer set was confirmed using a sequence validated positive control for each of the four CARD15 variants. Of the 111 DNA samples screened with our ARMS assay, a clear CARD15 genotype was obtained for 109 patients. DISCUSSION AND CONCLUSIONS: Given the potential predictive value of R702W, G980R, and 1007fs, a robust genotyping method for these variants would be of considerable value both in diagnostic and research settings. Our ARMS assay only takes 3-4 hours to perform once DNA has been extracted and requires only 1U of Taq DNA polymerase, making it a rapid, reliable, and cost-effective alternative to current CARD15 genotyping methods. PMID- 15527325 TI - Housekeeping genes as internal standards in cancer research. AB - BACKGROUND: Differences in gene expression are frequently encountered in malignant tissues, and have been intensively studied as they can reflect different experimental or clinical conditions. Quantification of the often subtle changes in messenger RNA content is performed through comparison with the expression of endogenous controls. The appropriate choice of these endogenous controls (e.g. housekeeping genes) is critical for meaningful quantitative RNA analysis. The most important characteristics of housekeeping genes are that they are present in all cells and that their expression levels remain relatively constant in different experimental conditions. However, no single housekeeping gene always manifests stable expression levels under all experimental conditions. Therefore, it is necessary to characterize the suitability of various housekeeping genes to serve as internal RNA controls under particular experimental conditions where transcription effects are being tested. AIM: It was the aim of this study to determine the validity of a number of housekeeping genes for their use as internal standards in cancer research. METHODS: The expression of the housekeeping genes porphobilinogen deaminase (PBGD) and mitochondrial ATP synthase 6 (mATPsy6), were compared with the expression of the more commonly used glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We examined a number of cell lines and tumor versus matched normal tissue samples using real-time quantitative (RTq)-PCR. RESULTS: Our findings suggest that in cell lines, all three of the studied housekeeping genes can be used as an internal control. When comparing tumor tissue samples with matched normal tissue samples, we validated mitochondrial ATPsy6 (mATPsy6) as the best choice for a housekeeping gene. CONCLUSION: Since gene expression studies are becoming increasingly important in the clinical environment, especially in cancer diagnosis and treatment, the use of an reliable housekeeping gene in these studies to normalize gene expression is essential. We conclude that a bad choice of housekeeping gene may lead to errors when interpreting experiments involving quantitation of gene expression. Our study demonstrated the usefulness of mATPsy6 as an endogenous control when comparing tumor tissue samples with normal tissue samples. PMID- 15527326 TI - Detection of germline mosaicism in two Duchenne muscular dystrophy families using polymorphic dinucleotide (CA)n repeat loci within the dystrophin gene. AB - BACKGROUND: Approximately one-third of new cases of Duchenne muscular dystrophy (DMD) can be attributed to sporadically arising new mutations, however in the majority of cases the DMD mutation has been inherited from the mother. These female carriers can have either a constitutive or mosaic mutation. AIM: The aim of this study was to determine the segregation of the at-risk haplotype and to find a deletion in the dystrophin gene of patients. METHOD: We analyzed individuals from two families with a history of DMD in order to predict the carrier status of related females. In one of these cases the mother had two affected sons, while in the other one son and two grandchildren were affected; therefore we predict that the mother would be an obligatory carrier. RESULTS: Haplotype analysis of the DMD loci revealed that in the two families both the healthy and affected brothers had inherited the same X maternal chromosome. However, the affected brother carried a deletion, which was absent in the unaffected sibling. CONCLUSION: These findings suggested that the mothers in the two families were germline mosaics for the DMD gene. The results of this study demonstrate the usefulness of the methodology that combine the haplotype analysis with the identification of the mutation in order to detect hidden germline mosaicisms and, thus, improve genetic counseling. PMID- 15527327 TI - Use of TP53 reference materials to validate mutations in clinical tissue specimens by single-strand conformational polymorphism analysis. AB - BACKGROUND: As genetic information moves from basic research laboratories in to the clinical testing environment, there is a critical need for reliable reference materials for the quality assurance of genetic tests. A panel of 12 plasmid clones containing wild-type or point mutations within exons 5-9 have been developed as reference materials for the detection of TP53 mutations. AIM: The goal of this study was to validate the reference materials in providing quality assurance for the detection of TP53 mutations in clinical specimens. METHODS: We studied 33 gynecological samples, 11 apparently normal samples and 22 malignant tumors of various origins. Mutations were identified using single-strand conformational polymorphism analysis with both slab gel and capillary electrophoresis. All DNA samples were amplified with fluorescently labeled PCR primers specific for exons 5-9 for mutation detection. RESULTS: Of the 33 patient samples tested, mutations and polymorphisms were found in six specimens in three of the five exons scanned; no mutations were found in exons 7 or 9. Both a mutation and polymorphism were found in non-malignant specimens from the control group. The mutations were confirmed by DNA sequence analysis of the regions scanned. CONCLUSIONS: Mutations and polymorphisms were detected in the clinical samples. All of the mutations were silent except for one non-conservative mutation in exon 5, codon 181. This study demonstrates the usefulness of the National Institute of Standards and Technology (NIST) TP53 reference panel in TP53 mutation detection in clinical tissue specimens. PMID- 15527328 TI - Detection of chromosomal- and plasmid-encoded virulence-associated epidemiological markers in Yersinia enterocolitica strains isolated from clinical cases: a comparative study. PMID- 15527329 TI - Enhanced infrared absorption spectra of self-assembled alkanethiol monolayers using the extraordinary infrared transmission of metallic arrays of subwavelength apertures. AB - The surface-plasmon-mediated, extraordinary transmission of metallic arrays of subwavelength apertures has been used as the light source for absorption studies of self-assembled monolayers on metal. Enhanced infrared absorption spectra of a sequence of alkanethiol self-assembled monolayers on copper were recorded for carbon chain lengths varying from 8 to 18 atoms. Transition positions and intensities are presented over a large range of the infrared region. The connection between the vibrational modes of the CH(2) wagging progression and the infinite methylene chain is explored using a traditional coupled oscillator approach and a new cluster perspective. PMID- 15527330 TI - Hydrophobic hydration of alkanes: its implication for the property of amorphous solid water. AB - We measured the incorporation of adsorbed alkanes in and their desorption from the amorphous solid water (ASW) by means of secondary ion mass spectroscopy and temperature programmed desorption. The heavier alkanes such as butane and hexane are incorporated completely in the bulk of the nonporous ASW layer below 100 K probably due to the preferential formation of ice structures around the solute molecules. The self-diffusion of water molecules occurs above the glass transition temperature (136 K). The liquid water emerges above 165 K, as evidenced by simultaneous occurrence of the dehydration of alkanes and the morphological change of the water layer induced by the surface tension. PMID- 15527331 TI - On the electron tunneling in molecules: a generalized orthogonalization procedure for finding tunneling orbitals. AB - A simple method for finding tunneling orbitals of a long-distance electron transfer system is described. The procedure is an approximate biorthogonalization of many-electron donor and acceptor states in which the corresponding orbitals of the system are obtained; one pair of such corresponding orbitals describes the tunneling electron. Usually, the biorthogonalization is performed on the states that describe simultaneously both donor and acceptor complexes and the bridge connecting them, which make the procedure computationally demanding. In this paper a significant simplification of the procedure is proposed. In the procedure, to identify the tunneling orbitals roughly one half of the system is needed--only donor complex, or acceptor complex, plus part of the bridge. The procedure is practically equivalent to biorthogonalization, but computationally much simpler. Applications are illustrated with the results for a model of biological long-distance electron transfer in Ru-modified azurin, and electron transfer involving heme a of cytochrome c oxidase. PMID- 15527332 TI - An analysis of core effects on shape-consistent pseudopotentials. AB - Large core (seven-valence electrons) shape-consistent averaged relativistic pseudopotentials (AREP) including core effects have been derived for the halogen series (Cl,Br,I,At). The influence of core effects on the spin-orbit splitting of the halogen and alkali atoms is clearly demonstrated within an all-electron four component atomic reference calculation by means of a perturbation analysis. In particular, it is shown that AREPs extracted at the Dirac-Coulomb-Fock level, which already include spin-orbit polarization effects, give excellent results for atomic spectroscopy and equilibrium distances of halogen dimers. We also show that in our approach the core effects, included by configuration interaction using the numerical GRASP code, are transferred to the averaged orbital one electron energy, defined in a perturbational way. This leads to a modification of the extracted AREPs by core effects, which is illustrated by calculations of the first atomic excited states using these AREPs. These results support the validity of including core effects directly in the AREPs extracted in a shape-consistent scheme. The transferability to the atomic excited states as well as to the molecular case is also verified. PMID- 15527333 TI - Feature activated molecular dynamics: an efficient approach for atomistic simulation of solid-state aggregation phenomena. AB - An efficient approach is presented for performing efficient molecular dynamics simulations of solute aggregation in crystalline solids. The method dynamically divides the total simulation space into "active" regions centered about each minority species, in which regular molecular dynamics is performed. The number, size, and shape of these regions is updated periodically based on the distribution of solute atoms within the overall simulation cell. The remainder of the system is essentially static except for periodic rescaling of the entire simulation cell in order to balance the pressure between the isolated molecular dynamics regions. The method is shown to be accurate and robust for the Environment-Dependant Interatomic Potential (EDIP) for silicon and an Embedded Atom Method potential (EAM) for copper. Several tests are performed beginning with the diffusion of a single vacancy all the way to large-scale simulations of vacancy clustering. In both material systems, the predicted evolutions agree closely with the results of standard molecular dynamics simulations. Computationally, the method is demonstrated to scale almost linearly with the concentration of solute atoms, but is essentially independent of the total system size. This scaling behavior allows for the full dynamical simulation of aggregation under conditions that are more experimentally realizable than would be possible with standard molecular dynamics. PMID- 15527334 TI - Modified quantum trajectory dynamics using a mixed wave function representation. AB - Dynamics of quantum trajectories provides an efficient framework for description of various quantum effects in large systems, but it is unstable near the wave function density nodes where the quantum potential becomes singular. A mixed coordinate space/polar representation of the wave function is used to circumvent this problem. The resulting modified trajectory dynamics associated with the polar representation is nonsingular and smooth. The interference structure and the nodes of the wave function density are described, in principle, exactly in the coordinate representation. The approximate version of this approach is consistent with the semiclassical linearized quantum force method [S. Garashchuk and V. A. Rassolov, J. Chem. Phys. 120, 1181 (2004)]. This approach is exact for general wave functions with the density nodes in a locally quadratic potential. PMID- 15527335 TI - Reducing a chemical master equation by invariant manifold methods. AB - We study methods for reducing chemical master equations using the Michaelis Menten mechanism as an example. The master equation consists of a set of linear ordinary differential equations whose variables are probabilities that the realizable states exist. For a master equation with s(0) initial substrate molecules and e(0) initial enzyme molecules, the manifold can be parametrized by s(0) of the probability variables. Fraser's functional iteration method is found to be difficult to use for master equations of high dimension. Building on the insights gained from Fraser's method, techniques are developed to produce s(0) dimensional manifolds of larger systems directly from the eigenvectors. We also develop a simple, but surprisingly effective way to generate initial conditions for the reduced models. PMID- 15527336 TI - Time-dependent exchange-correlation current density functionals with memory. AB - Most present applications of time-dependent density functional theory use adiabatic functionals, i.e., the effective potential at time t is determined solely by the density at the same time. This paper discusses a method that aims to go beyond this approximation, by incorporating "memory" effects: the derived exchange-correlation potential will depend not only on present densities but also on the past. In order to ensure the potentials are causal, we formulate the action on the Keldysh contour for electrons in electromagnetic fields, from which we derive suitable Kohn-Sham equations. The exchange-correlation action is now a functional of the electron density and velocity field. A specific action functional is constructed which is Galilean invariant and yields a causal exchange-correlation vector potential for the Kohn-Sham equations incorporating memory effects. We show explicitly that the net exchange-correlation Lorentz force is zero. The potential is consistent with known dynamical properties of the homogeneous electron gas (in the linear response limit). PMID- 15527337 TI - Model for small-sample bias of free-energy calculations applied to Gaussian distributed nonequilibrium work measurements. AB - We present a model for the bias of free-energy differences when determined using the nonequilibrium work (NEW) formalism due to Jarzynski. Input to the model is the distribution of work values underlying the NEW calculation, and the bias is estimated by assuming that all of the inaccuracy is incurred by failure to sample work values beyond a specific point in the tail of the distribution. The model is formulated considering both small- and large-sample NEW calculations. It is then applied to the study of bias for cases in which the work distribution is Gaussian. The model is shown to give an excellent description of the bias in situations where the bias is a more significant source of error than the sample variance. A scaling law is presented and it is shown that the bias as a function of sampling can be reduced to a single universal curve, approximately valid for all Gaussian work distributions. This result is used to formulate a simple criterion that can be applied to test if a NEW calculation is effectively free of bias. The criterion is shown to be effective even though it uses the measured (and perhaps biased) free energy as an input. PMID- 15527338 TI - Coherent population transfer in molecules coupled with a dissipative environment by intense ultrashort chirped pulse. II. A simple model. AB - We have developed a simple and physically clear picture of adiabatic rapid passage (ARP) in molecules in solution by careful examination of all the conditions needed for ARP. The relaxation effects were considered in the framework of the Landau-Zener model for random crossing of levels. The model enables us to include into consideration non-Markovian Gaussian-correlated noise. It explains all the numerical results obtained in the first paper of the series [B. D. Fainberg and V. A. Gorbunov, J. Chem. Phys. 117, 7222 (2002)], in particular, that for positive chirp pulse excitation relaxation favors more efficient population transfer with respect to the relaxation-free system with frozen nuclear motion. We also relate parameters of non-Markovian Gaussian correlated noise with irreversible dephasing time of an optical transition by calculating the photon echo signal attenuation. PMID- 15527339 TI - Well-balanced basis sets for second-order Moller-Plesset treatment of argon aromatic molecule complexes. AB - Efficient ab initio method for studies of van der Waals complexes of argon and aromatic molecules is presented. It is based on the supermolecular second-order Moller-Plesset (MP2) perturbation theory combined with well-balanced basis sets. The error resulting from incompleteness of such basis sets is almost exactly canceled by the correlation error inherent in the MP2 method. Two basis sets adapted to the MP2 method are selected from various medium-sized basis sets. The standard augmented correlation consistent polarized valence double-zeta basis set and a smaller reduced version derived from it are shown to perform exceptionally well. They are employed in a large scale computation of the potential energy surfaces of argon-benzene and argon-fluorobenzene complexes. The results are critically compared with ab initio high level coupled-cluster calculations and experimental data available. The calculated MP2 equilibrium geometry, dissociation energy, and the vibrational states of the stretching mode are proved to be in excellent agreement with the experiment. However, the bending fundamentals are systematically overestimated by about 1 cm(-1). This deficiency is removed by introducing a simple correction function which improves the MP2 potential energy surface. This function can be easily determined and applied to arbitrary argon-aromatic molecule complexes. The MP2 method is compared to the density-functional theory. Local, semilocal, and hybrid models are tested and the results obtained clearly show that none of these models is capable of accurate description of the van der Waals interaction. PMID- 15527340 TI - Collision photography: polarization imaging of atom-molecule collisions. AB - We report differential scattering experiments on the laser excitation of Na + M collision pairs with M = N(2), CO, C(2)H(2), and CO(2). The collision event is probed by the laser polarization revealing geometric and electronic properties of the collision pair. The experimental data are compared to the results of a Monte Carlo trajectory simulation using ab initio quantum chemical data. PMID- 15527341 TI - Characteristic of structures and pi-hydrogen bond of dimers C2H4-nFn-HF (n=0,1,2). AB - By the counterpoise-correlated potential energy surface method (interaction energy optimization), five structures of the C(2)H(4-n)F(n)-HF (n = 0,1,2) dimers with all real frequencies have been obtained at MP2/aug-cc-pVDZ level. The influence of F substituent effect on the structure and pi-hydrogen bond of dimer has been discussed. For C(2)H(4-n)F(n)-HF (n = 1,2), the pi-hydrogen bonds are elongated comparing with that for C(2)H(4)-HF. For C(2)H(3)F-HF, g-C(2)H(2)F(2) HF, cis-C(2)H(2)F(2)-HF, the pi-hydrogen bonds are further deformed. These changes (elongate, shift, and deformation) of pi-hydrogen bond mainly come from deformation of pi-electron cloud of C=C bond. The pi-electron cloud is pushed towards the one C atom, the pi H-bond shift also to the C direction. Since the two lobes of pi-electron cloud have deviated slightly from the molecular vertical plane passing through C=C bond, the pi-hydrogen bond is sloped. Intermolecular interaction energies of the dimers are calculated to be -3.9 for C(2)H(4)-HF, 2.8 for C(2)H(3)F-HF, -2.1 for g-C(2)H(2)F(2)-HF, -1.6 for cis-C(2)H(2)F(2)-HF, 1.3 kcal/mol for trans-C(2)H(2)F(2)-HF, at CCSD(T)/aug-cc-pVDZ level. PMID- 15527342 TI - The one-particle Green's function method in the Dirac-Hartree-Fock framework. II. Third-order valence ionization energies of the noble gases, CO and ICN. AB - In this paper we present the third-order extension of the four-component one particle propagator method in the non-Dyson version of the algebraic diagrammatic construction (ADC) for the calculation of valence ionization energies. Relativistic and electron correlation effects are incorporated consistently by starting from the Dirac-Hamiltonian. The ADC equations derived from the Feynman diagrams can hereby be used in their spin-orbital form and need not be transformed to the spin-free version as required for a nonrelativistic treatment. For the calculation of the constant self-energy contribution the Dyson expansion method was implemented being superior to a perturbational treatment of sigma(infinity). The Dirac-Hartree-Fock- (DHF-) ADC(3) was applied to the calculation of valence photoionization spectra of the noble gas atoms, carbon monoxide and ICN now also reproducing spin-orbit features in the spectrum. Comparison with DHF-ADC(2), nonrelativistic ADC(3), and experimental data was made in order to demonstrate the characteristics and performance of the method. PMID- 15527343 TI - Molecular elimination in photolysis of fluorobenzene at 193 nm: internal energy of HF determined with time-resolved Fourier-transform spectroscopy. AB - Following photodissociation of fluorobenzene (C(6)H(5)F) at 193 nm, rotationally resolved emission spectra of HF(1C2H4 + HF product energy partitioning. AB - Direct dynamics classical trajectory simulations were performed to study product energy partitioning in C(2)H(5)F-->C(2)H(4)+HF dissociation. The intrinsic reaction coordinate potential energy curve, reaction energetics, and transition state (TS) properties were calculated for this reaction at different levels of electronic structure theory, and MP2/6-31G( *) was chosen as a meaningful and practical method for performing the direct dynamics. The trajectories show that the HF bond, uncoupled from the other degrees of freedom, is formed within the first 10 fs as the system moves from the TS towards products. The populations of the HF vibration states, determined from the simulations, decrease monotonically as found from experiments. However, the simulation's populations for the low and high energy vibration states are larger and smaller, respectively, than the experimental results. The HF rotational temperature found from the simulations is in agreement with experiment. Increasing the TS's excess energy gives higher rotational temperatures for both C(2)H(4) and HF. Energy is partitioned to the products from both the excess energy in the TS and the potential energy release in the exit channel. Partitioning from these two energy sources is distinguished by varying the TS's excess energy. An analysis of the simulation's energy disposal shows that the fractions of the excess energy partitioned to relative translation, C(2)H(4) vibration, C(2)H(4) rotation, HF vibration, and HF rotation, are 0.17, 0.64, 0.076, 0.067, and 0.046, respectively, and are in good agreement with previous simulations on empirical potentials and experiments. The partitioning found for the potential energy release is 81%, <0.05%, 5%, 11%, and 3% to relative translation, C(2)H(4) vibration, C(2)H(4) rotation, HF vibration, and HF rotation. This result is substantially different than the deduction from experiments, which summarizes the partitioning as 20%, 45%, 24%, and <12% to relative translation, C(2)H(4) vibration+rotation, HF vibration, and HF rotation. Possible origins of the difference between the simulations and experiments in the release of the potential energy is discussed. PMID- 15527347 TI - Photoelectron kinetic energy dependence in near threshold ionization of NO from A state studied by time-resolved photoelectron imaging. AB - Photoelectron angular distributions in the laboratory frame (LF-PADs) from the A((2)sigma(+)) state of NO molecule were measured by femtosecond time-resolved photoelectron imaging with (1 + 1(')) resonance enhanced multiphoton ionization via the A state. High-precision measurements of the anisotropy parameters of LF PADs were performed for the photoelectron kinetic energy from 0.03 to 1.05 eV as a function of the pump-probe delay time. The revival feature of the rotational wave packet on the A state was clearly observed in the time dependence of the photoelectron anisotropy parameters. By approximating the phase shifts of the photoelectron partial waves by the quantum defects in the high-lying Rydberg states using the multichannel quantum defect theory, the energy-dependent photoionization transition dipole moments were determined, for the first time, from time-dependent LF-PADs measured by time-resolved photoelectron spectroscopy. PMID- 15527348 TI - Evaluation of canonical and microcanonical nonadiabatic reaction rate constants by using the Zhu-Nakamura formulas. AB - We consider a problem of calculating both thermal and microcanonical rate constants for nonadiabatic chemical reactions. Instead of using the conventional transition state theory, we use a generalized seam surface and introduce a concept of a coordinate dependent effective nonadiabatic transition probability based on the Zhu-Nakamura theory which can treat the nonadiabatic tunneling properly. The present approach can be combined with Monte Carlo method so as to be applicable to chemical reactions in complicated systems. The method is demonstrated to work well in wide energy and temperature range. Numerical tests also show that it is very essential for accurate evaluation of the thermal rate constant to use the generalized seam surface and take into account the nonadiabatic tunneling effect. PMID- 15527349 TI - Potential energy surface for H2O((3)A(")) from accurate ab initio data with inclusion of long-range interactions. AB - A new potential energy surface for the O((3)P) + H(2) system in the lowest (3)A(") state is built using ab initio data calculated by Rogers et al. [J. Phys. Chem. A 104, 2308 (2000)] and the double many-body expansion formalism. It incorporates a semiempirical model of long-range interactions, which should play an important role at low collision energies. Preliminary quasiclassical trajectory results at 12.6 kcal/mol collision energy, show that the deeper van der Waals region described in this new surface translates into a four times higher cross section than that of Rogers' (3)A(") surface. To confirm this hypothesis, a second surface was calibrated. The two surfaces are fitted with rmsd<0.5 kcal/mol and differ mainly on the depth of the van der Waals region. That difference in the van der Waals region corresponds to a 22% lower cross section of the less deep surface, which is still three times higher than the equivalent results from Rogers' (3)A(") surface. This study reflects the importance of a correct description of van der Waals forces on potential energy surfaces. PMID- 15527350 TI - Polarization quantum beat spectroscopy of HCF(A1A"). I. 19F and 1H hyperfine structure and Zeeman effect. AB - To further investigate the (19)F and (1)H nuclear hyperfine structure and Zeeman effect in the simplest singlet carbene, HCF, we recorded polarization quantum beat spectra (QBS) of the pure bending levels 2(0) (n) with n = 0-7 and combination bands 1(0) (1)2(0) (n) with n = 1-6 and 2(0) (n)3(0) (1) with n = 0-3 in the HCF A(1)A(")<--X(1)A(') system. The spectra were measured under jet-cooled conditions using a pulsed discharge source, both at zero field and under application of a weak magnetic field (<30 G). Analysis yielded the nuclear spin rotation constants C(aa) and weak field Lande g(aa) factors. Consistent with a two-state model, the majority of observed vibrational levels exhibit a linear correlation of C(aa) and g(aa), and our analysis yielded effective (a) hyperfine constants for the (19)F and (1)H nuclei (in MHz) of 728(23) and 55(2), respectively. The latter was determined here owing to the high resolving power of QBS. The vibrational state selectivity of the (19)F hyperfine constants is discussed, and we suggest that the underlying Renner-Teller interaction may play an important role. PMID- 15527351 TI - Polarization quantum beat spectroscopy of HCF(A1A"). II. Renner-Teller and spin orbit mixing in the simplest singlet carbene. AB - To further investigate the Renner-Teller (RT) effect and spin-orbit mixing in the A(1)A(")<--X(1)A(') system of the simplest singlet carbene, HCF, we report a detailed analysis of the K(a) = 1<--0 subband of 2(0) (4) using polarization quantum beat spectroscopy in combination with fluorescence excitation spectroscopy and lifetime measurements. This subband is perturbed both by RT and spin-orbit interactions, which are clearly differentiated due to the order-of magnitude difference in matrix elements. We show that RT induced mixing with a high vibrational level of X(1)A(') leads to a splitting of this subband, and while the higher energy member is rotationally unperturbed, every line in the lower energy member is perturbed by spin-orbit mixing with background levels of a(3)A("), as evidenced by large (19)F and (1)H hyperfine constants and Lande g factors. In contrast, the higher energy subband exhibits very small Lande g factors and hyperfine constants, which is explained within a model that incorporates only the A(1)A(")-X(1)A(') interaction. We thus demonstrate that polarization quantum beat spectra provides efficient discrimination between RT and spin-orbit interactions. Analysis of the lower energy subband in concert with ab initio electronic structure calculations has yielded the first information on the (19)F and (1)H hyperfine structure of the a(3)A(") state and the magnitude of the spin-orbit matrix elements. PMID- 15527352 TI - Spectroscopy of Cs attached to helium nanodroplets. AB - Cesium oligomers are formed on helium nanodroplets which are doped with one or a few Cs atoms. The monomer absorption of the first electronic p<--s transition upon laser excitation is probed. Spectra employing laser-induced fluorescence, beam depletion, and resonant photoionization are compared. In particular, mass resolved photoionization allows us to specifically probe excitation induced processes such as, e.g., the formation of cesium-helium exciplexes. Absorption spectra of Cs dimers and trimers are recorded in the spectral region accessible by a Ti:sapphire laser. Assignment of dimer spectra is achieved by comparison with model calculations based on ab initio potentials. Electronic absorption lines of Cs trimers are attributed to transitions in the quartet manifold. PMID- 15527353 TI - Ultrafast vibrational spectroscopy of water and aqueous N-methylacetamide: Comparison of different electronic structure/molecular dynamics approaches. AB - Kwac and Cho [J. Chem. Phys. 119, 2247 (2003)] have recently developed a combined electronic structure/molecular dynamics approach to vibrational spectroscopy in liquids. The method involves fitting ab initio vibrational frequencies for a solute in a cluster of solvent molecules to a linear combination of the electrostatic potentials on the solute atoms due to the charges on the solvent molecules. These authors applied their method to the N-methylacetamide-D/D(2)O system. We (S. A. Corcelli, C. P. Lawrence, and J. L. Skinner, [J. Chem. Phys. 120, 8107 (2004)]) have recently explored a closely related method, where instead of the electrostatic potential, the solute vibrational frequencies are fit to the components of the electric fields on the solute atoms due to the solvent molecules. We applied our method to the HOD/D(2)O and HOD/H(2)O systems. In order to make a direct comparison of these two approaches, in this paper we apply their method to the water system, and our method to the N-methylacetamide system. For the water system we find that the electric field method is superior to the potential approach, as judged by comparison with experiments for the absorption line shape. For the N-methylacetamide system the two methods are comparable. PMID- 15527355 TI - Physics of single molecule fluctuations in surface enhanced Raman spectroscopy active liquids. AB - Surface enhanced Raman spectroscopy (SERS) of dyes in solution allows the study of the differences between ensemble averaged spectroscopic signals and single molecular events. We address several outstanding issues in single molecule detection via SERS; in particular, evidence for single molecule vibrational pumping and/or single molecule laser heating, the statistics of hotspots in the liquid, and anti-Stokes/Stokes anomalies. We demonstrate that anti-Stokes/Stokes ratios are a very unreliable measure of temperature, because the two processes are affected differently by the underlying frequency-dependent plasmon resonances. Subtle hints of vibrational pumping and/or heating in single molecules can only obtained via careful cross correlations between the parameters (frequency position, width, and intensity) of the Stokes signals for different excitation lasers. We introduce the use of single-peak parameter cross correlations for the study of these phenomena. PMID- 15527354 TI - Spectral diffusion in a fluctuating charge model of water. AB - We apply the combined electronic structure/molecular dynamics approach of Corcelli, Lawrence, and Skinner [J. Chem. Phys. 120, 8107 (2004)] to the fluctuating charge (SPC-FQ) model of liquid water developed by Rick, Stuart, and Berne [J. Chem. Phys. 101, 6141 (1994)]. For HOD in H(2)O the time scale for the long-time decay of the OD stretch frequency time-correlation function, which corresponds to the time scale for hydrogen-bond rearrangement in the liquid, is about 1.5 ps. This result is significantly longer than the 0.9 ps decay previously calculated for the nonpolarizable SPC/E water model. Our results for the SPC-FQ model are in better agreement with recent vibrational echo experiments. PMID- 15527356 TI - Fragility by elastic incoherent neutron scattering. AB - The present work furnishes an operative definition for the fragility degree by using elastic incoherent neutron scattering (EINS). Such a definition is based on the relation between viscosity, a macroscopic quantity, and the atomic mean square displacement, which refers to a nanoscopic property. This procedure has been used to analyze a set of glass-forming systems and it allows to obtain a linear dependence of the fragility parameter M, obtained by EINS, on the fragility parameter m, obtained by viscosity measurements. PMID- 15527357 TI - Crossover model for the work of critical cluster formation in nucleation theory. AB - We propose a relation for the work of critical cluster formation in nucleation theory W for the systems with long-range interparticle interactions. The method of bridge functions is used to combine the system behavior at sufficiently small quenches, adequately predicted by the classical nucleation theory, with nonclassical effects at deep quenches in the vicinity of the thermodynamic spinodal, described within the framework of the field theoretical approach with an appropriate Ginzburg-Landau functional. The crossover between the two types of nucleation behavior takes place in the vicinity of the kinetic spinodal where the lifetime of a metastable state is of the order of the relaxation time to local equilibrium. We argue that the kinetic spinodal corresponds to the minimum of the excess number of molecules in the critical cluster. This conjecture leads to the form of W containing no adjustable parameters. The barrier scaling function Gamma = W/W(cl), where W(cl) is the classical nucleation barrier, depends parametrically on temperature through the dimensionless combination of material properties. The results for argon nucleation are presented. PMID- 15527358 TI - Kinetics and mechanisms of crystal growth and diffusion in a glass-forming liquid. AB - Extensive data on the viscosity, covering 15 orders of magnitude, and crystal growth rate, covering seven orders of magnitude, of liquid diopside (CaO.MgO.2SiO(2)) were collected in a wide range of undercoolings from 1.10T(g) to 0.99T(m) (T(g) is the glass transition temperature and T(m) the melting point). The raw growth rate data were corrected for the increased interfacial temperature produced by the heat released during crystallization. A detailed analysis confirms that growth mediated by screw dislocations reasonably explain the experimental data in these wide ranges of temperatures and growth rates. Effective diffusion coefficients were calculated from crystal growth rates and from viscosity, and were then compared with measured self-diffusion coefficients of silicon and oxygen in diopside melt. The results show that oxygen and silicon control the diffusion dynamics involved in crystal growth and viscous flow. This study not only unveils the transport mechanism in this complex liquid, but also validates the use of viscosity (through the Stokes-Einstein or the Eyring equations) to account for the kinetic term of the crystal growth expression in a wide range of temperatures. PMID- 15527359 TI - Phase transition dynamics of liquid phase precipitation from a supersaturated gas mixture. AB - This work presents a self-consistent description of phase transition dynamics of disperse liquid phase precipitating from a supersaturated gas mixture. The unified approach integrates the macroscale transport phenomena of cloud dynamics with the essential microphysical kinetic processes of droplet condensation, evaporation, and droplet collisions simultaneously taking place in stochastic population of liquid droplets. A complete set of governing equations with well defined dissipative fluxes and kinetic rates is derived for phase transition dynamics from nucleation to postnucleation to coarsening stages. The local thermodynamics of precipitating system, which is considered as ternary mixture of disperse liquid phase and water vapor with dry air, is redefined to explicitly include on equal basis both the vapor content and liquid content into the fundamental thermodynamic relations and equation of state. The molecular kinetic flux regularization method for growth of submicron droplets is reexamined to include, among others, significant contribution of vapor molecular energy flux into total heat flux, resulting in new expressions for the droplet temperature, growth rate, and effective diffusion coefficients. The local kinetic rates are determined on the basis of microscale kinetic equation for the droplet distribution function. This is in contrast to commonly used semiempirical parametrization schemes for kinetic rates with adjustable parameters, wherein the probabilistic aspects of microphysical processes are not rigorously addressed. Stochastic diffusion interactions among droplets competing for the available water vapor and modifications in the kinetic equation for droplets growing in stochastic population with direct long-range diffusion interactions amongst them are discussed and formulated as well. PMID- 15527360 TI - Constant-volume heat capacity in a near-critical fluid from Monte Carlo simulations. AB - We consider a near-critical fluid of hard spheres with short-range interactions (approximately r(-6)) and obtain its constant-volume heat capacity C(V) by means of Monte Carlo calculations in the canonical ensemble. The question addressed is whether or not the heat capacities of the finite-size systems studied in simulations can provide a reliable indication of nonclassical criticality. For the model fluid considered here this is found to be the case. The heat capacity along the critical isochore shows a peak near the critical temperature, with a system size dependence that is consistent with the known Ising universality class of the model. The relevance of our results to recent attempts to determine the universality class of ionic fluids through calculations of C(V) is briefly discussed. PMID- 15527361 TI - Dynamics of glass-forming liquids. VIII. Dielectric signature of probe rotation and bulk dynamics in branched alkanes. AB - We have measured the dielectric relaxation of several glass forming branched alkanes with very low dielectric loss in the frequency range 50 Hz-20 kHz. The molecular liquids of this study are 3-methylpentane, 3-methylheptane, 4 methylheptane, 2,3-dimethylpentane, and 2,4,6-trimethylheptane. All liquids display asymmetric loss peaks typical of supercooled liquids and slow beta relaxations of similar amplitudes. As an unusual feature, deliberate doping with 2-ethyl-1-hexanol, 5-methyl-2-hexanol, 2-methyl-1-butanol, 1-propanol, or 2 methyltetrahydrofuran at the 1 wt % level generates additional relaxation peaks at frequencies below those of the alpha relaxation. The relaxation times of these sub-alpha-peaks increase systematically with the size of the dopant molecules. Because these features are spectrally separate from the bulk dynamics, the rotational behavior and effective dipole moments of the probes can be studied in detail. For the alcohol guest molecules, the large relative rotational time scales and small effective dipole moments are indicative of hydrogen bonded clusters instead of individual molecules. PMID- 15527362 TI - Three-variable reversible Gray-Scott model. AB - Even though the field of nonequilibrium thermodynamics has been popular and its importance has been suggested by Demirel and Sandler [J. Phys. Chem. B 108, 31 (2004)], there are only a few investigations of reaction-diffusion systems from the aspect of thermodynamics. A possible reason is that model equations are complicated and difficult to analyze because the corresponding chemical reactions need to be reversible for thermodynamical calculations. Here, we introduce a simple model for calculation of entropy production rate: a three-variable reversible Gray-Scott model. The rate of entropy production in self-replicating pattern formation is calculated, and the results are compared with those reported based on the Brusselator model in the context of biological cell division. PMID- 15527363 TI - Structural and electronic evolution of the As(OH)3 molecule in high temperature aqueous solutions: an x-ray absorption investigation. AB - The geometrical and electronic structure of the arsenious acid molecule As(OH)(3) in aqueous solutions has been investigated by x-ray absorption spectroscopy (XAS) within extended x-ray absorption spectroscopy (EXAFS) and x-ray absorption near edge structure (XANES), using realistic first-principle calculations in the latter case. This investigation was performed on aqueous solutions of arsenious acid from ambient to supercritical conditions (P = 250 and 600 bars, T 34 degrees C). This means PNiPAM molecules move to the cold side for temperatures T < 34 degrees C, whereas they move to the warm side for T > 34 degrees C. This is the first nonaqueous polymeric system for which a sign change with temperature has been observed. We performed static and dynamic light scattering experiments in the same temperature range. The second virial coefficient determined from dilute solutions by static light scattering (SLS) was positive in the comparable temperature range. The results of SLS for the semidilute solution showed a strong repulsion among PNiPAM chains which was enhanced by increasing temperature. These results imply that the observed thermally induced sign change of S(T) does not depend on the intermolecular interactions among PNiPAM chains. PMID- 15527383 TI - The Kirkwood-Buff theory and the effect of cosolvents on biochemical reactions. AB - Cosolvents added to aqueous solutions of biomolecules profoundly affect protein stability, as well as biochemical equilibria. Some cosolvents, such as urea and guanidine hydrochloride, denature proteins, whereas others, such as osmolytes and crowders, stabilize the native structures of proteins. The way cosolvents interact with biomolecules is crucial information required to understand the cosolvent effect at a molecular level. We present a statistical mechanical framework based upon Kirkwood-Buff theory, which enables one to extract this picture from experimental data. The combination of two experimental results, namely, the cosolvent-induced equilibrium shift and the partial molar volume change upon the reaction, supplimented by the structural change, is shown to yield the number of water and cosolvent molecules bound or released during a reaction. Previously, denaturation experiments (e.g., m-value analysis) were analyzed by empirical and stoichiometric solvent-binding models, while the effects of osmolytes and crowders were analyzed by the approximate molecular crowding approach for low cosolvent concentration. Here we synthesize these previous approaches in a rigorous statistical mechanical treatment, which is applicable at any cosolvent concentration. The usefulness and accuracy of previous approaches was also evaluated. PMID- 15527384 TI - Coarse-grained model for phospholipid/cholesterol bilayer. AB - We construct a coarse-grained (CG) model for dipalmitoylphosphatidylcholine (DPPC)/cholesterol bilayers and apply it to large-scale simulation studies of lipid membranes. Our CG model is a two-dimensional representation of the membrane, where the individual lipid and sterol molecules are described by pointlike particles. The effective intermolecular interactions used in the model are systematically derived from detailed atomic-scale molecular dynamics simulations using the Inverse Monte Carlo technique, which guarantees that the radial distribution properties of the CG model are consistent with those given by the corresponding atomistic system. We find that the coarse-grained model for the DPPC/cholesterol bilayer is substantially more efficient than atomistic models, providing a speedup of approximately eight orders of magnitude. The results are in favor of formation of cholesterol-rich and cholesterol-poor domains at intermediate cholesterol concentrations, in agreement with the experimental phase diagram of the system. We also explore the limits of the coarse-grained model, and discuss the general validity and applicability of the present approach. PMID- 15527385 TI - Structure formation in films of weakly charged block polyelectrolyte solutions. AB - A mean-field dynamic density functional theory is used to describe a phase diagram of concentrated solutions of weakly charged flexible block polyelectrolytes in a film. Electrostatics is taken into account by applying the local electroneutrality constraint (the Donnan membrane equilibrium approach). In the Donnan limit it is assumed that a salt added to the solution perfectly screens long-range electrostatic interactions. The phase diagram of a solution of a triblock polyelectrolyte in a film as a function of the solvent concentration and the charge of the polyelectrolyte (solvophilic) block is calculated for a given film thickness. The phase behavior of the block polyelectrolyte film arises from the interplay between surface-induced alignment and the electrostatically driven structure formation. The observed mesoscopic structures (lamellar, perforated lamellar, cylindrical, micellar, and mixed phases) are oriented parallel to the surfaces for the considered case of morphologies unfrustrated by the film thickness. Structures with connections between parallel layers (bicontinuous, etc.) are not formed. As a result of surface-induced ordering, the region of ordered phases in a film is wider than in bulk and the phase boundary between ordered and disordered phases is more diffuse. As in the case of unconfined block polyelectrolyte solution, the solution in a film does not follow the lyotropic sequence of phases of such a block copolymer upon increase in the charge of the polyelectrolyte block. Upon changing the charge of the solvophilic copolymer block, transformations of copolymer morphology take place via change in curvature of polymeric domains. Due to confinement of a polyelectrolyte film, no swelling of solvophilic domains is observed. PMID- 15527386 TI - Ab initio study of cubyl chains and networks. AB - The spatial arrangements and physical properties of one- and two-dimensional structures, based on the amazing cubane (C(8)H(8)) molecule, are investigated in detail. In particular, we compute the electronic structure, both by first principle calculations and by semiempirical methods. The elastic and vibrational properties are evaluated as well. All these results are compared with those of the single cubane molecule, in order to elucidate the influence of dimensionality. PMID- 15527387 TI - Sensitized phosphorescence of benzil-doped ladder-type methyl-poly(para phenylene). AB - The delayed luminescence and phosphorescence of ladder-type methyl-poly(para phenylene) (MeLPPP) doped with benzil at a concentration of 20% by weight has been measured. The introduction of benzil leads to a dramatic reduction of the polymer singlet emission. At the same time, a new band with maximum at 611 nm appears, corresponding to the phosphorescence of MeLPPP. The phosphorescence decay on the short time scale is close to an exponential law with a time decay of 15 ms. This indicates that benzil can efficiently sensitize the phosphorescence of the polymer. In addition, a broad and featureless emission is observed in the delayed luminescence spectra of benzil-doped MeLPPP, which is attributed to an exciplex formed between the polymer host and the dopant. We further observe that the delayed fluorescence is enhanced by the addition of benzil. It is concluded that the delayed fluorescence of benzil-doped MeLPPP is mainly due to the annihilation of triplet excitons on the polymer. Finally, efficient triplet triplet energy transfer from the benzil-doped polymer to the red-emitting phosphorescent dye Pt(II)octaethylporphyrin is established. PMID- 15527388 TI - Misfolded free energy surface of a peptide with alphabetabeta motif (1PSV) using the generalized Born solvation model. PMID- 15527389 TI - Ensemble of transition states for two-state protein folding from the eigenvectors of rate matrices. PMID- 15527390 TI - Doppler-free two-photon excitation spectroscopy and the Zeeman effect of the 140 1 band of the S1 1B2u<--S0 1A1g transition of benzene-d6. PMID- 15527392 TI - Comment on "Quasielastic neutron scattering of two-dimensional water in a vermiculite clay" [J. Chem. Phys. 113, 2873 (2000)] and "A neutron spin-echo study of confined water" [J. Chem. Phys. 115, 11299 (2001)]. PMID- 15527391 TI - Geometry relaxation effects for molecules as a result of binding with a positron. PMID- 15527396 TI - Catching up in pharmaceuticals: government policies and the rise of genomics. AB - Genomics--the sequencing of the human genome and the identification of the genetic mechanisms of development and disease--is driving a restructuring of the global pharmaceutical industry. Many policy initiatives in biomedicine are based on the assumption that strength in genomics research will be translated into economic success, that is, that investments in genomics will be a way to catch up in the development of the bio-industries. There is therefore intense competition between nations and regions to establish a strong position in genomics, as shown in the growth of public expenditure on biomedical research in the last decade. This article addresses questions regarding the possibility of smaller countries catching up in the bio-based economy, given its present concentration within a few research-intensive networks and the historical advantage of established knowledge clusters, which exist primarily in the United States. PMID- 15527395 TI - Pharmaceuticals, the state and the global harmonisation process. AB - This article examines how regulatory agencies' mission to protect and promote public health, enshrined in legislation, has been shaped and limited by commitments to the commercial interests of the pharmaceutical industry. It is argued that the regulatory state has become largely a 'competition state' which considers its primary role to be the maintenance of industry's competitive position in world markets. By examining regulatory developments across the EU, Japan and the US, I shall explain how the competition state became a building block for the global harmonisation process. To legitimise the global harmonisation process in terms of their mission to protect and promote public health, regulators claim that it does not lower safety standards and will accelerate the availability of pharmaceutical innovations to patients who need them. However, evidence is presented to suggest that these legitimising claims are not tenable. PMID- 15527397 TI - "Tough but fair"? The active management of the New Zealand drug benefits scheme by an independent Crown agency. AB - For just over a decade, New Zealand has relied on an independent Crown agency to manage the public drug benefits scheme. It was established after a period of industry litigation and unsustainable budgetary increases. The agency has successfully contained prices, saving the equivalent to its originally allocated budget every year, despite a 50% increase in volumes. It shares features with similar agencies elsewhere in the world, particularly in its independence and its operational methodology. Opposition from the industry and ambivalence in the medical community remain matters of concern. The fate of such agencies is inextricably linked to wider regulatory and policy settings in the health sector. PMID- 15527398 TI - India's pharmaceutical industry: hype or high tech take-off? AB - India has built a large pharmaceutical industry through an array of measures in support of domestic firms. The absence of product patents enabled Indian companies to become world leading producers of generic versions of patented drugs. Low costs and a strong engineering tradition continue to sustain competitive strength. The implementation of the World Trade Organization patent regime in 2005 is driving a transformation of the industry. Key elements of the present shake-up include the return of 'big pharma' companies on a large scale and the emergence of several Indian firms that aim to become fully-fledged research-based multinationals. This article provides a description of the development and structure of the Indian pharmaceutical industry and explores questions and challenges arising from its integration into global markets. PMID- 15527399 TI - The subsidy of pharmaceuticals in Australia: processes and challenges. AB - The increasing costs of health care, including new technologies and pharmaceuticals, pose challenges for all countries both in the developed and the developing world. It is essential that the increased expenditure on pharmaceuticals represents value for money and is seen as an investment in health care rather than simply the purchase of the latest released agent. The system in Australia uses a cost-effectiveness approach to guide the decisions as to whether a new drug can be recommended for subsidy. The need for a greater understanding and transparency of the processes is essential in order for a well-informed public debate to occur about the challenges to the system and its sustainability. The relevant issues are discussed in this article to assist that debate. PMID- 15527401 TI - Some drugs more equal than others: pseudo-generics and commercial practice. AB - This article analyses the impact of the Department of Health and Ageing's brand price premium policy for some products listed on the Pharmaceutical Benefits Scheme. The policy, introduced in 1990, allows pharmaceutical companies to charge patients an out-of-pocket expense for post-patent brands of pharmaceuticals. One of the policy's intended goals was to increase consumer awareness of price differentials between competing brands, with a view to encouraging greater use of cheaper generic products. More than fourteen years since its introduction, it is debatable whether the policy has achieved this aim. This article looks at how the brand price premium policy can be exploited by global pharmaceutical giants to entrench big-name brands in the Australian pharmaceutical market and, in some cases, prevent 'true' competition from generic pharmaceuticals. This is being done through the establishment of 'pseudo-generics' that are sourced from the same factory floor as the original product. PMID- 15527402 TI - Patents, pills and politics: the Australia-United States Free Trade Agreement and the Pharmaceutical Benefits Scheme. AB - There is tension between the need of the pharmaceutical innovator for intellectual property protection and the need of society for equitable and affordable access to innovative drugs. The recent Australia-United States Free Trade Agreement provides a nice illustration of this interplay between patents, pills and politics. This article provides a brief history of patent law as applied to pharmaceuticals, describes how the Pharmaceutical Benefits Scheme got caught up in AUSFTA negotiations, analyses the clauses that are likely to impact upon the PBS and describes the political process that reviewed and ultimately amended the AUSFTA. PMID- 15527403 TI - Pharmaceuticals and the consumer movement: the ambivalences of 'patient power'. AB - Consumer and patient advocacy groups (PAGs) are important participants in the politics of pharmaceuticals. Yet very little is known about the precise nature and extent of their influence. It is argued in this article that PAGs fulfil a mixed role within the health system at national and transnational levels, and that they are at times fully incorporated into economic and political power structures. Their frequent dependence on pharma industry funding is of particular concern. PAGs provide a means of direct industry interaction with the final customer, thereby partially bypassing and putting additional pressure on doctors and regulators. The article presents the case for research to establish a better empirical base for discussions about the role of PAGs within contemporary neo liberal governance structures. PMID- 15527404 TI - Community pharmacy in Australia. AB - This article describes the evolution of community pharmacy in the Australian health system, and assesses its current and potential future contribution to health care. A central theme is the unique extent and accessibility of community pharmacy to the public, with a vast and dispersed infrastructure that is funded by private enterprise. The viability of community pharmacy as a retail trade depends on a diversification of its service roles and retention of its product supply roles. Initiatives by the pharmacy profession, the pharmacy industry and the Australian Government are likely to give community pharmacy an increasingly prominent place in health promotion and primary, secondary and tertiary prevention, especially in relation to the management of chronic diseases. PMID- 15527405 TI - Apoptosis in unicellular organisms: mechanisms and evolution. AB - Data about the programmed death (apoptosis) in unicellular organisms, from bacteria to ciliates, are discussed. Firstly apoptosis appeared in lower eukaryotes, but its mechanisms in these organisms are different from the classical apoptosis. During evolution, the apoptotic process has been improving gradually, with reactive oxygen species and Ca2+ playing an essential role in triggering apoptosis. All eukaryotic organisms have apoptosis inhibitors, which might be introduced by viruses. In the course of evolution, caspases and apoptosis-inducing factor appeared before other apoptotic proteins, with so called death receptors being the last among them. The functional analogs of eukaryotic apoptotic proteins take parts in the programmed death of bacteria. PMID- 15527406 TI - Role of platelets and serine proteinases in coupling of blood coagulation and inflammation. AB - In addition to having a key role in thrombogenesis, platelets are actively involved in acute and chronic inflammation: they induce the release of proinflammatory mediators, expose adhesion molecules, and recruit leukocytes. The inflammation-induced expression of tissue factor by endothelium and monocytes leads to production of hemostatic serine proteinases, which can regulate both blood coagulation and the inflammatory response of the body. Serine proteinases activate blood and connective tissue cells and regulate blood coagulation, inflammation, tissue repair, atherogenesis, etc. This review considers new functions of platelets in thrombogenesis and inflammation, stabilization of platelet-platelet and platelet-leukocyte aggregations, receptor functions of tissue factor, proinflammatory properties of hemostatic serine proteinases mediated by proteinase-activated receptors (PAR), activation of transcriptional factors (NFkappaB and other), and antiinflammatory and cytoprotective properties of the anticoagulant proteinase (activated protein C) mediated through binding of the endothelial protein C receptor (EPCR) and cleavage of PAR1. PMID- 15527407 TI - Antibacterial proline-rich oligopeptides and their target proteins. AB - This review presents findings on a new family of antibacterial proline-rich oligopeptides--pyrrhocoricin, drosocin, apidaecin, and formaecin--isolated from insects. The functional and physicochemical properties of proline-rich oligopeptides are considered, a role of proline in their antibacterial activity is discussed, and experimental evidence is given in favor of the ability of these oligopeptides to suppress metabolism of bacteria by means of stereospecific interaction with heat shock protein DnaK and inhibition of DnaK-dependent protein folding. Binding of the peptides under investigation with DnaK correlates with their antibacterial activity. Evidence that pyrrhocoricin, drosocin, apidaecin, and formaecin are nontoxic for human and animal cells serves as a prerequisite for their use as novel antibiotic drugs. PMID- 15527408 TI - Subtilisin protein inhibitor from potato tubers. AB - A protein with molecular weight of 21 kD denoted as PKSI has been isolated from potato tubers (Solanum tuberosum L., cv. Istrinskii). The isolation procedure includes precipitation with (NH4)2SO4, gel chromatography on Sephadex G-75, and ion-exchange chromatography on CM-Sepharose CL-6B. The protein effectively inhibits the activity of subtilisin Carlsberg (Ki = 1.67 +/- 0.2 nM) by stoichiometric complexing with the enzyme at the molar ratio of 1 : 1. The inhibitor has no effect on trypsin, chymotrypsin, and the cysteine proteinase papain. The N-terminal sequence of the protein consists of 19 amino acid residues and is highly homologous to sequences of the known inhibitors from group C of the subfamily of potato Kunitz-type proteinase inhibitors (PKPIs-C). By cloning PCR products from the genomic DNA of potato, a gene denoted as PKPI-C2 was isolated and sequenced. The N-terminal sequence (residues from 15 to 33) of the protein encoded by the PKPI-C2 gene is identical to the N-terminal sequence (residues from 1 to 19) of the isolated protein PKSI. Thus, the inhibitor PKSI is very likely encoded by this gene. PMID- 15527409 TI - Immunochemical approach to the problem of differential determination of natural forms of abscisic acid. AB - An original modification of the standard ELISA procedure for differential determination of different forms of abscisic acid (ABA) is proposed. It is shown that endogenous forms of ABA may be quantitatively determined in plant tissues subjected to minimal treatment, without purification of the hormones and their chemical modification. The modification has been approved when analyzing changes in the content of different ABA forms in plant tissues differing in physiological activity. Quantitative differential determination of changes in the content of different ABA forms has been performed in ovaries of Triticum aestivum L. and Taraxacum officinale Web. in the period of activity of the ovule (from the moment of its activation to the beginning of division). It is shown that, despite the different types of reproduction in the species studied (amphimixis and apomixis), the time course of changes in the content of different forms of ABA in ovaries is similar, which is suggestive of a correlation between the activity of endogenous hormonal system and chronology of main events (e.g., the beginning of endospermogenesis) of the reproductive cycle. PMID- 15527410 TI - Dynamics of DNA-protein complex formation in rat liver during induction by phenobarbital and triphenyldioxane. AB - CYP2B gene expression in liver of rats treated with phenobarbital and triphenyldioxane at early stage of induction (40 min-18 h) was studied using electrophoretic mobility shift assay (EMSA) and RT-PCR. During first 6 h after induction, differences in the dynamics of formation of DNA-protein complexes were shown for each inducer. Later (18 h after induction), the intensity pattern of these complexes became the same for both phenobarbital and triphenyldioxane treated animals. This suggests the existence of specific signaling for each inducer only in early stages of CYP2B activation. Increase in nuclear protein (possible transcription factor) binding to Barbie-box regulatory sequence of CYP2B genes was accompanied by their increased expression. Thus, we have demonstrated for the first time that early stages of induction (40 min and 3 h after administration of phenobarbital and triphenyldioxane, respectively) are accompanied by activation of nuclear proteins that can bind to Barbie-box element of CYP2B. Although various chemical inducers cause distinct activation of such binding, this process involves activation of gene transcription. PMID- 15527411 TI - Influence of obstructive cholestasis and sex hormones on the ratio of mRNA of two alternative prolactin receptor isoforms in rat hepatocytes. AB - The effects of obstructive cholestasis and sex hormones on the total content of prolactin receptor mRNA and ratio of mRNAs of its short and long isoforms have been studied in rat hepatocytes. Obstructive cholestasis caused insignificant changes in total content of prolactin receptor mRNA, but the proportion of mRNA of the long isoform increased. Comparison of prolactin receptor mRNA levels in gonadectomized and intact animals revealed opposite effects of male and female sex hormones on total mRNA, but both groups of these hormones increased the proportion of prolactin receptor short form mRNA. Changes in ratio of mRNA of receptor isoforms found in rat hepatocytes under obstructive cholestasis did not depend on levels of sex hormones. Obstructive cholestasis and sex hormones are suggested to regulate the content of long and short prolactin receptor isoforms in hepatocytes independently. PMID- 15527412 TI - Plasmid pRARE as a vector for cloning to construct a superproducer of the site specific nickase N.BspD6I. AB - The gene of methylase M.SccL1I that protects DNA against hydrolysis with the nickase N.BspD6I was inserted into plasmid pRARE carrying genes of tRNA, which are rare in E. coli. The insertion of the gene sscML1I into pRARE was reasoned by incompatibility of pRARE and the plasmid carrying the gene sscML1I, because both plasmids contained the same ori-site. Upon transformation of E. coli TOP10F cells with both the recombinant plasmid pRARE/MSsc and the expression vector pET28b containing the nickase gene bspD6IN under the phage T7 promoter, a strain of E. coli was obtained which produced 7 x 10(5) units of the nickase N.BspD6I per 1 g wet biomass, and this yield was two orders of magnitude higher than the yield of the enzyme from the strain free of pRARE/MSsc. PMID- 15527413 TI - H1 histone in developing and aging coleoptiles of etiolated wheat seedlings. AB - It has been established that the DNA and H1 histone contents in aged coleoptile of 8-day-old etiolated wheat seedling are about 40 and 30%, respectively, lower than those in young seedlings. H1 histone in wheat seedlings is represented as six electrophoretically different subfractions. The ratios of H1 histone subfractions in wheat coleoptile and initial leaf are similar. In contrast to some animal cells, apoptosis in wheat coleoptile is not accompanied by changes in the set and ratios of H1 histone subfractions. Aging of coleoptiles is associated with a progressive diminution of the H1 histone and DNA contents. H1 histone/DNA ratio in aged coleoptile is 1.5-2-fold higher than that in the young organs. Therefore, the content of H1 histone in chromatin of coleoptile decreases with age more slowly than DNA content. PMID- 15527414 TI - Interaction of hydrogen peroxide with ribulose-1,5-bisphosphate carboxylase/oxygenase from rice. AB - The properties of rice-derived ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) in different concentrations of hydrogen peroxide (H2O2) solutions have been studied. The results indicate that at low H2O2 concentrations (0.2-10 mM), the properties of rubisco (e.g., carboxylase activities, structure, and susceptibility to heat denaturation) change slightly. However, at higher H2O2 concentrations (10-200 mM), rubisco undergoes an unfolding process, including the loss of secondary and tertiary structure, forming extended hydrophobic interface, and leading to cross-links between large subunits. High concentrations of H2O2 can also result in an increase in susceptibility of rubisco to heat denaturation. Further pre-treatments with or without reductive reagents to rubisco show that the disulfide bonds in rubisco help to protect the enzyme from damage by H2O2 as well as other reactive oxygen species. PMID- 15527415 TI - Influence of seizures on lipids of homogenate and neuronal and glial nuclei of rat neocortex. AB - Lipid composition of homogenate and neuronal and glial nuclei of the brain cortex of Wistar rats was studied under normal conditions and after seizures induced by injection of picrotoxin. Seizures increased contents of lysophosphatidylcholine, sphingomyelin, and total phospholipids in the homogenate. In neuronal nuclei contents of total phospholipids, sphingomyelin, phosphatidylcholine, and phosphatidylserine decreased, and contents of free fatty acids and lysophosphatidylcholine increased. In glial nuclei content of total phospholipids decreased and content of free fatty acids increased. The role of changes in the lipid composition of the neocortex cells during seizures and the involvement of lipid messengers in signal mechanisms are discussed. PMID- 15527416 TI - Comparative study of structure and activity of cytotoxins from venom of the cobras Naja oxiana, Naja kaouthia, and Naja haje. AB - Cytotoxins are positively charged polypeptides that constitute about 60% of all proteins in cobra venom; they have a wide spectrum of biological activities. By CD spectroscopy, cytotoxins CT1 and CT2 Naja oxiana, CT3 Naja kaouthia, and CT1 and CT2 Naja haje were shown to have similar secondary structure in an aqueous environment, with dominating beta-sheet structure, and to vary in the twisting angle of the beta-sheet and the conformation of disulfide groups. Using dodecylphosphocholine micelles and liposomes, CT1 and CT2 Naja oxiana were shown to incorporate into lipid structures without changes in the secondary structure of the peptides. The binding of CT1 and CT2 Naja oxiana with liposomes was associated with an increase in the beta-sheet twisting and a sign change of the dihedral angle of one disulfide group. The cytotoxins were considerably different in cytotoxicity and cooperativity of the effect on human promyelocytic leukemia cells HL60, mouse myelomonocytic cells WEHI-3, and human erythroleukemic cells K562. The most toxic CT2 Naja oxiana and CT3 Naja kaouthia possessed low cooperativity of interaction (Hill coefficient h = 0.6-0.8), unlike 10-20-fold less toxic CT1 and CT2 Naja haje (h = 1.2-1.7). CT1 Naja oxiana has an intermediate position on the cytotoxicity scale and is characterized by h = 0.5 0.8. The cytotoxins under study induced necrosis of HL60 cells and failed to activate apoptosis. The differences in cytotoxicity are supposed to be related not with features of the secondary structure of the peptides, but with interactions of side chains of variable amino acid residues with lipids and/or membrane proteins. PMID- 15527417 TI - Expression and characterization of hepatitis B surface antigen in transgenic potato plants. AB - Transgenic potato plants expressing the gene of hepatitis B surface antigen (HBsAg) under the control of the double promoter of 35S RNA of cauliflower mosaic virus (CaMV 35SS) and the promoter of patatin gene of potato tubers have been obtained. Biochemical analysis of the plants was performed. The amount of HBsAg in leaves, microtubers, and tubers of transgenic potatoes growing in vitro and in vivo was 0.005-0.035% of the total soluble protein. HBsAg content reached 1 microg/g in potato tubers and was maximal in plants expressing the HBsAg gene under the control of CaMV 35SS promoter. In transgenic plants expressing HBsAg gene under the control of tuber-specific patatin promoter, HBsAg was found only in microtubers and tubers and was absent in leaves. Western blot analysis of HBsAg eluted from immunoaffinity protein A-Sepharose matrix has been performed. The molecular weight of HBsAg peptide was approximately 24 kD, which is in agreement with the size of the major protein of the envelope of hepatitis B virus. Using gel filtration, it was determined that the product of HBsAg gene expression in potato plants is converted into high-molecular-weight multimeric particles. Therefore, as well as in recombinant HBsAg-yeast cells, assembling of HBsAg monomers into immunogenic aggregates takes place in HBsAg-transgenic potato, which can be used as a source of recombinant vaccine against hepatitis B virus. PMID- 15527418 TI - Uncoupling effect of fatty acids in halo- and alkalotolerant bacterium Bacillus pseudofirmus FTU. AB - Natural uncouplers of oxidative phosphorylation, long-chain non-esterified fatty acids, cause uncoupling in the alkalo- and halotolerant bacterium Bacillus pseudofirmus FTU. The uncoupling effect in the bacterial cells was manifested as decrease of membrane potential and increase of respiratory activity. The membrane potential decrease was detected only in bacterial cells exhausted by their endogenous substrates. In proteoliposomes containing reconstituted bacterial cytochrome c oxidase, fatty acids caused a "mild" uncoupling effect by reducing membrane potential only at low rate of membrane potential generation. "Free respiration" induced by the "mild" uncouplers, the fatty acids, can be considered as possible mechanism responsible for adaptation of the bacteria to a constantly changed environment. PMID- 15527419 TI - Use of plant-derived protein hydrolysates for enhancing growth of Bombyx mori (silkworm) insect cells in suspension culture. AB - The successful suspension culture of the Bombyx mori (silkworm) cell lines Bm5 and BmN4 without FBS (fetal-bovine serum) was first realized in Sf-900 II SFM (serum-free medium) (Gibco BRL, Rockville, MD, U.S.A.) supplemented with a plant derived protein hydrolysate. The addition of 0.5% HyPep 1510 (Difco Co., Detroit, MI, U.S.A.) and 10 mM glutamine to Sf-900 II SFM at 4 days of culture was found effective in increasing the cell concentration to 8.5x10(6) cells/ml. The replacement of medium with Sf-900 II SFM supplemented with 0.5% HyPep 1510 at 6 days of culture increased the cell concentration by 1.1x10(7) cells/ml. When Sf 900 II SFM was supplemented with 0.5% Hypep 1510, 16 days, which was half of the conventional adaptation time, was sufficient for the B. mori cell line to adapt to SFM and shear stress while maintaining a stable viability. The beta galactosidase activity in Sf-900 II SFM supplemented with 0.5% Hypep 1510 was 4.9x10(3) units/ml, which was 2-fold higher than that of the FBS-supplemented medium. By SDS/PAGE, only the band corresponding to beta-galactosidase was detected in the sample from the media supplemented with plant-derived protein hydrolysates, while thick bands corresponding to proteins having lower molecular masses than beta-galactosidase were detected in samples from the FBS-supplemented media. These results suggest that plant-derived protein hydrolysates are promising FBS substitutes for enhancing the growth of B. mori cells and facilitating the purification of recombinant proteins produced by baculovirus infection. PMID- 15527420 TI - A novel human complement-related protein, C1r-like protease (C1r-LP), specifically cleaves pro-C1s. AB - The availability of the human genome sequence allowed us to identify a human complement-related, C1r-like protease gene (c1r-LP) located 2 kb centromeric of the C1r gene (c1r). Compared with c1r, c1r-LP carries a large deletion corresponding to exons 4-8 of c1r. The open reading frame of the C1r-LP cDNA predicts a 50 kDa modular protein displaying 52% amino acid residue identity with the corresponding regions of C1r and 75% identity with a previously described murine C1r-LP. The serine protease domain of C1r-LP, despite an overall similarity with the AGY group of complement serine proteases, has certain structural features characteristic of C2 and factor B, thus raising interesting evolutionary questions. Northern blotting demonstrated the expression of C1r-LP mRNA mainly in the liver and ELISA demonstrated the presence of the protein in human serum at a concentration of 5.5+/-0.9 microg/ml. Immunoprecipitation experiments failed to demonstrate an association of C1r-LP with the C1 complex in serum. Recombinant C1r-LP exhibits esterolytic activity against peptide thioesters with arginine at the P1 position, but its catalytic efficiency (kcat/K(m)) is lower than that of C1r and C1s. The enzymic activity of C1r-LP is inhibited by di-isopropyl fluorophosphate and also by C1 inhibitor, which forms stable complexes with the protease. Most importantly, C1r-LP also expresses proteolytic activity, cleaving pro-C1s into two fragments of sizes identical with those of the two chains of active C1s. Thus C1r-LP may provide a novel means for the formation of the classical pathway C3/C5 convertase. PMID- 15527421 TI - Functional dichotomy of A20 in apoptotic and necrotic cell death. AB - ROS (reactive oxygen species) play important roles in the progression of a number of human pathologies. ROS promote cell death, but can also induce gene transcription. The transcription factor NF-kappaB (nuclear factor kappaB) plays a critical role in oxidative stress responses. One of the proteins regulated by NF kappaB is the zinc-finger protein A20. In TNF (tumour necrosis factor)-alpha signalling, NF-kappaB induction of A20 leads to increased cell survival. In the present paper, we show that in response to oxidative stress, A20 actually enhances cell death by necrosis, but not by apoptosis. Exposure of cells to ROS leads to the up-regulation of A20 which acts via a negative-feedback loop to block NF-kappaB activation and cellular survival. Silencing of A20 by RNAi (RNA interference) increases both the induction of NF-kappaB and the subsequent survival of cells exposed to high doses of oxidative stress, which, in untreated cells, promotes death by necrosis. Cells which express high basal levels of A20 are less protected from oxidative-stress-induced cell death when compared with cells with lower A20 expression. We also show that A20 regulates NF-kappaB by blocking the degradation of IkappaB (inhibitory protein kappaB) alpha. These data highlight a novel role for A20 in oxidative stress responses by terminating NF kappaB-dependent survival signalling and thus sensitizing cells to death by necrosis. PMID- 15527422 TI - Quantitative determination of the binding of beta2-glycoprotein I and prothrombin to phosphatidylserine-exposing blood platelets. AB - The plasma protein beta2GPI (beta2-glycoprotein I) has been proposed to mediate phagocytosis of apoptotic cells and to play a role in the antiphospholipid syndrome. This suggestion is based mainly on the presumption that beta2GPI has an appreciable interaction with PS (phosphatidylserine)-exposing cell membranes. However, quantitative data on the binding of beta2GPI to PS-exposing cells under physiologically relevant conditions are scarce and conflicting. Therefore we evaluated the binding of beta2GPI to PS-expressing blood platelets. Flow cytometry showed that binding of beta2GPI is negligible at physiological ionic strength, in contrast with significant binding occurring at low ionic strength. Binding parameters of beta2GPI and (for comparison) prothrombin were quantified by ellipsometric measurement of protein depletion from the supernatant following incubation with platelets. At low ionic strength (20 mM NaCl, no CaCl2), a dissociation constant (K(d)) of 0.2 microM was found for beta2GPI, with 7.4x10(5) binding sites per platelet. Under physiologically relevant conditions (120 mM NaCl and 3 mM CaCl2), binding of beta2GPI was not detectable (extrapolated K(d)>80 microM). Prothrombin binding (at 3 mM CaCl2) was much less affected by ionic strength: K(d) values of 0.5 and 1.4 muM were observed at 20 and 120 mM NaCl respectively. The low affinity and the presence of many lipid-binding proteins in plasma that can compete with the binding of beta2GPI suggest that only a small fraction (<5%) of the binding sites on PS-exposing blood cells are likely to be occupied by beta2GPI. These findings are discussed in relation to the alleged (patho-)physiological functions of beta2GPI. PMID- 15527423 TI - Gelsolin binds to polyphosphoinositide-free lipid vesicles and simultaneously to actin microfilaments. AB - Gelsolin is a calcium-, pH- and lipid-dependent actin filament severing/capping protein whose main function is to regulate the assembly state of the actin cytoskeleton. Gelsolin is associated with membranes in cells, and it is generally assumed that this interaction is mediated by PPIs (polyphosphoinositides), since an interaction with these lipids has been characterized in vitro. We demonstrate that non-PPI lipids also bind gelsolin, especially at low pH. The data suggest further that gelsolin becomes partially buried in the lipid bilayer under mildly acidic conditions, in a manner that is not dependent of the presence of PPIs. Our data also suggest that lipid binding involves a number of sites that are spread throughout the gelsolin molecule. Linker regions between gelsolin domains have been implicated by other work, notably the linker between G1 and G2 (gelsolin domains 1 and 2 respectively), and we postulate that the linker region between the N-terminal and C-terminal halves of gelsolin (between G3 and G4) is also involved in the interaction with lipids. This region is compatible with other studies in which additional binding sites have been located within G4-6. The lipid-gelsolin interactions reported in the present paper are not calcium dependent, and are likely to involve significant conformational changes to the gelsolin molecule, as the chymotryptic digest pattern is altered by the presence of lipids under our conditions. We also report that vesicle-bound gelsolin is capable of binding to actin filaments, presumably through barbed end capping. Gelsolin bound to vesicles can nucleate actin assembly, but is less active in severing microfilaments. PMID- 15527424 TI - The central part of the 5.8 S rRNA is differently arranged in programmed and free human ribosomes. AB - A sequence-specific modification of the human 5.8 S rRNA in isolated 60 S subunits, non-programmed 80 S ribosomes and ribosomes complexed with mRNA and tRNAs was studied with the use of a derivative of the nonaribonucleotide UCUGUGUUU bearing a perfluorophenylazide group on the C-5 atom of the 5'-terminal uridine. Part of the oligonucleotide moiety of the derivative was complementary to the 5.8 S rRNA sequence ACACA in positions 82-86 flanked by two guanines at the 5'-terminus. The target for the cross-linking was identified as nucleotide G89 on the 5.8 S RNA. In addition, several ribosomal proteins were modified by the oligonucleotide derivative bound to the 5.8 S rRNA and proteins L6 and L8 were among them. Application of these results to known cryo-electron microscopy images of eukaryotic 60 S subunits made it possible to suggest that the central part of the 5.8 S rRNA containing the sequence 82-86 and proteins L6 and L8 are located at the base of the L1 stalk of the 60 S subunit. The efficacy of cross linking in non-programmed 80 S ribosomes was much lower than in isolated 60 S subunits and in programmed 80 S ribosomes. We suggest that the difference in the accessibilities of the central part of the 5.8 S rRNA in the programmed and non programmed 80 S ribosomes is caused by a conformational switch that seems to be required to dissociate the 80 S ribosomes into the subunits after termination of translation to allow initiation of translation of a new template. PMID- 15527426 TI - Adjunctive bright light in non-seasonal major depression. AB - OBJECTIVE: Bright light treatment is an established treatment for Seasonal Affective Disorder, but in non-seasonal depression research results have been contrasting. METHOD: This study was designed as a 5-week controlled, double blind, parallel trial in out-patients with a diagnosis (DSM-IV) of non-seasonal major depression, randomized to either active treatment (white light, 10 000 lux, 1 h daily) or placebo treatment (red light, 50 lux, 30 min daily) and concomitant treatment with sertraline in both groups. RESULTS: One hundred and two patients were included in the study. Analyses showed that on all used scales the reduction in depression scores was larger in the bright light group than in the dim light group, and this reached statistical significance on all observer rating scales and on the SCL-90R self-assessment scale. The HAM-D6 was the most sensitive scale to measure improvement at endpoint. CONCLUSION: The study results support the use of bright light as an adjunct treatment to antidepressants in non-seasonal depression. PMID- 15527428 TI - Malassezia yeasts in the pathogenesis of atopic dermatitis. AB - Atopic dermatitis is a common skin condition, the aetiology of which is multifactorial, involving genetic, immunological and environmental factors. In recent years, it has been suggested that various microbial organisms may also be involved in the pathogenesis of the disease. Yeasts belonging to the Malassezia genus have received particular attention. These yeasts, known to be a part of the normal skin flora, have been shown to be capable of inducing immunoglobulin (Ig)E mediated and T-cell mediated immune responses postulated to contribute to chronic inflammation in the skin, particularly in the head and neck region, where colonization with Malassezia is the greatest. Considerable evidence now exists to support this idea, raising the possibility that specific antifungal therapy may be a useful treatment measure in some atopic patients who have a head and neck pattern of dermatitis. PMID- 15527429 TI - Iontophoresis with glycopyrrolate for the treatment of palmoplantar hyperhidrosis. AB - To determine the comparative efficacy of tap water iontophoresis to iontophoresis with the anticholinergic glycopyrrolate, we undertook a single-blinded right-left comparison study in 20 patients with palmoplantar hyperhidrosis. Most patients had their palms treated and one patient had the soles treated. We compared the duration of symptom relief following iontophoresis with glycopyrrolate unilaterally to iontophoresis with glycopyrrolate bilaterally. Patients filled in daily efficacy assessment cards. Each palm was rated as 'dry', 'slightly wet', 'moderately wet' or 'very wet'. Following treatment with unilateral tap water iontophoresis, unilateral glycopyrrolate and bilateral glycopyrrolate, patients reported hand dryness for a median of 3, 5 and 11 days, respectively. As the data was paired, treatment differences were analysed using a sign-rank test. Bilateral glycopyrrolate was superior to both unilateral glycopyrrolate and tap water in most patients. Unilateral glycopyrrolate was superior to tap water in most patients. All differences between groups were found to be statistically significant. We postulate that the increased efficacy of bilateral glycopyrrolate when compared with unilateral glycopyrrolate relates to its systemic absorption. We conclude that glycopyrrolate iontophoresis is more effective than tap water iontophoresis in the treatment of palmoplantar hyperhidrosis and that glycopyrrolate iontophoresis has both local and systemic effects on perspiration. PMID- 15527430 TI - Case of recalcitrant necrobiotic xanthogranuloma. AB - A 53-year-old woman with necrobiotic xanthogranuloma presented as infiltrated eyelid plaques, with later development of arm and thigh lesions. Clinical features, including association with immunoglobulin G-kappa paraprotein, and pathological findings were typical of this disorder. Treatment for 15 months with varying combinations of prednisone and multiple chemotherapeutic agents (melphalan, cyclophosphamide and chlorambucil) has led to minimal or no improvement in the clinical lesions or paraprotein. The case demonstrates some of the difficulties in managing this unusual disorder. PMID- 15527431 TI - Gallbladder vasculitis associated with cutaneous leucocytoclastic vasculitis. AB - A 72-year-old woman presented with a 1-week history of a painful, purpuric, pruritic rash on her legs, buttocks and arms. Skin biopsy revealed histological features typical of leucocytoclastic vasculitis. She was admitted, her usual medications were withheld, and she was commenced on ibuprofen and loratadine. The patient had undergone a laparoscopic cholecystectomy 2 months prior to her rash appearing. She had been having upper abdominal pain for 2 years and, following a more severe acute episode, an abdominal ultrasound scan had revealed a solitary cholelithiasis. Histology of the gallbladder revealed acute fibrinoid vasculitis in two small arteries, on a background of chronic cholecystitis. A mild postoperative wound infection was treated with a short course of cephalexin and no other investigations were conducted. As an inpatient, the patient's rash improved, but she progressed to develop systemic vasculitis and acute renal failure. Renal biopsy showed focal necrotizing glomerulonephritis, consistent with vasculitis. She was subsequently commenced on cyclophosphamide 100 mg daily, prednisolone 50 mg daily and one prophylactic trimethoprim/sulphamethoxazole (160 mg/800 mg) tablet 3 days/week. Following discharge, the patient's cutaneous vasculitis eventually resolved and renal function gradually improved. Her prednisolone was gradually reduced to 7 mg daily and cyclophosphamide was weaned, then substituted with azathioprine 100 mg daily. PMID- 15527432 TI - Case of the Brooke-Spiegler syndrome. AB - A 36-year-old woman presented with lesions on her scalp, face and trunk. Histopathological examination of these lesions demonstrated facial trichoepithelioma, and scalp cylindroma. A solitary nodule on the trunk had features of cylindroma, spiradenoma and trichoepithelioma, a previously unreported occurrence. Based on the clinical picture, the diagnosis of Brooke Spiegler syndrome was established. Genetic studies confirmed the diagnosis, demonstrating a splice site mutation, designated 1518+2T>C, on the CYLD1 gene of chromosome 16q12-q13. PMID- 15527433 TI - Use of gloves in protection from diallyl disulphide allergy. AB - Contact dermatitis, particularly affecting the fingertips, is a recognized presentation of garlic allergy. There have been no recommendations in the literature with respect to the type of gloves that offer the best protection against diallyl disulphide, the major allergen in garlic and onion. In fact, we have found that diallyl disulphide penetrates most commercially available glove types. Silver laminate gloves offered only slightly better protection. PMID- 15527434 TI - Homozygous familial hypercholesterolaemia presenting with cutaneous xanthomas: response to liver transplantation. AB - A 19-month-old girl with cutaneous xanthomas was diagnosed with homozygous familial hypercholesterolaemia, treated with atorvastatin, plasma exchange and finally at the age of 3.5 years, with liver transplantation. Her serum cholesterol levels fell to normal and the xanthomas regressed following liver transplantation. She remains well 17 months post transplant. PMID- 15527435 TI - Textile dye allergic contact dermatitis following paraphenylenediamine sensitization from a temporary tattoo. AB - Textile dye allergy is frequently caused by azo dyes, which can cross-react with structurally similar compounds, including paraphenylenediamine. A case of allergic contact dermatitis to azo textile dyes, presenting principally as a sock dermatitis, is presented. The patient also gave a history of an episode of scalp dermatitis consistent with contact allergy to paraphenylenediamine in hair dye. It is proposed that paraphenylenediamine sensitization from a temporary skin tattoo may have been the primary sensitizing event for these reactions. PMID- 15527436 TI - Bowen's disease of the nail bed. AB - A 77-year-old man presented with a 2-year history of left thumbnail dystrophy, purulent discharge and aching pain. There was no history of trauma. On examination there was a linear area of nail dystrophy and dyschromia of the nail bed. The nail plate was thinned proximally. No other digits were affected. There was no lymphadenopathy. X-ray of the digit was normal. Longitudinal nail biopsy revealed Bowen's disease along the length of the nail bed and nail matrix specimen. After avulsion of the nail plate, the nail matrix and tumour were fully excised. The defect was repaired with a split-skin graft. His symptoms were relieved. PMID- 15527438 TI - Simplified dermoscopic diagnosis of acral melanocytic lesions: mountains and valleys. PMID- 15527437 TI - Allergic contact dermatitis to a sanitary pad. AB - A 13-year-old girl presented with a recurring vulval dermatitis that started several days after first using a sanitary pad. The rash settled but recurred with subsequent use of the same sanitary pads. Patch testing revealed a marked sensitivity (3+) to colophony (rosin). She denied previous reactions to adhesive tape/sticking plaster. During patch testing she also developed reactions to a black permanent waterproof marking pen used to mark the site of patch tests. The ink of this pen also contained colophony. PMID- 15527439 TI - Localized vulvar syringoma causing vulval pruritus and venerophobia. PMID- 15527440 TI - Axillary perifollicular xanthomatosis resembling Fox-Fordyce disease. PMID- 15527445 TI - Stroke recurrence within the first year following cerebral infarction--Tottori University Lacunar Infarction Prognosis Study (TULIPS). AB - OBJECTIVES: To identify determinants of recurrence after ischemic stroke in the Japanese population. MATERIALS AND METHODS: We enrolled 885 patients with acute ischemic stroke that had been admitted to our community hospitals. A total of 831 cases were followed for 1 year after the index stroke. Patients were assigned to one of the ischemic stroke subtype groups based on the NINDS Stroke Data Bank criteria. RESULTS: Rates of stroke recurrence were significantly different among stroke subtype groups: 14.4% in cardioembolic infarction, 7.3% in atherothrombotic infarction (ATI), 6.2% in lacunar infarction (LI) and 7.8% in infarction of uncertain cause. Previous history of stroke was the predictor of stroke recurrence for the groups of ATI and LI patients, and diabetes mellitus was the predictor of recurrence for the group of LI patients. CONCLUSION: The rate of recurrence and risk factors for stroke recurrence are different by stroke subtypes. PMID- 15527446 TI - Association of lacunar infarcts with small artery and large artery disease: a comparative study. AB - OBJECTIVES: Patients with lacunar infarcts (LI) and ipsilateral large artery disease (LAD) greater than 50% must be classified according to the Trial of ORG 10172 in Acute Stroke Treatment (TOAST) criteria as strokes of undetermined etiology. The purpose of this study was to compare the vascular risk factors, clinical symptoms, and outcome characteristics of LI associated with LAD with those patients with LI who fulfilled the TOAST criteria of small artery disease (SAD). METHODS: Among 1754 consecutive first ever stroke patients admitted to our department, we analyzed age, gender, vascular risk factors (hypertension, diabetes, ischemic heart disease, arterial peripheral disease, hypercholesterolemia, smoking, alcohol, or illicit drug use), clinical data (motor or sensitive deficit and presence of dysarthria), and outcome (hospitalization length, in-hospital medical complications rate, need of rehabilitation, treatment at discharge, in-hospital mortality, and modified Rankin Scale at discharge) of those patients classified as LI associated with LAD as compared with those with SAD. RESULTS: After a strict application of the TOAST criteria, we found 144 patients with LI associated with SAD and 73 patients with LI associated with LAD. Univariate analysis showed statistical differences in gender (OR: 0.46; 95% CI: 0.23-0.89; P = 0.014), past history of ischemic heart disease (OR: 0.32; 95% CI: 0.13-0.78; P = 0.004), and smoking (OR: 0.56; 95% CI: 0.31-1.04; P = 0.048). After logistic regression analysis only ischemic heart disease (OR: 0.31; 95% CI: 0.11-0.78; P = 0.013), and gender (OR: 0.51; 95% CI: 0.28-0.98; P = 0.05) showed statistical differences. During the follow-up, six patients (all with LI associated with LAD) experienced stroke recurrences (OR: 0.32; 95% CI: 0.26-0.39; P < 0.001). CONCLUSIONS: 1) There are no differences in clinical presentation and in-hospital outcome between patients with LI associated with SAD and patients with LI associated with LAD. 2) Risk factors are very similar in both groups, and the only differences observed (gender and ischemic heart disease) are related to the atherosclerotic factor. 3) Stroke recurrence seems to be more frequent in LI associated with LAD than in LI associated with SAD, but large follow-up studies are needed to be able to decide whether clinical recurrence of stroke allows to differentiate both clinical entities. PMID- 15527447 TI - The FABP2 gene polymorphism in cerebrovascular disease. AB - OBJECTIVE: Earlier studies have shown that the fatty acid binding protein 2 (FABP2) T54 allele is associated with dyslipidemia, which in turn correlates with the incidence of cerebrovascular disease (CVD). To assess whether the FABP2 gene A54T polymorphism is associated with an increased risk of CVD we undertook a case control study. MATERIALS AND METHODS: A total of 407 patients diagnosed with acute CVD and 158 control subjects were genotyped for the A54T polymorphism using a PCR-RFLP method. RESULTS: Allele and genotype frequencies of the FABP2 A54T polymorphism did not differ between subjects with acute CVD (TT, 9.6%; TA, 41.0%; AA, 49.4%) and controls (TT, 7.6%; TA, 41.1%; AA, 51.3%; P = ns) or in the following subgroups of CVD compared with controls: non-cardioembolic infarction (n = 252), intracerebral hemorrhage (n = 23), and cardioembolic infarction (n = 91). In transient ischemic attacks (TIAs) (n = 41) the combined TT and TA genotype frequency (TT + TA, 65.9%) was more frequent than in controls (48.7%) (P = 0.05). Furthermore, the TT genotype was more frequent in non-smoking patients under the age of 70 (n = 77) with a non-cardioembolic infarction (TT, 18.2%) compared with controls (7.6%) (P = 0.024). CONCLUSIONS: Our findings suggest an involvement of the FABP2 (A54T) gene polymorphism in the pathogenesis of CVD. The FABP2 T54 allele appears to be a genetic susceptibility marker for TIA and non cardioembolic infarction at younger onset. PMID- 15527448 TI - Diffusion-weighted MRI during early global cerebral hypoxia: a predictor for clinical outcome? AB - OBJECTIVES: As prognostic assessment of prolonged cerebral hypoxia is often difficult on clinical grounds, a tool for an early prognosis of clinical outcome is desirable. PATIENTS AND METHODS: In a prospective study, we investigated the prognostic value of diffusion-weighted MRI (DWI) in 12 patients within 36 h after global cerebral hypoxia. Results of DWI including apparent diffusion coefficient maps (ADC) were analyzed and related to the clinical outcome after 6 months, in comparison with conventional magnetic resonance imaging (cMRI). RESULTS: Three patients with a short resuscitation time showed normal findings in cMRI and DWI and a good recovery. In seven patients, DWI revealed multiple large hyperintense areas although cMRI was normal. In two patients, large diffuse lesions were observed in DWI which were also found in cMRI. All of these nine patients developed a vegetative state in the follow-up examination. CONCLUSION: Pathological DWI during the early phase after cerebral hypoxia might be superior to cMRI as a predictor of a worse clinical outcome. PMID- 15527449 TI - Spectrum of cutaneous hyperalgesias/allodynias in neuropathic pain patients. AB - OBJECTIVES: The aim of this study was to discern the pathophysio-logical bases for neuropathic hyperalgesias. METHODS: In this study, neurological and neurophysiological evaluation of 132 consecutive hyperalgesia patients using rigorous clinical and laboratory protocols were carried out. RESULTS: Two discrete semeiologic entities emerged: classic neurological vs atypical, fulfilling taxonomically complex regional pain syndrome (CRPS) II and I, respectively. The classic group (34.9%) exhibited sensorimotor patterns restricted to nerve distribution and documented nerve fiber dysfunction. Among them four (3.03%) had sensitization of C-nociceptors, seven (5.3%) had central release of nociceptive input, and 35 (26.52%) probable ectopic nerve impulse generation. The atypical group (65.1%) displayed weakness with interrupted effort; non-anatomical hypoesthesia and hyperalgesia; hypoesthesia or paresis reversed by placebo, or atypical abnormal movements, and physiological normality of motor and sensory pathways. CONCLUSIONS: Spatiotemporal features of neuropathic hyperalgesia constitute key criteria for differential diagnosis between CRPS II and I and, together with other behavioral sensorimotor features, signal psychogenic pseudoneurological dysfunction vs structural neuropathology. 'Neuropathic' hyperalgesias may reflect neuropathological or psychopathological disorders. PMID- 15527450 TI - CD45RA+ ICAM-3+ lymphocytes in interferon-beta1b-treated and -untreated patients with relapsing-remitting multiple sclerosis. AB - OBJECTIVES: Multiple sclerosis (MS) is believed to be an autoimmune disease of the human central nervous system mediated by autoreactive T cells. Interferon beta1b (IFN-beta1b) has been shown to be effective in reducing disease activity defined by clinical and magnetic resonance imaging (MRI) criteria in relapsing remitting MS (RRMS). Yet, the exact mechanisms by which these benefits are achieved remain unknown. CD45RA is a marker for naive T lymphocytes and intercellular adhesion molecule-3 (ICAM-3) is expressed on resting lymphocytes. MATERIAL AND METHODS: Forty-eight patients with RRMS, 24 of them treated with recombinant IFN-beta1b and 24 untreated, were enrolled in this prospective study over 18 months. We investigated the percentage of CD45RA+ ICAM-3+ cells within the total lymphocyte subset in the peripheral blood serially every 3 months and in CSF once at baseline. Detailed clinical examination including Expanded Disability Status Scale (EDSS) score was performed every 3 months and cranial MRI scans were assessed every 6 months. RESULTS: We found a temporary increase in the CD45RA+ ICAM-3+ lymphocyte ratio in peripheral blood of both untreated and IFN beta1b-treated RRMS patients. Moreover, we determined a significant negative correlation (r = -0.5874; P < 0.01) between age as well as the EDSS score (r = 0.3629; P < 0.05) and the percentages of CD45RA+ ICAM-3+ lymphocytes in peripheral blood but a positive correlation between EDSS score and the CD45RA+ ICAM-3+ ratio (r = 0.3913; P < 0.05) in the CSF at baseline. CONCLUSION: CD45RA+ ICAM-3+ lymphocyte ratio in peripheral blood might indicate immunosenescence in MS. However, from our data it cannot be finally concluded whether it is also influenced by IFN-beta1b treatment. PMID- 15527451 TI - Interferon beta-1b modulates serum sVCAM-1 levels in primary progressive multiple sclerosis. AB - Endothelial activation is a key feature of multiple sclerosis (MS) pathogenesis. It is modulated by interferon beta-1b (IFNB-1b) treatment in relapsing-remitting MS (RRMS) patients. This particular pharmacodynamic effect still has to be proven in primary progressive MS (PPMS). In the current study, serum concentrations of soluble vascular cell adhesion molecule-1 (sVCAM-1) and sE-selectin were analyzed longitudinally in 18 PPMS patients before, during and after 12 months of treatment with IFNB-1b. During drug therapy there was a significant early and sustained increase of sVCAM-1 (overall P < 0.0001). Flu-like symptoms induced by IFNB-1b and also concomitant infections were associated with higher sVCAM-1 levels. Neutralizing antibodies to IFNB-1b were associated with lower sVCAM-1 levels. In conclusion, IFNB-1b modulates the adhesion cascade in patients with PPMS in a similar way it does in RRMS. Nevertheless, a clinical effect of IFNB in PPMS still has to be proven in a randomized controlled clinical trial. PMID- 15527452 TI - Cavernous haemangiomas, epilepsy and treatment strategies. AB - BACKGROUND: Cavernous haemangiomas (cavernomas) are benign vascular malformations which can be found at any region within the central nervous system. Epilepsy is the most frequent manifestation of this malformation. Magnetic resonance imaging (MRI) used for the diagnostic evaluation of symptomatic seizures is a sensitive and specific method for the detection of cavernomas. Due to an increased application of MR imaging in the diagnostic evaluation of symptomatic seizures cavernomas have been found with increasing frequency over the past years. To achieve optimal treatment of patients with cavernous haemangiomas and epilepsy both medical and surgical treatment have to be considered. OBJECTIVE: This paper reviews published studies and presents our own results concerning current treatment schedules of epileptogenic cavernomas. RESULTS: In patients with intractable epilpesies surgical treatment of cavernomas seems to be the most successful antiepileptic therapy. Whether microsurgical lesionectomy, including the removal of only the cavernoma, or a more extensive resection is needed to achieve this aim are discussed. CONCLUSION: New data acquired with high resolution imaging (MRI, PET) and electrophysiology indicate that magnetencephalography (MEG) may contribute to the delineation of epileptogenic tissue bordering the lesion, thus permitting appropriate inclusion of this specific area within the resection, instead of performing a simple lesionectomy. PMID- 15527453 TI - The relationship between symptoms and electrophysiological detected compression of the median nerve at the wrist. AB - OBJECTIVE: The aim of this study was to evaluate the relationship between carpal tunnel syndrome symptoms and compression of the median nerve at the wrist in symptomatic patients. METHODS: A total of 250 patients were selected among those referred for electrodiagnostic evaluation with complaints involving hand or wrist. Primary and secondary symptoms were extracted from the answers to the instrument proposed by Levine et al. [J Bone Joint Surg Am 1993;75:1585]. The association of symptoms and the presence of compression of the median nerve at the wrist were ascertained through a multiple logistic regression test. RESULTS: Secondary symptoms (pain and weakness) were inversely associated with the presence of median nerve compression. Furthermore, primary symptoms (paresthesia, disability and nocturnal symptom) occurred similarly in patients with and without electrophysiologic findings of median nerve compression at the wrist. PMID- 15527454 TI - Turns-amplitude analysis of the electromyographic recruitment pattern following upper motor neuron lesions. AB - OBJECTIVE: To assess the pattern of motor unit recruitment of weak muscles in upper motor neuron (UMN) lesions. MATERIALS AND METHODS: Ten patients underwent turns-amplitude analysis (TAA) on the paretic and healthy brachial biceps muscles, in the acute and subacute stages of hemiparesis. The control group comprised 10 age- and sex-matched subjects. RESULTS: Although absent in the acute stage, five patients developed a myogenic cloud pattern in their paretic extremities in the subacute stage; which was statistically significant when compared with controls (P = 0.033). Mean amplitude was reduced in both acute and subacute stages of the hemiparesis compared with controls (P = 0.000). The turns/mean amplitude ratio in the subacute stage was increased compared with both the paretic limbs examined in the acute stage (P = 0.000) and to controls (P = 0.000). CONCLUSION: Abnormalities in the recruitment of motor units in UMN lesions give rise to a myogenic cloud pattern in the TAA, which is prominent in the third month after the initial insult. This may result from the increase in motor unit activity, while the recruitment of bigger motor units is still defective. PMID- 15527455 TI - The outcome of pseudotumor cerebri induced by tetracycline therapy. AB - OBJECTIVE: To demonstrate the association between tetracycline treatment and pseudotumor cerebri (PTC). METHODS: Consecutive patients from two neuro ophthalmic referral centers, who developed PTC syndrome post-treatment with tetracycline, were enrolled and followed for a minimum of 2 years after cessation of tetracycline. RESULTS: A total of 243 consecutive patients were diagnosed with PTC; 18 had concurrent history of tetracycline treatment; a third experienced a limited course of illness with no relapses; 12 had a variable course with a prolonged relapsing illness. Mean duration of tetracycline treatment prior to diagnosis was 2.73 months. CONCLUSIONS: Tetracycline, and especially minocycline, is currently considered a cause or a precipitating factor for PTC. Although there is little information on the natural course of tetracycline induced PTC, the present cases demonstrate that drug withdrawal is curative only in some patients. PMID- 15527456 TI - Descriptions of treatment effects in neurology textbooks recommended for medical students. PMID- 15527460 TI - Review article: impact of night-time reflux on lifestyle - unrecognized issues in reflux disease. AB - Gastro-oesophageal reflux disease (GERD), manifesting with symptoms including heartburn and regurgitation, affects people during both daytime and nocturnal hours. Night-time GERD has been reported to have a greater impact on a patient's life than daytime GERD due to prolonged oesophageal acid exposure time per reflux episode. To further understand this issue, it is important to implement quality of life (QOL) measures. QOL studies are becoming increasingly important to physicians in making clinical decisions, and generic and disease-specific health related QOL (HRQL) tools have been developed to measure a wide variety of topics. There are currently no universally accepted guidelines on how to best measure HRQL in GERD patients. It is important to note that these surveys may not yield accurate results because many GERD sufferers may feel that their symptoms are not serious enough to seek treatment. Some surveys include the GERD-HRQL assessment, the Short Form Health Survey (SF-36), and the Gallup survey. When compared with daytime GERD patients, night-time GERD patients may suffer from sleep deprivation, which in turn leads to physical and emotional problems and a poor overall QOL. Studies indicate that the prevalence and impact of night-time heartburn have been underestimated and that adequate treatment of symptoms is often not achieved. In addition, GERD greatly affects work productivity and leads to a significant economic burden on society. Although limited studies are available on the impact of pharmacological treatment on GERD QOL, recent findings indicate that proton pump inhibitors are more effective than H(2)-receptor antagonists for the improvement of overall QOL. PMID- 15527461 TI - Review article: the pathophysiology of gastro-oesophageal reflux disease - oesophageal manifestations. AB - The pathogenesis of gastro-oesophageal reflux disease (GERD) is multifactorial, involving transient lower oesophageal sphincter relaxations (TLESRs) as well as other lower oesophageal sphincter (LES) pressure abnormalities. GERD is associated with a decrease in LES pressure, which can be provoked by factors such as foods (fat, chocolate, etc.), alcohol, smoking and medications. These factors have also been shown to increase TLESRs. As a result, reflux of acid, bile, pepsin and pancreatic enzymes occurs, leading to oesophageal mucosal injury, which can potentially progress to oesophageal adenocarcinoma in a minority of patients with Barrett's metaplasia. In addition, duodenogastric contents can also contribute to oesophageal injury. Other factors contributing to the pathophysiology of GERD include hiatal hernia, poor oesophageal clearance, delayed gastric emptying and impaired mucosal defensive factors. Hiatal hernia has a permissive role in the pathogenesis of reflux oesophagitis by promoting LES dysfunction. Delayed gastric emptying, resulting in gastric distension, can significantly increase the rate of TLESRs, contributing to postprandial GER. The mucosal defensive factors have an important role in GERD. When excessive acid causes a breakdown in oesophageal epithelial defenses, epithelial resistance may be reduced. Nocturnal GERD is associated with prolonged acid exposure and proximal extent of acid contact, which elevates the risk for oesophageal damage and GERD-related complications. In sum, GERD is a complex problem caused by many factors that are exacerbated when the patient is in the supine position. PMID- 15527462 TI - Review article: supra-oesophageal manifestations of gastro-oesophageal reflux disease and the role of night-time gastro-oesophageal reflux. AB - Gastro-oesophageal reflux disease (GERD) has been associated with a variety of supra-oesophageal symptoms, including asthma, laryngitis, hoarseness, chronic cough, frequent throat clearing and globus pharyngeus. GERD may be overlooked as the underlying mechanism for these symptoms because typical GERD symptoms may be absent, despite abnormal oesophageal acid exposure. Two basic mechanisms linking GERD with laryngeal symptoms have been proposed: direct contact of gastric acid with the upper airway, in some cases due to micro-aspiration, and a vagovagal reflex triggered by acidification of the distal portion of the oesophagus. Gastro oesophageal reflux (GER) during sleep is believed to be an important mechanism for the development of supra-oesophageal complications of GERD, such as asthma and idiopathic pulmonary fibrosis (IPF). Several physiological changes during sleep, including prolonged oesophageal acid contact time, decreased upper oesophageal sphincter pressure, increased gastric acid secretion, decreased salivation, decreased swallowing and a decrease in conscious perception of acid, render an individual more susceptible to reflux-induced injury. Supra-oesophageal symptoms often improve in response to aggressive acid-suppressive therapy. However, many unanswered questions remain regarding the appropriate approach to diagnosis and treatment of patients with GERD-related supra-oesophageal symptoms. In this article we review the relationship between supra-oesophageal symptoms and GERD and, where possible, highlight the evidence supporting the role of night time reflux as a contributing factor to these symptoms. PMID- 15527463 TI - Review article: sleep and its relationship to gastro-oesophageal reflux. AB - Gastro-oesophageal reflux disease (GERD) is among the most common gastrointestinal conditions in the USA. For most symptomatic patients, reflux events occur during both daytime and night-time hours. Whereas daytime reflux events tend to be frequent but brief, reflux events that occur during sleep are comparatively less frequent but significantly longer. Longer oesophageal acid clearance and acid-mucosal contact times during sleep are at least partly due to several physiological changes associated with sleep, including dramatic declines in saliva production and frequency of swallowing, decreased conscious perception of heartburn and consequent arousal and clearance behaviours, and slower gastric emptying. Obstructive sleep apnea syndrome and obesity seem to predispose some patients to nocturnal GERD, and the presence of either of these conditions may help to identify patients with symptoms consistent with GERD. Recognition and treatment of night-time GERD are important because it can be associated with decreased quality of life (including sleep disruption) and increased risk of serious oesophageal and respiratory complications. PMID- 15527464 TI - Review article: oesophageal complications and consequences of persistent gastro oesophageal reflux disease. AB - The major oesophageal complications associated with persistent gastro-oesophageal reflux disease (GERD) include erosive oesophagitis, ulceration, strictures and gastrointestinal (GI) bleeding. Although the causes of these complications are uncertain, studies indicate that erosive oesophagitis may progress to the development of ulcers, strictures and GI bleeding. Pharmacological treatment with proton pump inhibitors is favoured over that with H(2)-receptor antagonists for the treatment of strictures. The treatment of strictures is accomplished with dilation and many favour the concomitant use of proton pump inhibitors. Most gastroenterologists are seeing far fewer oesophageal strictures these days since the introduction of proton pump inhibitors. In addition, research has shown that oesophageal complications have a greater impact on patients suffering from night time GERD than on those suffering from daytime GERD. Barrett's oesophagus is a significant complication associated with persistent GERD and those at risk generally experience a longer duration of symptoms, especially those with a high degree of severity. In addition, there is a strong relationship between Barrett's oesophagus and oesophageal adenocarcinoma. This is in part due to the association of obesity and the development of hiatal hernias. Furthermore, endoscopic screening is being used to detect Barrett's oesophagus and oesophageal adenocarcinoma in persons suffering from chronic GERD, even though screening may not have an impact on outcomes (Sharma P, McQuaid K, Dent J, et al. A critical review of the diagnosis and management of Barrett's esophagus: The AGA Chicago Workshop. Gastroenterology 2004; 127: 310-30.). PMID- 15527465 TI - Review article: diagnosis and management of night-time reflux. AB - Symptoms of gastro-oesophageal reflux disease (GERD) range from mild to severe and, when they occur during night-time hours, can interfere with sleep patterns and reduce overall quality of life. The clinical presentation of GERD is characterized by oesophageal as well as supra-oesophageal symptoms, including otolaryngologic and pulmonary complications. However, GERD may be overlooked as the cause of a patient's supra-oesophageal symptoms because these complaints can occur in the absence of oesophageal symptoms or endoscopic changes. The role of available tools used for GERD diagnosis, including endoscopy, oesophageal pH monitoring and an empirical course of proton pump inhibitor therapy, is discussed. Interventions available to achieve the therapeutic goals of symptom relief and prevention include specific lifestyle modifications and over-the counter as well as prescription pharmacological agents. Patient-initiated, as needed treatment may not be the best choice for managing persistent night-time reflux because it requires patient arousal from sleep. Proton pump inhibitor therapy remains the treatment of choice for patients with more severe symptoms and those with erosive oesophagitis. Few studies have specifically evaluated the role of pharmacological agents in the management of night-time reflux and comparisons are difficult due to the variability in study design and endpoints assessed. PMID- 15527467 TI - The National Service Framework for children, young people and maternity services: developing the evidence base. PMID- 15527468 TI - Facilitators and barriers for co-ordinated multi-agency services. AB - BACKGROUND: Greater collaboration between agencies and the need to improve interagency working is a key policy priority. The lack of co-ordinated multi agency working in children's services has been highlighted in many research studies. Evidence on the facilitators of and barriers to such working and the outcomes for children and families of co-ordinated services is important to inform local developments. METHODS: Literature on multi-agency working was reviewed as part of the evidence gathering to inform the Children's National Service Framework. Searches were mainly concentrated on existing reviews, plus recent studies which included children's services and were not covered by the reviews obtained. RESULTS: There is little evidence on the effectiveness of multi agency working itself or of different models of such working in producing improved outcomes for children and families. However, reviews of evidence on multi-agency working provide consistent findings on facilitators and barriers, including: clear aims, roles and responsibilities and timetables that are agreed between partners; a multi-agency steering group, commitment at all levels of the organizations involved and good systems of communication and information sharing, including IT systems, are central; support and training for staff in new ways of working is needed. There is some evidence that interprofessional programmes of continuing education can help to remove barriers to joint working. CONCLUSIONS: Existing research provides useful information for organizations developing multi agency services. However, there is a need for methodologically sound research which investigates the outcomes of different models of multi-agency working in services for children, includes assessment of cost effectiveness, and explores the ways in which the factors identified as facilitating multi-agency working relate to outcomes. PMID- 15527469 TI - On the road to nowhere? Young disabled people and transition. AB - BACKGROUND: A key issue, and one known to be highly problematic and challenging, concerns the transition of young disabled people from children's services to adult services, and from childhood to adulthood. To inform its recommendations on this issue, the Disabled Children's External Working Group (EWG) for the Children's NSF commissioned a review of research on young disabled people and transition. METHODS: Given the preponderance of grey literature in this area, database searches were supplemented by hand searching of key journals and contacts with key individuals, research units, publishers and funding bodies in order to identify relevant literature on the experiences of transition from the perspectives of young people, parents and practitioners; the outcomes of transition; and evaluations of services provided during this life-stage. RESULTS: The evidence available with regard to transitions of young disabled people provides a clear and consistent picture of the way transitions are currently experienced by young disabled people and their families. There is limited evidence on what works to ensure smooth transitions between services and positive outcomes of transition for the young people themselves. CONCLUSIONS: Existing research is very persuasive on the need to improve transitions for young disabled people. It can also be used to provide clues as to the sorts of support and service configurations that need to be in place to ensure positive transitions. However, further research is needed both at the level of specific service/service model evaluation as well as holistic, longitudinal work to provide evidence on the long-term outcomes of young disabled people as they move into adulthood. PMID- 15527470 TI - Effective services to support children in special circumstances. AB - BACKGROUND: Children living in special circumstances, defined as those who are at risk of achieving poorer outcomes than their peers, have a particular need of good quality, accessible services to promote their health and well-being, yet may be least likely to receive them. AIMS: This review considers the evidence for effective services to support children living in five kinds of special circumstances: (1) those at risk of offending; (2) teenage parents; (3) children whose parents have drug, alcohol or mental health problems; (4) children living with domestic violence; and (5) children who have been abused or neglected. In practice, there is often considerable overlap between these groups, and many children face multiple disadvantage. The review also aims to identify effective strategies for making support services more accessible to hard-to-reach groups. METHODS: Relevant literature was identified through searches of databases and websites, and consultation with experts. Given the broad scope of the topic, the aim was to provide an overview of best evidence rather than to undertake a systematic review. RESULTS: Similar themes underpin the most promising approaches to supporting children in special circumstances. These include: (1) a holistic, multi-agency approach addressing the needs of the whole child rather than compartmentalising children's social, educational, health and care needs; (2) the importance of links between adults' and children's services so that children who are in need because of their parents' circumstances can be identified and supported; and (3) the value of providing children in special circumstances with intensive, targeted support within a framework of universal provision. CONCLUSIONS: Overall, there is a lack of well-designed evaluations of the effectiveness of UK services and programmes for children in special circumstances. The existing evidence base would be strengthened by the development of better outcome measures, by exploring the characteristics of effective services from the perspective of different stakeholders (including children and young people themselves) and by extending the current problem oriented approach to consider the factors that promote resilience and coping. PMID- 15527471 TI - Improving services for disadvantaged childbearing women. AB - BACKGROUND: Tackling health inequalities is high on the political agenda of the Labour government. The government wants to reduce inequalities through policies based on evidence, the health of childbearing women and their babies being priority areas for action. National Service Frameworks (NSFs) are being set up to ensure high quality services for all groups. The External Working Group looking into maternity services for the Children's NSF seeks evidence upon which to plan strategies for all women, while focusing attention on the most disadvantaged. Wide differentials are noted between high- and low-income groups in the outcomes of pregnancy and the health of babies. The worst outcomes appear to be concentrated in small subgroups of disadvantaged women. AIM: To report on a review of studies of interventions improving perinatal outcomes for disadvantaged women, to inform policy and practice around the organization and delivery of statutory services in the UK. METHOD: We searched six relevant databases for reviews, intervention studies, audits and descriptions of services reporting outcomes for disadvantaged groups of women, published between 1990 and 2003. Synthesis was performed around what works and what does not work. Gaps in the evidence base were identified. RESULTS: We found limited evidence of effective and promising interventions for childbearing women from minority ethnic groups, women experiencing domestic violence, women with mental health illness and HIV infected women. Few studies were well-designed or powered to detect effectiveness. There were no studies of interventions for women prisoners, homeless women and travellers. CONCLUSIONS: Searching for intervention studies primarily by participant subgroup has brought in evidence from few well-designed studies on which to plan policy. Combining this approach with searching for intervention studies addressing behaviour like smoking, and needs like social support, may provide further evidence to tackle inequalities in the perinatal period. PMID- 15527472 TI - The participation of children and young people in decisions about UK service development. AB - BACKGROUND: The involvement of children and young people in decisions regarding service development is well supported in government policy and underpinned by the UN Convention on the Rights of the Child. Information on the extent, nature and outcomes of children and young people's participation can inform further development in this area. METHODS: Systematic literature searches, plus contact with professional networks, were used to gather and review evidence on children and young people's participation. RESULTS: There is a rapidly developing body of information describing and analysing innovative practices in this field. However, there is also a smaller, but substantial, amount of evidence demonstrating the limited extent of current involvement. A good deal of guidance is now available about how to promote the involvement of children and young people. However, the basis of this advice is not always clear, and more evidence about children's views and their experience of participation in public decision-making is required. Issues identified as barriers to change included adult attitudes and intransigence, lack of training for key adults, lack of clarity leading to tokenism, the nature of organizations (i.e. their formality, complexity, bureaucracy and internal politics) and the short-term nature of much funding. The evidence suggests that good practice includes a listening culture among staff, clarity, flexibility, adequate resources, skills development and training for staff and participating children and young people, inclusion of marginalized groups, feedback and evaluation. There is only limited evidence that children and young people's involvement in public decision-making leads to more appropriate services, although there is evidence that participating children and young people benefit in terms of personal development and that staff and organizations learn more about their views. CONCLUSIONS: The value of the participation of children and young people in public decision-making is now well accepted, and is recognized in the standards set in the Children's National Service Framework. However, there is an urgent need for internal and external evaluations of children's involvement. PMID- 15527473 TI - Promoting and protecting the health of children and young people. AB - BACKGROUND: The health-related behaviours adopted by children and young people can have both immediate and long-term health effects. Health promotion interventions that target children and young people can lay the foundations of a healthy lifestyle that may be sustained into adulthood. This paper is based on a selective review of evidence relating to health promotion in childhood, carried out to support the external working group on the 'Healthy Child' module of the Children's National Service Framework. METHODS: This is a selective review of mainly secondary research. It focuses on injury prevention, support for parenting and the promotion of good mental health, and promoting a healthy diet and physical activity amongst children and young people. FINDINGS: In many areas, the quality of primary research into health promotion interventions aimed at children and young people is poor. Interventions are heterogeneous and not described in sufficient detail. Sample sizes tend to be small, and there are commonly problems of bias. Despite these difficulties, there is good evidence for a range of interventions, including (1) area road safety schemes; (2) combining a variety of approaches to the promotion of the use of safety equipment, including legislation and enforcement, loan/assisted purchase/giveaway schemes, education, fitting and maintenance of safety equipment; (3) school-based mental health promotion; (4) parenting support; (5) interventions that promote and facilitate 'lifestyle' activity for children, such as walking and cycling to school, and those that aim to reduce sedentary behaviours such as parent education to reduce the time children spend watching TV and using computers; and (6) controlling advertising of unhealthy food that is aimed at children. CONCLUSIONS: There are effective interventions to promote and protect the health of children and young people that require action across the five areas described in the Ottawa Charter. Health, social care and education services have a direct role in the delivery of many of these interventions and, in other areas, a role in collaborative work with other agencies, in lobbying for policy change and in raising the profile of child health promotion. Further research is needed using larger study populations, and closely defined interventions, both targeted and universal, in order to fill some of the current gaps in the evidence base for health promotion in children and young people. PMID- 15527474 TI - Psycho-educational interventions for children with chronic disease, parents and siblings: an overview of the research evidence base. AB - BACKGROUND: The role of psycho-educational interventions in facilitating adaptation to chronic disease has received growing recognition and is in keeping with policy developments advocating greater involvement of patients in their own care. The purpose of this paper is to provide an overview of the current literature regarding the effectiveness of psycho-educational interventions for children and adolescents with chronic disease, their parents and siblings. METHODS: Electronic searches were conducted using AMED, CINAHL, Cochrane Database, DARE, HTA, MEDLINE, NHS EED, PsycLIT, PsycINFO, and PubMED. Inclusion criteria were systematic reviews, meta-analyses and overviews based on traditional reviews of published literature. The titles of papers were reviewed, abstracts were obtained and reviewed, and full copies of selected papers were obtained. RESULTS: No reviews of psycho-educational interventions were found for either parents or siblings. Twelve reviews of interventions for children and adolescents were identified: chronic disease in general (three); chronic pain (one); asthma (three); chronic fatigue syndrome (CFS)/myalgic encephalomyelitis (ME) (one); diabetes (two); juvenile idiopathic arthritis (JIA) (one) and one informational intervention for paediatric cancer patients. The main focus was on disease management (particularly in asthma and diabetes) with less attention being paid to psychosocial aspects of life with a chronic condition. Overall, there is evidence of effectiveness for interventions incorporating cognitive behavioural techniques on variables such as self-efficacy, self-management of disease, family functioning, psychosocial well-being, reduced isolation, social competence, knowledge, hope, pain (for chronic headache), lung function (asthma), days absent from school (asthma), visits to A & E (asthma), fatigue (CFS), and metabolic control (diabetes). A number of gaps and limitations were identified across all disease categories, such as inadequate description of interventions, small sample sizes, and lack of evidence regarding cost-effectiveness. CONCLUSION: This overview has highlighted the need to extend the evidence base for psycho-educational interventions, particularly in a UK context. It is essential that effective interventions are implemented and embedded in service provision in order to maximize empowerment through self-care for children, adolescents and their parents. PMID- 15527475 TI - A literature review to identify interventions to improve the use of medicines in children. AB - BACKGROUND: It is estimated that 200 million prescriptions for children and adolescents were issued in the UK during 2002. Therefore, it is important for the National Service Framework for Children (NSFC) to include advice on managing medicines effectively for children. This literature review was performed at the request of the NSFC Medicines External Working Group in order to provide underpinning evidence in the development of advice on managing medicines. METHODS: Detabases, websites and conference abstracts were searched systematically to identify information on managing medicines in children in 2003. This article reported the results on medication review, concordance, enhanced medicines access through community pharmacy services and the use of medicines in schools. RESULTS AND CONCLUSIONS: Although there is little evidence specific to paediatrics, the objectives and rationale of medication review could be expected to apply to chronic diseases in children. Issues such as polypharmacy, wastage, repeat prescriptions and medication problems could be similar. The benefits seen in adults may also occur in children, and medication review may possibly have a role in the management of medicines in children. There is an obvious role for pharmacists in ensuring the safety of over-the-counter medications and provision of information and education to parents, carers and adolescents. Evaluation and provision of necessary education and training to community pharmacists is needed, even in the most basic paediatric issues such as sugar-free medications. The evidence suggests that treatment compliance and adherence are generally lower in children than in adults, particularly in adolescents as they approach independence. Those with learning disabilities and infants are likely to be at risk of non-compliance, although little work has been done in these populations. Children and adolescents need appropriate parental and professional support in taking control of their medication and treatment. The management of medicines in school would appear to be far from ideal. Further research into school-based medicines education and outreach clinics would also be beneficial. PMID- 15527476 TI - Child public health and social welfare: lessons from the evidence. PMID- 15527477 TI - Childhood disadvantage and health inequalities: a framework for policy based on lifecourse research. AB - Tackling health inequalities is moving up the policy agenda of richer societies like the UK, with governments looking for evidence to guide policy review and development. Observational studies of how childhood disadvantage compromises health in adulthood are an important part of the evidence base, but are largely inaccessible to the policy community. We develop a framework which captures the findings of these studies. Our framework highlights how disadvantage in childhood adversely affects both socio-economic circumstances and health in adulthood through a set of interlocking processes. Key among these are children's developmental health (their physical, cognitive and emotional development) and health behaviours, together with the associated educational and social trajectories. In breaking down the link between childhood disadvantage and adult health into its constituent elements, the framework provides a basis for understanding where and how policies can make a difference. The paper argues that the process of policy review and development needs to include both new programmes and the mainstream policies in which they are embedded. PMID- 15527478 TI - Growing up in disadvantage: a systematic review of the qualitative evidence. AB - BACKGROUND AND OBJECTIVES: Childhood disadvantage has lasting negative effects on children's health and well-being. However, the impact of poverty and associated disadvantage on children's lives as children is under-researched. This paper reviews a small, but important, group of qualitative studies that prioritize children's perspectives on growing up in disadvantage, exploring the social resources that they typically draw upon. METHODS: Systematic review methods were used to locate and appraise qualitative studies that take as their focus children's subjective accounts of living in disadvantage. Data from the studies were then synthesized using meta-ethnographic methods. RESULTS: Children and young people describe aspects of family relationships, friendships and neighbourhoods that help to mitigate the impact of disadvantage on their well being. However, their accounts demonstrate that such resources are not always and unambignously experienced as supportive and protective. DISCUSSION: This systematic review highlights the value of social resources available to children living in poor circumstances, but also points up their limitations. Poverty, by its very nature, compromises the worth of these resources. PMID- 15527479 TI - The impact of day care on socially disadvantaged families: an example of the use of process evaluation within a randomized controlled trial. AB - AIM: This paper describes a process evaluation that was conducted alongside a randomized controlled trial of out-of-home pre-school day care. The evaluation aimed to: (1) describe the intervention; (2) document the day care received by participating families; (3) describe the social context of the trial; and (4) provide data to assist in the interpretation of trial outcomes. METHODS: The setting for the trial was an out-of-home day care Centre in Hackney, East London. Process data were collected through the use of questionnaires, interviews, and researcher field-notes. Data from questionnaires were collected from 120 mothers and included data on 143 children. Interviews were undertaken with 21 participating mothers. Staff also completed questionnaires and the Head of the Centre was interviewed. The quality of care provided was assessed using the Early Childhood Environment Rating Scale. RESULTS: Process data collected during the trial suggest that the day care provided was education-led, flexible in catering to families' needs, and was of a very high quality. The social context of the trial resulted in financial pressures, which may well have influenced the intervention provided. Data collected through in-depth interviews suggested that it may be the flexibility of day care that is particularly important in allowing women to return to paid employment, but that the loss of benefits when starting work may have meant no increase in household income. CONCLUSION: The paper illustrates the value of conducting a process evaluation alongside a randomized trial, particularly where complex interventions are involved. In this case, where the intervention was not provided by the research team, the evaluation allowed an insight into the content of a multifaceted intervention, which is useful in interpreting the trial's results, and in explaining the possible effects of the social context on the intervention. PMID- 15527480 TI - The effect of income inequality and macro-level social policy on infant mortality and low birthweight in developed countries--a preliminary systematic review. AB - OBJECTIVE: To carry out a preliminary systematic review of literature to address the question - among rich nations (or states within nations) what is the evidence that income inequality and differences in macro-level social policy affect rates of infant mortality and low birthweight (LBW)? STUDY DESIGN: A systematic literature review. SEARCH STRATEGY: Medline database (1968-August 2003) was searched for empirical studies of the relationship between macro-level economic and social policies in rich nations and rates of infant mortality (IMR) and LBW. Cross-national comparison of infant mortality and LBW that did not compare the effects of macro-level economic and social policies was excluded from the review as were studies including less developed countries. Keywords representing IMR and LBW were entered into Medline along with exposures related to international comparison and macro-level policy. Abstracts obtained from the initial search were reviewed for relevant studies. Full papers of potentially relevant studies were obtained and reviewed for inclusion. Secondary search of papers cited in included papers was undertaken. For this review, papers were not excluded on the basis of quality although methodological limitations were commented on and taken into account in interpreting the results. Summary statistics were not estimated. RESULTS: Twelve studies, fulfilling the inclusion criteria, were identified. Ten studies examined the association of IMR with income inequality, eight of which reported a statistically significant positive association with higher levels of inequality after adjustment for a range of variables. Six studies reported significant positive associations of IMR with other indicators of less re distributive social and economic policy. Associations with LBW were reported in four studies; three showed significant positive associations with higher levels of income inequality and one showed no association with low levels of parental leave entitlement. Methodological differences, particularly the wide range of variables used to adjust for confounding, make interpretation of the findings difficult. CONCLUSIONS: The results of this review represent a preliminary attempt to summarize the literature linking macro-level economic and social policies in rich nations with IMR and LBW. The findings, taking account of the methodological limitations of the review and of the included studies, suggest a statistically significant association between IMR and higher income inequality and other indicators of less re-distributive social policy. Only three studies examined the association of income inequality with LBW and, although they suggest a significant association, further studies will be needed to confirm this finding. PMID- 15527481 TI - The social and economic consequences of childhood asthma across the lifecourse: a systematic review. AB - BACKGROUND: Asthma prevalence rates are high, and may be increasing in the Western world, particularly among children. The aim of this study was to evaluate the longer-term social and economic consequences of having asthma as a child and to determine whether adverse consequences are more severe for poorer children. METHODS: Results from published and unpublished, quantitative and qualitative studies were synthesized narratively to examine the impact of childhood-onset asthma on school attendance, academic achievement and employment in adulthood. The question of whether the impact differed for different social groups was also examined. FINDINGS: Twenty-nine good quality studies were identified, including in total 12 183 children with asthma or wheeze. Compared with asymptomatic children, those with asthma missed more days of school (additional absence as a result of asthma ranged from 2.1 to 14.8 days). Studies of academic achievement found that children with asthma performed as well as their healthy peers. The existing evidence on labour market performance is sparse, but there is an indication that people with asthma during childhood experience disadvantage in later employment. In an examination of consequences by social position, children with asthma from deprived areas were more likely to miss school than their more affluent peers, and minority ethnic children were also more likely to have poor school attendance. The only qualitative study suggested that children with asthma strove to participate fully in every aspect of their daily lives. INTERPRETATION: Although asthma limits children's daily activities and affects their social activities, this research synthesis found little evidence of major, adverse long term social and economic consequences in studies reviewed. Further longitudinal research using comparison groups is needed to fill key gaps in the existing evidence base. PMID- 15527491 TI - Migration through host cells: the first steps of Plasmodium sporozoites in the mammalian host. AB - Malaria starts with the infection of the liver by Plasmodium sporozoites. This form of the parasite migrates through several host cells breaching their plasma membranes before infecting a final hepatocyte which they enter forming a parasitophorous vacuole. It is still controversial why Plasmodium sporozoites migrate through host cells. By reviewing the most recent literature, we hope to give an insight on the different steps of host invasion in which migration through cells is involved and on the possible role for this mechanism in infection. PMID- 15527492 TI - Liver invasion by malarial parasites--how do malarial parasites break through the host barrier? AB - Malarial transmission to the human host is established by sporozoite infection of the liver. Sporozoites are released from the mosquito salivary glands and carried by the blood flow to the liver sinusoid. In the sinusoid, sporozoites leave the blood circulation by crossing the sinusoidal cell layer to infect hepatocytes, the site for their development into the erythrocyte-invasive forms. Traversal of the sinusoidal cell layer and subsequent hepatocyte infection are the most important events in sporozoite liver invasion, but the molecular basis of both events remains to be elucidated. The present review of sporozoite liver invasion focuses on recent advances in this topic obtained by application of reverse genetics. Sporozoites traverse host cells, rupturing the host cell membrane in the process. Three microneme proteins have important roles in this motility. Disruption of one of these genes abolishes or severely impairs cell traversal without affecting other types of invasive motility. Studies using these disruptant parasites indicate that cell-traversal ability is required for crossing the sinusoidal cell layer and accessing the hepatocytes for infection. This process is homologous to midgut epithelium penetration by the malarial ookinete, because identical or paralogous genes are critically involved in both processes. After arrival at the hepatocyte, the invasion mode of the sporozoites switches from cell traversal to hepatocyte infection. PMID- 15527493 TI - Burkholderia cepacia complex isolates survive intracellularly without replication within acidic vacuoles of Acanthamoeba polyphaga. AB - We have previously demonstrated that isolates of the Burkholderia cepacia complex can survive intracellularly in murine macrophages and in free-living Acanthamoeba. In this work, we show that the clinical isolates B. vietnamiensis strain CEP040 and B. cenocepacia H111 survived but did not replicate within vacuoles of A. polyphaga. B. cepacia-containing vacuoles accumulated the fluid phase marker Lysosensor Blue and displayed strong blue fluorescence, indicating that they had low pH. In contrast, the majority of intracellular bacteria within amoebae treated with the V-ATPse inhibitor bafilomycin A1 localized in vacuoles that did not fluoresce with Lysosensor Blue. Experiments using bacteria fluorescently labelled with chloromethylfluorescein diacetate demonstrated that intracellular bacteria remained viable for at least 24 h. In contrast, Escherichia coli did not survive within amoebae after 2 h post infection. Furthermore, intracellular B. vietnamiensis CEP040 retained green fluorescent protein within the bacterial cytoplasm, while this protein rapidly escaped from the cytosol of phagocytized heat-killed bacteria into the vacuolar lumen. Transmission electron microscopy analysis confirmed that intracellular Burkholderia cells were structurally intact. In addition, both Legionella pneumophila- and B. vietnamiensis-containing vacuoles did not accumulate cationized ferritin, a compound that localizes within the lysosome. Thus, our observations support the notion that B. cepacia complex isolates can use amoebae as a reservoir in the environment by surviving without intracellular replication within an acidic vacuole that is distinct from the lysosomal compartment. PMID- 15527494 TI - NO means 'yes' in the squid-vibrio symbiosis: nitric oxide (NO) during the initial stages of a beneficial association. AB - During colonization of the Euprymna scolopes light organ, symbiotic Vibrio fischeri cells aggregate in mucus secreted by a superficial ciliated host epithelium near the sites of eventual inoculation. Once aggregated, symbiont cells migrate through ducts into epithelium-lined crypts, where they form a persistent association with the host. In this study, we provide evidence that nitric oxide synthase (NOS) and its product nitric oxide (NO) are active during the colonization of host tissues by V. fischeri. NADPH-diaphorase staining and immunocytochemistry detected NOS, and the fluorochrome diaminofluorescein (DAF) detected its product NO in high concentrations in the epithelia of the superficial ciliated fields, ducts, and crypt antechambers. In addition, both NOS and NO were detected in vesicles within the secreted mucus where the symbionts aggregate. In the presence of NO scavengers, cells of a non-symbiotic Vibrio species formed unusually large aggregates outside of the light organ, but these bacteria did not colonize host tissues. In contrast, V. fischeri effectively colonized the crypts and irreversibly attenuated the NOS and NO signals in the ducts and crypt antechambers. These data provide evidence that NO production, a defense response of animal cells to bacterial pathogens, plays a role in the interactions between a host and its beneficial bacterial partner during the initiation of symbiotic colonization. PMID- 15527495 TI - Interaction of Neisseria meningitidis with human brain microvascular endothelial cells: role of MAP- and tyrosine kinases in invasion and inflammatory cytokine release. AB - Neisseria meningitidis traversal across the blood-cerebrospinal fluid barrier is an essential step in the pathogenesis of bacterial meningitis. We have previously shown that invasion of human brain microvascular endothelial cells (HBMEC) by meningococci is mediated by bacterial outer membrane protein Opc that binds fibronectin, thereby anchoring the bacterium to the integrin alpha 5 beta 1 receptor on the endothelial cell surface. However, subsequent signal transduction mechanisms essential for or regulated by N. meningitidis adhesion and invasion, or HBMEC responses to N. meningitidis are unknown. In this report we investigated the role of c-Jun N-terminal kinases 1 and 2 (JNK1 and JNK2), p38 mitogen activated (MAP) kinase and protein tyrosine kinases in endothelial-N. meningitidis interaction. Binding of meningococci to HBMEC phosphorylated and activated JNK1 and JNK2 and p38 MAPK as well as their direct substrates c-Jun and MAP kinase activated kinase-2 (MAPKAPK-2), respectively. Non-invasive meningococcal strains lacking opc gene (opc mutants and sequence type 11 complex meningococci) still activated p38 MAPK, however, failed to activate JNK. Inhibition of JNK1 and JNK2 significantly reduced internalization of N. meningitidis by HBMEC without affecting its adherence. Blocking the endothelial integrin alpha 5 beta 1 also decreased N. meningitidis-induced JNK activation in HBMEC. These findings indicate the crucial role of JNK signalling pathway in N. meningitidis invasion in HBMEC. In contrast, p38 MAPK pathway was important for the control of interleukin-6 (IL-6) and IL-8 release by HBMEC. Genistein, a protein tyrosine kinase inhibitor, decreased both invasion of N. meningitidis into HBMEC and IL-6 and IL-8 release, indicating that protein tyrosine kinases, which link signals from integrins to intracellular signalling pathways are essential for both bacterial internalization and cytokine secretion by HBMEC. PMID- 15527496 TI - TccP is an enterohaemorrhagic Escherichia coli O157:H7 type III effector protein that couples Tir to the actin-cytoskeleton. AB - Subversion of host cell actin microfilaments is the hallmark of enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) Escherichia coli infections. Both pathogens translocate the trans-membrane receptor protein translocated intimin receptor (Tir), which links the extracellular bacterium to the cell cytoskeleton. While both converge on neural Wiskott-Aldrich syndrome protein (N-WASP), Tir-mediated actin accretion by EPEC and EHEC differ in that Tir(EPEC) requires both tyrosine phosphorylation and the host adaptor protein Nck, whereas Tir(EHEC) is not phosphorylated and utilizes an unidentified linker. Here we report the identification of Tir-cytoskeleton coupling protein (TccP), a novel EHEC effector that displays an Nck-like coupling activity following translocation into host cells. A tccP mutant did not affect Tir translocation and focusing but failed to recruit alpha-actinin, Arp3, N-WASP and actin to the site of bacterial adhesion. When expressed in EPEC, bacterial-derived TccP restored actin polymerization activity following infection of an Nck-deficient cell line. TccP has a similar biological activity on infected human intestinal explants ex vivo. Purified TccP activates N-WASP stimulating, in the presence of Arp2/3, actin polymerization in vitro. These results show that EHEC translocates both its own receptor (Tir) and an Nck-like protein (TccP) to facilitate actin polymerization. PMID- 15527497 TI - Lipoprotein lipase in hemodialysis patients: indications that low molecular weight heparin depletes functional stores, despite low plasma levels of the enzyme. AB - BACKGROUND: Lipoprotein lipase (LPL) has a central role in the catabolism of triglyceride-rich lipoproteins. The enzyme is anchored to the vascular endothelium through interaction with heparan sulphate proteoglycans and is displaced from this interaction by heparin. When heparin is infused, there is a peak of LPL activity accompanied by a reduction in triglycerides (TG) during the first hour, followed by a decrease in LPL activity to a stable plateau during the remaining session while TG increase towards and beyond baseline. This suggests that tissue stores of LPL become depleted. It has been argued that low molecular weight (LMW) heparins cause less disturbance of the LPL system than conventional heparin does. METHODS: We have followed LPL activity and TG during a dialysis session with a LMW heparin (dalteparin) using the same patients and regime as in a previous study with conventional heparin, i.e. a primed infusion. RESULTS: The shape of the curve for LPL activity resembled that during the earlier dialyses with conventional heparin, but the values were lower during dialysis with dalteparin. The area under the curve for LPL activity during the peak period (0 180 minutes) was only 27% and for the plateau period (180-240 minutes) it was only 36% of that observed with conventional heparin (p < 0.01). These remarkably low plasma LPL activities prompted us to re-analyze LPL activity and to measure LPL mass in frozen samples from our earlier studies. There was excellent correlation between the new and old values which rules out the possibility of assay variations as a confounding factor. TG increased from 2.14 mmol/L before, to 2.59 mmol/L after the dialysis (p < 0.01). From 30 minutes on, the TG values were significantly higher after dalteparin compared to conventional heparin (p < 0.05). CONCLUSION: These results indicate that LMW heparins disturb the LPL system as much or more than conventional heparin does. PMID- 15527498 TI - Low birth weight and longitudinal trends of cardiovascular risk factor variables from childhood to adolescence: the bogalusa heart study. AB - BACKGROUND: Several studies have linked low birth weight to adverse levels of cardiovascular risk factors and related diseases. However, information is sparse at a community level in the U.S. general population regarding the effects of low birth weight on the longitudinal trends in cardiovascular risk factor variables measured concurrently from childhood to adolescence. METHODS: Longitudinal analysis was performed retrospectively on data collected from the Bogalusa Heart Study cohort (n = 1141; 57% white, 43% black) followed from childhood to adolescence by repeated surveys between 1973 and 1996. Subjects were categorized into low birth weight (below the race-specific 10th percentile; n = 123) and control (between race-specific 50-75th percentile; n = 296) groups. RESULTS: Low birth weight group vs control group had lower mean HDL cholesterol (p = 0.05) and higher LDL cholesterol (p = 0.05) during childhood (ages 4-11 years); higher glucose (p = 0.02) during adolescence. Yearly rates of change from childhood to adolescence in systolic blood pressure (p = 0.02), LDL cholesterol (p = 0.05), and glucose (p = 0.07) were faster, and body mass index (p = 0.03) slower among the low birth weight group. In a multivariate analysis, low birth weight was related independently and adversely to longitudinal trends in systolic blood pressure (p = 0.004), triglycerides (p = 0.03), and glucose (p = 0.07), regardless of race or gender. These adverse associations became amplified with age. CONCLUSIONS: Low birth weight is characterized by adverse developmental trends in metabolic and hemodynamic variables during childhood and adolescence; and thus, it may be an early risk factor in this regard. PMID- 15527499 TI - The use of Open Reading frame ESTs (ORESTES) for analysis of the honey bee transcriptome. AB - BACKGROUND: The ongoing efforts to sequence the honey bee genome require additional initiatives to define its transcriptome. Towards this end, we employed the Open Reading frame ESTs (ORESTES) strategy to generate profiles for the life cycle of Apis mellifera workers. RESULTS: Of the 5,021 ORESTES, 35.2% matched with previously deposited Apis ESTs. The analysis of the remaining sequences defined a set of putative orthologs whose majority had their best-match hits with Anopheles and Drosophila genes. CAP3 assembly of the Apis ORESTES with the already existing 15,500 Apis ESTs generated 3,408 contigs. BLASTX comparison of these contigs with protein sets of organisms representing distinct phylogenetic clades revealed a total of 1,629 contigs that Apis mellifera shares with different taxa. Most (41%) represent genes that are in common to all taxa, another 21% are shared between metazoans (Bilateria), and 16% are shared only within the Insecta clade. A set of 23 putative genes presented a best match with human genes, many of which encode factors related to cell signaling/signal transduction. 1,779 contigs (52%) did not match any known sequence. Applying a correction factor deduced from a parallel analysis performed with Drosophila melanogaster ORESTES, we estimate that approximately half of these no-match ESTs contigs (22%) should represent Apis-specific genes. CONCLUSIONS: The versatile and cost-efficient ORESTES approach produced minilibraries for honey bee life cycle stages. Such information on central gene regions contributes to genome annotation and also lends itself to cross-transcriptome comparisons to reveal evolutionary trends in insect genomes. PMID- 15527500 TI - Snoring in primary school children and domestic environment: a Perth school based study. AB - BACKGROUND: The home is the predominant environment for exposure to many environmental irritants such as air pollutants and allergens. Exposure to common indoor irritants including volatile organic compounds, formaldehyde and nitrogen dioxide, may increase the risk of snoring for children. The aim of this study was to investigate domestic environmental factors associated with snoring in children. METHODS: A school-based respiratory survey was administered during March and April of 2002. Nine hundred and ninety six children from four primary schools within the Perth metropolitan area were recruited for the study. A sub group of 88 children aged 4-6 years were further selected from this sample for domestic air pollutant assessment. RESULTS: The prevalences of infrequent snoring and habitual snoring in primary school children were 24.9% and 15.2% respectively. Passive smoking was found to be a significant risk factor for habitual snoring (odds ratio (OR) = 1.77; 95% confidence interval (CI): 1.20 2.61), while having pets at home appeared to be protective against habitual snoring (OR = 0.58; 95% CI: 0.37-0.92). Domestic pollutant assessments showed that the prevalence of snoring was significantly associated with exposure to nitrogen dioxide during winter. Relative to the low exposure category (<30 microg/m3), the adjusted ORs of snoring by children with medium (30 - 60 microg/m3) and high exposures (> 60 microg/m3) to NO2 were 2.5 (95% CI: 0.7-8.7) and 4.5 (95% CI: 1.4-14.3) respectively. The corresponding linear dose-response trend was also significant (P = 0.011). CONCLUSION: Snoring is common in primary school children. Domestic environments may play a significant role in the increased prevalence of snoring. Exposure to nitrogen dioxide in domestic environment is associated with snoring in children. PMID- 15527501 TI - Selective inhibition of c-Myb DNA-binding by RNA polymers. AB - BACKGROUND: The transcription factor c-Myb is expressed in hematopoietic progenitor cells and other rapidly proliferating tissues, regulating genes important for proliferation, differentiation and survival. The DNA-binding domain (DBD) of c-Myb contains three tandemly arranged imperfect repeats, designated Myb domain R1, R2 and R3. The three-dimensional structure of the DBD shows that only the second and third Myb domains are directly involved in sequence-specific DNA binding, while the R1 repeat does not contact DNA and only marginally affects DNA binding properties. No structural information is available on the N-terminal 30 residues. Since deletion of the N-terminal region including R1 plays an important role in oncogenic activation of c-Myb, we asked whether this region confers properties beyond DNA-binding to the neighbouring c-Myb DBD. RESULTS: Analysis of a putative RNA-binding function of c-Myb DBD revealed that poly(G) preferentially inhibited c-Myb DNA-binding. A strong sequence-selectivity was observed when different RNA polymers were compared. Most interesting, the poly(G) sensitivity was significantly larger for a protein containing the N-terminus and the R1 repeat than for the minimal DNA-binding domain. CONCLUSION: Preferential inhibition of c-Myb DNA binding by poly(G) RNA suggests that c-Myb is able to interact with RNA in a sequence-selective manner. While R2 and R3, but not R1, are necessary for DNA-binding, R1 seems to have a distinct role in enhancing the RNA-sensitivity of c-Myb. PMID- 15527503 TI - Validating the Time and Change test to screen for dementia in elderly Koreans. AB - BACKGROUND: We assessed the applicability of the T&C test as an accurate and convenient means to screen for dementia in primary care and community settings. METHODS: The study group comprised 59 patients and 405 community participants, all of who were aged 65 years and over. The time component of the T&C test evaluated the ability of a subject to comprehend clock hands that indicated a time of 11:10, while the change component of the T&C test evaluated the ability of a subject to make 1,000 Won from a group of coins with smaller denominations (one 500, seven 100, and seven 50 Won coins). RESULTS: The T&C test had a sensitivity and specificity of 73.0 and 90.9%, respectively, and positive and negative predictive values of 93.1, and 66.7%, respectively. The test-retest and interobserver agreement rates were both 95% (kappa = 0.91) (time interval, 24 hours). The association between the T&C test and K-MMSE test was modest, while significant (r = 0.422, p < 0.001). The T&C test scores were not influenced by educational status. CONCLUSIONS: We conclude that the T&C test is useful as supplemental testing of important domains (e.g., calculation, conceptualization, visuospatial) to traditional measures such as the MMSE. However, because T&C test is simple, rapid, and easy to use, it can be applied conveniently to elderly subjects by non-specialist personnel who receive training. PMID- 15527504 TI - Treatment outcomes in locally advanced colorectal carcinoma. AB - BACKGROUND: Locally advanced colorectal cancers form a distinct subgroup where contiguous organs could be involved without distant metastases and so may be amenable to curative surgical resection. It was our objective to report our experience in treating six such patients with operable locally advanced colorectal carcinomas. METHODS: We retrospectively reviewed the case notes of 47 patients who were diagnosed with colorectal cancers at M S Ramaiah Medical Teaching Hospital between the years 1996 - 2001. Six patients were identified with T4 lesions, adjacent organ involvement and with no nodal involvement. The treatments and outcomes for these patients were then reviewed. RESULTS: Two of three patients with rectal malignancies who underwent pelvic exenteration succumbed to disease recurrence within the first 18 months. One of the three patients with colonic cancers died of non malignant causes. The other two are disease free till date. CONCLUSIONS: Aggressive multivisceral resections for locally advanced colonic cancers might be appropriate. Rectal cancers when locally advanced may be considered for pelvic exenteration, but a more guarded prognosis may apply. PMID- 15527502 TI - Psychiatric diagnoses in 3275 suicides: a meta-analysis. AB - BACKGROUND: It is well known that most suicide cases meet criteria for a psychiatric disorder. However, rates of specific disorders vary considerably between studies and little information is known about gender and geographic differences. This study provides overall rates of total and specific psychiatric disorders in suicide completers and presents evidence supporting gender and geographic differences in their relative proportion. METHODS: We carried out a review of studies in which psychological autopsy studies of suicide completers were performed. Studies were identified by means of MEDLINE database searches and by scanning the reference list of relevant publications. Twenty-three variables were defined, 16 of which evaluating psychiatric disorders. Mantel-Haenszel Weighted Odds Ratios were estimated for these 16 outcome variables. RESULTS: Twenty-seven studies comprising 3275 suicides were included, of which, 87.3% (SD 10.0%) had been diagnosed with a mental disorder prior to their death. There were major gender differences. Diagnoses of substance-related problems (OR = 3.58; 95% CI: 2.78-4.61), personality disorders (OR = 2.01; 95% CI: 1.38-2.95) and childhood disorders (OR = 4.95; 95% CI: 2.69-9.31) were more common among male suicides, whereas affective disorders (OR = 0.66; 95% CI: 0.53-0.83), including depressive disorders (OR = 0.53; 95% CI: 0.42-0.68) were less common among males. Geographical differences are also likely to be present in the relative proportion of psychiatric diagnoses among suicides. CONCLUSIONS: Although psychopathology clearly mediates suicide risk, gender and geographical differences seem to exist in the relative proportion of the specific psychiatric disorders found among suicide completers. PMID- 15527505 TI - On the potential role of glutamate transport in mental fatigue. AB - Mental fatigue, with decreased concentration capacity, is common in neuroinflammatory and neurodegenerative diseases, often appearing prior to other major mental or physical neurological symptoms. Mental fatigue also makes rehabilitation more difficult after a stroke, brain trauma, meningitis or encephalitis. As increased levels of proinflammatory cytokines are reported in these disorders, we wanted to explore whether or not proinflammatory cytokines could induce mental fatigue, and if so, by what mechanisms.It is well known that proinflammatory cytokines are increased in major depression, "sickness behavior" and sleep deprivation, which are all disorders associated with mental fatigue. Furthermore, an influence by specific proinflammatory cytokines, such as interleukin (IL)-1, on learning and memory capacities has been observed in several experimental systems. As glutamate signaling is crucial for information intake and processing within the brain, and due to the pivotal role for glutamate in brain metabolism, dynamic alterations in glutamate transmission could be of pathophysiological importance in mental fatigue. Based on this literature and observations from our own laboratory and others on the role of astroglial cells in the fine-tuning of glutamate neurotransmission we present the hypothesis that the proinflammatory cytokines tumor necrosis factor-alpha, IL-1beta and IL-6 could be involved in the pathophysiology of mental fatigue through their ability to attenuate the astroglial clearance of extracellular glutamate, their disintegration of the blood brain barrier, and effects on astroglial metabolism and metabolic supply for the neurons, thereby attenuating glutamate transmission. To test whether our hypothesis is valid or not, brain imaging techniques should be applied with the ability to register, over time and with increasing cognitive loading, the extracellular concentrations of glutamate and potassium (K+) in humans suffering from mental fatigue. At present, this is not possible for technical reasons. Therefore, more knowledge of neuronal-glial signaling in in vitro systems and animal experiments is important.In summary, we provide a hypothetic explanation for a general neurobiological mechanism, at the cellular level, behind one of our most common symptoms during neuroinflammation and other long-term disorders of brain function. Understanding pathophysiological mechanisms of mental fatigue could result in better treatment. PMID- 15527506 TI - Allergic conditions and risk of hematological malignancies in adults: a cohort study. AB - BACKGROUND: Two contradictory hypotheses have been proposed to explain the relationship between allergic conditions and malignancies, the immune surveillance hypothesis and the antigenic stimulation hypothesis. The former advocates that allergic conditions may be protective against development of cancer, whereas the latter proposes an increased risk. This relationship has been studied in several case-control studies, but only in a few cohort studies. METHODS: The association between allergic conditions and risk of developing leukemia, Hodgkin's disease, non-Hodgkin's lymphoma and myeloma was investigated in a cohort of 16,539 Swedish twins born 1886-1925. Prospectively collected, self reported information about allergic conditions such as asthma, hay fever or eczema was obtained through questionnaires administered in 1967. The cohort was followed 1969-99 and cancer incidence was ascertained from the Swedish Cancer Registry. RESULTS: Hives and asthma tended to increase the risk of leukemia (relative risk [RR] = 2.1, 95% Confidence Interval [CI] 1.0-4.5 and RR = 1.6, 95% CI 0.8-3.5, respectively). There was also an indication of an increased risk of non-Hodgkin's lymphoma associated with eczema during childhood (RR = 2.3, 95% CI 1.0-5.3). CONCLUSION: In contrast to most previous studies, our results do not indicate a protective effect of allergic conditions on the risk of developing hematological malignancies. Rather, they suggest that allergic conditions might increase the risk of some hematological malignancies. PMID- 15527508 TI - Tongue lesions in psoriasis: a controlled study. AB - BACKGROUND: Our objective was to study tongue lesions and their significance in psoriatic patients. METHODS: The oral mucosa was examined in 200 psoriatic patients presenting to Razi Hospital in Tehran, Iran, and 200 matched controls. RESULTS: Fissured tongue (FT) and benign migratory glossitis (BMG) were the two most frequent findings. FT was seen more frequently in psoriatic patients (n = 66, 33%) than the control group (n = 19, 9.5%) [odds ratio (OR): 4.69; 95% confidence interval (CI): 2.61-8.52] (p-value < 0.0001). BMG, too, was significantly more frequent in psoriatic patients (28 cases, 14%) than the control group (12 cases, 6%) (OR: 2.55; 95% CI: 1.20-5.50) (p-value < 0.012). In 11 patients (5.5%), FT and BMG coexisted. FT was more frequent in pustular psoriasis (7 cases, 53.8%) than erythemato-squamous types (56 cases, 30.4%). On the other hand, the frequency of BMG increased with the severity of psoriasis in plaque-type psoriasis assessed by psoriasis area and severity index (PASI) score. CONCLUSIONS: Nonspecific tongue lesions are frequently observed in psoriasis. Further studies are recommended to substantiate the clinical significance of these seemingly nonspecific findings in suspected psoriatic cases. PMID- 15527507 TI - An enigmatic fourth runt domain gene in the fugu genome: ancestral gene loss versus accelerated evolution. AB - BACKGROUND: The runt domain transcription factors are key regulators of developmental processes in bilaterians, involved both in cell proliferation and differentiation, and their disruption usually leads to disease. Three runt domain genes have been described in each vertebrate genome (the RUNX gene family), but only one in other chordates. Therefore, the common ancestor of vertebrates has been thought to have had a single runt domain gene. RESULTS: Analysis of the genome draft of the fugu pufferfish (Takifugu rubripes) reveals the existence of a fourth runt domain gene, FrRUNT, in addition to the orthologs of human RUNX1, RUNX2 and RUNX3. The tiny FrRUNT packs six exons and two putative promoters in just 3 kb of genomic sequence. The first exon is located within an intron of FrSUPT3H, the ortholog of human SUPT3H, and the first exon of FrSUPT3H resides within the first intron of FrRUNT. The two gene structures are therefore "interlocked". In the human genome, SUPT3H is instead interlocked with RUNX2. FrRUNT has no detectable ortholog in the genomes of mammals, birds or amphibians. We consider alternative explanations for an apparent contradiction between the phylogenetic data and the comparison of the genomic neighborhoods of human and fugu runt domain genes. We hypothesize that an ancient RUNT locus was lost in the tetrapod lineage, together with FrFSTL6, a member of a novel family of follistatin-like genes. CONCLUSIONS: Our results suggest that the runt domain family may have started expanding in chordates much earlier than previously thought, and exemplify the importance of detailed analysis of whole-genome draft sequence to provide new insights into gene evolution. PMID- 15527509 TI - Computation of elementary modes: a unifying framework and the new binary approach. AB - BACKGROUND: Metabolic pathway analysis has been recognized as a central approach to the structural analysis of metabolic networks. The concept of elementary (flux) modes provides a rigorous formalism to describe and assess pathways and has proven to be valuable for many applications. However, computing elementary modes is a hard computational task. In recent years we assisted in a multiplication of algorithms dedicated to it. We require a summarizing point of view and a continued improvement of the current methods. RESULTS: We show that computing the set of elementary modes is equivalent to computing the set of extreme rays of a convex cone. This standard mathematical representation provides a unified framework that encompasses the most prominent algorithmic methods that compute elementary modes and allows a clear comparison between them. Taking lessons from this benchmark, we here introduce a new method, the binary approach, which computes the elementary modes as binary patterns of participating reactions from which the respective stoichiometric coefficients can be computed in a post processing step. We implemented the binary approach in FluxAnalyzer 5.1, a software that is free for academics. The binary approach decreases the memory demand up to 96% without loss of speed giving the most efficient method available for computing elementary modes to date. CONCLUSIONS: The equivalence between elementary modes and extreme ray computations offers opportunities for employing tools from polyhedral computation for metabolic pathway analysis. The new binary approach introduced herein was derived from this general theoretical framework and facilitates the computation of elementary modes in considerably larger networks. PMID- 15527510 TI - ESTIMA, a tool for EST management in a multi-project environment. AB - BACKGROUND: Single-pass, partial sequencing of complementary DNA (cDNA) libraries generates thousands of chromatograms that are processed into high quality expressed sequence tags (ESTs), and then assembled into contigs representative of putative genes. Usually, to be of value, ESTs and contigs must be associated with meaningful annotations, and made available to end-users. RESULTS: A web application, Expressed Sequence Tag Information Management and Annotation (ESTIMA), has been created to meet the EST annotation and data management requirements of multiple high-throughput EST sequencing projects. It is anchored on individual ESTs and organized around different properties of ESTs including chromatograms, base-calling quality scores, structure of assembled transcripts, and multiple sources of comparison to infer functional annotation, Gene Ontology associations, and cDNA library information. ESTIMA consists of a relational database schema and a set of interactive query interfaces. These are integrated with a suite of web-based tools that allow a user to query and retrieve information. Further, query results are interconnected among the various EST properties. ESTIMA has several unique features. Users may run their own EST processing pipeline, search against preferred reference genomes, and use any clustering and assembly algorithm. The ESTIMA database schema is very flexible and accepts output from any EST processing and assembly pipeline. ESTIMA has been used for the management of EST projects of many species, including honeybee (Apis mellifera), cattle (Bos taurus), songbird (Taeniopygia guttata), corn rootworm (Diabrotica vergifera), catfish (Ictalurus punctatus, Ictalurus furcatus), and apple (Malus x domestica). The entire resource may be downloaded and used as is, or readily adapted to fit the unique needs of other cDNA sequencing projects. CONCLUSIONS: The scripts used to create the ESTIMA interface are freely available to academic users in an archived format from http://titan.biotec.uiuc.edu/ESTIMA/. The entity-relationship (E-R) diagrams and the programs used to generate the Oracle database tables are also available. We have also provided detailed installation instructions and a tutorial at the same website. Presently the chromatograms, EST databases and their annotations have been made available for cattle and honeybee brain EST projects. Non-academic users need to contact the W.M. Keck Center for Functional and Comparative Genomics, University of Illinois at Urbana-Champaign, Urbana, IL, for licensing information. PMID- 15527511 TI - Elevated serum procollagen type III peptide in splanchnic and peripheral circulation of patients with inflammatory bowel disease submitted to surgery. AB - BACKGROUND: In the hypothesis that the increased collagen metabolism in the intestinal wall of patients affected by inflammatory bowel disease (IBD) is reflected in the systemic circulation, we aimed the study to evaluate serum level of procollagen III peptide (PIIIP) in peripheral and splanchnic circulation by a commercial radioimmunoassay of patients with different histories of disease. METHODS: Twenty-seven patients, 17 with Crohn and 10 with ulcerative colitis submitted to surgery were studied. Blood samples were obtained before surgery from a peripheral vein and during surgery from the mesenteric vein draining the affected intestinal segment. Fifteen healthy age and sex matched subjects were studied to determine normal range for peripheral PIIIP. RESULTS: In IBD patients peripheral PIIIP level was significantly higher if compared with controls (5.0 +/ 1.9 vs 2.7 +/- 0.7 microg/l; p = 0.0001); splanchnic PIIIP level was 5.5 +/- 2.6 microg/l showing a positive gradient between splanchnic and peripheral concentrations of PIIIP. No significant differences between groups nor correlations with patients' age and duration of disease were found. CONCLUSIONS: We provide evidence that the increased local collagen metabolism in active IBD is reflected also in the systemic circulation irrespective of the history of the disease, suggesting that PIIIP should be considered more appropiately as a marker of the activity phases of IBD. PMID- 15527514 TI - Adsorption of S-S containing proteins on a colloidal silver surface studied by surface-enhanced Raman spectroscopy. AB - We present a Raman and surface-enhanced Raman scattering (SERS) study of the following proteins containing S-S group(s): alpha chymotrypsin (alpha-CHT), insulin, lysozyme, oxytocin (OXT), Streptomyces subtilisin inhibitor (SSI), and trypsin inhibitor (STI). The SERS study is performed in order to understand the adsorption mechanism of the above-mentioned proteins on a colloidal silver surface. The SERS spectra presented here show bands associated mainly with aromatic amino acid vibrations. In addition, two distinct vibrations of the -C-S S-C- fragment are observed in the Raman and SERS spectra, i.e., nu(SS) and nu(CS). The enhancement of the nu(SS) vibration in the SERS spectra yields evidence that the intact disulfide bridge(s) is (are) located near the silver surface. This finding is supported by the presence of the nu(CS) mode(s). The presence of nus(COO-) and nu(C-COO-) in the SERS spectra in the 1384-1399 cm(-1) and 909-939 cm(-1) regions, respectively, indicate that the negatively charged COO- groups (aspartic and glutamic acids) assist in the binding on the positively charged silver surface. The Raman amide I and III bands observed in the 1621-1633 and 1261-1289 cm(-1) ranges, respectively, indicate that the alpha-helical conformation is favored for binding to the surface over the random coil or beta sheet conformations. In addition, the presence of the imino group of Trp and/or His indicates that these amino acid residues may also bind to the silver sol. PMID- 15527516 TI - Quantitative analysis of Raman signal enhancement from aqueous samples in liquid core optical fibers. AB - Raman scattering from aqueous liquids can be collected with high efficiency by enclosing the liquid within a suitable waveguide, as several groups have reported. Here, we present a quantitative model that predicts the relative strength of signals collected from (a) a tubular waveguide and (b) a flat-walled cuvette. Experimental measurements of Raman scattering from aqueous ethanol are made using two geometries, a Teflon-AF waveguide and a standard quartz cuvette. The model correctly predicts the enhancements in several ethanol Raman bands provided by the waveguide geometry. This model should be useful in aligning and characterizing liquid core waveguides, whose manufacture is still undergoing refinements. In particular, the model shows that absorption and scattering losses affect the enhancement factor in different ways. PMID- 15527515 TI - Factors affecting the sorption of model environmental pollutants onto silver polydimethylsiloxane nanocomposite Raman substrates. AB - The presence of aromatic compounds in water is an important topic in environmental sciences. Silver-polydimethylsiloxane nanocomposites (Ag-PDMS) have recently been demonstrated as promising substrates for the detection of model environmental pollutants via surface-enhanced Raman spectroscopy (SERS). This work discusses how different variables such as pH and matrix composition can affect the sorption and SERS activity of these chemicals. The results show that the conjugate base of weak acids can interact more efficiently with the substrate, leading to an increased signal at higher pH, while amino-aromatic compounds interact more efficiently at a lower pH. The sorption of these chemicals is an essential step in the process and has been attributed to the absorption of the analyte into the PDMS followed by its adsorption to the metallic surface. In addition, the presence of moderate concentrations (1 x 10( 4) M) of a supporting electrolyte such as nitrate or fluoride can improve the sorption of 4-hydroxybenzoic acid to the Ag-PDMS nanoparticles. Other ions such as phosphate and chloride cause rapid oxidation of the substrates even at concentrations as low as 1 x 10(-5) M. The effect of these variables in the analysis of real samples is presented. The potential use of liquid chromatography for isolating the model pollutants from detrimental matrix components in nat- ural waters is also shown. PMID- 15527517 TI - Analysis of passive mixing behavior in a poly(dimethylsiloxane) microfluidic channel using confocal fluorescence and Raman microscopy. AB - Confocal fluorescence microscopy (CFM) and confocal Raman microscopy (CRM) have been applied to monitor the laminar flow mixing behavior in a poly(dimethylsiloxane) (PDMS) microfluidic channel. Two passive PDMS micromixing devices were fabricated for this purpose: a two-dimensional round-wave channel and a three-dimensional serpentine channel. The microscale laminar flow mixing of ethanol and isopropanol was evaluated using the CFM and CRM at various flow rates. The mixing behavior of confluent streams in the microchannel was assessed by determining the degree of color change in Rhodamine 6G dye on mixing using the CFM. However, it was also possible to quantitatively evaluate the mixing process without employing a fluorescence label using the CRM. The results show a strong potential for CRM as a highly sensitive detection tool to measure fundamental fluid mixing processes and to provide detailed information on chemical changes of non-fluorescent reaction mixtures in a PDMS microfluidic channel. PMID- 15527518 TI - Application of fluorescence correlation spectroscopy for velocity imaging in microfluidic devices. AB - In this paper we present and demonstrate a technique for mapping fluid flow rates in microfluidic systems with sub-micrometer resolution using confocal microscopy in conjunction with fluorescence correlation spectroscopy (FCS). Flow velocities ranging from approximately 50 microm/s to approximately 10 cm/s can be recorded using fluorescent polymer nanospheres as fluid motion tracers. Velocity profiles and images of the flow in poly(dimethylsiloxane)-glass microchannels are presented and analyzed. Using the method, velocity images along the horizontal (top view) and vertical planes within a microdevice can be obtained. This is, to our knowledge, the first report of FCS for producing velocity maps. The high resolution velocity maps can be used to characterize and optimize microdevice performance and to validate simulation efforts. PMID- 15527520 TI - Molar absorptivities of glucose and other biological molecules in aqueous solutions over the first overtone and combination regions of the near-infrared spectrum. AB - Molar absorptivities are measured for water, glucose, alanine, ascorbate, lactate, triacetin, and urea in the near-infrared spectral region at 37 degrees C. Values are based on the Beer-Lambert law and cover the first overtone (1550 1850 nm; 6450-5400 cm(-1)) and combination (2000-2500 nm; 4000-5000 cm(-1)) spectral windows through aqueous media. Accurate calculations demand accounting for the impact of water displacement upon dissolution of solute. In this regard, water displacement coefficients are measured and reported for each solute. First overtone absorptivities range from 2 to 7 x 10(-5) mM(-1)mm(-1) for all solutes except urea, for which absorptivity values are below 0.5 x 10(-5) mM(-1) mm(-1) across this spectral range. Molar absorptivities over the combination spectral region range from 0.8 to 3.2 x 10(-4) mM(-1) mm(-1), which is a factor of four to five greater than the first overtone absorptivities. Accuracy of the measured values is assessed by comparing calculated or modeled spectra with spectra measured from standard solutions. This comparison reveals accurately modeled spectra in terms of magnitude and position of solute absorption bands. Both actual and modeled spectra from glucose solutions reveal positive and negative absorbance values depending on the measurement wavelength. It is shown that the net absorbance of light is controlled by the magnitude of the absorptivity of glucose compared to the product of the absorptivity of water and the water displacement coefficient for glucose. PMID- 15527521 TI - Quantifying ethanol content of beer using interpretive near-infrared spectroscopy. AB - On the basis of absorption measurements in the near-infrared (NIR) spectral range, a new method for the quantification of the ethanol content of beer is presented. Instead of the multivariate calibration models most commonly employed in NIR spectroscopic works, we use interpretive difference spectroscopy: Two wavelengths are selected according to the assignment of the absorption bands of the main substances of content of beer in the NIR region, and the difference between the absorbances at these wavelengths is used for ethanol quantification. Absorption spectra of the dominating beer ingredients are discussed and the calibration procedure with ethanol/water mixtures is shown. Robustness against the carbohydrate content of beer samples was demonstrated by analyzing solutions of ethanol and maltose in water. Validation of the method was performed with various beer samples with an ethanol concentration range between 0.5 and 7.7 vol %. The pertinent advantage of the procedure developed in this work is the indication that the results are independent from seasonal variations of the ingredients, which is of high interest for products with natural ingredients such as beer. PMID- 15527524 TI - Using the linearization approach for synchronizing the phase of photoacoustic reference and sample data. AB - Linearization of photoacoustic spectra is a technique that allows for detection of surface layers and the compensation of saturation effects in photoacoustic spectra by combining amplitude and phase information. The objective of this report is a thorough description of the linearization procedure. The influence of interferogram rotation on the real and imaginary parts of the complex Fourier transformation and on spectral phase angles is shown. The possibility of using the linearization approach to correct for phase differences between reference and sample data is discussed. The method can be used to calculate phase spectra from photoacoustic spectra taken with rapid scan spectrometers. PMID- 15527525 TI - Fourier transform infrared-probed O(3P) microreactor: demonstration with ethylene reactions in argon matrix. AB - To demonstrate the development of an oxygen atom microreactor in the form of liquid-helium-cooled solid argon matrix deposited on an infrared (IR) window, the oxidation of ethylene by mobile O atoms has been investigated. O atom diffusion through the argon matrix is confirmed and used to examine ethylene-oxygen atom reactions. In a bench-scale matrix isolation system probed with a Fourier transform infrared (FT-IR) spectrometer, matrices of solid Ar at 8-10 K doped with NO2 and ethylene have been prepared on a ZnSe window within an evacuated cryostat. The matrices have been photolyzed using 350-450 nm photons, and the reaction products resulting from the reaction of O(3P), one of the photolysis products of NO2, with ethylene have been identified using FT-IR and a Gaussian 98W simulation program. These products include oxirane, acetaldehyde, ethyl nitrite radical, and ketene. The temperature effect in the range of 10-30 K on the products formed has also been investigated. The reaction mechanisms are discussed and the viability of the solid Ar matrix being a low temperature microreactor to examine reaction mechanisms of mobile oxygen atoms is elaborated. PMID- 15527527 TI - Speciation of selenium in the mushroom Boletus edulis by high-performance liquid chromatography coupled to inductively coupled plasma-mass spectrometry with a collision cell. PMID- 15527529 TI - Nondestructive analysis of crystals within rat liver and duodenum samples by spectroscopic means. PMID- 15527530 TI - Personality and coping: a context for examining celebrity worship and mental health. AB - The adaptational-continuum model of personality and coping suggests a useful context for research areas that emphasize both personality and coping. The present paper used Ferguson's (2001) model integrating personality and coping factors to further conceptualize findings around celebrity worship. Three hundred and seventy-two respondents completed measures of celebrity worship, personality, coping style, general health, stress, positive and negative affect and life satisfaction. Celebrity worship for intense-personal reasons is associated with poorer mental heath and this relationship can be understood within the dimensions of neuroticism and a coping style that suggests disengagement. Such findings suggest the utility of examining the relationship between celebrity worship and mental health within both personality and coping variables, which have practical implications for understanding and addressing mental health problems that may occur as the result of engaging in celebrity worship for intense-personal reasons. PMID- 15527531 TI - Linguistic relativism and colour cognition. AB - Native speakers of two languages (English and Ndonga) were compared on three colour cognition tasks (sorting, triads and visual search) in a test of the linguistic relativity hypothesis (Whorf, 1956). The colour lexicons of these two languages differ because Ndonga has no basic terms for ORANGE, PINK and PURPLE, and stimuli were chosen to exploit this difference. On the sorting task (sorting into similarity-groups) for each language, nominally similar colours were grouped together more often than nominally dissimilar colours. On the triads task (choosing the most different of three colours), when the most nominally isolated colour differed for the two language-groups, each group tended to choose their nominal isolate. On the search task (scanning for target colours among distractors), targets were either in a different English category than distractors (cross-category), or some distractors were in the same English category as distractors (within-category). The 'cost' in speed of having within category distractors was much greater for the English than for the Ndonga. Overall, these data suggest that a core universal component is modulated by a small relativist influence. The differences in the visual search task are consistent with language affecting pre-attentive processes (an indirect language effect) as well as exerting on-line influences (a direct effect). PMID- 15527532 TI - Evidence for executive deficits among users of MDMA (Ecstasy). AB - Random letter generation and computation span are tasks known to load on executive, prefrontal resources. Previous research suggests that Ecstasy users are impaired on random letter generation. The current study, employing a larger sample (44 current Ecstasy users, and 59 non-Ecstasy users), together with more effective statistical controls for other drug use, failed to replicate previous findings. Ecstasy users were unimpaired on all measures of random generation performance. A significant difference was obtained on the computation span measure, with Ecstasy users scoring significantly lower than non-Ecstasy users. This difference remained statistically significant following control for various indicators of the use of other drugs including cannabis. The results are discussed in terms of the potential effects that Ecstasy might have on different component executive processes. PMID- 15527533 TI - Understanding the relationship between repetition priming and mere exposure. AB - Over the last two decades interest in implicit memory, most notably repetition priming, has grown considerably. During the same period, research has also focused on the mere exposure effect. Although the two areas have developed relatively independently, a number of studies has described the mere exposure effect as an example of implicit memory. Tacit in their comparisons is the assumption that the effect is more specifically a demonstration of repetition priming. Having noted that this assumption has attracted relatively little attention, this paper reviews current evidence and shows that it is by no means conclusive. Although some evidence is suggestive of a common underlying mechanism, even a modified repetition priming (perceptual fluency/attribution) framework cannot accommodate all of the differences between the two phenomena. Notwithstanding this, it seems likely that a version of this theoretical framework still offers the best hope of a comprehensive explanation for the mere exposure effect and its relationship to repetition priming. As such, the paper finishes by offering some initial guidance as to ways in which the perceptual fluency/attribution framework might be extended, as well as outlining important areas for future research. PMID- 15527534 TI - A model of aesthetic appreciation and aesthetic judgments. AB - Although aesthetic experiences are frequent in modern life, there is as of yet no scientifically comprehensive theory that explains what psychologically constitutes such experiences. These experiences are particularly interesting because of their hedonic properties and the possibility to provide self-rewarding cognitive operations. We shall explain why modern art's large number of individualized styles, innovativeness and conceptuality offer positive aesthetic experiences. Moreover, the challenge of art is mainly driven by a need for understanding. Cognitive challenges of both abstract art and other conceptual, complex and multidimensional stimuli require an extension of previous approaches to empirical aesthetics. We present an information-processing stage model of aesthetic processing. According to the model, aesthetic experiences involve five stages: perception, explicit classification, implicit classification, cognitive mastering and evaluation. The model differentiates between aesthetic emotion and aesthetic judgments as two types of output. PMID- 15527535 TI - The relation of depression and anxiety to life-stress and achievement in students. AB - OBJECTIVES: An apparent increase in seriously disturbed students consulting student health services in the UK has led to concern that increasing financial difficulties and other outside pressures may affect student mental health and academic performance. The current research investigated whether student anxiety and depression increases after college entry, the extent to which adverse life experiences contribute to any increases, and the impact of adversity, anxiety and depression on exam performance. METHOD: 351 UK-domiciled undergraduates completed questionnaires one month before university entry and mid-course. The Hospital Anxiety and Depression Scale (HADS: Zigmond & Snaith, 1983) was administered at both time points and a modified List of Threatening Experiences (Brugha, Bebbington, Tennant, & Hurry, 1985) was administered mid-course. RESULTS: By mid course 9% of previously symptom-free students became depressed and 20% became anxious at a clinically significant level. Of those previously anxious or depressed 36% had recovered. After adjusting for pre-entry symptoms, financial difficulties made a significant independent contribution to depression and relationship difficulties independently predicted anxiety. Depression and financial difficulties mid-course predicted a decrease in exam performance from first to second year. CONCLUSIONS: This is the first study to confirm empirically that financial and other difficulties can increase British students' levels of anxiety and depression and that financial difficulties and depression can affect academic performance. However, university life may also have a beneficial effect for some students with pre-existing conditions. With widening participation in higher education, the results have important implications for educational and health policies. PMID- 15527536 TI - The British Journal of Psychology centenary: a preliminary content survey and its problems. AB - The preliminary findings are reported of a review of the contents of the British Journal of Psychology 1904-2003 undertaken to mark the journal's centenary. This identifies (a) the top 11 categories of paper published and their patterns of distribution, (b) an apparent change in the nature of papers published after 1983, (c) the overall pattern of category-introduction, showing a dramatic change in 1965 and (d) a provisional typology of the principal patterns of distribution over time. Major conceptual and practical problems arising in the categorization of papers, and their implications, are discussed. Some reflections are offered on the possible value of research of this kind and on the nature of the journal itself. PMID- 15527537 TI - Using performance indicators to improve performance. AB - The recent introduction of performance assessment within the UK NHS, incorporating numerous performance indicators (PIs) and league tables, has led to health care organisations facing large numbers of targets and a star rating system with associated rewards and penalties. However, there is considerable evidence that using PIs for judgement rather than learning provides perverse incentives and can prove counterproductive. Drawing on earlier PI systems which, supplemented by expert systems, were designed to promote learning and exploration particularly by encouraging analysis of interactions between different indicators, a series of 'mini case studies' is presented. These reveal interesting relationships and suggest explanations for variations in performance, areas worth exploring further and possible approaches to improving performance -- approaches not apparent from individual indicators and league tables. It is concluded that presentation of PIs in a format that encourages exploration and analysis could greatly enhance the potential of the current PIs to improve NHS performance. PMID- 15527538 TI - The measurement and management of service quality in dental healthcare. AB - This paper reports the findings of a research study designed to manage and measure service quality in dental healthcare. Two large dental practices located in a major UK city were chosen for the research, and 300 patients from each of the two surgeries participated in the study. The findings provide insights into the appropriateness of the SERVQUAL framework (Parasuraman et al., 1988) as a tool for measuring service quality in the context of two distinct dental healthcare practices, one public and one private. The findings suggest that 'price' and 'experience' are two variables that may have an influence on service quality perceptions, when measured by customers. From the data, it was possible to identify service quality gaps where improvements could be made, and where resources could be re-allocated to obtain more optimum results in terms of enhancing service. In summary, the findings provide support for the SERVQUAL framework as a management tool for both measuring service quality and highlighting areas where improvements could be made within a dental healthcare environment. PMID- 15527539 TI - Do HMO penetration and hospital competition impact quality of hospital care? AB - This study examines the impact of HMO penetration and competition on hospital markets. A modified structure-conduct-performance paradigm was applied to the health care industry in order to investigate the impact of HMO penetration and competition on risk-adjusted hospital mortality rates (i.e. quality of hospital care). Secondary data for 1957 acute care hospitals in the USA from the 1991 American Hospital Association's Annual Survey of Hospitals were used. The outcome variables were risk-adjusted mortality rates in 1991. Predictor variables were market characteristics (i.e. managed care penetration and hospital competition). Control variables were environmental, patient, and institutional characteristics. Associations between predictor and outcome variables were investigated using statistical regression techniques. Hospital competition had a negative relationship with risk-adjusted mortality rates (a negative indicator of quality of care). HMO penetration, hospital competition, and an interaction effect of HMO penetration and competition were not found to have significant effects on risk adjusted mortality rates. These findings suggest that when faced with intense competition, hospitals may respond in ways associated with reducing their mortality rates. PMID- 15527540 TI - Factors influencing turnover and retention of midwives and consultants: a literature review. AB - This study is a review of literature on the factors affecting the retention and turnover of hospital consultants and midwives. While there is widespread concern and acknowledgement of staff retention problems for professional occupations within the NHS, far less research has analysed the causes of the staff retention problems for the occupations in question. This study shows that there is a dearth of literature in this area and that systematic comparative analysis of retention and turnover factors through both primary and secondary research is urgently required in order that policy-making can take place on the basis of informed choice. Tentative initial findings were that lack of appreciation or perceptions of not being valued are key factors influencing turnover for both occupations. Working hours, workload and work schedules are also common concerns to both groups. In addition, career development, promotion and appreciation of contribution were important retention factors for midwives, while a supportive professional environment, reduction in workload and working hours and more flexible work patterns were important to consultants. PMID- 15527541 TI - Analysing spatial trends in referral patterns to cancer genetics services: a preliminary investigation of regional variations in Wales. AB - This paper discusses spatial trends in referral patterns to a cancer genetics service. It presents a literature review outlining the paucity of existing research, a preliminary analysis at the Unitary Authority level in Wales and advances a programme of further research to be conducted at a more detailed spatial level. The preliminary analysis shows a weak negative relationship between referral rates from primary care and social deprivation by Unitary Authority (Spearman rank correlation coefficient, sigma = -0.38). There is also a weak positive relationship between average settlement size and referral rates (sigma = +0.28), which taken together may indicate that primary care practices in affluent urban areas are more likely to refer than those in poorer rural areas. Future research will be conducted at a finer spatial scale, and will take into account characteristics of primary care practices and the patients being referred, amongst other variables. PMID- 15527542 TI - Realities of respite care. AB - Respite care in the form of day care centres, in-home respite and institutional care, has been the cornerstone of care for the disabled elderly. Evidence for its effectiveness in helping carers or disabled clients is hard to come by. In this paper, a proposition is put forward for a carefully designed study to answer some of the important points often raised by the providers of such services. PMID- 15527543 TI - New combinations of anti-inflammatories for therapy of arthritic, neurological and malignant diseases. PMID- 15527545 TI - Plasma proteins as indices of physiologic age and cumulative stress, as well as of disease, inflammation, infection and injury. PMID- 15527546 TI - Anti-TNF-alpha therapy for chronic inflammation: reconsidering pentoxifylline as an alternative to therapeutic protein drugs. AB - Pentoxifylline is a useful inhibitor of TNF-alpha production, thus resembling anti-inflammatory corticosteroids. The combination of pentoxifylline with Lyprinol is an effective treatment for chronic inflammation in rats, in this respect mimicking low dose prednisone with Lyprinol. PMID- 15527547 TI - ANA+ (anti-nuclear antibody) is correlated with RF+ (rheumatoid factor) in rheumatoid arthritis. PMID- 15527548 TI - Src-kinase-dependent epidermal growth factor receptor transactivation in salivary mucin secretion in response to beta-adrenergic G-protein-coupled receptor activation. AB - The principal regulatory factors that control the flow and make-up of salivary secretion are neurotransmitters, released by parasympathetic and sympathetic innervation, that trigger activation of G protein-coupled receptors (GPCRs) on the acinar cells of salivary glands and stimulate the generation of soluble second messengers. In this study, we report that activation of GPCR by beta adrenergic agonist leading to stimulation in salivary mucin secretion occurs with the involvement of epidermal growth factor receptor (EGFR). Using [(3)H]glucosamine-labeled mucous acinar cells of sublingual salivary gland in culture, we show that stimulatory effect of beta-adrenergic agonist, isoproterenol, on mucin secretion was inhibited in a concentration-dependent manner by EGFR kinase inhibitor, PD153035, as well as wortmannin, an inhibitor of PI3K. Moreover, both inhibitors caused the impedance in the acinar cell mucin secretory responses to beta-adrenergic agonist-generated second messenger, cAMP, as well as adenylate cyclase activator, forskolin. The acinar cell secretory responses to isoproterenol, furthermore, were blunted in a concentration dependent fashion by PP2, a selective inhibitor of tyrosine kinase Src responsible for ligand-independent EGFR autophosphorylation. However, no significant alterations in the acinar cell mucin secretory responses to isoproterenol, cAMP or forskolin were attained with an inhibitor of the ERK pathway, PD98059. Our findings underline the role of EGFR as a convergence point in modulation of salivary mucin secretion triggered by beta-adrenergic agonist GPCR activation and demonstrate the importance of Src kinase in the EGFR transactivation process. PMID- 15527549 TI - Effect of Mycobacterium leprae lipids on BCG- and carrageenan-induced cellular recruitment in mouse pleurisy. AB - Pathogenic mycobacteria survive inside macrophages and deactivate these cells, using a mechanism that is still poorly understood. Mycobacterial cell wall lipids constitute the first contact with the host cell. Although Mycobaterium leprae and M. bovis BCG share common antigens, they induce opposite inflammatory responses. Apolar M. leprae lipids have been shown to be anti-inflammatory by down regulating macrophage activation and T-cell functions. We wonder if these lipids would influence cellular migration to BCG or to other inflammatory agent. We investigated the effect of M. leprae, its lipids or delipidated bacteria on acute and chronic BCG- or carrageenan-induced pleurisy. Previous injection of intact or delipidated M. leprae did not alter either the BCG- or carrageenan-induced pleural inflammatory reaction. However, M. leprae lipids enhanced carrageenan induced acute cellular migration without impairing BCG inflow; moreover, they reduced BCG chronic response. Together these data suggest distinct mechanisms for intracellular deactivation and pleural cell recruitment exerted by mycobacterial structures. PMID- 15527550 TI - Cicatrizing and antimicrobial properties of an ozonised oil from sunflower seeds. AB - The ozonised sunflower oil, Bioperoxoil, was tested for its antimicrobial activity against some pathological strains in vitro together with its healing potential against Staphylococcus aureus in vivo. Bioperoxoil was tested against S. aureus, Pseudomonas aeroginosa, Candida albicans, S. typhimurium and Escherichia coli suspensions using the agar diffusion method. Healing experiments were carried out with Wistar rats through topical application of 3.5 mg/ml of the ozonised oil up to the 7th day after inoculation with S. aureus. Bioperoxoil showed anti-inflammatory effects against all strains tested, with MIC values ranging from 2.0 to 3.5 mg/ml. Bioperoxoil also demonstrated protective effects on skin connective tissue and to enhance wound healing during the treatment, as compared to a neomycin-clostebol association used as a positive control. The overall results indicated a significant antimicrobial activity, anti-inflammatory and wound-healing properties for Bioperoxoil, as compared to other antimicrobial agents commercially available. PMID- 15527551 TI - Anti-inflammatory therapy with a COX-2 inhibitor in Tourette's syndrome. AB - An infectious/inflammatory process plays a role in at least a subgroup of patients with tics and Tourette's syndrome (TS). Successful antibiotic therapy and prophylaxis was described repeatedly. We report the case of a patient suffering from chronic TS who was treated with celecoxib additionally to the antibiotic prophylaxis. This treatment was associated with a continuous improvement of tics and disturbed behaviour, such as aggression and social withdrawal. The withdrawal of celecoxib led to a marked deterioration in TS symptoms while the re-exposition had advantageous therapeutic effects. This result of the treatment with a COX-2 inhibitor supports the view that COX-2 inhibitors show therapeutic benefit in patients suffering from psychiatric disorders in which an inflammatory process is involved in the pathophysiology. PMID- 15527552 TI - Role of histamine and acid back-diffusion in modulation of gastric microvascular permeability and haemorrhagic ulcers in betel-quid-fed rats. AB - Evidence concerning the pathogenesis of gastric haemorrhagic ulcer produced by betel quid chewing (BQC) is lacking. This research first proposes that alterations of mast cell histamine release and gastric acid back-diffusion are important in modulating gastric microvascular permeability and mucosal haemorrhagic ulcer in BQC-fed rats. The effects of several histamine receptor antagonists on this ulcer model also were evaluated. Male Wistar rats were fed with BQC diet or normal pellet diet. After 1, 30 and 90 day(s), rat stomachs were irrigated for 3 h with either normal saline or simulated gastric juice. Gastric acid back-diffusion, mucosal histamine concentration, microvascular permeability, as well as luminal haemoglobin content and ulcer areas were determined. Severe gastric haemorrhage and mucosal ulcerations, particularly in acidic stomachs, were observed in BQC-fed rats. A high correlation was observed between histamine and gastric haemorrhage, as well as between acid back-diffusion and mucosal ulceration was found in rats fed with BQC. This haemorrhagic ulcer in BQC-fed rats was effectively ameliorated by intragastric ketotifen, ranitidine or their combination. In conclusion, enhancement of acid back-diffusion, mast cell histamine release and microvascular permeability is important in modulating gastric haemorrhage and ulcer in BQC-fed rats. PMID- 15527573 TI - A head and neck cancer database for the 21st century. AB - A versatile, hand-held computerised database is described. Its use for general patient management, audit and as a tool for clinical governance is illustrated. PMID- 15527574 TI - The information needs of head and neck cancer patients prior to surgery. AB - OBJECTIVE: To describe the common themes in the experiences and expressed information needs of patients undergoing head and neck surgery. Summary background data : Patients who suffer head and neck cancers and undergo surgery often report considerable psychological distress and impaired social functioning. To optimise survival, the decision about what treatment option to follow is often made quickly, with little support in terms of counselling or the provision of information. There is inadequate previous work exploring the content and delivery of information required by patients at this time. PATIENTS AND METHODS: Participants included patients who had undergone surgery for head or neck cancer (n=29) and their immediate relatives who were present at the initial consultation with the surgeon (n=13). Patients were recruited from out-patient departments in two hospitals in the north of England. All interviews were conducted in participants' homes and were guided by a semistructured interview schedule devised both from literature and a pilot study. RESULTS: Whilst most participants felt well informed about the surgical procedure they were undergoing, many reported feeling unprepared for the long-term lifestyle changes that occurred. Information, support and advice throughout the 3-6 months postoperative period was reported to be inadequate. The majority of participants did not ask any questions and did not perceive there was a choice regarding treatment. Individuals who wanted to take an active role in decision-making reported difficulties accessing information to enable them to do so. CONCLUSION: The findings of this study emphasise the need for individualised information provision defined not exclusively by the surgical procedure. PMID- 15527575 TI - The surgical arrest of post-tonsillectomy haemorrhage: hospital episode statistics. AB - INTRODUCTION: In the consent of patients for tonsillectomy, the risk of returning to theatre for control of postoperative bleeding may be determined from data provided on Hospital Episode Statistics (HES) provided by the Department of Health website. METHODS: HES data for England from 1998-2002 were used. The numbers of tonsillectomies and of surgical arrest of post-tonsillectomy haemorrhage were considered for children and adults. RESULTS: Of 220,497 tonsillectomies performed over the 4-year period, 1804 (0.82%) returned to theatre for control of bleeding. However, this was 3.87 times more likely in adults than children (P <0.0001). CONCLUSION: Adults are nearly four times more likely than children to require surgery for the arrest of post-tonsillectomy haemorrhage. PMID- 15527576 TI - A survey of current myocardial protection practices during coronary artery bypass grafting. AB - OBJECTIVE: To identify current myocardial protection strategies for coronary artery bypass grafting (CABG) across the UK and Ireland. METHODS: A questionnaire survey of 15 questions was sent to practising cardiac surgeons between June and October 2002. The list of surgeons was obtained from the Society of Cardiothoracic Surgeons of Great Britain and Ireland database and they were contacted by postal and electronic mail. RESULTS: 118 (73.7%) out of 160 surgeons responded to the survey. 61 (51.7%) perform CABG on-pump (ONCAB) while 10 (8.5%) practice off-pump CABG (OPCAB). 47 (39.8%) perform either depending on individual cases. Of the 108 surgeons performing ONCAB, 91 (84.3%) use cardioplegia while 17 (15.7%) use cross-clamp and fibrillation techniques. Of those using cardioplegia, 76 (83.5%) use blood cardioplegia, 15 (19.7%) use warm-blood and 60 (78.9%) use cold-blood cardioplegia. 15(16.5%) use crystalloid cardioplegia. Retrograde cardioplegia is used by 23 (25.2%). We find an interesting variation of practice in relation to specifics like warm induction, graft cardioplegia, hot-shot, single cross-clamp, hypothermia and venting procedures. An overwhelming majority of surgeons performing OPCAB use the Octopus stabiliser (n=44, 77.2%) with some others preferring the Genzyme system. Supplementary stabilisation is not commonly used. While most OPCAB surgeons use intracoronary shunts (n=51), some prefer blockers (n=9) and others use coronary sloops (n=36). Ischaemic preconditioning is not commonly practised. Several surgeons have changed their practice of myocardial protection in the last 5 years (n=45). CONCLUSIONS: This survey gives us an interesting insight into current myocardial protection practices in the UK and Ireland and may be useful for future reference. PMID- 15527577 TI - Tuberculosis of the breast: experience of a UK breast clinic serving an ethnically diverse population. AB - Tuberculosis (TB) of the breast is an uncommon disease in the West but its incidence is likely to increase. Five cases of breast tuberculosis are presented. The diagnosis and management of this condition are discussed. PMID- 15527578 TI - Hepatic resection for colorectal metastases - a national perspective. AB - BACKGROUND: Many consultant surgeons are uncertain about peri-operative assessment and postoperative follow-up of patients for colorectal liver metastases, and indications for referral for hepatic resection. The aim of this study was to assess the views the consultant surgeons who manage these patients. METHODS: A postal questionnaire was sent to all consultant members of the Association of Coloproctology of Great Britain and Ireland and of the Association of Upper Gastrointestinal Surgeons of Great Britain and Ireland. The questionnaire assessed current practice for preoperative assessment and follow-up of patients with colorectal malignancy and timing of and criteria for hepatic resection of metastases. Number of referrals/resections were also assessed. RESULTS: The response rate was 47%. Half of the consultants held joint clinics with an oncologist and 89% assessed the liver for secondaries prior to colorectal resection. Ultrasound was used by 75%. Whilst 99% would consider referring a patient with a solitary liver metastasis for resection, only 62% would consider resection for more than 3 unilobar metastases. The majority (83%) thought resections should be performed within the 6 months following colorectal resection. During follow-up, 52% requested blood CEA levels and 72% liver ultrasound. Half would consider chemotherapy prior to liver resection and 76% performed at least one hepatic resection per year with a median number of 2 resections each year. CONCLUSIONS: A substantial proportion of patients are assessed for colorectal liver metastases preoperatively and during follow-up though there is spectrum of frequency of assessment and modality for imaging. Virtually all patients with solitary hepatic metastases are considered for liver resection. Patients with more than one metastasis are likely to be not considered for resection. Many surgeons are carrying out less than 3 resections each year. PMID- 15527579 TI - 'One-stop' inguinal hernia surgery--day-case referral, diagnosis and treatment. AB - BACKGROUND: Patients with unilateral inguinal hernias are conventionally seen at an out-patient appointment before being placed on a waiting list for surgery. Many patients are also required to attend a pre-assessment clinic prior to admission. AIM: To establish whether patients fit for day surgery inguinal hernia repair could be assessed and treated at a single hospital appointment. PATIENTS AND METHODS: Patients referred with unilateral inguinal hernias were sent an information sheet and health questionnaire by post. General practitioners were asked to help patients complete the medical details. Patients suitable for day surgery were sent a single appointment for assessment and treatment by an open, tension-free Lichtenstein mesh repair. RESULTS: Ninety-eight patients were sent an appointment for 'one-stop' inguinal hernia treatment. Ninety-two patients (93.5%) underwent inguinal hernia repair and were discharged on the same day without complication. CONCLUSIONS: Patients with unilateral primary inguinal hernias who are under 70 years old and ASA grade I or II can been seen, assessed and treated on the same day. 'One-stop' inguinal hernia surgery reduces the number of patient visits to the hospital and could be expanded to incorporate many more hernia repairs and other day case procedures. PMID- 15527580 TI - The changing practice of transurethral prostatectomy: a comparison of cases performed in 1990 and 2000. AB - OBJECTIVES: Transurethral resection of the prostate (TURP) is considered by many to be the 'gold standard' treatment for benign prostatic enlargement. However, with the relatively recent introduction of pharmacological and other surgical treatment modalities, the performance of TURP appears to be in decline. METHODS: A retrospective casenote analysis of 200 patients who underwent TURP in 1990 and the year 2000 with the aim of identifying changes in the incidence and practice of TURP. RESULTS: There was a decline in the number of TURPs performed of 31.6% over the 10-year period, with more being carried out because of urinary retention. In 2000, the patient was older and the operative procedure took statistically longer than 10-years earlier, but the weight of prostate tissue resected, patient satisfaction and complication rates were similar. CONCLUSIONS: At present, TURP is in decline, with urinary retention being the commonest indication. The population at present is older but this does not carry additional co-morbidity. The weight of resection has not altered, although surgery currently takes longer to perform. PMID- 15527581 TI - What is important to patients receiving extracorporeal shock-wave lithotripsy for urinary tract calculi? AB - INTRODUCTION: Patients' experience and expectations are paramount in the current era of clinical governance. However, there is lack of published information on patients' expectations from extracorporeal shock-wave lithotripsy (ESWL) treatment for renal stone disease. We conducted a survey to quantify what aspects of ESWL are important purely from the patients' prospective. PATIENTS AND METHODS: Postal questionnaires were sent out to all patients who attended our unit for ESWL treatment for renal stone disease. The questionnaire referred to 15 aspects of ESWL treatment. Patients were asked to rate each aspect based on a numerical scale from 1 to 10 (1--least important; 10--most important). RESULTS: We posted 340 questionnaires and received 208 valid replies (61%). Stone clearance was statistically the most important aspect of treatment. Staff attitude and explanation of the procedure and outcomes were rated highly. To our surprise, pain control was rated only 8th. CONCLUSIONS: Patients' priorities may be different to ours. Surveys like this allow ones to put the appropriate 'weight' on the results of patient satisfaction surveys in order to allocate resources and improve service appropriately. PMID- 15527582 TI - Metastatic bone disease--the orthopaedic workload. AB - BACKGROUND: In 1999, the British Association of Surgical Oncology (BASO) published its guidelines for the management of metastatic bone disease in breast cancer in the UK, highlighting the role of the orthopaedic surgeon as an integral part of the multidisciplinary team. AIM: To establish the workload to our department of metastatic bone disease. METHODS: A prospective study was performed over a 6-month period aiming to study factors affecting length of stay, whether BASO recommendations were being adhered to for metastatic breast disease and whether BASO recommendations could be extended to other forms of metastatic bone disease. RESULTS: 36 patients were referred to the orthopaedic department during the study period. The majority of the patients were referred as emergencies, 83% of patients were seen by an orthopaedic consultant on the day of referral but only 27% were seen by the orthopaedic cancer liaison consultant. Only 16 patients required a therapeutic operative intervention and length of in-patient stay was increased by not having a known primary carcinoma at admission. Patients with bone metastases discussed at the breast cancer multidisciplinary meeting were also studied as were patients with bone metastases presenting to the oncology service. It was found that there was little overlap between these groups and our study group. CONCLUSIONS: Metastatic bone disease does form a workload to the orthopaedic department and BASO recommendations are generally followed for breast disease. The BASO recommendations could be extended to other tumours; however, it should be recognised that there is a significant workload involved and a need for sufficient resources to allow a high quality and timely service. PMID- 15527583 TI - Total hip arthroplasty: results and consequences. PMID- 15527584 TI - Avulsion of the common origin of the medial collateral and medial patello-femoral ligaments. AB - This study documents for the first time avulsion of the common origin of the medial collateral and medial patello-femoral ligaments. PMID- 15527585 TI - Repair of acute rupture of the Achilles tendon: a new technique using polyester tape without external splintage. AB - INTRODUCTION: A new method of treating acute rupture of the Achilles tendon using polyester tape is reported. This requires no postoperative splintage, allows earlier mobilisation and prompt return to work and sport. PATIENTS AND METHODS: Thirty patients were reviewed prospectively and at a mean of 3 years from surgery. RESULTS: The average time of return to work, full weight-bearing and to driving was 40 days, 45 days and 49 days, respectively. The average time for return to sport was 122 days. Three patients required further surgery, two for infected wounds and one for scar release. There was one sural nerve injury. Twenty-two patients regained a normal range of ankle and subtalar movement, with the mean power of plantar flexion 84% of the opposite side. Of the 22 patients who played sport, 14 were still performing at the same or a higher level. There were no re-ruptures over this period. CONCLUSIONS: The technique is straightforward and avoids splintage. It also conveys other short- and long-term advantages over more established methods. PMID- 15527586 TI - Wound infections in day-case hand surgery: a prospective study. AB - BACKGROUND: There is little published information about postoperative infection rates in day-case hand surgery and the possible factors promoting infection. RESULTS: We report on infection rates observed in day-case hand surgery of 1035 cases over a 2-year period performed by two consultant surgeons. Infection rates of just over 1% were observed. CONCLUSION: We believe that this low infection rate reflects seniority of surgeon, operative technique and short duration of operation. PMID- 15527587 TI - Regional orthopaedic courses--rationale and practice. AB - BACKGROUND: The intensity and length of higher surgical training has changed, and with this the need for a more formalised training programme has been recognised. With limited budgets for training and the high cost of national training days, it was proposed that within the Wessex Deanery regional courses should be run to complement the existing weekly training session. METHODS: In 1998, a questionnaire was sent to all the orthopaedic consultants and specialist registrars within the region to identify the training requirements of the specialist registrars and the availability of consultants to organise and teach during these training courses. RESULTS: A regionally published report based on the questionnaire's findings identified these training requirements as indicated by both the specialist registrars and the consultants. This information has been used to organise eight 1-day training courses using the expertise of consultants within the region with more planned in the near future. CONCLUSIONS: Four years following this report, we identified the training requirements within the Wessex region and have successfully run regional courses to meet this requirement. PMID- 15527588 TI - Is there a need for resident general surgical cover in small peripheral hospitals? AB - AIM: To assess the need for resident general surgical cover in a small peripheral hospital. PATIENTS AND METHODS: The total number of admissions to Caerphilly District Miners' Hospital in the year 2001 was noted along with the admission criterion for elective general and vascular surgical patients. RESULTS: Among the 10,608 in-patients only 120 (1.13%) developed general surgical/vascular problems that merited surgical referral and out of these 30 (0.28%) patients were transferred to neighbouring larger hospitals for specialist care. CONCLUSIONS: A resident staff grade surgeon is not required in a small peripheral hospital and this service could be provided by the resident on-call surgical SpR in a neighbouring larger hospital. PMID- 15527589 TI - New consent forms issued by the Department of Health. PMID- 15527590 TI - Improving risk disclosure during the consent process. AB - OBJECTIVES: To provide guidance about the risks which should be disclosed to patients and documented during the consent process. METHODS: The Delphi Consensus Technique was used to decide what constitutes mandatory risk disclosure for three index procedures. Documentation of risk on consent forms was audited and compared to these locally agreed standards. A four stage strategy for change was undertaken following which practice was reviewed. RESULTS: Mean mandatory risk documentation rose from 61.2% (95% CI: 58.1-64.4) pre-intervention, to 78.1% (95% CI: 72.6-83.6) post-intervention (ccc2; P < 0.001). CONCLUSIONS: Although we demonstrated some benefit from this simple approach, the need for pragmatic means of achieving and sustaining complete discussion and documentation of risks across all surgical interventions based on universally accepted standards remains. PMID- 15527591 TI - 'Will you be doing my operation doctor?' Patient attitudes to informed consent. AB - BACKGROUND: As part of the consent process, it is part of a doctor's duty of care to reveal any material risk. Depending upon the level of supervision, whether the operating surgeon is a trainee may be such a risk, but in our experience this is not routinely discussed with patients pre-operatively. We set out to discover patients' attitudes to being operated on by trainee urological surgeons. PATIENTS AND METHODS: A total of 101 completed questionnaires were received from patients (90 male, 11 female, mean age 72 years) undergoing transurethral resection of the prostate (TURP), transurethral resection of a bladder tumour (TURBT) or cystodiathermy on various aspects of their attitudes to being operated on by junior doctors as part of training. RESULTS: The response rate was 77%. Of the respondents, 94 patients (91%) thought that junior doctors should perform surgery as part of their training. Only 11 of 73 (15%) said they would be happy for a junior doctor, competent to perform the procedure, to operate unsupervised. Of 98 patients, 80 (82%) thought they should be told if the operation was going to be performed by a junior doctor, and 85 (87%) that they should be told their name and designation. CONCLUSIONS: For consent to be 'informed', the experience and identity of the surgeon should be made known to patients. Most patients are happy to be operated on by a junior doctor under consultant supervision, but would want to be told and know their name and status. PMID- 15527592 TI - Response to paper by T Ibrahim, SM Ong, GJStC Taylor the new consent form: is it any better? Ann R Coll Surg Engl 2004; 86: 206-9: response 1. PMID- 15527593 TI - Informed consent and surgeons in training: do patients consent to allow surgical trainees to operate on them? PMID- 15527594 TI - Response to paper by T Ibrahim, SM Ong, GJStC Taylor the new consent form: is it any better? Ann R Coll Surg Engl 2004; 86: 206-9: response 3. PMID- 15527595 TI - Response to paper by T Ibrahim, SM Ong, GJStC Taylor the new consent form: is it any better? Ann R Coll Surg Engl 2004; 86: 206-9: response 2. PMID- 15527597 TI - Common bile duct calculi--ERCP vs laparoscopic exploration: the case for endoscopic retrograde cholangiopancreatography (ERCP). PMID- 15527601 TI - Hand-held Doppler to assess valvular incompetence--a new OSCE station? PMID- 15527602 TI - Response to paper by SP Smith Olecranon fractures--a reliable technique for accurate positioning of the tension band wire. PMID- 15527604 TI - Should we stop telling well pregnant women to monitor fetal movements? How to use and interpret guidelines. PMID- 15527605 TI - Republic of Ireland's indoor workplace smoking ban. PMID- 15527606 TI - Positive experiences of teenage motherhood: a qualitative study. AB - BACKGROUND: Teenage pregnancy is seen as a cause for concern in the United Kingdom (UK). However, there has been little research from primary care looking at teenage motherhood and its implications. AIM: To investigate the experiences of teenage mothers in relation to their role as mothers and their expectations of their futures. DESIGN OF STUDY: Qualitative study. SETTING: East Devon, England. METHOD: Nine women who had conceived their first child while still a teenager agreed to participate. Semi-structured interviews were undertaken, audiotaped, transcribed, and analysed using interpretative phenomenological analysis. RESULTS: The women expressed positive attitudes to being mothers and described how it had affected their lives. For some, motherhood had been the impetus to change direction and consider a career, because they had someone else for whom they were responsible. They recognised that they were still young enough to enter further education or other aspects of employment as their children grew up. CONCLUSIONS: For the women in this study, having been a teenage mother did not mean that their life and future were all over. Motherhood and bringing up children were valued in their own right. The women were realistic about their futures, often making plans to develop their careers. PMID- 15527607 TI - Randomised controlled trial of pelvic floor muscle exercises and manometric biofeedback for erectile dysfunction. AB - BACKGROUND: The pelvic floor muscles are active in normal erectile function. Therefore, it was hypothesised that weak pelvic floor muscles could be a cause of erectile dysfunction. AIMS: To compare the efficacy of pelvic floor muscle exercises and manometric biofeedback with lifestyle changes for men with erectile dysfunction. DESIGN OF STUDY: Randomised controlled trial. SETTING: The Somerset Nuffield Hospital, Taunton, United Kingdom. METHOD: Fifty-five men with erectile dysfunction (median age 59.2 years; range 22-78 years) were enrolled from a local urology clinic. Of these, 28 participants were randomised to an intervention group and engaged in pelvic floor exercises, as well as receiving biofeedback and suggestions for lifestyle changes. Twenty-seven controls were solely advised on lifestyle changes. Baseline, 3- and 6-month assessments were: erectile function domain of International Index of Erectile Function (IIEF), Partner's International Index of Erectile Function (PIIEF), Erectile Dysfunction-Effect on Quality of Life (ED-EQoL), anal manometry, digital anal measurements, and clinical assessment by an assessor blind to treatment allocation. After 3 months, the control group were transferred to the active arm. RESULTS: At 3 months, compared with controls, men in the intervention group showed significant mean increases in the erectile function domain of the IIEF (6.74 points, P = 0.004); anal pressure (44.16 cmH(2)O, P <0.001); and digital anal grades (1.5 grades, P <0.001). All showed further improvement in these outcomes at 6 months. Similar benefits were seen in men of the control arm after transfer to active treatment. A total of 22 (40.0%) participants attained normal function, 19 (34.5%) participants had improved erectile function, and 14 (25.5%) participants failed to improve. CONCLUSION: Pelvic floor muscle exercises and biofeedback are an effective treatment for men with erectile dysfunction. PMID- 15527608 TI - The health-related quality of life of people with peripheral arterial disease in the community: the Edinburgh Artery Study. AB - BACKGROUND: Previous studies investigating the health-related quality of life of those with peripheral arterial disease have focused on patients recruited from hospital clinics. The health-related quality of life of people with peripheral arterial disease in the general population is unknown. AIMS: We aimed to determine the health-related quality of life of people with intermittent claudication and asymptomatic peripheral arterial disease in the general population and to compare it with those with angina and those with no peripheral arterial disease or angina. DESIGN OF STUDY: Analysis of cross-sectional data from the 12-year follow-up of a population-based cohort. SETTING: Edinburgh, Scotland. METHOD: Data from the Edinburgh Artery Study cohort's 12-year follow-up was analysed. Participants' peripheral arterial disease status was measured using the World Health Organisation intermittent claudication questionnaire and the ankle brachial pressure index. Self-assessed health-related quality of life data was collected using the SF-36 generic questionnaire. Health-related quality of life scores were calculated and their associations with peripheral arterial disease status groups were tested. RESULTS: Subjects with intermittent claudication had significantly worse median health-related quality of life scores than patients without claudication in all domains except social functioning and mental health. Patients with claudication had a significantly lower physical component summary score than those without claudication (P /= 16 years scoring >/= 12 on the Clinical Interview Schedule Revised were randomised. The experimental intervention required participants to complete a computerised psychosocial assessment that generated a report for the GP including patient-specific treatment recommendations. The control patients were treated as usual with access to locally agreed guidelines. RESULTS: Participants' 12-item General Health Questionnaire (GHQ) score dropped irrespective of treatment allocation. The experimental group had a significantly lower GHQ score at 6 weeks, but not at 6 months. Recovery at 6 months was 3% greater among those receiving the experimental intervention (95% confidence interval [CI] = -4 to 10). Treatment was not significantly associated with quality of life or patient satisfaction. CONCLUSION: Only small benefits are likely from using case-finding followed by patient-specific guidelines to improve clinical management of common mental disorders in primary care. However, depression and anxiety are important public health problems so the utility of such systems should be further investigated. PMID- 15527610 TI - General practitioners' knowledge of post-traumatic stress disorder: a controlled study. AB - BACKGROUND: Post-traumatic stress disorder (PTSD) is common, is associated with substantial morbidity, and is often not recognised in primary care. AIM: To explore whether general practitioners (GPs) have significant gaps in their knowledge of PTSD. DESIGN OF STUDY: A controlled study. SETTING: Primary care in two Scottish regions. METHOD: A validated postal questionnaire consisting of clinical vignettes for PTSD, acute stress reaction, and depression was used to gather the data. The primary outcome measures were the proportion describing 'best practice' management of PTSD and the comparison of this with the control condition, the proportion describing 'best practice' management of depression. The secondary outcome measures were comparisons of PTSD and depression by recognition, drug treatment, and referral. RESULTS: Two-thirds (67.5%) of GPs included PTSD in their differential diagnosis for the PTSD vignette, and 86.8% made a referral to secondary care for the PTSD case. A minority of GPs (42.9%) and only 54.1% of a comparison group of psychiatrists specified the drug treatment of choice for PTSD, a selective serotonin reuptake inhibitor. Only 28.3% of GPs had the knowledge to recognise PTSD and prescribe appropriately, compared with 89.8% for depression (P <0.001). Only 10.2% of GPs described best practice for PTSD, compared with 47.7% for depression (P <0.001). CONCLUSION: Lack of knowledge is among the reasons for less than ideal recognition and management of PTSD in primary care. Further research should aim to explore the implementation of PTSD guidelines in primary care. PMID- 15527611 TI - Dutch general practitioners' referral of children to specialists: a comparison between 1987 and 2001. AB - BACKGROUND: Although children are frequently referred to specialists, detailed information on referral patterns of them is scarce. Even less information is available on how referral patterns evolve over time. AIMS: To examine current referral patterns for children aged 0-17 years and compare these with referral patterns reported for 1987. DESIGN OF STUDY: Data were analysed from two national cross-sectional surveys, performed in 2001 (91 general practices) and in 1987 (103 general practices). SETTING: Dutch general practice. METHOD: All new referrals to specialists were assessed by age, sex, International Classification of Primary Care (ICPC) category, specialty referred to, and specific episodes of disease. Referral measures were quantified as new referrals per 1000 person-years and per 100 new episodes, a measure of likelihood of a young person with a specific diagnosis to be referred. Rates in 2001 were compared with those from 1987. RESULTS: Referral rates decreased from 138 per 1000 person-years in 1987, to 84 per 1000 person-years in 2001. Age differences in referral rates were similar in both surveys. Compared with 1987, more boys than girls were referred to specialists. The overall likelihood of a condition being referred decreased from 8.0 per 100 episodes in 1987 to 6.5 per 100 episodes in 2001. Reasons for referral had also changed by 2001, particularly for the ear, nose, and throat (ENT) specialist and ophthalmologist. Moreover, referral rates for acute otitis media, refractive disorders, and vision problems decreased two- to fourfold in 2001. CONCLUSION: Presently, Dutch general practitioners tend to manage more health problems themselves and refer less young people to specialists. PMID- 15527612 TI - Participation in general practice health screening by people with multiple sclerosis. AB - We examined the effect of mobility on health screening for people with multiple sclerosis. General practice records were searched for blood pressure and cholesterol measurements, lifestyle advice, cervical smears, and mammograms. Blood pressure measurement decreased with decreasing mobility (P <0.001). Lifestyle advice was also related to mobility (P <0.01), with those with a moderate disability most likely to receive lifestyle advice. Overall, wheelchair users received fewer preventative services. Findings were similar for men and women. PMID- 15527613 TI - Effectiveness of computerised rehabilitation for long-term aphasia: a case series study. AB - Seven participants with long-standing aphasia following cerebrovascular accident were serially recruited to a case series study where language therapy was delivered at home and monitored via the Internet. All participants improved in word finding, and four improved in general communication. PMID- 15527614 TI - Professional and patient perspectives of NICE guidelines to abandon maternal monitoring of fetal movements. AB - Over the past 30 years, fetal movement counts have been recommended to women in the second half of pregnancy as a way of monitoring fetal wellbeing and providing an early warning of fetal distress. However, guidance from the National Institute of Clinical Excellence (NICE) recommends abandoning this. Evidence is reviewed to show that the chance of preventing physical damage to the fetus is indeed low. The activity of monitoring movements has been favoured by the majority of women. The new NICE guidance is useful to clarify professional understanding of the limitation of counting fetal movements, but women who notice decreased movements will still need referral for human factors. PMID- 15527616 TI - Response to "A seemingly ineffective study on menopausal memory problems". PMID- 15527615 TI - The effectiveness of topical preparations for the treatment of earwax: a systematic review. AB - BACKGROUND: Earwax is a common problem in both primary and secondary care. There is uncertainty as to the most effective topical treatment. AIM: To assess the evidence concerning the efficacy of topical preparations used for treating earwax. DESIGN OF STUDY: Systematic review and meta-analysis. METHOD: Searching for randomised controlled trials (RCTs) of relevant studies. Classification of preparations into three groups, enabling pooling of data and meta-analysis. RESULTS: Of the 18 RCTs included in the review, four were judged to be of high quality. Fifteen preparations including saline and plain water were studied. Oil based and water-based preparations were equally effective at clearing earwax without syringing (odds ratio [OR] = 0.9, 95% confidence interval [CI] = 0.4 to 2.3) and facilitating successful syringing (OR = 1.0, 95% CI = 0.6 to 1.6). A non water-, non-oil-based preparation appeared more effective than an oil-based preparation at both clearing earwax without syringing, and facilitating successful syringing. Immediate syringing after application of a preparation may be as effective as using eardrops for several days and delaying syringing. CONCLUSIONS: On current evidence, there is little to choose between water-based and oil-based preparations; non-water-, non-oil-based preparations appear promising at both clearing earwax and facilitating successful syringing, but further large trials are needed. Although immediate ear syringing is effective and convenient for patients, it may be less cost-effective than using eardrops and perhaps avoiding syringing. Most of the evidence regarding such a common and time-consuming problem is not of high quality. PMID- 15527617 TI - Patient-centredness. PMID- 15527618 TI - Infrared ear thermometry. PMID- 15527619 TI - Dangerous jobsworths. PMID- 15527620 TI - The sessional phenomenon. PMID- 15527621 TI - Saving money on hernia repairs? PMID- 15527623 TI - Evidence in practice--number 3: Cox 2 inhibitors. PMID- 15527624 TI - Bariatric medicine without surgery is like nephrology without dialysis. PMID- 15527626 TI - Psychiatric diagnoses and psychiatric treatment among bariatric surgery candidates. AB - BACKGROUND: This study investigated the psychiatric diagnoses and psychiatric treatment histories of 90 bariatric surgery candidates. METHODS: Prior to surgery, all participants completed the Weight and Lifestyle Inventory, the Questionnaire on Eating and Weight Patterns, and the Beck Depression Inventory II. Participants also underwent a behavioral/psychological evaluation with a psychologist, which reviewed responses to the measures and provided further assessment of participants' psychiatric status. The evaluation also was used to produce a recommendation on the patients' psychological appropriateness for surgery. RESULTS: Almost two-thirds of patients received a psychiatric diagnosis, the most common of which was major depressive disorder. Nearly two-fifths of all participants, and more than half of those given a psychiatric diagnosis, were engaged in some form of psychiatric treatment at the time of the evaluation. Nevertheless, 64% of patients were unconditionally approved for surgery; 31% were recommended for additional psychiatric or nutritional counseling prior to surgery. Three patients were not recommended for surgery. CONCLUSION: Results of this study provide important information on the preoperative psychiatric status and treatment histories of bariatric surgery candidates. Given the increasing population of bariatric surgery patients, evaluation of patients' preoperative psychiatric status may play an important role in maximizing successful postoperative outcomes. PMID- 15527627 TI - Bariatric surgery worldwide 2003. AB - BACKGROUND: There is a world epidemic of overweight, obesity, and morbid obesity, encompassing 1.7 billion people. Bariatric surgery today is the only effective therapy for morbid obesity. METHODS: E-mail requests for information were sent to the presidents of the national societies of the 31 International Federation for the Surgery of Obesity (IFSO) nations, or national groupings, plus Sweden. Responses were tabulated; calculation of relative prevalence of specific procedures was done by weighted averages. RESULTS: Responders were 26 of 32 (81%) for the general questions and 24 of 32 (75%) for the question on specific operative percentages. In the year 2002-2003, 146,301 bariatric surgery operations were performed by 2,839 bariatric surgeons; 103,000 of these operations were performed in USA/Canada by 850 surgeons. The earliest start date for bariatric surgery was 1953 in the USA; IFSO was founded in 1995. In the year 2002-2003, 37.15% of operations were open; 62.85% laparoscopic. The 6 most popular procedures by weighted averages were: laparoscopic gastric bypass, 25.67%; laparoscopic adjustable gastric banding, 24.14%; open gastric bypass, 23.07%; laparoscopic long-limb gastric bypass, 8.9%; open long-limb gastric bypass, 7.45%; and open vertical banded gastroplasty, 4.25%. Pooling open and laparoscopic procedures, relative percentages were: gastric bypass, 65.11%; gastric banding, 24.41%; vertical banded gastroplasty, 5.43%; and biliopancreatic diversion/duodenal switch, 4.85%. Categorizing into restrictive/malabsorptive, purely restrictive, and primarily malabsorptive, the relative distribution of procedures was 65.11%, 29.84%, and 4.85%, respectively. The number of countries performing gastric banding was 23 (95%), gastric bypass 21 (88%), vertical banded gastroplasty 19 (79%), and biliopancreatic diversion/duodenal switch 16 (67%). Purely restrictive procedures were performed in 24 (100%) of the countries, restrictive/malabsorptive in 21 (88%), and primarily malabsorptive in 18 (75%). CONCLUSIONS: Bariatric surgery is expanding exponentially to meet the global epidemic of morbid obesity. Operative procedures in bariatric surgery are in flux and specific geographic trends and shifts are evident. Yet, of the patients qualifying for surgery, only about 1% are receiving this therapy--the only effective treatment currently available. PMID- 15527628 TI - Preoperative low energy diet diminishes liver size. AB - BACKGROUND: A limited view of the gastro-esophageal area in obese patients is often aggravated by an enlarged liver due to fatty infiltration. Preoperative decrease in liver size would help surgeons, particularly those not used to working with morbidly obese patients. METHODS: 50 morbidly obese patients booked for laparoscopic gastric banding undertook a 2-week, preoperative low energy liquid diet. Ultrasound measurements of the left lobe of the liver and body analysis were undertaken at the start of the diet, and again at the conclusion of the diet (preoperatively). Changes in liver size were compared to body analysis changes. RESULTS: There was a highly significant decrease in liver size in the 2 weeks, which correlated with BMI and weight loss. There was no correlation with fat loss. No large left lobe of the liver was encountered at surgery nor caused any problem in any patients with successful preoperative weight loss. CONCLUSIONS: Preoperative restriction of dietary energy will reduce liver size, and is accurately predicted by associated weight loss. PMID- 15527629 TI - Laryngoscopy and morbid obesity: a comparison of the "sniff" and "ramped" positions. AB - BACKGROUND: The effect of patient position on the view obtained during laryngoscopy was investigated. METHODS: 60 morbidly obese patients undergoing elective bariatric were studied. Patients were randomly assigned into one of two groups. In Group 1, a conventional "sniff" position was obtained by placing a firm 7-cm cushion underneath the patient's head, thus raising the occiput a standard distance from the operating-table while the patient remained supine. In Group 2, a "ramped" position was achieved by arranging blankets underneath the patient's upper body and head until horizontal alignment was achieved between the external auditory meatus and the sternal notch. Following induction of general anesthesia, tracheal intubation was performed using a Video MacIntosh laryngoscope. The laryngoscopy and intubation sequences were recorded onto videotape. Three independent investigators, unaware as to which position the patient had been in at the time of tracheal intubation, then viewed the videotape and assigned a numerical grade to the best laryngeal view obtained. RESULTS: The "ramped" position improved the laryngeal view when compared to a standard "sniff" position, and this difference was statistically significant (P=0.037). CONCLUSION: The "ramped" position is superior to the standard "sniff" position for direct laryngoscopy in morbidly obese patients. PMID- 15527630 TI - Course of metabolic syndrome following the biliopancreatic diversion of Larrad. AB - BACKGROUND: The authors assessed the effect of Larrad's biliopancreatic diversion (BPD) on the main components of the metabolic syndrome. PATIENTS AND METHODS: Plasma concentrations of glucose, insulin, total cholesterol (TC), HDL and LDL cholesterol, triglycerides, LDL/HDL and TC/HDL ratios, and blood pressure and body weight were retrospectively evaluated in 40 patients 3-6, 12, 24 and 60 months after undergoing BPD for morbid obesity with metabolic syndrome. RESULTS: 3-6 months after BPD, glycemia and insulinemia had normalized in 97.5% of the patients and remained stable over the following 5 years. Over this period of 3-6 months to 5 years following BPD, total and LDL cholesterol levels fell by 45.2% and 53.1%, respectively. From 12 months onwards, triglyceride levels decreased appreciably, dropping by 57.4% at 5 years. HDL cholesterol concentrations failed to vary significantly or increased to normal levels in patients showing low initial values. At 5 years, high blood pressure had resolved in 75% of patients and the amount of excess weight lost was 65.5% (+/-14.6). No patient required reversal of the BPD due to severe gastrointestinal or metabolic complications. CONCLUSIONS: Technically adapted to the patient's weight, the Larrad BPD effectively stabilizes the main components of the metabolic syndrome. The BPD has low morbidity rate and should be considered a therapeutic option for patients who do not respond to medical treatment. PMID- 15527631 TI - Impact of age on long-term complications after biliopancreatic diversion. AB - BACKGROUND: The aim of the study is to evaluate the importance of age on the mid- and long-term results and complications after biliopancreatic diversion (BPD). METHODS: Our study comprises 132 morbidly obese patients who underwent Scopinaro BPD from February 1995 to April 2001, with follow-up from 24 to 96 months. The patients, 53 males (40%) and 79 females (60%), with mean preoperative BMI 50.2 (35.4-81.5), and mean age 42 (20-65), were divided in 4 groups. Group A age 20 35, 43 patients; Group B age 36-45, 33 patients; Group C age 46-55, 31 patients and Group D age >55, 25 patients. Incidence of long-term specific complications after BPD were analyzed, including protein malnutrition, reversals, anastomotic ulcer, and incisional hernia. RESULTS: Mean postoperative BMI was similar in all Groups. After 60 months the following BMI values were observed. Group A 30.8, Group B 34.9, Group C 35.9, Group D 32. Incidence of long-term complications were not significantly different (chi(2)) in the 4 Groups, and were respectively: protein malnutrition 6.9%, 12.1%, 6.4%, 16.0%; anastomotic ulcer 11.6%, 9%, 6.4%, 16.0%; reversal 2.3%, 9.0%, 1.32%, 8.0%; ventral hernia 34.8%, 45.4%, 54.8%, 32.0%. CONCLUSIONS: From the preliminary results, it appeared that the incidence of the complications was higher in group D (>55 years old), whereas group C (46 55 years old) showed a lower complication rate. However, the prevalence of complications in all groups was not statistically different on chi(2) analysis. No age limit for bariatric surgery could be determined from the age ranges studied. PMID- 15527632 TI - Duodenal switch is a safe operation for patients who have failed other bariatric operations. AB - BACKGROUND: The incidence of morbid obesity and its surgical treatment have been increasing over the last few years. With this increase, there has been a rise in the number of patients who have had less than desirable outcome after bariatric operations. We perform the duodenal switch (DS) in patients for whom other weight loss surgical procedures have failed, because of inadequate weight loss, weight regain or significant complications, such as solid intolerance or dumping syndrome. METHOD: From November 1999 to March 2004, 46 revisional surgeries were performed at our institution. The data was prospectively collected and reviewed, based on a number of parameters. Operative details, perioperative morbidity, and results are reported. RESULTS: 46 patients had their original bariatric surgical operation revised to DS. This resulted in complete resolution of their presenting complaints. The %EWL was 69% at the time of publication, with a mean lapsed time of 30 months. We had no mortality. Anastomotic leak occurred in 4 patients, 2 in our first 8 patients. We also noted that the majority of the patients were not aware of all the surgical procedures available to them at the time of their original operation. CONCLUSION: In patients in whom gastroplasty, gastric bypass or both have failed to provide adequate weight loss, or worse have resulted in complications, DS can be performed as a safe revisional operation. The revision of other failed bariatric operations to DS results in both weight loss and resolution of the complications. PMID- 15527633 TI - Factors determining conversion from laparoscopic to open Roux-en-Y gastric bypass. AB - BACKGROUND: Conversion from laparoscopic to open Roux-en-Y gastric bypass (RYGBP) is expensive and time-consuming. METHODS: Data from our first 1,000 laparoscopic RYGBP was entered into a database (Minnesota Database-Bariatric, Exemplo Medical). All patients met NIH criteria for bariatric surgery. RESULTS: 41 (4.1%) of 1,000 consecutive lapRYGBPs were converted to open. Patients requiring conversion to open surgery, analyzed for predictors, revealed: 1) BMI, waist size, and weight all were significantly greater in patients converted to open bypass; 2) Gender: 9 of 109 males (8.3%) and 32 of 891 females (3.6%) were converted (Fischer's exact test, P=0.035); 3) Average age of patients converted was 44.9 compared to 41.3 in the lap group (P=0.02); 4) Conversion was required for 12 large livers (1 palpable preop, 7 had diabetes, 7 had NASH or steatosis); 5) 10 conversions for mechanical/technical reasons - 6 for inability to distend abdominal wall and/or manipulate instruments due to thickness of wall, and 2 due to loss of instruments in abdomen; 6) 9 required conversion for adhesions (2 from previous cholecystectomies with biliary leaks, and 1 from previous transverse colectomy; 7) 4 visceral injuries required conversion (2 stomach, 1 small bowel, 1 esophagus); 8) 3 hemorrhages from spleen with blood loss over 1300 ml required conversion (1 spleen removed, 6 minor not requiring open conversion); 9) 3 conversions were for anomaly/disease (1 malrotation of colon, 1 ovarian teratoma, and 1 intra-thoracic stomach). CONCLUSION: Steatohepatitis, diabetes mellitus, adhesions from various causes, previous bile leaks, large waist size, BMI, and weight are predictors for conversion to open surgery. PMID- 15527634 TI - The use of bovine pericardial strips on linear stapler to reduce extraluminal bleeding during laparoscopic gastric bypass: prospective randomized clinical trial. AB - BACKGROUND: A prospective comparison was conducted of extraluminal bleeding following gastric transection with or without staple-line reinforcement by dehydrated bovine pericardium (Peri-Strips Dry--PSD) during laparoscopic Roux-en Y gastric bypass (LRYGBP). METHODS: From January 2001 to September 2003, 98 consecutive morbidly obese patients underwent LRYGBP. Patients were randomly allocated to 2 groups according to the use (Group A, n= 50) or not (Group B, n= 48) of Peri-Strips Dry. In both groups, mortality, intra- and postoperative early and late complications, operating-time, number of hemostatic clips used, blood transfusion and any specific event directly related to the prosthetic material were prospectively evaluated. Data were expressed as mean +/- SD except as otherwise indicated. Statistical analysis was done by means of Student t-test and Fisher exact test. P-value cut off for statistical significance was set at 0.05. RESULTS: Intra- and postoperative mortality were absent. Intra-operative methylene blue test was positive in 6/48 (12.5%) of Group B patients (P<0.001). Overall laparotomic conversion was 3/98 patients (3.1%). One/48 Group B patient was converted because of unsatisfactory exposure and one for linear stapler misfire. One/50 Group A patients was converted for short gastric vessels bleeding during dissection. No patients were re-operated or transfused because of extraluminal bleeding. Mean number of clips used was significantly lower in Group A patients (5 vs 23, P<0.001). The operating-time was significantly less in Group A patients (120+/-60 vs 220+/-100 minutes, P<0.01). CONCLUSIONS: Gastric staple lines reinforced with Peri-Strips Dry result in a significant reduction in the number of Endo-clips used and prevent bleeding. A dry operating field was obtained, and operating-time was significantly reduced. No adverse events could be related to the use PSD. PMID- 15527635 TI - Routine abdominal drains after laparoscopic Roux-en-Y gastric bypass: a retrospective review of 593 patients. AB - BACKGROUND: The authors reviewed the benefits of routine placement of closed drains in the peritoneal cavity following laparoscopic Roux-en-Y gastric bypass (LRYGBP). The purpose of the study was to determine whether routine closed abdominal drainage provides diagnostic and therapeutic advantages in the presence of complications such as bleeding and leaks. MATERIALS AND METHODS: The medical records of 593 consecutive patients who had undergone LRYGBP from July 2001 through May 2003 were retrospectively reviewed. In all cases, antecolic antegastric LRYGBP was performed. Two 19-Fr Blake closed suction drains were left in place, one at the gastrojejunostomy and the other at the jejunojejunostomy. The incidence of bleeding and leaks was reviewed, and the utility of the drains relative to diagnosis and management was evaluated. RESULTS: Bleeding presented in 24 patients (4.4%); in 8, the diagnosis was based on increased sanguinous output from the drain and decreased hematocrit. None of the patients with intraabdominal bleeding required reoperation. Of the 10 patients (1.68%) who presented with leaks, the diagnosis was made within 48 hours postoperatively in 5 patients (50%), based on the characteristics of the drain output. Nonoperative management with drainage and total parenteral nutrition was accomplished in 5 (50%) of the 10 patients with leaks. There was no mortality in the series. CONCLUSION: The routine use of abdominal drains after LRYGBP appears to be beneficial. Drains allowed early diagnosis of complications and in most cases, the successful treatment of leaks. When bleeding is suspected or documented, appropriate volume replacement therapy is mandatory to maintain adequate hemodynamic parameters. Drain output may orient the surgeon to take preventive measures such as discontinuing anticoagulation and early fluid resuscitation. In this series, in most cases the bleeding spontaneously stopped and no further surgical management was required. PMID- 15527636 TI - Fasting plasma ghrelin concentrations 6 months after gastric bypass are not determined by weight loss or changes in insulinemia. AB - BACKGROUND: Ghrelin is a gastric peptide with potent orexigenic effects. Circulating ghrelin concentrations are increased in obese subjects, but increase after weight loss. However, in patients undergoing Roux-en-Y gastric bypass (RYGBP), a decrease in ghrelin levels has been reported. The effect of comparable weight loss induced by either adjustable gastric banding (AGB), RYGBP or conventional dietary treatment (Conv) on ghrelinemia was studied. METHODS: 24 matched obese male patients in whom similar weight loss had been achieved by either AGB (n=8), RYGBP (n=8) or Conv (n=8) were studied before and 6 months after treatment start. The independence of ghrelin concentrations from body mass index (BMI) and weight loss was further analyzed in a group of patients with total gastrectomy (TtGx, n=6). RESULTS: Comparable weight loss after 6 months exerted significantly different effects on plasma ghrelin concentrations, depending on the procedure applied (AGB: 424.6 +/- 32.8 pg/ml; RYGBP: 131.4 +/- 13.5; Conv: 457.3 +/- 18.7; P<0.001). Without significant differences in body weight and BMI, patients who had undergone the RYGBP exhibited a statistically significant decrease in fasting ghrelin concentrations, while the other two procedures (AGB and Conv) showed a weight loss-induced increase in ghrelin levels. Despite significant differences in BMI between RYGBP and TtGx patients after 6 months (31.9 +/- 2.2 vs 22.0 +/- 0.7 kg/m(2), respectively; P<0.05), both groups showed similar ghrelin concentrations. CONCLUSION: The reduction in circulating ghrelin concentrations in RYGBP patients after 6 months of surgery are not determined by an active weight loss or an improved insulin-sensitivity but rather depend on the surgically-induced bypass of the ghrelin-producing cell population of the fundus. PMID- 15527637 TI - Management of incidental ovarian tumors in patients undergoing gastric bypass. AB - BACKGROUND: Ovarian disease is common in obese women and is usually not screened during routine preoperative evaluation in patients undergoing bariatric surgery. Consequently, surgeons may encounter previously undiagnosed adnexal tumors during bariatric operations. The aim of this study is to report our experience with incidental ovarian tumors in patients during Roux-en-Y gastric bypass (RYGBP). METHODS: Prospectively collected data on all consecutive patients undergoing RYGBP for clinically significant obesity from July 1998 to September 2003 were reviewed for patients with incidental gynecological tumors. Details of operative treatment and outcomes are reported herein. RESULTS: 460 women underwent RYGBP during the study period. 52 (11%) had a previous hysterectomy and/or bilateral oophorectomy and were excluded from the study. 12 ovarian masses, median tumor size 11 cm (range 4-65 cm) were found in 10 women (2.5%) during 6 open and 4 laparoscopic RYGBPs. Mean age was 40+/-9 years and mean BMI was 58+/-12 kg/m(2). Resection of 9 benign cystic lesions and 2 malignant lesions was undertaken. One patient with polycystic ovary syndrome did not undergo resection. The RYGBP was completed in all but 2 patients who, by appearance and intraoperative frozen biopsy, had evidence of malignancy. No additional morbidity resulted from the added gynecological procedure. Based on these results, an algorithm for the treatment of incidental gynecological tumors is suggested. CONCLUSIONS: Although infrequent, incidental ovarian tumors may be discovered in patients undergoing bariatric surgery, emphasizing the importance of thorough exploration of the abdominal cavity. Consultation with a gynecologist is warranted in most instances, and treatment should be on a patient-by-patient basis, especially in women of child-bearing age. PMID- 15527638 TI - Elevated serum parathormone after Roux-en-Y gastric bypass. AB - BACKGROUND: Abnormalities in calcium and vitamin D metabolism are observed early after gastric bypass, whereas clinical or biochemical evidence of metabolic bone disease might not be detected until many years after the procedure. The aim of the present study was to evaluate the impact of bariatric surgery on bone metabolism determined on the basis of postoperative laboratory changes in calcium, phosphorus, magnesium, alkaline phosphatase and parathormone (PTH) levels. METHODS: 110 patients submitted to Roux-en-Y gastric bypass (RYGBP) were followed after surgery, and the following parameters were determined: intact PTH molecule (PTHi; chemiluminescence), alkaline phosphatase (colorimetric method), ionic calcium (selective electrode), phosphorus and magnesium (colorimetric method). RESULTS: Elevated serum PTHi levels were observed in 29% of the patients and hypocalcemia in 0.9% from the 3rd postoperative month and afterwards (3 to 80 months after surgery). CONCLUSION: There is a need for careful evaluation of bone metabolism and for routine calcium replacement after RYGBP. PMID- 15527639 TI - Morbid obesity and gastric bypass surgery: biochemical profile. AB - BACKGROUND: Morbid obesity (MO) and the pathologies associated with it constitute an important public health problem, accounting for 7% of the health expenditure in industrialized countries. An important percentage of this expense is attributed to the different biochemical tests performed in these patients, who suffer from several metabolic derangements. We evaluated the basic biochemical abnormalities in MO patients and their reversibility by weight loss after gastric bypass, to standardize the surveillance of the different metabolic abnormalities in obese patients. METHODS: By a retrospective analysis on 125 patients operated in our hospital, we evaluated anthropometric and biochemical data before and 1, 3, 6, 12 and 24 months after gastric bypass. RESULTS: Preoperatively hyperinsulinemia, hyperglycemia, dyslipidemia and hypertensive disease were present, and began to improve 1 and 3 months after surgery (although not significantly) and significantly at 6, 12 and 24 months after it. We also observed deficient protein nutrition and a deficiency of micronutrients both before bypass and during the follow-up. CONCLUSION: After gastric bypass, a marked decrease in insulin occurred, with normalization of blood pressure and the biochemical parameters associated with the metabolic syndrome. We propose a biochemical follow-up protocol for MO patients. PMID- 15527640 TI - Results of silastic ring vertical gastroplasty more than 6 years after surgery: analysis of a cohort of 214 patients. AB - BACKGROUND: Among the different techniques of surgical treatment for morbid obesity, silastic ring vertical gastroplasty (SRVG) is an alternative, effective and easily reproducible technique. The aim of this study is to evaluate a cohort of patients >6 years after SRVG for morbid obesity. METHODS: From 1991 to 1996, 273 consecutive patients were eligible for SRVG. The evaluation criteria included weight loss, evolution of co-morbidities, long-term morbidities, satisfaction of patients and quality of life. RESULTS: Among these 273 patients, 1 patient died in the postoperative period (0.4%). Postoperative morbidities occured in 27 patients (10%). The long-term follow-up involved 213 patients (78%). Late postoperative complications consisted of outlet stoma stenosis (14%), staple-line dehiscence (5.6%) and incisional hernia (8.5%). 23 patients (10%) needed a re-do operation. Co-morbidities drastically improved. BMI fell from 45.3 to 30.7. Failure of SRVG was statistically associated with male gender and super-obese patients. 69% of the patients were satisfied, and 73% would recommend this operation. CONCLUSION: SRVG is very effective in a selected group of morbidly obese patients. PMID- 15527641 TI - Is laparoscopic adjustable gastric banding a day surgery procedure? AB - BACKGROUND: Laparoscopic adjustable gastric banding is the least invasive bariatric operation. However, just isolated attempts to perform this procedure as a Day Case have been published. This study highlights some aspects that might contribute to safe patient discharge within 23 hours after LAGB. METHODS: Prospective evaluation of 20 consecutive patients was carried out. Patients were indicated for laparoscopic Swedish adjustable gastric banding (SAGB, Obtech, Ethicon Endo-Surgery) in a private Bariatric center in the first 6 months of 2003. The effect of extensive pre- and immediate postoperative education and psychological support, and information on postoperative health consequences delivered through a multi-disciplinary bariatric team effort, was evaluated, regarding the influence of these facilitators in shortening the length of hospital stay. RESULTS: Mean preoperative BMI of the 20 patients entering the study was 42.3. Mean operating-time was 91 minutes (58-112 min). Time spent on information and education of each patient was 60 minutes in total during the preoperative period. Average postoperative hospitalization was 21 hours. There were no intraoperative or early postoperative complications. Excess weight loss was 44% at 12 months after surgery. CONCLUSION: SAGB performed on a Day Case basis in selected patients who are subjected to intensive pre- and immediate postoperative dedicated education appears to be a feasible alternative. PMID- 15527642 TI - Impact of preoperative teaching on surgical option of patients qualifying for bariatric surgery. AB - BACKGROUND: During the last 5 years, the performance of bariatric operations has doubled via our outpatient obesity clinic. Currently, 52% of the patients presenting for weight loss are interested in bariatric surgery. Gastric banding and Roux-en-Y gastric bypass are the two laparoscopic procedures proposed. The aim of this study was to evaluate the impact of preoperative teaching on the patients' surgical option. METHODS: All the candidates for bariatric surgery were submitted to preoperative teaching and those between February 2001 and December 2002 are the subject of this study. The teaching consisted of 3 weekly interactive 2-hour sessions. During the first session, the patients were asked about the type of operation that they had in mind: gastric banding, gastric bypass, or not yet decided. The same questions were repeated at the end of the third session, with an additional possible answer: no surgery. RESULTS: 297 consecutive patients with a BMI >35 kg/m(2) with at least one severe co morbidity, were submitted to preoperative teaching. 80% of the patients were women. Median age was 41 years. Before teaching, 68 patients (23%) were uncertain, 100 (34%) favored gastric banding, and 129 (43%) wanted a gastric bypass. After education, only 3 patients (1%) remained uncertain, 45 (15%) changed their surgical option, and 27 (9%) declined surgery. The proportion of patients opting for gastric banding decreased from 34% to 20%, whereas those electing bypass increased from 43% to 70%. CONCLUSIONS: Preoperative training provides an informed and better patient selection for bariatric surgery. It helps the patients understand the various surgical options, and makes their decision easier. PMID- 15527643 TI - Current attitudes to the laparoscopic bariatric operations among European surgeons. AB - BACKGROUND: The current attitudes among European bariatric surgeons toward the laparoscopic bariatric operations were examined. METHODS: 150 questionnaires were sent to recognized bariatric surgeons in Europe, and 60% responded. RESULTS: 47% of respondents perform laparoscopic Roux-en-Y gastric bypass (LRYGBP), 81% laparoscopic adjustable gastric banding (LAGB), and 29% laparoscopic biliopancreatic diversion with or without duodenal switch (L-BPD/BPDDS). For BMI <40, 57% of respondents would only perform LAGB, 7% LRYGBP, 2% vertical banded gastroplasty (VBG), 3% L-BPD/BPDDS, and 2% intra-gastric balloon. For BMI 40-50, 43% of respondents prefer LAGB, 11% LRYGBP, 8% VBG, 5% L-BPD/BPDDS, and 33% contemplate several operations. For BMI 50-60, 30% prefer LAGB, 23% LRYGBP, 5% VBG, 16% L-BPD/BPDDS, and 26% tailor each patient's treatment. For BMI >60, 20% prefer LAGB, 24% LRYGBP, 37% L-BPD/BPDDS, 2% VBG, and 17% consider more than one operation. Although important, BMI and patient eating habits are not significant in choosing an operation for 25% of respondents. Interestingly, 39% of the surgeons offer laparoscopic bariatric surgery to so-called pediatric patients (<18). Of these, 76% favor LAGB, 8% LRYGBP, 8% L-BPD and 4% other procedures. CONCLUSIONS: The overall body of respondents prefers laparoscopic procedures. The responses suggest that at lower BMI there is a higher trend for restrictive operations. However, as BMI increases, combined and malabsorptive operations are preferred. At least one-third of surgeons offer bariatric surgery to patients with age <18 years, and here LAGB is greatly preferred. PMID- 15527644 TI - Surgical management of obese patients with eating disorders: a survey of current practice. AB - BACKGROUND: This study examined the current practice of bariatric surgeons and their colleagues regarding patients with binge eating disorder (BED) and night eating syndrome (NES) who present for Roux-en-Y gastric bypass (RYGBP) for obesity. METHOD: We conducted a 9-item internet survey of American Society for Bariatric Surgery (ASBS) members. For each item, the numbers of respondents endorsing each possible response, including "Other" and "Unknown or not applicable," were tabulated, and percentages of the total sample of respondents were calculated. RESULTS: Most respondents' screening process included mental health (82.0%) and nutritional (78.0%) evaluations. Most inquired about binge eating (88.0%) and other eating disturbances (83.3%), while fewer respondents (52.7%) screened for night eating. Management of patients with eating disorders varied widely. For patients with binge eating, 20.0% of respondents proceeded with surgery, 2.7% recommended against surgery, and 27.3% postponed surgery, with the remainder (50.0%) reporting that their management varied. For night eating and other eating disturbances, responses were similarly diverse. Respondents who postponed surgery reported a wide range of estimates of how often patients with eating disorders follow through with treatment for their eating problem and return for surgery: 16% (always/almost always), 36% (usually), 24% (sometimes), 12% (occasionally), and 12% (never/almost never). CONCLUSION: Although bariatric surgeons commonly screen for eating disorders such as BED, there are limited empirical data and no consensus regarding the optimal management of these patients. PMID- 15527645 TI - Increase in obesity and health-care use, from the Health Survey for England, ages 50 to 69, 1998. AB - BACKGROUND: The relationship between obesity and the use of health-care services was assessed in the population between 50 and 69 in England in 1998, where such analysis had not yet been done. METHODS: All data was obtained from the Health Survey for England (HSE), a cross-sectional survey that collected information about the health of a sample of the British population in 1998. Analysis was limited to the 4,102 men and women between 50-69 years with a valid body mass index (BMI) >18.5 kg/m(2). BMI was analysed in relation to use of 4 types of health-care service, and the odds ratio (OR) for use of each of these services was determined for obese people (BMI > or =30 kg/m(2)), compared with normal weight individuals (BMI 18.5-25). RESULTS: There was an association between obesity status and use of outpatient services in a 12-month period (adjusted 0R 1.25; 95% CI, 1.03-1.51) and with > or =2 medical prescriptions (adjusted OR 1.86; 95% CI, 1.51-2.29). No significant relationship was found between obesity and physician (GP) visits or with use of inpatient and/or day-patient services. CONCLUSIONS: Although obesity did not lead to increases in use in all 4 health care measures assessed as it has in some other countries, it does lead to some increases in health-care utilization in England and appears to impose an additional burden on health-care services. With the rising rates of obesity and its co-morbidities, there may be larger stresses on the health-care system in England if efforts to reduce obesity are not put into effect. PMID- 15527646 TI - The evolution of experimental surgery in the field of morbid obesity. AB - BACKGROUND: We reviewed the experimental surgery, related to bariatric surgery in laboratory animals, to consider new lines of research. METHODS: The literature on experimental surgery for morbid obesity since the mid-20th century was reviewed, focusing on existing techniques (malabsorptive, restrictive, mixed and experimental) and their associated metabolic component. RESULTS: In the field of laparoscopy, there is a clear tendency for large laboratory animals such as pigs to be used. These are useful for developing and perfecting techniques. A second area of animal experimentation concentrates on the relationship between metabolism and surgery in order to find improvements in the co-morbidities associated with morbid obesity. A third area of research focuses on manipulating intake via central and vagal control. CONCLUSION: Further studies are needed to combine traditional and recently developed techniques of experimental surgery with the mechanisms that determine the physiopathology, metabolism and regulation of intake of morbidly obese patients. To accurately determine metabolic behavior and avoid drawing conclusions that are not very significant, these studies should be carried out on obese animals and focus on the co-morbidities associated with morbid obesity. PMID- 15527647 TI - Minimally invasive gastric bypass in a morbidly obese patient with myasthenia gravis. AB - Associated or rare diseases, such as myasthenia gravis, introduce a challenge to the perioperative management of severely obese patients undergoing bariatric surgery. We report the surgical management and unique anesthetic approach to a 55 year-old morbidly obese woman with a complex past medical history that included myasthenia gravis, who underwent laparoscopic gastric bypass. Her myasthenia was controlled on pyridostigmine and her greatest concern was the potential need for postoperative mechanical ventilation. While the laparoscopic surgical approach was ideal to reduce pain and the adverse effects on ventilatory mechanics associated with open upper abdominal surgery, a combined inhalational and intravenous anesthetic without muscle relaxants resulted in satisfactory surgical conditions, and allowed for immediate postoperative extubation followed by an uneventful postoperative course. Continued perioperative anticholinesterase administration may have facilitated this successful outcome. We conclude that a diagnosis of myasthenia gravis does not mandate postoperative mechanical ventilation following laparoscopic gastric bypass. PMID- 15527648 TI - Gastric outlet obstruction caused by total band erosion. AB - Laparoscopic adjustable gastric banding for the treatment of morbid obesity has gained widespread popularity because of its simplicity both for the surgeon and more so for the patient. On the other hand, with this procedure there are complications such as band slippage, erosion, balloon problems and tubing problems, which have required reoperations for remedy. Herein we describe a case of band erosion into the stomach causing gastric outlet obstruction. Of particular interest are the clinical appearance and the operative management of this complication. PMID- 15527649 TI - Small bowel obstruction by the silicone tube of the gastric band. AB - A wide range of operations are used today for morbid obesity. Adjustable gastric banding (AGB) is one of the most widespread. Numerous complications after AGB are known, namely gastric perforation, band slippage, penetration of the band into stomach, port disconnection, port-site infection, etc. The authors present a case of small bowel obstruction caused by the intra-abdominal silicone tube of the gastric band in a woman with AGB performed 9 years before, with a very good result and considerable weight loss. She was operated as an emergency, and part of the terminal ileum was found incarcerated around and between the silicone tube and the anterior abdominal wall. Bowel resection for intestinal necrosis, with terminal ileostomy, was performed, followed 1 month later by an end-to-end ileo ileal anastomosis. The patient recovered without sequelae. PMID- 15527652 TI - Association between the rates of multiple maternities. AB - We analyze the association between the rates of multiple maternities. Correlation analysis is suitable if there are at least monotonic relationships between the variables. A decreasing tendency can be observed in the rates of multiple maternities in Sweden and in its 25 counties for the period 1751-1960 and the effect of external variables can be assumed to be monotonic. After 1960, the rates of multiple maternities show marked increases, mainly caused by ovulation stimulants and the introduction of other artificial reproduction techniques. We transformed the triplet and quadruplet rates according to Hellin's law in order to obtain rates on the same scale as the twinning rate. The time trends for the multiple maternities in Sweden as a whole were quite similar. The regional differences in the twinning rates could not be satisfactorily explained by maternal age. Alternative attempts to eliminate the effect of other time dependent factors were to study partial correlation coefficients when the time was kept fixed and the correlation coefficients based on cross-sectional regional data. Both the ordinary and the partial correlation coefficients showed strong regional variation. Cross-sectional analyses gave correlation coefficients similar to the partial correlation coefficients for Sweden as a whole. The variations in the correlation coefficients between the twinning and the triplet rates seem to be caused by other time-dependent factors and the effects of these factors show strong regional variation. After elimination of such factors, the correlation between the twinning and the triplet rates is moderate. PMID- 15527650 TI - Ghrelin and gastric bypass. PMID- 15527653 TI - Should the statistical analyses of twinning-rate data be improved? AB - Every statistical model is based on explicitly or implicitly formulated assumptions. In this study we address new techniques of calculation of variances and confidence intervals, analyse some statistical methods applied to modelling twinning rates, and investigate whether the improvements give more reliable results. For an observed relative frequency, the commonly used variance formula holds exactly with the assumptions that the repetitions are independent and that the probability of success is constant. The probability of a twin maternity depends not only on genetic predisposition, but also on several demographic factors, particularly ethnicity, maternal age and parity. Therefore, the assumption of constancy is questionable. The effect of grouping on the analysis of regression models for twinning rates is also considered. Our results indicate that grouping influences the efficiency of the estimates but not the estimates themselves. Recently, confidence intervals for proportions of low-incidence events have been a target for revived interest and we present the new alternatives. These confidence intervals are slightly wider and their midpoints do not coincide with the maximum-likelihood estimate of the twinning rate, but their actual coverage is closer to the nominal one than the coverage of the traditional confidence interval. In general, our findings indicate that the traditional methods are mainly satisfactorily robust and give reliable results. However, we propose that new formulae for the confidence intervals should be used. Our results are applied to twin-maternity data from Finland and Denmark. PMID- 15527654 TI - Why zygosity of multiple births is not always obvious: an examination of zygosity testing requests from twins or their parents. AB - This paper examines why parents of twins or adult twins themselves request zygosity testing. Of 405 multiples including 8 sets of triplets, the majority (93%) were monozygotic. Age of testing ranged from 0 days to 73 years. About 50% of requests came from parents or twins who were curious about, or expressed a need to be certain of, their zygosity. Other reasons included health concerns (current or future), other twins in the family, and misinformation about zygosity, frequently because of the erroneous assumption that all dichorionic twins are dizygotic. Parents of monozygotic twins may expect their twins to be 'identical' and believe their twins to be dizygotic because of minor phenotypic differences between them. Dizygotic twins like other siblings may share a phenotypic resemblance. Health professionals should be aware that zygosity of multiples may not always be obvious to parents and that accurate knowledge of zygosity may be justified. PMID- 15527655 TI - Differential transmission of parvovirus B19 in a twin gestation: a case report. AB - Maternal infection with parvovirus B19 during pregnancy can cause aplastic anemia in the fetus. Severe anemia may lead to nonimmune hydrops or fetal demise. In the case reported, the demise of one twin was diagnosed by ultrasonography in an asymptomatic 21-year-old para 1-0-2-1 African American at the gestational age of 25 weeks. The deceased twin (A) was grossly hydropic with anasarca, ascites, pleural and pericardial effusions, and a thickened placenta. Parvovirus B19 DNA was found in the amniotic fluid of Twin A using the polymerase chain-reaction technique. Serial scans of Twin B showed normal growth and no evidence of hydrops. The pregnancy was managed expectantly until 29 weeks when delivery was indicated by maternal disseminated intravascular coagulation. Maternal IgM antiparvovirus B19 antibodies were detected at the time of delivery. Antiparvovirus B19 IgM antibodies were not present in Twin B. These serologic studies suggest a recent acute maternal infection and refute such an infection in Twin B. We present a case of differential transmission of parvovirus B19 in a twin pregnancy with in utero death of the infected twin and subsequent maternal disseminated intravascular coagulation. PMID- 15527656 TI - The effect of birthweight on tooth-size variability in twins. AB - Studies indicate that low birthweight (LBW) children display reduced deciduous tooth size but there is little information about permanent tooth size. It has also been shown that dental fluctuating asymmetry (FA) increases in response to various environmental influences, but the relationship between birthweight and FA remains unclear. The aim of this study was to compare tooth size and asymmetry, according to birthweight, in the deciduous and permanent dentitions of a sample of Australian twins. The study sample comprised 436 twins, classified into 2 groups: normal birthweight (NBW > 2500 g) and low birthweight (LBW < or = 2500 g). For each individual it was generally possible to measure maximum mesiodistal crown diameters of both deciduous and permanent central incisors from serial dental models. Correlations were calculated between tooth-size variables and birthweight; subsequently comparisons of tooth size and FA were made between the LBW and NBW samples using Student's t tests. Small positive correlations (around .1) were noted between birthweight and tooth-size variables. There was no evidence of tooth-size reduction in the LBW male sample, but the LBW females displayed tooth-size reduction of approximately 2-3% for both deciduous and permanent incisors, compared to the NBW females. There was no evidence of increased FA in the LBW individuals of either sex. These findings indicate that developing teeth are generally well-protected from developmental disturbances during prenatal and perinatal periods. Further research is needed to clarify the biological basis of an apparently true but weak association between tooth size and birthweight. PMID- 15527657 TI - Growth patterns in young adult monozygotic twin pairs discordant and concordant for obesity. AB - Weight discordance is very rare in monozygotic (MZ) twin pairs; when found, however, such pairs are advantageous in the search for either environmental or epigenetic causes and consequences of obesity. We analyzed the growth patterns of young adult MZ pairs discordant and concordant for obesity. Screening 5 consecutive birth cohorts (1975-1979) of 22- to 27-year-old Finnish twins (the FinnTwin16 study), we found 14 obesity discordant (Body Mass Index [BMI] difference > or = 4 kg/m2) MZ pairs out of 658. Ten pairs participated in clinical studies. Nine concordant pairs (BMI difference < or = 2 kg/m2) were examined as controls. Lifetime measured heights and weights recorded in hospitals and health centers were traced manually. Height development was similar in all the co-twins of both groups. The weight differences between the co-twins of the discordant pairs began to emerge at 18 years leading to an average discordance of 16.4 kg, 5.6 kg/m2 (p for both = .005) at 25.7 years. The heavier co-twin weighed 221 g (p = .066), 1.0 kg/m2 (p = .01) more already at birth than the leaner, but the differences waned by 6 months of age and reappeared only after adolescence. Both the leaner and the heavier co-twins of the discordant pairs weighed more than expected by the singleton reference values (Cole et al., 1998) after 8 years. The concordant co-twins, on the other hand, grew similarly and after 6 months, their mean growth was not distinguishable from the singleton patterns. Young adulthood represents a critical period of gaining weight irrespective of genetic background in this twin sample. PMID- 15527658 TI - Characteristics of a Japanese adult twin database of high school graduates. AB - This paper profiles a unique cohort of adult Japanese twins. The database contains more than 700 twin pairs, aged 18 to 66 years, who are all graduates of the secondary school attached to the faculty of education of the University of Tokyo. This school was established in 1948, when the study of twins was burgeoning in Japan, and about 10 to 20 pairs of twins have been admitted there every year to participate in studies on twins in education and in related projects. The zygosity of all twins was determined carefully on the basis of various sources. Data from the perinatal period to adulthood were linkable using ID numbers. Follow-up surveys in the field of medical genetics were performed in 1985, 1989 and 1999. For the third survey, which was sent and received exclusively by mail, the distribution and collection process was also assessed in detail. The response rate was around 40%, which statistically was influenced mainly by previous participation and sex. The limitation of this cohort is its selection bias concerning socioeconomic status and its imbalance in favor of monozygotic pairs. PMID- 15527659 TI - Pharmacogenetics of nicotine metabolism in twins: methods and procedures. AB - This article describes a pharmacogenetic investigation of nicotine metabolism in twins. One hundred and thirty-nine twin pairs (110 monozygotic and 29 dizygotic) were recruited and assessed for smoking status, zygosity, and health conditions known or suspected to affect drug metabolism. Participants underwent a 30-minute infusion of stable isotope-labeled nicotine and its major metabolite, cotinine, followed by an 8-hour in-hospital stay. Blood and urine samples were taken at regular intervals for analysis of nicotine, cotinine, and metabolites by gas chromatography-mass spectrometry or liquid chromatography-mass spectrometry and subsequent characterization of pharmacokinetic phenotypes. DNA was genotyped to confirm zygosity and for variation in the primary gene involved in nicotine metabolism, CYP2A6. Univariate and multivariate biometric analyses planned for the future will determine genetic and environmental influences on each pharmacokinetic measure individually and in combination with each other, and in the presence and absence of covariates, including measured genotype. When the analyses are completed, this study will result in a more complete characterization of the impact of genetic and environmental influences on nicotine and cotinine metabolic pathways than has heretofore been reported. The approach taken, with its use of a quantitative model of nicotine metabolism, highly refined metabolic phenotypes, measured genotype, and advanced tools for biometric genetic analysis, provides a model for the use of twins in next generation studies of complex drug-metabolism phenotypes. PMID- 15527660 TI - Behavioral inhibition as a function of relationship in preschool twins and siblings. AB - Monozygotic (MZ) twins spend more time with each other and are more genetically alike than are dizygotic (DZ) twins or nontwin siblings and therefore probably experience less diversity in their playmates than DZ twins, who in turn may experience less diversity than other-age siblings. Thus MZ twins may be more inhibited than DZ twins, who may in turn be more inhibited than nontwin siblings. To test this, 205 children (42 MZ twins, 94 DZ twins, and 69 nontwin siblings) were assessed in a playroom laboratory during free play with an unfamiliar peer. Children's inhibition was rated based on latency to touch a toy for the first time and amount of time spent interacting with the other child. Additionally, the Child Behavior Checklist (CBCL) withdrawn scale was used to assess inhibition according to parent report. Behavioral ratings showed that MZ and DZ twins did not significantly differ on the inhibition or withdrawn ratings. DZ twins were significantly more inhibited than were nontwin siblings, according to laboratory ratings. Greater inhibition of twins during the free peer play situation can be explained by their relative lack of experience in playing with children who are less genetically and phenotypically like themselves. Parental ratings showed exactly the opposite pattern, suggesting a rater bias effect. PMID- 15527661 TI - Gender diagnosticity and androgen receptor gene CAG repeat sequence. AB - The gender diagnosticity (GD) approach of Lippa (1995) was used to evaluate the relationship of within-sex differences in psychological masculinity-femininity to a genetic characteristic, the length of a repeated CAG sequence in the X-linked androgen receptor (AR) gene. Previously assessed adult samples in Australia and Sweden were used for this purpose. A weak relationship (correlations in the range .11 to .14) was obtained in both countries. Additional data from adolescent twins from Australia (12-, 14-, 16-year-olds) did not confirm such a relationship at those ages, especially for males. The fact that this sample consisted of twins permitted two kinds of within-pair comparisons: (1) Did the dizygotic twin who had the longer AR sequence have the higher GD score? (2) Was one twin's GD score more highly correlated with the other twin's AR score in MZ than in DZ pairs? The answer in both cases was negative. Clarification of these relationships will require large samples and measurements at additional ages. PMID- 15527662 TI - Investigating the genetic and environmental structure of Cloninger's personality dimensions in adolescence. AB - In this study we examined the genetic and environmental structure of four dimensions from Cloninger's personality system: novelty-seeking (NS), harm avoidance (HA), reward-dependence (RD), and persistence (PS). Although adult twin studies suggest that these personality dimensions are moderately heritable, this is the first twin study of Cloninger's personality dimensions in adolescence--a period marked by significant physiological and social changes. Study participants included 1851 adolescent twins between the ages of 11 and 18 years; 878 complete twin pairs and 95 singleton-responding twins. Subjects were participants in two community-based samples of twins residing in the state of Colorado. Results indicated that cross-sectional mean levels for NS, HA and RD tended to show modest increases across the adolescent years, while PS showed modest mean decreases. Consistent sex differences in means were found only for RD. Univariate biometrical twin models were used to decompose trait variance into genetic and environmental sources. Results indicated that for NS, HA and RD additive genetic influences and unique environmental effects were sufficient to explain the data. PS, however, could be explained by unique and common environmental effects only, with different patterns of common environmental effects for males and females. We found moderate heritability estimates for NS, HA and RD ranging from .28 to .36- with no evidence for sex-limitation in those influences. PMID- 15527663 TI - Genetic and environmental influences on the relative timing of pubertal change. AB - A multicategory item-response theory model was developed to characterize developmental changes in three items relating to the assessment of puberty in adolescent twin girls and boys. The model allowed for the fixed effects of age on probability of endorsing the responses and for the random effects of individual differences on the timing of pubertal changes relative to chronological age. In girls, the model was applied three-wave data on twin pairs (N = 414 female monozygotic [MZ] and 197 female dizygotic [DZ] pairs) and female twins from boy girl pairs (N = 300 twins) from the Virginia Twin Study of Adolescent Behavioral Development. In boys, the data comprised 318 MZ and 185 DZ pairs and 297 male twins from boy-girl pairs. A total of 3172 and 2790 individual twin assessments were available in girls and boys, respectively, spanning ages 8-17 years. The availability of twin data allows the contributions of genes, the shared environment and individual unique environmental experiences to be resolved in the relative timing of pubertal changes. Parameters of the mixed model including fixed effects of age and random effects of genes and environment were estimated by Markov Chain Monte Carlo simulations using the BUGS algorithm for Gibbs sampling. The estimated standard deviation of random differences in the timing of puberty relative to age was 0.96 years in girls and 1.01 years in boys. The estimated intraclass correlations for the relative timing of pubertal changes were 0.99 +/-0.01 in MZ girls, 0.52 +/-0.02 in DZ girls, 0.88 +/-0.04 in MZ boys and 0.44+/-0.02 in DZ boys, indicating a very large contribution of genetic factors to the relative timing of pubertal change in both sexes. Additive genetic factors account for an estimated 96.3+/-3.3% of the total variance in random effects in girls and 88.0+/-3.6% in boys. Shared environmental influences account for 3.6+/-3.4% in girls and 0% in boys. In girls, nonshared environmental effects explain 0.1+/-0.1% of the total residual variance. The comparable figure in boys is 12.0+/-3.6%. PMID- 15527664 TI - Genotyping accuracy for whole-genome amplification of DNA from buccal epithelial cells. AB - We compared the accuracy of genotyping for DNA extracted from lymphocytes to that of DNA amplified from buccal epithelial cells. Amplification was via a rolling circle/phi29 DNA polymerase commercial kit. Paired buccal and lymphocyte DNA samples were available from 30 individuals. All samples were genotyped for 12 SNPs, 5 microsatellites and 2 VNTRs. The accuracy of genotyping (no-call proportions, reproducibility, and concordance) was similar for DNA from lymphocytes in comparison to amplified DNA from buccal samples. If used with caution, these data suggest that rolling-circle whole-genome amplification can be used to increase the DNA mass available for large-scale genotyping projects based on DNA from buccal cells. PMID- 15527665 TI - The consequences of selective participation on behavioral-genetic findings: evidence from simulated and real data. AB - Nonresponse occurs when individuals either have no chance of being included in a study (noncoverage), refuse to take part (unit nonresponse), or fail to give complete information (item nonresponse). The purpose of this article is to test the possible biasing effects of nonresponse on the results of behavioral-genetic studies. Simulations and a real data 'natural' experiment were used to determine the impact of nonresponse on estimates of additive genetic and environmental effects. The simulations used realistic twin-pair correlations and models of nonresponse derived from prior research. The real data 'natural experiment' used data from a nationally representative birth-cohort twin study (E-Risk Study) and compared model results from families who had responded to a mail survey to those from all study cases. Results showed that the primary influence of nonresponse was to attenuate the effect of the shared environment and to inflate estimates of nonshared environment and additive genetic effects. At high levels of nonresponse a spurious nonadditive genetic effect (suggesting genetic dominance) was also found. Study nonresponse was shown to have the potential to bias the findings of behavioral-genetic research. Design and analysis methods that can be used to alleviate this potentially important biasing effect in behavioral-genetic studies are discussed in light of these findings. PMID- 15527666 TI - Power of the classical twin design revisited. AB - Statistical power of the classical twin design was revisited. The approximate sampling variances of a least-squares estimate of the heritability in a univariate analysis and estimate of the genetic correlation coefficient in a bivariate analysis were derived analytically for the ACE model. Statistical power to detect additive genetic variation under the ACE model was derived analytically for least-squares, goodness-of-fit and maximum likelihood-based test statistics. The noncentrality parameter for the likelihood ratio test statistic is shown to be a simple function of the MZ and DZ intraclass correlation coefficients and the proportion of MZ and DZ twin pairs in the sample. All theoretical results were validated using simulation. The derived expressions can be used to calculate power of the classical twin design in a simple and rapid manner. PMID- 15527667 TI - Linkage analysis: principles and methods for the analysis of human quantitative traits. AB - Currently, mapping genes for complex human traits relies on two complementary approaches, linkage and association analyses. Both suffer from several methodological and theoretical limitations, which can considerably increase the type-1 error rate and reduce the power to map human quantitative trait loci (QTL). This review focuses on linkage methods for QTL mapping. It summarizes the most common linkage statistics used, namely Haseman-Elston-based methods, variance components, and statistics that condition on trait values. Methods developed more recently that accommodate the X-chromosome, parental imprinting and allelic association in linkage analysis are also summarized. The type-I error rate and power of these methods are discussed. Finally, rough guidelines are provided to help guide the choice of linkage statistics. PMID- 15527668 TI - Supplementation of a home-based exercise programme with a class-based programme for people with osteoarthritis of the knees: a randomised controlled trial and health economic analysis. AB - OBJECTIVES: To establish the relative effectiveness and cost of providing a home based exercise programme versus home-based exercise supplemented with an 8-week class-based exercise programme. DESIGN: The trial was a pragmatic, single-blind randomised clinical trial accompanied by a full economic evaluation. SETTING: Patients were randomly allocated to either home-based exercise or home exercise supplemented with class exercise programmes. PARTICIPANTS: A total of 214 patients, meeting the American College of Rheumatology's classification of knee osteoarthritis, were selected from referrals from the primary and secondary care settings. INTERVENTIONS: Both groups were given a home exercise programme aimed at increasing lower limb strength, and endurance, and improving balance. The supplemented group also attended 8 weeks of twice-weekly knee classes run by a physiotherapist. Classes represented typical knee class provision in the UK. MAIN OUTCOME MEASURES: Assessments of locomotor function, using a timed score of three locomotor activities, walking pain and self-reported disability with the Western Ontario and McMaster's Universities osteoarthritis index (WOMAC) were made. General health, lower limb strength, range of movement and compliance with exercise were also measured. Patients were assessed before and after treatment, and also at 6- and 12-month follow-ups. The economic evaluation looked at health service resource use and assessed cost-effectiveness by relating differential costs to differences in quality-adjusted life-years (QALYs) based on patients' responses to the EuroQol-5 Dimensions. Data were obtained at baseline, 1 month, 6 months and 12 months through face-to-face interviews and, where appropriate, examination of hospital medical records. RESULTS: Patients from the supplemented group demonstrated significantly greater improvement in locomotor function and decrease in pain while walking at all follow-ups. The supplemented group also demonstrated smaller but significant improvements in balance, strength, WOMAC score, and the physical function and pain dimensions of the Short Form-36. However, not all of these improvements were maintained over the 12-month follow up period. There was no evidence that compliance with the home exercise programme was different or that total costs or mean QALY gains were significantly different between the groups. However, costs were slightly lower and QALY gains slightly higher in the group with the supplementary class-based programme. The economic evaluation suggests that supplemented programmes are likely to be considered cost effective, although there is uncertainty around this estimate, with approximately 30--35% probability that the intervention would not be cost-effective. CONCLUSIONS: The supplementation of a home-based exercise programme with a class based exercise programme led to superior improvement in the supplemented group. These differential improvements were still evident at review 12 months after treatment had ceased. The additional cost of the supplemented group was offset by reductions in resource use elsewhere in the system. Compliance with the home exercise programme did not differ between the groups. Based on this evidence, the supplementation of a home-based exercise programme with an 8-week class-based exercise programme can be confidently expected to produce small improvements in locomotor function and clinically important reductions in pain. It is recommended that future research investigates methods of increasing compliance with home exercise programmes and evaluates the impact of these interventions in the primary care setting, where most patients with knee osteoarthritis are managed. PMID- 15527669 TI - Clinical and cost-effectiveness of once-daily versus more frequent use of same potency topical corticosteroids for atopic eczema: a systematic review and economic evaluation. AB - OBJECTIVES: To assess the clinical and cost-effectiveness of once-daily use of topical corticosteroids versus more frequent use of same-potency topical corticosteroids in the treatment of people with atopic eczema. DATA SOURCES: Electronic databases. Bibliographies of included studies and related papers. Experts in the field. Manufacturer submissions to the National Institute for Clinical Excellence. REVIEW METHODS: Studies were assessed for inclusion according to predefined criteria by two reviewers. Data extraction and quality assessment were undertaken by one reviewer and checked by a second reviewer. Clinical effectiveness data were synthesised through a narrative review with full tabulation of results. RESULTS: One RCT comparing moderately potent corticosteroids, eight RCTs comparing potent corticosteroids and one RCT comparing very potent corticosteroids were included. No RCTs or CCTs of mild corticosteroids were eligible. Most RCTs were of poor methodological quality, although two were judged to be of good quality. The only study that compared moderately potent corticosteroids found no significant difference between once- and twice-daily application. For potent corticosteroids, some statistically significant differences in numbers of patients responding to treatment were identified favouring twice-daily treatment, but these were inconsistent between physician and patient assessment and outcomes selected for analysis. Two studies found a significant improvement in some symptoms with once-daily mometasone furoate compared with twice-daily application of a different active compound, while a third study found no significant differences. One good-quality study favoured twice-daily application of fluticasone propionate ointment, while other studies found no significant difference or an improvement in one symptom but not others. The only study comparing very potent corticosteroids found a statistically significant difference in comparative clinical response in favour of three-times daily treatment, but no difference in number of patients with at least a good response. There appears to be little difference in the frequency or severity of short-term events, however data are limited. No published economic evaluations were identified. Given findings on clinical effectiveness, where outcomes from the comparators are similar, the relative cost-effectiveness of once-daily versus more frequent application of topical corticosteroids becomes a case of cost-minimisation, where the least-cost alternative should be favoured, all else being equal. Topical corticosteroid products included in this review have a wide variation in price; the cost per 30 g/30 ml varies between GBP0.60 and GBP4.88. Specific decisions on the least-cost alternative, between once-daily and more frequent application of products, will be determined by the relative price of the products being compared. Where patients can be appropriately prescribed once-daily treatment of a similarly priced product, a reduction in the quantity of topical corticosteroid used will be expected. However, issues related to pack size for prescribed products and subsequent waste (unused product) could easily erode any potential saving. The potential cost-savings on prescribed products are very small at a patient level; although given the large numbers of patients with atopic eczema, cost savings in theory could be substantial. The presence of specifically marketed 'once-daily' topical corticosteroids, which are relatively expensive (per unit price), may result in additional costs should there be a general recommendation in favour of once-daily use of topical corticosteroids, compared to more frequent use. CONCLUSIONS: The literature is very limited; that available indicates the clinical effectiveness of once-daily and more frequent application of potent topical corticosteroids is very similar, but it does not offer a basis for favouring either option. The cost-effectiveness of once-daily versus more frequent use will depend on the generalisability of the findings to the specific treatment decision and the relative product prices. The trials included in this review generally refer to moderate to severe atopic eczema, whereas most patients have mild disease, and furthermore most of the included trials report on potent topical corticosteroids (eight of 10 RCTs); therefore the generalisability of the findings is limited. Further research is required on the clinical and cost-effectiveness of once-daily versus more frequent use of same potency corticosteroids, specifically on mild potency products for mild to moderate atopic eczema. Outcomes should include quality of life and compliance. PMID- 15527670 TI - Acupuncture of chronic headache disorders in primary care: randomised controlled trial and economic analysis. AB - OBJECTIVES: To determine the effects of a policy of using acupuncture, compared with a policy of avoiding acupuncture, on headache in primary care patients with chronic headache disorders. The effects of acupuncture on medication use, quality of life, resource use and days off sick in this population and the cost effectiveness of acupuncture were also examined. DESIGN: Randomised, controlled trial. SETTING: General practices in England and Wales. PARTICIPANTS: The study included 401 patients with chronic headache disorder, predominantly migraine. INTERVENTIONS: Patients were randomly allocated to receive up to 12 acupuncture treatments over 3 months or to a control intervention offering usual care. MAIN OUTCOME MEASURES: Outcome measures included headache score; assessment of Short Form 36 (SF-36) health status and use of medication at baseline, 3 months and 12 months; assessment of use of resources every 3 months; and assessment of incremental cost per quality-adjusted life-year (QALY) gained. RESULTS: Headache score at 12 months, the primary end-point, was lower in the acupuncture group than in controls. The adjusted difference between means was 4.6. This result was robust to sensitivity analysis incorporating imputation for missing data. Patients in the acupuncture group experienced the equivalent of 22 fewer days of headache per year. SF-36 data favoured acupuncture, although differences reached significance only for physical role functioning, energy and change in health. Compared with controls, patients randomised to acupuncture used 15% less medication, made 25% fewer visits to GPs and took 15% fewer days off sick. Total costs during the 1-year period of the study were on average higher for the acupuncture group than for controls because of the acupuncture practitioners' costs. The mean health gain from acupuncture during the year of the trial was 0.021 QALYs, leading to a base-case estimate of GBP9180 per QALY gained. This result was robust to sensitivity analysis. Cost per QALY dropped substantially when the analysis incorporated likely QALY differences for the years after the trial. CONCLUSIONS: The study suggests that acupuncture leads to persisting, clinically relevant benefits for primary care patients with chronic headache, particularly migraine. It is relatively cost-effective compared with a number of other interventions provided by the NHS. Further studies could examine the duration of acupuncture effects beyond 1 year and the relative benefit to patients with migraine with compared to tension-type headache. Trials are also warranted examining the effectiveness and cost-effectiveness of acupuncture in patients with headache receiving more aggressive pharmacological management. PMID- 15527671 TI - Helicobacter pylori infection and gastric cancer: host, bug, environment, or all three? AB - Helicobacter pylori is a common bacterial pathogen that colonizes the gastric mucosa of over 50% of the world's population. All infected individuals exhibit chronic gastric inflammation, and approximately 1% of patients develop gastric cancers, including adenocarcinomas and mucosal-associated lymphoid tissue lymphomas. In 1994, the World Health Organization International Agency for Research on Cancer classified H. pylori as a type I, or definite carcinogen. Because the prevalence of gastric cancers among H. pylori-infected patients varies between individuals, countries, and geographic areas, H. pylori disease related outcomes are believed to be determined by an interplay between host factors, bacterial factors, and their interaction with the environment. This review highlights recent advances in our knowledge on H. pylori disease pathogenesis, focusing on the role of the host, bacteria, and environment in the development of gastric carcinoma. PMID- 15527672 TI - The pathobiology of eosinophilic gastroenteritis of childhood: is it really the eosinophil, allergic mediated, or something else? AB - Over the past decade clinicians have witnessed a dramatic rise in the prevalence of eosinophilic gastrointestinal diseases. Diverse symptoms, a broad range of endoscopic findings, and varying histopathologic features pose several questions: Do eosinophils represent an allergic response? What mechanisms drive eosinophils to specific mucosal targets? How do eosinophils affect the gastrointestinal tissues? Recent clinical and basic studies are investigating the pathogenesis of eosinophilic gastrointestinal diseases. This review highlights the literature concerning the mechanisms that govern these diseases, with a specific focus on diseases of gastrointestinal columnar epithelia (eosinophilic gastroenteritis and eosinophilic colitis). The roles of specific chemokines, such as eotaxin, and the data supporting the involvement of eosinophil granule proteins in disease states, are discussed. PMID- 15527673 TI - The spectrum of gastric disease associated with Helicobacter pylori and other infectious gastritides. AB - Helicobacter pylori is the most frequent infection of the stomach worldwide. Some of the people infected with H. pylori develop symptoms of dyspepsia that correlate with pathologic evidence of gastritis and peptic ulcers. In addition, H. pylori has been associated with preneoplastic lesions and with two neoplasias: intestinal-type gastric adenocarcinoma and mucosa-associated lymphoid tissue lymphoma. Rarely, gastric pathology can also be caused by other infectious agents, including fungi, other bacteria, parasites, and viruses. This review describes H. pylori-associated pathology and pathologies related to other infectious agents. PMID- 15527674 TI - Is no NSAID a good NSAID? Approaches to NSAID-associated upper gastrointestinal disease. AB - Upper gastrointestinal disease induced by use of nonsteroidal anti-inflammatory drugs (NSAIDs) remains a major problem that affects a broad segment of the population, given the frequent use of these drugs by prescription and over the counter. The emergence of the cyclooxygenase (COX)-2 selective inhibitors (coxibs) has introduced a new option that may result in less upper gastrointestinal disease. Recent large studies have demonstrated this advantage, with the caveat that concurrent use of low-dose aspirin may mitigate this benefit. Unfortunately, the relatively high cost of the coxibs makes them not cost-effective unless confined to certain higher-risk populations. Finally, even newer versions of NSAIDs, such as nitric oxide (NO)-releasing aspirin and the COX inhibiting NO donors, are potential alternatives to traditional NSAIDs with less upper gastrointestinal toxicity. PMID- 15527675 TI - Zollinger-Ellison syndrome revisited: diagnosis, biologic markers, associated inherited disorders, and acid hypersecretion. AB - Despite general awareness of Zollinger-Ellison syndrome (ZES) by most physicians and more than 3000 articles written about it since 1955, the diagnosis of ZES is still delayed for a mean of 5 years. Recent studies show it is being delayed even more with the widespread use of proton pump inhibitors. A number of tumor markers, in addition to assessing serum gastrin, such as chromogranin A, neuron specific enolase, and subunits of chorionic gonadotropin, have been proposed for use in either the diagnosis of pancreatic endocrine tumors, such as gastrinomas, or for assessment of tumor extent and growth. In this article important recent insights into the diagnosis of ZES as well as the clinical usefulness of assessing tumor markers for diagnosis and determination of disease extent and growth are discussed. Approximately 25% of ZES cases are due to multiple endocrine neoplasia type 1 (MEN1). A number of important studies in this group of patients are also reviewed. Finally, almost every patient with ZES has marked gastric acid hypersecretion, and its current treatment as well as the long-term possible side effects are reviewed briefly. PMID- 15527677 TI - New insights into the pathogenesis of inflammatory bowel disease. AB - Several important advances have been made over the past few years that have expanded our knowledge of the immunology of the gut and its complex interactions with commensal organisms. Critical developments in our understanding of the pathogenesis of inflammatory bowel diseases include the discovery of Toll-like receptors and the identification of not one but two susceptibility genes for Crohn's disease. We have furthered our understanding significantly concerning the role of dendritic cells in the development of gut inflammation. In addition, a novel hypothesis suggesting a protective role for helminthic infections is gaining experimental evidence and direct clinical applicability. In this review we summarize these key developments in the pathophysiology of inflammatory bowel disease and attempt to ascribe clinical relevance where applicable. PMID- 15527676 TI - Advances in the genetics of inflammatory bowel disease. AB - Research efforts in the inflammatory bowel diseases have been uniquely successful in identifying genetic linkage regions likely containing susceptibility genes for Crohn's disease and ulcerative colitis. In two of these regions, definitive gene associations have been established, namely for the NOD2/CARD15 gene on chromosome 16 (IBD1) and the OCTN1/SLC22A4-OCT/SLC22A5 genes on chromosome 5q (IBD5), both conferring increased risk for developing Crohn's disease. Recently, significant gene associations have been reported for additional genes, including DLG5, MDR1, and TLR4 as well. The NOD2/CARD15 gene mutations are associated with ileal disease location and a modestly earlier age of onset compared with NOD2/CARD15 wild-type Crohn's disease patients. Future progress in the genetics of inflammatory bowel disease will likely involve systematic phenotyping, including the incorporation of clinical subtypes and novel biomarkers. The ultimate goal of genetic research in inflammatory bowel disease is to identify the earliest biologic pathways that are altered, resulting in disease pathogenesis. Identification of these key pathways will potentially highlight novel therapeutic targets. PMID- 15527678 TI - Relevance of serologic studies in inflammatory bowel disease. AB - The serologic panel for inflammatory bowel disease (IBD) is rapidly expanding. Antineutrophil cytoplasmic antibodies (ANCA) and anti-Saccharomyces cerevisiae mannan antibodies (ASCA) have remained the most widely studied markers, but immune reactivity against a new group of bacterial antigens such as I2, OmpC (outer membrane porin C), and flagellin, has been described in Crohn's disease. Several clinical avenues have been explored, such as the usefulness of serologic markers as screening tools for IBD and in accelerating a diagnosis in patients with indeterminate colitis. Another area of interest is disease stratification. Emerging data suggest there is a diversity of qualitative and quantitative responses to environmental antigens that differs among groups of IBD patients and may be associated with different clinical behaviors. As a result, it may be possible to tailor therapy on the basis of serologic responses. Prospective studies are needed before translating this concept into clinical practice. Clustering of IBD patients into more homogeneous subgroups based on antibody responses may help to unravel the pathophysiology of subsets of IBD. PMID- 15527679 TI - Medical therapy for ulcerative colitis: the state of the art and beyond. AB - Ulcerative colitis (UC) is an idiopathic inflammatory condition of the large intestine. Recent advances in our understanding of the pathogenesis of UC have led to the development of novel treatments for this often debilitating condition. Aside from aminosalicylates and corticosteroids, drugs that have been used for decades in the treatment of UC, biologic agents, in addition to medications targeting specific effector mechanisms involved in the inflammatory cascade, have been used in patients with UC with varying degrees of success. Clinicians have never had as many therapeutic options for UC as they do today. Herein we review the variety of treatment options, both standard and investigational, that are available for patients with UC. PMID- 15527680 TI - Advances in medical therapy for Crohn's disease. AB - Therapeutic research in Crohn's disease has been intensified in recent years. This has led to many novel approaches and insights into the mechanism of action of "classic" drugs. Antibiotics remain valuable but do not offer benefit when used in addition to corticosteroids. Immunomodulators remain the cornerstone for maintenance therapy, although certain corticosteroid-dependent patients can be switched to maintenance therapy with topical steroids. Azathioprine and 6 mercaptopurine remain efficient beyond 4 years in patients with relapses and elevated C-reactive protein in spite of this therapy. Infliximab has shown efficiency in maintenance of active and fistulizing Crohn's disease. In addition, "automatic reinfusion" was found to be superior to "on-demand" treatment. Infusion reactions and loss of response, most often caused by antibodies against infliximab, can be prevented with immunomodulators and corticosteroid infusions before dosing. Such alternative anti-tumor necrosis factor agents as adalimumab or CDP-870 may be less immunogenic. Other biologic agents, such as the anti integrin monoclonal antibody natalizumab, were shown to be effective in maintaining remission and somewhat less so in induction of remission. Finally, much attention is being paid to alteration of the luminal flora with probiotics and helminth ova. Extracorporeal apheresis and even stem cell transplantation were found to be effective in isolated patients, but these therapies warrant further prospective and controlled investigation. PMID- 15527682 TI - VALUE in hypertension treatment. PMID- 15527681 TI - Management of extraintestinal manifestations and other complications of inflammatory bowel disease. AB - The past 18 months have seen many studies of the prevalence, pathogenesis, and treatment of the extraintestinal manifestations of inflammatory bowel disease (IBD). Inhibitors of tumor necrosis factor alpha have shown effectiveness in randomized trials for the treatment of spondyloarthropathies and ocular manifestations. Open-label studies suggest that these agents may be effective for pyoderma gangrenosum as well. The epidemiology of primary sclerosing cholangitis (PSC), and its relationship to IBD, is becoming clearer. Colorectal neoplasia in PSC remains an important clinical problem. Osteoporosis occurs more commonly in IBD, but the relative importance of corticosteroid use versus underlying chronic bowel inflammation as risk factors remains controversial. Chromoendoscopy may be an important means to improve detection of colorectal neoplasia in IBD. Observational studies suggest that prolonged use of aminosalicylates is associated with decreased risk of neoplasia, but data are conflicting. A randomized trial of ursodeoxycholic acid in PSC showed decreased risk of colorectal neoplasia in patients receiving the drug relative to those on placebo. PMID- 15527683 TI - Myocardial metabolism: a new target for the treatment of heart disease? PMID- 15527684 TI - Insulin signaling in heart muscle: lessons from genetically engineered mouse models. AB - The heart is an insulin-responsive organ, and disorders of insulin action, such as diabetes and obesity, can have profound effects on cardiac performance. Insulin signaling influences numerous functions within the heart, such as metabolic substrate preference, cell size, and the response of the heart to ischemia and hypertrophy. Because the systemic consequences of altered insulin action can have significant but indirect effects on the heart, the generation of mice with altered expression of insulin receptors and key components of the insulin-signal transduction pathways in cardiomyocytes have led to interesting and occasionally surprising new insights into the regulation of cardiac biology by insulin. PMID- 15527685 TI - Altered cardiac calcium handling in diabetes. AB - Diabetes results in a cardiomyopathy characterized by reduced contractility that is primarily the result of changes in calcium handling within the myocyte. Because most of the calcium involved in excitation-contraction (EC) coupling is derived from the sarcoplasmic reticulum (SR), it is no surprise that many studies have found a reduction in sarco-endoplasmic reticulum calcium ATPase (SERCA) activity in the diabetic state. In this review, we outline the changes to SR calcium handling in the diabetic state and, through the use of transgenic mice and adenoviral gene therapy, we examine how SR function can be improved by the expression of various proteins that are directly and indirectly involved in calcium handling. Improving SERCA activity plays an important role in ameliorating the contractile phenotype associated with the diabetic state. PMID- 15527686 TI - Energy substrate metabolism in cardiac hypertrophy. AB - Cardiac hypertrophy is a response to long-term pathologic (eg, hypertension) or physiologic (eg, exercise) hemodynamic overload accompanied by changes in energy substrate utilization. The pattern of substrate utilization (or metabolic phenotype) differs dramatically between pathologic and physiologic cardiac hypertrophy with directionally opposite changes in oxidation of fatty acids and glucose and glycolysis. These findings indicate that the metabolic response to long-term alterations in hemodynamic workload is not stereotypical, but is influenced by the nature of the stimulus leading to cardiac hypertrophy. Although the changes in substrate utilization are adaptive, in the case of pathologic stimuli, the changes in metabolism interfere with functional resiliency of the heart to metabolic stress, as occurs during ischemia-reperfusion. The distinct metabolic phenotypes of hearts hypertrophied in response to pathologic or physiologic stimuli are due not only to alteration in expression of metabolic enzymes and proteins, but also to post-translational modulation of metabolic enzymes and proteins. PMID- 15527687 TI - Arterial stiffness and wave reflection in hypertension: pathophysiologic and therapeutic implications. AB - Numerous studies performed in the past decade have shown that increased pulse pressure, an indicator of arterial stiffening, is associated with adverse clinical outcomes. Research is now focused on better defining the anatomic and physiologic determinants of increased pulse pressure. Early work considered aortic stiffening to be a passive, irreversible consequence of aging that was accelerated by hypertension. Recent studies suggest that aortic stiffening is dynamic and reversible and that abnormalities in aortic function may play a primary role in the pathogenesis of hypertension. This review summarizes current work on the genesis of increased pulse pressure and abnormal pulsatile load in hypertension, and underscores areas for future research. PMID- 15527689 TI - Pharmacogenetic study of statin therapy in cholesterol reduction. PMID- 15527688 TI - Angiotensin II and abdominal aortic aneurysms. AB - Abdominal aortic aneurysms (AAAs) have devastating effects on the morbidity and mortality of a large portion of the elderly population. Current therapeutic options for AAAs are limited to surgical approaches, because there are no proven pharmacologic treatments. Recently, there is evolving evidence that angiotensin II (Ang II) participates in the initiation and propagation of AAAs. Animal studies have consistently demonstrated the ability of Ang II to promote the formation of AAAs, although the mechanisms of this effect have not been defined. Further definition of the role of the renin-angiotensin system in AAA formation and progression will identify potential therapeutic strategies for treatment of this disease. PMID- 15527690 TI - Nicardipine or labetalol as short-term treatment for severe hypertension of pregnancy. PMID- 15527691 TI - Hypertension and liver disease. AB - Arterial hypertension is a common disorder with a frequency of 10% to 15% in subjects in the 40- to 60-year age group. Yet most reports find the prevalence of arterial hypertension in patients with chronic liver disease (cirrhosis) much lower. In this review, we consider the alterations in systemic hemodynamics in cirrhosis. The most characteristic findings in cirrhotic patients are vasodilatation with low systemic vascular resistance, increased cardiac output, high arterial compliance, secondary activation of counterregulatory systems (sympathetic nervous system, renin-angiotensin-aldosterone system, neuropituitary release of vasopressin), and resistance to vasopressors. The vasodilatory state is mediated through nitric oxide, calcitonin gene-related peptide, adrenomedullin, and other vasodilators, and is most pronounced in the splanchnic area. This constitutes an effective (although relative) counterbalance to increased arterial blood pressure. Subjects with established arterial hypertension (essential, secondary) may become normotensive during the development of cirrhosis, and arterial hypertension is rarely manifested in patients with cirrhosis, even in cases with renovascular disease and high circulating renin activity. There is much dispute as to the understanding of homeostatic regulation in cirrhotic patients with manifest arterial hypertension. This is a topic for future research. PMID- 15527692 TI - Hypertension management: special considerations in chronic kidney disease patients. AB - It has been estimated that approximately 11% of the US adult population has chronic kidney disease (CKD), and it has been demonstrated that the prevalence of hypertension rises significantly as renal function declines. Even mild CKD significantly increases mortality risk, and cardiovascular disease remains the main cause of death among these patients. Although CKD patients have generally been excluded from trials testing the effect of lowering blood pressure on cardiovascular outcomes, guidelines suggest lowering blood pressure in hopes of reducing cardiovascular mortality and slowing the progression of renal disease. The preferred antihypertensive agents among these patients are drugs that block the renin-angiotensin system. In most hypertensive CKD patients, however, multiple agents are necessary to reach blood pressure targets. In general, diuretics and calcium channel blockers are added subsequently as adjunctive therapy. Hopefully, with increased recognition of the unique aspects of treating hypertension in this population, end-stage renal disease and cardiovascular morbidity and mortality will be delayed or avoided in the millions of patients with CKD. PMID- 15527693 TI - Hypertension in the elderly. AB - High blood pressure, once believed to represent a normal and progressive component of the aging process, is now recognized as a manifestation of structural and physiologic abnormalities of aortic function. Elevated systolic blood pressure and increased pulse pressure unquestionably increase the risk of both fatal and nonfatal cardiovascular events, including stroke, myocardial infarction, and heart failure. Isolated systolic hypertension, defined as a systolic blood pressure >/= 140 mm Hg with a diastolic blood pressure < 90 mm Hg, affects most individuals aged 60 years and older. Several clinical trials have clearly demonstrated that treatment of hypertension significantly reduces the cardiovascular event rate. However, controversy continues as to the choice of antihypertensive agents and combinations of agents. It is both appropriate and necessary to treat elderly hypertensives aggressively to the same target blood pressures identified for younger patients. It is also appropriate to initiate treatment with lower doses of antihypertensive agents and to bring the pressure down more slowly, monitoring for orthostatic hypotension, impaired cognition, and electrolyte abnormalities. PMID- 15527694 TI - Pheochromocytoma: diagnosis and management update. AB - Pheochromocytoma is a rare but extremely treacherous neuroendocrine tumor causing manifestations by secreting catecholamines into the circulation. It is usually fatal if unrecognized. Manifestations can mimic many diseases and cause erroneous diagnoses. Approximately 15% are malignant, 18% extra-adrenal, and 20% familial. Plasma or urinary metanephrines are approximately 98% sensitive for detecting pheochromocytomas. They can be localized by MRI and CT; (131)I- and (123)I metaiodobenzylguanidine (MIBG) are highly specific and 81% to 90% sensitive, respectively. Suspect pheochromocytoma in patients with sustained or paroxysmal hypertension or in normotensives having manifestations of hypercatecholaminemia. Surgical removal is usually curative; chemotherapy and radiotherapy are palliative for malignant pheochromocytomas. PMID- 15527695 TI - Evolution of diagnostic criteria for primary aldosteronism: why is it more common in "drug-resistant" hypertension today? AB - The recent "epidemic" of primary aldosteronism reported in the literature is most likely related to the widespread acceptance that with easy access to accurate measurements of renin and aldosterone, it is no longer necessary to wait until hypokalemia has become profound before embarking on diagnostic testing to attempt to ferret out this most common cause of "essential" hypertension. This is especially true for those who are now classified as "drug resistant" using today's popular drugs, which are particularly ineffective in lowering blood pressure in primary aldosteronism and its variants. Understanding the physiologic consequences of a slowly increasing aldosterone production by autonomous cells will help both the family practitioner and the specialist understand the role of the aldosterone renin ratio (ARR) in the care of the hypertensive patient. In addition, the increasing number of specific genetic mutations that drive sodium retention and lead to low levels of renin activity and familial hypertension must be incorporated into the routine evaluation and care of hypertensive patients and their families. PMID- 15527696 TI - Glucocorticoid excess and hypertension. AB - Cushing's syndrome is a rare disorder characterized by chronic, excess glucocorticoid exposure. Hypertension is one of the most discriminating features of the disease, as it is present in 80% of patients. Patients with Cushing's syndrome have a mortality rate four times that of the general population, most likely secondary to an increased number of cardiovascular risk factors, including hypertension. In this article, we review several pathogenetic mechanisms of glucocorticoid-induced hypertension, including the role of sodium/water and mineralocorticoid excess, as well as involvement of the vasculature and kidney. Although treatment of hypertension with available antihypertensive medications is only moderately successful, after cure of Cushing's syndrome, approximately 30% of patients have persistent hypertension. PMID- 15527697 TI - Evidence-based medicine in pediatric rheumatology: progress or peril. PMID- 15527699 TI - Clinical trials in Kawasaki disease. PMID- 15527698 TI - Defects in the disposal of dying cells lead to autoimmunity. AB - The fast and efficient uptake of dying cells is of main importance to prevent contact of the immune system with intracellular autoantigens. Insufficient clearance of the latter is discussed to drive the humoral autoimmune response in systemic lupus erythematosus. Many adaptor molecules and receptors are involved in the recognition of dying cells. In this paper we focus on the involvement of phosphatidylserine, glycoproteins, and complement and DNaseI in the clearance of apoptotic and necrotic cells, respectively. Furthermore, extracellular danger signals released from necrotic cells are discussed and the uptake process of primary necrotic cells is investigated in detail. Last but not least, the character and origin of clearance defects observed in some systemic lupus erythematosus patients is presented. PMID- 15527700 TI - Systemic manifestations in localized scleroderma. AB - In this paper, the various systemic manifestations reported in localized scleroderma, their incidence, their relationship with systemic sclerosis, and their relationship with other autoimmune or connective tissue diseases will be analyzed. PMID- 15527701 TI - Dyslipoproteinemia and premature atherosclerosis in pediatric systemic lupus erythematosus. AB - While modern treatments for systemic lupus erythematosus (SLE) have resulted in greatly improved long term outcome in children and adults, complications of atherosclerosis have become a major cause of morbidity and mortality. Although children and adolescents with SLE rarely experience adverse cardiovascular events before adulthood, dyslipoproteinemia and early evidence of premature atherosclerosis is present much earlier. Accelerated atherogenesis in SLE is multifactorial, most likely reflecting vascular, immune, and inflammatory changes along with medication effects. The long term complications of cardiovascular disease in childhood lupus present a particularly important target for intervention because of the potential return on investment by significantly lengthening life and improving quality of life over many decades. An ongoing multi-center, randomized, controlled trial, Atherosclerosis Prevention in Pediatric Lupus Erythematosus (APPLE), testing the efficacy of statins in preventing premature atherosclerosis in children and adolescents with SLE will guide future therapeutic intervention. PMID- 15527702 TI - The role of immune tolerance in preventing and treating arthritis. AB - It has become increasingly clear that the innate and adaptive arms of the immune response cooperate in generating autoimmune damage in the pathogenesis of rheumatoid arthritis and juvenile idiopathic arthritis. Treatment targets the immunologic pathophysiology of the disease and is based on regaining immune tolerance. Recently introduced biological agents neutralize or simply block cytokines and their proinflammatory pathways, with favorable clinical outcome. However, major downsides are their lack of specificity and the need of continuous administration to be effective. Possibly, more can be gained from a specific approach. Indeed, recent findings suggest that targeting antigen-specific T cells can reinstate regulatory mechanisms and thus induce immune tolerization. This improved understanding has paved the way to novel immunotherapeutic approaches, some of which will be discussed here. PMID- 15527703 TI - Primary versus secondary antiphospholipid syndrome: is this lupus or not? AB - Antiphospholipid syndrome is a hypercoaguable state characterized by recurrent venous and/or arterial thrombosis and/or pregnancy complications of fetal loss, pre-eclampsia, or eclampsia in the presence of antiphospholipid antibodies. It was first described in the setting of systemic lupus erythematosus and subsequently recognized to also exist as an independent condition and in conjunction with a variety of other autoimmune, infectious, and malignant illnesses. These diseases have been called primary antiphospholipid syndrome and secondary antiphospholipid syndrome. However, the two conditions can have significant overlapping features. This paper reviews the similarities and the differences between the two conditions. PMID- 15527704 TI - What is antiphospholipid syndrome? AB - The description of antiphospholipid syndrome (APS) is vascular thrombosis and/or pregnancy morbidity occurring in persons with antiphospholipid antibodies (aPL). However, because of an unclear etiopathogenesis and the heterogeneous nature of aPL and of aPL-related clinical manifestations, a single, unambiguous definition of APS does not exist. In this paper, we describe a structured approach to APS, discuss the controversies, and offer descriptions of APS that include: an autoimmune systemic disease with a spectrum of (mostly thrombotic) clinical manifestations, an overdiagnosed disease when aPL is present but has no proven direct causative role in the clinical manifestations, and a disease of controversies that requires clarification of etiology and mechanisms and risk stratified controlled clinical studies. PMID- 15527705 TI - Genetics of antiphospholipid syndrome. AB - Antiphospholipid syndrome (APS) is an autoimmune disease characterized by recurrent arterial or venous thrombosis or fetal loss and the presence of antiphospholipid antibodies (aPL). Genetic factors are thought to play a role in the susceptibility to APS. Similar to many other polygenic autoimmune diseases, human leukocyte antigen associations have been reported. The genetics of b(2) glycoprotein I, one of the most representative target antigens of aPL, has been extensively studied. Additional genetic risk factors for the development of thrombosis in patients with aPL have also been discussed. However, the genes involved in APS have not been identified because antigen specificity of aPL and the pathophysiology of APS are highly heterogeneous and multifactorial. Genome wide linkage analysis and larger cohort studies would lead to better understanding of the genes that might be involved in APS. PMID- 15527707 TI - Diagnosis of the antiphospholipid syndrome: how far to go? AB - The past decade has seen an evolution in the way that thrombophilic conditions are diagnosed and understood. This has largely evolved through the detection of single nucleotide polymorphisms in critical regulating proteins that are thought to confer significant structural-functional changes at key points in the coagulation cascade. The antiphospholipid syndrome (APS) is a complex hypercoagulable disorder that as yet defies the possibility of simple, predictive testing. PMID- 15527706 TI - Treatment of the antiphospholipid antibody syndrome. AB - Numerous questions exist regarding the proper management of patients with the antiphospholipid antibody syndrome (APS). Several recent, randomized-controlled trials have been conducted that attempt to answer the fundamental questions of whom to treat, how to treat, and for how long treatment should last. These studies suggest that APS should be categorized by disease manifestations, and these presentations may have different treatment algorithms. A better understanding of the immune mechanisms that govern thrombosis in this syndrome has led to a wide array of innovative treatment approaches that will require meticulous study. PMID- 15527708 TI - Nonalcoholic Steatohepatitis. AB - Nonalcoholic steatohepatitis (NASH) is an important medical condition and there is great public health concern related to its increasing incidence and potential implications for the development of end-stage liver disease. NASH represents a progression beyond simple lipid deposition in the liver parenchyma, requiring histologic evidence for hepatocyte injury such as ballooning degeneration, Mallory bodies, and/or pericellular fibrosis that can potentially lead to progressive liver injury and eventually cirrhosis. It is believed that several insults contribute to the evolution of hepatic injury such as insulin dysregulation, lipid deposition, oxidative free radicals, and lipid perioxidation. Initial treatment protocols for NASH focus on various aspects of injury in an attempt to control insulin imbalances, improve lipid regulation, reduce free radicals, and ameliorate the inflammatory process. No therapy is conclusively beneficial in all individuals, but preliminary data suggest several approaches that hold promise. PMID- 15527709 TI - Treatment of Hepatocellular Carcinoma. AB - Hepatocellular carcinoma is the fifth leading cause of cancer worldwide and its incidence is increasing. Surveillance programs allow doctors to identify patients at early stages of the disease, when the tumor may be curable by radical treatments such as resection, liver transplantation, or local ablation. In the West, these treatments can be applied to 30% to 40% of patients. Resection yields favorable results in patients with single tumors and a well-preserved liver function (5-year survival rate is 60%). Recurrence complicates two thirds of the cases, and there is no effective adjuvant treatment. Liver transplantation is the best treatment for patients with single tumors that are less than 5 cm in diameter and liver failure, or in those presenting with three nodules less than 3 cm, but organ shortage greatly limits its applicability. Long-term survival is expected to be around 50% to 70% at 5 years depending upon the drop-out rate of patients on the waiting list. Chemoembolization and local ablation are the neo adjuvant treatments applied to patients on the waiting list to prevent tumor progression; no controlled study proving their efficacy has yet been published. In nonsurgical candidates, percutaneous treatments (ethanol injection or radiofrequency ablation) are the best therapeutic approach and improve survival in Child-Pugh A class patients with small tumors that achieve initial complete response (5-year survival rate is 40% to 50%). At more advanced stages, chemoembolization, a technique combining intra-arterial chemotherapy and selected ischemia, has shown to slightly improve survival in a meta-analysis of randomized trials. No survival advantages have been demonstrated with intra-arterial or systemic chemotherapy, hormonal compounds, or radiation. New agents, such as inhibitors of the tyrosine kinase receptors of growth factors and antiangiogenic agents, are currently being tested in phase II/III trials. PMID- 15527710 TI - Hepatitis A. AB - Hepatitis A infection is typically transmitted by the fecal-oral route. Symptomatic infection is highly dependent on the age of the patient and usually follows a self-limited course. Once diagnosed, clinical and biochemical follow-up in the outpatient setting is generally appropriate. Treatment aims are to achieve symptomatic relief and to maintain adequate hydration and caloric intake. In patients with more severe disease, hospitalization may be needed to accomplish aggressive symptomatic therapy and close monitoring of liver function tests and mental status. Prompt evaluation for liver transplantation is appropriate in the rare case where fulminant liver failure develops. Given the absence of specific therapy for hepatitis A virus infection, the most important health care intervention is prevention of infection and/or transmission, which can be accomplished with the safe and effective use of immune globulin and commercially available vaccines. PMID- 15527711 TI - Alcoholic Hepatitis. AB - Alcoholic hepatitis (AH) is a common disease associated with significant morbidity and mortality. Most often the diagnosis is suggested by a history of heavy alcohol excess in a patient with features of hepatic decompensation. In its purest form, AH is a histologic diagnosis of acute hepatic inflammation in response to alcohol. The primary objective of treatment for AH is to support long term alcohol abstinence and to achieve adequate nutrition with lifestyle modification; goal setting and education are integral to long-term medical management. Severity at presentation (calculated by way of the Maddrey score) determines outcome. Patients with AH represent a heterogeneous group with regard to severity and pathogenesis, with various therapeutic interventions assessed in patients with severe AH. To date, corticosteroids have been studied most, and despite remaining controversial, warrant a place in the treatment of selected patients. Recent advances in unraveling the aspects of disease pathogenesis in AH have raised the possibility of targeted therapies, such as anti-tumor necrosis factor-a monoclonals and pentoxifylline. Orthotopic liver transplantation is not recommended for patients with severe acute AH, as most have an unclear long-term prognosis in the context of ongoing excess alcohol ingestion at presentation. PMID- 15527712 TI - Acute Liver Failure. AB - Acute liver failure (ALF) is an uncommon medical emergency whose rapid progression and high mortality demand early diagnosis and expert management, including immediate transfer of any potential case to facilities for intensive care and orthotopic liver transplantation (OLT). All patients with ALF must be screened aggressively for acetaminophen toxicity (history, serum levels, "hyperacute" presentation with renal failure), for other drugs, and viral hepatitis; rare causes of ALF should also be considered. After an acetaminophen overdose, N-acetylcysteine must be given as early as possible, preferably in the emergency room, but any patient with ALF should promptly receive N-acetylcysteine if there is suspicion of acetaminophen toxicity irrespective of the time of ingestion. Supportive care for all patients with ALF includes adequate enteral nutrition, aggressive screening and treatment of infection, prophylactic broad spectrum antibiotics, and antifungal agents. Sedation with propofol is given for severe agitation or mechanical ventilation. With advanced coma grades, intensive care is needed with hemodynamic monitoring, ventilatory support, continuous renal replacement for renal failure, and intracranial pressure monitoring. Intracranial hypertension is treated with mannitol and/or acute short-term hyperventilation, but if the patient is refractory to treatment, mild-moderate hypothermia is achieved by a cooling blanket that is continued throughout OLT. Barbiturate coma is only used in refractory cases as the last treatment modality. Seizures are aggressively treated with phenytoin, with additional diazepam as needed. Candidacy and activation for OLT should be completed as early as possible in the course of ALF, especially in "hyperacute" cases such as acetaminophen toxicity. The final decision to proceed with OLT is made when a donor organ becomes available. King's College Hospital criteria for OLT are still the best prognostic assessment for fatal outcome in ALF, but the criteria fail to identify some patients who will die. PMID- 15527713 TI - Nonresponders to Previous Chronic Hepatitis C Treatment. AB - The main strategy governing treatment of chronic hepatitis C is the prevention of future liver complications. There is good evidence that curing hepatitis C infection prevents progression of liver disease and allows histologic regression to occur. Therefore, the primary goal of medical treatment is to cure the viral infection. Combination therapy with peginterferon alfa and ribavirin is the current standard of care; there are no other medical therapies currently available. Those who failed to respond to an earlier version of antiviral therapy should strongly consider treatment with peginterferon/ribavirin if possible. Nearly half of patients who start peginterferon/ribavirin are unable to achieve a sustained disappearance of infection. If there were problems related to dosing or adherence the first time around, it is reasonable to consider re-treating with more aggressive support. Nonresponders to the current therapy who have early stage liver disease can afford to wait until new antiviral agents come along in the next 5 to 10 years. However, physicians should encourage nonresponding patients with advanced fibrosis to consider experimental alternatives in the meantime, provided there is a logical rationale for the treatment proposed. Some re-treatment strategies still aim to cure the hepatitis C virus infection whereas others focus on limiting liver damage. The best candidates for the first strategy are patients who had temporary clearance of the virus during previous treatment and those with hepatitis C virus genotype 2 or 3 infection. Logical candidates for the second strategy are those who already have advanced fibrosis. It is preferable to pursue further attempts at treatment within the framework of a controlled trial. Studies with strong rationales include those investigating high dose peginterferon/ribavirin, long-term peginterferon suppression, potential immune modulators, and potential inhibitors of liver fibrosis. The rationales are weaker for re-treatment with a second brand of peginterferon/ribavirin, daily standard interferon plus ribavirin, and ribavirin monotherapy. PMID- 15527714 TI - Chronic Hepatitis B. AB - Interferon alpha, lamivudine, and adefovir are the three drugs currently approved for the treatment of chronic hepatitis B virus (HBV). There are pros and cons associated with the use of each drug. Individualization of therapy, based upon factors such as patient comorbidities, response to prior therapies, and stage of disease, is recommended. Patients with abnormal liver enzymes, indices of active viral replication (HBV DNA positive hepatitis B early antigen positive) or evidence of necroinflammatory activity on liver biopsy, and compensated liver disease are potential candidates for treatment with interferon, lamivudine, or adefovir. Patients with abnormal liver enzymes, indices of active viral replication (HBV DNA positive HB(e)Ag positive), and decompensated liver disease are candidates for treatment with lamivudine or adefovir. Consideration of liver transplantation should occur concurrently. Interferon alpha treatment results in hepatitis B surface antigen loss and sustained suppression of HBV DNA replication in 30% to 40% of treated patients. Loss of HBsAg occurs in nearly 10% of patients and a higher than expected frequency of HBsAg loss occurs long-term. The main limitation of therapy is the side effects and the need for parenteral administration. Additionally, interferon therapy is not applicable to all patient groups. Lamivudine achieves HB(e)Ag seroconversion in 15% to 20% of patients treated for 12 months, but (HBsAg) loss is rare. Reduction in HBV DNA to undetectable levels (by hybridization assay) during treatment is nearly universal, and histologic improvement is seen in about 55% of patients. The main limitation of lamivudine therapy is the development of drug resistance, which occurs in 20% of patients after 12 months and increases with duration of therapy (55% at 3 years). Adefovir achieves HB(e)Ag seroconversion in 12% of patients treated for 12 months, but HBsAg loss is rare. An average 3.5 log reduction in HBV DNA levels is and histologic improvement occurs in 50% to 60% of patients. It is effective against both wild-type and lamivudine-resistant HBV. The risk of drug resistance is low and estimated to be approximately 2% to 3% after 2 years of treatment. Several new antiviral agents are currently under evaluation in clinical trials. In addition, there are two drugs (tenofovir and emtricitabine) that have been approved for HIV infection and that have anti-HBV activity. In the future, combination therapy for chronic HBV infection can be anticipated. Utilization of two or more anti-HBV drugs would be predicted to enhance efficacy and reduce the likelihood of emergence of drug resistance. PMID- 15527715 TI - Current Treatment for Chronic Hepatitis C. AB - The number one choice for treatment of chronic hepatitis C is the combination of once weekly subcutaneous pegylated interferon plus daily oral ribavirin. The duration of treatment and dose of ribavirin must be tailored to the hepatitis C virus (HCV) genotype. Patients infected with HCV genotype 1 should be treated for 48 weeks with a standard dosage of ribavirin (1000 or 1200 mg/d). This ribavirin dosage regimen is 'off-label' when used in conjunction with pegylated interferon alfa-2b (12 kD). The approved dosage for use in combination with this agent is 800 mg daily. The appropriate duration of treatment and dosage of ribavirin for patients infected with HCV genotype 2 or 3 differs depending on the pegylated interferon that is chosen. It is important to note that the treatment paradigm for these individuals is quickly evolving. When using peginterferon alfa-2a (40 kD) in patients with HCV genotype 2 or 3, the duration of treatment should be 24 weeks in combination with a low dose of ribavirin (800 mg/d). When using pegylated interferon alfa-2b (12 kD) in patients infected with HCV genotype 2 or in patients infected with genotype 3 and hepatitis C virus RNA less than 600,000 IU/mL, the duration of treatment should be 24 weeks. However, recent data suggest such treatment may not be optimal for patients infected with HCV genotype 3 and hepatitis C virus RNA at levels greater than or equal to 600,000 IU/mL; treatment duration may need to be greater than 24 weeks. When using pegylated interferon alfa-2b (12 kD) in patients infected with HCV genotype 3 and high viral load, the optimal dosage of ribavirin appears to be 800 to 1400 mg/d based on bodyweight. PMID- 15527716 TI - Treatment of the Pruritus of Cholestasis. AB - The etiology of the pruritus of cholestasis is unknown. It is inferred that the pruritogen(s) is produced in the liver, excreted in bile, and as a result of cholestasis it accumulates in plasma. It may follow, logically, that the removal of the substance(s) that mediate pruritus leads to its resolution. The problem with this approach, however, is that the substance(s) is unknown; thus, it is not possible to reduce its serum levels specifically. Oral cholestyramine, a resin that is not absorbed, is associated with increased fecal excretion of certain substances, including cholesterol and bile acids. Many patients respond to treatment with cholestyramine with a relief of pruritus, which unfortunately may be temporary, but is well tolerated in general and it seems reasonable to prescribe it as an initial therapy. When pruritus is not relieved by resins, the use of opiate antagonists (eg, naloxone and naltrexone) is supported by data from controlled clinical trials. Butorphanol is an agonist at the kappa opioid receptor and an antagonist at the mu opioid receptor with minimal or absent abuse potential. The use of butorphanol spray in selective patients may be a therapeutic alternative. In uncontrolled observations dronabinol, an agonist at the cannabinoid B1 receptor, and sertraline, a serotonin reuptake inhibitor, have been reported to be associated with the relief of pruritus. The cannabinoidergic and serotoninergic systems participate in the mediation of nociception; therefore, there appears to be a rationale for the use of these drugs to treat pruritus. Data from controlled clinical trials on the use of dronabinol and sertraline, however, are not available at present. PMID- 15527717 TI - How I became a drug "smuggler". PMID- 15527718 TI - Steroid is effective for vestibular neuritis, valacyclovir is not. AB - Methylprednisolone, starting at 100 mg/d and tapering to 10 mg over 3 weeks, is an effective treatment for vestibular neuritis. Valacyclovir (Valtrex) is not effective. PMID- 15527719 TI - Do automated external defibrillators save lives? AB - Automated external defibrillators accompanied by an intensive training program of local volunteers results in a greater number of patients surviving to hospital discharge. The number of additional survivors (14 in 497 community facilities over a 22-month period) is relatively small, however, and the cost of the defibrillators is high, so whether this is the best allocation of healthcare resources remains uncertain. PMID- 15527720 TI - Second x-ray reading not always necessary. AB - Primary care clinicians are able to identify radiographs for which a second reading by a radiologist is unlikely to result in a change in clinical care. The investigators report that no substantial changes in care or episodes of averted patient harm occurred. PMID- 15527721 TI - Fever, weightbearing, sedimentation rate best for diagnosing hip infection. AB - To distinguish septic arthritis from transient synovitis of the hip in children, the following factors are very helpful: a history of fever, inability to bear weight, erythrocyte sedimentation rate higher than or equal to 40 mm/hour, and a peripheral white blood cell count greater than 12,000/mm(3). PMID- 15527722 TI - Omeprazole 20 mg equal to 40 mg for primary care acid-related dyspepsia. AB - Omeprazole (Prilosec) 20 mg is highly effective for the treatment of acid-related dyspepsia. There was no advantage to higher doses, and relapse following the initial 2-week treatment period was common. PMID- 15527723 TI - Influenza vaccine: new recommendations for infants and children aged 6 to 23 months. PMID- 15527724 TI - Consider colonoscopy for young patients with hematochezia. AB - BACKGROUND: Hematochezia is a common complaint in adult patients aged <50 years. Most studies of lower endoscopy for rectal bleeding have concentrated on older patients or have failed to mention the location of lesions. OBJECTIVE: To determine the findings of complete colonoscopy in adults younger than 50 years with rectal bleeding. METHODS: Data were retrieved from medical records and included demographics, indications, endoscopic findings, and histology. Lesions were labeled according to location: proximal to the splenic flexure or distal to (and including) the splenic flexure. Excluded were those with a history of colitis, colorectal cancer, polyps, anemia, significant weight loss, severe bleeding, or strong family history of colorectal cancer. RESULTS: The study included 223 patients with rectal bleeding aged <50 years who had undergone a colonoscopy. Normal findings were recorded for 48 (21.5%). Four (1.8%) were diagnosed with cancer in the distal colon, and 22 (9.9%) were found to have colon adenomas, 6 of whom had proximal adenomas only. Hemorrhoids were present in 135 patients (60.5%). Other findings included colitis, angiodysplasia, diverticulosis, anal fissures, and rectal ulcers. CONCLUSIONS: Colon neoplasms may be present even in younger adults with nonurgent rectal bleeding. Though most findings were benign and located in the distal colon, colonoscopy should be strongly considered for this patient group. PMID- 15527725 TI - Pearly penile lesions. PMID- 15527727 TI - Metformin in type 2 diabetes. PMID- 15527726 TI - Abnormal vaginal discharge: what does and does not work in treating underlying causes. AB - Antifungal medications for intravaginal use have been available in the United States for more than a decade. Women may be inclined to self-diagnose yeast infections with any vaginal discharge or other vulvovaginal symptoms that they deem abnormal. As we saw in the first part of this article, "Abnormal vaginal discharge: Using office diagnostic testing more effectively" (J Fam Pract 2004; 53[10]:805-814), abnormal discharge is more likely to be bacterial vaginosis or no pathogen at all. Potential delay in diagnosis and treatment of a sexually transmitted disease is also a concern. Increasing resistance of Candida sp. to imidazoles is associated with indiscriminate use of over-the-counter products. PMID- 15527728 TI - How should we treat and diagnose obstructive sleep apnea? PMID- 15527729 TI - Nondaily hormonal contraception: establishing a fit between product characteristics and patient preferences. PMID- 15527730 TI - Clinical inquiries. How effective is gastric bypass for weight loss? PMID- 15527731 TI - Clinical inquiries. For knee pain, how predictive is physical examination for meniscal injury? PMID- 15527732 TI - Clinical inquiries. Do steroid injections help with osteoarthritis of the knee? PMID- 15527733 TI - Clinical inquiries. Does acyclovir help herpes simplex virus cold sores if treatment is delayed? PMID- 15527734 TI - Clinical inquiries. How should thyroid replacement be initiated? PMID- 15527735 TI - Clinical inquiries. Is nedocromil effective in preventing asthmatic attacks in patients with asthma? PMID- 15527736 TI - Estimating mortality reduction by comparing survival curves. PMID- 15527737 TI - Internalization and down-regulation of mu opioid receptors by endomorphins and morphine in SH-SY5Y human neuroblastoma cells. AB - The human neuroblastoma cell line, SH-SY5Y, was used to examine the effects of morphine and the endogenous opioid peptides, endomorphin-1 (EM-1) and endomorphin 2 (EM-2), on mu opioid receptor (MOR) internalization and down-regulation. Treatment for 24 h with EM-1, EM-2 or morphine at 100 nM, 1 microM and 10 microM resulted in a dose-dependent down-regulation of mu receptors. Exposure of cells to 10 microM EM-1 for 2.5, 5 and 24 h resulted in a time-dependent down regulation of mu receptors. Down-regulation of mu receptors by morphine and EM-1 was blocked by treatment with hypertonic sucrose, consistent with an endocytosis dependent mechanism. Sensitive cell-surface binding studies with a radiolabeled mu antagonist revealed that morphine was able to induce internalization of mu receptors naturally expressed in SH-SY5Y cells. EM-1 produced a more rapid internalization of mu receptors than morphine, but hypertonic sucrose blocked the internalization induced by each of these agonists. This study demonstrates that, like morphine, the endomorphins down-regulate mu opioid receptors in a dose- and time-dependent manner. This study also demonstrates that morphine, as well as EM 1, can induce rapid, endocytosis-dependent internalization of mu opioid receptors in SH-SY5Y cells. These results may help elucidate the ability of mu agonists to regulate the number and responsiveness of their receptors. PMID- 15527738 TI - The caudal end of the rat spinal cord: transformation to and ultrastructure of the filum terminale. AB - Contrary to the current belief, the spinal cord of the rat does not terminate with the conus terminalis (CT), but its basic components (central canal, gray matter, white matter) continue in the filum terminale (FT). Proceeding caudally in the conus terminalis, first the motoneuron cell column discontinues in the ventral horn. More caudally the dorsal horns separate from the intermediate zone, and discontinue. The ensuing filum terminale consists of the slit-like central canal lined by ciliated ependymal cells, the periventricular gray matter and the peripheral white matter. Uniform small size neurons and glial cells populate the gray matter. Ultrastructural analysis revealed various types of axodendritic and axosomatic synapses as well as fine unmyelinated axons. The white matter consists mainly of myelinated nerve fibers. The neuronal components of the filum terminale, if they occur also in the human spinal cord, should be involved in the diagnosis and treatment of various diseases, e.g. tethered spinal cord syndrome, vascular malformations and disraphysm. PMID- 15527739 TI - Vestibulosympathetic reflex mediates the pressor response to hypergravity in conscious rats: contribution of the diencephalon. AB - To investigate the mechanism of arterial pressure (AP) regulation during hypergravity, the AP response to gravitational force was examined in conscious rats and the AP was found to increase, depending on the degree of gravity load induced by centrifugation. At 20 s after application of 2, 3, or 5 G, the AP increased by 9+/-2, 20+/-3, or 24+/-3 mm Hg, respectively. The AP increase during first 60 s was suppressed by vestibular lesion or pretreatment with hexamethonium, suggesting that the vestibular system and sympathetic nerve system be involved, respectively, in the afferent and efferent pathways. To further examine the central pathway of this response, Fos expression in the brain was examined after exposure to 5 G for 90 min. Intense Fos expression was seen in the medial vestibular nucleus, paraventricular hypothalamic nucleus, autonomic nuclei in the brain stem in intact rats, but not in rats with vestibular lesion. To examine the involvement of the diencephalic nuclei in this pressor response, AP was measured under hypergravity in rats with midcollicular transection. In these rats, the AP change was minimal at 2, 3, and 5 G, indicating that nuclei rostral to the transection level were involved in the pressor response. These results indicate that output from the vestibular system project to the diencephalon, and activation of diencephalic nuclei is indispensable to the pressor response via the sympathetic nerve system. PMID- 15527740 TI - Effect of vitamin E treatment on N-methyl-D-aspartate receptor at different ages in the rat brain. AB - A comparative study using membrane homogenate binding, autoradiography, and Western blot assays was carried out to determine the age-related changes in N methyl-D-aspartate (NMDA) receptors in 4-, 12- and 24-month-old male Wistar rats, treated or not with vitamin E. Vitamin E treatment was 20 mg/kg i.p. daily for 15 days. [(3)H] 5-methyl-10,11-dihydro-5H-dibenzo (a,d) cycloheptan-5,10-imine maleate (MK-801) binding was significantly increased in all areas studied (cortex and hippocampus) at all ages when rats received this treatment. A Western blot study in vitamin-E-treated rats and their controls did not reveal significant differences in the amounts of NR2A, an NMDA receptor subunit widely distributed in the brain mainly in cortex and hippocampus. We conclude that the effect of vitamin E on NMDA receptors is largely age independent. Previous reports and our data have described the presence of age-dependent NMDA receptor changes. The effect of vitamin E in aging is considered to be mediated by free radical scavenging, but from our data, we conclude that this mechanism is not relevant for age-dependent NMDA receptor changes. Our results also support that age or vitamin E treatment have no relevant effects on NR2A subunit, at least until 24 months in rats. PMID- 15527741 TI - 5-HT3A receptor subunits in the rat medial nucleus of the solitary tract: subcellular distribution and relation to the serotonin transporter. AB - The 5-hydroxytryptamine 3 (5HT3) receptor is a serotonin-gated ion channel implicated in reflex regulation of autonomic functions within the nucleus of the solitary tract (NTS). To determine the relevant sites for 5-HT3 receptor mediated transmission in this region, we used electron microscopic immunocytochemistry to examine the subcellular distribution of the 5HT3 receptor subunit A (5HT3A) in relation to the serotonin transporter (SERT) in the intermediate medial NTS (mNTS) of rat brain. The 5HT3A immunolabeling was detected in many axonal as well as somatodendritic and glial profiles. The axonal profiles included small axons and axon terminals in which the 5HT3A immunoreactivity was localized to membranes of synaptic vesicles and extrasynaptic plasma membranes. In dendrites and glia, the 5HT3A immunoreactivity was located on the plasma membranes or in association with membranous cytoplasmic organelles. The dendritic plasmalemmal 5HT3A labeling was prominent within and near excitatory-type synapses from terminals including those that resemble vagal afferents. The 5HT3A-labeled glial processes apposed 5HT3A-immunoreactive axonal and dendritic profiles, some of which also contained SERT. Terminals containing 5-HT3A and/or SERT were among those providing synaptic input to 5HT3A-labeled dendrites. Thus, 5HT3A has a subcellular distribution consistent with the involvement of 5-HT3 receptors in modulation of both presynaptic release and postsynaptic responses of mNTS neurons, some of which are serotonergic. The results further suggest that the neuronal as well as glial 5HT3 receptors can be activated by release of serotonin from presynaptic terminals or by diffusion facilitated by SERT distribution at a distant from the synapse. PMID- 15527742 TI - Effects of intrathecal BAM22 on noxious stimulus-evoked c-fos expression in the rat spinal dorsal horn. AB - The effects of bovine adrenal medulla 22 (BAM22), a cleaved product of proenkephalin A, were investigated on the noxious stimulus-evoked expressions of spinal c-fos-like immunoreactivity (FLI). Heat (51 degrees C) applied to the tail evoked FLI predominantly in laminae I-II of the sacral spinal cord. Intrathecal (i.t.) BAM22 at a dose of 7 nmol decreased the expressions of the heat-evoked FLI by 68%, 64% and 56% in laminae I-II, III-IV and V-VI, respectively, and the decrease pattern was comparable to that induced by i.t. morphine (10 mug). Naloxone (1 mg/kg, i.p.) significantly enhanced the heat-evoked FLI in laminae III-VI, prevented the morphine-induced inhibition, and decreased the potencies of BAM22 in laminae I-II and V-VI by 23-40%. Higher dose of naloxone (10 mg/kg, i.p.) also partially reduced the BAM22-induced suppression. Following intraplantar injection of formalin (2.5%), FLI neurons were preferentially distributed not only in laminae I-II but also in laminae III-IV and V-VI of segments L4-L5. Pretreatment with BAM22 (7 nmol, i.t.) reduced the formalin evoked FLI neurons by 72%, 61% and 58%, in laminae I-II, III-IV and V-VI, respectively. Naloxone (1 mg/kg. i.p.) enhanced the formalin-evoked expressions of FLI in laminae III-VI and decreased the potencies of BAM22 by 22-38% in laminae I-II and V-VI. The present study provided evidence at a cellular level showing that opioid and non-opioid effects of BAM22 on nociceptive processing in acute and persistent pain models were associated with modulation of noxious stimulus-evoked activity of the spinal dorsal horn neurons. PMID- 15527743 TI - [3H]DPDPE binding to delta opioid receptors in the rat mesocorticolimbic and nigrostriatal pathways is transiently increased by acute ethanol administration. AB - Dopaminergic transmission in the mesolimbic and nigrostriatal pathways plays a key role in the reinforcement mechanisms and brain sensitivity to ethanol, respectively. Ethanol reinforcement and high alcohol drinking behaviour have been postulated to be partially mediated by a neurobiological mechanism involving the ethanol-induced activation of the endogenous opioid system. Activation of opioid neural pathways by ethanol may include alterations in the processing, release and/or the receptor binding of opioid peptides. The aim of this work was to investigate the effects of acute ethanol administration on delta opioid receptors in the rat mesocortical, meso-accumbens and nigrostriatal pathways by quantitative receptor autoradiography, using [(3)H] (2-D-penicillamine, 5-D penicillamine)-enkephalin as radioligand. A significant increase in [(3)H] (2-D penicillamine, 5-D-penicillamine)-enkephalin binding was observed in the substantia nigra pars reticulata 1 h after ethanol treatment. Two hours after drug exposure, ligand binding was significantly increased in the frontal and prefrontal cortices, the core and shell regions of the nucleus accumbens, and in the anterior-medial and medial-posterior regions of the caudate-putamen. In contrast, ligand binding was significantly decreased in the posterior region of the caudate-putamen 30 min after ethanol administration. The observed effects may reflect ethanol-induced changes in ligand binding affinity and/or in receptor density. Our results suggest that transitory changes in delta opioid receptors with different kinetic patterns may be involved in ethanol reinforcement and brain sensitivity to the drug. Ethanol-induced delta receptor up- and down regulation mechanisms may participate in modulation of dopaminergic transmission in the mesocorticolimbic and nigrostriatal pathways. PMID- 15527744 TI - Serotonin 2A receptors modulate tail-skin temperature in two rodent models of estrogen deficiency-related thermoregulatory dysfunction. AB - Menopause-associated thermoregulatory dysfunction, including hot flushes and night sweats, is effectively treated by hormonal therapies that include estrogens. Evidence suggests that estrogen regulates serotonin 2A (5-HT(2A)) receptor expression and that 5-HT(2A) receptors are involved in thermoregulation. Therefore, the role of 5-HT(2A) receptors in thermoregulation was assessed in two rat models of ovariectomy-induced thermoregulatory dysfunction. The first model is based on measurement of the tail-skin temperature (TST) increase following naloxone-induced withdrawal in morphine-dependent ovariectomized (OVX) rats (MD model), while the second model relies on telemetric assessment of diurnal TST changes in ovariectomized rats (telemetry model). Treatment with a 5-HT(2A/2C) receptor agonist, (-)-2,5-dimethoxy-4-iodoamphetamine hydrochloride (DOI), prevented the naloxone-induced TST increase in the MD model and restored normal active-phase TST in the telemetry model. The selective 5-HT(2A) receptor antagonist, MDL-100907, had no effect on the naloxone-induced flush when administered alone in the MD model, but it decreased DOI's ability to abate the flush. In the telemetry model, MDL-100907 attenuated the DOI-induced decrease in active-phase TST. Interestingly, MDL-100907 increased TST in both models when given alone, with the TST increase occurring prior to the naloxone-induced flush in the MD model. To evaluate the role of central nervous system (CNS) 5-HT(2A) receptors in TST regulation, DOI was administered in combination with a known peripheral 5-HT(2A/2C) receptor antagonist, xylamidine, in the MD model. Xylamidine had no effect on DOI's ability to abate the naloxone-induced flush. These results indicate that activation of central 5-HT(2A) receptors restores temperature regulation in two rodent models of ovariectomy-induced thermoregulatory dysfunction. PMID- 15527745 TI - Differential c-Fos expression in the newborn lamb nucleus tractus solitarius and area postrema following ingestion of colostrum or saline. AB - Visceral stimuli and the gut-brain axis play a crucial role in the control of ingestion even in the neonate. The aim of this study was to assess the neuronal activation in the nucleus tractus solitarius (NTS) and the area postrema (AP) following nutritional and non-nutritional stimulations. Lambs received a single gastric infusion of colostrum or saline at 5% birth weight or were sham infused. Infusion of either liquid led to c-Fos-like immunoreactivity (c-FLI) in the NTS and AP. Differences were observed along the sections of the NTS rostro-caudal axis according to the nature of the stimulation, suggesting a specificity of certain afferents and/or NTS areas for nutritional or non-nutritional signals. In the AP, the neuronal activation induced by colostrum was much higher than that induced by saline. A higher number of TH-immunoreactive cells were activated following colostrum infusion, suggesting a specific involvement of the catecholaminergic pathway in the treatment of meal-related stimuli. In spite of functional convergence, the two medullary structures observed responded differently according to the stimulation, indicating a complementary role in the integration of visceral signals. PMID- 15527746 TI - Changes in S100B cerebrospinal fluid levels of rats subjected to predator stress. AB - Predator stress is a type of psychogenic stress induced by an innate recognition of threat. S100B, a calcium-binding protein secreted by astrocytes, has been associated with neurotrophic or neurotoxic action in several neuropsychiatric disorders. It has been recently demonstrated that serum S100B levels in rats are increased after stress by immobilization [S. Scaccianoce, P. Del Bianco, G. Pannitteri, F. Passarelli, Relationship between stress and circulating levels of S100B protein, Brain Res. 1004 (2004) 208-11]. This study aimed to measure cerebrospinal fluid (CSF) S100B in rats after an acute stress situation, which is induced by exposure to a predator. S100B was measured in CSF and in hippocampal and cortical slices by ELISA. Forty-three male Wistar rats, aged 70 days, were randomly assigned to handled (control) or stressed groups (exposed to a cat for 5 min). CSF and brain tissue were removed 1 or 24 h after the procedures. Rats exposed to the cat demonstrated a biphasic change in CSF S100B levels. An increase was observed at 1 h after cat exposure, and a decrease was observed 24 h later, although this was not accompanied by changes in S100B content in hippocampus or cerebral cortex. The effectiveness of the stressor used was confirmed by increased freezing response (during cat exposure) and increased anxiety in the plus maze test (1 h after cat exposure). These results indicate that CSF S100B is changed by stress, reinforcing the possibility that this protein is involved in the adaptive response to stress and/or in secondary neuropsychiatric disorders. PMID- 15527747 TI - Dexamethasone induces different wheel running activity than corticosterone through vasopressin release from the suprachiasmatic nucleus. AB - During the analysis of wheel running activity, we found that corticosterone (1 mg/100 g BW) injection decreased wheel activity, while dexamethasone (0.1 mg/100 g) increased the activity. To clarify the functional differences between corticosterone and dexamethasone, we measured Arg-vasopressin (AVP) release from the suprachiasmatic nucleus (SCN) slice culture in vitro and AVP coding mRNA in the SCN in vivo. The corticosterone (0.2 and 2 microg/ml, final concentration in medium) decreased the AVP release, while it increased by dexamethasone (0.2 and 2 microg/ml). An AVP mRNA in the SCN was decreased by both corticosterone (1 mg/100 g) and dexamethasone (0.1 mg/100 g). The differences in wheel activity by corticosterone and dexamethasone are discussed from the changes of AVP in the SCN. PMID- 15527748 TI - Substance P promotes sleep in the ventrolateral preoptic area of rats. AB - Substance P (SP) has been characterized as an excitatory neurotransmitter and/or neuromodulator in the peripheral and central nervous systems. It is involved in mediating various biological functions such as smooth muscle contraction, neuronal excitation, and pain transmission. Although Lieb et al. reported that intravenous infusion of SP into healthy men led to an increase of paradoxical sleep latency and time awake, little is known about the function and target of SP on sleep-wakefulness cycle in the central nervous system. The ventrolateral preoptic area (vLPO) plays an important role in modulation of sleep-wakefulness cycle. The present study investigated the effect of SP on sleep-wakefulness cycle in the vLPO of rats. Slow wave sleep (SWS) was enhanced after SP was microinjected into bilateral vLPO, while SP receptor antagonist, N-acetyl-l tryptophan 3,5-bis(trifluoromethyl)-benzyl ester, led to the opposite effect. The effect induced by SP was blocked by U73122, a phospholipase C inhibitor. In addition, 3-mercaptopropionic acid, a glutamic acid decarboxylase inhibitor that inhibits gamma-aminobutyric acid (GABA) synthesis and release, blocked the SP induced sleep-promoting effect in the vLPO. These results indicate that SP has sleep-promoting effect in the vLPO possibly by GABAergic neurons. PMID- 15527749 TI - Effect of subdiaphragmatic vagotomy on bacterial DNA-induced IL-1beta expression in the mouse hypothalamus. AB - We investigated whether bacterial DNA (CpG-DNA)-induced IL-1beta expression in the mouse hypothalamus is mediated via afferent vagus nerve. Subdiaphragmatic vagotomy did not modify the CpG-DNA (i.p.)-induced IL-1beta expression in the hypothalamus, indicating that CpG-DNA-induced IL-1beta expression is independent of the afferent vagus nerve originating from the subdiaphragmatic organs. On the other hand, we observed the Toll-like receptor 9 mRNA expression in the hypothalamus, suggesting that circulating CpG-DNA acts directly in the brain. PMID- 15527750 TI - Temporal patterns of the embryonic intermediate filaments nestin and vimentin expression in the cerebral cortex of adult rats after cryoinjury. AB - The expression of two embryonic intermediate filaments, nestin and vimentin, in the rat brain at days 0 (control), 1, 4, 7 and 14 post-cryoinjury was studied to elucidate their roles in brain injury. Western blot analysis showed that both nestin and vimentin expressions in the ipsilateral cerebral cortex were significantly increased at 4 and 7 days post-cryoinjury, and were decreased at day 14 after cryoinjury. Immunohistochemistry showed that there were few nestin- and vimentin-positive cells in the cerebral cortex in normal controls. On days 4 and 7 post-injury, abundant glial cells in the periphery of the lesion were immunostained for nestin and/or vimentin; only vimentin was detected in the majority of inflammatory cells in the core lesion. These findings suggest that nestin and vimentin contribute to the repair of brain injury through the migration of activated cells and the formation of a glial scar. PMID- 15527751 TI - The ventral uvula of the mouse cerebellum: a neural target of ethanol and vestibular stimuli. AB - The present study demonstrates that mice exposed to vertical translation stimulation exhibit a distinct parasagittal pattern of Fos-immunoreactive (Fos IR) granule cells in the ventral uvula of the cerebellum. This pattern is identical to one produced by acute ethanol treatment. In contrast, yaw stimulation produces an entirely different pattern in this same region of the cerebellum. Similar results are obtained in the light or in total darkness. These results suggest an anatomical and functional organization within the granule cells of the ventral uvula that may be a common neural substrate for some effects of ethanol and particular vestibular stimuli. PMID- 15527752 TI - Regulating cell survival by controlling cellular energy production: novel functions for ancient signaling pathways? AB - Cell survival is maintained by growth factors and critically depends on sufficient energy supply. New evidence suggests that a rise in cellular energy production is not merely a homeostatic response to increased demand but subject to regulation by extrinsic factors. The mechanisms operating in this control are largely enigmatic. Work on transformed cells identified direct targeting of glycolytic enzymes by signaling proteins as one possibility. But mitochondrial oxidative phosphorylation and biogenesis may also be subject to regulation by growth and survival factors. Both, positive and negative regulators of cell survival impinge on the processes of cellular energy production to regulate growth and survival versus death. PMID- 15527753 TI - Rendezvous in a membrane: close packing, hydrogen bonding, and the formation of transmembrane helix oligomers. AB - The interaction of transmembrane alpha-helices is promoted by a detailed fit between two helical surfaces, which results in close packing and van der Waals interactions of amino acid side chains between two helices. Recent studies additionally indicate an important role of hydrogen bonding for mediating and stabilizing transmembrane helix-helix interactions. The interplay between close packing and electrostatic interactions in influencing the specificity of helix helix interactions on the one hand and the stability of an existing interaction on the other hand is still unknown. Here, we suggest that close packing mainly determines the specificity of a helix-helix interaction, whereas hydrogen bonding is important for stabilization of a preformed helix dimer. PMID- 15527754 TI - Evidence for copper homeostasis function of metallothionein (MT3) in the hyperaccumulator Thlaspi caerulescens. AB - Metallothioneins chelate metals and consequently may be a control point of metal homeostasis. Homologous to type 3 metallothioneins, TcMT3 cDNA was identified in the Cd/Zn hyperaccumulator, Thlaspi caerulescens. TcMT3 amino acid sequence showed modifications in the Cys positions when compared with its Arabidopsis orthologue. A structural model established that the MT3 carboxyterminal domain is similar to the beta domain of animal metallothioneins and predicts a smaller cavity to chelate metals for A. thaliana than for T. caerulescens. Functional testing in yeast and Northern blot analysis added further evidence for adaptative variations of MT3 for the maintenance of Cu homeostasis in a metal hyperaccumulator. PMID- 15527755 TI - Protein-protein interaction between monomers of coliphage HK022 excisionase. AB - Excisionase (Xis) is an accessory protein that is required for the site-specific excision reaction of the coliphages HK022 and lambda. Xis binds in a strong cooperative manner to two tandem binding sites (X1 and X2) located on the P arm of the attachment (att) sites on the phage genome. As a result of crosslinking experiments in vivo and in vitro of Xis-overexpressing cells, by gel filtration of purified Xis and by FRET analyses we show that Xis monomers of HK022 interact and form dimers that are not dependent on the single Cys residue of the protein and on the presence of DNA. The formation of the dimers may explain the strong binding cooperativity of Xis to its sites on DNA. PMID- 15527756 TI - An LEA group 3 family member is involved in survival of C. elegans during exposure to stress. AB - In order to establish a functional role for late embryogenesis abundant (LEA) proteins in response to stress conditions in Caenorhabditis elegans, we silenced the expression of an LEA (Ce-lea-1) gene and determined the survival of worms under stress conditions. Ce-lea-1 transcription was induced during dehydration of C. elegans dauer juveniles. Following partial silencing of Ce-lea-1 transcription, we demonstrated a specific and significant reduction in worm survival during induction of desiccation, osmotic and heat stress. Together, these results establish a functional role for Ce-lea-1 in stress survival of C. elegans and suggest that Ce-lea-1 may function as a component that is common to the responses to the examined stress conditions. PMID- 15527757 TI - UDP-galactose 4-epimerase from Kluyveromyces fragilis: existence of subunit independent functional site. AB - UDP-galactose 4-epimerase from Kluyveromyces fragilis is a stable homodimer of 75 kDa/subunit with non-covalently bound NAD acting as cofactor. Partial proteolysis with trypsin in the presence of 5'-UMP, a strong competitive inhibitor, led to a degraded product which was purified. Results from SDS-PAGE, size-exclusion (SE) HPLC and ultracentrifugation indicated its monomeric status and size between 43 and 45 kDa. 'Two-step assay' with UDP-glucose dehydrogenase as coupling enzyme in the presence of NAD ensured epimerase activity of the monomer. The possibility of transient dimerization of monomeric epimerase during catalysis was excluded by SE HPLC in the presence of excess substrate and NAD. This truncated enzyme retained catalytic site related properties like Km for UDP-galactose, 'NADH-like coenzyme fluorescence' and 'reductive inhibition' similar to its dimeric counterpart. Reversible reactivation of the monomer was achieved up to 95% within 3 min from 8 M urea induced unfolded state, indicating that the catalytic site could form independent of its quaternary structure. Equilibrium unfolding between 0 and 8 M urea indicated that the monomer was less stable compared to the dimer. Chemical modification of amino acids and reconstitution with etheno-NAD suggested that the architecture around the catalytic site of the monomer was conserved. Specific modification reagents further confirmed that the cysteine residues required for catalysis and coenzyme fluorophore reside exclusively on a single subunit negating a 'subunit sharing model' of its catalytic site. PMID- 15527758 TI - Heme binds to and inhibits the DNA-binding activity of the global regulator FurA from Anabaena sp. PCC 7120. AB - Heme is an iron-containing cofactor that aside from serving as the active group of essential proteins is a key element in the control of many molecular and cellular processes. In prokaryotes, the family of Fur (ferric uptake regulator) proteins governs processes essential for the survival of microorganims such as the iron homeostasis. We show that purified recombinant FurA from Anabaena sp. PCC 7120 interacts strongly with heme in the micromolar range and this interaction affects the in vitro ability of FurA to bind DNA, inhibiting that process in a concentration-dependent fashion. Our results provide the first evidence of the possible involvement of heme in the regulatory function of cyanobacterial Fur. PMID- 15527759 TI - Cloning and functional expression of invertebrate connexins from Halocynthia pyriformis. AB - Unlike many other ion channels, unrelated gene families encode gap junctions in different animal phyla. Connexin and pannexin genes are found in deuterostomes, while protostomal species use innexin genes. Connexins are often described as vertebrate genes, despite the existence of invertebrate deuterostomes. We have cloned connexin sequences from an invertebrate chordate, Halocynthia pyriformis. Invertebrate connexins shared 25-40% sequence identity with human connexins, had extracellular domains containing six invariant cysteine residues, coding regions that were interrupted by introns, and formed functional channels in vitro. These data show that gap junction channels based on connexins are present in animals that predate vertebrate evolution. PMID- 15527760 TI - The novel gene fad104, containing a fibronectin type III domain, has a significant role in adipogenesis. AB - A novel gene named fad104 (factor for adipocyte differentiation-104), whose expression level quickly increased in the early stage of adipogenesis, was isolated and characterized. The deduced amino acid sequence of fad104 revealed the possible presence of a fibronectin type III domain and transmembrane domain. The expression of fad104 was detected in adipocyte differentiable 3T3-L1 cells but not observed in the non-adipogenic cell line NIH-3T3. Moreover, the ability of 3T3-L1 cells to differentiate declined with the knockdown of fad104 by RNA interference, strongly indicating that fad104 functions as a positive regulator of adipogenesis. PMID- 15527761 TI - Genistein induces Gadd45 gene and G2/M cell cycle arrest in the DU145 human prostate cancer cell line. AB - Genistein is the most abundant isoflavone of soybeans and has been shown to cause growth arrest in various human cancer cell lines. However, the precise mechanism for this is still unclear. We report here that the growth arrest and DNA damage inducible gene 45 (gadd45) gene is induced by genistein via its promoter in a DU145 human prostate cancer cell line. The binding of transcription factor nuclear factor-Y to the CCAAT site of the gadd45 promoter appears to be important for this activation by genistein. PMID- 15527762 TI - Suppression of Sox6 in P19 cells leads to failure of neuronal differentiation by retinoic acid and induces retinoic acid-dependent apoptosis. AB - The Sox6 gene is a member of the Sox gene family, which encodes transcription factors, and previous studies have suggested that it plays an important role in the development of the central nervous system. Aggregation of embryonic carcinoma P19 cells with retinoic acid (RA) results in the development of neurons, glia, and fibroblast-like cells. Sox6 mRNA increases rapidly in P19 cells during RA induction and then decreases during differentiation into neuronal cells. To investigate whether Sox6 expression is essential for neuronal differentiation, we established Sox6-suppressed P19 (P19[anti-Sox6]) cells by transfection of antisense-Sox6 cDNA. Most of the P19[anti-Sox6] cells showed no neurites and were not stained by the anti-MAP 2 antibody, while the suppression of Sox6 expression nearly totally blocked neuronal differentiation in P19 cells. Further, Sox6 suppression caused RA-dependent apoptosis by P19[anti-Sox6] cells: RA-treated P19[anti-Sox6] cells showed chromatin condensation, DNA fragmentation, and an increase in caspase-3-like activity. Thus, Sox6 is considered essential for neuronal differentiation and may play an important role in the early stages of neuronal differentiation or apoptosis. PMID- 15527763 TI - Action of solamargine on human lung cancer cells--enhancement of the susceptibility of cancer cells to TNFs. AB - Solamargine (SM), isolated from Solanum incanum herb, displayed a superior cytotoxicity in four human lung cancer cell lines. The half-inhibitory concentrations (IC50), of the cell viability assay for H441, H520, H661 and H69 cells were 3, 6.7, 7.2 and 5.8 microM, respectively. SM-induced apoptosis of these cells by PS externalization in a dose-dependent manner and increased sub-G1 fraction were observed. Quenching of the expression of tumor necrosis factor receptors (TNFRs) during the progress of human lung carcinogenesis has been previously reported. SM may induce cell apoptosis via modulating the expression of TNFRs and their subsequent TRADD/FADD signal cascades. Subsequently, SM treatment increased the binding activities of TNF-alpha and TNF-beta to the lung cancers, and the intrinsic TNFs-resistant cancer cells became susceptible to TNF alpha and -beta. In addition, SM caused release of cytochrome c, downregulation of anti-apoptotic Bcl-2 and Bcl-xL, increase of caspase-3 activity, and DNA fragmentation. Thus, SM could modulate the expressions of TNFRs and Bcl-2, and might be a potential anticancer agent for TNFs and Bcl-2 related resistance of human lung cancer cells. PMID- 15527764 TI - Direct observation of the pH-dependent equilibrium between L-like and M intermediates of photoactive yellow protein. AB - Equilibrium between the photoproducts of photoactive yellow protein (PYP), present in a millisecond time scale, was studied. The near-UV intermediate of PYP (PYPM) was red-shifted by alkalization due to the deprotonation of the chromophore (pKa=10.2). In addition, a small amount of red-shifted intermediate coexisted with PYPM. Its spectral shape in the visible region agreed with that of PYPL, the precursor of PYPM. The fraction of PYPL-like product was maximal at pH 10. It decays with a rate constant identical to that of PYPM. These results indicate that PYPL-like product is in pH-dependent equilibrium with PYPM and deprotonated PYPM. PMID- 15527765 TI - Glycogen synthase kinase-3beta negatively regulates group IIA phospholipase A2 expression in human aortic smooth muscle and HepG2 hepatoma cells. AB - The present study shows that the IFN-gamma-mediated upregulation of secretory phospholipase A2 of group IIA (sPLA2-IIA) in HASMC and HepG2 cells is synergistically increased after simultaneous inhibition of glycogen synthase kinase-3beta (GSK-3beta) by indirubin-3'-monoxime, 5-iodo or AR-A014418. The effect of GSK-3beta inhibition was dose- and time-dependent and can be further augmented by its concomitant incubation with Clostridium difficile toxin B, an inhibitor of small Rho proteins, or H-1152, an inhibitor of Rho-associated kinase. Using AG-490 and caffeic acid phenethyl ester (CAPE), it is further demonstrated that the effect of GSK-3beta inhibition on sPLA2-IIA expression depends on Janus kinase-2 and NF-kappaB-signaling. PMID- 15527766 TI - Segregation of Nogo66 receptors into lipid rafts in rat brain and inhibition of Nogo66 signaling by cholesterol depletion. AB - NogoA, a myelin-associated component, inhibits neurite outgrowth. Nogo66, a portion of NogoA, binds to Nogo66 receptor (NgR) and induces the inhibitory signaling. LINGO-1 and p75 neurotrophin receptor (p75), the low-affinity nerve growth factor receptor, are also required for NogoA signaling. However, signaling mechanisms downstream to Nogo receptor remain poorly understood. Here, we observed that NgR and p75 were colocalized in low-density membrane raft fractions derived from forebrains and cerebella as well as from cerebellar granule cells. NgR interacted with p75 in lipid rafts. In addition, disruption of lipid rafts by beta-methylcyclodextrin, a cholesterol-binding reagent, reduced the Nogo66 signaling. Our results suggest an important role of lipid rafts in facilitating the interaction between NgRs and provide insight into mechanisms underlying the inhibition of neurite outgrowth by NogoA. PMID- 15527767 TI - Proteomics-based identification of proteins interacting with Smad3: SREBP-2 forms a complex with Smad3 and inhibits its transcriptional activity. AB - Smad3 is an important component of transforming growth factor-beta (TGFbeta) intracellular signalling. To identify novel interacting proteins of Smad3, we performed pull-down assays with Smad3 constructs fused to glutathione-S transferase. Proteins which formed complexes with these constructs were analyzed by two-dimensional gel electrophoresis, and were identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. We identified 14 proteins interacting with the Smad3 construct lacking the N-terminal Mad homology domain 1 (MH1), and 12 proteins interacting with the construct lacking the C terminal MH2 domain. Proteins involved in signalling processes, in metabolism regulation, novel proteins, and components of cytoskeleton form four groups of interacting proteins. Interactions of AGP7, sex-determining region Y protein, actin beta and sterol regulatory element binding protein-2 (SREBP-2) proteins with Smad3 constructs were confirmed by immunoblotting with specific antibodies. Interaction of Smad3 with SREBP-2 was also confirmed by co-immunoprecipitation of myc-Smad3 and Flag-SREBP-2 upon expression in mammalian cells. We found that SREBP-2 inhibited the transcriptional activity of Smad3 in luciferase reporter assays. PMID- 15527768 TI - The P3 domain of E. coli ribonuclease P RNA can be truncated and replaced. AB - We prepared some truncated and replaced P3 mutants of Escherichia coli RNase P RNA, and used them to examine the RNase P ribozyme and holoenzyme reactions of a pre-tRNA substrate. The results indicated that mutations in the P3 domain did not affect the cleavage site selection of the pre-tRNA substrate, but did affect the efficiency of cleavage of the substrate. Results of stepwise truncation of the P3 domain and its replacement by the TAR sequence showed that the P3 domain of the E. coli RNase P was able to be truncated to certain length and was replaceable, but could not be deleted in the ribozyme. PMID- 15527769 TI - Blue fluorescent protein from the calcium-sensitive photoprotein aequorin is a heat resistant enzyme, catalyzing the oxidation of coelenterazine. AB - Blue fluorescent protein from the calcium-sensitive photoprotein aequorin (BFP aq) was prepared and determined to be a heat resistant enzyme, catalyzing the luminescent oxidation of coelenterazine (luciferin) with molecular oxygen as a general luciferase. After treatment with excess ethylenediaminetetraacetic acid to remove Ca2+ from BFP-aq, the blue fluorescence shifted to a greenish fluorescence. This greenish fluorescent protein (gFP-aq) was identified as a non covalent complex of apoaequorin with coelenteramide (oxyluciferin) in a molar ratio of 1:1. By incubation with coelenterazine in the absence of reducing reagents, gFP-aq was converted to aequorin at 25 degrees C. BFP-aq and gFP-aq possessing both fluorescence and luminescence activities may work as novel reporter proteins. PMID- 15527770 TI - Crystal structure of human ISG20, an interferon-induced antiviral ribonuclease. AB - ISG20 is an interferon-induced antiviral exoribonuclease that acts on single stranded RNA and also has minor activity towards single-stranded DNA. It belongs to the DEDDh group of RNases of the DEDD exonuclease superfamily. We have solved the crystal structure of human ISG20 complexed with two Mn2+ ions and uridine 5' monophosphate (UMP) at 1.9 A resolution. Its structure, including that of the active site, is very similar to those of the corresponding domains of two DEDDh group DNases, the epsilon subunit of Escherichia coli DNA polymerase III and E. coli exonuclease I, strongly suggesting that its catalytic mechanism is identical to that of the two DNases. However, ISG20 also has distinctive residues, Met14 and Arg53, to accommodate hydrogen bonds with the 2'-OH group of the UMP ribose, and these residues may be responsible for the preference of ISG20 for RNA substrates. PMID- 15527771 TI - Islet amyloid polypeptide-induced membrane leakage involves uptake of lipids by forming amyloid fibers. AB - Fibril formation of islet amyloid polypeptide (IAPP) is associated with cell death of the insulin-producing pancreatic beta-cells in patients with Type 2 Diabetes Mellitus. A likely cause for the cytotoxicity of human IAPP is that it destroys the barrier properties of the cell membrane. Here, we show by fluorescence confocal microscopy on lipid vesicles that the process of hIAPP amyloid formation is accompanied by a loss of barrier function, whereby lipids are extracted from the membrane and taken up in the forming amyloid deposits. No membrane interaction was observed when preformed fibrils were used. It is proposed that lipid uptake from the cell membrane is responsible for amyloid induced membrane damage and that this represents a general mechanism underlying the cytotoxicity of amyloid forming proteins. PMID- 15527772 TI - The carboxy-terminus of the hepatitis B virus X protein is necessary and sufficient for the activation of hypoxia-inducible factor-1alpha. AB - Hepatitis B virus X protein (HBx) of the hepatitis B virus is strongly implicated in angiogenesis and metastasis during hepatocarcinogenesis. Previously, we reported that HBx enhances activity of hypoxia-inducible factor-1alpha (HIF 1alpha), a potent transactivator that induces angiogenic factors. Here, we delineate the structural region of HBx that potentiates HIF-1alpha. The carboxy terminus of HBx increased the stability of HIF-1alpha protein, probably through inhibiting interaction with von Hippel-Lindau protein. Further, the carboxy terminus of HBx enhanced the transactivation function of HIF-1alpha by enhancing its association with CREB binding protein (CBP). Finally, we demonstrated the physical association of HBx with the basic helix-loop-helix/PER-ARNT-SIM domain, the inhibitory domain, and the carboxy-terminal transactivation domain of HIF 1alpha in vivo. PMID- 15527773 TI - Amyloid aggregates of the prion peptide PrP106-126 are destabilised by oxidation and by the action of dendrimers. AB - The prion protein (PrP) peptide 106-126 forms amyloid aggregates in vitro and this sequence is speculated to be involved in the formation of amyloid fibrils by the abnormally folded PrP protein (PrPSc) found in spongiform encephalopathies. It is shown here by incubation experiments in water using Thioflavin T (ThT) as a fluorescent probe for amyloid formation that changes in C-terminal charge, oxidation state and conformational stabilisation lead to large changes in amyloid forming behaviour (amyloidogenicity) of this peptide. Amyloid formation is favoured by a charged C-terminus and is strongly inhibited by oxidation. Furthermore, cationic dendrimers are shown to perturb peptide fibrillation in a process dependent on the nature of the charged groups on the dendrimer surface. PMID- 15527774 TI - Cooperative binding of the hnRNP K three KH domains to mRNA targets. AB - The heterogeneous nuclear ribonucleoprotein (hnRNP) K homology (KH) domain is an evolutionarily conserved module that binds short ribonucleotide sequences. KH domains most often are present in multiple copies per protein. In vitro studies of hnRNP K and other KH domain bearing proteins have yielded conflicting results regarding the relative contribution of each KH domain to the binding of target RNAs. To assess this RNA-binding we used full-length hnRNP K, its fragments and the yeast ortholog as baits in the yeast three-hybrid system. The results demonstrate that in this heterologous in vivo system, the three KH domains bind RNA synergistically and that a single KH domain, in comparison, binds RNA weakly. PMID- 15527775 TI - Identification of a novel shrimp protein phosphatase and its association with latency-related ORF427 of white spot syndrome virus. AB - To characterize the role of latency-associated ORF427 of white spot syndrome virus (WSSV), a shrimp cDNA library was constructed to screen interacting proteins of ORF427. Employing the yeast two-hybrid system, a novel shrimp protein phosphatase (named PPs), sharing 93% homology with human protein phosphatase 1, has been identified able to bind ORF427 in yeast. Through co-immunoprecipitation assays, the interaction between PPs and ORF427 was further confirmed both in vitro and in vivo. Interestingly, the novel shrimp protein phosphatase consists of only 199 aa and contains almost all the functional catalytic domains of human protein phosphatase, while it lacks the corresponding C-terminal non-catalytic sequence. Transcription and translation products of the identified cDNA can be detected in both normal and WSSV-infected shrimps; and PPs was found to localize mainly in the lysosome of shrimp cells. To characterize its function, the PPs cDNA was highly expressed in bacteria and the purified protein showed phosphatase activity when tested against pNPP in a standard phosphatase assay. Our results suggest that the identified protein phosphatase, PPs, may represent a novel member of protein phosphatase family and might be involved in the regulation of WSSV's life cycle through interaction with latency-related ORF427 of WSSV. PMID- 15527776 TI - Signal transduction mechanism of the seabream growth hormone secretagogue receptor. AB - We have recently cloned the full-length cDNAs of the two growth hormone secretagogue receptor (GHSR) subtypes from a teleost species, the black seabream (Acanthopagrus schlegeli) [Mol. Cell. Endocrinol. 214 (2004) 81], namely sbGHSR 1a and sbGHSR-1b. Functional expression of these two receptor constructs in human embryonic kidney 293 (HEK293) cells indicated that stimulation of sbGHSR-1a by growth hormone secretagogues (GHS) could evoke increases in intracellular Ca2+ concentration ([Ca2+]i), whereas sbGHSR-1b appeared to play an inhibitory role on the signal transduction activity of sbGHSR-1a. In the present study, we have further investigated the signal transduction mechanism of sbGHSR-1a. The peptide GHS GHRP-6 and the non-peptide GHS L163,540 were able to trigger a receptor specific and phospholipase C (PLC)-dependent elevation of [Ca2+]i in HEK293 cells stably expressing sbGHSR-1a. This GHS-induced calcium mobilization was also dependent on protein kinase C activated L-type calcium channel opening. It was found that sbGHSR-1a could function in an agonist-independent manner as it exhibited a high basal activity of inositol phosphate production in the absence of GHS, indicating that the fish receptor is constitutively active. In addition, the extracellular signal-regulated kinases 1 and 2 (ERK1/2) were found to be activated upon stimulation of sbGHSR-1a by GHRP-6. This observation provides direct evidence in the coupling of sbGHSR-1a to ERK1/2 activation. Neither Gs nor Gi proteins are coupled to the receptor, as GHS did not induce cAMP production nor inhibit forskolin-stimulated cAMP accumulation in the sbGHSR-1a bearing cells. Furthermore, the ability of the GHSR antagonist D-Lys3-GHRP-6 to inhibit basal PLC and basal ERK1/2 activity suggests that this compound is an inverse agonist. In summary, the sbGHSR-1a appears to couple through the G(q/11)-mediated pathway to activate PLC, resulting in increased IP3 production and Ca2+ mobilization from both intracellular and extracellular stores. Moreover, sbGHSR 1a may trigger multiple signal transduction cascades to exert its physiological functions. PMID- 15527777 TI - Subcellular localization of liver FBPase is modulated by metabolic conditions. AB - In primary cultured hepatocytes, fructose-1,6-bisphosphatase (FBPase) localization is modulated by glucose, dihydroxyacetone (DHA) and insulin. In the absence of these substrates, FBPase was present in the cytoplasm, but the addition of glucose or DHA induced its translocation to the nucleus. As expected, we observed the opposite effect of glucose on glucokinase localization. The addition of insulin in the absence of glucose largely increased the amount of nuclear FBPase. Moreover, at high concentrations of glucose or DHA, FBPase shifted from the cytosol to the cell periphery and co-localized with GS. Interestingly, the synthesis of Glu-6-P and glycogen induced by DHA was not inhibited by insulin. These results indicate that FBPase is involved in glycogen synthesis from gluconeogenic precursors. Overall, these findings show that translocation may be a new integrative mechanism for gluconeogenesis and glyconeogenesis. PMID- 15527778 TI - Evolutional insights on uncharacterized SARS coronavirus genes. AB - The complete genome of the severe acute respiratory syndrome coronavirus (SARS CoV) and many of its variants has been determined by several laboratories. The genome contains fourteen predicted open reading frames (ORFs). However, a function had been clearly assigned for only six of these ORFs, in the viral replication, transcription and structural constituents. The others are herein referred to as uncharacterized ORFs (UC-ORFs). Here, we try to provide a relational insight on those UC-ORFs, suggesting that a number of them are remotely related to structural proteins of coronaviruses and other viruses infecting mammalian hosts. Surprisingly, several of the UC-ORFs exhibit considerable similarity with other SARS-CoV ORFs. These observations may provide clues on the evolution and genome dynamics of the SARS-CoV. PMID- 15527779 TI - Inhibition of furin by serpin Spn4A from Drosophila melanogaster. AB - The serpin gene Spn4 from Drosophila melanogaster encodes multiple isoforms with alternative reactive site loops (RSL). Here, we show that isoform Spn4A inhibits human furin with an apparent kassoc of 5.5 x 10(6) M(-1) s(-1). The serpin forms SDS-stable complexes with the enzyme and the RSL of Spn4A is cleaved C-terminally to the sequence -Arg-Arg-Lys-Arg/ in accord with the recognition/cleavage site of furin. Immunofluorescence studies show that Spn4A is localized in the endoplasmic reticulum (ER), suggesting that the inhibitor is an interesting tool for investigating the cellular mechanisms regulating furin and for the design of agents controlling prohormone convertases. PMID- 15527780 TI - Structure of the mitochondrial beta-ketoacyl-[acyl carrier protein] synthase from Arabidopsis and its role in fatty acid synthesis. AB - Mitochondrial fatty acid synthesis is catalyzed by a dissociated fatty acid synthase similar to those of plant plastids and bacteria. The crystal structure of a mitochondrial beta-ketoacyl-[acyl carrier protein] synthase (mtKAS), namely that from Arabidopsis thaliana, has been determined for the first time. This enzyme accomplishes the vital condensation steps in constructing fatty acid carbon skeletons. The product profile of mtKAS is unusual in that C8 and C(14-16) fatty acyl chains predominate. An enzyme architecture that likely is the basis for the observed bimodal profile of mtKAS products can be derived from the shape of the acyl binding pocket. PMID- 15527781 TI - Double transfection improves small interfering RNA-induced thrombin receptor (PAR 1) gene silencing in DU 145 prostate cancer cells. AB - The efficiency of small interfering RNA (siRNA)-induced gene knockdown is hampered by low transfection efficiency. We established a novel and simple double transfection method using specific siRNA duplexes targeted against human thrombin receptor PAR-1 in DU 145 prostate cancer cells. The initial siRNA transfection of cell suspensions followed by re-transfection of adherent cells on the following day resulted in undetectable PAR-1 mRNA and absent receptor protein. PAR-1 mRNA expression was silenced for up to five days. Functional studies showed that PAR-1 gene silencing in DU 145 cells abolished the modulating effects of thrombin on cell adhesion to the extracellular matrix proteins, fibronectin and laminin, thus demonstrating the essential role of PAR-1 in mediating thrombin effects on DU 145 cell adhesion. PMID- 15527782 TI - A new type of protein methylation activated by tyrphostin A25 and vanadate. AB - It has been reported that S-adenosylmethionine-dependent protein methylation in rat kidney extracts can be greatly stimulated by tyrphostin A25, a tyrosine kinase inhibitor. We have investigated the nature of this stimulation. We find that addition of tyrphostin A25, in combination with the protein phosphatase inhibitor vanadate, leads to the stimulation of methylation of polypeptides of 64, 42, 40, 36, 31, and 15 kDa in cytosolic extracts of mouse kidney. The effect of tyrphostin appears to be relatively specific for the A25 species. The enhanced methylation does not represent the activity of the families of protein histidine, lysine or arginine methyltransferases, nor that of the l-isoaspartyl/d-aspartyl methyltransferase, enzymes responsible for the bulk of protein methylation in most cell types. Chemical and enzymatic analyses of the methylated polypeptides suggest that the methyl group is in an ester linkage to the protein. In heart extracts, we find a similar situation but here the stimulation of methylation is not dependent upon vanadate and an additional 18 kDa methylated species is found. In contrast, little or no stimulation of methylation is found in brain or testis extracts. This work provides evidence for a novel type of protein carboxyl methylation reaction that may play a role in signaling reactions in certain mammalian tissues. PMID- 15527783 TI - Tyrosine dephosphorylation of STAT3 in SARS coronavirus-infected Vero E6 cells. AB - Severe acute respiratory syndrome (SARS) has become a global public health emergency. p38 mitogen-activated protein kinase (MAPK) and its downstream targets are activated in SARS coronavirus (SARS-CoV)-infected Vero E6 cells and activation of p38 MAPK enhances the cytopathic effects of SARS-CoV infection. In addition, weak activation of Akt cannot prevent SARS-CoV infection-induced apoptosis in Vero E6 cells. In the present study, we demonstrated that signal transducer and activator of transcription (STAT) 3, which is constitutively phosphorylated at tyrosine (Tyr)-705 and slightly phosphorylated at serine (Ser) 727 in Vero E6 cells, was dephosphorylated at Tyr-705 on SARS-CoV infection. In addition to phosphorylation of p38 MAPK in virus-infected cells, other MAPKs, i.e., extracellular signal-regulated kinase (ERK) 1/2 and c-Jun N-terminal kinase (JNK), were phosphorylated. Although inhibitors of ERK1/2 and JNK (PD98059 and SP600125) had no effect on phosphorylation status of STAT3, inhibitors of p38 MAPK (SB203580 and SB202190) partially inhibited dephosphorylation of STAT3 at Tyr-705. Tyr-705-phosphorylated STAT3 was localized mainly in the nucleus in mock infected cells, whereas STAT3 disappeared from the nucleus in virus-infected cells. As STAT3 acts as an activator of transcription in the nucleus, these results suggest that STAT3 lacks its activity on transcription in SARS-CoV infected Vero E6 cells. PMID- 15527784 TI - Identification of an Arabidopsis thaliana gene for cardiolipin synthase located in mitochondria. AB - Cardiolipin (CL) is an anionic phospholipid with a dimeric structure. In eukaryotes, it is primarily localized in the inner membranes of mitochondria. Although the biosynthetic pathway of CL is well known, the gene for CL synthase has not been identified in any higher organisms. In this study, the CLS gene for a CL synthase has been identified in a higher plant, Arabidopsis thaliana. We have shown that the CLS gene encodes a CL synthase by demonstrating its ability to catalyze the reaction of CL synthesis from CDP-diacylglycerol and phosphatidylglycerol, and that CLS is targeted into mitochondria. These findings demonstrate that CLS is a CL synthase located in mitochondria. PMID- 15527785 TI - Consumption of nitric oxide by endothelial cells: evidence for the involvement of a NAD(P)H-, flavin- and heme-dependent dioxygenase reaction. AB - In the present study, we investigated the mechanism of nitric oxide (NO) inactivation by endothelial cells. All experiments were performed in the presence of superoxide dismutase to minimize the peroxynitrite reaction. Incubation of the NO donor diethylamine/NO adduct with increasing amounts of intact cells led to a progressive decrease of the NO concentration, demonstrating a cell-dependent consumption of NO. In cell homogenates, consumption of NO critically depended on the presence of NADPH or NADH and resulted in the formation of nitrate. Both NO consumption and nitrate formation were largely inhibited by the heme poisons NaCN and phenylhydrazine as well as the flavoenzyme inhibitor diphenylene iodonium. Further characterization of this NO consumption pathway suggests that endothelial cells express a unique membrane-associated enzyme or enzyme system analogous to the bacterial NO dioxygenase that converts NO to nitrate in a NAD(P)H-, flavin- and heme-dependent manner. PMID- 15527786 TI - The importance of Loop 7 for the activity of calcineurin. AB - Calcineurin (CN) is a heterodimer composed of a catalytic subunit (CNA) and a regulatory subunit (CNB). Loop 7 lies within the CNA catalytic domain. To investigate the role of Loop 7 in enzyme activity, we systematically examined all its residues by site-directed deletion mutation. Our results show that the Loop 7 residues are important for enzyme activity. Besides deleting residues V314, Y315 or N316, enzyme activity also increased dramatically when residues D313 or K318 were deleted. In contrast, almost all activity was lost when L312 or N317 were deleted. Ni2+ and Mn2+ were effective activators for all active mutants. However, whereas the wild-type enzyme was more efficiently activated by Ni2+ than by Mn2+ with 32P-labeled R(II) peptide as substrate, the reverse was true in all the mutants. We also found that the effect of Loop 7 on enzyme activity was substrate dependent, and involved interactions between Loop 7 residues and the unresolved part of the CN crystal structure near the auto-inhibitory domain and catalytic site. PMID- 15527787 TI - Purification and primary structure of two isoforms of arenicin, a novel antimicrobial peptide from marine polychaeta Arenicola marina. AB - Two novel 21-residue antimicrobial peptides, arenicin-1 and arenicin-2, exhibiting activity against Gram-positive and Gram-negative bacteria and fungi, were purified from coelomocytes of marine polychaeta Arenicola marina (lugworm) by preparative gel electrophoresis and RP-HPLC. Molecular masses (2758.3 and 2772.3 Da) and complete amino acid sequences (RWCVYAYVRVRGVLVRYRRCW and RWCVYAYVRIRGVLVRYRRCW) were determined for each isoform. Each arenicin has one disulfide bond (Cys3-Cys20). The total RNA was isolated from the lugworm coelomocytes, RT-PCR and cloning were performed, and cDNA was sequenced. A 202 residue preproarenicin contains a putative signal peptide (25 amino acids) and a long prodomain. Arenicins have no structure similarity to any previously identified antimicrobial peptides. PMID- 15527788 TI - The HECT E3 ubiquitin ligase Rsp5 is important for ubiquitin homeostasis in yeast. AB - The HECT E3 ubiquitin ligase Rsp5, a yeast member of the Nedd4 family, has been implicated in many different aspects of cell physiology. Here, we present evidence that Rsp5 function is important for ubiquitin homeostasis. Several observations suggest that ubiquitin is limiting in the rsp5-1 mutant. Reduced synthesis of ubiquitin appears to contribute to ubiquitin depletion. A transient inhibition of general protein synthesis is observed in a wildtype strain upon heat-shock. While the wildtype cells quickly recover from this transient arrest, the rsp5-1 cells remain arrested. This suggests that Rsp5 is important for recovery from heat-induced protein synthesis arrest. Our results suggest that rsp5 phenotypes should be interpreted with caution, since some of the phenotypes could be simply the result of ubiquitin limitation. PMID- 15527789 TI - Mitogen-activated protein kinase phosphatase-1 (MKP-1): >100-fold nocturnal and norepinephrine-induced changes in the rat pineal gland. AB - The norepinephrine-driven increase in mitogen-activated protein kinase (MAPK) activity is part of the mechanism that regulates arylalkylamine N acetyltransferase (AA-NAT) activity in the rat pineal gland. We now report a marked nocturnal increase in the expression of a MAPK phosphatase, MAP kinase phosphatase-1 (MKP-1), that was blocked by maintaining animals in constant light or treatment with propranolol. MKP-1 expression was regulated by norepinephrine acting through both alpha- and beta-adrenergic receptors. These results establish a nocturnal increase in pineal MKP-1 expression that is under the control of a photoneural system. Because substrates of MKP-1 can influence AA-NAT activity, our findings suggest the involvement of MKP-1 in the regulation of the nocturnal AA-NAT signal. PMID- 15527790 TI - A brilliant monomeric red fluorescent protein to visualize cytoskeleton dynamics in Dictyostelium. AB - Red fluorescent proteins (RFPs) combined with GFP are attractive probes for double-fluorescence labeling of proteins in live cells. However, the application of these proteins is restrained by stable oligomer formation and by their weak fluorescence in vivo. Previous attempts to eliminate these problems by mutagenesis of RFP from Discosoma (DsRed) resulted in the monomeric mRFP1 and in the tetrameric RedStar RFP, which is distinguished by its enhanced fluorescence in vivo. Based on these mutations, we have generated an enhanced monomeric RFP, mRFPmars, and report its spectral properties. Together with green fluorescent labels, we used mRFPmars to visualize filamentous actin structures and microtubules in Dictyostelium cells. This enhanced RFP proved to be suitable to monitor the dynamics of cytoskeletal proteins in cell motility, mitosis, and endocytosis using dual-wavelength fluorescence microscopy. PMID- 15527791 TI - Allele-specific transcript isoforms in human. AB - Estimates of the number of human genes that produce more than one transcript isoform through alternative mRNA splicing depend on the assumption that the observation of multiple transcripts from a gene can be attributed entirely to alternative splicing. It is possible, however, that a substantial proportion of cases where multiple transcripts have been observed for a gene result from differences between alleles. Many examples of genes that are spliced differently from different alleles have been reported but no systematic estimate of the proportion of alternatively spliced genes that are affected by such polymorphisms has been carried out. We find that alternative transcript isoforms are non randomly associated with closely linked nucleotide polymorphisms, based on an integrated analysis of the dbSNP, dbEST and ASAP databases. From the observed level of association between transcript isoforms and polymorphisms, we estimate that 21% of alternatively spliced genes are affected by polymorphisms that either completely determine which form of the transcript is observed or alter the relative abundances of some of the alternative isoforms. We provide a conservative lower bound of 6% on this estimate and point out that alternative splicing cannot be confirmed absolutely unless more than one transcript is observed from the same allele. PMID- 15527792 TI - Potential anti-atherogenic cell action of the naturally occurring 4-O-methyl derivative of gallic acid on Ang II-treated macrophages. AB - We have recently established that the blood concentrations of gallic acid (GA), a polyphenolic component naturally found in food, and its O-methyl derivatives are very low (practically < or = 1 microM) in physiological (postprandial) condition. Using acellular oxidant systems and macrophage-differentiated promonocytes (MDPs) THP-1, we show here that the direct and indirect (through depressing effect on the superoxide cell production) antioxidant properties of these components were not effective at these concentrations. In contrast, 4-O-methyl GA was the most efficient component to depress AT1R and CD36 mRNA expression in Ang II-treated MDPs, suggesting a strong inhibition of Ang II-triggered pro-atherogenic mechanisms of foam cell formation. PMID- 15527793 TI - The poison Dart frog's batrachotoxin modulates Nav1.8. AB - Batrachotoxin is a potent modulator of voltage-gated sodium channels, leading to irreversible depolarisation of nerves and muscles, fibrillation, arrhythmias and eventually cardiac failure. Since its discovery, field researchers also reported numbness after their skin came into contact with this toxin. Intrigued by this phenomenon, we determined the effect of batrachotoxin on the voltage-gated sodium channel Nav1.8, which is considered to be a key player in nociception. As a result, we discovered that batrachotoxin profoundly modulates this channel: the inactivation process is severely altered, the voltage-dependence of activation is shifted towards more hyperpolarised potentials resulting in the opening of Nav1.8 at more negative membrane potentials and the ion selectivity is modified. PMID- 15527794 TI - Adrenomedullin signals through NF-kappaB in epithelial cells. AB - Adrenomedullin is a peptide found in a variety of cells and tissues and involved in a multitude of biological processes. Recently, adrenomedullin has been identified as a host defense peptide and as such it plays a role in the inflammatory response. The transcription factor NF-kappaB is a major regulator of genes involved in the inflammatory response and the aim of this study was to determine whether NF-kappaB played a role in the inflammatory process triggered by adrenomedullin. Skin epithelial cells (HaCaTs) were used as our model in vitro. Western blot analysis from adrenomedullin-stimulated HaCaT cells revealed a rapid degradation of NF-kappaB inhibitor alpha and beta followed by the translocation of free NF-kappaB to the nucleus, where it was detected by Texas Red immunostaining after incubation with adrenomedullin for 15 min. Electromobility shift assay showed that NF-kappaB present in the nucleus was active, since it bound to a probe containing an NF-kappaB binding site. Supershift assays indicated that p50 and p65, members of the NF-kappaB family, were both part of the NF-kappaB dimmers involved in adrenomedullin cell signaling. HaCaTs secreted interleukin-6 in response to AM, which was significantly attenuated by the NF-kappaB inhibitor SN-50. Taken together, the data lend support for an immunoregulatory role for AM. PMID- 15527795 TI - Biochemical characterization of the major adenylyl cyclase, Cya1, in the cyanobacterium Synechocystis sp. PCC 6803. AB - We report herein the biochemical properties of an adenylyl cyclase, Cya1, from the cyanobacterium Synechocystis sp. PCC 6803. Heterologously expressed Cya1 catalyzed cyclic AMP formation with a Km for ATP of approximately 2.2 microM at pH 7.5. Although cellular Cya1 activity is increased by blue light illumination [Terauchi and Ohmori, Mol. Microbiol. 52 (2004) 303], purified Cya1 did not contain any chromophores, and the activity was light-insensitive. This suggests that an unknown blue light-responsive factor interacts with the N-terminal regulatory domain of Cya1 to control its adenylyl cyclase activity. Finally, our results show that the sensor of blue light using FAD (BLUF) protein, Slr1694, does not appear to be involved in the regulation of Cya1-mediated cAMP signal transduction in this bacterium. PMID- 15527796 TI - Conserved transcriptional regulation of a cone phototransduction gene in vertebrates. AB - cGMP-phosphodiesterase (PDE) is a key component in visual phototransduction. Rod and cone photoreceptors each produce their unique cGMP-PDE subunits. The alpha' catalytic subunits are believed to be cone-specific. In this study, we report that transfection of the -132 to +139 sequence in the upstream region of the human alpha'-PDE gene fused to luciferase cDNA gives the highest level of reporter gene transcription in cultured retinoblastoma Y79 cells. Transgenic Xenopus laevis carrying this sequence fused to green fluorescent protein (GFP) expressed GFP in cones, suggesting a conserved regulatory mechanism for alpha' PDE transcription in both human and frog. PMID- 15527797 TI - Contribution of the active site aspartic acid to catalysis in the bacterial neuraminidase from Micromonospora viridifaciens. AB - A recombinant D92G mutant sialidase from Micromonospora viridifaciens has been cloned, expressed and purified. Kinetic studies reveal that the replacement of the conserved aspartic acid with glycine results in a catalytically competent retaining sialidase that possesses significant activity against activated substrates. The contribution of this aspartate residue to the free energy of hydrolysis for natural substrates is greater than 19 kJ/mol. The three dimensional structure of the D92G mutant shows that the removal of aspartic acid 92 causes no significant re-arrangement of the active site, and that an ordered water molecule substitutes for the carboxylate group of D92. PMID- 15527798 TI - A novel site of AKT-mediated phosphorylation in the human MDM2 onco-protein. AB - MDM2 is an E3 ubiquitin ligase which mediates ubiquitylation and proteasome dependent degradation of the p53 tumor suppressor protein. Phosphorylation of MDM2 by the protein kinase AKT is thought to regulate MDM2 function in response to survival signals, but there has been uncertainty concerning the identity of the sites phosphorylated by AKT. In the present study, we identify Ser-166, a site previously reported as an AKT target, and Ser-188, a novel site which is the major site of phosphorylation of MDM2 by AKT in vitro. Analysis of MDM2 in cultured cells confirms that Ser-166 and Ser-188 are phosphorylated by AKT in a physiological context. PMID- 15527799 TI - Basic fibroblast growth factor enhances PPARgamma ligand-induced adipogenesis of mesenchymal stem cells. AB - Mesenchymal stem cells (MSCs) are capable of differentiating into a variety of lineages, including bone, cartilage, or fat, depending on the inducing stimuli and specific growth and differentiation factors. It is widely acknowledged that basic fibroblast growth factor (bFGF) modulates chondrogenic and osteogenic differentiation of MSCs, but thorough investigations of its effects on adipogenic differentiation are lacking. In this study, we demonstrate on the cellular and molecular level that supplementation of bFGF in different phases of cell culture leads to a strong enhancement of adipogenesis of MSCs, as induced by an adipogenic hormonal cocktail. In cultures receiving bFGF, mRNA expression of peroxisome proliferator-activated receptor gamma2 (PPARgamma2), a key transcription factor in adipogenesis, was upregulated even prior to adipogenic induction. In order to investigate the effects of bFGF on PPARgamma ligand induced adipogenic differentiation, the thiazolidinedione troglitazone was administered as a single adipogenic inducer. Basic FGF was demonstrated to also strongly increase adipogenesis induced by troglitazone, that is, bFGF clearly increased the responsiveness of MSCs to a PPARgamma ligand. PMID- 15527800 TI - Interaction of calmodulin with the phosphofructokinase target sequence. AB - Ca4.calmodulin (Ca4.CaM) inhibits the glycolytic enzyme phosphofructokinase, by preventing formation of its active tetramer. Fluorescence titrations show that the affinity of complex formation of Ca4.CaM with the key 21-residue target peptide increases 1000-fold from pH 9.0 to 4.8, suggesting the involvement of histidine and carboxylic acid residues. 1H NMR pH titration indicates a marked increase in pKa of the peptide histidine on complex formation and HSQC spectra show related pH-dependent changes in the conformation of the complex. This unusually strong sensitivity of a CaM-target complex to pH suggests a potential functional role for Ca4.CaM in regulation of the glycolytic pathway. PMID- 15527801 TI - Amino-terminal domain of ATRIP contributes to intranuclear relocation of the ATR ATRIP complex following DNA damage. AB - ATM and rad3-related protein kinase (ATR), a member of the phosphoinositide kinase-like protein kinase family, plays a critical role in cellular responses to DNA structural abnormalities in conjunction with its interacting protein, ATRIP. Here, we show that the amino-terminal portion of ATRIP is relocalized to DNA damage-induced nuclear foci in an RPA-dependent manner, despite its lack of ability to associate with ATR. In addition, ATR-free ATRIP protein can be recruited to the nuclear foci. Our results suggest that the N-terminal domain of the ATRIP protein contributes to the cell cycle checkpoint by regulating the intranuclear localization of ATR. PMID- 15527802 TI - The salt bridge of calcineurin is important for transferring the effect of CNB binding to CNA. AB - Calcineurin (CN) is a heterodimer consisting of a catalytic subunit (CNA) and a regulatory subunit (CNB). The crystal structure shows that three residues or regions of CNA are mainly responsible for the interaction with CNB: the CNB binding helix (BBH), the N-terminus, and Glu53 that forms a salt bridge with Lys134 of CNB. In this report, we try to find the role that the salt bridge plays in the interaction between CNA and CNB. We found that mutation of Glu53 greatly reduced its responsiveness to CNB in the phosphatase assay and also that mutation of Lys134 of CNB affected its ability to activate the phosphatase activity of CNA. Structural analysis showed that disruption of the salt bridge affected the compact association of CNA and CNB. Thus, the salt bridge appears to help to stabilize CN and transfer the effects of CNB binding to CNA to activate its phosphatase activity. PMID- 15527804 TI - Photosystem II associated carbonic anhydrase activity in higher plants is situated in core complex. AB - The thylakoid membrane containing photosystem II (PSII membranes) from pea and wheat leaves catalyzed the reaction of CO2 hydration with low rate, which increased after their incubation either with Triton X-100, up to Triton/chlorophyll ratio 1:1, or 1 M CaCl2. The presence of the inhibitor of CAs, p-aminomethylbenzensulfonamide (mafenide), at the start line in the course of electrophoresis of PSII membranes solubilized by n-dodecyl-beta-maltoside (DM) decreased the amount of PSII core complex in the gel. The elution of PSII core complex from the column with immobilized mafenide occurred only either by mafenide or another inhibitor of CAs, ethoxyzolamide. The above results led to a conclusion that membrane-bound CA activity associated with PSII is situated in the core complex. PMID- 15527803 TI - Second transmembrane domain of human uncoupling protein 2 is essential for its anion channel formation. AB - Uncoupling proteins (UCP) are known to transport anions, such as Cl-, in addition to H+ transport. Although H+ transport by UCP is clearly involved in thermogenesis, the mechanism of its anion transport is not clearly understood. In this study, we examined the anion channel characteristics of the six individual helical transmembrane (TM) domains of the human UCP2. The second TM domain peptide (TM2) forms multi-state channels by assemblies of conductive oligomers. Furthermore, the TM2 exhibited voltage-dependent anion channels with properties comparable to those of UCP1 chloride channel. However, the other five TM peptides did not form UCP1-like channels. Moreover, an analog of TM2 in which two Arg residues were substituted by Ala residues did not form stable channels, implying the significance of Arg residues for anion transport. These results suggest that the anion channel structure of UCP2 protein is oligomeric and the second TM domain is essential for the voltage-dependence of this anion channel. PMID- 15527805 TI - Ethanol acts as a potent agent sensitizing colon cancer cells to the TRAIL induced apoptosis. AB - Identification of mechanisms of modulation of the TNF-related apoptosis-inducing ligand (TRAIL)-induced apoptosis is important for its potential use in anticancer therapy. Ethanol can induce cell death in vitro and in vivo by different signalling pathways. Its effect in combination with death ligands is unknown. We investigated how ethanol modulates the effects of TRAIL in colon cancer cells. After combined TRAIL and ethanol treatment, a potentiation of caspase-8, -9, -3 activation, a proapoptotic Bid protein cleavage, a decrease of mitochondrial membrane potential, a complete poly(ADP)ribose polymerase cleavage, and disappearance of antiapoptotic Mcl-1 protein were demonstrated. Ethanol acts as a potent agent sensitizing colon cancer cells to TRAIL-induced apoptosis. PMID- 15527807 TI - The pathophysiology, clinical presentation, and diagnosis of colon cancer and adenomatous polyps. AB - A review of the pathophysiology, clinical presentation, and diagnosis of colon cancer and colonic polyps is important and timely. This field is rapidly changing because of breakthroughs in the molecular basis of carcinogenesis and in the technology for colon cancer detection and treatment. This article reviews colon cancer and colonic polyps, with a focus on recent dramatic advances, to help the pri-mary care physician and internist appropriately refer patients for screening colonoscopy and intelligently evaluate colonoscopic findings to reduce the mortality from this cancer. PMID- 15527808 TI - Screening of patients at average risk for colon cancer. AB - Both the incidence and the mortality from colorectal cancer can be substantially reduced by primary and secondary prevention. There are many screening tests for colorectal cancer, and any test should result in a reduction in colorectal cancer incidence and mortality. If the age-eligible population undergoes these screening tests, the burden of colorectal cancer should be substantially reduced. The scientific evidence related to secondary prevention, specifically screening of individuals at average risk for colorectal cancer, is presently reviewed. PMID- 15527809 TI - Surveillance of patients at high risk for colorectal cancer. AB - Colorectal cancer (CRC) mortality may be greatly reduced by clinically feasible screening programs. The benefits of surveillance of high-risk programs are evident. Cancer mortality can be dramatically reduced by eradication of precursor lesions and by detection of cancer at an early and highly curable stage. Available screening methods, recommended intervals, and screening for other associated cancers are reviewed for specific high-risk groups. PMID- 15527810 TI - Prevention and therapy of colorectal cancer. AB - Colorectal cancer is expected to affect more than 146,000 and kill more than 57,000 Americans in 2004. Increased understanding of carcinogenesis is transforming clinical approaches to all stages of this disease. During the last 5 years, four new drugs have been approved for colorectal cancer treatment, and substantial progress has been made in identifying and developing agents that prevent or delay carcinogenesis. These advances substantiate target-driven approaches to cancer prevention and treatment, and provide fruitful opportunities for future investigations. PMID- 15527811 TI - The emerging role of virtual colonoscopy. AB - A review of the current status of virtual colonoscopy is germane and topical. Clinicians need to be knowledgeable about this rapidly evolving and clinically relevant technology to understand the test benefits and limitations and to refer patients for this test appropriately. A critical review of the exponentially expanding literature on this subject is important. This article describes the imaging meth-ods, adjunctive techniques, and radiologic interpretation of CT colonography, and comprehensively and critically reviews the clinical data to help the clinician evaluate the current and potential applications of this technology. PMID- 15527812 TI - Endoscopic ultrasound in the diagnosis, staging, and management of gastrointestinal and adjacent malignancies. AB - Endoscopic ultrasound (EUS) is a superior modality for local staging of gastrointestinal cancer. In interventional endosonography linear array echoendoscopes permit real-time EUS-guided puncture of target lesions for cytologic evaluation of such lesions. This article describes the basic principles of EUS, established indications pertaining to gastrointestinal cancer and other malignancies, and emerging indications for this minimally invasive technology. PMID- 15527813 TI - Novel endoscopic therapies for gastrointestinal malignancies: endoscopic mucosal resection and endoscopic ablation. AB - Gastrointestinal malignancies are often detected at advanced stages when the prognosis is poor. Screening guidelines that vary accord-ing to the regional disease prevalence are needed. High-resolution endoscopy, magnification endoscopy, chromoendoscopy, light autofluorescence endoscopy, and optical coherence tomography are new technologies designed to improve endoscopic detection. Once detected, lesions must be accurately staged, including depth of mucosal penetration and lymph node involvement, to determine endoscopic resectability. Widely applicable, relatively safe, and minimally invasive alternatives to surgery are needed. Endoscopic mucosal resection and endoscopic ablation are potentially curative for malignancies limited to the mucosa, obviating the need for surgery in these patients. PMID- 15527814 TI - Laparoscopic surgery for the prevention, palliation, and cure of gastrointestinal malignancies. AB - The development of surgical laparoscopic techniques has revolutionized the way surgeons approach many diseases, including cancer. This article briefly discusses the historical development of surgical laparoscopy; describes laparoscopic surgical techniques, with a focus on techniques for common intra-abdominal malignancies; and reviews laparoscopic management of common gastrointestinal malignancies. PMID- 15527815 TI - The MinK-related peptides. AB - Voltage-gated potassium (Kv) channels mediate rapid, selective diffusion of K+ ions through the plasma membrane, controlling cell excitability, secretion and signal transduction. KCNE genes encode a family of single transmembrane domain proteins called MinK-related peptides (MiRPs) that function as ancillary or beta subunits of Kv channels. When co-expressed in heterologous systems, MiRPs confer changes in Kv channel conductance, gating kinetics and pharmacology, and are fundamental to recapitulation of the properties of some native currents. Inherited mutations in KCNE genes are associated with diseases of cardiac and skeletal muscle, and the inner ear. This article reviews our current understanding of MiRPs--their functional roles, the mechanisms underlying their association with Kv alpha subunits, their patterns of native expression and emerging evidence of the potential roles of MiRPs in the brain. The ubiquity of MiRP expression and their promiscuous association with Kv alpha subunits suggest a prominent role for MiRPs in channel dependent systems. PMID- 15527816 TI - Activation of Group II and Group III metabotropic glutamate receptors by endogenous ligand(s) and the modulation of synaptic transmission in the superficial superior colliculus. AB - Previous work from this laboratory indicates that Group II/III metabotropic glutamate (mGlu) receptors modulate responses of SC neurones to visual stimuli in vivo. It is thought that tonic levels of glutamate may be sufficient to activate some mGlu receptors. We wished to investigate if these receptors are activated under ambient conditions in SC. Field excitatory postsynaptic potentials (fEPSPs) evoked by optic tract stimulation were recorded from 300 microm slices of the adult pigmented rat superior colliculus at 34 degrees C. The Group II receptor selective agonist LY354740 (100-300 nM) had no significant effect on the peak amplitude of the fEPSP, although it did enhance the late phase of the fEPSP. In order to test for activation of Group II receptors by endogenous ligand, the selective antagonists LY341495 (50 nM) or EGLU (200 microM) were applied: these either enhanced or reduced the fEPSP amplitude. In similar experiments carried out at 22 degrees C, no effect was seen. The fEPSP enhancements, but not the fEPSP reductions, could be occluded by GABA antagonists. Application of higher concentrations of LY341495 (300, 600 nM-known to also affect Group III receptors, particularly mGlu8), or co-application of 50 nM LY341495 and the Group III selective antagonist CPPG (100 microM) produced enhancements of responses, or counteracted response reductions over those seen with 50 nM LY341495 alone. The predominant Group II receptor in SC is mGlu3. It is known that this can be located presynaptically on GABAergic and glutamatergic terminals, postsynaptically, and on glia. Our results indicate that such receptors are tonically activated by endogenous transmitter, have distinct effects, and influence retino-collicular transmission. Furthermore, there is a segregation of effects where receptors exert some of their effects via modulation of GABAergic circuitry. PMID- 15527818 TI - Chronic treatment with imipramine reverses immobility behaviour, hippocampal corticosteroid receptors and cortical 5-HT(1A) receptor mRNA in prenatally stressed rats. AB - Prenatal stress in the rat induces enhanced reactivity of the hypothalamus pituitary-adrenal (HPA) axis, disturbances in a variety of circadian rhythms and increased anxiety-like behaviour. Such abnormalities parallel those found in human depressed patients. Prenatally stressed (PS) rats could represent, therefore, an interesting animal model for the evaluation of the efficacy of pharmacotherapeutic intervention in psychiatric disorders that has often been addressed using control animals. In the present study, PS and non-stressed rats were chronically treated with the tricyclic antidepressant imipramine (10 mg/kg i.p. for 21 days) and assessed in the forced swim test. Glucocorticoid receptor binding sites in the hippocampus were measured and 5-HT(1A) receptor mRNA levels in the frontal cortex were also assessed. PS rats were characterised by increased immobility in the forced swim test, reduced hippocampal corticosteroid receptor binding and increased levels of cortical 5-HT(1A) mRNA. All these parameters were significantly reversed by chronic imipramine treatment. Conversely, no significant effects were observed for non-stressed rats. All these effects are consistent with the expected pharmacotherapy of depression-like abnormalities in PS rats. These results further indicate that PS rats are a relevant animal model of depression. PMID- 15527817 TI - Potentiation of 5-HT3 receptor function by the activation of coexistent 5-HT2 receptors in trigeminal ganglion neurons of rats. AB - 5-HT receptor subtypes are widely expressed in primary sensory neurons, yet so far little is known about the interaction among them. This study aimed to investigate whether the activation of 5-HT2 and 5-HT1 receptors could modulate 5 HT3 receptor mediated current in rat trigeminal ganglion (TG) neurons using whole cell patch clamp technique. The majority of TG neurons examined responded to 5-HT (10(-7)-10(-3) M) with a fast activating and rapid desensitizing inward current (77.2%, 71/92). This 5-HT activated current (I(5-HT)) was blocked by ICS 205-930 and mimicked by 2-methyl-5-HT, indicating that it was mediated by 5-HT3 receptor. With alpha-methyl-5-HT applied prior to 5-HT application, I(5-HT) was potentiated in a concentration-dependent manner, with the maximal modulatory effect at 10(-9) M of alpha-methyl-5-HT. The concentration-response curve for I(5-HT) pretreated with alpha-methyl-5-HT shifts upwards compared with that for I(5-HT) without alpha-methyl-5-HT pretreatment, the maximal I(5-HT) value having increased by (60.3 +/- 5.7)% of its control while the EC50 values of the two curves being very close, i.e. (2.0 +/- 0.3) x 10(-5) M vs (1.7 +/- 0.2) x 10(-5) M, respectively. The alpha-methyl-5-HT potentiation of I(5-HT) was removed by intracellular dialysis of either GDP-beta-S, a non-hydrolyzable GDP analog, or GF109203X, a selective PKC inhibitor, almost completely. Preapplication of R-(+)-UH-301, a selective agonist of 5-HT(1A) receptor, had no modulatory effect on I(5-HT). These results suggest that in the membrane of TG neurons, the activation of 5-HT2 receptors can exert an enhancing effect on the function of coexistent 5-HT3 receptors while that of 5-HT(1A) receptors cannot. PMID- 15527819 TI - Functional responses and subunit composition of presynaptic nicotinic receptor subtypes explored using the novel agonist 5-iodo-A-85380. AB - The novel compound 5-iodo-A-85380 binds with higher affinity to alpha4beta2* nicotinic acetylcholine receptors (nAChR), compared with other nAChR subtypes (Mukhin et al., 2000). In the present study, we have confirmed that in competition binding assays for three major nAChR subtypes, 5-iodo-A-85380 is 850 and 27,000-fold more potent at rat brain alpha4beta2* binding sites than at alpha3beta4 and alpha7 subtypes, respectively. In functional assays, 5-iodo-A 85380 potently activated (EC50 12-35 nM) both alpha-CTx-MII-sensitive and insensitive components of [3H]dopamine release from rat striatal synaptosomes, corresponding to alpha6beta2* and alpha4beta2* nAChR, respectively. 5-Iodo-A 85380 was markedly less potent at eliciting [3H]ACh release from rat interpeduncular nucleus synaptosomes, [3H]noradrenaline release from rat hippocampal slices, and Ca2+ increases in a cell line expressing rat alpha3beta4 nAChR (EC50 = 5, 3.2, 1.6 microM, respectively). As predicted by ligand binding studies, 5-iodo-A-85380 is a more discriminating agonist than the parent compound epibatidine. However, it is not specific for alpha4beta2* nAChR as it also potently activates alpha6beta2* nAChR. PMID- 15527820 TI - The influence of G protein subtype on agonist action at D2 dopamine receptors. AB - In previous studies, we have shown that agonists influence the ability of D2 dopamine receptors to couple to G proteins and here we extend this work. The human D2Short dopamine receptor and a natural polymorphism of this D(2Short)(Ser311Cys), have been studied by co-expressing the receptors in insect cells with Gbeta1gamma2 and either Galpha(o), Galpha(i1), Galpha(i2) or Galpha(i3) G protein subunits. These preparations have been used to study the G protein coupling profiles of the two receptors and the influence of agonists. Receptor/G protein coupling was analysed in dopamine/[3H]spiperone competition binding experiments and through stimulation of [35S]GTPgammaS binding. Although the Ser311Cys polymorphism itself had no appreciable effect on the G protein coupling specificity of the D2 receptor, agonist stimulation of [35S]GTPgammaS binding, revealed that both dopamine and (+)-3PPP showed a clear preference for Galpha(o) compared to the Galpha(i) subtypes, but quinpirole did not. These results indicate that agonists are able to stabilise different receptor conformations with different abilities to couple to G proteins. PMID- 15527821 TI - Effects of nociceptin/orphanin FQ and endomorphin-1 on glutamate and GABA release, intracellular [Ca2+] and cell excitability in primary cultures of rat cortical neurons. AB - The effects of nociceptin/orphanin FQ (N/OFQ) and endomorphin-1 (EM-1) on glutamate and GABA release, intracellular calcium, neuronal excitability and glutamate current were investigated in rat primary cortical neuronal cultures. Through their specific receptors N/OFQ and EM-1 (0.02-1 microM) inhibited the electrically evoked outflow of [3H]D-aspartate at most to -50% and that of [3H]GABA to -30%. In addition, at 1 microM, both peptides induced a decrease of the firing rate caused by electrical depolarization. N/OFQ 1-10 microM did not influence either the electrically evoked calcium influx or the glutamate-evoked currents, whereas EM-1 1 microM significantly inhibited them. Thus, in cortical neurons in culture, both N/OFQ and EM-1 inhibited the secretory process and neuronal excitability but EM-1 also affected calcium influx and cell body responsiveness to glutamate. Consequently, EM-1 appeared to dampen this excitatory signal more then N/OFQ did. PMID- 15527822 TI - Glutamate efflux from human cerebrocortical slices during ischemia: vesicular like mode of glutamate release and sensitivity to A(2A) adenosine receptor blockade. AB - Glutamate extracellular accumulation is an early event in brain ischemia triggering excitotoxic neuron damage. We have investigated how to control the glutamate efflux from human cerebrocortical slices superfused in conditions simulating an acute ischemic insult (oxygen and glucose deprivation). The efflux of previously accumulated [3H]D-aspartate or endogenous glutamate increased starting 18 min after exposure to ischemia and returned almost to basal values in 6 min reperfusion with standard medium. Superfusion with Ca2+-free, EGTA (0.5 mM) containing medium or with medium containing tetrodotoxin (TTX; 0.5 microM) inhibited the ischemia (24 min)-evoked [3H]D-aspartate efflux by about 50% and 65%, respectively. The ischemia (24 or 36 min)-evoked efflux of [3H]D-aspartate or endogenous glutamate was reduced at least 40% by the adenosine A(2A) receptor antagonist SCH 58261 (1 microM); the compound was effective when added up to 15 min after exposure to ischemia. No effect of SCH 58261 on the ischemia-evoked [3H]D-aspartate was found in Ca2+-free, EGTA-containing medium. To conclude, a significant component of the ischemia-evoked glutamate efflux in human cerebrocortical slices seems to occur by a vesicular-like mechanism. Endogenously released adenosine is likely to activate A(2A) receptors that enhance vesicular like glutamate release during ischemia; A(2A) receptor antagonists would deserve consideration for their neuroprotective potential. PMID- 15527823 TI - Neuroprotective effects of the N-terminal tripeptide of insulin-like growth factor-1, glycine-proline-glutamate (GPE) following intravenous infusion in hypoxic-ischemic adult rats. AB - The N-terminal tripeptide of insulin-like growth factor-1, GPE is neuroprotective when given intracerebroventricularly 2 h after hypoxic-ischemic (HI) brain injury in rats. We have now examined whether GPE can cross the blood-brain barrier and exert neuroprotective actions following intravenous administration. Following a single bolus intravenous injection, GPE was rapidly metabolized and cleared from the circulation. The short half-life (<2 min) in blood was subsequently associated with modest and inconsistent neuroprotection. In contrast, potent neuroprotection of GPE was consistently observed in all brain regions examined following 4 h intravenous infusion (12 mg/kg). The neuroprotective effects of GPE after infusion showed a broad effective dose range (1.2-120 mg/kg) and an extended window of treatment to 7-11 h after injury. The central penetration of GPE after intravenous infusion was injury-dependent. GPE also improved long-term somatofunction with a comparable neuronal outcome. GPE reduced both caspase-3 dependent and -independent apoptosis in the hippocampus. Treatment with GPE also inhibited microglial proliferation and prevented the injury-induced loss of astrocytes. In conclusion, the neuroprotective actions of GPE infusion were global, robust and displayed a broad effective dose range and treatment window. GPE's activity included the prevention of neuronal apoptosis, promotion of astrocyte survival and inhibition of microglial proliferation. With injury specific central penetration, GPE has considerable promise as a systemic neuroprotective treatment after acute encephalopathies. PMID- 15527824 TI - Alpha-tocotrienol provides the most potent neuroprotection among vitamin E analogs on cultured striatal neurons. AB - Oxidative stress and apoptosis play pivotal roles in the pathogenesis of neurodegenerative diseases. We investigated the effects of vitamin E analogs on oxidative stress and apoptosis using primary neuronal cultures of rat striatum. A tocotrienol-rich fraction of edible oil derived from palm oil (Tocomin 50%), which contains alpha-tocopherol, and alpha-, gamma- and delta-tocotrienols, significantly inhibited hydrogen peroxide (H2O2)-induced neuronal death. Each of the tocotrienols, purified from Tocomin 50% by high-performance liquid chromatography, significantly attenuated H2O2-induced neurotoxicity, whereas alpha-tocopherol did not. alpha-, gamma- and delta-Tocotrienols also provided significant protection against the cytotoxicity of a superoxide donor, paraquat, and nitric oxide donors, S-nitrosocysteine and 3-morpholinosydnonimine. Moreover, tocotrienols blocked oxidative stress-mediated cell death with apoptotic DNA fragmentation caused by an inhibitor of glutathione synthesis, L-buthionine-[S,R] sulfoximine. In addition, alpha-tocotrienol, but not gamma- or delta-tocotrienol, prevented oxidative stress-independent apoptotic cell death, DNA cleavage and nuclear morphological changes induced by a non-specific protein kinase inhibitor, staurosporine. These findings suggest that alpha-tocotrienol can exert anti apoptotic neuroprotective action independently of its antioxidant property. Among the vitamin E analogs examined, alpha-tocotrienol exhibited the most potent neuroprotective actions in rat striatal cultures. PMID- 15527825 TI - Kynurenine administered together with probenecid markedly inhibits pentylenetetrazol-induced seizures. An electrophysiological and behavioural study. AB - The kynurenine pathway converts tryptophan into various compounds, including l kynurenine, which in turn can be converted to the excitatory amino acid receptor antagonist kynurenic acid, which may therefore serve as a protective agent in such neurological disorders as epileptic seizures. Kynurenic acid, however, has a very limited ability to cross the blood-brain barrier, whereas kynurenine passes the barrier easily. In this study, we tested the hypothesis that kynurenine administered systemically together with probenecid, which inhibits kynurenic acid excretion from the cerebrospinal fluid, results in an increased level of kynurenic acid in the brain that is sufficiently high to provide protection against the development of pentylentetrazol-induced epileptic seizures. CA3 stimulation-evoked population spike activity was recorded from the pyramidal layer of area CA1 of the rat hippocampus, and in another series of behavioural experiments, water maze and open-field studies were carried out to test the presumed protective effect of kynurenine + probenecid pre-treatment against pentylenetetrazol-induced seizures. This study has furnished the first electrophysiological proof that systemic kynurenine (300 mg/kg, i.p.) and probenecid (200 mg/kg, i.p.) administration protects against pentylenetetrazol induced (60 mg/kg, i.p.) epileptic seizures. PMID- 15527826 TI - Caffeine-induced epileptiform field potentials in rat hippocampal slices: a pharmacological characterization. AB - Pharmacological modulation of the epileptiform electric activity induced by caffeine, 10 mM (CAF) on rat hippocampal slices was studied upon field potential recordings in CA3 area of the slices. This concentration of CAF, reportedly releasing Ca2+ ions from the endoplasmic reticulum, led single fimbrial stimuli to evoke repetitive population spikes (PSs) and induced periodic spontaneous field bursts. Carbamazepine, 50 microM reduced (by <40%) the number of repetitive PSs and the rate of spontaneous bursting, with no significant effect on the amplitude of evoked and spontaneous bursts. Valproate, 1 mM reduced only the number (by approximately 25%), but not the amplitudes, of repetitive PSs. Clonazepam, 1 microM consistently reduced the number of repetitive PSs (by approximately 45%), their amplitudes (by 30-60%), and the amplitude of spontaneous bursts (by approximately 70%). The adenosine receptor agonists 2 chloroadenosine, 5 microM and R(-) N6-(2-phenylisopropyl)adenosine, 1 microM had only scanty anti-CAF activity. The depletor of intracellular Ca2+ stores, thapsigargin, 2 microM transiently inhibited the number of evoked PSs and spontaneous bursting. The blocker of ryanodine receptor opening, ruthenium red had an anti-CAF effect, modest at 30 microM, but very strong at 40 microM. Nifedipine, 20 microM opposed CAF-induced spontaneous bursting, but not the evoked PSs. Flunarizine, 50 microM presented only a transient tendency to delay spontaneous bursting. In conclusion, this in vitro slice model appears readily able to reveal antiepileptic properties, though it does not support unequivocal mechanistic interpretation. Nevertheless, anti-CAF activity in this model would suggest the likely involvement of the neuronal ryanodine receptor-related traffic of calcium. PMID- 15527827 TI - Role of intracellular calcium in acute thermal pain perception. AB - The role of intracellular calcium in acute thermal nociception was investigated in the mouse hot-plate test. Intracerebroventricular (i.c.v.) administration of TMB-8, a blocker of Ca++ release from intracellular stores, produced hypernociception. By contrast, i.c.v. pretreatment with thapsigargin, a depletor of Ca++ intracellular stores, produced an increase of the mouse pain threshold. Furthermore, non-analgesic doses of thapsigargin prevented the hypernociception produced by TMB-8. In mice undergoing treatment with heparin, an InsP3-receptor antagonist, or ryanodine, a ryanodine receptor (RyR) antagonist, a dose-dependent reduction of the pain threshold was observed. Pretreatment with D-myo inositol, compound which produces InsP3, and 4-chloro-m-cresol, a RyR agonist, induced an antinociceptive effect. The heparin hypernociception was prevented by D-myo inositol, but not by L-myo inositol, used as negative control. In the same experimental conditions, the antinociception induced by D-myo inositol was prevented by a non-hyperalgesic dose of heparin. Similarly, the reduction of pain threshold produced by ryanodine was reversed by non-analgesic doses of 4-chloro-m cresol, whereas the antinocicpetion induced by 4-chloro-m-cresol was prevented by non-hyperalgesic doses of ryanodine. The pharmacological treatments employed did not produce any behavioral impairment of mice as revealed by the rota-rod and hole-board tests. These results indicate that a variation of intracellular calcium contents at a supraspinal level is involved in the modulation of acute thermal nociception. In particular, the stimulation of both InsP3- and Ry receptors appears to play an important role in the induction of antinociception in mice, whereas a blockade of these receptors is involved in an hypernociceptive response to acute thermal pain. PMID- 15527828 TI - Isolation of SV40 from the environment of a colony of cynomolgus monkeys naturally infected with the virus. AB - The presence of SV40 viral particles in the environment of cynomolgus monkeys naturally infected with this virus has been analyzed by testing waste of the cage samples. SV40 was detected in 2/4 cages tested where mixed infection of SV40 and adenoviruses was observed after inoculation of virions concentrated from cage waste in CV-1 cells. The detected SV40 strains were identical in the regions studied to strain W17, isolated at National Institute for Biological Standards and Control, UK (NIBSC) from a (1/19) monkey kidney biopsy and contains an archetypal regulatory region. The recovery of infectious SV40 virions from the cages provides information about the potential mechanism of transmission of this virus. PMID- 15527829 TI - Replication of SARS coronavirus administered into the respiratory tract of African Green, rhesus and cynomolgus monkeys. AB - SARS coronavirus (SARS-CoV) administered intranasally and intratracheally to rhesus, cynomolgus and African Green monkeys (AGM) replicated in the respiratory tract but did not induce illness. The titer of serum neutralizing antibodies correlated with the level of virus replication in the respiratory tract (AGM>cynomolgus>rhesus). Moderate to high titers of SARS-CoV with associated interstitial pneumonitis were detected in the lungs of AGMs on day 2 and were resolving by day 4 post-infection. Following challenge of AGMs 2 months later, virus replication was highly restricted and there was no evidence of enhanced disease. These species will be useful for the evaluation of the immunogenicity of candidate vaccines, but the lack of apparent clinical illness in all three species, variability from animal to animal in level of viral replication, and rapid clearance of virus and pneumonitis in AGMs must be taken into account by investigators considering the use of these species in efficacy and challenge studies. PMID- 15527830 TI - Myocarditis susceptibility in female mice depends upon ovarian cycle phase at infection. AB - Female BALB/c mice were infected with coxsackievirus B3 in the diestrus, proestrus, estrus, or metestrus phases of the ovarian cycle. Cycle stage was determined by vaginal smear. All mice were killed 7 days after infection. Females infected in the diestrus and especially the proestrus phases developed myocarditis. CD4+ T cells expressing interferon-gamma (IFNgamma) infiltrate the myocardium in these two phases, whereas CD4+ T cells expressing IL-4 are more frequent during estrus. Cardiac virus titers were determined 15 h and 7 days after infection. No differences in virus titer were seen at 7 days. These studies show that natural hormone variations can have substantial effects on viral pathogenicity presumably due to hormone effects on the immune system. PMID- 15527831 TI - Preferential targeting of vesicular stomatitis virus to breast cancer cells. AB - Vesicular stomatitis virus (VSV) is a candidate for development for cancer therapy. We created a recombinant replicating VSV (rrVSV) with an altered surface protein that targeted preferentially to breast cancer cells. The rrVSV genome contained a single glycoprotein (gp) gene derived from Sindbis virus. This gene expressed a chimeric Sindbis E2 binding gp and the native Sindbis E1 fusion gp. The chimeric E2 binding gp, called Sindbis-SCA-erbb2, was modified to reduce its native binding function and to contain a single chain antibody (SCA) with specificity for the human epidermal growth factor receptor Her2/neu protein, erbb2. These viruses selectively infected, replicated in and killed cells expressing erbb2. The titer of rrVSV on SKBR3 cells, a human breast cancer cell line which highly expresses erbb2 was 3.1 x 10(7)/ml compared with a titer of 7.3 x 10(5)/ml on 143 cells, a human osteosarcoma cell line which does not express erbb2. The titer of rrVSV on D2F2/E2 cells, a mouse mammary cancer cell line stably transfected to express human erbb2 was 2.46 x 10(6)/ml compared with a titer of 5 x 10(4)/ml on the parent D2F2 cells which do not express erbb2. When titered on erbb2-negative cells, non-replicating pseudotype VSV coated with Sindbis-SCA-erbb2 had <3% the titer of pseudotype VSV coated with wild type Sindbis gp indicating that the chimeric Sindbis gp had severely impaired binding to the natural receptor. Analysis of the protein composition of the rrVSV found low expression of the modified Sindbis gp on the virus. PMID- 15527832 TI - Sensitivity of prostate tumors to wild type and M protein mutant vesicular stomatitis viruses. AB - Because of its potent ability to induce apoptosis, vesicular stomatitis virus (VSV) is an attractive candidate as an oncolytic virus for tumor therapy. Previous studies have suggested that VSV selectively infects tumor cells due to defects in their antiviral responses making them more susceptible to VSV infection than normal cells. We tested this hypothesis in the prostate tumor system by comparing LNCaP and PC-3 prostate tumor cells to benign human prostatic epithelial cells from patient prostatectomy specimens. We compared the cell killing ability of a recombinant virus containing a wild-type (wt) M protein (rwt) and an isogenic M protein mutant virus (rM51R-M) that induces interferon (IFN) in infected cells and should display a greater selectivity for tumor cells. Our results showed that in single-cycle infection experiments, LNCaP cells were sensitive to killing by both wt and mutant viruses, while PC-3 cells were highly resistant to VSV-induced cell killing. LNCaP and benign prostate cells were similarly susceptible to both viruses, indicating that normal prostate cells are not inherently resistant to killing by VSV. In each of the cell lines, the rM51R M virus induced similar levels of apoptosis to rwt virus, showing that the M protein does not play a significant role in apoptosis induction by VSV in these cells. In multiple-cycle infection experiments, LNCaP cells were more sensitive than benign prostatic epithelial cells to virus-induced cell killing by rM51R-M virus, but not rwt virus. Both viruses were equally effective at reducing LNCaP tumor volume in vivo following intratumoral and intravenous inoculation in nude mice, while PC-3 tumors were resistant to VSV treatment. None of the mice treated with rM51R-M virus died as a result of virus infection, while 50-71% of mice treated with rwt virus succumbed to virus infection. Similarly, when inoculated by the more sensitive intranasal route, the rM51R-M virus was less pathogenic than the rwt virus from which it was derived. These results indicate that M protein mutant viruses are superior candidates as oncolytic viruses for therapies of prostate tumors, but future strategies for use of VSV will require testing individual tumors for their susceptibility to virus infection. PMID- 15527833 TI - Inhibition of certain strains of HIV-1 by cell surface polyanions in the form of cholesterol-labeled oligonucleotides. AB - Cholesterol-labeled oligonucleotides were found several years ago to inhibit HIV 1 in tissue culture at nanomolar concentrations. We present evidence that this is mainly due to an electrostatic interaction between polyanionic oligonucleotide concentrated at the cell surface and a positively charged region in the V3 loop of the HIV-1 envelope protein. When added to tissue culture, cholesterol-labeled oligonucleotides became concentrated at the plasma membrane and potently inhibited virus entry and cell fusion mediated by the envelope protein of some X4 strains of HIV-1, but had little effect on fusion mediated by R5 strains of HIV 1, amphotropic MLV envelope protein, or VSV-G protein. Noncholesterol-labeled oligonucleotides did not bind to the cell surface or inhibit fusion. The pattern of susceptibility to cholesterol-labeled oligonucleotides among HIV-1 strains was the same as reported for nonmembrane-associating polyanions such as dextran sulfate, but the cholesterol-labeled oligonucleotides were effective at lower concentrations. Substitution of a basic 33 amino acid V3 loop sequence from the envelope protein of a resistant strain into a susceptible strain made the envelope protein resistant to inhibition. Inhibition by cholesterol-labeled oligonucleotides was abrogated by the polycation DEAE-dextran. Cholesterol labeled oligonucleotides bound to nonraft regions of the plasma membrane and did not inhibit HIV virus binding to cells. Many infectious agents first associate with target cells via relatively nonspecific charge interactions; our data suggest that molecules that combine a membrane-targeting motif with multiple negative charges might be useful to modify these interactions. PMID- 15527834 TI - VP1 protein of infectious bursal disease virus modulates the virulence in vivo. AB - Infectious bursal disease viruses (IBDVs), belonging to the Birnaviridae family, cause severe immunodeficiency in young chickens by destroying the precursors of antibody-producing B cells in the bursa of Fabricius (BF). Different pathotypes of IBDVs, including cell culture-adapted viruses, differ markedly in virulence, which is characterized by mortality and bursal damage. To study the molecular determinants of virulence in IBDV, the genomic segments A and B of GLS bursa derived (GLSBD) and tissue culture-adapted (GLSTC) viruses were cloned and sequenced. Comparison of the deduced amino acid sequences of segments A and B revealed only two amino acid substitutions at positions 87 (Q --> R) and 261 (P - > L) in segment B, and at positions 253 (Q --> H) and 284 (A --> T) in segment A; the latter of which has been shown to be involved in tissue culture adaptation and attenuation of the virus. To study the function of VP1 protein encoded by segment B, reassortant viruses between tissue culture-adapted strains, GLSTC and D78, and GLSBD were recovered using the reverse genetics system. The recombinant virus rGLSBDB containing segment B of GLSBD was able to replicate in Vero and chicken embryo fibroblast (CEF) cells but exhibited delayed replication kinetics. To evaluate the characteristics of these viruses in vivo, 3-week-old chickens were given equal doses of parental viruses or reassortant viruses by ocular inoculation. The pathological lesions and viral antigen distribution in BF were analyzed at 1, 2, or 3 days postinfection. Parental GLSBD and the recovered rGLSBDB viruses propagate most efficiently in the BF and cause severe bursal lesions, whereas the tissue culture-adapted GLSTC virus replicates less efficiently and induces mild bursal lesions at 3 days postinfection. Taken together, our results demonstrate that the VP1 protein of IBDV is involved in the efficiency of viral replication and modulates the virulence in vivo. PMID- 15527835 TI - Herpes simplex virus 2 VP22 phosphorylation induced by cellular and viral kinases does not influence intracellular localization. AB - Phosphorylation of the herpes simplex virus (HSV) VP22 protein is regulated by cellular kinases and the UL13 viral kinase, but the sites at which these enzymes induce phosphorylation of HSV-2 VP22 are not known. Using serine-to-alanine mutants to map phosphorylation sites on HSV-2 VP22 in cells, we made three major observations. First, phosphorylation by a cellular kinase mapped to serines 70, 71, and/or 72 within CKII consensus sites analogous to previously identified phosphorylation sites in HSV-1 VP22. Second, we mapped UL13-mediated phosphorylation of HSV-2 VP22 to serines 28 and 34, describing for the first time UL13-dependent phosphorylation sites on VP22. Third, previously identified VP22 associated cellular kinase sites in HSV-1 VP22 (serines 292 and 294) were not phosphorylated in HSV-2 VP22 (serines 291 and 293). VP22 expressed alone accumulated in the cytoplasm and to a lesser extent in the nucleus. Phosphorylation by endogenous cellular kinase(s) did not alter the localization of VP22. Co-expression of HSV-2 VP22 with active UL13, but not with enzymatically inactive UL13, resulted in nuclear accumulation of VP22 and altered nuclear morphology. Surprisingly, redistribution of VP22 to the nucleus occurred independently of UL13-induced phosphorylation of VP22. The altered nuclear morphology of UL13-expressing cells was not due to apoptosis. These results demonstrate that phosphorylation of HSV-2 VP22 at multiple serine residues is induced by UL13 and cellular kinase(s), and that the nuclear/cytoplasmic distribution of VP22 is independent of its phosphorylation status but is controlled indirectly by UL13 kinase activity. PMID- 15527836 TI - Determinant for the inhibition of ecotropic murine leukemia virus infection by N linked glycosylation of the rat receptor. AB - Ecotropic murine leukemia viruses (MLVs) recognize the third extracellular loop of the receptor, cationic amino acid transporter type 1 (CAT1). The CAT1 protein contains two conserved N-linked glycosylation sites in the third extracellular loops of the mouse, rat, and hamster receptors (mCAT1, rCAT1, and hCAT1, respectively). Glycosylation of the rCAT1 and hCAT1 receptors inhibits ecotropic MLV infection of CAT1-expressing cells, but that of the mCAT1 does not afford the cells this protection. As compared to the mCAT1 protein, the rCAT1 and hCAT1 proteins possess three and six amino acid insertions, respectively, in the third extracellular loop. To determine whether these inserted amino acids are associated with ecotropic MLV infection inhibition by glycosylation, several mutants of mCAT1 and rCAT1 receptors were constructed. Of all the mutants generated in the present study, only rCAT1 mutant 1 exhibited detectable protein expression levels. The rCAT1 mutant 1-expressing human NP2 cells were more susceptible to transduction by ecotropic MLV vectors than the wild-type rCAT1 expressing cells. Tunicamycin, an N-glycosylation inhibitor, increased transduction titer in the wild-type rCAT1-expressing cells, but did not do so in the cells expressing either the mCAT1 or rCAT1 mutation 1. An amino acid substitution in the glycosylation site of the wild-type rCAT1 conferred higher infection susceptibility, but that of the rCAT1 mutant 1 did not. As with the wild-type mCAT1 and rCAT1 proteins, the rCAT1 mutants were detected on the cell surface by immunofluorescence microscopy. Tunicamycin treatment did not affect cellular distribution of the rCAT1 mutant 1, wild-type mCAT1 or rCAT1 proteins. These results indicate that the extra amino acids in the rCAT1 (as compared to the mCAT1) are associated with inhibition of ecotropic MLV infection by the rCAT1 glycosylation. PMID- 15527837 TI - Species-specific but not genotype-specific primary and secondary isotype-specific NSP4 antibody responses in gnotobiotic calves and piglets infected with homologous host bovine (NSP4[A]) or porcine (NSP4[B]) rotavirus. AB - Using recombinant baculoviruses expressing rotavirus NSP4 [A], [B], [C], and [D] genotypes of bovine, porcine, human, simian, or murine origin, we analyzed serum antibody responses to NSP4s in gnotobiotic calves and piglets infected by the oral/alimentary or intraamniotic route with bovine (NSP4[A]) (Wyatt, R.G., Mebus, C.A., Yolken, R.H., Kalica, A.R., James, H.D., Jr., Kapikian, A.Z., Chanock, R.M., 1979. Rotaviral immunity in gnotobiotic calves: heterologous resistance to human virus induced by bovine virus. Science 203(4380), 548-550) or porcine (NSP4[B]) (Hoshino, Y., Saif, L.J., Sereno, M.M., Chanock, R.M., Kapikian, A.Z., 1988. Infection immunity of piglets to either VP3 or VP7 outer capsid protein confers resistance to challenge with a virulent rotavirus bearing the corresponding antigen. J. Virol. 62(3), 744-748) rotaviruses. Following primary infection and challenge with virulent rotaviruses, the animals developed higher or significantly higher antibody titers to homologous host homotypic NSP4s than to heterologous host homotypic or heterologous host heterotypic NSP4s, indicating that antibody responses were species specific rather than genotype specific. Antibody responses to NSP4s corresponded closely with the phylogenetic relationships of NSP4s within a species-specific region of amino acids (aa) 131 141. In contrast, NSP4 genotypes determined by amino acid full-length sequence identity predicted poorly their "serotypes". In piglets, antibodies to NSP4 induced by previous oral infection failed to confer protection against challenge from a porcine rotavirus bearing serotypically different VP4 and VP7 but essentially identical NSP4 to the porcine rotavirus in primary infection. Thus, in an approach to immunization with a live oral rotavirus vaccine, the NSP4 protein does not appear to play an important role in protection against rotavirus disease and infection. PMID- 15527838 TI - A conserved secondary structure in the hypervariable region at the 5' end of Bamboo mosaic virus satellite RNA is functionally interchangeable. AB - Satellite RNA (satRNA) associated with Bamboo mosaic virus (BaMV) is dependent on BaMV for replication and encapsidation. Molecular analyses of total RNA extracted from bamboo species collected worldwide revealed that 26 out of 61 BaMV isolates harbored satBaMV. Among them, two phylogenetically distinguishable groups, A and B, with a genetic diversity of 6.9 +/- 0.7% were identified. Greatest sequence diversity occurred in the 5' untranslated region (UTR) that contained one hypervariable region with variations of up to 20.7%. Concurrent covariations in the 5' hypervariable sequences support the existence of a conserved apical hairpin stem-loop structure, which was earlier mapped by enzymatic probings and functional analyses [Annamalai, P., Hsu, Y.H., Liu, Y.P., Tsai, C.H., Lin, N.S., 2003. Structural and mutational analyses of cis-acting sequences in the 5' untranslated region of satellite RNA of bamboo mosaic potexvirus. Virology 311 (1), 229-239]. Furthermore, chimeric satBaMVs generated by interchanging the hypervariable region between groups A and B demonstrated the replication competence of satBaMV isolates in Nicotiana benthamiana protoplasts co-inoculated with BaMV RNA. The results suggest that an evolutionarily conserved secondary structure exists in the hypervariable region of 5' UTR of satBaMV. PMID- 15527839 TI - Influence of CD4+ T cell counts on viral evolution in HIV-infected individuals undergoing suppressive HAART. AB - We analyzed the viral C2-V4 envelope diversity, glycosylation patterns, and dS/dN ratios of plasma HIV-1 in an attempt to better understand the complex interaction between viral quasispecies and the host-selective pressures pre- and post-HAART. Phylogenetic analysis of the envelope gene of five patients revealed monophyletic clustering in patients with higher CD4+ T cell counts and sequence intermingling in those with lower CD4+ T cells in relation to the stage of HAART. Our analyses also showed clear shifts in N-linked glycosylation patterns in patients with higher CD4+ T cells, suggesting possible distinct immunological pressures pre- and post-HAART. The relative preponderance of synonymous/nonsynonymous changes in the envelope region suggested a positive selection in patients with higher CD4+ T cells, whereas lack of evidence for positive selection was found in the patients with lower CD4+ T cells. An exception to the last analysis occurred in the only patient who reached complete viral suppression, maybe due to drug pressure exerted over the pol gene that may obscure the immune pressure/selection at the envelope in this analysis. All these indications may suggest that even when HAART generates viral suppression, quasispecies evolve in the envelope gene probably resulting from host-selective pressure. PMID- 15527840 TI - Temperature-sensitive mutants in the vaccinia virus 4b virion structural protein assemble malformed, transcriptionally inactive intracellular mature virions. AB - Two noncomplementing vaccinia virus temperature-sensitive mutants, Cts8 and Cts26, were mapped to the A3L gene, which encodes the major virion structural protein, 4b. The two ts mutants display normal patterns of gene expression, DNA replication, telomere resolution, and protein processing during infection. Morphogenesis during mutant infections is normal through formation of immature virions with nucleoids (IVN) but appears to be defective in the transition from IVN to intracellular mature virus (IMV). In mutant infections, aberrant particles that have the appearance of malformed IMV accumulate. The mutant particles are wrapped in Golgi-derived membranes and exported from cells. Purified mutant particles are indistinguishable from wt particles in protein and DNA composition; however, they are defective in a permeabilized-virion-directed transcription reaction despite containing significant (Cts8) or even normal (Cts26) levels of specific transcription enzymes. These results indicate that the 4b protein is required for proper metamorphosis of IMV from IVN and that proper organization of the IMV structure is required to produce a transcriptionally active virion particle. PMID- 15527841 TI - Evidence that the respiratory syncytial virus polymerase complex associates with lipid rafts in virus-infected cells: a proteomic analysis. AB - The interaction between the respiratory syncytial virus (RSV) polymerase complex and lipid rafts was examined in HEp2 cells. Lipid-raft membranes were prepared from virus-infected cells and their protein content was analysed by Western blotting and mass spectrometry. This analysis revealed the presence of the N, P, L, M2-1 and M proteins. However, these proteins appeared to differ from one another in their association with these structures, with the M2-1 protein showing a greater partitioning into raft membranes compared to that of the N, P or M proteins. Determination of the polymerase activity profile of the gradient fractions revealed that 95% of the detectable viral enzyme activity was associated with lipid-raft membranes. Furthermore, analysis of virus-infected cells by confocal microscopy suggested an association between these proteins and the raft-lipid, GM1. Together, these results provide evidence that the RSV polymerase complex is able to associate with lipid rafts in virus-infected cells. PMID- 15527842 TI - Functional incorporation of green fluorescent protein into hepatitis B virus envelope particles. AB - The envelope of hepatitis B virus (HBV), containing the L, M, and S proteins, is essential for virus entry and maturation. For direct visualization of HBV, we determined whether envelope assembly could accommodate the green fluorescent protein (GFP). While the C-terminal addition of GFP to S trans-dominant negatively inhibited empty envelope particle secretion, the N-terminal GFP fusion to S (GFP.S) was co-integrated into the envelope, giving rise to fluorescent particles. Microscopy and topogenesis analyses demonstrated that the proper intracellular distribution and folding of GFP.S, required for particle export were rescued by interprotein interactions with wild-type S. Thereby, a dual location of GFP, inside and outside the envelope, was observed. GFP.S was also efficiently packaged into the viral envelope, and these GFP-tagged virions retained the capacity for attachment to HBV receptor-positive cells in vitro. Together, GFP-tagged virions should be suitable to monitor HBV uptake and egress in live hepatocytes. PMID- 15527843 TI - Differential ability of two simian virus 40 strains to induce malignancies in weanling hamsters. AB - Different strains of simian virus 40 (SV40) exist and are associated with some human malignancies, but it is not known if SV40 strains differ in biological potential in vivo. In two long-term experiments, Syrian golden hamsters 21 days of age were inoculated by the intraperitoneal route with two different strains of SV40 (10(7) plaque-forming units/animal) and were followed for 8 or 12 months. In vivo responses to strain VA45-54, isolated originally from monkey kidney cells, and to strain SVCPC, recovered from human cancers, were compared. Control animals of the same age were inoculated intraperitoneally with cell culture media. Malignancies developed only in animals infected with SV40 and not in controls. The rate of tumor development was more frequent among animals infected with strain SVCPC than with VA45-54, both in experiments held for 8 months (11/22, 50% vs. 4/20, 20%) and for 12 months (7/15, 47% vs. 3/13, 23%). Histologically, the tumors resembled mesotheliomas, osteosarcoma, and poorly differentiated sarcomas. Metastases to lung and lymph nodes occurred with both viral strains. T-antigen expression was detected in most tumor cells by immunohistochemistry. Anti-T antigen antibodies were produced by almost all tumor-bearing animals and by about two-thirds of those that did not develop tumors after virus inoculation. SV40 viral neutralizing antibodies were detected in all tumor-bearing animals and in 92% and 38% of those inoculated with SVCPC and VA45-54, respectively, that failed to develop tumors. Antibody titers were usually higher in animals with tumors than in those without. Control animals did not develop viral antibodies. Infectious virus was recovered from 2 of 15 tumors tested. This study showed that there are biological differences between these two SV40 strains that influence the outcome of infections in normal hosts, including the development of malignancies and neutralizing antibody, and proved the principle that SV40 strains from different clades can vary in biological properties in vivo. PMID- 15527844 TI - In silico identification of a putative new paramyxovirus related to the Henipavirus genus. AB - A database search for genes encoding paramyxoviral proteins revealed sequences that were designated as human but presented strong evidence of being of viral origin. The two cDNA-derived sequences designated AngRem104 and AngRem52 were originally described as human gene products that were upregulated by angiotensin II in primary mesangial kidney cells. However, their high degree of sequence relatedness to known viral proteins suggests that they represent the P/C/V, M, and F genes of a putative new member of family Paramyxoviridae. Comparison of deduced amino acid sequences and nucleotide motifs suggests that this putative virus is a divergent relative of the Hendra and Nipah viruses; hence, we suggest henipa-like virus or HNLV as a provisional name. Compared to Nipah virus, the percentage of identical (similar) amino acids varied from 19% (42%) for the C protein to 51% (75%) for the M protein. The presence and conservation of presumptive viral transcription start and stop signals and an apparent P editing motif also indicate a relationship of this putative virus to the henipaviruses. Given the highly pathogenic nature of the henipaviruses, the origin of these sequences is enigmatic, and attempts to identify and isolate HNLV are warranted. PMID- 15527845 TI - A Sobemovirus coat protein gene complements long-distance movement of a coat protein-null Dianthovirus. AB - Red clover necrotic mosaic virus (RCNMV; genus Dianthovirus) and Turnip rosette virus (TRoV; genus Sobemovirus) are taxonomically and ecologically distinct plant viruses. In addition, the two genera differ in the role of coat protein (CP) in cell-to-cell movement. However, both are small icosahedral viruses requiring CP for systemic movement in the host vasculature. Here, we show that the TRoV CP gene is capable of facilitating the vascular movement of a Dianthovirus. Substitution of the RCNMV CP gene with the TRoV CP gene permits movement of the resulting chimeric virus to non-inoculated leaves. RCNMV lacking a CP gene or containing a non-translatable TRoV CP gene do not move systemically. This report introduces the molecular characterization of TRoV and describes the unprecedented complementation of systemic movement function by intergenic complete substitution of a plant virus CP gene. PMID- 15527846 TI - Generation and analysis of an RNA vaccine that protects against coxsackievirus B3 challenge. AB - Coxsackievirus B3 (CVB3) is an important human pathogen that causes substantial morbidity and mortality but, to date, no vaccine is available. We have generated an RNA-based vaccine against CVB3 and have evaluated it in the murine model of infection. The vaccine was designed to allow production of the viral polyprotein, which should be cleaved to generate most of the viral proteins in their mature form; but infectious virus should not be produced. In vitro translation studies indicated that the mutant polyprotein was efficiently translated and was processed as expected. The mutant RNA was not amplified in transfected cells, and infectious particles were not produced. Furthermore, the candidate RNA vaccine appeared safe in vivo, causing no detectable pathology following injection. Finally, despite failing to induce detectable neutralizing antibodies, the candidate RNA vaccine conferred substantial protection against virus challenge, either with an attenuated recombinant CVB3, or with the highly pathogenic wt virus. PMID- 15527847 TI - Effect of gene location on the evolutionary rate of amino acid substitutions in herpes simplex virus proteins. AB - In an effort to understand the organization of genes in the herpes simplex virus (HSV-1) genome, I tested the idea that the location of a gene may be related to the evolutionary rate of amino acid sequence variation in the encoded protein. A measure of protein sequence divergence was calculated for homologous proteins in the UL region of six alphaherpesviruses including HSV-1, and this parameter was plotted against position in the HSV-1 genome. The results revealed a cluster of highly conserved proteins (UL27-UL33) encoded near the middle of UL. A similar analysis was restricted to HSV-1 and HSV-2 permitting an examination of U(S) proteins and proteins encoded in repeated regions at the segment ends. This analysis showed that U(S) proteins as a group are more highly divergent than those encoded in UL. A high degree of divergence was also observed in proteins coded at the segment ends including RL1 (gamma(1)34.5), RL2 (alpha0), UL1 (glycoprotein L), UL56, U(S)1, and U(S)12. It is suggested that conserved proteins UL27-UL33 are encoded near the middle of UL to take advantage of a low local mutation rate. Highly divergent proteins are suggested to be encoded selectively in U(S) because of a comparatively rapid evolutionary rate with which genes can be introduced and removed from S in response to environmental variation. PMID- 15527848 TI - Inhibition of human immunodeficiency virus type 1 replication by siRNA targeted to the highly conserved primer binding site. AB - The initiation of HIV-1 reverse transcription occurs at an 18-nucleotide sequence in the viral genome designated as the primer binding site (PBS), which is complementary to the 3' terminal nucleotides of tRNA(Lys,3). Since the PBS is highly conserved among all infectious HIV-1, it represents an attractive target for the development of new therapeutics to inhibit viral replication. In this study, we have evaluated three approaches using small interfering RNA (siRNAs) targeted to the PBS for the capacity to inhibit HIV-1 replication. In the first, transfection of a 21-nucleotide siRNA complementary to the PBS into cells inhibited production of HIV-1 following infection. Control siRNAs of the same length complementary to HIV-1 gag mRNA or to gfp mRNA decreased the production of virus or had no effect on virus replication, respectively. Analysis of the PBS of integrated proviruses derived from viruses that ultimately grew in cultures transfected with siRNA all contained wild-type PBS sequence, demonstrating that HIV-1 did not mutate to escape inhibition by siRNA. In the second approach, hairpin siRNA targeted to the wild-type PBS were expressed using an adeno associated virus (AAV) vector. HIV-1 replication was inhibited in cells infected with AAV encoding the siRNA to the wild-type PBS, but not in cells infected with AAV encoding an siRNA of the same length targeted to an irrelevant PBS. Finally, studies from this laboratory have shown that alteration of the PBS to be complementary to tRNAHis results in the production of infectious virus that rapidly reverts to utilize tRNALys,3 following in vitro culture. A proviral genome containing a PBS complementary to tRNAHis that encodes an siRNA molecule complementary to the wild-type PBS under control of a U6 promoter within the nef gene was as infectious as the parent HIV-1 genome containing no insert in nef. The virus with the PBS only complementary to tRNAHis reverted to use tRNALys,3, coincident with rapid virus growth, while the virus encoding siRNA grew slower than the virus without siRNA and maintained the PBS complementary to tRNAHis longer in culture. At later times of infection, viruses with the PBS complementary to tRNAHis and the siRNA exhibited a rapid increase in p24 antigen in the culture. Analysis of the PBS revealed that it was now complementary to tRNALys,3. Analysis of the gene encoding the siRNA revealed that the reversion of the PBS coincided with the deletion of the gene encoding siRNA. The results of these studies show that siRNA targeted to the PBS of HIV-1 can inhibit virus replication, supporting the concept that HIV-1 has evolved a strong preference to select tRNALys,3 for high-level replication and establishing the PBS and primer selection as a potential target for new therapeutics. PMID- 15527849 TI - Factors limiting the immunogenicity of HIV-1 gp120 envelope glycoproteins. AB - Efficient immune responses to HIV-1 gene products are essential elements to the development and design of an effective vaccine. Ideally, both humoral and cellular responses will be optimally elicited. It is therefore important to elucidate any factors that might limit the immunogenicity of HIV-1 proteins that are likely to be included in an effective vaccine. Since the HIV-1 exterior envelope glycoprotein gp120 is a major target for neutralizing antibodies, it is a virtual certainty that this gene product will be a component of any vaccine that seeks to elicit neutralizing antibody responses from the host humoral immune system. We report here the testing of several HIV-1 gp120 variants derived from a primary isolate that appears deficient in eliciting immune responses at both the level of CD4+ help and consequently in the generation of high-affinity IgG antibody responses in small animals. Factors limiting an effective immune response include (a) envelope glycoprotein strain variation decreasing functional T-cell help, (b) alteration of the glycosylation patterns of gp120 by expression in different cell types, and (c) the native structure of gp120 itself, which may limit the elicitation of effective T-cell help during natural infection or during parenteral immunization in adjuvant. Such limiting factors and others should be considered in the design and testing of gp120-based immunogens in small animals and possibly in primates as well. PMID- 15527850 TI - Vaccination with live attenuated simian immunodeficiency virus for 21 days protects against superinfection. AB - The identification of mechanisms that prevent infection with human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) would facilitate the development of an effective AIDS vaccine. In time-course experiments, protection against detectable superinfection with homologous wild type SIV was achieved within 21 days of inoculation with live attenuated SIV, prior to the development of detectable anti-SIV humoral immunity. Partial protection against superinfection was achieved within 10 days of inoculation with live attenuated SIV, prior to the development of detectable anti-SIV humoral and cellular immunity. Furthermore, co-inoculation of live attenuated SIV with wild type SIV resulted in a significant reduction in peak virus loads compared to controls that received wild-type SIV alone. These findings imply that innate immunity or non-immune mechanisms are a significant component of early protection against superinfection conferred by inoculation with live attenuated SIV. PMID- 15527851 TI - Abrogation of Vif function by peptide derived from the N-terminal region of the human immunodeficiency virus type 1 (HIV-1) protease. AB - The human immunodeficiency virus type 1 (HIV-1) auxiliary gene vif is essential for virus propagation in peripheral blood lymphocytes, macrophages, and in some T cell lines. Previously, it was demonstrated that Vif inhibits the autoprocessing of truncated HIV-1 Gag-Pol polyproteins expressed in bacterial cells, and that purified recombinant Vif and Vif-derived peptides inhibit and bind HIV-1 protease (PR). Here we show that Vif interacts with the N-terminal region of HIV-1 PR, and demonstrate that peptide derived from the N-terminal region of PR abrogates Vif function in non-permissive cells. Specifically, we show that (i) Vif protein binds HIV-1 PR, but not covalently linked tethered PR-PR; (ii) the four amino acids residing at the N terminus of HIV-1 PR are essential for Vif/PR interaction; (iii) synthetic peptide derived from the N terminus of HIV-1 PR inhibits Vif/PR binding; and (iv) this peptide inhibits the propagation of HIV-1 in restrictive cells. Based on these data, we suggest that Vif interacts with the dimerization sites of the viral protease, and that peptide residing at the N terminus of PR abrogates Vif function(s). PMID- 15527852 TI - Contribution of the Gag-Pol transframe domain p6* and its coding sequence to morphogenesis and replication of human immunodeficiency virus type 1. AB - The human immunodeficiency virus type-1 (HIV-1) transframe domain p6* is located between the nucleocapsid protein (NC) and the protease (PR) within the Gag-Pol precursor. This flexible, 68-amino-acid HIV-1 p6* domain has been suggested to negatively interfere with HIV PR activity in vitro proposing a contribution of either the C-terminal p6* tetrapeptide, internal cryptic PR cleavage sites, or a zymogen-related mechanism to a regulated PR activation. To assess these hypotheses in the viral context, a series of recombinant HX10-based provirus constructs has been established with clustered amino acid substitutions throughout the entire p6* coding sequence. Comparative analysis of the mutant proviral clones in different cell culture systems revealed that mutations within the well-conserved amino-terminal p6* region modified the Gag/Gag-Pol ratio and thus resulted in the release of viruses with impaired infectivity. Clustered amino acid substitutions destroying (i) the predicted cryptic PR cleavage sites or (ii) homologies to the pepsinogen propeptide did not influence viral replication in cell culture, whereas substitutions of the carboxyl-terminal p6* residues 62 to 68 altering proper release of the mature PR from the Gag-Pol precursor drastically reduced viral infectivity. Thus, the critical contribution of p6* and overlapping cis-acting sequence elements to timely regulated virus maturation and infectivity is closely linked to precise ribosomal frameshifting and proper N-terminal release of the viral PR from the Gag-Pol precursor, clearly disproving the hypothesis that sequence motifs in the central part of p6* modulate PR activation and viral infectivity. PMID- 15527853 TI - SUMO-1 modification regulates the protein stability of the large regulatory protein Rep78 of adeno associated virus type 2 (AAV-2). AB - The large Rep proteins Rep78 and Rep68 of the helper-dependent adeno associated virus type 2 (AAV-2) are essential for both site-specific integration of AAV DNA in the absence of helpervirus and productive AAV replication in the presence of helpervirus. We have identified UBC9, the E2 conjugating enzyme for the small ubiquitin-related polypeptide SUMO-1, as binding partner of the large Rep proteins in yeast two-hybrid analysis and in GST pulldown assays. Modification of the large Rep proteins with SUMO-1 could be demonstrated in immunoblot analysis and in immunoprecipitations, with the lysine residue at amino acid position 84 serving as the major attachment site. The largely sumolation-deficient Rep78 lysine to arginine point mutant showed a strongly reduced half-life as compared to the wild-type protein. This finding implicates a role for sumolation in the regulation of Rep78 protein stability that is assumed to be critical for the establishment and maintenance of AAV latency. PMID- 15527854 TI - HIV-1 populations in blood and breast milk are similar. AB - Mother-to-child transmission (MTCT) of human immunodeficiency virus type 1 (HIV 1) through breast milk is a significant mechanism of infection in many regions of the world. We compared the HIV-1 populations in paired blood and breast milk samples using a heteroduplex tracking assay (HTA) for the V1/V2 regions of env (V1/V2-HTA). V1/V2-HTA patterns were similar in the eight pairs of samples for which adequate template sampling could be demonstrated. No unique variants existed in either compartment, and differences detected in the relative abundance of variants between compartments were small, occurred among low abundance variants, and were not statistically significant. We also documented the impact of template sampling as a limiting feature in comparing two viral populations. The absence of unique variants and the lack of significant differences in the relative abundance of variants between these compartments support the conclusion that viruses in the blood plasma and breast milk are well equilibrated. PMID- 15527855 TI - An attenuated West Nile prototype virus is highly immunogenic and protects against the deadly NY99 strain: a candidate for live WN vaccine development. AB - In a short time, West Nile virus has developed into a nationwide health and veterinary problem. The high virulence of the circulating virus and related lineage 1 WN strains hinders development of an attenuated live vaccine. We describe an attenuated WN isolate, WN1415, which is a molecularly cloned descendant of the WN prototype B956 strain. The parent virus belongs to lineage 2, members of which have not been associated with epidemic or epizootic outbreaks. A set of non-conservative mutations, mostly in non-structural protein genes, distinguishes the WN1415 isolate from the parent B956 prototype strain. Immunization with WN1415 (55-550,000 pfu) established a potent immunity, which protected the majority of mice against lethal challenge with WN NY99. The attenuated nature of the isolate and its excellent growth characteristics combined with the availability of a highly stable infectious clone make the isolate an attractive candidate for live WN vaccine development. PMID- 15527856 TI - Excess apoptosis of mononuclear cells contributes to the depressed cytomegalovirus-specific immunity in HIV-infected patients on HAART. AB - HIV-infected patients on highly active antiretroviral therapy (HAART) have persistently decreased cytomegalovirus (CMV)-specific proliferative responses [lymphocyte proliferation assay (LPA)] in spite of increases in CD4+ T cell counts. Here we demonstrate an association between apoptosis of unstimulated peripheral blood mononuclear cells (uPBMC) and decreased CMV-LPA. HAART recipients had more apoptosis of uPBMC than controls when measured by caspases 3, 8, and 9 activities and by annexin V binding. Patients with undetectable HIV replication maintained significantly higher apoptosis of CD4+ and CD14+ cells compared to controls. CMV-LPA decreased with higher apoptosis of uPBMC in patients only. This association was independent of CD4+ cell counts or HIV replication. Furthermore, rescuing PBMC from apoptosis with crmA, but not with TRAIL- or Fas-pathway blocking agents or with other caspase inhibitors, increased CMV-LPA in HAART recipients. This effect was not observed in uninfected controls, further indicating that the down regulatory effect of apoptosis on cell-mediated immunity (CMI) was specifically associated with the HIV-infected status. PMID- 15527857 TI - SARS coronavirus E protein forms cation-selective ion channels. AB - Severe Acute Respiratory Syndrome (SARS) is caused by a novel coronavirus (SARS CoV). Coronaviruses including SARS-CoV encode an envelope (E) protein, a small, hydrophobic membrane protein. We report that, in planar lipid bilayers, synthetic peptides corresponding to the SARS-CoV E protein forms ion channels that are more permeable to monovalent cations than to monovalent anions. Affinity-purified polyclonal antibodies recognizing the N-terminal 19 residues of SARS-CoV E protein were used to establish the specificity of channel formation by inhibiting the ion currents generated in the presence of the E protein peptides. PMID- 15527858 TI - Characterization of classical swine fever virus entry by using pseudotyped viruses: E1 and E2 are sufficient to mediate viral entry. AB - Classical swine fever virus (CSFV) is the causative agent of classical swine fever. Its envelope comprises glycoproteins E(rns), E1, and E2. In this study, we showed that the unmodified CSFV glycoproteins could incorporate into the HIV core to generate an infectious CSFV pseudotyped virus. The infection was specific to several porcine cell lines, and could be neutralized by anti-E2 monoclonal antibodies (mAbs) completely and by anti-E(rns) mAbs partially, indicating that this pseudotyped virus can mimic the early infection steps of parental CSFV. To investigate the specific role of each envelope protein involved in viral entry, a series of pseudotyped viruses were generated bearing CSFV glycoproteins in various combinations. It was found that specific infectivity was also achieved with non-E(rns) pseudotyped virus carrying E1 and E2 glycoproteins. This indicated that E1 and E2 are sufficient to mediate CSFV entry, and E(rns) is not indispensable in this process. PMID- 15527859 TI - Emergence of attenuated West Nile virus variants in Texas, 2003. AB - In order to understand how West Nile virus (WNV) has evolved since its introduction into North America, we have studied the genetic and phenotypic variation among WNV isolates collected in various areas during consecutive transmission seasons. The present report describes for the first time phenotypic changes occurring in the North American WNV population. Several isolates collected in Texas during 2003 display a small plaque (sp) and temperature sensitive (ts) phenotype, as well as reduced replication in cell culture, in comparison to isolates collected in 2002 and New York in 1999. Studies of mouse neuroinvasiveness/neurovirulence also indicate that several of these isolates were attenuated in neuroinvasiveness, but not for neurovirulence. The complete genome and deduced amino acid sequences of several of these isolates have been determined in order to map the mutations responsible for this phenotypic variation. These data indicate microevolution of WNV and the emergence of isolates exhibiting phenotypic variation. PMID- 15527860 TI - Effects of sequences of prokaryotic origin on titer and transgene expression in retroviral vectors. AB - Transcriptionally targeted MLV-based ProCon vectors allow expression of the transduced gene in a promoter-specific manner by replacement of the viral U3 region with a heterologous promoter. In order to evaluate the effects of sequence elements present in ProCon vectors on transgene expression (enhanced green fluorescence protein, EGFP), a series of deletion constructs mimicking the situation in proviral DNA following promoter conversion, where expression of the EGFP gene is driven by three different constitutive promoters (MLV U3, mCMV, and hCMV) in the context of a 5'LTR, respectively, were generated and tested in transient transfection experiments. We discovered that modifications in the 3'LTR have only marginal effects on the EGFP expression and the sequence between the promoter and the transgene did not influence EGFP expression at all. On the other hand, EGFP expression was reduced by up to 17-fold in cells transfected with constructs containing SV40neo and/or pBR322ori sequences. To study this effect in transduced cells, we generated a series of retroviral vectors in which these elements were deleted in various combinations and found that an increase in EGFP expression and viral titer was also consistently obtained using vectors lacking these elements, although this was much smaller than that observed using the expression constructs. A vector containing the gene for puromycin resistance (pac) in place of the neomycin resistance gene (neo) was also tested, and found to result in improved vector titers and transgene expression. We conclude that, where possible, the inclusion of neo and ori sequences in retroviral vectors should be avoided, and that, if selection of infected cells is necessary, the pac, rather than neo gene should be used. PMID- 15527861 TI - Sex differences in the vulnerability to drug abuse: a review of preclinical studies. AB - Clinical and preclinical findings indicate that males and females differ on several aspects of drug reinforcement. Females are more vulnerable than males during transition periods of drug use that are characteristic of drug addiction and relapse. Females are also more sensitive than males to the reinforcing effects of stimulants. It has been suggested that ovarian hormones contribute to the mechanisms of action underlying these sex differences. This review examines the preclinical literature on sex differences and ovarian hormonal influences on drug self-administration in animals. It summarizes the findings on the effects of these variables during different phases of drug addiction. Possible differences in the mechanisms of action of drugs of abuse due to interactions with sex differences or ovarian hormonal factors are considered. The animal literature on sex differences in drug abuse treatment effectiveness is also discussed. PMID- 15527863 TI - Serotonin1B receptors: from protein to physiological function and behavior. AB - The serotonin (5-HT)1B receptor is expressed in the central nervous system (CNS) of rodents and its homologous 5-HT1D beta receptor is expressed in human. These receptors are distributed in both serotonergic and non-serotonergic neurons, where they act as auto- or heteroreceptors, respectively. Studies from ours and other laboratories have shown that 5-HT1B receptors are densely expressed in the ventral pallidum, globus pallidus, substantia nigra and dorsal subiculum and moderately expressed in the cerebral cortex, the molecular layer of the hippocampus, the entopeduncular nucleus, the superficial gray layer of the superior colliculus, the caudate putamen and the deep nuclei of the cerebellum. At the ultrastructural level, 5-HT1B receptors were found distributed in axons and axon terminals and these receptors are located on the plasma membrane of unmyelinated axon terminals and in the cytoplasm close to the plasmalemma. The terminal localization of the 5-HT1B receptors in CNS suggests that there is a signal responsible for the protein transport toward the nerve terminals. Studies from ours and other groups using lesion, radioligand binding sites, viral transfection and anterograde methods have shown that 5-HT1B receptors are located at the nerve terminals of different pathways. The 5-HT1B receptors act as terminal receptors and are involved in regulation of the release of various neurotransmitters, including 5-HT itself. The regulation of gamma-aminobutyric acid release by 5-HT1B receptors has been found in projections: from caudate putamen to the globus pallidus or substantia nigra, from nucleus accumbens to the ventral tegmentum area, and from purkinje neurons to the deep nuclei of the cerebellum. The control of glutamate release by 5-HT1B receptors has been found in projections from hippocampus to the dorsal subiculum and of N-acetyl-aspartyl glutamate release from retinal ganglion cells to the superficial gray layer of the superior colliculus. The control of 5-HT release by 5-HT1B receptors was shown in projections arising from the raphe nuclei to fore- and midbrain regions. Multiple evidences suggest that 5-HT1B receptors are implicated in several physiological functions, behavior and psychiatric diseases including migraine, locomotor activity, drug abuse reinforcement, migraine, aggressive behavior, depression and anxiety states. PMID- 15527862 TI - Advances in in vivo imaging of serotonergic neurons in neuropsychiatric disorders. AB - Alterations of central serotonergic neurotransmission, particularly changes in the presynaptically located serotonin transporter (SERT) availability, are thought to be one of the major pathomechanisms of neuropsychiatric symptoms. Modern neuroimaging techniques such as single-photon emission computed tomography and positron emission tomography employ radiolabeled tracers, which bind to SERT, and thus, allow detection of cerebral SERT availability in vivo. We review SERT imaging studies in patients with depression, anxiety disorders, eating disorders and patients with neurological diseases. We furthermore elucidate the potential of SERT imaging techniques in estimating the effect of selective serotonin reuptake-inhibitors. PMID- 15527864 TI - Studies of learning and problem solving in two species of Australian marsupials. AB - This article reviews the authors' recent work with two species of Australian marsupials on several learning and conditioning experiments. The quokka (Setonix brachyurus) is a cat-sized herbivorous wallaby that inhabits offshore islands around south-western Australia. The fat-tailed dunnart (Sminthopsis crassicaudata) is a mouse-sized carnivorous marsupial that inhabits much of inland Australia. Both species were successful in learning simple discriminations, learning sets and reversal sets. Quokkas and dunnarts were also tested on two configural discrimination tasks: transverse patterning and negative patterning; only the dunnarts successfully solved these tasks. Dunnarts were also the only species to show one-trial learning in reversal set training. Dunnarts may be particularly useful subjects in future studies of brain-behavior relationships, since these animals have some interesting neuroanatomical features, demonstrate a wide range of fast learning abilities, and as pouch young, can be accessed during early developmental stages. PMID- 15527865 TI - Does neuropeptide Y (NPY) mediate the effects of psychotropic drugs? AB - Although several studies have summarized the effects of neuropeptide Y (NPY) in the central nervous system, its role in psychopharmacotherapy has not been reviewed in detail. For the last few years, there has been an increase in the number of studies on the suggested role of NPY in the benefits of treatment for mental disorders. Our review focuses on the possible involvement of altered NPY system activity in the effects of antianxiety, antidepressant and antipsychotic therapies. Potential sites and receptors, which are implicated in mediating the NPY effects of psychotropic drugs, have been described. We discuss the significance of alterations in the brain NPY system for the development of new methods of treatment for mental disorders. PMID- 15527866 TI - When is the perirhinal cortex necessary for the performance of spatial memory tasks? AB - The perirhinal cortex and hippocampus have close anatomical links and it has, therefore, been proposed that they have important, coordinated roles in memory. This review examines the relative role of these structures in spatial memory tasks that are known to be hippocampal-dependent. The published lesion data gives a mixed picture, as only some studies detect spatial deficits after perirhinal cortex lesions. The possible reasons for these inconsistencies are reviewed, along with electrophysiological data that indicate how perirhinal cortex lesions may alter neuronal activity in the hippocampus. Overall, the disruptive effects of perirhinal lesions on spatial memory performance are, when they occur, typically transient and never as severe as those seen after hippocampal lesions. It is argued that parallel cortical routes provide key, sensory data to the hippocampus such that in the absence of the perirhinal cortex alternative information is available. The deficits associated with perirhinal damage may then reflect difficulties that arise when task performance requires the use of ambiguous distal cues, for example, those containing overlapping visual features. PMID- 15527867 TI - Molecular genetic aspects of attention-deficit/hyperactivity disorder. AB - Two genome wide scans, one of which was subsequently extended, have led to the identification of different chromosomal regions assumed to harbour genes underlying attention-deficit/hyperactivity disorder (ADHD). Some of these regions were also identified in patients with autism and/or dyslexia. The only region for which both studies detected a LOD score >1 was on chr 5p13 which is in the vicinity of the location of the candidate gene DAT1. The candidate gene approach has revealed the most robust and replicated findings for DRD4, DRD5, and DAT1 polymorphisms. Meanwhile interesting endophenotype studies have also been conducted suggesting a genetic basis for different diagnostic and therapeutic criteria. Animal studies for ADHD have investigated especially hyperactivity and have focused mainly on knockout and QTL designs. In knockout mice models the most promising results were obtained for genes of the dopaminergic pathway. QTL results in rodents suggest multiple loci underlying different forms of natural and induced hyperactivity. The molecular results mentioned above are presented and discussed in detail, thus providing both clinicians and geneticists with an overview of the current research status of this important child and adolescent psychiatric disorder. PMID- 15527868 TI - Thimerosal neurotoxicity is associated with glutathione depletion: protection with glutathione precursors. AB - Thimerosol is an antiseptic containing 49.5% ethyl mercury that has been used for years as a preservative in many infant vaccines and in flu vaccines. Environmental methyl mercury has been shown to be highly neurotoxic, especially to the developing brain. Because mercury has a high affinity for thiol (sulfhydryl (-SH)) groups, the thiol-containing antioxidant, glutathione (GSH), provides the major intracellular defense against mercury-induced neurotoxicity. Cultured neuroblastoma cells were found to have lower levels of GSH and increased sensitivity to thimerosol toxicity compared to glioblastoma cells that have higher basal levels of intracellular GSH. Thimerosal-induced cytotoxicity was associated with depletion of intracellular GSH in both cell lines. Pretreatment with 100 microM glutathione ethyl ester or N-acetylcysteine (NAC), but not methionine, resulted in a significant increase in intracellular GSH in both cell types. Further, pretreatment of the cells with glutathione ethyl ester or NAC prevented cytotoxicity with exposure to 15 microM Thimerosal. Although Thimerosal has been recently removed from most children's vaccines, it is still present in flu vaccines given to pregnant women, the elderly, and to children in developing countries. The potential protective effect of GSH or NAC against mercury toxicity warrants further research as possible adjunct therapy to individuals still receiving Thimerosal-containing vaccinations. PMID- 15527869 TI - Waterborne lead exposure affects brain endocannabinoid content in male but not female fathead minnows (Pimephales promelas). AB - There are several similarities between the behavioral and neurochemical effects of lead (Pb2+) and the cannabinoids. Both Pb2+ exposure and cannabinoid treatment decrease exploratory behavior. Pb2+-induced hyperactivity has been observed in rats and fish. By comparison, cannabinoids increase locomotor activity at higher doses in rats. Moreover, Pb2+ exposure produces learning and memory impairments as do the cannabinoids. Many of the behavioral effects of Pb2+ are thought to be due, in part, to the ability of Pb2+ to either inhibit or mimic the actions of calcium (Ca2+). At low concentrations, Pb2+ enhances basal release of neurotransmitter from presynaptic terminals by increasing intracellular free Ca2+ concentrations. Pb2+ also decreases evoked neurotransmitter release due to blockade of voltage-gated Ca2+ channels. Interestingly, the endocannabinoids (eCBs) including N-arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG) are synthesized in response to increases in intracellular Ca2+ and activate the CB1 receptor that inhibits voltage-gated Ca2+ channels. We tested the hypothesis that waterborne Pb2+ exposure significantly affects whole-brain eCB content in adult male and female fathead minnows (Pimephales promelas). Waterborne Pb2+ exposure (1.0 ppm) resulted in a time-dependent accumulation of Pb2+ in bone in both males and females. Brain AEA and 2-AG content were significantly greater in females compared to males. Pb2+ exposure increased brain AEA content in males at 7 and 14 days of exposure and increased brain 2-AG content at 14 days. Pb2+ exposure had no effect on either brain AEA or 2-AG content in females at any of the time points examined. As eCBs serve as activity-dependent retrograde inhibitors of neurotransmitter release, the increase in brain eCB content would accentuate Pb2+-induced decreases in evoked neurotransmitter release in male but not female fathead minnows. PMID- 15527870 TI - Additional repression of activity-dependent c-fos and BDNF mRNA expression by lipophilic compounds accompanying a decrease in Ca2+ influx into neurons. AB - Recently, it has been proposed that a variety of environmental disruptors (EDs) disturb the neonatal development of the brain in mammals because of their lipophilic characteristics. Therefore, the synergism of these lipophilic compounds is important when evaluating the risk from EDs. In mouse cerebellar granule cells (CGCs), the activity-dependent expression of the brain-derived neurotrophic factor (BDNF) gene is activated through an influx of calcium ions (Ca2+) into CGCs caused by membrane depolarization, which is involved in the activity-dependent development of not only the cerebellum but also other regions of the brain after birth. In our previous study, we reported that permethrin and some other pyrethroid insecticides, which are suspected of being EDs, repressed the induction of c-fos and BDNF mRNA expression, accompanying a reduction of Ca2+ influx at doses non-toxic to CGCs. In the present study, we investigated whether other lipophilic compounds influenced the Ca2+ signal-induced expression of both genes as permethrin did and, if so, whether these effects were synergistic or additional. Pretreatment with p,p'-DDT, diethylstilbestrol (DES) or bisphenol A dose-dependently repressed the induction of both genes as well as the increase in the uptake of Ca2+ by CGCs. Simultaneous exposure of CGCs with permethrin, p,p' DDT and DES, in addition, revealed an additional repression on the induction of the genes and the Ca2+ uptake. These results suggest that toxic effects of EDs might, at least additionally, occur in the brain even if the concentration of each compound is lower than the effective dose for humans. PMID- 15527871 TI - Lysophosphatidylcholine decreases locomotor activities and dopamine turnover rate in rats. AB - Lysophosphatidylcholine (lyso-PTC), a secondary product of S-adenosylmethionine (SAM)-dependent phosphatidylethanolamine (PTE) methylation, is a potent cytotoxin and might be involved in the pathogenesis of Parkinson's disease (PD). Our previous studies showed that the injection of SAM into the brain caused PD-like changes in rodents. Moreover, 1-methyl-4-phenylpyridinium (MPP+), a Parkinsonism inducing agent, increased lyso-PTC formation via the stimulation of PTE methylation pathway. These results indicate a possible role of lyso-PTC in the PD like changes seen following the injection of SAM or MPP+. In the present study, lyso-PTC was injected into the lateral ventricle of rats and locomotor activities and the biogenic amine levels were measured to evaluate the effects of lyso-PTC on the dopaminergic system. Quinacrine, a phospholipase A2 (PLA2) inhibitor, was employed to determine its protective effect on SAM-induced PD-like changes by the inhibition of lyso-PTC formation. The results showed that 1 h after the injection, 0.4 and 0.8 micromol of lyso-PTC increased striatal dopamine (DA) by 20 and 24%, decreased 3,4-dihydroxyphenylacetic acid (DOPAC) by 37 and 45% and decreased homovanilic acid (HVA) by 24 and 13%, respectively. Consequently, dopamine turnover rate, (DOPAC + HVA)/DA, was significantly reduced by 44 and 48% in the rat striatum. Meanwhile, the administration of 0.4 or 0.8 micromol of lyso PTC decreased movement time by 52 and 63%, total distance by 44 and 48% and the number of movements by 43 and 64%, respectively. Quinacrine attenuated SAM induced hypokinesia without affecting SAM metabolism prior to its action on rat brain. The results obtained indicate that the hypokinesia observed following the administration of lyso-PTC might be related to the decline in DA turnover in the striatum in response to lyso-PTC exposure. The present study suggests that inhibitory effects of lyso-PTC on dopaminergic neurotransmission is one of the contributing factors in SAM and MPP+-induced PD-like changes. PMID- 15527872 TI - Soman-induced convulsions in rats terminated with pharmacological agents after 45 min: neuropathology and cognitive performance. AB - It has been demonstrated that a triple regimen consisting of procyclidine (6 mg/kg), diazepam (10 mg/kg) and pentobarbital (30 mg/kg) can effectively terminate soman-induced (1 x LD50) seizures/convulsions in rats when administered 30-40 min following onset. However, convulsive activity lasting for only 45 min can result in marked neuronal pathology. The purpose of the present study was to examine potential cognitive impairments of such brain lesions. The results showed that the neuronal pathology (assessed with Fluoro-Jade B) varied from none at all to 30% damage in the index areas (hippocampus, amygdala, piriform cortex). Cognitive deficits were seen in a novelty test (11 days post-exposure) and retention of a brightness discrimination task (28 days post-exposure) among the rats with neuropathology. Furthermore, significant correlations between neuropathology scores and behavioral measures were found for the animals that convulsed. Among these rats, the mortality rate was relatively high (60%) compared with rats in a previous study that had undergone implantation of hippocampal electrodes (17%). Neither the soman poisoning in the absence of convulsions nor the triple regimen alone affected behavior. It is concluded that early management of soman-induced convulsions is of major importance in preventing neuropathology and accompanying cognitive impairments. PMID- 15527873 TI - Inhibitory effects of cigarette smoke on glial inducible nitric oxide synthase and lack of protective properties against oxidative neurotoxins in vitro. AB - Epidemiological studies consistently report an inverse correlation between cigarette smoking and associated risk for Parkinson's disease (PD). The degeneration of dopaminergic neurons may involve the toxic metabolic products of glial cell monoamine oxidase (MAO) and inducible nitric oxide synthase (iNOS). This study evaluates the direct protective effects of cigarette smoke (CS) against potential neurotoxic products of MAO, such as 1-methyl-4-phenylpyridinium (MPP+), 6-hydroxydopamine (6-OHDA) and hydrogen peroxide (H2O2) in brain neuroblastoma. Moreover, the effects of CS were also evaluated on endotoxin/cytokine activated glioma iNOS protein expression and MAO enzyme activity. Cigarette smoke condensates (CSCs) were acquired from Marlboro 20 Class A and Kentucky 2R4F reference research (2R4F) cigarettes. The CSCs did not protect against 6-OHDA or H2O2 toxicity in neuroblastoma, and exhibited a very mild protective effect [approximately 10%] against MPP+. Neither CSC demonstrated antioxidant capability, but conversely contained high concentration of NO2-. Paradoxically, in glioma cells, iNOS protein expression and endogenous enzymatic NO2- production were significantly blocked by both CSCs. Both CSCs also inhibited glioma MAO-A and MAO-B [1.4.3.4]. Kinetic analysis indicated that 2R4F-CSC displayed competitive inhibition and the Marlboro-CSC exerted potent competitive and non-competitive inhibition. In conclusion, these data suggest that cigarette smoke does not appear to directly protect against the toxicity of the selected neurotoxins. In contrast, CS exerts pronounced effects on glia, whereby its presence can simultaneously attenuate cytokine induction of iNOS and MAO. PMID- 15527874 TI - Modulation of antioxidant defense systems by the environmental pesticide maneb in dopaminergic cells. AB - A lack of evidence supporting a role of heritability in the development of idiopathic Parkinson's disease (PD) has implicated exposures to environmental contaminants in the disease etiology. Epidemiological and clinical studies, as well as animal models of the PD phenotype, have consistently linked agrichemical exposure with dopaminergic (DAergic) damage, particularly through oxidative stress mechanisms. Maneb (MB) is a dithiocarbamate (DTC) fungicide that has specifically been implicated to have adverse effects on dopamine (DA) systems, but the role MB plays in modulating the oxidative state of DAergic cells has not previously been described. Since glutathione (GSH) is a major cellular antioxidant, it was hypothesized that exposure to MB would disrupt this system. The current study primarily utilized the PC12 cell line, which displays a catecholaminergic phenotype. Low concentrations of MB (50-1000 ng/ml) had little effect on cell viability, as measured by LDH release. These same concentrations, however, led to increases in GSH and its oxidized form, GSSG. Effects on viability and GSH were correlated to a primary mesencephalic culture system. Furthermore, these effects were markedly different from those observed with the classical oxidative stressor and pesticide, paraquat (PQ). To determine how MB would affect cells in which antioxidant systems were compromised, PC12 cells were treated with L-buthionine-(S,R)-sulfoximine (BSO) to deplete cellular GSH, followed by treatment with MB. Results suggest that following an insult to the GSH antioxidant system, MB can act as an additional insult to the system and prevent the normal recovery of those defenses. Altered protein levels of heme oxygenase-1 (HO-1) further indicated an oxidative stress response elicited by MB in PC12 cells. DAergic neurons, as a population, are inherently vulnerable to oxidative stress, and the disruption of antioxidant systems by the fungicide MB may contribute to the neurodegeneration of these cells, especially with concurrent exposures to other environmentally relevant oxidative stressors, such as PQ. PMID- 15527875 TI - Chlorpyrifos alters functional integrity and structure of an in vitro BBB model: co-cultures of bovine endothelial cells and neonatal rat astrocytes. AB - The blood-brain barrier (BBB) is a structural and functional interface between the circulatory system and the brain. Organophosphorous compounds such as chlorpyrifos (CPF) may cross the BBB and disrupt BBB integrity and function. To determine events that may contribute to CPF toxicity, we used an in vitro BBB model in which bovine microvascular endothelial cells (BMEC) and neonatal rat astrocytes were co-cultured. We hypothesized that CPF is metabolized by the BBB leading to an inhibition of esterase activity and a disruption of the BBB. The co culturing of BMECs and astrocytes resulted in tight junction formation as determined by electron microscopy, electrical resistance and western blot analysis of two tight junction-associated proteins (ZO-1 and e-cadherin). We observed time dependent increases in ZO-1 and e-cadherin expression and electrical resistance during BBB formation, which were maximal after 9-13 days of co-culturing. The CPF concentration and production of its metabolites were monitored by HPLC following 24 h exposure to CPF on the luminal side of the BBB. We found that the BBB metabolized CPF, with the metabolite 2,3,6-trichloro-2 pyridinol being the major product. CPF and its metabolites were detected on the abluminal side of the BBB suggesting that CPF crossed this barrier. CPF was also detected intracellularly and on the membrane inserts. At tested concentrations (0.1-10 microM), CPF inhibited both carboxylesterase (CaE) and cholinesterase (ChE) activities in BMECs by 43-100%, while CPF-oxon totally inhibited CaE and ChE activity in concentrations as low as 0.1 microM. CPF also caused a concentration-dependent decrease in electrical resistance, with significant inhibition observed at 1 nM and complete loss at 1 microM. These data show that low concentrations of CPF and its metabolites are present within the BBB. CPF and its metabolites, especially CPF-oxon, contribute to the inhibition of CaE and ChE activity, as well as the alteration of BBB integrity and structure. PMID- 15527876 TI - Effect of soman poisoning on populations of bone marrow and peripheral blood cells in mice. AB - According to recent reports, brain lesions resulting from ischemia, mechanical injury or neurodegenerative diseases can be partially treated using bone marrow derived stromal cell (BMSC) engraftment approaches. Nevertheless, for brain lesions resulting from organophosphate poisoning, nerve agents such as soman (pinacolyl methylphosphono-fluoridate) could affect blood and bone marrow (BM) micro-environments, thus preventing efficient BMSC migration and engraftment. It is therefore necessary to verify the hematologic response to soman exposure. To assess this issue, the survival of BM cells, in particular hematopoietic progenitor and precursor cells (HPC), as well as distribution of the different populations of peripheral blood cells, were investigated in soman-intoxicated mice. Nine-week-old adult male B6D2F1 mice were treated with 110 microg/kg soman and 5.0 mg/kg methyl nitrate atropine. BM and peripheral blood (PB) samples were collected 1, 4, 8 and 22 days after poisoning. Various parameters were determined such as PB cell counting or, for BM samples, myelogram, in vitro colony-forming cells and phenotypic flow cytometry analysis. On post-soman day 1, a significant decrease in numbers of white blood cells and an increase in erythrocyte and platelet counts were noted. On post-soman day 4, the number of HPC decreased significantly, probably due to reduction of the replication rate of these immature cells. However, the number of more immature cells (Sca1+/Lin- phenotype) remained unchanged. On post-soman day 8 and day 22, the number of monocytes and granulocytes in the blood had considerably increased, probably due to a strong inflammatory reaction in response to soman poisoning. In conclusion, PB cell and BM-derived HPC populations are affected by acute soman poisoning, suggesting particular care, mainly for graft kinetic aspects, during future development of autologous BM stem cell therapy strategy to treat nerve agent-induced brain damage. PMID- 15527877 TI - Occupations and Parkinson's disease: a multi-center case-control study in South Korea. AB - OBJECTIVE: We performed a hospital based case-control study in South Korea (1) to clarify the role of occupational exposure, and especially manganese (Mn) exposure in the etiology of Parkinson's disease (PD) and (2) to discover the association between any occupations and PD. METHODS: We selected two groups, PD patient group (N1) and controls (N2). Three hundred sixty-seven consecutive outpatients with PD (177 men, 190 women) and 309 controls were interviewed about life style, past history, family history, education level, and occupational history etc. We employed a range of industrial categories as defined by section (the most broad category) and division (sub-category) of the Korea Standard Industry Code (KSIC) Manual. Along with KSIC, we also used the Korea Standard Classification of Occupations (KSCO) as proxies of occupational exposure. The odds ratios (ORs) and 95% confidence intervals (CIs), adjusted for age, sex, smoking status, and education level are presented. RESULTS: As regarding the exposure to hazardous materials, especially Mn, more subjects in the control group than the PD patient group have worked in the occupations with potential exposure to Mn (P < 0.001). Ever having worked in 'agriculture, hunting, and forestry' section of industry was positively associated with PD (OR 1.88), and 'agriculture production crops (OR 1.96)' division of industry was positively associated with PD. On the other hand, ever having worked in the 'manufacturing (OR 0.56)', 'transportation (OR 0.28)' section of industry, and 'transporting (OR 0.20)' division of industry were negatively associated with PD. 'Drivers (OR 0.13)' division of occupation also was negatively associated with PD. CONCLUSIONS: To our knowledge, this is the first case-control studies to find an inverse relationship between 'transporting' or 'technicians like machinery engineers' as his/her longest job and PD risk. Because of this unexpected finding, our work should be replicated in various populations. PMID- 15527878 TI - Blood manganese concentration is elevated in iron deficiency anemia patients, whereas globus pallidus signal intensity is minimally affected. AB - OBJECTIVES: To determine whether blood manganese (Mn) concentration is elevated in patients with iron deficiency anemia (IDA), and whether this affects signal intensities in the globus pallidus. METHODS: Twenty-seven patients with IDA and 10 control subjects were tested for blood Mn, and brain magnetic resonance images (MRI) were also examined. Seventeen of the 27 patients were followed-up after iron therapy. RESULTS: IDA patients had a mean blood Mn concentration of 2.05 +/- 0.44 microg/dl, which was higher than controls. The mean pallidal index (PI) of anemic patients was not different from that of controls. There was a correlation between log blood Mn and PI (rho = 0.384, P = 0.048; n = 27) in IDA patients. None of the patients showed increased signals in the globus pallidus in T1 weighted MRI. Blood Mn levels decreased and hemoglobin levels increased after iron therapy (P < 0.05). CONCLUSION: Although blood Mn is elevated in IDA patients, there is no increase in globus pallidus MRI signal intensity. These findings stand in contrast to those of our other studies showing patients with chronic liver disease or occupational Mn exposure have elevated signal intensities remarkably. PMID- 15527879 TI - Trigeminal uptake and clearance of inhaled manganese chloride in rats and mice. AB - Inhaled manganese (Mn) can enter the olfactory bulbs via the olfactory epithelium, and can then be further transported trans-synaptically to deeper brain structures. In addition to olfactory neurons, the nasal cavity is innervated by the maxillary division of the trigeminal nerve that projects to the spinal trigeminal nucleus. Direct uptake and transport of inhaled metal particles in the trigeminal system has not been investigated previously. We studied the uptake, deposition, and clearance of soluble Mn in the trigeminal system following nose-only inhalation of environmentally relevant concentrations. Rats and mice were exposed for 10-days (6 h/day, 5 days/week) to air or MnCl2 aerosols containing 2.3 +/- 1.3 mg/m3 Mn with mass median aerodynamic diameter (MMAD) of 3.1 +/- 1.4 microm for rats and 2.0 +/- 0.09 mg/m3 Mn MnCl2 with MMAD of 1.98 +/- 0.12 microm for mice. Mn concentrations in the trigeminal ganglia and spinal trigeminal nucleus were measured 2 h (0-day), 7-, 14-, or 30-days post-exposure using proton induced X-ray emission (PIXE). Manganese-exposed rats and mice showed statistically elevated levels of Mn in trigeminal ganglia 0-, 7- and 14 days after the 10-days exposure period when compared to control animals. The Mn concentration gradually decreased over time with a clearance rate (t1/2) of 7-8 days. Rats and mice were similar in both average accumulated Mn levels in trigeminal ganglia and in rates of clearance. We also found a small but significant elevation of Mn in the spinal trigeminal nucleus of mice 7-days post exposure and in rats 0- and 7-days post-exposure. Our data demonstrate that the trigeminal nerve can serve as a pathway for entry of inhaled Mn to the brain in rodents following nose-only exposure and raise the question of whether entry of toxicants via this pathway may contribute to development of neurodegenerative diseases. PMID- 15527880 TI - Neuropathological studies on cycloate-induced neuronal cell death in the rat brain. AB - The herbicide cycloate (carbamothioic acid, ethyl(cyclohexyl)-S-ethyl ester) given as a single oral dose to rats, caused selective neuronal cell death in two regions in the rat forebrain, the pyramidal neurons of layers II-III throughout the pyriform cortex and in granule cells of the caudal ventro-lateral dentate gyrus. Male Alderley Park rats, 6-8-week-old, were given a single oral dose of either 0 or 2000 mg/kg cycloate and killed for neuropathological investigation 1, 2, 3, 7, 14 or 28 days after dosing, using a regime of perfusion fixation with modified Karnovsky's fixative, followed by routine paraffin embedding. Seven transverse levels of brain were examined from each rat. Cycloate-induced neuronal cell death was seen in the pyriform cortex 1 day after dosing and persisted through to Day 28, the lesion was more marked in the rostral compared to the caudal region of the pyriform cortex. Neuronal cell death was also observed in the ventro-lateral caudal dentate gyrus on Days 1-14, day after dosing. In the early stages, Days 1-3 and to a lesser extent Day 7, the neuronal cell death resembled apoptosis, characterized by condensation of nuclear material, cell shrinkage and strong cytoplasmic eosinophilia. By Days 14 and 28 and to a lesser extent Day 7, the cell death resembled necrosis, i.e. karyorrhectic nuclei with pale irregular cytoplasm. Microglial accumulation was associated with the neuronal cell injury. In control brains, an occasional apoptotic body was seen in both the pyriform cortex and dentate gyrus. Our results demonstrate that cycloate is a novel neurotoxicant, which following a single large oral dose induces a cell specific and highly localized forebrain lesion. The time course data analyzed temporally, suggests that cycloate may cause an up regulation of apoptosis in selected regions of the adult brain. PMID- 15527881 TI - Particulate matter in polluted air may increase biomarkers of inflammation in mouse brain. AB - The etiology of neurodegenerative disorders is at present unknown. However, many of these disorders are associated with an increase in oxidative and inflammatory events. Although a small percentage of these disorders are familial cases linked to specific genetic defects, most are idiopathic. Thus, environmental factors are thought to play an important role in the onset and progression of such disorders. We have demonstrated that exposure (4 h, 5 days per week for 2 weeks) to concentrated airborne particulate matter increases inflammatory indices in brain of ovalbumin-sensitized BALB/c mice. Animals were divided into three exposure groups: filtered air (control), ultrafine particles, or fine and ultrafine particles. The levels of proinflammatory cytokines interleukin-1 alpha (IL 1alpha) and tumor necrosis factor alpha (TNF-alpha) were increased in brain tissue of mice exposed to particulate matter compared to that of control animals. Levels of the immune-related transcription factor NF-kappaB were also found to be substantially elevated in the brain of exposed groups compared with the control group. These data indicate that components of inhaled particulate matter may trigger a proinflammatory response in nervous tissue that could contribute to the pathophysiology of neurodegenerative diseases. PMID- 15527882 TI - Inhibition of progenitor cell proliferation in the dentate gyrus of rats following post-weaning lead exposure. AB - Although lead is a potent developmental neurotoxin, the effects of postnatal lead exposure on progenitor cell proliferation in the hippocampus has not been examined. Postnatal day 25 rats were fed a lead containing diet (1500 ppm lead acetate) for 30-35 days and administered bromodeoxyuridine (BrdU, 50 mg/kg, i.p.) during the last 5 days of lead exposure. Animals were killed 24 h after the last BrdU injection. Proliferation of new cells in the subgranular zone and dentate gyrus was significantly decreased in lead-exposed rats compared to control animals that ate a similar diet devoid of lead. These results suggest that postnatal lead exposure can have significant deleterious effects on progenitor cell proliferation and thus the structure and function of the hippocampus. PMID- 15527883 TI - Light-induced reduction of cytoplasmic free calcium in neurons proposed to be encephalic photoreceptors in chick brain. AB - A population of cerebrospinal fluid-contacting neurons (CSFcn) in the lateral septal organ (LSO) may serve as encephalic photoreceptors (EPRs) functioning to signal the onset of seasonal reproductive development in birds. Previous studies on CSFcn in the LSO have focused on identification of retinal protein components in fixed brain tissue. In order to understand better the mechanisms underlying the light-induced photosexual response in birds, a physiological characterization is required. In this study, changes of intracellular free calcium concentration ([Ca2+]i) were monitored during light stimulation of CSFcn in the LSO in live brain slices from embryonic chicks. Using the fluorescent calcium indicator fluo 4, a reduced [Ca2+]i within CSFcn was recorded in response to photostimulation, which is consistent with what has been demonstrated in rods and cones following illumination. Results support the hypothesis that CSFcn in the LSO function as EPRs in the avian brain. PMID- 15527884 TI - Neural stem cell differentiation in a cell-collagen-bioreactor culture system. AB - Neural stem cells and neural progenitors (NSCs/NPs) are capable of self-renewal and can give rise to both neurons and glia. Such cells have been isolated from the embryonic brain and immobilized in three dimensional collagen gels. The collagen-entrapped NSCs/NPs recapitulate CNS stem cell development and form functional synapses and neuronal circuits. However, the cell-collagen constructs from static conditions contain hypoxic, necrotic cores and the cells are short lived. In the present study, NSCs/NPs isolated from embryonic day 13 rat cortical neuroepithelium are immobilized in type I collagen gels and cultured in NASA designed rotating wall vessel (RWV) bioreactors for up to 9 weeks. Initially, during the first 2 weeks of culture, a lag phase of cellular growth and differentiation is observed in the RWV bioreactors. Accelerated growth and differentiation, with the cells beginning to form large aggregates (approximately 1 mm in diameter) without death cores, begins during the third week. The collagen entrapped NSCs/NPs cultured in RWV show active neuronal generation followed by astrocyte production. After 6 weeks in rotary culture, the cell-collagen constructs contain over 10 fold greater nestin+ and GFAP+ cells and two-fold more TuJ1 gene expression than those found in static cultures. In addition, TuJ1+ neurons in RWV culture give rise to extensive neurite outgrowth and considerably more synapsin I+ pre-synaptic puncta surrounding MAP2+ cell bodies and dendrites. These results strongly suggest that the cell-collagen-bioreactor culture system supports long-term NSC/NP growth and differentiation, and RWV bioreactors can be useful in generating neural tissue like constructs, which may have the potential for cell replacement therapy. PMID- 15527885 TI - Nicotine treatment produces persistent increases in amphetamine-stimulated locomotor activity in periadolescent male but not female or adult male rats. AB - Nicotine is a popular addictive drug used among the adolescent population, and it has long been questioned whether nicotine use in adolescence may lead to the use of other psychostimulant drugs. It is not fully understood, however, how nicotine alters behavior and brain neurochemistry in the adolescent age cohort and how this may affect subsequent illicit drug use. In the current study, periadolescent and adult male and female rats were treated with nicotine for 7 days. One day or 30 days after this treatment, the effects of amphetamine on locomotor activity were studied. Sensitization to nicotine occurred in periadolescent female and adult male and female rats, but not in periadolescent male rats over the course of the 7-day treatment period. On day 8 (1 day after treatment with nicotine ended) and on day 37 (30 days after treatment with nicotine ended), nicotine pretreated periadolescent male rats were sensitized to the locomotor-activating effects of amphetamine. The response to amphetamine of periadolescent female and adult male and female rats was unchanged at either time point after nicotine pretreatment. Thus, adolescent males are more sensitive than adults or females to the stimulant effects of amphetamine after exposure to nicotine, and this effect is long-lasting. These data suggest that nicotine use during adolescence may carry a greater risk than during adulthood and that male adolescent smokers may be particularly vulnerable to the risk of stimulant abuse. PMID- 15527886 TI - Hippocampal neurons and recombinant galectins as tools for systematic carbohydrate structure-function studies in neuronal differentiation. AB - Membrane glycoconjugates play a central role in neuronal interactions and regulation. To define the precise links between membrane polysaccharides and neuronal functions, two main requirements must be fulfilled: (1) the availability of molecular tools able to finely discriminate among carbohydrate structures and (2) the use of an experimental system suitable for systematic and quantitative studies of particular neuronal processes. In this work, we used two chicken proto type galectins, i.e., monomeric CG-14 and dimeric CG-16, with very similar carbohydrate affinities, and rat hippocampal neurons in culture to quantitatively measure the involvement of carbohydrate-protein interaction in axonal growth and directionality, neurite sprouting and axon regenerative capacity after section. CG-16 potently stimulated axonal growth and guidance. Neurite sprouting was enhanced by immobilized CG-16 and, notably, reduced by lectin in solution. Overall, cross-linking CG-16 invariably excelled CG-14 in these functional assays, although none of them were able to improve axon regenerative capacity when compared to mammalian galectin-1. Our results demonstrate the potential of the experimental set-up to perform a systematic study of galectin functionality in neuronal differentiation. In view of the concept of the sugar code, the presented results indicate that biological effects triggered by glycan binding engaging an endogenous lectin can be modulated by carbohydrate affinity and/or by other factors like differential cross-linking capacity. PMID- 15527887 TI - Immature developmental pattern of the monosynaptic reflex in isolated spinal cord of glial mutant taiep rats. AB - There is increasing evidence suggesting that glial cells play a crucial role in the formation and maturation of neural circuits. However, little is known about the effects of glial alterations on the establishment of functional circuitry in vivo during the development. The taiep rat, a long-lived neurological mutant characterized by early astrogliosis and demyelination affecting selectively the CNS, provides an interesting model to study the glia-neuron interaction in situ. In the present study, we evaluated the functional development of segmental neural circuits recording the monosynaptic reflex responses (MSR) in the isolated spinal cord of neonatal taiep rats. To evaluate the developmental changes during the first two postnatal weeks, we measured the latency of MSR, the magnitude of depression to paired pulses and the time course of post-tetanic recovery. During the early postnatal period, the MSR of control rats reduced their latency and decreased their sensitivity to depression, as a function of age. By contrast, the MSR of taiep rats failed to develop further from neonatal stage. Near the end of the second postnatal week, the MSR latencies were still prolonged, and the MSR showed a significantly stronger paired pulse depression, and higher post-tetanic recovery times than the age-matched controls. The lack of MSR maturation in taiep rats suggests an early alteration of functional mechanisms underlying the maturation of the spinal reflexes, probably due to the characteristic glial dysfunction(s) of this mutant. PMID- 15527888 TI - Choline exposure reduces potentiation of N-methyl-D-aspartate toxicity by corticosterone in the developing hippocampus. AB - Exposure to high levels of glucocorticoids (GCs) may adversely affect neuronal viability, particularly in the developing hippocampus, via increased function or sensitivity of N-methyl-D-aspartate (NMDA)-type glutamate receptors. Conversely, choline supplementation in the developing brain may reduce the severity of subsequent insult. The present studies aimed to examine the extent to which short term exposure to high concentrations of corticosterone would produce neuronal injury mediated by NMDA receptor activity. These studies also assessed the ability of choline to prevent this form of injury via interactions with nicotinic acetylcholine receptors (nAChRs) expressing the alpha7 subunit. Organotypic hippocampal slice cultures derived from neonatal rat were pre-treated for 72 h with corticosterone (100 nM) alone or with choline (0.1-10 mM), prior to a brief (1 h) NMDA exposure (5 microM). NMDA exposure produced significant cellular damage, reflected as increased fluorescence of the non-vital marker propidium iodide, in the CA1 region. While exposure to corticosterone alone did not produce damage, pre-treatment of cultures with corticosterone markedly exacerbated NMDA induced toxicity. Pre-treatment with choline (> or =1 mM) alone or in combination with corticosterone markedly reduced subsequent NMDA toxicity, effects blocked by co-exposure to methyllycaconitine (100 nM), an antagonist active at nAChRs expressing the alpha7 subunit. These data suggest that even short-term exposure to high concentrations of GCs may adversely affect neuronal viability and that choline supplementation protects the brain from NMDA receptor-mediated damage, including that associated with hypercortisolemia. PMID- 15527890 TI - Distinct periods of cannabinoid sensitivity during zebra finch vocal development. AB - Zebra finch song is a form of vocal communication learned during at least two distinct stages of late postnatal development. During the first of these stages, termed auditory learning, nestlings memorize the song pattern of an adult male tutor, usually the father. During the second stage, sensory-motor learning, these song patterns are practiced and refined until a good copy is produced by adulthood. Vocal learning has made zebra finches a useful model for studying drug effects during vocal development. Prior work has shown that daily exposure to a modest dosage of the cannabinoid agonist WIN55212-2 (WIN) alters sensory-motor learning by reducing stereotypy scores and numbers of note types learned. Here we report that these two effects are produced independently during subperiods of the sensory-motor learning stage. Additional temporally distinct WIN effects during sensory-motor learning include differential incorporation of tutor-derived and improvised note types. We have also evaluated acute and chronic effects of WIN exposure on ability to encode a tutor's song during auditory learning, finding significant effects on stereotypy and distinct effects on note duration and internote intervals. Taken together, these results demonstrate the presence of distinct subperiods of cannabinoid sensitivity during zebra finch auditory and sensory-motor vocal development. PMID- 15527889 TI - Effects of environmental enrichment on behavior and dopamine transporter function in medial prefrontal cortex in adult rats prenatally treated with cocaine. AB - The present study determined if environmental enrichment modifies the effects of prenatal cocaine on open field activity, social interaction and dopamine transporter (DAT) function in the medial prefrontal cortex (mPFC) in rats. Cocaine (40 mg/kg) or saline was administered (s.c.) to pregnant dams from gestation days 8 to 20 (PCOC and PSAL, respectively). At postnatal day 25 (PND 25), female offspring from PCOC and PSAL groups were assigned to the enriched condition (EC; PCOC/EC and PSAL/EC) or impoverished condition (IC; PCOC/IC and PSAL/IC). On PND 60, 90 and 120, locomotor activity, rearing behavior and social interactions were assessed in the open field. On PND 345, rats were anesthetized, challenged with nicotine (0.4 mg/kg), and DAT function in medial prefrontal cortex (mPFC) was assessed using in vivo voltammetry. EC groups displayed decreased locomotor activity across test days, while activity in IC groups did not habituate across days. Generally, PCOC groups displayed more rearing behavior than PSAL groups. During social interaction assessment, IC groups followed their social partner more frequently than EC groups. Moreover, the PCOC/IC group initiated more play solicitations and was engaged in mutual rearing less frequently than PCOC/EC, PSAL/IC and PSAL/EC groups, indicating that epigenetic environmental factors decreased the divergent social behaviors displayed by the PCOC/IC group. Results from in vivo voltammetry experiments demonstrated differences in baseline DAT function in response to environmental enrichment in the prenatal saline groups; however, no effect of prenatal cocaine was observed under baseline conditions. Nicotine challenge unmasked an effect of prenatal cocaine on DA clearance rate in mPFC in the IC groups, which was attenuated by environmental enrichment. Taken together, PCOC/IC rats displayed divergent social interaction and altered DAT function in mPFC, whereas the PCOC/EC group generally was not different from PSAL groups, suggesting that environmental enrichment attenuates the behavioral and neurochemical effects of prenatal cocaine. PMID- 15527891 TI - Ethanol increases retinoic acid production in cerebellar astrocytes and in cerebellum. AB - Several characteristics of fetal alcohol syndrome (FAS) are similar to the teratogenic effects of retinoic acid (RA) exposure. It has been suggested that FAS may result from ethanol-induced alteration in endogenous RA synthesis, leading to abnormal embryonic concentrations of this morphogen. We examined whether ethanol may interfere with RA synthesis in the postnatal cerebellum, as a region of the developing CNS particularly vulnerable to both ethanol and RA teratogenesis. It was found that astrocytes are the predominant source of postnatal RA synthesis in the cerebellum. They express both retinaldehyde dehydrogenase 1 and 2. In vitro cytosolic preparations of astrocytes, as well as live cell preparations, have an increased capacity to synthesize RA in the presence of ethanol. A mechanism by which ethanol could stimulate RA synthesis is via the ethanol-activated short-chain retinol dehydrogenases, which we show to be present in the postnatal cerebellum. To determine whether ethanol stimulated RA synthesis in vivo, a sensitive and highly specific HPLC/MSn technique was used to measure cerebellar RA after administration of ethanol to postnatal day 4 rat pups. Cerebellar RA levels climbed significantly after such treatment. These results suggest that the cerebellar pathology exerted by ethanol may occur, at least in part, through increased production of RA. PMID- 15527892 TI - Neurotrophin expression in the hippocampus and cerebellum is affected by chronic placental insufficiency in the late gestational ovine fetus. AB - Our aim was to determine the effects of chronic placental insufficiency (CPI) during late gestation on the expression of neurotrophic factors and their receptors in the hippocampus and cerebellum in the near-term fetus. Structural alterations were also assessed in these brain regions. CPI was induced in eight fetal sheep by umbilicoplacental embolization (UPE) from 120 to 140 days of gestation (term approximately 147d) such that fetal arterial O2 saturation (SaO2) was maintained at approximately 50% of pre-UPE values. Five non-UPE fetuses served as controls. UPE resulted in fetal hypoxemia, hypoglycaemia, and growth restriction. In hippocampi from UPE fetuses, there were reductions in the optical density (OD) of the immunoreactivity (IR) of brain-derived neurotrophic factor (BDNF) protein within the mossy fibre collaterals of the polymorphic layer and in stratum lucidum (p<0.05); there was no consistent effect on tyrosine-related kinase (Trk) B receptor or neurotrophin-3 (NT-3) expression. Within the cerebellum, there was an increase in BDNF-IR (p<0.05) in the molecular layer; however, Trk B-IR and NT-3-IR were unaltered. There were no significant alterations to the structural parameters measured in the hippocampus. We conclude that CPI in late gestation affects the expression of BDNF in the fetal hippocampus and cerebellum, but these changes do not have a well-defined relationship to structural outcome. PMID- 15527893 TI - Midkine expression in rat spinal motor neurons following sciatic nerve injury. AB - Midkine (MK), a heparin-binding growth factor, is produced in the developing and damaged nervous system. However, the role of MK in peripheral nerve injury has not been clarified. Here, we investigated MK expression in lumbar spinal motor neurons after rat sciatic nerve injury by immunohistochemical, in situ hybridization, and Western blot analyses. The rat sciatic nerve showed complete degeneration after local freezing. Numerous regenerated myelinated and thin nerve fibers were observed 3 weeks after injury. Intense MK immunoreactivity was detected in the ipsilateral spinal motor neurons of the anterior horn of the lumbar spinal cord after 1 day and in ipsilateral and contralateral spinal motor neurons from 4 days to 1 week after injury. It decreased after 2 weeks and again transiently increased in spinal motor neurons after 3 weeks. MK was found in the motor neurons and axon of the sciatic nerve. However, it was not detected in normal neurons and axon. In situ hybridization showed the expression of MK mRNA in lumbar spinal motor neurons of the anterior horn, but it was not present in Schwann cells or non-neuronal cells. Low-density lipoprotein receptor-related protein (LRP) immunoreactivity, a cell membrane receptor of MK, was observed in anterior horn motor neurons, but receptor-type protein tyrosine phosphatase zeta (PTPzeta) immunoreactivity as a signaling receptor complex of MK was not observed. LRP and PTPzeta immunoreactivities were observed in Schwann cells of the injured and uninjured sciatic nerve. Our findings suggest that MK is synthesized, released, and taken up in anterior horn motor neurons in an autocrine fashion with LRP. MK may have a role in degeneration and regeneration after peripheral nerve injury. PMID- 15527894 TI - Characterisation of the Wnt antagonists and their response to conditionally activated Wnt signalling in the developing mouse forebrain. AB - In the present work, the expression patterns of the Wnt antagonists of the Dickkopf (Dkk) family were characterized in the developing mouse forebrain. In situ hybridisation on sections from E12 embryos showed an expression of dkk2 in the thalamus and dkk3 in the cortical hem and thalamus. At later developmental stages (E15.5, E17.5, and P0), little or no expression of dkk1, dkk2, and dkk4 was found in the forebrain, while dkk3 expression was detected in the ventricular zone (VZ) of the lateral and III ventricles, cortical neurons, migrating cells of the primary and secondary dentate migration, and the neuroblastic layer of the eye. In the adult forebrain, dkk3 expression was detected in the lateral VZ, pyramidal neurons of the hippocampus, and cortical neurons. We also provide evidence indicating that only dkk1 and dkk4, along with two other Wnt antagonists axin2 and wif1, but not dkk2 and dkk3, are involved in a feedback mechanism to restrain Wnt signalling in transgenic mice carrying a conditional augmentation of beta-catenin in the forebrain. PMID- 15527895 TI - Ephrin-A5 overexpression degrades topographic specificity in the mouse gluteus maximus muscle. AB - Motor neurons project onto specific muscles with a distinct positional bias. We have previously shown using electrophysiological techniques that overexpression of ephrin-A5 degrades this topographic map. Here, we show that positional differences in axon terminal areas, an entirely different parameter of neuromuscular topography, are also eliminated with ephrin-A5 overexpression. Therefore, we now have both morphological and electrophysiological approaches to explore the mechanisms of neuromuscular topography. PMID- 15527896 TI - In vitro visualization of respiratory neuron activity in the newborn mouse ventral medulla. AB - To clarify the neuronal organization of the respiratory center of the mouse, we analyzed the spatio-temporal pattern of respiratory neuron activity in the ventral medulla of a newborn mouse preparation, using optical recordings. We also demonstrated optical images of the respiratory activity of two different lines of knock-out mice (Tlx3-/-, Pbx3-/-) that exhibit respiratory failure leading to neonatal death from dysfunction of central respiratory neuron activity. In the wild type mice, the respiratory neuron activity in the para-facial region of the rostral medulla appeared prior to inspiratory activity in the more caudal ventrolateral medulla. This rostral to caudal activity pattern was basically preserved in Tlx3-/- mice though the activity was more dispersed and weaker than in the wild type mice. Such an activity pattern was not clearly detected in Pbx3 /- mouse preparations. The difference in the spatio-temporal pattern between Tlx3 /- and Pbx3-/- suggests different levels of functional disorder of the respiratory center. PMID- 15527897 TI - Differential expression of the methyl-cytosine binding protein 2 gene in embryonic and adult brain of zebrafish. AB - Epigenetic gene repression occurs as the result of the interactions between DNA and a number of proteins, including methyl-cytosine binding protein 2 (MeCP2). We have isolated a 1680 bps MeCP2 cDNA from zebrafish that shows deduced amino acid identity with Xenopus and mammalian MeCP2alpha protein sequences. The zebrafish MeCP2 gene was mapped to linkage group 8 using the LN54 radiation hybrid cell panel. The genomic organizations of the zebrafish MeCP2 and mammalian MeCP2alpha are highly similar. Relatively high levels of expression of MeCP2 mRNA were found in embryos at 1 to 4 h postfertilization (hpf), after 24 hpf, and in adult brain and eyes. Whole mount in situ hybridization was performed on embryos and revealed ubiquitous MeCP2 expression during early zebrafish development. At 24 and 48 hpf, the highest levels of expression are found in the epiphysis, midbrain, ventricular zone, and the otic vesicles. In adult zebrafish, MeCP2-expressing cells were found throughout the brain. Regions that are enriched in MeCP2 mRNA include the pallium layer of the telencephalon, the preoptic area, the periventricular grey zone, lobus caudalis, and the vagus lobes. In the cerebellum, high expression is found in the Purkinje and Golgi type 2 cells. PMID- 15527898 TI - Updates on the cytogenetics and molecular genetics of bone and soft tissue tumors: liposarcoma. PMID- 15527899 TI - Evaluation of genomic changes in a large series of malignant ovarian germ cell tumors--relation to clinicopathologic variables. AB - Malignant ovarian germ cell tumors (mOGCT) affect women in their reproductive years, making fertility-saving treatment important. A reliable prediction of the clinical behavior is essential for an optimal therapeutic approach. The genetic changes and molecular mechanisms underlying these rare tumors remain poorly understood. To address these issues, we performed DNA ploidy analysis by high resolution image cytometry in a series of 47 mOGCT and correlated the findings with the DNA copy number changes detected by comparative genomic hybridization (CGH) and clinical outcome. Of 47 tumors, 15 were diploid, 14 were tetraploid, 2 were polyploid, and 13 were aneuploid. All the immature teratomas were diploid, in contrast to the dysgerminomas and endodermal sinus tumors. The International Federation of Gynecology and Obstetrics (FIGO) staging, residual tumors after surgery, and DNA ploidy distribution were significant, independent prognostic factors in survival analysis. The study revealed that the number of DNA copy number aberrations was increased in tetraploid and aneuploid tumors as compared to diploid tumors. Furthermore, a high percentage of aneuploid nuclei in a sample were associated with a complex CGH profile of the tumor in question. The present study confirms that DNA aneuploidy assessment by image analysis may be linked to genetic instability, which is detected as genetic aberrations by CGH. DNA ploidy gives significant prognostic information in addition to the clinical stage in mOGCT with FIGO stage II-IV. PMID- 15527900 TI - High-grade neuroendocrine carcinomas display unique cytogenetic aberrations. AB - Neuroendocrine tumors represent a spectrum of tumor types with different biologic and clinical features. The morphologic types include the low-grade typical and atypical carcinoids and the high-grade small cell and large cell neuroendocrine carcinomas (NECs). Cytogenetic descriptions of high-grade NECs are rare. Complete karyotypic descriptions of 34 high-grade NECs are reviewed: 7 extrapulmonary small cell NECs, 3 metastatic NECs of unknown primary, and 24 small cell lung carcinomas (SCLCs). Chromosomal deletions are more frequent than gains and often involve the entire chromosome arm. Typical aberrations are deletions of chromosome 3p, 5q, 10q, and 17p and gains of 1q, 3q, and 5p occurring as isochromosomes. Non-small cell lung cancers (NSCLCs) have different cytogenetic aberrations, but those with a metastatic phenotype display the identical aberrations as SCLC, a tumor known for its metastatic phenotype at onset. A genetic classification of lung cancer that incorporates the pattern of recurrent chromosome aberrations may be a better predictor of clinical outcome than a morphologic classification. PMID- 15527901 TI - Angiotensin I-converting enzyme gene insertion/deletion polymorphism and endometrial human cancer in normotensive and hypertensive women. AB - Endometrial carcinoma is one of the most common gynecological malignancies. Most cases are diagnosed in older patients with diabetes, hypertension, or obesity. The renin-angiotensin system (RAS) has a central role controlling blood pressure and sodium homeostasis. RAS polymorphisms have been reported as genetic determinants of essential hypertension. The objective of this study was to analyze angiotensin I-converting enzyme gene insertion/deletion polymorphism and endometrial human cancer in normotensive and hypertensive women. The presence of an angiotensin converting enzyme (ACE) polymorphism was analyzed by polymerase chain reaction in DNA isolated from peripheral blood samples of 171 women: 70 cases with endometrial cancer (age, 63.6 +/- 9.5 years) and 101 normal control women (age, 61.3 +/- 6.4 years). We detected DD genotype in 47.5%, ID genotype in 44.3%, and II genotype in 8.2% of cases. The allele frequency was 0.69 for D allele and 0.30 for I allele. In normotensives, we found that the presence of I allele (genotypes ID and II) is significantly associated to an earlier age (56.0 +/- 10.1 versus 65.8 +/- 9.9) of onset of endometrial carcinoma (P=0.029). We observed that normotensive women carriers of an allele I have a higher risk of development of endometrial cancer under the age of 63 years (odds ratio=3.60, 95% confidence interval=1.03-12.56; P=0.037). Our findings suggest that ACE polymorphism may be associated with the development of endometrial carcinoma and with the onset of this tumor in younger women. The definition of a pharmacogenomic profile of human neoplasia may help to identify targets for the development of therapeutic or chemoprevention strategies. PMID- 15527902 TI - Cytogenetic profile in de novo acute myeloid leukemia with FAB subtypes M0, M1, and M2: a study based on 652 cases analyzed with morphology, cytogenetics, and fluorescence in situ hybridization. AB - In about 55% of acute myeloid leukemia (AML) cases, chromosome aberrations are detectable by cytogenetics. Close correlations between cytomorphology and cytogenetics have been reported. To determine a pattern of cytogenetic abnormalities within the French-American-British (FAB) subtypes AML M0, M1, and M2, we analyzed 48 AML M0, 179 AML M1, and 425 AML M2 and compared cytogenetic data to a cohort of 1,062 AML M3/3v, M4, M4eo, M5a/5b, M6, and M7. Cytogenetic abnormalities were significantly more frequent in AML M0 (71%) compared to M1 (49%), M2 (53%), and the total cohort (56%; P < 0.02). While +8 was the most common numeric abnormality in all FAB subtypes, +13, +14, and +11 were associated with AML M0-M2. The only recurring balanced translocation that was associated with one of these FAB subtypes was t(8;21) in M2 (12.5%) and, rarely, M1 (1.7%) (M0, 0% and M3-7, 0.09%; P=0.001). To evaluate the frequency of cytogenetically undetectable abnormalities, we performed fluorescence in situ hybridization (FISH) analyses in 273 AML M0-M2 with normal karyotype using probes for ETO, ABL, MLL, TEL, RB, P53, AML1, and BCR. In two cases we identified numerical aberrations of RB only in interphases nuclei. In seven additional cases, TEL and MLL abnormalities were found. In conclusion, t(8;21), +11, +13, and +14 are strongly associated with AML M0, M1, and M2. The FISH screening analyses identified abnormalities in an additional 3% in normal karyotypes. PMID- 15527903 TI - Genomic organization of the 8p11-p12 amplicon in three breast cancer cell lines. AB - Amplification of chromosomal regions leads to an increase of DNA copy number and expression of oncogenes in human breast cancer (HBC). Amplification of the 8p11 p12 region occurs in 10-15% of primary, uncultured HBCs. In our panel of 11 breast cancer cells, three cell lines, SUM-44, SUM-52, and SUM-225, have overlapping amplicons in the 8p11-p12 region. To characterize genome structure of the amplified regions, we performed fluorescence in situ hybridization using 8p11 p12 BAC clones in the 3 cell lines. The results revealed that the 8p11-p12 amplicon has a highly complex structure and that FGFR1 is not in the common core amplified domain in 3 breast cancer cell lines with the amplicon. These 3 cell lines provide good models for genetic and functional studies of candidate oncogenes of the 8p11-p12 region. PMID- 15527904 TI - M-FISH in gastric lymphoma. AB - The majority of gastric B-cell lymphomas histologically are classified as low grade mucosa-associated lymphoid tissue (MALT lymphoma) and diffuse large B-cell lymphomas (DLBCL). There is evidence that the different histologic types are genetically heterogeneous, evolving through different pathogenetic pathways. Recurrent cytogenetic aberrations have been found in MALT lymphoma, whereas in DLBCL, limited cytogenetic data are available. We report here a DLBCL and a Burkitt-like gastric lymphoma case, cytogenetically studied by G-banding and M FISH technique. In the first case, gains of chromosome 3, 7, 13, and 18 were found. An additional ring chromosome 1 identified as a clonal abnormality suggested clonal evolution. In the second case, trisomy 8, del(6)(q13), as well as t(8;14), t(1;5), and t(1;7), were observed. To our knowledge, cytogenetic data for gastric Burkitt-like lymphoma have not been reported, and M-FISH has not previously been used in the study of gastric lymphomas. PMID- 15527905 TI - Unbalanced translocation der(11)t(11;12)(q23;q13): a new recurrent cytogenetic aberration in myelodysplastic syndrome with a complex karyotype. AB - Cytogenetic abnormalities are observed in approximately one half of cases of myelodysplastic syndrome (MDS). Partial or complete chromosome losses and chromosome gains are frequently found, but there is a relatively high incidence of unbalanced translocations in MDS. We describe here two cases of MDS with an unbalanced translocation, der(11)t(11;12)(q23;q13). Both patients were 69 years of age and diagnosed with refractory anemia with excess of blasts in transformation (RAEB-t) according to the high percentage of blasts in the peripheral blood. Cytoplasmic hypogranulation of neutrophils was evident as a dysplastic change. The blasts were positive for CD4 and CD41a as well as CD13, CD33, CD34 and HLA-DR in both cases. Chromosome analysis showed complex karyotypes including a der(11)t(1;11)(q11;p15)t(11;12)(q23;q13) in case 1 and der(11)t(11;12)(q23;q13) in case 2 plus several marker chromosomes. Spectral karyotyping confirmed the der(11)t(11; 12)(q23;q13) and clarified the origin of marker chromosomes, resulting in del(5q) and del(7q). Fluorescence in situ hybridization (FISH) analyses with a probe for the MLL gene demonstrated that the breakpoints at 11q23 were telomeric to the MLL gene in both cases. FISH also showed that the breakpoint at 11p15 of the case 1 was telomeric to the NUP98 gene. Considering another reported case, our results indicate that the der(11)t(11;12)(q23;q13) is a recurrent cytogenetic abnormality and may be involved in the pathogenesis of advanced-stage MDS. PMID- 15527906 TI - Several chromosomes involved in translocations with chromosome 5 shown with fluorescence in situ hybridization in patients with malignant myeloid disorders. AB - In many patients with myelodysplastic syndromes or acute myeloid leukemia, complex chromosome aberrations can be seen, among which aberrations of chromosome 5 constitute a substantial part. With conventional cytogenetic technique, these aberrations are often identified as deletions or monosomy 5. We analyzed nine patients who, under conventional cytogenetic analysis, showed deletion or monosomy 5. We used fluorescence in situ hybridization with whole-chromosome painting probes to identify the counterpart chromosome and locus-specific identifiers for 5q31 and 5q33 approximately q34. A deletion of 5q was found concomitant with unbalanced translocations. Our results and cases from the literature showed that material from chromosome 5 could be translocated to almost all chromosomes. All patients but one had short survival; this one patient had a preserved 5q31 and 5q33 approximately q34 but a deletion of the q-arm more centromeric than these bands. In eight of the nine patients, further 14 translocations were revealed, not involving chromosome 5. PMID- 15527907 TI - Unusual cytogenetic findings in a synovial sarcoma arising in the paranasal sinuses. AB - A fresh specimen from an intracranial tumor, which was histopatologically classified as a synovial sarcoma, was investigated cytogenetically. The direct method failed to give any information due to a lack of mitotic cells after treatment with 70% acetic acid, whereas 6-day cultures showed a 45,XY,del(6q),-10 karyotype. The histologic evaluation was consistent with a synovial sarcoma, which is well characterized from a cytogenetic perspective and involves a t(X;18)(p11;q11) in more than 95% of cases. Reverse-transcription polymerase chain reaction analysis did not show the presence of the SYT-SSX chimeric gene. PMID- 15527908 TI - Gluthatione-S-transferase P1 polymorphism I105V in familial and sporadic prostate cancer. AB - Several reports suggest that the glutathione-S-transferase (GST) family of enzymes is involved in a variety of cancers, due to their carcinogen detoxification properties. A polymorphism in codon 105 of the pi variant (GSTP1 I105V), which affects the enzymatic activity of the enzyme, has been linked to the incidence of cancers from different organs. However, the published data in prostate cancer (PCa) is controversial. Some studies report an association with the GSTP1 I105V polymorphism and sporadic PCa, whereas other studies report no association. Recently, one study showed a positive correlation between the GSTP1 I105V polymorphism and familial PCa in a Japanese population. In the present study, we assessed the correlation of the GSTP1 I105V polymorphism with familial and sporadic PCa in an American population. We analyzed DNA samples from 438 patients with familial PCa, 499 patients with sporadic PCa, and 510 controls. We found no significant association between the GSTP1 I105V polymorphism and familial or sporadic PCa when compared to the control group [odds ratio (OR) =1.0 (0.74-1.37); P=0.58]. Moreover, no association was found after stratification for age of diagnosis, Gleason grade, or lymph node involvement [OR =0.84 (0.65-1.09), P=0.37]. These data indicate that there is no associated risk for sporadic or familial PCa in American families containing the GSTP1 I105V polymorphism. PMID- 15527909 TI - A malignant lymphoma of the thyroid with trisomy 21 as the sole chromosomal anomaly. PMID- 15527910 TI - Combination of toluidine blue staining and in situ hybridization to evaluate paraffin tissue sections. PMID- 15527911 TI - Met688Ile and Leu390Phe of the MSH2 gene are not functional mutations, but polymorphisms in Japanese individuals. PMID- 15527912 TI - Growth inhibition of foodborne pathogens and food spoilage organisms by select raw honeys. AB - Twenty-seven honey samples from different floral sources and geographical locations were evaluated for their ability to inhibit the growth of seven food spoilage organisms (Alcaligenes faecalis, Aspergillus niger, Bacillus stearothermophilus, Geotrichum candidum, Lactobacillus acidophilus, Penicillium expansum, Pseudomonas fluorescens) and five foodborne pathogens (Bacillus cereus, Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica Ser. Typhimurium, and Staphylococcus aureus) using an overlay inhibition assay. They were also tested for specific activity against S. aureus 9144 and B. stearothermophilus using the equivalent percent phenol test--a well diffusion assay corresponding to a dilute phenol standard curve. Honey inhibited bacterial growth due to high sugar concentration (reduced water activity), hydrogen peroxide generation, and proteinaceous compounds present in the honey. Some antibacterial activity was due to other unidentified components. The ability of honey to inhibit the growth of microorganisms varies widely, and could not be attributed to a specific floral source or demographic region produced in this study. Antibacterially active samples in this study included Montana buckwheat, tarweed, manuka, melaleuca, and saw palmetto. Furthermore, the bacteria were not uniformly affected by honey. Varying sensitivities to the antimicrobial properties were observed with four strains of S. aureus thus emphasizing the variability in the antibacterial effect of honey samples. Mold growth was not inhibited by any of the honeys tested. B. stearothermophilus, a heat-resistant spoilage bacteria, was shown to be highly sensitive to honey in both the overlay and well diffusion assays; other sensitive bacteria included A. faecalis and L. acidophilus. Non-peroxide antibacterial activity was observed in both assays; the highest instance was observed in the specific activity assay against B. stearothermophilus. Further research could indicate whether honey has potential as a preservative in minimally processed foods. PMID- 15527914 TI - Antimicrobial susceptibility of vancomycin-susceptible and -resistant enterococci isolated in Italy from raw meat products, farm animals, and human infections. AB - The susceptibility of vancomycin-resistant (VRE) and vancomycin-susceptible (VSE) enterococci to 10 antimicrobial agents was evaluated. The strains, belonging to different species, were isolated in Italy from raw meat products, farm animals, and human clinical infections in the years 1997-2000. High frequency of resistance to tetracycline and erythromycin was observed in all the groups of strains. On the contrary, chloramphenicol was the only drug that showed a relatively low rate of resistance in all the groups examined. In general, the resistance rates observed for VSE did not differ from those observed for VRE of the same species and origin. Some differences could be noticed among the different enterococcal species, with Enterococcus faecium strains being usually more resistant to beta-lactams, and Enterococcus faecalis strains more resistant to gentamicin. However, the strongest differences were observed when the strains were compared according to their source, the human isolates being usually more resistant than the isolates of animal origin. No significant difference was observed between isolates of swine and poultry origin. Among VRE E. faecium, multiple resistance was much more frequent among the human strains (90%) than among poultry (48.9%) and swine (26.5%) strains. These results show that in Italy VRE isolates from human clinical infections are usually more resistant than isolates from meat products and farm animals, and possess different antimicrobial resistance profiles. PMID- 15527913 TI - Two homologous oligopeptide binding protein genes (oppA) in Lactococcus lactis opp2 [corrected]. AB - In previous studies, it has been shown that inactivation of opp or even oppA abolishes the capacity of Lactococcus lactis to utilize oligopeptides. We now show that the opp operon has been duplicated in L. lactis MG1363. The nucleotide sequence of the oppA and oppC homologues (appA and appC) and most of the oppB homologue (appB) indicate that the corresponding protein sequences are 83%, 92% and 91% identical, respectively. Inactivation of appA, via homologous recombination, as well as complementation studies were carried out to determine the possible function of appA in peptide utilization. As anticipated from studies with an oppA knock-out, peptide utilization was not impaired in an appA disruption mutant. Importantly, AppA expressed from a plasmid could restore the ability of oppA deletion mutants to utilize Leu-enkephalin, albeit with a lower efficiency than OppA. The differences in the ability to utilize this pentapeptide were not due to differences in expression levels but most likely reflect a different catalytic efficiency in oligopeptide utilization when AppA is used as ligand receptor. PMID- 15527915 TI - Antimicrobial resistance profile of five major food-borne pathogens isolated from beef, pork and poultry. AB - This study was performed to evaluate the resistance rate against antimicrobials of food isolates of the five major food-borne pathogens to compare these and to possibly distinguish a pattern. A total of 922 samples of the major meat species (pork, beef and poultry) were analysed for thermophilic Campylobacter, Salmonella, Yersinia enterocolitica, pathogenic Escherichia coli and Listeria monocytogenes. Isolates were subjected to antimicrobial resistance testing by the disc diffusion method. Roughly the same overall rate of resistance was identified for thermophilic Campylobacter, Salmonella and pathogenic E. coli. Resistance to quinolones and tetracycline was determined most frequently. In contrast, food isolates of Y. enterocolitica and L. monocytogenes were rarely tested resistant. The significance of our findings is that resistance rates in enteric bacteria seem to be much higher than in pathogens found in a variety of environments, closely associated to the host environment. PMID- 15527916 TI - The pH-unrelated influence of salt, temperature and manganese on aroma formation by Staphylococcus xylosus and Staphylococcus carnosus in a fermented meat model system. AB - The influence of manganese (0.01-0.1-1.0 microg/g), temperature (15-24 degrees C) and salt (3-4% w/w) on volatile formation in model minces inoculated with Pediococcus pentosaceus and either Staphylococcus xylosus or Staphylococcus carnosus was studied in a full factorial experiment. In order to study the direct, pH-unrelated effect of the parameters, data were analysed by use of multiple linear regression and partial least-squares regression both before and after transformation of the volatile responses into pH-orthogonal (pH-unrelated) responses. By using the pH-orthogonalised data, the overall interpretability of the experiment was increased, and new cause-and-effect relations were suggested. Approximately 50% of the total variance in volatile levels was due to differences caused by S. xylosus and S. carnosus, and another 30% was related to differences in pH development. The remaining 20% covered pH-orthogonal effects of manganese, temperature and salt plus the experimental noise. From this, it was concluded that most of the variation in volatile profiles caused by manganese, temperature and salt was in fact directly or indirectly caused by changes in lactic acid bacterial activity and pH. PMID- 15527917 TI - A collaborative study to evaluate qualitatively powdered baby food validation samples artificially contaminated with Salmonella anatum. AB - Nineteen laboratories participated in a collaborative study to evaluate the performance of validation samples consisting of powdered infant formula and cereal-based weaning food artificially contaminated with low levels of Salmonella anatum. The Standard method BS EN ISO 6579:2002 was to be followed for the trial. Salmonella counts in each baby food were around 10 CFU/25 g (low) or 10 CFU/g (high level). Trial participants received five samples of each sample type and five blank samples. All samples contained Pseudomonas aeruginosa and Lactobacillus spp. at concentrations between 10(4) and 10(5) CFU/g as background flora. Precision data for the powdered infant formula were similar for the solid selective plating-out medium xylose-lysine-deoxycholate (XLD) and a 2nd choice of agar suitable to isolate Salmonella spp. The sensitivity was 91.3%, accordance 82.5% and concordance 83.9% for the low and 100% for the high level in all cases. For the cereal-based weaning food, the precision data for the high level were similar for XLD and the 2nd choice medium (98.8%, 97.5% and 97.5%). A slight difference was observed for the low level where XLD obtained values of 82.5%, 73.8%, 70.6% for sensitivity, accordance and concordance and the second choice medium values of 81.5%, 72.5% and 69%. The specificity, accordance and concordance of blanks for the infant formula and cereal-based weaning food were 100%. PMID- 15527918 TI - Microbial profiles of commercial, vacuum-packaged, fresh pork of normal or short storage life. AB - The microbial ecology of fresh vacuum-packed pork cuts during storage at -1.5 degrees C for up to 45 days was examined to characterize rates of microbial growth and pH changes in commercially prepared products of normal storage quality. Pork loins in commercial distribution with odour defects were also studied to determine a possible cause of the defects and avoid future problems. In addition, microbial profiles of pork cuts from two plants were compared, after storage for 25 days at -1.5 degrees C, to identify possible reasons for differences in the storage life of product from the plants. The effects of a change in sanitation procedures on the microbial populations of products stored for 25 days were also studied. With normal product, microbial growth in different packages progressed at different rates, reflecting differences in initial levels of bacterial contamination. All samples in the study reached 8 weeks without apparent organoleptic change and samples carried 5.8+/-1.2 log bacteria cm(-2) (mean+/-S.D.). The flora of loins with the odour defect were predominately lactic acid bacteria (LAB) and carnobacteria, but they contained large fractions of Enterobacteriaceae <35 days after packaging. Aeromonas spp. and Shewanella spp. were likely responsible for the sulfide-putrid smell of these spoiled products, but species of Enterobacteriaceae and lactic acid bacteria could have contributed to spoilage. Comparison of microbial groups present in 16 other cuts, half from each of two commercial plants, which were stored for 25 days at -1.5 degrees C, showed that larger fractions of Enterobacteriaceae were present in samples from the plant having difficulty achieving the desired storage life. Additional bacterial samples from 12 cuts supplied by the latter plant obtained after adoption of an acid sanitizer step in the plant cleaning regimen, and also stored for 25 days at -1.5 degrees C, yielded few Enterobacteriaceae, Aeromonas or Shewanella. Use of an acid sanitizer in plant cleaning may be a means of controlling alkali-tolerant bacteria such as Aeromonas or Shewanella which can contaminate pork cuts and spoil vacuum-packaged product. The fraction of Enterobacteriaceae in bacteria populations on fresh pork stored for 25 days at 1.5 degrees C may be a useful indicator of the effectiveness of plant sanitation. PMID- 15527919 TI - Antimicrobial effect of water extract of sumac (Rhus coriaria L.) on the growth of some food borne bacteria including pathogens. AB - The antimicrobial effect of water extracts of sumac (Rhus coriaria L.) at concentrations of 0.1%, 0.5%, 1.0%, 2.5% and 5.0% (w/v), non-neutralized and after neutralization to pH 7.2+/-0.1, was studied on the growth of 12 bacterial strains (six Gram positive strains and six Gram negative strains), mostly food borne including pathogens. It was found to be effective against all the test organisms with Gram positive strains being more sensitive than Gram negative strains. Significant differences (P<0.01) were found among the bacteria and between the non-neutralized and neutralized extracts with non-neutralized being more effective against all the bacteria. The minimal inhibitory concentration (MIC) of the extract for each bacterial strain was studied by a gradient plate method. Among the Gram positive organisms, Bacillus species (Bacillus cereus, Bacillus megaterium, Bacillus subtilis, and Bacillus thuringiensis) were found to be the most sensitive showing MICs of 0.25-0.32% (after 24 h incubation) followed by Staphylococcus aureus (0.49%), while Listeria monocytogenes was found to be the least sensitive demonstrating a MIC of 0.67%. Of the Gram negative organisms, Salmonella enteritidis was found to be the most resistant with a MIC of 0.67% followed by Escherichia coli Type I, E. coli O157:H7, Proteus vulgaris and Hafnia alvei having MICs of 0.63%, 0.60%, 0.55% and 0.45%, respectively; whereas Citrobacter freundii was found to be the least resistant surviving up to 0.42%. Some loss of antimicrobial activity was, however, observed after incubation for 3 days. Bacteriostatic/bactericidal effects of sumac, as studied by enumerating survival by the viable count technique after 1 h direct contact of each microorganism with various concentrations of sumac extract, revealed a 4-5 log cycle reduction in Bacillus spp. and 2-3 log cycle reduction in other bacteria tested with 1.0% sumac extract. PMID- 15527920 TI - Behavior of Clostridium perfringens at low temperatures. AB - Refrigerated storage is an important step in the preparation of foods and inadequate storage is one of the main causes of food poisoning outbreaks of Clostridium perfringens. Therefore, growth and germination characteristics of C. perfringens in a temperature range of 3-42 degrees C were determined in fluid thioglycollate broth (FTG) and Dutch pea soup. To study the effect of adaptation, cells were either inoculated from a 37 degrees C pre-culture or from a temperature-adapted pre-culture. Membrane fatty acid patterns were determined at all temperatures to examine the effect of temperature on membrane composition. Spores were either inoculated with and without heat treatment. Adaptation of cells did not influence growth rate nor lag phase. Growth in pea soup, however, was slower and lag phases tended to be more extended compared to FTG. No growth was observed at temperatures < or =10 degrees C and death rates in pea soup were higher than those in FTG at these low temperatures. Cells preserved the membrane fluidity by reducing the arachidic acid content and increasing the lauric acid content when the temperature dropped. This resulted in a net reduction in chain length. Microscopic analysis of cells grown at 15 degrees C revealed a morphological change: cells were elongated compared to those grown at 37 degrees C. These data demonstrate the ability of C. perfringens to adapt to lower temperatures. However, this did not influence growth characteristics compared to non-adapted cells. Spores of C. perfringens did germinate at all temperatures with and without heat-activation. Combining this fact with the extended survival at low temperatures emphasizes the need for adequate heating of refrigerated foods before consumption to eliminate health risks due to C. perfringens. PMID- 15527921 TI - Isolation of shiga toxigenic Escherichia coli from raw beef in Palestine. AB - Shiga toxigenic Escherichia coli (STEC) isolated from raw beef samples in northern Palestine during a 1-year period were characterized for virulence genes by a polymerase chain reaction (PCR) assay and screened for their antibiotic resistance. STEC was identified in 44 (14.7%) of 300 raw beef samples. Twelve (27.3%) of the STEC isolates were serotype O157. Nine of those were isolated during summer. The majority of STEC isolates (70.5%) harbored both stx1 and stx2 genes, while the others harbored either stx1 or stx2. High levels of resistance against different antimicrobial agents were detected. Resistance to at least three drugs was found in 55% of the isolates. PMID- 15527922 TI - Growth inhibition of heat-injured Enterococcus faecium by oligophosphates in a cured meat model. AB - Cells of two heat-resistant strains of Enterococcus faecium were heated and incubated in meat suspensions containing curing ingredients. The concentrations of the curing ingredients were those frequently used for pasteurized ham-type products, except that the concentrations of the oligophosphates (triphosphate and diphosphate) varied. Heating tests at 69 degrees C were performed with inoculated meat suspensions in heat-sealed plastic pouches. Numbers of bacteria were counted immediately after heating and in parallel series of heated pouches incubated at 37 degrees C. Plating was performed in Tryptone Dextrose Yeast Meat Peptonised Milk Agar (TDYMP); in TDYMP Agar to which the curing ingredients were added; and in TDYMP Agar to which the curing ingredients except oligophosphates were added. The inclusion of oligophosphates in the heating medium increased the heat-injury sustained by the E. faecium cells, and in combination with rather severe heat treatment even completely blocked the growth of surviving organisms in the meat suspension incubated at 37 degrees C. The presence of oligophosphates in the culture medium TDYMP Agar severely reduced the counts of freshly heated cells; however, this effect disappeared after repair and growth of the surviving organisms in the meat suspension. PMID- 15527923 TI - Development of culture media containing spent yeast cells of Debaryomyces hansenii and corn steep liquor for lactic acid production with Lactobacillus rhamnosus. AB - To develop a cost-effective fermentation medium, biomass of Debaryomyces hansenii coming from xylitol production and corn steep liquor was evaluated for lactic acid production by Lactobacillus rhamnosus in glucose-containing media. Glucose consumption and lactic acid generation were followed using a variety of media made with different nutrient supplementation. Under selected conditions, media containing glucose, biomass of D. hansenii and corn steep liquor as unique components led to product yields similar to those obtained in a fully supplemented medium. PMID- 15527924 TI - Relationships of ratings of appetite to food intake in healthy older men and women. AB - The aim of this study was to determine how rated appetite relates to the amount eaten in a meal in healthy older people. On two study days, 32 healthy older men (n = 16) and women (n = 16) aged 65-85 years, recruited by advertisement, consumed a standardised breakfast and 4 h later were offered lunch from which they could eat freely. Foods eaten at lunch were weighed and energy intake calculated from nutrient composition data. Appetite was assessed at baseline and at 30-min intervals between meals by line ratings of hunger, fullness, nausea and how much could be eaten. The optimum time for correlations both among appetite ratings and between appetite and lunch intake was just before the lunch. Mean coefficients of repeatability (21-38 mm) and correlation coefficients (0.67-0.71) at that point in time were similar to those reported previously in young adults. Thus, in older and well as young adults, the size of a meal is most closely related to rated appetite just before the meal. PMID- 15527925 TI - Sweet preferences and sugar consumption of 4- and 5-year-old children: role of parents. AB - We investigated the relationships in children between rules that restrict consumption of mono- and disaccharides (MDS), consumption of MDS and preferences for sucrose-containing orangeade. The background ideas of restriction rules we also investigated. To this end, 44 children (5.1+/-0.5 years) performed a rank order and paired-comparison test of preference for five orangeades, which differed in sucrose concentration (0.14, 0.20, 0.29, 0.42, 0.61 M sucrose). Parents filled out a questionnaire concerning restriction rules and their children's consumption of MDS-containing foods. Stronger restriction rules were related to a lower consumption of beverages that contained MDS and to a lower consumption of MDS-containing foods during breakfast and lunch. The most freedom to choose foods that contain MDS was given during the afternoon. Fifty-five percent of the children who were highly restricted showed a preference for the highest concentration of sucrose in orangeade. None of these children preferred the orangeade with the lowest concentration of sucrose. While 19% of the children who were little restricted preferred the beverage with the lowest concentration of sucrose, 33% preferred the beverage with the highest concentration. These parents generally believed that sugar has a bad effect on health and had similar background ideas concerning restriction rules. PMID- 15527926 TI - The relationship between parental factors at infancy and parent-reported control over children's eating at age 7. AB - The objective of this study was to determine predictors of parental control over children's eating. Data were obtained from 135 children and their parents from the Stanford Infant Growth Study. Assessments were obtained from parents at their child's birth and age 7. Parental body dissatisfaction at their child's birth predicted pressure to eat for both sons and daughters. In addition, minority status, parent being born outside of the United States, and a mother's early return to work predicted parental pressure to eat for sons. For daughters, an early maternal pushy feeding style also predicted parental pressure to eat. Parental hunger, a parental history of eating disorders and parent being born outside of the United States predicted food restriction for daughters. There were no predictors of food restriction for sons. We concluded that parental control appears not always to be simply a reaction to a child's lack of self-control or overweight, but may be present before such issues arise, particularly with the presence of certain cultural factors. PMID- 15527927 TI - Effects of short-term overfeeding on hunger, satiety, and energy intake in thin and reduced-obese individuals. AB - BACKGROUND: Certain individuals appear to be resistant to weight gain in an obesigenic environment, yet the mechanisms for this adaptation are unclear. These individuals may sense positive energy balance more appropriately than those individuals prone to weight gain. RESEARCH METHODS AND PROCEDURES: Thirteen thin (7 women, 6 men) and 9 reduced-obese (5 women, 4 men) individuals were studied. Measures of intake were obtained using visual analog scales before and after each meal during eucaloric feeding and during three days of 50% overfeeding. Ad libitum energy intake was measured post-overfeeding. RESULTS: Overfeeding resulted in a significant reduction in pre-meal hunger in the thin (68+/-6 to 41+/-6 mm, p<0.0001) compared to the reduced-obese individuals (63+/-7 to 65+/-7 mm, p=0.67). There was a significantly greater increase (p=0.0016) in post-meal satiety scores during overfeeding in the thin (65+/-4 to 88+/-4 mm, p<0.0001) compared to the reduced-obese individuals (72+/-5 to 80+/-5 mm, p=0.04). Thin women had a 30% reduction in daily energy intake (1831+/-47 to 1273+/-79 kcal/day, p=0.005) after overfeeding while no difference was seen in the thin men or reduced-obese group. DISCUSSION: Thin individuals appear to sense positive energy balance more appropriately with changes in measures of intake than reduced obese individuals. These findings are especially true in thin women. PMID- 15527928 TI - Long lasting effects of rearing by an ethanol-consuming dam on voluntary ethanol consumption by rats. AB - For exposure to alcohol early in life to potentiate alcohol abuse in adolescence or adulthood, consequences of early exposure to alcohol must be of considerable duration. In two experiments using Norway rats as subjects, we examined effects of exposure during weaning to a dam consuming ethanol on adolescents' later affinity for ethanol. In a preliminary experiment, we offered rat pups a choice between 8% ethanol and water for 7 days immediately after they were weaned at 26 days of age. Pups whose dam had ingested 8% ethanol for 6 days either immediately or 1 week before we weaned them drank more ethanol than pups whose dam drank only water during the same period. Independent groups of rats reared by a dam consuming 8% ethanol from postnatal days 18 to 26 and tested 1, 2, 4 or 6 weeks later all drank significantly more 8% ethanol at testing than did pups whose dam drank only water. Our data also provided confirmation of previous reports of an experience-independent greater affinity for ethanol in younger rats. PMID- 15527929 TI - Old and alone: barriers to healthy eating in older men living on their own. AB - Ageing is associated with reduced energy intake and loss of appetite. Older men tend to have poorer dietary intakes including consumption of fewer fruits and vegetables in comparison to older women. Living and eating alone further diminishes food consumption and dietary quality. The aim of the present study was to explore food choice and energy intake in older men living alone using both quantitative and qualitative methods. 39 older men were interviewed and completed questionnaires on health, food choice, dietary patterns and appetite. Few men managed to consume recommended levels of energy, essential trace elements or vitamins A and D. Age and BMI failed to predict patterns of intake, but men with good cooking skills reported better physical health and higher intake of vegetables. However, cooking skills were negatively correlated with energy intake. Men who managed to consume at least 4 portions of fruits and vegetables each day had significantly higher vitamin C levels, a greater percentage of energy as protein and generally more adequate diets. Interviews revealed that poor cooking skills and low motivation to change eating habits may constitute barriers to improving energy intake, healthy eating and appetite in older men (193). PMID- 15527930 TI - Patterns of eating and movement that best maintain reduction in overweight. AB - Two studies of the influences of specific patterns of eating and exercising behaviour on body weight in English Midlands women were re-analysed using correlations as the measure of effect size. As predicted from computational modelling of hunger-sating mechanisms, avoiding energy-containing drinks and foods at the ends of and between meals was the behaviour most influencing year long weight loss. However, although eating between meals is often called snacking, the term 'snack' appeared to be too ambiguous in this culture for its use in helping efforts to control weight. Avoidance of particular sorts of fat rich foods was also associated with longer-term weight loss. Attempts at severe restriction of intake at mealtimes were associated with weight loss during a period of intensive dieting, but did not contribute to maintenance of that weight reduction. Using diet formulae to attain rapid weight loss was associated with significant weight gain over a year. These results support the suggestion that the first line of defence against weight gain is avoiding all sources of energy during drink breaks, with personally relevant advice on lower fat versions of particular foods also being important. Continued neglect of the behaviour specific correlational approach to gaining evidence for less fattening habits does nothing to slow the rise in obesity. PMID- 15527931 TI - Craving and startle modification during in vivo exposure to food cues. AB - RATIONALE AND OBJECTIVES: Food craving and reactivity to food cues are related to food intake. Research has shown that both startle magnitude and prepulse inhibition of startle may be useful indices of cue reactivity and drug craving. The goal of this study was to assess the utility of these paradigms for the study of food craving in food-deprived participants. METHODS: Twenty-nine non-smoking, food-deprived undergraduate participants were exposed to a counterbalanced series of food, cigarette, and neutral cues. During each 4.5-min cue, nine startle probes (102-dB white noise) were presented, and a prepulse (20-ms noise 8 dB above background) preceded 2/3 of the probes by 60 or 120 ms. Subjects were divided into those high and low in food-craving on the basis of subjective craving to food cues. Responses to food cues were compared to smoking cues to control for attention and arousal. RESULTS: Both probe-alone startle magnitude and percent prepulse inhibition were significantly reduced during the food cue compared to the smoking cue among high food cravers, but not low food cravers. These processes seemed to be independent, as cue-induced changes in prepulse inhibition were uncorrelated with cue effects on probe-alone startle magnitude. CONCLUSIONS: Both probe-alone startle magnitude and prepulse inhibition are sensitive to cue-induced craving for food, but appear to assess separate processes. Startle methodologies may prove useful in understanding basic processes that regulate food intake. PMID- 15527932 TI - Gender differences in factors affecting rejection of food in healthy young Swedish adults. AB - With the objectives to better understand gender-related differences in variables of importance for food intake, and interrelations between these variables, 100 healthy, young women and 100 healthy, young men responded to self-administrated questionnaires about general food rejection, learned illness-associated food aversions, disgust (the Disgust Scale), food neophobia (the Food Neophobia Scale), nausea and appetite. The results show that food rejection and aversions were more common in women (69 and 38%, respectively) than in men (47 and 18%), and that women are more disgust sensitive than men. However, no differences between women and men were observed regarding reasons for rejecting food (predominantly sensory attributes), prevalence of gastrointestinal illness as an associated aversion symptom (95 vs 89%), type of aversive food due to associated illness (predominantly high protein items), or food neophobia. Based on path analyses, a model is proposed of interrelations between disgust, food neophobia, learned food aversions, nausea, appetite, and general food rejection in healthy young adults. PMID- 15527933 TI - Impulsivity and test meal intake in obese binge eating women. AB - Greater impulsivity has been observed in those with chemical (cocaine, marijuana, alcohol) and behavioral addictions (gambling, sex, shopping), as well as in individuals with personality and conduct disorders. Greater impulsivity has also been described in those with Bulimia Nervosa and attributed to aberrations in serotonin, as has eating in response to negative affect. However, less is known about the impact of impulsivity on eating behavior in obese humans in general, and in those who meet sub-clinical and full clinical criteria for Binge Eating Disorder (BED) in particular. Using a laboratory test meal paradigm, we demonstrated: (1) greater Motor Impulsivity (Barratt Impulsivity Scale (BIS) (p = 0.05) in those with BED (n = 11) as compared to those without BED (n = 11) (2) a positive correlation between BED criteria and BIS scores (p < 0.01) (3) a positive correlation between test meal duration and Zung Depression Score, and (4) a positive correlation between Motor Impulsivity and mood rated before consuming the test meal. These associations suggest potential aberrations in serotonin transmission in BED, and a possible target for pharmacotherapy of BED especially in those who are resistant to Cognitive Behavioral Therapy. PMID- 15527934 TI - Consumer perceptions of food safety risk, control and responsibility. AB - Consumer perceptions of risk, control and responsibility related to microbial food safety in domestic food preparation have been investigated. Results showed that consumers demonstrated judgements of 'optimistic-bias' and the 'illusion of control', as well as notions of perceived invulnerability to food poisoning from self-prepared foods. Statistical associations between perceptions of personal risk, control and responsibility and risk and control attributed to 'other people' have been identified. Findings may have negative implications for the effectiveness of consumer food safety education initiatives. It is suggested that consideration of such judgements and associations during the development of future consumer food safety risk communication strategies may increase their effectiveness. PMID- 15527935 TI - Neural control of appetite: cross-talk between homeostatic and non-homeostatic systems. AB - The new lifestyle in the modern world is causing a rapid increase in the prevalence of obesity and associated health problems. Increased availability of palatable and energy dense foods, combined with a lack of physical activity overpower a homeostatic regulatory system that evolved to survive periods of famine rather than preventing obesity. Environmental and lifestyle factors influence energy balance mainly through their impact on cortico-limbic brain structures dealing with reward, cognitive, and social aspects of food intake and voluntary physical activity. To find new behavioral and pharmacological treatments of obesity, it will be important to identify the specific pathways that link these externally driven processes with the homeostatic regulatory system. PMID- 15527936 TI - The effect of information about fat content on food consumption in overweight/obese and lean people. AB - This study investigated how fat content labels (high-fat vs. low-fat) influence milkshake consumption in obese/overweight people (n = 23) as compared to lean people (n = 21). Participants 'tasted' two isocaloric milkshakes in a staged taste test on two occasions. On one occasion the milkshakes were labeled high fat, whereas on the other occasion they were labeled low-fat. The label-effect was in the expected direction of less (estimated future) consumption in the high fat label condition, but was not significant for the current consumption and not different between groups. Unexpectedly, order (label high-fat first vs. label low fat first) had a large effect on consumption. PMID- 15527937 TI - A comparison of energy intake between eating positions in a NHS hospital--a pilot study. AB - Malnutrition and the under-consumption of food in hospitals is prevalent and in UK hospitals, the consumption of meals is mainly a solitude event, despite evidence to show that eating in the presence of others can actually increase food intake. Dietary data were collected for three consecutive 24 h periods (n=13) from patients who consumed their meals either in bed, at the side of the bed or in the presence of others. Results show a significant increase (p<0.05) in the mean daily energy intake for those sitting around a table in the presence of others. Although a small pilot study, the results confirm the value of social facilitation in improving the under-consumption of food when in hospital. PMID- 15527938 TI - Shared lives-an opportunity for obesity prevention? AB - A longitudinal design was used to survey individual members of 22 couples about food choices, dietary intake and body weight around three months prior to cohabitation and again about three months after the moving-in date. Changes after cohabitation included an increase in shared meal occasions, increased likelihood of including alcohol at meal times and social support for dietary temptations (and restrictions). Body weight increased significantly in women (58.3+/-7.1 - 59.8+/-7.9 kg) and in men (76.7+/-12.0 - 78.4+/-12.5 kg). This life stage may be a missed opportunity for obesity prevention initiatives. PMID- 15527939 TI - The relationship between susceptibility to nausea and vomiting and the possession of conditioned food aversions. AB - Because nausea and vomiting play an important role in the formation of conditioned food aversions, individual differences in susceptibility to nausea and vomiting may contribute to interindividual variation in conditioned food aversion acquisition. Two surveys reveal that, among female participants, self reported motion sickness susceptibility is positively correlated with the number of conditioned food aversions possessed, but no such correlation appears among male participants. PMID- 15527940 TI - Is there a "molecular Nirvana Principle"? Towards a unified resolutional model of the biological symbol-matter dichotomy. AB - In the present paper, the metapsychological "Nirvana Principle" is investigated evolutionarily at the earliest forms of life in a highly tentative way. A corresponding "molecular Nirvana Principle" is proposed, where the recent suggestions of the "internal measurement" biophysical quantum-molecular research programme of modern quantum biology are introduced, in relation to the former metapsychological theory, conceived to be valid in the entire realm of living systems (just as it was intended by the original author). By an appropriate introduction of a special primordeal dynamical time inversion symmetry breaking, originating in a premeval self-measurement in a composite nucleic acid-protein system, a special internal symmetry restoration time series is defined. In this way, a strictly physically defined self-identity ("molecular Nirvana," special physical symmetry restored) is derived, which is put equal to the quantum physical equivalent and root of the goals of evolutionarily higher level fundamental drives (the "Nirvana Principle"). It is shown that it is a natural requirement that the following internal regressive time (-reversal) physical molecular relations (and so the ultimate time symmetry) is mapped onto space, as is also suggested by some symbol-theoretical propositions. PMID- 15527941 TI - An algorithm for the study of DNA sequence evolution based on the genetic code. AB - Recent studies of the quantum-mechanical processes in the DNA molecule have seriously challenged the principle that mutations occur randomly. The proton tunneling mechanism causes tautomeric transitions in base pairs resulting in mutations during DNA replication. The meticulous study of the quantum-mechanical phenomena in DNA may reveal that the process of mutagenesis is not completely random. We are still far away from a complete quantum-mechanical model of DNA sequence mutagenesis because of the complexity of the processes and the complex three-dimensional structure of the molecule. In this paper we have developed a quantum-mechanical description of DNA evolution and, following its outline, we have constructed a classical model for DNA evolution assuming that some aspects of the quantum-mechanical processes have influenced the determination of the genetic code. Conversely, our model assumes that the genetic code provides information about the quantum-mechanical mechanisms of mutagenesis, as the current code is the product of an evolutionary process that tries to minimize the spurious consequences of mutagenesis. Based on this model we develop an algorithm that can be used to study the accumulation of mutations in a DNA sequence. The algorithm has a user-friendly interface and the user can change key parameters in order to study relevant hypotheses. PMID- 15527942 TI - Use of chemically modified PMMA microspheres for enzyme immobilization. AB - Modified poly(methyl methacrylate) (PMMA) microspheres, about 7microm in diameter, carrying aldehyde groups on their surfaces were synthesized and used as the support for enzyme immobilization. The immobilizing behavior as well as the properties of immobilized enzyme was studied. The amount of bound enzyme can be extended to 76.8mg g(-1) support, which is relatively much higher than other supports. The kinetic investigation derived from three typical models shows that the practical process is more complicated than the ideal condition, with one or more interactions being involved in the immobilization process. The K(m) value is actually larger and V(max) is smaller in the immobilized form than those in the free form. The increased resistance of the immobilized enzyme against the changes of temperature indicates that immobilizing enzyme onto the modified microspheres is useful for enzyme immobilization. PMID- 15527943 TI - Computational identification and sequence analysis of stop codon readthrough genes in Oryza sativa. AB - Using an approach based on the Readthrough Candidate Extraction System (RCES), we extracted 111 candidates from 9620 gene sequences of rice. The results of homology search and sequence analysis demonstrated that these candidates included actual readthrough genes that would be important for further investigating the mechanism of translation termination regulated by readthrough event, and could also give some useful clues for functional genome annotation. Between the candidates and non-candidates of gene sequences in rice, there exist significant base biases at the positions surrounding the stop codons. These positions, especially both -1 and +4, are referred to as part of an extended stop signal. In candidates, G at position -1, and G or C at position +4 are much more favored than that in non-candidates. Both stop sequence patterns, GUAGC and GUGAG, might drive high readthrough efficiency in rice. Secondary structure analysis revealed that the -1 and +1 amino acids around the first stop codon of candidates have a strong bias toward arginine, particularly the +1 position (20.7%), which indicated that the amino acids at the readthrough region being frequently located in the hydrophilic region of beta-turn might be a determinant for efficient translation termination or not. PMID- 15527944 TI - Circadian rhythm formation in plant seedling: global synchronization and bifurcation as a coupled nonlinear oscillator system. AB - Circadian rhythm formation is studied in seedlings after germination measuring their respiratory metabolism. The circadian rhythm is clearly observed at about 170h (the onset time t(CR-ON)) after germination of seeds in natural conditions in a dark incubator. There are no clear cyclic signals in gas exchange before t(CR-ON). Application of external triggers (temperature shocks) near the onset of the rhythm in seedling growth strongly affects formation processes. The onset is shifted earlier up to 50h by application of perturbations. This fact may suggest that the circadian rhythms appear via subcritical bifurcation. PMID- 15527945 TI - Quantum computation, non-demolition measurements, and reflective control in living systems. AB - Internal computation underlies robust non-equilibrium living process. The smallest details of living systems are molecular devices that realize non demolition quantum measurements. These smaller devices form larger devices (macromolecular complexes), up to living body. The quantum device possesses its own potential internal quantum state (IQS), which is maintained for a prolonged time via reflective error-correction. Decoherence-free IQS can exhibit itself by a creative generation of iteration limits in the real world. It resembles the properties of a quasi-particle, which interacts with the surround, applying decoherence commands to it. In this framework, enzymes are molecular automata of the extremal quantum computer, the set of which maintains highly ordered robust coherent state, and genome represents a concatenation of error-correcting codes into a single reflective set. The biological evolution can be viewed as a functional evolution of measurement constraints in which limits of iteration are established, possessing criteria of perfection and having selective values. PMID- 15527946 TI - Interactions between learning and evolution: the outstanding strategy generated by the Baldwin effect. AB - The Baldwin effect is known as an possible interaction between learning and evolution, where individual lifetime learning can influence the course of evolution without using any Lamarckian mechanism. Our concern is to consider the Baldwin effect in dynamic environments, especially when there is no explicit optimal solution through generations and this solution depends only on interactions among agents. We adopted the iterated Prisoner's Dilemma as a dynamic environment, introduced phenotypic plasticity into its strategies, and conducted computational experiments, in which phenotypic plasticity is allowed to evolve. The Baldwin effect was observed in the experiments as follows: First, strategies with enough plasticity spread, which caused a shift from defect oriented populations to cooperative populations. Second, these strategies were replaced by a strategy with a modest amount of plasticity generated by interactions between learning and evolution. By making three kinds of analysis, we have shown that this strategy provides outstanding performance in comparison with other deterministic strategies. Further experiments towards open-ended evolution have also been conducted so as to generalize our results. PMID- 15527948 TI - A new DNA computing model for the NAND gate based on induced hairpin formation. AB - Hairpin structure of DNA molecules has been widely employed in a variety of biosensors and in nanoscale molecular assembly applications. For example, the well known molecular beacons can report the presence of specific nucleic acids in homogeneous solutions with high accuracy. Recently, Smith et al. proposed the induction of hairpin formation through sequence-specific binding of a small molecule ligand to a G-G mismatch. Not only did this method offer great flexibility in controlling hairpin formation, more importantly the induced hairpin maintains high degree of sensitivity toward specific hybridization. In this contribution, we present a theoretical model for the logical NAND gate based on induced hairpin formation. PMID- 15527947 TI - Tubulin dipole moment, dielectric constant and quantum behavior: computer simulations, experimental results and suggestions. AB - We used computer simulation to calculate the electric dipole moments of the alpha and beta-tubulin monomers and dimer and found those to be |p(alpha)| = 552D, |p(beta)| = 1193D and |p(alphabeta)| = 1740D, respectively. Independent surface plasmon resonance (SPR) and refractometry measurements of the high-frequency dielectric constant and polarizability strongly corroborated our previous SPR derived results, giving Deltan/Deltac approximately 1.800 x 10(-3)ml/mg. The refractive index of tubulin was measured to be n(tub) approximately 2.90 and the high-frequency tubulin dielectric constant k(tub) approximately 8.41, while the high-frequency polarizability was found to be alpha(tub) approximately 2.1 x 10( 33)C m(2)/V. Methods for the experimental determination of the low-frequency p are explored, as well as ways to test the often conjectured quantum coherence and entanglement properties of tubulin. Biobits, bioqubits and other applications to bioelectronics are discussed. PMID- 15527950 TI - A parameter control method in reinforcement learning to rapidly follow unexpected environmental changes. AB - In order to rapidly follow unexpected environmental changes, we propose a parameter control method in reinforcement learning that changes each of learning parameters in appropriate directions. We determine each appropriate direction on the basis of relationships between behaviors and neuromodulators by considering an emergency as a key word. Computer experiments show that the agents using our proposed method could rapidly respond to unexpected environmental changes, not depending on either two reinforcement learning algorithms (Q-learning and actor critic (AC) architecture) or two learning problems (discontinuous and continuous state-action problems). PMID- 15527949 TI - Adaptive chronic infection, structured stress, and medical magic bullets: do reductionist cures select for holistic diseases? AB - With a generalized language-of-thought argument for immune cognition, we model how population-directed, structured, psychosocial stress can impose an image of itself on the coevolutionary conflict between a highly adaptive chronic infection and the immune response. As population-level structured stress appears a fundamental part of the biology of disease, we raise the possibility that simplistic individual-oriented magic-bullet drug treatments, vaccines, and risk reduction programs that do not address the fundamental living and working conditions which underlie disease ecology will fail to control many current epidemics. In addition, such reductionist interventions may go so far as to select for more holistic pathogens characterized by processes operating at multiple levels of biocultural organization. The complications are representative of the concerns of cultural immunology, a new field of study. PMID- 15527951 TI - A new theory of the origin of cancer: quantum coherent entanglement, centrioles, mitosis, and differentiation. AB - Malignant cells are characterized by abnormal segregation of chromosomes during mitosis ("aneuploidy"), generally considered a result of malignancy originating in genetic mutations. However, recent evidence supports a century-old concept that maldistribution of chromosomes (and resultant genomic instability) due to abnormalities in mitosis itself is the primary cause of malignancy rather than a mere byproduct. In normal mitosis chromosomes replicate into sister chromatids which are then precisely separated and transported into mirror-like sets by structural protein assemblies called mitotic spindles and centrioles, both composed of microtubules. The elegant yet poorly understood ballet-like movements and geometric organization occurring in mitosis have suggested guidance by some type of organizing field, however neither electromagnetic nor chemical gradient fields have been demonstrated or shown to be sufficient. It is proposed here that normal mirror-like mitosis is organized by quantum coherence and quantum entanglement among microtubule-based centrioles and mitotic spindles which ensure precise, complementary duplication of daughter cell genomes and recognition of daughter cell boundaries. Evidence and theory supporting organized quantum states in cytoplasm/nucleoplasm (and quantum optical properties of centrioles in particular) at physiological temperature are presented. Impairment of quantum coherence and/or entanglement among microtubule-based mitotic spindles and centrioles can result in abnormal distribution of chromosomes, abnormal differentiation and uncontrolled growth, and account for all aspects of malignancy. New approaches to cancer therapy and stem cell production are suggested via non-thermal laser-mediated effects aimed at quantum optical states of centrioles. PMID- 15527952 TI - History dependent effects on phenotypic expression of a newly emerged gene. AB - In this study, we investigate the history dependence of the penetrance of a newly emerged gene. Penetrance is defined as the percentage of individuals with a given genotype who exhibit the phenotype associated with that particular genotype. Here, we used the glutamine synthetase gene and its mutants with lower fitness as model genes. They were introduced into host cells of Escherichia coli deprived of the gene, and their penetrance was measured using the host having a different history: either with or without glutamine starvation. Results show that for all genes tested, the value of penetrance was higher when they were introduced into the host cell without starvation than that when introduced into the starved cell, demonstrating the history dependence of the penetrance of a newly emerged gene. In addition, genes with lower fitness showed lower penetrance, and the effect of the difference in fitness on gene penetrance also depended on the history of the host cell. PMID- 15527953 TI - Stochastic kinetics of reproduction of virions inside a cell. AB - We analyze intracellular viral kinetics in the framework of the model incorporating viral genome replication, mRNA synthesis and degradation, protein synthesis and degradation, capsid assembly, and virion release from a cell. Due to the existence of the critical concentration of viral capsid proteins and other features of reproduction of virions inside a cell, the kinetics is demonstrated to exhibit three distinct initial stages. Specifically, (i) the exponential growth of the viral genome, mRNA and protein concentrations is followed by (ii) the transient stage to (iii) the steady-state regime. The formation of mature virions starts during the transient stage. Comparison of the kinetics, obtained by using the mass-action law and Monte Carlo (MC) technique, indicates that they are nearly identical during the initial exponential growth of the viral intermediates and also during the steady-state stage. The transition from the initial stage to the steady-state regime occurs however somewhat faster in the determenistic case even if the steady-state populations of virions and genomes are appreciable (e.g., about 250 and 500, respectively). PMID- 15527954 TI - Artificial life simulation of self-assembly in bacteriophage by movable finite automata. AB - This paper presents a model which is based on biological research using the movable finite automata (MFA) on a self-assembly of T4 phage, and exhibits the results of artificial life simulation. In the previous work, Thompson and Goel [Artificial Life, Addison Weley, 1989, pp. 317-340; Biosystems 18 (1985) 23; J. Theor. Biol. 131 (1988) 351] presented the movable finite automata (MFA) which has a capability of moving on finite automata, and simulated on a computer. They were represented individual rectangular boxes, however, the results of simulation was different from real T4 phage. We propose the sphere model as a protein structure, and simulate the self-assembly of the entire structure of the T4 phage on a computer. PMID- 15527955 TI - On the optimality of the genetic code, with the consideration of termination codons. AB - The existence of nonrandom patterns in codon assignments is supported by many statistical and biochemical studies. The canonical genetic code is known to be highly efficient in minimizing the effects of mistranslation errors and point mutations. For example, it is known that when an error induces the conversion of an amino acid to another, the biochemical properties of the resulting amino acid are usually very similar to that of the original. Prior studies include many attempts at quantitative estimation of the fraction of randomly generated codes which, based upon load minimization, score higher than the canonical genetic code. In this study, we took into consideration both the relative frequencies of amino acids and nonsense mistranslations, factors which had been previously ignored. Incorporation of these parameters, resulted in a fitness function (phi) which rendered the canonical genetic code to be highly optimized with respect to load minimization. Considering termination codons, we applied a biosynthetic version of the coevolution theory, however, with low significance. We employed a revised cost for the precursor-product pairs of amino acids and showed that the significance of this approach depends on the cost measure matrix used by the researcher. Thus, we have compared the two prominent matrices, point accepted mutations 74-100 (PAM(74-100)) and mutation matrix in our study. PMID- 15527956 TI - Bio-steps beyond Turing. AB - Are there 'biologically computing agents' capable to compute Turing uncomputable functions? It is perhaps tempting to dismiss this question with a negative answer. Quite the opposite, for the first time in the literature on molecular computing we contend that the answer is not theoretically negative. Our results will be formulated in the language of membrane computing (P systems). Some mathematical results presented here are interesting in themselves. In contrast with most speed-up methods which are based on non-determinism, our results rest upon some universality results proved for deterministic P systems. These results will be used for building "accelerated P systems". In contrast with the case of Turing machines, acceleration is a part of the hardware (not a quality of the environment) and it is realised either by decreasing the size of "reactors" or by speeding-up the communication channels. Consequently, two acceleration postulates of biological inspiration are introduced; each of them poses specific questions to biology. Finally, in a more speculative part of the paper, we will deal with Turing non-computability activity of the brain and possible forms of (extraterrestrial) intelligence. PMID- 15527957 TI - Cluster and information entropy patterns in immunoglobulin complementarity determining regions. AB - Previous studies of antibody binding domains have established many crucial features that include important structural positions, canonical formations, and the geometric correlations with the binding site nature and topography. In this work, position-specific frequency and hierarchical clustering analysis are used to explore the statistical pattern of the residues in the complementarity determining regions of human antibodies. In addition, Shannon's information entropy is computed for the entire heavy and light chains and compared with germline patterns to seek variability due to antibody clonal selection. Results are compared with reported analyses based on structural data and ligand-protein contact point computations based on Protein Data Bank records. Observations derived from the present sequence analysis are consistent with previous structural based methods. In the absence of structural data, methods used in this work can be effective and efficient computational tools used for identifying residues that are important for antigen targeting and predicting the probable amino acid distribution expected at these positions. The results in turn can be applied to help design or plan mutagenesis experiments to improve the binding properties of antibodies. PMID- 15527958 TI - Ecosystem growth and development. AB - One of the most important features of biosystems is how they are able to maintain local order (low entropy) within their system boundaries. At the ecosystem scale, this organization can be observed in the thermodynamic parameters that describe it, such that these parameters can be used to track ecosystem growth and development during succession. Thermodynamically, ecosystem growth is the increase of energy throughflow and stored biomass, and ecosystem development is the internal reorganization of these energy mass stores, which affect transfers, transformations, and time lags within the system. Several proposed hypotheses describe thermodynamically the orientation or natural tendency that ecosystems follow during succession, and here, we consider five: minimize specific entropy production, maximize dissipation, maximize exergy storage (includes biomass and information), maximize energy throughflow, and maximize retention time. These thermodynamic orientors were previously all shown to occur to some degree during succession, and here we present a refinement by observing them during different stages of succession. We view ecosystem succession as a series of four growth and development stages: boundary, structural, network, and informational. We demonstrate how each of these ecological thermodynamic orientors behaves during the different growth and development stages, and show that while all apply during some stages only maximizing energy throughflow and maximizing exergy storage are applicable during all four stages. Therefore, we conclude that the movement away from thermodynamic equilibrium, and the subsequent increase in organization during ecosystem growth and development, is a result of system components and configurations that maximize the flux of useful energy and the amount of stored exergy. Empirical data and theoretical models support these conclusions. PMID- 15527959 TI - Drosophila, an emerging model for cardiac disease. AB - A variety of studies that are currently underway may validate the fruit fly as an in vivo model for analyzing genes involved in cardiac function. Many mutations in conserved genetic pathways have been found, including those controlling development and physiology. Because homologous genes control early developmental events as well as functional components of the Drosophila and vertebrate hearts, the fly is the simplest existing model system that can be used to assay genes involved in human congenital heart disease (CHD). The wide variety of genetic tools available to Drosophila researchers offers many technical advantages for rapidly screening through large numbers of candidate genes. Thus, an important future and long-term direction is likely to be the use of Drosophila as a vehicle for analyzing polygenic traits as an aid in human genetics. One can anticipate a time in the not too distant future when mutant lines exist for every gene in vertebrate systems, such as mice and zebrafish. However, one of the enduring problems that will not easily be addressed by such resources will be the tracking of complex traits defined by polygenic variants. For this level of genetic analysis, simple genetic model systems including yeast, Caenorhabditis elegans, and Drosophila melanogaster will undoubtedly play a crucial ongoing role. Of them, Drosophila will be critical for examining gene networks involved in organogenesis and is clearly the system of choice for studying cardiac development, function and aging, since among the simple genetic models it is the only one with a fluid pumping heart. PMID- 15527960 TI - Skeletal hormones and the C/EBP and Runx transcription factors: interactions that integrate and redefine gene expression. AB - Systemic hormones and local growth factors have significant and often complex roles in normal tissue development, growth, remodeling, and repair. Early efforts in skeletal tissue attempted to define active panels of these agents and their direct effects on cell proliferation, matrix production, and secretion of other soluble mediators of differentiated cell function. Initial results resolved many of these questions and began to unveil functional interactions between specific hormones and growth factors. More recent evidence suggests that interactions between individual hormone systems also occur in less anticipated but probably not less meaningful ways. In some cases, these interactions may help to define a spectrum of effects on gene expression by focusing, refocusing, or integrating the activity of previously recognized transcription regulators. Other studies in isolated osteoblasts predict that certain steroid hormones have distinctive effects on specific transcription factors with important roles in bone growth and repair. In this review, we focus on studies that define functional and physical interactions between molecular mediators of hormone activity that could directly effect skeletal growth factor biology. PMID- 15527961 TI - Intron-derived microRNAs--fine tuning of gene functions. AB - New evidence suggests that abundant small RNAs, microRNAs (miRNAs) act as regulatory vehicles to repress translation or cleave RNA transcripts, depending on their complementarity to the target gene, and result in modulation of gene expression. This review describes that the miRNAs derived from introns can suppress intracellular RNA homologues and regulate the gene function. PMID- 15527962 TI - Characterization of synectin expression and promoter activity. AB - Synectin, a PDZ2 protein, binds to a number of plasma membrane receptors. We characterized synectin promoter and carried out a detailed expression survey. The protein is widely distributed in adult cells and tissues. Expression in the mouse embryo is apparent at day 4.5, peaks at day 12.5 and declines to its adult levels by day 18.5. The level of synectin transcription was minimally affected by exposure to cytokines, growth factors or hypoxia. In summary, synectin's early appearance, constant nature, and ubiquitous distribution suggest that it may be a key structural protein involved in mediation of outside-in signaling. PMID- 15527963 TI - Functional characterization of a human histone gene cluster duplication. AB - Histones are the major protein component of nucleosomes, and de novo histone synthesis is essential for packaging newly replicated DNA into chromatin. As a result, histone gene expression is exquisitely and functionally coupled with DNA replication. Vastly divergent organisms such as yeast, fly and human all demonstrate the phylogenetically conserved propensity to maintain clustering of histone genes at one or more genomic loci. Although specific mechanisms are unclear, clustering is presumed to be important for common stringent transcriptional control of these genes at the G1/S phase transition. In this study, we describe a genomic duplication of the human histone gene cluster located at chromosome 1q21, which effectively doubles the previously known size and gene number of that cluster. The duplication persists in all examined tissues and cell lines, and the duplicated genes are transcriptionally active. Levels of messenger RNAs for duplicated histone H4 genes are high relative to those for non duplicated H4 genes. Our data suggest that transcriptionally robust histone H4 genes may have been preferentially duplicated during evolution. PMID- 15527964 TI - Identification of Xenopus cyclin-dependent kinase inhibitors, p16Xic2 and p17Xic3. AB - The Cip/Kip family of mammalian cyclin-dependent kinase (cdk) inhibitors plays important roles in development, particularly in cell fate determination and differentiation, in addition to their function of blocking cell cycle progression. We have identified two novel members of the Kip/Cip cdk inhibitor family, p16Xic2 and p17Xic3, from Xenopus laevis. Sequence analysis revealed that p16Xic2 and p17Xic3 are orthologues of mammalian p21Cip1 and p27Kip1, respectively. Overexpression of these inhibitors results in cell cycle arrest by inhibition of cdk2 activity. Interestingly, the expression of these inhibitors is highly developmentally regulated. p16Xic2 is highly expressed in differentiating somite, tail bud, lens, and cement gland, while p17Xic3 is expressed in the central nervous system. In a retinal cell fate determination assay, both p16Xic2 and p17Xic3 have an activity that influences cell fate determination. These observations suggest that p16Xic2 and p17Xic3 might be involved in cell fate determination in a tissue-specific manner by coordinating proliferation and differentiation as observed with p27Xic1. PMID- 15527965 TI - Identification and characterization of a novel Drosophila melanogaster glutathione S-transferase-containing FLYWCH zinc finger protein. AB - Glutathione SH-transferase (GST) is a 25-kDa protein and a member of a large family that plays a critical role in the cellular homeostasis of all organisms. In this report, we describe a novel GST-containing protein identified and cloned from Drosophila. This 1045 amino acid protein possesses a zinc finger domain with a tandem array of four FLYWCH zinc finger motifs at its N-terminus and a C terminal domain that shares a 46% homology with GST. The gene maps to chromosome 3 at position 84C6. Further characterization of this protein shows that it localizes to the cytoplasm of fly cells and is expressed through all stages of fly embryonic development. It binds to glutathione-S agarose beads in vitro. These results indicate that this new protein belongs to the GST family, thus named a Drosophila GST-containing FLYWCH zinc finger protein (dGFZF). PMID- 15527966 TI - Characterization of Oryza sativa telomerase reverse transcriptase and possible role of its phosphorylation in the control of telomerase activity. AB - Telomerase reverse transcriptase (TERT) has been characterized in the dicotyledon Arabidopsis thaliana. A TERT homolog has now been identified in the monocotyledon rice (Oryza sativa L.) on the basis of its predicted homology to the A. thaliana enzyme (AtTERT). At least five alternatively spliced transcripts of the rice TERT (OsTERT) gene were detected. The full-length OsTERT protein shares structural features with TERTs of other species, including a calculated molecular size of 144 kDa, an isoelectric point of 9.6, and conserved sequence motifs. Phylogenetic analysis showed that OsTERT clusters with AtTERT and is more related to the human and mouse enzymes than to those of yeast and ciliated protozoa, consistent with the evolutionary relations among these eukaryotes. Telomerase activity was abundant in shoot apices and cultured cells but was low or absent in leaves or roots of rice plants, whereas similarly spliced OsTERT transcripts were detected in all tissues examined and cultured cells. Similar to mouse and human TERT proteins, OsTERT contains two putative phosphorylation sites for Akt kinase. Incubation of a rice cell extract with Akt or with protein phosphatase 2A potentiated or inhibited telomerase activity, respectively, whereas Akt did not affect the activity in Arabidopsis cell extract. In addition, the kinase activated the telomerase in a leaf extract. The mechanism of telomerase regulation in rice thus appears to differ from that in Arabidopsis (which is mediated predominantly at the level of AtTERT transcription), possibly reflecting the taxonomic distance between monocotyledons and dicotyledons. PMID- 15527967 TI - Differential target gene activation by TBX2 and TBX2VP16: evidence for activation domain-dependent modulation of gene target specificity. AB - The determinants of in vivo target site selectivity by transcription factors are poorly understood. To find targets for the developmentally regulated transcription factor TBX2, we generated stable transfectants of human embryonic kidney cells (293) that express a TBX2-ecdysone receptor (EcR) chimeric protein. While constitutive expression of TBX2 is toxic to 293 cells, clones expressing TBX2EcR are viable in the absence of an EcR ligand. Using cDNA arrays and quantitative PCR, we discovered nine genes whose expression was increased, but no genes whose expression was reduced, following 24 h of induction with Ponasterone A (PonA), a ligand for EcR. Since TBX2 was reported previously to be a transcriptional repressor, we also generated cell lines expressing a TBX2VP16EcR protein which we showed was a potent conditional transcriptional activator in transient transfection assays. Treatment of these cells with PonA induced the expression of five genes, none of which were affected in TBX2EcR-expressing cells. This discordance between TBX2- and TBX2VP16-regulated genes strongly suggests that specific transactivation domains can be a major determinant of gene target site selectivity by transcription factors that possess the same DNA binding domain. PMID- 15527969 TI - Multiple novel transcription initiation sites for NRG1. AB - The large neuregulin 1 gene (NRG1) has been mapped to a 1.125 Mb region on chromosome 8p11-21. Three major forms of NRG1 (types I-III), all with distinct amino-termini encoded by unique 5'-exons, have been described. We report here the discovery of nine novel NRG1 exons, including six alternative 5'-exons, increasing the number of potential promoters in NRG1 from three to nine. The novel transcripts of NRG1 described here use the novel 5'-exons which are either coding or non-coding. The functional relevance of the predicted proteins they encode has not been evaluated. Three of the novel 5'-exons are well conserved in syntenic rat and mouse sequences; they encode proteins with novel amino-termini, here termed types IV-VI. NRG1 plays a central role in neural development and is most likely involved in regulation of synaptic plasticity, or how the brain responds or adapts to the environment. The unusually complex gene structure may facilitate spatial and temporal regulation of NRG1 expression, fine-tune NRG1 protein function at different stages during development of the nervous system, and adapt responses to the environment in the adult brain. PMID- 15527968 TI - A novel gene family induced by acute inflammation in endothelial cells. AB - The aim of this study was to characterise a novel family of inflammatory genes induced by pro-inflammatory cytokines in primary human endothelial cells. Using a genome-wide array screen two previously uncharacterised genes, NLF1 and NLF2 were identified that were upregulated over 30 fold by treatment with interleukin 1beta for 2 h. They were also found to respond to tumour necrosis factor alpha, suggesting a general role in inflammation. Expression of both genes peaked 2 h after addition of interleukin 1beta, with similar kinetics to the fastest nuclear factor kappaB (NF-kappaB) induced genes. The activation of both genes by interleukin 1beta was abrogated by the proteasomal inhibitor, lactacystin which blocks activation of NF-kappaB by preventing IkappaB degradation. Furthermore, two sequences with homology to NF-kappaB binding sites in the promoter of NLF1 were found to be essential for rapid elevation in expression in response to interleukin 1beta. NLF1 and NLF2 transcripts were found predominantly in endothelial cells, and the encoded proteins were localised to the nuclear compartment suggesting a role in the regulation of transcription. Transfection of recombinant NLF into endothelial cells resulted in upregulation of the Rho kinases, Rnd1 and Gem GTPase. We propose that NLF1 and NLF2 belong to a novel gene family encoding nuclear factors with a role in regulating genes which control cellular architecture. This might increase vascular permeability in acute inflammation. PMID- 15527970 TI - Expression of two protein isoforms of PAX7 is controlled by competing cleavage polyadenylation and splicing. AB - The PAX7 gene encodes an evolutionary conserved transcription factor that is involved in the determination of the myogenic cell lineage during the development of vertebrates. In the postnatal period, the function of PAX7 is ultimately required for the specification of muscle satellite cells. The fact that PAX7 is expressed in fast proliferating embryonal myoblasts and in quiescent satellite cells of adults raised the question whether different PAX7 protein isoforms may have distinct roles in these myogenic precursors. Previously, we identified a human PAX7 mRNA encoding a C-terminus which did not show any sequence similarity to the PAX7 proteins of other organisms. So far, there was no further information available concerning the biological nature and significance of this form of PAX7. Here, we show that expression of PAX7 can be regulated by differential transcriptional termination either in exon 9 or in exon 8. Thereby, differential mRNA cleavage-polyadenylation and splicing of PAX7 may result in production of two alternative protein forms that contain or exclude the evolutionary conserved carboxy-terminal domain, respectively. The existence of both protein isoforms in vivo was confirmed by Western blot analysis. These data imply that the alternative C-termini of PAX7 may convey different functions to the corresponding protein isoforms. PMID- 15527971 TI - Functional characterisation and genomic analysis of an epithelial calcium channel (ECaC) from pufferfish, Fugu rubripes. AB - An orthologue to the mammalian epithelial calcium channels, ECaC1 (TRPV5) and ECaC2 (TRPV6), was cloned from gill of pufferfish (Fugu rubripes) and characterised, demonstrating that this gene predates the evolution of land-living vertebrates. The F. rubripes ECaC (FrECaC) protein displays all structural features typical for mammalian ECaCs including three ankyrin repeats, six transmembrane domains, and a putative pore region between TM V and TM VI. Functional expression of FrECaC in Madin-Darby canine kidney (MDCK) cells confirmed that the channel mediates Ca(2+) influx. FrECaC was also permeable to Zn(2+) and, to a small extent, to the Fe(2+) ion. Thus, in addition to a role in Ca(2+) uptake FrECaC might serve as a pathway for zinc and iron acquisition. FrECaC mRNA was highly abundant in the gill, but sparsely present in the intestine. Calcium absorption via FrECaC in pufferfish may be subject to the regulation of 1.25(OH)(2)D(3), estrogen and progesterone as consensus cis regulatory elements for the respective steroid hormone receptors were found in the upstream regulatory region of the FrECaC gene. FrECaC gene organisation is very conserved when compared with mammalian ECaCs. Only one ECaC gene seems to exist in the F. rubripes genome, and the corresponding protein clusters together with ECaC2 from mammals upon phylogenetic analysis. Thus, the two mammalian ECaC genes may originate from a single ancestral ECaC2 gene in vertebrates appearing early in evolution. PMID- 15527972 TI - WD dipeptide motifs and LXXLL motif of chicken HIRA are essential for interactions with the p48 subunit of chromatin assembly factor-1 and histone deacetylase-2 in vitro and in vivo. AB - We cloned cDNA encoding chicken HIRA, a homolog of Saccharomyces cerevisiae transcriptional corepressors Hir1p and Hir2p, possessing seven WD dipeptide motifs and a LXXLL motif in its N-terminal and C-terminal regions, respectively. It binds to CAF-1p48, HDAC-1 and 2, but not to CAF-1p60, p46 polypeptide and HDAC 3. The immunoprecipitation experiment involving truncated and missense mutants of HIRA and CAF-1p48 revealed not only that even one of seven WD dipeptide motifs in the N-terminal half of HIRA are necessary for the interaction with CAF-1p48, but also that those of CAF-1p48 are necessary for the interaction with HIRA. These findings indicate that the proper propeller structures of both HIRA and CAF-1p48 are necessary for their in vitro interaction. The immunoprecipitation experiment involving truncated and missense mutants of HIRA and HDAC-2 revealed that the LXXLL motif in the C-terminal half of HIRA and a C-terminal region of HDAC-2 are necessary for their in vitro interaction. Moreover, the WD dipeptide motifs and LXXLL motif of HIRA are essential for the interaction with CAF-1p48 and HDAC-2 in vivo. Taken together, these results indicate that HIRA should participate differentially in a number of DNA-utilizing processes through interactions of its distinct regions with these proteins. PMID- 15527973 TI - Transcriptional regulation of the MAIL gene in LPS-stimulated RAW264 mouse macrophages. AB - IkappaB inhibits nuclear factor kappa B (NF-kappaB), which is known to regulate the expression of various genes, including genes involved in inflammation. Recently, a novel IkappaB family protein, 'molecule possessing ankyrin repeats induced by lipopolysaccharide' (MAIL), was identified. MAIL is a nuclear-acting, inducible protein, unlike typical IkappaB proteins. However, the mechanism of its induction by lipopolysaccharide (LPS) is unclear. Using the LPS-reactive region located upstream from the MAIL gene, we investigated the mechanism of MAIL induction. MAIL expression was strongly regulated by NF-kappaB and partly regulated by CREB. Furthermore, deletion, point mutation and binding analyses revealed that the NF-kappaB binding site located at -229 to -220 bp is an essential target of MAIL expression. Overexpression of MAIL protein suppressed the LPS-induced promoter activity of the MAIL gene. These data indicate that MAIL expression is strongly upregulated by NF-kappaB, and it is controlled, at least in part, by an autoregulation mechanism. PMID- 15527974 TI - Characterization of promoter elements involved in the down-regulation of topoisomerase IIalpha expression in a drug-resistant cell line. AB - Reduced expression of topoisomerase II is one of the mechanisms observed in cell lines and clinical samples that are resistant to topoisomerase II-targeting agents. The Chinese hamster lung cell line DC-3F/9-OH-E made resistant to 9-OH ellipticine and cross-resistant to other topoisomerase II inhibitors has previously been shown to express lower level of topoisomerase IIalpha isoform, than the parental DC-3F cell line. We have shown here that topoisomerase IIalpha promoter activity is lower in the resistant cell line. The promoter sequence responsible for the differential expression of Chinese hamster topoisomerase IIalpha gene was localized in a small promoter region, which harbors three inverted CAAT elements (ICEs) that bind transcription factor NF-Y, two GC boxes that bind Sp1 and a TATA-like element that binds unknown factors. Immunoblot analysis of cell lysates showed that the resistant line expressed reduced levels of NF-Y subunits and attenuated level of p53. Although p53 has been reported being involved in the regulation of topoisomerase II expression, it is not responsible for the reduced topoisomerase IIalpha expression in the drug resistant line. Mutational analysis of individual elements suggested that the resistant cell line has relaxed responses to ICE mutations, and the TATA-like element plays a predominant role in the regulation of topoisomerase IIalpha. Furthermore, gel mobility shift assays showed that the resistant line has a differential binding to the novel TATA-like element, which may be responsible for the down-regulation of topoisomerase IIalpha gene. PMID- 15527975 TI - Woodchuck interleukin-6 gene: structure, characterization, and biologic activity. AB - Woodchuck is an important animal model for studying human hepatitis B virus (HBV) infection. Within the cytokine network, interleukin-6 (IL-6) plays an important role in immune responses that may lead to viral clearance. To further understand woodchuck IL-6 biology, we cloned and characterized the IL-6 gene from white blood cells. The complete woodchuck IL-6 gene is about 7 kb and consists of five exons and four introns. The IL-6 gene organization of the woodchuck is similar to those of the human, rat, and mouse. Also several elements are highly conserved in the 300 bp promoter region of the IL-6 gene, including a nuclear factor kappa B (NF-kappaB) binding site. The woodchuck IL-6 gene encodes a polypeptide of 207 amino acids in a precursor form and 189 amino acids in the mature form. The expressed protein was 23 kDa according to SDS-PAGE. To demonstrate biologic activity, we expressed woodchuck IL-6 and showed that the purified recombinant protein induced terminal differentiation, as reflected by upregulation of Fcgamma receptor expression, and substantially inhibited proliferation of M1 cells, a murine myeloid leukemia cell line. The inhibitory effect of woodchuck IL-6 on M1 cells was blocked by an anti-gp130 monoclonal antibody, suggesting that woodchuck IL-6 activity is specifically mediated by signaling through the IL-6 receptor complex. Cloning of the woodchuck IL-6 gene and demonstrating biologic activity of the gene product will facilitate studies of human hepatitis B virus using the woodchuck model. PMID- 15527976 TI - Genomic organization of the mouse Msh4 gene producing bicistronic, chimeric and antisense mRNA. AB - By degenerate PCR and screening of mouse testis cDNA library, we have identified seven cDNAs from the meiotic recombination gene Msh4. Variant alpha and probably beta are likely involved in meiotic DNA recombination. Other variants have distinctive structures; variant epsilon, theta; and iota form a bicistronic operon, while variant delta contains antisense RNA for the endoplasmic reticulum (ER) chaperon Hspa5 gene and small open reading frame (ORF) identical to epsilon ORF2. Analysis of the exon-intron structures revealed unusual genomic organizations: the first three exons of delta and the first exon of epsilon are respectively mapped to the Hspa5 locus on chromosome 2 and the Pcbp3 locus on chromosome 10; the remaining exons of both variants are mapped to the Msh4 locus on chromosome 3. The first exon of variant beta is located on chromosome 16, while the others are located on chromosome 3. Synthesis of these mRNAs is assumed to require interchromosomal trans-splicing alone (beta and epsilon) or in combination with converse-splicing (delta). Most Msh4 variant mRNAs are mainly expressed in testis, but a small amount of each variant except for epsilon is also expressed in brain, heart, thymus, ovary and embryonic head. Enhanced green fluorescent protein (EGFP) fusion experiments showed that all the ORFs are translated, and most of those proteins are localized to a particular subcellular compartment. It also appeared that expression of variant delta induces cell death. This study suggests that the dynamic interchromosomal (intergenic) trans splicing generates functional diversity of the mouse Msh4 gene. PMID- 15527977 TI - Identification of an operon involved in tyrosinase activity and melanin synthesis in Marinomonas mediterranea. AB - The genomic region of Marinomonas mediterranea containing the genes required for tyrosinase activity and melanin synthesis has been cloned by marker rescue using the transposon-generated, amelanogenic strain T105. Five ORFs, two incomplete and three complete, have been sequenced in the genomic region where the transposon was inserted. RT-PCR analysis indicates that ORF 3, coding for tyrosinase, and ORF4, coding for a protein of 250 amino acids, are in the same transcriptional unit, constituting an operon whose promoter region has been determined by 5' RACE. This operon has been sequenced in the wild-type and several mutant strains, indicating that both ORFs are required for expression of tyrosinase activity and melanin synthesis. The nitrosoguanidine generated, amelanogenic mutant ng56, shows a nonsense mutation in ORF3 coding for the tyrosinase. On the other hand, in the strain T105 the transposon is inserted in ORF4. The product of this gene is related to copper metabolism, since the addition of this metal ion to cell extracts or culture media partially restores melanin synthesis and tyrosinase activity in the strain T105. However, it does not show significant sequence similarity to previously characterized metallochaperones and hence may be an example of a new kind of those proteins. The operon has been denoted as ppoB, taking into consideration that ppoA denotes the M. mediterranea gene coding for the previously cloned polyphenol oxidase with laccase activity. This is the first demonstration of the tyrosinase gene forming part of an operon in a Gram-negative bacterium. PMID- 15527978 TI - Identification and phenotypic characterization of the cell-division protein CdpA. AB - Analysis of the automated computer annotation of the early draft phase genome of Lactobacillus acidophilus NCFM revealed the previously discovered S-layer gene slpA and an additional partial ORF with weak similarities to S-layer proteins. The entire gene was sequenced to reveal a 1799-bp gene coding for 599 amino acids with a calculated molecular mass of 64.8 kDa. No transcription or translation signals could be determined in close proximity to the 5'-region. However, a strong putative terminator with a free energy of -16.84 kcal/mol was identified directly downstream of the gene. A PSI-Blast analysis showed similarities to members of S-layer proteins, cell-wall associated proteinases and hexosyl transferases. Calculation of an unrooted phylogenetic tree with other examples of S-layer proteins and proteinases placed the deduced protein separately from both groups. A derivative of L. acidophilus NCFM was constructed by targeted integration into the gene. SDS-PAGE analysis of non-covalently linked proteins of the cell wall of the mutant, compared to the wild type, revealed the loss of a cell-surface protein. Phenotypic analyses of the mutant revealed significant changes in cell morphology, altered responses to various environmental stresses, and lowered cell adhesion. Based on the in silico and functional analyses, we ascertained that this protein plays a role in cell-wall processing during the growth and cell-cell separation and designated the gene as cell-division protein, cdpA. PMID- 15527979 TI - Characterization of the two arginine decarboxylase (polyamine biosynthesis) paralogues of the endemic subantarctic cruciferous species Pringlea antiscorbutica and analysis of their differential expression during development and response to environmental stress. AB - Arginine decarboxylase (ADC) is a key enzyme involved in the synthesis of polyamines, which have been implicated in developmental processes and stress responses in higher plants. An ancestral ADC gene appears to have been duplicated at the origin of the Brassicaceae family, thus yielding two paralogues in the derived taxa. ADC gene structure was investigated in Pringlea antiscorbutica R. Br., a geographically isolated Brassicaceae species that is endemic from the subantarctic region. P. antiscorbutica exhibits several biochemical and physiological adaptations related to this cold and harsh environment, including high levels of polyamines, which is unusual in higher plants, and especially high levels of agmatine, the product of the ADC-catalysed reaction. Various ADC clones were obtained from P. antiscorbutica. Sequence and phylogenetic analysis showed that all these clones fitted with the presence of two paralogues, PaADC1 and PaADC2, in P. antiscorbutica. Expression of these ADC paralogues was analyzed in P. antiscorbutica during vegetative development and response to stress. Whereas PaADC2 was expressed at both seedling and mature stages, PaADC1 transcripts were hardly detected during early development and were significantly expressed in mature plants. Moreover, PaADC2, but not PaADC1, expression was up-regulated in response to chilling and salt stress at seedling stage. Analysis of 5' regulatory regions of these ADC genes revealed several differences in putative cis regulatory elements, which could be associated with specific expression patterns. These results were compared to ADC paralogue expression in Arabidopsis thaliana and are discussed in the evolutionary context of genetic diversity resulting from gene duplication. PMID- 15527980 TI - Construction and diversification of yeast cell surface displayed libraries by yeast mating: application to the affinity maturation of Fab antibody fragments. AB - Yeast display is a powerful technology for the affinity maturation of human antibody fragments. However, the technology thus far has been limited by the size of antibody libraries that can be generated, as using current transformation protocols libraries of only between 10(6) and 10(7) are typically possible. We have recently shown that Fab antibodies can be displayed on the cell surface of Saccharomyces cerevisiae [van den Beucken, T., Pieters, H., Steukers, M., van der Vaart, M., Ladner, R.C., Hoogenboom, H.R., Hufton, S.E., 2003. Affinity maturation of Fab antibody fragments by fluorescent-activated cell sorting of yeast-displayed libraries. FEBS Lett. 546, 288-294]. This discovery and the knowledge that Fab antibodies are heterodimeric suggest that independent repertoires of heavy chain (HC) and light chain (LC) can be constructed in haploid yeast strains of opposite mating type. These separate repertoires can then be combined by highly efficient yeast mating. Using this approach, we have rapidly generated a naive human Fab yeast display library of over 10(9) clones. In addition, utilizing error-prone polymerase chain reaction, we have diversified Fab sequences and generated combinatorial and hierarchical chain shuffled libraries with complexities of up to 5 x 10(9) clones. These libraries have been selected for higher affinity using a repeating process of mating-driven chain shuffling and flow cytometric sorting. PMID- 15527981 TI - RZF, a zinc-finger protein in the photoreceptors of human retina. AB - Photoreceptors are organized at the outer aspect of retina and host the process of phototransduction, central to the visual system. We have isolated a novel human gene, RZF, which is predominantly expressed in the photoreceptors of human retina. RZF encodes a 40-kDa protein that has three widely spaced C(2)H(2)-type zinc finger motifs. There are three potential nuclear localisation signals and clusters of charged amino acids in the protein. Expression analysis revealed that orthologues of the RZF gene are also expressed in photoreceptors of mouse and bovine retina. The RZF-GFP fusion protein localises to nucleoli and cytoplasm when expressed in HEK-293 cells. Mobility shift assay suggests that RZF may not be a nucleic acid binding protein, unlike most other zinc-finger proteins. Taken together, these observations suggest that RZF is a shuttling regulatory protein expressed in photoreceptors of the human retina that may be involved in mRNA or protein regulation of photoreceptor-specific genes and therefore have role in retinal disease mechanisms. PMID- 15527982 TI - TBP-associated factors in Arabidopsis. AB - Initiation of transcription mediated by RNA polymerase II requires a number of transcription factors among which TFIID is the major core promoter recognition factor. TFIID is composed of highly conserved factors which include the TATA binding protein (TBP) and about 14 TBP-associated factors (TAFs). Since TAFs play important roles in transcription they have been extensively studied in organisms like yeast, Drosophila and human. Surprisingly, TAFs have been poorly characterized in plants. With the completion of the Arabidopsis genome sequence, it is possible to search for TAFs, since many of them have conserved amino acid sequences. Mining the genome of Arabidopsis for TAFs resulted in the identification of 18 putative Arabidopsis TAFs (AtTAFs). We have analyzed their protein structure and their genomic localisation. Expression profiling by RT-PCR showed that these TAFs are expressed in all parts of the plant which is in agreement with their general role in transcription. These analyses in combination with their evolutionary conservation with TAFs of other organisms are discussed. PMID- 15527983 TI - A thrombin inhibitor from the ixodid tick, Amblyomma hebraeum. AB - A novel thrombin inhibitor named Amblin was identified from the haemolymph of the ixodid (hard) tick, Amblyomma hebraeum, and the coding cDNA was isolated from a tick cDNA library. This cDNA codes for a preprotein of 166 amino acids, including a predicted signal peptide composed of 15 amino acids N-terminal to the mature Amblin. The 151-amino-acid mature Amblin contains 14 cysteines and two Kunitz like domains. It displays high sequence similarity with a tissue factor pathway inhibitor (TFPI), Ixolaris, from the ixodid tick, Ixodes scapularis, which has 10 cysteines, and a thrombin inhibitor, Boophilin, from the ixodid tick, Boophilus microplus, which has 12 cysteines. Recombinant Amblin specifically inhibited thrombin as efficiently as native Amblin did. This is the first report of a thrombin inhibitor from tick haemolymph. PMID- 15527984 TI - Src proteins/src genes: from sponges to mammals. AB - The genome of marine sponge Suberites domuncula, a member of the most ancient and most simple metazoan phylum Porifera, encodes at least five genes for Src-type proteins, more than, i.e., Caenorhabditis elegans or Drosophila melanogaster (two in each). Three proteins, SRC1SD, SRC2SD and SRC3SD, were fully characterized. The overall homology (identity+similarity) among the three S. domuncula Srcs (68 71%) is much lower than the sequence conservation between orthologous Src proteins from freshwater sponges (82-85%). It is therefore very likely that several src genes/proteins were already present in the genome of Urmetazoa, the hypothetical metazoan ancestor. We have identified in the S. domuncula expressed sequence tags (ESTs) database further Src homology 2 (SH2) and 3 (SH3) domains that are unrelated to protein tyrosine kinases (PTKs). Src-related SH2 and SH3 domains from different species are much more conserved than SH2 and SH3 domains from different proteins in the same organism (S. domuncula), supporting the view that the common, ancestral src gene was already a multidomain protein composed of SH3, SH2 and tyrosine kinase (TK) domains. Two S. domuncula src genes were fully sequenced: src1SD gene has six and src2SD gene only one intron in front of SH2 domain, located at the same position in both genes. All vertebrate src genes, from fish to human, originated from the same ancestral gene, because they all have 10 introns at conserved positions. However, src genes in invertebrates have fewer introns that are located at different positions. Only the intron in front of the SH2 domain is present at the absolutely conserved position (and phase) in all known src genes, indicating that at least this intron was already present in the ancestral gene, common to all Metazoa. Our results also suggest that TK domain in this ancestral src was encoded on a single exon. PMID- 15527985 TI - Different age distribution patterns of human, nematode, and Arabidopsis duplicate genes. AB - We studied the age distribution of duplicate genes in each of four eukaryotic genomes: human, Arabidopsis thaliana, Caenorhabditis elegans, and Drosophila melanogaster. The four distributions differ greatly from each other, contrary to the previous proposal of a universal L-shaped distribution in all eukaryotic genomes studied. Indeed, only the distribution in humans is L-shaped. The distribution in Arabidopsis is consistent with the hypothesis of an ancient genome duplication with no recent burst of duplication events, while the distribution in C. elegans is nearly uniform. We also applied a nonparametric method to the human distribution to show that the rate of loss of duplicate genes decreases over time, contrary to the proposal of an exponential decay. One possible explanation of the decreasing rate of loss of duplicate genes over time could be rapid functional divergence between duplicate genes, providing an advantage for the retention of both duplicates. PMID- 15527986 TI - AcFKH1, a novel member of the forkhead family, associates with the RFX transcription factor CPCR1 in the cephalosporin C-producing fungus Acremonium chrysogenum. AB - In the filamentous fungus Acremonium chrysogenum, a complex regulatory network of transcription factors controls the expression of at least seven cephalosporin C biosynthesis genes. The RFX transcription factor CPCR1 binds to regulatory sequences in the promoter region of cephalosporin C biosynthesis genes, and is involved in the transcriptional regulation of the pcbC gene which encodes isopenicillin N synthase. In this study, we used CPCR1 in a yeast two-hybrid screen to identify potential protein interaction partners. A cDNA was identified, encoding the C-terminal part (pos. 438-665) of the novel forkhead protein, AcFKH1. The full-length AcFKH1 amino acid sequence is 665 residues and shares between 31% and 60% identity with forkhead protein sequences in the genomes of Aspergillus nidulans, Fusarium graminearum, and Neurospora crassa. AcFKH1 is characterized by two conserved domains, the N-terminal forkhead-associated domain (FHA), which might be involved in phospho-protein interactions, and the C terminal DNA-binding domain (FKH) of the winged helix/forkhead type. The two hybrid system was also used to map the protein domains required for the interaction of transcription factors CPCR1 and AcFKH1. The observed interaction between CPCR1 and the C-terminus of AcFKH1 in the yeast system was verified in vitro in a GST pulldown assay. Using gel retardation analysis, the DNA-binding properties of the fungal forkhead protein AcFKH1 were investigated. AcFKH1 recognizes two forkhead consensus binding sites within the 1.2 kb promoter region of the divergently oriented cephalosporin biosynthesis gene pair pcbAB-pcbC from A. chrysogenum. Additionally, AcFKH1 is able to bind with high affinity to the SWI5-binding site of the yeast FKH2 protein. PMID- 15527987 TI - Human endogenous retroviral elements belonging to the HERV-S family from human tissues, cancer cells, and primates: expression, structure, phylogeny and evolution. AB - A new human endogenous retrovirus family (HERV-S) was recently identified from the human chromosome X. The HERV-S was 6.7 kb in length with typical retroviral structure (LTR-gag-pol-env-LTR). We investigated pol fragments of HERV-S family in various human tissues and cancer cells. The pol gene was expressed in only brain and thymus among human tissues (brain, prostate, testis, heart, kidney, liver, lung, placenta, skeletal muscle, spleen, thymus, uterus), whereas the pol gene was detected in various cancer cell lines (RT4, PFSK-1, BT-474, HCT-116, Jurkat, HepG2, MCF7, OVCAR-3, MIA-PaCa-2, PC3, LOX-IMVI, AZ521, 2F7, and C-33A) except for TE-1, UO-31, A549, and U-937 by RT-PCR analysis. Expression and sequencing data of the 33 clones imply that the pol gene of HERV-S family is more active in cancer cells than in human tissues, which may have transcriptional potential role related to various human cancers. Phylogenetic analysis of HERV-S pol family distinctively divided into three groups (group I, II, and III) through evolutionary divergence in primate evolution. Evolutionary divergence of the HERV S family by PCR amplification and sequence analysis indicated that they were integrated into the primate genomes approximately 43 Myr ago and have been evolved rate of 0.3% nucleotide differences per Myr during primate evolution. PMID- 15527988 TI - Promoter and piggyBac activities within embryos of the potato tuber moth, Phthorimaea operculella, Zeller (Lepidoptera: Gelechiidae). AB - Potato production in tropical and subtropical countries suffers from damage caused by the potato tuber moth (PTM), Phthorimaea operculella. The aim of this research was the development of the components required for a germline transformation system for the PTM. We tested three components that are critical to genetic transformation systems for insects: promoter activity, marker gene expression, and transposable element function. We compared the transcriptional activities of five different promoters, hsp70, hsp82, actin5C, polyubiquitin and immediate early 1 gene (ie1), within PTM embryos. The ie1 promoter, flanked by the hr5 enhancer element, showed a very high level of transcriptional activity compared to the other promoters. The fluorescence activity of EGFP was also determined and transient expression of EGFP was detected in 57% of injected embryos. The transpositional activity of the piggyBac transposable element was tested in an interplasmid transposition assay. The piggyBac element was shown to be mobile within the embryonic soma of the PTM with a transposition frequency of 4.2 x 10(-5) transpositions/donor plasmid. Incorporating a transactivator plasmid expressing the immediate early protein (IE1) from the Bombyx mori nuclear polyhedrosis virus enhanced the efficiency of piggyBac mobility. PMID- 15527989 TI - Desaturases from the spotted fireworm moth (Choristoneura parallela) shed light on the evolutionary origins of novel moth sex pheromone desaturases. AB - Six acyl-CoA desaturase-encoding cDNAs from mRNA isolated from the spotted fireworm moth, Choristoneura parallela (Lepidoptera: Tortricidae) were characterized and assayed for functionality. The expression levels of these cDNAs were determined in the pheromone gland and fat body by real-time PCR and the resulting patterns are in line with results from published studies on other moth sex pheromone desaturases. The cDNAs were found to correspond to six genes. Using both biochemical and phylogenetic analyses, four of these were found to belong to previously characterized desaturase functional groups [the Delta 10,11, the Delta 9 (16>18) and the Delta 9 (18>16) groups]. A desaturase highly expressed in the pheromone gland was a novel E11 desaturase that was specific to 14-carbon precursor acids. The fifth gene [CpaZ9(14-26)] was found to display a novel Z9 activity indicating that it belongs to a new Delta 9 functional group, whereas the sixth gene was determined to be nonfunctional with respect to desaturase activity. In accordance with previous studies, we find that desaturases of the Delta 10,11 and Delta 14 groups, which are the fastest evolving desaturases and possess the novel pheromone biosynthetic function, are expressed primarily in the pheromone gland whereas all other desaturases, which do not possess the novel reproductive function, evolve more slowly and display the ancestral metabolic function and pattern of gene expression. PMID- 15527990 TI - Genomic dynamics of a low-copy-number satellite DNA family in Leptestheria dahalacensis (Crustacea, Branchiopoda, Conchostraca). AB - The LEP150 satellite DNA (satDNA) family found in Leptestheria dahalacensis (Ruppel, 1837) (Conchostraca) is a low-copy-number satellite with a canonical monomer of 150 bp. Nucleotide variation analyses suggest a 14-bp palindromic region as a possible protein binding site with constraints acting on the whole sequence but a 25-bp variable box. Besides the head-to-tail arrangement of 150 bp monomers, multimers analyses evidenced incomplete monomers, one duplication event, and three inversions. Both observed rearrangements and the higher values of sequence variability scored suggest that rearranged monomers reside in regions with a lower degree of homogenisation efficiency. Sixty-seven percent of the breakpoints occurs at kinkable dinucleotides, thus supporting their role in rearrangements as documented in alphoid satDNA recombination events. Monomers of different lengths may result from crossing over between repeats misaligned through the direct and inverted subrepeats of LEP150 monomers. ANOVA results indicate that the same range of sequence diversity is experienced at the individual and population ranks; therefore, the evolution of the L. dahalacensis satDNA is concerted. PMID- 15527991 TI - An analysis of sponge genomes. AB - The genome of sponges has only been investigated so far by Bartmann-Lindholm et al. [Progr. Colloid. Polym. Sci. 107 (1997) 122-126] who reported a multimodal CsCl profile which could be resolved into five peaks for Geodia cydonium. This problem was reinvestigated here on both G. cydonium and Suberites domuncula. It was shown that DNAs from both sponges are characterized by unimodal CsCl profiles, additional peaks being due to contaminating prokaryotic and eukaryotic microorganisms. PMID- 15527992 TI - Magnetic resonance imaging of the knee with ultrashort TE pulse sequences. AB - BACKGROUND: We wished to assess the feasibility of imaging the knee with ultrashort TE (UTE) pulse sequences. SUBJECTS AND METHODS: Five volunteers and 16 patients were studied with UTE (TE=0.08 ms) sequences including later echoes. Conventional fat-suppressed images and difference images were also produced by subtracting a later echo from the first. Gadodiamide enhancement was used. RESULTS: High signal was obtained in tendons, ligaments, menisci and periosteum. Normal contrast enhancement was seen in these structures. Deep and superficial layers were seen in the articular cartilage. Cartilage defects were identified. The red zone could be differentiated from the white zone of the meniscus. Meniscal tears and degeneration were observed with low signal on subtraction images. Enhancement was seen within the anterior and posterior cruciate ligaments and associated scar tissue. CONCLUSION: Ultrashort TE imaging provides new options to visualize anatomy, manipulate conspicuity, observe contrast enhancement and demonstrate disease of the knee. PMID- 15527993 TI - The effect of sensorimotor activation on functional connectivity mapping with MRI. AB - The correlations in the fluctuations in the blood oxygenation level-dependent (BOLD) MRI signal between anatomically distinct regions of the cortex that are known components of functional systems have been previously studied as possible indicators of functional connectivity. The objective of this study was to examine the effect of sensorimotor brain activity, as assessed by task-based functional magnetic resonance imaging (fMRI), on functional connectivity indices in the same region. Regions of activation for sequential finger motion were determined using a task-based, block-design fMRI study. Functional connectivity measurements based on interregional correlations were acquired at rest and during continuous, sequential finger motion. Connectivity indices were determined using normalized mean correlations within and between three regions of interest activated for the finger motion task. Connectivity indices were also determined for a control region that was not activated for the task. Continuous motor tasks performed during BOLD measurements did not significantly affect the functional connectivity as compared to the connectivity at rest within or between regions known to be activated by the task. However, there appeared to be a trend suggesting a slight reduction in connectivity indices during the motor task. The connectivity within and between those areas not activated for the task remained unchanged between conditions. These results suggest that in the motor system investigated, the recruitment of neurons to perform a specific task may moderately reduce the degree of hemodynamic coupling within and between regions. PMID- 15527994 TI - Imaging of a mixture of hyperpolarized 3He and 129Xe. AB - With the use of hyperpolarized gases, a great number of experiments have been carried out in order to improve the diagnostics of the lung, both from a structural and a functional point of view. 3He is best suited for structural studies, whereas 129Xe gives more detailed information about the functionality of the lung because it enters the bloodstream. In this work, we propose the use of a gas mixture to perform consecutive analysis of lung structure and functionality upon the delivery of a single bolus of gas. We show images of a helium-xenon gas mixture in the presence of a small amount of liquid toluene in order to demonstrate how both nuclei can be detected independently, extracting the spectroscopic information provided by the 129Xe spectra and obtaining an image with high sensitivity for 3He. A second experiment performed on a dissected mouse lung was used to demonstrate how the mixture of gases can enhance sensitivity in the larger airways of the lung. PMID- 15527995 TI - MR enteroclysis using iron oxide particles (ferristene) as an endoluminal contrast agent: an open phase III trial. AB - To evaluate efficacy and safety of a superparamagnetic iron oxide contrast agent (ferristene) as an endoluminal contrast medium for magnetic resonance (MR) enteroclysis in a phase III trial. Twenty-three patients with history of known or suspected small bowel Crohn's disease underwent MR imaging of the abdomen at 0.5 T unit. The imaging protocol included two phases: the first one without administration of any contrast agent and the second one, where the small bowel was filled by enteroclysis with 800 ml of the luminal iron oxide contrast medium and Gd-DTPA (0.1 mmol/Kg) was administered intravenously. Axial Spin-Echo (SE) T1 weighted (T1w), proton-density and T2w images, sagittal and coronal SE T1w and Short TI Inversion Recovery (STIR) sequences were subsequently obtained. Three investigators blindly evaluated images to determine small bowel distribution of ferristene, presence of artifacts, delineation of bowel lesion/wall and the diagnostic value of ferristene combined with gadolinium. Pre- and postcontrast signal intensity measurements of bowel lesion/wall, bowel lumen and background noise were also calculated. Three patients withdrew before the procedure, therefore 20 patients were effectively included in the study. No significant difference between the three investigators' evaluations of the improvement of the diagnostic information was found (percentage of improvement of 90% with 95% confidence limits of 68% and 99%). A statistically significant difference between the first and third investigators was found for grading of quality of delineation of bowel lesion/wall. Signal intensity measures showed a significant increase of the bowel lesion/wall and background noise/lesion for the SE T1w images. No serious adverse event was reported in our series. MR enteroclysis using ferristene as an endoluminal contrast agent appears to be a safe and efficient procedure for the study of the small bowel. PMID- 15527996 TI - MRI of hip prostheses using single-point methods: in vitro studies towards the artifact-free imaging of individuals with metal implants. AB - Use of magnetic resonance imaging (MRI) in individuals with orthopedic implants is limited because of the large distortions caused by metallic components. As a possible solution for this problem, we suggest the use of single-point imaging (SPI) methods, which are immune to the susceptibility artifacts observed with conventional MRI methods. A further advantage of SPI, based on the fact that signal encoding is achieved in ultra-short times (as short as tens of microseconds), is that they enable the direct visualization of the polymeric elements of the implants, allowing the detection of possible implant failures. We present in vitro SPI images of polymeric sockets of two hip prostheses together with artifact-free images of gelatin phantoms containing their respective metallic stems. These data underscore the great potential of the SPI technique for obtaining artifact-free images of individuals with large metal implants. PMID- 15527997 TI - Computer-aided quantification of focal cartilage lesions of osteoarthritic knee using MRI. AB - Noninvasive assessment of articular cartilage using magnetic resonance imaging (MRI) has gained popularity in the diagnosis of osteoarthritis (OA), a condition that affects 20 million Americans. Focal cartilage lesions, a defect found in roughly 19% of the OA population, currently can only be evaluated with confidence using minimally invasive arthroscopy. This article presents a computer-aided procedure using MRI to quantify focal cartilage lesions and aims to support clinical practices of diagnosis and monitoring of lesion progress. Upon a local minima search for identifying focal lesions, the proposed gradient peak method outlines lesion boundaries and then generates morphological properties, such as lesion volume and lesion area. The procedure was evaluated using simulated and in vivo data. First, a simulated lesion was created and analyzed, and the results were compared with the exact solutions. Second, an in vivo evaluation was carried out on seven human knees in which nine focal lesions were identified and quantified. Three of the subjects had follow-up analyses, at either 1 or 2 years. Finally, in an attempt to characterize local biochemical changes underlying focal lesions, MR-derived T2 values of defective cartilage within the lesion boundaries were examined and compared with the values of adjacent cartilage compartments. PMID- 15527998 TI - A study of the relationship between molecular biomarkers of joint degeneration and the magnetic resonance-measured characteristics of cartilage in 16 symptomatic knees. AB - We used quantitative magnetic resonance (MR) imaging to determine if relationships exist between proposed molecular biomarkers for degenerative joint disease (DJD) and structural characteristics of articular cartilage. Subjects were eight male and eight female volunteers diagnosed with osteoarthritis. Magnetic resonance images of the symptomatic knee were taken and blood samples were drawn. Concentrations of serum cartilage oligomeric matrix protein (COMP) and cleaved collagen neoepitope were compared to cartilage volume and cartilage T2, respectively, in four compartments of the tibiofemoral joint. A significant, negative correlation was found between serum COMP and medial tibia volume in the male subject group (rho=-.738, P=.037). A significant, positive correlation (rho=.881, P=.0039) was found between serum COMP and lateral femur volume in the female subject group. In both groups, positive correlations were found between serum C2C and cartilage T2, which were significant in two compartments of the male group (rho=.714, P=.047; rho=.738, P=.037) and similarly strong, but not statistically significant (rho=.750, P=.052), in one compartment of the female group. We identify strong and biologically relevant correlations between two proposed molecular biomarkers for DJD and MR measures of symptomatic knees of a small number of arthritic patients. Our findings support the hypothesis that cartilage molecular biomarkers reflect the molecular processes of cartilage degeneration and loss. PMID- 15527999 TI - Anterior cruciate ligament graft impingement against the posterior cruciate ligament: diagnosis using MRI plus three-dimensional reconstruction software. AB - OBJECTIVE: The purpose of this study was to evaluate anterior cruciate ligament (ACL) impingement against the posterior cruciate ligament (PCL) with the knee in an extended position, which arthroscopy cannot detect. MATERIALS AND METHODS: Ten normal knees and 30 ACL-reconstructed knees were assessed using MR imaging. The three-dimensional reconstruction of the ACL, PCL, femur and tibia were carried out using commercially available three-dimensional reconstruction software. Anterior cruciate ligament impingement against the PCL was graded into three categories: Grade 1, some space between the ligaments; Grade 2, no space between the ligaments, and the reconstructed ACL ran straight; and Grade 3, the reconstructed ACL did not run straight. The angle of the reconstructed ACL against the tibial plateau was also measured. RESULTS: All normal knees were classified as Grade 1. The 30 reconstructed knees were classified as follows: Grade 1, 12 cases; Grade 2, 7 cases; and Grade 3, 11 cases. The mean angle of the Grade 3 reconstructed ACL knees was significantly more vertical against the tibia as compared with the Grade 1 knees (P<.05). The postoperative KT-2000 side-to side difference of the Grade 3 knees (2.8+/-4.5 mm) was larger than that of the Grade 1 knees (0.2+/-1.7 mm) and Grade 2 knees (-0.6+/-2.2 mm), but no statistically significant difference could be detected between the three groups in the postoperative KT-2000 data. CONCLUSION: This method is useful to evaluate ACL impingement against PCL, which cannot be detected by conventional arthroscopy during the operation. The surgeon should pay careful attention to the coronal angle of the reconstructed ACL. PMID- 15528000 TI - Is 'virtual histology' the next step after the 'virtual autopsy'? Magnetic resonance microscopy in forensic medicine. AB - AIM: The study aimed to validate magnetic resonance microscopy (MRM) studies of forensic tissue specimens (skin samples with electric injury patterns) against the results from routine histology. METHODS AND RESULTS: Computed tomography and magnetic resonance imaging are fast becoming important tools in clinical and forensic pathology. This study is the first forensic application of MRM to the analysis of electric injury patterns in human skin. Three-dimensional high resolution MRM images of fixed skin specimens provided a complete 3D view of the damaged tissues at the site of an electric injury as well as in neighboring tissues, consistent with histologic findings. The image intensity of the dermal layer in T2-weighted MRM images was reduced in the central zone due to carbonization or coagulation necrosis and increased in the intermediate zone because of dermal edema. A subjacent blood vessel with an intravascular occlusion supports the hypothesis that current traveled through the vascular system before arcing to ground. CONCLUSION: High-resolution imaging offers a noninvasive alternative to conventional histology in forensic wound analysis and can be used to perform 3D virtual histology. PMID- 15528002 TI - The HALO system--a light weight portable imaging system. AB - This paper describes the development of a prototype portable magnetic resonance imaging (MRI) device with anticipated application in material science and biomedicine. The device consists of two rings constructed from permanent magnets with the polarization aligned axially. The geometry of the device produces a B0 field with cylindrical symmetry. A region of homogeneous field of 50 mT has been achieved. Images of small phantoms and one biological sample are presented to demonstrate the capabilities of the device. PMID- 15528001 TI - Remyelination assessment by MRI texture analysis in a cuprizone mouse model. AB - The purpose of this study was to perform serial texture analysis of brain MRI of cuprizone-treated mice for the assessment of regional demyelination and remyelination. Cuprizone-fed mice undergo a brain demyelination process. This process was followed over 56 days by MRI in the olfactory bulbs, cerebellum, putamen and brain stem. The texture of T2-weighted images has been analyzed at two levels: (1) with the average intensity as first order parameter and (2) with several higher order parameters for the best differentiation between myelinated (controls) and demyelinated brains. The most pertinent of these parameters, called horizontal gray level nonuniformity (HGLNU), has been selected by stepwise discriminant analysis. The time evolution of the average value of HGLNU not only confirmed the overall demyelination tendency followed by the average intensity, but also more precisely characterized a transitory remyelination on day 41 in the olfactory bulbs and cerebellum, in agreement with already published immunohistochemical destructive studies. PMID- 15528003 TI - Autoimmune hemolytic anemia: magnetic resonance findings. AB - Idiopathic autoimmune hemolytic anemia (AIHA) is a rare hematological disease typically characterized by extracellular hemolysis. An unusual case of AIHA with interesting magnetic resonance imaging (MRI) appearances is presented. Possible explanations for the MRI findings are discussed. PMID- 15528004 TI - Adrenal adenoma with organizing hematoma: diagnostic dilemma at MRI. AB - We report a case of adrenal adenoma with organizing hematoma mimicking hemangioma on magnetic resonance imaging (MRI). The lesion demonstrated heterogeneous hyperintensity on heavily T2-weighted images. On dynamic contrasted-enhanced MRI, the lesion demonstrated early, patchy peripheral enhancement with subsequent fill in that persisted. Chemical shift gradient-echo images failed to demonstrate the presence of intracellular lipid. Magnetic resonance imaging failed to characterize the lesion, and an erroneous preoperative diagnosis of adrenal hemangioma was made. Although the MRI findings reflected the organized hematoma with abundant vascular spaces, our case emphasizes the point that the MRI characteristics of intratumoral hemorrhage may overlap with those of adrenal hemangioma and chronic expanding hematoma. PMID- 15528005 TI - Ethanol intake inhibits growth of the epithelium in the intestine of pregnant rats. AB - The effects of ethanol ingestion on jejunal and ileal epithelial cells were studied in pregnant rats by measuring crypt and villus cell population, crypt cell proliferation, and crypt cell cycle time as parameters. Timed-pregnancy female rats were fed a liquid diet containing either ethanol [designated as ethanol-fed group (EFG)] or an isocaloric amount of carbohydrate [designated as pair-fed group (PFG)] from gestational day 2 up to delivery. Daily diet ingestion, body weight, nitrogen balance, and nitrogen digestibility were assessed during the gestational period. Crypt and villus cell population, crypt cell proliferation, and crypt cell cycle time were measured in the maternal small intestine at the time animals were killed, just after delivery. Ethanol consumption resulted in ileal hypoplasia of the crypt and villus, but only the villus showed hypoplasia in the jejunum. In addition, crypt cell proliferation was markedly decreased, whereas crypt cell cycle time was longer, both in the jejunum and ileum of the EFG. Ethanol ingestion had no significant effect on body weight gain, nitrogen balance, and nitrogen digestibility. According to our expectations, the offspring from the EFG had significantly lower body weight. In conclusion, chronic ethanol ingestion during pregnancy inhibited the maternal intestinal epithelium growth, more extensively in the ileum. PMID- 15528006 TI - Ethanol intake is increased by injection of galanin in the paraventricular nucleus and reduced by a galanin antagonist. AB - Ethanol intake stimulates expression of galanin in several hypothalamic sites, including the paraventricular nucleus. Because injection of galanin in the paraventricular nucleus induces eating, we hypothesized that galanin might also affect ethanol intake. Rats were given ad libitum access to 4% ethanol for 4 weeks and assigned to one of two groups according to levels of ethanol consumption: high levels (>1.5 g/ kg/day) or low levels (<1.0 g/kg/day). In Experiment 1, galanin (1.0 nmol) or Ringer's solution was injected unilaterally into the paraventricular nucleus, with food and water absent during the first 4 h. Galanin significantly increased ethanol intake only in rats that drank high levels of ethanol. In Experiment 2, injection of galanin (0.5 and 1.0 nmol) in the paraventricular nucleus dose-dependently increased ethanol intake with food and water available. The higher dose was also effective in eliciting ethanol intake when tested with food and water absent. In Experiment 3, a test of receptor specificity was provided by injecting rats with the galanin antagonist M 40 (0.5 nmol) or Ringer's solution. Injection of M-40 in the paraventricular nucleus significantly decreased ethanol consumption. In Experiment 4, an anatomic control, with galanin injected 2 mm dorsal to the paraventricular nucleus in the same animals, caused no change in ethanol intake. In conclusion, injection of galanin in the paraventricular nucleus, at a dose known to induce feeding, acted by means of a galanin receptor to potentiate intake of 4% ethanol, even with food and water available as alternate sources of calories and fluid, respectively. Because ethanol can increase expression of galanin mRNA in the paraventricular nucleus, this could set the stage for a positive feedback loop between galanin and ethanol intake. PMID- 15528007 TI - EEG alpha variants and alpha power in Hispanic American and white non-Hispanic American young adults with a family history of alcohol dependence. AB - Results from several studies support associations among variants in electroencephalogram (EEG) alpha voltage, alcohol dependence, ethnic heritage, and a family history of alcohol dependence, although no studies have as yet been conducted in individuals of Hispanic ethnicity. Categorization of EEG, by using spectral analyses, into high-, medium-, and low-voltage alpha, as well as absolute EEG power in alpha and beta frequency ranges, was carried out for Hispanic American (n=108) and white non-Hispanic American (n=269) young adult men and women (age range, 18 to 25 years) who had a family history, but no personal history, of alcohol dependence. Eighteen percent (n=70) of the participants were categorized as having a low-voltage alpha EEG, 16% had high-voltage alpha, and 64% had medium-voltage alpha. In comparison with findings for men, women were found to have higher overall power in the low frequency alpha (7.5-9 Hz) and beta (12-20 Hz, 20-50 Hz) frequency ranges. Hispanic Americans had a lower percentage of individuals in the low-voltage alpha group (9%) and more individuals in the medium-voltage alpha group (73%) compared with findings for white non-Hispanic Americans (low-voltage alpha, 19%; medium-voltage alpha, 62%). Results of the current study confirm the presence of ethnic stratification in EEG alpha variants. PMID- 15528008 TI - Effects of concurrent access to a single concentration or multiple concentrations of ethanol on ethanol intake by periadolescent high-alcohol-drinking rats. AB - The objectives of the current study were to assess the effects of access to different concentrations of ethanol and sex of the animal on ethanol consumption of high-alcohol-drinking (HAD-1 and HAD-2) rats during adolescence [postnatal days (PNDs) 30 through 60]. At the beginning of adolescence (PND 30), the rats were given concurrent access to either a single concentration [15% volume/volume (vol./vol.)] or multiple concentrations [10%, 20%, and 30% (vol./vol.)] of ethanol and water. Analyses of ethanol consumption data revealed significant (P < .025) main effects of line, ethanol condition, and week, and a significant line by sex by ethanol condition by week interaction. For the first week, both male and female HAD-1 and HAD-2 rats consumed more ethanol under the multiple ethanol concentration condition than under the single ethanol concentration condition. However, across the second through fourth weeks, this pattern was seen primarily in male and female HAD-1 rats and to a lesser degree in female HAD-2 rats. In general, female rats consumed more fluids than consumed by male rats, and male rats displayed a higher preference for ethanol over water ratio than observed for their female counterparts. In addition, in comparison with HAD-2 rats, HAD-1 rats drank more ethanol and displayed a higher preference for ethanol ratio. Overall, the current study results indicate that, compared with access to a single concentration (which is used in most studies), concurrent access to multiple concentrations of ethanol produced significantly higher ethanol intakes in periadolescent HAD rats, supporting the suggestion that this ethanol drinking condition would have a greater impact on neuronal development. In addition, although the replicate lines were selectively bred by using the same criteria and foundation stock, the higher ethanol intakes of the HAD-1 line, compared with intakes for the HAD-2 line, seen in the current study support the suggestion that there are some differences in their genetic make-up, affecting ethanol intake, which are expressed during periadolescence. PMID- 15528009 TI - Effects of alcohol on behavioral and morphologic indices of sexual maturation in male rats. PMID- 15528010 TI - Influence of vitamin E, sodium selenite, and astrocyte-conditioned medium on neuronal survival after chronic exposure to ethanol. AB - Free radicals species generation during ethanol metabolism is implicated in ethanol-induced toxicity. Findings from clinical studies have clearly established the association between alcohol intake and nutritional deficiency. Astrocytes are able to promote neuronal survival against different lethal injuries involved in ethanol-induced toxicity. We therefore studied the ability of hydrosoluble vitamin E (trolox), sodium selenite, and astrocyte-conditioned medium to protect cultured rat neurones against ethanol-induced oxidative stress after chronic exposure to ethanol. When a 6-day exposure to ethanol (20 mM) led to a loss of cell viability, the presence of trolox (10 microM) offered a significant neuroprotection. In the presence of 3-amino-1,2,4-triazole, a catalase inhibitor that created conditions that were favorable to reactive oxygen species accumulation, trolox was able to counteract the deleterious effect of the inhibitor. Moreover, flow cytometric analysis indicated that trolox can maintain the intracellular glutathione content in neurones chronically exposed to ethanol. In these conditions of exposure, the absence of sodium selenite in the culture medium significantly aggravated the exposure-induced effects, whereas sodium selenite (100 nM) offered a significant neuroprotection. Finally, the presence of 25% astrocyte-conditioned medium in the neuronal culture medium induced a neuroprotective effect in the presence of ethanol. Nevertheless, when astrocytes were previously chronically (3 days) exposed to ethanol, their culture medium did not offer a significant protection. These results evidenced that vitamin E and astrocytes can protect neurones from ethanol-induced oxidative stress, notably by contributing to maintaining the intracellular glutathione levels. Selenium, by means of its exogenous addition in the form of sodium selenite, also had an interesting neuroprotective effect. PMID- 15528011 TI - Is maternal alcohol use a risk factor for early-onset sepsis in premature newborns? AB - Because chronic alcohol abuse alters immune defenses and increases infection in adults, we tested the hypothesis that maternal alcohol use during pregnancy would increase the risk of sepsis in very low birth weight (VLBW) premature newborns. We performed a case-controlled analysis of VLBW newborns born at Grady Memorial Hospital (Atlanta, GA). Alcohol exposure, as the predictive variable, was assessed by maternal self-report. The outcome variables were early-onset and multiple late-onset sepsis. Univariate analysis with Fisher exact test and multivariate analysis with the use of binary logistic regression were performed. Early-onset sepsis was 15-fold higher in the alcohol-exposed group (n=20) compared with findings for the matched control group (n=168) [alcohol-exposed group, 10%, vs. control group, 0.6%: odds ratio (OR) 6.8 (95% confidence interval [CI], 2.7-17.1), P < or = .05]. Early-onset sepsis in the alcohol+cocaine-exposed group (n=64) did not differ from findings for the control group. The prevalence of multiple late-onset sepsis did not differ among the exposure groups. Logistic regression analysis, controlling for chorioamnionitis and premature prolonged rupture of membranes, demonstrated an independent, increased risk of early-onset sepsis with alcohol exposure [OR 16 (95% CI, 1.2-210), P < or = .05]. We conclude that alcohol exposure significantly increased the risk of early-onset sepsis in this group of VLBW newborns. The effects of maternal alcohol abuse during pregnancy on the risk of infection in the VLBW newborn require further analysis. PMID- 15528012 TI - Ethanol suppresses cytokine responses induced through Toll-like receptors as well as innate resistance to Escherichia coli in a mouse model for binge drinking. AB - Toll-like receptors (TLRs) recognize molecular patterns associated with pathogens and initiate various mechanisms that are critical in innate resistance to infection. It has been reported that acute administration of ethanol suppresses responses mediated through TLR3 and TLR4. However, it is not known whether this is also true for other TLRs. Ligands for TLR2/TLR6 (zymosan A), TLR5 (bacterial flagellin), TLR7 (R-848), and TLR9 (CpG DNA) were used to induce cytokine production in mice, and the effects of ethanol (6 g/kg by gavage) on this induction were determined. Because different cell types may be affected differently by ethanol, cytokines were measured in serum (as an indication of cytokines produced by a number of different cell types) and in peritoneal lavage fluid (as an indicator of cytokine production primarily by peritoneal macrophages). Ethanol significantly affected the concentration of at least one of the cytokines evaluated in serum or peritoneal lavage fluid [interleukin (IL)-6, IL-10, and IL-12 p40 subunit] induced by all TLR ligands tested. The results also supported the suggestion that serum and peritoneal cytokines were mostly derived from different cells types, which were affected differently by ethanol. To determine whether ethanol-induced changes in TLR responses were associated with suppression of innate resistance to infection, a model of experimental peritonitis with a nonpathogenic (indigenous) strain of Escherichia coli was developed. Ethanol significantly decreased host resistance to E. coli peritonitis. Thus, ethanol suppresses responses induced by TLR receptors in mice and in the same experimental system it suppresses resistance to infection. PMID- 15528014 TI - Early events in acute pancreatitis. AB - Considerable progress in the understanding of the pathogenesis of acute pancreatitis is based on the conclusive finding that the initiation of the disease occurs within the acinar cell. Two lines of evidence have contributed to the progress in understanding the disease process: (1) the identification of patients with a hereditary form of pancreatitis as carriers of germline-mutations in the genes for cationic trypsinogen and the pancreatic secretory trypsin inhibitor and (2) the use of various transgenic and knock-out mouse strains in experimental models of acute pancreatitis. On the other hand, these studies have delivered several unexpected results that appear to be incompatible with long standing dogmas and paradigms of pancreatic research. Further progress in knowledge will result if the well-characterized enzymatic properties of human enzymes that are involved in the initial activation cascade can be investigated under in vivo conditions in transgenic animals or in permanent acinar cell lines. Such studies will permit the development of effective strategies for the prevention and treatment of this disease. PMID- 15528015 TI - Theories, mechanisms, and models of alcoholic chronic pancreatitis. AB - Alcoholic chronic pancreatitis is a severe, disabling, chronic inflammatory condition of the pancreas that is seen in fewer than 5% of alcoholics. The severity and unpredictability of this condition has lead to several theories on the mechanism causing chronic pancreatitis based on careful clinical observation. Hypothetical mechanisms were applied to various animal models. Finally, following multiple lines of evidence, there is a convergence of thought and development of some new models that are quite instructive. Taken together, chronic alcohol consumption by rats results in multiple effects on the pancreas that increase the risk of acute pancreatitis, including ongoing acinar cell injury that lowers the threshold for hyperstimulation-induced acute pancreatitis, neurohormonal injury, and adaptation that results in acinar cell hyperstimulation, increased susceptibility to viral mediated acute pancreatitis, and possibly other factors. After acute pancreatitis initiates the inflammatory process, the chronic inflammation and fibrosis of alcoholic chronic pancreatitis are driven by diet, the acinar cell stress response to continued alcohol that may be potentiated by toxic alcohol metabolites, hypoxia, hyperstimulation, and partial duct obstruction; plus the effects of proinflammatory immunocytes and cytokines; and by stellate cell-mediated fibrosis driven by anti-inflammatory cytokines, alcohol, and alcohol metabolites. The factors determining which alcoholic will develop alcoholic chronic pancreatitis likely involve genetic factors, dietary factors, and susceptibility to pancreatic injury through several mechanisms ranging from trauma to gallstones to viruses. PMID- 15528016 TI - Alcoholic pancreatitis. AB - Without doubt, alcohol consumption is one of the most important considerations in adults with acute or chronic pancreatitis. Understanding chronic pancreatitis as a complex disorder in which complimentary factors are required for recurrent acute and late chronic pancreatitis to develop in subsets of patients is critical for the early diagnosis and management of these individuals. Recent pathophysiological and genetic findings represent the beginning of major diagnostic and treatment breakthroughs that are likely to continue for the foreseeable future. The information provided in this article should provide the physician with a fresh perspective and remind the clinician of the importance of an accurate and complete history, and the need to document the actual alcohol consumption, pattern of drinking, and raise appropriate concerns if signs of alcoholism are detected. If alcohol-associated pancreatitis is detected, then limitation of pancreatic damage, limitation of progression, or preventative intervention should become the major concern. PMID- 15528017 TI - Cationic trypsinogen mutations and pancreatitis. AB - The discovery of PRSS1 mutations in hereditary pancreatitis and analysis of how the genotype affects the presentation and progression of hereditary pancreatitis has led to a better understanding of the pathophysiology of the disease. Patients with hereditary pancreatitis present with symptoms at an early age and have a significant lifetime risk for the development of endocrine and exocrine insufficiency, albeit at a later stage than patients with either idiopathic or alcoholic chronic pancreatitis. There are distinct phenotypic differences between hereditary pancreatitis and with other types of pancreatitis. As many as 80% of patients with symptomatic hereditary pancreatitis have an underlying causative PRSS1 mutation; there are, however, few significant phenotypic differences between these PRSS1 mutations. The R122H mutation is the most common PRSS1 mutation observed, and patients with the R122H mutation present earlier. This, however, does not necessarily translate into a more aggressive disease with respect to complications of chronic pancreatitis. Indeed, the age of presentation of symptoms may be a poor surrogate for predicting outcome, as inherited disorders of trypsinogen may cause subclinical attacks of pancreatitis, which ultimately lead to pancreatic destruction and dysfunction. All patients, irrespective of whether they carry a PRSS1 mutation, are at significant risk of developing pancreatic ductal adenocarcinoma. The risk appears to be insignificant below the age of 40 years, but it increases incrementally thereafter. Significantly, the risk of pancreatic cancer is not related to PRSS1 mutation type and does not appear to be related to the mode of inheritance. The role of SPINK1 mutations in modifying the expression of PRSS1 mutations is unclear but appears to be of clinical importance. It is unlikely that they act as causative mutations per se, at least in the Western form of the disease. Additionally, they do not appear to have an impact on the penetrance of PRSS1 gene mutations in hereditary pancreatitis. PMID- 15528018 TI - Serine protease inhibitor Kazal type 1 mutations and pancreatitis. AB - In summary, SPINK1 is thought to play an important role in protecting the pancreas against excessive trypsinogen activation. SPINK1 mutations are associated with the development of acute and chronic pancreatitis and have been detected in all forms of chronic pancreatitis. The strong association of mutations in the PRSS1 gene and in the SPINK1 gene with chronic pancreatitis supports the concept of intracellular trypsin activation as an initiating and extremely important step in the development of pancreatitis. The N34S mutation represents the most frequently observed pancreatitis-associated SPINK1 variant. Because the SPINK1 N34S mutation is very common in the general population, it is unlikely that this mutation alone can initiate the development of chronic pancreatitis. Thus, it rather appears that in most patients with SPINK1 associated chronic pancreatitis, this genetic variant acts as disease modifier or within a polygenic model with other yet unidentified genes or environmental cofactors. The possible interaction of mutations in the SPINK1 gene with other pancreatitis-associated susceptibility genes has to be investigated in future research efforts. The most promising candidate gene for such an interaction is the CFTR gene, because genetic alterations within the CFTR gene are also common in the general population and already have been associated with chronic pancreatitis. PMID- 15528019 TI - Susceptibility to pancreatitis related to PSTI/SPINK1 expression. AB - This article summarized several observations on the role of pancreatic secretory trypsin inhibitor in the pancreas. Although it long has been suspected that endogenous pancreatic trypsin inhibitors protect against inadvertent activation of trypsinogen, this hypothesis has gained strength from recent biochemical investigations and genetic studies of populations suffering from chronic pancreatitis. There is now considerable evidence from clinical disease associations and burgeoning experimental models that some forms of pancreatitis may be the result of an imbalance between active pancreatic proteases and their inhibitors within the pancreas. Future studies should clarify the precise molecular interactions between enzymes and inhibitors and how these may be manipulated to prevent or treat pancreatitis. PMID- 15528020 TI - The role of cystic fibrosis gene mutations in determining susceptibility to chronic pancreatitis. AB - This article reviews current concepts regarding the pathobiology of cystic fibrosis pancreatic disease. It summarizes recent studies on the relationship between CFTR mutations and pancreatitis, and it reviews several unresolved issues in the field. PMID- 15528021 TI - Genetic counseling for hereditary pancreatitis--the role of molecular genetics testing for the cationic trypsinogen gene, cystic fibrosis and serine protease inhibitor Kazal type 1. AB - The importance of pretest information, using an accredited DNA laboratory and interpreting the genotype on behalf of the patient and their physicians is emphasized. Care with predictive testing and the strong encouragement to involve a specialist genetic counseling service is made. A similar approach to genetic testing should be used when children are involved. Because of the incomplete pickup of PRSS1 mutations, particularly of a limited mutation panel of R122H and N291 (perhaps with A16V), a diagnosis of HP cannot be ruled out by molecular genetic testing alone. The A16V mutation has a reduced penetrance, and its contribution to pancreatitis remains unclear. The advice to patients with genetic forms of pancreatitis is a strong encouragement to avoid smoking, to avoid alcohol, and to remain in contact with clinical and research groups for their follow-up and screening trials for early pancreatic cancer. The remaining issues are of how wide to cast the net of investigation in patients with unexplained pancreatitis, particularly looking for mutations in the CFTR and lower penetrance genes such as PSTI/SPINK1. PMID- 15528022 TI - Medical treatment of acute pancreatitis. AB - Eighty percent of all cases of acute pancreatitis are linked etiologically to gallstone disease or caused by immoderate alcohol consumption. No specific causal treatment for acute pancreatitis exists. Early prognostic factors that indicate severe disease are three or more signs on organ failure scores according to Ranson, Imrie, or Acute Physiology and Chronic Health Evaluation (APACHE) 11, extrapancreatic complications of the disease, or the detection of pancreatic necrosis on CT scans. Elevated CRP levels above 130 mg/L can also predict a severe course of acute pancreatitis. The essential medical treatment for acute pancreatitis is the correction of hypovolemia. Moreover, relief of often severe visceral pain is a high priority. Prophylactic antibiotics should be restricted to patients with necrotizing pancreatitis, infected necrosis, or other infectious complications. Enteral nutrition has no adverse effect compared with parenteral nutrition during the course of acute pancreatitis, and is probably beneficial in regard to outcome. PMID- 15528023 TI - Predictors of severity and necrosis in acute pancreatitis. AB - C-reactive protein remains the single standard biochemical marker for predicting the severity of AP. Because the combination of clinical-physiological scores and CRP provide good information at 48 hours, research has focused on the predictive ability of various markers when applied in the initial 24 hours after admission to the hospital. After detailed review of the literature, the authors conclude that there is no single tool that serves as the optimal predictor of severity. There are, however, data that support the use of certain tests to improve upon the clinician's early predictive ability on the subsequent course of AP. These include an APACHE II score greater than 7 and IL-6 at the time of admission, and urine TAP, urine trypsinogen-2, and serum PMN elastase at 24 hours (Table 4). These markers only will be able to help the clinician's predictive ability if they can be performed locally and if the results can be available ina timely manner. Future research should focus on promising markers such as procalcitonin, IL-8, IL-I ra, sTNFR, CAPAP, PLA-2, novel markers, and the combined use of more than one marker. The conventional research approach in predicting severity used in the last 15 years has limitations and appears to have reached its maximal potential. Novel conceptions and approaches, such as identification of genetic polymorphisms that predispose to severe course and complications of AP or other approaches are needed for a quantum step forward. PMID- 15528024 TI - Molecular diagnosis of solid and cystic lesions of the pancreas. AB - Pancreatic cancer presents at a clinically advanced stage. Diagnosis often is delayed in those who undergo several negative investigations, and at times, the diagnosis ultimately is made at surgery. Tumor suppressor-linked microsatellite allelic loss analysis appears to improve the diagnostic accuracy of inconclusive cytology specimens. Science, however, still suffers from the lack of a reliable biomarker for the purpose of screening and early detection. The continued study of sera and pancreatic juice hopefully will lead to the discovery of molecular biomarkers capable of diagnosing pre-clinical disease in at-risk groups. The ability to predict the underlying biologic behavior of pancreatic cystic neoplasia is less than ideal. Tumor suppressor-linked microsatellite allelic loss and telomerase analysis show promise, but they have not been studied extensively. Proteomic analysis of pancreatic cyst fluid, although not yet reported, also may prove valuable in guiding clinical management. PMID- 15528025 TI - Identification of high-risk pancreatic cancer-prone families. AB - Pancreatic cancer-prone families provide a unique resource for studying the etiology, natural history, genetics, and treatment of pancreatic cancer. The only effective way of identifying these families is by obtaining a complete family history, since it is not possible to differentiate sporadic pancreatic cancer cases from hereditary cases based on either clinical presentation or features. These families also would benefit greatly from early detection or prevention strategies. Ultimately, this knowledge could be applied to the more common sporadic form of pancreatic cancer, where diagnosis is almost always late, and prognosis remains quite grim. PMID- 15528026 TI - Genetics of hereditary pancreatic carcinoma. AB - Several tumor syndromes are known to have an increased incidence of pancreatic carcinoma. Unfortunately, the incidence of pancreatic carcinoma almost equals its mortality. Because of intense research activities in the past decade, several genetic alterations have been identified that are responsible for the so-called sporadic pancreatic carcinoma development. National and international research activities only recently have begun focusing on the molecular pathogenesis of hereditary risk factors for pancreatic carcinoma. It is hoped that these genetic studies will aid the development of clinical management strategies for pancreatic carcinoma prevention. PMID- 15528027 TI - Flavonoids and the brain: interactions at the blood-brain barrier and their physiological effects on the central nervous system. AB - Over the past few years there has been an exponential growth in the number of reports describing the effects of nutritional modulation on aging and age-related diseases. Specific attention has been directed toward the beneficial effects afforded by dietary antioxidants, in particular those from fruit and vegetables, in ameliorating age-related deficits in brain performance. The rationale for studying the effects of dietary intervention stems from evidence implicating free radicals in aspects related to the aging process. Age-dependent neuropathology is a cumulative response to alterations induced by reactive oxygen species. Therefore cognitive aging, according to this hypothesis, should be slowed, and possibly even reversed, by appropriately increasing levels of antioxidants or decreasing overproduction of free radicals in the body. PMID- 15528028 TI - Signaling kinases modulated by 4-hydroxynonenal. AB - The interaction of 4-hydroxynonenal (HNE) with a variety of kinases variously involved in cell signaling is now a matter of active investigation. In particular, findings with regard to the effect of HNE on different components of the protein kinase C family and the mitogen-activated protein kinase complex already provide reliable indications of a potential role of this aldehyde as a cell signal messenger. Such a role appears further supported by the clear-cut evidence of up-regulation of receptor tyrosine kinases and down-regulation of the nuclear factor kappa B system, produced by HNE concentrations actually detectable in pathophysiology. PMID- 15528029 TI - The peroxynitrite pathway in development: phenytoin and benzo[a]pyrene embryopathies in inducible nitric oxide synthase knockout mice. AB - Nitric oxide generated by nitric oxide synthases (NOSs) can react with reactive oxygen species (ROS), forming peroxynitrite, which may contribute to the ROS initiated macromolecular damage implicated in the embryopathic effects of both endogenous and drug-enhanced oxidative stress. Inducible NOS (iNOS) is nonconstitutive in most tissues, and its embryonic expression and developmental importance are unknown. Herein, during organogenesis (Gestational Days 9 and 10), wild-type B6129PF2 embryos in culture were highly susceptible to the ROS initiating teratogens phenytoin and benzo[a]pyrene, whereas iNOS knockout embryos were substantially but not completely protected (p < .05), implicating iNOS in the embryopathic mechanism. However, in contrast to prostaglandin H synthase catalyzed teratogen bioactivation and ROS formation, which occurs within the embryo, in vivo iNOS expression was limited to placental tissue. These results suggest that the diffusion of nitric oxide from placental progenitor tissue (ectoplacental cone) to embryonic target tissues contributes to the embryopathic effects of ROS-initiating teratogens in embryo culture, which may constitute a mechanism by which embryonic determinants of ROS-mediated teratogenesis can be modulated by maternal extra-embryonic processes. PMID- 15528030 TI - Enrichment of rat hepatic organelles by vitamin E administered subcutaneously. AB - Novel modes of administering antioxidants to improve delivery to targeted tissues or cells may be advantageous in preventing oxidant-induced pathologies. Vitamin E (alpha-tocopherol) has been shown to be protective in several models of liver injury. The objectives of this study were: (1) to determine if subcutaneously (s.q.) administered emulsified vitamin E enriched liver and hepatic subcellular fractions with the antioxidant and (2) to carry out a time-dependent analysis of serum and tissue vitamin E in rats receiving daily s.q. vitamin E. In the first experiment rats injected daily s.q. with emulsified vitamin E for 9 d increased serum, total liver, liver mitochondria, and liver microsomes by 8-, 16-, 30-, and 29-fold, respectively, compared with placebo injections. Similar enrichment was observed after intramuscular injections. In the second experiment, daily doses of s.q. vitamin E increased liver concentrations 40-fold by 9 d, which decreased to 22-fold by 18 d, whereas serum adjusted vitamin E levels maximized with a 24-fold increase by day 3 and plateaued thereafter. In conclusion, s.q. administration of emulsified vitamin E to rats resulted in substantially elevated serum and liver concentrations of alpha-tocopherol compared with levels achievable by dietary supplementation. The s.q. route of administration is a potentially effective parenteral mode of delivery of vitamin E for conditions in which hepatic oxidative stress is present. PMID- 15528031 TI - Ethacrynic-acid-induced glutathione depletion and oxidative stress in normal and Mrp2-deficient rat liver. AB - Oxidative stress in the liver is sometimes accompanied by cholestasis. We investigated the localization and role of multidrug-resistance-associated protein (Mrp) 2, a biliary transporter involved in bile-salt-independent bile flow, under ethacrynic acid (EA)-induced acute oxidative stress. Normal Sprague-Dawley rat (SDR) and Mrp2-deficient Eisai hyperbilirubinemic rat (EHBR) livers were perfused with 500 microM EA. The release of glutamic pyruvic transaminase (GPT) and thiobarbituric-acid-reactive substances (TBARS) from EHBR liver was markedly delayed compared with that from SDR liver. This is mainly due to the higher basal level of glutathione (GSH) in EHBR liver (59.1 +/- 0.3 nmol/mg protein) compared with SDR liver (39.7 +/- 1.5 nmol/mg protein). EA similarly induced a rapid reduction in GSH followed by mitochondrial permeability transition in the isolated mitochondria from both SDR and EHBR. Internalization of Mrp2 was detected before nonspecific disruption of the canalicular membrane and GPT release in SDR liver perfused with 100 microM EA. SDR liver preperfused with hyperosmolar buffer (405 mosmol/L) for 30 min induced internalization of Mrp2 without changing the basal GSH level, while elimination of hepatic GSH by 300 microM EA perfusion was significantly delayed thereafter. Concomitantly, hepatotoxicity assessed by the release of GPT and TBARS was also significantly attenuated under hyperosmolar conditions. In conclusion, preserved cytosolic and intramitochondrial GSH is the key factor involved in the acute hepatotoxicity induced by EA and its susceptibility could be altered by the presence of Mrp2. PMID- 15528032 TI - Protective effects of 17beta-estradiol and trivalent chromium on interleukin-6 secretion, oxidative stress, and adhesion of monocytes: relevance to heart disease in postmenopausal women. AB - Postmenopausal diabetic women are at greater risk for heart disease compared with men of similar age and with other risk factors. We examined the hypothesis that 17beta-estradiol and trivalent chromium inhibit secretion of the pro-inflammatory cytokine interleukin (IL)-6 and oxidative stress in monocytes exposed to high glucose (HG). U937 human monocytes were cultured with HG (30 mM) with and without 17beta-estradiol (0-1000 nM) and chromium chloride (Cr(3+), 0-10 muM) at 37 degrees C for 24 h. Results show that 17beta-estradiol inhibits IL-6 and adhesion to endothelial cells (p <. 05) by HG-treated monocytes. Treatment with 17beta estradiol+Cr(3+) required a significantly lower dose of estradiol-17beta compared with 17beta-estradiol alone for IL-6 inhibition. 17beta-Estradiol+Cr(3+) also inhibited lipid peroxidation and the adhesivity to human endothelial cells in HG treated monocytes. Thus, 17beta-estradiol+Cr(3+) inhibits oxidative stress, IL-6 secretion, and monocytic adhesion to endothelial cells, risk factors in the development of heart disease. The female body requires E but studies on some patients indicate side effects with increased amounts of 17beta-estradiol supplementation. The potential benefit of a lower estrogen dose in combination with chromium is novel and needs to be explored in postmenopausal diabetic women. PMID- 15528033 TI - Lung injury and mortality with hyperoxia are increased in peroxiredoxin 6 gene targeted mice. AB - Overexpression of peroxiredoxin 6 (Prdx6) has been shown to protect lungs of mice against hyperoxia-mediated injury. In this study, we evaluated whether genetic inactivation of Prdx6 in mice increases sensitivity to oxygen toxicity. We evaluated mouse survival, lung histopathology, total protein and nucleated cells in bronchoalveolar lavage fluid (BALF), and oxidation of lung protein and lipids by measurement of protein carbonyls and thiobarbituric reactive substances (TBARS), respectively. The duration of survival for Prdx6 -/- mice was significantly shorter than that observed in wild-type mice on exposure to 85 or 100% O(2); survival of Prdx6 +/- mice was intermediate. After 72-h exposure to 100% O(2), lungs of Prdx6-/- mice showed more severe injury than wild-type with increased wet/dry weight, epithelial cell necrosis and alveolar edema on microscopic examination, increased protein and nucleated cells in BALF, and higher content of TBARS and protein carbonyls in lung homogenate. These findings show that Prdx6 -/- mice have increased sensitivity to hyperoxia and provide in vivo evidence that Prdx6 is an important lung antioxidant enzyme. PMID- 15528034 TI - Hypoxia/reoxygenation and vitamin C intake influence NO synthesis and antioxidant defenses of neutrophils. AB - Oxidative stress induced by hypoxia/reoxygenation mediates the pathophysiological consequence of ischemia/reperfusion and human diseases. Diving apnea could be a good model of oxidative stress induced by hypoxia/reoxygenation. We studied the influence of vitamin C diet supplementation on the response of neutrophil antioxidant defenses, NO production, and redox status to diving apnea. Seven professional apnea divers participated in a double-blind cross study. Divers were assigned to either vitamin C-supplemented (1 g/d for a week) or placebo groups. Blood samples were taken under basal conditions, immediately after diving apnea for 4 h and after 1 h of recovery. Plasma vitamin C increased only in the supplemented group after diving and was maintained high in recovery. Diving apnea decreased neutrophil GSH/GSSG ratio in both groups, but maintained protein carbonyl derivates. Neutrophil catalase activity and levels and glutathione peroxidase activity were lower in the supplemented group than in the placebo group after diving. iNOS and nitrite levels decreased only in the supplemented group after diving and recovery. Diving apnea induced oxidative stress and initiated neutrophil reactions that resemble the acute-phase immune response with increased myeloperoxidase activity in neutrophils. Diet supplementation with vitamin C reduced neutrophil iNOS levels and NO production. PMID- 15528035 TI - Biosynthesis and turnover of DOPA-containing proteins by human cells. AB - Protein-bound 3,4-dihydroxyphenylalanine (PB-DOPA) is a major product of hydroxyl radical attack on tyrosine residues of proteins. Levels of PB-DOPA in cells and tissues have been shown to be greatly elevated in age-related diseases. We demonstrate for the first time that l-DOPA (levodopa) can be biosynthetically incorporated into cell proteins by human cells (THP-1 monocytes and monocyte derived macrophages). The DOPA-containing proteins generated were selectively visualized on PVDF membranes using a redox-cycling staining method. Many cell proteins contained DOPA and seemed to be synthesized as their full-length forms. The cellular removal of DOPA-containing proteins by THP-1 cells was by proteolysis involving both the proteasomal and the lysosomal systems. The rate of cellular proteolysis of DOPA-containing proteins increased at lower levels of DOPA incorporation but decreased at higher levels of DOPA incorporation. The decreased rate of degradation was accompanied by an increase in the activity of cathepsins B and L but the activity of cathepsin S increased only at lower levels of DOPA incorporation. These data raise the possibility that PB-DOPA could be generated in vivo from l-DOPA, which is the most widely used treatment for Parkinson disease. PMID- 15528036 TI - Glycation-induced inactivation of NADP(+)-dependent isocitrate dehydrogenase: implications for diabetes and aging. AB - Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and the cellular defense against oxidative damage is one of the primary functions of NADP(+)-dependent isocitrate dehydrogenase (ICDH), because it supplies NADPH for antioxidant systems. When exposed to reducing sugars such as glucose, glucose 6-phosphate, and fructose, ICDH was susceptible to oxidative modification and damage, which was indicated by a loss of activity and fragmentation of the peptide as well as by the formation of carbonyl groups. The glycated ICDH was isolated and identified by boronate-affinity chromatography and immunoblotting with anti-hexitol-lysine antibody. The active site lysine residue, Lys(212), was identified as one of the major sites of nonenzymatic glycation of ICDH. The structural alterations of modified enzymes were indicated by changes in thermal stability, intrinsic tryptophan fluorescence, and binding of the hydrophobic probe 8-anilino-1-naphthalene sulfonic acid. When we examined the antioxidant role of mitochondrial ICDH against glycation-induced cytotoxicity with HEK293 cells transfected with the cDNA for mouse mitochondrial ICDH in sense and antisense orientations, a clear inverse relationship was observed between the amount of mitochondrial ICDH expressed in target cells and their susceptibility to glycation-mediated cytotoxicity. Mitochondrial ICDH was purified by immunoprecipitation and probed with anti-hexitol-lysine antibody, which revealed increased levels of glycated ICDH in the kidneys of diabetic rats and in the lenses of diabetic patients suffering from cataracts. A decrease in ICDH activity was observed in those tissues. We also found that levels of glycated ICDH increased in IMR-90 cells and rat kidney during normal aging. The glycation mediated damage to ICDH may result in the perturbation of cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant condition and may contribute to various pathologies associated with the general aging process and long-term complications of diabetes. PMID- 15528037 TI - alpha-keto-beta-methyl-n-valeric acid diminishes reactive oxygen species and alters endoplasmic reticulum Ca(2+) stores. AB - Mitochondrial dysfunction and oxidative stress occur in neurodegenerative diseases. Other results show that bombesin-releasable calcium stores (BRCS) from the endoplasmic reticulum (ER) are exaggerated in fibroblasts from patients with Alzheimer's disease (AD) compared with controls and in fibroblasts from a young control treated with H(2)O(2). We hypothesize that alterations in oxidative stress underlie the exaggeration in BRCS in AD, and that appropriate antioxidants may be useful in treating this abnormality. Two indicators of different oxidant species were used to determine the effects of select oxidants on cellular oxidation status: carboxydichlorofluorescein (c-DCF) to detect reactive oxygen species (ROS), and 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF) to detect nitric oxide (NO(.-)). Various conditions that induce ROS, including H(2)O(2), oxygen/glucose deprivation, and 3-morpholinosyndnonimine (SIN-1), were used to test the ability of alpha-keto-ss-methyl-n-valeric acid (KMV) to scavenge ROS. KMV diminished c-DCF-detectable ROS that were induced by H(2)O(2), oxygen/glucose deprivation, or SIN-1 in PC12 cells, primary neuronal cultures, or fibroblasts. Furthermore, KMV reduced the H(2)O(2)-induced increase in BRCS and diminished the elevation in BRCS in cells from AD patients to control levels. On the other hand, DAF-detectable NO(.-) induced by SIN-1 was not scavenged by KMV and did not exaggerate BRCS. The results indicate that KMV is an effective antioxidant of c-DCF-detectable ROS. The effects of KMV are not cell type specific, but are ROS specific. The same H(2)O(2)-induced ROS that reacts with KMV may also underlie the changes in BRCS related to AD. Thus, KMV ameliorates the effects of ROS on calcium homeostasis related to oxidative stress and to AD. PMID- 15528038 TI - Protective effect of melatonin on beta-amyloid-induced apoptosis in rat astroglioma C6 cells and its mechanism. AB - Astrocytosis is a common feature of amyloid plaques. The Abeta-astrocyte interaction produces a detrimental effect on neurons, which may contribute to neurodegeneration in Alzheimer disease (AD). The regulation of astrocyte apoptosis is essential to physiological and pathological processes in the CNS. Melatonin is a potent antioxidant and free radical scavenger. Previously, we showed that melatonin alleviated the learning and memory deficits in the APP 695 transgenic mouse model of AD. In this study, the importance of melatonin in the management of Abeta-induced apoptosis in an astrocyte-like cell is discussed. We found that rat astroglioma C6 cells treated with Abeta25-35 or Abeta1-42 undergo apoptosis and that melatonin pretreatment at 10(-5), 10(-6), and 10(-7) M significantly attenuates Abeta25-35- or Abeta1-42-induced apoptosis. The antiapoptotic effects of melatonin were extremely reproducible and corroborated by multiple quantitative methods, including an MTT cell viability assay, Hoechst 33342 nuclei staining, DNA fragmentation analysis, and flow cytometric analysis. In addition, melatonin effectively suppressed Abeta1-42-induced nitric oxide formation, remarkably prevented Abeta1-40-induced intracellular calcium overload, and significantly alleviated Abeta1-40-induced membrane rigidity. Our results demonstrate that, in addition to the beneficial effects of providing direct antioxidant protection to neurons, melatonin may enhance neuroprotection against Abeta-induced neurotoxicity by promoting the survival of glial cells. PMID- 15528039 TI - Carbon monoxide actuates O(2)-limited heme degradation in the rat brain. AB - The biochemical paradigm for carbon monoxide (CO) is driven by the century-old Warburg hypothesis: CO alters O(2)-dependent functions by binding heme proteins in competitive relation to 1/oxygen partial pressure (PO(2)). High PO(2) thus hastens CO elimination and toxicity resolution, but with more O(2), CO-exposed tissues paradoxically experience less oxidative stress. To help resolve this paradox we tested the Warburg hypothesis using a highly sensitive gas-reduction method to track CO uptake and elimination in brain, heart, and skeletal muscle in situ during and after exogenous CO administration. We found that CO administration does increase tissue CO concentration, but not in strict relation to 1/PO(2). Tissue gas uptake and elimination lag behind blood CO as predicted, but 1/PO(2) vs. [CO] fails even at hyperbaric PO(2). Mechanistically, we established in the brain that cytosol heme concentration increases 10-fold after CO exposure, which sustains intracellular CO content by providing substrate for heme oxygenase (HO) activated after hypoxia when O(2) is resupplied to cells rich in reduced pyridine nucleotides. We further demonstrate by analysis of CO production rates that this heme stress is not due to HO inhibition and that heme accumulation is facilitated by low brain PO(2). The latter becomes rate limiting for HO activity even at physiological PO(2), and the heme stress leads to doubling of brain HO-1 protein. We thus reveal novel biochemical actions of both CO and O(2) that must be accounted for when evaluating oxidative stress and biological signaling by these gases. PMID- 15528040 TI - Effects of thiol antioxidant on reduced nicotinamide adenine dinucleotide phosphate oxidase in hypertensive Dahl salt-sensitive rats. AB - Recent studies implicate of reactive oxygen species (ROS) in hypertension; however, whether reactive oxygen species promote hypertensive derangements is not fully clear. We thus investigated the effects of an antioxidant, N-acetyl-L cysteine, on hypertensive Dahl salt-sensitive rats. High-salt intake for 4 weeks markedly elevated systolic arterial pressure, urinary excretion of protein, 8 isoprostane, and H(2)O(2), and the enzyme activity of reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase along with the elevated expression of its subunits gp91phox and p47phox at the levels of mRNA and protein. Supplement with N-acetyl-L-cysteine reduced the increase in systolic arterial pressure and counteracted the elevation of urinary excretion of protein, 8 isoprostane, and H(2)O(2), and the increases in NADPH oxidase activity/expression in high-salt-loaded Dahl salt-sensitive rats. N-acetyl-L-cysteine supplement ameliorated plasma and urinary levels of thromboxane B(2) (an end metabolite of thromboxane A(2)), associated with improvement of both the abnormal contraction and the impaired nitric oxide-dependent relaxation in renal arteries. These results revealed that oxidative stress mediates hypertensive changes in Dahl salt sensitive rats, because thiol antioxidant N-acetyl-L-cysteine attenuated the augmentation of local ROS production by diminishing the elevation of NADPH oxidase expression and ameliorated renal/vascular hypertensive changes. PMID- 15528041 TI - Oxidative damage during chagasic cardiomyopathy development: role of mitochondrial oxidant release and inefficient antioxidant defense. AB - In this study, we evaluated the oxidant status and antioxidant defense capabilities of the heart during the course of Trypanosoma cruzi infection and disease development in a murine model system. Our data show that the extent of protein carbonylation and lipid peroxidation is increased in the heart, but not the skeletal muscle, of infected mice. The level of oxidative injury biomarkers in the myocardium consistently increased with chronic disease severity. The antioxidant defense constituted by catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GSR), and reduced glutathione was increased in murine heart and skeletal tissue in response to the stress of T. cruzi infection. After the initial burst, CAT, GPx, and GSR remained unresponsive to the severity of chronic tissue damage in chagasic hearts. The cardiac level of Mn(2+) superoxide dismutase (MnSOD) was diminished in chagasic mice. Our data suggest that the host responds to acute injuries by activating antioxidant defenses that are of sufficient magnitude to scavenge the reactive oxidants in skeletal tissue. The myocardia of infected mice, however, sustain increased oxidative injuries with disease progression. We surmise that MnSOD deficiencies, resulting in the increased release of mitochondrial free radicals, lead to sustained oxidative stress that exceeds the cardiac antioxidant defense capacity and contribute to persistent oxidative damage in chagasic myocardium. PMID- 15528042 TI - Dual role of cAMP in iNOS expression in glial cells and macrophages is mediated by differential regulation of p38-MAPK/ATF-2 activation and iNOS stability. AB - We reported previously that cAMP analogues or cAMP synthesis activator (forskolin; FSK) inhibit lipopolysaccharide (LPS)-induced inducible nitric-oxide systase (iNOS) gene expression in astrocytes, while they enhance that in macrophages. Here, we report that the FSK-mediated inhibition of iNOS expression in C6 glial cells is due to its reduced transcriptional activity, while the FSK mediated enhancement of iNOS expression in RAW264.7 macrophages is a result of increased stability of iNOS protein without transcriptional enhancement. The LPS/interferon-gamma (IFN)-induced iNOS transcription was inhibited by FSK via inhibition of p38-MAPK/ATF-2 activity in glial cells while it was not affected in macrophages. In both cell types, proteasome activities were required for the spontaneous degradation of iNOS protein, and the inhibition of proteasome activity by MG132 after maximum increase of iNOS protein levels further enhanced iNOS protein induction by LPS/IFN, suggesting the involvement of proteasome in iNOS degradation. More importantly, the iNOS protein levels were equalized by the MG132 posttreatment in macrophages treated with LPS/IFN alone and along with FSK, and ubiquitinated iNOS protein levels were reduced by FSK posttreatment, suggesting that the FSK-mediated inhibition of ubiquitination of iNOS protein and the following increased stability of iNOS protein are one of the mechanisms of cAMP-pathway-mediated enhancement of iNOS gene expression in macrophages. To our knowledge, this is the first evidence that cAMP regulates iNOS expression at the posttranslational level in macrophages. PMID- 15528043 TI - Bcl-2 family proteins regulate mitochondrial reactive oxygen production and protect against oxidative stress. AB - Bcl-2 family proteins protect against a variety of forms of cell death, including acute oxidative stress. Previous studies have shown that overexpression of the antiapoptotic protein Bcl-2 increases cellular redox capacity. Here we report that cell lines transfected with Bcl-2 paradoxically exhibit increased rates of mitochondrial H(2)O(2) generation. Using isolated mitochondria, we determined that increased H(2)O(2) release results from the oxidation of reduced nicotinamide adenine dinucleotide-linked substrates. Antiapoptotic Bcl-2 family proteins Bcl-xL and Mcl-1 also increase mitochondrial H(2)O(2) release when overexpressed. Chronic exposure of cells to low levels of the mitochondrial uncoupler carbonyl cyanide 4-(triflouromethoxy)phenylhydrazone reduced the rate of H(2)O(2) production by Bcl-xL overexpressing cells, resulting in a decreased ability to remove exogenous H(2)O(2) and enhanced cell death under conditions of acute oxidative stress. Our results indicate that chronic and mild elevations in H(2)O(2) release from Bcl-2, Bcl-xL, and Mcl-1 overexpressing mitochondria lead to enhanced cellular antioxidant defense and protection against death caused by acute oxidative stress. PMID- 15528044 TI - Participation of singlet oxygen in ultraviolet-a-induced lipid peroxidation in mouse skin and its inhibition by dietary beta-carotene: an ex vivo study. AB - Dietary beta-carotene acts as a photoprotective agent in the skin, but the exact mechanism of protection is unknown. This ex vivo study is focused on determining the mechanism of action of beta-carotene against UV-A-induced skin damage by characterizing peroxidized phosphatidylcholine (PC) and beta-carotene oxidation products. BALB/c mice were fed with basal or a beta-carotene-supplemented diet, and homogenates from their dorsal skin were prepared after 3 weeks for UV-A irradiation. Analyses revealed that the degree of lipid peroxidation in the beta carotene group was significantly lower than that in the controls. The isomeric composition of hydroperoxy fatty acids, constituting peroxidized PC, was determined by thin-layer chromatography-blotting followed by gas chromatography/mass spectrometry (MS)/selected ion monitoring analysis. The 9- and 10-isomers of peroxidized PC, resulting from the reaction of singlet molecular oxygen ((1)O(2)) with oleic acid, were elevated in the UV-A-exposed control group compared to the experimental group. Similar results were obtained from methylene-blue-sensitized photooxidation of mouse skin lipids in vitro. Liquid chromatography/MS analysis of the homogenates confirmed the formation of beta-carotene 5,8-endoperoxide, a specific marker for the (1)O(2) reaction. These results indicate that dietary beta-carotene accumulates in the skin and acts as a protective agent against UV-A-induced oxidative damage, by quenching the (1)O(2). PMID- 15528045 TI - Identification and quantification of N(epsilon)-(Hexanoyl)lysine in human urine by liquid chromatography/tandem mass spectrometry. AB - The identification and quantification of N(epsilon)-(hexanoyl)lysine (N(epsilon) HEL), which was found from the reactions between lipid hydroperoxide and lysine, from human urine was examined using liquid chromatography/tandem mass spectrometry (LC/MS/MS). The N(epsilon)-HEL in the partially purified urine fraction was identified using LC/MS/MS by several approaches including precursor/product ion scans. The peak found by the multiple-reaction monitoring (MRM) of the collision-induced fragmentation of N(epsilon)-HEL was clearly observed in urine, and the elution position coincided with the synthetic standard N(epsilon)-HEL. The product, estimated N(epsilon)-HEL, was absorbed by a specific antibody to N(epsilon)-HEL. Moreover, N(alpha)-HEL, one of the plausible hexanoyl adducts from the reaction between the N(alpha) moiety of L-lysine and the peroxidized lipid, was hardly detected in urine samples, suggesting that the origin of the N(epsilon)-HEL is the peroxidized lipid-modified proteins but not artificial hexanoylated L-lysine. Using the MRM technique, the amount of urinary N(epsilon)-HEL from the control subjects (observed healthy) was estimated to be 1.58 +/- 0.23 mumol/mol of creatinine. A comparative study of the urinary N(epsilon)-HEL with an oxidative stress marker, 8-oxo-7,8-dihydro-2' deoxyguanosine, showed a high correlation (r = 0.844) between the two biomarkers. Furthermore, the quantification of N(epsilon)-HEL in the control and diabetic urines revealed that the urinary N(epsilon)-HEL from diabetic subjects (3.21 +/- 0.65 mumol/mol of creatinine) was significantly higher than that from the control subjects. PMID- 15528046 TI - Nuclear glutathione S-transferase pi prevents apoptosis by reducing the oxidative stress-induced formation of exocyclic DNA products. AB - We previously found that nuclear glutathione S-transferase pi (GSTpi) accumulates in cancer cells resistant to anticancer drugs, suggesting that it has a role in the acquisition of resistance to anticancer drugs. In the present study, the effect of oxidative stress on the nuclear translocation of GSTpi and its role in the protection of DNA from damage were investigated. In human colonic cancer HCT8 cells, the hydrogen peroxide (H(2)O(2))-induced increase in nuclear condensation, the population of sub-G(1) peak, and the number of TUNEL-positive cells were observed in cells pretreated with edible mushroom lectin, an inhibitor of the nuclear transport of GSTpi. The DNA damage and the formation of lipid peroxide were dependent on the dose of H(2)O(2) and the incubation time. Immunological analysis showed that H(2)O(2) induced the nuclear accumulation of GSTpi but not of glutathione peroxidase. Formation of the 7-(2-oxo-hepyl)-substituted 1,N(2) etheno-2'-deoxyguanosine adduct by the reaction of 13-hydroperoxyoctadecadienoic acid (13-HPODE) with 2'-deoxyguanosine was inhibited by GSTpi in the presence of glutathione. The conjugation product of 4-oxo-2-nonenal, a lipid aldehyde of 13 HPODE, with GSH in the presence of GSTpi, was identified by LS/MS. These results suggested that nuclear GSTpi prevents H(2)O(2)-induced DNA damage by scavenging the formation of lipid-peroxide-modified DNA. PMID- 15528047 TI - Redox-dependent effects of nitric oxide on microvascular integrity in oxygen induced retinopathy. AB - Opposing effects have been ascribed to nitric oxide (NO) on retinal microvascular survival. We investigated whether changes in the redox state may contribute to explain apparent conflicting actions of NO in a model of oxygen-induced retinal vasoobliteration. Retinal microvascular obliteration was induced by exposing 7 day-old rat pups (P7) for 2 or 5 days to 80% O(2). The redox state of the retina was assessed by measuring reduced glutathione and oxidative and nitrosative products malondialdehyde and nitrotyrosine. The role of NO on vasoobliteration was evaluated by treating animals with nitric oxide synthase (NOS) inhibitors (N nitro-l-arginine; L-NA) and by determining NOS isoform expression and activity; the contribution of nitrosative stress was also determined in animals treated with the degradation catalyst of peroxynitrite FeTPPS or with the superoxide dismutase mimetic CuDIPS. eNOS, but not nNOS or iNOS, expression and activity were increased throughout the exposure to hyperoxia. These changes were associated with an early (2 days hyperoxia) decrease in reduced glutathione and increases in malondialdehyde and nitrotyrosine. CuDIPS, FeTPPS, and L-NA treatments for these 2 days of hyperoxia nearly abolished the vasoobliteration. In contrast, during 5 days exposure to hyperoxia when the redox state rebalanced, L-NA treatment aggravated the vasoobliteration. Interestingly, VEGFR-2 expression was respectively increased by NOS inhibition after short-term (2 days) exposure to hyperoxia and decreased during the longer hyperoxia exposure. Data disclose that the dual effects of NO on newborn retinal microvascular integrity in response to hyperoxia in vivo depend on the redox state and seem mediated at least in part by VEGFR-2. PMID- 15528048 TI - In vitro scavenging activity for reactive oxygen and nitrogen species by nonsteroidal anti-inflammatory indole, pyrrole, and oxazole derivative drugs. AB - This study was undertaken to evaluate the scavenging activity for reactive oxygen species (ROS) and reactive nitrogen species (RNS) by several nonsteroidal anti inflammatory drugs (NSAIDs), namely indole derivatives (indomethacin, acemetacin, etodolac), pyrrole derivatives (tolmetin and ketorolac), and an oxazole derivative (oxaprozin). The inhibition of prostaglandin synthesis constitutes the primary mechanism of the anti-inflammatory action of these drugs. Nevertheless, it has been suggested that the anti-inflammatory activity of NSAIDs may be also partly due to their ability to scavenge ROS and RNS and to inhibit the respiratory burst of neutrophils triggered by various activator agents. Thus, the scavenging activity of these NSAIDs was evaluated against an array of ROS (O(2)( ), HO, HOCl, and ROO) and RNS (NO and ONOO(-)) using noncellular in vitro systems. The results obtained demonstrated that tolmetin, ketorolac, and oxaprozin were not active against O(2)(-), while acemetacin, indomethacin, and etodolac exhibited concentration-dependent effects. Oxaprozin was also the least active scavenger for HO, among all the tested NSAIDs shown to be active. The scavenging effect for HOCl was not observed for any of the tested NSAIDs. The ROO was effectively scavenged by etodolac, with the other tested NSAIDs being much less active. NO and ONOO(-) were scavenged by all the tested NSAIDs. These effects may strongly contribute to the anti-inflammatory therapy benefits that may be attained with some of the studied NSAIDs. PMID- 15528049 TI - Plasma membrane gamma-glutamyltransferase activity facilitates the uptake of vitamin C in melanoma cells. AB - Adequate cellular transport of ascorbic acid (AA) and its oxidation product dehydroascorbate (DHA) is assured through specific carriers. It was shown that vitamin C is taken up as DHA by most cell types, including cancer cells, via the facilitative GLUT transporters. Thus, AA oxidation to DHA can be considered a mechanism favoring vitamin C uptake and intracellular accumulation. We have investigated whether such an AA-oxidizing action might be provided by plasma membrane gamma-glutamyltransferase (GGT), previously shown to function as an autocrine source of prooxidants. The process was studied using two distinct human metastatic melanoma clones. It was observed that the Me665/2/60 clone, expressing high levels of membrane GGT activity, was capable of effecting the oxidation of extracellular AA, accompanied by a marked increase of intracellular AA levels. The phenomenon was not observed with Me665/2/21 cells, possessing only traces of membrane GGT. On the other hand, AA oxidation and stimulation of cellular uptake were indeed observed after transfection of 2/21 cells with cDNA coding for GGT. The mechanism of GGT-mediated AA oxidation was investigated in acellular systems, including GGT and its substrate glutathione. The process was observed in the presence of redox-active chelated iron(II) and of transferrin or ferritin, i.e., two physiological iron sources. Thus, membrane GGT activity-often expressed at high levels in human malignancies-can oxidize extracellular AA and promote its uptake efficiently. PMID- 15528050 TI - The Burden of Stroke Scale (BOSS) provides valid and reliable score estimates of functioning and well-being in stroke survivors with and without communication disorders. AB - BACKGROUND AND OBJECTIVE: This study describes the conceptual foundation and psychometric properties of the Burden of Stroke Scale (BOSS), a patient-reported health status assessment designed to quantify the physical, cognitive, and psychological burden of stroke. METHODS: Qualitative research methods were used to develop a 112-item pilot version of the instrument. The pilot version was administered to healthy controls (n=251) and stroke survivors with (n=135) and without (n=146) communication disorders on a single occasion for the purposes of reducing the global item pool, describing the resulting scale properties, examining the dimensionality of the burden of stroke construct, and examining the known-groups validity of the instrument. RESULTS: Sixty-four items were retained, comprising 12 internally consistent and unidimensional scales. Principal components analysis revealed three second-order factors (Physical Activity Limitations, Cognitive Activity Limitations, and Psychological Distress) comprising the Burden of Stroke construct. Comparisons between groups revealed that stroke survivors reported greater activity limitations and psychological distress on all scales relative to controls, and that stroke survivors with communication disorders reported greater activity limitations on swallowing, communication, cognition, and social relations scales relative to non communicatively disordered stroke survivors. CONCLUSION: These findings support the internal consistency of the BOSS scales, the hypothesized dimensionality of the burden of stroke construct, and the known-groups validity of the instrument. PMID- 15528051 TI - Reliable change and minimum important difference (MID) proportions facilitated group responsiveness comparisons using individual threshold criteria. AB - OBJECTIVE: This study contrasted the use of responsiveness indices at the group level vs. individual patient level. STUDY DESIGN AND SETTING: We followed a cohort of 211 patients (50% male; mean age 47.5 years; SD 14) with musculoskeletal upper extremity problems for a total of 3 months. Outcome measures included the Disabilities of the Arm, Shoulder, and Hand (DASH) questionnaire, Shoulder Pain and Disability Index (SPADI), Patient-Rated Wrist Evaluation (PRWE), and the Medical Outcomes Study 12-Item Short-Form Health Survey (SF-12). We calculated confidence intervals on various group-level responsiveness statistics based on effect size and correlation with global change. The proportion of patients exceeding the minimum detectable change (or reliable change proportion) and minimum important difference (MID proportion) were included as indices applicable to the individual patient. RESULTS: For the DASH, effect size ranged from 1.06 to 1.67 for various patient subgroups, and the reliable change and MID proportions indicated that 50%-70% of individuals exhibited change based on individual change scores. Only the SRM and reliable change proportion indicated differences among the outcome measures used in this study. CONCLUSION: The reliable change and MID proportions have an intuitive interpretation and facilitate quantitative responsiveness comparisons among outcome measures based on individual patient criteria. PMID- 15528052 TI - A clinically meaningful difference was generated for a performance measure of recovery from hip fracture. AB - BACKGROUND AND OBJECTIVE: The Lower Extremity Gain Scale (LEGS) is a performance measure of tasks that are often impaired in hip fracture patients. This study was designed to determine a clinically meaningful difference in LEGS. METHODS: The population was 139 female patients (age >65 years) admitted to Baltimore hospitals. Recovery levels were estimated by fitting trajectory curves for the cohort for the 12 months post fracture. The clinically meaningful difference was evaluated using an anchor-based approach, examining the relationship between the LEGS recovery level and age. A second, distribution-based method used an effect size of .20. RESULTS: According to our model, a difference of 5 years in age corresponded to a difference of 1.6-3.6 points in LEGS scores. The standard deviation for LEGS at 12 months was 8.0; thus, Cohen's effect size of 0.2 would equate to a difference of 1.6 points. CONCLUSION: This suggests that a clinically meaningful difference in the LEGS scores for a population in this age range would be 2-3 points. PMID- 15528053 TI - Condition-specific Western Ontario McMaster Osteoarthritis Index was not superior to region-specific Lower Extremity Functional Scale at detecting change. AB - OBJECTIVE: We determined whether the sensitivity to change of the Western Ontario McMaster Osteoarthritis Index (WOMAC) physical function (PF) subscale, a condition-specific measure for persons with osteoarthritis of the hip or knee, was superior to a lower extremity region-specific measure, the Lower Extremity Functional Scale (LEFS), in persons with osteoarthritis of the hip or knee undergoing total joint arthroplasty. METHODS: The WOMAC and LEFS were administered to 102 patients with osteoarthritis preoperatively, within 16 days of surgery, and >20 days after the first postoperative assessment. These time points enabled the assessment of deterioration and improvement. Two timed performance measures (40 m walk test and the timed-up-and-go test) were also assessed. Change was quantified by the standardized response mean (SRM). RESULTS: WOMAC PF SRMs were not greater than the LEFS SRMs. Performance measures' times increased significantly over the deterioration interval and decreased over the improvement interval. The WOMAC PF and LEFS demonstrated significant improvement over the second assessment interval; only the LEFS showed significant deterioration over the first assessment interval. WOMAC PF scores seem to be strongly influenced by pain. CONCLUSION: The findings do not support the hypothesis that the WOMAC PF subscale is superior to the LEFS in detecting change. PMID- 15528054 TI - Disease adaptation may have decreased quality-of-life responsiveness in patients with chronic progressive neurological disorders. AB - OBJECTIVE: We hypothesized that disease adaptation could be measured in chronic progressive neurological disorders (CPND) through paired longitudinal comparisons of quality of life (QOL) and health status (HS) and of the mental health (MH) and physical health (PH) domains of QOL instruments. STUDY DESIGN AND SETTING: We identified 193 quantitative studies of QOL and HS in a systematic review of episodic (END) and chronic progressive (CPND) neurological disorders. Effect size or other responsiveness measures were analyzed in 31 studies that included paired longitudinal comparisons of QOL-HS, MH-PH, or both. Responsiveness means were compared using the paired-sample t-test or sign test. RESULTS: In 12 paired comparisons, QOL responsiveness was significantly lower than HS (P=.05, sign test). In 53 paired MH-PH effect-size comparisons, MH responsiveness was lower than PH (P=.02, t=2.48, paired sample). Significantly lower MH responsiveness was observed in 28 paired CPND comparisons (P < .01, t=3.86, paired sample) but not in 25 paired END comparisons (P=.50, t=0.68, paired sample). CONCLUSION: Lower responsiveness of QOL in CPND may be related to disease adaptation. Further prospective studies are needed to confirm our findings and to investigate the importance of disease adaptation in the evaluation of neurological disease and in health resource allocation. PMID- 15528055 TI - Common comorbidity scales were similar in their ability to predict health care costs and mortality. AB - OBJECTIVE: To compare the ability of commonly used measures of medical comorbidity (ambulatory care groups [ACGs], Charlson comorbidity index, chronic disease score, number of prescribed medications, and number of chronic diseases) to predict mortality and health care costs over 1 year. STUDY DESIGN AND SETTING: A prospective cohort study of community-dwelling older adults (n=3,496) attending a large primary care practice. RESULTS: For predicting health care charges, the number of medications had the highest predictive validity (R(2)=13.6%) after adjusting for demographics. ACGs (R(2)=16.4%) and the number of medications (15.0%) had the highest predictive validity for predicting ambulatory visits. ACGs and the Charlson comorbidity index (area under the receiver operator characteristic [ROC] curve=0.695-0.767) performed better than medication-based measures (area under the ROC curve=0.662-0.679) for predicting mortality. There is relatively little difference, however, in the predictive validity across these scales. CONCLUSION: In an outpatient setting, a simple count of medications may be the most efficient comorbidity measure for predicting utilization and health care charges over the ensuing year. In contrast, diagnosis-based measures have greater predictive validity for 1-year mortality. Current comorbidity measures, however, have only poor to moderate predictive validity for costs or mortality over 1 year. PMID- 15528056 TI - Statistical measures were proposed for identifying longitudinal patterns of change in quantitative health indicators. AB - OBJECTIVE: To propose statistical measures to identify different longitudinal patterns of change in quantitative health indicators. METHODS: The authors propose several simple measures to discriminate between stable-unstable, increasing-decreasing, linear-nonlinear, monotonic-nonmonotonic patterns of change. They then suggest using factor analysis to select the subset of nonredundant measures, and cluster analysis, based on the selected measures, to identify subgroups of patients with similar longitudinal trajectories. The proposed approach is illustrated using data on osteoarthritis disability from a longitudinal study undertaken in Toronto, Ontario, in 1996-2001. Disability was measured at four points in time for 835 patients, using the Western Ontario and McMaster Universities Osteoarthritis (WOMAC) index. RESULTS: The proposed measures allowed the detection of individual patients with specific patterns of change and identification of four different groups of patients with WOMAC scores that are (1) regularly increasing, (2) regularly decreasing, (3) stable over time, or (4) highly unstable, with abrupt changes or short-term fluctuations. CONCLUSION: The proposed approach may be used to (1) screen even large databases to identify particular patterns of change; or (2) summarize different patterns of change by classifying patients into groups with similar trajectories. Once identified, the groups can be investigated to determine whether there are differences in patient characteristics or outcomes. PMID- 15528057 TI - Mass mailing and telephone contact were effective in recruiting veterans into an antibiotic treatment randomized clinical trial. AB - OBJECTIVE: Achieving enrollment goals of randomized clinical trials (RCT) within budgets depends on the timely recruitment of sufficient numbers of participants. We report a comparison of recruitment methods and yields of previously deployed veterans into a large RCT. STUDY DESIGN AND SETTING: A retrospective survey concerning recruitment was administered to staff at 28 sites participating in the VA Cooperative Study #475, "Antibiotic Treatment of Gulf War Veterans' Illnesses" (GWVI). RESULTS: Twenty-one sites reported identifying 31,407 Gulf War Veterans (GWV). Of these, 13.7% were successfully contacted, 3.5% were enrolled, and 1.2% were randomized. Mass mailings and direct telephone calls to GWV identified from a GWV database accounted for 78% of the GWV contacted. The other 22% were contacted by using referrals from medical staff, veterans' groups, media advertisements, and other methods. Data collected prospectively at the Albany Stratton VAMC were similar to data collected retrospectively from other sites. CONCLUSION: These findings demonstrate that in previously deployed GWV with GWVI, 1.2% could be randomized. Although the use of all recruitment methods combined achieved the study recruitment goal, these data demonstrate that mass mailing and direct telephone contacts were effective recruitment methods. PMID- 15528058 TI - Misclassifying the indications for prostate-specific antigen testing may bias case-control studies of the efficacy of prostate cancer screening. AB - BACKGROUND AND OBJECTIVE: In the absence of data from randomized controlled trials, prostate cancer (CaP) screening recommendations may be based on observational studies that contrast exposure to screening between cases and controls. We evaluated the potential bias from mis-classifying indications for PSA testing in observational studies of CaP screening. METHODS: We randomly selected men undergoing PSA testing and obtained data on PSA results and prostate biopsies. Data were linked with a tumor registry to identify incident and prevalent cases of CaP. We abstracted medical records for 45 incident cases with CaP and 118 controls without, recording information on lower urinary tract symptoms (LUTS), constitutional symptoms, and digital rectal examination findings. PSA testing was classified as definitely, likely, or possibly screening, or not screening based on clinical history. RESULTS: Changing the definitions for PSA screening to variably exclude men with LUTS and enlarged prostates differentially lowered the frequency of screening. With more restrictive screening definitions, the odds ratio for screening decreased from 0.47 to 0.07. CONCLUSION: Accurately classifying PSA testing status is difficult because LUTS are common among men targeted for CaP screening. Failing to correctly classify PSA tests may bias study results. PMID- 15528059 TI - A flexible modeling approach to estimating the component effects of smoking behavior on lung cancer. AB - OBJECTIVE: Despite the established causal association between cigarette smoking and lung cancer, the relative contributions of age started, duration, years since quitting, and daily amount smoked have not been well characterized. We estimated the contribution of each of these aspects of smoking behavior. STUDY DESIGN AND SETTING: A case-control study was conducted in Montreal on the etiology of lung cancer. There were 640 cases and 938 control subjects for whom lifetime smoking histories were collected. We used generalized additive models, incorporating cubic smoothing splines to model nonlinear effects of various smoking variables. We adopted a multistep approach to deal with the multicollinearity among time related variables. RESULTS: The main findings are that (1) risk increases independently by daily amount and by duration; (2) among current smokers, lung cancer risk doubles for every 10 cigarettes per day up to 30 to 40 cigarettes per day and tails off thereafter; (3) among ex-smokers, the odds ratio decreases with increasing time since quitting, the rate of decrease being sharper among heavy smokers than among light smokers; and (4) absolute risks demonstrate the dramatic public health benefits of long-term smoking cessation. CONCLUSION: Our results reinforce some previous findings on this issue. PMID- 15528060 TI - Hematocrit was not validated as a surrogate end point for survival among epoetin treated hemodialysis patients. AB - OBJECTIVE: To evaluate the use of hematocrit as a surrogate end point for survival among end-stage renal disease (ESRD) patients treated with epoetin. STUDY DESIGN AND SETTING: Using United States Renal Data System (USRDS) data, we conducted an observational prospective study to analyze the relationships among epoetin dose, hematocrit, and survival for 31,301 facility-based hemodialysis patients incident to ESRD therapy in 1998. To address our objective, we used criteria developed by Prentice based on results from a Cox regression model. RESULTS: Results indicate that hematocrit is inversely associated with epoetin dose. For the same epoetin treatment-related achieved hematocrit levels, there were widely varying treatment-related survival outcomes, thereby challenging a central criterion required to empirically validate a surrogate end point. CONCLUSION: Our results support earlier clinical trial and epidemiological data suggesting that hematocrit may not be a valid surrogate for survival among the epoetin-treated renal failure population. We hypothesize that hematocrit may not be in the causal pathway or that epoetin may have important mechanisms of action apart from increasing hematocrit. Effective treatment for anemia may therefore not be simply a matter of increasing hematocrit. This study has potential implications for revising the existing treatment guidelines for anemia management and selecting an appropriate treatment regimen. PMID- 15528061 TI - Agreement between self-report questionnaires and medical record data was substantial for diabetes, hypertension, myocardial infarction and stroke but not for heart failure. AB - OBJECTIVES: Questionnaires are used to estimate disease burden. Agreement between questionnaire responses and a criterion standard is important for optimal disease prevalence estimates. We measured the agreement between self-reported disease and medical record diagnosis of disease. STUDY DESIGN AND SETTING: A total of 2,037 Olmsted County, Minnesota residents > or =45 years of age were randomly selected. Questionnaires asked if subjects had ever had heart failure, diabetes, hypertension, myocardial infarction (MI), or stroke. Medical records were abstracted. RESULTS: Self-report of disease showed >90% specificity for all these diseases, but sensitivity was low for heart failure (69%) and diabetes (66%). Agreement between self-report and medical record was substantial (kappa 0.71 0.80) for diabetes, hypertension, MI, and stroke but not for heart failure (kappa 0.46). Factors associated with high total agreement by multivariate analysis were age <65 years, female sex, education >12 years, and zero Charlson Index score (P < .05). CONCLUSION: Questionnaire data are of greatest value in life-threatening, acute-onset diseases (e.g., MI and stroke) and chronic disorders requiring ongoing management (e.g.,diabetes and hypertension). They are more accurate in young women and better-educated subjects. PMID- 15528062 TI - Clinicians required very high sensitivity of a bacteremia prediction rule. AB - BACKGROUND AND OBJECTIVE: Efforts to improve blood culture practice have focused on developing clinical prediction rules to identify patients at risk for bacteremia. However, no such models have been accepted into general clinical use. The goal of this study was to determine physicians' criteria for acceptability of a bacteremia clinical prediction rule. METHOD: We conducted a survey of all medical and surgical house officers as well as all infectious diseases physicians at the University of Pennsylvania to identify physician requirements for the sensitivity of a bacteremia clinical prediction rule. RESULTS: Of 225 eligible physicians, 149 (66.2%) completed the survey, including 110 house officers and 39 infectious disease physicians. The median (95% confidence interval) sensitivity of a bacteremia prediction rule required by respondents was 95% (95% confidence interval, 95.9%). Furthermore, 29 (19.5%) respondents required the sensitivity of a prediction rule to be at least 99%. The median required sensitivity was significantly higher for infectious diseases physicians than for house officers (98% and 95%, respectively) (P=.04). CONCLUSION: Our survey of house staff and infectious diseases physicians demonstrates that the sensitivity of any bacteremia prediction rule must be extremely high (i.e., 99% to 100%) to be widely accepted by practicing clinicians. Elucidation of physician criteria for acceptability of clinical predictive models will be invaluable in future efforts to develop such prediction rules. PMID- 15528064 TI - A graphical user interface for automatic image registration software designed for radiotherapy treatment planning. AB - Medical imaging forms a vital component of radiotherapy treatment planning and its evaluation. The integration of the useful data obtained from multiple imaging modalities for radiotherapy planning is achieved by image registration softwares. In radiotherapy planning systems, normally the computed tomography (CT) slices are kept as a standard upon which other modality images (magnetic resonance imaging [MRI], single photon emission computed tomography [SPECT], positron emission tomography [PET], etc.) are aligned--automatically or interactively. Following validation of successful registration, they are resampled and reformatted, as per the requirements. This paper defines the minimum requirements of automatic image registration software for 3-dimensional (3D) radiotherapy planning and describes the implementation of a suitable graphical user interface developed in Visual Basic (version 5). The automatic image registration (AIR) routines freely available from Dr. Roger P. Woods, UCLA, (USA) were used in this software. This software could be easily implemented and was easy to use for image processing suitable for radiotherapy planning systems. PMID- 15528063 TI - A time-to-prescription-refill measure of antiretroviral adherence predicted changes in viral load in HIV. AB - OBJECTIVE: The goal of this study was to determine the validity and utility of a pharmacy-based time-to-refill measure of antiretroviral therapy adherence. METHODS: We performed an observational cohort study of 110 HIV-infected subjects on a stable, highly active antiretroviral regimen for at least 3 months at a Veterans' Affairs Medical Center in Philadelphia, Pennsylvania. RESULTS: The viral load decreased by 0.12 log c/mL (95% confidence interval [CI] 0.01-0.23 log c/mL) for each 10% increase in pharmacy-based time-to-refill defined adherence as compared with 0.05 log c/mL (95% CI -0.14-0.25 log c/mL) for the self-reported adherence measure. Thus, only the refill-defined measure was statistically significantly associated with viral load change. When adherence was classified as good (> or = 85%) versus poor (<85%), both measures demonstrated a significant difference in outcome between groups. Yet, in individuals self-reporting 100% adherence, those classified as good adherers using the pharmacy-based measure had greater viral load reductions than poor adherers (2.4 log c/mL [interquartile range 1.4-3.4] vs. 1.5 log c/mL [interquartile range 0.8-2.4, P=.03). CONCLUSIONS: The pharmacy-based technique is a valid measure of antiretroviral therapy adherence. Because it provides clinically relevant information in subjects who self-report 100% adherence, it should be incorporated into clinical practice and adherence research. PMID- 15528065 TI - Application of intensity-modulated radiation therapy for pediatric malignancies. AB - Novel radiation therapy delivery techniques have moved very slowly in the field of pediatric oncology. Some collaborative groups allow new radiation therapy delivery techniques in their trials. In many instances, the option of using these techniques is not addressed. These newer techniques of radiation delivery have the potential to reduce the probability of the common late effects of radiation and at the same time, potentially improve upon control and survival. The purpose of this study is to show the feasibility of IMRT in pediatric patients. No treatment results or toxicities will be presented. Five patients with a variety of pediatric malignancies received intensity-modulated radiation therapy (IMRT) at our institution as part of their disease management. A rigid immobilization device was developed for each patient and a computed tomography (CT) simulation was performed in the treatment position. In 3 of the patients, magnetic resonance imaging (MRI) scans were coregistered with the planning CT to facilitate target and critical structure delineation. In all but 1 patient, coplanar beam arrangements were used in the IMRT planning process. All IMRT plans exhibited a high degree of conformality. Dose homogeneity inside the tumor and rapid dose falloff outside the target volume is characteristic of IMRT plans, which allows for improved normal tissue sparing. Dose distributions were obtained for all plans, as well as dose and volume relationship histograms, to evaluate the fitness of the plans. IMRT is a viable alternative to conventional treatment techniques for pediatric cancer patients. The improved dose distributions coupled with the ease of delivery of the IMRT fields make this technique very attractive, especially in view of the potential to increase local control and possibly improve on survival. PMID- 15528066 TI - Intensity-modulated radiation therapy reduces the dose to normal tissue in T2N0M0 squamous cell carcinoma of the glottic larynx. AB - The purpose of this paper was to compare intensity-modulated radiation therapy (IMRT) and conventional planning for T2N0M0 squamous cell carcinoma (SQCC) of the glottic larynx. Three patients with T2N0M0 SQCC are presented who were treated with IMRT. Conventional plans were also generated for comparison purposes. Isodose distributions and dose-volume histograms (DVHs) were generated for all the plans to evaluate the fitness of the plan as well as the differential benefit of IMRT vs. conventional treatment. The isodose distributions that were obtained by the IMRT plan are much more conformal to the planning target volume (PTV) and clearly show that less healthy tissue is subjected to a high-dose level, thus reducing toxicity. IMRT offers better comformality without compromising the PTV coverage and delivers less dose to normal tissues as compared to conventional radiation therapy in T2N0M0 SQCC of the glottic larynx. With an increase in conformality, it is expected to have an increase in the therapeutic ratio. PMID- 15528067 TI - Irradiating the groin nodes without breaking a leg: a comparison of techniques for groin node irradiation. AB - The purpose of this study was to determine the optimal technique for delivering postoperative radiotherapy for vulvar cancer and other tumors requiring treatment of the inguinal nodes. This project compared tumor coverage and normal tissue sparing for the 5 main radiotherapy techniques that are used to treat vulvar cancer. The intensity-modulated radiation therapy (IMRT) plan was undesirable because it resulted in an excessive dose to portions of the central pelvic structures. The photon thunderbird with skin match was unacceptable because it underdosed a portion of the groin region. The electron thunderbird was ideal for thin patients but was not applicable for most patients because of excessive dose to the skin and subcutaneous tissues. The photon through-and-through and the photon thunderbird with deep match were acceptable in most situations. In thin patients, where the depth of the inguinal vessels is less than 3 cm, the electron thunderbird is the technique of choice. In the average-sized patient, both the photon through-and-through and the photon thunderbird with deep match are reasonable options. The available literature suggests that the risk of femoral neck fracture or necrosis of the femoral head is approximately 11% at 5 years using the photon through-and-through technique. In our opinion, this is an acceptable price to pay for reliable node coverage, setup simplicity, and zero risk of overdose at field junctions. PMID- 15528068 TI - Forward-planned, multiple-segment, tangential fields with concomitant boost in the treatment of breast cancer. AB - We report on the utility of forward-planned, 3-dimensional (3D), multiple-segment tangential fields for radiation treatment of patients with breast cancer. The technique accurately targets breast tissue and the tumor bed and reduces dose inhomogeneity in the target. By decreasing excess dose to the skin and lung, a concomitant boost to the tumor bed can be delivered during the initial treatment, thereby decreasing the overall treatment time by one week. More than 120 breast cancer patients have been treated with this breast conservation technique in our clinic. For each patient, a 3D treatment plan based upon breast and tumor bed volumes delineated on computed tomography (CT) was developed. Segmented tangent fields were iteratively created to reduce "hot spots" produced by traditional tangents. The tumor bed received a concomitant boost with additional conformal photon beams. The final tumor bed boost was delivered either with conformal photon beams or conventional electron beams. All patients received 45 Gy to the breast target, plus an additional 5 Gy to the surgical excision site, bringing the total dose to 50 Gy to the boost target volume in 25 fractions. The final boost to the excision site brought the total target dose to 60 Gy. With minimum follow-up of 4 months and median follow-up of 11 months, all patients have excellent cosmetic results. There has been minimal breast edema and minimal skin changes. There have been no local relapses to date. Forward planning of multi segment fields is facilitated with 3D planning and multileaf collimation. The treatment technique offers improvement in target dose homogeneity and the ability to confidently concomitantly boost the excision site. The technique also offers the advantage for physics and therapy staff to develop familiarity with multiple segment fields, as a precursor to intensity-modulated radiation therapy (IMRT) techniques. PMID- 15528069 TI - A Monte Carlo brachytherapy study for dose distribution prediction in an inhomogeneous medium. AB - The purpose of this study was to present a theoretical analysis of how the presence of bone in interstitial brachytherapy affects dose rate distributions. This study was carried out using a Monte Carlo simulation of the dose distribution in homogeneous medium for 3 commonly used brachytherapy seeds. The 3 seeds investigated in this study are iridium-192 (192Ir) iodine-125 (125I), and palladium-103 (103Pd). The computer code was validated by comparing the specific dose rate (Lambda), the radial dose function g(r), and anisotropy function F(r,theta;) for all 3 seeds with the AAPM TG-43 dosimetry formalism and current literature. The 192Ir seed resulted in a dose rate of 1.115 +/- 0.001 cGy-hr(-1) U(-1), the 125I seed resulted in a dose rate of 0.965 +/- 0.006 cGy/h(-1)/U(-1), and the 103Pd seed resulted in a dose rate of 0.671 +/- 0.002 cGy/h(-1)/U(-1). The results for all 3 seeds are in good agreement with the AAPM TG-43 and current literature. The validated computer code was then applied to a simple inhomogeneous model to determine the effect bone has on dose distribution from an interstitial implant. The inhomogeneous model showed a decrease in dose rate of 2% for the 192Ir, an increase in dose rate of 84% for 125I, and an increase in dose rate of 83% for the 103Pd at the surface of the bone nearest to the source. PMID- 15528070 TI - A clinical application of an automated phantom-film QA procedure for validation of IMRT treatment planning and delivery. AB - To quantify the correlation between planned and delivered intensity-modulated radiation therapy (IMRT) dose distributions, IMRT plans for 37 prostate carcinoma patients were analyzed. IMRT treatment plans were converted into hybrid phantom plans using a commercially available treatment planning system and delivered to a specialized film phantom via a static-tomotherapy technique. The films were analyzed using a commercial film dosimetry system. Hybrid phantom axial dose maps and film images were normalized, registered to one another, and subtracted to calculate the overall relative dose difference throughout the entire film area on a pixel-by-pixel basis. The average percentage of pixels with dose-difference values greater than +/- 3% among analyzed hybrid patient plans was 8.6% +/- 3%. The average percentage of pixels with dose differences greater than +/- 5% was 1.7% +/- 1.0%. The number of pixels with more than +/- 10% dose differences was negligible. An initial subset of hybrid plans was used to develop a quantitative criterion to verify for positional accuracy based on dosimetric verification of intensity map of IMRT plans for prostate patients in our institution. Plans with less than 5% of the pixels outside the +/- 5% dose-difference range were accepted. This method could be implemented for other anatomical sites or treatment planning and delivery systems. PMID- 15528071 TI - Total skin high-dose-rate electron therapy dosimetry using TG-51. AB - An approach to dosimetry for total skin electron therapy (TSET) is discussed using the currently accepted TG-51 high-energy calibration protocol. The methodology incorporates water phantom data for absolute calibration and plastic phantom data for efficient reference dosimetry. The scheme is simplified to include the high-dose-rate mode conversion and provides support for its use, as it becomes more available on newer linear accelerators. Using a 6-field, modified Stanford technique, one may follow the process for accurate determination of absorbed dose. PMID- 15528073 TI - Signaling control of memory T cell generation and function. AB - Memory T cells exhibit low activation thresholds and mediate rapid effector responses when recalled by antigen; contrasting the higher activation threshold, slower responses and predominant IL-2 production by naive T cells. While the sequence of intracellular events coupling the T cell-receptor (TCR) to naive T cell activation is well characterized, biochemical control of memory T cell differentiation and function remains undefined. In this review, we will discuss recent developments in T cell-receptor signal transduction as they pertain to memory T cells, and will discuss how signal dampening may drive memory generation, and more efficient spatial organization of signaling molecules may promote rapid recall responses. PMID- 15528074 TI - CD4 T-cell memory. AB - CD4 T-cell memory is in some ways more enigmatic than CD8 T-cell memory. This is mostly due to the fact that CD4 T cells tend to expand far less in response to antigenic stimuli, thereby thwarting attempts at their detection during the course of an immune response. Nevertheless, there is a wide range of experimental models that have provided information regarding the survival and maintenance of CD4 memory cells, their functional capacities, their differentiation states and program of development following activation. The emerging picture is one of great versatility and functional heterogeneity as befits their central position within the immune system. PMID- 15528075 TI - CD8 T cell memory. AB - This review describes what is generally known about CD8 immune responses, and focus in the most recent advances in this domain. It also attempts to point to the areas where experimental evidence is contradictory or insufficient, and thus require further exploration and clarification. PMID- 15528076 TI - Kinetics and clonality of immunological memory in humans. AB - T-cell immunological memory consists largely of clones of proliferating lymphocytes maintained by antigenic stimulation and the survival and proliferative effects of cytokines. The duration of survival of memory clones in humans is determine by the Hayflick limit on the number of cell divisions, the rate of cycling of memory cells and factors that control erosion of telomeres, including mechanisms that control telomerase. PMID- 15528077 TI - Generation and maintenance of immunological memory. AB - The key feature of the adaptive immune response is its specificity and the ability to generate and maintain memory. Preexisting antibodies in the circulation and at the mucosa provide the first line of defense against re infection by extracellular as well as intracellular pathogens. Memory T cells are an important second line of defense against intracellular pathogens, and in particular against microbes that can cause chronic or latent infection. In this article we will review our current understanding of the generation and maintenance of B cell and T cell memory. PMID- 15528079 TI - Prediction error for free monetary reward in the human prefrontal cortex. AB - Making predictions about future rewards is an important ability for primates, and its neurophysiological mechanisms have been studied extensively. One important approach is to identify neural systems that process errors related to reward prediction (i.e., areas that register the occurrence of unpredicted rewards and the failure of expected rewards). In monkeys that have learned to predict appetitive rewards during reward-directed behaviors, dopamine neurons reliably signal both types of prediction error. The mechanisms in the human brain involved in processing prediction error for monetary rewards are not well understood. Furthermore, nothing is known of how such systems operate when rewards are not contingent on behavior. We used event-related fMRI to localize responses to both classes of prediction error. Subjects were able to predict a monetary reward or a nonreward on the basis of a prior visual cue. On occasional trials, cue-outcome contingencies were reversed (unpredicted rewards and failure of expected rewards). Subjects were not required to make decisions or actions. We compared each type of prediction error trial with its corresponding control trial in which the same prediction did not fail. Each type of prediction error evoked activity in a distinct frontotemporal circuit. Unexpected reward failure evoked activity in the temporal cortex and frontal pole (area 10). Unpredicted rewards evoked activity in the orbitofrontal cortex, the frontal pole, parahippocampal cortex, and cerebellum. Activity time-locked to prediction errors in frontotemporal circuits suggests that they are involved in encoding the associations between visual cues and monetary rewards in the human brain. PMID- 15528078 TI - CD8 memory T cells: cross-reactivity and heterologous immunity. AB - Virus-specific memory T cell populations demonstrate plasticity in antigen recognition and in their ability to accommodate new memory T cell populations. The degeneracy of T cell antigen recognition and the flexibility of diverse antigen-specific repertoires allow the host to respond to a multitude of pathogens while accommodating these numerous large memory pools in a finite immune system. These cross-reactive memory T cells can be employed in immune responses and mediate protective immunity, but they can also induce life threatening immunopathology or impede transplantation tolerance and graft survival. Here we discuss examples of altered viral pathogenesis occurring as a consequence of heterologous T cell immunity and propose models for the maintenance of a dynamic pool of memory cells. PMID- 15528080 TI - Dynamics of parietofrontal networks underlying visuospatial short-term memory encoding. AB - Brain imaging studies in TEP, functional magnetic resonance imaging (fMRI) and electroencephalography (EEG) have shown that visuospatial short-term memory tasks depend on dorsal parietofrontal networks. Knowing the spatiotemporal dynamics of this network would provide further understanding of the neural bases of the encoding process. We combined magnetoencephalography (MEG) with EEG and fMRI techniques to study this network in a task, in which participants had to judge the symmetry in position of two dots, presented either simultaneously ("immediate comparison") or successively ("memorization" of a first dot and "delayed comparison", after 3 s, with a second dot). With EEG, larger amplitude was observed in the parietocentral P3b component (350-500 ms) in the immediate and "delayed comparisons" than in "memorization" condition, where topography at this time was more anterior and right lateralized. MEG provided a more accurate localization and temporal variations of sources, revealing a strong M4 component at 450 ms in the "memorization" condition, with two sources localized in parietal and right premotor regions. These localizations are consistent with both fMRI foci and EEG cortical current source densities (CSD), but only MEG revealed the strong increase in premotor region at 450 ms related to "memorization". These combined results suggest that EEG P3B and MEG M4 components reflect two different dynamics in parietofrontal networks: the parietocentral P3b indexes a decision mechanism during the immediate and "delayed comparisons", whereas the MEG M4 component, with a larger right premotor source, reflects the encoding process in visuospatial short-term memory. PMID- 15528081 TI - Mapping IQ and gray matter density in healthy young people. AB - Magnetic resonance imaging (MRI) studies suggest that significant changes in gray matter density occur during adolescence because of brain maturation. It has also been reported that gray matter volume correlates with measures of intellectual ability. This study examined whether the relationship between general intellectual ability (IQ) and gray matter morphometry reflects differential involvement of particular cytoarchitectonic areas. We found positive correlations between IQ and gray matter density in the orbitofrontal cortex, cingulate gyrus, the cerebellum, and thalamus and negative correlations in the caudate nucleus. These findings suggest that general intellectual ability in healthy young people is related to specific brain regions known to be involved in the executive control of attention, working memory, and response selection. PMID- 15528082 TI - Hierarchical Bayesian estimation for MEG inverse problem. AB - Source current estimation from MEG measurement is an ill-posed problem that requires prior assumptions about brain activity and an efficient estimation algorithm. In this article, we propose a new hierarchical Bayesian method introducing a hierarchical prior that can effectively incorporate both structural and functional MRI data. In our method, the variance of the source current at each source location is considered an unknown parameter and estimated from the observed MEG data and prior information by using the Variational Bayesian method. The fMRI information can be imposed as prior information on the variance distribution rather than the variance itself so that it gives a soft constraint on the variance. A spatial smoothness constraint, that the neural activity within a few millimeter radius tends to be similar due to the neural connections, can also be implemented as a hierarchical prior. The proposed method provides a unified theory to deal with the following three situations: (1) MEG with no other data, (2) MEG with structural MRI data on cortical surfaces, and (3) MEG with both structural MRI and fMRI data. We investigated the performance of our method and conventional linear inverse methods under these three conditions. Simulation results indicate that our method has better accuracy and spatial resolution than the conventional linear inverse methods under all three conditions. It is also shown that accuracy of our method improves as MRI and fMRI information becomes available. Simulation results demonstrate that our method appropriately resolves the inverse problem even if fMRI data convey inaccurate information, while the Wiener filter method is seriously deteriorated by inaccurate fMRI information. PMID- 15528083 TI - A longitudinal fMRI study: in recovering and then in clinically stable sub cortical stroke patients. AB - The aim of this 1-year longitudinal fMRI study was to compare hand motor activation patterns between cerebrovascular paretic patients with a subcortical infarction and healthy elderly subjects and to evaluate the changes between the subacute phase and the chronic phase of recovery. We studied eight right-handed patients with pure motor hemiparesis due to a single ischemic infarct of the corticospinal tract. Each patient underwent a first fMRI (E1) 20 +/- 9 days after stroke, a second (E2) after 4 months and a third (E3) 12 months after stroke. During each fMRI session, the patients performed an active motor task consisting of audio-paced (1 Hz) finger flexion-extension of the paretic hand and underwent a passive motor task consisting of flexion-extension of the paretic hand performed by an examiner. Data were analyzed with SPM99 (random effect analyses). Patients had recovered at E2, were stable between E2 and E3, but still experienced a hand weakness. Displacement of activation maxima coordinates in patients compared to healthy subjects suggested an early reorganization within the SMA and a secondary reorganization within the ipsilesional S1M1 at E2. The main differences between patients and healthy subjects were (1) recruitment of the posterior part of the cingulate cortex and SMA, (2) a general hyperactivation (except in the deefferented primary motor cortex) and (3) an evolution in the S1M1 activation from an early (20 days after stroke) contralesional hyperactivation to a later (4 months after stroke) ipsilesional hyperactivation concomitant to recovery. Changes in activation were confirmed by the passive task that involved no effort and little attention. Despite clinical stability, changes in brain processing seemed to occur between E2 and E3 corresponding to a normalization of ipsilesional S1M1 activation, a decrease of bilateral cerebellar activation, and a progressive increase in SII-BA 40 activity suggesting evolving compensatory networks to sustain recovery. PMID- 15528084 TI - Is voice processing species-specific in human auditory cortex? An fMRI study. AB - Recent studies suggested a sensitivity of regions of the human superior temporal sulcus (STS) to the sound of the human voice. However, the question of the species specificity of this response is still open. Healthy adult volunteers were scanned in an event-related fMRI design to compare responses in the STS to human and animal vocalizations, as well as to control nonvocal sounds (e.g., musical instruments). Bilateral activation of anterior STS was observed for human vocalizations, when contrasted with both nonvocal sounds and animal vocalizations. Animal vocalizations, compared to nonvocal sounds, elicited a more restricted left STS activation, although this region responded even more strongly to human vocalizations. This study provides the first evidence suggesting a species specificity in STS responses to vocalizations in humans. PMID- 15528085 TI - Short-term modulation of regional excitability and blood flow in human motor cortex following rapid-rate transcranial magnetic stimulation. AB - Repetitive transcranial magnetic stimulation (rTMS) of the human primary motor cortex (M1) provides a means of inducing lasting changes in cortical excitability and synaptic activity. Here we combined rTMS with positron emission tomography of regional cerebral blood flow (rCBF) to examine how an rTMS-induced change in intracortical excitability of inhibitory circuits affects regional synaptic activity. In a first set of experiments, we gave 150 biphasic pulses of 5 Hz rTMS at 90% of active motor threshold to left M1 and used single- and paired-pulse TMS to assess the conditioning effects of rTMS on motor cortical excitability at rest. rTMS conditioning led to a selective decrease in short-latency intracortical inhibition (SICI) without affecting short-latency intracortical facilitation or corticospinal excitability. The decrease in SICI lasted for approximately 8 min. In a second experiment, we used the same rTMS protocol and measured changes in regional synaptic activity (as indexed by rCBF) during and for up to 14 min after the end of rTMS. Subthreshold 5 Hz rTMS induced a region specific increase in resting rCBF in the stimulated M1 lasting approximately 8 min. These results suggest that in the stimulated M1, temporary attenuation of SICI is paralleled by an increase in synaptic activity, consistent with reduced efficacy of intracortical GABA(A)-ergic synapses. The present findings demonstrate that short trains of low-intensity 5 Hz rTMS can be used to induce a transient change in function within a distinct cortical area. This opens up new possibilities for studying acute reorganization at the systems level in the intact human brain. PMID- 15528086 TI - Comparison of standard and optimized voxel-based morphometry for analysis of brain changes associated with temporal lobe epilepsy. AB - We compared statistical parametric maps (SPMs) of group-wise regional gray matter differences between temporal lobe epilepsy (TLE) patients with unilateral hippocampal atrophy (HA) determined by manual volumetric analysis relative to a healthy control population using standard and optimized voxel-based morphometry (VBM). We also investigated the impact of customized neuroanatomical templates on SPMs. Standard and optimized VBM analyses of gray matter concentration (GMC) and gray matter volume (GMV) correctly identified HA, regardless of the template used for normalization. The distribution of hippocampal and extrahippocampal abnormalities differed according to the technique (standard v optimized; GMC v GMV), but was not dependent on template type (default v customized) within each technique. In particular, hippocampal GMC reduction was confined to subregions of hippocampus, whereas GMV reduction was observed in the hippocampal head, body, and tail. Unlike standard and optimized GMC reduction, symmetrical GMV reduction was observed in bilateral thalamus, lenticular nuclei, cerebellum, and ipsilateral entorhinal cortex, perirhinal cortex, and fusiform gyrus in both left and right HA patients. These results show that group-wise SPMs of GMC (i.e., regional distribution of gray matter) and GMV (i.e., volume per se) reduction can identify focal atrophy that has been quantified with manual region of interest techniques, although effects are attenuated in analyses of GMC. Unlike SPMs of GMC, analyses of GMV revealed similar extrahippocampal abnormalities as previous region-of-interest volumetric and histopathological studies of intractable TLE. We suggest that in studies of neurological disorders, optimized VBM analyses of GMV may reveal subtle neuroanatomical changes that are not identified in analyses of GMC. PMID- 15528087 TI - Hippocampal gray matter reduction associates with memory deficits in adolescents with history of prematurity. AB - Using optimized voxel-based morphometry (VBM), we compared the relationship between hippocampal and thalamic gray matter loss and memory impairment in 22 adolescents with history of prematurity (HP) and 22 normal controls. We observed significant differences between groups in verbal learning and verbal recognition, but not in visual memory. VBM analysis showed significant left hippocampal and bilateral thalamic reductions in HP subjects. Using stereological methods, we also observed a reduction in hippocampal volume, with left posterior predominance. We found correlations between left hippocampal gray matter reductions (assessed by VBM) and verbal memory (learning and percentage of memory loss) in the premature group. The stereological analysis showed a correlation between verbal learning and the left posterior hippocampus. Our results suggest that left hippocampal tissue loss may be responsible for memory impairment and is probably related to the learning disabilities that HP subjects present during schooling. PMID- 15528088 TI - Implicit and explicit processing of kanji and kana words and non-words studied with fMRI. AB - Using functional magnetic resonance imaging (fMRI), we investigated the implicit language processing of kanji and kana words (i.e., hiragana transcriptions of normally written kanji words) and non-words. Twelve right-handed native Japanese speakers performed size judgments for character stimuli (implicit language task for linguistic stimuli), size judgments for scrambled-character stimuli (implicit language task for non-linguistic stimuli), and lexical decisions (explicit language task). The size judgments for scrambled-kanji stimuli and scrambled-kana stimuli produced activations on the bilateral lingual gyri (BA 18), the bilateral occipitotemporal regions (BA 19/37), and the bilateral superior and inferior parietal cortices (BA 7/40). Interestingly, besides these areas, activations of the left inferior frontal region (Broca's area, BA 44/45) and the left posterior inferior temporal cortex (PITC, BA 37), which have been considered as language areas, were additionally activated during size judgment for kanji character stimuli. Size judgment for kana character stimuli also activated Broca's area, the left PITC, and the left supramarginal gyrus (SMG, BA 40). The activations of these language areas were replicated in the lexical decisions for both kanji and kana. These findings suggest that language processing of both kanji and kana scripts is obligatory to literate Japanese subjects. Moreover, comparison between the scrambled kanji and the scrambled kana showed no activation in the language areas, while greater activation in the bilateral fusiform gyri (left-side predominant) was found in kanji vs. kana comparison during the size judgment and the lexical decision. Kana minus kanji activated the left SMG during the size judgment, and Broca's area and the left middle/superior temporal junction during the lexical decision. These results probably reflect that in implicit or explicit reading of kanji words and kana words (i.e., hiragana transcriptions of kanji words), although using largely overlapping cortical regions, there are still some differences. Kanji reading may involve more heavily visual orthographic retrieval and lexical-semantic system through the ventral route, while kana transcriptions of kanji words require phonological recoding to gain semantic access through the dorsal route. PMID- 15528089 TI - The coupled dipole model: an integrated model for multiple MEG/EEG data sets. AB - Often MEG/EEG is measured in a few slightly different conditions to investigate the functionality of the human brain. This kind of data sets show similarities, though are different for each condition. When solving the inverse problem (IP), performing the source localization, one encounters the problem that this IP is ill-posed: constraints are necessary to solve and stabilize the solution to the IP. Moreover, a substantial amount of data is needed to avoid a signal to noise ratio (SNR) that is too poor for source localizations. In the case of similar conditions, this common information can be exploited by analyzing the data sets simultaneously. The here proposed coupled dipole model (CDM) provides an integrated method in which these similarities between conditions are used to solve and stabilize the inverse problem. The coupled dipole model is applicable when data sets contain common sources or common source time functions. The coupled dipole model uses a set of common sources and a set of common source time functions (STFs) to model all conditions in one single model. The data of each condition are mathematically described as a linear combination of these common spatial and common temporal components. This linear combination is specified in a coupling matrix for each data set. The coupled dipole model was applied in two simulation studies and in one experimental study. The simulations show that the errors in the estimated spatial and temporal parameters decrease compared to the standard separate analyses. A decrease in position error of a factor of 10 was shown for the localization of two nearby sources. In the experimental application, the coupled dipole model was shown to be necessary to obtain a plausible solution in at least 3 of 15 conditions investigated. Moreover, using the CDM, a direct comparison between parameters in different conditions is possible, whereas in separate models, the scaling of the amplitude parameters varies in general from data set to data set. PMID- 15528090 TI - Distinct representations for facial identity and changeable aspects of faces in the human temporal lobe. AB - The neural system underlying face perception must represent the unchanging features of a face that specify identity, as well as the changeable aspects of a face that facilitate social communication. However, the way information about faces is represented in the brain remains controversial. In this study, we used fMR adaptation (the reduction in fMRI activity that follows the repeated presentation of identical images) to ask how different face- and object-selective regions of visual cortex contribute to specific aspects of face perception. We report that activity in the face-selective region of the fusiform gyrus (FG) was reduced following repeated presentations of the same face. Adaptation in this area was not sensitive to changes in image size, but was sensitive to changes in viewpoint. In contrast, face-selective regions in the superior temporal lobe failed to adapt to identical presentations of the same face, but showed an increased response when the same face was shown from different viewpoints and with different expressions. These results reveal a largely size-invariant neural representation in the inferior temporal lobe that could be involved in the recognition of facial identity, and a separate face-selective region in the superior temporal lobe that could be used to detect changeable aspects of faces. The absence of fMR-adaptation in object-selective regions of visual cortex challenges the idea that a more distributed network of areas is used to represent information about faces. PMID- 15528091 TI - Abnormal brain activity in prefrontal brain regions in abstinent marijuana users. AB - We used PET (15)O and a modified version of the Stroop task to determine if 25 day abstinent heavy marijuana (MJ) users have persistent deficits in executive cognitive functioning (ECF) and brain activity. Performance on a modified version of the Stroop task and brain activity was compared between 25-day abstinent, heavy marijuana users (n = 11), and a matched comparison group (n = 11). The 25 day abstinent marijuana users showed no deficits in performance on the modified version of the Stroop task when compared to the comparison group. Despite the lack of performance differences, the marijuana users showed hypoactivity in the left perigenual anterior cingulate cortex (ACC) and the left lateral prefrontal cortex (LPFC) and hyperactivity in the hippocampus bilaterally, when compared to the comparison group. These results suggest that marijuana users display persistent metabolic alterations in brain regions responsible for ECF. It may be that marijuana users recruit an alternative neural network as a compensatory mechanism during performance on a modified version of the Stroop task. These differences in brain activity may be a common denominator in the evolution of maladaptive behaviors such as substance abuse and other neuropsychiatric disorders. PMID- 15528092 TI - When less means more: deactivations during encoding that predict subsequent memory. AB - In event-related functional MRI (fMRI) studies, greater activity for items that are subsequently remembered (R-items) than for items that are subsequently forgotten (F-items), or Dm effect (Difference in memory), has been attributed to successful encoding operations. In contrast, regions showing a reverse DM effect (revDM = F-items > R-items) have been linked to detrimental processes leading to forgetting. Yet, revDMs may reflect not only activations for F-items (aFs) but also deactivations for R-items (dRs), and the latter alternative is more likely to reflect beneficial rather than detrimental encoding processes. To investigate this issue, we used a paradigm that included a fixation baseline and could distinguish between the two types of revDMs (aF vs. dR). Participants were scanned while encoding semantic associations between words or perceptual associations between words and fonts, and their memory was measured with associative recognition tests. For both semantic and perceptual encoding, dR effects were found in dorsolateral prefrontal, temporoparietal, and posterior midline regions. In contrast with a prior study that attributed revDMs in these regions to detrimental processes, the present results suggest that these effects reflect beneficial processes, that is, the efficient reallocation of neurocognitive resources. At the same time, aF effects were found in other regions, such as the insula, and these are more consistent with an interpretation in terms of detrimental processes. Whereas most fMRI studies of encoding have focused on activation increases, the present study indicates that activation decreases are also critical for successful learning of new information. PMID- 15528093 TI - Regional patterns of brain metabolites in AIDS dementia complex. AB - The relationship of the cellular changes in the HIV-infected brain to the onset and progression of AIDS dementia complex (ADC) remains uncertain. We undertook an in vivo proton magnetic resonance spectroscopy (MRS) study and used factor analysis to identify specific cellular and regional brain changes that may serve as metabolic markers of ADC. The ratio of N-acetyl aspartate (NAA), choline (Cho), and myoinositol (MI) over creatine (Cr), markers of neuronal and glial cell metabolism, were measured in the basal ganglia, centrum semiovale, and parietal cortex from 100 subjects with and without ADC. Three metabolic patterns were identified, which we termed "inflammatory" (mainly MI/Cr elevations in all three regions plus Cho/Cr increases in the centrum semiovale and parietal cortex), "basal ganglia" (mostly NAA/Cr and Cho/Cr elevations in the basal ganglia), and "neuronal" (primarily NAA/Cr reductions in the centrum semiovale and the parietal cortex). Logistic regression analysis revealed that, adjusted for age, basal ganglia and neuronal pattern scores were strongly associated with ADC but inflammatory levels were not. We conclude that by using factor analysis, we are able to combine multiple metabolites across brain regions in a biologically plausible manner and construct a predictive model of ADC adjusting for relevant factors such as age. PMID- 15528094 TI - A general linear model for MEG beamformer imaging. AB - A new general linear model (GLM) beamformer method is described for processing magnetoencephalography (MEG) data. A standard nonlinear beamformer is used to determine the time course of neuronal activation for each point in a predefined source space. A Hilbert transform gives the envelope of oscillatory activity at each location in any chosen frequency band (not necessary in the case of sustained (DC) fields), enabling the general linear model to be applied and a volumetric T statistic image to be determined. The new method is illustrated by a two-source simulation (sustained field and 20 Hz) and is shown to provide accurate localization. The method is also shown to locate accurately the increasing and decreasing gamma activities to the temporal and frontal lobes, respectively, in the case of a scintillating scotoma. The new method brings the advantages of the general linear model to the analysis of MEG data and should prove useful for the localization of changing patterns of activity across all frequency ranges including DC (sustained fields). PMID- 15528095 TI - An fMRI study of the cerebral macro network involved in 'cue invariant' form perception and how it is influenced by stimulus complexity. AB - We investigated the influence of stimulus complexity on the macro network of visual areas involved in 'cue invariant' form perception. Functional MRI imaging on 14 healthy, adult volunteers was performed during a two alternative forced choice (2-AFC) form discrimination task. The functional load imposed onto the visual system was varied by using simple and complex shapes. The figures were defined using a luminance, a chromatic or a motion contrast cue. The three cues activated the same visual areas in the ventral pathway, including area 'LO'. Activation of visual area 'V3v' but not area 'KO' in the dorsal pathway was observed to the motion contrast cue. The simple shapes induced a larger BOLD response in BA18 than the complex shapes, reflecting the selectivity of this region for the features in the stimuli such as edges and vertices. The brain regions yielding a larger BOLD signal to the complex shapes were areas know to be selective to the orthographic content of our complex stimuli. The processing requirement was assessed by comparing the subjects' reaction time. We found no significant difference in the reaction times to the simple and complex shapes. The reaction times to the luminance contrast cue and the chromatic contrast cue were identical but that to the motion contrast cue were 200 ms longer. This finding concurs with neurophysiological studies, reporting a longer onset latency for motion contrast stimuli. It further lends support to the idea that the motion contrast cue requires auxiliary processing by the visual areas of the dorsal pathway before entry into the ventral pathway. PMID- 15528096 TI - The application of statistical parametric mapping to 123I-FP-CIT SPECT in dementia with Lewy bodies, Alzheimer's disease and Parkinson's disease. AB - Dopaminergic loss can be visualised using (123)I-FP-CIT single photon emission computed tomography (SPECT) in several disorders including Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Most previous SPECT studies have adopted region of interest (ROI) methods for analysis, which are subjective and operator-dependent. The purpose of this study was to investigate differences in striatal binding of (123)I-FP-CIT SPECT using the automated technique of statistical parametric mapping (SPM99) in subjects with DLB, Alzheimer's disease (AD), PD and healthy age-matched controls. This involved spatial normalisation of each subject's image to a customised template, followed by smoothing and intensity normalisation of each image to its corresponding mean occipital count per voxel. Group differences were assessed using a two-sample t test. Applying a height threshold of P or = 5 x 10(6)/kg and CD3 < 1 x 10(5)/kg recipient body weight), (2) toxicity, (3) survival to day +100. ELIGIBILITY: All hematological malignancies and ages; accrual to end after 20 transplants of patients with AML in remission and age less than 55 years. METHODS: Preparation: Modified Perugia regimen, chemotherapy alone; melphalan 140 mg/m2 day -9, thiotepa 10 mg/kg day -7, fludarabine 40 mg/m2 days -7 to -3, and ATG (Thymoglobulin, Sangstat) days -6 to -2 (total 10.5 mg/kg). Infection prophylaxis: Ganciclovir (GC) 5 mg/kg days 5-20 then x5/week until day +100 then x3/week until 210 (subjects 1-3), foscarnet (FC) 90 mg/kg days 4-21 then short course pre-emptive GC or FC (subjects 4-11); fluconazole; cotrimoxazole. Donors: G-CSF 16 microg/kg daily x5 until second pheresis day. T-cell depletion: CliniMACS (MiltenyiBiotec). RESULTS: Eleven patients with AML have been transplanted from four centers, eight female, three male, median age 34 (range 19 60). Disease status, first CR 1/11, second CR 4/11, third CR1/11, relapse 5/11. Graft CD34+ > or = 5 x 10(6)/kg was achieved in all cases, median 13.72 x 10(6)/kg (Q1, Q3: 8.26, 17.72; min 5.59, max 22.22), and CD3+ was < 1 x 10(5)/kg in all cases, median of 0.49 x 10(4)/kg (Q1, Q3: 0.30, 2.20; min 0.22, max 4.10). Ten of the 11 patients have died, median survival 103.5 days (Q1, Q3: 61.0, 151.0; min 0, max 290.0). Survival to day +100 6/11 (55%). Four patients died of leukemic relapse, six of infection. Of six patients dying of infection, CMV was a definite cause in four. Of four dying with relapse, CMV was significant in one. Engraftment was assessed in 10 patients who survived >0 days. Granulocyte engraftment (> 0.5 x 10(9)/l) was achieved in all patients, median 11.5 days (Q1, Q3: 10, 17; min 8, max 70). Platelet engraftment (> 20 x 10(9)/l) was achieved in 8 of 10 patients, median 15 days (Q1, Q3: 9, 16; min 9, max 97). The two platelet non-engrafters died on days +45 and +61. Toxicity was low, with one toxic death (day 0), and the Bearman organ toxicity gradings were < or = grade 2 in all other patients. There were no instances of graft-vs.-host disease or graft rejection. CONCLUSIONS: The problems of graft-vs.-host disease and graft rejection have been removed as barriers to haploidentical transplantation but the slow immune reconstitution limits its general application. Late referrals contribute to a high relapse rate and have delayed an optimal evaluation of the procedure. PMID- 15528136 TI - Human bone marrow CD34+ progenitor cells mature to T cells on OP9-DL1 stromal cell line without thymus microenvironment. AB - In this paper, we confirm data reported by the group of Zuniga-Pflucker that human cord blood CD34(+)38(-)Lin- progenitor cells when co-cultured with the murine stromal cell line OP9-DL engineered to express the Notch ligand delta-like 1 mature into T lymphocytes with a phenotypic progression as the one seen in thymus. We show that this is also the case for human T cells starting from CD34(+) adolescent bone marrow cells. These findings offer the theoretical possibility to generate ex vivo human T cells and administer them in vivo in patients to overcome their immune deficient window period after transplantation. However, the practical and theoretical problems that this new technology has to overcome before this technique can be applied in clinic are still enormous and discussed. PMID- 15528137 TI - Facilitating T-cell immune reconstitution after haploidentical transplantation in adults. AB - Delayed reconstitution of cellular immunity following T-cell-depleted, CD34 enriched, allogeneic hematopoietic progenitor cell transplantation (HPCT) is the major cause of morbidity and mortality following haploidentical transplantation in adults. This is illustrated in our recent study of 28 high-risk adult patients (median age 31) who were treated with conditioning regimens containing antithymocyte globulin (ATG) before T-cell-depleted, CD34-enriched allogeneic HPCT. Overall mortality was 93% (26/28 patients) with a median survival of 4 months posttransplant. Poor cellular immune reconstitution contributed to death of 21/28 patients, with eight deaths due to opportunistic infections and seven deaths due to relapse. While recovery of normal numbers of circulating NK cells and B-cells occurred within the first 1-2 months posttransplant, recovery of normal numbers of blood T-cells was suppressed for more than 1 year. The mean half-life of active ATG levels in serum was 6 days; rapid clearance suggested that residual ATG did not contribute to the delay of posttransplant T-cell reconstitution. Rapid T-cell reconstitution was seen only in younger patients, indicating that poor thymic function and the absence of T-cells in the graft are the major causes of delayed recovery of cellular immunity. Improved cellular immunity after T-cell-depleted haploidentical HPCT will thus require novel strategies to adoptively transfer antigen specific donor T-cells without inducing lethal graft-versus-host disease (GvHD). This problem has been addressed in a preclinical murine model of MHC-mismatched bone marrow transplantation. Donor T cells treated ex vivo with fludarabine or a UVA light-activated psoralen compound (amotosalen) have a markedly reduced ability to induce GvHD, yet the treated T cells confer protection against murine cytomegalovirus and an infused leukemic cell line. Polyclonal donor T-cells reconstituted the blood and lymphoid compartments posttransplant and expanded in vivo. Derivatives of ex-vivo-treated donor T-cells retained the ability to produce cytokines and proliferate in response to antigen challenge. The mechanism of reduced GvHD potential of ex-vivo treated T-cells appears to be selection of a subset of memory donor T-cells that do not initially home to secondary lymphoid organs and have reduced capacity for producing inflammation in the immediate posttransplant period. Direct selection of the memory subset by high-speed FACS confirmed the improved therapeutic index in the murine model system. Preclinical data indicate the feasibility of treating human T-cells with fludarabine, psoralen, or direct selection based upon the memory phenotype to efficiently produce a population of polyclonal donor T-cells with reduced GvHD activity. A planned clinical phase 1 trial of adoptive therapy utilizing ex vivo psoralen-treated donor T-cells in recipients of T-cell-depleted haploidentical HPCT is presented. PMID- 15528138 TI - Depletion of alloantigen-primed lymphocytes overcomes resistance to allogeneic bone marrow in mildly conditioned recipients. AB - OBJECTIVE: Successful implantation of allogeneic bone marrow (BM) cells after nonmyeloablative conditioning would allow to compensate for the inadequate supply of compatible grafts and to reduce mortality of graft-vs.-host disease (GVHD). Recently, we proposed to facilitate engraftment of mismatched BM by conditioning for alloantigen-primed lymphocyte depletion (APLD) with cyclophosphamide (CY). Here we summarize the experimental results obtained by this approach. MATERIALS AND METHODS: Naive or mildly irradiated BALB/c mice were primed with C57BL/6 BM cells (day 0), treated with CY (day 1) to deplete alloantigen-primed lymphocytes, and given a second C57BL/6 BM transplant (day 2) for engraftment. Recipients were repeatedly tested for chimerism in the blood and followed for GVHD and survival. The protocol was also tested for inducing tolerance to donor tissue and organ allografts, and for treatment of leukemia, breast cancer, and autoimmune diabetes in NOD mice. RESULTS: APLD by 200 mg/kg CY provided engraftment of allogeneic BM from the same donor in 100% mildly irradiated recipients. Eighty percent chimeras remained GVHD-free more 200 days. All chimeras accepted permanently donor skin grafts and donor hematopoietic stromal progenitors. Allogeneic BM transplantation (BMT) after APLD had a strong therapeutic potential in BALB/c mice harboring malignant cells and in autoimmune NOD recipients. Tolerance-inducing CY dose could be reduced to 100 mg/kg. Conditioning for APLD resulted in engraftment of allogeneic BM after a significantly lower radiation dose than treatment with radiation and CY alone. CONCLUSION: Our results demonstrate that conditioning for APLD has a definite advantage over general immunosuppression with CY and radiation therapy. PMID- 15528140 TI - Growth factors and DLI in adult haploidentical transplant: a three-step pilot study towards patient and disease status adjusted management. AB - Haploidentical transplant is now established as a procedure of choice for patients who lack a compatible donor. However, they are still referred too late, heavily pretreated, at very advanced stages. We initiated a three-step phase I study trying improve transplant-related mortality, relapse rate, and immunity: G CSF + DLI, GM-CSF + DLI, patient- and disease-adapted strategy. Thirty-three consecutive leukemia patients, aged 18-55, were investigated (20 very poor risk, 11 poor risk, and 2 better risk). GvH type NK alloreactivity was chosen when possible (18/33) and balanced across the three groups. In the first nine patients, G-CSF was used and escalated prophylactic DLI started at month 1. Thus, G-CSF and 1-3 DLI (10(4) CD3/kg) is safe. It results in faster CD4 recovery and a low rate of infections. However, it was insufficient to induce a GVL effect. In the next 12 patients, GM-CSF was used plus 1 DLI (10(4) CD3/kg) at day 30 unless aGVHD (3 patients). The comparison between the two first groups can be summarized as follows: G-CSF + DLI: TRM at day 100: 0, RR: 6/9, severe aGVHD: 0. GM-CSF + 1 DLI group: RR: 1/12, TRM at day 100: 3, aGVHD > 1: 9/12, price to pay: GVHD resulting in five deaths in total. Step 3 (13 patients) consists of a patient adapted strategy: no more aspecific DLI (selected anti-CMV and aspergillus DLI planned in all patients); in myeloid disorders with NK alloreactivity: no GF. In the other cases, GM-CSF (at a reduced total dose of 500 mug) is given the follow up of these 13 patients, although promising is currently short (median 5 months). Overall, TRM at day 100 is 3/29, reflecting the good tolerance of the conditioning in a heavily pretreated population (median age: 43). NRR mortality (8/26) at 1 year is greater in the GM-CSF + DLI group, reflecting the impact of severe aGVHD. We conclude that the third strategy might improve the outcome without exposing patients to unnecessary severe GVHD. PMID- 15528139 TI - Prospects for dendritic cell vaccination against fungal infections in hematopoietic transplantation. AB - Dendritic cells (DCs) are uniquely able to initiate and control the immune response to fungi. DCs function at three levels in the manipulation of the immune response to these pathogens. First, they mount an immediate or innate response to them, for example, by producing inflammatory mediators upon capture and phagocytosis; second, through these preceding innate functions, they decode the fungus-associated information and translate it in qualitatively different Th responses, and third, they are key in containing and dampening inflammatory responses by tolerization through the induction of regulatory T cells (Treg). DCs sense fungi in a morphotype-specific manner, through the engagement of distinct recognition receptors ultimately affecting cytokine production and costimulation. Both myeloid and plasmacytoid murine and human DCs phagocytose fungi and undergo functional maturation in response to them. However, their activation program for cytokine production was different, being IL-12 mainly produced by myeloid DCs and IL-12, IL-10 and IFN-alpha mainly produced by plasmacytoid DCs. This resulted in a distinct ability for T cell priming, being Th1, Th2, and Treg differently activated by the different DC subsets. The ability of fungus-pulsed DCs to prime for Th1 and Th2 cell activation upon adoptive transfer in vivo correlated with the occurrence of resistance and susceptibility to the infections, respectively. Antifungal protective immunity was also induced upon adoptive transfer of DCs transfected with fungal RNA. The efficacy was restricted to DCs transfected with RNA from yeasts or conidia but not with RNA from fungal hyphae. The effect was fungus-specific, as no cross-protection was observed upon adoptive transfer of DCs pulsed with either fungal species. The infusion of fungus-pulsed or RNA transfected DCs accelerated the recovery of functional antifungal Th1 responses in mice with allogeneic hematopoietic stem cell transplantation (HSCT) and affected the outcome of the infections. As the ability of phagocytose fungi was defective in peripheral DCs from patients with HSCT, soon after the transplant, our findings suggest that the adoptive transfer of DCs may restore immunocompetence in HSCT by contributing to the educational program of T cells. Thus, the remarkable functional plasticity of DCs in response to fungi can be exploited for the deliberate targeting of cells and pathways of cell-mediated immunity in response to fungal vaccines. PMID- 15528141 TI - IL-2 activated NK cell immunotherapy of three children after haploidentical stem cell transplantation. AB - Natural killer (NK) cells are thought to be of benefit in HLA-mismatched hematopoietic transplantation (H-SCT). Therefore, we developed a protocol for clinical-use expansion of highly enriched and IL-2-stimulated NK cells. Purification of unstimulated leukaphereses by a two-step T cell depletion with a final CD56 enrichment procedure leads to a mean purity of 95% CD56(+)CD3- NK cells with a four- to five-log depletion of T cells. So far, three pediatric patients with multiply relapsed acute lymphoblastic leukemia (ALL) or acute myelogenous leukemia (AML) were treated with repeated transfusions post-H-SCT. Directed killer immunoglobulin-like receptor (KIR) mismatches were demonstrated in all three cases. Although all patients showed blast persistence at the time of transplant, they reached complete remission and complete donor chimerism within 1 month post-H-SCT. NK cell therapy was tolerated well without graft-versus-host disease (GvHD) induction or other adverse events. The AML patient died of early relapse on day +80, while the ALL patients died of thrombotic-thrombocytopenic purpura and atypical viral pneumonia on days +45 and +152, respectively. This initial trial showed the feasibility of good manufacturing practice (GMP) compliant NK cell isolation and expansion for clinical applications. We now launch a clinical phase I trial with activated NK cells post-H-SCT. PMID- 15528142 TI - Mesenchymal cells: a vehicle for gene therapy. AB - T cell-depleted allogeneic stem cell transplantation is associated with delayed immunological reconstitution. Bone marrow stroma and interleukin 7 (IL-7) regulate homeostasis of T lymphocytes. We engineered human stromal cells with a retroviral vector containing the IL-7 gene and studied in vitro effects on T cells. Human stromal cells were successfully transduced and generated a layer that was morphologically and phenotypically normal. IL-7-engineered stromal cells conserve the biological properties of unmanipulated stromal cells. Through their production of IL-7, they enhance survival and homeostatic proliferation of naive T cells. Because of this cytokine production, they might be an ideal vehicle for gene therapy aimed at supporting lymphopoiesis in the T cell-deficient host. PMID- 15528143 TI - Cord blood transplantation for children with acute leukaemia: a Eurocord registry analysis. AB - OBJECTIVES: Report results of unrelated cord blood transplants collected by Eurocord Registry in children with acute leukemia. RESULTS: Children with AML: 95 children were analyzed. The two year leukemia free survival was 42% in patients transplanted in first remission, 50% in second remission and 21% in children not in remission. Children with poor prognostic cytogenetic features had the same survival compared to other patients. Children with ALL: 195 patients with ALL were analyzed. The two year leukemia free survival was 36% in patients transplanted in remission and 15% in patients transplanted in relapse. Results of unrelated cord blood transplants compared to unrelated bone marrow transplants in children with acute leukemia: 416 children with acute leukemia received a HLA matched unrelated bone marrow transplant and were compared to 99 children transplanted with an unrelated HLA mismatched cord blood. The long term outcome between these groups were comparable with delayed engraftment in cord blood transplant, more relapse in T cell depleted bone marrow transplant and more GVH in the unmanipulated bone marrow transplant resulting in similar 5 years leukemia free survival. CONCLUSION: These results show that use of unrelated cord blood transplant is an option in patients lacking an HLA identical sibling donor. PMID- 15528144 TI - Graft engineering for allogeneic haploidentical stem cell transplantation. AB - Haploidentical stem cell transplantation has became a clinical reality in the last 10 years as it provides the chance of transplant for about 50% of patients with hematological malignancies who do not have a matched related or unrelated donor. Proper graft preparation for this type of transplant is crucial and this paper analyses our work over the past decade in the search for the optimal graft processing procedure moving from E-rosetting and soybean agglutination, through a combination of negative or positive selection of hematopoietic stem cells to the current method of one-step positive selection. In preparing a graft for haploidentical transplant, three essential requisites must be met. It must contain (1) a megadose (>10 x 10(6) x kg recipient b.w.) of hematopoietic stem cells to overcome the HLA histocompatibility barrier; (2) very few T-lymphocytes (CD3+ cells < 3 x 10(4)/kg recipient b.w.) to prevent severe acute and chronic graft-versus-host disease (GvHD); (3) very few B-lymphocytes to prevent Epstein Barr virus-related lymphoproliferative disorders. With current graft processing technologies based on positive selection of hematopoietic stem cells, these requirements can be met. A 70-80% hematopoietic stem cell recovery ensures the target megadose is achieved in over 70% of cases with a T-cell depletion of more than 4 logs and a B-cell depletion of over 3 logs. Progress in graft processing has ensured primary, sustained engraftment rates of over 90% and has significantly reduced the incidence of severe acute GvHD and EBV-related lymphoproliferative disorders. Modern time-saving automated graft processing devices ensure reproducibility, reliability, and biological safety, which make widespread application of the haploidentical transplant currently feasible. PMID- 15528145 TI - Long-term outcome after haploidentical stem cell transplantation in children. AB - We present an update of our results with transplantation of highly purified stem cells from one to three loci mismatched parental donors. Sixty-three pediatric patients with acute lymphoblastic leukemias (n = 32), acute myeloid, chronic myeloid and myelomonocytic leukemias (n = 13), myelodysplastic syndromes (n = 4), lymphomas (n = 4), and various nonmalignant diseases (n = 10) underwent transplantation. Mobilized peripheral-blood stem cells were selected with either anti-CD34- or anti-CD133-coated microbeads. Patients received a median of 19.5 x 10(6) purified cells and <25,000 CD3+ T lymphocytes per kilogram, with no regular posttransplant pharmacological immunosuppression. Engraftment occurred in 98% of patients (primary sustained engraftment, 83%; engraftment after reconditioning/stem cell boosts, 15%). Moreover, all survivors but one had a stable three-lineage engraftment with a median follow up of 4.1 years (range 0.6 8 years). Primary acute graft-versus-host disease (GvHD) grade II was seen in only 7% of patients. No severe primary acute GvHD grades III-IV occurred. Thirteen percent of the patients developed transient chronic GvHD. Probability of disease-free survival (DFS) at 3 years was 60% for patients with nonmalignant diseases and 48% for patients with acute lymphatic leukemia (ALL)/non-Hodgkin lymphoma (NHL) in complete remission (CR)1-3. None of the ALL/NHL patients with active disease survived. Children with acute and chronic myeloid leukemias had a poorer outcome (3-year DFS = 18%), whereas two of four patients with myelodysplastic syndrome (MDS) are alive. Relapse probability of the whole group was not significantly increased when compared to a historical control group. The incidence of lethal viral infections was 18% between 1995 and 2002 and has since been reduced to 8% by the introduction of new therapeutic strategies. In summary, the use of stem cells from haploidentical parental donors should be strongly considered in all children who need transplantation but lack an identical donor. PMID- 15528147 TI - Immune reconstitution in congenital disorders after HLA-haploidentical hemopoietic stem cell transplantation. PMID- 15528146 TI - WT1-targeted immunotherapy of leukaemia. AB - Since malignant cells are derived from normal cells, many tumour-associated antigens are also expressed in normal tissues. For examples, WT1 is expressed at elevated levels in most leukaemias, but it is also expressed at reduced levels in normal CD34+ haematopoietic stem cells and in progenitor cells of other tissues. Antigen expression in normal tissues is likely to trigger immunological tolerance and thus blunt T cell responses. This could explain the observation that WT1 vaccination in mice frequently fails to stimulate high avidity cytotoxic T cell responses. In order to circumvent tolerance, we have isolated from HLA-A2 negative donors high avidity CTL specific for HLA-A2-presented peptide epitopes of WT1. These allorestricted CTL efficiently kill HLA-A2-positive leukaemia cells but not normal CD34+ haematopoietic stem cells. However, adoptive cellular therapy with allorestricted CTL could only be performed in leukaemia patients rendered tolerant to the infused CTL by prior allogeneic stem cell transplantation. In order to circumvent this limitation, we propose to exploit the TCR of allorestricted CTL as therapeutic tool. TCR gene transfer can be used to take advantage of the specificity of allorestricted CTL and transfer it to patient CTL, while avoiding the transfer of immunogenic alloantigens from the donor CTL to the patient. PMID- 15528148 TI - Hematopoietic stem cell transplantation from alternative sources in adults with high-risk acute leukemia. AB - Since 75% of patients with high-risk acute leukemia do not have a human leukocyte antigen (HLA)-identical sibling, alternative sources for hematopoietic stem cell transplantation (HSCT) are matched unrelated donors (MUD), unrelated umbilical cord blood (UD-UCB) and one HLA haplotype mismatched family members (haploidentical). The chance of finding a suitable donor in the international voluntary donor registries is limited by frequency of the HLA phenotype and the time required to identify the right donor from a potential panel, to establish eligibility and to harvest the cells. In adult MUD recipients, event-free survival ranges up to 50% and refers only to patients who undergo transplant, without taking into account those who do not find a donor. Umbilical cord blood offers the advantages of easy procurement, the absence of risks to donors, the reduced risk of transmitting infections, immediate availability of cryopreserved samples and acceptance of mismatches at two of the six antigens. Although UD-UCB transplantation is a viable option for children, it is seldom considered for adults. The great divergency between body weight and the number of hematopoietic cells in a standard cord blood unit, particularly if associated with a two antigen mismatch, increases the risk of graft failure and delays hematopoietic reconstitution. Work on full-haplotype mismatched transplants has been proceeding for over 20 years. Originally, outcome in leukemia patients was disappointing because of high incidence of severe graft-vs.-host disease in T-replete transplants and high rejection rates in T-cell-depleted transplants. The breakthrough came with the use of a megadose of T-cell-depleted progenitor cells after a high-intensity conditioning regimen. Treating end-stage patients inevitably confounded clinical outcome in the early pilot studies. Today, high risk acute leukemia patients are treated at less advanced stages of disease, receive a reasonably well tolerated conditioning regimen, and benefit from advances in post-transplant immunological reconstitution. All these factors contribute to markedly reduce transplant-related mortality. Overall, event-free survival and transplant-related mortality compare favorably with reports from unrelated matched transplants. T-cell-depleted megadose stem cell transplant from a mismatched family member, who is immediately available, can be offered as a viable option to candidates with high-risk acute leukemias. PMID- 15528149 TI - Generation of transgenic mice expressing human hemoglobin E. AB - Hemoglobin E (HbE, beta26 Glu-->Lys) is the most common abnormal Hb variant in the world, and found in greatest frequency in Southeast (SE) Asia. In the United States, HbE is the third most prevalent variant (after HbS and HbC); and its now increasing frequency is due to immigration from SE Asia. HbE homozygotes present a benign clinical picture, but when HbE is coupled with beta0-thalassemia or HbS, variably severe hemoglobinopathies arise. To date, there are no transgenic animal models of HbE-related diseases. We report here the creation of transgenic mice expressing human HbE as a step toward creating animal models for HbE-related diseases. The betaE mice exhibit red blood cell hypochromia and target cells consistent with those observed in human patients exhibiting HbE trait. Furthermore, the transgenic HbE hemolysates contain increased amounts of Hb oxidation products. PMID- 15528150 TI - The potential of electrospray ionization mass spectrometry for the diagnosis of hemoglobin variants found in newborn screening. AB - Analytical procedures have been developed for the detection and diagnosis of sickle cell disease in newborn babies by analyzing the hemoglobin extracted from dried blood spots on Guthrie cards using electrospray ionization mass spectrometry (ESI-MS). An essential requirement is the ability to reliably differentiate two globin chains whose molecular weights differ by only 1 Da such as adult hemoglobin (Hb A) and Hb C. This has been achieved by improving the accuracy and precision of the molecular weight determination to a fraction of a dalton. We report the potential of mass spectrometry for screening neonates for these debilitating diseases by presenting results from 147 blood spots that had been characterized by phenotypic methods and which include samples from 20 sickle cell disease, 1 beta-thalassemia major, 57 sickle cell trait, and 39 normal babies. In all cases, the mass spectrometric results agreed with the results obtained using conventional analytical practice with high-performance liquid chromatography (HPLC) and isoelectric focusing (IEF). We show that mass spectrometry is a viable technique for the diagnosis of newborns with sickle cell disease or homozygous beta0-thalassemia. PMID- 15528151 TI - Transcriptional potentials of the beta-like globin genes at different developmental stages in transgenic mice and hemoglobin switching. AB - Developmental-stage-specific regulation and physiological levels of expression of the globin genes can be recaptured in transgenic mice carrying a YAC/BAC- or cosmid-based construct. By contrast, proper developmental regulation and high level expression cannot be achieved coordinately in transgenic mice carrying a more manipulated construct, such as a plasmid-based globin gene construct. These differences provide us an opportunity to define the requirements for a developmentally regulated, high-level expression of the globin genes in vivo. To achieve this, as a first step, we studied maximum transcriptional potentials of the beta-globin genes at various stages of development. microLCR-enhanced expression of the epsilon-, gamma-, and beta-globin genes driven by their minimal promoters was estimated and compared with that in betaYAC transgenic mice. Quantitative measurements of steady state mRNA levels of the epsilon-, gamma-, and beta-globin genes showed that the microLCR was able to enhance expression of each beta-like globin gene to levels similar to those in the betaYAC mice. Moreover, transcriptional potentials of each globin gene were unchanged during the entire course of development. These observations indicate that the highest level of expression of the globin genes can be achieved in both embryonic and definitive erythropoiesis regardless of developmental specificity of the genes. This finding implies that transcription suppression is the major mechanism of the developmental specificity of the expression of the beta-like globin genes. PMID- 15528152 TI - Cytochrome P4501A1 polymorphism is associated with susceptibility to acute lymphoblastic leukemia in adult Mexican patients. AB - We studied the role of cytochrome P4501A1 (CYP1A1 Val/Val) genotypes in the etiology of acute lymphoblastic leukemia (ALL) in adult Mexican patients. Distributions of CYP1A1 Val/Val genotypes in peripheral blood DNA samples from 136 healthy controls and 136 adult patients with ALL were evaluated. There was an increased frequency of the CYP1A1 Val/Val genotype among ALL patients, showing a significant association between this genotype and the risk of developing ALL. PMID- 15528153 TI - Report of an International Network of Cancer Treatment and Research workshop on non-Hodgkin's lymphoma in developing countries. AB - The International Network of Cancer Treatment and Research (INCTR) recently organized a workshop on non-Hodgkin lymphomas (NHLs) in selected developing countries with the purpose of examining existing information relating to the pathology and management of these neoplasms, and identifying potential areas for research. This report provides a summary of the information presented and is focused primarily on the pathology of NHLs in children and adults. In most countries, the WHO classification of lymphomas was used and most participating centers included immunohistochemistry using a wide array of lymphoid antibodies as part of routine diagnosis. Some of the series had been reviewed by an external panel of experts. B-cell lymphomas accounted for 82-88% of all NHLs. The proportions of chronic lymphatic leukemia (4-6%), mantle cell lymphoma (MCL, 3 5%), and plasmacytoma (2-4%) were similar in the series presented. However, there was a significant variation in the proportion of follicular lymphoma (FL), which accounted for 15% and 11% in India and Kuwait, but less than 5% in Pakistan and Egypt. All of these frequencies are significantly lower than those reported in Western series. Diffuse large B-cell lymphoma accounted for about 35% of cases in India but for more 50% in other countries, but this difference was not accounted for by an increased incidence in a single lymphoma subtype in India, but rather an apparent paucity of several subtypes (such as mantle cell and marginal zone lymphomas (MZL)) in other series. There were relatively high frequencies of Burkitt lymphoma in Egypt (7%) and precursor T-cell lymphoblastic lymphoma in India (6-7%). Peripheral T-cell lymphomas (PTCLs) (not otherwise specified and angioimmunoblastic subtypes) accounted for 3-5% of NHLs, and extranodal lymphoma of T/NK cell type was rare (<1%). These differences in the relative proportions of NHL subtypes among developing countries and between developing countries and the rest of the world presumably arise from differences in environmental and genetic factors that influence lymphomagenesis and strongly suggest that more research in developing countries would provide valuable insights into the pathogenesis of lymphoid neoplasms. PMID- 15528154 TI - Identification of new mutations of hepcidin and hemojuvelin in patients with HFE C282Y allele. AB - HFE-hemochromatosis is the most common form of hereditary hemochromatosis. The disorder is associated with the homozygous C282Y mutation and has variable phenotype, being modulated by environmental and genetic factors. Candidate modifier genes are hemojuvelin and hepcidin, which are responsible for juvenile hemochromatosis. We used DHPLC to scan mutations in these genes in a cohort of unrelated patients with C282Y mutation. They consisted of 136 C282Y homozygous, 43 heterozygous, and 42 C282Y/H63D compound heterozygous, plus 62 controls subjects. Mutations and polymorphisms were found in 16 patients and 4 controls. Abnormally high indices of iron status were found in subjects C282Y/H63D heterozygous for the N196K hemojuvelin mutation and the -72C > T hepcidin substitution. The already described G71D mutation of hepcidin did not induce evident modification of the C282Y/H63D phenotype. The data show that heterozygous mutations of the hemojuvelin gene contribute like those of hepcidin to the phenotypic heterogeneity of hemochromatosis. However, they are rare and explain only a minor portion of the variable penetrance of the disorder. PMID- 15528155 TI - The mitochondrial nt 16189 polymorphism and hereditary hemochromatosis. AB - It has been claimed that a noncoding mitochondrial polymorphism at nt 16189 is correlated with the penetrance of the homozygous state for the C282Y mutation of the HFE gene. We have genotyped homozygotes for the C282Y mutation and find no relationship between the ferritin levels and the inheritance of the mitochondrial polymorphism. Indeed, the small difference found is in the opposite direction of that reported previously. PMID- 15528156 TI - Anemia and impaired stress-induced erythropoiesis in aceruloplasminemic mice. AB - Ceruloplasmin (Cp) is an abundant, copper-containing plasma protein with an important role in iron homeostasis. Patients with hereditary Cp deficiency have iron deposits in liver and other organs, consistent with impaired iron flux. The mild anemia reported in some patients suggests a possible role for Cp in iron delivery to red cell precursors during erythropoiesis. To investigate this function of Cp, we determined the hematologic parameters in Cp-deficient mice under normal conditions and after erythropoiesis-inducing stress. Cp(-/-) mice have below normal hematocrit, red cell hemoglobin and volume, and serum iron. Red cell number and turnover and reticulocyte counts were identical in Cp(-/-) and Cp(+/+) mice. Thus, Cp(-/-) have mild microcytic, hypochromic anemia consistent with normal red cell formation but defective iron availability. Cp(-/-) and Cp(+/+) mice subjected to phenylhydrazine-induced hemolytic anemia exhibited identical decreases in hematologic parameters, but Cp(-/-) mice showed diminished recovery after removal of the stress. Administration of purified human Cp or iron saturated transferrin to Cp(-/-) mice partially restored hemoglobin formation in reticulocytes. The mild anemia in Cp(-/-) mice and the diminished response to stress may reflect inefficient recycling of iron between the reticuloendothelial and erythropoietic systems. Our findings suggest a role for Cp in erythropoiesis by providing sufficient iron to the erythroid tissue and that the requirement for Cp is raised after erythropoietic stress. PMID- 15528158 TI - Allelic frequency determination of the 24-bp chitotriosidase duplication in the Portuguese population by real-time PCR. AB - Chitotriosidase is a human chitinase produced by macrophages. Its enzymatic activity is markedly elevated in serum of patients suffering from lysosomal storage disorders, as well as other diseases in which macrophages are activated. Therefore, it is a useful tool as a secondary marker in the diagnosis of several disorders including Gaucher disease type 1 and Niemann-Pick disease. The determination of chitotriosidase levels as a diagnosis complement in some lysosomal storage disorders and in enzyme replacement therapy follow-up of Gaucher disease patients is of great importance. However, the fact that a mutation caused by a 24-bp duplication in the CHIT1 gene resulting in deficiency of plasma chitotriosidase activity is very frequent makes the establishment of the frequency of this mutation in different population groups necessary. Furthermore, in order to validate the use of chitotriosidase activity as a marker, it is indispensable to screen individuals for this particular mutation. In this work, we present the results of a study where the allelic frequency of the above mentioned CHIT1 gene mutation was determined in the Portuguese population by real-time PCR. The frequency of carriers encountered in this sample of Portuguese individuals was of 37%. PMID- 15528157 TI - Pharmacology of the human red cell voltage-dependent cation channel. Part II: inactivation and blocking. AB - Pharmacological modulation of the nonselective voltage-dependent cation (NSVDC) channel from human erythrocytes was studied. Using the inorganic cations ruthenium red and La3+, as well as the organic thiol group reagents iodoacetamide (IAA) and N-ethylmaleimide (NEM), it was possible to demonstrate a concentration dependent decrease in the voltage-activated conductance, reflecting an inhibition or inactivation of the channel. Initial voltage activation was achieved by injecting human red cells into sucrose-substituted Ringers with a low chloride concentration, which causes a strongly positive membrane potential to develop, initially determined by the equilibrium potential for Cl- ( approximately +100 mV). Due to the voltage- and time-dependent activation of the cation channel, net effluxes, minimized by addition of a chloride conductance blocker, occurred and Vm gradually decreased and stabilized at a value less positive than E(Cl), reflecting the increased cation conductance, g+, reaching 1.5-2.0 microS/cm2. In the presence of inhibitors of the NSVDC channel, both the membrane potential repolarization and the cation efflux were diminished. PMID- 15528159 TI - Stable long-term gene correction with low-dose radiation conditioning in murine X linked chronic granulomatous disease. AB - We previously demonstrated that low-dose radiation conditioning impairs murine hematopoietic stem cell function, permitting engraftment of syngeneic fresh and transduced marrow cells. In this study, we directly examined the ability of low dose radiation conditioning to permit engraftment of transduced long-term repopulating cells in murine X-linked chronic granulomatous disease (X-CGD), which closely mimics the human disease. X-CGD mice conditioned with 160 cGy were transplanted with 20 x 10(6) MSCV-m91Neo-transduced syngeneic X-CGD marrow cells. The presence of oxidase-positive neutrophils in two independent cohorts of transplanted 160-cGy-conditioned X-CGD recipients was determined by nitroblue tetrazolium testing. Transplanted X-CGD mice (n = 9 total) displayed 1-17% oxidase-positive neutrophils 6-16 months post-transplant. Retroviral marking and NADPH-oxidase-positive neutrophils persisted through serial transplantation, verifying that stem cells were transduced. These results establish that low-dose radiation conditioning results in durable engraftment of low but potentially clinically relevant numbers of functionally reconstituted blood cells in a murine model of X-CGD. PMID- 15528160 TI - Developmental stage-specific expression and tissue distribution of pierisin-1, a guanine-specific ADP-ribosylating toxin, in Pieris rapae. AB - The cabbage butterfly (Pieris rapae) produces pierisin-1, an apoptosis-inducing protein against mammalian cells. In order to clarify the biological role of pierisin-1 in P. rapae, its expression during developmental stages was examined. Low levels of pierisin-1 mRNA and protein were detected in first-instar larvae. During growth until the fifth-instar larval stage, the amounts of the mRNA steadily increased to reach about 50-100 times the initial level. Then it rapidly decreased before pupation. The levels of mRNA in the pupae and the adults were as low as in the first-instar larvae. Levels of pierisin-1 protein also increased around 100 times from the first-instar to the fifth-instar larvae and then gradually decreased by over 90% during the pupal stage. Immunostaining of pierisin-1 demonstrated the protein to be mainly located in fat bodies of fifth instar larvae and early-phase pupae. Although the staining intensity was low, fat bodies of early instars of the larvae and adults were also found to be positive. Moreover, examination of isolated fat body and other tissue samples of the insects were consistent with the above observations. Thus, the results indicate that mRNA of pierisin-1 was highly expressed in late stages of larvae, and that the protein accumulated in fat bodies where it persists during pupation. PMID- 15528162 TI - Effects of antisecretory factor-derived peptides on contractions in guinea pig colon. AB - Two antisecretory factor (AF)-derived peptides have been studied in relation to effects on motility of guinea pig colon. Colon segments were isolated from adult guinea pigs and incubated in Tyrode Ringer. Motility was measured as the force and frequency of contractions upon addition of the derived peptides AF 1 (8 amino acids (aa)) and AF 3 (10 amino acids). At the lowest concentration (5 pM), peptide AF 1 induced a negative effect on the force of contraction in colon segments; an effect that was abolished by the cholinergic agonist carbachol. Peptide AF 3 induced a significant increase in the force of colon contractions at all concentrations (5-180 pM), with carbachol only reducing the effect of peptide AF 3 at a concentration of 15 pM. Both peptides increased contractile frequency, although the overall response was lower for peptide AF 3 than for peptide AF 1. It is concluded that antisecretory factor-derived peptides may play a role in regulating colon motility such that under pathophysiological conditions, they may serve to hasten the evacuation of noxious agents from the large intestine. PMID- 15528161 TI - Dietary calcium deficiency increases Ca2+ uptake and Ca2+ extrusion mechanisms in chick enterocytes. AB - Ca2+ uptake and Ca2+ extrusion mechanisms were studied in enterocytes with different degree of differentiation from chicks adapted to a low Ca2+ diet as compared to animals fed a normal diet. Chicks adapted to a low Ca2+ diet presented hypocalcemia, hypophosphatemia and increased serum 1,25(OH)2D3 and Ca2+ absorption. Low Ca2+ diet increased the alkaline phosphatase (AP) activity, independently of the cellular maturation, but it did not alter gamma-glutamyl transpeptidase activity. Ca2+ uptake, Ca2+-ATPase and Na(+)/Ca2+ exchanger activities and expressions were increased by the mineral-deficient diet either in mature or immature enterocytes. Western blots analysis shows that vitamin D receptor (VDR) expression was much higher in crypt cells than in mature cells. Low Ca2+ diet decreased the number of vitamin D receptor units in both kinds of cells. In conclusion, changes in Ca2+ uptake and Ca2+ extrusion mechanisms in the enterocytes by a low Ca2+ diet appear to be a result of enhanced serum levels of 1,25(OH)2D3, which would promote cellular differentiation producing cells more efficient to express vitamin D dependent genes required for Ca2+ absorption. PMID- 15528163 TI - Novelty stress and reproductive state alters responsiveness to sensory stimuli and 5-HT neuromodulation in crayfish. AB - Sensory stimuli can produce varied responses depending on the physiological state of an animal. Stressors and reproductive stage can result in altered biochemical status that changes the responsiveness of an animal to hormones and neuromodulators, which affects whole animal behavior in relation to sensory stimuli. Crayfish serve as a model for examining the effects of neuromodulators at the neuromuscular junctions (NMJs) and for alterations in stereotypic behaviors for particular stimuli. Thus, we used crayfish to examine the effect of novelty stressors in males and the effect of being gravid in female crayfish to exogenous application of serotonin (5-HT). The responsiveness of neuromuscular junctions to 5-HT revealed that stressed as well as gravid crayfish have a reduced response to 5-HT at NMJs. The stressed crayfish were not fatigued since the basal synaptic responses are large and still showed a pronounced response to 5-HT. Using intact animals to examine a tail flip behavior, we showed that the rate of habituation in tail flipping to a strong repetitive stimulus on the telson is reduced in stressed males. Gravid females show no tail flipping behavior upon telson stimulation. PMID- 15528164 TI - The influence of circadian rhythms on pre- and post-prandial metabolism in the snake Lamprophis fuliginosus. AB - Measuring standard metabolic rate (SMR) and specific dynamic action (SDA) has yielded insight into patterns of energy expenditure in snakes, but less emphasis has been placed on identifying metabolic variation and associated energy cost of circadian rhythms. To estimate SMR, SDA, and identify metabolic variation associated with circadian cycles in nocturnally active African house snakes (Lamprophis fuliginosus), we measured oxygen consumption rates (VO2) at frequent intervals before and during digestion of meals equaling 10%, 20% and 30% of their body mass. Circadian rhythms in metabolism were perceptible in the VO2 data during fasting and after the initial stages of digestion. We estimated SMR of L. fuliginosus (mean mass=16.7+/-0.3 g) to be 0.68+/-0.02 (+/-SEM) mL O2/h at 25 degrees C. Twenty-four hours after eating, VO2 peaked at 3.2-5.3 times SMR. During digestion of meals equaling 10-30% of their body mass, the volume of oxygen consumed ranged from 109 to 119 mL O2 for SMR, whereas extra oxygen consumed for digestion and assimilation ranged from 68 to 256 mL O2 (equivalent to 14.5-17.0% of ingested energy). The oxygen consumed due to the rise in metabolism during the active phase of the daily cycle ranged from 55 to 66 mL O2 during digestion. Peak VO2, digestive scope, and SDA increased with increasing meal size. Comparisons of our estimates to estimates derived from methods used in previous investigations resulted in wide variance of metabolic variables (up to 39%), likely due to the influence of circadian rhythms and activity on the selection of baseline metabolism. We suggest frequent VO2 measurements over multiple days, coupled with mathematical methods that reduce the influence of undesired sources of VO2 variation (e.g., activity, circadian cycles) are needed to reliably assess SMR and SDA in animals exhibiting strong circadian cycles. PMID- 15528165 TI - Sperm storage in males of the snake Crotalus durissus terrificus (Crotalinae: Viperidae) in southeastern Brazil. AB - Seasonal variations in spermatozoa numbers and in sperm motility along the vas deferens in Crotalus durissus terrificus from southeastern Brazil were analyzed. Our data demonstrate storage and motility of the spermatozoa along the vas deferens throughout the year. This is characteristic of a postnuptial reproductive cycle, usually found in snakes living in temperate climates. We describe similarities in reproductive cycle patterns found in the tropical nonhibernator C. durissus terrificus and in hibernator snakes from temperate zones. Our results show that in C. durissus terrificus, a significant difference in spermatozoa counts occurs between winter and summer. Higher numbers of spermatozoa in summer and autumn, due to intense spermiogenesis, coincides with the mating season in autumn. These data indicate that after spermiogenesis in summer, the males combine the peak of sperm storage to the period females are attractive. Mating, however, is not linked to ovulation, and the sperm is stored in the females during winter until fertilization occurs in spring. In the males, after mating, spermatozoon counts low. In spring, they gradually increase, turning again the highest in summer and autumn. During spermiogenesis in the convoluted vas deferens, spermatozoa gain motility, enhancing their performance along their way towards the distal portion. PMID- 15528167 TI - Effects of fasting and exogenous melatonin on annual rhythms in the blue fox (Alopex lagopus). AB - The arctic fox (Alopex lagopus) is a winter-active inhabitant of the high arctic with extreme fluctuations in photoperiod and food availability. The blue fox is a semi-domesticated variant of the wild arctic fox reared for the fur industry. In this study, 48 blue foxes were followed for a year in order to determine the effects of exogenous melatonin and wintertime food deprivation on their reproductive and thyroid axes. Half of the animals were treated with continuous release melatonin capsules in July 2002, and in November-January, the animals were divided into three groups and either fed continuously or fasted for one or two 22-day periods. Food deprivation decreased the plasma triiodothyronine and thyroxine concentrations probably in order to preserve energy due to a decreased metabolic rate. The same was observed in the plasma testosterone levels of the males but not in the plasma estradiol concentrations of the females. Exogenous melatonin advanced the autumn moult and seasonal changes in the voluntary food intake. It also advanced the onset of the testosterone peak in the males. The plasma estradiol levels of the females were unaffected, but the progesterone levels peaked more steeply in the sham-operated females. Melatonin exerted a strong influence not only on the reproductive axis of the males but also on the seasonal food intake. The species seemed quite resistant to periodic involuntary food deprivation. PMID- 15528166 TI - Molecular chaperone function of the Rana catesbeiana small heat shock protein, hsp30. AB - Eukaryotic small heat shock proteins (shps) act as molecular chaperones by binding to denaturing proteins, preventing their heat-induced aggregation and maintaining their solubility until they can be refolded back to their normal state by other chaperones. In this study we report on the functional characterization of a developmentally regulated shsp, hsp30, from the American bullfrog, Rana catesbeiana. An expression vector containing the open reading frame of the hsp30 gene was expressed in Escherichia coli. Purified recombinant hsp30 was recovered as multimeric complexes and was composed of a mixture of alpha-helical and beta-sheet-like structures as determined by circular dichroism analysis. Hsp30 displayed chaperone activity since it inhibited heat-induced aggregation of citrate synthase. Furthermore hsp30 maintained heat-treated luciferase in a folding competent state. For example, heat denatured luciferase when microinjected into Xenopus oocytes did not regain enzyme activity whereas luciferase heat denatured with hsp30 regained 100% enzyme activity. Finally, hsp30 protected the DNA restriction endonuclease, PstI, from heat inactivation. PstI incubated alone at 42 degrees C lost its enzymatic function after 1 h whereas PstI supplemented with hsp30 accurately digested plasmid DNA after 4 h at the elevated temperature. These results clearly indicate a molecular chaperone role for R. catesbeiana hsp30. PMID- 15528168 TI - Effect of dietary vitamin E level on growth, tissue lipid peroxidation, and erythrocyte fragility of transgenic coho salmon, Oncorhynchus kisutch. AB - This study was conducted to investigate the effect of dietary vitamin E concentration on growth performance, iron-catalyzed lipid peroxidation in liver and muscle tissue, and erythrocyte fragility of transgenic growth hormone coho salmon (Oncorhynchus kisutch). Fish were fed one of four isoenergetic and isonitrogenous experimental diets that contained either 11, 29, 50, or 105 IU of vitamin E/kg. Following the 10-week feeding trial, no significant (P>0.05) diet related differences were detected in growth, whole body proximate composition or erythrocyte fragility. The vitamin E contents of liver and muscle, however, were affected by the dietary treatment. Fish fed diets containing > or =50 IU of vitamin E/kg had significantly increased vitamin E concentrations in their tissues. Iron-catalyzed lipid peroxidation of liver and muscle tissue of fish fed elevated dietary vitamin E (> or =50 IU vitamin E/kg diet) was significantly lower (P<0.05) than that noted for fish fed the diet containing no supplemental vitamin E. The results indicated that changes in tissue lipid peroxidation measurements precede clinical signs of sub-optimal vitamin E intake. PMID- 15528169 TI - Cold hardiness abilities vary with the size of the land snail Cornu aspersum. AB - The land snail Cornu aspersum (syn. Helix aspersa) living in Brittany (France) can be considered partially freezing tolerant as it possesses a low ability to supercool and a limited capacity to bear freezing of its body tissues. The absence of a marked cold hardiness strategy permits the emphasis of the role of parameters such as individual size or water mass (W(M)) contained by the organism. Adult snails (shell diameter 30-32 mm) had a supercooling ability, about 1-1.5 degrees C lower than that of immatures (shell diameter 12-20 mm) and survived longer to an exposure to -5 degrees C, with an Lt(50) comprised between 6.0 and 9.8 h against 2.6 to 4.2 h for immature snails. This better ability to bear freezing was explained by the faster dynamic of body ice formation observed in small individuals, which attained ice lethal quantity more rapidly. At the species level, large snails will then tend to be more tolerant to freezing and small ones to be freezing avoidant, a statement also observable at the phylum level. PMID- 15528170 TI - Evidence for placental transfer of lipids during gestation in the viviparous lizard, Pseudemoia entrecasteauxii. AB - During gestation in the viviparous lizard Pseudemoia entrecasteauxii, the fetus obtains nutrients from two sources: uptake of yolk components from the retained egg (lecithotrophy) and transfer of nutrients from the maternal circulation via the placenta (placentotrophy). Although net placentotrophy in this species is indicated by the observation that the neonate contains 1.7 times more dry matter than the egg, the placental transfer of lipid has not been previously demonstrated. Lipid analysis was performed on newly ovulated eggs and on neonates. The weight of total lipid per neonate (8.2+/-0.5 mg) is significantly (P=0.049) greater than that in the egg (6.8+/-0.4 mg), indicating that the placenta must contribute some lipid to the fetus. On the assumption that 50% of the lipid delivered to the fetus from either source is oxidized for energy, it is calculated that the placenta accounts for 58.5% of the fetal lipid requirements, with the remaining 41.5% being derived from the egg. The fatty acid compositions of the triacylglycerol and phospholipid recovered in the neonatal tissue differ substantially from those of the egg. In particular, the proportions of 18:2n-6 and 18:3n-3 are far lower in the neonatal lipids compared with the egg lipids. On the other hand, the proportion of 22:6n-3 in the phospholipid of the neonate is six times higher than in the phospholipid of the egg. The absolute amount (mg) of 22:6n-3 recovered in the total lipid of the neonate is 3.8 times greater than the amount initially present in the egg. By comparison, the amount of total fatty acid in neonatal lipid is 1.2 times greater than the amount in the egg. Thus, there is a preferential use of 22:6n-3 for tissue phospholipid synthesis during development. We conclude that there is net transfer of fatty acids across the placenta to the fetus of P. entrecasteauxii and a high degree of selectivity in the use of the various fatty acids for fetal tissue lipid synthesis. PMID- 15528171 TI - Avenues of extrapulmonary oxygen uptake in western painted turtles (Chrysemys picta belli) at 10 degrees C. AB - The major avenues of extrapulmonary oxygen uptake were determined on submerged western painted turtles (Chrysemys picta bellii) at 10 degrees C by selectively blocking one or more potential pathways for exchange. Previous work indicated that the skin, the cloaca, and the buccopharyngeal cavity can all contribute significantly in various species of turtles. O(2) uptake was calculated from the rate of fall in water P(O(2)) in a closed chamber. Two series of experiments were conducted: in Series 1, each of the potential avenues was mechanically blocked either singly or in combination; in Series 2, active cloacal and buccal pumping were prevented pharmacologically using the paralytic agent rocuronium. In addition in Series 2, N(2)-breathing preceded submergence in some animals and in one set of Series 2 experiments arterial blood was sampled and analyzed for pH, lactate, P(O(2)), and P(CO(2)). Results in both Series 1 and Series 2 revealed that prevention of cloacal and/or buccopharyngeal exchange did not significantly affect total O(2) uptake. Interfering with skin diffusion in Series 1, however, significantly reduced O(2) uptake by 50%. N(2)-breathing prior to submergence in Series 2 did not affect O(2) uptake in paralyzed turtles but significantly increased uptake in unparalyzed turtles without catheters. Blood analysis revealed that all submerged turtles developed lactic acidosis, but the rate of rise in lactate was significantly lower in paralyzed animals. We conclude that passive diffusion through the integument is the principal avenue of aquatic O(2) uptake in this species. PMID- 15528172 TI - Seasonal changes in thermal responses of urban residents to cold exposure. AB - To determine whether urban circumpolar residents show seasonal acclimatisation to cold, thermoregulatory responses and thermal perception during cold exposure were examined in young men during January-March (n=7) and August-September (n=8). Subjects were exposed for 24 h to 22 and to 10 degrees C. Rectal (T(rect)) and skin temperatures were measured throughout the exposure. Oxygen consumption (VO(2)), finger skin blood flow (Q(f)), shivering and cold (CDT) and warm detection thresholds (WDT) were assessed four times during the exposure. Ratings of thermal sensations, comfort and tolerance were recorded using subjective judgement scales at 1-h intervals. During winter, subjects had a significantly higher mean skin temperature at both 22 and 10 degrees C compared with summer. However, skin temperatures decreased more at 10 degrees C in winter and remained higher only in the trunk. Finger skin temperature was higher at 22 degrees C, but lower at 10 degrees C in the winter suggesting an enhanced cold-induced vasoconstriction. Similarly, Q(f) decreased more in winter. The cold detection threshold of the hand was shifted to a lower level in the cold, and more substantially in the winter, which was related to lower skin temperatures in winter. Thermal sensations showed only slight seasonal variation. The observed seasonal differences in thermal responses suggest increased preservation of heat especially in the peripheral areas in winter. Blunted vasomotor and skin temperature responses, which are typical for habituation to cold, were not observed in winter. Instead, the responses in winter resemble aggravated reactions of non-cold acclimatised subjects. PMID- 15528173 TI - Influence of neuropeptide Y (NPY) on food intake and growth of penaeid shrimps Marsupenaeus japonicus and Penaeus semisulcatus (Decapoda: Penaeidae). AB - The effects of neuropeptide Y (NPY) administered intramuscularly or orally on postlarvae (PLs) of two penaeid species were investigated in this study. In experiment 1, food intake (FI) of Marsupenaeus japonicus PLs (0.96 g), injected with NPY at 0.6 microg per g BW, was investigated within 48 h posttreatment. In experiment 2, oral administration of NPY (at doses of 0, 0.125, 0.25, 0.5 microg g(-1) food) on feed intake and growth performance of Penaeus semisulcatus PLs (0.27 g) was examined for 6 weeks. In experiment 1, NPY injection significantly increased average daily FI of M. japonicus PLs within the first 24 h compared to the control (P<0.05), but its stimulatory effect decreased on the second day (P>0.05). The increase in FI was 33% during the first 24 h and 17% during the next 24 h. In experiment 2, significant (P<0.05) differences were found among the groups in terms of weight gain and food utilization (P<0.05). Mean FI significantly increased (as much as 1.3-fold over the control) when NPY was orally administered at doses from 0.125 to 0.5 microg g(-1) feed. There was a positive relationship between FI and final weight (y=-0.972+2.098x, R(2)=0.81) and between FCE and NPY doses in the diets (y=45.37+3.46x, R(2)=0.91). The present findings indicated for the first time that NPY is a potent stimulator of food intake when administered either intramuscularly or orally to penaeid shrimps. PMID- 15528174 TI - Conditioning reduces metabolic rate and time to steady-state in the lizard Naultinus manukanus (Reptilia: Gekkonidae). AB - The rate of oxygen consumption (VO2) is commonly used as a measure of whole organism metabolic rate, but requires the animal to be motionless and at rest. Few studies have measured whether animals that appear motionless are truly at rest, or whether being in a novel environment elevates metabolic rate. We investigated whether conditioning of the gecko Naultinus manukanus to experimental procedures influenced the VO2 and probability of achieving a constant rate of oxygen consumption. Metabolic rate was measured at 24 degrees C in 22 individuals until a steady-state was achieved, or for 80 min if no steady state was reached, once a day on 5 consecutive days (five trials). Geckos in the first trial, when compared with subsequent trials, had a significantly higher mass-adjusted VO2 (0.89+/-0.06 vs. 0.67+/-0.05 ml O2 h(-1), respectively), and time to reach a steady-state VO2 (66+/-8 vs. 47+/-3 min, respectively), as well as a significantly lower probability of reaching a steady-state VO2 (24% vs. 74%, respectively). We conclude that there may be hidden inaccuracies in studies that do not condition animals and that at least one conditioning trial should be used to obtain a metabolic rate at rest for small lizards. PMID- 15528175 TI - Physiological tolerances of the euterrestrial amphipod Arcitalitrus dorrieni (Hunt) as a key to its geographical distribution? A test using mesocosms. AB - An experimental approach was taken to examine the physiological tolerances that may determine the distribution of the euterrestrial amphipod Arcitalitrus dorrieni, a native of Australia which has established itself in the UK over the last 100 years. Two experiments using mesocosms were carried out over consecutive winters to examine the effects of low temperature and leaf litter sodium content on survival. The role of the microhabitat in buffering environmental temperature was also investigated. It was concluded that A. dorrieni may be prevented from colonising areas of Britain by the effects on populations of low winter temperatures and low habitat sodium concentration. There is considerable buffering of environmental temperatures by the leaf litter and soil layers of the microhabitat, and consequently, mortality of A. dorrieni is related to the depth of microhabitat available for it to utilise in avoiding low temperatures. PMID- 15528176 TI - Under "real world" conditions, desflurane increases drug cost without speeding discharge after short ambulatory anesthesia compared to isoflurane. AB - PURPOSE: To compare the measured "real world" perioperative drug cost and recovery associated with desflurane- and isoflurane-based anesthesia in short (less than one hour) ambulatory surgery. METHODS: We conducted a prospective, randomized, blinded trial with patients undergoing arthroscopic meniscectomy under general anesthesia. Following iv induction, patients received either isoflurane (group I; n = 25) or desflurane (group D; n = 20) for maintenance. The primary outcome variable was total perioperative drug cost per patient in Canadian dollars. Secondary outcome variables included volatile agent consumption and cost, adjuvant anesthetic and postanesthesia care unit (PACU) drug cost, readiness for PACU discharge, and incidence of adverse events. RESULTS: Total perioperative drug cost per patient was 14.58 +/- 6.83 Canadian dollars (mean +/- standard deviation) for group I, and 21.47 +/- 5.18 Canadian dollars for group D (P < 0.001). Isoflurane consumption per patient was 6.0 +/- 3.0 mL compared to 18.6 +/- 7.7 mL for desflurane (P < 0.0001); corresponding costs were 0.83 +/- 0.42 Canadian dollars vs 7.61 +/- 3.15 Canadian dollars (P < 0.0001). There were no differences in adjuvant anesthetic or PACU drug cost. All but one patient from each group were deemed ready for PACU discharge at 15 min postoperatively (Aldrete score >or= 9). One patient in group D experienced postoperative nausea. No other adverse events were noted. CONCLUSIONS: Measured total perioperative drug cost for a short ambulatory procedure (less than one hour) under general anesthesia was higher when desflurane rather than isoflurane was used for maintenance, essentially due to volatile agent cost. Desflurane use did not translate into faster PACU discharge under "real world" conditions. PMID- 15528182 TI - Impact of the deltaF508 mutation in first nucleotide-binding domain of human cystic fibrosis transmembrane conductance regulator on domain folding and structure. AB - Cystic fibrosis is caused by defects in the cystic fibrosis transmembrane conductance regulator (CFTR), commonly the deletion of residue Phe-508 (DeltaF508) in the first nucleotide-binding domain (NBD1), which results in a severe reduction in the population of functional channels at the epithelial cell surface. Previous studies employing incomplete NBD1 domains have attributed this to aberrant folding of DeltaF508 NBD1. We report structural and biophysical studies on complete human NBD1 domains, which fail to demonstrate significant changes of in vitro stability or folding kinetics in the presence or absence of the DeltaF508 mutation. Crystal structures show minimal changes in protein conformation but substantial changes in local surface topography at the site of the mutation, which is located in the region of NBD1 believed to interact with the first membrane spanning domain of CFTR. These results raise the possibility that the primary effect of DeltaF508 is a disruption of proper interdomain interactions at this site in CFTR rather than interference with the folding of NBD1. Interestingly, increases in the stability of NBD1 constructs are observed upon introduction of second-site mutations that suppress the trafficking defect caused by the DeltaF508 mutation, suggesting that these suppressors might function indirectly by improving the folding efficiency of NBD1 in the context of the full-length protein. The human NBD1 structures also solidify the understanding of CFTR regulation by showing that its two protein segments that can be phosphorylated both adopt multiple conformations that modulate access to the ATPase active site and functional interdomain interfaces. PMID- 15528183 TI - Production of infectious SIVagm from human cells requires functional inactivation but not viral exclusion of human APOBEC3G. AB - The virus infectivity factor (Vif) is a protein encoded by most primate lentiviruses. Recent evidence suggests that HIV-1 Vif reduces the intracellular levels of the host cytidine deaminase APOBEC3G (Apo3G) and inhibits its packaging into virions. These functions of Vif are thought to be species-specific. Accordingly, HIV-1 Vif can target only human Apo3G (hApo3G), whereas, African green monkey simian immunodeficiency virus (SIVagm) Vif can inhibit African green monkey but not human Apo3G. Consistent with this, we found that SIVagm Vif does not affect the stability of exogenously and endogenously expressed hApo3G and does not prevent packaging of exogenous and endogenous hApo3G into SIVagm virions. Nevertheless, SIVagm Vif supported spreading infection of SIVagm virus in the hApo3G-positive human A3.01 T cell line and rescued infectivity of viruses produced from Apo3G-expressing HeLa cells. Sequence analysis verified that SIVagm Vif inhibited the accumulation of hApo3G-induced mutations, suggesting that SIVagm Vif is indeed active in human cells. Our data suggest that SIVagm Vif can inhibit hApo3G activity without inducing its intracellular degradation or preventing its packaging into virions. PMID- 15528184 TI - Rea1, a dynein-related nuclear AAA-ATPase, is involved in late rRNA processing and nuclear export of 60 S subunits. AB - Rea1, the largest predicted protein in the yeast genome, is a member of the AAA(+) family of ATPases and is associated with pre-60 S ribosomes. Here we report that Rea1 is required for maturation and nuclear export of the pre-60 S subunit. Rea1 exhibits a predominantly nucleoplasmic localization and is present in a late pre-60 S particle together with members of the Rix1 complex. To study the role of Rea1 in ribosome biogenesis, we generated a repressible GAL::REA1 strain and temperature-sensitive rea1 alleles. In vivo depletion of Rea1 results in the significant reduction of mature 60 S subunits concomitant with defects in pre-rRNA processing and late pre-60 S ribosome stability following ITS2 cleavage and prior to the generation of mature 5.8 S rRNA. Strains depleted of the components of the Rix1 complex (Rix1, Ipi1, and Ipi3) showed similar defects. Using an in vivo 60 S subunit export assay, a strong accumulation of the large subunit reporter Rpl25-GFP (green fluorescent protein) in the nucleus and at the nuclear periphery was seen in rea1 mutants at restrictive conditions. PMID- 15528185 TI - Role of serine 10 phosphorylation in p27 stabilization revealed by analysis of p27 knock-in mice harboring a serine 10 mutation. AB - The inhibition of cyclin-dependent kinase activity by p27 contributes to regulation of cell cycle progression. Serine 10 is the major phosphorylation site of p27, and its phosphorylation has been shown to affect the stability and nuclear export of p27 at the G0-G1 transition in transfected cultured cells. To investigate the physiological relevance of p27 phosphorylation on Ser10, we generated p27 "knock-in" mice that harbor an S10A mutation in this protein. Mice homozygous for the mutation (p27(S10A/S10A) mice) were normal in body size, but the abundance of p27 was decreased in many organs, including brain, thymus, spleen, and testis. The stability of p27 in G0 phase was markedly reduced in lymphocytes of p27(S10A/S10A) mice compared with that in wild-type cells, whereas p27 stability in S phase was similar in cells of the two genotypes. The degradation of p27 in cells of the mutant mice at G0 phase was prevented by a proteasome inhibitor. These data indicate that the physiological role of p27 phosphorylation on Ser10 is to stabilize the protein in G0 phase. Unexpectedly, the nuclear export of p27 at the G0-G1 transition occurred normally in p27(S10A/S10A) mouse embryonic fibroblasts, indicating that phosphorylation of Ser10 is dispensable for this process. PMID- 15528186 TI - Expression cloning and demonstration of Enterococcus faecalis lipoamidase (pyruvate dehydrogenase inactivase) as a Ser-Ser-Lys triad amidohydrolase. AB - Enterococcus faecalis lipoamidase was discovered almost 50 years ago (Reed, L. J., Koike, M., Levitch, M. E., and Leach, F. R. (1958) J. Biol. Chem. 232, 143 158) as an enzyme activity that cleaved lipoic acid from small lipoylated molecules and from pyruvate dehydrogenase thereby inactivating the enzyme. Although the partially purified enzyme was a key reagent in proving the crucial role of protein-bound lipoic acid in the reaction mechanism of the 2-oxoacid dehydrogenases, the identity of the lipoamidase protein and the encoding gene remained unknown. We report isolation of the lipoamidase gene by screening an expression library made in an unusual cosmid vector in which the copy number of the vector is readily varied from 1-2 to 40-80 in an appropriate Escherichia coli host. Although designed for manipulation of large genome segments, the vector was also ideally suited to isolation of the gene encoding the extremely toxic lipoamidase. The gene encoding lipoamidase was isolated by screening for expression in E. coli and proved to encode an unexpectedly large protein (80 kDa) that contained the sequence signature of the Ser-Ser-Lys triad amidohydrolase family. The hexa-histidine-tagged protein was expressed in E. coli and purified to near-homogeneity. The purified enzyme was found to cleave both small molecule lipoylated and biotinylated substrates as well as lipoic acid from two 2-oxoacid dehydrogenases and an isolated lipoylated lipoyl domain derived from the pyruvate dehydrogenase E2 subunit. Lipoamidase-mediated inactivation of the 2-oxoacid dehydrogenases was observed both in vivo and in vitro. Mutagenesis studies showed that the residues of the Ser-Ser-Lys triad were required for activity on both small molecule and protein substrates and confirmed that lipoamidase is a member of the Ser-Ser-Lys triad amidohydrolase family. PMID- 15528187 TI - Transcriptional regulation by Lge1p requires a function independent of its role in histone H2B ubiquitination. AB - Saccharomyces cerevisiae cells that have lost their mitochondrial genome (rho(0)) strongly induce transcription of multidrug resistance genes, including the ATP binding cassette transporter gene PDR5. PDR5 induction in rho(0) cells requires the presence of the zinc cluster transcription factor Pdr3p. The PDR3 gene is positively autoregulated in rho(0) cells by virtue of the presence of two binding sites for Pdr3p in its promoter. We identify the novel protein Lge1p as a required participant in the rho(0) activation of PDR3 and PDR5 expression. Lge1p is a nuclear protein that has been found to play a role in ubiquitination of histone H2B at Lys(123). This ubiquitination requires the presence of the ubiquitin-conjugating enzyme Rad6p and the ubiquitin ligase Bre1p. Interestingly, rho(0) strains lacking Lge1p failed to induce PDR3 transcription, but induction was still seen in Deltarad6, Deltabre1, and H2B-K123R mutant strains. Microarray experiments also confirmed that the pattern of gene expression changes seen in cells lacking Lge1p, Bre1p, or Rad6p or containing the H2B-K123R mutant as the only form of H2B share some overlap but are distinct. These findings provide a strong argument that Lge1p has roles in gene regulation independent of its participation in the Rad6p-dependent ubiquitination of H2B. PMID- 15528188 TI - Direct voltage control of signaling via P2Y1 and other Galphaq-coupled receptors. AB - Emerging evidence suggests that Ca2+ release evoked by certain G-protein-coupled receptors can be voltage-dependent; however, the relative contribution of different components of the signaling cascade to this response remains unclear. Using the electrically inexcitable megakaryocyte as a model system, we demonstrate that inositol 1,4,5-trisphosphate-dependent Ca2+ mobilization stimulated by several agonists acting via Galphaq-coupled receptors is potentiated by depolarization and that this effect is most pronounced for ADP. Voltage-dependent Ca2+ release was not induced by direct elevation of inositol 1,4,5-trisphosphate, by agents mimicking diacylglycerol actions, or by activation of phospholipase Cgamma-coupled receptors. The response to voltage did not require voltage-gated Ca2+ channels as it persisted in the presence of nifedipine and was only weakly affected by the holding potential. Strong predepolarizations failed to affect the voltage-dependent Ca2+ increase; thus, an alteration of G protein betagamma subunit binding is also not involved. Megakaryocytes from P2Y1( /-) mice lacked voltage-dependent Ca2+ release during the application of ADP but retained this response after stimulation of other Galphaq-coupled receptors. Although depolarization enhanced Ca2+ mobilization resulting from GTPgammaS dialysis and to a lesser extent during AlF4- or thimerosal, these effects all required the presence of P2Y1 receptors. Taken together, the voltage dependence to Ca2+ release via Galphaq-coupled receptors is not due to control of G-proteins or down-stream signals but, rather, can be explained by a voltage sensitivity at the level of the receptor itself. This effect, which is particularly robust for P2Y1 receptors, has wide-spread implications for cell signaling. PMID- 15528189 TI - Rab13 mediates the continuous endocytic recycling of occludin to the cell surface. AB - During epithelial morphogenesis, adherens junctions (AJs) and tight junctions (TJs) undergo dynamic reorganization, whereas epithelial polarity is transiently lost and reestablished. Although ARF6-mediated endocytic recycling of E-cadherin has been characterized and implicated in the rapid remodeling of AJs, the molecular basis for the dynamic rearrangement of TJs remains elusive. Occludin and claudins are integral membrane proteins comprising TJ strands and are thought to be responsible for establishing and maintaining epithelial polarity. Here we investigated the intracellular transport of occludin and claudins to and from the cell surface. Using cell surface biotinylation and immunofluorescence, we found that a pool of occludin was continuously endocytosed and recycled back to the cell surface in both fibroblastic baby hamster kidney cells and epithelial MTD-1A cells. Biochemical endocytosis and recycling assays revealed that a Rab13 dominant active mutant (Rab13 Q67L) inhibited the postendocytic recycling of occludin, but not that of transferrin receptor and polymeric immunoglobulin receptor in MTD-1A cells. Double immunolabelings showed that a fraction of endocytosed occludin was colocalized with Rab13 in MTD-1A cells. These results suggest that Rab13 specifically mediates the continuous endocytic recycling of occludin to the cell surface in both fibroblastic and epithelial cells. PMID- 15528190 TI - Cyclin-dependent kinase 9 is required for tumor necrosis factor-alpha-stimulated matrix metalloproteinase-9 expression in human lung adenocarcinoma cells. AB - The proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) promotes tumor progression through activation of matrix metalloproteinase (MMP) activity. MMP-9 is a gelatinase secreted by both cancer cells and surrounding stromal cells, and it contributes to TNF-alpha-stimulated tumor invasion and metastasis. Cyclin-dependent kinase 9 (CDK9), the catalytic component of positive transcription elongation factor-b, phosphorylates serine 2 residues in the C terminal domain of RNA polymerase II for productive transcription elongation and is up-regulated upon exposure to various stresses. This study investigated roles of CDK9 in TNF-alpha-stimulated MMP-9 expression in human lung adenocarcinoma cells. CDK9 activity was inhibited using three different strategies, including the CDK9 pharmacological inhibitor 5,6-dichloro-1-beta-D ribofuranosylbenzimidazole (DRB), a dominant-negative CDK9, and a CDK9-specific small interfering RNA. All three approaches reduced TNF-alpha-mediated accumulation of MMP-9 in the conditioned media as demonstrated by gelatin zymography. In contrast, transforming growth factor-beta1-induced accumulation of MMP-2 was unaffected by DRB. Expression of the MMP-9 gene was examined using reverse transcription real time PCR and using a transient transfection assay to evaluate MMP-9 promoter activity. DRB reduced the TNF-alpha-induced increase in MMP-9 mRNA levels but did not effect transforming growth factor-beta1-induced MMP 2 mRNA expression. Consistently DRB and dominant-negative CDK9 completely abrogated TNF-alpha-stimulated human MMP-9 promoter activity. TNF-alpha did not regulate expression or localization of CDK9 or its regulatory partner Cyclin T. However, TNF-alpha stimulated CDK9 binding to Cyclin T and MMP-9 gene occupancy by both CDK9 and the serine 2-phosphorylated form of RNA polymerase II. Our findings indicate that CDK9 mediates TNF-alpha-induced MMP-9 transcription. Disruption of TNF-alpha signaling using CDK9 inhibitors could serve as a potential therapeutic strategy against tumor invasion and metastasis. PMID- 15528191 TI - Interaction between the NH2-terminal domain of eIF4A and the central domain of eIF4G modulates RNA-stimulated ATPase activity. AB - The eukaryotic translation factor 4A (eIF4A) is a member of DEA(D/H)-box RNA helicase family, a diverse group of proteins that couples ATP hydrolysis to RNA binding and duplex separation. eIF4A participates in the initiation of translation by unwinding secondary structure in the 5'-untranslated region of mRNAs and facilitating scanning by the 40 S ribosomal subunit for the initiation codon. eIF4A alone has only weak ATPase and helicase activities, but these are stimulated by eIF4G, eIF4B, and eIF4H. eIF4G has two eIF4A-binding sites, one in the central domain (cp(C3)) and one in the COOH-terminal domain (cp(C2)). In the current work, we demonstrate that these two eIF4G domains have different effects on the RNA-stimulated ATPase activity of eIF4A. cp(C3) stimulates ATP-hydrolytic efficiency by about 40-fold through two mechanisms: lowering K(m)(RNA) by 10-fold and raising k(cat) by 4-fold. cp(C3) also stimulates RNA cross-linking to eIF4A in an ATP-independent manner. Studies with eIF4G and eIF4A variants suggest a model by which cp(C3) alters the conformation of the catalytic site to favor RNA binding. cp(C2) does not stimulate ATPase activity and furthermore increases both K(m)(ATP) (at saturating RNA concentrations) and K(m)(RNA) (at subsaturating ATP concentrations). Both cp(C3) and cp(C2) directly interact with the NH(2)-terminal domain of eIF4A, which possesses conserved ATP- and oligonucleotide-binding motifs, but not with the COOH-terminal domain. PMID- 15528192 TI - Developmental abnormalities of myelin basic protein expression in fyn knock-out brain reveal a role of Fyn in posttranscriptional regulation. AB - Fyn protein-tyrosine kinase (PTK), a member of the Src-PTK family, is essential for myelin development in the central nervous system (CNS). The absence of Fyn activity results in defects in the morphogenesis of oligodendrocyte precursors (OPCs) and CNS hypomyelination. However, molecular mechanisms for Fyn to control CNS myelinogenesis remain elusive. Here we show that Fyn-PTK is significantly up regulated in early OPC differentiation, concentrated in the compact myelin, and declines during myelin development. Despite the high levels of Fyn-PTK expression during early OPC differentiation, Fyn deficiency does not affect the expression of mRNAs that encode myelin structural proteins, including that for the myelin basic protein (MBP), until postnatal day 13 (P13). However, the accumulation rate of MBP mRNA is significantly attenuated during the most active period of myelinogenesis (P13 and P20). Interestingly, the absence of Fyn causes a preferential reduction of the exon-2 containing MBP mRNA isoforms derived from alternative splicing, providing the first evidence that Fyn is required for posttranscriptional regulation of MBP. Consistent with this idea, Fyn phosphorylates the selective RNA-binding protein QKI, which likely modulates the activity of QKI in binding and stabilizing the MBP mRNA. Furthermore, Fyn deficiency exerts an opposing influence on MBP isoform patterning in comparison to that by QKI deficiency. These observations collectively suggest that Fyn plays critical roles in promoting accelerated MBP expression during myelinogenesis in a MBP isoform-preferential manner, and QKI may act in the same pathway downstream of Fyn for MBP mRNA homeostasis. PMID- 15528193 TI - An Inhibitor of the F1 subunit of ATP synthase (IF1) modulates the activity of angiostatin on the endothelial cell surface. AB - Angiostatin binds to endothelial cell (EC) surface F(1)-F(0) ATP synthase, leading to inhibition of EC migration and proliferation during tumor angiogenesis. This has led to a search for angiostatin mimetics specific for this enzyme. A naturally occurring protein that binds to the F1 subunit of ATP synthase and blocks ATP hydrolysis in mitochondria is inhibitor of F1 (IF1). The present study explores the effect of IF1 on cell surface ATP synthase. IF1 protein bound to purified F(1) ATP synthase and inhibited F(1)-dependent ATP hydrolysis consistent with its reported activity in studies of mitochondria. Although exogenous IF1 did not inhibit ATP production on the surface of EC, it did conserve ATP on the cell surface, particularly at low extracellular pH. IF1 inhibited ATP hydrolysis but not ATP synthesis, in contrast to angiostatin, which inhibited both. In cell-based assays used to model angiogenesis in vitro, IF1 did not inhibit EC differentiation to form tubes and only slightly inhibited cell proliferation compared with angiostatin. From these data, we conclude that inhibition of ATP synthesis is necessary for an anti-angiogenic outcome in cell based assays. We propose that IF1 is not an angiostatin mimetic, but it can serve a protective role for EC in the tumor microenvironment. This protection may be overridden in a concentration-dependent manner by angiostatin. In support of this hypothesis, we demonstrate that angiostatin blocks IF1 binding to ATP synthase and abolishes its ability to conserve ATP. These data suggest that there is a relationship between the binding sites of IF1 and angiostatin on ATP synthase and that IF1 could be employed to modulate angiogenesis. PMID- 15528194 TI - Specific structural features of heparan sulfate proteoglycans potentiate neuregulin-1 signaling. AB - Neuregulins are a family of growth and differentiation factors that act through activation of cell-surface erbB receptor tyrosine kinases and have essential functions both during development and on the growth of cancer cells. One alternatively spliced neuregulin-1 form has a distinct heparin-binding immunoglobulin-like domain that enables it to adhere to heparan sulfate proteoglycans at key locations during development and substantially potentiates its activity. We examined the structural specificity needed for neuregulin-1 heparin interactions using a gel mobility shift assay together with an assay that measures the ability of specific oligosaccharides to block erbB receptor phosphorylation in L6 muscle cells. Whereas the N-sulfate group of heparin was most important, the 2-O-sulfate and 6-O-sulfate groups also contributed to neuregulin-1 binding in these two assays. Optimal binding to neuregulin-1 required eight or more heparin disaccharides; however, as few as two disaccharides were still able to bind neuregulin-1 to a lesser extent. The physiological importance of this specificity was shown both by chemical and siRNA treatment of cultured muscle cells. Pretreatment of muscle cells with chlorate that blocks all sulfation or with an siRNA that selectively blocks N-sulfation significantly reduced erbB receptor activation by neuregulin-1 but had no effect on the activity of neuregulin-1 that lacks the heparin-binding domain. These results suggest that the regulation of glycosaminoglycan sulfation is an important biological mechanism that can modulate both the localization and potentiation of neuregulin-1 signaling. PMID- 15528195 TI - A role for rat inositol polyphosphate kinases rIPK2 and rIPK1 in inositol pentakisphosphate and inositol hexakisphosphate production in rat-1 cells. AB - Over 30 inositol polyphosphates are known to exist in mammalian cells; however, the majority of them have uncharacterized functions. In this study we investigated the molecular basis of synthesis of highly phosphorylated inositol polyphosphates (such as inositol tetrakisphosphate, inositol pentakisphosphate (IP5), and inositol hexakisphosphate (IP6)) in rat cells. We report that heterologous expression of rat inositol polyphosphate kinases rIPK2, a dual specificity inositol trisphosphate/inositol tetrakisphosphate kinase, and rIPK1, an IP5 2-kinase, were sufficient to recapitulate IP6 synthesis from inositol 1,4,5-trisphosphate in mutant yeast cells. Overexpression of rIPK2 in Rat-1 cells increased inositol 1,3,4,5,6-pentakisphosphate (I(1,3,4,5,6)P5) levels about 2-3 fold compared with control. Likewise in Rat-1 cells, overexpression of rIPK1 was capable of completely converting I(1,3,4,5,6)P5 to IP6. Simultaneous overexpression of both rIPK2 and rIPK1 in Rat-1 cells increased both IP5 and IP6 levels. To reduce IPK2 activity in Rat-1 cells, we introduced vector-based short interference RNA against rIPK2. Cells harboring the short interference RNA had a 90% reduction of mRNA levels and a 75% decrease of I(1,3,4,5,6)P5. These data confirm the involvement of IPK2 and IPK1 in the conversion of inositol 1,4,5 trisphosphate to IP6 in rat cells. Furthermore these data suggest that rIPK2 and rIPK1 act as key determining steps in production of IP5 and IP6, respectively. The ability to modulate the intracellular inositol polyphosphate levels by altering IPK2 and IPK1 expression in rat cells will provide powerful tools to study the roles of I(1,3,4,5,6)P5 and IP6 in cell signaling. PMID- 15528196 TI - The repressor element silencing transcription factor (REST)-mediated transcriptional repression requires the inhibition of Sp1. AB - The terminal differentiation of neuronal and pancreatic beta-cells requires the specific expression of genes that are targets of an important transcriptional repressor named RE-1 silencing transcription factor (REST). The molecular mechanism by which these REST target genes are expressed only in neuronal and beta-cells and are repressed by REST in other tissues is a central issue in differentiation program of neuronal and beta-cells. Herein, we showed that the transcriptional factor Sp1 was required for expression of most REST target genes both in insulin-secreting cells and neuronal-like cells where REST is absent. Inhibition of REST in a non-beta and a non-neuronal cell model restored the transcriptional activity of Sp1. This activity was also restored by trichostatin A indicating the requirement of histone deacetylases for the REST-mediated silencing of Sp1. Conversely, exogenous introduction of REST blocked Sp1-mediated transcriptional activity. The REST inhibitory effect was mediated through its C terminal repressor domain, which could interact with Sp1. Taken together, these data show that the inhibition of Sp1 by REST is required for the silencing of its target genes expression in non-neuronal and in non-beta-cells. We conclude that the interplay between REST and Sp1 determines the cell-specific expression of REST target genes. PMID- 15528197 TI - Corl1, a novel neuronal lineage-specific transcriptional corepressor for the homeodomain transcription factor Lbx1. AB - During development, neuronal identity is determined by a combination of numerous transcription factors. However, the mechanisms of synergistic action of these factors in transcriptional regulation and subsequent cell fate specification are largely unknown. In this study, we identified a novel gene, Corl1, encoding a nuclear protein with homology to the Ski oncoprotein. Corl1 was highly selectively expressed in the central nervous system (CNS). In the embryonic CNS, Corl1 was expressed in a certain subset of postmitotic neurons generated posterior to the midbrain-hindbrain border. In the developing spinal cord, Corl1 was selectively expressed in the dorsal horn interneurons where a homeodomain transcription factor, Lbx1, is required for proper specification. Corl1 was localized in a nuclear dot-like structure and interacted with general transcriptional corepressors. In addition, Corl1 showed transcriptional repression activity in the GAL4-fusion system, indicating its involvement in the regulation of transcriptional repression. Furthermore, Corl1 interacted with Lbx1 and cooperatively repressed transcription, suggesting that it acts as a transcriptional corepressor for Lbx1 in regulating cell fate determination in the dorsal spinal cord. Corl1 corepressor activity did not depend on Gro/TLE activity, and Gro/TLE also functioned as a corepressor for Lbx1. Thus, Lbx1 can select two independent partners, Corl1 and Gro/TLE, as corepressors. Identification of a novel transcriptional corepressor with neuronal subtype restricted expression might provide insights into the mechanisms of cell fate determination in neurons. PMID- 15528198 TI - Dominant-negative retinoic acid receptors elicit epidermal defects through a non canonical pathway. AB - Previous work has shown that a dominant-negative retinoic acid receptor alpha (dnRARalpha), expressed under the K14 promoter, causes severe epidermal defects. Similar defects are, however, not seen in RARalphagamma double null mutant mice, which lack the entire complement of RARs expressed in the epidermis. To investigate the mechanism of action of these dominant-negative receptors, dnRARalpha or a DNA binding-deficient variant, dnRARalpha(DBD), were targeted to the basal epidermis. Expression of either receptor type led to similar epidermal phenotypes suggesting that both RAR mutants acted through a common mechanism. The epidermal phenotype was reminiscent of defects seen in p63(-/-) mice. Consistent with this, reduced p63 expression was observed in transgenic offspring expressing either RAR mutant, suggesting that down-regulation of p63 might underlie the effects of these receptors on epidermal development. By contrast, expression of p63 in the epidermis of RARalphagamma(-/-) offspring was unaffected, indicating that RARs were not essential for p63 expression. These findings suggest that dnRARs may impact on epidermal development through one or more non-canonical pathways, which are independent of receptor-DNA interaction. PMID- 15528199 TI - Altered interaction and expression of proteins involved in neurosecretion in scrapie-infected GT1-1 cells. AB - Prions cause transmissible and fatal diseases that are associated with spongiform degeneration, astrogliosis, and loss of axon terminals in the brains. To determine the expression of proteins involved in neurosecretion and synaptic functions after prion infection, gonadotropin-releasing hormone neuronal cell line subclone (GT1-1) was infected with the RML scrapie strain and analyzed by Western blotting, real time PCR, and immunohistochemistry. As revealed by Western blotting of lysates exposed to different temperatures, the levels of complexed SNAP-25, syntaxin 1A, and synaptophysin were decreased in scrapie-infected GT1-1 cells (ScGT1-1), whereas the level of monomeric forms of these proteins was increased and correlated to the level of scrapie prion protein (PrPSc). However, when complex formation was prevented by prolonged heating of samples in SDS, the levels of monomeric SNAP-25, syntaxin 1A and synaptophysin in ScGT1-1 cells were decreased in comparison to GT1-1 cells. The reduced level of SNAP-25 was observed as early as 32 days postinfection. Increased mRNA levels of both splice variants SNAP-25a and -b in ScGT1-1 cells were seen. No difference in the morphology, neuritic outgrowth or distribution of SNAP-25, syntaxin 1A, or synaptophysin could be observed in ScGT1-1 cells. Treatment with quinacrine or pentosan polysulfate cleared the PrPSc from the ScGT1-1 cell cultures, and the increase in levels of monomeric SNAP-25 and synaptophysin was reversible. These results indicate that a scrapie infection can cause changes in the expression of proteins involved in neuronal secretion, which may be of pathogenetic relevance for the axon terminal changes seen in prion-infected brains. PMID- 15528200 TI - Characterization of a second Arabidopsis thaliana prolyl 4-hydroxylase with distinct substrate specificity. AB - 4-Hydroxyproline is found in collagens, collagen-like proteins, elastin, and the hypoxia-inducible transcription factor in animals and in many hydroxyproline-rich glycoproteins in plants. We report here on the cloning and characterization of a second plant P4H (prolyl 4-hydroxylase), At-P4H-2, from Arabidopsis thaliana. It consists of 299 amino acids and shows 33% sequence identity to the first characterized isoenzyme, At-P4H-1. A characteristic feature of the At-P4H-2 polypeptide is a 49-amino-acid C-terminal toxin homology domain with 6 cysteines that is not found in At-P4H-1 but is present in a putative rice P4H homologue. At P4H-2 differed distinctly from At-P4H-1 in its substrate specificity. Recombinant At-P4H-2 hydroxylated poly(L-proline) and extensin and arabinogalactan-like peptides effectively but with much higher Km values than At-P4H-1, suggesting different roles for the two At-P4Hs in the plant cell. Unlike At-P4H-1, At-P4H-2 hydroxylated collagen-like peptides only very inefficiently and did not hydroxylate hypoxia-inducible transcription factor alpha-like peptides at all. All the peptides efficiently hydroxylated by At-P4H-2 had at least 3 consecutive prolines, suggesting that these may represent a minimum requirement for efficient hydroxylation by this isoenzyme. N-terminal sequencing of an extensin-like peptide SPPPVYKSPPPPVKHYSPPPV indicated that At-P4H-2 preferentially hydroxylated the 3rd proline in the C-terminal PPP triplet. The Km values of At-P4H-2 for the reaction cosubstrates Fe2+, 2-oxoglutarate, and ascorbate were similar to those of At-P4H-1 with the exception that the Km for iron was about 3-fold lower. Pyridine-2,4-dicarboxylate and pyridine-2,5-dicarboxylate, well known competitive inhibitors of the vertebrate P4Hs with respect to 2-oxoglutarate, were also competitive inhibitors of At-P4H-2 but with Ki values 5-100-fold higher than those of human type I collagen P4H. It thus seems that there are some distinct differences in the structure of the 2-oxoglutarate-binding site between At-P4H-2 and the animal collagen P4Hs. PMID- 15528201 TI - Regional specificity of human ether-a'-go-go-related gene channel activation and inactivation gating. AB - Slow activation and rapid C-type inactivation produce inward rectification of the current-voltage relationship for human ether-a'-go-go-related gene (hERG) channels. To characterize the voltage sensor movement associated with hERG activation and inactivation, we performed an Ala scan of the 32 amino acids (Gly(514)-Tyr(545)) that comprise the S4 domain and the flanking S3-S4 and S4-S5 linkers. Gating and ionic currents of wild-type and mutant channels were measured using cut-open oocyte Vaseline gap and two microelectrode voltage clamp techniques to determine the voltage dependence of charge movement, activation, and inactivation. Mapping the position of the charge-perturbing mutations (defined as |DeltaDeltaG| > 1.0 kcal/mol) on a three-dimensional S4 homology model revealed a spiral pattern. As expected, mutation of these residues also altered activation. However, mutation of residues in the S3-S4 and S4-S5 linkers and the C-terminal end of S4 perturbed activation (|DeltaDeltaG| > 1.0 kcal/mol) without altering charge movement, suggesting that the native residues in these regions couple S4 movement to the opening of the activation gate or stabilize the open or closed state of the channel. Finally, mutation of a distinct set of residues impacted inactivation and mapped to a single face of the S4 helix that was devoid of activation-perturbing residues. These results define regions on the S4 voltage sensor that contribute differentially to hERG activation and inactivation gating. PMID- 15528202 TI - Pathological prion protein exposure switches on neuronal mitogen-activated protein kinase pathway resulting in microglia recruitment. AB - Transmissible spongiform encephalopathies are accompanied by the recruitment of microglial cells in the vicinity of amyloid aggregates of the pathological prion protein (PrPres). We previously showed that PrPres itself triggered the recruitment of microglia by interacting with neurons leading to the up-regulation of the expression level of chemokines, mainly RANTES (regulated on activation normal T cell expressed and secreted). The intracellular mechanisms underlying the PrPres-inducible expression of chemokines in this setting are not clear. Here we demonstrate that the mitogen-activated protein kinase pathway is switched on shortly after PrPres exposure to neurons leading to the expression of early growth response factor-1 (Egr-1), a transcription factor initially linked to differentiation and growth and to up-regulation of RANTES mRNA expression. PD98059, a selective inhibitor of extracellular signal-regulated kinase1/2 activation, resulted in a decrease of RANTES mRNA expression and as a consequence to the lowering of microglial cell migration. Neuronal overexpression of Nab2, a corepressor of Egr-1, produced similar effects. PrPres-induced chemoattraction is independent of the presence of PrPc and the laminin receptor on the neuronal cell surface. Our report is the first demonstration that PrPres exposure on neurons results in the activation of the MAP kinase signaling pathway that acts as a master switch to trigger neuronal expression of regulators of chemoattraction. PMID- 15528203 TI - Regulation of ATP-sensitive potassium channel subunit Kir6.2 expression in rat intestinal insulin-producing progenitor cells. AB - We have reported that the combined expression of Pdx-1 (pancreatic duodenal homeobox 1) and Isl-1 (islet 1) enables immature rat enterocytes (IEC-6) to produce and release insulin. A key component regulating the release of insulin is the ATP-sensitive potassium channel subunit Kir6.2. To investigate the regulation of Kir6.2 gene expression, we assessed Kir6.2 expression in IEC-6 cells expressing Pdx-1 and/or Isl-1. We observed that Kir6.2 protein was expressed de novo in IEC-6 cells expressing both Pdx-1 and Isl-1 but not in cells expressing Pdx-1 alone. Next, we analyzed the regions of the Kir6.2 promoter (-1677/-45) by performing a luciferase assay and electrophoretic mobility shift assay. The results have demonstrated that Kir6.2 promoter possesses two regions regulating the promoter activity: a Foxa2-binding site (-1364 to -1210) and an Sp1/Sp3 binding site (-1035 to -939). The additional expression of Isl-1 in IEC-6 cells expressing Pdx-1 attenuated overexpression of Foxa2 protein and enhanced Kir6.2 expression. Finally, knockdown of Isl-1 using the iRNA technique resulted in decreased expression of Kir6.2 protein in a rat pancreatic beta-cell line (RIN-5F cells). These results indicate that expression of Kir6.2 in the rat intestine is moderated by Isl-1. PMID- 15528204 TI - Amyloid-beta protofibrils differ from amyloid-beta aggregates induced in dilute hexafluoroisopropanol in stability and morphology. AB - The brains of Alzheimer's disease (AD) patients contain large numbers of amyloid plaques that are rich in fibrils composed of 40- and 42-residue amyloid-beta (Abeta) peptides. Several lines of evidence indicate that fibrillar Abeta and especially soluble Abeta aggregates are important in the etiology of AD. Recent reports also stress that amyloid aggregates are polymorphic and that a single polypeptide can fold into multiple amyloid conformations. Here we demonstrate that Abeta-(1-40) can form soluble aggregates with predominant beta-structures that differ in stability and morphology. One class of aggregates involved soluble Abeta protofibrils, prepared by vigorous overnight agitation of monomeric Abeta (1-40) at low ionic strength. Dilution of these aggregation reactions induced disaggregation to monomers as measured by size exclusion chromatography. Protofibril concentrations monitored by thioflavin T fluorescence decreased in at least two kinetic phases, with initial disaggregation (rate constant approximately 1 h(-1)) followed by a much slower secondary phase. Incubation of the reactions without agitation resulted in less disaggregation at slower rates, indicating that the protofibrils became progressively more stable over time. In fact, protofibrils isolated by size exclusion chromatography were completely stable and gave no disaggregation. A second class of soluble Abeta aggregates was generated rapidly (<10 min) in buffered 2% hexafluoroisopropanol (HFIP). These aggregates showed increased thioflavin T fluorescence and were rich in beta structure by circular dichroism. Electron microscopy and atomic force microscopy revealed initial globular clusters that progressed over several days to soluble fibrous aggregates. When diluted out of HFIP, these aggregates initially were very unstable and disaggregated completely within 2 min. However, their stability increased as they progressed to fibers. Relative to Abeta protofibrils, the HFIP induced aggregates seeded elongation by Abeta monomer deposition very poorly. The techniques used to distinguish these two classes of soluble Abeta aggregates may be useful in characterizing Abeta aggregates formed in vivo. PMID- 15528205 TI - Roles of distal Asp in heme oxygenase from Corynebacterium diphtheriae, HmuO: A water-driven oxygen activation mechanism. AB - Heme oxygenases found in mammals, plants, and bacteria catalyze degradation of heme using the same mechanism. Roles of distal Asp (Asp-136) residue in HmuO, a heme oxygenase of Corynebacterium diphtheriae, have been investigated by site directed mutagenesis, enzyme kinetics, resonance Raman spectroscopy, and x-ray crystallography. Replacements of the Asp-136 by Ala and Phe resulted in reduced heme degradation activity due to the formation of ferryl heme, showing that the distal Asp is critical in HmuO heme oxygenase activity. D136N HmuO catalyzed heme degradation at a similar efficiency to wild type and D136E HmuO, implying that the carboxylate moiety is not required for the heme catabolism by HmuO. Resonance Raman results suggest that the inactive ferryl heme formation in the HmuO mutants is induced by disruption of the interaction between a reactive Fe-OOH species and an adjacent distal pocket water molecule. Crystal structural analysis of the HmuO mutants confirms partial disappearance of this nearby water in D136A HmuO. Our results provide the first experimental evidence for the catalytic importance of the nearby water molecule that can be universally critical in heme oxygenase catalysis and propose that the distal Asp helps in positioning the key water molecule at a position suitable for efficient activation of the Fe-OOH species. PMID- 15528206 TI - c-Jun N-terminal kinase 3 deficiency protects neurons from axotomy-induced death in vivo through mechanisms independent of c-Jun phosphorylation. AB - Both the transcription factor c-Jun and the c-Jun N-terminal kinases (JNKs) have been associated with neuronal loss in several death paradigms. JNK are key regulators of c-Jun and a common accepted model has been that JNKs mediate neuronal death through modulation of c-Jun activation. In the present study, we examined whether JNK2 and -3 (JNK members most associated with neuronal loss) deficiency can rescue neuronal loss caused by facial and sciatic nerve axotomy in the neonate in vivo. JNK2, JNK3, and JNK2/3 double-deficient neurons displayed significantly less death in the facial nerves of the CNS when compared with controls. JNK2 and JNK2/3 double-deficient animals also showed reduced c-Jun phosphorylation and induction following axotomy, consistent with the model that JNK acts to regulate death by activating c-Jun. Of significance, however, protection of facial nerves in JNK3-deficient animals was not accompanied by reduction in c-Jun activation. These results suggest that JNKs can mediate death independently of c-Jun. Importantly, the lack of correlation between JNK3 deficiency and c-Jun induction was not universal. In a sciatic axotomy model of neuronal injury in the neonate, death of DRG neurons was also reduced by JNK3 deficiency. However, in this case, c-Jun activation was also eliminated. PMID- 15528207 TI - STAT5B-mediated growth hormone signaling is organized by highly dynamic microtubules in hepatic cells. AB - In the last decade, the notion that microtubules are critical to the spatial organization of signal transduction and contribute to the transmission of signals to downstream targets has been proposed. Because the STAT5B transduction and transcription factor is the major STAT protein activated by growth hormone stimulation in hepatocytes and is a crossroads between many signaling pathways, we studied the involvement of microtubules in STAT5B-mediated growth hormone signaling pathway in the highly differentiated and polarized WIF-B hepatic cell line. We showed that depolymerization of the microtubule network impaired STAT5B translocation to the nucleus upon growth hormone treatment. A significant amount of STAT5B binds to microtubules, while STAT5A and STAT3 are exclusively compartmentalized in the cytosol. Moreover, taxol-induced stabilization of microtubules released STAT5B from its binding, and we show that STAT5B binds specifically to the highly dynamic microtubules and is absent of the stable microtubule subpopulation. The specific involvement of dynamic microtubule subpopulation in growth hormone signaling pathway was confirmed by the inhibition of growth hormone-induced STAT5B nuclear translocation after stabilization of microtubules or specific disruption of highly dynamic microtubules. Upon growth hormone treatment, MT-bound STAT5B was rapidly released from microtubules by a dynein-dependent transport to the nucleus. Altogether, our findings indicate that the labile microtubule subpopulation specifically and dynamically organizes STAT5B-mediated growth hormone signaling in hepatic cells. PMID- 15528208 TI - Characterization of the interaction between retinoic acid receptor/retinoid X receptor (RAR/RXR) heterodimers and transcriptional coactivators through structural and fluorescence anisotropy studies. AB - Retinoid receptors (RARs and RXRs) are ligand-activated transcription factors that regulate the transcription of target genes by recruiting coregulator complexes at cognate promoters. To understand the effects of heterodimerization and ligand binding on coactivator recruitment, we solved the crystal structure of the complex between the RARbeta/RXRalpha ligand-binding domain heterodimer, its 9 cis retinoic acid ligand, and an LXXLL-containing peptide (termed NR box 2) derived from the nuclear receptor interaction domain (NID) of the TRAP220 coactivator. In parallel, we measured the binding affinities of the isolated NR box 2 peptide or the full-length NID of the coactivator SRC-1 for retinoid receptors in the presence of various types of ligands. Our correlative analysis of three-dimensional structures and fluorescence data reveals that heterodimerization does not significantly alter the structure of individual subunits or their intrinsic capacity to interact with NR box 2. Similarly, we show that the ability of a protomer to recruit NR box 2 does not vary as a function of the ligand binding status of the partner receptor. In contrast, the strength of the overall association between the heterodimer and the full-length SRC-1 NID is dictated by the combinatorial action of RAR and RXR ligands, the simultaneous presence of the two receptor agonists being required for highest binding affinity. We identified an LXXLL peptide-driven mechanism by which the concerted reorientation of three phenylalanine side chains generates an "aromatic clamp" that locks the RXR activation helix H12 in the transcriptionally active conformation. Finally, we show how variations of helix H11-ligand interactions can alter the communication pathway linking helices H11, H12, and the connecting loop L11-12 to the coactivator-binding site. Together, our results reveal molecular and structural features that impact on the ligand-dependent interaction of the RAR/RXR heterodimer with nuclear receptor coactivators. PMID- 15528209 TI - Microtubule-associated protein 1S, a short and ubiquitously expressed member of the microtubule-associated protein 1 family. AB - The related high molecular mass microtubule-associated proteins (MAPs) MAP1A and MAP1B are predominantly expressed in the nervous system and are involved in axon guidance and synaptic function. MAP1B is implicated in fragile X mental retardation, giant axonal neuropathy, and ataxia type 1. We report the functional characterization of a novel member of the microtubule-associated protein 1 family, which we termed MAP1S (corresponding to sequence data bank entries for VCY2IP1 and C19ORF5). MAP1S contains the three hallmark domains of the microtubule-associated protein 1 family but hardly any additional sequences. It decorates neuronal microtubules and copurifies with tubulin from brain. MAP1S is synthesized as a precursor protein that is partially cleaved into heavy and light chains in a tissue-specific manner. Heavy and light chains interact to form the MAP1S complex. The light chain binds, bundles, and stabilizes microtubules and binds to actin. The heavy chain appears to regulate light chain activity. In contrast to MAP1A and MAP1B, MAP1S is expressed in a wide range of tissues in addition to neurons and represents the non-neuronal counterpart of this cytolinker family. PMID- 15528210 TI - Nicotinic acid adenine dinucleotide phosphate potentiates neurite outgrowth. AB - Ca(2+) regulates a spectrum of cellular processes including many aspects of neuronal function. Ca(2+)-sensitive events such as neurite extension and axonal guidance are driven by Ca(2+) signals that are precisely organized in both time and space. These complex cues result from both Ca(2+) influx across the plasma membrane and the mobilization of intracellular Ca(2+) stores. In the present study, using rat cortical neurons, we have examined the effects of the novel intracellular Ca(2+)-mobilizing messenger nicotinic acid adenine dinucleotide phosphate (NAADP) on neurite length and cytosolic Ca(2+) levels. We show that NAADP potentiates neurite extension in response to serum and nerve growth factor and stimulates increases in cytosolic Ca(2+) from bafilomycin-sensitive Ca(2+) stores. Simultaneous blockade of inositol trisphosphate and ryanodine receptors abolished the effects of NAADP on neurite length and reduced the magnitude of NAADP-mediated Ca(2+) signals. This is the first report demonstrating functional NAADP receptors in a mammalian neuron. Interplay between NAADP receptors and more established intracellular Ca(2+) channels may therefore play important signaling roles in the nervous system. PMID- 15528211 TI - A novel C-terminal motif is necessary for the export of the vasopressin V1b/V3 receptor to the plasma membrane. AB - Little is known about endoplasmic reticulum (ER) export signals, particularly those of members of the G-protein-coupled receptor family. We investigated the structural motifs involved in membrane export of the human pituitary vasopressin V1b/V3 receptor. A series of V3 receptors carrying deletions and point mutations were expressed in AtT20 corticotroph cells. We analyzed the export of these receptors by monitoring radioligand binding and by analysis of a V3 receptor tagged with both green fluorescent protein and Myc epitopes by a novel flow cytometry-based method. This novel method allowed us to quantify total and membrane-bound receptor expression. Receptors lacking the C terminus were not expressed at the cell surface, suggesting the presence of an export motif in this domain. The distal C terminus contains two di-acidic (DXE) ER export motifs; however, mutating both these motifs had no effect on the V3 receptor export. The proximal C terminus contains a di-leucine (345)LL(346) motif surrounded by the hydrophobic residues Phe(341), Asn(342), and Leu(350). The mutation of one or more of these five residues abolished up to 100% of the receptor export. In addition, these mutants colocalized with calnexin, demonstrating that they were retained in the ER. Finally, this motif was sufficient to confer export properties on a CD8alpha glycoprotein-V3 receptor chimera. In conclusion, we have identified a novel export motif, FN(X)(2)LL(X)(3)L, in the C terminus of the V3 receptor. PMID- 15528212 TI - Myc antagonizes Ras-mediated growth arrest in leukemia cells through the inhibition of the Ras-ERK-p21Cip1 pathway. AB - Even though RAS usually acts as a dominant transforming oncogene, in primary fibroblasts and some established cell lines Ras inhibits proliferation. This can explain the virtual absence of RAS mutations in some types of tumors, such as chronic myeloid leukemia (CML). We report that in the CML cell line K562 Ras induces p21Cip1 expression through the Raf-MEK-ERK pathway. Because K562 cells are deficient for p15INK4b, p16INK4a, p14ARF, and p53, this would be the main mechanism whereby Ras up-regulates p21 expression in these cells. Accordingly, we also found that Ras suppresses K562 growth by signaling through the Raf-ERK pathway. Because c-Myc and Ras cooperate in cell transformation and c-Myc is up regulated in CML, we investigated the effect of c-Myc on Ras activity in K562 cells. c-Myc antagonized the induction of p21Cip1 mediated by oncogenic H-, K-, and N-Ras and by constitutively activated Raf and ERK2. Activation of the p21Cip1 promoter by Ras was dependent on Sp1/3 binding sites in K562. However, mutational analysis of the p21 promoter and the use of a Gal4-Sp1 chimeric protein strongly suggest that c-Myc affects Sp1 transcriptional activity but not the binding of Sp1 to the p21 promoter. c-Myc-mediated impairment of Ras activity on p21 expression required a transactivation domain, a DNA binding region, and a Max binding region. Moreover, the effect was independent of Miz1 binding to c-Myc. Consistent with its effect on p21Cip1 expression, c-Myc rescued cell growth inhibition induced by Ras. The data suggest that in particular tumor types, such as those associated with CML, c-Myc contributes to tumorigenesis by inhibiting Ras antiproliferative activity. PMID- 15528213 TI - Evolutionarily conserved multiple C2 domain proteins with two transmembrane regions (MCTPs) and unusual Ca2+ binding properties. AB - C2 domains are primarily found in signal transduction proteins such as protein kinase C, which generally contain a single C2 domain, and in membrane trafficking proteins such as synaptotagmins, which generally contain multiple C2 domains. In both classes of proteins, C2 domains usually regulate the respective protein's function by forming Ca(2+)-dependent or Ca(2+)-independent phospholipid complexes. We now describe MCTPs (multiple C2 domain and transmembrane region proteins), a novel family of evolutionarily conserved C2 domain proteins with unusual Ca(2+)-dependent properties. MCTPs are composed of a variable N-terminal sequence, three C2 domains, two transmembrane regions, and a short C-terminal sequence. The invertebrate organisms Caenorhabditis elegans and Drosophila melanogaster express a single MCTP gene, whereas vertebrates express two MCTP genes (MCTP1 and MCTP2) whose primary transcripts are extensively alternatively spliced. Most of the MCTP sequences, in particular the C2 domains, are highly conserved. All MCTP C2 domains except for the second C2 domain of MCTP2 include a perfect Ca2+/phospholipid-binding consensus sequence. To determine whether the C2 domains of MCTPs actually function as Ca2+/phospholipid-binding modules, we analyzed their Ca2+ and phospholipid binding properties. Surprisingly, we found that none of the three MCTP1 C2 domains interacted with negatively charged or neutral phospholipids in the presence or absence of Ca2+. However, Ca2+ titrations monitored via intrinsic tryptophan fluorescence revealed that all three C2 domains bound Ca2+ in the absence of phospholipids with a high apparent affinity (EC50 of approximately 1.3-2.3 microM). Our data thus reveal that MCTPs are evolutionarily conserved C2 domain proteins that are unusual in that the C2 domains are anchored in the membrane by two closely spaced transmembrane regions and represent Ca(2+)-binding but not phospholipid-binding modules. PMID- 15528214 TI - Multiple gestation and infertility treatment: registration, reflection and reaction--the Belgian project. AB - Multiple pregnancies associated with infertility treatment are recognized as an adverse outcome and are responsible for morbidity and mortality related to prematurity and very low birthweight population. Due to the epidemic of iatrogenic multiple births, the incidence of maternal, perinatal and childhood morbidity and mortality has increased. This results in a hidden healthcare cost of infertility therapy and this may lead to social and political concern. Reducing the number of embryos transferred and the use of natural cycle IVF will surely decrease the number of multiple gestations. Consequently, optimized cryopreservation programmes will be essential. For non-IVF hormonal stimulation, responsible for more than one-third of all multiple pregnancies after infertility treatment, a strict ovarian stimulation protocol aiming at mono-ovulation is crucial. Multifetal pregnancy reduction is an effective method to reduce high order multiplets but carries its own risk of medical and emotional complications. Excellent data collection of all infertility treatments is needed in our discussion with policy makers. The Belgian project, in which reimbursement of assisted reproduction technology-related laboratory activities is linked to a transfer policy aiming at substantial multiple pregnancy reduction, is a good example of cost-efficient health care through responsible, well considered clinical practice. PMID- 15528215 TI - Gene expression profiling of chemically induced rat mammary gland cancer. AB - Exposure to carcinogens through diet, the atmosphere and other means is generally regarded as influencing human cancer risk, but the impact of specific environmental carcinogens on human breast cancer incidence is still unknown. We examined whether distinct chemical carcinogens induce a unique transcriptional profile in mammary gland cancer that is characteristic of the etiologic agent. Rat mammary gland cancers (n = 34) were generated by various carcinogens, including the food-derived heterocyclic amines 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 7,12-dimethylbenz[a]anthracene, N-nitrosomethylurea and 4-aminobiphenyl. The histopathology of the carcinomas was graded using a modified Scarff-Bloom Richardson scheme and the gene expression profiles in the carcinomas were evaluated on a 10K cDNA microarray. Unsupervised hierarchical clustering analysis revealed two major clusters of carcinomas irrespective of the carcinogenic agent that distinguished two groups with different histopathological parameters (degree of differentiation, nuclear grade, mitotic activity, epithelial cell growth pattern and necrosis). Using class comparison analysis and hierarchical clustering of all carcinomas irrespective of histopathology, gene expression profiles were further shown to be statistically differentially expressed according to the carcinogenic agent. These findings indicate that the transcriptional program in carcinomas is unique to the etiologic agent and can be observed among a diverse set of carcinogens despite variations in carcinoma histopathology. The ability to use microarray analysis to discern an etiology specific profile among a pathologically heterogeneous group of breast carcinomas may ultimately be valuable in determining the role of environmental chemical carcinogens in human breast cancer risk. PMID- 15528216 TI - Peptides specific to the galectin-3 carbohydrate recognition domain inhibit metastasis-associated cancer cell adhesion. AB - Intravascular cancer cell adhesion plays a significant role in the metastatic process. Studies indicate that galectin-3, a member of the galectin family of soluble animal lectins, is involved in carbohydrate-mediated metastatic cell heterotypic (between carcinoma cells and endothelium) and homotypic (between carcinoma cells) adhesion via interactions with the tumor-specific Thomsen Friedenreich glycoantigen (TFAg). We hypothesized that blocking the galectin-3 carbohydrate recognition domain with synthetic peptides would significantly reduce metastasis-associated carcinoma cell adhesion. To test this hypothesis, we identified peptide antagonists of the galectin-3 carbohydrate recognition domain using combinatorial bacteriophage display technology. The peptides bound with high affinity to purified recombinant galectin-3 protein (K(d) approximately 17 80 nM) and to cell surface galectin-3. Experiments with a series of recombinant serially truncated galectin-3 mutants indicated that the peptides bound the carbohydrate recognition domain of galectin-3. Furthermore, the peptides did not bind the carbohydrate recognition domain of other galectins and plant lectins. Synthetic galectin-3 carbohydrate recognition domain-specific peptides blocked the interaction between galectin-3 and TFAg and significantly inhibited rolling and stable heterotypic adhesion of human MDA-MB-435 breast carcinoma cells to endothelial cells under flow conditions, as well as homotypic tumor cell aggregation. These results demonstrate that carbohydrate-mediated, metastasis associated tumor cell adhesion could be inhibited efficiently with short synthetic peptides which do not mimic naturally occurring glycoepitopes yet bind to the galectin-3 carbohydrate recognition domain with high affinity and specificity. PMID- 15528217 TI - Polymorphisms in the MMP1 and MMP3 promoter and non-small cell lung carcinoma in North China. AB - Matrix metalloproteinases (MMPs) are proteolytic enzymes that regulate various cell behaviors in cancer biology, via their basic function of degradation of proteins. Genetic variations in several MMP promoters may influence transcription and expression of MMPs. The aim of this study is to assess the effects of the two single nucleotide polymorphisms (SNPs), the guanine insertion polymorphism in the MMP1 promoter and the adenosine insertion polymorphism in the MMP3 promoter, on risk of the development and lymphatic metastasis of non-small cell lung carcinoma (NSCLC). The MMP1 and MMP3 SNPs were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis in 243 NSCLC patients and 350 control subjects in North China. The overall genotype and allelotype distribution of both the variants in cancer patients and controls was not significantly different (all P values are above 0.05). However, stratification analysis showed that smoking individuals with the MMP3 5A allele had a >1.5-fold increased risk to develop NSCLC, compared with those harboring the 6A homozygous [the age and gender adjusted odds ratio (OR) = 1.68, 95% confidence interval (CI) = 1.04-2.70]. In addition, the frequency of the MMP3 5A homozygote in NSCLC patients with lymphatic metastasis was significantly higher than that in lymph node negative ones (5.7 versus 0%, P = 0.04). Moreover, the MMP 1G/5A haplotype significantly increased the risk of lymphatic metastasis (OR = 3.36, 95% CI = 1.42-7.94), compared with the 2G/6A haplotype. The present result suggested that the MMP3 promoter polymorphism may modify susceptibility to NSCLC, and the MMP 1G/5A haplotype may predicate the risk of lymphatic metastasis of this tumor. PMID- 15528218 TI - GSTM1, GSTT1 and GSTP1 polymorphisms, environmental tobacco smoke exposure and risk of lung cancer among never smokers: a population-based study. AB - Glutathione S-transferases detoxify polycyclic aromatic hydrocarbons found in tobacco smoke by glutathione conjugation. Polymorphisms within the GSTM1, GSTT1 and GSTP1 genes, coding for enzymes with deficient or reduced activity, have been studied as potential modifiers of lung cancer risk. It is hypothesized that risk associated with potential susceptibility gene polymorphisms might be most evident at low levels of exposure. Never smokers developing lung cancer represent a highly susceptible subset of the population, exposed to tobacco carcinogens only through environmental tobacco smoke. This population-based case-control study examines the association between GSTM1, GSTT1 and GSTP1 genotypes and lung cancer in one of the largest samples of never smokers to date. Cases (n = 166) were identified through the metropolitan Detroit Surveillance, Epidemiology and End Results (SEER) program and age- and race-matched population-based controls (n = 181) were identified using random digit dialing. Overall, there was no significant association between single or combinations of genotypes at GSTM1, GSTT1 or GSTP1 and lung cancer risk after adjustment for age, race, sex and household ETS exposure in years. However, in never smokers exposed to 20 or more years of household ETS, carrying the GSTM1 null genotype was associated with a 2.3-fold increase in risk [95% confidence interval (CI) 1.05-5.13]. Individuals in this high ETS exposure category carrying the GSTM1 null and the GSTP1 Val allele were at over 4-fold increased risk of developing lung cancer (OR = 4.56, 95% CI: 1.21-17.21). These findings suggest that in the presence of ETS, the GSTM1 genotype both alone and in combination with the GSTP1 genotype alters the risk of developing lung cancer among never smokers. PMID- 15528219 TI - Skin cancer chemopreventive agent, {alpha}-santalol, induces apoptotic death of human epidermoid carcinoma A431 cells via caspase activation together with dissipation of mitochondrial membrane potential and cytochrome c release. AB - alpha-Santalol, an active component of sandalwood oil, has been studied in detail in recent years for its skin cancer preventive efficacy in murine models of skin carcinogenesis; however, the mechanism of its efficacy is not defined. Two major biological events responsible for the clonal expansion of transformed/initiated cells into tumors are uncontrolled growth and loss of apoptotic death. Accordingly, in the present study, employing human epidermoid carcinoma A431 cells, we assessed whether alpha-santalol causes cell growth inhibition and/or cell death by apoptosis. Treatment of cells with alpha-santalol at concentrations of 25-75 microM resulted in a concentration- and a time-dependent decrease in cell number, which was largely due to cell death. Fluorescence-activated cell sorting analysis of Annexin V/propidium iodide (PI) stained cells revealed that alpha-santalol induces a strong apoptosis as early as 3 h post-treatment, which increases further in a concentration- and a time-dependent manner up to 12 h. Mechanistic studies showed an involvement of caspase-3 activation and poly(ADP ribose) polymerase cleavage through activation of upstream caspase-8 and -9. Further, the treatment of cells with alpha-santalol also led to disruption of the mitochondrial membrane potential and cytochrome c release into the cytosol, thereby implicating the involvement of the mitochondrial pathway. Pre-treatment of cells with caspase-8 or -9 inhibitor, pan caspase inhibitor or cycloheximide totally blocked alpha-santalol-caused caspase-3 activity and cleavage, but only partially reversed apoptotic cell death. This suggests involvement of both caspase-dependent and -independent pathways, at least under caspase inhibiting conditions, in alpha-santalol-caused apoptosis. Together, this study for the first time identifies the apoptotic effect of alpha-santalol, and defines the mechanism of apoptotic cascade activated by this agent in A431 cells, which might be contributing to its overall cancer preventive efficacy in mouse skin cancer models. PMID- 15528220 TI - DNMT3B polymorphisms and risk of primary lung cancer. AB - DNA-methyltransferase-3B (DNMT3B) plays an important role in the generation of aberrant methylation in carcinogenesis. Polymorphisms and haplotypes of the DNMT3B gene may influence DNMT3B activity on DNA methylation, thereby modulating the susceptibility to lung cancer. To test this hypothesis, we investigated the association of the -283T > C (from exon 1A transcription start site) and -579G > T (from exon 1B transcription start site) polymorphisms in DNMT3B promoter, and their haplotypes with the risk of lung cancer in a Korean population. The DNMT3B genotype was determined in 432 lung cancer patients and 432 healthy controls that were frequency-matched for age and sex. Individuals with at least one -283T allele were at a significantly decreased risk of adenocarcinoma (AC) and small cell carcinoma (SM) [adjusted odds ratio (OR) = 0.48, 95% confidence interval (CI) = 0.28-0.82, P = 0.007; and adjusted OR = 0.47, 95% CI = 0.24-0.93, P = 0.03, respectively] compared with those harboring a -283CC genotype. Individuals with at least one -579G allele were also at a significantly decreased risk of AC and SM (adjusted OR = 0.47, 95% CI = 0.28-0.81, P = 0.006; and adjusted OR = 0.51, 95% CI = 0.26-0.99, P = 0.048, respectively) compared with those having a 579TT genotype. The -283T allele was linked with the -579G allele, and haplotype 283T/-579G was associated with a significantly decreased risk of AC (adjusted OR = 0.48, 95% CI = 0.29-0.81, P = 0.006) as compared with haplotype -283C/-579T. In a promoter assay, carriage of the -283T allele showed a significantly lower promoter activity ( approximately 50%) compared with the -283C allele (P < 0.001), but the -579G > T polymorphism did not have an affect on the DNMT3B promoter activity. These results suggest that the DNMT3B -283T > C polymorphism influences DNMT3B expression, thus contributing to the genetic susceptibility to lung cancer. PMID- 15528221 TI - Negative regulatory effect of histamine in DNFB-induced contact hypersensitivity. AB - Histamine plays an important role in the regulation of various immunological functions. To evaluate the role of histamine in contact hypersensitivity, contact dermatitis was induced with dinitrofluorobenzene (DNFB) in histidine decarboxylase knockout (HDC-/-) histamine-deficient and wild-type mice. The DNFB induced increase of the ear thickness was significantly higher in HDC-/- mice than in wild-type mice. Using flow cytometry, significantly lower percentages of CD4+ Th and CD8+ Tc cells, and significantly higher percentages of CD45R+ B cells were observed in the regional lymph nodes in HDC-/- mice than in wild-type mice. In the ear specimens of both groups, the majority of the infiltrating cells were neutrophils and macrophages at 24 and 48 h after challenge. Using immunohistochemistry, we observed significantly more CD45+ leukocytes in HDC-/- mice than in wild-type mice. The expression of Th1 (IL-2, IFN-gamma, TNF-alpha) and Th2 (IL-4) mRNAs was examined by quantitative real time RT-PCR in the ear samples. The levels of Th1 cytokine mRNAs both at 24 and 48 h after challenge and IL-4 mRNA at 48 h showed a significantly higher increase in HDC-/- mice than in wild-type mice. These results suggest that histamine plays a negative immunoregulatory role in DNFB-induced contact hypersensitivity. PMID- 15528222 TI - High dissemination and hepatotoxicity in rats infected with Candida albicans after stress exposure: potential sensitization to liver damage. AB - The liver constitutes the first barrier in the control of hematogenous dissemination for Candida albicans of intestinal origin. The ability of this organ to limit the growth of the yeast and to mount an efficient inflammatory reaction is crucial in determining the outcome of the fungal infection. When rats infected with C. albicans are exposed to chronic varied stress, the cell recruitment is impaired at the site of the infection, the tissue reaction is highly disorganized in target organs and the infection evolution is more severe. At hepatic level, higher fungal burden is associated with hyphal form and the consistent presence of steatosis (fatty liver). Herein we aimed at characterizing the steatosis associated with C. albicans infection and to provide molecular evidence of the correlation among liver injury markers, stress products and the initiation of the inflammatory tissue reaction. After 3 days of stress and infection, we observed micro and macro steatosis in acinar zone 1 (specific lipid stain), higher lipid peroxidation and increased levels of serum alanine aminotransferase and gamma glutamil transferase. While infection triggered hepatic NO production and arginase activity, stress down-modulated both. Remarkably, defects in levels of TNF-alpha and NO were observed during the first step of the inflammatory response. Our results demonstrate that stress mediators down-regulate the acute inflammatory reaction in the hepatic scenario, promoting a major liver injury with particular immunopathological traits. PMID- 15528223 TI - Linked mechanical and biological aspects of remodeling in mouse pulmonary arteries with hypoxia-induced hypertension. AB - Right heart failure due to pulmonary hypertension causes significant morbidity and mortality. To study the linked vascular mechanical and biological changes that are induced by pulmonary hypertension, we mechanically tested isolated left main pulmonary arteries from mice exposed to chronic hypobaric hypoxia and performed histological assays on contralateral vessels. In isolated vessel tests, hypoxic vessels stretched less in response to pressure than controls at all pressure levels. Given the short length and large diameter of the pulmonary artery, the tangent Young's modulus could not be measured; instead, an effective elastic modulus was calculated that increased significantly with hypoxia [(280 kPa (SD 53) and 296 kPa (SD 50) for 10 and 15 days, respectively, vs. 222 kPa (SD 35) for control; P < 0.02)]. Hypoxic vessels also had higher damping coefficients [(0.063 (SD 0.017) and 0.054 (SD 0.014) for 10 and 15 days, respectively, vs. 0.033 (SD 0.016) for control; P < 0.002)], indicating increased energy dissipation. The increased stiffness with hypoxia correlated with an increase in collagen thickness (percent collagen multiplied by wall thickness) as well as the sum of elastin and collagen thicknesses measured histologically in the artery wall. These results highlight the mechanobiological changes in the pulmonary vasculature that occur in response to hypoxia-induced pulmonary hypertension. Furthermore, they demonstrate significant vascular mechanical and biological changes that would increase pulmonary vascular impedance, leading to right heart failure. PMID- 15528224 TI - Altered muscle metaboreflex control of coronary blood flow and ventricular function in heart failure. AB - We investigated the effect of muscle metaboreflex activation on left circumflex coronary blood flow (CBF), coronary vascular conductance (CVC), and regional left ventricular performance in conscious, chronically instrumented dogs during treadmill exercise before and after the induction of heart failure (HF). In control experiments, muscle metaboreflex activation during mild exercise elicited significant reflex increases in mean arterial pressure, heart rate, and cardiac output. CBF increased significantly, whereas no significant change in CVC occurred. There was no significant change in the minimal rate of myocardial shortening (-dl/dt(min)) with muscle metaboreflex activation during mild exercise (15.5 +/- 1.3 to 16.8 +/- 2.4 mm/s, P > 0.05); however, the maximal rate of myocardial relaxation (+dl/dt(max)) increased (from 26.3 +/- 4.0 to 33.7 +/- 5.7 mm/s, P < 0.05). Similar hemodynamic responses were observed with metaboreflex activation during moderate exercise, except there were significant changes in both -dl/dt(min) and dl/dt(max). In contrast, during mild exercise with metaboreflex activation during HF, no significant increase in cardiac output occurred, despite a significant increase in heart rate, inasmuch as a significant decrease in stroke volume occurred as well. The increases in mean arterial pressure and CBF were attenuated, and a significant reduction in CVC was observed (0.74 +/- 0.14 vs. 0.62 +/- 0.12 ml x min(-1) x mmHg(-1); P < 0.05). Similar results were observed during moderate exercise in HF. Muscle metaboreflex activation did not elicit significant changes in either -dl/dt(min) or +dl/dt(max) during mild exercise in HF. We conclude that during HF the elevated muscle metaboreflex-induced increases in sympathetic tone to the heart functionally vasoconstrict the coronary vasculature, which may limit increases in myocardial performance. PMID- 15528225 TI - Nitric oxide donors protect murine myocardium against infarction via modulation of mitochondrial permeability transition. AB - Mitochondrial permeability transition (MPT) pores have recently been implicated as a potential mediator of myocardial ischemic injury. Nitric oxide (NO) donors induce a powerful late phase of cardioprotection against ischemia-reperfusion injury; however, the cellular mechanisms involved are poorly understood. The role of MPT pores as a target of cardioprotective signaling pathways activated by NO has never been explored in detail. Thus mice were administered the NO donor diethylenetriamine (DETA)/NO (4 doses of 0.1 mg/kg i.v. each) 24 h before 30 min of coronary artery occlusion followed by 24 h of reperfusion. Infarct size was significantly reduced in DETA/NO-treated mice (30 +/- 2% of risk region in treated mice vs. 50 +/- 2% in control mice; P < 0.05), which demonstrates powerful cardioprotection. To examine the role of MPT pores, mice were administered atractyloside (Atr; 25 mg/kg i.v.), which induces adenine nucleotide translocase-dependent MPT, 20 min before ischemia. Atr blocked the infarct sparing effects of DETA/NO (infarct size, 58 +/- 1 vs. 30 +/- 2% of risk region in DETA/NO; P < 0.05), whereas Atr alone had no effect. Mitochondria isolated from DETA/NO-treated mice exhibited increased resistance to Ca(2+)-induced swelling by 20 micromol/l CaCl(2) or by the higher concentration of 200 micromol/l, which suggests that cardioprotection involves decreased propensity for MPT. Preincubation of mitochondria from control hearts with 30 nmol/l of the pore inhibitor cyclosporin A prevented swelling by 200 micromol/l CaCl(2), thereby confirming that Ca(2+) induces mitochondrial swelling via MPT. In accordance with the effects on infarct size, administration of Atr to the mice significantly abrogated DETA/NO-induced protection against Ca(2+)-induced mitochondrial swelling. These phenotypic alterations were associated with an increase in the antiapoptotic protein Bcl-2, which suggests that the underlying mechanisms may involve inhibition of cell death by Bcl-2. These data suggest that a critical process during NO donor-induced cardioprotection is to prevent MPT pore opening potentially via targeting of the adenine nucleotide translocator. PMID- 15528226 TI - Functional proteomic analysis of a three-tier PKCepsilon-Akt-eNOS signaling module in cardiac protection. AB - Cardiac protective signaling networks have been shown to involve PKCepsilon. However, the molecular mechanisms by which PKCepsilon interacts with other members of these networks to form task-specific modules remain unknown. Among 93 different PKCepsilon-associated proteins that have been identified, Akt and endothelial nitric oxide (NO) synthase (eNOS) are of importance because of their independent abilities to promote cell survival and prevent cell death. The simultaneous association of PKCepsilon, Akt, and eNOS has not been examined, and, in particular, the formation of a module containing these three proteins and the role of such a module in the regulation of NO production and cardiac protection are unknown. The present study was undertaken to determine whether these molecules form a signaling module and, thereby, play a collective role in cardiac signaling. Using recombinant proteins in vitro and PKCepsilon transgenic mouse hearts, we demonstrate the following: 1) PKCepsilon, Akt, and eNOS interact and form signaling modules in vitro and in the mouse heart. Activation of either PKCepsilon or Akt enhances the formation of PKCepsilon-Akt-eNOS signaling modules. 2) PKCepsilon directly phosphorylates and enhances activation of Akt in vitro, and PKCepsilon activation increases phosphorylation and activation of Akt in PKCepsilon transgenic mouse hearts. 3) PKCepsilon directly phosphorylates eNOS in vitro, and this phosphorylation enhances eNOS activity. Activation of PKCepsilon in vivo increased phosphorylation of eNOS at Ser(1177), indicating eNOS activation. This study characterizes, for the first time, the physical, as well as functional, coupling of PKCepsilon, Akt, and eNOS in the heart and implicates these PKCepsilon-Akt-eNOS signaling modules as critical signaling elements during PKCepsilon-induced cardiac protection. PMID- 15528227 TI - Diabetic dyslipidemia and exercise affect coronary tone and differential regulation of conduit and microvessel K+ current. AB - Spontaneous transient outward K(+) currents (STOCs) elicited by Ca(2+) sparks and steady-state K(+) currents modulate vascular reactivity, but effects of artery size, diabetic dyslipidemia, and exercise on these differentially regulated K(+) currents are unclear. We studied the conduit arteries and microvessels of male Yucatan swine assigned to one of three groups for 20 wk: control (C, n = 7), diabetic dyslipidemic (DD, n = 6), or treadmill-trained DD animals (DDX, n = 7). Circumflex artery blood flow velocity obtained with intracoronary Doppler and lumen diameters obtained by intravascular ultrasound enabled calculation of absolute coronary blood flow (CBF). Ca(2+) sparks were determined in pressurized microvessels, and perforated patch clamp assessed K(+) current in smooth muscle cells isolated from conduits and microvessels. Baseline CBF in DD was decreased versus C. In pressurized microvessels, Ca(2+) spark activity was significantly lower in DD versus C and DDX (P < 0.05 vs. DDX). STOCs were pronounced in microvessel (approximately 35 STOCs/min) in sharp contrast to conduit cells ( approximately 2 STOCs/min). STOCs were decreased by 86% in DD versus C and DDX in microvessels; in contrast, there was no difference in STOCs across groups in conduit cells. Steady-state K(+) current in microvessels was decreased in DD and DDX versus C; in contrast, steady-state K(+) current in conduit cells was decreased in DDX versus DD and C. We conclude that steady-state K(+) current and STOCs are differentially regulated in conduit versus microvessels in health and diabetic dyslipidemia. Exercise prevented diabetic dyslipidemia-induced decreases in baseline CBF, possibly via STOC-regulated basal microvascular tone. PMID- 15528228 TI - Regulation of actin dynamics is critical for endothelial barrier functions. AB - We tested the hypothesis that the equilibrium between F- and G-actin in endothelial cells modulates the integrity of the actin cytoskeleton and is important for the maintenance of endothelial barrier functions in vivo and in vitro. We used the actin-depolymerizing agent cytochalasin D and jasplakinolide, an actin filament (F-actin) stabilizing and promoting substance, to modulate the actin cytoskeleton. Low doses of jasplakinolide (0.1 microM), which we have previously shown to reduce the permeability-increasing effect of cytochalasin D, had no influence on resting permeability of single-perfused mesenteric microvessels in vivo as well as on monolayer integrity. The F-actin content of cultured endothelial cells remained unchanged. In contrast, higher doses (10 microM) of jasplakinolide increased permeability (hydraulic conductivity) to the same extent as cytochalasin D and induced formation of intercellular gaps in cultured myocardial endothelial (MyEnd) cell monolayers. This was accompanied by a 34% increase of F-actin and pronounced disorganization of the actin cytoskeleton in MyEnd cells. Furthermore, we tested whether an increase of cAMP by forskolin and rolipram would prevent the cytochalasin D-induced barrier breakdown. Conditions that increase intracellular cAMP failed to block the cytochalasin D-induced permeability increase in vivo and the reduction of vascular endothelial cadherin-mediated adhesion in vitro. Taken together, these data support the hypothesis that the state of polymerization of the actin cytoskeleton is critical for maintenance of endothelial barrier functions and that both depolymerization by cytochalasin D and hyperpolymerization of actin by jasplakinolide resulted in an increase of microvessel permeability in vivo. However, cAMP, which is known to support endothelial barrier functions, seems to work by mechanisms other than stabilizing F-actin. PMID- 15528229 TI - Calpain inhibitor-1 protects the rat heart from ischemia-reperfusion injury: analysis by mechanical work and energetics. AB - We hypothesized that calpain inhibitor-1 protected left ventricular (LV) function from ischemia-reperfusion injury by inhibiting the proteolysis of alpha-fodrin. To test this hypothesis, we investigated the effect of calpain inhibitor-1 on LV mechanical work and energetics in the cross-circulated rat hearts that underwent 15-min global ischemia and 60-min reperfusion (n = 9). After ischemia-reperfusion with calpain inhibitor-1, mean end-systolic pressure at midrange LV volume and systolic pressure-volume area (PVA) at midrange LV volume (total mechanical energy per beat) were hardly changed, although they were significantly (P < 0.01) decreased after ischemia-reperfusion without calpain inhibitor-1. Mean myocardial oxygen consumption per beat (Vo(2)) intercepts (PVA-independent Vo(2); Vo(2) for the total Ca(2+) handling in excitation-contraction coupling and basal metabolism) of Vo(2)-PVA linear relations were also unchanged after ischemia reperfusion with calpain inhibitor-1, although they were significantly (P < 0.01) decreased after ischemia-reperfusion without calpain inhibitor-1. There were no significant differences in O(2) costs of LV PVA and contractility among the hearts in control (or normal) postischemia-reperfusion and postischemia reperfusion with calpain inhibitor-1. Western blot analysis of alpha-fodrin and the immunostaining of 150-kDa products of alpha-fodrin confirmed that calpain inhibitor-1 almost completely protected the proteolysis of alpha-fodrin. Our results indicate that calpain inhibitor-1 prevents the heart from ischemia reperfusion injury associated with the impairment of total Ca(2+) handling by directly inhibiting the proteolysis of alpha-fodrin. PMID- 15528230 TI - Molecular and phenotypic effects of heterozygous, homozygous, and compound heterozygote myosin heavy-chain mutations. AB - Autosomal dominant familial hypertrophic cardiomyopathy (FHC) has variable penetrance and phenotype. Heterozygous mutations in MYH7 encoding beta-myosin heavy chain are the most common causes of FHC, and we proposed that "enhanced" mutant actin-myosin function is the causative molecular abnormality. We have studied individuals from families in which members have two, one, or no mutant MYH7 alleles to examine for dose effects. In one family, a member homozygous for Lys207Gln had cardiomyopathy complicated by left ventricular dilatation, systolic impairment, atrial fibrillation, and defibrillator interventions. Only one of five heterozygous relatives had FHC. Leu908Val and Asp906Gly mutations were detected in a second family in which penetrance for Leu908Val heterozygotes was 46% (21/46) and 25% (3/12) for Asp906Gly. Despite the low penetrance, hypertrophy was severe in several heterozygotes. Two individuals with both mutations developed severe FHC. The velocities of actin translocation (V(actin)) by mutant and wild-type (WT) myosins were compared in the in vitro motility assay. Compared with WT/WT, V(actin) was 34% faster for WT/D906G and 21% for WT/L908V. Surprisingly V(actin) for Leu908Val/Asp906Gly and Lys207Gln/Lys207Gln mutants were similar to WT. The apparent enhancement of mechanical performance with mutant/WT myosin was not observed for mutant/mutant myosin. This suggests that V(actin) may be a poor predictor of disease penetrance or severity and that power production may be more appropriate, or that the limited availability of double mutant patients prohibits any definitive conclusions. Finally, severe FHC in heterozygous individuals can occur despite very low penetrance, suggesting these mutations alone are insufficient to cause FHC and that uncharacterized modifying mechanisms exert powerful influences. PMID- 15528231 TI - Effects of chronic administration of clenbuterol on function and metabolism of adult rat cardiac muscle. AB - Clenbuterol (Clen), a beta(2)-agonist, is known to produce skeletal and myocardial hypertrophy. This compound has recently been used in combination with left ventricular assist devices for the treatment of end-stage heart failure to reverse or prevent the adverse effects of unloading-induced myocardial atrophy. However, the mechanisms of action of Clen on myocardial cells have not been fully elucidated. In an attempt to clarify this issue, we examined the effects of chronic administration of Clen on Ca(2+) handling and substrate preference in cardiac muscle. Rats were treated with either 2 mg x kg(-1) x day(-1) Clen or saline (Sal) for 4 wk with the use of osmotic minipumps. Ventricular myocytes were enzymatically dissociated. Cells were field stimulated at 0.5, 1, and 2 Hz, and cytoplasmic Ca(2+) transients were monitored with the use of the fluorescent indicator indo-1 acetoxymethyl ester. Two-dimensional surface area and action potentials in current clamp were also measured. We found that in the Clen group there was significant hypertrophy at the organ and cellular levels compared with Sal. In Clen myocytes, the amplitude of the indo-1 ratio transients was significantly increased. Sarcoplasmic reticulum Ca(2+) content, estimated by rapid application of 20 mM caffeine, was significantly increased in the Clen group. The action potential was prolonged in the Clen group compared with Sal. Carbohydrate contribution to the tricarboxylic cycle (Krebs cycle) flux was increased several times in the Clen group. This increase was associated with decreased expression of peroxisome proliferator-activated receptor-alpha. This study shows that chronic administration of Clen induces cellular hypertrophy and increases oxidative carbohydrate utilization together with an increase in sarcoplasmic reticulum Ca(2+) content, which results in increased amplitude of the Ca(2+) transients. These effects could be important when Clen is used in conjunction with left ventricular assist devices treatment. PMID- 15528233 TI - Endothelium-derived 2-arachidonylglycerol: an intermediate in vasodilatory eicosanoid release in bovine coronary arteries. AB - Acetylcholine stimulates the release of endothelium-derived arachidonic acid (AA) metabolites including prostacyclin and epoxyeicosatrienoic acids (EETs), which relax coronary arteries. However, mechanisms of endothelial cell (EC) AA activation remain undefined. We propose that 2-arachidonylglycerol (2-AG) plays an important role in this pathway. An AA metabolite isolated from bovine coronary ECs was identified as 2-AG by mass spectrometry. In ECs pretreated with the fatty acid amidohydrolase inhibitor diazomethylarachidonyl ketone (DAK; 20 micromol/l), methacholine (10 micromol/l)-stimulated 2-AG release was blocked by the phospholipase C inhibitor U-73122 (10 micromol/l) or the diacylglycerol lipase inhibitor RHC-80267 (40 micromol/l). In U-46619-preconstricted bovine coronary arterial rings, 2-AG relaxations averaging 100% at 10 micromol/l were inhibited by endothelium removal, by DAK, by the hydrolase inhibitor methyl arachidonylfluorophosphate (10 micromol/l), by the cyclooxygenase inhibitor indomethacin (10 micromol/l), but not by the CB1 cannabinoid receptor antagonist SR-141716 (1 micromol/l). The cytochrome P-450 inhibitor SKF-525a (10 micromol/l) and the 14,15-epoxyeicosa-5Z-enoic acid EET antagonist (14,15-EEZE; 10 micromol/l) further attenuated the indomethacin-resistant relaxations. The nonhydrolyzable 2-AG analogs noladin ether, 2-AG amide, and 14,15-EET glycerol amide did not induce relaxation. N-nitro-L-arginine-resistant relaxations to methacholine were also inhibited by U-73122, RHC-80267, and DAK. 14,15-EET glycerol ester increased opening of large-conductance K(+) channels 12-fold in cell-attached patches of isolated smooth muscle cells and induced relaxations averaging 95%. These results suggest that methacholine stimulates EC 2-AG production through phospholipase C and diacylglycerol lipase activation. 2-AG is further hydrolyzed to AA, which is metabolized to vasoactive eicosanoids. These studies reveal a role for 2-AG in EC AA release and the regulation of coronary tone. PMID- 15528232 TI - PPAR-gamma activation fails to provide myocardial protection in ischemia and reperfusion in pigs. AB - Peroxisome proliferator-activated receptor (PPAR)-gamma modulates substrate metabolism and inflammatory responses. In experimental rats subjected to myocardial ischemia-reperfusion (I/R), thiazolidinedione PPAR-gamma activators reduce infarct size and preserve left ventricular function. Troglitazone is the only PPAR-gamma activator that has been shown to be protective in I/R in large animals. However, because troglitazone contains both alpha-tocopherol and thiazolidinedione moieties, whether PPAR-gamma activation per se is protective in myocardial I/R in large animals remains uncertain. To address this question, 56 pigs were treated orally for 8 wk with troglitazone (75 mg x kg(-1) x day(-1)), rosiglitazone (3 mg x kg(-1) x day(-1)), or alpha-tocopherol (73 mg x kg(-1) x day(-1), equimolar to troglitazone dose) or received no treatment. Pigs were then anesthetized and subjected to 90 min of low-flow regional myocardial ischemia and 90 min of reperfusion. Myocardial expression of PPAR-gamma, determined by ribonuclease protection assay, increased with troglitazone and rosiglitazone compared with no treatment. Rosiglitazone had no significant effect on myocardial contractile function (Frank-Starling relations), substrate uptake, or expression of proinflammatory cytokines during I/R compared with untreated pigs. In contrast, preservation of myocardial contractile function and lactate uptake were greater and cytokine expression was attenuated in pigs treated with troglitazone or alpha-tocopherol compared with untreated pigs. Multivariate analysis indicated that presence of an alpha-tocopherol, but not a thiazolidinedione, moiety in the test compound was significantly related to greater contractile function and lactate uptake and lower cytokine expression during I/R. We conclude that PPAR gamma activation is not protective in a porcine model of myocardial I/R. Protective effects of troglitazone are attributable to its alpha-tocopherol moiety. These findings, in conjunction with prior rat studies, suggest interspecies differences in the response to PPAR-gamma activation in the heart. PMID- 15528234 TI - Inhibition of beta-adrenergic receptor trafficking in adult cardiocytes by MAP4 decoration of microtubules. AB - Decreased beta-adrenergic receptor (beta-AR) number occurs both in animal models of cardiac hypertrophy and failure and in patients. beta-AR recycling is an important mechanism for the beta-AR resensitization that maintains a normal complement of cell surface beta-ARs. We have shown that 1) in severe pressure overload cardiac hypertrophy, there is extensive microtubule-associated protein 4 (MAP4) decoration of a dense microtubule network; and 2) MAP4 microtubule decoration inhibits muscarinic acetylcholine receptor recycling in neuroblastoma cells. We asked here whether MAP4 microtubule decoration inhibits beta-AR recycling in adult cardiocytes. [(3)H]CGP-12177 was used as a beta-AR ligand, and feline cardiocytes were isolated and infected with adenovirus containing MAP4 (AdMAP4) or beta-galactosidase (Adbeta-gal) cDNA. MAP4 decorated the microtubules extensively only in AdMAP4 cardiocytes. beta-AR agonist exposure reduced cell surface beta-AR number comparably in AdMAP4 and Adbeta-gal cardiocytes; however, after agonist withdrawal, the cell surface beta-AR number recovered to 78.4 +/- 2.9% of the pretreatment value in Adbeta-gal cardiocytes but only to 56.8 +/- 1.4% in AdMAP4 cardiocytes (P < 0.01). This result was confirmed in cardiocytes isolated from transgenic mice having cardiac-restricted MAP4 overexpression. In functional terms of cAMP generation, beta-AR agonist responsiveness of AdMAP4 cells was 47% less than that of Adbeta-gal cells. We conclude that MAP4 microtubule decoration interferes with beta-AR recycling and that this may be one mechanism for beta-AR downregulation in heart failure. PMID- 15528235 TI - Stimulation of beta-adrenoceptors inhibits store-operated channel currents via a cAMP-dependent protein kinase mechanism in rabbit portal vein myocytes. AB - Previously we have described the properties of store-operated channel currents (SOCs) in freshly dispersed rabbit portal vein smooth muscle cells. In addition to Ca(2+) store depletion these SOCs could also be activated by alpha adrenoceptor stimulation and diacylglycerol (DAG) via a protein kinase C (PKC) dependent mechanism. In the present study we have investigated the effect of beta adrenoceptor stimulation on SOCs in rabbit portal vein myocytes. With whole-cell recording the selective beta-adrenoceptor agonist isoprenaline reduced the current evoked by cyclopiazonic acid (CPA, sarcoplasmic/endoplasmic reticulum ATPase inhibitor) by over 85%. With cell-attached patch recording, bath application of isoprenaline produced a pronounced inhibition of SOC activity evoked by either CPA or the acetoxymethyl ester form of BAPTA (BAPTA-AM). SOC activity evoked by CPA, the DAG analogue, 1-oleoyl-acetyl-sn-glycerol (OAG) or the phorbol ester, phorbol-12,13-dibutyrate (PDBu) was also markedly inhibited by the adenylate cyclase activator, forskolin, and the cell-permeable non hydrolysable analogue of cyclic adenosine monophosphate (cAMP), 8-Br-cAMP. With inside-out patches, bath application of PDBu evoked channel currents with similar properties to SOCs which were inhibited by over 90% by a catalytic subunit of protein kinase A (PKA) and by 8-Br-cAMP. Moreover bath application of PKA inhibitors, H-89, KT5720 and an inhibitory peptide to quiescent cell-attached or inside-out patches, activated channel currents with similar properties to SOCs. These data suggest that in rabbit portal vein myocytes, stimulation of beta adrenoceptors inhibits SOC activity via a cAMP-dependent protein kinase signal transduction cascade. In addition it is concluded that constitutive PKA activity has a profound inhibitory effect on SOC activity in this vascular preparation. PMID- 15528236 TI - Two developmental switches in GABAergic signalling: the K+-Cl- cotransporter KCC2 and carbonic anhydrase CAVII. AB - GABAergic signalling has the unique property of 'ionic plasticity', which is based on short-term and long-term changes in the Cl(-) and HCO(3)(-) ion concentrations in the postsynaptic neurones. While short-term ionic plasticity is caused by activity-dependent, channel-mediated anion shifts, long-term ionic plasticity depends on changes in the expression patterns and kinetic regulation of molecules involved in anion homeostasis. During development the efficacy and also the qualitative nature (depolarization/excitation versus hyperpolarization/inhibition) of GABAergic transmission is influenced by the neuronal expression of two key molecules: the chloride-extruding K(+)-Cl(-) cotransporter KCC2, and the cytosolic carbonic anhydrase (CA) isoform CAVII. In rat hippocampal pyramidal neurones, a steep up-regulation of KCC2 accounts for the 'developmental switch', which converts depolarizing and excitatory GABA responses of immature neurones to classical hyperpolarizing inhibition by the end of the second postnatal week. The immature hippocampus generates large-scale network activity, which is abolished in parallel by the up-regulation of KCC2 and the consequent increase in the efficacy of neuronal Cl(-) extrusion. At around postnatal day 12 (P12), an abrupt, steep increase in intrapyramidal CAVII expression takes place, promoting excitatory responses evoked by intense GABAergic activity. This is largely caused by a GABAergic potassium transient resulting in spatially widespread neuronal depolarization and synchronous spike discharges. These facts point to CAVII as a putative target of CA inhibitors that are used as antiepileptic drugs. KCC2 expression in adult rat neurones is down regulated following epileptiform activity and/or neuronal damage by BDNF/TrkB signalling. The lifetime of membrane-associated KCC2 is very short, in the range of tens of minutes, which makes KCC2 ideally suited for mediating GABAergic ionic plasticity. In addition, factors influencing the trafficking and kinetic modulation of KCC2 as well as activation/deactivation of CAVII are obvious candidates in the ionic modulation of GABAergic responses. The down-regulation of KCC2 under pathophysiological conditions (epilepsy, damage) in mature neurones seems to reflect a 'recapitulation' of early developmental mechanisms, which may be a prerequisite for the re-establishment of connectivity in damaged brain tissue. PMID- 15528238 TI - Cholinergic modulation of hippocampal cells and circuits. AB - Septo-hippocampal cholinergic fibres ramify extensively throughout the hippocampal formation to release acetylcholine upon a diverse range of muscarinic and nicotinic acetylcholine receptors that are differentially expressed by distinct populations of neurones. The resultant modulation of cellular excitability and synaptic transmission within hippocampal circuits underlies the ability of acetylcholine to influence the dynamic properties of the hippocampal network and results in the emergence of a range of stable oscillatory network states. Recent findings suggest a multitude of actions contribute to the oscillogenic properties of acetylcholine which are principally induced by activation of muscarinic receptors but also regulated through activation of nicotinic receptor subtypes. PMID- 15528237 TI - Aspects of the homeostaic plasticity of GABAA receptor-mediated inhibition. AB - Plasticity of ligand-gated ion channels plays a critical role in nervous system development, circuit formation and refinement, and pathological processes. Recent advances have mainly focused on the plasticity of channels gated by excitatory amino acids, including their acclaimed role in learning and memory. These receptors, together with voltage-gated ion channels, have also been known to be subjected to a homeostatic form of plasticity that prevents destabilization of the neurone's function and that of the network during various physiological processes. To date, the plasticity of GABA(A) receptors has been examined mainly from a developmental and a pathological point of view. Little is known about homeostatic mechanisms governing their plasticity. This review summarizes some of the findings on the homeostatic plasticity of tonic and phasic inhibitory activity. PMID- 15528239 TI - Oscillatory activity within rat substantia gelatinosa in vitro: a role for chemical and electrical neurotransmission. AB - Although rhythmic behaviour of mammalian spinal ventral horn networks has been extensively studied little is known about oscillogenesis in the spinal dorsal horn. The aims of this in vitro study were to record and determine the underlying mechanisms of potassium-evoked network field oscillations in the substantia gelatinosa of the neonatal rat dorsal horn, a lamina involved in nociceptive processing. Transient pressure ejection of a potassium solution evoked reproducible rhythmic activity in discrete areas of the substantia gelatinosa which lasted for 5-15 s with a single prominent peak in the 4-12 Hz frequency band (7.7 +/- 0.1 Hz, n = 60). Oscillations of similar frequency and amplitude were also observed in isolated dorsal horn quadrants. Application of CNQX (10 microm) reduced peak power amplitude and integrated power area (from 4 to 12 Hz) of the power spectrum, whereas D-AP5 (50 microm) had no effect on the potassium evoked rhythm. Bicuculline (30 microm) or strychnine (10 microm) reduced the power amplitude and area. On combination of bicuculline (30 microm) and strychnine (10 microm) the reductions in power amplitude and area were not significantly different (P > 0.05) when compared with application of either drug alone. The gap junction blockers carbenoxolone (100 microm) or octanol (1 mM) significantly reduced power amplitude and area. Although TTX (1 microm) or a calcium-free perfusate both caused reductions in the power amplitude and area, potassium-evoked rhythmic activity persisted. However, this persistent rhythm was further reduced on combination of calcium-free perfusate with octanol (1 mM) and was abolished using a cocktail of drugs. Blockade of the potassium delayed rectifier current by tetraethylammonium (5 mM) or the hyperpolarization-activated current (I(h)) by ZD7288 (10 microm) disrupted the synchronization of the potassium-induced oscillation. The frequency of potassium-induced rhythms was unaffected by any of the drugs tested. These novel findings demonstrate that transient pressure ejection of potassium evokes oscillatory activity in the substantia gelatinosa in vitro. This rhythm is partly dependent upon various receptors (AMPA/kainate, GABA(A) and glycine), ion channels (potassium delayed rectifier and I(h)) and gap junctions. Oscillatory behaviour in the substantia gelatinosa could potentially play a role in the processing of nociceptive signals. PMID- 15528240 TI - Role of voltage-dependent calcium channels in stimulus-secretion coupling in rabbit carotid body chemoreceptor cells. AB - We have defined Ca2+ channel subtypes expressed in rabbit carotid body (CB) chemoreceptor cells and their participation in the stimulus-evoked catecholamine (CA) release. Ca2+ currents (I(Ca)) activated at -30 mV, peaked at +10 mV and were fully blocked by 200 microm Cd2+. L-type channels (sensitive to 2 microm nisoldipine) activated at -30 mV and carried 21 +/- 2% of total I(Ca). Non-L-type channels activated at potentials positive to -10 mV and carried: N channels (sensitive to 1 microM omega-conotoxin-GVIA) 16 +/- 1% of total I(Ca), P/Q channels (sensitive to 3 microM omega-conotoxin-MVIIC after nisoldipine plus GVIA) 23 +/- 3% of total I(Ca) and R channels (resistant to all blockers combined) 40 +/- 3% of total I(Ca). CA release induced by hypoxia, hypercapnic acidosis, dinitrophenol (DNP) and high K(+)(o) in the intact CB was inhibited by 79-98% by 200 microm Cd2+. Hypoxia, hypercapnic acidosis and DNP, depolarized chemoreceptor cells and eventually generated repetitive action potential discharge. Nisoldipine plus MVIIC nearly abolished the release of CAs induced by hypoxia and hypercapnic acidosis and reduced by 74% that induced by DNP. All these secretory responses were insensitive to GVIA. 30 and 100 mm K(+)(o) brought resting membrane potential (E(m)) of chemoreceptor cells (-48.1 +/- 1.2 mV) to 22.5 and +7.2 mV, respectively. Thirty millimolar K(+)(o)-evoked release was abolished by nisoldipine but that induced by 100 mm K(+)(o) was mediated by activation of L, N, and P/Q channels. Data show that tested stimuli depolarize rabbit CB chemoreceptor cells and elicit CA release through Ca2+ entry via voltage-activated channels. Only L and P/Q channels are tightly coupled to the secretion of CA. PMID- 15528241 TI - Frequency-dependent acceleration of relaxation in mammalian heart: a property not relying on phospholamban and SERCA2a phosphorylation. AB - An increase in stimulation frequency causes an acceleration of myocardial relaxation (FDAR). Several mechanisms have been postulated to explain this effect, among which is the Ca(2+)-calmodulin-dependent protein kinase (CaMKII) dependent phosphorylation of the Thr(17) site of phospholamban (PLN). To gain further insights into the mechanisms of FDAR, we studied the FDAR and the phosphorylation of PLN residues in perfused rat hearts, cat papillary muscles and isolated cat myocytes. This allowed us to sweep over a wide range of frequencies, in species with either positive or negative force-frequency relationships, as well as to explore the FDAR under isometric (or isovolumic) and isotonic conditions. Results were compared with those produced by isoprenaline, an intervention known to accelerate relaxation (IDAR) via PLN phosphorylation. While IDAR occurs tightly associated with a significant increase in the phosphorylation of Ser(16) and Thr(17) of PLN, FDAR occurs without significant changes in the phosphorylation of PLN residues in the intact heart and cat papillary muscles. Moreover, in intact hearts, FDAR was not associated with any significant change in the CaMKII-dependent phosphorylation of sarcoplasmic/endoplasmic Ca(2+) ATPase (SERCA2a), and was not affected by the presence of the CaMKII inhibitor, KN-93. In isolated myocytes, FDAR occurred associated with an increase in Thr(17) phosphorylation. However, for a similar relaxant effect produced by isoprenaline, the phosphorylation of PLN (Ser(16) and Thr(17)) was significantly higher in the presence of the beta-agonist. Moreover, the time course of Thr(17) phosphorylation was significantly delayed with respect to the onset of FDAR. In contrast, the time course of Ser(16) phosphorylation, the first residue that becomes phosphorylated with isoprenaline, was temporally associated with IDAR. Furthermore, KN-93 significantly decreased the phosphorylation of Thr(17) that was evoked by increasing the stimulation frequency, but failed to affect FDAR. Taken together, the results provide direct evidence indicating that CaMKII phosphorylation pathways are not involved in FDAR and that FDAR and IDAR do not share a common underlying mechanism. More likely, a CaMKII-independent mechanism could be involved, whereby increasing stimulation frequency would disrupt the SERCA2a-PLN interaction, leading to an increase in SR Ca(2+) uptake and myocardial relaxation. PMID- 15528242 TI - Layer-specific pyramidal cell oscillations evoked by tetanic stimulation in the rat hippocampal area CA1 in vitro and in vivo. AB - Tetanic stimulation of axons terminating in the CA1 region of the hippocampus induces oscillations in the gamma-to-beta frequency band (13-100 Hz) and can induce long-term potentiation (LTP). The rapid pyramidal cell discharge is driven by a mainly GABA(A)-receptor-mediated slow depolarization and entrained mainly through ephaptic interactions. This study tests whether cellular compartmentalization can explain how cells, despite severely reduced input resistance, can still fire briskly and have IPSPs superimposed on the slow GABAergic depolarization, and whether this behaviour occurs in vivo. Oscillations induced in CA1 in vitro by tetanic stimulation of the stratum radiatum or oriens were analysed using intracellular and multichannel field potentials along the cell axis. Layer-specific effects of focal application of bicuculline indicate that the GABAergic depolarization is concentrated on tetanized dendrites. Current source density analysis and characteristics of partial spikes indicate that early action potentials are initiated in the proximal nontetanized dendrite but cannot invade the tetanized dendrite, where recurrent EPSPs and evoked IPSPs were largely suppressed. As the oscillation progresses, IPSPs recover and slow the neuronal firing to beta frequencies, with a small subpopulation of neurons continuing to fire at gamma frequency. Carbonic anhydrase dependence, threshold intensity, frequency, field strength and spike initiation/propagation of tetanus evoked oscillations in urethane-anaesthetized rats, validate our observations in vitro, and show that these mechanisms operate in healthy tissue. However, the disrupted electrophysiology of the tetanized dendrites will disable normal information processing, has implications for LTP induction and is likely to play a role in pathological synchronization as found during epileptic discharges. PMID- 15528243 TI - Imaging transcription in vivo: distinct regulatory effects of fast and slow activity patterns on promoter elements from vertebrate troponin I isoform genes. AB - Firing patterns typical of slow motor units activate genes for slow isoforms of contractile proteins, but it remains unclear if there is a distinct pathway for fast isoforms or if their expression simply occurs in the absence of slow activity. Here we first show that denervation in adult soleus and EDL muscles reverses the postnatal increase in expression of troponin I (TnI) isoforms, suggesting that high-level transcription of both genes in mature muscles is under neural control. We then use a combination of in vivo transfection, live muscle imaging and fluorescence quantification to investigate the role of patterned electrical activity in the transcriptional control of troponin I slow (TnIs) and fast (TnIf) regulatory sequences by directly stimulating denervated muscles with pattern that mimic fast and slow motor units. Rat soleus muscles were electroporated with green fluorescent protein (GFP) reporter constructs harbouring 2.7 and 2.1 kb of TnIs and TnIf regulatory sequences, respectively. One week later, electrodes were implanted and muscles stimulated for 12 days. The change in GFP fluorescence of individual muscle fibres before and after the stimulation was used as a measure for transcriptional responses to different patterns of action potentials. Our results indicate that the response of TnI promoter sequences to electrical stimulation is consistent with the regulation of the endogenous genes. The TnIf and TnIs enhancers were activated by matching fast and slow activity patterns, respectively. Removal of nerve-evoked activity by denervation, or stimulation with a mismatching pattern reduced transcriptional activity of both enhancers. These results strongly suggest that distinct signalling pathways couple both fast and slow patterns of activity to enhancers that regulate transcription from the fast and slow troponin I isoforms. PMID- 15528244 TI - Effects of stretch-activated channel blockers on [Ca2+]i and muscle damage in the mdx mouse. AB - The mdx mouse lacks dystrophin and is a model of human Duchenne muscular dystrophy. Single mdx muscle fibres were isolated and subjected to a series of stretched (eccentric) contractions while measuring intracellular calcium concentration ([Ca(2+)](i)) with fluo-3 and confocal microscopy. Following the stretched contractions there was a slow rise in resting [Ca(2+)](i) and after 30 min both the [Ca(2+)](i) during a tetanus (tetanic [Ca(2+)](i)) and the tetanic force were reduced. Two blockers of stretch-activated channels, streptomycin and the spider venom toxin GsMTx4, prevented the rise of resting [Ca(2+)](i) and partially prevented the decline of tetanic [Ca(2+)](i) and force. Reducing extracellular calcium to zero also prevented the rise in resting [Ca(2+)](i) and prevented some of the decline in tetanic [Ca(2+)](i) and force. Patch-clamping experiments identified a stretch-activated channel in both wild-type and mdx myotubes which was blocked by GsMTx4. These data suggest that blockers of stretch activated channels can ameliorate the force reduction following stretched contractions by reducing the influx of Ca(2+) into the muscle. We therefore tested whether in intact mdx mice streptomycin, added to the drinking water, was capable of reducing muscle damage. mdx mice show a period of muscle damage from 20 to 40 days of life and fibres which regenerate from this damage display central nuclei. We measured the frequency of central nuclei in control mdx mice compared to streptomycin-treated mdx mice and showed that the incidence of central nuclei was significantly reduced by streptomycin treatment. This result suggests that blockers of stretch-activated channels may protect against muscle damage in the intact mdx mouse. PMID- 15528245 TI - Autogenic modulation of mechanoreceptor excitability by glutamate release from synaptic-like vesicles: evidence from the rat muscle spindle primary sensory ending. AB - Fifty-nanometre diameter, clear, synaptic-like vesicles (SLVs) are found in primary mechanosensory nerve terminals of vertebrate and invertebrate animals. We have investigated their role in mechanosensory function using the muscle spindle primary endings of rat Ia afferents as a model. Uptake and release of the synaptic vesicle marker FM1-43 indicated that SLVs recycle like synaptic vesicles and do so in a Ca(2+)-sensitive manner. Mechanical stimulation increased SLV recycling, increasing both dye uptake and release. Immunogold/electronmicroscopy showed that, like the central synaptic endings, Ia peripheral endings are enriched with glutamate. Moreover, exogenous glutamate enhanced stretch-induced Ia excitability. Enhanced excitability persisted in the presence of antagonists to the commonest ionotropic and metabotropic glutamate receptors (kynurenate, MCPG, CPPG and MAP4). However, excitation by glutamate was abolished by (R,S)-3,5 dihydroxyphenylglycine (DHPG), and rather more effectively by (2R,1'-S,2'-R,3'-S) 2-(2'-carboxy-3'-phenylcyclopropyl) glycine (PCCG-13). PCCG-13 also significantly reduced stretch-activated excitability in the absence of exogenous glutamate. These data indicate that SLVs recycle at rest, releasing glutamate, and that mechanical activity increases this process. The blockade with DHPG and PCCG-13 suggests that endogenous glutamate release acts, at least in part, through the recently described phospholipase D-linked metabotropic Glu receptor to maintain the excitability of the sensory endings. PMID- 15528246 TI - Voltage-controlled Ca2+ release and entry flux in isolated adult muscle fibres of the mouse. AB - The voltage-activated fluxes of Ca(2+) from the sarcoplasmic reticulum (SR) and from the extracellular space were studied in skeletal muscle fibres of adult mice. Single fibres of the interosseus muscle were enzymatically isolated and voltage clamped using a two-electrode technique. The fibres were perfused from the current-passing micropipette with a solution containing 15 mm EGTA and 0.2 mm of either fura-2 or the faster, lower affinity indicator fura-FF. Electrical recordings in parallel with the fluorescence measurements allowed the estimation of intramembrane gating charge movements and transmembrane Ca(2+) inward current exhibiting half-maximal activation at -7.60 +/- 1.29 and 3.0 +/- 1.44 mV, respectively. The rate of Ca(2+) release from the SR was calculated after fitting the relaxation phases of fluorescence ratio signals with a kinetic model to quantify overall Ca(2+) removal. Results obtained with the two indicators were similar. Ca(2+) release was 2-3 orders of magnitude larger than the flux carried by the L-type Ca(2+) current. At maximal depolarization (+50 mV), release flux peaked at about 3 ms after the onset of the voltage pulse and then decayed in two distinct phases. The slower phase, most likely resulting from SR depletion, indicated a decrease in lumenal Ca(2+) content by about 80% within 100 ms. Unlike in frog fibres, the kinetics of the rapid phase of decay showed no dependence on the filling state of the SR and the results provide little evidence for a substantial increase of SR permeability on depletion. The approach described here promises insight into excitation-contraction coupling in future studies of genetically altered mice. PMID- 15528247 TI - Localization and function of the Kv3.1b subunit in the rat medulla oblongata: focus on the nucleus tractus solitarii. AB - The voltage-gated potassium channel subunit Kv3.1 confers fast firing characteristics to neurones. Kv3.1b subunit immunoreactivity (Kv3.1b-IR) was widespread throughout the medulla oblongata, with labelled neurones in the gracile, cuneate and spinal trigeminal nuclei. In the nucleus of the solitary tract (NTS), Kv3.1b-IR neurones were predominantly located close to the tractus solitarius (TS) and could be GABAergic or glutamatergic. Ultrastructurally, Kv3.1b-IR was detected in NTS terminals, some of which were vagal afferents. Whole-cell current-clamp recordings from neurones near the TS revealed electrophysiological characteristics consistent with the presence of Kv3.1b subunits: short duration action potentials (4.2 +/- 1.4 ms) and high firing frequencies (68.9 +/- 5.3 Hz), both sensitive to application of TEA (0.5 mm) and 4-aminopyridine (4-AP; 30 mum). Intracellular dialysis of an anti-Kv3.1b antibody mimicked and occluded the effects of TEA and 4-AP in NTS and dorsal column nuclei neurones, but not in dorsal vagal nucleus or cerebellar Purkinje cells (which express other Kv3 subunits, but not Kv3.1b). Voltage-clamp recordings from outside-out patches from NTS neurones revealed an outward K(+) current with the basic characteristics of that carried by Kv3 channels. In NTS neurones, electrical stimulation of the TS evoked EPSPs and IPSPs, and TEA and 4-AP increased the average amplitude and decreased the paired pulse ratio, consistent with a presynaptic site of action. Synaptic inputs evoked by stimulation of a region lacking Kv3.1b-IR neurones were not affected, correlating the presence of Kv3.1b in the TS with the pharmacological effects. PMID- 15528248 TI - Recruitment of motor neuronal persistent inward currents shapes withdrawal reflexes in the frog. AB - The details of behaviour are determined by the interplay of synaptic connectivity within neuronal circuitry and the intrinsic membrane properties of individual neurones. One particularly dramatic intrinsic property displayed by neurones in many regions of the nervous system is membrane potential bistability, in which transient excitation of a neurone results in a persistent depolarization outlasting the initial excitation. Here we characterize the contribution of such intrinsic bistability, also referred to as plateau properties and mediated by persistent inward currents (PICs), in spinal motor neurones to the production of withdrawal behaviours in the frog. We performed experiments on the isolated frog spinal cord with attached hindlimb. This preparation allowed the simultaneous monitoring of muscle activations during motor behaviour and intracellular neuronal recordings. We found that PICs, following their potentiation by serotonin (5-HT), are recruited and contribute to the production of withdrawal behaviours. These properties conferred a voltage-dependent prolongation to the duration of motor neuronal activity. Consistent with this potentiation of motor neuronal PICs, 5-HT also increased the duration of evoked muscle activations. This behavioural potentiation, as well as the expression of PICs in individual neurones, was reduced following antagonism of L-type Ca(2+) channels. These results demonstrate that PICs in motor neurones can be recruited during the production of behaviour and play a role in specifying the temporal details of motor output. PMID- 15528249 TI - Neuronal mechanisms mediating the variability of somatosensory evoked potentials during sleep oscillations in cats. AB - The slow oscillation (SO) generated within the corticothalamic system is composed of active and silent states. The studies of response variability during active versus silent network states within thalamocortical system of human and animals provided inconsistent results. To investigate this inconsistency, we used electrophysiological recordings from the main structures of the somatosensory system in anaesthetized cats. Stimulation of the median nerve (MN) elicited cortical responses during all phases of SO. Cortical responses to stimulation of the medial lemniscus (ML) were virtually absent during silent periods. At the ventral-posterior lateral (VPL) level, ML stimuli elicited either EPSPs in isolation or EPSPs crowned by spikes, as a function of membrane potential. Response to MN stimuli elicited compound synaptic responses and spiked at any physiological level of membrane potential. The responses of dorsal column nuclei neurones to MN stimuli were of similar latency, but the latencies of antidromic responses to ML stimuli were variable. Thus, the variable conductance velocity of ascending prethalamic axons was the most likely cause of the barrages of synaptic events in VPL neurones mediating their firing at different level of the membrane potential. We conclude that the preserved ability of the somatosensory system to transmit the peripheral stimuli to the cerebral cortex during all the phases of sleep slow oscillation is based on the functional properties of the medial lemniscus and on the intrinsic properties of the thalamocortical cells. However the reduced firing ability of the cortical neurones during the silent state may contribute to impair sensory processing during sleep. PMID- 15528250 TI - Somato-dendritic nicotinic receptor responses recorded in vitro from the medial septal diagonal band complex of the rodent. AB - The medial septal diagonal band area (MS/DB), made up of GABAergic and cholinergic neurones, plays an essential role in the generation and modulation of the hippocampal theta rhythm. To understand the part that the cholinergic neurones might play in this activity, we sought to determine whether postsynaptic nicotinic receptor responses can be detected in slices of the rodent MS/DB by puffing on acetylcholine (ACh). Neurones were characterized electrophysiologically into GABAergic and cholinergic neurones according to previous criteria. Responses of the MS/SB neurones to ACh were various combinations of fast depolarizations (1.5-2.5 s), fast hyperpolarizations (3-4 s) and slow depolarizations (20-30 s), the latter two being blocked by atropine. The fast depolarizations were partially or not blocked with cadmium and low calcium, tetrodotoxin, and antagonists of other ionotropic receptors, and were antagonized with 25 microm mecamylamine. Pharmacological investigation of the responses showed that the alpha 7* nicotinic receptor type is associated with cholinergic neurones and 10% of the GABAergic neurones, and that non alpha 7* nicotinic receptor subtypes are associated with 50% of the GABAergic neurones. Pharmacological dissection of evoked and spontaneous postsynaptic responses, however, did not provide evidence for synaptic nicotinic receptor transmission in the MS/DB. It was concluded that nicotinic receptors, although prevalent on the somatic and/or dendritic membrane compartments of neurones in the MS/DB, are on extrasynaptic sites where they presumably play a neuromodulatory role. The presence of alpha 7* nicotinic receptors on cholinergic neurones may also render these cells specifically vulnerable to degeneration in Alzheimer's disease. PMID- 15528251 TI - Luminal bacterial flora determines physiological expression of intestinal epithelial cytoprotective heat shock proteins 25 and 72. AB - Heat shock proteins (HSP) 25 and 72 are expressed normally by surface colonocytes but not by small intestinal enterocytes. We hypothesized that luminal commensal microflora maintain the observed colonocyte HSP expression. The ability of the small intestine to respond to bacteria and their products and modulate HSPs has not been determined. The effects of luminal bacterial flora in surgically created midjejunal self-filling (SFL) vs. self-emptying (SEL) small-bowel blind loops on epithelial HSP expression were studied. HSP25 and HSP72 expression were assessed by immunoblot and immunohistochemistry. SFL were chronically colonized, whereas SEL contained levels of bacteria normal for the proximal small intestine. SFL creation significantly increased HSP25 and HSP72 expression relative to corresponding sections from SEL. Metronidazole treatment, which primarily affects anaerobic bacteria as well as a diet lacking fermentable fiber, significantly decreased SFL HSP expression. Small bowel incubation with butyrate ex vivo induced a sustained and significant upregulation of HSP25 and altered HSP72 expression, confirming the role of short-chain fatty acids. To determine whether HSPs induction altered responses to an injury, effects of the oxidant, monochloramine, on epithelial resistance and short-circuit current (I(sc)) responses to carbachol and glucose were compared. Increased SFL HSP expression was associated with protection against oxidant-induced decreases in transmural resistance and I(sc) responses to glucose, but not secretory responses to carbachol. In conclusion, luminal microflora and their metabolic byproducts direct expression of HSPs in gut epithelial cells, an effect that contributes to preservation of epithelial cell viability under conditions of stress. PMID- 15528252 TI - Epidermal growth factor reduces intestinal apoptosis in an experimental model of necrotizing enterocolitis. AB - Necrotizing enterocolitis (NEC) is a devastating intestinal disease of premature infants. Although end-stage NEC is characterized histopathologically as extensive necrosis, apoptosis may account for the initial loss of epithelium before full development of disease. We have previously shown that epidermal growth factor (EGF) reduces the incidence of NEC in a rat model. Although EGF has been shown to protect intestinal enterocytes from apoptosis, the mechanism of EGF-mediated protection against NEC is not known. The aim of this study was to investigate if EGF treatment elicits changes in expression of apoptotic markers in the ileum during the development of NEC. With the use of a well-established neonatal rat model of NEC, rats were divided into the following three experimental groups: dam fed (DF), milk formula fed (NEC), or fed with formula supplemented with 500 ng/ml EGF (NEC+EGF). Changes in ileal morphology, gene and protein expression, and histological localization of apoptotic regulators were evaluated. Anti-apoptotic Bcl-2 mRNA levels were markedly reduced and pro-apoptotic Bax mRNA levels were markedly elevated in the NEC group compared with DF controls. Supplementation of EGF into formula significantly increased anti-apoptotic Bcl-2 mRNA, whereas pro apoptotic Bax was significantly decreased. The Bax-to-Bcl-2 ratio for mRNA and protein was markedly decreased in NEC+EGF animals compared with the NEC group. The presence of caspase-3-positive epithelial cells was markedly reduced in EGF treated rats. These data suggest that alteration of the balance between pro-and anti-apoptotic proteins in the site of injury is a possible mechanism by which EGF maintains intestinal integrity and protects intestinal epithelium against NEC injury. PMID- 15528253 TI - Neuromedin B and its receptor are mitogens in both normal and malignant epithelial cells lining the colon. AB - Bombesin-like peptides are uniformly thought to act as mitogens in cancer. Yet by studying human tissues, we have recently shown that bombesin and its mammalian homologue gastrin-releasing peptide act as morphogens, promoting tumor differentiation when aberrantly upregulated in colon cancer. In contrast, little is known about the bombesin-like peptide neuromedin B (NMB) and its receptor (NMB R) in the human gastrointestinal tract. We therefore studied their presence and function in normal and malignant human colonic epithelia. Anti-NMB monoclonal antibodies were made against keyhole limpet hemocyanin (KLH)-conjugated human NMB, whereas anti-NMB-R antibodies were raised in rabbits against KLH-conjugated peptides corresponding to the third intracellular loop and COOH-terminal tail of the receptor protein. NMB antibody recognized two bands at approximately 1.2 kDa and approximately 1.5 kDa. NMB-R antibodies recognized a band at 80 kDa (predicted 43 kDa); whereas treatment with the deglycosylating agent peptide-N glycosidase generated bands at 65, 47, and 43 kDa. By immunohistochemistry, both NMB and NMB-R were expressed in normal and cancerous colonic epithelial tissues. In cancer, the amount of NMB was similar to that expressed by proliferating epithelial cells located within the crypt. In contrast, NMB-R expression was increased in cancer, with higher levels detected in better differentiated tumor cells. To assess NMB function, proliferation was determined in the nonmalignant human colonic epithelial cell line NCM-460 and in the colon cancer cell lines Caco-2 and HT-29. Exogenously added NMB was 50-100% more efficacious than gastrin releasing peptide in causing tumor cell proliferation, whereas only NMB increased NCM-460 cell proliferation. These findings indicate that NMB and its receptor are coexpressed by proliferating cells in which they act in an autocrine fashion with similar and modest potency in both normal and malignant colonic epithelial cells. PMID- 15528254 TI - Effects of the gap junction blocker glycyrrhetinic acid on gastrointestinal smooth muscle cells. AB - In the tunica muscularis of the gastrointestinal (GI) tract, gap junctions form low-resistance pathways between pacemaker cells known as interstitial cells of Cajal (ICCs) and between ICC and smooth muscle cells. Coupling via these junctions facilitates electrical slow-wave propagation and responses of smooth muscle to enteric motor nerves. Glycyrrhetinic acid (GA) has been shown to uncouple gap junctions, but previous studies have shown apparent nonspecific effects of GA in a variety of tissues. We tested the effects of GA using isometric force measurements, intracellular microelectrode recordings, the patch clamp technique, and the spread of Lucifer yellow within cultured ICC networks. In murine small intestinal muscles, beta-GA (10 muM) decreased phasic contractions and depolarized resting membrane potential. Preincubation of GA inhibited the spread of Lucifer yellow, increased input resistance, and decreased cell capacitance in ICC networks, suggesting that GA uncoupled ICCs. In patch clamp experiments of isolated jejunal myocytes, GA significantly decreased L-type Ca(2+) current in a dose-dependent manner without affecting the voltage dependence of this current. The IC(50) for Ca(2+) currents was 1.9 muM, which is lower than the concentrations used to block gap junctions. GA also significantly increased large-conductance Ca(2+)-activated K(+) currents but decreased net delayed rectifier K(+) currents, including 4-aminopyridine and tetraethylammonium resistant currents. In conclusion, the reduction of phasic contractile activity of GI muscles by GA is likely a consequence of its inhibitory effects on gap junctions and voltage-dependent Ca(2+) currents. Membrane depolarization may be a consequence of uncoupling effects of GA on gap junctions between ICCs and smooth muscles and inhibition of K(+) conductances in smooth muscle cells. PMID- 15528255 TI - Purinergic regulation of cholangiocyte secretion: identification of a novel role for P2X receptors. AB - The P2X family of ligand-gated cation channels is comprised of seven distinct isoforms activated by binding of extracellular purines. Although originally identified in neurons, there is increasing evidence for expression of P2X receptors in epithelia as well. Because ATP is released by both hepatocytes and cholangiocytes, these studies were performed to evaluate whether P2X receptors are present in cholangiocytes and contribute to local regulation of biliary secretion and bile formation. RT-PCR of cDNA from cultured normal rat cholangiocytes detected transcripts for P2X receptors 2, 3, 4, and 6; products from P2X3 and P2X4 were robust and always detectable. In cholangiocyte lysates, P2X4 protein was readily detected, and immunohistochemical staining of intact rat liver revealed P2X4 protein concentrated in intrahepatic bile ducts. To assess the functional significance of P2X4, isolated Mz-ChA-1 cells were exposed to the P2X4-preferring agonist 2',3'-O-(4-benzoyl-benzoyl)-ATP (BzATP), which activated inward currents of -18.2 + 3.0 pA/pF. In cholangiocyte monolayers, BzATP but not P2X3 agonists elicited robust Cl(-) secretory responses (short-circuit current) when applied to either the apical (DeltaI(sc) 22.1 +/- 3.3 microA) or basolateral (18.5 +/- 1.6 microA) chamber, with half-maximal stimulation at approximately 10 microM and approximately 1 microM, respectively. The response to BzATP was unaffected by suramin (not significant) and was inhibited by Cu(2+) (P < 0.01). These studies provide molecular and biochemical evidence for the presence of P2X receptors in cholangiocytes. Functional studies indicate that P2X4 is likely the primary isoform involved, representing a novel and functionally important component of the purinergic signaling complex modulating biliary secretion. PMID- 15528256 TI - Inwardly rectifying chloride channel activity in intestinal pacemaker cells. AB - Cl(-) channels are proposed to play a role in gut pacemaker activity, but little is known about the characteristics of Cl(-) channels in interstitial cells of Cajal (ICC), the intestinal pacemaker cells. The objective of the present study was to identify whole cell Cl(-) currents in ICC associated with previously observed single-channel activity and to characterize its inward rectification. Whole cell patch-clamp studies showed that ICC express an inwardly rectifying Cl( ) current that was not sensitive to changes in cation composition of the extracellular solutions. Currents were not affected by replacing all cations with N-methyl-d-glucamine (NMDG(+)). Whole cell currents followed the Cl(-) equilibrium potential and were inhibited by DIDS and 9-anthracene carboxylic acid. Ramp protocols of single-channel activity showed that inward rectification was due to reduction in single-channel open probability, not a reduction in single-channel conductance. Single-channel data led to the hypothesis that strong cooperation exists between 30-pS channels that show less cooperation at potentials positive to the reversal potential. Hence, an inwardly rectifying Cl( ) channel plays a prominent role in determining pacemaker activity in the gut. PMID- 15528257 TI - Fat absorption in cystic fibrosis mice is impeded by defective lipolysis and post lipolytic events. AB - Cystic fibrosis (CF) is frequently associated with progressive loss of exocrine pancreas function, leading to incomplete digestion and absorption of dietary fat. Supplementing patients with pancreatic lipase reduces fat excretion, but it does not completely correct fat malabsorption, indicating that additional pathological processes affect lipolysis and/or uptake of lipolytic products. To delineate the role of such (post) lipolytic processes in CF-related fat malabsorption, we assessed fat absorption, lipolysis, and fatty acid uptake in two murine CF models by measuring fecal fat excretion and uptake of oleate- and triolein-derived lipid. Pancreatic and biliary function was investigated by determining lipase secretion and biliary bile salt (BS) secretion, respectively. A marked increase in fecal fat excretion was observed in cftr null mice but not in homozygous DeltaF508 mice. Fecal BS loss was enhanced in both CF models, but biliary BS secretion rates were similar. Uptake of free fatty acid was delayed in both CF models, but only in null mice was a specific reduction in lipolytic activity apparent, characterized by strongly reduced triglyceride absorption. Impaired lipolysis was not due to reduced pancreatic lipase secretion. Suppression of gastric acid secretion partially restored lipolytic activity and lipid uptake, indicating that incomplete neutralization of gastric acid impedes fat absorption. We conclude that fat malabsorption in cftr null mice is caused by impairment of lipolysis, which may result from aberrant duodenal pH regulation. PMID- 15528258 TI - Interleukin-4- and -13-induced hypercontractility of human intestinal muscle cells-implication for motility changes in Crohn's disease. AB - Crohn's disease is an idiopathic inflammatory condition. However, little is known about the changes that occur in the muscularis externa, despite the fact that this tissue contributes to motility changes and stricture formation. We characterized immune activity in the muscularis externa from intestinal segments of Crohn's disease patients and evaluated the role of IL-4 and -13 as well as signal transducer and activator of transcription (STAT)6 in the contractility of the cultured human intestinal smooth muscle cells. CD3+ve cells (P < 0.01) and IL 4 protein (P < 0.01) were significantly increased in the muscularis externa of Crohn's disease patients compared with noninflamed controls. Preincubation of human cultured smooth muscle cells with IL-4 (P < 0.001) or IL-13 (P < 0.05) significantly enhanced carbachol-induced contraction, and this was significantly inhibited by the STAT6 inhibitor leflunomide (P < 0.0001). A similar profile was observed in muscle cells isolated from Crohn's disease patients. Both IL-4 and IL 13 increased specific STAT6-DNA binding in control cells, and this was inhibited by anti-STAT6 Ab (P < 0.05) or leflunomide (P < 0.05). IL-4 and IL-13 mediate the hypercontractility of intestinal muscle via a STAT6 pathway at the level of the smooth muscle cell. The STAT6 pathway may contribute to the hypercontractility of intestinal muscle in Crohn's disease. PMID- 15528259 TI - Increased protein level of PEPT1 intestinal H+-peptide cotransporter upregulates absorption of glycylsarcosine and ceftibuten in 5/6 nephrectomized rats. AB - In chronic renal failure (CRF), dietary protein is one of the factors that deteriorates residual renal functions. Numerous studies have indicated that the products of protein digestion are mainly absorbed as small peptides. However, how small peptides are absorbed in CRF remains poorly understood. H(+)-coupled peptide transporter (PEPT1/SLC15A1) plays an important role in the absorption of small peptides and peptide-like drugs in the small intestine. Because dietary protein intake is one of the risk factors for renal failure, the alteration of intestinal PEPT1 might have implications in the progression of renal disease as well as the pharmacokinetics of peptide-like drugs. In this study, we examined the alteration of intestinal PEPT1 in 5/6 nephrectomized (5/6 NR) rats, extensively used as a model of chronic renal failure. Absorption of [(14)C]glycylsarcosine and ceftibuten was significantly increased in 5/6 NR rats compared with sham-operated rats, without a change in intestinal protease activity. Western blot analysis indicated that the amount of intestinal PEPT1 protein in 5/6 NR rats was increased mainly at the upper region. On the other hand, the amount of intestinal PEPT1 mRNA was not significantly different from that of sham-operated rats. These findings indicate that the increase in absorption of small peptides and peptide-like drugs, caused by the upregulation of intestinal PEPT1 protein, might contribute to the progression of renal failure as well as the alteration of drug pharmacokinetics. PMID- 15528260 TI - Relapsing-remitting multiple sclerosis: metabolic abnormality in nonenhancing lesions and normal-appearing white matter at MR imaging: initial experience. AB - PURPOSE: To quantify, with three-dimensional proton magnetic resonance (MR) spectroscopy, metabolic characteristics of normal-appearing white matter and nonenhancing lesions in patients with relapsing-remitting multiple sclerosis (MS). MATERIALS AND METHODS: Institutional review board approval and informed patient consent were obtained. Nine patients with relapsing-remitting MS (six women, three men) and nine age-matched control subjects (seven women, two men) were studied with T1- and T2-weighted MR imaging and three-dimensional proton MR spectroscopy at spatial resolution less than a cubic centimeter. Absolute N acetylaspartate (NAA), creatine (Cr), and choline (Cho) levels were obtained from 171 voxels: 66 from lesions on T2-weighted MR images (43 hypointense and 23 isointense on T1-weighted MR images), 31 from normal-appearing white matter, and 74 from analogous normal white matter regions on images in control subjects. RESULTS: Mean NAA level in hypointense lesions (5.30 mmol/L +/- 2.27 [standard deviation]) was significantly lower (P < or = .05) than that in isointense lesions (7.82 mmol/L +/- 2.28), normal-appearing white matter (7.37 mmol/L +/- 1.71), and normal white matter in control subjects (8.89 mmol/L +/- 1.54). Cho (1.79 mmol/L +/- 0.65) and Cr (5.64 mmol/L +/- 1.50) levels in isointense lesions were indistinguishable from those in normal-appearing white matter (1.74 mmol/L +/- 0.46 and 4.99 mmol/L +/- 0.97, respectively) but were significantly higher (Cho, 20%; Cr, 24%) than those in normal white matter in control subjects (1.44 mmol/L +/- 0.40 and 4.30 mmol/L +/- 1.32, respectively). NAA, Cho, and Cr levels in normal-appearing white matter were significantly different than those in normal white matter in control subjects (NAA, 20% lower; Cho, 14% higher; and Cr, 17% higher). CONCLUSION: Abnormal metabolic activity persists in all MS tissue types. Increased Cr and Cho levels suggest (a) ongoing gliosis and attempted remyelination in isointense lesions on T1-weighted MR images and (b) membrane turnover (de- and remyelination), in addition to increased cellularity (gliosis, inflammation) in normal-appearing white matter. PMID- 15528261 TI - Training, competency, and credentialing standards for diagnostic cervicocerebral angiography, carotid stenting, and cerebrovascular intervention: a joint statement from the American Academy of Neurology, American Association of Neurological Surgeons, American Society of Interventional and Therapeutic Radiology, American Society of Neuroradiology, Congress of Neurological Surgeons, AANS/CNS Cerebrovascular Section, and Society of Interventional Radiology. PMID- 15528262 TI - Aseptic technology of vitrification of human pronuclear oocytes using open-pulled straws. AB - BACKGROUND: The aim of this study was to compare the viability of human pronuclear oocytes subjected to vitrification using cooling by direct submerging of open-pulled straws in liquid nitrogen versus vitrification by cooling of open pulled straws located inside a closed 0.5 ml straw (aseptic system). METHODS: Two and three-pronuclei stage oocytes (n=114) were cryopreserved in super-open pulled straws by vitrification in 20% ethylene glycol +20% dimethylsulphoxide (DMSO) + osmotic active and neutral non-permeable cryoprotectants with a four step exposure in 20, 33, 50 and 100% vitrification solution for 2, 1 and 1 min, and 30-50 s, respectively at room temperature, and plunging into liquid nitrogen. Oocytes of group 1 (n=42) were rapidly cooled at a speed of 20,000 degrees C/min by direct plunging of open-pulled straws into liquid nitrogen. Oocytes of group 2 (n=44) were first located in 0.5 ml straws, which were closed at both sides by metal balls, and then plunged into liquid nitrogen. This method resulted in a cooling speed of 200 degrees C/min. For both groups, oocytes were thawed rapidly at a speed of 20 000 degrees C/min using an identical protocol. Oocytes subsequently were expelled into a graded series of sucrose solutions (1.0, 0.75, 0.5, 0.25 and 0.12 mol/l) at 2.5 min intervals. RESULTS: Oocyte development up to expanded blastocyst stage after in vitro culture was 15% in group 1, 14% in group 2 and 29% in an untreated control group. CONCLUSION: The deposition of human pronuclear oocytes in open-pulled straws which are placed inside a hermetically closed container guarantees a complete isolation of oocytes from liquid nitrogen and avoids potential contamination by pathogenic microorganisms. The combination of direct plunging of this container into liquid nitrogen and rapid warming makes this process as efficient as conventional vitrification. PMID- 15528263 TI - In vitro oocyte maturation for the treatment of infertility associated with polycystic ovarian syndrome: the French experience. AB - BACKGROUND: In vitro oocyte maturation (IVM) permits the use of immature oocytes in IVF. IVM does not require ovarian stimulation and so can be offered to patients at risk of ovarian hyperstimulation syndrome. METHODS: For this indication, we carried out 45 cycles of IVM in 33 women with polycystic ovarian syndrome (PCOS). RESULTS: A total of 509 cumulus-oocyte complexes was obtained; 276 (54.2%) oocytes matured in 24 h and 45 (8.8%) in 48 h. The normal fertilization (2PN) rate of oocytes matured in 24 and 48 h was 69.5 and 73.3% respectively. Among the 214 embryos obtained, 103 were transferred and 30 were frozen. Forty transfers were performed (2.5 embryos/transfer). Eleven women had a positive beta-hCG test (26.2% of pregnancies/puncture, 27.5% of pregnancies/transfer) and nine women had a clinical pregnancy (20.0% of pregnancies/puncture, 22.5% of pregnancies/transfer). Five babies have been born and one pregnancy is ongoing. Results of the clinical examination carried out at birth were normal. CONCLUSIONS: Our results show that IVM may be offered as an alternative to conventional IVF and to ovarian drilling in women with PCOS. The role of IVM in the therapeutic armamentarium for this condition should be further clarified. PMID- 15528264 TI - Mitotic and meiotic behaviour of a naturally transmitted ring Y chromosome: reproductive risk evaluation. AB - BACKGROUND: The mitotic and meiotic behaviour of a transmitted ring Y [r(Y)] chromosome from a father to his Klinefelter syndrome (KS) son, and the mechanism of ring formation are analysed herein. To our knowledge, this is the first reported case of natural transmission of an r(Y). METHODS AND RESULTS: Amplification of X chromosome polymorphisms by PCR showed that the KS was of paternal origin. G-banding and fluorescence in situ hybridization (FISH) studies revealed a similar percentage of mosaicism in father and son by mitotic loss of r(Y). SRY gene and Y marker amplification by PCR, FISH with subtelomeric probes for Xp/Yp and Xq/Yq, and comparative genomic hybridization (CGH) analyses indicated the intactness of the Y chromosome from SRY to subtelomere Yq. FISH analysis of sperm from the father showed significantly higher frequencies (P<0.005) for diploidy and for 6, 13, 18, 21, 22, XX, XY disomies than those observed in control donors. CONCLUSIONS: An r(Y) with low material loss can be naturally transmitted, showing similar mitotic behaviour in the offspring. The presence of an r(Y) chromosome in germinal cells increased the risk of fathering offspring with numerical abnormalities, even for chromosomes not involved in the arrangement. PMID- 15528265 TI - Bilateral oophorectomy in a pregnant woman: hormonal profile from late gestation to post-partum: case report. AB - BACKGROUND: A 16 week pregnant woman presented with massive theca-lutein cysts requiring bilateral oophorectomy. Pregnancy progressed uneventfully and spontaneous lactation ensued after delivery. METHODS: To study the role of the ovary on the hormonal profile at the end of gestation and in post-partum, we measured FSH, estradiol (E2), unconjugated estrone (E1), unconjugated estriol (E3), sex hormone-binding globulin, progesterone, dehydroepiandrosterone sulphate and prolactin at 37 weeks gestation and at 8 h, 4 days, 5 weeks, and 2 months post-partum. RESULTS: These hormones were within the range expected for ovary intact pregnant and puerperal women until 4 days post-partum. At 5 weeks post partum, FSH increased to a peri-menopausal range (31.4 IU/l) while estrogens remained within the normal puerperal range (E2=239 pmol/l; E1=102 pmol/l), contrasting with their rapid changes in non-pregnant women after bilateral oophorectomy. At 2 months, while partially breastfeeding, FSH, E2 and E1 were closer to menopausal range (68 IU/l, 136 and 70.2 pmol/l respectively), and hormone replacement was started. CONCLUSIONS: We conclude that the ovary is not required to maintain a normal hormonal profile in late pregnancy and early puerperium. However, the increase in FSH to peri-menopausal levels at 5 weeks post-partum, despite breastfeeding, suggests that the ovary is needed to maintain low FSH concentrations during lactation. PMID- 15528266 TI - Glucose transporter isoform 4 is expressed in the syncytiotrophoblast of first trimester human placenta. AB - BACKGROUND: Placental glucose transport mechanisms in early pregnancy are poorly understood. The aims of this study were to investigate the expression of glucose transporter (GLUT) isoforms 1, 3 and 4 in first trimester villous tissue, to assess the effects of insulin on glucose uptake and compare them with term. METHODS: The expression of GLUT isoforms was investigated using immunohistochemistry, Western blot and reverse transcription (RT)-PCR in trophoblast tissue from terminations at 6-13 weeks gestation and term. The effects of insulin (300 ng/ml, 1 h) on glucose uptake were studied in villous fragments. RESULTS: In the first trimester, GLUT1 and GLUT3 were present in the microvillous membrane and the cytotrophoblast, and GLUT4 in perinuclear membranes in the cytosol of the syncytiotrophoblast (ST). GLUT4 protein (48 kDa) and mRNA were identified in trophoblast homogenates. Whereas GLUT1 was expressed abundantly in term placenta, the expression of GLUT3 and 4 was markedly lower at term compared with first trimester. Insulin increased glucose uptake by 182% (n=6, P<0.05) in first trimester fragments, but not in term fragments. CONCLUSIONS: The insulin-regulatable GLUT4 is expressed in the cytosol of first trimester ST compatible with a role for GLUT4 in placental glucose transport in early pregnancy. The placental expression pattern of GLUT isoforms in early pregnancy is distinct from that later in pregnancy. PMID- 15528267 TI - Proteomic analysis identifies immunophilin FK506 binding protein 4 (FKBP52) as a downstream target of Hoxa10 in the periimplantation mouse uterus. AB - The process of implantation absolutely requires synchronized development of the blastocyst to implantation competency, differentiation of the uterus to the receptive state, and a reciprocal dialogue between the blastocyst and uterine luminal epithelium. Genetic and molecular approaches have identified several signaling pathways that are critical to this process. The transcription factor Hoxa10 is one such critical player in implantation. Hoxa10-/- female mice have implantation and decidualization failure due specifically to reduced uterine responsiveness to progesterone and defective stromal cell proliferation during uterine receptivity and implantation. However, the downstream signaling pathways of Hoxa10 in these events remain largely unknown. Using the proteomics approach of difference gel electrophoresis, we have identified an immunophilin FKBP52 (FK506 binding protein 4) as one of the Hoxa10-mediated signaling molecules in the uterus. We found that FKBP52, a cochaperone protein known to influence steroid hormone receptor functions, is down-regulated in stromal cells of Hoxa10 /- mice. More importantly, FKBP52 shows differential uterine cell-specific expression during the periimplantation period. Whereas it is primarily expressed in the uterine epithelium on d 1 of pregnancy, the expression expands to the stroma on d 4 during the period of uterine receptivity and becomes localized to decidualizing stromal cells surrounding the implantation site on d 5. This suggests that FKBP52 is important for the attainment of uterine receptivity and implantation. Furthermore, FKBP52 shows differential cell-specific expression in the uterus in response to progesterone and/or estrogen consistent with its expression patterns during the periimplantation period. Collectively, these results and the female infertility phenotype of FKBP52 suggest that a Hoxa10 FKBP52 signaling axis is critical to uterine receptivity and implantation. PMID- 15528268 TI - Characterization of glucagon-like peptide-1 receptor beta-arrestin 2 interaction: a high-affinity receptor phenotype. AB - To dissect the interaction between beta-arrestin ((beta)arr) and family B G protein-coupled receptors, we constructed fusion proteins between the glucagon like peptide 1 receptor and (beta)arr2. The fusion constructs had an increase in apparent affinity selectively for glucagon, suggesting that (beta)arr2 interaction locks the receptor in a high-affinity conformation, which can be explored by some, but not all, ligands. The fusion constructs adopted a signaling phenotype governed by the tethered (beta)arr2 with an attenuated G protein mediated cAMP signal and a higher maximal internalization compared with wild-type receptors. This distinct phenotype of the fusion proteins can not be mimicked by coexpressing wild-type receptor with (beta)arr2. However, when the wild-type receptor was coexpressed with both (beta)arr2 and G protein-coupled receptor kinase 5, a phenotype similar to that observed for the fusion constructs was observed. We conclude that the glucagon-like peptide 1 fusion construct mimics the natural interaction of the receptor with (beta)arr2 with respect to binding peptide ligands, G protein-mediated signaling and internalization, and that this distinct molecular phenotype is reminiscent of that which has previously been characterized for family A G protein-coupled receptors, suggesting similarities in the effect of (beta)arr interaction between family A and B receptors also at the molecular level. PMID- 15528269 TI - Characterization of the cholesterol recognition amino acid consensus sequence of the peripheral-type benzodiazepine receptor. AB - We previously defined a cholesterol recognition/interaction amino acid consensus sequence [CRAC: L/V-X (1-5)-Y-X (1-5)-R/K] in the carboxyl terminus of the peripheral-type benzodiazepine receptor (PBR), a high-affinity drug and cholesterol-binding protein present in the outer mitochondrial membrane protein. This protein is involved in the regulation of cholesterol transport into the mitochondria, the rate-determining step in steroid biosynthesis. Reconstituted wild-type recombinant PBR into proteoliposomes demonstrated high-affinity 2 chlorophenyl)-N-methyl-N-(1-methyl-propyl)-3-isoquinolinecarboxamide and cholesterol binding. In the present work, we functionally and structurally characterized this CRAC motif using reconstituted recombinant PBR and nuclear magnetic resonance. Deletion of the C-terminal domain of PBR and mutation of the highly conserved among all PBR amino acid sequences Y152 of the CRAC domain resulted in loss of the ability of mutant recPBR to bind cholesterol. Nuclear magnetic resonance analysis of a PBR C-terminal peptide (144-169) containing the CRAC domain indicated a helical conformation for the L144-S159 fragment. As a result of the side-chain distribution, a groove that could fit a cholesterol molecule is delineated, on one hand, by Y152, T148, and L144, and, on the other hand, by Y153, M149, and A145. The aromatic rings of Y152 and Y153 assigned as essential residues for cholesterol binding constitute the gate of the groove. Furthermore, the side chain of R156 may cap the groove by interacting with the sterol hydroxyl group. These results provide structural and functional evidence supporting the finding that the CRAC domain in the cytosolic carboxyl-terminal domain of PBR might be responsible for the uptake and translocation of cholesterol into the mitochondria. PMID- 15528270 TI - The Src kinase pathway promotes tamoxifen agonist action in Ishikawa endometrial cells through phosphorylation-dependent stabilization of estrogen receptor (alpha) promoter interaction and elevated steroid receptor coactivator 1 activity. AB - Tamoxifen is the most widely used selective estrogen receptor modulator for breast cancer in clinical use today. However, tamoxifen agonist action in endometrium remains a major hurdle for tamoxifen therapy. Activation of the nonreceptor tyrosine kinase src promotes tamoxifen agonist action, although the mechanisms remain unclear. To examine these mechanisms, the effect of src kinase on estrogen and tamoxifen signaling in tamoxifen-resistant Ishikawa endometrial adenocarcinoma cells was assessed. A novel connection was identified between src kinase and serine 167 phosphorylation in estrogen receptor (ER)-alpha via activation of AKT kinase. Serine 167 phosphorylation stabilized ER interaction with endogenous ER-dependent promoters. Src kinase exhibited the additional function of potentiating the transcriptional activity of Gal-steroid receptor coactivator 1 (SRC-1) and Gal-cAMP response element binding protein-binding protein in endometrial cancer cells while having no effect on Gal-p300-associated factor and Gal fusions of the other p160 coactivators glucocorticoid-interacting protein 1 (transcriptional intermediary factor 2/nuclear coactivator-2/SRC-2) and amplified in breast cancer 1 (receptor-associated coactivator 3/activator of transcription of nuclear receptor/SRC-3). Src effects on ER phosphorylation and SRC-1 activity both contributed to tamoxifen agonist action on ER-dependent gene expression in Ishikawa cells. Taken together, these data demonstrate that src kinase potentiates tamoxifen agonist action through serine 167-dependent stabilization of ER promoter interaction and through elevation of SRC-1 and cAMP response element binding protein-binding protein coactivation of ER. PMID- 15528271 TI - Identification of target genes involved in the antiproliferative effect of glucocorticoids reveals a role for nuclear factor-(kappa)B repression. AB - Glucocorticoid hormones (GCs) exert an antiproliferative effect on most cells. However, the molecular mechanism is still largely unclear. We investigated the antiproliferative mechanism by GCs in human embryonic kidney 293 cells with stably introduced glucocorticoid receptor (GR) mutants that discriminate between cross-talk with nuclear factor-(kappa)B (NF-(kappa)B) and activator protein-1 signaling, transactivation and transrepression, and antiproliferative vs. non antiproliferative responses. Using the GR mutants, we here demonstrate a correlation between repression of NF-(kappa)B signaling and antiproliferative response. Gene expression profiling of endogenous genes in cells containing mutant GRs identified a limited number of genes that correlated with the antiproliferative response. This included a GC-mediated up-regulation of the NF (kappa)B-inhibitory protein I(kappa)B(alpha), in line with repression of NF (kappa)B signaling being important in the GC-mediated antiproliferative response. Interestingly, the GC-stimulated expression of I(kappa)B(alpha) was a direct effect despite the inability of the GR mutant to transactivate through a GC responsive element. Selective expression of I(kappa)B(alpha) in human embryonic kidney 293 cells resulted in a decreased percentage of cells in the S/G2/M phase and impaired cell proliferation. These results demonstrate that GC-mediated inhibition of NF-(kappa)B is an important mechanism in the antiproliferative response to GCs. PMID- 15528272 TI - Ligand- and cell-specific effects of signal transduction pathway inhibitors on progestin-induced vascular endothelial growth factor levels in human breast cancer cells. AB - We evaluated the signaling pathways involved in regulating vascular endothelial growth factor (VEGF), a potent angiogenic growth factor, in response to natural and synthetic progestins in breast cancer cells. Inhibition of the phosphoinositide-3'-kinase (PI3-kinase) signaling pathway or the specificity protein-1 (SP-1) transcription factor abolished both progesterone- and medroxyprogesterone acetate (MPA)-induced VEGF secretion from BT-474 and T47 DCO)cells. Inhibitors of the MAPK kinase 1/2/MAPK and N-terminal jun kinase/MAPK signaling pathways blocked both progesterone- and MPA-induced VEGF secretion in BT-474 cells. However, these inhibitors blocked only progesterone-, but not MPA induced VEGF secretion in T47-DCO cells. Inhibitors of PI3-kinase or SP-1 blocked both progesterone- and MPA-induced increases in VEGF mRNA levels in T47-DCO cells. The proximal SP-1 sites within the VEGF promoter were critical for progestin-dependent induction of VEGF. In contrast, MAPK inhibitors did not block the progesterone- or MPA-induced increases in VEGF mRNA in T47-DCO cells, suggesting that MAPK inhibitors decreased progesterone-induced VEGF secretion in T47-DCO cells by blocking posttranscriptional mechanisms. The MAPK kinase/ERK/MAPK-independent induction of VEGF mediated by MPA was associated with the PRB [progesterone receptor (PR) B] isoform of the PR in T47-DCO cells. None of the inhibitors tested reduced basal PR levels or abrogated PR-dependent gene expression from a reporter plasmid, indicating that loss of PR function cannot explain any of the observed effects. Because the PI3-kinase signaling pathway and SP-1 transcription factor play critical roles in progestin-dependent VEGF induction, these may be useful targets for developing antiangiogenic therapies to prevent progression of progestin-dependent human breast cancers. PMID- 15528273 TI - Global uterine genomics in vivo: microarray evaluation of the estrogen receptor alpha-growth factor cross-talk mechanism. AB - Cross-talk between growth factor receptors and the estrogen receptor (ER) has been proposed as a signaling mechanism in estrogen target tissues, with ER(alpha) as a direct target of growth factor receptor-activated signals, leading to regulation of estrogen target genes and estrogen-like biological responses to growth factors. We evaluated whether global genomic changes in the mouse uterus in response to epidermal growth factor or IGF-I mimic those of estradiol (E2), reflecting the cross-talk mechanism. Overlapping responses to growth factors and E2 were expected in the wild type (WT) whereas no response was expected in mice lacking ER(alpha) (ER(alpha) knockout). Surprisingly, although most of the E2 response in the WT also occurred after growth factor treatment, some genes were induced only by E2. Second, although E2 did not induce gene changes in the ER(alpha) knockout, the growth factor response was almost indistinguishable from that of the WT. Differences in response of some genes to IGF-I or epidermal growth factor indicated selective regulation mechanisms, such as phosphatidylinositol 3-kinase or MAPK-dependent responses. The robust ER(alpha) independent genomic response to growth factor observed here is surprising considering that the biological growth response is ER(alpha) dependent. We propose two mechanisms as alternatives to the cross-talk mechanism for uterine gene regulation. First, E2 increases uterine growth factors, which activate downstream signaling cascades, resulting in gene regulation. Second, growth factors and estrogen regulate similar genes. Our results suggest that the estrogen response in the uterus involves E2-specific ER(alpha)-mediated responses as well as responses resulting from convergence of growth factor and ER-initiated activities. PMID- 15528274 TI - Interaction between insulin-like growth factor-I receptor and alphaVbeta3 integrin linked signaling pathways: cellular responses to changes in multiple signaling inputs. AB - Integrins are heterodimeric transmembrane proteins that mediate cell attachment to extracellular matrix, migration, division, and inhibition of apoptosis. Because growth factors are also important for these processes, there has been interest in cooperative signaling between growth factor receptors and integrins. IGF-I is an important growth factor for vascular cells. One integrin, alphaVbeta3, that is expressed in smooth muscle cells modulates IGF-I actions. Ligand occupancy of alphaVbeta3 is required for IGF-I to stimulate cell migration and division. Src homology 2 containing tyrosine phosphatase (SHP-2) is a tyrosine phosphatase whose recruitment to signaling molecules is stimulated by growth factors including IGF-I. If alphaVbeta3 ligand occupancy is inhibited, there is no recruitment of SHP-2 to alphaVbeta3 and its transfer to downstream signaling molecules is blocked. Ligand occupancy of alphaVbeta3 stimulates tyrosine phosphorylation of the beta3-subunit, resulting in recruitment of SHP-2. This transfer is mediated by an insulin receptor substrate-1-related protein termed DOK-1. Subsequently, SHP-2 is transferred to another transmembrane protein, SHPS-1. This transfer requires IGF-I receptor-mediated tyrosine phosphorylation of SHPS-1, which contains two YXXL motifs that mediate SHP-2 binding. The transfer of SHP-2 to SHPS-1 is also required for recruitment of Shc to SHPS-1. Ligand occupancy of alphaVbeta3 results in sustained Shc phosphorylation and enhanced Shc recruitment. Shc activation results in induction of MAPK. Inhibition of the Shc/SHPS-1 complex formation results in failure to achieve sustained MAPK activation and an attenuated mitogenic response. Thus, within the vessel wall, a mechanism exists whereby ligand occupancy of the alphaVbeta3 integrin is required for assembly of a multicomponent membrane signaling complex that is necessary for cells to respond optimally to IGF-I. PMID- 15528275 TI - Identification of a functional vitamin D response element in the murine Insig-2 promoter and its potential role in the differentiation of 3T3-L1 preadipocytes. AB - Insulin-induced gene-1 (Insig-1) and its homolog Insig-2 encode closely related proteins of the endoplasmic reticulum that block proteolytic activation of sterol regulatory element binding proteins, membrane-bound transcription factors that activate synthesis of cholesterol and fatty acids in animal cells. These proteins also restrict lipogenesis in mature adipocytes and block differentiation of preadipocytes. Herein, we identified a novel 1alpha,25-dihydroxyvitamin D3 [1,25 (OH)2D3] response element in the promoter region of Insig-2 gene, which specifically binds to the heterodimer of retinoid X receptor and vitamin D receptor (VDR) and directs VDR-mediated transcriptional activation in a 1,25 (OH)2D3-dependent manner. Interestingly, 1,25-(OH)2D3 is known to directly suppress the expression of peroxisome proliferator-activated receptor gamma2 protein and inhibits adipocyte differentiation of 3T3-L1 preadipocytes and murine bone marrow stromal cells. Consistent with an idea that the antiadipogenic action of 1,25-(OH)2D3 may also involve up-regulation of Insig-2, we found that 1,25 (OH)2D3 transiently but strongly induces Insig-2 expression in 3T3-L1 cells. This novel regulatory circuit may also play important roles in other lipogenic cell types that express VDR, and collectively our results suggest an intriguing, new linkage between 1,25-(OH)2D3 and lipogenesis. PMID- 15528276 TI - A screen for genes that suppress loss of contact inhibition: identification of ING4 as a candidate tumor suppressor gene in human cancer. AB - We have devised a screen for genes that suppress the loss of contact inhibition elicited by overexpression of the protooncogene MYCN. The initial application of this screen detected nine distinctive suppressors within a representative human cDNA library. One of these genes was ING4, a potential tumor suppressor gene that maps to human chromosome 12p13. Ectopic expression of ING4 suppressed the loss of contact inhibition elicited by either MYCN or MYC but had no direct effect on cellular proliferation. Pursuing the possibility that ING4 might be a tumor suppressor gene, we found inactivating mutations in ING4 transcripts from various human cancer cell lines. In addition, we used comparative genomic hybridization to detect deletion of the ING4 locus in 10-20% of human breast cancer cell lines and primary breast tumors. Ectopic expression of ING4 attenuated the growth of T47D human breast cancer cells in soft agar. We conclude that ING4 is a strong candidate as a tumor suppressor gene. PMID- 15528277 TI - Crystal structures of 2-acetylaminofluorene and 2-aminofluorene in complex with T7 DNA polymerase reveal mechanisms of mutagenesis. AB - The carcinogen 2-acetylaminofluorene forms two major DNA adducts: N-(2' deoxyguanosin-8-yl)-2-acetylaminofluorene (dG-AAF) and its deacetylated derivative, N-(2'-deoxyguanosin-8-yl)-2-aminofluorene (dG-AF). Although the dG AAF and dG-AF adducts are distinguished only by the presence or absence of an acetyl group, they have profoundly different effects on DNA replication. dG-AAF poses a strong block to DNA synthesis and primarily induces frameshift mutations in bacteria, resulting in the loss of one or two nucleotides during replication past the lesion. dG-AF is less toxic and more easily bypassed by DNA polymerases, albeit with an increased frequency of misincorporation opposite the lesion, primarily resulting in G --> T transversions. We present three crystal structures of bacteriophage T7 DNA polymerase replication complexes, one with dG-AAF in the templating position and two others with dG-AF in the templating position. Our crystallographic data suggest why a dG-AAF adduct blocks replication more strongly than does a dG-AF adduct and provide a possible explanation for frameshift mutagenesis during replication bypass of a dG-AAF adduct. The dG-AAF nucleoside adopts a syn conformation that facilitates the intercalation of its fluorene ring into a hydrophobic pocket on the surface of the fingers subdomain and locks the fingers in an open, inactive conformation. In contrast, the dG-AF base at the templating position is not well defined by the electron density, consistent with weak binding to the polymerase and a possible interchange of this adduct between the syn and anti conformations. PMID- 15528278 TI - Regulatory actions of the A-kinase anchoring protein Yotiao on a heart potassium channel downstream of PKA phosphorylation. AB - A-kinase anchoring proteins (AKAPs) are thought to be passive members of protein complexes that coordinate the association of cAMP-dependent protein kinase A (PKA) with cellular substrates to facilitate targeted PKA protein phosphorylation. I(Ks), the slow heart potassium current, is carried by the I(Ks) potassium channel, a substrate for PKA phosphorylation in response to sympathetic nerve stimulation, is a macromolecular complex that includes the KCNQ1 alpha subunit, the KCNE1 regulatory subunit, and the AKAP Yotiao. Disruption of this regulation by mutation in the long QT syndrome is associated with elevated risk of sudden death. Here, we have studied the effects of the AKAP Yotiao on the function of the I(Ks) channel that had been mutated to simulate channel phosphorylation, and we report direct AKAP-mediated alteration of channel function distinct from its role in the coordination of channel phosphorylation by PKA. These data reveal previously undescribed actions of Yotiao that occur subsequent to channel phosphorylation and provide evidence that this adaptor protein also may serve as an effector in regulating this important ion channel. PMID- 15528279 TI - Hydrophobic interactions drive ligand-receptor recognition for activation and inhibition of staphylococcal quorum sensing. AB - Two-component systems represent the most widely used signaling paradigm in living organisms. Encoding the prototypical two-component system in Gram-positive bacteria, the staphylococcal agr (accessory gene regulator) operon uses a polytopic receptor, AgrC, activated by an autoinducing peptide (AIP), to coordinate quorum sensing with the global synthesis of virulence factors. The agr locus has undergone evolutionary divergence, resulting in the formation of several distinct inter- and intraspecies specificity groups, such that most cross group AIP-receptor interactions are mutually inhibitory. We have exploited this natural diversity by constructing and analyzing AgrC chimeras generated by exchange of intradomain segments between receptors of different agr groups. Functional chimeras fell into three general classes: receptors with broadened specificity, receptors with tightened specificity, and receptors that lack activation specificity. Testing of these chimeric receptors against a battery of AIP analogs localized the primary ligand recognition site to the receptor distal subdomain and revealed that the AIPs bind primarily to a putative hydrophobic pocket in the receptor. This binding is mediated by a highly conserved hydrophobic patch on the AIPs and is an absolute requirement for interactions in self-activation and cross-inhibition of the receptors. It is suggested that this recognition scheme provides the fundamental basis for agr activation and interference. PMID- 15528280 TI - Evaluation of the participation of Aretaieion Hospital, Greece in the WHO Pilot Project of Health Promoting Hospitals. AB - Aretaieion University Hospital participated in the World Health Organization (WHO) Pilot Health Promoting Hospitals Project from 1993 until 1997, when the Hellenic Network of Health Promoting Hospitals was established. This study is an overall evaluation of the participation of Aretaieion Hospital at the HPH Pilot Project. The evaluation comprised two questionnaire surveys, one among all the professionals, and the other among the patients involved in the subprojects. At the professional's survey, the response was 88.6% (39), while at the patient's survey the response was 62.2% (84). Differences were identified between the perceptions of the professionals who were external collaborators and those who were members of the hospital staff. Problems of the Project were reported to be lack of funding, lack of time and personnel shortages, lack of health promotion background and lack of incentives. Major successes were reported to be the personnel training, the development of collaboration and teamwork, the improvement of the overall image of the hospital into the community, the improvement of the working conditions of the personnel. Perception of sustainability of the projects by health professionals was found to be related to how interesting they found their subproject and how relevant to the work of their department. The communication strategy was inadequately planned and this resulted in some difficulties in developing rapport with the patients. However, the patients felt that they benefited from the project, a view also held by the hospital staff professionals. PMID- 15528281 TI - Early referral recommendations for ankylosing spondylitis (including pre radiographic and radiographic forms) in primary care. AB - An earlier diagnosis of ankylosing spondylitis (AS) is required because there is still a 5-7 year delay between first symptoms and diagnosis, and new effective treatments are available for active disease. Primary care physicians need easy to apply parameters to help them identify patients with suspected AS for onward referral. The best measures found were inflammatory back pain and HLA-B27 positivity. PMID- 15528282 TI - A naturalistic study of the determinants of health related quality of life improvement in osteoarthritic patients treated with non-specific non-steroidal anti-inflammatory drugs. AB - OBJECTIVES: To capture changes in the quality of life (QoL) occurring in patients with osteoarthritis (OA) during treatment with non-specific non-steroidal anti inflammatory drugs (NSAIDs) and to identify factors that predict such changes. METHODS: A naturalistic, prospective follow up of 783 patients with OA in whom primary care physicians decided to start treatment with non-selective NSAIDs. Short Form-36 (SF-36) and the Western Ontario and McMaster Universities OA index (WOMAC) were assessed at baseline and after 3 months. Baseline results were compared with QoL values in 4800 subjects randomly selected from the general population. Multiple regression analysis was performed to identify determinants of QoL at baseline and measures influencing changes in SF-36 or WOMAC during follow up. RESULTS: All QoL dimensions were significantly (p<0.01) decreased in patients with OA compared with controls. Significant improvement (p<0.05) in four dimensions of the SF-36 (vitality, role emotional, role physical, bodily pain) and in all components of the WOMAC was seen between baseline and month 3. Older age, female sex, longer duration of OA, and a higher number of comorbidities were the major determinants of a poor QoL at baseline. Maximal benefit from non specific NSAIDs was seen in patients with the most severe impairment in QoL and the shortest duration of OA. CONCLUSION: OA negatively impacts all dimensions of the QoL. Non-specific NSAIDs improve the QoL in patients with OA treated in a "real life setting". The profile of patients receiving maximal benefit from such treatment may be of interest for health providers, enabling them to decide who should preferentially be given cytoprotective treatments or coxibs. PMID- 15528283 TI - A randomised, double blind, placebo controlled, multicentre trial of combination therapy with methotrexate plus ciclosporin in patients with active psoriatic arthritis. AB - OBJECTIVES: To evaluate the safety and efficacy of adding ciclosporin A (CSA) to the treatment of patients with psoriatic arthritis (PsA) demonstrating an incomplete response to methotrexate (MTX) monotherapy. METHODS: In a 12 month, randomised, double blind, placebo controlled trial at five centres in three countries, 72 patients with active PsA with an incomplete response to MTX were randomised to receive either CSA (n = 38) or placebo (n = 34). Patients underwent full clinical and radiological assessment and, in addition, high resolution ultrasound (HRUS) was performed at one centre. An intention to treat (last observation carried forward) analysis was employed. RESULTS: Some significant improvements were noted at 12 months in both groups. However, in the active but not the placebo arm there were significant improvements in swollen joint count, mean (SD), from 11.7 (9.7) to 6.7 (6.5) (p<0.001) and C reactive protein, from 17.4 (14.5) to 12.7 (14.3) mg/l (p<0.05) as compared with baseline. The Psoriasis Area and Severity Index (PASI) score improved in the active group (2 (2.3) to 0.8 (1.3)) as compared with placebo (2.2 (2.7) to 1.9 (2.8)), p<0.001, and synovitis detected by HRUS (33 patients, 285 joints) was reduced by 33% in the active group compared with 6% in the placebo group (p<0.05). No improvement in Health Assessment Questionnaire or pain scores was detected. CONCLUSIONS: Synovitis detected by HRUS was significantly reduced. Combining CSA and MTX treatment in patients with active PsA, and a partial response to MTX, significantly improves the signs of inflammation but not pain or quality of life. PMID- 15528284 TI - Preferences of patients for emergency services available during usual GP surgery hours: a discrete choice experiment. AB - OBJECTIVE: A study was undertaken to investigate patients' strength of preferences for attributes or characteristics associated with delivery of emergency primary care services available during usual GP surgery hours and to investigate the trade-offs between attributes. METHODS: A discrete choice experiment was used to quantify patients' strength of preferences for several key attributes of usual-hours emergency primary care. The attributes were chosen to reflect the findings of previous research, current policy initiatives and discussions with local key stakeholders. A self-complete questionnaire was administered to NHS Direct callers and adult attenders at Accident and Emergency, GP services and the NHS Walk-in Centre in the locality. Regression analysis was used to estimate the relative importance to patients of the different attributes. RESULTS: An overall response of 71% (n=432) was achieved. All but one of the attributes was a statistically significant predictor of preference. The attribute 'being kept informed about waiting time' was the most important. This was followed by 'quality of the consultation', 'having a consultation with a nurse', 'having a consultation with a doctor' and 'contacting the service in person'. Respondents were prepared to wait an extra 68 min to have a consultation with a doctor, but an extra 2 h 9 min for information about expected waiting time. There were no measurable preference differences between patients surveyed at different NHS entry points. Respondents younger than 45 years held strong preferences with respect to how they wanted to make contact with the system, whereas older respondents appeared not to hold strong preferences, seemingly indifferent between the alternatives. There was weak evidence which showed the younger group more strongly preferred accessing services via an integrated telephone system than making contact in person. CONCLUSIONS: This study showed that local solutions for reforming emergency primary care during hours when the GP surgery is open should take account of the strength of patient preferences. The discrete choice method was acceptable, and the results directly informed the development of a local service framework for such care. PMID- 15528285 TI - What prevents older people from seeking treatment for urinary incontinence? A qualitative exploration of barriers to the use of community continence services. AB - BACKGROUND: Urinary incontinence is a significant health problem for older people. Many people with incontinence do not seek services. Simple and effective treatments exist in primary care. OBJECTIVES: Our aim was to explore reasons why older people living in the community do not present for help with problems of urinary incontinence and to identify ways in which they may be assisted to access continence services. METHODS: In-depth interviews lasting an average of 1 h were conducted with 20 people aged over 65 years living in the community, purposively selected from a sample of patients who volunteered to be interviewed. RESULTS: Older people described ageing as a natural, degenerative process and had reduced health expectations. Urinary incontinence was commonly viewed as an inevitable aspect of ageing and, as such, something to be accepted and managed independently. Shame and embarrassment combined with generational differences in attitudes to disclosure about personal matters also prevented people from seeking advice. Relatively successful strategies to manage incontinence enabled people to contain their symptoms, although this was at a social, psychological and, in some cases, health cost. Older people in this sample had considerable co-morbidity, in many cases resulting in mobility problems. Despite regular contact with primary care professionals, they had seldom disclosed their urinary incontinence. CONCLUSION: A combination of personal attitudes and practical barriers prevent older people from seeking help for urinary incontinence. It is possible that older people would be more likely to seek help if asked specifically about urinary leakage by primary health care professionals. PMID- 15528286 TI - The consultation and relational empathy (CARE) measure: development and preliminary validation and reliability of an empathy-based consultation process measure. AB - BACKGROUND: Empathy is a key aspect of the clinical encounter but there is a lack of patient-assessed measures suitable for general clinical settings. OBJECTIVES: Our aim was to develop a consultation process measure based on a broad definition of empathy, which is meaningful to patients irrespective of their socio-economic background. METHODS: Qualitative and quantitative approaches were used to develop and validate the new measure, which we have called the consultation and relational empathy (CARE) measure. Concurrent validity was assessed by correlational analysis against other validated measures in a series of three pilot studies in general practice (in areas of high or low socio-economic deprivation). Face and content validity was investigated by 43 interviews with patients from both types of areas, and by feedback from GPs and expert researchers in the field. RESULTS: The initial version of the new measure (pilot 1; high deprivation practice) correlated strongly (r = 0.85) with the Reynolds empathy measure (RES) and the Barrett-Lennard empathy subscale (BLESS) (r = 0.63), but had a highly skewed distribution (skew -1.879, kurtosis 3.563). Statistical analysis, and feedback from the 20 patients interviewed, the GPs and the expert researchers, led to a number of modifications. The revised, second version of the CARE measure, tested in an area of low deprivation (pilot 2) also correlated strongly with the established empathy measures (r = 0.84 versus RES and r = 0.77 versus BLESS) but had a less skewed distribution (skew -0.634, kurtosis -0.067). Internal reliability of the revised version was high (Cronbach's alpha 0.92). Patient feedback at interview (n = 13) led to only minor modification. The final version of the CARE measure, tested in pilot 3 (high deprivation practice) confirmed the validation with the other empathy measures (r = 0.85 versus RES and r = 0.84 versus BLESS) and the face validity (feedback from 10 patients). CONCLUSIONS: These preliminary results support the validity and reliability of the CARE measure as a tool for measuring patients' perceptions of relational empathy in the consultation. PMID- 15528287 TI - The association between maternal ratings of child health interviews and maternal and child characteristics. AB - BACKGROUND: Doctors develop the skills needed to interview parents and children in paediatric settings by practice and by receiving feedback during their medical training. Interviewed parents are ideally placed to provide evaluations of these skills. If parents, as consumers of health care services, are to be consulted, it is important to determine whether factors other than interview skills affect their evaluations. OBJECTIVES: Our aim was to examine the relationship between maternal satisfaction ratings of student doctor interviews, and maternal and child characteristics. METHODS: Sixty mothers of children attending the paediatric medical out-patient clinic at the Women's and Children's Hospital, South Australia were allocated randomly to rate one of four video-taped final year student doctor interviews (15 mothers per interview). The level of skills displayed by the student doctor differed in each interview. Maternal satisfaction was measured using the Medical Interview Satisfaction Scale (MISS) and the Interpersonal Skills Rating Scale (IPS), and interview ratings were compared for a number of maternal and child characteristics. RESULTS: No significant associations were observed between maternal satisfaction ratings and any maternal or child characteristics other than lower satisfaction associated with previous experience of a real student doctor interview (P <0.01). The interview seen by mothers predicted 53% (MISS) and 65% (IPS) of the variance in maternal satisfaction ratings. After controlling for the interview type, the maternal and child characteristics studied predicted 17% additional variance in MISS scores and 7% in IPS scores. CONCLUSION: The quality of the interview skills demonstrated was the principle determinant of maternal satisfaction ratings. PMID- 15528288 TI - The effect of health status on patients' satisfaction with out-of-hours care provided by a family doctor co-operative. AB - BACKGROUND: Systems for providing primary care outside normal hours have changed significantly in Europe over the last 20 years. The impetus for this change has come almost entirely from the medical profession, and it is important to consider the patients' perspective. Although patient's satisfaction with out-of-hours care has been studied extensively, the effect of patient's health status on satisfaction level has not been examined previously. OBJECTIVES: The primary objective of this study was to investigate whether health status has an influence on patient satisfaction with out-of-hours care provided by a family doctor co operative. The secondary objective of this study was to investigate the impact of age, gender, socio-economic status and call outcome on patients' satisfaction with out-of-hours care. METHODS: All patients contacting the service over a designated 24 day period were forwarded a postal questionnaire. Health status was recorded using the Short Form-12 (SF-12) health survey. Patients' satisfaction was measured by using a version of the McKinley questionnaire. RESULTS: The response rate was 55% (531 out of 966). Overall satisfaction levels were high, with 88% of patients rating the service as either excellent or good. Logistic regression, modelling for the simultaneous effects of age, gender, socio-economic status, call outcome and health status on overall satisfaction, found that patients with lower physical and mental health status scores were significantly less likely to be satisfied with their out-of-hours care [odds ratio (OR) 1.04, 95% confidence interval (CI) 1.01-1.07, P = 0.017; and 1.03, 95% CI 1.00-1.06, P = 0.046, respectively]. Patients with higher socio-economic status were also significantly less likely to be satisfied (OR 0.25, 95% CI 0.11-0.55, P = 0.001). Patient's age and gender, and call outcome did not significantly affect overall satisfaction levels. CONCLUSION: Family doctor co-operatives have significantly altered the way out-of-hours care is delivered. Patients with lower health status are significantly less likely to be satisfied with this new form of out-of-hours care. This finding has important implications for the future planning of out-of hours primary care services. PMID- 15528289 TI - Factors affecting feasibility and acceptability of a practice-based educational intervention to support evidence-based prescribing: a qualitative study. AB - BACKGROUND: Inappropriate and costly GP prescribing is a major problem facing Primary Care Trusts. Educational outreach into practices, alongside other measures, such as audit and feedback, have the potential to enable GP prescribing to become more evidence based. High GP prescribing costs are associated with GPs who see drug company representatives; tend to end consultations with prescriptions; and 'try out' new drugs on an 'ad hoc basis' and use this as evidence of the drug's effect. An educational intervention called 'reflective practice' was developed to meet these and other educational needs. The design of the intervention was informed by studies that have identified the pre-requisites of successful behaviour change in general practice. OBJECTIVES: The study investigated the following: (i) Is it feasible for GPs to attend the sessions included in the educational intervention? (ii) Is the intervention acceptable to the participants and the session facilitators? (iii) What are the barriers to the group educational processes, and how can these be overcome? METHODS: Four practices were recruited in South West England, all of them experiencing problems with prescribing appropriateness and cost. Reflective practice sessions (including a video-taped scenario) were run in each of these practices and qualitative methods were used to explore the complex attitudes and behaviour of the participants. A researcher observed and audio-taped sessions in each practice. At the end of the programme, a sample of doctors and all the facilitators were interviewed about their experiences. The recorded data were transcribed and analysed using standard qualitative methods. RESULTS: The doctors in the largest partnerships were those who had the greatest difficulty in attending the sessions. Elsewhere, doctors were also reluctant to become involved because of previous experience of top-down managerial initiatives about prescribing quality. Facilitators came from a broad range of professional backgrounds. While knowledge of prescribed drug management issues was important, the professional background of the facilitator was less important than group facilitation skills in creating a group process which participating GPs found satisfactory. The video-taped scenario was found to be useful to set the scene for the discussion. Preserving the anonymity of responses of the GPs in the initial stages of the sessions was important in ensuring honesty in the discussion. Reaching a consensus on management of common conditions was sometimes difficult, partly because the use of the term 'best buy' implies economic pressures, rather than benefits to patients, and partly because of the value with which GPs regard the concept of clinical autonomy. 'Reflective Practice' appeared to have the potential to make GPs aware of the link to be made between their clinical management decisions and the evidence provided by the British National Formulary and Clinical Evidence. CONCLUSION: The study indicates the importance of preparing the practice adequately, including providing protected time for all GPs to attend the educational intervention. Scenarios and the structure of the sessions need to make more explicit the links between everyday practice and published evidence of effectiveness. Emphasis on cost-effectiveness may be counterproductive and wider benefits need to be emphasized. We have also identified the skill profile of the facilitator role. Our study indicates a need for a clearer understanding of GPs' perception of clinical autonomy and how this conflicts with the goal of agreement on practice guidelines for treatment. The intervention is now ripe for further development, perhaps by integrating it with other interventions to change professional behaviour. The improved intervention should then be evaluated in a randomized controlled trial. PMID- 15528290 TI - South Asian patients' views and experiences of clinical trial participation. AB - OBJECTIVES: This paper explores South Asian patients' views and experiences of clinical trial participation, as part of a larger study which sought to assess British South Asian under-representation in clinical trials. METHODS: The study was based on semi-structured interviews with South Asian trial participants in primary care and specialist treatment centres in the north of England. Fifteen South Asian patients who had participated in one of six different clinical trials to test pharmaceutical products comprised the study cohort. RESULTS: South Asian motivations to participate in clinical trials are similar to those of the majority 'White' population. Clear and concise trial information (provided by caring and understanding trial staff) was considered an important aspect of the respondents' experiences. Appealing to South Asian peoples' altruistic nature by making them aware of South Asian under-representation (especially in clinical trials that investigate illnesses prevalent in their community) was also identified as a strong motivational factor. Potential barriers to their participation included: trial burden (which bears heavily on the poor), language, and discriminatory practices in the NHS, which can lead to mistrust of health professionals. In addition, female modesty and preference for female trial staff was recognized as a 'cultural' barrier to participation. CONCLUSION: There are more similarities than differences between the experiences of British South Asians and 'White' trial participants. Present findings suggest that ethnicity operates at different levels. In addition to South Asian trial participants' culture, social class and gender are also as likely to affect their participation in clinical trials. To improve South Asian accrual rates, strategies should be designed to take into consideration linguistic differences and improving overall trust in the clinical trial team. PMID- 15528291 TI - Patient-clinician agreement on signs and symptoms of 'strep throat': a MetroNet study. AB - BACKGROUND: Despite substantial use of the telephone in health care, only a few studies have formally evaluated the appropriateness of telephone-based management for acute medical problems. The accuracy of patients' report of signs and symptoms remains unknown. OBJECTIVE: We compared the agreement between patient self-assessment and clinician assessment on the typical signs and symptoms of group A beta-haemolytic Streptococcus (GABHS) to investigate the potential difficulties of using patient self-report to triage sore throat patients. METHODS: In this cross-sectional study, each of 200 adult pharyngitis patients was instructed to examine him/herself and to record the symptoms and physical findings. Two clinicians independently interviewed and examined each patient and recorded their findings. Each patient then had a rapid GABHS antigen test, the results of which were blinded to both clinicians and patients. Each patient self assessment was compared with the findings of each clinician, and the agreement and disagreement between them computed. RESULTS: We found varying levels of agreement (kappa=-0.05 to 0.71) between patients and clinicians on sore throat history and physical assessments. Importantly, there was fair to substantial agreement (kappa=0.20-0.71) on the key signs and symptoms used in GABHS clinical prediction rules. As expected, history items had the highest agreement (kappa=0.52-0.71). Patients were more likely than clinicians to report rather than deny a specific physical sign. CONCLUSION: Adult sore throat patients may reliably report their symptoms, but may not be able to assess and report accurately on relevant physical signs of pharyngitis. Patients have a tendency to over-report physical signs. This study indicates the potential difficulties associated with telephone triage of sore throat patients, or other illnesses that require assessment of physical signs. PMID- 15528292 TI - Electrocardiogram interpretation in general practice. AB - BACKGROUND: The 12-lead electrocardiogram (ECG) is a common diagnostic test available to the GP in the evaluation of patients with cardiac complaints. In daily clinical practice it is important for GPs to know the sensitivity and specificity of their ECG interpretation skills. OBJECTIVES: The purpose of the present study was to evaluate the ECG interpretation skills of GPs and the value of automatic ECG recorder interpretations in general practice. METHODS: A total of 902 ECGs were recorded in a random sample of the population aged 31-51 years in the district of Ebeltoft, Denmark, from December 1991 to June 1992. They were interpreted automatically by an interpretive ECG recorder and by the GPs in the clinic in Ebeltoft, with a cardiologists interpretation as a gold standard. Sensitivity, specificity and predictive values of diagnoses were calculated. RESULTS: Overall, the sensitivity of abnormal diagnoses made by the GPs (69.8%) was significantly lower (P <0.001) than that of diagnoses made by the interpretive ECG recorder (84.4%). The overall specificity of abnormal diagnoses made by the GP (85.7%) was significantly higher (P <0.001) than that achieved by the interpretive ECG recorder (75.6%). CONCLUSIONS: GPs in this study were good at correcting false-positive diagnoses made by the interpretive ECG recorder. In order to avoid unfortunate reclassifications of true-positive to false-negative diagnoses, GPs are recommended to pay special attention to the diagnoses of ST segment deviation, T-wave inversion or the presence of Q-waves made by interpretive ECG recorders, when ECGs are used in individual risk assessment. PMID- 15528293 TI - Changes of cell-free fetal DNA in maternal plasma after elective termination of pregnancy. PMID- 15528294 TI - Response of homocysteine, cystathionine, and methylmalonic acid to vitamin treatment in dialysis patients. AB - BACKGROUND: Hyperhomocysteinemia is observed in >80% of hemodialysis patients and is considered a risk factor for cardiovascular disease. Vitamin treatment lowers total homocysteine (tHcy) concentrations in plasma and may therefore reduce the associated risk. Current treatment strategies have not achieved normalization of tHcy in the majority of dialysis patients. METHODS: We administered folic acid (5 mg) plus vitamin B(6) (50 mg) and B(12) (0.7 mg) intravenously to 38 hyperhomocysteinemic patients (tHcy >18 micromol/L) after each dialysis treatment. The treatment phase lasted 1 month, and serum concentrations of tHcy, methylmalonic acid (MMA), and cystathionine were measured at weeks 0, 2, 4, 6, 8, and 24. RESULTS: The median serum tHcy concentration decreased significantly, from 26.1 micromol/L at baseline to 13.2 micromol/L at week 4. The median change in tHcy after 4 weeks was 13.4 micromol/L (-51%) compared with baseline. Serum MMA and cystathionine concentrations were reduced by 28% and 26%, respectively, but neither was normalized at 4 weeks. Backward-elimination stepwise regression analysis revealed that higher concentrations of tHcy, MMA, and cystathionine and lower folate at baseline predict changes of tHcy after treatment. Twenty weeks after vitamin withdrawal, tHcy concentrations returned to values comparable to baseline (median, 24.8 micromol/L). CONCLUSIONS: The combination of folic acid, vitamin B(12), and vitamin B(6) used in this study normalized serum concentrations of tHcy in almost all of our hyperhomocysteinemic dialysis patients. This regimen may be used to investigate the effects of homocysteine normalization on cardiovascular outcomes in hemodialysis patients. PMID- 15528295 TI - The novel apolipoprotein A5 is present in human serum, is associated with VLDL, HDL, and chylomicrons, and circulates at very low concentrations compared with other apolipoproteins. AB - BACKGROUND: The recently discovered apolipoprotein A5 (ApoA5) is fast gaining attention as a key regulator of serum triglyceride concentrations. An ApoA5 mouse knock-out model produced an approximately fourfold increase in serum triglycerides, whereas a knock-in model with human ApoA5 produced 50-70% lower concentrations of mouse serum triglycerides. In addition, peroxisome proliferator activated receptor-alpha agonists, which are used clinically to lower serum triglyceride concentrations, cause increased ApoA5 mRNA expression. Despite these compelling molecular biology data, relatively little is known about ApoA5 protein in human serum. METHODS: To better understand circulating concentrations and lipoprotein particle distribution of ApoA5, we expressed the recombinant human ApoA5 protein and raised antibodies against both the NH(2) and COOH termini. RESULTS: Using the above reagents, we demonstrate for the first time that ApoA5 is present in human serum, although at much lower concentrations than other apolipoproteins such as ApoA1. Using a dual-antibody sandwich ELISA that we developed, we observed ApoA5 concentrations in human serum ranging from 24 to 406 microg/L compared with approximately 1 g/L for ApoA1. We also examined the lipoprotein particle distribution of ApoA5 and found that ApoA5 was detectable in VLDL, HDL, and chylomicrons, but not LDL. CONCLUSIONS: These data demonstrate for the first time that ApoA5 is a secreted protein present in human serum and is associated with specific lipoprotein particles. In addition, our data indicate that the circulating concentration of human ApoA5 is very low compared with other apolipoproteins. PMID- 15528296 TI - Characterization of novel monoclonal antibodies for prostate-specific antigen (PSA) with potency to recognize PSA bound to alpha 2-macroglobulin. AB - BACKGROUND: Different molecular forms of prostate-specific antigen (PSA) have been used to differentiate between benign prostatic hyperplasia and prostate cancer. Detecting PSA bound to endogenous inhibitors such as alpha(1) antichymotrypsin (ACT) and alpha(2)-macroglobulin (alpha(2)M) is often difficult because of epitope masking or sensitivity problems. Here we report the characterization of four novel mouse monoclonal antibodies (mabs) obtained by immunization with PSA-alpha(2)M complexes. Their ability to detect free PSA and PSA-inhibitor complexes was shown, and their epitopes were analyzed by phage display technology. METHODS: The properties of the mabs were studied by competition and sandwich assays and by Western blotting. Epitope mapping was performed by screening of a phage display peptide library. RESULTS: All four mabs recognized free PSA, PSA-ACT, and PSA-alpha(2)M complexes, but to various degrees. With different combinations of mabs in competition experiments, antibodies were identified that enhance binding of other mabs to PSA, forming the molecular basis of a very sensitive assay for the detection of PSA and PSA-ACT complexes. Mabs with highest reactivity for PSA-alpha(2)M were selected to establish an immunoassay for that complex. Western blot analysis revealed that all mabs recognized conformational epitopes of PSA. These findings were supported by phage display results demonstrating mimotopes in the PSA molecule. CONCLUSION: The results presented here could aid in the further development of clinically relevant assays for PSA and PSA-alpha(2)M complexes. PMID- 15528297 TI - Perturbation of NgTRF1 expression induces apoptosis-like cell death in tobacco BY 2 cells and implicates NgTRF1 in the control of telomere length and stability. AB - Telomeres are specialized nucleoprotein complexes that are essential for preserving chromosome integrity in eukaryotic cells. Several potential telomere binding proteins have recently been identified in higher plants, but nothing is known about their in vivo functions. We previously identified NgTRF1 as a double stranded telomeric repeat binding factor in tobacco (Nicotiana tabacum) and here show that the binding of NgTRF1 to telomeric repeats inhibits telomerase-mediated telomere extension. To determine whether NgTRF1 is involved in telomere length regulation, we established transgenic tobacco BY-2 cell lines that overexpress or suppress NgTRF1. Pulsed-field gel electrophoresis showed that 35S::NgTRF1 cells exhibited significantly shortened telomeres (45 to 10 kb), whereas 35S::antisense NgTRF1 cells contained longer telomeres (80 to 25 kb) compared with wild-type and 35S::GUS control cells (65 to 15 kb), indicating that telomere length inversely correlates with the amount of functional NgTRF1 in BY-2 cells. 35S::NgTRF1 cells with shorter telomeres displayed a progressive reduction in cell viability and stopped dividing after 25 to 40 successive rounds of 12-d batch subculture, in sharp contrast with control cells, which have an unlimited capacity for division. Internucleosomal DNA fragmentation, mitochondrial release of cytochrome c, and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling positive nuclei were detected in 35S::NgTRF1 cells during prolonged subculture, indicating that enhanced cell death was attributable to an apoptosis-like mechanism. 35S::antisense-NgTRF1 cells containing low levels of NgTRF1 also exhibited a progressive decrease in cell viability and apoptotic cell death, but less so than did 35S::NgTRF1 cells, suggesting that the level of NgTRF1 is critically associated with cell viability. Taken together, these data indicate that perturbation of NgTRF1 expression results in changes in telomere length and stability, which in turn causes apoptotic cell death in transgenic BY-2 cells. These results are discussed in light of the suggestion that NgTRF1 is involved in the mechanism by which telomere length and stability are maintained. We further suggest that the structural stability of telomeres, in addition to length maintenance, is essential for their function and for the immortality of BY-2 cells. PMID- 15528298 TI - Structural insights into the target specificity of plant invertase and pectin methylesterase inhibitory proteins. AB - Pectin methylesterase (PME) and invertase are key enzymes in plant carbohydrate metabolism. Inhibitors of both enzymes constitute a sequence family of extracellular proteins. Members of this family are selectively targeted toward either PME or invertase. In a comparative structural approach we have studied how this target specificity is implemented on homologous sequences. By extending crystallographic work on the invertase inhibitor Nt-CIF to a pectin methylesterase inhibitor (PMEI) from Arabidopsis thaliana, we show an alpha helical hairpin motif to be an independent and mobile structural entity in PMEI. Removal of this hairpin fully inactivates the inhibitor. A chimera composed of the alpha-hairpin of PMEI and the four-helix bundle of Nt-CIF is still active against PME. By contrast, combining the corresponding segment of Nt-CIF with the four-helix bundle of PMEI renders the protein inactive toward either PME or invertase. Our experiments provide insight in how these homologous inhibitors can make differential use of similar structural modules to achieve distinct functions. Integrating our results with previous findings, we present a model for the PME-PMEI complex with important implications. PMID- 15528299 TI - Cross-Talk between Fas/Fas ligand system and nitric oxide in the pathway subserving granulosa cell apoptosis: a possible regulatory mechanism for ovarian follicle atresia. AB - Recent studies have shown the involvement of Fas/Fas ligand (FasL) system and nitric oxide (NO) in ovarian follicle atresia. Here we asked whether Fas/Fas ligand system interacts with NO using rat granulosa cell culture. Soluble recombinant Fas ligand (rFasL), at 100 ng/ml, significantly decreased cell viability, as measured by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl) 2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay, in the presence of 200 U/ml interferon-gamma, whereas the concurrent addition of a caspase inhibitor, Z VAD-FMK, at 20 microm, significantly inhibited rFasL-induced cytotoxicity. Hoechst 33342 staining and flow cytometric analysis confirmed the induction of apoptosis in granulosa cells by 100 ng/ml rFasL in the presence of interferon gamma, which was blocked by the concomitant addition of an NO donor, S-nitroso-N acetylpenicillamine. Western blot analysis demonstrated that rFasL significantly up-regulated caspase-3, -8, and -9 activities in granulosa cells, which were attenuated by concurrent treatment with S-nitroso-N-acetylpenicillamine. Real time quantitative RT-PCR revealed a significant decrease in inducible NO synthase mRNA levels in rFasL-induced apoptotic granulosa cells. In conclusion, we demonstrated the involvement of Fas/FasL system in inducing apoptosis through activation of a caspase-mediated cascade in rat granulosa cells, which is coupled with a decrease in inducible NO synthase expression. We further showed that NO inhibited Fas/FasL system-induced apoptosis by suppressing activation of the caspases, pointing to a cross-talk between Fas/FasL system-induced apoptosis pathway and NO-mediated antiapoptotic pathway in ovarian follicle atresia. PMID- 15528300 TI - Progesterone antagonizes the permissive action of estradiol on tumor necrosis factor-alpha-induced apoptosis of anterior pituitary cells. AB - We previously reported that TNF-alpha-induced apoptosis of lactotropes is estrogen dependent and predominant at proestrus. Here we observed that TNF-alpha (50 ng/ml) failed to induce apoptosis of anterior pituitary cells from ovariectomized rats cultured in the presence of progesterone (10(-6) m). However, progesterone blocked the apoptotic effect of TNF-alpha in anterior pituitary cells and lactotropes cultured with 17beta-estradiol (10(-9) m). In addition, 17beta-estradiol induced apoptosis of somatotropes and triggered the proapoptotic action of TNF-alpha in these cells, effects completely blocked by ICI 182 780 (10(-6) m), an estrogen receptor antagonist. Progesterone reverted the permissive effect of 17beta-estradiol on TNF-alpha-induced apoptosis of somatotropes. TNF alpha induced apoptosis of somatotropes from rats killed at proestrus but not at diestrus. The antiprogestine ZK 98,299 (10(-6) m) completely inhibited the protective action of progesterone on TNF-alpha-induced apoptosis of anterior pituitary cells, lactotropes, and somatotropes. Although progesterone can interact with glucocorticoid receptors, dexamethasone (10(-6) m) had no effect on TNF-alpha-induced apoptosis of anterior pituitary cells, lactotropes, and somatotropes. Our results show that progesterone, by interacting with progesterone receptors, antagonizes the permissive action of estrogens on TNF alpha-induced apoptosis of lactotropes and somatotropes. These observations suggest that the steroid milieu may modulate the apoptotic response of anterior pituitary cells during the estrous cycle. PMID- 15528301 TI - Transcriptional and posttranscriptional regulation of fibulin-1 by estrogens leads to differential induction of messenger ribonucleic acid variants in ovarian and breast cancer cells. AB - Fibulin-1 is an extracellular matrix protein overexpressed in epithelial ovarian and breast cancers. In estrogen receptor (ER)-positive ovarian and breast cancer cell lines, fibulin-1 mRNA levels are markedly increased by estrogens. Transfection experiments using fibulin-1 promoter constructs indicate that 17beta estradiol (E2) increases fibulin-1 gene transcription and that ERalpha is more potent than ERbeta to mediate E2 regulation of the transfected fibulin-1 promoter. Using SL2 cells devoid of specificity protein 1 (Sp1) and site-directed mutagenesis of GC boxes, we evidenced that the E2 regulation occurs through a proximal specificity protein 1 binding site. In addition, we show that fibulin-1C and -1D mRNAs, the two major fibulin-1 splicing variants, are differentially induced by E2. The induction of both mRNAs variants is direct and independent of a newly synthesized protein intermediate. Interestingly, actinomycin D chase experiments demonstrate that E2 treatment selectively shortens the fibulin-1D mRNA half-life. This indicates that estrogens affect differentially the stability of fibulin-1 variants and may explain the lower accumulation of fibulin-1D mRNA on E2 treatment. In conclusion, our data show that estrogens, via ERalpha, are key regulators of fibulin-1 expression at both the transcriptional and posttranscriptional levels. The preferential induction of the fibulin-1C variant, which is overexpressed in ovarian and breast cancer, might play an important role in estrogen-promoted carcinogenesis. PMID- 15528302 TI - Interferon stimulated gene 15 conjugates to endometrial cytosolic proteins and is expressed at the uterine-placental interface throughout pregnancy in sheep. AB - Interferon-stimulated gene 15 (ISG15) is a ubiquitin homolog expressed in uteri of ruminants in response to interferon (IFN)-tau and is also induced during pregnancy in the uteri of mice, pigs, humans, and baboons. This study examined expression of ISG15 and its conjugation to target proteins in the ovine uterus beyond the period of IFNtau secretion by the conceptus. Although steady-state levels of ISG15 mRNA decreased after d 25 of pregnancy, ISG15 persisted in endometrium through d 120. In situ hybridization and immunocytochemistry localized ISG15 across the entire uterine wall through d 25, after which expression was restricted to endometrial stroma along the maternal-placental interface. Western blots revealed ISG15 and ISG15-conjugated proteins in endometrium. Treatment of ovariectomized sheep with progesterone and IFNtau increased both free and conjugated ISG15. These results are the first to show in vivo regulation of ISG15 function (i.e. conjugation to target proteins) by a type I IFN in the uterus of any species and that ISG15 is expressed at contacts between the placenta and uterus when trophectoderm no longer produces IFNtau. Interestingly, mRNA for the type II IFNgamma was present in the endometrial stromal compartment on d 15-50, which may stimulate the synthesis of ISG15 through later pregnancy. We hypothesize that ISG15 is not merely a consequence of an antiviral state induced by trophoblast IFNtau but represents a critical component of the microenvironment at the uterine-placental interface during the progressive events of conceptus development, implantation, and placentation in sheep and perhaps other mammalian species. PMID- 15528303 TI - Significance of prohormone convertase 2, PC2, mediated initial cleavage at the proglucagon interdomain site, Lys70-Arg71, to generate glucagon. AB - To define the biological significance of the initial cleavage at the proglucagon (PG) interdomain site, K70-R71 downward arrow, we created two interdomain mutants, K70Q-R71Q and R71A. Cotransfection studies in GH4C1 cells show significant amounts of glucagon production by PC2 along with some glicentin, glicentin-related polypeptide-glucagon (GRPP-glucagon) and oxyntomodulin from wild-type PG. In contrast, a larger peptide, PG 33-158, and low amounts of GRPP glucagon are predominantly generated from interdomain mutants. HPLC analysis shows a 5-fold increase in glucagon production by PC2 from wild-type PG and a corresponding 4-fold lower accumulation and secretion of unprocessed precursor relative to interdomain mutants. PC2 generates significant levels of glucagon from a glicentin (PG 1-69) expression plasmid, whereas PC1/3 produces only modest amounts of oxyntomodulin. Employing a major PG fragment (PG 72-158) expression plasmid, we show that PC1/3 predominantly generates glucagon-like peptide (GLP) 1, whereas PC2 produces only N-terminally extended GLP-1. Surprisingly, production of GLP-1 and GLP-2 by PC1/3 from interdomain mutants, compared with wild-type PG, is not significantly impaired. In addition to PC2 and PC1/3, PC5/6A and furin are also able to cleave the sites, K70-R71 downward arrow and R107-X-R R110 downward arrow in PG. We show a much greater ability of furin to cleave the monobasic site, R77 downward arrow, than at the dibasic site, R124-R125 downward arrow, which is also weakly processed by PC5/6A, indicating overlapping specificities of these two convertases mainly with PC1/3. We propose here a trimer-like model of the spatial organization of the hormonal sequences within the PG molecule in which the accessibility to prohormone convertase action of most cleavage sites is restricted with the exception of the interdomain site, K70 R71, which is maximally accessible. PMID- 15528304 TI - Generation of globular fragment of adiponectin by leukocyte elastase secreted by monocytic cell line THP-1. AB - Previous studies revealed that carboxyl-terminal fragment containing the globular domain of adiponectin exists in human plasma. Although it is proposed that the globular fragment is generated by proteolytic cleavage, the place and responsible enzyme of the cleavage are still unclear. In this study, we evaluated the activity to cleave adiponectin in culture medium of several cell lines in vitro. Adiponectin cleavage into several carboxyl-terminal fragments containing the globular domain was observed in the medium of phorbol 12-myristate 13-acetate stimulated monocytic cell lines THP-1 and U937. The molecular masses of the major products were 25, 20, and 18 kDa. The cleavage was thought to be mediated by leukocyte elastase (also known as neutrophil elastase) based on the following observations. First, the cleavage was inhibited by serine-protease inhibitors [phenylmethylsulfonylfluoride, Pefabloc SC (Roche Diagnostics, Basel, Switzerland) and aprotinin] and by the leukocyte elastase-specific peptide inhibitor MeOSuc-AAPV-CMK. Second, no activity was detected after THP-1 cells had fully differentiated into macrophages. Third, purified leukocyte elastase cleaved adiponectin with the same cleavage pattern as THP-1 cells. Finally, leukocyte elastase secreted by activated neutrophils cleaved adiponectin into the globular fragments. Amino-terminal sequence analysis revealed that cleavage sites of adiponectin by purified leukocyte elastase were between 38Thr and 39Cys, 40Ala and 41Gly, 44Ala and 45Gly, 91Ala and 92Glu, and 110Ala and 111Ala (the numbering of the positions of the amino acids starts at the signal sequence), suggesting that the cleavage occurs in the collagenous domain. These data indicate that the cleavage of adiponectin by leukocyte elastase secreted from activated monocytes and/or neutrophils could be a candidate for the mechanism of the generation of the globular fragment of adiponectin. PMID- 15528305 TI - Y2 receptor-selective agonist delays the estrogen-induced luteinizing hormone surge in ovariectomized ewes, but y1-receptor-selective agonist stimulates voluntary food intake. AB - Neuropeptide Y (NPY) plays a major role in the regulation of food intake, regulation of homeostasis, and neuroendocrine function. We have previously shown that third ventricular infusion of this peptide delays the estradiol benzoate induced surge in LH secretion in ovariectomized ewes. To determine the receptor subtype that transmits this effect, we have now used the same model to infuse a Y1 receptor agonist [NPY Leu31 Pro34], a Y2 receptor agonist (PYY3-36), and a Y4 receptor agonist (pancreatic polypeptide). We monitored the surges in animals given these agonists or artificial cerebrospinal fluid by measuring plasma LH levels, and we also measured daily voluntary food intake (VFI). A low (7 microg/h) dose of Y2 agonist delayed the surge but did not affect VFI, whereas a higher dose (14 microg/h) stimulated VFI. A dose of 18 microg/h of the Y1 agonist did not affect surge generation but also stimulated VFI. A dose of 24 microg/h of Y4 agonist affected neither surge generation nor VFI. These specificities are different from those reported for the rat and human (in which a Y2 agonist causes reduction in VFI). We conclude that, in sheep, the negative regulation of the reproductive axis by NPY and Y-receptor agonists is effected via the Y2 receptors, whereas the orexigenic effects are most likely effected via the Y1 receptors. PMID- 15528306 TI - Hydrogen peroxide is essential for estrogen-deficiency bone loss and osteoclast formation. AB - We recently found that estrogen deficiency leads to a lowering of thiol antioxidant defenses in rodent bone. Moreover, administration of agents that increase the concentration in bone of glutathione, the main intracellular antioxidant, prevented estrogen-deficiency bone loss, whereas depletion of glutathione by buthionine sulfoximine administration provoked substantial bone loss. To analyze further the mechanism by which antioxidant defenses modulate bone loss, we have now compared expression of the known antioxidant enzymes in osteoclasts. We found that glutathione peroxidase 1 (Gpx), the enzyme primarily responsible for the intracellular degradation of hydrogen peroxide, is overwhelmingly the predominant antioxidant enzyme expressed by osteoclasts and that its expression was increased in bone marrow macrophages by receptor activator of nuclear factor-kappaB ligand (RANKL) and in osteoclasts by 17beta estradiol. We therefore tested the effect of overexpression of Gpx in osteoclasts by stable transfection of RAW 264.7 (RAW) cells, which are capable of osteoclastic differentiation in response to RANKL, with a Gpx-expression construct. Osteoclast formation was abolished. The Gpx expression construct also suppressed RANKL-induced nuclear factor-kappaB activation and increased resistance to oxidation of dihydrodichlorofluorescein by exogenous hydrogen peroxide. We therefore tested the role of hydrogen peroxide in the loss of bone caused by estrogen deficiency by administering pegylated catalase to mice. We found that catalase prevented ovariectomy-induced bone loss. These results suggest that hydrogen peroxide is the reactive oxygen species responsible for signaling the bone loss of estrogen deficiency. PMID- 15528307 TI - The estrogen receptor (ER)alpha variant Delta5 exhibits dominant positive activity on ER-regulated promoters in endometrial carcinoma cells. AB - Estrogen receptor (ER)alpha is a ligand-inducible transcription factor that mediates the physiological effects of 17beta-estradiol (E2). In the uterus, E2 is involved in tissue growth, maintenance, and differentiation. Delta5ERalpha (Delta5) is an ERalpha variant protein expressed in uterine tumors but not in normal tissue. We examined the transcriptional activity of Delta5 and its modulation of human ERalpha basal and E2-stimulated activity in Ishikawa cells, an endometrial cancer cell line. In transient transfection assays, Delta5 increased basal activity of an estrogen response element-containing promoter in the absence or presence of ERalpha but lessened stimulation by ERalpha and E2. Effects of Delta5 were not limited to model reporters, given that cyclin D1 and complement 3 promoters were similarly affected. Increases in basal transcription required dimerization and DNA binding of Delta5, whereas decreased E2 stimulation with ERalpha required only DNA binding. Decreased ligand stimulation was not unique to E2 but also applied to the selective ER modulators tamoxifen and genistein. However, promoter stimulation by epidermal growth factor is retained with Delta5. The ERalpha coactivator small nuclear ring finger protein is expressed in Ishikawa cells and uterine tumors, and it enhances effects of Delta5 alone and with ERalpha on basal activity of an estrogen response element reporter. Thus, in the presence of Delta5 plus ERalpha, there is a lower transcriptional response to E2 and SERMS, but stimulation by epidermal growth factor is retained. The expression of Delta5 in uterine carcinoma may provide a mechanism by which tumors could maintain expression of E2-responsive genes in the absence of E2. PMID- 15528308 TI - Role of the duodenum and macronutrient type in ghrelin regulation. AB - The orexigenic hormone ghrelin is implicated in preprandial hunger and meal initiation in part because circulating levels increase before meals and decrease after food intake. The mechanisms underlying postprandial ghrelin suppression are unknown. Although most ghrelin is produced by the stomach, we have shown that neither gastric nutrients nor gastric distension affect ghrelin levels. We hypothesized that the nutrient-sensing mechanism regulating ghrelin is in the duodenum, the second richest source of ghrelin. To test whether duodenal nutrient exposure is required for ghrelin suppression, we infused nutrients into either the proximal duodenum or proximal jejunum in rats bearing chronic intestinal cannulas. At 0, 30, 60, 90, 120, 180, 240, and 300 min after infusions, blood was sampled via jugular-vein catheters for ghrelin, insulin, and glucose measurements. To elucidate further the mechanisms governing nutrient-related ghrelin suppression, we also assessed the ghrelin responses to isocaloric (3 kcal) infusions of glucose, amino acids, or lipids delivered into the stomach or small intestine of chronically catheterized rats. Regardless of macronutrient type, the depth and duration of ghrelin suppression were equivalent after gastric, duodenal, and jejunal infusions. Glucose and amino acids suppressed ghrelin more rapidly and strongly (by approximately 70%) than did lipids (by approximately 50%). Because jejunal nutrient infusions suppressed ghrelin levels as well as either gastric or duodenal infusions, we conclude that the inhibitory signals mediating postprandial ghrelin suppression are not derived discretely from either the stomach or duodenum. The relatively weak suppression of ghrelin by lipids compared with glucose or amino acids could represent one mechanism promoting high-fat dietary weight gain. PMID- 15528309 TI - Platelet P-selectin is required for pulmonary eosinophil and lymphocyte recruitment in a murine model of allergic inflammation. AB - Platelets are necessary for lung leukocyte recruitment in a murine model of allergic inflammation, and platelet-leukocyte aggregates are formed in circulating blood of patients with asthma after allergen exposure. However, it is unknown how platelets induce pulmonary leukocyte recruitment in asthma. Here, we have investigated the importance of platelet adhesion molecule expression on pulmonary eosinophil and lymphocyte recruitment and on leukocyte CD11b and very late antigen (VLA)-4 expression in mice. Pulmonary leukocyte recruitment in platelet-depleted mice (sensitized and exposed to ovalbumin) transfused with fixed, unstimulated platelets (FUSPs) was abolished, whereas transfusion with platelets stimulated and fixed (FSPs), expressing P-selectin and P-selectin glycoprotein ligand-1 (PSGL-1), restored eosinophil and lymphocyte recruitment. Transfusion with platelets from P-selectin-deficient mice, or with FSPs stimulated in the presence of a blocking anti-P-selectin antibody, were unable to restore pulmonary leukocyte recruitment. Flow cytometric analysis revealed increased expression of CD11b and VLA-4 on leukocytes attached to platelets after allergen exposure, and CD11b expression on leukocytes was suppressed in thrombocytopenic mice but was restored with the transfusion of FSPs, but not FUSPs, a phenomenon concurrent with the formation of platelet-leukocyte complexes. P-selectin expression on the surfaces of platelets is a major requirement for pulmonary eosinophil and lymphocyte recruitment, allowing circulating platelets to bind to and stimulate leukocytes for endothelial attachment. PMID- 15528310 TI - In vitro maturation of nascent reticulocytes to erythrocytes. AB - Most studies of mammalian reticulocyte maturation have used blood reticulocytes. Nascent reticulocytes, as found in bone marrow, have not been available in developmentally synchronized populations. Nascent murine reticulocytes formed in vitro by enucleation of Friend virus-infected erythroblasts were purified and recultured for 110 hours. At 0 hours, all recultured cells were lobulated and contained dense, centralized reticulin. By 110 hours, about 20% to 25% of the cells became biconcave erythrocytes. Most ribosomes and cellular RNAs were degraded within 20 hours, and during that period, heme synthesis declined from a rate equal to that of late erythroblasts to less than 10% of that rate. Many mitochondria appeared normal until they showed outer membrane swelling, degradation, and apparent fusion with intracellular vacuoles at 40 hours of culture. During the period of mitochondrial loss, Bcl-X(L), an antiapoptotic protein that accumulates during erythroblast differentiation and maintains mitochondrial membrane integrity, demonstrated progressive decreases and changes consistent with deamidation. Nevertheless, the reticulocytes did not undergo apoptosis, because their apoptotic machinery was degraded. This experimental system that provides a developmentally synchronized population of nascent murine reticulocytes that mature into biconcave erythrocytes in vitro should be useful in further investigations of the cellular events involved in reticulocyte maturation. PMID- 15528311 TI - Iron trafficking in the mitochondrion: novel pathways revealed by disease. AB - It is well known that iron (Fe) is transported to the mitochondrion for heme synthesis. However, only recently has the importance of this organelle for many other facets of Fe metabolism become widely appreciated. Indeed, this was stimulated by the description of human disease states that implicate mitochondrial Fe metabolism. In particular, studies assessing various diseases leading to mitochondrial Fe loading have produced intriguing findings. For instance, the disease X-linked sideroblastic anemia with ataxia (XLSA/A) is due to a mutation in the ATP-binding cassette protein B7 (ABCB7) transporter that is thought to transfer [Fe-S] clusters from the mitochondrion to the cytoplasm. This and numerous other findings suggest the mitochondrion is a dynamo of Fe metabolism, being vital not only for heme synthesis but also for playing a critical role in the genesis of [Fe-S] clusters. Studies examining the disease Friedreich ataxia have suggested that a mutation in the gene encoding frataxin leads to mitochondrial Fe loading. Apart from these findings, the recently discovered mitochondrial ferritin that may store Fe in ring sideroblasts could also regulate the level of Fe needed for heme and [Fe-S] cluster synthesis. In this review, we suggest a model of mitochondrial Fe processing that may account for the pathology observed in these disease states. PMID- 15528312 TI - A cardioprotective role for the endothelial protein C receptor in lipopolysaccharide-induced endotoxemia in the mouse. AB - A model of gram-negative lethal endotoxin shock, involving continuous peritoneal infusion of lipopolysaccharide (LPS), has been applied to wild-type (WT) mice and mice with a severe deficiency of endothelial protein C receptor (EPCR(delta/delta)). The survival of EPCR(delta/delta) mice was significantly diminished as compared to WT mice after administration of LPS via this route. Heart rates and central blood pressures also were significantly more depressed in EPCR(delta/delta) mice, indicating that the receptor-based protein C (PC) pathway functions in regulation of hemodynamic properties in the mouse. Further, heart muscle damage was more severe in EPCR(delta/delta) mice as compared to WT mice after endotoxin administration, as revealed by the more elevated plasma myoglobin levels in EPCR(delta/delta) mice and by microscopic examination of stained heart sections. Neutrophil infiltration was more pronounced in heart tissue of EPCR(delta/delta) mice, perhaps in response to the greatly increased expression level of the chemokine, MIP-2, which also significantly more up-regulated in the LPS-treated EPCR(delta/delta) mouse cohort. In conclusion, a severe deficiency of EPCR adversely affects survival of mice subjected to continuous infusion of endotoxin, via contributions of more responsive hemodynamic and cardiac alterations, thus suggesting that, among its other functions, the PC-based receptor system has a cardioprotective role after acute inflammatory challenge. PMID- 15528313 TI - Immunotherapy of autoimmune encephalomyelitis with redirected CD4+CD25+ T lymphocytes. AB - We developed an approach that increases CD4+CD25+ regulatory T-cell potency by antigen-specifically redirecting them against pathologic T lymphocytes. The regulatory cells are transgenically modified with chimeric receptors that link antigen-major histocompatibility complex (MHC) extracellular and transmembrane domains with the cytoplasmic signaling tail of T-cell receptor zeta (TCR-zeta). The receptors' antigen-MHC recognizes the TCR of cognate T lymphocytes. Receptor engagement stimulates the receptor-modified T cell (RMTC) through the linked zeta chain. CD4+CD25+ RMTCs expressing a myelin basic protein (MBP) 89-101-IAs-zeta receptor, unlike unmodified CD4+CD25+ T cells or CD4+CD25- RMTCs, prevented and treated experimental allergic encephalomyelitis (EAE) induced with MBP89-101. The RMTCs were effective even after the autoreactive T-cell repertoire had diversified to include specificities not directly targeted by the chimeric receptor. Remissions were sustained and mortality was decreased from more than 50% to 0%. These results provide proof of principal for a novel approach to enforce the interaction of regulatory and pathologic T lymphocytes, thereby facilitating the treatment of autoimmune disease. PMID- 15528314 TI - Punish the parent not the progeny. AB - Chronic myeloid leukemia (CML) is sustained by a rare population of primitive, quiescent, BCR-ABL+ cells and represents an excellent example of a malignancy in which tumor-initiating cells represent the key to disease eradication. CML is also the first malignancy for which targeted therapy has replaced conventional chemotherapy. Within a vast excess of proliferating progenitor cells that express breakpoint cluster region-abelson (BCR-ABL) and are exquisitely sensitive to the tyrosine kinase inhibitor imatinib mesylate (IM) resides a small population of quiescent leukemic cells that, despite higher levels of BCR-ABL transcripts, exhibits innate insensitivity to IM. These cells remain after IM therapy, even when apparently complete responses are achieved, and they probably explain molecular disease persistence. Although it can be argued that patients may survive for many years with low levels of leukemia still present, it is possible to achieve disease clearance at the molecular level following an allogeneic stem cell transplantation. The emergence of drug resistance with IM monotherapy also argues in favor of complete disease eradication that we believe should remain the ultimate therapeutic goal in CML. New approaches to the elimination of these primitive CML cells may thus be crucial to the development of curative strategies. PMID- 15528315 TI - Single-cell analysis of the human NK cell response to missing self and its inhibition by HLA class I. AB - Natural killer (NK) cells activate quickly in response to pathogens, tumors, and allogeneic hematopoietic cell transplants. Modulating the NK cell response are clonally distributed NK cell receptors that survey cells for change in the expression of major histocompatibility complex (MHC) class I and structurally related ligands. Here the enzyme-linked immunospot (ELISPOT) assay, intracellular cytokine staining (ICS), and short-term culture were used to quantify the response of bulk NK cell populations from human donors to HLA class I-deficient 221 cells and to 221 cells transfected with single HLA class I allotypes. NK cells in cultures containing interleukin-2 (IL-2) or IL-12 exhibited specificities of HLA class I-mediated inhibition that correlated well with those previously defined using NK cell clones in long-term culture and with the frequencies of cells expressing particular inhibitory HLA class I receptors. Culture with IL-12, but not IL-2, gave an increased frequency of cells expressing CD94: NKG2A but no change in killer immunoglobulin-like receptor (KIR) expression. For some heterozygote combinations of KIR3DL1 alleles, ICS can be used to compare the functional properties of the 2 allotypes. Thus, both the low expressing KIR3DL1*005 and the high-expressing KIR3DL1*002 gave similar inhibitory response on challenge with an HLA-B*5801 ligand. The single-cell assays developed here should facilitate future population study and clinical analysis of human NK cell regulation by MHC class I. PMID- 15528316 TI - Therapy-related acute myeloid leukemia-like MLL rearrangements are induced by etoposide in primary human CD34+ cells and remain stable after clonal expansion. AB - Rearrangements involving the MLL gene on chromosome band 11q23 are a hallmark of therapy-related acute myeloid leukemias following treatment with topoisomerase II poisons including etoposide. Therapy-related and de novo genomic translocation breakpoints cluster within a well-characterized 8.3-kb fragment of MLL. Repair of etoposide-stabilized DNA topoisomerase II covalent complexes may initiate MLL rearrangements observed in patients. We used a culture system of primary human hematopoietic CD34+ cells and inverse polymerase chain reaction to characterize the spectrum of stable genomic rearrangements promoted by etoposide exposure originating within an MLL translocation hotspot in therapy-related leukemia. Alterations to the region were observed at a readily detectable frequency in etoposide-treated cells. Illegitimate repair events after minimal repair included MLL tandem duplications and translocations, with minor populations of deletions or insertions. In stably repaired cells that proliferated for 10 to 14 days, the significant majority of illegitimate events were MLL tandem duplications, and several deletions, inversions, insertions, and translocations. Thus, etoposide promotes specific rearrangements of MLL consistent with the full spectrum of oncogenic events identified in leukemic samples. Although etoposide-initiated rearrangements are frequent, only a small subset of translocations occurs in cells that proliferate significantly. PMID- 15528317 TI - Membrane receptors are not required to deliver granzyme B during killer cell attack. AB - Granzyme B (GzmB), a serine protease of cytotoxic T lymphocytes and natural killer (NK) cells, induces apoptosis by caspase activation after crossing the plasma membrane of target cells. The mechanism of this translocation during killer cell attack, however, is not understood. Killer cells release GzmB and the membrane-disturbing perforin at the contact site after target recognition. Receptor-mediated import of glycosylated GzmB and release from endosomes were suggested, but the role of the cation-independent mannose 6-phosphate receptor was recently refuted. Using recombinant nonglycosylated GzmB, we observed binding of GzmB to cellular membranes in a cell type-dependent manner. The basis and functional impact of surface binding were clarified. GzmB binding was correlated with the surface density of heparan sulfate chains, was eliminated on treatment of target cells with heparinase III or sodium chlorate, and was completely blocked by an excess of catalytically inactive GzmB or GzmK. Although heparan sulfate-bound GzmB was taken up rapidly into intracellular lysosomal compartments, neither of the treatments had an inhibitory influence on apoptosis induced by externally added streptolysin O and GzmB or by natural killer cells. We conclude that membrane receptors for GzmB on target cells are not crucial for killer cell-mediated apoptosis. PMID- 15528318 TI - The hereditary hemochromatosis protein, HFE, lowers intracellular iron levels independently of transferrin receptor 1 in TRVb cells. AB - Hereditary hemochromatosis (HH) is an autosomal recessive disease that leads to parenchymal iron accumulation. The most common form of HH is caused by a single amino acid substitution in the HH protein, HFE, but the mechanism by which HFE regulates iron homeostasis is not known. In the absence of transferrin (Tf), HFE interacts with transferrin receptor 1 (TfR1) and the 2 proteins co-internalize, and in vitro studies have shown that HFE and Tf compete for TfR1 binding. Using a cell line lacking endogenous transferrin receptors (TRVb cells) transfected with different forms of HFE and TfR1, we demonstrate that even at low concentrations Tf competes effectively with HFE for binding to TfR1 on living cells. Transfection of TRVb cells or the derivative line TRVb1 (which stably expresses human TfR1) with HFE resulted in lower ferritin levels and decreased Fe2+ uptake. These data indicate that HFE can regulate intracellular iron storage independently of its interaction with TfR1. Earlier studies found that in HeLa cells, HFE expression lowers Tf-mediated iron uptake; here we show that HFE lowers non-Tf-bound iron in TRVb cells and add to a growing body of evidence that HFE may play different roles in different cell types. PMID- 15528319 TI - Metabolism of nicotine and cotinine by human cytochrome P450 2A13. AB - Nicotine, a major constituent of tobacco, plays a critical role in smoking addiction. In humans, nicotine is primarily metabolized to cotinine, which is further metabolized to trans-3'-hydroxycotinine. Recently, we have demonstrated that heterologously expressed human CYP2A13 is highly active in the metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a nicotine-derived carcinogen. In the present study, CYP2A13-catalyzed NNK metabolism was found to be inhibited competitively by nicotine and N'-nitrosonornicotine (NNN), suggesting that both nicotine and NNN are also substrates of CYP2A13. We have further demonstrated that human CYP2A13 is indeed an efficient enzyme in catalyzing C-oxidation of nicotine to form cotinine, with the apparent K(m) and V(max) values of 20.2 microM and 8.7 pmol/min/pmol, respectively. CYP2A13 also catalyzes the 3'-hydroxylation of cotinine to form trans-3'-hydroxycotinine, with the apparent K(m) and V(max) values of 45.2 microM and 0.7 pmol/min/pmol, respectively. The importance of CYP2A13-catalyzed nicotine and cotinine metabolism in vivo remains to be determined. PMID- 15528320 TI - Multiple transport systems mediate the hepatic uptake and biliary excretion of the metabolically stable opioid peptide [D-penicillamine2,5]enkephalin. AB - Rapid and extensive biliary excretion of [D-penicillamine2,5]enkephalin (DPDPE) in rats as the unchanged peptide suggests that multiple transport proteins may be involved in the hepatobiliary disposition of this zwitterionic peptide. Although DPDPE is a P-glycoprotein substrate, the role of other transport proteins in the hepatic clearance of DPDPE has not been established. Furthermore, the ability of various experimental approaches to quantitate the contribution of a specific hepatic uptake or excretion process when multiple transport systems are involved has not been addressed. 3H-DPDPE uptake in suspended Wistar rat hepatocytes was primarily (>95%) due to temperature-dependent transport mechanisms; similar results were obtained in suspended hepatocytes from Mrp2-deficient (TR-) rats. Pharmacokinetic modeling revealed that saturable and linear processes were involved in 3H-DPDPE uptake in hepatocytes. The use of transport modulators suggested that hepatic uptake of 3H-DPDPE was mediated by Oatp1a1, Oatp1a4, and likely Oatp1b2. Accumulation of 3H-DPDPE in sandwich-cultured (SC) hepatocytes was rapid; uptake of 3H-DPDPE in SC rat hepatocytes from control and TR- rats was similar. However, the biliary excretion index and biliary clearance decreased by 83 and 85%, respectively, in TR- SC rat hepatocytes, indicating that DPDPE is an Mrp2 substrate. Rate constants for uptake and excretion of 3H-DPDPE in SC rat hepatocytes were determined by pharmacokinetic modeling; data were consistent with basolateral excretion of 3H-DPDPE from the hepatocyte. These results demonstrate the complexities of hepatobiliary disposition when multiple transport mechanisms are involved for a given substrate and emphasize the necessity of multi-experimental approaches for the comprehensive resolution of these processes. PMID- 15528321 TI - TLR signaling: an emerging bridge from innate immunity to atherogenesis. AB - Chronic inflammation and disordered lipid metabolism represent hallmarks of atherosclerosis. Considerable evidence suggests that innate immune defense mechanisms might interact with proinflammatory pathways and contribute to development of arterial plaques. The preponderance of such evidence has been indirect clinical and epidemiologic studies, with some support from experimental animal models of atherosclerosis. However, recent data now directly implicate signaling by TLR4 in the pathogenesis of atherosclerosis, establishing a key link between atherosclerosis and defense against both foreign pathogens and endogenously generated inflammatory ligands. In this study, we briefly review these and closely related studies, highlighting areas that should provide fertile ground for future studies aimed at a more comprehensive understanding of the interplay between innate immune defense mechanisms, atherosclerosis, and related vascular disorders. PMID- 15528322 TI - Cutting edge: human eosinophils regulate T cell subset selection through indoleamine 2,3-dioxygenase. AB - Allergy involves eosinophilia and Th2 polarization. Indoleamine 2,3-dioxygenase (IDO)-catalyzed conversion of tryptophan to kynurenines (KYN) regulates T cell function. We show that human eosinophils constitutively express IDO. Eosinophils treated with IFN-gamma showed an 8-fold increase in IDO mRNA within 4 h; IL-3, IL 5, and GM-CSF had no effect on baseline IDO expression. IL-3 pretreatment of eosinophils reduced IFN-gamma-induced IDO mRNA expression below baseline. Conversely, GM-CSF, but not IL-5, resulted in a 2-fold increase in IFN-gamma induced IDO. Treatment with IL-3, IL-5, GM-CSF, or IFN-gamma alone expressed IDO enzymatic activity (the presence of KYN in supernatants 48 h postculture). CD28 cross-linking resulted in measurable KYN in culture supernatants, inhibitable by a neutralizing anti-IFN-gamma. Coculture of eosinophils with an IFN-gamma producing T cell line, but not IL-4-producing T cell clone, led to apoptosis and inhibition of CD3 or CD3/CD28-induced proliferation. Eosinophils infiltrating asthmatic lung and associated lymphoid tissue exhibited intracellular IDO immunoreactivity. Eosinophils may, therefore, maintain Th2 bias through IDO. PMID- 15528323 TI - Cutting edge: bone morphogenetic protein antagonists Drm/Gremlin and Dan interact with Slits and act as negative regulators of monocyte chemotaxis. AB - Drm/Gremlin and Dan, two homologous secreted antagonists of bone morphogenic proteins, have been shown to regulate early development, tumorigenesis, and renal pathophysiology. In this study, we report that Drm and Dan physically and functionally interact with Slit1 and Slit2 proteins. Drm binding to Slits depends on its glycosylation and is not interfered with by bone morphogenic proteins. Importantly, Drm and Dan function as inhibitors for monocyte migration induced by stromal cell-derived factor 1alpha (SDF-1alpha) or fMLP. The inhibition of SDF 1alpha-induced monocyte chemotaxis by Dan is not due to blocking the binding of SDF-1alpha to its receptor. Thus, the results identify that Drm and Dan can interact with Slit proteins and act as inhibitors of monocyte chemotaxis, demonstrating a previously unidentified biological role for these proteins. PMID- 15528324 TI - Cutting edge: immunity and IFN-gamma production during Listeria monocytogenes infection in the absence of T-bet. AB - The T-box transcription factor T-bet is an important regulator of IFN-gamma production in all cell types and is considered to be essential for the generation of CD4 Th1 T cells. IFN-gamma in turn plays a critical role in immunity to many infectious agents. In this study, we demonstrate that T-bet is not required for host resistance to primary Listeria monocytogenes (LM) infection. In the innate immune phase, control of LM replication, serum IFN-gamma, and numbers of IFN gamma-producing NK cells were similar in T-bet-deficient and control mice. In the adaptive immune phase, there was no defect in bacterial clearance or in the numbers of LM-specific IFN-gamma-producing CD8 T cells in T-bet-deficient mice and only a modest, although significant, reduction in the numbers of Th1 CD4 T cells and IFN-gamma secretion by CD4 T cells. Thus, host resistance and the generation of IFN-gamma-producing cells in response to LM infection are not substantially compromised in the absence of T-bet. PMID- 15528325 TI - Cutting edge: tumor-specific CTL are constitutively cross-armed in draining lymph nodes and transiently disseminate to mediate tumor regression following systemic CD40 activation. AB - The cross-arming of effector CTL in response to cross-presented tumor Ags is predicted to fail in the absence of CD40 stimulation. However, questions remain regarding the role of CD40 signaling and additional CD4+ T cell-derived signals in this process. To address this, we have analyzed the cross-arming of tumor specific CTL effectors in vivo in a mouse model of established tumor and tumor regression following CD40 activation. We found that tumor-specific CTL were constitutively cross-armed in tumor-draining lymph nodes during tumor growth and that systemic CD40 activation did not alter CTL cross-arming in the tumor draining lymph nodes. Rather, CD40 activation induced peripheral dissemination of tumor-specific CTL effectors that required continual CD40 stimulation to maintain peripheral CTL and tumor regression. These data indicate that CD40 activation enhances the peripheral survival of constitutively cross-armed CTL and that persistent CD4+ T cell signals are required for their long-term activity. PMID- 15528326 TI - Cutting edge: cross-presentation of cell-associated antigens to MHC class I molecule is regulated by a major transcription factor for heat shock proteins. AB - The ability for the professional APC to cross-present Ag to MHC class I from parenchymal cells is essential for priming as well as tolerance of CD8+ T cells against intracellular Ags. Since cross-presentations of non-cell-associated free Ags are inefficient, the roles of molecular chaperones or heat shock proteins (HSPs) in chaperoning Ags to APCs have been postulated. We herein genetically addressed this hypothesis using mice that were defective of heat shock factor 1 (Hsf1), a major transcription factor for HSPs. Hsf1(-/-) mice have a decreased expression of several HSPs including HSP90 and HSP70. Using multiple Ag systems, we demonstrated that cross-priming of Ag-specific CD8+ T cells was inefficient when Ag expression was restricted to Hsf1(-/-) non-APCs. Our study provides the first genetic evidence for the roles of Hsf1 in regulating cross-presentation of MHC class I-associated Ags. PMID- 15528327 TI - Replication-dependent potent IFN-alpha induction in human plasmacytoid dendritic cells by a single-stranded RNA virus. AB - Plasmacytoid dendritic cells sense viral ssRNA or its degradation products via TLR7/8 and CpG motifs within viral DNA via TLR9. Although these two endosomal pathways operate independently of viral replication, little is known about the detection of actively replicating viruses in plasmacytoid dendritic cell (PDC). Replication and transcription of the viral genome of ssRNA viruses as well as many DNA viruses lead to the formation of cytosolic dsRNA absent in noninfected cells. In this study, we used human respiratory syncytial virus (HRSV) encoding a fusion (F) protein for direct cytosolic entry. Both HRSV infection and cytosolic delivery of a 65-nt dsRNA led to potent IFN-alpha induction in PDC, but not in myeloid dendritic cells. Inactivation of HRSV by UV irradiation abrogated IFN alpha induction in PDC. The comparison of two respiratory syncytial virus (RSV) constructs carrying either the HRSV or the bovine RSV F protein revealed that F mediated cytosolic entry of RSV was absolutely required for IFN-alpha induction in PDC. HRSV-induced IFN-alpha production was independent of endosomal acidification and of protein kinase R (PKR) kinase activity, as demonstrated with chloroquine and the PKR inhibitor 2-aminopurine, respectively. In contrast, the induction of IFN-alpha by the TLR7/8 ligand R848, by the TLR9 ligand CpG-A ODN 2216, and by inactivated influenza virus (TLR7/8 dependent) was completely blocked by 2-aminopurine. IFN-alpha induction by mouse pathogenic Sendai virus was not affected in PKR- and MyD88-deficient mice, confirming that a ssRNA virus, which is able to directly enter host cells via fusion at the plasma membrane, can be detected by PDC independently of PKR, TLR7/8, and TLR9. PMID- 15528328 TI - 4-1BB and OX40 act independently to facilitate robust CD8 and CD4 recall responses. AB - Mice deficient in OX40 or 4-1BB costimulatory pathways show defects in T cell recall responses, with predominant effects on CD4 vs CD8 T cells, respectively. However, OX40L can also stimulate CD8 T cells and 4-1BBL can influence CD4 T cells, raising the possibility of redundancy between the two TNFR family costimulators. To test this possibility, we generated mice deficient in both 4 1BBL and OX40L. In an adoptive transfer model, CD4 T cells expressed 4-1BB and OX40 sequentially in response to immunization, with little or no overlap in the timing of their expression. Under the same conditions, CD8 T cells expressed 4 1BB, but no detectable OX40. Thus, in vivo expression of 4-1BB and OX40 can be temporally and spatially segregated. In the absence of OX40L, there were decreased CD4 T cells late in the primary response and no detectable secondary expansion of adoptively transferred CD4 T cells under conditions in which primary expansion was unaffected. The 4-1BBL had a minor effect on the primary response of CD4 T cells in this model, but showed larger effects on the secondary response, although 4-1BBL(-/-) mice show less impairment in CD4 secondary responses than OX40L(-/-) mice. The 4-1BBL(-/-) and double knockout mice were similarly impaired in the CD8 T cell response, whereas OX40L(-/-) and double knockout mice were similarly impaired in the CD4 T cell response to both protein Ag and influenza virus. Thus, 4-1BB and OX40 act independently and nonredundantly to facilitate robust CD4 and CD8 recall responses. PMID- 15528329 TI - Mechanism of prostaglandin (PG)E2-induced prolactin expression in human T cells: cooperation of two PGE2 receptor subtypes, E-prostanoid (EP) 3 and EP4, via calcium- and cyclic adenosine 5'-monophosphate-mediated signaling pathways. AB - We previously reported that prolactin gene expression in the T-leukemic cell line Jurkat is stimulated by PGE(2) and that cAMP acts synergistically with Ca(2+) or protein kinase C on the activation of the upstream prolactin promoter. Using the transcription inhibitor actinomycin D, we now show that PGE(2)-induced prolactin expression requires de novo prolactin mRNA synthesis and that PGE(2) does not influence prolactin mRNA stability. Furthermore, PGE(2)-induced prolactin expression was inhibited by protein kinase inhibitor fragment 14-22 and BAPTA-AM, which respectively, inhibit protein kinase A- and Ca(2+)-mediated signaling cascades. Using specific PGE(2) receptor agonists and antagonists, we show that PGE(2) induces prolactin expression through engagement of E-prostanoid (EP) 3 and EP4 receptors. We also found that PGE(2) induces an increase in intracellular cAMP concentration as well as intracellular calcium concentration via EP4 and EP3 receptors, respectively. In transient transfections, 3000 bp flanking the leukocyte prolactin promoter conferred a weak induction of the luciferase reporter gene by PGE(2) and cAMP, whereas cAMP in synergy with ionomycin strongly activated the promoter. Mutation of a C/EBP responsive element at -214 partially abolished the response of the leukocyte prolactin promoter to PGE(2), cAMP, and ionomycin plus cAMP. PMID- 15528330 TI - Eosinophil adhesion to cholinergic IMR-32 cells protects against induced neuronal apoptosis. AB - Eosinophils release a number of mediators that are potentially toxic to nerve cells. However, in a number of inflammatory conditions, such as asthma and inflammatory bowel disease, it has been shown that eosinophils localize to nerves, and this is associated with enhanced nerve activity. In in vitro studies, we have shown that eosinophil adhesion via neuronal ICAM-1 leads to activation of neuronal NF-kappaB via an ERK1/2-dependent pathway. In this study, we tested the hypothesis that eosinophil adhesion to nerves promotes neural survival by protection from inflammation-associated apoptosis. Exposure of differentiated IMR 32 cholinergic nerve cells to IL-1beta, TNF-alpha, and IFN-gamma, or culture in serum-deprived medium, induced neuronal apoptosis, as detected by annexin V staining, caspase-3 activation, and DNA laddering. Addition of human eosinophils to IMR-32 nerve cells completely prevented all these features of apoptosis. The mechanism of protection by eosinophils was by an adhesion-dependent activation of ERK1/2, which led to the induced expression of the antiapoptotic gene bfl-1. Adhesion to nerve cells did not influence the expression of the related genes bax and bad. Thus, prevention of apoptosis by eosinophils may be a mechanism by which these cells regulate neural plasticity in the peripheral nervous system. PMID- 15528331 TI - Rac2-deficient murine macrophages have selective defects in superoxide production and phagocytosis of opsonized particles. AB - The Rho family GTPase Rac is a crucial participant in numerous cellular functions and acts as a molecular switch for signal transduction. Mice deficient in hemopoietic-specific Rac2 exhibited agonist-specific defects in neutrophil functions including chemoattractant-stimulated filamentous actin polymerization and chemotaxis, and superoxide production elicited by phorbol ester, fMLP, or IgG coated particles, despite expression of the highly homologous Rac1 isoform. In this study, functional responses of Rac2-null murine macrophages were characterized to examine whether Rac2 also has nonredundant functions in this phagocytic lineage. In contrast to murine neutrophils, in which Rac1 and Rac2 are present in similar amounts, Rac1 was approximately 4-fold more abundant than Rac2 in both bone marrow-derived and peritoneal exudate macrophages, and macrophage Rac1 levels were unchanged by the absence of Rac2. Accumulation of exudate macrophages during peritoneal inflammation was reduced in rac2(-/-) mice. FcgammaR-mediated phagocytosis of IgG-coated SRBC was also significantly decreased in Rac2-null macrophages, as was NADPH oxidase activity in response to phorbol ester or FcgammaR stimulation. However, phagocytosis and oxidant production stimulated by serum-opsonized zymosan was normal in rac2(-/-) macrophages. Macrophage morphology was also similar in wild-type and Rac2-null cells, as was actin polymerization induced by FcgammaR-mediated phagocytosis or M CSF. Hence, Rac2-null macrophages have selective defects paralleling many of the observed functional defects in Rac2-null neutrophils. These results provide genetic evidence that although Rac2 is a relatively minor isoform in murine macrophages, it plays a nonoverlapping role with Rac1 to regulate host defense functions in this phagocyte lineage. PMID- 15528332 TI - Impaired receptor editing in the primary B cell repertoire of BASH-deficient mice. AB - The editing of B cell Ag receptor (BCR) through successive rearrangements of Ig genes has been considered to be a major mechanism for the central B cell tolerance, which precludes appearance of self-reactive B cells, through studies using anti-self-Ig transgenic/knock-in mouse systems. However, contribution of the receptor editing in the development of the normal B cell repertoire remains unclear. In addition, the signaling pathway directing this event is unknown. In this study, we demonstrate that receptor editing in anti-DNA Ig knock-in mice is impaired in the absence of an adaptor protein BASH (BLNK/SLP-65) that is involved in BCR signaling. Remarkably, the supposed hallmarks of receptor editing such as Iglambda chain expression, recombination sequence rearrangements at Igkappa loci, and presence of in-frame VkappaJkappa joins in the Igkappa loci inactivated by the recombination sequence rearrangements, were all diminished in BASH-deficient mice with unmanipulated Ig loci. BCR ligation-induced Iglambda gene recombination in vitro was also impaired in BASH-deficient B cells. Furthermore, the BASH deficient mice showed an excessive Ab response to a DNA carrier immunization, suggesting the presence of unedited DNA-reactive B cells in the periphery. These results not only define a signaling pathway required for receptor editing but indicate that the BCR-signaled receptor editing indeed operates in the development of normal B cell repertoire and contributes to establishing the B cell tolerance. PMID- 15528333 TI - Extract of Reishi polysaccharides induces cytokine expression via TLR4-modulated protein kinase signaling pathways. AB - We have demonstrated that an extract of Ganoderma lucidum (Reishi or Ling-Zhi) polysaccharides (EORP) exerts immunomodulating activities by stimulating the expression of inflammatory cytokines from mouse spleen cells. Interestingly, via responding to LPS in genetic variation of murine macrophage HeNC2 and GG2EE cell lines, and using TLR4 Ab blockage in human blood-derived monocytic macrophages, we have found that the TLR4, but not complement receptor type 3, is a putative receptor of EORP, mediating the consequent immunomodulating events associated with IL-1 gene expression. Based on our studies of reactive oxygen species production, polymyxin B inhibition, and protein tyrosine kinase (PTK) activity, we ruled out the possibility of LPS contamination in EORP. We have found that EORP differentially modulates the protein kinase (PK)-mediated signal transduction pathways associated with inflammatory cytokine IL-1. In human macrophages and murine macrophage J774A.1 cells, EORP was found to up-regulate IL 1 secretion and pro-IL-1 (precursor of IL-1) as well as IL-1-converting enzyme expression. Specifically, EORP rapidly stimulates PTK-mediated phosphorylation, followed by induction of PKs and activation of MAPKs: ERK, JNK, and p38. Using PK inhibitors in the kinase activity assays, Western blot analyses and IL-1 ELISA, we have extensively examined and dissected the role of individual PK in the regulation of pro-IL-1/IL-1. Our findings establish that EORP-mediated signaling pathways are involved in the pro-IL-1/IL-1 regulation: PTK/protein kinase C/MEK1/ERK and PTK/Rac1/p21-activated kinase/p38. PMID- 15528334 TI - CD63 as an activation-linked T cell costimulatory element. AB - Dendritic cells (DC) are unique in their capacity to either stimulate or regulate T cells, and receptor/ligand pairs on DC and T cells are critically involved in this process. In this study we present such a molecule, which was discovered by us when analyzing the functional effects of an anti-DC mAb. This mAb, 11C9, reacted strongly with DC, but only minimally with lymphocytes. In MLR it constantly reduced DC-induced T cell activation. Therefore, we assumed that mAb 11C9 primarily exerts its functions by binding to a DC-structure. This does not seem to be the case, however. Preincubation of DC with mAb 11C9 before adding T cells had no inhibitory effect on T cell responses. Retroviral expression cloning identified the 11C9 Ag as CD63. This lysosomal-associated membrane protein (LAMP 3), is only minimally expressed on resting T cells but can, as we show, quickly shift to the surface upon stimulation. Cross-linkage of that structure together with TCR-triggering induces strong T cell activation. CD63 on T cells thus represents an alternative target for mAb 11C9 with its binding to activated T cells rather than DC being responsible for the observed functional effects. This efficient CD63-mediated costimulation of T cells is characterized by pronounced induction of proliferation, strong IL-2 production and compared with CD28 enhanced T cell responsiveness to restimulation. Particularly in this latter quality CD63 clearly surpasses several other CD28-independent costimulatory pathways previously described. CD63 thus represents an activation-induced reinforcing element, whose triggering promotes sustained and efficient T cell activation and expansion. PMID- 15528335 TI - The role of B cell-mediated T cell costimulation in the efficacy of the T cell retargeting bispecific antibody BIS20x3. AB - In this study, we investigated the role of the naturally occurring B cell mediated T cell costimulation in the antitumor efficacy of the bispecific Ab BIS20x3. BIS20x3 has a dual specificity for both CD20 and CD3 and has previously been shown to effectively direct the lytic potential of cytolytic T cells toward malignant, CD20(+) B cells. BIS20x3 instigated T cell-B cell interaction caused a dose-dependent activation of T cells that was 30 times stronger when compared with T cell activation induced by monovalent anti-CD3 Abs. The activation of T cells by BIS20x3 and B cells appeared functional and resulted in the rapid induction of high lytic potential in freshly isolated peripheral T cells. BIS20x3 mediated T cell-B cell interaction resulted in a significant up-regulation of ICAM-1 on B cells and the activation of T cells was found to be dependent on the interaction of ICAM-1 with LFA-1 and trans-activation by the NF-kappaB pathway. Also, the lytic potential of freshly isolated T cells activated via BIS20x3 appeared to be dependent on NF-kappaB signaling in the target B cells. Interestingly, the costimulatory signaling effects described in this study appeared specifically related to the targeting against CD20 because targeting against CD19, by a CD3xCD19-directed bispecific Ab, was significantly less effective in inducing T cell activation and T cell-mediated B cell lysis. Together these results demonstrate that the malignant B cells actively contribute to their own demise upon CD20-directed bispecific Ab-mediated T cell targeting. PMID- 15528336 TI - Rescuing melanoma epitope-specific cytolytic T lymphocytes from activation induced cell death, by SP600125, an inhibitor of JNK: implications in cancer immunotherapy. AB - Activation-induced cell death (AICD) as well as programmed cell death (PCD) serve to control the expansion of activated T cells to limit untoward side effects of continued effector responses by T cells and to maintain homeostasis. AICD of T cells in tumor immunotherapy can be counterproductive particularly if the activated T cells undergo apoptotic death after the very first secondary encounter of the specific epitope. We examined the extent to which tumor epitope specific CTLs that are activated and expanded in an in vitro-matured dendritic cell-based primary stimulation protocol undergo AICD following their first secondary encounter of the cognate epitope. Using the MART-1(27-35) epitope as a prototype vaccine epitope, we also examined whether these CTLs could be rescued from AICD. Our results demonstrate that a substantial fraction of MART-1(27-35) epitope-specific primary CTLs undergo AICD upon the very first secondary encounter of the cognate epitope. The AICD in these CTLs is neither caspase dependent nor is it triggered by the extrinsic death signaling pathways (Fas, TNFR, etc.). These CTLs, interestingly, could be rescued from AICD by the JNK inhibitor, SP600125. We also found that SP600125 interferes with their IFN-gamma response but does not block their cytolytic function. The rescued CTLs, however, regain their capacity to synthesize IFN-gamma if continued in culture without the inhibitor. These observations have implications in tumor immunotherapy and in further studies for regulation of AICD in CTLs. PMID- 15528337 TI - The immunization site of cytokine-secreting tumor cell vaccines influences the trafficking of tumor-specific T lymphocytes and antitumor efficacy against regional tumors. AB - Tumor cells engineered to secrete cytokines, referred to as tumor cell vaccines, can often generate systemic antitumor immunity and, in many cases, cause tumor regression. We compared the efficacy of s.c. immunization or intrahepatic immunization of GM-CSF-expressing tumor cell vaccines on the growth of s.c. or orthotopic liver tumors. A chemically transformed hepatic epithelial cell line, GP7TB, derived from Fischer 344 rats, was used to generate tumor models and tumor cell vaccines. Our results demonstrated that two s.c. injections of an irradiated tumor cell vaccine significantly controlled the growth of s.c. tumors, but was completely ineffective against orthotopic liver tumors. Effector cell infiltration in liver tumors was markedly reduced compared with s.c. tumors. Enhanced apoptosis of some effector cells was observed in the liver tumors compared with the s.c. tumors. Furthermore, the T cells induced by s.c. immunization preferentially migrated to s.c. tumor sites, as demonstrated by adoptive transfer experiments. In contrast, intrahepatic immunization, using parental tumor cells admixed with adenoviruses carrying the GM-CSF gene, yielded significantly better therapeutic effects on the liver tumors than on the s.c. tumors. Adoptive transfer experiments further confirmed that the T cells induced by liver immunization preferentially migrated to the liver tumor sites. Our results demonstrate that distinct T cell populations are induced by different immunization routes. Thus, the homing behavior of T cells depends on the route of immunization and is an important factor determining the efficacy of immunotherapy for regional tumors. PMID- 15528338 TI - Final antigenic Melan-A peptides produced directly by the proteasomes are preferentially selected for presentation by HLA-A*0201 in melanoma cells. AB - The melanoma-associated protein Melan-A contains the immunodominant CTL epitope Melan-A(26/27-35)/HLA-A*0201 against which a high frequency of T lymphocytes has been detected in many melanoma patients. In this study we show that the in vitro degradation of a polypeptide encompassing Melan-A(26/27-35) by proteasomes produces both the final antigenic peptide and N-terminally extended intermediates. When human melanoma cells expressing the corresponding fragments were exposed to specific CTL, those expressing the minimal antigenic sequence were recognized more efficiently than those expressing the N-terminally extended intermediates. Using a tumor-reactive CTL clone, we confirmed that the recognition of melanoma cells expressing an N-terminally extended intermediate of Melan-A is inefficient. We demonstrated that the inefficient cytosolic trimming of N-terminally extended intermediates could offer a selective advantage for the preferred presentation of Melan-A peptides directly produced by the proteasomes. These results imply that both the proteasomes and postproteasomal peptidases limit the availability of antigenic peptides and that the efficiency of presentation may be affected by conditions that alter the ratio between fully and partially processed proteasomal products. PMID- 15528339 TI - Evidence of a novel IL-2/15R beta-targeted cytokine involved in homeostatic proliferation of memory CD8+ T cells. AB - The homeostasis of memory CD8+ T cells is regulated by cytokines. IL-15 is shown to promote the proliferation of memory CD8+ T cells, while IL-2 suppresses their division in vivo. This inhibitory effect of IL-2 appears to occur indirectly, through other cell populations including CD25+CD4+ T cells; however, the details of this mechanism remain unclear. In this study, we show that 1) both Ag experienced and memory phenotype CD8+ T cells divided after the depletion of IL-2 in vivo; 2) this division occurred normally and CD44(high)IL-2/15Rbeta(high) CD8+ T cells generated after IL-2 depletion in IL-15 knockout (KO) and in IL-7 depleted IL-15 KO mice; 3) surprisingly, the blockade of IL-2/15Rbeta signaling in IL-2-depleted IL-15 KO mice completely abolished the division of memory CD8+ T cells, although the only cytokines known to act through IL-2/15Rbeta are IL-2 and IL-15; and 4) the expression of IL-2/15Rbeta molecules on memory CD8+ T cells was required for their division induced by IL-2 depletion. These results demonstrate that the depletion of IL-2 in vivo induced memory CD8+ T cell division by an IL 15-independent but by an IL-2/15Rbeta-dependent mechanism, suggesting the existence of a novel IL-2/15Rbeta-utilizing cytokine that acts directly on memory CD8+ T cells to promote cell division. PMID- 15528340 TI - Mechanisms of peritoneal B-1a cells accumulation induced by murine lupus susceptibility locus Sle2. AB - The abundance of B-1a cells found in the peritoneal cavity of mice is under genetic control. The lupus-prone mouse New Zealand Black and New Zealand White (NZB x NZW)F(1) and its derivative NZM2410 are among the strains with the highest numbers of peritoneal B1-a cells. We have previously identified an NZM2410 genetic locus, Sle2, which is associated with the production of large numbers of B-1a cells. In this paper, we examined the mechanisms responsible for this phenotype by comparing congenic C57BL/6 mice with or without Sle2. Fetal livers generated more B-1a cells in B6.Sle2 mice, providing them with a greater starting number of B-1a cells early in life. Sle2-expressing B1-a cells proliferated significantly more in vivo than their B6 counterparts, and reciprocal adoptive transfers showed that this phenotype is intrinsic to Sle2 peritoneal B cells. The rate of apoptosis detected was significantly lower in B6.Sle2 peritoneal cavity B 1a cells than in B6, with or without exogenous B cell receptor cross-linking. Increased proliferation and decreased apoptosis did not affect Sle2 peritoneal B 2 cells. In addition, a significant number of peritoneal cavity B-1a cells were recovered in lethally irradiated B6.Sle2 mice reconstituted with B6.Igh(a) bone marrow, showing radiation resistance in Sle2 B-1a cells or its precursors. Finally, B6.Sle2 adult bone marrow and spleen were a significant source of peritoneal B-1a cells when transferred into B6.Rag2(-/-) mice. This suggests that peritoneal B-1a cells are replenished throughout the animal life span in B6.Sle2 mice. These results show that Sle2 regulates the size of the B-1a cell compartment at multiple developmental checkpoints. PMID- 15528341 TI - Induction of bystander suppression by feeding antigen occurs despite normal clonal expansion of the bystander T cell population. AB - The induction of bystander suppression, whereby the response against one Ag is suppressed when it is presented in the context of an Ag to which tolerance is already established, would be an important property of oral tolerance, because it would allow treatment of autoimmune and hypersensitivity responses where the initiating Ag is not known. Although bystander suppression has been described in oral tolerance, it is not known how its effects are mediated at the level of the bystander T cells. In addition, previous studies have not compared regimes in which Ag is fed in a tolerogenic or immunogenic manner, meaning that the possible effects of Ag competition have not been excluded. In this study we have used two populations of Ag-specific TCR transgenic CD4(+) T cells to examine the cellular basis of bystander suppression associated with oral tolerance in mice in vitro and in vivo. Our results show that bystander responses can be inhibited by feeding Ag and that these effects are more pronounced in mice fed protein in tolerogenic form than after feeding Ag with mucosal adjuvant. However, the expansion of the bystander-specific CD4(+) T cells is not influenced by the presence of oral tolerance. Thus, bystander suppression does not reflect clonal deletion or reduced clonal expansion of the bystander T cells, but may act by altering the functional differentiation of bystander T cells. PMID- 15528342 TI - Homeostatic proliferation of a Qa-1b-restricted T cell: a distinction between the ligands required for positive selection and for proliferation in lymphopenic hosts. AB - Naive T cells proliferate in response to self MHC molecules after transfer into lymphopenic hosts, a process that has been termed homeostatic proliferation (HP). Previous studies have demonstrated that HP is driven by low level signaling induced by interactions with the same MHC molecules responsible for positive selection in the thymus. Little is known about the homeostatic regulation of T cells specific for class Ib molecules, including Qa-1 and H2-M3, though it has been suggested that their capacity to undergo homeostatic expansion may be inherently limited. In this study, we demonstrate that naive 6C5 TCR transgenic T cells with specificity for Qa-1(b) have a capacity similar to conventional T cells to undergo HP after transfer into sublethally irradiated mice. Proliferation was largely dependent on the expression of beta(2)-microglobulin, and experiments with congenic recipients expressing Qa-1(a) instead of Qa-1(b) demonstrated that HP is specifically driven by Qa-1(b) and not through cross recognition of classical class I molecules. Thus, the same MHC molecule that mediates positive selection of 6C5 T cells is also required for HP. Homeostatic expansion, like positive selection, occurs in the absence of a Qa-1 determinant modifier, the dominant self-peptide bound to Qa-1 molecules. However, experiments with TAP(-/-) recipients demonstrate a clear distinction between the ligand requirements for thymic selection and HP. Positive selection of 6C5 T cells is dependent on TAP function, thus selection is presumably mediated by TAP-dependent peptides. By contrast, HP occurs in TAP(-/-) recipients, providing an example where the ligand requirements for HP are less stringent than for thymic selection. PMID- 15528343 TI - Negative regulation of NK cell activities by inhibitory receptor CD94/NKG2A leads to altered NK cell-induced modulation of dendritic cell functions in chronic hepatitis C virus infection. AB - NK cells are potent activators of dendritic cells (DCs), but it remains obscure how third-party cells affect the ability of NK cells to modulate DC functions. We show here that NK cells derived from healthy donors (N-NK), when cocultured with human liver epithelial cells, induced maturation as well as activation of DCs, such as increased migratory capacity as well as T cell stimulatory activity. In contrast, NK cells from chronic hepatitis C virus-infected donors (HCV-NK) were not capable of activating DCs under the same conditions. In comparison to N-NK, HCV-NK showed higher expression of CD94/NKG2A and produced IL-10 and TGFbeta when cultured with hepatic cells, most of which express HLA-E, a ligand for CD94/NKG2A. Blockade of NKG2A restored the ability of HCV-NK to activate DCs, which appeared to result from the reduced NK cell production of IL-10 and TGFbeta. The blockade also endowed HCV-NK with an ability to drive DCs to generate Th1-polarized CD4+ T cells. These findings show that NK cell modulation of DCs is regulated by third-party cells through NK receptor and its ligand interaction. Aberrant expression of NK receptors may have an impact on the magnitude and direction of DC activation of T cells under pathological conditions, such as chronic viral infection. PMID- 15528344 TI - Vasoactive intestinal peptide modulates Langerhans cell immune function. AB - Epidermal nerves lie in close proximity to Langerhans cells (LC) and are capable of releasing peptides that modulate LC function, including calcitonin gene related peptide and pituitary adenylate cyclase-activating polypeptide. The neuropeptide vasoactive intestinal peptide (VIP) has also been found in cutaneous nerves and mRNA, for the VIP receptor vasoactive intestinal peptide receptor type 1, and vasoactive intestinal peptide receptor type 2 have been found in murine LC and the LC-like cell line XS106. We examined the effects of VIP on LC function and cutaneous immunity. VIP inhibited elicitation of a delayed-type hypersensitivity response in previously immunized mice by epidermal cells enriched for LC content pulsed with Ag in vitro. VIP also inhibited the ability of unseparated epidermal cells to present Ag to a T cell clone and hybridoma and the ability of highly enriched LCs to present to the T cell clone. Inhibition of presentation to the hybridoma was observed with an antigenic peptide that does not require processing, suggesting that VIP is active at a step independent of Ag processing. To elucidate the mechanism(s) by which VIP may mediate these effects, we determined the effects of VIP on LC cytokine production using the XS106 cell line as a surrogate for LC. VIP augmented the production of the IL-10 in LPS stimulated XS106 cells while down-regulating IL-12 and IL-1beta production. Thus, VIP, like pituitary adenylate cyclase-activating polypeptide and calcitonin gene related peptide, down-regulates LC function and the associated immune response. PMID- 15528345 TI - Antigen targeting to CD11b allows efficient presentation of CD4+ and CD8+ T cell epitopes and in vivo Th1-polarized T cell priming. AB - Bordetella pertussis adenylate cyclase (CyaA) is an invasive bacterial toxin that delivers its N-terminal catalytic domain into the cytosol of eukaryotic cells bearing the alpha(M)beta(2) integrin (CD11b/CD18), such as myeloid dendritic cells. This allows use of engineered CyaA for targeted delivery of CD8(+) T cell epitopes into the MHC class I pathway of APC and induction of robust and protective cytotoxic responses. In this study, we demonstrate that CyaA can efficiently codeliver both a CD8(+) T cell epitope (OVA(257-264)) and a CD4(+) T cell epitope (MalE(100-114)) into, respectively, the conventional cytosolic or endocytic routes of processing of murine bone marrow-derived dendritic cells. Upon CyaA delivery, a strong potentiation of the MalE(100-114) CD4(+) T cell epitope presentation is observed as compared with the MalE protein, which depends on CyaA interaction with its CD11b receptor and its subsequent clathrin-mediated endocytosis. In vivo, CyaA induces strong and specific Th1 CD4(+) and CD8(+) T cell responses against, respectively, the MalE(100-114) and OVA(257-264) epitopes. These results underscore the potency of CyaA for design of new vaccines. PMID- 15528346 TI - Augmentation of T cell levels and responses induced by androgen deprivation. AB - Androgen has been implicated as a negative regulator of host immune function and a factor contributing to the gender dimorphism of autoimmunity. Conversely, androgen deprivation has been suggested to potentiate male host immunity. Studies have shown that removal of androgen in postpubertal male mice produces an increase in size and cellularity of primary and peripheral lymphoid organs, and enhances a variety of immune responses. Yet, few details are known about the effect of androgen removal on T cell-mediated immunity. In this study, we demonstrate two pronounced and independent alterations in T cell immunity that occur in response to androgen deprivation, provided by castration, in postpubertal male mice. First, we show that levels of T cells in peripheral lymphoid tissues of mice are increased by androgen deprivation. Second, T cells from these mice transiently proliferate more vigorously to TCR- and CD28-mediated costimulation as well as to Ag-specific activation. In addition, androgen deprivation accelerates normalization of host T and B cell levels following chemotherapy-induced lymphocyte depletion. Such alterations induced by androgen deprivation may have implications for enhancing immune responses to immunotherapy and for accelerating the recovery of the immune system following chemotherapy. PMID- 15528347 TI - A general survey of thymocyte differentiation by transcriptional analysis of knockout mouse models. AB - The thymus is the primary site of T cell lymphopoiesis. To undergo proper differentiation, developing T cells follow a well-ordered genetic program that strictly depends on the heterogeneous and highly specialized thymic microenvironment. In this study, we used microarray technology to extensively describe transcriptional events regulating alphabeta T cell fate. To get an integrated view of these processes, both whole thymi from genetically engineered mice together with purified thymocytes were analyzed. Using mice exhibiting various transcriptional perturbations and developmental blockades, we performed a transcriptional microdissection of the organ. Multiple signatures covering both cortical and medullary stroma as well as various thymocyte maturation intermediates were clearly defined. Beyond the definition of histological and functional signatures (proliferation, rearrangement), we provide the first evidence that such an approach may also highlight the complex cross-talk events that occur between maturing T cells and stroma. Our data constitute a useful integrated resource describing the main gene networks set up during thymocyte development and a first step toward a more systematic transcriptional analysis of genetically modified mice. PMID- 15528348 TI - Regulated recruitment of MHC class II and costimulatory molecules to lipid rafts in dendritic cells. AB - T cell activation has long been associated with the partitioning of Ag receptors and associated molecules to lipid microdomains. We now show that dendritic cells (DCs) also accomplish the selective recruitment to lipid rafts of molecules critical for Ag presentation. Using mouse bone marrow-derived DCs, we demonstrate that MHC class II molecules become substantially localized to rafts upon DC maturation. Even more striking is the fact that CD86 is recruited to rafts upon T cell-DC interaction. Recruitment is Ag dependent and requires CD28 on T cells. Despite the regulated recruitment of MHC class II and CD86 to rafts, unlike the counter-receptors in T cells, DCs do not polarize these molecules to sites of DC T cell contact. This difference may reflect the necessity for DCs to interact with multiple T cells simultaneously and emphasizes that the biochemical and morphological correlates of lipid rafts are not necessarily equivalent. PMID- 15528349 TI - Expression and characterization of a novel CD6 ligand in cells derived from joint and epithelial tissues. AB - CD6 is a T cell surface glycoprotein that plays an important role in interactions of thymocytes with thymic epithelial cells and in mature T cell interactions with selected nonprofessional tissue APCs. We describe a novel CD6 ligand (CD6L) 3A11 Ag that is distinct from the known CD6L (CD166). The 3A11 protein is expressed on cells derived from human thymus, skin, synovium, and cartilage, and its expression is enhanced by IFN-gamma. mAbs directed against the 3A11 Ag and CD166 exhibit distinct patterns of binding to a panel of cell lines. Confocal microscopy shows that both CD166 and the 3A11 Ag are expressed at the cell surface, and that these proteins colocalize. The 3A11 Ag has a molecular mass of 130 kDa and is immunoprecipitated using either mAb 3A11 or soluble CD6-Ig fusion protein. mAbs directed against individual CD6L were less potent than was soluble CD6-Ig fusion protein in reducing adhesion of T cells to adherent 3A11-positive epithelial cells in vitro, suggesting that these Abs recognize epitopes on the 3A11 Ag and CD166 that are distinct from CD6 binding sites. Finally, transfection of epithelial cells with CD166-specific small interfering RNAs significantly decreased CD166 expression without alteration in 3A11 Ag levels, and thus confirmed that these two CD6L are distinct. Taken together, our data identifies a novel 130-kDa CD6L that may mediate interactions of synovial and epithelial cells with T lymphocytes. PMID- 15528351 TI - Vesicle size influences the trafficking, processing, and presentation of antigens in lipid vesicles. AB - Although it is accepted that particulate Ags are more immunogenic than soluble Ags in vivo, it is unclear whether this effect can be explained solely through enhanced uptake by APCs. In this study we demonstrate that vesicle size modulated the efficiency of Ag presentation by murine macrophages and that this effect was accompanied by a profound change in trafficking of Ag. Ag prepared in large particles (560 nm) was delivered into early endosome-like, immature phagosomes, whereas smaller vesicles (155 nm) and soluble Ags localized rapidly to late endosomes/lysosomes. However, peptide/class II complexes could be detected in both compartments. Phagosomes formed on uptake of large vesicles recruit Ag processing apparatus while retaining the characteristics of early endosomes. In contrast, smaller vesicles bypassed this compartment, appeared to go more rapidly to lysosomal compartments, and exhibited reduced Ag-presenting efficiency. We conclude that the ability of phagocytosed, particulate Ag to target early phagosomes results in more efficient Ag presentation. PMID- 15528350 TI - Human ribonuclease A superfamily members, eosinophil-derived neurotoxin and pancreatic ribonuclease, induce dendritic cell maturation and activation. AB - A number of mammalian antimicrobial proteins produced by neutrophils and cells of epithelial origin have chemotactic and activating effects on host cells, including cells of the immune system. Eosinophil granules contain an antimicrobial protein known as eosinophil-derived neurotoxin (EDN), which belongs to the RNase A superfamily. EDN has antiviral and chemotactic activities in vitro. In this study, we show that EDN, and to a lesser extent human pancreatic RNase (hPR), another RNase A superfamily member, activates human dendritic cells (DCs), leading to the production of a variety of inflammatory cytokines, chemokines, growth factors, and soluble receptors. Human angiogenin, a RNase evolutionarily more distant to EDN and hPR, did not display such activating effects. Additionally, EDN and hPR also induced phenotypic and functional maturation DCs. These RNases were as efficacious as TNF-alpha, but induced a different set of cytokine mediators. Furthermore, EDN production by human macrophages could be induced by proinflammatory stimuli. The results reveal the DC-activating activity of EDN and hPR and suggest that they are likely participants of inflammatory and immune responses. A number of endogenous mediators in addition to EDN have been reported to have both chemotactic and activating effects on APCs, and can thus amplify innate and Ag-specific immune responses to danger signals. We therefore propose these mediators be considered as endogenous multifunctional immune alarmins. PMID- 15528352 TI - Dietary docosahexaenoic acid suppresses T cell protein kinase C theta lipid raft recruitment and IL-2 production. AB - To date, the proximal molecular targets through which dietary n-3 polyunsaturated fatty acids (PUFA) suppress the inflammatory process have not been elucidated. Because cholesterol and sphingolipid-enriched rafts have been proposed as platforms for compartmentalizing dynamically regulated signaling assemblies at the plasma membrane, we determined the in vivo effects of fish oil and highly purified docosahexaenoic acid (DHA; 22:6n-3) on T cell microdomain lipid composition and the membrane subdomain distribution of signal-transducing molecules (protein kinase C (PKC)theta;, linker for activation of T cells, and Fas/CD95), before and after stimulation. Mice were fed diets containing 5 g/100 g corn oil (control), 4 g/100 g fish oil (contains a mixture of n-3 PUFA) plus 1 g/100 g corn oil, or 4 g/100 g corn oil plus 1 g/100 g DHA ethyl ester for 14 days. Dietary n-3 PUFA were incorporated into splenic T cell lipid raft and soluble membrane phospholipids, resulting in a 30% reduction in raft sphingomyelin content. In addition, polyclonal activation-induced colocalization of PKCtheta; with lipid rafts was reduced by n-3 PUFA feeding. With respect to PKCtheta; effector pathway signaling, both AP-1 and NF-kappaB activation, IL-2 secretion, and lymphoproliferation were inhibited by fish oil feeding. Similar results were obtained when purified DHA was fed. These data demonstrate for the first time that dietary DHA alters T cell membrane microdomain composition and suppresses the PKCtheta; signaling axis. PMID- 15528353 TI - I kappa B kinase complex alpha kinase activity controls chemokine and high endothelial venule gene expression in lymph nodes and nasal-associated lymphoid tissue. AB - The lymphotoxin (LT) beta receptor plays a critical role in secondary lymphoid organogenesis and the classical and alternative NF-kappaB pathways have been implicated in this process. IKKalpha is a key molecule for the activation of the alternative NF-kappaB pathway. However, its precise role and target genes in secondary lymphoid organogenesis remain unknown, particularly with regard to high endothelial venules (HEV). In this study, we show that IKKalpha(AA) mutant mice, who lack inducible kinase activity, have hypocellular lymph nodes (LN) and nasal associated lymphoid (NALT) tissue characterized by marked defects in microarchitecture and HEV. In addition, IKKalpha(AA) LNs showed reduced lymphoid chemokine CCL19, CCL21, and CXCL13 expression. IKKalpha(AA) LN- and NALT-HEV were abnormal in appearance with reduced expression of peripheral node addressin (PNAd) explained by a severe reduction in the HEV-associated proteins, glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1), and high endothelial cell sulfotransferase, a PNAd-generating enzyme that is a target of LTalphabeta. In this study, analysis of LTbeta(-/-) mice identifies GlyCAM-1 as another LTbeta dependent gene. In contrast, TNFRI(-/-) mice, which lose classical NF-kappaB pathway activity but retain alternative NF-kappaB pathway activity, showed relatively normal GlyCAM-1 and HEC-6ST expression in LN-HEV. In addition, in this communication, it is demonstrated that LTbetaR is prominently expressed on LN- and NALT-HEV. Thus, these data reveal a critical role for IKKalpha in LN and NALT development, identify GlyCAM-1 and high endothelial cell sulfotransferase as new IKKalpha-dependent target genes, and suggest that LTbetaR signaling on HEV can regulate HEV-specific gene expression. PMID- 15528354 TI - Follicular dendritic cell regulation of CXCR4-mediated germinal center CD4 T cell migration. AB - Follicular dendritic cells (FDCs) up-regulate the chemokine receptor CXCR4 on CD4 T cells, and a major subpopulation of germinal center (GC) T cells (CD4(+)CD57(+)), which are adjacent to FDCs in vivo, expresses high levels of CXCR4. We therefore reasoned that GC T cells would actively migrate to stromal cell-derived factor-1 (CXCL12), the CXCR4 ligand, and tested this using Transwell migration assays with GC T cells and other CD4 T cells (CD57(-)) that expressed much lower levels of CXCR4. Unexpectedly, GC T cells were virtually nonresponsive to CXCL12, whereas CD57(-)CD4 T cells migrated efficiently despite reduced CXCR4 expression. In contrast, GC T cells efficiently migrated to B cell chemoattractant-1/CXCL13 and FDC supernatant, which contained CXCL13 produced by FDCs. Importantly, GC T cell nonresponsiveness to CXCL12 correlated with high ex vivo expression of regulator of G protein signaling (RGS), RGS13 and RGS16, mRNA and expression of protein in vivo. Furthermore, FDCs up-regulated both RGS13 and RGS16 mRNA expression in non-GC T cells, resulting in their impaired migration to CXCL12. Finally, GC T cells down-regulated RGS13 and RGS16 expression in the absence of FDCs and regained migratory competence to CXCL12. Although GC T cells express high levels of CXCR4, signaling through this receptor appears to be specifically inhibited by FDC-mediated expression of RGS13 and RGS16. Thus, FDCs appear to directly affect GC T cell migration within lymphoid follicles. PMID- 15528355 TI - B cells expressing Bcl-2 and a signaling-impaired BAFF-specific receptor fail to mature and are deficient in the formation of lymphoid follicles and germinal centers. AB - The TNF family cytokine B cell-activating factor belonging to the TNF family (BAFF) (BLyS) plays a fundamental role in regulating peripheral B cell survival and homeostasis. A BAFF-specific receptor (BAFF-R; BR3) appears to mediate these functions via activation of the NF-kappaB2 pathway. Signaling by the BAFF-R is also required to sustain the germinal center (GC) reaction. Engagement of this receptor results in the induction of Bcl-2, suggesting that this antiapoptotic factor acts downstream of the BAFF-R and NF-kappaB2 pathway to promote peripheral B cell survival during primary and Ag-driven development. To test this idea, we created lines of mice coexpressing a Bcl-2 transgene and a signaling-deficient form of the BAFF-R derived from the B lymphopenic A/WySnJ strain. Surprisingly, although dramatically elevated numbers of B cells accumulate in the periphery of these mice, these B cells exhibit extremely perturbed primary development, formation of lymphoid microenvironments, and GC and IgG responses. Moreover, mice expressing the bcl-2 transgene alone display a loss of marginal zone B cells, an expansion of follicular B cells that appear immature, and alterations of the GC reaction. These results suggest that the BAFF-R and Bcl-2 regulate key and nonoverlapping aspects of peripheral B cell survival and development. PMID- 15528356 TI - PIASy-deficient mice display modest defects in IFN and Wnt signaling. AB - Protein inhibitors of activated STATs (PIAS) represent a small family of nuclear proteins that modulate the activity of many transcription factors and act as E3 ligases for covalent modification of proteins with the small ubiquitin-like modifier (SUMO). In particular, PIASy has been shown to inhibit the activation of gene expression by the IFN-responsive transcription factor STAT1 and the Wnt responsive transcription factor LEF1. To assess the function of PIASy in vivo, we generated and analyzed mice carrying a targeted mutation of the Piasy gene. We find that homozygous mutant mice have no obvious morphological defects and have a normal distribution of lymphocyte populations. Molecular analysis of signaling in response to IFN-gamma and Wnt agonists revealed a modest reduction in the activation of endogenous and transfected target genes. Two-dimensional analysis of total proteins and SUMO-modified proteins in transformed pre-B cells showed no significant differences between wild-type mice and homozygous mutant mice. Taken together, our data indicate that PIASy has a modest effect on cytokine and Wnt signaling, suggesting a redundancy with other members of the family of PIAS proteins. PMID- 15528357 TI - Long distance control of MHC class II expression by multiple distal enhancers regulated by regulatory factor X complex and CIITA. AB - MHC class II (MHC-II) genes are regulated by an enhanceosome complex containing two gene-specific transcription factors, regulatory factor X complex (RFX) and CIITA. These factors assemble on a strictly conserved regulatory module (S-X-X2 Y) found immediately upstream of the promoters of all classical and nonclassical MHC-II genes as well as the invariant chain (Ii) gene. To identify new targets of RFX and CIITA, we developed a computational approach based on the unique and highly constrained architecture of the composite S-Y motif. We identified six novel S'-Y' modules situated far away from the promoters of known human RFX- and CIITA-controlled genes. Four are situated at strategic positions within the MHC II locus, and two are found within the Ii gene. These S'-Y' modules function as transcriptional enhancers, are bona fide targets of RFX and CIITA in B cells and IFN-gamma-induced cells, and induce broad domains of histone hyperacetylation. These results reveal a hitherto unexpected level of complexity involving long distance control of MHC-II expression by multiple distal regulatory elements. PMID- 15528358 TI - Differential gene expression modulated by the cytoplasmic domain of Fc gamma RIa (CD64) alpha-chain. AB - The cytoplasmic domain (CY) of the ligand-binding alpha-chain of the gamma-chain associated FcRs can modulate receptor function such as phagocytosis, endocytosis, and intracellular trafficking of receptor-Ag complexes. To assess the potential role of the CY domain of human FcgammaRIa (CD64) alpha-chain in the transcriptional regulation of receptor-induced gene expression, we developed stably transfected murine macrophage cell lines expressing a full-length or a CY deletion mutant (tail-less) of human FcgammaRIa to analyze gene expression in response to receptor-specific cross-linking. Using the Affymetrix murine genome U74Av2 GeneChip array, we observed >100 candidate genes having > or =2-fold difference expression at 1.5 and 3 h after stimulation. Focusing on several immunologically related genes, we confirmed differential expression of M-CSF, macrophage inhibitory cytokine-1, leukocyte-specific protein 1, MIP-2, and IL-1R antagonist by RT-PCR and RNase protection assays. Analysis of mRNA stability indicated that the differential regulation of gene expression by the CY of the CD64 alpha-chain is at the level of gene transcription. Our results indicate that the CY of the CD64 alpha-chain modulates transcriptional activity induced by receptor-specific engagement in macrophages and provides a framework for understanding distinct expression profiles elicited by different Fc gamma-chain associated receptors. PMID- 15528359 TI - The Bax N terminus is required for negative regulation by the mitogen-activated protein kinase kinase and Akt signaling pathways in T cells. AB - The Bcl-2 family proapoptotic protein, Bax, redistributes to the mitochondrion in response to varied stimuli, triggering loss of mitochondrial integrity and apoptosis. Suppression of MAPK kinase (MEK1) by the reagent UO126 in activated T cells maintained in the cytokine IL-2 disrupts cytoplasmic localization of Bax and cell survival. UO126 triggers mitochondrial translocation of ectopically expressed Bax-GFP, and both UO126 and dominant negative MEK-1 (DN-MEK1) trigger increased apoptosis in Bax-GFP-expressing T cell lines. Because inhibition of PI3K or its target Akt also triggers mitochondrial translocation of Bax in T cells and apoptosis in Bax-transfected cell lines, we generated Bax deletion mutants to identify the region(s) that confers sensitivity to regulation by MEK1 and Akt. A deletion mutant (Bax(1-171)) without the C terminus mitochondrial targeting sequence or an Akt target site (Ser(184)) localizes to the cytoplasm and triggers low level apoptosis that is enhanced by DN-Akt or DN-MEK1. A construct that lacks the first 29 aa (Bax-delta29) largely localizes to mitochondria, is highly apoptogenic, and is not inhibited by Akt or MEK1. Furthermore, Bax-delta29 overcomes IL-2-dependent survival in a T cell line, whereas Bax triggers comparatively low levels of apoptosis in these cells. Cytoplasmic localization and regulation by MEK1 and Akt are restored in a mutant deleted of the first 13 aa (Bax-delta13). Taken together, our results identify a region in the Bax N terminus that determines cellular localization regulated by MEK- and Akt-dependent signaling in T cells. PMID- 15528360 TI - HIV-1 Tat-mediated effects on focal adhesion assembly and permeability in brain microvascular endothelial cells. AB - The blood-brain barrier (BBB) is a network formed mainly by brain microvascular endothelial cells (BMECs). The integrity of the BBB is critical for brain function. Breakdown of the BBB is commonly seen in AIDS patients with HIV-1 associated dementia despite the lack of productive HIV infection of the brain endothelium. The processes by which HIV causes these pathological conditions are not well understood. In this study we characterized the molecular mechanisms by which Tat mediates its pathogenic effects in vitro on primary human BMECs (HBMECs). Tat treatment of HBMECs stimulated cytoskeletal organization and increased focal adhesion sites compared with control cells or cells treated with heat-inactivated Tat. Pretreatment with Tat Abs or with the specific inhibitor SU 1498, which interferes with vascular endothelial growth factor receptor type 2 (Flk-1/KDR) phosphorylation, blocked the ability of Tat to stimulate focal adhesion assembly and the migration of HBMECs. Focal adhesion kinase (FAK) was tyrosine-phosphorylated by Tat and was found to be an important component of focal adhesion sites. Inhibition of FAK by the dominant interfering mutant form, FAK-related nonkinase, significantly blocked HBMEC migration and disrupted focal adhesions upon Tat activation. Furthermore, HIV-Tat induced permeability changes in HBMECs in a time-dependent manner. Tat also impaired BBB permeability, as observed in HIV-1 Tat transgenic mice. These studies define a mechanism for HIV-1 Tat in focal adhesion complex assembly in HBMECs via activation of FAK, leading to cytoskeletal reorganization and permeability changes. PMID- 15528361 TI - Identification and partial characterization of a variant of human CXCR3 generated by posttranscriptional exon skipping. AB - Chemokines are recognized as functionally important in many pathological disorders, which has led to increased interest in mechanisms related to the regulation of chemokine receptor (CKR) expression. Known mechanisms for regulating CKR activity are changes in gene expression or posttranslational modifications. However, little is known about CKR with respect to a third regulatory mechanism, which is observed among other seven-transmembrane receptor subfamilies, the concept of differential splicing or processing of heteronuclear RNA. We now report on the discovery of a variant human CKR, CXCR3, resulting from alternative splicing via exon skipping. The observed RNA processing entails a drastically altered C-terminal protein sequence with a predicted four- or five transmembrane domain structure, differing from all known functional CKR. However, our data indicate that that this splice variant, which we termed CXCR3-alt, despite its severe structural changes still localizes to the cell surface and mediates functional activity of CXCL11. PMID- 15528362 TI - Enhancer role of STAT5 in CD2 activation of IFN-gamma gene expression. AB - IFN-gamma is an important immunoregulatory protein with tightly controlled expression in activated T and NK cells. Three potential STAT binding regions have been recognized within the IFN-gamma promoter: 1) an IL-12-mediated STAT4 binding site at -236 bp; 2) a newly identified IL-2-induced STAT5 binding element at -3.6 kb; and 3) CD2-mediated STAT1 and STAT4 binding to an intronic element in mucosal T cells. However, functional activation of these sites remains unclear. In this study we demonstrate CD2-mediated activation of the newly characterized -3.6-kb IFN-gamma STAT5 binding region. CD2 signaling of human PBMC results in activation of the -3.6-kb IFN-gamma promoter, whereas mutation of the -3.6-kb STAT5 site attenuates promoter activity. Functional activation is accompanied by STAT5A but little STAT5B nucleoprotein binding to the IFN-gamma STAT5 site, as determined by competition and supershift assays. STAT5 activation via CD2 occurs independent of IL-2. Western and FACS analysis shows increased phospho-STAT5 following CD2 signaling. AG490, a tyrosine kinase inhibitor affecting Jak proteins, inhibits CD2-mediated IFN-gamma mRNA expression, secretion, and nucleoprotein binding to the IFN-gamma STAT5 site in a dose-dependent fashion. This report is the first to describe CD2-mediated activation of STAT5 and supports STAT5 involvement in regulation of IFN-gamma expression. PMID- 15528363 TI - The 3' untranslated region of the membrane-bound IL-1R accessory protein mRNA confers tissue-specific destabilization. AB - IL-1alpha and IL-1beta are proinflammatory cytokines that promote activation of intracellular signaling cascades, leading to stabilization of certain mRNAs and activation of transcription factors. IL-1R type I (IL-1RI) binds IL-1alpha and IL 1beta, and subsequent recruitment of the membrane-bound IL-1R accessory protein (mIL-1RAcP) facilitates signal transduction. Two alternatively spliced isoforms, soluble IL-1RAcP (sIL-1RAcP) and sIL-1RAcP-beta, which lack transmembrane and intracellular domains, have been described. The sIL-1RAcP and possibly sIL-1RAcP beta can inhibit IL-1 signaling. Proportional expression of the different IL 1RAcP splice variants may be an important determinant of responsiveness to IL-1. We show that although both mIL-1RAcP and sIL-1RAcP mRNAs are widely expressed in human tissue, their relative proportions differ significantly in a tissue specific manner. Turnover studies revealed that the sIL-1RAcP mRNA has a half life of approximately 48 h in both the kidney cell line 293 and the hepatoma cell line HepG2. The mIL-1RAcP mRNA has a similar half-life in 293 cells, but a considerably shorter half-life of approximately 5 h in HepG2 cells. Using luciferase reporter constructs, we demonstrated that this specific destabilization of the mIL-1RAcP mRNA in the latter cell type is mediated by its 2.8-kb 3'-untranslated region. Deletion analysis further established that the cell line-specific instability does not involve AU-rich elements, but is mediated by several novel elements that appear to act independently; such elements may be recognized by proteins expressed specifically in some, but not all, tissues. These data demonstrate that the cellular capacity to respond to IL-1 is tightly regulated in a tissue-specific manner. PMID- 15528364 TI - The functional interaction of the beta 2 integrin lymphocyte function-associated antigen-1 with junctional adhesion molecule-A is mediated by the I domain. AB - Binding of the beta(2) integrin LFA-1 (alpha(L)beta(2)) to junctional adhesion molecule-A (JAM-A) has been shown to enhance leukocyte adhesion and transendothelial migration. This is mediated by the membrane-proximal Ig-like domain 2 of JAM-A; however, the location of the JAM-A binding site in LFA-1 has not been identified. We have deleted the I domain in the alpha(L) subunit of LFA 1 and expressed this alpha(L) mutant in alpha(l)-deficient Jurkat J-beta(2).7 cells to demonstrate that the I domain of LFA-1 is crucial for their adhesion to immobilized JAM-A. This was substantiated by blocking the stimulated adhesion of wild-type Jurkat T cells or monocytic Mono Mac 6 cells to JAM-A using the I domain-directed mAb TS1/22 or the small molecule antagonist BIRT 377, which stabilizes the low-affinity conformation of the I domain. The immobilized LFA-1 I domain locked in the open high-affinity conformation was sufficient to support binding of transfected Chinese hamster ovary cells expressing JAM-A. Solid-phase binding assays confirmed a direct interaction of recombinant JAM-A with the immobilized locked-open I domain. These data provide the first evidence that the I domain of LFA-1 contains a functional binding site for JAM-A. PMID- 15528365 TI - Differential regulation of primary and secondary CD8+ T cells in the central nervous system. AB - T cell accumulation and effector function following CNS infection is limited by a paucity of Ag presentation and inhibitory factors characteristic of the CNS environment. Differential susceptibilities of primary and recall CD8+ T cell responses to the inhibitory CNS environment were monitored in naive and CD8+ T cell-immune mice challenged with a neurotropic coronavirus. Accelerated virus clearance and limited spread in immunized mice was associated with a rapid and increased CNS influx of virus-specific secondary CD8+ T cells. CNS-derived secondary CD8+ T cells exhibited increased cytolytic activity and IFN-gamma expression per cell compared with primary CD8+ T cells. However, both Ag-specific primary and secondary CD8+ T cells demonstrated similar contraction rates. Thus, CNS persistence of increased numbers of secondary CD8+ T cells reflected differences in the initial pool size during peak inflammation rather than enhanced survival. Unlike primary CD8+ T cells, persisting secondary CD8+ T cells retained ex vivo cytolytic activity and expressed high levels of IFN-gamma following Ag stimulation. However, both primary and secondary CD8+ T cells exhibited reduced capacity to produce TNF-alpha, differentiating them from effector memory T cells. Activation of primary and secondary CD8+ T cells in the same host using adoptive transfers confirmed similar survival, but enhanced and prolonged effector function of secondary CD8+ T cells in the CNS. These data suggest that an instructional program intrinsic to T cell differentiation, rather than Ag load or factors in the inflamed CNS, prominently regulate CD8+ T cell function. PMID- 15528366 TI - Involvement of Fc gamma RI (CD64) in the mechanism of HIV-1 inhibition by polyclonal IgG purified from infected patients in cultured monocyte-derived macrophages. AB - The aim of this study was to investigate the mechanism of HIV-1 neutralization using monocyte-derived macrophages (MDM) in comparison to PBMC as target cells. For this purpose, we analyzed neutralizing activities of different human polyclonal IgG samples purified from sera of HIV-1-infected individuals using a single cycle infection assay. We found an increase of the neutralizing titer when macrophages vs PBMC were used as target cells. Moreover, polyclonal IgG from HIV 1-infected patients that are not able to neutralize virus when PBMC are used as target cells strongly inhibit MDM infection. Similar results were obtained with neutralizing mAbs. To explore the participation of FcgammaRs in HIV-1 inhibition, F(ab')(2) and Fab of these Igs were produced. Results indicated that both F(ab')(2) and Fab are less effective to inhibit virus replication in MDM. Moreover, competition experiments with Fc fragments of IgG from healthy donors or with purified monoclonal anti-human FcgammaRs Ab strengthen the participation of the FcgammaRs, and in particular of FcgammaRI (CD64) in HIV-1 inhibition on MDM. Mechanisms by which HIV-specific IgG inhibit virus replication in cultured macrophages are proposed and the benefit of inducing such Abs by vaccination is discussed. PMID- 15528367 TI - Single-epitope DNA vaccination prevents exhaustion and facilitates a broad antiviral CD8+ T cell response during chronic viral infection. AB - Induction of a monospecific antiviral CD8+ T cell response may pose a risk to the host due to the narrow T cell response induced. At the individual level, this may result in selection of CD8+ T cell escape variants, particularly during chronic viral infection. Second, prior immunization toward a single dominant epitope may suppress the response to other viral epitopes, and this may lead to increased susceptibility to reinfection with escape variants circulating in the host population. To address these issues, we induced a memory response consisting solely of monospecific, CD8+ T cells by use of DNA vaccines encoding immunodominant epitopes of lymphocytic choriomeningitis virus (LCMV). We analyzed the spectrum of the CD8+ T cell response and the susceptibility to infection in H 2(b) and H-2(d) mice. Priming for a monospecific, CD8+ T cell response did not render mice susceptible to viral variants. Thus, vaccinated mice were protected against chronic infection with LCMV, and no evidence indicating biologically relevant viral escape was obtained. In parallel, a broad and sustained CD8+ T cell response was generated upon infection, and in H-2(d) mice epitope spreading was observed. Even after acute LCMV infection, DNA vaccination did not significantly impair naturally induced immunity. Thus, the response to the other immunogenic epitopes was not dramatically suppressed in DNA-immunized mice undergoing normal immunizing infection, and the majority of mice were protected against rechallenge with escape variants. These findings underscore that a monospecific vaccine may induce efficient protective immunity given the right set of circumstances. PMID- 15528368 TI - Maximum immunobioactivity of murine small intestinal intraepithelial lymphocytes resides in a subpopulation of CD43+ T cells. AB - CD43 has been linked to many function-associated T cell activities. Using mAbs that recognize two different CD43 determinants, we show that, although mouse small intestinal intraepithelial lymphocytes (IELs) expressed the CD43 core molecule reactive with mAb R2/60, only about one-half of the total IELs-including some but not all of the TCRalphabeta and TCRgammadelta cells-expressed the CD43 S7(-) reactive determinant. CD43 S7(+) IELs secreted more IL-2, IL-4, IL-10, IL 17, and IFN-gamma following anti-CD3 stimulation, and were >4-fold more cytotoxic in fresh isolates and >16-fold more cytotoxic after anti-CD3 stimulation, than S7(-) IELs. S7(+) but not S7(-) IELs from the ileum of IL-10(-/-) mice spontaneously produced IFN-gamma. In vivo BrdU uptake by IELs in non-Ag-primed mice was greatest in the S7(+) population, indicating that significantly more S7(+) IELs than S7(-) IELs undergo cell expansion under normal homeostatic conditions. DNA microarray analyses showed that S7(+) IELs expressed higher levels of genes associated with activated T cells, whereas S7(-) IELs expressed genes used in the regulation of NK cells. These findings define two functionally distinct populations of IELs based on CD43 expression independent of TCR class, and they identify a subset of IELs that may serve as a target to better control intestinal inflammation. PMID- 15528369 TI - Genetic manipulation of telomerase in HIV-specific CD8+ T cells: enhanced antiviral functions accompany the increased proliferative potential and telomere length stabilization. AB - A large proportion of the CD8(+) T cell pool in persons chronically infected with HIV consists of cells that show features of replicative senescence, an end stage characterized by irreversible cell cycle arrest, multiple genetic and functional changes, and shortened telomeres. The objective of our research was to determine whether constitutive expression of the gene for the human telomerase (hTERT) can prevent senescence-induced impairments in human virus-specific CD8(+) T cells, particularly in the context of HIV-1 disease. Our results indicate that hTERT expressing HIV-specific CD8(+) lymphocytes show both an enhanced and sustained capacity to inhibit HIV-1 replication in in vitro coculture experiments, as well as prolonged ability to produce IFN-gamma and TNF-alpha in response to stimulation with HIV-1-derived peptides, as compared with vector-transduced controls. Loss of CD28 expression, the signature change of replicative senescence in cell culture, was retarded in those CD8(+) T cell cultures that had high levels of CD28 at the time of hTERT transduction. These findings suggest that telomere shortening may be the primary driving force behind several aspects of CD8(+) T cell dysfunction associated with replicative senescence. We also demonstrate reduced accumulation of the p16(INK4a) and p21(WAF1) cell cycle inhibitors in hTERT-transduced lymphocytes, providing a possible mechanism by which stable hTERT expression is able to circumvent the senescence barrier in CD8(+) T cells. Given the key role of CD8(+) T cell function in controlling a variety of acute and latent viral infections, approaches to retard the functional decrements associated with replicative senescence may lead to novel types of immunotherapy. PMID- 15528370 TI - Cmv1-independent antiviral role of NK cells revealed in murine cytomegalovirus infected New Zealand White mice. AB - Ly49H(+) NK cells play a critical role in innate antiviral immune responses to murine CMV (MCMV). Ly49H(b6) recognition of MCMV-encoded m157 on infected cells activates natural killing required for host resistance. We show that mAb 3D10 (anti-Ly49H) recognizes comparable subsets of NK cells from New Zealand White (NZW), New Zealand Black (NZB), and C57BL/6 spleens. However, virus levels in the spleens of MCMV-infected NZW and NZB mice differed greatly. We found that MCMV replication in infected NZW spleens was limited through NK cells. Alternately, NZB mice were profoundly susceptible to MCMV infection. Although 3D10 mAb injections given before infection interfere with Cmv1-type resistance in C57BL/6 mice, similar mAb injections did not affect NZW resistance, likely because NZW NK cell receptors did not bind MCMV-encoded m157. Instead, anti-MCMV host defenses in hybrid NZ offspring were associated with multiple chromosome locations including several putative quantitative trait loci that did not overlap with H-2 or NK gene complex loci. This study revealed a novel pathway used by NK cells to defend against MCMV infection. Thus, the importance of Ly49H in MCMV infection may be shaped by other additional background genes. PMID- 15528371 TI - Pathogen-induced apoptotic neutrophils express heat shock proteins and elicit activation of human macrophages. AB - Ingestion of aged or irradiated apoptotic neutrophils actively suppresses stimulation of macrophages (Mphi). Many bacterial pathogens can also provoke apoptosis in neutrophils, but little is known about how such apoptotic cells influence Mphi activation. We found that neutrophils undergoing apoptosis induced by UV irradiation, Escherichia coli, or Staphylococcus aureus could either stimulate or inhibit Mphi activation. In contrast to Mphi that had ingested irradiated apoptotic neutrophils, Mphi that had phagocytosed bacteria-induced apoptotic neutrophils exhibited markedly increased production of the proinflammatory cytokine TNF-alpha, but not the anti-inflammatory cytokine TGF beta. Moreover, ingestion of bacteria, but not UV-induced apoptotic neutrophils, caused increased expression of FcgammaRI on Mphi, and this effect was not provoked directly by bacteria associated with the apoptotic neutrophils. Instead, we found that a link between pathogen-induced apoptotic neutrophils and up regulation of the heat shock proteins HSP60 and HSP70, and we also observed that recombinant HSP60 and HSP70 potentiated LPS-stimulated production of TNF-alpha in Mphi. The opposing macrophage responses to neutrophils undergoing apoptosis induced in different ways may represent a novel mechanism that regulates the extent of the immune response to invading microbes in two steps: first by aiding the functions of Mphi at an early stage of infection, and subsequently by deactivating those cells through removal of uninfected apoptotic neutrophils. HSP induction in neutrophils may provide the danger signals required to generate a more effective macrophage response. PMID- 15528372 TI - Hematopoietic cells are required to initiate a Chlamydia trachomatis-specific CD8+ T cell response. AB - Chlamydia trachomatis is a global human pathogen causing diseases ranging from blinding trachoma to pelvic inflammatory disease. To explore how innate and adaptive immune responses cooperate to protect against systemic infection with C. trachomatis L2, we investigated the role of macrophages (Mphi) and dendritic cells (DCs) in the stimulation of C. trachomatis-specific CD8(+) T cells. We found that C. trachomatis infection of Mphi and DCs is far less productive than infection of nonprofessional APCs, the typical targets of infection. However, despite the limited replication of C. trachomatis within Mphi and DCs, infected Mphi and DCs process and present C. trachomatis CD8(+) T cell Ag in a proteasome dependent manner. These findings suggest that although C. trachomatis is a vacuolar pathogen, some Ags expressed in infected Mphi and DCs are processed in the host cell cytosol for presentation to CD8(+) T cells. We also show that even though C. trachomatis replicates efficiently within nonprofessional APCs both in vitro and in vivo, Ag presentation by hematopoietic cells is essential for initial stimulation of C. trachomatis-specific CD8(+) T cells. However, when DCs infected with C. trachomatis ex vivo were adoptively transferred into naive mice, they failed to prime C. trachomatis-specific CD8(+) T cells. We propose a model for priming C. trachomatis-specific CD8(+) T cells whereby DCs acquire C. trachomatis Ag by engulfing productively infected nonprofessional APCs and then present the Ag to T cells via a mechanism of cross-presentation. PMID- 15528373 TI - Multiple roles of CLAN (caspase-associated recruitment domain, leucine-rich repeat, and NAIP CIIA HET-E, and TP1-containing protein) in the mammalian innate immune response. AB - NAIP CIIA HET-E and TP1 (NACHT) family proteins are involved in sensing intracellular pathogens or pathogen-derived molecules, triggering host defense responses resulting in caspase-mediated processing of proinflammatory cytokines and NF-kappaB activation. Caspase-associated recruitment domain, leucine-rich repeat, and NACHT-containing protein (CLAN), also known as ICE protease activating factor, belongs to a branch of the NACHT family that contains proteins carrying caspase-associated recruitment domains (CARDs) and leucine-rich repeats (LRRs). By using gene transfer and RNA-interference approaches, we demonstrate in this study that CLAN modulates endogenous caspase-1 activation and subsequent IL 1beta secretion from human macrophages after exposure to LPS, peptidoglycan, and pathogenic bacteria. CLAN was also found to mediate a direct antibacterial effect within macrophages after Salmonella infection and to sensitize host cells to Salmonella-induced cell death through a caspase-1-independent mechanism. These results indicate that CLAN contributes to several biological processes central to host defense, suggesting a prominent role for this NACHT family member in innate immunity. PMID- 15528374 TI - Helminth infection protects mice from anaphylaxis via IL-10-producing B cells. AB - Modulation of the immune system by infection with helminth parasites, including schistosomes, is proposed to reduce the levels of allergic responses in infected individuals. In this study we investigated whether experimental infection with Schistosoma mansoni could alter the susceptibility of mice to an extreme allergic response, anaphylaxis. We formally demonstrate that S. mansoni infection protects mice from an experimental model of systemic fatal anaphylaxis. The worm stage of infection is shown to mediate this protective effect. In vivo depletion studies demonstrated an imperative role for B cells and IL-10 in worm-mediated protection. Furthermore, worm infection of mice increases the frequency of IL-10 producing B cells compared with that in uninfected mice. However, transfer of B cells from worm-infected mice or in vitro worm-modulated B cells to sensitized recipients exacerbated anaphylaxis, which was attributed to the presence of elevated levels of IL-4-producing B cells. Worm-modulated, IL-10-producing B cells from IL-4-deficient, but not IL-5-, IL-9- or IL-13-deficient, mice conferred complete resistance to anaphylaxis when transferred to naive mice. Therefore, we have dissected a novel immunomodulatory mechanism induced by S. mansoni worms that is dependent on an IL-10-producing B cell population that can protect against allergic hypersensitivity. These data support a role for helminth immune modulation in the hygiene hypothesis and further illustrate the delicate balance between parasite induction of protective regulatory (IL-10) responses and detrimental (IL-4) allergic responses. PMID- 15528375 TI - Single mucosal, but not parenteral, immunization with recombinant adenoviral based vaccine provides potent protection from pulmonary tuberculosis. AB - Bacillus Calmette-Guerin (BCG) vaccine has failed to control the global tuberculosis (TB) epidemic, and there is a lack of safe and effective mucosal vaccines capable of potent protection against pulmonary TB. A recombinant replication-deficient adenoviral-based vaccine expressing an immunogenic Mycobacterium tuberculosis Ag Ag85A (AdAg85A) was engineered and evaluated for its potential to be used as a respiratory mucosal TB vaccine in a murine model of pulmonary TB. A single intranasal, but not i.m., immunization with AdAg85A provided potent protection against airway Mycobacterium tuberculosis challenge at an improved level over that by cutaneous BCG vaccination. Systemic priming with an Ag85A DNA vaccine and mucosal boosting with AdAg85A conferred a further enhanced immune protection which was remarkably better than BCG vaccination. Such superior protection triggered by AdAg85 mucosal immunization was correlated with much greater retention of Ag-specific T cells, particularly CD4 T cells, in the lung and was shown to be mediated by both CD4 and CD8 T cells. Thus, adenoviral TB vaccine represents a promising novel vaccine platform capable of potent mucosal immune protection against TB. Our study also lends strong evidence that respiratory mucosal vaccination is critically advantageous over systemic routes of vaccination against TB. PMID- 15528376 TI - Apolipoprotein E modulates clearance of apoptotic bodies in vitro and in vivo, resulting in a systemic proinflammatory state in apolipoprotein E-deficient mice. AB - Apolipoprotein E (apoE) is a 34-kDa glycoprotein involved in lipoprotein transport through interaction with the low-density lipoprotein receptor and related receptors. Recently, it has become clear that apoE binding to its receptors plays a role both in development and in control of the immune system. In this study, we show that apoE modulates the rate of uptake of apoptotic cells by macrophages. In vitro, apoE-deficient macrophages ingest less apoptotic thymocytes (but not latex beads) than wild-type macrophages, and this defect can be corrected by addition of exogenous apoE protein. In vivo, the number of dying macrophages is increased in a range of tissues, including lung and brain. Possibly in response to the larger numbers of persistent apoptotic bodies, the number of live macrophages in these tissues are also increased compared with those of wild-type control mice. In addition to the significant changes in macrophage population dynamics we observed, levels of the proinflammatory cytokine TNF-alpha and the positive acute phase reactant fibrinogen are also elevated in the livers from apoE-deficient mice. In contrast, neither deletion of the gene encoding the LDL receptor nor cholesterol feeding of wild-type mice affected either the number of apoptotic bodies or the number of live macrophages. We conclude that apoE deficiency results in impaired clearance of apoptotic cell remnants and a functionally relevant systemic proinflammatory condition in mice, independent of its role in lipoprotein metabolism. Any similar reduction of apoE activity in humans may contribute to the pathogenesis of a wide range of chronic diseases including atherosclerosis, dementia, and osteoporosis. PMID- 15528377 TI - Ethanol blocks leukocyte recruitment and endothelial cell activation in vivo and in vitro. AB - Immune system impairment and increased susceptibility to infection among alcohol abusers is a significant but not well-understood problem. We hypothesized that acute ethanol administration would inhibit leukocyte recruitment and endothelial cell activation during inflammation and infection. Using LPS and carrageenan air pouch models in mice, we found that physiological concentrations of ethanol (1-5 g/kg) significantly blocked leukocyte recruitment (50-90%). Because endothelial cell activation and immune cell-endothelial cell interactions are critical regulators of leukocyte recruitment, we analyzed the effect of acute ethanol exposure on endothelial cell activation in vivo using the localized Shwartzman reaction model. In this model, ethanol markedly suppressed leukocyte accumulation and endothelial cell adhesion molecule expression in a dose-dependent manner. Finally, we examined the direct effects of ethanol on endothelial cell activation and leukocyte-endothelial cell interactions in vitro. Ethanol, at concentrations within the range found in human blood after acute exposure and below the levels that induce cytotoxicity (0.1-0.5%), did not induce endothelial cell activation, but significantly inhibited TNF-mediated endothelial cell activation, as measured by adhesion molecule (E-selectin, ICAM-1, VCAM-1) expression and chemokine (IL-8, MCP-1, RANTES) production and leukocyte adhesion in vitro. Studies exploring the potential mechanism by which ethanol suppresses endothelial cell activation revealed that ethanol blocked NF-kappaB nuclear entry in an IkappaBalpha dependent manner. These findings support the hypothesis that acute ethanol overexposure may increase the risk of infection and inhibit the host inflammatory response, in part, by blocking endothelial cell activation and subsequent immune cell-endothelial cell interactions required for efficient immune cell recruitment. PMID- 15528378 TI - Intranasal exposure of mice to house dust mite elicits allergic airway inflammation via a GM-CSF-mediated mechanism. AB - It is now well established that passive exposure to inhaled OVA leads to a state of immunological tolerance. Therefore, to elicit allergic sensitization, researchers have been compelled to devise alternative strategies, such as the systemic delivery of OVA in the context of powerful adjuvants, which are alien to the way humans are exposed and sensitized to allergens. The objectives of these studies were to investigate immune-inflammatory responses to intranasal delivery of a purified house dust mite (HDM) extract and to evaluate the role of GM-CSF in this process. HDM was delivered to BALB/c mice daily for 10 days. After the last exposure, mice were killed, bronchoalveolar lavage was performed, and samples were obtained. Expression/production of Th2-associated molecules in the lymph nodes, lung, and spleen were evaluated by real-time quantitative PCR and ELISA, respectively. Using this exposure protocol, exposure to HDM alone generated Th2 sensitization based on the expression/production of Th2 effector molecules and airway eosinophilic inflammation. Flow cytometric analysis demonstrated expansion and activation of APCs in the lung and an influx of activated Th2 effector cells. Moreover, this inflammation was accompanied by airways hyper-responsiveness and a robust memory-driven immune response. Finally, administration of anti-GM-CSF neutralizing Abs markedly reduced immune-inflammatory responses in both lung and spleen. Thus, intranasal delivery of HDM results in Th2 sensitization and airway eosinophilic inflammation that appear to be mediated, at least in part, by endogenous GM-CSF production. PMID- 15528379 TI - A novel role of cysteinyl leukotrienes to promote dendritic cell activation in the antigen-induced immune responses in the lung. AB - Although the critical role of cysteinyl leukotrienes (cysLTs) in the inflammation, especially eosinophilic lung inflammation, in asthma has been well documented, their role in the early stage of Ag-specific immune response has not been completely clarified. In the present study, with a mouse model of asthma and in vitro studies we demonstrated that cysLTs potentiated dendritic cell (DC) functions such as Ag-presenting capacity and cytokine production. The cysLT-1 receptor antagonist (LTRA) strongly suppressed the activation of these DC functions and led to inhibition of subsequent not only Th2, but also Th1, responses in the early stage of immune response. Moreover, treatment with LTRA during the early stage of the immune response potently suppressed the development of Ag inhalation-induced eosinophilic airway inflammation, mucus production, and airway hyper-reactivity in vivo. Treatment with LTRA significantly increased PGE(2) production in the lung, and treatment with the cyclooxygenase inhibitor indomethacin abolished LTRA's suppressive effect on DCs and deteriorated the Th2 and Th1 responses and airway inflammation. With in vitro studies, we also confirmed that cysLTs production by DCs increased with LPS stimulation, and that LTRA directly suppressed the alloantigen-presenting capacity of DCs. These results suggested that cysLTs potentiate DC functions both in vivo and in vitro, and that LTRA could be beneficial to suppress the initial immune response in many immune-mediated disorders beyond asthma. PMID- 15528380 TI - Platelet endothelial cell adhesion molecule deficiency or blockade significantly reduces leukocyte emigration in a majority of mouse strains. AB - PECAM is a molecule used specifically during the diapedesis step when neutrophils and monocytes leave the blood compartment. Anti-PECAM reagents, such as Abs and soluble fusion proteins, block diapedesis both in vivo and in vitro. However, the PECAM knockout mouse in C57BL/6 strain has no serious defects in most models of inflammation. We show in this study that the same PECAM knockout backcrossed into the FVB/n strain clearly has reduced leukocyte emigration in two models of inflammation. Furthermore, we show that anti-PECAM reagents can block leukocyte emigration in several other wild-type strains of mice like FVB/n, SJL, and the outbred strain Swiss Webster. This clearly shows that the C57BL/6 strain is uniquely able to compensate for the loss of PECAM function. Murine models of inflammatory disease that have been studied using C57BL/6 mice should be re evaluated using FVB/n or other mouse strains to determine whether PECAM plays a role in those models. PMID- 15528381 TI - IL-5 and granulocyte-macrophage colony-stimulating factor activate STAT3 and STAT5 and promote Pim-1 and cyclin D3 protein expression in human eosinophils. AB - Allergic inflammation is characterized by elevated eosinophil numbers and by the increased production of the cytokines IL-5 and GM-CSF, which control several eosinophil functions, including the suppression of apoptosis. The JAK/STAT pathway is important for several functions in hemopoietic cells, including the suppression of apoptosis. We report in this study that STAT3, STAT5a, and STAT5b are expressed in human eosinophils and that their signaling pathways are active following IL-5 or GM-CSF treatment. However, in airway eosinophils, the phosphorylation of STAT5 by IL-5 is reduced, an event that may be related to the reduced expression of the IL-5Ralpha on airway eosinophils. Furthermore, IL-5 and GM-CSF induced the protein expression of cyclin D3 and the kinase Pim-1, both of which are regulated by STAT-dependent processes in some cell systems. Pim-1 is more abundantly expressed in airway eosinophils than in blood eosinophils. Because Pim-1 reportedly has a role in the modulation of apoptosis, these results suggest that Pim-1 action is linked to the suppression of eosinophil apoptosis by these cytokines. Although cyclin D3 is known to be critical for cell cycle progression, eosinophils are terminally differentiated cells that do not proceed through the cell cycle. Thus, this apparent cytokine regulation of cyclin D3 suggests that there is an alternative role(s) for cyclin D3 in eosinophil biology. PMID- 15528382 TI - CD56bright NK cells are enriched at inflammatory sites and can engage with monocytes in a reciprocal program of activation. AB - Human NK cells may be divided into a CD56(dim) subset and a CD56(bright) subset. In peripheral blood, CD56(dim) NK cells dominate, whereas in lymph nodes, CD56(bright) NK cells are more common. In this study we show that CD56(bright) NK cells accumulate within inflammatory lesions in a wide variety of clinical diseases affecting several different anatomical sites. We demonstrate that when activated by the monokines IL-12, IL-15, and IL-18, these NK cells promote TNF alpha production by CD14(+) monocytes in a manner that is dependent on cell:cell contact. Conversely, CD14(+) monocytes synergize with monokines to promote IFN gamma production by these NK cells. Again, this interaction is dependent on cell:cell contact. The experiments show that CD56(bright) NK cells accumulate in inflammatory lesions and, in the appropriate cytokine environment, can engage with CD14(+) monocytes in a reciprocal activatory fashion, thereby amplifying the inflammatory response. Such a positive feedback loop is likely to be important in the pathogenesis of chronic inflammatory conditions such as rheumatoid arthritis. PMID- 15528383 TI - The class A macrophage scavenger receptor attenuates CXC chemokine production and the early infiltration of neutrophils in sterile peritonitis. AB - The macrophage scavenger receptor (SR-A) is a multifunctional receptor that is associated with several important pathological conditions, including atherosclerosis. In this study, we show, using a sterile peritonitis model, that it can regulate the inflammatory response. SR-A null mice display an increased initial granulocytic infiltration because of overproduction of the CXC chemokines, MIP-2 and keratinocyte-derived cytokine. This differential response is dependent upon particle internalization and can be mimicked by advanced glycation end product-BSA-conjugated latex beads. Thus SR-A is a nonactivating receptor, which is the first example of a pattern recognition receptor that serves to counter the activities of proinflammatory receptors and attenuates the production of specific chemokines to ensure an inflammatory response of the appropriate magnitude. PMID- 15528384 TI - Group IB secretory phospholipase A2 stimulates CXC chemokine ligand 8 production via ERK and NF-kappa B in human neutrophils. AB - Although the level of group IB secretory phospholipase A(2) (sPLA(2)-IB) has been reported to be up-regulated during inflammatory response, the role of sPLA(2)-IB on the regulation of inflammation and immune responses has not been fully elucidated. In this study, we found that sPLA(2)-IB stimulates the expression and secretion of CXCL8 without affecting other proinflammatory cytokines, such as IL 1beta or TNF alpha in human neutrophils. The induction of CXCL8 secretion by sPLA(2)-IB occurs at both the transcription and translational levels and correlates with activation of NF-kappaB. Moreover, the NF-kappaB inhibitors pyrrolidinedithiocarbamate, dexamethasone, or sulfasalazine were found to prevent CXCL8 production by sPLA(2)-IB in human neutrophils. In addition, the signaling events induced by sPLA(2)-IB included activation of the MAPK ERK and an increase in intracellular Ca(2+), which are both required for CXCL8 production. The exogenous addition of sPLA(2)-IB did not induce arachidonic acid release from human neutrophils, and the inactivation of sPLA(2)-IB by EGTA did not affect CXCL8 production by sPLA(2)-IB in human neutrophils. Taken together, we suggest that sPLA(2)-IB plays a role in the modulation of inflammatory and immune responses via the sPLA(2) receptor, by inducing CXCL8 in human neutrophils. PMID- 15528385 TI - Differential regulation of Th2 and Th1 lung inflammatory responses by protein kinase C theta. AB - In vitro and recent in vivo studies have identified protein kinase Ctheta (PKCtheta) as an important intermediate in signaling pathways leading to T cell activation, proliferation, and cytokine production. However, the importance of PKCtheta to many T cell-driven inflammatory responses has not been demonstrated. In this study we show that although PKCtheta is required for the development of a robust lung inflammatory response controlled by Th2 cells, it plays a lesser role in the development of a similar lung inflammatory response controlled by Th1 cells. PKCtheta-deficient mice were strongly compromised in generating Th2 cells and exhibited reduced airway eosinophilia and Th2 cytokine production in lungs. PKCtheta was required for the initial development of Th1 cells, with these cells exhibiting delayed kinetics of differentiation and accumulation. However, with recall Ag challenge via the airways, this defect was overcome, and lung infiltration and Th1 cytokine production were largely unimpaired in PKCtheta deficient animals. These data suggest that PKCtheta can play roles in aspects of both Th2 and Th1 responses, but lung inflammation induced by Th2 cells is more dependent on this protein kinase than lung inflammation induced by Th1 cells. PMID- 15528386 TI - Identification of selective basophil chemoattractants in human nasal polyps as insulin-like growth factor-1 and insulin-like growth factor-2. AB - In a search for novel leukocyte chemoattractants at sites of allergic inflammation, we found basophil-selective chemoattractant activity in extracts of human nasal polyps. The extracts were fractionated by reverse phase HPLC, and the resulting fractions were tested for leukocyte-stimulating activity using sensitive shape change assays. The basophil-selective activity detected was not depleted by a poxvirus CC-chemokine-binding protein affinity column. This activity was further purified by HPLC, and proteins in the bioactive fractions were analyzed by tandem electrospray mass spectrometry. Insulin-like growth factor-2 (IGF-2) was identified in these HPLC fractions, and the basophil stimulating activity was inhibited by an anti-IGF-2-neutralizing Ab. Recombinant IGF-2 induced a substantial shape change response in basophils, but not eosinophils, neutrophils, or monocytes. IGF-2 stimulated chemokinesis of basophils, but not eosinophils or neutrophils, and synergized with eotaxin 1/CCL11 in basophil chemotaxis. IGF-2 also caused up-regulation of basophil CD11b expression and inhibited apoptosis, but did not stimulate degranulation or Ca(2+) flux. Recombinant IGF-1 exhibited similar basophil-selective effects as IGF-2, and both growth factors were detected in nasal polyp extracts by ELISA. This is the first demonstration of chemokinetic factors that increase the motility of basophils, but do not act on other granulocytes or monocytes. IGF-1 and IGF-2 could play a role in the selective recruitment of basophils in vivo. PMID- 15528387 TI - Polymorphisms in the Fc epsilon RI beta promoter region affecting transcription activity: a possible promoter-dependent mechanism for association between Fc epsilon RI beta and atopy. AB - The beta subunit of the high-affinity IgE receptor (FcepsilonRI) plays an important role in IgE-mediated allergic reactions as an amplifier for cell surface expression and signal transduction of FcepsilonRI. FcepsilonRIbeta is presumed to be one of the genes linked with atopic diseases. However, the validity of the associations previously found between single nucleotide polymorphisms (SNPs) in FcepsilonRIbeta and atopic diseases is questionable. In the present study, we found correlation between the SNP of FcepsilonRIbeta at +6960A/G, resulting in a Glu237Gly amino acid substitution, and the cell surface expression level of FcepsilonRI on blood basophils, although it has been shown that the Glu237Gly mutation itself does not affect the surface expression or function of FcepsilonRI. We additionally found four SNPs in the promoter region of FcepsilonRIbeta, among which -426T/C and -654C/T were tightly linked with +6960A/G. Reporter plasmids carrying the -426C and -654T promoter displayed higher transcriptional activity than those carrying the -426T and -654C promoter. We found that transcription factor YY1 preferentially bound and transactivated the -654T promoter. Furthermore, expression of FcepsilonRI beta-chain mRNA in basophils from individuals who have the minor heterozygous genotype was significantly higher than that of the major homozygous genotype. These results suggest that the SNPs in the FcepsilonRIbeta promoter are causally linked with atopy via regulation of FcepsilonRI expression. PMID- 15528388 TI - Suppression of ongoing experimental autoimmune encephalomyelitis by neutralizing the function of the p28 subunit of IL-27. AB - IL-27 is a recently defined family member of the long-chain, four-helix bundle cytokines, which consist of EBI3, an IL-12p40-related protein, and p28, an IL 12p35-related polypeptide. The role of IL-27 in the regulation of experimental autoimmune encephalomyelitis has never been studied. We show in this study that neutralizing the in vivo function of IL-27 by Abs against IL-27 p28 rapidly suppressed an ongoing long-lasting disease in C57BL/6 mice. These Abs were then used to determine the mechanistic basis of disease suppression. We show in this study that IL-27 is involved not only in the polarization of naive T cells undergoing Ag-specific T cell activation, but also in promoting the proliferation and IFN-gamma production by polarized T cells, including the long term Th1 line that has been previously selected against the target encephalitogenic determinant. This may explain in part why neutralizing IL-27 suppresses an already established disease in a very rapid and significant manner. PMID- 15528389 TI - Immune responses of breast cancer patients to mutated epidermal growth factor receptor (EGF-RvIII, Delta EGF-R, and de2-7 EGF-R). AB - Mutated epidermal growth factor receptor (EGF-RvIII, DeltaEGF-R, and de2-7 EGF-R) is the result of an 801-bp deletion within the extracellular domain of wild-type EGF-R and is expressed by breast carcinomas, but not by normal breast tissues. EGF-RvIII is expressed both on the surface and in the cytoplasm of tumor cells. Thus, EGF-RvIII is a potential tumor-specific target for both Abs and T cells. However, it is not known whether breast cancer patients can raise immune responses to EGF-RvIII expressed by their tumors. The demonstration of EGF-RvIII specific immune responses in patients would suggest that immunization of patients with EGF-RvIII vaccines is feasible, because these vaccines may boost a pre existing immune response. We have evaluated humoral and cellular immune responses to EGF-RvIII in 16 breast cancer patients and three healthy donors. Seven of 16 patients developed EGF-RvIII-specific Abs that bound to isolated EGF-RvIII protein or the protein expressed by EGF-RvIII-transfected mouse fibroblasts. The Abs that bound to EGF-RvIII did not bind to wild-type EGF-R, and anti-EGF-RvIII Abs were not found in the sera of healthy donors. Three patients had EGF-RvIII peptide-specific lymphoproliferative responses, and two of these patients also had humoral immune responses. Humoral and cellular immune responses correlated with EGF-RvIII expression by patients' tumors in most cases. These studies demonstrate that breast cancer patients specifically recognize EGF-RvIII with an overall immune response rate of 50%, suggesting that patients may benefit from vaccination against EGF-RvIII, boosting pre-existing immune responses. PMID- 15528390 TI - Decreased expression of the Ets family transcription factor Fli-1 markedly prolongs survival and significantly reduces renal disease in MRL/lpr mice. AB - Increased Fli-1 mRNA is present in PBLs from systemic lupus erythematosus patients, and transgenic overexpression of Fli-1 in normal mice leads to a lupus like disease. We report in this study that MRL/lpr mice, an animal model of systemic lupus erythematosus, have increased splenic expression of Fli-1 protein compared with BALB/c mice. Using mice with targeted gene disruption, we examined the effect of reduced Fli-1 expression on disease development in MRL/lpr mice. Complete knockout of Fli-1 is lethal in utero. Fli-1 protein expression in heterozygous MRL/lpr (Fli-1(+/-)) mice was reduced by 50% compared with wild-type MRL/lpr (Fli-1(+/+)) mice. Fli-1(+/-) MRL/lpr mice had significantly decreased serum levels of total IgG and anti-dsDNA Abs as disease progressed. Fli-1(+/-) MRL/lpr mice had significantly increased splenic CD8(+) and naive T cells compared with Fli-1(+/+) MRL/lpr mice. Both in vivo and in vitro production of MCP-1 were significantly decreased in Fli-1(+/-) MRL/lpr mice. The Fli-1(+/-) mice had markedly decreased proteinuria and significantly lower pathologic renal scores. At 48 wk of age, survival was significantly increased in the Fli-1(+/-) MRL/lpr mice, as 100% of Fli-1(+/-) MRL/lpr mice were alive, in contrast to only 27% of Fli-1(+/+) mice. These findings indicate that Fli-1 expression is important in lupus-like disease development, and that modulation of Fli-1 expression profoundly decreases renal disease and improves survival in MRL/lpr mice. PMID- 15528391 TI - Test of the principle of initial value in rat genetic models of exercise capacity. AB - An inverse relationship between initial level of physical capacity and the magnitude of response to training is termed the principle of initial value. We tested the operation of this principle under experimental conditions of minimal genetic and environmental variation. Inbred rat strains previously identified as genetic models of low [Copenhagen (COP)] and high [Dark Agouti (DA)] intrinsic (untrained) exercise capacity were trained for 8 wk on a treadmill using two disparate protocols: 1) a relative mode where each rat exercised daily according to its initial capacity, and 2) an absolute mode where both strains received the same amount of training independent of initial capacity. Response to exercise was the change in running capacity as estimated by meters run to exhaustion before and after training. When trained with the relative mode, COP rats gained 88 m (+21%; NS) whereas DA rats increased distance run by 228 m (+36%; P < 0.001). When each strain trained with the same absolute amount of training, the COP strain showed essentially no change (-6 m, -2%) and the DA strain gained 325 m (+49%; P < 0.009). Differences in response to exercise between the COP and DA could not be explained by body mass differences, oxidative enzyme activity (citrate synthase or ATP), or spontaneous behavioral activity. Our data demonstrate that genetic factors causative of high response to exercise are not uniquely associated with genetic factors for low intrinsic capacity and thus are not in accord with the principle of initial value. PMID- 15528392 TI - Photoperiod-dependent modulation of cardiac excitation contraction coupling in the Siberian hamster. AB - In mammals, changes in photoperiod regulate a diverse array of physiological and behavioral processes, an example of which in the Siberian hamster (Phodopus sungorus) is the expression of bouts of daily torpor following prolonged exposure to a short photoperiod. During torpor, body temperature drops dramatically; however, unlike in nonhibernating or nontorpid species, the myocardium retains the ability to contract and is resistant to the development of arrhythmias. In the present study, we sought to determine whether exposure to a short photoperiod results in alterations to cardiac excitation-contraction coupling, thus potentially enabling the heart to survive periods of low temperature during torpor. Experiments were performed on single ventricular myocytes freshly isolated from the hearts of Siberian hamsters that had been exposed to either 12 wk of short-day lengths (SD) or 12 wk of long-day lengths (LD). In SD-acclimated animals, the amplitude of the systolic Ca(2+) transient was increased (e.g., from 142 +/- 17 nmol/l in LD to 229 +/- 31 nmol/l in SD at 4 Hz; P < 0.001). The increased Ca(2+) transient amplitude in the SD-acclimated animals was not associated with any change in the shape or duration of the action potential. However, sarcoplasmic reticulum Ca(2+) content measured after current-clamp stimulation was increased in the SD-acclimated animals (at 4 Hz, 110 +/- 5 vs. 141 +/- 15 mumol/l, P < 0.05). We propose that short photoperiods reprogram the function of the cardiac sarcoplasmic reticulum, resulting in an increased Ca(2+) content, and that this may be a necessary precursor for maintenance of cardiac function during winter torpor. PMID- 15528393 TI - Role of angiotensin in body fluid homeostasis of mice: effect of losartan on water and NaCl intakes. AB - It is known that mice injected peripherally with ANG II do not show a drinking response but that cFos immunoreactivity (ir) is induced in brain regions similar to those in rats. We now show in Crl:CD1(ICR) mice that peripheral injection of the ANG II type 1 receptor antagonist losartan was sufficient to prevent this induction of Fos-ir in the subfornical organ (SFO). Injection of ANG II into the lateral cerebral ventricle produced a robust water intake in mice and induced Fos ir in SFO, as well as in median preoptic (MnPO) and paraventricular (PVN) nuclei. Peripheral injection of losartan blocked this drinking response and prevented the induction of Fos-ir in each of these brain regions. Hypovolemia produced by polyethylene glycol (PEG) produced a robust water intake but no evidence of sodium appetite, and it induced Fos-ir in SFO, MnPO, and PVN. Peripheral injection of losartan did not affect this drinking response. Fos-ir induced by PEG in SFO and MnPO was reduced by treatment with losartan, while that induced in the PVN was further increased by losartan. Sodium depletion with furosemide and low-sodium diet produced a strong sodium appetite and induced Fos-ir in SFO and MnPO. Treatment with losartan completely blocked the sodium appetite, as well as the induction of Fos-ir in these brain regions. These data indicate that endogenous production of ANG II and action at forebrain receptors is critically involved in depletion-related sodium appetite in mice. The absence of an effect of losartan on PEG-induced drinking suggests the critical involvement of other factor(s) such as arterial or venous baroreceptor input, and we discuss how this factor could also explain why peripheral ANG II is not dipsogenic in mice. PMID- 15528394 TI - Intramuscular pressure-induced inhibition of cardiac contraction: implications for cardiac-locomotor synchronization. AB - The synchronization of cardiac and locomotor rhythms has been suggested to enhance the efficiency of arterial delivery to active muscles during rhythmic exercise, but direct evidence showing such a functional role has not been provided. In this study, we tested the hypothesis that the heartbeat is coupled with intramuscular pressure (IMP) changes so as to time the delivery of blood through peripheral tissues when the IMP is lower. To this end, we developed a computer-controlled, dynamic, thigh cuff occlusion device that enables bilateral thigh cuffs to repeatedly inflate and deflate, one side after the other, to simulate rhythmic IMP changes during bipedal locomotion. Nine healthy subjects were examined, and three different occlusion pressures (50, 80, and 120 mmHg) were applied separately to the thigh cuffs of normal subjects while they were sitting. Alternate occlusions of the bilateral thigh cuffs administered at the frequency of the mean heart rate produced significant phase synchronization between the cardiac and cuff-occlusion rhythms when 120 mmHg pressure was applied. However, synchronization was not observed when the occlusion pressure was 50 or 80 mmHg. During synchronization, heartbeats were most likely to occur in phases that did not include overlap between the peak arterial flow velocity in the thigh and elevated cuff pressure. We believe that phase synchronization occurs so that the cardiac cycle is timed to deliver blood through the lower legs when IMP is not maximal. If this can be extrapolated to natural locomotion, synchronization between cardiac and locomotor activities may be associated with the improved perfusion of exercising muscles. PMID- 15528395 TI - Cortisol-induced CXCR4 augmentation mobilizes T lymphocytes after acute physical stress. AB - The aim of this study was to elucidate the mechanism responsible for lymphopenia after exercise. Seven young healthy men volunteered for this study. Peripheral blood mononuclear cells (PBMC) were cultured with cortisol and analyzed for C-X-C motif chemokine receptor 4 (CXCR4) expression by flow cytometry. To determine the effects of exercise, subjects performed exhaustive cycling exercise. PBMC were cultured with plasma obtained before and after the cycling exercise. Alternatively, PBMC obtained before and after exercise were cultured without plasma or glucocorticoid to examine whether PBMC were primed in vivo for CXCR4 expression. We analyzed cortisol- or plasma-treated PBMC to determine their ability to migrate through membrane filters in response to stromal cell-derived factor 1alpha/CXCL12. Cortisol dose- and time-dependently augmented CXCR4 expression on T lymphocytes, with <6 h of treatment sufficient to augment CXCR4 on T lymphocytes. Postexercise plasma also augmented CXCR4 expression. Cortisol or postexercise plasma treatment markedly enhanced migration of T lymphocytes toward CXCL12. Augmentation of CXCR4 on T lymphocytes by cortisol or plasma was effectively blocked by the glucocorticoid receptor antagonist RU-486. Thus exercise-elicited endogenous cortisol effectively augments CXCR4 expression on T lymphocytes, which may account for lymphopenia after exercise. PMID- 15528396 TI - Differential effects of maternal hypoxia or nutrient restriction on carotid and femoral vascular function in neonatal rats. AB - In response to reduced oxygen or nutrient supply, the fetus may redistribute cardiac output to conserve brain and heart growth, at the expense of the peripheral tissues; however, it is not known whether alterations in vascular function are maintained after birth or whether reduced fetal oxygen versus nutrient supply produces distinct effects. Using a pressure myograph, we examined isolated carotid and femoral artery responses to phenylephrine and endothelin-1 in neonatal rats, after either reduced maternal oxygen or global nutrient restriction during late gestation. Timed-pregnant Sprague-Dawley rats were randomly assigned to control (n = 10), hypoxia (12% O2, n = 9), or nutrient restriction (NR, 40% of control diet, n = 7) protocol and treated from day 15-21 of pregnancy. Pups were collected 3-12 h after birth. Neonatal weights (P < 0.001) and relative liver weights (P < 0.001) were lower in hypoxia and nutrient restriction treatments compared with control, while relative heart weights were greater in the hypoxia than in the control or nutrient restriction groups (P < 0.01). Constriction to phenylephrine was reduced in carotid arteries from the hypoxia and nutrient restriction groups compared with control (P < 0.001), while the femoral artery response was greater in hypoxia-treated neonates compared with control or nutrient-restricted neonates (P < 0.01). Only the hypoxia reduced carotid responses to endothelin-1, while no differences were observed in the endothelin-1 responses in femoral arteries. Maternal hypoxia and maternal nutrient restriction produced distinct effects on heart growth and neonatal vascular function, suggesting that regional changes in cardiovascular function after poor fetal growth are dependent on the nature of the insult in utero. PMID- 15528397 TI - Heme oxygenase-derived carbon monoxide promotes arteriolar endothelial dysfunction and contributes to salt-induced hypertension in Dahl salt-sensitive rats. AB - Vascular tissues express heme oxygenase (HO), which metabolizes heme to form carbon monoxide (CO). Heme-derived CO inhibits nitric oxide synthase and promotes endothelium-dependent vasoconstriction. After 4 wk of high-salt diet, Dahl salt sensitive (Dahl-S) rats display hypertension, increased vascular HO-1 expression, and attenuated vasodilator responses to ACh that can be completely restored by acute treatment with an inhibitor of HO. In this study, we examined the temporal development of HO-mediated endothelial dysfunction in isolated pressurized first order gracilis muscle arterioles, identified the HO product responsible, and studied the blood pressure effects of HO inhibition in Dahl-S rats on a high-salt diet. Male Dahl-S rats (5-6 wk) were placed on high-salt (8% NaCl) or low-salt (0.3% NaCl) diets for 0-4 wk. Blood pressure increased gradually, and responses to an endothelium-dependent vasodilator, ACh, decreased gradually with the length of high-salt diet. Flow-induced dilation was abolished in hypertensive Dahl-S rats. Acute in vitro pretreatment with an inhibitor of HO, chromium mesoporphyrin (CrMP), restored endothelium-dependent vasodilation and abolished the differences between groups. The HO product CO prevented the restoration of endothelium dependent dilation by CrMP. Furthermore, administration of an HO inhibitor lowered blood pressure in Dahl-S rats with salt-induced hypertension but did not do so in low-salt control rats. These results suggest that hypertension and HO mediated endothelial dysfunction develop gradually and simultaneously in Dahl-S rats on high-salt diets. They also suggest that HO-derived CO underlies the impaired endothelial dysfunction and contributes to hypertension in Dahl-S rats on high-salt diets. PMID- 15528398 TI - Neuroendocrinology of nutritional infertility. AB - Natural selection has linked the physiological controls of energy balance and fertility such that reproduction is deferred during lean times, particularly in female mammals. In this way, an energetically costly process is confined to periods when sufficient food is available to support pregnancy and lactation. Even in the face of abundance, nutritional infertility ensues if energy intake fails to keep pace with expenditure. A working hypothesis is proposed in which any activity or condition that limits the availability of oxidizable fuels (e.g., undereating, excessive energy expenditure, diabetes mellitus) can inhibit both gonadotropin-releasing hormone (GnRH)/luteinizing hormone secretion and female copulatory behaviors. Decreases in metabolic fuel availability appear to be detected by cells in the caudal hindbrain. Hindbrain neurons producing neuropeptide Y (NPY) and catecholamines (CA) then project to the forebrain where they contact GnRH neurons both directly and also indirectly via corticotropin releasing hormone (CRH) neurons to inhibit GnRH secretion. In the case of estrous behavior, the best available evidence suggests that the inhibitory NPY/CA system acts primarily via CRH or urocortin projections to various forebrain loci that control sexual receptivity. Disruption of these signaling processes allows normal reproduction to proceed in the face of energetic deficits, indicating that the circuitry responds to energy deficits and that no signal is necessary to indicate that there is an adequate energy supply. While there is a large body of evidence to support this hypothesis, the data do not exclude nutritional inhibition of reproduction by other pathways and processes, and the full story will undoubtedly be more complex than this. PMID- 15528399 TI - The drive to regain is mainly in the brain. PMID- 15528400 TI - Tonic glutamatergic drive of RVLM vasomotor neurons? PMID- 15528401 TI - Aerobic performance and the function of myoglobin in human skeletal muscle. PMID- 15528402 TI - Metabolic and vascular support for the role of myoglobin in humans: a multiparametric NMR study. AB - In human muscle the role of myoglobin (Mb) and its relationship to factors such as muscle perfusion and metabolic capacity are not well understood. We utilized nuclear magnetic resonance (NMR) to simultaneously study the Mb concentration ([Mb]), perfusion, and metabolic characteristics in calf muscles of athletes trained long term for either sprint or endurance running after plantar flexion exercise and cuff ischemia. The acquisitions for (1)H assessment of Mb desaturation and concentration, arterial spin labeling measurement of muscle perfusion, and (31)P spectroscopy to monitor high-energy phosphate metabolites were interleaved in a 4-T magnet. The endurance-trained runners had a significantly elevated [Mb] (0.28 +/- 0.06 vs. 0.20 +/- 0.03 mmol/kg). The time constant of creatine rephosphorylation (tauPCr), an indicator of oxidative capacity, was both shorter in the endurance-trained group (34 +/- 6 vs. 64 +/- 20 s) and negatively correlated with [Mb] across all subjects (r = 0.58). The time to reach maximal perfusion after cuff release was also both shorter in the endurance-trained group (306 +/- 74 vs. 560 +/- 240 s) and negatively correlated with [Mb] (r = 0.56). Finally, Mb reoxygenation rate tended to be higher in the endurance-trained group and was positively correlated with tauPCr (r = 0.75). In summary, these NMR data reveal that [Mb] is increased in human muscle with a high oxidative capacity and a highly responsive vasculature, and the rate at which Mb resaturates is well correlated with the rephosphorylation rate of Cr, each of which support a teleological role for Mb in O(2) transport within highly oxidative human skeletal muscle. PMID- 15528403 TI - E-ring 8-isoprostanes are agonists at EP2- and EP4-prostanoid receptors on human airway smooth muscle cells and regulate the release of colony-stimulating factors by activating cAMP-dependent protein kinase. AB - 8-Isoprostanes are bioactive lipid mediators formed via the nonenzymatic peroxidation of arachidonic acid by free radicals and reactive oxygen species. However, their cognate receptors, biological actions, and signaling pathways are poorly studied. Here, we report the effect of a variety of E- and Falpha-ring 8 isoprostanes on the release of granulocyte/macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) from human airway smooth muscle (HASM) cells stimulated with interleukin-1beta (IL-1beta). The elaboration of GM-CSF and G-CSF by IL-1beta was inhibited and augmented, respectively, in a concentration-dependent manner by 8-iso-prostaglandin (PG) E(1) and 8-iso-PGE(2), but not by 8-iso-PGF(1alpha), 8-iso-PGF(2alpha), and 8-iso PGF(3)alpha. AH 6809 (6-isopropoxy-9-oxoxanthine-2-carboxylic acid), an EP(1) /EP(2)-/DP-receptor blocking drug, antagonized the inhibitory effect of 8-iso PGE(1) and 8-iso-PGE(2) on GM-CSF output with an affinity consistent with an interaction at prostanoid receptors of the EP(2)-subtype. In contrast, the facilitation by 8-iso-PGE(1) and 8-iso-PGE(2) of G-CSF release was unaffected by AH 6809 and the selective EP(4)-receptor antagonist L-161,982 [4'-[3-butyl-5-oxo 1-(2-trifluoromethyl-phenyl)-1,5-dihydro-[1,2,4]triazol-4-ylmethyl]-biphenyl-2 sulfonic acid (3-methyl-thiophene-2-carbonyl)-amide]. However, when used in combination, AH 6809 and L-161,982 displaced 5-fold to the right the 8-iso-PGE and 8-iso-PGE concentration-response curves. The opposing (1)effect of E-ring (2)8-isoprostanes on GM-CSF and G-CSF release was mimicked by 8-bromo-cAMP and abolished in cells infected with an adenovirus vector encoding an inhibitor protein of cAMP-dependent protein kinase (PKA). Together, these data demonstrate that E-ring 8-isoprostanes regulate the secretion of GM-CSF and G-CSF from HASM cells by a cAMP- and PKA-dependent mechanism. Moreover, antagonist studies revealed that 8-iso-PGE(1) and 8-iso-PGE(2) act solely via EP(2) -receptors to inhibit GM-CSF release, whereas both EP(2)- and EP(4)-receptor subtypes positively regulate G-CSF output. PMID- 15528404 TI - Examination of human tissue cytosols for expression of sulfotransferase isoform 1A2 (SULT1A2) using a SULT1A2-specific antibody. AB - Sulfotransferase isoform 1A2 (SULT1A2) is a member of the cytosolic sulfotransferase family of phase II detoxification enzymes. Studies with recombinant enzymes have shown that SULT1A2 can catalyze the bioactivation of several procarcinogens, indicating a potential role in chemical carcinogenesis. However, previous studies have suggested that the SULT1A2 transcript has a splicing defect that might prevent it from becoming translated into protein; therefore, we sought to determine the expression of SULT1A2 in tissues. An antibody directed against a region of human SULT1A2 that differs from other known sulfotransferase isoforms was developed and used to screen a large number of cytosolic fractions from various tissues. Although the SULT1A2 antibody recognized recombinant SULT1A2 and did not cross-react with other SULT isoforms, the expression of SULT1A2 was not detected in any tissue examined. These studies suggest that if SULT1A2 is expressed as protein, the levels are very low and that SULT1A2 probably does not play a physiological role in chemical carcinogenesis. PMID- 15528405 TI - Y chromosome of D. pseudoobscura is not homologous to the ancestral Drosophila Y. AB - We report a genome-wide search of Y-linked genes in Drosophila pseudoobscura. All six identifiable orthologs of the D. melanogaster Y-linked genes have autosomal inheritance in D. pseudoobscura. Four orthologs were investigated in detail and proved to be Y-linked in D. guanche and D. bifasciata, which shows that less than 18 million years ago the ancestral Drosophila Y chromosome was translocated to an autosome in the D. pseudoobscura lineage. We found 15 genes and pseudogenes in the current Y of D. pseudoobscura, and none are shared with the D. melanogaster Y. Hence, the Y chromosome in the D. pseudoobscura lineage appears to have arisen de novo and is not homologous to the D. melanogaster Y. PMID- 15528406 TI - Hemoxygenase-2 is an oxygen sensor for a calcium-sensitive potassium channel. AB - Modulation of calcium-sensitive potassium (BK) channels by oxygen is important in several mammalian tissues, and in the carotid body it is crucial to respiratory control. However, the identity of the oxygen sensor remains unknown. We demonstrate that hemoxygenase-2 (HO-2) is part of the BK channel complex and enhances channel activity in normoxia. Knockdown of HO-2 expression reduced channel activity, and carbon monoxide, a product of HO-2 activity, rescued this loss of function. Inhibition of BK channels by hypoxia was dependent on HO-2 expression and was augmented by HO-2 stimulation. Furthermore, carotid body cells demonstrated HO-2-dependent hypoxic BK channel inhibition, which indicates that HO-2 is an oxygen sensor that controls channel activity during oxygen deprivation. PMID- 15528407 TI - Rescue of dystrophic muscle through U7 snRNA-mediated exon skipping. AB - Most mutations in the dystrophin gene create a frameshift or a stop in the mRNA and are associated with severe Duchenne muscular dystrophy. Exon skipping that naturally occurs at low frequency sometimes eliminates the mutation and leads to the production of a rescued protein. We have achieved persistent exon skipping that removes the mutated exon on the dystrophin messenger mRNA of the mdx mouse, by a single administration of an AAV vector expressing antisense sequences linked to a modified U7 small nuclear RNA. We report the sustained production of functional dystrophin at physiological levels in entire groups of muscles and the correction of the muscular dystrophy. PMID- 15528408 TI - Acetylation by Tip60 is required for selective histone variant exchange at DNA lesions. AB - Phosphorylation of the human histone variant H2A.X and H2Av, its homolog in Drosophila melanogaster, occurs rapidly at sites of DNA double-strand breaks. Little is known about the function of this phosphorylation or its removal during DNA repair. Here, we demonstrate that the Drosophila Tip60 (dTip60) chromatin remodeling complex acetylates nucleosomal phospho-H2Av and exchanges it with an unmodified H2Av. Both the histone acetyltransferase dTip60 as well as the adenosine triphosphatase Domino/p400 catalyze the exchange of phospho-H2Av. Thus, these data reveal a previously unknown mechanism for selective histone exchange that uses the concerted action of two distinct chromatin-remodeling enzymes within the same multiprotein complex. PMID- 15528409 TI - By carrot or by stick: cognitive reinforcement learning in parkinsonism. AB - To what extent do we learn from the positive versus negative outcomes of our decisions? The neuromodulator dopamine plays a key role in these reinforcement learning processes. Patients with Parkinson's disease, who have depleted dopamine in the basal ganglia, are impaired in tasks that require learning from trial and error. Here, we show, using two cognitive procedural learning tasks, that Parkinson's patients off medication are better at learning to avoid choices that lead to negative outcomes than they are at learning from positive outcomes. Dopamine medication reverses this bias, making patients more sensitive to positive than negative outcomes. This pattern was predicted by our biologically based computational model of basal ganglia-dopamine interactions in cognition, which has separate pathways for "Go" and "NoGo" responses that are differentially modulated by positive and negative reinforcement. PMID- 15528410 TI - Decisive day for European research. PMID- 15528411 TI - Planetary science. Titan remains mysterious with a hint of the familiar. PMID- 15528412 TI - Infectious diseases. Ducks may magnify threat of avian flu virus. PMID- 15528413 TI - Science policy. Nanotech forum aims to head off replay of past blunders. PMID- 15528414 TI - FRENCH protests. Researchers back a 70-page agenda to reform agencies, boost careers. PMID- 15528415 TI - High-energy astrophysics. Hot on the trail of cosmic rays. PMID- 15528416 TI - Russian Academy of Sciences. Putin: reform begins at home. PMID- 15528418 TI - Science policy. Japan to host global forum on societal impacts. PMID- 15528417 TI - European Union. New research commissioner caught in controversy's wake. PMID- 15528419 TI - Plant biology. NO-making enzyme no more: Cell, PNAS papers retracted. PMID- 15528420 TI - Infectious diseases. Ethicists to guide rationing of flu vaccine. PMID- 15528421 TI - Toxicology. EPA criticized for study of child pesticide exposure. PMID- 15528422 TI - Paleontology. Dinosaurs under the knife. PMID- 15528423 TI - Cancer research. Inflammation and cancer: the link grows stronger. PMID- 15528424 TI - Invasive species. Expanding trade with China creates ecological backlash. PMID- 15528425 TI - Remediating Rocky Flats. PMID- 15528426 TI - Net environmental benefit analysis. PMID- 15528427 TI - Support for Steiger's policies. PMID- 15528428 TI - The Gulf of Mexico's dead zone. PMID- 15528429 TI - Comment on "The origins of genome complexity". PMID- 15528431 TI - Neuroscience. What genes tell us about nicotine addiction. PMID- 15528432 TI - Physics. Molecular plasmonics. PMID- 15528433 TI - Physics. The best materials for tiny, clever sensors. PMID- 15528434 TI - Microbiology. Dynamic instability of a bacterial engine. PMID- 15528435 TI - Autophagy in health and disease: a double-edged sword. AB - Autophagy, the process by which cells recycle cytoplasm and dispose of excess or defective organelles, has entered the research spotlight largely owing to the discovery of the protein components that drive this process. Identifying the autophagy genes in yeast and finding orthologs in other organisms reveals the conservation of the mechanism of autophagy in eukaryotes and allows the use of molecular genetics and biology in different model systems to study this process. By mostly morphological studies, autophagy has been linked to disease processes. Whether autophagy protects from or causes disease is unclear. Here, we summarize current knowledge about the role of autophagy in disease and health. PMID- 15528436 TI - Uridine addition after microRNA-directed cleavage. AB - One of the important roles of microRNA (miRNA) is to direct the cleavage of messenger RNA (mRNA). However, the mechanisms of decay of the cleaved mRNA products is not well understood. We show that miRNA-directed cleavage products in organisms as diverse as Arabidopsis, mouse, and Epstein-Barr virus have at their 3' ends a stretch (1 to 24 nucleotides) of oligouridine posttranscriptionally added downstream of the cleavage site. This 3' uridine addition, as shown for Arabidopsis, is correlated with decapping and 5' shortening of the cleaved products, suggesting a mechanistic step in the miRNA-directed mRNA decay mechanism. PMID- 15528437 TI - Jet and accretion-disk emission untangled in 3C 273. AB - A long-term spectral variability study of 3C 273 on data obtained by BeppoSAX allows us to decouple the beamed nonthermal (jet) and unbeamed thermal (accretion flow) radiation produced in the inner region of a radio-loud active galactic nucleus. Jet power, when compared with unbeamed radiation, increases with energy. The thermal component is generally overwhelmed by the nonthermal radiation, by a factor of 1.2 to 3 in the 2- to 10-keV range and up to a factor of 7 above 20 keV. In only one case, the accretion flow overcomes the jet, allowing the K(alpha) iron line to emerge clearly over the continuum. PMID- 15528438 TI - Energy transfer across a metal film mediated by surface plasmon polaritons. AB - Coupled surface plasmon polaritons (SPPs) are shown to provide effective transfer of excitation energy from donor molecules to acceptor molecules on opposite sides of metal films up to 120 nanometers thick. This variant of radiative transfer should allow directional control over the flow of excitation energy with the use of suitably designed metallic nanostructures, with SPPs mediating transfer over length scales of 10(-7) to 10(-4) meters. In the emerging field of nanophotonics, such a prospect could allow subwavelength-scale manipulation of light and provide an interface to the outside world. PMID- 15528439 TI - Enhancement of ferroelectricity in strained BaTiO3 thin films. AB - Biaxial compressive strain has been used to markedly enhance the ferroelectric properties of BaTiO3 thin films. This strain, imposed by coherent epitaxy, can result in a ferroelectric transition temperature nearly 500 degrees C higher and a remanent polarization at least 250% higher than bulk BaTiO3 single crystals. This work demonstrates a route to a lead-free ferroelectric for nonvolatile memories and electro-optic devices. PMID- 15528440 TI - Cation exchange reactions in ionic nanocrystals. AB - Cation exchange has been investigated in a wide range of nanocrystals of varying composition, size, and shape. Complete and fully reversible exchange occurs, and the rates of the reactions are much faster than in bulk cation exchange processes. A critical size has been identified below which the shapes of complex nanocrystals evolve toward the equilibrium shape with lowest energy during the exchange reaction. Above the critical size, the anion sublattice remains intact and the basic shapes of the initial nanocrystals are retained throughout the cation exchange. The size-dependent shape change can also be used to infer features of the microscopic mechanism. PMID- 15528441 TI - Biodiversity effects on soil processes explained by interspecific functional dissimilarity. AB - The loss of biodiversity can have significant impacts on ecosystem functioning, but the mechanisms involved lack empirical confirmation. Using soil microcosms, we show experimentally that functional dissimilarity among detritivorous species, not species number, drives community compositional effects on leaf litter mass loss and soil respiration, two key soil ecosystem processes. These experiments confirm theoretical predictions that biodiversity effects on ecosystem functioning can be predicted by the degree of functional differences among species. PMID- 15528442 TI - Dynamic instability in a DNA-segregating prokaryotic actin homolog. AB - Dynamic instability-the switching of a two-state polymer between phases of steady elongation and rapid shortening-is essential to the cellular function of eukaryotic microtubules, especially during chromosome segregation. Since the discovery of dynamic instability 20 years ago, no other biological polymer has been found to exhibit this behavior. Using total internal reflection fluorescence microscopy and fluorescence resonance energy transfer, we observe that the prokaryotic actin homolog ParM, whose assembly is required for the segregation of large, low-copy number plasmids, displays both dynamic instability and symmetrical, bidirectional polymerization. The dynamic instability of ParM is regulated by adenosine triphosphate (ATP) hydrolysis, and filaments are stabilized by a cap of ATP-bound monomers. ParM is not related to tubulin, so its dynamic instability must have arisen by convergent evolution driven by a set of common constraints on polymer-based segregation of DNA. PMID- 15528443 TI - Nicotine activation of alpha4* receptors: sufficient for reward, tolerance, and sensitization. AB - The identity of nicotinic receptor subtypes sufficient to elicit both the acute and chronic effects of nicotine dependence is unknown. We engineered mutant mice with a4 nicotinic subunits containing a single point mutation, Leu9' --> Ala9' in the pore-forming M2 domain, rendering a4* receptors hypersensitive to nicotine. Selective activation of a4* nicotinic acetylcholine receptors with low doses of agonist recapitulates nicotine effects thought to be important in dependence, including reinforcement in response to acute nicotine administration, as well as tolerance and sensitization elicited by chronic nicotine administration. These data indicate that activation of a4* receptors is sufficient for nicotine-induced reward, tolerance, and sensitization. PMID- 15528444 TI - MHC class I peptides as chemosensory signals in the vomeronasal organ. AB - The mammalian vomeronasal organ detects social information about gender, status, and individuality. The molecular cues carrying this information remain largely unknown. Here, we show that small peptides that serve as ligands for major histocompatibility complex (MHC) class I molecules function also as sensory stimuli for a subset of vomeronasal sensory neurons located in the basal Gao- and V2R receptor-expressing zone of the vomeronasal epithelium. In behaving mice, the same peptides function as individuality signals underlying mate recognition in the context of pregnancy block. MHC peptides constitute a previously unknown family of chemosensory stimuli by which MHC genotypic diversity can influence social behavior. PMID- 15528445 TI - Autophagy defends cells against invading group A Streptococcus. AB - We found that the autophagic machinery could effectively eliminate pathogenic group A Streptococcus (GAS) within nonphagocytic cells. After escaping from endosomes into the cytoplasm, GAS became enveloped by autophagosome-like compartments and were killed upon fusion of these compartments with lysosomes. In autophagy-deficient Atg5-/- cells, GAS survived, multiplied, and were released from the cells. Thus, the autophagic machinery can act as an innate defense system against invading pathogens. PMID- 15528446 TI - Structural insights into the assembly of the type III secretion needle complex. AB - Type III secretion systems (TTSSs) mediate translocation of virulence factors into host cells. We report the 17-angstrom resolution structures of a central component of Salmonella typhimurium TTSS, the needle complex, and its assembly precursor, the bacterial envelope-anchored base. Both the base and the fully assembled needle complex adopted multiple oligomeric states in vivo, and needle assembly was accompanied by recruitment of the protein PrgJ as a structural component of the base. Moreover, conformational changes during needle assembly created scaffolds for anchoring both PrgJ and the needle substructure and may provide the basis for substrate-specificity switching during type III secretion. PMID- 15528447 TI - Tracking SNARE complex formation in live endocrine cells. AB - Syntaxin, synaptosome-associated protein of 25 kD (SNAP25), and vesicle associated membrane protein/synaptobrevin are collectively called SNAP receptor (SNARE) proteins, and they catalyze neuronal exocytosis by forming a "core complex." The steps in core complex formation are unknown. Here, we monitored SNARE complex formation in vivo with the use of a fluorescent version of SNAP25. In PC12 cells, we found evidence for a syntaxin-SNAP25 complex that formed with high affinity, required only the amino-terminal SNARE motif of SNAP25, tolerated a mutation that blocks formation of other syntaxin-SNAP25 complexes, and assembled reversibly when Ca2+ entered cells during depolarization. The complex may represent a precursor to the core complex formed during a Ca2+-dependent priming step of exocytosis. PMID- 15528448 TI - A link between mRNA turnover and RNA interference in Arabidopsis. AB - In RNA interference (RNAi), double-stranded RNA (dsRNA) triggers degradation of homologous messenger RNA. In many organisms, RNA-dependent RNA polymerase (RdRp) is required to initiate or amplify RNAi, but the substrate for dsRNA synthesis in vivo is not known. Here, we show that RdRp-dependent transgene silencing in Arabidopsis was caused by mutation of XRN4, which is a ribonuclease (RNase) implicated in mRNA turnover by means of decapping and 5'-3' exonucleolysis. When both XRN4 and the RdRp were mutated, the plants accumulated decapped transgene mRNA. We propose that mRNAs lacking a cap structure become exposed to RdRp to initiate or maintain RNAi. PMID- 15528449 TI - Prostaglandin D2-induced eosinophilic airway inflammation is mediated by CRTH2 receptor. AB - Mast cell-derived prostaglandin D(2) (PGD(2)) is one of the essential modulators of eosinophilic airway inflammation in asthma and allergic rhinitis. Two G protein-coupled receptors for PGD(2), prostaglandin D(2) receptor (DP) and chemoattractant receptor-homologous molecule expressed on Th(2) cells (CRTH2), are both expressed on the surface of eosinophils, and CRTH2 has been demonstrated to mediate PGD(2)-induced eosinophil mobilization in vitro. However, it has not yet been determined whether PGD(2) and its receptors mediate in vivo eosinophil trafficking into the airways or other organs. We demonstrated that intratracheal administration of PGD(2) in rats pretreated with systemic interleukin-5 (IL-5) injection induced marked airway eosinophilia, determined by the differential counts of cells in bronchoalveolar lavage (BAL) fluid and lung histology, within 2 h. Systemic IL-5 alone significantly increased the number of eosinophils in the peripheral blood but showed no effect on airway eosinophilia. Three CRTH2 specific agonists (13,14-dihydro-15-keto-PGD(2), 11-deoxy-11-methylene-15-keto PGD(2), and indomethacin) demonstrated equivalent induction of BAL eosinophilia to that of PGD(2), but a DP agonist (BW 245C [5-(6-carboxyhexyl)-1-(3-cyclohexyl 3-hydroxypropyl)-hydantoin]) or a thromboxane A(2) receptor (TP) agonist ([1S 1alpha,2beta(5Z), 3alpha(1E,3R*),4alpha)]-7-[3-(3-hydroxy-4-(4'-iodophenoxy)-1 butenyl)-7-oxabicyclo-[2.2.1]heptan-2-yl]-5-heptenoic acid) showed no effect. PGD(2) or CRTH2 agonist-induced BAL eosinophilia was almost completely inhibited by pretreatment with a CRTH2/TP antagonist, ramatroban [BAY-u3405; (+)-(3R)-3-(4 fluorobenzenesulfonamido)-1,2,3,4-tetra-hydrocarbazole-9-propionic acid], whereas a TP-specific antagonist, SQ29,548 (5-heptenoic, 7-[3-[[2 [(phenylamino)carbonyl]hydrazino]methyl]-7-oxabicyclo[2.2.1]-hept-2-yl]-[1S [1alpha,2alpha(Z),3alpha,4alpha]]), or a DP-specific antagonist, BW A868C [3 benzyl-5-(6-carboxyhexyl)-1-(2-cyclohexy-2-hydroxyethylamino)-hydantoin], did not inhibit the effects of PGD(2). These results suggest that CRTH2 plays a significant role in the eosinophil trafficking from the bloodstream into the airways in PGD(2)-related airway inflammation. PMID- 15528450 TI - Dual, hyperalgesic, and analgesic effects of the high-efficacy 5 hydroxytryptamine 1A (5-HT1A) agonist F 13640 [(3-chloro-4-fluoro-phenyl)-[4 fluoro-4-{[(5-methyl-pyridin-2-ylmethyl)-amino]-methyl}piperidin-1-yl]methanone, fumaric acid salt]: relationship with 5-HT1A receptor occupancy and kinetic parameters. AB - The aim of the present study was to establish the relationship between the plasma and brain concentration-time profiles of F 13640 [(3-chloro-4-fluoro-phenyl)-[4 fluoro-4-{[(5-methyl-pyridin-2-ylmethyl)-amino]-methyl}piperidin-1-yl]methanone, fumaric acid salt] after acute administration and both its hyper- and hypoanalgesic effects in rats. The maximal plasma concentration (C(max)) of F 13640 after i.p. administration of 0.63 mg/kg was obtained at 15 min and decreased to half its maximal value after about 1 h. The amount of F 13640 collected by means of in vivo microdialysis in hippocampal dialysates could be measured reliably after 0.63 and 2.5 mg/kg, reached its maximum at about 1 h, and fell to half of its maximal value at about 3 h. 5-Hydroxytryptamine 1A (5-HT(1A)) receptor occupancy was estimated by ex vivo binding in rat brain sections. F 13640 inhibited [(3)H]8-hydroxy-2-[di-n-propylamino] tetralin binding ex vivo in rat hippocampus, entorhinal cortex, and frontal cortex (ED(50), 0.34 mg/kg i.p.). Maximal inhibition was reached at approximately 30 min after 0.63 mg/kg F 13640 and fell to half of its value after about 4 to 8 h. After injection (15 min) in the paw pressure test, F 13640 (0.63 mg/kg i.p.) induced an initial hyperalgesia that was followed 4 h later by a paradoxical analgesia that lasted until 8 h. In contrast, in the formalin test, F 13640 inhibited pain behaviors until 4 h after drug administration. F 13640 also produced elements of the 5-HT syndrome that lasted up to 4 h after administration. These results demonstrate that F 13640 induces hyperalgesia and/or analgesia with a time course that parallels the occupancy of 5-HT(1A) receptors and the presence of the compound in blood and brain. PMID- 15528451 TI - P-glycoprotein and mutlidrug resistance-associated proteins limit the brain uptake of saquinavir in mice. AB - Efflux transporters such as P-glycoprotein (P-gp) and multidrug resistance associated proteins (Mrps) and their contributions to saquinavir (SQV) brain uptake were characterized. Cerebral flow rate was estimated from diazepam uptake and brain vascular volume was assessed using inulin. Mice brains were perfused with buffer containing SQV alone or coperfused with different concentrations of GF120918, a P-gp inhibitor or MK571, a specific Mrp family inhibitor. Inulin, a nonabsorbable marker, was also coperfused in all studies to assess whether the inhibitors altered the physical integrity of the blood-brain barrier (BBB). The estimated cerebral flow rate using diazepam was 250 ml.100g(-1.)min(-1). The brain vascular volume, estimated using inulin, was almost constant (0.94 +/- 0.03 ml.100 g(-1), n = 12) during the perfusion study. SQV uptake kinetics was linear during the sampling period. Inclusion of 10 muM GF120918 in the perfusate resulted in a more than 7-fold increase in the brain distributional volume (i.e., uptake) of SQV. Inclusion of 100 muM MK571 in the perfusate increased SQV apparent brain uptake by more than 4.4-fold, suggesting, for the first time, that Mrp transporters may play an important role in the brain uptake and retention of SQV. Neither GF120918 nor MK571 altered the integrity of the BBB during the time course of the study. Although the current results reaffirm that SQV is a P-gp substrate, this is the first report implicating the Mrp transporter family in the limited brain uptake and retention of SQV in vivo in mice. PMID- 15528452 TI - In vitro metabolism of nitric oxide-donating aspirin: the effect of positional isomerism. AB - NO-donating aspirin (NO-ASA) is a potentially important chemopreventive agent against cancer. Since positional isomerism affects strongly its potency in inhibiting colon cancer cell growth, we studied the metabolic transformations of its ortho-, meta-, and para-isomers in rat liver and colon cytosolic, microsomal, and mitochondrial fractions as well as in intact HT-29 human colon cancer cells. NO-ASA and metabolites were determined by high-performance liquid chromatography and products identified by mass spectroscopy, as required. For all three isomers, the acetyl group on the ASA moiety was hydrolyzed rapidly. This was followed by hydrolysis of the ester bond linking the salicylate anion to the spacer. The ortho- and para-isomers produced salicylic acid and a putative intermediate consisting of the remainder of the molecule, which via a rapid step generated nitrate, (hydroxymethyl)phenol, and a conjugate of spacer with glutathione. The meta-isomer, in contrast, generated salicylic acid and (nitroxymethyl)phenol, the latter leading to (hydroxymethyl)phenol and the glutathione-spacer conjugate. This metabolic pathway takes place in its entirety only in the cytosolic fraction of the tissues tested and in intact human colon cancer cells, perhaps reflecting exposure to the cytosolic glutathione S-transferase, which catalyzes the formation of the spacer-glutathione conjugate. Thus, the three positional isomers of NO-ASA differ in their metabolism and these differences correlate with their differential effects on cancer cell growth, underscoring the importance of positional isomerism in modulating drug effects. PMID- 15528453 TI - Positional isomerism markedly affects the growth inhibition of colon cancer cells by nitric oxide-donating aspirin in vitro and in vivo. AB - NO-donating aspirin (NO-ASA), a novel pharmacological agent currently undergoing clinical testing, consists of ASA to which a nitrate group is covalently linked via a spacer molecule. We synthesized the three positional isomers of NO-ASA with respect to the -CH(2)ONO(2) group (ortho, meta, and para) and examined whether this isomerism affects the biological activity of NO-ASA on HT-29 human colon cancer cells. The ortho- and para-isomers showed similar IC(50) values (1-5 microM) for cell growth inhibition over 72 h, whereas the IC(50) of the meta isomer was 200 to 500 microM. The ortho- and para-isomers inhibited cell proliferation more potently than the meta-isomer. All three induced apoptosis but the ortho- and para-isomers also induced atypical cells (they maintain their shape but have diminished or absent nuclear material). Treatment for 3 weeks of Min (Apc(min)(/+)) mice, a model of intestinal cancer, with equimolar amounts of meta- and para-NO-ASA decreased the number of tumors in the small intestine by 36 and 59% (P < 0.01), respectively, compared with vehicle-treated controls, thus confirming their in vitro differences in potency. A structure-activity study of the three isomers revealed that substituting an aliphatic for the aromatic spacer or removing the -ONO(2) group profoundly diminished NO-ASA's ability to inhibit cell growth, whereas removal of the acetyl group on the ASA moiety did not affect cell growth inhibition. Thus, positional isomerism is critical for the pharmacological properties of NO-ASA against colon cancer and it should be taken into consideration in rational drug design. PMID- 15528454 TI - Bacterial communication ("quorum sensing") via ligands and receptors: a novel pharmacologic target for the design of antibiotic drugs. AB - The purpose of the present Perspectives is to present a synopsis of the literature on bacterial "quorum sensing" as a background for the proposal that interference with this communication system offers potential targets for the design of novel antibiotic drugs. Quorum sensing is the recently discovered chemical communication system among bacteria (both Gram-positive and -negative). It is vital for intra- and interbacterial gene regulation and for keeping bacterial colonies ("biofilms") intact, allowing resident bacteria to assume specialized roles that contribute to enhanced survival of the group. There are several processes involved in quorum sensing that are familiar to pharmacologists; i.e., specific signaling molecules bind to and activate receptors that transduce the quorum-sensing signal into intracellular second messenger responses. We highlight herein the similarity between quorum-sensing communication to ligand-receptor interactions, suggesting that inhibitor drugs could be designed using current standard pharmacologic principles. Such drugs would have novel mechanisms of action and might therefore be more effective against antibiotic-resistant strains of bacteria. PMID- 15528455 TI - Increased expression of hepatocyte growth factor in cerebrospinal fluid and intracranial artery in moyamoya disease. AB - BACKGROUND AND PURPOSE: The etiology of moyamoya disease still remains unknown. This study was aimed to explore the role of hepatocyte growth factor (HGF), a strong inducer of angiogenesis, in development of moyamoya disease. METHODS: We studied cerebrospinal fluid (CSF) from 39 patients with moyamoya disease (24 children and 15 adults), 6 control patients with cervical spondylosis, and 7 control patients with internal carotid artery occlusion. CSF level of HGF was determined by enzyme-linked immunosorbent assay technique. We also evaluated the distribution of HGF and its cellular receptor c-Met in the carotid fork obtained from 2 patients with moyamoya disease and 2 control patients. RESULTS: CSF level of HGF was 408.2+/-201.6 pg/mL and 443.2+/-193.5 pg/mL in patients with cervical spondylosis and internal carotid artery occlusion, respectively (mean+/-SD). On the other hand, CSF level of HGF was 820.3+/-319.0 pg/mL in patients with moyamoya disease, being significantly higher than those in 2 control groups (P<0.01). Both HGF and c-Met were widely distributed in the media and thickened intima of the carotid fork in patients with moyamoya disease but not in control patients. CONCLUSIONS: This study revealed that HGF is densely found in the carotid fork, and its CSF level is markedly elevated in moyamoya disease, suggesting that HGF may be a key protein for pathogenesis of moyamoya disease. PMID- 15528457 TI - Absence of alpha-1 antitrypsin deficiency alleles (S and Z) in Japanese and Korean patients with aneurysmal subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: A possible association has been proposed for the formation of intracranial aneurysm (IA) and deficiency alleles (S and Z) of the alpha1-antitrypsin (AAT) gene. We extensively screened this gene in Japanese and Korean patients with aneurysmal subarachnoid hemorrhage. METHODS: Seven allelic variants, including S and Z alleles, were genotyped by direct sequencing of genomic DNA obtained from 195 and 189 ruptured IA patients and 195 and 94 controls in Japanese and Koreans, respectively. The haplotype in phase-unknown samples was constructed with the expectation-maximization method. Differences in allelic frequencies between patients and controls were evaluated by Fisher exact test. RESULTS: No significant differences in allelic frequencies were observed at all 7 variants between ruptured IA patients and controls. We could not detect the S and Z alleles of the AAT gene in Japanese and Korean populations. CONCLUSIONS: AAT deficiency may not be a common genetic risk factor for aneurysmal subarachnoid hemorrhage in Japanese and Koreans. PMID- 15528459 TI - Poststroke "pushing": natural history and relationship to motor and functional recovery. AB - BACKGROUND AND PURPOSE: Patients with hemiparetic stroke have impaired balance control. Some patients ("pushers") are resistant to accepting weight on and actively "push" away from the nonparetic side. This research identified pushers from stroke patients with moderate to severe hemiparesis and examined longitudinal changes in symptoms, level of impairment, and functional independence. METHODS: Prospective sample of hemiparetic stroke patients (n=65) located in Toronto, Canada. Detailed clinical assessments were performed within 10 days postonset, at 6 weeks, and at 3 months. RESULTS: At 1 week after stroke, 63% of patients demonstrated features of pushing. In 62% of pushers, symptoms resolved by 6 weeks, whereas in 21%, pushing symptoms persisted at 3 months. Motor recovery and functional abilities at 3 months were significantly lower among the pushers compared with the nonpushers. Pushers also had a significantly longer hospital length of stay (89 days versus 57 days). It is noteworthy that motor and functional recovery improved significantly over the 3-month study period for both pushers and nonpushers. CONCLUSIONS: Identification of stroke patients with pushing symptoms has prognostic implications for recovery. In light of this potential recovery, rehabilitation specialists need to refine treatment approaches for the pushers to further improve functional outcome. PMID- 15528460 TI - Carotid intima-media thickness and plaque characteristics as a risk factor for stroke in Japanese elderly men. AB - BACKGROUND AND PURPOSE: Few cohort studies have examined the association of carotid intima-media thickness (IMT) and plaque characteristics with the risk of stroke in apparently healthy persons. We examined the relationship of carotid IMT and the surface, morphology, and calcification of carotid plaques with the incidence of stroke among Japanese men. METHODS: Carotid IMT and plaque were evaluated bilaterally with ultrasonography in 1289 men aged 60 to 74 years without a previous stroke or coronary heart disease. In this cohort, the subsequent incidence of stroke was investigated. RESULTS: During the 4.5-year follow-up, 34 strokes occurred. The multivariate-adjusted relative risk (95% CI) for the highest versus lowest quartiles of maximum IMT of the common carotid artery (CCA; > or =1.07 versus < or =0.77 mm) was 3.0 (1.1 to 8.3) for stroke. The combination of CCA and internal carotid artery (ICA) wall thickness was a better predictor of the risk of stroke than was CCA wall thickness alone. Men with a plaque, defined as a focal wall thickness of > or =1.5 mm, in the ICA had a 3-fold higher risk of stroke than those without a plaque, and the plaque surface irregularity further increased the stroke risk. A significant excess risk of stroke was confined to men with an uncalcified plaque. CONCLUSIONS: Increased IMT of the CCA and an uncalcified plaque in the ICA, as assessed by ultrasonography, are risk factors for stroke in elderly Japanese men. PMID- 15528461 TI - White matter hyperintensities: pearls and pitfalls in interpretation of MRI abnormalities. PMID- 15528463 TI - ABCA1 expression in carotid atherosclerotic plaques. AB - BACKGROUND AND PURPOSE: The ATP-binding cassette transporter A1 (ABCA1) facilitates cholesterol efflux from cells, a key process in reverse cholesterol transport. Whereas previous investigations focused on mutations causing impaired ABCA1 function, we assessed the role of ABCA1 in human carotid atherosclerotic disease. METHODS: We compared the mRNA and protein levels of ABCA1, and one of its key regulators, the liver X receptor alpha (LXRalpha), between minimally and grossly atherosclerotic arterial tissue. We established ABCA1 and LXRalpha gene expression by real-time quantitative polymerase chain reaction in 10 control and 18 atherosclerotic specimens. Presence of ABCA1 protein was assessed by immunoblotting. To determine whether differences observed at a local level were reflected in the systemic circulation, we measured ABCA1 mRNA in leukocytes of 10 patients undergoing carotid endarterectomy and 10 controls without phenotypic atherosclerosis. RESULTS: ABCA1 and LXRalpha gene expression were significantly elevated in atherosclerotic plaques (P<0.0001 and 0.03, respectively). The increased mRNA levels of ABCA1 and LXRalpha were correlated in atherosclerotic tissue (r=0.85; P<0.0001). ABCA1 protein expression was significantly reduced in plaques compared with control tissues (P<0.0001). There were no differences in leukocyte ABCA1 mRNA expression (P=0.67). CONCLUSIONS: ABCA1 gene and protein are expressed in minimally atherosclerotic human arteries. Despite significant upregulation of ABCA1 mRNA, possibly mediated via LXRalpha, ABCA1 protein is markedly reduced in advanced carotid atherosclerotic lesions. No differences in leukocyte ABCA1 expression were found, suggesting the plaque microenvironment may contribute to the differential ABCA1 expression. We propose that the decreased level of ABCA1 protein is a key factor in the development of atherosclerotic lesions. PMID- 15528464 TI - Proarrhythmic consequences of a KCNQ1 AKAP-binding domain mutation: computational models of whole cells and heterogeneous tissue. AB - The KCNQ1-G589D gene mutation, associated with a long-QT syndrome, has been shown to disrupt yotiao-mediated targeting of protein kinase A and protein phosphatase 1 to the I(Ks) channel. To investigate how this defect may lead to ventricular arrhythmia during sympathetic stimulation, we use integrative computational models of beta-adrenergic signaling, myocyte excitation-contraction coupling, and action potential propagation in a rabbit ventricular wedge. Paradoxically, we find that the KCNQ1-G589D mutation alone does not prolong the QT interval. But when coupled with beta-adrenergic stimulation in a whole-cell model, the KCNQ1 G589D mutation induced QT prolongation and transient afterdepolarizations, known cellular mechanisms for arrhythmogenesis. These cellular mechanisms amplified tissue heterogeneities in a three-dimensional rabbit ventricular wedge model, elevating transmural dispersion of repolarization and creating other T-wave abnormalities on simulated electrocardiograms. Increasing heart rate protected both single myocyte and the coupled myocardium models from arrhythmic consequences. These findings suggest that the KCNQ1-G589D mutation disrupts a critical link between beta-adrenergic signaling and myocyte electrophysiology, creating both triggers of cardiac arrhythmia and a myocardial substrate vulnerable to such electrical disturbances. PMID- 15528465 TI - Fibulin-5 is a novel binding protein for extracellular superoxide dismutase. AB - The extracellular superoxide dismutase (ecSOD) plays an important role in atherosclerosis and endothelial function by modulating levels of the superoxide anion (O2*-) in the extracellular space. Although heparan sulfate proteoglycan is an important ligand for ecSOD, little is known about other biological binding partners of ecSOD. The goal of this study was to identify novel proteins that interact with ecSOD. A yeast two-hybrid screening of a human aorta cDNA library using ecSOD as bait identified fibulin-5 as a predominant binding protein for ecSOD. Further analysis showed that the binding domain of ecSOD within fibulin-5 mapped to its C-terminal domain. In vitro pulldown assays and coimmunoprecipitation analysis further confirmed that ecSOD interacts with fibulin-5 in vitro and in vivo. Studies using fibulin-5-/- mice indicated that fibulin-5 is required for binding of ecSOD to vascular tissue. Importantly, the decrease in tissue-bound ecSOD levels in aortas from fibulin-5-/- mice was associated with an increase in vascular O2*- levels. Furthermore, immunohistochemical analysis using ApoE-/- mice suggested a codistribution of ecSOD and fibulin-5 in atherosclerotic vessels. In summary, we provide in this study the first evidence that the ecSOD-fibulin-5 interaction is required for ecSOD binding to vascular tissues, thereby regulating vascular O2*- levels. This interaction may represent a novel mechanism for controlling vascular redox state in the extracellular space in various cardiovascular diseases such as atherosclerosis and hypertension in which oxidative stress is increased. PMID- 15528466 TI - Transforming growth factor beta-SMAD2 signaling regulates aortic arch innervation and development. AB - Aortic arch interruptions in humans and animal models are mainly caused by aberrant development of the fourth pharyngeal arch artery. Little is known about the maturation of this vessel during normal and abnormal development, which is the subject of this study. Tgfbeta2 knockout mice that present with fourth artery defects have been associated with defective neural crest cell migration. In this study, we concentrated on pharyngeal arch artery development during developmental days 12.5 to 18.5, focusing on neural crest cell migration using a Wnt1-Cre by R26R neural crest cell reporter mouse. Fourth arch artery maturation was studied with antibodies directed against smooth muscle alpha-actin and neural NCAM-1 and RMO-270. For diminished transforming growth factor beta (TGF-beta) signaling, SMAD2 and fibronectin have been analyzed. Neural crest migration and differentiation into smooth muscle cells is unaltered in mutants, regardless of the cardiovascular defect found; however, innervation of the fourth arch artery is affected. Absent staining for nuclear SMAD2, NCAM-1, and RMO-270 in the fourth artery in mutant coincides with severe defects of this segment. Likewise, fibronectin expression is diminished in these cases. From these data we conclude the following: (1) neural crest cell migration is not a common denominator in cardiovascular defects of Tgfbeta2-/- mice; (2) fourth arch artery maturation is a complex process involving innervation; and (3) TGF-beta2 depletion diminishes SMAD2-signaling in the fourth arch artery and coincides with reduced vascular NCAM-1 expression and neural innervation of this artery. We hypothesize that disturbed maturation of the fourth pharyngeal arch artery, and especially abrogated vascular innervation, will result in fourth arch interruptions. PMID- 15528467 TI - Induction of the MMP-14 gene in macrophages of the atherosclerotic plaque: role of SAF-1 in the induction process. AB - Based on epidemiological and pathological studies, it is becoming increasingly clear that matrix metalloproteinases (MMPs) play an important role in the pathogenesis of atherosclerosis by participating in vascular remodeling, smooth muscle cell migration, and plaque disruption. MMP-14, because of its unique ability to cause pericellular degradation, its broad substrate specificity, its synthesis in an active form, and its ability to activate other matrix metalloproteinases, is recognized as a prominent member of this family. MMP-14 is detected at high levels in the atherosclerotic plaque. To understand the induction mechanism of MMP-14 under atherogenic conditions, we examined its expression pattern in response to oxidized low-density lipoproteins (ox-LDLs) that are believed to play an important role in atherogenesis. We report that in macrophages, ox-LDLs markedly elevate the levels of MMP-14 mRNA and protein. The cis-acting elements supporting this increase were identified to be present within -213 and -1 nucleotides of the MMP-14 promoter. DNase I protection assay revealed, within this region, two major elements, of which one serves as the DNA binding site for SAF-1 transcription factor. Increased binding of SAF-1 to the MMP-14 promoter correlated with the transcriptional upregulation of MMP-14 gene. Furthermore, induction of endogenous MMP-14 gene, MMP-14 promoter driven reporter gene expression and MMP-2 processing activity during overexpression of SAF-1 and coexpression of SAF-1 and MMP-14 in the macrophages present in the atherosclerotic plaque implicate SAF-1 as a key regulator of MMP-14 gene induction in macrophage cells. PMID- 15528468 TI - Xanthine oxidoreductase is an endogenous regulator of cyclooxygenase-2. AB - Xanthine oxidoreductase (XOR) is the enzyme responsible for the final step in purine degradation resulting in the generation of uric acid. Here we have generated mice deficient in XOR. As expected, these animals lack tissue XOR activity and have low to undetectable serum levels of uric acid. Although normal at birth, XOR-/- mice fail to thrive after 10 to 14 days, and most die within the first month. The cause of death appears to be a form of severe renal dysplasia, a phenotype that closely resembles what has been observed previously in cyclooxygenase-2 (COX-2)-deficient mice. We further demonstrate that in the first month of life, a period in which the mouse kidney is undergoing rapid maturation and remodeling, wild-type mice exhibit an approximately 30-fold increase in renal XOR activity, with a corresponding induction of COX-2 expression. In contrast, during this same period, XOR-/- animals fail to augment renal COX-2 expression. Finally, we show that in vitro and in vivo, uric acid can stimulate basal COX-2 expression. These results demonstrate that XOR activity is an endogenous physiological regulator of COX-2 expression and thereby provide insight into previous epidemiological evidence linking elevated serum uric levels with systemic hypertension and increased mortality from cardiovascular diseases. In addition, these results suggest a novel molecular link between cellular injury and the inflammatory response. PMID- 15528469 TI - Hypertension-linked mutation in the adducin alpha-subunit leads to higher AP2-mu2 phosphorylation and impaired Na+,K+-ATPase trafficking in response to GPCR signals and intracellular sodium. AB - Alpha-adducin polymorphism in humans is associated with abnormal renal sodium handling and high blood pressure. The mechanisms by which mutations in adducin affect the renal set point for sodium excretion are not known. Decreases in Na+,K+-ATPase activity attributable to endocytosis of active units in renal tubule cells by dopamine regulates sodium excretion during high-salt diet. Milan rats carrying the hypertensive adducin phenotype have a higher renal tubule Na+,K+-ATPase activity, and their Na+,K+-ATPase molecules do not undergo endocytosis in response to dopamine as do those of the normotensive strain. Dopamine fails to promote the interaction between adaptins and the Na+,K+-ATPase because of adaptin-mu2 subunit hyperphosphorylation. Expression of the hypertensive rat or human variant of adducin into normal renal epithelial cells recreates the hypertensive phenotype with higher Na+,K+-ATPase activity, mu2 subunit hyperphosphorylation, and impaired Na+,K+-ATPase endocytosis. Thus, increased renal Na+,K+-ATPase activity and altered sodium reabsorption in certain forms of hypertension could be attributed to a mutant form of adducin that impairs the dynamic regulation of renal Na+,K+-ATPase endocytosis in response to natriuretic signals. PMID- 15528470 TI - Retardation of atherosclerosis by overexpression of catalase or both Cu/Zn superoxide dismutase and catalase in mice lacking apolipoprotein E. AB - Oxidative stress has been suggested to potentiate atherogenesis. However, studies that have investigated the effect of antioxidants on atherosclerosis showed inconsistent results, ie, atherosclerosis was either retarded or not changed by dietary antioxidants. This report directly examined the effect of overexpressing Cu/Zn-superoxide dismutase (Cu/Zn-SOD) and/or catalase on atherosclerosis and lipid peroxidation in mice lacking apolipoprotein E (ApoE-/-). Based on lipid staining of the en face of the aorta tree and the serial sections of the proximal aorta, ApoE-/- mice overexpressing catalase or both Cu/Zn-SOD and catalase had smaller and relatively early stages of atherosclerotic lesions (eg, foam cells and free lipids) when compared with ApoE-/- mice, who developed more advanced lesions (eg, fibrous caps and acellular areas). In addition, the retarded development of atherosclerosis was correlated with a reduced F2-isoprostanes in the plasma and aortas in ApoE-/- mice overexpressing catalase or both Cu/Zn-SOD and catalase. In contrast, the levels of F2-isoprostanes and atherosclerosis in the ApoE-/- mice overexpressing Cu/Zn-SOD alone were comparable to ApoE-/- control mice. These observations implied that endogenously produced hydrogen peroxide, but not superoxide anions, contributed to the formation of oxidized lipids and the development of atherosclerosis in ApoE-/- mice. PMID- 15528471 TI - Alterations in myofilament function contribute to left ventricular dysfunction in pigs early after myocardial infarction. AB - Myocardial infarction (MI) initiates cardiac remodeling, depresses pump function, and predisposes to heart failure. This study was designed to identify early alterations in Ca2+ handling and myofilament proteins, which may contribute to contractile dysfunction and reduced beta-adrenergic responsiveness in postinfarct remodeled myocardium. Protein composition and contractile function of skinned cardiomyocytes were studied in remote, noninfarcted left ventricular (LV) subendocardium from pigs 3 weeks after MI caused by permanent left circumflex artery (LCx) ligation and in sham-operated pigs. LCx ligation induced a 19% increase in LV weight, a 69% increase in LV end-diastolic area, and a decrease in ejection fraction from 54+/-5% to 35+/-4% (all P<0.05), whereas cardiac responsiveness to exercise-induced increases in circulating noradrenaline levels was blunted. Endogenous protein kinase A (PKA) was significantly reduced in remote myocardium of MI animals, and a negative correlation (R=0.62; P<0.05) was found between cAMP levels and LV weight-to-body weight ratio. Furthermore, SERCA2a expression was 23% lower after MI compared with sham. Maximal isometric force generated by isolated skinned myocytes was significantly lower after MI than in sham (15.4+/-1.5 versus 19.2+/-0.9 kN/m2; P<0.05), which might be attributable to a small degree of troponin I (TnI) degradation observed in remodeled postinfarct myocardium. An increase in Ca2+ sensitivity of force (pCa50) was observed after MI compared with sham (DeltapCa50=0.17), which was abolished by incubating myocytes with exogenous PKA, indicating that the increased Ca2+ sensitivity resulted from reduced TnI phosphorylation. In conclusion, remodeling of noninfarcted pig myocardium is associated with decreased SERCA2a and myofilament function, which may contribute to depressed LV function. The full text of this article is available online at http://circres.ahajournals.org. PMID- 15528472 TI - Crucial role of the CCL2/CCR2 axis in neointimal hyperplasia after arterial injury in hyperlipidemic mice involves early monocyte recruitment and CCL2 presentation on platelets. AB - Monocyte chemoattractant protein-1 (also known as CC chemokine ligand 2 [CCL2]) and its receptor CC chemokine receptor 2 (CCR2) play a central role in the inflammatory response and neointimal formation after vascular injury. In the context of hyperlipidemia, this appears to involve neointimal monocyte infiltration. Hence, we investigated the function of the CCL2/CCR2 axis in early monocyte recruitment to injured arteries. Wire-induced injury of the carotid artery in apoE-/- mice caused a rapid increase of JE/CCL2 protein in the vessel wall peaking at 24 hours after injury, whereas serum JE/CCL2 was increased solely at 6 hours and blood cell-associated levels were unaltered, as demonstrated by enzyme-linked immunosorbent assay. Immunohistochemistry revealed intense staining for JE/CCL2 in smooth muscle cells (SMCs) and in association with platelets adherent to the denuded vessel wall 24 hours after injury. In vitro, exogenous or SMC-derived JE/CCL2 binds to the platelet surface and triggers monocyte arrest on adherent platelets but not on SMCs in flow assays. Accordingly, monocyte arrest in ex vivo perfused apoE-/- carotid arteries isolated 24 hours after injury was profoundly inhibited by pretreatment with a JE/CCL2 antibody. In CCR2-/-/apoE-/- mice, neointimal plaque area was reduced by 47% compared with CCR2+/+/apoE-/- mice. Moreover, CCR2 deletion markedly decreased neointimal macrophage content while expanding SMC content. Vascular JE/CCL2 expressed by SMCs and immobilized by adherent platelets after endothelial denudation is crucial for mediating early monocyte recruitment to injured arteries in hyperlipidemic mice. This mechanism may explain reduced neointimal macrophage infiltration and lesion formation in CCR2-deficient apoE-/- mice. PMID- 15528473 TI - Dual effect of angiotensin-converting enzyme inhibition on angiogenesis in type 1 diabetic mice. AB - OBJECTIVE: We analyzed the beneficial therapeutic effect of angiotensin converting enzyme inhibitor (ACEI) on both retinal and hind limb neovascularization in diabetic mice. METHODS AND RESULTS: Diabetic mice (streptozotocin, 40 mg/kg) were treated with or without ACEI (Perindopril, 3 mg/kg per day) or AT1 receptor blocker (Candesartan, 20 mg/kg) for 4 months. Hind limb ischemia was then induced by right femoral artery ligature for 1 additional month. In the ischemic leg, angiographic score, capillary density, and foot perfusion were increased by 2.7, 2.0-fold, and 1.6-fold, respectively, in ACEI treated diabetic mice compared with untreated diabetic animals (P<0.01). ACEI also raised vascular endothelial growth factor (VEGF) protein level by 1.4-fold in ischemic diabetic leg. This ACEI pro-angiogenic effect was totally blunted in diabetic bradykinin B2 receptor-deficient animals, suggesting that it was mediated by the bradykinin pathway. In the diabetic retina, angiotensinogen and ACE mRNA levels were increased by 2.8-fold and 4.1-fold, respectively (P<0.01 versus nondiabetic mice), highlighting a local activation of renin-angiotensin system. Diabetes also raised VEGF protein level by 1.5-fold (P<0.05 versus nondiabetic mice). Treatments with ACEI and AT1 receptor blocker hampered diabetes-induced VEGF upregulation and retinal neovascularization. CONCLUSIONS: ACE inhibition improved neovascularization in the diabetic ischemic leg through activation of bradykinin signaling, whereas it reduced vessel growth in the diabetic retina through inhibition of overacting Ang II pathway. PMID- 15528474 TI - Ceramide-induced impairment of endothelial function is prevented by CuZn superoxide dismutase overexpression. AB - OBJECTIVE: Ceramide is an important intracellular second messenger that may also increase superoxide. The goal of this study was to determine whether overexpression of CuZn superoxide dismutase (SOD) protects against ceramide induced increases in vascular superoxide and endothelial dysfunction. METHODS AND RESULTS: Carotid arteries from CuZnSOD-transgenic (CuZnSOD-Tg) and nontransgenic littermates were examined in vitro. Immunohistochemistry confirmed that CuZnSOD protein was greater in carotid artery from CuZnSOD-Tg compared with nontransgenic mice. Ceramide (N-acetyl-d-sphingosine; 1 and 10 micromol/L) produced concentration-dependent impairment (P<0.05) of vasorelaxation in response to the endothelium-dependent agonist acetylcholine (ACh) in nontransgenic mice. For example, 100 micromol/L ACh relaxed arteries from nontransgenic mice by 96+/-4% and 52+/-5% in the presence of vehicle and 10 micromol/L ceramide, respectively. In contrast, ceramide (1 or 10 micromol/L) had no effect (P>0.05) on responses of carotid artery to ACh in CuZnSOD-Tg mice. Ceramide had no effect on nitroprusside or papaverine-induced relaxation in CuZnSOD-Tg or nontransgenic mice. Ceramide increased superoxide in arteries from nontransgenic vessels, and this effect was prevented by polyethyleneglycol-SOD (50 U/mL) or overexpression of CuZnSOD. CONCLUSIONS: These results suggest that ceramide-induced increases in superoxide impair endothelium-dependent relaxation, and that select overexpression of the CuZn isoform of SOD prevents ceramide-induced oxidative stress in vessels. PMID- 15528475 TI - Quantitative evaluation of carotid plaque composition by in vivo MRI. AB - OBJECTIVE: This study evaluates the ability of MRI to quantify all major carotid atherosclerotic plaque components in vivo. METHODS AND RESULTS: Thirty-one subjects scheduled for carotid endarterectomy were imaged with a 1.5T scanner using time-of-flight-, T1-, proton density-, and T2-weighted images. A total of 214 MR imaging locations were matched to corresponding histology sections. For MRI and histology, area measurements of the major plaque components such as lipid rich/necrotic core (LR/NC), calcification, loose matrix, and dense (fibrous) tissue were recorded as percentages of the total wall area. Intraclass correlation coefficients (ICCs) were computed to determine intrareader and inter reader reproducibility. MRI measurements of plaque composition were statistically equivalent to those of histology for the LR/NC (23.7 versus 20.3%; P=0.1), loose matrix (5.1 versus 6.3%; P=0.1), and dense (fibrous) tissue (66.3% versus 64%; P=0.4). Calcification differed significantly when measured as a percentage of wall area (9.4 versus 5%; P<0.001). Intrareader and inter-reader reproducibility was good to excellent for all tissue components, with ICCs ranging from 0.73 to 0.95. CONCLUSIONS: MRI-based tissue quantification is accurate and reproducible. This application can be used in therapeutic clinical trials and in prospective longitudinal studies to examine carotid atherosclerotic plaque progression and regression. PMID- 15528476 TI - Sulfatides activate platelets through P-selectin and enhance platelet and platelet-leukocyte aggregation. AB - OBJECTIVE: Sulfatides are sulfated glycosphingolipids present on the surface of a variety of cells; however, their exact physiological function is not known. Recently, we have shown that the inhibition of sulfatide-P-selectin interactions leads to disaggregation of platelet aggregates. METHODS AND RESULTS: In this study, we show that sulfatides activated platelets as they increased activation of GPIIb/IIIa (PAC-1 epitope) and expression of P-selectin on the platelet surface. Furthermore, sulfatides aggregated washed platelets in a dose-dependent manner and enhanced platelet aggregation in platelet-rich plasma. Previous activation of platelets was necessary for this effect. Monoclonal anti-P-selectin antibodies inhibited not only sulfatide-induced PAC-1 binding to platelets but also sulfatide-induced platelet aggregation, suggesting that sulfatides activate platelet GPIIb/IIIa via signaling through P-selectin. The proaggegatory effect of sulfatides was also observed in an ex vivo thrombosis model using whole blood and pulsatile flow at 37 degrees C. In this model, sulfatides significantly enhanced platelet aggregation and the formation of platelet-leukocyte aggregates. CONCLUSIONS: We show that sulfatide-P-selectin interactions lead to subsequent platelet activation and P-selectin expression, forming a positive feedback loop that can potentiate formation of stable platelet aggregates. In addition, sulfatides enhance the aggregation of platelet-leukocyte aggregates. These mechanisms may play a significant role in hemostasis and thrombosis. PMID- 15528477 TI - Hyperhomocysteinemia is inversely related with left ventricular ejection fraction and predicts cardiovascular mortality in high-risk coronary artery disease hypertensives. AB - OBJECTIVE: The purpose of this study was to investigate the relationship of plasma homocysteine (tHcy) levels with coronary artery disease (CAD) and left ventricular ejection fraction (LVEF) in high-risk patients undergoing coronary angiography for suspected CAD. METHODS AND RESULTS: In 936 consecutive patients, we measured LVEF, tHcy, folate levels, and quantified CAD with a modified Duke Index score. We also genotyped patients at the methylen-tetrahydrofolate reductase 677C-->T polymorphism. Hyperhomocysteinemia (HHcy) was defined as tHcy levels > or =15.46 micromol/L; total and cardiovascular mortality was assessed at follow-up that lasted 43 months (median). CAD was confirmed in 75% of patients and ruled out in the rest (non-CAD group). No relationship of HHcy with either arterial hypertension or the CAD score was found. In contrast, there was a significant inverse relationship of tHcy with LVEF in arterial hypertensive but not in normotensive patients, regardless of previous myocardial infarction. At logistic regression, HHcy was the strongest predictor (P=0.001) of a low (<40%) LVEF, followed by type 2 diabetes mellitus and cigarette smoking. At follow-up, HHcy significantly predicted cardiovascular mortality but only in the arterial hypertension subgroup. CONCLUSIONS: In arterial hypertensive but not in normotensive patients, HHcy predicts cardiovascular mortality and a low LVEF, independent of CAD and history of myocardial infarction. PMID- 15528478 TI - MRI of atherothrombosis associated with plaque rupture. AB - OBJECTIVE: MRI would be a valuable noninvasive diagnostic tool to study plaque associated thrombi. We defined the imaging characteristics of these thrombi, composed primarily of platelets and fibrin, and distinguished them clearly from the vessel lumen and underlying atherosclerotic plaque in an animal model of plaque rupture. METHODS AND RESULTS: After triggering plaque rupture in New Zealand White male rabbits, segments of infrarenal aorta containing either red or white thrombi were fixed in formalin. Compared with postmortem red cell-rich thrombi, atherothrombi yielded complex magnetic resonance images with intermediate signal intensity in standard T1- and T2-weighted imaging sequences and were often difficult to distinguish from the aortic wall. Diffusion-weighted imaging sequences revealed restricted diffusion of the atherothrombus relative to the vessel wall and provided excellent contrast. The apparent diffusion coefficient of the thrombus is 1.0x10(-3) mm2/s, compared with 1.5x10(-3) mm2/s in tissue. Similar results were obtained using purified aggregated platelets. CONCLUSIONS: We present the first detailed description of the MRI appearance of plaque rupture-associated thrombosis in histologically validated platelet-rich thrombi. Diffusion-weighted imaging provided the best distinction between thrombus and vessel wall and has potential application for the noninvasive in vivo detection of atherothrombosis. PMID- 15528479 TI - Scavenger receptor class B type I mediates the selective uptake of high-density lipoprotein-associated cholesteryl ester by the liver in mice. AB - OBJECTIVE: High-density lipoprotein (HDL) cholesteryl esters (CE) are taken up by liver and adrenals selectively, ie, independent from particle internalization. Class B type I scavenger receptor (SR-BI) mediates this uptake in vitro. The role of SR-BI in HDL metabolism was explored in mice. METHODS AND RESULTS: Mice with a mutation in the SR-BI gene (SR-BI KO) and wild-type (WT) littermates were used. Mutants had increased HDL cholesterol. HDL was labeled with 125I (protein) and [3H] (CE). After HDL injection, blood samples were drawn and finally the mice were euthanized. In WT, the plasma decay of HDL-associated [3H] is faster compared with 125I and this represents whole-body selective CE uptake. In SR-BI KO, the decay of both tracers is similar, yielding no selective CE removal. In WT liver and adrenals, uptake of [3H] is higher than 125I, showing selective uptake. In SR-BI KO, liver uptake of [3H] and 125I are similar, proposing no selective HDL CE uptake. In SR-BI KO adrenals, selective uptake is reduced; however, even in the absence of SR-BI, this uptake is detected using WT-HDL. CONCLUSIONS: SR-BI mediates selective uptake of HDL CE by the liver. In adrenals, an alternative mechanism or mechanisms can play a role in selective CE uptake. PMID- 15528480 TI - Phenotype of heterozygotes for low-density lipoprotein receptor mutations identified in different background populations. AB - BACKGROUND: The effect of mutations on phenotype is often overestimated because of ascertainment bias. We determined the effect of background population on cholesterol phenotype associated with specific mutations in the low-density lipoprotein (LDL) receptor and the relative importance of background population and type of mutation (LDL receptor [LDLR] or APOB R3500Q) for cholesterol phenotype. METHODS AND RESULTS: We studied 9255 individuals from the general population, 948 patients with ischemic heart disease (IHD), and 63 patients with clinical familial hypercholesterolemia (FH) for 3 common LDL receptor mutations. Average increase in cholesterol in LDL receptor heterozygotes identified in the general population or among patients with IHD or FH compared with noncarriers was 2.9 mmol/L, 4.1 mmol/L, and 4.9 mmol/L, respectively (P=0.02). Background population and type of mutation determined cholesterol phenotype; average increase in LDL cholesterol from carriers in the general population to carriers with clinical FH was 1.6 mmol/L (P=0.03). The average increase for carriers of LDLR mutations compared with carriers of APOB R3500Q was 1.2 mmol/L (P=0.05). CONCLUSIONS: The phenotype associated with a given mutation should not be determined in patients, but rather in unselected individuals in the general population. PMID- 15528481 TI - Genotypic effect of the -565C>T polymorphism in the ABCA1 gene promoter on ABCA1 expression and severity of atherosclerosis. AB - OBJECTIVE: Loss-of-function mutations of the ATP-binding cassette transporter A1 (ABCA1) gene cause Tangier disease, a rare genetic disorder with accumulation of lipid-laden macrophages and increased risk of atherosclerosis. Common variants of this gene may be a genetic factor for atherosclerosis in the general population. This study was performed to test the reported association between the -565C>T polymorphism and atherosclerosis severity and to investigate whether this variant per se had an effect on promoter activity of the ABCA1 gene. METHODS AND RESULTS: A cohort of patients with coronary atherosclerosis were genotyped for the -565C>T polymorphism. Logistic regression analyses showed that homozygotes of the -565T allele had greatest mean number of diseased coronary arteries, particular in nonsmokers. Real-time reverse-transcriptase polymerase chain reaction showed that in atherosclerotic plaques removed from patients undergoing endarteretomy, ABCA1 expression levels were lowest in those who had the T/T genotype and highest in those of the C/C genotype. Transfection and reporter assays demonstrated that in cultured macrophages, the -565T allelic promoter had a lower activity in driving gene expression than the -565C allelic promoter. Electrophoretic mobility shift assays displayed differential binding of nuclear proteins to the 2 alleles. CONCLUSIONS: These results indicate that the -565C>T polymorphism has an allele specific effect on ABCA1 gene expression and provide further evidence of a genotypic effect on coronary atherosclerosis severity. The study showed that the ABCA1 gene -565C>T polymorphism was associated with severity of coronary atherosclerosis in a cohort of patients from Southern England and that this sequence variant per se had an effect on promoter activity of the ABCA1 gene. The data support the notion that common ABCA1 gene variants can contribute to interindividual variability in atherosclerosis susceptibility and severity. PMID- 15528482 TI - Reduced in vivo aortic uptake of radiolabeled oxidation-specific antibodies reflects changes in plaque composition consistent with plaque stabilization. AB - OBJECTIVE: Labeled oxidation-specific antibodies (Ox-AB) detect, quantify, and noninvasively image lipid-rich atherosclerotic lesions. However, it is unknown whether Ox-AB detect plaque stabilization. METHODS AND RESULTS: The aortic uptake of intravenously injected 125I-MDA2 (Ox-AB to malondialdehyde [MDA]-low-density lipoprotein [LDL]) was quantitated in: (1) LDL receptor-/- mice with established atherosclerosis continued on Western diet (Progression) or switched to chow (Regression) or chow+vitamins E and C (Regression-VIT) for 6 months; and (2) Watanabe rabbits (3- to 57-months old) with naturally evolved atherosclerotic lesions. In mice, the Progression group had more extensive atherosclerosis, higher 125I-MDA2 uptake, high concordance of Sudan (lipid)-staining and 125I-MDA2 uptake, and stronger oxidized LDL (OxLDL) and macrophage immunostaining than both Regression groups. In contrast, the Regression groups showed Sudan-positive lesions with focally diminished 125I-MDA2 uptake, which coincided with reduced OxLDL and macrophages but more smooth muscle cells (SMCs) and collagen. In rabbits, areas of increased 125I-MDA2 uptake were associated with high Sudan concordance and strong immunostaining for OxLDL and macrophages. Interestingly, advanced lesions with focally diminished 125I-MDA2 uptake showed stronger immunostaining for SMCs and collagen, particularly at the fibrous cap. CONCLUSIONS: Ox-AB uptake is focally diminished in plaques displaying accepted features of plaque stability. Imaging techniques to detect the presence and depletion of OxLDL may be useful in assessing plaque stabilization. PMID- 15528484 TI - Measurement of platelet collagen receptor density in human subjects. PMID- 15528483 TI - The antiinflammatory effects of purple grape juice consumption in subjects with stable coronary artery disease. PMID- 15528485 TI - Gender dependent association of thrombospondin-4 A387P polymorphism with myocardial infarction. PMID- 15528486 TI - Mouse models of vein grafts. PMID- 15528487 TI - Noninvasive phenotypes of atherosclerosis. PMID- 15528488 TI - Estrogens, progestins, and atherosclerosis. PMID- 15528489 TI - Endothelial progenitor cells at work: not mature yet, but already stress resistant. PMID- 15528490 TI - Pseudomonas aeruginosa degrades pulmonary surfactant and increases conversion in vitro. AB - Although it is known that surfactant lipids and proteins are altered in patients with Pseudomonas aeruginosa infections, the mechanisms and implications of these alterations are not clear. In this study, the effects of P. aeruginosa on the surfactant large aggregate fraction were examined using an in vitro surface area cycling model. Large aggregates were isolated from porcine bronchoalveolar lavage fluid and incubated with supernatants from P. aeruginosa cultures (PAO1, parent strain; PAO1-A1, lasA-negative mutant; PAO1-B1, elastase-negative mutant) or purified elastase. Amounts of surfactant protein (SP)-A and SP-B, phospholipid content, and large aggregate conversion were assessed. In addition, lipid degradation was assessed by incubating a mixture of radiolabeled phospholipids with P. aeruginosa supernatants. The results demonstrated that SP-A was degraded by PAO1 and PAO1-A1 supernatants, and by purified elastase. SP-B was degraded by PAO1 and PAO1-B1 supernatants, but not by elastase. P. aeruginosa supernatants degraded phospholipids, a process inhibited by ZnCl(2). P. aeruginosa supernatants and elastase increased conversion. The data suggest that protein degradation facilitates increased conversion, and that phospholipid degradation and conversion enhance degradation of surfactant proteins. In conclusion, P. aeruginosa secretes multiple virulence factors that cooperate to result in degradation of surfactant components and alteration of large aggregate conversion. PMID- 15528491 TI - Matrix metalloproteinase/epidermal growth factor receptor/mitogen-activated protein kinase signaling regulate fra-1 induction by cigarette smoke in lung epithelial cells. AB - Exposure to cigarette smoke (CS) can lead to the development of lung cancer, but the molecular mechanisms underlying this process remain unclear. Given that activator protein 1 (AP-1) regulates genes involved in both physiologic and pathophysiologic processes, we have investigated the effects of CS on Jun and Fos family member expression and regulation using a nonmalignant human bronchial epithelial cell line, 1HAEo. Exposure to CS caused a marked upregulation of c Jun, c-Fos, and Fra-1, but not of Fra-2, Jun-B, and Jun-D expression. Because Fra 1 is overexpressed in various tumors and upregulates genes associated with tumor progression, we further elucidated the mechanisms that control CS-stimulated fra 1 induction. CS stimulated fra-1 induction primarily at the transcriptional level. However, epidermal growth factor receptor (EGFR)-specific inhibitor, AG1478, completely suppressed CS-stimulated fra-1 expression. Similarly, the specific inhibitors of extracellular signal-regulated kinase (ERK), c-Jun NH2 terminal kinase (JNK), and p38 kinase signaling markedly suppressed fra-1 induction. Consistent with this finding, AG1478 blocked CS-stimulated ERK, JNK, and p38 phosphorylation. These results suggest that EGFR-activated multiple kinase signaling is essential for fra-1 induction. Furthermore, treatment of cells with GM6001, which inhibits matrix metalloproteinase activity, significantly suppressed CS-stimulated EGF shedding, EGFR and ERK kinase phosphorylation, and subsequent fra-1 induction. Collectively, our findings indicate an obligatory role for metalloproteinase-EGFR-mediated mitogen-activated protein kinase signaling in controlling CS-induced fra-1 expression. PMID- 15528492 TI - Characterization of the nodularin synthetase gene cluster and proposed theory of the evolution of cyanobacterial hepatotoxins. AB - Nodularia spumigena is a bloom-forming cyanobacterium which produces the hepatotoxin nodularin. The complete gene cluster encoding the enzymatic machinery required for the biosynthesis of nodularin in N. spumigena strain NSOR10 was sequenced and characterized. The 48-kb gene cluster consists of nine open reading frames (ORFs), ndaA to ndaI, which are transcribed from a bidirectional regulatory promoter region and encode nonribosomal peptide synthetase modules, polyketide synthase modules, and tailoring enzymes. The ORFs flanking the nda gene cluster in the genome of N. spumigena strain NSOR10 were identified, and one of them was found to encode a protein with homology to previously characterized transposases. Putative transposases are also associated with the structurally related microcystin synthetase (mcy) gene clusters derived from three cyanobacterial strains, indicating a possible mechanism for the distribution of these biosynthetic gene clusters between various cyanobacterial genera. We propose an alternative hypothesis for hepatotoxin evolution in cyanobacteria based on the results of comparative and phylogenetic analyses of the nda and mcy gene clusters. These analyses suggested that nodularin synthetase evolved from a microcystin synthetase progenitor. The identification of the nodularin biosynthetic gene cluster and evolution of hepatotoxicity in cyanobacteria reported in this study may be valuable for future studies on toxic cyanobacterial bloom formation. In addition, an appreciation of the natural evolution of nonribosomal biosynthetic pathways will be vital for future combinatorial engineering and rational design of novel metabolites and pharmaceuticals. PMID- 15528493 TI - Dynamics of a pasture soil microbial community after deposition of cattle urine amended with [13C]urea. AB - Within grazed pastures, urine patches are hot spots of nitrogen turnover, since dietary N surpluses are excreted mainly as urea in the urine. This short-term experiment investigated 13C uptake in microbial lipids after simulated deposition of cattle urine at 10.0 and 17.1 g of urea C m(-2). Confined field plots without or with cattle urine amendment were sampled after 4 and 14 days, and soil from 0- to 5-cm and 10- to 20-cm depths was analyzed for content and composition of phospholipid fatty acids (PLFAs) and for the distribution of urea-derived 13C among individual PLFAs. Carbon dioxide emissions were quantified, and the contributions derived from urea were assessed. Initial changes in PLFA composition were greater at the lower level of urea, as revealed by a principal component analysis. At the higher urea level, osmotic stress was indicated by the dynamics of cyclopropane fatty acids and branched-chain fatty acids. Incorporation of 13C from [13C]urea was low but significant, and the largest amounts of urea-derived C were found in common fatty acids (i.e., 16:0, 16:1omega7c, and 18:1omega7) that would be consistent with growth of typical NH4(+)-oxidizing (Nitrosomonas) and NO2(-)-oxidizing (Nitrobacter) bacteria. Surprisingly, a 20 per thousand depletion of 13C in the cyclopropane fatty acid cy17:0 was observed after 4 days, which was replaced by a 10 to 20 per thousand depletion of that in cy19:0 after 14 days. Possible reasons for this pattern are discussed. Autotrophic nitrifiers could not be implicated in urea hydrolysis to any large extent, but PLFA dynamics and the incorporation of urea-derived 13C in PLFAs indicated a response of nitrifiers which differed between the two urea concentrations. PMID- 15528494 TI - Inactivation of an ABC transporter gene, mcyH, results in loss of microcystin production in the cyanobacterium Microcystis aeruginosa PCC 7806. AB - The cyanobacterium Microcystis aeruginosa is widely known for its production of the potent hepatotoxin microcystin. Microcystin is synthesized nonribosomally by the thiotemplate function of a large, modular enzyme complex encoded within the 55-kb microcystin synthetase (mcy) gene cluster. Also encoded within the mcy gene cluster is a putative ATP binding cassette (ABC) transporter, McyH. This study details the bioinformatic and mutational analyses of McyH and offers functional predictions for the hypothetical protein. The transporter is putatively comprised of two homodimers, each with an N-terminal hydrophobic domain and a C-terminal ATPase. Phylogenetically, McyH was found to cluster with members of the ABC-A1 subgroup of ABC ATPases, suggesting an export function for the protein. Two mcyH null mutant (DeltamcyH) strains were constructed by partial deletion of the mcyH gene. Microcystin production was completely absent in these strains. While the mcyH deletion had no apparent effect on the transcription of other mcy genes, the complete microcystin biosynthesis enzyme complex could not be detected in DeltamcyH mutant strains. Finally, expression levels of McyH in the wild type and in DeltamcyA, DeltamcyB, and DeltamcyH mutants were investigated by using immunoblotting with an anti-McyH antibody. Expression of McyH was found to be reduced in DeltamcyA and DeltamcyB mutants and completely absent in the DeltamcyH mutant. By virtue of its association with the mcy gene cluster and the bioinformatic and experimental data presented in this study, we predict that McyH functions as a microcystin exporter and is, in addition, intimately associated with the microcystin biosynthesis pathway. PMID- 15528495 TI - Highly efficient production of laccase by the basidiomycete Pycnoporus cinnabarinus. AB - An efficient transformation and expression system was developed for the industrially relevant basidiomycete Pycnoporus cinnabarinus. This was used to transform a laccase-deficient monokaryotic strain with the homologous lac1 laccase gene placed under the regulation of its own promoter or that of the SC3 hydrophobin gene or the glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Schizophyllum commune. SC3-driven expression resulted in a maximal laccase activity of 107 nkat ml(-1) in liquid shaken cultures. This value was about 1.4 and 1.6 times higher in the cases of the GPD and lac1 promoters, respectively. lac1-driven expression strongly increased when 25 g of ethanol liter(-1) was added to the medium. Accordingly, laccase activity increased to 1,223 nkat ml( 1). These findings agree with the fact that ethanol induces laccase gene expression in some fungi. Remarkably, lac1 mRNA accumulation and laccase activity also strongly increased in the presence of 25 g of ethanol liter(-1) when lac1 was expressed behind the SC3 or GPD promoter. In the latter case, a maximal laccase activity of 1,393 nkat ml(-1) (i.e., 360 mg liter(-1)) was obtained. Laccase production was further increased in transformants expressing lac1 behind its own promoter or that of GPD by growth in the presence of 40 g of ethanol liter(-1). In this case, maximal activities were 3,900 and 4,660 nkat ml(-1), respectively, corresponding to 1 and 1.2 g of laccase per liter and thus representing the highest laccase activities reported for recombinant fungal strains. These results suggest that P. cinnabarinus may be a host of choice for the production of other proteins as well. PMID- 15528496 TI - Fatty acid production from amino acids and alpha-keto acids by Brevibacterium linens BL2. AB - Low concentrations of branched-chain fatty acids, such as isobutyric and isovaleric acids, develop during the ripening of hard cheeses and contribute to the beneficial flavor profile. Catabolism of amino acids, such as branched-chain amino acids, by bacteria via aminotransferase reactions and alpha-keto acids is one mechanism to generate these flavorful compounds; however, metabolism of alpha keto acids to flavor-associated compounds is controversial. The objective of this study was to determine the ability of Brevibacterium linens BL2 to produce fatty acids from amino acids and alpha-keto acids and determine the occurrence of the likely genes in the draft genome sequence. BL2 catabolized amino acids to fatty acids only under carbohydrate starvation conditions. The primary fatty acid end products from leucine were isovaleric acid, acetic acid, and propionic acid. In contrast, logarithmic-phase cells of BL2 produced fatty acids from alpha-keto acids only. BL2 also converted alpha-keto acids to branched-chain fatty acids after carbohydrate starvation was achieved. At least 100 genes are potentially involved in five different metabolic pathways. The genome of B. linens ATCC 9174 contained these genes for production and degradation of fatty acids. These data indicate that brevibacteria have the ability to produce fatty acids from amino and alpha-keto acids and that carbon metabolism is important in regulating this event. PMID- 15528497 TI - Air- and dustborne mycoflora in houses free of water damage and fungal growth. AB - Typically, studies on indoor fungal growth in buildings focus on structures with known or suspected water damage, moisture, and/or indoor fungal growth problems. Reference information on types of culturable fungi and total fungal levels are generally not available for buildings without these problems. This study assessed 50 detached single-family homes in metropolitan Atlanta, Ga., to establish a baseline of "normal and typical" types and concentrations of airborne and dustborne fungi in urban homes which were predetermined not to have noteworthy moisture problems or indoor fungal growth. Each home was visually examined, and samples of indoor and outdoor air and of indoor settled dust were taken in winter and summer. The results showed that rankings by prevalence and abundance of the types of airborne and dustborne fungi did not differ from winter to summer, nor did these rankings differ when air samples taken indoors were compared with those taken outdoors. Water indicator fungi were essentially absent from both air and dust samples. The air and dust data sets were also examined specifically for the proportions of colonies from ecological groupings such as leaf surface fungi and soil fungi. In the analysis of dust for culturable fungal colonies, leaf surface fungi constituted a considerable portion (>20%) of the total colonies in at least 85% of the samples. Thus, replicate dust samples with less than 20% of colonies from leaf surface fungi are unlikely to be from buildings free of moisture or mold growth problems. PMID- 15528498 TI - Serologic and molecular characterization of Vibrio parahaemolyticus strains isolated from seawater and fish products of the Gulf of Mexico. AB - The thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) are the main virulence factors of Vibrio parahaemolyticus. We isolated V. parahaemolyticus from seawater, fish, and oysters obtained from the Pueblo Viejo Lagoon in Veracruz, determined the serogroups, phenotypically and genotypically characterized TDH and TRH, and investigated the presence of the toxR gene. A total of 46 V. parahaemolyticus strains were isolated, and all of them amplified the 368-bp toxR gene fragment. The trh gene was not identified in any of the strains; 4 of the 46 strains were Kanagawa phenomenon (KP) positive and amplified the 251-bp tdh gene fragment. The most frequent serogroup was serogroup O3. This is the first report of the presence of KP-positive tdh-positive environmental V. parahaemolyticus strains in Mexico. PMID- 15528499 TI - Effect of microbial inoculants on the indigenous actinobacterial endophyte population in the roots of wheat as determined by terminal restriction fragment length polymorphism. AB - The effect of single actinobacterial endophyte seed inoculants and a mixed microbial soil inoculant on the indigenous endophytic actinobacterial population in wheat roots was investigated by using the molecular technique terminal restriction fragment length polymorphism (T-RFLP). Wheat was cultivated either from seeds coated with the spores of single pure actinobacterial endophytes of Microbispora sp. strain EN2, Streptomyces sp. strain EN27, and Nocardioides albus EN46 or from untreated seeds sown in soil with and without a commercial mixed microbial soil inoculant. The endophytic actinobacterial population within the roots of 6-week-old wheat plants was assessed by T-RFLP. Colonization of the wheat roots by the inoculated actinobacterial endophytes was detected by T-RFLP, as were 28 to 42 indigenous actinobacterial genera present in the inoculated and uninoculated plants. The presence of the commercial mixed inoculant in the soil reduced the endophytic actinobacterial diversity from 40 genera to 21 genera and reduced the detectable root colonization by approximately half. The results indicate that the addition of a nonadapted microbial inoculum to the soil disrupted the natural actinobacterial endophyte population, reducing diversity and colonization levels. This was in contrast to the addition of a single actinobacterial endophyte to the wheat plant, where the increase in colonization level could be confirmed even though the indigenous endophyte population was not adversely affected. PMID- 15528500 TI - Relationships of a novel Lyme disease spirochete, Borrelia spielmani sp. nov., with its hosts in Central Europe. AB - To determine whether the pathogenic variant of Lyme disease spirochetes, isolate A14S, is perpetuated in a particular reservoir-vector relationship, we screened vector ticks in various Central European sites for a related spirochete and determined its host association. A14S-like spirochetes infect numerous questing ticks in the Petite Camargue Alsacienne (PC). They frequently infect dormice, but no mice or voles. Garden dormice appear to be better reservoir hosts for A14S like spirochetes than for Borrelia afzelii, because these spirochetes are retained longer and infect ticks more readily. Spirochetes associated with garden dormice in the PC site form a homologous entity with those isolated from a human patient in The Netherlands. Its unique biological relationship together with previous genetic characterization justifies designating this dormouse-associated genospecies as a distinct entity. Garden dormice serve as the main reservoir hosts of a novel genospecies, Borrelia spielmani sp. nov., one of several that cause Lyme disease in people. PMID- 15528501 TI - Escherichia coli contamination of vegetables grown in soils fertilized with noncomposted bovine manure: garden-scale studies. AB - In this study we tested the validity of the National Organic Program (NOP) requirement for a > or =120-day interval between application of noncomposted manure and harvesting of vegetables grown in manure-fertilized soil. Noncomposted bovine manure was applied to 9.3-m2 plots at three Wisconsin sites (loamy sand, silt loam, and silty clay loam) prior to spring and summer planting of carrots, radishes, and lettuce. Soil and washed (30 s under running tap water) vegetables were analyzed for indigenous Escherichia coli. Within 90 days, the level of E. coli in manure-fertilized soil generally decreased by about 3 log CFU/g from initial levels of 4.2 to 4.4 log CFU/g. Low levels of E. coli generally persisted in manure-fertilized soil for more than 100 days and were detected in enriched soil from all three sites 132 to 168 days after manure application. For carrots and lettuce, at least one enrichment-negative sample was obtained < or =100 days after manure application for 63 and 88% of the treatments, respectively. The current > or =120-day limit provided an even greater likelihood of not detecting E. coli on carrots (> or =1 enrichment-negative result for 100% of the treatments). The rapid maturation of radishes prevented conclusive evaluation of a 100- or 120-day application-to-harvest interval. The absolute absence of E. coli from vegetables harvested from manure-fertilized Wisconsin soils may not be ensured solely by adherence to the NOP > or =120-day limit. Unless pathogens are far better at colonizing vegetables than indigenous E. coli strains are, it appears that the risk of contamination for vegetables grown in Wisconsin soils would be elevated only slightly by reducing the NOP requirement to > or =100 days. PMID- 15528502 TI - Redox transformations of arsenic oxyanions in periphyton communities. AB - Periphyton (Cladophora sp.) samples from a suburban stream lacking detectable dissolved As were able to reduce added As(V) to As(III) when incubated under anoxic conditions and, conversely, oxidized added As(III) to As(V) with aerobic incubation. Both types of activity were abolished in autoclaved controls, thereby demonstrating its biological nature. The reduction of As(V) was inhibited by chloramphenicol, indicating that it required the synthesis of new protein. Nitrate also inhibited As(V) reduction, primarily because it served as a preferred electron acceptor to which the periphyton community was already adapted. However, part of the inhibition was also caused by microbial reoxidation of As(III) linked to nitrate. Addition of [14C]glucose to anoxic samples resulted in the production of 14CO2, suggesting that the observed As(V) reduction was a respiratory process coupled to the oxidation of organic matter. The population density of As(V)-reducing bacteria within the periphyton increased with time and with the amount of As(V) added, reaching values as high as approximately 10(6) cells ml(-1) at the end of the incubation. This indicated that dissimilatory As(V) reduction in these populations was linked to growth. However, As(V) respiring bacteria were found to be present, albeit at lower numbers (approximately 10(2) ml(-1)), in freshly sampled periphyton. These results demonstrate the presence of a bacterial population within the periphyton communities that is capable of two key arsenic redox transformations that were previously studied in As-contaminated environments, which suggests that these processes are widely distributed in nature. This assumption was reinforced by experiments with estuarine samples of Cladophora sericea in which we detected a similar capacity for anaerobic As(V) reduction and aerobic As(III) oxidation. PMID- 15528503 TI - Survival of Shewanella oneidensis MR-1 after UV radiation exposure. AB - We systematically investigated the physiological response as well as DNA damage repair and damage tolerance in Shewanella oneidensis MR-1 following UVC, UVB, UVA, and solar light exposure. MR-1 showed the highest UVC sensitivity among Shewanella strains examined, with D37 and D10 values of 5.6 and 16.5% of Escherichia coli K-12 values. Stationary cells did not show an increased UVA resistance compared to exponential-phase cells; instead, they were more sensitive at high UVA dose. UVA-irradiated MR-1 survived better on tryptic soy agar than Luria-Bertani plates regardless of the growth stage. A 20% survival rate of MR-1 was observed following doses of 3.3 J of UVC m(-2), 568 J of UVB m(-2), 25 kJ of UVA m(-2), and 558 J of solar UVB m(-2), respectively. Photoreactivation conferred an increased survival rate to MR-1 of as much as 177- to 365-fold, 11- to 23-fold, and 3- to 10-fold following UVC, UVB, and solar light irradiation, respectively. A significant UV mutability to rifampin resistance was detected in both UVC- and UVB-treated samples, with the mutation frequency in the range of 10(-5) to 10(-6). Unlike in E. coli, the expression levels of the nucleotide excision repair (NER) component genes uvrA, uvrB, and uvrD were not damage inducible in MR-1. Complementation of Pseudomonas aeruginosa UA11079 (uvrA deficient) with uvrA of MR-1 increased the UVC survival of this strain by more than 3 orders of magnitude. Loss of damage inducibility of the NER system appears to contribute to the high sensitivity of this bacterium to UVR as well as to other DNA-damaging agents. PMID- 15528504 TI - Diversity and distribution of Frankia strains symbiotic with Ceanothus in California. AB - Frankia strains symbiotic with Ceanothus present an interesting opportunity to study the patterns and causes of Frankia diversity and distribution within a particular host infectivity group. We intensively sampled Frankia from nodules on Ceanothus plants along an elevational gradient in the southern Sierra Nevada of California, and we also collected nodules from a wider host taxonomic and geographic range throughout California. The two sampling scales comprised 36 samples from eight species of Ceanothus representing six of the seven major biogeographic regions in and around California. The primary objective of this study was to use a quantitative model to test the relative importance of geographic separation, host specificity, and environment in influencing the identity of Ceanothus Frankia symbionts as determined by ribosomal DNA sequence data. At both sampling scales, Frankia strains symbiotic with Ceanothus exhibited a high degree of genetic similarity. Frankia strains symbiotic with Chamaebatia (Rosaceae) were within the same clade as several Ceanothus symbionts. Results from a classification and regression tree model used to quantitatively explain Frankia phylogenetic groupings demonstrated that the only significant variable in distinguishing between phylogenetic groups at the more local sampling scale was host species. At the regional scale, Frankia phylogenetic groupings were explained by host species and the biogeographic province of sample collection. We did not find any significant correspondence between Frankia and Ceanothus phylogenies indicative of coevolution, but we concluded that the identity of Frankia strains inhabiting Ceanothus nodules may involve interactions between host species specificity and geographic isolation. PMID- 15528505 TI - Use of merocyanine 540 for photodynamic inactivation of Staphylococcus aureus planktonic and biofilm cells. AB - Photodynamic inactivation of Staphylococcus aureus planktonic and biofilm cells by a phtotosensitizer, merocyanine 540 (MC 540), was investigated. For the planktonic experiments, MC 540 binding efficiency to bacterial cells was found to increase with both increasing MC 540 concentration and increasing incubation time, but the binding became saturated following 10 min of incubation. The antimicrobial activity was enhanced with an increasing light dose, but an increase in the light dose could not further improve the antimicrobial activity if the maximum excitation level attainable was less than the necessary minimum threshold level. Complete inactivation was achieved when the excitation level of MC 540 was somewhere above the threshold level. The relationship between antimicrobial activity and the excitation level of MC 540 revealed that the more MC 540 was excited, the more S. aureus cells were killed. For the biofilm experiments, the antimicrobial activity was enhanced with an increase in the light dose. No viable cells were detected when organisms were exposed to 15 mug of MC 540 per ml and a light dose of 600 J/cm2 or to 20 mug of MC 540 per ml and a light dose of 450 J/cm2. A quantitative analysis of MC 540 bound to biofilms was also performed, and the images from confocal laser scanning microscopy provided direct evidence that revealed the difference between the MC 540 remaining in the biofilms prior to irradiation and the MC 540 remaining in the biofilms after irradiation. The results of both the planktonic and biofilm experiments suggest that the antimicrobial activity of photodynamic inactivation of S. aureus is closely related to the excitation level of MC 540. PMID- 15528506 TI - Real-time PCR quantitation of clostridia in feces of autistic children. AB - Based on the hypothesis that intestinal clostridia play a role in late-onset autism, we have been characterizing clostridia from stools of autistic and control children. We applied the TaqMan real-time PCR procedure to detect and quantitate three Clostridium clusters and one Clostridium species, C. bolteae, in stool specimens. Group- and species-specific primers targeting the 16S rRNA genes were designed, and specificity of the primers was confirmed with DNA from related bacterial strains. In this procedure, a linear relationship exists between the threshold cycle (CT) fluorescence value and the number of bacterial cells (CFU). The assay showed high sensitivity: as few as 2 cells of members of cluster I, 6 cells of cluster XI, 4 cells of cluster XIVab, and 0.6 cell of C. bolteae could be detected per PCR. Analysis of the real-time PCR data indicated that the cell count differences between autistic and control children for C. bolteae and the following Clostridium groups were statistically significant: mean counts of C. bolteae and clusters I and XI in autistic children were 46-fold (P = 0.01), 9.0 fold (P = 0.014), and 3.5-fold (P = 0.004) greater than those in control children, respectively, but not for cluster XIVab (2.6 x 10(8) CFU/g in autistic children and 4.8 x 10(8) CFU/g in controls; respectively). More subjects need to be studied. The assay is a rapid and reliable method, and it should have great potential for quantitation of other bacteria in the intestinal tract. PMID- 15528507 TI - Measurements of fitness and competition in commensal Escherichia coli and E. coli O157:H7 strains. AB - Although the main reservoirs for pathogenic Escherichia coli O157:H7 are cattle and the cattle environment, factors that affect its tenure in the bovine host and its survival outside humans and cattle have not been well studied. It is also not understood what physiological properties, if any, distinguish these pathogens from commensal counterparts that live as normal members of the human and bovine gastrointestinal tracts. To address these questions, individual and competitive fitness experiments, indirect antagonism assays, and antibiotic resistance and carbon utilization analyses were conducted using a strain set consisting of 122 commensal and pathogenic strains. The individual fitness experiments, under four different environments (rich medium, aerobic and anaerobic; rumen medium, anaerobic; and a minimal medium, aerobic) revealed no differences in growth rates between commensal E. coli and E. coli O157:H7 strains. Indirect antagonism assays revealed that E. coli O157:H7 strains more frequently produced inhibitory substances than commensal strains did, under the conditions tested, although both groups displayed moderate sensitivity. Only minor differences were noted in the antibiotic resistance patterns of the two groups. In contrast, several differences between commensal and O157:H7 groups were observed based on their carbon utilization profiles. Of 95 carbon sources tested, 27 were oxidized by commensal E. coli strains but not by the E. coli O157:H7 strains. Despite the observed physiological and biochemical differences between these two groups of E. coli strains, however, the O157:H7 strains did not appear to possess traits that would confer advantages in the bovine or extraintestinal environment. PMID- 15528508 TI - Stages of infection during the tripartite interaction between Xenorhabdus nematophila, its nematode vector, and insect hosts. AB - Bacteria of the genus Xenorhabdus are mutually associated with entomopathogenic nematodes of the genus Steinernema and are pathogenic to a broad spectrum of insects. The nematodes act as vectors, transmitting the bacteria to insect larvae, which die within a few days of infection. We characterized the early stages of bacterial infection in the insects by constructing a constitutive green fluorescent protein (GFP)-labeled Xenorhabdus nematophila strain. We injected the GFP-labeled bacteria into insects and monitored infection. We found that the bacteria had an extracellular life cycle in the hemolymph and rapidly colonized the anterior midgut region in Spodoptera littoralis larvae. Electron microscopy showed that the bacteria occupied the extracellular matrix of connective tissues within the muscle layers of the Spodoptera midgut. We confirmed the existence of such a specific infection site in the natural route of infection by infesting Spodoptera littoralis larvae with nematodes harboring GFP-labeled Xenorhabdus. When the infective juvenile (IJ) nematodes reached the insect gut, the bacterial cells were rapidly released from the intestinal vesicle into the nematode intestine. Xenorhabdus began to escape from the anus of the nematodes when IJs were wedged in the insect intestinal wall toward the insect hemolymph. Following their release into the insect hemocoel, GFP-labeled bacteria were found only in the anterior midgut region and hemolymph of Spodoptera larvae. Comparative infection assays conducted with another insect, Locusta migratoria, also showed early bacterial colonization of connective tissues. This work shows that the extracellular matrix acts as a particular colonization site for X. nematophila within insects. PMID- 15528509 TI - High-rate nitrification at low pH in suspended- and attached-biomass reactors. AB - This article reports on high-rate nitrification at low pH in biofilm and suspended-biomass reactors by known chemolithotrophic bacteria. In the biofilm reactor, at low pH (4.3 +/- 0.1) and low bulk ammonium concentrations (9.3 +/- 3.3 mg.liter(-1)), a very high nitrification rate of 5.6 g of N oxidized.liter( 1).day(-1) was achieved. The specific nitrification rate (0.55 g of N.g of biomass(-1).day(-1)) was similar to values reported for nitrifying reactors at optimal pH. In the suspended-biomass reactor, the average pH was significantly lower than that in the biofilm reactor (pH 3.8 +/- 0.3), and values as low as pH 3.2 were found. In addition, measurements in the suspended-biomass reactor, using isotope-labeled ammonium (15N), showed that in spite of the very low pH, biomass growth occurred with a yield of 0.1 g of biomass.g of N oxidized(-1). Fluorescence in situ hybridization using existing rRNA-targeted oligonucleotide probes showed that the nitrifying bacteria were from the monophyletic genus Nitrosomonas, suggesting that autotrophic nitrification at low pH is more widespread than previously thought. The results presented in this paper clearly show that autotrophic nitrifying bacteria have the ability to nitrify at a high rate at low pH and in the presence of only a negligible free ammonia concentration, suggesting the presence of an efficient ammonium uptake system and the means to cope with low pH. PMID- 15528510 TI - Identification of a third msa gene in Renibacterium salmoninarum and the associated virulence phenotype. AB - Renibacterium salmoninarum, a gram-positive diplococcobacillus, causes bacterial kidney disease, a condition that can result in extensive morbidity and mortality among stocks of fish. An immunodominant extracellular protein, called major soluble antigen (MSA), is encoded by two identical genes, msa1 and msa2. We found evidence for a third msa gene, msa3, which appears to be a duplication of msa1. Unlike msa1 and msa2, msa3 is not present in all isolates of R. salmoninarum. The presence of the msa3 locus does not affect total MSA production in culture conditions. In a challenge study, isolates possessing the msa3 locus reduced median survival in juvenile chinook salmon (Oncorhynchus tshawytscha) by an average of 34% at doses of < or =10(5) cells per fish compared to isolates lacking the msa3 locus. In contrast, no difference in survival was observed at the highest dose, 10(6) cells per fish. The phenotype associated with the msa3 locus and its nonuniform distribution may contribute to observed differences in virulence among R. salmoninarum isolates. PMID- 15528511 TI - Transport of the harmful bloom alga Aureococcus anophagefferens by oceangoing ships and coastal boats. AB - It is well established that cyst-forming phytoplankton species are transported in ships' ballast tanks. However, there is increasing evidence that other phytoplankton species which do not encyst are also capable of surviving ballast transit. These species have alternative modes of nutrition (hetero- or mixotrophy) and/or are able to survive long-term darkness. In our studies of no ballast-on-board vessels arriving in the Great Lakes, we tested for the presence of the harmful algal bloom species Aureococcus anophagefferens (brown tide) in residual (i.e., unpumpable) ballast water using methods based on the PCR. During 2001, the brown tide organism was detected in 7 of 18 ballast water tanks in commercial ships following transit from foreign ports. Furthermore, it was detected after 10 days of ballast tank confinement during a vessel transit in the Great Lakes, a significant result given the large disparity between the salinity tolerance for active growth of Aureococcus (>22 ppt) and the low salinity of the residual ballast water (approximately 2 ppt). We also investigated the potential for smaller, recreational vessels to transport and distribute Aureococcus. During the summer of 2002, 11 trailered boats from the inland bays of Delaware and coastal bays of Maryland were sampled. Brown tide was detected in the bilge water in the bottoms of eight boats, as well as in one live-well sample. Commercial ships and small recreational boats are therefore implicated as potential vectors for long-distance transport and local-scale dispersal of Aureococcus. PMID- 15528512 TI - Frequency and spatial distribution of environmental Campylobacter spp. AB - Humans are exposed to Campylobacter spp. in a range of sources via both food and environmental pathways. For this study, we explored the frequency and distribution of thermophilic Campylobacter spp. in a 10- by 10-km square rural area of Cheshire, United Kingdom. The area contains approximately 70, mainly dairy, farms and is used extensively for outdoor recreational activities. Campylobacter spp. were isolated from a range of environmental samples by use of a systematic sampling grid. Livestock (mainly cattle) and wildlife feces and environmental water and soil samples were cultured, and isolates were presumptively identified by standard techniques. These isolates were further characterized by PCR. Campylobacter jejuni was the most prevalent species in all animal samples, ranging from 11% in samples from nonavian wildlife to 36% in cattle feces, and was isolated from 15% of water samples. Campylobacter coli was commonly found in water (17%) and sheep (21%) samples, but rarely in other samples. Campylobacter lari was recovered from all sample types, with the exception of sheep feces, and was found in moderate numbers in birds (7%) and water (5%). Campylobacter hyointestinalis was only recovered from cattle (7%) and birds (1%). The spatial distribution and determinants of C. jejuni in cattle feces were examined by the use of model-based spatial statistics. The distribution was consistent with very localized within-farm or within-field transmission and showed little evidence of any larger-scale spatial dependence. We concluded that there is a potentially high risk of human exposure to Campylobacter spp., particularly C. jejuni, in the environment of our study area. The prevalence and likely risk posed by C. jejuni-positive cattle feces in the environment diminished as the fecal material aged. After we took into account the age of the fecal material, the absence or presence of rain, and the presence of bird feces, there was evidence of significant variation in the prevalence of C. jejuni-positive cattle feces between grazing fields but no evidence of spatial clustering beyond this resolution. The spatial pattern of C. jejuni is therefore consistent with that for an organism that is ubiquitous in areas contaminated with cattle feces, with a short-scale variation in infection intensity that cannot be explained solely by variations in the age of the fecal material. The observed pattern is not consistent with large-scale transmission attributable to watercourses, wildlife territories, or other geographical features that transcend field and farm boundaries. PMID- 15528513 TI - No significant contribution of arbuscular mycorrhizal fungi to transfer of radiocesium from soil to plants. AB - The diffuse pollution by fission and activation products following nuclear accidents and weapons testing is of major public concern. Among the nuclides that pose a serious risk if they enter the human food chain are the cesium isotopes 137Cs and 134Cs (with half-lives of 30 and 2 years, respectively). The biogeochemical cycling of these isotopes in forest ecosystems is strongly affected by their preferential absorption in a range of ectomycorrhiza-forming basidiomycetes. An even more widely distributed group of symbiotic fungi are the arbuscular mycorrhizal fungi, which colonize most herbaceous plants, including many agricultural crops. These fungi are known to be more efficient than ectomycorrhizas in transporting mineral elements from soil to plants. Their role in the biogeochemical cycling of Cs is poorly known, in spite of the consequences that fungal Cs transport may have for transfer of Cs into the human food chain. This report presents the first data on transport of Cs by these fungi by use of radiotracers and compartmented growth systems where uptake by roots and mycorrhizal hyphae is distinguished. Independent experiments in three laboratories that used different combinations of fungi and host plants all demonstrated that these fungi do not contribute significantly to plant uptake of Cs. The implications of these findings for the bioavailability of radiocesium in different terrestrial ecosystems are discussed. PMID- 15528514 TI - Aflatoxin biosynthesis cluster gene cypA is required for G aflatoxin formation. AB - Aspergillus flavus isolates produce only aflatoxins B1 and B2, while Aspergillus parasiticus and Aspergillus nomius produce aflatoxins B1, B2, G1, and G2. Sequence comparison of the aflatoxin biosynthesis pathway gene cluster upstream from the polyketide synthase gene, pksA, revealed that A. flavus isolates are missing portions of genes (cypA and norB) predicted to encode, respectively, a cytochrome P450 monooxygenase and an aryl alcohol dehydrogenase. Insertional disruption of cypA in A. parasiticus yielded transformants that lack the ability to produce G aflatoxins but not B aflatoxins. The enzyme encoded by cypA has highest amino acid identity to Gibberella zeae Tri4 (38%), a P450 monooxygenase previously shown to be involved in trichodiene epoxidation. The substrate for CypA may be an intermediate formed by oxidative cleavage of the A ring of O methylsterigmatocystin by OrdA, the P450 monooxygenase required for formation of aflatoxins B1 and B2. PMID- 15528515 TI - Coupling of functional gene diversity and geochemical data from environmental samples. AB - Genomic techniques commonly used for assessing distributions of microorganisms in the environment often produce small sample sizes. We investigated artificial neural networks for analyzing the distributions of nitrite reductase genes (nirS and nirK) and two sets of dissimilatory sulfite reductase genes (dsrAB1 and dsrAB2) in small sample sets. Data reduction (to reduce the number of input parameters), cross-validation (to measure the generalization error), weight decay (to adjust model parameters to reduce generalization error), and importance analysis (to determine which variables had the most influence) were useful in developing and interpreting neural network models that could be used to infer relationships between geochemistry and gene distributions. A robust relationship was observed between geochemistry and the frequencies of genes that were not closely related to known dissimilatory sulfite reductase genes (dsrAB2). Uranium and sulfate appeared to be the most related to distribution of two groups of these unusual dsrAB-related genes. For the other three groups, the distributions appeared to be related to pH, nickel, nonpurgeable organic carbon, and total organic carbon. The models relating the geochemical parameters to the distributions of the nirS, nirK, and dsrAB1 genes did not generalize as well as the models for dsrAB2. The data also illustrate the danger (generating a model that has a high generalization error) of not using a validation approach in evaluating the meaningfulness of the fit of linear or nonlinear models to such small sample sizes. PMID- 15528516 TI - Glutamine, glutamate, and alpha-glucosylglycerate are the major osmotic solutes accumulated by Erwinia chrysanthemi strain 3937. AB - Erwinia chrysanthemi is a phytopathogenic soil enterobacterium closely related to Escherichia coli. Both species respond to hyperosmotic pressure and to external added osmoprotectants in a similar way. Unexpectedly, the pools of endogenous osmolytes show different compositions. Instead of the commonly accumulated glutamate and trehalose, E. chrysanthemi strain 3937 promotes the accumulation of glutamine and alpha-glucosylglycerate, which is a new osmolyte for enterobacteria, together with glutamine. The amounts of the three osmolytes increased with medium osmolarity and were reduced when betaine was provided in the growth medium. Both glutamine and glutamate showed a high rate of turnover, whereas glucosylglycerate stayed stable. In addition, the balance between the osmolytes depended on the osmolality of the medium. Glucosylglycerate and glutamate were the major intracellular compounds in low salt concentrations, whereas glutamine predominated at higher concentrations. Interestingly, the ammonium content of the medium also influenced the pool of osmolytes. During bacterial growth with 1 mM ammonium in stressing conditions, more glucosylglycerate accumulated by far than the other organic solutes. Glucosylglycerate synthesis has been described in some halophilic archaea and bacteria but not as a dominant osmolyte, and its role as an osmolyte in Erwinia chrysanthemi 3937 shows that nonhalophilic bacteria can also use ionic osmolytes. PMID- 15528517 TI - Representative freshwater bacterioplankton isolated from Crater Lake, Oregon. AB - High-throughput culturing (HTC) methods that rely on dilution to extinction in very-low-nutrient media were used to obtain bacterial isolates from Crater Lake, Oregon. 16S rRNA sequence determination and phylogenetic reconstruction were used to determine the potential ecological significance of isolated bacteria, both in Crater Lake and globally. Fifty-five Crater Lake isolates yielded 16 different 16S rRNA gene sequences. Thirty of 55 (55%) Crater Lake isolates had 16S rRNA gene sequences with 97% or greater similarity to sequences recovered previously from Crater Lake 16S rRNA gene clone libraries. Furthermore, 36 of 55 (65%) Crater Lake isolates were found to be members of widely distributed freshwater groups. These results confirm that HTC is a significant improvement over traditional isolation techniques that tend to enrich for microorganisms that do not predominate in their environment and rarely correlate with 16S rRNA gene clone library sequences. Although all isolates were obtained under dark, heterotrophic growth conditions, 2 of the 16 different groups showed evidence of photosynthetic capability as assessed by the presence of puf operon sequences, suggesting that photoheterotrophy may be a significant process in this oligotrophic, freshwater habitat. PMID- 15528518 TI - Bacterium-based NO2- biosensor for environmental applications. AB - A sensitive NO2- biosensor that is based on bacterial reduction of NO2- to N2O and subsequent detection of the N2O by a built-in electrochemical N2O sensor was developed. Four different denitrifying organisms lacking NO3- reductase activity were assessed for use in the biosensor. The relevant physiological aspects examined included denitrifying characteristics, growth rate, NO2- tolerance, and temperature and salinity effects on the growth rate. Two organisms were successfully used in the biosensor. The preferred organism was Stenotrophomonas nitritireducens, which is an organism with a denitrifying pathway deficient in both NO3- and N2O reductases. Alternatively Alcaligenes faecalis could be used when acetylene was added to inhibit its N2O reductase. The macroscale biosensors constructed exhibited a linear NO2- response at concentrations up to 1 to 2 mM. The detection limit was around 1 microM NO2-, and the 90% response time was 0.5 to 3 min. The sensor signal was specific for NO2-, and interference was observed only with NH2OH, NO, N2O, and H2S. The sensor signal was affected by changes in temperature and salinity, and calibration had to be performed in a system with a temperature and an ionic strength comparable to those of the medium analyzed. A broad range of water bodies could be analyzed with the biosensor, including freshwater systems, marine systems, and oxic-anoxic wastewaters. The NO2- biosensor was successfully used for long-term online monitoring in wastewater. Microscale versions of the NO2- biosensor were constructed and used to measure NO2- profiles in marine sediment. PMID- 15528519 TI - Phylogenetic characterization of methanogenic assemblages in eutrophic and oligotrophic areas of the Florida Everglades. AB - Agricultural activities have produced well-documented changes in the Florida Everglades, including establishment of a gradient in phosphorus concentrations in Water Conservation Area 2A (WCA-2A) of the northern Everglades. An effect of increased phosphorus concentrations is increased methanogenesis in the eutrophic regions compared to the oligotrophic regions of WCA-2A. The goal of this study was to identify relationships between eutrophication and composition and activity of methanogenic assemblages in WCA-2A soils. Distributions of two genes associated with methanogens were characterized in soils taken from WCA-2A: the archaeal 16S rRNA gene and the methyl coenzyme M reductase gene. The richness of methanogen phylotypes was greater in eutrophic than in oligotrophic sites, and sequences related to previously cultivated and uncultivated methanogens were found. A preferential selection for the order Methanomicrobiales was observed in mcrA clone libraries, suggesting primer bias for this group. A greater diversity within the Methanomicrobiales was observed in mcrA clone libraries than in 16S rRNA gene libraries. 16S rRNA phylogenetic analyses revealed a dominance of clones related to Methanosaeta spp., an acetoclastic methanogen dominant in environments with low acetate concentrations. A significant number of clones were related to Methanomicrobiales, an order characterized by species utilizing hydrogen and formate as methanogenic substrates. No representatives of the orders Methanobacteriales and Methanococcales were found in any 16S rRNA clone library, although some Methanobacteriales were found in mcrA libraries. Hydrogenotrophs are the dominant methanogens in WCA-2A, and acetoclastic methanogen genotypes that proliferate in low acetate concentrations outnumber those that typically dominate in higher acetate concentrations. PMID- 15528520 TI - Diversity and antagonistic potential of bacteria associated with bryophytes from nutrient-poor habitats of the Baltic Sea Coast. AB - Very little is known about the interaction of bryophytes with bacteria. Therefore, we analyzed bacteria associated with three bryophyte species, Tortula ruralis, Aulacomnium palustre, and Sphagnum rubellum, which represent typical moss species of three nutrient-poor plant communities at the southern Baltic Sea coast in Germany. By use of two cultivation-independent techniques, denaturing gradient gel electrophoresis and single-strand conformation polymorphism analysis of the 16S ribosomal DNA, a high degree of moss specificity was found for associated bacterial communities. This specificity could be further evidenced by a cultivation-dependent approach for the following parameters: (i) plate counts of bacteria on R2A medium, (ii) proportion of antagonistic isolates, (iii) antagonistic activity as well as spectrum against pathogens, and (iv) diversity and richness of antagonistic isolates. The proportion of isolates with antagonistic activity against the pathogenic model fungus Verticillium dahliae was highest for S. rubellum (31%), followed by A. palustre (17%) and T. ruralis (5%). A high percentage (99%) of moss-associated antagonistic bacteria produced antifungal compounds. The high recovery of antagonistic isolates strongly suggests that bryophytes represent an ecological niche which harbors a diverse and hitherto largely uncharacterized microbial population with yet unknown and untapped potential biotechnological applications, e.g., for biological control of plant pathogens. PMID- 15528521 TI - Novel endophytic nitrogen-fixing clostridia from the grass Miscanthus sinensis as revealed by terminal restriction fragment length polymorphism analysis. AB - Anaerobic nitrogen-fixing consortia consisting of N2-fixing clostridia and diverse nondiazotrophic bacteria were previously isolated from various gramineous plants (K. Minamisawa, K. Nishioka, T. Miyaki, B. Ye, T. Miyamoto, M. You, A. Saito, M. Saito, W. Barraquio, N. Teaumroong, T. Sein, and T. Tadashi, Appl. Environ. Microbiol. 70:3096-3102, 2004). For this work, clostridial populations and their phylogenetic structures in a stand of the grass Miscanthus sinensis in Japan were assessed by a 16S rRNA gene-targeted terminal restriction fragment length polymorphism (TRFLP) analysis combined with most-probable-number (MPN) counts. PCR primers and restriction enzymes were optimized for analyses of the plant clostridia. Clostridia were detected in strongly surface-sterilized leaves, stems, and roots of the plants at approximately 10(4) to 10(5) cells/g of fresh weight; they made up a large proportion of N2-fixing bacterial populations, as determined by MPN counts associated with an acetylene reduction assay. Phylogenetic grouping by MPN-TRFLP analysis revealed that the clostridial populations belonged to group II of cluster XIVa and groups IV and V of cluster I; this result was supported by a culture-independent TRFLP analysis using direct DNA extraction from plants. When phylogenetic populations from M. sinensis and the soil around the plants were compared, group II clostridia were found to exist exclusively in M. sinensis. PMID- 15528522 TI - Display of biologically functional insecticidal toxin on the surface of lambda phage. AB - The successful use of Bacillus thuringiensis insecticidal toxins to control agricultural pests could be undermined by the evolution of insect resistance. Under selection pressure in the laboratory, a number of insects have gained resistance to the toxins, and several cases of resistance in the diamondback moth have been reported from the field. The use of protein engineering to develop novel toxins active against resistant insects could offer a solution to this problem. The display of proteins on the surface of phages has been shown to be a powerful technology to search for proteins with new characteristics from combinatorial libraries. However, this potential of phage display to develop Cry toxins with new binding properties and new target specificities has hitherto not been realized because of the failure of displayed Cry toxins to bind their natural receptors. In this work we describe the construction of a display system in which the Cry1Ac toxin is fused to the amino terminus of the capsid protein D of bacteriophage lambda. The resultant phage was viable and infectious, and the displayed toxin interacted successfully with its natural receptor. PMID- 15528523 TI - Effects of the metalloid oxyanion tellurite (TeO32-) on growth characteristics of the phototrophic bacterium Rhodobacter capsulatus. AB - This work examines the effects of potassium tellurite (K2TeO3) on the cell viability of the facultative phototroph Rhodobacter capsulatus. There was a growth mode-dependent response in which cultures anaerobically grown in the light tolerate the presence of up to 250 to 300 microg of tellurite (TeO3(2-)) per ml, while dark-grown aerobic cells were inhibited at tellurite levels as low as 2 microg/ml. The tellurite sensitivity of aerobic cultures was evident only for growth on minimal salt medium, whereas it was not seen during growth on complex medium. Notably, through the use of flow cytometry, we show that the cell membrane integrity was strongly affected by tellurite during the early growth phase (< or =50% viable cells); however, at the end of the growth period and in parallel with massive tellurite intracellular accumulation as elemental Te0 crystallites, recovery of cytoplasmic membrane integrity was apparent (> or =90% viable cells), which was supported by the development of a significant membrane potential (Deltapsi = 120 mV). These data are taken as evidence that in anaerobic aquatic habitats, the facultative phototroph R. capsulatus might act as a natural scavenger of the highly soluble and toxic oxyanion tellurite. PMID- 15528524 TI - Multiplex nucleic acid sequence-based amplification for simultaneous detection of several enteric viruses in model ready-to-eat foods. AB - Human enteric viruses are currently recognized as one of the most important causes of food-borne disease. Implication of enteric viruses in food-borne outbreaks can be difficult to confirm due to the inadequacy of the detection methods available. In this study, a nucleic acid sequence-based amplification (NASBA) method was developed in a multiplex format for the specific, simultaneous, and rapid detection of epidemiologically relevant human enteric viruses. Three previously reported primer sets were used in a single reaction for the amplification of RNA target fragments of 474, 371, and 165 nucleotides for the detection of hepatitis A virus and genogroup I and genogroup II noroviruses, respectively. Amplicons were detected by agarose gel electrophoresis and confirmed by electrochemiluminescence and Northern hybridization. Endpoint detection sensitivity for the multiplex NASBA assay was approximately 10(-1) reverse transcription-PCR-detectable units (or PFU, as appropriate) per reaction. When representative ready-to-eat foods (deli sliced turkey and lettuce) were inoculated with various concentrations of each virus and processed for virus detection with the multiplex NASBA method, all three human enteric viruses were simultaneously detected at initial inoculum levels of 10(0) to 10(2) reverse transcription-PCR-detectable units (or PFU)/9 cm2 in both food commodities. The multiplex NASBA system provides rapid and simultaneous detection of clinically relevant food-borne viruses in a single reaction tube and may be a promising alternative to reverse transcription-PCR for the detection of viral contamination of foods. PMID- 15528525 TI - Real-time PCR for detection and quantification of the protistan parasite Perkinsus marinus in environmental waters. AB - The protistan parasite Perkinsus marinus is a severe pathogen of the oyster Crassostrea virginica along the east coast of the United States. Very few data have been collected, however, on the abundance of the parasite in environmental waters, limiting our understanding of P. marinus transmission dynamics. Real-time PCR assays with SybrGreen I as a label for detection were developed in this study for quantification of P. marinus in environmental waters with P. marinus species specific primers and of Perkinsus spp. with Perkinsus genus-specific primers. Detection of DNA concentrations as low as the equivalent of 3.3 x 10(-2) cell per 10-microl reaction mixture was obtained by targeting the multicopy internal transcribed spacer region of the genome. To obtain reliable target quantification from environmental water samples, removal of PCR inhibitors and efficient DNA recovery were two major concerns. A DNA extraction kit designed for tissues and another designed for stool samples were tested on environmental and artificial seawater (ASW) samples spiked with P. marinus cultured cells. The stool kit was significantly more efficient than the tissue kit at removing inhibitors from environmental water samples. With the stool kit, no significant difference in the quantified target concentrations was observed between the environmental and ASW samples. However, with the spiked ASW samples, the tissue kit demonstrated more efficient DNA recovery. Finally, by performing three elutions of DNA from the spin columns, which were combined prior to target quantification, variability of DNA recovery from different samples was minimized and more reliable real-time PCR quantification was accomplished. PMID- 15528526 TI - Differentiation of Micromonospora isolates from a coastal sediment in Wales on the basis of Fourier transform infrared spectroscopy, 16S rRNA sequence analysis, and the amplified fragment length polymorphism technique. AB - A number of actinomycetes isolates were recovered from coastal sediments in Aberystwyth (Wales, United Kingdom) with standard isolation techniques. Most of them were putatively assigned to the genera Streptomyces and Micromonospora on the basis of their morphological characteristics, and there appeared to be no difference whether the isolation media contained distilled water or seawater. A group of 20 Micromonospora isolates was selected to undergo further polyphasic taxonomic investigation. Three approaches were used to analyze the diversity of these isolates, 16S rDNA sequencing, fluorescent amplified fragment length polymorphism (AFLP), and Fourier transform infrared spectroscopy (FT-IR). The 16S rDNA sequence analysis confirmed that all of these isolates should be classified to the genus Micromonospora, and they were analyzed with a group of other Micromonospora 16S rDNA sequences available from the Ribosomal Database Project. The relationships of the 20 isolates were observed after hierarchical clustering, and almost identical clusters were obtained with these three techniques. This has obvious implications for high-throughput screening for novel actinomycetes because FT-IR spectroscopy, which is a rapid and reliable whole-organism fingerprinting method, can be applied as a very useful dereplication tool to indicate which environmental isolates have been cultured previously. PMID- 15528527 TI - Sequence and expression analysis of the ompA gene of Rickettsia peacockii, an endosymbiont of the Rocky Mountain wood tick, Dermacentor andersoni. AB - The transmission dynamics of Rocky Mountain spotted fever in Montana appears to be regulated by Rickettsia peacockii, a tick symbiotic rickettsia that interferes with transmission of virulent Rickettsia rickettsii. To elucidate the molecular relationships between the two rickettsiae and glean information on how to possibly exploit this interference phenomenon, we studied a major rickettsial outer membrane protein gene, ompA, presumed to be involved in infection and pathogenesis of spotted fever group rickettsiae (SFGR) but which is not expressed in the symbiont. Based on PCR amplification and DNA sequence analysis of the SFGR ompA gene, we demonstrate that R. peacockii is the most closely related of all known SFGR to R. rickettsii. We show that R. peacockii, originally described as East Side agent in Dermacentor andersoni ticks from the east side of the Bitterroot Valley in Montana, is still present in that tick population as well as in D. andersoni ticks collected at two widely separated locations in Colorado. The ompA genes of R. peacockii from these locations share three identical premature stop codons and a weakened ribosome binding site consensus sequence relative to ompA of R. rickettsii. The R. peacockii ompA promoter closely resembles that of R. rickettsii and is functional based on reverse transcription PCR results. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting showed that OmpA translation products were not detected in cultured tick cells infected with R. peacockii. Double immunolabeling studies revealed actin tail structures in tick cells infected with R. rickettsii strain Hlp#2 but not in cells infected with R. peacockii. PMID- 15528528 TI - Postprocessing in vitro digestion challenge to evaluate survival of Escherichia coli O157:H7 in fermented dry sausages. AB - Fermented dry sausages, inoculated with Escherichia coli O157:H7 during batter preparation, were submitted to an in vitro digestion challenge to evaluate the extent to which passage through the human gastrointestinal tract could inactivate the pathogenic cells, previously stressed by the manufacturing process. The numbers of surviving E. coli O157:H7 cells remained constant after a 1-min exposure of the finely chopped sausage to synthetic saliva or during the following 120-min exposure to synthetic gastric juice at an initial pH of 2.0. However, significant (P < or = 0.05) growth of the pathogen (1.03 to 2.16 log10 CFU/g) was observed in a subsequent 250-min exposure to a synthetic pancreatic juice at pH 8.0. In a different set of experiments, fractions from the gastric suspension were transferred into the synthetic pancreatic juice at 30-min intervals to mimic the dynamics of gastric emptying. Concurrently, the pH of the remaining gastric fluid was reduced to 3.0, 2.5, and 2.0 to simulate the gradual reacidification of the stomach contents after the initial buffering effect resulting from meal ingestion. Under these new conditions, pathogen growth during pancreatic challenge was observed for the first few fractions released from the stomach (90 min of exposure [pH 2.5]), but growth was no longer possible in the fractions submitted to the most severe gastric challenge (120 min of exposure [pH < 2.2]). PMID- 15528529 TI - Heavy-metal stress and developmental patterns of arbuscular mycorrhizal fungi. AB - The rate of global deposition of Cd, Pb, and Zn has decreased over the past few decades, but heavy metals already in the soil may be mobilized by local and global changes in soil conditions and exert toxic effects on soil microorganisms. We examined in vitro effects of Cd, Pb, and Zn on critical life stages in metal sensitive ecotypes of arbuscular mycorrhizal (AM) fungi, including spore germination, presymbiotic hyphal extension, presymbiotic sporulation, symbiotic extraradical mycelium expansion, and symbiotic sporulation. Despite long-term culturing under the same low-metal conditions, two species, Glomus etunicatum and Glomus intraradices, had different levels of sensitivity to metal stress. G. etunicatum was more sensitive to all three metals than was G. intraradices. A unique response of increased presymbiotic hyphal extension occurred in G. intraradices exposed to Cd and Pb. Presymbiotic hyphae of G. intraradices formed presymbiotic spores, whose initiation was more affected by heavy metals than was presymbiotic hyphal extension. In G. intraradices grown in compartmentalized habitats with only a portion of the extraradical mycelium exposed to metal stress, inhibitory effects of elevated metal concentrations on symbiotic mycelial expansion and symbiotic sporulation were limited to the metal-enriched compartment. Symbiotic sporulation was more sensitive to metal exposure than symbiotic mycelium expansion. Patterns exhibited by G. intraradices spore germination, presymbiotic hyphal extension, symbiotic extraradical mycelium expansion, and sporulation under elevated metal concentrations suggest that AM fungi may be able to survive in heavy metal-contaminated environments by using a metal avoidance strategy. PMID- 15528530 TI - Dynamics of multiple lin gene expression in Sphingomonas paucimobilis B90A in response to different hexachlorocyclohexane isomers. AB - Sphingomonas paucimobilis B90A is able to degrade the alpha-, beta-, gamma-, and delta-isomers of hexachlorocyclohexane (HCH). It contains the genes linA, linB, linC, linD, linE, and linR, which have been implicated in HCH degradation. In this study, dynamic expression of the lin genes was measured in chemostat-grown S. paucimobilis B90A by RNA dot blot hybridization and real-time reverse transcriptase PCR upon exposure to a pulse of different HCH isomers. Irrespective of the addition of HCH, linA, linB, and linC were all expressed constitutively. In contrast, linD and linE were induced with alpha-HCH (2 mg/liter) and gamma-HCH (7 mg/liter). A sharp increase in mRNA levels for linD and linE was observed from 10 to 45 min after the addition of alpha- or gamma-HCH. Induction of linD and linE was not detectable upon the addition of 0.7 mg of gamma-HCH per liter, although the compound was degraded by the cells. The addition of beta-HCH (5 mg/liter) or delta-HCH (20 mg/liter) did not lead to linE and linD induction, despite the fact that 50% of the compounds were degraded. This suggests that degradation of beta- and delta-HCH proceeds by a different pathway than that of alpha- and gamma-HCH. PMID- 15528531 TI - Molecular characterization of Brevibacillus laterosporus and its potential use in biological control. AB - Thirty-three strains of Brevibacillus laterosporus, including three novel strains isolated from Brazilian soil samples, were examined for genetic variability by the use of different PCR-based methods. Molecular markers that could characterize bacterial strains with regards to their pathogenic potential were investigated. In addition, toxicity was assessed by the use of insects belonging to the orders Lepidoptera and Coleoptera and the mollusk Biomphalaria glabrata. Among the targets tested, Biomphalaria glabrata demonstrated the highest degree of sensitivity to B. laterosporus, with some strains inducing 90 to 100% mortality in snails aged 3 and 12 days posteclosion. Larvae of the coleopteron Anthonomus grandis were also susceptible, presenting mortality levels of between 33 and 63%. Toxicity was also noted towards the lepidopteron Anticarsia gemmatalis. In contrast, no mortality was recorded among test populations of Tenebrio molitor or Spodoptera frugiperda. The application of intergenic transcribed spacer PCR and BOX-PCR generated 15 and 17 different genotypes, respectively. None of the molecular techniques allowed the identification of a convenient marker that was associated with any entomopathogenic phenotype. However, a 1,078-bp amplicon was detected for all strains of B. laterosporus when a primer for amplification of the BOXA1R region was used. Similarly, a 900-bp amplicon was generated from all isolates by use of the primer OPA-11 for randomly amplified polymorphic DNA analysis. These amplicons were not detected for other phenotypically related Brevibacillus species, indicating that they represent markers that are specific for B. laterosporus, which may prove useful for the isolation and identification of new strains of this species. PMID- 15528532 TI - Pore size dependence on growth temperature is a common characteristic of the major outer membrane protein OprF in psychrotrophic and mesophilic Pseudomonas species. AB - Pseudomonas species adapt well to hostile environments, which are often subjected to rapid variations. In these bacteria, the outer membrane plays an important role in the sensing of environmental conditions such as temperature. In previous studies, it has been shown that in the psychrotrophic strain P. fluorescens MF0, the major porin OprF changes its channel size according to the growth conditions and could affect outer membrane permeability. Studies of the channel-forming properties of OprFs from P. putida 01G3 and P. aeruginosa PAO1 in planar lipid bilayers generated similar results. The presence of a cysteine- or proline-rich cluster in the central linker region is not essential for channel size modulations. These findings suggest that OprF could adopt two alternative conformations in the outer membrane and that folding is thermoregulated. In contrast, no difference according to growth temperature was observed for structurally different outer membrane proteins, such as OprE3 from the Pseudomonas OprD family of specific porins. Our results are consistent with the fact that the decrease in channel size observed at low growth temperature is a particular feature of the OprF porin in various psychrotrophic and mesophilic Pseudomonas species isolated from diverse ecological niches. The ability to reduce outer membrane permeability at low growth temperature could provide these bacteria with adaptive advantages. PMID- 15528533 TI - Life history implications of rRNA gene copy number in Escherichia coli. AB - The role of the rRNA gene copy number as a central component of bacterial life histories was studied by using strains of Escherichia coli in which one or two of the seven rRNA operons (rrnA and/or rrnB) were deleted. The relative fitness of these strains was determined in competition experiments in both batch and chemostat cultures. In batch cultures, the decrease in relative fitness corresponded to the number of rRNA operons deleted, which could be accounted for completely by increased lag times and decreased growth rates. The magnitude of the deleterious effect varied with the environment in which fitness was measured: the negative consequences of rRNA operon deletions increased under culture conditions permitting more-rapid growth. The rRNA operon deletion strains were not more effective competitors under the regimen of constant, limited resources provided in chemostat cultures. Enhanced fitness in chemostat cultures would have suggested a simple tradeoff in which deletion strains grew faster (due to more efficient resource utilization) under resource limitation. The contributions of growth rate, lag time, Ks, and death rate to the fitness of each strain were verified through mathematical simulation of competition experiments. These data support the hypothesis that multiple rRNA operons are a component of bacterial life history and that they confer a selective advantage permitting microbes to respond quickly and grow rapidly in environments characterized by fluctuations in resource availability. PMID- 15528534 TI - Novel sulfonolipid in the extremely halophilic bacterium Salinibacter ruber. AB - Salinibacter ruber is an extremely halophilic bacterium, phylogenetically affiliated with the Flavobacterium/Cytophaga branch of the domain Bacteria. Electrospray mass analyses (negative ion) of the total lipid extract of a pure culture of S. ruber shows a characteristic peak at m/z 660 as the most prominent peak in the high-mass range of the spectrum. A novel sulfonolipid, giving rise to the molecular ion [M-H]- of m/z 660, has been identified. The sulfonolipid isolated and purified by thin-layer chromatography was shown by chemical degradation, mass spectrometry, infrared spectroscopy, and nuclear magnetic resonance analysis to have the structure 2-carboxy-2-amino-3-O-(13' methyltetradecanoyl)-4-hydroxy-18-methylnonadec-5-ene-1-sulfonic acid. This lipid represents about 10% of total cellular lipids, and it appears to be a structural variant of the sulfonolipids found as main components of the cell envelope of gliding bacteria of the genus Cytophaga and closely related genera (W. Godchaux and E. R. Leadbetter, J. Bacteriol. 153:1238-1246, 1983) and of diatoms (R. Anderson, M. Kates, and B. E. Volcani, Biochim. Biophys. Acta 528:89-106, 1978). Since this sulfonolipid has never been observed in any other extreme halophilic microorganism, we consider the peak at m/z 660 the lipid signature of Salinibacter. This study suggests that this novel sulfonolipid may be used as a chemotaxonomic marker for the detection of Salinibacter within the halophilic microbial community in saltern crystallizer ponds and other hypersaline environments. PMID- 15528535 TI - Morphotypic conversion in Listeria monocytogenes biofilm formation: biological significance of rough colony isolates. AB - Adherence to a stainless steel surface selected isolates of Listeria monocytogenes with enhanced surface colonization abilities and a change in phenotype from the common smooth colony morphology to a succession of rough colony morphotypes. Growth in broth culture of the best-adapted, surface colonizing rough colony morphotype gave a smooth colony revertant. Comparative analysis revealed that the smooth and rough variants had similar phenotypic and biochemical characteristics (e.g., identical growth rates and tolerances to antibiotics and environmental stressors). Rough colony isolates, however, failed to coordinate motility or induce autolysis. The defect in autolysis of rough colony isolates, which involved impaired cellular localization of several peptidoglycan-degrading enzymes, including cell wall hydrolase A (CwhA), suggested a link to a secretory pathway defect. The genetic basis for the impairment was studied at the level of the accessory secretory pathway component SecA2. DNA sequencing of the secA2 gene in smooth and rough colony isolates found no mutations in the coding or promoter regions. Analysis of SecA2 expression with an integrated secA2-FLAG tag construct found the protein to be upregulated in the rough and revertant backgrounds compared to the parental smooth colony isolate. A compensatory mechanism involving the SecA2 secretion pathway components is postulated to control smooth to rough interconversion of L. monocytogenes. Such phenotypic variation may enhance the ability of this opportunistic pathogen to colonize environments as diverse as processing surfaces, food products, and animal hosts. PMID- 15528536 TI - Use of cell culture-PCR assay based on combination of A549 and BGMK cell lines and molecular identification as a tool to monitor infectious adenoviruses and enteroviruses in river water. AB - Viral contamination in environmental samples can be underestimated because a single cell line might reproduce only some enteric viruses and some enteric viruses do not exhibit apparent cytopathic effects in cell culture. To overcome this problem, we evaluated a cell culture-PCR assay based on a combination of A549 and Buffalo green monkey kidney (BGMK) cell lines as a tool to monitor infectious adenoviruses and enteroviruses in river water. Water samples were collected 10 times at each of four rivers located in Gyeonggi Province, South Korea, and then cultured on group 1 cells (BGMK cells alone) and group 2 cells (BGMK and A549 cells). Reverse transcription and multiplex PCR were performed, followed by phylogenetic analysis of the amplicons. Thirty (75.0%) of the 40 samples were positive for viruses based on cell culture, and the frequency of positive samples grown on group 2 cells (65.0%) was higher than the frequency of positive samples grown on group 1 cells (50.0%). The number of samples positive for adenoviruses was higher with A549 cells (13 samples) than with BGMK cells (one sample); the numbers of samples positive for enteroviruses were similar with both types of cells. By using phylogenetic analysis, adenoviral amplicons were grouped into subgenera A, C, D, and F, and enteroviral amplicons were grouped into coxsackieviruses B3 and B4 and echoviruses 6, 7, and 30, indicating that A549 and BGMK cells were suitable for recovering a wide range of adenoviral and enteroviral types. The cell culture-PCR assay with a combination of A549 and BGMK cells and molecular identification could be a useful tool for monitoring infectious adenoviruses and enteroviruses in aquatic environments. PMID- 15528537 TI - Benthic and pelagic viral decay experiments: a model-based analysis and its applicability. AB - The viral decay in sediments, that is, the decrease in benthic viral concentration over time, was recorded after inhibiting the production of new viruses. Assuming that the viral abundance in an aquatic system remains constant and that viruses from lysed bacterial cells replace viruses lost by decay, the decay of viral particles can be used as a measure of viral production. Decay experiments showed that this approach is a useful tool to assess benthic viral production. However, the time course pattern of the decay experiments makes their interpretation difficult, regardless of whether viral decay is determined in the water column or in sediments. Different curve-fitting approaches (logarithmic function, power function, and linear regression) to describe the time course of decay experiments found in the literature are used in the present study and compared to a proposed "exponential decay" model based on the assumption that at any moment the decay is proportional to the amount of viruses present. Thus, an equation of the form dVA/dt = -k x VA leading to the time-integrated form VAt = VA0 x e(-k x t) was used, where k represents the viral decay rate (h(-1)), VAt is the viral abundance (viral particles ml(-1)) at time t (h), and VA0 is the initial viral abundance (viral particles ml(-1)). This approach represents the best solution for an accurate curve fitting based on a mathematical model for a realistic description of viral decay occurring in aquatic systems. Decay rates ranged from 0.0282 to 0.0696 h(-1) (mean, 0.0464 h(-1)). Additionally, a mathematical model is presented that enables the quantification of the viral control of bacterial production. The viral impact on bacteria based on decay rates calculated from the different mathematical approaches varied widely within one and the same decay experiment. A comparison of the viral control of bacterial production in different aquatic environments is, therefore, improper when different mathematical formulas are used to interpret viral decay experiments. PMID- 15528538 TI - Identification and functional analysis of two aromatic-ring-hydroxylating dioxygenases from a sphingomonas strain that degrades various polycyclic aromatic hydrocarbons. AB - In this study, the enzymes involved in polycyclic aromatic hydrocarbon (PAH) degradation in the chrysene-degrading organism Sphingomonas sp. strain CHY-1 were investigated. [14C]chrysene mineralization experiments showed that PAH-grown bacteria produced high levels of chrysene-catabolic activity. One PAH-induced protein displayed similarity with a ring-hydroxylating dioxygenase beta subunit, and a second PAH-induced protein displayed similarity with an extradiol dioxygenase. The genes encoding these proteins were cloned, and sequence analysis revealed two distinct loci containing clustered catabolic genes with strong similarities to corresponding genes found in Novosphingobium aromaticivorans F199. In the first locus, two genes potentially encoding a terminal dioxygenase component, designated PhnI, were followed by a gene coding for an aryl alcohol dehydrogenase (phnB). The second locus contained five genes encoding an extradiol dioxygenase (phnC), a ferredoxin (phnA3), another oxygenase component (PhnII), and an isomerase (phnD). PhnI was found to be capable of converting several PAHs, including chrysene, to the corresponding dihydrodiols. The activity of PhnI was greatly enhanced upon coexpression of genes encoding a ferredoxin (phnA3) and a reductase (phnA4). Disruption of the phnA1a gene encoding the PhnI alpha subunit resulted in a mutant strain that had lost the ability to grow on PAHs. The recombinant PhnII enzyme overproduced in Escherichia coli functioned as a salicylate 1-hydroxylase. PhnII also used methylsalicylates and anthranilate as substrates. Our results indicated that a single enzyme (PhnI) was responsible for the initial attack of a range of PAHs, including chrysene, in strain CHY-1. Furthermore, the conversion of salicylate to catechol was catalyzed by a three component oxygenase unrelated to known salicylate hydroxylases. PMID- 15528539 TI - Denaturing gradient gel electrophoretic analysis of ammonia-oxidizing bacterial community structure in the lower Seine River: impact of Paris wastewater effluents. AB - The Seine River is strongly affected by the effluents from the Acheres wastewater treatment plant (WWTP) downstream of the city of Paris. We have shown that the effluents introduce large amounts of ammonia and inoculate the receiving medium with nitrifying bacteria. The aim of the present study was to investigate the diversity of the ammonia-oxidizing bacterial population by identifying autochthonous bacteria from upstream and/or allochthonous ammonia-oxidizing bacteria from the WWTP effluents. Measurements of potential nitrifying activity, competitive PCR, and denaturing gradient gel electrophoresis (DGGE) of 16S ribosomal DNA fragments specific to ammonia-oxidizing bacteria (AOB) were used to explore the succession and shifts of the ammonia-oxidizing community in the lower Seine River and to analyze the temporal and spatial functioning of the system at several different sampling dates. A major revelation was the stability of the patterns. The CTO primers used in this study (G. A. Kowalchuk, J. R. Stephen, W. D. Boer, J. I. Prosser, T. M. Embley, and J. W. Woldendorp, Appl. Environ. Microbiol. 63:1489-1497, 1997) were shown not to be completely specific to AOB of the beta subclass of Proteobacteria. We further demonstrated that when DGGE patterns are interpreted, all the different bands must be sequenced, as one major DGGE band proved to be affiliated with a group of non-AOB in the beta subclass of Proteobacteria. The majority of AOB (75 to 90%) present in the lower Seine river downstream of the effluent output belong to lineage 6a, represented by Nitrosomonas oligotropha- and Nitrosomonas ureae-like bacteria. This dominant lineage was represented by three bands on the DGGE gel. The major lineage-6a AOB species, introduced by the WWTP effluents, survived and might have grown in the receiving medium far downstream, in the estuary; it represented about 40% of the whole AOB population. The other two species belonging to lineage 6a seem to be autochthonous bacteria. One of them developed a few kilometers downstream of the WWTP effluent input in an ammonia-enriched environment, and the other appeared in the freshwater part of the estuary and was apparently more adapted to estuarine conditions, i.e., an increase in the amount of suspended matter, a low ammonia concentration, and high turnover of organic matter. The rest of the AOB population was represented in equal proportions by Nitrosospira- and Nitrosococcus mobilis-like species. PMID- 15528540 TI - DNA Macroarray profiling of Lactococcus lactis subsp. lactis IL1403 gene expression during environmental stresses. AB - This report describes the use of an oligonucleotide macroarray to profile the expression of 375 genes in Lactococcus lactis subsp. lactis IL1403 during heat, acid, and osmotic stress. A set of known stress-associated genes in IL1403 was used as the internal control on the array. Every stress response was accurately detected using the macroarray, compared to data from previous reports. As a group, the expression patterns of the investigated metabolic genes were significantly altered by heat, acid, and osmotic stresses. Specifically, 13 to 18% of the investigated genes were differentially expressed in each of the environmental stress treatments. Interestingly, the methionine biosynthesis pathway genes (metA-metB1 and metB2-cysK) were induced during heat shock, but methionine utilization genes, such as metK, were induced during acid stress. These data provide a possible explanation for the differences between acid tolerance mechanisms of L. lactis strains IL1403 and MG1363 reported previously. Several groups of transcriptional responses were common among the stress treatments, such as repression of peptide transporter genes, including the opt operon (also known as dpp) and dtpT. Reduction of peptide transport due to environmental stress will have important implications in the cheese ripening process. Although stress responses in lactococci were extensively studied during the last decade, additional information about this bacterium was gained from the use of this metabolic array. PMID- 15528541 TI - Seasonal shedding of multiple Cryptosporidium genotypes in California ground squirrels (Spermophilus beecheyi). AB - Twelve percent of 853 California ground squirrels (Spermophilus beecheyi) from six different geographic locations in Kern County, Calif., were found to be shedding on average 44,482 oocysts g of feces(-1). The mean annual environmental loading rate of Cryptosporidium oocysts was 57,882 oocysts squirrel(-1) day(-1), with seasonal patterns of fecal shedding ranging from <10,000 oocysts squirrel( 1) day(-1) in fall, winter, and spring to levels of 2 x 10(5) oocysts squirrel( 1) day(-1) in summer. Juveniles were about twice as likely as adult squirrels to be infected and shed higher concentrations of oocysts than adults did, with particularly high levels of infection and shedding being found among juvenile male squirrels. Based on DNA sequencing of a portion of the 18S small-subunit rRNA gene, there existed three genotypes of Cryptosporidium species in these populations of squirrels (Sbey03a, Sbey03b, and Sbey03c; accession numbers AY462231 to AY462233, respectively). These unique DNA sequences were most closely related (96 to 97% homology) to porcine C. parvum (AF115377) and C. wrairi (AF115378). Inoculating BALB/c neonatal mice with up to 10,000 Sbey03b or Sbey03c fresh oocysts from different infected hosts did not produce detectable levels of infection, suggesting that this common genotype shed by California ground squirrels is not infectious for mice and may constitute a new species of Cryptosporidium. PMID- 15528542 TI - Changes in bacterioplankton composition under different phytoplankton regimens. AB - The results of empirical studies have revealed links between phytoplankton and bacterioplankton, such as the frequent correlation between chlorophyll a and bulk bacterial abundance and production. Nevertheless, little is known about possible links at the level of specific taxonomic groups. To investigate this issue, seawater microcosm experiments were performed in the northwestern Mediterranean Sea. Turbulence was used as a noninvasive means to induce phytoplankton blooms dominated by different algae. Microcosms exposed to turbulence became dominated by diatoms, while small phytoflagellates gained importance under still conditions. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments showed that changes in phytoplankton community composition were followed by shifts in bacterioplankton community composition, both as changes in the presence or absence of distinct bacterial phylotypes and as differences in the relative abundance of ubiquitous phylotypes. Sequencing of DGGE bands showed that four Roseobacter phylotypes were present in all microcosms. The microcosms with a higher proportion of phytoflagellates were characterized by four phylotypes of the Bacteroidetes phylum: two affiliated with the family Cryomorphaceae and two with the family Flavobacteriaceae. Two other Flavobacteriaceae phylotypes were characteristic of the diatom-dominated microcosms, together with one Alphaproteobacteria phylotype (Roseobacter) and one Gammaproteobacteria phylotype (Methylophaga). Phylogenetic analyses of published Bacteroidetes 16S rRNA gene sequences confirmed that members of the Flavobacteriaceae are remarkably responsive to phytoplankton blooms, indicating these bacteria could be particularly important in the processing of organic matter during such events. Our data suggest that quantitative and qualitative differences in phytoplankton species composition may lead to pronounced differences in bacterioplankton species composition. PMID- 15528543 TI - Bacterial diversity and function of aerobic granules engineered in a sequencing batch reactor for phenol degradation. AB - Aerobic granules are self-immobilized aggregates of microorganisms and represent a relatively new form of cell immobilization developed for biological wastewater treatment. In this study, both culture-based and culture-independent techniques were used to investigate the bacterial diversity and function in aerobic phenol- degrading granules cultivated in a sequencing batch reactor. Denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA genes demonstrated a major shift in the microbial community as the seed sludge developed into granules. Culture isolation and DGGE assays confirmed the dominance of beta Proteobacteria and high-G+C gram-positive bacteria in the phenol-degrading aerobic granules. Of the 10 phenol-degrading bacterial strains isolated from the granules, strains PG-01, PG-02, and PG-08 possessed 16S rRNA gene sequences that matched the partial sequences of dominant bands in the DGGE fingerprint belonging to the aerobic granules. The numerical dominance of strain PG-01 was confirmed by isolation, DGGE, and in situ hybridization with a strain-specific probe, and key physiological traits possessed by PG-01 that allowed it to outcompete and dominate other microorganisms within the granules were then identified. This strain could be regarded as a functionally dominant strain and may have contributed significantly to phenol degradation in the granules. On the other hand, strain PG-08 had low specific growth rate and low phenol degradation ability but showed a high propensity to autoaggregate. By analyzing the roles played by these two isolates within the aerobic granules, a functional model of the microbial community within the aerobic granules was proposed. This model has important implications for rationalizing the engineering of ecological systems. PMID- 15528544 TI - Poly(3-hydroxybutyrate) biosynthesis in the biofilm of Alcaligenes eutrophus, using glucose enzymatically released from pulp fiber sludge. AB - Glucose, enzymatically released from pulp fiber sludge, was combined with inorganic salts and used as a growth medium for Alcaligenes eutrophus, a gram negative strain producing poly(3-hydroxybutyrate) (PHB). By controlling the concentrations of the inorganic salts in the growth medium, almost 78% of the cell mass was converted to pure PHB. Efforts were made to find conditions for bacterial growth in the form of a biofilm on a cheap and reusable carrier. A number of positively charged carriers were tested, and the anion exchanger DEAE Sephadex A-25 was chosen as a microcarrier for packed-bed biofilm cultures of A. eutrophus. Conditions for attachment, growth, and detachment were established. Biofilm formation on the microcarrier is strongly dependent on the ionic strength of the attachment medium. In order to achieve formation of the biofilm and its recovery from the microcarrier, the ionic strengths of the attachment and the detachment media were varied. Low ionic strength was tested for attachment, and high ionic strength was tested for detachment. Although biofilm formation in the packed-bed reactor is limited, the volumetric yield of cells based on the void volume of the packed bed is comparable with the batch culture yield. PMID- 15528545 TI - Interaction and transmission of two Borrelia burgdorferi sensu stricto strains in a tick-rodent maintenance system. AB - In the northeastern United States, the Lyme disease agent, Borrelia burgdorferi sensu stricto, is maintained by enzoonotic transmission, cycling between white footed mice (Peromyscus leucopus) and black-legged ticks (Ixodes scapularis). B. burgdorferi sensu stricto is genetically variable and has been divided into three major genotypes based on 16S-23S ribosomal DNA spacer (RST) analysis. To better understand how genetic differences in B. burgdorferi sensu stricto may influence transmission dynamics in nature, we investigated the interaction between an RST1 and an RST3 strain in a laboratory system with P. leucopus mice and I. scapularis ticks. Two groups of mice were infected with either BL206 (RST1) or B348 (RST3). Two weeks later, experimental mice were challenged with the opposite strain, while control mice were challenged with the same strain as that used for the primary infection. The transmission of BL206 and B348 from infected mice was then determined by xenodiagnosis with uninfected larval ticks at weekly intervals for 42 days. Mice in both experimental groups were permissive for infection with the second strain and were able to transmit both strains to the xenodiagnostic ticks. However, the overall transmission efficiencies of BL206 and B348 were significantly different. BL206 was more efficiently transmitted than B348 to xenodiagnostic ticks. Significantly fewer double infections than expected were detected in xenodiagnostic ticks. The results suggest that some B. burgdorferi sensu stricto strains, such as BL206, may be preferentially maintained in transmission cycles between ticks and white-footed mice. Other strains, such as B348, may be more effectively maintained in different tick-vertebrate transmission cycles. PMID- 15528546 TI - Engineering of chimeric class II polyhydroxyalkanoate synthases. AB - PHA synthase is a key enzyme involved in the biosynthesis of polyhydroxyalkanoates (PHAs). Using a combinatorial genetic strategy to create unique chimeric class II PHA synthases, we have obtained a number of novel chimeras which display improved catalytic properties. To engineer the chimeric PHA synthases, we constructed a synthetic phaC gene from Pseudomonas oleovorans (phaC1Po) that was devoid of an internal 540-bp fragment. Randomly amplified PCR products (created with primers based on conserved phaC sequences flanking the deleted internal fragment) were generated using genomic DNA isolated from soil and were substituted for the 540-bp internal region. The chimeric genes were expressed in a PHA-negative strain of Ralstonia eutropha, PHB(-)4 (DSM 541). Out of 1,478 recombinant clones screened for PHA production, we obtained five different chimeric phaC1Po genes that produced more PHA than the native phaC1Po. Chimeras S1-71, S4-8, S5-58, S3-69, and S3-44 exhibited 1.3-, 1.4-, 2.0-, 2.1-, and 3.0-fold-increased levels of in vivo activity, respectively. All of the mutants mediated the synthesis of PHAs with a slightly increased molar fraction of 3-hydroxyoctanoate; however, the weight-average molecular weights (Mw) of the PHAs in all cases remained almost the same. Based upon DNA sequence analyses, the various phaC fragments appear to have originated from Pseudomonas fluorescens and Pseudomonas aureofaciens. The amino acid sequence analyses showed that the chimeric proteins had 17 to 20 amino acid differences from the wild-type phaC1Po, and these differences were clustered in the same positions in the five chimeric clones. A threading model of PhaC1Po, developed based on homology of the enzyme to the Burkholderia glumae lipase, suggested that the amino acid substitutions found in the active chimeras were located mostly on the protein model surface. Thus, our combinatorial genetic engineering strategy proved to be broadly useful for improving the catalytic activities of PHA synthase enzymes. PMID- 15528547 TI - Ultrastructural alterations of Erwinia carotovora subsp. atroseptica caused by treatment with aluminum chloride and sodium metabisulfite. AB - Aluminum and bisulfite salts inhibit the growth of several fungi and bacteria, and their application effectively controls potato soft rot caused by Erwinia carotovora. In an effort to understand their inhibitory action, ultrastructural changes in Erwinia carotovora subsp. atroseptica after exposure (0 to 20 min) to different concentrations (0.05, 0.1, and 0.2 M) of these salts were examined by using transmission electron microscopy. Plasma membrane integrity was evaluated by using the SYTOX Green fluorochrome that penetrates only cells with altered membranes. Bacteria exposed to all aluminum chloride concentrations, especially 0.2 M, exhibited loosening of the cell walls, cell wall rupture, cytoplasmic aggregation, and an absence of extracellular vesicles. Sodium metabisulfite caused mainly a retraction of plasma membrane and cellular voids which were more pronounced with increasing concentration. Bacterial mortality was closely associated with SYTOX stain absorption when bacteria were exposed to either a high concentration (0.2 M) of aluminum chloride or prolonged exposure (20 min) to 0.05 M aluminum chloride or to a pH of 2.5. Bacteria exposed to lower concentrations of aluminum chloride (0.05 and 0.1 M) for 10 min or less, or to metabisulfite at all concentrations, did not exhibit significant stain absorption, suggesting that no membrane damage occurred or it was too weak to allow the penetration of the stain into the cell. While mortality caused by aluminum chloride involves membrane damage and subsequent cytoplasmic aggregation, sulfite exerts its effect intracellularly; it is transported across the membrane by free diffusion of molecular SO2 with little damage to the cellular membrane. PMID- 15528548 TI - Visualization of differential gene expression by improved cyan fluorescent protein and yellow fluorescent protein production in Bacillus subtilis. AB - The distinguishable cyan and yellow fluorescent proteins (CFP and YFP) enable the simultaneous in vivo visualization of different promoter activities. Here, we report new cloning vectors for the construction of cfp and yfp fusions in Bacillus subtilis. By extending the N-terminal portions of previously described CFP and YFP variants, 20- to 70-fold-improved fluorescent-protein production was achieved. Probably, the addition of sequences encoding the first eight amino acids of the N-terminal part of ComGA of B. subtilis overcomes the slow translation initiation that is provoked by the eukaryotic codon bias present in the original cfp and yfp genes. Using these new vectors, we demonstrate that, within an isogenic population of sporulating B. subtilis cells, expression of the abrB and spoIIA genes is distinct in individual cells. PMID- 15528549 TI - Saccharomyces cerevisiae engineered for xylose metabolism exhibits a respiratory response. AB - Native strains of Saccharomyces cerevisiae do not assimilate xylose. S. cerevisiae engineered for d-xylose utilization through the heterologous expression of genes for aldose reductase (XYL1), xylitol dehydrogenase (XYL2), and d-xylulokinase (XYL3 or XKS1) produce only limited amounts of ethanol in xylose medium. In recombinant S. cerevisiae expressing XYL1, XYL2, and XYL3, mRNA transcript levels for glycolytic, fermentative, and pentose phosphate enzymes did not change significantly on glucose or xylose under aeration or oxygen limitation. However, expression of genes encoding the tricarboxylic acid cycle, respiration enzymes (HXK1, ADH2, COX13, NDI1, and NDE1), and regulatory proteins (HAP4 and MTH1) increased significantly when cells were cultivated on xylose, and the genes for respiration were even more elevated under oxygen limitation. These results suggest that recombinant S. cerevisiae does not recognize xylose as a fermentable carbon source and that respiratory proteins are induced in response to cytosolic redox imbalance; however, lower sugar uptake and growth rates on xylose might also induce transcripts for respiration. A petite respiration deficient mutant (rho degrees ) of the engineered strain produced more ethanol and accumulated less xylitol from xylose. It formed characteristic colonies on glucose, but it did not grow on xylose. These results are consistent with the higher respiratory activity of recombinant S. cerevisiae when growing on xylose and with its inability to grow on xylose under anaerobic conditions. PMID- 15528550 TI - Intracellular proliferation of Legionella pneumophila in Hartmannella vermiformis in aquatic biofilms grown on plasticized polyvinyl chloride. AB - The need for protozoa for the proliferation of Legionella pneumophila in aquatic habitats is still not fully understood and is even questioned by some investigators. This study shows the in vivo growth of L. pneumophila in protozoa in aquatic biofilms developing at high concentrations on plasticized polyvinyl chloride in a batch system with autoclaved tap water. The inoculum, a mixed microbial community including indigenous L. pneumophila originating from a tap water system, was added in an unfiltered as well as filtered (cellulose nitrate, 3.0-microm pore size) state. Both the attached and suspended biomasses were examined for their total amounts of ATP, for culturable L. pneumophila, and for their concentrations of protozoa. L. pneumophila grew to high numbers (6.3 log CFU/cm2) only in flasks with an unfiltered inoculum. Filtration obviously removed the growth-supporting factor, but it did not affect biofilm formation, as determined by measuring ATP. Cultivation, direct counting, and 18S ribosomal DNA targeted PCR with subsequent sequencing revealed the presence of Hartmannella vermiformis in all flasks in which L. pneumophila multiplied and also when cycloheximide had been added. Fluorescent in situ hybridization clearly demonstrated the intracellular growth of L. pneumophila in trophozoites of H. vermiformis, with 25.9% +/- 10.5% of the trophozoites containing L. pneumophila on day 10 and >90% containing L. pneumophila on day 14. Calculations confirmed that intracellular growth was most likely the only way for L. pneumophila to proliferate within the biofilm. Higher biofilm concentrations, measured as amounts of ATP, gave higher L. pneumophila concentrations, and therefore the growth of L. pneumophila within engineered water systems can be limited by controlling biofilm formation. PMID- 15528551 TI - Influence of an oyster reef on development of the microbial heterotrophic community of an estuarine biofilm. AB - We characterized microbial biofilm communities developed over two very closely located but distinct benthic habitats in the Pensacola Bay estuary using two complementary cultivation-independent molecular techniques. Biofilms were grown for 7 days on glass slides held in racks 10 to 15 cm over an oyster reef and an adjacent muddy sand bottom. Total biomass and optical densities of dried biofilms showed dramatic differences for oyster reef versus non-oyster reef biofilms. This study assessed whether the observed spatial variation was reflected in the heterotrophic prokaryotic species composition. Genomic biofilm DNA from both locations was isolated and served as a template to amplify 16S rRNA genes with universal eubacterial primers. Fluorescently labeled PCR products were analyzed by terminal restriction fragment length polymorphism, creating a genetic fingerprint of the composition of the microbial communities. Unlabeled PCR products were cloned in order to construct a clone library of 16S rRNA genes. Amplified ribosomal DNA restriction analysis was used to screen and define ribotypes. Partial sequences from unique ribotypes were compared with existing database entries to identify species and to construct phylogenetic trees representative of community structures. A pronounced difference in species richness and evenness was observed at the two sites. The biofilm community structure from the oyster reef setting had greater evenness and species richness than the one from the muddy sand bottom. The vast majority of the bacteria in the oyster reef biofilm were related to members of the gamma- and delta-subdivisions of Proteobacteria, the Cytophaga-Flavobacterium -Bacteroides cluster, and the phyla Planctomyces and Holophaga-Acidobacterium. The same groups were also present in the biofilm harvested at the muddy sand bottom, with the difference that nearly half of the community consisted of representatives of the Planctomyces phylum. Total species richness was estimated to be 417 for the oyster reef and 60 for the muddy sand bottom, with 10.5% of the total unique species identified being shared between habitats. The results suggest dramatic differences in habitat-specific microbial diversity that have implications for overall microbial diversity within estuaries. PMID- 15528552 TI - Identification of common subpopulations of non-sorbitol-fermenting, beta glucuronidase-negative Escherichia coli O157:H7 from bovine production environments and human clinical samples. AB - Non-sorbitol-fermenting, beta-glucuronidase-negative Escherichia coli O157:H7 strains are regarded as a clone complex, and populations from different geographical locations are believed to share a recent common ancestor. Despite their relatedness, high-resolution genotyping methods can detect significant genome variation among different populations. Phylogenetic analysis of high resolution genotyping data from these strains has shown that subpopulations from geographically unlinked continents can be divided into two primary phylogenetic lineages, termed lineage I and lineage II, and limited studies of the distribution of these lineages suggest there could be differences in their propensity to cause disease in humans or to be transmitted to humans. Because the genotyping methods necessary to discriminate the two lineages are tedious and subjective, these methods are not particularly suited for studying the large sets of strains that are required to systematically evaluate the ecology and transmission characteristics of these lineages. To overcome this limitation, we have developed a lineage-specific polymorphism assay (LSPA) that can readily distinguish between the lineage I and lineage II subpopulations. In the studies reported here, we describe the development of a six-marker test (LSPA-6) and its validation in a side-by-side comparison with octamer-based genome scanning. Analysis of over 1,400 O157:H7 strains with the LSPA-6 demonstrated that five genotypes comprise over 91% of the strains, suggesting that these subpopulations may be widespread. PMID- 15528553 TI - Relationship of Vibrio species infection and elevated temperatures to yellow blotch/band disease in Caribbean corals. AB - The bacterial and temperature factors leading to yellow blotch/band disease (YBD), which affects the major reef-building Caribbean corals Montastrea spp., have been investigated. Groups of bacteria isolated from affected corals and inoculated onto healthy corals caused disease signs similar to those of YBD. The 16S rRNA genes from these bacteria were sequenced and found to correspond to four Vibrio spp. Elevating the water temperature notably increased the rate of spread of YBD on inoculated corals and induced greater coral mortality. YBD-infected corals held at elevated water temperatures had 50% lower zooxanthella densities, 80% lower division rates, and a 75% decrease in chlorophyll a and c2 pigments compared with controls. Histological sections indicated that the algal pyrenoid was fragmented into separate segments, along with a reconfiguration and swelling of the zooxanthellae, as well as vacuolization. YBD does not appear to produce the same physiological response formerly observed in corals undergoing temperature-related bleaching. Evidence indicates that YBD affects primarily the symbiotic algae rather than coral tissue. PMID- 15528555 TI - Electric current-induced detachment of Staphylococcus epidermidis biofilms from surgical stainless steel. AB - Biomaterial-centered infections of orthopedic percutaneous implants are serious complications which can ultimately lead to osteomyelitis, with devastating effects on bone and surrounding tissues, especially since the biofilm mode of growth offers protection against antibiotics and since removal frequently is the only ultimate solution. Recently, it was demonstrated that as a possible pathway to prevent infections of percutaneous stainless steel implants, electric currents of 60 to 100 microA were effective at stimulating the detachment of initially adhering staphylococci from surgical stainless steel. However, initially adhering bacteria are known to adhere more reversibly than bacteria growing in the later stages of biofilm formation. Hence, the aim of this study was to examine whether a growing Staphylococcus epidermidis biofilm can be stimulated to detach from surgical stainless steel by the use of electric currents. In separate experiments, four currents, i.e., 60 and 100 microA of direct current (DC) and 60 and 100 microA of block current (50% duty cycle, 1 Hz), were applied for 360 min to stimulate the detachment of an S. epidermidis biofilm that had grown for 200 min. A 100-microA DC yielded 78% detachment, whereas a 100-microA block current under the same experimental conditions yielded only 31% detachment. The same trend was found for 60 microA, with 37% detachment for a DC and 24% for a block current. Bacteria remaining on the surface after the current application were less viable than they were prior to the current application, as demonstrated by confocal laser scanning microscopy. In conclusion, these results suggest that DCs are preferred for curing infections. PMID- 15528554 TI - Long-chain N-acyltyrosine synthases from environmental DNA. AB - The heterologous expression of DNA extracted directly from environmental samples (environmental DNA [eDNA]) in easily cultured hosts provides access to natural products produced by previously inaccessible microorganisms. When eDNA cosmid libraries were screened in Escherichia coli for antibacterially active clones, long-chain N-acyltyrosine-producing clones were found in every eDNA library. These apparently common natural products have not been previously described from screening extracts of cultured bacteria for biologically active natural products. Of the 11 long-chain N-acyl amino acid synthases (NASs) that were characterized, 10 are unique sequences. A predicted protein of previously unknown function from Nitrosomonas europaea, a gram-negative nitrifying beta-proteobacterium, is 14 to 37% identical to eDNA NASs. When cloned into E. coli, this open reading frame confers the production of long-chain N-acyltyrosines to the host and is therefore the first NAS from a cultured bacterium to be functionally characterized. Understanding the role that long-chain N-acyl amino acids play in soil microbial communities should now be feasible with the identification of a cultured organism that has the genetic capacity to produce these compounds. PMID- 15528556 TI - Indole-diterpene gene cluster from Aspergillus flavus. AB - Aflatrem is a potent tremorgenic mycotoxin produced by the soil fungus Aspergillus flavus and is a member of a large structurally diverse group of secondary metabolites known as indole-diterpenes. By using degenerate primers for conserved domains of fungal geranylgeranyl diphosphate synthases, we cloned two genes, atmG and ggsA (an apparent pseudogene), from A. flavus. Adjacent to atmG are two other genes, atmC and atmM. These three genes have 64 to 70% amino acid sequence similarity and conserved synteny with a cluster of orthologous genes, paxG, paxC, and paxM, from Penicillium paxilli which are required for indole diterpene biosynthesis. atmG, atmC, and atmM are coordinately expressed, with transcript levels dramatically increasing at the onset of aflatrem biosynthesis. A genomic copy of atmM can complement a paxM deletion mutant of P. paxilli, demonstrating that atmM is a functional homolog of paxM. Thus, atmG, atmC, and atmM are necessary, but not sufficient, for aflatrem biosynthesis by A. flavus. This provides the first genetic evidence for the biosynthetic pathway of aflatrem in A. flavus. PMID- 15528557 TI - Evaluation of the LIVE/DEAD BacLight kit for detection of extremophilic archaea and visualization of microorganisms in environmental hypersaline samples. AB - Extremophilic archaea were stained with the LIVE/DEAD BacLight kit under conditions of high ionic strength and over a pH range of 2.0 to 9.3. The reliability of the kit was tested with haloarchaea following permeabilization of the cells. Microorganisms in hypersaline environmental samples were detectable with the kit, which suggests its potential application to future extraterrestrial halites. PMID- 15528558 TI - New vector for efficient allelic replacement in naturally nontransformable, low GC-content, gram-positive bacteria. AB - A shuttle vector designated pMAD was constructed for quickly generating gene inactivation mutants in naturally nontransformable gram-positive bacteria. This vector allows, on X-Gal (5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) plates, a quick colorimetric blue-white discrimination of bacteria which have lost the plasmid, greatly facilitating clone identification during mutagenesis. The plasmid was used in Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus to efficiently construct mutants with or without an associated antibiotic resistance gene. PMID- 15528559 TI - Adhesion of Enterococcus faecalis in the nonculturable state to plankton is the main mechanism responsible for persistence of this bacterium in both lake and seawater. AB - The presence of enterococci in lake and seawater in an 18-month survey comparing molecular (PCR and quantitative PCR) and culture methods was evaluated, as well as the possibility that zooplankton could act as reservoirs for enterococci. Samples of both water and zooplankton were collected monthly from a Lake Garda site and an Adriatic Sea site. In lake water, the positive samples numbered 13 of 54 (24%) by culture and 32 of 54 (59%) when PCR was applied. In seawater, they numbered 0 of 51 by culture and 18 of 51 (35%) by PCR. Enterococci were found either totally bound to plankton or totally in water, depending on the presence or absence of plankton, respectively. These results clearly indicate that the PCR assay is a powerful tool for detecting fecal indicators and pathogens in the environment, thus providing a much more sensitive method than culture. PMID- 15528560 TI - Degradation of a nonylphenol single isomer by Sphingomonas sp. strain TTNP3 leads to a hydroxylation-induced migration product. AB - Sphingomonas sp. strain TTNP3 degrades 4(3',5'-dimethyl-3'-heptyl)-phenol and unidentified metabolites that were described previously. The chromatographic analyses of the synthesized reference compound and the metabolites led to their identification as 2(3',5'-dimethyl-3'-heptyl)-1,4-benzenediol. This finding indicates that the nonylphenol metabolism of this bacterium involves unconventional degradation pathways where an NIH shift mechanism occurs. PMID- 15528562 TI - In vitro laser ablation of natural marine biofilms. AB - We studied the efficiency of pulsed low-power laser irradiation of 532 nm from an Nd:YAG (neodymium-doped yttrium-aluminum-garnet) laser to remove marine biofilm developed on titanium and glass coupons. Natural biofilms with thicknesses of 79.4 +/- 27.8 microm (titanium) and 107.4 +/- 28.5 microm (glass) were completely disrupted by 30 s of laser irradiation (fluence, 0.1 J/cm2). Laser irradiation significantly reduced the number of diatoms and bacteria in the biofilm (paired t test; P < 0.05). The removal was better on titanium than on glass coupons. PMID- 15528561 TI - Use of transposon-transposase complexes to create stable insertion mutant strains of Francisella tularensis LVS. AB - Francisella tularensis is a highly virulent zoonotic bacterial pathogen capable of infecting numerous different mammalian species, including humans. Elucidation of the pathogenic mechanisms of F. tularensis has been hampered by a lack of tools to genetically manipulate this organism. Herein we describe the use of transposome complexes to create insertion mutations in the chromosome of the F. tularensis live vaccine strain (LVS). A Tn5-derived transposon encoding kanamycin resistance and lacking a transposase gene was complexed with transposase enzyme and transformed directly into F. tularensis LVS by electroporation. An insertion frequency of 2.6 x 10(-8) +/- 0.87 x 10(-8) per cell was consistently achieved using this method. There are 178 described Tn5 consensus target sites distributed throughout the F. tularensis genome. Twenty-two of 26 transposon insertions analyzed were within known or predicted open reading frames, but none of these insertions was associated with the Tn5 target site. Analysis of the insertions of sequentially passed strains indicated that the transposons were maintained stably at the initial insertion site after more than 270 generations. Therefore, transformation by electroporation of Tn5-based transposon-transposase complexes provided an efficient mechanism for generating random, stable chromosomal insertion mutations in F. tularensis. PMID- 15528563 TI - Detection and enumeration of Vibrio vulnificus in oysters from two estuaries along the southwest coast of India, using molecular methods. AB - This study was conducted to understand the seasonal distribution of Vibrio vulnificus in oysters from two estuaries and the effect of environmental factors on the abundance of V. vulnificus in tropical waters. V. vulnificus was detected in 56.6% of the samples tested by colony hybridization with an alkaline phosphatase-labeled oligonucleotide probe (VV-AP), and the counts ranged from <10/g during the summer months to 10(3)/g in the monsoon season at both sites. The density of V. vulnificus appeared to be controlled more by salinity than by temperature. A nested PCR used in this study detected V. vulnificus in 85% of the samples following 18 h of enrichment in alkaline peptone water. PMID- 15528564 TI - Complete type III secretion system of a mesophilic Aeromonas hydrophila strain. AB - We have investigated the existence and genetic organization of a functional type III secretion system (TTSS) in a mesophilic Aeromonas strain by initially using the Aeromonas hydrophila strain AH-3. We report for the first time the complete TTSS DNA sequence of an Aeromonas strain that comprises 35 genes organized in a similar disposition as that in Pseudomonas aeruginosa. Using several gene probes, we also determined the presence of a TTSS in clinical or environmental strains of different Aeromonas species: A. hydrophila, A. veronii, and A. caviae. By using one of the TTSS genes (ascV), we were able to obtain a defined insertion mutant in strain AH-3 (AH-3AscV), which showed reduced toxicity and virulence in comparison with the wild-type strain. Complementation of the mutant strain with a plasmid vector carrying ascV was fully able to restore the wild-type toxicity and virulence. PMID- 15528566 TI - Age-related decline in carriage of ampicillin-resistant Escherichia coli in young calves. AB - The presence of ampicillin-resistant Escherichia coli (Amp(r) E. coli) in the fecal flora of calves was monitored on a monthly basis in seven cohorts of calves. Calves were rapidly colonized by Amp(r) E. coli, with peak prevalence in cohort calves observed in the 4 months after the calves were born. The prevalence of calves yielding Amp(r) E. coli in cohorts consistently declined to low levels with increasing age of the calves (P < 0.001). PMID- 15528565 TI - Combined phospholipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis of bacterial diversity and physiological status in an intertidal microbial mat. AB - A combined lipid biomarker-16S rRNA gene denaturing gradient gel electrophoresis analysis was used to monitor changes in the physiological status, biomass, and microbial composition of a microbial mat. In the morning hours, an increase in the biomass of layers containing a high density of phototrophs and a decrease in the growth rate in the deep layers were observed. The combined approach also revealed differences in major groups of microorganisms, including green nonsulfur, gram-positive, and heterotrophic bacteria. PMID- 15528567 TI - Diversity of toxic shock syndrome toxin 1-positive Staphylococcus aureus isolates. AB - Staphylococcus aureus isolates from women with nasal, anal, or vaginal colonization were evaluated for population diversity by pulsed-field gel electrophoresis. Cluster analysis of restriction patterns revealed diversity indices of 0.89 and 0.99 for toxic shock syndrome toxin 1-positive and -negative isolates, respectively. Toxin-producing strains were isolated more frequently from the nares than from other sites. PMID- 15528568 TI - Selection of single-chain antibodies against the VP8* subunit of rotavirus VP4 outer capsid protein and their expression in Lactobacillus casei. AB - Single-chain antibodies (scFv) recognizing the VP8* fraction of rotavirus outer capsid and blocking rotavirus infection in vitro were isolated by phage display. Vectors for the extracellular expression in Lactobacillus casei of one of the scFv were constructed. L. casei was able to secrete active scFv to the growth medium, showing the potential of probiotic bacteria to be engineered to express molecules suitable for in vivo antirotavirus therapies. PMID- 15528569 TI - Acetate threshold concentrations suggest varying energy requirements during anaerobic respiration by Anaeromyxobacter dehalogenans. AB - Acetate threshold concentrations were determined under chlororespiring and Fe(III)-reducing conditions for Anaeromyxobacter dehalogenans strain 2CP-C. The acetate threshold concentrations measured were 69 +/- 4, 19 +/- 8, and <1 nM for chlororespiration, amorphous Fe(III) reduction, and Fe(III) citrate reduction, respectively. Residual DeltaG values of -75.4 kJ/mol of electrons for chlororespiration and -41.5 kJ/mol of electrons for amorphous Fe(III) reduction were calculated at the acetate threshold concentration. By comparing threshold concentrations for different metabolisms in a single organism, this study provides insight into the metabolic use of energy under different growth conditions. PMID- 15528570 TI - Verocytotoxin-producing Escherichia coli in wild birds and rodents in close proximity to farms. AB - Wild animals living close to cattle and pig farms (four each) were examined for verocytotoxin-producing Escherichia coli (VTEC; also known as Shiga toxin producing E. coli). The prevalence of VTEC among the 260 samples from wild animals was generally low. However, VTEC isolates from a starling (Sturnus vulgaris) and a Norway rat (Rattus norvegicus) were identical to cattle isolates from the corresponding farms with respect to serotype, virulence profile, and pulsed-field gel electrophoresis type. This study shows that wild birds and rodents may become infected from farm animals or vice versa, suggesting a possible role in VTEC transmission. PMID- 15528571 TI - Developing consensus in emergency medicine information technology. PMID- 15528572 TI - Emergency medicine information technology consensus conference: executive summary. PMID- 15528573 TI - Crafting information technology solutions, not experiments, for the emergency department. PMID- 15528574 TI - Keynote address: medical informatics and emergency medicine. AB - A personal look at some of the developments in practical clinical informatics over the past two decades, with discussion of several successful projects, including the National Center for Emergency Medicine Informatics, the Azyxxi system, Federal Project ER One, the Institutes for Innovation in Medicine, the Medical MediaLab, Project Sentinel, and others. Lessons learned, and hints and suggestions for future developers and informaticists. PMID- 15528575 TI - Where's the beef? The promise and the reality of clinical documentation. AB - Physician-generated emergency department clinical documentation (information obtained from clinician observations and summarized decision processes inclusive of all manner of electronic systems capturing, storing, and presenting clinical documentation) serves four purposes: recording of medical care and communication among providers; payment for hospital and physician; legal defense from medical negligence allegations; and symptom/disease surveillance, public health, and research functions. In the consensus development process described by Handler, these objectives were balanced with the consideration of efficiency, often evaluated as physician time and clinical documentation system costs, in recording the information necessary for their accomplishment. The consensus panel session participants and authors recommend that 1) clinical documentation be electronically retrievable; 2) selection and implementation be evidence-based and grounded on valid metrics (research is needed to identify these metrics); 3) the user interface be crafted to promote clinical excellence through high-quality information collection and efficient charting techniques; 4) the priorities for integration of clinical information be standardized and implemented within enterprises and across health and information systems; 5) systems use accepted standards for bidirectional, real-time clinical data exchange, without limiting the location or number of simultaneous users; 6) systems fully utilize existing electronic sources of specific patient information and general medical knowledge; 7) systems automatically and reliably capture appropriate data that support electronic billing for emergency department services; and 8) systems promote bedside documentation and mobile access. PMID- 15528576 TI - Computerized physician order entry and online decision support. AB - Computerized physician order entry (CPOE) and decision support systems (DSS) can reduce certain types of error but often slow clinicians and may increase other types of error. The net effect of these systems on an emergency department (ED) is unknown. The consensus participants combined published evidence with expert opinion to outline recommendations for success. These include seamless integration of CPOE and DSS into systems and workflow; ensuring access to Internet-based and other online support material in the clinical arena; designing systems specifically for the ED and measuring their impact to ensure an overall benefit; ensuring that CPOE systems provide error and interaction checking and facilitate weight- and physiology-based dosing; using interruptive alerts only for the highest-severity events; providing a simple, vendor-independent interface for institutional customization of CPOE alert thresholds; maximizing the use of automated systems and passive data capture; and ensuring the widespread availability of CPOE and DSS using secure wireless and portable technologies where appropriate. Decisions regarding CPOE and DSS in the ED should be guided by the ED chair or designee. Much of what is believed to be true regarding CPOE and DSS has not been adequately studied. Additional CPOE and DSS research is needed quickly, and this research should receive funding priority. DSS and CPOE hold great promise to improve patient care, but not all systems are equal. Evidence must guide these efforts, and the measured outcomes must consider the many factors of quality care. PMID- 15528577 TI - Disparate systems, disparate data: integration, interfaces, and standards in emergency medicine information technology. AB - As part of the broader informatics consensus initiative sponsored by Academic Emergency Medicine, this report addresses the issues of integration, interfaces, and data standards and how they are relevant to information management in emergency medicine. The purpose of this report, and the workgroup that contributed to its content, is to provide emergency physicians and other stakeholders in the emergency informatics community a sense of direction as they design, build, and/or choose systems. Problems are identified, strategies to address these problems are discussed, and consensus recommendations are provided. PMID- 15528578 TI - See one, do one, teach one: advanced technology in medical education. AB - The concept of "learning by doing" has become less acceptable, particularly when invasive procedures and high-risk care are required. Restrictions on medical educators have prompted them to seek alternative methods to teach medical knowledge and gain procedural experience. Fortunately, the last decade has seen an explosion of the number of tools available to enhance medical education: web based education, virtual reality, and high fidelity patient simulation. This paper presents some of the consensus statements in regard to these tools agreed upon by members of the Educational Technology Section of the 2004 AEM Consensus Conference for Informatics and Technology in Emergency Department Health Care, held in Orlando, Florida. FINDINGS: Web-based teaching: 1) Every ED should have access to medical educational materials via the Internet, computer-based training, and other effective education methods for point-of-service information, continuing medical education, and training. 2) Real-time automated tools should be integrated into Emergency Department Information Systems [EDIS] for contemporaneous education. Virtual reality [VR]: 1) Emergency physicians and emergency medicine societies should become more involved in VR development and assessment. 2) Nationally accepted protocols for the proper assessment of VR applications should be adopted and large multi-center groups should be formed to perform these studies. High-fidelity simulation: Emergency medicine residency programs should consider the use of high-fidelity patient simulators to enhance the teaching and evaluation of core competencies among trainees. CONCLUSIONS: Across specialties, patient simulation, virtual reality, and the Web will soon enable medical students and residents to... see one, simulate many, do one competently, and teach everyone. PMID- 15528579 TI - Information technology principles for management, reporting, and research. AB - Information technology holds the promise to enhance the ability of individuals and organizations to manage emergency departments, improve data sharing and reporting, and facilitate research. The Society for Academic Emergency Medicine (SAEM) Consensus Committee has identified nine principles to outline a path of optimal features and designs for current and future information technology systems. The principles roughly summarized include the following: utilize open database standards with clear data dictionaries, provide administrative access to necessary data, appoint and recognize individuals with emergency department informatics expertise, allow automated alert and proper identification for enrollment of cases into research, provide visual and statistical tools and training to analyze data, embed automated configurable alarm functionality for clinical and nonclinical systems, allow multiexport standard and format configurable reporting, strategically acquire mission-critical equipment that is networked and capable of automated feedback regarding functional status and location, and dedicate resources toward informatics research and development. The SAEM Consensus Committee concludes that the diligent application of these principles will enhance emergency department management, reporting, and research and ultimately improve the quality of delivered health care. PMID- 15528581 TI - Evaluation of emergency medical text processor, a system for cleaning chief complaint text data. AB - OBJECTIVES: Emergency Medical Text Processor (EMT-P) version 1, a natural language processing system that cleans emergency department text (e.g., chst pn, chest pai), was developed to maximize extraction of standard terms (e.g., chest pain). The authors compared the number of standard terms extracted from raw chief complaint (CC) data with that for CC data cleaned with EMT-P and evaluated the accuracy of EMT-P. METHODS: This cross-sectional observation study included CC text entries for all emergency department visits to three tertiary care centers in 2001. Terms were extracted from CC entries before and after cleaning with EMT P. Descriptive statistics included number and percentage of all entries (tokens) and all unique entries (types) that matched a standard term from the Unified Medical Language System (UMLS). An expert panel rated the accuracy of the CC-UMLS term matches; inter-rater reliability was measured with kappa. RESULTS: The authors collected 203,509 CC entry tokens, of which 63,946 were unique entry types. For the raw data, 89,337 tokens (44%) and 5,081 types (8%) matched a standard term. After EMT-P cleaning, 168,050 tokens (83%) and 44,430 types (69%) matched a standard term. The expert panel reached consensus on 201 of the 222 CC UMLS term matches reviewed (kappa=0.69-0.72). Ninety-six percent of the 201 matches were rated equivalent or related. Thirty-eight percent of the nonmatches were found to match UMLS concepts. CONCLUSIONS: EMT-P version 1 is relatively accurate, and cleaning with EMT-P improved the CC-UMLS term match rate over raw data. The authors identified areas for improvement in future EMT-P versions and issues to be resolved in developing a standard CC terminology. PMID- 15528580 TI - Clinical information systems: instant ubiquitous clinical data for error reduction and improved clinical outcomes. AB - Immediate access to existing clinical information is inadequate in current medical practice; lack of existing information causes or contributes to many classes of medical error, including diagnostic and treatment error. A review of the literature finds ample evidence to support a description of the problems caused by data that are missing or unavailable but little evidence to support one proposed solution over another. A primary recommendation of the Consensus Committee is that hospitals and departments should adopt systems that provide fast, ubiquitous, and unified access to all types of existing data. Additional recommendations cover a variety of related functions and operational concepts, from backups and biosurveillance to speed, training, and usability. PMID- 15528582 TI - Discrete event simulation of emergency department activity: a platform for system level operations research. AB - OBJECTIVES: This article explores the potential of discrete event simulation (DES) methods to advance system-level investigation of emergency department (ED) operations. To this end, the authors describe the development and operation of Emergency Department SIMulation (EDSIM), a new platform for computer simulation of ED activity at a Level 1 trauma center. The authors also demonstrate one potential application of EDSIM by using simulated ED activity to compare two patient triage methods. METHODS: The Extend DES modeling package was used to develop a model of ED activity for a five-day period in July 2003. Model input includes staffing levels, facility characteristics, and patient data drawn from electronic patient tracking databases, billing records, and a detailed review of 674 ED charts. The accuracy of model output was tested by comparing predicted and known patient service times. The EDSIM model was then used to compare the fast track triage approach with an alternative acuity ratio triage (ART) approach whereby patients were assigned to staff on an acuity ratio basis. RESULTS: The EDSIM model predicts average patient service times within 10% of actual values. The accuracy of individual patient paths, however, was variable. In the authors' model, 28% of individual patient treatment times had an absolute error of less than one hour, and 59% less than three hours. A preliminary comparison of two triage methods showed that the ART approach reduced imaging bottlenecks and average treatment times for high-acuity patients, but resulted in an overall increase in average service time for low-acuity patients. CONCLUSIONS: The EDSIM model provides a flexible platform for studying ED operations as they relate to average treatment times for ED patients, but the model will require further refinement to predict individual patient times. A comparative study of triage methods suggests that ART provides a mix of benefits and drawbacks, but further investigation will be required to substantiate these preliminary findings. PMID- 15528583 TI - Supporting clinical practice at the bedside using wireless technology. AB - OBJECTIVES: Despite studies that show improvements in both standards of care and outcomes with the judicious application of clinical practice guidelines (CPGs), their clinical utilization remains low. This randomized controlled trial examined the use of a wirelessly networked mobile computer (MC) by physicians at the bedside with access to an emergency department information system, decision support tools (DSTs), and other software options. METHODS: Each of ten volunteer emergency physicians was randomized using a matched-pair design to work five shifts in standard fashion (desktop computer [DC] access) and five shifts with a wirelessly networked MC. Work pattern issues and electronic CPG/DST use were compared using end-of-shift satisfaction questionnaires and review of a CPG/DST database. Repeated-measures analysis of variance was used to examine between shift differences. RESULTS: A total of 100 eight-hour shifts were evaluated; 99% compliance with postshift questionnaires was achieved. Using a seven-point Likert scale (MC values first), MCs were rated as being as fast (5.04 vs. 4.54; p=0.13) and convenient (5.08 vs. 4.14; p=0.07) as DCs. Overall, physicians rated MCs to be less efficient (3.18 vs. 4.30; p=0.02) but encouraged more frequent use of DSTs (4.10 vs. 3.47; p=0.03) without impacting doctor-patient communication (2.78 vs. 2.96; p=0.51). During the study period, physician use of an intranet Web application (eCPG) was more frequent during shifts assigned to the MC when compared with the DC (eCPG uses/shift, 3.6 vs. 2.0; p=0.033). CONCLUSIONS: The MC technology permitted physicians to access information at the bedside and increased the use of CPG/DST tools. According to physicians, patients appeared to accept their use of information technology to assist in decision making. Development of improved computer technology may address the major limitation of MC portability. PMID- 15528584 TI - Virtual TeleStroke support for the emergency department evaluation of acute stroke. AB - OBJECTIVES: Telemedicine-enabled acute stroke consultation (TeleStroke) may be useful to determine eligibility for treatment with tissue plasminogen activator (tPA) and provide support to emergency departments without on-site stroke expertise. METHODS: Emergency physicians consulted with stroke neurologists via two-way videoconferencing in the evaluation of patients with possible acute stroke. History, neurologic examination, and computed tomography of the head were reviewed to determine eligibility for treatment with tPA. Interpretations of computed tomography were compared for inter-rater reliability between the neurologist and the neuroradiologist using a conventional workstation. Videotape and written records were analyzed to determine time intervals, patient management, and user satisfaction. RESULTS: The authors reviewed data from 24 patients evaluated over 27 months at an island-based hospital. The mean National Institutes of Health Stroke Scale score was 5.7 (range, 0-22). Fifteen patients arrived at the emergency department less than three hours after symptom onset; 12 were presented for TeleStroke consultation within three hours after symptom onset. Eight of these 12 (75%) had acute ischemic stroke, and six of these eight potentially eligible patients (75%) received intravenous tPA. There was very good agreement among all remote readers for detecting the one case of imaging exclusion (subdural hemorrhage). There were no protocol violations, and a mean (+/- SD) consult-to-needle time of 36 (+/- 15) minutes and door-to-needle time of 106 (+/- 22) minutes was achieved. Transfer was avoided in 11 patients. Physicians believed that TeleStroke consultation improved care in >95% of the cases. CONCLUSIONS: TeleStroke videoconferencing can support emergency department based evaluation of acute stroke and may facilitate tPA delivery in neurologically underserved facilities. A prospective, randomized trial is needed to determine if these systems are superior to traditional telephone consultation. PMID- 15528585 TI - Medical informatics standards applicable to emergency department information systems: making sense of the jumble. AB - The adoption of medical informatics standards by emergency department information systems (EDISs) is not universal, despite obvious benefits. Clinicians and administrators looking to obtain an EDIS need to know exactly what the various standards can do for them and how the systems they depend on can be integrated and extended. In addition to the standard methods for systems to communicate (chiefly Health Level 7 [HL7]) and those required for submission of claims (Current Procedural Terminology [CPT]-4, International Classification of Diseases, Ninth Revision, Clinical Modification [ICD-9-CM], and X12N), there are several other available standards that are clinically useful and can greatly improve the ability to access and exchange patient information. Major advances in the Unified Medical Language System of the National Library of Medicine have made the patient medical record information standards (Systematized Nomenclature of Medicine [SNOMED], Logical Observation Identifiers, Names, and Codes [LOINC], RxNorm) easily accessible. Detailed knowledge of the arcana associated with the technical aspects of the standards is not needed (or desired) by clinicians to use standards-based systems. However, some knowledge about the commonly used standards is helpful in choosing an EDIS, interfacing the EDIS with the other hospital information systems, extending or upgrading systems, and adopting decision support technologies. PMID- 15528586 TI - Using online analytical processing to manage emergency department operations. AB - The emergency department (ED) is a unique setting in which to explore and evaluate the utility of information technology to improve health care operations. A potentially useful software tool in managing this complex environment is online analytical processing (OLAP). An OLAP system has the ability to provide managers, providers, and researchers with the necessary information to make decisions quickly and effectively by allowing them to examine patterns and trends in operations and patient flow. OLAP software quickly summarizes and processes data acquired from a variety of data sources, including computerized ED tracking systems. It allows the user to form a comprehensive picture of the ED from both system-wide and patient-specific perspectives and to interactively view the data using an approach that meets his or her needs. This article describes OLAP software tools and provides examples of potential OLAP applications for care improvement projects, primarily from the perspective of the ED. While OLAP is clearly a helpful tool in the ED, it is far more useful when integrated into the larger continuum of health information systems across a hospital or health care delivery system. PMID- 15528587 TI - Emergency department digital radiology: moving from photos to pixels. AB - Emergency department (ED) patient care relies heavily on radiologic imaging. As advances in technologic innovation continue to present opportunities to streamline and simplify the delivery of care, emergency medicine (EM) practitioners face the challenge of transitioning from a system of primarily film based radiography to one that utilizes digitized images. The move to digital radiology can result in enhanced quality of patient care, reduction of errors, and increased ED efficiency; however, making this transition will necessarily involve changes in EM practice. As the technology evolves, digital radiology will gradually become ingrained into everyday practice because of these and other notable benefits; however, EM practitioners will need to overcome several challenges to make the transition smoothly and consider the potential impacts that this change will have on ED workflow. The authors discuss the benefits, challenges, and other operational considerations involved with the ED implementation of digital radiology and close by presenting guiding principles for current and future users. Despite the unresolved issues, digital radiology will mature as a technology and improve EM practice, making it one of the great information technology advances in EM. PMID- 15528588 TI - Automating research data collection. AB - This article reviews the capabilities, advantages, and disadvantages of three forms of automated data collection-scannable data forms, Web-based forms, and handheld computers-compared with the current standard of data entry by hand on paper forms. Each of these methods is reviewed with respect to ease of use, experience required of designer, end-user training requirements, costs, flexibility, speed, accuracy/error rate, potential for data loss, need for technical support, and equipment and/or software requirements. A discussion of their appropriate application to various kinds of studies is included, followed by examples of research studies using each of these methods. PMID- 15528589 TI - Information technology and emergency medical care during disasters. AB - Disaster response to mass-casualty incidents represents one of the greatest challenges to a community's emergency response system. Rescuers, field medical personnel, and regional emergency departments and hospitals must often provide care to large numbers of casualties in a setting of limited resources, inadequate communication, misinformation, damaged infrastructure, and great personal risk. Emergency care providers and incident managers attempt to procure and coordinate resources and personnel, often with inaccurate data regarding the true nature of the incident, needs, and ongoing response. In this chaotic environment, new technologies in communications, the Internet, computer miniaturization, and advanced "smart devices" have the potential to vastly improve the emergency medical response to such mass-casualty incident disasters. In particular, next generation wireless Internet and geopositioning technologies may have the greatest impact on improving communications, information management, and overall disaster response and emergency medical care. These technologies have applications in terms of enhancing mass-casualty field care, provider safety, field incident command, resource management, informatics support, and regional emergency department and hospital care of disaster victims. PMID- 15528590 TI - Using data from hospital information systems to improve emergency department care. AB - The ubiquity of computerized hospital information systems, and of inexpensive computing power, has led to an unprecedented opportunity to use electronic data for quality improvement projects and for research. Although hospitals and emergency departments vary widely in their degree of integration of information technology into clinical operations, most have computer systems that manage emergency department registration, admission-discharge-transfer information, billing, and laboratory and radiology data. These systems are designed for specific tasks, but contain a wealth of detail that can be used to educate staff and improve the quality of care emergency physicians offer their patients. In this article, the authors describe five such projects that they have performed and use these examples as a basis for discussion of some of the methods and logistical challenges of undertaking such projects. PMID- 15528591 TI - A real-time tracking, notification, and web-based enrollment system for emergency department research. AB - The authors describe the development of a real-time tracking, notification, and Web-based enrollment system designed specifically to facilitate emergency department research. The system was developed in a cooperative arrangement between an emergency medicine researcher and a medical information software company. The system design and utilization are described as well as the security measures to ensure compliance with Health Insurance Portability and Accountability Act (HIPAA) regulations and database security. PMID- 15528592 TI - Betting your life on it. PMID- 15528593 TI - Developing primary palliative care. PMID- 15528594 TI - Emergency care networks. PMID- 15528595 TI - Long term outcome of treating schizophrenia. PMID- 15528596 TI - Hepatitis B infections. PMID- 15528597 TI - England to start national bowel cancer screening programme. PMID- 15528598 TI - UK clinic allowed to screen embryos for rare bowel cancer. PMID- 15528599 TI - Australian army faces legal action over mefloquine. PMID- 15528601 TI - Surgeons warn changes will fail without investment in training. PMID- 15528600 TI - US judge halts compulsory anthrax vaccination for soldiers. PMID- 15528602 TI - Committee calls for substantial investment in allergy services. PMID- 15528603 TI - Global spending on health research still skewed towards wealthy nations. PMID- 15528607 TI - More than a fifth of children in Darfur is malnourished. PMID- 15528608 TI - FDA approves implantable chip to access medical records. PMID- 15528610 TI - Bush's plan to screen for mental health meets opposition in Illinois. PMID- 15528612 TI - Doctors who offered "magic potions" found guilty of serious professional misconduct. PMID- 15528613 TI - Numbers starting treatment for drug misuse increase by 20% over two years. PMID- 15528615 TI - BMA calls for continuted restrictions on use of IVF. PMID- 15528619 TI - QT interval increased after single dose of lofexidine. PMID- 15528620 TI - Preventing and treating hepatitis B infection. PMID- 15528621 TI - Care in the early newborn period. PMID- 15528622 TI - Should interventions to reduce respirable pollutants be linked to tuberculosis control programmes? PMID- 15528623 TI - Users' guide to detecting misleading claims in clinical research reports. PMID- 15528624 TI - Monitoring global health: time for new solutions. PMID- 15528632 TI - Recent developments in Bell's palsy: trial for Bell's palsy is in progress in Scotland. PMID- 15528633 TI - Recent developments in Bell's palsy: who should provide the care? PMID- 15528634 TI - Recent developments in Bell's palsy: does a more recent single research paper trump a systematic review? PMID- 15528635 TI - Medical education should include human rights component. PMID- 15528638 TI - Centromere parC of plasmid R1 is curved. AB - The centromere sequence parC of Escherichia coli low-copy-number plasmid R1 consists of two sets of 11 bp iterated sequences. Here we analysed the intrinsic sequence-directed curvature of parC by its migration anomaly in polyacrylamide gels. The 159 bp long parC is strongly curved with anomaly values (k-factors) close to 2. The properties of the parC curvature agree with those of other curved DNA sequences. parC contains two regions of 5-fold repeated iterons separated by 39 bp. We modified 4 bp within this intermediate sequence so that we could analyse the two 5-fold repeated regions independently. The analysis shows that the two repeat regions are not independently curved parts of parC but that the overall curvature is a property of the whole fragment. Since the centromere sequence of an E.coli plasmid as well as eukaryotic centromere sequences show DNA curvature, we speculate that curvature might be a general property of centromeres. PMID- 15528640 TI - Genome update: DNA repeats in bacterial genomes. PMID- 15528639 TI - Cis-acting regulatory sequences promote high-frequency gene conversion between repeated sequences in mammalian cells. AB - In mammalian cells, little is known about the nature of recombination-prone regions of the genome. Previously, we reported that the immunoglobulin heavy chain (IgH) mu locus behaved as a hotspot for mitotic, intrachromosomal gene conversion (GC) between repeated mu constant (Cmu) regions in mouse hybridoma cells. To investigate whether elements within the mu gene regulatory region were required for hotspot activity, gene targeting was used to delete a 9.1 kb segment encompassing the mu gene promoter (Pmu), enhancer (Emu) and switch region (Smu) from the locus. In these cell lines, GC between the Cmu repeats was significantly reduced, indicating that this 'recombination-enhancing sequence' (RES) is necessary for GC hotspot activity at the IgH locus. Importantly, the RES fragment stimulated GC when appended to the same Cmu repeats integrated at ectopic genomic sites. We also show that deletion of Emu and flanking matrix attachment regions (MARs) from the RES abolishes GC hotspot activity at the IgH locus. However, no stimulation of ectopic GC was observed with the Emu/MARs fragment alone. Finally, we provide evidence that no correlation exists between the level of transcription and GC promoted by the RES. We suggest a model whereby Emu/MARS enhances mitotic GC at the endogenous IgH mu locus by effecting chromatin modifications in adjacent DNA. PMID- 15528641 TI - Policy proposal for publication of papers with data sets from genome-wide studies. PMID- 15528642 TI - An open letter on microarray data from the MGED Society. PMID- 15528643 TI - An unusual integron in Treponema denticola. PMID- 15528644 TI - Nitrate reduction and the nitrogen cycle in archaea. AB - The nitrogen cycle (N-cycle) in the biosphere, mainly driven by prokaryotes, involves different reductive or oxidative reactions used either for assimilatory purposes or in respiratory processes for energy conservation. As the N-cycle has important agricultural and environmental implications, bacterial nitrogen metabolism has become a major research topic in recent years. Archaea are able to perform different reductive pathways of the N-cycle, including both assimilatory processes, such as nitrate assimilation and N(2) fixation, and dissimilatory reactions, such as nitrate respiration and denitrification. However, nitrogen metabolism is much less known in archaea than in bacteria. The availability of the complete genome sequences of several members of the eury- and crenarchaeota has enabled new approaches to the understanding of archaeal physiology and biochemistry, including metabolic reactions involving nitrogen compounds. Comparative studies reveal that significant differences exist in the structure and regulation of some enzymes involved in nitrogen metabolism in archaea, giving rise to important conclusions and new perspectives regarding the evolution, function and physiological relevance of the different N-cycle processes. This review discusses the advances that have been made in understanding nitrate reduction and other aspects of the inorganic nitrogen metabolism in archaea. PMID- 15528645 TI - The Serratia gene cluster encoding biosynthesis of the red antibiotic, prodigiosin, shows species- and strain-dependent genome context variation. AB - The prodigiosin biosynthesis gene cluster (pig cluster) from two strains of Serratia (S. marcescens ATCC 274 and Serratia sp. ATCC 39006) has been cloned, sequenced and expressed in heterologous hosts. Sequence analysis of the respective pig clusters revealed 14 ORFs in S. marcescens ATCC 274 and 15 ORFs in Serratia sp. ATCC 39006. In each Serratia species, predicted gene products showed similarity to polyketide synthases (PKSs), non-ribosomal peptide synthases (NRPSs) and the Red proteins of Streptomyces coelicolor A3(2). Comparisons between the two Serratia pig clusters and the red cluster from Str. coelicolor A3(2) revealed some important differences. A modified scheme for the biosynthesis of prodigiosin, based on the pathway recently suggested for the synthesis of undecylprodigiosin, is proposed. The distribution of the pig cluster within several Serratia sp. isolates is demonstrated and the presence of cryptic clusters in some strains shown. The pig cluster of Serratia marcescens ATCC 274 is flanked by cueR and copA homologues and this configuration is demonstrated in several S. marcescens strains, whilst these genes are contiguous in strains lacking the pig cluster. PMID- 15528646 TI - BLR-1 and BLR-2, key regulatory elements of photoconidiation and mycelial growth in Trichoderma atroviride. AB - In fungi, phototropism, the induction of carotenogenesis and reproductive structures, and resetting of the circadian rhythm are controlled by blue light. Trichoderma atroviride, a fungus used in biological control, sporulates in a synchronized manner following a brief pulse of blue light. Due to its apparent simplicity, this response was chosen for pursuing photoreceptor isolation. Two genes were cloned, blue-light regulators 1 and 2 (blr-1 and blr-2), similar to the Neurospora crassa white-collar 1 and 2, respectively. The BLR-1 protein has all the characteristics of a blue-light photoreceptor, whereas the structure of the deduced BLR-2 protein suggests that it interacts with BLR-1 through PAS domains to form a complex. Disruption of the corresponding genes demonstrated that they are essential for blue-light-induced conidiation. blr-1 and blr-2 were also shown to be essential for the light-induced expression of the photolyase encoding gene (phr-1). Mechanical injury of mycelia was found to trigger conidiation of T. atroviride, a response not described previously. This response was not altered in the mutants. A novel effect of both red and blue light on mycelial growth was found involving another light receptor, which is compensated by the BLR proteins. PMID- 15528647 TI - Comparative genomics of the KdgR regulon in Erwinia chrysanthemi 3937 and other gamma-proteobacteria. AB - In the plant-pathogenic enterobacterium Erwinia chrysanthemi, almost all known genes involved in pectin catabolism are controlled by the transcriptional regulator KdgR. In this study, the comparative genomics approach was used to analyse the KdgR regulon in completely sequenced genomes of eight enterobacteria, including Erw. chrysanthemi, and two Vibrio species. Application of a signal recognition procedure complemented by operon structure and protein sequence analysis allowed identification of new candidate genes of the KdgR regulon. Most of these genes were found to be controlled by the cAMP-receptor protein, a global regulator of catabolic genes. At the next step, regulation of these genes in Erw. chrysanthemi was experimentally verified using in vivo transcriptional fusions and an attempt was made to clarify the functional role of the predicted genes in pectin catabolism. Interestingly, it was found that the KdgR protein, previously known as a repressor, positively regulates expression of two new members of the regulon, phosphoenolpyruvate synthase gene ppsA and an adjacent gene, ydiA, of unknown function. Other predicted regulon members, namely chmX, dhfX, gntB, pykF, spiX, sotA, tpfX, yeeO and yjgK, were found to be subject to classical negative regulation by KdgR. Possible roles of newly identified members of the Erw. chrysanthemi KdgR regulon, chmX, dhfX, gntDBMNAC, spiX, tpfX, ydiA, yeeO, ygjV and yjgK, in pectin catabolism are discussed. Finally, complete reconstruction of the KdgR regulons in various gamma-proteobacteria yielded a metabolic map reflecting a globally conserved pathway for the catabolism of pectin and its derivatives with variability in transport and enzymic capabilities among species. In particular, possible non-orthologous substitutes of isomerase KduI and a new oligogalacturonide transporter in the Vibrio species were detected. PMID- 15528648 TI - The complete sequences of plasmids pB2 and pB3 provide evidence for a recent ancestor of the IncP-1beta group without any accessory genes. AB - The nucleotide sequences of the broad-host-range antibiotic resistance plasmids pB2 (61 kb) and pB3 (56 kb), which were isolated from a wastewater treatment plant, were determined and analysed. Both have a nearly identical IncP-1beta backbone, which diverged early from the sequenced IncP-1beta plasmids R751, pB10, pJP4, pADP1 and pUO1. In contrast to the latter plasmids, the pB2 and pB3 backbone does not seem to have undergone any deletions. The complete partition gene parA is located downstream of the mating pair formation (trb) module. A 14.4 kb or 19.0 kb mobile genetic element is present between traC and parA of pB3 and pB2, respectively. This region is typical for insertions in IncP-1beta plasmids, but the insertion site is unique. Both elements differ only by a duplication in pB2 of a tetA(C)-tetR-tnpA(IS26) fragment. The 5 bp target site duplication and the 26 bp inverted repeats flanking the mobile genetic elements are still intact, indicating that the insertion occurred recently. The element consists of three nested transposable elements: (i) a relict of a Tn402-like transposon with a gene for a new class D beta-lactamase (bla(NPS-2)); (ii) within that, another Tn402 like element with a class 1 integron harbouring the gene cassettes cmlA1 for a chloramphenicol efflux protein and aadA2 encoding a streptomycin/spectinomycin adenylyltransferase, and a copy of IS6100; (iii) into the integrase gene intI1 a tetracycline resistance module tetA(C)-tetR flanked by copies of IS26 is inserted. Interestingly, in contrast to all other IncP-1beta plasmids analysed so far, the oriV region between trfA and klcA is not interrupted by accessory genes, and there is no indication that previously inserted accessory genes have subsequently been deleted. The genes kluAB are also missing in that region and should thus be considered acquired genes. These findings, together with the fact that IncP-1beta plasmids acquired accessory elements at various positions in the backbone, suggest that IncP-1beta plasmids without any accessory genes exist in microbial communities. They must occasionally acquire accessory genes by transposition events, resulting in those plasmids that have been found based on selectable phenotypic traits. PMID- 15528649 TI - The complete genomes of Lactobacillus plantarum and Lactobacillus johnsonii reveal extensive differences in chromosome organization and gene content. AB - The first comprehensive comparative analysis of lactobacilli was done by comparing the genomes of Lactobacillus plantarum (3.3 Mb) and Lactobacillus johnsonii (2.0 Mb). L. johnsonii is predominantly found in the gastrointestinal tract, while L. plantarum is also found on plants and plant-derived material, and is used in a variety of industrial fermentations. The L. plantarum and L. johnsonii chromosomes have only 28 regions with conservation of gene order, totalling about 0.75 Mb; these regions are not co-linear, indicating major chromosomal rearrangements. Metabolic reconstruction indicates many differences between L. johnsonii and L. plantarum: numerous enzymes involved in sugar metabolism and in biosynthesis of amino acids, nucleotides, fatty acids and cofactors are lacking in L. johnsonii. Major differences were seen in the number and types of putative extracellular proteins, which are of interest because of their possible role in host-microbe interactions. The differences between L. plantarum and L. johnsonii, both in genome organization and gene content, are exceptionally large for two bacteria of the same genus, emphasizing the difficulty in taxonomic classification of lactobacilli. PMID- 15528650 TI - Antibiotic multiresistance plasmid pRSB101 isolated from a wastewater treatment plant is related to plasmids residing in phytopathogenic bacteria and carries eight different resistance determinants including a multidrug transport system. AB - Ten different antibiotic resistance plasmids conferring high-level erythromycin resistance were isolated from an activated sludge bacterial community of a wastewater treatment plant by applying a transformation-based approach. One of these plasmids, designated pRSB101, mediates resistance to tetracycline, erythromycin, roxythromycin, sulfonamides, cephalosporins, spectinomycin, streptomycin, trimethoprim, nalidixic acid and low concentrations of norfloxacin. Plasmid pRSB101 was completely sequenced and annotated. Its size is 47 829 bp. Conserved synteny exists between the pRSB101 replication/partition (rep/par) module and the pXAC33-replicon from the phytopathogen Xanthomonas axonopodis pv. citri. The second pRSB101 backbone module encodes a three-Mob-protein type mobilization (mob) system with homology to that of IncQ-like plasmids. Plasmid pRSB101 is mobilizable with the help of the IncP-1alpha plasmid RP4 providing transfer functions in trans. A 20 kb resistance region on pRSB101 is located within an integron-containing Tn402-like transposon. The variable region of the class 1 integron carries the genes dhfr1 for a dihydrofolate reductase, aadA2 for a spectinomycin/streptomycin adenylyltransferase and bla(TLA-2) for a so far unknown Ambler class A extended spectrum beta-lactamase. The integron-specific 3' segment (qacEDelta1-sul1-orf5Delta) is connected to a macrolide resistance operon consisting of the genes mph(A) (macrolide 2'-phosphotransferase I), mrx (hydrophobic protein of unknown function) and mphR(A) (regulatory protein). Finally, a putative mobile element with the tetracycline resistance genes tetA (tetracycline efflux pump) and tetR was identified upstream of the Tn402-specific transposase gene tniA. The second 'genetic load' region on pRSB101 harbours four distinct mobile genetic elements, another integron belonging to a new class and footprints of two more transposable elements. A tripartite multidrug (MDR) transporter consisting of an ATP-binding-cassette (ABC)-type ATPase and permease, and an efflux membrane fusion protein (MFP) of the RND-family is encoded between the replication/partition and the mobilization module. Homologues of the macrolide resistance genes mph(A), mrx and mphR(A) were detected on eight other erythromycin resistance-plasmids isolated from activated sludge bacteria. Plasmid pRSB101-like repA amplicons were also obtained from plasmid-DNA preparations of the final effluents of the wastewater treatment plant indicating that pRSB101 like plasmids are released with the final effluents into the environment. PMID- 15528651 TI - Identification of Escherichia coli O157 : H7 genes influencing colonization of the bovine gastrointestinal tract using signature-tagged mutagenesis. AB - Enterohaemorrhagic Escherichia coli (EHEC) cause acute gastroenteritis in humans that may be complicated by life-threatening systemic sequelae. The predominant EHEC serotype affecting humans in the UK and North America is O157 : H7 and infections are frequently associated with contact with ruminant faeces. Strategies to reduce the carriage of EHEC in ruminants are expected to lower the incidence of human EHEC infections; however, the molecular mechanisms underlying persistence of EHEC in ruminants are poorly understood. This paper reports the first comprehensive survey for EHEC factors mediating colonization of the bovine intestines by using signature-tagged transposon mutagenesis. Seventy-nine E. coli O157 : H7 mutants impaired in their ability to colonize calves were isolated and 59 different genes required for intestinal colonization were identified by cloning and sequencing of the transposon insertion sites. Thirteen transposon insertions were clustered in the locus of enterocyte effacement (LEE), which encodes a type III protein secretion system required for the formation of attaching and effacing lesions on intestinal epithelia. A putative structural component of the apparatus (EscN) is essential for intestinal colonization; however, the type III secreted effector protein Map plays only a minor role. Other Type III secretion-associated genes were implicated in colonization of calves by E. coli O157 : H7, including z0990 (ecs0850), which encodes the non-LEE encoded type III secreted effector NleD and the closely related z3023 (ecs2672) and z3026 (ecs2674) genes which encode homologues of Shigella IpaH proteins. We also identified a novel fimbrial locus required for intestinal colonization in calves by E. coli O157 : H7 (z2199-z2206; ecs2114-ecs2107/locus 8) and demonstrated that a mutant harbouring a deletion of the putative major fimbrial subunit gene is rapidly out-competed by the parent strain in co-infection studies. Our data provide valuable new information for the development of intervention strategies. PMID- 15528652 TI - Characterization and transcriptional analysis of hupSLW in Gloeothece sp. ATCC 27152: an uptake hydrogenase from a unicellular cyanobacterium. AB - The structural genes (hupSL) encoding an uptake hydrogenase in the unicellular cyanobacterium Gloeothece sp. ATCC 27152, a strain capable of aerobic N(2) fixation, were identified and sequenced. 3'-RACE experiments uncovered the presence of an additional ORF 184 bp downstream of hupL, showing a high degree of sequence identity with a gene encoding an uptake-hydrogenase-specific endopeptidase (hupW) in other cyanobacteria. In addition, the transcription start point was identified 238 bp upstream of the hupS translational start. RT-PCR experiments revealed that hupW is co-transcribed with the uptake hydrogenase structural genes in Gloeothece sp. ATCC 27152. In addition, Northern hybridizations clearly showed that hupSLW are transcribed under nitrogen fixing conditions, but not in the presence of combined nitrogen. A putative NtcA binding site was identified in the promoter region upstream of hupS, centred at -41.5 bp with respect to the transcription start point. Electrophoretic retardation of a labelled DNA fragment (harbouring the putative NtcA-binding motif) was significantly affected by an Escherichia coli cell-free extract containing overexpressed NtcA, suggesting that NtcA is involved in the transcriptional regulation of hupSLW. PMID- 15528653 TI - The siderophore-mediated iron acquisition systems of Acinetobacter baumannii ATCC 19606 and Vibrio anguillarum 775 are structurally and functionally related. AB - The Acinetobacter baumannii type strain, ATCC 19606, secretes acinetobactin, a catechol siderophore highly related to the iron chelator anguibactin produced by the fish pathogen Vibrio anguillarum (Listonella anguillarum). This paper reports the initial characterization of the genes and gene products involved in the acinetobactin-mediated iron-acquisition process. Insertional mutagenesis resulted in the isolation of several derivatives whose ability to grow in medium containing the iron chelator 2,2'-dipyridyl was affected. One of the insertions disrupted a gene encoding a predicted outer-membrane protein, named BauA, highly similar to FatA, the receptor for ferric anguibactin. Immunological relatedness of BauA with FatA was confirmed by Western blot analysis. Another transposon insertion was mapped to a gene encoding a protein highly similar to FatD, the permease component of the anguibactin transport system. Further DNA sequencing and nucleotide sequence analysis revealed that these A. baumannii 19606 genes are part of a polycistronic locus that contains the bauDCEBA ORFs. While the translation products of bauD, -C, -B and -A are highly related to the V. anguillarum FatDCBA iron-transport proteins, the product of bauE is related to the ATPase component of Gram-positive ATP-binding cassette (ABC) transport systems. This entire locus is flanked by genes encoding predicted proteins related to AngU and AngN, V. anguillarum proteins required for the biosynthesis of anguibactin. These protein similarities, as well as the structural similarity of anguibactin and acinetobactin, suggested that these two siderophores could be utilized by both bacterial strains, a possibility that was confirmed by siderophore utilization bioassays. Taken together, these results demonstrate that these pathogens, which cause serious infections in unrelated hosts, express very similar siderophore-mediated iron-acquisition systems. PMID- 15528654 TI - Autogenous modulation of the Bacillus subtilis sacB-levB-yveA levansucrase operon by the levB transcript. AB - Silencing of levB, the second structural gene of the tricistronic levansucrase operon encoding the endolevanase LevB, decreases the level of levansucrase expression in Bacillus subtilis. Conversely, independent expression of levB greatly stimulates operon expression. This autogenous effect is mediated by the levB transcript, which carries an internal sequence (5'-AAAGCAGGCAA-3') involved in the enhancing effect. In vitro, the levB transcript displays an affinity for the N-terminal fragment of SacY (K(D) 0.2 microM), the regulatory protein that prevents transcription termination of the levansucrase operon. This positive feedback loop leads to an increase in the operon expression when B. subtilis is growing in the presence of high sucrose concentrations. Under these conditions, extracellular levan synthesized by the fructosyl polymerase activity of levansucrase can be degraded mainly into levanbiose by the action of LevB. Levanbiose is neither taken up nor metabolized by the bacteria. This work modifies the present view of the status of levansucrase in B. subtilis physiology. PMID- 15528655 TI - Glucan synthesis in the genus Lactobacillus: isolation and characterization of glucansucrase genes, enzymes and glucan products from six different strains. AB - Members of the genera Streptococcus and Leuconostoc synthesize various alpha glucans (dextran, alternan and mutan). In Lactobacillus, until now, the only glucosyltransferase (GTF) enzyme that has been characterized is gtfA of Lactobacillus reuteri 121, the first GTF enzyme synthesizing a glucan (reuteran) that contains mainly alpha-(1-->4) linkages together with alpha-(1-->6) and alpha (1-->4,6) linkages. Recently, partial sequences of glucansucrase genes were detected in other members of the genus Lactobacillus. This paper reports, for the first time, isolation and characterization of dextransucrase and mutansucrase genes and enzymes from various Lactobacillus species and the characterization of the glucan products synthesized, which mainly have alpha-(1-->6)- and alpha-(1- >3)-glucosidic linkages. The four GTF enzymes characterized from three different Lb. reuteri strains are highly similar at the amino acid level, and consequently their protein structures are very alike. Interestingly, these four Lb. reuteri GTFs have relatively large N-terminal variable regions, containing RDV repeats, and relatively short putative glucan-binding domains with conserved and less conserved YG-repeating units. The three other GTF enzymes, isolated from Lactobacillus sakei, Lactobacillus fermentum and Lactobacillus parabuchneri, contain smaller variable regions and larger putative glucan-binding domains compared to the Lb. reuteri GTF enzymes. PMID- 15528656 TI - A new Bacillus cereus DNA-binding protein, HlyIIR, negatively regulates expression of B. cereus haemolysin II. AB - Haemolysin II, HlyII, is one of several cytotoxic proteins produced by Bacillus cereus, an opportunistic human pathogen that causes food poisoning. The hlyII gene confers haemolytic activity to Escherichia coli cells. Here a new B. cereus gene, hlyIIR, which is located immediately downstream of hlyII and regulates hlyII expression, is reported. The deduced amino acid sequence of HlyIIR is similar to prokaryotic DNA-binding transcriptional regulators of the TetR/AcrA family. Measurements of haemolytic activity levels and of hlyII promoter activity levels using gene fusions and primer-extension assays demonstrated that, in E. coli, hlyII transcription decreased in the presence of hlyIIR. Recombinant HlyIIR binds to a 22 bp inverted DNA repeat centred 48 bp upstream of the hlyII promoter transcription initiation point. In vitro transcription studies showed that HlyIIR inhibits transcription from the hlyII promoter by binding to the 22 bp repeat and RNA polymerase, and by decreasing the formation of the catalytically competent open promoter complex. PMID- 15528657 TI - The C-terminal receiver domain of the Rhizobium leguminosarum bv. viciae FixL protein is required for free-living microaerobic induction of the fnrN promoter. AB - The Rhizobium leguminosarum bv. viciae VF39 FixL protein belongs to a distinct group of hybrid regulatory sensor proteins that bear a covalently linked C terminal receiver domain. FixL has an unorthodox histidine kinase domain, which is shared with many other hybrid regulators. The purified FixL protein had autophosphorylation activity. A truncated protein, lacking the receiver domain, had a much-reduced autophosphorylation activity. However, this truncated protein still efficiently phosphorylated the purified receiver domain in trans. This indicates that, in the full-length FixL protein, the conserved histidine residue in the kinase domain is phosphorylated only transiently and that most of the phosphoryl label accumulates in the C-terminal receiver domain. Gene-fusion studies showed that the fixL gene is required for free-living microaerobic induction of the fnrN promoter. The presence of a functional fixK gene is not required. An R. leguminosarum strain lacking fixL could not be complemented with a truncated copy of the gene lacking the receiver domain. This indicates that the C-terminal receiver domain is an intermediate in the signal transduction pathway that links oxygen limitation to induction of the fnrN promoter in R. leguminosarum bv. viciae VF39. PMID- 15528658 TI - Functional characterization of the BvgAS two-component system of Bordetella holmesii. AB - The BvgAS two-component system is the master regulator of virulence gene expression in the mammalian pathogens Bordetella pertussis, Bordetella parapertussis and Bordetella bronchiseptica. This paper reports the partial cloning and characterization of the bvgAS loci of the 'new' Bordetella species Bordetella holmesii, Bordetella trematum and Bordetella hinzii, which are increasingly recognized as opportunistic pathogens in humans. It is demonstrated that the cytoplasmic signalling domains of the BvgS histidine kinases of B. pertussis and B. holmesii are functionally interchangeable, while signal perception by the two sensor proteins seems to be different. Furthermore, it is shown that, despite the high similarity of the BvgA proteins of B. pertussis and B. holmesii, promoter recognition by the response regulator proteins differs substantially in these organisms. PMID- 15528659 TI - A calcium signal is involved in heterocyst differentiation in the cyanobacterium Anabaena sp. PCC7120. AB - The impact of calcium signals in virtually all cells has led to the study of their role in prokaryotic organisms as stress response modulators. Cell differentiation in adverse conditions is a common Ca(2+)-requiring response. Nitrogen starvation induces the differentiation of N(2)-fixing heterocysts in the filamentous cyanobacterium Anabaena sp. PCC7120. This paper reports the use of a recombinant strain of this organism expressing the photoprotein aequorin to monitor the intracellular free-calcium concentration during the course of heterocyst differentiation. A specific calcium signature that is triggered exclusively when cells are deprived of combined nitrogen and generated by intracellular calcium stores was identified. The intracellular calcium signal was manipulated by treatment with specific calcium drugs, and the effect of such manipulation on the process of heterocyst differentiation was subsequently assessed. Suppression, magnification or poor regulation of this signal prevented the process of heterocyst differentiation, thereby suggesting that a calcium signal with a defined set of kinetic parameters may be required for differentiation. A hetR mutant of Anabaena sp. PCC7120 that cannot differentiate into heterocysts retains, however, the capacity to generate the calcium transient in response to nitrogen deprivation, strongly suggesting that Ca(2+) may be involved in a very early step of the differentiation process. PMID- 15528660 TI - Identification and occurrence of tetrad-forming Alphaproteobacteria in anaerobic aerobic activated sludge processes. AB - In an acetate-fed anaerobic-aerobic membrane bioreactor, a deteriorated enhanced biological phosphorus removal (EBPR) community was developed (as determined based on the chemical profiles of organic substrate, soluble phosphate, and intracellular carbohydrate and polyhydroxyalkanote (PHA) concentrations). Microscopic observations revealed the dominance of tetrad-forming organisms (TFOs), of which the majority stained positively for PHA under anaerobic conditions. Fluorescence in situ hybridization (FISH) confirmed that the Alphaproteobacteria (85.0+/-7.0% of total cells) were the most dominant group. A 16S rRNA gene clone library specific for the Alphaproteobacteria indicated that most 16S rRNA gene clones (61% of total clones) were closely affiliated with 'Defluvicoccus vanus', forming a cluster within subgroup 1 of the Alphaproteobacteria. Combined PHA staining and FISH with specific probes designed for the members of the 'Defluvicoccus' cluster suggested diversity within this TFO cluster, and that these TFOs were newly identified glycogen-accumulating organisms in EBPR systems. However, these 'Defluvicoccus'-related TFOs were only seen in low abundance in 12 different EBPR and non-EBPR systems, suggesting that they were not the key populations responsible for the deterioration of full-scale EBPR processes. PMID- 15528661 TI - Novel organization of genes in a phthalate degradation operon of Mycobacterium vanbaalenii PYR-1. AB - Mycobacterium vanbaalenii PYR-1 is capable of degrading polycyclic aromatic hydrocarbons (PAHs) to ring cleavage metabolites. This study identified and characterized a putative phthalate degradation operon in the M. vanbaalenii PYR-1 genome. A putative regulatory protein (phtR) was encoded divergently with five tandem genes: phthalate dioxygenase large subunit (phtAa), small subunit (phtAb), phthalate dihydrodiol dehydrogenase (phtB), phthalate dioxygenase ferredoxin subunit (phtAc) and phthalate dioxygenase ferredoxin reductase (phtAd). A 6.7 kb EcoRI fragment containing these genes was cloned into Escherichia coli and converted phthalate to 3,4-dihydroxyphthalate. Homologues to the operon region were detected in a number of PAH-degrading Mycobacterium spp. isolated from various geographical locations. The operon differs from those of other Gram positive bacteria in both the placement and orientation of the regulatory gene. In addition, the M. vanbaalenii PYR-1 pht operon contains no decarboxylase gene and none was identified within a 37 kb region containing the operon. This study is the first report of a phthalate degradation operon in Mycobacterium spp. PMID- 15528662 TI - A virulence-associated gene microarray: a tool for investigation of the evolution and pathogenic potential of Staphylococcus aureus. AB - An oligonucleotide probe microarray for investigation of the evolution of epidemic Staphylococcus aureus strains has been constructed. The array comprises 383 probes based on virulence-associated genes present in four key strains. Twelve strains including seven for which the complete chromosomal nucleotide sequence was available were tested on the array. Twenty-six per cent of the probes were able to differentiate between strains to give a minimum of two gene differences between pairs. A gene difference distance tree based on the array data had approximately the same topology as one prepared using concatenated MLST sequences. Differences in the topologies of these trees were found to indicate that large-scale recombination events had occurred during the evolution of the species. One such occurrence appears to have been a key event in the genesis of the EMRSA-15 clone (ST22) that currently represents the most prevalent methicillin-resistant S. aureus (MRSA) in the UK. PMID- 15528663 TI - Characterization of a dam mutant of Haemophilus influenzae Rd. AB - The gene encoding Dam methyltransferase of Haemophilus influenzae was mutagenized by the insertion of a chloramphenicol-resistance cassette into the middle of the Dam coding sequence. This mutant construct was introduced into the H. influenzae chromosome by transformation and selection for Cam(R) transformants. The authors have shown that several phenotypic properties, resistance to antibiotics, dyes and detergent as well as efficiency of transformation, depend on the Dam methylation state of the DNA. Although the major role of the methyl-directed mismatch repair (MMR) system is to repair postreplicative errors, it seems that in H. influenzae its effect is more apparent in repairing DNA damage caused by oxidative compounds. In the dam mutant treated with hydrogen peroxide, MMR is not targeted to newly replicated DNA strands and therefore mismatches are converted into single- and double-strand DNA breaks. This is shown by the increased peroxide sensitivity of the dam mutant and the finding that the sensitivity can be suppressed by a mutH mutation inactivating MMR. In the dam mutant treated with nitrofurazone the resulting damage is not converted into DNA breaks but the high sensitivity is also suppressed by a mutH mutation. PMID- 15528664 TI - Reconstruction of the evolutionary history of the LexA-binding sequence. AB - In recent years, the recognition sequence of the SOS repressor LexA protein has been identified for several bacterial clades, such as the Gram-positive, green non-sulfur bacteria and Cyanobacteria phyla, or the 'Alphaproteobacteria', 'Deltaproteobacteria' and 'Gammaproteobacteria' classes. Nevertheless, the evolutionary relationship among these sequences and the proteins that recognize them has not been analysed. Fibrobacter succinogenes is an anaerobic Gram negative bacterium that branched from a common bacterial ancestor immediately before the Proteobacteria phylum. Taking advantage of its intermediate position in the phylogenetic tree, and in an effort to reconstruct the evolutionary history of LexA-binding sequences, the F. succinogenes lexA gene has been isolated and its product purified to identify its DNA recognition motif through electrophoretic mobility assays and footprinting experiments. After comparing the available LexA DNA-binding sequences with the F. succinogenes one, reported here, directed mutagenesis of the F. succinogenes LexA-binding sequence and phylogenetic analyses of LexA proteins have revealed the existence of two independent evolutionary lanes for the LexA recognition motif that emerged from the Gram-positive box: one generating the Cyanobacteria and 'Alphaproteobacteria' LexA-binding sequences, and the other giving rise to the F. succinogenes and Myxococcus xanthus ones, in a transitional step towards the current 'Gammaproteobacteria' LexA box. The contrast between the results reported here and the phylogenetic data available in the literature suggests that, some time after its emergence as a distinct bacterial class, the 'Alphaproteobacteria' lost its vertically received lexA gene, but received later through lateral gene transfer a new lexA gene belonging to either a cyanobacterium or a bacterial species closely related to this phylum. This constitutes the first report based on experimental evidence of lateral gene transfer in the evolution of a gene governing such a complex regulatory network as the bacterial SOS system. PMID- 15528665 TI - The Pseudomonas aeruginosa global regulator MvaT specifically binds to the ptxS upstream region and enhances ptxS expression. AB - Exotoxin A production in Pseudomonas aeruginosa is regulated positively or negatively by several genes. Two such regulatory genes, ptxR and ptxS, which are divergently transcribed from each other, have been described previously. While computer analysis suggested that the ptxR-ptxS intergenic region contains potential binding sites for several regulatory proteins, the mechanism that regulates the expression of either ptxR or ptxS in P. aeruginosa is not known. The presence of a P. aeruginosa protein complex that specifically binds to a segment within this region was determined. In this study the binding region was localized to a 150 bp fragment of the intergenic region and the proteins that constitute the binding complex were characterized as P. aeruginosa HU and MvaT. Recombinant MvaT was purified as a fusion protein (MAL-MvaT) and shown to specifically bind to the ptxR-ptxS intergenic region. A PAO1 isogenic mutant defective in mvaT, PAODeltamvaT, was constructed and characterized. The lysate of PAODeltamvaT failed to bind to the 150 bp probe. The effect of mvaT on ptxS and ptxR expression was examined using real-time PCR experiments. The expression of ptxS was lower in PAODeltamvaT than in PAO1, but no difference was detected in ptxR expression. These results suggest that MvaT positively regulates ptxS expression by binding specifically to the ptxS upstream region. PMID- 15528666 TI - Protein A gene expression is regulated by DNA supercoiling which is modified by the ArlS-ArlR two-component system of Staphylococcus aureus. AB - Bacterial pathogens such as Staphylococcus aureus undergo major physiological changes when they infect their hosts, requiring the coordinated regulation of gene expression in response to the stresses encountered. Several environmental factors modify the expression of S. aureus virulence genes. This report shows that the expression of spa (virulence gene encoding the cell-wall-associated protein A) is down-regulated by high osmolarity (1 M NaCl, 1 M KCl or 1 M sucrose) in the wild-type strain and upregulated by novobiocin (a DNA gyrase inhibitor that relaxes DNA). A gyrB142 allele corresponding to a double mutation in the B subunit of DNA gyrase relaxed DNA and consequently induced spa expression, confirming that spa expression is regulated by DNA topology. Furthermore, in the presence of novobiocin plus 1 M NaCl, a good correlation was observed between DNA supercoiling and spa expression. The ArlS-ArlR two-component system is involved in the expression of virulence genes such as spa. Presence of an arlRS deletion decreased the effect of DNA supercoiling modulators on spa expression, suggesting that active Arl proteins are necessary for the full effect of DNA gyrase inhibitors and high osmolarity on spa expression. Indeed, evidence is provided for a relationship between the arlRS deletion and topological changes in plasmid DNA. PMID- 15528667 TI - Comparative proteome analysis of Mycobacterium tuberculosis grown under aerobic and anaerobic conditions. AB - Data are presented from two-dimensional (2-D) PAGE analysis of Mycobacterium tuberculosis strain Harlingen grown during aerobic and anaerobic culture, according to a modified Wayne dormancy model. M. tuberculosis cultures were grown to the transition point between exponential growth and stationary phase in the presence of oxygen (7 days) and then part of the cultures was shifted to anaerobic conditions for 16 days. Growth declined similarly during aerobic and anaerobic conditions, whereas the ATP consumption rapidly decreased in the anaerobic cultures. 2-D PAGE revealed 50 protein spots that were either unique to, or more abundant during, anaerobic conditions and 16 of these were identified by MALDI-TOF. These proteins were the alpha-crystalline homologue (HspX), elongation factor Tu (Tuf), GroEL2, succinyl-CoA : 3-oxoacid-CoA transferase (ScoB), mycolic acid synthase (CmaA2), thioredoxin (TrxB2), beta-ketoacyl-ACP synthase (KasB), l-alanine dehydrogenase (Ald), Rv2005c, Rv2629, Rv0560c, Rv2185c and Rv3866. Some protein spots were found to be proteolytic fragments, e.g. HspX and GroEL2. These data suggest that M. tuberculosis induces expression of about 1 % of its genes in response to dormancy. PMID- 15528668 TI - Characterization of the fibrinogen-binding surface protein Fbl of Staphylococcus lugdunensis. AB - The fbl gene of Staphylococcus lugdunensis encodes a protein Fbl that is 58 % identical to the clumping factor A (ClfA) of Staphylococcus aureus. The fbl gene was present in eight clinical isolates of S. lugdunensis. When Fbl was expressed on the surface of Lactococcus lactis it promoted adherence to immobilized fibrinogen and cell clumping in a fibrinogen solution. Purified recombinant Fbl region A bound to immobilized fibrinogen in a dose-dependent manner and inhibited the adherence of both Fbl-expressing and ClfA-expressing strains of L. lactis to fibrinogen. Adherence of S. lugdunensis and L. lactis Fbl(+) to immobilized fibrinogen was also inhibited by rabbit anti-Fbl region A antibodies and rabbit anti-ClfA region A antibodies, as well as by human immunoglobulin with a high level of anti-ClfA antibodies. Alignment of the A domains of CflA and Fbl revealed that all of the ClfA residues implicated in binding to the gamma-chain of fibrinogen are conserved in Fbl. Nevertheless Fbl had a tenfold lower affinity for fibrinogen, suggesting that sequence differences that occur elsewhere in the protein, possibly in beta-strand E of domain N2, affect ligand binding. PMID- 15528669 TI - sigmaB-dependent gene induction and expression in Listeria monocytogenes during osmotic and acid stress conditions simulating the intestinal environment. AB - Listeria monocytogenes must overcome a variety of stress conditions in the host digestive tract to cause foodborne infections. The alternative sigma factor sigma(B), encoded by sigB, is responsible for regulating transcription of several L. monocytogenes virulence and stress-response genes, including genes that contribute to establishment of gastrointestinal infections. A quantitative RT-PCR assay was used to measure mRNA transcript accumulation for the virulence genes inlA and bsh, the stress-response genes opuCA and lmo0669 (encoding a carnitine transporter and an oxidoreductase, respectively) and the housekeeping gene rpoB. Assays were conducted on mid-exponential phase L. monocytogenes cells exposed to conditions reflecting osmotic (0.3 M NaCl) or acid (pH 4.5) conditions typical for the human intestinal lumen. In exponential-phase cells, as well as under osmotic and acid stress, inlA, opuCA and bsh showed significantly lower absolute expression levels in a L. monocytogenes DeltasigB null mutant compared to wild type. A statistical model that normalized target gene expression relative to rpoB showed that accumulation of inlA, opuCA and bsh transcripts was significantly increased in the wild-type strain within 5 min of acid and osmotic stress exposure; lmo0669 transcript accumulation increased significantly only after acid exposure. It was concluded that sigma(B) is essential for rapid induction of the tested stress-response and virulence genes under conditions typically encountered during gastrointestinal passage. As inlA, bsh and opuCA are critical for gastrointestinal infections in animal models, the data also suggest that sigma(B) contributes to the ability of L. monocytogenes to cause foodborne infections. PMID- 15528670 TI - Identification and analysis of a siderophore biosynthetic gene cluster from Agrobacterium tumefaciens C58. AB - Using the complete genome sequence from Agrobacterium tumefaciens C58, the authors identified a secondary metabolite gene cluster that encodes the biosynthesis of a metabolite with siderophore activity. Support for this conclusion came from genetic and regulatory analysis of the gene cluster, along with the purification of a metabolite from A. tumefaciens C58 with iron-chelating activity. Genetic analysis of mutant strains disrupted in this gene cluster showed that these strains grew more slowly than the wild-type strain in medium lacking iron. Additionally, the mutant strains failed to produce a chrome-azurol S-reactive material in liquid or solid medium, and failed to produce the metabolite with iron-chelating characteristics that was identified in the wild type strain. Addition of this purified metabolite to the growth medium of a mutant strain restored its ability to grow in iron-deficient medium. Furthermore, expression of this gene cluster was induced by growth under iron-limiting conditions, suggesting that expression of this gene cluster occurs when iron is scarce. These data are all consistent with the proposal that the proteins encoded by this gene cluster are involved in the production of a siderophore. Interestingly, these proteins show the highest level of amino acid similarity to proteins from a gene cluster found in the filamentous cyanobacterium Nostoc sp. PCC7120, rather than to known siderophore biosynthetic enzymes. Given these properties, it is proposed that the siderophore produced by A. tumefaciens C58 will have a unique chemical structure. Production of the siderophore was not required for virulence of A. tumefaciens when tested with a standard stem inoculation assay. PMID- 15528671 TI - Role of Cal, the colicin A lysis protein, in two steps of colicin A release and in the interaction with colicin A-porin complexes. AB - Release of colicin A was studied in Escherichia coli cells that differed in expressing the colicin A lysis protein (Cal). Pools of released and unreleased colicin A were harvested throughout colicin A induction. The amount of colicin A in each pool varied with the time of induction, allowing the definition of two sequential steps in colicin A release, one of which was dependent on Cal. Each step of colicin A release was differently affected in cells containing Cal mutants in which the N-terminal cysteine residue was substituted by either proline or threonine, preventing them from being acylated and matured. These Cal mutants were only observed in degP cells, indicating that the DegP protease cleaved the unacylated precursor of Cal. Cal was found in the insoluble fraction of the pools of released and unreleased colicin A together with the hetero oligomers of colicin A and porins (colicins Au). The biogenesis of colicins Au was studied in temperature-sensitive secA and secY strains and found to be Sec independent, indicating that they are formed by newly synthesized colicin A binding to mature porins already incorporated in the outer membrane. Cal is a lipoprotein similar to VirB7, a constituent of the type IV secretion system. It would interact with colicins Au to constitute the colicin A export machinery. PMID- 15528672 TI - 2-Methylcitrate-dependent activation of the propionate catabolic operon (prpBCDE) of Salmonella enterica by the PrpR protein. AB - The function of the PrpR protein of Salmonella enterica serovar Typhimurium LT2 was studied in vitro and in vivo. The PrpR protein is a sensor of 2-methylcitrate (2-MC), an intermediate of the 2-methylcitric acid cycle used by this bacterium to convert propionate to pyruvate. PrpR was unresponsive to citrate (a close structural analogue of 2-MC) and to propionate, suggesting that 2-MC, not propionate, is the metabolite that signals the presence of propionate in the environment to S. enterica. prpR alleles encoding mutant proteins with various levels of 2-MC-independent activity were isolated. All lesions causing constitutive PrpR activity were mapped to the N-terminal domain of the protein. Removal of the entire sensing domain resulted in a protein (PrpR(c)) with the highest 2-MC-independent activity. Residue A162 is critical to 2-MC sensing, since the mutant PrpR protein PrpR(A162T) was as active as the PrpR(c) protein in the absence of 2-MC. DNA footprinting studies identified the site in the region between prpR and the prpBCDE operon to which the PrpR protein binds. Analysis of the binding-site sequence revealed two sites with dyad symmetry. Results from DNase I footprinting assays suggested that the PrpR protein may have higher affinity for the site proximal to the P(prpBCDE) promoter. PMID- 15528673 TI - Analysis of Pseudomonas fluorescens F113 genes implicated in flagellar filament synthesis and their role in competitive root colonization. AB - The ability of plant-associated micro-organisms to colonize and compete in the rhizosphere is specially relevant for the biotechnological application of micro organisms as inoculants. Pseudomonads are one of the best root colonizers and they are widely used in plant-pathogen biocontrol and in soil bioremediation. This study analyses the motility mechanism of the well-known biocontrol strain Pseudomonas fluorescens F113. A 6.5 kb region involved in the flagellar filament synthesis, containing the fliC, flaG, fliD, fliS, fliT and fleQ genes and part of the fleS gene, was sequenced and mutants in this region were made. Several non motile mutants affected in the fliC, fliS and fleQ genes, and a fliT mutant with reduced motility properties, were obtained. These mutants were completely displaced from the root tip when competing with the wild-type F113 strain, indicating that the wild-type motility properties are necessary for competitive root colonization. A mutant affected in the flaG gene had longer flagella, but the same motility and colonization properties as the wild-type. However, in rich medium or in the absence of iron limitation, it showed a higher motility, suggesting the possibility of improving competitive root colonization by manipulating the motility processes. PMID- 15528674 TI - Molecular detection of Coxiella burnetii in the sera of patients with Q fever endocarditis or vascular infection. AB - In the absence of a specific diagnosis based on serology, chronic Q fever is inevitably fatal. However, diagnosis is often delayed because the test is not widely available. To shorten the diagnostic delay, we adapted a nested-PCR assay with serum as a template and the LightCycler as a thermal cycler, termed LCN-PCR. We retrospectively and prospectively applied this method to samples from 48 patients diagnosed with Q fever endocarditis or vascular infection and to samples from 100 controls with endocarditis caused by other microorganisms. We also prospectively applied this technique to samples from 30 patients treated for a Q fever endocarditis and to samples from 13 patients with a convalescent acute Q fever with ambiguous immunoglobulin G (IgG) phase I titer. LCN-PCR had a specificity of 100%. It was positive only in samples from patients with evolutive Q fever, as none of the samples from patients with a treated chronic Q fever or with a convalescent acute Q fever presented positive results. When performed prospectively on recently stored sera, the sensitivity of LCN-PCR is 64% (7 of 11 samples; P = 0.004), but the efficiency of LCN-PCR was dramatically altered by the storage of specimens at -20 degrees C. High IgG phase I titers decreased the sensitivity of LCN-PCR. A significant difference was observed among LCN-PCR results for sera with IgG phase I titers of > or =1:25,600 compared to sera with IgG phase I titers of <1:25,600 (0 of 15 samples versus 13 of 33 samples; P = 0.004). In patient samples with titers below 1:25,600 tested prospectively, sensitivity was 100% (7 of 7). The LCN-PCR assay may be helpful in establishing an early diagnosis of chronic Q fever. PMID- 15528675 TI - Development of a PCR method for rapid identification of new Streptococcus mutans serotype k strains. AB - In a previous study, we isolated and characterized a new serotype k of Streptococcus mutans from human blood and oral cavities. Analysis of the genes involved in biosynthesis of the serotype-specific polysaccharide of serotype k strains revealed that the serotype k-specific nucleotide alignment was commonly present in the 5' region of the rgpF gene (350 bp from the initial sequence) compared to the reference strains, and then a method for rapid identification of serotype k strains was developed by use of PCR with primers designed on the basis of the sequence of the variable region. PCR assays with primers specific for amplification of serotype k strains showed a negative reaction with serotype c, e, and f strains and a positive reaction with serotype k strains, with the sensitivity for identification of the serotype k strains shown to range from 5 to 50 cells. Next, the frequency of positive reactions for serotype k-specific primers was surveyed with DNA taken from saliva samples from 200 subjects (2 to 18 years of age), and 10 of those showed a positive reaction, which was higher than the frequency in our previous survey with a serological method. In addition, all saliva samples from subjects with serotype k strains in our previous study were shown to be positive with the serotype k-specific primers. These results indicate that this new PCR method is effective for identification of subjects with S. mutans serotype k. PMID- 15528676 TI - Microsatellite mapping of Mycobacterium leprae populations in infected humans. AB - To investigate genetic diversity in a bacterial population, we measured the copy numbers of simple sequence repeats, or microsatellites, in Mycobacterium leprae from patients living in and around Hyderabad, India. Three microsatellite loci containing trinucleotide or dinucleotide repeats were amplified from infected tissues, and the copy numbers were established by sequence analysis. Extensive diversity was observed in a cross-sectional survey of 33 patients, but closely related profiles were found for members of a multicase family likely to share a common transmission source. Sampling of multiple tissues from single individuals demonstrated identical microsatellite profiles in the skin, nasal cavity, and bloodstream but revealed differences at one or more loci for M. leprae present in nerves. Microsatellite mapping of M. leprae represents a useful tool for tracking short transmission chains. Comparison of skin and nerve lesions suggests that the evolution of disease within an individual involves the expansion of multiple distinct subpopulations of M. leprae. PMID- 15528677 TI - Distribution of putative adhesins in different seropathotypes of Shiga toxin producing Escherichia coli. AB - The distribution of eight putative adhesins that are not encoded in the locus for enterocyte effacement (LEE) in 139 Shiga toxin-producing Escherichia coli (STEC) of different serotypes was investigated by PCR. Five of the adhesins (Iha, Efa1, LPF(O157/OI-141), LPF(O157/OI-154), and LPF(O113)) are encoded in regions corresponding to genomic O islands of E. coli EDL933, while the other three adhesins have been reported to be encoded in the STEC megaplasmid of various serotypes (ToxB [O157:H7], Saa [O113:H21], and Sfp [O157:NM]). STEC strains were isolated from humans (n = 54), animals (n = 52), and food (n = 33). They were classified into five seropathotypes (A through E) based on the reported occurrence of STEC serotypes in human disease, in outbreaks, and in the hemolytic uremic syndrome (M. A. Karmali, M. Mascarenhas, S. Shen, K. Ziebell, S. Johnson, R. Reid-Smith, J. Isaac-Renton, C. Clark, K. Rahn, and J. B. Kaper, J. Clin. Microbiol. 41:4930-4940, 2003). The most prevalent adhesin was that encoded by the iha gene (91%; 127 of 139 strains), which was distributed in all seropathotypes. toxB and efa1 were present mainly in strains of seropathotypes A and B, which were LEE positive. saa was present only in strains of seropathotypes C, D, and E, which were LEE negative. Two fimbrial genes, lpfA(O157/OI-141) and lpfA(O157/OI-154), were strongly associated with seropathotype A. The fimbrial gene lpfA(O113) was present in all seropathotypes except for seropathotype A, while sfpA was not present in any of the strains studied. The distribution of STEC adhesins depends mainly on serotypes and not on the source of isolation. Seropathotype A, which is associated with severe disease and frequently is involved in outbreaks, possesses a unique adhesin profile which is not present in the other seropathotypes. The wide distribution of iha in STEC strains suggested that it could be a candidate for vaccine development. PMID- 15528678 TI - New quadriplex PCR assay for detection of methicillin and mupirocin resistance and simultaneous discrimination of Staphylococcus aureus from coagulase-negative staphylococci. AB - Major challenges in diagnostic molecular microbiology are to develop a simple assay to distinguish Staphylococcus aureus from the less virulent but clinically important coagulase-negative staphylococci (CoNS) and to simultaneously determine their antibiotic resistance profiles. Multiplex PCR assays have been developed for the detection of methicillin- and mupirocin-resistant S. aureus and CoNS but not for the simultaneous discrimination of S. aureus from CoNS. We designed a new set of Staphylococcus genus-specific primers and developed a novel quadriplex PCR assay targeting the 16S rRNA (Staphylococcus genus specific), nuc (S. aureus species specific), mecA (a determinant of methicillin resistance), and mupA (a determinant of mupirocin resistance) genes to identify most staphylococci, to discriminate S. aureus from CoNS and other bacteria, and to simultaneously detect methicillin and mupirocin resistance. Validation of the assay with 96 ATCC control strains and 323 previously characterized clinical isolates, including methicillin- and mupirocin-sensitive and -resistant S. aureus and CoNS isolates and other bacteria, demonstrated 100% sensitivity, specificity, and accuracy. This assay represents a simple, rapid, accurate, and reliable approach for the detection of methicillin- and mupirocin-resistant staphylococci and offers the hope of preventing their widespread dissemination through early and reliable detection. PMID- 15528679 TI - Evaluation of a rapid immunochromatographic assay for identification of Candida albicans and Candida dubliniensis. AB - Candida dubliniensis was first established as a novel yeast species in 1995. It is particularly associated with recurrent episodes of oral candidosis in human immunodeficiency virus (HIV)-infected patients, but it has also been detected at other anatomical sites and at a low incidence level in non-HIV-infected patients. It shares so many phenotypic characteristics with C. albicans that it is easily misidentified as such. No rapid, simple, and commercial test that allows differentiation between C. dubliniensis and C. albicans has been developed, until now. Accurate species identification requires the use of genotype-based techniques that are not routinely available in most clinical microbiology diagnostic laboratories. The present study was designed to evaluate the efficiency of a new test (the immunochromatographic membrane [ICM] albi-dubli test; SR2B, Avrille, France) to differentiate between C. albicans and C. dubliniensis. The organisms evaluated were strains whose identities had previously been confirmed by PCR tests and freshly isolated clinical strains and included 58 C. albicans isolates, 60 C. dubliniensis isolates, and 82 isolates belonging to other species of yeast. The ICM albi-dubli test is based on the principle of immunochromatographic analysis and involves the use of two distinct monoclonal antibodies that recognize two unrelated epitopes expressed by both species or specific to only one species. The assay requires no complex instrumentation for analysis and can be recommended for routine use in clinical microbiology laboratories. Results are obtained within 2 h and 30 min and are easy to interpret. This evaluation demonstrated the good performance of this immunochromatographic test for C. albicans and C. dubliniensis isolated on Sabouraud dextrose agar, CHOROMagar Candida, and CandidaSelect, with sensitivities and specificities ranging from 93.1 to 100%. These parameters decreased, however, to 91.4% when the test was performed with yeast isolated with Candida ID. PMID- 15528680 TI - Characterization of spotted fever group rickettsiae in flea and tick specimens from northern Peru. AB - Evidence of spotted fever group (SFG) rickettsiae was obtained from flea pools and individual ticks collected at three sites in northwestern Peru within the focus of an outbreak of febrile disease in humans attributed, in part, to SFG rickettsia infections. Molecular identification of the etiologic agents from these samples was determined after partial sequencing of the 17-kDa common antigen gene (htrA) as well as pairwise nucleotide sequence homology with one or more of the following genes: gltA, ompA, and ompB. Amplification and sequencing of portions of the htrA and ompA genes in pooled samples (2 of 59) taken from fleas identified the pathogen Rickettsia felis. Four tick samples yielded molecular evidence of SFG rickettsiae. Fragments of the ompA (540-bp) and ompB (2,484-bp) genes were amplified from a single Amblyomma maculatum tick (tick 124) and an Ixodes boliviensis tick (tick 163). The phylogenetic relationships between the rickettsiae in these samples and other rickettsiae were determined after comparison of their ompB sequences by the neighbor-joining method. The dendrograms generated showed that the isolates exhibited close homology (97%) to R. aeschlimannii and R. rhipicephali. Significant bootstrap values supported clustering adjacent to this nodule of the SFG rickettsiae. While the agents identified in the flea and tick samples have not been linked to human cases in the area, these results demonstrate for the first time that at least two SFG rickettsia agents were circulating in northern Peru at the time of the outbreak. Furthermore, molecular analysis of sequences derived from the two separate species of hard ticks identified a possibly novel member of the SFG rickettsiae. PMID- 15528681 TI - Genotypic diversity of Francisella tularensis infecting Dermacentor variabilis ticks on Martha's Vineyard, Massachusetts. AB - Martha's Vineyard, Mass., has been the site of two outbreaks of tularemia (1978 and 2000). Although most patients from both outbreaks presented with pneumonic disease and although aerosol transmission has been suggested, the bite of a dog tick and exposure to rabbits remain the only proven modes of transmission. The factors that precipitated the tularemia outbreaks or the proximal determinants of human risk remain undescribed. We sought to test the hypothesis that the ongoing outbreak is due to a recent introduction event as opposed to amplification of a cryptic enzootic cycle. From 2001-2003, we collected 4,246 dog ticks and tested them in pools for evidence of tularemia by PCR. We then measured the genetic diversity of Francisella tularensis by using multiple-locus variable-number tandem repeat analysis. The prevalence of F. tularensis in dog ticks averaged 0.7%. From 29 positive pools, we identified 10 unique genotypes, which was an unexpectedly large degree of diversity (Simpson's index = 0.86). This degree of genetic diversity is inconsistent with a recent introduction event. We conclude that there has been long-standing enzootic transmission of tularemia on the island. PMID- 15528682 TI - Nasopharyngeal versus oropharyngeal sampling for detection of pneumococcal carriage in adults. AB - Several studies have shown that nasopharyngeal sampling is more sensitive than oropharyngeal sampling for the detection of pneumococcal carriage in children. The data for adults are limited and conflicting. This study was part of a larger study of pneumococcal carriage on the Navajo and White Mountain Apache Reservation following a clinical trial of a seven-valent pneumococcal conjugate vaccine. Persons aged 18 years and older living in households with children enrolled in the vaccine trial were eligible. We collected both nasopharyngeal and oropharyngeal specimens by passing a flexible calcium alginate wire swab either nasally to the posterior nasopharynx or orally to the posterior oropharynx. Swabs were placed in skim milk-tryptone-glucose-glycerin medium and frozen at -70 degrees C. Pneumococcal isolation was performed by standard techniques. Analyses were based on specimens collected from 1,994 adults living in 1,054 households. Nasopharyngeal specimens (11.1%; 95% confidence interval [CI], 9.8 and 12.6%) were significantly more likely to grow pneumococci than were oropharyngeal specimens (5.8%; 95% CI, 4.8 to 6.9%) (P < 0.0001). Few persons had pneumococcal growth from both specimens (1.7%). Therefore, both tests together were more likely to identify pneumococcal carriage (15.2%; 95% CI, 13.7 to 16.9%) than either test alone. Although we found that nasopharyngeal sampling was more sensitive than oropharyngeal sampling, nasopharyngeal sampling alone would have underestimated the prevalence of pneumococcal carriage in this adult population. Sampling both sites may give more accurate results than sampling either site alone in studies of pneumococcal carriage in adults. PMID- 15528683 TI - Evaluation of the etest method using Mueller-Hinton agar with glucose and methylene blue for determining amphotericin B MICs for 4,936 clinical isolates of Candida species. AB - The performance of the Etest using Mueller-Hinton agar supplemented with glucose (2%) and methylene blue (0.5 microg/ml) (MH-GMB) for amphotericin B susceptibility testing of 4,936 isolates of Candida spp. was assessed against that of Etest using RPMI agar with 2% glucose (RPG). MICs were determined by Etest in both media for all 4,936 isolates and were read after incubation for 48 h at 35 degrees C. The Candida isolates included C. albicans (n = 2,728), C. glabrata (n = 722), C. parapsilosis (n = 666), C. tropicalis (n = 528), C. krusei (n = 143), C. lusitaniae (n = 54), C. guilliermondii (n = 39), C. pelliculosa (n = 17), C. kefyr (n = 15), C. rugosa (n = 11), C. dubliniensis (n = 5), C. zeylanoides (n = 4), C. lipolytica (n = 3), and C. famata (n = 1). The Etest results with MH-GMB correlated well with those with RPG. Overall agreement was 92.9%, and agreements for individual species were as follows: C. lusitaniae, 98.1%; C. albicans, 95.1%; C. glabrata, 94.3%; C. krusei, 91.6%; C. parapsilosis, 86.6%; and C. tropicalis, 86.4%. The Etest method using MH-GMB appears to be a useful method for determining amphotericin B susceptibilities of Candida species. PMID- 15528684 TI - Prevalence and molecular analysis of macrolide and fluoroquinolone resistance among isolates of Streptococcus pneumoniae collected during the 2000-2001 PROTEKT US Study. AB - The PROTEKT US (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin in the United States) surveillance program was established to determine the prevalence and mechanisms of antibacterial resistance among bacterial pathogens from patients with community-acquired respiratory tract infections. In year 1 of the PROTEKT US study, 10,103 isolates of Streptococcus pneumoniae, including 3,133 erythromycin-resistant strains and 81 levofloxacin resistant strains, were collected from 206 centers. We report on the molecular analyses of these resistant strains. The resistance genotypes among the 3,044 typed macrolide-resistant isolates overall were mef(A) (n = 2,157; 70.9%), erm(B) (n = 530; 17.4%), mef(A) erm(B) (n = 304; 10.0%), and erm(A) subclass erm(TR) (n = 5; 0.2%). Fifty (1.6%) macrolide-resistant isolates were negative for the mef and the erm resistance genes. Seventy-eight (96.3%) of the 81 levofloxacin resistant isolates analyzed possessed multiple mutations in the gyrA, gyrB, parC, and/or parE quinolone resistance-determining regions. A total of 43 known multilocus sequence typing (MLST) profiles (or single- or double-locus variants) accounted for 75 of 81 isolates. There was no evidence of dissemination of fluoroquinolone-resistant clones within the United States; however, 12 isolates with the same MLST profile were located in one center in Massachusetts. Almost 90% of the erythromycin-resistant isolates and approximately one-third of the levofloxacin-resistant isolates were multidrug resistant. PMID- 15528685 TI - Development and evaluation of a quality-controlled ribosomal sequence database for 16S ribosomal DNA-based identification of Staphylococcus species. AB - To establish an improved ribosomal gene sequence database as part of the Ribosomal Differentiation of Microorganisms (RIDOM) project and to overcome the drawbacks of phenotypic identification systems and publicly accessible sequence databases, both strands of the 5' end of the 16S ribosomal DNA (rDNA) of 81 type and reference strains comprising all validly described staphylococcal (sub)species were sequenced. Assuming a normal distribution for pairwise distances of all unique staphylococcal sequences and choosing a reporting criterion of > or =98.7% similarity for a "distinct species," a statistical error probability of 1.0% was calculated. To evaluate this database, a 16S rDNA fragment (corresponding to Escherichia coli positions 54 to 510) of 55 clinical Staphylococcus isolates (including those of the small-colony variant phenotype) were sequenced and analyzed by the RIDOM approach. Of these isolates, 54 (98.2%) had a similarity score above the proposed threshold using RIDOM; 48 (87.3%) of the sequences gave a perfect match, whereas 83.6% were found by searching National Center for Biotechnology Information (NCBI) database entries. In contrast to RIDOM, which showed four ambiguities at the species level (mainly concerning Staphylococcus intermedius versus Staphylococcus delphini), the NCBI database search yielded 18 taxon-related ambiguities and showed numerous matches exhibiting redundant or unspecified entries. Comparing molecular results with those of biochemical procedures, ID 32 Staph (bioMerieux, Marcy I'Etoile, France) and VITEK 2 (bioMerieux) failed to identify 13 (23.6%) and 19 (34.5%) isolates, respectively, due to incorrect identification and/or categorization below acceptable values. In contrast to phenotypic methods and the NCBI database, the novel high-quality RIDOM sequence database provides excellent identification of staphylococci, including rarely isolated species and phenotypic variants. PMID- 15528686 TI - Evaluation of a multiplexed bead assay for assessment of Epstein-Barr virus immunologic status. AB - Currently, serological assays using either indirect immunofluorescence assay or enzyme-linked immunosorbent assay (ELISA) are performed to evaluate the status of Epstein-Barr virus (EBV) infection in humans. Although these methods are reliable, they are limited to testing an antibody response to a single viral antigen per reaction, thus necessitating a panel of assays to complete the evaluation. In contrast, a new bead-based method (BioPlex 2200; Bio-Rad Laboratories, Hercules, Calif.) can analyze the humoral response to multiple antigens in a single tube. This approach potentially reduces overall cost, turnaround time, and sample volume. The aim of this study was to evaluate the multiplexed EBV serologic assays performed on the BioPlex 2200 platform compared to results of conventional heterophile and ELISA-based assays. A total of 167 nonconsecutive, stored serum samples from adult and pediatric patients submitted for EBV serologic studies were used in the evaluation. Concordance between results generated by the BioPlex 2200 system and conventional assays was calculated. The anti-EA-D assay had the lowest concordance at 91%. The BioPlex 2200 system showed 97% agreement with conventional heterophile and anti-nuclear antigen assays and 92% agreement with the anti-VCA IgG and immunoglobulin M assays. Agreement between the BioPlex 2200 system and conventional testing was 92% with respect to categorization of acute versus nonacute EBV disease. The correlation between these two systems with regard to assignment into one of four categories of EBV status was also good (82%). In summary, there is excellent correlation between contemporary EBV serologic testing and the BioPlex 2200 system. PMID- 15528687 TI - Application of mycobacterial interspersed repetitive unit typing to Manitoba tuberculosis cases: can restriction fragment length polymorphism be forgotten? AB - Since 1993, all Mycobacterium tuberculosis isolates recovered in the province of Manitoba, Canada, have been genotyped by the standard IS6110-restriction fragment length polymorphism (RFLP) method for routine surveillance, prevention, and control purposes. To date, our laboratory has collected 1,290 isolates, from which we have identified approximately 390 unique fingerprint patterns or "types." Although the standard method is well known for being a lengthy and labor intensive procedure, a more efficient alternative for typing tuberculosis isolates, the mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR) method, has recently gained acceptance. Consequently, all isolates acquired in 2003 (n = 126) were typed by both methods in order to determine the utility of replacing the RFLP method with MIRU typing for all future isolates. Application of Hunter's discriminatory index to the available study population showed that the MIRU method was close in discriminatory power (D) to the RFLP method (D(MIRU) = 0.831 to 0.984 versus D(RFLP) = 0.821 to 0.997). Clustering of isolates by using MIRU data correlated with RFLP-derived clustering, lending useful information for either an investigation or confirmation of an incidence of recent transmission. In addition, it was determined that each predominant RFLP type in Manitoba had a corresponding, recognizable MIRU type. It is conceivable that in the future RFLP typing can be replaced with MIRU for real-time, ongoing tuberculosis surveillance in the province. PMID- 15528688 TI - Detection and selection of microsatellites in the genome of Paracoccidioides brasiliensis as molecular markers for clinical and epidemiological studies. AB - Paracoccidioides brasiliensis, a thermodimorphic fungus, is the causative agent of the prevalent systemic mycosis in Latin America, paracoccidioidomycosis (PCM). Here, we describe the microsatellite patterns observed in a collection of P. brasiliensis random sequence tags. We identified 1,117 microsatellite patterns in about 3.8 Mb of unique sequences (0.47% of the total DNA used in the analysis). The majority of these microsatellites (87.5%) are found in noncoding sequences. We used two polymorphic microsatellites located on noncoding and coding sequences, as well as two microsatellites located on introns, as molecular markers to discriminate P. brasiliensis isolates, to look for relationships between the genetic background of the strains and the types of human disease they cause. We did not observe any correlation between the clinical form of human PCM and four simple sequence repeat patterns analyzed. PMID- 15528689 TI - Strong association between genotype F and hepatitis B virus (HBV) e antigen negative variants among HBV-infected argentinean blood donors. AB - A number of reports have indicated an increased risk of cirrhosis and hepatocellular carcinoma in hepatitis B virus (HBV)-infected individuals carrying HBV e antigen (HBeAg)-negative variants. Although distinct core promoter and precore mutations distributed according to geographical locality and viral genotype have been reported, epidemiological data from South America are still scarce. The prevalences of HBV genotypes and core promoter and precore polymorphisms in 75 HBeAg-negative Argentinean blood donors were surveyed. The observed frequencies of HBV genotypes were 64.0% for genotype F, 17.3% each for genotypes A and D, and 1.3% for genotype C. Genotype F strains were widely distributed and significantly more prevalent in the northern region of the country (P < 0.001). An overall high proportion of a stop codon mutation (UAG) at precore codon 28 (66.7%) was observed. Wild-type codon 28 (UGG) was present in 29.3% of the samples, and the remaining 4.0% of samples had mixed variants. The combination of A at nucleotide (nt) 1762 and G at nt 1764 of the core promoter was found in 58.7% of the samples. The variant profiles--T at nt 1762 and A at nt 1764 or A at nt 1762 and A at nt 1764--were detected in 28.0 and 1.3% of the samples, respectively. The observed core promoter polymorphisms could not be related to the ratio of HBeAg to anti-HBeAg antibody, HBV genotype, or precore codon 28 status. Nevertheless, a clear association of genotype F and a precore stop codon mutation was found (P < 0.05). In conclusion, HBV genotype F and mutant codon 28 strains predominated and were strongly associated in a geographically broad Argentinean blood donor population. PMID- 15528690 TI - Use of multilocus variable-number tandem-repeat analysis for typing Mycobacterium avium subsp. paratuberculosis. AB - The etiology of Crohn's disease in humans is largely unknown. Clinical signs of Crohn's disease partly resemble the clinical picture of Johne's disease in ruminants caused by Mycobacterium avium subsp. paratuberculosis. Because of the high prevalence of these bacteria in (products of) ruminants and their remarkable thermostability, concern has been raised about the possible role of these bacteria in the pathogenesis of Crohn's disease. In an attempt to develop a molecular typing method to facilitate meaningful comparative DNA fingerprinting of M. avium subsp. paratuberculosis isolates from the human and animal reservoirs, multilocus variable-number tandem-repeat analysis (MLVA) was explored and compared to IS900 restriction fragment length polymorphism (RFLP) typing. MLVA typing subdivided the most predominant RFLP type, R01, into six subtypes and thus provides a promising molecular subtyping approach to study the diversity of M. avium subsp. paratuberculosis. PMID- 15528691 TI - Molecular characteristics of strains of the cameroon family, the major group of Mycobacterium tuberculosis in a country with a high prevalence of tuberculosis. AB - A preliminary investigation of the genetic biodiversity of Mycobacterium tuberculosis complex strains in Cameroon, a country with a high prevalence of tuberculosis, described a group of closely related M. tuberculosis strains (the Cameroon family) currently responsible for more than 40% of smear-positive pulmonary tuberculosis cases. Here, we used various molecular methods to study the genetic characteristics of this family of strains. Cameroon family M. tuberculosis strains (i) are part of the major genetic group 2 and lack the TbD1 region like other families of epidemic strains, (ii) lack spacers 23, 24, and 25 in their direct repeat (DR) region, (iii) have an identical number of repeats in 8 of 12 variable-number tandem repeats of mycobacterial interspersed repetitive unit (MIRU-VNTR) loci, (iv) have similar IS6110-restriction fragment length polymorphism (RFLP) multiband patterns (10 to 15 copies) with seven common IS6110 bands, (v) do not have an IS6110 element in their DR locus, and (vi) have four IS6110 elements in open reading frames (adenylate cyclase, phospholipase C, moeY, and ATP binding genes). Analysis by spoligotyping, MIRU-VNTR, and IS6110-RFLP typing methods revealed differences not observed in previous studies; polymorphism as assessed by MIRU-VNTR typing was lower than suggested by spoligotyping, and in rare cases, strains with identical IS6110-RFLP patterns had spoligotypes differing by as much as 15 spacers. Our findings confirm the recent expansion of this family in Cameroon and indicate that the interpretation of molecular typing results has to be adapted to the characteristics of the strain population within each setting. The knowledge of this particular genotype, with its large involvement in tuberculosis in Cameroon, allows greater refinement of tuberculosis transmission studies by interpreting data in the context of this geographic area. PMID- 15528692 TI - Longitudinal study of hepatitis activity and viral replication before and after HBeAg seroconversion in chronic hepatitis B patients infected with genotypes B and C. AB - The aims of this study were to compare chronic hepatitis B (CHB) patients with genotypes B and C for the probability of HBeAg seroconversion, hepatitis activity, and viral replication before and after HBeAg seroconversion and to compare the prevalence of core promoter and precore mutations. A total of 180 asymptomatic Chinese patients with CHB were monitored for a median of 53.8 months, and 38 patients with cirrhosis-related complications were studied. Hepatitis B virus (HBV) DNA levels were measured in 16 patients with HBeAg seroconversion at 3 months before, during, and 3 months after HBeAg seroconversion and in all patients at the last follow-up. Hepatitis B genotypes and core promoter and precore mutations were determined. Compared to patients with genotype C (n = 109), patients with subtype Ba (n = 69) had a higher rate of anti-HBe positivity on presentation (P = 0.05). HBeAg-positive patients with subtype Ba had a higher cumulative rate of HBeAg seroconversion than patients with genotype C (P = 0.02). However, there were no differences between the two groups with regard to the median HBV DNA levels before, during, and after HBeAg seroconversion; the probability of having persistently normal or elevated aminotransferase levels; and the median HBV DNA levels and liver biochemistry at the last follow-up. There was no difference in the prevalence of genotypes and core promoter and precore mutations between patients with and without cirrhosis related complications. Though patients with subtype Ba had earlier HBeAg seroconversion, the liver biochemistry, HBV DNA levels in different phases of the disease, and the probability of development of cirrhosis-related complications were the same with genotypes Ba and C. PMID- 15528693 TI - Evaluation of the BD PHOENIX automated microbiology system for detection of methicillin resistance in coagulase-negative staphylococci. AB - The new BD PHOENIX automated microbiology system (Becton Dickinson Diagnostic Systems, Sparks, Md.) is designed for automated rapid antimicrobial susceptibility testing and identification of clinically relevant bacteria. In our study, the accuracy and speed of the BD PHOENIX oxacillin MIC determination for detecting methicillin resistance was evaluated for 200 clinical isolates of coagulase-negative staphylococci (CoNS). Compared to mecA PCR, the BD PHOENIX system detected methicillin resistance with a sensitivity of 99.2%. According to the actual NCCLS oxacillin MIC breakpoint of > or =0.5 microg/ml, the specificity was only 64.9%, attributable to false-positive results in 26 mecA-negative strains, including 16 non-Staphylococcus epidermidis strains. Alternative oxacillin breakpoints of > or =1, > or =2, and > or =4 microg/ml resulted in increased specificities of 83.8, 94.6, and 100% and high sensitivities of 99.2, 99.2, and 96.7%, respectively. Similarly, NCCLS broth microdilution oxacillin MICs exhibited a sensitivity of 100% but a low degree of specificity. However, the previous oxacillin MIC breakpoint of > or =4 microg/ml performed with a sensitivity of 98.4% and a specificity of 98.7%. BD PHOENIX oxacillin MIC results were available after 9 h for 40.5% of the examined CoNS strains and were completed after 17 h. Our results revealed the high reliability of the BD PHOENIX system as a phenotypic method for detection of resistance to oxacillin in mecA positive CoNS. However, for the improvement of specificity, reevaluation of the optimal oxacillin MIC breakpoint for CoNS appears to be necessary. PMID- 15528694 TI - Routes of transmission of swine hepatitis E virus in pigs. AB - Hepatitis E virus (HEV) is believed to be transmitted by the fecal-oral route in pigs. To date, in experiments, HEV has been transmitted successfully only by the intravenous or intrahepatic route. To assess the route of HEV transmission, 27 pigs were separated into nine groups of three pigs. Positive-control pigs were inoculated intravenously with swine HEV and served as the source of HEV for the other groups. Uninoculated contact pigs were placed in the positive-control group. On three consecutive days, naive pigs were inoculated using samples collected from the positive-control pigs at 9, 10, and 11 days postinoculation. The tonsils and nasal mucosa of each positive-control pig were swabbed and that swab was used to rub the tonsils and nasal and ocular mucosa of naive pigs. The positive-control pigs were also injected with bacterin, and the same needle was used to immediately inject naive pigs. Feces were collected from positive controls and fed by oral gavage to naive pigs. Weekly fecal and serum samples from each pig were tested for anti-HEV antibodies and HEV RNA. All positive control pigs shed the virus in feces; two pigs were viremic and seroconverted to anti-HEV. All contact control pigs shed the virus in feces; two seroconverted and one became viremic. One of three pigs in the fecal-oral exposure group shed the virus in feces and seroconverted. Pigs exposed to the contaminated needles or the tonsil and nasal secretion swabs remained negative. This is the first report of experimental fecal-oral transmission of HEV in swine. PMID- 15528695 TI - Mycobacterium africanum genotyping using novel spacer oligonucleotides in the direct repeat locus. AB - This study involves a first evaluation of 25 novel spacer oligonucleotides in addition to the 43 routine spacers for molecular characterization of a panel of 65 isolates of tubercle bacilli from different geographic origins that were initially classified as Mycobacterium africanum based on phenotypic characters. The 68-spacer format defined four additional patterns, and three groups were identified. The relatively homogeneous groups A1 and A2 included strains from West Africa, and A3-1 included strains from East Africa. The presence of deletion region RD9 confirmed the reclassification of the M. africanum subtype II spoligopattern within group A3-1 as Mycobacterium tuberculosis. These isolates may represent a diverging branch of M. tuberculosis in Africa. The use of new spacers also suggested an undergoing evolution of M. africanum subtype I in West Africa. Our results showed that the strain differentiation within the M. tuberculosis complex is improved by using novel spacers, and extensive studies using new-generation spoligotyping may be helpful to better understand the evolution of M. africanum. PMID- 15528696 TI - Characterization of ancestral Mycobacterium tuberculosis by multiple genetic markers and proposal of genotyping strategy. AB - Sixty-eight ancestral Mycobacterium tuberculosis isolates were previously identified by using the tuberculosis-specific deletion 1 (TbD1) PCR and mycobacterial interspersed-repetitive-unit-variable-number tandem repeat (MIRU VNTR) typing (Y. J. Sun, R. Bellamy, A. S. G. Lee, S. T. Ng, S. Ravindran, S.-Y. Wong, C. Locht, P. Supply, and N. I. Paton, J. Clin. Microbiol. 42:1986-1993, 2004). These TbD1(+) ancestral isolates were further characterized and typed in this study by IS6110 restriction fragment length polymorphism (RFLP) typing, VNTR typing using exact tandem repeats (VNTR-ETR), and spoligotyping of the direct repeat region. To our knowledge, this is the first characterization of this genogroup by multiple genetic markers based on a fairly large sample size. In this genogroup, all spoligotypes were characterized by the absence of spacers 29 to 32 and 34. In addition, VNTR-ETR typing could add further resolution to the clustered isolates identified by MIRU-VNTR, and the combination of MIRU-VNTR and VNTR-ETR, called MIRU-ETR, showed the highest discriminatory power for these strains compared to IS6110 RFLP typing and spoligotyping alone. However, MIRU-ETR appeared to still cluster some probably epidemiologically unrelated strains, as judged by IS6110 RFLP divergence. Therefore, a typing strategy based on stepwise combination of MIRU-ETR and IS6110 RFLP is proposed to achieve maximal discrimination for unrelated TbD1(+) strains. This typing strategy may be useful in areas where TbD1(+) ancestral strains are prevalent. PMID- 15528697 TI - Multilocus microsatellite typing system for Penicillium marneffei reveals spatially structured populations. AB - For eukaryotic pathogens that have low levels of genetic variation, multilocus microsatellite typing (MLMT) offers an accurate and reproducible method of characterizing genetic diversity. Here, we describe the application of an MLMT system to the emerging pathogenic fungus Penicillium marneffei. Isolates used for this study were those held in the culture collections of the Centraalbureau voor Schimmelcultures, Utrecht, The Netherlands, and the Chiang Mai University Department of Microbiology, Chang Mai, Thailand. High genetic diversity and extensive spatial structure were observed among clinical isolates, with the geographical area of origin for each isolate strongly correlating with the occurrence of two deeply divided clades. Within each clade, multilocus linkage associations were highly significant and could be explained by genetically differentiated populations or by an exclusively clonal reproductive mode, or both. Our results show that southeast Asian penicilliosis is caused by a fungus with a complex population genetic structure. Furthermore, this MLMT system generates digital data that can be easily queried against a centrally held database via the internet (http://pmarneffei.multilocus.net/); this provides a powerful epidemiological tool for analyzing the underlying parameters that are responsible for the emergence of P. marneffei in human immunodeficiency virus positive populations. PMID- 15528698 TI - Role of Cannomys badius as a natural animal host of Penicillium marneffei in India. AB - Infection by Penicillium marneffei in human immunodeficiency virus-positive patients in India has recently been described; the aim of our study was to survey wild rodents and their associated environment in order to identify the natural populations of this fungus. Surveys recovered P. marneffei from the internal organs of 10 (9.1%) of 110 bamboo rats (Cannomys badius) examined from Manipur state, India, an area endemic for penicilliosis marneffei. Identification of the isolates was based on a detailed study of their morphological characteristics, in vitro conversion to fission yeast form, and exoantigen tests. Multilocus microsatellite typing (MLMT) of the isolates revealed five genotypes. No genotypes were shared between sample sites, and all bamboo rats were infected with a single genotype within sample sites, demonstrating spatial genetic heterogeneity. One MLMT genotype was identical to that seen in a human isolate, suggesting that either coinfection from a common source or host-to-host transmission had occurred. This demonstrates the utility of an MLMT-based approach to elucidating the epidemiology of P. marneffei. PMID- 15528700 TI - Development and evaluation of an enzyme-linked immunosorbent assay for quantifying antibodies to Japanese encephalitis virus nonstructural 1 protein to detect subclinical infections in vaccinated horses. AB - Antibodies to Japanese encephalitis virus (JEV) nonstructural 1 (NS1) protein constitute a marker of natural JEV infection among populations vaccinated with inactivated JE vaccine. In Japan, with few recent human JE cases, the natural infection rate is critical to evaluate the necessity of continuing JE vaccination. A sensitive immunochemical staining method for detecting NS1 antibodies in individuals naturally and subclinically infected with JEV was previously established. Here, an enzyme-linked immunosorbent assay (ELISA) to detect NS1 antibodies in equine sera was developed and evaluated as an alternative to immunostaining. By this method, NS1 antigens contained in culture fluids from cells stably transfected with the NS1 and NS2A genes were captured by a rabbit anti-NS1 polyclonal antibody. Three nanograms per well of NS1 antigen, corresponding to 1:2 to 1:8 dilutions of the culture fluid, was sufficient for testing. ELISA values were obtained by a single-serum dilution (1:100), which correlated with ELISA titers obtained by an endpoint method. Under a tentative cutoff value (0.122) statistically calculated from NS1 antibody levels of horses in an area where JEV is not endemic, a high level of qualitative agreement (85.3%) was obtained between the ELISA and immunostaining methods. A significant correlation coefficient (0.799; P < 0.001) was also obtained between the two methods. Three experimentally infected horses seroconverted no later than 13 to 23 days postinfection, whereas 4 field horses infected during an epizootic remained positive for NS1 antibodies for at least 40 weeks. Our results indicate that the ELISA used here was sufficiently sensitive to detect subclinical infections in vaccinated equine populations. PMID- 15528699 TI - Borrelia species in host-seeking ticks and small mammals in northern Florida. AB - The aim of this study was to improve understanding of several factors related to the ecology and environmental risk of Borrelia infection in northern Florida. Small mammals and host-seeking adult ticks were collected at several sites, and specimens were tested for the presence of Borrelia species, primarily by PCR amplification. Tissues from some vertebrates and ticks were initially cultured in BSK-H medium to isolate spirochetes, but none were recovered. However, comparison of partial flagellin (flaB), 66-kDa protein (p66), and outer surface protein A (ospA) gene sequences from DNAs amplified from small mammals and ticks confirmed the presence of several Borrelia species. Borrelia lonestari DNA was detected among lone star ticks (Amblyomma americanum) at four sites. Borrelia burgdorferi sensu stricto strains were detected in all small mammal species tested and in A. americanum, Ixodes affinis, and Ixodes scapularis ticks. Borrelia bissettii was found in a cotton mouse and cotton rats and in I. affinis ticks. The study findings extend the known geographic distributions of B. lonestari in A. americanum and of B. burgdorferi sensu lato in A. americanum, I. affinis, I. scapularis, and small mammals to new sites in Florida. The presence of B. burgdorferi sensu stricto strains in host-seeking lone star ticks at two sites in Florida suggests that A. americanum should still be considered a possible vector of B. burgdorferi sensu lato. PMID- 15528701 TI - Outbreak of carbapenem-resistant Pseudomonas aeruginosa producing VIM-8, a novel metallo-beta-lactamase, in a tertiary care center in Cali, Colombia. AB - The prevalence of imipenem resistance among Pseudomonas aeruginosa isolates at a 195-bed tertiary care medical center in Cali, Colombia, rose from 2% in 1996 to 28% in 1997 and to over 40% in 2003. Many isolates showed high-level multiresistance, and phenotypic characterization suggested the spread of a predominant strain with minor variants. Sixty-six resistant isolates collected between February 1999 and July 2003 from hospitalized patients (n = 54) and environmental samples (n = 12) were subjected to a fuller analysis. Genetic fingerprints were compared by pulsed-field gel electrophoresis (PFGE) of SpeI digested genomic DNA, and bla(IMP) and bla(VIM) genes were sought by PCR. PFGE and serotyping indicated that 52 of the 66 isolates belonged to a single strain, with 82% similarity; the PFGE pattern for this organism was designated pattern A. Two further pairs of isolates represented single strains; the remaining nine isolates were unique, and in the case of one isolate, no satisfactory PFGE profile could be obtained. The pattern A isolates were mostly of serotype O12 and were highly resistant to imipenem (MICs, 32 to >256 microg/ml), with this resistance decreased eightfold or more in the presence of EDTA. They yielded amplicons with bla(VIM)-specific primers, and sequencing of DNA from a representative isolate revealed bla(VIM-8), a novel allele with three polymorphisms compared with the sequence of bla(VIM-2). Two of these nucleotide changes were silent, but the third determined a Thr139Ala substitution. Only 4 of 13 resistant isolates (2 clinical isolates and 2 environmental isolates) assigned to other PFGE types carried bla(VIM) alleles, whereas the others were less multiresistant and mostly had lower levels of imipenem resistance (MICs, < or =32 microg/ml) which was not significantly reduced by EDTA. No bla(IMP) alleles were detected. During 2003, when the environmental study was undertaken, serotype O12 isolates with bla(VIM) were recovered from sinks and stethoscopes in the most affected units, although not from the hands of staff; the problem declined once these reservoirs were disinfected and hygienic precautions were reinforced. PMID- 15528702 TI - Antimicrobial susceptibility testing of Acinetobacter spp. by NCCLS broth microdilution and disk diffusion methods. AB - Although both broth microdilution (BMD) and disk diffusion (DD) are listed by NCCLS as acceptable methods for testing Acinetobacter spp. for antimicrobial susceptibility, few studies have compared the results generated by the two methods. We tested 196 isolates of Acinetobacter spp. from nine U.S. hospitals and from the Centers for Disease Control culture collection by using BMD and DD and clinically appropriate antimicrobial agents. Categorical results for amikacin, ciprofloxacin, gatifloxacin, gentamicin, imipenem, levofloxacin, meropenem, tobramycin, and trimethoprim-sulfamethoxazole were comparable for the two methods: there was only one very major (VM) error, with tobramycin, and only one major (M) error, with meropenem, when DD results were compared with BMD results. However, VM errors were frequent with the beta-lactams and beta-lactam beta-lactam inhibitor combinations, while M errors were often observed with tetracyclines. For BMD, tests frequently exhibited subtle growth patterns that were difficult to interpret, especially for beta-lactams. If subtle growth (i.e., granular, small button, or "starry" growth) was considered positive, error rates between BMD and DD were unacceptably high for ampicillin-sulbactam (VM error, 9.8%; minor [m] error, 16.1%), piperacillin (VM error, 5.7%; m error, 13.5%), piperacillin-tazobactam (VM error, 9.3%; m error, 12.9%), ceftazidime (VM error, 6.2%; m error, 11.4%), cefepime (VM error, 6.2%; m error, 13.0%), cefotaxime (m error, 21.2%), ceftriaxone (m error, 23.3%), tetracycline (M error, 11.4%; m error, 32.1%), and doxycycline (M error, 2.6%). When subtle growth patterns were ignored, the agreement still did not achieve acceptable levels. To determine if the problems with BMD testing occurred in other laboratories, we sent frozen BMD panels containing beta-lactam drugs and nine isolates to six labs with experience in performing BMD and DD. Among these laboratories, cefepime MICs ranged from < or =8 to > or =32 microg/ml for four of the nine strains, confirming the problem in interpreting BMD results. Discrepancies between the categorical interpretations of BMD and DD tests were noted primarily with cefepime and piperacillin, for which the BMD results were typically more resistant. Clinical laboratories should be aware of these discrepancies. At present, there are no data to indicate which method provides more clinically relevant information. PMID- 15528704 TI - Diagnostic relevance of immunoglobulin G avidity for hepatitis A virus. AB - Diagnosis of acute hepatitis A virus (HAV) infection is based on the detection of HAV immunoglobulin M (IgM). However, IgM could be detected due to nonspecific polyclonal activation of the immune system. An avidity test for anti-HAV IgG was developed to distinguish acute infection, where low-avidity antibodies are detected, from immune reactivation. The assay was tested on 104 samples, including 11 sera from patients with past infection, 15 sera from patients with acute infection and 4 collected after recovery, 10 sera from vaccinated subjects, 4 sera from patients with suspected immune reactivation, and 60 unselected HAV IgM positive sera, collected over 1 year in a routine laboratory. The avidity index (AI) was expressed as percentage. The results were provided as the mean +/- one standard deviation. Patients with a history of prior infection had AIs of >70% (mean, 86% +/- 10), whereas the mean AI was 36% +/- 16 during acute HAV infection (P < 0.001). Within the first month after the onset of hepatitis, avidity was either noncalculable due to a very low IgG titer or <50%. In patients with immune reactivation, avidity was >70% (88% +/- 10%), a finding consistent with a prior infection. Among the 60 unselected sera, 35 (58%) had a noncalculable or <50% avidity, and most of them had a detectable HAV RNA, confirming HAV infection. In contrast, 16 (27%) had an avidity of >70%, and none was reverse transcription-PCR positive, suggesting immune reactivation. These 16 patients were significantly older than the others (50 +/- 16 years versus 26 +/- 14 years). The new anti-HAV IgG avidity assay we developed could improve HAV infection diagnosis, particularly in elderly patients. PMID- 15528703 TI - Genetic diversity of human pathogenic members of the Fusarium oxysporum complex inferred from multilocus DNA sequence data and amplified fragment length polymorphism analyses: evidence for the recent dispersion of a geographically widespread clonal lineage and nosocomial origin. AB - Fusarium oxysporum is a phylogenetically diverse monophyletic complex of filamentous ascomycetous fungi that are responsible for localized and disseminated life-threatening opportunistic infections in immunocompetent and severely neutropenic patients, respectively. Although members of this complex were isolated from patients during a pseudoepidemic in San Antonio, Tex., and from patients and the water system in a Houston, Tex., hospital during the 1990s, little is known about their genetic relatedness and population structure. This study was conducted to investigate the global genetic diversity and population biology of a comprehensive set of clinically important members of the F. oxysporum complex, focusing on the 33 isolates from patients at the San Antonio hospital and on strains isolated in the United States from the water systems of geographically distant hospitals in Texas, Maryland, and Washington, which were suspected as reservoirs of nosocomial fusariosis. In all, 18 environmental isolates and 88 isolates from patients spanning four continents were genotyped. The major finding of this study, based on concordant results from phylogenetic analyses of multilocus DNA sequence data and amplified fragment length polymorphisms, is that a recently dispersed, geographically widespread clonal lineage is responsible for over 70% of all clinical isolates investigated, including all of those associated with the pseudoepidemic in San Antonio. Moreover, strains of the clonal lineage recovered from patients were conclusively shown to genetically match those isolated from the hospital water systems of three U.S. hospitals, providing support for the hypothesis that hospitals may serve as a reservoir for nosocomial fusarial infections. PMID- 15528706 TI - Evaluation of a candidate international standard preparation for human anti Toxoplasma immunoglobulin G. AB - A freeze-dried human serum preparation containing immunoglobulin G (IgG) to Toxoplasma gondii was assessed for its suitability as an international reference reagent in an international collaborative study by 24 laboratories from 17 countries. This candidate standard was compared with the third international standard (IS) for human anti-Toxoplasma serum, TOXM, with the previous second IS, TOXS, and with a range of other serum samples. Samples were tested with the Sabin Feldman dye test and a range of agglutination assays and enzyme immunoassays. This study emphasizes the need for appropriate standards if intermethod agreement of estimates is to be achieved. On the basis of the results of this study, the preparation was established by the World Health Organization as the first IS for human anti-Toxoplasma IgG, with an assigned potency of 20 IU per ampoule of total anti-Toxoplasma antibodies. PMID- 15528705 TI - Detection of Campylobacter spp. in chicken fecal samples by real-time PCR. AB - A real-time PCR assay for detecting thermophilic Campylobacter spp. directly in chicken feces has been developed. DNA was isolated from fecal material by using magnetic beads followed by PCR with a prealiquoted PCR mixture, which had been stored at -18 degrees C. Campylobacter could be detected in less than 4 h, with a detection limit of 100 to 150 CFU/ml, in a fecal suspension. A bacterial internal control was added before DNA extraction to control both DNA isolation and the presence of PCR inhibitors in the samples. The assay was performed on 111 swab samples from a Danish surveillance program and compared to conventional culturing using selective enrichment. There was no statistically significant difference in performance between real-time PCR and culture by selective enrichment, and the diagnostic specificity was 0.96 with an agreement of 0.92. Therefore, the assay should be useful for screening poultry flocks for the presence of Campylobacter. PMID- 15528707 TI - Nocardia kruczakiae sp. nov., a pathogen in immunocompromised patients and a member of the "N. nova complex". AB - Molecular methodologies have become useful techniques for the identification of pathogenic Nocardia species and for the recognition of novel species that are capable of causing human disease. Two isolates recovered from immunocompromised patients were characterized as Nocardia nova by biochemical and susceptibility testing results. The restriction fragment length polymorphism (RFLP) patterns obtained by restriction endonuclease analysis (REA) of an amplified portion of the heat shock protein gene were identical to those obtained with the type strain of N. nova. REA of an amplified portion of the 16S rRNA gene showed RFLP patterns that were unlike those obtained for the type strain of N. nova but that were similar to those obtained for the type strains of N. africana and N. veterana. Subsequent sequencing of a portion of the 16S rRNA gene produced identical results for the two patient isolates. Sequence analysis of 1,352-bp portions of the 16S rRNA gene indicated that these isolates were 99.8% similar to the recently described species N. veterana but were only 99.3, 98.1, and 98.1% similar to the type strains of N. africana, N. nova, and N. vaccinii, respectively. DNA-DNA hybridization studies confirmed that the two patient isolates belonged to the same species but were not closely related to N. africana, N. nova, N. vaccinii, or N. veterana. The patient isolates have been designated N. kruczakiae sp. nov. Because N. africana, N. veterana, and the new species are not readily differentiated from N. nova by phenotypic methods alone, the designation "N. nova complex" can be used to designate isolates such as these that phenotypically resemble N. nova but that have not been definitively characterized by 16S rRNA gene sequencing or DNA-DNA hybridization. PMID- 15528708 TI - Distribution of serogroups and genotypes among disease-associated and carried isolates of Neisseria meningitidis from the Czech Republic, Greece, and Norway. AB - The distribution of serogroups and multilocus sequence types (STs) in collections of disease-associated and carried meningococci from the period 1991 to 2000 in three European countries (the Czech Republic, Greece, and Norway) was investigated. A total of 314 patient isolates and 353 isolates from asymptomatic carriers were characterized. The frequency distributions of serogroups and clone complexes differed among countries and between disease and carrier isolate collections. Highly significant differentiation was seen at each housekeeping locus. A marked positive association of serogroup C with disease was evidenced. The ST-11 complex was strongly positively associated with disease; associations for other clone complexes were weaker. The genetic diversity of the clone complexes differed. A single ST dominated the ST-11 clone complex, while the ST 41/44 complex exhibited greater levels of diversity. These data robustly demonstrated differences in the distribution of meningococcal genotypes in disease and carrier isolates and among countries. Further, they indicated that differences in genotype diversity and pathogenicity exist between meningococcal clone complexes. PMID- 15528709 TI - Emergence, spread, and characterization of phage variants of epidemic methicillin resistant Staphylococcus aureus 16 in England and Wales. AB - Epidemic methicillin-resistant Staphylococcus aureus 16 (EMRSA-16) and EMRSA-15 are the two most important and prevalent EMRSA strains found in the United Kingdom and have also been found in a number of European countries and the United States. We describe for the first time the spread of an EMRSA strain (EMRSA-16) from its point of origin in one hospital to the surrounding hospitals and regions over the following 2 years. In the first 18 months after its original appearance, 136 hospitals referred EMRSA-16 isolates for typing, and interhospital and intraregional spread were reported: it was more prevalent in males between 60 and 80 years old and was isolated from sputum and throat more often than EMRSA-15. Important characteristics, e.g., carriage of the enterotoxin A (sea) and toxic shock syndrome toxin (tst) genes and production of urease, are described. Phage variant strains of EMRSA-16 which share some of the characteristics of the classical strain, including toxin carriage and urease production, emerged, but without genotypic investigations, their relationship could only be inferred. A total of 129 clinical isolates from 52 hospitals, collected between March 1998 and April 1999 and representing classical EMRSA-16 (49 isolates) or phage variants (80 isolates), were compared by phage typing, pulsed-field gel electrophoresis (PFGE) following SmaI macrorestriction, antimicrobial susceptibility testing, urease production, and PCR detection of toxin gene carriage. PFGE analysis revealed 29 profiles, A1 to A29, with A1 representing the prototypic strain, NCTC 13143. All other profiles differed from A1 by 1 to 6 bands, but some differed from each other by up to 10 bands. PMID- 15528710 TI - New multiple antigenic peptide-based enzyme immunoassay for detection of simian immunodeficiency virus infection in nonhuman primates and humans. AB - Infections with human immunodeficiency virus types 1 and 2 (HIV-1 and HIV-2, respectively) are zoonotic infections. In Africa, the potential exists for additional cross-species transmissions from at least 33 different species of simian immunodeficiency virus (SIV)-infected nonhuman primates (NHPs) through hunting and butchering of these animals for food. Here we describe a highly sensitive and specific enzyme immunoassay (EIA) with chemically modified, multiple antigenic peptides (MAPs) developed for the detection and discrimination of antibodies to SIV genetic lineages. The SIV EIA was developed by using a comprehensive array of MAPs covering two envelope gene regions from all of the SIV lineages for which env sequences were available. Assay sensitivity was evaluated by using 63 plasma or serum samples obtained from primates naturally or experimentally infected with SIVs from 10 genetic lineages. Assay specificity was determined by using 97 known SIV-negative plasma specimens from these same species. Also used in the evaluations were 369 human samples: 198 HIV seronegative, 170 HIV-1 and/or HIV-2 seropositive, and 1 from a human SIVsm infection. Overall assay sensitivity and specificity were 100% with both immunodominant region (IDR) and V3 region MAPs. Although SIV env sequences from talapoin monkeys were not available for specific MAP inclusion, 5 (100%) of 5 SIVtal-infected samples were detected through cross-reactivity with other SIV IDR MAPs used in the assay. The one human SIVsm infection was identified. In conclusion, our SIV MAP EIA proved to be highly sensitive and specific for detecting SIV infections in NHPs and humans. As shown with SIV-infected talapoin monkeys, this assay has the potential to detect previously unidentified SIV strains and should be suitable for sentinel surveillance for potential new cross species transmissions of SIVs to humans. PMID- 15528711 TI - Analysis of vaginal acetic acid in patients undergoing treatment for bacterial vaginosis. AB - A "gold standard" method for the diagnosis of bacterial vaginosis (BV) is lacking. The clinical criteria described by the Amsel technique are subjective and difficult to quantify. Alternatively, the reading of Gram-stained vaginal smears by scoring techniques such as those that use the Nugent or Hay-Ison scoring systems is again subjective, requires expert personnel to perform the reading, and is infrequently used clinically. Recently, a new diagnostic device, the Osmetech Microbial Analyzer--Bacterial Vaginosis (OMA-BV), which determines a patient's BV status on the basis of measurement of the amount of acetic acid present in a vaginal swab specimen, was approved by the Food and Drug Administration. The present study uses the conducting polymer gas-sensing technology of OMA-BV to measure the concentration of acetic acid in the headspace above vaginal swab specimens from patients undergoing treatment for BV with metronidazole. In 97.8% of the cases the level of acetic acid detected fell sharply during the treatment period, crossing from above to below the diagnostic threshold of 900 ppm. The diagnosis obtained on the basis of the level of vaginal acetic acid was compared with the diagnoses obtained by use of the Amsel criteria and the Nugent scoring system both at the time of initial entry into the study and at the repeat samplings on days 7 and 14. The results obtained with OMA-BV showed overall agreements compared with the results of the Amsel and Nugent tests of 98 and 94%, respectively, for the 34 patients monitored through the treatment process. This provides further evidence that the measurement of vaginal acetic acid by headspace analysis with conducting polymer sensors is a valid alternative to present tests for the diagnosis of BV. PMID- 15528712 TI - characterization of bacterial community diversity in cystic fibrosis lung infections by use of 16s ribosomal DNA terminal restriction fragment length polymorphism profiling. AB - Progressive loss of lung function resulting from the inflammatory response to bacterial colonization is the leading cause of mortality in cystic fibrosis (CF) patients. A greater understanding of these bacterial infections is needed to improve lung disease management. As culture-based diagnoses are associated with fundamental drawbacks, we used terminal restriction fragment (T-RF) length polymorphism profiling and 16S rRNA clone data to characterize, without prior cultivation, the bacterial community in 71 sputa from 34 adult CF patients. Nineteen species from 15 genera were identified in 53 16S rRNA clones from three patients. Of these, 15 species have not previously been reported in CF lung infections and many were species requiring strict anaerobic conditions for growth. The species richness and evenness were determined from the T-RF length and volume for the 71 profiles. Species richness was on average 13.3 +/- 7.9 per sample and 13.4 +/- 6.7 per patient. On average, the T-RF bands of the lowest and highest volumes represented 0.6 and 59.2% of the total volume in each profile, respectively. The second through fifth most dominant T-RF bands represented 15.3, 7.5, 4.7, and 2.8% of the total profile volume, respectively. On average, the remaining T-RF bands represented 10.2% of the total profile volume. The T-RF band corresponding to Pseudomonas aeruginosa had the highest volume in 61.1% of the samples. However, 18 other T-RF band lengths were dominant in at least one sample. In conclusion, this reveals the enormous complexity of bacteria within the CF lung. Although their significance is yet to be determined, these findings alter our perception of CF lung infections. PMID- 15528713 TI - Effects of intrapartum penicillin prophylaxis on intestinal bacterial colonization in infants. AB - Early-onset group B streptococcal (GBS) infections remain a leading cause of morbidity and mortality in infants. To prevent the vertical transmission of GBS and neonatal GBS infection, guidelines recommend intrapartum penicillin or amoxicillin prophylaxis. This intrapartum antibiotic prophylaxis (IAP) is suspected to favor colonization by antibiotic-resistant bacteria. However, the effects of this prophylaxis on the patterns of acquisition of gastrointestinal bacterial flora in infants have never been studied. We collected stool samples from 3-day-old infants born to mothers who received intrapartum amoxicillin (antibiotic-exposed group; n = 25) and to untreated mothers (non-antibiotic exposed group; n = 25). The groups were matched for factors known to affect intestinal microbial colonization: gestational age, type of delivery, and type of feeding. Qualitative and quantitative differential analyses of the bacterial flora in stool samples were performed. Similar numbers of infants in the non antibiotic-exposed and antibiotic-exposed groups were colonized by aerobic bacteria and amoxicillin-resistant enterobacteria (75 and 77%, respectively) (P = 0.79). In contrast, significantly fewer infants in the antibiotic-exposed group than in the non-antibiotic-exposed group were colonized by anaerobic bacteria, especially Clostridium (12 and 40%, respectively) (P < 0.05). Regarding intestinal bacterial colonization, the differences between antibiotic-exposed and non-antibiotic-exposed infants were remarkably few. The only statistically significant effect was the reduced initial bacterial colonization by Clostridium in the antibiotic-exposed group. In our study, the use of IAP did not favor colonization by beta-lactam-resistant bacteria. However, further evaluations are required to highlight the potential risks of the widespread use of antibiotics to prevent early-onset GBS infection. PMID- 15528714 TI - Real-time quantitative PCR assays for detection and monitoring of pathogenic human viruses in immunosuppressed pediatric patients. AB - A panel of 23 real-time PCR assays based on TaqMan technology has been developed for the detection and monitoring of 16 different viruses and virus families including human polyomaviruses BK virus and JC virus, human herpesviruses 6, 7, and 8, human adenoviruses, herpes simplex viruses 1 and 2, varicella-zoster virus, cytomegalovirus, Epstein-Barr virus, parvovirus B19, influenza A and B viruses, parainfluenza viruses 1 to 3, enteroviruses, and respiratory syncytial virus. The test systems presented have a broad dynamic range and display high sensitivity, reproducibility, and specificity. Moreover, the assays allow precise quantification of viral load in a variety of clinical specimens. The ability to use uniform PCR conditions for all assays permits simultaneous processing and detection of many different viruses, thus economizing the diagnostic work. Our observations based on more than 50,000 assays reveal the potential of the real time PCR tests to facilitate early diagnosis of infection and to monitor the kinetics of viral proliferation and the response to treatment. We demonstrate that, in immunosuppressed patients with invasive virus infections, surveillance by the assays described may permit detection of increasing viral load several days to weeks prior to the onset of clinical symptoms. In virus infections for which specific treatment is available, the quantitative PCR assays presented provide reliable diagnostic tools for timely initiation of appropriate therapy and for rapid assessment of the efficacy of antiviral treatment strategies. PMID- 15528715 TI - Assessment of the target-capture PCR hepatitis B virus (HBV) DNA quantitative assay and comparison with commercial HBV DNA quantitative assays. AB - Recent clinical studies suggest that hepatitis B virus (HBV) load and genotype may be independent predictors of responses to antiviral therapies. However, it is difficult for clinicians to accurately determine viral loads in patient samples because results--both the values and the units of measure--can vary greatly among different tests. Accordingly, the World Health Organization (WHO) has produced the first international standard for HBV DNA for nucleic acid amplification technology (NAT) assays. In the present study, we describe the performance of the target-capture PCR HBV DNA quantitative assay for the quantitation of HBV DNA in clinical samples and reference panels. The range of quantitation was between 50 and 10(10) IU/ml. The sensitivity and accuracy of the target-capture PCR assay were demonstrated by using the HBV panel from Quality Control for Medical Diagnostics (QCMD) and the WHO HBV DNA standard. The target-capture PCR assay quantitated the six genotype A members of the QCMD panel and dilutions of the WHO HBV DNA standard within an accuracy of 74 to 142%. Compared to current serological methods, the assay offers window period reductions of 19 days prior to HBV surface antigen and 26 days prior to HBV e antigen detection. The target capture PCR assay was also compared with four commercially available NAT assays, and the various units of measure were standardized with respect to the international units of the WHO HBV DNA standard. The target-capture PCR assay is a sensitive, accurate, high-throughput, rapid, and reproducible assay for the determination of HBV loads. PMID- 15528716 TI - Development of a PCR-restriction fragment length polymorphism assay for the epidemiological analysis of Shiga toxin-producing Escherichia coli. AB - Six characteristic regions (I to VI) were identified in Shiga toxin 2 (Stx2) phages (T. Sato, T. Shimizu, M. Watarai, M. Kobayashi, S. Kano, T. Hamabata, Y. Takeda, and S. Yamasaki, Gene 309:35-48, 2003). Region V, which is ca. 10 kb in size and is located in the upstream region of the Stx operons, includes the most distinctive region among six Stx phages whose genome sequences have been determined. In this study, we developed a PCR-restriction fragment length polymorphism (RFLP) assay for the epidemiological analysis of Shiga toxin producing Escherichia coli (STEC) on the basis of the diversity of region V. When region V was amplified by long and accurate-PCR (LA-PCR) with five control E. coli strains carrying six different Stx phages such as E. coli strains C600 (Stx1 phage), C600 (933W phage), C600 (Stx2 phage-I), C600 (Stx2 phage-II), and O157:H7 Sakai strain RIMD0509952 (VT1-Sakai phage and VT2-Sakai phage), an expected size of the band was obtained. Restriction digest of each PCR product with BglI or EcoRV also gave the expected sizes of banding patterns and discriminated the RFLPs of five control strains. When a total of 204 STEC O157 strains were analyzed by LA-PCR, one to three bands whose sizes ranged from 8.2 to 14 kb were obtained. Two STEC O157 strains, however, did not produce any bands. Subsequent restriction digest of the PCR products with BglI or EcoRV differentiated the RFLPs of 202 STEC O157 strains into 24 groups. The RFLP patterns of pulsed-field gel electrophoresis (PFGE) of representative strains of STEC O157 divided into 24 groups were well correlated with those of PCR-RFLP when STEC O157 strains were isolated in the same time period and in the close geographic area. To evaluate the PCR-RFLP assay developed here, ten strains, each isolated from four different outbreaks in different areas in Japan (Tochigi, Hyogo, Aichi, and Fukuoka prefecture), were examined to determine whether the strains in each group showed the same RFLP patterns in the PCR-RFLP assay. In accordance with the results of PFGE except for strains isolated in an area (Fukuoka), which did not produce any amplicon, ten strains in each group demonstrated the same RFLP pattern. Taken together, these data suggest that the PCR-RFLP based on region V is as useful as PFGE but perhaps more simple and rapid than PFGE for the molecular epidemiological analysis of STEC strains during sporadic and common source outbreaks. PMID- 15528717 TI - Genetic recombination between two genotypes of genogroup III bovine noroviruses (BoNVs) and capsid sequence diversity among BoNVs and Nebraska-like bovine enteric caliciviruses. AB - To determine the genogroups and genotypes of bovine enteric caliciviruses (BECVs) circulating in calves, we determined the complete capsid gene sequences of 21 BECVs. The nucleotide and predicted amino acid sequences were compared phylogenetically with those of known human and animal enteric caliciviruses. Based on these analyses, 15 BECVs belonged to Norovirus genogroup III and genotype 2 (GIII/2) and were genetically distinct from human Norovirus GI and GII. Six BECVs had capsid gene sequences similar to that of the unclassified Nebraska (NB)-like BECV. The 15 bovine noroviruses (BoNVs) were more closely related to Bo/NLV/Newbury-2/76/UK (GIII/2) and other known genotype 2 BoNVs than to genotype 1 Bo/NLV/Jena/80/DE. The BoNV Bo/CV521-OH/02/US showed high nucleotide and amino acid identities (84 and 94%, respectively) with the capsid gene of Bo/NLV/Newbury-2/76/UK, whereas the nucleotide and amino acid sequences of the RNA polymerase gene were more closely related to those of Bo/NLV/Jena/80/DE (77 and 87% identities, respectively) than to those of Bo/NLV/Newbury-2/76/UK (69 and 69% identities, respectively), suggesting that Bo/CV521-OH/02/US is a genotype 1-2 recombinant. Gene conversion analysis by the recombinant identification program and SimPlot also predicted that Bo/CV521 OH/02/US was a recombinant. Six NB-like BECVs shared 88 to 92% nucleotide and 94 to 99.5% amino acid identities with the NB BECV in the capsid gene. The results of this study demonstrate genetic diversity in the capsid genes of BECVs circulating in Ohio veal calves, provide new data for coinfections with distinct BECV genotypes or genogroups, and describe the first natural BoNV genotype 1-2 recombinant, analogous to the previously reported human norovirus recombinants. PMID- 15528718 TI - Validation of the use of Middlebrook 7H10 agar, BACTEC MGIT 960, and BACTEC 460 12B media for testing the susceptibility of Mycobacterium tuberculosis to levofloxacin. AB - Levofloxacin, the active l-isomer of the quinolone ofloxacin, is now widely accepted for treatment of multidrug-resistant tuberculosis. Because the drug is now widely used, we sought to establish susceptibility test conditions for Mycobacterium tuberculosis against levofloxacin by the traditional reference method, agar proportion (AP), the commonly used BACTEC 460 radiometric system, and the newer BACTEC MGIT 960 method. To determine the stability of levofloxacin in the two newer test systems (BACTEC 460 and BACTEC MGIT 960), media containing subinhibitory levels of levofloxacin were prepared and stored at 4 and 37 degrees C for 14 days. The stored media were inoculated with H37Rv, and the drug activity was compared to freshly prepared media. Results show that levofloxacin is stable over the course of testing. Next, optimum levofloxacin test concentrations were determined for AP, BACTEC 460, and BACTEC MGIT 960 methods. MICs were determined for 32 pan-susceptible isolates of M. tuberculosis obtained from presumably untreated patients and 14 quinolone-resistant isolates. The levofloxacin resistant strains either were isolated from patients who remained culture positive despite treatment with a quinolone agent (six strains) or contained known mutations in gyrA (eight strains). Levofloxacin MICs resulted in a bimodal pattern with values for resistant strains consistently higher than those for pan susceptible strains. Results show that levofloxacin concentrations of 2 microg/ml (BACTEC 460 and BACTEC MGIT 960) and 1 microg/ml (AP) inhibited the growth of all pan-susceptible strains while permitting the growth of all levofloxacin-resistant strains. Confirmatory tests with a subset of pan-susceptible and levofloxacin resistant isolates validated the selected test concentrations. PMID- 15528719 TI - Type III secretion phenotypes of Pseudomonas aeruginosa strains change during infection of individuals with cystic fibrosis. AB - Pseudomonas aeruginosa is a frequent cause of respiratory exacerbations in individuals with cystic fibrosis. An important virulence determinant of this pathogen is its type III protein secretion system. In this study, the type III secretion properties of 435 P. aeruginosa respiratory isolates from 56 chronically infected individuals with cystic fibrosis were investigated. Although it had been previously reported that 75 to 90% of P. aeruginosa isolates from patients with hospital-acquired pneumonia secreted type III proteins, only 12% of isolates from cystic fibrosis patients did so, with nearly all of these isolates secreting ExoS and ExoT but not ExoU. Despite the low overall prevalence of type III protein-secreting isolates, at least one secreting isolate was cultured from one-third of cystic fibrosis patients. Interestingly, the fraction of cystic fibrosis patient isolates capable of secreting type III proteins decreased with duration of infection. Although 90% of isolates from the environment, the presumed reservoir for the majority of P. aeruginosa strains that infect patients with cystic fibrosis, secreted type III proteins, only 49% of isolates from newly infected children, 18% of isolates from chronically infected children, and 4% of isolates from chronically infected adults with cystic fibrosis secreted these proteins. Within individual patients, isolates of clonal origin differed in their secretion phenotypes, indicating that as strains persisted in cystic fibrosis patient airways, their type III protein secretion properties changed. Together, these findings indicate that following infection of cystic fibrosis patient airways, P. aeruginosa strains gradually change from a type III protein secretion positive phenotype to a secretion-negative phenotype. PMID- 15528720 TI - Carious dentine provides a habitat for a complex array of novel Prevotella-like bacteria. AB - Previous analysis of the microbiology of advanced caries by culture and real-time PCR emphasized the high incidence and abundance of gram-negative anaerobic species, particularly Prevotella-like bacteria. The diversity of Prevotella-like bacteria was further explored by analyzing pooled bacterial DNA from lesions of carious dentine. This was achieved by amplification of a region of the 16S ribosomal DNA with a Prevotella genus-specific forward primer and a universal bacterial reverse primer, followed by cloning and sequencing. Cultured Prevotella species commonly associated with oral tissues constituted only 12% of the Prevotella clones isolated from advanced carious lesions. The remaining 88% consisted of a diverse range of phylotypes. These included five clusters of previously recognized but uncultured oral Prevotella spp. and a major cluster containing Prevotella-like bacteria most closely related to uncharacterized rumen bacteria. Cluster-specific primers were designed, and the numbers of bacteria within clusters were quantified by real-time PCR, confirming the abundance of these organisms. The data indicated that advanced dental caries provides a unique environment for a complex array of novel and uncultured Prevotella and Prevotella like bacteria which, in some cases, may dominate the diverse polymicrobial community associated with the disease. PMID- 15528721 TI - Usefulness of routine epicardial pacing wire culture for early prediction of poststernotomy mediastinitis. AB - Poststernotomy mediastinitis (PSM) is one of the most serious complications of cardiac surgery, and its associated morbidity and mortality demand early recognition for emergency therapy. In this study, we investigated the usefulness of epicardial pacing wire (EPW) cultures for the prediction of PSM. Among 2,200 patients who underwent a cardiac surgical procedure at our hospital between 1 January 1999 and 31 December 2001, 82 (3.7%) had PSM; Staphylococcus aureus was the organism (45.1%) most frequently isolated at the time of surgical debridement. EPWs from 1,607 (73.0%) patients, 73 (4.5%) of whom developed PSM, were cultured. EPW cultures from 466 (29.0%) were positive, most often (74.9%) for coagulase-negative Staphylococci. EPW cultures were truly positive in 26 cases, truly negative in 1,106 cases, falsely positive in 428 cases, and falsely negative in 47 cases (with sterile cultures in 35 cases and a culture positive for an organism different from that isolated at the time of debridement in 12 cases). EPW culture had a positive predictive value of only 5.7% and a high negative predictive value (95.9%) for the diagnosis of PSM, with an accuracy of 70.4%. However, the likelihood ratio of positive (1.27) and negative (0.89) tests indicated only small changes in pretest-to-posttest probability. Therefore, a strategy of routine culture of EPWs to predict PSM seems questionable. PMID- 15528722 TI - Quantification of Leishmania infantum DNA by a real-time PCR assay with high sensitivity. AB - A real-time PCR was developed to quantify Leishmania infantum kinetoplast DNA and optimized to reach a sensitivity of 0.0125 parasites/ml of blood. In order to analyze the incidence of heterogeneity and number of minicircles, we performed comparative PCR by using the Leishmania DNA polymerase gene as a reporter. Assays performed in both promastigote and amastigote stages showed variations among different L. infantum and Leishmania donovani strains and the stability of the minicircle numbers for a particular strain. Analysis of blood samples from a patient who presented with Mediterranean visceral leishmaniasis confirmed the reliability of such an assay for Leishmania quantification in biological samples and allowed an estimation of positivity thresholds of classical tests used for direct diagnosis of the disease; positivity thresholds were in the range of 18 to 42, 0.7 to 42, and 0.12 to 22.5 parasites/ml for microscopic examination, culture, and conventional PCR, respectively. At the time of diagnosis, parasitemia could vary by a wide range (32 to 188,700 parasites/ml, with a median of 837 parasites/ml), while in bone marrow, parasite load was more than 100 parasites per 10(6) nucleated human cells. After successful therapy, parasitemia levels remain lower than 1 parasite/ml. In the immunocompromised host, relapses correlate with an increase in the level of parasitemia, sometimes scanty, justifying the need for assays with high sensitivity. Such sensitivity allows the detection of Leishmania DNA in the blood of 21% of patients with no history of leishmaniasis living in the Marseilles area, where leishmaniasis is endemic. This technique may be useful for epidemiologic and diagnostic purposes, especially for the quantification of parasitemia at low levels during posttherapy follow-up. PMID- 15528723 TI - Metallo-beta-lactamase-producing gram-negative bacilli: laboratory-based surveillance in cooperation with 13 clinical laboratories in the Kinki region of Japan. AB - A total of 19,753 strains of gram-negative rods collected during two 6-month periods (October 2000 to March 2001 and November 2001 to April 2002) from 13 clinical laboratories in the Kinki region of Japan were investigated for the production of metallo-beta-lactamases (MBLs). MBLs were detected in 96 (0.5%) of the 19,753 isolates by the broth microdilution method, the 2-mercaptopropionic acid inhibition test, and PCR and DNA sequencing analyses. MBL-positive isolates were detected in 9 of 13 laboratories, with the rate of detection ranging between 0 and 2.6% for each laboratory. Forty-four of 1,429 (3.1%) Serratia marcescens, 22 of 6,198 (0.4%) Pseudomonas aeruginosa, 21 of 1,108 (1.9%) Acinetobacter spp., 4 of 544 (0.7%) Citrobacter freundii, 3 of 127 (2.4%) Providencia rettgeri, 1 of 434 (0.2%) Morganella morganii, and 1 of 1,483 (0.1%) Enterobacter cloacae isolates were positive for MBLs. Of these 96 MBL-positive strains, 87 (90.6%), 7 (7.3%), and 2 (2.1%) isolates carried the genes for IMP-1-group MBLs, IMP-2-group MBLs, and VIM-2-group MBLs, respectively. The class 1 integrase gene, intI1, was detected in all MBL-positive strains, and the aac (6')-Ib gene was detected in 37 (38.5%) isolates. Strains with identical PCR fingerprint profiles in a random amplified polymorphic DNA pattern analysis were isolated successively from five separate hospitals, suggesting the nosocomial spread of the organism in each hospital. In conclusion, many species of MBL-positive gram-negative rods are distributed widely in different hospitals in the Kinki region of Japan. The present findings should be considered during the development of policies and strategies to prevent the emergence and further spread of MBL-producing bacteria. PMID- 15528724 TI - Occurrence of extended-spectrum beta-lactamases in members of the genus Shigella in the Republic of Korea. AB - A nationwide survey was carried out in Korea to assess the prevalence of Shigella strains producing extended-spectrum beta-lactamases (ESBLs). From 1991 to 2002, 5,911 clinical strains were isolated and screened for resistance to extended spectrum cephalosporins. Twenty of the Shigella isolates were ESBL positive, based on the synergistic effects between clavulanate and selected beta-lactams (ceftazidime and cefotaxime). Nucleotide sequence analysis of these isolates revealed that they harbored bla(TEM-19) (eight isolates), bla(TEM-15) (five isolates), bla(TEM-52) (six isolates), bla(TEM-17) (one isolate), bla(TEM-20) (one isolate), and bla(CTX-M-14) (three isolates). All the ESBL-encoding genes in this study were carried in conjugable plasmids. Thus, TEM-19, TEM-15, TEM-52, and CTX-M-14 beta-lactamases can be considered common Korean ESBL types in Shigella sonnei and are probably transmitted through interspecies spread between medical facilities and the community in Korea. This is the first report of the presence of TEM-17, TEM-19, and TEM-20 in Korea and in S. sonnei. PMID- 15528725 TI - Selective discrimination of Listeria monocytogenes epidemic strains by a mixed genome DNA microarray compared to discrimination by pulsed-field gel electrophoresis, ribotyping, and multilocus sequence typing. AB - Listeria monocytogenes can cause serious illness in humans, and subsequent epidemiological investigation requires molecular characterization to allow the identification of specific isolates. L. monocytogenes is usually characterized by serotyping and is subtyped by using pulsed-field gel electrophoresis (PFGE) or ribotyping. DNA microarrays provide an alternative means to resolve genetic differences among isolates, and unlike PFGE and ribotyping, microarrays can be used to identify specific genes associated with strains of interest. Twenty strains of L. monocytogenes representing six serovars were used to generate a shotgun library, and subsequently a 629-probe microarray was constructed by using features that included only potentially polymorphic gene probe sequences. Fifty two strains of L. monocytogenes were genotyped by using the condensed array, including strains associated with five major listeriosis epidemics. Cluster analysis of the microarray data grouped strains according to phylogenetic lineage and serotype. Most epidemiologically linked strains were grouped together, and subtyping resolution was the same as that with PFGE (using AscI and ApaI) and better than that with multilocus sequence typing (using six housekeeping genes) and ribotyping. Additionally, a majority of epidemic strains were grouped together within phylogenetic Division I. This epidemic cluster was clearly distinct from the two other Division I clusters, which encompassed primarily sporadic and environmental strains. Discriminant function analysis allowed identification of 22 probes from the mixed-genome array that distinguish serotypes and subtypes, including several potential markers that were distinct for the epidemic cluster. Many of the subtype-specific genes encode proteins that likely confer survival advantages in the environment and/or host. PMID- 15528726 TI - Dual-probe assay for rapid detection of drug-resistant Mycobacterium tuberculosis by real-time PCR. AB - Mutations in particular nucleotides of genes coding for drug targets or drug converting enzymes lead to drug resistance in Mycobacterium tuberculosis. For rapid detection of drug-resistant M. tuberculosis in clinical specimens, a simple and applicable method is needed. Eight TaqMan minor groove binder (MGB) probes, which discriminate one-base mismatches, were designed (dual-probe assay with four reaction tubes). The target of six MGB probes was the rpoB gene, which is involved in rifampin resistance; five probes were designed to detect for mutation sites within an 81-bp hot spot of the rpoB gene, and one probe was designed as a tuberculosis (TB) control outside the rpoB gene hot-spot. We also designed probes to examine codon 315 of katG and codon 306 of embB for mutations associated with resistance to isoniazid and ethambutol, respectively. Our system was M. tuberculosis complex specific, because neither nontuberculous mycobacteria nor bacteria other than mycobacteria reacted with the system. Detection limits in direct and preamplified analyses were 250 and 10 fg of genomic DNA, respectively. The system could detect mutations of the rpoB, katG, and embB genes in DNAs extracted from 45 laboratory strains and from sputum samples of 27 patients with pulmonary TB. This system was much faster (3 h from DNA preparation) than conventional drug susceptibility testing (3 weeks). Results from the dual-MGB probe assay were consistent with DNA sequencing. Because the dual-probe assay system is simple, rapid, and accurate, it can be applied to detect drug-resistant M. tuberculosis in clinical laboratories. PMID- 15528727 TI - Characterization of "Candidatus piscichlamydia salmonis" (order Chlamydiales), a chlamydia-like bacterium associated with epitheliocystis in farmed Atlantic salmon (Salmo salar). AB - To characterize intracellular gram-negative bacteria associated with epitheliocystis in farmed Atlantic salmon (Salmo salar), gills with proliferative lesions were collected for histopathology, conventional transmission and immunoelectron microscopy, in situ hybridization, and DNA extraction during epitheliocystis outbreaks in Ireland and Norway in 1999 and 2000, respectively, and compared by ultrastructure and immunoreactivity to nonproliferative gills from Ireland archived in 1995. Genomic DNA from proliferative gills was used to amplify 16S ribosomal DNA (rDNA) for molecular phylogenetic analyses. Epitheliocystis inclusions from proliferative gills possessed variably elongate reticulate bodies, examples of binary fission, and vacuolated and nonvacuolated intermediate bodies, whereas inclusions in nonproliferative gills had typical chlamydial developmental stages plus distinctive head-and-tail cells. Immunogold processing using anti-chlamydial lipopolysaccharide antibody labeled reticulate bodies from proliferative and nonproliferative gills. 16S rDNA amplified directly from Irish (1999) and Norwegian (2000) gill samples demonstrated 99% nucleotide identity, and riboprobes transcribed from cloned near-full-length 16S rDNA amplicons from Norwegian gills hybridized with inclusions in proliferative lesions from Irish (1999) and Norwegian (2000) sections. A 1,487-bp consensus 16S rRNA gene sequence representing the chlamydia-like bacterium (CLB) from proliferative gills had the highest percent nucleotide identity with endosymbionts of Acanthamoeba spp. (order Chlamydiales). Molecular phylogenetic relationships inferred from 16S rRNA gene sequences using distance and parsimony indicated that the CLB from proliferative gills branched with members of the order Chlamydiales. "Candidatus Piscichlamydia salmonis" is proposed for the CLB associated with epitheliocystis from proliferative gills of Atlantic salmon, which exhibits developmental stages different from those identified in nonproliferative gills. PMID- 15528728 TI - Porphyromonas uenonis sp. nov., a pathogen for humans distinct from P. asaccharolytica and P. endodontalis. AB - Three Porphyromonas species (Porphyromonas asaccharolytica, P. endodontalis, and the novel species that is the subject of the present report, P. uenonis) are very much alike in terms of biochemical characteristics, such as enzyme profiles and cellular fatty acid contents. P. asaccharolytica is distinguished from the other two species by virtue of production of alpha-fucosidase and glyoxylic acid positivity. The novel species is difficult to differentiate from P. endodontalis phenotypically and was designated a P. endodontalis-like organism for some time. However, P. endodontalis is recovered almost exclusively from oral sources and also grows poorly on Biolog Universal Agar, both characteristics that are in contrast to those of the other two organisms. Furthermore, P. uenonis is glycerol positive in the Biolog AN Microplate system. Both P. asaccharolytica and P. uenonis are positive by 13 other tests in the Biolog system, whereas P. endodontalis is negative by all of these tests. P. asaccharolytica grew well in both solid and liquid media without supplementation with 5% horse serum, whereas the other two species grew poorly without supplementation. Sequencing of 16S rRNA revealed about 10% divergence between the novel species and P. endodontalis but less than 2% sequence difference between the novel species and P. asaccharolytica. Subsequent DNA-DNA hybridization studies documented that the novel organism was indeed distinct from P. asaccharolytica. We propose the name Porphyromonas uenonis for the novel species. We have recovered P. uenonis from four clinical infections in adults, all likely of intestinal origin, and from the feces of six children. PMID- 15528729 TI - The cag pathogenicity island of Helicobacter pylori is disrupted in the majority of patient isolates from different human populations. AB - The cag pathogenicity island (cag-PAI) is one of the major virulence determinants of Helicobacter pylori. The chromosomal integrity of this island or the lack thereof is speculated to play an important role in the progress of the gastroduodenal pathology caused by H. pylori. We determined the integrity of the cag-PAI by using specific flanking and internally anchored PCR primers to know the biogeographical distribution of strains carrying fully integral cag-PAI with proinflammatory behavior in vivo. Genotypes based on eight selected loci were studied in 335 isolates obtained from eight different geographic regions. The cag PAI appeared to be disrupted in the majority of patient isolates throughout the world. Conservation of cag-PAI was highest in Japanese isolates (57.1%). However, only 18.6% of the Peruvian and 12% of the Indian isolates carried an intact cag PAI. The integrity of cag-PAI in European and African strains was minimal. All 10 strains from Costa Rica had rearrangements. Overall, a majority of the strains of East Asian ancestry were found to have intact cag-PAI compared to strains of other descent. We also found that the cagE and cagT genes were less often rearranged (18%) than the cagA gene (27%). We attempted to relate cag-PAI rearrangement patterns to disease outcome. Deletion frequencies of cagA, cagE, and cagT genes were higher in benign cases than in isolates from severe ulcers and gastric cancer. Conversely, the cagA promoter and the left end of the cag-PAI were frequently rearranged or deleted in isolates linked to severe pathology. Analysis of the cag-PAI genotypes with a different biogeoclimatic history will contribute to our understanding of the pathogen-host interaction in health and disease. PMID- 15528730 TI - Mapping of antigenic sites on the nucleocapsid protein of the severe acute respiratory syndrome coronavirus. AB - Antigenic sites on the nucleocapsid (N) protein of severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) were mapped by Pepscan analysis with overlapping peptides that span the N protein sequence. Two major immunodominant epitopes located in the C-terminal region (amino acids [aa] 362 to 412) and middle region (aa 153 to 178) reacted with more than 75% of sera from SARS patients. Several minor immunodominant epitopes were reactive with about 50% of the SARS sera. Antisera from mice immunized with inactivated SARS-CoV recognized the two major immunodominant epitopes and one antigenic site located adjacent to the N-terminal region (aa 76 to 101), which did not react with the sera from SARS patients. Several monoclonal antibodies against SARS-CoV bound to the N- or C terminal antigenic sites. These results suggest that the above antigenic sites on the N protein are important in eliciting humoral immune response against SARS-CoV in humans and animals and can be used as antigens for developing diagnostic tests. PMID- 15528731 TI - Phaeohyphomycotic soft tissue infections caused by the coelomycetous fungus Microsphaeropsis arundinis. AB - Microsphaeropsis arundinis is an anamorphic fungal plant inhabitant belonging to the form class Coelomycetes. We describe two cases of M. arundinis soft tissue infections in immunosuppressed patients. This organism has not previously been described as causing disease in humans. It was identified on the basis of its typical ostiolate pycnidial conidiomata, ampulliform conidiogenous cells, and small, smooth-walled, brown, cylindrical conidia. PMID- 15528732 TI - Isolation of Bartonella schoenbuchensis from Lipoptena cervi, a blood-sucking arthropod causing deer ked dermatitis. AB - Bartonella schoenbuchensis, which commonly causes bacteremia in ruminants, was isolated from the deer ked Lipoptena cervi and was shown to localize to the midgut of this blood-sucking arthropod, causing deer ked dermatitis in humans. The role of B. schoenbuchensis in the etiology of deer ked dermatitis should be further investigated. PMID- 15528733 TI - Prevalence and characterization of leukotoxin-producing Staphylococcus intermedius in Isolates from dogs and pigeons. AB - Staphylococcus intermedius isolates from dogs (n = 44) and pigeons (n = 62) were categorized into 12 types by intergenic ribosomal DNA spacer polymorphism analysis. All isolates from pigeons were lukS positive and all isolates from dogs were lukS and lukF positive by dot blot analysis. The mean leukotoxicity titer for dog isolates was at least 129-fold higher than that for pigeon isolates. PMID- 15528734 TI - Molecular detection of linezolid resistance in Enterococcus faecium and Enterococcus faecalis by use of 5' nuclease real-time PCR compared to a modified classical approach. AB - A nucleotide transversion from guanine to uracil in the 23S rRNA confers linezolid resistance. We describe a real-time PCR using two Taqman probes that detects a single mutated allele among the genomes of Enterococcus faecium and Enterococcus faecalis. Results were confirmed by a classical approach involving LabChip technology assayed with an Agilent Bioanalyzer 2100. PMID- 15528735 TI - Convenience of serum for visceral leishmaniasis diagnosis by PCR. AB - In this retrospective study, the usefulness of a PCR performed on serum for primary diagnosis and monitoring of Mediterranean visceral leishmaniasis (MVL) was assessed. In the case of primary diagnosis of MVL, the serum PCR showed a sensitivity of 97% and a specificity of 95%, with positive and negative predictive values of 94 and 97%, respectively. PMID- 15528736 TI - Utility of molecular identification in opportunistic mycotic infections: a case of cutaneous Alternaria infectoria infection in a cardiac transplant recipient. AB - We report on a case of cutaneous infection caused by Alternaria infectoria in a cardiac transplant recipient. A rapid molecular diagnosis was obtained by sequence analysis of the internal transcribed spacer domain of the 5.8S ribosomal DNA region amplified from colonies developed on Sabouraud medium. Treatment consisted of a combination of systemic antifungal therapy, first with amphotericin B and then with itraconazole. PMID- 15528737 TI - Complexity of Klebsiella pneumoniae isolates resistant to both cephamycins and extended-spectrum cephalosporins at a teaching hospital in Taiwan. AB - Among 99 clinical Klebsiella pneumoniae isolates resistant to cefoxitin and extended-spectrum cephalosporins, coexistence of AmpC (DHA-1, CMY-2, or CMY-8) and extended-spectrum beta-lactamases (CTX-M and/or SHV) was detected in a total of 35. The remainder produced AmpC (n = 42), extended-spectrum beta-lactamases (n = 9), metallo-beta-lactamases (n = 2), or none of these enzymes (n = 11). Phenotypic characteristics of these isolates were demonstrated. PMID- 15528738 TI - Evaluation of a modified gen-probe amplified direct test for detection of Mycobacterium tuberculosis complex organisms in cerebrospinal fluid. AB - Laboratory evidence for tuberculous meningitis is difficult to acquire due to the low numbers of organisms present in cerebrospinal fluid (CSF) and the presence of nucleic acid amplification inhibitors. The Amplified Mycobacterium tuberculosis Direct Test (MTD) is sensitive and specific for the direct detection of M. tuberculosis complex in respiratory samples but has not been approved for CSF. We evaluated a modified version of the current MTD, optimized for use with CSF samples. Samples were prepared by spiking CSF with various numbers of M. tuberculosis complex organisms. The modified MTD performance was compared with results obtained using a purified RNA sample extracted using the Qiagen RNeasy Protect Bacteria Mini Kit. By use of CSF artificially spiked with M. tuberculosis complex, the sensitivity of the modified MTD was 100% (six of six) for CSF samples containing approximately 600 CFU/ml, 78% (seven of nine) for approximately 60 CFU/ml, 50% (three of six) for 6 CFU/ml, and 17% (one of six) for samples with <1 CFU/ml. The specificity of the modified MTD method was 100% (22 of 22). The sensitivity of the Qiagen MTD method was 100% for CSF samples containing approximately 600 CFU/ml (six of six) and approximately 60 CFU/ml (nine of nine), 50% for samples with approximately 6 CFU/ml (three of six), and 50% for samples with <1 CFU/ml (three of six). The specificity of the Qiagen MTD method was 86% (19 of 22). With the Qiagen MTD method, however, initial results were equivocal for 14 of the 27 (52%) positive samples, requiring repeat analysis, whereas with the modified MTD, only 1 of 27 (4%) was equivocal. The modified MTD for CSF samples was less time-consuming and less expensive and resulted in considerably fewer equivocal results than the Qiagen MTD method did. PMID- 15528739 TI - Mycobacterium avium subsp. paratuberculosis strains isolated from Crohn's disease patients and animal species exhibit similar polymorphic locus patterns. AB - Analysis of short sequence repeats of Mycobacterium avium subsp. paratuberculosis isolated from Crohn's disease patients identified two alleles, both of which clustered with strains derived from animals with Johne's disease. Identification of a limited number of genotypes among human strains implies the existence of human disease-associated genotypes and strain sharing with animals. PMID- 15528740 TI - Sensitive and specific detection of Yersinia pseudotuberculosis by loop-mediated isothermal amplification. AB - We developed a loop-mediated isothermal amplification method able to detect Yersinia pseudotuberculosis strains in 30 min by using six primers designed by targeting the inv gene. This method is more sensitive than PCR and might be a useful tool for detecting and identifying Y. pseudotuberculosis. PMID- 15528741 TI - Monitoring for presence of potentially xenotic viruses in recipients of pig islet xenotransplantation. AB - This study represents a long-term follow-up of human patients receiving pig islet xenotransplantation. Eighteen patients had been monitored for up to 9 years for potentially xenotic pig viruses: pig endogenous retrovirus, pig cytomegalovirus, pig lymphotropic herpesvirus, and pig circovirus type 2. No evidence of viral infection was found. PMID- 15528742 TI - Fibronectin-binding protein gene recombination and horizontal transfer between group A and G streptococci. AB - We report evidence of interspecies gene transfer between the important virulence factor genes sfbI and gfbA. Because the identified group G streptococcus gfbA types possess DNA cassettes that can be identified in a number of group A streptococcus strains, it appears that homologous recombination is occurring between these species. PMID- 15528743 TI - False-positive Aspergillus galactomannan enzyme-linked immunosorbent assay results in vivo during amoxicillin-clavulanic acid treatment. AB - Positive Platelia Aspergillus test results were observed in consecutive serum samples from an immunocompromised host during amoxicillin-clavulanic acid treatment, and a correlation between plasmatic amoxicillin concentration and galactomannan optical density index was observed. Amoxicillin-clavulanic acid vials tested positive for galactomannan but were negative for Aspergillus DNA. PMID- 15528744 TI - Polymorphisms of the fimbria fim3 gene of Bordetella pertussis strains isolated in Canada. AB - The fim genes which code for the fimbria protective antigens present in both the inactivated whole-cell and acellular vaccines were analyzed in 86 Canadian Bordetella pertussis isolates. At least one of the novel mutations identified was found to involve a surface epitope that has been mapped by serum antibodies from infected or vaccinated subjects. PMID- 15528745 TI - Identification and phylogeny of Enterobacter sakazakii relative to Enterobacter and Citrobacter Species. AB - The phylogenetic relationships of Enterobacter sakazakii strains were investigated using 16S ribosomal DNA (rDNA) and hsp60 sequencing. Each analysis distributed E. sakazakii strains among four clusters, indicating substantial taxonomic heterogeneity. The E. sakazakii type strain 16S rDNA sequence was 97.8% similar to that of Citrobacter koseri but 97.0% similar to that of Enterobacter cloacae. PMID- 15528747 TI - Characterization of a West Nile virus isolate from a human on the Gulf Coast of Texas. AB - Genetic characterization of a human cerebrospinal fluid West Nile virus isolate from Beaumont, Texas, revealed several nucleotide changes and amino acid substitutions that differentiated it from all other North American strains isolated to date, suggesting that isolates from the Texas Gulf Coast may form a unique genetic group among North American strains. PMID- 15528746 TI - Prevalence of hepatitis E virus (HEV) Infection in wild boars and deer and genetic identification of a genotype 3 HEV from a boar in Japan. AB - Zoonotic transmission of hepatitis E virus (HEV) from captured wild deer or boars to humans has been suggested. Antibody to HEV was detected in 9% of 35 wild boars and 2% of 117 wild deer tested, and a presumably indigenous HEV of genotype 3 was isolated from a boar in Japan. PMID- 15528748 TI - Clonal spread of IMP-1-producing Pseudomonas aeruginosa in two hospitals in Singapore. AB - Thirty-six isolates of carbapenem-resistant Pseudomonas aeruginosa were studied. Pulsed-field gel electrophoresis revealed the presence of two clones. One clone carried a bla(IMP-1) gene identical to that first described in Japan. The other clone carried a bla(IMP-1) variant containing four silent mutations. One isolate with a unique pulsed-field gel electrophoresis pattern contained bla(IMP-7). PMID- 15528749 TI - Stochastic transmission of multiple genotypically distinct Anaplasma marginale strains in a herd with high prevalence of Anaplasma infection. AB - Multiple genotypically unique strains of the tick-borne pathogen Anaplasma marginale occur and are transmitted within regions where the organism is endemic. In this study, we tested the hypothesis that specific A. marginale strains are preferentially transmitted. The study herd of cattle (n = 261) had an infection prevalence of 29% as determined by competitive inhibition enzyme-linked immunosorbent assay and PCR, with complete concordance between results of the two assays. Genotyping revealed the presence of 11 unique strains within the herd. Although the majority of the individuals (70 of 75) were infected with only a single A. marginale strain, five animals each carried two strains with markedly distinct genotypes, indicating that superinfection does occur with distinct A. marginale strains, as has been reported with A. marginale and A. marginale subsp. centrale strains. Identification of strains in animals born into and infected within the herd during the period from 1998 to 2003 revealed no significant difference from the overall strain prevalence in the herd, results that do not support the occurrence of preferential strain transmission within a population of persistently infected animals and are most consistent with pathogen strain transmission being stochastic. PMID- 15528750 TI - Effect of transport time, temperature, and concentration on the survival of group B streptococci in amies transport medium. AB - The impact of transport time and temperature on survival of group B streptococci (GBS) in Amies transport medium was evaluated. Viability of 10 or more CFU of GBS was maintained for 4 days at 24 or 3 degrees C. However, there was a significant decrease in viability for GBS held at 30 degrees C for 4 days. PMID- 15528752 TI - Comparison of the sensitivities and viral load values of the AMPLICOR HIV-1 MONITOR version 1.0 and 1.5 tests. AB - This study compared the sensitivity and viral load values of the AMPLICOR HIV-1 MONITOR microwell version 1.0, microwell version 1.5, and COBAS version 1.5 tests. Based on the percentage of positive replicates, the microwell version 1.5 and COBAS version 1.5 tests are more sensitive than the microwell version 1.0 test. Viral load values obtained with the COBAS version 1.5 test are lower than those obtained with either the microwell version 1.0 or microwell version 1.5 test. PMID- 15528751 TI - Spoligotyping profile change caused by deletion of a direct variable repeat in a Mycobacterium tuberculosis isogenic laboratory strain. AB - Spoligotyping is a major tool for molecular typing of Mycobacterium tuberculosis complex organisms. For epidemiological purposes, strains are considered clonal only when their spoligotyping patterns are identical. We report a change in the spoligotyping profiles of truly isogenic strains (a clinical isolate and a subculture derived in the laboratory) caused by deletion of a direct variable repeat. Without the information about the relationship between them, a link between these strains would have gone unnoticed. Evolutionary events should be taken into account in the interpretation of spoligotyping results and in the design of databases. PMID- 15528753 TI - Association between a high BK virus load in urine samples of patients with graft versus-host disease and development of hemorrhagic cystitis after hematopoietic stem cell transplantation. AB - BK virus (BKV) load in urine alone or in combination with acute graft-versus-host disease (GVHD) was correlated to development of hemorrhagic cystitis (HC). BKV load in combination with acute GVHD discriminated the best, while BKV and viral load alone, but not GVHD, still showed predictive ability for HC. PMID- 15528755 TI - Differentiation of Rhizomucor species on the basis of their different sensitivities to lovastatin. AB - The opportunistic pathogens Rhizomucor pusillus and Rhizomucor miehei may be agents of frequently fatal mycotic diseases. In the present study, the susceptibilities of 27 clinical and environmental isolates of R. miehei and R. pusillus to lovastatin under different culturing conditions were investigated. Most of the R. miehei strains grew at lovastatin concentrations as high as 64 to 128 microg/ml. In contrast, the inhibitory effect of lovastatin on all of the R. pusillus strains was evident at lovastatin concentrations as low as 1 to 2 microg/ml. A simple and reliable method for species-level differentiation, based on the significantly higher sensitivity of R. pusillus to lovastatin than that of R. miehei, was elaborated. According this, on malt extract agar containing 6 mug of lovastatin/ml, R. pusillus is not able to produce colonies, while R. miehei will form compact colonies. PMID- 15528754 TI - Demonstration of two distinct hemotropic mycoplasmas in French dogs. AB - In North America it has been shown that distinct hemotropic mycoplasmas exist in dogs. Blood samples from 460 French dogs were analyzed by PCR to evaluate hemoplasma infection status. Seventy-one dogs (15.4%) were positive; of these, 44 (9.6%) were infected with an organism closely related to "Candidatus Mycoplasma haemoparvum" only, 15 (3.3%) were infected with Mycoplasma haemocanis only, and 12 dogs (2.6%) were dually infected with both organisms. PMID- 15528756 TI - An unexpected experimental pitfall in the molecular diagnosis of bacterial endophthalmitis. AB - General primer-mediated ribosomal DNA amplification during endophthalmitis may improve the quality of diagnostic microbiology. However, extreme care needs to be taken not to introduce contaminating bacterial DNA during surgery procedures. The use of decontaminating iodine solutions can lead to such contamination due to the presence of DNA from Pseudomonas-like organisms. PMID- 15528757 TI - Natural Clostridium botulinum type C toxicosis in a group of cats. AB - Clinical signs of botulism were observed in a group of eight cats, four of which died, after being fed pelican carrion. Clostridium botulinum type C was isolated from one cat. The microorganism and its toxin were found in the pelican. This is apparently the first report of natural botulism in cats. PMID- 15528758 TI - Laboratory-confirmed transmission of vaccinia virus infection through sexual contact with a military vaccinee. AB - A laboratory-confirmed, inadvertent transmission of vaccinia virus from an unusual source highlights the importance of epidemiologic tracing, proper biosafety practices in the clinical diagnostic laboratories, and educating clinicians and laboratorians to potential bioterrorism-initiated outbreaks as well as look-alike disease discrimination. PMID- 15528759 TI - Disseminated Beauveria bassiana infection in a patient with acute lymphoblastic leukemia. AB - We describe a case of disseminated Beauveria bassiana infection in a patient with acute lymphoblastic leukemia. Her infection was successfully treated with amphotericin B and itraconazole. B. bassiana is rarely reported as a human pathogen. It is commonly found in soil and because of its pathogenicity to many insect species is incorporated into several pesticides. PMID- 15528760 TI - Interstitial pneumonitis and coinfection of human herpesvirus 6 and Pneumocystis carinii in a patient with hypogammaglobulinemia. AB - Human herpesvirus 6 (HHV-6) has occasionally been associated with cases of interstitial pneumonitis, mainly in individuals with impaired cellular immunity. Here we report for the first time severe interstitial pneumonitis with simultaneous HHV-6 and Pneumocystis carinii infections in the lung tissue of a young patient with hypogammaglobulinemia. PMID- 15528761 TI - Fatal cerebral phaeohyphomycosis due to Curvularia lunata in an immunocompetent patient. AB - Curvularia infections in humans are relatively uncommon despite the ubiquitous presence of this soil-dwelling dematiaceous fungus in the environment. Originally thought to be solely a pathogen of plants, Curvularia has been described as a pathogen of humans and animals in the last half-century, causing respiratory tract, cutaneous, and corneal infections. Only three previous cases of central nervous system involvement by Curvularia have been documented in the medical literature. We report a fatal case of cerebral Curvularia infection in which there was no known history of immunocompromise or prior respiratory tract or sinus infection in the patient. PMID- 15528762 TI - Genetic confirmation of quinine-resistant Plasmodium falciparum malaria followed by postmalaria neurological syndrome in a traveler from Mozambique. AB - A case of quinine-resistant Plasmodium falciparum malaria, followed by a postmalaria neurological syndrome and a recurrence episode, is described. Genetic characterization of the P. falciparum isolate obtained by analysis of msp1 and msp2 amplicons revealed the coexistence of two genotypes causing the first malaria episode and the presence of a unique isolate responsible for the recurrence. PMID- 15528763 TI - Aeromonas popoffii urinary tract infection. AB - Aeromonas popoffii is a recently described species isolated mainly from freshwater. An isolate of Aeromonas popoffii was found to be responsible for a urinary tract infection in a 13-year-old boy suffering from spina bifida with enterocystoplasty. This is the first reported case of human infection attributed to this species. PMID- 15528764 TI - Catheter-related bloodstream infection caused by Mycobacterium brumae. AB - Mycobacterium brumae is a rapidly growing environmental mycobacterial species identified in 1993; so far, no infections by this organism have been reported. Here we present a catheter-related M. brumae bloodstream infection in a 54-year old woman with breast cancer. The patient presented with high fever (39.7 degrees C), and >1,000 colonies of M. brumae grew from a quantitative culture of blood drawn through the catheter. A paired peripheral blood culture was negative, however, suggesting circulational control of the infection. The patient was treated empirically with meropenem and vancomycin, and the fever resolved within 24 h. The catheter was removed a week later, and from the tip M. brumae was isolated a second time, suggesting catheter colonization. The organism was identified by colonial morphology, sequence analysis of the 16S rRNA gene, and biochemical tests. PMID- 15528765 TI - Seroprevalence of Mycobacterium paratuberculosis in patients with Crohn's Disease. PMID- 15528766 TI - The viable but nonculturable concept, bacteria in urine samples, and Occam's razor. PMID- 15528767 TI - ASM's role in writing practice guidelines. PMID- 15528768 TI - Caloric restriction and cardiovascular aging in cynomolgus monkeys (Macaca fascicularis): metabolic, physiologic, and atherosclerotic measures from a 4-year intervention trial. AB - Caloric restriction (CR) retards aging processes, extends maximal life span, and consistently improves insulin resistance in lower species. Insulin resistance is associated with cardiovascular disease, but data is lacking demonstrating that increased insulin sensitivity reduces atherosclerosis progression. We initiated a study in 32 adult cynomolgus monkeys to evaluate increased insulin sensitivity secondary to CR on atherosclerosis extent. Following pretrial determinations, animals were randomized to a moderately atherogenic (0.25 mg cholesterol/Cal containing 30% of calories from fat)-fed control group or CR group (30% reduction) with equivalent dietary cholesterol intake. CR significantly improved insulin sensitivity and reduced intraabdominal fat over the 4-year intervention, while no significant differences were seen for the lipid profile between groups. Despite improved insulin sensitivity with CR, atherosclerosis extent did not differ between the ad libitum-fed or CR groups. These studies demonstrate that CR significantly improves insulin sensitivity, but when elevated plasma cholesterol concentrations were held similar, there was no effect on atherosclerosis extent. However, the composition of these lesions and changes in endothelial function may have been improved but were not evaluated in this study. Thus, further studies are needed to determine if improved insulin sensitivity might decrease arterial inflammation and improve endothelial function, despite no changes in atherosclerosis extent. PMID- 15528769 TI - The effects of aging and treadmill running on soleus and gastrocnemius muscle morphology in the senescence-accelerated mouse (SAMP1). AB - We investigated the effects of aging on the soleus and gastrocnemius muscles in male SAMP1 (senescence-accelerated mouse prone 1). Body mass, muscle wet weight, fiber size, and the percent of type II fibers declined from 50 weeks of age. Voluntary motor behavior also significantly declined with age. Furthermore, we examined the effects of high (twice daily) and low (once daily) frequency treadmill running, for 6 weeks at 5 days per week, beginning when the mice were 50 weeks old. Muscle fiber size for the high frequency running significantly increased. Pathological fiber alterations in these mice were increased by running, especially by high frequency running. This suggests that age-related muscle morphological changes in SAMP1 occurs from 50 weeks of age, and that the decline in voluntary motor behavior is an important factor in aging muscle atrophy. In addition, high frequency running is more beneficial for aged muscle hypertrophy. This model is useful for studying the acceleration of the aging process in skeletal muscle of the SAM. PMID- 15528770 TI - Genetic loci that influence cause of death in a heterogeneous mouse stock. AB - A genome scan was conducted to seek evidence for polymorphic genes that influence cause of death in mice produced by a cross between CB6F1 females and C3D2F1 males. Loci on chromosomes 1 and 4 were found to modulate risk of lymphoma. A locus on chromosome 4 influenced risk of mammary adenocarcinoma among multiparous female mice, but had no significant effect in virgin females. A chromosome 4 locus was found to modulate risk of death from either hemangiosarcoma or fibrosarcoma. A suggestive linkage was noted (at p =.09) between a marker on chromosome 11 and hepatocellular carcinoma. Lastly, a locus on chromosome 6 was noted to influence the likelihood that pulmonary adenocarcinoma would be present at death. The collection of normal and neoplastic tissues from 1004 terminal necropsies, together with genetic information, provides a valuable resource for further studies of the genetic influences on late-life diseases in mice. PMID- 15528771 TI - Expression and significance of integrin-linked kinase in cultured cells, normal tissue, and diseased tissue of aging rat kidneys. AB - Integrin-linked kinase (ILK) is an integrin-binding cytoplasmic protein that has been implicated in regulating numerous cellular processes and fibronectin (Fn) deposition through mediated integrin, but the expression and significance of ILK in the aging kidney have not yet been reported. We report here that mRNA and protein expression of ILK increased in primary cultured mesangial and tubular epithelial cells, and normal and unilateral ureteral obstructed kidney tissues in 28-month-old rats but not in 3-month-old rats, moreover, accompanied by the over expression of Fn and integrin-beta1 in the aging kidney, by means of Northern blot, Western blot, and immunofluorescent double-staining immunohistochemistry. In addition, in the primary cultured kidney cells, ILK expression was positively correlated with senescence-associated beta-gal positive staining and negatively correlated with cellular proliferation. The results suggest that ILK may be involved in the fibrotic or senescent process in the aging kidney. PMID- 15528773 TI - A step back in time: is there a place for older drugs in the treatment of dementia? PMID- 15528772 TI - Modulation of PPAR in aging, inflammation, and calorie restriction. AB - Peroxisome proliferator-activated receptors (PPARs), members of the nuclear hormone receptor superfamily of transcription factors, are key regulators in various pathophysiological processes related to energy metabolism including lipid, carbohydrate metabolism, and inflammation. At present, little information is on the effect of age and calorie restriction (CR) on PPARs. In the present study, we investigated how age and CR (60% of the ad libitum intake) modulate PPARs in kidneys obtained from Fischer 344 rats, ages 13 and 25 months. Results showed that nuclear protein, mRNA level, and DNA binding activity of PPARs decreased with age, while CR blunted the reduction. Our findings were verified in separate experiments in which rats were injected with lipopolysaccharide, with the result of increased susceptibility to inflammation. Based on these findings, we conclude that the altered expression of PPARs may be due to increased oxidative stress with age, and that CR prevents these decreases through its antioxidative action. PMID- 15528774 TI - Perceptions of geriatric medicine junior faculty on success in academic medicine: the Saint Louis University Geriatric Academy (SLUGA) Faculty Development Program. PMID- 15528775 TI - Teaching physicians geriatric principles: a randomized control trial on academic detailing plus printed materials versus printed materials only. AB - BACKGROUND: We compared the effectiveness of academic detailing with printed materials, versus printed materials only, on promoting geriatric knowledge among physicians. METHODS: 31 physicians were randomly assigned to receive academic detailing plus printed materials (group 1 intervention, n = 16), or printed materials alone (group 2 control, n = 15), on 5 geriatric topics identified from a needs assessment survey. Two participants withdrew from group 2, leaving 13 in the control group. Both groups received printed educational materials between baseline and time 1, and at time 2. Only group 1 received an additional 15 minute, one-on-one education session with a geriatrician at time 2. The primary endpoint was knowledge retention, measured by the numeric score of a 5-item questionnaire (range 0 to 5), comprised of items from the Geriatrics Knowledge Test. Knowledge retention was measured at baseline, 1 week (time 1), and on average 29 weeks later (time 2). RESULTS: Most participants were postgraduate trainees. The mean knowledge score in all participants decreased from 3.6 +/- 1.2 at baseline to 3.1 +/- 1.2 at time 1 (p =.006). 19 participants (9 in group 1 and 10 in group 2) completed the knowledge questionnaire at time 2. At baseline, group 1 scored lower than group 2 (3.4 +/- 1.3 versus 3.8 +/- 1.1, p =.39); whereas at time 2, group 1 scored significantly higher than group 2 (4.7 +/- 0.7 versus 3.9 +/- 0.7, p =.034). Academic detailing plus printed materials produced higher mean score change from baseline (1.1 +/- 1.3) than printed materials alone (0.0 +/- 1.1, p =.053). CONCLUSIONS: Academic detailing plus printed materials improved knowledge retention among physicians, whereas printed materials only did not. PMID- 15528776 TI - Effectiveness of nutritional supplements on cognitive functioning in elderly persons: a systematic review. AB - BACKGROUND: The effectiveness of nutritional supplementation in improving cognitive functioning is evaluated in elderly people. METHODS: The authors systematically reviewed randomized controlled trials that compared nutritional supplementation with a placebo treatment. Trials were identified from a MEDLINE search and from reference lists of identified studies and review articles. From each trial, information was gathered on the number and age of persons studied; the type, dosage, and duration of the intervention; and the assessed outcome measures. RESULTS: From 1086 titles, 571 articles were excluded based on their titles. Of the remaining 467 articles, the abstracts were read and 422 articles were excluded based on information found there. The remaining articles were screened for quality aspects of the study design, leaving 21 proper randomized, controlled trials. These trials are discussed in three groups according to the type of supplementation: multinutrient intervention or single components with or without a putative mechanism. Twelve studies, which were evenly distributed among the three supplement groups, found significantly positive effects of nutritional intervention on cognitive functioning, whereas nine studies did not. None of the studies found a significantly negative effect of nutritional intervention. CONCLUSIONS: Shortcomings in methodology varying from the duration of intervention to outcome measures partly explain discrepancies in findings. Despite the heterogeneity in trial design, the results of this review suggest that nutritional supplements may improve the cognitive functioning of elderly persons and do no harm. Further well-designed studies are needed to support these findings. PMID- 15528777 TI - The timing of activity rhythms in patients with dementia is related to survival. AB - BACKGROUND: Older adults with dementia often have disruptions in circadian rhythms, including disruptions of the rest-activity rhythm. These disruptions are a product of internal neuronal activity and external environmental influences, both of which are deficient in dementia. However, the consequences of disturbed rhythms are unknown. This study examined the relationship between rest-activity rhythms and death in patients with dementia. METHODS: The authors recruited 149 older adults with dementia (104 women; mean age, 84.1 years) from nursing homes. Activity was recorded with wrist actigraphs from each participant for 3 days. Survival was determined by examining public death records. Cox proportional hazards models were used to determine which aspects of rest-activity rhythms were related to survival. RESULTS: The timing of each participant's rest-activity rhythm compared with a sample of persons without dementia was related to survival, such that those who more closely resembled the persons without dementia lived longer. CONCLUSIONS: Although rest-activity rhythms as a whole were not related to survival, the timing of the rhythm was. Patients with dementia appear to develop an abnormal timing of their rhythms, which is predictive of shorter survival. It may be possible to intervene with these patients to correct the timing of their rhythms and possibly prolong their lives. PMID- 15528778 TI - Refining the categorization of physical functional status: the added value of combining self-reported and performance-based measures. AB - BACKGROUND: When considered individually, self-reported functional status and performance-based functional status predict functional status decline and mortality. However, what additional prognostic information is gained by combining these approaches remains unknown. METHODS: The authors used three waves of three sites (5138 participants) of the Established Populations for Epidemiologic Studies of the Elderly to determine the prognostic value of individual and combined approaches. Baseline self-reported (mobility and activities of daily living [ADL] items) and performance-based (Physical Performance Score) functional status information was classified into three and four hierarchical categories, respectively. RESULTS: Based on self-reported information alone, at 1 year, 73% participants had not changed, 15% declined, 6% improved, and 6% died. At 4 years, 53% had not changed, 24% declined, 2% improved, and 22% died. Based on performance-based assessment alone, at 4 years, 33% of the sample remained stable, 37% declined, 6% improved, and 24% died. In the top two self-reported categories, functioning on the performance-based assessment varied widely. Among those who were independent in all self-reported functioning, approximately 40% scored in each of the top two performance-based categories. Among persons in the top two self-reported categories, poorer performance was associated with progressively higher 1-year and 4-year mortality rates. Among persons with impaired mobility and at least 1 ADL dependency, the mortality rate was high and was not influenced by performance-based score. CONCLUSIONS: Combining self reported and performance-based measurements can refine prognostic information, particularly among older persons with high self-reported functioning. However, if ADL dependency is present, performance-based measures do not add prognostic value regarding mortality. PMID- 15528779 TI - Low-intensity exercise and reduction of the risk for falls among at-risk elders. AB - BACKGROUND: Among elderly persons, falls account for 87% of all fractures and are contributing factors in many nursing home admissions. This study evaluated the effect of an easily implemented, low-intensity exercise program on the incidence of falls and the time to first fall among a clinically defined population of elderly men and women. METHODS: This community-based, randomized trial compared the exercise intervention with a no-intervention control. The participants were 294 men and women, aged 60 years or older, who had either a hospital admission or bed rest for 2 days or more within the previous month. Exercise participants were scheduled to attend exercise sessions lasting 45 minutes, including warm-up and cool-down, 3 times a week for 8 weeks (24 sessions). Assessments included gait and balance measures, self-reported physical function, the number of medications being taking at baseline, participant age, sex, and history of falling. Falls were tracked for 1 year after each participant's baseline assessment. RESULTS: 29% of the study participants reported a fall during the study period. The effect of exercise in preventing falls varied significantly by baseline physical function level (p < or =.002). The risk for falls decreased for exercise participants with low baseline physical functioning (hazard ratio,.51) but increased for exercise participants with high baseline physical functioning (hazard ratio, 3.51). CONCLUSIONS: This easily implemented, low-intensity exercise program appears to reduce the risk for falls among elderly men and women recovering from recent hospitalizations, bed rest, or both who have low levels of physical functioning. PMID- 15528781 TI - Children of Alzheimer patients: more data needed. PMID- 15528780 TI - Measuring medical burden using CIRS in older veterans enrolled in UPBEAT, a psychogeriatric treatment program: a pilot study. AB - BACKGROUND: A quantitative measure of medical burden is needed to assess medical comorbidities in psychogeriatric patients. The Cumulative Illness Rating Scale (CIRS) is the most widely used instrument for measuring medical burden in psychogeriatric research. Many clinicians, however, are discouraged by the requirement to project the persistence of acute conditions and therefore do not use the scale. The goal of this pilot study was to determine whether the inclusion of acute medical conditions undermines the usefulness of the CIRS. No such comparison was found in the existing literature. METHODS: Included in this study were 95 patients previously enrolled in the Unified Psychogeriatric Biopsychosocial Evaluation and Treatment (UPBEAT) demonstration program. All were male veterans of the U.S. armed forces who were admitted to acute medical or surgical inpatient units and who had positive screening results for anxiety, depression, or alcohol abuse. Two types of retrospective CIRS ratings were made for each patient: one included (CIRS-IP) and the other excluded (CIRS-PH) acute conditions. For each type of rating (CIRS-IP and CIRS-PH), 7 CIRS scores were computed according to methods reported in the literature. Survival time during 24 months of follow-up was used as a measure of health outcome indicating medical burden. RESULTS: With 1 exception, CIRS-IP and corresponding CIRS-PH scores were highly correlated (.70 < r <.99; p <.001). And, for 5 of 7 scores, both CIRS-IP and CIRS-PH were significantly associated with survival time (p <.05). CONCLUSIONS: Results suggest that the CIRS can be used as an indicator of medical burden even with the inclusion of acute conditions. If replicated, these findings may increase CIRS use and thus aid the effort to encourage clinicians working with psychogeriatric patients to use standardized instruments to document medical burden. PMID- 15528782 TI - Advancing in the analysis of disability in cognitively impaired older adults. PMID- 15528783 TI - Lynch syndrome (HNPCC) and microsatellite instability. PMID- 15528784 TI - Lynch syndrome: history and current status. PMID- 15528785 TI - Colorectal carcinogenesis: MSI-H versus MSI-L. AB - Microsatellite instability (MSI) is a well-recognized phenomenon that is classically a feature of tumors in the hereditary non-polyposis colorectal syndrome. Ten to 15% of sporadic colorectal cancers, however, will have MSI. Microsatellite unstable tumors can be divided into two distinct MSI phenotypes: MSI-high (MSI-H) and MSI-low (MSI-L). MSI sporadic colorectal cancers with a high level of MSI (MSI-H) form a well defined group with distinct clinicopathologic features characterized by an overall better long-term prognosis. These sporadic MSI-H colorectal tumors most often arise from the epigenetic silencing of the mismatch repair gene MLH1. In contrast, MSI-L colorectal tumors have not been shown to differ in their clinicopathologic features or in most molecular features from microsatellite stable (MSS) tumors. Unlike MSI-H tumors, MSI-L tumors appear to arise through the chromosomal instability carcinogenesis pathway, similar to MSS tumors. Some groups have reported more frequent mutations in K-ras and in the methylation of methylguanine transferase in MSI-L tumors, but others have questioned these findings. Therefore, although the use of the MSI-L category is widespread, there continues to be some debate as to whether a discrete MSI-L group truly exists. Rather, it has been suggested that MSI-L tumors differ quantitatively from MSS tumors but do not differ qualitatively. Future studies will need to evaluate the specific mutations in non-MSI-H tumors in an attempt to sub-classify MSI-L tumors with regard to MSS tumors so that subtle differences between these two sub-groups can be identified. PMID- 15528786 TI - Identification of HNPCC by molecular analysis of colorectal and endometrial tumors. AB - Hereditary nonpolyposis colorectal cancer (HNPCC, Lynch syndrome) is a dominantly inherited syndrome characterized by the development of colorectal cancer, endometrial cancer and other cancers and the presence of microsatellite instability (MSI) in tumors. The Bethesda guidelines have been proposed for the identification of families suspected of HNPCC that require further molecular analysis. We have evaluated the yield of MSI-analysis in a large series of Dutch families suspected of HNPCC. We also analysed whether the loss of mismatch repair (MMR) protein detected by immunohistochemistry (IHC) of colorectal cancer (CRC) and endometrial cancer correlated with the presence of MSI and/or a MMR gene mutation. The results showed that the Bethesda criteria with a few modifications are appropriate to identify families eligible for genetic testing. In addition, we found that MSI and IHC-analysis of CRC using antibodies against MLH1, MSH2, MSH6 and PMS2 proteins are equally effective for identifying carriers of the known MMR gene defects. However, as long as the role of other putative MMR genes in hereditary CRC has not been elucidated, IHC-analysis cannot completely replace MSI. For this reason, we prefer MSI-analysis as first step in families suspected of HNPCC. On the other hand, in families fulfilling the revised Amsterdam criteria in which the probability of detecting a mutation is relatively high, we would recommend IHC as first diagnostic step because the result might predict the specific underlying MMR gene mutation. MSI or IHC-analysis of endometrial cancer alone was found to be less sensitive compared with these tests performed in colorectal cancer. Therefore, probably the best approach in the analysis of this cancer is to perform both techniques. The identification of HNPCC is important as it makes it possible to target effective preventative measures. Our studies showed that MSI and IHC analysis of colorectal and endometrial cancer, are reliable cost-effective tools that can be used to identify patients with HNPCC. PMID- 15528787 TI - Role of the pathologist in the diagnosis of hereditary non-polyposis colorectal cancer. AB - The aim of this paper is to indicate how the pathologist may suspect a diagnosis of hereditary non-polyposis colorectal cancer (HNPCC) on the basis of histological criteria and patient age alone. A single morphological feature, namely the presence of intra-epithelial lymphocytes (tumor infiltrating lymphocytes), identifies the majority of colorectal cancers (CRC) with the DNA microsatellite instability-high phenotype. A number of pathological criteria can help to distinguish HNPCC from sporadic MSI-H CRC, though age below 60 years is an important pointer towards HNPCC. Immunohistochemistry to demonstrate loss of expression of DNA mismatch repair genes serves as a highly reliable test of mismatch repair deficiency if antibodies to hMLH1, hMSH2, hMSH6 and hPMS2 are employed. PMID- 15528788 TI - MSI-testing in hereditary non-polyposis colorectal carcinoma (HNPCC). AB - Genomic instability at simple repeated sequences, termed microsatellite instability (MSI), plays an important role in the analysis of sporadic and hereditary colon cancers. In hereditary non-polyposis colorectal cancer syndrome (HNPCC) more than 90% of cases show MSI, whereas only 10-15% of sporadic colorectal cancers do so. Thus, microsatellite analysis is commonly used as the first diagnostic screening test for HNPCC. In 1997, an international collaborative workshop sponsored by the National Cancer Institute (NCI) proposed a set of guidelines for MSI-testing to improve reliability and reproducibility of the analysis as well to allow comparisons between different studies and different laboratories. In this review we assess the value of current protocols for MSI testing and discuss some diagnostic pitfalls. Our findings support continued use of the MSI marker panel recommended in 1997. Additionally, MSI-testing should be improved by use of microdissection, which helps to identify additional patients with MSI due to enrichment of tumor cells and therefore increased sensitivity. In our view, immunohistochemical staining for mismatch repair protein expression is not a substitute for MSI-analysis but complements MSI screening and helps direct further testing. In summary, MSI-analysis is a highly sensitive and reliable screening method for HNPCC, that requires a well-equipped laboratory as well as an experienced pathologist. Integration of family history and histo-pathological features is also critical. PMID- 15528790 TI - Quasimonomorphic mononucleotide repeats for high-level microsatellite instability analysis. AB - Microsatellite instability (MSI) analysis is becoming more and more important to detect sporadic primary tumors of the MSI phenotype as well as in helping to determine Hereditary Non-Polyposis Colorectal Cancer (HNPCC) cases. After some years of conflicting data due to the absence of consensus markers for the MSI phenotype, a meeting held in Bethesda to clarify the situation proposed a set of 5 microsatellites (2 mononucleotide repeats and 3 dinucleotide repeats) to determine MSI tumors. A second Bethesda consensus meeting was held at the end of 2002. It was discussed here that the 1998 microsatellite panel could underestimate high-level MSI tumors and overestimate low-level MSI tumors. Amongst the suggested changes was the exclusive use of mononucleotide repeats in place of dinucleotide repeats. We have already proposed a pentaplex MSI screening test comprising 5 quasimonomorphic mononucleotide repeats. This article compares the advantages of mono or dinucleotide repeats in determining microsatellite instability. PMID- 15528789 TI - Development of a fluorescent multiplex assay for detection of MSI-High tumors. AB - Determining whether a tumor exhibits microsatellite instability (MSI) is useful in identifying patients with hereditary non-polyposis colorectal cancer and sporadic gastrointestinal cancers with defective DNA mismatch repair (MMR). The assessment of MSI status aids in establishing a clinical prognosis and may be predictive of tumor response to chemotherapy. A reference panel of five markers was suggested for MSI analysis by a National Cancer Institute (NCI) workshop in 1997 that has helped to standardize testing. But this panel of markers has limitations resulting from the inclusion of dinucleotide markers, which are less sensitive and specific for detection of tumors with MMR deficiencies compared to other types of markers that are currently available. This study demonstrates that mononucleotides are the most sensitive and specific markers for detection of tumors with defects in MMR and identifies an optimal panel of markers for detection of MSI-H tumors. A set of 266 mono-, di-, tetra- and penta-nucleotide repeat microsatellite markers were used to screen for MSI in colorectal tumors. The best markers for detection of MSI-H tumors were selected for a MSI Multiplex System, which included five mononucleotide markers: BAT-25, BAT-26, NR-21, NR-24 and MONO-27. In addition, two pentanucleotide markers were added to identify sample mix-ups and/or contamination. We classified 153 colorectal tumors using the new MSI Multiplex System and compared the results to those obtained with a panel of 10 microsatellite markers combined with immunohistochemical (IHC) analysis. We observed 99% concordance between the two methods with nearly 100% accuracy in detection of MSI-H tumors. Approximately 5% of the MSI-H tumors had normal levels of four MMR proteins and as a result would have been misclassified based solely on IHC analysis, emphasizing the importance of performing MSI testing. The new MSI Multiplex System offers several distinct advantages over other methods of MSI testing in that it is both extremely sensitive and specific and amenable to high-throughput analysis. The MSI Multiplex System meets the new recommendations proposed at the recent 2002 NCI workshop on HNPCC and MSI testing and overcomes problems inherent to the original five-marker panel. The use of a single multiplex fluorescent MSI assay reduces the time and costs involved in MSI testing with increased reliability and accuracy and thus should facilitate widespread screening for microsatellite instability in tumors of patients with gastrointestinal cancers. PMID- 15528791 TI - MSI test to distinguish between HNPCC and other predisposing syndromes -- of value in tailored surveillance. PMID- 15528792 TI - Mutations associated with HNPCC predisposition -- Update of ICG-HNPCC/INSiGHT mutation database. AB - In 1994, the International Collaborative Group on Hereditary Nonpolyposis Colorectal Cancer (ICG-HNPCC) established an international database of mutations identified in families with Lynch (HNPCC) syndrome. The data are publicly available at http://www.nfdht.nl. The information stored in the database was systematically analyzed in 1997, and at that time, 126 different predisposing mutations were reported affecting the DNA mismatch repair genes MSH2 and MLH1 and occurring in 202 families. In 2003, the ICG-HNPCC and the Leeds Castle Polyposis Group (LCPG) merged into a new group, INSiGHT (International Society for Gastrointestinal Hereditary Tumors). The present update of the database of DNA mismatch repair gene mutations of INSiGHT includes 448 mutations that primarily involve MLH1 (50%), MSH2 (39%), and MSH6 (7%) and occur in 748 families from different parts of the world. PMID- 15528793 TI - Diagnostic application of hMLH1 methylation in hereditary non-polyposis colorectal cancer. AB - Colorectal cancer (CRC) due to mismatch repair (MMR) defect has distinct characteristics among unselected CRCs. These CRCs are biologically less aggressive and, thus, showing better prognosis but less sensitive to the 5FU based chemotherapy. CRCs with MMR defect derive from both hereditary and sporadic reasons. Germline inactivation of MMR genes (hMLH1, hMSH2, hMSH6, and hPMS2) underlies the hereditary CRC with MMR defect (Lynch syndrome) and epigenetic silencing of hMLH1 gene causes the sporadic CRC with MMR defect. Hereditary and sporadic CRC with MMR defect can be detectable by microsatellite instability (MSI) test or immunohistochemical analysis among general CRCs. Lynch syndrome can be diagnosed by the clinical criteria or by genetic test to detect pathogenic germline mutations in MMR genes. However, both clinical criteria and genetic test are inadequate for the diagnosis of Lynch syndrome. Since genetic test for the diagnosis of the Lynch syndrome is expensive and not always identify pathogenic germline mutations, effective and inexpensive screening program is desirable. Here we propose a possible application of methylation test combined with MSI or pathological analysis as an effective and a cost-saving new strategy for screening of Lynch syndrome. PMID- 15528795 TI - Ca[Ag2(SCN)4].2H2O. AB - Calcium tetrathiocyanatodiargentate(I) dihydrate, Ca[Ag(2)(SCN)(4)].2H(2)O, contains eight-membered Ag(4)S(4) rings bonded together through shared atoms to form layers parallel to (100). The thiocyanate groups link the layers to Ca-O chains running parallel to the c axis. The Ca atom is located on a twofold rotation axis parallel to b and is surrounded by four water molecules of crystallization and four thiocyanate N atoms in a distorted square antiprism. PMID- 15528794 TI - Neuroendocrine differentiation in sporadic CRC and hereditary nonpolyosis colorectal cancer. AB - Extent neuroendocrine differentiation can be encountered in many human neoplasm derived from different organs and systems using immunohistochemistry and ultrastructural techniques. The tumor cells' behaviors resemble those of neurons and neuroendocrine cells. The presence of neuroendocrine differentiation reputedly appears to be associated with a poorer prognosis than the adenocarcinoma counterparts in sporadic human neoplasm. In this review the neuroendocrine carcinoma and the adenocarcinoma with neuroendocrine differentiation of colon and rectum both in sporadic colorectal carcinoma and the hereditary nonpolyposis colorectal cancer, the relationship of neuroendocrine differentiation and some possible molecular pathways in tumorogenesis of colorectal cancer will be discussed. Possible treatment strategy will also be addressed. PMID- 15528796 TI - Pseudo-merohedrally twinned praseodymium hexacyanoferrate(III) tetrahydrate. AB - Crystals of the title compound, diaquahexa-mu-cyano-ferrate(III)praseodymium(III) dihydrate, Pr[Fe(CN)(6)].4H(2)O or [PrFe(CN)(6)(H(2)O)(2)].2H(2)O, are twinned with three components. The Pr atom is coordinated by eight atoms, viz. six N and two symmetry-related water O atoms. The Pr polyhedron (Pr has site symmetry m2m, Wyckoff position 4c) is linked to an FeC(6) octahedron (Fe located on a site with imposed 2/m symmetry, Wyckoff position 4b) through N atoms, forming an infinite array. The second (symmetry independent) water molecule lies on a mirror plane, is not included in coordination and is weakly hydrogen bonded to N atoms. PMID- 15528797 TI - beta-Tl2SO4. AB - The ambient-temperature form of dithallium sulfate, beta-Tl(2)SO(4), is similar to beta-K(2)SO(4) and is characterized by isolated sulfate tetrahedra and two different thallium sites with coordination numbers 9 and 11. All the atoms, except one O atom, lie on mirror planes. In spite of there being a high concentration of Tl(+) cations, the stereochemical activity of the 6s(2) pairs is low, similar to that of isotypic Tl(2)XO(4) compounds (X = Cr and Se). This behaviour is the consequence of both weak Tl-O bonds and strong X-O bonds, because in a Tl-O-X linkage the electronic cloud of the O(2-) anion is strongly distorted and displaced towards X, resulting in a low negative charge in the face of the Tl atom. Consequently, the Coulombic repulsions between the lone pair and the O(2-) anions are weak. All of the Tl(2)XO(4) compounds exhibit the same open packing of A(+) cations and [XO(4)](2-) anions as their isotypic alkali counterparts. PMID- 15528798 TI - Dicerium orthosilicate selenide and dicerium orthosilicate telluride, Ce2(SiO4)Q (Q = Se or Te). AB - The crystal structures of two new quaternary compounds, viz. dicerium orthosilicate selenide and dicerium orthosilicate telluride, Ce(2)(SiO(4))Q (Q = Se or Te), have been determined from single-crystal X-ray diffraction data. Each structure comprises infinite chains of SiO(4) tetrahedra separated by Ce and Q atoms. The site symmetries are Ce m and 2, Si 2 and Q m. The O atoms are in general positions. PMID- 15528799 TI - catena-Poly[[heptakis(dimethylformamide-kappaO)di-mu4-oxo-tetra-mu3-oxo-hexadeca mu2-oxo-tetraoxolanthanum(III)octamolybdenum(IV)]-mu-sodium(I)]. AB - The title compound, [NaLaMo(8)O(26)(C(3)H(7)NO)(7)](n), contains infinite chains of [Mo(8)O(26)](4-) units supporting dimethylformamide-coordinated La(III) cations and linked by Na(+) cations. The lanthanum center adopts a nine coordinate geometry and the Na atom is sandwiched between two beta-[Mo(8)O(26)](4 ) units. PMID- 15528800 TI - Potassium hydrogen diphthalate dihydrate: a new structure and correction to the literature. AB - The title compound, K(+).[(C(8)H(5)O(4))(2)H](-).2H(2)O or K(+).C(16)H(11)O(8)( ).2H(2)O, was prepared by slow evaporation of an aqueous solution of potassium hydrogen phthalate. The molecular complex consists of a potassium cation coordinated to a proton-bound hydrogen phthalate dimer and two water molecules. The potassium cation resides on a twofold axis in a distorted square-antiprism coordination geometry. The compound is isomorphous with the ammonium analogue, previously misidentified. As potassium hydrogen phthalate is frequently used in the manufacture of buffers, organic carbon standards, acidimetric standards and various other products, the crystallization of a compound with a different stoichiometery from a solution containing the acidimetric standard has important practical implications. PMID- 15528801 TI - catena-Poly[[bis(pentane-2,4-dionato-kappa2O,O')cobalt(II)]-mu-4,4' methylenedianiline-kappa2N:N']. AB - In the title compound, [Co(C(5)H(7)O(2))(2)(C(13)H(14)N(2))](n) or [Co(acac)(2)(dadpm)](n), where acac is acetylacetonate and dadpm is 4,4' methylenedianiline, the Co atom is on a centre of symmetry and is octahedrally coordinated by four O atoms from two acac anions and by two N atoms from two dadpm ligands. Each dadpm ligand, which has a twofold axis passing through its methylene C atom, bridges two Co atoms to form a spiral polymeric chain. Neighbouring chains connect via hydrogen bonds to form a two-dimensional network. PMID- 15528802 TI - A one-dimensional carboxylate-bridged helical copper(II) complex containing (quinolin-8-yloxy)acetate. AB - The title compound, catena-poly[[bromocopper(II)]-mu-(quinolin-8-yloxy)acetato kappa(4)N,O,O':O''], [CuBr(C(11)H(8)NO(3))](n), is a novel carboxylate-bridged one-dimensional helical copper(II) polymer. The metal ion exhibits an approximately square-pyramidal CuBrNO(3) coordination environment, with the three donor atoms of the ligand and the bromide ion occupying the basal positions, and an O atom belonging to the carboxylate group of an adjacent molecule in the apical site. Carboxylate groups are mutually cis oriented, and each anti-anti carboxylate group bridges two copper(II) ions via one apical and one basal position [Cu...Cu = 5.677 (1) A], resulting in the formation of a helical chain along the crystallographic b axis. PMID- 15528803 TI - A novel two-dimensional cobalt(II) coordination polymer with 1,4-bis(1,2,4 triazol-1-ylmethyl)benzene. AB - In the crystal structure of the title complex, poly[[diazidocobalt(II)]-di-mu-1,4 bis(1,2,4-triazol-1-ylmethyl)benzene-kappa(4)N(4):N(4')], [Co(N(3))(2)(bbtz)(2)](n), where bbtz is 1,4-bis(1,2,4-triazol-1-ylmethyl)benzene (C(12)H(12)N(6)), the Co(II) atom, which lies on an inversion centre, is six coordinated by four N atoms from four bbtz ligands and by two N atoms from two azide ligands, in a distorted octahedral coordination environment. The Co(II) atoms are bridged by four bbtz ligands to form a two-dimensional [4,4]-network. PMID- 15528804 TI - mer-Diaquabis(1H-imidazole-kappaN3)(orotato-kappa2N3,O4nickel(II). AB - The title mononuclear complex, [Ni(C(5)H(2)N(2)O(4))(C(3)H(4)N(2))(2)(H(2)O)(2)] or [Ni(HOr)(im)(2)(H(2)O)(2)] (im is imidazole and H(3)Or is orotic acid, or 2,6 dioxo-1,2,3,6-tetrahydropyrimidine-4-carboxylic acid), has been synthesized and the crystal structure determination is reported. The Ni(II) ion in the complex has a distorted octahedral coordination geometry comprised of one deprotonated pyrimidine N atom and the adjacent carboxylate O atom of the orotate ligand, two tertiary imidazole N atoms and two aqua ligands. An extensive three-dimensional network of OW-H...O and N-H...O hydrogen bonds, and pi-pi and pi-ring interactions are responsible for crystal stabilization. PMID- 15528805 TI - catena-Poly[[di-mu-benzoato-kappa4O:O'-disilver(I)]-mu-N,N'-bis(2 fluorobenzylidene)butane-1,4-diamine-kappa2N:N']. AB - The title complex, [Ag(2)(C(7)H(5)O(2))(2)(C(18)H(18)F(2)N(2))](n), is a dinuclear silver(I) compound with one inversion centre between pairs of Ag atoms and another at the mid-point of the central C-C bond in the butane-1,4-diamine moiety. Each of the smallest repeat units consists of two silver(I) cations, two benzoate anions and one N,N'-bis(2-fluorobenzylidene)butane-1,4-diamine Schiff base ligand. Each Ag(I) ion is three-coordinated in a trigonal configuration by two O atoms from two benzoate anions and one N atom from a Schiff base ligand. The di-mu-benzoato-disilver(I) moieties are linked by the bridging Schiff base ligand, giving zigzag polymeric chains with an [-Ag...Ag-N-C-C-C-C-N-](n) backbone running along the b axis. PMID- 15528806 TI - Poly[[mu2-aqua-bis[(1,10-phenanthroline)nickel(II)]]-di-mu2,mu4-5-nitro-1,3 benzenedicarboxylato]. AB - The reaction of Ni(CH(3)COO)(2).4H(2)O, 5-nitro-1,3-benzenedicarboxylic acid (H(2)nmbdc), 1,10-phenanthroline and water under hydrothermal conditions yields the first reported two-dimensional nickel coordination polymer with water- and carboxylate-bridged dimeric units, viz. [Ni(2)(C(8)H(3)NO(6))(2)(C(12)H(8)N(2))(2)(H(2)O)](n). The coordination polyhedron of the Ni(II) ion in the title structure is an octahedron defined by an N(2)O(4) donor set. The water molecule is positioned on a mirror plane and the 5-nitro-1,3-benzenedicarboxylate group is located on a twofold axis. Two types of nmbdc(2-) coordination mode are observed: one is a bis-monodentate mode, mu(2) nmbdc(2-), and the other is a bis-bridging mode, mu(4)-nmbdc(2-). The dimeric unit in the title compound is similar to the structural moiety in urease. In the two-dimensional framework in the title compound, strong stacking interactions between benzene rings (mu(2)-nmbdc(2-) and mu(4)-nmbdc(2-)) and 1,10 phenanthroline ligands are observed. PMID- 15528807 TI - Diazabicyclo[2.2.2]octane-1,4-diium dichromate. AB - The title compound, (C(6)H(14)N(2))[Cr(2)O(7)], consists of a diazabicyclo[2.2.2]octane-1,4-diium cation and a discrete dichromate anion, which are linked in the crystal by N-H...O hydrogen bonds. The cation is ordered and distorted, owing to the confinement and twist of the hydrogen bonds involved. Two CrO(4) tetrahedra are joined through a shared O atom to form the dichromate anion. Chiral supramolecular chains of the title compound are built up via N H...O hydrogen bonds, and C-H...O interactions play subordinate roles in forming the structure. PMID- 15528808 TI - catena-Poly[[chloro(1,10-phenanthroline-kappa2N,N')copper(II)]-mu-imidazolato kappa2N:N']. AB - In the polymeric title compound, [Cu(im)Cl(phen)](n), where im is the imidazolate anion (C(3)H(3)N(2)) and phen is 1,10-phenanthroline (C(12)H(8)N(2)), each Cu(II) ion is five-coordinated by four basal N atoms (two from two different im anions and two from one phen ligand) and one axial Cl atom, in a distorted square pyramidal coordination geometry. Moreover, each im anion bridges two identical [CuCl(phen)](+) cations through its two N atoms, resulting in a one-dimensional zigzag chain along the crystallographic a axis. In addition, pairs of adjacent chains are staggered by pi-pi interactions, generating a two-dimensional layer, and neighbouring layers are further linked by two different kinds of C-H...Cl interactions, producing a three-dimensional network. PMID- 15528809 TI - [4-Bromo-2-[2-(5-bromo-2-oxidobenzylideneaminomethyl)phenyliminomethyl]phenolato kappa4O,N,N',O']nickel(II). AB - In the title complex, [Ni(C(21)H(14)Br(2)N(2)O(2))], the Ni(II) atom is coordinated by the two imine N and two phenolate O atoms of the Schiff base ligand in a tetrahedrally distorted square-planar geometry. The Ni-N and Ni-O distances are within the ranges expected for Ni-Schiff base derivatives. Intermolecular C-H...O hydrogen bonds link the molecules into centrosymmetric dimers, forming R(2)(2)(12) (A) and R(2)(2)(10) (B) rings. These dimers combine to form a supramolecular ABAB... aggregate which propagates along the [100] direction. PMID- 15528810 TI - Polymeric mu-cyano-dicyanonickelate(II)-mu-cyano-trans-bis[N-(2 hydroxyethyl)ethylenediamine]cadmium(II). AB - The title compound, catena-poly[[mu-cyano-1:2kappa(2)C:N-dicyano-1kappa(2)C-trans bis[N-(2-hydroxyethyl)ethane-1,2-diamine-2kappa(2)N,N']cadmium(II)nickel(II)]-mu cyano-1:2'kappa(2)C:N], [CdNi(CN)(4)(C(4)H(12)N(2)O)(2)], consists of alternating square-planar Ni(CN)(4) fragments, formally dianionic, and Cd(hydet-en)(2) moieties [hydet-en is N-(2-hydroxyethyl)ethylenediamine], with the two bridging cyanide ligands in a mutually trans disposition at the Ni atom and cis at the Cd atom. The resulting one-dimensional zigzag chain structure has the Ni atom on an inversion center, while the distorted octahedron centered on the Cd atom lies on a twofold axis. The polymer chains are connected into undulating sheets by weak interchain N-H...N, N-H...O and O-H...N hydrogen bonds, which are also present between successive sheets. PMID- 15528811 TI - [2,6-Bis[(dimethylamino)methyl]pyridine-kappa3N](2,2-methylenediphenolato kappa2O,O')dioxouranium(VI) acetone solvate. AB - In the title compound, [UO(2)(C(13)H(10)O(2))(C(11)H(19)N(3))].C(3)H(6)O, the U atom is in a pentagonal-bipyramidal environment, with the three N atoms of the 2,6-bis[(dimethylamino)methyl]pyridine ligand and the two O atoms of the dianionic 2,2'-methylenediphenolate ligand in the equatorial plane. The geometry is compared with that of previously reported 1:2 uranyl-diphenoxide complexes. PMID- 15528812 TI - Revisited crystal symmetry of the high-spin form of the iron(II) spin-crossover complex dicyano[2,13-dimethyl-6,9-dioxa-3,12,18-triazabicyclo[12.3.1]octadeca 1(18),2,12,14,16-pentaene]iron(II) monohydrate. AB - The title iron(II) complex, [Fe(CN)(2)(C(15)H(23)N(3)O(2))].H(2)O, is of interest to the spin-crossover community because of its unusual temperature-dependent magnetic behaviour as well as its relatively high relaxation temperature for the light-induced spin-crossover phenomenon. Structural modifications are strongly suspected to cause the unusual thermal spin-crossover features. Recently, the high-spin crystal structure has been reported but with an inadequate space group. In the present paper, the crystal structure is corrected by a new investigation, and some consequences for the structure-property relationships of this complex are discussed. The Fe(II) ion is seven-coordinate and lies on a twofold axis. PMID- 15528813 TI - catena-Poly[[diaquacadmium(II)]-mu-5-carboxyimidazole-4-carboxylato kappa4N1,O5:O4,N3]. AB - A new cadmium coordination polymer, [Cd(C(5)H(2)N(2)O(4))(H(2)O)(2)](n), possesses a one-dimensional zigzag chain structure built from Cd(II) centers bridged sequentially by pairs of O and N atoms of the 5-carboxyimidazole-4 carboxylate ligand. The Cd(II) center is in a distorted octahedral geometry, being coordinated by two O atoms from two coordinated water molecules [Cd-O = 2.322 (7) and 2.364 (7) A], and by two N atoms [Cd-N = 2.222 (6) and 2.232 (6) A] and two carboxyl O atoms [Cd-O = 2.383 (6) and 2.414 (6) A] from two 5 carboxyimidazole-4-carboxylate ligands. PMID- 15528814 TI - Poly[[tetraaqua(mu7-hydrogen dichloromethylenebisphosphonato)(mu5-hydrogen dichloromethylenebisphosphonato)tribarium(II)] monohydrate]. AB - The title compound, [[Ba(3)(CHCl(2)O(6)P(2))(2)(H(2)O)(4)].H(2)O](n) or [[Ba(3)(Cl(2)CP(2)O(6)H)(H(2)O)(4)].H(2)O](n), is two-dimensional. The asymmetric unit contains three independent Ba(2+) atoms, two chelating and bridging Cl(2)CP(2)O(6)H(3-) ligands and four aqua ligands, connected in layers parallel to the (100) plane. There are pores between the layers in the direction of the b axis filled with lattice water molecules. PMID- 15528815 TI - Bis(2,2'-bipyridine-kappa2N,N')(oxalato-kappa2O,O')cobalt(II) pentahydrate. AB - The synthesis and crystal structure of the mononuclear title compound, [Co(C(2)O(4))(C(10)H(8)N(2))(2)].5H(2)O, is reported. The Co atom is six coordinated by two O atoms of a bidentate oxalate group and by four N atoms of two bipyridine ligands. The neutral [Co(C(2)O(4))(C(10)H(8)N(2))(2)] entities are connected by pi-pi stacking interactions of the aromatic systems into a two dimensional layer, interconnected through a ladder-like hydrogen-bonding pattern of solvate water molecules. PMID- 15528816 TI - Self-assembly of a mixed-ligand silver-based coordination polymer. AB - The novel title silver(I) coordination polymer, catena poly[[acetonitrilesilver(I)]-di-mu-4-[N-(diphenylphosphino)aminomethyl]pyridine kappa(2)N(1):P;kappa(2)P:N(1)-[acetonitrilesilver(I)]-mu(3)-4-[N,N bis(diphenylphosphino)aminomethyl]pyridine-kappa(3)N(1):P:P' bis[acetonitrilesilver(I)(Ag-Ag)]-mu(3)-4-[N,N bis(diphenylphosphino)aminomethyl]pyridine-kappa(3)P:P':N(1)] tetrakis(tetrafluoroborate) acetonitrile trisolvate], [[Ag(4)(C(2)H(3)N)(4)(C(18)H(17)N(2)P)(2)(C(30)H(26)N(2)P(2))(2)](BF(4))(4).3C(2) (3)N](n), is formed by the self-assembly of the Ph(2)P(4-NHCH(2)C(5)H(4)N) and (Ph(2)P)(2)(4-NCH(2)C(5)H(4)N) ligands with silver tetrafluoroborate. The polymer consists of alternating rings (which lie about inversion centers) bridged by the pyridyl rings of the bis-phosphine-substituted ligands, with anions hydrogen bonded the length of the chain. Two distinctly different metal coordination environments exist in the polymer, viz. distorted tetrahedral and trigonal geometries. PMID- 15528817 TI - (4R,4aR,6S,7S,7aS)-6-Hydroxy-7-hydroxymethyl-4-methylperhydrocyclopenta[c]pyran-1 one chloroform solvate from Valeriana laxiflora. AB - The structure of an iridolactone isolated from Valeriana laxiflora was established as (4R,4aR,6S,7S,7aS)-6-hydroxy-7-hydroxymethyl-4 methylperhydrocyclopenta[c]pyran-1-one chloroform solvate, C(10)H(16)O(4).CHCl(3). The two rings are cis-fused. The delta-lactone ring adopts a slightly twisted half-chair conformation with approximate planarity of the lactone group and the cyclopentane ring adopts an envelope conformation. The hydroxy group, the hydroxymethyl group and the methyl group all have beta orientations. The absolute configuration was determined using anomalous dispersion data enhanced by the adventitious inclusion of a chloroform solvent molecule. Hydrogen bonding, crystal packing and ring conformations are discussed in detail. PMID- 15528818 TI - 5,5'-Di-tert-butyl-2,2'-dihydroxy-3,3'-methanediyldibenzaldehyde and its allyl protected dialcohol and dialdehyde precursors. AB - 5,5'-Di-tert-butyl-2,2'-dihydroxy-3,3'-methanediyldibenzaldehyde, C(23)H(28)O(4), (IV), has been structurally characterized in two polymorphic forms. The tetragonal form, (in I4(1)/a) has been reported previously but is redetermined and reinterpreted here, while the monoclinic form, (in C2/c) is reported for the first time. In both polymorphs, the molecule lies on a crystallographic twofold axis. Two precursors in the synthesis of (IV), namely 2,2'-bis(allyloxy)-5,5'-di tert-butyl-3,3'-methanediyldibenzenemethanol (C(29)H(40)O(4)) and 2,2' bis(allyloxy)-5,5'-di-tert-butyl-3,3'-methanediyldibenzaldehyde (C(29)H(36)O(4)) have also been characterized. PMID- 15528819 TI - Polymorphism and solvolysis of 2-cyano-3-[4-(N,N-diethylamino)phenyl]prop-2 enethioamide. AB - Two new polymorph forms, (Ia) and (Ib), of the title compound, C(14)H(17)N(3)S, and its solvate with acetonitrile, C(14)H(17)N(3)S.0.25C(2)H(3)N, (Ic), have been investigated. Crystals of the two polymorphs were grown from different solvents, viz. ethanol and N,N-dimethylformamide, respectively. The polymorphs have different orientations of the thioamide group relative to the CN substituent, with s-cis and s-trans geometry of the C=C-C=S diene fragment, respectively. Compound (Ic) contains two independent molecules, A and B, with s-cis geometry, and the solvate molecule lies on a twofold axis. The core of each molecule is slightly non-planar; the dihedral angles between the conjugated C=C-CN linkage and the phenyl ring, and between this linkage and the thioamide group are 13.4 (2) and 12.0 (2) degrees in (Ia), 14.0 (2) and 18.2 (2) degrees in (Ib), 2.3 (3) and 12.7 (4) degrees in molecule A of (Ic), and 23.2 (3) and 8.1 (4) degrees in molecule B of (Ic). As a result of strong conjugation between donor and acceptor parts, the substituted phenyl rings have noticeable quinoid character. In (Ib), there exists a very strong intramolecular steric interaction (H...H = 1.95 A) between the bridging and thioamide parts of the molecule, which makes such a form less stable. In the crystal structure of (Ia), intermolecular N-H...N and N-H...S hydrogen bonds link molecules into infinite tapes along the [1-10] direction. In (Ib), such intermolecular hydrogen bonds link molecules into infinite (101) planes. In (Ic), intermolecular N-H...N hydrogen bonds link molecules A and B into dimers, which are connected via N-H...S hydrogen bonds and form infinite chains along the c direction. PMID- 15528820 TI - (2S)-2-[1-(2,4-Difluoro-5-iodophenyl)-2-deoxy-beta-D-ribofuranos-5-yloxy]-8 methyl-4H-1,3,2-benzodioxaphosphole 2-oxide. AB - The title compound, C(19)H(18)F(2)IO(6)P, prepared as a potential antiviral and anticancer agent from 3-methylsalicylchlorophosphane and 1-(2,4-difluoro-5 iodophenyl)-2-deoxy-beta-D-ribofuranose, is one of a 1:1 mixture of two diastereomers. The diastereomers differ in their configuration, S or R, at the asymmetric phosphorus center. X-Ray crystallographic analysis of the title compound has determined the absolute configuration at the asymmetric P center to be S. PMID- 15528821 TI - Catharanthinol and dihydrocatharanthinol: two Iboga-class alkaloids. AB - The title compounds are indole alkaloids of the Iboga class. In both compounds, viz. catharanthinol methanol solvate, C(20)H(24)N(2)O.CH(4)O, (I), and dihydrocatharanthinol monohydrate, C(20)H(26)N(2)O.H(2)O, (II), a nitrogen containing seven-membered ring is fused to the indole system and shares two sides with a tricyclic isoquinuclidine group. The main difference between (I) and (II) is the presence of a C=C bond in the isoquinuclidine ring in (I). The presence of amine and hydroxy groups in these molecules and of methanol [in (I)] or water [in (II)] solvent molecules results in intra- and/or intermolecular hydrogen bonding. PMID- 15528822 TI - N-(3-Methyl-4-oxo-3,4-dihydropteridin-2-yl)glycine: hydrogen-bonded sheets of R44(22) and R44(30) rings. AB - Molecules of the title compound, C(9)H(9)N(5)O(3), are linked into sheets by a combination of one O-H...N hydrogen bond and one N-H...O hydrogen bond. PMID- 15528823 TI - Mikanolide from Jamaican Mikania micrantha. AB - Mikanolide [systematic names: 1,10:2,3-diepoxy-6,8-dihydroxy-11-vinylgermacr-4 ene 12,14-di-gamma-lactone and 7,10a-dimethyl-1a,1b,2a,6a,7,9a,10,10a-octahydro 4H-6,3-methenofuro[3,2-c]bisoxireno[f,h]oxacycloundecin-4,8(6H)-dione], C(15)H(14)O(6), derived from a variety of Mikania micrantha growing in Portland, Jamaica, contains a methylcyclodecane ring fused to an unsaturated planar alpha,gamma-lactone, an envelope-type near-planar vinyl-beta,gamma-lactone and two epoxide moieties. The crystal packing shows stacks of mikanolide molecules interlocked via a network of non-classical C-H...O hydrogen bonds between the lactone units. PMID- 15528824 TI - Racemic 3,5-dichloro-2-[[(1-phenylethyl)imino]methyl]phenol. AB - The title compound, (RS)-3,5-dichloro-2-[[(1-phenylethyl)imino]methyl]phenol, C(15)H(13)Cl(2)NO, was synthesized from racemic 1-phenylethylamine and 3,5 dichlorosalicylaldehyde. The pi-conjugate system around the imine group is essentially planar in the phenol-imine tautomer. Intramolecular O...N hydrogen bond and intermolecular C-H...pi interactions are present in the crystal structure. PMID- 15528825 TI - 1,3-Bis[2-(2-hydroxybenzylideneamino)phenoxy]propane. AB - The title compound [systematic name: 2,2'-[1,3-propanediyldioxydi-o phenylenebis(nitrilomethylidyne)]diphenol], C(29)H(26)N(2)O(4), exists as the phenol-imine form in the crystal, and there are strong intramolecular O-H...N hydrogen bonds, with O...N distances of 2.545 (2) and 2.579 (2) A. The C=N imine bond distances are in the range 1.276 (2)-1.279 (2) A and the C=N-C bond angles are in the range 123.05 (16)-124.64 (17) degrees . The configurations about the C=N bonds are anti (1E). PMID- 15528826 TI - 3,5-Bis(4-methoxybenzylidene)-1-methyl-4-piperidone and 3,5-bis(4 methoxybenzylidene)-1-methyl-4-oxopiperidinium chloride: potential biophotonic materials. AB - In the title compound 3,5-bis(4-methoxybenzylidene)-1-methyl-4-piperidone, C(22)H(23)NO(3), (I), the central heterocyclic ring adopts a flattened boat conformation, while in the related salt 3,5-bis(4-methoxybenzylidene)-1-methyl-4 oxopiperidinium chloride, C(22)H(24)NO(3)(+).Cl(-), (II), the ring exhibits a 'sofa' conformation in which the N atom deviates from the planar fragment. The pendant benzene rings are twisted from the heterocyclic ring planes in both molecules in the same direction, the range of dihedral angles between the ring planes being 24.5 (2)-32.7 (2) degrees . The dominant packing motif in (I) involves centrosymmetric dimers bound by weak intermolecular C-H...O hydrogen bonds. In (II), cations and anions are linked by strong N-H...Cl hydrogen bonds, while weak C-H...O and C-H...Cl hydrogen bonds link the cations and anions into a three-dimensional framework. PMID- 15528827 TI - L-Phenylalanyl-L-isoleucine 0.88-hydrate. AB - The asymmetric unit in the crystal structure of the title compound, C(15)H(22)N(2)O(3).0.88H(2)O, contains two peptide molecules with completely different conformations. The structure is divided into hydrophobic and hydrophilic layers, with channels of water molecules at the layer interface. PMID- 15528829 TI - 2-Acetamido-4-p-tolyl-1,3-thiazole and 2-amino-4-p-tolyl-1,3-thiazolium chloride dihydrate. AB - The structures of 2-acetamido-4-tolyl-1,3-thiazole, C(12)H(12)N(2)OS, (I), and 2 amino-4-tolyl-1,3-thiazolium chloride dihydrate, C(10)H(11)N(2)S(+).Cl(-).2H(2)O, (II), reveal that both molecules are essentially planar, with the respective dihedral angles between the benzene and thiazole rings being 2.9 (1) and 10.39 (7) degrees . Compound (I) associates via a single N-H...O interaction to form a flat alternate-facing hydrogen-bonded chain [graph-set C(2)(2)(4)]. Compound (II) packs with the hydrogen-bonding associations of the Cl atoms and the water molecules creating a convoluted hydrogen-bonded ribbon made up of five-membered donor-acceptor rings, involving three water O atoms (with associated H atoms) and two Cl atoms. The thiazolium rings form stacked columns, aligned in the same direction as the hydrogen-bonded ribbons, of alternate-facing molecules that are also involved in the hydrogen-bonding network, linking to the Cl atoms and one of the water molecules. Subsequently, each Cl atom is the hydrogen-bond acceptor for five separate O/N-H associations. PMID- 15528828 TI - 3-Benzyloxy-16-[(N-methyl-N-phenylamino)methylidene]estra-1,3,5(10)-trien-17-one. AB - In the title compound, C(33)H(35)NO(2), the five-membered ring adopts a half chair conformation. The N-methyl-N-phenyl-substituted keto-enamine moiety shows a comparatively long Csp(2)-N bond. PMID- 15528830 TI - 3-Phenyl-4H,6H-1,2,4-oxadiazol-5-one. AB - Molecules of the title compound, C(9)H(8)N(2)O(2), are linked into complex sheets by a combination of N-H...O, C-H...N and C-H...O hydrogen bonds. PMID- 15528831 TI - Trityl losartan. AB - The title compound (systematic name: [2-butyl-4-chloro-1-[2'-(2-trityl-2H tetrazol-5-yl)biphenyl-4-ylmethyl]-1H-imidazol-5-yl]methanol), C(41)H(37)ClN(6)O, crystallizes in the centrosymmetric space group P-1 with two independent molecules in the asymmetric unit. These molecules differ significantly only in the relative orientations of the rings in the biphenylyltetrazole moieties. One of the molecules shows disorder for three C atoms in the n-butyl group. Hydrogen bonds link the molecules in an infinite chain along the a axis. PMID- 15528832 TI - Two new polymorphs of diphenyl(4-pyridyl)methyl methacrylate. AB - The title compound (D4PyMA), C(22)H(19)NO(2), exhibits polymorphism after crystallization by slow evaporation from a binary mixture of chloroform and hexane. Long needle-like crystals have an orthorhombic structure (space group Fdd2), with one molecule in the asymmetric unit, while small tablet-like crystals exhibit a monoclinic crystal structure (space group P2(1)/n), in which two independent but chemically identical molecules comprise the asymmetric unit. The bond lengths and angles are normal, while the torsion angles around the -C-O- bond linking the diphenyl(4-pyridyl)methyl and methacrylate groups show the flexibility of the molecule by way of packing effects. The two polymorphs both contain weak C-H...pi and C-H...O/N contacts but have different conformations. PMID- 15528833 TI - 6-(4-Bromophenyl)-6,7-dihydro-1,3-dioxolo[4,5-g]quinolin-8(5H)-one: bilayers built from N-H...O, C-H...O and C-H...pi(arene) hydrogen bonds. AB - Molecules of the title compound, C(16)H(12)BrNO(3), exhibit a polarized molecular electronic structure. A combination of one N-H...O hydrogen bond and one C-H...O hydrogen bond links the molecules into sheets, and pairs of sheets are linked into bilayers by a single C-H...pi(arene) hydrogen bond. PMID- 15528834 TI - (4R)-3-Allenyl-4-(diphenylmethyl)oxazolidin-2-one, an unsubstituted allenamide. AB - The first X-ray structure of an unsubstituted allenamide, C(19)H(17)NO(2), is reported. The solid-state phase supports the notion that a key minimum conformation of allenamides can be invoked to rationalize the observed stereochemical outcomes in many of our methodological studies employing allenamides. This minimum conformation involves two important factors, i.e. having approximate coplanarity between the planes of the oxazolidinone ring and the internal olefin, and having the allene moiety facing away from the carbamate carbonyl group. The C-N-C=C torsion angle that quantifies this approximate coplanarity between the plane of the oxazolidinone ring and that of the internal olefin, as determined from this crystallographic study, is -19.1 (2) degrees . A minimized structural calculation, which determined this angle to be -16.1 degrees , is in close agreement. Additional structural features include a probable pi-pi interaction between the allene moiety and a benzene ring, and non-classical hydrogen bonding in the form of weak C-H...O interactions that are responsible for the formation of two-dimensional networks. PMID- 15528835 TI - Candibirin A, a furanocoumarin dimer isolated from Heracleum candicans WALL. AB - Candibirin A [systematic name: 9,9'-(1,4-dioxane-2,5-diyldimethylenedioxy)di(7H furo[3,2-g]chromen-7-one)], a new furanocoumarin dimer, was isolated from Heracleum candicans WALL. (1)H NMR and MS spectra had indicated that the title compound was a dimer of heraclenin or heraclenol, but the linkage structure and its chirality were undetermined. The dioxane linkage, having the R,R configuration, has now been elucidated from dimethyl sulfoxide-solvated crystals, C(32)H(28)O(10).2C(2)H(6)OS. Candibirin A is thus a dimerization product from heraclenin formed by reaction at the epoxy group. Dimethylformamide-solvated crystals, C(32)H(28)O(10).C(3)H(7)NO, adopt a different conformation, with a folded structure that differs from the extended structure in the dimethyl sulfoxide solvate. However, the puckering of the dioxane linker unit is very similar in the two conformers. This result shows that the rotation of the ether bonds, in the linker between the furanocoumarin and dioxane moieties, causes the conformational flexibility of (I). PMID- 15528836 TI - 8,19-Dimethyl-8,19-dihydro-8,19-ethanoanthra[2'',3'':2,3;6'',7'':2',3']di-1,4 dioxino[5,6-b;5',6'-b']diquinoxaline. AB - The title compound, C(34)H(22)N(4)O(4), results from the reaction of 2,3,6,7 tetrahydroxy-9,10-dimethyl-9,10-dihydro-9,10-ethanoanthracene with 2,3 dichloroquinoxaline. The molecule, which contains a binary crystallographic symmetry axis, comprises two planar 'wings' around a central bicyclic unit. The non-ideal geometry of the latter evidences some strain, as in previous compounds with the same central core. Each molecule is involved in pi-pi interactions with four of its neighbours, oriented upside-down, which results in the formation of sheets of tightly packed molecules. PMID- 15528837 TI - The 2:1 adducts of (benzoylmethylene)triphenylphosphorane with fumaric and terephthalic acids. AB - Co-crystals of the ylide (benzoylmethylene)triphenylphosphorane (BPPY) with either fumaric acid, viz. (benzoylmethylene)triphenylphosphorane-fumaric acid (2/1), C(26)H(21)OP.0.5C(4)H(4)O(4), or terephthalic acid, viz. (benzoylmethylene)triphenylphosphorane-terephthalic acid (2/1), C(26)H(21)OP.0.5C(8)H(6)O(4), have a stoichiometric ratio of 2:1 between the ylide and the corresponding dicarboxylic acid. In both adducts, the acid component lies across a centre of inversion. In neither case is the ylide protonated by the organic acid; instead the H atoms of the non-ionized dicarboxylic acid molecules participate in the formation of strong O-H...O hydrogen bonds with the benzoyl O atom of the ylide species. These structures are the first reported examples of co-crystals containing non-protonated BPPY. PMID- 15528838 TI - Turnover of actin in Chlamydomonas flagella detected by fluorescence recovery after photobleaching (FRAP). AB - Recent indirect observations have suggested that various axonemal proteins in cilia and flagella of live cells undergo turnover independently of shortening or elongation of the axoneme. To gain direct evidence, here we examined using a FRAP (fluorescence recovery after photobleaching) technique whether actin, a subunit of inner arm dynein, is being turned over in Chlamydomonas flagella. Fluorescently labeled rabbit actin was introduced by electroporation into the cells of ida5oda1, a double mutant between oda1 lacking outer arm dynein and ida5 lacking several species of inner arm dyneins due to the absence of a conventional type actin. In actin-loaded cells, flagella became motile and fluorescent due to incorporation of inner-arm dyneins containing the labeled actin. Cells were sandwiched between an agar layer and a coverslip so as to restrict flagellar movement. After a small portion of a flagellum was photobleached, the fluorescence intensity in the bleached area was monitored with a sensitive video camera. The fluorescence intensity in the photobleached region was found to recover 10-40% of the original level over several tens of minutes without changing its position. The time course and extent of the recovery varied greatly from one cell to another, suggesting that the turnover depends on cellular conditions. Western blot analysis indicated that 70-80% of flagellar actin was associated with the axoneme. Hence this experiment provides direct evidence that an axonemal component undergoes dynamic exchange in stationary flagella. PMID- 15528839 TI - Human amniotic epithelial cells possess hepatocyte-like characteristics and functions. AB - Hepatocyte transplantation is expected to become a novel method for treatment of liver disease. However, many questions remain regarding this approach, especially concerning donor cells. To evaluate whether human amniotic epithelial cells can be used as a cell source for hepatocyte transplantation, hepatic gene expression and functions of human amniotic epithelial cells were analyzed. Reverse transcription-polymerase chain reaction analysis demonstrated that human amniotic epithelial cells expressed albumin, alpha(1)-antitrypsin, and other hepatocyte related genes. Cultivated human amniotic epithelial cells demonstrated albumin production, glycogen storage, and albumin secretion consistent with the hepatocyte gene expression profile. In organ culture, the amnion secreted 30-fold larger amounts of albumin than human amniotic epithelial cells in monolayer culture. Moreover, in organ culture the amnion also secreted alpha(1) antitrypsin. Following transplantation into mice, the amnion survived and secreted albumin. These observations suggest that transplantation of human amniotic epithelial cells and/or amnion could be novel therapeutic strategy for treatment of hepatic diseases, including alpha(1)-antitrypsin deficiency. PMID- 15528840 TI - Diphenidol has no actual broad antiemetic activity in dogs and ferrets. AB - Previous studies showed that diphenidol was effective on emetogens-induced pica, eating of non-nutritive substances, in rats, a model analogous to emesis in other species. We evaluated the actual antiemetic activity of diphenidol against four emetic stimuli in the dog and ferret, animals that possess an emetic reflex. In dogs, emetic responses to apomorphine were significantly prevented by diphenidol (3.2 mg/kg, i.v.), whereas diphenidol (3.2 mg/kg, i.v. x 2) showed a weak inhibition to the vomiting evoked by cisplatin. In ferrets, diphenidol (10 mg/kg, i.p.) exhibited a weak antiemetic activity on the emesis induced by copper sulfate and had no activity on emesis by loperamide. On the other hand, CP 122,721, a NK1-receptor antagonist, significantly reduced the emetic episodes to all four stimuli. These results suggest that the prediction of antiemetic activity of compounds in animals lacking an emetic reflex does not always correspond with actual antiemetic activity. PMID- 15528841 TI - Inhibitory effect of antituberculosis drugs on human cytochrome P450-mediated activities. AB - The potential for drug-drug interactions mediated by the inhibition of cytochrome P-450 (CYP) were concerned during antituberculosis therapy. However, the information regarding human CYP inhibition by antituberculosis drugs is limited to isoniazid. In the current study, we examined the inhibitory effects of pyrazinamide and ethionamide, both of which are chemically related to isoniazid, on the CYP-mediated activities in human liver microsomes and compared them to that of isoniazid. No remarkable effects on any CYP activities were observed by pyrazinamide and ethionamide. In contrast, in addition to the reported inhibitory effect of isoniazid on CYP1A2, CYP2A6, CYP2C19, and CYP3A activities, our results newly showed its effect on CYP2C9 and CYP2E1 activities. Isoniazid showed potent direct inhibitory effect on S-warfarin 7-hydroxylation, while a preincubation step in the presence of NADPH was needed to inhibit chlorzoxazone 6 hydroxylation. Furthermore, irreversible inhibition of CYP2C19 activity by isoniazid was also observed in the dilution study. These results suggested that pyrazinamide and ethionamide did not seem to cause drug interactions mediated by the inhibition of CYP. In contrast, isoniazid might contribute to the severe drug interactions by a different inhibitory mechanism depending on each of the CYP isozymes, in addition to the reported observations. PMID- 15528842 TI - Role of K+ channels in M2 muscarinic receptor-mediated inhibition of noradrenaline release from the rat stomach. AB - Previously we reported the cholinergic M2 muscarinic receptor-mediated inhibition of noradrenaline release from the rat stomach (K. Yokotani, Y. Osumi. J Pharmacol Exp Ther. 1993;264:54-60). In the present study, we investigated the role of K+ channels in oxotremorine (a muscarinic receptor agonist)-induced inhibition of noradrenaline release using isolated, vascularly perfused rat stomach. The gastric postganglionic sympathetic nerves were electrically stimulated twice at 2.5 Hz for 1 min and test reagents were added during the second stimulation. The electrically evoked release of noradrenaline was augmented by tetraethylammonium and 4-aminopyridine (non-selective K+ channel blockers) and also by charybdotoxin (a blocker of big conductance Ca2+-activated K+ channel). On the other hand, apamin (a selective blocker of small conductance Ca2+-activated K+ channels) and glibenclamide (an ATP-activated K+ channel blocker) had no effect on the evoked noradrenaline release. Oxotremorine-induced inhibition of noradrenaline release was attenuated by tetraethylammonium and 4-aminopyridine, while the inhibition was not influenced by charybdotoxin, apamin, and glibenclamide. These results suggest that tetraethylammonium- and 4-aminopyridine-sensitive K+ channels (probably voltage-activated K+ channels) are involved in the muscarinic receptor mediated inhibition of noradrenaline release from the rat stomach. PMID- 15528843 TI - Effect of mexiletine on fenvalerate-induced nociceptive response in diabetic mice. AB - The effect of mexiletine on the nociceptive behavior induced by the intrathecal injection of fenvalerate, which predominantly activates tetrodotoxin-resistant (TTX-R) sodium channels, was studied in diabetic mice. The intrathecal injection of fenvalerate induced a characteristic behavioral syndrome that mainly consisted of reciprocal hind limb scratching directed toward caudal parts of the body and biting or licking of the hind legs in mice. The intensity of fenvalerate-induced nociceptive responses was significantly greater in diabetic mice than non diabetic mice. This fenvalerate-induced behavior was dose-dependently inhibited by mexiletine (3-30 mg/kg, i.p.). Intrathecal pretreatment with fenvalerate produced thermal hyperalgesia and allodynia in the tail-flick test in naive mice. Furthermore, mexiletine at doses of 10 and 30 mg/kg, i.p., dose-dependently and significantly reduced fenvalerate-induced thermal hyperalgesia and allodynia in the tail-flick test in naive mice. These present data suggest that i.p. pretreatment with mexiletine produced dose-dependent inhibition of fenvalerate induced hyperalgesia and allodynia in mice, especially diabetic mice. This effect may be, at least in part, mediated by the inhibition of TTX-R sodium channel mediated nociceptive transmission in the spinal cord. PMID- 15528844 TI - Experimental transmission of ovine herpesvirus-2 in sheep. AB - Transmission of ovine herpesvirus-2 (OvHV-2) in sheep via natural contact and nasal secretions was examined. OvHV-2-free lambs were produced by separating newborn lambs from their mothers within 5 days of birth and raising them in an isolation facility. Transmission experiments via natural contact were conducted by keeping OvHV-2-free lambs with OvHV-2-infected sheep of different ages. Six of the infected ewes in this experiment were pregnant and gave birth during the experimental period. OvHV-2 was not transmitted from the adult sheep, though viral DNA was consistently detected in their peripheral blood leukocytes (PBL). On the other hand, OvHV-2 was transmitted from recently infected lambs to sheep at 10 or 12 weeks after the onset of contact. In addition, we attempted the experimental transmission of OvHV-2 via nasal secretions, by transferring nasal washings from infected sheep to the nostrils of uninfected sheep. Sheep receiving the nasal washings from infected adult sheep maintained their negative status for 15 months, whereas sheep receiving nasal washings from recently infected lambs acquired OvHV-2 by 8 months. The results of these experiments support that OvHV-2 is more easily transmitted to negative sheep by recently infected lambs than by adult sheep. Further, it is supposed that the nasal cavity is a portal for entry and shedding of infectious OvHV-2 in sheep. PMID- 15528845 TI - High sensitivity of fibroblast cell lines derived from LEC rats to heat treatment. AB - A fibroblast cell line derived from LEC rat was approximately twofold more sensitive to heat treatment at 45 degrees C than were that from WKAH rat in terms of heating time required to attain 50% loss of survival in a colony forming assay. The present study was carried out for understanding the mechanism underlying the higher sensitivity of LEC rat cells to heat treatment. Although apoptosis was not found in WKAH rat cells, the percentages of apoptotic cells in LEC rat cells significantly increased after heat treatment. LEC rat cells showed significantly lower sensitivity in induction of cell death and apoptosis to ceramide, a lipid signaling molecule that is associated with heat-induced apoptosis, than did WKAH rat cells. SP600125, an inhibitor of JNK suppressed the induction of cell death in both heated LEC and WKAH rat cells, but SB203580, an inhibitor of p38 mapk, did not. The relative surviving fractions of heated LEC and WKAH rat cells in the presence of both SB203580 and SP600125 were higher than those of cells in the presence of SP600125 alone. The amounts of hsp70 protein in WKAH rat cells increased from 4 to 12 hr after heat treatment, but did not in LEC rat cells. These results suggest that higher thermosensitivity in the fibroblast cell line from LEC rat is due to low inducibility of hsp70 protein after heat treatment. PMID- 15528847 TI - Effect of lipopolysaccharide (LPS) injection on the immune responses of LPS sensitive mice. AB - The effect of lipopolysaccharide (LPS) on humoral and cell-mediated immunity was assessed using LPS-sensitive C3H/HeN mice. A single injection of LPS significantly decreased the anti-sheep red blood cells (SRBC) antibody titers, but not the number of anti-SRBC antibody producing spleen cells. In contrast, double LPS injection did not significantly decrease the anti-SRBC titers and even increased the number of anti-SRBC antibody producing spleen cells. Similarly, single LPS injection significantly suppressed the swelling of the footpad, but double LPS injection caused milder suppression. These results suggest that a difference in the level and timing of exposure to LPS may influence the immune response to infection or vaccination. PMID- 15528846 TI - Distribution of estrogen receptor alpha in the dominant follicles and corpus luteum at the three stages of estrous cycle in Japanese black cows. AB - Distribution of estrogen receptor alpha (ERalpha) in the dominant follicle (DF) and corpus luteum (CL) at the three stages of estrous cycle in Japanese Black cows was evaluated by means of immunohistochemistry. Ovarian dynamics were observed twice daily using ultrasonography until the ovariectomy performed on Day 7 (First group, n=3), Day 10 (Second group, n=3) and Day 18 (Third group, n=3) (Day 0=estrus). Expression of ERalpha represented by immunohistological staining intensity in cells was determined using a light microscope equipped with a digital camera. A tendency toward higher expression were observed in theca interna (TI) of DF when compared with those in mural granulosa cells (mGC), antral granulosa cells (aGC) and theca externa (TE). ERalpha expression in the Third group was lower than that in the First Group in mGC, and it was also lower than that in the second group in TE (P<0.05). ERalpha expression in luteal cells was higher than those in the stromal cells in CL. No significant difference of ERalpha expression was observed within luteal or stromal cells, except in the Second group in the luteal cells, in which significantly higher expressions than that in the Third group (P<0.05) were observed. The results showed that, 1) ERalpha was present in developing DF on Day 7, early regressing DF on Day 10 and preovulatory DF on Day 18, especially in the TI, and a few were localized in the mGC, and 2) ERalpha was highly expressed in the luteal cells and the expression decreased in combination with regression of CL. PMID- 15528848 TI - Enzymatic characterization of a cubilin-related serine proteinase from the hard tick Haemaphysalis longicornis. AB - In the present study, we performed enzymatic characterization of Haemaphysalis longicornis serine proteinase (HlSP) with a view to shed light on the mechanisms of blood digestion in the hard ticks. Escherichia coli-expressed recombinant HlSP (rHlSP) was shown to potently hydrolyze the synthetic substrates Bz-(DL)-Arg-pNA, Z-Ala-Ala-Leu-pNA and Suc-Ala-Ala-Ala-pNA and yielded an activity of 31.5, 88.2 and 18.3 mumol/min/mg protein, respectively at an optimum temperature of 25 degrees C. However, the enzyme showed little activity to hydrolyze the substrates Suc-Arg-Pro-Phe-His-Leu-Leu-Val-Tyr-MCA and Pyr-Phe-Leu-pNA. The optimum pH for the enzyme was shown to be 4.0 to 5.0. Several inhibitors such as antipain, leupeptin and phenylmethylsulfonyl fluoride (PMSF), specific for serine proteinase were shown to inhibit enzyme activity by 20-82%, while E-64 (specific for cysteine proteinases) and pepstatinA (specific for aspartic proteinases) had shown only little inhibitory effects on it. This is the first report on enzymatic characterization of a functional serine proteinase from the hard ticks. PMID- 15528849 TI - Enzyme-linked immunosorbent assay for bovine apolipoprotein A-IV. AB - The present report describes an enzyme-linked immunosorbent assay for bovine apolipoprotein (apo) A-IV. This assay was applied to the determination of its concentration and distribution in sera from cattle. The distribution of apoA-IV in lipoprotein fractions separated by ultracentrifugation was mostly recovered in the non-lipoprotein fractions (d>1.21 g/ml, 90%), but, in the case of gel filtration chromatography, apoA-IV was mainly eluted in HDL and non-lipoprotein fractions. The apoA-IV concentrations during early, mid- and late lactating stages in cows were significantly higher than during the nonlactating stage (p<0.05). From early to late lactating stages, the concentration of apoA-IV was unaltered. After 4 days of fasting, the concentration of plasma apoA-IV had decreased significantly (p<0.05) at days 3 and 4, and was returned to the basal level by 3 days of refeeding. These results suggested that the concentration of apoA-IV is modified by nutritional conditions. PMID- 15528850 TI - The effect of polysaccharides and carboxymethylcellulose combination to prevent intraperitoneal adhesion and abscess formation in a rat peritonitis model. AB - Polysaccharides isolated from fungi, Phellinus spp. is well-known material with anti-tumor and anti-inflammatory properties. We have assessed the adhesion- and abscess-reducing capacity of carboxymethylcellulose (CMC) and polysaccharides from Phellinus spp. combination in a rat peritonitis model. In 72 Sprague-Dawley rats, experimental peritonitis was induced by means of the cecal ligation and puncture model (CLP). After 24 hr, the abdomen was reopened and the ligated cecum was resected. Peritoneal fluid samples were taken for microbiological examination. Rats were randomly assigned to 6 groups: ringer lactate solution (RL group), polysaccharides from Phellinus gilvus (PG group) and Phellinus linteus (PL group), carboxymethylcellulose (CMC group), and their combinations (PG+CMC and PL+CMC groups). Adhesions and abscesses were noted at day 7 after CLP. RT-PCR assay for urokinase-type plasminogen activator (uPA), its cellular receptor (uPAR), and tumor necrosis factor (TNF)-alpha was performed to assess the cecal tissue. Microbiological examination showed polymicrobial bacterial peritonitis. Adhesion formation was significantly reduced in PG+CMC and PL+CMC groups (P<0.05). The incidence of abscesses was reduced in all treated groups except the RL group (P<0.05). uPA, uPAR, and TNF-alpha mRNA were highly expressed in the PG+CMC and PL+CMC groups, as compared to the RL group. We concluded that the combination of polysaccharides and CMC had significant adhesion- and abscess reducing effects compared with their single treatment and the effects may act by modifying the fibrinolytic capacity of uPA, uPAR and TNF-alpha produced from activated macrophages in a rat peritonitis model. PMID- 15528851 TI - Geographical variation of skull size and shape in various populations in the black giant squirrel. AB - We osteometrically examined the skulls of the black giant squirrel (Ratufa bicolor) from three mainland populations (M. Malayan Peninsula, V. South Vietnam, and B. Burma, India and North Thailand) and from two island populations (T. Tioman, and S. Sumatra Islands). The skull in the Malayan peninsula population was significantly smaller than that of the two other mainland populations. It is consistent with Bergmann's rule as shown in the gray-bellied squirrel. The two island populations did not show obvious differences in comparison with the Malayan population in many measurements. In the proportion analysis eliminating the size factor, the differences among populations were not easily confirmed and we concluded that the osteological characters peculiar to each population could not be shown in this species. The first and second principal component scores of M, S, and T populations were intermingled, whereas the V and B populations of V and B were not separated in the chart. We pointed out that the morphological differences were demonstrated between northern and southern groups of the Isthmus of Kra in the mainland populations, and that the two island populations did not show the island-isolation effect in comparison with the M population. The adaptational variation related to feeding and locomotion could not be confirmed among populations of the black giant squirrel as shown in the proportion analysis. PMID- 15528852 TI - Glutathion peroxidase and glucose-6-phosphate dehydrogenase activities in bovine blood and liver. AB - A total of 46 cattle, including 25 as control, 16 with glycogen degeneration and 5 with severe fatty degeneration were studied. Whole blood and liver tissue specimens were used to measure glutathione peroxidase (GSH-Px) and Glucose-6 Phosphate Dehydrogenase (G6PD) activities. The present study determined the value of these parameters in diagnosing glycogen and fatty degeneration in cattle from the point of the status of antioxidation and lipid peroxidation. The results showed a significant decrease in hepatic GSH-Px activity and a significant increase in hepatic G6PD activity in cases of fatty degeneration. On the other hand, there were no significant changes in erythrocytic and hepatic GSH-Px and G6PD activities in cases of glycogen degeneration. The results indicated lipoperoxidation process in the liver tissues increased in cases of fatty degeneration. Therefore, supplying animals suffering from fatty liver with sufficient quantities of nutrient antioxidants may be valuable when treatment is considered. PMID- 15528853 TI - Effects of the neutrophil elastase inhibitor (ONO-6818) on acetic acid induced colitis in Syrian hamsters. AB - Neutrophil elastase (NE) released from neutrophils during inflammation is related to tissue disturbance and organ failure. We investigated the effects of an orally active NE inhibitor, ONO-6818, on acetic acid induced colitis in Syrian hamsters. The ulcer area, hemoglobin level in the colonic lumen, NE activity, and tissue myeloperoxidase (MPO) activity in the colitis control animals were significantly increased compared to the normal control ones. Either oral or subcutaneous treatment with ONO-6818 had significant inhibitory effects on the ulcer area, hemoglobin level and NE activity in the colonic lumen, but ONO-6818 did not have a significant inhibitory effect on tissue MPO activity. We conclude that NE is closely related to the development of inflammation in acetic acid-induced colitis in Syrian hamsters and that the condition is improved by the inhibition of NE. PMID- 15528854 TI - Geographical variation of the skull of the lesser mouse deer. AB - We examined the geographical variation of the skull size and shape of the lesser mouse deer (Tragulus javanicus) from Laos, Thailand, Peninsular Malaysia, Sumatra, Java, Borneo, Langkawi and some Islands of Tenasserim in Myanmar. Although the influence of the climatic condition on skull size was not confirmed in the mainland populations, the skull became rostro-caudally longer in the populations of Tenasserim and Sumatra because of island isolation effect. The skull size was classified into the following three clusters of localities from the matrix of Q-mode correlation coefficients: 1) Langkawi and Tenasserim, 2) Laos and Thailand, 3) Sumatra and Borneo. The skulls in the population of Java belong to the cluster of Langkawi and Tenasserim in male, however were morphologically similar to those in the cluster of Borneo and Sumatra. The canonical discriminant analysis pointed out that the Laos and Tenasserim populations were separated from the other ones and that the populations of Sumatra, Java and Borneo were intermingled each other. PMID- 15528855 TI - Modulation by sphingosine of phosphorylation of substrate proteins by protein kinase C in nuclei from cow mammary gland. AB - Protein kinase C (PKC) is an enzyme activated by diacylglycerols such as 1-oleoyl 2-acetyl-sn-glycerol (OAG), phospholipids (in particular phosphatidylserine; PS) and Ca2+, which regulate a wide variety of intracellular functions by phosphorylating multiple substrate proteins and enzymes. The effect of sphingosine, the backbone moiety of sphingolipids, on PKC activity and phosphorylation of endogenous proteins catalyzed by PKC was investigated in nuclei of cow mammary gland. Sphingosine inhibited nuclear PKC activity when lysine-rich histone was used as the substrate. The sphingosine inhibition of the PKC activity was reversed by the excess addition of PS, but not by OAG or Ca2+. Several nuclear proteins, including 56-kDa, 43-kDa, 38-kDa and 36-kDa proteins, were shown to be substrates for PKC. Of the substrate proteins, the 38-kDa and 36 kDa proteins were identified as annexin I, the Ca2+/phospholipid-binding protein; the 56-kDa and 43-kDa proteins have not yet been identified. Sphingosine inhibited phosphorylation of the 56-kDa protein and the 36-kDa annexin I, whereas it enhanced that of the 43-kDa protein. The 38-kDa annexin I species was unaffected by sphingosine. As with the PKC activity, inhibition by sphingosine of phosphorylation of the 56-kDa protein and 36-kDa annexin I was reversed by the excess addition of PS, but not by OAG or Ca2+. In addition, by the excess addition of PS and not by OAG or Ca2+, the sphingosine-enhanced phosphorylation of the 43-kDa protein was reversed and returned to near the level in the absence of sphingosine. It is suggested that sphingosine is involved in the regulation of PKC-dependent phosphorylation in the nucleus by modulating the association of PKC or its substrates, particularly annexin I, with membrane phospholipids in cow mammary gland. PMID- 15528856 TI - Phylogenetic analysis of Fusobacterium necrophorum, Fusobacterium varium and Fusobacterium nucleatum based on gyrB gene sequences. AB - The nucleotide sequences of the DNA gyrase B subunit gene (gyrB) of Fusobacterium necrophorum subsp. necrophorum, F. necrophorum subsp. funduliforme and F. varium were determined and analyzed together with those of F. nucleatum subsp. nucleatum and F. nucleatum subsp. vincentii. On the phylogenetic tree constructed, the strains of each fusobacterial species formed distinct clusters with deep sublines. The degree of sequence similarity within each cluster was 93.2% or more, whereas similarities between clusters ranged from 70.1 to 72.7%. These clusters were recovered with 100% bootstrap probabilities and are in very good agreement with the species of Fusobacterium. These data suggest that gyrB is an accurate genealogical marker for the classification of the fusobacterial taxa considered in this study. PMID- 15528857 TI - Clinical features of skin lesions in rabbit syphilis: a retrospective study of 63 cases (1999-2003). AB - Skin lesions in rabbit syphilis are usually diagnostic, but it is occasionally difficult to differentiate these lesions from those of other skin diseases. Skin lesions in 63 cases of rabbit syphilis were analyzed for early and accurate diagnosis. Lesions were found most frequently around the nose (55 cases) followed by the genitalia (22), lips (20), eyelids (12), and anus (10). Sneezing was observed in 33% of cases with nasal lesions. In cases of maternally acquired infection, lesions could be initially found mainly on the face. Rabbits should be examined carefully not only for facial lesions, but also for lesions of the genitalia and anus, locations easily overlooked. PMID- 15528858 TI - Complete cDNA sequence and mRNA expression of dog preproendothelin-3. AB - The full-length cDNA of dog preproendothelin-3 (PPET3) was cloned from lung tissue using RT-PCR and rapid amplification of cDNA ends. Aside from the poly (A) tail, the full-length cDNA was 1976 bp. A polyadenylation signal sequence and one copy of a consensus sequence, ATTTA, which is related to mRNA turnover, was found in the 3' noncoding region. The cDNA had a 594-bp open reading frame encoding a 198-amino acid polypeptide. Regions corresponding to a bioactive mature ET3 peptide, an intermediate form known as big-ET3, and an ET3-like peptide were observed in dog PPET3. Expression of PPET3 mRNA was detected throughout the organs examined, which included heart, lung, liver, kidney, spleen, stomach, pancreas, duodenum, colon, uterus, ovary and testis. PMID- 15528859 TI - Molecular cloning and sequencing of feline CD7. AB - Human CD7 is one of the earliest molecules to appear in T cell development. In this study, putative feline CD7 cDNA was identified based on its similarities with human and mouse CD7 genes. The feline CD7 cDNA contained an open reading frame consisting of 630 nucleotides. The amino acid sequence of feline CD7 had 47.7% identity with that of human CD7, and 52.9% with that of mouse CD7. In addition, the feline CD7 protein fused with histidine tag was expressed in 293T cells. The expression was confirmed by indirect immunofluorescence assay. PMID- 15528860 TI - Morphometrical analysis of the kidney from nonobese diabetic (NOD) mice in the non-diabetic stage. AB - The kidneys of non-diabetic NOD and wild type ICR mice were examined morphometrically at 3 and 6 months of age. Kidney weights and diameter of renal corpuscles of non-diabetic NOD mice were less than those of ICR mice. No lesions were observed in glomeruli or uriniferous tubules. Renin-positive areas were more common in NOD mice than in ICR mice, but no differences were detected in the Western blot analyses. PMID- 15528861 TI - Reactivity of anti-Nipah virus monoclonal antibodies to formalin-fixed, paraffin embedded lung tissues from experimental Nipah and Hendra virus infections. AB - The immunohistochemical reactivity of seven clones of mouse monoclonal antibodies raised to Nipah virus antigens were investigated using formalin-fixed, paraffin embedded porcine and equine lung tissues from experimental Nipah and Hendra virus infection, respectively. Either microwave irradiation or enzymatic digestion effectively unmasked the viral antigens in formalin-fixed, paraffin-embedded tissue sections. Four clones showed positive reaction to both Nipah virus infected porcine lung tissue and Hendra virus-infected equine lung tissue. Two clones (11F6 and 13A5) reacted with Nipah virus-infected porcine lung tissue, but not with Hendra virus-infected equine lung tissue. These Nipah virus-specific monoclonal antibodies may therefore be useful for immunohistological diagnosis of Nipah virus infection and for further research on Nipah virus pathogenesis. PMID- 15528862 TI - Susceptibilities against bovine lactoferrin with microorganisms isolated from mastitic milk. AB - Antibacterial effects of bovine lactoferrin were studied in vitro against microorganisms isolated from mastitic milk in Tokachi area, Hokkaido, Japan. Microorganisms isolated were Escherichia coli (11 isolates), Klebsiella pneumoniae (5 isolates), enterococci (8 isolates), Staphylococcus aureus (10 isolates), coagulase negative staphylococci (CNS, 13 isolates), streptococci (11 isolates), Prototheca zopfii (7 isolates) and yeast-like fungi (9 isolates). Lactoferrin has been known as a multifunctional protein and its antimicrobial effect is one of the most essential function of it. In order to compare their susceptibilities against lactoferrin, the minimal inhibitory concentration values were estimated by a microplate assay method using 96-well microplate, which involved measuring the optical density of the cultures. Prototheca zopfii was highly sensitive to bovine lactoferrin and complete inhibition of this microorganism was observed even at the low concentration of 7 mug/ml. On the other hand, E. coli and enterococci showed resistance against lactoferrin action and staphylococci showed strain-dependent resistance. PMID- 15528863 TI - Genetic characterization of rabies viruses isolated from frugivorous bat (Artibeus spp.) in Brazil. AB - In Latin America, rabies cases related to frugivorous bats have been reported since 1930's. Recently, two viruses isolated from Artibeus lituratus were proved to be vampire bat variants by monoclonal antibodies panels [2], but their genetic information is not well known. In this report, four rabies viruses were isolated from frugivorous bats (Artibeus spp.) in Brazil and their nucleoprotein gene sequences were determined. These isolates were found to be genotype 1 of lyssavirus and showed the maximum nucleotide sequence homology of 97.6-99.4% with vampire bat-related viruses in Brazil [6]. These results indicate that the Brazilian frugivorous bat rabies viruses in this study are closely related to vampire bat-related viruses that play a main role in rabies virus transmission to livestock in Brazil. PMID- 15528864 TI - Morphogenesis of the olfactory pit in a flatfish, barfin flounder (Verasper moseri). AB - Morphogenesis of the olfactory pit (OP), olfactory lamella (OL) and olfactory epithelium (OE) was examined by scanning electron and light microscopy in the barfin flounder (Verasper moseri). At day 0 after hatch, the OP was already formed. At day 14, the cellular differentiation of the OE was prominent. At day 42, the OP became a cavity by the formation of its roof. At day 56, the first OL extended remarkably and was lined with the OE on both sides. The OL increased in number with development. These findings suggest that the OE is functionally active at day 14. The formation of the OL in the OP may be initiated by the stimulus when the barfin flounder touched at the bottom of the sea. PMID- 15528865 TI - Intestinal adenocarcinoma with pancreas and lymph node metastases in a captive cotton-top tamarin (Saguinus oedipus). AB - A case of intestinal adenocarcinoma with metastases to the pancreas and regional lymph node was found in a 9-year-old, captive female cotton-top tamarin (Saguinus oedipus) with intermittent diarrhea. At necropsy, the tumor mass was located in the ileo-cecal junction causing circumferential thickening and annular stenosis. Microscopically, the lesions at primary and metastatic sites showed typical features of mucinous adenocarcinoma as seen in humans, including intracellular and extracellular mucin production and characteristic appearance of a signet ring of the tumor cells. The diagnosis was confirmed by histological evaluation, positive cytokeratin immunostain, and mucin production demonstrated by PAS and Alcian blue stain. It is speculated that the development of intestinal carcinoma was partly attributable to the excessive absorption of a diet of refined food, unbalanced nutrition, and the nature of these animals to develop stress easily. PMID- 15528866 TI - Intestinal microflora in 45 crows in Ueno Zoo and the in vitro susceptibilities of 29 Escherichia coli isolates to 14 antimicrobial agents. AB - Microorganisms from 45 jungle crows (Corvus macrorhynchos) captured from July to December 2002 at Ueno Zoo, Tokyo were identified as Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae, Enterobacter agglomerans, Pseudomonas maltophila, Staphylococcus spp., Micrococcus spp., and Streptococcus spp. E. coli showed the highest rate of isolation (21.6%). In an in vitro susceptibility test for 29 isolates of E. coli to 14 antimicrobial agents, all the isolates were resistant to penicillin G, vancomycin, erythromycin, lincomycin, bicozamycin, sulfadimethoxine, and olaquindox. Several isolates of them were also resistant to tetracycline, oxytetracycline, streptomycin, chloramphenicol, and ampicillin. Twenty-nine isolates were divided into 19 serogroups and the most frequently identified serogroups were O8, O114 and O144, which showed the same multidrug resistant patterns. PMID- 15528867 TI - Mixed feed containing dextran improves milk production of holstein dairy cows. AB - Total 37 Holstein daily cows (body weight: 631.76 +/- 18.45 kg, age: 5.47 +/- 1.94 years, parturition: 3.71 +/- 1.76 times) which became pregnant and gave birth to calves in the same season and lactated continuously were selected for this study. They were randomly divided into two groups: Group A-control, Group B fed with 30 g/head/day of mixed feed containing supplemental dextran for one year from October 2001. After supplementation of the mixed feed, milk yields and components (fat, protein and solid non-fat) of Group B were compared with those of Group A in the 8th, 10th and 11th months (May, July and August of 2002). Milk yields of Group B were greater than the yields of Group A. In particular, there was a significant difference (p<0.001) between these groups in the July and August values. Milk components of Group B slightly differed from those of Group A before the supplementation, but after the supplementation, concentrations and total amounts of fat, protein and solid non-fat significantly increased more in Group B than in Group A. Thus mixed feed containing dextran can increase the milk production of Holstein dairy cows in the hot season. PMID- 15528868 TI - Use of monoclonal antibodies for analyses of Coxiella burnetii major antigens. AB - Monoclonal antibodies (MAbs) to major antigens of Coxiella burnetii were produced. Some of the MAbs to a 62-kDa protein antigen, peptidoglycan protein complex and lipopolysaccharide (LPS) O-chains reacted with other bacteria whereas none of the MAbs to outer membrane proteins and LPS outer-core did. The LPS outer core and OMPs may be useful antigens for specifically detecting antibodies to C. burnetii. PMID- 15528869 TI - Unique amino acid polymorphisms of PrP genes in Mongolian sheep breeds. AB - To characterize amino acid polymorphisms of sheep prion protein (PrP) gene, DNA from 740 sheep of nine breeds raised in Mongolia was isolated and analyzed. A total of 16 genotypes and seven allelic variants of the PrP gene at codons 112, 136, 154, and 171 were found. The MARQ/MARQ genotype associated with susceptibility to scrapie was found in 82.6% of the sheep while the MARR/MARR genotype associated with resistance to scrapie was found in 1.8% of the sheep. The polymorphisms of valine and serine at codon 127, and leucine and arginine at codon 189 were detected in eight Mongolian sheep breeds, suggesting that these polymorphisms are a common feature among Mongolian sheep breeds. PMID- 15528870 TI - Involvement of thalamic paraventricular nucleus in the anticipatory reaction under food restriction in the rat. AB - To investigate which brain regions are involved in the anticipatory activity in rats restricted feeding for 2 hr, we examined c-Fos expression before and after feeding. Only the thalamic paraventricular nucleus (tPVN) showed c-Fos expression before feeding than after feeding. After the anticipatory locomotor activity rhythm was established, lesioning the tPVN attenuated this rhythm, but not the light-dark entrained rhythm. The anticipatory increase of blood corticosterone levels was not established in long-term tPVN-lesioned rats. These results suggest that the tPVN is involved in the expression of anticipatory reactions under a food-restricted regimen. PMID- 15528871 TI - Chloramphenicol treatment for rabbit syphilis. AB - Penicillin, the recommended treatment for rabbit syphilis, sometimes induces adverse effects. The efficacy of oral chloramphenicol was evaluated in 39 cases of rabbit syphilis to establish a safe and efficient treatment for this disease in companion rabbits. All cases clinically improved and recovered promptly. Fourteen of 39 cases (35.9%) relapsed, but most remained chloramphenicol sensitive. Since safety take priority over efficacy in treating syphilis in companion rabbits, chloramphenicol should be chosen as a first-line agent, as a general rule. Three-week administration of chloramphenicol may be adequate at the initial onset of disease. When relapse occurs repeatedly or the rabbit owner cannot administer the medicine adequately, treatment with penicillin should be considered. PMID- 15528872 TI - Thiazole orange positive platelets in a dog with Evans' syndrome. AB - We examined transition for the percentage of reticulated platelets (RP%) and platelet count in a canine case of Evans' syndrome. The result demonstrated that measurement of the RP% can be useful in evaluating platelet production in the bone marrow and response to treatment. PMID- 15528873 TI - Immunohistochemical study of osteopontin in the spinal cords of rats with clip compression injury. AB - Expression of osteopontin (OPN) was investigated in the spinal cords of rats with clip compression injury. Western blot analysis demonstrated that OPN protein increased significantly in the spinal cord during the early stages after injury. The increased expression of OPN was partially paralleled by that of proliferating cell nuclear antigen (PCNA). Immunohistochemical staining showed that OPN was expressed in proliferating activated microglia/macrophages in core lesions and in some astrocytes at the periphery of lesions. These results indicate that expression of OPN protein increases mainly in activated microglia/macrophages after spinal cord injury, suggesting that OPN is related to cell proliferation during the early stages after injury, probably leading to tissue remodeling. PMID- 15528874 TI - Effects of combination treatment with losartan and trandolapril on office and ambulatory blood pressures in non-diabetic renal disease: a COOPERATE-ABP substudy. AB - BACKGROUND: In the COOPERATE trial, the combination treatment of the angiotensin II receptor blocker losartan and the angiotensin-converting-enzyme inhibitor trandolapril significantly retarded progression of non-diabetic kidney disease compared with each monotherapy. The benefit could be greatly attributable to the potent reduction of proteinuria, because the three treatment groups showed the same reductions of office blood pressure (OBP). Ambulatory blood pressure (ABP) is reported to be better than OBP in predicting progression of kidney disease. METHODS: Ninety-two patients enrolled in the COOPERATE trial underwent 24-hour ABP monitoring at randomization and at month 6, year 1, year 2 and year 3 on randomized treatment. RESULTS: Both OBP and ABP were similarly reduced among the three groups at all measurement points (p = NS) and throughout the whole study period (p = NS). No significant correlation between the change in 24-hour ABP and the change in proteinuria was seen (p = NS). A Cox-multivariable analysis showed that covariates affecting the renal outcomes (a doubling serum-Cr level and/or end-stage renal failure) were the change in proteinuria (hazard ratio 0.49, 95% CI 0.34-0.78, p = 0.01) and treatments (0.58, 0.45-0.99, 0.03), but not 24-hour ABP (0.98, 0.89-2.01, 0.17). CONCLUSION: The better renoprotective effect of the combination treatment is attributed to BP-independent mechanisms by more complete renin-angiotensin system blockade. PMID- 15528876 TI - The 'myth' of asphyxia and hypoxia-ischemia as primary causes of necrotizing enterocolitis. PMID- 15528875 TI - Insulin-like growth factor attenuates apoptosis and mucosal damage in hypoxia/reoxygenation-induced intestinal injury. AB - OBJECTIVE: Necrotizing enterocolitis (NEC) is a potentially lethal disease among premature infants. The aim of the present study was to investigate whether hypoxia-reoxygenation (H/R)-induced intestinal injury was due to increased apoptosis of the intestinal mucosa in young mice and whether pre-treatment of the animals with recombinant human insulin-like growth factor-I (IGF-I), a known anti apoptotic factor, could protect the intestinal cells from H/R-induced apoptosis or intestinal injury. STUDY DESIGN: Young mice were divided into three groups: group 1 mice (H/R) were hypoxia-reoxygenation; group 2 mice (H/R + IGF-I) were treated with recombinant human IGF-I by intraperitoneal injection (1 mug/g b.w. once daily) for 7 days, and group 3 mice served as control. Hypoxia was induced by placing young mice in a Plexiglas chamber consisting of 10% oxygen for 60 min. After hypoxia, the young mice were reoxygenated for 10 min with 100% oxygen. Intestinal generation of substances reactive to thiobarbituric acid (TBARS) and active caspase-3 were measured in H/R-induced intestinal injury. RESULTS: Increased numbers of apoptotic cells (apoptotic index) across the villi in young mice subjected to H/R were observed with the TUNEL reaction whereas few apoptotic cells existed in the control animals. In addition, H/R-induced intestinal damage in the H/R + IGF-I group was greatly attenuated, with necrosis limited partially to the mucosa. Tissue-active caspase-3 levels in the H/R group were found to be significantly higher when compared with that of the H/R + IGF-I group of mice and control. However, TBARS concentrations in the intestine were similar in H/R groups when compared to the intestine of control animals. CONCLUSION: The present study suggests that both necrosis and apoptosis, via mechanisms occurring due to oxygen-derived free radicals and activation of caspase-3, play a role in the pathogenesis of H/R-induced bowel injury. We also show that IGF-I protect intestinal mucosa from necrosis and apoptosis from intestinal H/R injury. PMID- 15528877 TI - Effect of granulocyte and monocyte adsorption apheresis on urinary albumin excretion and plasma endothelin-1 concentration in patients with active ulcerative colitis. AB - BACKGROUND/AIM: Increases in microalbuminuria and endothelin (ET-1) are involved in the development of ulcerative colitis (UC) and in its progress. Because granulocyte and monocyte adsorption apheresis has proven to be useful in the treatment of UC, we examined whether urinary albumin excretion and plasma ET-1 concentrations are altered and whether granulocyte and monocyte adsorption apheresis affects the concentrations of these two factors in patients with active UC. METHODS: Twenty patients with active UC and 20 age-matched healthy volunteers (our hospital staffs) were included in this study. UC patients were randomly divided into two treatment groups: a granulocyte and monocyte adsorption treatment group (n = 10) and a conventional treatment group (n = 10). The urine albumin/creatinine ratio, plasma ET-1 concentration and tumor necrosis factor (TNF)-alpha were determined before and after treatment and compared between 2 treatment groups. The 10 adsorption treatment patients underwent 5 consecutive weekly apheresis sessions, each of 60 min duration at a flow rate of 30 ml/min. RESULTS: The urine albumin/creatinine ratio in UC patients (6.4 +/- 2.2 mg/mmol) were higher than that in healthy subjects (1.0 +/- 0.7 mg/mmol, p < 0.01). In addition, the plasma ET-1 level in UC patients (3.5 +/-1.5 pg/ml) was higher than that in healthy subjects (0.8 +/- 0.4 pg/ml, p < 0.01). Plasma TNF-alpha was detected in UC patients (18.8 +/- 8.4 pg/ml), but not in healthy subjects. The urine albumin/creatinine ratio was highly correlated with the plasma ET-1 level (r = 0.62; p < 0.01) and plasma TNF-a level (r = 0.66, p < 0.01). Granulocyte and monocyte adsorption apheresis reduced the urine albumin/ creatinine ratio from 6.6 +/- 2.4 to 1.8 +/- 0.6 mg/mmol (p < 0.01), reduced the plasma ET-1 level from 3.7 +/- 1.6 to 1.4 +/- 0.6 pg/ml (p < 0.05) and reduced the plasma TNF-alpha from 19.2 +/- 8.6 to 3.8 +/- 1.2 pg/ml (p < 0.01). Conventional treatment did not affect these factors. CONCLUSION: Our data suggest that increases in the urine albumin/creatinine ratio, ET-1 and TNF-alpha play an important role in active UC and that granulocyte and monocyte adsorption apheresis is effective in ameliorating such increases. PMID- 15528878 TI - Potential confounding by intermediate phenotypes in studies of the genetics of ischaemic stroke. AB - BACKGROUND: Family history (FHx) of stroke is perceived to be an important risk factor for ischaemic stroke. However, there are several intermediate phenotypes that are often involved in the aetiology of ischaemic stroke and that have a substantial genetic component themselves. We studied FHx of ischaemic heart disease (IHD), hypertension (HTN) and diabetes mellitus (DM) as risk factors for ischaemic stroke. METHODS: We performed a systematic review of case-control and cohort studies reporting on FHx(IHD), FHx(HTN) or FHx(DM) as risk factors for stroke using bibliographic databases, and by hand searching reference lists and journals. Odds ratios of FHx as a risk factor for stroke were calculated within individual studies. We included unpublished data from two Oxfordshire population based studies to assess effects on subtypes of ischaemic stroke. RESULTS: We identified 54 studies that investigated the odds of stroke conferred by a positive FHx, 24 of which reported data on FHx of one or more intermediate phenotypes in addition to FHx of stroke. Most studies reported an increased frequency of FHx(IHD) and FHx(HTN) in stroke patients versus controls. The association was significant in 6 out of 14 studies for FHx(IHD) and 4 out of 11 studies for FHx(HTN). In contrast, FHx(DM) was not associated with stroke. FHx(IHD) was particularly associated with large vessel strokes (OR 1.72, CI 1.3 2.2, p = 0.00004). CONCLUSIONS: FHx(IHD) and FHx(HTN) are both risk factors for stroke. It is likely that the apparent heritability of stroke is partly accounted for by heritability of HTN and large vessel atherosclerosis. Analyses of heritability of stroke and candidate gene studies should be adjusted accordingly. PMID- 15528879 TI - Burden of informal caregiving for stroke patients. Identification of caregivers at risk of adverse health effects. AB - BACKGROUND: We assessed the objective and subjective burden of caregiving for stroke patients and investigated which characteristics of the patient, the informal caregiver and the objective burden contribute most to subjective burden and to the condition of feeling substantially burdened. METHODS: We studied a sample of 151 stroke survivors and their primary informal caregivers. We collected data through patient and caregiver interviews 6 months after stroke. RESULTS: Both the level of subjective burden and the condition of feeling substantially burdened were associated with both caregiver's and patient's health related quality of life, patient's age, and the number of caregiving tasks performed. CONCLUSIONS: These conditions can be used in clinical practice to identify potentially vulnerable caregivers in need of support and at risk of adverse health effects. Monitoring stroke survivors as well as their family caregivers at discharge may help to prevent or alleviate caregiver burden. PMID- 15528880 TI - Risk factors for stroke among urbanised Indonesian women of reproductive age: a hospital-based case-control study. AB - BACKGROUND: Stroke is a major cause of death in South-East Asia, but few empirical data exist on its risks in Asian populations. METHODS: 235 cases and 682 age-matched controls of women of reproductive age (20-44 years) were recruited in 14 hospitals in Jakarta, Indonesia, between 1989 and 1993. Medical history was collected by a structured interview. In subsamples, glucosuria, serum cholesterol level and body mass index (BMI) were assessed. RESULTS: In these young and lean Muslim women, with few users of tobacco, alcohol or oral contraceptives, risk factors related to increased weight were strongly related to stroke occurrence. A history of hypertension or diabetes or increased serum cholesterol level showed odds ratios (ORs) of 13.9, 7.4 and 3.7, respectively. A BMI >27 (unadjusted for its potential consequences) caused an OR of 2.9. High social class and higher level of education (both OR 0.7) were associated with a lower risk of stroke, but levels of risk factors were higher in higher socio economic classes. CONCLUSION: The expected transition in lifestyle, characterised by a higher intake of calories and less physical activity, will increase stroke risks in Indonesian women. Increasing wealth should go together with raising levels of health education on nutrition and physical activity. PMID- 15528881 TI - Semi-intensive monitoring in acute stroke and long-term outcome. AB - BACKGROUND AND PURPOSE: Factors that determine the benefit of stroke units (SU) are unknown. The aim of our study was to analyze whether semi-intensive monitoring during the acute phase of stroke reduces mortality and dependency at long term. METHODS: We studied patients with an ischemic stroke or intracerebral hemorrhage, consecutively admitted to our SU within 24 h of symptoms onset. Based on bed availability, patients were allocated to either a conventional care stroke unit (C-SU, n = 209) or a semi-intensive stroke unit (SI-SU, n = 321) with continuous monitoring of cardiac, respiratory, metabolic and neurological functions during the first 72 h. Both groups were treated following the same medical and nursing protocols. Criteria for exclusion were patients with stupor/coma, previously dependent (Barthel score <85) and with TIA. Using logistic regression models, we analyzed the influence of semi-intensive care on mortality and dependency at one year. RESULTS: Baseline characteristics were similar between patients admitted to the SI-SU and the C-SU, except for a higher frequency of more severe stroke and intracerebral hemorrhage in the SI-SU. Twenty six percent of patients in the SI-SU and 4% in the C-SU were randomized in acute clinical trials (p < 0.01), and 61% and 39% were seen by a neurologist in less than 6 h from the onset of symptoms (p < 0.01). At 1 year, mortality and combined mortality and dependency were not significantly different between the two groups. However, due to the presence of a significant interaction between the type of unit and stroke severity, the OR of mortality for SI-SU allocation was 0.19 (95% CI, 0.07-0.54) in patients with severe stroke (CSS < or =4), whereas it was 0.64 (95% CI, 0.37-1.11) in those with mild-to-moderate stroke. CONCLUSIONS: This study suggests that semi-intensive monitoring in a stroke unit reduces mortality at 1 year in patients with severe stroke, with no influence over dependency. PMID- 15528882 TI - Delirium in acute subarachnoid haemorrhage. AB - BACKGROUND: Delirium may be a presenting feature in acute subarachnoid haemorrhage (SAH). The aim of this study was to investigate the risk factors for delirium in acute SAH and to analyse the relation between delirium and location and amount of haematic densities and hydrocephalus. METHODS: We assessed delirium in a sample of 68 consecutive patients with acute (< or =4 days) SAH (33 aneurysmal, 33 non-aneurysmal, including 9 with perimesencephalic haemorrhage), before aneurysmal treatment, using DSM-IV-R criteria and the Delirium Rating Scale (DRS). DRS scores were related to: (1) the total amount of haematic densities at 10 basal cisterns/fissures and in the 4 ventricles, using a validated rating scale, (2) the haematic densities in the prepontine cistern and the convexity of the brain and (3) hydrocephalus, using the bicaudate index, obtained from a review of admission CT scans. RESULTS: Eleven acute SAH patients presented with delirium. Older age (U = 316.5, p = 0.04), alertness disturbance (chi(2) = 5.1, p = 0.02, OR = 7.6, 95% CI = 1.5-37.3), aphasia (U = 61.5, p = 0.007) and a Hunt and Hess score >2 (U = 362.5, p = 0.02) were associated with delirium. Higher amounts of intraventricular haematic densities (chi(2) = 4.43, p = 0.04, U = 158, p = 0.001) and hydrocephalus (U = 215, p = 0.009) were also associated with higher DRS scores. Two delirious patients had basofrontal haematomas. CONCLUSIONS: Delirium was detected in 16% of acute SAH patients. Intraventricular bleeding, hydrocephalus and basofrontal haematomas contribute to the pathogenesis of delirium, through damage to anatomical networks subserving sustained attention, declarative memory and the expression of emotional behaviour. PMID- 15528883 TI - Trends in surgical and management outcomes in patients with aneurysmal subarachnoid hemorrhage in Izumo city, Japan, between 1980-1989 and 1990-1998. AB - BACKGROUND AND PURPOSE: The purpose of this community-based study was to evaluate temporal changes in surgical and management outcomes in patients with aneurysmal subarachnoid hemorrhage (SAH). METHODS: The subjects were 358 patients with aneurysmal SAH who were treated during the 19-year period from 1980 to 1998 in Izumo City, Japan. We compared data during the 9-year period 1990-1998 (period B; 188 patients) with those during the 10-year period 1980-1989 (period A; 170 patients). RESULTS: The proportion of patients 80 years of age or older or those with World Federation of Neurosurgical Societies grade V increased significantly (period A, 5 and 25%; period B, 18 and 35%, respectively). The operability rate did not change for patients 69 years of age or younger, whereas it increased significantly for those 70-79 years of age (period A, 48%; period B, 72%). The 6 month and 2-year case fatality rates in surgically treated patients decreased significantly (period A, 12 and 20%; period B, 2 and 8%, respectively), whereas they were virtually unchanged for overall management (period A, 41 and 46%; period B, 38 and 42%, respectively). In patients who underwent surgery, the incidence of permanent symptomatic vasospasm decreased from 21% during period A to 11% during period B, and there was no death from vasospasm in the later period. However, no significant difference was found in the functional outcome between the two periods, regardless of whether surgery was performed. The most important determinants of 6-month and 2-year survival rates were grade on admission, rebleeding and the site of the ruptured aneurysms. Age was also a significant predictor of the 6-month case fatality rate. CONCLUSIONS: For patients with SAH who underwent surgery, there were trends towards decreases in the case fatality rate and in the incidence of permanent symptomatic vasospasm. Nevertheless, the overall management outcome was still unsatisfactory, mainly because of increasing numbers of very elderly and/or high-risk patients. . PMID- 15528884 TI - Prothrombin 20210A and oral contraceptive use as risk factors for cerebral venous thrombosis. AB - BACKGROUND: This study investigates the association between cerebral vein thrombosis (CVT) and the mutations FV 1691A (factor V Leiden), PT 20210A and MTHFR 677TT and acquired factors including oral contraceptive (OC) use. METHODS: 26 patients (21 females) and 217 healthy controls (134 females) were studied. Multiple regression analysis was performed. RESULTS: The frequency of the three mutations in cases and controls were: PT 20210A, 23 versus 1%, odds ratio (OR) 21.40 (95% CI 4.29-118.75), p < 0.001; FV 1691A, 8 versus 1%, OR 5.94 (95% CI 0.66-46.9); MTHFR 677TT, 4 versus 7%, OR 0.54 (95% CI 0.03-4.08). OC use was more frequent in female patients over 14 years old than in controls (84 vs. 40%, OR 8.15, 95% CI 2.09-37.13, p < 0.001). The model that best explained the thrombotic risk included PT 20210A and OC use. CONCLUSIONS: PT 20210A and OC use are the main thrombophilic risk factors predisposing to CVT and should be routinely investigated in patients with this disease. PMID- 15528885 TI - Lumbar puncture and dural sinus thrombosis--a causal or casual association? AB - BACKGROUND: A few cases of cerebral venous thrombosis (CVT) were reported after a lumbar puncture (LP), suggesting a causal association. The purpose of our study was to document that LP might predispose to CVT by decreasing blood flow velocities (BFV) in veins or dural sinus. METHODS: We performed a transcranial Doppler ultrasound study to register the mean BFV of the straight sinus (SS) before, during and after LP. RESULTS: Thirteen patients were studied. LP induced a decrease of 47% of mean BFV in the SS. The mean decrease of BFV was significant immediately at the end (p = 0.003), 30 min after (p = 0.015) and more than 6 h after LP (p = 0.008). CONCLUSIONS: LP induced a sustained decrease of mean BFV in the SS. The decrease of venous blood flow is a possible mechanism contributing to the occurrence of CVT. PMID- 15528886 TI - Design, baseline characteristics and carotid intima-media thickness reproducibility in the PARC study. AB - BACKGROUND: Intima-media thickness (IMT) is associated with an increased risk of cardiovascular and cerebral ischemic events, but its correlation with the absolute cardiovascular risk is not known in large populations. The Paroi Arterielle et Risque Cardiovasculaire (PARC) Study is an epidemiological study designed to correlate conventional assessment of cardiovascular risk with the mean IMT of the common carotid. METHODS: In the PARC study, 6,416 subjects were enrolled, including 80.7% subjects with cardiovascular risk factors and 19.3% without. A specific methodology was designed to harmonize the acquisition and processing of data at the 283 centers. Interreader agreement on image quality and IMT measurement of the common carotid artery (CCAIMT) was assessed from a random sample of 10% of the PARC study population. RESULTS: The intraclass correlation coefficient was 0.98 (95% CI 0.966-0.985), and the accuracy was high (standard deviation of the error measurement: 0.0185 mm). CONCLUSIONS: The reproducibility of the measurements assessed by means of the intraclass correlation coefficient and the accuracy of the CCAIMT measurement obtained in the PARC Study demonstrate the feasibility of large multicenter studies of IMT measurement. PMID- 15528887 TI - Butyrate increases apoptosis induced by different antineoplastic drugs in monocytic leukemia cells. AB - BACKGROUND: Apoptosis is an essential form of cell death, the failure of which can lead to cancer development. Cancer including leukemia is usually treated with chemotherapeutic drugs that can be effective, but frequently problems are encountered that impair the success of the treatment. Butyrate is a short-chain fatty acid that can have many effects on different cells, including apoptosis. METHODS: The effect of a combination treatment with butyrate and antineoplastic agents Ara-C, etoposide and vincristine is evaluate on the leukemic cell line THP 1. RESULTS: We show that butyrate increased apoptosis induced by the three agents as seen by measurement of DNA content, annexin exposure and morphological characteristics. We also demonstrate that the process of apoptosis induced by butyrate and chemotherapeutic drugs involves the participation of caspases and induced activation of caspase-3, -8 and -9. CONCLUSIONS: We believe that butyrate could be a promising therapeutic agent for the treatment of leukemia in combination with other antineoplastic drugs. PMID- 15528888 TI - Uniform potent activity of the antiproliferative metal chelate trans bis(resorcylaldoximato)copper(II) against a large panel of human tumor cell lines in vitro. AB - BACKGROUND: The copper chelate trans-bis(salicylaldoximato)copper(II), CuSAO2, is a powerful antitumor agent in vivo, being capable of drastically increasing the life span of mice bearing Ehrlich ascites carcinoma, and has in many cases even a curative effect. This compound as well as some congeners, most notably the 4 hydroxylated analogue trans-bis(resorcylaldoximato)copper(II), CuRES2, also have potent antiproliferative activity against tumor cells in vitro. METHODS: CuRES2 was tested in vitro against a panel of 37 different types of human tumor cells (leukemias, small cell and non-small cell lung tumors, melanomas, as well as colon, breast, central nervous system, ovarian and renal tumors). The cells were incubated on microculture plates for 6 days, and the amount of viable cells was determined with the agent XTT, whose metabolic reduction gives a colored formazan compound. RESULTS: CuRES2 inhibited the proliferation of all the 37 cell lines studied. The IC50 values ranged between 0.575 and 8.57 microg/ml and the IC90 values between 1.74 and 20.6 microg/ml. Thus, it had distinct and very potent activity against all the cell lines studied, and the differences between the susceptibilities of different tumor types and cell lines were not very large. DISCUSSION: Clearly, CuRES2 is active against all the tumor cell lines studied. None of the cell lines (tumor types) demonstrates a significantly higher or lower susceptibility, but some trends appear to exist. Thus, most (but not all) of the leukemia and melanoma cell lines, the two ovarian cancer lines and one small cell lung cancer and one CNS cancer cell line were somewhat more susceptible than the cell lines on average. The results obtained for the melanomas are of special interest, since CuRES2 (and even more so CuSAO2) have very low aqueous solubilities. In cutaneous melanomas, their administration might, however, be very easy, since both compounds are soluble in dimethyl sulfoxide and could thus easily be administered topically in this solvent whose ability to be absorbed through the skin is well known. CONCLUSION: The present results on human tumor cell lines are in line with previous observations that CuSAO2 and CuRES2 essentially totally inhibit the proliferation of leukemia L1210 and Ehrlich ascites carcinoma cells in vitro in a concentration as low as approximately 5 microg/ml. PMID- 15528889 TI - Characterization of an active pharmaceutical ingredient by its dissolution properties: amoxicillin trihydrate as a model drug. AB - To characterize the dissolution of particles, a dissolution rate coefficient alpha, consisting of a geometric factor gamma and material constant mu, was introduced. The impact of the particle geometry on the initial rate of dissolution was assessed for spherical, cubic and tetrahedral particles. Additionally, a description of dissolution of samples containing multiple populations of particles was derived. A two-population model was employed to characterize sieved fractions of amoxicillin trihydrate. The investigation, using factor analysis, of the influence of dose, particle size and agitation intensity on the dissolution rate coefficient alpha indicated the presence of disintegrating agglomerates in larger particles. Based on the dissolution characteristics of particular particle size fractions and on the determined particle size distribution, the dissolution profile of the mixed sample containing nine sieved fractions in size was successfully predicted. Finally, the limitations of the film theory are discussed in light of the multiple-population dissolution theory. PMID- 15528890 TI - The emergence of glycopeptide-resistant Enterococcus faecium in a university hospital in southwestern Greece. AB - BACKGROUND: Enterococci and especially glycopeptide-resistant strains (GRE) are widely distributed in the hospital environment, by acquiring resistance determinants and virulence factors. METHODS: The study included 48 GRE isolated during a 1-year period from different inpatients in a tertiary hospital in southwestern Greece. Antibiotic susceptibility was determined by the Etest, and the presence of resistance and virulence genes was shown by PCR. Clonal types were identified by pulsed-field gel electrophoresis of SmaI DNA digests. RESULTS: All GRE were multi-resistant of the VanA phenotype, verified by the detection of the gene by PCR. Two major clones were distributed in all hospital wards. The majority of the strains (46 of 48) harbored the esp gene, while 27 GRE expressed also the gelE and/or as genes. CONCLUSIONS: The spread of two clones expressing the vanA gene and virulence factors were responsible for the emergence of GRE in the University Hospital of Patras. PMID- 15528891 TI - Vancomycin tolerance in enterococci. AB - BACKGROUND: Tolerance can be defined as the ability of bacteria to grow in the presence of high concentrations of bactericide antimicrobics, so that the killing action of the drug is avoided but the minimal inhibitory concentration (MIC) remains the same. We investigated vancomycin tolerance in the Enterococcus faecium and Enterococcus faecalis strains isolated from different clinical specimens. METHODS: Vancomycin was obtained from Sigma Chemical Co. We studied 100 enterococci strains. Fifty-six and 44 of Enterococcus strains were idendified as E. feacalis and E. faecium, respectively. To determine MICs and minimal bactericidal concentration (MBC), we inoculated strains from an overnight agar culture to Muller-Hinton broth and incubated them for 4-6 h at 37 degrees C with shaking to obtain a logarithmic phase culture. The inoculum was controlled by performing a colony count for each test. We determined MBC values and MBC/MIC ratios to study tolerance to vancomycin. Vancomycin tolerance was defined as a high MBC value and an MBC/MIC ratio > or =32. RESULTS: Fifty-six and 44 of the Enterococcus strains were identified as E. faecium and E. faecalis, respectively. Thirty-one E. faecium and 48 E. faecalis were found to be susceptible to vancomycin and these susceptible strains were included in this study. The MICs of susceptible strains ranged from < or =1 to 4 mg/l, the MBCs were > or =512 mg/l. Tolerance was detected in all E. faecalis and E. faecium strains. The standard E. faecalis 21913 strain also exhibited tolerance according to the high MBC value and the MBC/MIC ratio. We defined the tolerant strains as having no bactericidal effect and MBC/MIC > or =32. We found that a 100% tolerance was present in susceptible strains. CONCLUSIONS: One of the hypotheses for tolerance is that tolerant cells fail to mobilize or create the autolysins needed for enlargement and division. Our data suggests that tolerance may compromise glycopeptide therapy of serious enterococci infections. To add an aminoglycoside to the glycopeptide therapy unless MBCs are unavailable can be useful in the effective treatment of serious Enterococcus infections. PMID- 15528892 TI - Effect of ceftazidime, amikacin and ciprofloxacin on biofilm formation by some enterobacterial clinical isolates. AB - The effect of sub-minimal inhibitory concentrations of antimicrobial agents on the biofilm formationto polystyrene by Escherichia coli, Proteus vulgaris, Providencia stuartii, and Morganella morganii was investigated by examining eight clinical strains. All the isolates tested were efficient biofilm-forming strains in the microtiter plate assay, with crystal violet staining (OD595 nm) ranging from 0.13 +/- 0.01 for P. stuartii ER21870 to 1.23 +/- 0.02 for P. vulgaris ER50120. The biofilm formation of the majority of the strains was affected in the presence of ceftazidime or ciprofloxacin: biofilm formation significantly decreased for all the E. coli and P. vulgaris strains in the presence of either of the two antibiotics, it also decreased for M. morganii ER89472 in the presence of ceftazidime but increased for P. stuartii ER21870 and M. morganii ER89472 in the presence of ciprofloxacin. Amikacin decreased only the biofilm formation of P. stuartii ER08274. In addition to their antibacterial activity, ceftazidime and ciprofloxacin could be effective in preventing the biofilm formation of E. coli and P. vulgaris. PMID- 15528894 TI - Peroxisome proliferator-activated receptor-alpha and atherosclerosis: from basic mechanisms to clinical implications. AB - Atherosclerosis is an inflammatory process triggered by the presence of lipids in the vascular wall and encompasses a complex interaction between inflammatory cells, vascular elements and lipoproteins through the expression of several adhesion molecules and cytokines. Activation of the nuclear receptor peroxisome proliferator-activated receptor-alpha (PPAR-alpha) has been demonstrated to modulate many aspects of lipoprotein metabolism and inflammation in vitro as well as in animal and human studies. The tissue distribution of PPAR-alpha is extensive and it is abundantly present in the vascular wall where it may mediate many of anti-inflammatory and antiatherogenic effects. Major clinical trials, such as the Veterans Affairs High-Density Lipoprotein Intervention Trial, the Helsinki Heart Study and the Diabetes Atherosclerosis Intervention Study, have demonstrated the beneficial effects of synthetic agonists of PPAR-alpha, specifically fibric acid derivatives, on cardiovascular disease outcome. Although fibric acid trials have reported cardiovascular risk reduction in patients with dyslipidemia, the favorable alterations in plasma lipids can only partially explain the reduction in cardiovascular events in these studies. One common link among these trials was a cohort with a high prevalence of insulin resistance or diabetes, conditions associated with heightened systemic inflammation and increased risk for development and progression of atherosclerosis. In this paper, we will review the many antiatherogenic effects of PPAR-alpha ligands and evidence from fibric acid trials that individuals with insulin resistance or diabetes benefit the most from these drugs, consistent with their anti inflammatory and antithrombotic properties. PMID- 15528893 TI - Efficacy of quinupristin-dalfopristin against methicillin-resistant Staphylococcus aureus and vancomycin-insensitive S. aureus in a model of hematogenous pulmonary infection. AB - BACKGROUND: Quinupristin-dalfopristin (Q-D) is a mixture of quinupristin and dalfopristin, which are semisynthetic antibiotics of streptogramin groups B and A, respectively. METHODS: We compared the effect of Q-D to that of vancomycin (VCM) in murine models of hematogenous pulmonary infections caused by methicillin resistant Staphylococcus aureus (MRSA) and VCM-insensitive S. aureus (VISA). RESULTS: Treatment with Q-D resulted in a significant decrease in the number of viable bacteria in the lungs of mice in an MRSA infection model [Q-D 100 mg/kg, Q D 10 mg/kg, VCM and control (mean +/- SEM): 2.99 +/- 0.44, 6.38 +/- 0.32, 5.75 +/ 0.43 and 8.40 +/- 0.14 log10 CFU/lung, respectively]. Compared with VCM, high dose Q-D significantly reduced the number of bacteria detected in the VISA hematogenous infection model [Q-D 100 mg/kg, Q-D 10 mg/kg, VCM and control (mean +/- SEM): 5.17 +/- 0.52, 7.03 +/- 0.11, 7.10 +/- 0.49 and 7.18 +/- 0.36 log10 CFU/lung, respectively]. Histopathological examination confirmed the effect of Q D. CONCLUSION: Our results suggest that Q-D is potent and effective in the treatment of MRSA and VISA hematogenous pulmonary infections. PMID- 15528895 TI - Tissue pH determination for the detection of metabolically active, inflamed vulnerable plaques using near-infrared spectroscopy: an in-vitro feasibility study. AB - Detection of vulnerable plaques as the underlying cause of myocardial infarction is at the center of attention in cardiology. We have previously shown that infiltration of inflammatory cells in atherosclerotic plaques renders these plaques relatively hot and acidic, with substantial plaque temperature and pH variation. The objective of this investigation was to determine whether near infrared diffuse reflectance spectroscopy (NIRS) could be used to non destructively measure the tissue pH in atherosclerotic plaques. NIRS and tissue pH electrode measurements were taken on freshly excised carotid plaques maintained under physiological conditions. The coefficient of determination between NIRS and the pH microelectrode measurement was 0.75 using 17 different areas. The estimated accuracy of the NIRS measurement was 0.09 pH units. These results demonstrate the feasibility of using NIRS tissue pH in freshly excised atherosclerotic plaques in light of marked pH heterogeneity and warrants future in-vivo investigations on pH measurement of atherosclerotic plaques. PMID- 15528896 TI - Angiotensin-converting enzyme inhibitor use in elderly patients hospitalized with heart failure and left ventricular systolic dysfunction. AB - PURPOSE: Although angiotensin-converting enzyme (ACE) inhibitors are recommended for all patients with systolic heart failure, prior studies suggest that elderly cohorts are less likely to receive such therapy. The purpose of this study was to determine the age dependence of adherence to guideline-based medical care in hospitalized heart failure patients. METHODS: We performed a multicenter observational cohort study including 613 patients admitted to participating hospitals with a primary diagnosis of heart failure with ejection fraction < or =40%. This cohort was divided into four age groups (group 1: <60, group 2: 60-69, group 3: 70-79, and group 4: 80 years) and adherence to guideline-based medical care was measured. RESULTS: ACE inhibitors were administered to 83% of ideal heart failure patients, and this rate was similar for all age groups. Elderly patients received significantly lower ACE inhibitor dosages compared to their younger counterparts (168, 148, 125 and 117 mg captopril in groups 1, 2, 3, and 4, respectively, p=0.001). Lower creatinine clearance (p<0.001), prior residence in a long-term care facility (p=0.037), intolerance to ACE inhibitors (p=0.006), lower blood pressure (p=0.005), absence of a history of hypertension (p=0.005), and no prior heart failure hospitalizations within the past year (p=0.001) were found to be independent predictors of low ACE inhibitor dosing. CONCLUSIONS: In this heart failure benchmarking project, elderly patients received guideline based ACE inhibitor therapy at similar rates, but at lower doses, compared to their younger counterparts. PMID- 15528897 TI - Implementation of the ACC/AHA guidelines for preoperative cardiac risk assessment in a general medicine preoperative clinic: improving efficiency and preserving outcomes. AB - BACKGROUND: The American College of Cardiology/American Heart Association (ACC/AHA) publishes recommendations for cardiac assessment of patients undergoing noncardiac surgery with the intent of promoting evidence-based, efficient preoperative screening and management. We sought to study the impact of guideline implementation for cardiac risk assessment in a general internal medicine preoperative clinic. METHODS: The study was an observational cohort study of consecutive patients being evaluated in an outpatient preoperative evaluation clinic before and after implementation of the ACC/AHA guideline. Data was gathered by retrospective abstraction of hospital and clinic charts using standard definitions. 299 patients were reviewed prior to guideline implementation and their care compared to 339 consecutive patients after the guideline was implemented in the clinic. RESULTS: Guideline implementation led to a reduction in exercise stress testing (30.8% before, 16.2% after; p<0.001) and hospital length of stay (6.5 days before, 5.6 days after; p=0.055). beta-Blocker therapy increased after the intervention (15.7% before; 34.5% after; p<0.001) and preoperative test appropriateness improved (86% before to 94.1% after; p<0.001). CONCLUSIONS: Implementation of the ACC/AHA guidelines for cardiac risk assessment prior to noncardiac surgery in an internal medicine preoperative assessment clinic led to a more appropriate use of preoperative stress testing and beta blocker therapy while preserving a low rate of cardiac complications. PMID- 15528898 TI - Electrocardiographic pattern as a guide for management and radiofrequency ablation of idiopathic ventricular tachycardia. AB - BACKGROUND: Idiopathic ventricular tachycardia (VT) often originates from the right ventricular outflow tract (RVOT), but foci deep to the endocardium, in the epicardium, or in the left ventricle are not uncommon. Although these extra-RVOT foci can be targeted with ablation, risks involved are higher and success rates lower. Simple electrocardiographic (ECG) criteria allowing (1) discrimination of RVOT foci from extra-RVOT foci and (2) assessment of the chance of success of a right heart ablation procedure are desirable. METHODS: Twenty-five consecutive patients referred for radiofrequency (RF) ablation of idiopathic VT or severely symptomatic idiopathic ventricular premature contractions were included. Localization of VT origin and success rates of VT ablation in the RVOT were analyzed according to the ECG pattern. RESULTS: The analysis of the R wave in V2 was the strongest single predictor of whether the VT had an RVOT or an extra-RVOT origin. An R wave amplitude < or =30% of the QRS amplitude designated the VT focus in the RVOT with positive and negative predictive values of 95 and 100%, respectively. Analysis of R wave duration in V2 had similar predictive values, whereas the R/S transition zone in precordial leads had slightly lower predictive values. Seventeen of 20 arrhythmias (85%) with an R wave amplitude < or =30% of the QRS amplitude in V2 could be successfully abolished by an exclusively right heart procedure. CONCLUSIONS: The analysis of ECG pattern makes it possible to guide the management of patients with idiopathic VT in predicting the arrhythmias that can be safely targeted with RF ablation from the RVOT with high success rates. PMID- 15528899 TI - Resumption and maintenance of employment after a first acute myocardial infarction: sociodemographic, vocational and medical predictors. AB - In a community cohort of 833 women and men aged <60 years, employed at the time of a first acute myocardial infarction (AMI), the relation of sociodemographic, vocational, and medical variables to the following three aspects of employment was investigated longitudinally: resumption of employment anytime within 5 years after AMI, resumption of employment within 3 months of the AMI, and maintenance of employment 5 years after AMI. Different sets of characteristics of persons with a first AMI were related to the different criteria of employment. Only sociodemographic and vocational variables were related to early resumption of employment, whereas maintenance of employment 5 years after AMI was influenced by both medical and nonmedical variables. These results support the importance of examining different aspects of resumption and maintenance of employment after AMI. They also suggest that early resumption of employment after AMI and employment anytime 5 years after AMI are more a function of the sociodemographic and vocational characteristics of the person with the AMI than of that person's medical condition, whereas long-term maintenance of employment after an AMI is also related to the person's medical condition. PMID- 15528900 TI - The outcome of coronary artery bypass grafting surgery among patients hospitalized with acute coronary syndrome: the Euro Heart Survey of acute coronary syndrome experience. AB - AIM: To determine the frequency and outcomes of coronary artery bypass graft (CABG) surgery in patients with a wide spectrum of acute coronary syndromes (ACS). METHODS AND RESULTS: We prospectively enrolled 10,484 ACS patients from 103 hospitals in 25 countries across Europe and the Mediterranean basin. Of the 10,204 patients with complete data, 460 (4.5%) underwent CABG while in hospital; 3.4% had ST elevation ACS, 5.4% had non-ST elevation ACS, and 4.4% had undetermined ECG ACS (p=0.001 for non-ST elevation ACS vs. others). In general, patients who underwent CABG were more likely to be males, to have diabetes mellitus, hyperlipidemia, a positive family history of premature coronary disease, and prior angina pectoris, but had less often prior heart failure. While in hospital, all CABG patients underwent coronary angiography and 15.2% also underwent percutaneous revascularization, as compared with 51.3 and 33.1% in the remaining patients, respectively. The in-hospital mortality was 3.7% for ACS patients who underwent CABG and 4.8% for non-CABG ACS patients (p=nonsignificant) with an adjusted odds ratio of in-hospital death for CABG patients of 1.00 (95% CI 0.59-1.61). CONCLUSIONS: Approximately 4.5% of ACS patients underwent CABG during their initial hospitalization, with a greater likelihood among non-ST elevation ACS patients. Of the CABG patients, 15.2% also underwent percutaneous revascularization. The outcome of CABG patients was as good as non-CABG patients, indicating that CABG remains an effective and safe means to achieve revascularization among ACS patients in current clinical practice. PMID- 15528901 TI - Predictors of cardiovascular death in patients with a left ventricular restrictive filling pattern of the mitral inflow. AB - The survival, clinical and echocardiographic variables and the predictors of cardiovascular death were determined for a group of 168 patients (mean age 63+/ 13 years; 65 females; mean left ventricular ejection fraction 32+/-10%) with restrictive filling of the left ventricle and depressed systolic function after a mean follow-up period of 2.7+/-1 years. Shorter deceleration time (DT) of the mitral inflow was the only variable significantly different between survivors and nonsurvivors (p<0.05) and the only predictor of death found by multivariate logistic regression analysis (odds ratio 2.2, 95% confidence interval 1.7-3.6). In this patient population, a DT of the early wave of the mitral inflow <140 ms identified the patients with the highest risk of cardiac death. DT is a practical echocardiographic parameter for risk stratification of patients with significant left ventricular systolic dysfunction and restrictive filling of the left ventricle. PMID- 15528902 TI - Sudden death due to cardiac tamponade caused by metastasis of squamous cell carcinoma of the lung. PMID- 15528903 TI - Postural orthostatic tachycardia warrants investigation of Chiari I malformation as a possible cause. PMID- 15528904 TI - Diagnosis of pit and fissure caries using frequency-domain infrared photothermal radiometry and modulated laser luminescence. AB - Non-intrusive, non-contacting frequency-domain photothermal radiometry (FD-PTR or PTR) and frequency-domain luminescence (FD-LUM or LUM) have been used with 659- and 830-nm laser sources to assess the pits and fissures on the occlusal surfaces of human teeth. Fifty-two human teeth were examined with simultaneous measurements of PTR and LUM and were compared to conventional diagnostic methods including continuous (dc) luminescence (DIAGNOdent), visual inspection and radiographs. To compare each method to the others, sensitivities and specificities were calculated by using histological observations as the gold standard. With the combined criteria of four PTR and LUM signals (two amplitudes and two phases), it was found that the sensitivity of this method was much higher than any of the other methods used in this study, whereas the specificity was comparable to that of dc luminescence diagnostics. Therefore, PTR and LUM, as a combined technique, has the potential to be a reliable tool to diagnose early pit and fissure caries and could provide detailed information about deep lesions. Using the longer wavelength (830-nm) laser source, it has been shown that detection of deeper subsurface lesions than the 659-nm probe provides is possible. PMID- 15528905 TI - Synchrotron x-ray microtomographic investigation of mineral concentrations at micrometre scale in sound and carious enamel. AB - Synchrotron X-ray microtomography (XMT) was used to measure the linear attenuation coefficient (LAC) for 1.9-microm sidelength voxels within approximal brown spot lesions and sound human enamel. XMT demonstrated three-dimensional features, notably sheets with approximately 30 microm periodicity having low LAC, identified as regions of demineralization corresponding to Retzius lines. Quantitative three-dimensional measurements of mineral concentration, derived from LAC with assumption of a single model composition, were consistent with previous measurements of sound and carious enamel from microradiographic projections. The uncertainty in measurements of mineral concentration and mineral fraction volume was investigated by modelling enamel with a range of composition and component densities. This analysis showed that, although mineral concentration can be determined from LAC with an error of <0.2 g cm(-3), the variation in pore fraction volume within caries lesions cannot be reliably determined from X-ray attenuation measurements alone. PMID- 15528906 TI - Typing of mutans streptococci by arbitrarily primed PCR in patients undergoing orthodontic treatment. AB - The aim of this study was to clarify the genotypic stability of mutans streptococci (MS) longitudinally during orthodontic treatment. Plaque samples were obtained from the supragingival smooth surface of the upper right teeth at four stages: prior to and after 1, 3 and 6 months of orthodontic treatment. Levels of total viable count, total streptococci and MS in dental plaque of 17 patients were recorded. Streptococci isolated from dental plaque samples were identified as MS on the basis of their morphological and biochemical properties. DNA was prepared from 713 strains of MS and the strains were then identified with polymerase chain reaction (PCR) again. Arbitrarily primed PCR (AP-PCR) fingerprinting was applied in determining the genotypes of MS. The results indicated that levels of total viable count, total streptococci and MS increased significantly after the fixed appliances were bonded. A maximum of 3 different genotypes were found in an individual. All the genotypes were found again after the application of the fixed appliances in 17 patients. A new AP-PCR typing pattern was found after the application of fixed appliances for 1 month in patient 1. That strain was not detected either prior to or after 3, or 6 months of treatment. The result indicated that the MS clones were very stable during orthodontic treatment. PMID- 15528907 TI - The photo-activated antibacterial action of toluidine blue O in a collagen matrix and in carious dentine. AB - The main aim of this study was to determine the susceptibility to photo-activated disinfection (PAD) of Streptococcus mutans when the organism was present in a collagen matrix--an environment similar to that which would exist within a carious tooth. In addition, the susceptibility to PAD of bacteria present in carious human teeth was also determined. Light was delivered to the collagen and teeth using a system comprising a 0.8-mm diameter isotropic tip emitting light at 633 +/- 2 nm. A single concentration of TBO (10 microg/ml) was used with both collagen and dentine. Two contact times, 30 and 180 s, were evaluated in intact collagen and additionally, for 180 s only, in collagen partially disrupted by shredding. The effect of energy doses from 1.8 to 14.4 J on the kills attained was assessed by determining the number of surviving viable bacteria. In carious dentine, two contact times, 30 and 60 s and one energy dose, 4.8 J, were used. Antibacterial effects were less than those obtained using planktonic suspensions with a maximum mean log reduction of 1.4 in shredded collagen and dentine. Increasing contact time increased the antibacterial effectiveness in both substrates although this was not always of statistical significance. Shredding the collagen resulted in significantly increased bacterial kills compared to those obtained in intact collagen for the 30-second contact time. The collagen matrix appeared to be a suitable model for carious dentine with advantages of availability and reproducibility. The results of this study have shown that PAD can achieve appreciable kills of oral bacteria, including S. mutans, when the organisms are embedded in a collagen gel or are present in carious teeth. PMID- 15528908 TI - Effect of an essential oil mouthrinse, with and without fluoride, on plaque metabolic acid production and pH after a sucrose challenge. AB - This clinical study evaluated the effect of rinsing with an essential oil containing antiseptic mouthrinse, with or without 100 mg/kg fluoride ion, on the plaque metabolic acid production and plaque pH response after a sucrose challenge. This observer-blind, randomized study used a three-way crossover design. Twenty-four subjects rinsed with 20 ml of one of the following rinses: (1) essential oil (EO) mouthrinse, (2) essential oil mouthrinse plus 100 mg/kg fluoride, or (3) negative control, for 30 s, twice daily for 16 days. On day 17, 1 h after the last mouthrinse, subjects rinsed with 20 ml of mass fraction 10% sucrose solution for 1 min. Seven minutes after the sucrose challenge, supragingival plaque was collected from molar and premolar teeth. Plaque pH and metabolic acid ions were analyzed using a micro pH electrode and capillary electrophoresis, respectively. The results showed that after EO mouthrinse dental plaque produced 36% less lactate, 36% less acetate and 44% less propionate than after the negative control rinse. The dental plaque also exhibited a pH 0.42 unit higher after EO rinse than after the negative control rinse. These results were not affected by the addition of 100 mg/kg fluoride to the EO mouthrinse. From these results we concluded that this EO antiseptic mouthrinse, with or without fluoride ion, is effective in reduction of plaque acidogenicity after a sucrose challenge. PMID- 15528909 TI - The relative anticaries effectiveness of three fluoride-containing dentifrices in Puerto Rico. AB - The objective of this research was to evaluate the anticaries effectiveness of a low-dose (500 ppm F, low-NaF) sodium fluoride dentifrice, a high-dose (2,800 ppm F, high-NaF) sodium fluoride dentifrice and an experimental 0.454% stabilized stannous fluoride (1,100 ppm F) with sodium hexametaphosphate (SnF2-HMP) dentifrice, each relative to a standard 1,100 ppm F sodium fluoride positive control dentifrice. Subjects (n = 955, with approximately 239 per group) with a mean age of 10.6 (approximately 9-12 years) were randomly assigned to one of four dentifrice treatments. Two calibrated examiners independently measured visual tactile caries as DMFS that was supplemented with a radiographic examination at baseline, 12 months and 24 months for each subject. Generally similar results were independently observed by both examiners at the conclusion of the 2-year study period. Considering all subjects that attended at least 60% of the supervised brushing sessions, statistically significantly less caries was observed in the high-NaF group compared to the control group. Similarly, statistically significantly less caries was observed in the SnF2-HMP group as compared to the control group. Differences in caries increments between the low NaF and control groups were not statistically significant. One of the examiners observed these same statistically significant differences after 1 year. In conclusion, the results of this clinical trial indicated that while no difference in caries increments was observed between the low-NaF and control groups, both the high-NaF and the SnF2-HMP groups experienced significantly fewer lesions than the control group. PMID- 15528910 TI - Acid resistance of enamel subsurface lesions remineralized by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate. AB - The aim of this clinical study was to investigate the acid resistance of enamel lesions remineralized in situ by a sugar-free chewing gum containing casein phosphopeptide-amorphous calcium phosphate nanocomplexes (CPP-ACP: Recaldent). The study utilized a double-blind, randomized, crossover design with two treatments: (i) sugar-free gum containing 18.8 mg of CPP-ACP, and (ii) sugar-free gum not containing CPP-ACP as control. Subjects wore removable palatal appliances with insets of human enamel containing demineralized subsurface lesions and chewed the gum for 20 min 4 times per day for 14 days. After each treatment the enamel slabs were removed and half of each lesion challenged with acid in vitro for 8 or 16 h. The level of remineralization was determined using microradiography. The gum containing CPP-ACP produced approximately twice the level of remineralization as the control sugar-free gum. The 8- and 16-hour acid challenge of the lesions remineralized with the control gum resulted in 65.4 and 88.0% reductions, respectively, of deposited mineral, while for the CPP-ACP remineralized lesions the corresponding reductions were 30.5 and 41.8%. The acid challenge after in situ remineralization for both control and CPP-ACP-treated lesions resulted in demineralization underneath the remineralized zone, indicating that the remineralized mineral was more resistant to subsequent acid challenge. The results show that sugar-free gum containing CPP-ACP is superior to an equivalent gum not containing CPP-ACP in remineralization of enamel subsurface lesions in situ with mineral that is more resistant to subsequent acid challenge. PMID- 15528911 TI - A study in vitro of the abrasive effect of the tongue on enamel and dentine softened by acid erosion. AB - The palatal aspect of upper anterior teeth appears to be one site of predilection for erosion. This raises the question of whether abrasion of softened enamel and dentine by the tongue has a role in this process. The aim of this study in vitro was to determine whether enamel and dentine specimens licked by the tongue after exposure to acid lost more tissue than when exposed to acid alone. Flat enamel and dentine specimens were prepared from human third molars and taped to expose a window of tissue. Three groups of specimens were exposed to citric acid for 10 min followed by 60 s tongue licking, ultrasonication or immersion in water. Two further groups of enamel specimens were placed in a low erosive drink for 10 min with or without licking. At the end of 5, 10 and 15 treatment cycles tissue loss was measured by profilometry. Enamel loss was significantly greater with licking and ultrasonication compared to water immersion. Dentine loss was greater with licking and ultrasonication compared to water immersion, but differences only reached significance for ultrasonication. The low erosive drink produced one third of the citric acid erosion and licking had no effect. The results suggest that the tongue could exert an abrasive effect on dental tissues softened by erosion, thereby increasing the overall loss of tooth substance. PMID- 15528913 TI - Effect of amine/sodium fluoride rinsing on toothbrush abrasion of softened enamel in situ. AB - The aim of this study was to test the effect of fluoride rinsing on the prevention of toothbrush abrasion of softened enamel in situ. For softening, the samples were immersed in 0.1 M citric acid (pH 3.5) for 3 min. Eight test subjects had to make 5 runs in which 4 slabs per run were attached to intraoral appliances. They were as follows: (1) no softening, no fluoride rinsing (control 1); (2) softening, no fluoride rinsing (control 2); (3) softening, rinsing in situ with a sodium/amine fluoride rinsing solution (250 ppm F) for 30 s; (4) rinsing in situ with the sodium/amine fluoride rinsing solution (250 ppm F) for 30 s, softening; (5) softening, rinsing in situ with an experimental amine fluoride-containing rinsing solution (250 ppm F) for 30 s. After exposure for 60 min to the oral milieu, the volunteers brushed the samples for 30 s with toothpaste and the loss of tooth substance was determined. For each test person, the secretion rate of resting and paraffin-stimulated saliva, buffering capacity and pH were measured. Toothbrush abrasion in situ was not significantly lower using the fluoride rinsing solutions before or after softening the enamel compared to no rinsing (p > 0.05). Multiple linear regression analyses revealed that 57% of the variation in toothbrush abrasion could be attributed to the severity of softening (p < 0.001) and the pH of stimulated saliva (p < 0.001). It was concluded that a single rinse for 30 s had no statistically significant effect on the prevention of toothbrush abrasion of softened enamel. PMID- 15528912 TI - Effects of two fluoridation measures on erosion progression in human enamel and dentine in situ. AB - The aim of the present study was to evaluate the effects of fluoride on erosive mineral loss in human enamel and dentine using a cyclic de- and remineralisation model in situ. The study was a three-treatment (5 days each) crossover design involving 4 (enamel) or 6 (dentine) healthy volunteers. Samples were recessed in palatal mouth appliances and worn day and night except during meals and were demineralised extraorally with 0.05 M citric acid (pH 2.3) for 6 x 5 min daily. Fluoridation was performed with toothpaste (SnF2/Olaflur; 0.14% F-) for 3 x 5 min daily (toothpaste fluoridation) or with toothpaste in combination with a mouthrinse (SnF2/Olaflur; 0.025% F-) for 3 x 5 min daily and with a gel (NaF/Olaflur, 1.25% F-) on days 1 and 3 instead of the toothpaste (intensive fluoridation). In the control group no fluoridation was performed. Mineral loss (microm) was determined with the use of longitudinal microradiography. In enamel, mineral loss was 40.7 +/- 15.1 microm in the control group, 18.3 +/- 12.4 microm after toothpaste fluoridation and 5.0 +/- 12.2 microm after intensive fluoridation. The respective values for dentine were 49.0 +/- 15.4, 35.0 +/- 15.5 and 19.8 +/- 12.0 microm. All differences were statistically significant (p < or = 0.001). The results indicate that intensive fluoridation is effective in preventing enamel and dentine from mineral loss even under severely erosive conditions. PMID- 15528915 TI - No relationship between subchronic fluoride intake and DNA damage in Wistar rats. AB - Fluoride has been widely used in dentistry because it is an effective caries prophylactic agent. However, excess fluoride may represent a hazard to human health, especially by causing injury on the genetic apparatus. Genotoxicity tests form an important part of cancer research and risk assessment of potential carcinogens. In the current study, the potential DNA damage associated with exposure to fluoride was assessed by the single cell gel (comet) assay in peripheral blood, oral mucosa and brain cells in vivo. Male Wistar rats were exposed to sodium fluoride (NaF) at a 0, 7 and 100 ppm dose for drinking water during 6 weeks. The results pointed out that NaF did not contribute to the DNA damage in all cellular types evaluated as depicted by the mean tail moment and tail intensity. These findings are clinically important since they represent an important contribution to the correct evaluation of the potential health risk associated with dental agents exposure. PMID- 15528914 TI - Caries prevalence, levels of mutans streptococci, and gingival and plaque indices in 3.0- to 5.0-year-old mouth breathing children. PMID- 15528916 TI - Relationship between retinopathy and cognitive impairment among hypertensive subjects. A case-control study in the ankara-pursaklar region. AB - BACKGROUND: The relationship between hypertension and cognitive impairment has been investigated in the literature; several clinical studies suggested a relationship between hypertension and retinopathy. METHODS: We examined the relationship between the retinopathy status and cognitive functions by using the Mini-Mental State Examination (MMSE) among hypertensive subjects older than 40 years who were admitted to our Family Medicine, Internal Medicine and Ophthalmology clinics. The subjects were categorized into three groups: group 1 = control subjects (n = 39), group 2 = hypertensive patients without retinopathy (n = 32), and group 3 = hypertensive patients with retinopathy (n = 25). RESULTS: The number of patients with total MMSE scores less than 24 was distributed as follows: group 1: 3 patients (7.7%), group 2: 4 patients (12.5%), and group 3: 14 patients (56%). Hypertension was found to be related with a significant decrease in total MMSE scores in comparison with group 1 subjects (p < 0.001). Furthermore, retinopathy significantly correlated with lower MMSE scores among hypertensive patients (p = 0.001). Compared with group 1, group 3 showed a significant decrease in orientation (p = 0.001), registration (p = 0.001), attention and calculation (p = 0.004), recall (p = 0.009), and total (p < 0.001) MMSE scores. We also found a significant decrease in recall (p = 0.032) and total (p = 0.034) scores in group 3 when compared with group 2. There was a significant decrease in registration (p = 0.002) and total (p = 0.029) MMSE scores in group 2 when compared with group 1. We also observed negative correlations between duration of the disease and orientation (R = -0.597, n = 32, p = 0.001), and between duration of the disease and total (R = -0.495, n = 32, p = 0.006) scores in group 2. CONCLUSIONS: Hypertension was found to be related with a decline in MMSE scores. This relation was even more significant in the group of hypertensive patients with retinopathy. Thus, we suggest that cognitive tests be routinely used in the follow-up of hypertensive patients. PMID- 15528917 TI - Expression of TH1/TH2-related chemokine receptors on peripheral T cells and correlation with clinical disease activity in patients with multiple sclerosis. AB - Th1 cells play an important role in the pathogenesis of multiple sclerosis (MS), a disease likely linked to an autoimmune process. We measured the levels of chemokines in serum or cerebrospinal fluid (CSF) samples by ELISA, and also studied the expression of Th1-related CXCR3/CCR5 chemokine receptors and Th2 related CCR4/CCR3 chemokine receptors on blood cells from MS patients using three color flow cytometry. The Bonferroni correction was used for the statistical analysis. The levels of CXCL10, CCL3, and CCL5 in the CSF samples for the MS groups were significantly higher than those for the control group. However, the levels of CCL2 in both the CSF and serum samples for the remission group were significantly higher than those for the active group. The percentage of CXCR3 expressing CD4+ T cells in patients with MS was significantly elevated compared with the healthy controls. Moreover, MS patients in an active phase showed a more increased CD4+CXCR3+/CD4+CCR4+ ratio than patients in a remission phase. The increased percentage of CD4+CXCR3+ cells in the blood was associated with relapses in MS. This study suggested that the CD4+CXCR3+/CD4+CCR4+ ratio could be a sensitive maker of immune dysfunction in MS. PMID- 15528918 TI - Some neurological aspects of laughter. AB - This brief survey of laughter attempts an analysis of its neurological mechanisms, evolution, role in social behaviour and its clinicopathological importance. The mechanisms of laughter, its physiological consequences and its demonstration by sound spectrography are considered. Something resembling laughter occurs in certain primates, and possibly rodents, though there are important differences. The evolution of laughter in a social context is appraised. Pathological laughter arises rarely, usually caused by diseases of the frontal or temporal lobes, and in hypothalamic hamartomata in children. PMID- 15528919 TI - A note on gelastic epilepsy. AB - Laughter epilepsy or gelastic seizures have been described in various epilepsies arising from the temporal or frontal lobes, but most commonly from hypothalamic hamartomata. Gelastic seizures also arise from temporal and frontal lobe tumours and atrophic lesions. The essential clinical features are: stereotyped recurrence; absence of external precipitants; concomitance of other manifestations generally accepted as epileptic; presence of interictal or interictal EEG epileptiform discharges, and absence of conditions in which pathological laughter might occur. The history and clinical significance are discussed. PMID- 15528920 TI - Phenotype variability in a caucasian family with dentatorubral-pallidoluysian atrophy. PMID- 15528921 TI - A sensitive way to demonstrate subtle optic ataxia. PMID- 15528922 TI - Valproate-induced hyperammonemic encephalopathy: imaging findings on diffusion weighted MRI. PMID- 15528923 TI - Brainstem infarction during long-haul air travel. PMID- 15528924 TI - Bilateral axonal facial nerve lesion after sagittal split osteotomy: a case report. PMID- 15528925 TI - Acute transverse myelitis as a main manifestation of early stage II neuroborreliosis in two patients. PMID- 15528926 TI - Endocrine and cognitive effects of short-time soybean consumption in women. AB - BACKGROUND: Soy phytoestrogens are known to influence the hormonal status acting as partial estrogen agonists. Soy-derived food supplements are advised for hormone replacement therapy, prevention of atherosclerosis, age-related cognitive decline and even hormone-dependent cancer, although results from clinical studies are controversial. Whether increased soybean intake can affect the endocrine status and cognitive abilities is largely unknown. AIM: To observe the effects of 1 week of increased soybean intake on sex hormone levels and spatial cognitive abilities in women. SUBJECTS AND METHODS: 16 young healthy female volunteers were asked to eat 900 g of soybeans within 1 week. Salivary testosterone (T), free and total plasma T, salivary and plasma estradiol (E) were measured by radioimmunoassay before and after the study period. Mental rotation (MR) and spatial visualization (SV) psychological tests were done at the days of sampling. RESULTS: Soybean intake increased total plasma T levels (p < 0.02) while decreasing salivary T (p < 0.01) and not altering free plasma T levels. Salivary and plasma E levels were not changed. The results of MR and SV tests were improved after the study period. CONCLUSION: Short-time increased soybean intake alters the level of total plasma and salivary T and improves spatial cognition in women. Whether this effect is mediated by modulation of estrogen receptors, changes in sex hormone-binding globulin production or changes in activity of steroid-competent enzymes needs further study. PMID- 15528927 TI - Clinical, biochemical and morphologic diagnostic markers in an infant male pseudohermaphrodite patient with compound heterozygous mutations (G115D/R246W) in SRD5A2 gene. AB - A patient with male pseudohermaphroditism and clinical diagnosis of partial androgen insensitivity in the neonatal period was studied at pubertal age for a molecular diagnosis. Hormone studies were conducted at baseline and under hCG stimulation for testosterone and dihydrotestosterone determinations at 2 months of age. Gonadectomy was performed at 4 months. At the age of 13 years genital skin fibroblasts were studied for androgen binding and 5alpha-reductase activity and peripheral blood DNA was available for androgen receptor (AR) and 5alpha reductase (SRD5A2) gene analysis. Exons 1-8 of AR gene and exons 1-5 of SRD5A2 gene were sequenced. AR gene coding sequences were normal. SRD5A2 gene analysis revealed two heterozygote mutations (G115D and R246W), with the mother carrying the G115D and the father the R246W mutations. The compound heterozygote mutations in SRD5A2 gene explained an extremely low 5alpha-reductase enzyme activity in genital skin fibroblasts. Revision of hormonal data from the neonatal period revealed an increased testosterone-to-dihydrotestosterone ratio at the end of an hCG stimulation test, which concurred with the molecular diagnosis. Testis morphology at 4 months of age was normal. Clinical and biochemical differential diagnosis between partial androgen insensitivity syndrome and 5alpha-reductase enzyme deficiency is difficult in the neonatal period and before puberty. Our results show that in our patient the testosterone-to-dihydrotestosterone ratio would have adequately orientated the diagnosis. The two mutations in SRD5A2 gene have been described in patients of different lineages, though not in combination to date. Testis morphology showed that, during early infancy, the 5alpha reductase deficiency may not have affected interstitial or tubular development. PMID- 15528928 TI - Allergenic potency of spices: hot, medium hot, or very hot. AB - Spices are the most attractive ingredients to confer an authentic taste to food. As they are derived from plants, they harbour allergenic potency and can induce symptoms ranging from mild local to severe systemic reactions. Due to the content of pharmacologically active substances of spices, the diagnosis of allergy and the differentiation from intolerance reactions may be difficult. Association with inhalative allergies via IgE cross-reactivity, but also direct gastrointestinal sensitization plays a role. This article is a botanical and allergological overview of the most important spices and molecules responsible for eliciting IgE mediated reactions or cross-reactions. As no curative treatments are known at present, strict avoidance is recommended and, therefore, accurate labelling of pre-packed food is necessary. PMID- 15528930 TI - Plasminogen activator expression correlates with genetic differences in vascular remodeling. AB - Intima-media thickening (IMT) of the carotid artery, a form of vascular remodeling, correlates well with coronary artery disease risk in humans. Vascular remodeling in response to blood flow is a complex process that critically involves altered cell matrix interactions. To gain insight into these events, we performed partial carotid ligation (left carotid (LCA) = low flow and right carotid (RCA) = high flow) in 2 inbred mouse strains: C57Bl/6J (C57) and FVB/NJ (FVB). To evaluate the role of the 2 major matrix-degrading systems, plasminogen activators (PAs) and matrix metalloproteinases (MMPs), we compared the expression of u-PA, t-PA, MMP-2 and MMP-9 in ligated carotids of C57 and FVB mice. The extent of remodeling was greater in response to low LCA than high RCA flow. Despite a similar decrease in LCA flow in both strains, maximal IMT volume was greater in FVB (82 +/- 7 x 10(-6) microm(3)) than in C57 (38 +/- 4 x 10(-6) microm(3)) after ligation. Among PAs and MMPs, increased expression of t-PA and u PA correlated with increased IMT (p < 0.0005 and p < 0.001, respectively). MMP-2, MMP-9 and tissue inhibitors of metalloproteinase-2 expression also increased, but did not differ between strains. In summary, flow-induced IMT of the carotid is genetically determined and correlates with t-PA and u-PA expression in 2 inbred mouse strains. PMID- 15528931 TI - TGF-beta1, -beta2 and -beta3 cooperate to facilitate tubulogenesis in the explanted quail heart. AB - BACKGROUND: Transforming growth factor-beta (TGF-beta) isoforms have been implicated as both pro- and anti-angiogenic modulators. In this study we addressed the roles of TGF-beta isoforms on coronary tubulogenesis. METHODS: Embryonic (E6) quail ventricular specimens were explanted onto collagen gels allowing endothelial cells to migrate and form vascular tubes. Growth factors and/or neutralizing growth factor antibodies were added to the cultures. Endothelial cells were identified using a quail endothelial cell marker, QH1. Image analysis was used to quantify aggregate tube length. RESULTS: Addition of any isoform (TGF-beta(1), TGF-beta(2) or TGF-beta(3)) virtually prevented tubulogenesis (>95% inhibition), while stimulation of tubulogenesis occurred by adding neutralizing antibodies to TGF-beta(3), but not to TGF-beta(1) or beta(2). When all three isoforms were added, tubulogenesis was enhanced, indicating the key role of TGF-beta(3). Documentation of the inhibitory effect of TGF-beta isoforms on tubulogenesis is further supported by our experiments in which the marked enhancement of tube formation by bFGF and VEGF was negated when exogenous TGF-beta(1), -beta(2), or -beta(3) were added to the cultures. CONCLUSIONS: (1) TGF-beta(1), -beta(2) and -beta(3) each inhibits angiogenesis; (2) cooperation between the three TGF-beta isoforms and other angiogenic factors is essential for the regulation of normal tubulogenesis and (3) the stimulatory effect of VEGF or bFGF on tubulogenesis is negated by exogenous TGB-betas. PMID- 15528932 TI - Decorin deficiency leads to impaired angiogenesis in injured mouse cornea. AB - Small leucine-rich proteoglycans play important roles in the organization of the extracellular matrix as well as for the regulation of cell behavior; two biological processes that are essential for angiogenesis. We investigated consequences of the targeted ablation of decorin (DCN), biglycan (BGN) and fibromodulin (FMOD) genes on inflammation-induced angiogenesis in the cornea. In wild-type mice, DCN was localized exclusively to the corneal stroma, while FMOD and BGN were more prominently expressed in epithelial cells. Endothelial cells from limbus blood vessels expressed BGN and FMOD, but no DCN. However, after induction of angiogenesis by chemical cauterization, DCN was expressed in the newly formed capillaries, together with BGN and FMOD. Notably, in DCN-deficient mice, the growth of vessels was significantly diminished, whereas it did not significantly change in FMOD- or BGN-deficient animals. Moreover, blood vessels of DCN-deficient mice exhibited a similar expression level of BGN as control mice, while FMOD was increased on day 3 after injury. These results indicate that DCN, in addition to its effects on fibrillogenesis, plays a regulatory role in angiogenesis and that FMOD in endothelial cells may be able to partially substitute for DCN. PMID- 15528933 TI - Effects of the dietary flavonoid chrysin in isolated rat mesenteric vascular bed. AB - In the present study, the effects of the bioflavonoid chrysin (5,7 dihydroxyflavone) were analyzed on the perfusion pressure of isolated mesenteric vascular bed. The vasorelaxant effects of chrysin were more potent on intact endothelium than on denuded vessels. This endothelium-dependent response induced by chrysin was inhibited in the presence of N(G)-nitro-L-arginine methyl ester (L NAME), KCl, tetraethylammonium (TEA), BaCl(2), TEA plus L-NAME, and ouabain plus BaCl(2), while incubations with indomethacin and glibenclamide did not modify the response induced by this bioflavonoid. Neither gap junction inhibition with carbenoxolone nor epoxyeicosatrieconic acid synthesis inhibition with sulfaphenazole (selective CYP 2C/3A inhibitor) or 7-ethoxyresorufin (selective CYP 1A inhibitor) inhibited the chrysin-induced relaxation. Moreover, chrysin increased L-NAME-sensitive cGMP accumulation in intact vascular mesenteric preparation. In conclusion, chrysin shows vasodilator effects on resistance vessels, which depend partially on the functional endothelium and appear to be related to the NO/cGMP pathway and, possibly to the release of endothelium derived hyperpolarizing factor. PMID- 15528934 TI - The conduction of dilation along an arteriole is diminished in the cremaster muscle of hypertensive hamsters. AB - Arteriolar vasomotor responses can include a component that conducts along the vessel through gap junction channels. This study examined conducted vasomotor responses in arterioles of the hypertensive hamster. The cremaster muscle of normotensive (CHF-148) and spontaneously hypertensive (CHF-H4) hamsters was exteriorized. Micropipettes containing phenylephrine (0.1 M) or acetylcholine (ACh; 1.0 M) were positioned along second-order arterioles and diameter responses were recorded locally for every 0.4 mm upstream to 1.6 mm. Substantative local constrictions to phenylephrine(PE) were poorly conducted to the 0.4-mm site in normotensive and hypertensive hamsters. Local dilation to ACh decayed by 3 +/- 1 microm/mm as it conducted along arterioles of the normotensive hamster. In contrast, conducted dilation decayed by 7 +/- 1 microm/mm (p < 0.05) in the hypertensive hamster. This hypertension-induced increase in decay was reversed by alpha-adrenergic receptor blockade (phentolamine: 1 microM). However, arteriolar constriction to global alpha(1)- (PE) and alpha(2)- (clonidine) adrenergic agonists was unaffected by hypertension. Rather, sympathetic nervous activity was elevated in the hypertensive hamster as indicated by a greater reduction in arterial pressure upon sympathetic ablation (hexamethonium infusion: 30 mg/kg). This study provides the first evidence that vascular cell-cell communication is altered by the elevated sympathetic nervous activity observed in the hypertensive hamster. PMID- 15528935 TI - Pathogen burden, inflammation, proliferation and apoptosis in human in-stent restenosis. Tissue characteristics compared to primary atherosclerosis. AB - Pathogenic events leading to in-stent restenosis (ISR) are still incompletely understood. Among others, inflammation, immune reactions, deregulated cell death and growth have been suggested. Therefore, atherectomy probes from 21 patients with symptomatic ISR were analyzed by immunohistochemistry for pathogen burden and compared to primary target lesions from 20 stable angina patients. While cytomegalovirus, herpes simplex virus, Epstein-Barr virus and Helicobacter pylori were not found in ISR, acute and/or persistent chlamydial infection were present in 6/21 of these lesions (29%). Expression of human heat shock protein 60 was found in 8/21 of probes (38%). Indicated by distinct signals of CD68, CD40 and CRP, inflammation was present in 5/21 (24%), 3/21 (14%) and 2/21 (10%) of ISR cases. Cell density of ISR was significantly higher than that of primary lesions (977 +/- 315 vs. 431 +/- 148 cells/mm(2); p < 0.001). There was no replicating cell as shown by Ki67 or PCNA. TUNEL(+) cells indicating apoptosis were seen in 6/21 of ISR specimens (29%). Quantitative analysis revealed lower expression levels for each intimal determinant in ISR compared to primary atheroma (all p < 0.05). In summary, human ISR at the time of clinical presentation is characterized by low frequency of pathogen burden and inflammation, but pronounced hypercellularity, low apoptosis and absence of proliferation. PMID- 15528936 TI - Skin blood flow in the upper and lower extremities of diabetic patients with and without autonomic neuropathy. AB - BACKGROUND: Microvascular blood flow in the human skin is subject to rhythmic variations reflecting the influence of heartbeat, respiration, intrinsic myogenic activity, neurogenic factors and endothelial activity. The aim of our study was to test the hypothesis that basal skin blood flow (BSBF) and its dynamic components differ (1) among diabetic patients without autonomic neuropathy and with it and healthy control subjects, and (2) among the upper and lower extremities. PATIENTS AND METHODS: BSBF at four recording sites with predominantly nutritive capillary circulation (right and left caput ulnae, right and left medial malleolus) was measured by laser Doppler flowmetry in 25 diabetic patients without cardiovascular autonomic neuropathy (D), 18 neuropathic diabetic patients (DAN) and 36 healthy controls (C). Wavelet transform was applied to the laser Doppler signal. RESULTS: In absolute terms, mean flow, mean amplitude of the total spectrum and mean amplitudes at all frequency intervals were highest in C, followed by DAN and lowest in D. However, these differences were statistically significant only in the left arm. Within all three groups, mean flow and spectral amplitudes were significantly higher in the arms than in the legs, besides there was a significant difference between the two arms in D. CONCLUSION: We have confirmed the differences in BSBF among D, DAN and C, and demonstrated differences among the four recording sites which have not been previously described. The latter indicates an uneven progression of autonomic neuropathy and allows for speculation that the left arm is the latest to be affected. PMID- 15528937 TI - Mycophenolate mofetil and post-transplant glomerular disease. PMID- 15528938 TI - High-dose mycophenolate mofetil in the treatment of posttransplant glomerular disease in the allograft: a case series. AB - BACKGROUND: Glomerular disease is an important cause of allograft loss. Treatment regimens for posttransplant glomerular disease are not well defined. Several reports have demonstrated that mycophenolate mofetil (MMF) is effective in treating native kidney glomerular disease. The effects of MMF are dose related. Therefore, we hypothesized that high-dose MMF (3 g/day) would be effective in treating glomerular disease in the allograft, minimizing the need for intravenous steroids and/or cyclophosphamide. This case series describes the results of the use of high-dose MMF in 6 patients. METHODS: High-dose MMF (3 g/day) was used to treat biopsy-proven glomerular disease (focal segmental glomerulosclerosis, membranoproliferative glomerulonephritis, proliferative lupus nephritis, and perinuclear antineutrophil cytoplasmic antibodies glomerulonephritis) in 6 renal transplant recipients. Patients were offered this treatment if they had failed or did not tolerate standard treatment regimens. Remission was defined by a decrease or stabilization of serum creatinine, decrease in proteinuria and, where applicable, improvement in immunological markers of disease. RESULTS: All 6 patients had disease remission after starting MMF with the most common side effect being leukopenia, which responded to dose reduction. CONCLUSIONS: High dose MMF may be an effective agent in treating glomerular disease in the allograft. PMID- 15528939 TI - Color Doppler ultrasound in renal transplant: role of resistive index versus renal cortical ratio in the evaluation of renal transplant diseases. AB - BACKGROUND/AIMS: Ultrasound (US) and color Doppler are not sensitive enough to detect anomalies in cortical perfusion, which is affected in most graft dysfunctions. The renal cortical ratio (RCR) is a variation in the resistive index (RI) values from the renal artery to cortical vessels, expressed in percent. The aim of this study was to compare the RI and RCR in the differentiation of normal and pathological grafts, to assess the positive predictive value of RCR and show that RCR enables earlier diagnosis than RI. METHODS: Based on clinical, biochemical and histological examinations, 494 renal allografts were divided into 3 groups (normal grafts, acute and chronic pathologies). All patients underwent US color Doppler. RI was measured and RCR calculated. Follow-up confirmed the initial division in groups. Statistical significance was calculated using the two-tailed Student's t test. The positive predictive value was calculated for each group. RESULTS: 24 h after transplant, RCR differentiated normal grafts from acute dysfunctions despite confusing biochemical values and clinical symptoms. In chronic patients, RCR variations occurred later but always before the serum creatinine level increased. CONCLUSION: RCR presented a higher positive predictive value than RI. RCR curves were already altered in the early stages of transplant pathologies. RCR calculation is easy and makes a significant contribution towards a correct early diagnosis. PMID- 15528940 TI - Urinary excretion of vascular endothelial growth factor is increased in children with reflux nephropathy. AB - BACKGROUND: We determined the urinary levels of vascular endothelial growth factor (VEGF) in patients with reflux nephropathy to elucidate its clinical significance as a predictor of the development of renal damage. PATIENTS AND METHODS: Seventy-two patients (47 boys and 25 girls) aged 10-18 years with reflux nephropathy were studied. Vesicoureteral reflux was resolved surgically in all cases at least 2 years before enrollment. Urinary VEGF, alpha1-microglobulin and albumin levels were determined using morning spot urine samples. Plasma VEGF, serum creatinine and beta2-microglobulin levels were measured simultaneously. The severity of renal scarring and right and left kidney function were assessed by 99mTc dimercaptosuccinic acid renal scan. RESULTS: No significant correlation was found between the plasma and urinary VEGF levels. Urinary VEGF increased significantly with the increase in severity of renal scarring (p < 0.0001). Urinary VEGF levels correlated significantly with serum beta2-microglobulin (p < 0.002) and urinary alpha1-microglobulin (p < 0.03). No significant correlation of urinary VEGF levels with serum creatinine and urinary albumin levels was found. Nearly 60% of the patients with elevated urinary VEGF had normal levels of serum beta2-microglobulin and/or urinary alpha1-miocroglobulin. CONCLUSIONS: Urinary VEGF level appears to reflect its production in the kidney. Since urinary VEGF shows a propensity to elevate before the increase in serum beta2-microglobulin and/or urinary alpha1-microglobulin, urinary VEGF may serve as an early indicator of the development of reflux nephropathy. PMID- 15528941 TI - Could plasma cystatin C be useful as a marker of hemodialysis low molecular weight proteins removal? AB - BACKGROUND: Plasma cystatin (pCyst) is a well-assessed tool for measuring renal function, and it could also play a part in hemodialysis adequacy. METHODS: pCyst and other uremic toxins (urea, creatinine, parathyroid hormone, prolactin) were assessed before and after a dialysis session in 18 hemodialysis patients: 7 on bicarbonate hemodialysis (BHD) and 11 on mixed convective dialysis (MCD; 6 standard hemodiafiltration and 5 acetate-free biofiltration). Plasma levels and reduction ratios (RR) were then compared between the BHD and MCD groups. RESULTS: The mean pre-dialysis pCyst level is nearly the same in both groups (5.3 +/- 0.8 vs. 5.7 +/- 1 mg/l, p = ns), although a substantial decrease occurs after MCD only (mean 2.4 +/- 1 vs. 6.2 +/- 2.2 mg/l after BHD, p = 0.002). The mean pCyst RR (PCRR) of 55.5% after MCD is poorly related to prolactin and urea RR, fairly comparable to parathyroid hormone RR and very close to creatinine RR (58.4%). CONCLUSIONS: Only MCD removes pCyst, but the amount of removal is different for other low molecular weight proteins (prolactin and parathyroid hormone) and similar for creatinine, a classic 'little molecule'. In view of the discrepancy of these findings, the use of pCyst in hemodialysis still seems premature and needs further studies. PMID- 15528942 TI - Effects of intravenously administered iron on systemic blood pressure in hemodialysis patients. AB - BACKGROUND/AIMS: Although maintenance of iron stores by intravenous iron infusion has become increasingly common in chronic hemodialysis patients, the effects of intravenous iron on blood pressure are not clear. METHODS: In 52 mostly African American patients in an urban setting, we reviewed blood pressure data over the course of hemodialysis on days that intravenous iron was administered and on control days. RESULTS: The drop in blood pressure from the beginning to the end of hemodialysis was more pronounced when iron was not administered (9.9 +/- 1.2 mm Hg) compared to when iron was administered (4.1 +/- 1.7 mm Hg; p < 0.01). In a subset of chronic hemodialysis patients, blood pressure was actually higher at the end of hemodialysis compared to at the onset (9.6% of control hemodialysis sessions and 32.7% of sessions in which iron was administered; p < 0.01). The incidence of intra-dialytic hypotension and related symptoms was similar on iron and non-iron days. CONCLUSION: Intravenous iron therapy, well entrenched in clinical practice for the treatment of the anemia of end-stage renal disease, may reduce the incidence and magnitude of post-dialytic hypotension. PMID- 15528943 TI - Significance of acute versus chronic troponin T elevation in dialysis patients. AB - INTRODUCTION: Cardiac troponin T (cTnT) is often elevated in hemodialysis (HD) patients without acute coronary syndrome (ACS). The aim was to assess the predictive value for mortality of pre-dialysis cTnT in asymptomatic patients. If patients became symptomatic during follow-up, cTnT was followed to assess its diagnostic value for ACS. METHODS: Forty-nine asymptomatic HD patients were included: 30 patients with a history of cardiovascular disease (CV+) and 19 without (CV-). In 11 patients cTnT, myoglobin and creatine kinase (CK) were measured before and during HD. During ACS, cTnT was followed until recovery. A cTnT of > or =0.03 mug/l was considered elevated. Follow-up was 2 years. RESULTS: cTnT was elevated in 82% (40/49). More CV+ patients had an elevated cTnT (28/30) than CV- patients (12/19; p = 0.02). There was no change in cTnT, myoglobin and CK during HD. During ACS, cTnT increased above baseline, and tended to return to baseline after recovery. Mortality was 33% (16/49). Patients with elevated cTnT had a higher mortality rate (16/40) than patients with negative cTnT (0/9; p = 0.02). CONCLUSIONS: Elevated cTnT levels in asymptomatic HD patients are not caused by acute myocardial injury or by HD itself. They may be related to chronic myocardial damage and decreased clearance, and are of prognostic value. During ACS, however, a cTnT rise above the individual baseline is diagnostic of acute myocardial injury. PMID- 15528944 TI - Intravenous vitamin D therapy reduces PTH-(1-84)/large C fragments ratio in chronic hemodialysis patients. AB - BACKGROUND: Renal osteodystrophy is one of the major complications in patients with chronic renal failure. Large C-PTH fragments are secreted from the parathyroid glands and exert antagonistic actions against PTH-(1-84). The PTH-(1 84)/large C-PTH fragments ratio reflects both biosynthesis and processing of PTH; however the alteration of the ratio under vitamin D therapy has not been investigated. METHODS: Seventeen hemodialysis patients with intact PTH levels of >300 pg/ml were enrolled. Calcitriol or maxacalcitol were administered intravenously for 78 weeks. Intact PTH, PTH-(1-84), and the PTH-(1-84)/large C PTH fragments ratio were measured at 0, 13, 26, 52 and 78 weeks. RESULTS: Intact PTH and PTH-(1-84) levels, which were 492.0 +/- 115.7 and 303.4 +/- 105.4 pg/ml, respectively, at baseline, significantly decreased at the end of the study to 268.9 +/- 121.9 (p < 0.0001) and 190.7 +/- 106.9 pg/ml (p = 0.0008), respectively. In contrast, large C-PTH fragments, which were 152.7 +/- 53.5 pg/ml at baseline, did not significantly change at 78 weeks (144.5 +/- 72.2 pg/ml, p = 0.7612). Consequently, the PTH-(1-84)/large C-PTH fragments ratio was significantly reduced from 2.25 +/- 1.31 to 1.47 +/- 0.89 (p = 0.0004). CONCLUSION: The PTH-(1-84)/large C-PTH fragments ratio reflects the change of PTH biosynthesis, processing and secretion from the parathyroid glands, and it may be a beneficial marker to evaluate the overall biological PTH action and predict bone turnover status in hemodialysis patients under intravenous vitamin D therapy. PMID- 15528945 TI - Activation of the Smad pathway in glomeruli from a spontaneously diabetic rat model, OLETF rats. AB - BACKGROUND/AIMS: Transforming growth factor-beta (TGF-beta) mediates the excess accumulation of extracellular matrix in the diabetic kidney. Smad family proteins have been identified as signal transducers for the TGF-beta superfamily. We sought to characterize the role of Smad proteins in mediating TGF-beta responses in the development of diabetic nephropathy. METHODS: We evaluated the time course of TGF-beta1 fibronectin, Smad2 and Smad3 protein expression and Smad3 activation in glomeruli from spontaneously diabetic Otsuka Long-Evans Tokushima Fatty (OLETF) rats, using immunohistochemistry and Western blot analysis. RESULTS: The glomeruli of diabetic OLETF rats showed not only accelerated activation of Smad3, but also enhanced protein expression of Smad2 and Smad3, which occurred in parallel to the increased expression of TGF-beta and fibronectin compared with glomeruli of control, Long-Evans Tokushima Otsuka (LETO) rats at 30 weeks of age. No differences were found in TGF-beta1 fibronectin, Smad2 and Smad3 protein expression and Smad3 activation in glomeruli between the two strains at 12 weeks of age when OLETF rats were not diabetic. CONCLUSIONS: The enhancement of Smad protein expression and activation may be involved in the TGF-beta signaling cascade that plays an important role in the development of diabetic nephropathy through progressive expansion of the mesangial matrix. PMID- 15528946 TI - Prospects for engineering the urinary tract. AB - Congenital abnormalities and acquired disorders can lead to organ damage or loss within the urinary tract. For reconstructive purposes, tissue engineering efforts are currently underway for virtually every type of tissue and organ within the urinary tract. Tissue engineering incorporates the fields of cell transplantation, materials science, and engineering for the purpose of creating functional replacement tissue, and requires personnel who have mastered the techniques of cell harvest, culture, expansion and transplantation, as well as polymer design. Major advances in the areas of stem cell biology, tissue engineering, and therapeutic cloning (nuclear transfer) techniques have made it possible to combine these technologies to create the comprehensive scientific field of regenerative medicine. PMID- 15528947 TI - Glomerular T cells are of restricted clonality and express multiple CDR3 motifs across different Vbeta T-cell receptor families in experimental autoimmune glomerulonephritis. AB - Experimental autoimmune glomerulonephritis (EAG) is an animal model of Goodpasture's disease which can be induced in Wistar-Kyoto (WKY) rats by a single intramuscular injection of collagenase-digested rat glomerular basement membrane (GBM) in adjuvant. This model is characterised by anti-GBM antibody production, accompanied by focal necrotising glomerulonephritis with crescent formation and glomerular infiltration by T cells and macrophages. Previous work has shown that EAG is a T-cell-dependent disease. We proposed that intraglomerular T cells might be directly involved in pathogenesis and would be oligoclonal. In this study, EAG was induced by standard methods, the kidneys perfused with saline at week 2 and week 4, and the glomeruli separated by a sieving method. Glomerular RNA was extracted and reverse transcribed. RT-PCR showed overexpression of an average of two Vbeta families in each kidney analysed. However, no predominant single Vbeta family was overexpressed in any of the experimental animals. CDR3 spectratyping of Fam-labelled PCR products showed a marked restriction involving different Vbeta families. Sequencing demonstrated multiple CDR3 motifs, each expressed in association with different Vbeta gene segments. Our results show that glomerular T cells are of restricted clonality and suggest a role for antigen-specific effector T cells in the pathogenesis of EAG. PMID- 15528948 TI - Age-dependent telomere attrition as a potential indicator of racial differences in renal growth patterns. AB - BACKGROUND: Racial differences in the predilection to salt sensitivity may arise from different renal growth patterns. To test this idea, we monitored age dependent telomere attrition rate, reflecting largely the replicative history of somatic cells, in the outer renal cortex and the inner renal medulla of African Americans and Caucasians. METHODS: Telomere length, determined by the mean length of the terminal restriction fragments (TRF), was measured in specimens from 58 African-American and 63 Caucasian males, ages 1 day to 71 years. RESULTS: In the outer renal cortex, TRF length attrition rate was significantly slower in African Americans (-0.021 +/- 0.0064 kb/year) than in Caucasians (-0.060 +/- 0.0094 kb/year) (p = 0.0007). In both ethnic groups the TRF length attrition rate was slower in the inner medulla than in the outer renal cortex, but without significant racial differences. CONCLUSIONS: The proximal tubule is the most abundant nephron structure in the outer renal cortex. Less proliferative growth of proximal tubular cells in kidneys from African Americans may be one factor explaining the slower age-dependent telomere attrition rate in the outer renal cortex of African Americans than in Caucasians. PMID- 15528949 TI - Interaction between the polyol pathway and non-enzymatic glycation on mesangial cell gene expression. AB - BACKGROUND/AIMS: Both activation of the polyol pathway and enhanced non-enzymatic glycation have been implicated in the pathogenesis of diabetic glomerulopathy. We investigated the interaction between these two pathways using normal mesangial cells (MCs) and transgenic (TG) MCs with elevated aldose reductase (AR) activity. METHODS: TG mice with expression of the human AR (hAR) gene in kidney MCs were established. Mouse glomeruli and primary cultures of MCs from hAR TG and wild type (WT) mice were studied regarding the changes in AR activity, transforming growth factor-beta1 (TGF-beta1) and type IV collagen mRNA and protein levels, in response to BSA modified by advanced glycation end-products (AGE-BSA). RESULTS: Ex vivo addition of AGE-BSA increased AR activity, TGF-beta1 and type IV collagen mRNA levels in both WT and TG glomeruli, with greater rise in TG glomeruli. These increments were attenuated by zopolrestat, an AR inhibitor. In cultured MCs, AGE BSA enhanced AR activity, TGF-beta(1) and type IV collagen mRNA and protein levels both in WT and TG MCs, again with greater increases in TG MCs. The AGE induced enhancement in TGF-beta1 and type IV collagen expression were suppressed by either zopolrestat or transfection with an AR antisense oligonucleotide. CONCLUSION: These data suggest that the activation of the polyol pathway by AGEs, more marked in genetic conditions with increased AR activity, may contribute to the pathogenesis of diabetic glomerulopathy, through enhancing mesangial cell expression of TGF-beta1 and type IV collagen. PMID- 15528950 TI - Effects of leptin on intracellular calcium concentrations in isolated porcine somatotropes. AB - Leptin, the product of the obese gene, is a protein that is secreted primarily from adipocytes. Leptin can influence the function of the pituitary gland through its action on the hypothalamus, but it can also directly act at the level of the pituitary gland. The ability of leptin to induce an increase in intracellular Ca2+ concentration ([Ca2+]i) in somatotropes was examined in dispersed porcine pituitary cells using a calcium imaging system. Somatotropes were functionally identified by the application of human growth hormone releasing hormone. Leptin increased [Ca2+]i in porcine somatotropes in a dose-dependent manner. The application of 100 nM leptin for 3 min did not have a significant effect on [Ca2+]i, while a 3-min application of 1 microM leptin increased [Ca2+]i in about 50% of the somatotropes (p < 0.01). The application of a second leptin challenge (1 microM) evoked a response in only 18% of the observed somatotropes. The stimulatory effect of leptin was abolished in low calcium saline and blocked by nifedipine, an L-calcium channel blocker, suggesting an involvement of calcium channels. Pretreatment of the cultures with AG 490, a specific Janus kinase inhibitor, and with SB 203580, a mitogen-activated protein kinase (MAP kinase) inhibitor, abolished the increase in [Ca2+]i evoked by leptin. In the presence of N(omega)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, the magnitude of the increase in [Ca2+]i evoked by 1 microM leptin was not significantly changed. However, in the presence of L-NAME only 24% of the somatotropes responded to leptin, while in parallel control cultures 70% of the somatotropes responded to leptin. These results imply an involvement of Janus kinase/signal transducer and activator or transcription, MAP kinase and NOS signaling pathways in the stimulatory effect of leptin on porcine somatotropes. PMID- 15528951 TI - Vasopressin stimulates ventromedial hypothalamic neurons via oxytocin receptors in oxytocin gene knockout male and female mice. AB - A wealth of neuropharmacological data demonstrates that oxytocin (OT) actions in the mammalian forebrain support a wide variety of affiliative behaviors and repress aggressive behaviors. Based on that literature, it was expected that reproductive and affiliative behaviors would be vastly decreased and aggression markedly increased in OT gene knockout (OTKO) mice. The initial publications reporting the behaviors of these mice did not include such phenotypes. Here, we compared single-unit activities recorded from the ventromedial hypothalamus in tissue slices of male and female OTKO mice and their wild-type littermate to test two hypotheses about OT functional genomics. First, we proposed that in OTKO mice, a very similar 9-amino-acid neuropeptide, arginine vasopressin (a likely gene duplication product), can 'cross over' and compensate for the lack of OT. This hypothesis was confirmed in both males and females. Further, we proposed that because of the lifelong absence of OT in OTKO, OT receptors would be more sensitive to OT in the knockout animals. We tested this idea in males and found that it was correct. Thus, an answer to the 'OTKO paradox' is put forth, with implications for OT-sensitive behaviors in a variety of species. PMID- 15528952 TI - The role of vascular endothelial growth factor in the kidney in health and disease. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen, angiogenic factor and enhancer of vascular permeability. Expressed in the epithelial cells of the developing glomerulus and tubular epithelium, VEGF plays an important role in the development and maintenance of the early vasculature of the kidney. Here, we review the available literature regarding the expression and function of VEGF both in the developing and healthy adult kidney. Furthermore, we highlight how VEGF expression is altered in the diseased kidney and how this modulated expression may impact on and reflect underlying functional changes occurring during the disease process. As discussed, many controversial issues remain, particularly concerning the role of VEGF in the diseased kidney. That VEGF has been proposed as a potential future therapeutic target for the management of some renal diseases requires first that the precise role of VEGF in the normal kidney and various renal pathologies be further and more clearly defined. PMID- 15528953 TI - Inhibitory effects of ligustrazine, a modulator of thromboxane-prostacycline nitric oxide balance, on renal injury in rats with passive Heyman nephritis. AB - AIMS: To explore the effects of ligustrazine on proteinuria, urinary TxB2 (metabolism of thromboxane A2, TxA2) and 6-keto-PGF1alpha (metabolism of prostacyclines I2, PG I2), glomerular inducible nitric oxide(NO) synthase (iNOS) mRNA, urinary NO3-/NO2- (decomposing products of NO) and pathological changes in rats with passive Heymann nephritis (PHN). METHODS: A rat PHN model was induced by intravenous injection of rabbit anti-rat renal tubular antigen (Tub-Ag) antiserum, and ligustrazine was given intraperitoneally into PHN rats every 2 days for 1-5 weeks. Then, proteinuria, urinary TxB2 and 6-keto-PGF1alpha, glomerular iNOS mRNA, and urinary NO3-/NO2- were measured by sulfosalicylic acid, radioimmunoassay (RIA), Northern blot and nitric acid reductase methods, respectively. Moreover, the damage to the renal tissue of the rats was observed under light and electron microscopy and immunofluorescence (IF). RESULTS: The urinary TxB2 in PHN rats was significantly higher than that in control rats, but the PHN rats treated with ligustrazine had significantly less proteinuria, urinary TxB2 and tissue lesions, and more urinary 6-keto-PGF(1alpha), glomerular iNOS mRNA and urinary NO2-/NO3- than the PHN rats without the administration of ligustrazine. CONCLUSION: These data indicate that ligustrazine has inhibitory roles on the glomerular injury of PHN rats, which may associate with modulating the balance of TxA2-PGI2 and elevating synthesis of NO to a certain extent. PMID- 15528954 TI - Inducible nitric oxide synthase activity and expression in liver and hepatocytes of diabetic rats. AB - Inducible nitric oxide synthase (iNOS) is expressed by the liver in a number of physiological and pathophysiological conditions. The aim of this study was to investigate the relationship between the diabetic state, iNOS and oxidative stress in the rat liver and isolated hepatocytes. Hepatic iNOS expression and activity was measured in both healthy and streptozotocin-induced diabetic rats and determined in hepatocytes in the presence and absence of insulin. Cu/Zn superoxide dismutase (SOD) and phosphatidylinositol-3-kinase (PI3K) were also measured. In a separate experiment lasting 3 weeks, diabetic rats received either no treatment, two daily injections of insulin or aminoguanidine in the drinking water. Diabetes led to increased activity (45%) and expression (70%) of liver iNOS, an effect that was attenuated by insulin treatment both in vitro and in whole animals. Hepatocyte iNOS expression increased by 56%. Hepatic SOD expression was elevated in the diabetic state, but activity levels were similar to healthy controls. Insulin treatment in vivo led to increased enzyme activity but expression was not modified. Levels of PI3K protein were significantly lower in diabetic rats while insulin treatment markedly increased expression. Aminoguanidine did not inhibit hepatic iNOS in this study. Glycemic control via insulin administration was able to downregulate enhanced hepatic iNOS activity and expression in the liver observed in the diabetic state and improve SOD activity, responses that can potentially reduce the free radical damage associated with diabetes. PMID- 15528955 TI - Nonhabitual seizures in patients with implanted subdural electrodes. AB - The implantation of subdural electrodes has been widely employed in the invasive monitoring of patients with medically refractory epilepsy. The use of subdural electrodes, though, has been associated with rare but occasionally troublesome complications. We report the occurrence of nonhabitual seizures after implanting subdural grid electrodes. Among 57 patients diagnosed with medically refractory epilepsy who were evaluated in our department over a 12-month period, 21 patients underwent craniotomy for subdural grid/strip electrode implantation. Subdural grids and strips (AdTech, Racine, Wisc., USA) were used for continuous video EEG monitoring. In 3 patients, during subdural monitoring, consistent nonhabitual seizure activity was recorded. This was both clinically and electrographically different than the patients' habitual seizures. The patients' nonhabitual seizures disappeared postoperatively after removing the implanted electrodes. The occurrence of nonhabitual seizures, though quite rare, could lead to mislocalization of an epileptogenic focus. This complication might be the result of direct mechanical cortical irritation or chemical irritation caused by blood breakdown products. The occurrence of nonhabitual seizures comes to add itself to the existing list of complications associated with employment of subdural electrodes for invasive monitoring. PMID- 15528956 TI - How do dermatological vehicles influence the horny layer? AB - Emulsifying agents in aqueous solution lead to dehydration of the horny layer of the epidermis and damage to the barrier. This is only partly true if emulsifying agents are constituents of emulsions. Water-in-oil (W/O) emulsions cause an improvement in the hydration of the horny layer and barrier function. In the case of an additional effect of wash solutions, a partial or complete abolition of the drying out and barrier-damaging effect of the wash solution results. Oil-in-water (O/W) emulsions without glycerol have no effect on the moisture of the horny layer or indeed increase it, but they lead to considerable damage to the barrier and irritation. In wash tests, they do not induce any protective effect. With micro-emulsions this is still more the case and, moreover, they can also lead to exsiccation. As a result of the addition of glycerol, the barrier-damaging effect can be abolished. Also in stress tests with wash solutions, the damage to the horny layer is reduced by glycerol-containing O/W emulsions. Whereas the penetration-promoting effect of O/W emulsions without glycerol is best, only W/O emulsions or glycerol-containing O/W emulsions are suitable for atopic dermatitis. A hydrating effect on the stratum corneum was also found in a propylene glycol ointment. PMID- 15528957 TI - Topical delivery of retinyl ascorbate co-drug. 2. Comparative skin tissue and keratin binding studies. AB - Retinyl ascorbate (RA-AsA), an ester co-drug of vitamins A (RA) and C (AsA), is proposed as a topical antioxidant/cell division regulator for reducing UV-induced generation of free radicals and disrupted dermal cell growth. The efficacy of dermatological agents is influenced by their retention within the skin, which is increased by the interaction with skin components. Keratin is the major protein (approximately 95%) in the skin, and this paper reports the binding of RA-AsA, RA, AsA, retinol, ascorbic acid palmitate and retinol palmitate to three tissues human callus, pig ear skin and bovine horn keratin. Tissue samples were incubated with solutions of compounds and the uptake measured as the ratio of bound/free compound at equilibrium. Binding to keratin was assessed using delipidised tissue, and was much higher for the polar compounds, suggesting dipolar/H-bonding interaction. Binding strength was ranked as human > porcine > bovine, but there was no distinction for highly lipophilic compounds. The binding characteristic of native tissues was complicated by lipid content of the tissues. There seemed to be a dual effect. The binding of very lipophilic materials increased with lipid content, implying that a substantial amount is dissolved in the lipid matrix. For highly polar AsA, lipid content decreased the binding, suggesting that the lipid reduced the strong polar interactions with skin protein/keratin. PMID- 15528958 TI - Effects of L-Ornithine on metabolic processes of the urea cycle in human keratinocytes. AB - The complex metabolic function of L-ornithine has led to a great interest in studying its pharmacotherapeutic potential. L-Ornithine is known to be crucial for the metabolism of keratinocytes, especially in the synthesis of urea, polyamines and precursors of collagen synthesis. In this study, we investigated in vitro the cytotoxicity of L-ornithine, and its influence on urea synthesis and arginase expression in primary human keratinocytes. L-Ornithine (> or =1 mM) induced a decrease in the de novo urea synthesis of keratinocytes and an increase (> or =10 mM) in the expression of the urea-generating enzyme arginase. Up to 20 mM, L-ornithine showed no cytotoxic potential, whereas higher concentrations induced apoptosis in keratinocytes in a concentration- and time-dependent manner. PMID- 15528959 TI - Transdermal iontophoresis of insulin. VI. Influence of pretreatment with fatty acids on permeation across rat skin. AB - The delivery of large peptides through the skin poses a significant challenge, and various strategies are under active investigation for enhancing the transdermal permeation. For large peptides, it is difficult to achieve significant permeation using iontophoresis alone. Hence a combination of fatty acids with iontophoresis was hypothesized to result in higher enhancement than achieved with either of them alone. Saturated fatty acids and cis unsaturated fatty acids were studied in combination with iontophoresis using excised rat skin. The skin was pretreated for 2 h with an ethanolic (EtOH) solution of 5% w/v or v/v fatty acids, namely lauric acid (LA), oleic acid (OA), linoleic acid (LOA) and linolenic acid (LLA), followed by either passive or iontophoretic permeation (0.5 mA/cm2 for 6 h). Fourier transform infrared spectroscopy (FT-IR) was used to investigate the biophysical changes on treatment with fatty acid/EtOH or neat fatty acid, mainly focusing on the infrared region at 2,920, 1,710 and 1,720 cm( 1). Unsaturated fatty acids showed higher enhancement than LA, and the enhancement increased with the number of double bonds. On the other hand, in the presence of iontophoresis, LA/EtOH showed the highest enhancement. Neat LOA did not show any significant difference (p > 0.05) compared to the LOA/EtOH combination. FT-IR studies revealed that fatty acids act by interacting with the skin lipids. All the fatty acids showed synergistic enhancement when combined with iontophoresis. The flux enhancement was highest with LA, which in the presence of iontophoresis showed 20 times enhancement of insulin flux in comparison to passive flux and 9 times enhancement as compared to iontophoresis alone. Flux enhancement of unsaturated fatty acids was in the following decreasing order LOA > OA > LLA. PMID- 15528960 TI - Ex vivo cutaneous absorption assessmentof a stabilized ascorbic acid formulation using a microdialysis system. AB - BACKGROUND: Reactive oxygen species generated by ultraviolet light result in photocarcinogenic and photoaging changes in the skin. Antioxidants protect the skin from these insults. OBJECTIVE: The aim of this study was to determine the ex vivo ascorbic acid penetration and its degradation in the skin after its topical application from an 8% new formulation. METHOD: Ascorbic acid was applied to human skin fragments. Ascorbic acid and its metabolites were collected by microdialysis and assessed by gas chromatography mass spectrometry. RESULTS: After topical application of the new formulation, the ascorbic acid level achieved was 8.5% higher than [corrected] times the normal tissue value. This high ascorbic acid dermal concentration remained constant if a topical application was made every 8 h. No degradation of ascorbic acid was detected. CONCLUSION: Ascorbic acid penetrates rapidly after its topical application. The persistent reservoir of ascorbic acid provides an important and attractive photoprotection strategy. PMID- 15528961 TI - Basal cell adhesion molecule is inversely associated with apoptosis, but plays a limited role for protection against apoptotic stimuli. AB - Basal cell adhesion molecule (B-CAM) is strongly upregulated in epithelial skin cancer cells in vivo and in vitro. We have tested here whether B-CAM is (1) inversely associated with or (2) functionally involved in apoptosis. Towards this end, B-CAM expression was assessed in HaCaT transfectants overexpressing murine Bcl-2 and untransfected HaCaT cells exposed to various proapoptotic stimuli. In another series of experiments, we overexpressed B-CAM in HaCaT cells and different fibroblast lines, and stimulated various apoptotic pathways in the transfectants and control cells. In addition, apoptosis was assessed after an antibody-mediated B-CAM blockade. We could demonstrate that expression of B-CAM is inversely associated with the susceptibility of cells to apoptosis. However, overexpression or antibody- mediated inhibition of B-CAM had only limited functional effects on cellular apoptosis. PMID- 15528962 TI - Formulation of an HPMC gel drug reservoir system with ethanol-water as a solvent system and limonene as a penetration enhancer for enhancing in vitro transdermal delivery of nicorandil. AB - The objective of the present study was to formulate a hydroxypropyl methylcellulose (HPMC) gel drug reservoir system with ethanol-water as a solvent system and limonene as a penetration enhancer for enhancing the transdermal delivery of nicorandil so as to develop and fabricate a membrane-moderated transdermal therapeutic system (TTS). The in vitro permeation of nicorandil was determined across rat abdominal skin from a solvent system consisting of ethanol or various proportions of ethanol and water. The ethanol-water (70:30 v/v) solvent system that provided an optimal transdermal permeation was used in formulating an HPMC gel drug reservoir system with selected concentrations (0% w/w, 4% w/w, 6% w/w, 8% w/w or 10% w/w) of limonene as a penetration enhancer for further enhancement of transdermal permeation of nicorandil. The amount of nicorandil permeated in 24 h was found increased with an increase in the concentration of limonene in the drug reservoir system up to a concentration of 6% w/w, but beyond this concentration there was no further increase in the amount of drug permeated. The flux of nicorandil was 370.9 +/- 4.2 microg/cm2 x h from the drug reservoir system with 6% w/w of limonene, which is about 2.6 times the required flux to be obtained across rat abdominal skin for producing the desired plasma concentration for the predetermined period in humans. The results of a Fourier Transform Infrared study indicated that limonene enhanced the percutaneous permeation of nicorandil by partially extracting the stratum corneum lipids. It is concluded that the HPMC gel drug reservoir system prepared with a 70:30 v/v ethanol-water solvent system containing 6% w/w of limonene is useful in designing and fabricating a membrane-moderated TTS of nicorandil. PMID- 15528963 TI - Expression of sigma 1 receptor in human breast cancer. AB - The sigma 1 receptor (S1R) represents a unique drug-binding site that is distinct from any other receptors. We examined S1R expression in human breast cancer and assessed the activity of S1R ligands in breast cancer cell lines. One-hundred nine breast specimens from normal breast, benign breast disease and cancer were examined with immunohistochemistry or RT-PCR and six different cell lines were also evaluated. S1R mRNA overexpression was detected in 64% of breast cancers compared to normal breast tissue. Immunohistochemistry showed positive epithelial cell staining in 60% of invasive and 41% of in situ cancers, 75% of ductal hyperplasia and in 33% of normal breast. The pattern of expression was more diffuse in invasive breast carcinoma compared to other conditions (p = 0.02). S1R expression was neither a prognostic nor a predictive factor for efficacy of adjuvant chemotherapy but the study only included 58 cancer patients and therefore the statistical power is limited. MDA-MB-361, MDA-MB-435, BT20 and MCF7 cells all expressed S1R protein by Western blot. The non-specific S1R ligands haloperidol, reduced haloperidol and progesterone produced a dose-dependent inhibition of the growth at high (>10 microM) concentrations. Reduced haloperidol also showed additive cytotoxic effects when combined with doxorubicin, vinorelbine , paclitaxel and docetaxel in vitro. The S1R-specific ligand, SKF 10047 demonstrated the least growth inhibitory activity and showed no interaction with chemotherapy. These results demonstrate that some normal and most neoplastic breast epithelial cells and cell lines commonly express S1R. High concentrations of haloperidol inhibit the growth of these cells and potentiate the effect of chemotherapy in vitro. PMID- 15528964 TI - Immunocytochemical detection of epithelial cells in the bone marrow of primary breast cancer patients: a meta-analysis. AB - PURPOSE: The immunocytochemical detection of epithelial cells in the bone marrow (BM) of breast cancer (BC) patients has been shown to have prognostic importance in several studies. We conducted a systematic review of the literature and a meta analysis to assess the specificity, rate of positivity and correlation with known prognostic variables as well as with disease free (DFS) and overall survival (OS). DESIGN: We performed a systematic review from the published literature. We included studies with at least 20 previously untreated BC patients as well as non BC controls that evaluated the presence of epithelial cells in BM using immunocytochemistry (IC). RESULTS: We identified 14 eligible studies with 3253 BC patients and 532 controls. Our results showed that IC has an overall rate of positivity of 31% (95% CI: 30-33%), and specificity of 96% (compared to normal patients) (95% CI: 0.94-0.98). We observed significant direct correlations between IC BM positivity and the primary tumor's presence with expression of estrogen receptors, larger size and higher histologic grade as well as with the presence of more than 3 positive axillary lymph nodes. At 5 years of follow up IC positivity correlated directly with a lower DFS (relative risk=1.60; 95% CI: 1.39 1.83) and OS (relative risk=1.73; 95% CI: 1.29-2.31). The prognostic impact of BM positivity seemed to decrease with time. CONCLUSIONS: We conclude that the finding of epithelial cells in BM of BC patients correlates with several known prognostic factors and has adverse impact in the DFS and OS of these patients that seems, however, to decrease with time. PMID- 15528965 TI - The potential role of breast conservation surgery and adjuvant breast radiation for adenoid cystic carcinoma of the breast. AB - PURPOSE: Adenoid cystic carcinoma (ACC) of the breast is a rare breast cancer variant and optimal management is unclear. A review of this unusual tumour was performed at our Institution, to assess the role of breast conservation in the management of this disease. METHODS AND MATERIALS: A review of all cases of ACC of breast (1960-2000) treated at Princess Margaret Hospital (PMH) was undertaken. Information was collected on age at diagnosis, presenting features, tumour size and treatment modalities. Treatment outcomes were evaluated. RESULTS: Eighteen female and one male patient were identified. Median age at diagnosis was 58 years (range 35-76 years). Four patients had lymph-node positive disease at presentation; the single male patient presented with metastatic disease. Surgery was either a lumpectomy (10 cases) or a simple, radical or modified radical mastectomy (9 patients). Nine of 19 patients received adjuvant radiotherapy (RT). The median follow-up time was 14 years; the recurrence rate at 10 years was 31% (95% CI 7-54%) with a range in time of recurrence from 2.3 to 11.9 years. Seven recurrences were identified (4 local, 1 regional, 2 metastatic). Two of these patients developed metastatic spread and died. Six of the 19 cases went on to develop second malignancies of whom four died. Among the 18 female patients, the 10-year overall (OS), cause-specific (CSS), and relapse free survival (RFS) rates were 75, 100, and 46% respectively. CONCLUSIONS: ACC of the breast has a relatively prolonged natural history, and responds well to conservative management at presentation, with good outcome, even following local recurrence. PMID- 15528966 TI - Quality of life in randomized trials of cytotoxic or hormonal treatment of advanced breast cancer. Is there added value? AB - BACKGROUND: Since most advanced cancers are still incurable, oncologic clinical research pays considerable attention to palliation, increasingly valuing subjective measures of outcome such as quality of life (QoL). We reviewed randomised clinical trials (RCT) of cytotoxic or hormonal treatments in advanced breast cancer (ABC), published before December 2003, to evaluate the methodological quality of QoL assessment and assess its added value (over classical clinical endpoints (CCE), i.e. survival, response, time to progression, toxicity) in the choice of the best treatment option. METHODS: RCTs were classified according to treatment characteristics and the CCEs. A descriptive analysis was based on the methodological aspects of QoL assessment and the clinical value of QoL findings was judged by counting the frequency of reporting in the study abstracts and the assessment of QoL combined with CCEs. RESULTS: We retrieved 33 eligible RCTs (10,791 patients); only 20 reported the number of patients considered in QoL principal analysis and only 69% of randomized patients were included in such analyses. A total of 17 different QoL questionnaires were used, 11 only once. QoL assessment lasted from less than 12 weeks to progression, and timing of questionnaires from 2 to 12 weeks. Compliance rates were 85.7% for baseline forms and 67% for overall assessment, but this information was available for only 18 and 20 trials, respectively. Wide variability emerged in analysis strategies and statistical approaches. QoL findings were reported in 12 study abstracts (37% of patients). Eight studies reported a significant difference in QoL scores but since QoL data often failed to parallel the clinical findings (e.g. better QoL scores were reported in two of 17 trials with better CCEs and in six of 20 with significant differences in toxicity profiles), the QoL added value was difficult to ascertain and, on the whole, only moderate. CONCLUSION: In ABC trials, QoL assessment added relatively little value to CCEs in helping select the best treatment option, apparently largely because of sub-optimal methodological standards. PMID- 15528967 TI - Liposomal 4-hydroxy-tamoxifen: effect on cellular uptake and resulting cytotoxicity in drug resistant breast cancer cells in vitro. AB - The study was performed to investigate the potency of liposomal formulations to reduce or to overcome tamoxifen resistance. Therefore, estrogen receptor (ER) positive MCF-7 and ER deficient NCI/ADR cells with different sensitivity to tamoxifen were used. Instead of the parent compound, 4-hydroxytamoxifen (HT) was used as drug, because this metabolite is the most cytotoxic derivative in vitro. Liposomes further contained the membrane-active alkylphospholipid octadecyl-(1,1 dimethyl-piperidino-4-yl)-phosphate (OPP). Cellular uptake of HT during 3-24 h was determined by high performance thin layer chromatography technique (HPTLC). Free HT was taken up time independently by the two cell lines at 1.5-2.1 g HT/g cellular phosphate. Liposomal HT was taken up at a significantly higher degree than free HT in both cell lines, but the uptake was delayed in the sensitive MCF 7 cell line with the highest concentration detected after 24 h (3.5 g/g). Oppositely, the highest amount in the 'resistant' line (2.3 g/g) was already measured after 3 h in NCI/ADR cells. It successively decreased with incubation time. The faster uptake of liposomal HT by the NCI/ADR cells correlated with a stronger and earlier destruction of resistant NCI/ADR cells whereas the sensitive MCF-7 cells were mainly inhibited in their proliferation. Cytolytic effects were observed in both cell lines after extended incubation periods. The combination of HT with an alkylphospholipid further enhanced the cytotoxicity of the formulation. The IC50 in the NCI/ADR cells could be significantly reduced by liposomes combining both drugs to 15.1 microM compared with the IC50 of the free drugs (HT:28.9 microM; OPP: 36.8 microM). It is assumed that the enhanced and accelerated uptake of liposomal HT in the cell line with relative drug resistance can increase the intracellular bioavailability of HT. The results of this study demonstrated that liposomes with encapsulated antiestrogen have a superior cytotoxic effect in resistant breast cancer cells. That coincided with the enhanced therapeutic effect of these vesicles observed in vivo. PMID- 15528968 TI - Induction of apoptosis by AN-152, a cytotoxic analog of luteinizing hormone releasing hormone (LHRH), in LHRH-R positive human breast cancer cells is independent of multidrug resistance-1 (MDR-1) system. AB - OBJECTIVE: More than 50% of human breast cancers express receptors for luteinizing hormone-releasing hormone (LHRH-R). These receptors can be used for targeted chemotherapy with agents like AN-152, in which doxorubicin is linked to analog [D-Lys6]LHRH. We compared the effects of AN-152 and doxorubicin in human breast cancer cells. METHODS: MCF-7, T47D, HCC-70 and ZR-75-1 cells were analysed for expression of LHRH-R using RT-PCR, immunostaining and flow cytometry. Apoptosis and expression of MDR-1 gene product Pgp were measured by flow cytometry. Cleavage of doxorubicin from AN-152 by serum carboxylesterase (CE) was inhibited by DFP. RESULTS: In MCF-7, T47D and HCC-70 cells we found cell surface expression of LHRH-R. In ZR-75-1 cells only sparse surface expression was found. In HCC-70 cells, induction of apoptosis by AN-152 was stronger than by doxorubicin in medium containing fetal calf serum (FCS). Pretreatment with DFP increased AN-152-induced apoptosis in LHRH-R positive MCF-7 and HCC-70 cells and reduced apoptosis in ZR-75-1 cells. In serum-free medium apoptosis induced by AN 152 was stronger than that induced by doxorubicin in HCC-70, T47D and MCF-7 cells, but weaker in ZR-75-1 cells. In medium containing FCS, both AN-152 and doxorubicin induced surface expression of MDR-1 gene product Pgp, but the effect of AN-152 was weaker. Pgp-expression induced by AN-152 was inhibited by pretreatment with DFP. AN-152 did not induce Pgp-expression in serum-free medium. CONCLUSION: The cytotoxic LHRH analog AN-152 induces apoptosis independent of MDR 1 in LHRH-R positive breast cancer cells. The efficacy and/or specificity of AN 152 is improved by suppression or absence of CE activity. PMID- 15528969 TI - Is expert breast pathology assessment necessary for the management of ductal carcinoma in situ ? AB - BACKGROUND: Current guidelines include a recommendation that a pathologist with expertise in breast disease review all ductal carcinoma in situ (DCIS) specimens due to the presence of significant variability in pathologic reporting of DCIS. The objective of this study was to evaluate the completeness and accuracy of pathologic reporting of DCIS over the past decade and to determine the current impact of expert breast pathology assessment on the management of DCIS. METHODS: All patients with a diagnosis of DCIS referred to a single regional cancer centre between 1982 and 2000 have been reviewed. Inter-observer variability between initial and secondary reports has been evaluated using kappa statistics. For each case, the Van Nuys Prognostic Index (VNPI) using pathologic data obtained from the initial and reviewed pathology reports were compared. The impact of expert breast pathology on risk assessment and treatment was determined. RESULTS: 481 individuals with DCIS were referred and pathology review was performed on 350 patients (73%). Inter-observer agreement was high for the main pathologic features of DCIS. From 1996 to 2000, secondary pathology assessments lead to a change in the assessment of local recurrence risk in 100 cases (29%) and contributed to a change in treatment recommendation in 93 (43%) cases. CONCLUSION: Expert breast pathology assessments continue to be necessary in the management of DCIS. PMID- 15528970 TI - Loss of heterozygosity at BRCA2 in a ductal carcinoma in situ and three invasive breast carcinomas in a family with a germline BRCA2 mutation. AB - We have examined a family with a germline BRCA2 mutation in three cases of invasive breast cancer and one case of ductal carcinoma in situ (DCIS). Loss of heterozygosity (LOH) has been demonstrated at the BRCA2 locus in all cases. This result may suggest that the germline mutation in BRCA2 is the initiating step of DCIS and support the theory that DCIS is a precursor of invasive breast carcinoma in hereditary breast cancer. PMID- 15528971 TI - Mammary tumorigenesis in growth hormone deficient spontaneous dwarf rats; effects of hormonal treatments. AB - This study was carried out to investigate mammary tumorigenesis in growth hormone (GH) deficient spontaneous dwarf rats (SDR). At 50-60 days of age, the rats were divided into five groups. Group 1 received bovine (b) GH (prolonged release formulation) administered at a dose of 40-50 mg/kg body wt. in 50 microl weekly injections; group 2 received recombinant human insulin-like growth factor-I (IGF I) at a dose of 1 mg/kg body wt./day administered via osmotic pumps; animals in group 3 were fitted with subcutaneous silastic capsule containing 30 microg 17 beta-estradiol (E2) plus 30 mg progesterone (P4), replaced every 2 months; group 4 received both bGH and E2 plus P4 treatments at the same doses as above, and control animals (group 5) received sham treatments (vegetable oil injection, silastic capsules containing cellulose). After 1 week of treatment, all animals were injected intraperitoneally with the carcinogen N-methyl-N-nitrosourea (MNU) at a dose of 50 mg/kg body wt. Other groups of animals, receiving identical hormonal treatment to those exposed to MNU, were treated for 10 days only and then sacrificed for assessment of circulating concentrations of hormones and mammary gland characteristics at the time of carcinogen exposure. The hormonal treatments of the animals exposed to the MNU were continued for an additional 20 weeks and mammary tumor development monitored by weekly palpation and tumors collected as necessary. The rats were weighed weekly. At the end of the treatment period, all animals were sacrificed and remaining tumors were collected. Rats in all groups continued to gain weight throughout the experimental period, but the largest weight gain was see in animals receiving GH either alone or with E2 and P4. Animals treated with IGF-I also gained weight compared to controls, but this weight gain was less than that seen in GH-treated rats. GH treatment alone increased mammary tumor incidence from 4.8% in controls to 100%. Average tumor load and latency in the GH-treated rats were 7.0 +/- 0.8 tumors/tumor-bearing rat (mean +/- SEM) and 57.3 +/- 2.7 days (mean +/- SEM), respectively. As in intact Sprague-Dawley rats, approximately 90% of the tumors that developed in the GH treated rats were ovarian dependent for growth. IGF-I treatment also increased mammary tumor development to 62.5%. Average tumor load and latency in the IGF-I treated rats were 1.6 +/- 0.4 tumors/tumor-bearing rat (mean +/- SEM) and 96.2 +/ 14.5 days (mean +/- SEM), respectively. However E2 + P4 treatments did not significantly alter tumorigenesis and, surprisingly, simultaneous treatment with E2 + P4 and GH obliterated the GH-stimulated increase in tumor development. Prolactin (PRL) did not appear to influence mammary tumorigenesis in the SDRs, as untreated SDRs had significantly elevated serum concentration of PRL as compared with normal Sprague-Dawley (SD) rats, whereas GH-treated SDRs had PRL levels similar to that of normal SD rats. No obvious structural characteristics were associated with high or low susceptibility to mammary tumorigenesis, as assessed by mammary gland whole mounts from the different animal groups sacrificed at the time of carcinogen administration. Enhanced expression of the extracellular signal-regulated kinase 1/2 (ERK1/2), and activation (phosphorylation) of ERK1/2 were associated with an increase in mammary tumorigenesis. Similarly, the expression of the estrogen receptor-alpha (ER alpha) was significantly elevated in animal groups with the highest susceptibility to tumorigenesis, whereas the levels of cyclin D1 expression were not related to mammary tumorigenesis. PMID- 15528972 TI - Breast cancer aggressiveness and racial disparity. PMID- 15528975 TI - Investigation of HMN-176 anticancer activity in human tumor specimens in vitro and the effects of HMN-176 on differential gene expression. AB - HMN-176, (E)-4-(2-[2-(N-[4-methoxybenzene-sulfonyl]amino)phenyl]ethenyl) pyridine 1-oxide, is a stilbene derivative which inhibits mitosis without significant effect on tubulin polymerization and displays potent cytotoxicity against a variety of human tumor cell lines. The present study evaluated the activity profile of the antineoplastic agent HMN-176 in an ex-vivo soft agar cloning assay (human tumor colony-forming assay) in a panel of 132 human tumor specimens under 14-day continuous exposure at 0.1, 1.0, and 10.0 microg/ml. Thirty percent of specimens in the different treatment groups (39/132 in 0.1 and 1.0 test groups; 40/132 in 10.0 test group) were assessable, falling within the negative and positive control parameters. At these dose levels, responses were observed in 32% (11/34), 62% (21/34), and 71% (25/35) of assessable specimens, respectively. HMN 176 demonstrated activity towards 75% of the breast cancer specimens (6/8) treated with 1.0 microg/ml, 67% of non small-cell lung (4/6) and 57% of ovarian (4/7) cancer specimens treated with 10 microg/ml. Low levels of cross-resistance to cisplatin, cyclophosphamide, 5-fluorouracil, and etoposide were also observed. There was a positive relationship between HMN-176 concentration and effect, demonstrating greatest overall activity at 10.0 microg/ml. Evaluation of differential gene expression in drug-sensitive (A2780) and drug-resistant (A2780cp) ovarian carcinoma cell lines exposed to 0.1 microg/ml HMN-176 up to 48 h using cDNA microarrays with 1,154 known human genes revealed significant drug effects on tumor associated genes, including upregulation of tissue inhibitor matrix metalloproteinases gene (TIMP) in both cell lines, suggesting that HMN-176 could potentially overcome tumor drug resistance. In conclusion, in vitro responses demonstrate efficacy at pharmacologically relevant concentrations, which suggests that HMN-176 deserves further evaluation in clinical trials. PMID- 15528977 TI - Inhibition of lymphocyte activation and function by the prenylation inhibitor L 778,123. AB - Prenylated Ras GTPases transduce signals from the T cell receptor, CD28 costimulatory receptor and IL-2 receptor. Since signals from these receptors mediate T cell activation, proliferation and survival, we hypothesized that the prenylation inhibitor L-778,123 would impart immunomodulation. The effect of L 778,123 on T cell activation (CD71 or CD25 surface expression) was determined by flow cytometry. Peripheral blood mononuclear cell (PBMC) proliferation in the presence of L-778,123 and/or cyclosporine (CsA) was determined by [3H]thymidine incorporation. The ability of L-778,123 to inhibit IL-2 receptor signaling was investigated by measuring IL-2 induced proliferation in CTLL-2 cells and IL-2 prevention of apoptosis in activated human PBMC. L-778,123 inhibited lectin induced expression of CD71 and CD25 with IC50's of 6.48 +/- 1.31 microM and 84.1 +/- 50.0 microM, respectively. PBMC proliferation was inhibited by L-778,123 with an IC50 of 0.92 +/- 0.23 microM, and addition of CsA did not increase the potency. L-778,123 did not inhibit IL-2 and IFN-gamma production by T cells. L 778,123 abrogated IL-2 induced proliferation of CTLL-2 cells with an IC50 of 0.81 +/- 0.44 microM. However, L-778,123 minimally reversed the prosurvival effect of IL-2 in activated lymphocytes. IL-2 ligand and receptor production during T cell activation are relatively unaffected by L-778,123. However, the activation and proliferative effects of IL-2 on T cells are potently blocked by L-778,123. These results reveal a selective blockade of the IL-2 cytokine axis distal to the IL-2 receptor by the L-778,123 and warrant evaluation of prenylation inhibitors in treating transplant rejection and autoimmune diseases. PMID- 15528976 TI - Possible synergistic prostate cancer suppression by anatomically discrete pomegranate fractions. AB - We investigated whether dissimilar biochemical fractions originating in anatomically discrete sections of the pomegranate (Punica granatum) fruit might act synergistically against proliferation, metastatic potential, and phosholipase A2 (PLA2) expression of human prostate cancer cells in vitro . Proliferation of DU 145 human prostate cancer cells was measured following treatment with a range of therapeutically active doses of fermented pomegranate juice polyphenols (W) and sub-therapeutic doses of either pomegranate pericarp (peel) polyphenols (P) or pomegranate seed oil (Oil). Invasion across Matrigel by PC-3 human prostate cancer cells was measured following treatment with combinations of W, P and Oil such that the total gross weight of pomegranate extract was held constant. Expression of PLA2, associated with invasive potential, was measured in the PC-3 cells after treatment with the same dosage combinations as per invasion. Supra additive, complementary and synergistic effects were proven in all models by the Kruskal-Wallis non-parametric H test at p < 0.001 for the proliferation tests, p < 0.01 for invasion, and p < 0.05 for PLA2 expression. Proliferation effects were additionally evaluated with CompuSyn software median effect analysis and showed a concentration index CI < 1, confirming synergy. The results suggest vertical as well as the usual horizontal strategies for discovering pharmacological actives in plants. PMID- 15528978 TI - Phase I trial of orally administered CEP-701, a novel neurotrophin receptor linked tyrosine kinase inhibitor. AB - PURPOSE: A phase I clinical trial in patients with advanced carcinomas was conducted using the orally available neurotrophin receptor-linked tyrosine kinase receptor inhibitor, CEP-701. The objectives were to determine the maximum tolerated dose (MTD), dose-limiting toxicities, and pharmacokinetic profile of this orally administered agent. PATIENTS AND METHODS: A total of 30 patients were accrued to receive escalating BID doses of CEP-701 in cycles lasting 28 days. Between 3 and 6 patients were enrolled at each dose level. Once the MTD was determined, nine de novo patients were recruited to receive that level of drug. Pharmacokinetic studies were performed after the first dose, with additional sampling to assess intraindividual variability. RESULTS: The dosages ranged from 5 mg BID to 160 mg BID. While the criteria for MTD were not met at the dose levels administered, DLTs were observed at 80 and 120 mg BID. Treatment related adverse events, especially of the gastrointestinal system, made CEP-701 poorly tolerated at dosages above 40 mg BID. While CEP-701 did not produce an objective tumor response in any patient, 7 of the 30 patients received treatment for 3 months or more, including 3 who were on study with stable disease for more than 6 months. Orally administered CEP-701 was rapidly absorbed, with a mean t(max) between 1 and 3 hours. At higher dose levels, serum drug levels showed greater than dose-proportional increases by Day 28 versus Day 1. CONCLUSION: CEP-701 40 mg BID was well tolerated by patients with advanced malignancy and is the recommended dose level for planned phase II trials. Further study is necessary to determine the clinical efficacy of this novel new chemotherapeutic agent. PMID- 15528979 TI - Phase I pharmacokinetic studies evaluating single and multiple doses of oral GW572016, a dual EGFR-ErbB2 inhibitor, in healthy subjects. AB - GW572016 is a dual EGFR-ErbB2 inhibitor that has promise as an anticancer agent. Two phase I studies were conducted to determine the safety, tolerability and pharmacokinetics of single and multiple doses given to healthy subjects. The single dose study evaluated two groups of eight subjects in an ascending dose, 4 way cross-over, while the multiple dose study evaluated twenty-seven healthy volunteers in an ascending dose, double-blind, randomized, placebo-controlled, staggered parallel design. No serious adverse events were seen in either study. The most common adverse events for subjects receiving GW572016 were headache, diarrhea, rash, cold symptoms, gastrointestinal symptoms, and elevated LFTs, which were similar between treatment and placebo groups. Absorption of single doses of GW572016 was slightly delayed, with median t(lag) of 15 minutes (range 0 90 minutes) and achieved peak serum concentrations at a median of three hours (range 1.5-6 hours) post-dose. Serum concentrations after multiple doses of GW572016 demonstrated no significant accumulation at the 25 mg dose, and approximately 50% accumulation at the 100 mg and 175 mg doses, achieving steady state in six to seven days. A modest time-dependent increase in serum concentrations also was detected with multiple doses of GW572016. Single and multiple oral doses of GW572016 were well tolerated in healthy subjects, and resulted in dose-related systemic exposure of GW572016. PMID- 15528980 TI - Phase I study of Tomudex and Doxorubicin in patients with locally advanced, inoperable or metastatic cancer (IND.98). AB - BACKGROUND: The primary objective of this Phase I study was to determine the maximum tolerated dose (MTD) and recommended phase II dose for Tomudex and Doxorubicin when given in combination to patients with advanced metastatic cancer. The secondary objective was to assess the toxicity profile. PATIENTS AND METHODS: Starting doses were Tomudex 2.5 mg/m2 i.v. bolus day 1 and Doxorubicin 30 mg/m2 i.v. bolus day 1, repeated every 3 weeks. Doxorubicin was escalated in increments of 10 mg/m2 to 60 mg/m2, followed by escalation of Tomudex in increments of 0.5 mg/m2 to 3.5 mg/m2, on six dose levels. Twenty-five patients received 127 cycles of therapy, with at least 3 patients treated at each dose level. RESULTS: There was no dose limiting toxicity (DLT) observed in the first five dose levels. Three of six patients on dose level six had DLT. Further dose escalation was not warranted and this was declared the MTD. Grade 3 or 4 granulocytopenia was observed in 16/25 patients, with associated fever in 3/25 patients. Responses were seen in this study with one complete response (duration 12.8 months) and 3 partial responses (median duration 8 months) in 21 evaluable patients. Fourteen patients had stable disease (median duration 2.5 months). All 4 responding patients and 10 patients with stable disease had gastric cancer. CONCLUSIONS: The recommended phase-II dose for this combination in future studies is Tomudex 3 mg/m2 and Doxorubicin 60 mg/m2 given every 3 weeks. PMID- 15528981 TI - Phase I study of liposomal doxorubicin (Doxil) and cyclophosphamide in solid tumors. AB - INTRODUCTION: The purpose of this study was to determine the recommended phase II dose (RPTD) of the combination of cyclophosphamide and liposomal doxorubicin (Doxil). PATIENTS AND METHODS: Eligibility criteria included: a diagnosis of non hematologic cancer with no conventional effective therapy, normal renal, liver and bone marrow function, and ECOG performance status of 0-2. Both drugs were administered intravenously on day 1 of a 21-day cycle. The following doses (mg/m2) of Doxil and cyclophosphamide were tested: 30/500, 40/600, and 48/600. RESULTS: Thirty-seven patients with a median age of 62 years (male:female 21:16) received 119 cycles of Doxil and cyclophosphamide. Hand-foot syndrome and mucositis were the dose limiting toxicities for this combination. Other major toxicities included neutropenia. The side effects to the combination were cumulative. The RPTD of Doxil and cyclophosphamide was 48 mg/m2 and 600 mg/m2, respectively. Three patients with adenocarcinoma of the stomach, fibrosarcoma of the stomach and renal cell carcinoma showed objective antitumor responses. CONCLUSION: The toxicity profile of the combination of Doxil/cyclophosphamide differs significantly from that of non-liposomal doxorubicin plus cyclophosphamide. Major toxicities of the combination include the hand-foot syndrome, stomatitis and neutropenia. PMID- 15528982 TI - An NCIC CTG phase I/pharmacokinetic study of the matrix metalloproteinase and angiogenesis inhibitor BAY 12-9566 in combination with 5-fluorouracil/leucovorin. AB - BACKGROUND: This phase I study was performed to evaluate the safety, tolerability, and efficacy of the oral matrix metalloproteinase inhibitor BAY 12 9566 in combination with 5-fluorouracil/leucovorin in patients with advanced solid tumours, and to identify the maximum tolerated dose and the dose for use in future studies. PATIENTS AND METHODS: BAY 12-9566 and 5-fluorouracil/leucovorin were administered to 17 patients in 3 cohorts. Each patient served as his/her own control, with 5-fluorouracil being given alone on days 1-5 of cycle 1. In cohort 1, BAY 12-9566 at 800 mg p.o. b.i.d. was given with 350 mg/m2 5-fluorouracil/20 mg/m2 leucovorin x 5 days q28 days. In cohort 2, the BAY 12-9566 dose was reduced to 400 mg p.o. b.i.d., with the 5-fluorouracil/leucovorin doses remaining unchanged. Finally, in cohort 3, BAY 12-9566 400 mg bid was given with 5 fluorouracil 400 mg/m2/day. Patients were continued on therapy until unacceptable toxicity or tumour progression occurred. Pharmacokinetic analyses for both BAY 12 9566 and 5-fluorouracil were performed. RESULTS: The maximum tolerated dose was 400 mg p.o. b.i.d. BAY 12-9566 plus 5-fluorouracil/leucovorin at 400 mg/m2/day and 20 mg/m2/day, respectively. Thrombocytopenia necessitated a decrease of the dose of BAY 12-9566 by 50% from cohort 1 to cohort 2. Two dose-limiting toxicities occurred in cohort 3 consisting of neutropenic fever, and ileitis, causing severe diarrhea. Of 17 patients treated on study, 7 of 14 patients evaluable for response achieved stable disease. Pharmacokinetic analysis suggested there was no interaction between BAY 12-9566 and 5-fluorouracil. CONCLUSIONS: BAY 12-9566 400 mg bid and 5-fluorouracil 350 mg/m2 plus leucovorin 20 mg/m2 can be co-administered. Although there is some evidence of a clinical interaction, there is no apparent pharmacokinetic interaction. Future studies with these 2 types of agents administered in combination are warranted. PMID- 15528983 TI - Activity of gefitinib in advanced non-small-cell lung cancer with very poor performance status. AB - Advanced non-small-cell lung cancer (NSCLC) patients with poor performance status (PS) are less likely to respond to chemotherapy, or to have an improvement in survival, but more likely to experience toxicity. We retrospectively evaluated the efficacy and tolerability of gefitinib in patients with advanced NSCLC and very poor PS in Taiwan. Patients with stage IIIB, IV NSCLC with an Eastern Cooperative Oncology Group (ECOG) PS of 3-4 received oral gefitinib 250 mg once daily. Totally, 52 patients were included (25 men, 27 women). Forty-three patients (82.7%) were in a PS of 3. Tumor response rate was 25.0% (13/52). Tumor response rate to gefitinib was highest in chemonaive patients 38.1% (8/21) vs. failed 1 chemotherapy regimen 13.3% (2/15) vs. failed 2 or more chemotherapy regimens 18.8% (3/16), p = 0.015. The median overall survival was 2.5 months (response group 9.1 months, stable disease 3.1 months, and progressive group 0.8 month, p < 0.001). Adverse events, mainly skin reactions and diarrhea, were generally mild (grade 1 or 2) except paronychia and acne. Thus, gefitinib has clinically antitumor activity and good tolerability in Taiwan patients with advanced NSCLC and very poor performance status, with a higher response rate than that seen Europe or in European heritage Americans. Chemonaive patients responded better than patients with prior chemotherapy. Formal clinical trials are warranted to evaluate the role of gefitinib in this situation. PMID- 15528984 TI - Phase II study of oral bis (aceto) ammine dichloro (cyclohexamine) platinum (IV) (JM-216, BMS-182751) given daily x 5 in hormone refractory prostate cancer (HRPC). AB - JM-216 is an orally bioavailable platinum compound with activity against many tumor models. The objective of this study was to determine the safety profile and anti-tumor activity of JM-216 in patients with hormone refractory prostate cancer (HRPC) when given orally daily x 5 days. In this open label phase II study JM-216 was administered orally at the dose of 120 mg/m2/d for 5 days every 4 weeks. Patients continued on the therapy until evidence of disease progression or intolerable toxicity developed. Dose escalation and de-escalation were allowed according to patient's tolerance. Thirty-nine patients were enrolled onto the study and received a total of 155 courses (median 2, range 1-16) of JM-216. Dose delays (77% of courses) and dose reductions (31% of courses) were common and were mainly due to myelosupression. Treatment was discontinued in 5 patients due to treatment related toxicities. One patient developed myelodysplastic syndrome 11 months after the start of treatment. The most frequent grade III or higher adverse events included thrombocytopenia (54%), neutropenia (52%), anemia (24%) nausea (13%), vomiting (16%) and diarrhea (28%). PSA response was assessed in 32 patients, 10 (26%) had partial response, 14 (36%) had stable disease while PSA progression was seen in 8 (21%) patients. Of 20 (54%) patients with measurable disease two patients had a documented partial response. Although JM-216 had moderate activity in HRPC when given on daily basis for 5 days, it is associated with significant treatment related toxicities in this patient population. PMID- 15528985 TI - A phase II study of sabarubicin (MEN-10755) as second line therapy in patients with locally advanced or metastatic platinum/taxane resistant ovarian cancer. AB - BACKGROUND: Sabarubicin (MEN-10755) is a third generation anthracycline, with a remarkable antitumor activity in human tumor xenografts, including doxorubicin resistant tumors. Phase I studies have shown that myelosuppression is the main toxicity of sabarubicin, while its cumulative cardiotoxicity is mild. METHODS: The aim of the study was to evaluate the activity and safety profile of sabarubicin in patients with locally advanced or metastatic ovarian cancer failing 1st line platinum and/or taxane based chemotherapy, and relapsing earlier than 6 months after the last chemotherapy. Eligible patients received sabarubicin at the dose of 80 mg/m2 (dose level 0) every 3 weeks over 30 minutes. Dose was to be escalated to 90 mg/m2 (dose level +1) after the 1st cycle in case of grade 0-1 toxicity, while it was to be reduced to 60 mg/m2 (dose level -1) in case of toxicity. Response was assessed every 2 courses according to WHO criteria. Toxicity was graded according to Common Toxicity Criteria version 2.0. Gehan's design was used for sample size determination. RESULTS: Nineteen patients were enrolled and received 65 courses. One patient had a confirmed partial response, 9 patients had stable disease, 5 patients had disease progression, 3 patients were not evaluable for response, while one patient had an early progressive disease. The duration of response was 88 days. Mean time to disease progression was 125 days (range 56-188). Median survival was 62 days (range 36-202). Hematologic toxicity was moderate, since grade 3-4 neutropenia was observed in 25 out of 52 courses at 80 mg/m2, and grade 4 neutropenia occurred in one out of 12 courses at 90 mg/m2. Other grade 3-4 toxicities were: fatigue (five cases), nausea (two cases), stomatitis, general health deterioration, anorexia, vomiting, abdominal pain, hyponatremia (one case each). Cardiac toxicity was observed in the study; in fact, left ventricular ejection fraction (LVEF) fell below 50% in 2 patients, and 3 patients had a >15% decrease of LVEF from baseline, but there were no signs/symptoms of congestive heart failure. CONCLUSIONS: Sabarubicin showed limited activity in patients with resistant ovarian cancer. However, the observed data on disease stabilization, together with the drug's overall manageable toxicity profile, may prompt to its further investigation in advanced ovarian cancer. PMID- 15528986 TI - 15th Joint Meeting of the "Netherlands Society for Glycobiology", the "Studiengruppe Glykobiologie der Gesellschaft fur Biochemie und Molekularbiologie" and the "Groupe Lillois de Glycobiologie", November 29-30, 2004, Wageningen, The Netherlands. Abstracts. PMID- 15528987 TI - Thiamine attenuates hypoxia-induced cell death in cultured neonatal rat cardiomyocytes. AB - Previous studies have demonstrated that thiamine (vitamin B1) has a cytoprotective effect against ischemic damage to the heart, and that heat shock protein 70 (Hsp70) is capable of protecting cardiac cells from lethal ischemia/hypoxia. We show here that thiamine has a cytoprotective effect on cultured neonatal rat cardiomyocytes under hypoxic insult, and also protects the cardiomyocytes against hypoxia-induced apoptosis; caspase-3 activation, PARP cleavage and DNA fragmentation are all inhibited. Moreover, it increases the level of Hsp70 protein in the cardiomyocytes even under prolonged hypoxic stress and its effects on hypoxia-induced cardiac cell death are antagonized by an Hsp70 inhibitor. These results suggest that the cytoprotective effect of thiamine in cardiomyocytes under hypoxic stress is due to its ability to induce Hsp70. PMID- 15528988 TI - Frameshift deletion mechanisms in Egyptian Duchenne and Becker muscular dystrophy families. AB - Partial gene deletion is the major type of mutation leading to Duchenne muscular dystrophy (DMD) and its mild allelic form, Becker muscular dystrophy (BMD). Amplification of the genomic DNAs of 152 unrelated dystrophin patients using multiple primers detected 78 (51.3%) probands with deletion mutations. We predicted the translational reading frame for all the deletions in Egyptian dystrophin males. The frameshift rule was confirmed positively ranging for 50 to 67% of the cases depending on the type of disease. We discuss ways of accounting for some exceptions from the frameshift hypothesis in the central and proximal regions. These explanations may help in developing procedures for reducing the severity of dystrophin phenotypes to restore the correct frame by disrupting the translational fidelity. Great efforts have been put into the development of effective 'gene correction' procedures via such intrinsic mechanisms. In addition, we mapped regional difference in deletion mutation frequencies within the DMD gene locus between the different Egyptian governorates. There were no double deletions in the Egyptian dystrophin males. PMID- 15528989 TI - Cloning and functional analysis of a cDNA encoding Ginkgo biloba farnesyl diphosphate synthase. AB - Farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2. 5.1.10) catalyzes the synthesis of farnesyl diphosphate, and provides precursor for biosynthesis of sesquiterpene and isoprenoids containing more than 15 isoprene units in Ginkgo biloba. Here we report the cloning, characterization and functional analysis of a new cDNA encoding FPS from G. biloba. The full-length cDNA (designated GbFPS) had 1731 bp with an open reading frame of 1170 bp encoding a polypeptide of 390 amino acids. The deduced GbFPS was similar to other known FPSs and contained all the conserved regions of trans-prenyl chain-elongating enzymes. Structural modeling showed that GbFPS had the typical structure of FPS, the most prominent feature of which is the arrangement of 13 core helices around a large central cavity. Southern blot analysis revealed a small FPS gene family in G. biloba. Expression analysis indicated that GbFPS expression was high in roots and leaves, and low in stems. Functional complementation of GbFPS in an FPS-deficient strain confirmed that GbFPS mediates farnesyl diphosphate biosynthesis. PMID- 15528990 TI - Regulatory elements involved in transcription of the human NeuAcalpha2,3Galbeta1,3GalNAcalpha2,6-sialyltransferase (hST6GalNAc IV) gene. AB - We previously cloned and characterized the promoter region of the human NeuAcalpha2,3Galbeta1,3GalNAcalpha2,6-sialyltransferase (hST6GalNAc IV) gene [Kim et al. (2003)]. In the present study, we identified a region of 294 bp upstream of exon 1 of the gene that produced maximal transcriptional activity in human Jurkat T cells. Site-directed mutagenesis and transient transfection assays demonstrated that Sp1 and MZF1 elements in this region were required for the promoter activity. Further analysis by electrophoretic mobility shift assays using specific competitors and antibody revealed that Sp1 and MZF1 nuclear proteins interacted with these elements. These results indicate that Sp1 and MZF1 are involved in the transcriptional regulation of the hST6GalNAc IV gene in Jurkat T cells. PMID- 15528991 TI - Inhibitory effects of glycoprotein-120 (G-120) from Ulmus davidiana Nakai on cell growth and activation of matrix metalloproteinases. AB - Excessive breakdown of extracellular matrix by metalloproteinases (MMPs) occurs in many pathological conditions. Consequently, methods for inhibiting MMP activity have therapeutic potential. In this study, we investigated the effect of G-120, a 120 kDa glycoprotein purified from the Oriental herbal plant, Ulmus davidiana Nakai (UDN), on the activity and production of several MMPs by evaluating its growth inhibitory effect on NIH 3T3 cells. Tritium uptake assays showed that proliferation of NIH 3T3 cells was strongly suppressed, and G-120 mediated inhibition of DNA synthesis proved to involve a cytostatic, rather than a cytotoxic, effect, as shown by cytotoxicity and apoptosis assays. More importantly, G-120 strongly reduced the gelatinolytic and collagenase activities of MMP proteins, as well as expression of MMP-2 and MMP-9. Electrophoretic mobility shift assays revealed that it suppressed the DNA binding activity of NF kappaB. Collectively, our observations show that G-120 strongly inhibits the activation of MMPs and NF-kappaB. PMID- 15528992 TI - Expression of 17beta-hydroxysteroid dehydrogenease in testis of the ascidian Ciona intestinalis [corrected]. AB - Ascidians have been employed as model organisms in investigating spermatogenesis. 17beta-hydroxysteroid dehydrogenase (HSD) is a steroidogenic enzyme essential for invertebrate spermatogenesis. A homologue of HSD was found in the EST database of Ciona intestinalis and cloned. Sequence analysis showed significant homology to zebra fish, sea urchin and human 17beta-HSD. The gene has an open reading frame (ORF) of 918 nucleotides coding for a polypeptide of 306 amino acids and a calculated mass of 35-kDa. Immunoblotting with an antibody raised against HSD recognized a 35-kDa protein purified from the C. intestinalis testis. The HSD protein was localized in steroidogenic cells in the Ciona testis. These results suggest that C. intestinalis 17beta-HSD is equivalent to the enzyme of vertebrate Leydig cells and that 17beta-HSD could be a phylogenetic marker for organisms producing steroids. PMID- 15528993 TI - Trichomonas vaginalis inhibits proinflammatory cytokine production in macrophages by suppressing NF-kappaB activation. AB - Activation of NF-kappaB leads to the production of proinflammatory cytokines such as IL-12 and TNF-alpha that are involved in innate and adaptive immunity. We determined whether T. vaginalis-induced inflammatory response in macrophages associated with NF-kappaB. T. vaginalis adhesion led to transient NF-kappaB activation at 6 h but activation declined dramatically by 8 h. Super-shift assays showed that the gel-shifted complexes consisted of p65 (Rel A) and p50 (NF kappaB1). NF-kappaB activation was accompanied by IkappaB-alpha degradation, and was inhibited by blocking T. vaginalis adhesion, indicating that the early NF kappaB activation by T. vaginalis depends on IkappaB-alpha degradation. Quantitative real-time RT-PCR analyses revealed that the expression of TNF-alpha and IL-12 mRNA in T. vaginalis-adhesive cells was rapidly suppressed in comparison with LPS stimulation. We also observed that the parasite inhibited the nuclear translocation of NF-kappaB at 8 h, and diminished IL-12 and TNF-alpha production in response to LPS. In addition, inhibition of IkappaB-alpha degradation by MG-132 resulted in apoptosis. These results demonstrate that effects of T. vaginalis on NF-kappaB regulation are critical for cytokine production and the survival of macrophages. We suggest that there exist inhibitory mechanisms induced by T. vaginalis to evade host immunity. PMID- 15528994 TI - CD99 costimulation up-regulates T cell receptor-mediated activation of JNK and AP 1. AB - Although CD28 is the principal T cell costimulatory molecule for the T cell receptor, a number of other cell surface proteins have costimulatory functions and perform specific roles in different contexts. Here we analyzed the mechanism of CD99 costimulation of the T cell receptor. Cooperation of CD99 engagement with suboptimal TCR/CD3 signals resulted in greatly enhanced CD4+ T cell proliferation. CD99 costimulation also led to elevated expression of CD25 and GM1 on the CD4+ T cell surface within 3 days. In Jurkat TAg cells, CD99 costimulation led to increased apoptosis compared to stimulation with CD3 or CD99 alone. CD99 costimulation also augmented activation of MAP kinases, especially of JNK, and increased AP-1 activation was also observed using a luciferase reporter assay. These results show that CD99 has a costimulatory function for T cells and acts by a mechanism distinct from CD28. PMID- 15528995 TI - Nucleotide sequences of a Korean isolate of apple stem grooving virus associated with black necrotic leaf spot disease on pear (Pyrus pyrifolia). AB - Pear black necrotic leaf spot (PBNLS) is a disease of pears caused by capillovirus-like particles, which can be observed under the electron microscope. The disease was analyzed by Western blot analysis with antisera raised against apple stem grooving virus (ASGV) coat protein. cDNAs covering the entire genome were synthesized by RT-PCR and RACE using RNA isolated from Chenopodium quinoa infected with sap extracted from pear leaves carrying black necrotic spot disease. The complete genome sequence of the putative pear virus, 6497 nucleotides in length excluding the poly (A) tail, was determined and analyzed. It contains two overlapping open reading frames (ORFs). ORF1, spans from nucleotide position 37 to 6354, producing a putative protein of 241 kDa. ORF2, which is in a different reading frame within ORF1, begins at nucleotide 4788 and terminates at 5750, and produces a putative protein of 36 kDa. The 241 kDa protein contains sequences related to the NTP-binding motifs of helicases and RNA dependent RNA polymerases. The 36-kDa protein contains the consensus sequence GDSG found in the active sites of several cellular and viral serine proteases. Morphological and serological analysis, and sequence comparison between the putative pear virus, ASGV, citrus tatter leaf virus and cherry virus A of the capillovirus suggest that PBNLS may be caused by a Korean isolate of ASGV. PMID- 15528996 TI - IKKgamma inhibits activation of NF-kappaB by NIK. AB - IKKgamma is a component of the IKK complex, which regulates NF-kappaB activity. To investigate the role of IKKgamma, we expressed wild type IKKgamma containing 412 amino acids, and deletion mutants containing residues 1-312 and 101-412, using murine IKKgamma cDNA. In a co-transfection assay with a CAT reporter plasmid, NIK activated NF-kappaB-dependent gene expression approximately two fold and this expression was inhibited by co-transfection of a wild type IKKgamma expression plasmid. In binding assays IKKgamma inhibited the association of IkappaBalpha with IKKbeta and the subsequent phosphorylation of IkappaBalpha that is activated by NIK. Inhibition by IKKgamma also occurred in an assay with a dominant negative mutant of NIK but not with a C-terminal deletion mutant of IKKgamma, indicating that the C-terminal 100 amino acids of IKKgamma are important for negative regulation of NF-kappaB activation. In addition, the interaction of IKKbeta with IKKgamma was inhibited by co-transfection with a NIK expression plasmid. Our results suggest that overexpression of IKKgamma inhibits activation of NF-kappaB by NIK by competing with NIK for interaction with IKKbeta. PMID- 15528997 TI - Pathogenesis-related gene expression by specific calmodulin isoforms is dependent on NIM1, a key regulator of systemic acquired resistance. AB - Plants produce numerous calmodulin isoforms that exhibit differential gene expression patterns and sense different Ca2+ signals. This diversity results in different physiological responses to particular stimuli. Gm-CaM-4 and -5 are two divergent calmodulin isoforms from the soybean (Glycine max) that have been reported to be involved in plant disease resistance. However, little is known about the pathway by which these specific isoforms transduce the defense signal and up-regulate pathogenesis-related (PR) genes. Here we report that overexpression of GmCaM-4/-5 induces constitutive PR gene expression and enhances disease resistance in wild-type Arabidopsis, but not in the nim1 mutant of Arabidopsis. GmCaM-4/-5 also appear to activate trans-acting elements that bind to cis-acting elements in the Arabidopsis PR-1 promoter. Thus up-regulation of PR genes by these GmCaM isoforms is dependent on NIM1 (Non immunity 1) and unknown transcription factors. PMID- 15528998 TI - Cysteine-321 of human brain GABA transaminase is involved in intersubunit cross linking. AB - Gamma-aminobutyrate transaminase (GABA-T), a key homodimeric enzyme of the GABA shunt, converts the major inhibitory neurotransmitter GABA to succinic semialdehyde. We previously overexpressed, purified and characterized human brain GABA-T. To identify the structural and functional roles of the cysteinyl residue at position 321, we constructed various GABA-T mutants by site-directed mutagenesis. The purified wild type GABA-T enzyme was enzymatically active, whereas the mutant enzymes were inactive. Reaction of 1.5 sulfhydryl groups per wild type dimer with 5,5 cent-dithiobis-2-nitrobenzoic acid (DTNB) produced about 95% loss of activity. No reactive -SH groups were detected in the mutant enzymes. Wild type GABA-T, but not the mutants, existed as an oligomeric species of Mr = 100,000 that was dissociable by 2-mercaptoethanol. These results suggest that the Cys321 residue is essential for the catalytic function of GABA-T, and that it is involved in the formation of a disulfide link between two monomers of human brain GABA-T. PMID- 15528999 TI - Deregulation of catalase, not MnSOD, is associated with necrotic death of p53 defective DF-1 cells under antimycin A-induced oxidative stress. AB - One of distinct genetic alterations in spontaneously immortalized DF-1 cells was found to be dysfunction of p53 and E2F-1 as well as altered antioxidant gene expression (upregulation of MnSOD and downregulation of catalase). We have characterized the cellular responses of primary and immortal DF-1 cells to oxidative stress and found that DF-1 cells were more sensitive to oxidative stress than their primary counterparts when treated with antimycin A. The increased DF-1 cell death by oxidative stress was accompanied by an increase in the levels of intracellular superoxide anions and hydrogen peroxide. The cell death in DF-1 cells by antimycin A showed none of the hallmarks of apoptosis, but displayed a significantly increased necrotic cell population. Anti-apoptotic Bcl 2 failed to inhibit oxidative-induced necrotic cell death in the DF-1 cells. However, this necrotic cell death was significantly decreased by treatment with hydrogen peroxide scavengers such as sodium pyruvate and N-acetyl-cysteine. Interestingly, overexpression of human catalase in DF-1 cells endowed cells resistant to the oxidative stress by antimycin A treatment, although the downregulation of MnSOD by an antisense strategy showed no evident change in the cytotoxic effect caused by antimycin A. Taken together, the present study might provide new therapeutic approach for tumor cells having the loss of p53 function and the altered antioxidant functions. PMID- 15529000 TI - Precocious expression of Drosophila Rbp9 inhibits ovarian germ cell proliferation. AB - We previously reported that Rbp9 is expressed in the germarium region as soon as germ cells complete mitosis and form 16 interconnected cystocytes. Mutation of Rbp9 caused over-proliferation of cystocytes and generated an ovarian tumor phenotype. This led us to conclude that Rbp9 either inhibits cell proliferation or is required for cell differentiation. To examine the role of Rbp9, we over expressed it ectopically in germline stem cells and early stage cystocytes that had not yet formed 16 cell clusters. The egg chambers of the newly eclosed transgenic flies looked normal. However, on day 12, most of the developing egg chambers had disappeared and only the germarium region remained. Staining of the Rbp9 over-expressing transgenic ovaries with mitosis markers revealed almost no mitotic divisions in the transgenic germaria. Further examination of these cystocytes with HTS [hu-li tai shao protein], which labels the fusome structure, revealed that the cells in the germarium region consisted of stem cells and cystocytes that had not yet completed four mitotic divisions. These observations suggest that precocious expression of Rbp9 inhibits cystocyte proliferation. Therefore the precise onset of Rbp9 expression in germarium region 2a is critical for inhibiting their further proliferation and achieving their correct differentiation. PMID- 15529001 TI - A monoclonal antibody against the paraneoplastic pemphigus (PNP) antigen, envoplakin: cDNA sequences encoding the variable regions of heavy and light chains. AB - Paraneoplastic pemphigus (PNP) is an acquired autoimmune disorder characterized by severe mucosal erosion, and polymorphous cutaneous lesions associated with neoplasia. PNP patients have circulating autoantibodies that bind to stratified and nonstratified epithelia. Previously, we showed that envoplakin was a component of the antigen complex recognized by PNP sera. In the present study we generated a monoclonal antibody, EVP-YS, against human envoplakin. The antibody bound to keratinocyte cell surfaces and reacted with the 210-kDa PNP antigen, confirming its specificity for envoplakin. The variable regions of the heavy (H) and light (L) chain genes were cloned from the hybridoma and shown to belong to mouse H chain subgroup III and kappa light chain subgroup V, respectively. The L chain of EVP-YS was 98% identical to the k chains of some autoantibodies and anti nucleic acid antibodies, and had an identical amino acid sequence in all three complementary determining regions, suggesting that the H chains determine the specificity of the EVP-YS-envoplakin interaction. The EVP-YS antibody can be used to evaluate the sensitivity and specificity of clinical, histological, and immunological criteria for diagnosing PNP. PMID- 15529002 TI - Transcriptional regulation of glutathione synthetase in the fission yeast Schizosaccharomyces pombe. AB - Glutathione (GSH), an important antioxidant involved in the stress response, is synthesized in two sequential reactions involving glutamylcysteine synthetase (GCS), followed by glutathione synthetase (GS). Expression of the unique GS gene in the fission yeast Schizosaccharomyces pombe was previously found to be regulated by nitric oxide and by L-buthionine-(S,R)-sulfoximine (BSO), a specific inhibitor of GCS. In this work, expression of S. pombe GS gene is shown to be induced by menadione (MD), which generates superoxide. The responsible DNA sequence between -365 and -234 bp from the translation start site, was convinced using five GS-lacZ fusion plasmids. Expression of GS gene is also induced by low glucose, fructose and disaccharides, apparently dependent on Pap1 protein; GS mRNA increases in low concentrations of glucose in wild type S. pombe but not in Pap1-negative cells. Although nonfermentable carbon sources such as acetate and ethanol stimulate expression of GS gene, they also arrest the growth of the yeast cells. These results indicate that the biosynthesis of glutathione is regulated by superoxide radicals and carbon source limitation. PMID- 15529003 TI - Repair of mitomycin C cross-linked DNA in mammalian cells measured by a host cell reactivation assay. AB - DNA repair capacity in a cell could be detected by a host-cell reactivation assay (HCR). Since relation between DNA repair and genetic susceptibility to cancer remains unclear, it is necessary to identify DNA repair defects in human cancer cells. To assess DNA repair for breast cancer susceptibility, we developed a modified HCR assay using a plasmid containing a firefly luciferase gene damaged by mitomycin C (MMC), which forms interstrand cross-link (ICL) adducts. In particular, interstrand cross-link is thought to induce strand breaks being repaired by homologous recombination. The MMC-ICLs were verified by electrophoresis. Damaged plasmids were transfected into apparently normal human lymphocytes and NER-deficient XP cell lines and the DNA repair capacity of the cells were measured by quantifying the activity of the firefly luciferase. MMC lesion was repaired as much as UV adducts in normal lymphocytes and the XPC cells. However, the XPA cells have a lower repair capacity for MMC lesion than the XPC cell, indicating that the XPA protein may be involved in initial damage recognition of MMC-ICL adducts. Since several repair pathways including NER and recombination participate in MMC-ICL removal, this host cell reactivation assay using MMC-ICLs can be used in exploring DNA repair defects in human cancer cells. PMID- 15529004 TI - Cloning and circadian expression of rat Cry1. AB - In mammals, two types of cryptochrome are involved in the regulation of the circadian rhythm. We previously characterized rat Cry2 and its expression in brain tissue [Eun et al. (2001)]. We report here the cloning of another cryptochrome gene, Cry1, from rat brain by reverse-transcription coupled to polymerase chain reaction (RT-PCR), together with rapid-amplification of cDNA ends (RACE). The cloned Cry1 cDNA consists of 2557 nucleotides and has a single open-reading frame encoding a protein of 588 amino acids with start and stop codons. The deduced amino acid sequence was 70% identical with that of rat Cry2. It also showed 95% identity with mouse and human Cry1 but relatively low identity of 82% with that of zebrafish. Circadian expression of rat Cry1 and Cry2 was examined in the suprachiasma nucleus (SCN) and eye by real-time PCR. Expression of Cry1 and Cry2 mRNA in the SCN displayed a circadian rhythm with a peak at the day/night transition, and there was a slightly different circadian pattern of expression of Cry1 and Cry2 in the eye. PMID- 15529005 TI - Establishment of life-span extended bovine fibroblast cells carrying the characterization of primary cells. AB - Although primary bovine embryonic fibroblast (BEF) cells have previously been used as nucleus-donors for nuclear transfer (NT), it has now been proposed to use BEF cells to generate cloned cows that were genetically modified by transgenic or a knock-out system. A major limitation to gene targeting somatic cells, however, is the overall life-span of the cell. In this study, we first examined in vitro life-span of primary BEF cells. Primary BEF cells were found to be replicative senescent at passage 10th-12th, similar to primary murine embryonic fibroblast cells. To overcome this short in vitro life-span, we have optimized culture conditions to extend the life-span and determined growth characteristics of BEF cell lines. Two life-span extended BEF cell lines (designated CGFR -BO-1 and CGFR BO-2) were shown to grow much faster than their parental primary counterparts. Both cell lines did not display any potential for abnormal growth such as foci formations in either soft-agar or confluent culture condition. In cloning experiments using these cell lines as a nuclear donor, the reconstructed karyoblasts underwent apoptosis, reprogramming and development in the blastocyst stage, at a similar frequency to those observed with parental as well as adult primary fibroblasts. Furthermore, these cell lines targeted with green fluorescence protein (GFP) were successfully transduced, selected and reprogrammed by NT to develop into a blastocyst stage with GFP expression. Our results suggested methods to extend life-span of donor cells with tremendous implications for the genetic engineering of bovine fibroblast cells. PMID- 15529007 TI - Modeling cerebral arteriovenous lactate kinetics after intravenous lactate infusion in the rat. AB - Venous-arterial lactate differences across the brain during lactate infusion in rats were studied, and the fate of lactate was described with a mathematical model that includes both cerebral and extracerebral kinetics. Ultrafiltration was used to sample continuously and simultaneously arterial and venous blood. Subsequent application of flow injection analysis and biosensors allowed the measurement of glucose and lactate concentrations every minute. Because of the high temporal resolution, arteriovenous lactate kinetics could be modeled in individual experiments. The existence of both a cerebral lactate sink and a lactate exchangeable compartment, representing approximately 24% of brain volume, was thus modeled. PMID- 15529008 TI - Identification of hemodynamic compromise by cerebrovascular reserve and oxygen extraction fraction in occlusive vascular disease. AB - Cerebrovascular reserve (CVR) and oxygen extraction fraction (OEF) are used to identify hemodynamic compromise in symptomatic patients with carotid occlusive vascular disease, but evidence suggests that they are not equivalent. The authors studied the relationship between CVR and OEF to evaluate their equivalence and stages of hemodynamic compromise. Symptomatic patients (N = 12) with carotid occlusion were studied by stable xenon-computed tomography CBF after intravenous acetazolamide administration for CVR, followed within 24 hours by positron emission tomography (PET) for OEF. Middle cerebral artery territories were analyzed by hemisphere and level. Hemispheric subcortical white matter infarctions were graded with magnetic resonance imaging. Both hemispheric and level analysis of CVR and OEF showed a significant (P = 0.001), negative linear relationship [CVR (%) = -1.5 (OEF) + 83.4, (r = -0.57, P = 0.001, n = 24]. However, 37.5% of the hemispheres showed compromised CVR but normal OEF and were associated (P = 0.019) with subcortical white matter infarction. CMRO2 was elevated in stage II hemodynamic compromise (CVR < 10%, OEF > 50%). CVR and OEF showed a significant negative linear relationship in stage II hemodynamic compromise but revealed hemispheres in hemodynamic compromise by CVR but normal OEF that were associated with subcortical white matter infarction. PMID- 15529009 TI - Secondary energy failure after cerebral hypoxia-ischemia in the immature rat. AB - A delayed or secondary energy failure occurs during recovery from perinatal cerebral hypoxia-ischemia. The question remains as to whether the energy failure causes or accentuates the ultimate brain damage or is a consequence of cell death. To resolve the issue, 7-day postnatal rats underwent unilateral common carotid artery occlusion followed thereafter by systemic hypoxia with 8% oxygen for 2.5 hours. During recovery, the brains were quick frozen and individually processed for histology and the measurements of 1) high-energy phosphate reserves and 2) neuronal (MAP-2, SNAP-25) and glial (GFAP) proteins. Phosphocreatine (PCr) and ATP, initially depleted during hypoxia-ischemia, were partially restored during the first 18 hours of recovery, with secondary depletions at 24 and 48 hours. During the initial recovery phase (6 to 18 hours), there was a significant correlation between PCr and the histology score (0 to 3), but not for ATP. During the late recovery phase, there was a highly significant correlation between all measured metabolites and the damage score. Significant correlation also exhibited between the neuronal protein markers, MAP-2 and SNAP-25, and PCr as well as the sum of PCr and Cr at both phases of recovery. No correlation existed between the high-energy reserves and the glial protein marker, GFAP. The close correspondence of PCr to histologic brain damage and the loss of MAP-2 and SNAP-25 during both the early and late recovery intervals suggest evolving cellular destruction as the primary event, which precedes and leads to the secondary energy failure. PMID- 15529010 TI - The AMPA receptor potentiator LY404187 increases cerebral glucose utilization and c-fos expression in the rat. AB - AMPA receptor potentiators enhance AMPA receptor-mediated glutamatergic neurotransmission and may have therapeutic potential as cognitive enhancers or antidepressants. The anatomical basis for the action of AMPA receptor potentiators is unknown. The aim of this study was to determine the effects of the biarylpropylsulfonamide AMPA receptor potentiator, LY404187 (0.05 to 5 mg/kg subcutaneously), upon cerebral glucose utilization and c-fos expression using 14C 2-deoxglucose autoradiography and c-fos immunocytochemistry. LY404187 (0.5 mg/kg) produced significant elevations in glucose utilization in 28 of the 52 anatomical regions analyzed, which included rostral neocortical areas and the hippocampus, as well the dorsal raphe nucleus, lateral habenula, and locus coeruleus. No significant decreases in glucose utilization were observed in any region after LY404187 administration. The increases in glucose utilization with LY404187 (0.5 mg/kg) were blocked by pretreatment with the AMPA receptor antagonist LY293558 (25 mg/kg), indicating that LY404187 acts through AMPA receptor-mediated mechanisms. LY404187 (0.5 mg/kg) also produced increases in c-fos immunoreactivity in the cortex, locus coeruleus, and the dorsal raphe nucleus. These studies demonstrate neuronal activation in key brain areas that are associated with memory processes and thus provide an anatomical basis for the cognitive enhancing effects of AMPA receptor potentiators. PMID- 15529011 TI - The chemokine fractalkine in patients with severe traumatic brain injury and a mouse model of closed head injury. AB - The potential role of the chemokine Fractalkine (CX3CL1) in the pathophysiology of traumatic brain injury (TBI) was investigated in patients with head trauma and in mice after experimental cortical contusion. In control individuals, soluble (s)Fractalkine was present at low concentrations in cerebrospinal fluid (CSF) (12.6 to 57.3 pg/mL) but at much higher levels in serum (21,288 to 74,548 pg/mL). Elevation of sFractalkine in CSF of TBI patients was observed during the whole study period (means: 29.92 to 535.33 pg/mL), whereas serum levels remained within normal ranges (means: 3,100 to 59,159 pg/mL). Based on these differences, a possible passage of sFractalkine from blood to CSF was supported by the strong correlation between blood-brain barrier dysfunction (according to the CSF-/serum albumin quotient) and sFractalkine concentrations in CSF (R = 0.706; P < 0.01). In the brain of mice subjected to closed head injury, neither Fractalkine protein nor mRNA were found to be augmented; however, Fractalkine receptor (CX3CR1) mRNA steadily increased peaking at 1 week postinjury (P < 0.05, one-way analysis of variance). This possibly implies the receptor to be the key factor determining the action of constitutively expressed Fractalkine. Altogether, these data suggest that the Fractalkine-CX3CR1 protein system may be involved in the inflammatory response to TBI, particularly for the accumulation of leukocytes in the injured parenchyma. PMID- 15529012 TI - Caspase inhibitor z-DEVD-fmk attenuates calpain and necrotic cell death in vitro and after traumatic brain injury. AB - In studies designed to evaluate the therapeutic window for treatment of traumatic brain injury, the caspase 3 inhibitor z-DEVD-fmk improved neurologic function and reduced lesion volumes when administered at 1 but not at 4, 8, or 24 hours after injury. Moreover, neither caspase 3 nor PARP, a caspase 3 substrate, were cleaved in injured, untreated cortex from 1 to 72 hours after injury. Few cortical neurons expressed active caspase 3 or were TUNEL positive from 6 to 24 hours after injury, and TUNEL staining was primarily Type I (necrotic). Nissl staining revealed extensive neuronal necrosis in the injured cortex from 6 to 24 hours after impact. Considered together, these data suggested that z-DEVD-fmk may reduce neuronal necrosis, so we used an in vitro model of necrotic cell death induced by maitotoxin to test this further and explore the potential mechanism(s) involved. Z-DEVD-fmk (1 nM-100 microM) significantly attenuated maitotoxin induced neuronal cell death and markedly reduced expression of the 145 kD calpain mediated alpha-spectrin breakdown product after maitotoxin injury. Neither the 120 kD caspase-mediated alpha-spectrin cleavage product nor cathepsin B were expressed after maitotoxin injury. In a cell free assay, z-DEVD-fmk reduced hydrolysis of casein by purified calpain I. Finally, z-DEVD-fmk reduced expression of the 145 kD calpain-mediated alpha-spectrin cleavage fragment after traumatic brain injury in vivo. These data suggest that neuroprotection by z-DEVD fmk may, in part, reflect inhibition of calpain-related necrotic cell death. PMID- 15529013 TI - Mmp-9 deficiency enhances collagenase-induced intracerebral hemorrhage and brain injury in mutant mice. AB - Matrix metalloproteinase-9 (MMP-9) participates in the disregulation of blood brain barrier during hemorrhagic transformation, and exacerbates brain injury after cerebral ischemia. However, the consequences of long-term inhibition or deficiency of MMP-9 activity (which might affect normal collagen or matrix homeostasis) remains to be determined. The authors investigated how MMP-9 gene deficiency enhances hemorrhage and increases mortality and neurologic deficits in a collagenase-induced intracerebral hemorrhage (ICH) model in MMP-9-knockout mice. MMP-9-knockout and corresponding wild-type mice at 20 to 35 weeks were used to model an aged population (because advanced age is a significant risk factor in human ICH). Collagenase VII-S (0.5 microL, 0.075 U) was injected into the right basal ganglia in mice and mortality, neurologic deficits, brain edema, and hemorrhage size measured. In addition, MMP-9 activity, brain collagen content, blood coagulation, cerebral arterial structure, and expressions of several MMPs were examined. Increased hemorrhage and brain edema that correlated with higher mortality and neurologic deficits were found in MMP-9-knockout mice. No apparent structural changes were observed in cerebral arteries, even though brain collagen content was reduced in MMP-9-knockout mice. MMP-9-knockout mice did exhibit an enhanced expression of MMP-2 and MMP-3 in response to ICH. The results indicate that a deficiency of MMP-9 gene in mutant mice increases collagenase-induced hemorrhage and the resulting brain injury. The intriguing relationship between MMP-9 deficiency and collagenase-induced ICH may reflect the reduction in collagen content and an enhanced expression of MMP-2 and MMP-3. PMID- 15529014 TI - Increased severity of cerebral ischemic injury in vascular endothelial growth factor-B-deficient mice. AB - Vascular endothelial growth factor-B (VegfB) is an angiogenic protein related to VegfA, although it acts on a different set of tyrosine kinase receptors. Like VegfA, VegfB is expressed in the brain and is induced at sites of brain injury. VegfA has neuroprotective and angiogenic effects, but VegfA-knockout mice die in utero, so the effect of endogenous VegfA signaling in neuropathologic states, such as cerebral ischemia, cannot be tested directly. In contrast, VegfB-knockout mice survive to adulthood with little abnormality in the absence of pathologic stresses. To determine if VegfB regulates the severity of cerebral ischemia, the middle cerebral artery was occluded in VegfB-knockout, heterozygous, and wild type mice, and the volume of the resulting cerebral infarcts and associated impairment of neurologic function were measured. Infarct volume was increased by approximately 40% and neurologic impairment was more severe in VegfB-knockout mice, implying that endogenous VegfB acts to protect the brain from ischemic injury. VegfB also protected cultured cerebral cortical neurons from hypoxic injury, suggesting that its protective action is mediated at least in part through a direct effect on neurons. PMID- 15529015 TI - 2,7-Bis-(4-amidinobenzylidene)-cycloheptan-1-one dihydrochloride, tPA stop, prevents tPA-enhanced excitotoxicity both in vitro and in vivo. AB - Tissue-type plasminogen activator (tPA) is available for the treatment of thromboembolic stroke in humans. However, adverse effects of tPA have been observed in animal models of ischemic brain injuries. In the present study, we have used a synthetic tPA inhibitor, named 2,7-bis-(4-amidino-benzylidene) cycloheptan-1-one dihydrochloride (tPA stop), to investigate the role of endogenous tPA in the cerebral parenchyma. In mouse cortical cell cultures, we observed that although tPA stop reduced N-methyl-D-aspartic acid (NMDA)-mediated excitotoxic neuronal death, it failed to modulate alpha-amino-2,3-dihydro-5 methyl-3-oxo-4-isoxazole propanoic acid or kainate-mediated necrosis. In addition, we found that tPA stop could prevent the deleterious effects of both endogenous and exogenous tPA during NMDA exposure. At the functional level, tPA stop was found to prevent tPA-dependent potentiation of NMDA receptor-evoked calcium influx. The relevance of those findings was strengthened by the observation of a massive reduction of NMDA-induced excitotoxic lesion in rats when tPA stop was co-injected. Altogether, these data demonstrate that the blockade of the endogenous proteolytic activity of tPA in the cerebral parenchyma could be a powerful neuroprotective strategy raised against brain pathologies associated with excitotoxicity. PMID- 15529016 TI - Effects of combined estrogen and progesterone on brain infarction in reproductively senescent female rats. AB - Recent data from the Women's Health Initiative have highlighted many fundamental issues about the utility and safety of long-term estrogen use in women. Current hormone replacement therapy for postmenopausal women incorporates progestin with estrogen, but it is uncertain if combined therapy provides major cerebrovascular risks or benefits to these women. No experimental animal stroke studies have examined combined hormone administration. The authors tested the hypothesis that combined hormone treatment reduces ischemic injury in middle-aged female rat brain. Reproductively senescent female rats underwent 2-hour middle cerebral artery occlusion (MCAO) followed by 22 hours reperfusion. Estrogen implants were placed subcutaneously at least 7 days before MCAO, and progesterone intraperitoneal injections were given 30 minutes before MCAO, at initiation, and at 6 hours of reperfusion. Rats received no hormone, a 25-microg estrogen implant, a 25-microg estrogen implant plus 5 mg/kg intraperitoneal progesterone, or 5 mg/kg intraperitoneal progesterone. Cortical, caudoputamen, and total infarct volumes were assessed by 2,3,5-triphenyltetrazolium chloride staining and digital image analysis at 22 hours reperfusion. Cortical and total infarct volumes, except in the acute progesterone-treated group, were significantly attenuated in all estrogen-alone and combined hormone-treated groups. There were no significant differences in caudoputamen infarct volumes in all hormone-treated groups as compared with untreated rats. These data have potential clinical implications relative to stroke for postmenopausal women taking combined hormone replacement therapy. PMID- 15529017 TI - Induction of tolerance to focal ischemia in rat brain: dissociation between cortical lesioning and spreading depression. AB - Cortical application of KCl has previously been shown to induce tolerance to a subsequent episode of cerebral ischemia. KCl triggers recurrent episodes of cortical spreading depression and produces a small lesion at the cortical application site. To determine whether a cortical lesion alone is sufficient to induce tolerance to ischemia, the authors used 5-mol/L NaCl to precondition rat brain 3 days before permanent occlusion of the middle cerebral artery. NaCl produced a small lesion at the application site without evoking cortical spreading depression. Preconditioning with 5-mol/L NaCl significantly attenuated the decrease in CBF after middle cerebral artery occlusion and reduced the volume of cortical infarction by 35%. The results show that a small cortical lesion, by itself, is sufficient to induce tolerance to ischemia. PMID- 15529018 TI - Pronounced hypoperfusion during spreading depression in mouse cortex. AB - We studied unique cerebral blood flow (CBF) responses to cortical spreading depression in mice using a novel two-dimensional CBF imaging technique, laser speckle flowmetry. Cortical spreading depression caused a triphasic CBF response in both rat and mouse cortex. In rats, mild initial hypoperfusion (approximately 75% of baseline) was followed by a transient hyperemia reaching approximately 220% of baseline. In mice, the initial hypoperfusion was pronounced (40-50% of baseline), and the anticipated hyperemic phase barely reached baseline. The duration of hypoperfusion significantly correlated with the duration of the DC shift. As a possible explanation for the pronounced hypoperfusion, mouse cerebral vessels showed enhanced resistance to relaxation by acetylcholine (3 microM) after K+ -induced preconstriction (20, 40, and 80 mM) but dilated normally in response to acetylcholine after preconstriction with U46619, a synthetic thromboxane A2 analog. By contrast, rat vessels dilated readily to acetylcholine after preconstriction by K+. The transient normalization of CBF after hypoperfusion in the mouse was abolished by L-NA but not 7-NI. In summary, the CBF response to cortical spreading depression in mice contrasts with the rat in that the initial hypoperfusion is pronounced, and the hyperemic phase is markedly diminished. The differences in CBF response between species may be in part caused by an increased sensitivity of mouse cerebral vessels to elevated extracellular K+. PMID- 15529019 TI - Spontaneous low-frequency oscillations decline in the aging brain. AB - It is well known that aging leads to a degeneration of the vascular system. Hence, one may hypothesize that spontaneous oscillations decrease in the cerebral microvasculature with aging. Accordingly, the authors investigated the age dependency of spontaneous oscillations in the visual cortex during rest and functional activation. Functional near-infrared spectroscopy was used because it is particularly sensitive to the microvasculature. Visual stimulation led to an increase of oxyhemoglobin, total hemoglobin, and a decrease of deoxyhemoglobin, without any influence of age. Peaks of normalized power spectral density were detected for spontaneous low-frequency (0.07 to 0.11 Hz) and very-low-frequency (0.01 to 0.05 Hz) oscillations, with a higher amplitude for oxyhemoglobin than for deoxyhemoglobin. Spontaneous low-frequency oscillations of oxyhemoglobin and deoxyhemoglobin declined strongly with aging during both rest and visual stimulation. Reduction of spontaneous low-frequency oscillations might indicate a declining spontaneous activity in microvascular smooth muscle cells, in conjunction with an increased vessel stiffness with aging. PMID- 15529021 TI - Role of CETP inhibitors in the treatment of dyslipidemia. AB - PURPOSE OF REVIEW: This review summarizes novel human data on cholesteryl ester transfer protein (CETP) and atherosclerosis and the possible use of CETP inhibitors in the treatment of dyslipidemia. In addition, it will underline that therapeutic targeting of the high-density lipoprotein (HDL) metabolism entails more than simply observing changes in cholesterol levels of this lipoprotein. RECENT FINDINGS: Two pharmacological small-molecule inhibitors of CETP, JTT-705 and torcetrapib, have recently been shown to effectively raise HDL cholesterol in humans without serious side effects when either used as a monotherapy or combined with statins that lower low-density lipoprotein cholesterol. Importantly, prospective data from the Epic-Norfolk study furthermore indicate that elevated CETP concentration in conjunction with elevated triglyceride levels are associated with increased odds for cardiovascular events. Data from the Diabetic Atherosclerosis Intervention Study furthermore show that elevated CETP concentration is associated with increased progression of coronary atherosclerosis in patients with type 2 diabetes who use fenofibrate. SUMMARY: Long-term studies will have to show whether CETP inhibition decreases the risk of atherosclerotic disease in dyslipidemic patients. Increased CETP activity might be detrimental under hypertriglyceridemic conditions which is of importance when considering that a large proportion of patients at increased risk from coronary artery disease exhibit elevated triglyceride levels. Studies into the effects of CETP inhibition in hypertriglyceridemic patients therefore seem warranted. Awaiting the first data on the effect of CETP inhibition on surrogate endpoints for atherosclerosis, this review furthermore outlines that the complexity of HDL metabolism will necessitate a wide variety of studies on many aspects of this intriguing lipoprotein. PMID- 15529022 TI - Intensive statin therapy in acute coronary syndromes: clinical benefits and vascular biology. AB - PURPOSE OF REVIEW: The results of a landmark clinical study comparing intensive statin therapy with conventional statin therapy, in patients with acute coronary syndromes (ACS), are reviewed. The mechanisms behind these results are analysed drawing data from vascular and cell biology. RECENT FINDINGS: The Pravastatin or Atorvastatin Evaluation and Infection Therapy-Thrombolysis in Myocardial Infarction (PROVE IT-TIMI 22) study showed that intensive statin therapy with 80 mg of atorvastatin to achieve a low-density lipoprotein cholesterol of 62 mg/dl resulted in a 3.9% absolute and a 16% relative risk reduction in death or major cardiovascular events up to 2 years, compared to 40 mg of pravastatin, in patients with ACS. The results were especially significant as intensive statin therapy resulted in a very early benefit (<30 days) and occurred against a background of percutaneous coronary intervention (69%) for the index admission and high use of medications for secondary prevention. The PROVE IT and the Myocardial Ischaemia Reduction with Aggressive Cholesterol Lowering (MIRACL) C reactive protein sub-study also showed that atorvastatin (80 mg) resulted in a significant reduction in markers of inflammation, whilst the Reversal of Atherosclerosis with Aggressive Lipid Lowering (REVERSAL) study showed that intensive statin therapy was associated with reduced progression of atherosclerosis compared with conventional doses of statins. SUMMARY: Intensive statin therapy results in a significant early reduction in adverse cardiac events in ACS patients which are sustained over 2 years. The early benefits seen are likely to result from modulation of inflammation, endothelial function and coagulation, i.e. the pleiotropic effects, whereas the greater reduction in low density lipoprotein cholesterol results in reduced long-term events. PMID- 15529024 TI - C-reactive protein as a risk factor versus risk marker. AB - PURPOSE OF REVIEW: C-reactive protein (CRP) is consistently associated with cardiovascular disease in prospective and cross-sectional clinical and epidemiological studies. Inflammation is an important mechanism in cardiovascular disease, and the plasma level of CRP is considered to reflect the inflammatory condition of the patient and/or the vessel wall. In addition, there are also a number of indications for a causal role of CRP in cardiovascular disease. RECENT FINDINGS: A number of new publications show potential causal effects of CRP on cardiovascular disease, and evidence from human-CRP transgenic animals also indicates a causal contribution of CRP to cardiovascular disease. On the other hand, a new large prospective study and an updated meta-analysis indicate that the contribution of CRP to cardiovascular disease is less impressive than reported earlier (odds ratio, 1.58; 95% confidence interval, 1.48-1.68). SUMMARY: We review here the most recent evidence on mechanisms by which CRP is involved as a causal factor in the precipitation of cardiovascular disease. Evidence for such a role is accumulating. PMID- 15529023 TI - Human apolipoprotein A-I and A-I mimetic peptides: potential for atherosclerosis reversal. AB - PURPOSE OF REVIEW: Recent publications related to the potential use of apolipoprotein (apo)A-I and apoA-I mimetic peptides in the treatment of atherosclerosis are reviewed. RECENT FINDINGS: A preliminary report indicating that infusion of apoA-IMilano into humans once weekly for 5 weeks caused a significant decrease in coronary artery atheroma volume has sparked great interest in the potential therapeutic use of apoA-I. Recent studies have revealed that HDL quality (e.g. HDL apolipoprotein and lipid content, including oxidized lipids, particle size and electrophoretic mobility, associated enzymatic activities, inflammatory/anti-inflammatory properties, and ability to promote cholesterol efflux) may be more important than HDL-cholesterol levels. Therefore, when developing new strategies to raise HDL-cholesterol concentrations by interfering with HDL metabolism, one must consider the quality of the resulting HDL. In animal models, raising HDL-cholesterol levels by administering oral phospholipids improved both the quantity and quality of HDL and was associated with lesion regression. An apoA-I mimetic peptide, namely 4F synthesized from D amino acids (D-4F), administered orally to mice did not raise HDL-cholesterol concentrations but promoted the formation of pre-beta HDL containing increased paraoxonase activity, resulting in significant improvements in HDL's anti inflammatory properties and ability to promote cholesterol efflux from macrophages in vitro. Oral D-4F also promoted reverse cholesterol efflux from macrophages in vivo. SUMMARY: The quality of HDL may be more important than HDL cholesterol levels. ApoA-I and apoA-I mimetic peptides appear to have significant therapeutic potential in atherosclerosis. PMID- 15529025 TI - Niacin therapy in atherosclerosis. AB - PURPOSE OF REVIEW: Well designed, randomized, placebo-controlled studies show that niacin prevents cardiovascular disease and death. Unfortunately, early studies and anecdotal evidence have limited its use by promoting the opinion that niacin is intolerable and contraindicated in diabetes. As evidence mounts that treating multiple lipid risk factors decreases cardiovascular risk, the use of niacin in the treatment of atherosclerosis is experiencing somewhat of a renaissance. RECENT FINDINGS: Emerging clinical evidence shows that niacin is both safe and effective in diabetes. Niacin beneficially alters lipoprotein subclass distribution and when used in combination with statins, has additional effects on lipoproteins. Niacin selectively and directly inhibits hepatic diacylglycerol acyltransferase 2, but not diacylglycerol acyltransferase 1, thus inhibiting hepatic triglyceride synthesis and very low density lipoprotein secretion. The recent discovery and characterization of a membrane-bound nicotinic acid receptor (HM74) explains niacin's acute inhibition of adipocyte lipolysis, but the role of HM74 in lowering triglycerides is unclear. Niacin possesses antioxidant, antiinflammatory, and other beneficial effects on atherosclerosis unrelated to lipid lowering. Finally, niacin appears to activate nuclear transcription factors such peroxisome proliferator activator receptor gamma, possibly via prostaglandin metabolism. SUMMARY: New data indicate that niacin alters lipoprotein metabolism in novel ways, and mediates other beneficial nonlipid changes that may be atheroprotective. This information forms the rationale for the use of niacin in combination with agents possessing complementary mechanisms of action (e.g. statins) for cardiovascular risk reduction beyond that observed with monotherapy. Further research into the specific mechanisms of niacin may identify additional targets for future drug development. PMID- 15529026 TI - Cholesterol, statins and dementia. AB - PURPOSE OF REVIEW: Advances in cholesterol biology suggest that cholesterol metabolism modulates beta-amyloid production, and that pharmaceuticals that inhibit cholesterol metabolism might be valuable in therapy of Alzheimer's disease. Although the genetics and cell biology continue to support the link between cholesterol and Alzheimer's disease, recent clinical studies suggest that the animal studies might not directly translate to clinical studies in humans. RECENT FINDINGS: This review will highlight advances in genetics, cell biology and clinical sciences investigating the relationship between cholesterol and Alzheimer's disease. SUMMARY: Cholesterol, its catabolites and proteins that regulate cholesterol levels all modulate processing of amyloid precursor protein. Statins hold promise in therapy of Alzheimer's disease, but the current data are more consistent with a model of statins that act as neuroprotective agents rather than inhibitors of beta-amyloid production. PMID- 15529028 TI - Bibliography. Current world literature. Therapy and clinical trials. PMID- 15529027 TI - An update on alcohol and atherosclerosis. AB - PURPOSE OF REVIEW: Epidemiological studies consistently link moderate alcohol use with a lower risk of cardiovascular disease, but a number of important issues remain controversial. These include the putative impact of non-alcoholic constituents of some alcoholic beverages, the role of genetic factors, potential mechanisms for this association, and confirmation of the relationship in experimental models. RECENT FINDINGS: Although high-density lipoprotein cholesterol (HDL-C) is considered the primary mediator of the cardiovascular effects of moderate drinking, recent evidence has shown the alcohol-HDL-C relation is not linear beyond the range of moderate drinking. Moderate alcohol use also has important inverse relations with inflammatory factors. Some, but not all, animal models confirm the anti-atherogenic effects of ethanol and highlight inflammatory factors as one possible mechanism. The non-alcoholic constituents of red wine also have anti-atherogenic and perhaps even life-extending properties in vitro, but their relevance to humans remains uncertain. Genetic variants of the apolipoprotein E and interleukin 6 genes in humans may modify how alcohol influences atherosclerosis, further emphasizing the importance of HDL-C and inflammatory factors as mediators. SUMMARY: The robust relationship between moderate drinking and lower risk of cardiovascular disease remains an intriguing area of investigation. Clarifying potential gene-environment interactions and translational research into uses for non-alcoholic components will be important areas for future investigation. PMID- 15529029 TI - Nutrition and metabolism. PMID- 15529030 TI - Genetics and molecular biology: genetic epidemiology. PMID- 15529031 TI - Lipid metabolism: is liver X receptor (LXR) a regulator of adipocyte differentiation? Consequences of stearoyl-CoA desaturase activation by LXR. PMID- 15529032 TI - Hyperlipidaemia and cardiovascular disease. PMID- 15529033 TI - Therapy and clinical trials: clinical therapy and trials. PMID- 15529056 TI - New high blood pressure guidelines create new at-risk classification: changes in blood pressure classification by JNC 7. AB - High blood pressure has become increasingly prevalent and is an important risk factor for cardiovascular disease. The Seventh Report of the Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) has redefined normal blood pressure as less than 120/80 mm Hg and created a new blood pressure category called "prehypertension" for those with a systolic BP of 120 to 139 mm Hg or a diastolic BP of 80 to 89 mm Hg. This new blood pressure category was created to identify adults considered to be at risk for developing hypertension and to alert both patients and healthcare providers of the importance of adopting lifestyle changes. Recognition of prehypertension provides important opportunities to prevent hypertension and cardiovascular disease. PMID- 15529058 TI - New blood thinner offers first potential alternative in 50 years: ximelagatran. AB - Traditional anticoagulants employed in the treatment of thrombosis include the injectable heparins and oral warfarin. Though effective, these traditional agents are fraught with limitations in their ease of use in the clinical setting. Warfarin, for example, has many pharmacokinetic properties and food-and-drug interactions that result in unpredictable patient response and the need for expensive and time-consuming monitoring of coagulation status. Ximelagatran is a novel, promising, orally active, direct thrombin inhibitor currently in development that, for the first time in 50 years, offers a potential alternative to the mainstay oral agent "warfarin." Advantages of ximelagatran over warfarin include predictable pharmacokinetics and pharmacodynamics, a broad therapeutic window, no routine anticoagulant monitoring, no clinically significant drug interactions, and fixed-dose administration. Ximelagatran has been evaluated for thromboprophylaxis following orthopedic surgery, acute treatment and secondary prevention of venous thrombosis, stroke prevention in atrial fibrillation, and acute coronary syndromes. Results of clinical trials suggest that ximelagatran is equally or more efficacious than warfarin and/or low-molecular-weight heparin therapy without increasing rates of minor or major bleeding. Although postmarketing surveillance will provide the final test of this drug, the future looks promising for addition of a new anticoagulant with the potential to provide excellent efficacy, predictable response, and reduced adverse effects. Pending regulatory approval, ximelagatran may help overcome barriers to appropriate anticoagulant therapy, thereby decreasing morbidity and mortality associated with thrombotic diseases. PMID- 15529059 TI - Public defibrillation: increased survival from a structured response system. AB - The Public Access Defibrillation (PAD) trial was a prospective, randomized, controlled study designed to compare the number of persons surviving to hospital discharge after experiencing an out-of-hospital cardiac arrest (OOH-CA) among "community units" randomized to receive cardiopulmonary resuscitation (CPR) only or CPR plus an automated external defibrillator (AED). In 24 centers across the United States and Canada, 993 community units, composed of 1260 individual facilities, trained more than 19,000 layperson responders in CPR-only or CPR+AED. Survival to hospital discharge in the CPR+AED arm was double that of the CPR-only arm (30 vs 15, P = .03; RR = 2.0, 95% CI [1.07-3.77]). Intense focus on facility infrastructure, including responder recruitment and training, communication, evaluation, and oversight, was necessary for implementing the emergency response systems for the trial. Use of an AED within this structured response system can increase the number of survivors to hospital discharge after OOH-CA. Trained nonmedical responders can use AEDs safely and effectively. PMID- 15529060 TI - New treatment option for heart failure patients: eplerenone. AB - Aldosterone plays an important role in the harmful cardiac remodeling process and pathophysiology of heart failure after a myocardial infarction. Until recently, spironolactone (Aldactone) was the only pharmacologic agent available to directly block the deleterious effects of aldosterone. The use of spironolactone is complicated by its antiprogesterone and antiandrogen side effects, such as gynecomastia and menstrual irregularities. Eplerenone (Inspra), a member of a new class of drugs called selective aldosterone receptor antagonists, was recently approved for the treatment of both hypertension and post-myocardial infarction heart failure and appears to be devoid of the antiprogesterone and antiandrogen effects. In a trial in patients with heart failure following a myocardial infarction, eplerenone treatment significantly reduced mortality and morbidity compared to placebo. Eplerenone may be considered as part of the therapeutic plan in patients who have suffered a myocardial infarction and demonstrate evidence of heart failure. PMID- 15529061 TI - Heal thyself: Potential applicability of stem cell therapy in the management of heart disease. AB - Acute myocardial infarction results in regional necrotic heart tissue that is considered irreversible. Although angioplasty and thrombolytic therapy can remove the offending atherosclerotic plaque and thrombi, both therapies are dependent upon timely recognition and initiation of treatment and thus have a limited window of opportunity. No currently available therapy has the capability to restore cardiomyocytes or to replace myocardial scar tissue with contractile tissue. In animal models, use of a wide range of cells such as fetal cardiomyocytes, skeletal myoblasts, and bone marrow stem cells have been shown to differentiate into functional cardiomyocytes. In addition, transplantation of adult stem cells directly into the area of infarction has shown clinical promise. This article explores the current data on extramedullary hematopoiesis, stem cell differentiation, and stem cell therapy and its ability to repair injured or ischemic cardiac tissue. PMID- 15529062 TI - Drug-coated stents: preventing restenosis in coronary artery disease. AB - Since its introduction in 1977, the success of percutaneous interventional cardiology has been limited by the occurrence of restenosis. Drug-eluting stents, particularly sirolimus- and paclitaxel-coated stents, have been shown in randomized controlled trials to dramatically reduce restenosis in single, de novo, native coronary arteries. Over the last 2 years, investigators have reported that these stents can also reduce restenosis in more complex patient situations such as in diabetics, during acute coronary syndromes, in long atherosclerotic lesions and small arteries, and even after in-stent restenosis. These outcomes increase the clinical value of this technology to "real world" practice. This article reviews the current state of our knowledge regarding drug eluting stents and identifies areas for further research. PMID- 15529063 TI - Newly mapped gene for thoracic aortic aneurysm and dissection. AB - Thoracic aortic aneurysm and dissection (TAAD) is associated with high mortality and medical expense. These poor outcomes are preventable by surgical repair; however, identifying at-risk individuals is difficult. Researchers are actively surveying the human genome (the repository of human genes) to characterize the genetic determinants of TAAD by identifying chromosomal regions likely to harbor such predisposing genes. In previous studies, investigators identified genetic markers shared by a subset of families who were ascertained to have the disease, which clustered into 2 chromosomal regions: 5q13-q15 (TAAD1) and 11q23.2-q24 (familial aortic aneurysm [FAA1]). In a subsequent study, a third chromosomal region at 3p24-25 (TAAD2) was found to contribute to TAAD in a 4-generation, 52 member family that displayed little evidence of sharing either the TAAD1 or FAA1 regions. Although additional regions of the genome may contribute to TAAD, investigators are focusing their efforts on identifying the actual genes and the specific mutations that participate in the disease process. The goal of these endeavors is to develop screening tests to identify individuals at risk for familial TAAD. This genetic discovery has significant clinical implications because high-risk individuals and families can be closely monitored and can benefit from preventative surgical repairs. PMID- 15529065 TI - A shot of good cholesterol: synthetic HDL, a new intervention for atherosclerosis. AB - One of the American Heart Association's Top 10 Research Advances for the Treatment of Heart Disease is the use of a synthetic form of high-density lipoprotein (HDL) to reduce coronary atherosclerosis (JAMA. 2003;290:2292-2300). While HDL has not been a target for therapy for dyslipidemias, new insight into the major protein component of HDL, apolipoprotein A-I, may lead to new therapies. Apolipoprotein A-I was recently found to be a better predictor of cardiovascular events than is low-density lipoprotein (Am Heart J. 2003;146:227 233; J Intern Med. 2004;255:188-205). This article reviews the recent study by Nissen and colleagues describing the finding of a genetic mutation in HDL in some persons in Italy and the subsequent development of a synthetic form of HDL to be used as an infusion to successfully target atherosclerotic lesions (JAMA. 2003;290:2292-2300). In addition, controversies related to HDL cholesterol as a target for therapy are reviewed. Implications for nursing research, education, and practice are also described. PMID- 15529064 TI - Vampire bats yield potent clot buster for ischemic stroke. AB - While tissue-type plasminogen activator (tPA) is currently the standard "clot busting" drug used to treat patients with acute ischemic stroke, another new option for treatment is now under clinical study. Desmodus rotundus plasminogen activator (DSPA), an enzyme found in the saliva of vampire bats, may offer another therapeutic option instead of tPA, a current therapy for stroke. Animal research indicates that tPA may mediate neuronal death as well as increase systemic plasminogen consumption and fibrinogenolysis. Conversely, DSPA's activity is dependent on the presence of fibrin and therefore has not been associated with the systemic plasminogen consumption and fibrinogenolysis that potentially may occur in those receiving tPA. In animal studies, tPA was found to exhibit "inherent neurotoxic properties" not seen with DSPA. In addition, DSPA may be administered up to 9 hours after the onset of symptoms, unlike tPA, which cannot be given after 3 hours without potential risk of additional brain injury. Phase II clinical trials have demonstrated a positive result in human subjects. Phase III trials are currently under way in stroke populations. PMID- 15529066 TI - New blood test to measure heart attack risk: C-reactive protein. AB - In 2003, the American Heart Association and the Centers for Disease Control and Prevention issued a scientific statement that suggested the use of high sensitivity C-reactive protein, a recognized marker of inflammation, as an optional myocardial infarction risk factor measurement (Circulation. 2003;107:499 511). Compared with earlier guidelines, the statement suggested a more aggressive approach for identifying patients at risk of cardiovascular events. The increased interest in and the mounting evidence of the role of inflammation in the development of acute coronary syndrome was a major impetus for this. This article focuses on the use of high-sensitivity C-reactive protein as a nonlipid cardiovascular event risk marker and the implications for practice, education, and research. PMID- 15529067 TI - Over-the-counter statins? Sorting out the issues. PMID- 15529069 TI - Ventricular dysrhythmias in heart failure. AB - Left-ventricular systolic dysfunction, or heart failure (HF), is a chronic, progressive condition with a poor prognosis. Approximately 50% of deaths, especially in mild to moderate cases, are sudden. Most sudden deaths are thought to be due to ventricular tachycardia; however, premature ventricular contractions and couplets parallel severity of HF and have been associated with increased mortality risk as opposed to dysrhythmic death. Ventricular arrhythmogenesis results from many mechanisms (afterdepolarizations, reentry, and enhanced automaticity) and preconditions (electrophysiologic abnormalities, neuroendocrine activation, electrolyte imbalances, scar from an ischemic event in ischemic cardiomyopathy, fibrosis in dilated cardiomyopathy, hemodynamic abnormalities, and HF medical management). Nurses are key caregivers in optimally managing HF, either by direct actions or by using advocacy, communication, and collaboration skills to promote positive outcomes. Ventricular dysrhythmia management consists of facilitating core HF pharmacologic and nonpharmacologic medical therapies, using amiodarone to improve symptoms, as needed, and utilizing implantable cardioverter-defibrillator therapy to reduce the risk of sudden cardiac death. PMID- 15529070 TI - Managing the heart failure patient with obstructive lung disease. AB - Heart failure (HF) and obstructive lung disease (OLD) are chronic diseases associated with frequent emergency department visits and repeated hospitalizations and, when linked comorbidly, strongly associated with poor prognostic outcomes. When dyspnea occurs in a HF patient with OLD, the practitioner is challenged to determine which disease exacerbation underlies the current symptom. The purpose of this article is to provide the HF practitioner with practical information about how to make these distinctions and how to develop a treatment plan for the HF patient that incorporates important information about OLD. The pathophysiology of OLD and HF is reviewed first, followed by details on the assessment of dyspnea and the management of acute exacerbations of OLD. Finally, pharmacologic issues in the management of concomitant HF and OLD are reviewed. PMID- 15529071 TI - Atrial fibrillation in the heart failure population. AB - Atrial fibrillation (AF) is the most common chronic arrhythmia. Incidence of AF in the heart failure population is high--10% to 30%--and is associated with a reduction in exercise capacity and a poor long-term prognosis. Management of AF includes 4 strategies: control of heart rate, termination of AF/conversion back to sinus rhythm, prevention of recurrences, and prevention of thromboembolic events. This article reviews these management strategies in the heart failure population. PMID- 15529072 TI - Type 2 diabetes and chronic systolic heart failure: clinical implications. AB - Diabetes is a chronic progressive endocrinopathy associated with significant macrovascular and microvascular complications as well as cardiomyopathy and heart failure (HF). Diabetes and chronic systolic HF result in similar activation of pathologic neurohormonal pathways. When diabetes and HF coexist, morbidity and mortality significantly increase. This article reviews important clinical issues in the care of patients with diabetes and HF. A review of pertinent pathophysiologic principles is provided, followed by a discussion of the treatment issues related to this population. Treatment issues include vascular disease risk factor modification, HF pharmacotherapy, glycemic management, and control of other common comorbid conditions. PMID- 15529074 TI - Identifying and treating depression in patients with heart failure. AB - Depression is a common comorbid condition in patients with heart failure (HF) that often goes undiagnosed and untreated. Unless symptoms of depression are specifically looked for, they can be easily missed or mistaken for HF. Clinical depression is a syndromal diagnosis based on patient history, the report of signs and symptoms, and the exclusion of competing diagnoses. This article describes the recommended strategies for recognizing and diagnosing depression. General, psychotherapeutic, and pharmacological approaches to treatment of depression in HF are also discussed. General treatment measures include optimization of medical therapy, patient education, exercise, social support, and family care. Although there are many types of psychotherapy, cognitive-behavioral therapy is particularly appropriate for patients with HF and is described in this article. Categories of antidepressant drugs, including serotonin reuptake inhibitors, tricyclic antidepressants, and miscellaneous antidepressants, are reviewed relative to treating depression in patients with HF. Given the high prevalence and adverse impact of untreated depression in patients with HF, it is essential for advanced practice nurses to develop the requisite knowledge and skills for the identification and treatment of depression. PMID- 15529075 TI - Anemia in heart failure: implications, management, and outcomes. AB - Despite advances in the treatment of heart failure (HF) over the past decade, the prognosis remains poor. Anemia is a well-recognized comorbidity in many chronic conditions, but its role in HF has only recently been recognized. Anemia is significantly related to symptoms, exercise capacity, and prognosis in HF; it has been identified as an independent risk factor for mortality in those with left ventricular dysfunction. When HF patients have concomitant renal disease, they invariably become anemic owing to erythropoietin deficiency. In chronic HF patients without renal disease, erythropoietin levels may be elevated in response to anemia, but not adequately increased to overcome it. Some degree of erythropoietin resistance may also be present because of elevated plasma levels of cytokines. Several studies in anemic HF patients have shown positive outcomes using erythropoietin and iron supplementation therapy to increase hemoglobin concentrations to more normal levels. This article reviews the current information available regarding anemia in HF and discusses the clinical implications and treatment of this syndrome. PMID- 15529076 TI - The heart of sleep: sleep-disordered breathing and heart failure. AB - Heart failure is associated with high rates of hospitalization and mortality as well as great economic burden in the United States. Recent data show that a high percentage of patients with depressed left ventricular ejection fractions suffer from sleep-disordered breathing, contributing to this incidence of morbidity and mortality. Since the signs and symptoms associated with sleep disorders do not differ significantly from chronic heart failure signs and symptoms, detection and recognition of sleep-disordered breathing is a clinical challenge. Thus clinicians frequently fail to recognize it as a possible contributor to the development of heart failure or as a consequence of the disease. This article discusses sleep, sleep-disordered breathing, its effects and consequences on the cardiovascular system, strategies for identifying at-risk individuals, and treatment options. PMID- 15529077 TI - Managing noncardiac pain in heart failure patients. AB - Both acute and chronic pain are common coexisting problems in patients with heart failure. Because nonsteroidal anti-inflammatory drugs and corticosteroids are contraindicated in heart failure, traditional pain management algorithms require modification. This article reviews pertinent pain management principles, including pain vocabulary, barriers to pain management, and general pain assessment and treatment measures. Issues unique to the heart failure patient are discussed and specific interventions for the heart failure patient with acute or chronic pain are then delineated. PMID- 15529078 TI - [What's the best curriculum for a future vascular physician?]. PMID- 15529079 TI - [Specific objectives of initial training in vascular medicine]. PMID- 15529080 TI - [Accreditation criteria for hospital wards necessary to validate initial training in vascular medicine]. PMID- 15529081 TI - [Can dental avulsions be performed under anticoagulation?]. PMID- 15529082 TI - [Antiretroviral therapy and cardiovascular risk]. AB - Current antiretroviral therapy protocols enable long-term survival of HIV infected patients, decreasing the risk of infectious complications. Three classes of anti-HIV treatments are available. With longer survival, unusual cardiovascular complications related to iatrogenic biological anomalies (dyslipidemia and impaired glucose tolerance) have appeared among this young population which is exposed to usual risk factors of atherosclerosis. Antiretroviral therapies are suspected to cause these complications, inducing maturity-onset diabetes in 4 to 20% of patients, impaired glucose tolerance in 15 to 60%, hypertriglyceridemia in 15 to 74% depending on the survey, and hypercholesterolemia in 20 to 60%, especially in case of associated lipodystrophia. A lipid battery including total cholesterol, HDL, and triglycerides, and 12-h fasting blood glucose should be obtained before initiating antiretroviral therapy. Any anomalous finding should be followed carefully with regular surveillance every 3 to 6 months and search for other causes of secondary dyslipidemia. In the event of casual and persisting elevation of LDL-cholesterol levels, a statin treatment can be introduced. For secondary prevention, irrespective of the context, recommendations currently merge with the consensus applying to the general population. These patients require careful surveillance of cardiovascular risk factors and a specific care in addition to treatment of their immunodeficiency. PMID- 15529083 TI - [Puffy hand in long-term intravenous drug users]. AB - Narcotic addiction may induce systemic and local complications. Intravenous injections of drugs can cause venous thrombosis, and septic or embolic complications. The puffy hand sign is a more uncommon complication of hard-core injection addicts. Three long-term intravenous drug users, two males, one female, mean age 30.6 years (26-37) presented puffy hands. These patients had been drug addicts for four to twelve years (mean duration 7.3 years) and had stopped heroin injections for 3-5 years (mean 4.6), participating in a buprenorphine substitution program. The edema appeared several years after drug cessation (1.5 5, mean 2.3). Typically the puffiness was bilateral, the hands swollen from the proximal segments of the fingers to the wrist. In one patient, the edema was localized both in the hands and in the feet. The edema was not pitting and unaffected by elevation. Duplex ultrasound examination of the extremities was normal. Lymphangiography performed in one patient was consistent with deep lymphatic destruction. Puffy hand syndrome appears to be the end result of lymphatic obstruction. Repeated injections of drugs in or outside the veins destroy the lymphatics. Buprenorphine may play an important role in the puffy hand sign. Although it is supposed to be administered orally, many drug addicts use it as an i.v. solution. Because buprenorphine is poorly soluble, it causes lymphatic obstruction. This type of hand for which no therapy exists must be differentiated from deep palmar space infection with dorsal edema which requires incision and drainage. PMID- 15529084 TI - [Imaging of an aberrant occipital artery arising from the extracranial segment of the internal carotid artery]. AB - We report a case of asymptomatic occlusion of the right internal carotid artery owing to the presence of a rare anatomic variant (occipital artery originating from the extracranial segment of the internal carotid artery). During the preoperative workup for a coronary artery bypass grafting (CABG) in a 65-year-old patient, an occlusion of the right internal carotid artery was visualized on Duplex ultrasound. Distally to the occluded site, the artery was patent thanks to a vessel parallel to it but with a reverse blood flow. MRA and angiography of the brachiocephalic trunk confirmed the occlusion of the internal carotid artery and its opacification distally via an aberrant occipital artery arising at the supra bulbar level and communicating with the right vertebral artery. Therapeutic abstention was decided and the patient was operated of CABG without any neurological complication. Occipital artery arising from the internal carotid artery is a rare anomaly. Its association with internal carotid artery occlusion may be beneficial because it makes surgery possible, even in a chronic stage, if the patient is symptomatic. If the patient is asymptomatic, therapeutic abstention could be considered. PMID- 15529085 TI - [Angioplasty with stenting for buttock claudication]. AB - We report the case of a patient presenting severe buttock claudication with normal neurologic and osteoarticular exams. He underwent a guidewire recanalization of his occluded superior gluteal artery followed by a percutaneous angioplasty with stenting, resulting in total relief of symptoms. This observation represents the first publication describing the use of a stent with recanalization of the gluteal artery. The technique seems promising for buttock pathology. PMID- 15529086 TI - [Aneurysm of the superior gluteal artery]. AB - We report the case of an aneurysm of the superior gluteal artery that occurred more than ten years after a pelvic trauma. The diagnosis of this uncommon condition is sometimes difficult because of the predominant neurologic symptoms. Rupture can be life-threatening. The patient was treated by an embolization; thereafter a surgical procedure with control of the internal iliac artery and an endoaneurysmorrhaphy was performed. The main characteristics of this pathology are indicated. PMID- 15529087 TI - Bipolar disorder: a national health concern. AB - Bipolar disorders are prevalent, disabling, and costly diseases that often pursue an inexorable course. Underdetection, misdiagnosis, and diagnostic delay frequently and unnecessarily interfere with appropriate treatment of the disorder. Mortality studies in bipolar disorder underscore the relevance of both unnatural and natural causes of death, inviting the need for improved preventative and primary health care for bipolar patients. The treatment framework for bipolar disorder must recognize and anticipate the multidimensionality and comorbidity of this illness. Pharmacotherapy is necessary, with multiple concomitant medications required for most patients. In addition, adjunctive psychosocial interventions offer enhanced compliance and may beneficially influence psychopathological and functional outcomes. This article emphasizes the public health concern of bipolar disorder, and provides tactics to enhance detection of cryptic bipolar states, underscore the clinical and pathophysiological relevance of comorbidity in bipolar disorder, and provide a framework for multimodality therapy for this condition. PMID- 15529089 TI - Safety considerations in pharmacotherapy of bipolar disorder. AB - Bipolar disorder is a chronic, frequently relapsing illness with a prevalence of 1.2% to 3.4% in the general population. It is associated with high disability, higher comorbidity due to medical illnesses, and significant social and economical consequences for patients, their families, and society. The episodic nature of this disease warrants rational use of medications and proper monitoring for adverse events. Various drug classes, such as mood stabilizers, antipsychotics, benzodiazepines, and antidepressants, are used for the acute and maintenance treatment of bipolar disorder. Each group of drugs is associated with wide array of adverse events and drug interactions, which are the main hurdles in treatment outcome and compliance. Common side effects seen with several agents, particularly antipsychotics, are somnolence, weight gain, extrapyramidal symptoms, dyslipidemia, type-2 diabetes, and hyperprolactinemia. Major drug interactions are seen with drugs such as carbamazepine, due to hepatic enzyme induction. Adverse effects such as somnolence are tolerability concerns and can be managed easily; others, such as diabetes mellitus, are safety concerns. It is prudent to have precise knowledge of the individual drug's side-effect profile, pharmacokinetics, and pharmacodynamics, to plan a treatment regimen. More research is needed to understand potential risks of various drugs and to devise and incorporate monitoring protocols in the treatment regimen. PMID- 15529090 TI - The role and impact of psychotherapy in the management of bipolar disorder. AB - A growing body of evidence documents the value of structured psychotherapeutic interventions for the co-management of bipolar disorder in the context of ongoing medication treatment. This article reviews the rationale, elements, and outcomes for those psychosocial treatments for bipolar disorder that have been examined in randomized trials. The available evidence suggests that interventions delivered in individual, group, or family settings, can provide significant benefit to patients undergoing pharmacotherapy for bipolar disorder. PMID- 15529091 TI - Drugs for migraine. PMID- 15529092 TI - Drugs for diabetes. PMID- 15529093 TI - Drugs for tobacco dependence. PMID- 15529094 TI - Drugs for psychiatric disorders. PMID- 15529095 TI - Drugs for lipid disorders. PMID- 15529096 TI - Drugs for pneumonia. PMID- 15529097 TI - Drugs for assisted reproduction. PMID- 15529098 TI - Drugs for allergic disorders. PMID- 15529099 TI - Diet, drugs and surgery for weight loss. PMID- 15529100 TI - Drugs for asthma. PMID- 15529101 TI - Drugs for prevention and treatment of postmenopausal osteoporosis. PMID- 15529102 TI - Cardiovascular drugs in the ICU. PMID- 15529103 TI - Drugs for rheumatoid arthritis. PMID- 15529105 TI - Drugs of choice for cancer. PMID- 15529104 TI - Drugs for hypertension. PMID- 15529106 TI - Drugs for treatment of heart failure. PMID- 15529107 TI - Drugs for epilepsy. PMID- 15529108 TI - Drugs for HIV Infection. PMID- 15529109 TI - Drugs for peptic ulcers. PMID- 15529110 TI - Choice of antibacterial drugs. PMID- 15529111 TI - Antimicrobial prophylaxis for surgery. PMID- 15529112 TI - Advice for travelers. PMID- 15529113 TI - Drugs for Parkinson's Disease. PMID- 15529114 TI - Drugs for pain. PMID- 15529115 TI - Choice of contraceptives. PMID- 15529116 TI - Drugs for sexually transmitted infections. PMID- 15529117 TI - Drugs for cardiac arrhythmias. PMID- 15529118 TI - Synapsin and synaptic vesicle protein expression during embryonic and post-natal lens fiber cell differentiation. AB - PURPOSE: Reorganization of cytoskeleton and membrane biogenesis are dynamically coordinated during lens fiber cell differentiation and development to produce an organ with precise dimensions and optical properties. Cargo vesicle trafficking is fundamental to cell elongation and has also been implicated in degenerative disease mechanisms. Alzheimer precursor protein (AbetaPP) acts with kinesin, synapsin, and synaptic vesicle proteins to mediate cargo vesicle transport and membrane fusion in neurons. In our previous studies we demonstrated that AbetaPP is also a key element in lens fiber cell formation, and in early-onset cataract that occurs along with early-onset Alzheimer disease in Down syndrome. In the present study we examine lens expression and regulation of a complement of genes associated with cargo and synaptic vesicle transport in neurons. METHODS: RT-PCR, immunoblot, and immunohistochemical methods were used to characterize expression of AbetaPP and kinesin associated motor proteins, synapsins, and synaptic vesicle proteins in mouse and rat embryonic, post-natal, and adult lenses. Phospho specific anti-synapsin antibodies were used to determine the distributions of site-1 phosphorylated and dephosphorylated synapsin protein. RESULTS: We demonstrate that a substantial complement of cargo and synaptic vesicle proteins involved in AbetaPP mediated vesicle transport are expressed in lenses along the anterior-posterior axis of fiber cells in embryonic and adult lenses, consistent with vesicles, actin filaments, and neuron-like arrangement of microtubules in lenses shown by others. We identify temporal regulation of synapsins I, II, and III during embryonic and post-natal lens development consistent with their roles in neurons. Regulation of vesicle cytoskeleton attachment, actin polymerization, and the capacity to stimulate cell differentiation by synapsins are governed in large part by phosphorylation at a conserved Ser9 residue (site-1). We demonstrate discrete distributions of Ser9 phospho- and dephospho-synapsins along the axial length of rapidly elongating embryonic lens fiber cells, and decreased levels of site-1 phosphorylated synapsins in adult lenses. CONCLUSIONS: The present findings demonstrate several fundamental parallels between lens and neuron vesicle trafficking cell biology and development, and suggest that more extensive AbetaPP related vesicle trafficking disease mechanisms may be shared by lens and brain. PMID- 15529119 TI - Give me a place to stand and I will move the earth. PMID- 15529120 TI - Radiolucency in the mandible. PMID- 15529121 TI - Liver disease: current perspectives on medical and dental management. PMID- 15529122 TI - Gingival melanin pigmentation and its treatment with the CO2 laser. AB - OBJECTIVE: There are only a few reports in the literature evaluating the effects of the carbon dioxide (CO2) laser on gingival melanin pigmentation (GMP). This study was performed to evaluate the effects of the CO2 laser at superpulse mode, applied for gingival depigmentation. STUDY DESIGN: GMP at the anterior maxillary and mandibular gingiva of 10 patients was treated using the superpulsed CO2 laser (10 watts, 0.8 mm spot size, 20 Hz, 10 milliseconds). Pigmented areas were measured on pre- and postoperative standard digital images by the aid of an image analyzing software. Statistical analysis of the data was performed by Mann Whitney U test. RESULTS: Ablation of the hyperpigmented gingiva was accomplished with minimal carbonization and almost no bleeding. Postoperative healing was uneventful with no significant postoperative pain. Two cases of partial repigmentation were observed during 24-month follow-up. Statistical analysis of the data revealed a significant difference between pre- and postoperative measurements of pigmented area. CONCLUSION: Application of the superpulse mode of CO2 laser appears to be an effective and safe method for the elimination of GMP. PMID- 15529123 TI - Blood-flow change and recovery of sensibility in the maxillary dental pulp during and after maxillary distraction: a pilot study. AB - OBJECTIVE: To examine the change in blood flow and recovery of sensibility in the maxillary dental pulp during and after maxillary distraction. STUDY DESIGN: The subjects included 5 patients undergoing high Le Fort I osteotomy and maxillary distraction (D-group) and a reference group of 14 patients who underwent a common single-segment Le Fort I osteotomy, 1-stage maxillary advancement, and mandibular setback surgery (C-group). Eleven (D-group) and 54 (C-group) maxillary incisors were assessed preoperatively and at 1-7 days, 14 days, and 3 months postoperatively. Pulpal blood flow (PBF) was measured by laser Doppler flowmetry, and pulpal sensibility (PS) was investigated by electrodiagnostics. RESULTS: From postoperative days 1 to 5 (the latency period), the PBF tended to be higher in the D-group than in the C-group. From day 6 to 3 months postoperatively (during and after maxillary distraction), the PBF values of the 2 groups were similar. The PS remained negative up to 14 days postoperatively in both groups. However, at 3 months after the operation, a higher proportion (90.9%) of teeth in the D group was positive for PS. CONCLUSION: These results suggest that maxillary distraction is a favorable technique for maintenance of PBF and recovery of PS in the maxillary teeth after surgery. PMID- 15529124 TI - Relationship of calcitonin gene-related peptide in synovial tissues and temporomandibular joint pain in humans. AB - OBJECTIVE: To elucidate the expression of calcitonin gene-related peptide (CGRP) in synovial tissue taken from the human temporomandibular joint (TMJ) with internal derangement, and discuss the relationship between CGRP and joint pain. STUDY DESIGN: Using an immunohistochemical technique, 48 joints in 48 patients were examined. As controls, synovial tissue specimens from 7 joints with habitual dislocation without pain were also examined. RESULTS: In all of the internal derangement and control subjects, CGRP-positive cells were observed in the connective tissues around the blood vessels beneath the lining cells. The extent score of CGRP was significantly higher in the internal derangement group than in the control group (P=.033). There was a significant positive correlation between the extent score of CGRP and joint pain (P=.036, r=0.30). CONCLUSIONS: These results suggest that the expression of CGRP is increased in the synovial tissues from patients with internal derangement, and that CGRP seems to play an important role in the mechanism of pain production in patients with symptomatic internal derangement. PMID- 15529125 TI - Maxillary distraction resulting in facial advancement at Le Fort III level in cleft lip and palate patients: a report of two cases. AB - An 18-year-old female and a 14-year-old male who had previously received surgery for primary repair of a nonsyndromic cleft lip and palate (including alveolar defect bone grafting) unintentionally developed facial advancement at the Le Fort III level after surgical correction of their maxillary hypoplasia. The Le Fort I osteotomy, originally performed for their maxillary dentoalveolar hypoplasia, was an incomplete osteotomy. It was performed without down-fracture, leaving the pterygomaxillary and septal junctions intact. The gradual advancement of the maxilla during distraction osteogenesis was planned to correct the hypoplastic maxilla, and also prevent subsequent hypernasality; however, during the distraction procedure by means of a rigid external device both patients developed an unintentional facial advancement at the Le Fort III level. PMID- 15529126 TI - Carcinoma of the tongue: a case series analysis of clinical presentation, risk factors, staging, and outcome. AB - OBJECTIVE: Oral cancer is primarily a disease of epithelial origin. The tongue is the most common site of oral cancer. Oropharyngeal cancer accounts for approximately 2% of all cancer deaths. The purpose of this study was to analyze a case series of patients with tongue cancer. STUDY DESIGN: The medical records of 322 consecutive patients diagnosed with squamous cell carcinoma (SCC) of the tongue treated from 1979-1994 were reviewed. RESULTS: Patients had a mean age of 61.1 years and a 1.5:1 male to female ratio. Approximately, half of the cancers were diagnosed at an advanced stage and most involved the base of the tongue. The majority of oral tongue tumors were diagnosed at an early stage and had more well differentiated cell types than those of the base of the tongue. Localized discomfort was the most common complaint (66.5%), present for up to 6 months in patients prior to diagnosis and were related to the oral tongue in 90% of cases. Symptoms associated with base of tongue lesions included neck masses, dysphagia, ear pain, and weight loss. Seventy-nine percent of the patients smoked, 58% consumed alcohol on a daily basis, and 43% consumed more than 4 alcoholic drinks daily. Primary radiotherapy was the treatment of 49.6% lesions, of which 57% were at an early stage at diagnosis. The mean overall survival was 3 years and 5 months, with a 5-year overall survival of 40%. Patients with stage 1 tumors had a disease-specific survival of over 80%. The 5-year survival of patients with cancer of the oral tongue was 43% and cancer of the base of the tongue was 27%. CONCLUSION: The prognosis is poorer for patients presenting with advanced stage and with tumors involving the base of the tongue. Symptoms were more common in tumors of the oral tongue, which likely results in earlier diagnosis. Patients with SCC of the base of the tongue presented with advanced stage of disease and were typically poorly differentiated lesions. PMID- 15529127 TI - Healing of oral lichenoid lesions after replacing amalgam restorations: a systematic review. AB - OBJECTIVE: We sought to systematically review the literature related to oral lichenoid lesions (OLLs) and amalgam restorations. STUDY DESIGN: Cohort and case controlled studies (no randomized controlled trials or controlled clinical trials available) were reviewed with respect to inclusion criteria and data on patients with OLLs, treatment interventions, and the measurement of outcomes. RESULTS: Fourteen cohort and 5 case-controlled trials met the criteria. The study population consisted of 1158 patients (27% male and 73% female; age range, 23-79 years). From 16% to 91% of patients had positive patch test results for at least 1 mercury compound. Of 1158 patients, 636 had to have their restorations replaced. The follow-up period ranged from 2 months to 9 1/2 years. Complete healing ranged from 37.5% to 100%. The greatest improvements were seen in lesions in close contact with amalgam. CONCLUSIONS: Protocols must be standardized to obtain valid results. The replacement of amalgam restorations can result in the resolution or improvement of OLLs. Patch testing seems to be of limited value. The topographic relationship between an OLL and an amalgam restoration is a useful--but not conclusive--marker. PMID- 15529128 TI - Dyskeratosis congenita vs. chronic graft versus host disease: report of a case and a review of the literature. AB - Dyskeratosis congenita (DC) is a rare inherited bone marrow failure syndrome characterized by the triad of reticular pigmentation of the skin, nail dystrophy, and mucosal leukoplakia. DC is often associated with severe pancytopenia, and bone marrow failure is the principal cause of early mortality. Malignant transformation of mucosal leukoplakias may also occur. Rarely, aplastic anemia precedes the other clinical manifestations of the disease. We present a case of a 13-year-old boy who was diagnosed at age four with idopathic aplastic anemia, was treated successfully with an allogeneic bone marrow transplant, then subsequently developed skin, nail, and tongue lesions. While the initial impression was chronic graft-versus-host disease, additional work-up confirmed the diagnosis of DC. PMID- 15529129 TI - Primary small cell undifferentiated (neuroendocrine) carcinoma of the maxillary sinus. AB - Primary small cell undifferentiated (neuroendocrine) carcinomas of the paranasal sinuses are extremely uncommon neoplasms. This tumor was first reported in this site in 1965, and since then there have been only 61 documented cases in the literature. The median age at presentation is 53 years, with no gender predilection. There is no reported association of occurrence of this tumor with either tobacco use or form of occupation, and case outcome is usually poor. We report a case in a 25-year-old man, initially treated as an odontogenic infection and thus delaying institution of appropriate management. Further investigation identified a locally advanced neuroendocrine carcinoma of the left maxilla. Despite radiotherapy and chemotherapy, the patient exhibited rapid tumor dissemination and died. PMID- 15529130 TI - Clear cell adenocarcinoma of the tongue. AB - A firm, ulcerated tumor formed on the left side of the tongue of an elderly woman. Histopathological analysis showed that this unusual neoplasm was composed of monomorphic polygonal cells that exhibited a clear cytoplasm containing large amounts of periodic acid Schiff (PAS)-positive, diastase-digestive material. Most of the tumor cells stained immunohistochemically for Cytokeratin, high-molecular, CAM5.2, and epithelial membrane antigen (EMA), but were negative for alpha-smooth muscle actin, vimentin, glial fibrillary acid protein (GFAP), and S-100 protein. These findings supported a diagnosis of clear cell adenocarcinoma. Although patients with this type of tumor generally have a favorable prognosis, the tumor in our patient was fast-growing and contained a large number of Ki-67 positive cells, which are known to be highly proliferative. Thus, this case highlights the fact that even clear cell adenocarcinomas that are usually slow-growing should be investigated by conventional morphological techniques and their proliferative activity quantified in order to select the most appropriate treatment strategy. PMID- 15529131 TI - Adenoid ameloblastoma with dentinoid: a case report. AB - A man had a lesion of the anterior mandible that was initially diagnosed at 39 years of age as an adenomatoid odontogenic tumor. The lesion recurred 3 times over a span of 16 years. A consultative review of all histological findings was done and the tumor was reclassified as an ameloblastoma. PMID- 15529132 TI - Anatomical considerations for the spread of odontogenic infection originating from the pericoronitis of impacted mandibular third molar: computed tomographic analyses. AB - OBJECTIVES: The aims of the present study were to clarify the anatomy of impacted mandibular third molars in relation to surrounding structures and to investigate the pathway of infection originating from pericoronitis of this tooth. STUDY DESIGN: Computed tomography (CT) images were evaluated in 87 patients with uninfected mandibular third molar impaction and in 12 patients with infection originating from an impacted mandibular third molar. In uninfected patients, bony features around the impacted crown were investigated together with the relationship between the crown and surrounding muscles. In infected patients, involvements of bony and soft tissue structures were evaluated according to the disappearance of cortices and lateral asymmetry of density and shape in the spaces and muscles. RESULTS: In uninfected patients, the disappearance of the lingual cortical plate was observed in 48 (35.3%) impacted molars, while only in 11 (8.1%) teeth for buccal cortices. The cortical thickness was thinner on the lingual side than the buccal side. Sixty-five percent of the masseter muscle horizontally overlapped the crown, while almost all of the medial pterygoid muscle was posteriorly situated apart from the crown. The mylohyoid muscle horizontally overlapped the crown at below or intermediate vertical positions. In infected patients, the involvement of lingual structures was more frequently observed than that of buccal structures. The mylohyoid muscle was involved in 10 (83.3%) of 12 patients. Among them, 8 showed submandibular space involvement. CONCLUSION: CT findings supported the clinical observations of infection spread in patients with pericoronitis of the impacted mandibular third molar. CT appeared to be an effective tool for investigating the pathway of infection originating from the pericoronitis of impacted mandibular third molars. PMID- 15529133 TI - An evaluation of the origin of trabecular bone patterns using visual and digital image analysis. AB - OBJECTIVE: The purpose of this research project was to investigate the origin of the anatomical structures interpreted as trabecula bone on dental radiographic images. STUDY DESIGN: Mandible sections were cut sagitally into halves. Trabecular bone was removed from each section in 4 stages. Following each stage, standardized radiographs were made, using CDR direct digital equipment. Trabecular bone in the resulting digital images was measured with 4 methods: (1) mean gray level; (2) the fractal dimension of the basic images; and, following morphological image processing, (3) counting the number of trabecular ends, intercepts, and segments (EIS) and (4) performing fractal analyses of the skeletonized images. Additionally, human visual interpretation of the collected images was conducted through a written examination. Repeated measures analysis of variance (ANOVA) was used to test for changes in measurements attributable to bone removal. RESULTS: Repeated measures ANOVA indicated that the use of gray levels, fractal dimension, and morphologic operations quantifying using EIS or fractal analysis had similar performance and resulted in significant changes in measurements following bone removal ( P < .05). Visual differences were not always apparent between each stage of bone reduction. Radiometric and morphologic analysis showed measurable differences between stages. CONCLUSIONS: These results imply that the inner trabecula, the junctional trabecula, and the actual cortical housing all contribute to some extent to the radiograph, although changes in the radiographic architecture are not always clinically detectible. PMID- 15529134 TI - Factors associated with the removal of fractured NiTi instruments from root canal systems. AB - OBJECTIVE: To evaluate the influence of various factors on the success or failure of attempts to remove fragments of separated NiTi instruments from root canals. STUDY DESIGN: Instrument removal attempts were undertaken on 72 teeth with a separated NiTi instrument using a variety of techniques and armamentarium. Factors including type of tooth, degree of root canal curvature, location of fragment in relation to the root canal curvature, and radiographic length of fragment were analyzed. A success of treatment was defined as removal or complete bypassing of the fragments. RESULTS: The overall success rate was 53%. The success rate for ProFile fragments was 41% and for the NiTi K-file 60%. The success rate in maxillary teeth was higher than that in mandibular teeth. Of 52 instruments in molars, 28 were successfully removed or bypassed. Of the 12 fragments in premolars, only 2 were removed. All 8 cases in anterior teeth were retrieved completely. When the fragment was localized before the curvature, complete removal was achieved. When the fragments were located at and beyond the curvature, the success rates were 60% and 31%, respectively. In canals with a slight, moderate, and severe curvature, the success rates were 100%, 83%, and 43%, respectively. In general, the longer the fragment, the greater the chance for successful removal or bypass. CONCLUSIONS: Favorable factors for removal of separated NiTi fragments are straight root canals, anterior teeth, localization before the curvature, fragments longer than 5 mm, and hand NiTi K-file. PMID- 15529135 TI - Effects of ultrasonic root end preparation on resected root surfaces: SEM evaluation. AB - OBJECTIVE: The purpose of this study was to investigate the in vitro effect of ultrasonic retrotips on root end surfaces. STUDY DESIGN: Root end resection was performed on 45 single-root teeth endodontically treated after extraction. Setting the ultrasonic device at full power, a retrograde cavity was made by a stainless steel tip in 9 specimens (SS-FP). In another 9 samples a diamond tip was used (D-FP). Setting the intensity of the ultrasonic device at half power, 9 specimens were treated using stainless steel tips (SS-HP) and 9 using diamond tip (D-HP). Nine teeth were only apically resected and used as controls. Histologic serial sections were examined by scanning electron microsope to assess the number of root-face cracking, the marginal quality, and the crack type. RESULTS: No significant difference between diamond and stainless steel groups was found at a given power setting. Significant differences were found between SS-FP and SS-HP group for both the number of cracks and the marginal quality. PMID- 15529136 TI - Effects of cured dentin bonding materials on human monocyte viability. AB - OBJECTIVES: Dentin Bonding Agents (DBAs) have been proposed as root-end filling materials. This study examined the effect of polymerized DBAs on human monocyte viability. STUDY DESIGN: Monocytes were directly isolated from peripheral blood and being exposed to cured Scotch bond I (Single Bond) and Prime & Bond in different time intervals (36 and 72 hours). The viability of monocytes was determined by MTT assay. RESULTS: Viability of the cells was time dependent. There was no significant difference between the effect of 2 DBAs on monocytes. CONCLUSION: Results indicate that DBAs in polymerized form can alter the viability of monocytes and decrease it within time. PMID- 15529137 TI - Overdue process. PMID- 15529138 TI - Industry wary of NIH consultancy moratorium. PMID- 15529139 TI - Illegal seeds overtake India's cotton fields. PMID- 15529140 TI - US NCI launches nanotechnology plan. PMID- 15529141 TI - Kenyan dispute illuminates bioprospecting difficulties. PMID- 15529143 TI - Profile: Thomas Lonngren. PMID- 15529145 TI - Biogenerics standoff. PMID- 15529146 TI - The Cartagena protocol and GMOs. PMID- 15529147 TI - Ethics, industry and 'animal farm'. PMID- 15529149 TI - Monitoring horizontal gene transfer. PMID- 15529151 TI - Glycoprotein sialylation in plants? PMID- 15529153 TI - Changes in the business of culture. PMID- 15529154 TI - How similar do 'biosimilars' need to be? PMID- 15529155 TI - Quantum dots as photosensitizers? PMID- 15529156 TI - The best companies bare all. PMID- 15529157 TI - Biomanufacturing, from bust to boom...to bubble? PMID- 15529158 TI - Turning aptamers into anticoagulants. PMID- 15529159 TI - Fluorescent proteins in a new light. PMID- 15529160 TI - Finding time for allosteric interactions. PMID- 15529161 TI - DNA as a template for reaction discovery. PMID- 15529163 TI - Characterizing biological products and assessing comparability following manufacturing changes. AB - Changes in production methods of a biological product may necessitate an assessment of comparability to ensure that these manufacturing changes have not affected the safety, identity, purity, or efficacy of the product. Depending on the nature of the protein or the change, this assessment consists of a hierarchy of sequential tests in analytical testing, preclinical animal studies and clinical studies. Differences in analytical test results between pre- and post change products may require functional testing to establish the biological or clinical significance of the observed difference. An underlying principle of comparability is that under certain conditions, protein products may be considered comparable on the basis of analytical testing results alone. However, the ability to compare biological materials is solely dependent on the tests used, since no single analytical method is able to compare every aspect of protein structure or function. The advantages and disadvantages of any given method depends on the protein property being characterized. PMID- 15529164 TI - Production of recombinant protein therapeutics in cultivated mammalian cells. AB - Cultivated mammalian cells have become the dominant system for the production of recombinant proteins for clinical applications because of their capacity for proper protein folding, assembly and post-translational modification. Thus, the quality and efficacy of a protein can be superior when expressed in mammalian cells versus other hosts such as bacteria, plants and yeast. Recently, the productivity of mammalian cells cultivated in bioreactors has reached the gram per liter range in a number of cases, a more than 100-fold yield improvement over titers seen for similar processes in the mid-1980s. This increase in volumetric productivity has resulted mainly from improvements in media composition and process control. Opportunities still exist for improving mammalian cell systems through further advancements in production systems as well as through vector and host cell engineering. PMID- 15529165 TI - Recombinant protein folding and misfolding in Escherichia coli. AB - The past 20 years have seen enormous progress in the understanding of the mechanisms used by the enteric bacterium Escherichia coli to promote protein folding, support protein translocation and handle protein misfolding. Insights from these studies have been exploited to tackle the problems of inclusion body formation, proteolytic degradation and disulfide bond generation that have long impeded the production of complex heterologous proteins in a properly folded and biologically active form. The application of this information to industrial processes, together with emerging strategies for creating designer folding modulators and performing glycosylation all but guarantee that E. coli will remain an important host for the production of both commodity and high value added proteins. PMID- 15529166 TI - Advances in the production of human therapeutic proteins in yeasts and filamentous fungi. AB - Yeast and fungal protein expression systems are used for the production of many industrially relevant enzymes, and are widely used by the research community to produce proteins that cannot be actively expressed in Escherichia coli or require glycosylation for proper folding and biological activity. However, for the production of therapeutic glycoproteins intended for use in humans, yeasts have been less useful because of their inability to modify proteins with human glycosylation structures. Yeast glycosylation is of the high-mannose type, which confers a short in vivo half-life to the protein and may render it less efficacious or even immunogenic. Several ways of humanizing yeast-derived glycoproteins have been tried, including enzymatically modifying proteins in vitro and modulating host glycosylation pathways in vivo. Recent advances in the glycoengineering of yeasts and the expression of therapeutic glycoproteins in humanized yeasts have shown significant promise, and are challenging the current dominance of therapeutic protein production based on mammalian cell culture. PMID- 15529167 TI - Plant cell cultures for the production of recombinant proteins. AB - The use of whole plants for the synthesis of recombinant proteins has received a great deal of attention recently because of advantages in economy, scalability and safety compared with traditional microbial and mammalian production systems. However, production systems that use whole plants lack several of the intrinsic benefits of cultured cells, including the precise control over growth conditions, batch-to-batch product consistency, a high level of containment and the ability to produce recombinant proteins in compliance with good manufacturing practice. Plant cell cultures combine the merits of whole-plant systems with those of microbial and animal cell cultures, and already have an established track record for the production of valuable therapeutic secondary metabolites. Although no recombinant proteins have yet been produced commercially using plant cell cultures, there have been many proof-of-principle studies and several companies are investigating the commercial feasibility of such production systems. PMID- 15529168 TI - Publish and perish: what constitutes a bar under the patent laws. AB - In a recent decision, the Federal Circuit continues to expand the definition of a printed publication with regard to patentable matter. PMID- 15529172 TI - How do RNA folding algorithms work? AB - Programs such as MFOLD and ViennaRNA are widely used to predict RNA secondary structures. How do these algorithms work? Why can't they predict RNA pseudoknots? How accurate are they, and will they get better? PMID- 15529173 TI - A common open representation of mass spectrometry data and its application to proteomics research. AB - A broad range of mass spectrometers are used in mass spectrometry (MS)-based proteomics research. Each type of instrument possesses a unique design, data system and performance specifications, resulting in strengths and weaknesses for different types of experiments. Unfortunately, the native binary data formats produced by each type of mass spectrometer also differ and are usually proprietary. The diverse, nontransparent nature of the data structure complicates the integration of new instruments into preexisting infrastructure, impedes the analysis, exchange, comparison and publication of results from different experiments and laboratories, and prevents the bioinformatics community from accessing data sets required for software development. Here, we introduce the 'mzXML' format, an open, generic XML (extensible markup language) representation of MS data. We have also developed an accompanying suite of supporting programs. We expect that this format will facilitate data management, interpretation and dissemination in proteomics research. PMID- 15529174 TI - Promoter prediction analysis on the whole human genome. AB - Promoter prediction programs (PPPs) are important for in silico gene discovery without support from expressed sequence tag (EST)/cDNA/mRNA sequences, in the analysis of gene regulation and in genome annotation. Contrary to previous expectations, a comprehensive analysis of PPPs reveals that no program simultaneously achieves sensitivity and a positive predictive value >65%. PPP performances deduced from a limited number of chromosomes or smaller data sets do not hold when evaluated at the level of the whole genome, with serious inaccuracy of predictions for non-CpG-island-related promoters. Some PPPs even perform worse than, or close to, pure random guessing. PMID- 15529175 TI - Ask the experts: how to get hired. PMID- 15529176 TI - Perp is required for tissue-specific cell survival during zebrafish development. AB - The tumor suppressor p53 has two alternative effects, causing either cell cycle arrest or apoptosis. These different effects are supposed to be mediated by the transcriptional activation of different target genes. perp, encoding a transmembrane protein of the Pmp22 family, is a transcriptional p53 target exclusively upregulated in apoptotic cells. However, its role during normal development had remained largely unclear. Here, we report the isolation and characterization of a zebrafish perp homolog. Upon overexpression in early zebrafish embryos, perp induces apoptosis. In addition, it contributes to p53 dependent and UV-induced cell death. However, during normal zebrafish development, perp displays a p53-independent and spatially restricted expression in specific cell types and tissues. Antisense-mediated loss of Perp function leads to increased apoptosis in perp-expressing cells of the developing skin and notochord. We conclude that, in contrast to its proapoptotic function in stressed cells, Perp plays an antiapoptotic role during normal zebrafish development to regulate tissue-specific cell survival. PMID- 15529177 TI - Promyelocytic leukemia protein-induced growth suppression and cell death in liver cancer cells. AB - The promyelocytic leukemia protein (PML), involved in the pathogenesis of acute promyelocytic leukemia, is a coactivator of p53 tumor suppressive functions. The ability of PML to inhibit growth and induce cell death in solid tumor cells, however, has not been determined. We therefore assayed the tumor suppressor activities of PML and compared them with those of p53 in four liver cancer cell lines. Following infection of cells with replication-deficient recombinant PML adenovirus, the exogenous PML localized in the nucleus and formed abnormally enlarged PML-nuclear bodies after 24 hours. In vitro growth curve analysis showed that the overexpressed PML initially induced a substantial G1 cell cycle arrest and triggered massive cell death in all tested cell lines, irrespective of their p53 status. PML-induced cell death decreased by about 30% in the presence of a broad caspase inhibitor, zVAD. The cell death effect of PML was higher than that induced by p53 over a longer period of time. As with p53, overexpression of PML was closely related to upregulation of p21 and decrease of cyclin D1 expression. Unexpectedly, retinoic acid (RA) antagonized rather than enhanced PML-triggered cell death. RA enhanced the expression of adenovirus-cytomegalovirus-promoted PML at both transcription and protein levels within 12 hours after treatment; however, the PML protein was significantly degraded in the presence of RA at days 3-5 postinfection. PML degradation was also observed in SK-BR3 breast cancer cells treated with RA. Taken together, our findings strongly support the hypothesis that PML acts as a strong independent cell death inducer and that RA conversely abolishes the therapeutic effects of the PML proteins through proteasomal degradation of the protein. PMID- 15529178 TI - Pulmonary lymphohistiocytic reactions temporally related to etanercept therapy. AB - This report details the pulmonary pathologic findings in four patients with rheumatoid arthritis, who developed new onset of pulmonary signs and symptoms with alveolar infiltrates temporally related to the institution of etanercept therapy. Biopsy findings showed an interstitial and air space lymphohistiocytic infiltrate with non-necrotizing granulomas, in the setting of negative cultures and special stains for microorganisms. The association with etanercept therapy and granulomatous reactions is discussed along with the differential diagnosis. PMID- 15529179 TI - In vivo assessment of melanocytic nests in nevi and melanomas by reflectance confocal microscopy. AB - In vivo reflectance confocal microscopy is a novel technique for the noninvasive study and diagnosis of the skin. The aim of this study was to describe and characterize the cytological and architectural aspects of cell clusters in melanocytic lesions observed by confocal microscopy, and to correlate them with routine histopathology. A total of 55 melanocytic lesions comprising 20 melanomas, 25 acquired nevi and 10 Spitz nevi were studied by means of reflectance confocal microscopy, dermoscopy and routine histopathology. Three different types of cell clusters at confocal microscopy observation (dense, sparse cell and cerebriform clusters) were identified and correlated with histopathology. Dense clusters appeared characteristic for benign lesions, although present in 13 out of 20 melanomas. Sparse cell clusters were more frequently observable in melanomas, but also sporadically present in one Spitz nevus. Moreover, cerebriform clusters were exclusively observed in five out of 20 melanomas. Confocal microscopy allowed the in vivo characterization of aspects of melanocytic nests and their exact correlation with histopathology. PMID- 15529180 TI - p63 Immunohistochemistry in the distinction of adenoid cystic carcinoma from basaloid squamous cell carcinoma. AB - Morphologic distinction of high-grade adenoid cystic carcinoma from basaloid squamous cell carcinoma can be difficult. Equivocal diagnoses can mislead treatment. We have investigated the possibility that immunohistochemical staining for the presence of p63, a novel epithelial stem-cell regulatory protein, could be a useful means of distinguishing these two neoplasms. Archival, routinely processed slides were subjected to citrate-based antigen retrieval, exposure to anti-p63 monoclonal 4A4, and developed with a streptavidin-biotin kit and diaminobenzidine as chromogen. p63 was detected in 100% of the adenoid cystic carcinomas (n=14) and 100% of basaloid squamous cell carcinomas (n=16). Basaloid squamous cell carcinomas consistently displayed diffuse p63 positivity, with staining of nearly 100% of tumor cells. In contrast, adenoid cystic carcinoma displayed a consistently compartmentalized pattern within tumor nests. Compartmentalization was manifested in two patterns: (1) selective staining of a single peripheral layer of p63-positive cells surrounding centrally located tumor cells that were p63-negative and (2) tumor nests consisting of multiple contiguous glandular/cribriform-like units of p63-positive cells surrounding or interspersed with p63-negative cells. p63 immunostaining constitutes a specific and accurate means of distinguishing adenoid cystic carcinoma from basaloid squamous cell carcinoma. p63 positivity in adenoid cystic carcinoma appears to be homologous to that seen in the basal and/or myoepithelial compartments of salivary gland and other epithelia, and may signify a stem-cell-like role for these peripheral cells. Diffuse p63 positivity in basaloid squamous cell carcinoma suggests dysregulation of p63-positive stem cells in poorly differentiated squamous carcinoma. PMID- 15529181 TI - Prediction of endometrial carcinoma by subjective endometrial intraepithelial neoplasia diagnosis. AB - Endometrial intraepithelial neoplasia (also known as 'EIN') is a precursor to endometrioid endometrial adenocarcinoma characterized by monoclonal growth of mutated cells, a distinctive histopathologic appearance, and 45-fold elevated cancer risk. We have applied diagnostic criteria for EIN to 97 successive endometrial biopsies classified as hyperplastic according to World Health Organization criteria and correlated results with computer-assisted morphometry (D-score) and clinical cancer outcomes. Three pathologists separately reviewed all cases for presence or absence of EIN using published criteria (gland area>stromal area, cytologic change in focus of altered architecture, lesion size>1 mm, and exclusion of cancer and mimics). Discordant cases were resolved by a consensus review at a multiheaded scope. Clinical outcomes were obtained in 84 patients from patient visit and pathology records. Diagnoses of presence or absence of EIN were unanimous among all three pathologists in 75% of cases, and intraobserver-reproducibility was very good (kappa 0.73-0.90). Cases rediagnosed as EIN encompassed hyperplasias previously diagnosed as atypical (n=18) or nonatypical (eight complex, two simple). Eight follow-up cancers were scattered between hyperplasia types (5/21 atypical, 3/63 nonatypical), but all classified as EIN (8/25) and D-score or=50% of neoplastic cells showed immunostaining. In reactive lymph nodes, hENT1 was confined to the germinal centers, whereas mantle zone B-cells and interfollicular T-cells were negative. In NHL, a relatively high frequency of hENT1 positivity was found in Burkitt lymphoma/leukemia (63%), diffuse large B-cell lymphoma (DLCL; 45%), and follicular lymphoma (40%). In DLCL, 26% of cases were positive for CD10, and CD10 positive DLCL cases were more likely to be hENT1 positive than CD10-negative cases (P=0.025). A lower frequency of hENT1 positivity was found in mantle cell lymphoma (13%) and peripheral T-cell lymphomas (37%). All marginal zone lymphomas (n=5), chronic lymphocytic leukemia small lymphocytic lymphomas (n=10), plasmacytoma (n=3), acute lymphoblastic lymphoma/leukemia, and anaplastic large cell lymphomas (n=5) were negative. In conclusion, hENT1 was most frequently found in benign and malignant follicular center cells. Prospective studies to assess the value of hENT1 immunostaining in predicting resistance to nucleoside chemotherapy for NHL are warranted. PMID- 15529185 TI - Tumor classification by tissue microarray profiling: random forest clustering applied to renal cell carcinoma. AB - We describe a novel strategy (random forest clustering) for tumor profiling based on tissue microarray data. Random forest clustering is attractive for tissue microarray and other immunohistochemistry data since it handles highly skewed tumor marker expressions well and weighs the contribution of each marker according to its relatedness with other tumor markers. This is the first tumor class discovery analysis of renal cell carcinoma patients based on protein expression profiles. The tissue array data contained at least three tumor samples from each of 366 renal cell carcinoma patients. The eight tumor markers explore tumor proliferation, cell cycle abnormalities, cell mobility, and the hypoxia pathway. Since the procedure is unsupervised, no clinicopathological data or traditional classifications are used a priori. To explore whether the tissue microarray data can be used to identify fundamental subtypes of renal cell carcinoma patients, we first carried out random forest clustering of all 366 patients. By analyzing the tumor markers simultaneously, the procedure automatically detected classes that correspond to clear- vs non-clear cell tumors (demonstration of proof-of-principle). The resulting molecular grouping provides better prediction of survival (logrank P=0.000090) than this classical pathological grouping (logrank P=0.023). We then sought to extend the class discovery by searching for finer subclasses of clear cell patients. The procedure automatically discovered: (a) two classes corresponding to low- and high-grade patients (demonstration of proof-of-principle); (b) a subgroup of long-surviving clear cell patients with a distinct molecular profile and (c) two novel tumor subclasses in low-grade clear cell patients that could not be explained by any clinicopathological variables (demonstration of discovery). PMID- 15529187 TI - A brief history of the pathology of the gonads. AB - Our understanding of gonadal pathology has reached its current state as a result of the contributions of numerous outstanding investigators. Knowledge of testicular tumor pathology dates back to the great British workers Percival Pott and Sir Astley Cooper but the single greatest early stride was made with the description in 1906 by the French urologist Maurice Chevassu of the seminoma. The seminal 1946 paper of Nathan B Friedman and Robert A Moore, which segregated out as a distinct entity embryonal carcinoma, is, however, the foundation for the current classification of testicular tumors. In that year Pierre Masson described the distinctive neoplasm, the spermatocytic seminoma. The 1950s saw the publication of an important paper by Frank J Dixon and Dr Moore and they also wrote the first series fascicle on testicular tumors. In this same timeframe, and thereafter, Robert E Scully made significant contributions to testicular pathology, writing the first English language paper on spermatocytic seminoma, describing several subtypes of sex cord tumor, and also the distinctive lesion of intersex, the gonadoblastoma, as well as playing a major role in 1980 in formulating the current classification of premalignant lesions of the testis. The current classification of testicular tumors was arrived at in the early 1970s when the World Health Organization, under the leadership of Dr FK Mostofi, who himself made notable contributions to testicular pathology, devised what is fundamentally the current classification of neoplasms of the male gonad. Although comments on ovarian pathology were made by such legendary figures of earlier times as Giovanni Battista Morgagni and Matthew Baillie, it is only in the mid to later years of the 19th century that contributions, mostly in Europe, began to move knowledge of ovarian pathology to its current state. Thomas Hodgkin, Richard Bright, and Sir James Paget all wrote extensively on ovarian neoplasms. In 1870, Heinrich Waldeyer, and later in that century, another German, Hermann Johannes Pfannenstiel wrote important papers on the surface epithelial tumors. The latter was likely the first to refer to neoplasms now known as of 'borderline malignancy' and also wrote on pseudomyxoma peritonei and other topics. Their work was followed by that of Robert Meyer who made monumental contributions to gynecological pathology, including recognizing the Brenner tumor as a distinctive neoplasm and proposing the first classification of Sertoli-Leydig cell tumors (arrhenoblastomas). He also coined the term 'disgerminoma' (soon changed to dysgerminoma) for the ovarian tumor that had been described in detail by the French investigator Marcel Chenot 5 years after Chevassu had mentioned the tumor in his paper describing the seminoma. During the Meyer era other significant contributions were made by, among others, Howard C Taylor writing on the borderline tumors and John A Sampson writing on endometriosis and tumors, associated with it. In the second-half of the 20th century major contributions were made by Gunnar Teilum of Denmark and Lars Santesson of Sweden. Dr Teilum delineated the morphologic features of the yolk sac tumor and noted the resemblance of papillary formations within it to the endodermal sinuses of the rat placenta. He also wrote extensively on sex cord tumors in both gonads. At a FIGO meeting in 1961 Dr Santesson played a major role in formulating the first organized classification of the surface epithelial-stromal tumors of the ovary and also promoted the endometrioid carcinoma as a special variant of ovarian cancer. In a career spanning over 50 years, Dr Scully was the architect of the modern classification of ovarian tumors being the driving force behind the influential 1973 World Health Organization classification of them. His many original observations have touched upon virtually all categories of ovarian tumor pathology. His second series fascicle 'Tumors of the Ovaries and Maldeveloped Gonads' utilized the WHO classification and presented a lucid elaboration of his by then vast experience with ovarian tumors. All the above have left a rich legacy which those who follow in their path will be challenged to equal. PMID- 15529186 TI - Galectin-3 and HBME-1 expression in well-differentiated thyroid tumors with follicular architecture of uncertain malignant potential. AB - Well-differentiated encapsulated tumors of the thyroid gland with a follicular architecture may cause diagnostic difficulties. Questionable vascular or capsular penetration may raise the possibility of a follicular carcinoma, while focal nuclear clearing and grooves may suggest a diagnosis of papillary carcinoma. A proposal has recently been made to designate cases showing suggestive but not conclusive morphological evidence of malignancy along these lines as well differentiated or follicular tumors of uncertain malignant potential. The aim of the present study was to investigate the expression and diagnostic role in well differentiated or follicular tumors of uncertain malignant potential of Galectin 3 and HBME-1, two malignancy-related markers. A total of 21 tumors fulfilling the criteria of well-differentiated or follicular tumors of uncertain malignant potential were collected from two institutions, including eight cases with questionable vascular and/or capsular invasion and 13 cases with some degree of nuclear changes in the form of clearing, grooves, and/or pseudoinclusions. Tumors in the first group expressed HBME-1 and Galectin-3 focally (less than 25% of tumor cells) in 5/8 and 3/8 cases, respectively, with 62.5% of cases reacting for at least one marker. Cases in the second category expressed HBME-1 and Galectin-3 in 9/13 and 10/13 cases, respectively, with 92.3% of cases having at least one marker expressed. These findings indicate that HBME-1 and Galectin-3 are heterogeneously distributed in these borderline tumors, but that a strong and diffuse expression of HBME-1 and to a lower extent of Galectin-3 was preferentially observed in the group characterized by nuclear changes which were similar but less developed than those of conventional papillary carcinoma. The relationship found between the markers investigated and these nuclear changes suggests that the tumors containing them are pathogenetically linked to papillary carcinomas. PMID- 15529188 TI - Carbon dioxide inhibits the growth rate of Staphylococcus aureus at body temperature. AB - BACKGROUND: Since the 1930s, carbon dioxide (CO(2)) has been combined with cold storage for the preservation of food. However, its use for the prevention of surgical wound infection was long considered to be impractical. Now CO(2) is widely used during laparoscopic procedures, and a method has been developed to create a CO(2) atmosphere in an open wound. The aim of this study was to investigate the effect of CO(2) on the growth of Staphylococcus aureus at body temperature. METHODS: First, S. aureus inoculated on blood agar were exposed to pure CO(2) (100%), standard anaerobic gas (5% CO(2), 10% hydrogen, 85% nitrogen), or air at 37 degrees C for a period of 24 h; then a viable count of the bacteria was made. Second, S. aureus inoculated in brain-heart infusion broth and kept at 37 degrees C were exposed to CO(2) or air for 0, 2, 4, 6, and 8 h; then the optical density of the bacteria was measured. RESULTS: After 24 h, the number of S. aureus on blood agar was about 100 times lower in CO(2) than in anaerobic gas (p = 0.001) and about 1,000 times lower than in air (p = 0.001). Also, in broth, there were fewer bacteria with CO(2) than with air (p < 0.01). After 2 h, the number of bacteria was increased with air (p < 0.001) but not with CO(2) (p = 0.13). After 8 h, the optical density had increased from zero to 1.2 with air but it had increased only to 0.01 with CO(2) (p = 0.001). CONCLUSION: Pure CO(2) significantly decreased the growth rate of S. aureus at body temperature. The inhibitory effect of CO(2) increased exponentially with time. Its bacteriostatic effect may help to explain the low infection rates in patients who undergo laparoscopic procedures. PMID- 15529189 TI - Long-term outcome, adhesions, and quality of life after laparoscopic and open surgical therapies for acute abdomen: follow-up of a prospective trial. AB - BACKGROUND: The objective of this study was to determine the long-term outcome of laparoscopic and open therapies for acute abdomen, and to assess the patients postoperative quality of life, with special attention to adhesions. METHODS: A follow-up study was conducted from June through December 2001 of a case-control trial of laparoscopic and open surgical treatment in patients with acute abdomen. Mean (median) duration of follow-up was 35.9 months (32.5) for the laparoscopic group (L) and 40.7 months (38.5) for the open group (O). A total of 284 (92%) of the original 310 study patients (108 L and 202 O) were contacted. Twenty-eight (9%) were confirmed to be dead. From the remaining 256 patients (131 men and 125 women, mean age [+/- SD] 38.9 +/- 19.9 years), we enrolled 153(67 L and 91 O) in the follow-up. The main outcome measures were frequency of relapse requiring treatment, frequency of reoperations, incidence of adhesion ileus and incisional hernia, distant morbidity and mortality, satisfaction with therapy, and Gastrointestinal Quality of Life Index (GQLI) scores. RESULTS: The morbidity and mortality rates, readmission rates, incisional hernia rates, and scores for long term quality of life were the same in both groups. The reoperation rate was significantly higher among L patients, but there were significantly fewer episodes of adhesion ileus in this group. Patient satisfaction was 85% in the L group and 73% in the O group (p = NS). CONCLUSIONS: The laparoscopic treatment of patients with acute abdomen offers an outcome comparable to that achieved with the open approach. There were fewer episodes of adhesion ileus in laparoscopic patients. Consequently, the operative treatment of acute abdomen patients by laparoscopy can be recommended. PMID- 15529190 TI - Radiofrequency tissue ablation in an experimental model of grade IV renal trauma: a preliminary report. AB - BACKGROUND: The kidney is one of the most frequently injured intraabdominal organs. In this study, we investigated the efficacy and safety of radiofrequency ablation (RFA) as a kidney-preserving hemostatic technique for grade IV renal trauma. METHODS: A grade IV injury was induced in the right kidney of 12 Landrace pigs. Then RFA was applied around the injury in 10 animals until hemostasis was achieved; two animals were not treated (control group). The treated animals were killed humanely on days 0, 3, 7, 14 and 21 and examined. The kidneys were subjected to histologic and radiologic examination. RESULTS: The two untreated animals died from hypovolemic shock. Hemostasis was achieved in all treated animals. We had no operative deaths and no morbidity. No blood, pus, urine, or other fluid was found at the time of death. In one animal, a fistulous lesion leading to the collecting system was identified, but no urine leakage was observed. At histology, the ablated areas were found to consist of three zones: an inner necrotic one, a zone of neutrophils digesting necrotic tissue, and an outer zone with possibly reversible damage. CONCLUSIONS: Radiofrequency ablation is an efficient and safe hemostatic method for grade IV renal trauma. Further study is needed to investigate the possible application of this method to humans and its percutaneous or laparoscopic use. PMID- 15529191 TI - Power spectral analysis of heart rate variability during helium pneumoperitoneum: The mechanism of increased cardiac sympathetic activity and its clinical significance. AB - BACKGROUND: Carbon dioxide pneumoperitoneum (PP) is known to induce increased cardiac sympathetic expression. The role of the insufflated gas involved in this mechanism should be elucidated in an attempt to eliminate its possible serious consequences. METHODS: Twenty-five patients undergoing elective laparoscopic cholecystectomy were prospectively analyzed for cardiac autonomic nervous activity by spectral heart rate variability. In 15 patients, helium was used as CO(2) substitution for abdominal insufflation (study group). Four frequency bands of interest were obtained from the power spectrum of R-R intervals, as well as the ratio between the low and high frequency (LF/HF), using the fast Fourier transformation algorithm to characterize the synergy of both autonomic branches during PP. RESULTS: Significantly increased values of the power spectrum related to the LF and VLF bands (from 130 to 377 msec(2)/Hz and from 145 to 516 msec(2)/Hz, respectively) were inspected during CO(2) PP, as well as increased LF/LH ratio (2.1). Using helium as CO(2) substitution has eliminated the significant changes in the power spectrum that reflect increased cardiac sympathetic activity. CONCLUSIONS: The elimination of sympathetic predominance by helium PP indicates the central role of CO(2) in establishing this phenomena. Considering this information and its other known advantages, helium should be considered for use during prolonged laparoscopic procedures for high-risk patients. PMID- 15529192 TI - A new laparoscopic technique for the placement of a permanent peritoneal dialysis catheter: the preperitoneal tunneling method. AB - BACKGROUND: Mechanical outflow obstruction and leakage from the exit site of the catheter are two common complications of continuous ambulatory peritoneal dialysis. To lessen these complications and to achieve immediate use of the catheter, we developed a new laparoscopic technique for catheter placement. METHODS: A total of 12 consecutive patients with end-stage renal failure were included in this study between April 2003 and July 2003. The average age of the patients was 42.4 years (range, 37-72). Patients were excluded only if a serious risk for general anesthesia was found. Using two 5-mm ports and a 3.3-mm mini laparoscope, a peritoneal dialysis catheter was passed through a preperitoneal tunnel before the tip of the catheter was introduced into the pelvis. Routine peritoneal dialysis was started immediately after the operation while the patients were still in the operating room. RESULTS: The mean operating time was 18.6 min (range, 12-37). There was no operative morbidity. The mean follow-up period was 4.3 months (range, 3-7). No leakage of the dialysate liquid or outflow obstruction was observed during this period. CONCLUSION: The advantages of this method include accurate placement, preperitoneal fixation, and immediate use of the catheter for routine peritoneal dialysis. We also believe that because of the preperitoneal fixation of the catheter, this technique will decrease outflow obstruction, which usually occurs due to omental wrapping or displacement of the catheter tip. PMID- 15529193 TI - Influence of different gases used for laparoscopy (helium, carbon dioxide, room air, and xenon) on tumor volume, histomorphology, and leukocyte-tumor-endothelium interaction in intravital microscopy. AB - BACKGROUND: Previous studies indicate that helium pneumoperitoneum used for laparoscopic surgery suppresses whereas carbon dioxide pneumoperitoneum increases postoperative tumor growth. The pathomechanisms of decreased tumor growth by helium are unknown. This study was designed to examine the effect of the gases helium, carbon dioxide (CO(2)), and air, and xenon, which can be used to induce pneumoperitoneum in laparoscopy on tumor volume, histomorphology, and leukocyte endothelium interaction measured by intravital microscopy in rats with implanted liver malignoma (Morris hepatoma 3924A). METHODS: In 46 rats, Morris hepatoma 3294A cells were implanted intrahepatically. After implantation, rats were randomized into two main groups. In the first main group, 10 animals were prepared for examination of leukocyte-endothelium interaction by intravital video microscopy and were randomized into two groups. Five days after implantation they underwent laparoscopy using either helium (n = 5) or CO(2) (n = 5). Ten days after implantation the rats underwent intravital video microscopy to assess leukocyte-endothelium interaction in the tumor and liver vessels. In the second main group 36 rats were prepared for examination of tumor volume arid histomorphology. They were randomized into five groups. Five days after implantation they underwent laparoscopy using helium (n = 7), carbon dioxide (n = 7), room air (n = 7), or xenon (n = 8). The control group (n = 7) received anesthesia only. Rats were killed 10 days after tumor implantation to assess tumor volume and histomorphology. RESULTS: Compared to the control group or groups that received CO(2), room air, or xenon for pneumoperitoneum, the establishment of helium pneumoperitoneum caused a significantly smaller tumor volume (Kruskal-Wallis test, p = 0.001; median tumor-volume: control group, 44 mm(3); helium 19 mm(3)). There was no significant difference in histomorphology between the groups. There was only a statistically significant difference in the development of central tumor necrosis in accordance to tumor volume (Mann-Whitney test, p = 0.03). In the tumor samples, roller counts were statistically significantly higher in the helium group compared to the CO(2) group (p = 0.04). For sticker counts, no statistically significant effects due to liver/tumor (p = 0.13) or treatment (p = 0.48) were observed. CONCLUSIONS: There was a significant decrease in tumor volume using helium pneumoperitoneum for laparoscopy compared to the other gases. Here, we demonstrate that suppression of tumor growth is not due to variation of histomorphology. It seems that helium pneumoperitoneum effects a higher leukocyte-endothelium interaction and thereby a higher immune activation. This could be one explanation for the statistically significantly smaller tumor volume after laparoscopy with helium compared to laparoscopy with CO(2). PMID- 15529194 TI - Peritoneal pH during laparoscopy is dependent on ambient gas environment: helium and nitrous oxide do not cause peritoneal acidosis. AB - BACKGROUND: Little is know about the effects of different insufflation gases on peritoneal pH during laparoscopy. However, these changes may influence the intracellular signalling system, resulting in altered cell growth or adhesiveness. The aim of this study was to determine the effects of carbon dioxide (CO(2)), nitrous oxide (N(2)O), and helium (He) on parietal and visceral peritoneal pH. The effect of different intraabdominal pressures on parietal and visceral peritoneal pH was also examined. METHODS: We conducted both an ambient gas study and a pressure study. For the ambient gas study, 20 pigs were divided into the following four groups: (a) CO(2), (b) He, (c) N(2)O, and (d) abdominal wall lift (Lift) laparoscopy. Parietal and visceral peritoneal pH were measured at 15 min intervals for 180 min. For the pressure study, 15 pigs were divided into the following three groups: (a) CO(2), (b) He, (c) N(2)O laparoscopy. Baseline values were established for parietal and visceral peritoneal pH. Intraabdominal pressure was then increased stepwise at 1-mmHg intervals to 15 mmHg. After pressure was maintained for 15 min at each setting, parietal and visceral peritoneal pH were measured. RESULTS: Ambient gas environment was the major determinant of parietal peritoneal pH. Carbon dioxide caused parietal peritoneal acidosis. Helium, N(2)O, and Lift caused alkalotic parietal peritoneal pH. Intraabdominal pressure had a minor effect on parietal peritoneal pH. At higher intraabdominal pressure (12-15 vs 5-8 mmHg), CO(2) caused a slight decrease in parietal peritoneal pH, whereas N(2)O and He caused a slight increase in parietal peritoneal pH. Visceral peritoneal pH remained relatively unaffected during all studies. CONCLUSIONS: Parietal peritoneal pH during laparoscopy was highly dependent on the ambient gas environment. The effect of intraabdominal pressure on parietal peritoneal pH was of minor significance. Carbon dioxide caused a slight worsening of parietal peritoneal acidosis at higher intraabdominal pressure, whereas, N(2)O, He, and Lift did not cause parietal peritoneal acidosis. PMID- 15529196 TI - Symptomatic internal hernias after laparoscopic bariatric surgery. AB - BACKGROUND: The aim of this study was to describe the occurrence and clinical characteristics of symptomatic internal hernias (IH) after laparoscopic bariatric procedures. METHODS: We conducted a retrospective review of cases of IH after 1,064 laparoscopic gastric bypasses (LGB) and biliopancreatic diversions with duodenal switch (LBPD-DS) performed from September 1998 to August 2002. RESULTS: We documented 35 cases of IH (overall incidence of 3.3%). The IH occurred in 6.0% of patients with retrocolic procedures and 3.3% of patients with antecolic procedures. Most were in the Petersen defect (55.9%) and at the enteroenterostomy site (35.3%). A bimodal presentation was observed, with 22.9% of patients with IH diagnosed in the early postoperative period (2-58 days) and 77.1% in a delayed fashion (187-1,109 days). A laparoscopic approach to the repair of IH was possible in 60.0% of patients. Complications occurred in 18.8% of patients, including one death (2.9%). CONCLUSION: Complete closure of all mesenteric defects is strongly recommended during laparoscopic bariatric procedures to avoid IH and their associated complications. PMID- 15529197 TI - Laparoscopic palliative surgery for complicated colorectal cancer. AB - BACKGROUND: The aim of this study was to evaluate the feasibility and outcomes of the laparoscopic approach for the palliation of advanced complicated colorectal cancer (CRC). METHODS: We reviewed 21 laparoscopic palliative procedures for emergent complications of advanced CRC between 1994 and 2002. Intraoperative complications, estimated blood loss, transfusions, operative times, time to first bowel movement, length of hospital stay, and postoperative complications were assessed. RESULTS: Indications for surgery included perforation (n = 10), bleeding (n = 7), and obstruction (n = 4). A proximal diverting procedure was performed in all patients, and a concomitant colon resection was performed in 18 patients (86%). The mean operative time was 181 +/- 22 min. Estimated blood loss was 283 +/- 48 cc, with three patients (14%) requiring transfusions. The average length of hospital stay was 8.6 +/- 2 days, and time to first bowel movement was 61 +/- 9 h. The complication rate and the 30-day mortality rate were 33% and 0%, respectively. CONCLUSION: A laparoscopic approach to address advanced CRC is safe and effective and should be considered part of the surgeon's armamentarium for the palliation of advanced complicated CRC. PMID- 15529199 TI - Altering first impressions after facial plastic surgery. AB - BACKGROUND: First impressions are greatly influenced by facial appearance. Clinical data from psychiatric medicine overwhelming indicate that attractive individuals receive better judgment, treatment and behavior [1,2,10]. This study aimed to determine whether cosmetic alteration of facial features projects a better first impression. METHODS: Random reviewers were asked independently to grade standardized preoperative and postoperative photographs of patients who underwent facial plastic surgery. The reviewers were blinded to the pre- or postoperative status of the photograph. The questions posed to the reviewers were based on first-impression studies used in the past. RESULTS: The findings indicate that postoperative cosmetic surgery patients were graded as 31% more attractive, 27% better in social skills, 22% more successful in dating, 19% better in athletic skills, 15% better in relationship skills, and 13% more financially successful. CONCLUSION: Facial cosmetic surgery can improve the first impression an individual creates. PMID- 15529201 TI - Breast reduction with no vertical scar. PMID- 15529202 TI - Rotation of anatomic prostheses: a possible cause of breast deformity. PMID- 15529203 TI - The minimal approach: an innovation in facial cosmetic procedures. AB - Plastic surgery is a rapidly growing field, particularly the use of less invasive procedures such as biodegradable dermal fillers and botulinum toxin. Aging is a complex process involving two important factors: volume loss throughout the face, and repetitive muscle movements that cause wrinkles and folds. Dermal fillers work by providing support for facial structures, whereas botulinum toxin reduces the mimetic effects. In combination, these products can be used effectively to reshape and rejuvenate the face and neck. Dermal fillers can be used throughout the face to raise the eyebrows, fill the tear trough, reshape the nose, lift the nose tip, fill nasolabial folds and oral comissures, fill the cheeks, raise the cheekbones, reshape the jaw line, and rejuvenate the neck area. This "minimal approach" offers a faster, less painful, and less costly alternative or complement to surgical facelifts. A variety of biodegradable injectable products are currently available, the most common of which are collagen- or hyaluronic acid-based. Using these products in combination takes advantage of their hydrophobic and hydrophilic properties and can provide better and longer-lasting results. The addition of botulinum toxin can further extend the duration of results by reducing the mimetic stress that can break down the polymers. With regular maintenance treatments, the minimal approach using dermal fillers and botulinum toxin can give patients a "whole new look" with a fast and relatively painless lunchtime procedure. PMID- 15529204 TI - One-stage augmentation combined with mastopexy: aesthetic results and patient satisfaction. AB - Since the original descriptions by Gonzales-Ulloa in 1960 and Regnault in 1966, breast augmentation in combination with mastopexy has remained a difficult, and often polarizing, topic in plastic surgery, not only because of its results but also because of its litany of potential complications. Over the past few years, there has been an increase in the discussion of one-stage augmentation combined with mastopexy throughout the literature. However, a critical analysis of the aesthetic results, as well as patient satisfaction with the procedure, continues to be absent. Because there have not been any reported studies on the aesthetic results or patient satisfaction with augmentation and mastopexy, we undertook this retrospective review in an attempt answer a fundamental question: is one stage breast augmentation combined with mastopexy aesthetically and functionally worthwhile for both the physician and patient? All 34 patients reviewed for this retrospective study underwent bilateral, one-stage breast augmentation and mastopexy between April 1996 and December 2002. Patient charts were reviewed for a number of parameters including previous breast surgery, degree of preoperative ptosis, type of mastopexy used, size and type of implants placed, implant position, postoperative complications, and any revision surgeries performed. Patient photographs were evaluated by observers blinded to the study, and patients were asked to complete a satisfaction questionnaire. Ptosis was graded according to the Regnault classification. As a result, 14 women had grade 1 ptosis (41%), fourteen had grade 2 ptosis (41%), one had grade 3 ptosis (3%), two had pseudoptosis (6%), and two had tuberous breasts (6%). The grade of ptosis in one patient was not defined. The patient complication rate was 8.8% (3 patients). For the aesthetic rating scale, preoperative and postoperative photographs taken after more than 1 year were evaluated. On the scale of 1 (poor) to 4 (excellent), overall ptosis correction was rated as 3.4, asymmetry correction as 3.4, postoperative breast symmetry as 3.2, scar quality as 3.3, breast shape as 3.1, nipple/areola size as 2.9, and overall result as 3. Only 13 of the 34 patients were available for completion of the satisfaction survey. Evaluation of the 13 patient satisfaction surveys showed that, on the average, the patients were satisfied with the various aspects of their surgery. The average overall result and surgical goals both were 3.1. However, 54% of the patients (n = 7) desired revision surgery for various reasons, the most common being a desire for more breast lift. A review of the patients and results, brought a number of issues to light. First, aesthetic results for augmentation and mastopexy truly depend on a number of different factors that must work in harmony to yield an excellent result. Second, what is aesthetically pleasing to the surgeon may not be pleasing to the patient, and vice versa. Third, although the patient aesthetic results were good, they were not consistently rated as excellent, nor were the patients totally satisfied with their outcomes. This perhaps reflects the more complex nature of both the patient's problems and the surgical procedure itself. Finally, although the overall results of one-stage breast augmentation and mastopexy are good, and the patients generally are satisfied, this study raises the question whether staging the surgery by performing the mastopexy first may not yield significantly better results than the combined simultaneous procedure. PMID- 15529205 TI - Estimation of breast prosthesis volume by the Cavalieri principle using magnetic resonance images. AB - BACKGROUND: The purpose of this study was to search the accuracy and reproducibility of the Cavalieri method for the estimation of the breast prosthesis volume on magnetic resonance images (MRI). METHODS: Nine breast prostheses were scanned in axial and coronal section planes by MRI. The consecutive sections in 5-mm thickness were taken, after which the volumes of the prostheses were measured and estimated by means of the fluid displacement technique and three-dimensional postprocessing software, respectively. The volumes of the prostheses also were estimated by three performers using the Cavalieri principle to check the accuracy and interobserver interrelation of the suggested technique RESULTS: There were no significant differences between the Cavalieri estimate and the real volumes of the breast prostheses (p > 0.05), nor between the performers's volume estimates (p > 0.05). The orientation of the section planes did not affect the accuracy of the estimates (p > 0.05) CONCLUSIONS: This report describes a rapid, simple, accurate, and practical technique for estimating the volume of the breast prostheses without changing the routine process in the clinics. It was concluded that the combination of MRI with the Cavalieri principle is a direct and accurate technique that can be applied safely for estimating the volume of the breast prosthesis in 4 min and 34 s per prosthesis. PMID- 15529207 TI - Neodymium: yttrium-aluminum-garnet long impulse laser for the elimination of superfluous hair: experiences and considerations from 3 years of activity. AB - This study examined the results obtained with a modern apparatus for laser hair removal (neodymium: yttrium-aluminum-garnet [Nd:YAG] laser at long impulses with a wave-length of 1,064 nm; Q-switched laser) over a follow-up period of 3 years. A large heterogeneous group of 480 patients was taken into consideration. These patients were treated according to a standard protocol with monthly checkups and a personalized protocol at deferred appointments. The results, discovered by means of the most objective procedure possible, were retrieved and put into a graph showing two different curves for the repopulation of hair. In their clinical travels, the authors observed an average variable regrowth of 40% to 65%, allowing them to affirm that laser hair removal using Nd:YAG at long impulses is decisively efficient in obtaining long-term results. The use of a protocol (denominated "prolonged monthly checkup") with laser sessions at ever decreasing periods permits, among other things, more outstanding and advantageous results for the patient. Thanks to more efficiently synchronized phases of the biologic hair cycle, this shortens and moves the telegenic phases closer and also renders the anagenic phases (those in which the selective photoermolysis on the pilipheric follicle proves to be efficient) more efficient. Personalization of the treatment relative to the monthly health checkup sessions is of fundamental importance to the scope of obtaining the best results in terms of cost-benefit rate, provided submassimal fluxes are (i.e., those well-tolerated by the patient) used. All this allows hair removal that is not definitive, but which becomes progressively permanent (i.e., characterized by ever-growing periods of lack of hair sustained by sporadic maintenance laser sessions based on the individual's necessity). PMID- 15529209 TI - [Comparative analysis of attributional style and self-esteem in a sample of depressed patients and normal control subjects]. AB - INTRODUCTION: The present study assesses the role of attributions and self-esteem in depressive states. In depression, attributions can become central biased coping mechanisms that guide individual behavior. METHODS: Self-esteem and attribution bias was assessed in 40 depressed and 80 normal controls. RESULTS: Results of this investigation show that depressed patients made more internal attributions to negative events, and more external attributions for positive events than their normal counterparts, confirming an exaggerated attributional bias. CONCLUSIONS: The results suggest that depressed patients tend to blame themselves for their mistakes. Depressed patients made more stable and global attributions for negative events than for positive events, as opposed to what happens with the normal controls, making us believe that this bias will be maintained in time, and will eventually generalize to other life areas. This kind of study seems useful to develop efficient treatment programs and evaluation tools of depression. PMID- 15529210 TI - [Relationship between malingered psychometric profiles and personality styles in prisoners]. AB - INTRODUCTION: One of the most typical clinical problems is the assessment of malingering in the field of forensic and penitentiary psychiatry. OBJECTIVE: The purpose of this research was to find associations between different personality types and psychometric response styles. MATERIAL AND METHODS. SUBJECTS: participants consisted of 41 imprisoned male offenders. Instruments and data collection: the psychometric tools used were the Millon Clinical Multiaxial Inventory (MCMI-II) and the Minnesota Multiphasic Personality Inventory (MMPI-2). STATISTICS: data were analyzed with the Statistical Package for the Social Sciences (SPSS, version 10.0). A p < 0.05 significance level was proposed. RESULTS: There is significant correlation between personality types and styles of responses in the inventories. CONCLUSIONS: The greatest relationships were found between personality types shaping cluster B of DSM-IV-TR and validity indexes that measure malingering as response style. PMID- 15529211 TI - [Economic analyses of olanzapine in the treatment of schizophrenia and bipolar disorder]. AB - Schizophrenia generates important costs for society both direct, as a consequence of hospitalization and outpatient treatment, and indirect; related to loss of productivity. The atypical antipsychotics, such as olanzapine, have supposed an important advance in the treatment of schizophrenia. The greater cost of atypical antipsychotics with respect to conventional drugs has led to the conduction of pharmacoeconomic studies to determine its efficiency. This article reviews the complete pharmacoeconomic studies that compare olanzapine with haloperidol and risperidone in the treatment of schizophrenia. Cost analyses comparing olanzapine and haloperidol show that the former drug does not add increased cost to therapy, and even contributes to lessen expenses fundamentally as a result of a decrease in hospitalizations. In the economic evaluations comparing olanzapine and risperidone, the results are not conclusive, and in general, the total costs associated with both treatments were similar. In the treatment of bipolar disorder, although few studies have estimated the economic impact of olanzapine, it has been observed a reduction of hospitalization costs associated to the treatment with olanzapine. PMID- 15529212 TI - [Once weekly fluoxetine, tolerability and safety according to use patterns in the psychiatric clinical practice]. AB - INTRODUCTION: Treatment with selective serotonin reuptake inhibitors (SSRI) has become generalized in all the medical setting although its efficacy and good tolerability may be affected by long term non-compliance. Once weekly fluoxetine could improve compliance due to the comfort of its use. The objective is to evaluate the safety, tolerability and efficacy of once weekly fluoxetine (90 mg) in a sample of out-patients, following naturalistic criteria in the usual clinical practice. METHOD: A total of 46 patients (age: 41.89 +/- 1.85) who received once weekly fluoxetine for at least 3 months (mean time: 135.24 days) were analyzed retrospectively. Clinical Global Impression of Severity (CGI-S) at the onset and end of the period evaluated, presence of adverse events, drop-out index and impression of patient's satisfaction with the treatment and use pattern were used. RESULTS: The mean of the final CGI-S was 2.09, compared with the onset 3.09 (p < 0.02), indicating significant improvement in the efficacy with once weekly fluoxetine. The most frequent adverse effects were: anxiety (10.87 %), headache (8.69 %) and restlessness (8.69 %) related with once weekly fluoxetine. Compliance was very high at the onset (99 %) and at the end (96 %). Ten patients dropped-out of the treatment (27.74 %), 6 due to appearance of undesirable effects (anxiety, headache and insomnia), three due to voluntary wish and only one due to lack of efficacy. The CGI of satisfaction of clinical global efficacy on the final visit was 1.43 (satisfied-very satisfied) and the CGI of satisfaction for the treatment pattern was 1.17 (very satisfied). CONCLUSION: Change to once weekly fluoxetine generally improves satisfaction of treatment efficacy and its use pattern, although some patients return to the initial regime after adverse effects appear. PMID- 15529213 TI - [Impact of medical counseling to quit smoking during the process of dehabituation in patients with mental illness]. AB - BACKGROUND: Cigarettes consumption is the most preventable cause of morbidity and mortality in developed countries. Several intervention trials have shown that medical counseling to quit smoking promotes 5-10 % of abstinence per year, which constitutes the most efficient preventable activity. This study aims to evaluate the presence and type of medical counseling to quit smoking in patients admitted to a long-stay unit of a psychiatric hospital. METHODS: This descriptive and cross-sectional study included 80 schizophrenic smoking in-patients. The presence of medical counseling to quit smoking was evaluated by a standardized questionnaire designed for this purpose. The Fagerstrom Test was used to measure nicotine dependence, the Richmond Questionnaire to measure motivation for smoking cessation and the Stages of Change Questionnaire to know the stage of change to give up smoking. RESULTS: A total of 60.3 % of the patients had previously received medical counseling to quit smoking, 50 % of which had been performed by the psychiatrist. The motivation for patients with medical counseling was moderate (mean score +/- SD of Richmond Questionnaire 4.4 +/- 3.5). A total of 77.1 % of these patients had tried to quit smoking on several occasions. CONCLUSIONS: Medical counseling on giving up smoking can modify the motivation to quit smoking in psychiatric patients. Systematic structured counseling, especially from psychiatry, can prevent the development of nicotine addiction or modify intensity. PMID- 15529214 TI - [Validation of a Spanish version of the Diagnostic Interview for Bordelines Revised (DIB-R)]. AB - INTRODUCTION: Borderline Personality Disorder (BPD) is the most studied Axis II disorders. However, there are no Spanish versions of specific interviews. The Diagnostic Interview for Borderlines-Revised (DIB-R) is a semistructured interview used to determine the diagnosis and severity of BPD patients. The aim of this study was to validate the DIB-R for use in a Spanish-speaking sample. METHOD: The psychometric characteristics of the DIB-R Spanish version were assessed in a sample of 156 patients with the possible diagnosis of borderline personality disorder. There were 29 men and 127 women with a mean age of 27.6 years (SD: 6.5; range: 18-45). The Spanish adaptation of the Structured Clinical Interview for DSM-III-R Personality Disorders (SCID-II) was used as gold standard. RESULTS: The DIB-R showed good total internal consistency (Cronbach's alpha: 0.89) and high inter-rater reliability (within-class correlation: 0.94). Using logistic regression analyses the best cut-off was judged to be 6 or more, obtaining high sensitivity (0.81), specificity (0.94) and moderate convergent validity of the diagnosis with the SCID-II (kappa: 0.59). CONCLUSIONS: The Spanish version of the DIB-R showed psychometric characteristics similar to those in the original interview and may be useful to determine BPD presence and severity. PMID- 15529215 TI - [Study of effectiveness of craving control with topiramate in patients with substance dependence disorders]. AB - INTRODUCTION: Effectiveness and tolerability of topiramate at 3 and 6 months was assessed in patients requesting dehabituation programs. METHODS: Observational, prospective, national and multicenter study of 6 months, in patients on treatment with topiramate, who fulfilled criteria for dependence of opiates according to ICD-10 participating in therapeutic programs of dehabituation, without concomitant psychiatric illnesses and any responsible relative. Main measures of effectiveness were retention rates, alcohol consumption and other illicit drugs by urine tests (opiates, cannabis, cocaine) and treatment needs by EuropASI scale. Other parameters were HAM-D, DAS-SV and SF-36. RESULTS: Patients with consumption by urine tests decreased from 94.1 % (n = 64) at baseline to 39.6 % (n = 19) after 6 months of treatment, as was seen by means of the mean score in EuropASI scale, for all substances except methadone. No consumption was accompanied by a low rate of relapse of 33.3 % at 6 months. Twenty one patients had adverse reactions (28 %). The most frequent adverse reactions were somnolence (n = 9; 12 %), paraesthesia (n = 5; 6.7 %) and depression (n = 4; 5.3 %). CONCLUSIONS: In real clinical practice, topiramate showed a good response with a relevant decrease of percent of patients with abuse or consumption, and a satisfactory tolerability profile for the treatment of patients with dependence on heroine, cocaine, and other opiates, showing better outcomes than those obtained in previous trials. PMID- 15529216 TI - [Gender based psychopharmacology: gender influence in the pharmacological treatment of mental disorders]. AB - Gender-based pharmacology is a new research and clinical area which could be defined by the inclusion of gender-related variables in clinical, research and educational issues with respect to pharmacotherapy. Most of the studies which have studied pharmacokinetic gender differences in psychotropic drugs have found that women tend to have higher plasma concentrations. As regards differences in treatment response, most evidence points to a more rapid response to neuroleptics and lower therapeutic doses in women, and, on the other hand, a possible better response to selective serotonin reuptake inhibitors (SSRI) in premenopausal women. In addition, women appear to have a higher incidence of adverse effects to different psychotropic drugs. PMID- 15529217 TI - [New mood stabilizers and reproductive cycle]. AB - INTRODUCTION: The use of new mood stabilizers in the treatment of bipolar disorder has supposed a revolution, especially due to its more favorable profile in many aspects. Nevertheless, therapeutic decisions on treatment during pregnancy and the breastfeeding period are still being debated. Since these new anticonvulsants appeared in the decade of 1990, less naturalistic experience in its use exists during these periods than with other older anticonvulsants. METHODS: A Medline and Embase search was conducted from 1970 to 2003 to review the articles published on the use of the new mood stabilizers during pregnancy and breastfeeding, and its effects on contraception. Neurology and psychiatry text chapters and abstracts of the Annual Meeting of the American Psychiatric Association (years 2001-2003) were also reviewed. RESULTS: Although some recent articles suggest that new mood stabilizers could have a smaller risk of congenital defects, and therefore could be used with smaller risks in women in fertile age, most of articles reviewed indicate that there is not enough knowledge on the safety of the new mood stabilizers in pregnancy and breastfeeding. CONCLUSIONS: The risks and benefits of continuing with the new mood stabilizers during pregnancy and breastfeeding must be weighed carefully, and the severity of the disease and the previous answer to treatment should be taken into account. PMID- 15529218 TI - [Cycloid psychoses. A case report]. AB - Cycloid psychoses, described by Leonhard, have a sudden onset, unstable polymorphic delusion symptomatology, labile state of consciousness, lack of physical symptoms, quick remission with no residual mental abnormalities and normality between episodes. Leonhard distinguished three clinical forms: anxiety happiness psychosis, excited-inhibited confusion psychosis and hyperkinetic akinetic motility psychosis. The essential characteristics of this clinical subtypes are: polymorphism, global disturbance of psychic life, acute appearance of symptoms, total insomnia 3 days before onset of symptomatology, intra and interepisode lability, polar structure, tendency to repetition of episodes (phases) and a good long-term prognosis. We present a patient's clinical history and evolution that illustrate the characteristics of this kind of endogenous psychoses. PMID- 15529219 TI - [Leukopenia as side effect of paroxetine]. PMID- 15529220 TI - [National and international impact factor of Actas Espanolas de Psiquiatria]. AB - The aim of this paper is to present the bibliometric indicators of Actas Espanolas de Psiquiatria that were obtained from the study "Potential impact factor of the Spanish medical journals in 2001", financed by the Spanish Ministerio de Educacion, Cultura y Deporte. The citations made in Actas Espanolas de Psiquiatria and its national and international impact factor and immediacy index have been obtained by the use of a methodology similar to the one used by the Institute for Scientific Information. The national indicators only take into account the citations made in 87 Spanish journals considered as sources, while those from the foreign source journals of Science Citation Index have been added to the previously cited ones. Actas Espanolas de Psiquiatria has obtained a national impact factor of 0.315 and an international impact factor of 0.395, which places it as a leader in the Spanish psychiatric journals. PMID- 15529221 TI - [Polipharmacy in the antipsychotic prescribing in practices psychiatric out patient clinic]. AB - INTRODUCTION: Due to the lack of scientific studies in the psychopharmacological treatment of psychosis, the references advise against the use of antipsychotic polypharmacy (AP). This study investigates the situation of this matter and its relationship with the introduction of atypical antipsychotics (AA) in our daily out-patient practice. METHODS: The study group included 160 patients, with ICD-10 diagnosis of non-affective psychosis and illness evolution of at least two years, whose ages ranged from 18 to 65 years, without mental retardation, who came to the out patient clinic at least once during the first semester of 2002 and, in order to make a comparison, at least one other time 9 to 12 months before. The retrospective study was done using the clinical records. RESULTS: In both moments studied, more than 50 % of the patients (65 % and 63.4 % respectively) had AA as their only treatment. AP was 25.6 % and 26.9 % respectively, 19.4 % of all patients were always in AP. At the end of the study, patients on AP were being prescribed a higher dose of chlorpromazine equivalents per day (p < 0.001) and more anticholinergics (p < 0.001). CONCLUSIONS: Our results show that in out patient settings AA have been consolidated as the first treatment option and that AP is a very common and stable phenomenon. PMID- 15529222 TI - [Variability in psychiatric medical practice evaluated by studying short-term psychiatric rehospitalization]. AB - INTRODUCTION: Variability in medical practice can detect deficiencies in medical care quality. This study was designed to determine variations in psychiatric practice by studying short-term rehospitalization (SRH) that could result in differences in quality of psychiatric care. METHODS: Data on 894 consecutive admissions to an inpatient unit from March to December 2001 were collected. Readmissions over the next 90 days or less (SRH) were analyzed. The database was reviewed in order to extract information about several variables: age, gender, period from the first admission, number of previous admissions, interval between initial discharge and readmission, psychiatric diagnosis, aftercare provided by the mental health centers (MHC) and psychiatrist-associated variables. Differences between SRH and the other admissions were examined. RESULTS: There were significant variations in the aftercare provided by the MHC (p = 0.028). There were also variations in the psychiatrist responsible for the inpatient unit (p = 0.03), in the reference psychiatrist in a MHC (p = 0.007), but not in the emergency unit. CONCLUSIONS: This short-term rehospitalization study showed variations in quality of aftercare and in psychiatrist associated variables. Both these findings warrant further investigation that pays specific attention to staff attitudes, system barriers and facilitators of psychiatric care. PMID- 15529223 TI - [Cocaine and other psychostimulant consumption: their relationship with the childhood hyperactivity syndrome]. AB - INTRODUCTION: We want to know the attention deficit hyperactivity disorder (ADHD) among those subjects who require treatment due to cocaine and other psychostimulant drugs consumption. No previous Spanish studies have been found. METHODS: Two groups of male subjects were established out of a total of 109 subjects. Their ages ranged from 16 to 40 years. The groups were "experimental" (patients under treatment) and "control", who underwent a urine analysis for toxic agents. An interview elaborated for this and the E mode SCID-I were used. A total of 21 % of the cocaine consumers had a comorbid history of ADHD in childhood versus 3% of the control group. CONCLUSIONS: The data found overlap with the United States studies. ADHD appears as a new risk factor for the posterior development of cocaine abuse disorder, adding a cocaine addict patient subtype that deserves special diagnostic criteria as well as more complete treatment alternatives for these dual diagnosed patients. PMID- 15529224 TI - [Family transmission of positive and negative symptoms in familial and sporadic schizophrenia]. AB - INTRODUCTION: A genetic component is recognized as a cardinal feature in the etiology of schizophrenia that is presently conceived as a complex disease. However, identifying its molecular bases has become a problem, mainly due to the difficulties that the schizophrenic phenotype presents. The aim of this study is to determine whether the positive and negative symptoms are differentiated from each other according to family history in a family study using familial/sporadic strategy. METHODS: 601 subjects were studied (196 schizophrenics, 205 first degree relatives and 200 controls). A SCAN system and the diagnosis criteria of DSM IV were used. For the familial study the FIGS and PANSS scales were applied. Families were grouped into familial or sporadic groups according to family aggregation. Comparisons were made using the ANOVA. RESULTS: The groups of patients and the first degree relatives with familial schizophrenia showed significantly higher scores in the negative sub-scale than did the sporadic schizophrenia subjects. Behavior of the first degree relatives with sporadic schizophrenia was similar to the control group. Positive sub-scale scores and general psychopathology showed no differences in the patients and their relatives according to familiar aggregation. CONCLUSIONS: This study confirms previous results that negative symptoms are more frequent in schizophrenics and their first degree relatives with familial schizophrenia. Therefore, it can be said that negative symptoms are a clinical phenotype more related to the genetic etiology. PMID- 15529225 TI - [Beliefs About Voices Questionnaire (BAVQ): Spanish translation and psychometric properties]. AB - INTRODUCTION: The objective of this study was to translate into Spanish and to determine the inner reliability and consistency of the Beliefs About Voices Questionnaire (BAVQ) in a sample of Mexican patients diagnosed of schizophrenia and/or schizoaffective disorder under the DSM-IV criteria. METHOD: In order to do this, a diagnosis confirmation based on the semistructured interview format SCID was applied. The Spanish version of the BAVQ and Beck's anxiety and depression inventories were also implemented with a total of 55 persons, including males and females between 18 and 65 years old consecutively referred by the Mental Health Care Centers belonging to Mental Health Institute of Jalisco which forms a part of the governmental Health Care System in Mexico. RESULTS: The inner consistency indicators from the Spanish version ranged from 0.74 to 0.88, similar to those reported in the original version reviewed. The total score in the Malevolence and Resistance subscales were positively correlated and were statistically significant with depressive symptoms evaluated with the Beck Depression Inventory (BDI) (r = 0.29, p = 0.04; r = .35, p = 0.01, respectively), as well as the "omnipotence" subscale with the presence of anxiety symptoms according to the Beck Anxiety Inventory (BAI) (r = 0.35; p = 0.01). The factorial structure was similar to that reported for the original version of the instrument reviewed. CONCLUSIONS: The Spanish version of the BAVQ is a valid and reliable instrument to evaluate beliefs about identity, power, purpose, and consequences of obeying auditory hallucinations or "voices" in the Spanish speaking population. PMID- 15529226 TI - [Chinese Polarity Inventory: its adaptation to our setting]. AB - INTRODUCTION: The Chinese Polarity Inventory (CPI) is a self-report scale for bipolar disorder assessment, composed of two scales (mania and depression), with 20 items for each one. OBJECTIVES: Translate, adapt and validate the CPI to our context. MATERIAL AND METHODS: The questionnaire was applied to 123 patients: bipolar disorder in the manic phase (32), depressed (17), remission (30) and mixed state (3). We also established a control group with 30 patients without affective disorder. Internal consistency was assessed using Cronbach's alpha. On the other hand, validity was established using the Clinician Administered Rating Scale for Mania (CARS-M) in manic patients and the Hamilton scale in the depressed ones as a gold standard. RESULTS: This scale has good psychometrics qualities. Internal consistence index (Cronbach's alpha) for the manic scale was 0.90 and for depression scale was 0.92. The correlation coefficient between the manic scale and the CARS-M was 0.82 and between the depression scale and the Hamilton scale was 0.67. CONCLUSIONS: The CPI is an instrument of rapid and easy application that permits self-report of bipolar disorders. The best benefit for this scale is the possibility of application in the entire course of illness, for clinical and investigation activities and assessment of the treatment response. PMID- 15529227 TI - [Knowledge and perception about depression in the Spanish population]. AB - INTRODUCTION: The aim of this study is to investigate the Spanish people's perception of depression. Identification of the existing mistakes in depression perception will be the first step to correct them and to contribute to the patients' well being. MATERIAL AND METHODS: More than 1,000 structured surveys were conducted all around Spain with segmented representation for each regional community. RESULTS: Two out of three interviewed people considered depression to be a disease, in a somewhat higher degree those who had suffered from depression (67%). Forty two percent of people thought depression affects more than 25% of the population. Spanish people think that the incidence of depression will increase in the future. Their opinion of the importance of depression is high, considering it among the three most prevalent diseases, even exceeding that of coronary diseases. Most people interviewed would not accept a pharmacological treatment of at least one year of duration. The most mentioned adverse effect of antidepressants is dependency, and it is the reason why interviewed people are reluctant to maintain long term treatment. Only 50% of people treated for depression thought they had had enough family and social support. Eighteen percent of depressed people hide their diagnosis, basically for fear of stigma at work. CONCLUSIONS: Nowadays, the Spanish population has a good knowledge of depression. However, there are still some prejudices that make more difficult both the treatment and the recovery of depressed people and their social readjustment. These beliefs make up small stigmas that hinder treatment adherence, which is doubtlessly the most important factor to improve the outcome of depression. PMID- 15529228 TI - [Emergence and brain development of executive functions]. AB - Development of executive functions (EF) during childhood and adolescence is closely related to frontal lobe maturation and its connections with other cortical and subcortical structures. The main maturative processes are myelination and synaptic pruning, both of which work on the brain following a hierarchical model. Different studies agree with the fact that EF emerge at the age of 6 years. However, these studies have used complex neuropsychological tests, which require appropriate functioning of several cognitive functions. This is why differential development of different EF components cannot be observed. To do so, other studies have designed simpler tasks, which only need active maintenance of information and inhibition, both basic functions for the appropriate execution of more complex EF tasks. When these simple tasks are used, an early appearance of EF can already be noticed at the age of 12 months, and there are important advances between the third and fifth year. Consequently, the idea of the frontal lobe being "functionally silent" until adolescence seems definitively discarded. PMID- 15529229 TI - [Historical approach to reserpine discovery and its introduction in psychiatry]. AB - Reserpine, an alkaloid of the Rauwolfia serpentina plant isolated during the middle of the 20th Century, represented a highly important clinical advance in the treatment of schizophrenia whose pharmacological tools were limited to chlorpromazine that was introduced in the clinical area two years before. Both agents would come into the history as the drugs that made possible the beginning of the psychopharmacological era. In the present article, a revision is made of the complicated process leading to the isolation and synthesis of reserpine, by the Swiss pharmaceutical company Ciba (Schlittler and Muller) and how its pharmacological properties (Bein) were discovered and studied, in the animals laboratory, mainly, the "tranquillizers". The introduction of this antipsychotic in psychiatry, which was initiated in 1954 (half a century ago), and the results obtained in the first clinical studies, as well as the role played by researchers such as Kline, Delay, Noce, Hollister, Altschule, etc. are then described. In addition, and from a historical perspective, the discovery of the adverse effects of this drug, especially those of extrapyramidal nature (dyskinesia and akathisia), are studied, concluding with the reasons that produced its rapid clinical decline, among which the genesis of depressive pictures (phenomenon presently questioned) may be emphasized. Finally, the conclusion reached was that, although the clinical relevance of reserpine was not as evident and long lasting as chlorpromazine, its initial contribution to the treatment of schizophrenic patients was of maximum importance. PMID- 15529231 TI - [eFirst -- Citeable online-publication ahead of print]. PMID- 15529230 TI - [A case of chronic mania]. AB - In this paper, we present a case of chronic mania and we revise its diagnosis and treatment. A case of a 23 year old patient with chronic mania as well as his response to clozapine treatment is described. PMID- 15529232 TI - [New uncertainties in legal psychiatry]. PMID- 15529233 TI - [The term insight in forensic practice]. AB - The paper comments the German legal and forensic-psychiatric positions regarding the complex and yet unsolved question of insight in the context of criminal responsibility. Possible restrictions of insight are discussed. Finally a solution is suggested, which connects the assessment of insight with the ability to declare a free will: Both terms deal with the predecisional cognitive and motivational foundation of acts. Insight is closely associated with the competence to constitute a free will. Symptoms and disorders which tend to disturb this ability can be assessed by psychopathological methods. Possible restrictions concern mental retardation, dementia, organic personality disorders, mental disorders with delusional ideation and residual states of schizophrenia. PMID- 15529234 TI - [Pathology of the thalamus and schizophrenia--an overview]. AB - Since the beginning of the 20 (th) century, the thalamus was regarded as a brain region which may be involved in the pathogenesis of schizophrenia. Distinct thalamic nuclei were morphologically analyzed with qualitative methods with an emphasis on the mediodorsal nucleus. However, the reported results were inconsistent. After the introduction of quantitative neuroanatomical methods, it became obvious that the volume and cell reductions are not only present in the association nuclei, but also in the limbic (N. anteroventralis) and motor thalamic nuclei (N. ventrolateralis posterior). The involvement of distinct thalamic nuclei is supported by structural MRI studies which have shown a moderate but significant volume reduction of the whole thalamus in this disease. The majority of fMRI and PET studies reported a reduction of the metabolic activity or blood flow in the thalamus in patients with schizophrenia. The similarity between the structural changes in animal models of thalamic plasticity and the structural thalamic alterations in schizophrenia suggest an involvement of neuroplasticity mechanisms in the pathogenesis of these alterations. Post mortem studies and In-vivo receptor studies suggest altered glutamatergic, dopaminergic and membrane-associated mechanisms within thalamic pathology in schizophrenia. On the psychopathological level, there is a similarity between some of the psychic manifestations of thalamic lesions and symptoms of the schizophrenic disease. There are also reports on volume reduction of the whole thalamus in first-episode neuroleptic-free patients. It appears unlikely that the neuroleptic medication plays an etiological important role, since no significant correlations were found between the volume and cell reductions and the neuroleptic treatment period. The reviewed data suggest that distinct thalamic nuclei and therefore distinct neuronal circuits are involved in the pathogenesis of schizophrenia. PMID- 15529235 TI - [Pathways to care: help-seeking behavior in first-episode psychosis]. AB - Several studies on first episode schizophrenia suggest that a longer duration of untreated psychosis (DUP) results in poorer clinical outcome. The same is expected for the duration of untreated illness (DUI). It is therefore important to expose people at risk of schizophrenia to adequate treatment early on. The improvement of pathways to adequate treatment within the health care system might well be helpful. Therefore, an analysis of the pathways to care is necessary. Thus, in this present study 80 in-patients with first episode psychosis were investigated using the semi-structured interview IRAOS ("Interview for the Retrospective Assessment of the Onset of Schizophrenia") and data about the pathways to care before psychiatric admission were collected. The results indicate that patients contact on average three carers. Contact to a psychiatrist or a psychotherapist was only made two and a half years after onset of illness; general practitioners were only contacted after more than five years. However, there was a significant time lapse between first contact to psychiatrist/ psychotherapist and psychiatric admission. Only 31 % of patients sought help in the prodromal phase of the illness. Two strategies for a public campaign can be derived from these results: firstly, a public awareness campaign has to be implemented to shorten the interval from onset of illness to first help-seeking behaviour and secondly, professionals need to have more knowledge and better awareness of prodromal signs in order to reduce the time between diagnosis and adequate treatment. PMID- 15529236 TI - [Neuropsychological testing in vocational rehabilitation for subjects with schizophrenia]. AB - Until now it is difficult to predict the success of vocational rehabilitation programs for subjects with severe mental illnesses. The growing knowledge about the neuropsychological mechanisms of psychiatric disorders has not been fully integrated into psychiatric rehabilitation research. We reviewed the literature how to use neuropsychological parameters for predicting outcome of vocational rehabilitation programs in subjects with schizophrenia and schizoaffective disorders. Twenty studies were reviewed, which evaluated neuropsychological variables to predict dimensions of rehabilitation success. As a result of this review, we suggest implementing neurocognitive tests, which assess executive functions, working memory, and attention deficits in routine rehabilitation to better plan and coordinate rehabilitation, and to predict rehabilitation outcome. PMID- 15529237 TI - [Sinus vein thrombosis]. PMID- 15529239 TI - [Performance and costs of outpatient clinics of university hospitals for medical care, teaching, research activities]. AB - BACKGROUND: Outpatient clinics of university hospitals (Hochschulambulanzen) play a significant role in the German health care system. Universities have in contrast to other hospitals the right to implement an outpatient clinic, but the health care services they can render are restricted to clinical research and teaching activities. The university outpatient clinic study evaluates the intensity of medical care, teaching, research activities, and the related costs. METHOD AND DATABASE: 6 university hospitals with 51 outpatient departments in Germany were included. The prospective documentation of consultations was restricted to 800 visits per department. A total of 26,312 consultations with approximately 40,000 diagnoses and 150,000 services were documented. Furthermore, data concerning costs, teaching activities and research facilities were documented. RESULTS: Clinical treatment without any correlation to research or teaching activities amounted to about 81 % of the working time in the outpatient department (research 11 %; teaching 8 %). The primary task of the university outpatient clinics takes up less than 20 % of the working time. The physicians documented that the disease of every fourth visit was in accordance with their main field of research. 6.9 % of the visits were asked to take part in clinical trials, of these 1.25 % were included for the first time, 3.7 % were already included. 6.5 % of the visits were addressed to participate in specific teaching activities. The average total costs per case added up to 149 Euro. No outpatient clinic could cover the total per case costs with the lump sum payments. On the average 31 % of these costs were covered by lump sum payments (without cases concerning research and teaching). CONCLUSION: Treatment in outpatient departments of university clinics is far beyond research and teaching activities required by law. However, the ability of outpatient departments of universities to provide excellent outpatient services should have a more dominant role in the health care system. Therefore access to care should be deregulated for the patients and reimbursement schemes should be adjusted to adjust for the present losses. PMID- 15529240 TI - [Acute ischemia after intraarterial drug injection]. AB - HISTORY AND CLINICAL FINDINGS: A 36-year-old patient presented with a severe hand ischemia after intraarterial injection of four dissolved tablets of Methylphenidate each 10 mg (Ritalin into the right radial artery. INVESTIGATIONS: Non-invasive vascular diagnostic demonstrated a normal perfusion of the radial and ulnar artery and a severe ischemia of the small vessels of the right hand. TREATMENT AND COURSE: Under suspicion of a combined vaso-spastic and thromboembolic arterial occlusion we started an intraarterial lysis therapy followed by anticoagulation with heparin and infusions of prostaglandines. Despite this therapy necroses of three fingers developed; partial amputation was followed by a good wound healing. CONCLUSIONS: Ischemia of an extremity after intraarterial injection of drugs is a vascular emergency. PMID- 15529241 TI - [Autoimmune cholangitis with vasculitic ulcers, rheumatoid arthritis and IgA glomerulonephritis]. AB - HISTORY AND ADMISSION FINDINGS: A 51-year-old woman was admitted because of relapsing episodes of fever and leg ulcers for 14 years. In addition, she had polyserositis, polyarthralgias and polyarthritides, renal failure with proteinuria and elevation of gamma-GT and alkaline phosphatase. The patient was in a reduced general condition and cachectic nutritional state. She had slight scleral icterus, the liver being palpable 5 cm under the costal margin, edema of the lower limbs and two ulcers at the right foot. INVESTIGATIONS: Laboratory examinations revealed leukocytosis, anemia, elevation of cholestasis and inflammation parameters as well as renal failure. During a 24 hour collection period, a significant proteinuria was demonstrated. Immunoserologically, an ANA titer of 1:100 and a positive rheumatoid factor were found, ANCAs and AMAs were negative. On ultrasound, both kidneys exhibited a blurred pelvic parenchymal border. Thyroid ultrasound demonstrated parenchymal changes consistent with Hashimoto's disease. Ultrasound of the wrist revealed extensive arthritis with tendovaginitis. A renal biopsy revealed mesangioproliferative glomerulonephritis. DIAGNOSIS, THERAPY AND CLINICAL COURSE: Due to serologically persistent cholestasis, a liver biopsy was performed which, together with negative AMAs, revealed the diagnosis of an autoimmune cholangitis (AIC; AMA-negative primary biliary cirrhosis (PBC)). In addition, the patient presented with rheumatoid arthritis, polyserositis, IgA glomerulonephritis, vasculitic leg ulcers and Hashimoto's thyreoiditis which were interpreted as extra-hepatic manifestations of the AIC. After initiation of high dosage corticosteroid therapy, rapid healing of the leg ulcers occurred. Therapy of the AIC consisted in ursodeoxycholic acid. CONCLUSION: The multitude of associated immunological phenomena in this patient resulted in a delay of the diagnosis. A AIC/PBC, however, should, always be considered in case of cholestasis. PMID- 15529244 TI - [Cardiogenic shock -- case report, diagnostics, therapy]. PMID- 15529248 TI - [Readmission regulation based on a complication (KFPV 2004) in the DRG appraisal practice]. PMID- 15529247 TI - [Current strategies in the treatment of advanced stage mantle cell lymphoma]. AB - Advanced stage mantle cell lymphoma (MCL) with a median survival of only three years and virtually no long-term survivors represents the lymphoma subtype with the poorest prognosis and remains incurable with conventional chemotherapy. Recently two randomized trials of the German Low Grade Lymphoma Study Group (GLSG) demonstrated the superiority of a combined immunochemotherapy with the anti-CD20 antibody rituximab in first-line therapy (R-CHOP) as well as in relapsed disease (R-FCM). In addition, in a trial of the European MCL Network, intensified-consolidation with high-dose radiochemotherapy followed by autologous stem cell transplantation significantly improved the progression-free survival in patients up to 65 years of age. However, the vast majority of patients with MCL will eventually relapse. Thus, new strategies such as allogenic transplantation after dose-reduced conditioning or novel molecular targeting agents (e. g. proteasome inhibitors or radiolabeled antibodies) are urgently warranted to further improve the long-term outcome of MCL. PMID- 15529249 TI - [Readmission regulation in the DRG system: a complete consideration]. PMID- 15529250 TI - Ebola virus ecology. PMID- 15529251 TI - A serological survey of Ebola virus infection in central African nonhuman primates. AB - We used an ELISA to determine the prevalence of IgG antibodies specific for the Zaire subtype of Ebola virus in 790 nonhuman primates, belonging to 20 species, studied between 1985 and 2000 in Cameroon, Gabon, and the Republic of Congo. The seroprevalence rate of Ebola antibody in wild-born chimpanzees was 12.9%, indicating that (1) Ebola virus circulates in the forests of a large region of central Africa, including countries such as Cameroon, where no human cases of Ebola infections have been reported; (2) Ebola virus was present in the area before recent outbreaks in humans; (3) chimpanzees are continuously in contact with the virus; and (4) nonlethal Ebola infection can occur in chimpanzees. These results, together with the unexpected detection of Ebola-specific IgG in other species (5 drills, 1 baboon, 1 mandrill, and 1 Cercopithecus), may help to narrow the search for the reservoir of Ebola virus. They also suggest that future Ebola outbreaks may occur anywhere in the central African forest region. PMID- 15529252 TI - Simian parvovirus infection: a potential zoonosis. AB - INTRODUCTION: Simian parvovirus (SPV) causes severe anemia in immunocompromised macaques. The closely related erythrovirus, parvovirus B19, causes anemia in susceptible humans and can be grown in human bone marrow mononuclear cells in vitro. We hypothesized that SPV may infect humans and replicate in human bone marrow mononuclear cells. METHODS: Serum samples from handlers of an SPV seropositive macaque colony were tested by Western blot for evidence of antibodies to SPV. SPV capsid protein was expressed in insect cells, and SPV was cultured in human and macaque bone marrow mononuclear cells. RESULTS: Fifty-one percent of exposed handlers (n=65) were found to be SPV seropositive, compared with 35% of nonexposed individuals (n=124). In 17% of exposed handlers, compared with 6% of nonexposed individuals, antibodies were directed to SPV but not to B19. SPV capsid proteins, like those of B19, self-assembled to form parvovirus like particles, and these capsids, like B19 capsids, bound to globoside, suggesting that globoside is also the receptor for SPV. We demonstrated that SPV could replicate in vitro in both human and macaque bone marrow mononuclear cells and that it was cytotoxic to erythroid progenitor cells. CONCLUSIONS: Our data suggest that SPV may infect human bone marrow mononuclear cells in vitro and in vivo and should be considered a potential zoonosis. PMID- 15529253 TI - Antibody maturation and viremia after primary cytomegalovirus infection, in immunocompetent patients and kidney-transplant patients. AB - To investigate antibody maturation and serum levels of cytomegalovirus (CMV) DNA after primary CMV infection, we studied 51 immunocompetent and 27 kidney transplant patients. Compared with the immunocompetent patients, the transplant patients had significantly more-prolonged and -variable antibody maturation, clearly longer durations of viremia, and higher levels of CMV DNA; however, antibody maturation continued for >1 year even in immunocompetent patients. Long term ganciclovir prophylaxis in the transplant patients was associated with either delayed immunoglobulin-G seroconversion, inhibition of antibody maturation (n=2), or immunoglobulin-class switching (n=1). In conclusion, antibody maturation continues in immunocompetent patients for a period longer than previously had been thought and is significantly delayed or even inhibited in kidney-transplant patients. PMID- 15529255 TI - Reduced numbers and impaired ability of myeloid and plasmacytoid dendritic cells to polarize T helper cells in chronic hepatitis C virus infection. AB - Hepatitis C virus (HCV) infection induces a wide range of chronic liver injuries. The mechanism by which HCV evades the immune surveillance system remains obscure. Blood dendritic cells (DCs) consist of myeloid and plasmacytoid subsets that play distinct roles in the regulation of antivirus immune responses; however, their roles in the pathogenesis of HCV infection are yet to be determined. We compared the numbers and functions of myeloid and plasmacytoid DCs between 43 patients with chronic hepatitis and 26 age-matched healthy volunteers. Absolute numbers of myeloid DCs, plasmacytoid DCs, and DC progenitors in the periphery were significantly lower in patients with chronic hepatitis than in healthy volunteers. Myeloid and plasmacytoid DCs from the patients had impaired abilities to stimulate allogeneic CD4 T cells and to produce interleukin (IL)-12 p70 and interferon- alpha , compared with those from healthy volunteers. After exposure to naive CD4 T cells, myeloid DCs from the patients were less able to drive the T helper type 1 response, whereas myeloid and plasmacytoid DCs from the patients primed more IL-10-producing cells than did those from healthy volunteers. In conclusion, in chronic HCV infection, both types of blood DCs are reduced and have an impaired ability to polarize T helper cells. PMID- 15529254 TI - Use of the P gene to genotype human metapneumovirus identifies 4 viral subtypes. AB - This study, conducted during 2001-2003, undertook the screening of patients with acute infectious respiratory-tract disease. A random selection of positive specimens was used for sequencing studies of the human metapneumovirus (hMPV) nucleoprotein gene and the phosphoprotein (P) gene. Australian and international sequences were compared, and a global classification scheme was developed. The hMPV P gene was an ideal molecular target for the detection and genotyping of hMPV. The region contained conserved sequences for reverse-transcriptase polymerase chain-reaction primers and adequate variability to permit the accurate genotyping of the virus into 2 main lineages and 4 sublineages. Establishing viral identity is essential for the linking of genotype and disease severity. PMID- 15529256 TI - Parenteral influenza vaccination influences mucosal and systemic T cell-mediated immunity in healthy adults. AB - We sought to determine whether palatine tonsils (PTs) harbor naturally acquired influenza-specific T cell immunity and whether routine parenteral immunization with influenza vaccine influences mucosal and systemic T cell reactivity. We demonstrate that tonsillar and peripheral blood mononuclear cells (PBMCs) proliferate strongly to influenza antigens, suggesting that naturally acquired immunity exists within both the mucosal and systemic compartments. Influenza vaccination induced significantly stronger T cell responses in both PTs and blood, in addition to increasing titers of anti-influenza antibodies in serum and saliva. More-rapid proliferative responses of PTs after vaccination were associated with a shift from a response involving both CD45RA+ and CD45RO+ T cells to an entirely CD45RO+-dependent response. Interestingly, the ratio of interferon- gamma to interleukin-5 was dramatically higher in cultures of PT T cells responding to influenza than in PBMCs. Our data indicate that parenteral influenza vaccination influences both mucosal and systemic naturally acquired T cell immunity. PMID- 15529257 TI - Anti-G protein antibody responses to respiratory syncytial virus infection or vaccination are associated with inhibition of G protein CX3C-CX3CR1 binding and leukocyte chemotaxis. AB - Respiratory syncytial virus (RSV) is an important cause of severe lower respiratory tract illness in infants and the elderly. Presently, no safe and efficacious RSV vaccine exists; however, advances in our understanding of immunity and the pathogenesis of disease associated with RSV infection may lead to new vaccine strategies. RSV G protein contains a CX3C chemokine motif that interacts with the CX3CR1 chemokine receptor and modifies the activities of fractalkine. In the present study, we show that anti-RSV G protein antibody responses after recent RSV infection or vaccination are associated with inhibition of RSV G protein CX3C-CX3CR1 interaction and RSV G protein-mediated leukocyte chemotaxis. PMID- 15529258 TI - Variable resistance to palivizumab in cotton rats by respiratory syncytial virus mutants. AB - BACKGROUND: Palivizumab (PZ) is the only monoclonal antibody in use against a human infectious disease. PZ is given as prophylaxis against infection with respiratory syncytial virus (RSV). An RSV escape mutant, MP4, has been shown to resist PZ prophylaxis in cotton rats. METHODS: To further define the potential of RSV to resist prophylaxis, additional PZ-resistant viruses were selected in cell culture and were tested for susceptibility to PZ in cotton rats. RESULTS: Mutant MS412 had an A-->C mutation at nucleotide position 827 in the F gene, resulting in an amino acid change from Lys to Gln at position 272. Mutant F212 had an A-->T mutation at position 816, leading to an amino acid change from Asn to Ile at position 268. In vitro, F212 had impaired growth kinetics. In cell culture, F212 was partially and MS412 was completely resistant to PZ neutralization. A single prophylactic dose of 15 mg/kg PZ protected cotton rats from infection with F212 but not with MS412. CONCLUSION: Both in vitro and in vivo, individual RSV PZ escape mutants varied in their susceptibility to PZ. Mutations associated with resistance to PZ did not always result in failure of PZ prophylaxis. PMID- 15529259 TI - Time to virological failure of 3 classes of antiretrovirals after initiation of highly active antiretroviral therapy: results from the EuroSIDA study group. AB - OBJECTIVE: The purpose of the present study was to determine the prevalence and incidence of virological triple drug-class failure (TCF) and to summarize the clinical outcome for patients who started receiving highly active antiretroviral therapy (HAART). METHODS: The present study is an observational longitudinal study of 3496 treatment-experienced (TE) and treatment-naive (TN) patients monitored from the time they started receiving HAART (baseline) until TCF occurred (as determined on the basis of viral loads), until AIDS was newly diagnosed, or until death. RESULTS: Four hundred forty-five patients (12.7%) had TCF; 370 (16.6%) of 2230 patients were TE, and 75 (5.9%) of 1266 patients were TN. At 6 years after starting HAART, 21.4% of TE and 11.2% of TN patients had TCF (P<.0001). The prevalence of TCF at or after 2002 was 15.5% in TE patients and 4.8% in TN patients. TN patients had a 32% annual increase in the incidence of TCF (95% confidence interval [CI], 14%-54%; P<.0001); at 5 years after starting HAART, the rate was comparable for TE and TN patients (3.3 and 3.4 cases/100 person-years of follow-up [PYFU], respectively). The incidence of new cases of AIDS or death was 2.7 cases/100 PYFU in patients who did not experience TCF and 5.0 cases/100 PYFU in patients who did experience TCF, an estimated 36% increase with each category of TCF (95% CI, 19%-56%; P<.0001). CONCLUSION: The prevalence of TCF was low after patients started receiving HAART, particularly among TN patients. Despite the influx of patients who had started receiving HAART more recently, the prevalence of TCF increased over calendar time. Patients with TCF had a higher incidence of newly diagnosed AIDS or death. Treatment of patients with TCF deserves further investigation. PMID- 15529260 TI - Capravirine, a nonnucleoside reverse-transcriptase inhibitor in patients infected with HIV-1: a phase 1 study. AB - Capravirine is a nonnucleoside reverse-transcriptase inhibitor (NNRTI) with a unique resistance profile. Although single mutations allow resistance to established NNRTIs, human immunodeficiency virus (HIV)-1 must undergo multiple mutations to achieve resistance to capravirine. In the present phase 1 study, capravirine was administered orally for up to 28 days to 55 HIV-1-infected individuals with CD4+ T lymphocyte counts of 50-500 cells/microL. The most frequent adverse events were diarrhea (5%) and nausea (4%), with no drug-related rashes observed. The day 15 median (mean) HIV-1 load decreased by 1.34 (1.45) log(10) copies/mL in the patients receiving 25 mg/kg/day. Capravirine demonstrated potent antiviral activity, even in antiretroviral-experienced patients. PMID- 15529261 TI - Demographic factors that influence the neutralizing antibody response in recipients of recombinant HIV-1 gp120 vaccines. AB - We compared neutralizing antibody responses in human immunodeficiency virus (HIV) type 1 gp120 vaccine recipients by age, sex, and race. Four phase 1 or 2 trials involving 505 vaccinated subjects were analyzed. Age and sex had no detectable effect on neutralizing antibody responses. However, race influenced the response to one vaccine, MN gp120, in alum. Four inoculations with this vaccine generated higher serum titers of neutralizing antibodies in African Americans than in whites. Despite potent neutralization of T cell line-adapted HIV-1, serum from these African Americans failed to neutralize primary HIV-1 isolates. Neutralizing antibody responses did not differ between races when SF2 gp120 in MF-59 was administered either alone or with recombinant canarypox vCP205; they also did not differ when vCP1452 was administered either alone or with AIDSVAX B/B in alum. These data indicate that race may affect the neutralizing antibody response to some gp120 immunogens. To fully evaluate immunogenicity, clinical trials of candidate vaccines should enroll diverse populations of subjects. PMID- 15529263 TI - Cellular viral rebound after cessation of potent antiretroviral therapy predicted by levels of multiply spliced HIV-1 RNA encoding nef. AB - To characterize newly arising replication of human immunodeficiency virus (HIV) type 1 in vivo at the cellular level, distinct viral RNA species in peripheral blood mononuclear cells (PBMCs) from HIV-1-infected patients were monitored during 2 weeks of structured treatment interruption (STI). HIV-1 RNA encoding tat/rev and PBMC-associated virions were almost completely depleted during antiretroviral therapy and emerged simultaneously after 2 weeks of STI, thus specifically reflecting productive viral infection at the cellular level. The magnitude of these correlates of reappearing cellular viral replication was predicted by during-therapy levels of nef transcripts in PBMCs. Significant rebound of plasma viremia, representing the progeny of a broader range of anatomical compartments, preceded and predicted productive infection in PBMCs. Thus, cellular viral rebound in PBMCs likely was primed before STI by the expression of nef in HIV-1-infected PBMCs that lacked virion production and was subsequently triggered by the plasma viremia that preceded the recurrence of productively infected PBMCs. PMID- 15529262 TI - Women with cervicovaginal antibody-dependent cell-mediated cytotoxicity have lower genital HIV-1 RNA loads. AB - Antibodies that mediate human immunodeficiency virus (HIV)-specific antibody dependent cell-mediated cytotoxicity (ADCC) are present in the cervical fluid of many HIV-positive women; however, the role that these antibodies play in host defense against HIV is not known. To understand the contribution of ADCC in cervical secretions as a protective mechanism against HIV, we evaluated ADCC titers in paired serum and cervical-lavage (CVL) samples from >300 HIV-1-positive women who participated in the multicenter Division of AIDS Treatment Research Initiative Study 009. The present study demonstrates that women with CVL ADCC activity had lower genital viral loads than did women with serum ADCC activity only. Women with CVL ADCC activity were likely to have HIV-1 gp120-specific immunoglobulin (Ig) G, but not IgA, in their cervical fluid. This finding suggests that specific IgG in cervical fluid can mediate ADCC activity that inversely correlates with genital viral load. PMID- 15529264 TI - A model system of oral HIV exposure, using human palatine tonsil, reveals extensive binding of HIV infectivity, with limited progression to primary infection. AB - Oral exposure to human immunodeficiency virus (HIV) type 1 results in systemic infection, but many details surrounding virus transmission remain unresolved. We developed a mucosal model, using human palatine tonsil with intact external epithelium, to study events after oral exposure to HIV. When applied to the external epithelium, semen from an HIV-seropositive patient and cell-free virus both established HIV infection in individual tonsillar cells. However, clusters of infected tonsillar cells were detected where the epithelial surface was damaged. Investigation of the initial events in HIV transmission revealed extensive and stable binding of HIV virions and seminal cells to tonsil epithelium. In experiments modeling physiologically relevant events, the addition of seminal plasma resulted in enhanced virion binding to epithelial cells. These results indicate that, although extensive binding of HIV virions and seminal cells can be demonstrated at an exposed mucosal surface, there is only limited progression from binding to primary infection. PMID- 15529265 TI - Antiretrovirals as antimalarial agents. AB - Recent studies have indicated that antiretroviral protease inhibitors may affect outcome in malarial disease. We have investigated the antimalarial activities of 6 commonly used antiretroviral agents. Our data indicate that, in addition to the previously published effects on cytoadherence and phagocytosis, the human immunodeficiency virus (HIV)-1 protease inhibitors saquinavir, ritonavir, and indinavir directly inhibit the growth of Plasmodium falciparum in vitro at clinically relevant concentrations. These findings are particularly important in light of both the high rate of malaria and HIV-1 coinfection in sub-Saharan Africa and the effort to employ highly active antiretroviral therapy in these regions. PMID- 15529266 TI - Variable adhesion abilities and overlapping antigenic properties in placental Plasmodium falciparum isolates. AB - BACKGROUND: Pregnancy-associated malaria is characterized by selection and multiplication, in the placenta, of a distinct population of Plasmodium falciparum expressing particular variant surface antigens (VSAs) that adhere to chondroitin sulfate A (CSA). METHODS: The adhesion of 40 freshly collected placental parasite isolates to bovine CSA and human placental low-sulfated chondroitin proteoglycans (CSPGs) was investigated. Plasma samples from 30 pregnant women were used to test, by flow cytometry, their recognition of and their adhesion-inhibition capacity toward 6 of these isolates. RESULTS: Adhesion to CSA and CSPGs varied between isolates but was strongly correlated between receptors (P<.001). Adhesion of isolates to receptors strongly and negatively correlated with low birth weight (LBW) of the neonate (odds ratio [95% confidence interval], 5.2 [1.1-25.1]). In plasma samples from pregnant women, the level of specific immunoglobulin G against each placental isolate (anti-VSA(PAP)) strongly correlated with the level of anti-VSA(PAP) antibodies against all other isolates (P<.05) and increased with parity in all isolates (P<.01). Conversely, adhesion inhibitory antibodies did not correlate with isolates or with the level of anti VSA(PAP) antibodies. CONCLUSION: The level of adhesion of placental parasites to chondroitin sulfate receptors is an important risk factor for LBW. Parasite heterogeneity suggests that they are composed of mixed adhesion phenotypes capable of inducing immune responses to a range of different and overlapping targets. PMID- 15529267 TI - Antibodies directed against O-acetylated sialoglycoconjugates accelerate complement activation in Leishmania donovani promastigotes. AB - BACKGROUND: An enhanced presence of 9-O-acetylated sialoglycoconjugates (9-O AcSGs) triggers the alternate pathway (AP) in Indian visceral leishmaniasis (VL). Antibodies directed against these epitopes are present in high titers. The biological relevance of these antibodies, with regard to activation of the classical pathway (CP), was investigated. METHODS: Complement activators were affinity purified, complement activation via the CP, AP, and lectin-mediated complement pathway was measured by use of an anti-C3 radio-binding assay, and the number of C3 molecules was quantitated by Scatchard analysis. Cell death induced via the complement pathways was measured by use of MTT (tetrazolium salt 3- [4, 5 dimethylthiazol-2-yl] -2, 5-diphenyltetrazolium bromide) assay, and uptake of propidium iodide (PI) was measured by flow cytometry. RESULTS: Anti-O-AcSGs from both healthy donors and patients with VL elicited C3 deposition as early as 3 min, which triggered parasite lysis, as demonstrated by use of MTT assay and corroborated by the high rate of uptake of PI. Analysis of complement activation by mannan-binding lectin and C-reactive protein demonstrated their negligible contribution during the 3-min time frame. CONCLUSIONS: Anti-O-AcSGs were identified as an important source of CP activation under normal physiological conditions, suggesting that they play a role in conferring host protection against parasite infection. PMID- 15529268 TI - Increased ratio of tumor necrosis factor-alpha to interleukin-10 production is associated with Schistosoma haematobium-induced urinary-tract morbidity. AB - Bladder and kidney disease, which affect approximately 25%-30% of subjects infected with Schistosoma haematobium, are mediated by T cell-dependent granulomatous responses to schistosome eggs. To determine why only some infected subjects develop disease, we examined the hypothesis that infected Kenyan subjects with ultrasound-detected urinary-tract morbidity (n=49) had dysregulated cytokine production leading to enhanced granulomatous responses, compared with subjects of similar age and intensity of infection without morbidity (n=100). Peripheral blood mononuclear cells from subjects with morbidity produced 8-fold greater levels of egg antigen-driven tumor necrosis factor (TNF)-alpha and had a 99-fold greater mean TNF-alpha:interleukin (IL)-10 ratio, compared with subjects without disease. No differences in cytokine response to non-egg-derived schistosome antigens were observed between groups. Subjects with morbidity had increased TNF-alpha production in response to endotoxin, suggesting an innate hyperresponsiveness. These results indicate that increased TNF-alpha production, relative to that of IL-10, is associated with developing bladder-wall morbidity with S. haematobium infection. PMID- 15529269 TI - Impact of a conjugate vaccine on community-wide carriage of nonsusceptible Streptococcus pneumoniae in Alaska. AB - BACKGROUND: Streptococcus pneumoniae is a leading cause of invasive bacterial disease and pneumonia among children. Antimicrobial resistance among pneumococci has increased in recent years and complicates treatment. The introduction of heptavalent pneumococcal conjugate vaccine (PCV7) could reduce acquisition of antimicrobial-resistant pneumococci. METHODS: We obtained 1350 nasopharyngeal swabs for culture from 1275 children aged 3-59 months presenting at 3 clinics in Anchorage, Alaska, during the winters of 2000, 2001, and 2002, as PCV7 was being introduced into the routine immunization schedule. We recorded the frequency of use of antibiotics as well as the dates of doses of PCV7 for enrolled children. We used multivariate logistic regression modeling to identify independent risk factors for overall carriage of pneumococci and carriage of PCV7-type pneumococci, cotrimoxazole-nonsusceptible (COT-NS) pneumococci, or penicillin nonsusceptible (PCN-NS) pneumococci. RESULTS: The proportion of children who were up-to-date for age, with respect to PCV7 vaccination, increased from 0% in 2000 to 55% in 2002. Carriage of PCV7-type pneumococci decreased by 43% (P<.0001). Risk of carriage of PCV7-type pneumococci was lower in 2002 than in 2000, independent of vaccination status, suggesting an indirect effect of vaccination. Carriage of COT-NS, but not PCN-NS, pneumococci also decreased (38%; P=.02), not only among vaccinated children but also among unvaccinated children without recent use of antibiotics. CONCLUSIONS: Introduction of PCV7 into the routine infant immunization schedule in a community with a high prevalence of antimicrobial-resistant pneumococci appears to reduce transmission of PCV7 vaccine serotypes and COT-NS pneumococci but has no impact on overall carriage of pneumococci or carriage of PCN-NS pneumococci. PMID- 15529270 TI - Dual phases of apoptosis in pneumococcal meningitis. AB - Significant injury during bacterial meningitis arises from mechanisms of neuronal apoptosis, particularly in the hippocampus. Apoptosis can involve both the caspase-dependent and the caspase-independent pathway, and, although both pathways have been implicated in pneumococcus-induced neuronal cell death, their relative contributions in vivo are unclear. We used mice deficient in the activation of caspase-3, ATM, and p53 to examine the role that caspase-dependent apoptosis plays in neuronal death in the context of pneumococcal meningitis. The overall symptomatology of acute infection was similar in all mice tested, indicating that late sequelae are the clinical manifestations of neuronal death. Two phases of apoptosis were discernible: neuronal injury at 18 h after infection was independent of the caspase-3 pathway, and neuronal cell death at 24 h after infection was attenuated in the absence of the caspase-3 pathway. We conclude that treatments to increase the survival rate of neurons in patients with meningitis will need to take into account at least these 2 mechanisms of damage. PMID- 15529271 TI - Quantification of human immunodeficiency virus type 1 by reverse transcriptase coupled polymerase chain reaction. PMID- 15529272 TI - HIV-1 genotypic drug-resistance interpretation algorithms need to include hypersusceptibility-associated mutations. PMID- 15529273 TI - Mutations associated with hypersusceptibility...and with good adherence. PMID- 15529276 TI - Outbreaks of genital herpes: effects on plasma HIV type 1 viral loads in individuals receiving highly active antiretroviral therapy. PMID- 15529280 TI - Getting it, together: how the nurse patient relationship influences treatment compliance for patients with schizophrenia. AB - The importance of the nurse/patient relationship is a central concept for psychiatric nursing. The present research provides a model of the nursing process identifying aspects of the nurse/patient relationship that influence treatment compliance. Five expert nurses and 15 patients participated in the study. Data collection included participant observation, field notes, memos, demographic questionnaires, interviews with the nurses post observation, and a secondary literature review. The grounded theory investigation identified a substantive theory, Getting It, Together, as the basic social process. The core category of the theory is knowing, and subcategories are socializing, normalizing, and celebrating. PMID- 15529278 TI - Mapping of the major psoriasis-susceptibility locus (PSORS1) in a 70-Kb interval around the corneodesmosin gene (CDSN). AB - Numerous putative susceptibility loci have been described for psoriasis. Among the loci confirmed in the literature, PSORS1 (the major histocompatibility complex at 6p21.3) has the strongest effect. Recent studies have highlighted a 200-kb candidate region. However, this region has not been well delimited, mainly because of the strong linkage equilibrium among the associated alleles. To finely map PSORS1, we set up a study using 17 polymorphic markers in a 525-kb interval around the human leucocyte antigen C locus (HLA-C). The results uncovered five loci with alleles strongly associated with psoriasis (Sidak-corrected P [P(c)] values from 1.8 x 10(-7) to .003), all structured in a psoriasis-susceptibility haplotype (PSH). Subsequent analysis of extended haplotypes showed that the PSH was not only present on the traditional psoriasis-susceptibility extended haplotypes (HLA-Cw6-B57, HLA-Cw6-B37, and HLA-Cw6-B13) but also on a haplotype of Sardinian origin (HLA-Cw7-B58) found to be associated with psoriasis (Pc=.0009) because of an ancestral recombination with one of the susceptibility haplotypes carrying the HLA-Cw6 allele. Comparisons of the regions identical by descent among associated and nonassociated haplotypes highlighted a minimum region of 70 kb not recombinant with PSORS1, around the corneodesmosin (CDSN) gene. PMID- 15529281 TI - Using mindfulness-based therapeutic interventions in psychiatric nursing practice -part I: Description and empirical support for mindfulness-based interventions. AB - Finding effective nursing interventions for the treatment of mental illness is a major concern for advanced practice psychiatric nurses (APPN). Increasingly, innovative psychotherapeutic treatment modalities are being used for the treatment of individuals with serious mental illness. One such innovative approach, mindfulness-based therapeutic interventions, has been shown to relieve distress for individuals with medical and psychiatric illnesses. In part one of this two-part article, the investigators describe principle concepts of mindfulness, review current research in the usefulness of mindfulness practice for treatment of psychiatric illnesses, and outline the theoretical basis for this treatment modality, including mindfulness-based cognitive therapy (MBCT). In Part II, the writers detail the therapy phases of MBCT and present a clinical case in which an advance practice psychiatric nurse integrates mindfulness-based psychotherapeutic approaches into her treatment. This clinical case may inform future clinical nursing research and advanced psychiatric nursing practice. PMID- 15529282 TI - Using mindfulness-based therapeutic interventions in psychiatric nursing practice -part II: Mindfulness-based approaches for all phases of psychotherapy--clinical case study. AB - Finding effective nursing interventions for the treatment of mental illness is a major concern for advanced practice psychiatric nurses (APPN). Increasingly, innovative psychotherapeutic treatment modalities are being used for the treatment of individuals with serious mental illness. One such innovative approach, Mindfulness-based therapeutic interventions, has been shown to relieve distress for individuals with medical and psychiatric illnesses. In Part I of this two-part article, the authors described principle concepts of mindfulness, reviewed current research in the usefulness of mindfulness practice for treatment of psychiatric illnesses. The structure and content of mindfulness-based cognitive therapy (MBCT) was also outlined and provided the theoretical basis for this treatment approach. This alternative therapeutic approach offers potential usefulness for the treatment of individuals suffering from mental illness. Part II of this article presents a more detailed explanation of mindfulness-based therapeutic approaches for all phases of psychotherapy and identifies relevant research questions that will provide an empirical base to guide clinical practice. A case study illustrates mindfulness-based therapeutic interventions. PMID- 15529283 TI - The stigma of seeking care and depression. AB - This study investigates the relationship between stigma and care-seeking for depression. One hundred-seventeen people in a primary care facility and an urban public health clinic completed the Attitudes Toward Seeking Professional Help Scale and the Attribution Questionnaire. The belief that depression was under personal control was associated with less receptivity toward seeking care, and greater anger toward the person with depression. Pity was associated with seeking professional care and with a desire to help. Dangerousness was unrelated to care seeking. Gender and setting influenced the results-for men and urban respondents there was no relationship between responsibility and care-seeking. PMID- 15529284 TI - Perceived reasons for depression among low income women of Mexican descent. AB - From a larger cross-sectional study of 315 women of Mexican descent, this secondary analysis focused on short answers to open-ended questions related to reasons given by 107 women at risk for depression (>16 on CES-D) for feelings of sadness, hopelessness, or depression within the last month. Data were analyzed using grounded theory techniques. Six categories of reasons were derived from data including: (1) partner issues, (2) family issues, (3) feelings of being alone, (4) inability to provide for material needs, (5) bodily symptoms and experiences, and (6) vague nonspecific reasons. Results are useful for designing future treatment programs. PMID- 15529285 TI - Effects of music on major depression in psychiatric inpatients. AB - The study was to assess the effectiveness of soft music for treatment of major depressive disorder inpatients in Kaohsiung City, Taiwan. A pretest-posttest with a two-group repeated measures design was used. Patients with major depressive disorder were recruited through referred by the psychiatric physicians. Subjects listened to their choice of music for 2 weeks. Depression was measured with the Zung's Depression Scale before the study and at two weekly posttests. Using repeated measures ANCOVA, music resulted in significantly better depressive scores, as well as significantly better subscores of depression compared with controls. Depression improved weekly, indicating a cumulative dose effect. The findings provide evidence for psychiatric nurses to use soft music as an empirically based intervention for depressed inpatients. PMID- 15529288 TI - Abnormal placentation and the syndrome of preeclampsia. AB - Preeclampsia, particularly the severe cases that occur early in pregnancy, is associated with defects in the (placental) cytotrophoblast differentiation pathway that leads to uterine invasion. At a morphologic level, interstitial invasion often is shallow. Perhaps more significantly, endovascular invasion, particularly the arterial component, is rudimentary. The latter defect is thought to lead to hypoperfusion of the placenta. At a molecular level, these defects are associated with particular deficits in the differentiation process whereby cytotrophoblasts--epithelial cells of ectodermal origin--assume vascular-like properties. Until recently, the question was how the latter defects could lead to the maternal symptoms of this condition. Now a possible link in the form of preeclampsia-associated changes in placental production of vasculogenic/angiogenic substances and their inhibitors has been discovered. It is likely that this new paradigm will improve both diagnosis and treatment of this life-threatening pregnancy complication. PMID- 15529289 TI - Angiogenic imbalance in the pathophysiology of preeclampsia: newer insights. AB - Angiogenesis is the process of neovascularization from preexisting blood vessels, whereas vasculogenesis is the process of blood vessel generation from angioblast precursor cells. The human placenta undergoes high levels of angiogenesis and vasculogenesis during fetal development. Additionally, the placenta undergoes a process of vascular mimicry (also referred to as pseudovasculogenesis ) in which the placental cytotrophoblasts convert from an epithelial to an endothelial phenotype during normal fetal development. Failure of placental angiogenesis and pseudovasculogenesis during placental development has been linked to the pathogenesis of preeclampsia. It currently is believed that soluble factors released by the diseased placenta lead to clinical findings of preeclampsia. This article discusses placental vascular development in health and in disease, with a focus on accumulating recent evidence that the maternal clinical syndrome of preeclampsia is an antiangiogenic state resulting from an excess of anti endothelial factors liberated by the diseased placenta. PMID- 15529290 TI - Antioxidant therapy to prevent preeclampsia. AB - Preeclampsia is a 2-stage disorder. Stage 1 is decreased placental perfusion and generates stage 2 of the disorder, the maternal syndrome characteristic of preeclampsia. How the 2 stages are linked has been a topic of intense investigation for many years. One candidate phenomenon, which includes many other suggested linkages, is oxidative stress. This hypothesis predicts that the administration of antioxidants would decrease oxidative stress and modify stage 2. Experience with the treatment of preeclampsia including a small trial of antioxidants in women with manifest preeclampsia makes it clear that the use of any therapy once preeclampsia is evident will not be successful. Trials evaluating prophylactic aspirin and supplemental calcium from early pregnancy suggest that therapy before evident preeclampsia may be successful in selected populations. Guided by these concepts and by experience with antioxidant therapy in other settings, 1 small study (<80 women in the treatment arm) was very encouraging with an almost two-thirds reduction of the frequency of preeclampsia in high-risk women. Antioxidants currently are being evaluated in several larger trials in the United States, Canada, Mexico, England, and in several developing nations. These studies should definitively establish the efficacy and safety of this therapy for the mother and fetus. PMID- 15529291 TI - Preeclampsia and the systemic inflammatory response. AB - Normal pregnancy is associated with a systemic inflammatory response. The response is exacerbated in preeclampsia and can account for its clinical features. Many of the physiologic changes of normal pregnancy are part of an acute-phase reaction, which is generated by an inflammatory response. The placenta is the proximal cause of these problems. There are several possible placental factors that may evoke the inflammatory responses that currently are being investigated. The special susceptibility of obese women, or those with diabetes or chronic hypertension, to preeclampsia is explained by the chronic systemic inflammatory responses that these women have. The clinical implications of these concepts are discussed. PMID- 15529292 TI - AT1 receptor agonistic antibodies, hypertension, and preeclampsia. AB - Immunologic mechanisms and putatively circulating mediators of preeclampsia are not a new idea, but are nevertheless compelling. Here we review studies relating to the role of agonistic antibodies that bind the second extracellular loop of the angiotensin II (AII) AT1 receptor in the pathogenesis as well as a pathologic phenotype of this disorder, focusing on observations in our laboratory. These agonistic autoantibodies (AT1-AA) appear with the development of preeclampsia and mostly are gone by 6 weeks after delivery. We have purified AT1-AA and have shown that they belong to the immunoglobulin (Ig)G3 subclass. We have shown their specificity by Western blotting, colocalization, and coimmunoprecipitation experiments. AT1-AA induce signaling in vascular cells and trophoblasts including activating protein-1 (AP-1) and nuclear factor kappa B (NF-kappaBeta) activation. The signaling results in tissue factor production and reactive oxygen species generation, both of which have been implicated in preeclampsia. AT1-AA also signal via the calcineurin-nuclear factor of activated T cells and contribute to plasminogen activator inhibitor-1 (PAI-1) production and decreased trophoblast invasion. The role of AT1-AA in preeclampsia and other severe hypertensive conditions has not yet been proven with certainty. However, we believe the findings are compelling and warrant further study. PMID- 15529293 TI - Cardiovascular changes in preeclampsia. AB - The cardiovascular system undergoes a host of changes in association with the development of preeclampsia, which ultimately lead to the classic low cardiac output-high systemic vascular resistant state. A newer hypothesis suggests that exaggeration of the normal for pregnancy hyperdynamic, low-resistance state commencing in early gestation is responsible for the genesis of the clinically apparent vasoconstrictive disease in late pregnancy. Such events may also lead to the vascular damage that persists into later life. In preeclampsia, cardiac contractility is preserved but both steady and pulsatile arterial load are increased inappropriately, failing to decrease as would occur in normal pregnancy, involving both conduit and small vessels. Abnormal adaptive mechanisms may be secondary to changes in vascular tone or vascular wall elements, and may have future implications for a woman later in life. PMID- 15529294 TI - The renal response to preeclampsia. AB - Proteinuria and decreased renal function are classic hallmarks of preeclampsia. The kidney, with its reliance on glomerular blood flow and glomerular barrier integrity, provides a unique window to view the preeclamptic disease process. This review briefly details the characteristic renal structural changes seen in preeclampsia and then focuses on the disordered renal hemodynamics and other determinants of ultrafiltration. Both renal blood flow and glomerular filtration rate (GFR) decrease in preeclampsia, although absolute values may remain above the nonpregnant range. A decrease in the ultrafiltration coefficient (K f ), in the order of 50%, either alone or in combination with reduced renal blood flow, is presented as the most likely mechanism for the decrease in GFR. Proteinuria develops, at least in part, secondary to impaired glomerular barrier integrity with a loss of size selectivity revealed by fractional dextran clearance studies and it is proposed, although yet to be proven, that this is accompanied by a loss of glomerular barrier charge selectivity. PMID- 15529296 TI - Strategies to prevent and treat preeclampsia: evidence from randomized controlled trials. AB - Preeclampsia-eclampsia is a major cause of morbidity and mortality in mothers, fetuses, and neonates worldwide, most devastating in developing nations. Its cause is still uncertain, and many controversies exist concerning its management. The World Health Organization is aware of this and is coordinating a series of systematic reviews that focus on the etiology and the best strategies for the screening, prevention, and treatment of preeclampsia. This article summarizes results from systematic reviews of randomized trials to prevent and manage preeclampsia. There is a prophylactic role of modest magnitude for low-dose aspirin but the number to treat (90 women) to avoid one case of preeclampsia still is considered high. Antioxidant and calcium supplement trials remain to be completed before firm conclusions can be rendered on their efficacy for prevention. Magnesium sulfate is effective in preventing and treating eclampsia, while severe hypertension (with or without proteinuria) requires drug therapy, but there appears to be no benefits to treating mild to moderate hypertension without proteinuria in pregnancy. Finally, our review focuses on the quality of data reviewed, suggesting the need for better evidence, and discusses the use of systematic reviews as a strategy to focus future research on this important area of reproductive medicine. PMID- 15529297 TI - Antihypertensive therapy in pregnancy. AB - The benefits of antihypertensive therapy in pregnancy remain uncertain. Blood pressure control to prevent or correct severe hypertension can avert maternal target organ damage and may allow obstetricians to prolong pregnancy or avoid hospitalization. Several factors limit the conclusions derived from systematic review of the available studies, including failure to distinguish among women with preeclampsia, gestational hypertension, or whose hypertension antedated pregnancy. As well, the application of consensus guidelines is limited by the unfortunate tendency to measure blood pressure by use of oscillometric devices rather than auscultation. We review the basis for using specific antihypertensive drugs in pregnancy and highlight important shortcomings in therapeutic knowledge that should be addressed in future studies. PMID- 15529295 TI - Animal models of preeclampsia. AB - There have been many attempts to produce animal models that mimic the hypertensive disorders of pregnancy, especially preeclampsia, but most are incomplete when compared to the full spectrum of the human disease. This review assesses a number of these models, organized according to the investigators attempt to focus on a specific pathogenic mechanism believed to play a role in the human disease. These mechanisms include uterine ischemia, impairments in the nitric oxide system, insulin resistance, overactivity of the autonomic nervous and/or renin-angiotensin systems, activation of a systemic inflammatory response, and most recently, activation of circulating proteins that interfere with angiogenesis. In addition a model of renal disease that mimics superimposed preeclampsia is discussed. Defining these animal models should help in our quest to understand the cause, as well as to test preventative and therapeutic strategies in the management of these hypertensive disorders of pregnancy. PMID- 15529300 TI - Unprecedented improvements in short-term heart rate variability due to Thought Field Therapy: response to the Pignotti retraction. AB - Short-term heart rate variability (S-HRV) is a valid and reliable measure of health and therapy effectiveness. Pignotti makes critical assertions about S-HRV with no evidence or authoritative support for these curious assertions. Short term measures have advantages over long-term measures. The power of Thought Field Therapy (TFT) is demonstrated through unprecedented HRV improvements as well as other physiologic indices such as skin color, microscopic measures, and increases in T-cell numbers after successful TFT. Bilchick and associates' hypothesis (2002), "Each increase of 10 ms in standard deviation of normal-to-normal (SDNN) conferred a 20% decrease in risk of mortality (p = .0001)," is a bold and speculative position that appears to have general, if not highly specific, merit when applied to all groups investigated with HRV. PMID- 15529299 TI - Vitamin E intake and risk of amyotrophic lateral sclerosis. AB - Oxidative stress may contribute to the pathogenesis of amyotrophic lateral sclerosis (ALS). We therefore examined prospectively whether individuals who regularly use supplements of the antioxidant vitamins E and C have a lower risk of ALS than nonusers. The study population comprised 957,740 individuals 30 years of age or older participating in the American Cancer Society's Cancer Prevention Study II. Information on vitamin use was collected at time of recruitment in 1982; participants then were followed up for ALS deaths from 1989 through 1998 via linkage with the National Death Index. During the follow-up, we documented 525 deaths from ALS. Regular use of vitamin E supplements was associated with a lower risk of dying of ALS. The age- and smoking-adjusted relative risk was 0.99 (95% confidence interval [CI], 0.69-1.41) among occasional users, 0.59 (95% CI, 0.36-0.96) in regular users for less than 10 years, and 0.38 (95% CI, 0.16-0.92) in regular users for 10 years or more as compared with nonusers of vitamin E (p for trend = 0.004). In contrast, no significant associations were found for use of vitamin C or multivitamins. These results suggest that vitamin E supplementation could have a role in ALS prevention. PMID- 15529303 TI - Callahan fails to meet the burden of proof for Thought Field Therapy claims. AB - Callahan's response evades the key issues raised by merely restating and elaborating upon what has already been said, providing citations that are out of context and irrelevant to the issues at hand, and misrepresenting what was actually said by his critics and me and the authors of articles he cites. He spends paragraphs refuting "straw men." He provides additional anecdotes, which offer no convincing evidence for his claims. His critics have expressed concern that Callahan and Thought Field Therapy (TFT) proponents will cite his response article, as published in the Journal of Clinical Psychology, to promote TFT, as TFT proponents have repeatedly done for the non-peer-reviewed earlier issue devoted to TFT. Callahan has been given an unprecedented opportunity to present his work in a reputable journal without prior peer review and has failed to meet the burden of proof for his claims, thus undermining his own claim, that his work has been rejected solely as a result of bias against innovation. PMID- 15529301 TI - An empirical investigation of Albert Ellis's binary model of distress. AB - In the current literature, distress is typically described according to a unitary model: High levels of distress are conceptualized as a high level of negative affect while low levels of distress are typically conceptualized as a low level of negative affect. On the other hand, Albert Ellis (1994) and some of his rational-emotive and cognitive-behavioral professional colleagues have more recently described distress as a binary construct composed of two different components: functional negative feelings (e.g., sad) and dysfunctional negative feelings (e.g., worthless). In two studies using 55 U.S. breast-cancer patients and 45 Romanian breast-cancer patients, respectively, we compared hypotheses derived from unitary and binary models of distress. The results revealed that in a stressful situation (i.e., upcoming breast surgery) high levels of irrational beliefs were associated with a high level of both functional and dysfunctional negative feelings while low levels of irrational beliefs were associated with a low level of dysfunctional negative feelings and a high level of functional negative feelings. Thus, that support for the binary model of distress found in both U.S. and Romanian samples suggests both the robustness and the generalizability of the results. Theoretical and practical implications are discussed. PMID- 15529304 TI - Coverage of soft-tissue defects of the hand with free fascial flaps. AB - Coverage of exposed functional structures such as tendons, bones, vessels, or nerves at the dorsal and palmar surface of the hand requires thin, supple tissue to provide adequate range of motion and a satisfying aesthetic result. The purpose of this retrospective study was to evaluate the functional and aesthetic results after coverage of the hand with free fascial flaps. From 1994-2002, 14 patients underwent free fascial flap coverage of the hand with 4 tempo-parietal fascia flaps and 11 serratus fascia flaps. Eight patients could be reexamined and answered a questionnaire about their satisfaction with the functional and aesthetic results. The mean follow-up was 41.7 months. Average active range of motion of the hand, functional improvement, and the aesthetic result were satisfying in all follow-up patients. No secondary debulking or other contouring procedures were required. We recommend the use of free fascial flaps as a valuable alternative to fasciocutaneous or muscle flaps, since the functional results are excellent, no additional procedures were necessary, and the aesthetic results are appealing. PMID- 15529305 TI - Sex and tremor location: similarities between essential tremor and cervical dystonia. PMID- 15529306 TI - Diagnostic procedures after a positive spiral computed tomography lung carcinoma screen. AB - BACKGROUND: Low-radiation dose spiral computed tomography (LDCT) currently is being evaluated as a screening modality for lung carcinoma in a randomized trial. Although several diagnostic algorithms for the workup of positive LDCT screens have been proposed, to the authors' knowledge there is no widely accepted standard to date and there are few nationwide data concerning how such diagnostic workups are actually being performed outside a research protocol setting. METHODS: The Lung Screening Study (LSS) was a multicenter feasibility trial that randomized 1660 subjects to undergo LDCT and an equivalent number to undergo chest X-ray. Subjects with positive screens were referred to their own health care providers for diagnostic follow-up; LSS did not specify a diagnostic algorithm. LSS collected and abstracted medical records regarding procedures employed in the diagnostic workup of positive screens. RESULTS: Of the 522 subjects with a positive LDCT screen at baseline or at Year One, 12% underwent biopsy. Biopsy was less likely to be performed in subjects with 4-9-mm nodules (5%) than in subjects with nodules measuring 10+ mm (25%) or in subjects with no nodules but other suspicious findings (15%). Among 63% of the subjects who underwent chest CT on follow-up, the median time between screening and first follow-up chest CT was 82 days. Only a minority of subjects received diagnostic workups that were consistent with published algorithms. CONCLUSIONS: The data from the current study represent the experience of subjects followed by their health care providers in five different U.S. metropolitan areas and one rural area. As such, they provide some indication of practices in the U.S. with regard to the diagnostic workup of patients with positive spiral CT screens. PMID- 15529307 TI - Opioid switching from transdermal fentanyl to oral methadone in patients with cancer pain. AB - BACKGROUND: Patients with cancer often are rotated from other opioids to methadone to improve the balance between analgesia and side effects. To the authors' knowledge, no clear guidelines currently exist for the safe and effective rotation from transdermal fentanyl to methadone. METHODS: The authors evaluated a protocol for switching opioid from transdermal fentanyl to oral methadone in 17 patients with cancer. Reasons for switching were uncontrolled pain (41.1% of patients) and neurotoxic side effects (58.9% of patients). Methadone was initiated 8-24 hours after fentanyl withdrawal, depending on the patient's previous opioid doses (from < 100 microg per hour to > 300 microg per hour). The starting methadone dose was calculated according to a 2-step conversion between transdermal fentanyl:oral morphine (1:100 ratio) and oral morphine:oral methadone (5:1 ratio or 10:1 ratio). The correlation between previous fentanyl dose and the final methadone dose or the fentanyl:methadone dose ratio was assessed by means of Pearson and Spearman correlation coefficients (r), respectively. A Friedman test was used to compare pain intensity before and after the switch and the use of daily rescue doses. RESULTS: Opioid rotation was fully or partially effective in 80% and 20%, respectively, of patients with somatic pain. Neuropathic pain was not affected by opioid switching. Delirium and myoclonus were reverted in 80% and 100% of patients, respectively, after opioid switching. A positive linear correlation was obtained between the fentanyl and methadone doses (Pearson r, 0.851). Previous fentanyl doses were not correlated with the final fentanyl:methadone dose ratios (Spearman r, - 0.327). CONCLUSIONS: The protocol studied provided a safe approach for switching from transdermal fentanyl to oral methadone, improving the balance between analgesia and side effects in patients with cancer. PMID- 15529309 TI - Impact of high-dose granulocyte transfusions in patients with cancer with candidemia: retrospective case-control analysis of 491 episodes of Candida species bloodstream infections. AB - BACKGROUND: The efficacy and feasibility of donor granulocyte transfusion therapy (GTX) have changed considerably over the past four decades. The authors sought to determine the impact of high-dose (approximately 5.5 x 10(10) cells) GTX in patients with candidemia. METHODS: The authors' case-control retrospective analysis comprised 491 consecutive patients treated at The University of Texas M. D. Anderson Cancer Center (Houston,TX) from 1993 to 2000. The cohort included 29 patients with Candida species bloodstream infection who had received GTX and 462 who had not. RESULTS: Both groups were comparable in age, gender, APACHE II score, recent chemotherapy received, broad-spectrum antibiotics, systemic corticosteroids, radiotherapy, intravascular catheter, and concordant antifungal therapy (P > or = 0.1). The patients who received GTX compared with those who did not had a higher incidence of underlying leukemia (86% vs. 29%, P <0.001), persistent neutropenia (59% vs. 18%, P <0.001), non-Candida albicans candidemia (Candida glabrata, 35%; Candida krusei, 31%: 90% vs. 67%, P=0.01), and breakthrough invasive mycosis (62% vs. 23%, P <0.001). Neutropenia was more prolonged in patients who received GTX (28 vs. 10 days, P <0.001). Also, more of the patients who received GTX had received hematopoietic stem cell transplantations (28% vs. 13%, P = 0.03), exposure (within 4 weeks) to antifungals (79% vs. 38%, P <0.001), and stays in critical care units (62% vs. 40%, P=0.02). The overall attributable mortality rate for 25 evaluable recipients of GTX was 48% (n=12), compared with 45% (n=115) of 254 evaluable patients in the control group (P=0.5). Of the 158 patients with leukemia, 25 (16%) had received GTX. In patients with leukemia, more of those who had received GTX experienced disseminated candidiasis (44% vs. 26%; P <0.07) and persistent neutropenia (68% vs. 43%, P <0.02), had candidemia that was more prolonged (> 72 hours, P <0.02), and had more stays in critical care units (68% vs. 44%, P <0.03). On the bases of a reduced multivariate model, a significantly increased risk of death was found for patients with hematopoietic stem cell transplantation (odds ratio [OR]=2.51; 95% confidence interval [95% CI], 0.99-6.31; P <0.05), for patients with persistent neutropenia (OR=4.57; 95% CI, 1.99-10.47; P <0.0003), and for patients with leukemia who also had prolonged candidemia (OR=3.59; 95% CI, 1.61-7.98; P <0.002), disseminated candidiasis (OR=5.19; 95% CI, 2.17-12.42; P <0.0002), or non-C. albicans candidemia (OR=5.02; 95% CI, 1.07-23.64; P <0.04). In patients with leukemia, death was attributable to candidemia in 50% of the GTX recipients, compared with 59% of the non-GTX patients who had received antifungal therapy alone (P=0.4). CONCLUSIONS: Despite the presence of multiple predictors of increased mortality, high-dose GTX therapy in these high-risk patients with cancer was associated with better than expected survival rates. PMID- 15529310 TI - The equivalence and difference between the English and Chinese versions of two major, cancer-specific, health-related quality-of-life questionnaires. AB - BACKGROUND: English and Chinese are two of the most widely used primary languages in the world. Patients in many cancer centers have a variety of ethnic backgrounds and primary languages. The comparability of version 4 of the English and Chinese versions of the Functional Assessment of Cancer Therapy-General (FACT G) and version 3 of the European Organization for the Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ-C30) have not been established. METHODS: In total, 1136 ethnic Chinese patients with cancer were recruited from the National Cancer Centre of Singapore. Patients chose to answer an English or Chinese questionnaire, according to their own preference. Multiple regression analysis was used to adjust for differences in demographic and health characteristics. Equivalence was confirmed if the 90% confidence intervals of the adjusted mean difference fell completely within an equivalence zone of +/- 0.25 standard deviations (SD). RESULTS: The English and Chinese versions of the Total, Emotional, and Functional Well Being Scales of the FACT-G and the Physical and Emotional Functioning Scales of the EORTC QLQ-C30 were equivalent. Scores for the other scales on the two questionnaires, at most, had a small differences that did not exceed 0.5 SD. Nevertheless, the Chinese translation of the question "I have a lack of energy" in the Physical Well Being Scale of the FACT-G produced results that differed from the results produced by the original English version. CONCLUSIONS: Data collected from English-speaking and Chinese-speaking respondents were capable of being pooled, and either version could be used for bilingual respondents. Nevertheless, the authors recommend modification of the Physical Well Being question that produced different results ("I have a lack of energy"). PMID- 15529311 TI - Immunochemotherapy with rituximab and temozolomide for central nervous system lymphomas. PMID- 15529312 TI - Heterogeneity of risk for melanoma and pancreatic and digestive malignancies: a melanoma case-control study. AB - BACKGROUND: Data addressing the interfamilial heterogeneity of melanoma are limited. In the current study, the authors assessed melanoma risk according to family history of melanoma and other melanoma-associated malignancies and evaluated the familial heterogeneity of melanomas, pancreatic malignancies, and gastrointestinal malignancies. METHODS: The authors obtained patient histories of malignancy in first-degree relatives as part of a clinic-based case-control study. The case group included 737 newly diagnosed patients with invasive melanoma, and the control group included 1021 outpatients from clinics at the same medical centers. To assess heterogeneity of risk among families affected by melanoma, a nonparametric method was used to detect extrabinomial variation. In addition, selected patients with melanoma (n=133) were tested for germline mutations in CDKN2A. RESULTS: The adjusted odds ratio associated with a family history of melanoma was 1.7 (95% confidence interval, 1.1-2.7). Family histories of pancreatic, gastrointestinal, brain, breast, or lymphoproliferative disease did not increase the risk of melanoma significantly. Among case families, significant evidence of familial heterogeneity was found for melanomas, but not for pancreatic or gastrointestinal malignancies. Two mutations in CDKN2A previously associated with melanoma risk were identified among the 133 patients tested in the case group; mutation detection did not differ between families with low and high heterogeneity scores. CONCLUSIONS: Familial heterogeneity testing in the study population did not improve the selection of high-risk families for genetic study. Even in a large case-control study, few families that had multiple members with melanoma were identified, and family members with pancreatic malignancies were rare. PMID- 15529313 TI - 2-Chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU) inhibits prostate carcinoma cell growth via p53-dependent and p53-independent pathways. AB - BACKGROUND: Prostate carcinoma is the most commonly occurring malignancy in men. Although 2-chloroethyl-3-sarcosinamide-1-nitrosourea (SarCNU), an analog of the chloroethylnitrosoureas, has been used in the treatment of advanced solid tumors, the molecular mechanisms underlying the antineoplastic activity of this agent are not well understood. In the current study, the authors sought to investigate the effects of SarCNU on prostate carcinoma cell growth in vivo and in vitro. METHODS: Male SCID mice underwent subcutaneous implantation (on both flanks) of human CWR-22 and CWR-22R prostate carcinoma xenografts. Mice were treated with either vehicle or 60 or 80 mg SarCNU per kg body weight for 5 days, with tumor growth being assessed every 3 days. Animals were sacrificed 21 days after the final injection, and tumors subsequently were collected, weighed, and processed for analysis. Immunohistochemical analyses were performed to obtain data on the localization of p53 and p21Cip1/Waf1. Cell counting, terminal deoxynucleotidyltransferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) assays, cell cycle analyses, Western blotting, and in vitro kinase assays were performed to determine the effects of SarCNU on growth, apoptosis, cell cycle arrest, cell cycle-regulated protein levels, and Cdc-2 activity, respectively. RESULTS: SarCNU reduced tumor incidence and inhibited the growth of CWR-22 and CWR-22R xenografts. In addition, treatment with this agent led to increases in p21Cip1/Waf1 levels and p53 phosphorylation at Ser15. In vitro administration of SarCNU to cells with wild-type p53 (LNCaP and primary CWR-22 cells) and cells with mutant p53 (PC-3 cells) resulted in G2/M arrest and the reduction of cellular Cdc-2 activity. Up-regulation of p53 levels, p53 phosphorylation at Ser15, and p21Cip1/Waf1 levels in primary CWR-22 and LNCaP cells, as well as up-regulation of Cdc-2 phosphorylation at Tyr15 in PC-3 cells, was detected. CONCLUSIONS: SarCNU induced G2/M arrest in prostate carcinoma cells via p53-dependent up-regulation of p21Cip1/Waf1 and p53-independent phosphorylation of Cdc-2 at Tyr15. These findings suggest a potential role for SarCNU in the treatment of prostate malignancies. PMID- 15529314 TI - Adhesion of human U937 macrophages to phosphorylcholine-coated surfaces. AB - A new type of amphiphilic phosphorylcholine (PC) polymer was used in this work to develop a cell culture surface that allows the attachment of U937 macrophages. The PC polymer was a random copolymer of N-isopropylacrylamide (45%), N (phosphorylcholine)-N'-(ethylenedioxy-bis(ethyl)) acrylamide (41%), and the hydrophobic monomer N-(n-octadecyl) acrylamide (14%). Polypropylene (PP) films (1 cm2) were coated with the polymer solution by immersion. U937 macrophage suspensions were applied on PC polymer-coated surfaces and incubated for up to 72 h at 37 degrees C. While U937 cells did not adhere to PP, ammonia, nitrogen, or oxygen plasma-treated surfaces, they attached rapidly on PC-coated surfaces (< 1 h), proliferated, and stayed attached to the modified surface for at least 72 h, suggesting that unique features of the PC polymer, and the U937 macrophages, are responsible for the attachment of these cells. We compared the effect of Co2+ and Cr3+ ions on the expression of bone-resorbing cytokines (TNF-alpha, IL-6, IL 1beta) in U937 macrophages cultured on PC-coated surfaces to the response of U937 macrophages in suspension. Cytokine gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Addition of Co2+ and Cr3+ ions led to a significant increased expression of TNF-alpha in both cultured and suspension cells. On the other hand, Co2+ and Cr3+ ions had a weak stimulatory effect or no effect on IL-1beta and IL-6, respectively, in both cultured and suspension cells. In conclusion, the use of PC polymer-modified surfaces might offer promising new opportunities for the culture of human U937 cells and may also point to the mechanism by which macrophages interact with lipid bilayers of biological membranes. PMID- 15529316 TI - Parapharyngeal lymph node metastasis: an unusual presentation of papillary thyroid carcinoma. PMID- 15529317 TI - Three new families with arterial tortuosity syndrome. AB - Arterial tortuosity syndrome (ATS) is a rare condition with autosomal recessive inheritance characterized by connective tissue abnormalities. The most specific clinical findings are cardiovascular anomalies including tortuosity, lengthening, aneurysm, and stenosis formation of major arteries. Also ventricular hypertrophy is frequently present. Other anomalies are skin hyperextensibility and cutis laxa, joint laxity or contractures of the joints, and inguinal herniae. Histology shows disruption of elastic fibers of the media. These features suggest that ATS is a connective tissue disorder. A biochemical or molecular defect has not yet been identified. We describe here nine additional ATS patients from three consanguineous Moroccan families and review a total of 35 patients with this uncommon condition. PMID- 15529318 TI - Evolving treatment strategies for parameningeal rhabdomyosarcoma: the experience of the Istituto Nazionale Tumori of Milan. AB - BACKGROUND: Ninety pediatric patients with parameningeal rhabdomyosarcoma (PM RMS) were treated between 1970 and 2002 at the Istituto Nazionale Tumori, Milan. METHODS: Combined treatment was implemented in high-risk patients with whole brain irradiation and intrathecal chemotherapy between 1978 and 1981 and intrathecal methotrexate alone between 1982 and 1987. Radiotherapy was given with conventional fractionation until 1988 and hyperfractionated accelerated radiotherapy (HART) thereafter. RESULTS: The 5-year disease-free survival was 41.3%, increasing from 15.4% (1970-1977) to 55.9% (1989-2002) for localized cases. Local failure rates were 41% and 24% with conventional fractionation and HART, respectively. In patients treated with HART, acute toxicities were more pronounced, but the treatment compliance was better. Severe radiation-related late sequelae were observed. CONCLUSIONS: The outcome for patients with PM-RMS has improved over the years but is still not as satisfactory as that for patients with RMS of other sites. Thanks to improvements in radiotherapy techniques and chemotherapy efficacy, meningeal prophylaxis is no longer necessary. Although more evidence is needed to confirm our data, the results obtained with twice daily HART seem promising. PMID- 15529319 TI - Sinonasal hemangiopericytoma-like tumor with true pericytic myoid differentiation: a clinicopathologic and immunohistochemical study of five cases. AB - BACKGROUND: Sinonasal hemangiopericytoma-like tumor (SHPCL) is an uncommon vascular tumor that is sometimes difficult to diagnose. METHOD: We report five cases arising from the nasal cavity. The tumors were subdivided into two types, prototype and soft tissue type, by the difference in histologic features. RESULTS: The prototype, made up of three tumors, is characterized by uniform, spindle-shaped myoid cells that have eosinophilic cytoplasm and uniform nuclei and lay down in an orderly fashion. The soft tissue type, made up of two tumors, is similar to the hemangiopericytoma of soft tissue. Immunohistochemically, the tumors were positive for vimentin (100%), smooth muscle actin (80%), muscle specific actin (100%), fibroblast growth factor-2 (100%), and vascular endothelial growth factor (100%). All were negative for cytokeratin, desmin, CD34, and factor VIII-related antigen. These histopathologic features suggest myoid differentiation and angiogenic activity of the tumors. All the tumors were treated with wide surgical excision. Only one case of soft tissue-type tumor with active mitoses received additional postoperative radiotherapy. No recurrence was found during 13 to 120 months of follow-up. CONCLUSIONS: Our data suggest that SHPCL with convincing myoid differentiation may have a more favorable clinical course than its soft tissue counterpart. PMID- 15529320 TI - Glossal angiomyoma: imaging findings and endovascular treatment. AB - BACKGROUND: An angiomyoma is an uncommon, benign tumor characterized by numerous vascular channels intermixed with bundles of smooth muscle cells. Oral manifestations are quite rare. We describe for the first time the CT, MRI, and angiographic imaging features and successful preoperative endovascular embolization of an angiomyoma of the tongue. The pathologic findings before and after embolization are also described. METHODS AND RESULTS: A 25-year-old man was seen with a rapidly enlarging tongue mass. Imaging studies revealed the extent and hypervascular nature of this tumor. The diagnosis of angiomyoma was confirmed by histologic examination. Preoperative embolization proved to be helpful in the surgical management of this lesion. CONCLUSIONS: Angiomyoma should be considered in the differential diagnosis of any well-circumscribed, hypervascular, soft tissue tumor in the mouth. In addition, endovascular embolization may be a useful adjunct that facilitates resection. PMID- 15529321 TI - Partial laryngectomy for recurrent glottic carcinoma after radiotherapy. AB - BACKGROUND: Early laryngeal cancer is treated with surgery or radiotherapy. A partial laryngectomy instead of a total laryngectomy can be used for treating patients with radiation failures. METHODS: Patients were grouped by the two types of partial laryngectomies we performed: group I, endoscopic laser surgery (n = 42); and group II, frontolateral partial laryngectomy (n = 21). RESULTS: With CO2 laser treatment, 14 of 24 patients (no involvement of the anterior commissure) and eight of 18 patients (involvement of the anterior commissure) were cured. With the frontolateral partial laryngectomy, we achieved local control in 15 of 21 patients. CONCLUSIONS: If the surgeon is familiar with the different techniques of, and indications for, partial laryngectomy, this can be a good and satisfying treatment in selected patients with radiation failure for glottic cancer PMID- 15529322 TI - History of environmental and molecular mutagenesis. AB - Environmental and Molecular Mutagenesis, the journal of the Environmental Mutagen Society (EMS), marks its 25th anniversary in 2004. The journal, originally called Environmental Mutagenesis, was established in 1979 with Seymour Abrahamson as its first editor. The development of the journal is closely linked to the evolution of the fields of mutation research and genetic toxicology. This perspective traces the founding of the journal and discusses its editorial history, growth, content, style, administration, and relationship to the EMS. PMID- 15529323 TI - Antimutagenic activity of spearmint. AB - The antimutagenic activity of spearmint (Mentha spicata), a popular food flavoring agent, was studied in the Salmonella assay. Spearmint leaves were brewed in hot water for 5 min at concentrations up to 5% (w/v), and the water extracts were tested against the direct-acting mutagens 4-nitro-1,2 phenylenediamine (NPD) and 2-hydroxyamino-3-methyl-3H-imidazo[4,5-f]quinoline (N OH-IQ) using Salmonella typhimurium strain TA98. Nontoxic concentrations of spearmint extract inhibited the mutagenic activity of N-OH-IQ in a concentration dependent fashion, but had no effect against NPD. These experiments by design focused on the water extract consumed commonly as an herbal tea, but chloroform and methanol extracts of spearmint also possessed antimutagenic activity against N-OH-IQ. Water extract of spearmint inhibited the mutagenic activity of the parent compound, 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ), in the presence of rat liver S9; however, the concentration for 50% inhibition (IC50) against IQ was approximately 10-fold higher than in assays with N-OH-IQ minus S9. At concentrations similar to those used in the Salmonella assays, spearmint extract inhibited two of the major enzymes that play a role in the metabolic activation of IQ, namely, cytochromes P4501A1 and 1A2, based on ethoxyresorufin O deethylase and methoxyresorufin O-demethylase assays in vitro. In vivo, rats were given spearmint water extract (2%; w/v) as the sole source of drinking fluid before, during, and after 2-week treatment with IQ; colonic aberrant crypt foci were inhibited significantly at 8 weeks (P < 0.05, compared with rats given IQ alone). Collectively, these findings suggest that spearmint tea protects against IQ and possibly other heterocyclic amines through inhibition of carcinogen activation and via direct effects on the activated metabolite(s). PMID- 15529324 TI - History and rationale of genetic toxicity testing: an impersonal, and sometimes personal, view. AB - Genetic toxicity testing is a necessary and pivotal component of product development and registration. This article traces the historical development and evolution of genetic toxicity testing, and the rationale for such testing, and identifies some of the individuals who played key roles in this process. The evolution of the present test batteries and some of the research and rationales behind the decisions to accept or reject tests are described. PMID- 15529325 TI - History of the science of mutagenesis from a personal perspective. AB - A career in the study of mutagenesis spanning 50 years is a gift few scientists have been bestowed. My tenure in the field started in 1953, the year the structure of DNA became known (Watson and Crick [1953]: Nature 171:737). Before that time, it was suspected that DNA was the genetic material based on the research of Oswald T. Avery (Avery et al. [1944]: J Exp Med 79:137), but many scientists still believed that proteins or polysaccharides could be the genetic material. The present article describes a lifetime of personal experience in the field of chemical mutagenesis. The methods used to treat viruses with chemical mutagens were well developed in the 1950s. Here I review the early use of nitrous acid and hydroxylamine as mutagens in eukaryotes, the development of methods for the metabolic activation of mutagens by microsomal preparations, and the selection of a mutant tester set for the qualitative characterization of the mutagenic activity of chemicals. These studies provided critical background information that was used by Bruce Ames in the development of his Salmonella/microsome assay, widely known as the Ames test (Ames et al. [1973]: Proc Nat Acad Sci USA 70:2281-2285). This article also describes how a set of diagnostic chemical mutagens was selected and used to identify the molecular nature of gene mutations. Today, DNA sequencing has replaced the use of diagnostic mutagens, but studies of this kind formed the foundation of modern mutation research. They also helped set the stage for the organization of the Environmental Mutagen Society and the Environmental Mutagen Information Center, which are described. The article ends with the development of mammalian single cell mutation assays, the first system for studying in vivo mutagenesis using recoverable vectors in transgenic animals, other mutation assays in intact mammals, and my thoughts on the critically important area of germ cell mutagenesis. This narrative is not a complete autobiographical account, in that I have selected only those experiences that I feel are important for the history of the field and the edification of today's students. I hope I have shown that science not only is a valuable pursuit but can also be fun, stimulating, and satisfying. A good sense of humor and the knowledge that many discoveries come by serendipity are essential. PMID- 15529326 TI - Regarding the October 2001 Journal of Clinical Psychology special issue on Thought Field Therapy: retraction of conclusions in the article "heart rate variability as an outcome measure for Thought Field Therapy in clinical practice". AB - This article is a retraction of the conclusions drawn in a previous article, published as part of a special October 2001 issue of the Journal of Clinical Psychology on Thought Field Therapy (TFT). I decided to write this retraction after reconsidering a number of issues raised in the critiques of the articles. Additionally, subsequent misinterpretations of the literature on heart rate variability (HRV) by Roger Callahan, which led to further questioning of his premises and claims regarding TFT and HRV as represented in the articles, are discussed. I conclude that the burden of proof is on TFT proponents to demonstrate its efficacy and that well-designed controlled studies using standardized assessment measures and long-term follow-up must be performed to allow the scientific community to take claims made for TFT seriously. PMID- 15529328 TI - The MPSim-Dock hierarchical docking algorithm: application to the eight trypsin inhibitor cocrystals. AB - To help improve the accuracy of protein-ligand docking as a useful tool for drug discovery, we developed MPSim-Dock, which ensures a comprehensive sampling of diverse families of ligand conformations in the binding region followed by an enrichment of the good energy scoring families so that the energy scores of the sampled conformations can be reliably used to select the best conformation of the ligand. This combines elements of DOCK4.0 with molecular dynamics (MD) methods available in the software, MPSim. We test here the efficacy of MPSim-Dock to predict the 64 protein-ligand combinations formed by starting with eight trypsin cocrystals, and crossdocking the other seven ligands to each protein conformation. We consider this as a model for how well the method would work for one given target protein structure. Using as a criterion that the structures within 2 kcal/mol of the top scoring include a conformation within a coordinate root mean square (CRMS) of 1 A of the crystal structure, we find that 100% of the 64 cases are predicted correctly. This indicates that MPSim-Dock can be used reliably to identify strongly binding ligands, making it useful for virtual ligand screening. PMID- 15529327 TI - Static mechanical properties of hydroxyapatite (HA) powder-filled acrylic bone cements: effect of type of HA powder. AB - This work reports on the effect of the amount (0, 10, and 30 wt %) and type of HA powder incorporated into an acrylic bone cement on the tensile properties, compression properties, and fracture toughness. The three different types of HA powders used were synthesized in the laboratory and coated with a silane agent prior to incorporation into the cement powder, and differed in particle size, water content, surface area, and crystallinity. It was found that the inclusion of any type of HA powder led to an increase in the tensile modulus (ET), but all the other mechanical properties of the cement decreased (relative to the values of the unfilled cement). The increase in ET is attributed to the good adhesion between the filler and the cement matrix, which is due to the silane coating agent. The decrease in the other mechanical properties may be a consequence of HA powder agglomeration and porosity. Hydroxyapatite morphology and crack-growth mechanisms were analyzed by scanning electronic microscopy (SEM). PMID- 15529329 TI - A scalable parallel Monte Carlo method for free energy simulations of molecular systems. AB - We present a method of parallelizing flat histogram Monte Carlo simulations, which give the free energy of a molecular system as an output. In the serial version, a constant probability distribution, as a function of any system parameter, is calculated by updating an external potential that is added to the system Hamiltonian. This external potential is related to the free energy. In the parallel implementation, the simulation is distributed on to different processors. With regular intervals the modifying potential is summed over all processors and distributed back to every processor, thus spreading the information of which parts of parameter space have been explored. This implementation is shown to decrease the execution time linearly with added number of processors. PMID- 15529330 TI - Reevaluating confined placental mosaicism. AB - Chromosomal mosaicism was found in 38 of 4,000 chorionic villus samples examined from 1998 to 2003. A small fraction of these (5/38) were confirmed as true mosaics by analysis of amniotic fluid. Twenty-nine cases that fit the definition of confined placental mosaicism were followed with clinical and cytogenetic analysis throughout the pregnancy, at birth and in a few cases into infancy. This was done to determine the prognostic interpretation of prenatal cytogenetic results from multiple specimens in a single pregnancy and thus allow for reevaluation of the genetic counseling. In 2 of these 29 cases, low-level mosaicism was found in the neonate, and in 1 of these the chromosome abnormality is probably the cause of the resulting minor phenotypic abnormalities. Families face unique difficulties when confined placental mosaicism is the prenatal diagnosis, and it is extremely important that the counseling they receive takes into consideration the unlikely possibility of the placental abnormality appearing in fetal tissues. PMID- 15529331 TI - Novel polyelectrolyte carboxymethyl konjac glucomannan-chitosan nanoparticles for drug delivery. II. Release of albumin in vitro. AB - Carboxymethyl konjac glucomannan-chitosan (CKGM-CS) nanoparticles were spontaneously prepared under very mild conditions via polyelectrolyte complexation. Bovine serum albumin (BSA), as a model protein drug, was incorporated into the CKGM-CS nanoparticles. The physicochemical properties of the BSA-loaded nanoparticles were identified by Zetasizer 3000 and FTIR spectrophotometry. Their sizes were from 330 nm to 900 nm; zeta potentials were positive according to varies CKGM/CS ratios. The encapsulation efficiency was up 20%. The release behavior in vitro of BSA from the nanoparticles was also investigated. We could find that the BSA release from the CKGM-CS nanoparticles is much more influenced by the CS coating layer than by the CKGM inner structure. And the CKGM-CS matrices not only exhibited pH-responsive properties, but ionic strength-sensitive properties. These systems may present a potential for pulsatile protein drug delivery. PMID- 15529332 TI - In vitro characterization of 6-[18F]fluoro-A-85380, a high-affinity ligand for alpha4beta2* nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors are involved in tobacco dependence and several other neuropathologies (e.g., Alzheimer's disease, Parkinson's disease), as well as in attention, learning, and memory. Performing in vivo imaging of these receptors in humans holds great promise for understanding their role in these conditions. Recently, three radiohalogenated analogs of 3-(2(S) azetidinylmethoxy)pyridine (A- 85380) were used successfully for the in vivo visualization of alpha4beta2* nicotinic receptors in the human brain with PET/SPECT. Herein, we present the results of the in vitro characterization of one of these radioligands, 6-[18F]fluoro-3-(2(S)-azetidinylmethoxy)-pyridine (6 [18F]fluoro-A-85380), which is a fluoro-analog of the potent nonopioid analgesic ABT-594. In human postmortem cortical tissue, 6-[18F]fluoro-A-85380 reversibly binds with high affinity to a single population of sites (Kd = 59 pM at 37 degrees C, Bmax = 0.7 pmol/g tissue). The binding is fully reversible and is characterized at 37 degrees C by T(1/2assoc) = 2.2 min (at a ligand concentration of 39 pM) and by T(1/2dissoc) = 3.6 min. 6-Fluoro-A-85380 exhibits clear selectivity for alpha4beta2* over the other major mammalian nicotinic receptor subtypes: alpha7, alpha3beta4, and muscle-type. These results suggest that 6 [18F]fluoro-A-85380 is a promising radioligand for in vivo imaging of brain alpha4beta2* nicotinic receptors. PMID- 15529333 TI - Elution from light-cured dental composites: comparison of trimethacrylate and dimethacrylate as base monomers. AB - The effect of the number of functional units on a monomer on the elution property of dental composites was first studied. Elution from a composite of a trifunctional methacrylate, 1,1,1-tris[4-(2'-hydroxy-3' methacryloyloxypropoxy)phenyl]ethane (THMPE), was compared to that of bis-GMA, a typical difunctional base monomer in current dental composites. The degrees of cure and the water solubilities of composites prepared from the two kinds of methacrylates were measured. The concentration of unreacted methacrylates (base monomer and diluent) present in the photo-cured composites as well as the quantity of the methacrylates eluted into water (or 75%, volume fraction, of ethanol in water) from the composites were determined by high-performance liquid chromatography (HPLC). THMPE-based composites showed lesser amounts of residual and eluted methacrylates when compared with bis-GMA composites. This is attributed to the higher degree of functionality and larger molecular size of THMPE compared with those of bis-GMA. PMID- 15529334 TI - Effects of repeated cocaine on the release and clearance of dopamine within the rat medial prefrontal cortex. AB - Previous data suggest that cocaine-induced dopamine (DA) transmission within the medial prefrontal cortex (mPFC) undergoes time-dependent changes during withdrawal from repeated cocaine administration. The current studies assessed two potential mechanisms that may underlie this neuroadaptation. One set of experiments examined alterations in DA clearance in the mPFC of rats that had been pretreated with four administrations of cocaine (15 mg/kg, i.p.; once per day for 4 days) and were withdrawn 1, 7, or 30 days. No significant changes in mPFC DA uptake into crude mPFC synaptosomes or in mPFC DA transporter levels were observed at any of the time points examined. Uptake assay and Western blotting sensitivity was confirmed with prefrontal 6-hydroxydopamine lesions, which significantly reduced [3H]DA uptake and DA transporter immunoreactivity in mPFC synaptosomes. To evaluate temporal changes in DA release resulting from repeated cocaine, additional experiments utilized in vivo microdialysis to locally infuse KCl (10, 30, or 100 mM) into the mPFC over the same withdrawal time course used in the uptake studies. After 1-7 days of withdrawal, KCl-stimulated DA release was significantly reduced in the mPFC of cocaine-pretreated animals. However, after 30 days of withdrawal the evoked release of DA in the mPFC of saline- and cocaine-pretreated animals was similar. These data suggest that previously reported modulation of cocaine-induced mPFC DA transmission occurring upon withdrawal from repeated cocaine might arise from transient changes in DA releasability rather than clearance. The relevance of these findings is discussed in relation to mPFC involvement in psychostimulant sensitization. PMID- 15529335 TI - Mapping interactions between dopamine and adenosine A2a receptors using pharmacologic MRI. AB - Adenosine receptors in the basal ganglia are implicated in regulation of dopamine function and release. We investigated the interactions between dopamine receptors and adenosine receptors in the basal ganglia using pharmacologic MRI (phMRI) in rats. Stimulation of dopamine receptors was achieved using administration of 2 mg/kg of amphetamine. Then we investigated the antagonism of these changes using the selective A2a receptor antagonist 3,7-dimethyl-1-propargylaxanthine (DMPX). Amphetamine alone caused large increases (10-30%) in relative cerebral blood volume (rCBV) in caudate/putamen (CPu), nucleus accumbens (NAcc), thalamus, and frontal and cingulate cortices with changes that persisted for 70-80 min. DMPX alone (5 mg/kg) induced decreases in rCBV (approximately 8-10%) in NAcc, CPu, and olfactory tubercule, with smaller changes in thalamus (-6%) consistent with the regional distribution of A2a receptors. We examined the interactions between amphetamine and DMPX by assessing the effects of DMPX (5 mg/kg) administration 20 min after injection of 3 mg/kg amphetamine. These experiments showed that DMPX immediately decreased the rCBV increase induced by amphetamine in NAcc, CPu, and thalamus but not in cingulate or sensorimotor cortex. Companion microdialysis experiments showed that dopamine release in CPu was decreased in a similar manner. These experiments demonstrate the utility of phMRI for probing, in a noninvasive manner, the temporal and spatial dynamics of neurotransmitter interactions. PMID- 15529336 TI - Synthesis and biodistribution of [18F]FE@CIT, a new potential tracer for the dopamine transporter. AB - In the last decade radiolabeled tropane analogs based on beta-CIT have proven indispensable for the imaging of the dopamine transporter. However, further improvements in their pharmacodynamic and pharmacokinetic features are desirable. An important improvement, yielding in higher affinity to the dopamine transporter (DAT) vs. serotonin transporter (SERT), can be achieved by a simple replacement of the carboxylic methyl ester group in beta-CIT by a fluoroethyl ester. The preparation and ex vivo evaluation of this new beta-CIT-analog ([18F]FE@CIT) is presented here. Precursor and standard were prepared from beta-CIT and analyzed by spectroscopic methods. Yields of precursor and standard preparation were 61% and 42%, respectively. [18F]FE@CIT was prepared by distillation of [18F]bromofluoroethane ([18F]BFE) and reaction with (1R-2-exo-3-exo)8-methyl-3-(4 iodo-phenyl)-8-azabicyclo[3.2.1] octane-2-carboxylic acid. After 10 min at 150 degrees C the product was purified using a C-18 SepPak. The radiosynthesis evinced radiochemical yields of >90% (based on [18F]BFE), the specific radioactivity was >416 GBq/micromol. An average 30 microAh cyclotron irradiation yielded more than 2.5 GBq [18F]FE@CIT. For the ex vivo bioevaluation, 20 male Sprague-Dawley rats were sacrificed at 5, 15, 30, 60, and 120 min after injection. Organs were removed, weighed, and counted. For autoradiographic experiments, transverse brain slices of about 100 microm were prepared. The ex vivo evaluation showed highest brain uptake in striatal regions, followed by thalamus and cerebellum. The highest striatum to cerebellum ratio was 3.73 and the highest thalamus to cerebellum ratio was 1.65. Autoradiographic images showed a good and differentiated uptake in striatal regions with a good target-to background ratio. PMID- 15529337 TI - Utilization of a new biotinylation reagent in the development of a nondiscriminatory investigative approach for the study of cell surface proteins. AB - In order to circumvent the various problems encountered during the study of membrane-bound proteins, we designed and synthesized a novel membrane-impermeable biotinylation reagent incorporating chemical properties compatible with this goal. We then developed a nondiscriminatory analytical procedure for such studies which overcomes possible selectivity, contamination and solubility problems. The necessary steps (labeling, limited in situ proteolysis, affinity purification) are all conducted in mild or near native conditions. This versatile method could provide an accurate picture of the cell surface proteome. PMID- 15529338 TI - Global proteomic approach unmasks involvement of keratins 8 and 18 in the delivery of cystic fibrosis transmembrane conductance regulator (CFTR)/deltaF508 CFTR to the plasma membrane. AB - Cystic fibrosis (CF) is a genetic disease caused by mutations in the CF gene (cftr). Physiologically, CF is characterized by an abnormal chloride secretion in epithelia due to a dysfunction of a mutated cystic fibrosis transmembrane conductance regulator (CFTR). CFTR is a cAMP-dependent chloride channel whose most frequent mutation, deltaF508, leads to an aberrantly folded protein which causes a dysfunction of the channel. However, a growing number of reports suggest that modifier genes and environmental factors are involved in the physiology of CF. To identify proteins whose expression depends on wild-type WT-CFTR or deltaF508-CFTR, we chose a global proteomic approach based on the use of two dimensional gel electrophoresis (2-DE) and mass spectrometry. The experiments were carried out with HeLa cells stably transfected with WT-CFTR (pTCFWT) or deltaF508-CFTR (pTCFdeltaF508). These experiments unmasked keratin 8 (K8) and 18 (K18) which were differentially expressed in pTCFWT vs. pTCFdeltaF508. An immunoblot of K18 confirmed the 2-DE results. Intracellular localization experiments of WT-CFTR, deltaF508-CFTR, K8, and K18 suggest that the expression of these proteins are linked, and that the concentrations of K8 and K18 and/or their distribution may be involved in the traffic of WT-CFTR/deltaF508-CFTR. A functional assay for CFTR revealed that specifically lowering K18 expression or changing its distribution leads to the delivery of functional deltaF508-CFTR to the plasma membrane. This work suggests a novel function of K18 in CF. PMID- 15529339 TI - Clinical images: Bone marrow edema syndrome. PMID- 15529341 TI - The synovium in rheumatoid arthritis: comment on the editorial by Kirwan. PMID- 15529342 TI - Anakinra in mutation-negative NOMID/CINCA syndrome: comment on the articles by Hawkins et al and Hoffman and Patel. PMID- 15529343 TI - Minocycline-induced hyperpigmentation masquerading as alkaptonuria in individuals with joint pain. AB - Alkaptonuria, a rare autosomal-recessive disorder caused by mutations in the HGD gene and a deficiency of homogentisate 1,2-dioxygenase, is characterized by accumulation of homogentisic acid (HGA), ochronosis, and destruction of connective tissue resulting in joint disease. Certain medications have been reported to cause cutaneous hyperpigmentation resembling that of alkaptonuria. We present 5 such cases. Eighty-eight patients with a possible diagnosis of alkaptonuria were examined at the National Institutes of Health Clinical Center between June 2000 and March 2004. The diagnosis of alkaptonuria was confirmed or ruled out by measurement of HGA in the urine. Five patients with findings consistent with ochronosis, including pigmentary changes of the ear and mild degenerative disease of the spine and large joints, were diagnosed clinically as having alkaptonuria, but the diagnosis was withdrawn based on normal urine HGA levels. All 5 patients were women who had taken minocycline for dermatologic or rheumatologic disorders for extended periods. Minocycline-induced hyperpigmentation should be considered in the differential diagnosis of ochronosis. This could be of increased significance now that minocycline and other tetracyclines have been proposed as therapeutic options for rheumatoid arthritis, bringing a new population of patients with ochronosis and arthritis to medical attention with the potential, but incorrect, diagnosis of alkaptonuria. PMID- 15529344 TI - Identification of new quantitative trait loci in mice with collagen-induced arthritis. AB - OBJECTIVE: Collagen-induced arthritis (CIA) in the mouse is one of the most widely used autoimmune experimental models, with many features similar to rheumatoid arthritis. This study sought to identify potential genetic regulatory mechanisms of CIA in major histocompatibility complex-matched (H2-q) F(2) hybrid mice. METHODS: We used 126 polymorphic markers to perform simple sequence-length polymorphism analysis on 290 F(2) hybrids of arthritis-susceptible (DBA/1J) and arthritis-resistant (FVB/N) inbred mouse strains. The major clinical traits (disease severity and onset) were assessed, and serum antibodies specific to type II collagen (CII) were determined by enzyme-linked immunosorbent assay in 270 F(2) mice. Lymph nodes from 94 F(2) mice were used to test the ratio of CD4 to CD8 by fluorescence-activated cell sorter analysis, and cell proliferation was determined by XTT test. RESULTS: Two quantitative trait loci (QTLs) identified in previous studies were confirmed; these were severity-controlling Cia2 and onset controlling Cia4 on chromosome 2. Moreover, we identified 5 new QTLs, 1 for CII specific IgG2a antibodies on chromosome 5, 2 controlling the CII-specific IgG1 antibody response on chromosomes 10 and 13, 1 for the CD4:CD8 ratio on chromosome 2, and 1 for cell proliferation (measured by XTT test) on chromosome 16. Complement component C5 was identified as the probable main candidate gene for the QTLs Cia2 and Cia4. F(2) mice carrying a 2-basepair deletion of C5, the FVB/N allele, had low incidence and less severe disease as compared with those carrying the DBA/1J allele. CONCLUSION: This genome scan provides additional evidence confirming the role of C5 as a probable candidate gene for Cia2 and Cia4 loci, and identifies new QTLs controlling new traits in autoimmune arthritis. PMID- 15529345 TI - The combination of SOX5, SOX6, and SOX9 (the SOX trio) provides signals sufficient for induction of permanent cartilage. AB - OBJECTIVE: To regenerate permanent cartilage, it is crucial to know not only the necessary conditions for chondrogenesis, but also the sufficient conditions. The objective of this study was to determine the signal sufficient for chondrogenesis. METHODS: Embryonic stem cells that had been engineered to fluoresce upon chondrocyte differentiation were treated with combinations of factors necessary for chondrogenesis, and chondrocyte differentiation was detected as fluorescence. We screened for the combination that could induce fluorescence within 3 days. Then, primary mesenchymal stem cells, nonchondrogenic immortalized cell lines, and primary dermal fibroblasts were treated with the combination, and the induction of chondrocyte differentiation was assessed by detecting the expression of the cartilage marker genes and the accumulation of proteoglycan-rich matrix. The effects of monolayer, spheroid, and 3-dimensional culture systems on induction by combinations of transcription factors were compared. The effects of the combination on hypertrophic and osteoblastic differentiation were evaluated by detecting the expression of the characteristic marker genes. RESULTS: No single factor induced fluorescence. Among various combinations examined, only the SOX5, SOX6, and SOX9 combination (the SOX trio) induced fluorescence within 3 days. The SOX trio successfully induced chondrocyte differentiation in all cell types tested, including nonchondrogenic types, and the induction occurred regardless of the culture system used. Contrary to the conventional chondrogenic techniques, the SOX trio suppressed hypertrophic and osteogenic differentiation at the same time. CONCLUSION: These data strongly suggest that the SOX trio provides signals sufficient for the induction of permanent cartilage. PMID- 15529346 TI - Rituximab improves peripheral B cell abnormalities in human systemic lupus erythematosus. AB - OBJECTIVE: B lymphocyte depletion has recently emerged as a promising approach to the treatment of systemic lupus erythematosus (SLE). As part of a phase I/II dose ranging trial of rituximab in the treatment of SLE, we evaluated the fate of discrete B cell subsets in the setting of selective depletion by anti-CD20 monoclonal antibody and during the B cell recovery phase. METHODS: B cell depletion and phenotype were examined by flow cytometry of peripheral blood mononuclear cells for CD19, CD20, CD27, IgD, and CD38 expression. Changes in autoreactive B lymphocytes and plasma cells were assessed by determination of serum autoantibody levels (anti-double-stranded DNA and VH4.34) and by direct monitoring of a unique autoreactive B cell population bearing surface antibodies whose heavy chain is encoded by the VH4.34 gene segment. RESULTS: Compared with normal controls, SLE patients displayed several abnormalities in peripheral B cell homeostasis at baseline, including naive lymphopenia, expansion of a CD27 ,IgD- (double negative) population, and expansion of circulating plasmablasts. Remarkably, these abnormalities resolved after effective B cell depletion with rituximab and immune reconstitution. The frequency of autoreactive VH4.34 memory B cells also decreased 1 year posttreatment, despite the presence of low levels of residual memory B cells at the point of maximal B cell depletion and persistently elevated serum autoantibody titers in most patients. CONCLUSION: This study is the first to show evidence that in SLE, specific B cell depletion therapy with rituximab dramatically improves abnormalities in B cell homeostasis and tolerance that are characteristic of this disease. The persistence of elevated autoantibody titers may reflect the presence of low levels of residual autoreactive memory B cells and/or long-lived autoreactive plasma cells. PMID- 15529348 TI - Bone morphogenetic protein and transforming growth factor beta inhibitory Smads 6 and 7 are expressed in human adult normal and osteoarthritic cartilage in vivo and are differentially regulated in vitro by interleukin-1beta. AB - OBJECTIVE: Bone morphogenetic protein (BMP) and transforming growth factor beta (TGFbeta) are potent anabolic factors in adult articular chondrocytes. In this study, we investigated whether intracellular inhibitors of BMP and TGFbeta signaling, inhibitory Smad6 (I-Smad6) and I-Smad7, are expressed in articular chondrocytes in normal and osteoarthritic (OA) cartilage, and whether their expression shows a correlation with the anabolic activity of OA chondrocytes in vivo and after interleukin-1beta (IL-1beta) stimulation in vitro. METHODS: RNA isolated directly from normal and OA human knee cartilage as well as from cultured articular chondrocytes was analyzed by (quantitative) polymerase chain reaction technology. Immunolocalization of the I-Smads was performed on tissue sections and compared with the anabolic cellular activity as documented by in situ hybridization experiments for aggrecan and type II collagen. RESULTS: Both Smad6 and Smad7 were expressed in all samples of normal and OA cartilage. Immunostaining (including confocal microscopy) confirmed the presence of Smad6 and Smad7 in the majority of normal and degenerated articular chondrocytes; localization was mostly cytoplasmic. No correlation between expression of the main anabolic genes and expression of the I-Smads was found. In cultured articular chondrocytes, stimulation with IL-1beta showed up-regulation of Smad7, whereas Smad6 was down-regulated. CONCLUSION: Both Smad6 and Smad7 are expressed in adult human articular chondrocytes. The primarily cytoplasmic localization suggests permanent activation of the I-Smads in articular cartilage in vivo. No evidence was found that up-regulation or down-regulation of I-Smads in OA cartilage correlates directly with the anabolic (or catabolic) activity of articular chondrocytes. The regulation in chondrocytes of Smad6 and Smad7 expression by IL-1beta suggests a potentially important role of IL-1beta signaling in chondrocytes, via indirect influencing of the BMP/TGFbeta signaling cascade. PMID- 15529349 TI - Autoreactive CD8+ cytotoxic T lymphocytes to major histocompatibility complex class I chain-related gene A in patients with Behcet's disease. AB - OBJECTIVE: To detect and characterize the autoreactive CD8+ T cells to major histocompatibility complex class I chain-related gene A (MICA), a stress inducible antigen preferentially expressed on the epithelium and endothelium, in patients with Behcet's disease (BD). METHODS: A candidate for the antigenic MICA peptide was selected based on its predicted binding affinity for HLA-B51 and proteasomal cleavage sites. Peripheral blood T cells from 14 patients with BD and 15 healthy controls were repeatedly stimulated with the MICA peptide, and the specific T cell response was measured by peptide-induced interferon-gamma. Cytotoxic T lymphocyte activity was examined by chromium-51 release from an HLA B51-transfected B cell line in the presence of the MICA peptide. RESULTS: A 9-mer peptide AAAAAIFVI (termed MICA transmembrane [MICA-TM]) was selected as a candidate for the antigenic peptide presented by HLA-B51. A specific T cell response to MICA-TM was detected in 4 patients with BD (29%) but in none of the 15 healthy donors. All 4 responders had HLA-B51 and active disease, and the specific T cell response was lost after the BD-related symptoms disappeared. The MICA-induced T cell response was specifically inhibited by anti-HLA class I antibody or by CD8+ cell depletion. MICA-reactive T cells recognized an HLA-B51 transfected B cell line pulsed with MICA-TM or a B cell line transfected with both HLA-B51 and MICA in the absence of exogenous peptides. Finally, MICA stimulated T cell lines lysed the HLA-B51-expressing B cell line in the presence of MICA-TM. CONCLUSION: HLA-B51-restricted cytotoxic T lymphocytes autoreactive to MICA may be involved in the pathogenesis of BD. PMID- 15529350 TI - Relative ineffectiveness of triamcinolone acetonide in the treatment of juvenile idiopathic arthritis. PMID- 15529351 TI - Do breast-feeding and other reproductive factors influence future risk of rheumatoid arthritis? Results from the Nurses' Health Study. AB - OBJECTIVE: To explore the contribution of female hormonal factors occurring prior to the onset of rheumatoid arthritis (RA), such as age at menarche, parity, age at first birth, breast-feeding, use of oral contraceptives (OCs), irregular menstrual cycles, and postmenopausal hormone (PMH) use, to the subsequent development of RA in a large female cohort. METHODS: We studied female reproductive and hormonal risk factors for RA in a cohort of 121,700 women enrolled in the longitudinal Nurses' Health Study. The diagnosis of incident RA (between 1976 and 2002) in 674 women was confirmed by a connective tissue disease screening questionnaire and blinded medical record review for American College of Rheumatology criteria. Sixty percent of the patients with RA were rheumatoid factor positive. The relationship between potential risk factors, including age, age at menarche, parity, age at first birth, total lifetime history of breast feeding, use of OCs, and irregular menstrual cycles and the multivariate-adjusted risk of RA was estimated using Cox proportional hazards models. RESULTS: Using a multivariate model that adjusted for age, body mass index, smoking, parity, and other hormonal factors, we observed a strong trend for decreasing risk of RA with increasing duration of breast-feeding (P for trend = 0.001). For women who breast fed (compared with parous women who did not breast-feed), the risk ratios (RRs) and 95% confidence intervals (95% CIs) were as follows: breast-feeding for < or =3 total months, RR 1.0 (95% confidence interval [95% CI] 0.8-1.2); for 4-11 total months, RR 0.9 (95% CI 0.7-1.1); for 12-23 total months, RR 0.8 (95% CI 0.6 1.0); and for > or =24 total months, RR 0.5 (95% CI 0.3-0.8). Very irregular menstrual cycles were associated with an increased risk of RA (RR 1.4, 95% CI 1.0 2.0). Age at menarche < or =10 years was associated with an increased risk of seropositive RA (RR 1.6, 95% CI 1.1-2.4) but not significantly associated with risk of RA. Parity, total number of children, age at first birth, and OC use were not associated with an increased risk of RA in this cohort. CONCLUSION: In this large cohort, breast-feeding for >12 months was inversely related to the development of RA. This apparent effect was dose-dependent, with a significant trend toward lower risk with longer duration of breast-feeding. Irregular menstrual cycles and earlier age at menarche increased the risk of RA. Other reproductive hormonal factors were not associated with RA risk. PMID- 15529352 TI - Particular HLA-DRB1 shared epitope genotypes are strongly associated with rheumatoid vasculitis. AB - OBJECTIVE: To examine the relationship of the HLA-DRB1 shared epitope (SE) to rheumatoid vasculitis, using individual patient data (IPD) meta-analytic methods. METHODS: Published studies that enrolled adult patients with rheumatoid arthritis (RA) were identified by searches of Medline and Embase, and by manual searches of medical journals. All authors were contacted for IPD. Meta-analyses were performed to assess the association of SE presence, dose, and genotype with rheumatoid vasculitis. RESULTS: A total of 14 studies and 1,568 patients (129 with vasculitis) were included in the analysis. RA patients with vasculitis were significantly more likely to have rheumatoid nodules (odds ratio [OR] 2.5, 95% confidence interval [95% CI] 1.5-3.9], but there was no significant association with male sex, rheumatoid factor positivity, or erosive disease. No significant association was observed between the presence of the SE (i.e., 1 or 2 alleles versus 0 alleles) and rheumatoid vasculitis (summary OR 1.4, 95% CI 0.7-2.7). Analysis by SE genotype, however, demonstrated a striking relationship of vasculitis to 3 genotypes containing a double dose of the SE, specifically HLA DRB1*0401/*0401 (OR 6.2, 95% CI 1.01-37.9), *0401/*0404 (OR 4.1, 95% CI 1.1 16.2), and *0101/*0401 (OR 4.0, 95% CI 1.4-11.6). CONCLUSION: The HLA-DRB1 SE genotypes *0401/*0401, *0401/*0404, and *0101/*0401 may be of particular importance to rheumatoid vasculitis. It is hoped that additional investigation of these and other SE genotypes will lead to improved insight into the mechanisms influencing the clinical expression of RA. PMID- 15529353 TI - Antiarthritic effect of bee venom: inhibition of inflammation mediator generation by suppression of NF-kappaB through interaction with the p50 subunit. AB - OBJECTIVE: To investigate the molecular mechanisms of the antiarthritic effects of bee venom (BV) and melittin (a major component of BV) in a murine macrophage cell line (Raw 264.7) and in synoviocytes obtained from patients with rheumatoid arthritis. METHODS: We evaluated the antiarthritic effects of BV in a rat model of carrageenan-induced acute edema in the paw and in a rat model of chronic adjuvant-induced arthritis. The inhibitory effects of BV and melittin on inflammatory gene expression were measured by Western blotting, and the generation of prostaglandin E(2) (PGE(2)) and nitric oxide (NO) and the intracellular calcium level were assayed. NF-kappaB DNA binding and transcriptional activity were determined by gel mobility shift assay or by luciferase assay. Direct binding of BV and melittin to the p50 subunit of NF kappaB was determined with a surface plasmon resonance analyzer. RESULTS: BV (0.8 and 1.6 mug/kg) reduced the effects of carrageenan- and adjuvant-induced arthritis. This reducing effect was consistent with the inhibitory effects of BV (0.5, 1, and 5 mug/ml) and melittin (5 and 10 mug/ml) on lipopolysaccharide (LPS; 1 mug/ml)-induced expression of cyclooxygenase 2, cytosolic phospholipase A(2), inducible NO synthase, generation of PGE(2) and NO, and the intracellular calcium level. BV and melittin prevented LPS-induced transcriptional and DNA binding activity of NF-kappaB via the inhibition of IkappaB release and p50 translocation. BV (affinity [K(d)] = 4.6 x 10(-6)M) and melittin (K(d) = 1.2 x 10(-8)M) bound directly to p50. CONCLUSION: Target inactivation of NF-kappaB by directly binding to the p50 subunit is an important mechanism of the antiarthritic effects of BV. PMID- 15529355 TI - Smoking and increased apoptosis in patients with systemic lupus erythematosus: comment on the article by Costenbader et al. PMID- 15529357 TI - No decrease over time in the incidence of vasculitis or other extraarticular manifestations in rheumatoid arthritis: results from a community-based study. PMID- 15529356 TI - Lipopolysaccharide-induced expression of multiple alternatively spliced MEFV transcripts in human synovial fibroblasts: a prominent splice isoform lacks the C terminal domain that is highly mutated in familial Mediterranean fever. AB - OBJECTIVE: To investigate the expression of the familial Mediterranean fever (FMF) gene (MEFV) in human synovial fibroblasts. METHODS: MEFV messenger RNA in synovial fibroblasts, chondrocytes, and peripheral blood leukocytes (PBLs) was analyzed by semiquantitative and real-time polymerase chain reaction and ribonuclease protection assay. The subcellular localization of pyrin, the MEFV product, was determined in transfected synovial fibroblasts and HeLa cells with plasmids encoding pyrin isoforms. Native pyrin was detected with an antipyrin antibody. RESULTS: MEFV was expressed in synovial fibroblasts, but not in chondrocytes. Four alternatively spliced transcripts were identified: an extension of exon 8 (exon 8ext) resulting in a frameshift that predicts a truncated protein lacking exons 9 and 10, the addition of an exon (exon 4a) predicting a truncated protein at exon 5, the in-frame substitution of exon 2a for exon 2, and the previously described removal of exon 2 (exon 2Delta). Exon 8ext transcripts represented 27% of the total message population in synovial fibroblasts. All other alternatively spliced transcripts were rare. Consensus and alternatively spliced transcripts were induced by lipopolysaccharide in synovial fibroblasts and PBLs. In transfected cells, the proteins encoded by all highly expressed splice forms were cytoplasmic. In contrast, native pyrin was predominantly nuclear in synovial fibroblasts, neutrophils, and dendritic cells, but was cytoplasmic in monocytes. CONCLUSION: Several MEFV transcripts are expressed and inducible in synovial fibroblasts. A prominent isoform lacks the C terminal domain that contains the majority of mutations found in patients with FMF. While recombinant forms of all major pyrin isoforms are cytoplasmic, native pyrin is nuclear in several cell types. Thus, mechanisms in addition to splicing patterns must control pyrin's subcellular distribution. PMID- 15529358 TI - Interferon-inducible protein 10/CXCL10 is increased in the cerebrospinal fluid of patients with central nervous system lupus. PMID- 15529359 TI - Clonal B cell populations in the blood and liver of patients with chronic hepatitis C virus infection. AB - OBJECTIVE: The association of hepatitis C virus (HCV) infection with type II mixed cryoglobulinemia is well established, but the role of HCV in B cell lymphoma remains controversial. The objective of this study was to determine the frequency of circulating and liver-infiltrating monoclonal B cells in patients with HCV infection. METHODS: One hundred sixty patients were studied prospectively, including 115 HCV-positive patients and 45 HCV-negative patients with other nonimmune chronic liver disease(s). B cell clonality was determined by DNA amplification of the IgH rearrangements, followed by polyacrylamide gel electrophoresis. RESULTS: A clonal B cell population was detected in the blood of 21 (26%) of 81 HCV-positive patients whose cryoglobulin status was known, including 12 of 25 patients with type II cryoglobulinemia, 2 of 12 patients with type III cryoglobulinemia, and 7 of 44 patients without cryoglobulins. A clonal IgH rearrangement was detected in 26 (32%) of 81 liver biopsy specimens from HCV- infected patients, including 16 patients with a circulating clonal population. A clonal B cell population was not observed in the blood of 40 patients with non HCV liver diseases and was present in only 1 (3%) of 30 liver biopsy specimens. Logistic regression analysis showed that HCV-infected patients with clonal B cell proliferation in both the blood and liver were older (P = 0.004) and had longer duration of HCV infection (P = 0.009), higher serum cryoglobulin levels (P = 0.001) that were more frequently symptomatic (P < 0.03), and liver disease that was more severe than that in patients without a clonal B cell population in the blood or liver (P = 0.05). In 4 of 16 patients with a clonal B cell population in both the blood and liver, a definite B cell malignancy was finally diagnosed. CONCLUSION: Clonal B lymphocytes are frequently detected in the blood and liver of patients with chronic HCV infection, in the absence of overt B cell malignancy. These clones are usually, but not always, associated with the presence of type II cryoglobulins. A high percentage of patients with B cell clonality in both the blood and liver were finally diagnosed as having a definite B cell malignancy. PMID- 15529360 TI - Inhibition of adjuvant-induced arthritis by DNA vaccination with the 70-kd or the 90-kd human heat-shock protein: immune cross-regulation with the 60-kd heat-shock protein. AB - OBJECTIVE: Adjuvant arthritis can be induced in Lewis rats by immunization with Mycobacterium tuberculosis (Mt). The mycobacterial 65-kd heat-shock protein (Hsp65) is targeted by arthritogenic T cells. However, Hsp65 and the mycobacterial 71-kd heat-shock protein are also recognized by T cells that can down-regulate adjuvant-induced arthritis (AIA). We have recently demonstrated that vaccination with human Hsp60 DNA inhibits AIA. The present study was undertaken to analyze the role of the T cell responses to self HSP molecules other than Hsp60 in the control of AIA. METHODS: Lewis rats were immunized with DNA vaccines coding for human Hsp70 or Hsp90 (Hsp70 plasmid [pHsp70] or pHsp90), and AIA was induced. The T cell response to Mt, Hsp60, Hsp70, and Hsp90 (proliferation and cytokine release) was studied, and the T cell response to Hsp60 was mapped with overlapping peptides. RESULTS: The Hsp70 or Hsp90 DNA vaccines shifted the arthritogenic T cell response from a Th1 to a Th2/3 phenotype and inhibited AIA. We detected immune crosstalk between Hsp70/90 and Hsp60: both the Hsp70 and Hsp90 DNA vaccines induced Hsp60-specific T cell responses. Similarly, DNA vaccination with Hsp60 induced Hsp70-specific T cell immunity. Epitope mapping studies revealed that Hsp60-specific T cells induced by pHsp70 vaccination reacted with known regulatory Hsp60 epitopes. CONCLUSION: T cell immunity to Hsp70 and to Hsp90, like Hsp60-specific immunity, can modulate the arthritogenic response in AIA. In addition, our results suggest that the regulatory mechanisms induced by Hsp60, Hsp70, and Hsp90 are reinforced by an immune network that connects their reactivities. PMID- 15529361 TI - Immunogenetic differences between Caucasian women with and those without silicone implants in whom myositis develops. AB - OBJECTIVE: To determine whether patients in whom myositis develops after they receive silicone breast implants have distinct clinical, serologic, and/or immunogenetic features compared with patients with myositis who do not have silicone implants. METHODS: A preliminary case series study was followed by a larger, independent, matched case-control study to evaluate women in whom myositis developed after they received silicone implants (MASI patients) compared with healthy women with silicone implants and women with myositis but without silicone implants (idiopathic inflammatory myopathy; IIM patients). RESULTS: In a preliminary study, 11 MASI patients differed from 76 IIM patients in having an increased frequency of HLA-DQA1*0102 (odds ratio [OR] 9.8, 95% confidence interval [95% CI] 1.77-96.79) and decreased frequencies of the myositis associated risk factor DRB1*0301 (OR 0.1 [95% CI 0.002-0.63]) and its linked allele DQA1*0501 (OR 0.2 [95% CI 0.02-0.87]). A subsequent independent, matched case-control study revealed that although clinical features and autoantibodies did not differ significantly between the MASI and IIM groups, MASI patients again had decreased frequencies of DRB1*0301 (OR 0.2 [95% CI 0.07-0.72]) and DQA1*0501 (OR 0.2 [95% CI 0.08-0.84]) compared with IIM patients. Additional comparisons between MASI patients from both studies combined (n = 37) and a larger population of IIM patients (n = 453) suggested that HLA-DQA1*0102 may be uniquely associated with MASI (OR 2.6 [95% CI 1.25-5.46]). CONCLUSION: Women in whom inflammatory myopathy develops after they receive silicone implants constitute an immunogenetically distinct group of patients with myositis. These and other data suggest that autoimmune diseases as now defined may consist of multiple distinct entities, each of which is characterized by different genes and environmental exposures. PMID- 15529362 TI - Serum cytokine profiles in relapsing polychondritis suggest monocyte/macrophage activation. AB - OBJECTIVE: There is evidence that autoimmunity plays a significant role in the pathogenesis of relapsing polychondritis (RP). This study was designed to investigate circulating levels of various cytokines in relation to the etiology of this rare disorder, and to compare the pattern of cytokine elevations in RP with that in another autoimmune disease, rheumatoid arthritis (RA). METHODS: Serum from 22 patients with active RP and an equal number of age- and sex-matched healthy controls and RA patients were available for analysis. The following cytokines were measured: interleukin-1beta (IL-1beta), IL-2, IL-4, IL-5, IL-6, IL 7, IL-8, IL-10, IL-12, IL-13, IL-17, interferon-gamma (IFNgamma), tumor necrosis factor alpha, granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), monocyte chemoattractant protein 1 (MCP-1), and macrophage inflammatory protein 1beta (MIP-1beta). Results were analyzed by nonparametric Mann-Whitney test with Holm stepdown adjustment for multiple testing. RESULTS: The levels of 3 of these cytokines showed significant differences between RP patients and controls. Compared with controls, mean serum levels of MCP-1, MIP-1beta, and IL-8 were all much higher in patients with active RP. In contrast, RA patients showed a more general increase in all cytokines measured, with much higher levels of IL-2, IL-4, IL-5, IL-6, IL-7, IL-10, IL-13, IFNgamma, G-CSF, GM-CSF, MCP-1, and MIP-1beta compared with controls. CONCLUSION: Levels of 3 serum cytokines were significantly higher in RP patients than in age- and sex-matched controls. One of these 3 cytokines, IL-8, was not significantly elevated in RA samples. Overall, in RP, a more discrete group of cytokines exhibited significantly increased levels than was found in RA. Each of the 3 cytokines that were elevated in RP is a proinflammatory chemokine, characteristic of activation of the monocyte and macrophage lineage, and in the case of IL-8, also of neutrophils. These data suggest a major role for a cell-mediated immune response in the pathophysiology of RP. PMID- 15529363 TI - Increased susceptibility to rheumatoid arthritis in Koreans heterozygous for HLA DRB1*0405 and *0901. AB - OBJECTIVE: To investigate the association of susceptibility and protective HLA DRB1 alleles with rheumatoid arthritis (RA) and its clinical markers in an Asian population. METHODS: All RA patients (n = 574) and control subjects (n = 392) were Korean. HLA-DRB1 typing and further subtyping of all alleles was performed by polymerase chain reaction, sequence-specific oligonucleotide probe hybridization, and direct DNA sequencing analysis. We used a relative predispositional effects (RPEs) method and a false discovery rate correction method for multiple comparisons. RESULTS: The DRB1*0405 and *0901 alleles showed the most significant associations with RA (P = 7.83 x 10(-24), odds ratio [OR] 4.40 [95% confidence interval (95% CI) 3.24-5.99], and P = 3.76 x 10(-9), OR 2.47 [95% CI 1.82-3.36], respectively). The RPEs test showed that the DRB1*0401 and *0410 alleles conferred susceptibility and that the DRB1*0701, *0802, *1301, *1302, *1403, and *1405 alleles showed significant protective effects. Susceptibility and protective alleles both showed a pattern consistent with additive genetic effects, and each influenced RA independently of the other. The compound heterozygote DRB1*0405/*0901 was associated with the highest risk of RA (corrected P = 1.81 x 10(-11), OR 58.2 [95% CI 7.95-425.70]). The mean age at disease onset was approximately 4 years earlier or was 3 years earlier, respectively, in patients with at least 1 copy of the DRB1*0405 or the DRB1*0901 allele. Radiographic changes (stages II-IV) were more frequent in patients with at least 1 copy of DRB1*0405 (P = 0.032, 92.6% versus 84.3%, OR 2.33 [95% CI 1.24 4.39]). CONCLUSION: The DRB1*0405/*0901 heterozygote has the strongest association with RA, suggesting that this heterozygote enhances the susceptibility to RA in Koreans. PMID- 15529364 TI - Measurement of erythrocyte C4d and complement receptor 1 in systemic lupus erythematosus. AB - OBJECTIVE: C4-derived activation fragments are the only complement ligands present on the surfaces of normal erythrocytes. The significance of this observation is unknown, and the role of erythrocyte-bound C4 (E-C4) in human disease has not been explored. More than any other human disease, the pathogenesis of systemic lupus erythematosus (SLE) has been characterized by defects in clearance of complement-bearing immune complexes via erythrocytes expressing complement receptor 1 (CR1). This study was undertaken to determine whether these functional defects might be reflected by abnormal patterns of E-C4 and E-CR1 expression on erythrocytes of patients with SLE. METHODS: We conducted a cross-sectional study of 100 patients with SLE, 133 patients with other diseases, and 84 healthy controls. Erythrocytes were characterized by indirect immunofluorescence and by flow cytometry for determination of levels of C4d and CR1. RESULTS: Patients with SLE had higher levels of E-C4d and lower levels of E CR1 than did patients with other diseases (P < or = 0.001) or healthy controls (P < or = 0.001). The test was 81% sensitive and 91% specific for SLE versus healthy controls and 72% sensitive and 79% specific for SLE versus other diseases, and it had an overall negative predictive value of 92%. CONCLUSION: This is the first report of abnormal levels of E-C4d in human disease. We found that abnormally high levels of E-C4d and low levels of E-CR1 are characteristic of SLE, and combined measurement of the 2 molecules has high diagnostic sensitivity and specificity for lupus. Determination of E-C4d/E-CR1 levels may be a useful addition to current tests and criteria for SLE diagnosis. PMID- 15529365 TI - Antimicrobial proteins act as "alarmins" in joint immune defense. PMID- 15529366 TI - Glucocorticoids in the treatment of rheumatic diseases: an update on the mechanisms of action. PMID- 15529367 TI - The relationship of antiresorptive drug use to structural findings and symptoms of knee osteoarthritis. AB - OBJECTIVE: To examine the cross-sectional association between use of medications that have a bone antiresorptive effect (estrogen, raloxifene, and alendronate) and both the structural features of knee osteoarthritis (OA), assessed by magnetic resonance imaging (MRI) and radiography, and the symptoms of knee OA in elderly women. METHODS: Women in the Health, Aging and Body Composition Study underwent MRI and radiography of the knee if they reported symptoms of knee OA, and women without significant knee symptoms were selected as controls. MR images of the knee were assessed for multiple features of OA using the Whole-Organ MRI scoring method, and radiographs were read for Kellgren and Lawrence grade and individual features of OA. Concurrent medication use and knee symptoms were assessed by interview, and knee pain severity was evaluated using the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). RESULTS: There were 818 postmenopausal women from whom we obtained MR images of the knee and data on medication use. Among these women, 214 (26.2%) were receiving antiresorptive drugs. We found no significant association between overall use of antiresorptive drugs and the presence of knee pain and radiographic changes of OA of the knee. Use of alendronate, but not estrogen, was associated with less severity of knee pain as assessed by WOMAC scores. Both alendronate use and estrogen use were associated with significantly less subchondral bone attrition and bone marrow edema-like abnormalities in the knee as assessed by MRI, as compared with women who had not received these medications. CONCLUSION: Elderly women being treated with alendronate and estrogen had a significantly decreased prevalence of knee OA-related subchondral bone lesions compared with those reporting no use of these medications. Alendronate use was also associated with a reduction in knee pain according to the WOMAC scores. PMID- 15529368 TI - An alternate hypothesis regarding radiologic damage to synovial tissue: comment on the editorial by Kirwan. PMID- 15529369 TI - Association between nuclear antigens and endogenous retrovirus in the generation of autoantibody responses in murine lupus. AB - OBJECTIVE: (NZB x NZW)F(1) (NZB/NZW) mice and other strains of mice with experimental lupus frequently produce autoantibodies to both chromatin constituents and murine leukemia virus envelope gp70. These autoantibody responses are involved in the glomerulonephritis that develops in these mice. This study was undertaken to explore possible connections between these 2 antigen systems. METHODS: We used monoclonal antibodies (mAb) derived from unmanipulated NZB/NZW mice to investigate the specificity of anti-gp70 and antichromatin autoantibodies for chromatin constituents, recombinant gp70, NZB retroviruses, and retrovirally infected cells. NZB mice were also immunized with retroviral particles and followed up for study of autoantibody responses. RESULTS: Spontaneous autoantibody production in NZB/NZW mice reflects high-level autoimmune responses to nuclear antigens and gp70 that do not cross-react with the other antigen. However, both types of autoantibodies have the capability to bind to the endogenous xenotropic virions NZB-X1 or NZB-X2. The mAbs to recombinant gp70 cross-reacted only with the NZB-X2 virus, whereas the antichromatin mAb frequently bound to both retroviruses. The binding of antichromatin autoantibodies was mediated by nuclear material complexed to the retrovirus, and studies showed that this material can be acquired through the budding process. Immunization with NZB-X1 or NZB-X2 virions induced strong responses to gp70 and was much more effective than chromatin at inducing autoantibody responses to chromatin and double-stranded DNA in NZB mice. CONCLUSION: These studies suggest that retroviral virions may harbor nuclear antigens and may link together the autoimmune responses to the disparate antigens, chromatin and gp70. PMID- 15529370 TI - Role of alendronate in therapy for posttraumatic complex regional pain syndrome type I of the lower extremity. AB - OBJECTIVE: To evaluate the effects of the antiresorptive agent alendronate at a daily oral dose of 40 mg in patients with posttraumatic complex regional pain syndrome type I (CRPS I) of the lower extremity. METHODS: Forty patients were enrolled in this 8-week randomized, double-blind, placebo-controlled study of alendronate therapy for CRPS I, a condition associated with regional osteoclastic overactivity. An optional 8-week open extension of alendronate therapy (weeks 12 20) was available after a 4-week period without therapy. Clinical assessments included joint mobility, edema of the lower extremity, tolerance to pressure in the lower extremity, and levels of spontaneous pain. Urinary levels of type I collagen N-telopeptide (NTX) were assessed by enzyme-linked immunosorbent assay. Patients were examined at weeks 4, 8, 12, 16, 20, and 24. Statistical analysis included two-way factorial analysis of variance. RESULTS: In contrast to placebo treated patients (n = 20), all of the alendronate-treated patients (n = 19) exhibited a marked and sustained improvement in levels of spontaneous pain, pressure tolerance, and joint mobility, as well as a significant reduction in urinary levels of NTX at weeks 4 and 8. The improvement was maintained at week 12. Twelve patients from each treatment group volunteered for the 8-week open trial, and all of them had a positive response to alendronate. CONCLUSION: Our findings support the use of oral alendronate in posttraumatic CRPS I. By reducing local acceleration of bone remodeling, alendronate might relieve pain by effects on nociceptive primary afferents in bone, pain-associated changes in the spinal cord, and possibly also through a central mechanism. PMID- 15529371 TI - Anti-lipoprotein lipase antibodies: a new player in the complex atherosclerotic process in systemic lupus erythematosus? AB - OBJECTIVE: The novel description of antibodies to lipoprotein lipase (anti-LPL) associated with dyslipoproteinemia prompted us to analyze the association of anti LPL with clinical and serologic features in patients with systemic lupus erythematosus (SLE) and its link to markers of inflammation that are known to be involved in atherogenesis. METHODS: Enzyme-linked immunosorbent assay was used to test for the presence of anti-LPL antibodies in 66 consecutive patients with SLE. Clinical and laboratory evaluation, including a fasting lipid profile, autoantibody screening, an assessment for markers of inflammation (C-reactive protein [CRP], erythrocyte sedimentation rate [ESR]), and the SLE Disease Activity Index (SLEDAI) were performed at the time of inclusion in the study. Exclusion criteria were any conditions that affect the lipid profile. SLE patients were categorized into 2 groups according to detection of these anti-LPL antibodies, as follows: anti-LPL+ and anti-LPL-. RESULTS: Anti-LPL antibody IgG was detected in 25 SLE patients (37.8%). Triglyceride levels were significantly higher in the anti-LPL+ group (112.4 +/- 50.2 versus 89.9 +/- 54.5 mg/dl in the anti-LPL- group; P = 0.033), but no significant differences between the 2 groups were detected for total, high-density lipoprotein, and low-density lipoprotein cholesterol levels. A higher frequency of elevated CRP levels and ESRs was observed in the anti-LPL+ group compared with the anti-LPL- group (44% and 17.1%, respectively [P = 0.023] and 52% and 19.5%, respectively [P = 0.013]). Moreover, SLE patients with anti-LPL antibodies also had significantly higher levels of CRP (11.1 +/- 16.4 versus 2.4 +/- 2.6 mug/ml; P = 0.036) and higher ESRs (33.4 +/- 29.8 versus 16.5 +/- 11.8 mm/hour; P = 0.020). Anti-LPL titers had a significant positive correlation with the CRP level (r = 0.56, P < 0.001), the ESR (r = 0.55, P < 0.001), the SLEDAI score (r = 0.45, P < 0.001), anti-double-stranded DNA (anti-dsDNA; r = 0.52, P < 0.001), and anticardiolipin IgG antibodies (r = 0.25, P = 0.04), and a significant negative correlation was detected with total hemolytic complement activity (CH100) (r = -0.34, P = 0.005). Reinforcing these findings, multiple regression analysis also revealed a significant association of anti-LPL with the CRP level (P = 0.025) and anti-dsDNA (P < 0.001). Importantly, a comparison of positive and negative anti-dsDNA sera revealed similar mean CRP levels (P = 0.56) and ESRs (P = 0.102), contrasting with the SLEDAI score (P = 0.004) and CH100 (P = 0.008). CONCLUSION: These data support the link between inflammation, immune response, and dyslipoproteinemia in SLE, introducing anti LPL as a possible new player that may ultimately help in understanding the complex events of atherogenesis in this disease. PMID- 15529372 TI - Increased prevalence of squamous intraepithelial lesions in systemic lupus erythematosus: association with human papillomavirus infection. AB - OBJECTIVE: To compare the prevalence of abnormal Pap smears in patients with systemic lupus erythematosus (SLE) with that in a large group of healthy controls, and to determine whether SLE itself is an independent risk factor. The association of human papillomavirus (HPV) infection and the use of immunosuppressive agents with abnormal Pap smears in SLE was also assessed. METHODS: Eighty-five SLE patients participated in this cross-sectional study. A sample of cervical cells was collected from each patient for routine cytologic examination. HPV was typed by restriction and sequencing analysis. A structured questionnaire was administered to the subjects to ascertain the possible behavioral and biologic risk factors associated with cervical atypia. Data on 2,080 healthy female subjects were retrieved for comparison. RESULTS: The mean (+/-SD) age of SLE patients and controls was 42 +/- 9 years and 44 +/- 10 years, respectively. The prevalence of abnormal Pap smears was significantly increased in SLE patients compared with controls (16.5% versus 5.7%). The prevalence of squamous intraepithelial lesions was increased approximately 6-fold in SLE patients (11.8%) compared with controls (2.0%). SLE itself remained an independent risk factor for abnormal Pap smears (odds ratio 3.5, 95% confidence interval 1.8-6.9). The overall prevalence of HPV infection in SLE patients and controls was 11.8% and 7.3%, respectively. However, 10.6% of SLE patients were infected with at least 1 high-risk type of HPV, compared with 4.2% of controls. Multiple infections were also more common in SLE patients than in controls (4.7% versus 1.1%). There were no significant differences in the use of immunosuppressive agents between SLE patients with normal Pap smears and those with abnormal Pap smears. CONCLUSION: Abnormal Pap smears were more common among SLE patients than controls, even after adjusting for HPV status. SLE-associated immunosuppression increases susceptibility to high-risk HPV infection and multiple HPV infections. The use of immunosuppressant agents was not associated with abnormal Pap smears. PMID- 15529373 TI - Use of 3H-glucosamine and 35S-sulfate with cultured human chondrocytes to determine the effect of glucosamine concentration on formation of chondroitin sulfate. AB - OBJECTIVE: To determine whether addition of glucosamine will stimulate synthesis of chondroitin sulfate by cultures of human chondrocytes, and to compare the relative contribution of endogenous glucosamine to exogenous glucosamine in forming chondroitin sulfate. METHODS: Cultured human chondrocytes were incubated with (35)S-sulfate and various amounts of glucosamine to determine whether any incremental formation of chondroitin (35)S-sulfate occurred. Similarly, chondrocytes incubated with variable concentrations of (3)H-glucosamine were examined to determine how much the incorporation into (3)H-chondroitin sulfate was diluted by provision of endogenous glucosamine that was derived by metabolism from glucose. RESULTS: No stimulation of chondroitin (35)S-sulfate synthesis was found at concentrations of glucosamine up to 1 mM, a concentration of (3)H glucosamine well above the concentrations that could be presented to cartilage after ingestion of advertised amounts of glucosamine. Furthermore, there was significant dilution of exogenous glucosamine by endogenous glucosamine provided by metabolism from glucose. CONCLUSION: The results indicate that exogenous glucosamine does not stimulate chondroitin sulfate synthesis by human chondrocytes. Furthermore, the cells have the capacity to form amounts of glucosamine from glucose far in excess of that provided from exogenous sources, except at concentrations greater than could possibly be achieved with oral administration of glucosamine. PMID- 15529374 TI - Prolonged bleeding time and lupus anticoagulant: a second paradox in the antiphospholipid syndrome. AB - OBJECTIVE: Systemic lupus erythematosus and antiphospholipid syndrome (APS) often appear concomitantly. Lupus nephritis and antiphospholipid antibody-related ischemic nephropathy cannot be distinguished clinically, although their etiology and treatment differ greatly. Examination of a renal biopsy sample is therefore essential in order to provide the correct treatment. We have observed that patients with lupus anticoagulant (LAC), a serologic marker for APS, often have a prolonged bleeding time, which is a contraindication for performing a percutaneous renal biopsy. We undertook this study to evaluate systematically the bleeding time in 27 consecutive patients. METHODS: The bleeding time was measured in 27 patients who were persistently positive for LAC and who were not exposed to aspirin or nonsteroidal antiinflammatory drugs. Platelet function and von Willebrand factor (vWF) parameters were subsequently assessed in patients with a prolonged bleeding time. RESULTS: Twenty-one of 27 patients (78%) had a prolonged bleeding time despite a normal platelet count in the majority of patients (81%). Platelet functioning and vWF parameters in these 21 patients were normal, except for those in 1 patient with a mild storage pool disease. CONCLUSION: With this study, we introduce yet another paradox in the phenomenon of APS. Although a prolonged bleeding time is generally accepted to be a sign of defective primary hemostasis, LAC is associated with thrombosis. Further studies are needed to elucidate the mechanism behind this disturbance of primary hemostasis. PMID- 15529375 TI - Production of endogenous antibiotics in articular cartilage. AB - OBJECTIVE: Defensins are broad-spectrum antimicrobial peptides that are components of innate immunity. To date, only epithelial surfaces and blood cells have been shown to produce these cationic peptides in bactericidal concentrations when challenged with microorganisms or inflammatory cytokines. Infections caused by gram-negative pathogens occur only infrequently in association with joint surgery. The present study was undertaken to investigate whether this may be explained by intraarticular production of gram-negative-specialized antimicrobial peptides. METHODS: Healthy articular cartilage and cultured T/C-28a2 chondrocytes were assessed, by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry, for expression of various antimicrobial peptides. The expression of human beta-defensin 2 (HBD-2) was studied in cultured chondrocytes after exposure to bacterial supernatants and proinflammatory cytokines and was assayed by real-time RT-PCR and immunoblot analysis. A septic arthritis mouse model was used to investigate the regulation of the murine homolog of HBD-2 in articular cartilage after bacterial inoculation. RESULTS: Healthy articular cartilage and T/C-28a2 chondrocytes were able to produce different antimicrobial peptides. After exposure to gram-negative bacteria and proinflammatory cytokines, expression of cartilage-derived HBD-2 strongly increased. Immunoblot analysis revealed up-regulation of the gram-negative-specialized HBD-2 in microbicidal doses. Immunohistochemistry analysis revealed induction of the murine homolog of HBD-2 in vivo after intraarticular injection of bacteria. CONCLUSION: This study demonstrated a previously unrecognized function of human chondrocytes. In addition to its biomechanical properties, articular cartilage has the ability to produce antimicrobial substances when challenged with microorganisms. The expression of HBD-2 in microbicidal doses suggests that antimicrobial peptides may contribute to host defense mechanisms in articular joints. PMID- 15529376 TI - Role of NF-kappaB transcription factors in antiinflammatory and proinflammatory actions of mechanical signals. AB - OBJECTIVE: The mechanisms by which chondrocytes convert biomechanical signals into intracellular biochemical events are not well understood. In this study, we sought to determine the intracellular mechanisms of the magnitude-dependent actions of mechanical signals. METHODS: Chondrocytes isolated from rabbit articular cartilage were grown on flexible membranes. Cells were subjected to cyclic tensile strain (CTS) of various magnitudes in the presence or absence of interleukin-1beta (IL-1beta), which was used as a proinflammatory signal for designated time intervals. The regulation of NF-kappaB was measured by reverse transcriptase-polymerase chain reaction, electrophoretic mobility shift assay, and immunofluorescence. RESULTS: CTS of low magnitudes (4-8% equibiaxial strain) was a potent inhibitor of IL-1beta-dependent NF-kappaB nuclear translocation. Cytoplasmic retention of NF-kappaB and reduction of its synthesis led to sustained suppression of proinflammatory gene induction. In contrast, proinflammatory signals generated by CTS of high magnitudes (15-18% equibiaxial strain) mimicked the actions of IL-1beta and induced rapid nuclear translocation of NF-kappaB subunits p65 and p50. CONCLUSION: Magnitude-dependent signals of mechanical strain utilize the NF-kappaB transcription factors as common elements to abrogate or aggravate proinflammatory responses. Furthermore, the intracellular events induced by mechanical overload are similar to those that are initiated by proinflammatory cytokines in arthritis. PMID- 15529377 TI - Combination of infliximab and methotrexate therapy for early rheumatoid arthritis: a randomized, controlled trial. AB - OBJECTIVE: To compare the benefits of initiating treatment with methotrexate (MTX) and infliximab (anti-tumor necrosis factor alpha [anti-TNFalpha] monoclonal antibody) with those of MTX treatment alone in patients with rheumatoid arthritis (RA) of < or =3 years' duration. METHODS: RA patients were eligible if they had active disease and no prior treatment with MTX or a TNFalpha inhibitor. One thousand forty-nine patients were randomly assigned in a 4:5:5 ratio to 3 treatment groups: MTX-placebo, MTX-3 mg/kg infliximab, and MTX-6 mg/kg infliximab. MTX dosages were rapidly escalated to 20 mg/week, and infliximab or placebo infusions were given at weeks 0, 2, and 6, and every 8 weeks thereafter through week 46. RESULTS: At week 54, the median percentage of American College of Rheumatology improvement (ACR-N) was higher for the MTX-3 mg/kg infliximab and MTX-6 mg/kg infliximab groups than for the MTX-placebo group (38.9% and 46.7% versus 26.4%, respectively; P < 0.001 for both comparisons). Patients in the MTX 3 mg/kg infliximab and MTX-6 mg/kg infliximab groups also showed less radiographic progression than those receiving MTX alone (mean +/- SD changes in van der Heijde modification of the total Sharp score at week 54: 0.4 +/- 5.8 and 0.5 +/- 5.6 versus 3.7 +/- 9.6, respectively; P < 0.001 for each comparison). In addition, physical function improved significantly more in the MTX-3 mg/kg infliximab and MTX-6 mg/kg infliximab groups than in the MTX-placebo group. Infliximab therapy was associated with a significantly higher incidence of serious infections, especially pneumonia. CONCLUSION: For patients with active RA in its early stages, combination therapy with MTX and infliximab provides greater clinical, radiographic, and functional benefits than treatment with MTX alone. PMID- 15529378 TI - Prognostic importance of low body mass index in relation to cardiovascular mortality in rheumatoid arthritis. AB - OBJECTIVE: Various etiologic mechanisms have been implicated in the observed increase in cardiovascular mortality in rheumatoid arthritis (RA). Body mass index (BMI) is associated with cardiovascular mortality in the general population. This study compared the effect of BMI on cardiovascular mortality in a population-based cohort of subjects with RA with that in a cohort of individuals without RA from the same population. METHODS: The RA cohort comprised all members of an incidence cohort of Rochester, Minnesota residents ages > or =18 years who were first diagnosed with RA (by the American College of Rheumatology 1987 criteria) from 1955 through 1994. An age- and sex-matched comparison cohort of subjects without RA was assembled. Both cohorts were followed up longitudinally through their complete (inpatient, outpatient) medical records beginning at age 18 years and continuing until death, migration, or January 1, 2001, and the details of weight and height changes during this period were recorded. High BMI was defined as a BMI >30 kg/m(2) and low BMI as <20 kg/m(2). Cox regression models were used to estimate the effect of BMI on cardiovascular mortality after accounting for traditional cardiac risk factors and malignancies. RESULTS: RA subjects with low BMI at incidence had a significantly higher risk of cardiovascular death (hazard ratio [HR] 3.34, 95% confidence interval [95% CI] 2.23-4.99) compared with non-RA subjects with normal BMI, after adjusting for age, sex, personal cardiac history, smoking status, and presence of diabetes, hypertension, and malignancies. RA subjects with normal BMI at incidence who experienced low BMI during followup also had a higher risk of cardiovascular death (HR 2.09, 95% CI 1.50-2.92) when compared with non-RA subjects who maintained normal BMI throughout followup. CONCLUSION: Among patients with RA, low BMI is associated with a significantly increased risk of cardiovascular death. PMID- 15529379 TI - Effect of etanercept on iritis in patients with ankylosing spondylitis. PMID- 15529380 TI - Criteria for steroid-sparing ability of interventions in systemic lupus erythematosus: report of a consensus meeting. PMID- 15529381 TI - Effect of hypoxia and reoxygenation on gene expression and response to interleukin-1 in cultured articular chondrocytes. AB - OBJECTIVE: To determine the effects of hypoxia and reoxygenation on the metabolism of chondrocytes and their response to interleukin-1beta (IL-1beta). The study included activation of hypoxia-inducible factor 1 (HIF-1), NF-kappaB, and activator protein 1 (AP-1) transcription factors, expression of matrix components and metalloproteases and transforming growth factor beta (TGFbeta) and TGFbeta receptors, and production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)). METHODS: Bovine articular chondrocytes (BACs) were cultured to confluency in either 5% O(2) (hypoxia) or 21% O(2) (normoxia) in media supplemented with 10% fetal calf serum (FCS). BACs were preincubated for 18 hours in media with 1% FCS only and then incubated for 24 hours in the presence of IL 1beta. For reoxygenation experiments, cells were treated in the same way in 5% O(2), except that cultures were transferred to normal atmospheric conditions and used after 4 hours for RNA extraction or after 30 minutes for cytoplasmic or nuclear protein extraction. RESULTS: In hypoxic and reoxygenated chondrocytes, we observed strong DNA binding of HIF-1. IL-1beta-induced DNA binding of NF-kappaB and AP-1 was significantly higher in hypoxic and reoxygenated cultures than in normoxia. Greater activation of the MAPKs was also observed with IL-1beta treatment in hypoxia compared with normoxia. Steady-state levels of type II collagen and aggrecan core protein messenger RNA (mRNA) were decreased by IL 1beta in all instances. Matrix metalloprotease 1 (MMP-1) and MMP-3 mRNA were increased by IL-1beta in normoxia and hypoxia, whereas only MMP-3 mRNA was enhanced in reoxygenated cultures. The MMP-2 mRNA level was not significantly affected by IL-1beta in normoxia or hypoxia, whereas it was enhanced in reoxygenated cultures. MMP-9 mRNA was dramatically decreased by IL-1beta only in low oxygen tension. Tissue inhibitor of metalloproteinases 1 (TIMP-1) message was significantly enhanced by the cytokine in most instances, whereas TIMP-2 message was markedly decreased by IL-1beta in reoxygenated cultures. Stimulation of TGFbeta1 expression by IL-1beta was observed only in normal atmospheric conditions. One of the more striking findings of the study was the greater stimulating effect of IL-1beta on NO production observed in hypoxia, which was much higher than in normoxia, whereas the reverse was observed for IL-1beta induced PGE(2) production. CONCLUSION: Oxygen level and reoxygenation stress significantly modulate gene expression and the response of articular chondrocytes to cytokines such as IL-1beta. In hypoxic conditions, which mimic the in vivo condition of cartilage, the effects of IL-1beta on both synthesis and degradative processes are significantly different from those in normoxia, conditions that are unlikely encountered by chondrocytes in a normal state. In low oxygen tension, high IL-1beta-induced NO production is associated with a significant decrease in PGE(2) synthesis. These data should influence our concept of the role of oxygen in the pathophysiology of joint disease and may help define the best conditions in which to develop bioartificial cartilage. PMID- 15529382 TI - Bone edema and synovial inflammation: comment on the editorial by Kirwan. PMID- 15529383 TI - The American College of Rheumatology response criteria for systemic lupus erythematosus clinical trials: measures of overall disease activity. AB - OBJECTIVE: Improved standards for the evaluation of therapeutic interventions in systemic lupus erythematosus (SLE) are needed. The purpose of this study by a committee of the American College of Rheumatology was to define clinically meaningful improvement, no change, or worsening in 6 existing clinical measures of SLE disease activity. This represents an important step in a disease in which some organ symptoms get better and others get worse. It is intended to help investigators develop sample size estimates based on meaningful effect sizes and to gauge the clinical relevance of any observed change in disease activity. METHODS: Medical records from 310 patients drawn from 3 sources were abstracted into a standard format. Each vignette included clinical and laboratory data obtained during 2-3 visits. Ratings on the following 6 instruments were obtained for the same patients during the visit or retrospectively: the British Isles Lupus Assessment Group (BILAG), the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), the revised Systemic Lupus Activity Measure (SLAM-R), the European Consensus Lupus Activity Measure (ECLAM), the Safety of Estrogens in Lupus Erythematosus: National Assessment (SELENA)-SLEDAI, and the Responder Index for Lupus Erythematosus (RIFLE). From this pool of vignettes, 5 common vignettes and 10 randomly selected vignettes were rated through a secure Web site by 88 international experts on SLE. The experts, who were blinded to the activity measure scores, were asked to rate each patient's clinical condition as worsened, improved, or unchanged relative to the previous visit. These ratings were transformed by statistical procedures into performance characteristic curves that related a change on a particular SLE activity measure to the physicians' agreement on whether that patient had worsened, improved, or remained the same clinically. These were discussed by the committee members, who were blinded to the actual instrument used. The committee then voted on what level of expert agreement would be used to determine clinically meaningful change. RESULTS: The physician ratings on the 5 common vignettes revealed considerable variation in their clinical appraisals. Overall, the 6 SLE activity measures showed excellent separation of clinical conditions as being worsened, improved, or the same. The committee voted to take 70% agreement by physicians as the point on the performance characteristic curves at which meaningful change in a score could be identified. For each instrument, we computed the units of change required to indicate improvement or worsening. CONCLUSION: To our knowledge, these are the first response criteria in any disease where a clinically relevant change has been determined a priori and mapped to standardized measures. This criterion should aid the clinical evaluation of new therapies, improve comparability between trials, and facilitate innovative trial designs. PMID- 15529384 TI - Lectin-like oxidized low-density lipoprotein receptor 1 mediates matrix metalloproteinase 3 synthesis enhanced by oxidized low-density lipoprotein in rheumatoid arthritis cartilage. AB - OBJECTIVE: To investigate for the presence of oxidized low-density lipoprotein (ox-LDL) and lectin-like oxidized LDL receptor 1 (LOX-1) in cartilage specimens from rheumatoid arthritis (RA) joints and to determine whether the interaction of ox-LDL with LOX-1 can induce matrix metalloproteinase 3 (MMP-3) in articular cartilage explant culture. METHODS: Human articular cartilage specimens obtained from patients with RA, osteoarthritis (OA), and femoral neck fractures were examined for LOX-1 and ox-LDL by confocal fluorescence microscopy. The association between ox-LDL and LOX-1 was evaluated by immunofluorescence analysis. Articular cartilage specimens from patients with femoral neck fractures were incubated with ox-LDL, with or without preincubation with neutralizing anti LOX-1 antibody. MMP-3 synthesis by chondrocytes in explant cartilage was evaluated by immunofluorescence, and protein secretion into conditioned medium was monitored by immunoblotting and enzyme-linked immunosorbent assay. RESULTS: The majority of the RA chondrocytes stained positively with both anti-LOX-1 and anti-ox-LDL antibodies; however, no positive cells were found in OA and normal cartilage specimens. Anti-LOX-1 antibody suppressed the binding of DiI-labeled ox LDL to chondrocytes in explant culture, suggesting that the interaction was mediated by LOX-1. In contrast to native LDL, ox-LDL induced MMP-3 synthesis by articular chondrocytes in association with the induction of LOX-1, which resulted in enhanced secretion of MMP-3 into the culture medium. Anti-LOX-1 antibody reversed ox-LDL-stimulated MMP-3 synthesis to control levels. CONCLUSION: Ox-LDL, principally mediated by LOX-1, enhanced MMP-3 production in articular chondrocytes. Increased accumulation of ox-LDL with elevated expression of LOX-1 in RA cartilage indicates a specific role of the receptor-ligand interaction in cartilage pathology in RA. PMID- 15529385 TI - Enhancement of collagen-induced arthritis in mice genetically deficient in extracellular superoxide dismutase. AB - OBJECTIVE: To examine the influence of superoxide on the severity of collagen induced arthritis (CIA) in mice. METHODS: CIA was induced in DBA/1J mice lacking the extracellular superoxide dismutase (EC-SOD) gene (knockout [KO]) and in normal DBA/1J mice (wild-type [WT]). RESULTS: The clinical disease activity score was significantly higher in EC-SOD-KO mice than in WT mice between days 36 and 53, and the histologic scores for joint damage on day 53 increased 2-fold or more in the EC-SOD-KO mice. There were no significant differences between the 2 groups of mice in proliferation indices of spleen or lymph node cells in vitro after stimulation with type II collagen. Although both IgG1 and IgG2a anticollagen antibody levels increased in both groups of mice between days 21 and 53, there were no significant differences between the 2 groups. Lipopolysaccharide stimulated spleen cells from EC-SOD-KO mice produced greater levels of tumor necrosis factor alpha (TNFalpha) over 48 hours in culture compared with cells from WT mice. Increased steady-state levels of messenger RNA (mRNA) for interferon-gamma (IFNgamma), TNFalpha, and interleukin-1beta (IL-1beta), and lower levels of IL-1 receptor antagonist (IL-1Ra) mRNA were present in the joints of the EC-SOD-KO mice compared with the WT mice. CONCLUSION: The absence of EC SOD leads to more severe CIA, which may be accompanied by enhanced production of the proinflammatory cytokines IFNgamma, TNFalpha, and IL-1beta, and decreased production of the antiinflammatory cytokine IL-1Ra in the joints. PMID- 15529386 TI - Exploring complement activation to develop biomarkers for systemic lupus erythematosus. PMID- 15529387 TI - Passive transfer of Sjogren's syndrome IgG produces the pathophysiology of overactive bladder. AB - OBJECTIVE: The presence, in patients with primary and secondary Sjogren's syndrome (SS), of autoantibodies that acutely inhibit M(3) muscarinic receptor (M3R)-mediated bladder contractions is difficult to reconcile with the fact that symptoms of detrusor overactivity and other features of cholinergic hyperresponsiveness occur in this disease. This study was undertaken to examine the in vivo effects of these autoantibodies on bladder function by examining bladder responsiveness and compliance following passive transfer of patient IgG to mice. METHODS: Contractile responses of isolated bladder strips both to the muscarinic agonist carbachol and to electrically evoked acetylcholine release were measured 48 hours after injection of mice with patient or control IgG. A whole bladder assay with intact neuronal pathways was developed to assess bladder wall compliance on filling cystometry. Expression of M3R in bladders from IgG injected mice was assessed by immunohistochemistry. RESULTS: Passive transfer of SS IgG with inhibitory anti-M3R activity produced a paradoxical increase in contractile responses of detrusor strips to cholinergic stimulation. Cystometry of whole bladders revealed a corresponding decrease in bladder wall compliance and phasic detrusor contractions upon filling, replicating the urodynamic features of an overactive bladder. The features of cholinergic hyperresponsiveness were associated with increased postsynaptic M3R expression and were reproduced by injecting mice with a rabbit antibody against the second extracellular loop of M3R. CONCLUSION: These findings are consistent with the notion that there is initial inhibition of parasympathetic neurotransmission by antagonistic autoantibodies to M3R, which produces a compensatory increase in M3R expression in vivo. The enhanced cholinergic responses during bladder distention result in detrusor overactivity. We conclude that the overactive bladder associated with SS is an autoantibody-mediated disorder of the autonomic nervous system, which may be part of a wider spectrum of cholinergic hyperresponsiveness. PMID- 15529388 TI - Histologic analysis of renal leukocyte infiltration in antineutrophil cytoplasmic antibody-associated vasculitis: importance of monocyte and neutrophil infiltration in tissue damage. AB - OBJECTIVE: The histopathologic lesions in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) have been studied extensively, but the exact composition of the cellular infiltrate is unclear. We undertook this study to analyze renal leukocyte infiltration and the cellular distribution within glomeruli and interstitium in 65 renal biopsy samples obtained from patients newly diagnosed as having AAV. METHODS: Renal cellular tissue infiltration was assessed with an immunoperoxidase method. Furthermore, the infiltrating cell types were correlated with clinical and histopathologic data. RESULTS: The predominant interstitial infiltrating cells were T lymphocytes, while monocytes and, to a lesser extent, granulocytes constituted the dominant infiltrating cell types in glomeruli. Interestingly, lymphocyte infiltration was predominantly periglomerular, especially around glomeruli with sclerosis or heavy crescent formation, while interstitial monocyte and neutrophil infiltration was diffusely distributed over the interstitial tissue. A significant correlation was found for the glomerular infiltration of CD68-positive macrophages with the presence of glomerular necrosis as well as with the number of glomeruli with crescents (P < 0.0001 and P = 0.005, respectively). No correlation was found for interstitial fibrosis with the infiltration of any leukocyte subset. Furthermore, a significant correlation was found for the interstitial as well as for the glomerular infiltration of CD68-positive macrophages with serum creatinine concentration at the time of biopsy (P = 0.001 and P = 0.006, respectively). CONCLUSION: These data underscore a major role of monocytes in addition to neutrophils in the tissue damage of AAV. PMID- 15529389 TI - The effect of dehydroepiandrosterone on lumbar spine bone mineral density in patients with quiescent systemic lupus erythematosus. AB - OBJECTIVE: Because dehydroepiandrosterone (DHEA) is an adrenal steroid hormone with weak intrinsic androgenic properties that can be converted in peripheral tissues into more potent sex hormones, one might expect a positive effect of DHEA on bone mineral density (BMD). We evaluated the effects on lumbar BMD of oral DHEA, 200 mg/day, for 1 year in female patients with quiescent systemic lupus erythematosus (SLE). METHODS: The study subjects were 60 women with SLE. All participants gave informed consent to participate in a double-blind, placebo controlled study on the effects of DHEA on fatigue and general well-being. BMD was measured with dual-energy x-ray absorptiometry (DEXA) at baseline and after 12 months. RESULTS: Fifty-eight patients (mean age 42.6 years) could be evaluated; 2 patients (both in the DHEA group) refused to undergo DEXA a second time. In premenopausal women, DHEA did not influence BMD significantly. There was a significant increase in BMD with use of DHEA in postmenopausal women who were not receiving bisphosphonates or estrogen-containing medications. This increase was not observed in the group receiving placebo. CONCLUSION: In premenopausal women with quiescent SLE, use of DHEA does not have a significant effect on BMD. DHEA may increase BMD in postmenopausal SLE patients if they are not already protected from bone loss by use of estrogens or bisphosphonates. Small numbers, due to the absence of stratification for menopausal status, and the use of antiresorptive agents at randomization preclude firmer conclusions based on the results of this study. PMID- 15529390 TI - First reported pediatric case of systemic lupus erythematosus associated with prolactinoma. AB - There is much interest in the possibility that prolactin influences disease activity in systemic lupus erythematosus (SLE). We present the first reported pediatric case of prolactinoma associated with SLE, in a 13-year-old white female. The diagnosis of SLE was based on the presence of arthritis, antinuclear antibodies, and double-stranded DNA, and a chest radiograph showing pleural fluid. The diagnosis of pituitary prolactinoma was based on the histologic features and the presence of amenorrhea, galactorrhea, and an elevated serum prolactin level. Neurosurgical resection and medical therapy with bromocriptine mesylate were independently associated with decreased prolactin levels, loss of arthritis, and reduced levels of inflammatory mediators. PMID- 15529391 TI - Cardiovascular risk factors in women with and without rheumatoid arthritis. AB - OBJECTIVE: The risk of cardiovascular disease (CVD) is increased in patients with rheumatoid arthritis (RA). The objective of this study was to examine the distribution of known CVD risk factors and biomarkers of CVD in women with and without RA. METHODS: This study included two components: an examination of clinical CVD risk factors among women participating in the Nurses' Health Study, a prospective longitudinal cohort, and an analysis of CVD biomarkers among a subgroup of women from this cohort who provided a blood specimen in 1989 (biospecimen cohort). Data regarding clinical risk factors for CVD were collected in 1990 by mailed questionnaire. The diagnosis of RA was confirmed through a structured medical record abstraction. We compared clinical risk factors for CVD and biomarkers of CVD between women with and without RA, adjusting for age, body mass index (BMI), smoking status, and menopause status. RESULTS: Women with RA (n = 287) were significantly more likely than women without RA (n = 87,019) to report no alcohol use (48.2% versus 39.4%) and past cigarette smoking (47.8% versus 38.0%). No significant differences between these groups were observed for current smoker status, BMI, regular aspirin use, diabetes, hypertension, physical activity, and family history of early myocardial infarction. In the biospecimen cohort (69 RA cases and 491 controls), the levels of several inflammatory biomarkers linked to CVD were significantly elevated in women with RA, including CRP, fibrinogen, sICAM-1, sTNFRI, sTNFRII, and osteoprotegerin. Levels of total cholesterol, low-density lipoprotein, triglycerides, apolipoprotein B, and Lp(a) were similar between groups. Levels of homocysteine were similar, but vitamin B(12) was significantly higher among women with RA than among the controls. CONCLUSION: In women participating in the Nurses' Health Study, most traditional CVD risk factors were similar between those who had RA and those who did not. However, as expected, biomarkers of inflammation associated with CVD were generally elevated in women with RA. PMID- 15529392 TI - The presence of citrullinated proteins is not specific for rheumatoid synovial tissue. AB - OBJECTIVE: Antibodies directed toward citrullinated proteins (e.g., anti-cyclic citrullinated peptide antibodies) are highly specific for rheumatoid arthritis (RA) and are produced locally at the site of inflammation. Although the presence of citrullinated proteins in rheumatoid synovium has been described in the literature, it is uncertain whether their presence is specific for RA. The present study was undertaken to investigate this. METHODS: The local production of the anti-citrullinated protein antibodies was investigated by comparing the concentration of the antibodies (corrected for the total amount of IgG present) in paired samples of serum and synovial fluid from RA patients. The presence of citrullinated proteins in the synovial tissue was investigated by immunohistochemical analysis of synovial tissue from RA patients and from patients with other arthropathies, using a variety of specific antibodies to citrullinated proteins. RESULTS: In RA patients, anti-citrullinated protein antibodies constituted a 1.4-fold higher proportion of IgG in synovial fluid compared with serum, which is indicative of a local production of the antibodies. Immunohistochemical staining of citrullinated proteins was observed in the lining layer, the sublining layer, and in extravascular fibrin deposits in inflamed synovial tissue from RA as well as non-RA patients. CONCLUSION: The presence of citrullinated proteins in the inflamed synovium is not specific for RA, but rather, it may be an inflammation-associated phenomenon. The high specificity of the anti-citrullinated protein antibodies is, therefore, most likely the result of an abnormal humoral response to these proteins. PMID- 15529395 TI - Studies on the homogeneous acetylation of cellulose in the novel solvent dimethyl sulfoxide/tetrabutylammonium fluoride trihydrate. AB - The novel solvent dimethyl sulfoxide (DMSO)/tetrabutylammonium fluoride trihydrate (TBAF . 3H(2)O) was studied for acetylation of linters cellulose. In order to control the degree of substitution (DS), acetylation of the macromolecule was carried out at different reaction time and temperature, molar ratio of reactants, as well as under variation of the concentration of TBAF . 3H(2)O in solution. Cellulose acetate (CA) was accessible with DS ranging from 0.43 to 2.77. The change in concentration of TBAF . 3H(2)O in DMSO showed a strong influence on DS. The most appropriate reaction conditions for acetylation of linters cellulose regarding maximal DS were evaluated. The structure of the CA was characterized by means of FTIR and NMR spectroscopy. The solubility of the CA depends not only on the DS but also on the reaction conditions applied, indicating a different distribution of acetate moieties both within and between polymer chains. PMID- 15529396 TI - Production of D-lactic acid by bacterial fermentation of rice starch. AB - D-Lactic acid was synthesized by the fermentation of rice starch using microorganisms. Two species: Lactobacillus delbrueckii and Sporolactobacillus inulinus were found to be active in producing D-lactic acid of high optical purity after an intensive screening test for D-lactic acid bacteria using glucose as substrate. Rice powder used as the starch source was hydrolyzed with a combination of enzymes: alpha-amylase, beta-amylase, and pullulanase to obtain rice saccharificate consisting of maltose as the main component. Its average gross yield was 82.5%. Of the discovered D-lactic acid bacteria, only Lactobacillus delbrueckii could ferment both maltose and the rice saccharificate. After optimizing the fermentation of the rice saccharificate using this bacterium, pilot scale fermentation was conducted to convert the rice saccharificate into D-lactic acid with a D-content higher than 97.5% in a yield of 70%. With this yield, the total yield of D-lactic acid from brown rice was estimated to be 47%, which is almost equal to the L-lactic acid yield from corn. The efficient synthesis of D-lactic acid can open a way to the large scale application of high-melting poly(lactic acid) that is a stereocomplex of poly(L lactide) and poly(D-lactide). Schematic representation of the production of D lactic acid starting from brown rice as described here. PMID- 15529397 TI - Tamoxifen-loaded polymeric micelles: preparation, physico-chemical characterization and in vitro evaluation studies. AB - Several samples of polymeric micelles, formed by amphiphilic derivatives of PHEA, obtained by grafting into polymeric backbone of PEGs and/or hexadecylamine groups (PHEA-PEG-C(16) and PHEA-C(16)) and containing different amount of Tamoxifen, were prepared. All Tamoxifen-loaded polymeric micelles showed to increase drug water solubility. TEM studies provided evidence of the formation of supramolecular core/shell architectures containing drug, in the nanoscopic range and with spherical shape. Samples with different amount of encapsulated Tamoxifen were subjected to in vitro cytotoxic studies in order to evaluate the effect of Tamoxifen micellization on cell growth inhibition. All samples of Tamoxifen loaded polymeric micelles showed a significantly higher antiproliferative activity in comparison with free drug, probably attributable to fluidification of cellular membranes, caused by amphiphilic copolymers, that allows a higher penetration of the drug into tumoral cells. To gain preliminary information about the potential use of prepared micelles as Tamoxifen drug delivery systems, studies evaluating drug release ability of micelle systems in media mimicking biological fluids (buffer solutions at pH 7.4 and 5.5) and in human plasma were carried out. These studies, performed evaluating the amount of Tamoxifen that remains in solution as a function of time, showed that at pH 7.4, as well as in plasma, PHEA-C(16) polymeric micelles were able to release lower drug amounts than PHEA-PEG(5000)-C(16) ones, while at pH 5.5, the behavior difference between two kind of micelles was less pronounced. PMID- 15529398 TI - Helical poly(3-methyl-4-vinylpyridine)/amino acid complexes: preparation, characterization, and biocompatibility. AB - Helical poly(3-methyl-4-vinylpyridine) (P3M4VP)/amino acid complexes have been prepared via acid-base reaction of the achiral polymer with D and L amino acids: alanine, leucine, valine, serine and phenylalanine. The circular dichroism (CD) spectra of P3M4VP/D- and L-alanine complexes in CH(3)OH/H(2)O show opposing (near mirror image) Cotton effect signals at 278.4, 274.8 and 270.8 nm, indicating the formation of enantiomeric secondary structures. The formation of the enantiomeric structures is supported by observed [alpha](D)(25) values of -3.0 and +3.0 for the P3M4VP/D-alanine and P3M4VP/L-alanine complexes, respectively. The preparation of helical P3M4VP/amino acid complexes has been carried out in CH(3)OH and H(2)O at pH 1.8 and 2.7. The intensities of the Cotton effect signals were good. For example, for the P3M4VP/L-alanine complexes in CH(3)OH/H(2)O and H(2)O (pH 1.8), the second Cotton effect signal around 275-277 nm show [theta;] values of 49 980 and 79 210 deg . cm(2) . dmol(-1), respectively. The formation of the helical secondary structure is rapid. The acid-base reaction between P3M4VP and L-alanine in CH(3)OH/H(2)O, in 10 min, show a CD spectrum with Cotton effect signals at 274 and 272 nm with [theta] values of 27,000 deg . cm(2) . dmol(-1) and -36,000 deg . cm(2) . dmol(-1), respectively. P3M4VP permits ready conformational reorientation on complexation with amino acids, but once the helical P3M4VP/amino acid complexes are formed, it is stable at room temperature. P3M4VP is not compatible with HeLa ovarian cancer cells, but the helical P3M4VP/amino acid complexes are compatible with HeLa cells. The complexes minimally interfere with the adhesion and growth of HeLa cells on complex surfaces. Helical poly(3-methyl-4-vinylpyridine)/D- and L-alanine complexes support the attachment and growth of HeLa cells. The micrographs shows HeLa cells after three days: left panel: on P3M4VP/L-alanine complex; right panel: on P3M4VP/D-alanine complex. PMID- 15529399 TI - Erythropoietin production rate in phlebotomy-induced acute anemia. AB - OBJECTIVE: To estimate the rate of erythropoietin (EPO) production under physiological, conditions and to examine the regulatory mechanism of EPO production in response to acute phlebotomy-induced anemia. METHODS: Six sheep each underwent two phlebotomies in which the hemoglobin (Hb) was reduced to 3-4 g/dl over 4-5 h. The EPO plasma level, reticulocytes, Hb and EPO clearance were followed by frequent blood sampling. The EPO production rate was determined by a semi-parametric method based on a disposition decomposition analysis that accounts for the nonlinear disposition kinetics of EPO and corrects for time dependent changes in the clearance. RESULTS: The controlled drop in hemoglobin resulted in an abrupt increase in the plasma EPO concentration (peak level 812+/ 40 mU/ml, mean+/-CV%) that was followed by a rapid drop 2-4 days after the phlebotomy at a time when the sheep were still anemic (Hb=4.3+/-16 g/dl). The EPO production rate at baseline was 43+/-52 U/day/kg and the amounts of EPO produced over an 8 day period resulting from the first and second phlebotomy were 2927+/ 40 U/kg and 3012+/-31 U/kg, respectively. CONCLUSIONS: The rapid reduction in the EPO plasma level observed 2-4 days following the phlebotomy cannot be explained solely by the increase in EPO clearance but also by a reduction in EPO production. PMID- 15529400 TI - Cytologic factors associated with prognosis in patients with peripheral adenocarcinoma of the lung measuring 3 cm or less in greatest dimension. AB - BACKGROUND: Recently, peripheral lung adenocarcinomas (PLA) measuring < or = 3 cm in greatest dimension often have been diagnosed using diagnostic radiology. The objective of the current study was to determine which cytologic factors are associated with a favorable outcome and an unfavorable outcome in patients with PLA. METHODS: Imprint smears from 134 patients with PLA were examined. Sixteen cytologic factors, including necrosis, cellular distribution, overlapping of cell clusters, cluster aggregation, cluster size, cluster thickness, nuclear irregularity, nuclear size, variation in nuclear size, multinucleated cells, intranuclear inclusions, type of intranuclear inclusions, appearance of nucleoli, eosinophilic nucleoli, multinucleoli, and mitosis, were evaluated using univariate and multivariate analyses. A counting method was used to determine the prognosis for individual patients. RESULTS: In the univariate analysis, a cluster size that measured > or = 831 microm in short dimension (P = 0.0011), moderate or severe nuclear irregularity (P = 0.0030), > or = 5 multinucleated cells per 100 tumor cells (P = 0.0047), moderate or severe variation in nuclear size (P = 0.0061), medium or large nuclear size (P = 0.0169), and > or = 1 mitotic cell per 100 tumor cells (P = 0.0412) were associated significantly with a poor outcome. In the multivariate analysis, cluster size in short dimension (P = 0.0018), multinucleated cells (P = 0.0066), and nuclear irregularity (P = 0.0310) were found to be independent prognostic factors. CONCLUSIONS: The combination of cytologic features using intraoperative imprint smears, namely, cluster sizes < or = 830 microm in short dimension, < or = 4 multinucleated cells per 100 tumor cells, and mild nuclear irregularity, may provide favorable predictive information in patients with PLA. PMID- 15529403 TI - Direct injection of 96-well organic extracts onto a hydrophilic interaction chromatography/tandem mass spectrometry system using a silica stationary phase and an aqueous/organic mobile phase. PMID- 15529404 TI - Analysis of silymarin extracted from a commercial dosage by combining liquid liquid extraction with negative electrospray tandem mass spectrometry. PMID- 15529405 TI - Proteome analysis of red deer antlers. AB - Deer antlers are the only mammalian organs capable of repeated regeneration. Although antlers are known to develop from pedicles, which arise from antlerogenic cells of cranial periosteum, their developmental process is not fully elucidated. For example, while endocrine and environmental factors influence the antler development, it is still unclear which signaling pathways are involved in the transduction of such stimuli. To study the developmental process of antlers and identify proteins functioning in their growth, we have established proteome maps of red deer (Cervus elaphus) antlers. With two dimensional gel electrophoresis and matrix-assisted laser desorption/ionization mass spectrometry, we analyzed more than 800 protein spots and identified approximately 130 individual proteins derived from the growing tip of antlers. The overall profile of the antler proteome was dissimilar to those of other types of tissue. Also comparison of proteomes derived from proximal bony tissue and the growing tip of antlers revealed substantial differences. Moreover several cell growth or signaling-related proteins are expressed exclusively in the growing tip, suggesting that these proteins function in the growth and differentiation of antlers. Currently, using the antler proteome maps, we are actively searching for the regulatory factor(s) that may control the antler development. PMID- 15529406 TI - Psychrophilicity of Bacillus psychrosaccharolyticus: a proteomic study. AB - Psychrophilicity of Gram-positive bacterium, Bacillus psychrosaccharolyticus was investigated in a proteomic approach. One hundred and thirty-one protein spots were analyzed by electrospray ionization-quadrupole-time of flight-tandem mass spectrometry and identified using an unpublished translated contig database as well as a nonredundant Gram-positive bacteria protein database from NCBI because of the lack of a genome sequence of this organism. Results focused on proteomic behavior of cold-response show that global up-regulation of metabolic functions and protective mechanism by stress responses might play a major role in psychrophilicity of B. psychrosaccharolyticus. PMID- 15529407 TI - Establishment of a near-standard two-dimensional human urine proteomic map. AB - A proteomic map for human urine on two-dimensional (2-D) gels has been developed. Initial studies demonstrated that the urine proteins prepared by conventional methods showed interference and poor reproducibility in 2-D electrophoresis (2 DE). To address this issue, urine samples were dialyzed to remove any interfering molecules. The dialysis of urine proteins and the concentration by lyophilization without fractionation significantly improved the reproducibility and resolution and likely represents the total urine proteins on a 2-D gel. In addition, removing albumin from urine using Affi-Gel Blue helped to identify the low abundant proteins. Using the developed method, we prepared proteins from urine collected from healthy females and males. The large inter- and intra-subject variation in protein profiles on 2-D gels made it difficult to establish a normal human urine proteomic 2-D map. To resolve this problem, urinary proteins were prepared from the pooled urine collected from 20 healthy females and males, respectively. The established male and female urine proteomes separated on 2-D gels were almost identical except for some potential sex-dependent protein spots. We have annotated 113 different proteins on the 2-D gel by peptide mass fingerprinting (PMF). We propose that the established total urine proteome can be used for 2-DE analysis, liquid chromatography-tandem mass spectrometry (LC MS/MS), and identification of novel disease-specific biomarkers. PMID- 15529408 TI - Effects of benzo(a)pyrene on protein expression in Jurkat T-cells. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants of air, water and soil, and are produced by the incomplete combustion of organic materials. The International Agency for Research on Cancer has characterized PAHs as carcinogens. In this study, we investigated the effects of benzo(a)pyrene (B(a)P), which is the most carcinogenic member of the PAHs, on Jurkat cell protein by proteomic analysis. Jurkat cells were treated with various concentrations of B(a)P (0, 2.5, 5, 10, 20 or 40 microM) for 24 or 48 h and 3 (4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H tetrazolium and lactate dehydrogenase assays were carried out to determine cytotoxicity and a Comet assay was used to determinate genotoxicity. The cytotoxicity assays showed that 2.5 microM of B(a)P was the maximal concentration that did not cause any toxicity, but nevertheless, at this level B(a)P produced significant DNA damage in Jurkat cells at 48 h. Proteomic analysis using three different pI ranges and large two-dimensional gel electrophoresis showed 3427 protein spots. A total of 46 (13 up- and 33 down-regulated) proteins were identified as biomarkers of B(a)P and showed dose-dependent expressions in Jurkat T-cell line exposed to B(a)P. Of these, 27 protein spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Two functionally differentiated protein groups were found. The protein group involving apoptosis and tumor suppression were found to be up-regulated, and B(a)P down-regulated enzyme was involved in energy metabolism, DNA synthesis and in cell structure and motility. PMID- 15529409 TI - Two-dimensional electrophoretic analysis reveals that lipid rafts are intact at physiological temperature. AB - Different proteins are found in lipid rafts depending on the isolation method. For example, insulin receptor was predominantly found in lipid raft fractions prepared from HepG2 cells with Brij 35, but were not present in lipid rafts isolated with Triton X-100. In order to assess the effect of detergent type and temperature on raft isolation, raft proteins from HepG2 cells were analyzed by two-dimensional (2-D) electrophoresis. More raft protein spots appeared when rafts were isolated by Brij 35 than by Triton X-100. In addition, more raft proteins were found when isolated at 37 degrees C than at 4 degrees C, indicating that lipid rafts are much more stable at physiological temperature (37 degrees C) in the presence of detergents. Indeed, lipid-modified proteins, such as Src and Lyn, were found in raft fractions even when detergent-resistant rafts were isolated at room or physiological temperature. The 2-D gel profile of raft proteins isolated with detergent-free (high-pH/carbonate) method was considerably similar to that of detergent-resistant raft proteins but contained a greater number of distinct protein spots. Whereas many detergent-resistant raft proteins disappeared upon cellular exposure to methyl-beta-cyclodextrin, high pH/carbonate resistant raft proteins did not, suggesting that many of proteins isolated by high pH/carbonate could be contaminants. Considering these data, we conclude that liquid-ordered state of detergent-resistant lipid rafts is not destroyed at physiological temperature. PMID- 15529410 TI - Identification of the degradome of Isp-1, a major intracellular serine protease of Bacillus subtilis, by two-dimensional gel electrophoresis and matrix- assisted laser desorption/ionization-time of flight analysis. AB - Intracellular serine protease-1 (Isp-1) is a major intracellular serine protease of Bacillus subtilis, whose functions still remain largely unknown. Furthermore, physiological substrates are yet to be determined. To identify Isp-1 substrates, we digested extract obtained from an Isp-1 deficient Bacillus mutant with purified Isp-1 and examined eliminated or decreased spots by two-dimensional gel and matrix-assisted laser desorption/ionization-time of flight analyses. Proteins degraded by Isp-1, termed the Isp-1 degradome, are involved in a variety of cellular functions such as DNA packing, genetic competence, and protein secretion. From the degradome we selected ClpC and EF-Tu as putative Isp-1 substrates and studied their in vitro degradation. ClpC and EF-Tu contain putative cleavage sites for Isp-1. N-terminal sequencing of in vitro proteolytic fragments of ClpC and EF-Tu revealed that these sites are indeed recognized and cleaved by Isp-1. Moreover, the cellular levels of ClpC and EF-Tu were dramatically reduced at the late stationary phase, where the expression level of Isp-1 was greatly increased. These results suggest that the regulated proteolysis of ClpC by Isp-1 plays an important role in the stationary phase adaptive response. This degradomic approach could provide a powerful tool for finding physiological substrates of many proteolytic enzymes whose functions remain to be determined. PMID- 15529413 TI - Identification of target proteins of N-acetylglucosaminyl-transferase V and fucosyltransferase 8 in human gastric tissues by glycomic approach. AB - To understand better the mechanism underlying gastric cancer and search for potential markers for gastric cancer prognosis, the proteomic analysis of gastric cancer tissues was conducted using two-dimensional gel electrophoresis and lectin blot, followed by electrospray ionization-mass spectrometry. These approaches permitted identification of glyco- or putative glycosylated proteins which may be associated with tumorigenesis. The proteins identified include molecules involved in sugar metabolism, signal transduction, proteolysis, and stress, as well as several unknown proteins, which were aberrantly glycosylated as evidenced by the L-phytohemagglutinin blot. PMID- 15529414 TI - Capillary electrophoresis off-line matrix-assisted laser desorption/ionisation mass spectrometry of intact and digested proteins using cationic-coated capillaries. AB - Capillary electrophoresis (CE) was coupled off-line with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS) for the analysis of proteins and peptides. CE fractions were collected directly on a matrix-coated MALDI target, using a sheath-flow interface. Protein adsorption during CE separations was prevented by coating the capillaries with the physically adsorbed, cationic polymer PolyE-323. The CE/MALDI-MS system was used for the analysis of model proteins and peptides at physiological pH as well as analysis of proteins in tear fluid. Moreover, tryptic on-target digestion of the collected protein fractions, with subsequent MALDI-MS and MS/MS peptide analysis, was demonstrated. PMID- 15529415 TI - Atmospheric pressure chemical ionisation liquid chromatography/mass spectrometry of the ultraviolet screening pigment scytonemin: characteristic fragmentations. AB - Atmospheric pressure chemical ionisation reversed-phase high-performance liquid chromatography/multistage mass spectrometry has been used to study the mass spectral fragmentation of the cyanobacterial sheath pigment scytonemin and its reduced counterpart. The two pigments exhibit characteristic fragment ions in their MS2 and MS3 spectra that are of value in confirming the identification of the structures in extracts from natural environments. PMID- 15529417 TI - UV laser ablation of GdCa4O(BO3)3 (GdCOB) investigated by Fourier transform ion cyclotron resonance mass spectrometry. AB - The ions generated by laser ablation (LA) of calcium and gadolinium oxoborate GdCa4O(BO3)3 (GdCOB) were investigated by Fourier transform ion cyclotron resonance mass spectrometry (FTICRMS), a powerful tool for the characterization of ionic species produced by laser interaction with solid material. In order to better understand the matter transfer and the mechanism of thin film growth by pulsed-laser deposition (PLD), cationic and anionic clusters generated by UV laser ablation of GdCOB bulk material were studied. Laser ablation of GdCOB leads to the formation of various cluster ions which result from association of CaO, BO and B2O3 building blocks (BB) with different charge carriers (CC): H+, BO+, GdO+ in positive ion mode, and BO2-, OK-, OH-, Cl-, WO3- in negative ion mode. LA FTICRMS investigations allow us to assign a valence state to each metallic atom included in each BB. A +II chemical state may be associated with calcium and +II and +III ones to boron. UV laser ablation of GdCOB therefore induces reduction processes of boron species in the gas phase. The oxygen reactive atmosphere used during PLD experiments allows the growth of stoichiometric thin films by fixation of oxygen on the ablated species. PMID- 15529416 TI - Matrix-assisted and polymer-assisted laser desorption/ionization time-of-flight mass spectrometric analysis of low molecular weight polystyrenes and polyethylene glycols. AB - Recently, matrices based on oligomers of dioxin and thiophene (polymer-assisted laser desorption/ionization (PALDI)) have been described for mass spectrometric (MS) analysis of low molecular weight compounds (Woldegiorgis A, von Kieseritzky F, Dahlstedt E, Hellberg J, Brinck T, Roeraade J. Rapid Commun. Mass Spectrom. 2004; 18: 841-852). In this paper, we report the use of PALDI matrices for low molecular weight polymers. An evaluation with polystyrene and polyethylene glycol showed that no charge transfer ionization occurs. Ionization is mediated through metal ion adduction. Comparison of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) data for two very low molecular weight polymers with data obtained from size-exclusion chromatography (SEC) revealed a systematic difference regarding mean molecular weight and dispersity. Further, the mass spectra obtained with PALDI matrices had a higher signal-to-noise ratio than the spectra obtained with conventional matrices. For polymers with higher molecular weights (>1500 Da), the conventional matrices gave better performance. For evaluation of the MALDI spectra, three non-linear mathematical models were evaluated to model the cumulative distributions of the different oligomers and their maximal values of Mw, Mn and PDI. Models based on sigmoidal or Boltzmann equations proved to be most suitable. Objective modeling tools are necessary to compare different sample and instrumental conditions during method optimization of MALDI analysis of polymers, since the bias between MALDI and SEC data can be misleading. PMID- 15529418 TI - Rapid determination of five probe drugs and their metabolites in human plasma and urine by liquid chromatography/tandem mass spectrometry: application to cytochrome P450 phenotyping studies. AB - A liquid chromatography/mass spectrometry method, for rapid determination of five cytochrome P450 (CYP) probe drugs and their relevant metabolites in human plasma and urine, is described. The five specific probe substrates/metabolites, caffeine/paraxanthine (CYP1A2), tolbutamide/4 hydroxytolbutamide/carboxytolbutamide (CYP2C9), omeprazole/5-hydroxyomeprazole (CYP2C19), debrisoquine/5-hydroxydebrisoquine (CYP2D6) and midazolam/1' hydroxymidazolam (CYP3A), together with the internal standards (phenacetin and paracetamol), in plasma and urine, were extracted using solid-phase extraction. The chromatography was performed using a C18 column with an isocratic mobile phase consisting of acetonitrile and 0.1% formic acid in water (70:30). The triple-quadrupole mass spectrometer was operated in both positive and negative modes, and multiple reaction monitoring was used for quantification. The method was validated over the concentration ranges 0.05-5 microg/mL for caffeine and paraxanthine, 0.02-2 microg/mL for tolbutamide, 0.1-20 microg/mL for 4 hydroxytolbutamide, carboxytolbutamide, debrisoquine and 5-hydroxydebrisoquine, 5 2500 ng/mL for omeprazole and 5-hydroxyomeprazole, and 1-100 ng/mL for midazolam and 1'-hydroxymidazolam. The intra- and inter-day precision were 0.3-13.7% and 1.9-14.3%, respectively, and the accuracy ranged from 93.5-107.2%. The lower limit of quantification varied between 1 and 100 ng/mL. The present method provides a robust, fast and sensitive analytical tool for the five-probe drug cocktail, and has been successfully applied to a clinical phenotyping study in 16 subjects. PMID- 15529419 TI - A thin chip microsprayer system coupled to Fourier transform ion cyclotron resonance mass spectrometry for glycopeptide screening. AB - A thin polymer microchip was coupled with a Fourier transform ion cyclotron resonance (FTICR) 9.4 T mass spectrometer and the method was optimized in negative ion mode for glycopeptide screening. The interface between the polymer microchip and FTICR mass spectrometer consists of an in-laboratory conceived and designed mounting system that exhibits robust and controllable alignment of the chip toward the inlet of the mass spectrometer. The particular attribute of the polymer chip coupled to the FTICR mass spectrometer, to achieve an increase in ionization efficiency and sensitivity under the premise of high mass accuracy of detection, is highlighted by the large number of major and minor glycopeptide structures detected and identified in highly heterogeneous mixtures obtained from urine matrices. Glycoforms expressing various saccharide chain lengths ranging from tri- to dodecasaccharide, bearing up to three sialic acid moieties, could be detected and assigned based on the accuracy of the mass measurement (average mass deviation below 6 ppm) of their molecular ions. -Thin chipESI-FTICRMS is a potent novel system for glycomic screening of complex mixtures, as demonstrated for identification of singly sialylated O-glycosylated amino acids and peptides from urine matrices, and could be considered for general applicability in the glycoanalytical field. PMID- 15529420 TI - Epidemiology of hypertension. The Budapest Conference. Budapest, Hungary. June 29, 2002. PMID- 15529425 TI - Laboratory diagnosis of large granular lymphocytic leukemia. PMID- 15529426 TI - Remote diagnostics. Cyber service. PMID- 15529427 TI - Hematology external quality assessment in Europe. PMID- 15529428 TI - External hematology quality assurance initiative in Asia. Asian Quality Assurance Survey (AQuAS)Program. PMID- 15529429 TI - Decision rules for hematology review. An international perspective. PMID- 15529430 TI - Decision rules for hematology review. An international perspective. PMID- 15529431 TI - Virtual microscopy. A new tool for hematology education and training. PMID- 15529432 TI - What can be done to prevent errors in the hematology lab? PMID- 15529433 TI - Anemia of chronic disease--pathophysiology and laboratory diagnosis. PMID- 15529434 TI - Diagnostic versus predictive flow cytometry. PMID- 15529435 TI - Minimal residual disease detection by flow cytometry. PMID- 15529436 TI - The left shift: are there granulocytic surface markers that are better indicators? PMID- 15529437 TI - Neoplasias of dentritic cells: are they the counterpart of one or more cell lineages? PMID- 15529438 TI - Laboratory diagnosis of thrombophilia: Why, who, what, when, how to test. PMID- 15529439 TI - Ximelagatran, a new oral anticoagulant--how will it affect laboratory practice? A clinical perspective. PMID- 15529440 TI - Ximelagatran, a new oral anticoagulant--how will it affect laboratory practice? A laboratory perspective. PMID- 15529441 TI - Quality assurance in hemostasis. A European versus North American perspective. PMID- 15529442 TI - College of American pathologists: proficiency testing in coagulation. PMID- 15529443 TI - Gene expression profiling in acute lymphoblastic leukemia (ALL). PMID- 15529444 TI - CML resistance to tyrosine kinase inhibitors: how is the laboratory to tell? PMID- 15529445 TI - Clonality in essential thrombocythaemia. PMID- 15529446 TI - Giant platelet syndromes and the MYH9 mutations. PMID- 15529447 TI - [Chronic inflammatory bowel diseases]. PMID- 15529448 TI - Nevirapine plus zidovudine to prevent mother-to-child transmission of HIV. PMID- 15529449 TI - Recurrent venous thromboembolism in men and women. PMID- 15529450 TI - Recurrent venous thromboembolism in men and women. PMID- 15529451 TI - Recurrent venous thromboembolism in men and women. PMID- 15529452 TI - Recurrent venous thromboembolism in men and women. PMID- 15529453 TI - Recurrent venous thromboembolism in men and women. PMID- 15529454 TI - The antibiotic pipeline. PMID- 15529455 TI - XV International AIDS Conference in Bangkok: antiretroviral therapy. PMID- 15529456 TI - XV International AIDS Conference: women and HIV in the spotlight. PMID- 15529457 TI - Management of community-associated methicillin-resistant Staphylococcus aureus infections in the outpatient setting. PMID- 15529458 TI - Applying motivational enhancement therapy to HIV prevention and care. PMID- 15529459 TI - Motivational interviewing and psychotherapy. PMID- 15529460 TI - Nutrition and HIV/AIDS. PMID- 15529461 TI - The acute effect of ACEI and ARB's upon endothelin in heart failure patients. AB - PURPOSE OF THE STUDY: to compare the acute effect of ACEI and ARB's upon the plasmatic endothelin level in heart failure patients. METHODS: There were studied 30 patients with congestive heart failure, III-IV functional NYHA class, that were not under ACEI or ARB's treatment. In all the patients the endothelinl-21 plasmatic level was determined. After this, 20 patients, representing group I, received a single dose of 1 mg trandolapril, and 10 patients, representing group II, received a single dose of 40 mg telmisartan. After 24 hours, the plasmatic endothelin 1-21 level was determined again. RESULTS: The mean endothelial plasmatic level was similar in both groups (group I: 0.358+/-0.04 fmol/ml; group II: 0.345+/-0.038). After 24 hours, the endothelin level decreased to 0.295+/ 0.03 fmol/ml for group I (p<0.05) and to 0.287+/-0.029 fmol/ml for group II (p<0.05), suggesting that both ACEI and ARB's are equally efficient in decreasing endothelin. The initial endothelinl-21 level was inversely correlated with LVEF (r = -0.989), and the degree of the decrease of endothelinl-21 after both ACEI and ARB's is directly correlated with the initial endothelinl-21 level (r =0.64). CONCLUSION: The acute administration of ACEI and ARB's in heart failure patients decreases the endothelin level, both categories of drugs having the same effects. PMID- 15529462 TI - Diagnosis and management of depression and psychosis in children and adolescents with epilepsy. AB - The neurologic dysfunction underlying epilepsy can predispose patients to psychiatric disorders, and the incidence of both depression and psychosis is increased in people with epilepsy. Depressive disorders are the most frequently recognized psychiatric comorbidities in people with epilepsy, but depression in children can be particularly difficult to recognize. Clinicians need to inquire about not only classic symptoms of depression such as anhedonia but also less obvious symptoms such as unprovoked irritability, unsubstantiated complaints of lack of love from family members, somatic complaints, and problems with concentration and poor school performance. The diagnosis of depressive disorders in children with epilepsy and mental retardation is even more difficult. Physicians need to be alert for the presence of iatrogenic depression, which may result from antiepileptic drugs or epilepsy surgery. People with epilepsy are also at increased risk for psychosis, which can be interictal, postictal, or (rarely) an expression of ictal activity. This psychosis can be related to seizure remission (ie, alternative psychosis) or iatrogenic (eg, related to antiepileptic drugs or following temporal lobectomy). Although both antidepressants and antipsychotic drugs have the potential to lower the seizure threshold and increase seizures, careful drug selection, dosing, and slow titration can minimize this risk, allowing treatment to proceed. PMID- 15529464 TI - Current awareness on yeast. PMID- 15529463 TI - Reactive oxygen and nitrogen species: diagnostic, preventive, and therapeutic values. Proceedings of an international workshop. July 8-12, 2002. St. Petersburg, Russia. PMID- 15529465 TI - Medicare education in housing facilities. AB - Housing professionals who work with older or disabled residents often are asked questions about Medicare-related issues. Many lack the training and education to adequately respond to the questions because their primary responsibilities are not directly related to Medicare. This brief describes a pilot project that tested different methods to educate housing and social services staff about Medicare to better equip them to assist residents with Medicare-related informational needs, and perhaps to better serve as a resource to the broader community. PMID- 15529466 TI - Proceedings of the 7th Annual Meeting of the Society of Biotherapeutic Approaches and the 2nd Annual Meeting of the Japan Research Association for Immunotherapeutics. December 7, 2003. Tokyo, Japan. PMID- 15529467 TI - Health savings accounts: issues and implementation decisions for states. PMID- 15529468 TI - The role of reinsurance in state efforts to expand coverage. PMID- 15529469 TI - Mental health care disparities among youths vary by state. PMID- 15529470 TI - The Constitution at the threshold of life and death: a suggested approach to accommodate an interest in life and a right to die. PMID- 15529471 TI - The parent trap: uncovering the myth of "coerced parenthood" in frozen embryo disputes. PMID- 15529472 TI - Endorsing religion: drug courts and the 12-step recovery support program. PMID- 15529473 TI - A rural perspective regarding regulations implementing Titles I and II of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). AB - A Joint Publication of The Walsh Center for Rural Health Analysis, National Opinion Research Center, University of Chicago, W Series, No. 6 and the RUPRI Center for Rural Health Policy Analysis, University of Nebraska Medical Center, P2004-6. PMID- 15529474 TI - The provider system for children's mental health: workforce capacity and effective treatment. AB - This issue brief examines two issues that are key to meeting children's unmet needs for mental health care: ensuring that the provider supply is adequate and that the care delivered is effective. It describes the shortage of qualified providers to address children's mental disorders, as well its possible causes; it describes how managed care, to a certain extent, drives practice patterns; and it discusses the gray areas in deciding which providers are most qualified to deliver what care. In addition, this paper introduces what is known about evidence-based care in children's mental health, the extent to which it is being taught and practiced, the extent to which health plans are adopting such practices, and the effect such strategies may have on the makeup of the children's mental health provider field. PMID- 15529475 TI - Predictors of depressive mood, occupational stress, and propensity to leave in older and younger mental health case managers. AB - Age differences and relationships between occupational stressors, work environment, depressed mood, and propensity to leave were examined among 263 case managers (CMs) in community-based mental health agencies. For younger CMs, satisfaction with the supervisor, perceived job pressure and lack of organizational support were significant predictors of propensity to leave the job. Older CMs who reported lower satisfaction with financial rewards, the type of work they did, and greater use of avoidance/resignation and help seeking coping strategies, were more likely to rate themselves higher on the depressed mood scale. PMID- 15529476 TI - A positive aspect of caregiving: the influence of social support on caregiving gains for family members of relatives with schizophrenia. AB - This study examines family members' caregiving gains as a result of caring for their relatives with schizophrenic spectrum disorders, and the influence of formal and informal social support on these positive experiences. The results from interviews with 560 family members support that the experience of caregiving gains is prevalent. Moreover, formal support from mental health professionals through information sharing and collaborative interactions with family members, and informal support from other family members, support group participation, and contributions from the relative with mental illness all have significant, positive associations with family members' experiences of caregiving gains. PMID- 15529477 TI - The nature and dimensions of social support among individuals with severe mental illnesses. AB - Recent research suggests that social support is associated with recovery from chronic diseases, greater life satisfaction, and enhanced ability to cope with life stressors. To further research in the area of social support and serious psychiatric disabilities, more reliable and valid measures are needed to assess this construct. The purpose of this study was to assess the psychometric properties of a widely used measure of social support (the Interpersonal Support Evaluation Checklist) among people with severe mental illness. We collected data on the ISEL's relationship to quality of life, self-esteem, psychiatric symptoms and vocational status among 147 participants. Factor and reliability analyses, as well as correlational analyses were undertaken. We found evidence for the reliability and validity of the ISEL when used with persons with severe mental illness. Taken together, our findings suggested that self-esteem, quality of life, and psychiatric symptoms were able to predict 38% of the variance in perceived social support. More favorable social supports increased the odds of being employed at 9 months into the study and social support was predictive of experiencing fewer psychiatric symptoms. Some forms of social support were perceived less favorably with age, but no other demographic or clinical variables significantly predicted perceived social supports. PMID- 15529478 TI - Social networks of people with dual diagnosis: the quantity and quality of relationships at different stages of substance use treatment. AB - This study compares the social networks, perceived levels of social support, and satisfaction with this support, for people with serious mental illnesses and comorbid problematic use of alcohol and other drugs who are at different stages of substance use recovery. Participants in the late stage of recovery were more likely to perceive greater social support from their non substance-using social contacts than participants in the early stage. This social support was more likely to be from professionals than other non-substance-using contacts in their networks. The support from professionals is discussed in relation to engagement in integrated dual diagnosis interventions. PMID- 15529479 TI - The importance of work as compared to other forms of daily occupations for wellbeing and functioning among persons with long-term mental illness. AB - This study investigated the importance of competitive work as compared to participating in community-based activities and having no regular daily activity for people with long-term mental illness. One hundred and three persons visiting an outpatient unit and representing the three targeted occupational conditions were randomized to the study and assessed on satisfaction with daily occupations, health, and wellbeing. Persons with work were more satisfied with their daily occupations and were interviewer-rated as better functioning than the other groups, but the groups did not differ on self-ratings of health and wellbeing. Unexpectedly, people participating in community-based activities did not show better satisfaction or functioning than those without regular daily activity. It was concluded that competitive work should be an urgent end goal for services designed for occupational or vocational support. PMID- 15529480 TI - Demographic analysis of members of the American Association of Community Psychiatrists. AB - This study examines which issues public psychiatrists perceive as having most impacted their careers over the past five years and whether this impact was experienced differently by psychiatrists in varying regions of the country. The author sent a questionnaire to all members of the American Association of Community Psychiatrists in clinical practice for at least five years. Seventy-one percent (344 of 482) responded. As expected, respondents were generally negative about how economic factors have impacted their professional lives. This impact, however, was experienced very differently depending on which region of the country the psychiatrist was working. PMID- 15529481 TI - Public psychiatrists' perceptions of changes over the past five years. AB - This study examines which issues public psychiatrists perceive as having most impacted their careers over the past five years and whether this impact was differentially experienced by staff psychiatrists and medical directors. The authors sent a questionnaire to all members of the American Association of Community Psychiatrists (AACP) in clinical practice for at least 5 years. Surprisingly, public psychiatrists report that the impact of recent changes have been primarily positive, except in the domain of economic developments. Both program and agency medical directors report experiencing about the same amount of change, though significantly more positive overall impact, compared to staff psychiatrists. This finding should encourage psychiatrists to become program medical directors, a feasible next step for many staff psychiatrists. PMID- 15529483 TI - Which adults with acute diarrhea should be evaluated? What is the best diagnostic approach? PMID- 15529484 TI - Dementia with Lewy bodies: diagnosis and clinical approach. AB - Dementia with Lewy bodies (DLB) is a common form of dementia that is being increasingly recognized. This paper reviews this disorder, emphasizing how it is different from other causes of dementia. PMID- 15529485 TI - Hospital management of diabetes: beyond the sliding scale. AB - A growing body of evidence suggests that there is a close correlation between good glucose control and improved clinical outcomes in hospitalized diabetic patients. Until recently, however, no established standards have existed for caring for people with diabetes in the hospital. At a recent consensus conference, experts from around the world studied the evidence and recommended new glycemic targets for hospitalized patients (Endocr Pract 2004; 10:77-82). The challenge now is to implement these targets. PMID- 15529486 TI - A construction worker with recent confusion, disorientation, and somnolence. PMID- 15529487 TI - Drug-eluting stents: the beginning of the end of restenosis? AB - In multiple clinical trials, patients who received drug-eluting stents instead of plain stents during percutaneous coronary interventions had rates of restenosis that were lower by roughly one half to three fourths, depending on how restenosis was defined and on the population studied. These stents will likely be used more and more as their indications evolve. PMID- 15529488 TI - Drug-eluting stents are here--now what? Implications for clinical practice and health care costs. PMID- 15529489 TI - Treating osteoporosis in post-menopausal women: a case approach. AB - We now have several agents of different classes for treating postmenopausal osteoporosis. In this paper, a case report serves as the focus for a discussion of the risk factors for postmenopausal osteoporosis and of the available therapies. PMID- 15529490 TI - Two interceptive approaches to palatally displaced canines: a prospective longitudinal study. AB - This study evaluated the effectiveness of two interceptive approaches to palatally displaced canines (PDCs), ie, extraction of the deciduous canines alone and in association with the use of a cervical pull headgear. The prospective longitudinal design of the investigation included 46 subjects with PDC (62 maxillary canines) who were randomly assigned to one of three groups (1) a group that underwent the extraction of the deciduous canine only, (2) a group that received in addition the use of a cervical pull headgear, and (3) an untreated control group. Panoramic radiographs were evaluated at initial observation (T1) and after an average period of 18 months (T2). Cervical vertebral maturation was assessed on lateral cephalograms at T1. Successful or unsuccessful canine eruption was assessed 48 months after T1. The between-group statistical comparisons were performed on the T1-T2 changes in the diagnostic parameters on panoramic radiographs, the prevalence rates of successful canine eruption, and the amount of time for canine eruption. The removal of the deciduous canine as an isolated measure to intercept palatal displacement of maxillary canines showed a prevalence rate of 50% success, which was not significantly greater than the success rate in untreated controls. The use of a headgear in addition to the extraction of the deciduous canine induced successful eruption in 80% of the cases, with a significant improvement in the measures for intraosseous canine position. There was no significant difference between the two interceptive approaches in the time required for canine eruption. PMID- 15529491 TI - Three-dimensional facial imaging: accuracy and considerations for clinical applications in orthodontics. AB - This study determined the accuracy of a camera system capable of recording three dimensional facial images. A Rainbow 3D Camera Model 250 system (Genex Technologies Inc, Kensington, Md) was used to capture images of specific models: (1) a precalibrated precision model and (2) a mannequin model that served to simulate the human condition. To assess the accuracy of the camera system, repeated images of both models were recorded at two time points, one week apart. Repeated measurements of specific distances were recorded directly on the models and from each image. Means and standard deviations were calculated for all the repeated measurements at each time point. A two-tailed t-test was used to test for significant differences between (1) each distance measured directly on the precision model and the same distance measured on the images of the precision model, (2) each distance measured directly on the mannequin and the same distance measured on the images of the mannequin, and (3) the mean differences between the same distances measured at the two times. The findings showed that substantial image distortion occurred when images of sharp angles (90 degrees) were captured. Also, those images captured from the frontal perspective +/- 15 degrees were the most accurate. PMID- 15529492 TI - Stability of Class II, division 1 treatment with the headgear-activator combination followed by the edgewise appliance. AB - This study assessed the stability of the headgear-activator combination treatment, followed by edgewise mechanotherapy, 5.75 years after treatment. The experimental group consisted of 23 patients who were evaluated during treatment and after treatment. Two compatible control groups consisting of 15 Class II, division 1 patients and 24 normal occlusion individuals were used. This enabled us to evaluate the changes during treatment and after treatment, respectively. Results showed that the anteroposterior dentoalveolar changes and the maxillary and the mandibular positions remained stable in the long term. However, there was a slight relapse of the maxillomandibular relationship probably because the maxilla resumed its normal development and the mandibular growth rate was smaller than in the control group. The overbite demonstrated a statistically significant relapse that was directly proportional to the amount of its correction. There were low but significant inverse correlations between the changes in Go-Gn during and after treatment. These included the uprighting of the maxillary incisors, labial tipping of the mandibular incisors, and the amount of molar relationship correction during treatment and their stability. Active retention time, length of posttreatment period, initial Class II malocclusion severity (ANB and Wits), and initial molar relationship did not present any correlation with molar relationship and overjet relapse. However, the initial overjet presented a low but statistically significant correlation with molar relationship relapse and overjet relapse. PMID- 15529493 TI - The effects of early preorthodontic trainer treatment on Class II, division 1 patients. AB - The aim of this study was to clarify the dentoskeletal treatment effects induced by a preorthodontic trainer appliance treatment on Class II, division 1 cases. Twenty patients (10 girls and 10 boys, mean age 9.6 +/- 1.3 years) with a Class II, division 1 malocclusion were treated with preorthodontic trainer appliances (Myofunctional Research Co., Queensland, Australia). The patients were instructed to use the trainer every day for one hour and overnight while they slept. A control group of 20 patients (mean age 10.2 +/- 0.8 years) with untreated Class II, division 1 malocclusions was used to eliminate possible growth effects. Lateral cephalograms were taken at the start and end of treatment. Final cephalograms were taken 13.1 +/- 1.8 months after trainer application, compared with a mean of 11.2 +/- 2.4 months later for the control group. The mean and standard deviations for cephalometric measurements were analyzed by paired samples t-test and independent-samples t-tests. At the end of the study period, the trainer group subjects showed significant changes including anterior rotation and sagittal growth of the mandible, increased SNB and facial height, reduced ANB, increased lower incisor proclination, retroclination of upper incisors, and overjet reduction. However, only total facial height increase, lower incisor proclination, and overjet reduction were significantly higher when compared with the changes observed in the control group. This study demonstrates that the preorthodontic trainer application induces basically dentoalveolar changes that result in significant reduction of overjet and can be used with appropriate patient selection. PMID- 15529494 TI - Titanium screw implants in optimization of radiographic evaluation of facial growth in longitudinal animal studies. AB - The aim of this study was to develop a method that optimizes the reliability of longitudinal radiographic evaluation of small and fast-growing animals, such as the rabbit. Because the use of conventional cephalometric methods, including superimposition of serial radiographs, is more problematic in small animals than in humans, two titanium-alloy screws were placed 10 mm apart in the sagittal crest of the parietal bone in 10 growing New Zealand white rabbits. The anterior screw served as holder for a steel pin that, in turn, secured the fixation of the rabbit's head to a specially designed cephalostat. A lateral cephalogram of each animal was exposed on four occasions at one-month intervals. Computer-aided superimpositions were made of all four cephalograms from each animal using the screws in the calvarium as reference structures. To evaluate the method, the superimpositions were repeated after three to eight weeks, and the superimposition reproducibility was calculated. From the results, it can be concluded that the method allows congruent positioning of the animal skull relative to the film-focus assembly at repeated radiographic examinations. Furthermore, it introduces readily identified reference structures in the animal skull that can be used at high-precision superimposition of serial radiographs. PMID- 15529495 TI - Changes in dental arch measurements of young adults with normal occlusion--a longitudinal study. AB - This study evaluates the longitudinal changes in occlusal dimensions in young adults. The sample was composed of 27 individuals (13 male and 14 female individuals) who presented dental Class I and normal occlusion, according to the inclusion protocol. All teeth were present except for the third molars. The mean age of the subjects was 21 years and two months initially and 28 years and four months at the end of the observation period. The measurements collected in both arches were overbite, overjet, intercanine and intermolar distances, irregularity index, and perimeter. All measurements were taken from study dental casts by a calibrated operator, using a digital caliper. The paired t-test was used to evaluate the changes, using the difference between the initial and final mean measurements. Data indicated that the overbite increased 0.39 mm, the incisor irregularity increased 0.38 mm in the upper arch and 0.54 mm in the lower arch, and the arch perimeter decreased 0.67 mm in the upper arch and 0.71 mm in the lower arch (P > .05). The other measurements did not show significant changes. No sexual dimorphism was observed. It could be concluded that occlusal dimensions change throughout adult life. In the sample studied, changes were statistically significant for overbite, incisors irregularity, and arch perimeter after a mean period of seven years and two months. These changes can be observed in both sexes. PMID- 15529496 TI - Longitudinal posteroanterior changes in transverse and vertical craniofacial structures between 10 and 14 years of age. AB - Longitudinal growth changes in the transverse and vertical craniofacial dimensions occurring between 10 and 14 years of age were evaluated in 22 female and 23 male subjects. All subjects were healthy, had a clinically acceptable occlusion, and had not received orthodontic or orthopedic therapy before. Serial cephalometric radiographs were taken at ages 10, 11, 12, and 14 years. In studying the P-A cephalograms, seven transverse and five vertical measurements were used. The effects of age and sex on the transverse and vertical growth of the craniofacial structures were investigated by means of analysis of variance and Bonferroni multiple comparison test. The results of the study revealed that all measurements studied were affected by age, and cranial, facial, nasal, and maxillary widths were affected by sex. The most pronounced age-related increases occurred in the mandibular width for transverse measurements and in the total facial height for vertical measurements in both sexes during this study. PMID- 15529497 TI - A new method of predicting mandibular length increment on the basis of cervical vertebrae. AB - The purpose of this study was to establish an equation to predict incremental mandibular length on the basis of the analysis of the cervical vertebrae in a single cephalometric radiograph and to compare the predictive accuracy with other methods. Data comprised two groups of 23 Japanese girls between CVMS I and CVMS V. Group A was examined to construct the predication equation. Group B served to compare the predictive accuracy with the growth potential method and the growth percentage method. The following results were obtained: (1) an equation was determined to obtain mandibular length increments on the basis of the measurements in the third and fourth cervical vertebral bodies, and (2) the average error between the predicted increment and the actual increment was 1.5 mm for the equation method, 2.4 mm for the growth potential method, and 2.8 mm for the growth percentage method. These results suggest that with the use of cervical vertebral measurements, it may be possible to evaluate the mandibular growth potential. PMID- 15529498 TI - Malocclusion and crowding in an orthodontically referred Turkish population. AB - This study was conducted to evaluate malocclusion and crowding in 1356 patients (793 girls, 563 boys) referred to the Department of Orthodontics, Suleyman Demirel University, Turkey. Class I was the most frequently seen malocclusion in this referred Turkish orthodontic population whereas Class II, division 2 was the least frequently seen. Comparison of mean ages of the malocclusion groups indicated statistically significant difference between Class I and Class II, division 1 groups (P < .05). The lowest mean age was present in the Class II, division 1 group. Mild mandibular crowding was the most common finding whereas severe mandibular crowding was seen least frequently in all malocclusion groups. Cross tabulation of maxillary and mandibular crowding indicated that mild maxillary and severe mandibular crowding in the same patient was rarely seen in all types of malocclusions. Moderate maxillary and severe mandibular crowding in the same patient was another rare finding for all malocclusion groups. PMID- 15529499 TI - Facial profile preferences among various layers of Turkish population. AB - There are many criteria and methods used to constitute and determine a standardized esthetic concept in orthodontics. However, the subjectivity of the esthetic concept is the common opinion of the authors. Ethnic and racial differences play a major role in diversifying esthetic preferences. The aims of this study were to determine the general esthetic preference of a Turkish population and to find out whether this preference was affected by sex, age, education, social status, geographic location, or personal profile. Toward these aims, eight profile estimates for each sex were morphed by a video imaging technique and then scored by 400 participants. Analysis of variance (ANOVA) and independent sample t-tests were used to compare the preferences of the groups. The orthognathic profile in both sexes was selected as the most preferred profile whereas the convex profile with a prognathic maxilla and a retrognathic mandible were the least preferred. The public also admired fuller and protrusive lips in females and retrusive lips with a prominent nose and chin in males. Sex, age, education, social status, geographic location, and personal profile were also shown to affect the public's profile preferences. PMID- 15529500 TI - Maxillary morphology in obstructive sleep apnea: a cephalometric and model study. AB - The relationship between maxillary constriction and the etiology of obstructive sleep apnea (OSA) is not clear. This prospective case-control study compared maxillary morphology in 94 dentate subjects (47 OSA and 47 control subjects), using upright lateral cephalograms and study models. Each subject had height, weight, and neck circumference measurements recorded and underwent an orthodontic examination. An upright lateral cephalogram and dental impressions were obtained. All data were analyzed using the SPSS statistical package applying nonparametric tests at the 5% level of significance. Male and female subjects were examined separately, and statistically significant differences were found between the cephalometric measurements for OSA and the control subjects. The palatal angle was more obtuse in male OSA subjects (P < .05). The PNS-posterior pharyngeal wall was shorter (P < .05) and the soft palate longer in female OSA subjects (P < .05). Minimum palatal airway widths were significantly reduced in both male (P < .01) and female (P < .001) subjects. In the comparison of study model measurements, palatal heights in OSA subjects were greater (P < .05). Thus, maxillary morphological differences do exist between OSA and control subjects, supporting their role as a etiological factor. PMID- 15529501 TI - Relationships between dental and skeletal maturity in Turkish subjects. AB - The aim of this study was to investigate the relationships between the stages of calcification of various teeth and skeletal maturity stages among Turkish subjects. The samples were derived from dental panoramic and hand-wrist radiographs of 500 subjects (215 males and 285 females). Calcification of the mandibular canines, first and second premolars, and second and third molars was rated according to the system of Demirjian. To evaluate the stage of skeletal maturation of each hand-wrist radiograph, nine ossification events were determined according to the systems of Bjork, and Grave and Brown. Statistically significant relationships were determined between dental calcification and skeletal maturity stages according to Spearman rank-order correlation coefficients. Correlations between dental development and skeletal maturity ranged from .490 to 0.826 for females and .414 to .706 for males (P < .01). The second molar showed the highest correlation and the third molar showed the lowest correlation for female and male subjects. For both sexes, root formation of the canine as well as the first premolar was completed in the majority of the subjects at the MP3cap, PP1cap, Rcap stages. Because of the high correlation coefficients, this study suggests that tooth calcification stages from panoramic radiographs might be clinically useful as a maturity indicator of the pubertal growth period. It is appropriate to put these skeletal and dental maturation relationships into daily orthodontic diagnostic practice, when treating a Turkish patient. PMID- 15529502 TI - Absence of association between transforming growth factor-beta1 promoter polymorphisms and hypodontia. AB - Hypodontia, the congenital absence of one or a few teeth, is one of the most common alterations of the human dentition. The most common permanent missing teeth are the third molars, second premolars, and maxillary lateral incisors. Although hypodontia does not represent a serious public health problem, it may cause masticatory and speech dysfunctions and esthetic problems. Transforming growth factor-beta1 (TGF-beta1) is believed to play an important role in tooth development. Its gene is expressed at bud, cap, and bell stages of odontogenesis. Genetic polymorphisms in the TGF-beta1 gene promoter were shown to interfere with the transcriptional activity of this gene. To further investigate the role of the TGF-beta1 gene in human hypodontia, we analyzed the frequencies of the -509 polymorphism (C-T) alleles and -800 polymorphism (G-A) alleles and genotypes in the TGF-beta1 gene promoter in 51 Caucasian subjects with hypodontia and 48 control individuals. Our data suggest that these TGF-beta1 promoter polymorphisms are not associated with hypodontia. PMID- 15529503 TI - The relationship between bruxism and occlusal factors among seven- to 19-year-old Turkish children. AB - The aim of this study was (1) to investigate the relationship between occlusal factors and bruxism among 965 Turkish subjects (472 boys and 493 girls) with a mean age of 12.8 years (range, seven to 19 years); and (2) to identify possible sex differences between girls and boys. This sample was divided into two groups of bruxers or nonbruxers based on a clinical examination and self-reports. The examiner recorded the Angle molar classification bilaterally, severity of anterior crowding, existence of anterior and posterior crossbite, open and deep bite, functional shift, and excessive overjet. The relationships between occlusal factors and bruxism and sex differences between boys and girls were evaluated with chi-square analysis (chi2) using the SPSS software package. The results showed that bruxism was diagnosed in 12.6% of all subjects. The evaluation of the findings indicated that no statistically significant relationships were determined between bruxism and occlusal factors. No sex differences were found between occlusal factors in relation to bruxism. The prevalence of bruxism in boys and girls was similar, and no statistically significant differences were found. It is concluded that none of the occlusal factors seem to play a role in the development of bruxism. However, additional longitudinal studies with larger samples need to be conducted to determine if there is any relationship between occlusal factors and bruxism. PMID- 15529504 TI - Alterations of the rat temporomandibular joint in functional posterior displacement of the mandible. AB - Functional malocclusion that induces posterior condylar displacement may affect the remodeling processes of the temporomandibular joint structures. We tested the hypothesis that intermittent posterior condylar displacement due to functional malocclusion traumatizes condylar cartilage and joint innervated nerve fibers. Thirty-nine eight-week-old Wistar rats were used. To induce functional posterior condylar displacement, guiding appliances were attached to maxillary incisors of 24 rats for four, seven, and 14 days. Fifteen normal rats served as controls. Sections were stained with hematoxylin and eosin or processed for immunohistochemistry of protein gene product 9.5 and growth-associated protein-43 (GAP-43). Functional posterior condylar displacement led to a diminution in proliferative cells, reduction in cartilage width, and re-expression of GAP-43 immunoreactive nerve fibers. These results indicate that intermittent posterior condylar displacement due to functional malocclusion causes dysfunctional remodeling of condylar cartilage and nerve injury. PMID- 15529505 TI - Comparison of skeletal and dental morphology in asymptomatic volunteers and symptomatic patients with bilateral disk displacement without reduction. AB - The purpose of this study was to evaluate the effect of bilateral disk displacement without reduction (BDDN) on the skeletal and dental pattern of affected individuals. There were 59 symptomatic female patients and 46 asymptomatic normal female volunteers. All study participants had bilateral high resolution magnetic resonance imaging scans in the sagittal (closed and open) and coronal (closed) planes to evaluate the temporomandibular joints. Linear and angular cephalometric measurements were taken to evaluate the skeletal, denture base, and dental characteristics of the two groups. A smaller cranial base length (Ba-Na) was found in the symptomatic group. The facial plane angle was smaller, and the angle of convexity was larger because of the retropositioned mandible. The lower denture base was also retruded as shown by the smaller SNB angle. The BDDN group exhibited a larger overjet. The mandibular plane angle was steeper, the Y-axis was more vertical (S-Gn to FH), the posterior ramal height (Ar-Go) was shorter, and the angle between the mandibular and the palatal plane (PP to MP angle) was increased in the symptomatic group. No significant dental differences were found. This study showed that alterations in skeletal morphology might be associated with BDDN. This study suggests that subjects with BDDN may manifest altered craniofacial morphology. The clinician should be aware of that possibility, especially for the growing patients and the surgical candidates. PMID- 15529506 TI - Stiffness in bending of a superelastic Ni-Ti orthodontic wire as a function of cross-sectional dimension. AB - Superelasticity is a property used in orthodontics to initiate tooth movement in the first stage of orthodontic treatment. It is the aim of all clinicians to accomplish biological tooth movement, which implies the use of low, continuous force and requires archwire with low stiffness. In this study, 15 nickel-titanium archwires with three different cross-sectional dimensions were tested in three point bending to determine the nature of forces in a loading and unloading cycle. The evolution of stiffness in bending as a function of wire size is discussed. The applied forces or stiffness dependence on cross-sectional size differs from the linear-elastic prediction because of the superelasticity property. We discuss the origin of the nonconventional profile of curves and the nature of reversible large deformation of these alloys. Martensitic transformation is at the origin of nonlinear elasticity. The stiffness decreases with increasing deflection, and this phenomenon is emphasized in the unloading process. The value of stiffness appears to vary with wire size but depends on the ratio of volume of martensitic transformation. During martensitic transformation, the rigidity (elastic modulus) of the alloy is nonconstant. These results and their understanding should allow a different approach of biomechanical considerations, ie, a large-size square wire does not produce necessarily high forces. PMID- 15529507 TI - Are the flowable composites suitable for orthodontic bracket bonding? AB - The study aims to determine the shear bond strength (SBS) values of different flowable composites (Pulpdent Flows-Rite, 3M Filtek Flow, and Heraeus Kulzer Flow Line) in comparison with a conventional orthodontic adhesive and the bond failure sites of these composites. Eighty extracted human premolars were divided into four groups of 20 teeth each. Brackets were bonded to the teeth in each test group with different composites, according to the manufacturer's instructions. SBS values of these brackets were recorded (in MPa) using a universal testing machine. Adhesive remnant index (ARI) scores were determined after the failure of brackets. Data were analyzed using analysis of variance (ANOVA), Tukey honestly significant difference, and chi-square tests. SBS values of groups 1 (Transbond XT), 2 (Flows-Rite), 3 (Flow), and 4 (Flow Line) were found to be 17.10 +/- 2.48 MPa, 6.60 +/- 3.2 MPa, 7.75 +/- 2.9 MPa, and 8.53 +/- 3.50 MPa, respectively. The results of this study demonstrate that the orthodontic adhesive (Transbond XT) had higher SBS values than the flowable composites. Results of ANOVA revealed statistically significant differences among the groups (P < .05). The SBS values were significantly lower in all flowable composite groups than the orthodontic adhesive. ARI scores were significantly different between the orthodontic adhesive and all the flowable groups investigated. The use of flowable composites is not advocated for orthodontic bracket bonding because of significantly lower SBS values achieved. PMID- 15529508 TI - Sliding mechanics with microscrew implant anchorage. AB - Three cases are illustrated. One was treated with maxillary microscrew implants, another with mandibular microscrew implants, and the third with both maxillary and mandibular microscrew implants. With the maxillary microscrew implants, the maxillary anterior teeth were retracted bodily with a slight intrusion and all the premolar extraction space was closed without loss of anchorage. Furthermore, the maxillary posterior teeth showed distal movement. The mandibular microscrew implants controlled the vertical position of the mandibular posterior teeth and played an important role in improving the facial profile. The efficacy of sliding mechanics with microscrew implant anchorage on the treatment of skeletal Class II malocclusion is also discussed. PMID- 15529509 TI - The efficient use of midpalatal miniscrew implants. AB - The midpalatal suture area with thin, keratinized soft tissue and sufficient cortical bone is an optimal site for miniscrew implantation. Even though the midpalatal area has its own anatomical limitations, it can be overcome by bonding extension arms to the miniscrews. This technique also enhances screw stability, thus producing a successful outcome. Among many applications, midpalatal miniscrew implants can be effectively used for intrusion of upper molars with reduced treatment time and enhanced patient comfort. PMID- 15529510 TI - Screw-type implants used as anchorage for lingual orthodontic mechanics: a case of bimaxillary protrusion with second premolar extraction. AB - We present a case of bialveolar protrusion treated with second premolar extraction. The patient did not agree to placement of a visible labial appliance or to the use of a headgear. Therefore, a lingual orthodontic appliance was used, and titanium screws were placed into the buccal alveolar bone for orthodontic absolute anchorage and support of en masse retraction of the anterior teeth. Cephalometric superimposition and panoramic radiographs showed little anchorage loss and good occlusion at the end of treatment. Our results suggest that lingual treatment combined with a screw-type implant anchorage provides reliable and comfortable results for those seeking invisible treatment. PMID- 15529511 TI - Re: Gidarakou IK, et al. Comparison of skeletal and dental morphology in asymptomatic volunteers and symptomatic patients with bilateral degenerative joint disease. Angle Orthod. 2003;73:71-78. PMID- 15529512 TI - Diabetic and hypertension guidelines updated. AB - The guidelines are clear. We need optimal control of blood pressure and lipid profiles. Diabetics need particular aggressive risk reduction. No risk factor should be managed in isolation--overall risk reduction is essential. We have a job to do and must do it thoroughly--we now need a programme of early detection and that needs political will and investment. PMID- 15529513 TI - Raising high-density lipoprotein. PMID- 15529514 TI - Durable efficacy of metformin/glibenclamide combination tablets (Glucovance) during 52 weeks of open-label treatment in type 2 diabetic patients with hyperglycaemia despite previous sulphonylurea monotherapy. AB - Oral anti-diabetic combinations that address insulin resistance and beta-cell dysfunction (e.g. metformin and glibenclamide) represent a rational therapeutic option for patients uncontrolled on monotherapy. A 52-week, open-label extension to a double-blind study evaluated metformin-glibenclamide combination tablets (Glucovance) in 477 patients with hyperglycaemia despite sulphonylurea therapy. Reductions in HbA1C were maintained, with a mean reduction of -1.7% after 52 weeks, compared with the baseline value for the double-blind trial. Eighty-five patients receiving 4 x 500 mg/2.5 mg tablets daily displayed a marked improvement in HbA1c following up-titration to a regimen of 2 x 500 mg/2.5 mg + 3 x 500 mg/5 mg tablets. Lipid profiles improved significantly. The combination tablets were well tolerated: 11.1% of patients reported hypoglycaemic symptoms (all either mild or moderate severity). No patient withdrew or required pharmacologic intervention for hypoglycaemia. Metformin-glibenclamide combination tablets are an effective and well-tolerated therapeutic option for intensifying oral anti diabetic therapy. PMID- 15529515 TI - Effectiveness and tolerability of a homoeopathic remedy compared with conventional therapy for mild viral infections. AB - Treatments for mild viral infections are usually directed at providing symptomatic relief. The effectiveness of the homoeopathic remedy Gripp-Heel was compared with that of conventional treatments in a prospective, observational cohort study in 485 patients with mild viral infections and symptoms such as fever, headache, muscle pain, cough or sore throat. Practitioners specialised in homoeopathy or conventional treatment, or practised both to similar extents. As evaluated by the practitioners, the homoeopathic therapy was effective to similar or greater degree than the conventional therapies: 67.9% of patients were considered asymptomatic at the end of Gripp-Heel therapy vs. 47.9% of patients in the control group. Practitioners judged homoeopathic treatments as 'successful' in 78.1% of cases vs. 52.2% for conventional therapies. Tolerability and compliance were good in both treatment groups, with the verdict 'very good' given for 88.9% of patients in the homoeopathic group vs. 38.8% in the conventional treatment group. PMID- 15529516 TI - Cardiovascular effects of treatment of type 2 diabetes with pioglitazone, metformin and gliclazide. AB - Cardiovascular mortality and morbidity are increased in patients with type 2 diabetes. However, there are few data from clinical trials comparing cardiovascular effects of alternative oral anti-diabetic agents. Major cardiovascular outcomes during four one-year, double-blind trials in over 3700 patients with type 2 diabetes randomised to either a thiazolidinedione, pioglitazone, metformin or a sulphonylurea, gliclazide treatment have been combined. Mean blood pressure was slightly reduced by all treatments, with pioglitazone treatment resulting in the largest falls (approximately 1.5 mmHg). Hospitalisations for cardiac or cerebrovascular events were similar with the different treatments. Overall mortality was seven of 1857 for pioglitazone and 10 of 1856 for non-pioglitazone treatments, of which three and six were cardiac deaths, respectively. The incidence of congestive cardiac failure was similar with pioglitazone (12/1857) and non-pioglitazone (10/1856) treatments. The results show similar cardiovascular outcome for the three different treatments over a one-year period, but demonstrate interesting differences, which will require longer-term formal outcome studies to determine their significance. PMID- 15529517 TI - Frequency of angiotensin-converting enzyme gene polymorphism in Turkish hypertensive patients. AB - Hypertension is a multifactorial disease, in which genetic factors play an important role. This study was carried out to determine angiotensin-converting enzyme levels and angiotensin-converting enzyme gene polymorphism in Turkish hypertensive patients, and to establish whether there is an association of angiotensin-converting enzyme gene polymorphism with clinical and echocardiographic parameters. We have investigated the association among the allelic distribution of the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme gene identified by polymerase chain reaction, angiotensin-converting enzyme activity determined spectrophotometrically, cardiac morphology and function assessed by means of echocardiography. Distribution of angiotensin-converting enzyme gene I/D polymorphism and allele frequencies in hypertensive patients was not significantly different from controls. D allele frequency was 51.7% in hypertensives vs. 51.9% in controls and I allele 48.3 vs. 48.1%, respectively. The level of angiotensin-converting enzyme activity was significantly higher in the patients homozygotes for D allele (DD = 59.93 U/l) than in heterozygotes (ID = 39.49) and in homozygotes for I allele (II = 40.28 U/l). In addition to these, the level of angiotensin-converting enzyme activity was significantly lower in the ID and especially II patients receiving ACE inhibitors than the others. Also, it was determined that left atrium diameter was larger in the patients homozygotes for I allele than the others. PMID- 15529518 TI - The case for blood pressure control in risk groups. AB - Hypertension rarely occurs in isolation, and many hypertensives have additional risk factors for cardiovascular disease in addition to elevated blood pressure. Each patient's cardiovascular disease risk profile should be determined individually, and the treatment approach tailored to each case. Cardiovascular disease risk factors and high blood pressure are closely linked, suggesting that the ideal treatment should not only lower blood pressure, but also effectively lower overall risk. This is likely to require more than one drug, and one of the most effective and safe combinations is that of an angiotensin receptor blocker with a diuretic. The completion of one of the most important trials undertaken to explore risk factors and anti-hypertensive treatment, the Valsartan long-term evaluation trial (VALUE), will certainly enhance understanding for the role of combination treatment in high-risk patients, as well as contribute to the design of rational treatments for blood pressure control. PMID- 15529519 TI - Imaging in psychiatric illnesses. AB - Brain imaging has given new insights into the structure and function of the brain in psychiatric illnesses. The conditions studied include Alzheimer's disease, Schizophrenia, depression and psychopathic disorders. Emerging technologies in the field of brain imaging have helped in the understanding of pathophysiology, aetiology, diagnosis, treatment response and prognosis of certain psychiatric disorders. This article summarizes the literature available and the potential clinical applications. PMID- 15529520 TI - Cholesterol metabolism and its implications for therapeutic interventions in patients with hypercholesterolaemia. AB - Cardiovascular diseases are the principal causes of mortality in middle-aged people and in older people. Coronary heart disease (CHD) is the most common of the cardiovascular diseases; high serum levels of cholesterol are associated with atherosclerosis and an increased risk of CHD. Cholesterol homeostasis is achieved by means of a fine balance between cholesterol intake, absorption/excretion and synthesis. All of these processes are tightly linked and a change in one of them can significantly influence the others. Results from both experimental studies and clinical trials have shown that inhibition of cholesterol synthesis with a statin increases absorption and that conversely, inhibition of cholesterol absorption increases synthesis. The tight linkage of cholesterol absorption and synthesis in maintaining cholesterol homeostasis suggests that treatment with an agent that influences only one of these two processes is likely to have distinct limits with respect to its effects on cholesterol levels. Better understanding of cholesterol homeostasis, particularly the close interrelationship between cholesterol synthesis and absorption, may result in the design of rational integrated treatment regimens that employ multiple agents with complementary actions that attack multiple mechanisms to lower cholesterol. PMID- 15529521 TI - Avandamet: combined metformin-rosiglitazone treatment for insulin resistance in type 2 diabetes. AB - Insulin resistance is a major endocrinopathy underlying the development of hyperglycaemia and cardiovascular disease in type 2 diabetes. Metformin (a biguanide) and rosiglitazone (a thiazolidinedione) counter insulin resistance, acting by different cellular mechanisms. The two agents can be used in combination to achieve additive glucose-lowering efficacy in the treatment of type 2 diabetes, without stimulating insulin secretion and without causing hypoglycaemia. Both agents also reduce a range of atherothrombotic factors and markers, indicating a lower cardiovascular risk. Early intervention with metformin is already known to reduce myocardial infarction and increase survival in overweight type 2 patients. Recently, a single-tablet combination of metformin and rosiglitazone, Avandamet, has become available. Avandamet is suitable for type 2 diabetic patients who are inadequately controlled by monotherapy with metformin or rosiglitazone. Patients already receiving separate tablets of metformin and rosiglitazone may switch to the single-tablet combination for convenience. Also, early introduction of the combination before maximal titration of one agent can reduce side effects. Use of Avandamet requires attention to the precautions for both metformin and rosiglitazone, especially renal, cardiac and hepatic competence. In summary, Avandamet is a single-tablet metformin rosiglitazone combination that doubly targets insulin resistance as therapy for hyperglycaemia and vascular risk in type 2 diabetes. PMID- 15529522 TI - Adefovir dipivoxil: review of a novel acyclic nucleoside analogue. AB - Adefovir dipivoxil (ADF) is a novel acyclic nucleoside analogue that has recently been approved for the treatment of chronic hepatitis B virus (HBV). Adefovir was initially assessed at higher doses for the treatment of human immunodeficiency virus (HIV) infection. However, in these studies, nephrotoxicity proved a dose limiting side effect. Large randomised controlled studies have recently shown that ADF results in histological, virological and biochemical improvement in both hepatitis B e antigen (HBeAg)-positive and HBeAg-negative chronic HBV. While the rate of HBeAg seroconversion at 1 year (12%) was lower than both lamivudine and interferon, this increases with prolonged treatment. The clinical improvements occurred without serious side effects or the development of resistance at the dose of 10 mg daily, in treatment trials of up to 2 years, although resistance has now been observed. In addition, the drug is efficacious in HBV/HIV co infection and hepatitis B-infected liver transplant recipients, particularly in those who have developed lamivudine resistance. ADF can be added as a treatment option to existing treatment options (interferon-alpha and lamivudine) and assumes a role in the ongoing management of chronic HBV. The optimal use of ADF as either a monotherapy or as part of combination therapy requires further assessment. PMID- 15529523 TI - The place of the ice pack test in the diagnosis of myasthenia gravis. AB - In the evaluation of ptosis, the ice pack test is a simple, quick, bedside method to ascertain whether it is of myasthenic origin, which requires no specialised medication or equipment and is free of reported adverse effects. Pooling of published studies suggests very high sensitivity, specificity and likelihood ratios, but standardisation and operationalisation of the test are important. False-negative tests may occasionally occur. PMID- 15529524 TI - A case of condyloma acuminatum treated successfully with low-dose isotretinoin and interferon. AB - The treatment of condyloma acuminatum may be problematic in clinical practice. We present a 27-year-old man with condyloma acuminatum who was treated successfully with a combination therapy of isotretinoin and interferon-alfa-2a. In the previous reports of combination therapy, isotretinoin has been given in a dosage of 1 mg/kg/day. However, we achieved the same success at a dosage of 0.5 mg/kg/day of isotretinoin. PMID- 15529525 TI - Severe sepsis following wound infection by an unusual organism--Clostridium novyi. AB - We present a case of post-operative wound infection with Clostridium novyi in a non-intravenous drug user. Clinical features included progressive cellulitis despite being on antibiotics, accompanied by hypotension, marked leucocytosis and oedema but minimal fever. While established infection with this organism is associated with high mortality, our patient survived. The administration of clindamycin and intravenous immunoglobulin in addition to early surgical assessment and aggressive debridement of affected tissue may have contributed to this successful outcome. To our knowledge, this is the only reported post operative wound infection due to this pathogen. PMID- 15529526 TI - Pitfalls in the differential diagnosis of a pelvic cyst: lessons from a post menopausal woman with bladder diverticulum. AB - We present a case of non-obstructive bladder diverticulum in a 75-year-old post menopausal woman. An ovarian cyst was previously suspected, which resulted in a futile exploratory laparotomy without making any definite diagnosis, 1 year earlier. During this admission, transvaginal ultrasound-guided cyst aspiration was arranged to determine the nature of the presumed 'recurrent' cyst and to relieve the symptoms. Prior to cyst aspiration, up to 700 ml of urine through urinary catheterisation and the gradual disappearance of the 'cyst' alerted us to the possibility of a bladder diverticulum, which was later confirmed by retrograde cystography. This case illustrates the lessons that despite considerable researches having been done on enhancing sonographic accuracy, diagnosis based on imaging alone is likely to be associated with multiple pitfalls. Recognising the common pitfalls and integrating clinical information and alertness with ultrasonic features remains the mainstay of sonographic differential diagnosis. PMID- 15529527 TI - [Results of a multi-year use of combined laser-surgical treatment in tumors of the epibulbar and limbocorneal localizations]. AB - Fifty-five patients (55 eyes) were followed up. The "Romashka-2" CO2-laser plant was used for treatment of epithelial and epibulbar tumors (41 patients, 41 eyes). Microsurgical interventions with subsequent lamellar or penetrating keratoplasty were made in 14 patients (14 eyes). The experimental group of patients was followed up from 3 to 10 years. Relapses were registered in 4 eyes (4 patients). The CO2-laser treatment had a number of advantages before surgery or cryotreatment in removing the epibulbar tumors without growth in the cornea. Ablasticity, minimal trauma to tissues and evaporation of diffusive neoplasms can be named among such advantages. In cases of growth of epibulbar tumors in the cornea, there were direct indications for keratoplasty with preliminary barrage of CO2 irradiation on the sides of the mucous tunic of epibulbar tumor. Tumors were surgically removed with subsequent lamellar or penetrating keratoplasty being made with respect to a tumor type. The perfection of surgical technique consists in that one transplant of a bigger diameter is used instead of a corneoscleral transplant. PMID- 15529528 TI - [Prevention of complications in combined (laser and extrascleral) treatment of retinal detachment]. AB - The main factors of risk were analyzed and indications for preventive laser coagulation were specified in the paper. An approach was defined to ensure a maximally sparing surgical and stage-based laser treatment in retinal detachment, which is targeted at preventing its relapses and at improving the functional results. PMID- 15529529 TI - [Low-intensity helium-neon laser in the treatment of patients after endonasal dacryocystorhinostomy]. AB - Described in the paper is an efficiency study of using, postoperatively, the low intensity helium-neon laser (LIHNL) in patients with obstruction of the lacrimal tracts. Eighty patients were examined after endonasal dacryocystorhinostomy. They were shared between 2 groups with respect to a postoperative course: group 1- experimental, 40 patients, and group 2--control, 40 patients. The experimental patients received, apart from the traditional postoperative therapy, a course of LIHNL therapy. The controls received only the traditional postoperative treatment. The efficiency of postoperative treatment was evaluated by clinical, instrumental and laboratory examination methods. The study denoted that the use of LIHNL in the early postoperative period after endonasal dacryocystorhinostomy had a pronounced anti-inflammatory effect, speeded up the wound healing, prevented the growth of granulation tissues and the merging of the shaped lacrimal-sac fistula with the nasal cavity. LIHNL contributed to a complete recovery of the functional activity of the nasal mucous tunic. Thus, the LIHNL therapy essentially facilitates the postoperative management of patients, cuts the rehabilitation period and enhances the treatment results. PMID- 15529530 TI - [A comparative analysis of the inclination angle of rigid and soft IOLs with intracapsular fixation]. AB - Eighty-four eye were examined in 67 patients after phacoemulsification with intracapsular IOL implantation. The patients were shared, with respect to IOL rigidity, between 2 groups: group 1--rigid PMMA lenses and group 2--soft hydrogel and acryl lenses. The inclination angle was evaluated for different lens types by using an elaborated mathematical formula. The lenses had capsular fixation in all cases. The deviation angle ranged from 0.58 to 1.22 degrees. The difference between the inclination angle and ocular visual axis for rigid and soft lenses was statistically reliable (p < 0.05). The inclination angle of the optic axis in case of soft lenses versus rigid lenses was to a greater extent pronounced after 2 months of observation. However, the conclusion is not final due to continuous fibrosis of the capsular sac, which affects dynamically the IOL positioning. Therefore, similar studies will be made after 12-14 months to enable final conclusions. PMID- 15529531 TI - [The enzyme therapy within a complex treatment of hemophthalmos in patients with diabetes mellitus]. AB - The efficiency of the enzyme therapy was studied in patients with hemophthalmos concurrent with diabetes mellitus of different types. Sixty-eight patients received parabulbar injections of the hemase and collelizin enzymes and another 25 patients underwent the routine therapy comprising injections of dicynone, emoxipin and heparinum. The method of ultrasound B-scanning was used to determine the area of hemorrhage spread into the vitreous body. A high efficiency of the enzyme therapy, as used within the complex treatment of hemophthalmos in patients with diabetes mellitus, was demonstrated. Ultrasound B-scanning is a highly informative technique in evaluating the enzyme-therapy efficiency. The enzyme therapy can be recommended as the most effective method for the treatment of hemophthalmos in patients with diabetes mellitus. PMID- 15529532 TI - [Results of examination of the local immunity in demodectic blepharoconjunctivitis]. AB - Described in the paper are the results of examination of the local immunity in patients with demodectic blepharoconjunctivitis (DBC). Demodicosis of the eyes was established to develop in impairments of the local immunity described by a lower level of IL-4 in lacrimal fluid, a lower relative quantity of CD3+ cells, and a higher relative count of CD16+-lymphocytes infiltrating the eyelid conjunctiva epithelium. The age-related immunodeficiency manifested as a decreased relative number of CD3+ and CD4+-lymphocytes infiltrating the eyelid conjunctiva epithelium provokes a higher invasion of ticks and an increased morbidity of demodicosis among elderly people. DBC, when complicated by episcleritis or keratitis, is accompanied by a higher IgE level in lacrimal fluid and a decreased ratio of IgE/IL-4, which matters in prognostication and pathogenetic substantiation of DBC therapy. PMID- 15529533 TI - [The clinical-and-morphological specificity of using xenopericardium in plasty of the eyelids and orbit]. AB - The authors analyzed the clinical results of transplantation of the xenopericardium in 60 patients made at the Research institute of eye diseases of the Russian academy of medical sciences from 1999 to 2003. Xenomaterial was used in reconstruction of the lower eyelids after trauma (coloboma, defects of ligaments), in ectropions of the lower eyelids, in paralytic lagophthalmos and in traumas of the orbit. The postoperative follow-up of patients ranged from 2 to 54 months (an average of 28 months). Xenopericardia removed from patients (4 cases) were histologically examined in 6 months and more after surgery. The examination was made by the method of serial sagittal paraffin sections tinted with hematoxylin or eosin or according to I. van Gieson. A moderately pronounced tissue response and good remote clinical results confirmed the possibility of using it in reconstructive surgery, in particular, in plasty of the orbit especially when made through a small incision. The peculiarities of using clinically the xenopericardium denote that it can be used in ophthalmoplasty; however, it should be fixed immediately under the skin of the eyelids very carefully in order to avoid the formation of uneven surface. PMID- 15529534 TI - [Experimental intraocular introduction of blood serum of patients with endophthalmitis]. AB - Effects of blood serum from patients with endophthalmitis, who systematically received gentamicin at therapeutic dose, exerted on the retina of healthy rabbits were experimentally studied. A moderate toxic effect produced by gentamicin on the retina, when it was administered in the vitreous body of rabbits, was detected yet on day 1 of the experiment. Changes of retinal electrogenesis were aggravating and, beginning from day 7, there were noted profound changes of the functional activity in all retinal layers. According to the results, the blood serum from patients with endophthalmitis, who were systematically receiving gantamicin at therapeutic dose, does not virtually have any negative effect on the functional condition of the retina and can be used for intracavitary administration. PMID- 15529535 TI - [Experimental implantation of the "Ufalens" artificial lens in primary surgical treatment]. AB - Described in the paper is the response of eye tissues in extraction of traumatic cataract with IOL implantation observed in primary surgical treatment of penetrating corneal wounds. Experiments were made on 60 rabbits (120 eyes). 5 mm long penetrating corneal wounds located in the optic zone with damaged lens were modeled in the experimental group (30 rabbits, 60 eyes). The wound was surgically treated in 24 hours: traumatic cataract was extracted and posterior-chamber IOL was implanted ("Ufalens-1" manufactured at the UFA Research institute of eye diseases). Thirty rabbits (60 eyes) were in the control group: transparent lens was extracted in them and IOL (same model) was implanted. The animals were taken out of the experiment by air embolism on days 3 and 7 and in 1, 3 and 7 months. Tinting with hematoxylin or eosin was used to ensure plain histological examination. The stages of histological treatment of tissues were carried out according to the standard method. A conclusion was made on the basis of research results that the implantation of "Ufalens-1" in primary surgical treatment of eyeball wounds with damaged lens does not have any essential clinical or morphological impact on the severity of the postoperative period and can be recommended as a method of medical rehabilitation of victims for use at specialized ophthalmologic clinics. PMID- 15529536 TI - [A comparative evaluation of the oxidant and antioxidant blood systems in rabbits with caustic burns of the eyes in their treatment by synthetic and natural antioxidants]. AB - A method of provoking a caustic burn was described on the basis of experiments made in 36 rabbits (72 eyes) who were shared between 2 groups. Effects from histochrom, a natural antioxidant, and from emoxipin, a synthetic antioxidant, produced on the lipid-peroxidation parameters at early stages of ocular burn disease were studied. A higher clinical efficiency of histochrom versus emoxipin was demonstrated. PMID- 15529537 TI - [Measurement method and determination accuracy of the critical frequency of light flashing]. AB - The influence of a measurement method on an accuracy of determining the critical frequency of light flashing was investigated. The least incidental contributing error of measurements was found for the method based to be on progressive approximation with discrete changes of the frequency of light flashing made at the final measurement stage. PMID- 15529538 TI - [Life quality of patients with primary open-angle glaucoma]. PMID- 15529539 TI - [Dacryocystorhino-drainage as a treatment method in dacryocystitis]. PMID- 15529540 TI - [Gun-powder injury of the eye]. PMID- 15529541 TI - [Remote results of photorefractive keratectomy by MEL-60 excimer laser]. PMID- 15529542 TI - [A case of idiopathic uveal effusive syndrome]. PMID- 15529543 TI - [A comparative analysis of the efficiency of different methods in the treatment of amblyopia]. PMID- 15529544 TI - [Ocular signs of Maroteaux-Lamy syndrome]. PMID- 15529545 TI - [Influence of periodic light effects on the parameters of the ocular nystagmus]. PMID- 15529546 TI - [Prognostication of visual functions in modern surgery of cataract]. PMID- 15529547 TI - [Free-radical oxidation in the pathogenesis of eye diseases]. PMID- 15529548 TI - [Age-related insufficiency of accomodation (presbyopia): terminology, origin and principles of correction]. PMID- 15529549 TI - [Professor V. P. Odintsov--Director of the Eye Clinic of Moscow University and editor-in-chief of "Russiona Ophthalmologic Journal"]. PMID- 15529550 TI - Oxidized LDL and coronary heart disease. AB - During inflammation, several cell types synthesize and secrete phospholipase A2 that catalyses lipid oxidation in LDL. Myeloperoxidase, a haeme protein secreted by activated phagocytes, oxidizes L-tyrosine to a tyrosyl radical that is a physiological catalyst for the initiation of lipid oxidation in LDL. Lipid oxidation results in the generation of aldehydes that substitute lysine residues in the apolipoprotein B-100 moiety. Lipid together with protein oxidation in LDL results in the generation of oxidized LDL. We, among others, have demonstrated an association between coronary heart disease (CHD) and increased plasma levels of oxidized LDL. Recently, we have demonstrated a higher prevalence of elevated oxidized LDL in persons with high-calculated CHD risk prior to events. The odds of having elevated oxidized LDL for persons with high-calculated CHD risk prior to events were even higher than for persons with diagnosed CHD. A likely explanation is that once CHD has been diagnosed the patients are more treated with a statin that appears to decrease oxidized LDL even beyond its cholesterol lowering effect. We have identified several metabolic syndrome components (high triglycerides, low HDL-cholesterol, glucose intolerance and diabetes) that independently of LDL-cholesterol, predicted high levels of oxidized LDL. Finally, elevated oxidized LDL predicted myocardial infarction in the Health ABC cohort consisting of well-functioning elderly people, even after adjusting for age, gender, race, smoking, and the metabolic syndrome. PMID- 15529551 TI - Amlodipine and glutathione cycle in hypercholesterolaemia. AB - OBJECTIVE: Effects of amlodipine on lipid peroxidation and alterations in glutathione and related enzymes in blood and aortic tissue were investigated in a cholesterol-induced atherosclerotic rabbit model. METHODS AND RESULTS: New Zealand white male rabbits were fed with regular chow (group I), chow supplemented with I% cholesterol (group II), regular chow plus amlodipine 5 mg/kg/day p.o. (group III) and I% cholesterol diet supplemented with amlodipine (group IV) for 8 weeks. Cholesterol, malondialdehyde (MDA), reduced glutathione (GSH), oxidized glutathione (GSSG), glutathione peroxidase (GSH-PX) and glutathione reductase (GSH-Rd) were determined in blood samples drawn before and after the experimental period. Aortic tissue was examined morphologically for atherosclerotic changes and tissue cholesterol, MDA, GSSG, GSH-PX, GSH-Rd and glutathione-S-transferase (GST) were measured. After 8 weeks, blood cholesterol, MDA, GSSG and GSH-PX were elevated in groups II and IV; GSH was reduced in group IV; MDA levels were higher in group II than in group IV. Aortic tissue investigations revealed higher cholesterol and MDA concentrations in group II than in group IV. Morphological examination of aortic tissues exhibited endothelial disarrangement and lipid deposition in group II. Histopathological alterations related to atherogenesis were less in group IV than in group II. CONCLUSIONS: Amlodipine reduced the increase in oxidative stress by inhibiting excessive MDA production. Accelerated glutathione redox cycle activity of erythrocytes from animals supplemented with amlodipine suggests that this drug may reduce oxidative stress by enhancing the glutathione system. However, this drug does not seem to affect the glutathione redox cycle in the aortic tissue. PMID- 15529552 TI - Angiotensin-converting enzyme I/D gene polymorphisms and effects of left ventricular hypertrophy in Turkish myocardial infarction patients. AB - OBJECTIVE: Since the initial report of the association of the deletion/insertion (D/I) polymorphism in the gene for angiotensin-converting enzyme (ACE) with myocardial infarction (MI), there has been considerable controversy. Some have found the D allele to be associated with MI, coronary heart disease (CHD) or other cardiac pathologies, while others have not. In view of the clinical importance of the ACE as a major marker of cardiovascular diseases, we investigated the I/D polymorphism of the ACE gene in Turkish patients with acute myocardial infarction in comparison with control subjects. METHODS AND RESULTS: Polymerase chain reaction, and agarose gel electrophoresis techniques were used to determine the ACE genotype in 214 subjects. The frequencies of ACE D and ACE I allele among the patients with acute myocardial infarction were 65.54% and 36.45% and in the control subjects 57.62% and 42.37%, respectively. ACE DD genotypes were found higher in patients with left ventricular hypertrophy (LVH) than without LVH (55.6% vs. 37.7%; X2: 2.534, p > 0.05). CONCLUSIONS: The ACE D allele is more frequent in patients with acute myocardial infarction than in controls. Moreover ACE DD genotype might be associated with an increased risk of left ventricular hypertrophy. PMID- 15529554 TI - Changes of homocysteine levels and arterial endothelial function in patients with high risk of coronary artery disease after 6-month folic acid supplementation. AB - OBJECTIVES: This study aimed to assess whether folic acid supplementation could produce longer-term (6-month) improvements in homocysteine levels and arterial endothelial function in patients with a high risk (3 or more traditional risk factors) of coronary artery disease (CAD) and hyperhomocysteinaemia. METHODS: Thirty-one adults with 3 or more traditional risk factors of CAD and hyperhomocysteinaemia were selected, without CAD (the criterion of CAD is that more than one main vessel has an obstruction > or = 50%) by coronary angiography. All subjects were given folic acid (5 mg/day) for 6 months. Fasting plasma homocysteine levels were measured by high-performance liquid chromatography. Plasma folic acid and vitamin B12 levels were measured with immunoassay. Arterial endothelial function was measured as flow-mediated dilation of the brachial artery using high-resolution B-mode ultrasound. RESULTS: Folic acid supplementation for 6 months was associated with a significant increase in mean (+/- SD) plasma levels of folic acid (4.6 +/- 1.4 microg/l to 9.1 +/- 2.5 microg/l; P < 0.01) and a significant decline in homocysteine levels (18.3 +/- 3.9 micromol/l to 11.5 +/- 2.8 micromol/l; P < 0.01). Flow-mediated dilation also improved significantly, from 6.8% +/- 2.1% to 8.9% +/- 1.7% (P < 0.01). CONCLUSION: These results demonstrate that long-term (6-month) folic acid administration significantly declines homocysteine levels and improves arterial endothelial function and has potential implications for the prevention of atherosclerosis in adults with 3 or more traditional risk factors of CAD and hyperhomocysteinaemia. PMID- 15529553 TI - HLA-DR B1 and DQ B1 polymorphisms in patients with coronary artery ectasia. AB - OBJECTIVES: The purpose of our study was to evaluate the significance of polymorphisms in HLA class II genes in coronary artery ectasia (CAE) patients. METHODS AND RESULTS: Twenty-six patients with CAE without associated cardiac defects were enrolled in the study. CAE was defined as luminal dilation of 1.5- to 2.0-fold of normal limits. Ninety-five healthy subjects who were donors for different organ transplantations, were chosen as control group. Physical examination, electrocardiography and chest X-ray were completely normal in these cases. Both the patients and the control group were screened and compared for their HLA class II genotypes. HLA-DR B1*13, DR16, DQ2 and DQ5 genotypes were significantly more frequent in the patient group. When the known risk factors of coronary heart disease were compared in the patients carrying these genotypes with the non-carrying group, no significant differences were encountered. CONCLUSIONS: HLA-DR B1*13, DR16, DQ2 and DQ5 may be associated with the pathogenesis and increase the risk of CAE. PMID- 15529555 TI - Tumour necrosis factor-alpha in diastolic dysfunction. AB - BACKGROUND: Active relaxation develops as a result of sequestration of calcium into the sarcoplasmic reticulum, and is controlled mainly by sarcoplasmic reticulum calcium ATPase (SERCA) and phospholamban.Tumour necrosis factor-alpha (TNF-alpha) downregulates both of these proteins, so it may play a role in the development of abnormal relaxation. However, a possible relationship between TNF alpha and diastolic dysfunction has not been sufficiently evaluated in vivo. We investigated whether circulating levels of TNF-alpha increased in patients with relaxation abnormality. METHODS: Forty hypertensive patients with normal left ventricular systolic function were enrolled in the study. Age-adjusted values of echocardiographically measured mitral inflow velocities, E-wave deceleration time and isovolemic relaxation time were used to define normal and abnormal relaxation. Twenty of the patients (mean age 59.2 +/- 10.6) had a relaxation abnormality (group I), and the twenty other patients (mean age 45.9 +/- 7.9) had a normal diastolic function (group II). TNF-alpha levels were measured by ELISA. RESULTS: There were no significant differences between the two groups in terms of interventricular septal thickness, posterior wall thickness, left ventricular mass, ejection fraction, plasma creatinin level, and medication. Patients with a relaxation abnormality were older than those with a normal diastolic function (p < 0.001). TNF-alpha levels were similar in both groups (62.1 +/- 46.0 pg/ml for group I, and 48.7 +/- 51.4 pg/ml for group II, p = 0.089). CONCLUSION: In this preliminary study, we demonstrated that TNF-alpha levels did not increase in patients with a relaxation abnormality. However, we think that a possible relationship between TNF-alpha and diastolic dysfunction should be clarified by further studies involving a larger number of patients with a wider spectrum of diastolic dysfunction. PMID- 15529556 TI - How does neonatal thymectomy affect the immune system? AB - BACKGROUND: The aim of this study was to determine the effects of neonatal thymectomy on the immune system in later life. METHODS AND RESULTS: Immune system tests were performed in 26 children at 1 year of age. Thirteen of them had been operated for transposition of the great arteries and had thymectomy in the same operation in the neonatal period. Thirteen control subjects were normal. Immune system tests including white blood cell count, lymphocyte count, T and B cells subgroups (CD2, CD4, CD5, CD7, CD8, CD16, CD20, CD22, CD56), mitotic reaction to phytohaemagglutinin in lymphocyte culture. White blood cell count and lymphocyte count were performed. In the statistical analysis, Mann-Whitney U and Wilcoxon rank sum W tests were used for both groups. Statistical significance was taken at a value of P < 0.05. There was no significant difference in mean white blood cell count, mean blastic transformation reaction of lymphocytes to phytohaemagglutinin, and CD7, CD4/CD8, CD20, CD22, CD56 ratios between the two groups (P > 0.05). Significant differences in mean lymphocyte number, and CD2, CD4, CD5, CD8, CD16 ratios between the two groups were defined (P < 0.05). CONCLUSIONS: In our study, it was noticed that mainly T lymphocyte subgroups were effected by neonatal thymectomy. Although no infection requiring therapy was seen in the thymectomized patients, we advise to limit total thymectomy as much as possible in neonatal heart operations. PMID- 15529557 TI - Plasma interleukin-6 levels are increased in coronary artery ectasia. AB - OBJECTIVE: Recent studies have suggested a cytokine-induced tissue inflammation in the pathogenesis of abdominal aortic aneurysms and it has been documented that circulating interleukin-6 (IL-6) levels in these patients are increased. The aim of this study was to investigate whether a similar association also exists for patients with coronary ectasia, which may also be regarded as an abnormal dilatation of the arterial system. METHODS AND RESULTS: The study group was composed of 43 patients with coronary ectasia and 48 patients with normal coronary arteries constituted the control group. Coronary diameters were measured by quantitative angiography. A coronary diameter index was defined for each segment as the coronary diameter divided by the body surface area (BSA). A coronary segment with a diameter index of more than 1.5 fold of the control group was defined as ectatic. Baseline characteristics of the two groups were similar. Serum IL-6 levels were significantly higher in patients with coronary ectasia (5.18 +/- 2.04 pg/ml vs. 4.13 +/- 0.5 pg/ml, p = 0.002). There was no significant correlation with the maximal diameter of the most dilated coronary segment and IL 6 levels in patients with coronary ectasia (r = 0.10, p = 0.50). CONCLUSIONS: Results of this study have demonstrated increased levels of circulating IL-6 in patients with coronary ectasia which might indicate a possible role of inflammatory processes. Absence of a significant correlation between the dimensions of the ectatic segments and IL-6 levels might be due to the narrower range of the diameters of the coronary arteries compared with the abdominal aorta. PMID- 15529558 TI - Prospective randomized trial of transthoracic versus low-energy transvenous internal cardioversion in persistent atrial fibrillation. AB - OBJECTIVE: Electrical cardioversion (CV) is used to restore sinus rhythm (SR) in patients with atrial fibrillation (AF). In this prospective randomized study, we compared two different methods of electrical CV, namely transthoracic (TT) and low-energy transvenous internal CV (ICV), in patients with persistent AF with respect to efficacy, safety and the magnitude of myocardial damage provoked by either method. METHODS AND RESULTS: Fifty-two patients with persistent AF were randomly assigned to either TT (n = 26) or ICV (n = 26). The baseline characteristics of the two treatment groups were similar. TT CV was performed under sedation with hand-held electrodes in the apex-anterior position and high energy (100-360J) monophasic shocks. ICV was performed by a single catheter approach utilizing a special balloon-directed catheter with proximal and distal arrays of shock electrodes that were positioned in the right atrium and left pulmonary artery under fluoroscopy. Truncated, biphasic shocks of low energy (1 15 joules) were used. Cardiac troponin T (cTpnT), creatine kinase (CK) and CK-MB levels were assessed before and 24 hours after each procedure. SR was restored in 24/26 (92%) patients in the ICV and 22/26 (85%) patients in the TT CV groups (p > 0.05). The mean energy to achieve SR was significantly higher with the TT method (9.8 +/- 4.3 J vs. 246.4 +/- 73.6 J, p < 0.05). CV with either method caused no elevation in cTpnT levels. Total CK and CK-MB levels remained unchanged with ICV. On the other hand, TT CV resulted in a significant increase in total CK (51.8 +/- 30 vs. 156.5 +/- 255.3, p < 0.05) and a nonsignificant rise in CK-MB levels (14.7 +/- 7 vs. 17.3 +/- 1.1, p > 0.05). CONCLUSIONS: In this prospective randomized comparison, TT and ICV were found to be equally effective to restore SR in patients with persistent AF No evidence of myocardial damage was detected with either method. PMID- 15529559 TI - Haemoglobin level influences plasma brain natriuretic peptide concentration. AB - OBJECTIVE: It has been demonstrated that the haemoglobin (Hb) level is associated with the prognosis of congestive heart failure (CHF). Correction of anaemia has improved CHF outcomes even in patients without anaemia. Lower Hb level may play a more important role in left ventricular (LV) dysfunction than previously recognized. This study aimed to evaluate the association of Hb level with plasma brain natriuretic peptide (BNP) level as a marker of LV function adjusted for known determinants of BNP. METHODS AND RESULTS: Association of Hb level with plasma BNP level was studied in 279 outpatients of cardiology (mean age 61 +/- 16, 54% men) using multivariate regression analysis. Mean Hb level was 13.7 +/- 1.5 g/dl and 14% of patients had anaemia. Median BNP level was 28 pg/ml (range < 4 to 580 pg/ml). In total subjects, the multivariate model adjusted for age, sex, history of CHF, atrial fibrillation, serum creatinine level, LV wall motion abnormality, end-diastolic LV dimension, LV mass index, and cardiovascular risk factors showed that a lower Hb level was significantly associated with higher BNP level (p = 0.0243). In "normal" subjects who did not have a history of CHF, atrial fibrillation, LV wall motion abnormality, LV dilatation, valvular abnormality, or LV hypertrophy, a lower Hb level was significantly associated with a higher BNP level (p = 0.0012) after adjustment for age, sex, serum creatinine level, and cardiovascular risk factors. CONCLUSIONS: Lower Hb levels are associated with higher plasma BNP levels independent of age, sex, serum creatinine level, LV wall motion abnormality, LV hypertrophy, history of CHF, atrial fibrillation, and cardiovascular risk factors. PMID- 15529560 TI - Circulating white blood cell count correlates with left ventricular indices independently of the extent of risk area for myocardial infarction after successful reperfusion. AB - OBJECTIVE: To test the hypothesis that the circulating white blood cell (WBC) and neutrophil counts are related to left ventricular (LV) indices in patients with the same risk area for acute myocardial infarction (AMI), we examined 100 consecutive AMI patients who had the culprit lesion at segment 6 according to the American Heart Association classification and who underwent successful direct coronary angioplasty. METHODS AND RESULTS: The LV ejection fraction (LVEF), end systolic volume (LVESVI) and end-diastolic volume index (LVEDVI) were obtained by left ventriculography performed 4 weeks after AMI onset. Univariate analysis disclosed that the counts of WBC and neutrophils on admission, and the maximal WBC count correlated negatively with LVEF (r = -0.46, p < 0.001; r = -0.54, p < 0.001 and r = -0.40, p < 0.001, respectively) and positively with LVESVI (r = 0.43, p < 0.001; r = 0.55, p < 0.001, and r = 0.30, p < 0.01, respectively). The counts of WBC and neutrophils on admission also correlated with LVEDVI (r = 0.28, p < 0.01 and r = 0.41, p < 0.001, respectively). Multivariate analysis with other clinical and angiographic factors revealed that the counts of WBC and neutrophils on admission correlated with LVEF (partial correlation coefficient, r = -0.37, p < 0.001 and r = -0.52, p < 0.001, respectively), with LVESVI (r = 0.34, p < 0.01 and r = 0.56, p < 0.001, respectively) and with LVEDVI (r = 0.28, p < 0.01 and r = 0.44, p < 0.001, respectively). The maximal WBC count also correlated with LVEF and LVESVI (r = -0.40, p < 0.001 and r = 0.21, p < 0.05, respectively). CONCLUSION: The present study revealed that the circulating WBC count correlated with function and volume of the successfully reperfused LV after AMI in patients with the same risk area for AMI, indicating that the WBC count needs to be taken into consideration as an independent factor affecting the LV indices. PMID- 15529561 TI - 99mTc-MIBI SPECT assessment of the effects of aneurysm resection on the left ventricular morphology. AB - OBJECTIVE: 99mTc-MIBI SPECT is a widely used myocardial perfusion investigation technique, but few data are available concerning its use to assess the morphological characteristics of a left ventricular aneurysm (LVA) before and after LVA resection. METHODS AND RESULTS: Pre- and postoperative rest 99mTc-MIBI SPECT images were analysed in order to characterize the features of LVAs and the changes in the 3D scintigraphic parameters after apical LVA resection in 6 patients. In the middle horizontal slice an angle was defined to quantify the apical divergence associated with the LVA. After resection, the changes in the divergence angles (DA) were measured as were the changes in the left ventricular volumes (LVV) by volumetric calculations. The mean DA decreased from an average of 38.50 degrees +/- 11.32 degrees preoperatively to 24 degrees +/- 11.84 degrees postoperatively (p = 0.03). The mean LVV also decreased significantly: from 443 +/- 87 ml to 317 +/- 74 ml (p = 0.003). The resectable LVAs were associated with a very low isotope uptake in the apical segments (< 20% relative activity). A DA < 20 degrees was also characteristic of anatomical LVA in all patients. A regression curve plotting divergence angle and the number of left ventricular segments below 20% relative activity showed a significant correlation between them (r = 0.86, p = 0.003). CONCLUSIONS: The significant decreases of DA and LVV after resection reflect favourable morphological changes in the left ventricle (reverse remodelling). We consider 99mTc-MIBI SPECT a useful method for apical LVA detection, it allows an analysis of the morphological (and indirectly the functional) results of the surgery. PMID- 15529562 TI - Role of statins in acute coronary syndromes. AB - Statins reduce major coronary events and all-cause mortality in patients with coronary artery disease. Statin therapy has a proven track record for the secondary prevention of coronary artery disease. The extension of the benefit to patients with acute coronary syndromes can be expected. Apart from their lipid lowering effects, statins significantly and favourably alter the natural history of acute coronary syndromes. Promotion of plaque stability, improvement of endothelial dysfunction and reversal of coagulation and platelet abnormalities are only some of the effects which are beneficial in the management of these patients. Early potential clinical benefits further strengthen the rationale for starting statin treatment as soon as possible after acute coronary events particularly in patients in whom invasive intervention is planned. This review examines their role in acute coronary syndromes and emphasizes the benefits of early statin therapy. PMID- 15529564 TI - Permanent ventricular pacing via a low posterolateral cardiac vein in a patient with a mechanical tricuspid valve prosthesis and complete atrioventricular block. AB - In the presence of an artificial tricuspid valve, permanent pacing is generally achieved with an epimyocardial approach. This report describes the implantation of a dedicated pacing electrode in a low posterolateral cardiac vein in a patient with a mechanical prosthetic tricuspid, mitral and aortic valve in whom an epimyocardial lead and pacemaker generator had to be removed because of infection. PMID- 15529563 TI - Different effects of antihypertensive drugs on endothelial dysfunction. AB - Since endothelial dysfunction may significantly contribute to the pathophysiology of hypertension and its complications, its modification seems to be a very attractive means to favourably affect the development of atherosclerosis and cardiovascular events in hypertensive patients. However, not all antihypertensive drugs consistently improve endothelial dysfunction. While first-generation beta blockers showed contrasting or null effects on endothelial function, newer beta blockers of the third generation, such as carvedilol and nebivolol, seem to be provided with specific endothelium-mediated vasodilating effects. Calcium channel blockers are generally able to increase endothelium-dependent vasodilation in several vascular beds, in patients with essential hypertension, probably through multiple mechanisms. Most studies have shown thatACE inhibitors favourably affect endothelial function mainly in the subcutaneous, epicardial and renal circulation, not only by inhibiting the effects of angiotensin II on the endothelium, but also by enhancing bradykinin-induced vasodilation, probably a hyperpolarization-related effect. On the other hand, discordant evidence is available about the effects of angiotensin II receptor type I blockers on endothelial function in patients with essential hypertension, atherosclerosis or diabetes.There are data suggesting that an increased activity of the endothelin- I system may play a role in the blunted endothelium-dependent vasorelaxation of hypertensive patients, an effect that could be contrasted by the use of endothelin-I receptor antagonists. However, to date no substantial clinical efficacy of endothelin-I receptor blockers has been shown in patients with essential hypertension. Finally, other possibly useful compounds in restoring impaired endothelial function in hypertension are some antioxidant agents such as vitamin C, folic acid, the cofactor tetrahydrobiopterin (BH4), L-arginine and the drugs of the statin class. PMID- 15529565 TI - Migraine headache induced recurrent atrial fibrillation: a case report. AB - A variety of symptoms that accompany migraine are mediated by the autonomic nervous system. The autonomic nervous system has long been known to play a potentially important role in the occurrence of atrial fibrillation. Clinically significant effects on cardiac rhythm are uncommon during migraine attacks. In the present case, we report a man who experienced recurrent episodes of atrial fibrillation, which were likely secondary to the vomiting component of migraine. PMID- 15529566 TI - Relationship between acute pancreatitis and ACE inhibitors. AB - Most of the acute pancreatitis cases are due to cholelithiasis and alcohol intake. Two percent of the cases are related to medications. Drugs including ACE (angiotensin-converting enzyme) inhibitors and thiazide diuretics may cause acute pancreatitis. Patients with biliary system disorders using certain drugs may develop acute pancreatitis and this may be confusing when considering the aetiology. We report a patient without any known risk factor who developed acute pancreatitis shortly after the intake of lisinopril and hydrochlorothiazide. Common bile duct stone and biliary sludge were diagnosed after physical and radiological evaluation. PMID- 15529567 TI - Coronary artery fistula in a diabetic patient with multiple vascular and other complications: a case report with review. PMID- 15529568 TI - Syncope due to autonomic insufficiency syndromes associated with orthostatic intolerance. AB - Recurrent syncope may be either a sign or a symptom and may occur due to a wide variety of very different causes. Extensive investigations into the nature of this disorder soon uncovered that it represents only one aspect of a broad, heterogenous group of disturbances of the autonomic nervous system (ANS) that can result in hypotension, orthostatic intolerance, and often syncope. Disorders of orthostatic regulation may be subgrouped into both primary and secondary forms. In primary autonomic failure syndromes, as opposed to the intermittent periods of hypotension seen in the reflex syncopes, patients could develop orthostatic intolerance due to a failure of the ANS to function under normal circumstances. Chronic autonomic insufficiency has two entities: Pure Autonomic Failure (PAF) and Multiple System Atrophy (MSA). Over the last several years, it has become apparent that a milder form of autonomic insufficiency occurs that is now referred to as the Postural Orthostatic Tachycardia Syndrome (POTS). The secondary forms of autonomic failure occur in association with a particular disease process. One of the most important things to remember are the vast number of pharmacologic agents that may either cause or worsen orthostatic hypotension. The principal feature that all of these conditions share is that normal cardiovascular regulation is disturbed resulting in postural hypotension. The comerstone of evaluation is a detailed history and physical examination. One of the physician's most important tasks is to identify whether hypotensive syncope is primary or secondary in nature, and to determine if there are any potentially reversible causes (i.e., drugs, anemia, volume depletion). It is equally important to educate the patient. Nonpharmacologic therapies are useful. Pharmacotherapy should be used cautiously in selected cases. PMID- 15529569 TI - The treatment of chronic hepatitis B. AB - The diagnosis of chronic hepatitis does not necessarily imply a treatment. This statement is endorsed by the fact that many forms of the disease are non-evolving or very slow evolving, and therefore development of cirrhosis or hepatocarcinoma is extremely unlikely in these minimal and mild forms of the disease. Furthermore, drugs used today occasionally lead to virus annihilation and, in any case, do not operate upon virus integration into the hepatocyte genome. However, at present there is a consensus regarding the opportunity of therapeutical intervention in chronic hepatitis B with major biological and histological signs of activity, although there remain two important issues to approach. The first one is the relatively low efficacy of therapy referred to the cost (i.e. a maximum of 40% of cases with long-lasting therapeutic response to interferon), while the second is the presence of various side effects, which can force the reducing of dosage or even the discontinuation of treatment. Antiviral therapies, which were using acyclovir, didanosine, zidovudine and vidarabine, have demonstrated their inefficiency. Therefore, two groups are still used nowadays: interferon and nucleosides analogues. Each of the medication enounced has defined dosages, treatment extent, indications and contraindications, side effects and therapy expectancies. Proper management in chronic hepatitis B can achieve the following end-points: obtaining HBeAg-to-anti-HBe seroconversion, prevention of the disease progression and quality-of-life improvement. The treatment of chronic hepatitis B is an ever-changing issue and our expectation for better results grows proportionally with the knowledge brought everyday by new research and clinical experience. PMID- 15529570 TI - Role of Nad(P)h: quinone oxidoreductase in the regulation of intracellular redox state. AB - This paper is a brief overview of current knowledge about DT-diaphorase [NAD(P)H: Quinone Oxidoreductase, NQO], flavoprotein that catalyzes the obligatory two electron reduction of a wide variety of substrates. The most efficient substrates are quinones but the enzyme will also reduce quinone-imines, nitro and azo compounds. NQO is unique among known NAD(P)H-oxidizing flavoproteins in being a 2 electron transferring quinone reductase, and play a major role in preventing one electron reduction of exogenous quinones by other enzymes to auto-oxidable semiquinones and concomitant superoxide-radical generation. Induction of NQO by a variety of xenobiotics (potential sources of free-radical formation which lead to DNA and cell damage) provides protection from the cytotoxic and carcinogenic effects of these compounds. NQO has an important role in the bioreductive activation of various quinones used in cancer chemotherapy. PMID- 15529571 TI - Chlamydia pneumoniae antibodies and inflammatory reaction in patients with ischemic heart disease. AB - We studied the relationship between Chlamydia pneumoniae antibodies, C-reactive protein (CRP) and interleukine 8 (IL-8) in 87 patients with ischemic heart disease: 29 patients with acute myocardial infarction, 18 patients with unstable angina pectoris and 40 patients with stable effort angina. We determined in all patients IgG and IgA antibodies to Chlamydia pneumoniae, CRP and IL-8. Species specific antibodies to Chlamydia pneumoniae (IgG and IgA) were detected by indirect ELISA technique (Savyon Diagnostics Ltd, Israel). Interleukine-8 measured by a commercially available ELISA kit (CLB, Amsterdam, The Netherlands). CRP was determined by radial immunodiffusion (Mancini). The IgG antibodies were present in 25 patients (29%), the greatest percentage being noted in patients with unstable angina pectoris (50%). The IgA antibodies were present, as a sign of chronic Chlamydia pneumoniae infection, in 56% of the patients with IgG antibodies. CRP was positive in 52% of the 25 patients with positive IgG antibodies, but in only 34% of the 62 patients without IgG antibodies (p < 0.01). IL-8 was positive in 12% of the patients with IgG antibodies, and in 21% of the patients without IgG antibodies but the difference is not significant (p > 0.05). It is concluded that there is a relationship between the presence of the Chlamydia pneumoniae infection and inflammatory reaction in patients with ischemic heart disease, but the neutrophils are not implied in this process. PMID- 15529572 TI - Study of substitution hormonal therapy on cardiovascular diseases in post menopausal women. A multicentric international collaboration (part I). AB - This is a collaboration begun in a multicentric international study, which proposed to follow (SHT) effects on cardiovascular diseases inpost-menopausal the substitution hormonal therapy women (PMW). OBJECTIVES: Feasibility study of SHT on Romanian PMW (from Bucharest) was based on English working protocol of the coordinator and on the good practice rules. We proposed to include in trial 100 150 PMW. We have solicited 456 women for study participation, age between 45-64 years, which have lesser than 1 year after the stop of menstruation and accepted all conditions to include in study (absence of mammary, uterine, or other cancers, deep venous phlebitis). All conditions were accepted by 121 women. Mean age was 55 +/- 5 years. METHODS: All PMW was randomized in many groups of study. It was administrated either active SHT (estrogens as Progynova, or estrogens and cyproteronate as Climen, both from Schering) or Placebo (Pl) in women grouped: 1. nonhysterectomized (93) randomized for Climen (42) or Pl (42), and hysterectomized (28) randomized for Climen (9), Progynova (9), Pl (10). The effectuated investigations were: clinical history; laboratory data for blood lipids, coagulation, inflammation, cytovaginal control; mammography; osteodensitometry DEXA. Cardiology investigations were: ECG, exercise ECG, echocardiography and Holter registrations in selected case. Control examinations were made at 3 months mean period, excepting mammography and DEXA. RESULTS: After 3 months 46 PMW (38%) retired from the study for: altered first mammography 18 (15%), for side drug reactions (only for Progynova) 8 (7%), for other reasons 20 (17%). CONCLUSIONS: This feasibility study shows that, in Romania, a clinical trial on cardiovascular and general effects of SHT in PMW is possible. But, for realization of the proposed aim of the study to follow up a minimum 600 women in 10 years, we estimate the need of recruitment of 2500-3000 women, which must conform to the protocol conditions. PMID- 15529573 TI - Feasibility study of substitution hormonal therapy on cardiovascular diseases in post-menopausal women (part II). AB - BACKGROUND: In a finished feasibility study concerning the effects of substitution hormonal therapy (SHT) on the cardiovascular status in the post menopausal women (pmw) we analysed the rate of participation of the Romanian women, the reasons for abandon to participation, etc. in a multicentric international study. OBJECTIVES: This study had intention to analyse the cardiovascular status of the pmw proposed for STH. MATERIAL AND METHODS: Only 121 women, who accepted all conditions for participation to study, were included. Mean age was 55 +/- 5 years (45-64 years), 93 women were in natural, and 28 in surgical menopause. The study protocol included analysis of: 1) the coronary risk factors (arterial hypertension, diabetes, smoking, serum lipids level, etc.), 2) inflammatory syndrome (serum fibrinogen, C reactive protein), 3) cardiovascular status (clinical data, ECG). RESULTS: 1) Incidence of essential hypertension was 23%; type II diabetes 17%; smoking 15%; serum high cholesterol was presented in 82%; high lipids level in 56%; high serum triglycerides in 35%; HDL was reduced in 56%. 2) Serum high fibrinogen was detected in 20% and CPR presence in 20%; 3) ECG was altered in 30% (HVS, ST-T modifications). Only 20% from pmw with alterations of ECG have not presented coronary risk factors or inflammatory syndrome. DISCUSSION: It is a feasibility study in a domain very extended actually by many multicentric clinical trials. The problem of the effects of SHT on cardiovascular system is still in dispute, and our study estimates the preliminary data at the beginning of a trial. These data show a high percent of pmw, which have already alterations of ECG and a concurrency of coronary risk factors and inflammatory indices. The effects of SHT in these women necessitate still a study on long time interval. For Romanian pmw, these data indicate the importance of the precocious investigations and detection of all coronary risk factors concerning the correct indications of SHT and its effects. PMID- 15529574 TI - Lectin-binding profile of serum IgA in women suffering from systemic autoimmune rheumatic disorders. AB - In many pathological states changes in the carbohydrate part of serum glycoproteins occur, the etiopathogenic role or diagnosis significance of these abnormally glycosylated glycoproteins being relevant in many instances. The aim of this study was to determine the glycosylation pattern of the serum immunoglobulin A (IgA) in young women suffering from systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), by comparison with that in healthy ones. We used lectin proteins as bivalent reagents to selectively and specifically recognize and bind the different oligosaccharides at the surface of the IgA molecules and precipitate them. The serum IgA level in the SLE and RA women included in this study was of 4 and respectively 6 times greater than in control women. Our experiments revealed for the IgA isolated from SLE women a very low degree of glycosylation, as follows: in the Fc region, we detected a markedly reduced level of the unbisected, biantennary oligosaccharides (by 72.8%) and of the unbisected, tri- and tetraantennary oligosaccharides (by 40%), accompanied by a slightly increased level of the bisected (by 14.7%) and fucosylated (by 13.3%) oligosaccharides; in the hinge region, a marked degalactosylation (by 74.4%) of the oligosaccharides was revealed; the sialylation degree of the IgA molecules was thus reduced by 71.2%. Related to it, it is reasonable to presume a certain role of the degalactosylated IgA fraction in renal tissue deposition of IgA in SLE patients, as it has been ascribed to this fraction in IgA nephropathy. Referring to the IgA isolated from RA women, we showed that it selectively bound the lectins, but lacked the property to be lectin-precipitated, most probably due to an altered arrangement of the oligosaccharide chains at the surface of the glycoprotein. PMID- 15529575 TI - Serological levels of apoptotic bodies, sFAS and TNF in lupus erythematosus. AB - In our study we have investigated the presence of apoptotic bodies, soluble FAS receptor and TNF (tumor necrosis factor) in three clinical forms of lupus erythematosus. Determinations were performed in attack period of: systemic lupus erythematosus (SLE) for 20 patients, 20 patients with subacute cutaneous lupus erythematosus (SCLE), 20 patients with chronic discoid lupus erythematosus (DLE). Determinations were performed by ELISA (for apoptotic bodies, kit Boehringer, normal values 400-800 mU), (for sFAS, kit R&D Systems, normal values 4500-17000 pg/ml) (for TNF, ELISA kit R&D Systems, normal values 0.4-3.6 pg/ml). Results in SLE: apoptotic bodies were increased in 16 cases (980-1030); sFAS in 18 cases (17000-24000 pg/ml) TNF was increased in all 20 cases (40-140 pg/ml). In SCLE with multiple cutaneous lesions and without internal organs disturbance the apoptotic bodies were increased in 10 cases (960-1030 pg/ml), sFAS in 9 cases (17000-22000 pg/ml), and TNF alpha in 9 cases. In DLE, apoptotic bodies were increased in 2 patients (980-1010 pg/ml), sFAS in 3 patients (17000-20000 pg/ml) and TNF in 2 patients (20-40 pg/mil). Investigated values were slightly correlated with immune parameters (anti dsDNA antibodies), but they were correlated with the presence of renal disturbances or extension of cutaneous lesions. We consider that the presence of increased apoptotic bodies as a result of peripheral mononuclear cells apoptosis appear as a nauto-limiting mechanism in a pathological immune response. The increase of sFAS in lupus patients serum might be interpreted as an alteration of apoptosis respectively a deficit in apoptosis which has as a first consequence the persistence of B and T lymphocytes, activated, in the pathogen immune response. PMID- 15529576 TI - Hand abnormalities of the patients with diabetes mellitus. AB - Hand abnormalities, represented by Dupuytren's contracture, limited joint mobility, carpal tunnel syndrome, flexor tenosynovitis,occur in approximately 50% of the diabetic patients, affecting their activity and decreasing the quality of their life. Using specific methods, these can by detected long before they are observed by the patients. These disturbances have a high prevalence inthe persons with long diabetes mellitus duration, the prevalence increasing with the age. Smoking, usual hard activities, bad metabolic control and so on, can increase the severity of these disturbances. It is compulsory a careful examination of the patients' hands, because more abnormalities can be surgically rightened. PMID- 15529577 TI - Serum level of soluble intercellular adhesion molecule-1 in patients with acute viral hepatitis A or acute viral hepatitis B: lack of correlation with severity of the disease. AB - Serum soluble ICAM-1 (sICAM-1) was found to be increased in patients with chronic hepatitis. The aim of the study was the evaluation of sICAM-1 dynamics in patients with acute hepatitis A or hepatitis B. Serum level of sICAM-1 was determined in 13 patients with acute hepatic A, 19 patients with acute hepatitis B, and 20 healthy controls. Measurements were done four times, at the beginning of jaundice, in the 14th day of jaundice, at release from the hospital, and when biochemical indices of the liver function became normal. An increase in sICAM-1 was shown in the patients. There was no difference between patients with hepatitis A and those with hepatitis B. The sICAM-1 decreased in the course of recovery but the values in the last measurements were still higher than in the controls. There was no correlation of sICAM-1 and duration of hospitalization. The obtained results do not suggest sICAM-1 as diagnostic or prognostic marker in patients with acute hepatitis A or B. PMID- 15529578 TI - Nuclear magnetic resonance study of changes induced by the energy of ultrasonic field in the lungs. AB - 156 laboratory mice were exposed to ultrasonic (US) waves under 12 different conditions. After exposure to ultrasounds, the left lung of each animal was studied morphopathologically and then analysed by nuclear magnetic resonance (NMR) spectroscopy. The study emphasized the possibility of hemorrhagical lesions in the lung through US and the fact that both measurement techniques (for T1 and T2) are alike in sensitivity. The effect on the lung is depending on the intensity and time of exposure to US. PMID- 15529579 TI - Study of anti-striational and anti-thyroid antibodies in patients with myasthenia gravis. AB - The study was carried out in 74 patients out of whom 46 with generalized myasthenia gravis and 28 with the ocular form of disease--all in acute state, ranging in Osserman's severity scale between I-IV; the age at onset of disease varied between 18-79 years. It showed anti-striational antibodies in 31% of the patients and anti-thyroid antibodies in 40%, as compared with 41 controls. Likewise it was observed that the presence of anti-striational antibodies with significant high values was associated with the presence of thymomas in 16 generalized myasthenia gravis patients and in 3 ocular myasthenia gravis patients (p < 0.001). Anti-thyroid antibodies were more frequently observed in ocular myasthenia gravis (23 cases) as compared to generalized myasthenia gravis (7 cases) with significant high levels (p < 0.001). The results obtained showed that the study of anti-striational antibodies can be of great utility in the diagnosis of thymoma, and that the study of anti-thyroid antibodies can be of great utility to sustain the clinical diagnosis of ocular myasthenia gravis. The presence of anti-striational and of anti-thyroid antibodies showed a basic alteration of immunoregulatory mechanisms of maintaining immune tolerance in myasthenia gravis patients. PMID- 15529580 TI - Leukotrienes antagonists in the management of bronchial asthma. AB - Leukotrienes antagonists represent the first new class of anti-asthma drugs available in the last two decennia, based on the great progress from medical research to clinical practice. They have been already intensively studied in many clinical trials and continue to be an important subject of debate. This article intends to summarize the most recent data published in the literature about the role of leukotrienes in the pathogenesis of asthma and the actions of leukotrienes antagonists, pointing out their benefits in the light of the concept of efficacy and effectiveness. PMID- 15529581 TI - Farmer's lung. AB - Farmer's lung is a well known occupational disease, described many years ago; the incidence in our country is still very low. That is the reason why we find interesting to make a review of the main diagnostic procedures and to present the new data in this form of hypersensitivity pneumonitis. Other farmers' occupational induced respiratory diseases are also presented, with their similitudes and their differences from the classical farmer's lung. PMID- 15529582 TI - The endothelial dysfunction in diabetes mellitus. AB - The vascular chronic complications are the main cause of morbidity and mortality in patients with diabetes mellitus. Nowadays it is well known the fact that the arteriosclerosis is initiated by the injury of the vascular endothelium and that the normal endothelial cells are producing a number of vasoactive factors. Thus, the vascular endothelium was considered an inert "lining" layer, but today is seen as a complex organ, with paracrin and autocrin function, which provides a "first line" physiological defence against atherosclerosis. Impaired endothelial function occurs in people with diabetes as a result of associated conditions (e.g. hyperglycemia, hypertension, dyslipidemia), or as an effect of hyperglycemia itself (e.g. cytokines, free fatty acids, AGES). The presence of endothelial dysfunction in non diabetic insulin resistant subjects suggests that metabolic and vascular abnormalities are tightly related at a fundamental level. Recent evidence suggests that insulin signalling for glucose transport in classical target tissues (muscle and adipose tissue) and upregulation of NO production in the endothelium utilises the same postreceptor pathway. This pathway involves the enzyme P13 kinase. Knowledge of the molecular mechanisms involved in the pathogenesis of endothelial dysfunction may ultimately result in novel approaches to the treatment and prevention of cardiovascular disease in people with diabetes mellitus. PMID- 15529583 TI - The use of light reflection rheography in diagnosing venous disease and arterial microcirculation. AB - The use of light reflection rheography (LRR) has reached an important level in the past few years. Using the originally construed and conceived RRL 001 device, we present the basic principles, the working technique and the limits of this method. We will use two magnetic amplifiers, one for the finger tip, which measures the capillary blood flow at this level and a venous magnetic amplifier which measures the blood flow in the venous plexuses at a 1-2 mm depth. The detection of the blood flow in the subcutaneous (hypodermic) venous areas leads to the diagnosis of profound thrombophlebitis and venous valves insufficiency. The detection of the capillary flow in the finger tip, its presentation as a curve, measuring the pressure of the inter digital arteries (using Riva Rocci method), establishing the index of systolic pressure make possible the understanding of the gravity of the existing hemodynamic disorders (especially in what concerns the Raynaud syndromes), the supervision of the original disease and the efficiency of the treatment. Weindorf was the first one to use the RRL technique in medicine in 1985 and, after 1997, it has been developed through Sproule's work. Within The Medical Clinic of the Municipal Hospital, a light reflection rheograph has been built and forwarded as innovation. The device has been used for 2 years in the diagnosis of venous disorders and peripheral microcirculation disorders. In this article we present the principles of the LRR examination of the lower limb venous system and of the microcirculation of the fingers of the upper limbs, an examination based upon our own experience which includes 72 examinations. PMID- 15529584 TI - Clinical particularities in a Romanian series of Behcet's disease patients. AB - Behcet's disease (BD) is characterized by a great geographic diversity of clinico evolutive features. We identify the main clinical characteristics in our series as compared to other data in international literature. We studied 36 patients (16 women and 20 men) with BD fulfilling International Study Group for Behcet's Disease (ISGBD) criteria. We recorded the clinical features and compared the data with other recent statistics. The incidence of clinical manifestations in our group was the following: oral ulcerations -97.4%, genital ulcerations - 55.5%, vascular disease - 50%, antiphospholipid antibodies - 55%, digestive tract lesions - 28.5%, eye disease - 27.7%, pulmonary disease - 8.3%, arthritis - 5.4%, CNS lesions - 2.7%. The clinical course was generally mild. Our patients had a higher incidence of vasculo-thrombotic events, of APLAs and of oral aphthosis, as compared to other statistics. Eye disease, CNS, articular involvement and pathergy were encountered rarer than in other groups. Two cases had atypical onsets: one with a pyoderma gangrenosum-like lesion and the other with severe pulmonary hypertension. BD has protean clinical aspects, with important geographic particularities. Our patients ran a relatively mild course of the disease, more like the Western European than the Asian patients. Vasculothrombotic disease was an important feature, with a subsequent risk for mortality. PMID- 15529585 TI - Clinical considerations concerning progressive diabetic renal diseases. Severe complications of early onset diabetes mellitus. AB - The main question of diabetic nephropathy in early onset diabetes mellitus consists of the possibility to advance to chronic renal failure at an early age, a complication having severe consequences in medical, familial and socio-economic life. The authors of this study are purposing to discover risk factors involved in the evolution of diabetic nephropathy (DN) with the aim to act by tertiary and secondary prophylactic measures which will decrease or stop the evolution of final forms of this disease. Our study contains 129 patients with insulino dependent diabetes mellitus (type 1), which started under twenty years age, screened between 1965-2000. The average age is 21 years and the average age of disease's onset is 12 years. We used Mogensen's classification in order to border the DN. The study refers only to the patent DN, called "OVERT-DN", which represents the fourth and the fifth stages. High arterial blood pressure values were taken into consideration when these are higher than 130/85 Hg mm. The informations were statistically analysed. DN affects 21% from the lot of 129 patients. We noticed differences between males and females from 80 type 1 DM males, 23.7% had DN and from 49 type 1 DM females 16.3% had DN. High arterial blood pressure appeared to 19.3% from all patients and as a secondary effect of DN in 18.6% of the cases; "border-line" essential high arterial blood pressure was found in only one patient. From the 27 patients with clinic DN, 44.4% evoluated to chronic renal failure (CRF) and half of them suffered till final stage of CRF. The frequency of DN was linked with the duration of diabetes. Thus, DN affects 13.8% of patients with 6-10 years of illness; 54.5 of patients with 20 years of illness, and after 25 years of illness - 87.5%. After 23 years of diabetes, it was accounted mortality in 3 of 7 cases, caused by final CRF. PMID- 15529586 TI - Evaluation of the metabolic balance in type 2 diabetes by assay of the hair glucose. AB - The authors used a relative new method of long term retrospective evaluation of metabolic balance in type 2 diabetes. This method was based on the variations of the glucose in the hair with the variations of HbA1c (r=+0.96; p<0.001). In this way the authors could follow up the diabetic's metabolic balance on longer time periods than by determination of the HbA1 values. PMID- 15529587 TI - Remission induction in elderly acute myeloid leukemia by low-dose melphalan. PMID- 15529588 TI - Effect of fasting (80h) on the luminol-enhanced chemiluminescence of the polymorphonuclear leukocytes in healthy human subjects. AB - It is already shown that fasting has antiinflammatory, antitumor and antiageing effects, but the mechanisms responsible for these are still unclear. An increased oxidative stress is one of the common traits for inflammation, carcinogenesis and ageing, so it is a reasonable assumption to consider that fasting might act through influencing the oxidative status. In the present study we used three different respiratory burst activators (opsonized zymosan-OZ, phytohemagglutinin P-PHA, concanavalin A-Con A) in order to test whether fasting is able or not to change the free radical generation and associated chemiluminescence produced by polymorphonuclear leukocytes (PMNLs) of seven healthy human subjects. Fasting has induced a "spontaneous respiratory burst" of resting polymorphonuclear leukocytes, increasing the basal chemiluminescence of these cells and it has decreased the stimulatory index of leukocytes. In conclusion, fasting has a dual effect on the free radical generation of polymorphonuclear leukocytes. PMID- 15529589 TI - Investigation of TNF-alpha, IL-6, IL-8 and of procalcitonin in patients with neurologic complications in sepsis. AB - Some mediators of inflammation are associated with sepsis, involving nervous system. Proinflammatory cytokines, TNF-alpha, IL-6, and IL-8, and procalcitonin (PCT), proinflammatory protein, were investigated in patients with neurologic complications in sepsis. TNF-alpha, IL-6, IL-8, and PCT were prospectively investigated in 62 patients with neurologic complications in sepsis. TNF-alpha and IL-6 were studied both in serum as in the CSF, IL-8 and PCT were studied only in serum. TNF-alpha, IL-6, and IL-8 were studied by ELISA (R & D Systems), and the PCT by immunoluminometric assay (BRAHMS). Mean value of TNF-alpha in serum was 578+/-214 pg/ml, and in CSF was 458+/-167 pg/ml (p<0.01). Mean value of IL-6 in serum was 749+/-213 pg/ml, and in CSF was 617.5+/-182 pg/ml (p<0.01). Mean value of IL-8 in serum was 332+/-196 pg/ml (p<0.01). Mean value of PCT in serum was 80+/-16 ng/ml (p<0.01). The investigated parameters do not permit the identifying of cases with neurologic complications. The increased correlation coefficient between cytokines in serum and in CSF suggests the damage of the blood-brain barrier. The raise of PCT in serum, induced by TNF-alpha and IL-6, is an argument of the severity of sepsis. PMID- 15529590 TI - Limited Wegener's disease presenting as pharyngolaryngeal tumor. AB - Wegener's disease (WD) which is mostly a systemic illness rarely presents as isolated, monoorganic, limited disease. Limited pharyngolaryngeal WD is thus a very rare occurrence. We report the case of a 29 years old man who developed a pharyngolaryngeal tumor with clinically benign evolution, histologically showing granulomatous inflammation and small vessel vasculitis, with no signs of: tuberculosis, sarcoidosis, fungal disease, Hodgkin's disease or foreign body aspiration. p-ANCA's were positive. He was considered a limited form of WD and treated with moderate doses of corticoids and cotrimoxazole. One month later, the lesion diminished significantly. The finding of a pharyngolaryngeal tumor with granulomatous inflammation and vasculitis, in the context of p-ANCA positivity and without any evidence for another systemic granulomatous disease, suggested the diagnosis of limited WD. The response to treatment favoured this presumption. Limited pharyngolaryngeal WD is a rare disease, with a potential for life threatening (even fatal) complications. It should be recognized early and treated promptly. Remissions can be achieved (even without the use of cyclophosphamide). PMID- 15529591 TI - Up to date in pharmacological treatment of dilated cardiomyopathy. AB - Dilated cardiomyopathy is a syndrome characterized by dilatation and impaired function of one or both ventricles and represents the most common heart failure entity requiring heart transplantation. The treatment we currently use does not directly influence specific underlying pathomechanisms of this disorder, and several attempts are being made in order to introduce more specific therapies. We summarized the most important pharmacological strategies that are being evaluated for the treatment of dilated cardiomyopathy and pointed out the novel therapeutic approaches concerning heart failure secondary to this clinical condition. PMID- 15529592 TI - Aldosterone and cardiovascular pathology: from bench to bedside. AB - Thirty years ago, aldosterone was an unidimensional hormone considered responsible for salt and water homeostasis. The current role of aldosterone in pathology is very complex. Recognition of mineralocorticoid receptors throughout the body has raised the importance of aldosterone in understanding the pathophysiology of previously unknown cardiovascular effects. This came from the first documentation of aldosterone receptors outside the kidney, making it not only an endocrine hormone, but a paracrine one, too. Aldosterone is contemporary appreciated to play an important role as a cardiac and vascular risk hormone. Aldosterone-mediated actions include the expression of several genes: collagen genes; genes controlling tissue growth factors and plasminogen activator inhibitor type 1; genes mediating inflammation. Aldosterone receptor blocking agents improve cardiovascular structure, function, and prognosis. They appear to provide additive benefit when used in conjunction with either an ACE inhibitor or an angiotensin receptor blocker. PMID- 15529593 TI - New bioactive angiotensins formation pathways and functional involvements. AB - After a brief review of the actual knowledge concerning the circulating and tissue Renin-Angiotensin System (RAS) as a unitary hormonal system, the cognitive acquisitions regarding the formation and action mechanisms of the new biologically active angiotensins will be presented. The review of the enzymatic pathways for their synthesis and inactivation, as metabolism products of angiotensin II (1-8), will be followed by the presentation of the main physio pharmacological actions of angiotensin III (2-8), angiotensin IV (3-8) and angiotensin (1-7). The functional involvements of the cerebral angiotensin IV in what concerns its possible participation in the normal neurochemical processes of memory and in the neurodegenerative processes of Alzheimer disease will be exposed, together with the vasodilating effects of angiotensin (1-7) as counteracting factor for the constricting effects of angiotensin II. The data concerning the bioactive fragments of angiotensin II will be accompanied by those regarding its implication in the cardiovascular modeling and the induction of oxidative stress, inflammation, atherogenesis, etc. In their turn, personal researches bring new experimental evidences in favor of interactions between angiotensin (1-7) and angiotensin II within the rat thoracic aorta. Biphasic, dose-dependent effects were observed for angiotensin (1-7), induced both through nitric oxide, kinins and prostaglandin release for counteracting the vasoconstricting effects of angiotensin II and the modulation of its own vasodilator action. PMID- 15529594 TI - Cholangiocarcinoma: risk factors, diagnosis and management. AB - Cholangiocarcinomas are malignant tumors of the intra- or extrahepatic biliary tract. An increasing incidence of cholangiocarcinomas has been documented. This increase might be only apparent, due to the progress in investigation and changes in tumor codification. The major clinical sign of cholangiocarcinomas is obstructive jaundice, which is persistent and progressive. Biological tumor markers are nonspecific: an increased serum level of carcinoembryonic antigen is relevant when associated with an increased level of CA 19-9 or CA-125. K-ras mutation and aberrant expression of p53 are present in one third of intrahepatic cholangiocarcinomas. The firstline imaging investigation is ultrasonography, which always detects dilatation of the bile ducts, but more rarely the tumor itself. Classically, endoscopic retrograde cholangiopancreatography (ERCP), the "gold standard" investigation in case of obstructive jaundice, has been performed following ultrasonography. The actual recommendations, based on grade B and C evidences, are to start investigations with ultrasonography and to continue with noninvasive methods: MRI/MRCP or spiral CT, whenever a malignant obstructive jaundice is suspected. Invasive cholangiography (ERCP, PTC) should be reserved for tissue diagnosis or therapeutic decompression when cholangitis is present, or for stent insertion in unresectable tumors. If MRI, CT or cholangiography do not exclude resectability, hepatic arteriography and portal vein evaluation should be performed preoperatively. All patients who do not have unequivocal cholangiographic and angiographic signs of unresectability should undergo surgery, in order to benefit of a possible tumor resection. The radical surgical procedures relieve the obstruction and jaundice by resecting the tumor. The palliative (surgical or endoscopic) procedures cure the jaundice, but do not remove the tumor. Prognosis of cholangiocarcinomas is dismal, although five-year survival rates for these tumors have improved due progress in surgery and adjuvant oncological therapy. PMID- 15529595 TI - The immunopathogenesis of viral hepatitis. AB - The outcome of the HBV and HCV infections is influenced by the status of the immune system. Both of the viruses are believed not to be directly cythopathic, and the host immune response is responsible for the viral clearance and cellular injury. An energic response of the natural immunity allows the control of the virus. Rapid development of specific neutralizing antibodies is essential for the clearance of VHB. The chronic phase of the disease is often associated with abnormal and weak T-cell response and viral persistence. The pathogenesis of the HCV infection is not yet completely understood. HCV persists in patients without any apparent evidence of immune deficits depending on virus or host-related factors. The recent studies revealed that both cellular and humoral immunity appear to be active, despite the progression of the disease. PMID- 15529596 TI - Extrahepatic manifestations in hepatitis C virus infection. AB - The authors present a revision of the literature and also their own experience concerning extrahepatic manifestations in hepatitis C virus infection. A special attention receives the lymphotropism of HCV with mixed cryoglobulinemia and leukocytoclastic vasculitis, lichen planus and porphyria cutanea tarda. PMID- 15529597 TI - Current aspects in etiopathogenesis and diagnosis of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a frequent disease that represents a major cause of disability and mortality in all countries, resulting in an important economic and social burden. The disease results from interaction between individual risk factors (like enzymatic deficiencies) and environmental exposures to noxious agents, like cigarette smoking, occupational dusts, air pollution and infections in childhood. The main mechanisms that may contribute to airflow limitation in COPD are fixed narrowing of small airways, emphysema and luminal obstruction with mucus secretions. COPD is characterised by a chronic inflammatory process in the pulmonary tissue, with a pattern different from bronchial asthma, associated with extrapulmonary effects and is considered now a complex, systemic disease. Optimal therapeutic targeting of COPD depends on a clear understanding of the precise mechanisms of these complex processes and on early and correct evaluation of disease severity. PMID- 15529598 TI - C-reactive protein, prothrombotic imbalance and endothelial dysfunction in acute coronary syndromes without ST elevation. AB - Inflammation is considered a crucial step in the pathogenesis of acute coronary syndromes (ACS). C-reactive protein (CRP) is proposed to be included in risk stratification of ACS patients. However, it is not yet known if CRP is only a risk marker, or merely a risk factor in the development of ACS. Our study looked at the links between inflammation and the prothrombotic factors present in patients with ACS without ST elevation. MATERIAL AND METHODS: 86 patients (pts), 46 men (53.4%), mean age = 58.2+/-12.4 years-old, with acute coronary syndromes (unstable angina and NSTEMI). The following parameters were measured in all pts on admission: CRP, fibrinogen, blood white cell count, and coagulation parameters: coagulation factor V and VIII, von Willebrand factor (vWf), antithrombin III (AT III), D-dimers (DD), C and S proteins. RESULTS: Mean CRP in the study group was 22.42+/-19.81 mg/dl (limits 1.40-88.8 mg/dl). We worked with quartiles of CRP plasmatic levels, in order to see how magnitude of inflammation correlates with different coagulation and fibrinolysis parameters. When comparing the 1st with the 4th CRP quartiles, we noted that important inflammation (4th quartile) was associated with higher factor von Willebrand (141.3 vs 108.9%, p<0.05), factor 5 (127.5% vs 88%, p<0.01), factor 8 (121.5 vs 117.1%, p=0.04), lower AT III (101.6 vs 118.2%, p<0.05), lower protein C and S. The associations did not keep for PAI-I or D-dimers, which might be associated with the lack of sensibility of fibrinolysis markers in the early period after thrombosis. CONCLUSIONS: Inflammation, as quantified by CRP, appears to be associated with a significant prothrombotic status and endothelial dysfunction (as reflected by high von Willebrand factor). PMID- 15529599 TI - The effectiveness and tolerance of betablockers in elderly patients (> or = 70 years) with heart failure. AB - The beta blockers (BB) being less prescribed in elderly patients (P) with heart failure (HF), the aim of this study was to assess the effectiveness and tolerance of BB (meteoprolol, bisoprolol or carvedilol) given on the top of the conventional therapy in HF due to LV systolic dysfunction in P > or = 70 year (n=57, group 1) and < 70 year (n=101, group 2). Differences in baseline clinical characteristics between the 2 groups were not significant. The BB doses given in group 1 P was lower but the difference was significant for bisoprolol only. Intolerance to BB imposing withdrawal occurred in 12% of group 1 P and in 10% group 2 P (p>0.05). Symptomatic improvement expressed as a significant decreases in NYHA class was observed in both groups. Readmission for worsening HF was needed in 42% vs. 39% while 1 year mortality rate was 11.4% vs. 10.4% in group 1 and 2 P respectively (p>0.05). CONCLUSION: BB are tolerated and seem to be effective in most elderly P with HF. Therefore, BB should be tried in all HF P without contraindication irrespective of age. PMID- 15529600 TI - Relationship between serum gamma-glutamyl transferase and pulse pressure in apparently healthy middle-aged subjects. AB - Higher values of pulse pressure (PP)--the difference between systolic blood pressure (SBP) and diastolic blood pressure (DBP)--are associated with an increased cardiovascular risk; SBP and DBP values are correlated with serum levels of gamma-glutamyl transferase (GGT), both in drinkers and non-drinkers subjects. Moreover, some studies showed that high levels of serum GGT might be related with an increased cardiovascular risk. However, few studies analyzed the relationship between GGT and PP. In 72 subjects (18 male; 54 women), middle-aged (mean age: 36.63 yrs; limits: 24-54 yrs), apparently healthy, we performed the measurements of: weight, height (from which we calculated the body mass index [BMI]), SBP, DBP, PP, standard biochemical workup, including GGT (UI/l). RESULTS: PP values are correlated with SBP (r=0.74, p<0.001), but not with DBP (r=0.15, p=NS) values. GGT is significantly correlated with SBP (r=0.28, p=0.01), DBP (r=0.29, p=0.01), and BMI (r=0.41, p<0.001). Mean values of PP are significantly higher in those with a GGT>20 than those with GGT<20 (48.69 vs. 43.85 mmHg, p<0.05); similarly, mean values of SBP (131.66 vs. 116.81 mmHg, p=0.0004) and DBP (82.59 vs. 73.52 mmHg, p=0.0008) are higher in the group with GGT>20. We observed an ascending trend of the mean value of GGT in the different groups of PP; therefore, mean value of GGT was 19.21, 22.65, 23.18 in the PP<40, PP=40-60, respectively PP>60 groups. However, the correlation between PP and GGT was not significant, both in univariate analysis (r=0.14, p=NS) and after adjusting for SBP, DBP, BMI (r=0.12, p=NS). When we analyzed subgroups, the relationship between GGT and PP was more evident in women and in subjects with increased values of BMI. CONCLUSIONS: GGT values are significantly correlated with SBP and DBP, but not with PP, in middle-aged, apparently healthy subjects; even though the highest values of GGT are associated with an increased PP value, further studies would be necessary in order to explain this link. PMID- 15529601 TI - Inflammatory response after dexamethasone eluting coronary stent implantation. AB - C-reactive protein (CRP) elevation after coronary stent implantation is a predictor for recurrence. We prospectively evaluated the inflammatory response after dexamethasone eluting stent implantation in a native coronary artery by serial measurement of plasma level C-reactive protein. METHODS: We investigated patients undergoing primary successful implantation of a single dexamethasone stent (Dexamet, Abbott Vascular Devices, Redwood City, CA, USA) in a native coronary artery. We analyzed plasma concentrations of CRP by immunoturbidimetric assay before stent implantation and 12, 24, 48 and 72 hours after the procedure. Patients on anti-inflammatory drugs or with evidences of inflammatory conditions were excluded. RESULTS: Seventeen patients (mean age 62+/-9 years, 12 males) were enrolled. The presentation was unstable angina in 13 patients and stable angina in 4 patients. Eighteen stents were implanted as follows: 16 type B lesions (88%), 1 type C lesions (6%) and 1 type A lesion (6%), located in LM in 2 patients (11%), LAD in 8 (44%), LCX/OM in 7 (39%), and RCA in 1 patient (6%). The mean CRP increased from 5.6+/-2.2 mg/l at baseline to a maximum of 6.7+/-2.1 mg/l and than decreased to 5.0+/-1.2 mg/l at 72 hours. At 72 hours plasma concentration of CRP was lower than baseline in 11 patients (65%) and higher in 6 (35%). CONCLUSIONS: Inflammatory response to dexamethasone stent implantation in a native coronary artery is low and peaks at 24 hours. At 72 hours after stent implantation, mean CRP decreased comparing with baseline, CRP becoming lower in 65% patients. Using the stent itself as a platform for drug delivery may be an opportunity to modulate the inflammatory response to coronary stent implantation. PMID- 15529602 TI - Echocardiographic evidences of increased left ventricular pressure and atrial dilatation in patients with drug-resistant paroxystic atrial fibrillation and structurally normal heart. AB - Global myocardial index (GMI) has been significantly related to left ventricular filling pressure. We hypothesized that GMI and echographic indicators of atrial dilatation were significantly different in pAF pts compared to normals. METHODS: 39 patients (pts) without structural heart disease, aged 52+/-10 years with pAF were compared to 36 control-matched pts aged 48+/-16 years. Following parameters were assessed: P-wave duration (Pd), GMI, left atrial dimensions (LAd=M-mode, parasternal, LAt and LAI are measurements of short and long-axis apical four chamber view), surface (LAs), volume (LAv), total ejection fraction (LA EF), right atrial dimension (RAd) and surface (RAs), total atrial surface (TAs=LAs+RAs). LAv was calculated using ellipse formula pi/6 (LAdxLAlxLAt). RESULTS: There was no difference between the 2 groups concerning Pd (p=0.1), LA EF (p=0.23), LAd (p=0.08) and LAt (p=0.06) while the rest of the parameters were significantly higher in pAF pts: GMI: 0.5+/-0.17 vs 0.36+/-0.06 (p=0.001); LAI: 5.4+/-0.5 vs 4.5+/-0.3 cm2 (p=0.001); LAs was founded increased in pAF pts (20.6+/-5.7 vs 16.3+/-2.1 cm2, p=0.001); TAs: 40.6+/-6.9 vs 30.6+/-5.1 cm2, p=0.0001; LAv: 51.6+/-10.4 vs 37.2+/-9.3 ml (p=0.0001). CONCLUSIONS: Although apparently without structural heart disease, pAF pts presented evidence of increased filling pressions in left ventricle and indirect markers of atrial stretch. The role of increased intra-atrial pressure in pts pFA and predictive value of these parameters need to be evaluated in a larger number of patients. PMID- 15529603 TI - Baroreceptor sensitivity assessed with the finger pulse wave alpha index, light reflection rheography and ambulatory blood pressure monitoring at two minutes. PMID- 15529604 TI - The effect of simvastatin associated with ranitidine and alcohol upon serum lipids. AB - Statins are highly effective in lowering TC and LDL-C but the increasing of HDL-C is modest and the cost of long-term treatment is too high. On the other hand it is known that the association of H2 receptor blockers increases the plasmatic level of statins, and moderate alcohol consumption increases HDL-C. The purpose of the study was to compare the effect of statins alone and in association with either H2 receptor blockers or alcohol upon serum lipids in dyslipidemic patients. Three groups of dyslipidemic patients were studied, each of them consisting of 20 patients. The group A was treated with simvastatin 20 mg/day, the group B with simvastatin 20 mg/day + ranitidine 150 mg/day and the group C with simvastatin 20 mg/day + 30 g alcohol/day. Before and after two weeks treatment TC, LDL-C, HDL-C and TG were determined in each patient. For the group A patients the studied lipid fractions were modified into the limits described by the literature; for the group B patients, the decreasing of LDL-C was significantly, higher than for A group; for the group C patients, the incresing of HDL-C was significantly, higher than for A group, after two week alcohol consumption. In conclusions, the association of H2 receptor blockers increases the effect of statins upon dyslipidemia and the association of statins with moderate alcohol consumption increases the effect of statins upon HDL-C but the clinical implication has to be established. PMID- 15529605 TI - Intima-media thickness of common carotid artery, carotid atherosclerosis and subtypes of ischemic cerebral disease. AB - It has been shown that common carotid intima-media thickness (CCA-IMT) is a useful outcome measure in clinical studies and intervention trials reflecting early stage of atherosclerosis and cardiovascular risk. The present study examined the association between CCA-IMT and incident ischemic stroke and its subtypes in 75 cases and 21 controls. Cases with ischemic brain infarctions (IBI) were consecutively recruited and classified into subtypes by CT and Bamford's classification (the size and site of infarct) like total anterior circulation infarcts (TACIs), partial anterior circulation infarcts (PACIs), posterior circulation infarcts (POCIs) or lacunar infarcts (LACIs). Controls were recruited among individuals hospitalized at the same institutions and matched for age and sex. Patients and control subjects underwent B-mode ultrasonographic measurements of IMT of the distal wall of both common carotid arteries. Of 75 patients with an acute ischemic stroke, 10 (14%) were classified as TACIs, 34 (45%) with PACIs, 12 (16%) with POCIs and 19 (25%) with LACIs. Mean CCA-IMT was higher in cases (1.03+/-0.18 mm) than in controls (0.85+/-0.18 mm; p<0.0001). The difference in CCA-IMT between cases and controls was significant and the relation between CCA IMT and IBI remained after adjustment for main cardiovascular risk factors. Regarding the subtypes of IBI, IMT values were significantly higher in patients with TACIs and PACIs versus those with LACIs and POCIs. An increased CCA-IMT was associated with the all subtypes IBI and was significantly higher in those with anterior circulation infarcts versus posterior circulation and lacunar infarcts. PMID- 15529607 TI - The role of octreotide on renal function in patients with advanced cirrhosis. AB - Advanced cirrhosis is characterized by arterial vasodilatation and the reduced arterial response to vasoconstrictors. Both of these are related to an increase of endothelial (prostacyclin and nitric oxide) and nonendothelial vasodilatators (glucagon) level. Experimental study had shown that the responsiveness to vasoconstrictors of the mesentery artery may be normalized by the inhibition of glucagon or nitric oxide. The aim of our study was to assess the effects on renal hemodynamics by the administration of octreotide, the synthetic somatostatin analog, an inhibitor of the release of endogenous vasodilatators. METHODS: We studied forty-six patients admitted at the University Hospital in Bucharest for advanced cirrhosis between 2000-2002. The patients aged (35-62) were divided in two groups: Group I--23 patients received intravenous octreotide by infusion of 50 microg/hour for three days; Group II--23 patients received placebo. Exclusion criteria were: A recent gastrointestinal bleeding; Hepatic encephalopathy; Serum creatinine > 1.5 mg/dl; Ultrasonographic evidence of renal disease or urinary tract obstruction. STUDY PROTOCOL: Discontinuation of pharmacological treatment with beta-blockers, nitrates and angiotensin converting enzyme inhibitors 7 days before the study; Sodium restricted diet (35 mEq/day) 2 weeks before and during the study. All the patients underwent on day 0 and 3 the following; Routine laboratory tests, Creatinine clearance for glomerular filtration rate (GRF); Urinary sodium excretion (24 hours), Water diuresis, Lithium clearance, Plasma renin activity, Plasma aldosterone concentration, Medium blood pressure, Cardiac output (by ecocardiography). CONCLUSIONS: Octreotide in a short infusion treatment induces an inhibitory effect on renin aldosterone secretion which may be responsible for the benefical effects on sodium excretion. The significant increase of MAP may be the result of an inhibition of vasodilatatory substances or an increased arterial responsiveness to vasoconstrictors. The significant effect on renal function in patients with advanced cirrhosis can be a promising therapeutic perspective in the treatment of hepatorenal syndrome. PMID- 15529606 TI - Therapeutic efficiency of continuous renal replacement therapy--experience of a single Romanian center. AB - We have studied 24 patients with acute renal failure (ARF) or acutization of chronic renal failure (CRF) caused by a variety of different disorders. A rapid decline in renal excretory function, fluid, electrolyte, and divalent ion disturbances determined us to initiate continuous renal replacement: continuous venovenous hemofiltration (CVVHF)--28 procedures in 15 patients and continuous venovenous hemodiafiltration (CVVHDF)--11 procedures in 11 patients. Mean duration of CVVHF was 21.6+/-6.9 h, with a mean blood flow rate of 116.9+/-16.4 ml/min and an ultrafiltration rate of 6.4+/-4.6 ml/min. Serum creatinine level decreased from 12.6 to 8.3 mg% and the concentration of urea from 237 to 166 mg%. Mean duration of hemodiafiltration was 24+/-8.5 h, with mean blood flow rate of 134+/-15.2 ml/min, mean dialysate flow of 35+/-7 ml/min and mean ultrafiltration rate of 5.6+/-2.1 ml/min. Serum creatinine level decreased from 11.6 to 6.36 mg% and the concentration of urea from 236 to 137 mg%. Survival rate was 79.2% (19/24 patients). Complete recover of renal function was achieved in 5 patients, partial recover in 5 patients and 9 patients were included in chronic dialysis. In 8/24 (30%) patients we encountered complications, such as hypotension in 2 cases, bleeding disorders in 5 cases or diselectrolithemias in 2 cases. As a consequence, continuous renal replacement therapy is efficient, having an acceptable rate of adverse effects in patients with ARF or acutization of CRF. PMID- 15529608 TI - The prevalence of anti-Helicobacter pylori antibodies in the patients with ischemic heart disease. AB - The authors proposed to assess the prevalence of anti-Hp antibodies (IgG class) in patients with ischemic heart disease. The study included 93 patients distributed into two groups: a) group A--61 patients with ischemic heart disease; b) group B--32 controls. The Hp infection diagnosis was established by determination of the Hp antibodies in the peripheral blood. Hp infection was correlated with the following factors: (a) smoking, alcohol consumption, antiinflammatory drugs intake and water source; (b) biological parameters (fibrinogen, alpha1-globulins, cholesterol, triglycerides, thrombocyte count). In the patients with ischemic heart disease the prevalence of the Hp infection was 78.3%, and in controls 56.2% (p<0.05). The patients consuming alcohol and smoking had a significantly higher prevalence than controls. Antiinflammatory drugs and water source did not influence the prevalence of the Hp infection. Among the biological parameters, fibrinogen and alpha1 globulins were significantly increased in the Hp-positive patients with ischemic heart disease. In conclusion, the serum prevalence of anti-Hp antibodies was higher in the patients with ischemic heart disease, smokers and alcohol consumers than in health individuals. PMID- 15529609 TI - The effect of two antidiarrhoeal drugs on the psychosocial adjustment to illness in chronic functional diarrhoea. AB - BACKGROUND: Chronic functional diarrhea is an embarrassing symptom, causing the impairment of the quality of life. AIM: We looked for the effect of two antidiarrheal drugs, diosmectite and loperamide on the psychosocial adjustment to illness, a predictor for the health-related quality of life, in chronic functional diarrhea. METHODS: Prospective controlled randomized trial including 50 consecutive patients with chronic functional diarrhea. They received either diosmectite 3 g b.i.d (n=25, 11M/14F, age 43+/-12 years) or loperamide 2 mg b.i.d. (n=25, 12M/13F, 47+/-11 years) for 2 weeks. Diarrhea and four other digestive symptoms were evaluated on a 0-16 scale, as well as patient's satisfaction (scale 1-10) and physician's satisfaction (scale 0-4). Psychosocial adjustment to illness was assessed with the PAIS-SR scale. RESULTS: Symptom score for diarrhea was reduced from 10.5+/-5.7 to 2.6+/-1.2 (mean+/-SD), p<0.001 by diosmectite and from 8.5+/-4.2 to 1.2+/-0.6, p<0.0001 by loperamide. Diosmectite had a stronger effect than loperamide on accompanying pain (p<0.05) and bloating (p<0.01), but not on nausea, anorexia and emesis. The overall ratings of the patient's satisfaction (8.7+/-0.9 vs. 8.2+/-1.1, NS) and physician's satisfaction (3.2+/-0.3 vs. 3.1+/-0.4, NS) were similar. Psychological adjustment to illness was not changed for following items: health care orientation and domestic environment. Diosmectite improved more than loperamide the psychological distress (from 12.7+/-3.8 to 7.7+/-2.1 vs. 11.8+/-3.3 to 9.05+/-2.2, p<0.05), vocational environment (from 3.2+/-1.1 to 2.2+/-1.2 vs. 3.3+/-1.2 to 3.4+/-1.5, p<0.05) and sexual relationship scores (from 6.4+/-2.2 to 3.0+/-1.5 vs. 7.2+/-2.2 to 5.5+/ 2.1, p<0.05). Both drugs similarly improved the items: family relationship and social environment. CONCLUSIONS: Diosmectite and loperamide are both potent drugs for chronic functional diarrhea. Diosmectite has better effects on associated symptoms and some items of psychosocial adjustment to illness (distress, sexual activity and vocational environment). PMID- 15529610 TI - Evaluation of a Romanian version of the Bath Ankylosing Spondylitis Functional Index (BASFI) in patients with spondylarthropathies. AB - OBJECTIVE: To develop a Romanian version of the Bath Ankylosing Spondylitis Functional Index (BASFI) and to determine its metric properties in patients with spondylarthropathies (SpA). METHODS: A Romanian version of BASFI was obtained after translation, back-translation and pretesting. Altogether 41 SpA patients fulfilling the ESSG criteria were included, 34 of them satisfying the modified New York criteria for ankylosing spondylitis. BASFI was completed by the patients along with assessment of clinical and disease activity variables by both the patient and physician. Scalability, internal consistency, reliability and construct validity of the Romanian BASFI were assessed. RESULTS: Mean time to complete the questionnaire was 2 minutes. Scalability (0.2-9.7, 95% of the scale), internal consistency (Cronbach's alpha = 0.93), reliability (intraclass correlation coefficient = 0.82) and construct validity (correlations (absolute value(r) = 0.37-0.59) with spinal pain, nocturnal pain, patient's and physician's global assessment, fingers-to-floor distance, occiput-to-wall distance, modified Schober test, intermalleolar distance, and Dougados articular index) were all very satisfactory. CONCLUSIONS: Translation and cultural differences did not alter the well-known value of BASFI. Therefore, the Romanian version of BASFI is reproducible and valid for the assessment of patients with spondylarthropathies. PMID- 15529611 TI - Aspects of nailfold capillaroscopy in children and young people with rheumatic disease. AB - Nailfold capillaroscopy was performed in children and young people with rheumatic disease (42 patients with a variety of childhood rheumatic diseases) and 30 normal control subjects. Distinctive morphologic abnormalities with capillary enlargement, megacapillary and dropout of surrounding structures were noted in two groups: patients with childhood dermatomyositis (DM) and with scleroderma (SD). Capillary abnormalities were found in all 4 patients affected by systemic disease and in none of 7 patients with localized scleroderma. Some minor atypical capillary morphology were observed in normal individuals (37%) but the enlarged capillaries, especially megacapillaries were not found. PMID- 15529612 TI - Wavelets as methods for ECG signal processing. PMID- 15529613 TI - Treatment of iron deficiency anemia with Ferro-Folgamma. AB - Iron deficiency anemia is a hypochromic anemia in which hemoglobin poor synthesis is due to a decrease in the amount of iron in the body. The decrease of iron quantity has many causes: insufficient intake of aliments rich in iron (meat, viscera, green vegetables), increased necessities during growth period, pregnancy, erythrocytes hyperregeneration, high-performance sportsmen, increased loss by digestive way, genito-urinary way, respiratory, hemorrhagic syndromes. Clinically, symptoms and signs specific to all types of anemia and those specific to lack of iron occur besides the symptoms and signs of the underlying disease: atrophic glositis, angular stomatitis, sideropenic dysphagia, pica, skin and nails changes. Laboratory investigations useful for diagnosis are: microcytic, hypochromic anemia, decreased serum iron level, total capacity of iron binding increased, medullar iron store absent, good response to iron therapy. Ferro Folgamma is one of the most indicated medicines in iron deficiency anemia. Due to its components this medicine has many indications: insufficient alimentary intake concerning iron, folic acid, B12 vitamin, vegetarian alimentation, increased needs during growth period, iron deficiency anaemia secondary to chronic hemorrhages, malnutrition, anemias associated with chronic alcohol intake, preventive treatment of iron deficiency anemia and megaloblastic anemia during pregnancy and lactation. PMID- 15529614 TI - Metabolic syndrome--practical approach. AB - The diagnosis of metabolic syndrome is based on identification of the following parameters: abdominal obesity, triglycerides, HDL-cholesterol, blood pressure, fasting glycemia, as recommended by ATP III. In order to simplify the clinical practice, at least two parameters should be screened for. The most frequent couple, easy to be determined in practice, is hypertensive waist, followed by hypertriglyceridemic waist, hypertensive dyslipidemia, dysglycemic dyslipidemia and hypertensive dysglycemia. Based on these couples the next step would be to identify the triads that diagnose the metabolic syndrome. A global assessment of cardiovascular risk should be made. Suggested method is to apply the Framingham Score. Therapeutic intervention is structured according to levels of cardiovascular risk. Clinical management is structured on THEME Programs (therapy, education, monitoring, evaluation), applied to all risk factors. PMID- 15529615 TI - The metabolic syndrome--a multifaced disease. AB - The metabolic syndrome (MetS) is a huge public health problem worldwide, being one of the major causes of cardiovascular disease, responsible for a growing number of premature deaths throughout the world. MetS includes a cluster of anomalies, such as: abdominal obesity, insulin resistance, hyperinsulinemia, hypertension, type 2 diabetes mellitus or glucose intolerance, hypertriglyceridemia etc. The number of people with MetS increases with age, affecting more than 40% of people in their 60s and 70s. About 30% of European people over 50 have MetS. Some experts estimate that as many as two thirds of Americans may be suffering from MetS. The exact cause of MetS is not known: genetics play a minor role, acquired in-utero factors also play a role (prenatal malnutrition, toxin exposure, exposure to high levels of maternal cortisol). For most people, the MetS results primarily from lifestyle factors, such as: chronic stress, inadequate exercise. The MetS can be avoided and reversed in most cases. Weight loss is both a treatment and goal for MetS patients. Moderate weight loss, in the range of 5-10% of body weight, can help restore body's ability to recognize insulin and greatly reduce the chance that the syndrome will evolve into a more serious illness. In most people weight loss will lower blood pressure and improve triglyceride levels. Increased activity alone can improve insulin levels. Physical activity result in a weight loss, improved blood pressure, improved cholesterol and triglyceride level and reduced risk of developing diabetes. It is also important to treat: hyperlipidemia, hypertension, prothrombotic state. PMID- 15529616 TI - Metabolic syndrome--new insights into a growing entity. AB - Metabolic syndrome is a cluster of cardiovascular risk factors. Pathogenesis of metabolic syndrome implies 3 potential etiological mechanisms: obesity and adipose tissue disorders, insulin resistance, and a constellation of independent factors. Clinical recognition of the metabolic syndrome is based on finding several well-recognized signs in clinical practice: abdominal obesity, elevated triglycerides, reduced HDL cholesterol, raised blood pressure, and elevated plasma glucose. In addition, other components commonly aggregate with the major components: elevated apolipoprotein B, small LDL particles, insulin resistance and hyperinsulinemia, impaired glucose tolerance (IGT), elevated C-reactive protein (CRP), and variation in coagulation factors (plasminogen activator inhibitor [PAI]-I and fibrinogen). Cardiovascular disease (CVD) is the primary clinical outcome of metabolic syndrome. Additionally, risk for type 2 diabetes is higher. Diabetes is itself a major risk factor for CVD. ATP III criteria for diagnosis of metabolic syndrome provide a practical tool to identify patients at increased risk for CVD. World Health Organization (WHO) and American Association of Clinical Endocrinologists (AACE) criteria require further oral glucose testing if IFG and diabetes are absent. IGT on OGTT denotes greater risk for diabetes than does metabolic syndrome without elevated fasting glucose. PMID- 15529617 TI - Heart failure and diabetes. AB - Diabetes substantially increases the risk of heart failure both in men and women, being included in the Stage A classification of heart failure by the American Societies of Cardiology. The main etiological factors contributing to heart failure in diabetes are coronary artery disease, systemic hypertension and diabetic cardiomyopathy, the latter being invoked in case of heart failure where the first two factors are missing. Renal insufficiency and obesity may also play a role. The diagnosis will follow the same steps as in non-diabetic subjects: careful and periodic assessment for signs and symptoms of heart failure in all diabetic patients, echocardiography to assess the systolic and diastolic function of the left ventricle, and B-type natriuretic peptide level (as a marker of left ventricular dysfunction). The therapeutic approach will include non pharmacological measures and pharmacological treatment. Patients with diabetes and heart failure benefit of the same drugs as non-diabetic subjects, including beta-blockers, which should not be avoided in patients with diabetes. The antihyperglycemic agents that should not be used in patients with heart failure are biguanides and thiazolidindiones (pioglitazone can be used in NYHA I and II classes). Approaches that were proven to reduce the risk of heart failure in diabetes are blood pressure and lipid control, treatment with ACE inhibitors in patients with diabetes and other cardiovascular risk factors and improvement of the glycemic control. PMID- 15529618 TI - Renin-Angiotensin-Aldosterone system inhibition in prevention of diabetes mellitus. AB - The number of people with diabetes grows worldwide. The complications resulting from this disease are a significant cause of morbidity and mortality. World Health Organization estimates that, while in the year 2000 the number of people with diabetes was about 177 million, by 2025, this will increase to at least 300 million. The diabetes epidemic, without primary prevention, will continue to grow. Individuals with type 2 diabetes are at a significantly higher risk for coronary heart disease, peripheral vascular disease, and stroke, and they have a greater probability of having hypertension, dyslipidemia, and obesity. A number of clinical trials provide evidences that RAAS inhibition could be helpful at preventing new onset of type 2 diabetes mellitus. Pharmacologic treatment that antagonize the renin-angiotensin system (RAS) provide more benefits, not only in patients after myocardial infarction and in congestive heart failure, but also in persons with hypertension and type 2 diabetes mellitus. PMID- 15529619 TI - Quality of life evaluation in diabetes mellitus. PMID- 15529620 TI - Treatment of diabetic polyneuropathy with alpha-lipoic acid is evidence based. AB - Searching for more efficient treatments for diabetic neuropathy continues to be a task for researchers and for practitioners also. The reasons are: the poor quality of life and the risk for ulcerations/amputations induced by peripheral diabetic neuropathy (PDP) (the most frequent manifestation of diabetic neuropathy) and the risk for sudden death and other manifestations attributed to autonomic neuropathy. There is a paucity of evidence based interventions for PDP. In this situation, the recent evidence (randomized, placebo controlled studies) regarding the efficacy of alpha-lipoic acid, a powerful antioxidant, are bringing new hopes. PMID- 15529621 TI - Atypical antipsychotics and diabetes mellitus. AB - Numerous findings point towards a higher incidence of diabetes mellitus and cardio-vascular risk factors among patients with schizophrenia compared with the general population. The introduction of atypical antipsychotics, which are much more effective than classic neuroleptics in the treatment of schizophrenia, was associated with an increase of (sometimes severe) high blood sugar and diabetes cases. The mechanisms hypothesized to be involved in the "diabetogenic effect" of atypical antipsychotics are the facilitation of weight gain and the decrease of sensitivity to insulin. There are important differences among antipsychotics as far as both their psychotropic profiles and the magnitude of such so called "diabetogenic" effects are concerned. Usually, risk factors of diabetes precede and predict the development of diabetes mellitus. In the absence of a proven effect on beta pancreatic cells, it appears that atypical antipsychotics facilitate the development of diabetes in individuals at high risk who do not have the capacity to increase their secretion of insulin to a sufficient extent as to compensate the resistance of tissues to the action of insulin. The monitoring of patients under treatment with atypical antipsychotics, in particular of patients with risk factors of diabetes, and the choice of antipsychotics in relation with the particular psycho metabolic features of patients, are the recommended attitude in order to increase the benefit and decrease the risk of treatment. PMID- 15529622 TI - Strong association of insulin gene INS-VNTR polymorphisms with type 1 diabetes in the Romanian population. AB - Type 1 diabetes (T1DM) is a common, chronic disease with autoimmune pathogeny, conditioned by genetic factors. Class II HLA DR and DQ and insulin gene polymorphisms encode for most of the T1DM genetic susceptibility. We have previously shown that class I alleles of the insulin gene INS-VNTR locus are strongly associated with T1DM in the Romanian population. The aim of our study was to confirm the contribution of INS-VNTR to T1DM genetic susceptibility in Romania. For this we typed the insulin gene -23HphI A/T polymorphism (an accurate marker for the INS-VNTR alleles) on 219 Romanian T1DM families using Taqman. Allele transmission to diabetics and unaffected siblings was assessed using the Transmission Disequilibrium Test (TDT). We found a significantly increased transmission of -23HphI A allele to diabetics (78.31% transmission, pTDT = 2.4 e 07) which confirms our previous findings. Combined with the data from the first 204 Romanian T1DM families, the transmission of -23HphI A allele to diabetics is almost 80% (79.78%, pTDT = 2.8 e-15). This percentage indicates the same level of predisposition as for the most diabetogenic HLA's. In conclusion, our results indicate an exceptionally strong association of the class I INS-VNTR alleles with T1DM for the Romanian population. PMID- 15529623 TI - GADA and islet cell antibodies in Romanian children and adolescents with diabetes mellitus. AB - Childhood type 1 diabetes is defined by autoimmunity and insulinopenia. Etiopathogenic definition based on biochemical characteristics has recently replaced the clinical definition based on insulin requirement for treatment. The aim of this study was to describe biochemical and clinical characteristics of children with clinically diagnosed type 1 diabetes, hospitalized at the "Cristian Serban" Center in Buzias. PATIENTS AND METHODS: Fasting C peptide, HbA1c, islet cell autoantibodies (ICA) and antibodies against glutamic acid decarboxylase (GADA) were measured in 278 subjects aged (mean +/- SD; range) 15.1 +/- 4.8 (4 28) years, with a disease duration of 2.1 +/- 0.7 (1.1-3.1) years. GADA and ICA positivity was defined by values higher than the 95th percentile in 99 age matched non-diabetic controls (0.4 units for ICA and 1.4 for GADA). RESULTS: As many as 66.2% of all patients had positive GADA and 10.1% had positive ICA. While 68.7% had at least one positive antibody, only 7.6% had both antibodies positive. As expected, most of the children (79.9%) had fasting C peptide values in the low range (<0.5 ng/ml), but 3 patients (1.1%) had biochemical signs of insulin resistance (C peptide concentrations >3 ng/ml). Two of the three insulin resistant children had positive GADA and one of them had positive ICA, therefore showing "mixed" features of both type 1 (autoimmunity) and type 2 diabetes (insulin resistance). CONCLUSIONS: Childhood diabetes is now acknowledged to be a complex disorder with heterogeneity in its pathogenesis, clinical course and outcomes. While type 1 diabetes is the most frequent form of diabetes among Caucasian children, measurement of diabetes autoantibodies and C peptide is necessary to better define the types of diabetes in youth. PMID- 15529624 TI - The value of basal C peptide and its relationship with pancreatic autoantibodies in young adults with type 2 diabetes mellitus. AB - It is well known that sometimes it is difficult to distinguish between type 1 and 2 diabetes mellitus using clinical criteria, in subjects with disease onset relatively early in adult life. The measurement of C peptide level and of immunological markers may represent important additional tools for establishing the correct diagnosis. The aim of the study is to assess the trend of basal C peptide in patients with clinical diagnosis of type 2 diabetes and to relate it to the type of treatment, the body weight and the positivity for pancreatic autoantibodies. PATIENTS AND METHOD: we studied a group of 268 patients with type 2 diabetes, aged between 30 and 50 years, with a diabetes duration of less than 5 years. In all patients, we measured basal C peptide, islet cell autoantibodies and antibodies against glutamic acid decarboxylase, computed the body mass index and recorded the current antidiabetic treatment. RESULTS: Based on basal C peptide value, diabetic subjects fell under 3 categories: a) low C peptide (<0.58 ng/ml): 7.5%, b) normal C peptide (0.58-2.70 ng/ml): 57.8%, and c) high C peptide (>2.70 ng/ml): 34.7%. Patients with low C peptide were treated more often with insulin, while those in high C peptide group received more often biguanides. A direct correlation between C peptide and body weight was established. Mean C peptide was lower in patients positive for at least one pancreatic autoantibody, compared to those who were negative for antibodies CONCLUSION: Low basal C peptide can be considered criterion for transferring the patients, initially diagnosed as type 2 diabetes, in the type 1 diabetes group. PMID- 15529625 TI - Early detection of atherosclerosis in type 2 diabetic patients by endothelial dysfunction and intima-media thickness. AB - Endothelial dysfunction (ED) is regarded as an early functional marker and intima media thickness (IMT) as an early morphological marker of atherosclerosis. The aim of this study was to evaluate whether peripheral endothelial function and common carotid IMT are impaired in type 2 diabetic patients with inadequate glycemic control. MATERIAL AND METHODS: We examined 20 subjects--10 controls (aged 54.2 +/- 4.37 years) and 10 type 2 diabetic patients (aged 55.6 +/- 3.66 years) with HbA1c level >8.5% (mean 9.25% +/- 0.54%), with an average duration of diabetes of 5.65 +/- 2.34 years, all without clinical evidence of atherosclerosis. B-mode ultrasonography was used to assess both intima-media thickness in the common carotid artery and endothelium function in the brahial artery. In order to evaluate endothelium function brachial artery diameter was measured at rest, during reactive hyperemia, which causes endothelium-dependent vasodilation and after sublingual administration of nitroglycerin (500 microg), which causes endothelium-independent vasodilation. RESULTS: There was no statistical difference between the two groups regarding age. Systolic blood pressure, diastolic blood pressure, total cholesterol were higher and HDL cholesterol was lower in the diabetic group, but the differences didn't reach statistical significance. Triglycerides concentration and BMI were significant greater in the diabetic group (triglycerides-204 +/- 48.5 mg/dl vs. 117 +/- 22.2 mg/dl, p<0.0001; BMI-28.6 +/- 1.28 kg/m2 vs. 26.7 +/- 1.93 kg/m2, p=0.021). Diabetic subjects had significantly impaired flow-mediated vasodilation (FMD) in the brachial artery compared with control group (5.65% +/- 1.42% vs. 7.11% +/- 1.01%, p=0.016). Endothelium-independent vasodilation induced by nitroglycerin did not differ between the two groups (15.4% +/- 2.01% in diabetic subjects vs. 15.7% +/- 2.58% in control subjects, p=0.75). Intima-media thickness was significantly increased in diabetic patients (0.87 +/- 0.07 mm vs. 0.77 +/- 0.06, p=0.0022). CONCLUSION: In diabetic patients with inadequate glycemic control and without clinical evidence of atherosclerosis endothelial function assessed by FMD is significantly impaired and IMT is significantly greater compared to nondiabetic healty subjects. Large clinical trials should evaluate if in clinical practice FMD and IMT are useful in identification of high-risk subjects. PMID- 15529626 TI - The quantification of cardiovascular risk factors in subjects with/without diabetes mellitus. AB - Data from the literature showed an increased cardiovascular risk in patients with type 2 DM comparatively to subjects without DM, sustaining that diabetes mellitus is a cardiovascular disorder. In this context, we aimed to study the prevalence of modifiable cardiovascular risk factors in patients with type 2 DM comparatively to a control group without DM. The results are alarming, despite the great efforts of the members of the caring team. Smoking and alcohol consumption exceeding 30g/day are more frequent habits in patients with diabetes comparatively to controls, especially in men. Hypertension was present in 70.17% of diabetic patients, comparatively to 40.47% in controls; though already diagnosed, in the majority of diabetic patients, the therapeutic goal was achieved in only 25.45% subjects at the time we performed the study. Obesity including abdominal obesity is present from the moment diabetes is diagnosed and it is maintained further, indifferent of the duration of diabetes. Lipid parameters are very increased. So, it is imperative to intensify the therapeutic medical education, regarding each risk factor. PMID- 15529627 TI - Cardiovascular risk factors in Turner syndrome. AB - Turner syndrome is due to haploinsufficiency of X chromosome genes that escape inactivation and associates female phenotype, short stature, gonadal dysgenesis, somatic stigmata, cardiovascular and renal anomalies and a large spectrum of other disorders (autoimmune thyroiditis, osteoporosis, inflammatory bowel disease, chronic liver diseases). The increased mortality in Turner syndrome is primarily a result of its cardiovascular complications. Congenital cardiac anomalies (coarctation of the aorta, bicuspid aortic valve, anomalous venous drainage) are present in 23-40% of patients; there is an increased risk of aortic dilation (42%) and dissection, ischemic heart disease and the risk of hypertension is increased three fold. In addition, insulin resistance may be present in up to 50% of women with Turner syndrome and an atherogenic lipid profile (increased cholesterol, triglycerides) favors the development of coronary artery disease. Our study was aimed to reveal anomalies in Turner syndrome that may increase cardiovascular risk. We studied a group of 62 Turner patients aged 16-67 years (mean age 26.8 years, SD = 11.1 years) comparatively to 62 age matched controls. Glycemia over 100 mg% was found in 11.3% of Turner patients vs 1.6% of controls and cholesterolemia over 200mg% was found in 51.2% of Turner patients vs 14.5% of controls; 24.2% of Turner patients were overweight vs 17.8% of controls and 6.4% were obese vs 4.8% of controls. In the Turner group we found congenital cardiac anomalies in 17.8%, hypertension in 6.5%, renal anomalies 11.3%, and hypothyroidism 29.2%. PMID- 15529628 TI - Continuous glucose monitoring: physiologic and pathophysiologic significance. AB - Diabetes mellitus is a complex disorder of the energy metabolism. In the present paper, we have tried to illustrate the changes in the regulation of blood glucose levels encountered in the two main types of diabetes: Type 2 (T2DM) and Type 1 (T1DM) diabetes mellitus, compared with healthy, non-diabetic subjects. For this we used the MiniMed CGMS (Continuous Glucose Monitoring System) which allows the continuous in vivo blood glucose measurement over a 3-day period. The study group comprised 19 diabetic patients (14 T1DM and 5 T2DM cases) and 4 non-diabetic controls. The recording in normal subjects showed a glycemic variation between 46 and 118 mg/dl, suggesting the existence of a strong and efficient glycemic control mechanism. In T2DM patients, both on diet only or on oral antidiabetic treatment, the oscillation of blood glucose levels was significantly higher compared to that recorded in non-diabetic subjects. In T1DM patients with stable metabolic control blood glucose fluctuations were comparable with those recorded in long-term type 2 diabetic patients but the "mean" values of blood glucose over 72 hours were lower. The CGMS is a valuable tool in the detection of unrecognized hypoglycemic episodes and hyperglycemic postprandial peaks and allows the patient and the health care team to adjust the treatment regimen in order to improve glycemic control. From our point of view, the CGMS could offer valuable information for the knowledge of glycemic regulation in normal people and for the diabetogenic mechanisms in prediabetic IGT and IFG patients. PMID- 15529629 TI - The oxidative stress in the development of diabetes chronic complications in the elderly. AB - The diabetes mellitus occurs as an important disease at elderly people, to whom the micro- and macrovascular complications represent a major cause of morbidity and mortality. Experimental researches of the last years proved that the oxidative stress may be the common mechanisms that intervenes in the occurrence of the diabetes complications as well as in the aging process and is responsible for the increased prevalence of chronic complications at elderly diabetics. Starting from this information, we performed a comparative study where we followed the intensity of the oxidative stress at elderly diabetics as compared to adult diabetics and non-diabetic elderly people. At the same time, we have followed the involvement of oxidative stress in the occurrence of diabetic microangiopathy and atherosclerosis. 155 patients from the 4th Medical Clinic were studied during 2000-2003. These patients were divided into three lots: lot 1: elderly diabetics, lot 2: adult diabetics, lot 3: elderly non-diabetics. At these patients we have followed comparatively the intensity of the oxidant status by determining the plasma malondialdehyde (MDA) and the anti-oxidant status by determining the plasma ceruloplasmin, as well as the correlations of these two parameters with the chronic complications of diabetes. At elderly diabetics there is an increased oxidative stress underlined by an increased plasma level of MDA and ceruloplasmin as compared to the adult diabetics and non-diabetic elderly people and this increased oxidative stress is involved in the development of the chronic complications at this patients. In case of elderly diabetics, the age and the illness may induce the formation of oxygen-derived free radicals with synergic effect in injuring tissues and organs. PMID- 15529630 TI - Rheological changes in diabetic microangiopathy. AB - OBJECTIVES: Evaluation of rheological changes in patients with diabetes mellitus. MATERIAL AND METHOD: Determination of plasma viscosity, erythrocyte deformability, erythrocyte adhesion, fibrinogen and hematocrit in a group of 56 patients with diabetes mellitus, of which 28 with diabetic microangiopathy. The group includes 20 patients with type 1 diabetes mellitus and 36 patients with type 2 diabetes mellitus. RESULTS: In patients with diabetes mellitus with microangiopathy, more important rheological changes are noted than in diabetes mellitus without microangiopathy; thrombocyte adhesion is increased in 25 patients with microangiopathy compared to those without microangiopathy (7), erythrocyte deformability is decreased in 23 patients with microangiopathy and only in 9 patients without microangiopathy, and increased plasma viscosity is found in 26 patients with microangiopathy compared to those without microangiopathy (3). CONCLUSIONS: In the study performed, a direct correlation is found between the decrease of erythrocyte deformability and the severity of diabetic microangiopathy; the decrease is more severe in patients with proliferative retinopathy or clinically manifest nephropathy. The increase of platelet adhesion in patients with diabetic microangiopathy suggests the hypothesis of the role of thrombocyte changes in the pathogenesis of microangiopathic complications of diabetes mellitus. Plasma viscosity contributes to the pathogenesis of diabetic nephropathy and diabetic polyneuropathy, its increase being noted in diabetic patients with microalbuminuria and peripheral neurological involvement. PMID- 15529631 TI - Metabolic changes in Klinefelter syndrome. AB - Klinefelter syndrome is a hypergonadotropic hypogonadism determined by the presence of one or more extra X chromosomes and has an incidence of 1:800 males. It was shown that in Klinefelter syndrome mortality is increased due to diabetes, cardiovascular, respiratory and digestive diseases. Our goal was to assess the presence of metabolic changes which can increase cardiovascular risk. We compared 31 untreated Klinefelter patients aged 19-54 years (mean age 33.84 years, SD 11.79 years) with 31 age matched controls. Hypercholesterolemia was present in 56.67% of Klinefelter patients and correlated with age and the magnitude of androgenic deficit. None of the patients had diabetes mellitus but glycemic values above 100 mg% were present in 16.13% of cases (vs 3.2% in controls). 35.5% of Klinefelter patients and controls were overweight and 16.1% of Klinefelter patients were obese vs 9.7% of controls. PMID- 15529632 TI - Lipid peroxidation and catalase in diabetes mellitus with and without ischemic stroke. AB - Production of reactive oxygen species (ROS) increased in diabetic patients and oxidative damage may contribute to the development of diabetic complications. Malondialdehyde is known as marker of the oxidative damage. Catalase is one of antioxidative factors involved in elimination of ROS In this study, the plasma level of lipid peroxides and plasma catalase activity in 315 patients with diabetes mellitus were assayed. We also included 114 non-diabetic healthy controls whose age, sex were matched to the diabetic patients. The plasma levels of lipid peroxides (LPO) were determined by spectrophotometric method modified by Satoh and Yagi. Lipid peroxidation was estimated by the plasma level of malondialdehyde (MDA). In controls mean value of plasma lipid peroxides was 1.329 +/- 0.118 nmol/ml. In diabetic patients with ischemic stroke MDA level was 2.919 +/- 0.182 nmol/l; p<0.001, and in patients without ischemic stroke the MDA level was 2.329 +/- 0.149 nmol/l; p<0.05; between diabetic patients with ischemic stroke and patients without ischemic stroke p<0.01. The high level of lipid peroxides might induce a self-maintained chronic process which, in time, might lead to the aggravation of the macro- and microangiopathy in diabetes. The plasma levels of catalase were determined by Goth's spectrophotometric method. In 114 healthy persons the mean value of plasma catalase (CAT) activity was 115.3 +/- 14.5 MU/l with less plasma catalase for females (108.7 +/- 12.4 MU/l) than for males (118.9 +/- 16.6 MU/l). Mean value of plasma CAT was (102.4 +/- 12.7 MU/l in patients with ischemic stroke, p<0.001 and 116.3 +/- 18.7 MU/l in patients without ischemic stroke, p<0.05); between diabetic patients with ischemic stroke and patients without ischemic stroke p<0.01. Our results revealed a decrease in plasma CAT activity in patients with diabetes mellitus and ischemic stroke as compared to patients with diabetes mellitus without ischemic stroke. We can conclude that in diabetic patients the decrease in plasma CAT activity is the consequence of oxidative modifications. These results suggest that diabetic patients have significantly increased oxidative damage. PMID- 15529634 TI - Proteic sulphur levels in hair, a possible tool to manage the balance of IDDM. AB - Assays of proteic sulphur levels in the hair of patients with new detected IDDM which were followed up for 6 months, showed a negative correlation between HbA1c and proteic sulphur levels. Because the length of hair could provide in evolution a longer period than HbA1c (for every month hair raise with 1 cm), assays of proteic sulphur levels could give information about balance of diabetes for much more time than HbA1c. PMID- 15529633 TI - NIDDM: new once-daily intervention for type 2 diabetes mellitus: Diaprel MR. AB - Pharmacological treatment of hyperglycemia should address to both abnormalities in T2DM treatment, that is reduction of insulin resistance and restoration of normal insulin secretion. Gliclazide is a sulfonylurea compound oral hypoglycemic drug that has a unique feature of restoring the first-phase insulin secretion, which is lost in T2DM being one of the early features of disease. MATERIAL AND METHOD: Laboratoires Servier conducted in Romania a an open non randomized surveillance on the efficacy and safety of Diaprel MR in type 2 diabetic patients. 199 patients in 14 treatment centers were enrolled. Eligibility criteria were as it follows: men and women with diabetes, previously on diet alone and not treated with other OAD, over 35 years old with FPG (mg/dl) at enrollment between 126 and 180. The clinical trial lasted for 16 weeks. During this period the doctors examined the patients 6 times. First visits were at a 2 weeks interval and the last two visits at a 4 weeks interval. At each visit the doctor renewed the prescription for the subsequent period according to the following protocol: the starting dose was 30 mg Diaprel MR/day, if the FPG (mg/dl) was over 140 (at the next visit) the dosage was increased with 30 mg Diaprel MR/day, if the FPG (mg/dl) was under 140 the dosage remained the same as the previous dosage. The maximum dosage was 120 mg Diaprel MR/day. The following parameters were measured on first and last (seventh) visit: blood pressure (systolic and diastolic), heart rate (bpm), body mass index-BMI (kg/m2), fasting plasma glucose FPG (mg/dl and mmol/l), glycated hemoglobin HbA1c (%) and Hb-Hct (mg/dl-%), creatinine (mg/dl), SGPT (UI/I), cholesterol (mg/dl) and triglycerides (mg/dl). Blood pressure (systolic and diastolic), heart rate, BMI and FPG were measured from the second to sixth visit also. On each visit there was registered other data such as: associated illnesses, concomitant medication and adverse events. RESULTS: Primary end points. The average values of end points HbA1c (%) and PFG (mg/dl) registered a significant decrease during the 16 weeks of medication, from the enrollment moment (S0) to the last week (S16). The decrease was significant on the total sample of the main analysis group but also on subsamples of age, gender and BMI. HbA1c (%) average values decreased in the main analysis group (S16 compared to S0): with 22% on the total sample (from 7.7 to 6.0); p < 0.05. FPG (mg/dl) average values decreased in the main analysis group (S16 compared to S0): with 21% on the total sample (from 159 to 126); p < 0.05. Secondary end points. There were no significant changes registered in the average level of cholesterol and triglycerides, BMI, diastolic blood pressure, heart rate, creatinine, SGPT. A significant decline of the average systolic blood pressure was registered. CONCLUSION: Diaprel MR can be used safely in diabetic patients newly diagnosed, uncontrolled on diet or other oral antidiabetic drugs, overweight, safely in those with cardio-vascular disease, or in patients with creatinine clearance 50-80 ml/min. PMID- 15529635 TI - Toxic-induced hypoglycemia in clinical practice. AB - Toxic induced hypoglycemia is usually caused by the anti-diabetic treatment and excessive alcohol consume. Hypoglycemia in diabetics treated with insulin or anti diabetic oral agents is far the most studied form of hypoglycemia. Less information is available on toxic-induced hypoglycemia in non-diabetic subjects with acute exogenous poisoning. MATERIAL AND METHODS: We retrospectively studied adult non-diabetic patients admitted in Emergency Clinic Hospital of Iasi with hypoglycemia caused by an acute poisoning, over a period of 10 years. Then we performed a prospective study in those poisoning associated with hypoglycemic risk, to assess the prevalence of toxic-induced hypoglycemia. RESULTS: We identified 15,497 patients with acute poisoning in our retrospective study, 4,005 of whom presented poisoning associated with hypoglycemic risk (40% acute ethanol poisoning, 29% wild mushroom poisoning, 23% beta-blocker poisoning, 7% salicylate poisoning and 1% patients with anti-diabetic agents acute poisoning). The prospective study identified 1,034 patients with acute poisoning, 20.11% of whom had ethanol poisoning, 11.79% had beta-blocker poisoning, 5.89% had wild mushroom poisoning, 1.74% had salicylate poisoning and the rest of 60.47% had other acute poisoning, without hypoglycemic risk. Attempted suicide with anti-diabetic agents in non-diabetic subjects produced the most severe and prolonged form of hypoglycemia in acute poisoning. 23 patients in retrospective study and 6 patients in prospective study died, but in only one situation, the death was the direct consequence of hypoglycemia. CONCLUSIONS: In non-diabetic subjects with acute poisoning, prevalence of toxic-induced hypoglycemia depends on the poison itself, the mechanism of poisoning, also depends on the association between toxics and the severity of toxic- induced liver disease. A useful test to assess toxic-induced hypoglycemia is standard 6 hour oral glucose tolerance test (OGTT). Factors predicting a negative outcome in toxic-induced hypoglycemia are association of toxins, cardiac and hepatic complications, and age (>65 years). PMID- 15529636 TI - Alpha-lipoic acid in the treatment of autonomic diabetic neuropathy (controlled, randomized, open-label study). AB - AIM: to evaluate the effect of alpha-lipoic acid in autonomic diabetic neuropathy in a controlled, randomized, open-label study. MATERIAL AND METHODS: 46 patients with type 1 diabetes and different forms of autonomic neuropathy, of mean age 38.1 +/- 12.5 years and mean duration of diabetes 16.8 +/- 8.9 years were treated with alpha-lipoic acid for 10 days 600mg daily iv, thereafter one film tablet of 600mg daily for 50 days. 29 type 1 diabetic patients with autonomic diabetic neuropathy, of mean age 40.2 +/- 9.3 years and mean duration of diabetes 15.4 +/- 7.9 years served as a control group. We have followed-up patients' complaints, Ewing's tests, laboratory parameters of oxidative stress. RESULTS: There was a significant improvement after treatment in the score for severity of cardiovascular autonomic neuropathy--from 6.43 +/- 0.9 to 4.24 +/- 1.8 (p<0.001), while in the control group it worsened from 6.18 +/- 1.3 to 6.52 +/- 0.9 (p>0.1). We found improvement in the Valsalva manoeuvre after treatment - from 1.05 +/- 0.04 to 1.13 +/- 0.08 (p<0.001); in the deep-breathing test -from 3.4 +/- 2.8 to 10.4 +/- 5.7 (p<0.001); and in the lying-to-standing test--from 0.99 +/- 0.01 to 1.01 +/- 0.02 (p>0.1), while in the control group there was no improvement. There was a beneficial effect of treatment on the change of systolic blood pressure at the lying-to-standing test--from 22.7 +/- 11.5 to 9.8 +/- 7.9 (p<0.001), while in the control group the change was 20.5 +/- 11.1 mmHg and 19.7 +/- 12.9 mmHg (p>0.1), respectively. We found improvement in diabetic enteropathy in six patients; in the complaints of dizziness, instability upon standing in six patients; in neuropathic edema of the lower extremities in four patients and in erectile dysfunction in four patients after treatment, while in the control group no change was reported in the symptoms and signs of autonomic neuropathy by the end of the follow-up period. There were changes in the laboratory parameters of oxidative stress after therapy--total serum antioxidant capacity increased from 20.42 +/- 1.8 to 22.96 +/- 2.3 microgH2O2/ml/min (p<0.05), serum SOD activity - from 269.8 +/- 31.1 to 319.8 +/- 29.IU/l (p=0.02) and erythrocyte SOD--from 0.89 +/- 0.10 to 1.11 +/- 0.09 U/gHb (p=0.04). CONCLUSION: Our results demonstrate that alpha-lipoic acid (Thiogamma) appears to be an effective drug in the treatment of the different forms of autonomic diabetic neuropathy. PMID- 15529638 TI - Evidence-based research. PMID- 15529637 TI - Insulin pump therapy in pregnant women with type 1 diabetes mellitus. PMID- 15529639 TI - Congenital cholesteatoma. PMID- 15529640 TI - An inflammatory polyp above the torus tubarius as a cause of persistent serous otitis media. PMID- 15529641 TI - Airway foreign body. PMID- 15529642 TI - An unusual primary intratympanic meningioma. PMID- 15529643 TI - Chondrosarcoma of the larynx. PMID- 15529644 TI - Vestibular findings in a patient presenting with tinnitus. PMID- 15529645 TI - Percentage of reimbursement per CPT code. PMID- 15529646 TI - Nostril stenosis secondary to a laser burn injury: correction with a composite graft. PMID- 15529647 TI - Sequelae of undiagnosed cervical vertebral osteomyelitis in intravenous drug abusers. PMID- 15529648 TI - Modern acoustic reflectometry: accuracy in diagnosing otitis media with effusion. AB - We conducted a study of 80 patients to evaluate the accuracy of a commercially available acoustic reflectometer in identifying the presence or absence of otitis media with effusion (OME). This device assesses bilateral tympanic membrane mobility and, by inference, middle ear status. We found that it was most accurate in patients with normal and grossly fluid-filled ears. We recommend screening with this acoustic reflectometer to rule out OME in adult and pediatric patients. PMID- 15529649 TI - Ototopical ciprofloxacin in a glycerin vehicle for the treatment of acute external otitis. AB - We conducted a prospective, randomized, controlled, double-blind study of 33 patients to compare the efficacy and tolerability of a new glycerin formulation of ototopical 0.3% ciprofloxacin with that of a conventional aqueous formulation of ciprofloxacin for the treatment of acute external otitis. Outcomes measures were resolution of discharge, swelling, pain, and redness and the incidence of adverse side effects. Patients were examined on three occasions: on the day of enrollment (visit 1), 48 to 72 hours later (visit 2), and 7 days after enrollment (visit 3). At visit 2, the patients in the glycerin group showed a significantly greater resolution of discharge. We observed the same pattern with respect to swelling, pain, and redness, which resolved more quickly in the glycerin group, although not significantly so. All patients were cured by visit 3, and the two treatments were equally well tolerated. On the basis of our findings, we conclude that the glycerin formulation of ototopical 0.3% ciprofloxacin appears to be at least as effective as the aqueous form in the treatment of acute external otitis- and in the case of otorrhea, more so. PMID- 15529650 TI - Selective surgicel packing for the treatment of posterior epistaxis. AB - Rod lens endoscopes provide residents in otolaryngology a means of more accurately identifying the site of bleeding and, when possible, cauterizing the bleeding vessel. Identification of a posterior bleeding point is often difficult and sometimes impossible. Intranasal manipulation for electrocautery is painful, may require general anesthesia, and is associated with complications. We describe a pilot study designed to evaluate selectively packing the bleeding site with Surgicel (oxidized cellulose) to control the hemorrhage without packing the nasal cavity and to reduce patient morbidity and length of stay in the hospital. We describe the technique and present the results of treating 8 patients admitted with acute posterior epistaxis over a 10-month period in 1995-1996. PMID- 15529651 TI - Second branchial cleft anomaly presenting as a rudimentary pinna in the nasopharynx of a newborn. AB - We describe the case of a 4-day-old girl who presented with an epiglottic cyst that was later identified as a rudimentary pinna attached to the soft palate. PMID- 15529652 TI - Various presentations of fourth branchial pouch anomalies. AB - Embryologic anomalies of the fourth branchial pouch are rarely seen. They usually present as recurring episodes of deep neck infections and/or abscesses or acute suppurative thyroiditis. Failure to recognize these unusual cases may result in misdiagnosis, inadequate treatment, and subsequent recurrence. We report 3 cases of patients with fourth branchial anomalies. Diagnosis starts with a preoperative evaluation consisting of a barium swallow or sonogram followed by direct hypopharyngoscopy at the time of surgery. Treatment of acutely infected sinuses is best done with appropriate antibiotics and, if necessary, with incision and drainage. Surgical excision should be planned after the inflammation has completely resolved. PMID- 15529653 TI - Pleomorphic adenoma of the base of the tongue. AB - We describe a rare case of pleomorphic adenoma involving the base of the tongue and adjacent valleculae. We also discuss the incidence, diagnosis, and treatment of these cases. PMID- 15529654 TI - [Erosion in early school-age children]. AB - The aim of the present study was to determine the prevalence, the severity and the distribution of erosive lesions in children, living in a rural region in Switzerland. A total of 42 children, aged between five and nine years, were examined. The following parameters were assessed: erosion on the facial, occlusal and oral tooth sites; flow rate and buffering capacity of resting saliva; oral hygiene and nutrition. The occurrence of erosive lesions was as follows: All children had one or more erosive lesions degree 1 (erosion within enamel) on occlusal surfaces and 20 (47.6%) of them showed at least one lesion of degree 2 (dentinal erosion). Already six (14.3%) of the examined children had one or more erosive lesions on occlusal surfaces of permanent teeth. Facial and oral erosions were scarce and only deciduous teeth were involved: Facial erosive lesions were examined in four (9.5%) (degree 1) and two (4.8%) (degree 2) of the children and oral lesions in three (7.1%) (degree 1) and one (2.4%) of them (degree 2). No statistically significantly impact of the following parameters on the occurence, distribution and degree of erosive lesions was found: age, gender, flow rate of resting saliva, buffering capacity, orale hygiene and nutrition. The results show clearly that erosive lesions can be detected already on deciduous teeth and the young permanent dentition is also involved. Early detection of the lesions is important. Preventive measures have to be initiated to stop or at least to diminish the erosive process. PMID- 15529655 TI - [Bitewing examinations as a part of preventive dentistry--a review]. AB - For an optimal treatment of approximal caries lesions it is important to rely on a safe diagnosis. Bitewing examinations are an aid for detecting hidden lesions in the approximal surfaces. Other techniques for this purpose are fibre-optic transillumination (FOTI), digital imaging fibre-optic transillumination (DIFOTI), electric resistance measurement, laser fluorescence measurement and tooth separating. Since no direct view on approximal surfaces is feasible the detection of approximal lesions in the clinical examination is mostly insufficient. For a systematical controlling of the different grades of caries lesions, a time schedule for examinations is useful. For such a schedule, a couple of factors, such as caries risk of the patient or fluoridations, seem to be important. This article gives a survey of recommendations found in the literature to optimise the arrangement of bitewing examination in the therapeutical concept of caries monitoring and treatment planning. PMID- 15529656 TI - [Influence of magnification tools on the recognition of simulated preparation and filling errors]. AB - 37 mistakes or filling defects were mounted onto a phantom model. Three groups- each consisting of thirteen dentists--examined the jaws under clinical conditions using either no visual aid, magnifying glasses or a microscope. They were further asked if using magnifying tools had a positive effect on pains such as neck and back pain, headaches or sore eyes. The group using the microscope spent more time on examination and found significantly more defects than their colleagues using magnifying glasses. They also profited more from the ergonomical advantages. The main clinical use for microscopes is in endodontics. The group of dentists using magnifying glasses spent less time on examination but found more defects than their colleagues using no magnification tools at all. The positive effect on neck and back pain was less pronounced than in the group working with microscopes. Magnifying glasses are used in all kinds of clinical work. PMID- 15529657 TI - [Implant prosthesis rehabilitation after maxillofacial injuries--case report]. AB - The dental and oral rehabilitation of patients with severe trauma that involves the oral cavity is demanding and challenging. Recently, advanced surgical techniques and innovations in oral implantology led to more sophisticated treatment modalities that may better fulfill the patients needs in special situations. Young people rather often suffer from accidents--at school and during leisure activities--in a period of their life that has a great impact on their physical, mental and personal development. Thus, the wish for a quick functional and esthetical rehabilitation contrasts with the complex clinical situation and the compromised oral conditions. All persons involved in the rehabilitation process--including the patient--must seek for the best solution that takes into account treatment time, invasivity of procedures and patients morbidity. Further, it is also observed that insurances very much stress the reduction of cost and the simplicity of treatment. This will also influence the treatment planning and the sequence of the procedures as shown by the following case. PMID- 15529658 TI - [Long-term treatment of patients with heart defects. When is a new valve needed?]. PMID- 15529659 TI - ["Insurance should not any longer pay all"]. PMID- 15529660 TI - [Pregnancy discontinuance in an immunosuppressed patient. "A forbidden intervention can never be "indicated""]. PMID- 15529661 TI - [A tick sticks in the eyelid. How should it be removed?]. PMID- 15529662 TI - [Diagnosis and treatment of tumors in old age--few studies, no standards]. PMID- 15529663 TI - [Is oncological surgery contraindicated in the older patient?]. AB - Almost three-quarters of patients with cancer of the gastrointestinal tract are more than 65 years of age. In the operative treatment of esophageal pancreatic, hepatocellular and colorectal carcinomas, studies show comparable risks for patients under, and those over, 70 years of age. In terms of morbidity, mortality and 5-year mortality rates, attendant risks in the latter do not generally appear to be greater. An increase in the rate of complications, however, has been observed for palliative interventions in colorectal cancer patients. In particular cardiovascular risk factors play a major role in the over-70-year olds. PMID- 15529664 TI - [Radiotherapy in the elderly]. AB - Radiotherapy in curative or palliative intent is well tolerated and effective in the elderly patient, too. When establishing the indication for treatment, the extent of concomitant diseases, the general state of health of the patient, and his/her life expectancy must all be taken into account. When extensive volumes in the head/neck, chest or abdominal areas need to be irradiated, adequate uptake of fluid and calories must be ensured, and electrolytes, renal parameters and the blood count must be closely monitored. Supporting measures should be applied early on. PMID- 15529665 TI - [Medical treatment of cancer in the elderly]. AB - Paralleling increasing life expectancy, the number of cases of cancer in our population is also on the increase. The question as to whether chemotherapy for the treatment of a tumor in the elderly is appropriate or not, and if so what doses are required, depends in particular on the potentially increased toxicity of the drug in the old patient. The consequence is that alternative substances may have to be sought. When planning treatment, however, additional factors have to be taken into account, such as the possibility of differences in the physiology of the tumor in the elderly, the general state of health of the patient, the presence of concomitant diseases, possible interactions with other medications, and the patient's social environment. PMID- 15529666 TI - [Rendu-Osler-Weber syndrome--a complex systemic disease]. AB - Hereditary hemorrhagic teleangiectasia (HHT or Rendu-Osler-Weber Syndrome) is an inherited autosomal dominant disorder of the vascular connective tissue. The resulting vascular malformations can occur in virtually any organ. Nosebleeds can massively impact on the quality of life of those afflicted. However, visceral manifestations are likely to be more serious, and may be comparable with a "ticking time bomb". Most commonly affected are the vascular systems of the lungs, liver, brain and gastrointestinal tract. Screening is recommended--at least with regard to the lungs. Difficult constellations of this complex condition may be successfully managed by an interdisciplinary approach. PMID- 15529667 TI - [Headache in the elderly]. AB - Although the elderly generally suffer less often from headache, symptomatic headache and concomitant diseases are more frequent. For the diagnostic work-up of newly occurring headaches in old age, imaging examinations of the head (CT or MRI), together with laboratory investigations are necessary. The treatment of symptomatic headache is oriented to the underlying disease, and should be applied promptly, with the aim of preventing serious complications. When considering the management strategy, account must be taken of possible changes in pharmacokinetics, pharmacodynamics and the therapeutic response in the elderly. Numerous medications often cause headache or other side effects in patients of advanced age. Most antiheadache drugs have not been adequately been investigated in the group of over 65-year-olds. This means that drug treatment for headache is often applied in accordance with the principle: start low, go slow. PMID- 15529668 TI - [Bayern-kV is pleased with the new fee contract. But on line physicians get less money]. PMID- 15529669 TI - [Mistaken in buying a computer? So defend yourself correctly]. PMID- 15529677 TI - [Studies confirm: nightly micturition frequency lowered]. PMID- 15529676 TI - [Nightly polyuria. Adequate treatment improves efficiency and vitality during the day]. PMID- 15529678 TI - [Liver round foci: malignant or harmless?]. PMID- 15529679 TI - [A hairy condition]. PMID- 15529680 TI - [Voluminous medical record--no findings. Traveling pain from the psyche]. PMID- 15529681 TI - [Early summer meningoencephalitis. Only vaccination offers protection]. PMID- 15529682 TI - [AT1 blocker or calcium antagonist in treatment of hypertension. What protects the heart better?]. PMID- 15529684 TI - [Chronic vomiting and rapid weight loss. What compresses the small intestine?]. PMID- 15529683 TI - [Sequelae of the CARDS Study. LDL of 110 mg/dl still too high?]. PMID- 15529685 TI - [Ingrown toe nail. Little success with filing and surgery]. PMID- 15529686 TI - [Hypnotherapy. Highly effective techniques from Switzerland]. PMID- 15529687 TI - [Surgical correction of refractive errors. How risky is laser treatment of the eye? (interview by Verena Repp)]. PMID- 15529688 TI - [For forgetful or treatment noncompliant patients. Motivation tricks of colleagues (interviews by Dr. Judith Neumaier)]. PMID- 15529689 TI - [Bicycles for paraplegics--Hand-arm prostheses--artificial lenses. New developments in prosthetics use nature as a model]. PMID- 15529690 TI - [The electrical stimulation bicycle: a neuroprosthesis for the everyday use of paraplegic patients]. AB - Until recently, few patients with complete paraplegia could walk or stand with the help of functional electrical stimulation (FES) of the leg muscles regularly at home. In comparison, FES cycling with an adapted tricycle is easy to put into practice because the legs remain connected to the pedals and through the use of a tricycle or stationary bicycle, the balancing problems of the patient recedes into the background. In the first German feasibility studies for paraplegic cycling, eleven completely paraplegic patients have been tested so far. The goal is to make FES cycling a daily activity in the lives of as many patients as possible. PMID- 15529691 TI - [The artificial eye lens]. PMID- 15529692 TI - [Prosthetic possibilities after amputations in the upper extremities]. AB - Whereas passive prostheses are fitted onto the patient after the amputation of an arm, hand or finger and are mostly cosmetic in function, active prostheses have much more potential. They can transform the movements of other body regions to movement in the artificial limb. Belts or harnesses, for example, effect the direct transfer of the power from the muscle to the prosthesis. The range of movement possible depends upon the level of the amputation, the length of the residual limb, the age of the patient, his body build and fitness. Myoelectrically controlled prostheses possess their own drive and power source. They control movement through the electrical action potentials of the residual limb muscles, which are detected, amplified and transmitted with help of electrodes. PMID- 15529693 TI - [Significance of pedography in the diagnosis and treatment of the diabetic foot syndrome]. AB - Pedographic systems are now in use in many research centers and hospitals throughout the world. The platform system permits rapid and accurate static and dynamic measurement of the local pressure loading on the plantar surface of the foot, and also permits a functional assessment of the roll-over process in the bare foot. The in-shoe system employing an insole measures the pressure distribution within an orthopedic shoe and enables an accurate comparison of the loading situation before and after fitting. As an additional diagnostic tool, pedography improves preventive measures and the quality of the orthopedic treatment of the diabetic foot. PMID- 15529694 TI - [Recourse, medical errors, trapped fraud... Free legal counsel provides insight for physicians]. PMID- 15529695 TI - [Success of health care reform. Physicians save money for 57 KBV representatives]. PMID- 15529696 TI - [Transient ischemic attacks (TIA)]. PMID- 15529697 TI - [Diagnosis at a glance. Physician consultation not necessary?]. PMID- 15529698 TI - [Proctologic diseases: most etiologies are visible or palpable]. PMID- 15529699 TI - [Creatinine value alone is too unreliable. This formula reliably unmasks kidney damage]. PMID- 15529700 TI - [STIKO (established vaccination committee) recommends varicella vaccination for all children: are chickenpox really so risky? (interview by Dr. Thomas Meissner)]. PMID- 15529701 TI - [Quality control in oncology. Document any incidents!]. PMID- 15529702 TI - [Early and late complications of chemotherapy of solid tumors]. AB - Treatment with cytostatic agents is associated with both acute side effects such as nausea, emesis, blood count changes, and allergic reactions, and late complications such as cardiac, renal or gastrointestinal toxicity. In addition to differences in substance groups, intra-individual variations on the part of patients, and their general and nutritional status, are of particular importance for the severity of such side effects. For improved treatment monitoring, side effects should be recorded and documented in accordance with common toxicity criteria. In view of the complexity of oncological treatment, including the optimal supporting measures, such treatments should remain in the hands of oncological specialists. PMID- 15529703 TI - [Treatment of perivascular extravasation of cytostatic agents]. AB - In the intravenous administration of cytostatic drugs, top priority is given to preventive measures, and to immediate countermeasures in the event of an accidental paravenous injection. In contrast to earlier recommendations, the use of antidotes has been minimized. Only those with proven efficacy and no tissue toxic potential are to be used. The intradermal application of sodium bicarbonate and sodium thiosulfate, and the subcutaneous administration of glucocorticoids are no longer applicable. As a specific antidote in the case of anthracycline, topical dimethylsulfoxide (DMSO) is used, and for perivascular injection of vinca alkaloid, subcutaneous hyaluronidase is administered. PMID- 15529704 TI - [Acute and chronic side effects of local radiotherapy]. AB - Local radiotherapy of a tumor is frequently applied in combination with chemotherapy or surgical measures. Possibly occurring toxicity must therefore be considered within the overall context. The radiosensitivity of the various organs is dependent on the sensitivity of the respective stem or mature end cells, and shows considerable differences within the individual cell systems. Apart from the application of prophylactic procedures, some radiation sequelae can be ameliorated by initiating specific measures (e.g. cosmetic products, drug treatment). PMID- 15529705 TI - [Mycotic infection--allergic reaction--metastasis? Mysterious cutaneous rings]. PMID- 15529706 TI - [Yellow fever--a tropical danger]. AB - Yellow fever is an acute life-threatening viral infection. In the case of the urban type, the virus is transmitted from one human to another by the Aedes mosquito vector. In particular travelers to Africa from non-endemic areas run a high risk of clinical disease in case of infection. For prophylaxis, a reliable live virus vaccine is available. PMID- 15529707 TI - [Is endothelial dysfunction of practical relevance?]. AB - Endothelial dysfunction is a "systemic disease" and a predictor of preclinical atherosclerosis. A relatively simple-to-perform and reliable diagnostic method of evaluating endothelial function is the measurement of the forearm blood flow (FBF). Unremarkable vasoreactivity of the brachial artery on performing FBF measurement is a prognostically favorable sign. This applies equally to patients with cardiovascular risk factors such as hypertension, hyperlipidemia or diabetes mellitus, and to patients with angina pectoris. A number of investigations suggest that cardiovascular endpoints can be significantly diminished by reducing the risk factors. PMID- 15529708 TI - [Controlled release oxycodone--a new option in the treatment of severe and very severe pain]. PMID- 15529709 TI - [Family physicians are reimbursed 1,130 euro per patient. From this they must pay all colleagues--and the hospital]. PMID- 15529710 TI - [All offer quality management for physicians: doing it simply by yourself]. PMID- 15529711 TI - [Immunosuppression with sirolimus so donor kidneys survive longer]. PMID- 15529712 TI - [Donor organs still in short supply. Encountering death on the waiting list]. PMID- 15529713 TI - [GMG proposal for non-prescription medications. Expensive cost savings]. PMID- 15529714 TI - [Vascular symptoms are not confined to the legs. Peripheral arterial occlusive disease especially threatens the diabetic heart]. PMID- 15529715 TI - [Minor rate of side-effects makes it possible. Coxibs accrue new indications]. PMID- 15529716 TI - [Patient feels completely well. The PROCAM Score first reveals the high infarct risk]. PMID- 15529717 TI - [Rest, moisture and cortisone. First aid in pseudocroup]. PMID- 15529718 TI - [Stability also depends on other parameters. Bone density--not a good indicator for fracture prevention]. PMID- 15529719 TI - [Are there differences in bisphosphonates? (interview by Dr. Beate Schumacher)]. PMID- 15529720 TI - [Test for microalbuminuria. Life saving for hypertensive and diabetic patients]. PMID- 15529721 TI - [In moderate depression. St. John's wort maintains effectiveness]. PMID- 15529722 TI - [How does it work? Cardiac pacemaker in bradycardia]. PMID- 15529723 TI - [Red back, not delightful]. PMID- 15529724 TI - Economics and the education of nurse anesthetists: part 1. PMID- 15529725 TI - Expert testimony. PMID- 15529726 TI - Rave drugs: pharmacological considerations. PMID- 15529727 TI - Rural anesthesia: a pilot study. PMID- 15529728 TI - Diagnosis and treatment of negative pressure pulmonary edema in a pediatric patient: a case report. AB - This article includes a case study of a pediatric patient who presented for elective correction of a speech impediment via a frenuloplasty. The patient's history and anesthetic course will be discussed prior to the deveopment of the episode of negative pressure pulmonary edema (NPPE). The events surrounding the development and diagnosis of NPPE will be presented, and the treatment modalities will be discussed. PMID- 15529729 TI - Analgesia and effectiveness of levobupivacaine compared with ropivacaine in patients undergoing an axillary brachial plexus block. AB - A common anesthetic technique for the upper extremity is local brachial plexus anesthesia using levobupivacaine and ropivacaine. To our knowledge, no study has been performed measuring differences in analgesic efficacy and latency when these local anesthetics are used for brachial plexus anesthesia. We enrolled 54 adults, assessed as ASA class I or II, into this double-blind, prospective investigation to receive 40 mL of 0.5% ropivacaine or levobupivacaine with 1:200,000 epinephrine. Pain was assessed using a 0 to 10 verbal numeric rating scale (VNRS). Motor blockade was determined using a modified Bromage scale. Variables included analgesic duration, latency, and overall patient satisfaction. The ropivacaine group had significantly higher VNRS scores at the 8th (P= .001) and 10th (P = .003) postoperative hours. The duration of sensory analgesia was significantly longer in the levobupivacaine group (831 minutes) than in the ropivacaine group (642 minutes, P = .013). Return of motor activity was significantly faster in the ropivacaine group (778 minutes) than in the levobupivacaine group (1,047 minutes; P = .001). No other significant differences were noted between the groups. When considering levobupivacaine and ropivacaine for brachial plexus anesthesia, levobupivacaine should be considered when postoperative analgesia is a concern but not when an early return of motor activity is required. PMID- 15529730 TI - The use of the laryngeal mask airway with mechanical positive pressure ventilation. AB - The purpose of this article is to discuss the benefits, safety, and efficacy of the laryngeal mask airway (LMA) and identify the risks and misconceptions associated with LMAs when used with positive pressure ventilation (PPV). Despite the abundance of supporting evidence that LMAs may be used successfully in a variety of age groups and surgical procedures using PPV, many anesthesia providers are still reluctant to choose an LMA when PPV is needed. This reluctance emerges from the misconception that when using an LMA with mechanical ventilation, there is an increased incidence of gastric insufflation, failed ventilation, and pulmonary aspiration. When compared to other airway adjuncts, however, the LMA is a safe, effective means of delivering ventilation under anesthesia. PMID- 15529731 TI - Continuous subcutaneous insulin infusion during general anesthesia: a case report. AB - Care of the patient with diabetes mellitus presents numerous challenges to the anesthesia practitioner. There is no perfect way to care for these patients nor are any 2 patients with diabetes exactly alike. With the advent of subcutaneous insulin pumps, the anesthesia practitioner has another tool to assist him or her in giving high quality care. This case study describes the anesthesia care provided to a patient with type 1 diabetes who wore his continuous subcutaneous insulin infusion (CSII) pump during general anesthesia for surgical repair of a herniated lumbar disk. Importantly, the anesthesia plan involved a collaborative effort with the patient. Blood glucose levels were stable throughout the perioperative period. Little or no extra work was required of the CRNA. This case showed that the CSII could be used to minimize perioperative fluctuations in blood sugar. Postoperatively, the patient expressed a high degree of satisfaction with the anesthetic. PMID- 15529732 TI - The risks of blood transfusions and the shortage of supply leads to the quest for blood substitutes. AB - A number of factors have combined to drive the interest in developing blood substitutes. These include the time-dependent decrement in stored blood biochemistry, the general shortage of the blood supply, and public awareness of the risks associated with allogeneic transfusions. Current literature on different blood substitutes was reviewed. The aim of this article is to help the reader understand the necessity of blood substitutes and to briefly describe blood substitutes that are in clinical trials. The need for oxygen-carrying blood substitutes is the driving force in multiple clinical trials. More research is needed to develop alteratives to allogeneic blood transfusion that are free of complications. PMID- 15529733 TI - AANA Journal course: Update for nurse anesthetists--Part 4--preoperative cardiac evaluation. AB - This AANA Journal course discusses the American College of Cardiology (ACC) and American Heart Association (AHA) guideline on perioperative cardiovascular evaluation for noncardiac surgery. The intent of the ACC/AHA guideline is to assist clinicians in clinical decision making by describing a range of generally acceptable approaches for the diagnosis, management, and prevention of cardiac diseases. Optimizing the anesthetic management of the cardiac patient undergoing noncardiac surgery is becoming increasingly important: as the percentage of Americans older than 65 years continues to grow, so does the prevalence of cardiac disease in this population. Simply accepting a preoperative cardiology clearance for the cardiac patient undergoing noncardiac surgery provides little information that can be used for risk assessment and management of anesthesia. While national practice patterns vary significantly, there is an important need to standardize cost-effective preoperative cardiac evaluation. By using evidence based studies, the ACC/AHA guideline delineates methods to objectively categorize cardiovascular risk and use data from the cardiology consultation to refine anesthetic management. Use of the guideline can lead to more efficient evaluation of the noncardiac patient with cardiac disease, which can decrease morbidity, mortality, and cost. PMID- 15529734 TI - College students and HIV/AIDS: more insights on knowledge, testing, and sexual practices. AB - In this study, the authors confirm and expand previous findings (M. Anastasi, R. G. Sawyer, & P. J. Pinciaro, 1999; J. E. Lewis & R. M. Malow, 1997) concerning college students' knowledge and perceptions about HIV/AIDS. Although students are knowledgeable about HIV/AIDS, they have little personal concern about becoming infected and do not take appropriate safe sex precautions. Findings of the present study show that gender, ethnic background, and knowing someone infected by HIV/AIDS influence students' level of concern about infection. In addition, older students and those who know someone infected by HIV/AIDS are more likely to have been tested. Results are compared with national surveys of other age groups, and the authors suggest specific intervention strategies for college students. PMID- 15529735 TI - Work centrality and post-award work behavior of lottery winners. AB - Individuals who had won the lottery responded to a survey concerning whether they had continued to work after winning. They were also asked to indicate how important work was in their life using items and scales commonly used to measure work centrality. The authors predicted that whether lottery winners would continue to work would be related to their level of work centrality as well as to the amount of their winnings. Individuals who won large amounts in the lottery would be less likely to quit work if they had relatively greater degrees of work centrality. After controlling for a number of variables (i.e., age, gender, education, occupation, and job satisfaction), results indicated that work centrality and the amount won were significantly related to whether individuals continued to work and, as predicted, the interaction between the two was also significantly related to work continuance. PMID- 15529736 TI - Motivation when optimism declines: data on temporal dynamics. AB - To contribute to a description of motivation variables across time, the authors examined temporal changes in these variables as the time to pursue a test-taking goal approached. In three samples, expected performance, grade-level standards, and perceived adequacy of effort decreased as the test time approached, but other indices of motivation did not always decrease. Data indicated that (a) there is a strong relationship between expectancies and implicit goal setting, (b) students may sometimes change goal levels and definitions of success to maintain their desire for chosen goals despite declining expectations, (c) effects of event proximity on goal-achievement expectations may be based on overestimating the adequacy of future effort, and (d) the degree to which expectancy and value predict motivation appears to vary with exam proximity. Results revealed a temporal complexity in motivation variables and in the relationships among them. PMID- 15529737 TI - Development and psychometric properties of the perceptions of parenting inventory. AB - In the present article, the author discusses the development and validation of a new instrument, the Perceptions of Parenting Inventory (POPI), designed to measure the multidimensional construct of perceptions of parenting. The psychometric properties and construct validity of the POPI, a self-administered questionnaire, were examined with two samples: young childless university students (N = 282) and adults from the general community, the majority of whom were parents (N = 252). Within both samples, the POPI subscales evidenced sound factor structure and adequate internal reliabilities and construct validity as determined by its ability to distinguish between individuals with differing attitudes and intentions toward parenting. The POPI appears to be a reliable and valid tool for measuring the perceptions of parenting, regardless of whether these perceptions are based on expectations or experience. PMID- 15529738 TI - Self, identity, and emotional well-being among Turkish university students. AB - The aim of the present study was to identify indigenous or emic domains of self descriptions and to investigate identity orientations among urban middle-upper socioeconomic status Turkish university students. The relationships between emotional wellbeing, self-descriptions, and identity orientations, as well as the gender differences in these variables, were also explored. The 205 participants (mean age of 20.35) found independent and relational-interdependent aspects of the self and identity more descriptive than the collective or interdependent aspects. Significant gender differences were found between independent and collective-interdependent aspects of the self. Specifically, women reported more Personal Identity and more Social Influencability and less Traditionalism and Openness and Creativity-related self-definitions than did men. Finally, emotional well-being was positively associated with all types of identity orientations as well as with independent and relational-interdependent domains of self descriptions. The results are discussed with reference to the studies of self concept in the literature and the existing social change in Turkish society. PMID- 15529739 TI - High pressure nmr study of dihydrofolate reductase from a deep-sea bacterium Moritella profunda. AB - We have investigated the effect of pressure and temperature on the structural and thermodynamic stability of a protein dihydrofolate reductase from a deep-sea bacterium Moritella profunda in its folate-bound form in the pressure range between 3 and 375 MPa and the temperature range between -5 and 30 degrees C. The on-line cell variable pressure 1H NMR spectroscopy has been used to analyze the chemical shift and signal intensity in one-dimensional 1H NMR spectra. Thermodynamic analysis based on signal intensities from protons in the core part indicates that the thermodynamic stability of Moritella profunda DHFR is relatively low over the temperature range between -5 and 30 degrees C (deltaG0=15.8 +/- 4.1 kJ/mol at 15 degrees C), but is well adapted to the living environment of the bacterium (2 degrees C and 28 MPa), with the maximum stability around 5 degrees C (at 0.1 MPa) and a relatively small volume change upon unfolding (deltaV= 66 +/- 19 ml/mol). Despite the relatively low overall stability, the conformation in the core part of the folded protein remains intact up to approximately 200 MPa, showing marked stability of the core of this protein. PMID- 15529740 TI - Pressure-temperature-induced denaturation of yeast phosphoglycerate kinase, a double-domain protein. AB - Pressure-induced denaturation of yeast phosphoglycerate kinase was studied at various temperatures, as a model double-domain protein, using intrinsic fluorescence, 4th derivative absorbance, CD, and DSC. A thermodynamic transition intermediate was observed in the pressure-denaturation, as was reported for the cold denaturation. From the different response of Trp and Tyr residues, as monitored by fluorescence and 4th derivative absorbance changes, the C-terminal domain carrying all the Trp residues seemed to exert structural changes at relatively lower pressure. A further structural change involving both domains was observed at higher pressures. The two-step changes occurred almost simultaneously during heat denaturation. PMID- 15529741 TI - Exploration of pressure-induced dissociation of pyruvate oxidase. AB - The dissociation of pyruvate oxidase (PO) caused by pressure up to 220 MPa at various conditions was explored by measuring the intrinsic fluorescence spectra and polarization. At 5 degrees C and pH 7.6 the standard volume change (deltaV0) and free energy upon dissociation of the enzyme is -220 ml/mol and 29.83 kCal/mol, respectively. It was found that FAD was irreversibly removed during the pressure-dissociation of the enzyme. A much smaller standard volume change (-153 ml/mol) and lower free energy (24.92 kCal/mol) of apo-pyruvate oxidase (apo-PO) compared with the native enzyme indicated that FAD played very important role in stabilizing the enzyme and significantly influenced the standard volume change. The substrate pyruvic acid can significantly stabilize the enzyme against pressure in spite the standard volume for the enzyme in this case has a big increase relative to the native enzyme. The comparison of the intrinsic fluorescence of the native and the activated enzyme obtained by limited proteolysis indicated that the physical separation of alpha-peptide from the enzyme only occurred when the subunits were dissociated from each other under pressure. PMID- 15529742 TI - Effects of high pressure on the solubility and growth kinetics of monoclinic lysozyme crystals. AB - Average growth rates of the (0 1 0) and (0 1 0) faces (R<0 1 0>) of monoclinic lysozyme crystals were measured in situ under 0.1 and 100 MPa. From the dependence of the growth rates on the lysozyme concentration, we determined the solubility of the crystal as a function of temperature at 0.1 and 100 MPa. The solubility increased with an increase in pressure. From the comparison between the growth rates under 0.1 and 100 MPa at the same supersaturation level, we found that the growth rates of the monoclinic lysozyme crystals kinetically increase with an increase in pressure. Supersaturation dependencies of the growth rates under 0.1 and 100 MPa were well fitted with a two-dimensional (2D) nucleation growth model of a birth-and-spread type. The fitting results suggest that the increase in the growth rates with pressure can be explained by the decrease in the average ledge surface energy of 2D island, the average distance between the kinks on a step and the activation energies in the incorporation processes of solute molecules. PMID- 15529743 TI - High pressure fourier transform infrared (FT-IR) spectroscopic studies on inducible nitric oxide (NO) synthase active site: a comparison to cytochrome p450CAM. AB - High pressure Fourier transform infrared (FT-IR) spectroscopy is performed for the first time to analyse the active site of inducible nitric oxide synthase (iNOSox) using the carbon monoxide (CO) heme iron ligand stretch mode (nuCO) as spectroscopic probe. A membrane-driven sapphire anvil high-pressure cell is used. Three major conformational substates exist in substrate-free iNOSox which are characterized by nuCO at approximately 1936, 1945 and 1952 cm(-1). High pressure favors the 1936 cm(-1) substate with a volume difference to the 1945 substate of approximately -21 cm3/mol. The pressure induced cytochrome P420 formation with a reaction volume of approximately -80 cm3/mol is observed. Arginine binding produces a very low nuCO at approximately 1905 cm(-1) caused by the H-bond from the substrate to CO. nuCO for the substates in the substrate-free and arginine bound proteins shift linearly with pressure which is qualitatively similar to the observation on cytochrome P450cam. The slightly smaller positive slope of the shift in substrate-free iNOSox compared to substrate-free P450cam is interpreted as a slightly lesser compressible heme pocket. In contrast, the significant slower negative slope for arginine-bound iNOSox compared to camphor-bound P450cam results from the different kind of interactions to the CO ligand (electrostatic interaction in P450cam, H-bond in iNOSox). PMID- 15529744 TI - Cooperativity and high pressure: stabilization of the R conformation of the allosteric aspartate transcarbamylase under the influence of pressure. AB - The allosteric enzyme aspartate transcarbamylase (ATCase) from E. coli shows homotropic cooperative interactions between its six catalytic sites for the binding of the substrate aspartate. This cooperativity is explained by the transition of the enzyme from a conformation which has a low affinity for aspartate (T state) to a conformation with high affinity (R state). The crystallographic structures of these two conformations are known to a resolution of 2.5 A and 2.1 A, respectively, and they reveal an important difference in the quaternary structure of the protein. Enzyme kinetics under high pressure were used to study the transition between the two states. It appears that in the presence of a low concentration of aspartate, conditions under which the enzyme is essentially in the T state, pressure promotes the transition to the R state, the maximal effect being observed at 120 MPa. This transition is accompagnied by a significant deltaV. This observation is in accordance with the change in the protein surface exposed to the solvent, and with the increased number of water molecules bound to the protein. Since the partial specific volume of the enzyme does not change significantly during the T to R transition, the negative deltaV is only related to the change in hydration of the protein. This result emphasizes a significant role of the protein-solvent interactions in this important regulatory conformational change. PMID- 15529745 TI - Effects of hydrostatic pressure on horse liver alcohol dehydrogenase (HLADH): a new way of analyzing kinetic study. AB - Oxidation of ethanol by horse liver alcohol dehydrogenase (HLADH) is monitored under pressure (0.1 MPa - 225 MPa). The pressure-induced modifications of catalytic activity are followed by plotting reaction velocities as a function of substrates concentrations in the traditional double reciprocal form: then, pressure is treated as an activator (p < 100 MPa) or an inhibitor (p<225 MPa). Surprising typical patterns of Lineweaver-Burk curves are observed and interpreted. These results suggest that this approach could be a powerful tool to study enzyme's structure-activity relationship. PMID- 15529746 TI - Pressure activation of the chaperone function of small heat shock proteins. AB - Small heat shock proteins play an important role in the stress response of cells and in several other cellular functions. They possess chaperone-like activity; i.e. they can bind and protect damaged proteins from aggregation and maintain them in a folding-competent state. Two members of this family were investigated in this work: bovine alpha-crystallin and heat shock protein (HSP)16.5 from the thermophilic archaebacteria Methanococcus jannaschii. We reported earlier the enhancement of chaperone potency of alpha-crystallin by high pressure. We now report the completion of the work with results on HSP16.5. The chaperone potency of both proteins can be enhanced significantly by applying high pressure. Evidence by light scattering, Fourier transform infrared (FT-IR) and tryptophan fluorescence experiments show that while the secondary and tertiary structure of these proteins are not influenced by high pressure, their quatemary structure becomes affected: H bonds between subunits are weakened or broken, tryptophan environments become more polar, oligomers dissociate to some extent. We conclude that the oligomeric structure of both proteins is loosened, resulting in stronger dynamics and in more accessible hydrophobic surfaces. These properties lead to increased chaperone potency. PMID- 15529747 TI - Trimethylamine oxide, betaine and other osmolytes in deep-sea animals: depth trends and effects on enzymes under hydrostatic pressure. AB - Most shallow teleosts have low organic osmolyte contents, e.g. 70 mmol/kg or less of trimethylamine oxide (TMAO). Our previous work showed that TMAO contents increase with depth in muscles of several Pacific families of teleost fishes, to about 180 mmol/kg wet wt at 2.9 km depth in grenadiers. We now report that abyssal grenadiers (Coryphaenoides armatus, Macrouridae) from the Atlantic at 4.8 km depth contain 261 mmol/kg wet wt in muscle tissue. This precisely fits a linear trend extrapolated from the earlier data. We also found that anemones show a trend of increasing contents of methylamines (TMAO, betaine) and scyllo inositol with increasing depth. Previously we found that TMAO counteracts the inhibitory effects of hydrostatic pressure on a variety of proteins. We now report that TMAO and, to a lesser extent, betaine, are generally better stabilizers than other common osmolytes (myo-inositol, taurine and glycine), in terms of counteracting the effects of pressure on NADH Km of grenadier lactate dehydrogenase and ADP Km of anemone and rabbit pyruvate kinase. PMID- 15529749 TI - Characterization of a complex formed between human plasminogen and recombinant sheep prion: pressure and thermal sensitivity of complex formation. AB - Scrapie is thought to be caused by one or more conformations of a proteinacious particle called a prion. The infectious form(s) is referred to as the scrapie form of the prion protein (PrPsc) whereas a benign form, the cellular conformer, is referred to as PrPC. The cellular conformation of the sheep prion protein formed a 1:1 complex with human plasminogen. The complex precipitated at 0 degrees C (Ksp = 17* 10(-12) M2). This precipitation reaction was sensitive to both temperature and pressure. When subjected to hydrostatic pressure the precipitate dissolved. At 25 degrees C the complex was soluble with a dissociation constant of about 10(-7) M as determined by isothermal titration calorimetry. Absorption, fluorescence and circular dichroism spectroscopy showed that neither protein, in the complex, underwent a detectable structural change so long as proteolytic inhibitors were present. In the absence of proteolytic inhibitors, plasminogen slowly cleaved the prion protein. PMID- 15529748 TI - High pressure, an alternative approach to understand protein misfolding diseases. AB - Protein folding is essential for the flow of genetic information to biological activity. A failure in this process can result in disease, by causing cell damage and sometimes death. The misfolding of proteins often induces their aggregation, initiating the fibril formation seen in a range of human and animal diseases. Because misfolding and aggregation are of fundamental importance in vivo, there is currently great interest in understanding their mechanisms. To gain insight into the folding and unfolding processes of proteins, for nearly a century, an original biophysical approach has been successfully used: the application of high hydrostatic pressure combined with various spectroscopic and kinetic techniques. Because high pressure provides new insight into protein structure and folding which cannot be obtained by other techniques, the conformations of pressure induced unfolding intermediates and species involved in the initial states of aggregation of proteins associated with specific diseases are currently being investigated. Our contention is that by exploring folding kinetics, misfolding pathways and stability under pressure, it will be possible to understand the mechanisms of amyloidogenesis, with the ultimate goal to design therapeutic strategies to prevent progression of the disease. PMID- 15529750 TI - Temperature-pressure configurational landscape of lipid bilayers and proteins. AB - High hydrostatic pressure has been used as a physical parameter for studying the stability and energetics of biomolecular systems, such as lipid bilayers and proteins, but also because high pressure is an important feature of certain natural membrane environments. By using a variety of spectroscopic and scattering techniques, the temperature and pressure dependent structure and phase behaviour of various lipid systems and proteins have been studied and are discussed. A thermodynamic approach is presented for studying the stability of proteins as a function of both temperature and pressure. Moreover, the effect of various chaotropic and kosmotropic cosolvents on the temperature- and pressure-dependent structure and stability of proteins is discussed. The results demonstrate that combined temperature-pressure-cosolvent dependent studies can help delineate the free energy landscape of proteins and hence help elucidate which features and thermodynamic parameters are essential in determining the stability of the native conformational state of proteins. We also introduce pressure as a kinetic variable. Applying the pressure-jump relaxation technique in combination with time-resolved synchrotron X-ray diffraction and spectroscopic techniques, the kinetics of un/refolding of lipid mesophases and proteins has been studied. Finally, recent advances in using pressure for studying misfolding and aggregation of proteins will be elucidated. PMID- 15529751 TI - Virus stability and protein-nucleic acid interaction as studied by high-pressure effects on nodaviruses. AB - In this work, we evaluate the stability, dynamics and protein-nucleic acid interaction in Flock House virus (FHV). FHV is an RNA insect virus, non enveloped, member of the family Nodaviridae. It is composed of a bipartite single stranded RNA genome packaged in an icosahedral capsid of 180 copies of an identical protein (alpha protein). A fundamental property of many animal viruses is the post-assembly maturation required for infectivity. FHV is constructed as a provirion, which matures to an infectious virion by cleavage of alpha protein into beta and gamma subunits. We used high pressure, temperature and chemical denaturing agents to promote perturbation of the viral capsid. These effects were monitored by spectroscopy measurements (fluorescence, light scattering and CD) and size-exclusion chromatography. The data showed that FHV was stable to pressures up to 310 MPa at room temperature. The fluorescence emission and light scattering values showed small changes that were reversible after decompression. When we combined pressure and sub-denaturing urea concentrations (1 M), the changes were more drastic, suggesting dissociation of the capsid. However, these changes were reversible after pressure release. The complete dissociation of FHV could be observed only under high urea concentrations (10 M). There were no significant changes in emission spectra up to 5 M urea. FHV also was stable when we used temperature treatments (high and low). We also compared the effects of urea and pressure on FHV wild type and cleavage-defective mutant VLPs (virus-like particles). The VLPs and authentic particles are distinguishable by protein-RNA interactions, since VLPs pack cellular RNA and native particles contain viral RNA. Our results demonstrated that native particles are more stable than VLPs to physical and chemical treatments. Our data point to the specificity of the interaction between the capsid protein and the viral RNA. This specificity is crucial to the stability of the particle, which makes this interaction an excellent target for drug development. PMID- 15529752 TI - Psychropiezophilic microorganisms. AB - Bacteria living in the deep-sea have several unusual features that allow them to thrive in their extreme environment. Most isolated strains are not only piezophilic but also psychrophilic. In this short review, we summarize the current knowledge about psychrophiezophilic microorganisms with regard to their taxonomy and cellular fatty acids composition. Their biotechnological potential is also described. PMID- 15529753 TI - Piezophysiology of yeast: occurrence and significance. AB - Hydrostatic pressure is a thermodynamic parameter that has recently received further consideration in various experimental fields. Although the physicochemical basis of pressure effects is well established, the effects of high pressure on in vivo biological processes have not been systematically investigated due to its presumed complexity. The word "piezophysiology" was proposed to describe the unique cellular responses to high pressure in living cells. The yeast Saccharomyces cerevisiae is one of the best-characterized organisms in many fields in bioscience. Here I review the accumulated literature on cellular responses to increasing hydrostatic pressure in yeast, focusing on survival, growth, gene expression and metabolism. PMID- 15529754 TI - Pressure response in the yeast Saccharomyces cerevisiae: from cellular to molecular approaches. AB - Yeasts are unicellular organisms that are exposed to a highly variable environment, concerning the availability of nutrients, temperature, pH, radiation, access to oxygen and, specially, water activity. Evolution has selected yeasts to tolerate, to a certain extent, these environmental stresses. High hydrostatic pressure (HHP) exerts a broad effect upon yeast cells, interfering with the cell membranes, cellular architecture and in processes ofpolymerisation and denaturation of proteins. Gene expression patterns in response to HHP revealed a stress response profile. The majority of the upregulated genes were involved in stress defence and carbohydrate metabolism while most of the repressed ones were in cell cycle progression and protein synthesis categories. In addition, in the present work it was seen that mild pressure induced cell cycle arrest and protection against severe stresses, such as high temperature, high pressure and ultra cold shock. Nevertheless, this protection was only significant if the cells were incubated at atmospheric pressure after the HHP treatment. Expression of genes that were upregulated by HHP and are related to resistance to this stresses were also analyzed, and, for the majority of them, higher induction was attained after 15 min post pressurization. Taken together, the results imply an interconnection among stresses. PMID- 15529755 TI - Hydrostatic pressure induced death of mammalian cells engages pathways related to apoptosis or necrosis. AB - We investigated the response to high hydrostatic pressure (HHP) of mammalian cells, since HHP is proposed to be suitable to inactivate mammalian cells in biopharmaceutics and patient's material. We observed that cells were not restricted in their viability by pressures up to 100 MPa. Mammalian cells die when treated with pressures of 200 MPa or more. But the effects of 200, 300 or 400 MPa do not follow the same pattem. At 200 MPa, cells die in a way that is related to apoptosis. Some apoptotic characteristics like phosphatidylserine (PS) exposure and morphological alterations appear very fast. Other features like a higher exposure of intracellular NPn ligands and pronounced degradation of DNA and lectin ligands are unique features of HHP induced apoptosis. Cells treated with 300 and 400 MPa die immediately following a unique necrotic pathway, since treated cells harbour high DNA and glycoprotein degrading activities. PMID- 15529756 TI - High hydrostatic pressure inactivated human tumour cells preserve their immunogenicity. AB - High hydrostatic pressure (HHP) is an established method to inactivate biomolecules and microoganisms. It is routinely used for the sterilization of foodstuff. Recently, new applications as inactivation of microorganisms and tumour cells for bone transplants or for cancer vaccines have emerged. Characterization of the HHP-induced cellular responses are a prerequisite for its clinical use. To this end, we investigated the fate of human cells after HHP by cytofluorometry. We observed that the induction by HHP of cell death is time- and pressure-dependent. Surprisingly, an HHP-treatment of 100 MPa did not reduce viability at any time point. Pressures from 150 to 250 MPa-induced programmed cell death in most cells. However, survivors were observed in long term culture experiments under these conditions. Pressures above 300 MPa immediately induced cell death by necrosis and completely inactivated the cells. In contrast to inactivation by other necrosis inducing treatments like heat, freeze/thaw, or chemical agents, HHP avoids generation of Maillard products and disintegration and lysis of the cells. Instead HHP generates a gelatinised mixture of antigens captured in a distinct and robust particle and maintains their humoral immunogenicity. The high viscosity of the internal matrix of a pressurised cell is reflected by the slow penetration of the low molecular compound propidium iodide and limits the bleeding of antigen before uptake by antigen presenting cells. Taken together, HHP is an alternative method for the inactivation of mammalian cells in clinical settings. PMID- 15529757 TI - High pressure microscopy--a powerful tool for monitoring cells and macromolecules under high hydrostatic pressure. AB - A high pressure chamber, which withstands a pressure up to 300 MPa has been developed. The so-called HPDS (Hartmann, Pfeifer, Dornheim, Sommer) High Pressure Cell in combination with an inverted microscope and an analysis system allows brilliant microscopic colour pictures with an optical resolution better than 0.56 microm. The pressure chamber allows the in situ observation of dynamic changes of microscopic structures in bright field, phase contrast and fluorescence microscopy. This publication should demonstrate the capabilities of the system using results of experiments with two types of Spirogyra algae. The pictures have shown significant variations of the chloroplasma and the cell wall membrane at pressures of up to 120 MPa. The new system provides a simple way to perform microscopic analyses at pressures of up to 300 MPa. PMID- 15529759 TI - Pressure and temperature interactions on cellular respiration: a review. AB - Thermodynamic equations show that pressure and temperature can, theoretically, act in synergy or in opposite directions depending on their respective variations. Hence, they interact to establish rates of biological processes (pressure/temperature interactions, PTI). For such studies, it is interesting to use aquatic ectotherms, in particular fish, because it is easy to submit them to temperature and/or pressure changes. This review focuses on the effects of temperature and pressure changes on the energy metabolism of fish, mitochondrial oxygen consumption and functioning, showing that the observed effects do not always match the predictions made by equations or models. Unpublished results concerning the mitochondrial function of eels acclimatised at two temperatures and two pressures show that the mitochondrial targets of pressure and temperature are probably not the same. The possible mechanisms and consequences of PTI are discussed. PMID- 15529758 TI - High hydrostatic pressure induces ERK and PI3 kinase phosphorylation in human HCS 2/8 chondrosarcoma cells. AB - High continuous hydrostatic pressure has been shown to affect many cellular functions within the pressurised cells, for instance, accumulation of heat shock protein 70 occurs during pressurisation. Various signal transduction pathways are likely to mediate these changes, however, at the present time our knowledge of the pathways involved is rather limited. The aim of this study was to investigate whether some of the well known transduction pathways are activated by the exposure of human chondrosarcoma cells to 15-30 MPa hydrostatic pressure. The results showed an increased presence of the active, phosphorylated forms of extracellular signal-related kinase (ERK) and phosphoinositide 3-kinase (PI3K) in cells exposed to 15 and 30 MPa continuous hydrostatic pressure, while 0.5 Hz cyclic loading had weaker effects. Inhibition of ERK-pathway with UO126 did not prevent the accumulation of heat shock protein 70. No activation of c-Jun N terminal protein kinase (JNK) or p38 could be noticed in pressurised cells. In conclusion, we could identify at least two different signal transduction pathways that are activated under high continuous hydrostatic pressure. Accumulation of heat shock protein 70 was independent of ERK-activation. PMID- 15529760 TI - CIOS feel the heat. Former "IT people" arriving at the executive table may find themselves on the hot seat of intensified demands. PMID- 15529761 TI - Shopping for potential. The search for practice management software should begin with EMR evaluation. PMID- 15529762 TI - Long-range forecasting. Predictive modeling's power to provide an accurate outlook is strong and growing. PMID- 15529763 TI - A vision of data sharing. When a network architect joins healthcare collaborators, a community benefits. PMID- 15529764 TI - Do you have an appointment? Automated systems ease the load for patient schedulers, and that's just the beginning. PMID- 15529765 TI - Contingency planning. HIPAA's security rule lists five implementation specifications. PMID- 15529766 TI - Guides and prompts for safety. Just out of the starting gate, decision support is expected to advance steadily. PMID- 15529767 TI - Server virtualization. There's a way around supporting multiple servers and operating systems. PMID- 15529768 TI - Bar code labeling. Healthcare requires disciplined, verifiable procedures and process management. PMID- 15529769 TI - [Neuroemergency: harmonization with neurosurgery and emergency medicine]. PMID- 15529770 TI - [Neuroanatomical study of the internal structures of the brain based on brain fiber dissection]. AB - From the beginning of the 17th Century, anatomists started to elucidate numerous tracts and fasciculi of the brain using fiber dissection techniques. Due to the complexities associated with tissue preparation and the time-consuming nature of traditional techniques the popularity of this method has gradually declined with the advent of modern neuroimaging techniques such as MRI. Many novel approaches to deep brain lesions have recently been devised following the development of skull base surgery. In comparison, the study of functional neuroanatomy has been relatively neglected. Understanding how neuronal fibers in the brain interconnect and communicate neuronal functions is critical when performing surgery for malignant glioma and epilepsy, as brain parenchyma has to be resected and neuronal fibers are interrupted. In the preoperative planning process we have applied anatomical studies using brain fiber dissection techniques. Brain fiber dissection offers the advantage of exposing association, commissural and projection fibers of cerebral white matter in three dimensions. Brain fiber dissection appears useful for assisting neurosurgeons to acquire neurosurgical skills and become familiar with neuroanatomical features. The techniques and significance of brain fiber dissection are described herein. PMID- 15529771 TI - [Unusual MR appearance of intracranial dissemination from cervical glioblastoma]. AB - We reported a rare case of cervical glioblastoma with intracranial dissemination at an early stage of clinical course and reviewed the literature. An 8-year-old girl presented with failure of vision 3 months prior to admission to our hospital. Neurological examination on admission disclosed no definitive abnormalities except for bilateral visual disturbance and optic atrophy. Cranial MR images revealed a homogeneously enhancing tumor in the left sylvian fissure. Multiple spotty T2-hyperintensity lesions without contrast enhancement were also disclosed in bilateral cerebellum. Spinal MR images showed an enhancing tumor at C7 and tiny enhancing lesions on the surface of T11. The patient underwent an uneventful excision of the exophytic tumor at C7. The histological diagnosis was glioblastoma. The enhancing tumor in the left sylvian fissure treated by Linac stereotactic radiotherapy with a marginal dose of 38.4Gy in 12 fractions has diminished, whereas the residual tumor at C7 remained unchanged after radiation of 44Gy. In contrast, the multiple spotty lesions without contrast enhancement dispersedly spread in the cerebellum and infiltrated into brain stem despite 4 courses of chemotherapy using ifosfamide, cisplatin, and etoposide. Stereotactic biopsy of the multiple spotty lesions in the cerebellum was performed. Histological examination revealed anaplastic astrocytoma. The patient died 2 weeks after the biopsy despite additional chemotherapy and focal irradiation to the cerebellum. Early detection and selection of optimal therapeutic strategies are important in management of spinal glioblastoma with subarachnoid dissemination, since neuroradiological findings and therapeutic sensitivity are varied according to differentiation of disseminated tumors. PMID- 15529772 TI - [Postoperative transient hemiplegia after resection of the medial frontal tumor involving the supplementary motor area: report of two cases]. AB - The supplementary motor cortex (SMA) is located anterior to the primary motor cortex, and is considered to play an important role in planning, initiating and maintaining sequential motor actions. Disturbance of this area sometimes causes severe contralateral hemiplegia. If the disturbance doesn't affect the primary motor cortex, motor function will recover in relatively early postoperative time. We encountered two cases in which the patients developed postoperative severe hemiplegia after resection of a medial frontal lobe glioma, although there was no apparent change shown in the monitoring of intraoperative motor evoked potential (MEP). Both cases recovered from hemiplegia in the early postoperative period. In our cases, the disturbances of SMA were considered to be the causes of the development of transient hemiplegia. Intraoperative MEP monitoring is useful to distinguish the damage of the primary motor area from that of the SMA. PMID- 15529773 TI - [Intracavernous iatrogenic aneurysm causing subarachnoid hemorrhage after removal of intracranial tumor, treated by coil embolizatioin]. AB - We describe a rare case of subarachnoid hemorrhage due to a ruputured iatrogenic traumatic aneurysm in the cavernous carotid artery, caused by injury during surgery for skull base meningioma that was performed 2 years ago. A 64-year-old woman underwent craniotomy for resection of meningioma of the right sphenoid ridge. During surgery, venous bleeding from the cavernous sinus was easily controlled by packing. Tumor infiltration into the artery had not occurred, and total resection was successfully performed. Two years later, the patient was admitted to our hospital for subarachnoid hemorrhage, without clinical signs of carotid cavernous fistula. Angiography displayed an aneurysm in the cavernous portion of the right carotid artery, which had not been detected on a previous angiogram. The aneurysm was successfully embolized with a GDC via an endovascular approach. Three months later, the residual aneurysm became enlarged and aneurysmal embolization was performed for a second time. Follow-up angiography was performed 7 months after initial embolization, and revealed complete packing. PMID- 15529774 TI - [A case of pleomorphic TSH-producing pituitary adenoma with calcification]. AB - We reported a rare case of pleomorphic TSH-producing pituitary adenoma with calcification and reviewed the literature. A 25-year-old female was admitted to our hospital with a complaint of anterior neck swelling. An endocrinological examination demonstrated elevated serum levels of free triiodothyronine (FT3: 5.6 pg/ml), free thyroxin (FT4: 2.2 ng/dl), TSH (5.85 microIU/ml), and TSH a-subunit (5.9 ng/ml), whereas a physical examination revealed no goiter. CT scan showed a suprasellar isodensity mass with dense calcification. Contrast-enhanced T1 weighted images revealed a less enhancing tumor extending from the left upper portion of pituitary fossa into suprasellar cistern. The patient underwent gross total removal of the tumor via the right pterional approach. Microscopically, medium-sized to enlarged tumor cells with marked pleomorphism and prominent calcification were observed. The tumor cells displayed positive reaction for TSH (beta-subunit). The MIB-1 index averaged 2.9%. The histological diagnosis was a pleomorphic TSH-producing pituitary adenoma. Postoperatively, the serum levels of FT3, FT4, TSH, and TSH alpha-subunit decreased to normal range. Follow-up MR images showed no evidence of recurrent tumor 3 years after the resection. All of six patients with densely calcified TSH-producing pituitary adenoma, previously reported in the literature, remained well without tumor recurrence. We suggest that this type of TSH-producing pituitary adenoma may be associated with favorable prognosis despite histologically pleomorphic appearance. PMID- 15529775 TI - [A case of superficial siderosis following subtotal removal of pituitary adenoma]. AB - Superficial siderosis of the central nervous system (SS) is a rare clinical syndrome due to repeated intracranial hemorrhage. We report a case of SS occurring 19 years after subtotal removal of a non-functioning pituitary adenoma. The patient was a 37-year-old female, exhibiting progressive bilateral sensory neural hearing loss and cerebellar ataxia. T2 weighted magnetic resonance imaging demonstrated a rim of low intensity signal on the cerebellum, brain stem, and sylvian fissure. Immediate diagnosis based on characteristic symptoms and MRI findings is important for the prevention of irreversible progression of SS. PMID- 15529776 TI - [Cell therapy for Parkinson's disease: a guide to stereotactic brain operations]. PMID- 15529777 TI - [Clinico-pathology of skull tumor: aneurysmal bone cyst, fibrous dysplasia, eosinophilic granuloma]. PMID- 15529778 TI - [Natural history of pancreatic carcinoma]. PMID- 15529779 TI - [Gene therapy for digestive cancer]. PMID- 15529780 TI - [The current status of virotherapy for gastrointestinal cancer]. PMID- 15529781 TI - [A case of primary malignant melanoma of the esophagus and analysis of 193 patients in Japan]. PMID- 15529782 TI - [A case of Zollinger-Ellison syndrome due to duodenal gastrinoma with incurable diarrhea which was led to diagnosis by endoscopic examination]. PMID- 15529783 TI - [A case of acute occlusion of the superior mesenteric artery after acute cholecystitis and liver infarction]. PMID- 15529785 TI - [A case of telangiectatic focal nodular hyperplasia with remarkable hemorrhage and necrosis]. PMID- 15529784 TI - [A case of AFP-producing cholangiocarcinoma with difficulty in differential diagnosis from hepatocellular carcinoma]. PMID- 15529786 TI - [Biliary cystadenocarcinoma from biliary cystadenoma]. PMID- 15529787 TI - [Concept and practice of Sugita's microneurosurgical system]. PMID- 15529788 TI - [Lumbosacral lipomas]. PMID- 15529790 TI - [A surgical case of subdural hematoma with hemophilia A]. AB - A 54-year-old man presented with unconsciousness. Computed tomography revealed acute subdural hematoma. Emergency evacuation of hematoma was performed showing any excessive tendency to bleed or difficulty to stop bleeding during the operation. However transfusion of fresh frozen plasma was needed to stop continuous bleeding from the surgical wound after the operation. The patient underwent craniotomy again 18 days after the operation because he suffered hemorrhagic infarction and recurrence of acute subdural hematoma. After the second operation, a coagulability examination revealed that his activated partial thromboplastin test was prolonged (74.5 seconds) and his plasma factor VIII level was 20% of normal, so he was diagnosed as having mild hemophilia A. Cranioplasty was accomplished with replacement therapy, and he was discharged with mild recent memory disturbance and homonymous hemianopsia. PMID- 15529789 TI - [Evaluation of brain tumors by simultaneous dual isotope SPECT with 201Tl chloride and 99mTc-MIBI]. AB - Single photon emission computed tomography (SPECT) is useful for detecting brain tumors. In this study, we evaluated the utility of simultaneous dual SPECT with 201Tl-Chloride (Tl) and 99mTc-MIBI (MIBI) for diagnosis of brain tumors. We evaluated 20 cases, including 2 glioblastomas, 7 anaplastic astrocytomas, 2 oligodendrogliomas, 2 anaplastic ependymomas, 2 medulloblastomas, 2 meningiomas, 1 malignant meningioma, 1 pituitary adenoma, and 1 craniopharyngioma. We analyzed the uptake ratio (T/N ratio) of tracers in both Tl and MIBI at max counts/pixels ratio in the region of interest. The T/N ratios in early and delayed images were described as early ratios (ER) and delay ratios (DR), respectively. The retention index (RI) was calculated as the DR/ER ratio. Significant correlations were found between ER and DR for both Tl (DR = 0.797 * ER + 0.359, r = 0.871), and MIBI (DR = 0.961 * ER - 0.191, r = 0.784). Next, we analyzed the correlations between Tl and MIBI SPECT, for ER, DR, and RI. ER values for the two were strongly correlated (r = 0.791), DR values were weakly correlated (r = 0.556), and RI exhibited no correlation between them (r = 0.328). There were no correlations between tumor volume and T/N ratio for the two (ER-Tl; r = 0.0095, DR-TI; r = 0.0050, ER-MIBI; r = 0.036, DR-MIBI; r = 0.254). Lastly no correlation was found between RI-Tl and RI-MIBI (r = 0.328). We discuss the difference in the mechanism of accumulation of two tracers and the significance of simultaneous dual SPECT using them for the differential diagnosis of pituitary tumors, regrowth of oligodendrogliomas, and multidrug resistance of chemotherapy. Dual SPECT with Tl and MIBI appears to be useful for the diagnosis of brain tumor. PMID- 15529791 TI - [Multiple cerebral aneurysms and a contralateral occluded internal carotid artery associated with persistent primitive trigeminal artery]. AB - The frequency of angiographic demonstration of persistent primitive trigeminal artery (PTA) is 0.06 to 0.6%. It is well recognized that approximately 13.8 to 27.8% of patients with PTA also have intracranial aneurysms. However their association with the contralateral occluded internal carotid artery (IC) has not been reported. We reported such an extremely rare case of PTA associated with multiple cerebral aneurysms and a contralateral occluded IC. A 61-year-old female suffered from sudden onset of severe headache with nausea and vomiting. When she was admitted to our hospital, she complained only of headache with nausea. Computed tomography demonstrated a diffuse subarachnoid hemorrhage. Angiography showed a ruptured anterior communicating artery (Acom). Aneurysm and a left IC-PC large aneurysm with a broad neck. The angiogram also demonstrated a left PTA originating from the cavernous portion of the left internal carotid artery and ending at the midportion of the basilar artery and the contralateral occluded IC in the cervical portion. An operation was performed at day 3 using the left pterional approach, and the two aneurysms were successfuly clipped using the suction-decompression method. The patient was discharged with no neurological deficits. PMID- 15529792 TI - [Case report of Rosai-Dorfman disease mimicking pachymeningitis]. AB - Rosai-Dorfman disease (RDD) is a rare idiopathic histioproliferative disease affecting the lymph nodes. Extranodal sites may be involved and occasionally represent the initial or sole manifestation of the disease. Central nervous system manifestations especially are exceedingly rare. We reported a case of isolated, intracranial, dural-based RDD. A 69-year-old male, admitted with a second generalized convulsion, was examined by high-resolution MR images that revealed a thickened, stratified dura in the left frontal region with associated cortical edema. The patient underwent craniotomy with subtotal resection of the lesion. Microscopically, with the cytoplasmic staining against S-100 protein the lesion was shown to consist of proliferative histiocytes exhibiting emperipolesis. The histopathological diagnosis was compatible with RDD. The diagnosis of RDD mimicking pachymeningitis is presented, and the previous reported cases of intracranial RDD are reviewed. PMID- 15529793 TI - [A case of the usefulness of MR angiography and diffusion weighted image to evaluate migraine]. AB - A 37-year-old male transported by an ambulance with severe pulsating headache after transient was scintillating scotoma and right hemianopia. He had experienced migraine attacks 2-10 times every year since he was 22 years old. His initial vital signs were stable, the neurological findings while he rested in bed were normal and the head CT examination showed no abnormality. MR angiography revealed narrowing of his cerebral arteries, especially noticeable on the left side. The headache improved after administration of eletriptan hydrobromide. The diffusion weighted image on the next day showed high intensity areas in the left cerebellar hemisphere, vermis, left insular and occipital lobe. The MRA indicated enlargement of the diameter of the cerebral arteries. These findings suggested the usefulness of MRA and diffusion weighted image to evaluate migraine. PMID- 15529794 TI - [Patient selection in stereotaxic neurosurgery for Parkinson's disease: a guide to stereotactic brain operations]. PMID- 15529795 TI - [Clinico-pathology of small skull tumor: giant cell tumor, myofibroma, myofibromatosis, congenital fibromatosis, infantile myofibromatosis, Paget disease of bone]. PMID- 15529796 TI - Lack of randomization in occupational epidemiology. PMID- 15529797 TI - Working population health metrics. AB - This review is concerned with population health metrics that simultaneously consider information on mortality and nonfatal health outcomes as probabilistic measures representing the level of health in a working population. The optimal properties that such a measurement system should have are briefly discussed, and the merits and limitations of health expectancies are compared versus health gap measures. The conceptual basis of these measures are considered, especially with a view to health promotion. The application of worklife expectancies is illustrated with the data obtained from sequential surveys on the work ability status of aging Finnish municipal workers. In conclusion, the developed worklife expectancies are considered to be appropriate indicators for measuring the future duration of occupancy in a given state of work ability, and they should be preferred to alternative measures of population health in the context of occupational health. PMID- 15529798 TI - Cancer incidence of Nordic asphalt workers. AB - OBJECTIVES: The aim of the study was to investigate cancer risk following employment in the asphalt industry. METHODS: Cancer incidence was studied among 22 362 male asphalt workers employed for more than one season in jobs entailing exposure to bitumen (mainly road pavers) in Denmark, Finland, Norway, and Sweden. These workers are part of a European cohort of asphalt workers, for which results on mortality have been reported. The follow-up was almost complete, and reference rates were obtained from national cancer registries. RESULTS: The incidence of cancer was reduced in all four countries [overall standardized incidence ratio (SIR) 0.89, 95% confidence interval (95% CI) 0.86-0.94]. Lung cancer incidence was increased in all four countries, yielding an overall SIR of 1.21 (95% CI 1.07 1.36), but no trend was detected according to time since first employment. No overall increased incidence of bladder cancer was observed, but there was a suggestion of a tendency towards higher risk with longer time since first employment, with a relative risk of 1.85 (95% CI 0.90-3.78) for more than 30 years versus 1-14 years (P-value for trend 0.1). The incidence of lip and stomach cancer was nonsignificantly increased, and the excess was present mainly in Denmark. No increased incidence was detected for other neoplasms, notably malignant melanoma, nonmelanoma skin cancer, and kidney cancer. CONCLUSIONS: The results of this study confirm the pattern of cancer risk detected in the mortality analysis of the European cohort; in addition, they provide suggestive evidence of an excess risk of bladder cancer among asphalt workers. PMID- 15529799 TI - Ovarian cancer and occupational exposure among pulp and paper employees in Norway. AB - OBJECTIVES: A cohort study of female pulp and paper workers in Norway has shown a significantly increased risk of ovarian cancer. Other than the involvement of hormonal and reproductive factors, little is known of the etiology of ovarian cancer. Asbestos and talc are two agents hypothesized to influence the development of the disease. The present study aimed to investigate the association between ovarian cancer and occupational exposure to asbestos, talc, and total dust among Norwegian pulp and paper workers. METHODS: Forty-six cases of ovarian cancer, with four controls each, were included in the study. Occupational exposure was assessed by combining work histories from personnel files, questionnaire information about production processes, and exposure assessments from the mills. To obtain information about possible confounders, cases and controls were invited to participate in a personal interview. RESULTS: The odds ratio for asbestos exposure was 2.02, 95% confidence interval 0.72-5.66. For talc exposure, the odds ratio was 1.10, and for ever exposure to total dust, it was below 1.00. The risk estimates did not essentially differ after adjustment for possible confounding variables. CONCLUSIONS: The results do not confirm an association between exposure to asbestos, talc, and total dust and ovarian cancer among Norwegian pulp and paper workers. However, the odds ratio for asbestos exposure was doubled, and control for established nonoccupational risk factors did not change the estimate. Therefore, the possibility that exposure to substances in the work environment contributes to the elevated risk cannot be rejected. PMID- 15529800 TI - Central nervous system effects of acute organophosphate poisoning in a two-year follow-up. AB - OBJECTIVES: Patients hospitalized for acute organophosphate poisoning in Leon, Nicaragua, were followed for effects on the central nervous system (CNS) over a 2 year period. METHODS: Immediate verbal memory (Rey verbal learning), visuomotor performance (digit symbol), and neuropsychiatric symptoms (Q-16) were assessed for 53 poisoned persons at the time of hospital discharge, 7 weeks postpoisoning, and 2 years postpoisoning, and, at the same time intervals, for 28 persons who had never been poisoned. The poisonings were classified as moderate occupational (31), severe occupational (15), and severe through the oral route (7), representing low, medium, and high exposure, respectively. Longitudinal confounder-adjusted between-category comparisons and longitudinal analyses of variance and covariance were used to assess the effects of the exposure. RESULTS: Immediate verbal learning showed deficits in the high-exposure group, in particular at the time of discharge, but the estimate of the difference when compared with the values of the unexposed was imprecise. Visuomotor performance showed a deficit at 7 weeks in the medium-exposure group, but it had improved after 2 years relative to that of the unexposed, for whom improvement had occurred at 7 weeks and persisted during the 2 years of follow-up, possibly a test-retest effect. Neuropsychiatric symptoms were in excess 2 years after the hospital discharge in the low- and medium-exposure groups and all the groups combined. All the results were imprecise for the small high-exposure group. CONCLUSIONS: Visuomotor performance and possibly short-term verbal memory seem to be affected early after severe acute organophosphate poisoning and recover, either truly or by some compensatory mechanism. Neuropsychiatric symptoms seem to increase after a longer latency period. PMID- 15529801 TI - Exposure to job-stress factors in a national survey in France. AB - OBJECTIVES: Job stress is a growing epidemiologic field in France. The aim of this study was to evaluate the levels of occupational psychosocial risk factors to which the French working population is exposed. It also focused on developing a job-exposure matrix. METHODS: This study used existing French national data on work conditions collected by the French Ministry of Labor in 1991, in which 20 929 workers were interviewed by questionnaire. The items of the questionnaire were retained that represented potential stressors. A principal component analysis was performed to summarize the data in terms of job-stress factors. Four independent variables (gender, age, occupation, and activity) were available, as well as scores for exposure to psychosocial risk factors. The CART (classification and regression tree) segmentation method was used to construct the job-exposure matrix. RESULTS: Fourteen psychosocial factors were identified and interpreted on the basis of the results of the principal component analysis and deepened by experts' judgment. To take into account the well-known difference of distribution in occupations between men and women, a gender-specific matrix was developed. CONCLUSIONS: This study is the first attempt to develop a job exposure matrix in the area of job-stress factors in France. Thus the results have allowed the assessment of exposure to 14 psychosocial factors for all of the 455 categories of the French occupational classification for men and women separately. PMID- 15529802 TI - Computer users' risk factors for developing shoulder, elbow and back symptoms. AB - OBJECTIVES: This prospective study concentrated on determining factors of computer work that predict musculoskeletal symptoms in the shoulder, elbow, and low-back regions. METHODS: A questionnaire on ergonomics, work pauses, work techniques, and psychosocial and work factors was delivered to 5033 office workers at baseline in early 1999 (response rate 69%) and to 3361 respondents at the time of the follow-up in late 2000 (response rate 77%). An increased frequency or intensity of symptoms was the outcome variable, including only nonsymptomatic respondents from the baseline questionnaire (symptom frequency below 8 days within the last 12 months or intensity score below 4 within the last 3 months). RESULTS: In the follow-up, 10%, 18%, and 23% had symptoms more often in the elbow, shoulder, and low back, respectively, and 14%, 20%, and 22% had more intense symptoms. Women were more likely to be afflicted than men in all regions. In the full-fit multivariate logistic regression analysis, little influence on the timing of a rest pause and being disturbed by glare or reflection were significant predictors of shoulder symptoms, screen below eye height was a significant predictor for elbow symptoms, and previous symptoms was a significant predictor for symptoms in all regions. Computer worktime and psychosocial dimensions were not significant predictors. CONCLUSIONS: Influence on work pauses, reduction of glare or reflection, and screen height are important factors in the design of future computer workstations. Since previous symptoms was a significant predictor of recurrent symptoms in all three regions under study, it can be concluded that musculoskeletal symptoms are persistent. PMID- 15529803 TI - Neck and shoulder symptoms and disorders among Danish computer workers. AB - OBJECTIVES: Neck and shoulder pain and disorders were studied among frequent computer users, and the associated effect of mouse and keyboard use was evaluated. METHODS: Technical assistants and machine technicians were followed for 1 year. Questionnaires were sent to 9480 persons (initial response 73%, follow-up response 82%). Computer use information was obtained from the questionnaires. Symptom cases at baseline and follow-up were clinically examined using a standardized clinical protocol. The main outcomes were self-reported pain symptoms in the neck and right shoulder and clinical cases of rotator cuff syndrome, tension neck syndrome, and neck-shoulder pain with pressure tenderness. RESULTS: The prevalence of moderate-to-severe pain in the neck and right shoulder was 4.1% and 3.4%, respectively, and the 1-year incidence for no or minor baseline symptoms was 1.5% and 1.9%, respectively. At baseline, the prevalence rate ratio (PRR) for neck pain was 1.7 [95% confidence interval (95% CI) 1.1-2.6] for mouse use >25 hours/week, that for right shoulder pain increased from 1.6 (95% CI 1.1-2.4) for 15-19 hours/ week to 2.5 (95% CI 1.4-4.3) for >30 hours/week of mouse use, and that for tension neck syndrome increased from 3.5 (95% CI 1.0 12) for 25-29 hours/week to 4.7 (95% CI 1.2-18) for >30 hours/week of mouse use. The relative risk (RR) for new neck pain was 1.8 (95% CI 0.8-3.9) for keyboard use > or = 15 hours/week and increased to 2.4 (95% CI 0.8-6.8) for > or = 30 hours/week. New right-shoulder pain symptoms were associated with mouse use >20 hours/week (RR 1.9, 95% Cl 1.0-3.5, and RR 3.3, 95% CI 1.2-8.9) and with keyboard use >15 hours/week (RR 2.2, 95% CI 1.0-4.9). CONCLUSIONS: Mouse use is associated with an increased risk of moderate-to-severe pain in the neck and right shoulder, and an association with tension neck syndrome is possible. PMID- 15529804 TI - Prospective study of the relationship between musculoskeletal and psychological complaints and electromyographic activity during isometric muscular contractions in a working population. AB - OBJECTIVES: This study focused on determining whether musculoskeletal and psychological complaints reported monthly over a 4-month period predicted muscular activity during and immediately after standardized worktasks and whether muscular activity during and immediately after these tasks predicted changes in complaint severity in the following 12 months. METHODS: Surface electromyography (EMG) was recorded bilaterally from the upper trapezius, middle deltoid, and forearm extensor muscles of 45 postal workers (30 women) during sustained submaximal (25% of peak force) isometric contractions (wrist extension and shoulder abduction). Self-reported health complaints were recorded monthly. Musculoskeletal and psychological complaint-severity indices (MSI and PI, respectively) were computed from complaint-severity scores (intensity score x duration score). The history of complaints over the previous 4 months was included in adjusted regression models to predict muscular activity during and immediately after submaximal contractions. Muscular activity was included in adjusted models to predict changes in the complaint severity over the subsequent 12-month period. RESULTS: A higher MSI predicted a lower EMG level in the trapezius muscle during submaximal contractions (P<0.014), whereas the PI did not predict the level of EMG in any of the muscles studied (P > 0.194). The EMG activity did not predict changes in the complaint severity over the subsequent 12 months. CONCLUSIONS: These findings may support the hypotheses of pain adaptation or the dysfunction of synergistic muscular control in relation to musculoskeletal complaints. However, the findings did not indicate that increased or decreased muscular activity is a risk factor for heightened levels of complaints in the subsequent 12 months. PMID- 15529806 TI - Does respect for embryos entail respect for gametes? AB - Respect for human embryos is often defended on the basis of the potentiality argument: embryos deserve respect because they already possess potentially the features that in adults are fully actualized. Opponents of this argument challenge it by claiming that if embryos should be respected because they are potentially adults, then gametes should be respected because they are potentially embryos. This article rejects this reductio ad absurdum argument by showing that there are two different types of potentiality involved so that the transitivity of potentiality does not hold up in this case. Respect for embryos does not logically entail respect for gametes. PMID- 15529805 TI - Depression in the context of disability and the "right to die". AB - Arguments in favor of legalized assisted suicide often center on issues of personal privacy and freedom of choice over one's body. Many disability advocates assert, however, that autonomy arguments neglect the complex sociopolitical determinants of despair for people with disabilities. Specifically, they argue that social approval of suicide for individuals with irreversible conditions is discriminatory and that relaxing restrictions on assisted suicide would jeopardize, not advance, the freedom of persons with disabilities to direct the lives they choose. This paper examines the idea promoted by some proponents of assisted suicide that it is reasonable to be depressed about one's diminished quality of life in cases of irreversible illness or disability and, therefore, such depression should not call into question the individual's competence to request assistance in dying. The concept of rational depression is defined and examined in the context of: four real-life cases involving individuals with disabilities who requested assistance in dying; a set of criteria commonly applied to decision-making to determine rationality; and research bearing on the emotional status of people with disabilities. It is concluded that although disability is associated with particular socially mediated stressors, there is no theoretical or empirical evidence to indicate that depression and its role in the "right to die" is dynamically different, more natural, or more reasonable for disabled people than for non-disabled people. PMID- 15529807 TI - Respect for embryos and the potentiality argument. PMID- 15529808 TI - [Standard procedures for the diagnosis and management of cases of congenital rubella]. PMID- 15529809 TI - [Probiotics: from Metchnikoff to the current preventive and therapeutic possibilities]. AB - About a century ago, Metchnikoff first hypothesised that some intestinal bacteria "produce compounds useful against a premature ageing". Since then, studies progressed over last century, leading to a remarkable improvement of the knowledge about the role of intestinal micro-organisms. Nowadays a number of different micro-organisms satisfying certain requisites are named probiotics and are produced on a large scale. At present, a rational use of probiotics with preventive and therapeutic purposes has been proposed not only for some gastrointestinal pathologies, such as the infective diarrhea (as recommended by the Italian Society of Pediatric, the Italian Society of Gastroenterology and Hepatology, the Italian Society of Pediatric Infectious Disease, and by International Societies), but also for other pathologic conditions, such as the atopic dermatitis and related affections (as suggested by the American Academy of Dermatology Association guide lines). Moreover, the use of probiotics is going to be extended to other pathologies, such as the inflammatory intestinal and respiratory diseases, and even to the prevention of tooth decay, although the actual preventive and therapeutic effects of probiotics onto these pathologies have to be carefully investigated. It is unknown if the genius Leonardo Da Vinci (1452-1519), stating that "man's life is built up by food" imagined how nutrients can influence human health, besides being essential for life, as it is today increasingly evident. Avoiding an excessive optimism and the thought that an efficacious panacea for all troubles has been found, there are sound reasons to believe that probiotics, and prebiotics as well, can influence human health, through the prevention and therapy of many diseases, although further studies are still requested to fully clarify the mechanisms of action of these micro organisms on each pathology. PMID- 15529810 TI - [Family pediatricians and obstructive sleep disorders]. AB - BREATHING: Problems related to the Obstructive Sleep Disordered Breathing (OSDB) are so many: 1) a noso- graphic setting has still to be defined and this leads to contrasting results concerning the prevalence of the OSDB; 2) the absence of a single pathogenetic trigger which can explain the sudden increase of the number of cases of the OSDB since the 1980's; 3) a poor integration between clinical and diagnostic tests; 4) a not well defined role of the family pediatrician in approaching the OSDB. OBJECTIVES: From the above introduction we can deduce four objectives of the study: 1) verifying the prevalence of the OSDB; 2) studying if an early development of the adenotonsillar tissues can influence the on-set of the OSDB; 3) a better definition of the clinical diagnosis; 4) knowing what decisions the family pediatrician do take as concerns the diagnostic tests and therapy. MATERIAL AND METHOD: This study was carried out on questionnaires completed by 8 family pediatricians which consisted of two parts: the first section regarded the whole population interviewed (2.271 children) and the second more specific was reserved only to the 42 children classified as affected by the OSDB. These 42 children presented at least 3 of the following 4 features during sleep: (1) the parents are worried about the way their child breaths (2) snoring (3) apnea (4) paradoxical rib cage movement in inspiration. RESULTS: The prevalence of the OSDB was 1.8%. However considering how suggested by some authors even those children who snored and also presented oral respiration, the prevalence increased to 10.3%. These values are similar to the international results with a prevalence of 2-3% for the more severe forms defined as Obstructive Sleep Apnea Syndromes (OSAS) and of 8-11% considering all the forms of the OSDB. Grouping these patients according to their ages, it resulted that the highest incidence of the OSDB was in children between 3-5 years. This observation supports the hypothesis that at the base of the OSDB is an early development of the adenotonsillar tissues, thus in constrast which the classical course which identifies the peak of adenotonsillar hypertrophy between 4 and 6 years of age. The frequency of the single signs and symptoms in the various ages permits the improvement of the clinical diagnosis: in particular snoring, oral respiration and tonsillar hypertrophy are less frequent in the first three years of life, while in the older children the percentage of growth inhibition decreased and it becomes more difficult observing paradoxical rib cage movement in inspiration. Concerning the diagnostic tests, the family pediatrician asks only exceptionally specific test during sleep (5% of the patients). Concerning therapy, many were the indications for adenotonsillectomy even during the first three years of age (82% of the patients) proving that the family paediatrician has overcome the old attitude of not indicating operation in the first 4-5 years of age. CONCLUSION: The confirmed high prevalence of the OSDB, the possibility of further improving the clinical diagnosis, the good capacity of the family pediatrician concerns diagnosis and therapy are all factors which favour the direct management of most of the children with adenotonsillar hypertrophy by the family pediatrician. The diagnosis and therapeutic choice can find support in sleep tests when necessary. These tests have to be carried out in a specialized laboratory and the results be interpreted together with the clinical signs and symptoms. Patients who have to be managed by Pediatric sleep laboratory are: 1) children with OSDB due to organic and functional alterations on genetic basis; 2) children in whom adenotonsillectomy presents a high risk such as a severe respiratory insufficiency and the young age of the patient (less 12-18 months of life). PMID- 15529811 TI - [Effects on growth, tolerability and biochemical parameters of two different human milk fortifiers in very low birth-weight newborns]. AB - OBJECTIVES: The aim of the study was to compare a group of very low birth-weight infants feeded with a preterm formula with two other groups feeded with human milk and two different fortifiers. METHODS: 30 preterm newborns with birth-weight < 1.500 g, admitted to the Neonatal Intensive Care Unit (NICU) of Department of Neonatology of Catholic University of Rome were randomized for three different feeding groups: total enteral nutrition with HM fortified with Enfamil Human Milk fortifier or with Eoprotin, compared to a group feeded with Similac 24 preterm formula. Statistical analysis was performed using the two-way analysis of variance (ANOVA). RESULTS: During the study and at the end we found a growth rate for weight, cranial circumference and lenght similar to the fetal standard growth rate in the third trimester of pregnancy in all the three groups. Fortified HM was well tolerated. No pathologic value of the biochemical parameters studied was found. Higher level of serum phosphorus in spite of significantly lower intakes of phosphorus occurred in fortified HM feeded neonates, as if there was a better availability of this nutrient in HM. CONCLUSIONS: This study demonstrates the role of fortified HM as a good alternative to the preterm formula. PMID- 15529812 TI - Infants vertically infected with hepatitis C: a long term longitudinal follow-up. AB - HCV vertically acquired infection is asymptomatic and characterized by a high chronic infection rate; only 9% of HCV infected children shows spontaneous remission. As far as a mild course of the disease has been observed during childhood, we hypothesize that any eventual treatment intervention could be postpone until adolescent age. PMID- 15529813 TI - [Three cases of congenital cutaneous candidosis]. AB - We report three cases of term newborns with Congenital Cutaneous Candidosis (CCC) occured in a ten months period. Two of these showed respiratory distress that require mechanical ventilation and the administration of exogenous surfactant in one case. All the three cases recovered after therapy with fluconazole. Early onset of severe respiratory distress may require intubation and mechanical ventilation, and systemic involvement requires systemic antimichotic therapy. We did not find any predisposing factors of such a rare disease, in spite of the occurrence of three cases in a short period of time. PMID- 15529814 TI - [Sepsis by Candida in VLBW neonates: therapy with liposomal Amphotericin B]. AB - Candida spp. are recently frequent cause of nosocomial sepsis in neonates admitted in NICU, expecially VLBW. Amphotericin B is used in the treament of infections caused by Candida, but heavy side effects, expecially due to renal toxiticy, prevent often its use in the VLBW neonates. We used a new formulation of Amphotericin (AmBisome--NEXSTAR), in the treatment of 17 VLBW neonates affected by sepsis caused by Candida albicans, admitted in our NICU during two years. Twelve neonates survived, one neonate died for NEC and bowel perforation. The treatment was prolonged for 13 days (7-33 days), the total amount of dose was 65 mg/Kg, none side effect was noted. PMID- 15529815 TI - [Spontaneous regression of an adrenal mass detected in utero]. AB - The case of a left adrenal mass detected by ultrasonography in a male fetus at the 37th week of gestation is presented. After an uneventful delivery at the 38th gw, the newborn was sonographically monitored, and during the follow-up clinical evaluations were performed, in order to make a differential diagnosis between an adrenal hemorrhage and a congenital neuroblastoma. VMA and HVA were within normal levels, and MIGB scan was negative. A MR of the the abdomen performed at the age of one month was suggestive for an adrenal hemorrhage in involution. We therefore decided to keep monitoring the mass by ultrasound, avoiding any surgical approach. The sonographic complete disappearance of the disease was documented at the end of the fourth month. PMID- 15529816 TI - [Homozygous hemoglobin-E (Hb-EE) disease]. AB - The Authors report on a 16 year-old girl, of Cambodian descent, who was admitted to the hospital for hematuria. She showed a mild microcytic, hypochromic anemia with a normal iron balance; clinical examination was normal with neither pallor nor icterus nor splenomegaly; electrophoresis of hemoglobin yielded no hemoglobin A, a sligtly increased amount of HbF and a single band with a mobility similar to that of HbA2; the patient showed no evidence of overt increased hemolysis. With the DNA technology a final diagnosis of homozygous hemoglobin E was made. Hemoglobin E is the most common Hb variant among Southeast Asian populations. The Authors discuss on the benign nature of Hb-EE disease, pointing out that the presence of a single HbE gene in combination with that for beta-thalassemia leads generally to a disorder often comparable in severity to that of homozygous beta thalassemia. With the recent migration of a high number of people from the countries, where HbE is extremely frequent, to the Western world (including Italy), this thalassemia syndrome is now a global health problem; therefore its knowledge is an important diagnostic challenge to all the experts involved in the care of thalassemic patients. PMID- 15529817 TI - [Systemic fungal infections in neonates]. PMID- 15529818 TI - The transition to psychosis: risk factors and brain changes. PMID- 15529819 TI - Factors leading to poor physical health in people with psychosis. PMID- 15529820 TI - [Factors leading to bad social outcome in subjects with psychosis]. PMID- 15529821 TI - Trajectories of psychosis: towards a new social biology of schizophrenia. AB - Over the last 2 decades, discourse on the causes of schizophrenia was conducted almost entirely in terms of biological risk factors. This was probably the result of social trends in the research community, and in popular culture, as a wave of techno-optimism promised answers to big human questions in terms of small pixels and even smaller molecules. The human genome project inflated expectations further, and the pharmaceutical industry conspired with the desire of psychiatrists for scientific respectability. 'Social factors', whether at macro societal or locality/family level, came to be seen as 'fall-out' from biological mechanisms, a kind of padding to our understanding of human disease. But changes are in the wind. New understandings of the influence of social factors on the long-term outcome trajectories of psychosis, their potential role in risks associated with migration, and recent findings from genetic high risk studies, are raising fresh questions about social factors and causation. This paper does not argue that the evidence (yet) is strong. But after 2 decades of often crudely articulated dualism, it is time once again for social experience to be integrated with more sophisticated theory development and hypothesis testing in the search for the causes of schizophrenia. PMID- 15529822 TI - [Surveying patients' and family members' satisfaction: the experience of the Mental Health Department in Savona]. AB - AIM: To point out the degree of satisfaction of psychotic patients and their family members in relation to the assistance given by the four Outpatient Mental Health Services in the Savona Department. METHODS: patients with a diagnosis of psychosis were selected among those visited during the period from the 1st of January to the 30th of April 2002. VSSS-54 item, was utilized. RESULTS: The analysis, conducted on 301 patients and 149 family members, highlighted: 1) a substantially positive assessment of the Services 2) the patients and their family members quite agreed on the evaluation of the different areas of satisfaction 3) Strong points: all the operators' human and professional skills 4) request of a better knowledge of the Service's programmes and more information to the public opinion 5) request of more collaboration with family doctors and other specialists 6) The worst criticism: the Service's response to night and holiday emergency. CONCLUSIONS: The patients and their family members reacted positively to the survey, that created more resistance on the part of the operators. The survey is not to be considered the final objective, but the point of departure for a new form of collaboration between the users and the operators. PMID- 15529823 TI - Morale of mental health professionals in Community Mental Health Services of a Northern Italian Province. AB - AIMS: To explore morale of psychiatrists and psychiatric nurses working in Community Mental Health Centres (CMHC) in an Italian Province, and identify influential factors. METHODS: Thirty psychiatrists and 30 nurses working in CMHCs in Modena completed questionnaires on burnout, team identity and job satisfaction. They also answered open questions about different aspects of their work. Answers were subjected to content analysis. Regression analyses were used to identify factors that predicted morale across groups. RESULTS: Psychiatrists had higher scores on emotional exhaustion and depersonalisation. There were no significant differences between the two groups in job satisfaction and job or role perception. Professionals reported positive relationships with patients as the most enjoyable aspects of their job, whilst team conflicts and high workloads were seen as most difficult to cope with. Multivariate analyses showed that being a psychiatrist and perceiving team conflicts as a main cause of pressure in the job predicted higher burnout. CONCLUSIONS: Simple open questions coupled with quantitative measures appear a promising tool to investigate morale of mental health professionals and identify factors determining morale. Research, training and service development should focus on relationship aspects both with patients and within teams to reduce burnout in CMHCs. PMID- 15529825 TI - [The increasing prevalence of pollinosis]. PMID- 15529824 TI - [An observational study of effectiveness in community residential facilities]. AB - AIMS: Evaluation of community residential facilities effectiveness in the Department of Mental Health of Desio (Milan). METHOD: Outcomes in symptoms, disability, family burden and quality of life were evaluated during one year through a longitudinal study, using a pre-test and post test design without control group. RESULTS: Residential care is effective in reducing disability and symptoms, while it is not effective towards family burden. Quality of life is improved in some domains, but not in others (e.g. social and family relationships). CONCLUSIONS: Outcome assessment is feasible in residential facilities, following a multiaxial and multifactorial model. We need to clarify the goals of residential care, focussing on active components of the residential treatment. PMID- 15529826 TI - [Report of the activity of RNSA (National Aero-biological Surveillance Network), 2003]. PMID- 15529827 TI - [Flora, distribution of vegetation and pollen content of the air: significance for allergics]. AB - The wealth of the flora of a given region cannot be seen in its airborne pollen list. Actually, for some plants there is a low probability that their pollens are recorded in the pollen counts (entomogamous plants, in particular). Moreover, the light microscopy reduces the possibilities of determination. In France, The Aerobiological Network of Surveillance (RNSA) retains 92 taxa at different levels (gender, family, group of families), and among them allergenic as well as non allergenic taxa. From the 130 families of the France flora, 63 are taken into account in the pollen studies, 57 taxa being determined at the gender level. The comparison between the surface occupied by the tree species in the French departments of Cote-d'Or and Saone-et-Loire, and the average airborne pollen concentrations showed that usually there is no correspondence between the abundance of a plant and the number of its pollen in the records. So the flora of a region does not give enough information allowing to know the taxonomic composition and the pollen quantities in the air, which is of particular importance for allergic people. PMID- 15529828 TI - [2003, a year of extremes in Burgundy: pollen counts and meteorological influences]. AB - The year 2003 was marked by some particular meterological situations (mild conditions at the end of winter and during spring) which had consequences on the quantity of pollen in the air during the year. These meteorological influences differ from one taxa to another and mostly from one season to another. These quantities very quite different in comparison with the previous years, in particular a notable increase of many winter and spring tree taxa and a decrease of herbaceous taxa, particularly those pollinating in summer. PMID- 15529829 TI - [Pollen content of the Paris air: comparison of the results obtained by two samplers for the year 2003]. AB - During the 2003 season, a second pollen collector has been established in Paris city (Audubon site) to study the representativeness of the initial collector situated on the roof of the Pasteur Institute. The Hygiene Laboratory of Paris followed the pollen counts from the two collectors, during the period going from May to September. Both the samplers are Lanzoni model with a flow rate of 10 l x min(-1). The quantitative results show that no statistical difference exists between the pollen counts obtained from the 2 sites (Mann-Withney test, p > .05). The dominant species are Urticaceae, Poacae and Castenea species for the 2 collectors. Their counts are similar except for Poacae whose results are higher at the Audubon site. The collection period do not take into account the pollen production period of many trees species because of the late installation of the collector on the Audubon site. The results show that the two sites chosen for the study of pollen distribution are comparable. Nevertheless, the comparative study should be maintained on the next year to get more details about the observed differences and to collect the early trees pollen. PMID- 15529830 TI - [How to predict the date of the start of the pollination of a plant from the meteorological data: the example of ragweed at Lyon]. AB - The pollen of Ambrosia, highly allergenic, can be found in high quantities in the French regions of Burgundy, Auvergne and mostly a great part of the Rhone-Alpes region. The aim of this study was to establish some forecast models of the Starting Date of the Pollen season (SDP) of Ambrosia, in order to provide early information to allergists and allergic people. The pollen data came from the Hirst trap set up in Lyon since 1987 by the RNSA, and the meteorological data for the same period were provided by Meteo-France. Two models were used, a sum of daily average temperatures and a multiple regression on all the 10-day period meteorological data. The results were quite accurate, except for 2003 with the temperature sums, probably because of very special meteorological conditions. So the models will have to be updated in this way. PMID- 15529831 TI - [Ragweed pollinosis in the Gard and the Vaucluse: myth or reality? For the Associated Group of Independent Allergologues and Hospital Staff of Nimes]. AB - Thirteen allergists reviewed 18 patients living and working in Vaucluse and Gard who suffered from ragweed from August 15 to October 15 2004. This preliminary study showed how difficult it is to realise a clinical data collection about pollinosis. It is entirely necessary to work together to build the protocol, with a clear and valid definition of inclusion and exclusion criteria. PMID- 15529832 TI - [Description and value of the Microvision system for pollen analysis]. AB - Currently, two techniques of analysis are classically used by the aerobiological networks: the technique of the 12 vertical lines allow to follow the evolution by bi-hourly periods of pollination, but the segmentation of the sample doesn't allow to visualise pollen clouds of short duration; the technique of 2 or 3 horizontal lines allow to follow the daily evolution, but only the daily totals are recorded. The MICROVISION system adapted by the RNSA to the pollen analysis allow to keep the horizontal method, which is faster and continue, and to record automatically in the database the hourly or bi-hourly data. So each pollen is recorded by its coordinates with a precision of 1 micron (1.8 seconds). First, we realised a cartography of the impaction of the pollen grains on the strip, in order to locate the more representative horizontal lines. Secondly, the coupling of the microscope with the Microvision system and the software of data acquiring elaborated by the RNSA allow to propose a more reliable and faster procedure for pollen analysis. PMID- 15529833 TI - Inhibition of graft coronary arteriosclerosis after heart transplantation. AB - Graft coronary arteriosclerosis (GCA) is the leading cause of long-term mortality after heart transplantation (HTx). The goal of this study was to demonstrate that inhibition of immunemediated injury by cyclosporine (CsA) protects the allograft from GCA. ACI-to-Lewis rat allografts were disparate in major and nonmajor histocompatibility loci. Isografts (Lewis-Lewis) were controls. Treatment groups received either olive oil or CsA at 2.5, 5, 10, or 20 mg/kg/day for 3 months. Histology (elastin) and immunohistochemistry using monoclonal antibodies to CD4, CD8, CD45R, RT1B, CD11b/c, CD25, and alpha-actin was performed to examine the epicardial and intramyocardial coronary arteries. Computerized image morphometry was utilized to measure intimal and medial thickness and area. Rats receiving olive oil or CsA at 2.5 mg/kg/day had severe rejection and no graft survival. CsA at 5 mg/kg/day resulted in less severe rejection with significant intimal and medial proliferation (P < 0.001). CsA at 10-20 mg/kg/day paralleled Lewis-Lewis isograft outcomes and inhibited arteriosclerotic vascular changes in the allograft (P < 0.001). Perivascular T-helper cells and macrophages were a characteristic finding with low-dose CsA but rare with higher CsA doses. In this new model of accelerated GCA in rats, immune-mediated antigen-dependent vasculopathy as a result of inadequate immunosuppresion is fundamental in the development of GCA, which appeared equally in epicardial arteries and intramyocardial arterioles. CsA prevents GCA in a dose-dependent fashion in the rat allograft. PMID- 15529834 TI - Influence of transmetatarsal amputation in patients requiring lower extremity distal revascularization. AB - When a transmetatarsal amputation (TMA) is required, successful long-term limb salvage is questioned. We evaluated the influence of TMA on limb salvage in patients undergoing lower extremity revascularization. Patients who had distal bypasses extending to the infrapopliteal arterial tree and adjunctive TMA were retrospectively reviewed. Limb salvage was determined with life-table analysis. Twenty-four patients (29 limbs) were evaluated: 15 male and 9 female. Average age was 64.2 years old. Gangrene was the indication for bypass and TMA in 25 (86.2%) patients. Seven limbs were lost to follow-up. Nine of the remaining 22 limbs required below-knee (8) or above-knee (1) amputations, seven limbs within the first 3 months. In the group of patients who had major amputations within the first 3 months, graft thrombosis was the cause of leg amputation in six (85.7%) cases. No significant predictors of early major amputation were identified. Limb salvage was 62 per cent at 1 year in the TMA group. In comparison, among historical controls requiring distal revascularization and no adjunctive toe or foot amputations, limb salvage was 76.5 per cent (P = NS). Long-term limb salvage is dependent on successful lower extremity revascularization. Requirement for TMA should not influence the decision for limb salvage. PMID- 15529835 TI - The influence of age and gender on the utility of computed tomography to diagnose acute appendicitis. AB - Few studies have examined the effects of age and gender on the utility of diagnostic computed tomography (CT) for appendicitis. We retrospectively studied all adult patients undergoing appendectomy from January 2000 through December 2002 (633 patients). Patients 15-30 years old ("younger") were compared to patients >30 ("older") and further subdivided by gender. CT accuracy and the influence of CT on negative appendectomy (NA) rates for each group were evaluated. CT was associated with a lower NA rate in older patients (23% vs 8%, P = 0.004) but had no effect on NA rate in younger patients (26% vs 20%, P = 0.2). Appendiceal CT had greater sensitivity, negative predictive value, and accuracy in older than in younger patients. When subdivided by gender, CT was associated with lower NA rates for all female patients but had no effect in either age group of men. Although CT may be more accurate in patients older than 30 years, it has the greatest benefit in female patients, where it is associated with lower NA rates in all adult patients. CT should be strongly considered for all female patients before operation for suspected appendicitis. Further study is needed to determine which, if any, male patients benefit from preoperative CT. PMID- 15529836 TI - Accurate diagnosis of infarction of omentum and appendices epiploicae by computed tomography. AB - Segmental infarction of the omentum and epiploic appendages presents with acute abdominal findings that may be confused with a surgical illness. Computed tomography, however, demonstrates a consistent and well-recognized pattern that allows safe, nonoperative treatment. Infarction of omental or mesenteric fat may present clinically as localized peritonitis, mimicking appendicitis, diverticulitis, or cholecystitis. Spontaneous recovery without operation is to be expected if an accurate diagnosis is established. We describe the diagnosis, treatment, and outcome of 15 patients who had infarction of the greater omentum (eight) and epiploic appendage (seven) and presented with localized abdominal pain and tenderness, with six demonstrating regional peritonitis and fever. All underwent CT imaging during their initial evaluation, and 12 of the 15 patients were diagnosed with focal omental or mesenteric fat infarction radiographically and managed nonoperatively. Three patients who had characteristic CT findings nevertheless underwent operation. All patients had complete resolution of their abdominal pain regardless of treatment. The clinical presentation of infarction of the omental or epiploic appendages may be difficult to differentiate from surgical causes of acute abdominal pain. The characteristic findings on computed tomography are diagnostic and allow safe, conservative management in the majority of patients. PMID- 15529837 TI - The efficacy of postoperative oral antibiotics in appendicitis: a randomized prospective double-blinded study. AB - The conventional treatment of acute appendicitis is appendectomy followed by intravenous (IV) antibiotics until intraabdominal infection has resolved. It is controversial as to whether it is efficacious to add a course of oral antibiotics after cessation of IV antibiotics. All consenting patients who presented to Kern Medical Center between October 2000 and June 2003 with acute appendicitis were entered into the study. Perforated/gangrenous appendicitis was equally represented in the two study arms. After appendectomy, and when IV antibiotics were ready to be discontinued, patients were randomized to receive a 7-day outpatient course of either placebo (Group 1) or oral antibiotics (Group 2). Patients were monitored for infectious complications for a minimum of 3 months, and there was no statistical difference (11.5% in Group 1 vs 12.1% in Group 2, P = 0.61). The data suggest that adding a course of outpatient oral antibiotics, after completing a course of IV antibiotics, does not decrease postoperative infectious complications in appendicitis patients. PMID- 15529838 TI - Symptoms before and after laparoscopic cholecystectomy for gallstones. AB - Between 1989 and 1995, 1380 patients underwent laparoscopic cholecystectomy for symptomatic cholelithiasis by a single surgical group at a large private teaching hospital. Thirteen hundred surveys were mailed, and 573 (44.3%) were completed at least 6 months postoperatively. Pain and nonpain symptoms were present preoperatively in 432 (75%) and 457 (80%) patients, respectively. Postoperatively, pain and nonpain symptoms were present in 141 (25%) and 247 (43%) patients, respectively (P < 0.05). All nonpain symptoms were significantly reduced postoperatively except for diarrhea (P < 0.05). Longer duration of pain, age < 40, frequent episodes of pain, postprandial pain, and increased sites of pain preoperatively were all predictive of a higher incidence of persistent postoperative pain (P < 0.05). Persistent nonpain symptoms were more likely if diarrhea, fatty food intolerance, age < 40, or both pain and nonpain symptoms were present preoperatively (P = 0.05) and less likely if only pain symptoms were present preoperatively (P = 0.0001). This series quantifies symptom-specific outcomes for the surgeon. While most symptoms improve, a significant number of pain and nonpain symptoms persist after laparoscopic cholecystectomy. With these data, surgeons can modulate postoperative expectations and advise on the possible persistence of symptoms. PMID- 15529839 TI - Diagnosis of palpable breast masses: ultrasound-guided large core biopsy in a multidisciplinary setting. AB - Cytologic diagnosis of palpable breast masses is an accepted method for diagnosis. However, the high nondiagnostic rate causes repeat biopsy, unnecessary delays, and increased costs. Our purpose is to evaluate the use of ultrasound (US)-guided large-core needle biopsy as part of the minimally invasive multidisciplinary diagnosis of palpable breast masses. We studied 502 consecutive patients with 510 palpable solid breast masses seen and evaluated by a multidisciplinary team. Patients had US-guided core biopsy. Clinical-imaging pathologic correlation (CIPC) was done in all cases. Core biopsy was deemed conclusive if CIPC was congruent and was used to guide definitive management. The median age of our patients was 39 years. Median tumor size was 2.2 cm. Of these cases, 463 (91%) had a conclusive diagnosis on CIPC. Core needle findings on 47 masses were nondefinitive to guide therapy (fibroepithelial lesion, atypical ductal hyperplasia, intraductal papilloma, CIPC). Three cancers were detected in this group on excisional biopsy. In conclusion, US-guided large-core needle biopsy is a sensitive method for diagnosis of palpable breast masses. Multidisciplinary correlation of clinical findings, imaging, and pathology is essential for success. This approach improves use of operating room resources and maximizes patient participation in the decision-making process. PMID- 15529840 TI - Sentinel lymph node mapping of breast cancer: a case-control study of methylene blue tracer compared to isosulfan blue. AB - Isosulfan blue has been traditionally used as a tracer to map the lymphatic system during identification of the sentinel lymph node. However, allergic reactions may be life threatening. We compared the efficacy of methylene blue dye as a tracer for sentinel lymph node biopsy to isosulfan blue dye. In an analysis of 164 cases, there was no clinical or statistically significant difference in the success rate of sentinel node biopsy (P = 0.22), the number of blue sentinel nodes harvested (P = 0.46), the concordance with radioactive sentinel nodes (P = 0.92), or the incidence of metastases (P = 0.87) when methylene blue tracer was compared to isosulfan blue. No adverse reaction to either blue dye was observed. In conclusion, intraparenchymal injection of methylene blue dye is a reliable tracer for the lymphatic system and nodal identification during sentinel node mapping for breast cancer. It is safe, inexpensive, and readily available. PMID- 15529841 TI - Axillary recurrence in DCIs: is axillary lymphadenectomy warranted? AB - Widespread use of screening mammography has resulted in a remarkable increase in the incidence (or detection rate) of ductal carcinoma in situ (DCIS). Axillary lymph node involvement in DCIS is reported to occur at a frequency of 1-12 per cent. Over the past few years, however, there has been increasing emphasis on axillary sampling, limited axillary dissection, and the potential role of sentinel lymph node biopsy. The clinical relevance of axillary lymph node biopsy or dissection remains unanswered. This retrospective analysis was performed on 171 patients who underwent treatment for DCIS at a tertiary care center over a period of 14 years. Clinical and tumor factors were evaluated, and the local, axillary, and systemic recurrence rates were noted. No axillary recurrences from the primary DCIS diagnosis were noted in the entire group of 171 patients. During a mean follow-up of 70 months, 10 patients (6%) developed recurrence in the ipsilateral breast. Six of these recurrences were in the form of DCIS, whereas, four recurred as invasive cancers. Nine patients developed a new primary (seven DCIS and two invasive) in the same breast but in a different quadrant. Two patients with ipsilateral invasive disease also developed systemic disease and eventually died of disease. During the same period, 10 patients (6%) developed DCIS, and seven patients (4%) developed invasive cancer in the contralateral breast. The data show that the risk of axillary recurrence in pure DCIS is, at most, extremely low and support the position that nodal sampling or dissection is unwarranted. PMID- 15529842 TI - Lymphatic mapping improves staging and reduces morbidity in women undergoing total mastectomy for breast carcinoma. AB - Lymphatic mapping (LM) and sentinel lymph node biopsy (SLNB) have become widely accepted in the setting of breast conservation surgery. We hypothesized that LM can be extended to women undergoing total mastectomy, being technically feasible, yielding highly accurate and sensitive results, improving axillary staging, and reducing postoperative morbidity. Between 1995 and 2003, 99 women (mean age 59 years, range 34-87) underwent 100 mastectomies with LM using blue dye alone. Fifty-nine operations (60%) were followed by a completion axillary lymph node dissection (ALND). Ninety per cent of patients had invasive carcinoma; 10 per cent had in situ carcinoma. Mean tumor size was 2.5 cm (range 0.3-8 cm). One hundred fifty-nine sentinel nodes (SNs) (mean 1.65, range 1-5) were successfully identified in 96 (96%) axillae. Twenty-five (25%) sentinel nodes revealed nodal metastases. Five of 25 (20%) SNs had micrometasteses. Three patients had a false negative SN, yielding a sensitivity of 91 per cent. The accuracy of LM was 97 per cent. No patient who underwent SLNB alone developed lymphedema, axillary seroma formation, infection, or restricted arm movement. This was contrasted with patients undergoing ALND, where 10 (16%) developed lymphedema and 2 (3%) developed an infection. Ten (25%) patients developed axillary paresthesias after SNB compared with 47 (78%) patients after ALND (P < 0.0001). LM in the setting of mastectomy is accurate and sensitive. This technique improves axillary staging and decreases morbidity. Patients who are not candidates for breast conservation should be offered LM and SLNB at the time of mastectomy. PMID- 15529844 TI - Rhabdomyolysis after penetrating trauma. AB - Rhabdomyolysis (RHAB) is a known complication following blunt trauma. RHAB after penetrating trauma has not been studied. The objective of this study was to evaluate the incidence, risk factors, and complications of RHAB following penetrating trauma. Over a 5-year period, penetrating trauma patients admitted to our intensive care unit (ICU) were studied. Significant RHAB was defined as a CK level of 5000 U/L or higher. There were 873 patients (29 +/- 12 years old, 92% male), of whom 767 (88%) had abnormal CK levels (range 520-165,943 U/L), and 111 patients (13%) developed significant RHAB. Victims of penetrating trauma who sustain vascular and severe extremity injury are at a sixfold increased risk to develop significant RHAB. Patients with significant RHAB had a higher rate of renal failure (23% vs 7%, P < 0.0001) and longer ICU stay (15 +/- 26 days vs 8 +/ 12 days, P < 0.0001). CK elevations and significant RHAB are common after penetrating trauma. Patients who sustain vascular and severe extremity injury as a result of their penetrating wounds are at high risk to develop significant RHAB, resulting in renal failure and prolonged ICU stay. Therefore, critically injured penetrating trauma patients should be routinely screened with CK levels. PMID- 15529843 TI - The Santa Monica crash: an urban multicasualty event. AB - Mass casualty events provide dramatic challenges for trauma centers and trauma systems. We analyzed the management of victims and assessed the response of the UCLA Healthcare System to the Santa Monica multicasualty event of July 16, 2003, when an elderly man drove his car through a crowded outdoor market and injured 73 people, 10 of whom died (eight at the scene). Of the victims, 26 were treated at UCLA (n = 15) and Santa Monica (n = 11) Medical Centers. Fourteen patients (54%) were female; average age was 41.9 years (range 7 months to 88 years). Fifteen patients were treated in the ER only, and 11 patients required admission. Of the latter, 10 (91%) had multisystem injuries, most commonly musculoskeletal, which occurred in nine patients (82%). Seven patients required immediate operations (orthopedic in six and a pericardial window in one). Three patients required delayed operations (orthopedic and plastic surgery). Most surgical and medical specialties were needed in consultation. Average LOH was 11.8 (range 2-23) days. Mean ISS was 21.2 (range 1-75). There were six complications (three early and three late) and one death from head injury. Seven patients (64%) required rehabilitation. We conclude that mass casualty victims have multisystem injuries of variable severity, which underscores the importance of trauma centers and trauma systems. The large trauma scene and particular need for orthopedic services were notable features of this event. PMID- 15529845 TI - Thromboprophylaxis does not protect severely injured patients against pulmonary embolism. AB - The existing evidence on the effectiveness of thromboprophylaxis after trauma is conflicting. Although prophylaxis with heparin and/or sequential compression devices is practiced widely, many studies failed to document a clear benefit. A recent meta-analysis suggests that prophylaxis does not reduce posttraumatic deep venous thrombosis rates compared to no prophylaxis. The objective of this prospective study is to examine if the use of thromboprophylaxis prevents posttraumatic pulmonary embolism (PE). Sixty-four critically injured patients with clinical evidence of PE were studied by computed tomographic pulmonary angiography and/or conventional pulmonary angiography. PE was diagnosed in 24 (37.5%) patients. Patients with PE were similar to patients without PE with regard to demographics, injury type and severity, operations, and mortality. Thromboprophylaxis was used with equal frequency between PE and no-PE patients (71% vs 80%, P = 0.4). The type of prophylaxis used was similar between patients with PE (17% heparin, 71% sequential compression devices, 17% combination) and patients without PE (32%, 57%, and 10%, respectively; P = 0.16, 0.28, 0.69, respectively). Current methods of posttraumatic thromboprophylaxis may be inadequate. Practices from nontrauma populations have been erroneously extrapolated to the unique trauma population. To reduce the rate of PE after trauma, new methods of thromboprophylaxis should be considered. PMID- 15529846 TI - Abdominal approach to chronic diaphragmatic hernias: is it safe? AB - Standard teaching has been to approach chronic diaphragmatic hernias (CDH) via a thoracotomy. It has been our experience that CDH can be safely approached via an abdominal incision. The objective of this study was to evaluate the outcome of patients undergoing the transabdominal approach for repair of CDH and comparing the outcome with that of patients undergoing a transthoracic (TT) approach. This is a retrospective chart review and was performed of patients with CDH secondary to trauma. Patient demographics, presenting symptoms, operative approach, and complications were collected. Patients were stratified by the surgical approach, TA versus TT. The endpoints of analysis were need for second incision, intraoperative and postoperative complications (enterotomies, pneumonia), need for a chest tube, mechanical ventilation postoperatively, and ICU and hospital days. Twenty-eight patients with CDH repairs performed between Jan 1993 and Dec 2002 were identified. Nineteen patients were in the TA group, and nine were in the TT group. Patients in the TA group had a higher incidence of emergent surgery (68% vs 11%, P = 0.005) and had a lower incidence of postoperative pneumonia (0% vs 33%, P = 0.009). No case of enteric injury from lysis of adhesions in the chest was identified. The need for a second incision (11%), the mortality (11%), ICU stay, and hospital stay were the same between the two groups. It appears that repair of CDH can be performed safely through an abdominal approach. PMID- 15529847 TI - Hand-assisted laparoscopic donor nephrectomy is safe and results in increased kidney donation. AB - The impact of hand-assisted laparoscopic donor nephrectomy on kidney allograft function, perioperative complications, and organ supply was evaluated by retrospective analysis of 41 hand-assisted laparoscopic donor nephrectomy patients and their recipients between January and October 2003. Serum creatinine at discharge, length of stay, estimated blood loss, operative time, and perioperative complications were analyzed. The mean values for laparoscopic donors and their recipients were 1.2 +/- 0.3 and 1.3 +/- 0.8 mg/dL for creatinine, 3.3 +/- 0.8 and 6.7 +/- 3 days for length of stay, and 110.4 +/- 76.9 and 111.6 +/- 56 mL for estimated blood loss, respectively. No major complications occurred in the laparoscopic donors. The number of living kidney donors increased by 94% compared to the mean of the previous 4 years following implementation of the laparoscopic program. Hand-assisted laparoscopic donor nephrectomy is safe, results in excellent allograft function, and significantly increases donation. PMID- 15529848 TI - Laparoscopic bile duct injuries: management at a tertiary liver center. AB - Bile duct injury is a rare but morbid complication of laparoscopic cholecystectomy (LC). This study was undertaken to evaluate the management of 20 patients with bile duct injuries during LC who were referred to a tertiary center with expertise in hepatobiliary surgery and liver transplantation. Sixteen (80%) were female. Mean age was 44 (range 13-70) years. Half of the injuries were distal (Bismuth I), and nearly half were diagnosed at LC. Reoperative repair was attempted in 30 per cent. Mean interval between injury and operation was 6.55 months (range 0 to 36 months). Eighteen patients underwent Roux-en-Y hepaticojejunostomy (HJ). Of the two patients who did not undergo HJ (both Bismuth I), one was treated with transhepatic cholangiography only, and one died of multiorgan failure. There were four minor complications and one late reoperation for stricture. We conclude that bile duct injury after LC is successfully managed in a tertiary center by a hepatobiliary-liver transplant team. Principles of management include anatomic definition of injury, control of sepsis, staged approach involving interventional radiology, and operative techniques refined in liver transplantation including magnification, fine sutures, selective use of internal stent, and liver biopsy. PMID- 15529849 TI - Laparoscopic gastrojejunostomy for gastric outlet obstruction in pancreatic cancer. AB - Up to 20 per cent of patients with pancreatic cancer develop gastric outlet obstruction. Traditionally, these patients have been managed with an open gastrojejunostomy. Laparoscopic gastrojejunostomy may now be a preferable approach. We conducted a retrospective review of nine patients who underwent laparoscopic gastrojejunostomy in 2001-2004. All nine patients had unresectable pancreatic cancer. There were six men and three women. Median age was 66 years (range 36-87). Two patients had prior laparotomies for attempted resection. Four patients had previously placed duodenal stents that failed. Four others had undergone unsuccessful attempts of duodenal stenting. Median operating time was 116 minutes (range 75-300). There were no intraoperative complications or conversions to open procedure. Median time to postoperative oral intake was 4 days (range 3-6), and median postoperative length of stay was 7 days (range 5 18). Eight of our nine patients were palliated successfully using this technique. There were no complications or deaths related to the operation. All patients were discharged from the hospital. Six patients have since died, with a median postoperative survival of 2.5 months (range 1.5-8). Laparoscopic gastrojejunostomy provides safe and effective palliation of gastric outlet obstruction in patients with unresectable pancreatic cancer. This approach allows for rapid palliation in a group of patients with a very limited survival. PMID- 15529850 TI - Application of robotics in general surgery: initial experience. AB - Robotic surgery was recently approved for clinical use in general abdominal surgery. The aim of this study was to review our experience with the da Vinci surgical system during laparoscopic general surgical procedures. Eighteen patients underwent robotically assisted laparoscopic abdominal surgery between June 2002 and March 2003. Main outcome measures were operative time, room setup time, robotic arm-positioning and surgical time, blood loss, conversion to laparoscopy, length of stay, and morbidity. The types of robotically assisted laparoscopic procedures were excision of gastric leiomyoma (n = 1), Heller myotomy (n = 1), cholecystectomy (n = 2), gastric banding (n = 2), Nissen fundoplication (n = 4), and gastric bypass (n = 8). The mean room setup time was 63 +/- 14 minutes, and the mean robotic arm-positioning time was 16 +/- 7 minutes. Conversion to laparoscopy occurred in two (11%) of 18 cases because of equipment difficulty (n = 1) and technical difficulty (n = 1). Estimated blood loss was 91 +/- 71 mL. The mean operative time was 156 +/- 42 minutes, and the robotic operative time was 27% of the total operative time. The mean length of hospital stay was 2.2 +/- 1.5 days. There was one postoperative wound infection and one anastomotic stricture. Robotically assisted laparoscopic abdominal surgery is feasible and safe; however, the theoretical advantages of the da Vinci surgical system were not clinically apparent. PMID- 15529851 TI - Radiological studies after laparoscopic Roux-en-Y gastric bypass: routine or selective? AB - Early detection of complications after laparoscopic Roux-en-Y gastric bypass (LRYGB) can be difficult because of the subtle clinical findings in obese patients. Consequently, routine postoperative upper gastrointestinal contrast studies (UGI) have been advocated for detection of leak from the gastrojejunostomy. The medical records of 368 consecutive patients undergoing LRYGB were analyzed to determine the efficacy of selective use of radiological studies after LRYGB. Forty-one patients (11%) developed signs suggestive of complications. Of the 41 symptomatic patients, two were explored urgently, 39 (10%) had radiological studies, and 16 of them (41%) were diagnosed with postoperative complications. Overall morbidity of the series was 4.8 per cent. Four patients (1.1%) developed a leak from the gastrojejunostomy and were correctly diagnosed by computerized tomography (CT). The sensitivity and specificity of CT in determining leak was 100 per cent, with positive and negative predictive value of 100 per cent. The mortality of the series was 0 per cent. No radiologic studies were performed in asymptomatic patients, and no complications developed in these patients. Our results show that selective radiological evaluation in patients with suspected complications after LRYGB is safe. High sensitivity makes CT the test of choice in patients with suspected complication after LRYGB. Routine radiological studies are not warranted. PMID- 15529852 TI - Colonoscopic perforation and bleeding of the colon can be treated safely without surgery. AB - The incidence of colonoscopic perforation of colon is about 0.3 per cent. The incidence of colonoscopic bleeding is about 0.6 per cent. Many of those patients undergo unnecessary operations. In order to assess the outcome of nonoperative management of those patients with postcolonoscopic perforation and bleeding, the records of 5120 patients who underwent colonoscopies from September 1, 1988 to June 30, 2003 were retrospectively reviewed with attention paid to colonoscopic perforation and bleeding. Their symptoms, management, and outcome were reviewed. There were 2765 male and 2355 female patients. Ages ranged from 9 to 91 years. A total of 1902 patients (37.1%) had polyps removed. Six patients (0.1%) had colonoscopic perforation. All of them presented with abdominal pain 1 to 4 days after colonoscopic polypectomy. All had subphrenic free air or subcutaneous emphysema on the radiogram. All were treated nonoperatively with nothing by mouth and intravenous fluids and antibiotics in the hospital and recovered uneventfully. Six patients (0.1%) had colonic bleeding that occurred 1 to 14 days after colonoscopic polypectomy. All of them were managed by repeat colonoscopy with injection of epinephrine. All recovered without further bleeding. Therefore, postcolonoscopic perforation and bleeding can be treated nonoperatively. It is safe and cost effective. The mortality and morbidity are very low. PMID- 15529853 TI - Endorectal flap advancement repair is an effective treatment for selected patients with anorectal fistulas. AB - Although the majority of anorectal fistulas may be treated with fistulotomy, patients with higher-lying fistulas or women with anterior fistulas in whom simple fistulotomy may threaten continence present a management dilemma. Endorectal flap advancement repair is one potential solution. Chart review was performed of consecutive patients receiving endorectal advancement flaps during a 5-year period (1999-2003). Twenty-nine patients received endorectal advancement flap repair. In 21 (72%) patients, an etiology of the fistula could not be clearly identified, and they were considered to be cryptoglandular in origin. Eight (24%) patients had pathology presumed to be related to prior obstetrical injury. There were 18 (62%) women, and the majority (72%) had anterior fistulas and flaps. Primary success was defined as resolution of symptoms for at least a 3 month period without an additional surgical procedure. If resolution of symptoms after flap placement was achieved after an additional surgical procedure, it was considered a secondary success. Primary success was achieved in 20 (69%) patients, and a secondary success in four (44%) of the remaining nine patients, for a total success of 83 per cent. Anal advancement flaps demonstrate a reasonable success rate when utilized in this group of patients with challenging anorectal fistulas. PMID- 15529854 TI - What proportion of patients with an ostomy (for diverticulitis) get reversed? AB - A common operation for patients with complicated sigmoid diverticulitis is resection and placement of an ostomy (Hartmann procedure). This population-based study examines that proportion of ostomates who undergo reversal. In the California inpatient file, patients admitted for acute diverticulitis in 1995 were identified, including a subset that had surgical resection. Data regarding receipt of ostomy were obtained (4-year follow-up). Demographics and clinical data (procedure, ostomy reversal, time to reversal, comorbidity score, and complications) were collected. In 1995, 11,582 admissions for diverticulitis occurred in California. Of these, 24.2 per cent (n = 2808) underwent surgery at admission; 88.9 per cent were sigmoid/left colectomies; and 41.7 per cent had a Hartmann procedure. Patients with ostomies were older (P = 0.0004) and male (P = 0.03). Median comobidity score was the same for patients with or without an ostomy. Of the 1176 patients who had the Hartmann procedure, 65 per cent underwent reversal (mean 143 days). A larger proportion of men than women had their ostomies reversed (74.5% vs 55.9%, respectively, P < 0.0001). Median comorbidity scores for both groups were low, 0 for those reversed and 1 for nonreversed. Our study shows that although the majority of patients had their ostomies reversed, over 35 per cent did not at 4-year follow-up. Further studies are required to evaluate how this rate may be improved. PMID- 15529855 TI - Diverticulitis: truly minimally invasive management. AB - The purpose of this study is to evaluate the treatment of patients with acute diverticulitis in the inpatient setting using minimal intervention. This was a retrospective study of 75 patients admitted over a 3-year period with acute diverticulitis as evidenced by computed tomography (CT) and clinical scenario. Of the patients enrolled, 24 (32%) had abscesses identified on their initial CT scan. An additional four patients had abscesses noted on a subsequent CT scan obtained because of lack of complete improvement with medical management, thus raising the total number of abscesses to 28 (37%). Of the patients with abscesses, 10 (36%) underwent drainage using a CT-guided percutaneous or ultrasound-guided transrectal approach an average of 6 days after admission. Of the 75 patients, five (7%) required operative intervention during the initial hospitalization for failure of medical management, two (40%) of whom had abscesses on presentation. The overall median length of hospitalization was 5 (interquartile range [IQR] 4-9) days, and 18 patients (24%) had recurrences during the study period. Our conservative approach to percutaneous and surgical intervention resulted in relatively low percutaneous drainage, a low operative rate, and a reasonable length of hospitalization and recurrence rate. PMID- 15529856 TI - [Prion diseases of man and animal]. AB - Summarized in the paper are data on the molecular bases of transmissible spongoid encephalopathy (TSE), its etiology, formation mechanism of prion proteins, STE related strains, specific barriers to TSE, its pathogenesis and treatment. PMID- 15529857 TI - [Molecular epidemiology of the ECHO 30 virus in Russia and CIS countries]. AB - Nucleotide sequences of the VP1 genome of 28 ECHO 30 strains isolated in 1998 2002 in Russia and several CIS countries were determined. The EV30 studies strain were divided into 4 groups according to isolation place and time. Group 1 is presented by 2 strains isolated in 1998 in Russia and Byelorussia. Group 2 comprises 17 strains isolated in 1999-2000 in Russia (its southern regions and Stavropol Territory), Ukraine, Georgia, and Azerbaijan. Five strains isolated in 2002 in Russia (Kalmykia) belong to Group 3; and Group 4 has 4 strains isolated in 2002 in Moldova and Russia (Magadan). A frequently changing EV30 subtype was simultaneously detected in extensive territories. PMID- 15529858 TI - [Immunological safety and sensitizing effect of an MB-7-based vaccine against hepatitis A]. AB - The influence of a vaccine based on the MB-7 strain of hepatitis A virus (VP-MB 7) designed at the "Vector" Center of Virology and Biotechnology was studied. VP MB-7 was found to provoke no allergic response and to have an activating effect on the specific and non-specific responses of cell and humoral immunity similar to those evoked by hepatitis A vaccine "Hep-A-in Vac". PMID- 15529859 TI - [Reactogenicity, safety and immunogenicity of a recombinant bivaccine against smallpox and hepatitis B in limited clinical trials]. AB - The reactogenicity of the embryonic live recombinant variola and hepatitis B bivaccine as tablets (Revax-BT) as well as its safety and immunogenicity were evaluated in clinical trials made in volunteers who had previously immunized or not with variola vaccine. A preliminary conclusion was made on a lack of side effects and drug safety in primary vaccination and been revaccination with low and high doses. Primary immunization of volunteers and as bivaccination with high doses stimulated the most pronounced immune response to the vaccine virus versus such effect observed in immunization of volunteers with low vaccine doses. Humoral immune response to HBs was observed in 75% of volunteers of both groups after as bivaccination. Such response was most pronounced in examinees immunized with low vaccine doses versus those who received high bivaccine doses. At the same time, no protective levels of humoral immunity response to HBs Ag were observed in volunteers first vaccinated. PMID- 15529860 TI - [Influence of isolated virion proteins of hepatitis A virus on the behavior of rats]. AB - Influenza A virus M-proteins can cause depression in experimental animals 1 day after administration. HA and NA, on the contrary, activate motor activity in animals under the similar conditions. The effect of viral proteins is dose dependent. PMID- 15529861 TI - [A virological description of serous meningitis in children immunized with vaccine against epidemic parotitis]. AB - The morbidity structure was analyzed in children vaccinated against epidemic parotitis in 1993-2002. Eight children (4 with serous meningitis and 4 with lesions of the salivary glands) underwent virologic and immunologic examinations. The molecular typing of the SH-gene fragment of the parotitis virus showed the process in 7 cases to be provoked by the vaccination strain. Presumedly, progressing vaccine-associated meningitis inhibits antibody formation. The total incidence of vaccine-associated meningitis was shown, according to Saint Petersburg data, to be not high, which testifies to a low reactogenicity of the Russian vaccine strain. PMID- 15529862 TI - [Inhibition of replication of Venezuelan equine encephalomyelitis with polyclonal antibodies to laminin-binding protein]. AB - A study of temporal and quantitative characteristics of inhibition of replication of Venezuelan equine encephalomyelitis (VEE) virus, strain TC-83, in Vero and CPE on PK cells showed purified polyclonal rabbit antibodies to human recombinant laminin-binding protein (LBP) to be able to block completely the development of cytopathic effect (CPE) in such cells, when infected with 10(7) CPE60. The extent of VEE infection inhibition in Vero was in direct proportion to a concentration of specific antibodies within a range of 0.44-3 microg/100 microl. When antibodies were added to Vero cells after they were infected, there was a gradual attenuation of the inhibition effect, which stopped almost completely 9 hours after the antibodies were placed. Inhibition was effective at 4 degrees C and 37 degrees C. A lack of synthesis of viral glycoprotein E2 in Vero cells infected in the presence of antibodies to LBP is an extra argument proving that the VEE replication is inhibited at early infection stages. The data obtained demonstrated the general LBP significance for the penetration of VEE into mammalian cells and the related importance of designing new antiviral drugs against alpha-viral infection, which are based on blocking the mechanism of receptor penetration of the virus into the cell. PMID- 15529863 TI - [Antiviral activity of human recombinant gamma-interferon and recombinant hybrid protein of tumor necrosis factor-alpha-thymosin-alpha1 on models of herpes-virus and cytomegalovirus infection in vitro]. AB - The conducted studies showed a certain efficiency of gene-engineering preparation of IFN-gamma and TNF-T in virus infections caused by herpes simplex virus 2 (HSV 2) and cytomegalovirus (CMV) in cell cultures of human embryo fibroblast (HEF). The drugs have no viricidal action. IFN-gamma, when used according to the treatment scheme in vitro, proved to be more effective versus TMF-T both in HSV-2 and in CMV. It inhibited significantly the HSV-2 reproduction within the dilution range of 1:50 to 1:500. It was also effective, when used in CMV, within the dilution range of 1:5000 and lower, whereas TNF-T was effective within the range of 1:500 and lower as well as in 0.1 multiple infection. A significantly higher effect was ensured when the drugs were used for prevention. In HSV-2, IFN-gamma inhibited the virus reproduction, like in the treatment scheme, within the dilution range of 1:50 to 1:500, whereas TNF-T was effective in the range of 1:50. In CMV, the drugs' effect, when used for prevention, was similar to that observed in the treatment scheme. The highest inhibition values were registered for HSV-2, when it was used 24 hours before infection (IFN-gamma--2.25 Ig, dilution range of 1:50; TNF-2--1.0 Ig, dilution range of 1:50). IFN-gamma and TNF 2 exert a synergic action on different stages of virus reproduction. A reliable additive effect was ensured in prevention made 4 hour before infection by IFN gamma and TNF-T only in experimental CMV infection. PMID- 15529864 TI - [Antiviral activity of different drugs in vitro against viruses of bovine infectious rhinotracheitis and bovine diarrhea]. AB - In vitro experiments studied the antiviral activity of 11 different drugs against viruses of bovine infective rhinotracheitis (BIRT) and bovine viral diarrhea (BVD). The 50% inhibiting concentrations of the test agents were determined in the monolayers of MDBK and KCT cell cultures. Only did phosprenyl show a virucidal activity against BIRT virus. All the tested drugs significantly inhibited the reproduction of BIRT virus in the sensitive MDBK cell cultures. Thus, bromuridin, acyclovir, ribavirin and methisazonum inhibited the virus by > or = 100,000 times; liposomal ribavirin, gossypolum, anandinum, polyprenolum, phosprenyl, by 1000-10,000 times; eracond and argovit, by 100 times. In experiments on BVD virus, the cultured KCT cells displayed the antiviral activity of bromuridin, phosprenil, polyprenolum, methisazonum, acyclovir, gossypolum, argovit, and ribavirin (in two variants), which caused a statistically significant (100-10,000-fold) decrease in the productive activity of this virus. Eracond and anandid proved to be ineffective. PMID- 15529865 TI - [Safety research of a vaccine preparation in hepatitis A derived from the MB-7 rapidly growing strain]. AB - A study was undertaken to prove that a vaccine drug based on the fast-growing MB 7 (MB-7 VD) of hepatitis A virus (HAV) is safe. It was established that MB-7 VD, when once injected to white outbred mice and when 4 times injected to Hartley guineas pigs, did not cause any hematological and biochemical changes in the peripheral blood or in the condition of the central nervous system of experimental animals, which shows that MB-7 VD is free of any toxic properties. PMID- 15529866 TI - Toxic response of endosulfan to breeding and non-breeding female mosquitofish. AB - To investigate if toxic effects by endosulfan, a commonly used insecticide, are dependent on the breeding condition of an organism, both breeding and non breeding female western mosquitofish, Gambusia affinis, were exposed to 0.1, 0.5, 1 ppb of endosulfan. After a 5-week exposure period, we examined physical factors such as ovary weight and anal fin length in fish of both reproductive conditions and the size of thyroid follicles in non-breeders. Breeding female fish exposed to endosulfan did not show any significant changes in physical factors, but non breeding females exposed to endosulfan had a significantly greater ratio of anal fin/ body length and larger thyroid follicles than did control females. These results demonstrate that the response to exposure to environmental contaminants may be dependent upon seasonal reproductive condition. PMID- 15529867 TI - An assessment of Turkish deltaic landscapes regarding environmental conservation and sustainable management--a case study of the Kizilirmak Delta. AB - Deltas, as macro and micro biological systems at high productive levels, are the natural reserve areas of the world. Those in Turkey present the international importance landscapes as to the various ecosystem characteristics and land use patterns. Nevertheless, these areas are currently facing many problems such as biodiversity loss and environmental degradation owing to coastal settlements and pollution factors. This paper describes the factors responsible for the main features and degradation types of the Deltaic landscapes and their impact for future land use. It also deals with : an overview of the Kizilirmak Delta, one of the most important Deltas along the central part of Black Sea Region (Turkey); presents major ecosystem of it; identifies land use pattern and discusses the threats by environmental and human induced disturbances. The most considering feature of this Delta is to show the largest and most significant wetland area of Turkey which has been able to protect its natural beauties on the Black Sea Coasts. A Delta plain of 56000 hectares extends through the area at the north of the Samsun-Sinop highway. The ecological system of the Delta is extremely rich in terms of its biological variety as well as its fauna (especially number of bird species). But, the Delta including terrestrial areas close to the water resources and the vegetation between aquatic and terrestrial systems, has been constantly modified by human activities. Consequently, some considerations for environmental conservation and sustainable management were put forward for the future generations. PMID- 15529868 TI - Isoperoxidases show differing sensitivity to salicylic acid. AB - The effects of salicylic acid (SA) on the activity of total peroxidase and the patterns of isoperoxidases of cultured tobacco cells were investigated. The total peroxidase activity of tobacco cells was inhibited by 70% when the cells were treated with 5 mM SA for one week. The peroxidase activity of tobacco cells is declined by 90% in the presence of 30 mM SA. Moreover, the activity of isoperoxidases C3, A1, and A3 decreased dramatically with increasing SA concentration, while, one of the anodic isoperoxidases, A2, was somewhat resistant to SA treatment. When isoperoxidase C3 was isolated, SA inhibited the activity of purified C3 in a concentration-dependent manner. The IC50 of isoperoxidase C3 was approximately 0.45 mM. However, the inhibition of isoperoxidase C3 activity was removed by the addition of Fe2+ ion. The possible mechanism of inhibition of peroxidase by SA is discussed. PMID- 15529869 TI - Bleaching of chemithermomechanical pulp (CTMP) using environmentally friendly chemicals. AB - Increasing environmental restrictions require a reduction in the use of chlorine containing bleaching agents mainly chlorine gas. There are several approaches to this goal, including changes in the cooking process and the influent treatment. The aim of this study is to improve the physical and optical properties of chemithermomechanical pulp (CTMP) which has low brightness and strong tendency towards aging in chlorine-free bleaching sequences. For this reason, initially 7 different sets of single stage hydrogen peroxide bleaching operations were performed. After the analysis of the data obtained, the optimum bleaching conditions were as: H2O2 ratio on oven dry (o.d) raw material: 2%, NaOH ratio (on o.d raw material): 1.5%, MgSO4 ratio (on o.d raw material) : 0.5%, Na2SiO3 ratio (on o.d raw material) : 3%, EDTA ratio (on o.d raw material) : 0.5%, reaction time: 60 minutes, reaction temperature: 70 degrees C and concentration: 16%. Then, 11 different sets of two and multistage H2O2 (P), NaBH4 (B), and Na2S2O4 (S) bleaching operations were carried out. Optimum bleaching conditions were found in bleaching of BPSP with 4 stage sequences. As a result, CTMP brightness was observed to increase from 47% to 81.37% elrepho while yellowness of bleached pulps decreased from 321 per thousand to 164.01 per thousand elrepho. PMID- 15529870 TI - Effects of basic oxygen furnace slag and inorganic nutrients on the germination of resting cysts of two toxic dinoflagellates. AB - Effects of basic oxygen furnace (BOF) slag, inorganic nutrients and H2S on the germination of resting cysts of two toxic dinoflagellates Alexandrium catenella/tamarense and Gymnodinium catenatum were studied in batch cultures. The germination rate of the test species has increased by 23-25%, when the concentration of NO3--N or H2S in culture medium has increased to 2.0 ppm. At the treatment of enriched NH4+-N and PO43--p, the germination of resting cyst was increased. Nevertheless, the increased range in germination rates was less than those of NO3--N and H2S. When BOF slag in culture medium increased to 50 mg/ml (or 500 g/m2), the cyst germination rate fell to less than 5%. At higher level of concentrations germination was completely inhibited. Adding BOF slag to the culture medium reduced the concentration of inorganic salts and H2S in seawater and sediments, resulting in the inhibition of cyst germination. These findings demonstrate the potential use of BOF slag on the sediments seed bank of red tide organism because it has an ability to inhibit resting cysts germination. PMID- 15529871 TI - Effect of some herbicides used in Nigeria on Rhizobium phaseoli, Azotobacter vinelandii and Bacillus subtilis. AB - The effect of three herbicides namely Agroxone, Atranex 50SC and 2,4-Damine on Azotobacter vinelandii, Rhizobium phaseoli and Bacillus subtilis were studied. These bacteria were isolated from a bean-garden in Lagos. The results revealed that 2, 4-Damine was the most toxic of the three herbicides studied and Azotobacter vinelandii was found to be most sensitive to the herbicides. There was a reduction in LC50 of herbicides with increased number of days. The percentage survival decreased with increased concentration of herbicides and days for Rhizobium phaseoli and Azotobacter vinelandii while an initial reduction in population was followed by increased percentage survival of organisms for Bacillus subtilis. PMID- 15529872 TI - Influence of pretreatment and desiccation on the germination of Laurus nobilis L. seeds. AB - The effects of moist stratification, gibberellic acid (GA3) in combination with cold-moist stratification and desiccation on the germination of bay laurel (Laurus nobilis L.) seeds were studied. It was found that the external fleshy pericarp caused dormancy. There was also embryo dormancy and it was broken in seeds lacking pericarp either warm-moist stratification (20 degrees C day/4 degrees C night) or cold-moist stratification (4 degrees C) for 8 and 10 weeks, respectively. Gibberellic acid had little effect on germination percent and germination value of seeds without pericarp when used alone but significantly increased overall germination performance when used with cold-moist stratification. The viability of seeds dropped when the moisture content of the seeds was reduced by desiccation, and the time required to reach the critical moisture levels in the seeds differed in the different desiccation temperatures. The critical minimum seed moisture content for bay laurel was around 15% and the seeds showed the typical behavior of recalcitrant seeds. PMID- 15529873 TI - Heavy metal accumulation by plant species from a coal mining area in Orissa. AB - An ecological survey was undertaken on metal contamination (Cu, Fe, Al, Cr) of vegetation (tree, shrub and herb) collected from the overburden soil of South Bolanda, Talcher, Orissa. Stem and leaf parts of the trees and shrubs, and whole plant of the herbs were analysed. Concentrations of metals were the maximum in leafy part of trees and shrub samples. Among the various heavy metals studied, Fe concentration in plant parts was the highest and Cu concentration was the lowest. PMID- 15529874 TI - Evaluation of the possible role of five enzymes in the metabolism of IBA in mustard aphid, Lipaphis erysimi (Kalt.). AB - The enzymatic activity of five enzymes viz. Glutathione S-transferases, Esterases, NADH dehydrogenase, NADH oxidase and Glutathione reductase were assessed under the influence of Indole butyric acid (IBA) (400 ppm) in the nymphs (48-52h old) of mustard aphid, Lipaphis erysimi fed on radish plants treated for 13, 25 and 37h. The activity of Glutathione S-transferases, Esterases and NADH dehydrogenase increased compared to that found in the control of the same age group of nymphs and it was concluded that these enzymes might be involved in the metabolism of IBA. The other two enzymes, NADH oxidase and Glutathione reductase showed no significant increase in their activity compared to that in the control of the same age group. It was hypothesized that the latter enzymes do not play any significant role in the metabolism of IBA. PMID- 15529875 TI - Groundwater quality assessment in different land-use areas of Faridabad and Rohtak cities of Haryana using deviation index. AB - Physico-chemical parameters were analysed to evaluate the groundwater quality of the two important cities of Haryana, Faridabad and Rohtak, and the pollution status of groundwater was compared using deviation index (DI). Groundwater of both the cities had high alkalinity, hardness, total dissolved solids (TDS) and electrical conductivity. The groundwater of Faridabad showed low fluoride concentration whereas in 74% of groundwater samples of Rohtak the fluoride levels were high. Land use and waste disposal practices were found to have an important effect on groundwater pollution. PMID- 15529876 TI - Alterations in activities of acid phosphatase, alkaline phosphatase, ATPase and ATP content in response to seasonally varying Pi status in okra (Abelmoschus esculentus). AB - Phosphorus (P) is the second most important macronutrient for plant growth. Plants exhibit numerous physiological and metabolic adaptations in response to seasonal variations in phosphorus content. Activities of acid and alkaline phosphatases, ATPase and ATP content were studied in summer, rainy and winter seasons at two different developmental stages (28 and 58 days after sowing) in Okra. Activities of both acid and alkaline phosphatases increased manifold in winter to cope up with low phosphorus content. ATP content and ATPase activity were high in summer signifying an active metabolic period. Phosphorus deficiency is characterized by low ATP content and ATPase activity (which are in turn partly responsible for a drastic reduction in growth and yield) and enhanced activities of acid and alkaline phosphatases which increase the availability of P in P deficient seasons. PMID- 15529877 TI - Demonstration of sperm head shape abnormality and clastogenic potential of cypermethrin. AB - Adult male Swiss albino mice were administered ip. suspension solution of cypermethrin in 0.15% DMSO at the doses of 30 mg, 60 mg and 90 mg/kg b. wt. daily for 5 days. Another group of animals was injected cyclophosphamide ip. (60 mg/kg b. wt.) in similar manner which served as positive control. Effect of cypermethrin on body and testes weight and sperm head morphology was studied. Clastogenic potential of cypermethrin was studied by using modified Allium test. The cytological changes were studied in the root tip cells of Allium cepa after 3 days treatment with three different concentration of cypermethrin (0.1, 1.0 and 10.0 microg/ml). The results revealed that body weight gain was considerably reduced in higher dose groups, but the testicular weight did not change significantly in any of the cypermethrin treated groups. However, a significant elevation in the number of abnormal shape of sperm head was noticed in higher dose groups as compared to control. It was observed that the abnormality in the shape of sperm head was dose-dependent. The cytological changes in the root tip cells of Allium cepa indicated that cypermethrin is having toxic effects on the root tip cells in the form of stickiness of chromosomes and also affect the mitotic activity. This study suggest that cypermethrin may have the potential to induce adverse effects on sperm head shape morphology of mouse as well as clastogenic effects on root tip cells of Allium cepa. PMID- 15529878 TI - Pollution monitoring with the help of lichen transplant technique at some residential sites of Lucknow City, Uttar Pradesh. AB - Samples of foliose lichen, Dirinaria consimillis (Fr.) Awasthi were transplanted at six residential sites of Lucknow city for monitoring the concentration of Cr, Zn, Pb, Cu and Cd metals at two vertical positions. The concentration of chromium and copper were more at the higher vertical position than the lower one. The maximum zinc was at lower vertical position than the upper one. Lead and cadmium were not detected in the present study. PMID- 15529879 TI - Effect of Pseudomonas aeruginosa (MTCC 1688) on the tissue phosphatases activity on Macrobrachium rosenbergii (De Man). AB - A time course study on the endotoxin toxicity of the gram negative bacteria, Pseudomonas aeruginosa MTCC 1688 on the tissue phosphatases activity on the giant freshwater prawn, Macrobrachium rosenbergii was conducted. The results revealed marked elevation of both acid and alkaline phosphatase activity in the haemolymph and body muscle. The hepatopancreas showed reduced phosphatase activity compared to control. The enzymes, being non-specific in action and particularly the acid phosphatase being of lysosomal origin, their increase in muscle and haemolymph has pathogenic significance in the inoculum treated prawns. PMID- 15529880 TI - Pesticide effects on microbial population and soil enzyme activities during vermicomposting of agricultural waste. AB - The effects of different pesticides (propiconazole, profenofos, pretilachlor) on vermicomposting were evaluated for the adverse effects on soil enzyme activities (dehydrogenase, phosphatase, urease) and total microbial counts (TMC). There were remarkable increase in enzyme activities and TMC in presence of earthworm compared to control (earthworm absent). In comparison to control, the activities of phosphatase, dehydrogenase and urease in presence of vermicompost increased upto 30, 128 and 31.3% respectively; whereas increase of TMC was 71.9%. But, in presence of each of the pesticide, said activities decreased. Maximum inhibition of soil phosphatase activity (46.6%) was observed in presence of propiconazole (100 mg kg(-1)) after 120 days. Profenofos affected the soil dehydrogenase activity in the tune of 47% at 1000 mg kg(-1) concentration after 80 days and thereafter, the extent of toxicity decreased little. Soil urease activity was affected markedly in presence of profenofos and was 62% at 1000 mg kg(-1) level after 80 days. TMC also declined in presence of profenofos and pretilachlor. Increase in TMC was about 71.9% compared to control and the inhibition was more or less 60% when profenofos (1000 mg kg(-1)) was present even after 120 days of treatment. PMID- 15529881 TI - Entomological observations on dengue vector mosquitoes following a suspected outbreak of dengue in certain parts of Nagaland with a note on their susceptibility to insecticides. AB - Three species of Aedes viz., Aedes albopictus, Aedes aegypti and Aedes annandalei were detected from different breeding sources in and around human habitats during entomological study conducted following an outbreak suspected to be of dengue (which occurred during, 1994) in parts of Medziphema PHC area of Nagaland in two different points of time ie., in the year, 1994 and, 2000. The potential dengue vector, Aedes albopictus showed high preponderance by breeding in all types of containers searched with high Breteau Index (BI) value of 85.0 and 72.72 recorded in, 1994 and, 2000 respectively whereas the BI value for other potential vector, Aedes aegypti was recorded low (4.9) in the year, 1994 with a substantial increase (31.81) in, 2000. The change in ecosystem along with the process of urbanization has facilitated the growth of these dengue vector mosquitoes in the area of investigation. Adults of both Aedes aegypti and Aedes albopictus were found to be susceptible to DDT, dieldrin and malathion in insecticide bioassay carried out using WHO test kit. PMID- 15529882 TI - Profiles of acid and alkaline phosphatases in the prawn Macrobrachium malcolmsonii exposed to endosulfan. AB - The present study was performed to determine the toxic effects of endosulfan on the quantitative and qualitative aspects of acid and alkaline phosphatases (ACP and ALP) in Macrobrachium malcolmsonii. Intermoult juvenile prawns were exposed to 32.0 ng/l of endosulfan for a period of 21 days. Samples were taken from the hemolymph, hepatopancreas, gills and muscle of test prawns on the 21st day. The content of ACP and ALP in the hepatopancreas of test prawns were found to be higher in comparison to respective controls. The levels of these enzymes in the hemolymph, gills and muscle of test prawns were found to be lower than in the same tissues of controls. In non-denaturing PAGE, phosphatases appeared as white bands. The intensity of white bands in the hepatopancreas of test prawns were found to be higher when compared to controls. In the hemolymph, gills and muscle of test prawns, the intensity of white bands were found to be lower in comparison to controls. The results of the present study suggest that endosulfan affects the quantity and quality of ACP and ALP in the tissues of M. malcolmsonii. PMID- 15529883 TI - Seasonal changes in population of some selected earthworm species and soil nutrients in cultivated agroecosystem. AB - Studies were conducted on population dynamics of Metaphire posthuma, Lampito mauritii and Dichogaster bolaui in cultivated pedoecosystem of desert region of Rajasthan. The populations of aclitellate and clitellate M. posthuma, L. mauritii and D. bolaui were maximum in rainy season and minimum in summer season. The abundant population of these worms were found during the months of July to October. The species M. posthuma breed throughout the year except in extreme summer but L. mauritii breed twice in a year in the field. However, D. bolaui breed once in a year. Among these earthworm species D. bolaui with relative density of 44.2% was the most dominant species in cultivated land while L. mauritii and M. posthuma contributed 33.3% and 22.5% of density, respectively. The total population of earthworms showed significant positive correlation with different soil nutrients. PMID- 15529884 TI - Annelids of a west African estuarine system. AB - Annelids collected from eight stations in the western part of Lagos Lagoon between May, 1996 and February, 1998 were reported. Rainfall, salinity and sediment type influenced distribution of annelids, which were either dry season species, wet season species or euryhaline species. A total of fifty-five species were collected, thirty-one not recorded in earlier studies. Pollution indicators, Polydora sp., Capitella capitata, Nereis sp. and Tubifex sp. were collected in both dry and wet seasons. PMID- 15529885 TI - Noncontingent reinforcement, alternative reinforcement, and the matching law: a laboratory demonstration. AB - Basic researchers, but not most applied researchers, have assumed that the behavior-decelerating effects of noncontingent reinforcement result at least partly from adventitious reinforcement of competing behaviors. The literature contains only sketchy evidence of these effects because few noncontingent reinforcement studies measure alternative behaviors. A laboratory model is presented in which concurrent schedules of contingent reinforcement were used to establish a "target" and an "alternative" behavior. Imposing noncontingent reinforcement decreased target behavior rates and increased alternative behavior rates, outcomes that were well described by the standard quantitative account of alternative reinforcement, the generalized matching law. These results suggest that adventitious reinforcement of alternative behaviors can occur during noncontingent reinforcement interventions, although the range of conditions under which this occurs remains to be determined in future studies. As an adjunct to applied studies, laboratory models permit easy measurement of alternative behaviors and parametric manipulations needed to answer many research questions. PMID- 15529886 TI - Effects of modeling versus instructions on sensitivity to reinforcement schedules. AB - This study examined the effects of modeling versus instructions on the choices of 3 typically developing children and 3 children with attention deficit hyperactivity disorder (ADHD) whose academic responding showed insensitivity to reinforcement schedules. During baseline, students chose between successively presented pairs of mathematics problems associated with different variable interval schedules of reinforcement. After responding proved insensitive to the schedules, sessions were preceded by either instructions or modeling, counterbalanced across students in a multiple baseline design across subjects. During the instruction condition, students were told how to distribute responding to earn the most reinforcers. During the modeling condition, students observed the experimenter performing the task while describing her distribution of responding to obtain the most reinforcers. Once responding approximated obtained reinforcement under either condition, the schedules of reinforcement were changed, and neither instruction nor modeling was provided. Both instruction and modeling interventions quickly produced patterns of response allocation that approximated obtained rates of reinforcement, but responding established with modeling was more sensitive to subsequent changes in the reinforcement schedules than responding established with instructions. Results were similar for students with and without ADHD. PMID- 15529887 TI - Use of a short-term inpatient model to evaluate aberrant behavior: outcome data summaries from 1996 to 2001. AB - Previous outcome studies have provided descriptions of functional analyses conducted in outpatient clinics (Derby et al., 1992), long-term inpatient programs (Iwata, Pace, et al., 1994), and home environments (Wacker et al., 1998). This study provides a description of 138 children and adults with and without developmental disabilities who were evaluated and treated for aberrant behaviors on a short-term inpatient unit. The results indicated that the functional analyses conducted during a short-term inpatient evaluation were successful for 96% of the participants in identifying maintaining reinforcers of aberrant behavior and leading to an 80% or greater reduction in aberrant behavior for 76% of the participants in an average of 10 days. PMID- 15529888 TI - Gross-motor skill acquisition by preschool dance students under self-instruction procedures. AB - The effects of two training procedures--(a) modeling and praise and (b) self instruction, modeling, and praise--on complex gross-motor chain acquisition for preschool dance class students were evaluated. Six girls participated in the study. A multiple baseline design across six gross-motor chains with a secondary group comparison for treatment order effects was used. Both procedures were effective in teaching the gross-motor chains. Nevertheless, for 4 of the 6 participants, the self-instruction procedure produced a faster acquisition rate in at least two of the three comparable pairs of gross-motor chains. Furthermore, very early in gross-motor chain acquisition, for five of the six gross-motor chains, the self-instruction condition was associated with faster response acquisition. PMID- 15529889 TI - A comparison of dense-to-lean and fixed lean schedules of alternative reinforcement and extinction. AB - Behavior-reduction interventions typically employ dense schedules of alternative reinforcement in conjunction with operant extinction for problem behavior. After problem behavior is reduced in the initial treatment stages, schedule thinning is routinely conducted to make the intervention more practical in natural environments. In the current investigation, two methods for thinning alternative reinforcement schedules were compared for 3 clients who exhibited severe problem behavior. In the dense-to-lean (DTL) condition, reinforcement was delivered on relatively dense schedules (using noncontingent reinforcement for 1 participant and functional communication training for 2 participants), followed by systematic schedule thinning to progressively leaner schedules. During the fixed lean (FL) condition, reinforcement was delivered on lean schedules (equivalent to the terminal schedule of the DTL condition). The FL condition produced a quicker attainment of individual treatment goals for 2 of the 3 participants. The results are discussed in terms of the potential utility of using relatively lean schedules at treatment outset. PMID- 15529890 TI - An analysis of initial acquisition and maintenance of sight words following picture matching and copy cover, and compare teaching methods. AB - This study compared the copy, cover, and compare method to a picture-word matching method for teaching sight word recognition. Participants were 5 kindergarten students with less than preprimer sight word vocabularies who were enrolled in a public school in the Pacific Northwest. A multielement design was used to evaluate the effects of the two interventions. Outcomes suggested that sight words taught using the copy, cover, and compare method resulted in better maintenance of word recognition when compared to the picture-matching intervention. Benefits to students and the practicality of employing the word level teaching methods are discussed. PMID- 15529891 TI - Effects of a driver enforcement program on yielding to pedestrians. AB - A driver-yielding enforcement program that included decoy pedestrians, feedback flyers, written and verbal warnings, and saturation enforcement for a 2-week period was evaluated in the city of Miami Beach using a multiple baseline design. During baseline, data were collected at crosswalks along two major corridors. Treatment was introduced first at selected crosswalks without traffic signals along one corridor. A week later, enforcement was shifted to crosswalks along the second corridor. Results indicated that the percentage of drivers yielding to pedestrians increased following the introduction of the enforcement program in each corridor and that these increases were sustained for a period of a year with minimal additional enforcement. The effects also generalized somewhat to untreated crosswalks in both corridors, as well as to crosswalks with traffic signals. PMID- 15529892 TI - Improving day-treatment services for adults with severe disabilities: a norm referenced application of outcome management. AB - We evaluated an outcome management program for working with staff to improve the performance of adults with severe disabilities in a congregate day-treatment setting. Initially, observations were conducted of student task involvement and staff distribution of teaching interactions across students in four program sites. Using recent normative data to establish objective goals for student performance, management intervention was warranted in two of the sites. A six step outcome management program was then implemented in the two sites. The program involved defining desired student and staff outcomes, systematic monitoring of the outcomes, staff training, and supportive and corrective feedback. The outcome management program was accompanied by increases in student on-task behavior and staff distribution of teaching interactions in both sites. The increases brought the levels of on-task behavior above the normative average; on-task behavior was maintained above the baseline average for over 1 year in both sites. These results are discussed in terms of the benefits of relying on normative data for objectively evaluating and improving service delivery systems. Discussion of future research needs focuses on applying the outcome management program to other settings and services for people with disabilities. PMID- 15529893 TI - The effects of delayed physical prompts and reinforcement on infant sign language acquisition. AB - Researchers and clinicians have recommended that sign language be taught to typically developing children during their first 2 years of life; however, existing research does not provide adequate information regarding appropriate methods of sign training. We used delayed physical prompting and reinforcement to teach manual signs to 3 children between the ages of 6 and 13 months. Data were collected on the occurrence of prompted and independent signs as well as crying. Sign training was successful in producing independent signing in all 3 children in under 4 hr of training per child. PMID- 15529894 TI - Behavioral momentum in sports: a partial replication with women's basketball. AB - Previous research has applied the behavioral momentum metaphor to men's college basketball. In the current investigation, the relative rate of reinforcement prior to and following adversities (e.g., turnovers, fouls) and periods of time out were examined in a subset of women's college basketball games. PMID- 15529895 TI - Analogue functional analysis of movements associated with tardive dyskinesia. AB - We studied whether movements associated with tardive dyskinesia (TD) served operant functions in 2 men with developmental disabilities. We found that TD related movements occurred more frequently in the alone and attention conditions and less frequently in control and demand conditions. Our findings suggest that TD-related movements may not be maintained by social reinforcers and that decreases in TD movements are possibly a result of engagement in activities that are incompatible with TD movements. PMID- 15529896 TI - Functional analysis and treatment of the bizarre speech of dually diagnosed adults. AB - Nine behavior-analytic studies, each reporting data for a single participant, have shown that bizarre speech can be maintained by social reinforcement. In the current study, we controlled for a possible referral bias in this literature by including nonreferred participants with dual diagnoses. Functional analyses identified attention functions for 2 participants and nonsocial functions for the others. Noncontingent reinforcement decreased the bizarre speech of both participants who displayed attention-maintained bizarre speech. PMID- 15529897 TI - Functional analysis and treatment of inappropriate sexual behavior. AB - The results of a functional analysis showed that inappropriate sexual behaviors exhibited by a 9-year-old boy who had been diagnosed with traumatic brain injury were maintained by positive reinforcement in the form of social attention. An intervention consisting of functional communication training and extinction resulted in reduced levels of inappropriate sexual behaviors. PMID- 15529898 TI - A preliminary analysis of the effects of response cards on student performance and participation in an upper division university course. AB - We evaluated the effects of response cards on student quiz scores and participation in an upper division undergraduate course at a small, private university. Results showed that response cards increased both quiz scores and student participation. In addition, a measure of the social validity of the response-card procedure suggested that students approved of the use of the cards. PMID- 15529899 TI - A comparison of response cost and differential reinforcement of other behavior to reduce disruptive behavior in a preschool classroom. AB - This study investigated the effectiveness of response cost and differential reinforcement of other behavior (DRO) in reducing the disruptive behaviors of 25 children in a preschool classroom. Using an alternating treatments design, disruptive behavior was reduced when the participants earned tokens for the absence of disruptive behavior (DRO) or lost tokens for the occurrence of disruptive behavior (response cost). Initially, DRO was more successful in reducing the number of disruptive behaviors; however, over time, response cost proved to be more effective. PMID- 15529900 TI - Creating tic suppression: comparing the effects of verbal instruction to differential reinforcement. AB - The purpose of this study was to compare two methods designed to produce tic reduction in 4 children with Tourette's syndrome. Specifically, a verbal instruction not to engage in tics was compared to a verbal instruction plus differential reinforcement of zero-rate behavior (DRO). Results showed that the DRO-enhanced procedure yielded greater reductions in tic frequency. PMID- 15529901 TI - Pedometers and brief e-counseling: increasing physical activity for overweight adults. AB - Physical inactivity has emerged as a public health epidemic and is associated with the rising obesity rate. A multiple-treatments reversal design was utilized to test whether pedometer-aided self-monitoring and brief e-counseling could help 3 overweight adults increase their physical activity. Dependent measures were taken for pedometer steps and weight. Two participants approximately doubled their daily step totals and lost a modest amount of weight by study end. PMID- 15529902 TI - Use of newsletters to promote environmental political action: an experimental analysis. AB - The managers of beach businesses were randomly assigned to receive no intervention or two newsletters per week for 6 weeks that presented economic consequences of damage to coastal water quality and how to contact politicians. The percentage of proenvironmental political contacts was significantly greater in the experimental (newsletter) group than in the control group. Logistical regression showed that contacts in the newsletter group were more than five times that of controls after adjusting for history of contacts and predisposition to take such action. Results suggest that the intervention can promote political action that could influence environmental policies and human health. PMID- 15529903 TI - Fourier phase analysis can be used to objectively analyze real-time myocardial contrast echocardiograms. AB - BACKGROUND: Real-time myocardial contrast echocardiography (MCE) is increasingly used to assess myocardial perfusion. However, objective methods for evaluating MCE are not yet widely available. We sought to validate the ability of Fourier analysis applied to MCE to assess serial changes in microvascular perfusion during coronary occlusion and reperfusion. METHODS: Six pigs underwent 45 min of left anterior descending coronary artery (LAD) occlusion followed by 120 min of reperfusion. Real time MCE was performed at baseline, during coronary occlusion, and at 5, 30, 60 and 120 min of reperfusion. Signal intensities from replenishment curves were fitted to an exponential function to obtain plateau SI (A) and the rate of SI rise (b). MCE images were mathematically transformed using a first-harmonic Fourier algorithm displaying the sequence of myocardial intensity changes as phase angles in parametric images. The phase angle difference (PD) of posterior vs. anterior region was calculated as an index of myocardial opacification heterogeneity and compared to MCE index of myocardial blood flow A x b. RESULTS: After initial hyperemia, a progressive reduction in flow was observed during reperfusion. During LAD occlusion signal intensities were significantly reduced in anterior regions (A x b = 0.02+/-0.01) compared to baseline (1.2+/-0.34, p < 0.01) defining risk areas and approached higher levels postrecanalization (A x b = 1.48+/-0.6) but gradually decreased during 120 min of reperfusion (A = 0.51+/-0.3, p < 0.01). Similarly, profiles of phase angles in LAD perfusion territorities were consistently modified during reperfusion. The mean PD at baseline was 18 degrees+/-15 degrees. PD decreased during coronary occlusion to -108 degrees+/-38 degrees, increased to 29 degrees+/-19 degrees postrecanalization but decreased to -61 degrees+/-35 degrees after 120 min of reperfusion. PD significantly correlated with A (r = 0.8, p < 0.0001) and b (r = 0.73, p < 0.0001). CONCLUSIONS: The progressive reduction in post-ischemic microvascular perfusion was accurately detected by real-time MCE. Fourier phase imaging is feasible to quantify dynamics of myocardial opacification in a simple and objective format and is a promising approach for the interpretation of contrast echocardiograms. PMID- 15529904 TI - Teddy bear in the heart. AB - In a patient with native aortic valve endocarditis, transoesphageal echocardiography yielded a teddy bear appearance which is not reported so far. A perivalvular abscess (right ear), the superior vena cava in cross section (left ear) and the dilated (post-stenotic) aortic root (face) made up the teddy bear. This was not a cuddlesome toy but an ominous sign. The genesis of perivalvular abscess as well as the role of transoesphageal echocardiography in its diagnosis and treatment are briefly reviewed. PMID- 15529905 TI - Prediction of ductus arteriosus closure by neonatal screening echocardiography. AB - AIMS: We reported the incidence, clinical manifestations and outcome of patent ductus arteriosus (PDA) in full-term infants by screening echocardiography. METHODS AND RESULTS: Total 1230 infants received echocardiographic screening and periodic follow-up. On the third day of life, there were 109 infants with isolated ductus arteriosus aneurysm (DAA), those with persistent patency of the ductus arteriosus (DA) in 26. The DA closed within 48 h in 828 infants, and persisted beyond 48 h in 180. The incidence of PDA in full-term infants was 0.91, 0.83 and 0.66% at 1, 4 and 6 months follow-up, and 0.82, 0.73 and 0.64% if infants with DAA were excluded. Infants with PDA had lower birth body weight (BBW) than those with closed DA (p = 0.02). The DA width on the third day of life ranged from 0.23 to 0.43 cm (0.32+/-0.08 cm) and 0.08 to 0.44 cm (0.16+/-0.06 cm) in infants with or without PDA, respectively, which was significantly different (p < 0.05). The sensitivity, specificity, predictive positive and negative values were 87.5, 92.5, 36.8 and 99% respectively for the cuff-off point at 0.25 cm of DA width. CONCLUSIONS: Risk factors in infants with PDA included lower BBW and larger measured DA width at the third day of life. We predicted the arterial duct would close if DA width <0.25 cm, and suggested follow up earlier if >0.25 cm on the third day of life. PMID- 15529906 TI - Large ascending aortic aneurysm and severe aortic regurgitation in a 7-year-old child with Marfan syndrome and a review of the literature. Marfan syndrome in childhood. AB - A 7-year-old girl was admitted because of dyspnea on exertion and palpitations. Her symptoms had gradually worsened for the last 6 months. She had physical features of the Marfan syndrome. Transthoracic echocardiography showed an ascending aortic aneurysm, severe aortic regurgitation, and mildly dilated left ventricle. Because of marked aortic aneurysm and severe aortic regurgitation, the patient was treated with a beta-blocker and an angiotensin converting enzyme inhibitor. Surgery was refused by her parents. We describe here a child with Marfan syndrome in whom significant dilatation of the ascending aorta and severe aortic regurgitation is encountered and major cardiovascular complications of Marfan syndrome were reviewed. PMID- 15529907 TI - Severe cardiovascular features of marfan syndrome in childhood: just another manifestation or a specific entity? PMID- 15529908 TI - Indices of aortic distensibility and coronary flow velocity reserve in patients with different grades of aortic atherosclerosis. AB - OBJECTIVE: The purpose of this study was to evaluate the correlation of the elastic properties of the descending aorta, the coronary flow velocity reserve (CFR) and the grade of aortic atherosclerosis (AA) in patients who have undergone coronary angiography. METHODS: A total of 113 consecutive patients (77 men and 36 women, aged 31-80 years) underwent stress transoesophageal echocardiographic (STEE) assessment of the CFR. The grade of AA and the indices of aortic distensibility were evaluated during the same session of STEE. All patients had chest pain without previous myocardial infarction. Coronary angiography was performed in all cases. RESULTS: The resting systolic and diastolic coronary flow velocities increased, while the coronary flow velocities measured at the peak of stress and the ratio Smax/Srest decreased in parallel with the aortic grade. The CFR and mean CFR were impaired in patients who exhibited aortic intimal thickening, but no further decrease was found in the event of aortic plaque. The elastic moduli E(p) and E(s) increased in parallel with the grade of AA. The occurance of a negative coronary angiogram was more common in patients without AA, but significant left anterior descending coronary artery disease or multivessel disease were more frequent at a higher grade of AA (in cases with aortic plaque). CONCLUSIONS: The CFR was decreased in patients with aortic intimal thickening, but no further decrease was observed in the presence of aortic plaque. The indices of aortic distensibility increased continuously in parallel with the aortic grade. PMID- 15529909 TI - Incidence of stent under-deployment as a cause of in-stent restenosis in long stents. AB - Although stent under-deployment (SU) has been associated with increased risk of in-stent restenosis, little data have been reported on the incidence of SU in patients presenting with clinical in-stent restenosis. In 59 patients referred for vascular brachytherapy and showing angiographic in-stent restenosis, we sought (1) to determine the incidence of SU using standard intravascular ultrasound (IVUS) criteria (2) to evaluate the effects of repeat angioplasty on further stent expansion. Stented length was 32+/-17 mm and diameter stenosis was 75+/-14%. Before re-intervention, the incidence of reduced absolute values of minimal stent cross-sectional area (MSCSA) varied from 69% (< or =8 mm2) to 15% (< or =5 mm2). After re-intervention, the incidence decreased to 24% (< or =8 mm2) (p = 0.0001) and 0% (< or =5 mm2) (p = 0.005). Before re-intervention, SU as assessed by relative criteria varied from 21% (80% mean reference lumen area or 90% minimum distal reference lumen area) to 28% (100% minimum reference lumen area). After re-intervention, the incidence of SU varied from 7% (90% minimum distal reference lumen area) (p = 0.0001 vs. pre) to 24% (55% mean reference EEM area) (p = ns). No change in strut apposition (97% pre vs. 100% post) nor in symmetry index (100% pre vs. post) was noted. From all criteria, the 90 and 100% minimum reference lumen area criteria were the most altered by repeat balloon dilatation, 21% pre vs. 7% post and 28% pre vs. 11% post, respectively. In conclusion, among patients presenting with severe angiographic in-stent restenosis, a significant number showed signs of SU whose incidence varied according to applied criteria. Significant stent re-expansion can be obtained following IVUS-guided repeat angioplasty irrespective of initial SU criteria. PMID- 15529910 TI - Radiation peak skin dose to risk stratify electrophysiological procedures for deterministic skin damage. AB - Ionising radiation is has the potential to cause harm both by increasing the probability future malignancy (stochastic mechanisms) and by direct physical injury (deterministic mechanisms). Several measures have been developed to quantify radiation exposure during a procedure and cardiologists usually refer to fluoroscopic screening time (FST). FST, however, has limitations for predicting deterministic injury which is directly dependant on peak skin dose (PSD). We compared FST to PSD for a range of interventional cardiac electrophysiology procedures. METHODS: All patients undergoing electrophysiology procedures during a 2-month period in our institution were studied. Demographic details, nature of procedure, FST and PSD were measured. The FST to PSD ratio was calculated and compared between patient and procedural factors. RESULTS: 67 procedures on patients (23 female) with body mass index (BMI) of 28 (SD 5) Kg/m2 were studied. Screening times ranged from 0.2 to 96.6 min (median 11.2). PSD ranged from <0.1 to 1108 mGy (median 141). There was a positive correlation between PSD to FST ratio and BMI (r = 0.59, p < 0.001). The PSD to FST ratio was higher in cardiac resynchronization therapy (CRT) devices than single or dual chamber ICDs (p = 0.002). CONCLUSION: FST is not a reliable predictor of deterministic skin injury and in high-risk procedures such as CRT devices and those on individuals of high BMI PSD should be measured. PMID- 15529911 TI - Novel insights into LV remodeling after murine myocardial infarction by in vivo magnetic resonance tissue velocity mapping. PMID- 15529912 TI - Primary percutaneous coronary intervention of anomalous origin of right coronary artery above the left sinus of Valsalva in a case with acute myocardial infarction. Coronary anomalies and myocardial infarction. AB - Anomalous origin of the right coronary artery from the ascending aorta above the left sinus of Valsalva is exceedingly rare. We presented a case with anomalous origin of the RCA above the left sinus of Valsalva with inferior wall myocardial infarction and successful primary percutaneous coronary intervention to this artery which is the first report in the literature. PMID- 15529914 TI - Three-dimensional analysis of in vivo coronary stent--coronary artery interactions. AB - Stent implantation results in important three-dimensional (3D) changes in arterial geometry which may be associated with adverse events. Previous attempts to quantify these 3D changes have been limited by two-dimensional techniques. Using a 3D reconstruction technique, vessel curvatures at end-diastole (ED) and end-systole (ES) were measured before and after stent placement of 100 stents (3 stent cell designs, 6 stent types). After stenting, the mean curvature at ED and ES decreased by 22 and 21%, respectively, and represents a straightening effect on the treated vessel. This effect was proportional to the amount of baseline curvature as high vessel curvature predicted more profound vessel straightening. When analyzed by stent cell design, closed-cell stents resulted in more vessel straightening than other designs (open cell or modified slotted tubes). Stent implantation resulted in the transmission of shape changes to stent ends and generated hinge points or buckling. Stent implantation creates 3D changes in arterial geometry which can be quantified using a 3D reconstruction technique. PMID- 15529913 TI - Aortoiliac angiography during coronary artery angiography detects significant occult aortoiliac and renal artery atherosclerosis in patients with coronary atherosclerosis. AB - PURPOSE: Peripheral atherosclerosis (renal and aortoiliac localizations) are frequently detected in aged patients with concomitant coronary artery disease (CAD): the risk of finding peripheral disease is increased in patients with CAD. Angiography of the aortoiliac vessels performed at the time of coronary angiography may detect any occult renal artery stenosis and atherosclerotic involvement of the aortoiliac segment. We sought to determine utility of performing angiography of aortoiliac and renal arteries during coronary arteriography in patients with known or suspect coronary atherosclerosis. METHODS: Medical records of all patients undergoing combined coronary and aortoiliac angiography between May 1998 and December 2002 was retrospectively reviewed. Moderate to severe arterial stenosis (>50% stenosis), vessel occlusion, aneurismal vessels were noted as significant angiographic findings. Contrast induced nephropathy was defined as a rise in serum creatinine of > or =25% form baseline. RESULTS: In the study period 112 patients (81 males, mean age 68.4+/ 7.8 years) with known or suspected CAD underwent combined cardiac catheterization and aortoiliac angiography. Pretreatment with 0.45% saline at a rate of 1 ml/kg/h for 12 h was administered to all patients. Significant findings were reported in 37 (33%) patients including 14 renal artery stenoses, 8 aortic aneurismal disease, and 15 aortoiliac lesions. Most patients with significant findings had 2 and 3-vessel CAD. A strong correlation was found between the number of risk factors and the frequency of angiographic findings (r = 0.92). Complications include six contrast-induced nephropathy: no case required renal replacement therapy. CONCLUSIONS: Aortoiliac and renal atherosclerosis is frequently associated to multivessel CAD. In selected patients undergoing cardiac catheterization aortoiliac angiography may be practical in detecting occult renal or aortoiliac artery lesions. Further clinical outcome studies are strongly required to support this strategy. PMID- 15529915 TI - The acute administration of trimetazidine modified myocardial perfusion and left ventricular function in 31 patients with ischaemic ventricular dysfunction. AB - Trimetazidine (TMZ) increases the mithocondrial oxidative metabolism and improves Tc-99m sestamibi uptake in myocardial single photon emission tomography (SPECT). The aim of this study was to evaluate whether the acute administration of TMZ improved myocardial perfusion and modified left ventricular ejection fraction (LVEF) in ischaemic left ventricular impairment. METHODS: Thirty-one patients (23 males, age 66 years) with prior myocardial infarction (>6 months) and echocardiographic LVEF < or = 45% underwent coronary angiography, rest basal myocardial SPECT (after 3-day placebo administration) and rest TMZ myocardial SPECT [after 3-day TMZ administration (60mg/die)]. The left ventricle was analysed in 16 segments. The summed placebo score (SPS) and the summed TMZ score (STS) were calculated with a 5-point scale (from 0 = normal uptake to 4 = absent uptake) by two blinded operators. The GATED Tc-99m SPECT was always provided. RESULTS: After TMZ administration GATED LVEF improved from 26.5+/-9.7% to 29.1+/ 11.3% (p = 0.04) and left ventricular end-systolic volume (LVESV) was reduced from 90.2+/-40.7 to 85.6+/-39.2 ml/mq (p = 0.006). Similarly the addition of TMZ to myocardial SPECT significantly reduced the STS compared to SPS (21.5+/-11 vs. 26.6+/-10.5 p = 0.0001). Eleven patients (35.5%) had an echocardiographic LVEF < or = 30%; in these patients who had severe ventricular dysfunction, GATED LVEF and LVESV did not change after TMZ (20.2+/-5.7% vs. 21+/-6.9% p =0.6; 116.7+/ 35.3 ml vs. 112.6+/-32.3 ml p = 0.08, respectively). CONCLUSION: In comparison with placebo, the addition of TMZ to myocardial Tc-99m tetrofosmin SPECT improved myocardial perfusion and LVEF, reducing LVESV. These effects were lost in patients with more severe ventricular dysfunction. PMID- 15529916 TI - A reverse perfusion pattern during Technetium-99m stress myocardial perfusion imaging does not predict flow limiting coronary artery disease. AB - BACKGROUND: A reverse redistribution pattern during myocardial perfusion imaging is most widely described using thallium (Tl-201), when stress images exhibit greater perfusion than rest. Technetium (Tc-99 m) radiopharmaceuticals may also yield a reverse perfusion (RP) pattern, but its significance is uncertain. This study tested the hypothesis that RP correlates with the presence and location of flow limiting coronary stenosis(es). METHOD: We reviewed 842 consecutive Tc-99 m tetrofosmin SPECT stress studies performed at a cardiothoracic centre over a 15 month period. 69 (8.2%) demonstrated RP. Thirty-three patients (age 32-79 mean 56, 17 female) had undergone cardiac catheterisation within 12 months of the scan. Correlation was sought between the presence and location of angiographic stenoses and RP pattern. RESULTS: 10/33 (30.3%) had significant (>60%) coronary stenosis(es); 5 single-vessel, 2 two-vessel and 3 three-vessel disease (3VD). Stenosis location correlated poorly with the RP territory (LAD/Anterior 5/17, RCA/Inferior 1/10, Cx/lateral 0/4 (p = 0.57)). Of the 6 patients with a lesion in the RP territory, 3 had 3VD; 2 of these had a simultaneous reversible defect. All 5 patients with previous myocardial infarction had a simultaneous fixed defect. However only 3/12 with co-existent reversible defects had significant disease. CONCLUSION: The reverse perfusion pattern is a poor predictor of flow limiting coronary disease, and does not correlate with stenosis location in those with significant lesions. Such patients should not undergo invasive investigation purely on the basis of this result. PMID- 15529917 TI - Optimization of ex vivo CT- and MR- imaging of atherosclerotic vessel wall changes. AB - OBJECTIVE: To optimize a methodology for ex vivo imaging of atherosclerotic vessel wall changes using multidetector-row computed tomography (MDCT) and multi contrast magnetic resonance imaging (MRI). METHODS: In phantom studies and studies on intact ex vivo porcine and human hearts, various filling mixtures of MDCT and MRI contrast agents have been evaluated, to enable filling and distension of the coronary arteries for optimal visualization of atherosclerotic vessel wall changes with both techniques. Various proportions of methyl cellulose, iodine-containing CT contrast agent and paramagnetic MR contrast agent containing iron-oxide particles have been tested. Imaging parameters have been optimized for high resolution plaque imaging using a four detector-row CT scanner and a 1.5 T MR system. RESULTS: Phantom studies and studies on ex vivo porcine and human hearts demonstrated optimal proportion of methyl cellulose and CT contrast agent to be 98% vs. 2%, and 75% vs. 25% of methyl cellulose vs. MR contrast agent, respectively. These proportions provided optimal opacification of the vessel lumen in the MDCT images with 250 Hounsfield Units, and good signal suppression within the vessel lumen in the MR images, resembling in vivo imaging techniques. After retrospective matching with histopathology, atherosclerotic lesions of the human ex vivo specimens could be identified on MRI and MDCT images. CONCLUSION: Using an optimized mixture of methyl cellulose, MDCT and MRI contrast agents, visualization of atherosclerotic vessel wall changes is feasible, and applicable to various ex vivo models. PMID- 15529918 TI - Non-invasive, ex vivo imaging of the arterial wall: implications for the understanding of atherosclerotic disease development. PMID- 15529919 TI - Optimization of ex vivo CT- and MR- imaging of atherosclerotic vessel wall changes. PMID- 15529920 TI - Coronary stent imaging with multidetector row computed tomography. AB - Multidetector row Computed Tomography (MDCT) is developing rapidly as a potential non-invasive imaging method for the direct visualization of coronary arteries in patients presenting with suspected coronary artery disease. Recent technical improvements have led to improved spatial and temporal resolution and may even allow evaluation of coronary stents, which has remained difficult due to the small diameter and radio-opaque material. We describe two cases, a 65- and a 72 year old patient. Both patients had previously undergone percutaneous transluminal coronary angioplasty in combination with stent placement and were re admitted to our hospital with recurrent angina. MDCT was performed prior to or after coronary angiography in order to assess stent patency non-invasively. PMID- 15529921 TI - [Incidentally detected renal cancer. Prognostic factors]. AB - OBJECTIVE: To study the clinical and pathological characteristics of incidental renal tumors treated in our center. MATERIAL AND METHODS: A retrospective review is conducted of 318 nephrectomies comparing the clinico-pathological variables of renal tumors diagnosed incidentally with those of symptomatic renal tumors. The factors influencing disease-free survival are analyzed in both groups. RESULTS: In our experience, although incidental renal tumors presented better survival than symptomatic ones owing to their better pathological state and tumor grade, incidental diagnosis was not an independent influencing factor in the multivariate study. Only when patients were studied who did not present metastases on diagnosis did incidental diagnosis become an influencing factor very close to statistical significance. CONCLUSIONS: Incidental diagnosis is not an independent prognostic factor. PMID- 15529922 TI - [Treatment of post radical prostatectomy erectile dysfunction with sildenafil in a non-selected group of patients]. AB - OBJECTIVE: To evaluate erectile disfunction (ED) after radical prostatectomy in a non selected group of patients and their response to sildenafil. METHODS: We included our patients who were operated on between 1998 and 2001. The patients filled in a modification of IIEF (mIIEF) before the RP operation. We tried sildenafil with doses of 100 mg in 3 different periods: 3-6, 12 and 18-24 months after the RP operation. In the event of a complete response they filled in the mIIEF again. RESULTS: The mean age of the patients was 62.8 years old and the mean observation period was 31.7 months. Only 111 (62%) of the total number of patients operated on wanted treatment and only 90 took sildenafil in the proposed protocol; 27 (30%), 18 (20%) and 45 (50%) patients had a complete response, a partial response or no response to the sildenafil respectively. The mIIEF showed 6.5+/-5.7 points less than the mIIEF pre-treatment. In the univariate analysis, the preservation of bundles, the presence of a previous partial response and the presence of the previous tumescence were significantly related to the complete response; however, only tumescence kept its value in the multivariate study. CONCLUSIONS: Urologists must involve themselves in the early rehabilitation of the erectile function even in those patients where preservation of the NVB was not possible. One must always try to preserve, uni or bilaterally, whenever the patient desires preservation of EF. The response to sildenafil is better after the first year and in patients who have previous tumescence. PMID- 15529923 TI - [Treatment of erectile dysfunction with penile prothesis. Our experience]. AB - INTRODUCTION AND OBJECTIVES: The aim of this study was to review the current situation of penile prosthesis in the treatment of erectile dysfunction and to analyze the indications, degree of acceptability and complications in our experience. MATERIAL AND METHODS: From 1984 to 2003, 48 implants were inserted in men suffering from impotence due to different etiologies. The mean age was 55 years. Vascular disease, Diabetes and pelvic surgery were the most common pathologies recorded. The type of prothesis inserted were malleable inactive in 7 cases (14.58%), Jonas6, Acu-form1. Inflable active with 3 components in 19 cases (39.58%), AMS 700 plus17, Alpha2. Inflable active integrated in 5 cases (10.41%) Hydroflex4, Dynaflex1. Inflable active with 2 components in 17 cases (35.4%), Ambicor2, Mark II15. RESULTS: Infection of the prosthesis was observed in 6.25% of the cases. This infectious complication required removal of the prosthesis. 4.16% of the cases complained of mechanical failure that required change of the prosthesis. 80% of the patients were satisfied with the results. CONCLUSIONS: Penile prostheses continue to be an effective and valid therapeutic alternative in impotent. Although the complication rate is low, adequate information must be provided to the patient. PMID- 15529924 TI - [Quality analysis of the statistical used resources (material and methods section) in thesis projects of a university department]. AB - INTRODUCTION AND OBJECTIVES: Studies about quality in thesis and investigation projects in biomedical sciences are unusual, but very important in university teaching because is necessary to improve the quality elaboration of the thesis. The objectives the study were to determine the project's quality of thesis in our department, according to the fulfillment of the scientific methodology and to establish, if it exists, a relation between the global quality of the project and the statistical used resources. MATERIAL AND METHODS: Descriptive study of 273 thesis projects performed between 1995-2002 in surgery department of the Zaragoza University. The review realized for 15 observers that they analyzed 28 indicators of every project. Giving a value to each of the indicators, the projects qualified in a scale from 1 to 10 according to the quality in the fulfillment of the scientific methodology. RESULTS: The mean of the project's quality was 5.53 (D.E: 1.77). In 13.9% the thesis projects was concluded with the reading of the work. The three indicators of statistical used resources had a significant difference with the value of the quality projects. DISCUSSION: The quality of the statistical resources is very important when a project of thesis wants to be realized by good methodology, because it assures to come to certain conclusions. In our study we have thought that more of the third part of the variability in the quality of the project of thesis explains for three statistical above mentioned articles. PMID- 15529925 TI - [Antimicrobial susceptibility of Escherichia coli isolated from bacteriurias in Bierzo health area during 2003]. AB - OBJECTIVE: Sensitivity to antibiotics of Escherichia coli isolated from intrahospital and extrahospital bacteriurias is analyzed. MATERIAL AND METHODS: Sensitivity to nineteen antibiotics used in the clinical practice was determined in strains of Escherichia coli isolated from intrahospital and extrahospital bacteriurias. RESULTS: The prevalence of in-vitro susceptibilities to antibiotics were (hospital, state health service clinic, health center-primary-care): imipenem (100%-100%-100%; p=NS), amikacin (100%-100%-99.7%; p=NS), fosfomicyn (98.6%-98.4%-99.6%; p=NS), cefepime (96%-96.9%-98.3%; p=NS), piperacillin tazobactam (96%-95.3%-96.6%; p=NS), aztreonam (93.5%-94.7%-97.7%; p<0.001), ceftazidime (93.5%-94.3%-97.8%; p<0.001), cefotaxime (93.1%-95%-97.7%; p<0.001), cefixime (92.7%-94.6%-96.7%; p<0.05), nitrofurantoin (92%-94.7%-94.7%; p=NS), cefuroxime (88.4%-93.1%-95%; p<0.001), amoxicillin-clavulanic (87.7%-88.7%-93.8%; p<0.001), tobramicyn (87%-93.7%-93.8%; p<0.001), gentamcin (85.9%-92.8%-93%; p<0.001), cefazolin (84.4%-88.7%-91.6%; p<0.01), ciprofloxacin (63.8%-71.4% 78.4%; p<0.001), norfloxacin (63%-70.8%-78.2%; p<0.001), cotrimoxazole (65.2% 68.6%-74.9%; p<0.01) and ampicillin (35.5%-42.5%-47.8%; p<0.01). (*NS= No significant differences). CONCLUSIONS: The knowledge of the sensitivity of uropathogens to antimicrobians can allow us to initiate empirical therapy of urinary tract infections. PMID- 15529926 TI - [Study of the angiogenesis as prognostic factor of pT1G3 bladder tumours]. AB - INTRODUCTION AND OBJECTIVES: Angiogenic activity has been considered like prognostic factor in several solid tumors. This activity can be analysed by two ways: immunohistochemical determination of molecules that activate/inhibit angiogenesis or quantitive measure of microvascular density (MD). Our objective is to determine the prognostic value of Vascular Endothelial Growth Factor (VEGF) and Microvascular Density (MD) in pT1G3 bladder tumours. MATERIAL AND METHODS: We have studied retrospectively 83 patients with pT1G3 tumors treated by TUR + endovesical instillations with a follow up of 3 years at least. We analysed VEGF expression monoclonal antibody No. 360P. To determine MD we have marked vessels with FVIII antibody and detected "hot spots" areas. The number of microvessels is quantited by a digital image analyser excluding those that have more than 50 micras of diameter. We established the correlation of these findings with recurrence, progression and survival by using Chi-square test and Kaplan-Meier curves (log-rank). RESULTS: Average follow up was 58 +/- 28 months. We have established like cut-off 50% of tumor cells (VEGF) and 30 microvessels/fields (MD). Chi-square test did not show correlation with survival neither recurrence but it was positive for progression p(VEGF) 0.048 and p(DM) 0.021. Kaplan Meier curves determined significative differences only for free of progression time respect to MD (p 0.038). CONCLUSIONS: We did not find statistically significant value for recurrence nor survival. Just MD reached prognostic value for progression. More studies and multivariant analysis are required to determine the clinical utility of MD, specially in order to make more aggressive therapeutic options in this kind of patients. PMID- 15529927 TI - [Cystic testicular lesions in infancy]. AB - The present article reports a case 11 month-old infant with a right intratesticular cyst. We analyze the etiology, differential diagnosis and management off all cystic lesions of the pediatric testis. Patient age at presentation, examination features, tumor markers and sonographic appearance may assist in making a presumptive and occasionally definitive diagnosis preoperatively. The differential diagnosis include intratesticular simple cyst, epidermoid cyst, tunica albuginea cyst, testicular teratoma, juvenil granulosa cell tumor-gonadal stromal tumor, cystic dysplasia of the rete testis, cystic lymphangioma, and testicular torsion. Usually enucleation is the best treatment. A thorough understanding of potentially cystic testis lesions in children leads to the best management choices and often to preservation of a substantial portion of the affected testis. PMID- 15529928 TI - [Pneumoscrotum secondary to a massive subcutaneous emphysema after expontaneous pneumothorax drainage]. AB - We present a case of a pneumoscrotum and pneumopenis which appeared after a trocar drainage placement for treatment of a recidivated expontaneous pneumothorax. In spite of is an entity with easy diagnosis and conservative management, maybe be confused with other more aggressive entities which treatment should be invasive. PMID- 15529929 TI - [Complex renal cyst. A new case]. AB - Approximately 15% of cases of renal cell carcinoma present cystic configuration on radiologic and pathologic examination. These lesions are often difficult to differentiate from the multiloculated renal cyst or other benign cystic lesions such as hemorrhagic cyst and so on. We report a case of multilocular cyst of the right kidney complicated with clear cell type renal cell carcinoma in which MRI suggested benign cyst. The diagnosis of complicated benign cyst must be proposed very cautiously and the persistence of doubtful images justifies surgical exploration. PMID- 15529930 TI - [Melanosis of the bladder: a very uncommon entity]. AB - Melanosis of the bladder is a rare entity characterized by anomalous deposits of melanin-containing pigment on the vesical urothelium without melanocytic atypia or associated malignant melanoma. To our knowledge and following such strict criteria, there are only five cases previously reported. The biological potential of this entity is uncertain because of the rarity of their. We report one case of simple melanosis of the bladder confirmed by histological examination and inmunohistochemistry analysis. We believe that recognition of this entity is important. PMID- 15529931 TI - [Urinary bladder xanthoma. A cystoscopic finding]. AB - Urinary bladder xanthoma (UBX) is an infrequent lesion that has been very rarely referred to in medical literature. We describe a case of UBX that was associated with a low-grade transitional cell carcinoma of the urinary bladder. The cystoscopic aspect of this lesion was so characteristic that diagnosis could be suggested at cystoscopy, while histological examination confirmed this. PMID- 15529933 TI - [Retroperitoneal malignant fibrous histiocytoma]. AB - We report a new case of retroperitoneal tumor: a malignant fibrous histiocytoma, in its less aggressive histologic type, the mixoid variety. This is a recently isolated histologic presentation from other sarcomas, with a low incidence, non specific clinic findings and poor prognosis due to its tendency to reappear and to produce metastasis. PMID- 15529932 TI - [Laparoscopic pyelolithotomy in a pelvic kidney]. AB - INTRODUCTION: Pelvic renal ectopia bearing stones is a particularly rare case of complex calculi. Several percutaneous and laparoscopic procedures had been proposed in the last years to treat this challenge problem in a minimally invasive way. CLINICAL CASE: We describe the procedure of laparoscopic pyelolithotomy under general anesthesia, using three port sites, in a male patient with symptomatic stones in a left pelvic kidney with normal function. Two stones were successfully removed, with the help of flexible nephroscopy through a port, and the pyelotomy was closed with intracorporeal suture. COMMENT: Laparoscopic pyelolithotomy of a pelvic kidney is a minimally invasive technique that is not difficult to perform, making laparotomy unnecessary. Laparoscopic technique must be take into account when dealing with the infrequent complex stone patient candidate to an open stone surgery. A review of the Spanish bibliography shows this to be the second reported case of pelvic kidney calculi treated by laparoscopic pyelolithotomy. PMID- 15529934 TI - [Retroperitoneal neurilemmona]. AB - Schwannoma is an uncommon tumor arising from neural sheath of peripheral nerves. Histological findings showing Antoni A and B zones are the main diagnostic features. We report a new case of a retroperitoneal located one, which is not its most usual place to appear, in a 60 year old male, presenting left flank pain. He received surgical treatment, having a satisfactory evolution. PMID- 15529936 TI - [Skin burn caused by shock-wave lithotripsy]. PMID- 15529939 TI - [Budd-Chiari syndrome]. PMID- 15529940 TI - The hungry bone: expected and unexpected. PMID- 15529941 TI - [Iron overload disease: recent findings]. AB - Iron overload diseases are due to a progressive increase in total body iron stores that leads to deposition of iron in parenchymal organs and to subsequent damage to these organs. The commonest inherited form of iron overload is hereditary hemochromatosis (HH), an autosomal recessive disorder affecting the white population. Although in the western world and in northern Europe the majority of cases of HH are associated with an HFE gene mutation (C282Y and H63D), there are families with a familial iron overload disorder in whom neither the C282Y nor the H63D mutations were found. Recently, other forms of HH that are not related to HFE, but are due to mutations in genes coding iron transport proteins (ferroportin-1, TfR2, hepcidin) have been described. The clinical presentation of the disorder is highly variable, depending on the severity of iron overload. In fact, the inappropriate absorption and deposition of dietary iron may result in the development of hepatic and non-hepatic end-organ injury, leading to liver cirrhosis, hepatocellular carcinoma, diabetes, arthritis, skin pigmentation and cardiac diseases. HH and its sequelae are preventable with an early diagnosis and treatment. Patients with evidence of iron overload, a family history of HH or other risk factors should be screened by genotype testing for the HFE mutation. Nowadays, HH is recognized as being a complex genetic disease with probable significant environmental and genetic modifying factors, such as hepatitis C virus infection and alcohol abuse, and it has been shown that HFE mutations represent an independent risk factor for fibrosis and cirrhosis in chronic hepatitis C. PMID- 15529942 TI - [Improving compliance in internal medicine: the motivational discussion]. AB - Compliance to treatment is of crucial importance in medicine. High levels of noncompliance to treatment have been reported in the most relevant diseases such as hypertension, coronary artery disease, diabetes and asthma. The aim of this article was to fully evaluate the lack of adherence to treatments in internal medicine. The prevalence of noncompliance and methods to cope with it are presented. The theory of the stages of change is thoroughly examined. Motivational interviewing in low-compliant patients is illustrated dealing both with the theoretical principles and practical applications. Several communication blocks which interfere with the relationship between doctor and patient are finally presented through concrete examples. PMID- 15529943 TI - [Arterial hypertension and cognitive deficit]. AB - Cognitive impairment and dementia are more and more common in the elderly. The first begins, it advances silently and it leads to dementia in few years. Arterial hypertension represents the most important cerebrovascular risk factor after age. In numerous studies an inverse relationship between blood pressure values and cognitive performance emerges: it is possible that arterial hypertension plays a role in the pathogenesis of cognitive decline. Even in asymptomatic subjects the magnetic resonance signs of cerebral damage accompany cognitive impairment development. Antihypertensive therapy influence on cognitive function represents a subject of actual interest. The most studied drugs are calcium antagonists and ACE-inhibitors; they seem to have a protective effect on cognitive impairment, with regard to diuretics and beta-blockers. It would be important to study hypertensive patients, above all young asymptomatic hypertensives, even about cognitive functions, to prevent and consider cognitive decline and effective organ damage. PMID- 15529944 TI - Immunoglobulin gene rearrangements and in vitro growth of stromal cells are prognostic indexes in B-chronic lymphocytic leukemia. AB - B-chronic lymphocytic leukemia is a disease characterized by an accumulation of monoclonal B cells that are resistant to apoptosis. In chronic lymphocytic leukemia, the prognosis depends on the stage of the disease, according to the classifications of Rai and Binet. However, in recent years, the number of patients with very early disease (stage 0 of Rai) and without any clinical symptom, has considerably increased because of the extensive use of automatic apparatus for leukocyte counting and immunophenotypic analysis of lymphocytes. It has become, therefore, useful to find new prognostic criteria particularly for these patients. In the present study, 30 patients with B-chronic lymphocytic leukemia were investigated for stage of the disease, survival, immunoglobulin gene rearrangements, presence of nurse like cells in in vitro cultures and spontaneous clinical lymph node regression. We observed that all these criteria are useful prognostic indexes for the disease. PMID- 15529945 TI - [Bone mass, bone metabolism, and prevalence of spinal fractures in recipients of allogenic bone marrow transplantation for leukemia]. AB - The aim of this study was to evaluate in 65 patients, who had previously undergone allogenic bone marrow transplantation (ABMT), the bone mineral density (BMD), the skeletal turnover and the prevalence of vertebral fractures. At the moment of recruiting, 10 of 65 transplanted subjects (15.3%) presented with signs of rejection of the transplanted tissue, thus they were excluded. The remaining 55 patients (21 males, 34 females, mean age 30.8 +/- 6.4 years), with a follow-up of 60 +/- 9 months after the transplant and without any treatment inducing osteopenia, underwent ABMT respectively for: chronic myeloid leukemia (n = 24); acute myeloid leukemia (n = 18); acute lymphatic leukemia (n = 13). One hundred and ten healthy control subjects (42 males and 68 females, mean age 31.0 +/- 3.7 years) matching with the patients for age, weight and height, were successively enrolled. All the participants were submitted to a densitometric evaluation (DEXA) of lumbar spine (L1-L4), of femoral neck and total femur; besides some skeletal metabolism parameters were dosed, such as: total alkaline phosphatase, bone alkaline phosphatase and urinary excretion of C-terminal telopeptide fragments normalized to creatinine. On the contrary, the morphometric evaluation, performed through a lateral dorsolumbar radiography, was actually carried out only in patients who had already undergone ABMT. The L1-L4 BMD study showed: 10/55 osteoporotic (18.1%), 19/55 osteopenic (34.5%) and 26/55 normal patients (47.4%). In transplanted patients BMD values, obtained at the three considered sites, resulted significantly reduced (p < 0.01) in comparison to controls. Moreover, in patients who underwent ABMT, a statistically significant increase was observed, in comparison to healthy subjects, in total alkaline phosphatase (p < 0.01), in bone alkaline phosphatase (p < 0.01) and in urinary excretion of C terminal telopeptide fragments normalized to creatinine levels (p < 0.001). Seven of the 55 transplanted patients (12.7%) presented at the moment of Rx morphometric evaluation at least one vertebral fracture: 6 of whom were affected by osteoporosis and 1 by osteopenia. In conclusion, the subjects who had previously undergone ABMT maintain, even at a certain time after the transplant and without any rejection, an increased skeletal turnover and BMD values meanly lower than normal, leading to an increased risk for vertebral fracture. PMID- 15529946 TI - [Budd-Chiari syndrome with fatal outcome in a patient with polycythemia vera and antiphospholipid antibody syndrome]. AB - We report the case of a 41-year-old woman, affected by Vaquez syndrome, admitted to our hospital for a severe pain in the right hypochondrium, suddenly followed by hepatomegaly and ascites. The clinical and laboratory data were suggestive of hepatic insufficiency and abdominal ultrasonography, integrated by color Doppler and computed tomography, revealed an interrupted hepatic venous outflow. In addition a spontaneous prolonged partial thromboplastin time was present and the patient was found to be positive for lupus anticoagulant. A transient clinical improvement, with a partial reperfusion of suprahepatic veins, was achieved with medical treatment by using anticoagulants, diuretics and paracentesis. However, the patient showed a subsequence of suprahepatic venous thrombosis, although two transjugular intrahepatic portosystemic shunts with stent placement and local thrombolysis were performed. The polycythemia vera is a disease mainly associated with Budd-Chiari syndrome but, in our patient, the thrombotic event occurred in spite of normal values of hematocrit and platelet count. Certainly in this case the lupus anticoagulant positivity represents an additional thrombogenic factor. Nowadays the antiphospholipid antibody syndrome is a recognized and not unusual cause of Budd-Chiari syndrome but, to our knowledge, this is the first case characterized by the presence of polycythemia vera and antiphospholipid antibody syndrome to be reported. PMID- 15529947 TI - Kinetics of the circulating levels of bone alkaline phosphatase in a case of hungry bone disease following total parathyroidectomy. AB - We present the case of a 55-year-old patient who underwent total parathyroidectomy for severe hyperparathyroidism unresponsive to medical therapy, 4 years after having started hemodialysis treatment. It was decided to perform total parathyroidectomy because at macroscopic evaluation the parathyroid glands appeared completely compromised. After surgery, the patient developed hungry bone disease, characterized by severe hypocalcemia and hypophosphatemia. After parathyroidectomy, serial measurements were made for the long-term monitoring of the calcemia, phosphatemia and the serum levels of intact parathormone and bone alkaline phosphatase, a marker of bone turnover that mainly expresses bone formation. There was initially a slight decrease in the circulating levels of bone alkaline phosphatase as the calcemia dropped dramatically, then a new increase that anticipated the subsequent calcemia increase and finally, 6 months later, a decrease to very low values. We believe that the calcemia and blood bone alkaline phosphatase could be useful for the laboratory monitoring of the hungry bone state, providing information which may be useful to avoid excessive calcium administration and the dangerous consequences such as soft-tissue calcification. PMID- 15529948 TI - Alterations of the sympathovagal balance evaluated by heart rate variability in a rare case of adult Still's disease. AB - We present a rare case of adult Still's disease with cardiac involvement (myocarditis and coronary arteritis). The autonomic nervous system function was evaluated by heart rate variability (HRV) analysis performed by 24-hour electrocardiographic recording during the acute phase of the disease and the remission (after 1 month and 1 year). The HRV parameters were studied in the time (standard deviation of all NN intervals-total power - SDNN, square root of the mean of the sum of the squares of the differences between adjacent NN - RMS-SD and HRV index) and frequency domains (low frequency, high frequency, and low frequency/high frequency). The results of the analysis of the HRV highlight that in the acute phase of the disease with cardiac involvement the autonomic nervous system is globally altered, with modifications of the sympathovagal balance, due to impairment of the parasympathetic component. This trend tends to persist in the short period (1 month), but seems to resolve completely within 1 year. An altered sympathovagal balance should be considered as a possible marker of vasculitis-related ischemia. PMID- 15529949 TI - [Scientific medicine and alternative medicine: the problem of the separation line]. PMID- 15529950 TI - [The health world's uneasiness: crisis of medicine or crisis of philosophy?]. PMID- 15529951 TI - [Fever of unknown origin and monoclonal gammapathy with unclear significance. Report of 2 cases with different pathology and clinical course]. PMID- 15529952 TI - Vessel-based non-rigid registration of MR/CT and 3D ultrasound for navigation in liver surgery. AB - OBJECTIVE: Computer assisted planning of liver surgery based on preoperative computed tomography (CT) or magnetic resonance imaging (MRI) data can be an important aid to operability decisions and visualization of individual patients' 3D anatomy. A navigation system based on intraoperative 3D ultrasound may help the surgeon to precisely localize vessels, vascular territories or tumors. The preoperative planning must be transferred to the intraoperative ultrasound data and thus to the patient on the operating table. Due to deformations of the liver between planning and surgery, a fast non-rigid registration method is needed. MATERIALS AND METHODS: A feature-based non-rigid registration approach based on the centerlines of the portal veins has been developed. The combination of an iterative closest point (ICP) approach and Multilevel B-Spline transformations offers a fast registration method. The vessels are segmented and their centerlines extracted from preoperative CT/MRI and intraoperative 3D Powerdoppler ultrasound data. Anatomical corresponding points on the centerlines of both modalities are determined in each iteration of the ICP algorithm. The search for corresponding points is restricted to a given search radius and the direction of the vessels is incorporated. RESULTS: The algorithm has been evaluated on two transcutaneous and one intraoperative clinical ultrasound data set from three different patients. Only a very few vessel segments were not assigned correctly compared to manual assignments. Using non-rigid transformations improved the root mean square target registration error of the vessels by approximately 3-5 mm. CONCLUSIONS: The proposed registration method is fast enough for clinical application in liver surgery. Initial accuracy results are promising and must be further evaluated, particularly in the operating room. PMID- 15529953 TI - Fast and adaptive finite element approach for modeling brain shift. AB - OBJECTIVE: In this paper we introduce a finite element-based strategy for simulation of brain deformation occurring during neurosurgery. The phenomenon, known as brain shift, causes a decrease in the accuracy of neuronavigation systems that rely on preoperatively acquired data. This can be compensated for with a computational model of the brain deformation process. By applying model calculations to preoperative images, an update within the operating room can be performed. METHODS: One of the crucial concerns in the context of developing a physical-based model is the choice of governing equations describing the physics of the phenomenon. In this work, deformation of brain tissue is expressed in terms of a 3D consolidation model for a linearly elastic and porous fluid. The next crucial issue is ensuring stable calculations within the chosen model. For this purpose, we developed a special technique for generating the underlying geometry for the simulation. With this technique an unstructured grid consisting of regular tetrahedra is created, whereupon time-dependent finite element simulation is performed in an adaptive manner. RESULTS: We applied our algorithm to preoperative MR scans and investigated the value of the method. Due to the adaptivity of the method, only 5-10% of the computing time was needed as compared to traditional finite element approaches based on a uniformly subdivided grid. The results of the experiments were compared to the corresponding intraoperative MR scans. A close match between the computed deformation of the brain and the displacement resulting from the intraoperative data was observed. CONCLUSION: A model-based approach for the simulation of brain shift is presented. In this computational model the brain tissue is described as an elastic and porous material using Biot consolidation theory. Validating experiments conducted with MR data provided promising results. PMID- 15529954 TI - Development and first patient trial of a surgical robot for complex trajectory milling. AB - OBJECTIVE: Today's surgical robots normally perform "simple" trajectories, e.g., assisting as tool-holding devices in neurosurgery, or milling linear paths for cavities in total hip replacement. From a clinical point of view, it is still a complex undertaking to implement robots in the operating room. Until now, robot systems have not been used in patient trials to mill "complex" trajectories, which involve many positional and orientation changes and are often necessary in cranio-maxillofacial (CMF) surgery. This paper presents the RobaCKa surgical robot system, which allows more precise execution of surgical interventions and milling of "complex" trajectories. MATERIALS AND METHODS: The main components of the RobaCKa system are a (former) CASPAR robot system, a POLARIS system, and a force-torque sensor. RESULTS: In the first patient trial (April 2003) the planned trajectory was executed with an error of 0.66 +/- 0.2 mm. CONCLUSIONS: The use of former industrial robots for surgical applications is possible but complex. The advantages are improved precision and quality and the possibility of documentation. The use of such systems is normally limited to research institutions or large clinics, because it is hardly possible to implement the necessary technical and logistic efforts in routine surgical work. PMID- 15529955 TI - Identification of milling parameters for manual cutting of bicortical bone structures. AB - Within the framework of the development of a new neurosurgical robot system, parameters of conventional manual neurosurgical bone milling had to be determined. These results were used as a reference for the design and validation of the robotic versus manual milling task. Bovine scapulae were used for the tests because their bicortical structure is similar to the bone structure of the skull. The exercise was to cut defined geometries that had been registered on the bone prior to the start of the milling operation. The geometries had to be milled with a depth of 3 mm, which corresponded to the radius of the ball of the cutter. Different parameters like tool position, rotary speed, temperature, forces and time were registered. Eleven experienced neurosurgeons with practical experience of 80-1200 skull operations participated in this study. First results show a large variation in depth along the line. The lateral deviation was up to 5 mm, the depth error up to 2.5 mm, the tool temperature was 22 degrees C to 65 degrees C, and rotary speed varied from 15,000 to 80,000 rpm. Registered forces had maxima of 16 N in the feed direction and 21 N normal to the surface; average forces were approximately 1-2 N. PMID- 15529956 TI - CT-based preoperative analysis of scapula morphology and glenohumeral joint geometry. AB - OBJECTIVE: To investigate the geometry of the glenohumeral joint using three dimensional (3D) models; define landmarks, planes, angles and regions of interest; and analyze the exact morphology of the scapula. MATERIALS AND METHODS: We reconstructed 3D scapula models based on computed tomography (CT) data sets of 12 healthy controls. Three-dimensional models were reconstructed using the 3D Slicer2 (Surgical Planning Lab, Boston, MA), which provides interactive measurement. The 3D model and measuring tools can be freely shifted and rotated in all planes. RESULTS: The average length of the scapulas was 209.58 +/- 26.72 mm (left) and 223.68 +/- 47.74 mm (right); width was 124.03 +/- 13.28 mm (left) and 141.44 +/- 27.78 mm (right); and volume was 121.38 +/- 12.02 ml (left) and 130.24 +/- 20.86 ml (right). The glenoid dimensions were 34.34 +/- 5.62 mm (left) and 36.44 +/- 7.36 mm (right) (anteroposterior); and 49.16 +/- 7.68 mm (left), 51.46 +/- 10.07 mm (right) (superoinferior). For all average values, scapulas from male controls were significantly larger than those from females, and right sides were larger than left sides (p < 0.05). The glenoid version was nearly the same for male/female and left/right (left = 9.02 +/- 3.89 degrees retroversion; right = 8.26 +/- 3.72 degrees retroversion) (p > or = 0.05). CONCLUSION: The 3D measurement of scapula morphology compared with geometry of the glenohumeral joint leads to a more precise planning of shoulder surgery. As new shoulder prostheses are intended to reconstruct the normal anatomy as closely as possible, such exact 3D measurements may be used for optimization. PMID- 15529957 TI - A virtual-reality-based haptic surgical training system. AB - To improve training facilities for surgeons, a surgical training system based on virtual reality techniques has been developed. The goal of the developed system is to improve education of surgeons by making the knowledge of expert surgeons directly available to trainees. The system realizes two different approaches: the library and the driving school paradigm. In its current form, the system consists of two modules. The main module combines the virtual reality kernel KISMET, a visual and haptic display, and a database of different operations and/or techniques. The master station is a copy of the input and output facilities of the main module. Both modules communicate by a TCP/ IP-based connection. Initial tests demonstrated the feasibility of the chosen framework. Further developments include the gathering of data not only from virtual reality but also from real operations. Robotic-assisted surgery provides an attractive way of accomplishing this. PMID- 15529958 TI - [Sarcoidosis (Boeck's disease). Inflammatory granulomatous systemic disease which involves especially the lung and lymph nodes]. PMID- 15529959 TI - [What is your diagnosis? Pseudocyanosis due to amiodarone]. PMID- 15529960 TI - [The red face]. AB - Facial erythema may not only present clinically as a distinct entity, but can also be a symptom of other diseases. It is seen in common dermatoses such as eczema, psoriasis, acne and urticaria, as well as in rarer conditions such as disorders of keratinization, infectious diseases, porphyrias and neoplasia. Facial erythema may also present as a symptom of carcinoid syndrome, drug allergies, cardiac disease or in rare cases as a feature of Bloom's syndrome, sarcoidosis, lymphoma, amyloidosis and other disease processes. We would like to concentrate on the practical aspects of facial erythema as a presenting symptom, rather than discussing every disease in detail. PMID- 15529961 TI - [The red tympanic membrane]. AB - An otoscopic red tympanic membrane is for the examiner a challenge. A short directed history, as well as a few functional examinations associated to a systematic otoscopy allow in most cases to make the otological diagnosis. The differential diagnosis for a red tympanic membrane is very large, including malformations, traumas, infections, and even tumors and other degenerative pathologies. The first diagnosis to think about is an infectious disease. On the other side, there are rare pathologies which, even for experienced examiners are quite seldom. PMID- 15529962 TI - [Unclear weight loss. Basedow's disease resp. Graves' disease]. PMID- 15529963 TI - [Protracted fever in initial suspicion of infection of the upper airways. Acute CMV-mononucleosis in immunocompetent patient]. PMID- 15529964 TI - [VALUE study underscores the significance of reaching aimed blood pressure values for minimizing cardiovascular risks]. PMID- 15529965 TI - Behavioral syndromes: an intergrative overiew. AB - A behavioral syndrome is a suite of correlated behaviors expressed either within a given behavioral context (e.g., correlations between foraging behaviors in different habitats) or across different contexts (e.g., correlations among feeding, antipredator, mating, aggressive, and dispersal behaviors). For example, some individuals (and genotypes) might be generally more aggressive, more active or bold, while others are generally less aggressive, active or bold. This phenomenon has been studied in detail in humans, some primates, laboratory rodents, and some domesticated animals, but has rarely been studied in other organisms, and rarely examined from an evolutionary or ecological perspective. Here, we present an integrative overview on the potential importance of behavioral syndromes in evolution and ecology. A central idea is that behavioral correlations generate tradeoffs; for example, an aggressive genotype might do well in situations where high aggression is favored, but might be inappropriately aggressive in situations where low aggression is favored (and vice versa for a low aggression genotype). Behavioral syndromes can thereby result in maladaptive behavior in some contexts, and potentially maintain individual variation in behavior in a variable environment. We suggest terminology and methods for studying behavioral syndromes, review examples, discuss evolutionary and proximate approaches for understanding behavioral syndromes, note insights from human personality research, and outline some potentially important ecological implications. Overall, we suggest that behavioral syndromes could play a useful role as an integrative bridge between genetics, experience, neuroendocrine mechanisms, evolution, and ecology. PMID- 15529966 TI - Comparison of different attenuation strategies in development of a Salmonella hadar vaccine. AB - The purpose of this work was to develop a live, attenuated vaccine strain to protect chickens against colonization by group C Salmonella. We constructed two candidate vaccines: a deltacya deltacrp derivative and a deltaphoP derivative of Salmonella hadar. White Leghorn chickens were vaccinated at day of age and at 2 wk with one of the two strains. A nonvaccinated group served as a control. At 4 wk of age, all birds were challenged with wild-type S. hadar and necropsied 6 days later. Numbers of S. hadar in the ceca were determined. Enzyme-linked immunosorbent assay-derived serum immunoglobulin G responses against S. hadar lipopolysaccharide indicated that both strains induced a serum antibody response. The average optical density450 for birds vaccinated with the deltaphoP or deltacya deltacrp derivatives was 0.456 and 0.881, respectively. Although the deltacya deltacrp derivative induced higher levels of serum antibody, it did not provide an immune response protective against colonization by S. hadar. Conversely, birds vaccinated with the deltaphoP strain showed significant protection against S. hadar challenge. Seventy percent of the nonvaccinates, 60% of the deltacya deltacrp vaccinates, and 15% of deltaphoP vaccinates were positive for S. hadar in tissues. In a second experiment, birds were vaccinated with either the deltaphoP strain or buffer and challenged with a 10-fold higher dose than in the first experiment. After challenge, all of the birds in both groups were colonized. The geometric mean number of cecal S. hadar isolated from the control group was 1.0 x 10(6) colony-forming units (CFU)/g, and from the vaccinated group, this value was 32 CFU/g, indicating a four to five log reduction in colonization by the challenge strain. PMID- 15529967 TI - Differentiation of avian poxvirus strains on the basis of nucleotide sequences of 4b gene fragment. AB - Investigations for detection and differentiation of nine avian poxviruses (APVs) were carried out by the use of a polymerase chain reaction (PCR) combined with restriction enzyme analysis (REA) and further nucleotide sequence analysis. With one primer set, which framed a region within the fowl poxvirus 4b core protein gene, we were able to detect APV-specific DNA from 19 tested strains and isolates belonging to five defined Avipoxvirus species and four previously undefined isolated species. PCR results revealed no recognizable differences in size of amplified fragments among the different APVs. REA of PCR products with MseI and EcoRV allowed us to differentiate most of the tested avipox species. Nucleotide sequence analysis of the amplified fragments showed a nucleotide similarity of 72%-100% among the different species. Phylogenetic analysis documented five distinguishable sequence clusters in accordance with results obtained by REA. PCR in combination with REA and sequencing of the amplified fragments is a rapid and effective diagnostic system, and it is a new approach to refine epidemiologic studies of APV infections. PMID- 15529968 TI - Pasteurella multocida infection in heterophil-depleted chickens. AB - The present study was aimed at elucidating the role of heterophil granulocytes during the initial infection with Pasteurella multocida subsp. multocida in chickens. Chickens (17 and 19 wk old) were depleted of their heterophil granulocytes by 5-fluorouracil treatment. When the heterophil blood counts were significantly reduced, the birds were inoculated intratracheally with 1.8-4.3 x 10(4) colony-forming units of P. multocida. Twelve, 24, or 48 hr postinoculation, the birds were euthanatized and examined for macroscopic and histologic lesions in the lungs. Bacterial invasion was determined by culture of P. multocida from the spleen. Recruitment of heterophils into the respiratory tract during infection was found to contribute considerably to the lung lesions in chickens and was found to mediate tissue damage, possibly allowing a more rapid systemic spread of P. multocida. However, during progression of the infection, the heterophil-mediated necrosis in chickens seemed to stimulate giant cell demarcation of infected lung tissue, which coincided with the clearance of P. multocida from the spleen, thus hampering further invasion. Consequently, heterophil activation plays a dual role for the outcome of a P. multocida infection in chickens, where it initially seems to promote invasion and systemic spread but subsequently helps limit the infection by giant cell formation and bacterial clearance. PMID- 15529970 TI - Morphologic changes in chicken cells after in vitro exposure to Mycoplasma gallisepticum. AB - Mycoplasma gallisepticum (MG) was used to expose chicken peripheral blood lymphocytes (PBLs), red blood cells (RBCs), heterophils, and chicken tumor cells (MSB-1 and HD-11 cells). Incubation of PBLs with MG for 3 hr resulted in extensive clumping of lymphocytes. Incubation of the MSB-1 cells with MG also caused clumping of the cells, with many of the cells showing perforations and others showing capping of the surface projections. Incubation of RBCs with MG resulted in an altered cell surface morphology, a decrease in cell size, and perforation. There were no discernible changes on the surface of the heterophils and the HD-11 cells. However, the HD-11 cells appeared to have a decreased ability to attach to the surface of the plastic and to have a decreased ability to respond to chemoattractant fMLP after 24 hr of incubation. These results suggest that, under the conditions used, MG caused certain damage to peripheral blood cells and a significant decrease in chemotactic response in the HD-11 cells. PMID- 15529969 TI - Construction and characterization of Marek's disease viruses having green fluorescent protein expression tied directly or indirectly to phosphoprotein 38 expression. AB - Marek's disease (MD) is caused by Marek's disease virus (MDV), a highly cell associated alphaherpesvirus. MD is primarily characterized by lymphocyte infiltration of the nerves and the development of lymphomas in visceral organs, muscle, and skin. MDV encodes two phosphoproteins, pp24 and pp38, that are highly expressed during lytic infection. These proteins were initially identified in MDV induced tumors but are now known to be linked primarily to MDV lytic infection. Despite the recent characterization of a pp38 deletion mutant MDV, the functions of these phosphoproteins remain unknown. The goal of this work was to construct recombinant MDVs having direct fusions of a marker gene, the green fluorescent protein (GFP), to pp38 in order to study the expression patterns and localization of this protein during stages of MDV infection. We report the construction of two recombinant viruses, one having the enhanced green fluorescent protein (eGFP) fused in-frame to the pp38 open reading frame (ORF) (RB1Bpp38/eGFP) and the other having soluble-modified GFP (smGFP) downstream but out-of-frame with pp38 (RB1Bpp38/smGFP). During construction of RB1Bpp38/eGFP, an ORF located downstream of pp38 (LORF12) was partially deleted. In RB1Bpp38/smGFP, however, LORF12 and its immediate 5' upstream sequence was left intact. This report describes the construction, cell culture, and in vivo characterization of RB1Bpp38/eGFP and RB1Bpp38/smGFP. Structural analysis showed that the virus stocks of RB1Bpp38/eGFP and RB1Bpp38/smGFP had incorporated the GFP cassette and were free of contaminating parent virus (RB1B). Moreover, RB1Bpp38/eGFP and RB1Bpp38/smGFP contained two and three and four and five copies of the 132-bp repeats, respectively. Expression analysis showed that the transcription of genes in RB1Bpp38/eGFP-and RB1Bpp38/smGFP-infected chicken embryo fibroblasts (CEFs) were similar to RB1B-infected CEFs, with the notable exception of deletion of a LORF12 specific transcript in RB1Bpp38/ eGFP-infected cells. In CEFs, RB1Bpp38/eGFP and RB1Bpp38/smGFP showed comparable one-step growth kinetics to parental virus (RB1B). RB1Bpp38/eGFP and RB1Bpp38/smGFP, however, showed quite distinct growth characteristics in vivo. Two independent clones of RB1Bpp38/eGFP were highly attenuated, whereas RB1Bpp38/smGFP exhibited pathogenesis similar to parent virus and retained oncogenicity. Our results suggest that the RB1Bpp38/eGFP phenotype could be due to an interference with an in vivo-specific pp38 function via GFP direct fusion, to the deletion of LORF12, or to a targeting of the immune response to eGFP. Because deletion of pp38 was recently found not to fully attenuate very virulent MDV strain MD-5, it is possible that deletion of LORF12 may be at least partially responsible for the attenuation of RB1Bpp38/eGFP. The construction of these viruses and the establishment of cell lines from RB1Bpp38/smGFP provide useful tools for the study of MDV lyric infection in cell culture and in vivo, in studies of the reactivation of MDV from latency, and in the functional analysis of LORF12. PMID- 15529971 TI - Pathogenesis of chicken anemia virus: comparison of the oral and the intramuscular routes of infection. AB - The events during the pathogenesis of chicken anemia virus (CAV) infection following intramuscular (IM) and oral inoculation were further elucidated and compared by sequential clinical, pathologic, and morphometric histopathologic evaluations, and by sequential determination of CAV genome concentrations in different organs. Specific-pathogen-free chickens were inoculated by IM or oral routes with the same dose (2 x 10(6) mean tissue culture infective dose [TCID50]) of CAV isolate 03-4876 at 1 day of age. Weights and hematocrits were obtained at 7, 10, 14, 18, 21, 25, and 28 days postinoculation (DPI). Seven birds from each group were necropsied at 7, 10, 14, and 28 DPI, and samples of thymus, Harderian gland, and cecal tonsils (CT) were obtained for histopathologic examination and CAV genome quantification by real-time polymerase chain reaction. Peak CAV genome concentrations were detected in the thymus at 10 and 14 DPI in the IM and orally infected chickens, respectively. High CAV DNA concentrations were maintained throughout the experimental period until 28 DPI, despite specific seroconversion occurring by 14 DPI in the IM-inoculated chickens. CAV was isolated from both orally and IM-infected chickens 28 DPI. Peak CAV genomes in the thymuses of IM and orally infected chickens coincided with peak lymphocyte depletion in these organs. Lymphocyte repopulation of the thymus occurred by 28 DPI in spite of the presence of the virus in the organs of both infected chicken groups. CAV genomes were detected in the CT, but histopathologic changes were not observed. Compared with the IM route of infection, orally infected chickens did not show apparent signs of illness. Clinical parameters, including reduction of weight gains and hematocrits, and gross and histopathologic changes were delayed and less severe in the orally inoculated chickens. This was concurrent with a delay in accumulation of CAV genomes in the thymus of these chickens. PMID- 15529972 TI - Mycoplasma meleagridis-induced lesions in the tarsometatarsal joints of turkey embryos. AB - Mycoplasma meleagridis (MM) has the ability to cause bone deformity in turkey poults. However, few pathological lesions have been described and no evidence of MM-induced damage to the bones has been shown. In this study, 17-day-old turkey embryos were inoculated with MM into the allantoic cavity. On the 27th day, eight of the 22 embryos presented with curved toes. Scanning electron microscopy of the tarsometatarsal joints showed fissures in the cartilage. Histological sections of the joints revealed only the infiltration of cells with eosinophilic granules. Immunohistochemical staining (IHS) showed the presence of MM in the aggregates of the bone marrow cells and the cells with eosinophilic granules. Some of these cells were harvested by laser capture microdissection (LCM), lysed, and used as template DNA. With a pair of MM-specific primers in a conventional polymerase chain reaction (PCR), a gene product was amplified, and it comigrated with the MM DNA, which indicates that these captured cells contained MM DNA. Thus, this research shows that inoculation of MM into the turkey embryos produced joint lesions and caused cellular infiltration within the bones. PMID- 15529973 TI - Genetic variation of viral protein 1 genes of field strains of waterfowl parvoviruses and their attenuated derivatives. AB - To understand the genetic variations between the field strains of waterfowl parvoviruses and their attenuated derivatives, we analyzed the complete nucleotide sequences of the viral protein 1 (VP1) genes of nine field strains and two vaccine strains of waterfowl parvoviruses. Sequence comparison of the VP1 proteins showed that these viruses could be divided into goose parvovirus (GPV) related and Muscovy duck parvovirus (MDPV) related groups. The amino acid difference between GPV- and MDPV-related groups ranged from 13.1% to 15.8%, and the most variable region resided in the N terminus of VP2. The vaccine strains of GPV and MDPV exhibited only 1.2% and 0.3% difference in amino acid when compared with their parental field strains, and most of these differences resided in residues 497-575 of VP1, suggesting that these residues might be important for the attenuation of GPV and MDPV. When the GPV strains isolated in 1982 (the strain 82-0308) and in 2001 (the strain 01-1001) were compared, only 0.3% difference in amino acid was found, while MDPV strains isolated in 1990 (the strain 90-0219) and 1997 (the strain 97-0104) showed only 0.4% difference in amino acid. The result indicates that the genome of waterfowl parvovirus had remained highly stable in the field. PMID- 15529974 TI - A prospective study of management and litter variables associated with cellulitis in California broiler flocks. AB - Cellulitis has emerged as an economically important disease of broiler chickens. The impact of environmental risk factors on the incidence of cellulitis has not been evaluated in the United States. Escherichia coli (E. coli), the causative agent, is introduced through skin scratches during the grow out. Our previous work suggested that the litter was an important reservoir for cellulitis associated E. coli. We hypothesized that factors contributing to a positive environment for E. coli growth would increase the opportunity for exposure of a broiler to an infectious dose of E. coli, capable of initiating a cellulitis lesion. This prospective study of 304 flocks on five farms from two integrated broiler companies was conducted to determine the effect of environmental factors on the prevalence of cellulitis in California broiler flocks. Environmental variables included temperature, wind velocity, and relative humidity (RH) at the litter surface. Litter variables measured included E. coli and total gram negative bacteria load (colony forming units/g dry matter), water activity, and pH. Management variables such as clean out, the number of flocks reared on the same litter (litter run, LR), and downtime (DT) between flocks were also evaluated. Cellulitis ranged from 0.197% to 6.04%. Significant associations were identified using linear regression between farm, LR, DT, ambient temperature during the brooding period, gram-negative bacteria load in the litter during the brooding period, RH mid-grow out, and E. coli load late in the grow out. The significant variation in the rate of cellulitis between farms combined with the strong association of LR and DT with cellulitis demonstrated that management choices were highly influential in this disease syndrome. Based on these data and our previous findings, managers would be advised to increase DT between flocks and perform a total clean out of the house when a flock processes with a high incidence of cellulitis. PMID- 15529975 TI - Development of a multiplex reverse transcription-polymerase chain reaction diagnostic test specific for turkey astrovirus and coronavirus. AB - A multiplex reverse transcription-polymerase chain reaction (RT-PCR) assay was developed for the simultaneous detection of two enteric viruses of poultry: turkey enteric coronavirus (TCV) and turkey astrovirus (TAstV). PCR primers were designed to conserved regions within the nucleocapsid gene of TCV and to the polymerase gene of TAstV-2. The primer pairs were successfully used in a multiplex RT-PCR to detect nucleic acid of TAstV-2 and TCV. The test was optimized for use with intestines/feces from naturally infected turkeys. The primers were specific and did not amplify other common RNA or DNA avian viruses. The detection limit was determined to be 10 ng of RNA used as starting template. The use of this specific test allows the rapid and early diagnosis of two financially costly viruses affecting the commercial turkey industry. PMID- 15529976 TI - Genetic characterization of very virulent infectious bursal disease viruses from Latin America. AB - Very virulent infectious bursal disease viruses (vvIBDVs) were detected in phenol inactivated bursal samples obtained from Brazil, the Dominican Republic, and Venezuela. After nucleotide sequence analysis of the hypervariable region of VP2 gene, the vvIBDVs from Brazil and Venezuela exhibited all of the 14 nucleotide changes that are conserved in the European UK-661 and most other vvIBDV strains. However, the vvIBDV from the Dominican Republic presented 11 nucleotide changes that are conserved in vvIBDV strains. After phylogenetic analysis, the Latin American strains were found to be related to other vvIBDV strains from Europe, Asia, and Africa. However, Brazilian and Dominican vvIBDVs clustered in two separate subgroups, while the vvIBDVs from Venezuela were closely related to other strains from other parts of the world. By deduced amino acid sequence, the three conserved amino acid residues in vvIBDV strains (222 Ala, 256 Ile, and 294 Ile) were confirmed in the Latin American viruses, and one amino acid change (300 Ala) was unique to all vvIBDVs from the Dominican Republic. The occurrence of this change in the Dominican vvIBDVs may have an impact in their antigenic makeup. Results of this study indicate that the vvIBDVs detected in Latin America are genetically similar to IBDV strains from other parts of the world. However, vvIBDVs from Venezuela were more similar to the vvIBDV strains from Europe and Asia. Of all the samples analyzed, vvIBDVs from Brazil and the Dominican Republic exhibited more genetic changes. These changes may have emerged as a result of the different management practices and environmental conditions present in each particular geographic area. PMID- 15529977 TI - Descriptive study of California egg layer premises and analysis of risk factors for Salmonella enterica serotype enteritidis as characterized by manure drag swabs. AB - This cross-sectional, double-blind study reports the prevalence of Salmonella enterica serotype enteritidis (SE) on California egg layer premises using single vs. pooled manure drag swabs and presents a description of egg production and management systems in the state and an initial analysis of risk factors for SE. The study included 91% of all known eligible egg premises in California, representing the majority of eggs produced in the state. The overall prevalence of SE on California egg layer premises was 10.5%, while 1.1% of all rows sampled were positive for SE. The percentage of positive rows for SE on any premises never exceeded 25% of the 16 swabs collected per premises. A description of egg production and management on California egg layer premises is presented. Statistically significant associations for SE were not evident and were limited because of sample size and the low prevalence of SE on California egg layer premises. Several biological and management factors, such as flock health, stage of production, manure management, ventilation, and watering systems, show trend associations with premises positive for SE and require further investigation. Manure drag swabs serve as a useful tool to validate the core components of an egg quality assurance program for SE based on process control principles. PMID- 15529978 TI - Molecular characterization of Mycoplasma gallisepticum isolates from turkeys. AB - Mycoplasma gallisepticum was isolated from several turkey flocks at different locations in the United States that were clinically affected with respiratory disease. Five of these isolates from four series of outbreaks had patterns similar to the 6/85 vaccine strain of M. gallisepticum by random amplified polymorphic DNA (RAPD) analysis using three different primer sets, whereas with a fourth primer set (OPA13 and OPA14), only two of the isolates were similar to 6/85. Results obtained by sequencing portions of the pvpA, gapA, and mgc2 genes and an uncharacterized surface lipoprotein gene indicated that the field isolates had DNA sequences that ranged from 97.6% to 100%, similar to the 6/85 results. In some of the outbreaks there was an indirect association with the presence of commercial layers in the area that had been vaccinated with this vaccine strain, but there was no known close association with vaccinated birds in any of the outbreaks. Turkeys were challenged with two of the field isolates and with 6/85 vaccine strain. Turkeys challenged with the field isolates developed respiratory disease with airsacculitis and a typical M. gallisepticum antibody response, whereas birds challenged with 6/85 developed no respiratory signs or lesions and developed only a weak antibody response. Although these isolates were very similar to the 6/85 vaccine strain, it was not possible to prove that they originated from the vaccine strain-it is possible that they could be naturally occurring field isolates. PMID- 15529979 TI - Pathologic findings in red-tailed hawks (Buteo jamaicensis) and Cooper's hawks (Accipiter cooper) naturally infected with West Nile virus. AB - Carcasses of 13 red-tailed hawks (RTHAs) and 11 Cooper's hawks (COHAs) were tested for West Nile virus (WNV) using WNV-specific reverse transcriptase polymerase chain reaction (RT-PCR) on fresh brain tissue and WNV-specific immunohistochemistry (IHC) on various organs. Ten COHAs (91%) and 11 RTHAs (85%) were positive for WNV RNA by RT-PCR. All 11 COHAs (100%) and 10 RTHAs (77%) were positive for WNV antigen by IHC. A triad of inflammatory lesions, including chronic lymphoplasmacytic and histiocytic encephalitis, endophthalmitis, and myocarditis, was common in both species. In COHAs, the heart (54%), cerebrum (50%), and eye (45%) were the organs that most commonly contained WNV antigen. The amount of WNV antigen was usually small. In RTHAs, the kidney (38%), cerebrum (38%), cerebellum (38%), and eye (36%) were the organs most commonly containing WNV antigen. Unlike COHAs, larger amounts of WNV antigen were present in the cerebrum of RTHAs. WNV antigen was detected in similar cell populations in both species, including neurons of brain, spinal cord, and retina, pigmented epithelial cells of the retina, epithelial cells of renal medullary tubules, cardiomyocytes, endothelial cells and smooth muscle cells of arteries, dendritic cells of splenic lymph follicles, exocrine pancreatic cells, adrenal cells, and keratinocytes of the skin. The study presents strong evidence that WNV can cause a chronic fatal disease in RTHAs and COHAs. The lesion distribution of WNV infection in both species is variable, but inflammatory lesions are common, and a triad of lesions including encephalitis, myocarditis, and endophthalmitis is indicative of WNV infection in both species. PMID- 15529980 TI - S1 and N gene analysis of avian infectious bronchitis viruses in Taiwan. AB - The disease caused by infectious bronchitis virus (IBV) produces great economic for the poultry industry. The purpose of this study is to investigate the molecular epidemiology of IBV in Taiwan. An old IBV strain isolated in 1964 and another 31 strains isolated from 1991 to 2003 were selected for N-terminal S1 gene analysis. Based on their phylogenetic tree, 13 strains were selected for sequencing the entire S1 and partial nucleocapsid (N) genes. The results indicated that Taiwanese IBV strains could be divided into two distinct lineages, Taiwan Group I and Taiwan Group II, with one Massachusetts strain and one Chinese strain. No recombination was found between H120 and the Taiwanese strains in the S1 gene. However, the S1 gene showed a noticeably higher divergence than the N gene. The phylogenetic trees constructed from the S1 and N genes indicate that intergenic recombination has occurred. Since most local strains are in Taiwanese clusters, developing vaccines from local strains is necessary for IBV control in Taiwan. PMID- 15529981 TI - The occurrence and distribution of Salmonella enteritidis and other serovars on California egg laying premises: a comparison of two sampling methods and two culturing techniques. AB - Between the summer of 1998 and the winter of 2000, Salmonella analysis was performed on 2128 single and 532 pooled manure drag swabs obtained from 133 California commercial egg laying farms. The isolation of Salmonella from all rows and from all flocks using single or pooled swabs was 80% and 92%, respectively. Hence, there was no statistical difference between single vs. pooled swabs in terms of identifying Salmonella on a row or flock basis. A total of 14 serogroups comprising 44 serotypes were isolated from 123 of 133 farms. When the top 10 serotypes were considered, there was no significant difference in the range of serotypes isolated by the two culturing methods. The overall S. enteritidis prevalence for California flocks was 10.5% (14/133). The overall row prevalence for S. enteritidis for all the farms was 1.1% (24/2128), and the overall pool prevalence was 2.4% (13/532). Sixty percent (12/20) of the S. enteritidis isolates from the positive farms were phage type 4, and 40% (8/20) represented five other phage types (1, 6B, 7, 8, and 28). PMID- 15529982 TI - Expression of Escherichia coli antigens in Salmonella typhimurium as a vaccine to prevent airsacculitis in chickens. AB - Avian pathogenic Escherichia coli strains are associated with a variety of extraintestinal poultry diseases, including airsacculitis, colisepticemia, and cellulitis. A number of E. coli serotypes are associated with these diseases, although the most prevalent serotype is O78. Fimbrial proteins expressed by these strains appear to be important virulence factors, including type 1 fimbriae, P fimbriae, and curli. We have been working to develop an effective vaccine to protect chickens against these diseases. We have previously shown that an attenuated Salmonella typhimurium strain expressing O78 lipopolysaccharide provides protection against challenge with an O78 avian pathogenic E. coli strain. In this work, we have constructed an attenuated S. typhimurium that expresses both the O78 lipopolysaccharide and E. coli-derived type 1 fimbriae. In these studies, chickens were vaccinated at day of hatch and again at 2 wk of age. Birds were challenged at 4 wk of age. We found that the vaccine candidate provided significant protection against airsacculitis as compared to untreated controls or birds vaccinated with an attenuated S. typhimurium that did not express any E. coli antigens. In a separate experiment, challenged vaccinates showed significant weight gain compared to challenged nonvaccinates. We were not able to demonstrate protection against E. coli O1 or O2 serotype challenge, nor against challenge with wild-type S. typhimurium. PMID- 15529984 TI - Chicken primary enterocytes: inhibition of Eimeria tenella replication after activation with crude interferon-gamma supernatants. AB - A reproducible and original method for the preparation of chicken intestine epithelial cells from 18-day-old embryos for long-term culture was obtained by using a mechanical isolation procedure, as opposed to previous isolation methods using relatively high concentrations of trypsin, collagenase, or EDTA. Chicken intestine epithelial cells typically expressed keratin and chicken E-cadherin, in contrast to chicken embryo fibroblasts, and they increased cell surface MHC II after activation with crude IFN-gamma containing supernatants, obtained from chicken spleen cells stimulated with concanavalin A or transformed by reticuloendotheliosis virus. Eimeria tenella was shown to be able to develop until the schizont stage after 46 hr of culture in these chicken intestinal epithelial cells, but it was not able to develop further. However, activation with IFN-gamma containing supernatants resulted in strong inhibition of parasite replication, as shown by incorporation of [3H]uracil. Thus, chicken enterocytes, which are the specific target of Eimeria development in vivo, could be considered as potential local effector cells involved in the protective response against this parasite. PMID- 15529983 TI - Specific detection and typing of Mycoplasma synoviae strains in poultry with PCR and DNA sequence analysis targeting the hemagglutinin encoding gene vlhA. AB - Mycoplasma synoviae is a major pathogen of chickens and turkeys, causing economic losses to the poultry industry worldwide. In this study, we validated and applied polymerase chain reaction (PCR) and DNA sequence analysis on the N-terminal end of the hemagglutinin encoding gene vlhA as an alternative for the detection and initial typing of field strains of M. synoviae in commercial poultry. PCR primers were tested against isolates of M. synoviae from various sources along with other avian mycoplasma and other bacterial species. The vlhA gene-targeted PCR assay was highly specific in the identification of M. synoviae, with a detection limit of 4.7 x 10(2) color changing units/ml. DNA sequence analysis of amplified products was also conducted to validate the potential for typing M. synoviae strains using the N-terminal region of the vlhA gene. To evaluate the test, we applied the PCR assay to tracheal swabs collected from chickens challenged with M. synoviae strain K1968 and compared the results to the serologic detection. The PCR assay was also evaluated directly on tracheal samples collected from commercial layers. Overall, this vlhA gene-targeted PCR is a useful tool for detection and initial typing of M. synoviae and can be applied in the preliminary identification of M. synoviae isolates directly from clinical samples. PMID- 15529985 TI - Isolation and prevalence of Campylobacter in the reproductive tracts and semen of commercial turkeys. AB - Campylobacter is one of the most commonly reported bacterial causes of human foodborne infections in the United States, and epidemiologic evidence indicates that a significant proportion of human infections result from the improper preparation of poultry products. Campylobacter frequently colonizes the avian intestinal tract, but recent research indicates that this organism can also colonize the avian reproductive tract and possibly contaminate eggs and subsequent offspring. The present studies were undertaken to determine the prevalence of Campylobacter in the reproductive systems of commercial turkeys. In the first study, pooled semen samples from seven commercial turkey farms were randomly collected by abdominal massage over a period of 13 wk. The pooled semen samples were serially diluted, and 0.1 ml of each dilution was plated on Campy Line agar and incubated at 42 C for 48 hr in a microaerophilic environment for enumeration of Campylobacter. Campylobacter was isolated from 57 of the 59 pooled semen samples, and levels ranged from below the limit of detection (<10(1)) to 1.6 x 10(6) cfu/ml of semen. In the second study, the reproductive tracts of 11 hens and 17 toms were aseptically excised, and the segments (female: vagina, shell gland, isthmus, magnum, and infundibulum; male: ductus deferens and testes) were swabbed with a dry cotton sterile swab. The swabs were incubated for 24 hr in Campylobacter enrichment broth, and 0.1 ml of the enriched sample solution was streaked onto Campy-Line agar plates and incubated at 42 C for 48 hr in a microaerophilic environment. Of the 11 hens sampled, Campylobacter was isolated from the vagina (10/11), the shell gland (7/11), the isthmus (8/11), the magnum (6/11), and the infundibulum (4/11). Of the 17 toms sampled, Campylobacter was isolated from the ductus deferens (8/17) and the testes (2/17). Campylobacter is present in the reproductive tracts and semen of commercial turkeys and may lead to vertical transmission of Campylobacter from the hen to the chick. PMID- 15529986 TI - Control of coccidiosis in turkeys with diclazuril and monensin: effects upon performance and development of immunity to Eimeria species. AB - The effects of diclazuril and monensin, when included in the feed of turkeys from 0 to 10 wk, upon performance and development of immunity to Eimeria species was investigated. Birds were initially inoculated with a low dose of oocysts of three species of Eimeria at 3, 5, 7, and 9 days of age in order to simulate a natural infection. Weight gain and feed intake from 0 to 6 wk of age was significantly greater in medicated birds compared with those that received no anticoccidial medication. Weight gain and feed intake from 6 to 10 wk was greater in birds that received diclazuril than in unmedicated birds. No differences in performance were evident after drug withdrawal from 10 to 16 wk. Immunity to Eimeria species developed by 10 wk in birds that received no anticoccidial medication but did not develop in those given diclazuril or monensin. PMID- 15529988 TI - Safety of Mycoplasma gallisepticum vaccine strain 6/85 after backpassage in turkeys. AB - The objective of this research was to evaluate the safety of the 6/85 strain vaccine strain of Mycoplasma gallisepticum in turkeys by backpassing the vaccine strain up to 10 times by contact infection in turkeys and challenging turkeys with the resulting backpassaged strain. The vaccine strain, however, did not spread to in-contact turkeys, and it was necessary to reisolate the organism before challenging turkeys for the next passage. The challenge strain, therefore, was one that had been backpassaged four times in turkeys, with a total in vivo time in turkeys of 66 days. The backpassaged 6/85 vaccine strain was no different in pathogenicity than the original vaccine strain, except that at 10 days postchallenge, it was isolated in higher numbers from air sacs. Both the original 6/85 vaccine strain and the backpassaged strain were apathogenic in turkeys, except for a slightly increased diameter of the tracheal mucosa at 10 days postchallenge; at 20 days postchallenge the tracheal mucosal thickness was no different from that of controls. PMID- 15529987 TI - Identification of a novel nephropathogenic infectious bronchitis virus in Israel. AB - A novel infectious bronchitis variant, designated as IS/885/00, associated with nephritis, was isolated from outbreaks in 23 broiler farms in Israel. The virus was first identified by reverse transcriptase-polymerase chain reaction and showed a distinct restriction fragment length polymorphism pattern from previously described Israeli isolates. Sequence analysis of the S1 gene and the deduced amino acid sequence revealed 97.2% protein similarity to genotype IS/ 720/99 and 71.6% similarity to the vaccine strain H120, the only strain permitted for use in this country. A database search in GenBank revealed a closely related isolate from Egypt, Egypt/Beni-Seug/01, with 96.6% similarity. Other published nephropathogenic infectious bronchitis virus strains/isolates shared less than 77% similarity with IS/885/00. A vaccine protection test in specific-pathogen free chicks indicated 91% protection to the trachea and only 25% protection to the kidneys in vaccinated birds challenged with IS/885/00. PMID- 15529989 TI - Inability of cecal microflora to promote reversion of viable nonculturable Campylobacter jejuni. AB - Campylobacter jejuni cells are able to enter a viable but nonculturable (VBNC) state when they are suspended in water. In the present experiments we inoculated day-of-hatch leghorn and broiler chicks with normal gut microflora and subsequently challenged these with high doses of VBNC C. jejuni. The objective was to determine if the pre-establishment of a normal gut flora would enable VBNC Campylobacter to recover, revert to the vibrionic form, and colonize the cecum. Day-of-hatch leghorn and broiler chicks were gavaged through the esophagus with 0.75 ml of a continuous-flow culture of normal cecal organisms. Two days after gavage, the same chicks were gavaged with 0.75 ml (greater than 10(9) colony forming units) of a VBNC suspension of C. jejuni. Seven days later, cecal contents were collected, serially diluted, and examined for the presence of viable culturable C. jejuni. Our results demonstrated that the VBNC C. jejuni cells were unable to revert to a vibrionic culturable form capable of colonizing the cecum. PMID- 15529990 TI - Phylogenetic analysis of the sigma 2 protein gene of turkey reoviruses. AB - The open reading frame of the S3 segment encoding the sigma2 protein of four turkey reovirus field isolates was analyzed for sequence heterogeneity. The turkey reoviruses we present here have a 97% amino acid identity to turkey NC 98. The S3 nucleotide and amino acid sequence similarity was < or =61% and 78%-80%, respectively, when compared to the chicken reovirus isolates. Comparison of amino acid sequences from chickens and turkeys with that of a duck isolate revealed a 53% and 55% similarity, respectively. Phylogenetic analyses, based on both nucleotide and amino acid sequence, resulted in three major groups among the avian reoviruses; these groups were clearly separated by species. The results of this study provide further evidence, based on the deduced sigma2 sequence, that turkey reoviruses form a distinct, separate group relative to chicken and duck isolates. In addition, as a result of the limited sequence identity with their avian counterparts, turkey reoviruses could potentially be considered a separate virus species within subgroup 2 of the Orthoreovirus genus. PMID- 15529991 TI - Comparative efficacy of enrofloxacin, oxytetracycline, and sulfadimethoxine for the control of morbidity and mortality caused by Escherichia coli in broiler chickens. AB - The purpose of the present study was to compare the ability of enrofloxacin, oxytetracycline, and sulfadimethoxine to reduce morbidity and mortality caused by Escherichia coli (colibacillosis) in broiler chickens. The chickens were raised in 80 pens (20 birds per pen) with 20 pens representing each treatment group under simulated commercial conditions that produced a colibacillosis challenge scenario. Each group of 20 randomized pens (replicates) was given one of four water treatments. Chickens that received enrofloxacin had significantly less mortality (P < 0.01), lower average gross pathology (colibacillosis) scores (P < 0.01), and better feed-conversion ratios (P < 0.05) than did chickens that received either oxytetracycline or no medication. Chickens that received enrofloxacin had significantly less mortality and lower pathology scores than those that received sulfadimethoxine and numerically lower feed conversion than the sulfadimethoxine group. Results from the present study show that enrofloxacin is superior to oxytetracycline and sulfadimethoxine for the control of morbidity and mortality caused by E. coli in broiler chickens. Our findings will help veterinarians choose and prescribe the most efficacious antimicrobial when treating colibacillosis. PMID- 15529992 TI - Immunization of chickens with VP2 protein of infectious bursal disease virus expressed in Arabidopsis thaliana. AB - Transgenic plants represent a safe, effective, and inexpensive way to produce vaccines. The immunogenicity of VP2 protein of an infectious bursal disease (IBD) virus variant E isolate expressed in transgenic Arabidopsis thaliana was compared with a commercial vaccine in specific-pathogen-free broiler chickens. The VP2 coding sequence was isolated and integrated into A. thaliana genome by Agrobacterium tumefaciens-mediated transformation. Soluble VP2 expressed in transgenic plants was used to immunize chickens. Chickens receiving oral immunization with plant-derived VP2 at 1 and 3 wk of age had an antibody response using enzyme-linked immunosorbent assay and 80% protection against challenge infection at 4 wk. Chickens primed with a commercial vaccine at 1 wk followed by an oral booster with VP2 expressed in plants at 3 wk of age showed 90% protection. Chickens immunized with a commercial vaccine at 1 and 3 wk showed 78% protection. Results supported the efficacy of plant-produced VP2 as a vaccine against IBD. PMID- 15529993 TI - Survival of exotic Newcastle disease virus in commercial poultry environment following removal of infected chickens. AB - During the first weeks of 2003, after exotic Newcastle disease (END) was confirmed in commercial layer flocks in Southern California, it became apparent that the virus survival information in the literature varied widely and was difficult to extrapolate to current local conditions. The END Task Force used the information available in the literature and the recommendations of research scientists to establish protocols for safely handling manure from infected and depopulated premises. In an attempt to gain more applicable knowledge in the management of contaminated poultry manure in the course of the END outbreak, this virus survival study was designed and implemented. Environmental drag swabs were tested for END virus from two of the early-infected commercial ranches that consisted of several houses following immediate removal of the infected flocks. A total of 293 samples, composed of 168 manure drag swab pools, 72 dropping board swab pools, and 38 compost swab pools from 3 houses (ranch 1), and 180 manure belt scraper swab pools from ranch 2 were analyzed for ND virus isolation and characterization for 21 consecutive days postdepopulation. Thirteen manure drag swab pools (from houses 1 and 3) and two manure dropping board swab pools (from house 3) collected from ranch 1 were positive for END virus at 0, 1, 2, 3, 4, 7, 10, 12, and 16 days postdepopulation. No END virus was isolated after the 16th day following depopulation from any of the samples. All samples from ranch 2 were negative during the entire observation period. PMID- 15529994 TI - High mortality in egg layers as a result of necrotic enteritis. AB - A new facility was designed to hold 1.8 million birds in 10 houses; chickens were placed in five of the houses, and the remaining five houses were under construction when this outbreak occurred. An increase in mortality was reported in five houses; however, mortality in house 7 was quite high. Well-fleshed birds were suddenly found dead without a significant drop in egg production. The middle and distal intestines were distended with gas, congested, thin walled, atonic, and bluish or pale in color with sloughed mucosa in some places. Necrotic enteritis was diagnosed as the cause of increased mortality. The ingesta in the crop occasionally contained flies. The 4-wk mortality in house 7 was 6.55% with a loss of 10,898 chickens. The 4-wk mortality rate in the other houses ranged from 0.54% to 1.98%. The houses affected with necrotic enteritis were treated for coccidiosis with amprolium because low numbers of the oocysts were present in the intestinal specimens of some of the chickens. Household bleach was added to the water at a dilution of one part bleach to 1040 parts water to control bacterial contamination. The fly (Musca domestica) population was out of control. Clostridium perfringens was isolated from the alcohol-washed macerated flies caught from houses 4 and 7. Dead flies were often seen in the feed troughs. The chickens may possibly have had C. perfringens infection as a result of consumption of dead flies or their secretions/excretions. The alcohol-washed, macerated, clarified fly extract from the affected houses caused death in 11 inoculated mice and paralysis in one mouse. Similarly, illness and mortality were present in four mice inoculated with clarified intestinal contents. The bacterium isolated on anaerobic culture was identified as C. perfringens by polymerase chain reaction. The disease was brought under control after straw was added and mixed in with the litter. As a result, the litter temperature increased, causing a decrease in the fly population. This study suggests that flies in the poultry houses acted as mechanical transmitters of C. perfringens and that the development of necrotic enteritis was by ingestion of bacteria present in the flies and their secretions/excretions. PMID- 15529995 TI - Venereal colibacillosis (acute vaginitis) in turkey breeder hens. AB - Two flocks of turkey breeders experienced an increased mortality and high culling rate in the first weeks of egg production. The majority of dead and culled hens had cheesy core in the cloaca and vagina. Postmortem examination revealed fibrinous pseudomembranes in the vagina and cloaca. The thickness of these membranes posed an obstruction to egg passage leading to internal laying and egg peritonitis. Swabs from cloaca and vagina produced numerous colonies of only E. coli. Investigations of this unusual vaginitis showed that these two flocks had a higher number of immature hens with present hymens, and insemination crews mistakenly inseminated all hens in which they were able to evert the cloaca. Breaking the hymen with an insemination pipette created a wound and developed extensive infection with E. coli bacteria. PMID- 15529996 TI - Colobomas of the iris in a flock of rosecomb bantam chickens. AB - A defect in the pupil shape ofrosecomb bantam chickens was noticed by a breeder. The pupil in affected birds appeared to be elongated at the lower margin and consistent with a coloboma of the iris. Upon examination of all birds in the flock, the defect was found in 26% of females and 9% of males. Examination of breeders older than 1 yr led to the discovery ofcolobomas, although there were not enough birds to accurately determine the prevalence of the problem for previous generations. Affected birds were examined for other lesions, and none were found. Specific-pathogen-free (SPF) eggs were incubated and hatched with eggs from the affected flock as sentinels. No SPF chicks developed colobomas of the iris, while 37% of the bantams in the same hatch were affected. This experiment eliminated many possible infectious agents and incubator or brooder malfunction as causes of this eye lesion. An analysis of the flock pedigree suggested that the defect might be inherited and might be related either to feather color or to gender. No statistically significant correlation could be made between feather color and the iris colobomas. However, the trait was twice as likely to occur in females as in males, which suggests that it is a sex influenced trait. Based on the determination that this was an inherited trait, a breeding strategy to reduce the prevalence of the lesion in which affected birds were removed from the breeding flock was developed and followed for 2 yr. The prevalence of colobomas was 22% in females and 2.9% in males after the first year and 7.8% and 0.8% in females and males, respectively, after the second year. PMID- 15529997 TI - The highly pathogenic avian influenza A (H7N7) virus epidemic in The Netherlands in 2003--lessons learned from the first five outbreaks. AB - Clinical signs and gross lesions observed in poultry submitted for postmortem examination (PME) from the first five infected poultry flocks preceding the detection of the primary outbreak of highly pathogenic avian influenza (HPAI) of subtype H7N7 during the 2003 epidemic in the Netherlands are described. The absence of HPAI from the Netherlands for more than 75 yr created a situation in which poultry farmers and veterinary practitioners did not think of AI in the differential diagnosis as a possible cause of the clinical problems seen. Increased and progressive mortality was not reported to the governmental authorities by farmers or veterinary practitioners. It took 4 days from the first entry of postmortem material to notify the governmental authorities of a strong suspicion of an AI outbreak on the basis of a positive immunofluoresence test result. The gross lesions observed at PME did not comply with the descriptions in literature, especially the lack of hemorrhagic changes in tissues, and the lack of edema and cyanosis in comb and wattles is noted. The following lessons are learned from this epidemic: a) in the future, increased and progressive mortality should be a signal to exclude AI as cause of disease problems on poultry farms; b) intensive contact between the veterinary practitioner in the field and the veterinarian executing PME is necessary to have all relevant data and developments at one's disposal to come to a conclusive diagnosis; c) in an anamnesis, reporting of high or increased mortality should be quantified in the future (number of dead birds in relation to the number of birds brought to the farm to start production, together with the timing within the production cycle), or else this mortality cannot be interpreted properly; d) if clinical findings such as high mortality indicate the possibility of HPAI, the pathologist should submit clinical samples to the reference laboratory, even if PME gives no specific indications for HPAI; e) the best way to facilitate early detection of an HPAI outbreak is to have the poultry farmer and/or veterinary practitioner immediately report to the syndrome-reporting system currently in operation the occurrence of high mortality, a large decrease in feed or water intake, or a considerable drop in egg production; f) in order to detect low pathogenic avian influenza infections that could possibly change to HPAI, a continuous serologic monitoring system has been set up, in which commercial poultry flocks are screened for antibodies against AI virus of subtypes H5 and H7. PMID- 15529998 TI - Hexamita meleagridis (Spironucleus meleagridis) infection in chukar partridges associated with high mortality and intracellular trophozoites. AB - An outbreak of infectious catarrhal enteritis, associated with the flagellated protozoan Spironucleus meleagridis (syn. Hexamita meleagridis), is reported from a commercial flock of chukar partridges in California. The disease affected birds between the ages of 4 and 6 wk and resulted in diarrhea, listlessness, depression, and high mortality. Concurrent infection with other intestinal pathogens, including Cryptosporidia, group E Salmonella, long-segmented filamentous microorganisms (LSFMOs), and Rotavirus-like virus particles, was found in some but not all affected birds. Dermatitis of the face, shanks, and feet, suggestive of B-complex vitamin deficiency, was present in most affected birds as well. Flagellated protozoan parasites could be found in the lumen of the duodenum and jejunum and in the intestinal crypts. In some cases the flagellates were wedged between epithelial cells or were located intracellularly within cells of the mucosal epithelium and the intestinal lamina propria. PMID- 15529999 TI - Histomoniasis in the bursa of fabricius of chickens. AB - Histomoniasis was diagnosed in a flock of 6-wk-old commercial chickens. Clinical signs included depression, stilted gait, inappetence, and a slight increase in mortality. At necropsy, there were pale-yellow to dark-gray circular and depressed necrotic lesions in the liver. The ceca were enlarged and impacted with caseous cores. Cecal worms were not observed either at necropsy or on histopathology. Histomonads were demonstrated microscopically within the bursa of Fabricius in addition to the liver, ceca, and spleen. This is the first report of the presence of histomonads in the bursa of Fabricius in commercial chickens. PMID- 15530000 TI - Bacteriologic findings in ostrich (Struthio camelus) eggs from farms with reproductive failures. AB - From January 2001 to December 2002, 543 ostrich eggs were submitted for bacteriologic investigation. The eggs were laid by 387 domesticated ostriches that suffered fertility disorders and that came from 44 farms located in different areas of Northern and Central Italy. Microbiologic investigations showed bacterial isolation in 105 (19.3%) of 543 eggs examined, with a high prevalence of enterobacteria from albumen and yolk. In only a few cases did bacterial isolation result from yolk or albumen alone. An antibiotic sensitivity test was conducted on isolates by the Kirby-Bauer disc diffusion method. This is the first report regarding the microbiologic status of eggs from ostrich farms located in different Italian regions. PMID- 15530001 TI - Natural microbial UV radiation filters--mycosporine-like amino acids. AB - Ozone depletion by anthropogenic gases has increased the atmospheric transmission of solar ultraviolet-B radiation (UV-B, 280-315 nm). There is a logical link between the natural defenses of terrestrial and marine organisms against UV radiation and the prevention of UV-induced damage to human skin. UV light degrades organic molecules such as proteins and nucleic acids, giving rise to structural changes that directly affect their biological function. These compounds offer the potential for development of novel UV blockers for human use. The biological role of mycosporine-like amino acids (MAAs) and scytonemin as a defense against solar radiation in organisms, together with their structure, synthesis, distribution, regulation and effectiveness, are reviewed in this article. This review points to the role of MAAs as a natural defense against UV radiation. PMID- 15530004 TI - Improvement of transformation and electroduction in avermectin high-producer, Streptomyces avermitilis. AB - Factors affecting the PEG-mediated transformation and electrotransformation of Streptomyces avermitilis protoplasts, an industrial avermectin high-producer, were evaluated. The maximum protoplast transformation efficiency under optimum conditions with PEG was 3 x 106 transformants per microg plasmid pIJ702 DNA. The efficiency of electrotransformation with the same plasmid the intact cells grown in medium with 0.5 mmol/L CaCl2, suspended in buffer with 0.5 mol/L sucrose +1 mmol/L MgCl2, and pulsed at an electric field strength of 10 kV/cm, 800 ohms, 25 microF, was of 2 x 10(3) transformants per microg DNA. When the cells were electroporated after mild lysozyme-treatment, the efficiency was up to 10(4) transformants per microg DNA. Electroporation of protoplasts and germlings had a lower efficiency (10(2) transformants per microg DNA). We report that electroporation under optimum conditions can be used for direct transfer of nonconjugative plasmid pIJ699 between two different Streptomyces species, S. avermitilis and S. lividans. PMID- 15530003 TI - Molecular epidemiology of group B streptococcal infections. AB - Streptococcus agalactiae (GBS) is a causative agent of sepsis and meningitis in newborns and diseases in pregnant women and nonpregnant adults. Various approaches, including both nongenetic and genetic techniques, are currently used for the study of epidemiology of GBS infections. In the present paper the different methods of molecular epidemiology of GBS infections are reviewed, and several novel approaches are introduced. The advantages and disadvantages of molecular methods are discussed and compared with traditional serotyping technique. The possible use of the molecular approaches for identification of different genetic lineages in GBS as well as for identification and control of the epidemiologically actual clones is discussed. PMID- 15530005 TI - Screening of lactic-acid bacteria from South African barley beer for the production of bacteriocin-like compounds. AB - Strains of Lactobacillus paracasei subsp. paracasei (strain ST11BR), L. pentosus (strain ST151BR), L. plantarum (strain ST13BR), and Lactococcus lactis subsp. lactis (strain ST34BR) producing bacteriocin-like peptides were isolated from barley beer produced in the Western, Northern and Eastern provinces of South Africa. The peptides (bacST11BR, bacST151BR, bacST13BR and bacST34BR) lost their activity after treatment with proteinase K, a proteinase, papain, chymotrypsin, trypsin, pepsin and pronase, but not when they were treated with alpha-amylase, suggesting that the peptides are not glycosylated. The peptides inhibited the growth of Lactobacillus casei, L. sakei, Pseudomonas aeruginosa, Escherichia coli and Enterococcus faecalis, but not Enterobacter cloacae, Lactobacillus bulgaricus subsp. delbrueckii, L. plantarum, L. salivarius, Listeria innocua, Staphylococcus aureus, Streptococcus uberis, S. agalactiae, S. caprinus and S. pneumoniae. Peptides bacST11BR and bacST13BR differed from the other 2 peptides by failing to kill Klebsiella pneumoniae and one of the E. coli strains. Peptides were stable after 2 h of incubation at pH 2.0-12.0, and after 90 min at 100 degrees C. When autoclaved (121 degrees C, 20 min), only bacST13BR lost its activity. The bacteriocin-like peptides were produced at a growth temperature of 30 degrees C, but not at 37 degrees C. PMID- 15530006 TI - Antimicrobial resistance of Enterococcus spp. isolates from raw beef and meat products. AB - E. faecalis (67%) and E. faecium (13.7%) were most frequently isolated among enterococci that contaminate cooled and frozen processed meat, follow-up heat treated meat products and unheated fermented dry salami. Most isolates of both species were resistant to cephalothin (95 and 83 %) and clindamycin (77 and 67%, respectively). Furthermore, E. faecalis and E. faecium isolates were resistant to erythromycin (44 and 72%), tetracycline (34.5 and 17.4%), and streptomycin (13.3 and 4.3%, respectively). Only a few of the isolates were resistant to ampicillin, ampicillin-sulbactam, chloramphenicol, and vancomycin while all isolates were susceptible to gentamicin, penicillin, and teicoplanin. During the production of heat-treated meat products, numbers of resistant isolates increased in spite of the decreasing enterococcal contamination of the samples. An opposite situation was found in the production of fermented dry salami. PMID- 15530002 TI - Molecular diagnostics of clinically important staphylococci. AB - Bacterial species of the genus Staphylococcus known as important human and animal pathogens are the cause of a number of severe infectious diseases. Apart from the major pathogen Staphylococcus aureus, other species until recently considered to be nonpathogenic may also be involved in serious infections. Rapid and accurate identification of the disease-causing agent is therefore prerequisite for disease control and epidemiological surveillance. Modern methods for identification and typing of bacterial species are based on genome analysis and have many advantages compared to phenotypic methods. The genotypic methods currently used in molecular diagnostics of staphylococcal species, particularly of S. aureus, are reviewed. Attention is also paid to new molecular methods with the highest discriminatory power. Efforts made to achieve interlaboratory reproducibility of diagnostic methods are presented. PMID- 15530007 TI - Conidia production of Beauveria sp. by solid-state fermentation for biocontrol of Ilex paraguariensis caterpillars. AB - Conidia production of Beauveria sp. strain LAG by solid-state fermentation (SSF) using blends of agro-industrial residues (residual potatoes and sugar-cane bagasse) was optimized with respect to cultivation conditions and the composition of substrate mixture in Erlenmeyer flasks and column-type bioreactor. With a blend of 60 % residual potatoes and 40 % sugar-cane bagasse the optimum conditions achieved were: incubation temperature 26 degrees C, initial substrate pH 6, inoculum concentration 10(7) conidia per g substrate; optimal initial moisture of the substrate was 70 % for Erlenmeyer flasks, in column-type bioreactor (with forced aeration) the optimal initial moisture of the substrate was 65 % with airflow of 60 mL/min. The highest production (1.07 x 10(10) conidia per g dry substrate) was achieved after a 10-d fermentation. The conidia were used in laboratory assays against Thelosia camina and Hylesia sp., caterpillars that are serious pests of mate plants. The mortality of T. camina was >90 % 10 d after spraying caterpillars with 1 mL conidia suspension at a concentration 10(5) 10(8)/mL. For Hylesia sp., the mortality was 70 %, 7 d after immersion in the conidia suspension containing 108 conidia per mL. Therefore, the Beauveria sp. LAG can be considered to be an important biocontrol instrument in the prospect of the Integrated Pest Management for mate plants. PMID- 15530008 TI - Soil microbial counts and identification of culturable bacteria in an extreme by arid zone. AB - Sixteen samples of two soil cores (about 550 and 180 cm in depth) were drilled at intervals in the lower reach of Heihe river basin (northwest of China) in order to illustrate soil microbial characteristics and diversity of culturable bacteria in an extreme by arid environment. Soil water content, organic matter, total nitrogen, pH, direct cell counts, and culturable microorganism counts were evaluated. The total cell concentration was 19-1120/microg (i.e. 0.19-11.2 x 10(8) per g) soil, the culturable bacteria count being 0.2-10.9 per microg (i.e. 2 x 10(5)-10.9 x 10(6) CFU/g) soil. The number of direct cell counts obtained by 4',6-diamidino-2-phenylindole-staining or the cound of culturable microbes after enrichment with different media were statistically significantly correlated with soil organic matters, total nitrogen content, soil water content and surface vegetation; this partly explained the larger number in the deeper first core than in the shallower one. As part of identification of 228 colonies isolated from the two cores, thirty-two were selected for 16S rDNA amplification, sequencing and molecular identification. These 32 isolates were affiliated to 5 major groups of bacteria: alpha-Proteobacteria, 5-Proteobacteria, gamma-Proteobacteria, the high G+C G+-bacteria, the low-G+C G- -bacteria, and the Cytophaga-Flexibacter Bacteroides group. Twenty-eight were rod- or short-rod shaped, which accounted for >87.5% of all species; only 4 of 32 species were cocci (<12.5%). PMID- 15530009 TI - Diversity of keratinophilic mycoflora in the soil of Agra (India). AB - Diversity of keratinophilic mycoflora in the soil of Agra was under observation for 1 year (July 2001-June 2002) and isolation of keratinophilic fungi was followed by the hair-baiting method. The frequency of occurrence of keratinophilic fungi in 284 soil samples collected from various hospitals, cattle yards, poultry farms, crop fields and playgrounds was determined, 204 samples (72%) having been found to be positive. A total of 33 species classified into 11 genera (Acremonium, Aspergillus, Chrysosporium, Emmonsia, Geomyces, Keratinophyton, Microsporum, Myceliophthora, Penicillium, Sporotrichum, Trichophyton) were encountered from the soil samples. Sporotrichum spp. were found to be the most dominant species followed by Trichophyton simii. The parameter of keratinophilic fungi found in the samples studied ranged from 62 to 80% where playgrounds yielded the maximum number of species (80%) while the least dominating soil was hospital soil (62%). Among all the baits used maximum fungi occurred on human hairs (82%) followed by chicken feather (74%), wool (61%) and the least on horns (45%). The spectrum of keratinophilic fungi isolated from different sites differed considerably according to the frequency of use by humans. PMID- 15530010 TI - Comparison of light and dark nitrogenase activity in selected soil cyanobacteria. AB - Frequency of heterocytes and nitrogenase activity (NA) under light and dark cultivation conditions was determined in 12 cyanobacterial strains isolated from various soil habitats. In spite of a high variability, significant differences in NA among the strains were found in response of light and dark cultivation. Relatively high NA (9.9-15.3 micromol/h C2H4 per g fresh mass) under light conditions and basal NA after 12 h of dark cultivation were detected in Anabaena, Nodularia, Tolypothrix, and 1 of Cylindrospermum strains. On the other hand, significantly lower NA (0.76-5.4 micromol/h C2H4 per g fresh mass) was found under light conditions in Trichormus, Nostoc and another Cylindrospermum strain; the activity completely disappeared after 12 h of dark cultivation. NA values were not directly related to the frequency of the heterocytes. The total NA of cyanobacterial colony was found to be probably independent of the number and/or position of heterocytes. Remarkable differences in NA between strains isolated from cultivated fields and strains originating from natural or non-cultivated soils were found. PMID- 15530011 TI - The effect of nalidixic acid on growth and reproductive events in nucleocytosolic and chloroplast compartments in the alga Scenedesmus quadricauda. AB - The courses of rRNA accumulation, DNA replication, and nuclear division were followed both in the chloroplast and the nucleocytosolic compartments during the cell cycle in synchronized populations of the chlorococcal alga Scenedesmus quadricauda. Control and nalidixic acid-treated cultures were compared. Nalidixic acid (150 mg/L) was added either at the beginning of the cell cycle or consecutively during the cell cycle to subcultures transferred into the dark. If the inhibitor was applied at the beginning of the cell cycle, chloroplast DNA did not replicate and nucleoids did not divide. Chloroplast division, however, was coordinated in a timely fashion with cytokinesis even under conditions of blocked chloroplast DNA replication. While the growth rate was slowed down, the courses of reproductive processes in the nucleocytosolic compartment were not affected and their timing and the number of rounds were coordinated with growth rate as in the control culture. The rate of cytosolic rRNA synthesis was lower but no apparent effect was seen on the amount of rRNA that accumulated during the cell cycle. In contrast, lower levels of chloroplast rRNA were found at the end of the cell cycle compared with the control culture. Experiments in which cells were transferred to the dark during the cell cycle showed that the inhibitor affected none of the reproductive events in the nucleocytosolic compartment. In the chloroplast compartment, DNA replication was inhibited in inhibitor-treated cultures, but was unaffected in controls. The chloroplast nucleoids themselves divided even in the presence of the inhibitor, reducing their DNA content to a level which corresponded to that in freshly formed control daughter cells. PMID- 15530012 TI - Detection of cefotaxime-resistant CTX-M-3 in clinical isolates of Serratia marcescens. AB - Strains of Serratia marcescens (isolated in a hospital during April and August 2000) resistant to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, streptomycin, tetracycline, and gentamicin were characterized. Out of a total of 34 clinical isolates 6 (17.6 %) exhibited the extended spectrum beta-lactamases (ESBL) resistance; they were also resistant to cefotaxime (minimum inhibitory concentration, MIC > or = 128 microg/mL) but susceptible to imipenem (MIC < or = 0.5 microg/mL). This multidrug resistance was shown to be transferred by a conjugative plasmid. Transconjugants revealed similar MIC profiles when compared to the parental strains. Isoelectric focusing revealed one major transferable beta-lactamase (pI 8.4) which was further identified as CTX-M-3 by PCR and gene sequencing. The presence of strains with this type of ESBL showed the evolution of bla genes and their dissemination among at least three species of the family Enterobacteriaceae isolated within a single hospital. The predominance of CTX-M type enzymes found in this area of Taiwan appeared to be similar to that described in Poland. PMID- 15530013 TI - Occurrence of endemic plasmids causing beta-lactam resistance in Enterobacteriaceae in children's university hospital in Munich. AB - Susceptibility of 62 clinical isolates of Enterobacteriaceae to 15 aminoglycosides, beta-lactams and fluoroquinolones was determined. The isolates originating from 3 intensive care units (neonatal, pediatric, and surgical) and the Department of Infant Internal Medicine of the Children's University Hospital City Center in Munich (Germany) were collected in August 1999, and March and October 2000. Transferability of antibiotic resistance from donors to their E. coli transconjugants was also demonstrated. The majority of isolates were resistant to ampicillin, cefoxitin, ceftriaxone, cefotaxime, ceftazidime and azthreonam but they were susceptible to cefepime, meropenem, aminoglycosides and fluoroquinolones. The occurrence of beta-lactamases and extended-spectrum beta lactamases (ESBL) was also shown. In August 1999 75% of isolates produced beta lactamases and 15% ESBL, in March 2000 95% of isolates produced beta-lactamases and 9% ESBL; in October 2000 all isolates produced beta-lactamases and only 5% produced ESBL. Plasmid DNA analysis in randomly chosen isolates and their transconjugants revealed the presence of plasmids ranging from 19 to 136 kb; in the majority of isolates a 120-kb plasmid was observed. Further analysis using restriction endonuclease suggested a dissemination and persistence of an endemic plasmid at all 4 wards of the large pediatric hospital in the City Center of Munich which may be responsible for resistance to beta-lactams among Enterobacteriaceae isolates. PMID- 15530014 TI - Urovirulence of Pseudomonas aeruginosa: planktonic cells vs. biofilm cells. AB - Planktonic and biofilm cells of a clinical urinary isolate of P. aeruginosa were compared in vitro for their ability to adhere to uroepithelial cells, interaction with macrophages, and for production of virulence factors like extracellular proteinase, elastase, hemolysin, phospholipase C and pyochelin. Biofilm cells showed increased adherence to UECs, which was coupled with reduced uptake and intracellular killing by macrophages. Overall there was a decrease in production of extracellular products by biofilm cells. Comparing the two cell forms for their ability to establish infection in an ascending model of acute pyelonephritis, significant enhancement of renal bacterial load, as well as more pronounced renal pathology developed with biofilm cells. PMID- 15530015 TI - The sensitivity of escherichia coli strains with K1 surface antigen and rods without this antigen to the bactericidal effect of serum. AB - The susceptibility of Escherichia coli strains with K1 surface antigen (K1+) and rods without this antigen (K1-) to the bactericidal action of normal bovine serum and human normal cord serum was determined. Seventy E. coli strains (35 K1+ and 35 K1-) were isolated from urine obtained from children with urinary tract infections. The strains investigated showed variable sensitivity to the bactericidal action of the sera. E. coli K1+ strains were characterized by lower sensitivity to bactericidal effect of the sera in comparison with K1- rods. The role of the particular mechanisms of complement activation in the process of killing of the E. coli strains was also determined. PMID- 15530016 TI - Effect of short-chain acids on the carboxymethylcellulase activity of the ruminal bacterium Ruminococcus albus. AB - The addition of 100-300 mmol/L of acetic, propionic, butyric or lactic acids (short-chain acids), or of acetic, propionic, and butyric acids (volatile fatty acids, VFA) mixtures increased the degradation of carboxymethyl cellulose (CMC) by R. albus (7.5 to 46 and 6 to 39 %, respectively). Differences among individual acids were observed at 300 mmol/L whereas VFA mixtures differed at 100 mmol/L. When assayed at the same concentration, CMCase activity was increased less by NaCl than by the short-chain acids, whereas ethylene glycol decreased the activity. Since osmolarity and/or ionic strength changes in the medium cannot completely account for the observed increases of carboxymethylcellulase (CMCase) activity, it is suggested that the anions of short-chain acids produce changes in the reaction media polarity that contribute to the effects observed. Alterations in the media could also bring about conformational changes in CMCase leading to increased rates of reaction and subsequent increases in CMC degradation. Finally, explanations for the observed phenomena based on the direct effect of the compounds tested on the cellulosome complex, its domains, and/or its component enzymes are proposed. PMID- 15530017 TI - Discrimination between Candida albicans and Candida dubliniensis isolated from HIV-positive patients by using commercial method in comparison with PCR assay. AB - Nineteen clinical isolates of Candida albicans and C. dubliniensis were isolated from patients (majority of them HIV-positive) in Slovakia, Brazil, Thailand and Japan. Species discrimination was performed by using growth on CHROMagar Candida, commercial biochemical set API 20C AUX, germ-tube test in human serum, growth at 42 and 45 degrees C on Sabouraud-dextrose agar as well as on CHROMagar Candida, assimilation of D-xylose and methyl alpha-D-glucoside by glass-tube test, and production of chlamydospores. These tests were completed by PCR using Cd-oligo2/F and Cd-oligo2/R primer pair specific for C. dubliniensis. Six clinical isolates were confirmed to be C. dubliniensis, remaining 13 strains were determined as C. albicans. The use of conventional method showed that the determination is markedly influenced by personal evaluation suggesting the necessity of using the combination of many tests to obtain correct results comparing with accurate and rapid PCR assay. For discrimination between C. albicans and C. dubliniensis we recommend the combination of primo-cultivation on CHROMagar, followed by germ tube test and PCR. PMID- 15530018 TI - Secreted aspartate proteinases, a virulence factor of Candida spp.: occurrence among clinical isolates. AB - Production of secreted aspartate proteinases was determined in a set of 646 isolates of Candida and non-Candida yeast species collected from 465 patients of the University Hospital in Olomouc (Czechia) in the period 1995-2002, and Candida samples obtained from 64 healthy volunteers using solid media developed for this purpose. Using random amplified polymorphic DNA analysis (RAPD) 79 Candida isolates from blood were analyzed to show potential relationships between clustering of the fingerprints and extracellular proteolytic activity of these strains. C. albicans, C. tropicalis and C. parapsilosis possess always proteolytic activity while non-Candida species did not display any proteolysis. A tight relationship between fingerprints and extracellular proteolysis in the Candida isolates was not shown. A remarkable consistency between fingerprint clusters and proteolysis occurred in a subset of C. parapsilosis samples. Suboptimal pH of the growth medium was shown to facilitate the investigation of potential co-incidence of genotypic and phenotypic traits. PMID- 15530019 TI - AFM observation of band-like cellulose assemblies produced by Acetobacter xylinum. AB - We succeeded in estimating the thickness of band-like cellulose assemblies by combined use of atomic force microscopy (AFM) and transmission electron microscopy. The thickness of "dense" band-like cellulose assemblies was estimated at 20-30 nm from their AFM height profiles, which was several times greater than that of "coarse" band-like and ribbonlike cellulose assemblies. On the basis of these results, the folded- chain model previously proposed was discussed, and a different organization of TC subunits was suggested for the "dense" band-like cellulose assembly. PMID- 15530020 TI - Report on the use of poly(organophosphazenes) for the design of stimuli responsive vesicles. AB - A novel family of amphiphilic temperature- and pH-sensitive poly(organophosphazenes) with varying ratios of ethylene oxide, alkyl chains and free acid units was synthesized by living cationic polymerization. Depending on their composition, these poly(organophosphazenes) exhibited lower critical solution temperatures ranging from 32 to 44 degrees C, which were pH-dependent for copolymers bearing carboxylic acid groups. The alkylated copolymers were then anchored into phospholipid bilayers to obtain stimuli-responsive liposomes that released their content upon a change in temperature or pH. Such polymer/vesicle complexes could find practical applications for site-specific and intracellular drug delivery. PMID- 15530021 TI - Adsorption kinetics and rheological interfacial properties of plant proteins at the oil-water interface. AB - Adsorption and rheological properties of plant proteins were determined by means of the dynamic pendant drop technique. The plant protein properties were compared with the interfacial properties of gelatin, which is well-known for its surface active properties and is commonly used in food and health products. The results showed that alpha gliadins (wheat proteins) and pea globulins have the highest surface active properties at the oil-water interface, even higher than gelatin at the same concentration (weight/volume). After a short time of adsorption, alpha gliadin interfacial behavior is characterized by a pronounced viscoelasticity, which was confirmed with time whereas pea protein interfacial behavior became elastic after a long initial adsorption period. Finally, the behavior of gelatin is very close to the alpha gliadin behavior for the short initial adsorption period, whereas it looks like the behavior of legume seed proteins for longer times of the adsorption kinetics. This study emphasizes the importance of the choice of the proteins and the emulsification time in the encapsulation process, according to the interfacial behavior. PMID- 15530022 TI - Effects on interfacial properties and cell adhesion of surface modification by pectic hairy regions. AB - Polystyrene Petri dishes, aminated by a plasma deposition process, were surface modified by the covalent linking of two different enzymatically modified hairy regions (HRs) from pectin containing, for example, rhamnogalacturonan-I and xylogalacturonan structural elements. The two polysaccharide preparations share the same structural elements of apple pectin, but the relative amounts and lengths of the neutral side chains present differ. Surface analysis by X-ray photoelectron spectroscopy, contact angle measurement, and atomic force microscope (AFM) force-separation curves was used to characterize the effects on surface chemistry and interfacial forces of the surface modification process. Cell adhesion experiments using continuous L-929 fibroblasts and primary aortic smooth muscle cells were performed to evaluate the effect of the polysaccharide nature on cell adhesion. Results show that immobilization of the HR affects the interfacial field of forces and the cell behavior: "equilibrium" contact angles, obtained by a recently introduced vibrational approach, decrease after HR immobilization reaching a value close to 20 degrees . AFM force-separation curves show a more extended (or softer) interface in the case of the HR bearing longer side chains. Accordingly, depending on the HR preparation, cells shifted from spread morphology and adhesion behavior quantitatively comparable to that observed on conventional tissue culture polystyrene to rounded morphology and significantly lower adhesion. These data show that engineering of plant pectins can be a valuable tool to prepare novel and finely tuned polysaccharides having different chemico-physical and biological properties, to be used in the surface modification of medical devices and materials. PMID- 15530023 TI - Secondary structures and conformational changes in flagelliform, cylindrical, major, and minor ampullate silk proteins. Temperature and concentration effects. AB - Orb weaver spiders use exceptionally complex spinning processes to transform soluble silk proteins into solid fibers with specific functions and mechanical properties. In this study, to understand the nature of this transformation we investigated the structural changes of the soluble silk proteins from the major ampullate gland (web radial threads and spider safety line); flagelliform gland (web sticky spiral threads); minor ampullate gland (web auxiliary spiral threads); and cylindrical gland (egg sac silk). Using circular dichroism, we elucidated (i) the different structures and folds for the various silk proteins; (ii) irreversible temperature-induced transitions of the various silk structures toward beta-sheet-rich final states; and (iii) the role of protein concentration in silk storage and transport. We discuss the implication of these results in the spinning process and a possible mechanism for temperature-induced beta-sheet formation. PMID- 15530024 TI - Performance of butyrylcellulose membranes for benzene/cyclohexane mixtures containing a low benzene concentration by pervaporation. AB - Butyrylcellulose (BuCell) with different degrees of butyrylation was synthesized as a membrane material for the separation of benzene/cyclohexane (Bz/Chx) mixtures. A BuCell membrane with a degree of butyrylation of 2.3 showed high benzene/cyclohexane selectivity for Bz/Chx mixtures by pervaporation. Both the permeation rate and the benzene/cyclohexane selectivity of the BuCell membrane increased with increasing benzene concentration in the feed mixture. The increase in the permeation rate resulted from an increase in the swelling of the membrane, and the increase in the benzene/cyclohexane selectivity can be attributed to an increase in the diffusion selectivity. With increasing degree of butyrylation of BuCell, the permeation rate increased; on the other hand, the benzene/cyclohexane selectivity decreased slightly. This result can qualitatively be explained by the degree of swelling, the density, and the contact angle of the BuCell membranes. The permeation and separation mechanism of Bz/Chx mixtures through BuCell membranes by pervaporation is discussed on the basis of the solution-diffusion model, which is typically applied for permeation through dense, nonporous membranes. PMID- 15530025 TI - Evaluation of hyaluronan from different sources: Streptococcus zooepidemicus, rooster comb, bovine vitreous, and human umbilical cord. AB - Sodium hyaluronate (HA) is widely distributed in extracellular matrixes and can play a role in orchestrating cell function. Consequently, many investigators have looked at the effect of exogenous HA on cell behavior in vitro. HA can be isolated from several sources (e.g., bacterial, rooster comb, umbilical cord) and therefore can possess diverse impurities. This current study compares the measured impurities and the differences in biological activity between HA preparations from these sources. It was demonstrated that nucleic acid and protein content was highest in human umbilical cord and bovine vitreous HA and was low in bacterial and rooster comb HA. Macrophages exposed to human umbilical cord HA produced significantly higher amounts of TNF-alpha relative to control or bacterial-derived HA. These results indicate that the source of HA should be considered due to differences in the amounts and types of contaminants that could lead to widely different behaviors in vitro and in vivo. PMID- 15530026 TI - Polyion complex micelles of pDNA with acetal-poly(ethylene glycol)-poly(2 (dimethylamino)ethyl methacrylate) block copolymer as the gene carrier system: physicochemical properties of micelles relevant to gene transfection efficacy. AB - An acetal-poly(ethylene glycol)-poly(2-(dimethylamino)ethyl methacrylate) (acetal PEG-PAMA) block copolymer spontaneously associated with plasmid DNA (pDNA) to form water-soluble complexes (polyion complex micelle: PIC micelle) in aqueous solution. Physicochemical characteristics and transfection efficiency of the PIC micelles thus prepared were studied here, focusing on the residual molar mixing ratio (N/P ratio) of AMA units in acetal-PEG-PAMA to the phosphate units in pDNA. With the N/P ratio increasing to unity, acetal-PEG-PAMA cooperatively formed complex micelles with pDNA through electrostatic interaction, allowing pDNA to condense effectively. Dynamic light scattering measurements revealed that the PIC micelle at N/P > or = 3 had a constant size of approximately 90-100 nm. Eventually, acetal-PEG-PAMA/pDNA micelles underwent no precipitation even after long-term storage for more than 1 month at all N/P ratios. The PIC micelles were stable even in the presence of excess polyanions, poly(vinyl sulfate), in contrast to polyplexes based on the PAMA homopolymer, yet this stabilization effect was highly dependent on the N/P ratio to reach a plateau at N/P = 3-4. This character may be attributed to the increased hydrophobicity in the vicinity of the complexed pDNA. Furthermore, the pDNA in the micelle was adequately protected from DNase I attack. The transfection ability of the PIC micelles toward 293 cells was remarkably enhanced with an increasing N/P ratio as high as 25. The zeta-potential of the micelles with a high N/P ratio was an appreciably large positive value, suggesting a noncooperative micelle formation. This deviated micellar composition with an excess cationic nature as well as the presence of free acetal-PEG-PAMA may play a substantial role in the enhanced transfection efficiency of the PIC micelle system in the high N/P ratio (approximately 25) region. PMID- 15530027 TI - Water structures of differing order and mobility in aqueous solutions of schizophyllan, a triple-helical polysaccharide as revealed by dielectric dispersion measurements. AB - Dielectric dispersion measurements were made on aqueous solutions of a triple helical polysaccharide schizophyllan over a wide concentration range 10-50 wt % at -45 to +30 degrees C. In the solution state, three different water structures with the different relaxation times tau were found, namely, bound water (taul), structured water (taus), and loosely structured water (tauls) in addition to free water (tauP). Structured water is less mobile and loosely structured water is nearly as mobile as free water, but bound water with taul is much less mobile, thus taul >> taus >> tauls greater, similar tauP. The order-disorder transition accompanies the conversion between structured water and loosely structured water. However, the species with taus remains even in the disordered state and constitutes part of bound water in the entire temperature range. In the frozen state, in addition to bulk water formed by partial melting, two mobile species existed, which were assigned to liquidlike bound water and found to be a continuation of bound water in the solution state. These relaxation time data are discussed in connection with the entropy levels of the four structures deduced from heat capacity data (cf. Yoshiba, K.; et al. Biomacromolecules 2003, 4, 1348 1356). PMID- 15530028 TI - Conformational changes of peptides at solid/liquid interfaces: a Monte Carlo study. AB - Monte Carlo simulations were performed to study the conformational changes of negatively charged model peptides dissolved in water adsorbed onto charged surfaces. 8-, 16-, and 20-residues peptides were used, each of them consisted of repeating diblock units of aspartic acid (ASP, polar amino acid) and isoleucine (ILE, nonpolar amino acid) residues. We found that a water patch was retained at the charged surface, separating the peptide from it. We believed that these water molecules were primarily responsible for giving a particular orientation to the peptide at the surface. Water did play a role to some extent in the structural stability of the 8-residues peptide. However, for higher chain lengths (16 residues and 20-residues), the intrinsic hydrogen-bonding network (or intrinsic structural stability) showed a predominant effect over hydrophobic dehydration for the stability of the peptide at the surface. PMID- 15530029 TI - Novel cell adhesive glycosaminoglycan-binding proteins of Japanese encephalitis virus. AB - Glycosaminoglycans (GAGs) are present in the extracellular matrix and/or tissue cell surface and, by binding to specified GAG-binding proteins, control many important cellular functions. Some animal viruses had evolved to use GAGs as part of their strategy to invade host cells. In this study, two putative GAG-binding proteins were identified from the E protein sequence of the live-attenuated strain CH2195LA of Japanese encephalitis virus (JEV): (i) the first GAG-binding region at residues from E-279 to E-297 (279KLTSGHLKCRLKMDKLALK297) and (ii) the second GAG-binding region at residues from E-397 to E-416 (397KAGSTLGKAFFSTTLKGAQR416). Four recombinant proteins with or without these two GAG-binding regions were expressed in Escherichia coli and purified to examine their GAG-binding properties. The first GAG binding region was demonstrated to exhibit a higher affinity in heparin-Sepharase column. Dose-dependent increases of BHK-21 cell binding were also demonstrated by cell binding enzyme-linked immunosorbent assay (ELISA). Immobilized on glass coverslips, the GAG-binding recombinant protein of JEV promoted BHK-21 cell adhesion and proliferation. The present studies demonstrate the recombinant GAG-binding proteins of JEV stimulate cell adhesive and proliferation with a potential for applications in tissue engineering. PMID- 15530030 TI - Structural stability of wild type and mutated alpha-keratin fragments: molecular dynamics and free energy calculations. AB - The objective of this study is to investigate the influence of point mutations on the structural stability of coiled coil fragments of the human hair intermediate filament by molecular dynamics simulations and free energy calculations. Mutations in the helix termination motif of human hair keratin gene hHb6 seem to be connected to the hereditary hair dystrophy Monilethrix. The most common mutations reported are Glu413Lys and Glu413Asp, located at the C-terminal end of the coiled coil 2B rod domain of the IF. According to our simulations, significant conformational changes of the side chains at the mutation and neighboring sites occur due to the Glu413Lys mutation. Furthermore, the differences in electrostatic interactions cause a large change in free energy during transformation of Glu413 to Lys calculated by the thermodynamic integration approach. It is speculated that the structural rearrangement necessary to adapt the interactions in the mutated coiled coil leads to changes in the IF assembly or its stability. The second mutation, Glu413Asp, only leads to a small value of the calculated free energy difference that is within the error limits of the simulations. Thus, it has to be concluded that this mutation does not affect the coiled coil stability. PMID- 15530031 TI - Comprehensive conformational study of key interactions involved in zearalenone complexation with beta-D-glucans. AB - The beta-D-glucans from the cell wall of Saccharomyces cerevisiae have shown in vitro affinity for zearalenone. For this reason, their utilization as dietary adsorbent, to reduce the bioavailability of zearalenone, is of practical interest. Our study used powerful devices to elucidate the spatial conformation and molecular sites of interaction between ZEN and beta-D-glucans. In this respect, 1H NMR spectroscopy implicated the hydroxyl groups of the phenol moiety of zearalenone in the complexation by laminarin, a pure beta-(1,3)-D-glucan. X ray diffraction determined that laminarin displays the conformation of a single helix with six beta-D-glucopyranose residues per turn. At this stage, molecular modeling was useful to locate the interaction sites and to propose highly probable complexes of zearalenone with laminarin fragment. Interestingly, the beta-(1,3)-D-glucan chain favors a very stable intra-helical association with zearalenone, nicely stabilized by beta-(1,6)-D-glucans side chains. Both hydrogen bonds and van der Waals interactions were precisely identified in the complex and could thus be proposed as driving interactions to monitor the association between the two molecules. PMID- 15530032 TI - Enzymatic degradation processes of lamellar crystals in thin films for poly[(R)-3 hydroxybutyric acid] and its copolymers revealed by real-time atomic force microscopy. AB - Enzymatic degradation processes of flat-on lamellar crystals in melt-crystallized thin films of poly[(R)-3-hydroxybutyric acid] (P(3HB)) and its copolymers were characterized by real-time atomic force microscopy (AFM) in a phosphate buffer solution containing PHB depolymerase from Ralstonia pickettii T1. Fiberlike crystals with regular intervals were generated along the crystallographic a axis at the end of lamellar crystals during the enzymatic degradation. The morphologies and sizes of the fiberlike crystals were markedly dependent on the compositions of comonomer units in the polyesters. Length, width, interval, and thickness of the fiberlike crystals after the enzymatic degradation for 2 h were measured by AFM, and the dimensions were related to the solid-state structures of P(3HB) and its copolymers. The width and thickness decreased at the tip of fiberlike crystals, indicating that the enzymatic degradation of crystals takes place not only along the a axis but also along the b and c axes. These results from AFM measurement were compared with the data on crystal size by wide-angle X ray diffraction, and on lamellar thickness and long period by small-angle X-ray scattering. In addition, the enzymatic erosion rate of flat-on lamellar crystals along the a axis was measured from real-time AFM height images. A schematic glacier model for the enzymatic degradation of flat-on lamellar crystals of P(3HB) by PHB depolymerase has been proposed on the basis of the AFM observations. PMID- 15530033 TI - Immobilization and stabilization of recombinant multimeric uridine and purine nucleoside phosphorylases from Bacillus subtilis. AB - We selected the PnpI/PupG (PNP) with specificity for ribo- and deoxyriboguanosine and ribo- and deoxyriboinosine and the Up/Pdp (UP) with specificity for uridine, thymidine, and deoxyuridine from the purine and pyrimidine salvage pathway of the Gram-positive bacterium Bacillus subtilis. Then, an extensive study of the UP (uridine phosphorylase) and PNP (purine nucleoside phosphorylase) immobilization and stabilization was carried out: optimal UP preparation was achieved by immobilization onto Sepabeads coated with poly(ethyleneimine) and finally cross linked with aldehyde dextran (UP-Sep-PEI-Dx); optimal immobilized PNP was prepared onto glyoxyl-agarose. Both derivatives were highly stable and active even under drastic experimental conditions (pH 10, 45 degrees C) unlike the free enzymes which were promptly inactivated. The derivatives prepared were successfully used in the synthesis of 2'-deoxyguanosine by enzymatic transglycosylation in aqueous solution between 2'-deoxyuridine and guanine. PMID- 15530034 TI - Interactions of two amphiphilic penicillins with myoglobin in aqueous buffered solutions: a thermodynamic and spectroscopy study. AB - The interactions and complexation process of the amphiphilic penicillins sodium cloxacillin and sodium dicloxacillin with horse myoglobin in aqueous buffered solutions of pH 4.5 and 7.4 have been examined by equilibrium dialysis, zeta potential, isothermal titration calorimetry (ITC) and UV-Vis absorbance techniques. A more opened structure of the protein molecules is detected as a consequence of the reduction of pH from 7.4 to 4.5. Binding isotherms and derived Hill coefficients reflect a cooperative binding behavior. Gibbs energies of binding per mole of drug were obtained from equilibrium dialysis data and compared with those derived from the zeta potential taking into account cooperativity. DeltaGads degrees values so obtained are large and negative at low concentrations where binding to the "high-energy" sites occurs and decreases with the drug concentration. The enthalpies of binding have been obtained from ITC and are small and exothermic so that the Gibbs energies of binding are dominated by large increases in entropy consistent with hydrophobic interactions. Other thermodynamic quantities of the binding mechanism, that is, entropy, DeltaSITCi, Gibbs energy, DeltaGITCi, the binding constant, KITCi, and the number of binding sites, ni, were also obtained, confirming the above results. From ITC data and following a theoretical model, the number of bound and free penicillin molecules was calculated, being higher at pH 4.5 than at pH 7.4. The binding of penicillin causes a conformational transition on protein structure as a consequence of the resulting intramolecular repulsion between the penicillin molecules bound to the protein. Thermodynamic quantites (the Gibbs energy of the transition in water, DeltaGw degrees , and in a hydrophobic environment, DeltaGhc degrees) of the denaturation process were calculated, indicating that at pH 4.5 some of the histidine residues are protonated, becoming accessible to solvent and giving rise to a more opened protein structure. PMID- 15530035 TI - Fabrication and optimization of methylphenoxy substituted polyphosphazene nanofibers for biomedical applications. AB - Electrospinning has developed as a unique and versatile process to fabricate ultrathin fibers in the form of nonwoven meshes or as oriented arrays from a variety of polymers. The very small dimension of these fibers can generate a high surface area, which makes them potential candidates for various biomedical and industrial applications. The objective of the present study was to develop nanofibers from polyphosphazenes, a class of inorganic-organic polymers known for high biocompatibility, high-temperature stability, and low-temperature flexibility. Specifically, we evaluated the feasibility of developing bead-free nonwoven nanofiber mesh from poly[bis(p-methylphenoxy)phosphazene] (PNmPh) by electrospinning. The effect of process parameters such as nature of solvent, concentration of the polymer solution, effect of needle diameter, and applied potential on the diameter and morphology (beaded or bead-free) of resulting nanofibers were investigated. It was found that solution of PNmPh in chloroform at a concentration range of 7% (wt/v) to 9% (wt/v) can be readily electrospun to form bead-free fibers at room temperature. The mean diameter of the fibers obtained under optimized spinning condition was found to be approximately 1.2 microm. The bead-free, cylindrical nanofibers formed under the optimized condition showed a slightly irregular surface topography with indentations of a few nanometer scale. Further, the electrospun nanofiber mats supported the adhesion of bovine coronary artery endothelial cells (BCAEC) as well as promoted the adhesion and proliferation of osteoblast like MC3T3-E1 cells. PMID- 15530036 TI - Nanoscopic cationic methacrylate star homopolymers: synthesis by group transfer polymerization, characterization and evaluation as transfection reagents. AB - Seven star polymers with degrees of polymerization (DPs) of the arms from 10 to 100 and dimensions in the nanometer range were prepared using sequential group transfer polymerization of 2-(dimethylamino)ethyl methacrylate (DMAEMA, hydrophilic positively ionizable monomer) and ethylene glycol dimethacrylate (hydrophobic neutral cross-linker). The polymers were characterized in tetrahydrofuran by gel permeation chromatography and static light scattering to determine the molecular weights and the weight-average number of arms for each sample. The number of arms of the star polymers varied from 20 to 72. Aqueous solutions of the star polymers were studied by turbidimetry, hydrogen ion titration, and dynamic light scattering to determine their cloud points, pKs, and hydrodynamic diameters. The cloud points of the larger star polymers, with arm DP 30-100, were found to be 29-34 degrees C, almost independent of the DP of the arms. Similarly, the pKs of all star polymers were calculated to range between 6.7 and 7.0, again independent of the arm DP. In contrast, the hydrodynamic diameters of the star polymers strongly depended on the DP of the arms. In particular, by increasing the DP of the arms from 20 to 100, the hydrodynamic diameters in water increased from 7 to 31 nm. All star polymers were evaluated for their ability to transfect human cervical HeLa cancer cells with the modified plasmid pRLSV40 with the enhanced green fluorescent protein as the reporter gene. Our results showed that as the DP of the arms of the DMAEMA star homopolymers increased from 10 to 100, the overall transfection efficiency decreased, with the star polymer with DP of the arms of 10 emerging as the best transfection reagent. Systematic variation of the amounts of star polymer and plasmid DNA used in the transfections led to an optimization of the performance of this star polymer, yielding overall transfection efficiencies of 15%, comparable to the optimum overall transfection efficiency of the commercially available transfection reagent SuperFect of 13%. PMID- 15530037 TI - Engineering lipobeads: properties of the hydrogel core and the lipid bilayer shell. AB - We previously reported on a novel system termed Lipobead that consists of hydrogel beads encased within an anchored lipid bilayer. The hydrogel particles are formed by inverse suspension polymerization of dimethylacrylamide with N,N' ethylenebis(acrylamide). During the polymerization stage, the water in oil emulsion is interfacially stabilized by small molecule surfactants as well as a small percentage of lipid functionalized with a vinyl group. The functionalized lipid becomes tethered to the bead surface and promotes the assembly of a lipid bilayer on the surface of the hydrogel beads. The presence of the functionalized lipid during polymerization dramatically alters the yield, average size, and size distribution of beads produced. This paper examines the effect of various chemical and physical processing parameters on the average size and size distribution of beads produced when lipid is a component of the surfactant mixture. Relationships between the processing parameters, average bead size, and size distribution were established. Macroscopic properties of the lipid bilayers of Lipobeads were also evaluated including phase transition temperature as well as permeability to the small polar molecule, adenosine triphosphate. It was established that the presence of functionalized lipid improves the organization of the bilayer on the Lipobead surface. PMID- 15530038 TI - Assessment of in vitro bioactivity of hyaluronic acid and sulfated hyaluronic acid functionalized electroactive polymer. AB - Electrically conductive polypyrrole (PPY) was surface functionalized with hyaluronic acid (HA) and sulfated hyaluronic acid (SHA) to improve its surface biocompatibility. The immobilization of HA on the PPY film was facilitated by the use of a cross-linker having the appropriate functional groups. The biological activity of the HA functionalized PPY film was assessed by means of an in vitro PC12 cell culture. The cell attachment on different substrates was studied and determined by bicinchoninic acid protein analysis. Cell attachment on the HA functionalized PPY film surface was significantly enhanced in the presence of nerve growth factor. The SHA functionalized PPY film was obtained by the sulfonation of the immobilized HA using pyridinesulfonate. The retention of the biological activity of the immobilized HA after sulfonation was evaluated by the in vitro assessment of the plasma recalcification time (PRT) and platelet adhesion on the substrate. The PRT observed from the SHA functionalized PPY film was significantly prolonged compared with the HA functionalized PPY. Some reduction of platelet adhesion was observed for the SHA functionalized PPY film, compared with that of the HA functionalized PPY film. PMID- 15530039 TI - Study of protein conformation and orientation in silkworm and spider silk fibers using Raman microspectroscopy. AB - Raman microspectroscopy has been used for the first time to determine quantitatively the orientation of the beta-sheets in silk monofilaments from Bombyx mori and Samia cynthia ricini silkworms, and from the spider Nephila edulis. It is shown that, for systems with uniaxial symmetry such as silk, it is possible to determine the order parameters P2 and P4 of the orientation distribution function from intensity ratios of polarized Raman spectra. The equations allowing the calculation of P2 and P4 using polarized Raman microspectroscopy for a vibration with a cylindrical Raman tensor were first derived and then applied to the amide I band that is mostly due to the C=O stretching vibration of the peptide groups. The shape of the Raman tensor for the amide I vibration of the beta-sheets was determined from an isotropic film of Bombyx mori silk treated with methanol. For both the Bombyx mori and Samia cynthia ricini fibroin fibers, the values of P2 and P4 obtained are equal to 0.36 +/- 0.03 and 0.19 +/- 0.02, respectively, even though the two types of silkworm fibroins strongly differ in their primary sequences. For the Nephila edulis dragline silk, values of P2 and P4 of -0.32 +/- 0.02 and 0.13 +/- 0.02 were obtained, respectively. These results clearly indicate that the carbonyl groups are highly oriented perpendicular to the fiber axis and that the beta sheets are oriented parallel to the fiber axis, in agreement with previous X-ray and NMR results. The most probable distribution of orientation was also calculated from the values of P2 and P4 using the information entropy theory. For the three types of silk, the beta-sheets are highly oriented parallel to the fiber axis. The orientation distributions of the beta-sheets are nearly Gaussian functions with a width of 32 degrees and 40 degrees for the silkworm fibroins and the spider dragline silk, respectively. In addition to these results, the comparison of the Raman spectra recorded for the different silk samples and the polarization dependence of several bands has allowed to clarify some important band assignments. PMID- 15530040 TI - Novel biodegradable aliphatic poly(butylene succinate-co-cyclic carbonate)s bearing functionalizable carbonate building blocks: II. Enzymatic biodegradation and in vitro biocompatibility assay. AB - In a previous study, we have reported chemical synthesis of novel aliphatic poly(butylene succinate-co-cyclic carbonate) P(BS-co-CC)s bearing various functionalizable carbonate building blocks, and this work will continue to present our new studies on their enzymatic degradation and in vitro cell biocompatibility assay. First, enzymatic degradation of the novel P(BS-co-CC) film samples was investigated with two enzymes of lipase B Candida Antartic (Novozyme 435) and lipase Porcine Pancreas PPL, and it was revealed that copolymerizing linear poly(butylene succinate) PBS with a functionalizable carbonate building block could remarkably accelerate the enzymatic degradation of a synthesized product P(BS-co-CC), and its biodegradation behavior was found to strongly depend on the overall impacts of several important factors as the cyclic carbonate (CC) comonomer structure and molar content, molar mass, thermal characteristics, morphology, the enzyme-substrate specificity, and so forth. Further, the biodegraded residual film samples and water-soluble enzymatic degradation products were allowed to be analyzed by means of proton nuclear magnetic resonance (1H NMR), gel permeation chromatograph (GPC), differential scanning calorimeter (DSC), attenuated total reflection FTIR (ATR-FTIR), scanning electron microscope (SEM), and liquid chromatograph-mass spectrometry (LC-MS). On the experimental evidences, an exo-type mechanism of enzymatic chain hydrolysis preferentially occurring in the noncrystalline domains was suggested for the synthesized new P(BS-co-CC) film samples. With regard to their cell biocompatibilities, an assay with NIH 3T3 mouse fibroblast cell was conducted using the novel synthesized P(BS-co-CC) films as substrates with respect to the cell adhesion and proliferation, and these new biodegradable P(BS-co-CC) samples were found to exhibit as low cell toxicity as the PLLA control, particularly the two samples of poly(butylene succinate-co-18.7 mol % dimethyl trimethylene carbonate) P(BS-co-18.7 mol % DMTMC) and poly(butylene succinate-co-21.9 mol % 5 benzyloxy trimethylene carbonate) P(BS-co-21.9 mol % BTMC) were interestingly found to show much better cell biocompatibilities than the PLLA reference. PMID- 15530041 TI - In vitro targeting of synthesized antibody-conjugated dendrimer nanoparticles. AB - This study reports the synthesis and in vitro biological properties of dendrimer antibody conjugates. The polyamidoamine dendrimer platform was conjugated to fluorescein isothiocyanate as a means to analyze cell binding and internalization. Two different antibodies, 60bca and J591, which bind to CD14 and prostate-specific membrane antigen (PSMA), respectively, were used as model targeting molecules. The binding of the antibody-conjugated dendrimers to antigen expressing cells was evaluated by flow cytometry, confocal microscopy, and a new two-photon-based optical fiber fluorescence detection system. The conjugates specifically bound to the antigen-expressing cells in a time- and dose-dependent fashion, with affinity similar to that of the free antibody. Confocal microscopic analysis suggested at least some cellular internalization of the dendrimer conjugate. Dendrimer-antibody conjugates are a suitable platform for targeted molecule delivery into antigen-expressing cells. PMID- 15530042 TI - Electron beam-induced graft polymerization of acrylic acid and immobilization of arginine-glycine-aspartic acid-containing peptide onto nanopatterned polycaprolactone. AB - Electron beam- (EB-) induced graft polymerization of acrylic acid and the subsequent immobilization of arginine-glycine-aspartic acid (RGD) peptide onto nanopatterned polycaprolactone with parallel grooves is reported. A high concentration of carboxylic groups was introduced onto the polymer substrate by EB-induced polymerization of acrylic acid. In the coupling of the RGD peptide to the carboxylated polymer surface, a three-step peptide immobilization process was used. This process included the activation of surface carboxylic acid into an active ester intermediate by use of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS), the introduction of disulfide groups by use of 2-(2-pyridinyldithio)ethanamine hydrochloride (PDEA), and final immobilization of the peptide via a thiol-disulfide exchange reaction. The extent of coupling was measured by UV spectroscopy. A preliminary study of the in vitro behavior of keratinocytes (NCTC 2544) cultured on the acrylic acid-grafted and RGD peptide-coupled surface showed that most cells grown on the coupled samples had a spread-rounded appearance, while the majority of cells tended to be elongated along the grooves on uncoupled substrates. PMID- 15530043 TI - "Green" nanocomposites from cellulose acetate bioplastic and clay: effect of eco friendly triethyl citrate plasticizer. AB - "Green" nanocomposites have been successfully fabricated from cellulose acetate (CA) powder, eco-friendly triethyl citrate (TEC) plasticizer and organically modified clay. The effect of the amount of plasticizer varying from 15 to 40 wt % on the performance of the nanocomposites has been evaluated. The morphologies of these nanocomposites were evaluated through X-ray diffraction (XRD), atomic force microscopy (AFM), and transmission electron microscopy (TEM) studies. The mechanical properties of nanocomposites are correlated with the XRD and TEM observations. Cellulosic plastic-based nanocomposites with 20 wt % TEC plasticizer and 5 wt % organoclay showed better intercalation and an exfoliated structure than the counterpart having 30/40 wt % plasticizers. The tensile strength, modulus and thermal stability of cellulosic plastic reinforced with organoclay showed a decreasing trend with an increase of plasticizer content from 20 to 40 wt %. The nano-reinforcement at the lower volume fractions (phi < or = 0.02) reduced the water vapor permeability of cellulosic plastic by 2 times and the relative permeability better fits with larger platelet aspect ratios (alpha = 150). PMID- 15530044 TI - Comparison between denaturant- and temperature-induced unfolding pathways of protein: a lattice Monte Carlo simulation. AB - Denaturant-induced unfolding of protein is simulated by using a Monte Carlo simulation with a lattice model for protein and denaturant. Following the binding theory for denaturant-induced unfolding, the denaturant molecules are modeled to interact with protein by nearest-neighbor interactions. By analyzing the conformational states on the unfolding pathway of protein, the denaturant-induced unfolding pathway is compared with the temperature-induced unfolding pathway under the same condition; that is, the free energies of unfolding under two different pathways are equal. The two unfoldings show markedly different conformational distributions in unfolded states. From the calculation of the free energy of protein as a function of the number fraction (Q0) of native contacts relative to the total number of contacts, it is found that the free energy of the largely unfolded state corresponding to low Q0 (0.1 < Q0 < 0.5) under temperature induced unfolding is lower than that under denaturant-induced unfolding, whereas the free energy of the unfolded state close to the native state (Q0 > 0.5) is lower in denaturant-induced unfolding than in temperature-induced unfolding. A comparison of two unfolding pathways reveals that the denaturant-induced unfolding shows a wider conformational distribution than the temperature-induced unfolding, while the temperature-induced unfolding shows a more compact unfolded state than the denaturant-induced unfolding especially in the low Q0 region (0.1 < Q0 < 0.5). PMID- 15530045 TI - Molecular chain orientation of DNA films induced by both the magnetic field and the interfacial effect. AB - DNA films showing highly homogeneous orientation of molecular chains were successfully prepared by drying a semidiluted solution in a horizontal magnetic field. Most of the molecular chain elements in the obtained film were found to be one-dimensionally oriented, as shown by X-ray diffraction, polarization microscopy, and linear dichroism spectroscopy. Because a DNA chain is theoretically expected to orientate only in divergent directions perpendicular to a magnetic field, this result suggests that the DNA chains were aligned not only by a magnetic field but also by the interfacial effect that induced the chains to fit along the air-liquid interface. The descent speed of an air-liquid interface by evaporation was faster than the estimated diffusion rate of DNA, suggesting an emergence of a concentrated layer near the surface. As proved by polarization microscopy, this emergence led to the transitional formation of a nematic-like liquid crystalline phase, which resulted in a DNA film with good chain alignment and unitary orientation. This mechanism underlying chain alignment was supported by molecular weight dependency, in which higher molecular weight DNA is more likely to evince chain alignment that exhibits a higher degree of birefringence. Low molecular weight components have such high thermal motility that it would be difficult to fit them along the air-liquid interface in the early stage of drying. For chain alignment, it was preferable to use an initial concentration of DNA lower than a critical concentration for liquid crystal formation so that the possible diffusion and assembly in a diluted solution would be essential for chain alignment. The DNA film exhibited obvious linear dichroism, indicating the potential for further applications. PMID- 15530046 TI - Control of nanobiointerfaces generated from well-defined biomimetic polymer brushes for protein and cell manipulations. AB - To better understand protein/material and cell/material interactions at the submolecular level, well-defined polymer brushes consisting of poly(2 methacryloyloxyethyl phosphorylcholine) (PMPC) on silicon wafers were prepared by atom transfer radical polymerization (ATRP). Silicon wafers were treated with 3 (2-bromoisobutyryl)propyl dimethylchlorosilane (BDCS) to form a monolayer that acts as initiators for ATRP. Silicon-supported BDCS monolayers were soaked in a methanol/water mixture solution containing Cu(I)Br, bipyridine, and a sacrificial initiator. After MPC was added to the solution, ATRP was carried out for 18 h. The molecular weight and thickness of the PMPC brush layer on the silicon surface increased with an increase in the polymerization time. The dense polymer brushes were obtained by the "grafting from" system. By selective decomposition of the BDCS monolayer by UV light-irradiation, the PMPC brush region and the sizes were well controlled, resulting in fabricating micropatterns of the PMPC brushes. When the thickness of the PMPC brush layer was greater than 5.5 +/- 1.0 nm (3 h polymerization), serum protein adsorption and fibroblast adhesion were effectively reduced, i.e., proteins and cells could recognize such thin polymer brushes on the surface. In addition, the density of the adherent cells on the patterned PMPC brush surface could be controlled by changing the size of the pattern. PMID- 15530047 TI - Light-activated immobilization of biomolecules to agarose hydrogels for controlled cellular response. AB - We describe a new method of synthesizing photolabile hydrogel materials for convenient photoimmobilization of biomolecules on surfaces or in 3-D matrixes. Dissolved agarose was modified with photolabile S-(2-nitrobenzyl)cysteine (S-NBC) via 1,1'-carbonyldiimidazole (CDI) activation of primary hydroxyl groups. S-NBC modified agarose remained soluble and gelable with up to 5% S-NBC substitution, yet gelation was slower and the elastic modulus of the resulting gel was lower than those of unmodified agarose. Irradiating S-NBC-grafted agarose resulted in the loss of the protecting 2-nitrobenzyl groups, thereby exposing free sulfhydryl groups for biomolecular coupling. When appropriately activated with sulfhydryl reactive groups, either peptides or proteins were effectively immobilized to the photoirradiated hydrogel matrixes, with the irradiation energy dose (i.e., irradiation time) used to control the amount of biomolecule immobilization. When the GRGDS peptide was immobilized on agarose, it was shown to be cell-adhesive and to promote neurite outgrowth from primary, embryonic chick dorsal root ganglion neurons. The immobilized GRGDS surface ligand concentration affected the cellular response: neurite length and density increased with GRGDS surface concentration at low adhesion ligand concentration and then plateaued at higher GRGDS concentration. Grafting 2-nitrobenzyl-protected compounds to hydrogel materials is useful for creating new photolabile hydrogel substrates for light activated functional group generation and biomolecular immobilization. PMID- 15530048 TI - DNA-induced aggregation of zwitterionic oligolamellar liposome. AB - Liposome consisting of a single zwitterionic lipid as the potential vector for gene therapy has been reported recently; however, whether polyanionic DNA can bind directly with zwitterionic lipid without the aid of multivalent salt still remains unresolved. In this study, we reveal the aggregation of zwitterionic oligolamellar liposomes composed of 1,2-di(cis-9-octadecenoyl)-sn-glycero-3 phosphocholine induced by DNA without the presence of multivalent salt. Our results demonstrate that only a small fraction (<10%) of DNA can bind electrostatically with a portion of the liposomes. Such a low degree of binding, however, induces significant aggregation of these oligolamellar liposomes, yielding large multilamellar particles in which the number of hydrophilic/hydrophobic layer stacking becomes sufficiently large to yield multiple diffraction peaks in the small-angle X-ray scattering profile. Addition of monovalent salt such as NaCl tends to disrupt the multilamellar structure. PMID- 15530049 TI - Extruding foams from corn starch acetate and native corn starch. AB - Because of the hydrophilic characteristics of native starch foams and the cost of modifying starch, the uses of starch and modified starch foams are hindered. To decrease hydrophilicity and cost of starch foams, native corn starch was blended with starch acetate and extruded. A twin-screw mixing extruder was used to produce the foams. Native starch content, screw speed, and barrel temperature had significant effects on molecular degradation of starches during extrusion. The melting temperature of extruded starch acetate/native starch foam was higher (216 degrees C) than that for starch acetate (193.4 degrees C). Strong peaks in the X ray diffractograms of extruded starch acetate/native starch foam suggested new crystalline regions were formed. Optimum conditions for high radial expansion ratio, high compressibility, low specific mechanical energy requirement, and low water absorption index were 46.0% native starch content, 163 rpm screw speed, and 148 degrees C barrel temperature. PMID- 15530050 TI - Synthesis of chitosan-stabilized gold nanoparticles in the absence/presence of tripolyphosphate. AB - Gold nanoparticles were prepared by reducing gold salt with a polysaccharide, chitosan, in the absence/presence of tripolyphosphate (TPP). Here, chitosan acted as a reducing/stabilizing agent. The obtained gold nanoparticles were characterized with UV--vis spectroscopy and transmission electron microscopy. The results indicated that the shape and size distribution of gold nanoparticles changed with the molecular weight and concentration of chitosan. More interestingly, the gelation of chitosan upon contacting with polyanion (TPP) can also affect the shape and size distribution of gold nanoparticles. By adding TPP to chitosan solution before the reduction of gold salt, gold nanoparticles have a bimodal size distribution, and at the same time, polygonal gold particles were obtained in addition to spherical gold nanoparticles. PMID- 15530052 TI - In situ gelling of alginate/pluronic solutions for ophthalmic delivery of pilocarpine. AB - We prepared a series of alginate and Pluronic-based solutions as the in situ gelling vehicles for ophthalmic delivery of pilocarpine. The rheological properties, in vitro release as well as in vivo pharmacological response of polymer solutions, including alginate, Pluronic solution, and alginate/Pluronic solution, were evaluated. The optimum concentration of alginate solution for the in situ gel-forming delivery systems was 2% (w/w) and that for Pluronic solution was 14% (w/w). The mixture of 0.1% alginate and 14% Pluronic solutions showed a significant increase in gel strength in the physiological condition; this gel mixture was also found to be free flowing at pH 4.0 and 25 degrees C. Both in vitro release and in vivo pharmacological studies indicated that the alginate/Pluronic solution retained pilocarpine better than the alginate or Pluronic solutions alone. The results demonstrated that the alginate/Pluronic mixture can be used as an in situ gelling vehicle to increase ocular bioavailability. PMID- 15530051 TI - Measurement of normal and anomalous diffusion of dyes within protein structures fabricated via multiphoton excited cross-linking. AB - We demonstrate microscale spatial and chemical control of diffusion within protein matrixes created through the use of nonlinear multiphoton excited photochemistry. The mobility of fluorescent dyes of different mass and composition within controlled cross-linked environments has been measured using two-photon excited fluorescence recovery after photobleaching (FRAP). The diffusion times for several rhodamine and sulforhodamine dyes within these fabricated structures were found to be approximately 3-4 orders of magnitude slower than in free solution. The precise diffusion times can be tuned by varying the laser exposure during the fabrication of the matrix, and the diffusion can be correlated with the mesh size determined by TEM and Flory-Rehner analysis. We find that the hydrophobic Texas Red dyes (sulforhodamines) exhibit diffusion that is highly anomalous, indicative of a strong interaction with the hydrophobic cross-linked protein matrix. These results suggests the use of these cross-linked protein matrixes as ideal model systems in which to systematically study anomalous diffusion. Finally, the diffusion can be tuned within a multilayered protein matrix, and this in conjunction with slow diffusion also suggests the use of these structures in controlled release applications. PMID- 15530053 TI - Computational approach to solvent-free synthesis of ethyl oleate using Candida rugosa and Candida antarctica B Lipases. I. Interfacial activation and substrate (ethanol, oleic acid) adsorption. AB - This paper presents the results of a MM2 study of the adsorption of oleic acid and ethanol/water in the tunnel and active-site models of lipases from Candida rugosa and Candida antarctica B. The role of an interface polar/no polar in the opening of C. rugosa lipase's lid is also addressed, discussed and analyzed at the level of the conformational changes needed to achieve the lipase open form. The adsorption of oleic acid and alcohols considering C. antarctica B, a lipase not interfacially activated, is also presented. In this case, the tunnel is shorter than in case of C. rugosa lipase. Two different pockets can be visualized at the active site-tunnel model of C. antarctica B lipase: one for the acyl group and another for the alcohol. Wrong location of alcohol and oleic acid severely hinders reaction because it hinders the H-transfer to histidine, a key step in the reaction mechanism. Right location of alcohol decreases the possibility of alcohol inhibition. In the case of C. rugosa, no restrictions for ethanol/water location are found. For that lipase, a second adsorption site for oleic acid (outside the tunnel) is presented. This site is the exit tunnel of the ester product when oleic acid is adsorbed in the tunnel. Experimental results of our own that correlate with this study are presented. PMID- 15530054 TI - Enzymatic cross-linking of a phenolic polymer extracted from the marine alga Fucus serratus. AB - We have shown that a phenolic polymer (PP) extracted from Fucus serratus can be cross-linked using a vanadium-dependent bromoperoxidase (BPO). The methanol extracted PP was adsorbed to a quartz crystal sensor and the cross-linking was initiated by the addition of BPO, KBr, and H2O2. The decreased dissipation upon addition of the cross-linking agents, as measured with the quartz crystal microbalance with dissipation monitoring (QCM-D) method, was interpreted as intramolecular cross-links were formed between different phloroglucinol units in the PP. With surface plasmon resonance, it was shown that no desorption occurred from the sensor surface during the cross-linking. UV/vis spectroscopy verified the results achieved with QCM-D that all components, i.e., BPO, KBr, and H2O2, were necessary in order to achieve intramolecular oxidative cross-linking of the polymer. PMID- 15530055 TI - Solution properties of the xyloglucan polymer from Afzelia africana. AB - In this paper we describe the solution properties of a new xyloglucan polysaccharide extracted from the African legume Afzelia africana Se. Pers. The polysaccharide is of high weight-average molecular weight (Mw), but application of the "pressure cell" method enabled a range of Mw fractions to be prepared. Results from the light scattering/intrinsic viscosity measurements on these fractions suggest that like other xyloglucans from tamarind and detarium it occurs in solution as a polymeric coil, with a small amount of excluded volume. Measurement of dilute and semidilute solution rheology suggests that, like these polymers, and the related galactomannan series, it forms viscous solutions at higher concentrations via entanglements. PMID- 15530056 TI - Surface-active and stimuli-responsive polymer--Si(100) hybrids from surface initiated atom transfer radical polymerization for control of cell adhesion. AB - A simple two-step method was developed for the covalent immobilization of atom transfer radical polymerization (ATRP) initiators on the hydrogen-terminated Si(100) (Si-H) surface. Well-defined functional polymer-Si hybrids, consisting of covalently tethered brushes of poly(ethylene glycol) monomethacrylate (PEGMA) polymer, N-isopropylacrylamide (NIPAAm) polymer, and NIPAAm-PEGMA copolymers and block copolymers on Si-H surfaces, were prepared via surface-initiated ATRP. Kinetics study revealed that the chain growth from the silicon surface was consistent with a "controlled" process. Surface cultures of the cell line 3T3 Swiss albino on the hybrids were evaluated. The PEGMA graft-polymerized silicon [Si-g-P(PEGMA)] surface is very effective in preventing cell attachment and growth. At 37 degrees C [above the lower critical solution temperature (LCST, approximately 32 degrees C) of NIPAAm], the seeded cells adhered, spread, and proliferated on the NIPAAm graft polymerized silicon [Si-g-P(NIPAAm)] surface. Below the LCST, the cells detached from the Si-g-P(NIPAAm) surface spontaneously. Incorporation of PEGMA units into the NIPAAm chains of the Si-g-P(NIPAAm) surface via copolymerization resulted in more rapid cell detachment during the temperature transition. The "active" chain ends on the Si-g-P(PEGMA) and Si-g P(NIPAAm) hybrids were also used as the macroinitiators for the synthesis of diblock copolymer brushes. Thus, not only are the hybrids potentially useful as stimuli-responsive adhesion modifiers for cells in silicon-based biomedical microdevices but also the active chain ends on the hybrid surfaces offer opportunities for further surface functionalization and molecular design. PMID- 15530057 TI - Nanoscale self-assembly of multiblock copolymer chains into rods. AB - Rodlike fibrous structures are increasingly found with biomacromolecules, e.g. silks, elastin, and collagen. Here we propose a multiblock copolymer model that can lead to these larger-scale structures. One component consists of highly regular blocks, whereas the blocks of another, incompatible component exhibit a wide range of lengths. Rods readily form when these lengths have a bimodal distribution and a well-defined periodicity along the chain. Orientation in a flowing liquid assists association of rods, as is likely in the spinning of silks, which have comparable repetitive sequences. The model suggests that rodlike structures should be available in similarly designed synthetic polymers. This route would be distinct from existing routes that employ rigid monomers, e.g. aramids. PMID- 15530058 TI - Fibrillar beta-lactoglobulin gels: Part 1. Fibril formation and structure. AB - As a prelude to experimental and theoretical work on the mechanical properties of fibrillar beta-lactoglobulin gels, this paper reports the structural characterization of beta-lactoglobulin fibrils by electron and atomic force microscopy (AFM), infrared and Raman spectroscopy, and powder X-ray diffraction. Aggregates formed by incubation of beta-lactoglobulin in various alcohol-water mixtures at pH 2, and in water-trifluoroethanol (TFE) at pH 7, were found to be wormlike (approximately 7 nm in width and <500 nm in length), with a "string-of beads" appearance. Longer (approximately 7 nm in width, and >1 microm in length), smoother, and seemingly stiffer fibrils formed on heating aqueous beta lactoglobulin solutions at pH 2 and low ionic strength, although there was little evidence for the higher-order structures common in most amyloid-forming systems. Time-lapse AFM also revealed differences in the formation of these two fibril types: thermally induced aggregation occurring more cooperatively, in keeping with a nucleation and growth process. Only short stiff-rods (<20 nm in length) formed on heating beta-lactoglobulin at pH 7, and only complex three-dimensional "amorphous"aggregates in alcohols other than TFE at this pH. Studies of all of the pH 2 fibrils from beta-lactoglobulin, by Raman and infrared spectroscopy confirmed beta-sheet as mediating the aggregation process. Interestingly, however, some evidence for de novo helix formation for the solvent-induced systems was obtained, although it remains to be seen whether this is actually incorporated into the fibril-structure. In contrast to other amyloid systems, X ray powder diffraction provided no evidence for extensive repeating "crystalline" structures for any of the pH 2 beta-lactoglobulin fibrils. In relation to amyloid, the lactoglobulin fibrils bear more resemblance to protofilaments than to higher-order fibril structures, these latter appearing more convincingly for thermally induced insulin fibrils (pH 2) also included in the AFM study. PMID- 15530059 TI - Fibrillar beta-lactoglobulin gels: Part 2. Dynamic mechanical characterization of heat-set systems. AB - Oscillatory shear rheometry (mechanical spectroscopy) has been used to study the heat-set gelation of beta-lactoglobulin at pH 2. Modulus-concentration relationships were obtained by extrapolating cure data to infinite time. In terms of theory, these fail to provide a clear distinction between the fractal description of biopolymer gels and the classical random f-functional polycondensation branching theory (cascade) approach, though the latter is preferred. Critical exponents for the sol-gel transition, derived from these data, are also discussed. Where gel time-concentration results are concerned the fractal model makes no predictions, and the cascade approach in its simplest form must be rejected in favor of a more sophisticated version involving delivery of fibrils by nucleation and growth into the random aggregation process. Over the limited concentration range accessed experimentally, cure data for the different beta-lactoglobulin solutions, reduced to the universal form G'/G'inf versus t/tgel, superimposed well for samples heated both at 80 and 75 degrees C and for different batches of protein. Studies of the frequency responses of the fully cured gels confirm the validity of the gel description given to these materials, and a study of the temperature dependence of the frequency spectrum suggests a fall in the elastic component of the modulus as temperature decreases. This contrasts with what has been found for other heat-set globular protein gels such as those from serum albumin where the gel modulus increases at lower temperatures. The present results are in good agreement with more limited amounts of pH 2 beta-lactoglobulin data published earlier, though some differences arise through a previous neglect of measurement "dead time". PMID- 15530060 TI - Fibrillar beta-lactoglobulin gels: Part 3. Dynamic mechanical characterization of solvent-induced systems. AB - Oscillatory shear rheometry has been used to study the gelation of beta lactoglobulin at ambient in 50% v/v trifluoroethanol (TFE)/pH 7 aqueous buffer and in 50% v/v ethanol (EtOH)/water at pH 2. In contrast to what was found on heating aqueous solutions at pH 2 (Part 2 of this series), a more expected "chemical gelation"-like profile was found with modulus components G' and G' ' crossing over as the gels formed and then with G' ' passing through a maximum. In addition, for the EtOH system, there was a significant modulus increase at long time, suggestive of a more complex two-step aggregation scheme. Modulus concentration relationships were obtained for both systems by extrapolating cure data to infinite time. For the TFE gels, this data was accurately described by classical branching theory, although it could also be approximated by a constant power--law relationship. Only the latter described the modulus--concentration data for the gels in ethanol, but there were problems here of greater frequency dependence of the modulus values and much less certain extrapolation. Gel times for the TFE systems showed higher power laws in the concentration than could be explained by the branching theory in its simplest form being similar, in this respect, to the heat-set systems at pH 2. Such power laws were harder to establish for the EtOH gels as for these there was evidence of gel time divergence close to a critical concentration. Reduced G'/G'inf versus t/tgel data were difficult to interpret for the gels in ethanol, but for the TFE system they were consistent with previous results for the heat-set gels and approximated master curve superposition. The frequency and temperature dependences of the final gel moduli were also studied. In general, the networks induced by alcohols appeared more flexible than those obtained by heating. PMID- 15530061 TI - Study of gelling behavior of poly(vinyl alcohol)-methacrylate for potential utilizations in tissue replacement and drug delivery. AB - The need of innovative, multifunctional biomaterials for the partial or complete tissue replacement is the driving force for the search of improvements of the performances of the available materials and in the formulation of new ones. Addressing the focus to vitreous substitution, we have explored the possibility of using injectable aqueous solutions of poly(vinyl alcohol), PVA, derivatives able to form hydrogels in the ocular cavity upon UV-vis irradiation with visible light. In particular, we describe the features of hydrogels from methacrylate grafted PVA, PVA-MA, in terms of structural characteristics, degradation processes, release of low- and high- molecular weight molecules, and in vitro gelation kinetics. The mechanical properties, drug delivery tests, and rheology tests suggest that PVA-MA derivatives have the potential to become a useful material for vitreous substitution. PMID- 15530062 TI - Adhesion behavior of peritoneal cells on the surface of self-assembled triblock copolymer hydrogels. AB - Adhesion behavior of cells to the surface of physical hydrogel membranes prepared by water-induced self-organization of precisely synthesized ABA-triblock copolymers comprised of poly(beta-benzyl L-aspartate) (PBLA) as A segment and poly(ethylene oxide) (PEO, molecular weight = 20 000) as the B segment were investigated. The cast film from the methylenechloride solution of these copolymers swelled in water very rapidly forming hydrogels (100-400% water content of total weight). The content of PBLA affected the strength, the hydrophobicity, and the amount of water involved in the hydrogel surface. During the early stage of cultivation with murine peritoneal cells, cell adhesion on the hydrogels of PEO and PBLA with 18 (20K18) and 25 (20K25) monomeric units was not observed, while adhesion on the hydrogels of PEO and PBLA with 32 (20K32) and 55 (20K55) monomeric units was successful, suggesting more than 12 mol % in PBLA content is necessary for adhesion of these cells. Although cell spreading on the hydrogels of 20K18, 20K25, and 20K32 was not sufficient, the hydrogel of 20K55 allowed cell adhesion and spreading to be bipolar with leading edge whose raffling is active with pseudopodium and lamellipodium as well as PBLA homopolymer, suggesting active motility of these cells. Remarkably, prolonged incubation restored adhesiveness onto the films at 20K18 in contrast to adhesion with 20K25 despite low hydrophobicity. It is conceivable that adaptation of proteins and chemical changes to the surface during the culture period may participate in these phenomena. Mechanical properties and interaction between cell and these copolymer hydrogels could be controlled by composition of block segments, and optimization for implants could also be attainable. PMID- 15530063 TI - Extent of charge screening in aqueous polysaccharide solutions. AB - The absorption optical system of a Beckman XL-I ultracentrifuge has been used to monitor the Donnan distribution of ions in polysaccharide solutions dialyzed against sodium phosphate buffer (pH 6.8, I 0.08) supplemented with 0.2 mM chromate as an indicator ion. For dextran sulfate, heparin, and polygalacturonate, the effective net charges are shown to be only one-third of those deduced from the chemical structures--a reflection of charge screening (counterion condensation) in aqueous polyelectrolyte solutions. Whereas the extent of charge screening for the first two polysaccharides agrees well with theoretical prediction, the disparity in the corresponding comparison for polygalacturonate reflects partial esterification of carboxyl groups, whereupon the experimental parameter refers to the effective charge per hexose residue rather than the effective fractional charge of each carboxyl group. PMID- 15530065 TI - Microbial synthesis of poly(3-hydroxyalkanoates) by Pseudomonas aeruginosa from fatty acids: identification of higher monomer units and structural characterization. AB - Pseudomonas aeruginosa ATCC 27853 accumulated poly(3-hydroxyalkanoates) (PHAs) after growth on saturated fatty acids with an odd number of carbon atoms. No nutrient limitation was required to induce PHA synthesis, although better yields were obtained when the medium was magnesium deprived. A comparative study was carried out between PHAs obtained from C-odd and those from C-even carbon sources. Repeating units identification was performed by gas chromatography (GC) and capillary liquid chromatography-electrospray mass spectrometry (LC-ESI MS) of methanolyzed samples. When C-odd n-alkanoic acids from nonanoic to pentadecanoic were used the lowest hydroxyalkanoate unit found was 3-hydroxyvalerate and the highest 3-hydroxypentadecanoate, whereas when C-even acids from octanoic to eicosanoic were used these were 3-hydroxycaproate and 3-hydroxyeicosanoate, respectively. Weight average molecular weights were in the range 187 000-596 000. DSC traces showed Tm and DeltaHm which varied from 43 to 58 degrees C and from 5.9 to 24.8 J/g, with the PHAs generated from C-odd carbon sources having lower values. ESI MS of partially pyrolyzed samples allowed the identification of oligomers up to heptamers, and statistical analysis of the ions intensity in the mass spectra showed that these PHAs are random copolyesters. PMID- 15530064 TI - Monodisperse chitosan nanoparticles for mucosal drug delivery. AB - Chitosan nanoparticles (CS NPs) of a controlled size (below 100 nm) and narrow size distribution were obtained through the process of ionic gelation between CS and sodium tripolyphosphate (TPP). A high degree of CS deacetylation and narrow polymer molecular weight distribution were demonstrated to be critical for the controlling particle size distribution. Properties of the CS NPs were examined at different temperatures, values of pH, and ratios of CS to TPP. The model protein, bovine serum albumin, was encapsulated into the NPs, and the in vitro release profiles were examined in physiologically relevant media at 37 degrees C. PMID- 15530066 TI - Synthesis and characterization of a photo-cross-linked biodegradable elastomer. AB - The synthesis and characterization of a photocurable biodegradable elastomer as a potential biomaterial for the delivery of thermosensitive drugs are described. The elastomer was prepared from UV initiated cross-linking of an acrylated star poly(epsilon-caprolactone-co-D,L-lactide) prepolymer. The influence of the molecular weight of the acrylated prepolymer on the final elastomer mechanical and thermal properties was determined. The glass-transition temperature of the elastomers was independent of the prepolymer molecular weight and was from -6 to 8 degrees C. The Young's modulus and stress at break of the elastomers was proportional to the inverse of the prepolymer molecular weight, while the strain at break increased in a linear fashion with the prepolymer molecular weight. Over a degradation period of 12 weeks in phosphate buffered saline, the elastomers exhibited little mass loss, appreciable mechanical strength loss, and little dimensional or strain at break change. PMID- 15530067 TI - PAMAM-PEG-PAMAM: novel triblock copolymer as a biocompatible and efficient gene delivery carrier. AB - A novel triblock copolymer, PAMAM-block-PEG-block-PAMAM was synthesized and applied as a gene carrier. PAMAM dendrimer is proven to be an efficient gene carrier itself, but it is associated with certain problems such as low water solubility and considerable cytotoxicity. Therefore, we introduced PEG to engineer a nontoxic and highly transfection efficient polymeric gene carrier because PEG is known to convey water-solubility and biocompatibility to the conjugated copolymer. This copolymer could achieve self-assembly with plasmid DNA, forming compact nanosized particles with a narrow size distribution. Fulfilling our expectations, the copolymer was found to form highly water-soluble polyplexes with plasmid DNA, showed little cytotoxicity despite its poor degradability, and finally achieved high transfection efficiency comparable to PEI in 293 cells. Consequently, these data show that an approach involving the introduction of PEG to create a tree-like cationic copolymer possesses a great potential for use in gene delivery systems. PMID- 15530068 TI - Antibacterial properties of nitric oxide-releasing sol-gel microarrays. PMID- 15530070 TI - Reversible electrochemical conversion between Rh(II) and Rh(III) states in Rh porphyrin adsorbed on carbon black. AB - We have clearly demonstrated reversible cyclic voltammograms for the redox reaction between the Rh(II) and Rh(III) states in rhodium octaethylporphyrin [Rh(OEP)] adsorbed on carbon black in an acidic aqueous solution. The emergence of the reversible wave can be ascribed to the suppression of the undesirable reactions between two molecules of [Rh(II)(OEP)] because of its strong interaction with the carbon black. The generated [Rh(II)(OEP)] exhibits a potent catalytic O(2) reduction activity. PMID- 15530069 TI - Photoinitiated DNA Binding by cis-[Ru(bpy)2(NH3)2]2+. AB - The ligand-loss photochemistry of cis-[Ru(bpy)(2)(NH(3))(2)](2+) (bpy = 2,2' bipyridine) was investigated in water and in the presence of added ligands such as bipyridine and chloride. Irradiation of the complex results in the covalent binding to 9-methyl- and 9-ethylguanine, as well as to single-stranded and double stranded DNA. This photoinduced DNA binding is not observed for the control complex [Ru(bpy)(2)(en)](2+) (en = ethylenediamine) under similar irradiation conditions. The results presented here show that octahedral Ru(II) complexes with photolabile ligands may prove useful as photoactivated cisplatin analogs. PMID- 15530071 TI - Pentadecadentate chelating ligands as building blocks for a [Fe6] cage with 12 exo-coordinated sodium cations. AB - Complexation of the highly branched, pentadecadentate chelating ligand cis,cis 1,3,5-cyclohexanetriamine-N,N,N',N',N",N"-hexaacetic acid (H(6)L) with iron(III) and sodium cations in the presence of carbonate anions leads to the formation of an [Fe6L2] cluster comprising an [Fe6] cage linked by 12 exo-coordinated sodium cations to form an extended 3D array. PMID- 15530072 TI - Structure and magnetic properties of a giant Cu44II aggregate which packs with a zeotypic superstructure. AB - Reaction of Cu(II) and the aminopolycarboxylate nitrilotripropionic acid (H(3)ntp) in water leads to the formation of [Cu(44)(mu(8)-Br)(2)(mu(3) OH)(36)(mu-OH)(4)(ntp)(12)Br(8)(OH(2))(28)]Br(2).81H(2)O. The Cu(44) aggregates have a central inorganic core corresponding to [Cu(24)(mu(8)-Br)(2)(mu(3) OH)(24)(mu-OH)(8)](14+) anchored on two bromide anions, and this is encased in a shell of Cu(II)/ligand units. The aggregates pack into a distorted tetragonal array with a very open structure containing large amounts of water of crystallization. The magnetic properties have been studied and, while complicated by the presence of low-lying excited states, indicate that the individual clusters have nonzero spin ground states. PMID- 15530073 TI - Synthesis, characterization, and reactivity of new copper(II) complexes of 2 methylthio-N-(2-pyridylmethyl)acetamide. AB - Copper(II) complexes were prepared with the new N(2)S(thioether) ligand 2 methylthio-N-(2-pyridylmethyl)acetamide (2-HL(N2S)). [Cu(2-L(N2S))Cl(MeOH)], which formed in the presence of excess triethylamine, is a distorted square pyramidal complex containing the ligand with the amide nitrogen deprotonated. The structurally analogous complex, [Cu(2-HL(N2S))Cl(2)], which formed in the absence of triethylamine, contains 2-HL(N2S) in the tautomeric imidic acid form. Neutral copper(II) N(2)S(thioether)S(thiolate) species were generated by addition of alkyl or aromatic thiolates to [Cu(2-L(N2S))Cl(MeOH)] and an unusual decomposition pathway was discovered. PMID- 15530074 TI - Ditopic double pincer palladacycle catalyst for C-C coupling. AB - A ditopic palladacycle with SCS pincer coordination, L(PdCl)(2) (1), was isolated and structurally characterized and represents the first example of a transition metal complex with a polythioamide-based macrocycle. Preliminary studies of 1 in the catalytic coupling of 4-iodotoluene and styrene indicated it to be robust in the presence of oxygen and high temperatures, with high turnover numbers in relatively short times. PMID- 15530075 TI - Gas-phase and solution-phase polymerization of epoxides by Cr(salen) complexes: evidence for a dinuclear cationic mechanism. AB - The gas-phase reactions of a series of mass-selected mononuclear and dinuclear Cr(salen) complexes with propylene oxide suggest that the enhanced reactivity of the dinuclear complexes in gas-phase and in solution may derive from a dicationic mechanism in which the alkoxide chain is mu(2)-coordinated to two Lewis acidic metal centers. The double coordination is proposed to suppress backbiting, and hence chain-transfer in the gas-phase homopolymerization of epoxides. PMID- 15530076 TI - Modeling the peroxide/superoxide continuum in 1:1 side-on adducts of O2 with Cu. AB - The character of singlet (C(3)N(2)H(5))CuO(2) ranges smoothly between copper(III) peroxide and copper(II) superoxide with variation of the electronic character of the supporting beta-diketiminate ligand. Over the range of the variation, multireference second-order perturbation theory predicts the (1)A(1) singlet state always to be lower in energy than the lowest triplet state ((3)B(1)). The multideterminantal character of the biradical-like superoxide mesomer causes density functional theory sometimes to fail badly in predicting the relative energies of these same states, although its predictions of other properties, such as geometry, are of good quality. PMID- 15530077 TI - The renaissance of aluminum chemistry. AB - The preparation and reactions of stable monomeric and tetrameric organoaluminum(I) compounds are described. Especially, their conversion to the more stable oxidation state (III) is discussed. The synthesis of unusual and unique molecules such as LAl(OH)(2), LAl(SH)(2), LAl(SeH)(2), and LAl(NH(2))(2) is reported. A new synthetic approach to organometallic hydroxides and amides was discovered, when N-heterocyclic carbenes are used as hydrogen chloride acceptors. The synthesis of LGeOH demonstrates that this method can also be applied for other elements in the periodic system. The successful preparation of carbaalanes and aluminum nitride clusters resulted in the generation of several model compounds for the fixation of organometal substituents on carbide and nitride surfaces. It is shown that AlH(3).NMe(3) is a powerful hydroalumination reagent for acetylenes, nitriles, and isonitriles. PMID- 15530078 TI - New 1:3 type nickel-bis(dithiolene) salt (FcCHCHPymCH3)[Ni(dmit)2]3 (dmit: 2 thioxo-1,3-dithiole-4,5-dithiolate): its electrocrystallization, crystal structure, electrical properties, and electronic band structure analysis. AB - Black single crystals of Ni(dmit)(2) complex (dmit: 2-thioxo-1,3-dithiole-4,5 dithiolate) with trans-4-[2-(1-ferrocenyl)vinyl]-1-methylpyridinium chromophore as a countercation, (FcCHCHPymCH(3))[Ni(dmit)(2)](3), were prepared by the electrocrystallization technique. In the triclinic structure of the complex (P, a = 11.430(5) A, b = 13.349(2) A, c = 19.355(6) A, alpha = 75.15(2) degrees , beta = 79.19(3) degrees , gamma = 82.12(2) degrees , Z = 2), Ni(dmit)(2) anion layers are separated by the cations with a relatively rare 1:3 cation-to-anion ratio. Detailed crystal and electronic structure analysis revealed that the anions are stacked in the layers to form alternating dimers and monomers rather than trimers. The measured electrical conductivity indicates a semiconducting property of the compound with an estimated energy gap of 0.06 eV. The calculated LUMO bands are very narrow, and the semiconducting behavior is more likely due to the electron localization mainly on the dimers, consistent with the observed longer Ni-S bond distances in the dimers. PMID- 15530079 TI - Synthesis, structure, and cooperative proton-electron transfer reaction of Bis(5,6-diethylpyrazinedithiolato)metal complexes (M = Ni, Pd, Pt). AB - New proton and electron donors, M(II)(HL)(2) (M = Ni, Pd, Pt; L = 5,6 diethylpyradzinedithiolate), as well as a proton and electron acceptor, Pt(IV)(L)(2), were prepared and characterized. The pH-dependent cyclic voltammetry of the M(II)(HL)(2) complexes revealed a favorable Gibbs free energy (K(com) > 1) for the proton and electron transfer reactions from M(II)(HL)(2) to M(IV)(L)(2); i.e., the equilibrium for the following reaction lies to the right: M(II)(HL)(2) + M(IV)(L)(2) <==>2M(III)(HL)(L). PMID- 15530080 TI - DOTP-manganese and -nickel complexes: from a tetrahedral network with 12-membered rings to an ionic phosphonate. AB - DOTP (1,4,7,10-tetrakis(methylenephosphonic acid)-1,4,7,10-tetraazacyclododecane) was reacted hydrothermally with MnCl(2).2H(2)O and Ni(NO(3))(2).6H(2)O resulting in two structurally different compounds. Mn[C(3)NH(7)(PO(3)H(0.5))](4) crystallizes in the tetragonal space group P4/ncc, with a = 12.349(2) A, b = 12.349(2) A, c = 14.066(4) A, V = 2144.9(8) A(3), and Z = 4. Manganese atoms are tetrahedrally bonded by four phosphonate oxygen atoms from four equivalent ligands. All 12-membered macrocycles are connected in a "zigzag" manner by sharing manganese atoms and forming 22-membered cavities between each pair of two adjacent macrocycles. Ni[C(3)NH(6)(PO(3)H)](4)[Ni(H(2)O)(6)] crystallizes as an ion pair complex. Ni(1) is octahedrally coordinated to two pendent phosphonate oxygen atoms and four nitrogen atoms from the macrocyclic backbone. Ni(2) is surrounded by six coordinatedly bonded water molecules to form a hexaqua cation. The manganese complex shows ion exchange capability for Cs(+). PMID- 15530081 TI - A family of Mn16 single-molecule magnets from a reductive aggregation route. AB - The synthesis and magnetic properties of three isostructural hexadecametallic manganese clusters [Mn(16)O(16)(OMe)(6)(O(2)CCH(2)Ph)(16)(MeOH)(6)] (1), [Mn(16)O(16)(OMe)(6)(O(2)CCH(2)Cl)(16)(MeOH)(6)] (2), and [Mn(16)O(16)(OMe)(6)(O(2)CCH(2)Br)(16)(MeOH)(6)] (3) are reported. The complexes were prepared by a reductive aggregation reaction involving phenylacetic acid, chloroacetic acid or bromoacetic acid, and NBu(n)()(4)MnO(4) in MeOH. Complex 1 crystallizes in the monoclinic space group C2/c and consists of 6 Mn(IV) and 10 Mn(III) ions held together by 14 mu(3)-O(2)(-), 2 mu-O(2)(-), 4 mu-MeO(-), and 2 mu-O(2)CCH(2)Ph(-) groups. The remaining 14 mu-O(2)CCH(2)Ph(-) ligands, 2 mu-MeO( ) groups, and 6 terminal MeOH molecules constitute the peripheral ligation in the complex. Variable-temperature, solid-state dc magnetic susceptibility measurements on 1-3 in the temperature range 5.0-300 K reveal that all three complexes are dominated by intramolecular antiferromagnetic exchange interactions. Low-lying excited states preclude an exact determination of the spin ground state for 1-3 by magnetization measurements. Alternating current susceptibility measurements at zero dc field in the temperature range 1.8-10 K and a 3.5 G ac field oscillating at frequencies in the 5-1488 Hz range display, at temperatures below 3 K, a nonzero, frequency-dependent chi(M)"signal for complexes 1-3, with the peak maxima lying at temperatures less than 1.8 K. Single crystal magnetization versus dc field scans down to 0.04 K for complex 1 show hysteresis behavior at <1 K, establishing 1 as a new member of the SMM family. No clear steps characteristic of quantum tunneling of magnetization (QTM) were observed in the hysteresis loops. PMID- 15530082 TI - Nonoxovanadium(IV) and oxovanadium(V) complexes with mixed O, X, O-donor ligands (X = S, Se, P, or PO). AB - Ligating properties of four potentially tridentate bisphenol ligands containing [O, X, O] donor atoms (X = S 1, Se 2, P 3, or P=O 4) toward the vanadium ions in +IV or +V oxidation states have been studied. Each ligand with different heterodonor atoms yields as expected nonoxovanadium(IV) complexes, V(IV)L(2), whose structures have been determined by X-ray diffraction methods as having six coordinate V(IV), VO(4)X(2), core. Compounds 1-4 have also been studied with electrochemical methods, variable-temperature (2-295 K) magnetic susceptibility measurements, X-band electron paramagnetic resonance (EPR) (2-60 K) spectroscopy, and magnetic circular dichroism (MCD) (5 K) measurements. Electrochemical results suggest metal-centered oxidations to V(V) (i.e., no formation of phenoxyl radicals from the coordinated phenolates). A combination of density functional theory calculations and experimental EPR investigations indicates a dramatic effect of the heteroatoms on the electronic structure of 1-4 with consequent reordering of the energy levels; 1 and 3 possess a trigonal ground state (d(z)()(2))(1), but 4 with the phosphoryl oxygen as the heterodonor atom in contrast exhibits a tetragonal ground state, (d(xy)())(1). On the basis of the intense electronic transitions in absorption spectra, all electronic transitions observed for 4 have been assigned to ligand-to-metal charge-transfer transitions, which have been confirmed by preliminary resonance Raman measurements and C/D ratios obtained from low-temperature MCD spectroscopy. Moreover, diamagnetic complexes 5 and 6 containing mononuclear and dinuclear oxovanadium(V) units have also been synthesized and structurally and spectroscopically ((51)V NMR) characterized. PMID- 15530083 TI - Size and surface effects of prussian blue nanoparticles protected by organic polymers. AB - Prussian blue (PB) nanoparticles protected by organic polymers such as poly(vinylpyrrolidone) (PVP) and poly(diallyldimethylammonium chloride) (PDDA) were prepared. Different experimental conditions (concentrations of Fe ions and feed ratios of Fe to the polymers) have been investigated to control the size of the PB nanoparticles. For example, the averaged dimensions of the PB nanoparticles were tuned from 12 to 27 nm by use of PVP in the different conditions. Addition of PDDA produced the PB nanoparticles with very small dimensions (5-8 nm) by an effective electrostatic interaction. We found that the surface environments of the PB nanoparticles affect the inherent properties of PB. The shifts of charge transfer (CT) absorptions from Fe(2+) to Fe(3+) in the PB nanoparticles resulted from the surface-protecting polymers. Especially, the PB nanoparticles with the PVP protection show high solubility in a variety of organic solvents and a solvent-dependent CT absorption. Measurement of the magnetic properties of the PB nanoparticles showed unprecedented size-dependency, surface effect, and superparamagnetic properties. PMID- 15530084 TI - Terphenyl ligand stabilized lead(II) derivatives: steric effects and lead-lead bonding in diplumbenes. AB - The reaction of PbBr(2) with the lithium reagents LiC(6)H(3)-2,6-(C(6)H(3)-2,6 Pr(i)(2))(2) (LiArPr(i)(2)) and Et(2)O.LiC(6)H(3)-2,6-(2,6-Pr(i)-4 Bu(t)C(6)H(2))(2) (Et(2)O.LiArPr(i)(2)Bu(t)) furnished the bromide bridged organolead(II) halides [Pb(mu-Br)ArPr(i)(2)](2) (1) and[Pb(mu Br)ArPr(i)(2)Bu(t)](2) (2) as orange crystals. Treatment of 1 with a stoichiometric amount of methylmagnesium bromide resulted in the "diplumbene" Pr(i)(2)Ar(Me)PbPb(Me)ArPr(i)(2) (3). The addition of 1 equiv of 4-tert butylphenylmagnesium bromide to 1 afforded the feebly associated, Pb-Pb bonded species [Pb(C(6)H(4)-4-Bu(t))ArPr(i)(2)](2) (4), whereas the corresponding reaction of tert-butylmagnesium chloride and 1 afforded the monomer Pb(Bu(t))ArPr(i)(2) (5). The reaction of the more crowded aryl lead(II) bromide [Pb(mu-Br)ArPr(i)(3)](2) (Ar = C(6)H(3)-2,6(C(6)H(2)-2,4,6-Pr(i)(3))(2)) with 4 isopropyl-benzylmagnesium bromide or LiSi(SiMe(3))(3) yielded the monomers 6, [Pb(CH(2)C(6)H(4)-4-Pr(i))ArPr(i)(3)], or 7, [Pb(Si(SiMe(3))(3))ArPr(i)(3)]. All compounds were characterized with use of X-ray crystallography, (1)H, (13)C, and (207)Pb NMR (3-7), and UV-vis spectroscopy. The dimeric Pb-Pb bonded (Pb-Pb = 3.1601(6) A) structure of 3 may be contrasted with the previously reported monomeric structure of Pb(Me)ArPr(i)(3), which differs from 3 only in that it has para Pr(i) substituents on the flanking aryl rings. The presence of these groups is sufficient to prevent the weak Pb-Pb bonding seen in 3. The dimer 4 displays a Pb-Pb distance of 3.947(1) A, which indicates a very weak lead-lead interaction, and it is possible that this close approach could be caused by packing effects. The monomeric structures of 6 and 7 are attributable to steric effects and, in particular, to the large size of ArPr(i)(3). PMID- 15530085 TI - Multi-iron silicotungstates: synthesis, characterization, and stability studies of polyoxometalate dimers. AB - The reaction of Fe(III) with Na(+) and K(+) salts of the trivacant [alpha SiW(9)O(34)](10)(-) ligand have been investigated at pH 6 and pH 1. A new dimer, [(alpha-SiFe(3)W(9)(OH)(3)O(34))(2)(OH)(3)](11-) (1), is synthesized by reacting Na(7)H(3)[alpha-SiW(9)O(34)] or K(10)[alpha-SiW(9)O(34)] with exactly 3 equiv of Fe(III) in a 0.5 M sodium acetate solution (pH 6). The structure of 1, determined by single-crystal X-ray diffraction (a = 22.454(2) A, b = 12.387(2) A, c = 37.421(2), beta = 100.107(8) degrees , monoclinic, C2/c, Z = 4, R(1) = 5.11% based on 12739 independent reflections), consists of two [alpha SiFe(3)W(9)(OH)(3)O(34)](4-) units linked by three Fe-mu-OH-Fe bonds. Reaction of K(10)[alpha-SiW(9)O(34)] with 3 equiv of Fe(III) in water (pH 1) yields [(alpha Si(FeOH(2))(2)FeW(9)(OH)(3)O(34))(2)](8)(-2). The structure of 2 was also determined by single-crystal X-ray diffraction (a = 36.903(2) A, b = 13.9868(9) A, c = 21.7839(13) A, beta = 122.709(1) degrees , monoclinic, C2/c, Z = 4, R(1) = 4.57% based on 11787 independent reflections). It consists of two [alpha Si(FeOH(2))(2)FeW(9)(OH)(3)O(34)](4-) Keggin units linked by a single edge. The terminal ligand on Fe1 in each trisubstituted Keggin unit becomes a mu(2) oxo ligand bridging to a [WO(6)](2-) moiety. The UV-vis spectra of both complexes show the characteristic oxygen-to-metal-charge-transfer bands of polyoxometalates as well as an Fe(III)-centered band at 436 nm (epsilon = 146 M(-1) cm(-1)) and 456 nm (epsilon = 104 M(-1) cm(-1)) for complexes 1 and 2, respectively. Differential scanning calorimetry data show that complex 1 decomposes between 575 and 600 degrees C whereas no decomposition is observed for complex 2 up to temperatures of 600 degrees C. PMID- 15530086 TI - Mononuclear and binuclear wirelike ruthenium(II) complexes with oligo-diethynyl thiophene bridged back-to-back terpyridine ligands: synthesis and electrochemical and photophysical properties. AB - The syntheses, structural features, electrochemical behavior, absorption spectra, and photophysical properties of five mononuclear complexes [(terpy)Ru(terpy DEDBT(n)-terpy)](2+), RuT(n), and five binuclear complexes [(terpy)Ru(terpy DEDBT(n)-terpy)Ru(terpy)](4+), RuT(n)Ru, are reported, where n varies from 1 to 5 so that the metal-metal distance is estimated to be 42 A for the largest binuclear complex, RuT(5)Ru (terpy is 2,2':6',2"-terpyridine and DEDBT is 2,5 diethynyl-3,4-dibutylthiophene). The metal-centered oxidation potentials for the mononuclear and binuclear species are slightly more positive than for the reference [Ru(terpy)(2)](2+) complex, owing to the withdrawing nature of the back to-back terpyridine ligands incorporating the repeat diethynyl-thiophene units. Comparison of the reduction potentials for the mononuclear and binuclear complexes reveals that the reduction steps are localized either at the terpy fragments of the T(n) ligands or at the terpy peripheral ligands. The spectroscopic results (absorption spectra at room temperature, luminescence spectra and lifetimes at room temperature and at 77 K) in acetonitrile solvent are consistent with the establishment of electronic delocalization within the oligomeric diethynyl-thiophene fragments (DEDBT(n)) of the T(n) ligands; however, the results also indicate that the terpy units of these ligands and the DEDBT(n)fragments are not strongly coupled. Both at room temperature and at 77 K, the (3)metal-to-ligand charge-transfer luminescence of RuT(n) and RuT(n)Ru complexes is strongly depressed in the larger species with respect to what happens for n < or = 2 (where the luminescence quantum yield is phi approximately 10(-4)); this is discussed in terms of the possible intervention of triplet levels localized at the oligothiophene DEDBT(n)(fragments. PMID- 15530087 TI - Influence of anionic functions on the coordination and photophysical properties of lanthanide(III) complexes with tridentate bipyridines. AB - A series of four ligands based on a 5'-methyl-2,2'-bipyridyl framework substituted in the 6 position by a carboxylic acid, a phosphonic acid, a monoethyl ester phosphonic acid, or a diethyl ester phosphonic acid are described. The pK(a) values of all ligands and their assignments are determined by a combination of UV-vis absorption spectroscopy and (1)H and (31)P NMR spectroscopy. The ability of the tridentate ligands to form complexes with trivalent lanthanide cations (Ln = La, Nd, Eu, and Lu) in buffered water solutions (Tris-HCl, pH = 7.4) is studied by UV-vis absorption spectroscopy and (1)H NMR. While the two ester ligands display a weak coordination ability toward lanthanide cations, the acid ligands form stable complexes with 1:1, 1:2, and 1:3 Ln/L ratios. A weak selectivity is observed for the middle of the lanthanide series, and the complexes of the phosphonic acid derivative are up to 2 orders of magnitude more stable than those of the carboxylic acid ligand. Photophysical properties of the free phosphonic and carboxylic acid ligands and of their complexes with La, Eu, Gd, Tb, and Lu are investigated in buffered aqueous solutions both at room temperature and 77 K. An efficient ligand-to-metal energy transfer is observed for both the Eu and Tb complexes. Despite a relatively large energy gap between the ligand-centered (3)pipi* and the Eu((5)D(0)) or Tb((5)D(4)) emitting states, the metal-centered luminescence is well sensitized with quantum yields reaching up to 45.5 and 42.2% for the Tb 1:3 complexes with carboxylic and phosphonic acid ligands, respectively. PMID- 15530088 TI - Ultrafast photochemical dissociation of an equatorial CO ligand from trans(X,X) [Ru(X)2(CO)2(bpy)] (X = Cl, Br, I): a picosecond time-resolved infrared spectroscopic and DFT computational study. AB - Ultrafast photochemistry of the complexes trans(X,X)-[Ru(X)(2)(CO)(2)(bpy)] (X = Cl, Br, I) was studied in order to understand excited-state reactivity of equatorial CO ligands, coordinated trans to the 2,2'-bipyridine ligand (bpy). TD DFT calculations have identified the lowest electronic transitions and singlet excited states as mixed X -->bpy/Ru --> bpy ligand to ligand/metal to ligand charge transfer (LLCT/MLCT). Picosecond time-resolved IR spectroscopy in the region of nu(CO) vibrations has revealed that, for X = Cl and Br, subpicosecond CO dissociation is accompanied by bending of the X-Ru-X moiety, producing a pentacoordinated intermediate trans(X,X)-[Ru(X)(2)(CO)(bpy)]. Final movement of an axial halide ligand to the vacant equatorial position and solvent (CH(3)CN) coordination follows with a time constant of 13-15 ps, forming the photoproduct cis(X,X)-[Ru(X)(2)(CO)(CH(3)CN)(bpy)]. For X = I, the optically populated (1)LLCT/MLCT excited state undergoes a simultaneous subpicosecond CO dissociation and relaxation to a triplet IRuI-localized excited state which involves population of an orbital that is sigma-antibonding with respect to the axial I-Ru I bonds. Vibrationally relaxed photoproduct cis(I,I)-[Ru(I)(2)(CO)(CH(3)CN)(bpy)] is formed with a time constant of ca. 55 ps. The triplet excited state is unreactive, decaying to the ground state with a 155 ps lifetime. The experimentally observed photochemical intermediates and excited states were assigned by comparing calculated (DFT) and experimental IR spectra. The different behavior of the chloro and bromo complexes from that of the iodo complex is caused by different characters of the lowest triplet excited states. PMID- 15530089 TI - Modulation of molybdenum-centered redox potentials and electron-transfer rates by sulfur versus oxygen ligation. AB - Temperature-dependent measurements of potential, E degrees', and electron transfer rate constant, k(s,h), are reported for electrochemical reduction (in 0.3 M TBAPF(6)/CH(3)CN) of a series of oxomolybdenum(V) complexes, [(Tp)MoO(X,Y)], where Tp = hydrotris(3,5-dimethyl-1-pyrazolyl)borate and X,Y is a series of bidentate 1,2-disubstituted aliphatic or aromatic ligands in which oxygen donors are replaced sequentially by sulfur. E degrees' values shift in the positive direction, and k(s,h) values increase as O is replaced by S and as the framework of the ligand is changed from aliphatic to aromatic. The electrochemical enthalpy of activation, measured under conditions of zero driving force as DeltaH= -R partial differential[ln(k(s,h))]/ partial differential(1/T) and corrected for an outer-shell component by the mean spherical approximation, is approximately 10 kJ mol(-1) larger for complexes with O versus S donors and with an aliphatic versus aromatic ligand framework. Thus, the rate of Mo(V/IV) electron transfer is modulated primarily by differences in inner-shell reorganization. Following a recent description of electronic structure contributions to electron-transfer reactivity (Kennepohl, P.; Solomon, E. I. Inorg. Chem. 2003, 42, 679 ff), it is concluded that more effective charge distribution over the entire molecular structure, as mediated by electronic relaxation in S versus O and aromatic versus aliphatic systems, is responsible for the influence of ligand structure on the kinetics and thermodynamics of Mo centered electron transfer. There is no evidence, based on experimentally measured pre-exponential factors, that sulfur donors or an aromatic ligand framework are more effective than their structural counterparts in facilitating electronic coupling between the electrode and the Mo d(xy) redox orbital. PMID- 15530090 TI - Silver(I) complexes of a sterically demanding fluorinated triazapentadienyl ligand [N((C3F7)C(Dipp)N)2]- (Dipp = 2,6-diisopropylphenyl). AB - Sterically demanding triazapentadiene [N((C3F7)C(Dipp)N)2]H affords the isolation of thermally stable, two- and three-coordinate silver complexes. The free ligand [N((C3F7)C(Dipp)N)2]H has a W-shaped ligand backbone in the solid state.[N((C3F7)C(Dipp)N)2]H reacts with silver(I) oxide in acetonitrile leading to CH(3)CNAg [N((C3F7)C(Dipp)N)2]HIt features a two-coordinate silver center and a kappa(1)-coordinated triazapentadienyl ligand. This silver acetonitrile complex serves as an excellent precursor to obtain thermally stable, silver isocyanide t BuNCAg [N((C3F7)C(Dipp)N)2]Hand silver phosphine [[N((C3F7)C(Dipp)N)2]HAgPPh(3) adducts. IR spectroscopic data for the silver(I) isocyanide t-BuNCAg [N((C3F7)C(Dipp)N)2]Hshows nu(CN) at 2219 cm(-)(1). The silver ion coordinates to the triazapentadienyl ligand via the central nitrogen atom. The silver PPh(3) adduct,[N((C3F7)C(Dipp)N)2]HAgPPh(3), was synthesized by treating CH3CNAg [N((C3F7)C(Dipp)N)2]Hwith PPh(3). It displays relatively large Ag-P coupling in the (31)P NMR spectrum. The triazapentadienyl ligand in[N((C3F7)C(Dipp)N)2]HAgPPh(3) acts as a chelating kappa(2)-donor. The Ag-P bond is relatively short (2.3487(10) A). PMID- 15530092 TI - Kinetics and mechanisms of bromine chloride reactions with bromite and chlorite ions. AB - Chloride ion catalyzes the reactions of HOBr with bromite and chlorite ions in phosphate buffer (p[H(+)] 5 to 7). Bromine chloride is generated in situ in small equilibrium concentrations by the addition of excess Cl(-) to HOBr. In the BrCl/ClO(2)(-) reaction, where ClO(2)(-) is in excess, a first-order rate of formation of ClO(2) is observed that depends on the HOBr concentration. The rate dependencies on ClO(2)(-), Cl(-), H(+), and buffer concentrations are determined. In the BrCl/BrO(2)(-) reaction where BrCl is in pre-equilibrium with the excess species, HOBr, the loss of absorbance due to BrO(2)(-) is followed. The dependencies on Cl(-), HOBr, H(+), and HPO(4)(2)(-) concentrations are determined for the BrCl/BrO(2)(-) reaction. In the proposed mechanisms, the BrCl/ClO(2)(-) and BrCl/BrO(2)(-) reactions proceed by Br(+) transfer to form steady-state levels of BrOClO and BrOBrO, respectively. The rate constant for the BrCl/ClO(2)( ) reaction [k(Cl)(2)]is 5.2 x 10(6) M(-1) s(-1) and for the BrCl/BrO(2)(-) reaction [k(Br)(2)]is 1.9 x 10(5) M(-1) s(-1). In the BrCl/ClO(2)(-) case, BrOClO reacts with ClO(2)(-) to form two ClO(2) radicals and Br(-). However, the hydrolysis of BrOBrO in the BrCl/BrO(2)(-) reaction leads to the formation of BrO(3)(-) and Br(-). PMID- 15530091 TI - Phosphorus-nitrogen donor interaction leading to atrane formation in phosphate and oxyphosphorane compositions. Implications for phosphoryl transfer enzymes(1). AB - Reaction of aminotriphenols, tris(2-hydroxy-3,5-dimethylbenzyl)amine (E) and tris(2- hydroxy-3-tert-butyl-5-methylbenzyl)amine (F), with triphenylphosphite, tris(p-methoxyphenyl)phosphite, or phenyldiphenoxyphosphane in the presence of N chlorodiisopropylamine led to the isolation of tetraoxyphosphorane 1, pentaoxyphosphorane 3, phosphate-atrane 2, hexacoordinated pentaoxyphosphorane atrane 4, and the first hexacoordinated tetraoxyphosphorane-atrane 5. X-ray analysis of 1-3 and 5 were obtained. NMR data is reported and supports that the atrane 4 has the same hexacoordinated structure as 5. Phosphate 2 reveals weak phosphorus-nitrogen donor action whereas the hexacoordinated atranes 4 and 5 have pronounced P-N coordination. The results are used to support amino acid donor action occurring at active sites of phosphoryl transfer enzymes. Increased strength of donor action in the higher-coordinate model activated state compared to that in the substrate phosphate composition should serve as a factor in enhancing enzyme reaction rates. PMID- 15530093 TI - Molecular design of luminescence ion probes for various cations based on weak gold(I)...gold(I) interactions in dinuclear gold(I) complexes. AB - A series of luminescent dinuclear gold(I) complexes with different crown ether pendants, [Au(2)(PwedgeP)(S-B15C5)(2)] [S-B15C5 = 4'-mercaptobenzo-15-crown-5, P(wedge)P = bis(dicyclohexylphosphino)methane (dcpm) (1), bis(diphenylphosphino)methane (dppm) (2)] and [Au(2)(P(wedge)P)(S-B18C6)(2)] [S B18C6 = 4'-mercaptobenzo-18-crown-6, P(wedge)P = dcpm (3), dppm (4)], and their related crown-free complexes, [Au(2)(P(wedge)P)(SC(6)H(3)(OMe)(2)-3,4)(2)] [P(wedge)P = dcpm (5), dppm (6)], were synthesized. The low-energy emission of the mercaptocrown ether-containing gold(I) complexes are tentatively assigned as originated from states derived from a S --> Au ligand-to-metal charge transfer (LMCT) transition. The crown ether-containing gold(I) complexes showed specific binding abilities toward various metal cations according to the ring size of the crown pendants. Spectroscopic evidence was provided for the metal-ion-induced switching on of the gold...gold interactions upon the binding of particular metal ions in a sandwich binding mode. PMID- 15530094 TI - An investigation of Staudinger reactions involving cis-1,3,5-triazidocyclohexane and tri(alkylamino)phosphines. AB - The reaction of 1,3,5-cis-triazidocyclohexane with the electron-rich tris(dialkylamino)phosphines P(NMe(2))(3) (1) and N(CH(2)CH(2)NMe)(3)P (2b) in acetonitrile for 3 h furnished the corresponding tris-phosphazides 1,3,5-cis (R(3)PN(3))(3)C(6)H(9), 3a (R(3)P = 1) and 3b (R(3)P = 2b), in 90% and 92% yields, respectively. The same reaction with the relatively electron-poor tris(dialkylamino)phosphine MeC(CH(2)NMe)(3)P (4) for 2 days gave the tris iminophosphorane, 1,3,5-cis-(R(3)PN)(3)C(6)H(9), 5a (R(3)P = 4), in 60% yield. Compound 3b is a thermally stable solid that did not lose dinitrogen when refluxed in toluene for 24 h or when heated as a neat sample at 100 degrees C /0.5 Torr for 10 h. By contrast, tris-phosphazide 3a decomposed to the tris iminophosphorane 1,3,5-cis-(R(3)PN)(3)C(6)H(9), 5b (R(3)P = 1), in 3 h in quantitative yield upon heating to 100 degrees C in toluene. Factors influencing the formation of the phosphazides or the iminophosphoranes in these reactions are discussed. The reaction of 3b with 4 equiv of benzoic acid gave [N(CH(2)CH(2)NMe)(3)P=NH(2)]PhCO(2) ([6bH]PhCO(2)) in quantitative yield along with benzene (56% yield) and dinitrogen. The same reaction with 3a gave [(Me(2)N)(3)P=NH(2)]PhCO(2) ([7aH]PhCO(2)) (quantitative yield), benzene (15% yield), and dinitrogen(.) Treatment of [6bH]PhCO(2) with KO(t)Bu afforded N(CH(2)CH(2)NMe)(3)P=NH (6b) in 40% overall yield. Compound 6b upon treatment with PhCH(2)CH(2)Br produced [6bH]Br in 90% yield along with styrene. The new compounds were characterized by analytical and spectroscopic methods, and selected compounds (3b, 5a, and [6bH]Br) were structured by X-ray crystallography. A special feature of 3b is its capability to function as a starting material for 6b, which was not accessible by other synthetic routes. PMID- 15530096 TI - Unusual reduction of ammonium heptamolybdate to novel molybdenum(IV)-stabilized azo anion radical complexes. AB - In an unusual reaction of the polyacid, ammoniumheptamolybdate tetrahydrate ((NH(4))(6)[Mo(7)O(24)].4H(2)O), and the ligand, 2-[(arylamino)phenylazo]pyridine (general abbreviation HL), in the presence of PPh(3) afforded the brown oxo free molybdenum complexes of type [Mo(L)(2)] in high yields (ca. 80%). The reaction occurs smoothly in ethanol. It is slow on a steam bath (25 h) but is complete in about an hour in a microwave oven. X-ray structures of two representatives are reported. In these complexes the ligand acted as a tridentate ligand using its pyridyl(N), azo(N), and the deprotonated amine(N), respectively. The geometry is meridional, and the relative orientations within the coordinated pairs of nitrogens are cis, trans, and cis, respectively. Bond length data of the coordinated ligands are consistent with a Mo(IV)[L.](2) ([L.](2-) = azo dianion radical formed by one electron reduction of the deprotonated anionic ligand, [L]( )) description. For example, the N-N lengths (1.349(5)-1.357(2) A) in these complexes are appreciably longer than that (1.246(3) A) in the uncoordinated and protonated salt of a representative ligand, [H(2)L(d)]ClO(4). The N-N lengths, however, correspond well with metal complexes of the ligand containing azo ion radical. The complexes are diamagnetic and showed highly resolved (1)H NMR and (13)C NMR spectra. The two coordinated ligands in these are magnetically equivalent, and resonances for only one ligand were observed in their spectra. Diamagnetism in the present molybdenum complexes is attributed to strong antiferromagnetic coupling between Mo(IV)(4d(2)) and the two planar radical [L.](2-) ligands. The complexes display multiple redox responses. The ESR spectrum of electrogenerated [1a](-) showed a characteristic spectrum for Mo(III) with weak hyperfine lines due to the presence of molybdenum isotopes having nonzero nuclear spin. Visible range multiple charge transfer transitions in these complexes are ascribed to ligand-to-metal transitions. PMID- 15530095 TI - Alkyl- and aryl-substituted corroles. 5. Synthesis, physicochemical properties, and X-ray structural characterization of copper biscorroles and porphyrin-corrole dyads. AB - The synthesis and characterization of cofacial copper biscorroles and porphyrin corroles linked by a biphenylenyl or anthracenyl spacer are described. The investigated compounds are represented as (BCA)Cu(2) and (BCB)Cu(2) in the case of the biscorrole (BC) derivatives and (PCA)Cu(2) and (PCB)Cu(2) in the case of porphyrin (P)-corrole (C) dyads, where A and B represent the anthracenyl and biphenylenyl bridges, respectively. A related monomeric corrole (Me(4)Ph(5)Cor)Cu and monomeric porphyrin (Me(2)Et(6)PhP)Cu that comprise the two halves of the porphyrin-corrole dyads were also studied. Electron spin resonance (ESR), (1)H NMR, and magnetic measurements data demonstrate that the copper corrole macrocycle, when linked to another copper corrole or copper(II) porphyrin, can be considered to be a Cu(III) complex in equilibrium with a Cu(II) radical species, copper(III) corrole being the main oxidation state of the corrole species at all temperatures. The cofacial orientation of (BCB)Cu(2), (BCA)Cu(2), and (PCB)Cu(2) was confirmed by X-ray crystallography. Structural data: (BCB)Cu(2)(C(110)H(82)N(8)Cu(2).3CDCl(3)), triclinic, space group P, a = 10.2550(2) A, b = 16.3890(3) A, c = 29.7910(8) A, alpha = 74.792(1) degrees , beta = 81.681(1) degrees , gamma = 72.504(2) degrees , Z = 2; (BCA)Cu(2)(C(112)H(84)N(8)Cu(2).C(7)H(8).1.5H(2)O), monoclinic, space group P 2(1)/c, a = 16.0870(4) A, b = 35.109(2) A, c = 19.1390(8) A, beta = 95.183(3) degrees , Z = 4; (PCB)Cu(2)(C(89)H(71)N(8)Cu(2).CHCl(3)), monoclinic, space group P2(1)/n, a = 16.7071(3) A, b = 10.6719(2) A, c = 40.8555(8) A, beta = 100.870(1) degrees , Z = 4. The two cofacial biscorroles, (BCA)Cu(2) and (BCB)Cu(2), both show three electrooxidations under the same solution conditions. The reduction of (BCA)Cu(2) involves a reversible electron addition to each macrocycle at the same potential of E(1/2) = -0.20 V although (BCB)Cu(2) is reversibly reduced in two steps to give first [(BCB)Cu(2)](-) and then [(BCB)Cu(2)](2)(-), each of which was characterized by ESR spectroscopy as containing a Cu(II) center. These latter electrode reactions occur at E(1/2) = -0.36 and -0.51 V versus a saturated calomel reference electrode. The half-reduced and fully reduced (BCB)Cu(2) show similar Cu(II) ESR spectra, and no evidence of a triplet signal is observed. The two well-separated reductions of (BCB)Cu(2) to give [(BCB)Cu(2)](2)(-) can be attributed to a stronger pi-pi interaction between the two macrocycles of this dimer as compared to those of (BCA)Cu(2). The copper porphyrin-corrole dyads, (PCA)Cu(2) and (PCB)Cu(2), show five reversible oxidations and two reversible reductions, and these potentials are compared with corresponding values for electrochemical reactions of the porphyrin and corrole monomers under the same solution conditions. PMID- 15530097 TI - Binding of the oxo-rhenium(V) core to methionine and to N-terminal histidine dipeptides. AB - The ReOX(2)(met) compounds (X = Cl, Br) adopt a distorted octahedral structure in which a carboxylato oxygen lies trans to the Re=O bond, whereas the equatorial plane is occupied by two cis halides, an NH(2), and an SCH(3) group. Coordination of the SCH(3) unit creates an asymmetric center, leading to two diastereoisomers. X-ray diffraction studies reveal that the crystals of ReOBr(2)(d,l-met).1/2H(2)O and ReOBr(2)(d,l-met).1/2CH(3)OH contain only the syn isomer (S-CH(3) bond on the side of the Re=O bond), whereas ReOCl(2)(d-met) and ReOCl(2)(d,l-met) consist of the pure anti isomer. (1)H NMR spectroscopy shows that both isomers coexist in equilibrium in acetone (anti/syn ratio = 1:1 for X = Br, 3:1 for X = Cl). Exchange between these two isomers is fast above room temperature, but it slows down below 0 degrees C, and the sharp second-order spectra of both isomers at -20 degrees C were fully assigned. The coupling constants are consistent with the solid-state conformations being retained in solution. Complexes of the type [ReOX(2)(His-aa)]X (X = Cl, Br) are isolated with the dipeptides His-aa (aa = Gly, Ala, Leu, and Phe). X-ray diffraction work on [ReOBr(2)(His-Ala)]Br reveals the presence of distorted octahedral cations containing the Re=O(3+) core and a dipeptide coordinated through the histidine residue via the imidazole nitrogen, the terminal amino group, and the amide oxygen, the site trans to the Re=O bond being occupied by the oxygen. The alanine residue is ended by a protonated carboxylic group that does not participate in the coordination. The constant pattern of the(1)H NMR signals for the protons in the histidine residue confirms that the various dipeptides adopt a similar binding mode, consistent with the solid-state structure being retained in CD(3)OD solution. PMID- 15530098 TI - Considering Fe(II/IV) redox processes as mechanistically relevant to the catalytic hydrogenation of olefins by [PhBP iPr 3]Fe-H x species. AB - Several coordinatively unsaturated pseudotetrahedral iron(II) precursors, [PhBP(iPr)(3)]Fe-R ([PhBP(iPr)(3)] = [PhB(CH(2)P(i)Pr(2))(3)](-); R = Me (2), R = CH(2)Ph (3), R = CH(2)CMe(3) (4)) have been prepared from [PhBP(iPr)(3)]FeCl (1) that serve as precatalysts for the room-temperature hydrogenation of unsaturated hydrocarbons (e.g., ethylene, styrene, 2-pentyne) under atmospheric H(2) pressure. The solid-state crystal structures of 2 and 3 are presented. To gain mechanistic insight into the nature of these hydrogenation reactions, a number of [PhBP(iPr)(3)]-supported iron hydrides were prepared and studied. Room temperature hydrogenation of alkyls 2-4 in the presence of a trapping phosphine ligand affords the iron(IV) trihydride species [PhBP(iPr)(3)]Fe(H)(3)(PR(3)) (PR(3) = PMe(3) (5); PR(3) = PEt(3) (6); PR(3) = PMePh(2) (7)). These spectroscopically well-defined trihydrides undergo hydrogen loss to varying degrees in solution, and for the case of 7, this process leads to the structurally identified Fe(II) hydride product [PhBP(iPr)(3)]Fe(H)(PMePh(2)) (9). Attempts to prepare 9 by addition of LiEt(3)BH to 1 instead lead to the Fe(I) reduction product [PhBP(iPr)(3)]Fe(PMePh(2)) (10). The independent preparations of the Fe(II) monohydride complex [PhBP(iPr)(3)]Fe(II)(H)(PMe(3)) (11) and the Fe(I) phosphine adduct [PhBP(iPr)(3)]Fe(PMe(3)) (8) are described. The solid state crystal structures of trihydride 5, monohydride 11, and 8 are compared and demonstrate relatively little structural reorganization with respect to the P(3)Fe-P' core motif as a function of the iron center's formal oxidation state. Although paramagnetic 11 (S = 1) is quantitatively converted to the diamagnetic trihydride 5 under H(2), the Fe(I) complex 8 (S = (3)/(2)) is inert toward atmospheric H(2). Complex 10 is likewise inert toward H(2). Trihydrides 5 and 6 also serve as hydrogenation precatalysts, albeit at slower rates than that for the benzyl complex 3 because of a rate-contributing phosphine dependence. That these hydrogenations appear to proceed via well-defined olefin insertion steps into an Fe-H linkage is indicated by the reaction between trihydride 5 and ethylene, which cleanly produces the ethyl complex [PhBP(iPr)(3)]Fe(CH(2)CH(3)) (13) and an equivalent of ethane. Mechanistic issues concerning the overall reaction are described. PMID- 15530099 TI - Selenium and sulfur trichalcogenides from the chalcogenide exchange reaction. AB - Various selenium- and sulfur-containing volatiles have been detected and characterized as products of chalcogen exchange reactions. The formation of all possible selenium- and sulfur-containing trichalcogenide isomers (-SeSS-, -SSeS-, -SeSeS-, -SeSSe- and -SeSeSe-) was observed at room temperature in the solutions containing diselenide and trisulfide. Because of the low activation barrier of the selenium exchange reaction, these species are expected also to form in biological systems in the presence of diselenides and higher sulfides. Methyl and ethyl derivatives of these species were characterized using gas chromatography coupled to time-of-flight mass spectrometry with electron impact, chemical, and field ionization. Rearrangements of triselenides lead also to the formation of isomeric branched-structure triselenides, selanadiselenides. Because of the very similar structural properties, the behavior of these novel species was studied under various ionization modes (EI+, CI+, EI-, and CI-). PMID- 15530100 TI - Syntheses, characterization, and luminescence of Pt II-M I (M = Cu, Ag, Au) heterometallic complexes by incorporating Pt(diimine)(dithiolate) with [M2(dppm)2]2+ (dppm = Bis(diphenylphosphino)methane). AB - Reaction of Pt(diimine)(edt) (edt = 1,2-ethanedithiolate) with M(2)(dppm)(2)(MeCN)(2)(2+) (dppm = bis(diphenylphosphino)methane) gave heterotrinuclear complexes [PtCu(2)(edt)(mu-SH)(dppm)(3)](ClO(4)) (11) and [PtCu(2)(diimine)(2)(edt)(dppm)(2)](ClO(4))(2) (diimine = 2,2'-bpyridine (bpy), 12; 4,4'-dibutyl-2,2'-bipyridine (dbbpy), 13; phenanthroline (phen), 14; 5 bromophenanthroline (brphen), 15) when M = Cu(I). The reaction, however, afforded tetra- and trinuclear complexes [Pt(2)Ag(2)(edt)(2)(dppm)(2)](SbF(6))(2) (17) and [PtAu(2)(edt)(dppm)(2)](SbF(6))(2) (21) when M = Ag(I) and Au(I), respectively. The complexes were characterized by elemental analyses, electrospray mass spectroscopy, (1)H and (31)P NMR, IR, and UV-vis spectrometry, and X-ray crystallography for 14, 17, and 18. The Pt(II)Cu(I)(2) heterotrinuclear complexes 11-15 exhibit photoluminescence in the solid states at 298 K and in the frozen acetonitrile glasses at 77 K. It is likely that the emission originates from a ligand-to-metal charge transfer (dithiolate-to-Pt) (3)[p(S) --> d(Pt)] transition for 11 and from an admixture of (3)[d(Cu)/p(S)-pi(diimine)] transitions for 12 16. The Pt(II)(2)Ag(I)(2) heterotetranuclear complexes 17 and 18 are nonemissive in the solid states and in solutions at 298 K but show photoluminescence at 77 K. The Pt(II)Au(I)(2) heterotrinuclear complexes 19-21, however, are luminescent at room temperature in the solid state and in solution. Compounds 19 and 20 afford negative solvatochromism associated with a charge transfer from an orbital of a mixed metal/dithiolate character to a diimine pi orbital. PMID- 15530101 TI - Ylidene-->iminophosphine coordination complexes and reversible dissociation of dichlorophosphetidines. AB - Chloro-, bromo-, iodo-, and trifluoromethylsulfonyloxy-(2,4,6-tri-tert butylphenylimino)phosphines (MesNPX; X = Cl, Br, I, OTf) react quantitatively with 1,3-diisopropyl-4,5-dimethylimidazol-2-ylidene (Im) to give Lewis acid-base complexes with the general formula MesNP(Im)X. The dichlorophosphetidine (DippNPCl)(2) (Dipp = 2,6-diisopropylphenyl) represents a formal cyclodimer of an iminophosphine and reacts with Im to give a similar complex. The process represents a ligand induced dissociation of the phosphetidine framework and is reversed by the introduction of an appropriate Lewis acid. Solid state structures of RNP(Im)X complexes show that the closest contact between acid and base occurs between phosphorus and carbon in all cases, highlighting them as compounds that contain examples of C-->P coordinate bonds. Association of Im with phosphorus also effects a substantial increase in the P-X distance, but all derivatives maintain a short NP bond, indicating the presence of NP pi-bonding. PMID- 15530102 TI - Effects of Pt...Pt bonding, anions, solvate molecules, and hydrogen bonding on the self-association of Chugaev's cation, a platinum complex with a chelating carbene ligand. AB - Five salts, [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)](BPh(4)).CH(3)OH, [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)](PF(6)).CH(2)Cl(2), [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Cl.4H(2)O, [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Br.3.5H(2)O, and [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Cl.0.1H(2)O, have been crystallized and examined by single crystal X-ray diffraction. While the internal structure of the cation is similar in all salts, the interactions between cations vary in the different salts. Yellow [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)](BPh(4)).CH(3)OH and red [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)](PF(6)) form face-to-face dimers with Pt...Pt separations of 3.6617(6) and 3.340(2) A, respectively. In the latter, hydrogen bonding of the chelating ligand to adjacent anions facilitates the close approach of pairs of cations. The salts [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Cl.4H(2)O, [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Br.3.5H(2)O, and [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Cl.0.1H(2)O form columnar structures with Pt...Pt separations that range from 3.2514(5) to 3.5643(6) A. The water molecules and anions surround these columns and form bridges between neighboring columns. The electronic spectra of aqueous solutions of [(C(4)H(9)N(4))Pt(II)(CNCH(3))(2)]Cl.4H(2)O show spectral changes upon increasing concentrations of the platinum complex that are indicative of the formation of a dimer in solution with an equilibrium constant for dimerization of 23(1). PMID- 15530103 TI - (Fluoren-9-ylidene)methanedithiolato complexes of gold: synthesis, luminescence, and charge-transfer adducts. AB - Piperidinium 9H-fluorene-9-carbodithioate and its 2,7-di-tert-butyl-substituted analogue [(pipH)(S(2)CCH(C(12)H(6)R(2)-2,7)), R = H (1a), t-Bu (1b)] and 2,7 bis(octyloxy)-9H-fluorene-9-carbodithioic acid [HS(2)CCH(C(12)H(6)(OC(8)H(17))(2) 2,7), 2] and its tautomer [2,7-bis(octyloxy)fluoren-9-ylidene]methanedithiol [(HS)(2)C=C(C(12)H(6)(OC(8)H(17))(2)-2,7), 3] were employed for the preparation of gold complexes with the (fluoren-9-ylidene)methanedithiolato ligand and its substituted analogues. The gold(I) compounds Q(2)[Au(2)(mu-kappa(2)-S,S S(2)C=C(C(12)H(6)R(2)-2,7))(2)], where Q(+) = PPN(+) or Pr(4)N(+) for R = H (Q(2)4a) or Q(+) = Pr(4)N(+) for R = OC(8)H(17) [(Pr(4)N)(2)4c], were synthesized by reacting Q[AuCl(2)] with 1a or 2 (1:1) and excess piperidine or diethylamine. Complexes of the type [(Au(PR'3))(2)(mu-kappa(2)-S,S-S(2)C=C(C(12)H(6)R(2) 2,7))(2)] with R = H and R' = Me (5a), Et (5b), Ph (5c), and Cy (5d) or R = t-Bu and R' = Me (5e), Et (5f), Ph (5g), and Cy (5h) were obtained by reacting [AuCl(PR'(3))] with 1a,b (1:2) and piperidine. The reactions of 1a,b or 2 with Q[AuCl(4)] (2:1) and piperidine or diethylamine gave Q[Au(kappa(2)-S,S S(2)C=C(C(12)H(6)R(2)-2,7))(2)] with Q(+) = PPN(+) for R = H [(PPN)6a], Q(+) = PPN(+) or Bu(4)N(+) for R = t-Bu (Q6b), and Q(+) = Bu(4)N(+) for R = OC(8)H(17) [(Bu(4)N)6c]. Complexes Q6a-c reacted with excess triflic acid to give [Au(kappa(2)-S,S-S(2)C=C(C(12)H(6)R(2)-2,7))(kappa(2)-S,S-S(2)CCH(C(12)H(6)R(2) 2,7))] [R = H (7a), t-Bu (7b), OC(8)H(17) (7c)]. By reaction of (Bu(4)N)6b with PhICl(2) (1:1) the complex Bu(4)N[AuCl(2)(kappa(2)-S,S-S(2)C=C(C(12)H(6)(t-Bu)(2) 2,7))] [(Bu(4)N)8b] was obtained. The dithioato complexes [Au(SC(S)CH(C(12)H(8)))(PCy(3))] (9) and [Au(n)(S(2)CCH(C(12)H(8)))(n)] (10) were obtained from the reactions of 1a with [AuCl(PCy(3))] or [AuCl(SMe(2))], respectively (1:1), in the absence of a base. Charge-transfer adducts of general composition Q[Au(kappa(2)-S,S-S(2)C=C(C(12)H(6)R(2)-2,7))(2)].1.5TCNQ.xCH(2)Cl(2) [Q(+) = PPN(+), R = H, x = 0 (11a); Q(+) = PPN(+), R = t-Bu, x = 2 (11b); Q(+) = Bu(4)N(+), R = OC(8)H(17), x = 0 (11c)] were obtained from Q6a-c and TCNQ (1:2). The crystal structures of 5c.THF, 5e.(2)/(3)CH(2)Cl(2), 5g.CH(2)Cl(2), (PPN)6a.2Me(2)CO, and 11b were solved by X-ray diffraction studies. All the gold(I) complexes here described are photoluminescent at 77 K, and their emissions can be generally ascribed to LMMCT (Q(2)4a,c, 5a-h, 10) or LMCT (9) excited states. PMID- 15530104 TI - Bridged di(alkyl- and 4,4,4-trifluorobutylimidazolium) quaternary salts based on p-tert-butylcalix[4]arene. AB - Di(alkyl- and 4,4,4-trifluorobutylimidazolium) quaternary halides of p-tert butylcalix[4]arene, 5a-8a and 11a, were prepared and characterized. Metathetical reactions of these compounds with LiN(SO(2)CF(3))(2) and KPF(6) in methanol and water gave the corresponding new quaternary salts 5b-8b and 11b and 5c-8c and 11c, respectively, with high thermal stabilities. Compounds 5a-d, 6b, and 11a-c exhibit melting points <100 degrees C. On the basis of (1)H NMR and (13)C NMR spectral measurements, all of the new quaternary calix[4]arene salts were found to exist in a cone conformation. (1)H NMR titration experiments and electrospray MS spectra support the encapsulation of K(+) in the ionophoric cavity of 11c. PMID- 15530105 TI - Synthesis, structure, spectroscopic properties, and electrochemistry of (1,8,15,22-tetrasubstituted phthalocyaninato)lead complexes. AB - Three (1,8,15,22-tetrasubstituted phthalocyaninato)lead complexes Pb[Pc(alpha OR)(4)] [H(2)Pc(alpha-OC(5)H(11))(4) = 1,8,15,22-tetrakis(3 pentyloxy)phthalocyanine; H(2)Pc(alpha-OC(7)H(15))(4) = 1,8,15,22-tetrakis(2,4 dimethyl-3-pentyloxy)phthalocyanine; H(2)Pc(alpha-OC(10)H(7))(4) = 1,8,15,22 tetrakis(2-naphthyloxy)phthalocyanine] (1-3) have been prepared as racemic mixtures by treating the corresponding metal-free phthalocyanines H(2)Pc(alpha OR)(4) (4-6) with Pb(OAc)(2).3H(2)O in refluxing n-pentanol. The molecular structure of Pb[Pc(alpha-OC(5)H(11))(4)] (1) in the solid state has been determined by single-crystal X-ray diffraction analysis. This compound, having a nonplanar structure, crystallizes in the monoclinic system with a P2(1)/c space group. Each unit cell contains two pairs of enantiomeric molecules, which are linked by weak coordination of the Pb atom of one molecule with an aza nitrogen atom and its neighboring oxygen atom from the alkoxy substituent of another molecule, forming a pseudo-double-decker supramolecular structure in the crystals with a short ring-to-ring separation, 2.726 A, and thus a strong ring-ring pi-pi interaction. The decreased molecular symmetry for these complexes has also been revealed by the NMR spectra of 1 and 2. The methyl protons of the 3-pentyloxy and 2,4-dimethyl-3-pentyloxy side chains of 1 and 2, respectively, are chemically inequivalent. In addition to the elemental analysis and various spectroscopic characterizations, these compounds have also been electrochemically studied. Two one-electron oxidations and up to five one-electron reductions have been revealed by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methods. PMID- 15530106 TI - Chlorine dioxide reduction by aqueous iron(II) through outer-sphere and inner sphere electron-transfer pathways. AB - The reduction of ClO(2) to ClO(2)(-) by aqueous iron(II) in 0.5 M HClO(4) proceeds by both outer-sphere (86%) and inner-sphere (14%) electron-transfer pathways. The second-order rate constant for the outer-sphere reaction is 1.3 x 10(6) M(-1) s(-1). The inner-sphere electron-transfer reaction takes place via the formation of FeClO(2)(2+) that is observed as an intermediate. The rate constant for the inner-sphere path (2.0 x 10(5) M(-1) s(-1)) is controlled by ClO(2) substitution of a coordinated water to give an inner-sphere complex between ClO(2) and Fe(II) that very rapidly transfers an electron to give (Fe(III)(ClO(2)(-))(H(2)O)(5)(2+))(IS). The composite activation parameters for the ClO(2)/Fe(aq)(2+) reaction (inner-sphere + outer-sphere) are the following: DeltaH(r)++ = 40 kJ mol(-1); DeltaS(r)++ = 1.7 J mol(-1) K(-1). The Fe(III)ClO(2)(2+) inner-sphere complex dissociates to give Fe(aq)(3+) and ClO(2)( ) (39.3 s(-1)). The activation parameters for the dissociation of this complex are the following: DeltaH(d)++= 76 kJ mol(-1); DeltaS(d)++= 32 J K(-1) mol(-1). The reaction of Fe(aq)(2+) with ClO(2)(-) is first order in each species with a second-order rate constant of k(ClO2)- = 2.0 x 10(3) M(-1) s(-1) that is five times larger than the rate constant for the Fe(aq)(2+) reaction with HClO(2) in H(2)SO(4) medium ([H(+)] = 0.01-0.13 M). The composite activation parameters for the Fe(aq)(2+)/Cl(III) reaction in H(2)SO(4) are DeltaH(Cl(III))++ = 41 kJ mol( 1) and DeltaS(Cl(III))++ = 48 J mol(-1) K(-1). PMID- 15530107 TI - Synthesis and characterization of bismuth single-crystalline nanowires and nanospheres. AB - A facile solution-phase process has been demonstrated for the selective preparation of single-crystalline bismuth nanowires and nanospheres by reducing sodium bismuthate with ethylene glycol in the presence of poly(vinyl pyrrolidone) (PVP) or acetone. Bismuth nanobelts and Bi/Bi(2)O(3) nanocables could be also obtained by changing some reaction parameters. Various techniques such as XRD, EDXA, SEM, TEM, HRTEM, and FT-IR have been used to investigate the physical characteristics of these low-dimensional bismuth nanostructures. PMID- 15530108 TI - An efficient and improved route for the preparation of (S)-5 aminomethyloxazolidinone libraries. PMID- 15530109 TI - Traceless solid-phase synthesis of carbolinones. PMID- 15530110 TI - A sensitive colorimetric method for the study of polystyrene merrifield resins and chloromethylated macroporous monolithic polymers. PMID- 15530111 TI - Microarrays of tagged combinatorial triazine libraries in the discovery of small molecule ligands of human IgG. AB - A novel and highly diverse tagged triazine library incorporating a triethylene glycol-based linker was synthesized using an orthogonal combinatorial approach on the solid phase and covalently immobilized on a glass substrate as a small molecule microarray (SMM). The SMM was screened with a fluorophore-conjugated human IgG, and 4 novel binders from a library of 2688 compounds were identified from the fully spatially addressable array without the need for compound decoding. Using surface plasmon resonance (SPR) analysis, binding seen on the array was confirmed, and a binding constant as low as Kd = 2.02 x10(-6) M was measured. PMID- 15530112 TI - Resonant multisensor system for high-throughput determinations of solvent/polymer interactions. AB - Solvent-resistant polymers are important in numerous research, engineering, and consumer applications. To address the limitations of existing methods of evaluation of polymer solubility and solvent resistance, we developed and built a 6 x 4 array of resonant acoustic-wave sensors operating in the thickness shear mode (TSM). The application of this system makes possible analysis of nanogram quantities of polymers in small amounts of solvent and permits the simultaneous analysis of multiple samples, such as those produced in combinatorial polymerization reactions. These parallel determinations of polymer/solvent interactions eliminate errors associated with serial determinations. During the periodic exposure of the TSM crystals to polymer/solvent combinations, the mass increase of the crystal is determined, which is proportional to the amount of polymer dissolved and deposited onto the sensor from a polymer solution. We demonstrate our sensor system for reliable quantification of solubility of several types of polymers in various solvents. The high mass sensitivity of our resonant TSM sensors (10 ng), use of only a minute volume of a solvent (< 2 mL), and parallel operation (matching a layout of available 24 well plates) make this system a good fit with available polymer combinatorial synthesis equipment. PMID- 15530113 TI - Preparative LC-MS purification: improved compound-specific method optimization. AB - One of the remaining challenges in providing effective preparative LC-MS purification is balancing throughput and compound purity. We describe here an approach to optimizing preparative LC-MS methods that provides significantly better chromatographic resolution and, hence, better compound purity than generic preparative LC methods consuming the same amount of time. This approach is easier to implement, is more rugged, and permits significantly greater flexibility than previously reported approaches. The instrument configurations and protocols presented here are specifically tailored for open access support, but the basic approach is equally suitable and effective in high-throughput situations. PMID- 15530114 TI - Combining Biginelli multicomponent and click chemistry: generation of 6-(1,2,3 triazol-1-yl)-dihydropyrimidone libraries. AB - Efficient solution-phase protocols for the high-throughput synthesis of 6-(1,2,3 triazol-1-yl)-dihydropyrimidones are reported. The multistep sequence involves the initial bromination of dihydropyrimidones precursors (DHPMs, Biginelli compounds) at the C6-methyl position, using a recyclable polymer-supported brominating agent under rapid flow-through conditions (residence time of 1 min). The 6-bromomethyldihydropyrimidone intermediates were subsequently subjected to a microwave-assisted azidation step (25 min), providing the key 6 azidomethyldihydropyrimidone precursors. In the final step of the sequence, the azides were treated with terminal acetylenes under Cu(I) catalysis (azide acetylene ligation, "click chemistry") to provide the target 6-(1,2,3-triazol-1 yl)-dihydropyrimidones in a regiospecific fashion (1,4-triazoles) in moderate overall yield utilizing controlled microwave irradiation (20 min). In total, a library of 27 compounds was prepared with 4 points of diversity. PMID- 15530116 TI - Traceless solid-phase synthesis of substituted benzimidazolones. PMID- 15530115 TI - Three-step solution-phase combinatorial access to 1,2-disubstituted and 1,2,5 trisubstituted pyrroles from carboxylic esters. AB - An efficient diversity-oriented strategy has been developed for the solution phase parallel synthesis of di- and trisubstituted pyrrole libraries. Methyl esters 1 were effectively converted to 1,2-di- and 1,2,5-trisubstituted pyrroles 5 and 6 in three steps. Treatment of ester 1 with vinylmagnesium bromide in the presence of copper (I) cyanide yielded the corresponding homoallylic ketone 2, which was subjected to ozonolysis or Tsuji-Wacker oxidation to yield the respective cyclization precursors 3 and 4 after aqueous workup. Compounds 3 and 4 were condensed without further purification with a primary amine to afford the desired 1,2-di- or 1,2,5-trisubstituted pyrroles 5 and 6 in good yield and purity. PMID- 15530117 TI - Solid-phase synthesis of a 4-substituted gamma-lactam library. AB - Pyrrolidin-2-one (gamma-lactam) derivatives have shown a wide range of activities as ligands to diverse receptors, e.g., integrin, CCR5, and CCK. Therefore, we have prepared a large library of these derivatives for high-throughput screening against various protein targets, after developing a synthesis of pyrrolidin-2 ones on solid phase. Exploration of the ring formation was key to the success of this synthesis. First, acylation of resin-bound amines with N-Fmoc-protected amino acids and subsequent deprotection of the Fmoc group were accomplished readily. The resulting resin-bound primary amines were treated with beta monomethyl itaconate under gentle heat in a mixture of methanol and toluene to yield the desired intermediates. Finally, saponification, amide formation, and cleavage from the resin led to the production of a library of 12,000 pyrrolidin-2 one derivatives. These products were isolated as diastereomeric mixtures of high purity and were obtained in good yields. PMID- 15530118 TI - General inverse solid-phase synthesis method for C-terminally modified peptide mimetics. AB - Peptide mimetics are of considerable interest as bioactive agents and drugs. C terminally modified peptide mimetics are of particular interest given the synthetic versatility of the carboxyl group and its derivatives. A general approach to C-terminally modified peptide mimetics, based on a urethane attachment strategy and amino acid t-butyl ester-based N-to-C peptide synthesis, is described. This approach is compatible with the reaction conditions generally employed for solution-phase peptide mimetic synthesis. To develop and demonstrate this approach, it was employed for the solid-phase synthesis of peptide trifluoromethyl ketones, peptide boronic acids, and peptide hydroxamic acids. The development of a versatile general approach to C-terminally modified peptides using readily available starting materials provides a basis for the combinatorial and parallel solid-phase synthesis of these peptide mimetic classes for bioactive agent screening and also provides a basis for the further development of solid phase C-terminal functional group elaboration strategies. PMID- 15530119 TI - Prediction of HPLC conditions using QSPR techniques: an effective tool to improve combinatorial library design. AB - The purification and characterization of compounds resulting from parallel synthesis or combinatorial chemistry has not yet been optimized to operate as a completely automated high-throughput process. Liquid chromatography/mass spectroscopy (LC/MS) is most commonly employed to carry out the characterization and identification of combinatorial compounds. This desired level of automation can only be accomplished if the separation conditions for every compound in the combinatorial array are known prior to the analysis. This study presents a quantitative structure retention relationship (QSRR) approach to predict the retention time of structurally diverse solutes under 75 different LC/MS conditions. Sixty-two compounds were analyzed using 15 commonly used HPLC columns under 5 different gradient conditions. The solute retention time was used as the dependent variable, and more than 1000 molecular descriptors were calculated for this compound set to generate QSRR models. After the elimination of highly correlated variables and those with zero variance, two different genetic algorithms were applied to identify the most significant descriptors. Following the variable selection, the identified descriptors were used to create QSRR models for each separation condition. The calculated stepwise multiple linear regression models have been proven to be statistically significant and highly predictive, with an average coefficient of determination (R2) of 0.86, an average cross-validated r2 of 0.62, r2 = 0.76, and an average F value of 27.29. The QSRR models can be used to design "analysis-friendly" library purification plates, in which compounds are arranged on the basis of their predicted separation condition and can also be used during the library design phase to flag compounds not amenable to the separation methods in use. PMID- 15530120 TI - Traceless solid-phase synthesis of nitrogen-containing heterocycles and their biological evaluations as inhibitors of neuronal sodium channels. AB - The preparation of pyrimidine-2-thione, pyrimidine-2-one, pyrimidine, and benzo[b][1,4]diazepine derivatives using traceless solid-phase sulfone linker strategy is described. Key steps involved are (i) sulfinate S-alkylation, (ii) sulfone anion alkylation with an epoxide, (iii) gamma-hydroxyl sulfone --> gamma ketosulfone oxidation, and (iv) traceless product release by a one-pot elimination-cyclization process. Elimination-cyclization was carried out under basic conditions with thiourea, methyl thiourea, methyl urea, guanidine hydrochloride, benzamidine hydrochloride and ortho-phenylene diamine. Twenty three compounds were prepared, and 14 of them were evaluated by the Batrachotoxin (BTX) radioligand binding assay for their binding affinity to neuronal sodium channels. Compound 7c was found to be a potential neuronal sodium channels blocker. PMID- 15530121 TI - Solid-phase synthesis of N-bi- and N-terarylpiperazines via three different and consecutive palladium-catalyzed cross-coupling reactions. AB - Novel nonsymmetrical N-bi- and N-terarylpiperazines were synthesized on solid phase via three consecutive palladium-catalyzed cross-coupling reactions in which C-N, C-B, and C-C bonds are formed consecutively. PMID- 15530122 TI - Solution-phase parallel synthesis of an N-alkylated dihydropteridinone library from fluorous amino acids. AB - Parallel synthesis of an N-alkylated dihydropteridinone library has been accomplished in five steps starting from two displacement reactions of 4,6 dichloro-5-nitropyrimidine, first with fluorous amino acids, then with secondary amines. The hydrogenation of the nitro group followed by microwave-assisted cyclization gave the dihydropteridinones. Further diversification was achieved by the reaction of dihydropteridinones with benzyl halides to afford mono-N alkylated products. All the reaction intermediates and final products were purified by SPE or precipitation without the need to perform chromatography. PMID- 15530123 TI - A base-catalyzed solution-phase parallel synthesis of substituted vinylic benzamides from 3-amino-2-cyclohexanones. AB - An improved method for the synthesis of benzamides from 3-amino-2-cyclohexenones is presented. Using sodium hydride, a base-catalyzed N-benzoylation provided significantly higher yields (71-79%) for the reported compounds. This novel protocol was applied in the solution-phase parallel synthesis of a 12-member library of vinylic benzamide derivatives of 3-amino-2-cyclohexenones in 63-90% yield, using a Radley's Carousel Reaction Station. PMID- 15530124 TI - Polystyrene-supported triphenylarsine reagents and their use in Suzuki cross coupling reactions. AB - Soluble and insoluble polystyrene-bound triphenylarsine reagents have been prepared from 4-styryldiphenylarsine. The utility of these reagents as ligands for palladium in Suzuki cross-coupling reactions is demonstrated. In these applications, the use of polymeric triphenylarsine simplifies the purification of the coupled product and allows for the ligand to be recycled. Furthermore, the soluble polymeric arsine reagent permits the palladium catalyst to be recovered and reused. PMID- 15530125 TI - Functionalized cross-linked poly(vinyl alcohol) resins as reaction scavengers and as supports for solid-phase organic synthesis. AB - New hydrophilic poly(vinyl alcohol) (PVA-OH) resins were prepared by an inverse suspension polymerization using epichlorohydrin as a cross-linker. These novel resins swell in a variety of solvents commonly used in solid-phase organic synthesis, such as dicholomethane, dioxane, methanol, tetrahydrofuran, and dimethylformamide. In addition, PVA-OH shows excellent swelling in water. The cross-linked PVA-OH beads were functionalized with an aldehyde group and were tested as scavengers for primary amines in three different reactions: amide bond formation, reductive amination reaction, and urea formation. With 1-2 equiv of the PVA aldehyde resin, all the excess primary amines were successfully scavenged. The utility of PVA-OH resins as solid supports in mono- and dipeptide synthesis was also investigated using symmetrical anhydride and MSNT/MeIm (2,4,6 mesitylenesulfonyl-3-nitro-1,2,4-triazolide in the presence of 1-methylimidazol) methods. PMID- 15530126 TI - Parallel screening of homogeneous copper catalysts for the oxidation of benzylic alcohols with molecular oxygen in aqueous solutions. AB - A simple and efficient parallel screening method to evaluate the catalytic activities of homogeneous copper complexes for the oxidation of benzylic alcohols in aqueous solutions with molecular oxygen is reported. Copper(II) sulfate was treated in situ with 22 nitrogen donor ligands, and the catalytic activities of these combinations were studied at four different pH values with two substrates (benzyl alcohol and 3,4-dimethoxy benzyl alcohol (veratryl alcohol)), resulting in 176 oxidation experiments in the primary screening stage. Copper complexes based on N,N,N',N'-tetramethyl ethylenediamine (TMEDA), 9,10-diaminephenanthrene (DAPHEN), and 1,2-diaminocyclohexane (DACH) were determined to be the most active catalysts. In the second screenings, the influence of reaction conditions on Cu(DACH)-, Cu(TMEDA)-, and Cu(DAPHEN)-catalyzed reactions were investigated in more detail. It was found that highly basic reaction conditions favor the reaction with the exception of Cu(TMEDA), which is active at a lower pH range. Under optimized conditions, Cu(DAPHEN) catalyzes the transformation of veratryl alcohol to the corresponding aldehyde with 100% conversion. PMID- 15530127 TI - Enteral drug absorption in patients with short small bowel : a review. AB - Drug therapy may become difficult when a significant amount of the small intestine is resected, as happens in patients with a short small bowel. Drug absorption from the gastrointestinal tract is altered in these patients; however, this effect is variable in patients and differs with each drug. Literature regarding clinical outcomes of normal or alternative administration routes in patients with a short small bowel is limited. We explored what is written about the normal absorption of commonly used drugs and what difference the resection of different but substantial parts of the small intestine makes. Changes in the gastrointestinal tract after resection of >50% of the small intestine causes malabsorption of macronutrients and micronutrients, and may alter the drug absorption process. The metabolic activity of the abundantly present intestinal lactobacilli can also affect the enteral drug absorption in patients with short small bowel as this results in the production of lactic acid, gaseous CO(2), ethanol and an increased bile acid deconjugation. Accelerated intestinal luminal transit time causes a reduction in absorption of certain antimicrobial agents, digoxin, hydrochlorothiazide, ciclosporin, cimetidine, mesalazine (5 aminosalicylic acid), oral contraceptives and levothyroxine. Gastric hypersecretion and lack of sufficient contact time with the intestinal mucosa in patients with short small bowel leads to insufficient absorption of drugs such as omeprazole. Successful treatment with warfarin, tricyclic antidepressants, metronidazole, fluconazole, procainamide, sotalol and pindolol are reported in several studies. Many different factors cause this variability in drug absorption in such patients. Monitoring the serum drug concentration in these patients may ease dealing with the management problems. PMID- 15530128 TI - Avoidance of bleeding during surgery in patients receiving anticoagulant and/or antiplatelet therapy: pharmacokinetic and pharmacodynamic considerations. AB - Perioperative management of chronically anticoagulated patients and/or patients treated with antiplatelet therapy is a complex medical problem. This review considers the pharmacokinetic and pharmacodynamic properties of commonly used antiplatelet and anticoagulant drugs with special emphasis on loss of effects after discontinuation and possible counteracting (or antidote) strategies. These drugs are aspirin (acetylsalicylic acid), ticlopidine/clopidogrel, abciximab, tirofiban and eptifibatide, heparin (unfractionated and low-molecular-weight), warfarin and direct thrombin inhibitors. Since the pharmacological mechanisms of some of these drugs are based on irreversible or slowly reversible effects, their pharmacokinetic profiles are not necessarily predictive for their pharmacodynamic profiles. A close and direct relationship between plasma concentrations and effects is seen only for the glycoprotein (GP) IIb/IIIa inhibitors tirofiban and eptifibatide with a fast off-rate for dissociation from the GPIIb/IIIa receptor, and for direct thrombin inhibitors (hirudin and argatroban). For other compounds, drug concentrations in plasma and pharmacodynamic effects are not closely correlated because of, for example, irreversible binding to their target (aspirin, clopidogrel and abciximab), inhibition of the generation of a subset of clotting factors with differing regeneration and degradation rates (coumarins) or sustained binding to the vascular wall (heparins). Surgery in patients on anticoagulant and/or antiplatelet therapy may be categorised as: (i) elective versus urgent; and (ii) cardiopulmonary bypass (CPB) versus non-CPB. Monotherapy with clopidogrel or aspirin need not be discontinued in elective non-CPB surgery, and temporary discontinuation of warfarin should be accompanied by preoperative intravenous heparin only in selected high-risk patients. Vitamin K as an antidote for warfarin should only be used subcutaneously and solely in urgent/emergency surgery. In elective surgery requiring CPB (coronary artery bypass grafting), it is recommended to discontinue aspirin 7 days preoperatively in patients with a low risk profile. Patients requiring urgent CPB surgery (e.g. after failure of a percutaneous coronary angioplasty with or without coronary stent deployment) are usually pretreated with several antiplatelet agents (e.g. aspirin and clopidogrel, together with a GPIIb/IIIa inhibitor) together with unfractionated or low-molecular-weight heparin. With judicious planning, urgent/emergency cardiac surgery can be safely performed on these patients. Delaying surgery (e.g. for 12 hours in patients treated with abciximab) should be considered if possible. Standard heparin doses should be given to achieve optimal anticoagulation for CPB. Prophylactic use of aprotinin (intra- and/or postoperatively), aminocaproic acid or tranexamic acid should be considered. Early (in the operating theatre prior to chest closure) and judicious use of replacement blood products (platelets) should be commenced when clinically indicated. PMID- 15530130 TI - Reduction of saquinavir exposure by coadministration of loperamide: a two-way pharmacokinetic interaction. AB - OBJECTIVE: To assess any pharmacokinetic interactions between loperamide and saquinavir. DESIGN: Double-blind, double-dummy, randomised, placebo-controlled, three-way crossover trial. PARTICIPANTS: Twelve healthy male and female volunteers, aged 24-46 years. METHODS: Saquinavir and loperamide pharmacokinetics were determined over a 72-hour period after single dose administration of saquinavir 600mg and/or loperamide 16mg. Plasma and urine concentrations of loperamide, its metabolites, and saquinavir were analysed using a single liquid chromatography/tandem mass spectrometry method for all compounds. RESULTS: Saquinavir exposure was reduced by 54% when given with loperamide (median area under the concentration-time curve from zero to infinity [range], 1189 [243-2113] vs 550 [234-1468] pmol . h/mL; p = 0.016) with unchanged renal clearance. In contrast, loperamide concentrations increased and desmethylloperamide concentrations decreased during saquinavir coadministration, resulting in a reduced metabolic clearance of loperamide (median [range], 544 [224-1393] vs 443 [238-692] mL/min; p = 0.016). CONCLUSIONS: Whereas the effect of saquinavir on loperamide disposition is unlikely to be of clinical relevance, the reduced drug exposure of saquinavir when loperamide is coadministered is worrisome because a relationship between protease inhibitor drug exposure and antiviral response has been reported. Patients receiving saquinavir monotherapy should be advised not to combine these drugs, especially for prolonged periods of time because a reduction in therapeutic efficacy may result. PMID- 15530131 TI - Pharmacokinetics of quetiapine in elderly patients with selected psychotic disorders. AB - OBJECTIVE: To assess the pharmacokinetics and tolerability of quetiapine in elderly patients with selected psychotic disorders. STUDY DESIGN: This was a multicentre, open-label, 27-day, rising multiple-dose trial. Descriptive statistics summarised plasma quetiapine concentrations and pharmacokinetic parameters by trial day. A two-way analysis of variance was used to evaluate all pharmacokinetic parameters, and 90% confidence intervals of the mean differences were calculated. METHODS: Antipsychotic drugs taken prior to the study period were discontinued on day 1. Quetiapine treatment began on day 3. Doses were increased stepwise, starting at 25mg three times daily and reaching 250mg three times daily by day 21. PATIENTS: Twelve patients (age 63-85 years) with schizophrenia, schizoaffective disorder or bipolar disorder. MAIN OUTCOME MEASURES AND RESULTS: Key assessments included quetiapine plasma concentrations, and neurological and safety evaluations. Under steady-state conditions, the 100 and 250mg doses of quetiapine were not significantly different in terms of dose normalised area under the plasma concentration-time curve within an 8-hour dose administration interval, or in dose-normalised minimum plasma concentration (C(min)) at the end of a dose administration interval. The morning C(min) values for the seven discrete dose amounts evaluated also increased linearly with dose. The apparent oral clearance, volume of distribution and half-life did not change as a function of dose. There were no serious adverse events. The most common adverse events were postural hypotension (n = 6), dizziness (n = 5) and somnolence (n = 4). CONCLUSIONS: While quetiapine was well tolerated at doses up to 250mg three times daily, the potential for reduced clearance, as well as the adverse effects of postural hypotension and dizziness, indicated that quetiapine should be introduced at lower doses and titrated at a relatively slower rate in patients > or =65 years. PMID- 15530132 TI - Assessment of levothyroxine sodium bioavailability: recommendations for an improved methodology based on the pooled analysis of eight identically designed trials with 396 drug exposures. AB - BACKGROUND: Assessment of dosage form performance in delivering endogenous compounds, such as hormones, in vivo requires a specific approach. OBJECTIVES: Assessment of relative bioavailability of levothyroxine sodium (L-T4) from eight solid preparations, compared with a liquid formulation, by using pharmacological doses, and critical evaluation of trial methodology based on the pooled analysis of individual data. DESIGN: Eight open-label, randomised, single-dose, crossover phase I studies using eight solid L-T4 dosage forms (25, 50, 75, 100, 125, 150, 175, 200 microg per tablet; administered total doses 600, 625 or 700 microg) and a liquid formulation; assessment of relative bioavailability by 90% confidence intervals for the relative area under the concentration-time curve (AUC) of total thyroxine (TT4), i.e. protein-bound plus free thyroxine, calculated by using the recommended log AUC four-way analysis of variance models for crossover designs. For the pooled analysis, general linear models were applied to assess the validity of model assumptions, to identify potential sources of effect modification, to discuss alternative modelling approaches with respect to endogenous hormone secretion and to give recommendations for future designs and sample sizes. PARTICIPANTS: One hundred and sixty-nine healthy males; 29 of these individuals participating in two studies. INTERVENTIONS: Single oral doses of L T4 tablets and the liquid formulation administered after fasting, separated by at least 6 weeks; a total of 396 drug exposures. MAIN OUTCOME MEASURES: TT4 AUC from 0 to 48 hours and peak plasma concentration with and without baseline correction. RESULTS: Each study demonstrated equivalence of the tablets to the drinking solution, independent of the chosen analysis model. Sequence effects that could devalidate the chosen crossover approach were not found. Period effects with changing directions that could best be explained by seasonal variation were detected. While the pre-specified method of baseline correction of simply subtracting individual time-zero TT4 values was disadvantageous, the analysis of total AUC could be improved considerably by covariate adjustment for baseline TT4. With this approach, sample sizes could have been substantially reduced or, alternatively, the recommended equivalence ranges could be reduced to +/-6%. CONCLUSION: Using a single pharmacological dose of L-T4 in two-period crossover designs is a safe and reliable procedure to assess L-T4 dosage form performance. With an adequate statistical modelling approach, the design is efficient and allows general conclusions with moderate sample sizes. PMID- 15530133 TI - Modelling ciclosporin double-peak absorption profiles in the early post transplantation period. AB - OBJECTIVE: To apply a recent double-peak absorption model to ciclosporin. METHODS: Pharmacokinetic evaluation was performed in 20 patients who had undergone de novo renal transplantation and were receiving immunosuppressive therapy with corticosteroids, mycophenolate mofetil and ciclosporin Neoral. Data were analysed by nonlinear mixed-effects modelling using individual and population approaches. RESULTS: Six patients presented pharmacokinetic profiles with two peaks, whereas 14 curves apparently had a single, sometimes delayed and wide, peak. A one-compartment model with first-order elimination in association with the double-peak absorption model best fitted the data. This model reliably described the pharmacokinetics of ciclosporin, whether or not a double peak was present on the concentration-time profiles. CONCLUSIONS: The double-peak absorption model could be useful to optimise ciclosporin exposure in the early post-transplantation period. PMID- 15530135 TI - Is the "ownership society" the solution to the ever-rising costs of medical care? Plus: correcting overcorrected esotropias and A-pattern anisotropias; oculocardiac reflex; myasthenia gravis; aniseikonia on the Hess test; contracture of the inferior oblique after cataract surgery. PMID- 15530129 TI - Genetic predictors of the clinical response to opioid analgesics: clinical utility and future perspectives. AB - This review uses a candidate gene approach to identify possible pharmacogenetic modulators of opioid therapy, and discusses these modulators together with demonstrated genetic causes for the variability in clinical effects of opioids. Genetically caused inactivity of cytochrome P450 (CYP) 2D6 renders codeine ineffective (lack of morphine formation), slightly decreases the efficacy of tramadol (lack of formation of the active O-desmethyl-tramadol) and slightly decreases the clearance of methadone. MDR1 mutations often demonstrate pharmacogenetic consequences, and since opioids are among the P-glycoprotein substrates, opioid pharmacology may be affected by MDR1 mutations. The single nucleotide polymorphism A118G of the mu opioid receptor gene has been associated with decreased potency of morphine and morphine-6-glucuronide, and with decreased analgesic effects and higher alfentanil dose demands in carriers of the mutated G118 allele. Genetic causes may also trigger or modify drug interactions, which in turn can alter the clinical response to opioid therapy. For example, by inhibiting CYP2D6, paroxetine increases the steady-state plasma concentrations of (R)-methadone in extensive but not in poor metabolisers of debrisoquine/sparteine. So far, the clinical consequences of the pharmacogenetics of opioids are limited to codeine, which should not be administered to poor metabolisers of debrisoquine/sparteine. Genetically precipitated drug interactions might render a standard opioid dose toxic and should, therefore, be taken into consideration. Mutations affecting opioid receptors and pain perception/processing are of interest for the study of opioid actions, but with modern practice of on-demand administration of opioids their utility may be limited to explaining why some patients need higher opioid doses; however, the adverse effects profile may be modified by these mutations. Nonetheless, at a limited level, pharmacogenetics can be expected to facilitate individualised opioid therapy. PMID- 15530136 TI - Outcome of a surgical treatment protocol for late consecutive exotropia following bilateral medial rectus recession for esotropia. AB - PURPOSE: To describe a specific protocol for the surgical treatment of Late Consecutive Exotropia (LCXT) following bilateral medial rectus (MR) recessions, and to report the results of surgery, performed according to this protocol, in a series of 119 children followed 6 months to 15 years after surgery for consecutive exotropia. METHOD: The senior author (JLM) performed a retrospective chart review of 15 years of his experience following a specific protocol for the surgical treatment of LCXT. In individual cases, following this protocol led to recessions of the overacting inferior oblique (IO) with anterior transposition if dissociated vertical deviation (DVD) was also present, weakening of the overacting superior oblique (SO) with simultaneous 10 mm recessions of the superior rectus (SR) with 3 mm of nasal transposition (to ameliorate DVD which might be aggravated by SO tenotomies), graded recessions of one lateral rectus (LR) 7.0 to 8.5 mm if the distance deviation was equal to or less than 23 prism diopters (PD) of XT, and advancement of one previously recessed MR to the insertion if the distance deviation was equal to or greater than 24 PD XT. RESULTS: "Success" was defined as 8 PD ET to 6 PD XT, with both distance and near fixation. Among 14 patients receiving SR recessions, SO tenotomies, and recession of one LR, 12 of the 14 were "successes" at 6 months postop' and 10 of 14 at an average of 3 years postop'. Among 26 receiving advancement of one MR, "successes" were 23 of 26 at 6 months postop' and 20/26 at two years after surgery. Among 22 receiving IO recession with anterior transposition if DVD was present, "successes" were 18 of 22 at 6 months and 12 of 22 at 3.6 years. Among 57 receiving only recession of one LR, 53 of 57 were "successes" at 6 months postop' and 45 of 57 at 4 years postop'. The overall "success" rate for this protocol for these 119 patients was 89% at 6 months and 74% at 2 to 4 years postop'. Treatment of simultaneous IO overaction and LCXT was associated with the highest long term "failure" rate in this protocol. CONCLUSION: This specific surgical protocol for treatment of late consecutive exotropia is reasonably safe and effective and comparable to the few other reported protocols of various surgical treatments for consecutive exotropia. PMID- 15530134 TI - Effect of antacid on the pharmacokinetics of extended-release formulations of tolterodine and oxybutynin. AB - BACKGROUND: In general, extended-release (ER) formulations are designed to prolong the duration of efficacy and reduce the adverse effects of a drug. These formulations often contain the entire daily dose in a single tablet. Therefore, failure of the ER mechanism not only diminishes the desired benefits, but may temporarily expose the patient to drug concentrations higher than those released from a conventional tablet. In this study we determined whether pH has an effect on drug release from the ER formulations of oxybutynin (OROS technology) and tolterodine (membrane coated beads) in vitro and in vivo. STUDY DESIGN: In vitro studies were based on standardised dissolution experiments for each drug in media of different pH (artificial gastric fluid at pH 1.2, artificial intestinal fluid at pH 7.5, and water). In the two separate, identically designed in vivo studies, single doses of each drug were administered alone and with an antacid to male and female healthy volunteers aged 18-45 years. The randomised, crossover, open-label in vivo studies employed a validated assay to determine plasma concentrations of tolterodine and its metabolite 5-hydroxymethyl tolterodine (5-HM), or oxybutynin and its metabolite N-desethyloxybutynin. RESULTS: The in vitro study showed similar slow and steady drug release from ER-oxybutynin in each pH medium, with 64-71% released after 12 hours. Drug release from ER-tolterodine was steady and slow in artificial gastric fluid, with 72.5% of drug released after 12 hours. However, drug release was much faster in artificial intestinal fluid and water, where 69.8% and 69.1%, respectively, of the drug was released within 4 hours. These in vitro results were consistent with the findings of the in vivo studies. In vivo, the pharmacokinetic profile (peak plasma concentration [C(max)] and area under the concentration-time curve) of ER-oxybutynin was similar after administration with or without antacid, whereas C(max) values of both tolterodine and 5-HM increased significantly when ER-tolterodine was administered with antacid (p < or = 0.017 vs ER-tolterodine alone). CONCLUSIONS: Changes in pH affected the release of tolterodine from ER-tolterodine, while they had no effect on the release of oxybutynin from the proprietary ER technology used in ER oxybutynin. The technology employed in ER formulations thus determines sensitivity of drug release to external factors. PMID- 15530137 TI - Vertical transposition of medial rectus muscles for correction of A-pattern anisotropia. AB - BACKGROUND AND PURPOSE: A-pattern esotropias without superior oblique muscle overaction form a small but distinct group among A and V pattern anisotropias. The purpose of this study was to determine the results of ungraded 5 mm bilateral medial rectus muscle transpositions as a treatment of A anisotropia in esotropic patients, and to study these results with relation to the magnitude of preoperative A anisotropia and to the magnitude of preoperative esotropia in primary position. METHODS: Patient data on all 37 esotropic patients who had 5 mm elevation of the medial rectus muscle insertion for correction of A anisotropia at our institution during the previous 25 years were studied. We considered the surgical result to be "satisfactory" when a total correction, and A pattern less than 10 Prism Diopters (PD) or a V pattern less than 15 PD were obtained. The patients were divided into different groups, according to the magnitude of the preoperative A pattern and according to the magnitude of the preoperative esotropia in primary position. For statistical analysis, the Chi Square and the Fisher Tests were employed. RESULTS: "Satisfactory" results were found in 70.3% of the cases. The mean correction of A anisotropia was 11.4 PD (76.1%), which corresponded to 2.3 PD per millimeter of transposition. "Unsatisfactory" results were more prevalent in patients with esotropia in primary position larger than 40 PD (p=0.0418). CONCLUSIONS: A five millimeter vertical transposition (elevation) of both medial rectus muscles is an effective treatment for correction of A anisotropia in most esotropic patients, but is associated with unsatisfactory results when the preoperative esotropia in primary position is larger than 40 PD. PMID- 15530138 TI - The profound augmentation of the oculocardiac reflex by fast acting opioids. AB - BACKGROUND: Trigemino-vagal bradycardia elicited by tension on an extraocular muscles (oculocardiac reflex; OCR) is a hazard for strabismus surgery patients. OCR can be reduced by anticholinergic medications and regulating the depth of anesthesia. We investigated the influence of narcotic agents as a routine part of general anesthesia for strabismus surgery in adults and children. METHOD: From August, 1992 through September, 2000, 1275 patients undergoing extraocular muscle surgery were prospectively studied during 10 second, 200 gram square wave tension on gently isolated rectus muscles. The anesthetic agents, gas concentrations, and patient age were recorded. Patients receiving an anticholinergic agent and reoperated cases were excluded, yielding 1029 study cases. 849 received no narcotic as a part of induction. Two groups of patients were given no opiate before the first inferior rectus muscle was pulled. A narcotic was administered IV 5 minutes before the second inferior rectus was pulled. One group of 49 received fentanyl 0.15 uL/Kg and the second group of 12 received meperidine 1 mg/kg. RESULTS: Faster acting intravenous induction opioids had a profound augmenting effect on the degree of oculocardiac reflex. Compared to no narcotic, the OCR was increased most by remifentanyl (p less than .0001), then sufenta (p=.02), and fentanyl (p less than .0001). Induction morphine had no appreciable effect on the OCR (p=.9). For the 49 patients with IV fentanyl delivered between the first OCR and second OCR, a significant increase occurred (p=.003). This increase in %OCR was not correlated with a change in inhalational gas concentration (p=.9), CO2 concentration (p=.6) or age (p=.12). For the 12 patients given demerol between the first and second rectus tension, no significant OCR change occurred (p=.7). The OCR was greatest for inferior rectus, then superior rectus, then medial rectus, and least for lateral rectus. (See text for details.) CONCLUSION: In the absence of anticholinergic blockade, rapidly acting narcotics enhance the degree of bradycardia due to the OCR elicited by controlled extraocular muscle tension during strabismus surgery. Some opioids had the same augmenting magnitude as the blocking effect of IV anticholinergic medication. Meperidine, which has some anticholinergic characteristics, neither blocked nor augmented the OCR. PMID- 15530139 TI - Myasthenia gravis in childhood. AB - INTRODUCTION: Pathologic alterations in neuromuscular transmission, known as Myasthenia Gravis (MG), comprise combinations (forms) of pre- and post-synaptic changes whose age at onset and clinical manifestations are dissimilar. Some forms have autoimmune features, while others have a defect due to functional or structural plaque alternations. The purpose of this work is to describe the clinical forms of MG seen in childhood, to facilitate proper diagnosis and treatment. MATERIAL AND METHODS: Retrospective data from 87 patients studied at the Ricardo Gutierrez Children's Hospital from 1990 to 2002 are presented. They were evaluated by clinical, pharmacological, electrophysiological, laboratory and radiological exams. RESULTS: Three distinct groups of patients were found: Group 1, Transient Neonatal Myasthenia Gravis (TNMG) represented 6.8% 9 (n=6) of the total number of patients. None of these six patients had ocular involvement. Group 2, Congenital Myasthenia (CMG) represented 17.2% of cases 9 (n=15). These patients had an onset of symptoms from 1 to 12 months of age. Eight cases were sporadic and the remainder familial. The most frequent symptom in this group was bilateral blepharoptosis in 13 patients. Group 3, Juvenile Myasthenia (JMG) comprised 76% of the patients (n=66). The onset of symptoms in this group ranged from 13 months to 14 years (mean 3.6 years). Thirty-nine of these children presented purely with ocular symptoms. CONCLUSIONS: Children with myasthenia gravis may present to the ophthalmologist first, with symptoms such as unilateral or bilateral blepharoptosis, diplopia, strabismus or ophthalmoplegia. Therefore, it is essential that the ophthalmologist be aware of the varied features of this disorder in children in order to diagnosis it correctly and make appropriate treatment referrals. PMID- 15530140 TI - Dynamic aniseikonia measurement: prismatic effect appears on the hess chart. AB - PURPOSE: To determine if aniseikonia appears on the Hess [Screen] Chart and if it can be measured thereby; and to compare such measurement to the standard Awaya New Aniseikonia Test in both artificial and pathologic aniseikonia. SUBJECTS AND METHODS: Twenty normal subjects with 20% artificial induced aniseikonia (Group 1) and 21 patients with pathologic aniseikonia (Group 2) were examined with the New Aniseikonia Test (NAT) and the Hess Chart. Group 2 was divided into three subgroups: Group 2-1, 13 patients with aphakia, whose operated eyes were more hypermetropic that their better eyes; Group 2-2, 3 patients with myopic anisometropia more than 5 diopters; and Group 2-3, 5 patients who had received macular translocation surgery (MTS). RESULTS: In normals, measurement of the 20% artificial aniseikonia ranged from 10% to 21% (mean 15%) using the NAT, and from 16% to 39% (mean 27.1% vertically and 24.6% horizontally) using the Hess Chart. In the 21 patients with pathologic aniseikonia, the NAT measurements ranged from 0% to 24% (upper limit of the NAT) and the Hess Chart measurements ranged from 0% to 65%. A large amount of the aniseikonia that appeared on the Hess Chart disappeared or was significantly reduced by changing the spectacle correction to a contact lens or intraocular lens. CONCLUSION: The aniseikonia that appeared on the Hess Chart was dynamic aniseikonia, due to rotational magnification inherent and unavoidable in the execution of the aniseikonia measurement on the Hess Chart. Aniseikonia, not due to refractive errors brought on by retinal surgery such as MTS, is difficult to correct with iseikonic lenses or other optical means. PMID- 15530141 TI - Inferior oblique overaction/contracture following retrobulbar anesthesia for cataract extraction with a positive Bielschowsky Head Tilt test to the contralateral shoulder. A report of one case. AB - BACKGROUND AND PURPOSE: This is a report of a patient with diplopia, hypertropia, extorsion and a positive Bielschowsky Head Tilt Test following retrobulbar anesthesia, due to an inferior oblique muscle overaction-contracture. CASE REPORT: Oculomotor and sensorimotor examinations and ocular motility recordings and Bielschowsky Head Tilt Test demonstrated overaction characteristics overpowering the contracture characteristics of the involved inferior oblique muscle. RESULTS: Weakening of the overacting inferior oblique eliminated the vertical and torsional deviations and the forced head tilt difference upon tilting the head to either shoulder. It also normalized ocular motility and resulted in a symptom-free patient. CONCLUSION: Oblique muscle dysfunction with vertical and torsional deviations and a positive Bielschowsky Head Tilt Test can result from retrobulbar anesthesia. PMID- 15530143 TI - LDL heterogeneity revisited: lesson for the metabolic syndrome from a new interpopulation study? PMID- 15530142 TI - Regulation of skeletal ryanodine receptors by dihydropyridine receptor II-III loop C-region peptides: relief of Mg2+ inhibition. AB - The aim of the present study was to explore interactions between surface-membrane DHPR (dihydropyridine receptor) Ca2+ channels and RyR (ryanodine receptor) Ca2+ channels in skeletal-muscle sarcoplasmic reticulum. The C region (725Phe-Pro742) of the linker between the 2nd and 3rd repeats (II-III loop) of the a1 subunit of skeletal DHPRs is essential for skeletal excitation-contraction coupling, which requires a physical interaction between the DHPR and RyR and is independent of external Ca2+. Little is known about the regulatory processes that might take place when the two Ca2+ channels interact. Indeed, interactions between C fragments of the DHPR (C peptides) and RyR have different reported effects on Ca2+ release from the sarcoplasmic reticulum and on RyR channels in lipid bilayers. To gain insight into functional interactions between the proteins and to explore different reported effects, we examined the actions of C peptides on RyR1 channels in lipid bilayers with three key RyR regulators, Ca2+, Mg2+ and ATP. We identified four discrete actions: two novel, low-affinity (>10 microM), rapidly reversible effects (fast inhibition and decreased sensitivity to Mg2+ inhibition) and two slowly reversible effects (high-affinity activation and a slow-onset, low-affinity inhibition). Fast inhibition and high-affinity activation were decreased by ATP. Therefore peptide activation in the presence of ATP and Mg2+, used with Ca2+ release assays, depends on a mechanism different from that seen when Ca2+ is the sole agonist. The relief of Mg2+ inhibition was particularly important since RyR activation during excitation-contraction coupling depends on a similar decrease in Mg2+ inhibition. PMID- 15530144 TI - Down-regulation of sarcolipin mRNA expression in chronic atrial fibrillation. AB - BACKGROUND: Abnormal intracellular Ca2+ homeostasis is an important modulator of chronic atrial fibrillation. Sarcolipin, a homologue of phospholamban, is specifically expressed in the atria, and may play an important role in modulating intracellular Ca2+ homeostasis in the atria. The aim of this study was to investigate the expression of sarcolipin mRNA in the atrial myocardium of patients with chronic atrial fibrillation. METHODS: We analyzed the expression of sarcolipin, phospholamban, cardiac calsequestrin and sodium calcium exchanger mRNAs in the right atrial myocardium from nine patients with mitral valvular disease with atrial fibrillation (MVD/AF), nine patients with MVD who had normal sinus rhythm (MVD/NSR), and 10 control patients with normal sinus rhythm who received open heart surgery (controls). The expression of mRNA was measured using the ABI PRISM 7700 Sequence Detection System (Applied Biosystems, Foster City, CA). RESULTS: Relative expression levels of sarcolipin mRNA were significantly lower in MVD/AF (0.60 +/- 0.11) than in either MVD/NSR (1.28 +/- 0.17, P < 0.01) or controls (1.10 +/- 0.10, P < 0.05). The expression levels of sarcolipin mRNA were significantly lower in the group with high values for right atrial pressure. The expression levels of phospholamban, cardiac calsequestrin and sodium calcium exchanger mRNAs were comparable among all three groups. CONCLUSIONS: Chronic electrical and mechanical overload decreased the expression of sarcolipin mRNA in the right atrial myocardium in patients with chronic atrial fibrillation. Down regulation of sarcolipin mRNA may be part of atrial fibrillation-induced atrial remodelling. PMID- 15530145 TI - Oxygen consumption is increased relative to work rate in patients with McArdle's disease. AB - BACKGROUND: Patients with McArdle's disease suffer exercise incapacity as a result of myophosphorylase deficiency, and for a given work rate have excessive circulatory and ventilatory responses. We hypothesized that the rate of increase of oxygen consumption with work rate (DeltaVO2-DeltaWR slope) would also be elevated in such patients as a result of these excessive responses. PATIENTS AND METHODS: Five patients with McArdle's disease and five matched controls carried out a maximal incremental cardiopulmonary exercise test. Controls then carried out a second test matched to the maximal test of a paired patient. Venous blood was sampled at rest, peak exercise and recovery. RESULTS: During the matched test, the DeltaVO2-DeltaWR slope was higher in the patients than in the controls [19.9 (15.0-24.6) vs. 11.7 (9.2-13.5) mL min(-1) W(-1); mean (range); P = 0.022], and the peak-achieved VO2 was also greater in the patient group [1201 (890-1575) vs. 918 (599-1248) mL min(-1); P = 0.003]. A similar pattern was observed for heart rate [173 (165-182) vs. 108 (105-134) b.p.m.; P = 0.001] and plasma norepinephrine levels [12.6 (9.2-19.9) vs. 2.9 (2.2-4.9) nmol l(-1); P = 0.003]. CONCLUSION: There is an increased rate of rise in VO2 relative to work rate during exercise in patients with McArdle's disease. There is also a greater rise in catecholamines, which may be the result of a physiological response to substrate starvation, and is likely to contribute to the increase in VO2. PMID- 15530146 TI - Ethnic differences in the formation of small LDL particles in Asians: a comparison of Koreans, Japanese and Mongolians. AB - BACKGROUND: The presence of small, dense LDL particles has been recognized as an independent risk factor for coronary heart disease (CHD) but is not directly representative of CHD mortality rate beyond any given population. We investigated whether such inconsistency between three Asian ethnic groups might have arisen from anthropometric and metabolic factors. DESIGN: We conducted a cross-sectional survey among adult Koreans (412), Japanese (453) and Mongolians (253). RESULTS: The prevalence of small LDL particles was 36% in the Koreans, 21% in the Japanese and 7% in the Mongolians. Multiple logistic regression analysis revealed plasma triglyceride (TG) levels to be the strongest determinant of small LDL particle size in all three groups, with sex, HDL-cholesterol and non-HDL-C being other ethnic-specific significant determinants. Body mass index (BMI), FFA and insulin resistance were not significant factors in the regression analysis. Of the subjects with low TG levels (< 133 mg dL(-1)), 25% of the Koreans and 10% of the Japanese, but no Mongolians, had small LDL particles. CONCLUSIONS: Results of the present study suggest that traditionally, high-carbohydrate diets in Korea and Japan possibly contribute to higher TG-levels compared with BMI-matched Mongolians, and to the formation of small LDL particles, even in instances of low TG levels. PMID- 15530147 TI - Chemical-physical properties of lipoproteins in anorexia nervosa. AB - BACKGROUND: Anorexia nervosa (AN), a psychosomatic disorder, has serious negative effects on multiple organs and systems of the human body. Anorexia nervosa usually runs a chronic course and is associated with significant morbidity and mortality. In order to elucidate the role played by lipids in AN, in the present study we compared the plasma lipid profile and the chemical-physical properties of lipoproteins obtained from subjects affected by AN. MATERIALS AND METHODS: The study was performed on lipoproteins of AN subjects and of age-matched healthy subjects used as controls. We tested the susceptibility to oxidative stress in vitro, the fatty acid content, the fluidity using 2-dimethylamino-(6-lauroyl) naphthalene (Laurdan) and 1,6-difenil-1,3,5-esatriene (DPH) probes. RESULTS: Present results indicate that AN patients present a deep alteration of the composition and of chemical-physical properties in circulating lipoproteins, even in the absence of significant modifications to clinical metabolic parameters. A significantly decreased body mass index (BMI) was found in AN patients in comparison with controls. Anorexia nervosa patients showed a significant modification of phospholipids to protein ratio and a significantly increased percentage of unsaturated fatty acids compared with control subjects as well as a decreased fluidity, a significantly increased percentage of liquid-crystalline phase in VLDL, and a significantly reduced susceptibility to oxidative stress, more pronounced in LDL. CONCLUSIONS: These results confirm the hypothesis that anorexia is accompanied by changes of lipid metabolism in the central nervous system (CNS). PMID- 15530148 TI - Autoantibodies to REG, a beta-cell regeneration factor, in diabetic patients. AB - BACKGROUND: Regenerating gene (Reg) product, Reg, acts as an autocrine/paracrine growth factor for beta-cell regeneration. The presence of autoimmunity against REG may affect the operative of the regenerative mechanisms in beta cells of Type 1 and Type 2 diabetes patients. We screened sera from Type 1 and Type 2 diabetes subjects for anti-REG autoantibodies, searched for correlations in the general characteristics of the subjects with the presence of anti-REG autoimmunity, and tested the attenuation of REG-induced beta-cell proliferation by the autoanitibodies. MATERIAL AND METHODS: We examined the occurrence of anti-REG autoantibodies in patients' sera (265 Type 1, 368 Type 2 diabetes patients, and 75 unrelated control subjects) by Western blot analysis, and evaluated inhibitory effects of the sera on REG-stimulated beta-cell proliferation by a 5'-Bromo-2' deoxyuridine (BrdU) incorporation assay in vitro. RESULTS: Anti-REG autoantibodies were found in 24.9% of Type 1, 14.9% of Type 2 and 2.7% of control subjects (P = 0.0004). There were significant differences between the autoantibody positive and negative groups in the duration of disease in the Type 1 subjects (P = 0.0035), and the age of onset in the Type 2 subjects (P = 0.0274). The patient sera containing anti-REG autoantibodies significantly attenuated the BrdU incorporation by REG (35.6 +/- 4.06% of the control), whereas the nondiabetic sera without anti-REG autoantibodies scarcely reduced the incorporation (88.8 +/- 5.10%). CONCLUSION: Anti-REG autoantibodies, which retard beta-cell proliferation in vitro, are found in some diabetic patients. Thus, autoimmunity to REG may be associated with the development/acceleration of diabetes in at least some patients. PMID- 15530149 TI - Treatment with granulocyte colony-stimulating factor increases interleukin-1 receptor antagonist levels during engraftment following allogeneic stem-cell transplantation. AB - BACKGROUND: The effect of treatment with granulocyte colony-stimulating factor (G CSF) on interleukin-1 receptor antagonist (IL-1ra) plasma concentrations as well as the role of IL-1ra on leucocyte recovery and parameters of infection within the first 30 days after haematopietic stem-cell transplantation (HSCT) are not well known. MATERIAL AND METHODS: Twenty-seven patients undergoing myeloablative therapy followed by allogeneic SCT for various haematological disorders were either treated with (n = 18) or without (n = 9) G-CSF. IL-1ra plasma levels were serially determined by ELISA starting at day - 1 and continued until patients were engrafted. RESULTS: Patients receiving G-CSF had significantly shorter neutropenic periods and significantly lower mean C-reactive protein serum levels during the first 3 weeks succeeding bone marrow transplantation (BMT). Importantly, starting at day + 11 and paralleling the rise of peripheral blood leucocytes, increasing IL-1ra plasma concentrations were observed in both treatment groups. However, the magnitude of the IL-1ra surge was far greater in the G-CSF treatment group. Peak IL-1ra plasma level observed on day + 19 was 882.3 +/- 879.2 pg mL(-1) (mean +/- SD) in patients receiving G-CSF compared with 285.8 +/- 175.2 pg mL(-1) (mean +/- SD) in patients not receiving G-CSF (P = 0.0130). Furthermore, a direct correlation of IL-1ra with peripheral blood leucocytes was verified by the Spearman rank test (P = 0.0025). CONCLUSION: Granulocyte colony-stimulating factor-mediated acceleration of neutrophil recovery following myeloablative therapy correlated with increased IL-1ra plasma concentrations. Our data suggest that IL-1ra constitutes an intrinsic component of the anti-inflammatory and neutrophil differentiating efficacy of G-CSF and, thus, IL-1ra may be required for the in vivo activity of G-CSF. PMID- 15530150 TI - Weight reduction, but not a moderate intake of fish oil, lowers concentrations of inflammatory markers and PAI-1 antigen in obese men during the fasting and postprandial state. AB - BACKGROUND: In obese subjects, chronic low-grade inflammation contributes to an increased risk of metabolic abnormalities, which are reversed by weight loss. Sustained weight loss, however, is difficult to achieve and more insight into dietary approaches on anti-inflammatory responses in obese subjects is needed. In this respect, fish oil deserves attention. MATERIAL AND METHODS: Eleven obese men (BMI: 30-35 kg m(-2)) received daily fish oil (1.35 g n-3 fatty acids) or placebo capsules in random order for 6 weeks. Eight subjects continued with a weight reduction study that lasted 8 weeks. Mean weight loss was 9.4 kg. At the end of each experimental period a postprandial study was performed. RESULTS: Relative to fasting concentrations, interleukin-6 (IL-6) levels increased by 75% 2 h and by 118% 4 h after the meal (P < 0.001), when subjects consumed the control capsules. In contrast, C-reactive protein (C-RP) concentrations decreased slightly by 0.7% and 6.6% (P = 0.046), and those of plasminogen activator inhibitor-1 (PAI-1) antigen by, respectively, 26% and 53% (P < 0.001). Tumour necrosis factor-alpha (TNF-alpha; P = 0.330) and soluble TNF-receptor concentrations (sTNF-R55 and sTNF R75; P = 0.451 and P = 0.108, respectively) did not change. Changes relative to fasting concentrations were not significantly affected by either fish oil or weight reduction. Absolute IL-6, C-RP, sTNF-R55, sTNF-R75, and PAI-1 antigen concentrations, however, were consistently lower after weight reduction, but not after fish oil consumption. CONCLUSION: For slightly obese subjects a moderate intake of fish oil does not have the same favourable effects on markers for a low grade inflammatory state as weight reduction. PMID- 15530151 TI - Retinol binding protein isolated from acute renal failure patients inhibits polymorphonuclear leucocyte functions. AB - BACKGROUND: Protein factors accumulating in sera of patients with end-stage renal disease (ESRD) that interfere with the nonspecific immune response by inhibiting essential functions of polymorphonuclear leucocytes (PMNLs) have previously been described. No such factor has been isolated from acute renal failure (ARF) patients to date. MATERIALS AND METHODS: Using a three-step chromatographic procedure involving ion exchange, size exclusion and hydrophobic interaction chromatography we purified the apo- and holo-form of retinol binding protein (RBP) from high-flux dialyser (polyacrylonitrile; AN69) ultrafiltrates of patients with ARF. Their effect on the chemotaxis of PMNLs isolated from healthy donors was determined by the under-agarose method. Whole-blood assays applying flow cytometry were used to assess phagocytosis and the oxidative metabolism of PMNLs. Apoptosis was assessed by determining the DNA content using propidium iodide. RESULTS: Isolated apo- and holo-forms of RBP were truncated on their C terminus as determined by mass spectrometry. All isolates significantly inhibited the chemotactic movement of PMNLs obtained from healthy donors and the PMNL oxidative metabolism stimulated by E. coli. These effects were concentration dependent. Retinol binding protein had no influence on the PMNL oxidative metabolism stimulated by PMA and on PMNL phagocytosis. Commercially available RBP isolated from urine influenced PMNL functions in the same way. Inhibition of p38 mitogen-activated protein kinase (MAPK) by SB203580 significantly attenuated the phagocytosis-induced respiratory burst and RBP did not lead to a further decrease. Polymorphonuclear leucocyte apoptosis was significantly inhibited by RBP. CONCLUSIONS: The apo- and holo-forms of RBP isolated from the ultrafiltrate of ARF patients inhibit PMNL chemotaxis, oxidative metabolism and apoptosis. Therefore, RBP may be considered a uraemic toxin contributing to a disturbed immune defence. PMID- 15530152 TI - Oral ferrous sulphate leads to a marked increase in pro-oxidant nontransferrin bound iron. PMID- 15530154 TI - Promoting mental health nursing research in low and middle income countries. PMID- 15530155 TI - Nurse migration is only a symptom, not the disease. Interview by Jan Harrington. PMID- 15530159 TI - Human resources - we need new ideas. PMID- 15530160 TI - Tackling drug and alcohol misuse in Brazil: priorities and challenges for nurses. AB - AIMS: To provide an overview of the extent of drug and alcohol misuse in Brazil and the policies and approaches in tackling substance misuse. An examination of the challenges facing the nursing profession in working with substance misusers is presented. BACKGROUND: Alcohol, cocaine, and cannabis are the most commonly misused psychoactive substances in Brazil. One of the biggest public health problems is the interface between the misuse of psychoactive substances and HIV prevalence and other sexually transmitted diseases. Findings from a recent study suggest that undergraduate nurses in Brazil are not adequately prepared in the care and management of substance misuse problems. The nursing profession in Brazil faces numerous challenges in the development of professional competence of nurses in this field. CONCLUSION: A strategy proposed is the creation of regional centres in Brazil to study the integration of substance use and misuse in the nursing undergraduate curriculum and the giving of specific support in teaching and research to nurse teachers. Nurses have a key role to play in the early recognition, assessment, prevention, and treatment of substance misuse. PMID- 15530161 TI - Monitoring adverse drug reactions: scales, profiles, and checklists. AB - BACKGROUND: Globally, adverse drug reactions (ADRs) make a substantial contribution to ill health. Introducing a systematic approach to patient surveillance could mitigate these problems. Formalized medication monitoring schedules have been proposed as one strategy to diagnose and action side-effects and the problems emanating from adverse drug reactions. To date, most developments have been linked to antipsychotic medications. Several scales, checklists, and side-effect profiles are available, including the West Wales ADR (adverse drug reaction) profile. However, relatively little work has been undertaken on the clinical validity, reliability, and sensitivity of these instruments. AIM: This paper describes the development of the monitoring schedule approach to medication management. It also reviews and compares the instruments available for monitoring the adverse drug reactions of antipsychotic medications. The UKU (Udvalg for Kliniske Undersogelser) scale and the West Wales ADR profile assess a broader range of physiological parameters and potential problems than other instruments. However, to be adopted in practice, such instruments must achieve a balance between clinical gain and practical cost, including the time spent in administration. CONCLUSION: Further work is needed to explore the translation of formalized ADR surveillance programmes into clinical gains and improved outcomes for clients. PMID- 15530162 TI - Self-harm in young people: a perspective for mental health nursing care. AB - AIMS: This paper addresses the assessment and management of self-harm in young people by a nurse consultant-led self-harm service in the UK. The purpose of the paper is to present some of the theoretical perspectives of the service and how these have influenced current practice with this group of young people. BACKGROUND: Self-harm in young people continues to be a serious concern for health services globally. Rates of self-harm in teenagers in the UK are among the highest in Europe. The current literature offers information on the nature of self-harm and some evidence of specific interventions mainly from a 'psychiatric perspective'. This paper examines the development of a locally based nurse-led self-harm team involved in the care of this group of young people. A sociological view of children and young people is applied in examining the way society construes self-harm and the formation of the behaviour as a deviant act. Self harm is considered in the context of transition through adolescence and the growth of self-identity. CONCLUSION: The locally based child and adolescent mental health service has drawn together the evolving nurse consultant role, mental health nursing and medical sociology. This integration of people and theory will enhance the services understanding of self-harm in young people. At an international level, there is a need to consider the sociological dimensions of self-harm in young people as a way forward for mental health nurses who are involved in working with this group of people. PMID- 15530163 TI - Characteristics of and interventions for fever in Japan. AB - PURPOSE: As part of a larger multinational validation study of the International Classification for Nursing Practice (ICNP) alpha version, a survey was conducted in Japan to determine characteristics of 'fever' and interventions to treat febrile patients. SAMPLE: Three hundred and fifty-six acute and critical care Japanese nurses participated in this study. METHOD: The major and minor characteristics of 'fever' perceived by Japanese nurses and interventions used by the nurses in managing febrile patients were identified using the Diagnostic Content Validity (DCV) model. RESULTS: Two characteristics, 'increased body temperature' and 'chills' were selected as major characteristics from the standardized list of the ICNP alpha version validation study. Nine characteristics among the standardized list of characteristics were rated as minor characteristics, and six of the ICNP characteristics were rejected. 'Shivering' and 'infectious lab data' were added with a level of representativeness similar to a major characteristic by nine of the nurses. A variety of interventions to treat fever were reported. The most frequently reported intervention was cooling, followed by warming and medication. Nine dimensions were derived from all reported interventions. DISCUSSION: As well as perceived characteristics of fever, these interventions may have aspects unique to Japanese nursing practice and to the acute and critical care settings. These results can be compared to those of other populations in future studies. PMID- 15530164 TI - Nursing diagnosis with the ICNP in the teaching context. AB - BACKGROUND: The premise for the research study was the authors' conviction that the use of nursing diagnostic taxonomies allows the nurse to formulate a thorough and valid diagnosis. AIM: To verify the International Classification for Nursing Practice (ICNP) in the context of a Polish academic nursing programme. RESEARCH QUESTIONS: Do nursing diagnoses made traditionally and those made with the ICNP reflect the patient's/client's condition? Is the range and type of the two kinds of diagnoses similar or different? Is the level of detail in both types of diagnoses compatible? METHODS: Quasi-experiment in which the experimental group diagnosed patients by means of the ICNP and the control group formulated nursing diagnoses by means of an intellectual decision-making process. STUDY GROUPS: 44 students in a MNurs. Programme, Faculty of Nursing and Health Sciences, Medical University in Lublin, Poland. CONCLUSIONS: The range and type of nursing diagnoses do not depend on the method used in their formulation. The ICNP diagnoses are compatible with the traditionally formulated ones. The unbalanced size of the experimental and the study groups negatively influenced the validity of conclusions. PMID- 15530165 TI - Effect of 50% ethanolic extract of Syzygium aromaticum (L.) Merr. & Perry. (clove) on sexual behaviour of normal male rats. AB - BACKGROUND: The flower bud of Syzygium aromaticum (L.) Merr. & Perry. (clove) has been used in Unani medicine since ancient times for the treatment of male sexual disorders. The present study is aimed to investigate the effect of 50% ethanolic extract of clove on general mating behaviour, libido, potency along with its likely gastric ulceration and adverse effects on sexually normal male albino rats. METHODS: The suspension of the extract was administered orally at the dose of 100, 250, and 500 mg / kg, to different groups of male rats (n = 6) once a day for seven days. The female albino rats involved in mating were made receptive by hormonal treatment. The general mating behaviour, libido and potency were determined and compared with the standard reference drug sildenafil citrate. The probable gastric ulceration and adverse effects of the extract were also evaluated. RESULTS: Oral administration of the extract significantly increased the Mounting Frequency, Intromission Frequency; Intromission Latency, Erections; Quick Flips, Long Flips as well as aggregate of penile reflexes and caused significant reduction in the Mounting Latency and Post Ejaculatory Interval. The most appreciable effect of the extract was observed at the dose of 500 mg/kg. The test drug was also found to be devoid of any conspicuous gastric ulceration and adverse effects. CONCLUSION: The results indicated that the 50% ethanolic extract of clove produced a significant and sustained increase in the sexual activity of normal male rats, without any conspicuous gastric ulceration and adverse effects. Thus, the resultant aphrodisiac effectivity of the extract lends support to the claims for its traditional usage in sexual disorders. PMID- 15530166 TI - The impact of two multiple-choice question formats on the problem-solving strategies used by novices and experts. AB - BACKGROUND: Pencil-and-paper examination formats, and specifically the standard, five-option multiple-choice question, have often been questioned as a means for assessing higher-order clinical reasoning or problem solving. This study firstly investigated whether two paper formats with differing number of alternatives (standard five-option and extended-matching questions) can test problem-solving abilities. Secondly, the impact of the alternatives number on psychometrics and problem-solving strategies was examined. METHODS: Think-aloud protocols were collected to determine the problem-solving strategy used by experts and non experts in answering Gastroenterology questions, across the two pencil-and-paper formats. RESULTS: The two formats demonstrated equal ability in testing problem solving abilities, while the number of alternatives did not significantly impact psychometrics or problem-solving strategies utilized. CONCLUSIONS: These results support the notion that well-constructed multiple-choice questions can in fact test higher order clinical reasoning. Furthermore, it can be concluded that in testing clinical reasoning, the question stem, or content, remains more important than the number of alternatives. PMID- 15530167 TI - Recombinant activated factor VII (rFVIIa) as salvage treatment for intractable hemorrhage. AB - BACKGROUND: Recently, there has been an increased use of recombinant activated factor VII (rFVIIa) to promote hemostasis in various hemorrhagic conditions. The objective of this study was to determine the outcome of patients treated with rFVIIa who had intractable bleeding associated with cardiac surgery (CSP) or as a result of other causes (OBP). METHODS: The medical records of 40 consecutive patients treated with rFVIIa were retrospectively reviewed for blood product use before and after treatment. In all patients, rFVIIa was given only after all other measures to stop bleeding had failed. The number of transfused units of red cells (R), platelets (P), fresh frozen plasma (F), and cryoprecipitate (C) were determined both before and after administration of rFVIIa, and the results compared. Mortality at 4 hours and 30 days was assessed. Patients dying within 4 hours of rFVIIa administration were not evaluable for response. Patient characteristics were also assessed as risk factors for mortality. RESULTS: Twelve of 24 CSP survived for more than 4 hours. These 12 patients required an average of 17 units (U) of R, 18 U of P, 18 U of F and 15 U of C pre-treatment compared to an average of 6 U, 10 U, 9 U and 4 U of R, P, F and C respectively, post treatment. These differences were statistically significant. For the OBP, 11 of 16 survived more than four hours. These 11 patients required an average of 10 U of R, 11 U of P, 14 U of F and 10 U of C pretreatment compared to an average of 1 U, 2 U, 2 U and 0 U of R, P, F, and C respectively, post-treatment. With the exception of C, there was a statistically significant decrease in blood product use following treatment with rFVIIa. Of the survivors in each group, 6 of 12 CSP and 2 of 11 OBP died between 3 and 30 days post-treatment from causes other than bleeding. Mortality at 30 days for CSP and OBP survivors was 50% and 18% respectively, whereas overall 30 day mortality was 75% for CSP and 44% for OBP. CONCLUSIONS: rFVIIa is effective in decreasing blood product use and promoting hemostasis in patients with intractable bleeding associated with cardiac surgery and a variety of other causes. PMID- 15530169 TI - Older age does not influence CD4 cell recovery in HIV-1 infected patients receiving highly active antiretroviral therapy. AB - BACKGROUND: Diagnosis of HIV infection is recently occurring with increasing frequency in middle-aged and in older individuals. As HAART became available, a minimal beneficial effect on immunological outcome in older in respect of younger subjects has been reported. In fact, both the intensity and the rapidity of the immunological response appeared to be reduced in elderly subjects. On the contrary, only few reports have indicated a similar immunological outcome both in older and younger HIV-positive subjects. Interestingly, older age did not seem to significantly affect the long-term virological outcome of HAART treated subjects. METHODS: To characterise epidemiological and clinical features of older HIV+ subjects, a prospective case-control study was performed: 120 subjects >/= 50 and 476 between 20 and 35 years were initially compared. Subsequently, to better define the impact of HAART on their viro-immunological response, 81 older were compared with 162 younger subjects. RESULTS: At baseline cases presented significantly lower TCD4+ cell number and were more frequently affected by comorbid conditions. Under HAART a statistically significant increase in TCD4+ cell number was observed in cases and controls. At multivariate analysis, there was no statistically significant difference between cases and controls regarding viro-immunological response. CONCLUSIONS: Although older subjects present a more severe HIV infection, they can achieve, under HAART, the same viro-immunological success as the younger individuals. PMID- 15530170 TI - A survey of relationship between anxiety, depression and duration of infertility. AB - BACKGROUND: A cross sectional study was designed to survey the relationship between anxiety/depression and duration/cause of infertility, in Vali-e-Asr Reproductive Health Research Center, Tehran, Iran. METHODS: After obtaining their consents, 370 female patients with different infertility causes participated in, and data gathered by Beck Depression Inventory(BDI) and Cattle questionnaires for surveying anxiety and depression due to the duration of infertility. This was studied in relation to patients' age, educational level, socio-economic status and job (patients and their husbands). RESULTS: Age range was 17-45 years and duration and cause of infertility was 1-20 years. This survey showed that 151 women (40.8%) had depression and 321 women (86.8%) had anxiety. Depression had a significant relation with cause of infertility, duration of infertility, educational level, and job of women. Anxiety had a significant relationship with duration of infertility and educational level, but not with cause of infertility, or job. Findings showed that anxiety and depression were most common after 4-6 years of infertility and especially severe depression could be found in those who had infertility for 7-9 years. CONCLUSIONS: Adequate attention to these patients psychologically and treating them properly, is of great importance for their mental health and will improve quality of their lives. PMID- 15530171 TI - Efficacy of repeated intrathecal triamcinolone acetonide application in progressive multiple sclerosis patients with spinal symptoms. AB - BACKGROUND: There are controversial results on the efficacy of the abandoned, intrathecal predominant methylprednisolone application in multiple sclerosis (MS) in contrast to the proven effectiveness in intractable postherpetic neuralgia. METHODS: We performed an analysis of the efficacy of the application of 40 mg of the sustained release steroid triamcinolone acetonide (TCA). We intrathecally injected in sterile saline dissolved TCA six times within three weeks on a regular basis every third day in 161 hospitalized primary and predominant secondary progressive MS patients with spinal symptoms. The MS patients did not experience an acute onset of exacerbation or recent distinct increased progression of symptoms. We simultaneously scored the MS patients with the EDSS and the Barthel index, estimated the walking distance and measured somatosensory evoked potentials. Additionally the MS patients received a standardized rehabilitation treatment. RESULTS: EDSS score and Barthel index improved, walking distance increased, latencies of somatosensory evoked potentials of the median and tibial nerves shortened in all MS patients with serial evaluation (p < 0.0001 for all variables). Side effects were rare, five patients stopped TCA application due to onset of a post lumbar puncture syndrome. CONCLUSIONS: Repeated intrathecal TCA application improves spinal symptoms, walking distance and SSEP latencies in progressive MS patients in this uncontrolled study. Future trials should evaluate the long-term benefit of this invasive treatment. PMID- 15530168 TI - Role of a critical visceral adipose tissue threshold (CVATT) in metabolic syndrome: implications for controlling dietary carbohydrates: a review. AB - There are likely many scenarios and pathways that can lead to metabolic syndrome. This paper reviews mechanisms by which the accumulation of visceral adipose tissue (VAT) may contribute to the metabolic syndrome, and explores the paradigm of a critical VAT threshold (CVATT). Exceeding the CVATT may result in a number of metabolic disturbances such as insulin resistance to glucose uptake by cells. Metabolic profiles of patients with visceral obesity may substantially improve after only modest weight loss. This could reflect a significant reduction in the amount of VAT relative to peripheral or subcutaneous fat depots, thereby maintaining VAT below the CVATT. The CVATT may be unique for each individual. This may help explain the phenomena of apparently lean individuals with metabolic syndrome, the so-called metabolically normal weight (MONW), as well as the obese with normal metabolic profiles, i.e., metabolically normal obese (MNO), and those who are "fit and fat." The concept of CVATT may have implications for prevention and treatment of metabolic syndrome, which may include controlling dietary carbohydrates. The identification of the CVATT is admittedly difficult and its anatomical boundaries are not well-defined. Thus, the CVATT will continue to be a work in progress. PMID- 15530172 TI - Barriers to providing osteoporosis care in skilled nursing facilities: perceptions of medical directors and directors of nursing. AB - OBJECTIVES: The objectives of this study were to identify the barriers to osteoporosis clinical practice guideline use perceived by Medical Directors (MED DIR) and Directors of Nursing (DON) in skilled nursing facilities; and to describe differences in the perceptions of MED DIRs and DONs. DESIGN: The authors conducted a cross-sectional national survey. PARTICIPANTS: This study consisted of a random national sample of MED DIRs (n=1300) and DONs (n=1300) belonging to the American Medical Directors Association or the National Association of Directors of Nursing Administration in Long-term Care. MEASUREMENTS: A 24-item survey using a five-point Likert scale was developed. The survey measured agreement to questions in four domains (provider factors, guideline characteristics, patient factors, environmental factors) and 10 content areas (problem acknowledgment, patient/family concern, patient/family compliance, testing availability, safety, reimbursement, regulatory oversight, staff knowledge/time/ability, belief in guidelines, and malpractice liability). Response distributions to each item were plotted and differences between MED DIRs and DONs were tested. RESULTS: Survey response rates were 40% for MED DIRs and 48% for DONs. Respondents strongly agreed that fractures are a problem in their facilities and that osteoporosis guidelines are useful and cost-beneficial (mean responses > or = 4.0). A large proportion of respondents (at least 40% of the sample) identified multiple patient comorbidities, reimbursement issues, length of stay, and regulatory oversight as barriers to providing osteoporosis care. Respondents did not believe that patient and family acceptance, testing availability, staff time, staff self-efficacy, or concerns about bisphosphonate safety were barriers to osteoporosis care. DONs were more likely than MED DIRs to believe that patients and families are concerned about fractures, whereas MED DIRs were more likely to endorse length of stay, staffing issues, and regulatory oversight as influencing treatment decisions. Years of practice and facility size, but not formal geriatrics training, significantly influenced responses. CONCLUSION: Perceived barriers to implementing osteoporosis guidelines differ between facilities and between MED DIRs and DONs. Identification of these barriers could facilitate quality improvement initiatives and improve the quality of osteoporosis care. PMID- 15530174 TI - Identification of low bone mass in a developmental center: finger bone mineral density measurement in 562 residents. AB - OBJECTIVES: The objective of this study was to measure finger bone mineral density (BMD) in residents of a developmental center. DESIGN: A cross-sectional descriptive study performed during the residents' annual physical examination. SETTING: This study was conducted at a long-term care facility for people with severe developmental disabilities (mental retardation, cerebral palsy, epilepsy, and autism). PARTICIPANTS: Study participants included 562 (67%) of the 833 residents aged 30 years or more on whom we could measure BMD. MEASUREMENTS: We measured BMD by peripheral dual-energy x-ray absorptiometry (DXA) on the residents' middle fingers by Accudexa. The instrument converted the BMD values into T-scores relative to the manufacturer's reference young healthy population. We retrieved the residents' clinical variables from a centralized database. RESULTS: T-scores for the 562 residents were: <-2.5 standard deviation (SD) (osteoporotic): 98 (17%), -2.5 SD>t <-1 SD (osteopenic): 156 (28%), and >-1 SD (normal): 308 (55%). Multivariate regression analysis revealed that BMD was significantly lower in residents (compared with their counterparts) with inability to ambulate, male gender, white race, quadriplegia, profound mental retardation, and older age. CONCLUSION: There is a need for heightened osteoporosis surveillance and preventive effort in this population in which almost half of the residents had previously undiagnosed subnormal BMD, including one sixth who had osteoporosis. PMID- 15530173 TI - Low bone mass and postfall fracture risk among elderly nursing home men. AB - OBJECTIVE: To study postfall fracture risk in relation to bone mineral density. SETTING: The study was conducted in men residing in nursing homes. PARTICIPANTS: A total of 212 men above the age of 65 years were recruited, and of these, 172 satisfied the inclusion criteria of the study. Among them, 82 were less than 85 years old, and 90 were at least 85 years old. INTERVENTION: Bone mineral density (BMD) of the calcaneus was measured, using the Lunar PIXI Bone Densitometer (version 1.43) and charts were reviewed for age, falls, and fractures. MEASUREMENTS: The values of BMD were expressed as T-scores, and the Lunar criteria equivalents of T-scores were used (T-scores more than -0.6 for normal BMD, -0.6 to -1.6 for osteopenia, and below -1.6 for osteoporosis). RESULTS: The median age of all participants was 84 years (range 65 to 102 years), and their mean T-score at the calcaneus was -1.51. In about 51% of them, the T-score was in the osteoporotic range. For subjects less than 85 years old, the mean T-score was -1.19, and for those 85 years and older, the mean T-score was -1.81; the mean bone density of those in the older old group was significantly lower than those in the younger old group if there was a history of a fracture. Analysis by age quartiles showed a progressive decline in BMD with advancing age. About 53% of the subjects who fell (37.2%) sustained a nontraumatic fracture. Among them, 67.6% had osteoporosis. CONCLUSION: At any given age after age 65, men who sustained a postfall fracture had a significantly lower BMD than those who did not sustain a postfall fracture. PMID- 15530175 TI - The use of calcaneal ultrasound evaluation of bone mineral density in cognitively impaired seniors. AB - OBJECTIVES: Low bone mineral density (BMD) is a major risk factor for osteoporotic fracture. The gold standard measurement of BMD, central dual-energy x-ray absorptiometry (DXA), requires transportation to a radiologic facility, patient agility and cooperation. For those with severe cognitive impairment, this is not realistic; hence, central BMD is seldom measured. The objectives were to evaluate the use of calcaneal BMD measurement (by qualitative ultrasound), the prevalence of osteoporosis diagnosis (based on calcaneal BMD), and treatment in individuals with severe cognitive impairment. DESIGN: This is a point prevalence descriptive study. Calcaneal BMD was measured using a Sahara sonometer (Hologic). Cognition (Mini Mental Status Examination), mobility (Berg balance scale, timed up-and-go), and chart audit were also undertaken. SETTING: This study was conducted at four dementia long-term care facilities (122 beds) in Edmonton, Alberta, Canada. PARTICIPANTS: Forty long-term care residents participated in this study with an average MMSE of 10 (range, 0-25). MEASUREMENTS: One hundred percent of study subjects cooperated with calcaneal ultrasound measurement. RESULTS: Based on calcaneal BMD measurement using recommended cutoffs, 92.5% were at high risk for osteoporosis or osteopenia and 5% were at moderate risk. One participant (2.5%) had a normal calcaneal BMD measurement. Calcium supplementation occurred in 32% of the high-risk group, 50% of the moderate-risk group, and 100% of the normal group. One (2.5%) participant was on hormone replacement therapy, and 12% were taking etidronate. CONCLUSIONS: Calcaneal ultrasound measurement could be a useful tool to assess BMD in cognitively impaired nursing home residents. PMID- 15530176 TI - The importance of parathyroid hormone and vitamin D status in the treatment of osteoporosis and renal insufficiency. AB - Suboptimal vitamin D status is common in sick, elderly patients and is associated with secondary hyperparathyroidism, increased bone turnover, and accelerated bone loss. Nutritional supplementation of elderly patients with vitamin D and calcium has been associated with a reduction in fractures and falls. Recently, the National Kidney Foundation (NKF) recommended screening parathyroid hormone (PTH) levels in those with glomerular filtration rates (GFR) less than 60 mL/min. The PTH target range increases as renal failure progresses because of skeletal resistance. When PTH levels are above the target range, measurement of serum 25 hydroxyvitamin D (25-OH-D) is recommended. If the 25-OH-D level is less than 30 ng/mL, supplementation with vitamin D is recommended. Practitioners should consider supplementing most residents with vitamin D and calcium or follow NKF guidelines. PMID- 15530177 TI - Current approaches to postfall assessment in nursing homes. AB - OBJECTIVE: Falls in nursing homes occur among a large percentage of residents. Their onset necessitates a postfall assessment (PFA) be performed by clinical staff to determine likely etiology. The absence of an empirically validated comprehensive postfall assessment tool has led to considerable variability in the types of PFAs performed. The purpose of this study was to examine the types of PFA tools available, their content, and to compare this with national recommendations for fall assessment in geriatric practice. SETTING: A convenience sample of 379 long-term care facilities, with a cumulative census of over 40,000 residents, in New Jersey were solicited to return to the Director of Long-Term Care Surveys at the NJ Department of Health and Senior Services a copy of any PFA tools used in practice. METHODS: A review of the types of assessment tools used in each of the responding facilities were tabulated and coded as belonging to one of five categories: (1) fall-risk assessment short form, (2) fall-risk assessment long form, (3) fall prevention protocols such as fall programs and postfall assessment forms, (4) incident reports, and (5) other. A subset of 20 facilities used a specific PFA. This content was further analyzed and compared with national and professional recommendations for PFA that included five domains: (1) history of the fall, (2) environmental issues, (3) physical examination, (4) functional assessment, and (5) laboratory and other diagnostics. RESULTS: Of 379 facilities solicited, 149 responded (40%) to reveal a wide array of tools used for the purpose of PFA. These included: risk assessment tools, fall prevention programs, policies and procedures for fall management, and incident reports. Overall, most facilities used fall-risk assessment tools in place of PFA (63.7%; n=95). Many of the nationally recommended guidelines for PFA were not included in the tools included in this sample, with the exception of environmental questions that were evident in all PFAs surveyed (100%). Other fall circumstances related to time, mobility, and footwear were included less often (70% n=14) as were use of diuretics (55%;=11), mental status and ambulation ability (45%; n=9) of the falling older adult. CONCLUSION: Despite recommendations in the geriatric literature, comprehensive postfall assessment tools were unavailable for use by nursing home staff. When a PFA was performed, there was no consistency among facilities sampled. Data collected was minimal and unlikely to reveal the full range of possible underlying etiologies. Improved, validated PFA tools are needed to aid clinical staff in evaluating older adults who have fallen. PMID- 15530178 TI - Mild anemia and the risk of falls in older adults from nursing homes and the community. AB - OBJECTIVES: The objectives of this study were to determine if a relationship exists between a history of falls and anemia in older adults and to compare the findings by place of residence. DESIGN: The authors conducted a retrospective and observational study. PARTICIPANTS: One hundred forty-five adults (60-97 years of age) from nursing homes and the community hospitalized for hip fracture over a 2 year period were included in this study. MEASUREMENTS: Laboratory values (hemoglobin [Hb], hematocrit [Hct]), medical history, and demographics were measured. RESULTS: Falls occurred similarly in both nursing home patients and community patients (19% vs. 17%, P=0.785). Nursing home and community patients also had similar Hb (P=0.0899), Hct (P=0.1929), and rates of anemia (P=0.187). Nursing home residents were older (P=0.0188) and had lower serum albumin levels (P=0.0007) than community patients. When the two groups were combined, falls were more common in anemic individuals (30% vs. 13%; P=0.028). Furthermore, those with a history of falls were older (P=0.0447), had lower Hb (P=0.0257) and Hct levels (P=0.0310). After controlling for age, gender, place of residence, and arthritis in a logistic regression model, anemia predicted a threefold increased risk of falls (P=0.041), and a 45% decreased risk of falls was predicted for every 1.0 g/dL increase in Hb (P=0.005). Falls risk increased 7% per year of age (P=0.040), whereas musculoskeletal disease increased the falls risk 3.2-fold, both increases being independent of Hb levels or anemia. Finally, falls were not associated with gender or other comorbidity, nor did these variables alter the falls risk attributed to low Hb or anemia. CONCLUSIONS: These findings suggest a new and potentially important link between anemia and the risk of falls in patients sustaining hip fractures from both nursing homes and the community. Further studies will help determine if this risk is modified or eliminated with treatment of anemia and if the relationship applies to larger samples of older adults in different settings. PMID- 15530179 TI - Falls in the nursing home: are they preventable? AB - INTRODUCTION: Falls are prevalent in elderly patients residing in nursing homes, with approximately 1.5 falls occurring per nursing home bed-years. Although most are benign and injury-free, 10% to 25% result in hospital admission and/or fractures. Primary care providers for nursing home residents must therefore aim to reduce both the fall rate as well as the rate of fall-related morbidity in the long-term care setting. Interventions have been demonstrated to be successful in reducing falls in community-dwelling elderly patients. However, less evidence supports the efficacy of fall prevention in nursing home residents. METHODS: The authors conducted a Medline search using the key words Falls and Nursing Homes. RESULTS: Several studies examined the efficacy of multifaceted intervention programs on reducing falls in nursing homes with varied results. Components of these intervention programs include: environmental assessment, assistive device evaluation and modification, medication changes, gait assessment and training, staff education, exercise programs, hip protector use, and blood pressure evaluation. Current literature supports the use of environmental assessment and intervention in reducing falls in nursing homes, and demonstrates an association between certain medications and falls. However, there are no studies that examine the effect of medication adjustments on fall rates. Also, the literature does not strongly suggest that exercise programs are effective in fall reduction. Although not effective in reducing fall rates, the use of hip protectors appears to result in less fall-related morbidity. CONCLUSION: More studies must be done to clarify the effects of high-risk medication reduction, the optimal nature and intensity of exercise programs, and patient targeting criteria to maximize the effectiveness of nursing home fall prevention programs. Based on the current literature, an effective multifaceted fall prevention program for nursing home residents should include risk factor assessment and modification, staff education, gait assessment and intervention, assistive device assessment and optimization, as well as environmental assessment and modification. Although there is no association between the use of hip protectors and fall rates, their use should be encouraged because the ultimate goal of any fall prevention program is to prevent fall-related morbidity. PMID- 15530180 TI - Parkinson's disease without expected neuropathologic abnormality. AB - A case is reported of an initially 78-year-old man whose presentation and course, closely followed over 10 years by an academic neurologist, were consistent with classic idiopathic Parkinson's disease (PD), including unilateral onset, obvious cogwheeling, and a very good prolonged response to levodopa/carbidopa (LD/CD). Yet at autopsy, there was no neuronal loss in the substantia nigra nor were there any Lewy bodies or immunochemical evidence of alpha synuclein in the multiple brain structures studied. This case does not support the hypothesis that the use of LD/CD is toxic to the substantia nigra in people. This patient had been on traditional doses of LD/CD for approximately 10 years, yet the number of cells in the substantia nigra was well within the normal range at autopsy. These findings are not unique, but point out the need to explain the occurrence of typical PD symptoms and course in the absence of any PD-related neuropathologic changes. PMID- 15530181 TI - A quality improvement study: medication error leading to thyrotoxicosis and death. PMID- 15530182 TI - Home hospitalization service for acute uncomplicated first ischemic stroke in elderly patients. PMID- 15530184 TI - Medicare reform's impact on long-term care. PMID- 15530183 TI - Treating vitamin D deficiency in long-term care: it should be done yesterday, but how? PMID- 15530185 TI - Are current approaches to postfall assessment in nursing homes adequate? PMID- 15530186 TI - Asymmetric dimethyl arginine and symmetric dimethyl arginine levels in infants with persistent pulmonary hypertension of the newborn. AB - OBJECTIVE: We investigated whether infants with persistent pulmonary hypertension had elevated levels of asymmetric dimethyl arginine, an endogenous inhibitor of nitric oxide synthase, and symmetric dimethyl arginine, a regioisomer. DESIGN: Prospective observational cohort study. SETTING: A 10-bed neonatal intensive care unit in a tertiary referral center. PATIENTS: Forty five infants >34 wks gestation and <2 wks old admitted to our intensive care unit. INTERVENTIONS: Samples of urine on days 1, 3, and 5 were analyzed by high-performance liquid chromatography to determine asymmetric dimethyl arginine and symmetric dimethyl arginine levels. The clinical progression and treatment of the infants were noted. MEASUREMENTS AND MAIN RESULTS: Twenty-nine infants had a clinical diagnosis of persistent pulmonary hypertension confirmed on echocardiography, and there were 16 control infants. Median asymmetric dimethyl arginine levels on day 1 were significantly higher in the persistent pulmonary hypertension group (n = 29), 14.8 (10.3-21.7) mmol.mmol creatinine(-1).L(-1), compared with controls (n = 16), 3.6 (1.4-5.2) mmol.mmol creatinine(-1).L(-1) (p < .001). Asymmetric dimethyl arginine levels decreased to control levels by day 5 (p = .33). Symmetric dimethyl arginine levels were significantly higher than controls on day 1, 31.0 (21.7-65.9) vs. 14.7 (4.1-20.2) mmol.mmol creatinine(-1).L(-1) (p = .001) and day 3, 34.7(20.3-42.5) mmol.mmol creatinine(-1).L(-1) (p = .0001) and by day 5 had decreased significantly (p = .007) back to 16.7 (12.3-23.8) mmol.mmol creatinine( 1).L(-1), which was not significantly different than the control group values. CONCLUSIONS: These results support the hypothesis that asymmetric dimethyl arginine and symmetric dimethyl arginine levels are elevated in patients with persistent pulmonary hypertension. Thus, endogenous inhibition of nitric oxide synthase by asymmetric dimethyl arginine may be responsible for the development of persistent pulmonary hypertension, suggesting novel therapeutic options in persistent pulmonary hypertension. PMID- 15530187 TI - Sedation, analgesia, and neuromuscular blockade in the pediatric intensive care unit: survey of fellowship training programs. AB - OBJECTIVE: To survey current sedation, analgesia, and neuromuscular blockade practices in pediatric critical care fellowship training programs in the United States. DESIGN: Questionnaire survey sent by all program directors. The survey could be submitted either via a Web site, fax, or mail. SETTING: University school of medicine. SUBJECTS: Fifty-nine pediatric critical care fellowship training program directors in the United States, listed on the Accreditation Council for Graduate Medical Education Web site. INTERVENTIONS: Survey. MEASUREMENTS AND MAIN RESULTS: The response rate was 59.3% (35 questionnaires). Midazolam, lorazepam, morphine, and fentanyl are the most frequently used drugs in pediatric intensive care units for analgesia and sedation. Most pediatric intensive care units surveyed have a written sedation policy (66%). The majority of units responding to the survey (85.7%) routinely use a scoring system to assess agitation and pain in children, with the most common being the COMFORT score. All of the pediatric intensive care units surveyed reported weaning drugs slowly to try to prevent drug withdrawal. Movement disorders related to prolonged sedation and analgesia seem to be more common than is reported in the literature, with 65.7% of units reporting cases. There is good consensus on the indications for neuromuscular blockade, with vecuronium being the most popular drug. CONCLUSIONS: When compared with a similar survey from 1989, this survey suggests that pediatric critical care units with fellowship training programs have made some changes in their approach to sedation and analgesia over the past decade. More fellowship directors report the use of sedation protocols and better recognition, prevention, and management of drug withdrawal. Similar analgesic, sedative, and neuromuscular blocking drugs are being used but some more commonly than a decade ago. PMID- 15530188 TI - Increases in serum levels of troponin I are associated with cardiac dysfunction and disease severity in pediatric patients with septic shock. AB - OBJECTIVES: Myocardial cell injury may contribute to cardiac dysfunction in septic shock. Troponin I is a biochemical marker of myocardial cell injury and death. We hypothesized that troponin I is increased in pediatric patients with septic shock and correlates with cardiac dysfunction and disease severity. DESIGN: Prospective, observational study. SETTING: Children's medical center. PATIENTS: Twenty-three patients with septic shock and cardiovascular failure were enrolled. MEASUREMENTS AND MAIN RESULTS: Serum troponin I was measured at admission and serially over 72 hrs. Within 24 hrs of study enrollment, echocardiograms were performed to determine left ventricular ejection fraction, systolic fractional shortening, heart rate corrected mean velocity of circumferential fiber shortening, and end-systolic wall stress. Requirement for inotropic support (stratified as low, moderate, or high), number of organ system failures, and other demographic data (including Pediatric Risk of Mortality III) were collected. Troponin I was increased on admission in 13 of 23 patients (57%) and at 12 hrs in ten of 22 patients (46%). In all cases, troponin I was maximal within 12 hrs of admission. Admission troponin I was inversely correlated to ejection fraction and fractional shortening and directly correlated to wall stress. Patients who had increased admission troponin I had lower heart rate corrected mean velocity of circumferential fiber shortening (preload and heart rate independent measure of left ventricular systolic function) and higher wall stress (measure of afterload) compared with patients with normal troponin I. Admission troponin I correlated with Pediatric Risk of Mortality III and organ system failure but did not correlate with requirement for inotropic support. CONCLUSIONS: Troponin I was increased in >50% of septic children early in their illness. Increased admission troponin I was associated with decreased measures of systolic cardiac function, as measured by echocardiography, and correlated with severity of illness. Early myocardial cell injury may contribute to the development of subsequent organ failure in septic shock, and measuring troponin I on admission may be helpful in assessing severity of sepsis. PMID- 15530189 TI - Postoperative fluid balance influences the need for antihypertensive therapy following coarctation repair. AB - OBJECTIVE: The purpose of the investigation was to determine the effect of fluid management on the need for perioperative antihypertensive therapy following coarctation repair. DESIGN: Retrospective case study. SETTING: Pediatric intensive care unit. PATIENTS: Infants and children with repaired coarctation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Hypertension was defined as a systolic blood pressure exceeding the 95th percentile for age. Echocardiographic variables included pre- and postoperative coarctation gradients, shortening fraction, left ventricular wall stress, and velocity of circumferential shortening. Clinical variables included age, fluid input, urine output (0-72 hrs), estimated creatinine clearance, intensive care unit stay, and diuretic use. Twenty-four patients were identified and divided into two groups. Group 1 consisted of six normotensive patients (19%, 1 wk to 2 yrs) and group 2 included 18 patients (75%, 3 wks to 12 yrs) with hypertension who required antihypertensive therapy. Group 2 patients were older (37 months vs. 3 months), received more intraoperative fluid, had lower urine output with fewer patients receiving diuretics, had diuretic therapy started later, and had longer intensive care unit stays (p < .05). When compared with group 2, 83% of group 2 patients had a net positive fluid balance between 36 and 72 hrs postoperatively. There were no differences in mean pre-/postoperative coarctation gradients, systolic function, postoperative fluids, estimated creatinine clearance, or aortic cross clamp time. Using logistic regression analysis, we found that variables independently associated with the need for antihypertensive therapy included intraoperative fluid volume, 48- to 72-hr urine output, a positive fluid balance, and the use and timing diuretic therapy (p < .05). Subgroup analysis of infants <1 yr of age revealed similar findings. CONCLUSION: A net positive fluid balance caused by either the volume of intraoperative crystalloid infusion or a lower urine output contributes to the development of paradoxic hypertension following coarctation repair regardless of patient age. Limiting intraoperative fluids and early diuretic use may limit the need for antihypertensive therapy and shorten the intensive care unit stay. PMID- 15530190 TI - Experience with mobile inhaled nitric oxide during transport of neonates and children with respiratory insufficiency to an extracorporeal membrane oxygenation center. AB - OBJECTIVE: To share our experience with the use of inhaled nitric oxide (iNO) during the transport of ventilated neonates and children to an extracorporeal membrane oxygenation (ECMO) center and to discuss the efficacy and safety of iNO use in this situation. DATA SOURCES: Case note review of 55 consecutive patients transported while receiving iNO to Glenfield Hospital, Leicester, UK, for consideration of ECMO. STUDY SELECTION: Retrospective case note review. DATA EXTRACTION: The clinical condition of each patient recorded at arrival of the transport team at the referring hospital, during transport, and at arrival at Glenfield Hospital. Preclinical and postclinical conditions were compared using the paired Student's t-test. DATA SYNTHESIS: Overall data showed a significant improvement in transcutaneous oximetry measurements (Spo(2): 84.8% preclinical, 90.6% postclinical; p = .006) and Pao(2) (59 torr [7.87 kPa] preclinical, 84 torr [11.23 kPa] postclinical; p = .001) during transport in our patient group. Based on limited safety data, no untoward events or toxic metabolites were observed with iNO therapy during transport. CONCLUSIONS: iNO does appear to improve oxygenation during transfer of patients for ECMO in our series. Based on limited safety data, iNO appears safe to use in transport. PMID- 15530191 TI - Traumatic stress in parents of children admitted to the pediatric intensive care unit. AB - OBJECTIVE: To measure the prevalence of parental acute stress disorder (ASD) and posttraumatic stress disorder (PTSD) and to examine the relationship between ASD symptoms and PTSD symptoms in parents of infants and children admitted to the pediatric intensive care unit (PICU). To examine the correlation between parental perceptions of illness severity and objective measures. To assess the association among demographic, situational, and illness factors and the severity of ASD and PTSD. DESIGN: Prospective cohort study. SETTING: Thirty-eight bed PICU at an urban children's hospital. PATIENTS: The parents of 272 children admitted to the PICU for >48 hrs. INTERVENTIONS: ASD symptoms were assessed using the Acute Stress Disorder Scale during the child's admission. PTSD symptoms were assessed using the PTSD Checklist at least 2 months after discharge. The severity of illness was measured using the Pediatric Risk of Mortality (PRISM III) score. MEASUREMENTS AND MAIN RESULTS: Of the 272 parents completing the initial assessment, 87 (32%) met symptom criteria for ASD. Of the 161 parents completing follow-up, 33 (21%) met symptom criteria for PTSD. PTSD symptoms at follow-up were associated with ASD symptoms assessed in the PICU, unexpected admission, parent's degree of worry that the child might die, and the occurrence of another hospital admission or other traumatic event subsequent to the index admission. Neither ASD nor PTSD responses were associated with objective measures of a child's severity of illness (PRISM III score). CONCLUSION: Traumatic stress symptoms are common among parents in the PICU and may persist long after discharge. There is strong support from these data for continued attention to supporting parents both during and after a child's PICU admission. PMID- 15530192 TI - Heme oxygenase-1 messenger RNA expression is induced in peripheral blood mononuclear cells of pediatric cancer patients with systemic inflammatory response syndrome. AB - OBJECTIVE: To determine whether heme oxygenase-1 messenger RNA expression in peripheral blood mononuclear cells is induced in pediatric cancer patients with the systemic inflammatory response syndrome (SIRS) and whether this expression correlates with the heme oxygenase-1 products, bilirubin and carboxyhemoglobin. DESIGN: Prospective, controlled study. SETTING: A tertiary care pediatric oncology hospital. PATIENTS: Fourteen patients admitted to the intensive care unit with a diagnosis of SIRS by American College of Chest Physicians/Society for Critical Care Medicine consensus criteria and 17 control patients (off therapy, no acute illness). INTERVENTIONS: Blood for bilirubin, carboxyhemoglobin, and heme oxygenase-1 messenger RNA expression was collected at study entry. SIRS patients continued to have samples collected every 12 hrs for 1 wk or until intensive care unit discharge. Heme oxygenase-1, bilirubin, and carboxyhemoglobin levels of SIRS patients were compared with controls, and correlation between heme oxygenase-1 and products was assessed. MEASUREMENTS AND MAIN RESULTS: Within 48 hrs of study entry, maximum heme oxygenase-1 expression for all SIRS patients compared with controls was 5.5 +/- 1.0 vs. 1.1 +/- 0.1 (p < .0006). Maximum expression was > or =2.3-fold in 13 of 14 SIRS patients. Maximum heme oxygenase-1 expression also differed from minimum (5.5 +/- 1.0 vs. 1.6 +/- 0.3, p < .003). Maximum bilirubin and carboxyhemoglobin levels within 48 hrs of study entry differed between SIRS patients and controls (3.0 +/- 0.8 vs. 0.3 +/- 0.1, p = .006; and 1.2 +/- 0.2 vs. 0.6 +/- 0.1, p = .001, respectively). Bilirubin, but not carboxyhemoglobin, correlated with heme oxygenase-1 expression (p = .0013). CONCLUSIONS: Heme oxygenase-1 messenger RNA, bilirubin, and carboxyhemoglobin levels were increased within 48 hrs of admission in pediatric cancer patients with SIRS. Heme oxygenase-1 expression correlated with serum bilirubin levels. The increase in heme oxygenase-1 expression may add to the understanding of the increase in serum bilirubin observed in patients with SIRS/sepsis. These findings support a role for heme oxygenase-1 in the physiologic response to inflammatory stress. PMID- 15530193 TI - Randomized comparison between fluconazole and itraconazole for the treatment of candidemia in a pediatric intensive care unit: a preliminary study. AB - OBJECTIVE: Candida bloodstream infections have shown an increase in hospitalized patients, especially those receiving intensive care. The effectiveness of various azoles, especially itraconazole, in treatment of candidemia has not been fully evaluated. Our objective was to compare the efficacy and safety of enterally administered itraconazole vs. fluconazole in treatment of candidemia. DESIGN: Randomized, double-blind, controlled trial. SETTING: Pediatric intensive care unit of a referral and teaching hospital. SUBJECTS: Forty-three pediatric patients with candidemia, INTERVENTION: Patients received either fluconazole (n = 22) or itraconazole (n = 21), about 10 mg/kg orally or through a gastric tube, and were monitored for clinical and mycological cure (sterile fungal blood culture), blood counts, and liver and renal functions. MEASUREMENTS AND MAIN RESULTS: The clinical characteristics of two groups were comparable. The cure rate was similar in both the groups: itraconazole 17 of 21 (81%) and fluconazole 18 of 22 (82%). Crude mortality rate (itraconazole 9.5% and fluconazole 13.6%) was also comparable in two groups of patients. The frequency of electrolyte disturbance was very low and similar in both the groups. Blood urea, creatinine, liver enzymes, and serum bilirubin were not adversely affected. CONCLUSIONS: Itraconazole was as effective as fluconazole in nosocomial candidiasis in children receiving intensive care and was devoid of serious side effects. PMID- 15530194 TI - The influence of timing of elective cesarean section on neonatal resuscitation risk. AB - OBJECTIVE: Cesarean section has negative effects on the physiologic responses to birth, including the development of lung volumes, pulmonary vascular resistance, and biochemical responses. The objective of this study was to examine the association between the timing of delivery between 37 and 42 wks gestation and neonatal resuscitation risk in elective cesarean section. DESIGN: Observational, cohort study. SETTING: Maternity Department of Padua University, Italy. SUBJECTS: All pregnant women who delivered by elective cesarean section at term during a 3 yr period were identified from a perinatal database and compared retrospectively with pregnant women who delivered vaginally and matched for week of gestation. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Comparative neonatal resuscitation risk (odds ratio, OR; confidence interval, CI) was analyzed. During this time, 1,284 (13%) elective cesarean section deliveries occurred at or after 37 wks of gestation. Forty-four (3.4%) newborns delivered by elective cesarean section and 18 (1.4%) newborns vaginally delivered needed positive pressure ventilation resuscitation by laryngeal mask airway or tracheal tube. Positive pressure ventilation resuscitation risk was significantly higher in the infant group delivered by elective cesarean section compared with vaginal delivery (OR, 2.05; CI, 1.25-5.67; p < .01) and involved both laryngeal mask airway and tracheal tube resuscitation maneuvers (OR, 2.77 CI, 1.26-5.8; p < .01 and OR, 2.9; CI, 1.02-7.81; p < .01, respectively). In the period of weeks 37(+0) to 38(+6), positive pressure ventilation resuscitation risk and single laryngeal mask airway and tracheal tube resuscitation maneuver risk were significantly greatly increased (OR, 4.25; CI, 1.46-16.12; p < .01; OR, 2.25; CI, 1.46-6.12; p < .01; and OR, 11.3; CI, 2.15-16.0; p < .01, respectively). After 38(+6) weeks, there was no significant difference in positive pressure ventilation resuscitation risk. CONCLUSIONS: Elective cesarean section at term, in an obstetric population without prenatally identified risk factors, remains associated with increased resuscitation risk with related implications for the neonate compared with vaginal delivery. A significant reduction in neonatal resuscitation risk would be obtained by waiting until week 39(+0) before performing elective cesarean section. PMID- 15530196 TI - Hashimoto's encephalopathy: an unusual cause of seizures in the intensive care unit. AB - OBJECTIVE: To present a case study of Hashimoto's encephalopathy. DESIGN: Case report. SETTING: Paediatric Intensive Care Unit, Nottingham University Hospital, UK. PATIENTS: Two adolescent females presented with encephalopathy and raised venous lactate. Both had subtle signs of neurocognitive deterioration before initial presentation. Extensive investigation revealed elevated antithyroid antibody titer, suggesting Hashimoto's encephalopathy. INTERVENTIONS: Steroid administration. MEASUREMENTS AND MAIN RESULTS: Symptoms rapidly resolved in both cases after steroid treatment. CONCLUSION: Hashimoto's encephalopathy should be considered in cases of unexplained encephalopathy presenting to the intensive care unit. Teenage girls with an antecedent history suggestive of thyroid disease or progressive cognitive decline warrant special attention. Antithyroid antibody titers should be measured even if standard thyroid function tests are normal. Although the etiology is unknown, prompt steroid responsiveness suggests an inflammatory or autoimmune disorder, and patients should be treated accordingly. PMID- 15530195 TI - Tezosentan, a combined parenteral endothelin receptor antagonist, produces pulmonary vasodilation in lambs with acute and chronic pulmonary hypertension. AB - OBJECTIVE: To investigate the hemodynamic effects of tezosentan in the intact lamb both at rest and during acute and chronic pulmonary hypertension. DESIGN: Prospective, randomized experimental study. SETTING: University-based research laboratory. SUBJECTS: Lambs with and without pulmonary hypertension. INTERVENTIONS: Six newborn lambs were instrumented to measure vascular pressures and left pulmonary blood flow. The hemodynamic effects of tezosentan (0.5, 1.0, 5.0 mg/kg, intravenously) were studied at rest and during U46619-induced pulmonary hypertension. Following in utero placement of an aortopulmonary vascular graft, nine additional lambs with increased pulmonary blood flow and chronic pulmonary hypertension (shunt) were also studied at 1 wk (n = 5) and 8 wks (n = 4) of age. MEASUREMENTS AND MAIN RESULTS: At rest, tezosentan had no significant effect on any of the variables. During acute U46619-induced pulmonary hypertension, tezosentan caused a dose-dependent decrease in pulmonary arterial pressure (from 5.9% +/- 4.7 to 16.0% +/- 10.7; p < .05) and pulmonary vascular resistance (from 6.2% +/- 8.0 to 21% +/- 8.8; p < .05). Mean systemic arterial pressure was unchanged. In 1- and 8-wk-old shunt lambs with increased pulmonary blood flow, tezosentan (1 mg/kg) produced potent nonselective pulmonary vasodilation. CONCLUSIONS: Tezosentan, a combined endothelin receptor antagonist optimized for parenteral use, induces potent selective pulmonary vasodilation during acute U46619-induced pulmonary hypertension and potent nonselective vasodilation in chronic pulmonary hypertension secondary to increased pulmonary blood flow. In general, the hemodynamic effects of bolus doses of tezosentan occurred within 60 secs of administration and lasted approximately 5-10 mins. The hemodynamic profile of intravenous tezosentan may make it a useful adjunct therapy for acute pulmonary hypertensive disorders and warrants further study. PMID- 15530197 TI - First Pediatric Intensive Care Congress in Chile. AB - OBJECTIVE: To summarize the First Chilean Congress on Pediatric Critical Care Medicine. DATA SOURCE/EXTRACTION: Program schedule, presentations, abstracts. STUDY SELECTION: Descriptive summary of the congress. CONCLUSIONS: The Chilean Society of Pediatric Critical Care organized the First Pediatric Intensive Care Congress in Chile, having a registration of 217 delegates. All the pediatric intensive care units of the country had a representative in the Congress either as a delegate or a faculty member, which provided an excellent opportunity for interactions between them. The international faculty addressed state-of-the-art issues, with a charge of highlighting basic science relevant to clinical practice, which motivated long interactive question-and-answer sessions with the audience. Strong evidence was presented by Chilean faculty debaters who discussed controversial topics, making these sessions very attractive. An entire day was scheduled with a simultaneous nursing session. More than 80 intensive care nurses benefited from a very interactive day with five symposia and short oral presentations. A randomized multi-institutional research protocol studying reintubation rate using three methods for weaning from mechanical ventilation was discussed during the meeting. Thirteen units committed to participate. Twenty three posters were displayed in the congress. Posters describing the activity of pediatric intensive care units were also presented. The mean for intensive care unit beds was 6.8 and for intermediate care beds was 7.1. Annual admissions showed a mean of 640 patients, and mortality rate ranged from 4.1% to 12%, with a mean of 6.7% PMID- 15530198 TI - The interactive effects of prior knowledge and text structure on memory for cognitive psychology texts. AB - BACKGROUND: Interest in the interactive effects of prior knowledge and text structure on learning from text is increasing but experimental manipulations of knowledge and structure variables often produce findings that do not help teachers to select expository texts for students. AIMS: We aimed to extend the ecological validity of previous findings by asking students with a high or low level of discipline-relevant knowledge to read texts characteristic of those they would normally encounter. A compensation effect was hypothesized, where high prior knowledge would compensate for a lack of text structure and text structure would compensate for a lack of prior knowledge. SAMPLES: One hundred and ninety five undergraduate psychology students (144 Year 1 students and 51 Year 3 students) were allocated to a high knowledge (HiPK) or low knowledge (LoPK) group on the basis of their performance on a word association test. METHODS: Participants were randomly assigned to one of five text structure groups (compare/contrast, sequence, classification, enumeration, generalization) and asked to study two cognitive psychology passages before recalling the main points of the text immediately afterwards and after a delay of 2 weeks. The five text structures were placed on an 'organizational continuum' according to the degree of structure/organization in the 10 passages. RESULTS: A compensation effect did not emerge. Recall was high when texts were well structured and readers had prior knowledge, but recall was poor when texts were less structured, regardless of the level of prior knowledge. CONCLUSIONS: Readers benefit most from texts that challenge pre-existing mental representations. PMID- 15530199 TI - Does the cue help? Children's understanding of multiplicative concepts in different problem contexts. AB - BACKGROUND: Understanding arithmetical principles is a key part of a conceptual understanding of mathematics. However, very little attention has been paid to children's understanding of multiplicative, as compared to additive, principles. AIMS: This study investigated (a) children's ability to use commutative and distributive cues to solve multiplication problems, (b) whether their ability to use these cues depends on the problem context, and (c) whether separate mechanisms might underlie children's understanding of commutativity and distributivity. SAMPLE: Twenty-seven 9-year-olds (Year 5) and thirty-two 10-year olds (Year 6). METHODS: Forty-eight multiplication problems (with a multiple choice response format) were presented to children. There were four types of problem: Commutative, Distributive, Combined commutative-distributive (all preceded by a cue) and No cue problems. Each type of problem was presented in three different contexts: Isomorphism of measures, Area, and Cartesian product. RESULTS: Children demonstrated a good understanding of commutativity but a very poor understanding of distributivity. A common mistake in the distributive problems was to select the number that was one more, or one less, than the answer in the cue. Children's understanding of distributivity (but not commutativity) seemed to depend on the problem context. Factor analysis suggested that separate factors underlie the ability to solve commutative and distributive problems. CONCLUSIONS: Nine- and 10-year-olds understand commutativity, but are unable to use the distributive principle in multiplication. Their errors suggest that they may confuse some of the principles of multiplication with those of addition. When children do begin to understand the principle of distributivity, they most easily apply it in the context of Isomorphism of measures multiplication problems. The implications for mathematical education are discussed. PMID- 15530200 TI - Orthographic revision: a developmental study of how revisers check verbal agreements in written texts. AB - BACKGROUND: Writing is a complex activity involving various cognitive processes in the planning, the transcription and the revision of written texts. The present study focused on the revision of written texts within a developmental approach. AIMS: The study aimed to examine whether children and adults use different procedures to detect and revise erroneous grammatical agreements. It was predicted that children would use a slow algorithmic procedure while adults would use a fast automatized procedure. SAMPLE: One hundred and twenty participants from 5th grade to undergraduate levels (24 per level) participated in the study. METHOD: The participants were asked to decide as quickly as possible whether a visually presented sentence had any agreement error. The French experimental sentences were of the type 'The N1 of the N2 + Verb', in which N2 was either a plausible subject of the following verb (e. g., The guard of the prisoners watches) or an implausible subject (e. g., The guard of the safes watches). Correctness and latency of the responses were recorded. RESULTS: The main results showed that only the younger participants were affected by the subject-role plausibility of N2, and that there was no difference in response latency between their correct and incorrect responses. These observations support the hypothesis that the younger participants systematically apply a time-consuming algorithmic procedure to verify the agreement; since one step of this procedure consists in searching for the subject of the verb, these participants were frequently misled by the subject-role plausibility of N2. On the contrary, the older participants were not affected by the plausibility of N2, but were frequently misled by erroneous agreements between N2 and the verb. These observations support the view that these older participants use a fast decision strategy based on the co occurrence of formal indices. Their correct answers, however, were slower than their incorrect ones; this suggests that they also sometimes use a time-consuming controlled procedure. CONCLUSION: The study shows that along with the acquisition of writing expertise, the revising activity itself is progressively facilitated and gradually automatized by substituting a fast direct decision strategy for a slow and laborious use of revision rules. PMID- 15530201 TI - Relationship between orthographic-motor integration and computer use for the production of creative and well-structured written text. AB - BACKGROUND: Orthographic-motor integration refers to the way in which orthographic knowledge is integrated with fine-motor demands of handwriting. A strong relationship has shown to exist between orthographic-motor integration and students' ability to produce creative and well-structured written text (De La Paz & Graham, 1995). This relationship is thought to be due to the cognitive load resulting from lack of automaticity in orthographic-motor integration so that writers do not have sufficient resources to accomplish the more demanding aspects of writing. Interventions to improve children's orthographic-motor integration result in improved written text (Jones & Christensen, 1999). AIM: This study first extended findings related to handwritten text to the relationship between typing and the length and quality of computer-based written text. Second, it examined the efficacy of an intervention to develop proficiency in typing skills on the length and quality of students' written language. SAMPLE: Participants in the first study were 276 Grade 8 and 9 students. In the second study 35 students in Grades 8 and 9 who exhibited very low levels of proficiency in typing were the participants. METHODS: In Study 1, orthographic-motor integration related to typing as well as handwriting was assessed for all students. They were asked to complete a piece of handwritten and computer-based text. Students in the intervention study completed the same measures as Study 1, at pre- and post-test. During the intervention half the students completed a daily typed journal and half completed a program designed to facilitate their typing skills. RESULTS: There was a significant relationship between orthographic-motor integration -- handwriting and the length and quality of handwritten text, and a stronger relationship between orthographic-motor integration -- typing and length and quality of computer-based text. Both intervention groups in the second study showed significant differences in writing skills from pre- to post-test. However, the typing skills group showed significantly better scores on typing and quality of typewritten text than the journal group at post-test. The impact of the intervention was specific to typewritten text. There was no difference in length or quality of handwritten text. CONCLUSION: It is suggested that developing proficiency in orthographic-motor integration related to typing allows writers to employ their cognitive resources more flexibly when working on a computer, so that they can devote attention to higher-order processes involved in ideation, syntactic and semantic monitoring and pragmatic awareness. PMID- 15530202 TI - Profiles of non-victims, escaped victims, continuing victims and new victims of school bullying. AB - BACKGROUND: Victims of school bullying are known to be at risk in peer relationships and to sometimes use ineffective coping strategies, but little previous research has examined differences among escaped victims, continuing victims and new victims. AIM: A follow-up design compared friendships, behavioural characteristics, victimization experiences and coping strategies of pupils who had 2 years previously answered a questionnaire identifying themselves as victims (V) or non-victims (NV) of school bullying and whose current victim status could be identified. SAMPLE: 406 pupils aged 13-16 years (190 boys, 216 girls): 175 non-victims (NV-NV), 146 escaped victims (V-NV), 27 new victims (NV V) and 58 continuing victims (V-V). METHOD: Structured interviews were given to pupils, together with the SDQ. Teachers also filled in the SDQ. School records of attendance were obtained. RESULTS: Escaped victims did not differ greatly from non-victims, but had some self-perception of continuing peer relationship difficulties. Continuing victims, irrespective of gender, liked other pupils and breaktime less (but did not dislike other aspects of school), had fewer friends in school (but not outside school), more often missed school (sometimes because of bullying), scored high on problem scales of the SDQ, and were more likely to be involved in bullying others as well as being bullied. New victims tended to resemble continuing victims. Continuing victims did not differ from escaped victims on type of bullying, but new and continuing victims less often reported talking to someone about a specific incident of bullying. Most victims gave mainly victim-related reasons for the bullying having taken place. CONCLUSION: The results are discussed in relation to why some pupils become or continue to be victims in secondary school, and recommendations for anti-bullying procedures in schools designed to help such victims. PMID- 15530203 TI - 'What about bullying?' An experimental field study to understand students' attitudes towards bullying and victimization in Italian middle schools. AB - BACKGROUND: Attitudes towards bullying at school are influential in understanding and preventing bullying behaviour but they should be measured with reference to the particular conditions under which bullying takes place. AIMS: To establish how far positive and negative judgments of bullying and victims and blaming of the victim vary according to the gender of observers, gender of bullies and of victims and whether the bullying took place alone or in group. SAMPLE: Participants were 117 students (49 boys and 68 girls), aged 11-12, recruited from a middle school in Italy randomly allocated to one of four independent groups according to experimental condition: bullying alone among girls, bullying alone among boys, bullying in groups among girls, bullying in groups among boys. METHOD: Participants watched one of four versions of a video according to experimental condition showing a brief standardized bullying episode taking place at a school; they then had to fill in a self-report questionnaire measuring the dependent variables: respondents' positive or negative judgments towards the bully and the victim shown in the video and how far the victim was blamed for what had happened. RESULTS: Overall, results indicate students have positive attitudes towards the victims of bullying and tend not to blame them for what has happened. However, same gender identification lead girls to blame male victims more than female victims and the reverse applies in case of boys providing their judgments. A bully acting alone is considered stronger and braver than when acting in a group. CONCLUSIONS: The limits and potential of the study are presented with special attention to implications for intervention strategies in school by focusing on the role observers could play in supporting the victims and discouraging the bullies. PMID- 15530204 TI - The impact of supervisors' cognitive styles on the quality of research supervision in management education. AB - BACKGROUND: An aspect of teaching and learning that has been seriously overlooked in higher education is the process of research supervision. High failure rates for research dissertations in the social sciences have been partly attributed to student dissatisfaction with supervision and poor student-supervisor relationships. One personality variable that has been shown to be partly responsible for shaping the effectiveness of supervisory relationships is cognitive style. AIMS: The study examined the effects of supervisor cognitive style on the quality of supervision for students undertaking a research project in the field of management education. SAMPLE: Both parties in each of 118 supervisor-student dyads within a university business school in the UK participated in the study. METHOD: Data were collected using the Cognitive Style Index to measure subjects on the analytic-intuitive dimension of cognitive style. A self-developed Thurstone attitude scale was used to measure students' perceptions of the quality of supervision. The scale's validity was assured by making extensive use of subjects' (N = 100) judgments from the population of interest in the scale's development. A second parallel scale was developed to test the instrument's reliability characteristics. RESULTS: Findings revealed that students perceived the quality of supervision to increase significantly with the degree to which supervisors were analytic in their cognitive style. Students whose supervisors were more analytic also achieved significantly higher grades for their dissertations. CONCLUSIONS: Whilst there may be many factors influencing interpersonal relationships of this nature, this study demonstrated the potential relevance of cognitive style, which may prove to be a fertile area for further investigation. PMID- 15530205 TI - Consistency in reasoning test scores over time. AB - BACKGROUND: UK schools have a long history of using reasoning tests, most frequently of Verbal Reasoning (VR), Non-Verbal Reasoning (NVR), and to a lesser extent Quantitative Reasoning (QR). Results are used for identifying students' learning needs, for grouping students, for identifying underachievement, and for providing indicators of future academic performance. Despite this widespread use there are little empirical data on the long-term consistency of VR, QR and NVR as discrete abilities. AIMS: To evaluate and compare the consistency of VR, QR and NVR scores over a 3-year period, and to explore the influence of the secondary school on pupils' progress in the tests. SAMPLE: Data were collected on a longitudinal sample of over 10,000 pupils who completed the Cognitive Abilities Test Second Edition in year 6 (age 10+) and year 9 (age 13+), and GCSE public examinations in year 11 (age 15+). METHODS: Correlation coefficients and change scores for individual pupils are calculated. Multilevel modelling is used to determine school effects on reasoning scores and GCSE public examination results. RESULTS: The results reveal high correlations in scores over time, ranging from .87 for VR to .76 for NVR, but also show around one-sixth of pupils on the VR test and one-fifth of pupils on the QR and NVR tests change their scores by 10 or more standard score points. Schools account for only a small part of the total variation in reasoning score, although they account for a much greater proportion of the variation in measures of attainment such as GCSE. School effects on pupils' progress in the reasoning tests between age 10 and age 13 are relatively modest. CONCLUSIONS: Reasoning tests make excellent baseline assessments for secondary schools. Some practical and policy implications for schools are discussed. PMID- 15530206 TI - Illness representations in depression. AB - BACKGROUND AND OBJECTIVES: Illness representations in physical health problems have been studied extensively using the Self-regulation Model (SRM) focusing on five dimensions of illness beliefs (identity, consequences, causes, timeline and control, or cure). Associations have been found between beliefs about illness and a range of health outcomes. This study aimed to examine models of depression, to assess whether the five dimensions of the SRM are relevant, to compare depression models with those for physical illness, and to examine the psychometric properties of the Illness Perception Questionnaire (IPQ) when used with depression. DESIGN AND METHOD: A sample of 101 women either currently depressed or with a history of depression was asked to write about their experiences of physical sickness and depression. Their responses were analysed in terms of the dimensions of beliefs expressed and the two experiences were compared. The IPQ was also administered to assess the women's perceptions of depression. RESULTS: The women used the same five dimensions of illness as identified in the SRM in describing both their experience of depression and physical sickness. There was evidence of some consistency across the models of the two illnesses in terms of their content and structure. The IPQ was a reliable measure for depressed experiences and discriminated between women who were currently depressed or not. Comparing the women's descriptions of their depression with their IPQ scores showed some relationships between their responses on the two different measures, at least for the consequences and cause dimensions. CONCLUSION: The SRM model and associated methodology may provide an appropriate framework to further explore illness representations in depression. Problems inherent in the study of illness models in depression including the influence of mood on the model are described. Applications of this research area to the understanding of treatment preferences and adherence to treatment in mood disorders are discussed. PMID- 15530208 TI - Overgenerality of autobiographical memory in Alzheimer's disease. AB - OBJECTIVES: Overgenerality of autobiographical memory (ABM) is well documented in a range of clinical conditions, particularly in depressed and parasuicidal patients (e.g., Williams, 1996; Williams & Broadbent, 1986). This study extended the investigation to Alzheimer's disease (AD), and attempted to identify whether ABM overgenerality in the AD group is specifically expressed through an excess of categoric memories. DESIGN: AD sufferers and control participants were compared on their ABM specificity in a cued-recall task. METHOD: Ten AD patients and 10 controls, matched for age, gender and educational level, were administered an ABM specificity measure following their mental status assessment. A battery of neuropsychological tests provided an independent estimate of cognitive deficit severity in the following areas: long-term memory, IQ, working memory, processing speed, and verbal fluency. A control for depression was also employed. RESULTS: Compared to control participants, AD participants produced significantly fewer specific autobiographical memories. Additionally, the number of produced categoric overgeneral memories was significantly greater in the AD group in comparison with the controls. CONCLUSION: This study demonstrates the existence of ABM overgenerality in AD, manifested through an excess of categoric memories. Consistent with the mnemonic interlock theory (Williams, 1996), AD sufferers seem to lack cognitive resources to conduct a directed search for a specific memory, and stop at the categoric descriptions stage. This may contribute to lack of specificity in ABM retrieval. PMID- 15530207 TI - The role of overgeneral autobiographical memory in the development of adult depression following childhood trauma. AB - OBJECTIVE: Overgeneral autobiographical memory (AM) has been associated with episodes of clinical depression in adults and also with reported experience of childhood sexual abuse (CSA). This study investigated whether AM has a role in the development of adult depression in survivors of CSA and whether it is related to circumstances of CSA. METHOD: A community sample of women who reported a history of CSA (N = 41) completed the autobiographical memory test and were interviewed about any adult episodes of depression using the Schedule for Affective Disorders and Schizophrenia-Lifetime. RESULTS: Women who reported episodes of depression recalled fewer categoric memories in response to cue words than those who reported no episodes of depression. Correlations indicated that the number of categoric memories recalled for all cue words was associated with early abuse and greater duration of abuse. CONCLUSIONS: Relationships between AM and the age, and duration of abuse are consistent with previous findings of a relationship between CSA and AM. Categoric AM appears to serve as a defence against distressing memories. However, this suggestion needs to be tested with prospective studies showing whether AM recall style becomes more specific as a result of an episode of depression. PMID- 15530209 TI - Personality characteristics in obsessive-compulsive disorder and individuals with subclinical obsessive-compulsive problems. AB - OBJECTIVE: To assess the relationship between normal personality traits and obsessive-compulsive (OC) phenomena in individuals with subclinical OC problems and patients whose problems met diagnostic criteria for obsessive-compulsive disorder (OCD). METHOD: In Study 1, 25 healthy volunteers with high scores on the Padua Inventory (PI) and 28 controls with low scores on the PI were compared on the Sensitivity to Punishment and Sensitivity to Reward Questionnaire (SPSRQ), the Eysenck Personality Questionnaire (EPQ), and measures of depression and state anxiety. In Study 2, 56 treatment-seeking participants meeting DSM-IV criteria for OCD and 40 healthy volunteers of similar sociodemographic characteristics were compared on the same measures. RESULTS: Both individuals with subclinical OC problems and OCD patients scored significantly higher than their respective control groups on sensitivity to punishment, neuroticism and psychoticism. OCD patients, but not individuals with subclinical OC problems, scored lower in extraversion than their respective controls. Neuroticism was the strongest predictor of high scores on the PI in Study 1, while psychoticism was the strongest predictor of the presence of an OCD diagnosis in Study 2. CONCLUSION: Healthy participants with high scores on OC measures and OCD patients share various personality traits but can also be distinguished according to the level of extraversion, neuroticism and psychoticism. PMID- 15530210 TI - Thought-shape fusion in eating disorders. AB - OBJECTIVES: The aims of the present study were (1) to examine the association between a cognitive distortion ('thought-shape fusion') and eating disorder psychopathology and (2) to examine the degree of thought-shape fusion in people with eating disorders and a non-eating-disorder control group. DESIGN: Associations between thought-shape fusion and eating disorder psychopathology were examined and the degree of thought-shape fusion was compared between people with and without clinical eating disorders. METHOD: Forty-two women with clinical eating disorders and a group of 42 age-matched women with no self-reported history of an eating disorder completed self-report questionnaires to assess thought-shape fusion, eating disorder symptoms, body checking and body avoidance, and depression. RESULTS: Thought-shape fusion was significantly associated with eating disorder psychopathology. The majority of the associations remained significant when controlling for levels of depression. Patients with eating disorders showed significantly more thought-shape fusion than the non-clinical controls. CONCLUSIONS: Thought-shape fusion is a cognitive distortion associated with eating disorders. It may be a direct expression of the overevaluation of eating, shape and weight. It is recommended that thought-shape fusion be tackled directly in cases where it is a barrier to changing the core psychopathology of eating disorders. PMID- 15530211 TI - Development and preliminary validation of the Glasgow Content of Thoughts Inventory (GCTI): a new measure for the assessment of pre-sleep cognitive activity. AB - OBJECTIVE: To develop a self-report measure (the Glasgow Content of Thoughts Inventory [GCTI]) for the assessment of pre-sleep cognitive activity in adults with sleep-onset insomnia. DESIGN: A psychometric, scale development approach was used. METHOD: Over three consecutive nights, 12 people with insomnia provided 'live' audio-recordings of pre-sleep thought content, which were used to generate an item pool. The results were compared to the content and categorical structure of pre-sleep cognitive activity identified by Wicklow and Espie (2000), and commonalities in thought content were used to generate a draft scale. Following further piloting, a 25-item scale was developed and administered to two groups (29 people with insomnia and 29 good sleepers), along with other self-report measures, objective (actigraphic recordings) and subjective (diary) sleep indices, and results analysed to evaluate the psychometric properties of the scale. RESULTS: The GCTI demonstrated evidence of construct validity, successfully discriminated between individuals with insomnia and good sleepers, and was significantly correlated with existing measures of sleep disturbance. A score of 42 yielded a sensitivity of 100% and specificity of 83%. The GCTI demonstrated good test- retest reliability (ICC = .88) and internal consistency (alpha = .87). CONCLUSIONS: The GCTI appears to be a valid and reliable instrument for use with patients with sleep-onset insomnia. PMID- 15530212 TI - Systematic review of the efficacy of cognitive behaviour therapies for childhood and adolescent anxiety disorders. AB - PURPOSE: To review the effectiveness of cognitive behaviour therapy (CBT) as a treatment for anxiety disorders of childhood and adolescence. METHOD: Studies were included if they treated young people (under 19 yrs) with diagnosed anxiety disorder (excluding trials solely treating phobia, PTSD or OCD), had a no treatment control group, and used diagnosis as an outcome variable. A search of the literature, incorporating electronic databases, hand search and expert consultation, yielded 10 randomized controlled trials that were appropriate for inclusion. RESULTS: The outcome of interest was remission of anxiety disorder. Employing conservative criteria, the remission rate in the CBT groups (56.5%) was higher than that in the control groups (34.8%). The pooled odds ratio was 3.3 (CI = 1.9-5.6), suggesting that CBT has a significant effect. CONCLUSIONS: CBT is useful for the treatment of anxiety in children over the age of 6 years. However, we still know little about the treatment of younger children or about the comparative efficacy of alternative treatments. Most of the trials were efficacy trials, and have limited generalizability. Reporting of many aspects of the trials was weak. PMID- 15530213 TI - The self-report version of the Strengths and Difficulties Questionnaire: its psychometric properties in 8- to 13-year-old non-clinical children. AB - OBJECTIVE: To examine the reliability and validity of the self-report version of the Strengths and Difficulties Questionnaire (SDQ) in relatively young children. METHOD: The SDQ was administered to a large sample of non-clinical children (n = 1111) aged 8-13 years. In a subsample (n = 439), self-report SDQ scores of children with and without behaviour problems were compared, and related to the teacher version of the SDQ, the Youth Self-Report and the Teacher Report Form. RESULTS: Although the reliability of the self-report SDQ was somewhat less satisfactory in the younger children of our sample, most other psychometric properties were acceptable and comparable to those obtained in older youths. CONCLUSION: While the self-report SDQ was designed for youths aged 11 years and above, the current data seem to suggest that the scale may provide useful information about psychopathological symptoms in children as young as 8 years old. PMID- 15530214 TI - Parent training for Attention Deficit/Hyperactivity Disorder: is it as effective when delivered as routine rather than as specialist care? AB - BACKGROUND: The effectiveness of parent training (PT) when delivered as part of specialist tier-two services for preschool AD/HD children has been recently demonstrated. AIMS: To assess the effectiveness of the same PT programme when delivered as part of routine primary care by non-specialist nurses. METHOD: A sample of 89 3-year-old children with preschool AD/HD took part in a controlled trial of an eight-week (one hour a week), health visitor delivered, PT package. Children, allocated randomly to PT (n = 59) and waiting list control (WLC; n = 30) groups, were compared. RESULTS: PT did not reduce AD/HD symptoms. Maternal well-being decreased in both PT and WLC groups. CONCLUSIONS: While PT is an effective intervention for preschool AD/HD when delivered in specialized settings, these benefits do not appear to generalize when programme are delivered as part of routine primary care by non-specialist nurses. PMID- 15530215 TI - Brief cognitive therapy of nightmares and post-traumatic ruminations in a man with a learning disability. AB - OBJECTIVES: This is a case study of the treatment of two recurrent nightmares, one of which was accompanied by congruent post-traumatic daytime ruminations, in a man with a mild to moderate learning disability. METHOD: Treatment was by a cognitive intervention in which the patient was taught new positive endings to his dream stories. RESULTS: Both of the nightmares, as well as the post-traumatic ruminations, responded to treatment, and gains were maintained at 12-month follow up. CONCLUSIONS: Cognitive treatment of nightmares can be successful even in a patient with very limited intellectual abilities. The discussion considers the factors that may have been responsible for the success of the intervention in a man who would not be considered an obvious candidate for cognitive therapy. PMID- 15530216 TI - Implications of the growing obesity epidemic on contraception and reproductive health. PMID- 15530217 TI - Human papillomavirus (HPV) vaccines: prospects for eradicating cervical cancer. PMID- 15530218 TI - Teenagers' use of sexual health services: perceived need, knowledge and ability to access. AB - INTRODUCTION: An individual teenager's use of services may depend on perceived need, on knowledge of sexual health and local services, and on ability to access. This paper presents the first UK large-scale quantitative analysis of these factors, comparing those who use services with those who do not. METHODS: 15/16 year-olds (n = 5747) were questioned about their use of sexual health services in the SHARE trial of a school sex education programme in 25 schools in Lothian and Tayside, Scotland, UK. Multilevel statistical models examined the role of different factors on service use. RESULTS: One-third of teenagers had used a service, and use was strongly related to sexual experience. In addition, some family influences and being a school leaver were associated with service use, although we found no evidence for class, ethnic or religious barriers to use. Proximity to specialist clinics was linked with greater use, while low spending money and high parental monitoring were associated with less use. Teenagers with better knowledge, who rated their school sex education as effective, who were comfortable talking about sex and who had discussed contraception with peers were more likely to have used services. Differences in use relating to sexual experience, knowledge, feeling comfortable talking about sex and talking with peers helped to explain gender differences in service uptake. CONCLUSION: There is potential to influence service use through better knowledge and confidence imparted through school sex education, and by improving the links between services and schools. PMID- 15530219 TI - Community attitudes to the sexual behaviour of young people in an urban area with high rates of sexual ill-health. AB - CONTEXT: Adults in any community are a potentially important source of sexual health information for young people. Open discussion of sexual health issues is associated with low rates of sexual ill-health. Adults who disapprove of teenage sexual behaviour are poor sources of advice. The study of adult attitudes to the sexual behaviour of young people is relevant to work on improving access to sexual health services. SETTING: Adults' attitudes to the sexual behaviour of young people in an urban area with high indices of sexual ill-health were documented. DESIGN: Data were collected via questionnaires administered in popular shopping areas by local people after training. RESULTS: A total of 283 interviews were completed. Eighty-eight percent of respondents thought that the likely age of first sex among young people was under the age of consent but only 8% thought that the acceptable age of first intercourse was under 16 years. Knowledge of local services was suboptimal. Twenty percent of respondents did not know where young people could get contraception or advice on sexual health issues. Less than half (42%) suggested a general practitioner with a similar proportion suggesting a family planning clinic (FPC) or Brook clinic. When asked what services FPCs provide, only 40% mentioned contraception and 32% did not know. Despite their lack of knowledge, the majority (84%) of respondents would tell a young person where they could obtain contraception or sexual health advice. Seventy-six percent thought parents and 56% thought schools are the key sources of sexual health information for young people. CONCLUSIONS: Adults resident in this area have negative attitudes to the sexual behaviour of local young people and suboptimal knowledge of local contraceptive services. They do, however, identify themselves as potentially important sources of sexual health advice and may therefore benefit from more information and an opportunity to discuss their attitudes. PMID- 15530221 TI - FFPRHC Guidance (October 2004) contraceptive choices for young people. PMID- 15530220 TI - Unanswered questions in contraceptive management: what do the experts do? AB - CONTEXT: Several areas exist in the practice of contraception where evidence for practice is deficient, yet clinical decisions need to be made. OBJECTIVES: The aim of the study was to find the practice habits of lead practitioners in the area of contraception in specific clinical scenarios where the published evidence is inadequate to provide clear guidance to clinicians. Results can provide 'Level V' evidence for practice for the 'nonexpert' practitioner. DESIGN: Descriptive study. PARTICIPANTS: The study was conducted as a postal questionnaire mailed to the 205 lead practitioners whose contact details were known through the Society of Consultants in Reproductive Health (hereafter referred to as 'consultants') working in reproductive health in the National Health Service. RESULTS: A total of 138 consultants returned completed questionnaires (67% response rate). Important results included 100% of respondents being prepared to prescribe progestogen-only emergency contraception more than once in a cycle (contrary to product labelling) and 71% recommending two tablets daily of the progestogen-only pill for women of high body mass. CONCLUSIONS: Some questions had responses that showed clear majorities, providing a clear guide to practice, while other areas remain doubtful. Comments from respondents indicated great interest in all areas covered and a desire for consensus on many of the issues. Certainly the licensing and the advice from pharmaceutical companies is conservative, and in many scenarios a majority of consultants indicated that in order to serve the best interests of their clients they feel constrained to practise outside the Summary of Product Characteristics. PMID- 15530223 TI - The members' enquiry service: frequently asked questions. PMID- 15530224 TI - Nurse prescribing for contraceptive care and sexual health. PMID- 15530225 TI - The quality of qualitative research in family planning and reproductive health care. PMID- 15530226 TI - Clinical governance for better sexual health services. PMID- 15530227 TI - Reproductive issues and learning disability: different perspectives of professionals and parents. PMID- 15530228 TI - Sexual health services. PMID- 15530229 TI - Sexual health entering primary care: is prevention better than cure? PMID- 15530230 TI - Progestogens: the good, the bad and the ugly. PMID- 15530231 TI - The invisible man. Artificial insemination by donor and the legislation on donor anonymity: a review. AB - Until recently, sperm donors have remained anonymous under the 1990 Human Fertilisation and Embryology Act. There has been a distinct international trend towards allowing children access to identifying information about their donor. The UK has followed and, subject to parliamentary approval, children born by donations made after 1 April 2005 will be legally entitled to information revealing the identity of their donor. The lifelong well-being of the child created must be the prime consideration in any reform of the law. PMID- 15530233 TI - Condom use. PMID- 15530234 TI - Service standards: chaperones and record keeping. PMID- 15530236 TI - Emergency contraception: latest changes. PMID- 15530237 TI - Missing IUD fragment. PMID- 15530239 TI - Role of nurses. PMID- 15530240 TI - Evidence-based medicine and guidelines. PMID- 15530241 TI - HIV risk taking. PMID- 15530250 TI - Small-animal X-ray dose from micro-CT. AB - The use of micro-CT in small animals has increased in recent years. Although the radiation levels used for micro-CT are generally not lethal to the animal, they are high enough where changes in the immune response and other biological pathways may alter the experimental outcomes. Therefore, it is important to understand what the doses are for a specific imaging procedure. Monte Carlo simulation was used to evaluate the radiation dose to small animals (5-40 mm in diameter) as a result of X-ray exposure. Both monoenergetic (6-100 keV) and polyenergetic (15-100 kVp) X-ray sources were simulated under typical mouse imaging geometries. X-ray spectral measurements were performed on a mouse imaging X-ray system using a commercially available X-ray spectrometer, and spectra from high-energy systems were used as well. For a typical X-ray system with 1.0 mm of added Al at 40 kVp, the dose coefficients (dose to mouse per air kerma at isocenter) were 0.80, 0.63, 0.52, and 0.44 mGy/mGy for mouse diameters of 10, 20, 30, and 40 mm, respectively. A number of tables and figures are provided for dose estimation over a range of mouse imaging geometries. PMID- 15530249 TI - Radiation-force assisted targeting facilitates ultrasonic molecular imaging. AB - Ultrasonic molecular imaging employs contrast agents, such as microbubbles, nanoparticles, or liposomes, coated with ligands specific for receptors expressed on cells at sites of angiogenesis, inflammation, or thrombus. Concentration of these highly echogenic contrast agents at a target site enhances the ultrasound signal received from that site, promoting ultrasonic detection and analysis of disease states. In this article, we show that acoustic radiation force can be used to displace targeted contrast agents to a vessel wall, greatly increasing the number of agents binding to available surface receptors. We provide a theoretical evaluation of the magnitude of acoustic radiation force and show that it is possible to displace micron-sized agents physiologically relevant distances. Following this, we show in a series of experiments that acoustic radiation force can enhance the binding of targeted agents: The number of biotinylated microbubbles adherent to a synthetic vessel coated with avidin increases as much as 20-fold when acoustic radiation force is applied; the adhesion of contrast agents targeted to alpha(v)beta3 expressed on human umbilical vein endothelial cells increases 27-fold within a mimetic vessel when radiation force is applied; and finally, the image signal-to-noise ratio in a phantom vessel increases up to 25 dB using a combination of radiation force and a targeted contrast agent, over use of a targeted contrast agent alone. PMID- 15530251 TI - Imaging reactive oxygen species in arthritis. AB - Reactive oxygen species (ROS) have been shown to play a role in the pathogenesis of arthritides. Luminol was used as the primary reporter of ROS and photons resulting from the chemiluminescence reaction were detected using a super-cooled CCD photon counting system. Luminol was injected intravenously into groups of animals with different models of arthritis. Imaging signal correlated well with the severity of arthritis in focal and pan-arthritis as determined by histological measurement of ROS by formazan. Measurements were highly reproducible, sensitive, and repeatable. In vivo chemiluminescence imaging is expected to become a useful modality to elucidate the role of ROS in the pathogenesis of arthritides and in determining therapeutic efficacy of protective therapies. PMID- 15530252 TI - In vivo imaging of retinoic acid receptor activity using a sodium/iodide symporter and luciferase dual imaging reporter gene. AB - Retinoic acids are natural derivatives of vitamin A, and play important roles in modulating tumor cell growth by regulating differentiation, thus suggesting the potential use of these derivatives in cancer therapy and prevention. To visualize the intranuclear responses of functional retinoic acid receptors, we have developed a dual-imaging reporter gene system based on the use of sodium/iodide symporter (NIS) and luciferase in cancer cell lines. NIS and luciferase genes were linked with an internal ribosome entry site, and placed under the control of an artificial cis-acting retinoic acid responsive element (pRARE/NL). After retinoic acid treatment, I-125 uptake by pRARE/NL transfected cells was found to have increased by up to about five times that of nontreated cells. The bioluminescence intensity of pRARE/NL transfected cells showed dose-dependency. In vivo luciferase images showed higher intensity in retinoic acid treated SK RARE/NL tumors, and scintigraphic images of SK-RARE/NL tumors showed increased Tc 99m uptake after retinoic acid treatment. The NIS/luciferase imaging reporter system was sufficiently sensitive to allow the visualization of intranuclear retinoic acid receptor activity. This cis-enhancer imaging reporter system may be useful in vitro and in vivo for the evaluation of retinoic acid responses in such areas as cellular differentiation and chemoprevention. PMID- 15530253 TI - Systemic or regional chemotherapy for liver metastases from colorectal cancer: has the wheel stopped spinning? AB - The optimal management of patients with liver metastases from colorectal cancer is confounded by the large number of options available. In addition to ablative therapy (surgery, cryosurgery, radiofrequency ablation, etc.), chemotherapy plays a key role in the management of these patients. With more effective systemic therapy, some commentators have questioned the role of intra-arterial chemotherapy. However, a careful review of the literature,as well as ongoing research efforts, suggest that this approach remains an important option for patients and physicians to consider in appropriately selected patients for both first- and second-line therapy in advanced disease as well as for adjuvant therapy following resection of liver metastases. PMID- 15530254 TI - A clinician's guide to hereditary colon cancer. AB - Approximately 10% of patients diagnosed with colorectal cancer are at risk for a hereditary form of the disease. At-risk patients can be offered genetic counseling and testing to determine whether they carry a detectable mutation for such a syndrome. If so, this information provides the clinician with valuable data about the patient's risk for other cancers, and what further surveillance and risk reduction options should be incorporated into the management plan. Mutation identification within a family also makes it possible for other family members to learn if they are at risk for the same syndrome. There are many hereditary colorectal cancer syndromes, and the clinician must know what essential information should be elicited from a family history and which patients should be referred for genetic counseling and testing. PMID- 15530255 TI - Evidence-based approaches to overcoming barriers to clinical trial accrual. PMID- 15530256 TI - Does Gleason pattern 5 on prostate biopsy specimens accurately predict treatment outcomes in patients with prostate cancer? PMID- 15530257 TI - Therapy for unresectable hepatocellular carcinoma: time for XRT? PMID- 15530258 TI - An evaluation of barriers to accrual in the era of legislation requiring insurance coverage of cancer clinical trial costs in California. AB - PURPOSE: Clinical trials are essential to improve cancer therapy, but only 3% of newly diagnosed adult cancer patients enroll annually. We previously conducted a prospective analysis of factors affecting trial accrual at the UC Davis Cancer Center between 1997 and 2000. It was found that the accrual rate was 14% and that patients with private insurance were significantly less likely than patients with government insurance to enroll, suggesting that fear of insurance denial was a barrier. In 2002, a new California law (SB37) required insurers to reimburse routine costs of care for cancer trials. METHODS: To assess the impact of SB37 on accrual, we repeated our study using the same sur vey instrument. Oncologists seeing new patients at the UC Davis Cancer Center from August to November 2002 completed questionnaires that inquired about patient characteristics and eligibility, protocol availability, and patient willingness to participate. RESULTS: Physicians considered clinical trials for 55% (118/216) of patients, but trials were available for only 53% (62/118). Eligibility criteria were met by 82% (51/62). Of these, 69% (35/51) agreed to participate (vs 51% previously). No patient declined to participate because of insurance limitations (vs 8% previously). Furthermore, insurance type was no longer a significant factor in determining whether patients would enroll. This suggests that although the overall rate of accrual is only slightly increased after passage of SB37, patients may be more willing to enroll. Efforts to increase participation must include enhancing physician and patient awareness of SB37. PMID- 15530259 TI - The prognostic significance of Gleason pattern 5 in prostate cancer patients treated with Pd 103 plus beam radiation therapy. AB - BACKGROUND: There is little clinical information specifically regarding the clinical significance of Gleason pattern 5 in prostate biopsies. Accordingly, we have analyzed the effect of pattern 5 cancer on the prognosis of prostate cancer treated with Pd-103 brachytherapy. METHODS: Intermediate-risk patients with a Gleason score of 7 or higher and/or a prostate-specific antigen level of 10-20 ng/mL and whose biopsy slides were available for review were treated on a randomized trial. The regimens consisted of implantation with Pd 103 (90 vs 115 Gy [National Institute of Standards and Technology; NIST-1999]), combined with 44 Gy versus 20 Gy of supplemental beam radiation, respectively. Beam radiation was delivered with a four-field arrangement, designed to cover the prostate and seminal vesicles with a 2-cm margin (reduced to 1.0 cm posteriorly). Isotope implantation was per formed by standard techniques, using a modified peripheral loading pattern. All prostate biopsy specimens were reviewed for Gleason score by one investigator (L. T.). Along with assignment of a Gleason score based on established criteria, the presence of any pattern 5 cancer was separately noted and photographed for future review. Freedom from biochemical failure was defined as a serum prostate-specific antigen level < or = 0.5 ng/mL at last follow-up. Four of the 156 patients had insufficient PSA follow-up for inclusion, leaving 152 patients for cancer control analysis. RESULTS: Overall actuarial biochemical freedom from failure was 86% at 3 years, with 20 patients having experienced biochemical failure. Patients with or without Gleason pattern 5 cancer in their biopsy specimen had similar overall biochemical control. There was no obvious trend toward poorer overall biochemical cancer control in patients with pattern 5 cancer, regardless of whether the pretreatment prostate-specific antigen was less than or greater than 10 ng/mL. Of the 17 patients with biochemical failure, clinically evident bone metastases has developed in five. Three of these five patients who had a positive bone scan had pattern 5 cancer in their biopsy. CONCLUSIONS: Although the presence of pattern 5 disease may be a risk factor for early systemic failure, we are encouraged that high-dose, brachytherapy-based treatment seems to provide a high likelihood of biochemical cancer control, even in patients with the highest-grade cancer. PMID- 15530260 TI - A comparison of chemoembolization combination with and without radiotherapy for unresectable hepatocellular carcinoma. AB - PURPOSE: This study evaluated the effect of transcatheter arterial chem oembolization combined with external beam radiotherapy on the response rates and sur vival of patients with unresectable hepato-cellular carcinoma. Transcatheter arterial chemoembolization is frequently used for the treatment of this cancer, but complete or massive necrosis is seldom observed. Historically, radiotherapy for hepatocellular carcinoma has yielded poor long-term survival. Multimodality therapy has been initiated in an effort to improve survival statistics. PATIENTS AND METHODS: We retrospectively studied 203 patients with unresectable hepa tocellular carcinoma, who were free of tumor thrombus, lymph node involvement, or extrahepatic metastasis based on computed tomography scans of the chest and abdomen. Among the 203 patients who received transcatheter ar terial chemoembolization as initial therapy, 54 also received combination therapy with external beam radiotherapy. Tumor response rate, survival, and failure patterns were analyzed and compared between the two groups. RESULTS: Objective responses (complete and partial responses) on computed tomography study were obser ved in 31% and 76% of patients in the non-radiotherapy and radiotherapy groups, respectively. Overall survival rates in the patients in the radiotherapy group were 71.5%, 42.3%, and 24.0% at 1, 2, and 3 years, respectively, improved over the non-radiotherapy group rates of 59.6%, 26.5%, and 11.1% at 1, 2, and 3 years, respectively. Intrahepatic failure was lower in the radiotherapy group than in the non-radiotherapy group, but the difference was not significant. Side effects from radiotherapy were common, but rarely severe. CONCLUSIONS: This retrospective study suggests that the outcome of unresectable hepatocellular carcinoma can be influenced by radiation therapy, but a prospective randomized trial would be necessary to draw definitive conclusions. PMID- 15530261 TI - Does screening or surveillance for primary hepatocellular carcinoma with ultrasonography improve the prognosis of patients? AB - PURPOSE: The purpose of this paper is to evaluate the efficacy of ultrasonographic screening for primary hepatocellular carcinoma. PATIENTS AND METHODS: A total of 680 eligible cases were classified into three groups (surveillance, opportunistic, and symptomatic groups) according to their initial exposure. We used survival time, tumor morphology, and T staging as prognostic outcomes. The outcomes of screened/unscreened and sur veillance/nonsur veillance were compared with the use of the logistic regression model. RESULTS: The adjusted odds ratios for the screened group versus the unscreened group, with 1-, 2-, and 3-year survival time being used as outcomes, were 0.33 (95% confidence interval [CI], 0.21-0.52), 0.33 (95% CI, 0.21-0.53), and 0.37 (95% CI, 0.23 0.61), respectively. The adjusted odds ratios for surveillance versus nonsurveillance were 0.58 (95% CI, 0.35-0.97), 0.45 (95% CI, 0.27-0.74), and 0.44 (95% CI, 0.26-0.73). The odds ratios were even smaller when tumor morphology or T stage was taken as the main outcome. All these results were statistically significant. There were significant gradient relationships between prognostic outcomes and extent of screening history. CONCLUSION: The significant impact of ultrasonographic screening on mortality reduction was demonstrated. These findings strongly suggest that early detection of hepatocellular carcinoma by ultrasound may improve the prognosis of patients with hepatocellular carcinoma. PMID- 15530262 TI - Randomized phase II trial of concomitant chemoradiotherapy using weekly carboplatin or daily low-dose cisplatin for squamous cell carcinoma of the head and neck. AB - PURPOSE: This randomized, phase II study aimed to compare concomitant chemoradiotherapy using weekly carboplatin or daily low-dose cisplatin as a treatment for squamous cell carcinoma of the head and neck. PATIENTS AND METHODS: One hundred nineteen patients with moderate- to advanced-stage disease were eligible for the study. Fifty-three patients had stage II disease, 28 had stage III, and the remaining 38 had stage IV disease. Primary tumor sites included the larynx (N = 63), oropharynx (N = 30), hypopharynx (N = 23), and oral cavity (N = 3). Each patient received either a weekly carboplatin dose (100 mg/m(2)) in one arm or daily cisplatin (4 mg/m(2)) in the other arm for the initial 4 weeks of radiotherapy. The radiotherapy dose of 65 Gy was given in 26 fractions over 45 days, dependent on a good tumor response at 40 Gy. For ty-nine (81.7%) of 60 patients treated with carboplatin and 41 (69.5%) of 59 patients treated with cisplatin received the full dose of radiotherapy. Surgical-resection was optionally used for the remaining patients. RESULTS: The median follow-up time was 63 months. The local control rate at 5 years was 56.2% for the carboplatin treated arm and 35.5% for the cisplatin-treated arm, respectively. The 5-year overall survival rate did not significantly differ between treatments: 71.4% for carboplatin and 66.0% for cisplatin. Hematologic toxicity was more frequent in the carboplatin-treated arm. No difference was observed in surgical complications or in radiation-related adverse effects. DISCUSSION: These findings suggest that weekly carboplatin treatment is preferable to daily low-dose cisplatin. This could be because the total dose of cisplatin was too low to be effective. PMID- 15530263 TI - Molecular markers and long-term survivors of glioblastoma multiforme. PMID- 15530264 TI - Is dentistry a profession? Part 3. Future challenges. AB - In 2 earlier articles a definition of professionalism was developed, and several specific professional responsibilities were deduced. This third and final article in the series examines whether dentistry qualifies as a profession. On various levels, the professionalism of dentistry is found wanting. However, attaining the status of a profession is a work in progress, which means that each profession will always have some deficiencies. The author concludes that dentistry qualifies as a profession but that it is also exhibiting a trend toward once again becoming a business (as it was before the 19th century). For the sake of honesty with the public, dentistry must make a choice between these 2 models. PMID- 15530265 TI - Regional variation in dental procedures among people with an intellectual disability, Ontario, 1995-2001. AB - BACKGROUND: The literature indicates that people with an intellectual disability have a prevalence of dental caries that is either lower than or similar to that of the general population. However, many of their caries go untreated, and extractions are more often used as a means of treatment than in the general population. A substantial percentage (40%) of day admissions to hospital of people with intellectual disabilities in Ontario is related to dental diseases. In this paper, we examine whether rates of in-hospital dental procedures are evenly distributed across Ontario and discuss possible explanations for the findings. MATERIALS AND METHOD: A retrospective analysis was made of routinely collected hospital admission data for people with an intellectual disability. Age and gender-adjusted rates for dental procedures were calculated using the direct method of adjustment and 1996 census population estimates of Ontario. Three different summary measures for the assessment of regional variation were used. RESULTS: Two areas had dental procedure rates among those with an intellectual disability that were significantly lower than the overall Ontario rate: Hamilton Wentworth and Quinte-Kingston and Rideau. The 3 district health council areas with the highest rates for dental procedures were Niagara, Essex-Kent and Lambton, and Durham-Haliburton-Kawartha and Pine Ridge; all 3 rates were higher than the overall Ontario rate. CONCLUSIONS: The use of day surgery and in hospital visits to treat dental diseases in people with an intellectual disability varies considerably by region in Ontario. Observed differences may indicate inequities. PMID- 15530266 TI - Pigmented lesions of the oral cavity: review, differential diagnosis, and case presentations. AB - Pigmented lesions are commonly found in the mouth. Such lesions represent a variety of clinical entities, ranging from physiologic changes to manifestations of systemic illnesses and malignant neoplasms. Evaluation of a patient presenting with a pigmented lesion should include a full medical and dental history, extraoral and intraoral examinations and, in some cases, biopsy and laboratory investigations. In this paper, an algorithm is proposed for the assessment of pigmented lesions of the oral cavity, and 3 patients with such lesions are described. PMID- 15530267 TI - The Klippel-Feil syndrome: a case report. AB - Short neck and fusion of cervical vertebrae are observed in several genetic conditions and well-defined syndromes. An 8-year-old boy with a short neck, low set posterior hairline, deafness and limited neck motion was suspected of having such a condition. Clinical and radiographic examination led to the diagnosis of Klippel-Feil syndrome. PMID- 15530268 TI - Active nonsurgical decompression of large periapical lesions--3 case reports. AB - This paper describes a new nonsurgical approach for treating large periapical lesions which involves using a modified vacuum system within the root canal space. This new technique produces a vacuum effect in the periapical zone, which facilitates evacuation of large amounts of inflammatory fluids. This technique was used in 3 clinical cases, in which the patients presented with copious amounts of exudate draining from within the root canals. Clinical and radiographic results showed long-term resolution of the endodontic lesions. This treatment of the periapical pathology was consistent with the principles of conservative, nonsurgical endodontic procedures. PMID- 15530269 TI - Influence of natural fruit juices in removing the smear layer from root surfaces- an in vitro study. AB - Certain elements of a patient's diet may be associated with dentin hypersensitivity. The intent of this study was to evaluate the degree of removal of the smear layer from dentin surfaces by various fruit juices. A smear layer was created on extracted human teeth by manual scaling. The roots were reduced and distributed into 8 experimental groups. Distilled water was the negative control. The juices were applied by 2 methods: topical application and topical application with friction. Specimens were photomicrographed and graded according to an index of smear layer removal. With topical application, all but 2 of the tested substances resulted in significantly greater removal of the smear layer and opening of dentinal tubules than was the case with the negative control (p = 0.05); the exceptions were Gala apple and Italian grape juices, which were no different from the control. For the active application (with friction), most substances removed more smear layer than the control (p < 0.05); Gala apple, Italian grape and orange juices were similar to the control. For each of the tested substances, removal of the smear layer did not differ with the method of application (topical vs. friction; p > 0.05). It is concluded that natural fruit juices can remove the smear layer from dentin surfaces, and the efficacy of this removal varies with the type of juice. PMID- 15530270 TI - Hospital waste management. PMID- 15530271 TI - Comparison of 25-gauge, Quincke and Whitacre needles for postdural puncture headache in obstetric patients. AB - OBJECTIVE: To compare the frequency of postdural puncture headache (PDPH) and failure rate of spinal anesthesia using 25-gauge Quincke and 25-gauge Whitacre needles in obstetric patients. DESIGN: Single blinded, interventional experimental study. PLACE AND DURATION OF STUDY: This study was conducted at the Department of Anesthesiology, Pain Management and Surgical Intensive Care Unit, Dow University of Health Sciences and Civil Hospital, Karachi from November 1, 2003-April 15, 2004. PATIENTS AND METHODS: One hundred females, aged 18-35 years, ASA physical status I and II, with singleton pregnancy undergoing elective or emergency cesarean section under spinal anesthesia were randomly allocated to receive spinal anesthesia either by using 25-gauge Quincke or 25-gauge Whitacre needles. Patients were followed for 3 days postoperatively. Headache, its relation with posture, onset, duration, severity and response to the treatment were recorded. RESULTS: Compared with the Whitacre group, frequency of postdural puncture headache was significantly higher in Quincke group (*p=0.015), while the overall occurrence of non-postdural puncture headache (NPDPH) did not differ significantly between two groups (p=0.736). Most of PDPH developed on 2nd postoperative day, were mild in nature and resolved within 48 hours of their onset. There was no significant difference in the failure rate of spinal anesthesia in both groups (p=0.149). CONCLUSION: It is suggested that use of 25 gauge Whitacre needle reduces the frequency of PDPH without increasing the failure rate of spinal anesthesia in obstetric patients. PMID- 15530272 TI - Outcome of combined interferon-ribavirin in the treatment of chronic hepatitis C. AB - OBJECTIVE: To determine the outcome of 24 weeks combination treatment with Interferon and Ribavirin as initial therapy in chronic hepatitis C patients and frequency of associated adverse effects during the treatment. DESIGN: Descriptive study. PLACE AND DURATION OF STUDY: Medical Unit, DHQ Hospital, Daggar, District Buner, NWFP, Pakistan, from January 1999 to June 2002. PATIENTS AND METHODS: Three hundreds and fifty consecutive non-cirrhotic, chronic hepatitis C patients with positive HCV anti bodies by 3rd generation ELISA, positive HCV RNA by polymerase chain reaction (PCR) and raised serum alanine amino transferase (ALT) were evaluated and treated with combination of interferon alfa 2-b three million units subcutaneously three injection weekly and ribavirin 800-1200 mg orally daily for 24 weeks and followed for another 6 months. The HCV RNA was checked after 12 weeks and at the end of the treatment. The ALT level was checked after 04 weeks, at 12 weeks, and at the end of the treatment. End of the treatment viral response, sustained viral response and side effects of therapy were noted. RESULTS: Normal ALT was observed in 260 (74.2%) of the patients after one month of treatment and 310 (88.5%) of the patients at the end of 03 months. HCR RNA was not detectable in 275 (78.5%) of the patients at the end of 03 months (12 weeks of treatment). At the end of 24 weeks of treatment the HCV RNA was not detectable in 298 (85.14%) patients. Two hundred and seventy-six (78.85%) of the patients had persistently normal ALT and HCV RNA in the six months' follow up after treatment. CONCLUSION: Combination treatment of interferon and ribavirin for 24 weeks produced 88.57% biochemical and 85.14% virological response. The side effects of anti-viral therapy were tolerable and manageable. PMID- 15530273 TI - Red cell immunization in beta thalassaemia major. AB - OBJECTIVE: To find out the frequency, pattern and factors influencing red cell immunization secondary to multiple blood transfusions in patients of beta thalassaemia major. DESIGN: A cross-sectional study. PLACE AND DURATION OF STUDY: Armed Forces Institute of Transfusion, Rawalpindi, in November 2002. PATIENTS AND METHODS: One hundred and sixty-one patients suffering from beta-thalassaemia major and on regular blood transfusions were included in the study. Their blood samples were tested for blood grouping, direct antiglobulin test and antibody screening/identification using reagents of DiaMed-ID Gel microtyping system. RESULTS: The total rate of red cell immunization was found to be 6.84%. Red cell alloantibodies were detected in 4.97% patients, and belonged mainly to Rh system, with one example each of anti-K, anti-Jsb and anti-Jka. Direct antiglobulin test was positive in 3 patients (1.87%) with increased hemolysis. Two had warm panreactive IgG antibodies suggesting red cell autoimmunization. Red cells of the 3rd patient showed sensitization with c-3d, with presence of an autoreactive cold agglutinin in the serum having a titre of 1:4. The red cell alloantibody formation was not influenced by age at first transfusion, number of blood transfusions and ethnicity. CONCLUSION: The rate of red cell alloimmunization in beta-thalassaemia major is relatively low in our setup and may be related to red cell homogeneity between the donor and recipient population. Routine pre transfusion matching of blood, other than ABO and Rh "D" antigens is not recommended because of low rate of red cell alloimmunization, and high costs associated with such testing. Hyperhaemolysis, due to acquired red cell autoantibodies was found to be an important complication. Patients who develop this complication should be tested for presence of underlying alloantibodies and considered for immunosuppressive treatment. PMID- 15530274 TI - Staphylococcus aureus and MRSA nasal carriage in general population. AB - OBJECTIVE: Assessment of Staphylococcus aureus (S. aureus) and MRSA nasal carriage in our general population. DESIGN: Cross-sectional analytical study. PLACE AND DURATION OF STUDY: A private Pathology Laboratory in Lahore city from January 2002 to December 2003. MATERIALS AND METHODS: Nasal swabs were obtained from the apparently healthy persons accompanying the patients attending the laboratory. These swabs were inoculated on blood and mannitol salt agar plates and incubated at 37 degrees C. The isolate(s) were identified as S. aureus on the basis of colony morphology, Gram staining, catalase and coagulase tests. Sensitivity to Oxacillin was determined using standard Kirby Bauer technique. RESULTS: Nasal swabs from 1660 subjects from the community were studied. Out of these samples, a total of 246 (14.82%) samples were positive for growth of S. aureus (nasal carriers of S. aureus). Out of 246 S. aureus isolates, 48 (19.51%) isolates were methicillin resistant S. aureus (MRSA). Nasal carriage was higher in males (15.47%) as compared to females (13.26%), in urban areas (16.99%) as compared to rural areas (11.32%) and in the year 2002 (16.02%) as compared to year 2003 (13.08%). However, the difference was significant only for the urban vs. rural comparison (p<0.05). MRSA isolates were also significantly higher among S. aureus isolates from the nasal cavities of urban subjects (22.98%) as compared to rural ones (11.11%). Maximum nasal carriage was present in the age group upto 9 years (20.23%) with decrease in the age groups 10-19, 20-29 and 30-39 years followed by small rise in the older subjects. CONCLUSION: The nasal S. aureus carriage as well as methicillin resistance among these isolates are more common in urban community. PMID- 15530275 TI - Evaluation of nutritional status of patients on haemodialysis. AB - OBJECTIVE: To evaluate the nutritional status of patients on haemodialysis for chronic renal failure. DESIGN: Single center cross-sectional hospital based study. PLACE AND DURATION OF STUDY: This study was carried out at haemodialysis unit of Shaikh Zayed Hospital, Lahore from March to May 2000. PATIENTS AND METHODS: Fifty-one patients on regular haemodialysis were included in the study. Nutritional status was checked by laboratory data (hemoglobin, hematocrit, blood urea nitrogen, serum albumin, total protein, and lipid profile), anthropometric measurements (height, dry weight, body mass index (BMI), mid arm circumference (MAC), triceps skin fold thickness (TSF) and mid arm muscle circumference (MAMC)) and symptoms. RESULTS: The total number of patients was 51. Twenty-four patients were male and 27 female. Mean age was 43 years. Majority of the patients, 27 (53%), was between 21 to 50 years of age. Major cause of end-stage renal disease was chronic glomerulonephritis in 19 (35%) patients followed by diabetic nephropathy 14 (27%) and hypertensive nephropathy in 11 (21%). Hemoglobin and hematocrit was normal only in 7 patients and rest of the 44 patients were anemic. Serum albumin was 3.64+/-0.59 gm/dl (Mean +/- SD), total protein 7+/-0.86 gm/dl, serum calcium 9.5+/-0.7 mg/dl, serum phosphorus 4.8+/-0.5 mg/dl, serum cholesterol 170+/-43.56 mg/dl,LDL 96.94+/-40.76 mg/dl, HDL 45.7+/-19.61 gm/dl, serum triglyceride 176.8+/-99 gm/dl. Adequacy of dialysis, urea reduction ratio (URR) was adequate only in 16(31%) and rest of the patients 35(69%) were getting inadequate amount of dialysis. On anthropometric measurements, majority of the patients was malnourished and in moderate to severe category of malnutrition. Symptoms of uremia (nausea, vomiting, body aches, pain and pulmonary oedema) were more in patients who were getting inadequate dialysis i.e. urea reduction ratio more than 70% (p-value<0.05). CONCLUSION: Majority of patients was anaemic, undernourished and factors responsible for the same were economical, late referral to nephrologist, inappropriate dietary restrictions and inadequate dialysis. PMID- 15530276 TI - Microalbuminuria as an indicator of kidney function among diabetics. AB - OBJECTIVE: To find out the role of microalbuminuria as an indicator of kidney function among diabetics. DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Diabetes control clinic of Shaikh Zayed Hospital, Lahore, a tertiary care hospital, was the venue. The study was conducted during March 2002 to August 2003. PATIENTS AND METHODS: Timed overnight urine samples were collected from all subjects and albumin was estimated using Radioimmunoassay (RIA) method. Albumin excretion rate (AER) was assessed in three groups including normals, Type 1 and Type 2 diabetics. AER over 30 microg/min was considered as significant. RESULTS: A significantly higher proportion of AER was observed in both types of diabetes. A higher proportion of increased AER was observed in females of Type 2 diabetics. CONCLUSION: The study has indicated that irrespective of the duration and type of diabetes the damage to the kidney can be avoided if good glycemic control is achieved. PMID- 15530277 TI - Effect of fiber bread on the management of diabetes mellitus. AB - OBJECTIVE: To evaluate the effects of soluble fiber rich bread on lowering blood glucose, blood pressure and blood lipid levels. DESIGN: An interventional study. PLACE AND DURATION OF STUDY: The study was undertaken at Medical Unit III, Jinnah Postgraduate Medical Centre, Karachi. Duration of the project was one year (1997 1998). SUBJECTS AND METHODS: Twenty stable diabetics of both genders were included in this study. They were provided with this bread (test diet) to replace their usual breakfast bread and chapati for lunch and dinner. The intakes of other foods as well as medications were unchanged. Anthropometric measurements, fasting and 2 hours postprandial blood sugar, fasting serum cholesterol and triglyceride levels were recorded before and after 4 weeks of intervention. Blood sugar and blood pressure were checked every week. Quality of life was assessed subjectively with the help of questionnaire, which were asked before and after the end of the intervention. RESULTS: Glycemic control, both fasting and postprandial, improved significantly during intervention. Satisfactory reduction of blood pressure as well as serum cholesterol and triglyceride level was also observed in these cases. The medicines reduced significantly and quality of life improved in all subjects. None of these subjects complained any gastrointestinal discomfort during their consumption of this bread for 4 weeks. CONCLUSION: The use of soluble fiber rich bread helps to control blood sugar, the hyperlipidemia, and blood pressure which are elevated in poorly controlled diabetics. Thus, integrated improvement will improve the quality of life, reduce the cost of drugs as well as help to avoid harmful effect of drugs. PMID- 15530278 TI - Lupus tumidus: a variant of cutaneous lupus erythematosus. AB - Lupus tumidus is a rare sub-type of chronic cutaneous lupus erythematosus characterized by dermal plaques in which excessive mucin accumulates early in disease process. We report a middle aged women having succulent, edematous and persistent plaque over her face for five years that was not responding to various empirical treatments offered to her. Finally, on clinico-pathological basis, it was diagnosed as a case of tumid lupus erythematosus (TLE) and she responded satisfactorily to the treatment regimen including oral steroids, chloroquine and application of sun screen. PMID- 15530279 TI - Follicular carcinoma thyroid in Pendred syndrome. AB - Pendred syndrome is a rare inherited disorder of bilateral sensorineural deafness with goitre. Presence of follicular carcinoma thyroid makes it even rarer. A case of a young girl is described with the features of Pendred syndrome and euthyroid state. Follicular carcinoma of thyroid was detected on histopathology requiring re-tailoring of the management. PMID- 15530280 TI - Lingual thyroid as a cause of snoring and sleep apnea. AB - Lingual thyroid is a rare developmental abnormality. We describe a case of lingual thyroid in a 28 years old female who presented with symptoms of heroic snoring and obstructive sleep apnea. She was treated with surgery and was advised a life long thyroid hormone replacement therapy. PMID- 15530281 TI - Threaded-alive on a wooden picket. AB - A case of 4-year-old girl is presented, brought threaded-alive on wooden picket due to head-on fall from roof with wooden picket entering through left subcostal region and exiting from right buttock. Midline laparotomy was performed. Wooden picket was removed after enlarging the entry and exit wounds. PMID- 15530282 TI - Granulomatous hypophysitis mimicking pituitary adenoma. AB - Granulomatous inflammation of the pituitary gland is a rare disease that mimics non-functioning pituitary adenoma. Its incidence is high in middle aged and elderly women. Two such cases of histopathologically confirmed granulomatous hypophysitis are reported which presented with clinical and radiological impression of pituitary adenoma. PMID- 15530283 TI - Postfixation pseudoaneurysm of the superficial femoral artery in femoral shaft fracture. AB - This case report describes a young female child presenting with a painful, expansile swelling at the medial aspect of her right thigh which had followed an open reduction internal fixation of a closed fracture of mid-shaft of right femur. An angiogram confirmed pseudoaneurysm of the superficial femoral artery probably caused by overpenetration of the drill bit or any sharp instrument while applying dynamic compression plate. It was treated surgically by resection of the aneurysm, reconstruction with inter-positional sephanous vein graft and removal of the hard-ware. PMID- 15530284 TI - Down syndrome with morgagni hernia and hypothyroidism. AB - A rare association of Down syndrome with hypothyroidism and Morgagni type of diaphragmatic defect is described in a one-year-old male child who presented with a history of recurrent chest infections and delayed milestones. He showed obvious features of Down syndrome but was also found to have hypothyroidism on investigations. Morgagni hernia was suspected on lateral chest X- ray and confirmed by CT scan thorax. He responded well to the management, which included surgical repair of the hernia. PMID- 15530285 TI - Successful percutaneous coil occlusion of a large pulmonary arteriovenous malformation. AB - Pulmonary arteriovenous malformation is one of the rarest congenital anomalies of cardiovascular system. We present a case of 30-year-old female with a large pulmonary arteriovenous malformation (PAVM) arising from the right lower pulmonary artery and draining to the left atrium. She underwent successful embolization using three detachable Cook coils. PMID- 15530286 TI - Microbiology services in public sector hospitals of Pakistan. AB - The purpose of clinical microbiology laboratory is to identify the actual etiologic agent(s) of infectious disease(s) based on morphological, biochemical, immunological and molecular procedures. Proficiency in isolating and identifying microorganisms, determining drug resistance of isolates; and in reporting the results is essential for the early detection and treatment of nosocomial pathogens. A Microbiologist is essentially interlinked with committees regarding hospital policy-making, infection-control, surveillance, investigation of infections in patients and personnel and maintaining a continuing education program. In the final analysis, the patients' well-being and health benefit most from clinical microbiology laboratory. This overview emphasizes the role of microbiology laboratory in government hospitals of Pakistan and general health care, without which the problem of multiple antibiotic resistance can only become worse. PMID- 15530287 TI - Factor V Leiden in young patients with thrombophilia. PMID- 15530288 TI - Anxiety and depression among young adults with chest pain. PMID- 15530289 TI - Serological profile of incidentally detected asymptomatic HBsAg positive subjects. PMID- 15530290 TI - A need to change behavior and attitudes in medical practice. PMID- 15530291 TI - Vibration anesthesia: a noninvasive method of reducing discomfort prior to dermatologic procedures. AB - In some dermatologic and cosmetic procedures, local anesthesia is not sufficient for relieving pain; often patients are averse to injections. We propose vibration anesthesia, the use of vibration delivered with commercially available inexpensive massagers to reduce discomfort. We find the analgesic effect of vibration helpful in minimizing pain in patients undergoing injections of botulinum toxin type A treatment for hyperhidrosis, injection of filler substances such as Restylane and Juvederm, laser therapy for leg veins, nail-fold injections, Q-switched laser treatment of tattoos, incision and drainage of abscesses, and cautery of facial warts, as well as facilitating anesthetic injections for needle-phobic patients. We expect that additional uses will be found as experience with this modality grows. Although the use of vibration anesthesia generally does not eliminate pain completely, it can serve to make the injection or procedure much more tolerable. PMID- 15530292 TI - Treatment of HIV-associated facial lipoatrophy with Radiance FN (Radiesse). AB - HIV treatments frequently include the administration of Highly Active Antiretroviral Therapy (HAART) to combat the disease; although no conclusive evidence exists, it has been suggested that the HAART regimen may lead to facial lipoatrophy, exemplified by fat loss from facial areas, most notably the cheeks. Lipoatrophy can cause severe psychosocial consequences because it can be an identifying marker of HIV infection. Calcium hydroxylapatite (Radiance FN, recently renamed Radiesse) has been postulated to be a long-lasting, biocompatible filler that does not ossify, migratem or calcify. We present three patients with HIV-associated lipoatrophy who were treated with Radiance FN. Touch up treatments were performed 1 month later for two patients. The percentage of improvement following the initial treatment ranged from 75 to 90 percent. Although there has been some loss of improvement over time, there has been significant persistence for up to 9 months. No complications have occurred, and patients were highly satisfied with treatment results. Our experience with Radiance FN has shown it to be a potentially valuable tool for treating facial lipoatrophy. PMID- 15530293 TI - Continuous-wave Nd:Yag laser hyperthermia: a successful modality in treatment of basal cell carcinoma. AB - It was observed that malignant tumor cells are more sensitive than normal cells to heat. Hyperthermia is known to be cytotoxic at temperatures above 41 degrees C and selectively lethal to cancer cells. The aim of the present study was to evaluate the therapeutic efficacy of continuous wave Nd:Yag laser-induced hyperthermia in treatment of basal cell carcinoma (BCC). The study was performed between April 1995 and August 2000 on 37 patients with BCC selected from of the outpatients of the dermatology clinic of al-Minya University Hospital. Patients were treated with continuous-wave Nd:Yag laser hyperthermia at 6-week intervals (laser output power was 10 W, spot size 8 mm, and irradiation time up to 1 minute). Following this treatment, 36 patients (97.3 %) were completely cured. Within a follow-up period of 3-5 years only one recurrence was encountered (2.7 %). Continuous wave Nd:Yag laser-induced hyperthermia should be considered as an alternative treatment for BCC. This simple, bloodless, outpatient procedure showed excellent efficacy and cosmetic result with minimal complication. PMID- 15530294 TI - Treatment of psoriasis with light and heat energy (LHE): a preliminary study. AB - In psoriasis the earliest observable electron microscopic changes are in the papillary dermis vasculature. Selective photothermolysis of the dermal vasculature using a 585-nm pulsed-dye laser (PDL) for the treatment of psoriasis was reported more than a decade ago. Although clinically effective, the pulsed dye laser has limited practicality because of small spot size (5-10 mm) and lack of cost effectiveness. The objective of this study is to evaluate the safety and efficacy of a novel Light and Heat Energy (LHE) selective-photothermolysis system for the treatment of psoriasis. This system delivers controlled heat simultaneously with the light pulse; it has a large beam spot (55 x 22 mm) and is sufficiently cost effective to be widely used. Thirteen patients, seven with plaque psoriasis, five with palmoplantar psoriasis, and one with guttate psoriasis participated in a controlled study. Patients received two therapy regimens in parallel: LHE plus salicylic acid or salicylic-acid cream alone as control. Treatments were applied twice weekly spread evenly over 4-6 weeks. In most cases a fluence of 8.6 J/cm2 was applied. Efficacy of the treatment was assessed using the psoriasis-severity index (PSI) calculated from the investigator's overall assessment of the plaque's erythema, scaling, and induration. PSI was evaluated at baseline, at every other treatment visit, and during two follow-up visits scheduled 1 and 5 weeks following the last treatment. In eleven patients who completed the treatment regimen, the average global PSI score for the treated sites was reduced by 65 percent. In contrast, the control sites average global PSI score was reduced by only 7.4 percent. Six patients (55 %) had a significant improvement of 70-percent reduction or above in their PSI score, ten patients (91 %) had a 50 percent or above reduction; only one patient had a poor response (12.5 % reduction in global PSI). Erythema was reported in all cases and resolved within 24-48 hours. Six cases of burns were resolved within 2 days to 2 weeks following appropriate remedy. No cases of dyspigmentation were encountered. Light and Heat Energy (LHE) selective photothermolysis combined with salicylic acid appears to be a practical modality for the treatment of mild to moderate psoriasis. Patients experience discomfort that is only mild. There is no convalescence downtime. Clinical results obtained are similar to those reported with the pulsed-dye laser; however, this new system seems more suitable to be widely adopted as a treatment modality. Further followup is required to determine whether remission periods will be as long as those reported for treatment with the pulsed-dye laser. PMID- 15530295 TI - Pigmented spindle-cell nevus: a melanoma simulator. PMID- 15530296 TI - Childhood leishmaniasis: report of 106 cases. AB - In Tunisia there are three epidemic clinical forms of cutaneous leishmaniasis. They are associated with three different species of Leishmania and are observed in different geographical areas. We undertook a single-center retrospective analysis of childhood leishmaniasis in order to describe epidemio-clinical profile, therapeutic characteristics and clinical outcomes of affected patients. The study comprises 166 children with 132 lesions of cutaneous leishmaniasis. The subjects ages range from 5 months to 15 years (average 8.75 years). The F:M sex ratio is 1.3. Leishmaniasis affects grown-up children in 74.5 percent of the cases. All of our patients live in an endemic area. The face is affected in 76.5 percent of cases. Mucosal leishmaniasis is present in 9 children (6.8 %). Clinical diagnosis confirmed by the parasitologic smear or histopathological examination in 89.6 percent of the cases. Treatment with intralesional meglumine antimoniate is done for 67 patients; the treatment regimen is one local injection (1 ml/cm(2)) per week until recovery. Systemic meglumine antimoniate is the initial therapy for 25 patients. Meglumine antimoniate treatment is well tolerated with no side-effects. All leishmaniasis lesions heal within an average period of 2.18 months. Childhood cutaneous leishmaniais is common in Tunisia. It has the characteristics of sporadic leishmaniasis. Mucosal leishmaniasis has a favorable outcome with no destruction, nor scaring deformity. The standard treatment remains intralesional meglumine antimoniate. PMID- 15530297 TI - A Simplified Psoriasis Area Severity Index (SPASI) for rating psoriasis severity in clinic patients. AB - INTRODUCTION: The Psoriasis Area and Severity Index (PASI) is the most widely used tool to assess psoriasis disease severity in clinical trials, although it can be exceedingly cumbersome for use in daily clinical practice. Because clinical trials rely on the PASI for inclusion criteria, having a PASI score on a clinic patient may be useful for determining if the patient has a level of disease severity similar to that of patients treated in clinical trials. PURPOSE: The purpose of this study is to assess a simplified measure of psoriasis disease severity that is more conducive to use in general dermatology practice, the simplified PASI (SPASI). METHODS: We evaluate an area-weighted assessment of lesion severity composed of the sum of the average redness, thickness, and scaliness of all the psoriasis lesions multiplied by an estimate of total body surface area involved. The SPASI is mathematically derived from the PASI. The SPASI and PASI are not identical because of the categorical nature of area estimates used in the PASI. We use existing psoriasis-population data regarding the anatomical distribution of psoriasis lesions to create a simulated patient database. Monte Carlo analysis is then performed to determine the relation between the PASI and SPASI. RESULTS: For a sample population with a mean PASI score of 12.8, the mean SPASI was 14.2. Correlation between the PASI and the SPASI was high (r = 0.90). Bland-Altman analysis showed no consistent bias between the PASI and the SPASI. When attempting to identify simulated patients with a PASI score of 12 (an inclusion criterion for many clinical trials for severe psoriasis), SPASI was 97 percent sensitive and 66 percent specific. DISCUSSION: The SPASI is much less onerous than the PASI, requiring estimation of only four rather than sixteen independent variables. It provides a quick and practical estimate of disease severity similar to the PASI and can be used to communicate that patients have a level of disease severity similar or dissimilar to that of patients studied in clinical trials. PMID- 15530298 TI - Spontaneous acroangiodermatitis in a young woman. AB - We present a case of acroangiodermatitis that for nearly 15 years was unrecognized and treated with skin grafting and radiotherapy. This case is also unusual in that neither venous insufficiency nor an underlying arteriovenous malformation could be demonstrated. Acroangiodermatitis is infrequently reported; a review of the literature emphasizes the similarities to Kaposi sarcoma both clinically and histologically. Because treatment for Kapsoi's sarcoma and hemangioendothelioma involves skin grafting and radiotherapy, awareness of this entity by dermatologists, surgeons, and pathologists is important. PMID- 15530299 TI - Adenolipoma of the skin. AB - Adenolipoma of the skin is an unusual variant of lipoma recently described by Hitchcock et al. and characterized by the presence of normal eccrine sweat glands within a lipoma. We report a case and review the literature. A 45-year-old woman presented with a slow-growing, painless nodule on the thigh, clinically considered to be lipoma. Microscopically it comprised an adipose-tissue proliferation with a single eccrine secretory coil and associated duct in the periphery and in the center of the nodule. This benign lesion has been termed adenolipoma because of the presence of adipose tissue and eccrine glands. It probably represents only a histological curiosity in which the eccrine glands are entrapped by the adipose proliferation. Adenolipoma of the skin is a distinct lesion that can occur in the dermis or subcutaneous tissue. PMID- 15530300 TI - Congenital lower lip pits: a case report. AB - The Van der Woude syndrome is a rare autosomal-dominant condition typically comprising cleft lip or cleft palate and distinctive pits of the lower lips. The degree to which individuals carrying the gene are affected varies widely, even within families. The variable manifestations include lip pits alone, absent teeth, and isolated cleft lip and palate of varying degrees of severity. Other associated anomalies have also been described. We report a case of Van der Woud syndrome manifesting with isolated lower lip pits in an adolescent female with similarly affected family members. PMID- 15530301 TI - Multiple cutaneous reticulohistiocytomas in a patient with rheumatoid arthritis. AB - A 64-year-old woman with a long-standing peripheral symmetric polyarthritis with positive rheumatoid serology was evaluated for multiple asymptotic papulonodules of fingers, mentum, lower lip, ears, and eyelids. Histopathologic examination showed a dermal infiltrate composed of histiocytes, multinucleate giant cells with ground-glass cytoplasm, and lymphocytes, suggestive of reticulohistiocytoma. The possibilities of multicentric reticulohistiocytosis with positive rheumatoid serology or coexistence of multiple cutaneous reticulohistiocytomas and rheumatoid arthritis are discussed. PMID- 15530302 TI - Subcutaneous fat necrosis: report of two cases. AB - Subcutaneous fat necrosis of the newborn is an uncommon, benign process in full term infants or postmature neonates who experienced a perinatal distress. It is a transient condition of unknown origin; however, hypercalcemia may be a potentially life-threatening complication of this otherwise self-healing process. We report two cases of subcutaneous fat necrosis and discuss the clinical features, etiology, and complications of the disease. PMID- 15530303 TI - Alopecia areata of eyelashes: a subset of alopecia areata. PMID- 15530304 TI - Circumscribed scalp hair loss following multiple hair-cutter ant invasion. AB - A 32-year-old woman presented with an abrupt, localized loss of scalp hair that occurred on the previous day. Her nails, skin, and mucosae were normal. On the vertex of the scalp, there was a circular patch of alopecia; the hairs were broken at approximately equal lengths above the skin surface. Several erythematous macules were seen in the affected area, but scale, twisted hair, and exclamation-mark hairs were not present. Further examination revealed the presence of ants on the scalp. This patient is one of several referred to our department presenting with hair loss associated with hair-cutter ant invasion. PMID- 15530305 TI - Capillaritis as a potential harbinger of cutaneous T-cell lymphoma. PMID- 15530306 TI - Papulonodular mucinosis in systemic lupus erythematosus. AB - A 41-year-old man with systemic and serological manifestations of systemic lupus erythematosus presented with a diffuse eruption comprising annular plaques. Histopathology revealed diffuse deposition of mucin throughout the dermis, consistent with papulonodular mucinosis. This uncommon entity of unclear pathogenesis has been described in systemic lupus erythematosus, discoid lupus erythematosus, and subacute cutaneous lupus erythematosus. PMID- 15530307 TI - Partial lipodystrophy in a girl. AB - A 9-year old girl presented with a 2-year history of severe loss of subcutaneous fat in her face. Her medical history included a self-limited episode of jaundice and mild fever 2 years prior. Laboratory examination revealed low serum C3 but normal serum C4. Renal function tests were normal. Low C3 can be a warning sign in patients partial lipodystrophy; it may be a harbinger of renal insufficiency. Patients with partial lipodystrophy may be predisposed to renal failure, especially after pregnancy or the use oral contraceptive pills. PMID- 15530308 TI - Sclerodermatomyositis associated with severe arthritis. AB - Overlap syndromes are a spectrum of diseases with clinical features of two or more classic connective tissue diseases. Polymyositis or dermatomyositis that is associated with features of systemic sclerosis, also called sclerodermatomyositis, is an uncommon overlap syndrome, with a rare antibody directed against the nucleolar antigen PM-Scl. This disease generally has good prognosis. We report a case of sclerodermatomyositis with an aggressive pulmonary and articular course. PMID- 15530309 TI - Hereditary syringomas: a case report. AB - Syringoma is a benign neoplasm of eccrine origin. Clinically, it is an eruption of small translucent-to-yellowish papules. These lesions are firm, smooth, and approximately 1-3 mm in diameter. They are most commonly found around the eyes and on the upper cheeks of middle-aged women. Lesions sometimes develop on the abdomen, axillae, penis, vulva, and scalp. Involvement of the scalp may be indistinguishable from nonscarring alopecia. Familial cases have been reported, and there is an increased incidence of syringoma in adults with Down syndrome. Eruptive syringoma, a separate entity, presents mostly in adolescents as clusters of numerous papules on the upper half of the body. PMID- 15530310 TI - Facial cellulitis associated with Pseudomonas aeruginosa complicating ophthalmic herpes zoster. AB - Cellulitis is a rare and severe soft-tissue infection characterized by acute, diffuse, spreading inflammation, often associated with systemic symptoms such as malaise and fever. Surgery of the head and neck, dental infections, sinusitis, upper respiratory tract infections, and trauma are the most common portal of entry for pathogens in facial cellulitis. A very unusual case complicating an ophthalmic herpes zoster in a 74-year-old woman was observed at the department of dermatology, Cagliari University (Italy). Culture of skin swabs showed growth of numerous Gram-negative bacilli, further identified as Pseudomonas aeruginosa. Therapy with intravenous ciprofloxacin was promptly instituted on the basis of the culture and sensitivity report. She was initially treated with daily drainage and twice-daily topical fusidic acid. The lesion completely resolved in 4 weeks, and no general complications or recurrence have been observed for 6 months. Early recognition and management of facial cellulitis is mandatory to avoid serious and generalized complications. Pseudomonas aeruginosa is rarely reported in facial cellulitis; there are apparently no reports of this infection occurring as a complication of ophthalmic herpes zoster. Herpetic damage of the anatomic barrier as well as impairment of defense mechanisms because of decompensated diabetes mellitus may have facilitated the colonization and proliferation of this opportunistic pathogen in our patient. PMID- 15530311 TI - Ulcerated disseminated cutaneous leishmaniasis associated with vitiligo, hypothyroidism, and diabetes mellitus in a patient with Down syndrome. AB - We report a 35-year-old man who was referred to our dermatology department with multiple, nodular, ulcerated, and crusted lesions disseminated on the face, trunk, and extremities. He has a known diagnosis of Down syndrome. The past medical history also included vitiligo (for 20 years), hypothyroidism (for 2 years), and type-II diabetes mellitus (for 3 months). Direct smear of an ulcer was positive for leishmania. Skin biopsy confirmed the diagnosis. A leishmanin skin test was negative. Polymerase chain reaction (PCR) from two separate skin biopsies demonstrated the presence of Leishmania major. To our knowledge, this is the first report of disseminated cutaneous leishmaniasis (DCL) caused by L. major in Iran, and also the first report of association between DCL with Down syndrome, vitiligo, hypothyroidism, and diabetes mellitus. PMID- 15530312 TI - Audiovisual presentations in peer-reviewed medical literature. PMID- 15530313 TI - Successful treatment of granuloma faciale with tacrolimus. PMID- 15530314 TI - Intradermal drug delivery by low-frequency sonophoresis (25 kHz). AB - Ultrasound waves (US) have been proposed to facilitate the absorption of active compounds (transdermal delivery) stimulating some disaggregation of the horny layer and promoting convective movements within the epidermis. Drugs used for alopecia areata, melasma, and lentigo, although proved effective, have limited effects resulting from only partial penetration into the skin. This study has evaluated the efficacy of low-frequency sonophoresis (LFS) at 25 kHz produced by a sonicator apparatus for treatment of alopecia areata, melasma, and solar lentigo. Thirty patients affected by alopecia areata were treated by application of methylprednisolone or cyclosporine solution followed by LFS. In a case-control study 48 women with melasma and 48 with solar lentigo were also treated by depigmenting emulsion and LFS application. For alopecia areata after 36 applications with LFS and 3-month treatment, the results were 57 percent partial regrowth and 29 percent total with methylprednisolone, and 33 percent partial regrowth and 34 percent total when cyclosporine was used. For melasma and solar lentigines the results when the drug application was followed by LFS, were (after twice-a-week application for 3 months) 75 percent complete depigmentation and 25 percent partial for melasma, 43 percent total regression and 57 percent partial for solar lentigo. CONCLUSION: This is the first report of sonophoresis at a frequency of 25 kHz in dermatocosmetology. The study shows that LFS, a not aggressive technique, enhance penetration of topic agents obtaining effects at the level of the epidermis, dermis and appendages (intradermal delivery), giving better results in the treatment of some cosmetic skin disorders. PMID- 15530315 TI - [Clinical trials in children. New Royal Decree, old concepts]. PMID- 15530316 TI - [Ibero-American Society of Neonatology. Collaborative group for the improvement of clinical practice and research in neonatology]. PMID- 15530317 TI - [Malignant disease presenting as rheumatic manifestations]. AB - BACKGROUND: Musculoskeletal pain is a frequent complaint in pediatrics in both tertiary and primary care. Although musculoskeletal symptoms are not usually related to severe disease, they can represent the first manifestation of an occult malignancy. OBJECTIVE: To describe the clinical manifestations and complementary findings of the patients referred to a pediatric rheumatology unit with a final diagnosis of malignancy. MATERIAL AND METHODS: We performed a retrospective review of all the children referred to a pediatric rheumatology unit between 1992 and 2002 whose final diagnosis was cancer. RESULTS: Of 3,982 patients referred, 10 had a final diagnosis of neoplasia (0.25 %). Six children were diagnosed with acute lymphoblastic leukemia, two with Langerhans' cell histiocytosis, one with Ewing's sarcoma, and one with metastases from retinoblastoma. The most frequent symptoms were arthralgias, limp, or back pain. Laboratory studies revealed anemia and a significant increase in sedimentation rate and lactate dehydrogenase values. Imaging studies contributed significantly to the diagnosis. The diagnosis was confirmed by bone marrow aspirate in children with leukemia and by bone biopsy in those with bone tumors. CONCLUSIONS: Malignancies should be considered in the differential diagnosis of children with musculoskeletal pain, especially in the presence of organomegalies or abnormal laboratory or imaging studies. PMID- 15530318 TI - [Erythropoietin treatment in critically ill children]. AB - OBJECTIVE: To analyze whether erythropoietin treatment increases hemoglobin and decreases transfusion requirements in critically ill children. PATIENTS AND METHODS: We performed an observational, prospective study of 23 critically ill children aged between 1 month and 6 years. Recombinant human eritropoietin (rHuEPO) was administered at a dosage of 150-750 U/kg/week over 3 days. Hemogram, reticulocyte, iron metabolism, serum ferritin and transferrin were measured before treatment started and weekly thereafter. RESULTS: After erythropoietin treatment, hematocrit, hemoglobin and red blood cells progressively increased, with a maximal response in the sixth week. At the end of treatment, hemoglobin increased 1.68 g/dl, hematocrit by 5 % and erythrocytes 600,000/ml/mm3. Transfusion requirements decreased from 59 transfusions at baseline to 12 in the first week of treatment and none from the sixth week. No treatment-related adverse effects were observed. CONCLUSION: Erythropoietin can be an effective treatment for anemia in some critically ill children, decreasing the number of transfusions and increasing hemoglobin. PMID- 15530319 TI - [Erythema nodosum: etiological changes in the last two decades]. AB - OBJECTIVE: To study the etiological factors associated with erythema nodosum in children hospitalized between January 1985 and December 2003. MATERIAL AND METHODS: We performed a retrospective, descriptive study through review of the medical records of children with a diagnosis of erythema nodosum (MBDS-ICD 9-CM, code 6952). Forty-five patients (27 boys and 18 girls) aged between 10 months and 14 years were studied. The following variables were analyzed: age, sex, associated symptoms, complementary investigations (blood count, erythrocyte sedimentation rate, throat swab, chest x-ray, Mantoux test, ASLO titer, stool culture, serology) and final diagnosis. RESULTS: The most frequent etiology was tuberculosis (10 patients), followed by Salmonella enteritidis (7 patients), group A beta -hemolytic Streptococcus (3 patients), Salmonella typhimurium (2 patients), Campylobacter jejuni (2 patients), Yersinia enterocolitica (1 patient), infectious mononucleosis caused by Epstein Barr virus (1 patient), cat scratch disease (1 patient), BCG vaccination (1 patient), associated chronic hepatitis B infection (1 patient), and associated amoxicillin treatment (1 patient). Etiology was unknown in 15 patients. The last case of erythema nodosum associated with tuberculous infection dated back to 1991, after which the most frequent etiologic factors associated with erythema nodosum were gastrointestinal pathogens. CONCLUSIONS: According to our results, the main etiological factor currently associated with erythema nodosum is gastrointestinal infection. Consequently, stool cultures, especially when there are associated gastrointestinal symptoms, are essential. PMID- 15530320 TI - [Acute otitis media in a pediatric primary care unit]. AB - BACKGROUND: Acute otitis media (AOM) is a serious public health problem and is considered one of the most common diagnoses in pediatric outpatient units. The aim of this study was to asses the incidence and distribution of AOM by age, sex, number of visits per episode, and the most frequent symptoms and signs in four pediatric primary care units. PATIENTS AND METHODS: A 12-month (1 September 2000 to 31 August 2001), prospective study was carried out in a primary care center on the outskirts of Madrid. RESULTS: During the study period there were 1,098 consultations for AOM in 521 patients. The overall incidence per 100,000 children aged less than 14 years was 12,080 cases (95 % CI: 11,120-13,090). The highest incidence per 100,000 children was in children aged 12-23 months with 38,780 cases (95 % CI: 33,340-44,430). A total of 34.5 % of the children diagnosed with AOM were aged less than 2 years. Ninety-six percent of the children received antibiotics. Most (81.4 %) of the children were completely cured while symptoms persisted in 15.4 %. A minority of the children (3.5 %) were referred to the otolaryngology department. PMID- 15530321 TI - [Tap-water scald burns]. AB - OBJECTIVE: To describe the characteristics of patients with tap-water scalds admitted to our hospital and review current knowledge on their prevention. PATIENTS AND METHODS: We performed a retrospective study of admissions for tap water scalds. The medical records codified as water-tap scalds (MBDS; CIE-9-CM, 940.0-949.5, E.924.0, E.924.2) of patients admitted to our hospital for more than 24 hours from January 1 to December 31, 2003 were reviewed. The following variables were studied: age, sex, mechanism, extent and degree of burn, localization, length of hospital stay, treatment, requirement for skin grafting, mortality, and sequelae. RESULTS: Twenty-nine patients were treated in our hospital. Sixty-two percent were boys. Age ranged from 3 days to 9 years. Six patients (20.6 %) were aged less than 1 year, 19 (63 %) were aged between 1 and 3 years and four (13.8 %) were aged from 3 to 9 years. Body surface area was 10 % or less in 25 patients and more than 20 % in four. Five patients required skin autografting. In two patients, the scald was produced in the neonatal area of our hospital. The mean length of hospital stay was 12 days, ranging from 1 to 38 days. None of the patients died. One patient suffered severe sequelae. CONCLUSIONS: Because of the frequency and severity of the burns reported in this study, pediatricians should increase their efforts in educating families about this type of burn. In addition, effective legislation should be implemented in Spain. PMID- 15530322 TI - [Evaluation of renal stone disease: metabolic study]. AB - Renal stone formation is a multifactorial process in which all the information obtained from the patient (medical history, imaging tests, stone analysis, metabolic study and physicochemical urine analysis) shows a different facet of the same process. Consequently, all these investigations should be evaluated together. In half of all patients, stone formation is secondary to the presence of metabolic alterations in urine, of which the most frequent is idiopathic hypercalciuria. The second most frequent cause is infection and/or urinary malformations, while hereditary enzyme defects are highly unusual. Reference values for urinary excretion of lithogenic metabolites (calciuria, uricosuria, oxaluria, citraturia, etc.) are essential for an adequate metabolic study, since urinary excretion depends on multiple factors, which have been described in the various publications in the literature. Physicochemical study evaluating saturation of the various salts dissolved in urine should be performed. These saturations are currently considered to be a highly useful index for determining the risk of crystallization and stone formation in patients with lithiasis and for evaluating the effectiveness of treatment. Lastly, the metabolic profile of renal lithiasis in children resembles that in adults, suggesting that predisposition to renal lithiasis begins in childhood. The early detection of the metabolic alterations observed in these patients will reduce the incidence of this disease in both children and adults. PMID- 15530323 TI - [Codification of psychosocial diagnoses]. AB - Psychosocial pathology accounts for a significant proportion of pediatric practice. Its registry, however, is usually scanty or erratic. The purpose of this update was to collect and define psychosocial diagnoses within the ICM-9-CM coding system in order to aid pediatric practice and to offer interpretations of diagnoses, which would aid their coding. PMID- 15530325 TI - [Glycogen storage disease type IX presenting as abdominal distention, hepatomegaly and hypoglycemia during infancy]. AB - Glycogen storage diseases are a rare group of disorders in daily pediatric practice but must be taken into account when a patient presents with poor physical growth, hepatomegaly, hypoglycemia, hypotonia and/or other metabolic disturbances. Early diagnosis allows treatment that might improve the patient's outcome to be started or, at the very least, genetic counseling to be given to the parents. We present a 10-month-old boy who presented with growth retardation, abdominal distention and hepatomegaly and who was finally diagnosed with glycogenosis type IX. Definitive diagnosis was obtained by demonstrating the enzyme defect (phosphorylase beta-kinase) in affected tissues. Enteral nutrition was started using a diurnal high-carbohydrate diet with frequent feedings and nocturnal nasogastric continuous feeding, achieving optimal growth parameters and clinical response. PMID- 15530324 TI - [Hyperinsulinism-hyperammonemia syndrome due to a de novo mutation in exon 7 (G979A) of the glutamate dehydrogenase gene with excellent response to diazoxide]. AB - Hyperinsulinism-hyperammonemia syndrome is characterized by recurrent and symptomatic hypoglycemias in childhood, secondary to hyperinsulinism associated with mild and asymptomatic hyperammonemia. This syndrome is caused by dominantly expressed mutations of the glutamate dehydrogenase gene (10q23.3). These mutations modify control of enzyme activity and represent the second cause of congenital hyperinsulinism of known genetic etiology. Moreover, this syndrome is the first genetic disorder due to an increase of function in an enzyme of intermediary metabolism to have been identified. We present the case of a 16 month-old boy with symptomatic recurrent hypoglycemias from the end of the first year of life, caused by a de novo mutation in exon 7 (G979A) of the GDH gene, with excellent outcome after diazoxide treatment. PMID- 15530326 TI - [Spontaneous esophageal perforation in a preterm neonate]. PMID- 15530327 TI - [Do we respect children's rights?]. PMID- 15530328 TI - [Sodium and fluoride in drinking water in infancy]. PMID- 15530329 TI - [Criteria for the appropriateness of admissions in the pediatric emergency setting]. PMID- 15530330 TI - [Hypertrophic pyloric stenosis in twins]. PMID- 15530331 TI - [Pinealoma presenting as a psychiatric disorder]. PMID- 15530332 TI - [Autonomic dysfunction syndrome secondary to tuberculous meningitis]. PMID- 15530333 TI - [Cat scratch disease with hepatosplenic involvement]. PMID- 15530334 TI - [Newborn with bladder extrophy associated with finasteride exposure]. PMID- 15530335 TI - [Megacystis-microcolon-intestinal hypoperistalsis syndrome: apropos of a case of prolonged survival]. PMID- 15530336 TI - [Pigmentation band on toenail]. PMID- 15530337 TI - [Chronic obstructive pulmonary disease: an ambiguous name and an obstacle in the campaign against smoking]. PMID- 15530338 TI - [In support of the term chronic obstructive pulmonary disease (COPD)]. PMID- 15530339 TI - [Bronchoscopic lung biopsy with fluoroscopy to study 164 localized pulmonary lesions]. AB - OBJECTIVE: To assess the usefulness of bronchoscopic lung biopsy (BLB) in the diagnosis of localized pulmonary lesions. METHODS: A prospective, descriptive study of consecutive patients with a suspected neoplastic nodule, mass, or infiltrate with nonspecific bronchoscopic findings after performing BLB with fluoroscopy. The lesions, clinical and patient characteristics, and outcomes were evaluated. RESULTS: One hundred sixty-four patients with a mean (SD) age of 65 +/ 12 years and a total of 156 masses or nodules and 8 infiltrates (144 neoplasms and 20 nonneoplastic lesions) participated in the study. On average, the lesion was 4.3 +/- 2 cm in diameter and 6 +/- 2 BLBs were performed on each patient, with a rate of serious complications of less than 1%. BLB was diagnostic in 113 cases (69% of the neoplasms and 70% of the nonneoplastic). In cases diagnosed by BLB, the lesions were larger (4.6 +/- 2 cm), more samples were obtained (6 +/- 2 biopsies), nondiagnostic endoscopic alterations were found (30%), and the bronchus sign was present (22%). Moreover, bronchoscopy was better tolerated by those patients. CONCLUSIONS: BLB with fluoroscopy increased the diagnostic yield of bronchoscopy for localized lesions with nonspecific findings, regardless of etiology. Larger neoplasms, the bronchus sign, good tolerance, and more tissue samples increased the number of diagnoses, with few complications. PMID- 15530340 TI - [Predictors of outcome in blunt chest trauma]. AB - INTRODUCTION: Thoracic trauma is often associated with polytrauma. Because mortality is high, the search for prognostic tools is useful. PATIENTS AND METHODS: A total of 108 patients with blunt thoracic trauma, 73 of whom had multiple injuries, were studied in an intensive care unit (ICU). The variables named as potential predictors of outcome were the need for mechanical ventilation, duration of ventilation, and high positive end-expiratory pressure (PEEP); the presence of rib fractures, pulmonary contusion, pleural involvement (hemo- and/or pneumothorax), or lung infection; the need for emergency surgery; mean duration of ICU stay, and age. We also studied whether or not the mortality rate was higher in polytrauma patients. Student t and chi2 tests (95% confidence level) and multiple regression analysis (Hosmer-Lemeshow goodness of fit) were used to analyze the results. RESULTS: The need for mechanical ventilation, radiographic evidence of pulmonary contusion, emergency surgery, and hemodynamic instability were risk factors for increased mortality. Higher risk of mortality was not demonstrated for patients with multiple injuries. For patients in need of mechanical ventilation, high PEEP was a predictor of poor prognosis. CONCLUSIONS: The presence of the aforementioned predictors (mechanical ventilation, high PEEP, pulmonary contusion, emergency surgery, and hemodynamic instability) indicate serious injury to the lung parenchyma, which is the main determinant of outcome for patients with thoracic trauma. PMID- 15530341 TI - [Factors associated with smoking onset: 3-year cohort study of schoolchildren]. AB - OBJECTIVE: To analyze the predictors of smoking onset among schoolchildren. METHODS: A cohort study of 1056 children starting in first year secondary school at 44 schools in Barcelona was carried out. Participating children were invited to answer a lifestyle questionnaire every year for 4 years. Each questionnaire carried a personal code to allow the 4 questionnaires to be matched. Matching questionnaires were found for 729 children, 70% of the initial sample. RESULTS: Over the study period, the prevalence of regular smokers increased from 1.7% to 22% among boys and from 1.6% to 38.2% among girls. The predictors of smoking onset among boys were scoring high on the pro-smoking attitudes index (odds ratio [OR]=1.2; 95% confidence interval [CI], 1.1-1.3), intention to smoke in the future (OR=2.2; 95% CI, 1.0-4.9), low self-efficacy in resisting pressures to smoke (OR=0.98; 95% CI, 0.96-0.99), having siblings that smoke (OR=2.5; 95% CI, 1.2-5.4), and spending some free time in bars (OR=2.4; CI, 1.1-4.9). Among girls, the predictors were having low self-esteem (OR=0.94; 95% CI, 0.88-0.99), scoring low on the anti-tobacco attitudes index (OR=0.92; 95% CI, 0.88-0.97), having siblings who smoke (OR=2.5; 95% CI, 1.2-5.5), spending some free time in discotheques (OR=4.5; 95% CI, 1.9-11.8), and living in high socioeconomic-status neighborhoods (OR=3.1; 95% CI, 1.4-10.9). CONCLUSIONS: The results show the importance of cognitive variables as well as a variety of environmental variables, particularly the pattern of free time use and the influence of sibling models. Prevention programs must take into account smoking onset risk factors as a whole. PMID- 15530342 TI - [Risk factors of readmission in acute exacerbation of moderate-to-severe chronic obstructive pulmonary disease]. AB - OBJECTIVES: More effective management of chronic obstructive pulmonary disease (COPD) and improved survival of COPD patients requires a better understanding of the risk factors for exacerbation. The aim of this study was to identify factors related to readmission in patients with moderate-to-severe COPD. PATIENTS AND METHODS: Ninety patients with moderate-to-severe COPD hospitalized consecutively for acute exacerbation were studied prospectively. At discharge, the following potential predictors were assessed: clinical and spirometric variables, arterial blood gases, and respiratory muscle strength determined noninvasively. The patients were followed for a period of 3 months. Readmission for exacerbation and time intervals free of hospitalization were recorded. RESULTS: Univariate analysis showed that the presence of cor pulmonale (P<.05), long-term oxygen therapy (P<.05), hypercapnia (P<.05), and high inspiratory load--mean inspiratory airway pressure measured at the mouth exceeding 0.40 cm H2O and a pressure-time index greater than 0.25 (P<.05 for both variables)--increased the risk of hospitalization for exacerbation. Multivariate analysis showed that only cor pulmonale (P<.05) and a high pressure-time index (>0.25, P<.05) were independently related to risk of readmission. CONCLUSIONS: Cor pulmonale and a high pressure-time index are independent risk factors for hospitalization for exacerbation of moderate-to-severe COPD. PMID- 15530343 TI - [Leak monitoring in noninvasive ventilation]. AB - Nasal mask ventilation has been shown to be effective, but outcomes do not always match expectations because of mouth leaks, patient-ventilator asynchrony, or decreased upper airway patency. These developments are detected when they lead ultimately to circuit leaks that lower the effectiveness of ventilation through pressure loss, poor inspiratory triggering, and prolonged inspiratory time. The quality of sleep is affected, and adverse effects and treatment intolerance may arise. A number of ways to detect leaks and their practical consequences are proposed in this article. We applied 310 leak-detection procedures to 177 patients who had disappointing clinical, gasometric, or polysomnographic outcomes of ventilation. The leak-detection procedures varied according to the type of ventilation and the supposed underlying pathophysiological mechanism. Significant leaks were detected in 132 patients (76%); therapeutic changes were then prescribed to optimize outcomes. We present a practical method to apply in patients with suboptimal ventilation outcomes. If leaks can be detected during treatment, the probable cause of treatment failure can sometimes be established and possible pathophysiological mechanisms better understood. With this knowledge, it may be possible to improve ventilation. PMID- 15530344 TI - [Recommendations for the treatment of severe nosocomial pneumonia]. PMID- 15530345 TI - [Lung scintigraphy with technetium 99m depreotide in the assessment of solitary pulmonary nodules]. AB - A solitary pulmonary nodule (SPN) is defined as a parenchymal lesion measuring less than 3 cm in diameter that is not associated with other lesions. Ninety percent of SPNs are discovered incidentally and most are benign. The management of radiographically indeterminate SPNs has not been established and invasive procedures must be undertaken in order to understand the nature of the nodule. We review our experience with the use of somatostatin receptor scintigraphy with technetium Tc99m depreotide in 10 patients with suspected malignant SPN. We discuss the limitations and applications of this technique in the evaluation of whether SPNs are benign or malignant for the purpose of identifying patients for biopsy. For this application, this technique can be considered an alternative to positron emission tomography using fluorine-18 fluordeoxyglucose. PMID- 15530346 TI - [Lymphoid interstitial pneumonia resolved through antiretroviral therapy in an adult infected by human immunodeficiency virus]. AB - Lymphoid interstitial pneumonia (LIP) is a rare entity characterized by the infiltration of interstitial tissues and alveolar spaces by lymphocytes, plasma cells, and other lymphoreticular structures. The etiology of LIP is unknown, although associations with autoimmune and infectious factors have been described. The incidence of LIP has risen in recent years, mainly in children with acquired immunodeficiency syndrome (AIDS), while remaining less common in the adult population. No agreement has been reached regarding the diagnostic tests necessary for a firm diagnosis although suspicion is usually based on clinical and radiographic findings, with confirmation provided by examination of histological samples. The most common treatment is corticosteroids, either alone or in combination with other immunosuppressant agents although no evidence from controlled trials is available and cases have been reported in which LIP resolved in AIDS patients with antiretroviral therapy alone. We report the case of a human immunodeficiency virus-infected adult who was diagnosed with LIP by open lung biopsy and who responded to antiretroviral drugs with no need for associated corticosteroid therapy. PMID- 15530347 TI - [Cold abscess of the chest wall 58 years after thoracoplasty]. PMID- 15530348 TI - [Gold salt-induced lung disease]. PMID- 15530349 TI - Geometric morphometrics and paleoneurology: brain shape evolution in the genus Homo. AB - Paleoneurology concerns the study and analysis of fossil endocasts. Together with cranial capacity and discrete anatomical features, shape can be analysed to consider the spatial relationships between structures and to investigate the endocranial structural system. A sample of endocasts from fossil specimens of the genus Homo has been analysed using traditional metrics and 2D geometric morphometrics based on lateral projections of endocranial shape. The maximum and frontal widths show a size-related pattern of variation shared by all the taxa considered. Furthermore, as cranial capacity increases in the non-modern morphotypes there is a general endocranial vertical stretching (mainly centred at the anterior ascending circumvolution) with flattening and relative shortening of the parietal areas. This pattern could have involved some structural stress between brain development and vault bones at the parietal midsagittal profile in the heavy encephalised Neandertals. In contrast, modern humans show a species specific neomorphic hypertrophy of the parietal volumes, leading to a dorsal growth and ventral flexion (convolution) and consequent globularity of the whole structure. Brain tensors such as the falx cerebri have been hypothesised to represent one of the main physical constraints on morphogenetic trajectories, with additional influences from cranial base structures. The neurofunctional inferences discussed here stress the role of the parietal areas in the visuo spatial coordination and integration, which can be involved in higher cerebral functions and related to conceptual thinking. PMID- 15530350 TI - Discovery of a highly-specialized plesiadapiform primate in the early-middle Eocene of northwestern Africa. AB - In this paper we report the first occurrence of an endemic African plesiadapiform primate from the early-middle Eocene locality of Glib Zegdou (Hammada du Dra, Algeria). Dralestes (new genus) is a very specialized taxon, and its closest known relative is the enigmatic and controversial genus Azibius from Gour Lazib (Hammada du Dra). We group both together as the Azibiidae (new rank). Dralestes provides the first evidence of the upper dentition in this group. Some critical dental characters, such as a postprotocingulum on upper teeth, consistently reveal a primate status for the azibiids. Dralestes exhibits, however, a very unusual configuration of the upper molars by the enlarged parastyle, the lack of a metaconule, and the ectoloph structure (preparacrista, centrocrista and postmetacrista are aligned in a high blade-like structure). The apparent dental specializations of both lower premolars and molars of azibiids (exaenodonty, large P(4) bearing sharp apical cusps, and M(1) having a highly elongated trigonid) point to potential relationships with Chronolestes and carpolestid plesiadapiforms. A phylogenetic analysis, performed on 55 dental characters scored for 19 primate genera, clarifies the euprimate status of Altiatlasius, and thus indicates that azibiids are the only known plesiadapiforms from Africa. Azibiids are the sister group of the clade carpolestids/Chronolestes in the superfamily Plesiadapoidea. However, the azibiids differ fundamentally from carpolestids by the combined lack of a centroconule and multiple buccal cusps on P(4). The exact position of both Chronolestes and azibiids in the plesiadapoids appears difficult to resolve. A basal position of Chronolestes in this superfamily cannot be ruled out because it exhibits a simple morphology of I(1) and no conule on P(3). Considering this ad hoc hypothesis, azibiids are found to lie outside a clade including carpolestids/plesiadapids/saxonellids, and they are the sister group to Chronolestes. The clade including the carpolestid, saxonellid, and plesiadapid families is characterized by the occurrence of a centroconule on P(3-4). The lack of this trait in Dralestes and Chronolestes could mean that azibiids are basal plesiadapoids that diverged before the evolution of the common ancestor of the three derived plesiadapoid families, i.e. at least around the Paleocene-Eocene boundary or more probably during the Paleocene. The report of the first offshoot in Africa of plesiadapoids enhances the role of Africa in the early primate radiation. PMID- 15530351 TI - The Mio-Pliocene European primate fossil record: dynamics and habitat tracking. AB - We present here a study of European Neogene primate occurrences in the context of changing humidity. We studied the differences of primate localities versus non primate localities by using the mammal communities and the ecomorphological data of the taxa present in the communities. The distribution of primates is influenced by humidity changes during the whole Neogene, and the results suggest that the primates track the changes in humidity through time. The exception to this is the Superfamily Cercopithecoidea which shows a wider range of choices in habitats. All primate localities seem to differ from non-primate localities in that the mammal community structure is more closed habitat oriented, while in non primate localities the community structure changes towards open-habitat oriented in the late Neogene. The differences in primate and non-primate localities are stronger during the times of deep environmental change, when primates are found in their preferred habitats and non-primate localities have faunas better able to adapt to changing conditions. PMID- 15530352 TI - Disentangling Early Stone Age palimpsests: determining the functional independence of hominid- and carnivore-derived portions of archaeofaunas. AB - Determining the extent to which hominid- and carnivore-derived components of fossil bone palimpsests formed independently of each other can provide valuable information to paleoanthropologists interested in reconstructing the foraging adaptations of hominids. Because stone tool cutmarks, hammerstone percussion marks, and carnivore tooth marks are usually only imparted on bone during nutrient extraction from a carcass, these bone surface modifications are particularly amenable to the types of analyses that might meet this goal. This study compares the percentage of limb bone specimens that preserve evidence of both hominid- and carnivore-imparted bone damage from actualistic control samples and several Plio-Pleistocene archaeofaunas, including new data from Swartkrans Member 3 (South Africa). We argue that this procedure, which elucidates the degree of hominid-carnivore independence in assemblage formation, will allow researchers to extract for focused analyses high integrity components (hominid and carnivore) from presumably low integrity sites. Comparisons suggest that the hominid- and carnivore-derived components from sites in Olduvai Gorge Bed II (Tanzania), the ST Site Complex at Peninj (Tanzania), and Swartkrans Member 3 formed largely independent of each other, while data from the FLK 22 Zinjanthropus (FLK Zinj) site (Olduvai Gorge Bed I) indicate significant interdependence in assemblage formation. This contrast suggests that some Early Stone Age assemblages (e.g., the Olduvai Gorge Bed II sites, the Peninj ST Site Complex, and Swartkrans Member 3) are probably more useful than others (e.g., FLK Zinj) for assessing the maximal carcass-acquiring abilities of early hominids; in such assemblages as those in the former set, sole hominid-contribution is more confidently discerned and isolated for analysis than in assemblages such as FLK Zinj. PMID- 15530353 TI - Facial heights: evolutionary relevance of postnatal ontogeny for facial orientation and skull morphology in humans and chimpanzees. AB - Facial heights, i.e. the vertical distances between the superior and inferior limits of facial compartments, contribute to the orientation of the viscerocranium in the primate skull. In humans, vertical facial variation is among the main sources of diversity and frequently associated with an integrated suite of other cranio-mandibular traits. Facial heights and kyphosis are also important factors in interspecific variation and models of hominoid evolution. The ontogenetic determination of adult facial orientation and its relation to phylogenetic variation are unclear, but crucial in all previously mentioned respects. We addressed these issues in a sample of 175 humans and chimpanzees with Procrustes based geometric morphometrics, testing hypotheses of interspecific similarity in postnatal ontogenetic trajectories, early versus later ontogenetic facial pattern determination, and a developmental model of morphological integration. We analyzed the contribution of postnatal morphogenesis to adult vertical facial variation by partitioning morphological variation into a portion of pure growth allometry and a non-allometric fraction. A statistically significant difference of growth-allometries revealed that in both species growth established the adult skull proportions by vertical facial expansion, but while in chimpanzees the complete viscerocranium showed reorientation, in humans only the lower face was modified. In both species the results support a hypothesis of early facial pattern determination. A coincident emergence of morphological traits favors a hypothesis of developmental integration of the face, excluding traits of the basi- and neurocranium. Interspecific differences in integration may have implications for evolutionary studies. The present findings indicate that growth establishes the adult skull proportions and integrates principal facial orientation patterns, already there in early postnatal ontogeny. PMID- 15530354 TI - This little pIgR went to the mucosa. AB - In this issue, we report the structure of the terminal domain of the polymeric immunoglobulin receptor (pIgR), which mediates the "suicide" transcytosis of multimeric immunoglobulins (IgA, IgM). This assists in reconciling decades of biochemistry, revealing a long-puzzling interaction. PMID- 15530355 TI - Cytochrome p450 conformational diversity. AB - Cytochrome P450s display a remarkable range of conformations in parallel with activity toward a great diversity of substrates. This aspect of P450s now extends to include the dynamic behavior of the protein, as shown by recent crystal structures of Cyp51. PMID- 15530356 TI - First structural glimpse at a bacterial Ser/Thr protein phosphatase. AB - In this issue of Structure, we describe the crystal structure of the Ser/Thr protein phosphatase PstP from Mycobacterium tuberculosis, opening new perspectives to understand the putative roles of "eukaryotic-like" signaling elements in bacteria. PMID- 15530357 TI - Crystal structure of a polymeric immunoglobulin binding fragment of the human polymeric immunoglobulin receptor. AB - The polymeric immunoglobulin receptor (pIgR) is a type I transmembrane protein that delivers dimeric IgA (dIgA) and pentameric IgM to mucosal secretions. Here, we report the 1.9 A resolution X-ray crystal structure of the N-terminal domain of human pIgR, which binds dIgA in the absence of other pIgR domains with an equilibrium dissociation constant of 300 nM. The structure of pIgR domain 1 reveals a folding topology similar to immunoglobulin variable domains, but with differences in the counterparts of the complementarity determining regions (CDRs), including a helical turn in CDR1 and a CDR3 loop that points away from the other CDRs. The unusual CDR3 loop position prevents dimerization analogous to the pairing of antibody variable heavy and variable light domains. The pIgR domain 1 structure allows interpretation of previous mutagenesis results and structure-based comparisons between pIgR and other IgA receptors. PMID- 15530358 TI - Estriol bound and ligand-free structures of sterol 14alpha-demethylase. AB - Sterol 14alpha-demethylases (CYP51) are essential enzymes in sterol biosynthesis in eukaryotes and drug targets in antifungal therapy. Here, we report CYP51 structures in ligand-free and estriol bound forms. Using estriol as a probe, we determined orientation of the substrate in the active site, elucidated protein contacts with the invariant 3beta-hydroxy group of a sterol, and identified F78 as a key discriminator between 4alpha-methylated and 4alpha,beta-dimethylated substrates. Analysis of CYP51 dynamics revealed that the C helix undergoes helix coil transition upon binding and dissociation of a ligand. Loss of helical structure of the C helix in the ligand-free form results in an unprecedented opening of the substrate binding site. Upon binding of estriol, the BC loop loses contacts with molecular surface and tends to adopt a closed conformation. A mechanism for azole resistance in the yeast pathogen Candida albicans associated with mutations in the ERG11 gene encoding CYP51 is suggested based on CYP51 protein dynamics. PMID- 15530359 TI - An alternate conformation and a third metal in PstP/Ppp, the M. tuberculosis PP2C Family Ser/Thr protein phosphatase. AB - Serine/threonine protein phosphatases are central mediators of phosphorylation dependent signals in eukaryotes and a variety of pathogenic bacteria. Here, we report the crystal structure of the intracellular catalytic domain of Mycobacterium tuberculosis PstPpp, a membrane-anchored phosphatase in the PP2C family. Despite sharing the fold and two-metal center of human PP2Calpha, the PstPpp catalytic domain binds a third Mn(2+) in a site created by a large shift in a previously unrecognized flap subdomain adjacent to the active site. Mutations in this site selectively increased the Michaelis constant for Mn(2+) in the reaction of a noncognate, small-molecule substrate, p-nitrophenyl phosphate. The PstP/Ppp structure reveals core functional motifs that advance the framework for understanding the mechanisms of substrate recognition, catalysis, and regulation of PP2C phosphatases. PMID- 15530360 TI - The crystal structure of RhoA in complex with the DH/PH fragment of PDZRhoGEF, an activator of the Ca(2+) sensitization pathway in smooth muscle. AB - Calcium sensitization in smooth muscle is mediated by the RhoA GTPase, activated by hitherto unspecified nucleotide exchange factors (GEFs) acting downstream of Galphaq/Galpha(12/13) trimeric G proteins. Here, we show that at least one potential GEF, the PDZRhoGEF, is present in smooth muscle, and its isolated DH/PH fragment induces calcium sensitization in the absence of agonist-mediated signaling. In vitro, the fragment shows high selectivity for the RhoA GTPase. Full-length fragment is required for the nucleotide exchange, as the isolated DH domain enhances it only marginally. We crystallized the DH/PH fragment of PDZRhoGEF in complex with nonprenylated human RhoA and determined the structure at 2.5 A resolution. The refined molecular model reveals that the mutual disposition of the DH and PH domains is significantly different from other previously described complexes involving DH/PH tandems, and that the PH domain interacts with RhoA in a unique mode. The DH domain makes several specific interactions with RhoA residues not conserved among other Rho family members, suggesting the molecular basis for the observed specificity. PMID- 15530361 TI - Crystal structure of PriB, a component of the Escherichia coli replication restart primosome. AB - Maintenance of genome stability following DNA damage requires origin-independent reinitiation of DNA replication at repaired replication forks. In E. coli, PriA, PriB, PriC, and DnaT play critical roles in recognizing repaired replication forks and reloading the replisome onto the template to reinitiate DNA replication. Here, we report the 2.0 A resolution crystal structure of E. coli PriB, revealing a dimer that consists of a single structural domain formed by two oligonucleotide/oligosaccharide binding (OB) folds. Structural similarity of PriB to single-stranded DNA binding proteins reveals insights into its mechanisms of DNA binding. The structure further establishes a putative protein interaction surface that may contribute to the role of PriB in primosome assembly by facilitating interactions with PriA and DnaT. This is the first high-resolution structure of a protein involved in oriC-independent replisome loading and provides unique insight into mechanisms of replication restart in E. coli. PMID- 15530362 TI - Structural basis of CTP-dependent peptide bond formation in coenzyme A biosynthesis catalyzed by Escherichia coli PPC synthetase. AB - Phosphopantothenoylcysteine (PPC) synthetase forms a peptide bond between 4' phosphopantothenate and cysteine in coenzyme A biosynthesis. PPC synthetases fall into two classes: eukaryotic, ATP-dependent and eubacterial, CTP-dependent enzymes. We describe the first crystal structure of E. coli PPC synthetase as a prototype of bacterial, CTP-dependent PPC synthetases. Structures of the apo-form and the synthetase complexed with CTP, the activated acyl-intermediate, 4' phosphopantothenoyl-CMP, and with the reaction product CMP provide snapshots along the reaction pathway and detailed insight into substrate binding and the reaction mechanism of peptide bond formation. Binding of the phosphopantothenate moiety of the acyl-intermediate in a cleft at the C-terminal end of the central beta sheet of the dinucleotide binding fold is accomplished by an otherwise flexible flap. A second disordered loop may control access of cysteine to the active site. The conservation of functionalities involved in substrate binding and catalysis provides insight into similarities and differences of prokaryotic and eukaryotic PPC synthetases. PMID- 15530363 TI - Accurate detection of protein:ligand binding sites using molecular dynamics simulations. AB - Accurate prediction of location of cavities and surface grooves in proteins is important, as these are potential sites for ligand binding. Several currently available programs for cavity detection are unable to detect cavities near the surface or surface grooves. In the present study, an optimized molecular dynamics based procedure is described for detection and quantification of interior cavities as well as surface pockets. This is based on the observation that the mobility of water in such pockets is significantly lower than that of bulk water. The algorithm efficiently detects surface grooves that are sites of protein ligand and protein-protein interaction. The algorithm was also used to substantially improve the performance of an automated docking procedure for docking monomers of nonobligate protein-protein complexes. In addition, it was applied to predict key residues involved in the binding of the E. coli toxin CcdB with its inhibitor. Predictions were subsequently validated by mutagenesis experiments. PMID- 15530364 TI - Insights into DNA replication: the crystal structure of DNA polymerase B1 from the archaeon Sulfolobus solfataricus. AB - To minimize the large number of mispairs during genome duplication owing to the large amount of DNA to be synthesized, many replicative polymerases have accessory domains with complementary functions. We describe the crystal structure of replicative DNA polymerase B1 from the archaeon Sulfolobus solfataricus. Comparison between other known structures indicates that although the protein is folded into the typical N-terminal, editing 3'-5'exonuclease, and C-terminal right-handed polymerase domains, it is characterized by the unusual presence of two extra alpha helices in the N-terminal domain interacting with the fingers helices to form an extended fingers subdomain, a structural feature that can account for some functional features of the protein. We explore the structural basis of specific lesion recognition, the initial step in DNA repair, describing how the N-terminal subdomain pocket of archaeal DNA polymerases could allow specific recognition of deaminated bases such as uracil and hypoxanthine in addition to the typical DNA bases. PMID- 15530365 TI - Human leukotriene C(4) synthase at 4.5 A resolution in projection. AB - Leukotriene (LT) C(4) synthase, an 18 kDa integral membrane enzyme, conjugates LTA(4) with reduced glutathione to form LTC(4), the parent compound of all cysteinyl leukotrienes that play a crucial role in the pathobiology of bronchial asthma. We have calculated a projection map of recombinant human LTC(4) synthase at a resolution of 4.5 A by electron crystallography, which shows that the enzyme is a trimer. A map truncated at 7.5 A visualizes four transmembrane alpha helices per protein monomer. The densities in projection indicate that most of the alpha helices run nearly perpendicular to the plane of the membrane. At this resolution, LTC(4) synthase is strikingly similar to microsomal glutathione S transferase 1, which belongs to the same gene family but bears little sequence identity and no resemblance in substrate specificity to the LTC(4) synthase. These results provide new insight into the structure and function of membrane proteins involved in eicosanoid and glutathione metabolism. PMID- 15530366 TI - Structures of Selenomonas ruminantium phytase in complex with persulfated phytate: DSP phytase fold and mechanism for sequential substrate hydrolysis. AB - Various inositide phosphatases participate in the regulation of inositol polyphosphate signaling molecules. Plant phytases are phosphatases that hydrolyze phytate to less-phosphorylated myo-inositol derivatives and phosphate. The phytase from Selenomonas ruminantium shares no sequence homology with other microbial phytases. Its crystal structure revealed a phytase fold of the dual specificity phosphatase type. The active site is located near a conserved cysteine-containing (Cys241) P loop. We also solved two other crystal forms in which an inhibitor, myo-inositol hexasulfate, is cocrystallized with the enzyme. In the "standby" and the "inhibited" crystal forms, the inhibitor is bound, respectively, in a pocket slightly away from Cys241 and at the substrate binding site where the phosphate group to be hydrolyzed is held close to the -SH group of Cys241. Our structural and mutagenesis studies allow us to visualize the way in which the P loop-containing phytase attracts and hydrolyzes the substrate (phytate) sequentially. PMID- 15530367 TI - Crystal structures of Ral-GppNHp and Ral-GDP reveal two binding sites that are also present in Ras and Rap. AB - RalA is a GTPase with effectors such as Sec5 and Exo84 in the exocyst complex and RalBP1, a GAP for Rho proteins. We report the crystal structures of Ral-GppNHp and Ral-GDP. Disordered switch I and switch II, located away from crystal contacts, are observed in one of the molecules in the asymmetric unit of the Ral GppNHp structure. In the other molecule in the asymmetric unit, a second Mg(2+) ion is bound to the GppNHp gamma-phosphate in an environment in which switch I is pulled away from the nucleotide and switch II is found in a tight beta turn. Clustering of conserved residues on the surface of Ral-GppNHp identifies two putative sites for protein-protein interaction. One site is adjacent to switch I. The other is modulated by switch II and is obstructed in Ral-GDP. The Ral structures are discussed in the context of the published structures of the Ral/Sec5 complex, Ras, and Rap. PMID- 15530368 TI - Solution structure of the bacterial frataxin ortholog, CyaY: mapping the iron binding sites. AB - CyaY is the bacterial ortholog of frataxin, a small mitochondrial iron binding protein thought to be involved in iron sulphur cluster formation. Loss of frataxin function leads to the neurodegenerative disorder Friedreich's ataxia. We have solved the solution structure of CyaY and used the structural information to map iron binding onto the protein surface. Comparison of the behavior of wild type CyaY with that of a mutant indicates that specific binding with a defined stoichiometry does not require aggregation and that the main binding site, which hosts both Fe(2+) and Fe(3+), occupies a highly anionic surface of the molecule. This function is conserved across species since the corresponding region of human frataxin is also able to bind iron, albeit with weaker affinity. The presence of secondary binding sites on CyaY, but not on frataxin, hints at a possible polymerization mechanism. We suggest mutations that may provide further insights into the frataxin function. PMID- 15530369 TI - Structural insights into how the MIDAS ion stabilizes integrin binding to an RGD peptide under force. AB - Integrin alpha(V)beta(3) binds to extracellular matrix proteins through the tripeptide Arg-Gly-Asp (RGD), forming a shallow crevice rather than a deep binding pocket. A dynamic picture of how the RGD-alpha(V)beta(3) complex resists dissociation by mechanical force is derived here from steered molecular dynamic (SMD) simulations in which the major force peak correlates with the breaking of the contact between Asp(RGD) and the MIDAS ion. SMD predicts that the RGD alpha(V)beta(3) complex is stabilized from dissociation by a single water molecule tightly coordinated to the divalent MIDAS ion, thereby blocking access of free water molecules to the most critical force-bearing interaction. The MIDAS motif is common to many other proteins that contain the phylogenetically ancient von Willebrand A (vWA) domain. The functional role of single water molecules tightly coordinated to the MIDAS ion might reflect a general strategy for the stabilization of protein-protein adhesion against cell-derived forces through divalent cations. PMID- 15530370 TI - Large-scale structural changes accompany binding of lethal factor to anthrax protective antigen: a cryo-electron microscopic study. AB - Anthrax toxin (AT), secreted by Bacillus anthracis, is a three-protein cocktail of lethal factor (LF, 90 kDa), edema factor (EF, 89 kDa), and the protective antigen (PA, 83 kDa). Steps in anthrax toxicity involve (1) binding of ligand (EF/LF) to a heptamer of PA63 (PA63h) generated after N-terminal proteolytic cleavage of PA and, (2) following endocytosis of the complex, translocation of the ligand into the cytosol by an as yet unknown mechanism. The PA63h.LF complex was directly visualized from analysis of images of specimens suspended in vitrified buffer by cryo-electron microscopy, which revealed that the LF molecule, localized to the nonmembrane-interacting face of the oligomer, interacts with four successive PA63 monomers and partially unravels the heptamer, thereby widening the central lumen. The observed structural reorganization in PA63h likely facilitates the passage of the large 90 kDa LF molecule through the lumen en route to its eventual delivery across the membrane bilayer. PMID- 15530371 TI - The crystal structure of human CDK7 and its protein recognition properties. AB - CDK7, a member of the cyclin-dependent protein kinase family, regulates the activities of other CDKs through phosphorylation on their activation segment and hence contributes to control of the eukaryotic cell cycle. CDK7 also assists in the regulation of transcription as part of the transcription factor TFIIH complex. For maximum activity and stability, CDK7 requires phosphorylation, association with cyclin H, and association with a third protein, MAT1. We have determined the crystal structure of human CDK7 in complex with ATP at 3 A resolution. The kinase is in the inactive conformation, similar to that observed for inactive CDK2. The activation segment is phosphorylated at Thr170 and is in a defined conformation that differs from that in phospho-CDK2 and phospho CDK2/cyclin A. The functional properties of the enzyme against CDK2 and CTD as substrates are characterized through kinase assays. Experiments confirm that CDK7 is not a substrate for kinase-associated phosphatase. PMID- 15530372 TI - The X-ray structure of RANTES: heparin-derived disaccharides allows the rational design of chemokine inhibitors. AB - The biological activity of chemokines requires interactions with cell surface proteoglycans. We have determined the structure of the chemokine RANTES (regulated on activation normal T cell expressed) in the presence of heparin derived disaccharide analogs by X-ray crystallography. These structures confirm the essential role of the BBXB motif in the interaction between the chemokine and the disaccharide. Unexpected interactions were observed in the 30s loop and at the amino terminus. Mutant RANTES molecules were designed to abrogate these interactions and their biological activity examined in vivo. The K45E mutant within the BBXB motif lost the capacity to bind heparin and the ability to elicit cellular recruitment. The Y3A mutant maintained its capacity to bind heparin but was unable to elicit cellular recruitment. Finally, a tetrasaccharide is the smallest oligosaccharide which effectively abolishes the ability of RANTES to recruit cells in vivo. These crystallographic structures provide a description of the molecular interaction of a chemokine with glycosaminoglycans. PMID- 15530373 TI - Keeping up with bioterrorism fears. PMID- 15530375 TI - Reeve's stem-cell legacy. PMID- 15530376 TI - Steven Pinker. PMID- 15530377 TI - Dolphin cognition. PMID- 15530379 TI - The elusive flippases. PMID- 15530380 TI - Human cilia proteome contains homolog of zebrafish polycystic kidney disease gene qilin. PMID- 15530381 TI - DNA replication: stalling a fork for imprinting and switching. AB - Mating-type switching in fission yeast has long been known to be directed by a DNA 'imprint'. This imprint has now been firmly characterized as a protected site specific and strand-specific nick. New work also links the widely conserved Swi1 Swi3 complex to the protection of stalled replication forks in general. PMID- 15530382 TI - Olfaction: attracting both sperm and the nose. AB - Odorant receptor genes are expressed not only in the nose but also in testes, where they have been hypothesized to play a role in sperm chemotaxis. New data demonstrate that human odorant receptor hOR 17-4 may play similar roles in both tissues, lending support to the idea that chemical attraction is important for reproduction. PMID- 15530383 TI - Kinetochores: if you build it, they will come. AB - Successful mitosis depends critically on the segregation of chromosomes by kinetochore microtubules. A recent paper describes a conserved protein network from Caenorhabditis elegans that is composed of three classes of molecules, each of which contributes uniquely to the building of the kinetochore-microtubule attachment site. PMID- 15530384 TI - Proteolysis: Adaptor, adaptor, catch me a catch. AB - The ClpXP protease of bacteria can degrade a wide variety of proteins while maintaining remarkable substrate selectivity. New work in Escherichia coli implicates adaptor proteins in enhancing substrate selectivity and regulating the flow of substrates to cellular proteases. PMID- 15530385 TI - Language: Life without numbers. AB - If your language did not have words for numbers, would you be able to think about numeric quantities? An Amazonian culture where number words are limited to one, two and many has provided new insights to the interaction between thought and language. PMID- 15530386 TI - Plant signaling: notes from the underground. AB - New studies combining genetic and grafting approaches in Arabidopsis provide evidence that a carotenoid derivative is a novel plant signaling molecule through which roots can influence shoot branching and leaf development. PMID- 15530387 TI - Actin motility: staying on track takes a little more effort. AB - The pigment cells of amphibians and fish have provided excellent models for understanding how actin and microtubule motors move organelles along the complex trailway of filaments in the cytoplasm. Recent work provides more evidence that these motors are engaged in a continuous tug-of-war. PMID- 15530388 TI - Cell signalling: IP3 receptors channel calcium into cell death. AB - There is substantial evidence that Ca2+ fluxes occur during most forms of apoptosis, and that inhibiting such fluxes protects cells from death. IP3 receptors--ligand-gated channels that release Ca2+ from intracellular stores--are emerging as key sites for regulation by pro- and anti-apoptotic factors. PMID- 15530389 TI - MEC-2 is recruited to the putative mechanosensory complex in C. elegans touch receptor neurons through its stomatin-like domain. AB - BACKGROUND: The response to gentle body touch in C. elegans requires a degenerin channel complex containing four proteins (MEC-2, MEC-4, MEC-6, and MEC-10). The central portion of the integral membrane protein MEC-2 contains a stomatin-like region that is highly conserved from bacteria to mammals. The molecular function of this domain in MEC-2, however, is unknown. RESULTS: Here, we show that MEC-2 colocalizes with the degenerin MEC-4 in regular puncta along touch receptor neuron processes. This punctate localization requires the other channel complex proteins. The stomatin-like region of MEC-2 interacts with the intracellular cytoplasmic portion of MEC-4. Missense mutations in this region that destroy the interaction also disrupt the punctate localization and degenerin-regulating function of MEC-2. Missense mutations outside this region apparently have no effect on the punctate localization but significantly reduce the regulatory effect of MEC-2 on the MEC-4 degenerin channel. A second stomatin-like protein, UNC-24, colocalizes with MEC-2 in vivo and coimmunoprecipitates with MEC-2 and MEC-4 in Xenopus oocytes; unc-24 enhances the touch insensitivity of temperature sensitive alleles of mec-4 and mec-6. CONCLUSION: Two stomatin homologs, MEC-2 and UNC-24, interact with the MEC-4 degenerin through their stomatin-like regions, which act as protein binding domains. At least in the case of MEC-2, this binding allows its nonstomatin domains to regulate channel activity. Stomatin-like regions in other proteins may serve a similar protein binding function. PMID- 15530390 TI - VPEgamma exhibits a caspase-like activity that contributes to defense against pathogens. AB - BACKGROUND: Caspases are a family of aspartate-specific cysteine proteases that play an essential role in initiating and executing programmed cell death (PCD) in metazoans. Caspase-like activities have been shown to be required for the initiation of PCD in plants, but the genes encoding those activities have not been identified. VPEgamma, a cysteine protease, is induced during senescence, a form of PCD in plants, and is localized in precursor protease vesicles and vacuoles, compartments associated with PCD processes in plants. RESULTS: We show that VPEgamma binds in vivo to a general caspase inhibitor and to caspase-1 specific inhibitors, which block the activity of VPEgamma. A cysteine protease inhibitor, cystatin, accumulates to 20-fold higher levels in vpegamma mutants. Homologs of cystatin are known to suppress hypersensitive cell death in plant and animal systems. We also report that infection with an avirulent strain of Pseudomonas syringae results in an increase of caspase-1 activity, and this increase is partially suppressed in vpegamma mutants. Plants overexpressing VPEgamma exhibit a greater amount of ion leakage during infection with P. syringae, suggesting that VPEgamma may regulate cell death progression during plant-pathogen interaction. VPEgamma expression is induced after infection with P. syringae, Botrytis cinerea, and turnip mosaic virus, and knockout of VPEgamma results in increased susceptibility to these pathogens. CONCLUSIONS: We conclude that VPEgamma is a caspase-like enzyme that has been recruited in plants to regulate vacuole-mediated cell dismantling during cell death, a process that has significant influence in the outcome of a diverse set of plant-pathogen interactions. PMID- 15530391 TI - Increased neuronal excitability, synaptic plasticity, and learning in aged Kvbeta1.1 knockout mice. AB - BACKGROUND: Advancing age is typically accompanied by deficits in learning and memory. These deficits occur independently of overt pathology and are often considered to be a part of "normal aging." At the neuronal level, normal aging is known to be associated with numerous cellular and molecular changes, which include a pronounced decrease in neuronal excitability and an altered induction in the threshold for synaptic plasticity. Because both of these mechanisms (neuronal excitability and synaptic plasticity) have been implicated as putative cellular substrates for learning and memory, it is reasonable to propose that age related changes in these mechanisms may contribute to the general cognitive decline that occurs during aging. RESULTS: To further investigate the relationship between aging, learning and memory, neuronal excitability, and synaptic plasticity, we have carried out experiments with aged mice that lack the auxiliary potassium channel subunit Kvbeta1.1. In aged mice, the deletion of the auxiliary potassium channel subunit Kvbeta1.1 resulted in increased neuronal excitability, as measured by a decrease in the post-burst afterhyperpolarization. In addition, long-term potentiation (LTP) was more readily induced in aged Kvbeta1.1 knockout mice. Finally, the aged Kvbeta1.1 mutants outperformed age matched controls in the hidden-platform version of the Morris water maze. Interestingly, the enhancements in excitability and learning were both sensitive to genetic background: The enhanced learning was only observed in a genetic background in which the mutants exhibited increased neuronal excitability. CONCLUSIONS: Neuronal excitability is an important determinant of both synaptic plasticity and learning in aged subjects. PMID- 15530392 TI - Visualizing long-range movement of the morphogen Xnr2 in the Xenopus embryo. AB - One way in which cells acquire positional information during embryonic development is by measuring the local concentration of a signaling factor, or morphogen, that is secreted by an organizing center . The ways in which morphogen gradients are established, particularly in vertebrates, remain obscure, although various suggestions have been made for the mechanisms by which signaling molecules traverse fields of cells. These include simple diffusion, "cytonemes", filopodia, "argosomes", and "transcytosis". In this study, we use a functional EGFP-tagged ligand to visualize long-range signaling in the Xenopus embryo in real time. Our results show that the TGF-beta family member Xnr2 is secreted efficiently from embryonic cells, and a new method of tissue recombination allows us to investigate the way in which the morphogen traverses multiple cell diameters. This reveals that Xnr2 exerts long-range effects by diffusing rapidly through the extracellular milieu of nonexpressing cells. No evidence has been obtained for long-range signaling through cytonemes, filopodia, argosomes, or transcytosis. In demonstrating that long-range signaling in the early Xenopus embryo occurs by diffusion rather than by these alternative routes, our results suggest that different morphogens in different developmental contexts use different means of transport. PMID- 15530393 TI - A programmed strand-specific and modified nick in S. pombe constitutes a novel type of chromosomal imprint. AB - The sexual locus mat1, in the fission yeast Schizosaccharomyces pombe, efficiently switches between the two mating types, P and M, by a process similar to gene conversion, using the silent mat2-P and mat3-M loci, respectively, as donors of the P and M genetic information . It has been proposed that an asymmetrically inherited, site- and strand-specific imprint at mat1 initiates the mating-type switching process . The molecular nature of the imprint is controversial; it was initially described as a double-strand break and then as a single-strand lesion or a strand-specific, alkali-labile modification . Here, we use E. coli DNA ligase in vitro to demonstrate that the imprint is a nick with no resection of nucleotides. By using ligation-mediated PCR, we show that the nick contains 3'OH and 5'OH unphosphorylated termini resistant to RNase treatments. This nonmutational mark on one of the DNA strands provides the first example of a novel type of imprint. PMID- 15530394 TI - Identification of bacterial muramyl dipeptide as activator of the NALP3/cryopyrin inflammasome. AB - Activation of caspase-1 and subsequent processing and secretion of the pro inflammatory cytokine IL-1beta is triggered upon assembly of the inflammasome complex. It is generally believed that bacterial lipopolysaccharides (LPS) are activators of the inflammasome through stimulation of Toll-like receptor 4 (TLR4). Like TLRs, NALP3/Cryopyrin, which is a key component of the inflammasome, contains Leucine-Rich-Repeats (LRRs). LRRs are frequently used to sense bacterial components, thus raising the possibility that bacteria directly activate the inflammasome. Here, we show that bacterial peptidoglycans (PGN), but surprisingly not LPS, induce NALP3-mediated activation of caspase-1 and maturation of proIL 1beta. Activation is independent of TLRs because the PGN degradation product muramyl dipeptide (MDP), which is not sensed by TLRs, is the minimal-activating structure. Macrophages from a patient with Muckle-Wells syndrome, an autoinflammatory disease associated with mutations in the NALP3/Cryopyrin gene, show increased IL-1beta secretion in the presence of MDP. The activation of the NALP3-inflammasome by MDP may be the basis of the potent adjuvant activity of MDP. PMID- 15530395 TI - The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity. AB - The ABC model of flower organ identity is widely recognized as providing a framework for understanding the specification of flower organs in diverse plant species. Recent studies in Arabidopsis thaliana have shown that three closely related MADS-box genes, SEPALLATA1 (SEP1), SEP2 and SEP3, are required to specify petals, stamens, and carpels because these organs are converted into sepals in sep1 sep2 sep3 triple mutants. Additional studies indicate that the SEP proteins form multimeric complexes with the products of the B and C organ identity genes. Here, we characterize the SEP4 gene, which shares extensive sequence similarity to and an overlapping expression pattern with the other SEP genes. Although sep4 single mutants display a phenotype similar to that of wild-type plants, we find that floral organs are converted into leaf-like organs in sep1 sep2 sep3 sep4 quadruple mutants, indicating the involvement of all four SEP genes in the development of sepals. We also find that SEP4 contributes to the development of petals, stamens, and carpels in addition to sepals and that it plays an important role in meristem identity. These and other data demonstrate that the SEP genes play central roles in flower meristem identity and organ identity. PMID- 15530396 TI - Eg5 causes elongation of meiotic spindles when flux-associated microtubule depolymerization is blocked. AB - In higher eukaryotes, microtubules (MT) in both halves of the mitotic spindle translocate continuously away from the midzone in a phenomenon called poleward microtubule flux. Because the spindle maintains constant length and microtubule density, this microtubule translocation must somehow be coupled to net MT depolymerization at spindle poles. The molecular mechanisms underlying both flux associated translocation and flux-associated depolymerization are not well understood, but it can be predicted that blocking pole-based destabilization will increase spindle length, an idea that has not been tested in meiotic spindles. Here, we show that simultaneous addition of two pole-disrupting reagents p50/dynamitin and a truncated version of Xklp2 results in continuous spindle elongation in Xenopus egg extracts, and we quantitatively correlate this elongation rate with the poleward translocation of stabilized microtubules. We further use this system to demonstrate that this poleward translocation requires the activity of the kinesin-related protein Eg5. These results suggest that Eg5 is responsible for flux-associated MT translocation and that dynein and Xklp2 regulate flux-associated microtubule depolymerization at spindle poles. PMID- 15530397 TI - Migratory birds use head scans to detect the direction of the earth's magnetic field. AB - Night-migratory songbirds are known to use a magnetic compass , but how do they detect the reference direction provided by the geomagnetic field, and where is the sensory organ located? The most prominent characteristic of geomagnetic sensory input, whether based on visual patterns or magnetite-mediated forces , is the predicted symmetry around the north-south or east-west magnetic axis. Here, we show that caged migratory garden warblers perform head-scanning behavior well suited to detect this magnetic symmetry plane. In the natural geomagnetic field, birds move toward their migratory direction after head scanning. In a zero magnetic field , where no symmetry plane exists, the birds almost triple their head-scanning frequency, and the movement direction after a head scan becomes random. Thus, the magnetic sensory organ is located in the bird's head, and head scans are used to locate the reference direction provided by the geomagnetic field. PMID- 15530398 TI - Inscuteable mRNA localization is dynein-dependent and regulates apicobasal polarity and spindle length in Drosophila neuroblasts. AB - Drosophila neuroblasts undergo asymmetric divisions along the apicobasal axis to produce two daughter cells of unequal size and different developmental fate. Inscuteable (Insc) protein functions as part of an apically localized complex to coordinate orientation of the mitotic spindle and basal sorting of cell fate determinants. insc mRNA transcripts also localize apically in neuroblasts, yet the mechanism underpinning this process and its developmental significance are unknown. Here, we show that the Egalitarian (Egl)/Bicaudal-D (BicD)/dynein mRNA transport machinery mediates apical localization of insc mRNA transcripts in neuroblasts, and we provide evidence that insc localization is required for efficient apical targeting of Insc protein. egl and BicD mutant neuroblasts display defects in apicobasal polarity, which is consistent with apical Insc activity being reduced. Also, we observe shortened mitotic spindles at metaphase in egl, BicD, and insc mutant neuroblasts and demonstrate a previously unknown, dose-dependent requirement for Insc in augmenting metaphase spindle length. We conclude that localization of insc mRNA transcripts in neuroblasts confers maximal levels of apical Insc activity, which is required for accurate control of metaphase spindle length, division orientation, and asymmetric cell division. PMID- 15530399 TI - The Drosophila microtubule-associated protein mini spindles is required for cytoplasmic microtubules in oogenesis. AB - The XMAP215/TOG family of proteins is a closely related set of MAPs (microtubule associated proteins) found in animals, yeast, and plants . In yeast and animal cells, the XMAP215/TOG proteins are required for both mitosis and meiosis. Although effects of XMAP215/TOG proteins on cytoplasmic microtubules have not previously been shown in animal cells, in plants the Arabidopsis family member MOR1 is required for the organization of cortical microtubule arrays . The Drosophila family member, encoded by the mini spindles (msps) gene, is maternally expressed and loaded into the egg, where it is an essential component of meiotic and mitotic spindles . Here we show that msps is also required during oogenesis for the structure and function of cytoplasmic microtubules. Localization of bicoid (bcd) mRNA in the oocyte is a microtubule-mediated event . We show that bcd RNA localization is defective in msps mutants. We also identify defects in cytoplasmic microtubules in both the germ and follicle cells of mutant ovaries and determine the expression pattern of msps mRNA and protein in developing egg chambers. Our findings reveal a new role for msps in cell patterning and raise the possibility that other family members may perform similar functions. PMID- 15530400 TI - Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase. AB - With a single microtubule attachment, budding-yeast kinetochores provide an excellent system for understanding the coordinated linkage to dynamic microtubule plus ends for chromosome oscillation and positioning. Fluorescent tagging of kinetochore proteins indicates that, on average, all centromeres are clustered, distinctly separated from their sisters, and positioned equidistant from their respective spindle poles during metaphase. However, individual fluorescent chromosome markers near the centromere transiently reassociate with their sisters and oscillate from one spindle half to the other. To reconcile the apparent disparity between the average centromere position and individual centromere proximal markers, we utilized fluorescence recovery after photobleaching to measure stability of the histone-H3 variant Cse4p/CENP-A. Newly synthesized Cse4p replaces old protein during DNA replication. Once assembled, Cse4-GFP is a physically stable component of centromeres during mitosis. This allowed us to follow centromere dynamics within each spindle half. Kinetochores remain stably attached to dynamic microtubules and exhibit a low incidence of switching orientation or position between the spindle halves. Switching of sister chromatid attachment may be contemporaneous with Cse4p exchange and early kinetochore assembly during S phase; this would promote mixing of chromosome attachment to each spindle pole. Once biorientation is attained, centromeres rarely make excursions beyond their proximal half spindle. PMID- 15530402 TI - Correlations between genomic GC levels and optimal growth temperatures are not 'robust'. AB - Musto et al. [FEBS Lett. 573 (2004) 73] studied the correlations between GC levels and optimal growth temperatures in 20 prokaryotic families. They reported that positive correlations are generally observed, and many of these are significant. Here, we have shown that these correlations are not "robust," i.e., correlation coefficients and/or significance of correlations can be considerably influenced by exclusion of very few (even as small as one) species from each dataset. The sensitivity of correlations is assumed as a result of high levels of bias in the family datasets. We concluded that solely based on these data, one cannot establish that GC contents of prokaryotic genomes increase as a result of growth temperature increments. PMID- 15530401 TI - Proteolysis contributes to the exclusive centromere localization of the yeast Cse4/CENP-A histone H3 variant. AB - Kinetochores are the specialized protein structures that form on centromeric DNA and direct chromosome segregation. It is critical that all chromosomes assemble a single kinetochore every cell cycle. One hallmark of all eukaryotic kinetochores is CENP-A, an essential centromeric histone H3 (CenH3) variant. Overexpression of CENP-A causes mislocalization to euchromatin, which could lead to deleterious consequences because CENP-A overexpression is associated with colorectal cancer . Although CENP-A protein levels are important for genomic stability, little is known about the mechanisms of CenH3 regulation. Here, we show that the levels of the budding yeast CenH3, Cse4, are regulated by ubiquitin-proteasome-mediated proteolysis. Because mutation of all Cse4 lysine residues did not completely stabilize the protein, we isolated a dominant lethal mutant, CSE4-351, that was stable. The Cse4-351 protein localized to euchromatin, suggesting that proteolysis prevents CenH3 euchromatic localization. When wild-type Cse4 was fused to a degron signal, the soluble Cse4 protein was rapidly degraded, but the centromere bound Cse4 was stable, indicating that centromere localization protects Cse4 from degradation. Taken together, these data identify proteolysis as one mechanism that contributes to the restricted centromere localization of the yeast CenH3. PMID- 15530403 TI - Abeta N-terminal-end specific antibody reduced beta-amyloid in Alzheimer-model mice. AB - Alzheimer's disease (AD) is a neurodegenerative disease with memory dysfunction that is causing serious medical problems in modern society. For the fundamental treatment of AD, an amyloid beta protein (Abeta) vaccine is considered to be the most potent candidate. To cure AD, we developed Abeta N-terminal-end specific monoclonal antibody named 82E1, which does not cross-react with full-length Abeta precursor. Passive intraperitoneal administration of 82E1 markedly reduced total plaque area (Abeta burden) in the Tg2576 mouse brains. This was confirmed by the ELISA measurement of insoluble Abeta in the brain homogenates. The density of diffuse plaques, which increases in the late stage, was markedly reduced by the administration of 82E1, suggesting that the reduction of the Abeta burden was due to the prevention of newly developed diffuse plaques. Above results provide an insight into the further therapeutic intervention in AD with few adverse effects. PMID- 15530404 TI - Copper-mediated homo-dimerisation for the HAH1 metallochaperone. AB - The HAH1 metallochaperone is a key protein implicated in copper homeostasis in human cells. Using as solid-phase based assay completed with Biacore studies, we provided evidence that HAH1 forms homo-dimers in the presence of copper. Biacore analysis allowed us to determine the kinetic parameters of this interaction, characterised by an apparent affinity constant of 6muM. Moreover, we demonstrated that copper-loaded HAH1 interacts independently with each of the six individual metal-binding domains of the copper-translocating Menkes ATPase. Finally, the homo-dimerisation of the metallochaperone was confirmed in living cells by using fluorescence resonance energy transfer. Results have been discussed in the context of intracellular copper control. PMID- 15530405 TI - Apelin is a novel angiogenic factor in retinal endothelial cells. AB - There has been much focus recently on the possible functions of apelin, an endogenous ligand for the orphan G-protein-coupled receptor APJ, in cardiovascular and central nervous systems. We report a new function of apelin as a novel angiogenic factor in retinal endothelial cells. The retinal endothelial cell line RF/6A highly expressed both apelin and APJ transcripts, while human umbilical venous endothelial cells (HUVECs) only expressed apelin mRNA. In accordance with these observations, apelin at concentrations of 1 pM-1 microM significantly enhanced migration, proliferation, and capillary-like tube formation of RF/6A cells, but not those of HUVECs, whereas VEGF stimulates those parameters of both cell types. In vivo Matrigel plug assay for angiogenesis, the inclusion of 1 nM apelin in the Matrigel resulted in clear capillary-like formations with an increase of hemoglobin content in the plug. This is the first report showing that apelin is an angiogenic factor in retinal endothelial cells. PMID- 15530406 TI - Detection of oligomerisation and substrate recognition sites of small heat shock proteins by peptide arrays. AB - Small heat shock proteins (sHsps) form large oligomers that are characterised by their dynamic behaviour, e.g., complex disassembly/reassembly and extensive subunit exchange. These processes are interrelated with sHsp/substrate interaction. sHsps bind a broad spectrum of unrelated substrate proteins under denaturing conditions. Detailed knowledge about the binding process and regions critical for sHsp/substrate interaction is missing. In this study, we screened cellulose-bound peptide spot libraries derived from a bacterial sHsp and the model-substrate citrate synthase to detect oligomerisation and substrate interaction sites, respectively. In line with previous results, it was demonstrated that multiple contacts involving the N- and C-terminal extensions and the central alpha-crystallin domain are required for oligomerisation. Incubation of the citrate synthase membrane with sHsps revealed a putative substrate interaction site. A soluble peptide with the sequence RTKYWELIYEDCMDL (CS(191-205)) corresponding to that site inhibited chaperone activity of sHsps, presumably by blocking their substrate-binding sites. PMID- 15530407 TI - Regulation of factors controlling angiogenesis in liver development: a role for PEDF in the formation and maintenance of normal vasculature. AB - PEDF and VEGF are important inhibitors and promoters of angiogenesis, and the ratio between the two is an important indicator in many neovascular diseases. In mouse liver PEDF and VEGF(165) were co-expressed at very early stages of liver development and their expression increased as liver embryogenesis progressed, suggesting that PEDF and VEGF are both crucial to vasculogenesis as well. VEGF(189) only appears at the P0 stage in liver organogenesis and is maintained at high levels thereafter. PEDF and the two VEGF isoforms are synthesized by fresh and cultured hepatocytes. Expression of VEGF(121) and overexpression of VEGF(165) were only seen in HepG2, a well-characterized hepatocellular carcinoma line. The results suggest that hepatic vascular architecture is under the control of both PEDF and VEGF, and that VEGF(165) and VEGF(189) have distinct functions in normal vascular development of the liver. The VEGF isoforms 121 and 189 may be key regulators of increased vascularity and progression of hepatocellular carcinoma, one of the most common malignant tumors, and may be of prognostic significance for this tumor. PMID- 15530408 TI - Homology modeling and S(N)2 displacement reaction of fluoroacetate dehalogenase from Burkholderia sp. FA1. AB - Fluoroacetate dehalogenase (EC 3.8.1.3) catalyzes the dehalogenation of fluoroacetate and other haloacetates. In order to investigate the relation between the structure and the function, and understand the reaction mechanism of the enzyme, a 3D model of fluoroacetate dehalogenase FAc-DEX FA1 was built by homology-based modeling. The 3D model was optimized by unconstrained molecular dynamics simulation. Furthermore, the optimized 3D model was assessed by comparison of specific properties with two known protein structures. From the final 3D model, we find that the main residues involved in the active site in FAc DEX FA1 were Phe34, Trp148, Tyr147, Tyr212, Asp104, and His271; especially Asp104 was the key nucleophilic residue in substrate binding. A reaction model including Asp104 and the substrate fluoroacetate was then constructed and used to characterize explicit enzymatic reactions. In order to further illustrate catalytic properties, the equilibrium geometries, energies, and frequencies of stationary points (reactants, products, and transition states) of the reaction model were calculated at the B3LYP/6-31G level of theory in both gas phase and solution. The results showed that the reaction in gas was dynamically more favorable than in solution. PMID- 15530409 TI - Creation and characteristics of unnatural CysHis(3)-type zinc finger protein. AB - To investigate the properties of unnatural zinc finger peptides with CysHis(3) type ligand combinations, the HCHH- and CHHH-type zinc finger proteins (zf(HCHH) and zf(CHHH), respectively) were created by mutating Cys to His in the Cys(2)His(2)-type zinc finger of the transcription factor Sp1 (zf(CCHH)). The CD measurements clearly show that the single-finger CysHis(3)-type zinc finger peptides (zf(HCHH)f2 and zf(CHHH)f2) are folded by complexation with Zn(II). From the gel mobility shift assays, the CysHis(3)-type zinc finger proteins (zf(HCHH) and zf(CHHH)) evidently bind to the GC-box DNA, though the DNA binding affinity is lower than that of the wild CCHH-type zinc finger protein. Furthermore, the zf(HCHH)f2 and zf(CHHH)f2 peptides catalyze the hydrolysis of the 4-nitrophenyl acetate in contrast to the catalytically inactive zf(CCHH) peptide. This is the first study of the CysHis(3)-type zinc finger proteins and also provides useful information for redesigning artificial metalloproteins. PMID- 15530410 TI - Aggresome formation and pharmacogenetics: sulfotransferase 1A3 as a model system. AB - A common cause for pharmacogenetic alteration in drug response is genetic variation in encoded amino acid sequence. We have used the catecholamine and drug metabolizing enzyme sulfotransferase (SULT)1A3 to create an artificial model system to study mechanisms-especially possible aggresome formation-by which genetic alteration in amino acid sequence might influence function. Specifically, we created a double variant SULT1A3 allozyme that included the naturally occurring Asn234 polymorphism plus an additional Trp172Arg mutation. Analysis of the SULT1A3 X-ray crystal structure had indicated that the Trp172Arg mutation might destabilize the protein's structure. Expression of SULT1A3 Arg172,Asn234 in COS-1 cells resulted in undetectable enzyme activity and a virtual lack of enzyme protein. Rabbit reticulocyte lysate degradation studies showed that the double variant allozyme was degraded much more rapidly than was wild type SULT1A3 by a ubiquitin-proteasome-dependent process. In addition, after expression in COS-1 cells, the double variant allozyme localized to aggresomes, a process not previously described or studied in pharmacogenetics. Therefore, the alteration of only one or two amino acids can lead to decreased levels of protein as a result of both aggresome formation and accelerated degradation. The possible role of aggresome formation in pharmacogenetics should be evaluated in naturally occurring systems with inherited alteration in encoded amino acid sequence. PMID- 15530411 TI - Oxidation of low density lipoproteins by myeloperoxidase at the surface of endothelial cells: an additional mechanism to subendothelium oxidation. AB - The present paradigm of atherogenesis proposes that low density lipoproteins (LDL) are trapped in subendothelial space of the vascular wall where they are oxidized. Myeloperoxidase (MPO) plays a key role in oxidative damage. We propose that LDL oxidation by myeloperoxidase (Mox-LDL) could occur at the surface of the endothelial cells and not restricted to the subendothelial space. The triad constituted by endothelial cells, circulating LDL and MPO in close interaction, constitutes a synergic mechanism for the genesis of Mox-LDL. PMID- 15530412 TI - The Xenopus laevis morphogenetic factor, tumorhead, causes defects in polarized growth and cytokinesis in the fission yeast, Schizosaccharomyces pombe. AB - Tumorhead (TH) is a maternally expressed gene product that regulates neural tube morphogenesis in the amphibian, Xenopus laevis. Here we describe the effects of TH expression in the rod-shaped fission yeast, Schizosaccharomyces pombe. Expression of TH in S. pombe resulted in severe morphological defects, including ovoid, bottle-shaped, and enlarged cells. Multi-septated cells were also observed in TH expressing cultures, indicating that TH is inhibitory to a process required for the completion of cytokinesis. TH expression caused significant actin and microtubule cytoskeletal defects, including depolarization of the cortical F actin cytoskeleton and increased microtubule formation. Immunostaining experiments showed that TH is localized to the cell cortex, cell tips, and septum in S. pombe cells. Localization of TH to the cell cortex was dependent on the S. pombe PAK homolog, Shk1. Moreover, TH expression was inhibitory to the growth of a mutant defective in Shk1 function, suggesting that TH may interact with a component(s) of a PAK-mediated morphogenetic regulatory pathway in S. pombe. Taken together, our findings demonstrate that S. pombe may be a useful model organism for identifying potential TH interacting factors. PMID- 15530413 TI - Inactivated SARS-CoV vaccine elicits high titers of spike protein-specific antibodies that block receptor binding and virus entry. AB - The only severe acute respiratory syndrome (SARS) vaccine currently being tested in clinical trial consists of inactivated severe acute respiratory syndrome associate coronavirus (SARS-CoV). However, limited information is available about host immune responses induced by the inactivated SARS vaccine. In this study, we demonstrated that SARS-CoV inactivated by beta-propiolactone elicited high titers of antibodies in the immunized mice and rabbits that recognize the spike (S) protein, especially the receptor-binding domain (RBD) in the S1 region. The antisera from the immunized animals efficiently bound to the RBD and blocked binding of RBD to angiotensin-converting enzyme 2, the functional receptor on the susceptible cells for SARS-CoV. With a sensitive and quantitative single-cycle infection assay using pseudovirus bearing the SARS-CoV S protein, we demonstrated that mouse and rabbit antisera significantly inhibited S protein-mediated virus entry with mean 50% inhibitory titers of 1:7393 and 1:2060, respectively. These data suggest that the RBD of S protein is a major neutralization determinant in the inactivated SARS vaccine which can induce potent neutralizing antibodies to block SARS-CoV entry. However, caution should be taken in using the inactivated SARS-CoV as a vaccine since it may also cause harmful immune and/or inflammatory responses. PMID- 15530414 TI - Cartilage-derived morphogenetic protein-1 promotes the differentiation of mesenchymal stem cells into chondrocytes. AB - Mesenchymal stem cells (MSCs) are able to differentiate into many types of cells including chondrocytes. Transforming growth factor beta1 (TGF-beta1) is very important in the regulation of chondrogenesis. Since cartilage-derived morphogenetic protein-1 (CDMP-1) belongs to the TGF-beta superfamily, we tested whether CDMP-1 plays any role in the regulation of the differentiation of MSCs into chondrocytes using a high density pellet culture system. Based on the histological staining of glycosaminoglycan using toluidine blue dye-binding method we found that CDMP-1 could initiate chondrogenic differentiation of MSCs as did TGF-beta1. However, CDMP-1 was less stimulatory than TGF-beta1. The combination of CDMP-1 and TGF-beta1 synergically induced chondrogenesis of MSCs. This synergic chondrogenic effect of CDMP-1 together with TGF-beta1 was further confirmed by quantification of GAG using dimethylmethylene blue dye-binding assay and immunohistochemical analysis of the expression of cartilage-specific protein collagen II. This study may provide an improved induction approach using MSCs for repairing damaged cartilage. PMID- 15530415 TI - COX-2-selective inhibitor, etodolac, suppresses choroidal neovascularization in a mice model. AB - Cyclooxygenases (COXs) are involved in choroidal neovascularization (CNV). However, the relative contribution of COX-1 and -2 to CNV has not been determined. In this study, the expression of COX-2 was investigated in CNVs in a murine laser-induced model. Subsequently, we found that experimental CNV expressed COX-2, most remarkably around the highly vascularized lesions. To examine the effect of COX-2 inhibition on CNV, etodolac, a non-steroidal anti inflammatory drug with a high COX-2 selectivity, was tested on murine CNV model. The results demonstrated that the intensity of fluorescein leakage from the photocoagulated lesions decreased significantly compared to the control eyes following etodolac administration. The area of CNV lesions, as examined using histological sections and choroidal flatmounts at day 7, demonstrated that the average size of the CNV lesions was significantly reduced in the etodolac-treated eyes compared to the control eyes. Together, our results demonstrated that selective COX-2 inhibition suppresses CNV. PMID- 15530416 TI - Three-base periodicity patterns and self-similarity in whole bacterial chromosomes. AB - It has been reported that in a collection of mRNAs the triplets GhN or RNY had a higher propensity to be separated by either three/six/nine, etc., bases than by two/four/five, etc., bases. This has been called three-base periodicity (TBP). In this work the frequency distribution of distances (FDDs) for all triplets in the Borrelia burgdorferi chromosome and selected triplets in other model sequences were determined. The FDDs produced oscillatory decaying patterns with TBP for most triplets and not only for those encompassed by the above formulas. Furthermore, we also found TBP for di- and mononucleotides. However, TBP was not observed for intergenic regions, sequences with a low content of coding regions or when the coding potential of sequences was disrupted by base shuffling. Excluding closely related species the FDDs between bacterial genomes were different and appeared characteristic of the analyzed genome. FDDs also showed self-similarity, since 1Mb sequences rendered FDDs that were very similar to those for the entire sequence. PMID- 15530417 TI - Interactions within the ubiquitin pathway of Caenorhabditis elegans. AB - The ubiquitin system is a well-conserved and pervasive process for post-synthetic modification of proteins. Three key components of the pathway are required for ubiquitination to occur: the E1 ubiquitin activating enzyme, the E2 ubiquitin conjugating enzyme, and the E3 ubiquitin ligase. There are several different E2 ubiquitin conjugating enzymes and an even greater number of E3 ubiquitin ligases. Interactions between these two groups are critical for substrate ubiquitination. This study reports a two-hybrid analysis of interactions within the ubiquitin system of Caenorhabditis elegans. Forty-three RING finger proteins (presumed E3 ubiquitin ligases) and 14 predicted E2 ubiquitin conjugating enzymes were included in the screen. A total of 31 E2-E3 interactions were uncovered. In addition, the UBC-13 conjugating enzyme was observed to interact with two different E2s, UEV-1 and UBC-1. The interaction of UBC-1 and UBC-13 was confirmed with in vitro ubiquitination reactions. Using NHL-1 as the E3 in the assays, ubiquitination was observed when both UBC-1 and UBC-13 were present but not with either alone. These data imply that some E2s require dimerization in order to function. PMID- 15530418 TI - Molecular modeling and small angle X-ray scattering studies of Hoplosternum littorale cathodic haemoglobin. AB - Considerable interest is currently focused on fish haemoglobins in order to identify the structural basis for their diversity of functional behavior. Hoplosternum littorale is a catfish that presents bimodal gill (water)/gut (air) breathing, which allows this species to survive in waters with low oxygen content. The hemolysate of this fish showed the presence of two main haemoglobins, cathodic and anodic. This work describes structural features analyzed here by integration of molecular modeling with small angle X-ray scattering. Here is described a molecular model for the cathodic haemoglobin in the unliganded and liganded states. The models were determined by molecular modeling based on the high-resolution crystal structure of fish haemoglobins. The structural models for both forms of H. littorale haemoglobin were compared to human haemoglobin. PMID- 15530419 TI - Selective inhibition of enterovirus 71 replication by short hairpin RNAs. AB - RNA interference (RNAi) is a sequence-specific, post-transcriptional process of mRNA degradation induced by small interfering RNA molecules. In this report, RNAi strategy was exploited to treat the infection of enterovirus 71 (EV71), considered as one of the most virulent pathogens that can cause severe complications in the family of Picornaviridae. We developed short hairpin RNA (shRNA) expression plasmids that significantly inhibited viral protein expression in a sequence-specific and dose-dependent fashion after transient transfection in cell cultures. Stable expression of shRNAs in cultured cells exhibited marked viral resistance in every step assessed in the viral replication. Using cytotoxicity of shRNA-expressing cells as a surrogate marker, it was shown that replication of EV71 was specifically attenuated by these plasmid-derived shRNAs, while replications of other related enteroviruses examined were not. These proof of-concept studies demonstrated the feasibility of this approach for the therapy of EV71-associated diseases. PMID- 15530420 TI - Augmentation of antigen-specific immune responses using DNA-fusogenic liposome vaccine. AB - In an attempt to enhance the immunological efficacy of genetic immunization, we investigated a new biological means for delivering antigen gene directly to the cytoplasm via membrane fusion. In this context, we investigated fusogenic liposome (FL) encapsulating DNA as a possible genetic immunization vehicle. RT PCR analysis indicated that a FL could introduce and express encapsulating OVA gene efficiently and rapidly in vitro. Consistent with this observation, an in vitro assay showed that FL-mediated antigen-gene delivery can induce potent presentation of antigen via the MHC class I-dependent pathway. Accordingly, immunization with FL containing the OVA-gene induced potent OVA-specific Th1 and Th2 cytokine production. Additionally, OVA-specific CTL responses and antibody production were also observed in systemic compartments including the spleen, upon immunization with the OVA-gene encapsulating FL. These findings suggest that FL is an effective genetic immunization carrier system for the stimulation of antigen-specific immune responses against its encoding antigen. PMID- 15530421 TI - Using GO-PseAA predictor to predict enzyme sub-class. AB - Enzyme function is much less conserved than anticipated, i.e., the requirement for sequence similarity that implies similarity in enzymatic function is much higher than the requirement that implies similarity in protein structure. This is because the function of an enzyme is an extremely complicated problem that may involve very subtle structural details as well as many other physical chemistry factors. Accordingly, if simply based on the sequence similarity approach, it would hardly get a decent success rate in predicting enzyme sub-class even for a dataset consisting of samples with 50% sequence identity. To cope with such a situation, the GO-PseAA predictor was adopted to identify the sub-class for each of the six main enzyme families. It has been observed that, even for the much more stringent datasets in which none of the enzymes has 25% sequence identity to any others, the overall success rates are 73-95%, suggesting that the GO-PseAA predictor can catch the core features of the statistical samples concerned and may become a useful high throughput tool in proteomics and bioinformatics. PMID- 15530422 TI - Identification and expression of two novel cytochrome P450 genes, belonging to CYP4 and a new CYP331 family, in the polychaete Capitella capitata sp.I. AB - The polychaete Capitella capitata sp.I has a high capacity to metabolize polycyclic aromatic hydrocarbons (PAHs) which are among the most hazardous environmental pollutants with significant biological effects. In the present study, two novel cytochrome P450 (CYP) genes were identified in this species. One was named CYP331A1, the first member of a new family of CYP331, and the other CYP4AT1 is the first member of a new subfamily CYP4AT. Both of these genes are constitutively expressed in the worms and detectable by RT-PCR. The expression of CYP331A1 mRNA was observed to be more sensitive to PAH exposure than CYP4AT1, which indicated that CYP331A1 should play a more important role than CYP4AT1 in PAH metabolism in this species. Considering the importance of C. capitata sp.I in taking up PAH and other organic pollutants from contaminated marine sediments with the potential for subsequent food-chain transfer, our results are important for understanding the molecular basis of biotransformation and detoxification in this invertebrate, and also have evolutionary significance for understanding the diversity and history of the CYP superfamily. PMID- 15530423 TI - A novel hIL-6 antagonist peptide from computer-aided design contributes to suppression of apoptosis in M1 cells. AB - Based on the complex crystal structure of human interleukin-6 (hIL-6) and its receptor (hIL-6R), a novel hIL-6 antagonist peptide (named PT) was designed using computer-guided design method. Dealing with molecular docking and molecular dynamics methods, the interaction between PT and hIL-6R was analyzed. The theoretical studies showed that PT possessed very high affinity to hIL-6R and offered a practical means of imposing long-term blockade of hIL-6 activity in vivo. This effect was examined due to growth arrest and apoptosis induced by hIL 6 in myeloblastic cell line M1 cells in a dose-dependent manner. The findings demonstrate that PT could also act as an excellent antagonist candidate for the induction of growth arrest and apoptosis. Furthermore, murine M1 myeloid cell line, which was induced by the physiological inducer hIL-6 to undergo apoptosis and growth arrest, could be used as a subtle model system to test hIL-6 antagonist. PMID- 15530424 TI - A new putative cyclic nucleotide-gated channel gene, cng-3, is critical for thermotolerance in Caenorhabditis elegans. AB - Cyclic nucleotide-gated (CNG) channels encoded by tax-4 and tax-2 genes are required for chemo- and thermo-sensation in Caenorhabditis elegans. Here we report the identification and the characterization of cng-3, a new CNG channel gene, found in C. elegans. CNG-3 contains six putative transmembrane regions and a cyclic nucleotide-binding domain that show high homology with CNG channels of higher animals as well as TAX-4. The expression of cng-3 is detected from early stages in worm development and restricted in five sensory neurons of amphid including AFD neuron. While a cng-3 null mutant displays normal chemotaxis to volatile odorants, the mutant worms exhibit impaired thermal tolerance. These results indicate that CNG-3, a new member of CNG channel subunits, may play a critical role in sensation or response of thermal stress in C. elegans. PMID- 15530425 TI - ACE inhibition lowers angiotensin II-induced chemokine expression by reduction of NF-kappaB activity and AT1 receptor expression. AB - OBJECTIVE: Angiotensin converting enzyme (ACE) inhibitors significantly improve survival in patients with atherosclerosis. Although ACE inhibitors reduce local angiotensin II (AngII) formation, serine proteases form AngII to an enormous amount independently from ACE. Therefore, our study concentrates on the effect of the ACE-inhibitor ramiprilat on chemokine release, AngII receptor (ATR) expression, and NF-kappaB activity in monocytes stimulated with AngII. METHODS AND RESULTS: AngII-induced upregulation of IL-8 and MCP-1 protein and RNA in monocytes was inhibited by the AT1R-blocker losartan, but not by the AT2R-blocker PD 123.319. Ramiprilat dose-dependently suppressed AngII-induced upregulation of IL-8 and MCP-1. The suppressive effect of ramiprilat on AngII-induced chemokine production and release was in part caused by downregulation of NF-kappaB, but more by a selective and highly significant reduced expression of AT1 receptors as shown in monocytes and endothelial cells. CONCLUSION: In our study we demonstrated for the first time that ramiprilat reduced expression of AT1R in monocytes and endothelial cells. In addition, ramiprilat downregulated NF-kappaB activity and thereby reduced the AngII-induced release of IL-8 and MCP-1 in monocytes. This antiinflammatory effect, at least in part, may contribute to the clinical benefit of the ACE inhibitor in the treatment of coronary artery disease. PMID- 15530426 TI - Region 752-761 of STAT3 is critical for SRC-1 recruitment and Ser727 phosphorylation. AB - STAT3 regulates many target genes in response to cytokines and growth factors. To study the mechanisms of STAT3-dependent transcription, we established several cell lines in which HepG2-STAT3-knockdown cells were reconstituted with a variety of STAT3 mutants. Using these cell lines, we found that truncated STAT3(1-750), but not STAT3(1-761), could not recruit SRC-1/NcoA-1 and was not phosphorylated on Ser727. Furthermore, mutation of STAT3 L755 and F757 to alanines caused the loss of STAT3-dependent SRC-1 recruitment, leaving Ser727 phosphorylation intact. Consistent with this, the STAT3-L755A/F757A mutant showed no increase in acetylated histone H3 at Lys14 and a decreased level of RNA polymerase II recruited to the target gene promoter, although p300 recruitment and histone H4 acetylation were intact. This mutant also lost responsiveness to co-expressed SRC 1. Thus, the conserved STAT3 region from 752 to 761, called STAT3 CR2, plays critical roles in STAT3-dependent transcription by recruiting SRC-1 and allowing Ser727 phosphorylation. PMID- 15530427 TI - Gender difference in serum bisphenol A levels may be caused by liver UDP glucuronosyltransferase activity in rats. AB - Gender difference in human bisphenol A (BPA) concentrations was revealed by determining serum BPA. We studied the serum concentrations and the metabolism of BPA in rats by an HPLC system. Rat serum BPA concentrations were significantly higher in males (24.9+/-7.38 ng/ml, P=0.026, n=10) than in females (8.27+/-3.11 ng/ml, n=10), as in humans. The resultant enzyme reaction products of BPA glucuronidation in the rat liver microsomes fraction were analyzed by an HPLC system. The ratio of BPA glucuronidation in the microsome reaction was significantly higher (P=0.015) in female than in male rats. The mRNA expression of UDP-glucuronosyltransferase 2B1 (UGT2B1), an isoform of UGT related to BPA glucuronidation, in the rat liver was analyzed by a real-time quantitative RT PCR. The relative expression level of UGT2B1 mRNA was significantly higher (P<0.001) in female than in male rat livers. The gender difference in serum BPA concentrations may be explained by the difference in clearance based on the UGT activities. PMID- 15530428 TI - Glucagon release is regulated by tyrosine phosphatase and PI3-kinase activity. AB - Since inhibition of protein tyrosine phosphatase (PTPase) activity by peroxovanadate (pV) affects insulin release and phosphorylation of pancreatic islet proteins in the insulin signaling pathway, we studied whether pV also modulates glucagon release. At 3.3mM glucose, pV (0.1-1mM) enhanced glucagon release in a dose-dependent manner in islets of normal Wistar and diabetic GK rats. Arginine-stimulated glucagon responses were higher in GK than in Wistar islets. These responses were inhibited by pV (0.01-0.1mM), also after islet exposure to pertussis toxin (PTX), but were abolished by 1 microM wortmannin. Moreover, in GK but not Wistar islets, wortmannin significantly stimulated basal glucagon secretion (p<0.05) and inhibited arginine-induced glucagon secretion (p<0.001). In In-R1-G9 glucagonoma cells, the inhibitory effect of pV (0.01 mM) on glucagon response to arginine was also observed and paralleled by increased IRS-1 and IRS-2 associated PI3-kinase activity. In conclusion, inhibition of PTPase activity by pV stimulates basal and inhibits arginine-induced glucagon release. The inhibitory effect of 0.01-0.1mM pV seems not to be accounted for by islet peptides acting on PTX sensitive G(i)-proteins. PI3-kinase activity seems to play an important role in pV-induced inhibition of glucagon release. PMID- 15530430 TI - COBRA1 inhibits AP-1 transcriptional activity in transfected cells. AB - Mutations in the breast cancer susceptibility gene (BRCA1) account for a significant proportion of hereditary breast and ovarian cancers. Cofactor of BRCA1 (COBRA1) was isolated as a BRCA1-interacting protein and exhibited a similar chromatin reorganizing activity to that of BRCA1. However, the biological role of COBRA1 remains largely unexplored. Here, we report that ectopic expression of COBRA1 inhibited activator protein 1 (AP-1) transcriptional activity in transfected cells in a dose-dependent manner, whereas reduction of endogenous COBRA1 with a small interfering RNA significantly enhanced AP-1 mediated transcriptional activation. COBRA1 physically interacted with the AP-1 family members, c-Jun and c-Fos, and the middle region of COBRA1 bound to c-Fos. Lack of c-Fos binding site in the COBRA1 completely abolished the COBRA1 inhibition of AP-1 trans-activation. These findings suggest that COBRA1 may directly modulate AP-1 pathway and, therefore, may play important roles in cell proliferation, differentiation, apoptosis, and oncogenesis. PMID- 15530429 TI - Probing biosynthesis of plant polyketides with synthetic N-acetylcysteamine thioesters. AB - Recombinant chalcone synthase (CHS) from Scutellaria baicalensis accepted cinnamoyl diketide-NAC and cinnamoyl-NAC as a substrate, and carried out sequential condensations with malonyl-CoA to produce 2',4',6'-trihydroxychalcone. Steady-state kinetic analysis revealed that the CHS accepted the diketide-NAC with less efficiency, while cinnamoyl-NAC primed the enzyme reaction almost as efficiently as cinnamoyl-CoA. On the other hand, it was for the first time demonstrated that the diketide-NAC was also a substrate for recombinant polyketide reductase (PKR) from Glycyrrhiza echinata, and converted to the corresponding beta-ketohemithioester. Furthermore, by co-action of the CHS and the PKR, the NAC-thioesters were converted to 6'-deoxychalcone in the presence of NADPH and malonyl-CoA. PMID- 15530431 TI - Novel isoforms of the TFIID subunit TAF4 modulate nuclear receptor-mediated transcriptional activity. AB - The transcription factor TFIID consists of TATA-binding protein (TBP) and TBP associated factors (TAFs). TAFs are essential for modulation of transcriptional activity but the regulation of TAFs is complex and many important aspects remain unclear. In this study, we have identified and characterized five novel truncated forms of the TFIID subunit TAF4 (TAF(II)135). Analysis of the mouse gene structure revealed that all truncations were the results of alternative splicing and resulted in the loss of domains or parts of domains implicated in TAF4 functional interactions. Results from transcriptional assays showed that several of the TAF4 isoforms exerted dominant negative effects on TAF4 activity in nuclear receptor-mediated transcriptional activation. In addition, alternative TAF4 isoforms could be detected in specific cell types. Our results indicate an additional level of complexity in TAF4-mediated regulation of transcription and suggest context-specific roles for these new TAF4 isoforms in transcriptional regulation in vivo. PMID- 15530432 TI - Thapsigargin or curcumin does not promote maturation of processing mutants of the ABC transporters, CFTR, and P-glycoprotein. AB - Misprocessed plasma membrane proteins of CFTR and P-glycoprotein (P-gp) are retained in the endoplasmic reticulum (ER) by molecular chaperones. Depletion of the calcium stores in the ER by the SERCA calcium pump inhibitors thapsigargin or curcumin inhibits these interactions and allows the protein to be trafficked to the plasma membrane [Nat. Med. 8 (2002) 485; Science 304 (2004) 600]. We tested this hypothesis by treating various cell lines expressing misprocessed mutants of CFTR or P-gp with thapsigargin or curcumin. Conversion of the immature core glycosylated protein to mature product was detected by immunoblot analysis of whole cell extracts. Mature product was not detected in any of the misprocessed mutants. By contrast, all misprocessed P-gp mutants were rescued by the chemical chaperone/drug substrate cyclosporin A in a dose-dependent manner. These results show that thapsigargin or curcumin is not effective in rescuing misprocessed mutants of P-gp and CFTR. PMID- 15530433 TI - Determination of the transphosphorylation sites of Jak2 kinase. AB - Janus kinases are the key enzymes involved in the initial transmission of signals in response to type I and II cytokines. Activation of the signal begins with the transphosphorylation of Jak kinases. Substrates that give rise to downstream events are recruited to the receptor complex in part by interactions with phosphorylated tyrosines. The identity of many of the phosphotyrosines responsible for recruitment has been elucidated as being receptor-based tyrosines. The ability of Jaks to recruit substrates through their own phosphotyrosines has been demonstrated for tyrosines in the kinase activation loop. Recent studies demonstrate that other tyrosines have implications in regulatory roles of Jak kinase activity. In this study, baculovirus-produced Jak2 was utilized to demonstrate that transphosphorylation of Jak kinases occurs on multiple residues throughout the protein. We demonstrate that among the tyrosines phosphorylated, those in the kinase domain occur as expected, but many other sites are also phosphorylated. The tyrosines conserved in the Jak family are the object of this study, although many of them are phosphorylated, many are not. This result suggests that conservation of tyrosines is perhaps as important in maintaining structure of the Jak family. Additionally, non-Jak family conserved tyrosines are phosphorylated suggesting that the individual Jaks ability to phosphorylated specific tyrosines may influence signals emitting from activated Jaks. PMID- 15530434 TI - Hypertonic induction of aquaporin-1 water channel independent of transcellular osmotic gradient. AB - Aquaporin-1 (AQP1) water channel plays a critical role for water reabsorption in the urinary concentrating mechanism. AQP1 expression in renal cells is upregulated by hypertonicity, but not urea, suggesting the requirement of an osmotic gradient. To investigate whether AQP1 expression is regulated by apical and/or basolateral hypertonicity, murine renal medullary mIMCD-K2 cells grown on permeable support were exposed to hypertonic medium. When the medium on the apical or basolateral membrane side was switched to hypertonic, the transcellular osmotic gradient was dissipated within 8h. Basolateral hypertonicity increased AQP1 expression more than apical hypertonicity. Comparable apical and basolateral hypertonicity without a transcellular hypertonic gradient, however, increased AQP1 expression. Cell surface biotinylation experiments revealed that hypertonicity promoted AQP1 trafficking to both plasma cell membranes. These results indicate that AQP1 expression is predominantly mediated by basolateral hypertonicity but a transcellular osmotic gradient is not necessary for its induction. PMID- 15530435 TI - Elimination of polarity in the carotenoid terminus promotes the exposure of B850 binding sites (Tyr 44, 45) and ANS-mediated energy transfer in LH2 complexes of Rhodobacter sphaeroides. AB - Carotenoids in the peripheral light-harvesting complexes (LH2) of the green mutant (GM309) of Rhodobacter sphaeroides were identified as containing neurosporenes, which lack the polar CH(3)O group, compared to spheroidenes in native-LH2 of R. sphaeroides 601. After LH2 complexes were treated with 1-anilino 8-naphthalene sulfonate (ANS), new energy transfer pathways from ANS or tryptophan to carotenoids were discovered in both native- and GM309-LH2. The carotenoid fluorescence intensity of GM309-LH2 was greater than that of native LH2 when bound with ANS, suggesting that the elimination of polarity in the neurosporene increases the energy transfer from ANS to carotenoid. The fact that two alpha-tyrosines (alpha-Tyr 44, 45, B850-binding sites) in each alpha apoprotein of GM309-LH2 were more easily modified than those of native-LH2 by N acetylimidazole (NAI) indicates that the elimination of polarity in the neurosporene terminus increases the exposure of these sites to solution. PMID- 15530436 TI - Temporal and spatial characteristics of apoptosis in the infarcted rat heart. AB - Following myocardial infarction (MI), tissue repair/remodeling occurs in both the infarcted and noninfarcted myocardium. Apoptosis has been demonstrated to play an important role in these processes. In the present study, we sought to determine the temporal and spatial characteristics of apoptosis in the infarcted heart as well as to identify cells undergoing programmed cell death at different stages of repair/remodeling and their relationship to the expression of anti-/pro-apoptotic genes following MI. Our study has shown that apoptosis appears in both infarcted and noninfarcted myocardium, and cells undergoing apoptosis depend on the stage of healing. In the infarcted myocardium, apoptosis contributes to the loss of cardiomyocytes during the early stage of healing, elimination of inflammatory cells during the inflammatory phase of healing, and reduction of myofibroblasts with the fibrogenic phase of repair in the infarcted myocardium. In noninfarcted myocardium, cardiomyocyte apoptosis was observed from day 3 to 28 postMI. Cardiac apoptosis following MI is correlated with the increase of Bax expression. PMID- 15530437 TI - Modeling the Trypanosoma cruzi Tc85-11 protein and mapping the laminin-binding site. AB - Trypanosoma cruzi expresses a set of glycoproteins encoded by the gp85/trans sialidase gene superfamily. In this report a structure model is proposed for a cloned member of the superfamily, the Tc85-11 protein. The structure consists of an N-terminus beta-propeller and a C-terminus beta-sandwich interconnected by an alpha-helix. The recombinant protein, corresponding to the N-domain (Tc85-N), binds to laminin in a selective manner. Six synthetic 20-mer peptides from the N domain adhere onto the surface of LLC-MK(2) cells and two of these peptides specifically inhibit the Tc85-N/laminin interaction, indicating that they are the laminin-binding sites of the molecule. Thus, Tc85-11 and other related members of the family appear to be good candidates to play an important role in T. cruzi infection via a laminin mediated host-parasite interaction. PMID- 15530438 TI - Characterization of the bovine neurofilament NF-M protein and cDNA sequence, and identification of in vitro and in vivo calpain cleavage sites. AB - Neurofilaments are the major components of the neuronal cytoskeleton, and accumulations of these proteins are associated with several important human diseases. Here we report the cloning and sequencing of bovine NF-M, the first NF M cloned from a large domestic mammal. The bovine sequence proves to be generally more similar to that of human NF-M than the previously described mammalian sequences, suggesting that bovine neurofilaments are a useful model for biochemical studies of application to humans. However, we noted some unusual features within the 16 lys-ser-pro (KSP2) type phosphopeptide repeats and also note that the number of these repeats correlates well with the size of animal. We also characterized two in vitro calpain cleavage sites by direct peptide sequencing, finding that both are located in the glutamic acid rich E segment. Finally, we show biochemically that the more abundant and stable of these calpain fragments can also be detected in vivo. PMID- 15530439 TI - ATFC is a novel transducer for the unfolded protein response in Bombyx mori BM5 cells. AB - We report the isolation of activating transcription factor of chaperone (ATFC), a novel cDNA from Bombyx mori BM5 cells that encodes a putative transducer of endoplasmic reticulum (ER) stress. The 236 amino acids of ATFC include both a basic region and a leucine zipper at the C-terminus, in contrast to Hac1p of yeast which features these structures at its N-terminus. ATFC expression was strongly up-regulated by ER stress. ATFC could specifically bind to the unfolded protein response element. BM5 cells transfected with ATFC cDNA displayed enhanced ER chaperone expression in response to ER stress. These results indicate that ATFC encodes a putative transducer of ER stress. PMID- 15530441 TI - GEC1, a protein related to GABARAP, interacts with tubulin and GABA(A) receptor. AB - We have previously identified in uterine cells a novel estrogen-regulated gene called gec1. GEC1 presents 87% identity with GABARAP which, so far, was the only protein found to associate with tubulin and GABA(A) receptor. We demonstrated then that GEC1 interacts in vitro with tubulin and GABA(A) receptor, and promotes tubulin assembly and microtubule bundling. Since all polyclonal antibodies failed in discrimination of both proteins GEC1 and GABARAP, a GEC1-GFP fusion protein was used to specifically localize GEC1. GEC1-GFP was distributed over the cytoplasm in perinuclear vesicles with a scattered pattern. Overall, our data show that GEC1 could be a new member of the GABARAP family involved in the transport of GABA(A) receptor. PMID- 15530440 TI - Compounds stimulating cytosolic phospholipase A2 activity with a combinational action mode. AB - The screening of small synthetic compound libraries is a useful means of identifying molecules that modulate various cellular responses. We screened more than 10,000 different small compounds and identified three synthetic compounds that stimulate arachidonic acid (AA) release in a combinational manner in neutrophil-like differentiated HL60 cells. These three compounds were designated as AARIC-1, -2, and -3, representing AA release inducing compounds-1, -2, and -3. Although AA release was not induced by any single one of these compounds, it was dramatically stimulated by the three compounds in combination. Moreover, the effect of combined treatment by these compounds on AA release was completely abolished by MAFP and AACOCF(3), specific cytosolic phospholipase A(2) inhibitors. Furthermore, we found that AARIC-3 stimulates cytosolic calcium influx, while AARIC-1 induces ERK activation. Taken together, we demonstrate a useful approach to the study of complicated and nonlinear intracellular signaling networks using small synthetic compounds in combination. PMID- 15530442 TI - Membrane Biophysics symposium. December 6, 2003, Ann Arbor, Michigan, USA. PMID- 15530443 TI - Direct observation of lipid bilayer disruption by poly(amidoamine) dendrimers. AB - Atomic force microscopy (AFM) is employed to observe the effect of poly(amidoamine) (PAMAM) dendrimers on 1,2-dimyristoyl-sn-glycero-3 phosphocholine (DMPC) lipid bilayers. Aqueous solutions of generation 7 PAMAM dendrimers cause the formation of holes 15-40 nm in diameter in previously intact bilayers. This effect is observed for two different branch end-groups--amine and carboxyl. In contrast, carboxyl-terminated core-shell tectodendrimer clusters do not create holes in the lipid membrane but instead show a strong affinity to adsorb to the edges of existing bilayer defects. A possible mechanism for the formation of holes in the lipid bilayer is proposed. The dendrimers remove lipid molecules from the substrate and form aggregates consisting of a dendrimer surrounded by lipid molecules. Dynamic light scattering (DLS) measurements as well as 31P NMR data support this explanation. The fact that tectodendrimers behave differently suggests that their cluster-like architecture plays an important role in their interaction with the lipid bilayer. PMID- 15530444 TI - Direct AFM observation of saposin C-induced membrane domains in lipid bilayers: from simple to complex lipid mixtures. AB - Saposin C (Sap C) is a small glycoprotein required by glucosylceramidase (GCase) for hydrolysis of glucosylceramide to ceramide and glucose in lysosomes. The molecular mechanism underlying Sap C stimulation of the enzyme activation is not fully understood. Here, atomic force microscopy (AFM) has been used to study Sap C-membrane interactions under physiological conditions. First, to establish how Sap C-membrane interactions affect membrane structure, lipid bilayers containing zwitterionic and anionic phospholipids were used. It was observed that Sap C induced two types of membrane restructuring effects, i.e., the formation of patch like domains and membrane destabilization. Bilayers underwent extensive structural reorganization. To validate the biological importance of the membrane restructuring effects, interaction of Sap C with lipid bilayers composed of cholesterol, sphingomyelin, and zwitterionic and anionic phospholipids were studied. Although similar membrane restructuring effects were observed, Sap C membrane interactions, in this case, were remarkably modulated and their effects were restricted to a limited area. As a result, nanometer-sized domains were formed. The establishment of a model membrane system will allow us to further study the dynamics, structure and mechanism of the Sap C-associated membrane domains and to examine the important role that these domains may play in enzyme activation. PMID- 15530445 TI - Aluminum binding to phosphatidylcholine lipid bilayer membranes: 27Al and 31P NMR spectroscopic studies. AB - 27Al and 31P nuclear magnetic resonance (NMR) spectroscopies were used to investigate aluminum interactions at pH 3.4 with model membranes composed of 1 palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). A solution state 27Al NMR difference assay was developed to quantify aluminum binding to POPC multilamellar vesicles (MLVs). Corresponding one-dimensional (1D) fast magic angle spinning (MAS) 31P NMR spectra showed that aluminum induced the appearance of two new isotropic resonances for POPC shifted to -6.4 ppm and -9.6 ppm upfield relative to, and in slow exchange with, the control resonance at -0.6 ppm. Correlation of the (27)Al and (31)P NMR binding data revealed a 1:2 aluminum:phospholipid stoichiometry in the aluminum-bound complex at -9.6 ppm and a 1:1 aluminum:phospholipid stoichiometry in that at -6.4 ppm. Slow MAS 31P NMR spectra demonstrated shifts in the anisotropic chemical shift tensor components of the aluminum-bound POPC consistent with a close coordination of aluminum with phosphorus. A model of the aluminum-bis-phospholipid complex is proposed on the basis of these findings. PMID- 15530446 TI - Non-raft forming sphingomyelin-cholesterol mixtures. AB - Sphingomyelin from biological membranes forms segregated domains with cholesterol in fluid bilayers. However, a synthetic form of sphingomyelin with an oleoyl chain linked to sphingosine is not incorporated into cholesterol-rich domains. We have studied the properties of mixtures of oleoyl-sphingomyelin and cholesterol as well as mixtures of oleoyl-sphingomyelin with 1-stearoyl-2-oleoyl phosphatidylcholine by DSC and NMR. Cholesterol has a high miscibility with oleoyl-sphingomyelin and it does not separate in crystalline form until the mol fraction of cholesterol reaches a value above 0.6. A large fraction of the cholesterol crystals that are formed are in the monohydrate form. Furthermore, these crystals rehydrate relatively rapidly compared with pure cholesterol crystals in the absence of phospholipid. The environment of the carbonyl group of the phospholipid indicates that it is similar to other forms of sphingomyelin with saturated acyl chains. Also similar to other forms of sphingomyelin, the quaternary ammonium group of oleoyl-sphingomyelin is more rigid than that of phosphatidylcholines, as indicated by the strong resonance observed with cross polarization/magic angle spinning. Additionally, oleoyl-sphingomyelin produces a larger alteration than egg sphingomyelin of the phase transition of 1-stearoyl-2 oleoyl-phosphatidylcholine. These studies indicate that oleoyl-sphingomyelin, unlike saturated forms of sphingomyelin, does not form segregated domains with cholesterol because of its greater miscibility with phosphatidylcholine. PMID- 15530447 TI - Investigation of the interaction of myelin basic protein with phospholipid bilayers using solid-state NMR spectroscopy. AB - Interaction of bovine myelin basic protein and its constituent charge isomers (C1 C3) with phospholipid bilayers was studied using solid-state NMR experiments on model membranes. 31P NMR experiments on multilamellar vesicles and mechanically aligned bilayers were used to measure the degree of protein-induced disorder in the lipid headgroup region while 2H NMR data provided the disorder caused by the protein in the hydrophobic core of the bilayers. Our results suggest that MBP and its charge isomers neither fragment nor significantly disrupt DMPC, POPC, POPC:POPG, and POPE bilayers. These results demonstrate that the MBP-induced fragmentation of POPC bilayers is due to the freeze-thaw cycles used in the preparation of multilamellar vesicles and not due to intrinsic protein-lipid interactions. PMID- 15530448 TI - A 2H solid-state NMR spectroscopic investigation of biomimetic bicelles containing cholesterol and polyunsaturated phosphatidylcholine. AB - Deuterium solid-state NMR spectroscopy was used to qualitatively study the effects of both 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphatidylcholine (PLiPC) and cholesterol on magnetically aligned phospholipid bilayers (bicelles) as a function of temperature utilizing the chain-perdeuterated probe 1,2-dimyristoyl sn-glycero-3-phosphatidylcholine (DMPC-d54) in DMPC/dihexanoylPC (DHPC) phospholipid bilayers. The results demonstrate that polyunsaturated PC and cholesterol were successfully incorporated into DMPC/DHPC phospholipid bilayers, leading to a bicelle that will be useful for investigations of eukaryotic membrane protein-lipid interactions. The data indicate that polyunsaturated PC increases membrane fluidity and decreases the minimum magnetic alignment temperature for DMPC/DHPC bicelles. Conversely, the introduction of cholesterol into aligned DMPC/DHPC bilayers decreases fluidity in the membrane and increases the minimum temperature necessary to magnetically align the phospholipid bilayers. Finally, the addition of Tm3+ to magnetically aligned DMPC/DMPC d54/PLiPC/DHPC bilayers doubles the quadrupolar splittings, indicating that this unique bicelle system can be aligned with the bilayer normal parallel to the static magnetic field. PMID- 15530449 TI - Interaction of the fusogenic peptide B18 in its amyloid-state with lipid membranes studied by solid state NMR. AB - The interaction of the fusogenic polypeptide segment "B18" from the fertilization protein binding with lipid membranes was investigated by solid state 2H and 31P NMR, and by differential scanning calorimetry. B18 is known to adopt different conformations depending on peptide concentration, ionic conditions, pH and lipid environment. Here, the peptide was studied in its beta-stranded amyloid conformation. According to 31P NMR, the lamellar morphology of the DMPC bilayer remains intact in the presence of B18. In going from low (1:90) to high (1:10) peptide/lipid ratios, an increasing effect on several different 2H-labeled lipid segments was observed, reflecting changes in phase behavior and local dynamics. The strongest influence of B18 was detected at the acyl-chains, while no significant effect on the lipid headgroup conformation was observed. This suggests an insertion of B18 in its fibrillar state into the membrane driven by hydrophobic interactions, rather than a peripheral binding mediated by electrostatics. PMID- 15530450 TI - Order from disorder, corralling cholesterol with chaotic lipids. The role of polyunsaturated lipids in membrane raft formation. AB - A myriad of health benefits including the prevention of cancer and heart disease accompanies consumption of polyunsaturated fatty acids (PUFA). Of special importance is the omega-3-PUFA docosahexaenoic acid (DHA), with 22 carbons and six double bonds that constitute the most highly unsaturated fatty acid naturally occurring. Our experiments target the membrane as a likely site of action and focus upon the interaction of cholesterol with PUFA-containing phospholipids. They support the idea that steric incompatibility of the rigid steroid moiety for highly disordered PUFA chains promotes lateral segregation of lipids into PUFA rich/sterol-poor and PUFA-poor/sterol-rich regions. Solid state 2H NMR and X-ray diffraction demonstrate that the solubility of cholesterol is low in polyunsaturated bilayers. In mixed membranes of phosphatidylethanolamine (PE) with the lipid raft-forming molecules sphingomyelin (SM) and cholesterol, diminished affinity of the sterol for 1-[2H31]palmitoyl-2 docosahexaenoylphosphatidylethanolamine ([2H31]16:0-22:6PE) relative to 1 [2H31]palmitoyl-2-oleoylphosphatidylethanolamine ([2H31]16:0-18:1PE) is identified by 2H NMR order parameters. Here, lies the origin of a potential biological advantage of the relatively modest increase in PUFA content of plasma membranes that would be conferred by dietary supplementation. We hypothesize that the enhanced propensity to form SM-/cholesterol-rich rafts as well as PUFA rich/cholesterol-poor microdomains would modify the function of proteins for which these respective regions provide a platform. PMID- 15530451 TI - Solid-state nuclear magnetic resonance studies of HIV and influenza fusion peptide orientations in membrane bilayers using stacked glass plate samples. AB - The human immunodeficiency virus (HIV) and influenza virus fusion peptides are approximately 20-residue sequences which catalyze the fusion of viral and host cell membranes. The orientations of these peptides in lipid bilayers have been probed with 15N solid-state nuclear magnetic resonance (NMR) spectroscopy of samples containing membranes oriented between stacked glass plates. Each of the peptides adopts at least two distinct conformations in membranes (predominantly helical or beta strand) and the conformational distribution is determined in part by the membrane headgroup and cholesterol composition. In the helical conformation, the 15N spectra suggest that the influenza peptide adopts an orientation approximately parallel to the membrane surface while the HIV peptide adopts an orientation closer to the membrane bilayer normal. For the beta strand conformation, there appears to be a broader peptide orientational distribution. Overall, the data suggest that the solid-state NMR experiments can test models which correlate peptide orientation with their fusogenic function. PMID- 15530452 TI - Dynamic pictures of membrane proteins in two-dimensional crystal, lipid bilayer and detergent as revealed by site-directed solid-state 13C NMR. AB - We have compared site-directed 13C solid-state NMR spectra of [3-13C]Ala- and/or [1-13C]Val-labeled membrane proteins, including bacteriorhodopsin (bR), pharaonis phoborhodopin (ppR), its cognate transducer (pHtrII) and Escherichia coli diacylglycerol kinase (DGK), in two-dimensional (2D) crystal, lipid bilayers, and detergent. Restricted fluctuation motions of these membrane proteins due to oligomerization of bR by specific protein-protein interactions in the 2D crystalline lattice or protein complex between ppR and pHtrII provide the most favorable environment to yield well-resolved, fully visible 13C NMR signals for [3-13C]Ala-labeled proteins. In contrast, several signals from such membrane proteins were broadened or lost owing to interference of inherent fluctuation frequencies (10(4)-10(5)Hz) with frequency of either proton decoupling or magic angle spinning, if their 13C NMR spectra were recorded as a monomer in lipid bilayers at ambient temperature. The presence of such protein dynamics is essential for the respective proteins to achieve their own biological functions. Finally, spectral broadening found for bR and DGK in detergents were discussed. PMID- 15530453 TI - Binding and insertion of alpha-helical anti-microbial peptides in POPC bilayers studied by molecular dynamics simulations. AB - We have performed molecular dynamics simulations of the interactions of two alpha helical anti-microbial peptides, magainin2 and its synthetic analog of MSI-78, with palmitoyl-oleoyl-phosphatidylcholine (POPC) lipid bilayers. We used various initial positions and orientations of the peptide with respect to the lipid bilayer, including a surface-bound state parallel to the interface, a trans membrane state, and a partially inserted state. Our 20 ns long simulations show that both magainin2 and MSI-78 are most stable in the lipid environment, with the peptide destabilized to different extents in both aqueous and lipid/water interfacial environments. We found that there are strong specific interactions between the lysine residues of the peptides and the lipid head-group regions. MSI 78, owing to its large number of lysines, shows better binding characteristics and overall stability when compared to magainin2. We also find that both peptides destabilize the bilayer environment, as observed by the increase in lipid tail disorder and the induction of local curvature on the lipid head-groups by the peptides. From all the simulations, we conclude that the hydrogen bonding interactions between the lysines of the peptides and the oxygens of the polar lipid head-groups are the strongest and determine the overall peptide binding characteristics to the lipids. PMID- 15530454 TI - Determination of helical membrane protein topology using residual dipolar couplings and exhaustive search algorithm: application to phospholamban. AB - Dipolar waves are distinct hallmarks of both the secondary and tertiary structures of alpha-helical proteins that are immobilized in membrane bilayers or embedded in anisotropic media. We present a simple, semi-empirical approach that exploits the modulation of the amplitude and average of dipolar waves to determine the topology of alpha-helical proteins. Moreover, we describe the application of this method for the structural determination of a detergent solubilized membrane protein, phospholamban (PLB) that is involved in calcium regulation of cardiac muscle. When combined with high-resolution solid-state NMR data, this method can serve as a fast route for determining the topology of helical membrane proteins solubilized in detergent micelles. PMID- 15530455 TI - Laboratory measurement of growth hormone. AB - Growth hormone (GH) measurements are complicated by the heterogeneous nature of GH, as well as by the presence of the GH binding protein in plasma. Several isoforms of GH exist, and specific assays for each are currently either unavailable, impractical, or not clinically indicated. Bioassays include the in vivo assays based on rat weight gain, tibial line widening, or IGF-I generation. In vitro bioassays, based on the proliferation of cell lines expressing the prolactin receptor or GH receptor, are sensitive but prone to nonspecific interference by factors present in serum. Immunoassays (RIA, IRMA, ELISA, and immunofunctional assay design) are widely used in the clinical laboratory because of speed, sensitivity, and convenience. Discrepancies among results rendered by different immunoassays have become more apparent as monoclonal assays have superseded polyclonal assays, presumably because different antibodies recognize different epitopes among the heterogeneous mixture of GH isoforms in serum. Some assays, especially those with short, nonequilibrium incubation times are vulnerable to interference by the GH binding protein present in serum. Recommendations are given for strategies designed to minimize disparity of results obtained by different GH immunoassays applied to serum. Urinary GH measurements, while technically feasible, are of limited clinical utility because of biological variation in urinary GH excretion. PMID- 15530456 TI - Autoantibodies to citrullinated proteins in rheumatoid arthritis: clinical performance and biochemical aspects of an RA-specific marker. AB - Rheumatoid arthritis (RA) is a common, systemic autoimmune disease of which the exact etiology is not known. In the past 10 years, substantial progress has been made in the identification of the antigens specifically recognized by the autoantibodies of RA patients. A central factor in this respect is citrullination, a form of post-translational modification that is strongly associated with autoimmunity in RA. Here, we summarize and discuss our current knowledge on (i) autoantibody systems in RA, (ii) the occurrence of peptidylarginine deiminases and (iii) citrullinated proteins in natural and diseased environments, and (iv) genetic factors involved in RA that may influence the generation and presentation of citrullinated proteins and the resulting antibody production against these modified proteins. Citrullination of proteins may play a key role in the initiation and/or the progression of RA. The onset of citrulline-specific autoimmunity in RA is probably mediated by both environmental and genetic factors, and future studies will learn whether therapeutic intervention at the level of citrullination may provide new possibilities to treat RA. PMID- 15530457 TI - Protein-to-creatinine ratio in spot urine samples as a predictor of quantitation of proteinuria. AB - BACKGROUND: Normal individuals usually excrete very small amounts of protein in the urine. Persistently increased protein excretion is usually a marker of kidney damage. Quantifying protein in urine is commonly used in the diagnosis of kidney diseases, detection of treatment effects and evaluation of prognosis. We evaluated the use of the total protein-to-creatinine ratio (P/C) in spot urine specimens as a predictor of urine protein excretion in 24-h collections. METHODS: The correlation between P/C in first morning and random urine specimens and urinary protein excretion in 24-h collections were analyzed. The cutoff value of P/C in first morning urine specimens for screening urinary protein excretion of 1 and 3 g in 24-h collections was determined by receiver operating characteristics (ROC) curve. RESULTS: For patients with Ccr 10 ml/min, the correlation was highly significant. Similar results were obtained for random urine specimens. By ROC curve analysis, the P/C of 0.94 and 2.84 g/gcr in first morning urine specimens represent the best threshold to detect urine protein excretion of 1 and 3 g in 24-h collections, respectively. There is a good correlation between P/C in first morning urine specimens and random urine specimens from inpatients and outpatients. But the P/C in random specimens is significantly higher than that in first morning specimens in outpatients. CONCLUSION: The P/C in spot urine samples could be used as an alternative to urine protein excretion in 24-h collections in patients with Ccr>10 ml/min. The P/C in first morning urine samples is better than that in random specimens, especially for outpatients. PMID- 15530458 TI - Tryptophan metabolism along the kynurenine pathway in diet-induced and genetic hypercholesterolemic rabbits. AB - BACKGROUND: The activity of nicotinic acid in hypercholesterolemia has been poorly understood. In man, nicotinic acid derives for the most part from tryptophan along the tryptophan-nicotinic acid pathway, also called the kynurenine pathway, kynurenine being the key metabolite in this process. In the present paper, we investigated if, in animals with hypercolesterolemia, degradation of tryptophan to nicotinic acid along the kynurenine pathway was perturbated. METHODS: Liver, kidney and intestine enzyme activities of the tryptophan-nicotinic acid pathway in normolipidemic, diet-induced hyperlipidemic New Zealand and heritable hypercholesterolemic Watanabe (WHHL) rabbits were determined. RESULTS: Liver tryptophan 2,3-dioxygenase (TDO) activity was present only as a holoenzyme and was higher in the controls than in the hyperlipidemic and Watanabe rabbits, but no difference was present between the group fed an atherogenic hyperlipidic diet and the WHHL rabbits. Small intestine indole 2,3 dioxygenase (IDO) did not vary significantly among the three groups but was higher in comparison with liver TDO activity. In liver, kynurenine 3 monooxygenase and kynurenine-oxoglutarate transaminase activities did not show any significant difference among the three groups of rabbits. Kynureninase and 3 hydroxyanthranilate 3,4-dioxygenase activities per g of fresh tissue decreased significantly in the group of hyperlipidemic and in WHHL rabbits. In the kidneys, kynurenine 3-monooxygenase and kynureninase activity did not change significantly in the three groups of rabbits; kynurenine-oxoglutarate transaminase activity per g of fresh tissue decreased in both hyperlipidemic groups, but no significant difference was observed between hyperlipidemic and Watanabe rabbits. 3 Hydroxyanthranilate 3,4-dioxygenase activity in kidney was decreased markedly in hyperlipidemic and Watanabe rabbits, but there was no difference between the two hypercholesterolemic groups. Aminocarboxymuconate-semialdehyde decarboxylase activity did not change. Thus 3-hydroxyanthranilate 3,4-dioxygenase may be an important regulatory mechanism in the control of the flow of tryptophan along the kynurenine pathway to NAD in hypercholesterolemic rabbits. CONCLUSIONS: This study first demonstrates that in rabbits, hypercholesterolemia, both diet- or genetically induced, can influence the enzyme activities of the tryptophan nicotinic acid pathway leading to a decreased formation of nicotinic acid, and thus NAD. PMID- 15530459 TI - Lack of association of eNOS (G894T) and p22phox NADPH oxidase subunit (C242T) polymorphisms with systemic sclerosis in a cohort of French Caucasian patients. AB - OBJECTIVE: To assess the influence of endothelial nitric oxide synthase (eNOS) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase polymorphisms on the susceptibility of patients to and clinical expression of systemic sclerosis (SSc). METHODS: Seventy-seven French Caucasian patients with SSc were studied. Patients and ethnically matched controls (n=49) were genotyped, by restriction enzyme digestion of polymerase chain reaction (PCR) products, for G894T polymorphism in exon 7 of the eNOS gene and for C242T polymorphism of the gene encoding the p22(phox) NADPH oxidase subunit. RESULTS: The allele and genotype frequencies of the polymorphisms did not differ between patients with SSc and the controls. Moreover, there was no association between these polymorphisms and disease phenotypes. CONCLUSION: Our results indicate that eNOS (G894T) and p22(phox) (C242T) polymorphisms do not influence susceptibility to and the course of systemic sclerosis. PMID- 15530460 TI - Oxidative stress in urogenital tuberculosis patients: a predisposing factor for renal stone formation--amelioration by vitamin E supplementation. AB - BACKGROUND: Previous studies have shown that urogenital tuberculosis (GuTb) patients treated or untreated with regular anti-Tb regimen excrete comparatively high levels of urinary stone forming constituents than normal subjects. Enhanced oxidative stress is also considered as a prime factor that accelerates urolithiasis. The present study was aimed to determine antioxidant status and lipid peroxidation of these individuals in order to assess their risk for kidney stone formation. METHODS: GuTb patients and age-matched normal subjects were divided into four groups: I: normal subjects (n=60), II: GuTb patients a day before treatment (n=72), III: GuTb patients after treatment with isoniazid (300 mg), rifampicin (450 mg) and pyrazinamide (1.5 g) per day for 60 days (n=42), and IV: GuTb patients supplemented with vitamin E (200 mg/day) along with regular chemotherapy for 60 days (n=30). Blood was collected and tested for various markers of oxidative stress. RESULTS: Increased levels of lipid peroxidation, protein carbonyls (PCO), advanced oxidative protein products (AOPP) and reduced antioxidant defenses by impairment in enzyme activities like superoxide dismutase, catalase, glutathione peroxidase, reduced glutathione and decreased plasma concentrations of non enzymatic antioxidants like vitamins C and E were observed in the treated and untreated GuTb patients. CONCLUSIONS: These biochemical disparities may lead to membrane disintegrity, which is favorable for retention of mirolithis. Advocation of vitamin E enhanced the antioxidant status of the plasma, thereby preventing membrane injury, consequently reducing the risk of stone formation in urogenital tuberculosis patients, who were treated with their routine anti-tuberculosis drug regimen. PMID- 15530461 TI - Altered expression of anti and proapoptotic proteins during chemoprevention of hamster buccal pouch carcinogenesis by tomato and garlic combination. AB - BACKGROUND: Effective combinations of dietary agents are promising candidates for cancer chemoprevention because of their safety and the fact that they are not perceived as medicine. The present study was designed to investigate the apoptosis-inducing effect of combined administration of tomato and garlic during 7,12-dimethylbenz[a]anthracene (DMBA)-induced hamster buccal pouch (HBP) carcinogenesis. METHODS: Hamsters were divided into four groups. The right buccal pouches of animals in group 1 were painted with 0.5% DMBA three times a week. Animals in group 2 painted with DMBA as in group 1, received in addition intragastric administration of a combined dose of tomato and garlic on days alternate to DMBA application. Group 3 animals were given chemopreventive agents alone. Animals in group 4 served as control. All the animals were sacrificed after an experimental period of 14 weeks. DNA fragmentation and the apoptosis associated proteins-Bcl-2, Bax, Bim, P53 as well as caspases 8 and 3 were used as markers of apoptosis. RESULTS: Topical application of DMBA for 14 weeks resulted in well-developed squamous cell carcinomas (SCCs) associated with increased expression of Bcl-2 and decreased expression of Bax, Bim, P53 and caspases 8 and 3. Combined administration of tomato and garlic significantly inhibited the development of HBP carcinomas and induced apoptosis. This was evidenced by downregulation of Bcl-2 and upregulation of Bax, Bim, P53 and caspases 8 and 3. CONCLUSION: The induction of apoptosis may be one of the mechanisms through which functional foods such as tomato and garlic exert their anticancer properties. PMID- 15530462 TI - Protective effect of piperine on benzo(a)pyrene-induced lung carcinogenesis in Swiss albino mice. AB - BACKGROUND: The statistics on lung cancer necessitates the urge to develop new methods to control this most deadly form of cancer. Chemoprevention is one of these new approaches. Carcinogens from cigarette smoke form the link between nicotine addiction and lung cancer. At the same time, it has become increasingly clear that dietary and genetically determined factors play an important role in modulating the individual susceptibility and are linked to the chemoprevention approach. In this study, the ability of piperine to prevent lung carcinogenesis in mice was characterized and its effects on cell proliferation and protein damage biomarkers were determined. METHODS: Thirty Swiss albino mice were divided into five groups of six animals each. Animals in group I received corn oil orally and served as control. Group II were administered with 50 mg/kg b.wt. of benzo(a)pyrene (B(a)P) twice a week for 4 consecutive weeks to induce lung cancer by the end of 16th week. Group III received 50 mg/kg b.wt. piperine on alternate days for 16 weeks immediately after the first dose of carcinogen. Group IV also received piperine as in group III but from the sixth week of B(a)P induction till the end of the experiment. Group V animals constituted the drug control and received piperine alone. RESULTS: Administration of piperine significantly decreased the levels of lipid peroxidation, protein carbonyls, nucleic acid content and polyamine synthesis that were found to be increased in lung cancer bearing animals. CONCLUSIONS: Piperine could effectively inhibit B(a)P-induced lung carcinogenesis in albino mice by offering protection from protein damage and also by suppressing cell proliferation. PMID- 15530463 TI - Effects of two P/S ratios with same peroxidizability index value and antioxidants supplementation on serum lipid concentration and hepatic enzyme activities of rats. AB - BACKGROUND: High polyunsaturated/saturated fatty acid (P/S) ratio of diet decreases the risk of cardiovascular disease (CVD) with improvement of serum lipid profiles and inhibition of lipid peroxidation. This study was to investigate the effects of P/S ratio and antioxidants supplementation on lipid and peroxidation when dietary peroxidizability index (PI) value was the same. METHODS: Female Sprague-Dawley rats weighing 100-120 g were fed four different experimental diets for 4 weeks. The P/S ratios were 0.38 (LP) and 4.81 (HP) when the PI value of diet was 81.22. These diets were supplemented with vitamin E 1000 mg/kg diet and selenium 2.5 mg/kg diet (LPS and HPS). RESULTS: The total cholesterol (T-C) and HDL-cholesterol concentrations of serum were higher in HP and HPS (p<0.001). The cardiac index and atherogenic index were lower in HP and HPS (p<0.001). Hepatic thiobarbituric acid reactive substance concentrations were higher in LP and LPS (p<0.001). However, P/S ratio and antioxidants had no effect on antioxidant enzyme activities in the liver. CONCLUSION: A high P/S ratio diet has a beneficial effect on CVD risk even without antioxidant when the PI value is the same. We suggest that P/S ratio as well as PI value should be considered for reducing the risk of CVD. PMID- 15530464 TI - Separation, identification of uremic middle molecules, and preliminary study on their toxicity. AB - BACKGROUND: Compounds accumulating in uremic serum with molecular mass from 300 to 5000 Da are called uremic middle molecules (UMMs). In our previous work, two UMM fractions A and B were obtained from uremic sera, urine, and normal urine by gel permeation chromatography (GPC), and six UMMs from subfraction A3 of uremic plasma and normal urine were purified and characterized. METHODS: Urine and serum samples from uremic patients and healthy subjects were isolated by GPC, ion exchange chromatography (IEC), and reversed-phase high-performance liquid chromatography (RP-HPLC). Moreover, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography/electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) were used to characterize the compounds. The effects of subfraction A3 on renal function were studied in rabbit models with chronic renal failure (CRF). RESULTS: A compound with molecular weight 1007.94 in subfraction A3 was determined to be an octapeptide by mass spectrometry, with an amino acid sequence of Val-Val-Arg Gly-Cys-Thr-Trp-Trp. Two CRF rabbits injected with A3 died in 5 days, while the other two CRF rabbits (no injection) survived a few days. By multistep chromatography and MALDI-TOF MS, another 11 endogenous compounds were found not only in the subfraction B9 of uremic sera but also in that of normal urine. CONCLUSION: Seventeen endogenous middle molecular compounds were found in fractions A and B of uremic plasma and normal urine, among them an octapeptide with M(W) 1007.94 in subfraction A3. Preliminary experimental results on rabbits indicate that subfraction A3 could accelerate the death of rabbits with CRF. PMID- 15530465 TI - Determination of creatinine in urine by tandem mass spectrometry. AB - BACKGROUND: Electrospray tandem mass spectrometry (TMS) is a very powerful tool that enables one to perform high sample throughput analysis. This paper describes a method to determine creatinine in urine by tandem mass spectrometry with direct sample infusion into an ion source. METHODS: Samples (50 microl) were diluted with internal standard (IS) (450 microl of 0.667 mmol/l deuterated creatinine). Diluted samples were introduced into mass spectrometer with no prior pretreatment and after purification on ion-exchange 96-column cartridge. Tandem mass spectrometry analyses were performed in selected reaction monitoring mode. Creatinine and creatinine-d(3) were monitored using precursor and product ion settings (m/z 114 to 86 and m/z 117 to 89, respectively). The time of an analysis was 3.015 min. Both TMS methods were compared mutually and with the results obtained by enzymatic and Jaffe method. RESULTS: Linearity was obtained in the range 0.06-60 mmol/l. Detection limit was 0.2 mumol/l and recoveries were in the range 95.1-98.3% for both the assays with and without ion-exchange column. Results of both assays are in good agreement with those obtained by enzymatic and Jaffe method based on log-transformed Bland-Altman plots. Electrospray tandem mass spectrometry method utilizing both approaches with and without ion-exchange column is acceptable according to CLIA criteria. CONCLUSION: Tandem mass spectrometry allows rapid, sensitive and selective determination of creatinine in untreated urine. PMID- 15530466 TI - LCAT-dependent conversion rate is a determinant of plasma prebeta1-HDL concentration in healthy Japanese. AB - BACKGROUND: Prebeta1-HDL acts as a primary acceptor of cellular cholesterol. Prebeta1-HDL is converted into alpha-migrating high-density lipoprotein (HDL) by lecithin/cholesterol acyltransferase (LCAT). We examined whether the LCAT dependent conversion rate of prebeta1-HDL is a determinant of the plasma prebeta1 HDL concentration in healthy Japanese. METHODS: We measured the conversion half time (CHT(prebeta1)), the time required for 50% of baseline prebeta1-HDL to be changed into alpha-migrating HDL by LCAT, in 100 healthy Japanese (47 men, 53 women, 22-88 years). RESULTS: Prebeta1-HDL concentration, as determined by immunoassay, was significantly lower in younger women (<50 years, n=24) than in older women (>or=50 years, n=29) (16.8+/-3.3 vs. 21.7+/-8.0 mg/l apolipoprotein AI (apoAI), p<0.01). There was no significant difference in prebeta1-HDL concentration between younger (n=24) and older (n=23) men (21.2+/-6.8 vs. 22.5+/ 6.6 mg/l apoAI). The mean CHT(prebeta1) for all subjects was 47.4+/-13.0 min, and was not influenced by gender or age. Prebeta1-HDL concentration was positively correlated with CHT(prebeta1) in both men and women, suggesting that high prebeta1-HDL levels may reflect delayed conversion of prebeta1-HDL. CONCLUSION: LCAT-dependent conversion rate is a determinant of plasma prebeta1-HDL concentration in healthy Japanese. We speculate that prebeta1-HDL concentration may be used as a metabolic marker for HDL maturation. PMID- 15530467 TI - Time course of inflammatory response after renal artery stenting in patients with atherosclerotic renal stenosis. AB - BACKGROUND: Inflammatory response has been demonstrated in patients with coronary artery disease after percutaneous coronary intervention (PCI). Such response following renal artery stenting has not yet been established, however, in patients with atherosclerotic renal artery stenosis. METHODS: A total of 44 patients were enrolled in this study. Of them, 22 patients with atherosclerotic renal artery stenosis received renal angioplasty with stent (group A, mean age 51+/-8 years), and 22 patients with age- and gender-matched underwent renal angiography for diagnostic purpose as a control group (group B, age 50+/-8 years). The peripheral blood samples were taken immediately before the procedure, 1, 6 and 24 h after the procedure in both groups. The concentrations of C reactive protein (CRP) and interleukin-6 (IL-6) were measured using ELISA. RESULTS: The result showed that there was no difference in clinical characteristics and baseline levels of CRP and IL-6 between the groups. The IL-6 increased in the first hour (before: 5.8+/-3 pg/ml; 1 h: 8.6+/-5 pg/ml, p<0.01), lasted at 6 h (12.2+/-8 pg/ml), returned to baseline at 24 h (5.4+/-3 pg/ml) in group A. The CRP did not changed at the first hour after stenting, but mean CRP increased from 0.30+/-0.09 to 0.37+/-0.15 mg/dl at 6 h (p<0.05), and peaked at 24 h (0.43+/-0.18 mg/dl, p<0.001 compared with baseline and control) after stenting in group A, while no such changes were observed in group B (p>0.05 at different time points compared with baseline and group B, respectively). CONCLUSIONS: The data indicated that renal artery stenting could trigger inflammatory response by evidence of increased plasma levels of CRP and IL-6. IL-6, however, was an early initiator of inflammatory cytokine, and CRP was a later marker of systemic inflammatory response to renal artery stenting. PMID- 15530468 TI - Effects of beta thalassemia minor on results of six glycated hemoglobin methods. AB - BACKGROUND: Beta-thalassemia minor (BTM) is a common benign condition that can be present in patients with diabetes mellitus. There are conflicting reports about the effect of BTM on glycated hemoglobin (gHb) measurements. We evaluated 6 gHb methods using samples from non-diabetic subjects with BTM. METHODS: Samples submitted for hemoglobin phenotype analysis were evaluated. A total of 57 samples (30 controls and 27 with BTM) from non-diabetic subjects were selected. GHb analysis was performed by Tosoh A1c 2.2+, Primus CLC 330, Bayer DCA 2000, Beckman Coulter, Synchron CX7 and LX20, and Roche Tina-quant II assays. RESULTS: The A1c 2.2+, CLC 330, DCA 2000 and Tina-quant II assays showed no statistically significant difference between the control and BTM groups. In contrast, BTM results were significantly higher than controls on the Synchron CX7 analyzer and borderline significant on the Synchron LX20 (p=0.051). Further investigation demonstrated an increase in Synchron %HbA(1c) results with decreasing hemoglobin concentrations. CONCLUSIONS: In this study using samples from subjects with normal or near-normal gHb, BTM does not affect gHb measurements per se. However, the Synchron methods yielded higher results for samples with lower hemoglobin concentrations, like those that can be seen in BTM. The Synchron method was improved at the end of 2003, which minimized this problem. PMID- 15530469 TI - Transferrin neutralization of amyloid beta 25-35 cytotoxicity. AB - BACKGROUND: Fibrillar aggregates of amyloid beta 25-35 (Abeta(25-35)) form rapidly in vitro able to lyse human red blood cells (RBCs). Human sera, albumin, and apolipoprotein E (ApoE) each limit fibrillation and cytotoxicity. Potentially, these substances protect neurons from Abeta(1-40/42) aggregates. Transferrin (TF) is investigated in this study. METHODS: The Mattson red blood cells model was employed to determine whether co-incubation of transferrin and Abeta(25-35) prevented lysis. The formation of fibrillar Abeta(25-35) in the presence of transferrin was investigated using Congo red staining and spectrophotometric studies. RESULTS: We found that incubation of 20 muM Abeta(25 35) with physiologic levels of transferrin prevented red blood cells lysis and the formation of macro-aggregates. CONCLUSIONS: These in vitro results suggest that transferrin may limit fibrillar beta amyloid formation in vivo and cytotoxicity. PMID- 15530470 TI - Prenatal diagnosis of sickle cell anemia in twin pregnancies and identification by VNTRs. AB - BACKGROUND: Sickle cell anemia (SCA) is an inherited disorder of hemoglobin synthesis that is characterized by life-long severe hemolytic anemia, attacks of pain crisis, and chronic organ system damage. In this study, prenatal diagnosis was performed to three couples (families A, B, and C) with twin pregnancies who were at risk for SCA. METHODS: The SCA carrier state of the couples were confirmed at molecular level. Chorionic villus samples (CVS) of twins were obtained at 10-12 weeks of gestation. Amplification refractory mutation system (ARMS) and restriction fragment length polymorphism (RFLP) techniques were applied to determine and confirm the presence of the sickle cell anemia of the fetuses. Identification of each twin, confirmation of parentage and elimination of maternal contamination of chorionic villus samples were ruled out by variable number of tandem repeats (VNTR) analysis of four different loci [D1S80 (pMCT118), ApoB, IgJH, D4S95]. RESULTS: We found that one of the fetuses was heterozygous for SCA and the other was normal in family A; in family B both fetuses were heterozygous and in family C both fetuses were normal. CONCLUSION: Prenatal diagnosis is the major way of prevention of the genetic disorders including sickle cell anemia. Prenatal diagnosis of twins contains additional risks such as confusion in differentiation leading to false molecular diagnosis. VNTR loci analysis is a useful tool and can be safely used for the elimination of all problems mentioned above. PMID- 15530471 TI - EDTA is essential to recover lead from dried blood spots on filter paper. AB - BACKGROUND: Residual dried blood spots (DBSs) on filter paper from neonatal screening have been proposed as samples for population survey of lead contamination. We have investigated the EDTA effect on lead release in the eluting solution. METHODS: Furnace atomic absorption spectrophotometry has been used for lead measurements. Standard, blank and sample solutions contained 2% m/v NH(4)H(2)PO(4), 0.5% v/v Triton X-100 and 0.2% v/v HNO(3) as matrix modifier solution (MMS) with or without EDTA. A calibration curve was established from aqueous standard solutions. Paper discs from DBS and blank, punched near the DBS, were eluted in MM solution and, where required, EDTA at different concentrations. Specimens were leftover DBSs with different storage times, matched samples from 20 adult patients consisting of liquid whole blood (LWB) containing 5 mmol/L EDTA, DBSs eluted in MM solution with 5 mmol/L EDTA or without EDTA. RESULTS: Optimal lead recovery from DBS required 5 mM EDTA in the eluting solution. Mean lead levels of LWB and DBSs eluted with EDTA were similar and higher than DBSs without EDTA (P<0.001). Without EDTA, the median value of lead optical density was lower for 6-month-old DBSs than for blanks (P<0.001). CONCLUSIONS: Residual DBSs can be used for population survey, but 5 mmol/L EDTA in the extracting solution is required to fully recover lead. PMID- 15530472 TI - Nilvadipine inhibits nuclear factor-kappaB-dependent transcription in hepatic cells. AB - BACKGROUND: Recent findings suggest that some dihydropyridine-type calcium channel blockers, widely used as anti-hypertensive drugs, have direct anti atherogenic action through their antioxidant properties. METHODS: We examined the effect of nilvadipine on the activity of a representative radical-sensitive transcription factor, nuclear factor kappa-B (NF-kappaB), in the human hepatocyte cell line HuH7 in vitro. RESULTS: Nilvadipine potently inhibited NF-kappaB dependent transcription in a dose- and time-dependent manner, with a minimal effective concentration of 50 nmol/l. The effect was specific because no similar effects were found in the prototype dihydropyridine nifedipine. Electromobility shift assay showed reduced protein binding to the NF-kappaB-consensus sequence in nilvadipine-treated cells. Nilvadipine also reduced the expression of fibrinogen and plasminogen activator inhibitor-1 (PAI-1). CONCLUSIONS: Since NF-kappaB mediated gene products, such as fibrinogen and PAI-1, are known to facilitate hypercoagulation, thrombosis and vascular events, we suggest that nilvadipine has a direct beneficial effect separate from its anti-hypertensive properties by inhibiting NF-kappaB-dependent gene expression and eventually inhibiting atherosclerosis. PMID- 15530473 TI - Significance of apolipoprotein(a) phenotypes in acute coronary syndromes: relation with clinical presentation. AB - BACKGROUND: High lipoprotein(a) [Lp(a)] levels and small-sized apolipoprotein(a) [apo(a)] phenotypes have been linked to acute coronary syndromes (ACS). We sought to determine whether Lp(a) concentrations and apo(a) phenotypes may be related to the clinical syndrome of presentation among ACS patients. METHODS: Two hundred ten ACS patients and 105 controls were enrolled. One hundred thirteen patients presented with acute myocardial infarction (AMI) and 97 with unstable angina pectoris (UAP). Lp(a) concentrations were determined by ELISA and apo(a) isoforms were detected with a high-resolution immunoblotting method. RESULTS: Lp(a) levels and the percentage of subjects with at least one small-sized apo(a) isoform were significantly higher both in AMI patients and in UAP subjects as compared with controls. Among ACS patients, the percentage of subjects with at least one small apo(a) phenotype was significantly higher in patients who presented with AMI than in those with UAP (p<0.001). Multivariate logistic regression analysis showed that the presence of at least one small-sized apo(a) isoform was associated with AMI as the patient's clinical syndrome of presentation (OR=2.51, 95% CI: 1.38 4.58, p<0.01). CONCLUSIONS: Among ACS patients, apo(a) isoforms of low molecular weight were associated with AMI onset. High-resolution apo(a) phenotyping might be helpful to identify individuals at high risk for developing AMI. PMID- 15530474 TI - Serum and plasma fragments of C-telopeptides of type I collagen (CTX) are stable during storage at low temperatures for 3 years. AB - BACKGROUND: Control of pre-analytical variables is essential for successful application of biological markers, including bone resorption markers, in clinical trials and routine use. The effect of storage temperature on stability of bone resorption markers have not been subject of systematically investigation, and therefore the present study was set out to determine the stability of C telopeptides of type I collagen (CTX) in serum and plasma samples stored frozen for 3 years. METHODS: The serum and plasma levels of CTX were determined in samples aliquoted and stored frozen for up to 3 years. RESULTS: No significant decrease could be detected in neither serum nor plasma samples after 3 years of storage at -20, -80 or -150 degrees C. However, at elevated temperature, i.e. 4 and 37 degrees C, improved stability of CTX was observed in EDTA plasma samples compared to serum. CONCLUSIONS: CTX is stable in frozen serum and plasma samples for up to 3 years. EDTA plasma might be the preferred matrix due to improved stability at elevated temperatures. PMID- 15530475 TI - Serum amylase, pancreatic amylase and lipase concentrations in epileptic children treated with carbamazepine monotherapy. AB - BACKGROUND: Serum total amylase and lipase activities have been determined in epileptic patients treated with polytherapy using enzyme-inducing anticonvulsant drugs; however, to our knowledge, serum total amylase, pancreatic amylase and lipase activities have not previously been determined in patients receiving carbamazepine monotherapy. The purpose of this study was to investigate by a prospective, self-controlled method, whether early treatment with carbamazepine monotherapy may alter serum total amylase, pancreatic amylase and lipase concentrations of epileptic children. METHODS: Serum total amylase, pancreatic amylase and lipase activities have been determined in 18 epileptic children before and at 6 and 12 months of treatment with carbamazepine monotherapy. Serum gamma-glutamyltransferase activities were also determined. RESULTS: Serum total amylase concentrations were significantly increased at 6 months of treatment (p=0.034), and serum nonpancreatic amylase concentrations were significantly increased at 6 (p=0.016) and 12 months of treatment (p=0.039), whereas serum pancreatic amylase and lipase concentrations did not significantly change at 6 or 12 months of treatment with carbamazepine monotherapy. Furthermore, serum gamma glutamyltransferase concentrations were significantly increased at 6 (p=0.000) and 12 months of treatment (p=0.000) with carbamazepine monotherapy. There was no significant correlation between serum nonpancreatic amylase concentrations and serum gamma-glutamyltransferase or carbamazepine concentrations at 6 and 12 months of treatment with carbamazepine monotherapy. CONCLUSIONS: These findings indicate that nonpancreatic amylase concentrations may be increased in patients treated with carbamazepine monotherapy. Therefore, measurement of serum pancreatic amylase and lipase concentrations is suggested in epileptic patients receiving carbamazepine monotherapy with symptoms suggesting pancreatic dysfunction, so that unnecessary discontinuing of treatment with carbamazepine should be avoided. PMID- 15530476 TI - Hypermethylation of the CpG islands in the promoter region of the GSTP1 gene in prostate cancer: a useful diagnostic and prognostic marker? AB - BACKGROUND: Recently, many studies have focused on the potential diagnostic value of the promoter hypermethylation of the GSTP1 gene in prostate cancer. METHOD: A total of 144 patients, undergoing eight-core prostatic biopsies for a clinically suspected prostate cancer, was analyzed. Two different tissue samples were collected from the same area of the prostate and then divided for both genomic DNA extraction and pathological examination. In order to perform molecular analysis, prostatic tissue samples were digested with the methylation-sensitive restriction enzyme HpaII and then amplified by conventional polymerase chain reaction (PCR). RESULTS: Prostate cancer was diagnosed in 42/144 patients, and promoter hypermethylation of GSTP1 gene was detected in 31/42 of prostate cancer (sensitivity=74%) and in 2/102 of negative specimens (specificity=98%). A significant association between GSTP1 promoter hypermethylation both with a Gleason score >or=7 (Fisher's exact P=0.01) and the presence of Gleason grade 4 and/or grade 5 (Fisher's exact P=0.03) was found. CONCLUSION: Promoter hypermethylation of the GSTP1 gene is a highly specific--but not a very sensitive -marker of prostate cancer. Our data showed a significant association between the methylation status of the GSTP1 gene and Gleason score and grade, suggesting a potential prognostic value of this epigenetic DNA alteration. PMID- 15530477 TI - Potassium flux of erythrocytes in chronic hemodialysis patients. AB - BACKGROUND: In chronic hemodialysis patients, hyperkalemia is frequently observed. In these patients, erythrocytes were examined to know whether they participate in the regulation of K(+) or not. METHODS: Erythrocyte K(+) release (DeltaKr) was induced by the incubation of erythrocyte suspension at 4 degrees C for 24 h and the K(+) influx followed at 37 degrees C for 3 h. K(+) flux of erythrocytes or DeltaKi/DeltaKr ratio, which was reflected by Na(+)/K(+) exchanging ATPase, was measured in chronic hemodialysis patients. K(+) concentration was measured by ion-selective electrode method. RESULTS: Non diabetic hemodialysis patients classified into three groups according to their serum levels were compared for various factors. Among them, the DeltaKi/DeltaKr ratios in medium- and high-serum K(+) groups were significantly lower than those in the low serum K(+) group. The effect of hemodialysis on erythrocyte K(+) flux was examined. After hemodialysis, the mean DeltaKi/DeltaKr ratio increased significantly compared with that before the treatment. Erythrocyte K(+) concentrations converted into a narrower range after hemodialysis. CONCLUSION: The reduced K(+) flux in erythrocyte may play a part in the development of hyperkalemia in non-diabetic chronic hemodialysis patients. PMID- 15530478 TI - Association of interleukin-8 with inflammatory and innate immune components in bronchoalveolar lavage of children with chronic respiratory diseases. AB - BACKGROUND: The host inflammatory response and innate immunity play a complex role in respiratory diseases. METHODS: We evaluated the levels of inflammatory mediators and antibacterial proteins in children who required bronchoscopy and bronchoalveolar lavage fluid (BALF) for clinical indications such as chronic tracheostomy (n=15) and chronic suppurative lung disease (n=8). RESULTS: Our results suggested the presence of interleukin-1beta (IL-1beta) and IL-8 as major inflammatory mediators in BALF samples. The level of the antibacterial protein sIgA was higher than lactoferrin and lysozyme. BALF IL-8 levels significantly correlated with the presence of IL-1beta, IL-6, IL-10, IL-16, sIgA and lysozyme. BALF IL-8 levels did not correlate with the levels of immunomodulatory and anti inflammatory clara cell 10 kDa protein (CC10) or lactoferrin. CONCLUSION: This study suggests that patients with high levels of BALF IL-8 could potentially have high levels of IL-6, IL-10, IL-16, lysozyme and sIgA. Evaluating the inflammatory mediators (IL-8) in relation to other BALF protein components provides insight into understanding the role of inflammatory mediators in the regulation of host defense and the response to lung inflammation and injury. PMID- 15530479 TI - Evaluation of an assay for serum 1,5-anhydroglucitol (GlycoMark) and determination of reference intervals on the Hitachi 917 analyzer. AB - BACKGROUND: 1,5-Anhydroglucitol (1,5-AG) is a glucose analogue, which is decreased in hyperglycemic individuals. We report the technical performance of an assay (GlycoMark) on a chemistry analyzer, evaluation of analyte stability and determination of reference intervals for 1,5-AG in a non-diabetic US population. METHODS: NCCLS protocols were followed to evaluate the reagent on a Hitachi 917 chemistry analyzer. RESULTS: Intra- and interassay imprecision ranged from 1.3% to 3.8% and 0.79% to 3.7%, respectively. The assay was linear to 110 microg/ml. Interference from triglyceride, hemoglobin and bilirubin was <10% to concentrations of 12.6 mmol/l, 12.1 and 911.4 micromol/l, respectively. Correlation coefficients between lot numbers on the Hitachi 917 and between analyses on the Hitachi 917 and the Hitachi 7170 analyzers were >0.99. The lowest limit of detection was 0.49 microg/ml (mean+/-2 S.D.). 1,5-AG was stable at 4 degrees C for 7 days, at 22 degrees C for 5 days, at -80 degrees C for 14 days and for three freeze-thaw cycles at -80 degrees C. The US reference intervals (nonparametric 2.5th-97.5th percentiles) were 10.2-33.8 microg/ml (males) and 5.9 31.8 microg/ml (females). CONCLUSIONS: The performance of the GlycoMark assay for the measurement of 1,5-AG was acceptable on the Hitachi 917 analyzer. PMID- 15530480 TI - Effects of amino acids and its metabolites on prolidase activity against various iminodipeptides in erythrocytes from normal human and a patient with prolidase deficiency. AB - BACKGROUND: The characteristics of prolidase in erythrocytes from controls and patient with prolidase deficiency were investigated. METHODS: The erythrocytes were isolated from the heparinized blood of normal human and a patient with prolidase deficiency. Effects of various amino acids and their metabolites on prolidase activity against iminodipeptides in presence of 1 mmol/l MnCl(2) were investigated. RESULTS: Prolidase activity against glycylproline in erythrocytes from normal human was strongly enhanced by glycine, L-alanine, L-serine with MnCl(2), but the activity was strongly inhibited by L-valine, and L-leucine. However, the stereoisomers, D-leucine and D-valine enhanced the activity. The prolidase activity against methionylproline in erythrocytes from the patient with prolidase deficiency was also enhanced by glycine, L-alanine and L-serine. The activity was inhibited by l-leucine, but D-leucine and L-valine enhanced the activity against various iminodipeptides. CONCLUSION: Prolidase activity against glycylproline in normal human erythrocytes and against methionylproline from the prolidase-deficient patient was enhanced strongly by glycine, alanine and serine with MnCl(2). However, this activity was inhibited by L-leucine, but was enhanced by D-leucine. PMID- 15530481 TI - Liquid chromatography-tandem mass spectrometry method for the simultaneous determination of delta-ALA, tyrosine and creatinine in biological fluids. AB - BACKGROUND: Several acquired and congenital human disorders perturb the concentrations of delta-aminolevulinate (delta-ALA), creatinine and tyrosine in biological fluids. There is currently no facile, sensitive and specific method to measure these analytes simultaneously. METHOD: We developed an LC-MS/MS method to quantify delta-ALA, creatinine and tyrosine in urine that requires minimal sample preparation and no derivatization. The method is also applicable to the analysis of tyrosine in plasma. RESULTS: All calibration plots were linear, with R(2)>or=0.996. Intra- and interday CVs were <10%. The limit of quantitation for delta-ALA was approximately 0.1 micromol/l, and for creatinine and tyrosine it was well below the lowest measured physiological concentrations. The method was applied to analyze urine from 75 healthy volunteers and 43 patients with hereditary tyrosinemia type I (HT I). The mean urinary concentration of delta-ALA in patients (38+/-35 micromol/l, 53+/-30 mg/g creatinine) was higher than that measured in healthy subjects (5.5+/-2.6 micromol/l, 0.9+/-0.2 mg/g creatinine; p<0.001). Treatment with 2-(2-nitro-4-trifluoromethylbenzyl)-1,3-cyclohexanedione (NTBC), an inhibitor of an early step in tyrosine catabolism, decreased urinary delta-ALA (6.4+/-4.8 micromol/l, 13+/-24 mg/g creatinine; p<0.001). The average plasma tyrosine concentration in healthy volunteers (56+/-14 micromol/l) was within normal reference interval used in clinical practice. CONCLUSIONS: The method is simple, specific and precise and allows simultaneous quantitation of delta-ALA, creatinine and tyrosine at concentrations present under physiological or pathophysiological conditions. PMID- 15530482 TI - Effects of temperature on pre-analytical stability of S100B protein concentrations in urine of healthy full-term infants. PMID- 15530483 TI - Measurement of urinary hydrogen peroxide by FOX-1 method in conjunction with catalase in diabetes mellitus--a sensitive and specific approach. PMID- 15530484 TI - Serial serum procalcitonin changes in the prognosis of acute stroke. PMID- 15530485 TI - C-reactive protein concentrations are lower in Singaporeans: implications for risk classification in Asians. PMID- 15530486 TI - Transcriptional frequency and cell determination. AB - The relative base composition of DNA regulatory sequences of certain genes of undetermined multipotent progenitor cells may account for the frequency of transcription of these genes in cell determination. The sequences of these regulatory regions of cell determination genes that are more AT-rich would create the potential for transcription at a higher frequency due to their lower melting temperature, as well as propensity to bend. An increase of one or more of the high mobility group (HMG) chromatin proteins would preferentially bind the more AT-rich regulatory sequences, thereby increasing the rate of transcription. The amount of unphosphorylated H1 histone reacting with these same regulatory sites may decrease transcription frequency. The level of cell growth, i.e. total protein synthesis of a cell, is correlated positively with the synthesis of HMG proteins. H1 histone synthesis is linked to DNA replication. Unbalanced growth would alter the amounts of HMG proteins and H1 histone, thus changing transcriptional frequency. The greater the enrichment of AT sequences in the regulatory regions of the cell determination genes, the greater may be the extent of evolutionary conservation. Higher frequency of transcription of the cell determination genes with the more AT-rich regulatory sequences could account for the earlier expression of the more conserved cell determination genes during embryonic development. Preferential binding of H1 histone to the more AT-rich regulatory sequences would subsequently restrict their transcription before that of less conserved cell determination genes. PMID- 15530487 TI - A computational method for inferring growth parameters and shape changes during development based on clonal analysis. AB - We describe a method for estimating growth parameters in various regions of a developing organ undergoing cell divisions, along with the corresponding changes in organ shape. Growth parameters are computed by coupling clonal analysis with a growth model, allowing a wide range of developmental stages to be covered. The method was applied to the development of dorsal petal lobes of Antirrhinum majus. The resulting description of growth patterns and shape changes is consistent with direct observations using scanning electron microscopy. This method can potentially be applied to other organs, and opens the way to comparative studies of growth and gene expression patterns. PMID- 15530488 TI - Evaluation of pathways for progression of heterogeneous breast tumors. AB - To better understand the progression of heterogeneous breast cancers, four models of progession pathways have been evaluated. The models describe the progression through the grades of ductal carcinoma in situ (DCIS) 1, 2, and 3, and through the grades of invasive ductal carcinoma (IDC) 1, 2, and 3. The first three pathways, termed linear, nonlinear, and branched, describe DCIS as a progenitor of IDC, and grades of DCIS progressing into grades of IDC. The fourth pathway, termed parallel, describes DCIS and IDC as diverging from a common progenitor and progressing through grades in parallel. The best transition rates for the linear, nonlinear, and branched pathways were sought using a random search in combination with a directed search based on the Nelder-Mead simplex method. Parameter values for the parallel pathway were determined with heuristic graphs. Results of computer simulation were compared with clinically observed frequencies of grades of DCIS and grades of IDC that were reported to occur together in heterogeneous tumors. Each of the four pathways could simulate frequencies that resembled, to varying degrees, the clinical observations. The parallel pathway produced the best correspondence with clinical observations. These results quantify the traditional descriptions in which grades of DCIS are the progenitors of grades of IDC. The results also raise the alternative possibility that, in some tumors with both components, DCIS and IDC may have diverged from a common progenitor. PMID- 15530489 TI - Effects of information and group structure on evolution of altruism: analysis of two-score model by covariance and contextual analyses. AB - An altruistic individual has to gamble on cooperation to a stranger because it does not know whether the stranger is trustworthy before direct interaction. Nowak and Sigmund (Nature 393 (1998a) 573; J. Theor. Biol. 194 (1998b) 561) presented a new theoretical framework of indirect reciprocal altruism by image scoring game where all individuals are informed about a partner's behavior from its image score without direct interaction. Interestingly, in a simplified version of the image scoring game, the evolutionarily stability condition for altruism became a similar form of Hamilton's rule, i.e. inequality that the probability of getting correct information is more than the ratio of cost to benefit. Since the Hamilton's rule was derived by evolutionarily stable analysis, the evolutionary meaning of the probability of getting correct information has not been clearly examined in terms of kin and group selection. In this study, we applied covariance analysis to the two-score model for deriving the Hamilton's rule. We confirmed that the probability of getting correct information was proportional to the bias of altruistic interactions caused by using information about a partner's image score. The Hamilton's rule was dependent on the number of game bouts even though the information reduced the risk of cooperation to selfish one at the first encounter. In addition, we incorporated group structure to the two-score model to examine whether the probability of getting correct information affect selection for altruism by group selection. We calculated a Hamilton's rule of group selection by contextual analysis. Group selection is very effective when either the probability of getting correct information or that of future interaction, or both are low. The two Hamilton's rules derived by covariance and contextual analyses demonstrated the effects of information and group structure on the evolution of altruism. We inferred that information about a partner's behavior and group structure can produce flexible pathways for the evolution of altruism. PMID- 15530490 TI - Extinction risk of a meta-population: aggregation approach. AB - Aggregation of variables of a complex mathematical model with realistic structure gives a simplified model which is more suitable than the original one when the amount of data for parameter estimation is limited. Here we explore use of a formula derived for a single unstructured population (canonical model) in predicting the extinction time for a population living in multiple habitats. In particular we focus multiple populations each following logistic growth with demographic and environmental stochasticities, and examine how the mean extinction time depends on the migration and environmental correlation. When migration rate and/or environmental correlation are very large or very small, we may express the mean extinction time exactly using the formula with properly modified parameters. When parameters are of intermediate magnitude, we generate a Monte Carlo time series of the population size for the realistic structured model, estimate the "effective parameters" by fitting the time series to the canonical model, and then calculate the mean extinction time using the formula for a single population. The mean extinction time predicted by the formula was close to those obtained from direct computer simulation of structured models. We conclude that the formula for an unstructured single-population model has good approximation capability and can be applicable in estimating the extinction risk of the structured meta-population model for a limited data set. PMID- 15530491 TI - Membrane transporters as machines: degenerate singularities as a requirement for function. AB - It is suggested that Membrane Transporter functionality is based on low energy paths between proteins of different conformations. A simple extension of the Born Oppenheimer approximation is used to reduce the protein structure problem to one of the kinematics of engineering mechanisms. Such low energy paths between conformations with the same handedness imply the existence of degenerate singularities in the engineering mechanism. The requirement for degeneracy leads to a number of conjectures. These include the structure and function of chaperones for constructing such proteins and the thermodynamic properties of membrane transporters. PMID- 15530492 TI - A simple cellular automaton model for influenza A viral infections. AB - Viral kinetics have been extensively studied in the past through the use of spatially homogeneous ordinary differential equations describing the time evolution of the diseased state. However, spatial characteristics such as localized populations of dead cells might adversely affect the spread of infection, similar to the manner in which a counter-fire can stop a forest fire from spreading. In order to investigate the influence of spatial heterogeneities on viral spread, a simple 2-D cellular automaton (CA) model of a viral infection has been developed. In this initial phase of the investigation, the CA model is validated against clinical immunological data for uncomplicated influenza A infections. Our results will be shown and discussed. PMID- 15530493 TI - Amplification and oscillations in the FAK/Src kinase system during integrin signaling. AB - Integrin signaling is a major pathway of cell adhesion to extracellular matrices that regulates many physiological cell behaviors such as cell proliferation, migration or differentiation and is implied in pathologies such as tumor invasion. In this paper, we focused on the molecular system formed by the two kinases FAK (focal adhesion kinase) and Src, which undergo auto- and co activation during early steps of integrin signaling. The system is modelled using classical kinetic equations and yields a set of three nonlinear ordinary differential equations describing the dynamics of the different phosphorylation forms of FAK. Analytical and numerical analysis of these equations show that this system may in certain cases amplify incoming signals from the integrins. A quantitative condition is obtained, which indicates that the total FAK charge in the system acts as a critical mass that must be exceeded for amplification to be effective. Furthermore, we show that when FAK activity is lower than Src activity, spontaneous oscillations of FAK phosphorylation forms may appear. The oscillatory behavior is studied using bifurcation and stability diagrams. We finally discuss the significance of this behavior with respect to recent experimental results evidencing FAK dynamics. PMID- 15530494 TI - First phase microcirculatory reaction to mechanical skin irritation: a three layer model of a compliant vascular tree. AB - Mechanical skin irritation creates vasodilatation in the line of a stroke and in the surrounding tissue. To obtain further insight on underlying physiological mechanisms we developed a model of the vascular network comprised of three layers, where the first and the last one have a tree structure. They represent the arterial and the venous system, respectively. Both are connected via an intermediate zone representing the core of the microcirculation, which is described by means of a compliant compartment model. Irritation induces change in compliance of vessels situated at the entrance of the intermediate zone. Thus the model describes flow and pressure behavior due to mechanical skin irritation. PMID- 15530495 TI - Time-dependent animal conflicts: 1. The symmetric case. AB - Animal conflicts are often characterized by time-dependent strategy sets. This paper considers the following type of animal conflicts: a member of a group is at risk and needs the assistance of another member to be saved. As long as assistance is not provided, the individual which is at risk has a positive, time dependent rate of dying. Each of the other group members is a potential helper. Assisting this individual accrues a cost, but losing him decreases the inclusive fitness of each group member. A potential helper's interval between the moment an individual finds itself at risk and the moment it assists is a random variable, hence its strategy is to choose the probability distribution for this random variable. Assuming that each of the potential helpers knows the others' strategies, we show that the ability to observe their realizations influences the evolutionarily stable strategies (ESS) of the game. According to our results, where the realizations can be observed ESS always exist: immediate assistance, no assistance and delayed assistance. Where the realizations cannot be observed ESS do not always exist, immediate assistance and no assistance are possible ESS, while delayed assistance cannot be an ESS. We apply our model to the n brothers' problem and to the parental investment conflict. PMID- 15530496 TI - Time-dependent animal conflicts: 2. The asymmetric case. AB - This paper presents an asymmetric game-theoretical model to the following type of animal conflicts: a member of a group is at risk and needs the help of another member to be saved. As long as assistance is not provided, this individual has a positive, time-dependent rate of dying. Assisting the individual which is at risk accrues a cost, but losing it decreases each member's inclusive fitness. A potential helper's interval between the moment a group member gets into trouble and the moment it assists is a random variable, hence its strategy is to choose the distribution of this random variable. In the asymmetric conflict all the potential helpers have identical strategy sets, but each plays a different role. For example, male or female and young or old. We consider both payoff-irrelevant asymmetry and payoff-relevant asymmetry and characterize each role's stable replies. The evolutionarily stable strategies (ESS) are computed, and the model is applied to the n brothers' problem. According to our results immediate assistance and no assistance are possible ESS both under payoff-relevant asymmetry and under payoff-relevant asymmetry. PMID- 15530497 TI - Connection between stochastic and deterministic modelling of microbial growth. AB - We present in this paper various links between individual and population cell growth. Deterministic models of the lag and subsequent growth of a bacterial population and their connection with stochastic models for the lag and subsequent generation times of individual cells are analysed. We derived the individual lag time distribution inherent in population growth models, which shows that the Baranyi model allows a wide range of shapes for individual lag time distribution. We demonstrate that individual cell lag time distributions cannot be retrieved from population growth data. We also present the results of our investigation on the effect of the mean and variance of the individual lag time and the initial cell number on the mean and variance of the population lag time. These relationships are analysed theoretically, and their consequence for predictive microbiology research is discussed. PMID- 15530498 TI - Endobain E, a brain Na+, K+ -ATPase inhibitor, decreases norepinephrine uptake in rat hypothalamus. AB - The ability of an endogenous brain Na+, K+ -ATPase inhibitor, termed endobain E, to increase [3H]norepinephrine release in rat hypothalamus was previously reported. Endobain E effect on neurotransmitter uptake was studied by assaying [3H]norepinephrine uptake in rat hypothalamus preparations, to observe uptake inhibition, which reached 60% with endobain E equivalent to 100 mg fresh cerebral cortex, an effect achieved with 40 or 400 microM ouabain. Results support the proposal that endobain E behaves as an ouabain-like substance. Taken jointly results obtained on neurotransmitter release and uptake, the suggestion that endobain E may enhance norepinephrine availability in the synaptic gap and thus lead to an increase in noradrenergic activity is advanced. PMID- 15530499 TI - Small conductance calcium-activated K+ channels, SkCa, but not voltage-gated K+ (Kv) channels, are implicated in the antinociception induced by CGS21680, a A2A adenosine receptor agonist. AB - It has been shown that A2A adenosine receptors are implicated in pain modulation. The precise mechanism by which activation of A2A receptors produces analgesic effects, however, remains unclear. The aim of this study was to investigate the possible involvement of apamin-sensitive calcium-activated potassium channels (SKCa) and voltage-gated potassium (Kv) channels in A2A receptor activation induced analgesic effects. Using mice, we evaluated the influence of apamin, a non specific blocker of SKCa channels, Lei-Dab7 (an analog of scorpion Leiurotoxin), a selective blocker of SKCa2 channels, and kaliotoxin (KTX) a Kv channel blocker, on the CGS 21680 (A2A adenosine receptor agonist)-induced increases in hot plate and tail pinch latencies. All drugs were injected in mice via the intracerebroventricular route. We found that apamin and Lei-Dab7, but not KTX, reduced antinociception produced by CGS21680 on the hot plate and tail pinch tests in a dose dependent manner. Lei-Dab 7 was more potent than apamin in this regard. We conclude that SKCa but not Kv channels are implicated in CGS 21680 induced antinociception. PMID- 15530500 TI - Specificity and sensitivity of S100B levels in amniotic fluid for Down syndrome diagnosis. AB - Down syndrome (DS) is the most common chromosomal abnormality and is associated with an extra copy of the chromosome 21. Although several markers are commonly used during pregnancy for the screening of DS, the definitive diagnosis is based on karyotype after amniocentesis, which is an expensive and laborious analysis. S100B is an astrocyte protein which had its gene mapped to the long arm of chromosome 21. Previous preliminary reports have found increased levels of this protein in the amniotic fluid of DS gestations. Aiming to achieve a simpler and cheaper test then karyotype to perform prenatal diagnosis of DS, here we have extended our previous studies and evaluated the real usefulness of amniotic S100B measurement for prenatal DS diagnosis. We have measured S100B in amniotic fluid of 96 pregnancies with DS and of 50 normal pregnancies. Pregnancies with DS presented significantly higher amniotic fluid S100B levels (M = 1.16 ng/mL; IQ = 0.83/1.78) than normal pregnancies (M = 0.51 ng/mL; IQ = 0.38/0.83) (p < 0.0001). A receiver operating characteristic (ROC) curve was performed to evaluate the sensitivity and specificity of S100B for DS diagnosis, and presented an area under the curve (AUC) of 0.82, indicating that S100B could be a reliable marker of DS. Moreover, values above 1.67 ng/mL were present only in DS fetuses, representing about 30% of affected pregnancies. However, as an overlap of values was observed between normal and DS gestations, we concluded that amniotic S100B alone is not a good test to discard DS diagnosis. PMID- 15530501 TI - Pharmacological basis of 'Yin-nourishing' and 'Yang-invigorating' actions of Cordyceps, a Chinese tonifying herb. AB - Cordyceps sinensis (Berk.) Sacc. (Cordyceps), a popular Chinese tonifying herb, was revered for being both 'Yin-nourishing' and 'Yang-invigorating' in Chinese medicine. In order to establish the pharmacological basis for the 'Yin nourishing' and 'Yang-invigorating' action of Cordyceps, the effects of wild and cultured Cordyceps on concanavalin A (Con A)-stimulated splenocytes, an in vitro bioassay for 'Yin-nourishment', and myocardial ATP generation capacity, an ex vivo bioassay for 'Yang-invigoration', were investigated in mice. The results indicated that methanolic extracts of wild and cultured Cordyceps enhanced both the Con A-stimulated splenocyte proliferation in vitro and myocardial mitochondrial ATP generation ex vivo in mice, with no significant difference in potency of action between the two types of Cordyceps. While the immuno potentiating effect was associated with the increase in interleukin II production, the stimulation of myocardial ATP generation was paralleled by an enhancement in mitochondrial electron transport. When compared with typical 'Yin' and 'Yang' tonifying Chinese herbs, Cordyceps was found to possess both 'Yin nourishing' and 'Yang-invigorating' activities, with a lower potency in both modes of action. The pharmacological characterization of Cordyceps by means of contemporary bioassays is consistent with the time-honored clinical observation from Chinese herbalists. PMID- 15530502 TI - Obstructive cholestasis alters intestinal transit in mice: role of opioid system. AB - Acute cholestasis is associated with increased activity of the endogenous opioid system. It is also known that opioid receptor agonists like morphine decrease the intestinal transit. The purpose of the present study was to investigate the effect of cholestasis on the small intestine transit and the possible involvement of opioid system in this phenomenon in mice. Cholestasis was induced by bile duct ligation and intestinal transit was measured with charcoal meal and calculation of percent of transit through small intestine. The effect of chronic administration of naltrexone and acute pretreatment with morphine on intestinal transit was evaluated in bile duct-ligated (BDL) as well as unoperated (CTL) and sham-operated (SHAM) animals. The plasma alkaline phosphatase and alanine aminotransferase activities were also measured. A significant decrease in small intestine transit (%transit) was observed in BDL mice compared to SHAM animals, which was prominent even after 24 h of cholestasis. Chronic pretreatment with an opioid receptor antagonist, naltrexone, (10 mg/kg, i.p for 2, 4 or 6 days) completely restored the cholestasis-induced decrease in %transit to that of control animals. Although the acute administration of morphine (2 mg/kg, s.c.) 20 min before charcoal feeding caused a significant decrease in the intestinal transit of CTL and SHAM animals, it did not decrease the %transit of BDL animals on the day 5 after operation. Our findings show that acute cholestasis is associated with a prominent decrease in small intestine transit in mice and opioid receptors maybe involved in this phenomenon. PMID- 15530503 TI - Effects of interferon-alpha on cloned opioid receptors expressed in Xenopus oocytes. AB - Interferon-alpha (IFNalpha) affects the opioid system. However, the direct action of IFNalpha on cloned opioid receptors remains unknown. Taking advantage of the functional coupling of cloned opioid receptors to G protein-activated inwardly rectifying K+ (GIRK) channels in a Xenopus oocyte expression system, we investigated the effects of recombinant IFNalpha on cloned mu-, delta- and kappa opioid receptors. In oocytes co-injected with mRNAs for either the delta- or kappa-opioid receptor and for GIRK channel subunits, IFNalpha at high concentrations induced small GIRK currents that were abolished by naloxone, an opioid-receptor antagonist, compared with the control responses to each selective opioid agonist. Additionally, IFNalpha induced no significant current response in oocytes injected with mRNA(s) for either opioid receptor alone or GIRK channels. In oocytes expressing the mu-opioid receptor and GIRK channels, IFNalpha had little or no effect. Moreover, in oocytes expressing each opioid receptor and GIRK channels, GIRK current responses to each selective opioid agonist were not affected by the presence of IFNalpha, indicating no significant antagonism of IFNalpha toward the opioid receptors. Furthermore, IFNalpha had little or no effect on the mu/delta-, delta/kappa- or mu/kappa-opioid receptors expressed together with GIRK channels in oocytes. Our results suggest that IFNalpha weakly activates the delta and kappa-opioid receptors. The direct activation of the delta- and kappa-opioid receptors by IFNalpha may partly contribute to some of the IFNalpha effects under its high-dose medication. PMID- 15530504 TI - Alpha1 adrenoceptor subtypes in human urinary bladder: sex and regional comparison. AB - A detailed study of the presence of alpha1 AR binding sites and alpha1 AR subtype mRNA expression in human urinary bladder areas involved in the micturition (i.e. detrusor, trigone and neck) is reported here, investigating whether or not there are differences between sexes. Results obtained indicated that alpha1 AR proteins were detectable in each bladder area. In both sexes, the detrusor and the neck expressed similar levels of alpha1 ARs: respectively, detrusor: 14.6 +/- 1.2 in men and 13.1 +/- 1.1 fmol/mg prot in women; neck: 16.9 +/- 3.2 in men and 17.5 +/ 4.1 fmol/mg prot in women. In the trigone, significantly higher alpha1ARs were found in women compared to men (20.6 +/- 1.1 vs 11.7 +/- 0.7 fmol/mg prot). Subtype analysis indicated that in women, each area was endowed with mRNA encoding for each alpha1 AR subtype. The men detrusor expressed alpha1a and alpha1d ARs, while in the trigone and the neck, each subtype was present. Since the detrusor muscle hypertrophy is a marker of bladder obstructive outlet, the selective alpha1 AR subtype targeting arouses much interest, as evidence indicates that there are differences in signalling pathways among the subtypes. Furthermore, the significance of the alpha1 ARs coexpression is still unknown; interestingly, recent papers demonstrate that alpha1 AR subtypes could dimerize. Thus, in the human urinary bladder it may be suggested a potential level of alpha1 AR complexity that could have an impact on drug development. PMID- 15530505 TI - 5-HT activates ERK MAP kinase in cultured-human peripheral blood mononuclear cells via 5-HT1A receptors. AB - In the present work, we tested the hypothesis that serotonin (5-hydroxytryptamine = 5-HT) might activate the extracellular signal-regulated kinase (ERK) pathway in human peripheral blood mononuclear cells (PBMC). PBMC were maintained in culture for 72 hrs at 37 degrees C prior to the addition of 5-HT. Our results showed an increase in ERK activation by 5-HT with a peak effect at 30 min and maximal stimulation with 5-HT at 1microM. This activation of ERK did not occur in adherent monocytes suggesting that the effect was on lymphocytes. In addition, p38 MAP kinase was not activated under these conditions. The effect of 5-HT on ERK activation appeared to be mediated through the activation of 5-HT1A receptors since similar results were obtained with R-+-8-hydroxy-DPAT, a selective 5-HT1A receptor agonist and WAY100635, a selective 5-HT1A receptor antagonist, reversed the 5-HT and the R-+-8-hydroxy-DPAT effects. Results from Western blot analysis confirmed the presence of 5-HT1A receptors on the PBMC. A 5-HT2A antagonist, ketanserin, and a 5-HT transport inhibitor, fluoxetine, both failed to block the activation of ERK by 5-HT. Our results indicate that 5-HT activates ERK, but not p38, MAP kinase of human PBMC via a 5-HT1A receptor. PMID- 15530506 TI - Interaction between the polyol pathway and non-enzymatic glycation on aortic smooth muscle cell migration and monocyte adhesion. AB - We investigated for the interaction between the polyol pathway and enhanced non enzymatic glycation, both implicated in the pathogenesis of diabetic atherosclerosis, in the activation of aortic smooth muscle cell (SMC) function. Mouse aortas and primary cultures of SMCs from wildtype (WT) mice and transgenic (TG) mice expressing human aldose reductase (AR) were studied regarding changes in AR activity, and SMC gene activation, migration and monocyte adhesion, in response to advanced glycation end-product modified BSA (AGE-BSA). Results showed that AGE-BSA increased AR activity in both WT and TG aortas, with greater increments (p < 0.05) in TG aortas which, basally, had elevated AR activity (2.8 fold of WT). These increments were attenuated by zopolrestat, an AR inhibitor. Similar AGE-induced increments in AR activity were observed in primary cultures of aortic SMCs from WT and TG mice (60% and 100%, respectively, P < 0.01). Such increments were accompanied by increases in intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) mRNA levels (both P < 0.05), activation of membrane-associated PKC-beta1 (P < 0.05) as well as increased SMC migration and Tamm-Horsfall protein (THP)-1 monocyte adhesion to SMCs (both p < 0.01), with all changes being significantly greater in TG SMCs (P < 0.05) and suppressible by either zopolrestat or transfection with an AR antisense oligonucleotide. Our findings suggest that the effects of AGEs on SMC activation, migration and monocyte adhesion are mediated partly through the polyol pathway and, possibly, PKC activation. The greater AGE-induced changes in the TG SMCs have provided further support for the dependency of such changes on polyol pathway hyperactivity. PMID- 15530507 TI - Lipopolysaccharide-induced pulmonary inflammation is not accompanied by a release of anandamide into the lavage fluid or a down-regulation of the activity of fatty acid amide hydrolase. AB - The effect of lipopolysaccharide inhalation upon lung anandamide levels, anandamide synthetic enzymes and fatty acid amide hydrolase has been investigated. Lipopolysaccharide exposure produced a dramatic extravasation of neutrophils and release of tumour necrosis factor alpha into the bronchoalveolar lavage (BAL) fluid, which was not accompanied by epithelial cell injury. The treatment, however, did not change significantly the levels of anandamide and the related compound palmitoylethanolamide in the cell-free fraction of the BAL fluid. The activities of the anandamide synthetic enzymes N-acyltransferase and N acylphosphatidylethanolamine phospholipase D and the activity of fatty acid amide hydrolase in lung membrane fractions did not change significantly following the exposure to lipopolysaccharide. The non-selective fatty acid amide hydrolase inhibitor phenylmethylsulfonyl fluoride was a less potent inhibitor of lung fatty acid amide hydrolase than expected from the literature, and a dose of 30 mg/kg i.p. of this compound, which produced a complete inhibition of brain anandamide metabolism, only partially inhibited the lung metabolic activity. PMID- 15530508 TI - Feeding response to ghrelin agonist and antagonist in lean and obese Zucker rats. AB - Ghrelin is a new orexigenic and adipogenic peptide primarily produced by the stomach and the hypothalamus. In the present experiment, we determined the circulating ghrelin levels in 60-week old fa/fa Zucker rats with a well established obesity (n = 12) and in their lean (FA/FA) counterparts (n = 12). We also tested the feeding response of both groups to intra-peritoneal (I.P.) injection of ghrelin agonist and antagonist. Obese rats ate significantly more than the lean rats (21.7 +/- 1.1 vs. 18.3 +/- 0.3 g/day; p < 0.01). Their plasma ghrelin concentration was 35% higher than that in the lean homozygous rats (p < 0.025). GHRP-6 (1 mg/kg I.P, a GHS-R agonist) stimulated food intake in lean but not in obese rats (p < 0.01), whereas [D-Lys)]-GHRP-6 (12 mg/kg I.P., a GHS-R antagonist) decreased food intake in both groups (p < 0.0001). These results indicate that the obese Zucker rat is characterized by an increase in plasma ghrelin concentrations and by an attenuated response to a GHS-R agonist. They support a role for ghrelin in the development of obesity in the absence of leptin signaling. PMID- 15530509 TI - Marine habitats: loss and gain, mitigation and compensation. PMID- 15530510 TI - Science, policy and the management of sewage materials. The New York City experience. AB - Development of national policy on sewage sludge management is a classic example of incremental policy formulation [Fiorino, D.J. 1995. Making Environmental Policy. University of California Press. Berkeley, CA. p. 269]. Consequently, policy has developed piecemeal, and results are, in some ways, different than intended. Land application of sewage sludge has not been a panacea. Many of the same types of policy are now being raised about it. We demonstrate this by examining the management of sewage materials by New York City from near the turn of the 20th century, when ocean dumping was viewed as a means to alleviate some of the gross pollution in New York Harbor, to when ocean dumping was banned, and thence to the present when sludge is applied to land as "biosolids." Lessons learned during this long, sometimes contentious history can be applied to present situations--specifically not understanding the long-term consequences of land based reuse and disposal technologies. PMID- 15530511 TI - A uniform terminology on bioinvasions: a chimera or an operative tool? AB - A consensual set of definitions regarding bioinvasions is essential in order to facilitate discourse among the science, policy and management communities dealing with the issue. Considering both the mode of entry and the extent of the impact of an alien species into a new environment, a set of key terms is proposed as an operative tool for marine scientists. PMID- 15530512 TI - Genetic variation among age-classes of the mangrove Avicennia marina in clean and contaminated sediments. AB - An examination was made of the genetic variation with age in mangrove populations in two estuaries in south-east Australia, one contaminated, the other uncontaminated. Sediment nutrient and metal levels, and isozyme/allozyme complements were measured in three age-classes in four stands in each estuary, to identify site and intergenerational genetic composition, and influences of sediment contamination. Within each river, there were greater genetic distances among the age-classes than among sampling sites. A downstream gradient of genetic distance was found in the uncontaminated river, but not the contaminated one, where genetic distances seemed to reflect site-specific factors, though apparently not the present sediment metal or nutrient levels. It appears that genetic distances in the polluted river may reflect past fluctuations in pollution pressures. In both rivers, genetic distances indicate that the mangroves within one estuary form one population, with 'neighbourhood' differentiation. PMID- 15530513 TI - Monitoring chemical contamination levels in the Mediterranean based on the use of mussel caging. AB - Within the framework of the biointegrator network (RINBIO), 92 man-made cages containing mussels (Mytilus galloprovincialis), distributed over 1800 km of the French Mediterranean coast, made it possible to assess chemical contamination by heavy metals (Cd, Hg, Zn, Pb, Cu, Ni, Cr, As) and organic compounds (DDT, PCBs, HAP). The caging technique was adopted to compensate for the scarcity of natural shellfish stocks in the Mediterranean and to enable comparison of the sites regardless of their physicochemical and trophic characteristics. Models linking a biometric parameter (the condition index) to pollutant levels make it possible to adjust raw data on contamination for a reference individual, by making a clear distinction between physiological factors (growth) and environmental ones. The results demonstrate significant levels of pollutants in sites heretofore unsampled; the pollution occurs at limited areas, and the sources have been identified. Moreover, average contamination levels are comparable to those measured in wild species and in other areas. Overall, the research demonstrates the reliability of this methodological approach for monitoring marine pollution, especially in the Mediterranean. PMID- 15530514 TI - Seasonal variations of physiological and cellular biomarkers and their use in the biomonitoring of north Adriatic coastal waters (Croatia). AB - Lysosomal membrane stability, the content of neutral lipids and lipofuscin as cellular biomarkers were measured in the digestive gland of mussels, Mytilus galloprovincialis Lam., during a one year survey at selected sites along the coast of the northern Adriatic. The ability of mussels to survive in air as a general physiological response was tested at the same sites. Annual changes of sea temperature, salinity, O(2) saturation, pH and chlorophyll a concentration were measured and the effects of these environmental factors on the biomarkers selected for this study were determined. Survival of mussels after aerial exposure was significantly (p < 0.05) reduced in mussels collected from the contaminated harbors of Pula and Rijeka compared to clean control sites. Lysosomal membrane stability of mussels from urban and industrial areas was also lower indicating exposure to different environmental contaminants. At the same sites, the content of neutral lipids and lipofuscin in digestive cells was significantly higher compared to the reference sites, suggesting alterations in lipid metabolism and contaminant-induced peroxidation of cell membranes. The greatest influence of natural environmental factors was observed for the aerial survival of mussels and the neutral lipids and lipofuscin content in the digestive gland, indicating that the annual cycles of biomarker responses should be well understood for a correct interpretation of biomonitoring results. PMID- 15530515 TI - Effects of raised CO2 concentration on the egg production rate and early development of two marine copepods (Acartia steueri and Acartia erythraea). AB - Direct injection of CO(2) into the deep ocean is receiving increasing attention as a way to mitigate increasing atmospheric CO(2) concentration. To assess the potential impact of the environmental change associated with CO(2) sequestration in the ocean, we studied the lethal and sub-lethal effects of raised CO(2) concentration in seawater on adult and early stage embryos of marine planktonic copepods. We found that the reproduction rate and larval development of copepods are very sensitive to increased CO(2) concentration. The hatching rate tended to decrease, and nauplius mortality rate to increase, with increased CO(2) concentration. These results suggest that the marine copepod community will be negatively affected by the disposal of CO(2). This could decrease on the carbon export flux to the deep ocean and change the biological pump. Clearly, further studies are needed to determine whether ocean CO(2) injection is an acceptable strategy to reduce anthropogenic CO(2). PMID- 15530516 TI - Marine quality assessment by use of benthic species-abundance distributions: a proposed new protocol within the European Union Water Framework Directive. AB - The aim of this study is to develop a new method for classification of marine benthic quality according to the European Union Water Framework Directive. Tolerance values to environmental disturbance were determined in an objective analysis for benthic species along the Swedish west coast by using 4676 samples from 257 stations. Based on a combination of the species tolerance values, abundance and diversity, a benthic quality index (BQI) was calculated for the assessment of environmental status at a particular station. The qualification of BQI was evaluated in relation to known spatial and temporal gradients of disturbance. PMID- 15530517 TI - Response of the bacterial community to in situ bioremediation of organic-rich sediments. AB - A field trial experiment was carried out to assess the potential of bioremediation for mobilisation of carbon in organic-rich sediments. Both bioaugmentation (bio-fixed microorganisms) and biostimulation (oxygen release compounds--ORC) protocols have been tested and the response of the bacterial community has been described to assess the baseline for bioremediation potential. Multifactorial ANOVA revealed that bioaugmentation protocol had an effect in stimulate mobilisation processes and significantly enhanced extra-cellular enzymatic activity rates. In contrast biostimulation treatment did not have an effect on mobilisation rates but contributed to enhance bacterial efficiency through a maximization of the bacterial production:enzymatic activity ratio. Average calculation of net mobilised carbon showed that 23% increase of mobilised pool was accounted for bioaugmentation in summer. Although biostimulation accounted for a smaller increase in mobilised carbon (<10%), the use of ORC resulted in an increased mineralisation and net carbon loss via respiration. Based on our results, a conceptual model for application of bioremediation to face the problem of sediment eutrophication is discussed. PMID- 15530518 TI - Microbiological Indices for total coliform and E. coli bacteria in estuarine waters. AB - Bacterial counts for total coliforms and E. coli in estuaries are normally orders of magnitude greater at the freshwater end than at the seaward end and tidal movements and variations in freshwater flows produce continual change in the freshwater/seawater mix--this causes the bacterial counts to vary greatly throughout the estuary and the complexity creates difficulty in appraising or assessing the bacterial counts (i.e. difficulties arise when deciding if the bacterial counts for estuarine water samples are within an acceptable range- relative to their corresponding salinities). The situation is further complicated in estuaries where sewage is discharged directly. Microbiological criteria and indices that can be used in a practical way to overcome these difficulties were developed. The procedure is summarised as follows: 1. Primary criteria are proposed for total coliform and for E. coli bacteria in the freshwater at the head of the estuary and in full seawater at the mouth of the estuary. 2. For estuarine or transitional waters (i.e. waters with salinity ranging from 0 per thousand to 35 per thousand), distinct criteria are calculated for each individual sample--with a separate criterion for total coliforms and for E. coli- the individual criteria are founded on the primary criteria and vary with salinity on a pro-rata or linear basis. 3. Finally, the Microbiological Index for each result is obtained by dividing the actual bacterial count by the corresponding criterion--the acceptable Index is then equal to 1--at any salinity (i.e. where the Index is <1 then the bacterial count complies with the criterion, and where the Index is >1 then the count breaches the criterion). The Index also indicates the extent of the compliance or non-compliance with the corresponding criteria. An example of the application of the Microbiological Index is also presented--including examples of graphs that demonstrate how microbiological data for estuarine waters can be summarised and presented. PMID- 15530519 TI - Investigation of natural and anthropogenic hydrocarbon inputs in sediments using geochemical markers. I. Santos, SP--Brazil. AB - A study on the inputs of organic matter to marine sediments of Santos Bay and Estuary, located in southeastern Sao Paulo State, Brazil, is presented here. The following geochemical markers were identified and quantified: aliphatic hydrocarbons (normal and isoprenoid alkanes), petroleum biomarkers, linear alkylbenzenes (LABs) and polycyclic aromatic hydrocarbons (PAHs). Sediment samples, collected at 15 sites in the bay and surrounding channels, were extracted and analyzed by gas chromatography with flame ionization and mass spectrometer detectors. Total concentrations varied from 0.17 to 107.80 microg g( 1) for aliphatics, from 47.2 to 2560.1 ng g(-1) for petroleum biomarkers, from 16.9 to 430.6 ng g(-1) for LABs and from 79.6 to 15389.1 ng g(-1) for PAHs. Anthropogenic contributions were apparent, indicating the influences of the Cubatao industrial complex, sewage outfalls along the area, the waste deposit of Santos city and storage facilities for petroleum and derivatives, based on the higher concentrations of geochemical markers detected. PMID- 15530520 TI - Shore litter along sandy beaches of the Gulf of Oman. AB - Beach debris abundance and weight were estimated from surveys on 11 beaches of the Gulf of Oman along the Omani coast. Debris were collected on two occasions from 100 m transects, sorted and categorized by origin and type. Overall contaminations ranged from 0.43 to 6.01 items m(-1) of beach front on different beaches with a mean value of 1.79+/-1.04 gm(-1) (95% C.I). In terms of weight, contamination levels ranged from 7.8 to 75.44 gm(-1) of beach front with a mean contamination of 27.02+/-14.48 gm(-1) (95% C.I). In terms of numbers of items, plastic debris ranked first on all beaches followed by either wood items or other organic materials such as cigarette butts. Industrial debris remained few on all beaches (<10%). Most debris had a local origin and, in terms of numbers, were associated with beach recreational activities whereas fishing debris represented the largest proportion of the debris in terms of weight. There were notable differences between beaches in the relative abundance of recreation-related and fishing-related debris. PMID- 15530521 TI - Role of container vessels in the introduction of exotic species. AB - Ballast water exchange practices were monitored on 28 incoming container vessels at the Port of Montreal. Measurements on 15 vessels indicated 13 of 32 tanks had salinities of <30 per thousand. The 16 transits with a North Atlantic route visited 31 of 37 ports located on freshwater or near freshwater outflows. Ballast carried by this vessel type represents an important means for the introduction of species on a global scale because of its transit routes, dockside discharge and moving ballast between tanks. Container vessels represent about 15% of the world fleet, but account for 32% of all visits to global ports, and 46% of visits to the 25 largest ports. The 10 ports that handled the largest volumes of international cargo also included 8 that handled the most cargo containers. Large ports can receive over 100,000 visits by all vessel types annually, and serve as hubs for over 500 ports in 100 countries. Secondary transport of exotic species is also a concern because of frequent visits by regional vessels. PMID- 15530522 TI - 2,4,6-trinitrotoluene transformation by a tropical marine yeast, Yarrowia lipolytica NCIM 3589. AB - Yarrowia lipolytica NCIM 3589, a tropical marine degrader of hydrocarbons and triglycerides transformed 2,4,6-trinitrotoluene (TNT) very efficiently. Though this yeast could not utilize TNT as the sole carbon or nitrogen source, it was capable of reducing the nitro groups in TNT to aminodinitrotoluene (ADNT). In a complete medium containing glucose and ammonium sulphate as the available carbon and nitrogen sources respectively, the culture was able to completely transform 1 mM (227 ppm) of TNT under such conditions. A dual pathway was found to be functional, one of which resulted in the formation of the hydride-Meisenheimer complex (H(-)TNT) as a transiently accumulating metabolite that was subsequently denitrated to 2,4-dinitrotoluene (2,4-DNT), whereas the other pathway resulted in the formation of amino derivatives. The presence of increasing amounts of reducing equivalents in the form of glucose promoted better growth and the nitroreductases of this yeast to reduce the aromatic ring to 2,4-DNT although, the reduction of the nitro groups to amino groups was the major functional pathway. The ability of this tropical marine yeast to transform TNT into products such as 2,4-DNT which in turn could be metabolized by other microbes has implications in the use of this yeast for bioremediation of TNT polluted marine environments. PMID- 15530523 TI - Concentrations of trace elements in muscle of sturgeons in the Caspian Sea. AB - Concentrations of 21 trace elements were determined in muscle of beluga (Huso huso), Persian sturgeon (Acipenser persicus), Russian sturgeon (Acipenser gueldenstaedtii), ship sturgeon (Acipenser nudiventris) and stellate sturgeon (Acipenser stellatus) collected from coastal regions of Azerbaijan, Iran, Kazakhstan and Turkmenistan, surrounding the Caspian Sea during 2000-2001. Concentrations of Mn, Co, Cu, Mo, Sn, Hg, Pb and Bi in the muscle were apparently different among the five species of sturgeons. Especially, beluga showed the highest concentrations of Hg, Pb and Mn in all the five species. In addition, more than half of the individuals of beluga exceeded the guideline level (0.3 microg/g wet wt.) of Hg for food in UK. However, V, Cr, Zn, Ga, Rb, Sr and Ba concentrations were similar among five sturgeons. Growth-dependent increase in Hg in beluga, Cu in Persian sturgeon, and Zn in Russian sturgeon were observed, whereas V, Mn, Co, Cu, Ga, Sr, Mo, Sn, Hg and Bi concentrations in Persian sturgeon, Pb in ship sturgeon, and Cr, Mn, and Rb in stellate sturgeon were negatively correlated with body length. All species of sturgeon in Azerbaijan showed the highest concentration of Sr and the lowest concentration of Rb, while the five sturgeons from Iran showed opposite trends. Concentration of V, which is present in oil, in sturgeons in the Caspian Sea was considerably lower than that of fish from Kuwait, but was comparable to that of Cambodia and the Gulf and Gulf of Oman. To our knowledge, this study provides the first extensive data on multielemental accumulation in sturgeons of the Caspian Sea. PMID- 15530524 TI - Total oxyradical scavenging capacity responses in Mytilus galloprovincialis transplanted into the Venice lagoon (Italy) to measure the biological impact of anthropogenic activities. AB - Oxidative stress related investigations to monitor the impact of the pollutant discharges into the Venice lagoon (Italy) originating from anthropogenic activities (raw sewage water, agricultural and industrial effluents, oil tanker traffic), on marine organisms have classically been carried out by analyzing specific, single antioxidants (i.e. catalase, superoxide dismutase). In this paper, two studies are reported where the total oxyradical scavenging capacity assay (TOSC) was selected and measured toward peroxyl, and hydroxyl free radicals, and peroxynitrite in mussels (Mytilus galloprovincialis) transplanted into the Venice city and throughout the lagoon to measure the biological effects of anthropogenic activities. In the first experiment, mussels from a clean site (farm) were transplanted to the urban area of Venice for 0, 1, 2 and 4 weeks; cytosolic TOSC toward peroxyl and hydroxyl free radicals, and peroxynitrite revealed that the transplantation process caused a stress (handling stress, anoxia, oxidative burst) resulting in a reduction of TOSC in both control and urban sites, therefore, preventing clear interpretation of the data after one week. At week 2, a significant TOSC reduction (P < 0.05) toward peroxyl and hydroxyl radicals in the urban site revealed that mussels experienced oxidative pressure exerted by pollutants. Most TOSC values returned to initial levels at week 4; however, TOSC induction was noticed in the control group toward peroxyl and hydroxyl radicals while in the exposed group it was not indicating an inhibition of the oxidative metabolism. In the second experiment, mussels were deployed at seven different sites throughout the lagoon. After five weeks of exposure, significant TOSC reduction was measured (P < 0.05) toward peroxyl for Palude della Rosa, Chioggia and Valle Millecampi, toward hydroxyl radicals for Valle Millecampi and Campalto and toward peroxynitrite for Valle Millecampi. Although these data indicate a depletion of the low molecular weight scavengers, additional biomarkers are needed to draw a conclusion on the health of the mussels. TOSC was proved to be an interesting health index parameter to measure pollution impact in a transplantation study provided that the mussels are exposed for two weeks and a control is run in parallel. PMID- 15530525 TI - Source apportionment of trace metals in surface waters of a polluted stream using multivariate statistical analyses. AB - Surface water samples were collected from ten previously selected sites of the polluted Dil Deresi stream, during two field surveys, December 2001 and April 2002. All samples were analyzed using ICP-AES, and the concentrations of trace metals (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Pb, Sn and Zn) were determined. The results were compared with national and international water quality guidelines, as well as literature values reported for similar rivers. Factor analysis (FA) and a factor analysis-multiple regression (FA-MR) model were used for source apportionment and estimation of contributions from identified sources to the concentration of each parameter. By a varimax rotated factor analysis, four source types were identified as the paint industry; sewage, crustal and road traffic runoff for trace metals, explaining about 83% of the total variance. FA MR results showed that predicted concentrations were calculated with uncertainties lower than 15%. PMID- 15530526 TI - The community composition and production of phytoplankton in fish pens of Cape Bolinao, Pangasinan: a field study. AB - From 1995 up to the present, fish pens proliferated in the municipal waters of Bolinao, northern Philippines. Since then, fish kills and phytoplankton blooms have been recurrent. Have fishpens altered the phytoplankton community composition and production of these waters? The phytoplankton community in Cape Bolinao, Lingayen Gulf is typical of a tropical coastal area where diatoms alternate with dinoflagellates during the dry and wet seasons. In the nutrient rich fish pens, phytoplankton in this study showed a lower diatom/dinoflagellate ratio and unusually high phytoplankton counts of 10(4) cells/l and even as high as 10(5) cells/l. Correlations between physico-chemical parameters, phytoplankton production and community composition were made in 2001. This paper tried to explain the occurrence of a Cylindrotheca closterium bloom (10(5) cells/l), during the dry season of the same year and a Prorocentrum minimum bloom (4.7 x 10(5) cells/l), which accompanied a massive fish kill during January 2002. PMID- 15530527 TI - Investigation of recreational boats as a source of copper at anchorage sites using time-integrated diffusive gradients in thin film and sediment measurements. AB - Antifouling paints on small to medium recreational vessels were first recognized as an important source of pollution in the 1970s. One of the principle biocides in these paints is Cu. Results from a field program in the Gold Coast Broadwater, Queensland, demonstrate a clear correlation between recreational boat numbers at anchorage sites and water column Cu concentrations, for both time-integrated 24 h DGT measurements (n=14, r=0.815, p<0.001) and measurements on composite 0.45 mum filtered grab samples (n=14, r=0.698, p<0.01) collected every 4 h over 24 h. At boat numbers above 30 the 0.45 microm-filterable Cu concentrations were mostly above the relevant guideline value (1.3 microgl(-1)) but the DGT-reactive Cu concentrations were well below this value at all boat numbers studied. For three day DGT deployments in Moreton Bay, Queensland, correlations between Cu levels and vessel numbers were not observed, possibly because of uncertain estimates of boat numbers. However, using a multi-factorial ANOVA, DGT-reactive Cu concentrations showed a significant effect for 'sites with vessels vs. sites without vessels' (alpha=0.10, p=0.077) and for 'sampling period' (non-holiday weekdays, weekends, holiday weekdays, holiday weekends) at alpha=0.10, p=0.02. Cu levels in sediments at the same sites were strongly influenced by fraction of clay (<63 microm) material but also by whether the samples were collected at an anchorage or control site. Results from this study further support the view that Cu emissions from antifouling paints may become an important source in waters with high boat numbers and should be taken into account when designing management instruments for coastal waterways. PMID- 15530528 TI - Eradication success down under: heat treatment of a sunken trawler to kill the invasive seaweed Undaria pinnatifida. AB - Eradication of invasive species is difficult in the marine environment, and there have only been a few successes. We report the successful eradication of the invasive seaweed Undaria pinnatifida from a sunken trawler in the Chatham Islands, New Zealand. New heat-treatment methods were developed as the most cost effective and environmentally acceptable option to kill Undaria. Monitoring of the trawler for three years after it sank found no Undaria after the vessel was treated. Key factors in the success of the eradication programme included: early detection, a rapid response, pre-existing knowledge of Undaria, an adaptive management approach, targeting of multiple life history stages, and the cooperation of the vessel's insurer. PMID- 15530529 TI - Reducing sewage pollution in the Antarctic marine environment using a sewage treatment plant. AB - Despite Antarctica being the largest pristine wilderness on Earth, many coastal Antarctic research stations release untreated sewage waste into the marine environment, which may have negative effects on local wildlife. In February 2003 a sewage treatment plant was installed at Rothera Research Station (Adelaide Island, Antarctic Peninsula). After one year of operation the sewage treatment plant had dramatically reduced the microbiological pollution in the near-shore marine environment around the outfall and seawater quality conformed to European Union Bathing Water Standards. PMID- 15530530 TI - Tissue distribution of metals in loggerhead turtles (Caretta caretta) stranded in the Canary Islands, Spain. PMID- 15530531 TI - Speciation of heavy metals in coastal sediments of Semarang, Indonesia. PMID- 15530532 TI - The invasive pest species Ciona intestinalis (Linnaeus, 1767) reported in a harbour in southern Western Australia. PMID- 15530533 TI - Total and organic mercury concentrations in muscle tissue of the blue shark (Prionace glauca L.1758) from the Northeast Atlantic. PMID- 15530534 TI - Abandon hope all ye target theory modelers: on the effects of low dose exposures to ionizing radiation and other carcinogens. PMID- 15530535 TI - Damaging and protective cell signalling in the untargeted effects of ionizing radiation. AB - The major adverse consequences of radiation exposures are attributed to DNA damage in irradiated cells that has not been correctly restored by metabolic repair processes. However, the dogma that genetic alterations are restricted to directly irradiated cells has been challenged by observations in which effects of ionizing radiation arise in non-irradiated cells. These, so called, untargeted effects are demonstrated in cells that are the descendants of irradiated cells either directly or via media transfer (radiation-induced genomic instability) or in cells that have communicated with irradiated cells (radiation-induced bystander effects). Radiation-induced genomic instability is characterized by a number of delayed responses including chromosomal abnormalities, gene mutations and cell death. Bystander effects include increases or decreases in damage inducible and stress-related proteins, increases or decreases in reactive oxygen and nitrogen species, cell death or cell proliferation, cell differentiation, radioadaptation, induction of mutations and chromosome aberrations and chromosomal instability. The phenotypic expression of untargeted effects and the potential consequences of these effects in tissues reflect a balance between the type of bystander signals produced and the responses of cell populations to such signals, both of which may be significantly influenced by cell type and genotype. Thus, in addition to targeted effects of damage induced directly in cells by irradiation, a variety of untargeted effects may also make important short-term and long-term contributions to determining overall outcome after radiation exposures. PMID- 15530536 TI - Interrelationships amongst radiation-induced genomic instability, bystander effects, and the adaptive response. AB - Over the past two decades, our understanding of radiation biology has undergone a fundamental shift in paradigms away from deterministic "hit-effect" relationships and towards complex ongoing "cellular responses". These responses include now familiar, but still poorly understood, phenomena associated with radiation exposure such as bystander effects, genomic instability, and adaptive responses. All three have been observed at very low doses, and at time points far removed from the initial radiation exposure, and are extremely relevant for linear extrapolation to low doses; the adaptive response is particularly relevant when exposure is spread over a period of time. These are precisely the circumstances that are most relevant to understanding cancer risk associated with environmental and occupational radiation exposures. This review will provide a synthesis of the known, and proposed, interrelationships amongst low-dose cellular responses to radiation. It also will examine the potential importance of non-targeted cellular responses to ionizing radiation in setting acceptable exposure limits especially to low-LET radiations. PMID- 15530537 TI - Phenomena leading to cell survival values which deviate from linear-quadratic models. AB - For several decades, the prevailing paradigm for modeling the effects of ionizing radiation (IR) on living systems was the target model with its inherent assumptions--that only those cells in the radiation path whose molecules sustained collisions with high energy particles and rays were damaged, that the damage was proportional to the energy absorbed by each cell and to the number of cells absorbing energy, and that all cells had identical sensitivities to radiation. However, evidence has accumulated that cells exhibit phenomena at low radiation exposures that appear to contradict at least one of these assumptions. Some of these phenomena currently under active study include low-dose hypersensitivity (HRS), increased radiation radioresistance (IRR), the adaptive response (AR), the bystander effect (BE), and death-inducing factor (DIE). These effects may interact to give rise to other phenomena such as hormesis, in which small amounts of otherwise toxic agent appear to be beneficial. Elucidating the cellular and molecular bases for these phenomena will lead to greater understanding of the relationships of these processes, including hormesis, to human health. PMID- 15530538 TI - Detection of chromosomal instability in alpha-irradiated and bystander human fibroblasts. AB - There is increasing evidence biological responses to ionizing radiation are not confined to those cells that are directly hit, but may be seen in the progeny at subsequent generations (genomic instability) and in non-irradiated neighbors of irradiated cells (bystander effects). These so called non-targeted phenomena would have significant contributions to radiation-induced carcinogenesis, especially at low doses where only a limited number of cells in a population are directed hit. Here we present data using a co-culturing protocol examining chromosomal instability in alpha-irradiated and bystander human fibroblasts BJ1 htert. At the first cell division following exposure to 0.1 and 1Gy alpha particles, irradiated populations demonstrated a dose dependent increase in chromosome-type aberrations. At this time bystander BJ1-htert populations demonstrated elevated chromatid-type aberrations when compared to controls. Irradiated and bystander populations were also analyzed for chromosomal aberrations as a function of time post-irradiation. When considered over 25 doublings, all irradiated and bystander populations had significantly higher frequencies of chromatid aberrations when compared to controls (2-3-fold over controls) and were not dependent on dose. The results presented here support the link between the radiation-induced phenomena of genomic instability and the bystander effect. PMID- 15530539 TI - Cis-acting transmission of genomic instability. AB - Genomic instability is a highly pleiotropic phenotype, which may reflect a variety of underlying mechanisms. Destabilization has been shown in some cases to involve mutational alteration or inactivation of trans-acting cellular factors, for example, p53 or mismatch repair functions. However, aspects of instability are not well explained by mutational inactivation of trans-acting factors, and other epigenetic and cis-acting mechanisms have recently been proposed. The trans and cis models result in divergent predictions for the distribution of instability-associated genetic alterations within the genome, and for the inheritance of genomic instability among sibling sub-clones of unstable parents. These predictions have been tested in this study primarily by tracking the karyotypic distribution of chromosomal rearrangements in clones and sub-clones exhibiting radiation-induced genomic instability; inheritance of mutator phenotypes was also analyzed. The results indicate that genomic instability is unevenly transmitted to sibling sub-clones, that chromosomal rearrangements within unstable clones are non-randomly distributed throughout the karyotype, and that the majority of chromosomal rearrangements associated with instability affect trisomic chromosomal segments. Observations of instability in trisomic regions suggests that in addition to promoting further alterations in chromosomal number, aneuploidy can affect the recovery of structural rearrangements. In summary, these findings cannot be fully explained by invoking a homogeneously distributed factor acting in trans, but do provide support for previous suggestions that genomic instability may in part be driven by a cis-acting mechanism. PMID- 15530540 TI - Gamma radiation-induced heritable mutations at repetitive DNA loci in out-bred mice. AB - Recent studies have shown that expanded-simple-tandem-repeat (ESTR) DNA loci are efficient genetic markers for detecting radiation-induced germline mutations in mice. Dose responses following irradiation, however, have only been characterized in a small number of inbred mouse strains, and no studies have applied ESTRs to examine potential modifiers of radiation risk, such as adaptive response. We gamma-irradiated groups of male out-bred Swiss-Webster mice with single acute doses of 0.5 and 1.0 Gy, and compared germline mutation rates at ESTR loci to a sham-irradiated control. To test for evidence of adaptive response we treated a third group with a total dose of 1.1 Gy that was fractionated into a 0.1 Gy adapting dose, followed by a challenge dose of 1.0 Gy 24h later. Paternal mutation rates were significantly elevated above the control in the 0.5 Gy (2.8 fold) and 1.0 Gy (3.0-fold) groups, but were similar to each other despite the difference in radiation dose. The doubling dose for paternal mutation induction was 0.26 Gy (95% CI = 0.14-0.51 Gy). Males adapted with a 0.1 Gy dose prior to a 1.0 Gy challenge dose had mutation rates that were not significantly elevated above the control, and were 43% reduced compared to those receiving single doses. We conclude that pre-meiotic male germ cells in out-bred Swiss-Webster mice are sensitive to ESTR mutations induced by acute doses of ionizing radiation, but mutation induction may become saturated at a lower dose than in some strains of inbred mice. Reduced mutation rates in the adapted group provide intriguing evidence for suppression of ESTR mutations in the male germline through adaptive response. Repetitive DNA markers may be useful tools for exploration of biological factors affecting the probability of heritable mutations caused by low dose ionizing radiation exposure. The biological significance of ESTR mutations in terms of radiation risk assessment, however, is still undetermined. PMID- 15530541 TI - Bystander effects, adaptive response and genomic instability induced by prenatal irradiation. AB - The developing human embryo and fetus undergo very radiosensitive stages during the prenatal development. It is likely that the induction of low dose related effects such as bystander effects, the adaptive response, and genomic instability would have profound effects on embryonic and fetal development. In this paper, I review what has been reported on the induction of these three phenomena in exposed embryos and fetuses. All three phenomena have been shown to occur in murine embryonic or fetal cells and structures, although the induction of an adaptive response (and also likely the induction of bystander effects) are limited in terms of when during development they can be induced and the dose or dose-rate used to treat animals in utero. In contrast, genomic instability can be induced throughout development, and the effects of radiation exposure on genome instability can be observed for long times after irradiation including through pre- and postnatal development and into the next generation of mice. There are clearly strain-specific differences in the induction of these phenomena and all three can lead to long-term detrimental effects. This is true for the adaptive response as well. While induction of an adaptive response can make fetuses more resistant to some gross developmental defects induced by a subsequent high dose challenge with ionizing radiation, the long-term effects of this low dose exposure are detrimental. The negative effects of all three phenomena reflect the complexity of fetal development, a process where even small changes in the timing of gene expression or suppression can have dramatic effects on the pattern of biological events and the subsequent development of the mammalian organism. PMID- 15530542 TI - Radiation-induced chromosomal instability and gene expression profiling: searching for clues to initiation and perpetuation. AB - Radiation-induced genomic instability (RIGI) manifests in the progeny of cells surviving ionizing radiation (IR), and can be measured using such endpoints as delayed mutation, micronuclei formation, and chromosomal instability. The frequency of RIGI is relatively high, exceeding the gene mutation rate of IR by orders of magnitude, leading to conjecture that a gene mutation is not the cause of the phenotype. We have started to explore whether differential gene expression patterns are associated with the instability phenotype, in order to shed light on its initiation and perpetuation. Using GM10115 human-hamster hybrid-derived chromosomally stable and radiation-induced unstable clones, gene expression patterns were analyzed using microarray analysis. Two methods were used to find differentially expressed genes, and all candidate genes identified by these methods were under-expressed relative to the chromosomally stable reference sample. Among this set differentially expressed genes identified were two candidates with a relationship to the ubiquitin/proteasome pathway. While follow up gene expression analyses have confirmed the under-expression of these two genes in some of our chromosomally unstable clones, preliminary functional studies have been unable to demonstrate a link to instability. It is anticipated that as we apply this technology to the study of radiation-induced genomic instability, clues to its onset will be revealed, ultimately contributing to a greater understanding of the mechanisms of radiation carcinogenesis. PMID- 15530543 TI - IR-inducible clusterin gene expression: a protein with potential roles in ionizing radiation-induced adaptive responses, genomic instability, and bystander effects. AB - Clusterin (CLU) plays numerous roles in mammalian cells after stress. A review of the recent literature strongly suggests potential roles for CLU proteins in low dose ionizing radiation (IR)-inducible adaptive responses, bystander effects, and delayed death and genomic instability. Its most striking and evident feature is the inducibility of the CLU promoter after low, as well as high, doses of IR. Two major forms of CLU, secreted (sCLU) and nuclear (nCLU), possess opposite functions in cellular responses to IR: sCLU is cytoprotective, whereas nCLU (a byproduct of alternative splicing) is a pro-death factor. Recent studies from our laboratory and others demonstrated that down-regulation of sCLU by specific siRNA increased cytotoxic responses to chemotherapy and IR. sCLU was induced after low non-toxic doses of IR (0.02-0.5 Gy) in human cultured cells and in mice in vivo. The low dose inducibility of this survival protein suggests a possible role for sCLU in radiation adaptive responses, characterized by increased cell radioresistance after exposure to low adapting IR doses. Although it is still unclear whether the adaptive response is beneficial or not to cells, survival of damaged cells after IR may lead to genomic instability in the descendants of surviving cells. Recent studies indicate a link between sCLU accumulation and cancer incidence, as well as aging, supporting involvement of the protein in the development of genomic instability. Secreted after IR, sCLU may also alter intracellular communication due to its ability to bind cell surface receptors, such as the TGF-beta receptors (types I and II). This interference with signaling pathways may contribute to IR-induced bystander effects. We hypothesize that activation of the TGF-beta signaling pathway, which often occurs after IR exposure, can in turn activate the CLU promoter. TGF-beta and IR-inducible de novo synthesized sCLU may then bind the TGF-beta receptors and suppress downstream growth arrest signaling. This complicated negative feedback regulation most certainly depends on the cellular microenvironment, but undoubtedly represents a potential link between IR-induced adaptive responses, genomic instability and bystander effects. Further elucidation of clusterin protein functions in IR responses are clearly warranted. PMID- 15530544 TI - Genotoxicity in the eyes of bystander cells. AB - The controversial use of a linear, no threshold extrapolation model for low dose risk assessment has become even more so in light of the recent reports on the bystander phenomenon. The answer to the question as to which of the two phenomena, bystander versus adaptive response, is more important has practical implication in terms of low dose radiation risk assessment. In this review, genotoxicity is used as an endpoint to introduce the two phenomena, provide some insight into the mechanisms of bystander effect and to bridge the two low dose phenomena which operate in opposite directions: the bystander effect tends to exaggerate the effect at low doses, by communicating damage from hit to non-hit cells whereas the adaptive response confers resistance to a subsequent challenging dose by an initial low priming dose. PMID- 15530545 TI - Radiation-induced bystander effects and adaptive responses--the Yin and Yang of low dose radiobiology? AB - Our current knowledge of the mechanisms underlying the induction of bystander effects by low doses of high or low LET ionizing radiation is reviewed. The question of what actually constitutes a protective effect is discussed in the context of adaptive (often referred to as hormetic or protective) responses. Finally the review considers critically, how bystander effects may be related to observed adaptive responses or other seemingly protective effects of low doses exposures. Bystander effects induce responses at the tissue level, which are similar to generalized stress responses. Most of the work involving low LET radiation exposure discussed in the existing literature measures a death response. Since many cell populations carry damaged cells without being exposed to radiation (so-called "background damage"), it is possible that low doses exposures cause removal of cells carrying potentially problematic lesions, prior to exposure to radiation. This mechanism could lead to the production of "U shaped" or hormetic dose-response curves. The level of adverse, adaptive or apparently beneficial response will be related to the background damage carried by the original cell population, the level of organization at which damage or harm are scored and the precise definition of "harm". This model may be important when attempting to predict the consequences of mixed exposures involving low doses of radiation and other environmental stressors. PMID- 15530546 TI - A biological-based model that links genomic instability, bystander effects, and adaptive response. AB - This paper links genomic instability, bystander effects, and adaptive response in mammalian cell communities via a novel biological-based, dose-response model called NEOTRANS3. The model is an extension of the NEOTRANS2 model that addressed stochastic effects (genomic instability, mutations, and neoplastic transformation) associated with brief exposure to low radiation doses. With both models, ionizing radiation produces DNA damage in cells that can be associated with varying degrees of genomic instability. Cells with persistent problematic instability (PPI) are mutants that arise via misrepair of DNA damage. Progeny of PPI cells also have PPI and can undergo spontaneous neoplastic transformation. Unlike NEOTRANS2, with NEOTRANS3 newly induced mutant PPI cells and their neoplastically transformed progeny can be suppressed via our previously introduced protective apoptosis-mediated (PAM) process, which can be activated by low linear energy transfer (LET) radiation. However, with NEOTRANS3 (which like NEOTRANS2 involves cross-talk between nongenomically compromised [e.g., nontransformed, nonmutants] and genomically compromised [e.g., mutants, transformants, etc.] cells), it is assumed that PAM is only activated over a relatively narrow, dose-rate-dependent interval (D(PAM),D(off)); where D(PAM) is a small stochastic activation threshold, and D(off) is the stochastic dose above which PAM does not occur. PAM cooperates with activated normal DNA repair and with activated normal apoptosis in guarding against genomic instability. Normal repair involves both error-free repair and misrepair components. Normal apoptosis and the error-free component of normal repair protect mammals by preventing the occurrence of mutant cells. PAM selectively removes mutant cells arising via the misrepair component of normal repair, selectively removes existing neoplastically transformed cells, and probably selectively removes other genomically compromised cells when it is activated. PAM likely involves multiple pathways to apoptosis, with the selected pathway depending on the type of cell to be removed, its cellular environment, and on the nature of the genomic damage. PMID- 15530547 TI - Efficacy of melatonin as protectant against oxidative stress and structural changes in liver tissue in pinealectomized rats. AB - Previous observations demonstrated that physiological levels of melatonin, the pineal secretory product, are important in protecting against oxidative stress induced tissue damage. We investigated the effects of pinealectomy and administration of exogenous melatonin on liver tissue in rats. Pinealectomized (Px) and sham-operated (non-Px) rats were used. We evaluated structural changes, reduced glutathione (GSH) levels and malondialdehyde (MDA) levels. Rats were divided into three groups (10 rats in each group): control (non-Px), Px+vehicle and Px+melatonin (4 mg/kg given daily intraperitoneally for 10 days). Liver GSH levels were significantly lower in Px rats than in the control group. Melatonin administration significantly increased GSH levels (p < 0.05). Px caused a significant increase in MDA levels as compared with the control group and melatonin administration to Px rats significantly reduced MDA levels in the liver (p < 0.05). Sinusoidal dilatation to a varying degree developed in all Px rats. Severity of mononuclear cell infiltration and sinusoidal congestion were lower in Px+melatonin group than in the Px group. These findings suggest that a significant increase in oxidative and structural changes occur in rat livers after pinealectomy, which can be diminished by melatonin treatment. PMID- 15530548 TI - The development of myelinated nociceptors is dependent upon trks in the trigeminal ganglion. AB - Cell size of primary sensory neurons and distribution patterns of neurons that are immunopositive (ip) for VRL-1, a newly cloned capsaicin-receptor homologue, were examined in trigeminal ganglia (TGs) of knockout mice for trkA, trkB or trkC to determine the developmental dependency of myelinated nociceptors on expression of the genes. The number of TG neurons was strongly decreased in the knockout mice as compared to wildtype and heterozygous mice (82%, 39%, and 48% reduction for trkA, trkB and trkC, respectively). The absence of trkA and trkC reduced the number of TG neurons in all cell-size ranges. The number of medium-sized and large TG neurons was decreased in trkB-knockout mice, whereas that of small TG neurons was barely affected by trkB deficiency. TG contained abundant VRL-1-ip neurons in wildtype and heterozygous mice; 9% of TG neurons exhibited immunopositivity. In trkA-knockout mice, VRL-1-ip neurons almost disappeared (1% of TG neurons were VRL-1-ip). However, 13% and 9% of TG neurons in trkB- and trkC knockout mice, respectively, were immunostained for the ion channel protein. In trkC-knockout mice, the proportion of large VRL-1-ip neurons decreased whereas that of small and medium-sized VRL-1-ip neurons increased. In addition, immunohistochemistry of the protein gene product 9.5 (PGP 9.5) demonstrated that trkA deficiency caused a marked reduction of varicose endings in the epithelium of the palatal mucosa. Loss of trkC diminished the number of PGP 9.5-ip varicose fibers in the deep layer of mucosal connective tissue of the palate. In tooth pulp, PGP 9.5-ip nerve fibers were absent in trkA-knockout mice but abundant in trkB- and trkC-knockout mice. The present study suggests that the development of myelinated nociceptors is dependent on trkA and trkC but not on trkB. PMID- 15530549 TI - Progesterone treatment and the progress of early pregnancy reduce desmoglein 1&2 staining along the lateral plasma membrane in rat uterine epithelial cells. AB - Uterine epithelium undergoes dramatic changes during early pregnancy in preparation for implantation. We have studied distribution patterns of the desmosomal marker, desmoglein 1&2, in rat uterine epithelial cells during early pregnancy as well as in hormonally stimulated ovariectomised animals. On day 1 of pregnancy as well as in oestradiol treated rats, desmoglein 1&2 staining was localized along the entire length of the lateral plasma membrane. By day 3 and on subsequent days of pregnancy as well as in ovariectomised animals treated with progesterone alone or in combination with oestradiol, desmoglein 1&2 staining was concentrated at the apical portion of the lateral plasma membrane. We suggest that the reorganisation of these desmosomal cadherins is an important component of uterine epithelial receptivity and this relocation is under the control of the ovarian hormone progesterone. PMID- 15530550 TI - Distribution patterns of matrix metalloproteinase (MMP)-2 and -9 and their inhibitors (TIMP-1 and TIMP-2) in the human decidua during early pregnancy. AB - Early human trophoblast shows dramatic invasive properties during early pregnancy. A tightly-regulated activation of matrix metalloproteinases (MMPs) is considered to be of critical importance for the control of trophoblast invasion. The aim of the present study was to determine MMP-2, MMP-9, TIMP-1 and TIMP-2 protein expression in decidual endometrium during the first trimester of pregnancy (22-42 days post coitus) with special attention to their expression patterns in endometrial compartments. Cytokeratin staining applied to adjacent sections was used to identify epithelial and trophoblast cells. We observed that MMP-2, particularly in the fourth week, appeared to be expressed more strongly in extravillous trophoblasts (EVTs) and vascular endothelial cells in the first trimester of pregnancy. Therefore, MMP-2 is likely to be the primary mediator in invasion of the trophoblast into the decidual endometrium, as well as in vascular remodeling and angiogenesis in the first trimester of pregnancy. The high expression of TIMP-1 and TIMP-2 in EVTs and glandular epithelium suggests that a restricted and balanced expression of these molecules is important for matrix remodeling and controlled trophoblast invasion during placentation. We conclude that (1) MMP-2 and MMP-9 and their inhibitors TIMP-1, and TIMP-2 determine the invasive behavior of trophoblast into the endometrium, and in particular, (2) MMP 2 may be the key regulator of trophoblast invasion in early human pregnancy. PMID- 15530551 TI - Ultrastructural and immunohistochemical similarities of two distinct entities; multiple sclerosis and hereditary motor sensory neuropathy. AB - In the present study, we present the ultrastructural and immunohistochemical properties of the sural nerves of two patients, one of whom was diagnosed as having multiple sclerosis with involvement of the peripheral nervous system (PNS), and the other as having hereditary motor sensory neuropathy type-I with involvement of the central nervous system (CNS). Expression of several extracellular matrix (ECM) proteins (fibronectin, laminin, and collagen type-IV), intermediate filaments (vimentin) and S-100 protein (marker for the axon-Schwann cell interface) was investigated by means of immunohistochemical methods. In addition, the tissue samples were evaluated ultrastructurally. Immunohistochemical staining revealed increased expression of the ECM molecules mentioned above in relation with the sural nerves of the patients. We hypothesize that this enhanced expression is due to Schwann cell-axon interactions. Vimentin expression was different in Schwann cells and S-100 immunostaining was decreased near the Schwann cell-axon interface. Myelin fragmentation, axon vacuolization, onion bulbs, tomoculous formation, axonal degeneration were found to occur. These results suggest that there is active ECM reorganization in the sural nerve of these patients, and some ultrastructural changes are similar in the damaged axonal organization and in Schwann cells although the changes are not completely the same in the two patients. In conclusion, our study demonstrates that there is an association between the demyelinization process in the CNS and the PNS even though they are affected by different mechanisms. PMID- 15530552 TI - Evidence for the presence of the sphingosine-1-phosphate receptor Edg-8 in human radial glial fibers. AB - The distribution pattern of Edg-8 immunostaining in the human developing brain has been investigated with special reference to radial glial fibers. At 24 weeks of gestation, fragments of radial glial fibers are Edg-8-positive within the cortical plate and subplate of allocortical areas. These Edg-8-positive fragments often appear enlarged as varicosities and some of them terminate at blood vessels. Between 28 and 30 weeks of gestation, all iso- and allocortical areas contain Edg-8-immunolabelled radial glial fibers revealing curvature next to sulci. After 32 weeks of gestation, radial glial fibers gradually disappear; instead Edg-8-positive transitional stages between radial glia and astrocytes were found. The findings indicate that sphingosine-1-phosphate may play a regulatory role in the transformation of radial glial cells into astrocytes and may affect proliferative activity of these cells. PMID- 15530553 TI - Differentiation of the epidermis in turtle: an immunocytochemical, autoradiographic and electrophoretic analysis. AB - Proteins involved in the process of cornification of turtle epidermis are not well known. The present immunocytochemical, electrophoretic and autoradiographic study reports on the localization patterns and molecular weights of keratins, which are cornification proteins, and of tritiated histidine in turtle epidermis. Alpha-keratins with a molecular weight of 40-62 kDa are present in the epidermis. Beta-keratin is mainly detectable in the stratum corneum of the carapace and plastron, but is rarely present or even absent in the corneous layer of limb, tail and neck epidermis. After electrophoresis and immunoblotting with an antibody against chicken scale beta-keratin, bands at 15-17, 22-24, and 36-38 kDa appeared. This antibody recognized weaker bands at 38-40 and 58-60 kDa in the soft epidermis. After reduction and carboxymethylation of proteins extracted from carapace and plastron, but not of proteins from the soft epidermis, protein bands at 15-17 and 35-37 kDa were found when using the anti-beta 1-keratin antibody. Loricrin-, filaggrin-, sciellin-, and transglutaminase-like immunostaining was detectable only in the transitional and lowermost corneous layers of the soft epidermis. Vesicular bodies in the transitional layer were immunolabeled by the anti-loricrin antibody, and weakly by the anti-filaggrin and anti transglutaminase antibodies. In immunoblots, the anti-loricrin antibody reacted with a major band at 50-54 kDa in both carapace-plastron and soft epidermis. The anti-sciellin antibody detected major bands at 38-40 and 50 kDa in hard epidermis, and at 50 and 54-56 kDa in soft epidermis. Filaggrin-like immunostained bands were observed at 50-55 and 62-64 kDa. This immunostaining was probably due to a common epitope in filaggrin and some keratins. Histidine was evenly incorporated in the epidermis, and the ultrastructural study showed random labeling, often associated with keratin bundles of alpha and beta-keratinocytes. Histidine-labeled protein bands were not found in the carapace-plastron. In the soft epidermis, weakly labeled bands at 15-20, 25, and 45-60 kDa were found occasionally. The latter bands probably represented neo-synthesized keratins as was also indicated by the ultrastructural autoradiographic analysis. In conclusion, our study suggests that proteins with epitopes that they have in common with cornification proteins of mammalian epidermis are also present in the epidermis of turtle. PMID- 15530554 TI - An analysis of a wrong Nobel Prize-Johannes Fibiger, 1926: a study in the Nobel archives. PMID- 15530555 TI - Tumor-host immune interactions and dendritic cell dysfunction. AB - Several lines of evidence from recent years support the existence of cancer immunosurveillance, especially studies of natural killer (NK) cells and the IFN gamma pathway. However, immune suppression is clearly observed in cancer patients and tumor-bearing animals as well. The fact is that although cancers often elicit a vigorous immune response during the early part of their growth, the immune response is soon down-regulated, permitting progressive tumor growth. Apparently, the intrinsic plasticity of tumors allows the immune system to sculpt the immunogenic phenotypes of tumors to escape efficient immune destruction. But most evidently, several mechanisms have now been found to contribute to the failure of immune control of tumor growth. Tumor cells have a very low level of MHC class II, costimulatory molecules, and weak antigens. They also produce immune suppressive factors (VEGF, IL-10, PGE(2)) that exert systemic effects on immune cell function. In particular, disabled dendritic cell differentiation, maturation, migration, and function are fundamental to this defect, as they are the most potent antigen-presenting cells (APCs) of the immune system, interacting with T and B lymphocyte as well as NK cells to induce and modulate immune responses. In addition, tumors also alter host hematopoiesis and produce large numbers of immature dendritic cells, and evidence shows that these cells are directly immune suppressive. Harnessing the immune system for effective cancer therapy has remained a great challenge. DC-based vaccines, or DC-based vaccines in combination with treatments designed to improve the host immune environment, may offer hope for more effective cancer immunotherapy. Tumor-host interactions are an important determinant of tumor behavior and response to therapy. How tumors interact with their hosts is thus a very broad and complex topic. In this chapter, we will focus on tumor-host immune interactions and the roles of dendritic cell dysfunction in tumor avoidance of host immune responses. We will survey recent findings regarding tumor immune surveillance, antitumor host immune responses, and how the immune system also functions to promote or select tumor variants with reduced immunogenicity. We will then discuss immune suppression caused by tumors, which is clearly observed in tumor-bearing animals and cancer patients. Finally, we will discuss altered dendritic cell function and differentiation in some detail, as it is likely to be one of the most fundamental mechanisms by which tumors escape immune responses. PMID- 15530556 TI - Noninvasive imaging of reporter gene expression in living subjects. AB - The development of noninvasive imaging technologies designed specifically for use with small animals has provided new paradigms for cancer research. Traditional molecular biology techniques are being melded with noninvasive imaging technologies to develop a new research domain, "molecular imaging." One of the most exciting advances in this research area is the adaptation and application of conventional reporter-gene imaging techniques, used extensively by cell and molecular biologists, to living animals. Using these new assays, investigators can image noninvasively, repeatedly, and quantitatively the location, magnitude, and duration of reporter-gene expression in living animals. This review will describe the instrumentation used for noninvasive imaging of reporter genes, the reporter genes developed for noninvasive imaging with radio-nuclide-based assays such as positron emission tomography, and the reporter genes used for optically based noninvasive assays using sensitive charged-coupled device cameras. Applications of noninvasive, whole-animal imaging to gene therapy for cancer, to cell-based therapy for cancer, to lymphocyte activation, to cancer progression and dissemination in engrafted models, to tumor initiation, promotion and metastasis in conditional murine models of cancer induction, and to the noninvasive monitoring of tumor responses to a variety of therapies are described. New developments in multimodality molecular imaging are discussed, and the potential utility of noninvasive reporter gene expression in the diagnosis and management of human cancer is presented. PMID- 15530557 TI - A role for Id proteins in mammary gland physiology and tumorigenesis. AB - Id helix-loop-helix (HLH) proteins are regulators of cell growth and differentiation in embryonic and adult tissues. They are members of the basic HLH family of transcription factors but lack a DNA binding domain. By binding to basic HLH transcription factors, Id proteins regulate gene expression. Id1 and Id3 have extensive sequence homology and similar patterns of expression during embryogenesis and in adult tissues. They are also expressed at high levels in the endothelial cells of tumor-infiltrating blood vessels, and breast tumors spontaneously arising in MMTV-neu mice demonstrate impaired angiogenesis when growing in an Id1- and/or Id3-deficient background. These lesions are typically cystic with a small rim of viable tumor cells surrounding an acellular necrotic core. Id2 plays a critical role in breast differentiation and lactation. Id4 regulates BRCA1 expression and may be involved in hormone-dependent regulation of BRCA1 homeostasis. Thus, all four members of the Id protein family play pivotal roles in distinct aspects of normal and malignant breast biology, the subject of this review. PMID- 15530558 TI - p38 MAP kinase's emerging role as a tumor suppressor. AB - The p38 proteins are an evolutionally conserved family of mitogen-activated protein kinases (MAPK). Recent studies have led to progress in our understanding the roles of p38 MAPK in regulation of tumorigenesis through key cellular growth control mechanisms. Along with the previously well-characterized proapoptotic functions, new data highlight the critical contributions of p38 MAPK in the negative regulation of cell cycle progression. This review will focus on the ability of p38 MAPK to positively regulate several tumor suppressor (p53- and Rb dependent) pathways and to attenuate oncogenic (Cdc25A and Cdc25B phosphatases) signals. The concept of p38 MAPK as a potential tumor suppressor will be developed. PMID- 15530559 TI - Deregulation of the cell cycle by the Epstein-Barr virus. PMID- 15530560 TI - Duck hepatitis B virus: an invaluable model system for HBV infection. PMID- 15530561 TI - Novel insights into hepatitis C virus replication and persistence. AB - Hepatitis C virus (HCV) is a small enveloped RNA virus that belongs to the family Flaviviridae. A hallmark of HCV is its high propensity to establish a persistent infection that in many cases leads to chronic liver disease. Molecular studies of the virus became possible with the first successful cloning of its genome in 1989. Since then, the genomic organization has been delineated, and viral proteins have been studied in some detail. In 1999, an efficient cell culture system became available that recapitulates the intracellular part of the HCV life cycle, thereby allowing detailed molecular studies of various aspects of viral RNA replication and persistence. This chapter attempts to summarize the current state of knowledge in these most actively worked on fields of HCV research. PMID- 15530562 TI - The regulation and maturation of antiviral immune responses. PMID- 15530563 TI - Prospects for the therapy and prevention of dengue virus infections. PMID- 15530564 TI - Bacteriophage T4: structure, assembly, and initiation infection studied in three dimensions. PMID- 15530565 TI - Genomic organization, biology, and diagnosis of Taura syndrome virus and yellowhead virus of penaeid shrimp. PMID- 15530566 TI - Viruses of the chestnut blight fungus, Cryphonectria parasitica. PMID- 15530567 TI - GABAA receptor structure-function studies: a reexamination in light of new acetylcholine receptor structures. PMID- 15530568 TI - Dopamine mechanisms and cocaine reward. PMID- 15530569 TI - Proteolytic dysfunction in neurodegenerative disorders. PMID- 15530570 TI - Neuroimaging studies in bipolar children and adolescents. PMID- 15530571 TI - Chemosensory G-protein-coupled receptor signaling in the brain. PMID- 15530572 TI - Disturbances of emotion regulation after focal brain lesions. PMID- 15530573 TI - The use of Caenorhabditis elegans in molecular neuropharmacology. PMID- 15530574 TI - Correlates of weight loss and muscle-gaining behavior in 10- to 14-year-old males and females. AB - BACKGROUND: This study examined the influence of appearance and social acceptance esteem, awareness and internalization of media stereotypes, body size acceptance, and teasing on the weight loss and muscle-gaining behaviors of 10- to 14-year-old boys and girls. METHODS: Male (n = 670) and female (n = 788) students were drawn from one of four public senior middle schools (grades 6-8) in Southern Ontario as part of a longitudinal outcome-based study. Students' baseline self-report questionnaires, measuring the above variables, were analyzed for the purposes of this study. RESULTS: A higher percentage of girls reported engaging in weight loss behaviors, whereas a higher percentage of boys admitted to muscle gaining and the use of specific weight control methods such as laxative use and vomiting. Regression analyses revealed that internalization of media messages and body size acceptance were equally predictive of boys' weight loss and muscle-gaining behaviors, while teasing was found to also predict their muscle-gaining behavior. Among the girls, appearance esteem, internalization of media stereotypes, and body size acceptance were predictive of weight loss behaviors. None of the study variables were predictive of girls' muscle-gaining behavior. CONCLUSIONS: Weight loss and muscle-gaining behaviors appear to have their onset in children as young as 10 years. The findings support the need for prevention programs that focus on media literacy and ways to decrease weight-based teasing in the school setting. PMID- 15530575 TI - Expired air carbon monoxide concentration in mothers and their spouses above 5 ppm is associated with decreased fetal growth. AB - BACKGROUND: Smoking during pregnancy is associated with reduced birthweight; this relation can be reversed by smoking cessation. Some but not all previous studies have shown that smoking reduction (measured as cigarettes/day or urinary cotinine) may also improve birthweight. The relationship between maternal and spouses' expired air carbon monoxide (CO) concentrations (EACO) on fetal growth has not yet been evaluated. METHODS: Eight hundred fifty-six smoking and nonsmoking pregnant women were followed during their pregnancy. Their EACO was determined in the first trimester and during delivery. The spouses' EACO were also measured at delivery. The main outcome measure was the infants' birthweight. Secondary measures included head circumference, Apgar score, and heart rate at delivery. Cord blood fetal carboxyhemoglobin (FCOHb) served as internal control. RESULTS: Birthweight dose-dependently and significantly decreased with increasing level of maternal (0-5: 3406 +/- 32; 6-10: 3048 +/- 57; 11-20: 2858 +/- 54; >20 ppm: 2739 +/- 34 g, P < 0.0001) or spouses' EACO (0-5: 3546 +/-25; 6-10: 3484 +/- 51; 11-20: 3309 +/- 47; >20 ppm: 3190 +/- 57 g, P < 0.0001). Even the birthweight of newborns whose mother had EACO between 6 and 10 ppm was significantly lower than the birthweight of newborns whose mother had an EACO between 0 and 5 ppm. Spouses' EACO of delivering women with EACO of 0-5 ppm showed similar effect. Head circumference, Apgar score, and normal term gestational age decreased also significantly with increasing maternal or spouses' EACO. CONCLUSIONS: Both maternal and spouses' EACO measured during delivery, a proxy of EACO during pregnancy, were dose-dependently and inversely associated with fetal growth. Even low maternal (6 to 10 ppm) or spouses' (11 to 20 ppm) EACO may be associated with significantly lower birthweight. PMID- 15530576 TI - Tobacco advertising in communities: associations with race and class. AB - BACKGROUND: Individuals of lower socioeconomic position smoke at higher rates than those of higher socioeconomic position. Because of this disparity, the National Cancer Institute has called for studies of targeted tobacco marketing to clarify mechanisms contributing to higher tobacco use among low-income Americans and other high-risk populations. METHODS: We observed tobacco industry marketing in six Boston area communities (two of high socioeconomic position and four of low position; total of 41 observations) and in selected print publications that circulated in those communities during a 22-month period in 2000-2002. RESULTS: On average, there were fewer tobacco advertisements in the higher socioeconomic communities, compared to the lower socioeconomic communities (P < 0.001). In the low socioeconomic communities, there were more than three times as many brand advertisements as youth access signs (P = 0.0012). Although brand advertisements outnumbered smoke-free signs, on average, there was no difference in the ratio of brand advertisements to smoke-free signs in low and high socioeconomic communities (P = 0.06). CONCLUSIONS: The tobacco industry is actively present in community settings, particularly in communities with a low socioeconomic profile (SEP). Tobacco control researchers and advocates need to continue to monitor the tobacco industry's behavior at the community level and develop strategies to counter this behavior. PMID- 15530577 TI - The neurocognitive effects of alcohol on adolescents and college students. AB - BACKGROUND: Adolescents and college students are at high risk for initiating alcohol use and high-risk (or binge) drinking. There is a growing body of literature on neurotoxic and harmful cognitive effects of drinking by young people. On average, youths take their first drink at age 12 years. METHODS: MEDLINE search on neurologic and cognitive effects of underage drinking. RESULTS: Problematic alcohol consumption is not a benign condition that resolves with age. Individuals who first use alcohol before age 14 years are at increased risk of developing alcohol use disorders. Underage drinkers are susceptible to immediate consequences of alcohol use, including blackouts, hangovers, and alcohol poisoning and are at elevated risk of neurodegeneration (particularly in regions of the brain responsible for learning and memory), impairments in functional brain activity, and the appearance of neurocognitive deficits. Heavy episodic or binge drinking impairs study habits and erodes the development of transitional skills to adulthood. CONCLUSIONS: Underage alcohol use is associated with brain damage and neurocognitive deficits, with implications for learning and intellectual development. Impaired intellectual development may continue to affect individuals into adulthood. It is imperative for policymakers and organized medicine to address the problem of underage drinking. PMID- 15530578 TI - Factors associated with physical activity among older people--a population-based study. AB - BACKGROUND: Regular physical activity improves survival and functional ability, and can improve quality of life. Few studies have examined factors associated with physical activity in older adults at a population level. This study examines factors associated with physical activity in the New South Wales (NSW) older population. METHODS: Data from a random survey of 8881 community-dwelling people aged 65+ years were examined. We used Cox's regression to analyze the influence of demographic, health, and social factors on physical activity. 'Adequate' physical activity was defined as at least 30 min of walking, moderate or vigorous activity on at least 5 days in the last week. Reported barriers to physical activity were also examined. RESULTS: Adequate physical activity was independently associated with sex (male), younger age, ability to travel independently, better physical functioning, lower psychological distress, rural residence, not having diabetes, adequate fruit and vegetable intake, and speaking a language other than English at home. No independent associations were found for good long distance eyesight, being a caregiver, living alone, age left school, employment status, fear of falling, or perception of neighborhood safety. Health problems were frequently reported barriers to physical activity. CONCLUSIONS: About half of older adults report adequate physical activity. These results will inform strategies to promote physical activity among older people. PMID- 15530579 TI - Patient knows best: blinded assessment of nonadherence with antituberculous therapy by physicians, nurses, and patients compared with urine drug levels. AB - BACKGROUND: Adherence with therapy is a wide spectrum of behavior rather than a categorical state. While extreme nonadherence is readily apparent, it is rare compared to lesser degrees of nonadherence, which are difficult to predict. AIMS: To compare the accuracy of doctor, nurse, and patient prediction of adherence with antituberculous therapy with urine isoniazid levels. METHODS: A prospective, blinded clinical study was conducted, comparing adherence to antituberculous therapy as reported by patients, doctors, and nurses with urine isoniazid levels. We studied 173 patients with active tuberculosis (TB) recruited over 3 years in two TB clinics in Victoria, Australia. Adherence was defined as six random urine isoniazid (INH) levels being >0. Blinded assessment of adherence was completed by doctors, nurses, and patients. Lid opening and closing of computerized pill bottles were measured in a random subsample. RESULTS: Of 173 patients, the rate of nonadherence was 24% (41/173) by urine INH, 54% (93/173) by patient self report, 11% (19/173) by doctor assessment, and 7% (12/173) by nurse assessment. The sensitivity of prediction of nonadherence was 76% for patient self-report, 24% for doctor assessment, and 19% for nurse assessment. The 10 patients who used computerized pill bottles were all (100%) noncompliant at some stage. CONCLUSION: Nonadherence is common and poorly predicted by doctors and nurses, even those with extensive experience in treating TB. Contrary to popular belief, patient self-report is more reliable than doctor or nurse assessment of nonadherence. As clinicians, asking patients about adherence may be more valuable than attempting to judge for ourselves. PMID- 15530580 TI - Use of complementary and alternative medicine among United States adults: the influences of personality, coping strategies, and social support. AB - BACKGROUND: Although patterns of utilization of complementary and alternative medicine (CAM) in the community have begun to be described, few studies have addressed the relationships between dispositional psychological factors and the use of CAM. The aim of this study was to examine the associations between CAM use and personality, coping strategies, and perceived social support in a representative sample of adults in the United States. METHODS: Data were drawn from the Midlife Development in the United States Survey (MIDUS), a representative sample of 3,032 adults aged 25-74 in the US population. We analyzed use of acupuncture, biofeedback, chiropractic, energy healing, exercise/movement therapy, herbal medicine, high-dose megavitamins, homeopathy, hypnosis, imagery techniques, massage, prayer/spiritual practice, relaxation/mediation, and special diet within the last year. Multiple logistic regression analyses were used to evaluate the association of personality, dispositional coping strategies (primary and secondary control), and perceived social support and strain with CAM use, controlling for sociodemographic factors, medical care access, and physical and mental disorders. RESULTS: Openness was positively associated with the use of all types of CAM except manipulative body based methods. Extroversion was inversely correlated with the use of mind-body therapies. Primary control was inversely and secondary control directly correlated with the use of CAM. Perceived friend support was positively associated with the use of mind-body therapies, manipulative body-based methods, and alternative medical systems. Perceived partner strain was positively associated with the use of biologically based therapies, and family strain increased the odds of manipulative body-based methods. CONCLUSIONS: This study is the first to document a significant association between specific domains of personality, coping strategies, and social support, and the use of CAM among adults in the general population. Understanding the relationships between psychological factors and CAM use may help researchers and health care providers to address patients' needs more effectively and to achieve better adherence to treatment recommendations. PMID- 15530581 TI - Engagement and retention of participants in a physical activity website. AB - BACKGROUND: Websites have the potential to deliver enhanced versions of targeted and tailored physical activity programs to large numbers of participants. We describe participant engagement and retention with a stage-based physical activity website in a workplace setting. METHODS: We analyzed data from participants in the website condition of a randomized trial designed to test the efficacy of a print- vs. website-delivered intervention. They received four stage targeted e-mails over 8 weeks, with hyperlinks to the website. Both objective and self-reported website use data were collected and analyzed. RESULTS: Overall, 327 were randomized to the website condition and 250 (76%) completed the follow-up survey. Forty-six percent (n = 152) visited the website over the trial period. A total of 4,114 hits to the website were recorded. Participants who entered the site spent on average 9 min per visit and viewed 18 pages. Website use declined over time; 77% of all visits followed the first e-mail. CONCLUSIONS: Limited website engagement, despite the perceived usefulness of the materials, demonstrates possible constraints on the use of e-mails and websites in delivering health behavior change programs. In the often-cluttered information environment of workplaces, issues of engagement and retention in website delivered programs require attention. PMID- 15530582 TI - A breast-feeding promotion and support program a randomized trial in The Netherlands. AB - BACKGROUND: In the Netherlands, the initiation rate of breast-feeding (BF) was 80% in 2002, but only 35% of the mothers continued to breast-feed for 3 months. This study examined the effectiveness of a breast-feeding promotion program to increase the continuation of breast-feeding. METHODS: A cluster-randomized intervention trial was used. Ten child health care centers in three regions of the home health care were randomly allocated to the program or usual care. Elements in the program were health counseling, measures to enhance cooperation, early signaling of breast-feeding problems and continuity of care, and lactation consultancy. Pregnant mothers who applied for home health care in the intervention or usual care regions were enrolled and were followed up from pregnancy until 6 months postpartum (n = 683). The primary outcome measure was the continuation of breast-feeding until at least 3 months. RESULTS: The 3-month breast-feeding rate was 32% in the intervention and 38% in the control groups (OR = 0.79, 95% CI = 0.58-1.08). CONCLUSION: The program was not effective. We discuss possible explanations from the design and execution of the trial and give some points for improvement of our program, such as the categories of caregivers involved and the number and duration of contacts after parturition. PMID- 15530583 TI - Predictors of weight control advice in primary care practices: patient health and psychosocial characteristics. AB - BACKGROUND: Past research has surveyed primary care physicians (PCP) about their attitudes and practices towards obese patients, yet less is known about the patients receiving advice. METHODS: The Primary Care Weight Control Project (PCWC) enrolled 18 PCPs in a randomized clinical trial and asked 255 of their patients who were either overweight or obese at baseline about past weight control advice. RESULTS: At baseline, 66.4% of patients reported that their physician previously told them they were overweight. Body mass index (BMI) was a strong predictor of being identified as obese. While 65.1% received information on the health benefits of weight loss, only 36.6% of patients were ever given specific weight control advice, and 28.2% were advised to increase their physical activity. A history of type 2 diabetes, high cholesterol, or hypertension was associated with physician-delivered weight control advice. Stages of change and number of prior visits with the PCP were also associated with physician advice. CONCLUSIONS: Patients were more likely to receive education about weight loss than specific behavioral advice on how to lose weight. Physicians were more likely to provide weight control advice to their patients who had obesity-related comorbidities than to patients who were overweight or obese and without risk factors. PMID- 15530585 TI - The effects of short- vs. long-bout exercise on mood, VO2max, and percent body fat. AB - BACKGROUND: To compare the ACSM-CDC physical activity accumulation recommendation to the traditional recommendation, for impact on mood and physiological markers of fitness. METHODS: Randomized controlled trial with sedentary male (n = 21) and female (n = 19) subjects assigned to walk either long bouts (LB; 30 min/day), short bouts (SB; 3 x 10 min/day), or a nonexercise control (CTL) group for 8 weeks. Pre- and post-measures were collected for V02max and percent body fat. Pre , mid-, and post-measures were collected for the Profile of Mood States (POMS). RESULTS: VO2max increased in the SB group (+7.2%) and LB (+6.7%; P < or = 0.05). Percent body fat decreased in the LB group (-6.7%; P < or = 0.05). Total mood disturbance (TMD) decreased in the LB and SB groups (P < or = 0.05); only the LB group showed reductions compared to the CTL group (P < or = 0.05). Tension anxiety and vigor-activity were altered in the LB group compared to the other two groups (P < or = 0.05). Reductions in percent body fat correlated with TMD (r = 0.38; P < or = 0.05) and Tension-anxiety reduction (r = 0.40; P < or = 0.05). CONCLUSIONS: LB and SB walking produced similar and significant improvements in VO2max LB walking was more effective at reducing percent body fat, tension anxiety and total mood disturbance, and increasing vigor compared to the control group. PMID- 15530584 TI - Family characteristics and smoking among urban and rural adolescents living in China. AB - BACKGROUND: Adolescent smoking is a public health concern in China. Although the family is an important social influence in China, few associations among family characteristics and adolescent smoking have been examined using Chinese samples. METHODS: Survey data on psychosocial variables and smoking were collected from a sample of 3629 7th grade adolescents (46% female; 54% male; mean age 12.7 years) in Wuhan, China. For adolescents, past 30-day smoking, family relationships, parents' negative sanctioning of smoking, parents' agreement with smoking, and parents' smoking behaviors are assessed. To account for the clustered data structure, hierarchical logistic regression analyses controlling for demographics (urbanization, age) examined the independent and multivariate effects of family characteristics for each gender. RESULTS: Girls are less likely than boys to report smoking and are more likely to report positive family relationships, and having parents with negative attitudes toward them smoking. Positive family relationships and age were strongly associated with smoking for both genders. No significant differences exist by gender. CONCLUSION: These findings suggest that the quality of family relationships are important for adolescent female and male smoking in China. PMID- 15530586 TI - Number of children and the risk of obesity in older women. AB - BACKGROUND: The purpose of this study is to examine number of live births, other reproductive factors, and health behaviors in relation to obesity risk among older women. METHODS: Data were collected during in-person and telephone-based interviews from a population-based cohort in Utah. RESULTS: A total of 2,035 women aged 66-102 are included in this report. Overall, 403 (20%) older women were determined to be obese. The rates of obesity were significantly higher with increasing numbers of children, demonstrating a dose-response relationship (P < 0.05). After adjustment for age, education, marital status, BMI at age 18, use of oral contraceptives, hysterectomy status, physical activity, current use of hormone therapy, and age at menarche, the risk of obesity increased 11% with each additional live birth. In additional analyses that excluded nulliparous women, after adjusting for cumulative months of breast-feeding, the risk of obesity increased 7% with each live birth. In this cohort of older women, we found higher rates of obesity with increasing number of children that was independent of socioeconomic status and other confounding factors. CONCLUSIONS: In a cohort of older women, higher rates of obesity were associated with increasing number of children that was independent of socioeconomic status and other confounding factors. PMID- 15530587 TI - Dietary intake among young adolescents in Ontario: associations with vegetarian status and attitude toward health. AB - BACKGROUND: This study aimed to describe the dietary intakes of adolescent vegetarians and omnivores and determine if, and how, attitude toward personal health related to food consumption. METHODS: Among grade 9 students in Ontario, Canada (n = 630), vegetarian status and estimated consumption of foods and food groups were determined by food frequency questionnaire. Personal health was self categorized as very important (the "health conscious") or somewhat/not important (the "non-health conscious"). RESULTS: The prevalence of vegetarianism was 6.5% (CI = 4.6-8.4%) among females and 1.0% (CI = 0.2-1.8%) among males. Health conscious omnivores consumed more grain, vegetables and fruit, and milk product than non-health-conscious omnivores (P < 0.05). Health-conscious vegetarians ingested more grain products, vegetables and fruit, and meat and alternatives than non-health-conscious vegetarians (P < 0.05). Among non-health-conscious vegetarians, none consumed two daily servings of meat and alternatives compared to 60.5% of non-health-conscious omnivores (P < 0.001). Among health-conscious vegetarians, milk product consumption was lower than that of health-conscious omnivores (P = 0.015). CONCLUSIONS: Large proportions of both vegetarian and omnivore adolescents consumed suboptimal diets. Health consciousness had value as an indicator of dietary adequacy and may be useful as a rudimentary screen for problematic dietary consumption patterns. PMID- 15530588 TI - Smoking reduction intervention in a large population-based study. The Inter99 study. AB - BACKGROUND: Smoking reduction has been introduced as an alternative to smokers unable or unwilling to quit but has never been implemented in a population-based intervention. METHODS: Two thousand four hundred eight daily smokers in all motivational stages were included in a randomised population-based intervention study, in Copenhagen, Denmark. Smokers, unwilling or unable to quit, were encouraged to reduce their tobacco consumption. Furthermore, smokers in the high intensity intervention were offered participation in smoking reduction groups. RESULTS: Twenty-three percent of those who attended both baseline and 1 year visit reported reduction by at least 5 g and 8% reported a halving or more. Halving of tobacco consumption was achieved significantly more often than in the background population, OR = 2.6 (1.6-4.4), even when assuming that non participants had not reduced, OR = 1.7 (1.0-2.8). Reduction of at least 5 g doubled the probability of increased motivation to quit and a halving increased it more than four times. The reductions were not validated. Less than 2% attended the smoking reduction groups. CONCLUSION: The smoking reduction intervention was significant in self-reported reduction of tobacco consumption and subsequently increased motivation to quit. This may open new perspectives, with reduction as a first step towards cessation, a possible supplement to smoking cessation strategies. PMID- 15530589 TI - Psychosocial and environmental factors associated with physical activity among city dwellers in regional Queensland. AB - BACKGROUND: Research has recently adopted the use of social-ecological models in the study of physical activity. Few studies, however, have addressed the influence of the environment on activity using Geographic Information System (GIS)-derived measures of environmental attributes and self-report ratings of other environmental attributes. Even fewer have examined walking behaviors. METHODS: Self-report measures of physical activity, social support, self efficacy, and perceived neighborhood environment were obtained by means of a Computer-Assisted-Telephone-Interview (CATI) survey of 1,281 residents of Rockhampton, Queensland. Over 94% (1,215) of respondents' residential locations were successfully geocoded into the existing city council GIS database. The self report data, along with GIS-derived measures, were used to determine the relationships among selected variables of the neighborhood environment for each geocoded location. RESULTS: GIS-derived measures of street connectivity and proximity to parkland, the number of active people in a 1-km radius, and self reported perceptions of neighborhood cleanliness showed associations with the likelihood of achieving sufficient levels of physical activity when adjusting for selected psychosocial variables. GIS-derived Euclidian distance to footpath networks, number of dogs in 0.8-km radius, network distance to newsagents, and perceptions of footpath condition were significantly associated with the likelihood of participating in any recreational walking. CONCLUSION: Environmental characteristics were found to have differential influences on the two selected measures of physical activity. Aesthetics and safety appear to be important influences of physical activity, whereas proximal footpaths showed increased likelihood of participation in recreational walking. It is proposed that the strength of association between the environmental and physical activity may be improved if future research utilizes a Geographic Information System approach to the study of restricted geographical areas. PMID- 15530590 TI - A combined breast health/weight loss intervention for Black women. AB - BACKGROUND: Overall incidence of breast cancer is slightly lower, but mortality rates are higher, for Black women compared to White women. Higher body mass index (BMI), sedentary lifestyles, and lower compliance with recommended breast health behaviors may contribute to higher risk and mortality. METHODS: A randomized pilot intervention trial was conducted to assess feasibility and efficacy of a combined breast health/weight loss intervention for 64 overweight or obese Black women, ages 35-65. The primary objectives were to determine whether a 20-week (twice weekly) intervention could decrease weight and dietary fat intake and increase physical activity and breast self-exam (BSE) proficiency. RESULTS: The project was implemented in two cohorts and retention was high for both (96% and 89%, respectively). Both cohorts showed increased proficiency in BSE in the intervention versus the control group (2.4 vs. -0.4, P<0.05; 3.3 vs. -0.2, P<0.001, respectively), but only cohort 2 showed decreased percent body weight (4.0% decrease vs. 0.9% increase, P<0.01), increased physical activity frequency (2.4 vs. 0.1 times/week, P<0.05), and a trend for decreased dietary fat (-2.6% kcal vs. 0.0% kcal, P=0.07) in the intervention compared to the control group. CONCLUSION: Few studies have documented weight loss among Black women, and no combined breast health/weight loss intervention has been conducted. This study documents the feasibility of recruiting, randomizing, and retaining women in a combined intervention and demonstrated weight loss and associated lifestyle changes. PMID- 15530591 TI - Vaccinations in pneumonia (VIP): pneumococcal and influenza vaccination patterns among patients hospitalized for pneumonia. AB - BACKGROUND: Although the CDC ACIP (Advisory Committee on Immunization Practices) recommends that appropriate inpatients receive pneumococcal and influenza vaccines, adult vaccination rates for these remain low. We therefore examined perihospitalization vaccination rates for high-risk pneumonia inpatients. METHODS: A retrospective chart review of all pneumonia patients admitted to one community hospital from 6/1/95 to 5/31/96. Vaccination history, co-morbidity, mortality, and prior and subsequent pneumonia admissions were recorded. Primary care providers and nursing homes were contacted to complete and verify vaccine histories. RESULTS: For 173 total admissions (160 subjects), vaccine histories were documented in the hospital chart in less than 0.5% of patients. While 97% had indications for both vaccines at the time of admission, no vaccines were given in the hospital and less than 5% had documented vaccinations during the subsequent 3 years. CONCLUSIONS: Despite clear indications, few patients had documented vaccination at any time. These data lend urgency to the recommendation that pneumococcal and influenza vaccines should be routinely administered to pneumonia inpatients at discharge. Furthermore, they illustrate the need for an improved method for tracking individual adult vaccinations. PMID- 15530592 TI - Training nurses and social workers in smoking cessation counseling: a population needs assessment in Hong Kong. AB - BACKGROUND: To achieve greater coverage of elderly smokers and to shift entire populations toward cessation, the provider-client interface could be broadened beyond physicians to include nurses and social workers, who can be formally trained to provide such services. We carried out a population-based training needs assessment of the latter two groups in Hong Kong. METHODS: Three thousand seven hundred eligible hospital-based nurses and 2,258 social workers who had elderly clients in Hong Kong were recruited in a knowledge, attitude, and practice (KAP) cross-sectional survey. We used multivariable logistic regression to identify predictors for two key outcomes-"initiation and advice" (ask and advise) and "follow-through" (assess, assist and arrange), based on the U.S. Agency for Health Care Policy Research framework. RESULTS: One thousand eight hundred forty-three (49.8%) nurses and 1,499 (66.4%) social workers responded. Nurses reported a much higher level of engagement in smoking cessation activities than social workers in all five steps of the AHCPR framework (P<0.001). Nurses (mean score=2.99+/-0.40 on a 4-point Likert scale) had more positive attitudes to tobacco control and smoking cessation counseling compared to social workers (mean score=2.79+/-0.41; P<0.001), whereas the latter group perceived themselves as more competent in handling such practice (mean score(nurses)=2.36+/-0.52, mean score(social workers)=2.51+/-0.39; P<0.001). Both attitudinal and self-perceived competence scores predicted incremental gains in the likelihood of offering "follow-through" interventions in addition to those observed for "initiation and advice" actions. CONCLUSION: Our findings highlight a large degree of unmet need in Hong Kong's hospital-based nurses and social workers who work with the elderly regarding smoking cessation service provision and training. Future research should focus on developing and evaluating programs that encourage nurses and social workers to provide cessation interventions to exert a much greater collective impact than doctors can alone. PMID- 15530593 TI - What intensity of physical activity do previously sedentary middle-aged women select? Evidence of a coherent pattern from physiological, perceptual, and affective markers. AB - BACKGROUND: The intensity of physical activity has been found to be inversely related to adherence, thus contributing to the problem of physical inactivity. Although most physical activity is unsupervised and participants, therefore, self select the intensity, very little is known about the level of intensity that they select. We hypothesized that participants would select, on average, an intensity proximal to the level of transition from aerobic to anaerobic metabolism. METHODS: Twenty-three middle-aged, formerly sedentary women participated in (a) an incremental treadmill test to determine their maximal aerobic capacity and gas exchange ventilatory threshold, an index of the aerobic-anaerobic transition, and (b) a 20-min bout of treadmill exercise during which they were allowed to select the speed. RESULTS: On average, but with considerable interindividual variability, the women selected an intensity that, in terms of treadmill speed, heart rate, oxygen uptake, and perceived exertion was no different from the intensity corresponding to their gas exchange ventilatory threshold. Moreover, affective valence remained positive and stable. CONCLUSIONS: On average, middle aged, formerly sedentary women selected an intensity that is considered physiologically effective and reported that it did not feel hard or unpleasant. Future research should examine the sources of interindividual variability and the consequences of exercising at an intensity that exceeds one's preferred level. PMID- 15530594 TI - Evaluation of a cognitive-behavioral, group, randomized controlled intervention trial to prevent sexually transmitted infections and unintended pregnancies in young women. AB - BACKGROUND: Few cognitive-behavioral interventions have focused on preventing sexually transmitted infections (STIs) and unintended pregnancies (UPs) in young, sexually active women in a single study. Military recruit training provides a well-defined, national, nonclinic sample in which to evaluate such an intervention. METHODS: All female Marine recruits (N=2,288) in training were approached. Of these, 2,157 (94.3%) voluntarily agreed and were randomly assigned, by platoons, to participate in cognitive-behavioral interventions to prevent STIs or UPs or to prevent physical training injuries and cancer. Participants completed self-administered questionnaires and were screened for pregnancy, Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis at baseline and, on average, 1 and 14 months postintervention. RESULTS: A higher proportion of the control group had a postintervention STI or UP [odds ratio (OR)=1.41, 95% confidence interval (CI)=1.01-1.98]. Among participants who had no history of STIs or pregnancy, but who engaged in risky sexual behaviors just before recruit training, the control group was more likely to acquire a postintervention STI (OR=3.24, CI=1.74-6.03). Among participants who were not sexually experienced at baseline, the control group was more likely to have casual (OR=2.05, 95% CI=1.04-4.08) and multiple (OR=1.87, 95% CI=1.01-3.47) sexual partners postintervention. CONCLUSIONS: This randomized controlled trial indicates that cognitive-behavioral interventions are effective for reducing behavioral risk and preventing STIs and UPs in young, sexually active women who are not seeking health care. PMID- 15530595 TI - The associations between leisure-time physical activity and inflammatory and coagulation markers related to cardiovascular disease: the ATTICA Study. AB - BACKGROUND: As coronary heart disease is increasingly seen as an inflammation process, we evaluated the hypothesis whether physical activity reduces coronary heart disease risk by modifying the levels of inflammatory and coagulation markers. METHODS: From May 2001 to December 2002, we randomly enrolled 1524 adult men and 1518 women, without any evidence of cardiovascular disease, stratified by age-gender (census 2001), from the greater area of Athens, Greece. We assessed the relationship between self-reported physical activity status and inflammation markers (high sensitivity C-reactive protein, serum amyloid-A, fibrinogen, interleukin-6, tumor necrosis factor-alpha, and white blood cell counts), after taking into account the effect of several confounders. RESULTS: Eight hundred seventy-four (57%) of men and 903 (59%) of women were classified as sedentary. Multivariate statistical analysis after adjustment for gender, age, smoking habits, body mass index, total cholesterol, blood glucose, and systolic and diastolic blood pressure levels showed that participants devoted to high physical activity (>7 kcal/min expended) had 29% lower levels of C-reactive protein, 19% of white blood cell counts, 22% lower concentrations of amyloid-A, 20% lower levels of tumor necrosis factor-alpha, 32% of interleukin-6, and 11% of fibrinogen (all P<0.05) as compared to those who were devoted to sedentary life. CONCLUSIONS: Our findings suggest that the adoption of a physically active lifestyle modifies the inflammation process in healthy individuals. PMID- 15530596 TI - Lifestyle and mental health. AB - STUDY OBJECTIVE: To examine the relationship between healthy lifestyles and self rated mental health. DESIGN: A cross-section telephone survey involving multiple callbacks. Using a Behavioral Risk Factor Surveillance Survey, information about lifestyle, demographic characteristics, and self-rated mental health was collected. SETTING: Amarillo, Texas, a city of approximately 170,000 in the Panhandle of Texas. PARTICIPANTS: Six hundred and twenty-one adults. MAIN RESULTS: Good mental health was independently associated with physical activity, being male, and being married. Poor mental health was associated with obesity, daily smoking, being Hispanic, and being non-Hispanic White. CONCLUSIONS: Programs targeting risk factors for poor physical health also may improve mental health in the community. PMID- 15530597 TI - A nationwide cohort study of educational background and major causes of death among the elderly population in Japan. AB - BACKGROUND: This prospective cohort study examined the association between educational level and major causes of death in Japan. METHOD: A baseline survey was conducted between 1988 and 1990 among 110,792 inhabitants of 45 areas aged 40 79 years. Follow-up surveys were conducted annually and causes of death were identified from death certificates. The analysis was restricted to 16,715 men and 23,284 women. RESULTS: During the follow-up period (377,139 person-years), 6628 deaths were recorded. Individuals with low levels of education had an increased overall risk of death [relative risk (RR)=1.16, 95% confidence interval (CI): 1.08, 1.25, in men; RR=1.26, 95% CI: 1.14, 1.39, in women], cancers (RR=1.17, 95% CI: 1.04, 1.32, in men; RR=1.10, 95% CI: 0.93, 1.30, in women), and death from external causes (RR=1.81, 95% CI: 1.29. 2.54, in men; RR=1.78, 95% CI: 1.18, 2.70, in women). Ischemic heart disease risk was marginally reduced in men with low levels of education (RR=0.77, 95% CI: 0.58, 1.01). CONCLUSIONS: These results show that health inequalities exist in Japan, even though wealth inequalities are relatively low. Social and political initiatives will be needed to correct these inequities between different socioeconomic statuses. PMID- 15530598 TI - What accounts for the association of education and smoking cessation? AB - BACKGROUND: Education has been identified as a potent sociodemographic predictor of smoking cessation and the Healthy People 2010 goals include the elimination of health disparities attributable to education. METHOD: The current study prospectively examined the association of education with smoking cessation as well variables that might account for that association among employed adults residing in the southeastern United States. RESULTS: A strong educational gradient in cessation was evident. Only 6% of smokers with less than a high school (HS) degree quit smoking during the 4-year study period, whereas 17% of smokers with a HS degree but no college degree and 28% of smokers with at least a college degree quit smoking. CONCLUSIONS: Education appeared to uniquely contribute to the prediction of smoking abstinence over and above the effects of demographic, environmental, tobacco dependence, transtheoretical model, and job related variables. Obtaining a better understanding of how or why education influences smoking cessation could contribute to reducing the educational gradient in abstinence and warrants further research attention. PMID- 15530599 TI - Factors associated with men's use of prostate-specific antigen screening: evidence from Health Information National Trends Survey. AB - BACKGROUND: Rapid uptake of prostate-specific antigen (PSA) testing has occurred in the United States despite inconclusive evidence regarding mortality benefit. METHODS: We examined data (n=927) from the 2003 Health Information National Trends Survey to assess prevalence of self-reported PSA use and its association with patients' decision making. RESULTS: Over half (55.2%) the sample reported ever having had a PSA test. Men aged 65-74 (OR=2.53, 1.49-4.31), with some college (OR=2.41, 1.22-4.77) or college degrees (OR=5.01, 2.53-9.90) were more likely to have had PSA tests, while men without health insurance (OR=0.32, 0.12 0.88) or a usual source of care (OR=0.35, 0.22-0.54) were less likely. In a model including healthcare provider communication and information seeking, men who reported that providers involved them in decisions (OR=1.76, 1.02-3.03) and recommended PSA (OR=236.3, 70.5-791.4) were more likely to have had the tests. Men aged 65-74 (OR=2.30, 1.33-4.00), with college degrees (OR=2.91, 1.45-5.82), and greater information attention/seeking (OR=1.23, 1.07-1.40) were more likely to report PSA recommendations, while those without usual care were less likely (OR=0.37, 0.22-0.64). Men without usual care (OR=0.38, 0.20-0.71) and Hispanic men (OR=0.40, 0.19-0.85) were less likely to report that healthcare providers involved them in healthcare decisions. CONCLUSIONS: Results emphasize the relevance of patient decision making and the importance of healthcare providers in PSA testing. PMID- 15530600 TI - Who are the elderly who never receive influenza immunization? AB - OBJECTIVE: This paper estimates the rates of lifetime nonreceipt of influenza immunization among elderly and examines variations in the lifetime nonreceipt of immunization by gender, race and ethnic group, socioeconomic status, access to health care, and health status. METHODS: Cross-sectional, nationally representative data on 5557 adults older than 50 years of age and living in the community from the 2000 Medical Expenditure Panel Survey are used. Lifetime nonreceipt of influenza immunization was analyzed with bivariate and multivariate statistical techniques. FINDINGS: Thirty-one percent of the elderly reported never receiving influenza immunization and 20% reported irregular immunization. Higher odds of lifetime nonreceipt of vaccination and irregular vaccination were seen among African-Americans, young-old, current smokers, and those with no usual source of care. CONCLUSIONS: Future campaigns to increase immunization rates should be tailored to target this hard-to-reach group of individuals. PMID- 15530601 TI - Does measurement dependence explain the effects of the Life Skills Training program on smoking outcomes? AB - BACKGROUND: The Life Skills Training (LST) program is the most prominent school based smoking prevention program in terms of its consistency in being named on lists of best practices. This study assessed whether the results pertaining to cigarette smoking reported in evaluations of the LST program are measurement dependent. METHODS: Seventeen reports published between 1980 and 2003 that included at least one outcome measure pertaining to cigarette smoking were identified. Data pertaining to the cigarette smoking measures used in the analysis and whether the results showed a statistically significant difference between experimental and control groups at follow-up were extracted from the reports. RESULTS: Fourteen different outcome measures were used across 17 reports. Only three pairs of reports presented the same set of outcomes. Recent reports showed the most consistent set of findings in support of the LST program, but there was little consistency in the outcome measures used in these analyses. CONCLUSIONS: The use of so many smoking outcomes in the LST program evaluations raises concern as to whether the positive program effects reported are measurement dependent. PMID- 15530602 TI - Public-health preparedness requires more than surveillance. PMID- 15530603 TI - Warning: geography may be hazardous to your health. PMID- 15530604 TI - Preventing disability in elderly people. PMID- 15530605 TI - Cervical screening in adolescents--at least do no harm. PMID- 15530606 TI - New hope for treatment of neonatal haemochromatosis. PMID- 15530607 TI - The rising tide of childhood type 1 diabetes--what is the elusive environmental trigger? PMID- 15530608 TI - Professionalism and the medical student. PMID- 15530610 TI - New therapeutic targets in multiple myeloma. PMID- 15530611 TI - Fighting for food aid--the struggle to assist groups affected by HIV/AIDS. PMID- 15530612 TI - The UK Draft Mental Health Bill 2004: a compromise that pleases no one. PMID- 15530614 TI - Historical keywords: trial. PMID- 15530615 TI - Role of aspirin in MATCH. PMID- 15530616 TI - Role of aspirin in MATCH. PMID- 15530618 TI - Role of aspirin in MATCH. PMID- 15530619 TI - Peripheral neuropathy. PMID- 15530620 TI - P450 oxidoreductase deficiency. PMID- 15530622 TI - Peripheral neuropathy. PMID- 15530624 TI - HIV transmission through health care in sub-Saharan Africa. PMID- 15530625 TI - Two good reasons to drop type 2 virus from oral polio vaccine. PMID- 15530627 TI - Population-based multidimensional assessment of older people in UK general practice: a cluster-randomised factorial trial. AB - BACKGROUND: The benefit of multidimensional assessment and management of older people remains controversial. Most trials have been too small to produce adequate evidence to inform policy. We aimed to measure the effects of different approaches to assessment and management of older people. METHODS: We undertook a cluster-randomised factorial trial in 106 general practices (43219 eligible patients aged 75 years and older, 78% participation), comparing (1) universal versus targeted assessment and (2) subsequent management by hospital outpatient geriatric team versus the primary-care team. All participants received a brief multidimensional assessment followed by a nurse-led in-depth assessment in the universal group, whereas in the targeted group the in-depth assessment was offered only to those with problems established at the brief assessment. Referrals to the randomised team (geriatric management or primary care), other medical or social services, health-care workers, or agencies, and emergency referrals to the general practitioner were based on a standard protocol at the in depth assessment. The primary endpoints were mortality, admissions to hospital and institution, and quality of life. Analysis was by intention to treat and per protocol. This trial has been assigned the International Standardised Randomised Controlled Trial Number ISRCTN23494848. FINDINGS: Mortality and hospital or institutional admissions did not differ between groups. During 3 years' follow up, significant improvements in quality of life resulted from universal versus targeted assessment in terms of homecare, and from management by geriatric team versus primary-care team, in terms of mobility, social interaction, and morale. However, only the result for social interaction was consistent with a small but important effect. INTERPRETATION: The different forms of multidimensional assessment offered almost no differences in patient outcome. PMID- 15530628 TI - Regression of low-grade squamous intra-epithelial lesions in young women. AB - BACKGROUND: The aim of this study was to assess the probability of low-grade squamous intra-epithelial lesion (LSIL) regression in young women, and to examine the factors associated with this regression. METHODS: In a longitudinal study of human papilloma virus (HPV) infection, female adolescents aged 13-22 years were examined every 4 months by cytology, colposcopy, and HPV DNA status. Both prevalent and incident LSIL cases were included in the analysis, with regression defined as at least three consecutive normal Pap smears. FINDINGS: Median follow up time from baseline (defined as the time of first LSIL diagnosis) for the 187 women with LSIL was 61 months (IQR 34-80). Median time they had been sexually active at diagnosis was 3.2 years (2.6-6.5). Probability of regression for the entire cohort was 61% (95% CI 53-70) at 12 months and 91% (84-99) at 36 months of follow-up. No associations were found between LSIL regression and HPV status at baseline, sexual behaviour, contraceptive use, substance or cigarette use, incident sexually transmitted infection, or biopsy. Multivariate analysis showed that only HPV status at the current visit was associated with rate of regression, whether infection was caused by one or more viral types (relative hazard=0.3 [95% CI 0.21-0.42], and 0.14 [0.08-0.25], respectively). INTERPRETATION: The high rate of regression recorded in this study lends support to observation by cytology in the management of LSIL in female adolescents. Negative HPV status was associated with regression, suggesting that HPV testing could be helpful in monitoring LSIL. PMID- 15530629 TI - Atenolol in hypertension: is it a wise choice? AB - BACKGROUND: Atenolol is one of the most widely used beta blockers clinically, and has often been used as a reference drug in randomised controlled trials of hypertension. However, questions have been raised about atenolol as the best reference drug for comparisons with other antihypertensives. Thus, our aim was to systematically review the effect of atenolol on cardiovascular morbidity and mortality in hypertensive patients. METHODS: Reports were identified through searches of The Cochrane Library, MEDLINE, relevant textbooks, and by personal communication with established researchers in hypertension. Randomised controlled trials that assessed the effect of atenolol on cardiovascular morbidity or mortality in patients with primary hypertension were included. FINDINGS: We identified four studies that compared atenolol with placebo or no treatment, and five that compared atenolol with other antihypertensive drugs. Despite major differences in blood pressure lowering, there were no outcome differences between atenolol and placebo in the four studies, comprising 6825 patients, who were followed up for a mean of 4.6 years on all-cause mortality (relative risk 1.01 [95% CI 0.89-1.15]), cardiovascular mortality (0.99 [0.83-1.18]), or myocardial infarction (0.99 [0.83-1.19]). The risk of stroke, however, tended to be lower in the atenolol than in the placebo group (0.85 [0.72-1.01]). When atenolol was compared with other antihypertensives, there were no major differences in blood pressure lowering between the treatment arms. Our meta-analysis showed a significantly higher mortality (1.13 [1.02-1.25]) with atenolol treatment than with other active treatment, in the five studies comprising 17671 patients who were followed up for a mean of 4.6 years. Moreover, cardiovascular mortality also tended to be higher with atenolol treatment than with other antihypertensive treatment. Stroke was also more frequent with atenolol treatment. INTERPRETATION: Our results cast doubts on atenolol as a suitable drug for hypertensive patients. Moreover, they challenge the use of atenolol as a reference drug in outcome trials in hypertension. PMID- 15530630 TI - High-dose immunoglobulin during pregnancy for recurrent neonatal haemochromatosis. AB - BACKGROUND: Neonatal haemochromatosis is a rare disease of gestation that results in severe fetal liver injury. We hypothesised an alloimmune aetiology for the disorder on the basis of its high recurrence rate in sibships. In this study, we assessed the effectiveness in preventing or changing the severity of recurrent neonatal haemochromatosis of administering during pregnancy high-dose intravenous immunoglobulin (IVIG) derived from pooled serum of multiple donors. METHODS: Women whose most recent pregnancy ended in documented neonatal haemochromatosis were treated with IVIG, 1 g/kg bodyweight, weekly from the 18th week until the end of gestation in their subsequent pregnancy. The outcomes of treated pregnancies were compared with those of randomly selected previous affected pregnancies for each woman, which were used as historical controls. FINDINGS: 15 women were treated through 16 pregnancies. All pregnancies progressed uneventfully and resulted in live babies with normal physical examinations and birthweights that were appropriate for gestational age. 12 babies had evidence of liver involvement with neonatal haemochromatosis: 11 had higher than normal concentrations of serum alpha-fetoprotein and ferritin or serum alpha-fetoprotein alone, including four with coagulopathy (international normalised ratio >1.5), and one had coagulopathy alone. All babies survived with medical or no treatment and were healthy at follow-up within the past 6 months. In analysis on a per mother basis comparing outcomes of treated gestations with those of randomly selected previous affected gestations, gestational IVIG therapy was associated with better infant survival (15 good outcomes vs two in previous pregnancies; p=0.0009). INTERPRETATION: Treatment with high-dose IVIG during gestation appears to have modified recurrent neonatal haemochromatosis so that it was not lethal to the fetus or neonate. These results further support an alloimmune mechanism for recurrent neonatal haemochromatosis. PMID- 15530631 TI - The rising incidence of childhood type 1 diabetes and reduced contribution of high-risk HLA haplotypes. AB - The incidence of childhood type 1 diabetes has risen over the past 50 years. We compared the frequency of HLA class II haplotypes in 194 patients diagnosed more than 50 years ago and 582 age-matched and sex-matched individuals diagnosed between 1985 and 2002. The proportion of high-risk susceptibility genotypes was increased in the earlier cohort (p=0.003), especially in those diagnosed at age 5 years or younger, which is consistent with the hypothesis that the rise of type 1 diabetes is due to a major environmental effect. PMID- 15530632 TI - Cardiac pacing: the state of the art. AB - Permanent cardiac pacing remains the only effective treatment for chronic, symptomatic bradycardia. In recent years, the role of implantable pacing devices has expanded substantially. At the beginning of the 21st century, exciting developments in technology seem to happen at an exponential rate. Major advances have extended the use of pacing beyond the arrhythmia horizon. Such developments include dual-chamber pacers, rate-response algorithms, improved functionality of implantable cardioverter defibrillators, combinations of sensors for optimum physiological response, and advances in lead placement and extraction. Cardiac pacing is poised to help millions of patients worldwide to live better electrically. We review pacing studies of sick-sinus syndrome, neurocardiogenic syncope, hypertrophic obstructive cardiomyopathy, and cardiac resynchronisation therapy, which are common or controversial indications for cardiac pacing. We also look at the benefits and complications of implantation in specific arrhythmias, suitability of different pacing modes, and the role of permanent pacing in the management of patients with heart failure. PMID- 15530633 TI - Antibiotics or surgery for vesicoureteric reflux in children. AB - CONTEXT: 1-2% of children have vesicoureteric reflux (VUR). VUR occurs in 25-40% of children with acute pyelonephritis. VUR can lead to renal scarring, hypertension, and end-stage renal disease. The best form of treatment for children with VUR is debated: no treatment, long-term antibiotic prophylaxis, surgery, or a combination of antibiotic prophylaxis and surgery. In children with recurrent urinary tract infections (UTIs) and progressive renal damage, despite antibiotic prophylaxis, surgical correction of VUR, especially high-grade VUR, is generally recommended. STARTING POINT: Danielle Wheeler and colleagues recently did a meta-analysis of ten randomised controlled trials (964 children) to evaluate whether any intervention for VUR is better than no treatment (Cochrane Database Syst Rev 2004; 3: CD001532). The main endpoints were incidence of UTIs, new or progressive renal damage, renal growth, hypertension, and glomerular filtration rate. They concluded that it is uncertain whether the identification of children with VUR is associated with clinically important benefit. The additional benefit of surgery over antibiotics is small. WHERE NEXT? New strategies for management will require a tailored diagnostic and therapeutic approach, including non-invasive or less invasive diagnostic procedures, and a less aggressive therapeutic approach. Whether the common practice of cystourethrography as a first-line investigation is warranted needs evaluation. The goal of paediatricians in the future, to prevent kidney damage, will probably be prevention of renal parenchymal injury and not necessarily the correction of ureterovesical junction anomalies. Because two main clinical pictures of VUR (diagnosed prenatally or postnatally with different age and sex distribution) can be identified, boys and girls will probably be managed differently. The factors responsible for congenital and acquired renal injury in children with VUR need to be studied. PMID- 15530634 TI - Independent clinical research in Europe. PMID- 15530635 TI - A lesson from the third year. PMID- 15530636 TI - A horse-riding accident and haemolytic anaemia. PMID- 15530637 TI - Neuroprotection by tetracyclines. AB - The neuroprotective properties of tetracyclines have been clearly established in rodent models of acute and chronic neurodegeneration during the past few years. Recent findings have provided novel insights into the molecular and cellular mechanisms of protection of neurons and oligodendrocytes by tetracyclines. These advances have prompted several clinical trials with minocycline, the most effective tetracycline, which are still in their early phases. Thus, tetracyclines hold great promise as therapeutic agents for the treatment of human neurodegenerative diseases. PMID- 15530638 TI - Bad news from the brain: descending 5-HT pathways that control spinal pain processing. AB - The identification of opioid systems led to much of the early work on pain pharmacology being based on understanding inhibitory mechanisms of analgesia. However, hyperalgesia and allodynia are common clinical symptoms and therefore hyperexcitability must be a major component of pain. Thus, the emphasis of current research into pain has shifted to understanding excitatory pathways that underlie neuronal sensitization and potentiation. Although much evidence supports the presence of descending inhibitory mechanisms of pain, reports of facilitatory pathways from the brainstem have been scarce. In this article, we review evidence for facilitatory 5-HT pathways that link spinal cord and brainstem areas involved in mood and emotions. Because pain encompasses affective aspects, we suggest that these 5-HT pathways and other circuits are important in determining the levels of pain, the outcome of drug treatments and provide a mechanism whereby emotions can alter pain perception. PMID- 15530639 TI - The histamine H3 receptor as a novel therapeutic target for cognitive and sleep disorders. AB - Histamine H3 receptor pharmacology, functions and biochemistry are far from being fully understood; however, progress is being made. Activation of this Gi/GO protein-coupled receptor affects cognition, the sleep-wake cycle, obesity and epilepsy, which are physiological and pathological conditions that are the main focus of research into the therapeutic potential of selective H3 receptor ligands. This heterogeneity of targets can be reconciled partially by the fact that the histamine system constitutes one of the most important brain-activating systems and that H3 receptors regulate the activity of histamine and other neurotransmitter systems. Furthermore, the H3 receptor shows functional constitutive activity, polymorphisms in humans and rodents with a differential distribution of splice variants in the CNS, and potential coupling to different intracellular signal transduction mechanisms. In light of the genetic, pharmacological and functional complexity of the H3 receptor, the importance of the histamine system as a therapeutic target to control the sleep-wake cycle and cognitive disorders will be discussed. PMID- 15530640 TI - Ca2+ channel beta-subunits: structural insights AID our understanding. AB - It has taken 17 years from the first identification of a voltage-gated Ca2+ channel (CaV) beta-subunit as a band on a gel following purification of skeletal muscle dihydropyridine (DHP) receptors in 1987 to the publication of key information on the structures of Ca2+ channel beta-subunits. Three recent X-ray crystallographic studies have now solved the structures of the core domains of three Ca2+ channel beta-subunits. In this article, the properties of these cytoplasmic auxiliary subunits will first be summarized. Then the CaVbeta structures and the information they provide regarding how these proteins interact with the CaValpha1 subunit will be discussed and the possible implications of these new data for G-protein modulation of Ca2+ channels will be examined. PMID- 15530641 TI - The TRPM ion channel subfamily: molecular, biophysical and functional features. AB - Significant progress in the molecular and functional characterization of a subfamily of genes that encode melastatin-related transient receptor potential (TRPM) cation channels has been made during the past few years. This subgroup of the TRP superfamily of ion channels contains eight mammalian members and has isoforms in most eukaryotic organisms. The individual members of the TRPM subfamily have specific expression patterns and ion selectivity, and their specific gating and regulatory mechanisms are tailored to integrate multiple signaling pathways. The diverse functional properties of these channels have a profound effect on the regulation of ion homoeostasis by mediating direct influx of Ca2+, controlling Mg2+ entry, and determining the potential of the cell membrane. TRPM channels are involved in several physiological and pathological conditions in electrically excitable and non-excitable cells, which make them exciting targets for drug discovery. PMID- 15530642 TI - Complexity of the bi-directional neuroimmune junction in the spleen. AB - The spleen is a crucial secondary lymphoid organ for circulating infectious agents that is densely innervated by sympathetic nerve fibres. Sympathetic nerve endings contact immune cells within the spleen, particularly in areas of T cells and macrophages (building the neuroimmune junction). Neurotransmitters are released into the vicinity of nerve terminals and bind to specific postsynaptic receptors on the surface of these cells. Local bi-directionality exists through cytokines and neurotransmitters from immune cells that modulate the release of sympathetic neurotransmitters from nerve terminals. This complex 'dialog' depends on microenvironmental factors such as infectious agents, and this 'conversation' is needed to balance the function of both the sympathetic nerve terminal and the immune system. Activation of the sympathetic nervous system and also the resting sympathetic nervous tone are important for controlling innate and adaptive immune responses. PMID- 15530643 TI - Paracrine role for periadventitial adipose tissue in the regulation of arterial tone. AB - Recent studies propose a paracrine role for periadventitial adipose tissue in the regulation of vascular tone. This regulation depends on the anatomical integrity of the periadventitial adipose tissue and involves adipocyte-derived relaxing factor (ADRF). Although the nature of ADRF is largely unknown, it is released by periadventitial adipocytes and induces vasorelaxation by opening K+ channels in the plasma membrane of smooth muscle cells. Alterations in the paracrine role of periadventitial adipose tissue might have a role in vascular dysfunction in hypertension and metabolic disease. Therefore, understanding alterations in ADRF release and the K+ channels involved will help further our understanding of the increased cardiovascular risk and development of chronic vascular disease in obesity. Furthermore, ADRF and perhaps its putative targets might represent exciting new targets for the development of drugs to treat cardiovascular disorders. PMID- 15530644 TI - The multispecific organic anion transporter family: properties and pharmacological significance. AB - Physiological and pharmacological studies indicate that the renal and hepatic organic anion transport systems are responsible for the elimination of numerous compounds, such as drugs, environmental substances and metabolites of both endogenous and exogenous origins. Recently, the molecular identity of the organic anion transport system, the OAT family, was revealed. To date, six OAT members have been identified and shown to have important roles not only in detoxification in the kidneys, liver and brain, but also in the reabsorption of essential compounds such as urate. The OAT family members are closely associated with the pharmacokinetics, drug-drug interactions and toxicity of anionic substances such as nephrotoxic drugs and uremic toxins. The molecular characterization of the OAT family encoded by SLC22A will be discussed. PMID- 15530645 TI - Techniques: GPCR assembly, pharmacology and screening by flow cytometry. AB - Flow cytometers are well known for their ability to analyze and sort cells at high rates based on physiological responses and expression of protein markers. The potential for flow cytometry in G-protein-coupled receptor (GPCR) research, however, is less well appreciated. Potential applications include: (i) the homogenous discrimination of free and bound ligands or proteins in both cellular and microsphere-based assays; and (ii) multiplexed ('suspension array') analysis of cell responses and protein-protein interactions. Innovative sample-handling systems also provide sub-second resolution of interaction kinetics and 1 second per well throughput of microliter-sized samples from multiwell plates. Flow cytometric methods using microspheres for analysis of GPCRs that interact with intracellular and extracellular binding partners such as ligands, G proteins and kinases have been established. These analyses can produce quantitative pharmacological data analogous to radioligand assays, and, in some cases, the probes can be integrated into the assembly as fluorescent fusion proteins. PMID- 15530646 TI - Sex steroids reduce DNaseI accessibility of androgen receptor promoter in adult male mice brain. AB - We have previously reported that androgen receptor (AR) expression is inversely correlated to its promoter methylation and is regulated by sex steroids. As chromatin structure plays an important role in transcriptional regulation, the effect of sex steroids on DNaseI accessibility of chromatin of AR promoter was examined in the brain cortex of adult and old mice of both sexes. Nuclei were digested with different concentrations of DNaseI and the extracted DNA was further cleaved by PstI and analyzed by Southern hybridization with DIG-labeled 695-bp AR promoter. With 50 U DNaseI, the intensity of PstI-specific 1.45-kb band was lower in intact female as compared to male groups, suggesting increased nuclease accessibility in female than male. Although gonadectomy increased DNaseI accessibility remarkably in male and female of both ages, testosterone decreased the accessibility in adult but increased in old male. Estradiol, on the other hand, decreased DNaseI accessibility in both adult male and old female but increased in old male and adult female. Thus, these findings suggest that the chromatin conformation of AR promoter varies with age and sex and its accessibility to DNaseI is reduced by testosterone and estradiol in the brain cortex of adult male mice. PMID- 15530647 TI - Nuclear factor-kappa B p65 in NMDA-induced retinal neurotoxicity. AB - Transcription factors of the nuclear factor-kappa B (NF-kappaB)/Rel family may be involved in neuronal cell death or survival. We examined the role of NF-kappaB p65 in N-methyl-D-aspartate (NMDA)-induced neurotoxicity in the rat retina. Western blot analysis showed that elevated levels of retinal NF-kappaB p65 protein at days 1 and 5 after intravitreal NMDA injection. Immunohistochemistry localized increased NF-kappaB p65 immunoreactivity in the ganglion cell layer (GCL) and the inner nuclear layer (INL) after NMDA injection especially in retinal ganglion cells (RGCs), displaced amacrine cells, and amacrine cells. Concomitant with the early increase in NF-kappaB p65 protein levels, there was an increase in NF-kappaB DNA binding activity after NMDA injection as shown by electrophoretic mobility shift assay (EMSA). These increases in NF-kappaB p65 protein levels and NF-kappaB DNA binding activity were totally abolished by simultaneous injection of NF-kappaB p65 antisense oligodeoxynucleotide (AS ODN). A partial but significant protective effect on the inner retina was noted when the AS ODN was given together with NMDA as shown by morphological analysis, morphometry of cells in the GCL and morphometry of inner plexiform layer thickness as well as quantitative real-time PCR of Thy-1 mRNA levels. These results suggest that activated NF-kappaB p65 may participate in NMDA-induced retinal neuronal cell death and that inhibition of NF-kappaB activation such as the use of AS ODN may be a viable neuroprotective strategy for protective RGCs and other inner retinal neurons. PMID- 15530648 TI - The effects of acute stress on the regulation of central and basolateral amygdala CRF-binding protein gene expression. AB - Corticotropin-releasing factor (CRF) is a key mediator of the behavioral, autonomic, and endocrine responses to stress. CRF binds two receptors and a CRF binding protein (CRF-BP), which may inactivate or modulate the actions of CRF at its receptors. The amygdala is an important anatomical substrate for CRF and contains CRF, its receptors, and CRF-BP. Few studies have examined the effects of acute stress on the regulation of amygdala CRF-BP with other CRF system genes. Therefore, we examined the time course of the effects of acute restraint stress on central (CeA) and basolateral (BLA) amygdala CRF system genes. Consistent with our previous study, acute stress increased BLA CRF-BP mRNA shortly after stress offset. Surprisingly, BLA CRF-BP mRNA remained elevated up to 21 h after the stressor. This effect was selective in the BLA as stress did not alter CeA CRF-BP mRNA, and there were no changes in CRF or CRF receptor mRNAs in either amygdala nucleus. These results suggest that alterations in BLA CRF-BP gene expression are a primary response of the BLA/CeA CRF system to acute stress. Because CRF-BP can modulate CRF action, changes in amygdala CRF-BP levels after stress exposure may affect the ability of an organism to adapt to future stressors. PMID- 15530649 TI - Plasma membrane calcium ATPase expression in the rat spinal cord. AB - Plasma membrane Ca2+-ATPase (PMCA) is a calcium pump that exists on the plasma membrane and has a role in keeping the intracellular Ca2+ concentration low. In the current study, the expression of PMCA isoforms in spinal cord tissues was investigated in detail and the changes of the expression was examined after contusion injury. Rats received a weight drop on the thoracic spinal cord as the injury or they received a sham surgery as a control. Three or twenty-four hours after spinal cord injury (SCI), the spinal cord was removed and processed for in situ hybridization, reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. PMCA1-4 mRNAs were expressed in neurons in the control spinal cord. Each isoform of the PMCA proteins showed distinct expression patterns in the spinal cord. PMCA1 and PMCA3 were expressed in all of the layers of gray matter. PMCA2 was also abundant in gray matter, except laminae I and II, while PMCA4 expression was restricted to the superficial layers of the dorsal horn. Distinct expression patterns of the PMCA isoforms suggest differential functions of each isoform in the spinal cord. After spinal cord injury, the expression of PMCA2 was decreased; however, the change in expression of other isoforms showed a tendency of decrease but did not reach a statistically significant level. The decrease in PMCA expression may contribute to the increase in intracellular Ca2+ concentration and PMCA may have a role in secondary injury following spinal cord injury. PMID- 15530650 TI - Neuroprotective gene expression profiles in ischemic cortical cultures preconditioned with IGF-1 or bFGF. AB - The mechanisms underlying growth factor preconditioning of neurons are only partially elucidated, and no studies have been conducted in this area using a gene profiling approach. We used cDNA microarrays to compare the transcriptional profiles of cells preconditioned either with insulin-like growth factor I (IGF-1) or basic fibroblast growth factor (bFGF), to identify differentially regulated genes that may function in growth factor signaling, response to oxygen-glucose deprivation (OGD), and most importantly, cell survival. Primary rat cortical cultures were treated with bFGF or IGF-1 for 2, 24, or 24 h followed by OGD for 90 min, and compared with cells that were subject to OGD without growth factor pretreatment. Although the majority of surveyed genes were unchanged in all experimental treatments, 175 genes (10% of the cDNAs on the chip) were found to be differentially regulated in at least one of the treatment conditions. Hierarchical clustering of these 175 genes was used to identify four expression clusters: IGF-1 regulated, bFGF regulated, OGD regulated, and putative neuroprotective genes. Further analysis using realtime RT-PCR confirmed that we had identified genes that are regulated by single growth factors, as well as several more that are co-regulated by both IGF-1 and bFGF. These genes can influence neuronal survival by affecting diverse pathways such as growth factor signal transduction (CD44, DTR, DUSP6, EPS8, IGFBP3), DNA repair and transcription (FOXJ1), metabolic homeostasis (RASA1, SHMT2), cytoskeletal stability (MSN, MAPT) and cholesterol biosynthesis (FDFT1, FDPS). PMID- 15530651 TI - Identification of putative rat ribonuclease III by differential display: a novel rat mRNA expressed in a circadian manner in the rat suprachiasmatic nucleus. AB - The suprachiasmatic nucleus (SCN) of the hypothalamus constitutes the principal site responsible for the generation and entrainment of circadian rhythms in mammals. The mechanisms of the circadian clock involve periodic gene expression. Here we report the use of differential display reverse transcriptase polymerase chain reaction to identify a novel rat mRNA sequence which is highly homologous to human ribonuclease III. Analysis of its expression in the rat brain by in situ hybridization histochemistry showed this transcript to be expressed at differing intensities at various sites. Temporal variation in expression was observed in the SCN, with a peak at circadian time (CT) 2 and a nadir at CT14. No significant changes in its expression were detected across the cycle within the supraoptic nucleus, cingulate cortex or caudate putamen. PMID- 15530652 TI - Hippocampal synaptic plasticity in mice devoid of cellular prion protein. AB - The cellular prion protein plays a role in the etiology of transmissible and inherited spongiform encephalopathies. However, the physiological role of the cellular prion protein is still under debate. Results regarding the synaptic transmission using the same strain of animals where the cellular prion protein gene was ablated are controversial, and need further investigation. In this work, we have studied the hippocampal synaptic transmission in mice devoid of normal cellular prion protein, and have shown that these animals present an increased excitability in this area by the lower threshold (20 Hz) to generate long-term potentiation (LTP) in hippocampal dentate gyrus when compared to wild-type animals. The mice devoid of normal cellular prion protein are also more sensitive to the blocking effects of dizocilpine and 2-amino-5-phosphonopentanoic acid on the hippocampal long-term potentiation generation. In situ hydridization experiments demonstrated overexpression of the mRNAs for the N-methyl-D-aspartate (NMDA) receptor NR2A and NR2B subunits in mice devoid of normal cellular prion protein. Therefore, our results indicate that these animals have an increased hippocampal synaptic plasticity which can be explained by a facilitated glutamatergic transmission. The higher expression of specific N-methyl-d aspartate receptor subunits may account for these effects. PMID- 15530654 TI - Induction of granulocyte colony-stimulating factor mRNA by focal cerebral ischemia and cortical spreading depression. AB - Granulocyte colony-stimulating factor (G-CSF) is a hematopoietic growth factor with neuroprotective and antiinflammatory properties. By real-time polymerase chain reaction we show that G-CSF transcripts are induced 485-fold at 4 h and 65 fold at 16 h in ischemic lesions after middle cerebral artery occlusion compared to control brains. Further analysis in photochemically induced focal ischemia revealed that G-CSF induction involved both the infarct area and remote nonischemic brain regions. Remote responses could be blocked by the noncompetitive NMDA receptor antagonist MK-801, suggesting periinfarct depolarizations as a trigger. To further confirm this notion, cortical spreading depression (CSD) was induced by focal application of KCl to the brain surface. CSD led to a 90-fold increase in G-CSF mRNA. Contrastingly, the induction of granulocyte-monocyte (GM)-CSF, another member of the hematopoietic growth factor family, was only moderate (sixfold) and restricted to ischemic brain lesions. In conclusion, G-CSF induction in the brain may be part of an intrinsic stress response aimed at limitation of neuronal damage. PMID- 15530653 TI - L-type Ca2+ channel blockers promote Ca2+ accumulation when dopamine receptors are activated in striatal neurons. AB - Dopamine (DA) receptor-mediated signal transduction and gene expression play a central role in many brain disorders from schizophrenia to Parkinson's disease to addiction. While trying to evaluate the role of L-type Ca2+ channels in dopamine D1 receptor-mediated phosphorylation of the transcription factor cyclic AMP response element-binding protein (CREB), we found that activation of dopamine D1 receptors alters the properties of L-type Ca2+ channel inhibitors and turns them into facilitators of Ca2+ influx. In D1 receptor-stimulated neurons, L-type Ca2+ channel blockers promote cytosolic Ca2+ accumulation. This leads to the activation of a molecular signal transduction pathway and CREB phosphorylation. In the absence of dopamine receptor stimulation, L-type Ca2+ channel blockers inhibit CREB phosphorylation. The effect of dopamine on L-type Ca2+ channel blockers is dependent on protein kinase A (PKA), suggesting that protein phosphorylation plays a role in this phenomenon. Because of the adverse effect of activated dopamine receptors on L-type Ca2+ channel blocker action, the role of L type Ca2+ channels in the dopamine D1 receptor signal transduction pathway cannot be assessed with pharmacological tools. However, with antisense technology, we demonstrate that L-type Ca2+ channels contribute to D1 receptor-mediated CREB phosphorylation. We conclude that the D1 receptor signal transduction pathway depends on L-type Ca2+ channels to mediate CREB phosphorylation. PMID- 15530656 TI - Allele-specific overexpression in astrocytes of an Alzheimer's disease associated alpha-1-antichymotrypsin promoter polymorphism. AB - Alpha-1-antichymotrypsin (ACT), a serine proteinase inhibitor is synthesised predominantly in the liver and in other tissues including the brain. ACT is a major component of the senile plaques (SP) characteristic of Alzheimer's disease (AD). Increased production locally in the brain may be associated with Alzheimer's disease as ACT acts as a "pathological chaperone", promoting beta amyloid assembly into neurotoxic fibrils. Recent reports suggest that the T allele of a G/T polymorphism at position -51 of ACT is associated with cognitive decline in AD patients. We demonstrate that the T allele is markedly overexpressed by almost 225% in an astrocytic cell line in response to oncostatin M (OSM) compared with a 35% increase in a mixed population of brain-derived cells. This effect is probably mediated by a higher binding affinity of the transcription initiation complex to the higher expressing allele and overexpression of ACT by astrocytes could thus contribute to increased beta amyloid fibril formation in AD. PMID- 15530655 TI - Photoperiodic regulation of androgen receptor and steroid receptor coactivator-1 in Siberian hamster brain. AB - Seasonal changes in the neuroendocrine actions of gonadal steroid hormones are triggered by fluctuations in daylength. The mechanisms responsible for photoperiodic influences upon the feedback and behavioral effects of testosterone in Siberian hamsters are poorly understood. We hypothesized that daylength regulates the expression of androgen receptor (AR) and/or steroid receptor coactivator-1 (SRC-1) in specific forebrain regions. Hamsters were castrated and implanted with either oil-filled capsules or low doses of testosterone; half of the animals remained in 16L/8D and the rest were kept in 10L/14D for the ensuing 70 days. The number of AR-immunoreactive (AR-ir) cells was regulated by testosterone in medial amygdala and caudal arcuate, and by photoperiod in the medial preoptic nucleus and the posterodorsal medial amygdala. A significant interaction between photoperiod and androgen treatment was found in medial preoptic nucleus and posterodorsal medial amygdala. The molecular weight and distribution of SRC-1 were similar to reports in other rodent species, and short days reduced the number of SRC-1-ir cells in posteromedial bed nucleus of the stria terminalis (BNST) and posterodorsal medial amygdala. A significant interaction between androgen treatment and daylength in regulation of SRC-1-ir was found in anterior medial amygdala. The present results indicate that daylength-induced fluctuations in SRC-1 and AR expression may contribute to seasonally changing effects of testosterone. PMID- 15530657 TI - Regulatory role of dADAR in ROS metabolism in Drosophila CNS. AB - Pre-mRNA adenosine deaminase (ADAR) is involved in many physiological processes by either directly converting adenosine to inosine in certain pre-mRNAs or indirectly regulating expression of certain genes. Mutations of Drosophila ADAR (dADAR) results in neuronal dysfunction and hypersensitivity to oxygen deprivation. Recently, we found that the mutant flies were very resistant to paraquat, a compound that generates free radicals. In order to further characterize the neuronal role of dADAR and understand the basis for the resistance to the oxidative stress, we investigated the effect of dADAR on the expression of genes encoding scavengers of cellular reactive oxygen species (ROS) in both dADAR mutant and overexpression flies. Our data show that the expression of the genes encoding known ROS scavengers [superoxide dismutase (SOD) and catalase] is not regulated by dADAR. However, the transcripts of genes encoding two potential ROS scavengers (dhd and Cyp4g1) were robustly increased in dADAR mutant flies, and conversely both were significantly decreased in dADAR overexpressing flies. Using dhd [encoding a Drosophila homolog of the mammalian protein thioredoxin (Trx)] transgenic flies, we confirmed that the resistance of dADAR mutant flies to paraquat resulted, at least partially, from the up regulation of dhd gene in dADAR mutant flies. Our data not only confirm the importance of ADAR in maintenance of neuronal function but also reveal its regulatory role in the expression of genes encoding ROS scavengers. PMID- 15530658 TI - Stress induced changes in cortical and hypothalamic c-fos expression are altered in fragile X mutant mice. AB - Fragile X (FraX) syndrome is characterized by mental retardation and a behavioral phenotype that includes stress-related behaviors. Recently, FraX children were shown to have elevated glucocorticoid hormones under basal conditions and an exaggerated hormonal response to stress. In the present study, fragile X mental retardation 1-knockout (Fmr1-KO) and wild-type (WT) mice were subjected to immobilization stress for 30 min or 2 h, killed with paired controls, and the hippocampus, neocortex, and hypothalamic paraventricular nucleus (PVN) assessed by in situ hybridization for effects on c-fos mRNA. The main effect of stress in hippocampus was a reduction in mRNA levels within CA3-CA1 pyramidal cells in both genotypes. Stress significantly reduced CA1 c-fos mRNA in Fmr1-KOs at 30 min ( 41%) and 2 h (-57%), whereas in WTs levels were significantly reduced only at 2 h (-57%). In neocortex, 30 min stress significantly increased c-fos mRNA in Fmr1 KOs only (+53%); however, by 2 h levels were reduced in both genotypes versus respective controls. In the paraventricular nucleus, c-fos mRNA levels were significantly, and equally, increased in both genotypes at 30 min. However, at 2 h, mRNA levels were still elevated in the Fmr1-KOs, whereas they had returned to control values in the WTs. Finally, immobilization stress significantly increased serum corticosterone levels in both genotypes at 30 min and 2 h, with Fmr1-KOs exhibiting greater levels than WTs; levels were statistically different at 2 h. These data indicate a greater response to stress in FraX mutants than in WTs, and further support the hypothesis of a dysregulated hypothalamic-pituitary-adrenal (HPA) axis in FraX syndrome. PMID- 15530659 TI - Repeated 4-aminopyridine seizures reduce parvalbumin content in the medial mammillary nucleus of the rat brain. AB - Parvalbumin (Pv) containing fast spiking neurons play a crucial role in synchronizing the activity of excitatory neuronal circuits in the brain. Alterations of parvalbumin content in these neurons can affect their spike characteristics and, ultimately, may increase the susceptibility of neuronal circuits to epileptic seizures. In the present study, we examined whether repeated 4-aminopyridine (4-AP)-induced seizures modify the regional parvalbumin contents in the rat brain. 4-Aminopyridine was injected intraperitoneally in adult rats, controls received the solvent. Animals were sacrificed at 3 h after a single acute treatment, or following repeated, daily treatments of 12 days. In situ hybridization (ISH) indicated significantly decreased parvalbumin mRNA level in the medial mammillary nucleus (MM) at 12 days. Western blotting revealed 20.1% significant decrease of parvalbumin content in the medial mammillary area, while parvalbumin immunohistochemistry indicated no change of the number of immunoreactive cells in the medial mammillary nucleus. The results reveal the downregulation of the transcription of the parvalbumin gene and the decrease of parvalbumin synthesis in medial mammillary nucleus neurons in response to experimental seizures. PMID- 15530660 TI - Differential regulation of the regulatory subunits for phosphatidylinositol 3 kinase in response to motor nerve injury. AB - Type Ia phosphatidylinositol 3-kinase (PI3K) generates lipid products that operate as one of major second messengers following activation of tyrosine kinase receptors. PI3K is a heterodimer composed of a 110-kDa catalytic subunit and a regulatory subunit. In this study, we determined the expression of mRNA for the regulatory subunits after injury of rat hypoglossal nerves. In situ hybridization histochemistry revealed that the expression of PI3K regulatory subunit alpha isoforms (p85alpha, p55alpha, and p50alpha) was significantly enhanced in injured motor neurons, whereas other regulatory subunits such as p85beta or p55gamma were not detected. Of the alpha isoforms, the greatest increase was observed in p55alpha mRNA levels, while there were smaller increases in p85alpha and p50alpha mRNA expression. These results were confirmed by RT-PCR analysis. Further immunohistochemical analysis also confirmed the increased level of p55alpha protein in injured motor neurons. Taken together with the previously reported induction of the p110alpha catalytic subunit in injured neurons, these results suggest that PI3K, consisting of p55alpha and p110alpha, plays a crucial role in the process of nerve regeneration. PMID- 15530661 TI - AF1q, a differentially expressed gene during neuronal differentiation, transforms HEK cells into neuron-like cells. AB - Three cell groups, neural stem/progenitor cells (NS/PCs) dissociated from the embryonic day 11 (E11) rodent cerebral cortex, expanded NS/PC cultures, and cultured neurons from E15, were used to conduct a genomic study with differential display (DD). The mouse Af1q, homologue of human AF1q, was found to be significantly up-regulated during the neuronal production from NS/PCs. The ectopic expression of human AF1q triggered the expression of the neuronal marker TuJ1 in non-neuronal human embryonic kidney (HEK) cells. PMID- 15530662 TI - Upregulation of amyloid precursor protein isoforms containing Kunitz protease inhibitor in dementia with Lewy bodies. AB - Amyloid precursor protein (APP) is involved in the accumulation of alpha synuclein, the main component of Lewy bodies. It is currently unknown, however, whether any of the APP isoforms is instrumental in alpha-synuclein deposition in dementia with Lewy bodies (DLB). Using real-time RT-PCR, we have studied relative mRNA expression levels of APP isoforms in frozen postmortem frontal cortices of DLB patients, Alzheimer disease (AD) patients, and control subjects. Of the three main APP isoforms, the two with a Kunitz protease inhibitory (KPI) motif (APP770 and APP751) were found to be specifically overexpressed in the frontal cortices of DLB patients when compared with controls and AD patients. These findings suggest a specific role of APP isoforms containing Kunitz protease inhibitor in DLB pathogenesis. PMID- 15530663 TI - Galantamine blocks cloned Kv2.1, but not Kv1.5 potassium channels. AB - Galantamine is a cholinesterase inhibitor (AChEI) currently used in treatment of Alzheimer's disease (AD). In the present study, the effects of galantamine on currents of cloned Kv2.1 and Kv1.5 potassium channels were investigated by using patch-clamp whole cell recording techniques. Kv2.1 and Kv1.5 were stably expressed in HEK293 cells. Galantamine blocked Kv2.1 current in a concentration dependent manner. When depolarizing from -50 to +40 mV, the IC50 of galantamine for inhibition of Kv2.1 was 5.6 microM. Galantamine 10 microM shifted the activation curve of Kv2.1 to negative potential by 4.0 mV. At the same concentration, galantamine shifted the inactivation curve to negative potential by 25.2 mV. While Kv1.5 was not sensitive to galantamine, Kv1.5 current was not changed by galantamine at concentration of 10 microM. Our data suggest that galantamine potently blocks Kv2.1, but not Kv1.5 channels. PMID- 15530664 TI - MK-801 upregulates the expression of d-amino acid oxidase mRNA in rat brain. AB - We have evaluated the effect of the systemic administration of MK-801 (0.4 mg/kg) on the gene expression of D-amino acid oxidase (DAO) in several brain areas of the rat. The levels of DAO mRNA in all the brain areas significantly increased and peaked at 4 h after the administration. The present results suggest that there is a link between the expression of DAO mRNA and the N-methyl-D-aspartate (NMDA) receptor activity. PMID- 15530665 TI - Continuous ethanol exposure inhibits agonist-stimulated phosphorylation of p70S6 kinase and ribosomal S6 protein in cultured rat astrocytes. AB - The effects of ethanol exposure on agonist-stimulated phosphorylation of p70S6 kinase and ribosomal S6 protein were determined in confluent astrocyte monolayers. Basal phosphorylation of p70S6 kinase and S6 protein was either unaffected or reduced, respectively, after exposure to 50 mM ethanol for 4 days. The abilities of norepinephrine, carbachol and epidermal growth factor to phosphorylate these proteins were significantly decreased after ethanol exposure. In contrast, ethanol exposure had no effect on the protein expression of either p70S6 kinase or S6 protein. Our data suggest that continuous ethanol exposure results in a generalized decrease in agonist-activation of the p70S6 pathway. PMID- 15530666 TI - Letter to the editor. The delivery of influenza vaccinations and the role of pharmacists. PMID- 15530667 TI - Compassion for the animals, no concern for the children. PMID- 15530668 TI - Letter to the editor. The safety of thimerosal in newborn and infant vaccines. PMID- 15530670 TI - Sero-epidemiology of measles and mumps in Korea: impact of the catch-up campaign on measles immunity. AB - A catch-up campaign targeting children aged 8-16 years using measles-rubella (MR) vaccine was conducted during 2001 in Korea. To evaluate the impact of the campaign and assess mumps immunity, human IgG antibodies were detected using ELISA for measles (5826 samples) and mumps (5890 samples) in a national sample of opportunistically collected sera from a population aged 0-34 years. The measles immunity increased by 5-10% following the catch-up campaign in the targeted age group. Infants lost maternal antibodies rapidly and about 90% of infants were susceptible to measles and mumps at 6-8 months of life. The sero-prevalence of mumps antibody increased slowly with age and stabilized at a lower level when compared with that of measles. Despite an immediate reduction in susceptibility among the targeted age group of the catch-up campaign, continuous efforts to increase immunization coverage are needed to interrupt indigenous measles transmission. Furthermore, our results suggest continuous mumps outbreaks could occur because of the accumulation of susceptible individuals. PMID- 15530669 TI - Effectiveness of the MF59-adjuvanted influenza vaccine in preventing emergency admissions for pneumonia in the elderly over 64 years of age. AB - Case-control study designed to determine the effectiveness of an MF59-adjuvanted influenza vaccine in the population aged 65 years and older living in the community. Detailed health histories were obtained on both cases and controls that included a functional measure of co-morbidity (Barthel Index). Subjects were all eligible persons admitted to various hospitals with a diagnosis of pneumonia during the winter months and were matched by sex, hospital and admission week to controls admitted for non-medical reasons. The influenza vaccination programme using the MF59-adjuvanted influenza vaccine significantly reduced the probability of being hospitalised for pneumonia in the elderly over 64 years of age, even in a season with a low influenza activity, during which the predominant circulating strains were types B and A (H1N1). PMID- 15530671 TI - Salivary antibodies induced by the seven-valent PncCRM conjugate vaccine in the Finnish Otitis Media Vaccine Trial. AB - We studied salivary antibodies induced by a seven-valent pneumococcal conjugate vaccine (PncCRM). Healthy Finnish children (n=115), a subcohort of the Finnish Otitis Media (FinOM) Vaccine Trial, were immunised either with the PncCRM or a control vaccine (hepatitis B) at the age of 2, 4, 6, and 12 months. Salivary IgG, IgA, IgA1, IgA2 and sIg for serotypes 6B, 14, 19F, and 23F were measured at 7 and 13 months of age, and IgG and IgA also at 4-5 years of age. The PncCRM could induce both salivary anti-Pnc polysaccharide IgG and IgA. However, by the age of 4-5 years IgA concentrations had increased in both groups and were similar. The increases in IgA concentrations were mostly of IgA1 subclass. The difference between the PncCRM and the control group was more notable for serotypes 6B, 14 and 23F than for serotype 19F. We could not find evidence for the development of mucosal immunologic memory after vaccination with the PncCRM. PMID- 15530672 TI - Chemically synthesized protein as tumour-specific vaccine: immunogenicity and efficacy of synthetic HPV16 E7 in the TC-1 mouse tumour model. AB - Many successful candidate vaccines capable of combating tumours in animal models come to an untimely end because of the costs associated with the approval and production of the GMP-grade materials, which are usually of biological origin, for use in humans. We have used a GMP-compatible method to chemically synthesize a pure synthetic E7 protein of the human papillomavirus type 16 (HPV16-E7). This oncogen-derived protein is constitutively expressed in cervical cancer and its precursors and is thus considered as an excellent target for tumour-specific immunity. Injection of a mixture of the synthetic HPV16-E7 protein and the synthetic adjuvant CpG in mice resulted in strong functional HPV16-specific cytotoxic T-lymphocyte responses as measured by CD8+ MHC class I-tetramer staining, the detection of antigen-specific intracellular IFNgamma production and the ability to protect mice against a challenge with HPV16-E7+ TC-1 tumour cells in both prophylactic and therapeutic vaccination regimens. Our results demonstrate the potential use of pure synthetic vaccines that can be efficiently produced under GMP at low cost, which will stimulate the translation of new vaccination strategies into phase I/II clinical trials. PMID- 15530673 TI - TBE booster immunization according to the rapid immunization schedule: are 3-year booster intervals really necessary? AB - In order to evaluate the need for further booster immunizations, 222 subjects aged 20-52 years, who had received the first booster dose with a new tick-borne encephalitis (TBE) vaccine in a preceding study, were invited for a serological follow-up. A total of 191 and 182 adult subjects were analyzed for the persistence of neutralizing TBE antibodies at 1 and 2 years following the first booster immunization, respectively. Both serological follow-ups revealed high levels of neutralizing TBE antibodies in more than 99% of subjects. Although an expected decline of the respective geometric mean titers (GMTs) was noted after booster immunization, the titers were still far above the values noted after primary immunization at the 2-year follow-up. The kinetic curve clearly indicates a longer persistence of neutralizing TBE antibodies than currently expected. To conclude, these results suggest that the administration of a further booster dose 3 years after the first one (according to current recommendations) does not seem to be necessary in this study population. PMID- 15530674 TI - Safety and immunogenicity profile of an experimental hepatitis B vaccine adjuvanted with AS04. AB - The reactogenicity and safety of an experimental hepatitis B (HB) vaccine containing adjuvant system (AS04) was compared with a licensed vaccine in a phase III, single-blind, randomised study in healthy volunteers >or=15 years of age. A total of 1303 subjects were enrolled to receive either two doses of HB-AS04 (0, 6 months) or three doses of the comparator vaccine (0, 1, 6 months). Two doses of HB-AS04 elicited seroprotection rates close to 100% and two-fold higher GMTs than the comparator vaccine. Results showed that both vaccines were well tolerated and the general safety profile of HB-AS04 was similar to that of the comparator vaccine. PMID- 15530675 TI - Determination of immunity to vaccinia virus by using diluted inactivated vaccinia vaccine solution for skin testing. AB - AIMS: We evaluated the usefulness of skin test prepared by inactivation of vaccinia vaccine in predicting immunity to vaccinia. Skin test was injected to 77 healthy adults. Twenty had a recent smallpox vaccination (group 1). Thirty-seven were long term vacinees (group 2), while 20 subjects had never been vaccinated for smallpox (group 3). RESULTS: Mean size of induration was 7.9, 5.3 and 0.4 mm for subjects from groups 1, 2 and 3, respectively (P<0.03 for difference between groups). Induration >or=5 mm correlated with neutralizing antibody titer >1:32 (r=0.73, P<0.0001). CONCLUSIONS: The skin test is a potentially useful tool for the assessment of immunity to vaccinia. PMID- 15530676 TI - Efficacy of postexposure immunization with live attenuated varicella vaccine in the household setting--a pilot study. AB - The aim of the present study was to examine the efficacy of postexposure vaccination with Varilrix in the household setting. A randomized, double-blind, placebo-controlled design was used. Twenty-two children received the varicella vaccine and 20, a placebo. The relative risk of developing varicella with a placebo compared with the vaccine was 1.1 (95% confidence interval 0.55-2.21). The risk of developing moderate to severe disease was eight times greater in the placebo group (RR=8), indicating an 80% protective effect against moderate/severe disease. The varicella vaccine Varilrix may not be effective in preventing varicella when administered after household exposure, although it is highly effective in ameliorating the disease in those who acquire it under these circumstances. PMID- 15530677 TI - Influence of complement C3 amount on IgG responses in early life: immunization with C3b-conjugated antigen increases murine neonatal antibody responses. AB - Complement component C3, which plays an important role in both the innate and adaptative immune response, is present at low level in human infants. We show here that: (i) serum C3 amount is weak also in infant mice, (ii) these young animals fail to upregulate C3 to adult levels following tetanus toxoid immunization, (iii) neonatal macrophages have a limited capacity to synthesize C3 upon LPS exposure, (iv) conjugation of antigen to C3b significantly enhances antibody response elicited in 1-week-old mice--although it does not increase primary IgG response in adult mice. Altogether, this identifies C3 as one of the factors limiting early life antibody response and emphasizes the potential interest of immunization strategies overcoming this limitation. PMID- 15530678 TI - Anti-diphtheria antibody seroprotection rates are similar 10 years after vaccination with dTpa or DTPa using a mathematical model. AB - The reduced antigen content diphtheria, tetanus and pertussis (dTpa) vaccine (Boostrixtrade mark) has been shown to induce a strong booster response to all the vaccine components in 4-6 year olds. However, anti-diphtheria antibody levels were observed to be lower when compared to the "full strength" paediatric DTPa vaccine. To assess the impact of this difference on long-term protection, a mathematical model was developed to predict diphtheria antibody decay over time. The model was based on a linear decrease in log-transformed antibody concentrations after the first year post-vaccination. When applied to data collected 3.5 years after vaccination of 4-6 year olds with either DTPa or dTpa, the model predicted that 10 years post-vaccination, 98.6% of subjects vaccinated with dTpa were likely to remain seroprotected against diphtheria, compared to 99.6% vaccinated with DTPa. Therefore, the difference observed in diphtheria antibody geometric mean concentrations 1 month after booster vaccination at 4-6 years with dTpa or DTPa is unlikely to be of clinical relevance 10 years later at the time of the adolescent booster. PMID- 15530679 TI - Differential antibody responses to Plasmodium falciparum-derived B-cell epitopes induced by diepitope multiple antigen peptides (MAP) containing different T-cell epitopes. AB - Epitopes of universal character are needed when designing subunit vaccines against infectious diseases such as malaria. We have compared the immunogenicity of B-cell epitopes from the Plasmodium falciparum antigen repeats DPNANPNV (PfCS protein) and VTEEI (Pf332) when assembled with four different universal T-cell epitopes in diepitope multiple antigen peptides (MAP). T-epitopes employed were from P. falciparum antigens (CS.T3, [T(*)]4 and EBP3) or from the Clostridium tetani toxin (P2). In association with either of the T-epitopes, the genetic unresponsiveness to the B-epitopes was successfully bypassed. Our results show that the immunogenicity of a T-epitope alone does not necessarily predict the ability of the T-epitope to provide T-cell help when combined with other epitopes in an immunogen. Further, the nature of the immune responses in terms of total IgG antibodies and their subclass distribution, T-cell proliferation and IFN gamma production, varied with the T-epitope and mouse strain, which may indicate the need for inclusion of a combination of different universal T-epitopes in a future malaria subunit vaccine. PMID- 15530680 TI - Adult tetanus, diphtheria and pertussis immunization: knowledge, beliefs, behavior and anticipated uptake. AB - BACKGROUND: Lifetime protection against pertussis has been adopted as a goal of immunization programs in Canada. To anticipate adult coverage with a combined product containing tetanus (T) and diphtheria (d) toxoids and acellular pertussis (aP) vaccine as a booster dose, we conducted a survey of households in British Columbia, Canada. METHODS: In a random telephone survey involving 800 adults, 25 years of age and older, we assessed current behaviors related to adult Td immunization and beliefs regarding pertussis vaccine under various scenarios relevant to adult decision-making. RESULTS: Forty-five percent of participants reported having received tetanus vaccine within the previous 10 years; this rate was lowest amongst elderly persons 65 years of age or more (28%). On multi variate analysis, being up-to-date with tetanus immunization was independently associated with belief that an adult should be immunized against tetanus and perception that tetanus is life-threatening and inversely associated with being elderly. At baseline, 59% of respondents indicated willingness to receive pertussis immunization if provided free; this increased to 76% following sequential information about communicability and severity of pertussis illness and safety, efficacy and convenience of vaccine and up to 87% if accompanied by physician or nurse recommendation. Sixty-three percent of adults indicated they would receive the vaccine if required to pay $40.00 (Cdn) for it. CONCLUSIONS: Personal risk perception, public funding and physician recommendation are important to adults when considering tetanus and pertussis immunization. These factors may be relevant as immunization programs are expanded to include more adults generally. PMID- 15530681 TI - Influenza and pneumococcal vaccination as a model to assess C-reactive protein response to mild inflammation. AB - This study was set up to examine whether an influenza vaccine or an influenza vaccine in combination with pneumococcal vaccine can be used as a model to study responses to mild stimulation of the inflammatory system. In this study, 19 subjects received the influenza vaccine, 20 subjects the combination of influenza and pneumococcal vaccine. CRP and prothrombin fragment 1 and 2 (F1+2) were measured at baseline, and two times after vaccination. Influenza vaccination increased CRP by 0.20 mg/L, and influenza in combination with pneumococcal vaccine increased CRP by 0.60 mg/L. F1+2 increased 0.15 nmol/L after the combined vaccination; an increase in response to the influenza vaccination was not statistically significant. Our findings show that the influenza vaccine alone as well as the combination of the influenza and pneumococcal vaccine increases CRP levels with a peak 2 days after vaccination. PMID- 15530682 TI - High epitope density in a single recombinant protein molecule of the extracellular domain of influenza A virus M2 protein significantly enhances protective immunity. AB - The degree of epitope density has been shown to be a critical factor influencing the magnitude of epitope-specific responses. However, whether high epitope density in just a single protein molecule can still enhance the humoral response or, more importantly, the protective immunity, has not been determined. To test this, five glutathione-S-transferase fusion proteins bearing various numbers of copies of the M2e epitope on M2 protein of influenza virus (1, 2, 4, 8 and 16 copies) were prepared, and used to immunize mice and rabbits. Our data show clearly that M2e-specific humoral response was enhanced with increasing epitope density. By lethal challenge assay in mice, it was observed that recombinant proteins with higher M2e epitope densities resulted in higher survival rates and slower weight losses. The survival rate was directly related to the degree of epitope density in the single recombinant protein: 100% in the case of 16 M2e copies; 50% with 4 M2e epitopes; and 0% with one. PMID- 15530683 TI - Beliefs and behaviours: understanding chiropractors and immunization. AB - BACKGROUND: Concerns have been raised about the beliefs and behaviours of chiropractors related to immunization; however, none have systematically examined the relationships between beliefs and behaviours. PURPOSE: We examine the immunization-related behaviours and beliefs of chiropractors in Alberta, Canada, and explore the relationship of beliefs to immunization-related behaviours with patients. METHODS: Data were collected in 2002 from a postal survey of Alberta chiropractors. The questionnaire inquired about six behaviours of interest in the six months prior to survey (gave information about risks/benefits of vaccination; advised patients in favour/against have self/children immunized; counselled on freedom of choice; directed to sources of information on immunization). It included items addressing beliefs and norms related to immunization. RESULTS: The response rate was 78.2% (503/643). Immunization arose with patients at least monthly for 36.5% of respondents, and at least weekly for 9.2%. One quarter advised patients in favour and 27% against having themselves/their children immunized. A parsimonious model of chiropractor pro/anti-vaccination behaviours included beliefs about the efficacy/safety of vaccination, chiropractic philosophy and individual rights. CONCLUSIONS: Similar proportions of chiropractors advise patients in favour or against immunization. A small minority deals with immunization issues frequently. Behaviours can be understood in the context of beliefs. PMID- 15530684 TI - High levels of antibody in adults three years after vaccination with a reduced antigen content diphtheria-tetanus-acellular pertussis vaccine. AB - There is increasing interest in prevention of pertussis in adults by vaccination, but little is known about the duration of the antibody response to pertussis, diphtheria or tetanus in reduced antigen content vaccines formulated for adult use. Follow-up of a clinical trial including 550 adults comparing responses to reduced antigen content diphtheria-tetanus-acellular pertussis (dTpa) vaccine, or a licensed Td vaccine, provided the opportunity to evaluate this. Blood samples were collected at 0, 1, 12, 24 and 36 months following vaccination; of the original cohort of 550, 387 subjects (dTpa group N=310, Td+pa group N=77) were tested at month 36. Following a decrease in antibody levels against all vaccine antigens between one and 24 months following vaccination, levels stabilized during the third year, remaining higher at 36 months than pre-vaccination for all vaccine antigens. In particular, more than 90% of subjects remained seropositive for pertussis toxin and pertactin antibodies at 36 months after vaccination, suggesting ongoing protection against pertussis. Adult-formulated dTpa vaccines could replace Td for routine booster vaccination of older individuals. PMID- 15530685 TI - Cellular immune responses to feline immunodeficiency virus (FIV) induced by dual subtype FIV vaccine. AB - Vaccine-induced T cell responses to FIV were assessed by measuring FIV-specific cytokine and cytotoxic-effector molecule production. A total of 22 cats at 10-12 weeks of age received either dual-subtype FIV vaccine (n=12), uninfected cell lysate (n=5) consisting of cells used to produce vaccine viruses, or no immunization (n=5). Significant increases in mRNA and protein production of T helper 1 (Th1) cytokines (IL-2, IFNgamma), mRNA production of a cytotoxic effector molecule (perforin), and lymphoproliferation response were observed in peripheral blood mononuclear cells (PBMC) from dual-subtype FIV-vaccinated cats after in vitro stimulation with inactivated FIV. In contrast, no statistically significant increase in FIV-stimulated mRNA production of Th2 cytokines (IL-4, IL 6) or other cytotoxic-effector molecules (TNFalpha, FasL) was observed in the PBMC from dual-subtype vaccinated cats. Moreover, no FIV-specific increases in the IFNgamma, IL-2, and perforin mRNA productions and in the IFNgamma bioactivity and lymphoproliferation responses were observed in the PBMC from cell-immunized cats. These observations suggest that IFNgamma induction, lymphoproliferation, and significant portion of IL-2 and perforin productions in the PBMC from dual subtype vaccinated cats are clearly specific for viral antigens. Overall, dual subtype FIV vaccine elicited strong Th1 response (IFN(, IL-2), which may contribute to the vaccine protection by enhancing the perforin-mediated cytotoxic cell activity against FIV. PMID- 15530686 TI - Hepatitis B vaccination in a hyper-endemic tribal community from India: assessment after three years. AB - Hepatitis B infection is highly endemic among the primitive tribes of Andaman and Nicobar Islands, India and it is necessary to initiate hepatitis B vaccination for control of this infection. A pilot project of mass hepatitis B vaccination using indigenously developed vaccine was initiated among Nicobarese tribe of Car Nicobar Island. Sero-protection rates after second and third year were 89% and 85.5%, respectively. The rate of chronic infection in the vaccinated population after three years was 1.86% compared to the pre-vaccination rate of 20.7%. Considering high sero-protection rates and low cost, the indigenous vaccine could be used for vaccination programme in this tribal community. PMID- 15530687 TI - Expression of a vaccine protein in a plant cell line using a geminivirus-based replicon system. AB - Edible vaccines have been generated from both transgenic plants as well as from plant viral vectors. Here, we have taken the best attributions of both systems and designed a minimalized version of the bean yellow dwarf geminivirus (BeYDV) based replicon consisting of the cis-acting elements required for BeYDV replication as a means to express foreign genes in a plant cell line. Replication can be switched on at high levels upon expression of the BeYDV Rep protein, and gene expression enhanced enormously. Construction of an expression cassette encoding a synthetic vaccine gene and analysis of expression levels of a vaccine protein in a plant cell line system are described. PMID- 15530688 TI - Media coverage of anthrax vaccination refusal by Australian Defence Force personnel. AB - BACKGROUND: During February 2003 a number of Australian sailors were returned home from their deployment to the Persian Gulf after refusing anthrax vaccination. This paper examines the media coverage of this episode as a case study in how controversies about vaccine safety escalate. METHODS: Frame analysis of articles from major Australian newspapers (n=83) and transcripts of radio and television news and current affairs programs (n=22) to identify the main supportive and oppositional themes used in reportage and media debate. FINDINGS: Initially, the major news frames were supportive of the vaccine refusing soldiers, and conveyed a sense of distrust of the government's actions. These initial themes were rapidly re-framed and new dominant discourses appeared. First, sailors went from brave whistleblowers to being portrayed as deserters and cowards. Second, proponents shifted from their portrayal as faceless regulators to personal risk takers embodied in a well-respected Major General having the vaccine. Third, the voluntary nature of the vaccine was emphasised, thus dousing the flames of implied coercion. CONCLUSION: Marked shifts in the representation of vaccine opponents and proponents possibly contributed to the rapid diminishment of media interest in the story. PMID- 15530689 TI - Immune responses and protective efficacy in ponies immunised with an equine influenza ISCOM vaccine containing an 'American lineage' H3N8 virus. AB - Protective responses generated by vaccination with an immuno-stimulating complex (ISCOM)-based vaccine for equine influenza (EQUIP F), containing a new 'American lineage' H3N8 virus, were studied. Seven ponies in the vaccine group received two intramuscular injections of EQUIP F given 6 weeks apart. Aerosol challenge with an A/eq/Newmarket/1/93 reference strain 4 weeks after booster vaccination resulted in clinical signs of infection and viral shedding in 7 influenza-naive control animals whereas the vaccinated ponies were significantly protected from both clinical signs and virus excretion. Influenza virus-specific IgG responses in serum following immunisation with the ISCOM vaccine were predominantly of the IgGa and IgGb sub-isotypes, a pattern similar to that generated by equine influenza virus infection. However, in contrast to the response following infection, virus-specific antibody responses in nasal washes following immunisation were characterised by the presence of IgG but not IgA.These results demonstrated that an ISCOM-based vaccine containing A/eq/Kentucky/98 provides strong protective immunity against challenge with an 'American lineage' H3N8 reference virus. PMID- 15530690 TI - Immunogenicity and safety of a booster vaccination against tick-borne encephalitis more than 3 years following the last immunisation. AB - This study investigated the immune response and safety in 430 adults, when boosted more than 3 years after primary or booster TBE immunisation as measured by neutralization test (NT) and ELISA. Tested by NT, the post-booster day 21 geometric mean titer (GMT) was 331 and 142 for the 18-49 and > or =50 years old, respectively. The post-/pre-booster geometric mean titer ratio (GMR) was 2.29 for the 18-49 years old and 3.21 for the > or =50 years old. An at least four-fold increase of neutralizing TBE antibodies was observed in only 26 and 38% of subjects aged 18-49 and > or =50 years, respectively. The booster effect in subjects with only the primary vaccination course prior to study entry clearly depended on the time elapsed since last TBE vaccination with an estimated annual decline rate of 15%. In subjects with at least one additional booster vaccination virtually no antibody decline was observed. This study clearly indicates that (1) adults may be effectively and safely boosted with a different TBE vaccine and (2) following four immunisations protective antibodies can be detected far beyond a period of 3 years, thus, strongly supporting the reconsideration of currently recommended booster intervals. PMID- 15530691 TI - Comparison of antibodies directed against human respiratory syncytial virus antigens present in two commercial preparations of human immunoglobulins with different neutralizing activities. AB - Antibodies directed against human respiratory syncytial virus (HRSV) from two commercial preparations of human immunoglobulins (Igs) were compared. One of the Ig preparations (RespiGam) was obtained from blood samples selected for high titres of anti-HRSV neutralizing antibodies. The other preparation (Flebogamma) was obtained from unselected blood donations. RespiGam and Flebogamma had very similar anti-HRSV ELISA titres, but RespiGam neutralized virus infectivity 8-10 times more efficiently than Flebogamma. The same behaviour was observed when purified antibodies from RespiGam and Flebogamma, specific for either the fusion (F) or the attachment (G) glycoprotein, were compared. To gain further information about differences in neutralization between these two Ig preparations, antibodies recognizing certain F and G protein fragments or peptides were purified and their neutralizing activities were compared. In general, antibodies purified from RespiGam showed higher neutralizing activity that those purified from Flebogamma, but those differences were higher with antibodies specific for certain protein segments than for others. Some of the protein regions recognized by human neutralizing antibodies were mapped outside antigenic sites identified previously with panels of murine monoclonal antibodies. These results offer the possibility of searching for new neutralizing antibodies that could be used to study the molecular basis of neutralization and to prevent HRSV infections. PMID- 15530692 TI - Measles vaccination in the presence of maternal antibodies primes for a balanced humoral and cellular response to revaccination. AB - Early or low dose antigen exposure can prime the immune system for subsequent responses; the so-called "prime-boost" effect. In the context of a Sudanese measles vaccine trial, we assessed whether or not such early exposure could influence the response to revaccination. Children received either Connaught high titer vaccine (CN: n = 53; 10(4.7)pfu) or meningococcal A + C vaccine as a placebo (MEN: n = 58) at 5 months of age. At 9 months of age, all received standard titer Schwarz vaccine (SCH: 10(3.9)pfu). Neutralizing antibodies were measured before initial vaccination and at 9 months of age (plaque reduction neutralization assay (PRN)) and again at 5 years of age (syncytium inhibition assay (SIA)). Lymphoproliferative responses to measles virus (MV) antigens were evaluated at 5 years of age. Eleven of the 53 CN-SCH children (21%) had sub protective neutralizing antibody titers prior to revaccination (log PRN 1.5 +/- 0.03 versus 2.9 +/- 0.07 in the remaining 42 children; P < 0.004). Maternal antibody titers at the time of initial vaccination in these 11 were high (PRN 2.44 +/- 0.12 versus 1.9 +/- 0.04; P < 0.0001). At 5 years of age, neutralizing antibodies were comparable in the 11 CN-SCH poor responders (log SIA 2.1 +/- 0.09), the remaining CN-SCH children (2.2 +/- 0.06) and the MEN-SCH group vaccinated only once at 9 months of age (2.25 +/- 0.06). In contrast, 7/11 of the CN-SCH poor responders (64%) had stimulation indices (SI) > 3 in response to MV antigens at 5 years of age (SI 3.1 +/- 0.6) compared with only 14% in the remaining children of the CN-SCH group (2.0 +/- 0.3; P = 0.05) and 8% in the MEN SCH group (1.4 +/- 0.2; P < 0.0003). These data suggest that early measles vaccination in the presence of maternal antibodies can sometimes prime for a balanced humoral and cellular immune response to subsequent revaccination. PMID- 15530693 TI - Oral immunization of Carassius auratus with modified recombinant A-layer proteins entrapped in alginate beads. AB - This study was focused on the utilization of a recombinant expression system to produce a unique modified subunit vaccine possessing a self-contained delivery system which could potentially improve the uptake and delivery of vaccine products as well their immunogenic potential. For this purpose the A-layer protein (At-R) associated with the fish pathogen atypical Aeromonas salmonicida was cloned and modified by the genetic fusion of the protein transduction domain (MTS) derived from Kaposi fibroblast growth factor (At-MTS). The potential for these proteins to be employed as antigens for oral immunization of goldfish was examined by encapsulation of At-R, At-MTS and the control, BSA, into biodegradable alginate gel macrospheres which were fed to goldfish in place of standard pellet fish feed. The bead physical properties were modified only in the presence of At-R and the temporal release of proteins was significantly less when At-MTS was employed. Western blot analysis of serum samples collected from fish following intubation with the recombinant proteins determined that the rate of protein uptake from the digestive tract into the blood system improved considerably when MTS was fused to At-R. Experimental fish were fed one of three protein-alginate formulae on a schedule of 3 days/week or 5 days/month for a period of 2 months. After 1 month, animals fed on the 5-day protocol demonstrated increased serum antibody titers while following an additional month of feeding this level decreased and titers were found to be higher in fish maintained on the 3-day regime. Fish fed At-MTS maintained the highest titer at the end of 2-month period. To determine whether the diminished antibody titers were a result of oral tolerance fish were injected intraperitoneally with the At-R antigen. Only experimental groups which had been fed At-R or At-MTS demonstrated increased antibody titers which paralleled a typical secondary humoral response. In spite of the presence of an increased titer to A-protein, vaccinated fish did not demonstrate resistance to infection with atypical A. salmonicida. PMID- 15530694 TI - Cationic polymers that enhance the performance of HbsAg DNA in vivo. AB - In this paper, different cationic polymers were investigated as a DNA delivery system both in vitro in dendritic and muscle cells and in vivo, in a murine model. Expression of the reporter gene beta-galactosidase was used in order to determine the in vitro transfection efficiency of these polymer-DNA complexes (polyplexes) and both specific mRNA and protein expression were monitored in parallel with polyplex toxicity on the cells. Interestingly, the enhancing expression activities of the different polyplexes were tissue-dependent, implying that they may gain entrance to the cells through specific receptors. Subsequently, complexes of polymers and DNA plasmid (pCMV-S) encoding the human hepatitis B virus (HBV) surface antigen (HBsAg) were injected into the skeletal muscles of BALB/c mice. Higher levels of both HBsAg local expression in the tibial anterior muscles and systemic humoral immune responses were detected when the selected polymers complexed with pCMV-S were compared to those complexed with pCMV-S alone. Induction of immunoglobulin G2a (IgG2a) against HbsAg in the serum of pCMV-S-polyplex vaccinated mice varied with the polymer used, suggesting that polyplex-mediated DNA vaccination can potentially modulate the type of helper T cell immunity (Th). The effect of some polyplexes to switch the host immune response more towards a Th1 response may be associated with their differential efficiency to transfect dendritic cells and/or other antigen-presenting cells (APC) as was observed in vitro. These results suggest that the investigated cationic polymers can be effective as delivery/adjuvant compounds for DNA. PMID- 15530695 TI - Economic evaluation of meningococcal serogroup C conjugate vaccination programmes in The Netherlands and its impact on decision-making. AB - The cost-effectiveness of one time vaccination of all persons aged 14 months to 18 years (catch-up programme) and of routine childhood immunisation at either ages 2 + 3 + 4 months, 5 + 6 months, or 14 months with a meningococcal C conjugate vaccine was estimated for The Netherlands, from a societal and a health care payer perspective. A decision analysis cohort model was employed (time horizon 77 years), direct and indirect costs (friction cost method) were considered and future costs and effects were discounted at 4%. The results showed that all vaccination options yield a substantial health gain and that the catch up programme and routine vaccination at 14 months render favourable cost effectiveness ratios: between about 13,200 and 17,700 per life year gained for the catch-up programme and between about 2200 and 2400 per life year gained for routine childhood vaccination at 14 months, depending on the perspective. In comparison to vaccination at 14 months, routine childhood vaccination during the first year of life is much less cost-effective: each additional life year gained costs approximately 147,000 (2 + 3 + 4 months) or 102,000 (5 + 6 months), from both perspectives. Additionally, inclusion of the likely herd immunity effect of the catch-up programme increases these incremental cost-effectiveness ratios. These results played a major role in the decision to add meningococcal C vaccination to the routine childhood immunisation schedule at 14 months and to implement a catch-up vaccination programme in The Netherlands in 2002. PMID- 15530696 TI - A recombinant rubella virus E1 glycoprotein as a rubella vaccine candidate. AB - A recombinant rubella virus E1 (rE1) glycoprotein was produced and some of its chemical and immunological features were characterized. Two animal models were then used to establish that the rE1 glycoprotein and rubella virus particles shared antigenic and immunogenic properties. In the first one, sera from rE1 glycoprotein-immunized BALB/c mice neutralized in vitro rubella virus infection. In the second model, severe combined immune deficient (SCID) mice implanted with tonsil fragments from rubella immune donors and immunized with rE1 glycoprotein produced human anti-rubella virus antibodies. Altogether, these results showed that immunization with rE1 glycoprotein elicited neutralizing anti-rubella virus antibodies. This study thus indicated that the rE1 glycoprotein could constitute a non-replicating rubella vaccine. PMID- 15530697 TI - Nasal immunization of mice with a rotavirus DNA vaccine that induces protective intestinal IgA antibodies. AB - DNA vaccination using a plasmid encoding the rotavirus inner capsid VP6 has been explored in the mouse model of rotavirus infection. BALB/c mice were immunized with a VP6 DNA vaccine by the intramuscular, nasal and oral routes. VP6 DNA vaccination by the nasal and oral routes induced the production of anti-VP6 IgA antibodies by intestinal lymphoid cells. Intramuscular DNA injection stimulated the production of serum anti-VP6 IgG but not serum anti-VP6 IgA antibodies. Protection against shedding of rotaviruses in stools after oral challenge with the murine EDIM rotavirus strain was investigated in the immunized mice. A significant reduction in the level of rotavirus antigen shedding was demonstrated in those mice immunized at mucosal surfaces, both orally and nasally, with the VP6 DNA vaccine. Intramuscular DNA immunization, which elicited serum anti-VP6 IgG responses but not virus-specific intestinal IgA antibodies, did not provide significant protection against rotavirus challenge. PMID- 15530698 TI - Recombinant viruses obtained from co-infection in vitro with a live vaccinia vectored influenza vaccine and a naturally occurring cowpox virus display different plaque phenotypes and loss of the transgene. AB - Some poxviruses are very attractive as transgenic vaccine vectors for humans, domestic animals and wildlife. Poxviridae family members circulate in different ecosystems and parts of the world, providing a pool of possible recombination partners for released or escaped genetically modified poxviruses. We performed in vitro double infections with a vaccinia virus strain Ankara (MVA) vectored influenza vaccine and a cowpox virus isolate from Norway, isolated hybrids, and further analyzed three hybrid viruses with different plaque phenotypes. One of the hybrids was genetically unstable, and during adaptation to new host cells its MVA derived influenza gene was deleted at a high frequency. This is significant in a risk assessment context, since the transgene would be the only logical tag for monitoring unwanted spread and non-target effects of a vaccine virus. Putative recombination events involving genetically modified and naturally occurring viruses should be included in health and environmental risk assessments. PMID- 15530699 TI - Detection of cytomegalovirus infection during clinical trials of glycoprotein B vaccine. AB - Preabsorption of sera with cytomegalovirus (CMV) glycoprotein B (gB) prior to testing for CMV IgG antibody was evaluated for detection of CMV infection during CMV gB vaccine clinical trials. Although, 98.2% of 109 CMV gB vaccine recipients were seropositive with a standard assay, preabsorption of sera with gB reduced their CMV antibody to extremely low levels compared with subjects with CMV infection. One subject who acquired CMV during a vaccine trial was easily identified by gB-absorbed CMV IgG results. The gB-absorbed CMV IgG antibody assay is a promising approach for screening CMV gB vaccine trial participants for CMV infection. PMID- 15530700 TI - Vero-cell rabies vaccine produced using serum-free medium. AB - A new rabies vaccine was developed from Vero cells adhered to microcarriers, cultivated in a bioreactor in serum-free medium and infected with the PV/VERO Paris rabies virus strain. The viral suspensions were concentrated by tangential filtration, purified by chromatography and inactivated with beta-propiolactone. In immunogenicity studies performed in mice immunized with three doses of the new vaccine (seven batches) and the commercial Verorab and HDCV, mean titers of neutralizing antibodies of 10.3-34.6, 6.54 and 9.36 IU/ml were found, respectively. The vaccine presented stability during 14 months at 2-8 degrees C, 30 days at 37 degrees C and 8 h at 45 degrees C. The use of serum-free medium facilitated the downstream process leading to residual cellular DNA values <22.8 pg per dose of vaccine in all produced batches. The effective immunogenicity induced in mice by this vaccine, the degree of purity of the product, the high antigen yield and the reduction of the cost of the product due to the virus production and purification processes, makes this technology very important for countries where rabies presents a great public health problem. PMID- 15530702 TI - Long-term health effects of repeated exposure to multiple vaccines. AB - The health of 155 former workers in a US military research program who had received multiple vaccines and 265 matched community controls was assessed. The study population was mostly male (83%) and elderly (median age, 69 years). Multiply immunized (MIP) subjects received vaccines and/or skin tests (median = 154) over a median of 17.3 years; interval from start of immunizations to survey completion was 15-55 years (mean = 43.1 years). MIP subjects characterized themselves as slightly less healthy than controls (P = 0.057). Fatigue (but no other symptom) was reported more frequently in the MIP group (P = 0.011), but was not associated with number of injections, number of vaccines, or time in program. No differences between MIP and control groups were seen for numerous self reported medical conditions. Several statistically significant abnormalities were seen in clinical laboratory tests among MIP subjects, but none appeared to be clinically significant. A significant difference in frequency of monoclonal spikes and/or paraprotein peaks between MIP (12.5%) and control (4.5%) groups (RR = 2.7, P < 0.003) was observed; no associations with lifestyle, vaccine exposure, or medical conditions were found. PMID- 15530701 TI - In vitro growth and stability of recombinant rabies viruses designed for vaccination of wildlife. AB - Three live rabies virus (RV) recombinant vaccine candidates, SPBNGA, SPBNGA-Cyto c (+), and SPBNGA-GA, were examined for their production levels and stability. Maximum production levels up to 10(10) infectious particles/mL were achieved using bioreactor technology. All virus lots exhibited thermostability profiles typical for RV vaccines and were non-pathogenic for intracranially inoculated immunocompetent mice. Moreover, sequence analysis indicated high genetic stability in all three RVs during 10 consecutive passages in newborn mice. This analysis revealed no change in the extra RV G gene in the SPBNGA-GA vaccine or in the cytochrome c gene in the SPBNGA-Cyto c (+) vaccine. Moreover, no changes were detected in the G gene codon for Glu333, which renders the virus non-pathogenic. However, after the fifth passage, a mutation resulting in an Asn194 --> Lys194 exchange emerged in the G genes of all three RVs. This mutation was associated with a modest increase in pathogenicity in SPBNGA and SPBNGA-Cyto c (+), but not in SPBNGA-GA, which contained the mutation in only one of its two G genes and which remained non-pathogenic. These results demonstrate the feasibility of producing RV vaccines that remain highly stable even after multiple passages. PMID- 15530704 TI - Differential polarization of immune responses by co-administration of antigens with chemokines. AB - Chemokines are key players in the elicitation of immune response, by selectively attracting subpopulations of immune cells to the site of antigen presentation. Therefore, they are natural candidates for modulating immune responses to antigens qualitatively and quantitatively. We have selected chemokines associated with different arms of the immune response, i.e. RANTES/CCL5, B-lymphocyte chemoattractant/CXCL13, and monocyte chemoattractant protein-1/CCL2, and co injected DNA expression constructs encoding these chemokines with constructs encoding two HIV antigens, gp120 and gp160, in mice. We subsequently measured markers of both cellular and humoral immune responses, and found that these chemokines qualitatively influenced the outcome of immune responses to both antigens, essentially according to their predicted association to Th profiles. These results are relevant towards the engineering of novel vaccine and immune based therapies, and point to chemokines as candidate adjuvant and immunomodulatory molecules. PMID- 15530703 TI - Prevention of naturally occurring infectious bovine keratoconjunctivitis with a recombinant Moraxella bovis cytotoxin-ISCOM matrix adjuvanted vaccine. AB - The efficacy of a recombinant Moraxella bovis cytotoxin subunit vaccine to prevent naturally occurring infectious bovine keratoconjunctivitis (IBK) was evaluated in a randomized, blinded, controlled field trial. Ninety-three cross bred beef calves were vaccinated with either saline, ISCOM matrix (adjuvant control), or a recombinant M. bovis cytotoxin carboxy terminus peptide plus ISCOM matrix and boostered 21 days later. Ocular examinations were performed once weekly for 20 weeks. At week 12, the cumulative proportion of calves with ulcerated eyes in the recombinant vaccine group was significantly lower than in the saline control group. Throughout the 20 week trial, the cumulative proportion of ulcerated calves remained lowest in the recombinant vaccine group. By week 7, nonulcerated calves in the recombinant vaccine group had significantly higher changes in serum neutralizing titers and cytotoxin specific to total IgG ratios in serum and tears as compared to calves in the control groups. The trend for a reduced cumulative proportion of IBK in the vaccinated calves over the 20 week trial suggests that a recombinant M. bovis cytotoxin vaccine may be beneficial in helping to prevent naturally occurring IBK. PMID- 15530705 TI - A two-codon mutant of cholera toxin lacking ADP-ribosylating activity functions as an effective adjuvant for eliciting mucosal and systemic cellular immune responses to peptide antigens. AB - Vaccination with peptide antigens is an effective strategy against mucosal viral infections. We tested a two-codon mutant of cholera toxin (CT-2*) lacking ADP ribosylating activity and toxicity as a mucosal adjuvant for T cell epitope peptides for intranasal immunization of mice. Efficient induction of helper and cytotoxic T lymphocyte responses associated with TH1 cytokine production were observed in the systemic and mucosal compartments including nasal, gut, and vaginal associated lymphoid tissues. Single or multiple dosing with the peptide antigen and CT-2* induced strong memory immunity without tolerance. These results demonstrate CT-2* as a suitable mucosal adjuvant for priming antigen-specific cellular immune responses. PMID- 15530706 TI - Management of preterm labour and delivery: the future. PMID- 15530707 TI - What is the future of tocolysis? AB - Use of the currently available tocolytics is controversial because it has not been associated with improved perinatal outcomes. New markers of preterm labour may come from gene-profiling studies, in as much as they may help in identifying novel genes regulating myometrial quiescence and in expanding our understanding of the pathologic process of uterine dysfunction. Study of certain transcripts in circulating white blood cells by RT-PCR could assist the obstetrician in evaluation of the risks. Uterine electromyography (EMG) also has the potential benefit of monitoring tocolytic treatment, although no standard method of clinical interpretation has yet been devised for the results yielded by this instrumentation. Recent functional genomic studies found that in the uterus at term there is a massive down-regulation of a large panel of developmental cell adhesion molecules and proliferation-related genes. Conversely, maintaining the developmental processes in an active state in patients at risk would help to prevent preterm delivery. It is too early to suggest any therapies with anticytokines in pregnant women. However, exploration of genetic polymorphisms, which may influence the balance of pro- and anti-inflammatory cytokines that are relevant to the course of preterm labour, seems to be a novel avenue that should be explored. PMID- 15530708 TI - Cyclic nucleotide phosphodiesterase-4 inhibitors: a promising therapeutic approach to premature birth? AB - Pharmacological interventions in preterm labour attempt to target various protagonists involved in the signalling pathway leading to myometrial contractions. None of these interventions has clearly demonstrated any real clinical benefit, and some are associated with severe maternal and fetal side effects. The original study now reported has disclosed a new potential target, myometrial phosphodiesterase-4 (PDE4), for tocolysis. The PDE4 family specifically hydrolyses cAMP, a major intracellular second messenger implicated in the mechanisms governing smooth muscle motility. One particular isoform, PDE4B2, is overexpressed in the human myometrium at the end of pregnancy, and we observed a change of the myorelaxant properties of selective PDE4 inhibitors at this time. Signalling factors involved in labour at term or in infection-induced preterm labour, such as PGE2 or IL-1beta, are also able to induce PDE4B2 expression by way of a cAMP-dependent pathway. This feedback loop explains, at least in part, the process of desensitisation to relaxant agents that occurs at the end of pregnancy. In addition, PDE4 inhibitors block the production of LPS induced inflammatory cytokines by myometrial explants. Thus, a combination of myorelaxant and anti-inflammatory properties make the PDE4 inhibitors particularly attractive as a target for the prevention of threatened infection induced preterm delivery and its consequences for the premature infant. PMID- 15530709 TI - Is there a place for inhibition of cervical ripening in the management of preterm labour? AB - Although premature cervical ripening is one of the two major pathophysiological processes occurring in preterm labour, research has mostly focused on the development of tocolytic agents. Our goal in this study was to review the recent progress made in the understanding of cervical ripening physiology and to explore the molecules that could be targeted for the development of new pharmacological compounds capable of inhibiting this process and to further improve the management of preterm labour. PMID- 15530710 TI - Perspectives in the prevention of premature birth. AB - Obstetric and neonatal interventions have improved the survival of preterm infants, but there has not been an equivalent reduction in long-term neurological disability. Thus, some effort must be invested in finding ways of preventing preterm birth. Numerous programmes have been promoted to address the matter of how the frequency of preterm birth could be prevented. Most interventions intended to prevent preterm labour do not have the desired effect, except for antibiotic treatment in cases of asymptomatic bacteriuria or bacterial vaginosis and progesterone administered prophylactically in high-risk women. Tocolytic drugs appear to delay delivery long enough for successful administration of corticosteroids in women in preterm labour, but without decreasing the risk of preterm birth. Some authors promote public health approaches that address all risk factors and affect the entire population of pregnant women, given that prevention programmes directed only at high-risk women have had little effect in preventing preterm births. However, the lack of progress in reducing the frequency of preterm births is also due to our limited understanding of the aetiology of preterm delivery. Although there is growing evidence that infection and neuroendocrine processes are involved, progress has remained slow. Recently, the hypothesis of a genetic predisposition to preterm delivery has been set up. Additional research exploring the pathophysiology of preterm labour is obviously needed, which will hopefully lead to the development of new therapeutic approaches. PMID- 15530711 TI - Could the consequences of premature delivery be further attenuated by means of new prenatal strategies? AB - The last 10 years have already seen improvements in the short- and long-term prognosis for premature neonates. Nevertheless, progress in the prenatal evaluation of predictive factors for neonatal diseases and more detailed and reliable knowledge of fetal physiology could allow the development of new treatments with consequent expectations of further improvements in the prognosis for such premature newborns. Global strategies for the management of preterm labour, ranging from a policy for prenatal transfer to centres offering the appropriate level of perinatal care, should continue to be expanded, and long term evaluations must also be continued. PMID- 15530712 TI - Can obstetric care provide further improvement in the outcome of preterm infants? AB - Reducing the incidence of cerebral damage in preterm infants has become a major objective of perinatal medicine. Recent studies have shown that intrapartum hypoxia is implicated in only 10% of cases, whereas prenatal factors are significantly linked with such damage. The main risk factors associated with cerebral palsy are preterm birth, multiple pregnancy, intrauterine infection, serious hypoxaemic and haemodynamic disorders and, possibly, thrombophilic disorders. Recent progress in recognition of the pathogenesis of cerebral white matter damage has underlined the roles of cytokines, including interleukins 1 and 6 and tumour necrosis factor alpha (TNFalpha), and of a massive release of glutamate, which leads to the excitotoxic cascade. Three measures have had demonstrable benefits in improving neonatal outcome in preterm infants: a policy of prenatal transfers to tertiary level care units, antenatal corticosteroid therapy, and administration of antibiotics to women with preterm premature rupture of membranes. The influence of tocolysis is usually considered to be relatively minor, but further studies in very preterm pregnancies may be required. The recent development of tocolytics without major adverse effects will probably facilitate such studies. The mode of delivery of early preterm infants is another important area of controversy. Recent studies have suggested that a policy of elective caesarean section in PPROM and in the case of breech presentation of fetuses weighing less than 1000-1500 g would be beneficial. Lastly, in vitro and animal studies have shown that several pharmacological agents can prevent white matter disease by interacting with cytokine and the excitotoxic cascade. This will probably constitute an important area of research in the future. PMID- 15530713 TI - Care of very premature infants: looking to the future. AB - Advances in prenatal care have improved survival rates in extremely preterm newborns, but cerebral palsy rates have not decreased in developed countries over the past 30 years. During the next 10 years we will probably not observe a dramatic improvement in intensive care such as that observed over the last 15 years. The man goal for the coming years will be to improve the quality of neonatal and postdischarge care in order to improve the long-term outcomes of very preterm infants. PMID- 15530714 TI - Ethical dilemmas in extreme prematurity: recent answers; more questions. AB - Advances in perinatal care allow survival of more extremely premature infants, but the implementation and continuation of intensive care may itself constitute an ethical dilemma, given the limited chances of intact survival among the patients most at risk. This paper discusses several key issues raised by the options that are under general consideration with reference to births of infants at the threshold of viability, in particular: the implications of making a distinction between extreme prematurity and other general medical situations that may involve decisions on ending support; the concrete nature of the restrictions on therapy in such patients interactions and the need for feedback between parents, medical staff and society. PMID- 15530715 TI - Is it possible to improve diagnostic and prognostic criteria of preterm labour? AB - Preterm labour is a frequent cause of admission to hospital during pregnancy. However, in most cases, preterm labour is diagnosed with reference to clinical criteria only. Since the clinical assessment of uterine contractions and of cervical changes is highly subjective, few of the patients admitted to the hospital with suspected preterm labour will ultimately deliver preterm. There is a need for sensitive methods of detecting patients who are genuinely at high risk of preterm birth, but on the other hand, specific methods of reducing unnecessary treatments or hospital admissions are also required. Recently, a few techniques such as ultrasonographic measurement of cervical length and fetal fibronectin have been introduced into clinical practice with the aim of improving prediction of the risk of actual preterm delivery. In the future, the assessment of cervical status may be based not only on anatomical changes, but also on functional criteria. New techniques are being developed for evaluation of the mechanical properties of the cervix (cervical distensibility), noninvasive measurement of its collagen content (light-induced fluorescence of cervical collagen), or even direct assessment of the changes in cervical water content (magnetic resonance imaging). Correlations have been found between these measurements and the risk of preterm birth, but clinical studies are still needed to allow better assessment of the predictive value of these new methods in clinical practice. PMID- 15530716 TI - Context of relapse for substance-dependent adults with and without comorbid psychiatric disorders. AB - OBJECTIVE: This study examined the context of initial posttreatment substance use episodes and 1-year treatment outcomes among substance abusers in relation to Axis I psychiatric diagnoses. METHOD: Of the 210 adults meeting DSM-III-R diagnostic criteria for a substance use disorder (SUD), 102 also met criteria for mood disorder, posttraumatic stress disorder (PTSD), or both (SUD-PSY). Quarterly face-to-face interviews and monthly telephone contacts were conducted following treatment to ascertain outcomes. RESULTS: Antecedents of initial posttreatment substance use differed for SUD adults compared to SUD-PSY. Negative affective states preceded posttreatment use episodes more frequently among SUD-PSY participants. The majority of SUD-PSY adults reported intrapersonal/environmental antecedents whereas the majority of SUD adults reported interpersonal antecedents. Compared to SUD adults without an Axis I disorder, the SUD-PSY group was more likely to resume substance use when alone. Participants drank more than twice as much alcohol when relapsing alone compared to those who resumed use with others. Negative affective state prior to initial use was predictive of more subsequent using days for comorbid adults but not SUD adults. CONCLUSIONS: These findings highlight differences in the process of resuming substance use for substance abusers with concomitant psychiatric disorders compared to SUD adults. Treatment implications are discussed. PMID- 15530718 TI - Adolescents' responses to the gender valence of cigarette advertising imagery: the role of affect and the self-concept. AB - The studies presented in this manuscript evaluated the role that affect and the self-concept play in adolescent never smokers' reactions to the gender valence of cigarette advertising imagery. Study 1 (n=29; 59% female) revealed that adolescent females have more positive affective reactions to female-valenced cigarette advertising imagery compared to male-valenced cigarette advertising imagery. Study 2 (n=101; 56% female) revealed that adolescent females viewed female-valenced cigarette advertising imagery as more relevant to their self concepts compared to male-valenced cigarette advertising imagery. Across both studies, male adolescents did not respond differently as a function of the gender valence of cigarette advertising imagery. Thus, female-valenced cigarette advertising imagery may have specific effects on never smoking female adolescents by enhancing positive affect and suggesting that women who smoke hold the same characteristics as do the young women themselves. PMID- 15530717 TI - Project EX--a teen smoking cessation initial study in Wuhan, China. AB - The increasing smoking prevalence in China indicates a need for effective smoking cessation programs, yet, to our knowledge, no studies have evaluated the effects of smoking cessation programs among Chinese adolescents. A group of 46 10th-grade level cigarette smokers from two schools in Wuhan, China, were provided with Project EX, an eight-session school-based clinic smoking cessation program developed in the United States. Efforts of translation of the Project EX curriculum, verification of translation, curriculum modification, and cultural adaptation were made to adapt the curriculum to the local culture. The 46 smokers represented 71% of all the self-reported 30-day smokers among 622 10th graders at these two schools. Only one student dropped out from the clinic program. Four month follow-up data indicated a 10.5% 30-day quit rate and a 14.3% 7-day quit rate. The students who did not quit smoking reported a 16% reduction in daily cigarette consumption at posttest and a 33% reduction at 4-month follow-up. Use of a 2 1/2-week prebaseline-to-baseline clinic assessment indicated a clinic cohort nonassisted quit rate of 3%. These data provided evidence that Project EX can be adapted in another country, such as China; can be very well received; and can lead to promising results on cessation. PMID- 15530719 TI - Development and validity of drinking pattern classification: binge, episodic, sporadic, and steady drinkers in treatment for alcohol problems. AB - This study refines an empirically derived drinking pattern classification system [Addict. Behav. 20 (1995) 23] and assesses its concurrent and predictive validity in a new sample of alcohol-dependent adults in treatment. Drinking data were collected from 195 adults (133 men) at baseline and for 52 weeks postbaseline using the Timeline Follow-back (TLFB) method. Ninety-three percent of the sample were classified into one of four drinking patterns: binge (n=13, 6.5%), episodic (n=41, 21%), sporadic (n=17, 9%), or steady (n=111, 57%). The steady drinking group showed substantial variability in drinking intensity and was divided into steady/high intensity (n=67, 34%) and steady/low intensity (n=44, 23%) subgroups. With age and gender controlled, the five subgroups did not differ on baseline employment or marital status, but differed on a measure of relationship functioning. Binge and steady/high groups reported the most severe alcohol-use histories. Steady/low intensity drinkers had later age of onset of alcohol problems. The five-pattern classification demonstrated predictive validity; steady (heavy and low intensity) drinkers most dramatically reduced the frequency of their drinking at 26 weeks postbaseline, while binge drinkers reduced their drinking intensity. Binge and steady/heavy drinkers were more likely than steady/low and sporadic drinkers to be abstinent at follow up. The final classification into five pattern groups shows promise of clinical utility for treatment-seeking samples. PMID- 15530720 TI - The influence of offering free transdermal nicotine patches on quit rates in a local health department's smoking cessation program. AB - Nicotine replacement therapy (NRT) has added to the menu of options available to assist cigarette smokers in quit attempts, but cost remains a barrier to access. A quasi-experimental study was carried out to compare quit rates and continuous abstinence from smoking before (n=601) and after (n=311) free nicotine patches were offered to smokers who participated in the Washington County (Maryland) Health Department's "Stop Smoking for Life" group behavioral cessation program. After free NRT was offered, the quit rates upon completion of the program increased from 38% to 65% [difference 27%; 95% confidence limits (CL) 21%, 34%]. The difference in continuos abstinence from smoking between the two groups was no longer statistically significant after 6 months of follow-up, reflecting the more rapid rate of reversion to smoking that occurred during the 18-month follow-up period among the free NRT group who had quit [adjusted rate ratio (RR) 1.35; 95% CL 1.03, 1.78]. Enrollment during the first 18 months after free NRT was 37% greater than the program's first 18 months (P=.08). In conclusion, adding free nicotine patches to a smoking cessation program was associated with increased program enrollment and significantly increased short-term-but not long-term-quit rates. The rapid reversion to smoking in the group who received free nicotine patches could potentially be obviated if participants extend their use of nicotine patches after the free 6-week supply is exhausted. PMID- 15530721 TI - Perceived family support, negative mood regulation expectancies, coping, and adolescent alcohol use: evidence of mediation and moderation effects. AB - The stressor vulnerability model of alcohol use posits that individuals learn to use drinking as a coping response when they believe healthier means of coping are unavailable. The present study evaluated hypotheses derived from the model adding two potential coping resources that may be important for adolescents--perceived family support and expectancies for negative mood regulation. Students in Grades 9-12 (N=210, 50% female) completed self-report measures of family environment, alcohol expectancies, coping preferences, negative mood regulation, motives for drinking, and drinking behavior. Recent drinking, lifetime drinking, and drunkenness were positively associated with stronger tension reduction alcohol expectancies, and drinking to cope (DTC) completely mediated these relations. DTC also was associated with avoidant coping dispositions and with the interaction between tension reduction alcohol expectancies and avoidant coping dispositions. Family support and NMR expectancies were only modestly correlated with DTC and not correlated with drinking measures. In regression analyses, family support and NMR expectancies were not associated with DTC independent of tension reduction expectancies and avoidant coping disposition. However, these two coping resource variables moderated the relations of other predictors with DTC and with drunkenness. The present study supports the stressor vulnerability model and highlights the importance of family support and NMR expectancies as coping resources related to drinking processes. Because many of the tenets of the model appear to extend to adolescents, it may have considerable utility as a conceptual framework for the design and implementation of prevention programs. PMID- 15530722 TI - Cigar use before and after a cigarette excise tax increase in New Jersey. AB - OBJECTIVE: To determine whether cigarette smokers in New Jersey substituted cigars following a cigarette excise tax increase. METHODS: We examined data from New Jersey's 2001 and 2002 Adult Tobacco Survey (NJATS). Adjusted odds ratios were generated for ever and current cigar use, controlling for demographics and cigarette smoking status. RESULTS: Males, whites, ever cigarette smokers, and those with higher education reported greater ever and current cigar use in both years. The adjusted odds for cigar use among recent quitters were higher in 2002 than in 2001. CONCLUSIONS: When other tobacco products' excise taxes fail to keep pace with cigarettes, there is a real potential for product substitution. Cessation of all tobacco products is necessary for the best public health outcome. PMID- 15530723 TI - Older, medically ill smokers are concerned about weight gain after quitting smoking. AB - Concern about postcessation weight gain has been shown to be a barrier to quitting for healthy smokers, but no study has examined these concerns among medically ill smokers. We examined whether medically ill smokers (N=271; 54% female, mean age=57 years) receiving nurse-delivered home health care report postcessation weight concern as a barrier to quitting smoking. Higher levels of weight concern were associated with younger age, longer duration of home care service, greater motivation to quit smoking, lower self-efficacy to quit smoking, more favorable views of smoking, and lower levels of social support; collectively accounting for over 20% of the variance. Increased illness severity was also associated with higher postcessation weight concern. Heavier smokers had higher levels of weight concerns at all follow-ups. Weight concerns did not prospectively predict smoking status, but quitters had higher subsequent levels of weight concerns than continued smokers. These results suggest that, even in the context of poor health and limited physical functioning, smokers are concerned about weight gain after quitting. PMID- 15530724 TI - Knowledge, attitudes, and practices of argileh (water pipe or hubble-bubble) and cigarette smoking among pregnant women in Lebanon. AB - BACKGROUND: Currently, little is known about argileh (water pipe or hubble bubble) and cigarette smoking among pregnant women in the Arab world, despite emerging evidence on the adverse health effects of argileh smoking and well established knowledge about the health risks of cigarette smoking during pregnancy. OBJECTIVES: The present study assesses pregnant Arab women's knowledge of chemical contents and related harmful effects of argileh and cigarettes, their attitudes towards smoking argileh and cigarettes, and their actual smoking of argileh and cigarettes, both before and during pregnancy. METHODS: A stratified sample of 864 women from 23 health care centers in Lebanon completed a structured, interviewer-administered questionnaire. Information was collected on basic demographic variables, women's knowledge, attitudes, and cigarette and argileh smoking. RESULTS: Women were partially knowledgeable about the health risks of cigarette smoking, knew little about the harmful ingredients of argileh smoking, and had many misconceptions regarding how argileh worked or how it can produce harm. Attitudes were permissive towards all forms of smoking. Almost one quarter (23%) of participants reported smoking during pregnancy, with 17% smoking only cigarettes, 4% smoking only argileh, and 1.5% smoking both cigarette and argileh. CONCLUSION: A significant and growing percentage of pregnant Arab women are smoking in Lebanon, with four cigarette smokers for every argileh smoker. Smoking behaviors are empirically linked with important gaps in knowledge and with permissive attitudes. These data may be used to design more effective prevention programs targeting this vulnerable population. PMID- 15530725 TI - Therapeutic communities for drug addicts: prediction of long-term outcomes. AB - The study, conducted among 167 heroin addicts from three therapeutic communities (TCs) in Israel, examined their drug use 15 months after their departure from the community and the contribution of sociodemographic characteristics, time in community, psychopathology, self-esteem, and locus of control to successful outcome. The findings show that about half the clients were clean at follow-up, that over 90% of those who completed the program were clean, and that the longer the client stayed in the TC, the more likely he or she was to be clean later. Drug use at follow-up was positively associated with prior criminal activity and negatively associated with living with a partner before entering the TC. In addition, high self-esteem and an independent locus of control (not believing that one's life was controlled by powerful others) were also associated with being clean at follow-up. PMID- 15530726 TI - Motivation to stop substance use and psychological and environmental characteristics of homeless women. AB - Characteristics associated with wanting to permanently quit their alcohol, cocaine or heroine use were examined in 748 homeless women. Only a third of Latinas wanted to stop using alcohol; they were also at relatively high risk for continued heroine use. Recognition that their substance use was an extremely serious problem was a consistent predictor of wanting to quit substance use. Other important predictors of desiring to quit two substances included not hanging out with other drug users, lifetime hospitalization for drug use and recent substance use treatment. Findings from this study may be helpful for selecting relative good candidates for the limited number of substance abuse treatment slots available for homeless women and providing supplementary assistance to those already in treatment. PMID- 15530727 TI - Reactivity to conspicuousness and alcohol use among college students: the moderating effect of alcohol expectancies. AB - The analysis of self-report data from 147 college undergraduates suggested that alcohol expectancies moderate the effect of a dispositional susceptibility to embarrassment elicited by undesired conspicuousness [center-of-attention-induced embarrassability (CAE)] on drinking behavior. Individuals unlikely to experience embarrassment when they engage in behaviors that make them stand out in a crowd, a common occurrence when one drinks to excess, drank heavily if they expected alcohol to make them more assertive socially. Students with similar beliefs about the effects of alcohol on social interaction who were high in CAE consumed substantially less alcohol than the latter individuals. Their overall levels of drinking were more comparable to those of the low-expectancy participants, suggesting that the disdain for conspicuousness characteristic of people with a susceptibility to CAE may counteract the desire for social disinhibition that often motivates alcohol consumption. PMID- 15530728 TI - Heterogeneity in phenotypes based on smoking status in the Great Lakes Smoker Sibling Registry. AB - We investigated whether 52 same-sex sibling pairs discordant for ever-smoking differed on psychiatric cofactors, alcohol and caffeine use, and responses to initial exposure to smoking. Ever-smokers scored significantly higher on measures of novelty seeking, depression, and childhood ADHD, and on alcohol dependence, alcohol intake, and caffeine intake. They reported significantly more pleasurable experiences, dizziness, "buzz," and relaxation upon initial exposure to smoking and significantly fewer displeasurable sensations, nausea, and cough than did nicotine-exposed, never-smoking siblings. Ever-smokers had significantly fewer years of education than their never-smoking siblings, suggesting that the concentration of smokers in lower socioeconomic strata may be partly due to downward mobility among smokers, possibly because of the observed elevation in psychiatric cofactors, which may interfere with academic performance. These findings are consistent with differences previously identified in unrelated ever- and never-smokers. Because same-sex siblings typically share a large set of common environments during childhood, our findings could be due either to genetic differences among siblings and/or (excepting educational level and responses to early exposure) to differences in adult environments. PMID- 15530729 TI - Spirituality, contentment, and stress in recovering alcoholics. AB - The relationships among length of sobriety, spirituality, contentment, and stress were examined in a sample of recovering alcoholics attending Alcoholics Anonymous meetings. Length of sobriety was significantly associated with spirituality but not with contentment or stress. Spirituality and contentment were also positively related. There were significant gender differences in the relationships among spirituality, contentment, and stress level. Spirituality was significantly correlated with lower stress for women but not for men. Men's contentment was significantly related to lower stress levels, but contentment and stress was nonsignificant for women. PMID- 15530730 TI - Differences between problem and nonproblem gamblers in subjective arousal and affective valence amongst electronic gaming machine players. AB - Arousal-based theories of gambling suggest that excitement gained from gambling reinforces further gambling behavior. However, recent theories of emotion conceptualize mood as comprising both arousal and valence dimensions. Thus, excitement comprises arousal with positive valence. We examined self-reported changes in arousal and affective valence in 27 problem and 40 nonproblem gamblers playing electronic gaming machines (EGMs). Problem gamblers reported greater arousal increases after gambling and increases in negative valence if they lost. This accords poorly with an excitement-based explanation of problem gambling. PMID- 15530731 TI - Parental prompts as risk factors for adolescent trial smoking: results of a prospective cohort study. AB - Our earlier cross-sectional research suggested that smoking parents, particularly Latino parents, engage in behaviors that may prompt their children to smoke (e.g., request their child to start the parent's cigarette in his/her own mouth). This prospective study of 478 adolescent never-smokers, mostly Latino, suggests that parental prompts to smoke were not significantly related to smoking initiation among adolescents over a 1-year period. PMID- 15530732 TI - Impact of attention-deficit hyperactivity disorder and other psychopathology on treatment retention among cocaine abusers in a therapeutic community. AB - Although there are some data suggesting that individuals with depressive disorders may be more likely to remain in treatment than those without depressive disorders, it is less clear how well other psychiatric subgroups compare to those without psychiatric comorbidity. This sample is a follow-up study of 135 individuals who were admitted into a therapeutic community. Individuals with attention-deficit hyperactivity disorder (ADHD), other Axis I disorders (no ADHD), and no Axis I disorders were compared. Although individuals with other Axis I disorders had a strikingly low early drop-out rate, after a prolonged time in treatment, the drop-out rate increased substantially, such that these individuals were found to complete treatment at a lower rate (17%) than those with no Axis I disorders (29%). Furthermore, individuals with ADHD were less likely to graduate treatment than those with other Axis I or no Axis I disorders (0%, 9%, and 19%, respectively). Future investigations may be useful to determine whether pharmacologic or nonpharmacologic interventions might improve treatment outcome. PMID- 15530733 TI - Treating addictive behaviors in the employee assistance program: implications for brief interventions. AB - Employee assistance programs (EAPs) are widely available to assist employees with a variety of problems. This research examined factors related to utilization and outcome by individuals with addictive behaviors (ABs) versus other problem areas. The specific aims of this study were to evaluate referral source and treatment outcome by gender and presenting problem. The sample included 3890 men and women who attended the EAP for a variety of concerns. Men were less likely than women to self-refer and more likely to be mandated to the EAP. Men were also much more likely to present with ABs. Relative to clients presenting with other issues, individuals with ABs were less likely to self-refer, have their problems resolved in the EAP, and were seen for fewer sessions. These results suggest that EAPs may be well suited for implementation of brief interventions (BIs) that have been empirically supported in other contexts. PMID- 15530734 TI - Utilizing virtual reality to standardize nicotine craving research: a pilot study. AB - Traditional cue reactivity provides a methodology for examining drug triggers and stimuli in laboratory and clinical settings. However, current techniques lack standardization and generalization across research settings. Improved methodologies using virtual reality (VR) cue reactivity extend previous research standardizing exposure to stimuli and exploring reactions to drug cues in a controlled VR setting. In a controlled pilot trial, 13 nicotine-dependent participants were allowed to smoke ad libitum then exposed to VR smoking and VR neutral cues and compared on craving intensity. VR smoking cues significantly increased craving compared to VR neutral cues. On average, craving intensity increased 118% during exposure to VR smoking cues. Implications for substance abuse research and treatment using VR to assess cessation and anticraving medications are discussed. PMID- 15530735 TI - Characterization of BoHV-1 gE envelope glycoprotein mimotopes obtained by phage display. AB - A phage-displayed peptide library was screened using four mAbs directed against bovine herpesvirus 1 (BoHV-1) gE glycoprotein to identify peptides mimicking this glycoprotein. The selected mimotopes allowed us to characterize the epitopes corresponding to the mAbs as continuous and proteinic and to consider using these peptides in further studies. One epitope has been clearly located at the C terminus of the protein (amino-acids 561-569). The three other mAbs enabled us to stress the immunogenic relevance of the proline-rich motifs of gE. Selected peptides showed no clear sequence identity with gE, but there is a clear link between gE proline-rich regions and the amino-acid composition of the mimotopes. The proline-rich motifs of gE are potentially located in flanking regions involved in the gE/gl glycoprotein complex formation. N-terminal fusion to pill or pVIII filamentous phage protein, C-terminal fusion to the T7 phage capsid protein, biotinylated synthetic peptides and insertion between the non-cleaved CX leader sequence and the C-terminal part of Caulobacter crescentus RsaA protein have been tested in order to increase the valency of a model peptide. We have diverted the C. crescentus expression system and proven its usefulness using the RsaA protein as a scaffold displaying the peptides of interest. Comparison between these different display systems in an indirect ELISA, indicates that the C. crescentus expression and the T7 phage display systems have some major advantages. PMID- 15530736 TI - Antigenic and genotypical characterization of Newcastle disease viruses isolated in Taiwan between 1969 and 1996. AB - Three major epidemics of Newcastle disease (ND) occurred in Taiwan over the past three decades (in 1969, 1984, and 1995). In order to gain a better understanding of the relationships between past ND epizootics in Taiwan, 36 ND viruses (NDVs) isolated between 1969 and 1996 were characterized antigenically and genotypically. The antigenicity of these viruses was analysed by their ability to cause binding of mouse monoclonal antibodies (mAbs) to cell cultures infected with the isolate. Using a panel of 22 mAbs to divide NDVs into subgroups, a total of 18 binding patterns were revealed. The sequences covering the cleavage site of the fusion protein gene of these isolates were also determined. The results of the phylogenetic analysis placed 36 NDVs into I, II, VIb, VIIa, VIII and two novel genotypes (provisionally termed X and VIh). The 1969 velogenic isolates were of genotypes X and VIh; the 1984-1985 velogenic isolates were genotyped VIb, VIh, VIIa, and X; while the 1995-1996 velogenic isolates were genotyped VIIa or VIII. Some 1969 and 1984 velogenic isolates were of the same mAbs binding pattern and genotype, and the mAbs binding patterns of the 1995-1996 isolates have not been seen before. It is concluded that velogenic NDVs of different genotype and antigenic type have co-circulated in Taiwan at least since 1969. Also there were epizootiological links between strains isolated in 1969 and 1984, whereas the 1995-1996 epidemic was caused by new antigenic variants. PMID- 15530737 TI - Proposal to reclassify Paenibacillus larvae subsp. pulvifaciens DSM 3615 (ATCC 49843) as Paenibacillus larvae subsp. larvae. Results of a comparative biochemical and genetic study. AB - The bacterial pathogen Paenibacillus larvae subsp. larvae (P. l. larvae), is the etiological agent of American foulbrood, an extremely contagious and disastrous disease of honeybee brood. In case of American foulbrood the destruction of infected colonies is often considered the only workable control measure. Therefore, the ability to diagnose this disease properly is important to prevent unnecessary economic loss to beekeepers. The development of suitable methods for the early and reliable detection of P. l. larvae is hampered by the fact that the two subspecies of Paenibacillus larvae, P. l. larvae and Paenibacillus larvae subsp. pulvifaciens (P. l. pulvifaciens), seem to be indistinguishable by cultural characteristics as well as by PCR protocols. Here we present an extensive analysis of several P. larvae reference strains. We employed conventional culture techniques, morphological and biochemical identification, PCR-based methods and sequencing of the 16S rDNA. We found indeed that P. l. pulvifaciens strain DSM 3615 is indistinguishable from P. l. larvae (DSM 7030). We did not face any problems to discriminate between P. l. larvae and P. l. pulvifaciens strains DSM 8442 and DSM 8443. Therefore, classification of DSM 3615 as type strain of P. l. pulvifaciens seems not to be justified. We propose to reclassify this strain as P. l. larvae. Former problems in differentiating the two subspecies might have arisen from this misclassification. PCR-based methods as well as appropriate biochemical identification systems provide a reliable means for the discrimination between the two subspecies P. l. larvae and P. l. pulvifaciens. PMID- 15530738 TI - Longitudinal study of interferon-gamma, serum antibody and milk antibody responses in cattle infected with Mycobacterium avium subsp. paratuberculosis. AB - During a 2-year study period, 252 animals from dairy herds infected with Mycobacterium avium subsp. paratuberculosis, and 119 animals from non-infected herds were subjected to repeated blood and faecal sampling. Animals were retrospectively grouped by infection status as infected, exposed (culture negative animals from infected herds), or non-infected animals, and by age, 12-23 months (1+ year), 24-35 months (2+ years), or 36 months and older (3+ years). Samples were collected for culture of faeces, assessment of interferon-gamma (IFN gamma) secreted by M. paratuberculosis antigen stimulated whole-blood lymphocytes (IFN-gamma test), and measurement of antibody responses against M. paratuberculosis in serum and milk by an in-house absorbed ELISA. The IFN-gamma test diagnosed higher proportions of infected and exposed animals than the antibody ELISAs. The highest sensitivity of IFN-gamma test was in infected cattle of 2+ years of age. Receiver-operating characteristic (ROC) analyses supported the assumption that the IFN-gamma test had a better performance than antibody tests of animals of 1+ and 2+ years of age. However, for animals of 3+ years all tests performed equally well. Application of single sampling compared with repeated samplings showed better performance of the IFN-gamma test by repeated samplings, and the milk antibody ELISA in animals of 3+ years of age performed significantly better with repeated sampling compared with single sampling. In conclusion, the IFN-gamma test may be applied for screening of cattle of 1 and 2 years of age for exposure to M. paratuberculosis and the antibody ELISAs from 3 years of age. PMID- 15530739 TI - Invasion of chicken embryo fibroblast cells by avian Pasteurella multocida. AB - Invasion of chicken embryo fibroblast (CEF) cells by the virulent encapsulated Pasteurella multocida strains P-1059 (serovar A:3) and X-73 (serovar A:1) and an avirulent noncapsulated derivative P-1059B (serovar -:3) was investigated. The number of intracellular bacteria increased for all the strains after 2, 4 and 6 h post-inoculation to CEF cells. By 6 h post-inoculation, the number of invaded bacteria of encapsulated strains was significantly higher than noncapsulated strain and reached 150- and 112-fold for strains P-1059 and X-73, respectively, while it was 9-fold for strain P-1059B as compared to the number of invaded bacteria recovered after 2 h post-inoculation. Electron microscopy of invasion by encapsulated strains showed that the bacteria were adhering to CEF cells membrane after 1 h of inoculation. By 4-h, one or two bacteria were detected within membrane-bound vacuoles of the intracellular space. The number of intracellular bacteria markedly increased at 14 h post-inoculation. Invasion of all strains was inhibited significantly when the monolayers were treated with periodic acid (P<0.001) or trypsin (P<0.05). The treatment of bacteria with hyaluronidase did not affect invasion. The present results indicate that avian P. multocida capsular type A strains are invasive and that the receptor on CEF cell surface might be glycoprotein. PMID- 15530740 TI - Risk assessment of transmission of capsule-deficient, recombinant Actinobacillus pleuropneumoniae. AB - Actinobacillus pleuropneumoniae is the etiologic agent of swine pleuropneumonia. Live, non-encapsulated vaccine strains have been shown to be efficacious in preventing acute disease in pigs. Recombinant DNA technology has the advantage of generating defined mutants that are safe, but maintain critical immunoprotective components. However, some recombinant strains have the disadvantage of containing antibiotic resistance genes that could be transferred to the animal's normal bacterial flora. Using DNA allelic exchange we have constructed attenuated, capsule-deficient mutants of A. pleuropneumoniae that contain a kanamycin resistance (Kn(R)) gene within the capsule locus of the genome. Following intranasal or intratracheal challenge of pigs the encapsulated parent strains colonized the challenge pigs, and were transmitted to contact pigs. In contrast, the capsule-deficient mutants were recovered only from the challenged pigs and not from contact pigs. Each kanamycin-resistant colony type recovered from the respiratory or gastrointestinal tracts of pigs challenged with the recombinant strain was screened with a probe specific for the Kn(R) gene. All probe-positive colonies were assayed for the specific Kn(R) gene by amplification of a 0.9 kb fragment of the antibiotic resistance gene by PCR. The 0.9 kb fragment was amplified from the recombinant A. pleuropneumoniae colonies, but not from any of the heterologous bacteria, indicating there was no evidence of transmission of the Kn(R) gene to resident bacteria. Following aerosol exposure of 276 pigs with recombinant, non-encapsulated A. pleuropneumoniae the recombinant bacteria were not recovered from any nasal swabs of 75 pigs tested or environmental samples 18 h after challenge. Statistical risk analysis, based on the number of kanamycin resistant colonies screened, indicated that undetected transmission of the Kn(R) gene could still have occurred in at most 1.36% of kanamycin-resistant bacteria in contact with recombinant A. pleuropneumoniae. However, the overall risk of transmission to any resident bacteria was far lower. Our results indicate there was little risk of transmission of capsule-deficient, recombinant A. pleuropneumoniae or its Kn(R) gene to contact pigs or to the resident microflora. PMID- 15530741 TI - Foal IgG and opsonizing anti-Rhodococcus equi antibodies after immunization of pregnant mares with a protective VapA candidate vaccine. AB - The aim of this study was to evaluate serum IgG antibody levels and opsonizing activity in foals from pregnant mares immunized with either proteins from an R. equi strain containing virulence-associated protein A (VapA), an immunodominant surface-expressed lipoprotein encoded by a virulence plasmid crucial for virulence in foals, or a whole killed virulent R. equi preparation. Forty-eight pregnant mares were distributed into three groups, i.e. 24 immunized with R. equi VapA protein antigen associated with a water-based nanoparticle adjuvant (Montanide IMS 3012), 8 immunized with whole killed R. equi, and 16 non-immunized as control. Serum IgG and opsonizing capacity were evaluated during pregnancy in mares, and up to day 45 post-delivery in foals in which R. equi infections were recorded in the first 6 months of life. Pregnant mares immunized with virulent R. equi proteins developed higher serum IgG and opsonic activity which were transferred to the foals than either in the whole R. equi immunized or the control group. Four foals developed pneumonia in the control group while none in immunized groups. Results support further evaluation of VapA protein antigen associated with a water-based nanoparticle adjuvant as a candidate vaccine for immunization of pregnant mares resulting in passive antibody-mediated protection of foals. PMID- 15530742 TI - A Lawsonia intracellularis transmission study using a pure culture inoculated seeder-pig sentinel model. AB - Transmission of Lawsonia intracellularis from experimentally inoculated pigs to naive swine was demonstrated in this study. The study was conducted using conventional pigs divided into three groups as follows: principles inoculated with L. intracellularis, sentinels, and controls. The pigs were inoculated and paired on 13 and 9 days post-inoculation with a sentinel pig for 7 days. Fecal samples and serum samples were collected throughout the study for polymerase chain reaction (PCR) and antibody testing by indirect fluorescent antibody techniques. After co-mingling, the inoculated group was necropsied; sentinel and control pigs were necropsied 7-14 days later. The intestinal tracts were evaluated grossly and microscopically for lesions. PCR was performed on intestinal mucosal scrapings and feces. Warthin-Starry and fluorescent antibody staining procedures were conducted to confirm colonization with L. intracellularis. Gross and microscopic lesions typical of porcine proliferative enteropathy (PPE) were observed in both the inoculated and sentinel groups. Transmission was demonstrated from inoculated principle pigs to sentinel pigs. PCR results detected cyclical shedding of L. intracellularis in the feces. Seroconversion occurred in pigs that were exposed to L. intracellularis. From this study, it was demonstrated that transmission of L. intracellularis can occur easily in an environment with experimentally infected pigs and that PCR can be a useful tool to monitor fecal shedding of the organism. PMID- 15530743 TI - Molecular epidemiology of avian pathogenic Escherichia coli (APEC) isolated from colisepticemia in poultry. AB - The molecular biology and epidemiology of 150 avian pathogenic Escherichia coli strains (APEC) isolated from septicemic poultry in Germany was investigated by serotyping, pulsed field gel electrophoresis (PFGE), and polymerase chain reaction (PCR). Only 49.6% of the isolates could be grouped to serogroups O1, O2, and O78. Macrorestriction analyses data revealed two large clonal groups (clusters I and II) among the APEC strains with a similarity of 60.9% to each other. An association between restriction pattern and serogroup or origin of the strains was only present in a few subgroups of each clusters I and II, but was not evident. In contrast, our data revealed distinct combinations of virulence associated genes in that 51.2% of the O2-strains harboured a combination of the genes fyuA, irp2, iucD, tsh, vat, fimC, and colV and 36.4% of the O78-strains possessed the same gene combination with exception of vat. With 34 different gene combinations the non-O1, -O2, -O78 isolates revealed a higher variability in their virulence gene pattern than O1-, O2-, and O78-strains with 6, 13, and 9 patterns, respectively. Our data indicate only a limited association between the virulence gene pattern and the serogroup of APEC strains and question the sensitivity of O-typing for APEC identification without the application of further diagnostic tools. Although a limited number of APEC clones exist, horizontal gene transfer seems to be common in these pathogens. These findings strengthen further research on the population structure of APEC and may be the reason for the lack of clear definition of this common E. coli pathotype. PMID- 15530744 TI - Physiological and genetic characterisation of some new Aphanomyces strains isolated from freshwater crayfish. AB - Five Aphanomyces strains were isolated during suspected outbreaks of crayfish disease in Spain and Italy. Genetic and physiological evidence show that the strains isolated from the freshwater crayfish Procambarus clarkii and Pacifastacus leniusculus, do not fit into any previously identified group of Aphanomyces astaci and are not capable of killing crayfish following standardised experimental infection. RAPD-PCR and ITS sequencing analysis show a high degree of similarity between the new isolates, while they are clearly different from the A. astaci reference strains. They do, however, possess some properties, which are commonly associated with parasitic species such as repeated zoospore emergence and the lack of sexual reproduction. The five isolates share some physiological properties i.e. a high growth rate, and germination in response to nutrients and, in contrast to A. astaci, they do not express chitinase constitutively during growth or sporulation. Until their taxonomic status is fully elucidated we suggest that the new isolates be given the tentative species name Aphanomyces repetans. PMID- 15530745 TI - Molecular characterization of porcine TT virus, an orphan virus, in pigs from six different countries. AB - Human TT virus (TTV), originally isolated from a patient with post-transfusion hepatitis in 1997, is ubiquitous and non-pathogenic. Viruses related to human TTV have since been identified in non-human primates, bovine, ovine, porcine, feline, and canine. The objective of this study was to genetically characterize porcine TTV from pigs in different geographic regions. PCR primers based on the non coding region of the only available porcine TTV isolate were designed to amplify porcine TTV DNA from sera of pigs in six different countries. Porcine TTV DNA was detected in 66.2% (102/154) of the swine sera. The percentages of positive pigs varied greatly from country to country and even within the same country: 33% in Iowa, USA; 40% in Thailand; 46% in Ontario, Canada; 80% in China; 85% in Korea; 90% in Spain; 100% in Quebec and Saskatchewan, Canada. A total of 40 porcine TTV isolates (five from each geographic region) were sequenced for a 218 bp fragment within the non-coding region. Sequence analyses revealed that porcine TTV isolates from different geographic regions shared 86-100% nucleotide sequence identity to each other. The prototype Japanese isolate of porcine TTV, Sd-TTV31, shared 90-97% nucleotide sequence identity with porcine TTV isolates reported in this study. Phylogenetic analysis showed that the clustering of the porcine TTV isolates is not associated with geographic origins. Although porcine TTV is not known to be associated with any swine disease, co-infection of pigs with TTV and other known swine pathogens may result in enhanced disease. There are also concerns for risk of potential human infection during xenotransplantation. PMID- 15530746 TI - Interaction between attaching and effacing Escherichia coli serotypes O157:H7 and O26:K60 in cell culture. AB - Ruminants harbour both O157:H7 and non-O157 Attaching Effacing Escherichia coli (AEEC) strains but to date only non-O157 AEEC have been shown to induce attaching effacing lesions in naturally infected animals. However, O157 may induce lesions in deliberate oral inoculation studies and persistence is considered dependent upon the bacterially encoded locus for enterocyte effacement. In concurrent infections in ruminants it is unclear whether non-O157 AEEC contribute either positively or negatively to the persistence of E. coli O157:H7. To investigate this, and prior to animal studies, E. coli O157:H7 NCTC 12900, a non-toxigenic strain that persists in conventionally reared sheep, and non-toxigenic AEEC O26:K60 isolates of sheep origin were tested for adherence to HEp-2 tissue culture alone and in competition one with another. Applied together, both strains adhered in similar numbers but lower than when either was applied separately. Pre incubation of tissue culture with either one strain reduced significantly (P < 0.05) the extent of adherence of the strain that was applied second. It was particularly noticeable that AEEC O26 when applied first reduced adherence and inhibited microcolony formation, as demonstrated by confocal microscopy, of E. coli O157:H7. The possibility that prior colonisation of a ruminant by non-O157 AEEC such as O26 may antagonise O157 colonisation and persistence in ruminants is discussed. PMID- 15530747 TI - A specific PCR for the identification of Mycoplasma capricolum subsp. capripneumoniae, the causative agent of contagious caprine pleuropneumonia (CCPP). AB - Contagious caprine pleuropneumonia is a severe infectious disease of goats in Africa and the Middle East. It is caused by a fastidious mycoplasma, Mycoplasma capricolum subsp. capripneumoniae, a member of the "M. mycoides cluster". Members of this cluster share genomic and antigenic features, which result in common biochemical and serological properties, complicating species identification. Two species of this cluster, M. mycoides subsp. capri and M. mycoides subsp. mycoides large colony biotype, are very often isolated from clinical cases resembling contagious caprine pleuropneumonia. Furthermore, in the laboratory, M. capricolum subsp. capripneumoniae can be easily confused with the closely related capricolum subspecies. Considering these constraints and the scarcity of available methods for identification, a specific polymerase chain reaction was developed. A DNA fragment of 7109 bp containing genes coding for the arginine deiminase pathway (ADI) was chosen as target sequence for the selection of a specific primer pair. The full ADI operon from M. capricolum subsp. capripneumoniae strain GL100 was sequenced. Polymorphism within this locus was analyzed by comparison with the sequence from the closely related IPX strain (M. capricolum subsp. capricolum). It varied from 0.6% to 3.5%. The highest divergence was found in a region coding for arcD. Therefore, this gene was chosen as target for the specific amplification of a 316 bp-long DNA fragment. The specificity of this PCR was validated on 14 M. capricolum subsp. capripneumoniae strains and 27 heterologous strains belonging to the "M. mycoides cluster" and M. putrefaciens. This new PCR will be a valuable tool for the surveillance of contagious caprine pleuropneumonia. PMID- 15530748 TI - High prevalence of multiple resistance to antibiotics in Salmonella serovars isolated from a poultry slaughterhouse in Spain. AB - Salmonellosis is a major foodborne infection in Spain, and strains that are resistant to a great variety of antibiotics have become a major public health concern. The aim of this study was to determine the level of antibiotic resistance in 133 Salmonella isolates obtained from a poultry slaughterhouse in Zaragoza (NE Spain). Antimicrobial resistance testing was performed by disk diffusion method using 19 antibiotics. Results were interpreted following the NCCLS criteria. Overall, the highest percentage of resistance was found to the following antimicrobial agents: sulfadiazine (96.2%), neomycin (53.4%), tetracycline (21.8%), and streptomycin (11.3%). All isolates were found to be resistant to one or more of the antibiotics tested. Multiple resistance was observed in 87 strains (65.4%). We found 23 different patterns of resistance in Salmonella Enteritidis. Resistance to sulfadiazine was the most common single resistance. The most frequent patterns of multiresistant strains were neomycin+sulfadiazine and neomycin+tetracycline+sulfadiazine. S. 4,5,12:b: showed the highest percentages of resistance to the tested drugs, with five different resistance patterns found. Ampicillin+chloramphenicol+streptomycin+sulphonamides+tetracycline (ACSSuT) resistance pattern, commonly associated with S. Typhimurium DT 104, was not detected in strains of the same phage type from broilers. The appearance of substantial multiresistance in foodborne Salmonella isolates suggests the need for more prudent use of antibiotics by farmers, veterinarians, and physicians. PMID- 15530749 TI - The evaluation of self management of health problems and the value of randomized clinical trials. PMID- 15530750 TI - The short-term effects of a body awareness program: better self-management of health problems for individuals with chronic a-specific psychosomatic symptoms. AB - A three-day residential Body Awareness Program (BAP) was developed to teach people with Chronic A-specific Psychosomatic Symptoms (CAPS) to react adequately to disturbances of the balance between a daily workload and the capacity to deal with it. The short-term effects of the program for people with CAPS are presented in this study. The design is a non-control group design with pre- and post measures (2 months after the program). The sample for this paper was formed by 187 participants. The mean age is 42.3 (S.D. = 8.9), and 57% is female. The results showed decreased stress-related symptoms, increased quality of life, increased self-efficacy, less depressive attribution style, more expression of emotions, and a positive change of lifestyle. Most of these measured changes can be interpreted as clinically relevant outcomes with medium-to-large effect sizes. Personal pre-training goals were attained by 85% of the participants. Spouses also confirmed the found effects. Evaluation of the BAP gives enough evidence to conclude that this program leads to positive effects in CAPS. Participants react more adequately to disturbances between daily workload and the capacity to deal with this load. They are more capable of self-management in coping with stress and psychosomatic symptoms. Conclusions are drawn about the prevention by early interventions for patients with a-specific physical symptoms. PMID- 15530751 TI - Self-care issues from the perspective of individuals with chronic obstructive pulmonary disease. AB - This study explored factors that influence self-care from the perspective of individuals with Chronic Obstructive Pulmonary Disease (COPD). Seven focus groups were held with individuals who had physician diagnosed COPD and experienced daily symptoms that limited activities. Forty-two subjects participated in the study. All sessions were audiotaped, transcribed and coded independently. The main theme identified was surviving COPD: the context for living and the two sub-themes identified were adjusting physically and emotionally to COPD. Participants discussed their self-care practices within the context of the purpose and meaning of life. Although participants experienced major physical limitations, the issues highlighted were those of individuals striving to survive with COPD and to hold on to some quality of life through adjusting physically and emotionally. Individuals with COPD have developed strategies to adapt to the disease, compensate for limitations, and integrate self-care activities for managing COPD into their daily routine. Health professionals should use approaches that support the whole needs of the individual to achieve the best quality of life for individuals with COPD and their families. PMID- 15530752 TI - A qualitative evaluation of a comprehensive self-management programme for COPD patients: effectiveness from the patients' perspective. AB - The COPE self-management programme, including a self-management education course, self-treatment of exacerbations and a fitness programme, appeared to have no significant effect on health related quality of life (HRQoL) as measured by the St. George's Respiratory Questionnaire (SGRQ). This is in contrast to our hypothesis and despite expressions of satisfaction of patients to healthcare workers. To understand this discrepancy, a qualitative study was performed. A purposive sample of 20 participants of the COPE self-management programme were interviewed at home using in-depth, semi-structured interviews. Interviews were audio taped and transcribed verbatim and analysed according grounded theory. The fitness programme was most positively evaluated by patients due to the perceived increase of exercise capacity and the social aspect of the group training. Major effects gained by the self-management education course reported by patients were the skills to evenly distribute their energy and to listen to their body signals. Most patients thought favourable about self-treatment of exacerbations. The possibility to start early, not having to call a doctor and autonomy were raised as important advantages. Furthermore, several patients reported increased self confidence and coping behaviour as important effects of the COPE programme. Finally, many patients reported feeling safe due to the frequent follow-up visits and 24h access to the hospital, and this aspect elicited to be very important. In this study, the qualitative interviews suggest that the SGRQ and possibly other existing HRQoL instruments might fail to capture the full experience of patients in self-management studies. The need for more elaborate qualitative research on this subject is indicated. PMID- 15530753 TI - Patients evaluations of their health care: the expression of negative evaluation and the role of adaptive strategies. AB - Patient experiences surveys have become common within healthcare and represent an important form of participation, with patients having the potential ability to influence the quality of care. However, there is still a relatively limited understanding of how patients evaluate the quality of their care. We present the findings of a qualitative study, which explored the way in which patients express their evaluations, in an attempt to move the debate forward. The results indicate that patient evaluation appears to be far more complex than routinely acknowledged, particularly in relation to the expression of negative experiences. Patients were reluctant to offer directly negative criticisms, needed particular conditions in which to express their negative evaluations and used a variety of adaptive strategies to overcome social pressures that inhibited negative evaluation and promoted positive evaluation. We argue that standardised approaches to measuring negative evaluations, such as questionnaires, need to become more sensitive to the complexities of negative evaluation. This would enhance the extent to which patients can participate in influencing the quality of care. A number of recommendations are made to develop the sensitivity of future investigations into the negative aspects of patient experiences. PMID- 15530754 TI - The effectiveness of physical activity counseling in a work-site setting. A randomized, controlled trial. AB - Voluntary employees (N = 155) from nine different companies were screened by questionnaire for the study. They were randomized into three study groups: counseling (n = 52), counseling + fitness testing (n = 51) and control group (n = 52). The counseling was based on a goal-oriented conversation session for each participant and three follow-up appointments with an occupational nurse over a period of 1 year. The fitness tests were adapted from the UKK Health-related Fitness Test Battery. The outcome measures were the changes in the amount of leisure-time physical activity (LTPA) assessed by diary, pedometer and questionnaire at baseline and at 6 and 12 month follow-up visits. As a result, no statistically significant differences were detected between the three groups at either of the follow-up visits. It seemed, thus, that the two PA counseling methods implemented had no direct mid- or long-term effects on the LTPA of voluntary employees with no specific disease-related indication to increase LTPA. PMID- 15530755 TI - A training and support programme for caregivers of children with disabilities: an exploratory study. AB - Our aim was to conduct an exploratory evaluation of an intervention designed to equip caregivers with simple massage skills that they could use with their children in the home. The sample comprised 82 parents and 82 children with disabilities. Data were collected by self-administered questionnaires at two points in time: baseline, and immediately post-programme (after 8 weeks). Qualitative data were collected through Home Record Sheets and Monitoring Forms completed by caregivers and therapists, respectively. Results revealed statistically significant improvements in caregivers' self-efficacy in their ability to conduct massage, in managing children's psychosocial wellbeing and in levels of anxious mood. Caregiver reports of children's sleeping patterns and eating showed significant improvements. Qualitative data confirmed that caregivers believed that children's sleep patterns improved and also suggested improvements in children's bowel movements, awareness of their bodies, movement, and communication; factors that could be included in future evaluation. PMID- 15530756 TI - Improving client-provider communication: evaluation of a training program for women, infants and children (WIC) professionals in New York state. AB - Results are presented from evaluation of an intensive 1 day training program to improve the growth monitoring counseling skills of Special Supplemental Nutrition Program for Women, Infants and Children (WIC) providers. The training was framed by the patient-centered approach, and focused on a seven-step technique that emphasized eliciting client perspective on the child's health and negotiating follow-up strategies. Changes in skill were assessed during audiotaped mock counseling sessions with simulated clients. Observed intervention effects were moderate but encouraging for future training programs. After the training, more providers elicited client perspective, and provider level of engagement in negotiating with the client increased. At post-test providers asked more open ended questions than at pre-test, and provider-to-client talk ratio decreased. Increases in provider total and competence-related satisfaction paralleled improvements in counseling proficiency. Study results suggest that counseling skills of non-physician health providers can change after a 1 day focused training: providers were more client-centered in their discussions. Limitations and implications of the study are discussed. PMID- 15530757 TI - Increasing screening uptake amongst those intending to be screened: the use of action plans. AB - This experimental study investigates an intervention designed to increase rates of antenatal screening uptake in those intending to undergo antenatal screening. Eighty-eight pregnant women intending to undergo prenatal screening were alternately allocated to standard care or asked to write a simple plan for attending or making an appointment. Twenty-five (63%) in the intervention group made an action plan. There was no difference in uptake of screening between the intervention group and the control group, nor between those making an action plan and the control group. Within the intervention group, those making an action plan had higher screening uptake (21/25, 84%) than those not making one (7/15, 47%; CI(95) difference = 8-66%, P = 0.017). Asking women who intend to undergo screening to make an action plan does not increase screening uptake. This result raises doubts about introducing simple action plans in a clinical situation as an effective means of changing behaviour. PMID- 15530758 TI - Exhaustion and coronary heart disease: the history of a scientific quest. AB - This paper describes the development of the concept "vital exhaustion", the epidemiological evidence that this state belongs to the precursors of different manifestations of coronary artery disease, and the biological plausibility of the association between exhaustion and coronary artery disease. It further describes the main results of a randomized, controlled trial designed to determine whether a behavioural intervention reduces the risk of a new coronary event. PMID- 15530759 TI - Balint training makes GPs thrive better in their job. AB - In this study, we examined Balint group participants' sense of control and satisfaction in their work situation and their attitudes towards caring for patients with psychosomatic problems. Forty-one GPs filled in a questionnaire with a 10-point visual analogue scale. Of these, 20 had participated in Balint groups for more than one year and 21 were a reference group. The Balint physicians reported better control of their work situation (e.g. taking coffee breaks and participating in decision making), thought less often that the patient should not have come for consultation or that psychosomatic patients were a time consuming burden, and were less inclined to refer patients or take unneeded tests to terminate the consultation with the patient. These results might indicate higher work-related satisfaction and better doctor-patient relationship. PMID- 15530760 TI - The influence of locus of control on preferences for information and decision making. AB - Previous studies reported inconsistent relationship between the preference for information and that for decisional autonomy in medical encounters. We hypothesized that the relationship may be dependent on people's attribution beliefs on health. To test the hypothesis, demographically representative sample of Japanese population over the age of 18 (N = 3395) were invited into self administered questionnaire survey. Preferences for information and decisional autonomy were measured by a translated version of autonomy preference index. Health attribution was measured by a scale modified from multi-dimensional health locus of control scales. After adjusting for socio-economic status, multivariable linear regression analysis revealed that information preference was positively associated with decisional preference among individuals who believed their health is less dependent on influential others. However, the reversed relationship was observed in case of individuals with attribution to others. The results suggested that individuals may use medical information for different purposes according to the types of health-related attribution beliefs. PMID- 15530761 TI - Building concordant relationships with patients starting antidepressant medication. AB - Fifty one patients beginning courses of antidepressant medication were recruited from general practice surgeries in two health authorities in London and interviewed on two occasions. Respondents were asked about their experiences of using antidepressant medication, including their information needs and the level of involvement which they had had in making decisions about their medication. Analysis was carried out using qualitative procedures. The study identified information needs which were unmet at the start of treatment, including the impact of adverse drug reactions on patients' lives, the very gradual process of recovery and information regarding dosage prescribed in the context of minimum and maximum dosages. Respondents also had a broad view of the term dependency that needed to be addressed. However, some respondents reported that they experienced difficulty in absorbing information given during the initial consultation. Therefore developing strategies for reinforcement of information would be advantageous. Patients involvement in decision making varied between respondents and at different points in therapy. PMID- 15530762 TI - Anxiety levels in women attending a colposcopy clinic: a randomised trial of an educational intervention using video colposcopy. AB - A video colposcopy allows the real-time images viewed by the doctor performing the examination to be viewed by patients on a television monitor during the consultation. Eighty-one women (requiring either laser treatment or a normal recheck) were randomly assigned to either the video intervention group or the control group. A significant decrease in state anxiety was observed from one visit to the next in all patients (P = 0.000). This decrease in anxiety was significantly greater in the laser patients in the video colposcopy condition than patients in the control group for both groups of patients. Patients in the experimental groups also reported less pain than patients in the control conditions (P < 0.05). This benefit associated with video colposcopy was not observed on the second (treatment) visit. In conclusion, video colposcopy is a useful and time-efficient method that reduces patient anxiety and pain during examination visits, but not necessarily during visits where laser treatment is required. PMID- 15530763 TI - Clinical practice guidelines to improve shared decision-making about assistive device use in home care: a pilot intervention study. AB - This study examines whether clinical practice guidelines (CPG) for the introduction of assistive devices (ADs) in home care improve shared decision making about AD use and modify its social-cognitive correlates. Data were collected in an intervention study with quasi-experimental design. Questionnaires were obtained from 116 home nurses and home care workers and their 140 clients with disabilities. Significant differences between intervention and control group revealed that implementation of CPG improved home nurses' and home care workers' self-reported practice: the number of intervention methods they applied increased, and the methods were applied with increased intensity. Nurses' attitudes towards introducing ADs in a shared decision-making process decreased in both intervention group and control group over time. Clients' reports about caregivers' practice showed a minor concurrence with the reports of the caregivers themselves. The complexity of the CPG and a substantial time investment were reported as the main barriers to involving clients in shared decision-making through the experimental CPG. Nevertheless, CPG hold promise for shared decision-making between formal caregivers and community-dwelling persons with disabilities concerning AD use. The findings suggest that these CPG can structure communication between caregiver and client, and can support caregivers in facilitating clients' self-determination concerning coping with their disabilities. PMID- 15530764 TI - Predictive testing for hereditary non-polyposis colorectal cancer: motivation, illness representations and short-term psychological impact. AB - This paper describes the motivation, recall of cancer risks, and illness representations of 40 individuals who had a predictive test for hereditary non polyposis colorectal cancer (HNPCC) as well as the short-term impact of predictive testing by means of a semi-structured interview and self-report questionnaires. The main motives for predictive testing were early detection of cancer, knowledge of the children's risk and reduction of uncertainty. Overall, recall of cancer risks was good. Measurements of illness representations revealed low perceptions of "threat" of cancer and high confidence in the controllability of the disease. Distress was within normal ranges. Distress decreased significantly from pre- to post-test in non-carriers and did not in carriers. It also decreased in individuals for whom "reducing uncertainty" was a very important motive for the test, not in the others. Although part of the carriers did not have colonoscopies, all carriers intended to have regular colonoscopies in the future. PMID- 15530765 TI - Suitability of prostate cancer education materials: applying a standardized assessment tool to currently available materials. AB - Written educational materials serve as important teaching instruments for prostate cancer patients and their families. However, they must be understandable for their intended audiences. We examined prostate cancer brochures and pamphlets using the Suitability Assessment of Materials (SAM) instrument for the materials' overall suitability, readability, and cultural appropriateness. These factors are crucial to ensure that patients are able to understand and relate to the information. Our evaluation of 29 materials demonstrated that the majority, 22 (75.8%), scored "adequate" for their overall suitability. However, 26 materials (90%) scored "not suitable" for their reading grade level, while 55% of the materials could not be rated on cultural appropriateness because of lack of cues about the intended audience. Also, many of the materials scored poorly on content, graphics, self-efficacy, and learning motivation and stimulation. Most of the materials did well with typography and layout. Overall, the findings point to the need to carefully assess written materials used for multicultural audiences with low reading ability. PMID- 15530766 TI - Patient accounts for noncompliance with diabetes self-care regimens and physician compliance-gaining response. AB - The purpose of this study was to identify diabetics' accounts for medical noncompliance, physicians' compliance-gaining strategies elicited in response to those accounts, and to examine the relationship between accounts and compliance gaining strategies. Telephone surveys of diabetics from Kentucky and Tennessee assessed patient perceptions of physician-patient communication during their last diabetes treatment-related appointment. Eighty-four patients were able to provide categorical data for analysis. Patient account strategies for noncompliance were coded as concessions, excuses, justifications, and refusals. Physician compliance gaining strategies were coded as positive-regard, negative-regard, or neutral regard strategies. Results showed the most common patient account was concession (33%), then excuse (23%), justification (22%), and refusal (7%). The most common physician compliance-gaining strategy was neutral regard (54%), then positive regard (31%) and negative regard (15%). Results also indicated that patient account and physician compliance-gaining strategy were related. Specifically, concession accounts from patients elicited a subsequent neutral-regard compliance gaining strategies from the physician in 76% of concession cases. Results also show that justifications were most likely to elicit neutral- (52%) or positive regard (43%) strategies. Implications for physician practice and future research are discussed. PMID- 15530767 TI - Literacy demands of product information intended to supplement television direct to-consumer prescription drug advertisements. AB - The US Food and Drug Administration (FDA) allows television direct-to-consumer (DTC) prescription drug advertisements that do not fully disclose drug risks if the ads include "adequate provision" for dissemination of the drug's approved labeling. This requirement can be met in part by referring consumers to multiple text sources of product labeling. This study was designed to assess the materials to which consumers were referred in 23 DTC television advertisements. SMOG assessments showed that the average reading grade levels were in the high school range for the main body sections of the materials and college-level range for the brief summary sections. The Suitability Assessment of Materials (SAM) instrument identified specific difficulties with the materials, including content, graphics, layout, and typography features. Stronger plain language requirements are recommended. Health care providers should be aware that patients who ask about an advertised drug might not have the full information required to make an informed decision. PMID- 15530768 TI - Patients' ideas, fears and expectations of their coronary risk: barriers for primary prevention. AB - The application of cardiovascular guidelines and risk tables may be impeded by many barriers. In the present paper, we explored the role of patients in the feasibility of cardiovascular preventive care in general practice. Patient related barriers were examined by means of a qualitative study. Fifteen GPs audio taped one or two consultations on primary cardiovascular preventive care. The tapes were used to guide the subsequent semi-structured in-depth interviews with patients. Twenty-two patients were interviewed. Patients' understanding of prevention of cardiovascular disease (CVD) was often insufficient. The risk table and the multi-factorial approach were difficult to understand. Risk perception was often unrealistic and dichotomous, and mainly based on personal experiences. There was a demand for more information and cholesterol tests. At the patient level, many barriers impede effective prevention of cardiovascular diseases. In particular, the highly individualized high-risk approach needs to be explained to patients. Educational patient materials, intended to support both the GP and the patient, should take into account the ideas, fears and expectations of patients. PMID- 15530770 TI - Cell cycle goes global. AB - The cell division cycle is one of the most intensively studied biological processes, yet, in spite of great effort, many questions remain as to how the cell cycle is controlled by cyclin-dependent kinases and other critical regulators. Recent functional genomic and proteomic approaches have yielded new insights into almost all aspects of cell cycle control, including transcriptional circuits, DNA replication, sister chromatid separation and regulation by environmental signals. Perhaps most notably, systematic analysis has begin to reveal meta-level connections between previously distinct sub-processes. As the interconnections between these huge datasets are beyond intuition, mathematical representation and automated analysis of functional genomic data is an urgent mandate. PMID- 15530771 TI - New tricks for old dogs: unexpected roles for cell cycle regulators revealed using animal models. AB - Studies in animal models have revealed many surprises regarding the importance of key cell cycle regulators during animal development and homeostasis, underscoring the plasticity and redundancy of cell cycle circuitry within a whole-animal context. Moreover, checkpoint regulators, which are not essential for viability in yeast and cultured cells, play important roles in cell cycle control during development. PMID- 15530772 TI - Cell cycle, proteolysis and cancer. AB - Research in the past 15 years has shown that the mammalian cell cycle is controlled by the action of cyclin-dependent kinases (CDKs). A crucial substrate of the CDKs in G1-phase is the retinoblastoma tumor suppressor (pRB), which restrains proliferation largely by repressing the activity of the E2F transcription factors. More recent work has shown that the cell cycle is also a tale of two classes of ubiquitin ligases, referred to as SCF and APC/C ligases. CDKs, E2F and ubiquitin ligases reciprocally regulate each other, resulting in complex feedback loops. Perturbation of this network of molecular machines is associated with proliferative diseases, including cancer. PMID- 15530773 TI - The DNA damage response: sensing and signaling. AB - The protein kinases ATM and ATR are central components of the checkpoint mechanisms that signal the presence of damaged DNA and stalled replication forks. Recent studies have provided important new insights into how these kinases work together with their regulatory subunits, DNA repair proteins and adaptor proteins to sense abnormal DNA structures and implement the appropriate DNA damage response. These advances have provided a more detailed understanding of the interface between damaged DNA and the checkpoint sensor proteins. PMID- 15530774 TI - Coordination of cell death and the cell cycle: linking proliferation to death through private and communal couplers. AB - In development and in the adult, complex signaling pathways operate within and between cells to coordinate proliferation and cell death. These networks can be viewed as coupling devices that link engines driving the cell cycle and the initiation of apoptosis. We propose three simple frameworks for modeling the effects of proliferative drive on apoptotic propensity. This perspective offers a potentially useful foundation for predicting group behaviors of cells in normal and pathological settings. PMID- 15530775 TI - Life's smile, death's grin: vital functions of apoptosis-executing proteins. AB - Apoptosis is executed by caspases as well as caspase-independent death effectors. Caspases are expressed as inactive zymogens in virtually all animal cells and are activated in cells destined to undergo apoptosis. However, there are many examples where caspase activation is actually required for cellular processes not related to cell death, namely terminal differentiation, activation, proliferation, and cytoprotection. Several caspase-independent death effectors including apoptosis-inducing factor, endonuclease G and a serine protease (Omi/HtrA2) are released from the mitochondrial intermembrane space upon permeabilization of the outer membrane. Such proteins also have important roles in cellular redox metabolism and/or mitochondrial biogenesis. As a general rule, it thus appears that cell-death-relevant proteins, especially those involved in the core of the executing machinery, have a dual function in life and death. This has important implications for pathophysiology. The fact that the building blocks of the apoptotic machinery have normal functions not related to cell death may mean that essential parts of the apoptotic executioner cannot be lost and thus reduces the possibility of oncogenic mutations that block the apoptotic program. Moreover, therapeutic suppression of unwarranted cell death must be designed to target only the lethal (and not the vital) role of death effectors. PMID- 15530776 TI - Regulation of mitochondrial membrane permeabilization by BCL-2 family proteins and caspases. AB - Mitochondria play an important role in the integration and transmission of cell death signals, activating caspases and other cell death execution events by releasing apoptogenic proteins from the intermembrane space. The BCL-2 family of proteins localize (or can be targeted) to mitochondria and regulate the permeability of the mitochondrial outer membrane to these apoptotic factors. Recent evidence suggests that multiple mechanisms may regulate the release of mitochondrial factors, some of which depend on the action of caspases. PMID- 15530778 TI - Death by design: apoptosis, necrosis and autophagy. AB - Apoptosis is the principal mechanism by which cells are physiologically eliminated in metazoan organisms. During apoptotic death, cells are neatly carved up by caspases and packaged into apoptotic bodies as a mechanism to avoid immune activation. Recently, necrosis, once thought of as simply a passive, unorganized way to die, has emerged as an alternate form of programmed cell death whose activation might have important biological consequences, including the induction of an inflammatory response. Autophagy has also been suggested as a possible mechanism for non-apoptotic death despite evidence from many species that autophagy represents a survival strategy in times of stress. Recent advances have helped to define the function of and mechanism for programmed necrosis and the role of autophagy in cell survival and suicide. PMID- 15530779 TI - Targeting cell cycle and apoptosis for the treatment of human malignancies. AB - Oncogenic transformation leads to cell cycle aberration and apoptosis dysregulation. Targeting cell cycle and apoptosis pathways has emerged as an attractive approach for the treatment of cancer. The activity of cdks can be modulated by targeting these kinases with small molecules that bind to the ATP binding pocket of cdks, or by altering the composition of the cdk/endogenous cdk inhibitor complexes by different mechanisms. Apoptosis can be modulated by targeting pro-apoptotic or pro-survival pathways. Several proteins relevant to oncogenic and proliferative processes, such as p53, bcl-2, AKT, ras and epidermal growth factor receptor, are also important in blocking apoptosis. Several small molecules that modulate cell cycle control and apoptosis have been approved recently and many will be approved in the near future. Several challenges remain, including finding ways of targeting these agents specifically to tumors (sparing normal cells), and the development of rationales for combining these new agents with standard therapies and for prioritizing the development of an overwhelming number of novel small molecules targeting cell cycle and apoptosis. Novel technologies such as genomics and proteomics will be instrumental in designing combinatorial regimens tailored to patients on the basis of the genetic makeup of tumors. Irrespective of all shortcomings, the future of modulation of apoptosis and cell cycle machinery for oncology therapy is quite exciting. PMID- 15530777 TI - Misfolded proteins, endoplasmic reticulum stress and neurodegeneration. AB - The accumulation of misfolded proteins (e.g. mutant or damaged proteins) triggers cellular stress responses that protect cells against the toxic buildup of such proteins. However, prolonged stress due to the buildup of these toxic proteins induces specific death pathways. Dissecting these pathways should be valuable in understanding the pathogenesis of, and ultimately in designing therapy for, neurodegenerative diseases that feature misfolded proteins. PMID- 15530781 TI - Wnt signaling and the regulation of stem cell function. AB - Canonical Wnt signaling plays a crucial role in controlling cell expansion in many types of stem cells. Recent studies, however, demonstrated that Wnt is not only a general stem cell growth factor but can also influence cell lineage decisions in certain stem cell types by promoting specific fates at the expense of others. Thus, Wnt signaling elicits multiple functions in stem cells. Wnt activity appears to depend on cell-intrinsic properties that might change with time during development, thereby altering the cellular response to Wnt. Moreover, the spatial context of a stem cell also determines how the cell interprets Wnt signal activity, in that synergistic or antagonistic signaling pathways can modulate Wnt signaling. How a stem cell integrates Wnt and other signals and how such signaling networks regulate stem cell function on the molecular level remains to be elucidated. PMID- 15530782 TI - In vitro gametogenesis from embryonic stem cells. AB - Many insights into mammalian germ cell development have been gained through genetic engineering and in vivo studies, but the lack of an in vitro system for deriving germ cells has hindered potential advances in germ cell biology. Recent studies have demonstrated embryonic stem cell differentiation into germ cells and more mature gametes, although significant unanswered questions remain about the functionality of these cells. The derivation of germ cells from embryonic stem cells in vitro provides an invaluable assay both for the genetic dissection of germ cell development and for epigenetic reprogramming, and may one day facilitate nuclear transfer technology and infertility treatments. PMID- 15530783 TI - The stem cell niche: theme and variations. AB - Stem cells in animal tissues are often located and controlled by special tissue microenvironments known as niches. Studies of stem cell niches in model systems such as Drosophila have revealed adhesive interactions, cell cycle modifications and intercellular signals that operate to control stem cell behavior. Candidate niches and regulatory molecules have also been identified in many mammalian tissues, including bone marrow, skin, gut and brain. While niches are an ancient evolutionary device with conserved features across diverse organisms, we suggest that certain niches display important differences in their organization and function. PMID- 15530784 TI - Diverse mechanisms regulate stem cell self-renewal. AB - To what extent are the pathways that regulate self-renewal conserved between stem cells at different stages of development and in different tissues? Some pathways play a strikingly conserved role in regulating the self-renewal of diverse stem cells, whereas other pathways are specific to stem cells in certain tissues or at certain stages of development. Recent studies have highlighted differences between the self-renewal of embryonic, fetal and adult stem cells. By understanding these similarities and differences we may come to a molecular understanding of how stem cells replicate themselves and why aspects of this process differ between stem cells. PMID- 15530785 TI - Cancer stem cell biology: from leukemia to solid tumors. AB - The biology of stem cells and their intrinsic properties are now recognized as integral to tumor pathogenesis in several types of cancer. This observation has broad ramifications in the cancer research field and is likely to impact our understanding of the basic mechanisms of tumor formation and the strategies we use to treat cancers. A role for stem cells has been demonstrated for cancers of the hematopoietic system, breast and brain. Going forward it is likely that stem cells will also be implicated in other malignancies. Hence, a detailed understanding of stem cells and how they mediate tumor pathogenesis will be critical in developing more effective cancer therapies. PMID- 15530786 TI - The clinical potential of stem cells. AB - Stem cells are defined by their capacity for self-renewal and multilineage differentiation, making them uniquely situated to treat a broad spectrum of human diseases. For example, because hematopoietic stem cells can reconstitute the entire blood system, bone marrow transplantation has long been used in the clinic to treat various diseases. Similarly, the transplantation of other tissue specific stem cells, such as stem cells isolated from epithelial and neural tissues, can treat mouse disease models and human patients in which epithelial and neural cells are damaged. An alternative to tissue-specific stem cell therapy takes advantage of embryonic stem cells, which are capable of differentiating into any tissue type. Furthermore, nuclear transfer, the transfer of a post mitotic somatic cell nucleus into an enucleated oocyte, creates a limitless source of autologous cells that, when combined with gene therapy, can serve as a powerful therapeutic tool. PMID- 15530787 TI - Editorial. PMID- 15530789 TI - Imaging immobilised ssDNA and detecting DNA hybridisation by means of the repelling mode of scanning electrochemical microscopy (SECM). AB - The supposed repelling mode of scanning electrochemical microscopy (SECM) allows truly label-free electrochemical recognition of the presence and hybridisation of nucleic acids that are immobilised on conducting DNA chips. Basically, the SECM based detection of single- and double-stranded DNA profits from the electrostatic repulsion between deprotonated phosphate groups at the backbone of the oligonucleotides and a free-diffusing negatively charged redox mediator (e.g. [Fe(CN)(6)](3-/4-)). In electrolytes of proper pH and ionic strength, this coulomb interaction is heavily influencing the diffusion properties of the mediator in the vicinity of the surface-anchored DNA strands. This charge interaction modulates the diffusional mass transport for the charged redox species in the DNA modified regions, and thus locally decreases the positive feedback currents measured with a SECM tip placed within the electrochemical nearfield of the chip surface. This approach was used to study arrays of synthetic 20-base oligonucleotide probes that were immobilised on monolayer modified gold surfaces. Evidence is provided that the density of probes, the ionic strength of solution and the tip-to-sample distance have a strong impact on the capability of the repelling mode of SECM to visualise probe spots and hybridisation while the concentration of the chosen mediator did not significantly affect detection. PMID- 15530788 TI - Examination of surface-bound Ku-DNA complexes in an aqueous environment using MAC mode atomic force microscopy. AB - In the development of biosensors, it is essential to understand how the signal transducing element may perturb surface-bound proteins and nucleic acids. The tip of the atomic force microscope is such an element in atomic force microscopy. In this paper, we describe the influence of tip-sample interactions on the measured height of the DNA repair protein, Ku, that has been adsorbed onto a mica surface which was submerged in aqueous solution. We find that the measured height of the Ku molecule depends critically on whether or not it is associated with DNA. Additionally, we observed that the conditions (time and concentration) under which Ku is incubated with DNA, affect the appearance (number and type) of the DNA-Ku complexes observed. PMID- 15530790 TI - AFM and electroanalytical studies of synthetic oligonucleotide hybridization. AB - The first and most important step in the development and manufacture of a sensitive DNA-biosensor for hybridization detection is the immobilization procedure of the nucleic acid probe on the transducer surface, maintaining its mobility and conformational flexibility. MAC Mode AFM images were used to demonstrate that oligonucleotide (ODN) molecules adsorb spontaneously at the electrode surface. After adsorption, the ODN layers were formed by molecules with restricted mobility, as well as by superposed molecules, which can lead to reduced hybridization efficiency. The images also showed the existence of pores in the adsorbed ODN film that revealed large parts of the electrode surface, and enabled non-specific adsorption of other ODNs on the uncovered areas. Electrostatic immobilization onto a clean glassy carbon electrode surface was followed by hybridization with complementary sequences and by control experiments with non-complementary sequences, studied using differential pulse voltammetry. The data obtained showed that non-specific adsorption strongly influenced the results, which depended on the sequence of the ODNs. In order to reduce the contribution of non-specific adsorbed ODNs during hybridization experiments, the carbon electrode surface was modified. After modification, the AFM images showed an electrode completely covered by the ODN probe film, which prevented the undesirable binding of target ODN molecules to the electrode surface. The changes of interfacial capacitance that took place after hybridization or control experiments showed the formation of a mixed multilayer that strongly depended on the local environment of the immobilized ODN. PMID- 15530791 TI - The application of ultrasound as a rapid method to provide DNA fragments suitable for detection by DNA biosensors. AB - Contamination of food and water supplies by microorganisms such as Escherichia coli, the need for point-of-care bedside analysis of biological samples, and concerns about terrorist attacks using biological organisms, have made the development of fast, reliable, and sensitive analytical methodologies for use in monitoring of pathogens very important. With a variety of biosensors being developed for extremely sensitive and rapid nucleic acid diagnostics, it has become even more important to shift focus towards creation of methods to decrease the amount of time and effort necessary for sample preparation. The application of ultrasound has the potential to create DNA fragments from genomic material with lengths that are suitable for determination using biosensors and microarrays. For example, application of 85 W power at a frequency of 20 kHz can produce a preponderance of fragments of 100-400 base pairs (bp) within several seconds, and sample processing can lead to over 75% conversion from genomic material to fragments in times of 20-30 s. A proportion of these fragments are in a single-stranded state and are suitable for hydridization with immobilized single-stranded DNA probe oligonucleotides using a fiber optic biosensor. Control of factors such as salt concentration, exposure time, ultrasound power, and the initial temperature of the solution, can affect the length and form (single- or double-stranded) of DNA fragments that are generated by ultrasound, and average fragment length can be adjusted by selection of these operating parameters. PMID- 15530792 TI - Cohort analysis of a single nucleotide polymorphism on DNA chips. AB - A method has been developed to determine SNPs on DNA chips by applying a flow through bioscanner. As a practical application we demonstrated the fast and simple SNP analysis of 24 genotypes in an array of 96 spots with a single hybridisation and dissociation experiment. The main advantage of this methodical concept is the parallel and fast analysis without any need of enzymatic digestion. Additionally, the DNA chip format used is appropriate for parallel analysis up to 400 spots. The polymorphism in the gene of the human phenol sulfotransferase SULT1A1 was studied as a model SNP. Biotinylated PCR products containing the SNP (The SNP summary web site: ) (mutant) and those containing no mutation (wild-type) were brought onto the chips coated with NeutrAvidin using non-contact spotting. This was followed by an analysis which was carried out in a flow-through biochip scanner while constantly rinsing with buffer. After removing the non-biotinylated strand a fluorescent probe was hybridised, which is complementary to the wild-type sequence. If this probe binds to a mutant sequence, then one single base is not fully matching. Thereby, the mismatched hybrid (mutant) is less stable than the full-matched hybrid (wild-type). The final step after hybridisation on the chip involves rinsing with a buffer to start dissociation of the fluorescent probe from the immobilised DNA strand. The online measurement of the fluorescence intensity by the biochip scanner provides the possibility to follow the kinetics of the hybridisation and dissociation processes. According to the different stability of the full-match and the mismatch, either visual discrimination or kinetic analysis is possible to distinguish SNP-containing sequence from the wild-type sequence. PMID- 15530793 TI - Immobilisation of DNA probes for the development of SPR-based sensing. AB - An immobilisation procedure based on the direct coupling of thiol-derivatised oligonucleotide probes to bare gold sensor surfaces has been used for DNA sensing applications. The instrumentation used relies on surface plasmon resonance (SPR) transduction; in particular the commercially available instruments BIACORE X and SPREETA, have been employed in this study. The performances of the SPR-based DNA sensors resulting from direct coupling of thiol-derivatised DNA probes onto gold chips, have been studied in terms of the main analytical parameters, i.e. selectivity, sensitivity, reproducibility, analysis time, etc. A comparison between the thiol-derivatised immobilisation approach and a reference immobilisation method, based on the coupling of biotinylated oligonucleotide probes onto a streptavidin coated dextran sensor surface, using synthetic complementary oligonucleotides has been discussed. Finally, a denaturation method to obtain ssDNA ready for hybridisation analysis has been applied to polymerase chain reaction (PCR) amplified samples, for the detection of genetically modified organisms (GMOs). PMID- 15530794 TI - Electrochemical sensing of the behaviour of oligonucleotide lipoplexes at charged interfaces. AB - Complexes between short oligodeoxynucleotides (ODN) with a variable dG(x)dC(y) base composition and liposomes composed of the cationic lipid DOTAP (ODN lipoplexes) were studied by differential pulse voltammetry at a glassy carbon electrode. Since lipoplexes are spontaneously formed by electrostatic interactions, the objective of the voltammetric study was to investigate their behaviour at the electrode surface/solution interface. It was verified that the peak current in the voltammograms for ODN lipoplexes was due to guanosine oxidation and that it was influenced both by the applied adsorption potential and the lipoplex (+/-) charge ratio used. It was found that for low ODN lipoplexes (+/-) charge ratios the peak current obtained was enhanced when compared to that registered with free ODN for the same concentration. This allowed a higher sensitivity in the determination of ODN by differential pulse voltammetry and a limit of detection of 5.5 ng/mL was achieved. A model that explains the organisation of ODN lipoplexes at the electrode surface/solution interface is proposed. The electrochemical results presented account for a better physicochemical characterisation of lipoplexes at charged interfaces, which can be important for the understanding and development of gene therapy vectors based on ODN lipoplexes. PMID- 15530795 TI - Multiply osmium-labeled reporter probes for electrochemical DNA hybridization assays: detection of trinucleotide repeats. AB - In electrochemical DNA hybridization assays target or probe DNAs end-labeled with electroactive compounds have been frequently used. We show that multiple osmium labels yielding faradaic (at carbon or mercury electrodes) and catalytic signals (at mercury electrodes) can be easily covalently bound to DNA molecules. We use (GAA)(7) (T)(n) oligodeoxynucleotides (ODNs) with n ranging between 5 and 50. (T)(n) tails are selectively modified with osmium tetroxide,2,2'-bipyridine leaving the (GAA)(7) repeat intact for the DNA hybridization. These ODNs are applied as reporter probes (RP's) in DNA hybridization double-surface (DS) assay using magnetic beads for the DNA hybridization and pyrolytic graphite (PGE) or hanging mercury drop (HMDE) electrodes for the electrochemical detection. We show that in difference to the usual single-surface methods (where the RP has to be bound to target DNA near to the surface to communicate with the electrode) in the DS assay the RP can be bound to DNA regardless of its position and can used for the determination of the length of DNA repetitive sequences. Several fmols or about a hundred of amol of a RP with osmium-labeled (T)(50) tail can be detected at PGE and HMDE, respectively, at 1-2 min accumulation time. PMID- 15530796 TI - Carbon-nanotube-modified electrodes for amplified enzyme-based electrical detection of DNA hybridization. AB - Carbon-nanotube-modified glassy carbon (CNT/GC) transducers have been developed for enhancing the sensitivity and stability of enzyme-based electrochemical bioassays of DNA hybridization. The amplified signal reflects the interfacial accumulation of phenolic products of the alkaline-phosphatase tracer onto the CNT layer. In particular, chronopotentiometric measurements (following short accumulation periods) offer a substantial enhancement of the response of enzymatically liberated alpha-naphthol. The CNT modifier also leads to a dramatic improvement in the stability of the amperometric response of alpha-naphthol. These advantages of CNT/GC electrodes are illustrated from comparison to unmodified glassy carbon electrodes. Factors influencing the adsorptive accumulation of alpha-naphthol, and the overall performance of the new DNA assay, are assessed and optimized. The attractive performance characteristics of the new multi-amplification electrochemical detection of DNA hybridization are reported in connection to the detection of nucleic acid sequences related to the breast cancer BRCA1 gene. PMID- 15530797 TI - A strand exchange FRET assay for DNA. AB - A new displacement hybridisation method is reported using a single strand DNA probe, labelled with an acceptor fluorophore (oregon green 488). Detection of double stranded sample target is shown, with discrimination between the probe, duplexed during the assay, and free single stranded probe DNA achieved through the FRET from a donor grove fluorophore (Hoechst 33258). A model for the kinetics of the displacement assay is presented and the course of the assay predicted according to probe/target ratios and sequence. The modelled predictions are consistent with the experimental data showing single base pair mismatch discrimination. The pattern of response according to the mismatch/perfect complement ratio in a mixed sample is also considered with an allele discrimination ratio lying between the homozygous gene and total mismatch case, according to ratio. The assay is shown to be tolerant of different probe concentrations and ratios and through the dual wavelength recorded signals from donor and FRET acceptor, internal baseline correction is achieved with excellent noise reduction through ratiometric measurement. PMID- 15530798 TI - Amplified detection of telomerase activity using electrochemical and quartz crystal microbalance measurements. AB - Telomerase is considered as an important biomarker for cancer cells. Two different methods for the amplified electrochemical and microgravimetric quartz crystal-microbalance detection of telomerase activity originating from HeLa cancer cells are described. One method involves the telomerization of a primer (1) linked to the electrode, in the presence of telomerase from HeLa cell extract and dNTP, followed by the hybridization of a biotin-labeled nucleic acid (2) that is complementary to the telomere repeat units. The subsequent binding of an avidin-alkaline phosphatase conjugate (3) that catalyzes the oxidative hydrolysis of 5-bromo-4-chloro-3-indolyl phosphate (4) results in the precipitation of the insoluble product (5) on the electrode. The second method involves the telomerization of the primer (1) associated with the electrode, in the presence of the telomerase-containing HeLa cell extract and the dNTP nucleotide mixture that includes biotin-labeled dUTP. The telomerization leads to the labeling of the telomeres with biotin labels. The association of the avidin-alkaline phosphatase conjugate (3) to the biotin labels results in the biocatalyzed transformation of (4) to (5) and the formation of a precipitate on the electrode or the Au-quartz crystal. As numerous precipitate molecules are formed as a result of the formation of a single telomere, the methods represent routes for the amplified detection of telomerase activity. The formation of the precipitate on the respective transducers is probed by following the changes in the electrode resistance using chronopotentiometry, or by following the frequency changes of the piezoelectric quartz crystals. The amount of precipitate generated on the electrodes is controlled by the concentration of the HeLa cancer cells. The methods enable the detection of telomerase activity that is extracted from 1000 HeLa cancer cells. PMID- 15530799 TI - Sensors for toxicity of chemicals and oxidative stress based on electrochemical catalytic DNA oxidation. AB - Films of DNA, enzymes, polyions, and catalytic redox polyions of nanometer thickness on electrodes can provide active elements for sensors for screening the toxicity of chemicals and their metabolites, and for oxidative stress. The unifying feature of this approach involves layer-by-layer electrostatic assembly of films designed to detect DNA damage. Films containing DNA and enzymes enable detection of structural damage to DNA as a basis for toxicity screening. These films bioactivate chemicals to their metabolites, which can then react with DNA, mimicking toxicity pathways in the human liver. Metallopolyions that catalyze DNA oxidation can be incorporated into DNA/enzyme films leading to "reagentless" sensors. These sensors are suitable for detecting relative DNA damage rates in <5 min of the enzyme reactions. Films of the osmium polymer [Os(bpy)(2)(PVP)(10)Cl](+) [poly(vinylpyridine), PVP] can be used to monitor DNA oxidation selectively. Such films may be applicable to determination of oxidized DNA as a clinical biomarker for oxidative stress. Inclusion of the analogous ruthenium metallopolymer in the sensor provides a monitor for oxidation of other nucleobases. PMID- 15530800 TI - Depressing? Heck no, uplifting! PMID- 15530801 TI - The effect of varying user-selected input parameters on quantitative values in CT perfusion maps. AB - RATIONALE AND OBJECTIVES: Deconvolution-based software can be used to calculate quantitative maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) from first-pass computed tomography perfusion (CTP) datasets. The application of this software requires the user to select multiple input variables. The purpose of this study was to investigate the degree to which both major and minor variations of these user-defined inputs would affect the final quantitative values of CBF, CBV, and MTT. MATERIALS AND METHODS: A neuroradiologist constructed CBF, CBV, and MTT maps using standard methodology with commercially available software (GE Functool Version 1.9s) from CTP datasets of three acute stroke patients. Each map was reconstructed multiple times by systematically and independently varying the following parameters: postenhancement and preenhancement cutoff values, arterial and venous region-of interest (ROI) placement, and arterial and venous ROI size. The resulting quantitative CTP values were compared using identical ROIs placed at the infarct core. RESULTS: Major variations of either arterial ROI placement or arterial and venous ROI size had no significant effect on the mean CBF, CBV, and MTT values at the infarct core (p > .05). Even minor variations, however, in the choice of venous ROI placement or in pre- and postenhancement cutoff values significantly altered the quantitative values for each of the CTP maps, by as much as threefold. CONCLUSION: Even minor variations of user-defined inputs can significantly influence the quantitative, deconvolution-based CTP map values of acute stroke patients. Although quantitation was robust to the choice of arterial ROI placement and arterial or venous ROI size, it was strongly dependent on the choice of venous ROI location and pre- and postenhancement cut-off values. Awareness of these results by clinicians may be important in the creation of quantitatively accurate CTP maps. PMID- 15530803 TI - Post-reconstruction filtering of positron emission tomography whole-body emission images and attenuation maps using nonlinear diffusion filtering. AB - RATIONALE AND OBJECTIVES: Positron emission tomography has been playing an important role as a quantitative molecular imaging modality to measure and image biochemical processes in vivo. The quality of positron emission tomography images may impact the accuracy of the quantitative or semiquantitative information extracted from them. MATERIALS AND METHODS: In this study, the anisotropic diffusion filtering technique was used to de-noise emission images and attenuation maps of a whole body for improving the quantitative accuracy of emission images. The efficacy of the filtering technique was shown on a hot lesion phantom as well as on whole-body emission images and attenuation maps. RESULTS: The filtering technique showed a superb performance on images reconstructed using the iterative and the filtered back-projection reconstruction techniques. CONCLUSION: Filtering may allow the reconstruction of images with optimal parameters with respect to lesion contrast and image resolution by removing the consequential noise. An improvement of as much as six-fold may be attained. It may also improve the accuracy of the quantitative information, such as standard uptake value extracted from emission images. PMID- 15530802 TI - Automated computerized scheme for detection of unruptured intracranial aneurysms in three-dimensional magnetic resonance angiography. AB - RATIONALE AND OBJECTIVES: A computerized scheme for automated detection of unruptured intracranial aneurysms in magnetic resonance angiography was developed based on the use of a three-dimensional selective enhancement filter for dots (aneurysms). MATERIALS AND METHODS: Twenty-nine cases with 36 unruptured aneurysms (diameter, 3 to 26 mm; mean, 6.6 mm) and 31 non-aneurysm cases were used in this study. The isotropic 3-dimensional magnetic resonance angiography images with 400 x 400 x 128 voxels (voxel size, 0.5 mm) were processed by use of the selective enhancement filter. The initial candidates were identified by use of a multiple gray-level thresholding technique on the dot-enhanced images and a region-growing technique with monitoring some image features. All candidates were classified into four types of candidates according to the size and local structures based on the effective diameter and skeleton image of each candidate (ie, large candidates and three types of small candidates including short-branch type, single-vessel type, and bifurcation type). In each group, a number of false positives were removed by use of different rules on localized image features related to gray levels and morphology. Linear discriminant analysis was used for further removal of false-positives. RESULTS: With this computer-aided diagnostic scheme, all of 36 aneurysms were correctly detected with 2.4 false-positives per patient based on a leave-one-out-by-patient test method. CONCLUSION: This computer-aided diagnostic system would be useful in assisting radiologists for the detection of intracranial aneurysms in magnetic resonance angiography. PMID- 15530804 TI - Combined use of MRI and PET to monitor response and assess residual disease for locally advanced breast cancer treated with neoadjuvant chemotherapy. AB - RATIONALE AND OBJECTIVES: The purpose of the study was to evaluate the hypothesis that magnetic resonance imaging (MRI) and positron emission tomography (PET) are complementary and valuable in monitoring response and assessing residual disease of locally advanced breast cancer (LABC) treated with neoadjuvant chemotherapy. We sought to determine if the combination of the two modalities was more accurate than either alone and could provide better guidance in patient management. MATERIALS AND METHODS: Sixteen lesions in 15 women with LABC were evaluated with MRI, PET, and clinical breast examination (CBE) before and after neoadjuvant chemotherapy. The pre- and posttherapy maximal tumor sizes on MRI and CBE and standard uptake values (SUVs) on PET served as the measurements for clinical response classification and residual disease assessment. Pathologic assessment provided the reference for macroscopic and microscopic pathologic tumor response and residual disease. RESULTS: PET correctly predicted lack of pathologic response in five of six cases (83%); CBE predicted correctly in one of six (17%) cases, and MRI predicted correctly in zero of six cases. When PET predicted response, MRI defined the extent of macroscopic pathologic residual disease accurately in 9 of 10 cases (90%). When posttherapy MRI showed complete response (CR) in eight cases, macroscopic pathologic complete response (mCR) was observed in all eight cases (100%). CONCLUSION: Our study suggests that combined use of MRI and PET is complementary and offers advantages over CBE. PET was more accurate in predicting pathologic nonresponse. Complete response by MRI correlated well with macroscopic pathologic complete response. PMID- 15530805 TI - Segmentation of tumors in magnetic resonance brain images using an interactive multiscale watershed algorithm. AB - RATIONALE AND OBJECTIVE: This article presents the evaluation of an interactive multiscale watershed segmentation algorithm for segmenting tumors in magnetic resonance brain images of patients scheduled for neuronavigational procedures. MATERIALS AND METHODS: The watershed method is compared with manual delineation with respect to accuracy, repeatability, and efficiency. RESULTS: In the 20 patients included in this study, the measured volume of the tumors ranged from 2.7 to 81.9 cm(3). A comparison of the tumor volumes measured with watershed segmentation to the volumes measured with manual delineation shows that the two methods are interchangeable according to the Bland and Altman criterion, and thus equally accurate. The repeatability of the watershed method and the manual method are compared by looking at the similarity of the segmented volumes. The similarity for intraobserver and interobserver variability for watershed segmentation is 96.4% and 95.3%, respectively, compared with 93.5% and 90.0% for manual outlining, from which it may be concluded that the watershed method is more repeatable. Moreover, the watershed algorithm is on average three times faster than manual outlining. CONCLUSION: The watershed method has an accuracy comparable to that of manual delineation and outperforms manual outlining on the criteria of repeatability and efficiency. PMID- 15530806 TI - Computer-aided detection for screening mammography. AB - Mammographic film reading is a highly demanding task, particularly in screening programs where the reader must perform a detailed visual search of a large number of images for signs of abnormality that are often subtle or small, and which occur very infrequently. False-negative cases, in which signs of abnormality are missed by a film reader, are known to occur. Computer-aided detection (CAD) systems, which automatically detect potential abnormalities and indicate their locations to the reader, have the capacity to reduce the frequency of such errors by ensuring that all suspicious regions of the images are thoroughly searched and by increasing the weighting attached to subtle signs that may otherwise have been dismissed. CAD systems depend on suites of detection algorithms, but each algorithm has a different sensitivity and specificity and the effect of prompting errors on human performance with CAD is complex. This article is a brief review of CAD for screening mammography; it highlights both the strengths and the weaknesses of the approach, and describes some of the methodologies used to evaluate CAD in a clinical setting. PMID- 15530807 TI - New opportunities for the use of digital video in on-line radiologic curricula. AB - RATIONALE AND OBJECTIVES: To explore techniques to conveniently and self sufficiently create high-quality, web-ready instructional digital video snippets suitable for routine use in on-line radiologic curricula. MATERIALS: A commercially available digital camera with an 8-megapixel image receptor and the capability to record web-ready digital video and audio at a resolution of up to 640 x 480 pixels and a frame rate of up to 30 per second was used to obtain video snippets intended for inclusion in on-line curricula. Hand-held and tripod techniques were compared. Evaluation focused on the types of snippets deemed most likely to be used within on-line educational content, ranging in length from 10 seconds to 2 minutes. Additionally, basic postprocessing functions to experiment with combinations of video-file size, format, transmission efficiency, and image quality were used. RESULTS: The overall video quality was considered by participating radiologists to be good to excellent for its intended purposes. For most situations tested, a matrix size of 320 x 240 pixels provided a good balance of visual quality versus file size and transmission overhead. The 640 x 480 format was occasionally optimal, but was usually larger than necessary and resulted in substantially larger files, especially at a rate of 30 frames per second. A rate of 15 frames per second was considered adequate for most situations, regardless of matrix size, although it did add some barely discernible choppiness to the display. For all but very short snippets, a tripod or some other form of stabilization was necessary to eliminate distracting "camera shake" associated with free-hand acquisition. With a tripod, appropriate ambient lighting, and a favorable acoustical environment, videos of excellent quality when using the highest settings available on the camera (640 x 480, 30 frames/sec) could be self-sufficiently produced. However, such high-quality video with its associated large file size was rarely the optimal fit for the needs of this curricula. CONCLUSION: These results indicate that newer-generation digital cameras are useful for quickly and inexpensively producing high-quality, web ready digital video suitable for use in on-line education. PMID- 15530808 TI - Comprehensive innovative solution for resident education using the Intranet Journal of Chest Radiology. AB - RATIONALE AND OBJECTIVES: Because of the rapid increase in clinical workload in academic radiology departments, time for teaching rotating residents is getting more and more limited. As a solution to this problem, we introduced the Intranet Journal of Chest Radiology as a comprehensive innovative tool for assisting resident education. MATERIALS AND METHODS: The Intranet Journal of Chest Radiology is constructed using Microsoft FrontPage version 2002 (Microsoft Corp, Redmond, WA) and is hosted in our departmental web server (Beth Israel Deaconess Medical Center, Boston, MA). The home page of the intranet journal provides access to the main features, "Cases of the Month," "Teaching File," "Selected Articles for Residents," "Lecture Series," and "Current Publications." These features provide quick access to the selected radiology articles, the interesting chest cases, and the lecture series and current publication from the chest section. RESULTS: Our intranet journal has been well utilized for 6 months after its introduction. It enhances residents' interest and motivation to work on case collections, to search and read articles, and to generate interest in research. Frequent updating is necessary for the journal to be kept current, relevant, and well-utilized. CONCLUSION: The intranet journal serves as a comprehensive innovative solution for resident education, providing basic educational resources and opportunities of interactive participation by residents. PMID- 15530809 TI - Use of personal digital assistants in diagnostic radiology resident education. AB - RATIONALE AND OBJECTIVES: Personal digital assistants (PDAs) are gaining widespread use in the medical community. We introduced a PDA-based mobile system that provides departmental and educational information with a seamless connection to the intranet. The objective of this study is to determine the impact a PDA has on educational resources (learning or data reference) brought to work and used at home by a radiology resident based on user surveys. MATERIALS AND METHODS: Survey was performed on 32 radiology residents in our department before and 6 months after the release of the PDA-based system. We assessed the changes in (1) sources of learning at home and at work, and in (2) data reference. The second survey also evaluated the usefulness of each component of the system. RESULTS: After the release of the PDA-based mobile system, the use of "digital books and references" as data references and educational resources that were brought to work every day significantly increased (P = .016, P < .0001, respectively). "Traditional books and references" remained the "most useful source in learning radiology"; however, "digital books and references" increased as the residents' first choice from 0% to 16% within 6 months of introducing the package (P = .125). CONCLUSION: The introduction of a PDA-based system consisting of educational and departmental information had a statistically significant impact in increasing the use of digitized information in radiology resident education. PMID- 15530810 TI - A primer on molecular biology for imagers: VIII. Equipment for imaging molecular processes. PMID- 15530811 TI - A small animal model of regional alveolar ventilation using HP 3He MRI1. AB - RATIONALE AND OBJECTIVES: The aim of this study was to establish a standardized procedure for the measurement of regional fractional ventilation in a healthy rat model as a baseline for further studies of pulmonary disorder models. MATERIALS AND METHODS: The lungs of five healthy male Sprague-Dawley rats were imaged using hyperpolarized helium-3 magnetic resonance imaging. From these images, regional fractional ventilation was calculated and maps generated detailing the distribution of fractional ventilation in the lung. The 1.56 mm x 1.56 mm x 4 mm regions of interest were assigned on 5 cm x 5 cm field of view lung maps. Histograms were also generated showing the frequency distribution of fractional ventilation values. To compare fractional ventilation values between animals, the ventilation procedure was standardized to results from individual pulmonary function tests. Each animal's spontaneous tidal volume, respiratory rate, and inspiration percentage (percent of total respiratory cycle in inspiration) were used in their mechanical ventilation settings. RESULTS: Results were similar among all five healthy rats based on examination of ventilation distribution maps and frequency distribution histograms. Mean (0.13) and standard deviation (0.07) were calculated for fractional ventilation in each animal. However, these values were determined to be influenced by slice selection, and therefore the maps and histograms were favored in analysis of results. CONCLUSION: This study shows consistent results in healthy rat lungs and will serve as a baseline study for future measurements in emphysematous rat lungs. PMID- 15530812 TI - MR appearances of radiofrequency thermal ablation region: histopathologic correlation with dog liver models and an autopsy case. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate the utility of magnetic resonance (MR) imaging for indicating the extent of true tissue necrosis of the liver after radiofrequency (RF) ablation in comparison with histopathologic findings in dog models and an autopsy case. MATERIALS AND METHODS: RF ablation of the liver parenchyma was performed on three dogs under general anesthesia. MR appearances of the RF-ablated regions on T1-weighted fast low angle shot (FLASH; repetition time/echo time [TR/TE]/flip angle: 120/3.8/70),T2-weighted turbo spin echo (3000/80/echo train = 25) and contrast enhanced T1-weighted images were compared with histopathologic findings. An autopsy case with hepatocellular carcinoma was also enrolled in this study. RESULTS: All ablated regions showed three zones on T1-weighted FLASH images: a central zone with low intensity, a broad hyperintense middle zone, and a surrounding hypointense band. The central and middle zones corresponded to the degrees of coagulation necrosis observed during histopathologic examination, whereas no viable cells were seen in these zones during the microscopic examination using nicotinamide adenine dinucleotide diaphorase stain. The surrounding hypointense band corresponded to sinusoidal congestion in the acute phase and to fibrotic change in the subacute phase. CONCLUSION: MR imaging using the FLASH sequence can accurately determine the extent of the necrotic area after RF ablation. PMID- 15530813 TI - Chasing contrast molecules: a 45-year quixotic quest. PMID- 15530814 TI - Reporting instruction for radiology residents. PMID- 15530817 TI - Hemoglobin-binding protein HgbA in the outer membrane of Actinobacillus pleuropneumoniae: homology modelling reveals regions of potential interactions with hemoglobin and heme. AB - Analyses of the primary sequence of hemoglobin-binding protein HgbA from Actinobacillus pleuropneumoniae by comparative modelling and by a Hidden Markov Model identified its topological similarities to bacterial outer membrane receptors BtuB, FepA, FhuA, and FecA of Escherichia coli. The HgbA model has a globular N-terminal cork domain contained within a 22-stranded beta barrel domain, its folds being similar to the structures of outer membrane receptors that have been solved by X-ray crystallography. The barrel domain of the HgbA model superimposes onto the barrel domains of the four outer membrane receptors with rmsd values less than 1.0 A. This feature is consistent with a phylogenetic tree which indicated clustering of polypeptide sequences for three barrel domains. Furthermore, the HgbA model shares the highest structural similarity to BtuB, with the modelled HgbA barrel having approximately the same elliptical cross-section and height as that of BtuB. Extracellular loop regions of HgbA are predicted to be more extended than those of the E. coli outer membrane receptors, potentially facilitating a protein-protein interface with hemoglobin. Fold recognition modelling of the HgbA loop regions showed that 10 out of 11 predicted loops are highly homologous to known structures of protein loops that contribute to heme/iron or protein-protein interactions. Strikingly, HgbA loop 2 has structural homology to a loop in bovine endothelial nitric acid oxidase that is proximal to a heme-binding site; and HgbA loop 7 contains a histidine residue conserved in a motif that is involved in heme/hemoglobin interactions. These findings implicate HgbA loops 2 and 7 in recognition and binding of hemoglobin or the heme ligand. PMID- 15530818 TI - Structure and dynamics of a predicted ferredoxin-like selenoprotein in Japanese encephalitis virus. AB - Homologues of the selenoprotein glutathione peroxidase (GPx) have been previously identified in poxviruses and in RNA viruses including HIV-1 and hepatitis C virus (HCV). Sequence analysis of the NS4 region of Japanese encephalitis virus (JEV) suggests it may encode a structurally related but functionally distinct selenoprotein gene, more closely related to the iron-binding protein ferredoxin than to GPx, with three highly conserved UGA codons that align with essential Cys residues of ferredoxin. Comparison of the probe JEV sequence to an aligned family of ferredoxin sequences gave an overall 30.3% identity and 45.8% similarity, and was statistically significant at 4.9 S.D. (P < 10(-6)) above the average score computed for randomly shuffled sequences. A 3-dimensional model of the hypothetical JEV protein (JEV model) was constructed by homology modeling using SYBYL, based upon a high resolution X-ray structure of ferredoxin (PDB code: 1awd). The JEV model and the model from 1awd were subsequently subjected to molecular dynamics simulations in aqueous medium using AMBER 6. The solution structure of the JEV model indicates that it could fold into a tertiary structure globally similar to ferredoxin 1awd, with RMSD between the averaged structures of 1.8 A for the aligned regions. The modeling and MD simulations data also indicate that this structure for the JEV protein is energetically favorable, and that it could be quite stable at room temperature. This protein might play a role in JEV infection and replication via TNF and other cellular stimuli mediated via redox mechanisms. PMID- 15530819 TI - Hierarchical molecular modelling with ellipsoids. AB - Protein and DNA structures are represented at varying levels of details using ellipsoidal RGBA textured splats. The splat texture at each level is generated by rendering its children in a hierarchical model, from a distribution of viewing directions, and averaging the result. For rendering, the ellipsoids to be used are chosen adaptively, depending on the distance to the viewpoint. This technique is applied to visualize DNA coiling around nucleosomes in chromosomes. PMID- 15530820 TI - A CoMFA study of COX-2 inhibitors with receptor based alignment. AB - A diverse set of 53 cyclooxygenase-2 (COX-2) inhibitors which were aligned in two different ways were subjected to CoMFA analysis. The first method of alignment of the molecules was based on the binding information sourced from the crystallographic study, from which CoMFA Model 1 was derived. The second mode of alignment was generated by docking the inhibitors in the binding pocket using the DOCK and AFFINITY suite of programs; this gave a second model. The CoMFA Model 2 was slightly better than Model 1 in terms of the statistical parameters r(2) and q(2). The two models could predict very well the activity of a test set of diverse molecules, with a predictive r(2) of 0.593 and 0.768, respectively. Besides the QSAR results, the docking studies give a deep insight into the H bonding interactions between the inhibitors and residues in the active site of the enzyme, which can be exploited in designing better inhibitors. Useful ideas on activity improvement could be gleaned from these models. PMID- 15530821 TI - The use of a pharmacophore model for identification of novel ligands for the benzodiazepine binding site of the GABAA receptor. AB - A Catalyst pharmacophore model has been developed for the benzodiazepine site within the GABA(A) receptor complex. The model is based on a pharmacophore model originally proposed by Cook and co-workers (Drug Des. Discovery 1995, 12, 193 248) and further developed by Kahnberg et al. (J. Med. Chem. 2002, 45, 4188 4201). The Catalyst pharmacophore model has been validated by using a series of flavonoids with varying affinities for the benzodiazepine receptor and has then been used as a search query in database searching with the aim of finding novel structures which have the possibility to be modified into novel lead compounds. Five of the hits from the database searching were purchased and their affinities for the benzodiazepine site of the GABA(A) receptor were determined. Two of the compounds displayed K(i) values below 10 microM. The substance showing highest potency in-vitro displayed an affinity of 121 nM making it an interesting compound for optimization. The false positive compounds (K(i) values >10 microM affinities) have been analysed in terms of conformational energy penalties and possibilities for hydrogen bond interactions. The analysis clearly demonstrates the need for post processing of Catalyst hits. PMID- 15530822 TI - Modeling the alpha-helix to beta-hairpin transition mechanism and the formation of oligomeric aggregates of the fibrillogenic peptide Abeta(12-28): insights from all-atom molecular dynamics simulations. AB - In this paper, all-atom molecular dynamics simulations in explicit solvent are used to investigate the structural and dynamical determinants of the alpha helical to beta-hairpin conformational transition of the 12-28 fragment from the full length Abeta Alzheimer's peptide. The transition from alpha-helical to beta structure requires the peptide to populate intermediate beta-bend geometries in which several mainly hydrophobic interactions are partially formed. This is followed by the sudden collapse to ordered beta-hairpin structures and the simultaneous disruption of the hydrophobic side-chain interactions with a consequent increase in solvent exposure. The solvent exposure of hydrophobic side chains belonging to a sequence of five consecutive residues in the beta-hairpin defines a possible starting point for the onset of the aggregation mechanisms. Several different conformations of model oligomeric (dimeric and tetrameric) aggregates are then investigated. These simulations show that while hydrophobic contacts are important to bring together different monomers with a beta-hairpin like conformation, more specific interactions such as hydrogen-bonding and coulombic interactions, should be considered necessary to provide further stabilization and ordering to the nascent fibrillar aggregates. PMID- 15530823 TI - A possible model of benzimidazole binding to beta-tubulin disclosed by invoking an inter-domain movement. AB - Although it is well established that benzimidazole (BZMs) compounds exert their therapeutic effects through binding to helminth beta-tubulin and thus disrupting microtubule-based processes in the parasites, the precise location of the benzimidazole-binding site on the beta-tubulin molecule has yet to be determined. In the present study, we have used previous experimental data as cues to help identify this site. Firstly, benzimidazole resistance has been correlated with a phenylalanine-to-tyrosine substitution at position 200 of Haemonchus contortus beta-tubulin isotype-I. Secondly, site-directed mutagenesis studies, using fungi, have shown that other residues in this region of the protein can influence the interaction of benzimidazoles with beta-tubulin. However, the atomic structure of the alphabeta-tubulin dimer shows that residue 200 and the other implicated residues are buried within the protein. This poses the question: how might benzimidazoles interact with these apparently inaccessible residues? In the present study, we present a mechanism by which those residues generally believed to interact with benzimidazoles may become accessible to the drugs. Furthermore, by docking albendazole-sulphoxide into a modelled H. contortus beta-tubulin molecule we offer a structural explanation for how the mutation conferring benzimidazole resistance in nematodes may act, as well as a possible explanation for the species-specificity of benzimidazole anthelmintics. PMID- 15530825 TI - On the graphical analysis of the electronic structure of ferromagnetic clusters of medium size. AB - In a previous work, a theoretical approach, suitable to describe systems having a large number of fermions, was proposed, and results for ferromagnetic surface clusters of medium size (100-1000 atoms) were presented. The aim of the present contribution is to complete this previous work. Several significant theoretical and technical details, omitted previously, are provided here. The obtained spin orbitals are analyzed, by studying their symmetry, energy, and d contribution properties. PMID- 15530824 TI - A comparison of calculated NMR shielding probes. AB - In a strong magnetic field, covalently bonded hydrogen nuclei located over the plane of an anisotropic pi bond-containing functional group experience magnetic shielding (or deshielding) that results from the combined effect of the magnetic anisotropy of the functional group and other nearby covalent bonds plus other intramolecular shielding effects. These effects can now be calculated with reasonable accuracy using ab initio methods. We have investigated several computational probes of the magnetic shielding surface near anisotropic functional groups and compared the results to previous reports of experimental observations in example structures. GIAO-HF in Gaussian 03 was employed to calculate isotropic shielding values and to predict the net NMR shielding increment for several computational probes: methane, diatomic hydrogen, a hydrogen atom, a helium atom, or a ghost atom, each held in various positions over simple test molecules (ethene, ethyne, benzene and HCN) that contain the functional groups studied. Also, the effect of performing single point calculations versus constrained geometry-optimized calculations was examined. In addition, the effect of the angle of the orientation of the probe molecule (in the case of CH(4) and H(2)) relative to the pi bond in the test molecule was studied. Finally, the atomic charges in the molecular probes (CH(4) and H(2)) were computed to investigate the nature of the interaction of the probe with the test molecule. The optimal, most economical computational results were obtained using single point calculations of a diatomic hydrogen probe oriented perpendicular to the surface (or axis) of the test molecule. PMID- 15530826 TI - Back again to the future of Immunology Today. PMID- 15530827 TI - Looking back 25 years. PMID- 15530828 TI - Trends in Immunology--yesterday, today and tomorrow. PMID- 15530829 TI - Some early Trends in Immunology. PMID- 15530830 TI - New directions in MS therapeutics: vehicles of hope. AB - Basic immunological research has greatly expanded our understanding of the suspected immunopathology of multiple sclerosis (MS) and, more importantly, spawned a new generation of clinical trials evaluating dozens of immune-based therapies. Adhesion molecules are the furthest into development, although patient acceptance and neutralizing antibodies both support the development of small, orally available molecules. Progressive MS probably has a significant neurodegenerative component, so progress with neuroprotective strategies will require appropriate animal models, as well as more advanced clinical imaging techniques, such as brain atrophy and diffusion tensor. Dozens of therapies are in various stages of clinical development and results from these clinical studies will provide important tests of immune and inflammatory mechanisms of MS disease. PMID- 15530831 TI - 25 years since the eradication of smallpox: why poxvirus research is still relevant. AB - The World Health Organization (WHO) announced the eradication of smallpox twenty five years ago this month. This conquest of an infectious disease, which has been the bane of humankind for centuries, still stands as the WHO's greatest achievement. The anniversary of such a scientific and medical landmark provides an appropriate occasion to reflect on this feat and to assess the significance and necessity of the poxvirus research that has followed this. PMID- 15530832 TI - Evolution of innate and adaptive immunity: can we draw a line? AB - Several recent findings in the field of comparative immunology have reinforced the importance of examining the molecular and functional features of immune systems in a variety of organisms. Particularly exciting are the discoveries of a new gene rearrangement mechanism in lampreys and a somatic diversification of mollusk immune genes. These immune features being found in animals previously believed only to have innate immunity, as well as the flood of information on immune genes, molecules and mechanisms in many different creatures, have prompted us to revisit the artificial dichotomy between adaptive and innate immune systems. Although we draw no startling conclusions, we hope to encourage different thought patterns when viewing immune systems. PMID- 15530833 TI - Relationships between hematopoietic stem cells and lymphocyte progenitors. AB - Remarkable advances have been made in characterizing highly purified hematopoietic stem cells and their lymphoid progeny. The majority of the cytokines and cell interaction molecules that comprise their environment are probably known, as are most cell cycle regulatory proteins, signaling molecules and transcription factors that mediate the lineage fate decisions and control population dynamics. However, it is becoming obvious that traditional differentiation diagrams are not adequate for depicting early changes and give overly rigid impressions about how cells of the immune system arise. PMID- 15530834 TI - A central role for tissue-resident dendritic cells in innate responses. AB - In the past 20 years, dendritic cells (DCs) have been investigated mainly with regard to antigen-specific immune responses and acquired immunity. More recently, new data have indicated a novel and fundamental role for DCs in innate immunity, which is shown by the ability of DCs to induce natural killer (NK) cell activation. In mice, the molecular mechanism by which NK cells are activated by DCs has been revealed, and it consists of the induction of interleukin-2 (IL-2) expression during the early phases of an immune response. Moreover, as indicated by genome-wide transcriptional analysis, microbially activated myeloid DCs produce other chemokines and inflammatory cytokines, such as type I interferons, which in addition to IL-2 are also involved in NK cell activation. This indicates that DCs have a key role in innate immunity. The expression of an innate receptor repertoire on DCs mediates microbial recognition and uptake, and the antigen processing and specific sorting mechanisms lead to very efficient antigen presentation by DCs. Thus, we believe that DCs participate fully in innate responses, forming a bridge between innate and acquired immunity. PMID- 15530835 TI - Transfer of antigen between migrating and lymph node-resident DCs in peripheral T cell tolerance and immunity. AB - Dendritic cells (DCs) consist of a heterogeneous collection of subsets, many with unique phenotypic and functional characteristics. Although certain subsets migrate from peripheral non-lymphoid tissues, there is evidence that antigen presentation can extend to DCs that permanently reside within the lymph node. This Opinion describes this finding in the context of antigen transfer between migrating and lymphoid-resident DCs in cases of T-cell priming and tolerance induction. PMID- 15530836 TI - DCs and CD40-activated B cells: current and future avenues to cellular cancer immunotherapy. AB - Despite the still poorly understood complexity of tumor-host immune interactions, the use of cellular vaccines (particularly dendritic cells) has made it possible to reliably generate tumor antigen-specific T cells, both in animal models and in humans. These encouraging pre-clinical results have led to a translation of these immunotherapeutic strategies into clinical trials. With numerous trials still underway, their general outcome has so far been disappointing, and the discrepancy between pre-clinical data and clinical response rates is striking. Thus, either the pre-clinical models have not been representative of the human situation or the translation into human clinical trials is still sub-optimal. Here we suggest new avenues of clinical research to further improve cellular cancer immunotherapy. PMID- 15530837 TI - Historical review: Sir Gustav Nossal--immunologist and more. AB - Sir Gustav Nossal, universally known as Gus Nossal, is Australia's best known medical research scientist and advocate, who over the past five decades has influenced the course of immunology. His research interests in this field commenced during PhD studies, and his accomplishments centre around the cellular events of antibody production. His scientific achievements are matched by the energy he has devoted over his entire career to public health; science in society; political, philanthropic and community support for science; global access to existing vaccines and development of new vaccines; and the personal and scientific development of colleagues and young scientists. His contribution and philosophy of science, together with political influence, capacity to inspire and an extraordinarily engaging personality, place Gus Nossal in the top bracket of biomedical scientists of the second half of the 20th century. PMID- 15530838 TI - Different checkpoints in human NK-cell activation. AB - After the discovery, in humans and mice, of inhibitory natural killer (NK) receptors specific for MHC class I molecules, the mechanism by which NK cells kill tumor or virus-infected cells was thought to be clarified: NK cells would kill those target cells that have lost, or underexpress, MHC class I molecules. However, a more complex scenario has recently emerged. For example, certain NK cells express insufficient amounts of triggering receptors, and target cells can lack ligands for such receptors. Thus, it appears that the activation of NK cells and their potentially harmful effector functions are under the control of different checkpoints. PMID- 15530839 TI - The chemokine system in diverse forms of macrophage activation and polarization. AB - Plasticity and functional polarization are hallmarks of the mononuclear phagocyte system. Here we review emerging key properties of different forms of macrophage activation and polarization (M1, M2a, M2b, M2c), which represent extremes of a continuum. In particular, recent evidence suggests that differential modulation of the chemokine system integrates polarized macrophages in pathways of resistance to, or promotion of, microbial pathogens and tumors, or immunoregulation, tissue repair and remodeling. PMID- 15530840 TI - TLRs: Professor Mechnikov, sit on your hat. AB - Toll-like receptors (TLRs) are sensors of foreign microbial products, which initiate host defense responses in all multicellular organisms examined to date. They are the target for most adjuvants, are essential for the establishment of memory in T and B cells and provoke inflammation. They program dendritic cells in their interaction with Th1 cells and their signalling pathways enable a tailoring of host defense responses to the provoking microbe previously unsuspected in the innate arm of immunity. Their discovery and characterisation fills a void in immunology and is the culmination of an effort that began with one of immunology's founding fathers, Elie Mechnikov. Targeting TLRs therapeutically now has the potential to impact on how we treat infectious and inflammatory diseases. PMID- 15530841 TI - Sex, MHC and complement C4 in autoimmune diseases. AB - Autoimmune diseases are estimated to affect 10-50 million people in the United States, and untold millions worldwide. Nearly 80% of all people with autoimmune diseases are women, and a strong association of these diseases with MHC genes has been known for some time. However, very little is known about what causes autoimmune diseases or the factors that lead to disease recurrence. The sex associated differences in multiple sclerosis (MS) and the mouse model of MS, experimental autoimmune encephalomyelitis (EAE), are associated with MHC genetic background, sex hormone levels and cytokine production. The implication of these factors has aided the identification of new autosomal genetic susceptibility loci. Complete deficiencies of early complement components are strongly associated with systemic lupus erythematosus (SLE) but the role of complement proteins in SLE is not yet clear. Recent data suggest that quantitative and qualitative diversities of the MHC-linked complement C4 among different ethnic groups can be important in the susceptibility and disease severity of SLE. PMID- 15530842 TI - Physical methods for structure, dynamics and binding in immunological research. AB - We present four experimental physical methods--X-ray and neutron diffraction, nuclear magnetic resonance spectroscopy, mass spectrometry and calorimetry--and two computational methods--molecular dynamics simulations and electrostatics calculations--which are general and widely applicable in the study of protein structure, dynamics and binding. These methods are useful tools for biologists that lead to structure-function, dynamics-function and binding-function correlations, in efforts to understand biomolecular function. Standard and emerging technologies within these methods are discussed and representative examples of applications in immunology are presented, from antigen-antibody, complement and MHC-T-cell receptor research. The examples demonstrate the power of the reviewed methods in immunological studies at the molecular level. PMID- 15530843 TI - Implicit alcohol-related cognitions in a clinical sample of heavy drinkers. AB - Patients undergoing treatment in an alcohol-rehabilitation clinic performed an Extrinsic Affective Simon Task (EAST) and two Implicit Association Tests (IATs) that were designed to measure implicit attitudes toward alcohol or implicit alcohol-arousal associations. The EAST and IAT data indicated that patients had a more negative implicit attitude toward alcohol than toward softdrinks and possessed strong implicit alcohol-arousal associations. Our results replicate and extend those of Wiers et al. (J. Abnormal Psychol 111 (2002) 648). PMID- 15530844 TI - Efficacy of cognitive therapy for depression among women with metastatic cancer: a single-case experimental study. AB - Although depression is frequent among patients with advanced cancer, very few studies have been conducted on its treatment. The objective of this study was to evaluate the efficacy of cognitive therapy for depression in women with metastatic cancer, using a multiple baseline experimental design. Six participants were enrolled in the study and were asked to complete daily and weekly mood assessments. Intervention time-series analyses conducted on daily mood data showed statistically significant improvement of depression symptoms, more importantly anhedonia, and associated features (i.e., anxiety, fatigue) for each participant. These improvements were also found to be clinically significant at post-treatment. PMID- 15530845 TI - Metacognitive therapy for PTSD: a preliminary investigation of a new brief treatment. AB - The effectiveness of a new treatment for post traumatic stress disorder (PTSD) is addressed. Treatment was based on a metacognitive theory of mechanisms by which natural traumatic processing is enabled or hindered by coping strategies. It suggests that elimination of worry/rumination, of maladaptive attention strategies, and enhancing metacognitive flexibility, will permit natural processing and a return to normal cognition. An A-B direct replication series (n = 6) with follow-up assessments at 3, 6, and 18-41 months was implemented. Treatment commenced 3-10 months post-trauma. All patients showed large and statistically significant improvements in general emotion and specific PTSD measures. Gains were maintained at follow-up. Two further consecutively referred patients were treated at 8 and 12 months post-trauma to add to sample size (n = 8). Overall Post treatment effect sizes were large, ranging from 3.0 to 5.0. Further evaluations are clearly warranted. PMID- 15530846 TI - An approach to psychotherapy toleration: the Distress/Endorsement Validation Scale (DEVS) for clinical outcome studies. AB - The issue of treatment tolerance within the field of psychotherapy is, at best, a nebulous construct and has been commonly evaluated via rates of subject attrition and homework compliance. This research presents the psychometric properties of a ten-item scale which endeavours to measure treatment distress and participant endorsement of therapy protocols used in clinical research. Two factors emerged and the subscales of Distress and Endorsement were derived. These subscales displayed good reliability with acceptable inter-item correlations within each subscale. The subscales were also able to differentiate the perspectives of male Vietnam veterans from their spouses on a lifestyle management course at the termination of intervention. However, this scale also displayed a cognitive behavioural trauma treatment protocol and eye movement desensitisation and reprocessing to be equivalent in treatment distress and participant endorsement in the treatment of post-traumatic stress disorder. Preliminary findings suggest that the relationship between these two subscales and outcome may, to some extent, be population specific. First evidence suggests that intervention distress ratings may be influenced by severity of presentation, whilst endorsement ratings are more influenced by symptomatic improvement over time. Suggestions for future research are presented and the full questionnaire is attached as an appendix. PMID- 15530847 TI - The significance of focal myoepithelial cell layer disruptions in human breast tumor invasion: a paradigm shift from the "protease-centered" hypothesis. AB - Human breast epithelium and the stroma are separated by a layer of myoepithelial (ME) cells and basement membrane, whose disruption is a prerequisite for tumor invasion. The dissolution of the basement membrane is traditionally attributed primarily to an over-production of proteolytic enzymes by the tumor or the surrounding stromal cells. The results from matrix metalloproteinase inhibitor clinical trials, however, suggest that this "protease-centered" hypothesis is inadequate to completely reflect the molecular mechanisms of tumor invasion. The causes and signs of ME cell layer disruption are currently under-explored. Our studies revealed that a subset of pre- and micro-invasive tumors contained focal disruptions in the ME cell layers. These disruptions were associated with immunohistochemical and genetic alterations in the overlying tumor cells, including the loss of estrogen receptor expression, a higher frequency of loss of heterozygosity, and a higher expression of cell cycle, angiogenesis, and invasion related genes. Focal ME layer disruptions were also associated with a higher rate of epithelial proliferation and leukocyte infiltration. We propose the novel hypothesis that a localized death of ME cells and immunoreactions that accompany an external environmental insult or internal genetic alterations are triggering factors for ME layer disruptions, basement membrane degradation, and subsequent tumor progression and invasion. PMID- 15530848 TI - Reactive oxygen species mediate RANK signaling in osteoclasts. AB - RANKL, a member of tumor necrosis factor (TNF) superfamily, regulates the differentiation, activation, and survival of osteoclasts through binding to its cognate receptor, RANK. RANK can interact with several TNF-receptor-associated factors (TRAFs) and activates signaling molecules including Akt, NF-kappaB, and MAPKs. Although the transient elevation of reactive oxygen species (ROS) by receptor activation has been shown to act as a cellular secondary messenger, the involvement of ROS in RANK signaling pathways has been not characterized. In this study, we found that RANKL stimulated ROS generation in osteoclasts. Pretreatment of osteoclasts with the antioxidants N-acetyl-l-cystein and glutathione reduced RANKL-induced Akt, NF-kappaB, and ERK activation. The reduced NF-kappaB activity by antioxidants was associated with decreased IKK activity and IkappaBalpha phosphorylation. In contrast, antioxidants did not prevent TNF-alpha-induced Akt and NF-kappaB activation. Pretreatment with antioxidants also significantly reduced RANKL-induced actin ring formation, required for bone resorbing activity, and osteoclast survival. Taken together, our results suggest that ROS act as mediators in RANKL-induced signaling pathways and cellular events. PMID- 15530849 TI - Involvement of JAK2 upstream of the PI 3-kinase in cell-cell adhesion regulation by gastrin. AB - The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway has been implicated in cell transformation and proliferation. Besides aberrant cell proliferation, loss of cell-cell adhesion during epithelial mesenchymal transition (EMT) is an important event which occurs during development of epithelial cancers. However, the role of JAK-dependent pathways in this process is not known. We analyzed the involvement of these pathways in the regulation of E-cadherin-dependent cell-cell adhesion by gastrin, a mitogenic factor for gastrointestinal (GI) tract. We identified JAK2/STAT3 as a new pathway in gastrin signaling. We demonstrated that JAK2 functions as an upstream mediator of the phosphatidylinositol 3 (PI 3)-kinase activity in gastrin signaling. Indeed, we observed a coprecipitation of both kinases and an inhibition of gastrin-induced PI 3-kinase activation when JAK2 activity is blocked. We also demonstrated that loss of cell-cell adhesion and the increase in cell motility induced by gastrin required the activation of JAK2 and the PI 3-kinase. Indeed, the modifications in localization of adherens junctions proteins and the migration, observed in gastrin-stimulated cells, were reversed by inhibition of both kinases. These results described the involvement of JAK2 in the modulation of cell-cell adhesion in epithelial cells. They support a possible role of JAK2 in the epithelial-mesenchymal transition which occurs during malignant development. PMID- 15530850 TI - The von Hippel-Lindau tumor suppressor protein influences microtubule dynamics at the cell periphery. AB - The von Hippel-Lindau (VHL) protein protects microtubules (MTs) from destabilization by nocodazole treatment. Based on this fixed-cell assay with static end points, VHL has been reported to directly stabilize the MT cytoskeleton. To investigate the dynamic changes in MTs induced by VHL in living cells, we measured the influence of VHL on tubulin turnover using fluorescence recovery after photobleaching (FRAP). To this end, we engineered VHL-deficient renal cell carcinoma cells to constitutively incorporate fluorescently labeled tubulin and to inducibly express VHL. Induction of VHL in these cells resulted in a decrease of tubulin turnover as measured by FRAP at the cell periphery, while minimally influencing MT dynamics around the centrosome. Our data indicates that VHL changes the behavior of MTs dependent on their subcellular localization implying a role for VHL in cellular processes such as migration, polarization, and cell-cell interactions. Here we propose a complementary method to directly measure VHL-induced subcellular changes in microtubule dynamics, which may serve as a tool to study the effect of MT binding proteins such as VHL. PMID- 15530852 TI - TIMP-1 inhibits microvascular endothelial cell migration by MMP-dependent and MMP independent mechanisms. AB - It was reported over a decade ago that tissue inhibitor of metalloproteinases-1 (TIMP-1) suppresses angiogenesis in experimental models but the mechanism is still incompletely understood. This in vitro study focused on the molecular basis of TIMP-1-mediated inhibition of endothelial cell (EC) migration, a key step in the angiogenic process. Both recombinant human TIMP-1 and the synthetic MMP inhibitors, GM6001 and MMP-2-MMP-9 Inhibitor III, suppressed migration of human dermal microvascular endothelial cells (HDMVEC) in a dose-dependent fashion. The MMP-dependent inhibition of migration was associated with increased expression of the junctional adhesion proteins, VE-cadherin and PECAM-1, and VE-cadherin accumulation at cell-cell junctions. TIMP-1 also caused MMP-independent dephosphorylation of focal adhesion kinase (FAK) (pY397) and paxillin, which was associated with reduced number of F-actin stress fibers and focal adhesions. Moreover, TIMP-1 stimulated expression of PTEN that has been shown to reduce phosphorylation of FAK and inhibit cell migration. Our data suggest that TIMP-1 inhibits HDMVEC migration through MMP-dependent stimulation of VE-cadherin and MMP-independent stimulation of PTEN with subsequent dephosphorylation of FAK and cytoskeletal remodeling. PMID- 15530851 TI - Retinoids arrest breast cancer cell proliferation: retinoic acid selectively reduces the duration of receptor tyrosine kinase signaling. AB - Retinoic acid (RA) induces cell cycle arrest of hormone-dependent human breast cancer (HBC) cells. Previously, we demonstrated that RA-induced growth arrest of T-47D HBC cells required the activity of the RA-induced protein kinase, protein kinase Calpha (PKCalpha) [J. Cell Physiol. 172 (1997) 306]. Here, we demonstrate that RA treatment of T-47D cells interfered with growth factor signaling to downstream, cytoplasmic and nuclear targets. RA treatment did not inhibit epidermal growth factor (EGF) receptor activation but resulted in rapid inactivation. The lack of sustained EGFR activation was associated with transient rather than sustained association of the EGFR with the Shc adaptor proteins and activation of Erk 1/2 and with compromised induction of expression of immediate early response genes. Inhibiting the activity of PKCalpha, a retinoic acid induced target gene, prevented the effects of RA on cell proliferation and EGF signaling. Constitutive expression of PKCalpha, in the absence of RA, decreased cell proliferation and decreased EGF signaling. RA treatment increased steady state levels of the protein tyrosine phosphatase PTP-1C and all measured effects of RA on EGF receptor function were reversed by the tyrosine phosphate inhibitor orthovanadate. These results indicate that RA-induced target genes, particularly PKCalpha, prevent sustained growth factor signaling, uncoupling activated receptor tyrosine kinases and nuclear targets that are required for cell cycle progression. PMID- 15530853 TI - Dual mode regulation of migration by lysophosphatidic acid in human gastric cancer cells. AB - Lysophosphatidic acid (LPA), which interacts with at least three G protein coupled receptors (GPCRs), LPA1/Edg-2, LPA2/Edg-4, and LPA3/Edg-7, is a lipid mediator with diverse effects on various cells. Here, we investigated the expression profiles of LPA receptors and patterns of LPA-induced migration in gastric cancer cells. Northern blot analysis revealed that various gastric cancer cells expressed variable levels of LPA1, LPA2, and LPA3 without a consistent pattern. Using a Boyden chamber assay, LPA markedly increased cell migration of LPA1-expressing cells, the effects of which were almost totally abrogated by Ki16425, an LPA antagonist against LPA1 and LPA3. In contrast, LPA by itself did not significantly induce migration in MKN28 and MKN74 cells, which exclusively expressed LPA2. However, when hepatocyte growth factor (HGF) was placed with LPA in the lower chamber, LPA induced migration of these cells in a dose-dependent manner. Immunoprecipitation analysis revealed that LPA induced transient tyrosine phosphorylation of c-Met in LPA2-expressing cells, which suggests that the transactivation of c-Met by LPA causes a cooperative migratory response with HGF to these cells. Our results indicate that LPA regulates the migration of gastric cancer cells in a receptor-specific manner and suggest that the expression pattern of LPA receptors may affect the metastatic behavior of gastric cancer. PMID- 15530854 TI - Adhesion-mediated signal transduction in human articular chondrocytes: the influence of biomaterial chemistry and tenascin-C. AB - Chondrocyte 'dedifferentiation' involves the switching of the cell phenotype to one that no longer secretes extracellular matrix found in normal cartilage and occurs frequently during chondrocyte expansion in culture. It is also characterized by the differential expression of receptors and intracellular proteins that are involved in signal transduction pathways, including those associated with cell shape and actin microfilament organization. The objective of this study was to examine the modulation of chondrocyte phenotype by cultivation on polymer substrates containing poly(ethylene glycol) (PEG). We observed differential arrangement of actin organization in articular chondrocytes, depending on PEG length. When cultivated on 300 g/mol PEG substrates at day 19, chondrocytes had lost intracellular markers characteristic of the differentiated phenotype, including type II collagen and protein kinase C (PKC). On these surfaces, chondrocytes also expressed focal adhesion and signaling proteins indicative of cell attachment, spreading, and FA turnover, including RhoA, focal adhesion kinase, and vinculin. The switch to a dedifferentiated chondrocyte phenotype correlated with integrin expression. Conversely, the expression of CD44 receptors coincided with chondrogenic characteristics, suggesting that binding via these receptors could play a role in maintaining the differentiated phenotype on such substrates. These effects can be similar to those of compounds that interfere in intracellular signaling pathways and can be utilized to engineer cellular response. PMID- 15530855 TI - Signaling pathway requirements for induction of senescence by telomere homolog oligonucleotides. AB - Cellular senescence is a major defense against cancer. In human fibroblasts, suppressing both the p53 and pRb pathways is necessary to bypass replicative senescence as well as senescence induced by ectopic expression of a dominant negative form of the telomere repeat binding factor 2, TRF2(DN). We recently reported that exposure to oligonucleotides homologous to the telomere 3' overhang (T-oligos) activates both the p53 and pRb pathways and leads to senescence in primary human fibroblasts. To further characterize T-oligo-induced senescence, we compared established isogenic fibroblast cell lines lacking functional p53 and/or pRb pathways to the normal parental line. Here, we report that, as in physiologic senescence, inactivation of both the p53 and pRb pathways is necessary to suppress T-oligo-induced senescence. Moreover, T-oligo rapidly induces senescence in a malignant fibroblast-derived cell line, demonstrating the potential of using T-oligo as a novel anticancer therapeutic. Our data support the hypothesis that exposure of the TTAGGG tandem repeat telomere 3' overhang sequence is the event that initiates signaling through DNA damage response pathways after experimental telomere disruption, serial passage, or acute genomic damage of normal cells. PMID- 15530856 TI - Stem cell factor is a chemoattractant and a survival factor for CNS stem cells. AB - Migration of neural cells to their final positions is crucial for the correct formation of the central nervous system. Several extrinsic factors are known to be involved in the regulation of neural migration. We asked if stem cell factor (SCF), well known as a chemoattractant and survival factor in the hematopoietic lineage, could elicit similar responses in neural stem cells. For that purpose, a microchemotaxis assay was used to study the effect of SCF on migration of neural stem cells from the embryonic rat cortex. Our results show that SCF-induced chemotaxis and that specific antibodies to SCF or tyrosine kinase inhibitors abolished the migratory response. The SCF-receptor, Kit, was expressed in neural stem cells and in their differentiated progeny. We also show that SCF is a survival factor, but not a mitogen or a differentiation factor for neural stem cells. These data suggest a role for SCF in cell migration and survival in the developing cortex. PMID- 15530858 TI - Poly(lactide-co-glycolide) nanoparticles as a carrier system for delivering cysteine protease inhibitor cystatin into tumor cells. AB - Cystatins are able to inhibit the tumor-associated activity of intracellular cysteine proteases cathepsins B and L and have been suggested as potential anticancer drugs. We have incorporated chicken cystatin, a model protein inhibitor of cysteine proteases, in poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) to improve its bioavailability and delivery into tumor cells. Cystatin-loaded NPs, 300-350 nm in diameter, were prepared by the double emulsion solvent diffusion method using low energy emulsification to preserve the biological activity of the protein. PLGA NPs and cystatin-loaded PLGA NPs at concentrations higher than 80 microg/ml were cytotoxic towards MCF-10A neoT cells, but not free cystatin at concentrations up to 5 microM. To visualize the uptake of cystatin into living MCF-10A neoT cells, NPs loaded with Alexa Fluor 488-labeled cystatin were added to the culture medium. They rapidly internalized into the cells, whereas the uptake of free-labeled cystatin was very slow. Cystatin, released from the NPs, effectively inhibited cathepsin B activity, as detected by degradation of specific Z-Arg-Arg cresyl violet substrate. In contrast, the same amount of free cystatin showed no inhibition of intracellular cathepsin B. Our results show that PLGA NPs are a useful carrier system for rapid delivery of protein inhibitors into tumor cells, enabling effective inhibition of intracellular proteolysis. The approach can be applied to other protein drugs active against intracellular targets. PMID- 15530859 TI - Endothelial cells within embryonic skeletal muscles: a potential source of myogenic progenitors. AB - We investigated whether the vessel-associated or endothelial cells within mouse embryo muscles can be a source of myogenic progenitors. Immunodetection of the stem cell surface markers, CD34 and Flk1, which are known to characterize the endothelial lineage, was done throughout the course of embryo muscle development. Both markers appeared to be restricted to the vessel-associated cells. On the basis of CD34 labeling, the reactive cells were purified by magnetic-bead selection from the limb muscles of 17-dpc desmin+/-LacZ mouse embryos and characterized by fluorescence-activated cell sorting. The cells in the selected CD34(+) population appeared to be approximately 95% positive for Flk1, but usually negative for CD45. We demonstrated that in vitro the CD34(+)/Flk1(+) population differentiated into endothelial cells and skeletal myofibers. When transplanted into mdx mouse muscle, this population displayed a high propensity to disperse within the recipient muscle, fuse with the host myofibers, and restore dystrophin expression. The marked ability of the embryonic muscle endothelial cells to activate myogenic program could be related to their somitic origin. PMID- 15530857 TI - An activating mutant of Cdc42 that fails to interact with Rho GDP-dissociation inhibitor localizes to the plasma membrane and mediates actin reorganization. AB - Cdc42 is a member of the Rho family of GTPases and plays an important role in the regulation of actin cytoskeletal organization. Activation of Cdc42 and associated signal transduction cascades are dependent upon proper localization of this GTPase. The studies described herein address the hypothesis that Rho GDP dissociation inhibitor, RhoGDI, plays an essential role in the translocation of Cdc42 to signaling complexes at the plasma membrane and is essential for Cdc42 mediated actin cytoskeletal rearrangements. An activating mutant of Cdc42 that is RhoGDI-binding defective (Cdc42(G12V/R66E)) is characterized and used as a tool to study the functional importance of the Cdc42-RhoGDI interaction. Overexpression of mycCdc42(G12V/R66E) in COS-7 cells results in actin cytoskeletal rearrangements that are indistinguishable from those stimulated by overexpression of mycCdc42(G12V). In addition, the G12V activating mutant of Cdc42 was overexpressed in mesangial cells that are null for RhoGDI expression. MycCdc42(G12V) stimulation of filopodia formation in these cells was indistinguishable from that observed in wild-type mesangial cells. Taken together, the results presented herein indicate that although RhoGDI is a critical regulator of guanine nucleotide binding, cycling of Cdc42 between membranes and the cytosol and cellular transformation, it is not essential for Cdc42-mediated organization of the actin cytoskeleton. PMID- 15530860 TI - Identification and molecular cloning of a functional GDP-fucose transporter in Drosophila melanogaster. AB - Nucleotide sugar transporters play a central role in the process of glycosylation. They are responsible for the translocation of nucleotide sugars from the cytosol, their site of synthesis, into the Golgi apparatus where the activated sugars serve as substrates for a variety of glycosyltransferases. We and others have recently identified and cloned the first GDP-fucose transporters of H. sapiens and C. elegans. Based on sequence similarity, we could identify a putative homolog in Drosophila melanogaster showing about 45% identity on protein level. The gene (CG9620) encodes a highly hydrophobic, multi-transmembrane spanning protein of 38.1 kDa that is localized in the Golgi apparatus. In order to test whether this protein serves as a GDP-fucose transporter, we performed complementation studies with fibroblasts from a patient with LADII (leukocyte adhesion deficiency II) which exhibit a strong reduction of fucosylation due to a point mutation in the human GDP-fucose transporter gene. We show that transient transfection of these cells with the Drosophila CG9620 cDNA corrects the GDP fucose transport defect and reestablishes fucosylation. This study gives experimental proof that the product of the in silico identified Drosophila gene CG9620 serves as a functional GDP-fucose transporter. PMID- 15530861 TI - In vivo overexpression of tumstatin domains by tumor cells inhibits their invasive properties in a mouse melanoma model. AB - Our previous studies demonstrated that a synthetic peptide encompassing residues 185-203 of the noncollagenous (NC1) domain of the alpha3 chain of type IV collagen, named tumstatin, inhibits in vitro melanoma cell proliferation and migration. In the present study, B16F1 melanoma cells were stably transfected to overexpress the complete tumstatin domain (Tum 1-232) or its C-terminal part, encompassing residues 185-203 (Tum 183-232). Tumstatin domain overexpression inhibited B16F1 in vitro cell proliferation, anchorage-independent growth, and invasive properties. For studying the in vivo effect of overexpression, representative clones were subcutaneously injected into the left side of C57BL6 mice. In vivo tumor growth was decreased by -60% and -56%, respectively, with B16F1 cells overexpressing Tum 1-232 or Tum 183-232 compared to control cells. This inhibitory effect was associated with a decrease of in vivo cyclin D1 expression. We also demonstrated that the overexpression of Tum 1-232 or Tum 183 232 induced an in vivo down-regulation of proteolytic cascades involving matrix metalloproteinases (MMPs), especially the production or activation of MMP-2, MMP 9, MMP-13, as well as MMP-14. The plasminogen activation system was also altered in tumors with a decrease of urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA) and a strong increase of plasminogen activator inhibitor-1 (PAI-1). Collectively, our results demonstrate that tumstatin or its C-terminal antitumor fragment, Tum 183-232, inhibits in vivo melanoma progression by triggering an intracellular transduction pathway, which involves a cyclic AMP (cAMP)-dependent mechanism. PMID- 15530862 TI - Topology of genes and nontranscribed sequences in human interphase nuclei. AB - Knowledge about the functional impact of the topological organization of DNA sequences within interphase chromosome territories is still sparse. Of the few analyzed single copy genomic DNA sequences, the majority had been found to localize preferentially at the chromosome periphery or to loop out from chromosome territories. By means of dual-color fluorescence in situ hybridization (FISH), immunolabeling, confocal microscopy, and three-dimensional (3D) image analysis, we analyzed the intraterritorial and nuclear localization of 10 genomic fragments of different sequence classes in four different human cell types. The localization of three muscle-specific genes FLNA, NEB, and TTN, the oncogene BCL2, the tumor suppressor gene MADH4, and five putatively nontranscribed genomic sequences was predominantly in the periphery of the respective chromosome territories, independent from transcriptional status and from GC content. In interphase nuclei, the noncoding sequences were only rarely found associated with heterochromatic sites marked by the satellite III DNA D1Z1 or clusters of mammalian heterochromatin proteins (HP1alpha, HP1beta, HP1gamma). However, the nontranscribed sequences were found predominantly at the nuclear periphery or at the nucleoli, whereas genes tended to localize on chromosome surfaces exposed to the nuclear interior. PMID- 15530863 TI - High extracellular calcium attenuates adipogenesis in 3T3-L1 preadipocytes. AB - We studied the effect of extracellular Ca(2+) concentration ([Ca(2+)](e)) on adipocyte differentiation. Preadipocytes exposed to continuous [Ca(2+)](e) higher than 2.5 mmol/l accumulated little or no cytoplasmic lipid compared to controls in 1.8 mmol/l [Ca(2+)](e). Differentiation was monitored by Oil Red O staining of cytoplasmic lipid and triglyceride assay of accumulated lipid, by RT-PCR analysis of adipogenic markers, and by the activity of glycerol-3-phosphate dehydrogenase (GPDH). Elevated [Ca(2+)](e) inhibited expression of peroxisome proliferator activated receptor gamma, CCAAT/enhancer binding protein alpha, and steroid regulatory binding element protein. High [Ca(2+)](e) significantly inhibited differentiation marker expression including adipocyte fatty acid binding protein, and GPDH. The decrease in Pref-1 expression that accompanied differentiation also was prevented by high [Ca(2+)](e). Treatment of 3T3-L1 cells with high [Ca(2+)](e) did not significantly affect cell number or viability and did not trigger apoptosis. Levels of intracellular Ca(+2) remained unchanged in various [Ca(2+)](e). Treatment of 3T3-L1 with pertussis toxin (PTX) partially restored lipid accumulation and increased differentiation markers in cells treated with 5 mmol/l [Ca(2+)](e). 'Classical' parathyroid cell Ca(2+) sensing receptors (CaSR) were not detected either by RT-PCR or by Western blotting. These results suggest that continuous exposure to high [Ca(2+)](e) inhibits preadipocyte differentiation and that this may involve a G-protein-coupled mechanism mediated by a novel Ca(2+) sensor or receptor. PMID- 15530864 TI - Nm23-M2/NDP kinase B induces endogenous c-myc and nm23-M1/NDP kinase A overexpression in BAF3 cells. Both NDP kinases protect the cells from oxidative stress-induced death. AB - The nm23 gene family encodes nucleoside diphosphate kinases (NDPKs) which supply the cell with (d)NTPs. The human NDPKB, also known as the PuF protein, binds the c-myc promoter and transactivates the c-myc protooncogene. We have now studied the effects of mouse NDPKA and NDPKB overexpression on endogenous c-myc transactivation in the mouse BAF3 and the rat PC12 cell lines. c-myc transcripts were found to be up-regulated by NDPKB only in the BAF3 line. This suggests that c-myc transcriptional control via NDPKB depends on the presence of cell-specific co-factors. Unexpectedly, NDPKB also induced NDPKA expression. This new effect was found in both cell lines, suggesting that NDPKB-dependent nm23-M1 gene transactivation requires cis and/or trans elements different from those involved in c-myc transactivation. Moreover, the BAF3 cell proliferation capacities were found to be independent of NDPKA or B cell contents. Interestingly, cell death induced by c-myc overexpression or H(2)O(2) exposure was decreased in nm23 transfected compared to control BAF3 cells. These data collectively suggest that NDPKs might improve cell survival by a mechanism coupling DNA repair and transcriptional regulation of genes involved in DNA damage response. PMID- 15530865 TI - HMG-CoA reductase inhibitors induce COX-2 gene expression in murine macrophages: role of MAPK cascades and promoter elements for CREB and C/EBPbeta. AB - Except functioning as lipid-lowering agents, HMG-CoA inhibitors, statins, are good tools to clarify the signaling role of small G proteins. In this study, we found in murine RAW264.7 macrophages, statins within 1-30 microM stimulated COX-2 gene transcription and PGE(2) formation, displaying potencies as lovastatin > fluvastatin > atorvastatin >> pravastatin. Transfection experiments with COX-2 promoter construct showed the necessity of C/EBPbeta and CRE promoter sites, but not NF-kappaB promoter site. Effects of statins on the activation of COX-2 promoter, induction of COX-2 protein, and PGE(2) production were all prevented by mevalonate and prenylated metabolites, FPP and GGPP. Consistent with the effect of statins, manumycin A, farnesyltransferase inhibitor, and geranylgeranyltransferase inhibitor increased PGE(2) production and COX-2 induction. Likewise, toxin B, an inhibitor of Rho family members, caused a prominent COX-2 induction. Results also indicated that tyrosine kinase, ERK, and p38 MAPK play essential roles in statin action. Taken together, these results not only demonstrate a unique action of statins in the upregulation of COX-2 expression in macrophages, but also suggest a negative role controlled by small G proteins in COX-2 gene regulation. Removal of this negative control by impairing G protein prenylation with statins leads to MAPKs activation and promotes COX-2 gene expression through the activation at CRE and C/EBPbeta sites. PMID- 15530866 TI - MIBG, an inhibitor of arginine-dependent mono(ADP-ribosyl)ation, prevents differentiation of L6 skeletal myoblasts by inhibiting expression of myogenin and p21(cip1). AB - The development of skeletal muscle is controlled by a highly synchronized series of cellular events, and various signals from both inside and outside the cell play a role in the switch from multipotential mesodermal stem cells to muscle fibers. Meta-iodobenzylguanidine (MIBG), an inhibitor of mono(ADP-ribosyl)ation, has been shown to prevent terminal differentiation of skeletal myoblasts; however, its mechanism of action has not been established. We recently reported that MIBG is capable of preventing phenotypic modulation of smooth muscle cells by interfering with specific trans-acting factors [L. Yau, B. Litchie, S. Thomas, B. Storie, N. Yurkova, P. Zahradka, Endogenous mono-ADP-ribosylation mediates smooth muscle cell proliferation and migration via protein kinase N-dependent induction of c-fos expression. Eur. J. Biochem. 270 (2003) 101-110.]. We therefore examined the effect of MIBG on select myogenic regulatory factors known to control terminal differentiation. It was confirmed that MIBG, but not inhibitors of poly-ADP-ribose polymerase (3-aminobenzamide, PD128763), inhibits fusion of L6 skeletal myoblasts in a concentration-dependent manner. Moreover, inhibition by MIBG correlated with a failure to induce expression of myogenin and p21(cip1), while levels of MyoD and MEF2 were unaffected. Time-of-addition studies revealed that MIBG also affected a late event possibly linked to cell fusion. Finally, arginine-dependent mono(ADP-ribosyl)transferase activity increased over the first 24 h of the differentiation period. These data support a role for arginine-dependent mono(ADP-ribosyl)transferase as an essential positive regulator of differentiation in skeletal muscle cells that operates by modulating the expression of specific myogenic factors. PMID- 15530867 TI - Wave2 activates serum response element via its VCA region and functions downstream of Rac. AB - WAVE2 is a member of the WASP/WAVE family of protein effectors of actin reorganization and cell movement. In this report, we demonstrate that WAVE2 overexpression induces serum response element (SRE) activation through serum response factor. A WAVE2 mutant lacking the VCA region did not induce SRE activation and actin polymerization. WAVE2-induced SRE activation was blocked by exposure of cells to Latrunculin A, or overexpression of actin mutant R62D. The DeltaVCA mutant inhibited Rac V12-induced SRE activation, suggesting that WAVE2 lies downstream of Rac. Similar deletion of the VCA domain of WASP attenuated Cdc42 V12-mediated SRE activation, suggesting that WAVE2 acts in relation to Rac as WASP acts in relation to Cdc42. WAVE2 overexpression did not activate NF kappaB. PMID- 15530868 TI - Motoneurons crave glial cell line-derived neurotrophic factor. AB - This is a commentary on the developmental and therapeutic relevance of recent studies in the glial fibrillary acid protein (GFAP)-glial cell line-derived neurotrophic factor (GDNF) transgenic mouse reported by Zhao et al. (2004). This interesting study demonstrated that increased expression of GDNF in astrocytes increases the number of neighboring motoneurons of certain motoneuron subpopulations by diminishing programmed cell death during development. In addition, astrocyte-derived GDNF was shown to protect facial motoneurons from injury-induced cell death. Since this is the first direct demonstration that secretion of GDNF from astrocytes in the CNS can affect motoneuron development in utero and motoneuron survival after axotomy, novel approaches for motor neuron disease are suggested. The known target neurons that respond to GDNF are reviewed, as are studies using GDNF gene delivery in animal models of amyotrophic lateral sclerosis (ALS). It is postulated that GDNF is a factor to which many motoneurons respond along their whole extent from soma to axon to terminal. PMID- 15530869 TI - CNS stem and progenitor cell differentiation into functional neuronal circuits in three-dimensional collagen gels. AB - The mammalian central nervous system (CNS) has little capacity for self-repair after injury, and neurons are not capable of proliferating. Therefore, neural tissue engineering that combines neural stem and progenitor cells and biologically derived polymer scaffolds may revolutionize the medical approach to the treatment of damaged CNS tissues. Neural stem and progenitor cells isolated from embryonic rat cortical or subcortical neuroepithelium were dispersed within type I collagen, and the cell-collagen constructs were cultured in serum-free medium containing basic fibroblast growth factor. The collagen-entrapped stem and progenitors actively expanded and efficiently generated neurons, which developed neuronal polarity, neurotransmitters, ion channels/receptors, and excitability. Ca2+ imaging showed that differentiation from BrdU+/TuJ1- to BrdU-/TuJ1+ cells was accompanied by a shift in expression of functional receptors for neurotransmitters from cholinergic and purinergic to predominantly GABAergic and glutamatergic. Spontaneous postsynaptic currents were recorded by patch-clamping from precursor cell-derived neurons and these currents were partially blocked by 10-microM bicuculline, and completely blocked by additional 10 microM of the kainate receptor antagonist CNQX, indicating an appearance of both GABAergic and glutamatergic synaptic activities. Staining with endocytotic marker FM1-43 demonstrated active synaptic vesicle recycling occurring among collagen-entrapped neurons. These results show that neural stem and progenitor cells cultured in 3D collagen gels recapitulate CNS stem cell development; this is the first demonstration of CNS stem and progenitor cell-derived functional synapse and neuronal network formation in a 3D matrix. The proliferative capacity and neuronal differentiating potential of neural progenitors in 3D collagen gels suggest their potential use in attempts to promote neuronal regeneration in vivo. PMID- 15530870 TI - Acute transplantation of glial-restricted precursor cells into spinal cord contusion injuries: survival, differentiation, and effects on lesion environment and axonal regeneration. AB - Transplantation of stem cells and immature cells has been reported to ameliorate tissue damage, induce axonal regeneration, and improve locomotion following spinal cord injury. However, unless these cells are pushed down a neuronal lineage, the majority of cells become glia, suggesting that the alterations observed may be potentially glially mediated. Transplantation of glial-restricted precursor (GRP) cells--a precursor cell population restricted to oligodendrocyte and astrocyte lineages--offers a novel way to examine the effects of glial cells on injury processes and repair. This study examines the survival and differentiation of GRP cells, and their ability to modulate the development of the lesion when transplanted immediately after a moderate contusion injury of the rat spinal cord. GRP cells isolated from a transgenic rat that ubiquitously expresses heat-stable human placental alkaline phosphatase (PLAP) were used to unambiguously detect transplanted GRP cells. Following transplantation, some GRP cells differentiated into oligodendrocytes and astrocytes, retaining their differentiation potential after injury. Transplanted GRP cells altered the lesion environment, reducing astrocytic scarring and the expression of inhibitory proteoglycans. Transplanted GRP cells did not induce long-distance regeneration from corticospinal tract (CST) and raphe-spinal axons when compared to control animals. However, GRP cell transplants did alter the morphology of CST axons toward that of growth cones, and CST fibers were found within GRP cell transplants, suggesting that GRP cells may be able to support axonal growth in vivo after injury. PMID- 15530871 TI - Modification of the regenerative response of dorsal column axons by olfactory ensheathing cells or peripheral axotomy in adult rat. AB - The regeneration of sciatic-dorsal column (DC) axons following DC crush injury and treatment with olfactory ensheathing cells (OECs) and/or sciatic axotomy ("conditioning lesion") was evaluated. Sciatic-DC axons were examined with a transganglionic tracer, cholera toxin conjugated to horseradish peroxidase, and evaluated at chronic time points, 2-26 weeks post-lesion. With DC injury alone (n = 7), sciatic-DC axons were localized to the caudal border of the lesion terminating in reactive end bulbs with no indication of growth into the lesion. In contrast, treatment with either a heterogeneous population of OECs (equal numbers of p75- and fibronectin-positive OECs) (n = 9) or an enriched population of OECs (75% p75-positive OECs) (n = 6) injected either directly into the lesion or 1-mm rostral and caudal to the injury, stimulated DC axon growth into the lesion. A similar regenerative response was observed with a conditioning lesion either concurrent to (n = 4) or 1 week before (n = 4) the DC injury. In either of the latter two paradigms, some DC axons grew across the injury, but no axons grew into the rostral intact spinal cord. Upon combining OEC treatment with the conditioning lesion (n = 21), the result was additive, increasing DC axon growth beyond the rostral border of the lesion in best cases. Additional factors that may limit DC regeneration were tested including formation of the glial scar (immunoreactivity to glial fibrillary acidic protein in astrocytes and to chondroitin sulfate proteoglycans), which remained similar between treated and untreated groups. PMID- 15530872 TI - Learning and memory impairment in rats with chronic atypical absence seizures. AB - Atypical absence seizures (AASs) represent a pediatric malignant seizure type that commonly exists as a component of Lennox-Gastaut syndrome. AAS involves both the hippocampal and thalamocortical circuitry in slow spike-and-wave discharges (SSWD) and is associated with cognitive dysfunction. The electrographic, behavioral, and pharmacological features of clinical AAS have been reproduced in rats chronically in the AY-9944 (AY) model. AY rats show spontaneous SSWD involving the hippocampus, a structure that is highly implicated in learning and memory. The purpose of the present study was to determine whether AY rats exhibit cognitive deficits that mirror those observed in AAS clinically. Hippocampal function was examined in AY animals both in vitro with electrophysiology (i.e., synaptic plasticity) and in vivo with a hippocampus-dependent radial arm maze (RAM) task that is designed to assess spatial cognition. In vitro tests of synaptic plasticity revealed impairments in long-term potentiation (LTP), paired pulse facilitation (PPF), and presynaptic depression (PD). Consistently, performance of AY animals in RAM revealed fewer perfect entries, a greater number of errors, and required more training days to learn the task than saline-treated controls. The abolishment of spontaneous seizures by ethosuximide failed to recover the perturbed spatial learning and working memory in AY animals. AY rats demonstrate altered hippocampal functioning as manifested by altered synaptic plasticity and cognition. The relationship between AAS and cognitive deficit remains uncertain and the pathophysiology of both in AY treated requires further investigation. PMID- 15530873 TI - Neurodegenerative alterations in the nigrostriatal system of trkB hypomorphic mice. AB - Brain-derived neurotrophic factor (BDNF) acts through the neurotrophin receptor TrkB and promotes survival and differentiation of dopaminergic ventral mesencephalic neurons. To further evaluate the role of TrkB in the nigrostriatal pathway, we studied neurotrophin levels, dopamine metabolism, and morphology of dopaminergic neurons of the substantia nigra (SN-DA) in young adult hypomorphic trkB mice (trkBfbz/fbz), which express only approximately 25% of wild type levels of TrkB. Tyrosine hydroxylase immunostaining revealed altered morphology of SN-DA neurons in trkBfbz/fbz when compared to wild type mice, in particular a significant enlargement of nuclear size. Cell counts revealed a pronounced loss of SN-DA neurons in these mice. Measurement of monoamine levels by high performance liquid chromatography (HPLC) showed that dopamine (DA) levels in the target field (striatum) were significantly elevated in trkBfbz/fbz compared to trkB+/fbz and wild type mice (P < 0.05), without altering DA turnover. Likewise, enzyme-linked immunosorbent assay (ELISA) for neurotrophic factors measurement showed that BDNF levels were increased in the striatum (P < 0.01) and frontal cortex (P < 0.005) of trkBfbz/fbz mice, but not in the SN when compared to trkB+/fbz and wild type mice. These data suggest that elevated neurotransmitter and neurotrophic factor levels might be a compensatory mechanism following dopaminergic cell loss in the SN. Thus, TrkB-activation seems essential for the maintenance of the nigrostriatal dopaminergic system. PMID- 15530874 TI - Effects of motor versus sensory nerve grafts on peripheral nerve regeneration. AB - Autologous nerve grafting is the current standard of care for nerve injuries resulting in a nerve gap. This treatment requires the use of sensory grafts to reconstruct motor defects, but the consequences of mismatches between graft and native nerve are unknown. Motor pathways have been shown to preferentially support motoneuron regeneration. Functional outcome of motor nerve reconstruction depends on the magnitude, rate, and precision of end organ reinnervation. This study examined the role of pathway type on regeneration across a mixed nerve defect. Thirty-six Lewis rats underwent tibial nerve transection and received isogeneic motor, sensory or mixed nerve grafts. Histomorphometry of the regenerating nerves at 3 weeks demonstrated robust nerve regeneration through both motor and mixed nerve grafts. In contrast, poor nerve regeneration was seen through sensory nerve grafts, with significantly decreased nerve fiber count, percent nerve, and nerve density when compared with mixed and motor groups (P < 0.05). These data suggest that use of motor or mixed nerve grafts, rather than sensory nerve grafts, will optimize regeneration across mixed nerve defects. PMID- 15530875 TI - Overexpression of glial cell line-derived neurotrophic factor in the CNS rescues motoneurons from programmed cell death and promotes their long-term survival following axotomy. AB - To study the role of one of the most potent motoneuron (MN) survival factors, glial cell line-derived neurotrophic factor (GDNF) derived from the CNS, we generated transgenic animals overexpressing GDNF under the control of an astrocyte-specific GFAP promoter. In situ hybridization revealed that GDNF was expressed at high levels in astrocytes throughout the brain and spinal cord. We analyzed the effects of CNS-derived GDNF on MN survival during the period of programmed cell death (PCD) and after nerve axotomy. In GFAP-GDNF mice at E15, E18, and P1, the survival of brachial MNs was increased on average by 30%, lumbar MNs by 20%, and thoracic MNs at P1 by 33%. GDNF also prevented MN PCD in several cranial motor nuclei. We demonstrated for the first time that the number of MNs in the mouse abducens nucleus was also increased by 40%, thus extending known MN populations that are responsive to GDNF. Next, we tested if GDNF could support complete and relatively long-term survival of MNs following neonatal facial nerve axotomy. We found that virtually all MNs (91%) in GFAP-GDNF mice survived for up to 18 weeks post-axotomy. This is the longest GDNF-mediated survival of neonatal MNs reported following axotomy. Most of surviving MNs were not atrophic, and MN specific ChAT and neurofilament immunoreactivity (IR) were preserved. Furthermore, GDNF attenuated axotomy-induced astroglial activation. These data demonstrate that overexpression of GDNF in the CNS has very profound effects on MN survival both during the PCD period and after neuronal injury. GFAP-GDNF mice will be valuable to study the effects of CNS-derived GDNF in mouse models of MN degenerative diseases and axonal regeneration in vivo. PMID- 15530876 TI - Rotenone potentiates dopamine neuron loss in animals exposed to lipopolysaccharide prenatally. AB - We previously demonstrated that treating gravid female rats with the bacteriotoxin lipopolysaccharide (LPS) led to the birth of offspring with fewer than normal dopamine (DA) neurons. This DA neuron loss was long-lived and associated with permanent increases in the pro-inflammatory cytokine tumor necrosis factor alpha (TNFalpha). Because of this pro-inflammatory state, we hypothesized that these animals would be more susceptible to subsequent exposure of DA neurotoxins. We tested this hypothesis by treating female Sprague-Dawley rats exposed to LPS or saline prenatally with a subtoxic dose of the DA neurotoxin rotenone (1.25 mg/kg per day) or vehicle for 14 days when they were 16 months old. After another 14 days, the animals were sacrificed. Tyrosine hydroxylase-immunoreactive (THir) cell counts were used as an index of DA neuron survival. Animals exposed to LPS prenatally or rotenone postnatally exhibited a 22% and 3%, respectively, decrease in THir cell counts relative to controls. The combined effects of prenatal LPS and postnatal rotenone exposure produced a synergistic 39% THir cell loss relative to controls. This loss was associated with decreased striatal DA and increased striatal DA activity ([HVA]/[DA]) and TNFalpha. Animals exposed to LPS prenatally exhibited a marked increase in the number of reactive microglia that was further increased by rotenone exposure. Prenatal LPS exposure also led to increased levels of oxidized proteins and the formation of alpha-Synuclein and eosin positive inclusions resembling Lewy bodies. These results suggest that exposure to low doses of an environmental neurotoxin like rotenone can produce synergistic DA neuron losses in animals with a preexisting pro-inflammatory state. This supports the notion that Parkinson's disease (PD) may be caused by multiple factors and the result of "multiple hits" from environmental toxins. PMID- 15530877 TI - In vivo upregulation of endogenous NGF in the rat brain by the alpha2 adrenoreceptor antagonist dexefaroxan: potential role in the protection of the basalocortical cholinergic system during neurodegeneration. AB - We have previously reported that the alpha2-adrenoceptor antagonist dexefaroxan protects against the degeneration of nucleus basalis magnocellularis (NbM) cholinergic neurons following cortical devascularization in the adult rat. Since nerve growth factor (NGF) is critical to the survival of NbM cholinergic neurons in the adult brain and its synthesis is known to be regulated by noradrenergic mechanisms, we examined whether the protective effect of dexefaroxan in the devascularization model was associated with regional induction of NGF biosynthesis. Dexefaroxan or vehicle was administered to rats via subcutaneous minipumps for 28 days following devascularization or sham operation procedures. In vehicle-treated devascularized rats, NGF protein levels in the cortex were increased at 5 days but had normalized by 2 weeks postoperation; NGF levels in NbM remained unchanged during this time. In dexefaroxan-treated devascularized rats, increases in NGF protein levels (2-fold) and immunoreactivity were maintained in both the cortex and NbM over the entire 28-day postoperation period; these increases were coincident with changes in functional markers characteristic of NGF's actions, including increases in choline acetyltransferase (ChAT), p75 and TrkA immunoreactivities, and a preservation of NbM cholinergic cell numbers. Dexefaroxan also increased NGF protein levels in sham-operated rats, but without any significant consequence to the otherwise normal NbM cholinergic phenotype in these animals. Results indicate that activation of endogenous NGF systems could contribute to the cholinergic protective effect of dexefaroxan in the cortical devascularization model, and provide further support for a potential therapeutic utility of dexefaroxan in neurodegenerative diseases where central cholinergic function is progressively compromised. PMID- 15530878 TI - Modification of the brain-derived neurotrophic factor gene: a portal to transform mesenchymal stem cells into advantageous engineering cells for neuroregeneration and neuroprotection. AB - Multipotential mesenchymal stem cells (MSCs) are ideal seed cells for recruiting the loss of neural cells due to their strong proliferative capacity, easy acquisition, and considerable tolerance of genetic modifications. After transduction of brain-derived neurotrophic factor (BDNF) gene via recombinant retroviral vectors into the human MSCs, nearly 100% of cells expressed BDNF (which were therefore transformed into BNDF-MSCs) as detected by immunocytochemistry, and the quantity of BDNF in the culture medium was increased by approximately 20,000-fold. In spite of the genomic integration of an exogenous gene, BDNF-MSCs did not present any structural aberration in the chromosomes. All trans-retinoic acid (RA) induction caused the BDNF-MSCs to differentiate into neural cells with significantly increased expressions of such neural-specific proteins as nestin, NeuN, O4, and glial fibrillary acidic protein (GFAP). The voltage-dependent K+/Ca2+ currents were recorded from the induced BDNF-MSCs using patch-clamp technique. Compared with the MSCs induced by both RA and BDNF, BDNF MSCs survived in significantly greater number in the induction medium, and also more cells were induced into neuron-like cells (NeuN, P < 0.01) and oligodendrocyte-like cells (O4, P < 0.05). We suppose that, once engrafted into human central nervous system, the BDNF-MSCs would not only recruit the neuronal losses, but also provide, by way of paracrine, large quantities of BDNF that effectively perform the functions of neuroprotection and neuroregeneration, promoting the activation of endogenous neural stem/progenitor cells and their chemotactic migration. On the other hand, the BDNF-MSCs that can survive in the host environment and differentiate subsequently into functional mature cells may also serve as specifically targeting vectors for ex vivo gene therapy. PMID- 15530879 TI - Motor neurons can preferentially reinnervate cutaneous pathways. AB - Previous work in the rat femoral nerve has shown that regenerating motor neurons preferentially reinnervate a terminal nerve branch to muscle as opposed to skin. This process has been termed preferential motor reinnervation (PMR) and has been interpreted as evidence that regenerating motor axons can differentiate between Schwann cell tubes that reside in muscle versus cutaneous terminal pathways. However, much of this previous work has been confounded by motor axons having access to target muscle during the regeneration period. The present experiments prevented muscle contact by regenerating motor axons. By 8 weeks under these conditions, significantly more motor neurons reinnervated the cutaneous pathway rather than the original muscle pathway. We propose that cutaneous and muscle terminal pathways are not inherently different in terms of their ability to support regeneration of motor neurons. Rather, we suggest that it is the relative level of trophic support provided by each nerve branch that determines whether motor axons will remain in that particular branch. Within the context of the femoral nerve model, our results suggest a hierarchy of trophic support for regenerating motor axons with muscle contact being the highest, followed by the length of the terminal nerve branch and/or contact with skin. PMID- 15530880 TI - Evidence that infiltrating neutrophils do not release reactive oxygen species in the site of spinal cord injury. AB - The release of reactive oxygen species (ROS) by neutrophils, which infiltrate the region of damage following spinal cord injury (SCI), was investigated to determine if such release is significant following spinal cord injury. The relationship of extracellular levels of hydroxyl radicals and hydrogen peroxide obtained by microdialysis sampling and oxidized protein levels in tissue to neutrophil infiltration following spinal cord injury was examined. Neither of the reactive oxygen species were elevated in the site of spinal cord injury relative to their concentrations in normal tissue at a time (24 h) when the numbers of neutrophils were maximum in the site of injury. Surprisingly, ablation with a neutrophil antiserum actually increased the level of oxidized proteins in Western blots. Thus, our findings are (1) that neutrophils, which infiltrate the site of damage following a spinal cord injury, do not release detectable quantities of reactive oxygen species; and (2) that the presence of neutrophils reduces the concentrations of oxidized proteins in the site of spinal cord injury. Therefore, release of reactive oxygen species by neutrophils does not contribute significantly to secondary damage following spinal cord injury. Reduced levels of oxidized proteins in the presence of neutrophils may reflect removal of damaged tissue by neutrophils. PMID- 15530881 TI - Functional significance of the ipsilateral hemisphere during movement of the affected hand after stroke. AB - Previous fMRI observations have suggested increased task-related activation of the ipsilateral cerebral motor cortex in patients recovering from stroke. This is generally taken to infer an increased output from this area, although the functional relevance of this has been questioned. Here, we use directed EEG coherence to reveal whether there is increased informational flow from the ipsilateral motor cortex following motor stroke, and through correlation with degree of recovery, establish that this pattern of activity is associated with limited functional improvement. Unrecovered (n = 14), recovered (n = 11) patients and healthy subjects (n = 16) performed an isometric grip task with either hand that corresponded to 25% of individual maximum force, while EEG was recorded. For unrecovered stroke patients, most task-related information flow between the sensorimotor cortices in the low beta band of the EEG came from the ipsilateral (undamaged) hemisphere during grip with the affected hand. This was not the case when they gripped with their unaffected hand, when cortical activity was driven from the contralateral sensorimotor cortex. The latter pattern was also seen in recovered patients and controls. These findings suggest a functional role for the ipsilateral hemisphere in organizing movement of the impaired limb following stroke, but only in those patients that do not make a good functional recovery. Patients making a fuller recovery organize movement-related cortical activity from the hemisphere contralateral to movement. PMID- 15530882 TI - Unilateral ischemic sensorimotor cortical damage in female rats: forelimb behavioral effects and dendritic structural plasticity in the contralateral homotopic cortex. AB - Previous studies in male rats with unilateral sensorimotor cortical (SMC) damage have demonstrated dendritic structural plasticity in the contralateral homotopic cortex and an enhancement of skilled reaching performance in the forelimb ipsilateral to the lesion compared to sham-operated rats. The purpose of this study was to determine if these findings could be replicated in an ischemic lesion model in female rats. Female rats were given sham operations or unilateral ischemic (endothelin-1 induced) damage in the forelimb representation area of the SMC opposite their preferred forelimb. Animals then received either 20 consecutive days of training on a skilled reaching task with the non preferred/unimpaired forelimb or no-training control procedures. The surface density of dendrites immunoreactive (IR) for microtubule-associated protein 2 (MAP2) was then measured in the motor cortex opposite the trained limb and/or lesion. Female rats with sufficiently large, but not very small, lesions performed better with the unimpaired forelimb than sham-operated rats on the reaching task. The post-lesion reaching performance was not found to be significantly dependent upon estrous stage at the time of surgery, in agreement with previous studies that failed to find sex or sex-hormone effects after other types of SMC damage. Additionally, there were major laminar-dependent increases in the surface density of MAP2 IR dendrites in the cortex opposite lesions and trained limbs. These findings in female rats are consistent with the dendritic and behavioral changes previously found in male rats. They extend these previous findings by indicating that lesion size is an important variable in the enhancement of reaching performance. PMID- 15530883 TI - Cytokines, nitric oxide, and cGMP modulate the permeability of an in vitro model of the human blood-brain barrier. AB - The endothelial cells (EC) of the microvasculature in the brain form the anatomical basis of the blood-brain barrier (BBB). In the present study, the effects of agents that modify the permeability of a well-established in vitro model of the human BBB were studied. The monolayers formed by confluent human brain microvessel endothelial cell (HBMEC) cultures are impermeable to the macromolecule tracer horseradish peroxidase (HRP) and have high electrical resistance. Exposure of HBMEC to various cytokines including TNF-alpha, IL-1beta, interferon gamma (IFN-gamma), or lipopolysaccharide (LPS) decreased transendothelial electrical resistance (TEER) mainly by increasing the permeability of the tight junctions. Primary cultures of HBMEC express endothelial nitric oxide synthase (eNOS) and produce low levels of NO. Treatment with the NO donors sodium nitroprusside (SNP) and DETA NONOate or the cGMP agonist 8-Br-cGMP significantly increased monolayer resistance. Conversely, inhibition of soluble guanylyl cyclase with ODQ rapidly decreased the resistance, and pretreatment of HBMEC with Rp-8-CPT-cGMPS, an inhibitor of cGMP-dependent protein kinase, partially prevented the 8-Br-cGMP-induced increase in resistance. Furthermore, NO donors and 8-Br-cGMP could also reverse the increased permeability of the monolayers induced by IL-1beta, IFN-gamma, and LPS. These results indicate that NO can decrease the permeability of the human BBB through a mechanism at least partly dependent on cGMP production and cGMP-dependent protein kinase activation. PMID- 15530884 TI - Astrocytosis, microglia activation, oligodendrocyte degeneration, and pyknosis following acute spinal cord injury. AB - Glial activation and degeneration are important outcomes in the pathophysiology of acute brain and spinal cord injury (SCI). Our main goal was to investigate the pattern of glial activation and degeneration during secondary degeneration in both gray matter (GM) and white matter (WM) following SCI. Adult rats were deeply anesthetized and injected with 20 nmol of N-methyl-D-aspartate (NMDA) into the ventral horn of rat spinal cord (SC) on T7. Animals were perfused after survival times of 1, 3, and 7 days. Ten-micrometer sections were submitted to immunocytochemistry for activated macrophages/microglia, astrocytes, oligodendrocytes, and myelin. Astrocyte activation was more intense in the vacuolated white matter than in gray matter and was first noticed in this former region. Microglial activation was more intense in the gray matter and was clear by 24 h following NMDA injection. Both astrocytosis and microglial activation were more intense in the later survival times. Conspicuous WM vacuolation was present mainly at the 3-day survival time and decreased by 7 days after the primary damage. Quantitative analysis revealed an increase in the number of pyknotic bodies mainly at the 7-day survival time in both ventral and lateral white matter. These pyknotic bodies were frequently found inside white matter vacuoles like for degenerating oligodendrocytes. These results suggest a differential pattern of astrocytosis and microglia activation for white and gray matter following SCI. This phenomenon can be related to the different pathological outcomes for this two SC regions following acute injury. PMID- 15530885 TI - Estrogen attenuates the MPTP-induced loss of dopamine neurons from the mouse SNc despite a lack of estrogen receptors (ERalpha and ERbeta). AB - Estrogen attenuates the loss of dopamine from striatum and dopamine neurons from the substantia nigra (SNc) in animal models of Parkinson's disease. Interestingly, estrogen receptors (ERalpha and ERbeta) are thought to be sparse or absent in mouse striatum and SNc. Since ERalpha is markedly induced in rodent cortex after ischemic injury, the present studies evaluated changes in ERs after acute treatment with the dopamine neurotoxin MPTP. Mice were injected daily with estradiol, injected with MPTP on day 6, and brains collected on day 9 or 13. Immunocytochemistry was then used to assess tyrosine hydroxylase (TH) in striatum and investigate the localization of ERalpha and ERbeta in the striatum and SNc. In addition, cryostat sections were hybridized with a riboprobe complementary to ERalpha or ERbeta mRNA. Evaluation of TH immunoreactivity revealed a dense network of fibers in the striatum of vehicle-treated animals, while a near complete loss of terminals was seen after MPTP treatment. When, however, mice were pretreated with estradiol, the MPTP-induced loss of TH was attenuated. Evaluation of ERalpha and ERbeta in the SNc and striatum demonstrated a sparse localization of both ERs in vehicle-treated mice, a pattern that did not change in animals treated with vehicle/MPTP or estradiol/MPTP. These data demonstrate that ERs are sparse in the mouse striatum and SNc and show that this pattern does not change after MPTP intoxication. This observation and the finding that estrogen affords some protection against MPTP suggest that estrogen may act via nuclear receptor independent mechanisms to protect dopamine neurons from toxins such as MPTP. PMID- 15530886 TI - Spontaneous regression of experimental gliomas--an immunohistochemical and MRI study of the C6 glioma spheroid implantation model. AB - OBJECTIVE: The orthotopic C6 glioma spheroid implantation model has been used to examine factors of neoangiogenesis, growth factor release, and protease expression as well the effect of antitumor agents. The present study systematically investigates the long-term course of orthotopically implanted C6 spheroid gliomas. METHODS: Reaggregated C6 spheroid tumors were implanted into the forebrain of 48 male Sprague-Dawley rats (32 immunocompetent, 16 thymectomized). The animals were examined by MRI at postoperative day (POD) 7, 14, 21, 28, 32, 45, 60, and 70. The MRI protocol included a T2-w and T1-w SE sequence before and after application of contrast medium and a CISS 3D sequence for volumetry. A total of six animals were selected after each MR exam from both groups and sacrificed for HE light microscopy and CD8+ T-lymphocyte, ED1+ macrophage, CD31+ endothelial cell immunohistochemistry. RESULTS: The tumors progressed to reach a maximum volume on day 28: 0.23 +/- 0.05 ml in the thymectomized and 0.16 +/- 0.021 ml in the immunocompetent group. Tumors then consistently regressed to vanish completely by POD 70. The influx of cytotoxic CD8+ T-lymphocytes correlated with tumor progression and the tumors reached a larger size in the thymectomized group. However, the time course of tumor regression was the same for both groups. CONCLUSION: The present data suggest that the orthotopic C6 glioma implanted into Sprague-Dawley rats will progress within a time span of approximately 4 weeks and can then retrogress again spontaneously. This finding has to be taken into account when deciding on a study protocol and the appropriate animal model. The C6 glioma model may be suitable to study the cell biological steps involved in the phenomenon of spontaneous tumor regression. PMID- 15530887 TI - Anti-cytokine autoantibodies in experimental autoimmune neuritis in Lewis rats. AB - Experimental autoimmune neuritis (EAN) is a CD4+ T-cell-mediated, inflammatory demyelinating disease of the peripheral nervous system (PNS) that serves as a model for Guillain-Barre syndrome (GBS) in humans. Cytokine production has been suggested to act a pathogenic role for EAN. To study the potential role of cytokines in context with cytokine autoantibodies (Aabs) in EAN, we used in situ hybridization to detect mRNA expression of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, IL-4, IL-10, and transforming growth factor (TGF)-beta in lymph node mononuclear cell (MNC) and in sciatic nerve sections, as well as ELISA for detection of their autoantibodies in sera and cerebrospinal fluid (CSF) over the course of EAN. Increased mRNA expression for IFN-gamma and TNF-alpha was registered correlating with the peak of clinical signs of EAN, and high levels of mRNA expression for IL-4, IL-10, and TGF-beta were associated with EAN recovery. The levels of cytokine mRNAs were generally inversely correlated to their autoantibodies in serum and CSF, whereby the CSF levels were equal to or lower than the serum levels. Autoantibodies to IFN-gamma dose-dependently inhibited IFN gamma-induced MHC expression by peritoneal macrophages proving a neutralizing biological effect of these autoantibodies. Our data demonstrate the existence of the anti-cytokine autoantibodies in the sera and CSF of rats with EAN; however, the role of anti-cytokine autoantibodies in the disease process of EAN remains to be resolved. PMID- 15530888 TI - The effect of brain-derived neurotrophic factor on sensory and autonomic function after lingual nerve repair. AB - Brain-derived neurotrophic factor (BDNF) is important in the response to peripheral nerve injury and may enhance regeneration. We have assessed its role in the functional recovery of sensory afferents and autonomic efferents after repair of the chorda tympani and lingual nerves in the cat. Six months after entubulation repair, with or without the incorporation of BDNF at the repair site, the recovery of secretomotor and vasomotor efferents was determined by recording salivary flow from the submandibular gland and temperature changes on the tongue surface, each evoked by stimulation of the repaired nerve. Electrophysiological recordings from the lingual and chorda tympani nerves proximal to the repair were undertaken to characterise mechanosensitive, thermosensitive, and gustatory afferents. When compared with data from uninjured control animals, both repair groups showed changes in receptor sensitivity and spontaneous discharge, and persistent reductions in conduction velocity, proportion of gustatory and thermosensitive units, rate of salivary secretion, and vasomotor responses. Comparisons between the outcome of repair with or without BDNF revealed few differences. In the BDNF group, fewer units in the chorda tympani responded to gustatory or thermal stimuli and the sensitivity of the gustatory units was lower. The conduction velocity of afferents in the lingual nerve was also lower, but the mechanoreceptive field size was higher. Thus, despite its known trophic role in the gustatory system, BDNF had not enhanced recovery of these or other fibre populations. We conclude that the application of BDNF to a site of lingual nerve repair has a negative effect on the long-term outcome. PMID- 15530889 TI - Role of dietary iron restriction in a mouse model of Parkinson's disease. AB - There is a growing body of evidence suggesting that iron chelation may be a useful therapy in the treatment of Parkinson's Disease (PD). Experiments were designed to test the impact of dietary iron availability on the pathogenic process and functional outcome in a mouse model of PD. Mice were fed diets containing low (4 ppm) or adequate (48 ppm) amounts of iron for 6 weeks before the administration of MPTP, a mitochondrial toxin that damages nigrostriatal dopaminergic neurons and induces Parkinson-like symptoms. Low dietary iron increased serum total iron binding capacity (P < 0.001). Consistent with neuronal protection, iron restriction increased sphingomyelin C16:0 and decreased ceramide C16:0. However, there was a 35% decrease in striatal dopamine (DA) in iron restricted mice. Motor behavior was also impaired in these animals. In vitro studies suggested that severe iron restriction could lead to p53-mediated neuronal apoptosis. Administration of MPTP reduced striatal DA (P < 0.01) and impaired motor behavior in iron-adequate mice. However, in iron-restricted mice, striatal dopamine levels and motor behavior were unchanged compared to saline treated mice. Thus, while reduced iron may provide protection against PD-inducing insults such as MPTP, the role of iron in the synthesis of DA and neuronal survival should be considered, particularly in the development of iron-chelating agents to be used chronically in the clinical setting. PMID- 15530890 TI - Alterations in expression of dopamine receptors and neuropeptides in the striatum of GTP cyclohydrolase-deficient mice. AB - The hph-1 mice have defective tetrahydrobiopterin biosynthesis and share many neurochemical similarities with l-dopa-responsive dystonia (DRD) in humans. In both, there are deficiencies in GTP cyclohydrolase I and low brain levels of dopamine (DA). Striatal tyrosine hydroxylase (TH) levels are decreased while the number of DA neurones in substantia nigra (SN) appears normal. The hph-1 mouse is therefore a useful model in which to investigate the biochemical mechanisms underlying dystonia in DRD. In the present study, the density of striatal DA terminals and DA receptors and the expression of D-1, D-2, and D-3 receptors, preproenkephalin (PPE-A), preprotachykinin (PPT), and nitric oxide synthase (NOS) mRNAs in the striatum and nucleus accumbens and nigral TH mRNA expression were examined. Striatal DA terminal density as judged by specific [3H]mazindol binding was not altered while the levels of TH mRNA were elevated in the SN of hph-1 mice compared to control (C57BL) mice. Total and subregional analysis of the striatum and nucleus accumbens showed that D-2 receptor ([3H]spiperone) binding density was increased while D-1 receptor ([3H]SCH 23390) and D-3 receptor ([3H]7-OH-DPAT) binding density was not altered. In the striatum and nucleus accumbens, expression of PPT mRNA was elevated but PPE-A mRNA, D-1, D-2 receptor, and nNOS mRNA were not changed in hph-1 mice compared to controls. These findings suggest that an imbalance between the direct strionigral and indirect striopallidal output pathways may be relevant to the genesis of DRD. However, the pattern of changes observed is not that expected as a result of striatal dopamine deficiency and suggests that other effects of GTP cyclohydrolase I deficiency may be involved. PMID- 15530891 TI - Caffeine-dependent stimulus-triggered oscillations in the CA3 region of hippocampal slices from rats chronically exposed to lead. AB - Yoshimura et al. [Yoshimura, H., Sugai, T., Onoda, N., Segami, N., Kato, N., 2002. Age-dependent occurrence of synchronized population oscillation suggestive of a developing functional coupling between NMDA and ryanodine receptors in the neocortex. Dev. Brain Res., 136, 63-68.] have shown that caffeine can elicit synchronized oscillations (10-12 Hz) dependent on calcium-induced calcium release in rat neocortex neurons. In the present work, synchronized oscillations in the CA3 region of rat hippocampus were studied by recording field excitatory postsynaptic potentials (fEPSPs) in vitro. In the presence of 0.1 mM caffeine, in CA3 of 44 of 45 (97.8%) slices from chronic lead-exposed rats, single electrical stimuli triggered a burst of high-frequency oscillations (approximately 230 Hz), whereas in CA3 of caffeine-treated slices from control rats, such oscillations could be elicited in only 2 of 24 (8.3%) slices. The complete (but fully reversible) block of caffeine-dependent oscillations by 6-cyano-7-nitro quinoxaline-2,3-dione (CNQX; 20 microM) indicates that alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionate (AMPA) receptors are necessary for the high frequency synchronized oscillations. 2-Amino-5-phosphonopentanoate (AP-5; 50 micoM) partially reduced the amplitude of caffeine-dependent oscillations without significantly altering their frequency. Caffeine-dependent oscillations could be abolished by application of AP-5 and 3 mM Mg2+ during the initial period of bursting, indicating that N-methyl-D-aspartate (NMDA) receptors play an important role in the generation of oscillations. The Ca2+ chelator ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA; 5 mM) added in standard artificial cerebrospinal fluid (ACSF) containing 0.1 mM caffeine fully blocked the oscillations. Caffeine-dependent oscillations are insensitive to an antagonist of gamma-aminobutyric acid (GABAA) receptors (10 microM bicuculline), L-type Ca2+ channels (10 muM nicardipine), L-type and N-type voltage-dependent calcium channels (100 microM Cd2)), and T-type Ca2+ channels (100 microM Ni2+). Previous studies have demonstrated that expression and function of NMDA and AMPA receptors are altered in the hippocampus of chronic lead-exposed rats. We propose that caffeine-dependent stimulus-induced oscillations in CA3 area of hippocampus from chronic lead-exposed rats are mainly mediated by the entry of extracellular Ca2+ through NMDA and non-NMDA receptors, without participation of GABAA receptors. Additionally, the underlying mechanisms are also discussed. PMID- 15530892 TI - Effects of isoflurane on prefrontal acetylcholine release and hypothalamic Fos response in young adult and aged rats. AB - This experiment investigated the influence of age on prefrontal acetylcholine (ACh) release and Fos response in the hypothalamic paraventricular nucleus and the nucleus tractus solitarius (NTS) of rats following isoflurane anesthesia. It is known that isoflurane decreases acetylcholine release in most brain regions. In the present study, we found that the level of prefrontal acetylcholine was significantly lower in 28-month-old rats (14% of baseline) than in 3-month-old rats (38% of baseline) during 2 h of isoflurane anesthesia (P < 0.05). The old rat group showed significantly greater Fos induction in the paraventricular nucleus compared to the young adult rat group (P < 0.05), indicating that the old rats were subjected to stress. No difference in Fos response was noted in the nucleus tractus solitarius. The old rats displayed a significant increase in feeding behavior during the 3-h recovery period (P < 0.05), but there was no difference in overall acetylcholine levels. Taken together, these findings suggest that isoflurane anesthesia influences old rats more profoundly than young adult rats with regard to reductions in acetylcholine release and stress responses. This may have implications for understanding the development of postoperative delirium in aged patients. PMID- 15530893 TI - Focally injected adenosine prevents seizures in the rat. AB - Prophylactic drug injection directly onto a seizure focus has the potential to improve seizure control with fewer side effects than is produced by systemic therapy. Using a dose-response model, we evaluated the effectiveness of adenosine application for focal seizure prophylaxis in 12 rats. Total spikes and electroencephalographic ictal events were reduced significantly by treatment with adenosine compared to control (P < 0.0001). This study demonstrates effectiveness and feasibility in a model system using intracranial injection of adenosine to prevent epileptiform events. PMID- 15530894 TI - Pharmacogenetics of HMG-CoA reductase inhibitors: exploring the potential for genotype-based individualization of coronary heart disease management. AB - Despite the benefit of statin therapy in the prevention of coronary heart disease, a considerable inter-individual variation exists in its response. It is well recognized that genetic variation can contribute to differences in drug disposition and, consequently, clinical efficacy at the population level. Pharmacogenetics, exploring genetic polymorphisms that influence response to drug therapy, may one day allow the clinician to customize treatment strategies for patients in order to improve the success rate of drug therapies. To date, 41 studies have investigated the relationships between common genetic variants and response to statin therapy in terms of lipid effects and clinical outcomes; 16 candidate genes involved in lipoprotein metabolism and 3 in pharmacokinetics. APOE is the most extensively studied locus, and absolute difference in LDL cholesterol reduction across genotypes remained 3-6%. Moreover, none of the associations was striking enough to justify genetic analysis in clinical practice. Reported data have suggested that larger studies (>1000 participants) or combination analyses with >2 different polymorphisms would enable us to find clinically or biologically meaningful difference, which could be assumed as >10% absolute difference, and that genes influencing cholesterol biosynthesis in the liver, such as ABCG5/G8, CYP7A1, HMGCR, would be good candidates for future studies. PMID- 15530896 TI - Chlamydia pneumoniae (C. pneumoniae) infection upregulates atherosclerosis related gene expression in human umbilical vein endothelial cells (HUVECs). AB - There is accumulating evidence that supports a role of infection in atherosclerosis, with possible mechanism by injuring to the endothelium and inducing an autoimmune response to heat shock proteins (HSPs). In this study, a cDNA array, containing 588 human cardiovascular genes, was utilized to analyze the gene expression profile of Chlamydia pneumoniae (C. pneumoniae) infected human umbilical vein endothelial cells (HUVECs). After 48h of C. pneumoniae infection, the HUVECs were harvested and subjected to immunofluorescent staining, electron microscopy, cDNA array hybridization, RT-PCR, and immunoblotting. This study found a panel of human host genes that were upregulated by C. pneumoniae. The majority of these genes were related to complex lipid metabolism, adhesion receptors, hormones, hormone receptors, and a metalloproteinase that may contribute to atherosclerosis in vivo. Representatives of upregulated gene products, i.e., heat shock protein 60 (HSP60), macrophage scavenger receptor, cytochrome P450, and VEGF165R were immunofluorescently detected in HUVECs, with their greater expression induced by C. pneumoniae infection. These findings supported the opinion that C. pneumoniae might contribute to atherosclerotic development in vivo. PMID- 15530895 TI - Sulfone COX-2 inhibitors increase susceptibility of human LDL and plasma to oxidative modification: comparison to sulfonamide COX-2 inhibitors and NSAIDs. AB - Clinical investigations have demonstrated a link between use of the sulfone cyclooxygenase-2 (COX-2) inhibitor, rofecoxib, and increased risk for atherothrombotic events. This increased risk was not observed for a sulfonamide COX-2 inhibitor (celecoxib), indicating a potential non-enzymatic mechanism for rofexocib. To test this hypothesis, we compared the independent effects of COX-2 inhibitors on human LDL oxidation, an important contributor to atherosclerotic cardiovascular disease. The results showed that rofecoxib (100 nM) significantly decreased (>40%, p<0.001) the lag time for LDL conjugated diene formation and increased levels of thiobarbituric-acid-reactive-substances (TBARS) in vitro. The pro-oxidant activity of rofecoxib was dose-dependent and attenuated by 70% (p<0.001) with the antioxidant, Trolox. Rofecoxib and etoricoxib (100 nM) also caused a marked increase (>35%, p<0.001) in non-enzymatic generation of isoprostanes, as measured by mass spectroscopy. Addition of rofecoxib to fresh human plasma reduced the oxygen radical antioxidant capacity (ORAC) by 34% (p<0.0001). By contrast, other selective (celecoxib, valdecoxib, meloxicam) and non-selective COX inhibitors (ibuprofen, naproxen, diclofenac) had no significant effect on LDL oxidation rates or plasma ORAC values, even at suprapharmacologic levels. X-ray diffraction analysis showed that sulfone COX-2 inhibitors interact differently with membrane phospholipids, suggesting a physico-chemical basis for the pro-oxidant activity. These results demonstrate that sulfone COX-2 inhibitors increase the susceptibility of biological lipids to oxidative modification through a non-enzymatic process. These findings may provide mechanistic insight into reported differences in cardiovascular risk for COX-2 inhibitors. PMID- 15530897 TI - Fenofibrate enhances CD36 mediated endocytic uptake and degradation of oxidized low density lipoprotein in adipocytes from hypercholesterolemia rabbit. AB - BACKGROUND: CD36 as a fatty acid transporter is predominantly expressed in adipocytes. We studied whether adipocytes could uptake and degrade OxLDL through CD36 and explored the effect of fenofibrate on OxLDL uptake in adipocytes from hypercholesterolemia rabbits. METHODS: Subcutaneous adipose tissues were collected from normal, high-cholesterol and high-cholesterol plus fenofibrate treatment rabbits for adipocytes culture. CD36 and peroxisome proliferator activated receptor gamma (PPARgamma) mRNA expression were evaluated by RT-PCR. RESULTS: Cellular expression of CD36 was confirmed during differentiation of adipose cell by RT-PCR. Upon incubation at 37 degrees C, (125)I-OxLDL was endocytosed in a dose-dependent fashion and underwent lysosomal degradation by adipocytes. In binding experiments at 4 degrees C, (125)I-OxLDL exhibited specific and saturable binding to adipocytes (K(D) = 4.2 microg/mL). The endocytic uptake and degradation of (125)I-OxLDL by adipocytes were inhibited by 56 and 54% with anti-CD36 antibody. Fenofibrate treatment enhanced the (125)I OxLDL uptake and degradation and up-regulated CD36 mRNA expression in adipocytes and suppressed PPARgamma mRNA expression in adipose tissue from hypercholesterolemia rabbits. CONCLUSION: CD36 plays a novel role in adipose tissues and adipocytes possibly involve in clearance of OxLDL in blood. Fenofibrate treatment improved the OxLDL uptake and degradation in adipocytes from hypercholesterolemia rabbits. PMID- 15530898 TI - Upregulation of lectinlike oxidized low-density lipoprotein receptor-1 expression contributes to the vein graft atherosclerosis: modulation by losartan. AB - Vein grafts interposed to arteries are susceptible to the development of accelerated atherosclerosis. The role of lectinlike oxidized LDL (ox-LDL) receptor-1 (LOX-1) expression in the atherosclerotic lesions of vein grafts has not been clarified. The current study was designed to examine the expression of LOX-1 in vein grafts atherosclerosis and the modulating effect of losartan on it. Autologous external jugular veins were grafted to common carotid arteries in 30 male New Zealand White rabbits. After surgery, rabbits were fed with high cholesterol diet (HC), high cholesterol diet plus losartan (10 mg/kg/day, LHC) or regular chow (control, n = 10 in each group) for 12 weeks. LOX-1 expressions in the grafts were examined by immunohistochemistry, semi-quantitative RT-PCR and Western blot. Neointimal hyperplasia was observed in all vein grafts characterized by extensive intimal thickening. Atherosclerotic lesions were found in vein grafts of HC group, which were attenuated by losartan. Losartan also reduced the vein grafts atherosclerotic plaque vulnerability. LOX-1 expression was low in the endothelium and neointima of vein grafts in control group and was significantly increased in the endothelium and atherosclerotic lesions in HC group but not in LHC group. In conclusion, LOX-1 was expressed in endothelium and neointima of autologous vein grafts of rabbits. Increased LOX-1 expression was associated with vein grafts atherosclerosis development. Downregulation of LOX-1 by losartan might contribute to its attenuating effect on vein grafts atherosclerosis. PMID- 15530899 TI - Low flow promotes instent intimal hyperplasia. Comparison with lumen loss in balloon-injured and uninjured vessels and the effects of the antioxidant pyrrolidine dithiocarbamate. AB - Low flow (LF) promotes late lumen loss after angioplasty by exacerbating inward remodelling through redox-sensitive mechanisms. Stents eliminate inward remodelling and the effect of LF on in-stent restenosis is uncertain. We performed over-sized (1.3-1.5:1) stenting (S) and balloon injury (in the same vessel, B) to the carotid arteries of cholesterol-fed rabbits and compared 28-day late lumen loss with that in an uninjured segment in the same vessel (U). Vessels (n = 5 animals per group) were subjected to high (H), normal (N) and low (L) flow in animals fed either vehicle (V) or the antioxidant pyrrolidine dithiocarbamate, PDTC (P). LF significantly increased in-stent neointima formation relative to normal and high flow (SLV 0.72 +/- 0.07 mm(2) versus SNV 0.43 +/- 0.08 mm(2) versus SHV 0.28 +/- 0.04 mm(2), P < 0.05). However, LF resulted in greater lumen loss in segments from the same vessel subject to balloon injury (lumen SLV 5.18 +/- 0.40 mm(2) and SNV 5.32 +/- 0.40 mm(2) versus BLV 1.28 +/- 0.33 mm(2) and BNV 2.19 +/- 0.28 mm(2)), by greater enhancement of inward remodelling. In addition, inward remodelling and lumen loss due to LF were greater in balloon-injured segments than in adjacent uninjured segments where shear homeostatic remodelling occurs (lumen BLV 1.28 +/- 0.33 mm(2) versus ULV 1.52 +/- 0.22 mm(2)). Lastly, while PDTC effectively reduced intima formation and inward remodelling due to LF in balloon-injured vessels there was no effect on flow-dependent neointima formation in stented vessels. We conclude that LF accentuates in-stent neointima formation, but that flow-dependent lumen loss after stenting is less than that after balloon injury. When LF is present lumen loss can be minimised by antioxidants or stenting. PMID- 15530900 TI - Linoleic acid increases monocyte deformation and adhesion to endothelium. AB - Fatty acids have been implicated in having both anti- or pro-inflammatory actions, which may contribute to the progression and severity of atherosclerosis. Linoleic acid has been shown by others to decrease CD18 expression and leukocyte adhesion under static conditions. We investigated the effect of steric acid (18:0), oleic acid (18:1), and linoleic acid (18:2) on the cortical tension (a measure of cell membrane deformability) and adhesion characteristics of the monocytic cell line Mono Mac 6 (MM6) cells to TNF-alpha activated HUVEC under fluid flow. Linoleic acid concentrations up to 23 microM decreased cortical tension and increased adhesion frequencies. Increased adhesion was not due to altered cell morphology or adhesion kinetics and occurred despite decreases in receptor expression (CD18 and CD11a). At higher levels of linoleic acid (> or = 46 microM), cell dissociation constants significantly increased. Results show that decreasing cortical tension increased the probability that contact between MM6 cells and endothelium would produce an adhesive interaction, possibly due to increased deformation of the microvilli and the cell membrane cortex. However, more deformable cells rolled more erratically at low shear rates. The different behavior during initial contact and rolling suggest that adhesion is influenced by two force-dependent mechanisms, deformation of microvilli and a steric barrier. Incubation of MM6 with 23 microM steric or oleic acid did not significantly affect cortical tension. However, cells incubated with steric acid greatly increased their adherence to HUVEC and cells incubated with oleic acid showed no significant effect, indicating factors other than deformability may dominate. PMID- 15530901 TI - Identification of genes predominantly expressed in human macrophages. AB - Identification of cell and tissue specific genes may provide novel insights to signaling systems and functions. Macrophages play a key role in many diseases including atherosclerosis. Using DNA microarrays we compared the expression of approximately 10,000 genes in 56 human tissues and identified 23 genes with predominant expression in macrophages. The identified genes include both genes known to be macrophage specific and genes previously not well described in this cell type. Tissue distribution of two genes, liver X receptor (LXR) alpha and interleukin-1 receptor antagonist (IL1RN), was verified by real-time RT-PCR. We conclude that comparison of expression profiles from a large number of tissues can be used to identify genes that are predominantly expressed in certain tissues. Identification of novel macrophage specific genes may increase our understanding of the role of this cell in different diseases. PMID- 15530902 TI - Absence of an atheroprotective effect of the garlic powder printanor in APOE*3 Leiden transgenic mice. AB - Numerous animal studies have reported that garlic can protect against atherosclerosis. However, a comparable number of studies do not support this observation. This contradiction may result from differences in study design, use of different animal models, and use of different garlic formulations and preparations. Here, we investigated the effect of the chemically well characterized and production-controlled garlic powder printanor on atherosclerosis in the APOE*3-Leiden transgenic mouse, a mouse model well suited for evaluating anti-atherosclerotic properties of drugs and food components under human-like conditions. APOE*3-Leiden mice were fed a Western diet supplemented with either 5 or 50 g kg(-1) printanor. As a reference, the commercially available fermented garlic kyolic was included (1.6 g kg(-1) diet). Treatment with printanor demonstrated reduced body weight, coinciding with increased feces production and fecal fatty acids excretion. Printanor and kyolic treatment did not affect plasma lipids, markers of inflammation (serum amyloid A, serum-soluble intercellular adhesion molecule-1, and blood-leukocytes tumor necrosis factor alpha (TNFalpha) production) and vascular activation (plasma von Willebrand factor (vWF)). As analyzed after 28 weeks of treatment, printanor and kyolic did not affect atherosclerotic lesion type, area or composition. Under conditions relevant to the human situation, the well-characterized and production-controlled garlic powder printanor does not display hypolipidemic, anti-inflammatory or anti atherosclerotic properties. PMID- 15530903 TI - Increased lysophosphatidylcholine and non-esterified fatty acid content in LDL induces chemokine release in endothelial cells. Relationship with electronegative LDL. AB - Electronegative low-density lipoprotein (LDL(-)) is a plasma-circulating LDL subfraction with proinflammatory properties that induces the production of chemokines in cultured endothelial cells. However, the specific mechanism of LDL( )-mediated chemokine release is presently unknown. A characteristic feature of LDL(-) is an increased content of lysophosphatidylcholine (LPC) and non esterified fatty acids (NEFA). The effect of increasing amounts of LPC and NEFA associated with LDL on the release of chemokines by endothelial cells was studied. Total LDL was subfractionated by anion-exchange chromatography in electropositive (LDL(+)) and LDL(-). LDL(-) contained two-fold more LPC and NEFA than LDL(+) and induced two- to four-fold more (p < 0.05) interleukin-8 (IL-8, 11.5 +/- 8.2 ng/10(5) cells) and monocyte chemotactic protein-1 (MCP-1, 10.8 +/- 3.8 ng/10(5) cells) release by human umbilical vein endothelial cells (HUVEC) than LDL(+) (IL-8: 3.4 +/- 1.5 ng/10(5) cells, MCP-1: 5.8 +/- 2.9 ng/10(5) cells). The content of LPC and NEFA in LDL(+) was increased by enzymatic treatment with secretory phospholipase A(2) (sPLA(2)) at 5 ng/mL or 20 ng/mL or by incubation with NEFA at 2 mmol/L. Modification of LDL(+) by both methods did not result in oxidative modification as demonstrated by the lack of change in antioxidants, conjugated dienes and malondialdehyde content. sPLA(2) treatment resulted in an increase in LPC and NEFA in LDL(+) which enhanced its ability to release IL-8 and MCP-1 by HUVEC in a concentration-dependent manner (sPLA(2)(5) LDL; IL-8: 7.1 +/- 3.8ng/10(5) cells, MCP-1: 8.0 +/- 5.1 ng/10(5) cells; sPLA(2)(20)-LDL; IL-8: 20.8 +/- 11.2 ng/10(5) cells, MCP-1: 15.0 +/- 7.5 ng/10(5) cells). NEFA loading of LDL(+) also favored the release of IL-8 and MCP-1 (IL-8: 7.8 +/- 6.1 ng/10(5) cells, MCP-1: 8.4 +/- 2.7 ng/10(5) cells, p < 0.05 versus LDL(+)). These effects were observed when modified LDL(+) reached a content of LPC and/or NEFA similar that of LDL(-). These data indicate that non-oxidized polar lipids associated with LDL promote an inflammatory response in endothelial cells and suggest that increased NEFA and LPC could be involved in the inflammatory activity of LDL(-). PMID- 15530904 TI - Fenofibrate lowers adiposity and corrects metabolic abnormalities, but only partially restores endothelial function in dietary obese rats. AB - In humans, dietary-induced obesity markedly increases plasma lipid profile and impairs vascular function leading to increased incidence of cardiovascular events. We have recently reported that chronic withdrawal of obesity-inducing diet attenuates obesity and completely corrects endothelial function. The aim of this study was to investigate whether fenofibrate-induced decrease in adiposity would also correct vascular function in the presence of obesity-inducing diet. Wistar rats were fed with either standard laboratory chow (lean, n = 9) or given a highly palatable diet (diet-fed, n = 18) for 15 weeks. After 7 weeks, half of the diet-fed group was treated with fenofibrate (fenofibrate-treated, n = 9) for 8 weeks before being sacrificed. Untreated diet-fed (n = 9) rats had significantly higher body weight, total fat mass (by up to two-fold, p < 0.001 for both), and raised fasting plasma levels of insulin, leptin and triglycerides (up to 110%; p < 0.001), but not glucose or nonesterified fatty acids (NEFA) than both lean control and fenofibrate-treated groups. Resistance mesenteric arteries responses to KCl- and noradrenaline-induced vasoconstriction were similar in all three groups. However, compared with lean controls, endothelium-dependent vasorelaxation responses were shifted to the right in both untreated and fenofibrate-treated diet-fed groups. Fenofibrate treatment improved endothelium dependent vasorelaxation at only high carbamycholine concentrations (10 microM). There were no differences in endothelium-independent vasorelaxation between the three groups. These results indicate that, in the presence of obesity-inducing diet, fenofibrate markedly reverses obesity and corrects insulin resistance and lipid profile, but it only has a limited beneficial effect on vascular function. Therefore, it seems that diet component rather than obesity per se plays a key role in the genesis of vascular abnormalities. PMID- 15530905 TI - Glycoxidized low-density lipoprotein regulates the expression of scavenger receptors in THP-1 macrophages. AB - Low-density lipoprotein (LDL) in patients with diabetes is subject to modification by both oxidation and glycation. In contrast to oxidized LDL, the biological effects of glycoxidized LDL have not been well characterised. In this study, the effects of oxidized, glycated, glycoxidized and oxidized LDL on scavenger receptor gene expressions, and the induction of oxidized LDL uptake and cholesteryl ester accumulation in THP-1 macrophages were compared. Modified LDL was incubated with THP-1 macrophages. Gene expression of scavenger receptor class A (SR-A), CD36 and scavenger receptor class B type I (SR-BI) was determined by quantitative reverse transcriptase PCR (RT-PCR). Glycoxidized LDL was able to significantly induce SR-A and CD36 expression by 3- and 4.5-fold, respectively, in macrophages whereas SR-BI expression was suppressed by glycoxidized LDL, glycated LDL and oxidized LDL. Incubation with glycoxidized LDL enhanced the uptake of DiI-labeled oxidized LDL by macrophages to a greater extent than that of glycated LDL or oxidized LDL. Glycoxidized LDL also induced a significant degree of intracellular cholesteryl ester accumulation. Taken together, our results would suggest that glycoxidized LDL might be an important candidate in the initiation of foam cell formation and might play a significant role in the pathogenesis of atherosclerosis in diabetes mellitus. PMID- 15530907 TI - Small low-density lipoprotein particles and endothelium-dependent vasodilation in postmenopausal women. AB - Low concentrations of estrogen may decrease endothelial function in postmenopausal women. Elevated plasma triglycerides after menopause are frequently associated with a small, dense low-density lipoprotein (LDL) phenotype. Small LDL particles that are more susceptible to oxidation can also inhibit endothelium-dependent vasodilation. The purpose of the present study was to investigate whether hypertriglyceridemia-induced small LDL particles are associated with endothelial dysfunction in postmenopausal women. We studied 15 premenopausal and 41 postmenopausal women. Postmenopausal subjects were divided into those with LDL subclass pattern A (large particles) and those with pattern B (small particles). Plasma lipids, hormones, and diameter and oxidative susceptibility of LDL were measured. Vasodilatory responses of the brachial artery were evaluated by measuring flow-mediated vasodilation (FMD) and nitroglycerin-induced vasodilation (NID). FMD in both postmenopausal groups was significantly lower than in premenopausal women. FMD in subjects with pattern B was significantly smaller than in those with pattern A (4.9 +/- 1.9% versus 8.8 +/- 3.6%). NID did not differ significantly among the groups. Plasma triglyceride concentrations were higher, lag time for LDL oxidation was shortened, and LDL derived thiobarbituric acid-reactive substance (TBARS) concentrations were significantly greater in subjects with pattern B than in premenopausal or pattern A subjects. LDL diameter correlated negatively with plasma triglycerides (r = 0.51) or LDL-derived TBARS (r = -0.44) and positively with LDL-lag time (r = 0.66). FMD correlated negatively with LDL-derived TBARS (r = -0.36) and positively with LDL diameter (r = 0.44) or LDL-lag time (r = 0.43). Vascular endothelial dysfunction may be associated with elevated triglyceride-induced small LDL particles that have enhanced oxidative susceptibility in postmenopausal women. PMID- 15530906 TI - Heat shock protein 60 autoimmunity and early lipid lesions in cholesterol-fed C57BL/6JBom mice during Chlamydia pneumoniae infection. AB - Chronic Chlamydia pneumoniae infection and autoimmunity to heat shock protein 60 (Hsp60) have both been documented to be associated with atherosclerosis. Herein, we studied the effects of C. pneumoniae infection and a diet with a low cholesterol supplement on the development of autoantibodies to mouse Hsp60 and early lipid lesions in the aortic valve of C57BL/6JBom mice. In addition, pulmonary infection was investigated. C57BL/6JBom mice were given one to three C. pneumoniae inoculations and fed either a regular diet or a diet enriched with 0.2% cholesterol. Autoantibody responses against mouse Hsp60 developed in both diet groups when the mice were infected with C. pneumoniae and in uninfected mice fed a cholesterol-enriched diet. C. pneumoniae infections increased subendothelial foam cell accumulation in mice on a 0.2% cholesterol-enriched diet (p = 0.022), without apparent hypercholesterolemia. These in vivo data suggest that autoantibodies against mouse Hsp60 develop as a consequence of cholesterol feeding and repeated C. pneumoniae infections. Further, infectious burden increased early lipid lesions in C57BL/6JBom mice fed a cholesterol-enriched diet. PMID- 15530908 TI - Total homocysteine is not a determinant of arterial pulse wave velocity in young healthy adults. AB - AIMS: Hyperhomocysteinaemia has been associated with reduced pulse wave velocity (PWV) in patients with end-stage renal disease and in those with hypertension. The aim of this study was to examine the association between total homocysteine (tHcy) concentrations, the biochemical and genetic determinants of tHcy and PWV in healthy young adults. METHODS AND RESULTS: A total of 489 subjects aged 20-25 years participated. A fasting blood sample was taken and PWV measured using a non invasive optical method. tHcy did not correlate with PWV, whether assessed at the aorto-iliac segment (P = 0.18), the aorto-radial segment (P = 0.39) or the aorto dorsalis-pedis segment (P = 0.22). When tHcy was classified into normal (<15) and high (> or =15micromol/l), PWV did not differ between the two groups at any segment. PWV did not differ by MTHFR C677T or NOS3 G894T genotype, even when smoking and folate sub-groups were considered. Considering aortic PWV as a dependent variable, stepwise regression analysis showed that the only parameter entering the model for all segments was systolic blood pressure (aorto-iliac, P < 0.001; aorto-radial, P = 0.01; aorto-dorsalis-pedis, P = 0.001). Age, sex, COL1A1 genotype and triglycerides entered the model significantly for two of three segments. CONCLUSION: This study shows that arterial PWV is not associated with tHcy in a healthy young population. PMID- 15530909 TI - New and confirmatory evidence of an association between APOE genotype and baseline C-reactive protein in dyslipidemic individuals. AB - We have investigated the association between APOE genotypes and C-reactive protein (CRP) levels in a cohort of approximately 600 individuals who were candidates for statin therapy. An association had been previously reported between the APOE3 allele and elevated CRP levels. That study only examined men. We have reproduced that association in men and have extended the finding to women. We also investigated the effect of the interaction between APOE genotype and hormone replacement therapy (HRT) status on CRP levels, adjusting for body mass index (BMI) and other covariates. BMI and HRT are also significant predictors of CRP, as previously reported. The effect of HRT is strong enough that the contribution of APOE genotype is no longer statistically significant among women on HRT. We also demonstrate that the presence or absence of the single SNP Cysl30Arg (which distinguishes APOE4 from APOE2 and APOE3) is sufficient to determine whether an individual is predisposed to higher or lower CRP levels. Essentially, the presence of one or two copies of APOE4 is associated with a reduction of CRP levels by approximately 34% relative to individuals with zero copies (1.73 mg/L for subjects with one or two copies versus 2.63 mg/L for subjects with zero copies of APOE4). We also tested previously reported associations between CRP levels and polymorphisms in the CRP and IL6 genes. These associations were not reproduced in our cohort. PMID- 15530910 TI - Application of real-time RT-PCR to quantifying gene expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in human abdominal aortic aneurysm. AB - BACKGROUND: The relative expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs), key regulators in remodeling of extracellular matrix, are considered to play a pivotal role in the development of abdominal aortic aneurysm (AAA). However, few data exist regarding quantitative assessment of their expression in clinical settings. METHODS: In 22 patients with AAA who underwent graft replacement, tissue samples of the AAA and non-dilated aorta were obtained. Using a real-time RT-PCR method that enabled quantitative measurement of mRNA levels in small tissue samples, we determined gene expression levels of MMPs and TIMPs relative to that of glutaraldehyde 3 phosphate dehydrogenase in each sample. RESULTS: The expression levels of the MMP 1 and -3 genes were significantly augmented in AAA compared with non-dilated regions (4.48 +/- 2.01 versus 0.26 +/- 0.12, P < 0.01 and 1.89 +/- 1.00 versus 5.01 +/- 0.97, P < 0.05, respectively). Although genes for TIMP-1, -2 and -3 tended to be upregulated in AAA, relative expression levels of MMP-1 to TIMP-1, MMP-1 to TIMP-2, MMP-1 to TIMP-3, and MMP-3 to TIMP-2 were still higher in AAA than in non-dilated regions (1.12 +/- 0.63 versus 0.10 +/- 0.03, 4.13 +/- 1.12 versus 0.43 +/- 0.11, 1.61 +/- 0.59 versus 0.14 +/- 0.03, and 7.81 +/- 1.60 versus 2.56 +/- 0.76, respectively, P < 0.05). CONCLUSION: These results demonstrate that the present real-time RT-PCR method is reliable for the determination of mRNA levels in small samples of vascular tissue and that disproportional expression of both MMP-1 and MMP-3 relative to TIMPs relates pathologically to the evolution of AAA. PMID- 15530911 TI - Lecithin: cholesterol acyltransferase (LCAT) deficiency and risk of vascular disease: 25 year follow-up. AB - We have reassessed the clinical and biochemical status of a large Canadian kindred with LCAT deficiency 25 years after the initial investigations. There have been no vascular events or death in this family over the 25 years. Both the homozygous (N = 2) and heterozygous (N = 9) patients had highly abnormal lipid profiles with low HDL-C (extreme in the homozygotes); apo B levels were high in the heterozygotes. Lipoprotein and hepatic lipase activities were low in the homozygotes and several heterozygotes. In the two homozygotes the carotid intima media thickness (IMT) was above 75th percentile expected for age and gender. However, the IMT abnormalities were much more pronounced in the heterozygotes, four of whom also had detectable plaques. The homozygotes had only minimal increases in IMT, no plaques, no IMT changes over the last 4 years and normal endothelial function. We conclude that, in this kindred, no significant vascular changes were observed in the homozygotes. However, heterozygocity for LCAT deficiency is associated with both an atherogenic lipid profile and vascular abnormalities. PMID- 15530912 TI - Apolipoprotein B determines risk for recurrent coronary events in postinfarction patients with metabolic syndrome. AB - The adult treatment panel III (ATP III) of the National Cholesterol Education Program recognizes metabolic syndrome (MS) as a secondary target for risk reduction therapy given the well-known increase in risk of cardiovascular disease (CVD) by all MS components. We investigated potential CVD risk arising from 17 laboratory variables in postinfarction patients with MS by identifying such patients from the Thrombogenic Factors and Recurrent Coronary Events (THROMBO) study using slightly modified ATP III criteria for MS. This gave 272 patients with MS out of a total of 766 postinfarction patients without history of diabetes. Comparison between non-MS and MS patients using Kaplan-Meier analysis revealed no difference in outcome between groups. Additionally, a proportional hazards model applied to dichotomized laboratory parameters demonstrated only apoB as a significant predictor of risk for recurrent coronary events in MS patients with hazard ratio, 1.97 (95% CI; 1.08, 3.60; P < 0.05). We conclude that there is no difference in outcomes between non-MS and MS postinfarction patients; that apoB is significantly associated with risk for recurrent coronary events in postinfarction patients with MS; that further studies would be needed to recommend the routine determination of apoB in these patient groups. PMID- 15530913 TI - Homocysteine as a risk factor for coronary heart diseases and its association with inflammatory biomarkers, lipids and dietary factors. AB - The causal relation of total Homocysteine (tHcy) to coronary heart diseases (CHD) is unclear. In vitro studies suggest a proinflammatory effect. Among 32,826 women from the Nurses' Health Study who provided blood samples in 1989-1990, 237 CHD events were documented during 8 years of follow-up. The cases (1:2) were matched to controls on age, smoking, and month of blood draw. Plasma tHcy was inversely associated with blood levels of folate (partial r = -0.3, P < 0.0001) and B1(2) (r = -0.2, P < 0.0001) and with dietary intake of folate (r = -0.1, P < 0.01) and B(2) vitamin (r = -0.1, P = 0.01). tHcy was positively associated with soluble tumor necrosis receptor (sTNF-R) 1 and 2 (partial r = 0.2, P < 0.0001). In a multivariate model adjusted for age, smoking, BMI, parental history, hypertension, diabetes, postmenopausal hormone use, physical activity and alcohol intake, the relative risk of CHD between the extreme quartiles of tHcy was 1.66 (95% CI; 1.05-2.64, P trend = 0.02). The association was not appreciably attenuated after further adjustments for sTNF-R1, sTNF-R2, CRP, or Total Cholesterol:/HDL-c ratio. tHcy is an independent risk predictor of CHD and modestly associated with TNF-receptors. However, the inflammatory biomarkers measured could not explain its role in CHD. PMID- 15530914 TI - Association of high serum concentration of the third component of complement (C3) with pre-existing severe coronary artery disease and new vascular events in women. AB - Atherosclerosis is an inflammatory disease. The complement system plays an important role in the atherosclerotic process. However, lesser data is available on the possible role of C3 as a risk factor for atherosclerosis. Therefore, in a follow up study we determined C3 levels in 266 patients with pre-existing severe coronary artery disease (CAD) and compared their serum C3 concentrations with the cause of the disease. We investigated whether C3 levels predict the major complications of severe CAD during a 5-year long follow up period in patients, who have received an aorto-coronary bypass graft surgery. C3 concentrations were elevated in the patients with severe CAD compared to 182 healthy controls, and women had higher C3 concentrations than men. Pathological C3 levels (C3> or =1.8 g/L) were able to predict major complications of atherosclerosis (death by cardiac events, new acute myocardial infarction, stroke, carotid surgery and peripheral arterial disease) that developed during the follow up period only in women (OR: 4.1, 95% C.I. 1.23-13.61, p = 0.0249) independent of other risk factors for atherosclerosis. Our data supports the assumption that high C3 indicates the progression of atherosclerosis as a special marker of chronic inflammation. PMID- 15530915 TI - LDL size, total antioxidant status and oxidised LDL in normal human pregnancy: a longitudinal study. AB - The aim of our work was to evaluate changes in levels of oxidised low-density lipoprotein (Ox-LDL) during pregnancy and how they correlate with changes in LDL size and serum total antioxidant status (TAS). LDL peak and mean particle diameter (LDL-PPD and LDL-MPD, respectively) and the relative proportion of 3 LDL subfractions were quantified. We evaluated plasma levels of Ox-LDL and serum levels of TAS, total cholesterol (Chol), triglycerides (TG), apolipoprotein A-I (apo A-I), apolipoprotein B (apo B), HDL-cholesterol (HDLc) and LDL-cholesterol (LDLc). A longitudinal study was performed in the three trimesters (T1-T3) of pregnancy in normal pregnant women (n = 23) and a non-pregnant group (n = 18) was used as control. TG levels were significantly elevated whereas LDL-MPD and LDL PPD were significantly reduced in T1 compared to controls. Ox-LDL, TG, Chol, apo B and LDLc rose markedly throughout pregnancy with significant changes between each trimester; LDL-PPD, LDL-MPD and TAS levels decreased significantly from T1 to T3. Changes in LDL size and in Ox-LDL and TAS levels were more pronounced between T1 and T2 than between T2 and T3. HDLc and apo A-I reached peak concentration in T2 but decreased in T3. TG concentrations correlated inversely with LDL size and positively with Ox-LDL; Ox-LDL was positively and strongly correlated with LDLc. Moreover, relative changes in the levels of Ox-LDL correlated inversely with relative changes in LDL size and TAS between trimesters. In conclusion, during human gestation the change in LDL profile towards smaller species and the decrease in serum TAS are closely associated with increased levels of Ox-LDL. The exact physiological role of the increments in Ox LDL during pregnancy remains to be clarified. PMID- 15530916 TI - The effect of acute red wine polyphenol consumption on postprandial lipaemia in postmenopausal women. AB - Postprandial lipoproteins are potentially atherogenic. The aim of this study was to elucidate whether acute consumption of red wine (RW) and dealcoholised red wine (DRW) regulates postprandial lipid and lipoprotein metabolism in 17 dyslipidaemic postmenopausal women. A mixed meal accompanied by either water, RW or DRW was consumed on three separate visits, in random order, 2 weeks apart. One fasting and 6 hourly postprandial blood samples were taken for lipid analysis. Results showed no significant quantitative changes in postprandial apolipoprotein (apo) B48 levels following the consumption of DRW or RW compared to water. However, qualitatively, DRW may reduce arterial exposure to apoB48-containing lipoproteins over the 6-h postprandial period measured. DRW consumption did not significantly change postprandial TG or insulin levels. A 35% (p = 0.02) increase in postprandial triglyceride (TG) levels and a 54% (p = 0.02) increase in insulin levels were observed following RW consumption, compared to water. In conclusion, acute DRW consumption had no effect on postprandial lipid and lipoprotein metabolism in dyslipidaemic postmenopausal women. However, the consumption of full-compliment RW exacerbated the postprandial lipaemic and insulin response over the 6-h period. Collectively, our findings suggest that neither polyphenols nor red wine reduce atherosclerotic risk by acutely modulating postprandial lipaemia over a 6-h period. PMID- 15530917 TI - The A-1087IL-10 allele is associated with cardiovascular disease in SLE. AB - The risk of cardiovascular disease (CVD) in SLE patients is very high. It is therefore surprising that IL-10 has been discussed both as pathogenic in SLE and as an atheroprotective cytokine. In contrast, TNF is believed to be atherogenic and we recently reported that raised activity in the TNF-system is implicated in SLE-related CVD. Twenty-six (aged 52 +/- 8 years) female patients with SLE and a history of CVD (myocardial infarction, angina, stroke or claudication) were compared with 26 age-matched SLE patients without CVD (SLE controls) or 26 age matched population controls. The -1087IL-10 gene polymorphism was determined by PCR with restriction endonuclease mapping. Serum IL-10 and TNF-levels were determined by ELISA. The A allele frequency of -1087IL-10 gene in SLE/CVD was higher than in SLE controls (0.62 versus 0.42, p < 0.05). Ten (38%) of 26 SLE/CVD exhibited IL-10 AA genotype compared with five (19%) of 26 SLE controls. Serum IL 10 and TNF-levels were raised in SLE/CVD compared with SLE controls or population controls (p < 0.001). Furthermore, in SLE/CVD, a significantly reduced IL-10:TNF ratio was observed in patients with IL-10 AA genotype compared with AG or GG genotype (0.56 versus 0.77 versus 1.24, p < 0.05). In SLE controls and population controls, individuals with IL-10 GG genotype tended to have higher IL-10:TNF ratio. In conclusion, the A-1087IL-10 allele which has been reported to cause a lower capacity for IL-10 production could contribute to CVD in SLE. Furthermore, the IL-10 AA genotype is associated with reduced ratio of atheroprotective to atherogenic cytokines in SLE patients with CVD. PMID- 15530918 TI - No genetic linkage or molecular evidence for involvement of the PCSK9, ARH or CYP7A1 genes in the Familial Hypercholesterolemia phenotype in a sample of Danish families without pathogenic mutations in the LDL receptor and apoB genes. AB - A locus on chromosome 1p34.1-p32 has been linked to autosomal dominant Familial Hypercholesterolemia (FH) and is termed the third FH locus. We tested whether this third FH locus is linked to the FH phenotype in 20 Danish families, with 158 members, without pathogenic mutations in the genes, encoding the low-density lipoprotein (LDL) receptor or apolipoprotein B (apoB). We could exclude the third FH locus as a cause of FH by genetic linkage analysis in the families taken together. Since haplotype analysis of each family nevertheless suggested that the FH phenotype co-segregated in a manner consistent with linkage to the third FH locus in three small pedigrees, we performed sequencing analysis without being able to demonstrate mutations in the proprotein convertase subtilisin/kexin type 9 (PCSK9) gene, the main candidate gene in the third FH locus. By the same combination of genetic linkage and molecular analysis we could also exclude mutations in the gene for the LDL receptor adaptor protein and in the gene for cholesterol-7-alpha-hydroxylase as causes of FH in our sample. Although not indicating linkage to any known loci, our data still indicate that another dominant gene may be involved in causing a FH phenotype. PMID- 15530919 TI - Carotid intima-media thickness, cardiovascular risk factors and albuminuria in a remote Australian Aboriginal community. AB - BACKGROUND: Rates of cardiovascular disease and renal disease in Australian Aboriginal communities are high, as is the prevalence of some 'traditional' cardiovascular (CV) risk factors, such as diabetes and cigarette smoking. Recent work has highlighted the importance of markers of inflammation, such as C reactive protein (CRP), homocysteine and albuminuria as predictors of cardiovascular risk in urban westernized settings. It is not clear how these factors relate to outcome in the setting of these remote communities, but very high CRP concentrations have been shown in this and other Aboriginal communities. METHODS AND RESULTS: In a cross-sectional survey including 237 adults in a remote Aboriginal community in the Northern Territory of Australia, we measured carotid intima-media thickness (IMT), together with blood pressure, diabetes, lipid levels, smoking and albuminuria, CRP and fibrinogen, serum homocysteine concentration, and IgG titres for Chlamydia pneumoniae, Helicobacter pylori and cytomegalovirus. Median carotid IMT was 0.63 [interquartile range 0.54-0.71] mm. As a categorical outcome, the prevalence of the highest IMT quartile ('increased IMT', > or =0.72 mm) was compared with the lower three quartiles. Increased IMT was associated in univariate analyses with greater waist circumference, systolic BP, fibrinogen and serum albumin concentrations, urine albumin/creatinine ratio and older age as continuous variables. Associations of increased IMT with some continuous variables were not linear; univariate associations were seen with the highest quartile (versus all other quartiles) of CRP and homocysteine concentration and CMV IgG titre. In a multivariate model age, smoking, waist circumference and the highest quartile of CRP concentrations (> or =14 mg/l) remained significant predictors of IMT > or =0.72 mm. CONCLUSIONS: Measurement of carotid IMT was possible in this remote setting. Increased IMT (> or =0.72 mm) was associated with increased CRP concentrations over a range that suggests infection/inflammation may be important determinants of cardiovascular risk in this setting. The associations of IMT with markers of renal disease seen in univariate analyses were explained in this analysis by confounding due to the associations of urine ACR with other risk factors. PMID- 15530920 TI - Effects of statin therapy on the progression of carotid atherosclerosis: a systematic review and meta-analysis. AB - Observational studies suggested that the progression of carotid atherosclerosis and plaques might be an independent risk factor for coronary artery disease (CAD) and stroke, prior researches of different dosage of statins therapy on the progression of carotid atherosclerosis in patients had small sample sizes and different the results. We aimed to evaluate the efficacy and safety of statins therapy on the rate of carotid atherosclerosis progression through randomized controlled trials (RCTs). We searched MEDLINE (1980-September 2003), the Cochrane Controlled Trials Register, EMBASE (1985-September 2003), Science Citation Index and PUBMED (updated through December 2003). Additional articles were obtained from reference lists of relevant reviews and papers in manual searches. Ten studies (eight studies between statins and placebo, two studies between aggressive statins and conventional therapy) were included. Two reviewers extracted abstracts, and appraised the Jadad Score for Grading Quality of Randomized Controlled Trials independently, and discrepancies were resolved by consensus with a third reviewer. We analyzed the 10 trials with 3443 individuals age range from 30 to 70 years old and follow-up for 1-4 years. Eight studies showed that the rates of carotid IMT progression were consistently significantly different between statins and placebo, the Z score for overall effect of IMT was 10.41 (P<0.00001), Total 95%CI of weighted mean difference (WMD) between two groups was -0.02235 [-0.02656, -0.01614] (mm/y), the chi(2)-test for heterogeneity of IMT between eight studies was 685.33 (P<0.00001), two studies between aggressive statins and conventional statins demonstrated that the Z score for overall effect of IMT was 15.85 (P<0.00001), total 95%CI of WMD between two groups was -0.06326 [-0.07108, -0.05544], chi(2)-test for heterogeneity of IMT between two studies was 3.75 (P=0.05). Conventional statins were not significantly different of adverse events and withdrawal than placebo (P>0.05). Our findings suggest that conventional statins therapy are efficient and safe to decrease the rate of carotid atherosclerosis progression in a long-term, and aggressive statins may provide superior efficacy for carotid atherosclerosis regression. PMID- 15530921 TI - Locating Ath8, a locus for murine atherosclerosis susceptibility and testing several of its candidate genes in mice and humans. AB - A previous study revealed that the difference in susceptibility to atherosclerotic lesions between inbred mouse strains SM/J and NZB/BlNJ was determined by one major locus (Ath8). In this study a (SM/J x NZB/BlNJ) F(1) x SM/J backcross localized Ath8 by quantitative trait locus mapping to chromosome 4 with a suggestive LOD score of 2.7. This quantitative trait locus (QTL) was confirmed using an (SM/J x NZB/BlNJ) intercross; Ath8 mapped to a 23cM region with a significant LOD score of 3.6. The genes for toll-like receptor 4 (T1r4), arachidonic acid epoxygenase (Cyp2j5), and angiopoietin-like protein 3 (Angptl3) map to this region. These candidate genes were analyzed for expression and sequence differences in the mouse and for associations with cardiovascular traits in human. Sequence analysis of Angptl3 shows a base pair substitution in SM, the susceptible strain, giving rise to an amino acid change in the fibrinogen homology domain of the protein. We found a significant association between ANGPTL3 and atherosclerotic lesions (P < 0.05) in human. These results suggest that Angptl3 is involved in atherosclerosis susceptibility in both mouse and human. PMID- 15530922 TI - On-road measures of pedestrians' estimates of their own nighttime conspicuity. AB - PROBLEM: Most pedestrian fatalities occur at night. Although researchers have long understood that drivers have difficulty seeing pedestrians at night and that reflective clothing can dramatically enhance pedestrian conspicuity, the extent to which pedestrians understand these facts is unclear. This experiment quantified pedestrians' estimates of the ability of an approaching driver to recognize the presence of roadside pedestrians. METHOD: Ten younger and 10 older participants walked in place on the far shoulder of a closed-road circuit and pressed a button when they were confident that the approaching driver could first recognize that a pedestrian was present. RESULTS: Pedestrians overestimated their visibility and dramatically underestimated the benefit of conspicuity treatments. CONCLUSIONS: These results suggest that pedestrians fail to understand the magnitude of the nighttime conspicuity problem and the value of conspicuity treatments. Pedestrians may therefore unknowingly place themselves in danger at night. IMPACT: These results underscore the need to educate pedestrians about the dangers of interacting with traffic at night and about treatments that increase their safety. PMID- 15530923 TI - The Swedish driver licensure examination: exploration of a two-stage model. AB - INTRODUCTION: Sweden, together with several countries in the world, has no compulsory educational requirement prior to the drivers license examination. Having a valid license examination (i.e., a knowledge test and a driving test) is therefore important in order to ensure competent drivers. METHOD: The aim of this paper is to explore the possibility of a two stage testing approach for the Swedish licensure examination. In order to explore the relationship between the knowledge test and the road test, a descriptive and a logistic regression approach were carried out. RESULTS: The result indicates that how the driving candidates perform on the road test depends on their performance on the knowledge test. Therefore, the Swedish drivers licensing examination is suitable for a two stage testing model. PMID- 15530924 TI - Exploratory analysis of the safety climate and safety behavior relationship. AB - PROBLEM: Safety climate refers to the degree to which employees believe true priority is given to organizational safety performance, and its measurement is thought to provide an "early warning" of potential safety system failure(s). However, researchers have struggled over the last 25 years to find empirical evidence to demonstrate actual links between safety climate and safety performance. METHOD: A safety climate measure was distributed to manufacturing employees at the beginning of a behavioral safety initiative and redistributed one year later. RESULTS: Multiple regression analysis demonstrated that perceptions of the importance of safety training were predictive of actual levels of safety behavior. The results also demonstrate that the magnitude of change in perceptual safety climate scores will not necessarily match actual changes (r=0.56, n.s.) in employee's safety behavior. DISCUSSION: This study obtained empirical links between safety climate scores and actual safety behavior. Confirming and contradicting findings within the extant safety climate literature, the results strongly suggest that the hypothesized climate-behavior accident path is not as clear cut as commonly assumed. SUMMARY: A statistical link between safety climate perceptions and safety behavior will be obtained when sufficient behavioral data is collected. IMPACT ON INDUSTRY: The study further supports the use of safety climate measures as useful diagnostic tools in ascertaining employee's perceptions of the way that safety is being operationalized. PMID- 15530925 TI - Have young workers more injuries than older ones? An international literature review. AB - PROBLEM: Two questions were posed in this global literature review: Do young workers have a higher occupational injury rate? Are the injuries of young workers more often fatal than those of older workers? METHOD: The studies of nonfatal and fatal injuries were collected based on the following criteria: (a) published in peer-reviewed journals; (b) the young workers were under 25 years of age; (c) the injury rate or fatality rate of young workers and the overall rate was published; and (d) description of the population and the number of injuries was presented. RESULTS: The majority of 63 nonfatal studies reported showed that young workers had a higher injury rate than older workers. Twenty-nine out of 45 studies on fatal occupational injuries indicated that young workers had a lower fatality rate than older workers. These results are clearer for men than for women. IMPACT ON INDUSTRY: The results showed that young men were a risk group for occupational injuries. However, the injuries of young workers were reported as less often fatal than those of older workers. PMID- 15530926 TI - Evaluation of California's graduated driver licensing program. AB - PROBLEM: California's graduated driver licensing (GDL) program was implemented to reduce the high crash risk of teenage drivers. The program enhancements made in 1998 were evaluated in this study using methods that rule out the noticeable downward trend in California teen's fatal/injury crash rates as a possible explanation for any observed crash reductions that could otherwise be mistaken for program effects. METHOD: Monthly per capita fatal/injury crash rates for 15 to-17-year-olds and proportional representations of 16-17-year-olds' nighttime and passenger crashes were analyzed using intervention time series analysis. RESULTS: After removing trend, seasonality, and transition effects in the data, no overall reductions in fatal/injury crashes for 15-17-year-olds or 16-year-olds (separately) were found to be associated with the 1998 program enhancements, suggesting no overall exposure reduction effect of the longer instruction permit period nor increased competency associated with the higher number of required practice hours. However, the 12-month nighttime and 6-month passenger restrictions were found to be associated with annual savings of 55 and 816 fatal/injury crashes, respectively. DISCUSSION: Finding no overall impact of the 1998 GDL enhancements was not surprising given findings of an earlier survey suggesting that California teens and parents were largely already practicing program requirements prior to implementation and were not fully complying with the program requirements afterwards. Though the observed crash savings associated with the restrictions were of modest size, this is the first study to evidence a direct positive impact of the passenger restriction component of GDL programs. Larger reductions could be realized if the nighttime restriction started earlier and parents/law enforcement could be motivated to better enforce these restrictions. IMPACT ON INDUSTRY: The findings provide support for passenger and nighttime restriction components of GDL programs. PMID- 15530927 TI - High visibility safety apparel and nighttime conspicuity of pedestrians in work zones. AB - PROBLEM: Every year numerous occupational fatalities result from pedestrians being struck by motor vehicles intruding into work zones. METHOD: Attributes of retroreflective personal safety garments on pedestrian conspicuity at night were assessed in a field study. Using instrumented vehicles on a closed track, participants drove through simulated work zones attempting to detect pedestrians located in the work zones. RESULTS: Configuration of the retroreflective trim, trim color, placement in the work zone, and driver age significantly affected pedestrian conspicuity. Intensity and the amount of retroreflective trim did not. DISCUSSION: Personal safety garments incorporating retroreflective trim significantly improve pedestrian conspicuity in work zones. SUMMARY: The results emphasize the importance of retroreflective trim on personal safety garments, particularly if the trim is located on garment sleeves. IMPACT ON INDUSTRY: We examine the design attributes that contribute to making a personal safety garment conspicuous. The results have implications regarding preferred garment designs, industry standards, and service life of personal safety garments. PMID- 15530928 TI - An in-depth look at parent-imposed driving rules: their strengths and weaknesses. AB - INTRODUCTION: With a growing interest in increasing parental involvement in teen driving, it is important to find out what parents are already doing. METHOD: This study assessed the content, delivery, rigidity, and consequences of 143 driving rules reported by 24 parent-teen dyads. RESULTS: Strengths included that driving rules covered the full range of concerns, especially night driving limits and passenger limits, and most parents and teens reported that violations would be followed by consequences, especially talk/warn or no driving. Weaknesses included that many rules were not very strict and only half showed parent-teen agreement on content. CONCLUSIONS: These findings suggest that teen driving rules are not clearly defined. PMID- 15530930 TI - Nonfatal motor-vehicle animal crash-related injuries -- United States, 2001-2002. PMID- 15530929 TI - Validation of the propensity for angry driving scale. AB - PROBLEM: This study examined the validity of the Propensity for Angry Driving Scale (PADS). Measuring road rage: Development of the Propensity for Angry Driving Scale in predicting aggressive driving. METHOD: The PADS and the Driving Anger Scale. Development of a driving anger scale. Psychological reports, 74, 83 91.) were administered to 232 college student volunteers with measures of aggressive and risky driving. RESULTS: Convergent and discriminant validity of the PADS were supported through relationships among measures of similar constructs. The PADS significantly (p<.05) predicted moving tickets, minor accidents, aggressive driving, risky driving, and maladaptive driving anger expression, above and beyond gender, miles driven per week, and trait anger. DISCUSSION: Findings suggest that the PADS is a useful predictor of aggressive driving and has some advantages over the DAS. IMPACT ON INDUSTRY: The PADS is an effective predictor of aggressive driving that complements established measures like the DAS and provides researchers with another valuable tool for the assessment of aggressive driving. PMID- 15530931 TI - Plus ca change, plus ca meme chose! PMID- 15530932 TI - Celebrating Public Health Lives: public health and the Seventh Earl of Shaftesbury. PMID- 15530933 TI - Evaluating health impact assessment. AB - Health impact assessment (HIA) is a developing approach that assesses the health impacts of a proposal on a population, and produces a practical set of recommendations to inform the decision-making process of the proposal. The purpose is to influence decision makers to increase positive health impacts of a proposal and decrease negative impacts. Most work within the HIA field to date has focused on methodological development and actually carrying out HIAs. Little attention has been paid to the formal evaluation of the HIA approach and whether or not HIA works (if and how the HIA approach informs the decision-making process and, in particular, if it contributes to improving health and reducing inequalities). With the drive towards evidence-informed policy, HIA also needs to show whether it adds value to the decision-making process, given the significant resources often involved in carrying out an HIA. A suggested approach to evaluating HIAs is presented, as well as its relationship with monitoring, drawing on the significant public health evaluation literature that already exists. Methodologies appropriate for use, and examples of indicators suitable for HIA evaluation, are provided. We suggest that typical HIA evaluations should focus on the process of the HIA and the impact that it has on the decision-making process, rather than attempting to evaluate long-term health outcomes or whether predicted impacts actually occurred. PMID- 15530934 TI - Trends in socio-economic differentials in cigarette smoking behaviour between 1990 and 1998: a large prospective study in Korean men. AB - OBJECTIVES: To investigate the magnitude and time trends in socio-economic differentials in cigarette smoking behaviour. STUDY DESIGNS: A prospective observational study of male civil servants followed-up for 8 years. SETTING: All civil service offices in Korea. PARTICIPANTS: A total of 322,991 male civil servants aged 30-49 years on whom information on monthly salary was available and who had reported cigarette smoking behaviour between 1990 and 1998. RESULTS: Smoking prevalence was 60.1% in 1990 and was inversely associated with salary level. The difference in smoking prevalence between the highest and the lowest salary groups gradually widened from 13.0% in 1990 to 16.1% in 1998. The relative index of inequality (RII) as a measure of socio-economic inequality in smoking behaviour was 1.91 [95% confidence interval (CI) 1.86-1.97] in 1990 and increased between 1990 and 1998. The RII for smoking cessation was 0.60 in 1990 (95% CI 0.58-0.62) and gradually decreased as time lapsed. CONCLUSIONS: An increasing trend in socio-economic inequality in smoking behaviour was observed in this study. PMID- 15530935 TI - Arsenicosis in Bangladesh: prevalence and socio-economic correlates. AB - The potential effects of arsenic-contaminated drinking water on health are of concern, but our understanding of the risk factors of arsenicosis remains limited. This study assessed the prevalence of and socio-economic differentials in arsenic-associated skin lesions in a rural community in Bangladesh. Data were collected from a village where the Bangladesh Rural Advancement Committee has operated a health surveillance system and a community-based arsenic mitigation project since 1999. In total, 1654 residents in the study village were examined in May 2000 for arsenic-associated lesions on their skin. Socio-economic information was extracted from the surveillance system database covering the village. Nearly 2.9% of the study population had clinical manifestations of arsenic poisoning. The prevalence of arsenicosis was associated with age, sex, education and the economic status of the household. Multivariate analysis identified age and economic status as significant predictors of arsenicosis controlling for education and gender. In conclusion, a clear understanding of the socio-economic distribution of arsenicosis in different demographic and socio economic groups will be useful in identifying the high-risk groups from arsenic affected communities. More studies are needed to design effective interventions to mitigate the effects of arsenic in Bangladesh. PMID- 15530936 TI - Effect of several sociodemographic factors on measles immunization in children of Eastern Turkey. AB - This study was performed to determine the status of measles immunization and the effect of several sociodemographic factors on immunization in children aged between 10 months and 6 years. Using cluster sampling, 663 healthy children were selected at random from three provinces in Eastern Turkey. The immunization histories of these children were obtained from their immunization cards. With respect to their vaccination status, children were categorized as vaccinated, unvaccinated or unknown. If the child had no immunization card, he or she was classified as unknown. Of these children, 81.6% were vaccinated and 15.1% were unvaccinated. The vaccination status was not known in 3.3% of the children. In children aged 10-12 months, the vaccination rate was 68.6%. For age groups of 1 2, 2-3, 3-4, 4-5 and 5-6 years, the vaccination rates were 84.2, 82.2, 85.3, 82.1 and 76.8%, respectively, but these differences were not significant. The vaccination rates increased in parallel with maternal education level (P = 0.009). Also, vaccination rates were significantly correlated with settlement area (P = 0.036), and were higher in urban regions than suburban and rural regions. There was no difference in vaccination rates with respect to gender, paternal education level, number of siblings and socio-economic status. The results of this study show that the level of immunization necessary for measles elimination has not yet been reached in Eastern Turkey. Priority should be given to increase the immunization levels to 90-95% among children. PMID- 15530937 TI - Penetrating eye injuries from southeastern Anatolia region of Turkey. AB - OBJECTIVES: This study analysed 420 patients with penetrating eye injuries who presented for treatment at the Eye Clinic of Dicle University Hospital in Turkey. The aim of the study was to identify preventable risk factors. METHODS: A retrospective evaluation of patients who presented between January 1995 and December 2000 was undertaken. Cases were examined with regard to age, sex, profession, which eye was traumatized, reason for the trauma, nature of the trauma, time between injury and operation, and accompanying pathologies. RESULTS: This study found that most penetrating eye injuries occurred in children aged 0 15 years, and these were mainly caused by accidents during play. For patients aged 16-60 years, penetrating eye injuries were mainly caused by work-related accidents. CONCLUSION: Most penetrating eye injuries were caused by preventable accidents. Many accidents and their resulting injuries could have been prevented by education, greater intensive care and workplace safety. PMID- 15530938 TI - Do rates of hospital admission for falls and hip fracture in elderly people vary by socio-economic status? AB - OBJECTIVE: To determine the relationship between hospital admissions for falls and hip fracture in elderly people and area characteristics such as socio economic deprivation. STUDY DESIGN: Ecological study of routinely collected hospital admissions data for falls and hip fracture in people aged 75 years or over for 1992-1997, linked at electoral ward level with characteristics from census data. METHODS: In total, 42,293 and 17,390 admissions were identified for falls and hip fracture, respectively, from 858 electoral wards in Trent. Rate ratios (RRs) for hospital admissions for falls and hip fracture were calculated by the electoral wards' Townsend score divided by quintiles. RRs were estimated by negative binomial regression and adjusted for the ward characteristics of age, gender, ethnicity, rurality, proportion of elderly people living alone and distance from hospital. RESULTS: There was a small but statistically significant association at electoral ward level between hospital admissions for falls and the Townsend score, with the most deprived wards having a 10% higher admission rate for falls compared with the most affluent wards (adjusted RR 1.10, 95% CI 1.01 1.19). No association was found between hospital admission for hip fracture and deprivation (adjusted RR 1.05, 95% CI 0.95-1.16). CONCLUSION: There is some evidence of an association at electoral ward level between hospital admissions for falls and socio-economic deprivation, with higher rates in deprived areas. No such association was found for hip fracture. Further work is required to assess the impact of interventions on reducing inequalities in hospital admission rates for falls in elderly people. PMID- 15530939 TI - Factors associated with influenza vaccination among elderly Spanish women. AB - OBJECTIVES: Influenza vaccination is an effective intervention to diminish morbidity and mortality associated with this disease in aged populations and at risk groups. The objective of this work was to describe population patterns of vaccination among Galician women and to identify factors associated with vaccination. STUDY DESIGN: Cross-sectional study. METHODS: Cases aged 65 years and over were selected from the Women's Social and Health Interview, Galicia 2000 (n = 1111). The association between influenza vaccination last season and several sociodemographic, lifestyle, health status and health services variables was assessed by logistic regression. RESULTS: In total, 56.3% of cases had received the influenza vaccine. The following variables were significantly associated with vaccination: age 70-74 years (odds ratios, OR=1.56; 95% CI: 1.09-2.26); age> or =75 years (OR=1.88; 95% CI: 1.31-2.71); residence in towns with 5000-20,000 inhabitants (OR=1.79; 95% CI: 1.16-2.77); annual income 6.010 (OR=1.39; 95% CI: 1.01-1.90); unfavourable self-perception of health (OR=1.46; 95% CI: 1.06-2.00); not being a caregiver (OR=1.67; 95% CI: 1.17-2.38); married (OR=1.45; 95% CI: 1.05-2.01); tetanus vaccination (OR=1.43; 95% CI: 1.07-1.93); and visiting a physician in the last 2 years (OR=4.83; 95% CI: 2.61-8.93). CONCLUSIONS: The level of vaccination among Galician women is low, although it is higher than that in Spanish women overall. This work has identified groups of women who are less likely to be vaccinated, and who should be targeted in future vaccination campaigns. PMID- 15530940 TI - Psychosocial correlates of substance use among adolescents in Mersin, Turkey. AB - The objective of this study was to determine the effects of psychosocial factors such as peer group, family and academic self-perception on smoking, alcohol and substance use by adolescents living in Mersin, Turkey. The study included a total of 3282 students from the sixth and tenth grades and college. The number of participating students required from each school was obtained through stratification, and by weighing the enrolled student population in each subgroup. The final sample was derived using a simple random sampling technique. A 45-item self-administered questionnaire was used. The questionnaire included questions about socio-demographic characteristics and lifetime and current (i.e. within the past month) use of cigarettes, alcohol, cannabis, inhalants and other illicit drugs (heroin, cocaine, sedative-hypnotic drugs, etc.). This study found that: (1) higher socio-economic status of the family increased the likelihood of smoking and alcohol use in adolescents; (2) the prevalence of alcohol use was higher in adolescents whose mothers had a higher educational level and whose mothers and fathers drank alcohol; (3) there was a significant association between substance use and having a peer who used a substance; and (4) the prevalence of smoking was significantly higher in students who perceived their academic performance to be poor. Understanding the role and importance of psychosocial factors associated with smoking, alcohol and substance use will be crucial to develop preventive measures for adolescents. PMID- 15530941 TI - Treatment of hypertension in Spanish primary care centres: is it evidence based? PMID- 15530942 TI - An analysis of the Cuban health system. PMID- 15530943 TI - Dysmorphology and the orbital region: a practical clinical approach. AB - Dysmorphology is the field of medicine focusing on congenital developmental abnormalities due to exogenous teratogens, chromosomal anomalies, or to a defect in a single gene. Numerous syndromes have been reported and a growing number of genes or chromosomal anomalies are identified. The clinical observation of the face remains an essential part of the clinical evaluation of the patients. The orbital region, as other regions of the face, should be systematically evaluated. Orbital malformations can be isolated or part of a syndrome. In the diagnostic process, the orbital anomaly can be classified as a major feature (essential for the diagnosis), a moderate feature (important but not essential for the diagnosis), or a minor feature (contributing weakly to the diagnosis). The diagnoses of the main orbital anomalies in dysmorphology are reviewed and illustrated with relevant examples of syndromes that are presented as well as the usual landmarks used in clinical practice. Abnormal position of the eyes in syndromes such as hypertelorism, hypotelorism, primary or secondary telecanthus, asymmetry, and proptosis are discussed. Eyelid anomalies, such as cryptophthalmos, ablepharon, blepharophimosis, euryblepharon, or anomalies at the level of the eyelashes and eyebrows are described. PMID- 15530944 TI - Listing's law: clinical significance and implications for neural control. AB - Listing's law governs the three-dimensional orientation of the eye and its axes of rotation. It states that, when the head is fixed, there is an eye position called primary position, such that the eye assumes only those orientations that can be reached from primary position by a single rotation about an axis in a plane called Listing's plane. Listing's law can also be expressed in terms of any initial eye position, not just primary position (Listing's half-angle rule). The binocular extension of Listing's law is equivalent to Listing's law when the vergence angle is zero, and adjusts the eyes' torsion when they converge. Listing's law holds during fixation, saccades, smooth pursuit, and vergence, but not during sleep and vestibulo-ocular reflex, suggesting that it is actively implemented by a neural mechanism. Orbital constraints, such as "pulleys," may also play a role. Adherence to Listing's law and its extension may serve the purpose of optimizing motor efficiency, or simplifying neural processing for binocular vision, or both. The study of Listing's law not only allows us to understand the organization of neural and mechanical factors in the control of three-dimensional eye movements, it also has important clinical implications for strabismus and the optimal management of this disorder. PMID- 15530945 TI - Evolution, techniques, clinical outcomes, and pathophysiology of LASEK: review of the literature. AB - Laser subepithelial keratomileusis (LASEK) is a relatively new laser surgical procedure that combines certain elements of both laser in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK) to improve the risk/benefit ratio. Diluted alcohol is used to loosen the epithelial adhesion to the corneal stroma. The loosened epithelium is moved aside from the treatment zone as a hinged sheet. Laser ablation of the subepithelial stroma is performed before the epithelial sheet is returned to its original position. We reviewed the literature regarding modifications of the technique, indications, outcomes, and complications, as well as wound healing after LASEK surgery. This literature review of 1,421 LASEK treated eyes provided many findings: 1) The long-term stable results in the absence of serious complications, such as infections, recurrent erosions, scars, or late-onset corneal haze formation in patients re-examined up to 5 years after LASEK; 2) Epithelial closure with recovery of functional vision was completed at days 4 to 7 in most cases; 3) A tendency toward overcorrection with PRK nomograms; 4) We hypothesize that this tendency may be due to the decreased wound healing response, which may lead to myopic regression in PRK; and 5) Postoperative discomfort and prolonged visual recovery until the epithelium closes remain the biggest disadvantages of LASEK compared to LASIK. LASEK surgery is especially valuable in patients with thin corneas who would not qualify for LASIK surgery. However, a potential superiority of LASEK to LASIK in wavefront guided ablations still remains speculative. PMID- 15530946 TI - A case of biopsy-negative temporal arteritis--diagnostic challenges. AB - A patient with systemic symptoms but no visual loss was investigated for suspected giant cell arteritis. Initial temporal artery biopsy was reported as negative; however, she returned with visual loss 2 months later, and the diagnosis of giant cell arteritis was confirmed with a subsequent biopsy. In hindsight, signs suggestive of the disease were present in the original biopsy, although the usual diagnostic features were absent. PMID- 15530947 TI - Juvenile xanthogranuloma of the corneoscleral limbus: case report and review of the literature. AB - Juvenile xanthogranuloma is a benign histiocytic cutaneous disorder mainly occurring in infants which may affect the eye. Ocular manifestations usually present in the form of iris lesions with secondary hyphema and glaucoma, but juvenile xanthogranuloma may also present as a corneoscleral limbal mass. We present the case of an 18-month-old female infant with ocular juvenile xanthogranuloma occurring as a corneoscleral limbal mass without associated cutaneous or systemic findings. The limbal mass lesion was excised by lamellar dissection and histopathological studies revealed histiocytes admixed with lymphocytes, plasma cells, and eosinophils; foreign body and Touton giant cells present throughout the specimen confirmed the diagnosis of juvenile xanthogranuloma. At 14-month follow-up, there was no recurrence of the limbal mass. The natural history of this uncommon ocular condition is discussed and the relevant literature reviewed. Juvenile xanthogranuloma should be considered in the differential diagnosis of any corneoscleral limbal mass lesion, particularly in children. PMID- 15530948 TI - Challenges in ocular infectious diseases and the evolution of anti-infective therapy. PMID- 15530949 TI - Transsphenoidal diplopia. PMID- 15530951 TI - Anthocyanosides of Vaccinium myrtillus (bilberry) for night vision--a systematic review of placebo-controlled trials. PMID- 15530953 TI - Ocular tuberculosis masquerading as ocular tumors. PMID- 15530955 TI - Polypoidal choroidal vasculopathy. PMID- 15530957 TI - A cost-effectiveness comparison of bimatoprost versus latanoprost in patients with glaucoma or ocular hypertension. PMID- 15530960 TI - Biologically active substances from amphibians: preliminary studies on anurans from twenty-one genera of Thailand. AB - Amphibian skin has been the source of a wide variety of biologically active substances, but less than one-third of the known genera of amphibians have been probed for such active substances. Skins of 21 genera of anurans from Thailand have now been investigated for noxious secretions, toxic substances, and alkaloids. Four genera of bufonid toads (Bufo, Ansonia, Leptophryne, Pedostipes) were toxic due to the presence of bufadienolides or bufadienolide-like compounds. Two species of ranid frogs (Rana raniceps, Rana signata) were toxic, perhaps due to the presence of toxic peptide(s). Two species of rhacophorid frogs (Polypedates) were slightly noxious/toxic. One species of microhylid frog (Kaloula pulchra) was noxious. Trace amounts of pumiliotoxin alkaloids were detected in a ranid frog (Limnonectes kuhli). A further 18 species did not exhibit noxious or toxic properties to a significant extent. PMID- 15530961 TI - The case for verapamil use in alarming jellyfish stings remains. PMID- 15530963 TI - Subchronic oral toxicity of microcystin in common carp (Cyprinus carpio L.) exposed to Microcystis under laboratory conditions. AB - The subchronic oral toxicity of microcystin in common carp (Cyprinus carpio L.) was investigated in this study. The fish (mean body weight of 322+/-36 g, n=10) were orally exposed to Microcystis by feeding with bloom scum at a dose of 50 microg microcystins/kg body weight under laboratory conditions for 28 days. Growth assay results showed that microcystin could completely inhibit the growth of carp, but failed to change the fish hepatosomatic index. Ultrastructural examination by electron microscope revealed severe damage in hepatocytes derived from the treated fish. Serum biochemical assays with commercial kits indicated that alanine aminotransferase and aspartate aminotransferase activities were significantly increased as compared to control levels, but gamma-glutamyl transferase, alkaline phosphatase and lactate dehydrogenase activities remained unchanged. Protein phosphatase inhibition assay revealed that the microcystin concentrations were 261.0+/-108.3 ng microcystin-LR equivalent/g fresh weight in hepatopancreas and 38.3+/-12.3 ng microcystin-LR equivalent/g fresh weight in muscle. The latter is above the limit recommended by the World Health Organization for human consumption. Therefore, we recommend that a warning system be instituted for announcing the occurrence of microcystin-producing water bloom and the possible risk of human intoxication. PMID- 15530964 TI - Larvicidal action of ethanolic extracts from fruit endocarps of Melia azedarach and Azadirachta indica against the dengue mosquito Aedes aegypti. AB - Ethanolic extracts from the kernels of ripe fruits from the Indian Lilac Melia azedarach and from the well-known Neem tree, Azadirachta indica were assayed against larvae of Aedes aegypti, the mosquito vector of dengue fever. The lethality bioassays were carried out according to the recommendations of the World Health Organization. Extracts were tested at doses ranging from 0.0033 to 0.05 g% in an aqueous medium for 24 and 48 h, at 25 or 30 degrees C, with or without feeding of the larvae. LC50, LC95 and LC99 were determined. Both seed extracts proved lethal for third to fourth instar larvae. Non-fed A. aegypti larvae were more susceptible to Azadirachta extracts at both temperatures. Under a more realistic environmental situation, namely with fed larvae at 25 degrees C, the death rates caused by the Melia extract were higher, although at 30 degrees C the extract of Azadirachta had an even higher lethality. Inter allia, the LC50 values for the crude extracts of these two members of the Meliaceae ranged from 0.017 to 0.034 g% while the LC99 values ranged from 0.133 to 0.189 g%. Since no downstream processing was undertaken to purify the active agents in the extracts, our findings seem very promising, suggesting that it may be possible to increase the larvicidal activity further by improving the extraction and the fractionation of the crude limonoids, for instance removing the co-extracted natural fats. PMID- 15530965 TI - The efficacy of two antivenoms against the in vitro myotoxic effects of black snake (Pseudechis) venoms in the chick biventer cervicis nerve-muscle preparation. AB - Neurotoxicity is rarely seen following human systemic envenoming by Australasian black snakes (genus Pseudechis) with myotoxicity being the most prominent feature following bites by some species. This study investigated the in vitro myotoxicity of venoms from seven species of Australasian Pseudechis and determined the efficacy of CSL black and tiger snake antivenoms. All Pseudechis venoms (10 microg/ml) significantly inhibited direct twitches of the chick biventer cervicis nerve-muscle preparation (p<0.05, one-way ANOVA). Prior addition of black snake antivenom (5 U/ml) prevented the inhibitory effects of all Pseudechis venoms (p<0.05, one-way ANOVA), except Pseudechis butleri. Prior addition of tiger snake antivenom (5 U/ml) prevented the venom-induced reduction in direct twitches to Notechis scutatus venom and all Pseudechis venoms (p<0.05, one-way ANOVA), with the exception of Pseudechis australis and Pseudechis colletti venoms. Black or tiger snake antivenom (5 U/ml) added 1 h after the addition of venom inhibited further reduction of direct twitches by N. scutatus and most Pseudechis venoms, but did not significantly restore twitch height. PLA2 activity was found in all venoms with the following rank order: Pseudechis porphyriacus>P. australis>Pseudechis papuanus>P. butleri>Pseudechis guttatus> or =Pseudechis pailsii>P. colletti>N. scutatus. The results of the present study suggest that Australasian Pseudechis venoms possess variable myotoxic activity. The ability of black or tiger snake antivenom to prevent or inhibit further venom-induced effects varied across the genus. PMID- 15530966 TI - Insights into the loss of muscle mass following B. jararacussu venom in mice. AB - Bothrops jararacussu snake venom produces myonecrosis and nerve degeneration. In this work, we investigated whether nerve lesions or impaired muscle regeneration contributed to the permanent loss of muscle mass, a long-term sequela of envenoming. The right soleus muscle of adult male mice was injected with B. jararacussu venom (80 microg) while the left muscle received only saline (control). The mice were killed after 2 and 3 months and the muscles were removed and processed for examination by transmission electron microscopy and light microscopy. The nerve fibers, Schwann cells and neuromuscular junctions had regenerated in venom-treated muscle. The total number of muscle fibers was significantly lower (p<0.05) than in the control (617+/-48 versus 1235+/-97, respectively; mean+/-SEM, n=10). These results show that the loss of muscle mass was most likely related to a decrease in the ability of the muscle to regenerate rather than to nerve lesions. PMID- 15530967 TI - Histopathological changes and inflammatory response induced by Tityus discrepans scorpion venom in rams. AB - Anesthetized rams envenomed s.c. with 40 microg/kg Tityus discrepans scorpion venom developed fasciculation, hypothermia, polyuria, pulmonary wet rales, tachypnea, respiratory distress and arrhythmia. Rams developed a cascade of inflammation reactions, characterized by activation of macrophages, fibroblasts and neutrophils, neutrophil infiltration and aggregation, vasculitis, arteritis and abundant fibrin deposition. At the inoculation site, venom was detected by immunohistochemistry in the extra cellular matrix, lymphatic vessels' and venules' lumen, inside macrophages and surrounding nerves. Extra cellular matrix was degraded at the inoculation site perhaps by activated neutrophils. Envenoming produced hepatocytes with Mallory body-like vacuoles which may be due to the increased plasmatic levels of TNF-alpha and IL6. Venom produced degranulation and vacuolization of acinary cells as well as interstitial swelling and necrosis. Necrosis of the Langerhan's islets occurred occasionally. Lungs showed the most deleterious effects developing wall collapse and necrosis, diffuse injury of the alveolar capillary barrier, interstitial and alveolar fibrin deposits with strong neutrophil infiltration. Massive infiltration of lymphocytes and macrophage occurred in the intestinal submucose, to the point that it modified villi and intestinal folding morphology. Envenomation developed a marked leukocyte aggregation surrounding nerves at the inoculation site. This study reveals that beyond its neurotoxicity, Tityus venom produces a severe and widespread inflammatory syndrome, expressed as histopathological changes at the site of inoculation, as well as in remote organs such as pancreas, lungs, intestine and liver. Our results suggest that not all remote targets are directly affected by the venom but that, as proposed earlier, are modified by inflammation by products produced elsewhere. PMID- 15530968 TI - Jararhagin, a snake venom metalloproteinase-disintegrin, stimulates epithelial cell migration in an in vitro restitution model. AB - The snake venom metalloproteinase-disintegrin jararhagin (JG) has no chemotactic activity but stimulates the migration of neutrophils in vivo through a mechanism still unclear. In this study we investigated the effects of jararhagin on epithelial cell adhesion and migration in vitro. F-actin arrangement and the distribution of laminin, fibronectin, several integrins and phosphorylated Focal Adhesion Kinase (FAK) were studied using rhodamine-phalloidin and immunofluorescence. Maximum stimulation of migration (about 100%) was obtained with 5 microg/ml JG, with about 38% inhibition of cellular adhesion. In migratory cells the toxin stimulated the formation of filopodia, lamellipodia and stress fibers. The pericellular fibronectin matrix was lost in migrating cells, while laminin was less affected. The toxin stimulated FAK phosphorylation and the recruitment of alphav-containing integrins to focal contacts, whereas integrins containing the alpha2 subunit were reduced in these junctions. Inactivation of the toxin with 1,10 phenanthroline showed that the catalytic activity is important for the effect of jararhagin on cell migration, FAK phosphorylation and for the recruitment of alphav, but not as much for the anti-adhesive effect. In conclusion, jararhagin stimulates the migration of epithelial cells in vitro through a mechanism that involves its proteolytic activity, qualitative changes in cellular adhesion and the formation of actin-rich cellular processes. PMID- 15530969 TI - Expression modulation of multiple cytokines in vivo by cyanobacteria blooms extract from Taihu Lake, China. AB - Cyanobacterial blooms that generate microcystins (MCs) are being increasingly recognized as a potent health hazard in aquatic ecosystems. However, immunomodulation induced by cyanotoxins has not been well documented. This paper reports the in vivo data on the immune disorder caused by crude microcystin (MC) extract of cyanobacteria blooms collected from Taihu Lake, China, with respect to cytokine mRNA levels. Using reverse-transcriptional polymerase chain reaction (RT PCR), the expression of multiple cytokines, including proinflammatory (IL-1beta, TNF-alpha, and IL-6) and Th1/Th2-related cytokines (IL-2, IL-4 and IL-10), was evaluated following the cyanobacteria blooms extract containing MCs (CBE) exposure at four doses of 23, 38, 77, 115 mg lyophilized algae cells/kg body weight. The results showed that the mRNA levels of TNF-alpha, IL-1beta, IL-2 and IL-4 decreased significantly following injection of all doses as compared to the control (LPS or ConA only), while the IL-6 level was unaffected. Contrast to this decrease, the level of IL-10 mRNA was, however, transiently up regulated following injection of the lowest dose of CBE. The distinct patterns of expression of these cytokines suggested a modulation of cytokine network, the essential component of the host immune system. We further developed a mathematical model to simulate the interaction of T helper cell subsets and related cytokines, which proved to be a good approach to study the kinetics of the interaction of cells and cytokines in microcystin immunosuppression. PMID- 15530970 TI - Determination of microcystin variants and related peptides present in a water bloom of Planktothrix (Oscillatoria) rubescens in a Spanish drinking water reservoir by LC/ESI-MS. AB - A water bloom of Planktothrix (Oscillatoria) rubescens was observed in a drinking water reservoir in central Spain in 2003. Microcystins where analysed by LC/ESI MS in 21 samples collected from this reservoir in five different days between March and May. A demethylated variant of microcystin-RR was identified as the major microcystin in most samples. Trace levels of microcystin-LR, -RR and -YR were detected in some samples. Four less common microcystins, with [M+H]+ ions at m/z 960, 981, 1045 and 1053, were also found. Total extracellular microcystin concentration varied from 0.010 to 19.126 microg l(-1). Furthermore, anabaenopeptins B and F as well as Oscillamide Y were also identified in these samples. PMID- 15530971 TI - Generation of active fragments from human zymogens in the bradykinin-generating cascade by extracellular proteases from Vibrio vulnificus and V. parahaemolyticus. AB - Vibrio vulnificus is an opportunistic human pathogen causing septicemia, and the infection is characterized by formation of the edematous skin lesions on limbs. This pathogenic species secretes a thermolysin-like metalloprotease as a virulence determinant. The metalloprotease was confirmed to activate human factor XII-plasma kallikrein-kinin cascade that results in liberation of bradykinin, a chemical mediator enhancing the vascular permeability, from high-molecular weight kininogen. Namely, the metalloprotease showed to generate active fragments by cleavage of Arg-Ile, Arg-Val or Gly-Leu peptide bond in human zymogens (plasma prekallikrein and factor XII). In spite of induction of the sufficient vascular permeability-enhancing and edema-forming reaction in the guinea pig model, a serine protease from V. parahaemolyticus, a human pathogen causing primarily watery diarrhea, showed far less ability to activate and to cleave the human zymogens. These results in part may explain why only V. vulnificus often causes serious edematous skin damages in humans. PMID- 15530972 TI - Two new actions of sea nettle (Chrysaora quinquecirrha) nematocyst venom: studies on the mechanism of actions on complement activation and on the central nervous system. AB - Chrysaora quinquecirrha (sea nettle) nematocyst venom is lethal to rainbow killifish (Adina xenica) when injected intraperitoneally or topically applied to the exposed brain or denuded epithelium. The lethal activity is thermostable requiring 100 degrees C heat for inactivation. This paper reports here for the first time that the venom also activates the complement system with the subsequent formation of the C5b-9 terminal complement complex. The events are associated with both a strong chemoattractant release and the tissue damage. These are also, at least in part, responsible for the pathogenesis of some clinical signs and symptoms associated to the jellyfish stings. PMID- 15530973 TI - Solid phase adsorption toxin tracking (SPATT): a new monitoring tool that simulates the biotoxin contamination of filter feeding bivalves. AB - A simple and sensitive in situ method for monitoring the occurrence of toxic algal blooms and shellfish contamination events has been developed. The technique involves the passive adsorption of biotoxins onto porous synthetic resin filled sachets (SPATT bags) and their subsequent extraction and analysis. The success of the method is founded on the observation that during algal blooms significant amounts of toxin, including the low polarity lipophilic compounds such as the pectenotoxins and the okadaic acid complex toxins, are dissolved in the seawater. The results of field trials during Dinophysis acuminata and Protoceratium reticulatum blooms are presented. These data prove the concept and demonstrate that the technique provides a means of forecasting shellfish contamination events and predicting the net accumulation of polyether toxins by mussels. As an early warning method it has many advantages over current monitoring techniques such as shellfish-flesh testing and phytoplankton monitoring. In contrast to the circumstantial evidence provided by genetic probe technologies and conventional phytoplankton monitoring methods, it directly targets the toxic compounds of interest. The extracts that are obtained for analysis lack many of the extraneous lipophilic materials in crude shellfish extracts so that many of the matrix problems associated with chemical and biological analysis of these extracts are eliminated. Analyses can confidently target parent compounds only, because analytical and toxicological uncertainties associated with the multiplicity of toxin analogues produced by in vivo biotransformation in shellfish tissues are reduced. Time integrated sampling provides a good simulation of biotoxin accumulation in filter feeders and the high sensitivity provides lengthy early warning and conservative estimates of contamination potential. The technique may reduce monitoring costs and provide improved spatial and temporal sampling opportunities. When coupled with appropriate analytical techniques (e.g. LC-MS/MS multi-toxin screens, ELISA assays, receptor binding assays), the technique has the potential to offer a universal early warning method for marine and freshwater micro-algae toxins. PMID- 15530974 TI - Studies of polyether toxins in the marine phytoplankton, Dinophysis acuta, in Ireland using multiple tandem mass spectrometry. AB - Diarretic shellfish poisoning (DSP) is a toxic syndrome associated with the consumption of bivalve molluscs. The DSP toxins are polyether compounds, which include okadaic acid (OA), dinophysistoxins (DTXs), pectenotoxins (PTXs) and pectenotoxin seco acids (PTX2SAs). These toxins originate in marine dinoflagellates, including Dinophysis spp. Phytoplankton samples were collected from the southwest coast of Ireland and D. acuta was the predominant species. Monocultures of D. acuta cells were prepared by hand picking from microscope slides in order to confirm their toxin profiles. There was a remarkable consistency in the toxin profiles in all of the phytoplankton samples collected during the summer months, irrespective of location, depth or mesh size. Analysis using liquid chromatography-multiple tandem mass spectrometry (LC-MS/MS) revealed that DTX2 and OA were the predominant toxins at a consistent ratio. The average toxin composition was: DTX2 (53+/-5%), OA (26.5+/-2.3%) and total pectenotoxins (20.8+/-4.7%). Toxin profiles in D. acuta from Europe were distinctly different from those found in New Zealand, where PTX2 was the predominant toxin and DTX2 was absent. PMID- 15530975 TI - Effect of microcystin-LR on protein phosphatase activity and glycogen content in isolated hepatocytes of fed and fasted juvenile goldfish Carassius auratus L. AB - Toxic effects of microcystin-LR were studied in hepatocytes isolated from fed and fasted juvenile goldfish Carassius auratus (30 g body weight). The hepatocytes were incubated with 10 microgMC-LR l(-1) during 4 h. MC-LR induces no effect in terms of cell number and viability. The toxin accumulation pattern was different in fed and fasted treatments. MC-LR accumulated more rapidly in 'fasted' cells where the highest concentration was observed by 1 h of exposure. It was delayed to 4 h in the 'fed' cells. MC-LR accumulation induced a severe decrease in hepatic protein phosphatase activity in both treatments. It was almost totally inhibited in both treatments during the first hour of exposure. The glycogen content was significantly reduced after 2 h of exposure in the fasting treatments, but not in the feeding one. PMID- 15530976 TI - Secretion and regeneration of tetrodotoxin in the rough-skin newt (Taricha granulosa). AB - Rough-skin newts (Taricha granulosa) released tetrodotoxin (TTX) in their skin secretions in response to mild electric stimulation. This release resulted in a large (21% to almost 90% of the pre-stimulation levels) reduction in the amount of TTX present in the dorsal skin of individual newts. Over the next 9 months newts significantly regenerated the levels of TTX in their skin. These data, in combination with previously published results, are consistent with the hypothesis that these newts produce their own TTX. PMID- 15530978 TI - Is there a risk of human poisoning by azaspiracids from shellfish harvested at the Portuguese coast? AB - Azaspiracid poisoning (AZP), the most recently discovered human gastrointestinal illness resulting from consumption of contaminated shellfish, so far has been found in coastal areas of northern Europe. This is the first report of a survey carried out for contamination of shellfish harvested in costal areas of Portugal for the presence of azaspiracids. The study design covered the commercial species usually more contaminated by toxins from dinoflagellates (blue mussel, common cockle, donax clam) in coastal areas representative of the NW, SW and south coasts. A method based on liquid chromatography-mass spectrometry was setup for the first time for this purpose. No azaspiracids were found on 300 samples tested between 2002 and 2003. On at least three samples a peak with a retention time matching that of AZA2 was found, never surpassing one tenth of the current EU limit. Unambiguous identification of any known AZA did not occur yet. The risk for human outbreaks of AZP seems to be very low, comparatively with amnesic shellfish poisoning (ASP), where levels close to the allowance level are found sparsely, or to diarrhetic shellfish poisoning (DSP) and paralytic shellfish poisoning (PSP), where high levels and registered human outbreaks have been found in recent years. PMID- 15530977 TI - Endogenous regulation of the functional duality of pahutoxin, a marine trunkfish surfactant. AB - Pahutoxin (PHN) is a long chain detergent-like quaternary ammonium cationic substance derived from defensive skin secretions of trunkfish. A recent study has revealed that PHN's ichthyotoxicity and its phospholipid membrane disruption effect are produced by two separate mechanisms, which presumably represent two separate physicochemical domains in the PHN molecule [Kalmanzon, E., Rahamim, Y., Barenholz, Y., Carmeli, S., Zlotkin, E., 2003. Receptor-mediated toxicity of pahutoxin, a marine trunkfish surfactant. Toxicon 42, 63-71]. Here we report on the occurrence of a natural endogenous mechanism, which regulates the above PHN's functional duality. The regulation is performed by the aid of two separates constituents of the trunkfish secretion, which either selectively amplify PHN's ichthyotoxicity (factor I) or suppress its phospholipids permeabilization effect (factor II). The ecological significance of such endogenous regulation is discussed. PMID- 15530979 TI - Molecular cloning of toxins expressed by the venom gland of Lasiodora sp. AB - The present work describes the identification of toxins expressed by the venom gland of the spider Lasiodora sp. The toxins LTx1, LTx2 and LTx3 were identified by the screening of a cDNA library. These toxins showed significant similarity at the amino acid level with spider toxins from Lasiodora parahybana, Eurypelma californicum, Brachypelma smithii, Selenocosmia huwena. PMID- 15530980 TI - A novel approach to quantitative ultrasonic naked gene delivery and its non invasive assessment. AB - The purpose of this study was to investigate practical, safe, easy-to-use, non cytotoxic, and reliable parameters to apply to an ultrasound (US) naked gene therapy system. The ultrasound pressure at the point of cell exposure was measured using a calibrated hydrophone and the intensity calculated. An acoustic power meter calibrated using a hydrophone was used to measure the power of the transducer. Four cell types were exposed to US with different exposure times and intensities. Fluorescent microscopy, spectrophotometry, scanning electron microscope, laser scanning confocal microscopy, flow cytometry and histogram analysis were used to evaluate the results of the study. The plasmid of green fluorescent protein (GFP) served as the reporter gene. The energy accumulation E in US gene delivery for 90% cell survival was defined as the optimal parameters (E=3.56+/-0.06), and at 80% cell survival was defined as the damage threshold (E=59.67+/-3.54). US safely delivered GFP into S180 cells (35.1 kHz) at these optimal parameters without obvious damage or cytotoxity in vitro. Exposed cell function was proved normal in vivo. The transfection rate was 35.83+/-2.53% (n=6) in viable cells, corresponding to 90.17+/-1.47% (n=6) cell viability. The intensity of GFP expression showed a higher fluorescent peak in the group of adeno-associated virus GFP vector (AVV-GFP) than in the control group (P<0.001). The effect of US gene delivery and cell viability correlated as a fifth order polynomial with US intensity and exposure time. With optimal parameters, US can safely deliver naked a gene into a cell without damage to cell function. Both optimal uptake and expression of gene depend on the energy E at 90% cell survival. E can be applied as a control factor for bioeffects when combined with other parameters. Stable caviation results in optimal parameters for gene delivery and the transient caviation may cause cell damage, which will bring about a sharp rise of permeabilization. The results may be applied to the development of a novel clinical gene therapeutic system. PMID- 15530981 TI - Multi-frequency harmonic arrays: initial experience with a novel transducer concept for nonlinear contrast imaging. AB - Nonlinear contrast imaging modes such as second harmonic imaging (HI) and subharmonic imaging (SHI) are increasingly important for clinical applications. However, the performance of currently available transducers for HI and SHI is significantly constrained by their limited bandwidth. To bypass this constraint, a novel transducer concept termed multi-frequency harmonic transducer arrays (MFHA's) has been designed and a preliminary evaluation has been conducted. The MFHA may ultimately be used for broadband contrast enhanced HI and SHI with high dynamic range and consists of three multi-element piezo-composite sub-arrays (A C) constructed so the center frequencies are 4f(A) = 2f(B) = f(C) (specifically 2.5/5.0/10.0 MHz and 1.75/3.5/7.0 MHz). In principle this enables SHI by transmitting on sub-array C receiving on B and, similarly, from B to A as well as HI by transmitting on A receiving on B and, likewise, from B to C. Initially transmit and receive pressure levels of the arrays were measured with the elements of each sub-array wired in parallel. Following contrast administration, preliminary in vitro HI and SHI signal-to-noise ratios of up to 40 dB were obtained. In conclusion, initial design and in vitro characterization of two MFHA's have been performed. They have an overall broad frequency bandwidth of at least two octaves. Due to the special design of the array assembly, the SNR for HI and SHI was comparable to that of regular B-mode and better than commercially available HI systems. However, further research on multi-element MFHA's is required before their potential for in vivo nonlinear contrast imaging can be assessed. PMID- 15530982 TI - Combining Brillouin spectroscopy and laser-SAW technique for elastic property characterization of thick DLC films. AB - Surface Brillouin spectroscopy (SBS) has been widely used for elastic property characterization of thin films. For films thicker than 500 nm, however, the wavelength of surface acoustic wave in the frequency range available for SBS is smaller than film thickness, and the SBS measures only the Rayleigh wave of the film. The laser-SAW technique, on the other hand, measures only the low-frequency portion of the surface acoustic wave dispersion and can estimate only one elastic modulus of the film (typically Young's modulus). In this work, we have combined the two methods to determine both Young's modulus and Poisson's ratio of a diamond-like carbon (DLC) film. It was found that reasonable estimates can be obtained for the longitudinal wave velocity, shear wave velocity, and Young's modulus of the film. The Poisson's ratio, however, still has a relatively large measurement error. PMID- 15530983 TI - Heat generation by impulse ultrasound. AB - An original method allowing to get a system of nonlinear evolution equations for the interacting modes applies to a problem of the heat generation by non-periodic ultrasound, including impulse one. The basic idea and final equations for the thermoviscous plane flow are presented. The limit of periodic source is traced. The numerical calculations were based on the pulse solution of the Burgers equation as an ultrasound source. Some illustrations on temporal behavior of the medium expansion caused by the pulse ultrasound are presented. PMID- 15530984 TI - Ultrasonic waves in diluted and densified suspensions. AB - A theory of propagation of stress waves in diluted and densified suspensions is developed to make the theoretical basis for analysis of ultrasonic waves through these media. The formulae for the phase velocity and the attenuation coefficient are determined as the function of wave frequency and the suspension structure parameter, which is the volume or mass fraction of the solid phase. These formulae can be use, after suitable calibration, for determination of the solid volume fraction in diluted suspensions, and the solid mass fraction or the water content in densified suspensions, that is, parameters that characterize the structure of a suspension. These structure parameters can be determined by measuring the transition time of ultrasonic wave through a given distance of suspension. The phase velocity dispersion curves and the attenuation coefficients determined theoretically and experimentally are plotted as a function of the volume fraction of the solid phase for dilute suspension, or the solid mass fraction for densified suspension. PMID- 15530985 TI - A numerical investigation of the resonance of gas-filled microbubbles: resonance dependence on acoustic pressure amplitude. AB - The general Keller-Herring equation for free gas bubbles is augmented by specific terms to describe the elasticity, viscosity and thickness of the encapsulating shell in ultrasound contrast agent microbubbles. A numerical investigation that analyses the acoustic backscatter from bubbles is employed to identify resonance frequencies that can be compared, for increasing driving pressure amplitude, with linear approximations obtained via analytical considerations. Calculations for bubbles of the size employed in diagnostic ultrasound, between 2 and 6 mum diameter, that are immersed in water and blood and exposed to monochromatic insonation, causing the bubbles to undergo stable cavitation, reveal that the resonance frequency diverges from the linear approximation as the pressure amplitude is increased. The shift in resonance, to lower frequency values, is found to be more pronounced for larger bubbles with the calculated value differing by up to 40% from the linear approximation. The results of this simulation might be potentially useful in preparation of formulations of ultrasound contrast agents with the specifically desired features, such as for instance resonance frequency. PMID- 15530986 TI - Non-destructive evaluation of elastic material properties in anisotropic circular cylindrical structures. AB - In this paper, non-axisymmetric guided wave propagation in circular cylindrical, anisotropic structures is studied in a frequency range up to 1 MHz. The investigations are carried out with carbon fibre reinforced tubes. The aim is the experimental determination of their effective linear elastic material properties in a non-destructive way. Therefore, an analytical model of the dispersion equation is fitted to the experimentally detected dispersion curves by systematically adjusting the desired material properties. A total least square scheme accompanied by an outlier detection criterion is used for this optimization task. Since the raw data of the measured dispersion curves contain a lot of noise, these outliers have to be detected and excluded, to achieve accurate results. Good agreement is found between the measured curves and the analytically calculated curves based on the estimated parameters. This fact indicates a high accuracy of the determined material properties. PMID- 15530987 TI - Chemical cross-linking and protein-protein interactions-a review with illustrative protocols. AB - The general term "protein-protein" interactions refers to the effects of proteins upon each other. The interactions can arise from co-existence in organized structural arrangements or in transient encounters. The latter are difficult to detect and define. Introduction of specific, stable chemical linkages can establish permanent relationships between what would normally be transiently associated species. The review covers the types and purposes of various linkers, including the comparative advantages of various approaches. The emphasis is on practical applications and thus includes methodology in the form of practical protocols for introducing the linkages and interpreting the outcomes. PMID- 15530988 TI - Synthesis of phosphonic analogues of carnitine and gamma-amino-beta hydroxybutyric acid. AB - The involvement of carnitine and gamma-amino-beta-hydroxybutyric acid in the biology of mammalian cells, the physiology of the human body, and some important aspects of medicinal treatment has induced many research groups to develop their pharmacologically potent analogues. Among them are the very important phosphonic analogues: phosphocarnitine and gamma-amino-beta-hydroxypropylphosphonic acid. This mini-review describes the various methodologies used for the synthesis of these compounds. PMID- 15530989 TI - 1-Deoxy-D-xylulose 5-phosphate reductoisomerase: an overview. AB - The methylerythritol phosphate pathway to isoprenoids, an alternate biosynthetic route present in many bacteria, algae, plants, and the malarial parasite Plasmodium falciparum, has become an attractive target for the development of new antimalarial and antibacterial compounds. The second enzyme in this pathway, 1 deoxy-D-xylulose 5-phosphate reductoisomerase (DXR; EC 1.1.1.267), has been shown to be the molecular target for fosmidomycin, a promising antimalarial drug. This enzyme converts 1-deoxy-D-xylulose 5-phosphate (DXP) into the branched compound 2 C-methyl-D-erythritol 4-phosphate (MEP). The transformation of DXP into MEP requires an isomerization, followed by a NADPH-dependent reduction. The discovery of DXR, its subsequent characterization, and the identification of inhibitors will be presented. PMID- 15530990 TI - Synthesis and biological evaluation of 4-imidazolylflavans as nonsteroidal aromatase inhibitors. AB - A series of 4-imidazolylflavans having a variety of substituents on the 2-phenyl ring was synthesized and investigated for their inhibitory effect against aromatase. Structure-activity relationships of these compounds were determined. An additional chlorine atom or a cyano group at the 4' position did not enhance aromatase inhibition as well as a 3'-hydroxyl group. The influence of an additional 4'-hydroxyl group depends on the substitution pattern of A ring. Among these molecules, 4'-hydroxy-4-imidazolyl-7-methoxyflavan is only 2.2-fold less active than the letrozole (as assessed by IC50 values). Letrozole is used as the first-line therapy for metastatic breast cancer. PMID- 15530991 TI - The role of conformational flexibility of enzymes in the discrimination between amino acid and ester substrates for the subtilisin-catalyzed reaction in organic solvents. AB - To investigate how the conformational flexibility of subtilisin affects its ability to discriminate between enantiomeric amino acid and ester substrates for the subtilisin-catalyzed reaction in an organic solvent, the flexibility around the active site and the surface of subtilisin was estimated from the mobility of a spin label bound to subtilisin by ESR spectroscopy. Many studies on enzyme flexibility focus on the active site. Both the surface and active site flexibility play an important role in the enantioselectivity enhancement of the enzyme-catalyzed reaction. It was found, however, that the different behavior observed for the enantioselectivity between the amino acid and ester substrates could be correlated with the flexibility around the surface rather than the flexibility at the active site of subtilisin. In other words, for the ester substrates, the greater flexibility around the surface of subtilisin induced by a conformational change resulting from the presence of an additive such as DMSO is essential for the enantioselectivity enhancement. This model is also supported by the Michaelis-Menten kinetic parameters for each enantiomeric substrate. Our findings provide insight into the enantioselectivity enhancement for the resolution of enantiomers for enzyme-catalyzed reactions in organic solvents. PMID- 15530992 TI - The effect of glycation on the chemical and enzymatic stability of the endogenous opioid peptide, leucine-enkephalin, and related fragments. AB - Nonenzymatic glycation is a posttranslational modification of peptides and proteins by sugars, which, after a cascade of reactions, leads to the formation of a complex family of irreversibly changed adducts implicated in the pathogenesis of human diseases. The stability of the Amadori compounds, the last reversible intermediates, determines the further course of the reaction. To provide information concerning the fate of glycated opioid peptides introduced into human circulation, the enzymatic (80% human serum) and chemical (phosphate buffer) stability of three Amadori compounds related to the endogenous opioid pentapeptide, leucine-enkephalin (Tyr-Gly-Gly-Phe-Leu), and to its N-terminal fragments: N-(1-deoxy-D-fructos-1-yl)-l-tyrosyl-glycyl-glycyl-L-phenylalanyl-L leucine, N-(1-deoxy-D-fructos-1-yl)-L-tyrosyl-glycyl-glycine, and N-(1-deoxy-D fructos-1-yl)-L-tyrosine were investigated. The results obtained in human serum indicate that N-terminal glycation of leucine-enkephalin significantly enhances proteolytic stability. While leucine-enkephalin itself was rapidly degraded (t1/2 = 14.8 min), the glycated-derivative was slowly converted (t1/2 = 14 h) to the corresponding Amadori /compound of Tyr-Gly-Gly and Phe-Leu. In phosphate buffer, the rate of hydrolysis of the Amadori compounds depends on the structure and length of the peptide moiety as well as on the concentration of the phosphate buffer. The hydrolysis patterns for the Amadori compounds in phosphate buffer and in human serum were not the same and appear to be specific for each substrate. PMID- 15530993 TI - Kinetic analysis of inhibition of cAMP-dependent protein kinase catalytic subunit by the peptide-nucleoside conjugate AdcAhxArg6. AB - Kinetic analysis of the inhibition of the phosphorylation of Kemptide, (LRRASLG), catalyzed by the catalytic subunit of cAMP-dependent protein kinase, by a peptide nucleoside conjugate inhibitor AdcAhxArg6 was carried out over a wide range of ATP and peptide concentrations. A simple procedure was proposed for characterization of the interaction of this inhibitor with the free enzyme, and with the enzyme-ATP and enzyme-peptide complexes. The second-order rate constants, calculated from the steady-state reaction kinetics, were used for this analysis to avoid the complications related to the complex catalytic mechanism of the protein kinase catalyzed reaction. PMID- 15530994 TI - Nonnucleoside HIV-1 reverse transcriptase inhibitors; part 3. Synthesis and antiviral activity of 5-alkyl-2-[(aryl and alkyloxyl-carbonylmethyl)thio]-6-(1 naphthylmethyl) pyrimidin-4(3H)-ones. AB - A series of 6-naphthylmethyl substituted S-alkylated dihydroalkoxybenzyloxopyrimidine (S-DABO) analogues with a beta-carbonyl group on the C-2 side chain were synthesized. All of the new compounds were evaluated for their anti-HIV activities in MT-4 cells. The most active compound, 5-isopropyl-2 [(4'-methoxyphenylcarbonyl-methyl)thio]-6-(1-naphthylmethyl)pyrimidin-4(3H)-one showed activity against HIV-1 and against the double mutated strain of HIV(Y181C and K103N) in the micromolar range. Furthermore, some of the compounds are active against both HIV-1 and HIV-2 in cell culture. In view of the fact that the loss of antiviral activity of these compounds when tested against S0561945 was much less pronounced than the loss of activity of typical NNRTIs, it is concluded that some of the compounds might interfere with another target or act on reverse transcriptase in a different way than the typical NNRTIs. PMID- 15530995 TI - The role of erythromycin C-12 hydroxylase, EryK, as a substitute for PikC hydroxylase in pikromycin biosynthesis. AB - The substrate flexibility of the erythromycin C-12 hydroxylase from Saccharopolyspora erythraea, EryK, was investigated to test its potential for the generation of novel polyketide structures. We have shown that EryK can accept the substrates of PikC from Streptomyces venezuelae which is responsible for the hydroxylation of YC-17 and narbomycin. In a S. venezuelae pikC deletion mutant, EryK could catalyze the hydroxylation of YC-17 and narbomycin to generate methymycin/neomethymycin and pikromycin, respectively. Molecular modeling of the enzyme-substrate complex suggested the possible interaction of EryK with alternative substrates. The results indicate that EryK is flexible toward some alternative polyketides and can be useful for structural diversification of macrolides by post-polyketide synthase hydroxylation. PMID- 15530996 TI - Spectroscopic studies of methylglyoxal in water and dimethylsulfoxide. AB - Methylglyoxal is a highly reactive dicarbonyl compound, which reacts in vivo with biological macromolecules and thereby affects their structure and function. These changes are associated with complications during aging, diabetes and Alzheimer's disease as well as with growth inhibition in different tumors. Many enzymes are involved in the metabolism of methylglyoxal, but its true physiological role in metabolism and chemical properties are still obscure. In this study it was shown that methylglyoxal, during the freeze-drying of aqueous solutions, polymerizes into small polymeric structures which are stable in organic media such as dimethylsulfoxide. When re-exposed to water, the polymers are immediately transformed into the monomeric mono- and dihydrate forms of methylglyoxal. By NMR and UV spectroscopy, it was shown that solvent, temperature, and the amount of available water strongly influence the equilibrium of the different forms of methylglyoxal and thereby change its reactivity. 1H and 13C NMR spectroscopy were used to determine the structures of the different monomeric and oligomeric structures of methylglyoxal. PMID- 15530997 TI - Regulating the fluorescence intensity of an anthracene boronic acid system: a B-N bond or a hydrolysis mechanism? AB - An anthracene-based fluorescent boronic acid system developed by the Shinkai group has been widely used for the preparation of fluorescent sensors for carbohydrates. Such application is based on the significant fluorescence intensity increase of this system upon binding with a carbohydrate. The mechanism through which this fluorescence intensity change happens was originally proposed to go through a B-N bond formation mechanism, which masks the nitrogen lone pair electrons. However, our own fluorescence studies suggest a possible alternative mechanism for the fluorescence change upon the formation of a boronic acid (1a) complex with diols. In this new proposed mechanism, complex formation induces solvolysis, which results in the protonation of the amine nitrogen if the reactions are carried out in a protic solvent such as water. This protonation prevents the photoinduced electron transfer, resulting in reduced quenching of the anthracene fluorescence. Such a solvolysis mechanism is supported by evidence from various types of experiments and theoretical calculations. PMID- 15530998 TI - Synthesis and characterization of topologically linked single-stranded DNA rings. AB - We investigated the synthesis of linked-ring DNAs by two DNA-ligation-based methods. In the first method, two DNA oligonucleotides were associated through a duplex segment of more than a full helical turn. Circularization of the entwined oligonucleotides by T4 DNA ligase resulted in two linked-ring DNAs with a total yield of approximately 40%. In the second method, a DNA oligonucleotide was circularized over a circular DNA template, resulting in the formation of approximately 10% linked-ring product. The circular nature of linked-ring DNAs was verified with exonuclease digestion and the existence of topological linkages was demonstrated by analyzing the electrophoretic mobility pattern of DNA products obtained from the digestion of each linked-ring DNA using specific restriction endonucleases. A linked-ring DNA library in which one of the two rings contained random-sequence nucleotides was also constructed and tested for compatibility with in vitro selection. PMID- 15530999 TI - Commonly used muscle relaxant therapies for acute low back pain: a review of carisoprodol, cyclobenzaprine hydrochloride, and metaxalone. AB - BACKGROUND: Low back pain is a leading reason for primary care visits. Many treatment options are available, but some lack scientific support. OBJECTIVE: The aim of this review was to discuss the etiology of low back pain and the relative risks and benefits of muscle relaxants commonly prescribed for the management of back pain. METHODS: We searched Intercontinental Marketing Services data for January 2003 through January 2004 to determine the most commonly prescribed agents for the management of musculoskeletal pain. Carisoprodol, cyclobenzaprine hydrochloride, and metaxalone represented >45% of all such prescriptions. Cochrane Library, MEDLINE, and EMBASE databases were searched (time frame: 1960 through January 2004; search terms: back pain, carisoprodol, cyclobenzaprine, metaxalone, muscle relaxants, and pharmacotherapy) and reference lists of identified articles were hand-searched. RESULTS: Three trials of carisoprodol (N = 197) were located in the Cochrane Library database. Two double-blind, randomized, placebo-controlled trials evaluating the safety and efficacy of cyclobenzaprine hydrochloride (N = 1405) were identified in the literature. Three double-blind, placebo-controlled trials were identified for metaxalone (N = 428) in 2 reports. The types of adverse events seen with these agents involved the central nervous system, including drowsiness/sedation, fatigue, and dizziness. However, the efficacy of cyclobenzaprine hydrochloride was shown to be independent of its sedative effects, which were dose related. The potential for abuse with carisoprodol is of growing concern. CONCLUSIONS: Analgesic pain management for low back pain due to muscle spasm may be combined with a muscle relaxant. Cyclobenzaprine hydrochloride has the most recent and largest clinical trials demonstrating its benefit, but carisoprodol and metaxalone also appear to be effective. However, carisoprodol's usefulness is mitigated by its potential for abuse. PMID- 15531000 TI - Rosuvastatin in the management of hyperlipidemia. AB - BACKGROUND: Rosuvastatin is a new statin indicated to reduce elevated levels of total cholesterol, low-density lipoprotein cholesterol (LDL-C), and triglycerides and to increase levels of high-density lipoprotein cholesterol (HDL-C) in patients with primary hypercholesterolemia, mixed dyslipidemia, and homozygous familial hypercholesterolemia. OBJECTIVE: The purpose of this article was to review the pharmacology, clinical efficacy, and tolerability of rosuvastatin as monotherapy and combination therapy for patients with hyperlipidemia. METHODS: A literature review was conducted using the search term rosuvastatin to identify English-language peer-reviewed articles and abstracts in the MEDLINE and Current Contents databases (both 1966 to March 2004). Citations from available articles were reviewed for additional references, and selected information from the manufacturer was discussed. RESULTS: Rosuvastatin 10 to 40 mg/d reduced LDL-C by 43% to 63% (P < 0.05). Compared with other statins, rosuvastatin had the highest dose-to-dose potency in lowering LDL-C (reduction of 60% vs 50% with atorvastatin, 40% with simvastatin, 30% with pravastatin or lovastatin, and 20% with fluvastatin) and better efficacy in raising HDL-C (increase of approximately 10% vs approximately 5% with other statins; P < 0.05). Rosuvastatin enabled significantly more patients to achieve the National Cholesterol Education Program (NCEP) goals for LDL-C with lower doses (P < 0.05). Rosuvastatin was well tolerated. Incidences of myopathy and liver function test abnormalities were rare and comparable to those of other statins. Because it is not metabolized by the cytochrome P-450 enzymes, rosuvastatin had fewer clinically significant drug interactions compared with other statins. Studies to assess the effect of rosuvastatin on cardiovascular outcomes are ongoing. CONCLUSIONS: Clinical studies continue to demonstrate that achieving optimal levels of LDL-C is an important goal in reducing cardiovascular events. Recent evidence suggests the need for an even lower LDL-C goal than that being recommended by the NCEP Based on the studies included in this review, rosuvastatin may help patients achieve optimal goals early with lower dosages, thus reducing the need for dose titration or combination therapy. PMID- 15531002 TI - Association between congestive heart failure and hospitalization in patients with type 2 diabetes mellitus receiving treatment with insulin or pioglitazone: a retrospective data analysis. AB - BACKGROUND: Thiazolidinediones (TZDs) are widely used oral antihyperglycemic drugs that facilitate insulin action and increase insulin-stimulated glucose metabolism, thereby decreasing insulin resistance. However, concerns have been raised regarding the association between TZD use and a heightened risk for congestive heart failure (CHF). OBJECTIVE: This study used claims data to conduct a retrospective examination of the CHF risk in patients with type 2 diabetes mellitus and to compare the association with CHF in those receiving the TZD pioglitazone and those receiving insulin. METHODS: Patients with type 2 diabetes aged > or =18 years who had begun treatment with pioglitazone or insulin between January 1999 and December 2001 were identified using the PharMetrics Patient Centric database. The sample was restricted to patients for whom there were > or =12 months of data before the index date (date of the first prescription for pioglitazone or insulin) and > or =3 months of follow-up data. Patients receiving a diagnosis of CHF before the index date were excluded. The propensity score for receiving pioglitazone was estimated using logistic regression based on available observed patient characteristics. Patients receiving insulin were matched in a 1:1 ratio with patients receiving pioglitazone based on a difference of no more than +/-0.01 in the estimated propensity score for receiving pioglitazone therapy. CHF risk was examined using the Cox proportional-hazards model. RESULTS: After exclusion of ineligible patients, 1668 matched pairs of patients receiving pioglitazone or insulin were identified (50.9% men, 49.1% women; mean [SE] age, 51.2 [0.2] years). The 2-year crude incidence rate of CHF was significantly lower in the pioglitazone group compared with the insulin group (primary/secondary diagnosis of CHF, 2.0% vs 4.0%, respectively; P < 0.001; inpatient hospitalization for CHF, 0.7% vs 2.5%; P < 0.001). The hazard ratio for pioglitazone versus insulin was 0.501 (95% CI, 0.331-0.758; P = 0.001) for a primary or secondary diagnosis of CHF in any setting and 0.263 (95% CI, 0.135 0.511; P < 0.001) for any occurrence of an inpatient hospitalization for CHF. CONCLUSIONS: In this retrospective analysis of data from patients with type 2 diabetes, pioglitazone therapy was associated with significantly lower incidence rates of CHF and inpatient hospitalization compared with insulin therapy. PMID- 15531001 TI - Effects of rosuvastatin versus atorvastatin, simvastatin, and pravastatin on non high-density lipoprotein cholesterol, apolipoproteins, and lipid ratios in patients with hypercholesterolemia: additional results from the STELLAR trial. AB - BACKGROUND: Non-high-density lipoprotein cholesterol (HDL-C), apolipoprotein (apo) B, and lipid and apolipoprotein ratios that include both atherogenic and antiatherogenic lipid components have been found to be strong predictors of coronary heart disease risk. OBJECTIVE: The goal of this study was to examine prospectively the effects of rosuvastatin, atorvastatin, simvastatin, and pravastatin across dose ranges on non-HDL-C, apo B, apo A-I, and total cholesterol (TC):HDL-C, low-density lipoprotein cholesterol (LDL-C):HDL-C, non HDL-C:HDL-C, and apo B:apo A-I ratios in patients with hypercholesterolemia (LDL C > or =160 mg/dL and <250 mg/dL and triglycerides <400 mg/dL) in the Statin Therapies for Elevated Lipid Levels compared Across doses to Rosuvastatin (STELLAR) trial. METHODS: In this randomized, Multicenter, parallel-group, open label trial (4522IL/0065), patients > or =18 years of age received rosuvastatin 10, 20, 40, or 80 mg; atorvastatin 10, 20, 40, or 80 mg; simvastatin 10, 20, 40, or 80 mg; or pravastatin 10, 20, or 40 mg for 6 weeks. Pairwise comparisons were prospectively planned and performed between rosuvastatin 10, 20, and 40 mg and milligram-equivalent or higher doses of comparators. RESULTS: A total of 2268 patients were randomized to the rosuvastatin 10- to 40-mg, atorvastatin, simvastatin, and pravastatin groups. Fifty-one percent of patients were women, the mean (SD) age was 57 (12) years, and 19% had a documented history of atherosclerotic disease. Over 6 weeks, rosuvastatin significantly reduced non-HDL C, apo B, and all lipid and apolipoprotein ratios assessed, compared with milligram-equivalent doses of atorvastatin and milligram-equivalent or higher doses of simvastatin and pravastatin (all, P < 0.002). Rosuvastatin reduced non HDL-C by 42.0% to 50.9% compared with 34.4% to 48.1% with atorvastatin, 26.0% to 41.8% with simvastatin, and 18.6% to 27.4% with pravastatin. Rosuvastatin reduced apo B by 36.7% to 45.3% compared with 29.4% to 42.9% with atorvastatin, 22.2% to 34.7% with simvastatin, and 14.7% to 23.0% with pravastatin. The highest increase in apo A-I (8.8%) was observed in the rosuvastatin 20-mg group, and this increase was significantly greater than in the atorvastatin 40-mg and 80-mg groups (both, P < 0.002). CONCLUSION: Rosuvastatin 10 to 40 mg was more efficacious in improving the lipid profile of patients with hypercholesterolemia than milligram equivalent doses of atorvastatin and milligram-equivalent or higher doses of simvastatin and pravastatin. PMID- 15531003 TI - Cyclosporine in patients with steroid-resistant nephrotic syndrome: an open label, nonrandomized, retrospective study. AB - BACKGROUND: Steroid-resistant nephrotic syndrome (SRNS) with focal segmental glomerulosclerosis has emerged as a leading cause of end-stage renal failure (ESRF) in children and adults. In the past decade, immunosuppressive drugs such as cyclosporine (CsA) and cyclophosphamide have been introduced for the treatment of SRNS, but data on long-term clinical outcome (over years) are lacking. OBJECTIVE: The current study considered the clinical outcome of patients with SRNS who had been treated with CsA for >2 years. The primary objective was to evaluate renal function after years of treatment compared with nontreated or CsA resistant patients. A secondary objective was to identify renal effects related to the use of CsA, with a major emphasis on renal fibrosis. METHODS: In this open label, nonrandomized, retrospective study, the outcomes of patients of all ages with sporadic SRNS who had been followed up for >2 years (between 1970 and 2002) at 4 Italian clinical institutions were evaluated. Preliminary molecular screenings for genes encoding proteins of the slit-diaphragm (eg, podocin, nephrin, alpha-actinin) were performed to exclude inherited forms of sporadic SRNS. RESULTS: A total of 157 patients were studied; mutations were found in 18 patients (11%). Of the remaining 139 patients (84 men, 55 women; median [interquartile range (IQR)] age at onset of proteinuria, 12 [4-32] years), 84 (60%) were nontreated and 55 (40%) were treated with CsA. Of these 55 treated patients, 35 (64%) were found to be resistant (ie, persistence of proteinuria after 2 months) or intolerant (ie, malignant hypertension or worsening of renal function), and CsA was withdrawn. The median (IQR) durations of follow-up for CsA resistant and nontreated patients were 41 (23-92) and 48 (28-106) months, respectively. Twenty patients (36%) were responsive to CsA and were followed up for a median (IQR) of 81 (47-115) months. Progression Lo ESRF occurred in 10% of CsA-responsive patients versus 60% of CsA-resistant patients and 62% of nontreated patients (P = 0.002). No sign of renal fibrosis related to drug toxicity was observed in renal biopsies performed at 5-year intervals. CONCLUSIONS: This retrospective analysis of SRNS documented a persistent antiproteinuric effect of long-term CsA (>2 years) in the absence of renal fibrosis. Although sensitivity to CsA was associated with normal renal function, resistance or intolerance was associated with progression to ESRF These data suggest that CsA may have a role in the treatment of patients with SRNS. PMID- 15531004 TI - Efficacy and safety of delapril plus manidipine compared with enalapril plus hydrochlorothiazide in mild to moderate essential hypertension: results of a randomized trial. AB - BACKGROUND: The use of combination therapy is required to achieve blood pressure targets in 40% to 75% of patients with hypertension. There have been few studies comparing the efficacy and tolerability of the new fixed combination of the angiotensin-converting enzyme (ACE) inhibitor delapril 30 mg and the calcium channel antagonist manidipine 10 mg with those of a standard combination of another ACE inhibitor and a diuretic. OBJECTIVE: The aim of this study was to compare the antihypertensive efficacy and tolerability of delapril 30 mg given alone or with manidipine 10 mg with those of enalapril 20 mg given alone or with hydrochlorothiazide (HCTZ) 12.5 mg in patients with mild to moderate essential hypertension. METHODS: This was a multicenter, active-controlled, parallel-group trial. After an initial 2-week placebo run-in period, patients aged 18 to 75 years with diastolic blood pressure (DBP) > or =90 and < or =109 mm Hg were randomized in a 2:1 ratio to receive delapril or enalapril for 8 weeks. After the initial 8 weeks, nonresponders (DBP > or =85 mm Hg) received an additional 8 weeks of treatment with a fixed combination of delapril + manidipine or enalapril + HCTZ; patients whose DBP was normalized continued their initial monotherapy through the end of the study. The primary efficacy variable was the change in sitting DBP at the end of treatment. Secondary efficacy variables were the percentage of patients whose DBP was normalized (DBP Z:85 mm Hg) and the percentage of responders (> or =10-mm Hg reduction in DBP or DBP <85 mm Hg). RESULTS: One hundred sixty patients (84 men, 76 women) were randomized to receive delapril (n = 106) or enalapril (n = 54). After 16 weeks of treatment, the mean (SD) reduction in DBP was similar with the 2 treatments (delapril, -14 [8] mm Hg; enalapril, -15 [8] mm Hg). In the delapril and enalapril groups, DBP was normalized in a respective 55 (51.9%) and 29 (53.7%) patients, and 77 (72.6%) and 38 (70.4%) were responders; there was no significant difference between groups. Tolerability was also similar in both groups--10 (9.4%) patients in the delapril group and 5 (9.3%) in the enalapril group experienced adverse events that were judged related to treatment. CONCLUSIONS: The results of this study suggest that delapril alone or combined with manidipine is well tolerated and as effective as enalapril alone or combined with HCTZ in lowering blood pressure in patients with mild to moderate essential hypertension. PMID- 15531005 TI - Use of sibutramine in overweight adult hispanic patients with type 2 diabetes mellitus: a 12-month, randomized, double-blind, placebo-controlled clinical trial. AB - BACKGROUND: The management of type 2 diabetes mellitus is complicated by the presence of risk factors related to overweight and obesity, particularly visceral adiposity. However, weight loss and weight maintenance are difficult for patients with diabetes, and the benefits of dietary modifications are typically modest. Sibutramine is a serotonin- and norepinephrine-reuptake inhibitor that reduces food intake by inducing early satiety and attenuates the decrease in basal energy expenditure associated with weight loss. Previous trials of sibutramine in overweight and obese patients with type 2 diabetes have shown significant weight loss accompanied by better glycemic control. OBJECTIVE: The goal of this study was to assess the effect on body weight and glycemic control of sibutramine in combination with glibenclamide in obese Hispanic patients with type 2 diabetes. METHODS: This was a 12-month, randomized, double-blind, placebo-controlled clinical trial conducted at the Endocrinology Service, General Hospital of Mexico, Mexico City. Included were overweight or obese (body mass index [BMI] >27 kg/M2) patients with type 2 diabetes between the ages of 24 and 65 years who had been receiving glibenclamide monotherapy for at least 2 weeks and whose glucose concentrations were stable. Patients were randomized to receive sibutramine 10 mg or placebo once daily. The primary efficacy measures were change in body weight, waist circumference, and glycosylated hemoglobin (HbA1c). Anthropometrics and fasting glucose concentrations were measured monthly. HbA1c was determined at baseline and at 6 and 12 months. Laboratory parameters were measured at baseline and at the end of the study. RESULTS: Forty-four patients were randomized to receive sibutramine (28 women, 16 men; mean [SD] age, 47.6 [9.0] years), and 42 were randomized to receive placebo (31 women, 11 men; mean age, 45.8 [8.1] years). Twenty-four patients in the sibutramine group and 23 in the placebo group completed the trial. In the sibutramine group, body weight was reduced from a mean (SD) of 73.9 (10.3) kg at baseline to 69.8 (10.6) kg at month 12; BMI decreased from 29.9 (2.6) to 28.2 (2.9) kg/M2; waist circumference was reduced from 94.9 (8.4) to 90.8 (8.4) cm; the plasma fasting glucose concentration decreased from 140.4 (29.4) to 114.2 (32.0) mg/dL; and the HbA1c value was reduced from 8.9% (1.2) to 8.3% (1.2) (all, P < 0.001). In the placebo group, the corresponding changes were from 74.5 (10.3) kg at baseline to 73.1 (11.2) kg at month 12; from 30.1 (2.5) to 29.5 (2.9) kg/M2; from 94.4 (7.3) to 93.1 (8.3) cm (P < 0.05); from 140.7 (25.2) to 123.9 (38.3) mg/dL (P < 0.05); and from 9.0% (1.2) to 9.1% (1.3). In the sibutramine group, weight loss continued for up to 12 months. CONCLUSION: In this population of obese Hispanic patients with type 2 diabetes, sibutramine combined with glibenclamide therapy achieved weight loss for up to 12 months and was associated with better glycemic control than placebo. PMID- 15531006 TI - Ambulatory versus clinic blood pressure for the assessment of anti hypertensive efficacy in clinical trials: insights from the Val-Syst Study. AB - BACKGROUND: Several studies have found that measurement of blood pressure (BP) in the clinical setting may lead to overestimation of hypertension and may yield inaccurate assessments of the efficacy of antihypertensive treatment. OBJECTIVE: The aim of this study was to determine whether the use of clinic BP in the Valsartan and Amlodipine for the Treatment of Isolated Systolic Hypertension in the Elderly (Val-Syst) study accurately identified those elderly outpatients with systolic hypertension who had true 24-hour elevations in BP, as well as those who required dose increases in antihypertensive therapy during follow-up. METHODS: In Val-Syst, patients aged between 60 and 80 years with a clinic sitting systolic BP (SBP) of 160 to 220 mm Hg and a diastolic BP <90 mm Hg after a 2-week placebo washout period were randomized to receive valsartan 80 mg or amlodipine 5 mg once daily (level 1). In those with a trough SBP > or =140 mm Hg after 8 weeks of double-blind treatment, doses were titrated upward to valsartan 160 mg or amlodipine 10 mg once daily (level 2). If clinic SBP was > or =140 mm Hg after a further 8 weeks, hydrochlorothiazide 12.5 mg was added for an additional 8 weeks (level 3). Clinical decisions during the active-treatment period were based on clinic BP measurements. Thirteen of the 35 participating centers assessed ambulatory BP as well as clinic BP at baseline and the end of the treatment, making it possible to compare the results of the 2 modes of measurement. The Student test was used to compare drug-induced changes in clinic and ambulatory BP in individual patients. Differences between the decreases in clinic and ambulatory BP at the 3 treatment levels were tested using repeated-measures analysis of covariance (ANCOVA), with baseline as the covariate. RESULTS: One hundred sixty-four elderly patients (age range, 60-80 years; 85 men, 79 women) were included in the study (79 valsartan, 85 amlodipine), and valsartan and amlodipine were reported to have comparable effects on the level and rhythm of 24 hour BP In the present study, 22 of 164 patients had white-coat hypertension at baseline (clinic SBP > or =160 mm Hg and mean 24-hour SBP <130 mm Hg). For both treatments combined, the mean (SD) decreases in clinic SBP were inversely proportional to the treatment level (level 1 = -33.2 (7.9) mm Hg; level 2 = -31.6 (11.8) mm Hg; level 3 = -29.3 (11.6) mm Hg; P = 0.001, overall ANCOVA). In contrast, after adjusting for baseline values, the decreases in mean 24-hour SBP did not differ between treatment levels (level 1 = -10.8 [10.4] mm Hg; level 2 = 13.0 [11.2] mm Hg; level 3 = -16.4 [13.8] mm Hg). The decrease in clinic BP during therapy was similar in patients with white-coat hypertension and sustained hypertension (clinic SBP > or 160 mm Hg and mean 24-hour SBP > or =130 mm Hg), whereas 24-hour and 8- to 9-am SBP decreased significantly only in patients with sustained hypertension (P < 0.001). At the end of the study, mean 24-hour SBP continued to be uncontrolled (> or =130 mm Hg) in 16 of 53 patients (30.2%) at treatment level 1, 27 of 62 (43.5%) at level 2, and 19 of 49 (38.8%) at level 3 (P = NS). CONCLUSION: Based on the findings in this population of elderly patients with systolic hypertension, the management of hypertension may vary depending on whether decisions concerning the selection of patients for clinical trials and treatment adjustments during follow-up are made using clinic or ambulatory BP measurement. PMID- 15531007 TI - Incidence and duration of antidepressant-induced nausea: duloxetine compared with paroxetine and fluoxetine. AB - OBJECTIVE: This analysis assessed the incidence, severity, onset, and duration of nausea among patients with major depressive disorder (MDD) treated with the new antidepressant duloxetine. METHODS: Data were pooled from 8 double-blind, randomized, placebo- and active comparator-controlled trials employing patients with MDD that were submitted to the US Food and Drug Administration to support duloxetine's new drug application for treatment of MDD. RESULTS: The numbers of patients receiving each regimen were as follows: placebo, n = 777; duloxetine 40 mg/d, n = 177; duloxetine 60 mg/d, n = 251; duloxetine 80 mg/d, n = 363; duloxetine 120 mg/d, n = 348; paroxetine 20 mg/d, n = 359; and fluoxetine 20 mg/d, n = 70. In acute placebo-controlled trials of duloxetine 40 to 120 mg/d, treatment-emergent nausea was reported by more duloxetine-treated patients than those receiving placebo (19.9% [227/1139] vs 6.9% [154/777], respectively; P <0.001). Among duloxetine-treated patients, the median time to onset of nausea was 1 day, and the median duration of nausea was 7 days. The incidence of nausea was similar to placebo rates after 1 week. In paroxetine-controlled studies, the incidence of treatment-emergent nausea in patients receiving duloxetine did not differ significantly from paroxetine (14.4% vs 12.0%, respectively). In head-to head studies, the incidence of treatment-emergent nausea with duloxetine did not differ significantly from that with fluoxetine (17.1% vs 15.7%, respectively). Most duloxetine-treated patients reported nausea to be mild (52.9%) or moderate (41.4%). Treatment discontinuation secondary to nausea occurred in more duloxetine-treated patients than those receiving placebo (1.4% [16/1139] vs 0.1% [1/777], respectively; P = 0.002). Following abrupt discontinuation after acute treatment, 5.9% of duloxetine-treated patients exhibited nausea compared with 0.3% of patients receiving placebo (P < 0.001). The incidence of treatment emergent nausea during 6-month continuation of duloxetine treatment (80 mg/d, 2.1%; 120 mg/d, 1.3%) was similar to placebo (1.6%). Following abrupt discontinuation after 8 months of treatment, nausea was reported by 1.6% of patients receiving duloxetine 120 mg/d compared with 0% for those receiving duloxetine 80 mg/d and 0% for placebo. CONCLUSIONS: Duloxetine induced mild to moderate nausea in a subset of patients with MDD during treatment initiation. Nausea resolved rapidly with continued treatment. The incidence of duloxetine induced nausea resembled that produced by paroxetine and fluoxetine. PMID- 15531008 TI - A new fenofibrate formulation: results of six single-dose, clinical studies of bioavailability under fed and fasting conditions. AB - BACKGROUND: The main drawback of the lipid-lowering agent fenofibrate is low bioavailability when taken orally on an empty stomach. Even the newest marketed formulations, micronized fenofibrate (MF) 200-mg capsules and microcoated fenofibrate (MCF) 160-mg tablets, require administration with meals to increase bioavailability. Insoluble Drug Delivery-Microparticle (IDD-P) fenofibrate 160-mg tablets are a new formulation developed to provide bioavailability independent of food. OBJECTIVE: The aims of this group of studies were to determine the effects of food on the bioavailability of IDD-P fenofibrate and MCF and to compare the bioavailabilities of IDD-P fenofibrate, MF, and MCF under fasting and fed conditions. METHODS: Six single-dose pharmacokinetic studies were conducted in healthy adult volunteers. The primary end points were the extrapolated area under the curve (AUC) from time 0 to infinity (AUC(0-infinity)), the experimental AUC from time 0 to time t, and the maximum plasma concentration of the active fenofibrate metabolite, fenofibric acid. The bioavailabilities of tested treatments were compared using the 90% CIs of ratios of least squares means (RLSMs) of logarithmically transformed AUC(0-infinity) values; 90% CIs within 80% to 125% were considered to indicate equivalent bioavailability (ie, extent of absorption). RESULTS: One hundred thirteen subjects (87 men, 26 women) received > or =1 study treatment and were evaluable for tolerability, and 99 of them (77 men, 22 women) were evaluable for bioavailability. The RLSMs of IDD-P fenofibrate extent of absorption (AUC(0-infinity)) under fed versus fasting conditions were 104.99%, 118.76%, and 113.56% (high-fat fed) and 111.06% and 110.64% (low-fat fed) in 5 separate comparisons. The corresponding 90% CIs were 95.38% to 115.58%, 113.38% to 124.40%, 107.54% to 119.93%, 106.06% to 116.32%, and 104.75% to 116.86%. All of these CIs were within the 80% to 125% range, suggesting equivalent bioavailability of IDD-P fenofibrate independent of food. The RLSM for MCF under high-fat fed versus fasting conditions was 151.19% (90% CI, 123.59% 184.94%). CONCLUSIONS: In healthy adult volunteers, IDD-P fenofibrate had an equivalent extent of absorption under fed or fasting conditions, suggesting that dosage regimens could include administration of the product without food. Administering drug independently of food may provide greater convenience and simplicity for patients and prescribers. PMID- 15531009 TI - Results of an open-label, prospective study of anticoagulant therapy for atrial fibrillation in an outpatient anticoagulation clinic. AB - OBJECTIVE: The goal of this study was to investigate the complications and control of warfarin treatment in patients with nonvalvular atrial fibrillation (NVAF) newly referred to an outpatient anticoagulation clinic. METHODS: This study included new patients with NVAF who were referred to an anticoagulation clinic for warfarin therapy over a recruitment period of 21 months. To reflect real-world clinical practice, patient selection for anticoagulation and patient management were left to the referring physicians, who were blinded to their patients' participation in the study. Patients were interviewed in person at the first clinic visit and then by telephone every 4 to 6 weeks. They were questioned about any bleeding or thromboembolic events. RESULTS: A total of 402 patients were included (100% of all new referrals over 21 months). The mean (SD) age was 72.3 (10.3) years, and 224 (56%) patients were men. The mean (SD) international normalized ratio (INR) was 2.4 (0.31). Patients were followed up for a mean (SD) of 19 (8.1) months (range, 1.0-31.0 months). They spent a mean (SD) 66% (18.3) of time in the target range of INR (ie, 2.0-3.0). Annual event rates were 1.7% (95% CI, 0.4%-3.0%) for major bleeding, 16.6% (95% CI, 13.0%-20.2%) for minor bleeding, 1.2% (95% CI, 0.1%-2.3%) for ischemic stroke, and 0.3% (95% CI, 0.2% 0.8%) for transient ischemic attacks. There were no cases of hemorrhagic stroke or fatal bleeding. Variability of INR and number of medications were identified as risk factors for bleeding (P = 0.03 and P = 0.001, respectively). There was no significant association between age and bleeding. CONCLUSIONS: Based on this analysis, the risks of long-term oral anticoagulation therapy in an outpatient anticoagulation clinic appear to reflect the results of clinical trials. Rates of ischemic stroke, major bleeding, and anticoagulation control were comparable. There was no age-related risk of complications. PMID- 15531011 TI - Efficacy comparison between preprandial and postprandial insulin aspart administration with dose adjustment for unpredictable meal size. AB - BACKGROUND: Patients with type 1 diabetes mellitus (DM) may encounter situations in which meal size and content is unpredictable. In theory, postprandial injection of rapid-acting insulin analogues could prove more effective in achieving glucose control at such times because this treatment strategy could allow adjustment of insulin dose for the actual size of the meal consumed rather than being based on an estimate of what will be consumed. OBJECTIVE: This study compared the postprandial glycemic control achieved with meal-related insulin aspart injected immediately before a meal with that injected immediately after a meal. METHODS: This randomized, crossover study was conducted at Sansum Diabetes Research Institute, Santa Barbara, California. Adult patients with type 1 DM were enrolled. At study visit 1, patients were randomly assigned to inject insulin aspart 0 to 5 minutes before the start of the meal or immediately after the meal. The timing of injection relative to the meal was reversed at study visit 2. The meal-related dose was calculated based on the anticipated caloric and carbohydrate intake (preprandial injection) or actual calories and carbohydrates ingested (postprandial injection). Postprandial blood glucose concentrations were evaluated as markers of efficacy of postprandial aspart administration. RESULTS: Twenty patients were enrolled in the study (mean [SE] duration of DM, 22.5 [3.2] years; mean [SE] body mass index, 26.2 [1.0] kg/M2; age range, 22-82 years); 19 completed it. Total glucose AUC during the meal test was 22% less when insulin aspart was injected immediately before the study meal (mean [SE], 23,014 [1832] mg/dL.min) than when injected immediately after the meal (mean [SE], 29,535 [2243] mg/dL.min) (P < 0.001), but baseline-adjusted AUC was similar. Maximum mean (SD) glucose concentrations in the postprandial period were <180 mg/dL, the current DM treatment goals specified by the American Diabetes Association (149.0 [9.9] mg/dL and 102.0 [9.2] mg/dL, following postprandial insulin aspart injection and preprandial injection, respectively; P < 0.001). There was variation in the number of calories consumed, but patients consumed a similar number of calories in the 2 treatment regimens. The frequency of postprandial hypoglycemia was comparable. Adjustment of postprandial insulin aspart dose for the actual meal size consumed maintained postprandial glucose concentrations within currently recommended treatment guidelines. CONCLUSIONS: Preprandial insulin aspart injection produced a better glucose profile and is preferred when conditions permit. However, both preprandial and postprandial insulin aspart administration achieved postprandial glucose concentrations within currently recommended treatment guidelines. PMID- 15531010 TI - Open-label, dose-ranging pilot study of 4 weeks of low-dose therapy with sodium phosphate tablets in chronically constipated adults. AB - BACKGROUND: The tablet formulation of sodium phosphate (NaP) is a prescription osmotic purgative that has been marketed since 2001. The use of NaP tablets in patients with constipation has not been studied previously. OBJECTIVE: This study assessed the tolerability and efficacy of 28 days of therapy with NaP tablets (1.5 g NaP/tablet) in patients with chronic constipation. METHODS: Adults with functional constipation or constipation-predominant irritable bowel syndrome and Z-3 spontaneous bowel movements (BMs) during the 7-day screening/baseline period were eligible for this open-label, dose-ranging study. Patients were randomized to receive starting doses of 4 NaP tablets (group A) or 8 NaP tablets (group B) each morning for 28 days. After a minimum of 48 hours, the NaP dose could be titrated upward (in the case of no BM or no relief of symptoms) or downward (in the case of a predefined excess laxative response) by 2 tablets/d to a minimum of 2 tablets/d or a maximum of 12 tablets/d. Patients kept a diary of their BMs and gastrointestinal symptoms. A serum chemistry panel was obtained weekly. The primary end points were the constipation response (based on the change from baseline in weekly number of BMs) and the global sense response (based on daily scores for the patient's overall sense of change in their bowel problems). RESULTS: At randomization, there were 18 patients in group A and 25 in group B. Of these, 40 patients (16 group A, 24 group B) had > or 7 days of diary information while taking study treatment and were evaluable for efficacy. The constipation response rate was 100% in group A and 95.8% in group B, and the respective global sense response rates were 68.8% and 79.2%. Four patients in group B withdrew due to adverse events, none of which were serious. Five patients had occasional hypokalemia that required no treatment. Changes from baseline in serum concentrations of calcium, inorganic phosphorus, and potassium were not clinically significant and did not require treatment. CONCLUSIONS: In this small study, NaP tablets taken daily were generally well tolerated (particularly in the low-dose group) and produced prompt relief of constipation--generally within the first week of treatment--that was sustained over the 28-day treatment period. A reasonable starting dose appears to be 2 to 4 tablets (3-6 g NaP) daily. PMID- 15531012 TI - Preference for insulin delivery systems among current insulin users and nonusers. AB - BACKGROUND: It is not known whether preferences for the vial and syringe (VS) or the insulin injection pen device (IIPD) differ between current insulin users and nonusers. Additional benefits in treatment might be realized if preference were considered when discussing insulin use with patients initiating or changing insulin treatment. OBJECTIVE: The objective of this study was to examine respondent preferences for the VS and the IIPD between current insulin users and nonusers. METHODS: US residents with type 1 or 2 diabetes mellitus who responded to a year-2001 mail survey completed a 19-item self-administered questionnaire designed to assess respondents' expectations of attributes related to the VS and IIPD. Items were analyzed on a 5-point Likert-type scale with higher scores indicating greater agreement that attributes met expectations. Composite scores for ease of use, activity interference, and social acceptability were used to further examine differences between insulin users and nonusers regarding their preferences for the VS or IIPD. Observed differences in preferences were evaluated statistically using the chi-square test, paired Student t test, and regression analysis. RESULTS: Questionnaires were received from 302 respondents, producing an adjusted response rate of 20.8%. Respondents ranged in age from 18 to 83 years (mean [SD], 52.4 [13.2] years), with 62% reporting annual income above 25,000 US dollars. Of the 242 usable responses, 99 respondents were insulin users and 143 were not. Statistical evaluation using analysis of variance revealed significant regression coefficients (P < or = 0.001) for both insulin users and nonusers for each of the 3 dimensions (ease of use, activity interference, and social acceptability with respect to preference). CONCLUSIONS: Based on this survey analysis, overall preference for the IIPD appeared to be higher compared with the VS among both insulin users and nonusers. Social acceptability was the strongest predictor of preference for the IIPD. For current insulin users, social acceptability and ease of use were significant predictors of preference for the VS. For insulin nonusers, these results suggested that patient discussions about VS should emphasize activity interference and ease of use. PMID- 15531014 TI - The political calculus of a Medicare prescription drug benefit. PMID- 15531013 TI - Dose-response relationship with increasing doses of dextromethorphan for children with cough. AB - BACKGROUND: The efficacy of dextromethorphan (DM) for treating acute cough is uncertain, and its use is not supported by the American Academy of Pediatrics. Nevertheless, DM is often administered to children as an antitussive. DM dosages are based on age rather than body weight, resulting in substantial variability in the relative amount of drug administered. OBJECTIVE: The aim of this work was to determine whether a dose-response relationship existed among a group of children administered a single nocturnal dose of DM for cough due to an upper respiratory tract infection. METHODS: As part of a larger double-blind, placebo-controlled trial of over-the-counter cough medications, children received DM. The administered doses (per manufacturer recommendations) were as follows: ages 2 to 5 years, 7.5 mg; ages 6 to 11 years, 15 mg; and ages 12 to 18 years, 30 mg. This resulted in a range of 0.35 to 0.94 mg/kg per dose. Subjective parental assessments of cough and sleep were obtained using a 7-point Likert-type scale that compared symptoms after medication with symptoms during the prior night (without medication). Three dose ranges were compared as a subset analysis of the group that received DM. RESULTS: Thirty-three patients (19 girls, 14 boys; median [interquartile range] age, 4.90 [2.90-6.80] years; age range, 2.10-16.50 years) received DM and completed the study. No significant differences were found for any of the outcome measures when comparing the effects of different doses of DM, but our observations suggested somewhat more symptomatic relief for patients receiving medium-dose DM (0.45 to <0.60 mg/kg per dose) or high-dose (HD) DM (0.60-0.94 mg/kg per dose) compared with low-dose DM (0.35 to <0.45 mg/kg per dose). Adverse events occurred most often in the HD group. CONCLUSIONS: Although no statistically significant differences were detectable for the outcomes studied, our observations suggest the potential for improved clinical symptom control with increasing doses of DM. Our findings may further suggest that a dose of 0.5 mg/kg should be considered in future assessments of the antitussive effect of DM in pediatric studies, to balance symptomatic relief with the avoidance of adverse events. PMID- 15531015 TI - Effects of a prior-authorization policy for celecoxib on medical service and prescription drug use in a managed care Medicaid population. AB - BACKGROUND: Prior authorization (PA) is a poorly studied but commonly employed policy used by health care payers to manage the rising costs of pharmacy benefits. OBJECTIVE: The aim of this study was to evaluate the intended and unintended effects of a PA policy for celecoxib on pharmacy and medical-service utilization in a Medicaid managed-care organization. METHODS: This was a retrospective, interrupted time-series analysis of 22 monthly health-related utilization rates from January 1, 1999, to October 31, 2000. All Medicaid claims for CareOregon (a managed-care organization) and a fee-for-service program were reviewed. A model was constructed to evaluate changes in utilization of therapeutically related drug classes (eg, conventional nonsteroidal anti inflammatory drugs [NSAIDs], gastrointestinal agents), office and emergency department encounters, and hospitalizations before and after the PA policy was implemented on November 16, 1999. A secondary analysis evaluated these changes among a sample of prior NSAID users. RESULTS: After the PA policy was implemented, use of celecoxib was immediately reduced from 1.07 to 0.53 days' supply per person-year (58.9%; 95% CI, 50.0%-67.9%). The monthly rate of increase was also reduced (P < 0.001). Utilization changes were not observed in other drug classes. Similar changes were observed in the secondary analysis. An 18% (95% CI, 2.2%-33.9%) nonsignificant increase in emergency-department visits was observed in the entire sample after the PA policy was implemented. However, a similar change was not observed in the secondary analysis of prior NSAID users. No other changes in medical service encounters were noted after the PA policy was activated. CONCLUSIONS: This observational study found that celecoxib use was substantially reduced after the implementation of a PA policy. No important changes in use of other drug classes were detected. The overall increase in emergency-department visits--although not observed among previous NSAID users- should be explored on the individual level. PMID- 15531016 TI - Priorities for triptan treatment attributes and the implications for selecting an oral triptan for acute migraine: a study of US primary care physicians (the TRIPSTAR Project). AB - BACKGROUND: Physicians treating patients with migraine can now choose from among 7 triptans, which differ on a range of attributes that may be important for treatment selection. OBJECTIVE: The aims of this study were to determine the relative importance of treatment attributes of the available triptans and assess their impact on deciding the most appropriate treatment for a particular patient with migraine. METHODS: As part of the TRIPSTAR project, US primary care physicians were surveyed to elicit their views on the relative importance of a prespecified set of treatment attributes for making treatment choices in clinical practice. The treatment attributes assessed were those for which data from controlled clinical trials were available for subsequent comparison. The resulting attribute-importance weights were then combined with data on the performance of individual triptans across these attributes in a multiattribute decision model to assist selection of an oral triptan. RESULTS: Efficacy attributes were considered more important than tolerability or consistency of effect in selecting an oral triptan. For triptan-naive but not triptan experienced patients, tolerability was considered significantly more important than consistency (30% [95% CI, 27%-34%] vs 21% [19%-24%]). Sustained pain-free status and freedom from cardiovascular (chest) adverse events were the most important efficacy and tolerability attributes for both patient categories. When the relative importance of the treatment attributes was combined in a multiattribute decision model with meta-analysis data from controlled trials, almotriptan, eletriptan, and rizatriptan were significantly closer to the hypothetical ideal triptan than the reference product, sumatriptan 100 mg. CONCLUSION: Multiattribute decision-making models (such as that used in the TRIPSTAR project) that determine and apply the relative importance of treatment attributes to drug selection have considerable potential value as a decision support tool in the treatment of acute migraine. PMID- 15531017 TI - Cost utility and budget impact of third-generation aromatase inhibitors for advanced breast cancer: a literature-based model analysis of costs in the Italian National Health Service. AB - BACKGROUND: Third-generation aromatase inhibitors are effective alternatives to tamoxifen in patients with advanced breast cancer. However, their acquisition costs might burden fixed-budget health care systems. OBJECTIVE: This study is a decision analysis of the clinical and economic consequences of alternative first line hormone therapies for postmenopausal women with estrogen receptor-positive metastatic breast cancer in a real-life Italian health care setting. METHODS: A Markov model was developed to describe disease evolution according to data from previously published, randomized clinical trials. The costs incurred by a local community hospital in the Italian National Health Service were considered (year 2003 values). Clinical data were taken from previously published trials. A 3% discount rate was applied to both resources and life-years gained. RESULTS: Based on model estimates, mean survival times with the third-generation aromatase inhibitors anastrozole and letrozole were 30.72 and 30.64 months, respectively, as opposed to 27.28 months with tamoxifen. Mean survival times after adjustment for quality of life were 18.84 and 18.78 months with anastrozole and letrozole, respectively, and 16.14 months with tamoxifen. Baseline analysis produced incremental cost-effectiveness ratios per quality-adjusted life-year gained of 10,795 Euro (95% CI, 7737 Euro-12,899 Euro) and 16,886 Euro (95% CI, 9117 Euro 15,465 Euro) for anastrozole and letrozole, respectively, compared with tamoxifen. The observed difference between the 2 cost-utility ratios may have been mainly due to the higher acquisition costs of letrozole compared with anastrozole. Despite similar incremental cost-effectiveness ratios, anastrozole and letrozole might increase the budget for advanced breast cancer care by 12% and 18%, respectively, based on the year-2003 Italian market prices of the 2 drugs. CONCLUSIONS: In this cost-effectiveness analysis using previously published clinical data and year-2003 cost data from a community hospital in the Italian National Health Service, anastrozole and letrozole were both cost effective alternatives to tamoxifen for first-line therapy of postmenopausal women with advanced estrogen receptor-positive breast cancer. PMID- 15531018 TI - Cardiovascular aging and psychometric performances: correlations in a group of ultraseptagenarian elderly. AB - Aging of the Italian population resulted in a net increase of the cardiovascular pathologies, and the correlated disabilities. In addition, the cardiovascular diseases represent actually in Italy the most frequent cause of death. With advancing age, both the heart and the blood vessels undergo numerous morphological and functional modifications, which are reducing the functional reserves of these organs. The present study looked for correlation between the cardiac functionality and the cognitive, as well as affective functions. Furthermore, we evaluated the functional variations of the autonomy and autosufficiency of the same patients. We had 171 enrolled subjects (108 women and 63 men), all above the age of 70 years. Based on the classification of the New York Heart Association (NYHA), 85 of these patients (35 men and 50 women) had a II class (Group A), and 86 of them (28 men and 58 women) a III NYHA class of heart function (Group B). We included only patients who did not have any cerebrovascular event yet, and were not bed-ridden. The psychometric performance has been evaluated by using the mini-mental state examination (MMSE), the geriatric depression scale (GDS), the activities of daily living (ADL) and the instrumental activities of daily living (IADL) scales. Cardiac functions have been measured by Doppler echocardiography, in M-mode. The Group A (of mean age 71 +/- 3 years) displayed ventricular ejection fraction (VEF) values in average of 43 +/- 4%, MMSE scores 27 +/- 2; GDS scores 14 +/- 3; IADL 6 +/- 1, and ADL = 6, i.e., maintained a complete autosufficiency. The Group B (mean age 74 +/- 4 years) displayed VEF values in average of 26 +/- 3%, MMSE scores 23 +/- 4; GDS scores 22 +/- 3; IADL 4 +/- 2, and ADL = 4 +/- 1, i.e., had a reduced autosufficiency. These results confirm that also the heart pays a toll for aging: the myocardial contractility becomes significantly altered, meaning the loss of cardiac functions itself. These morpho-functional heart alterations are accompanied by decreased psychometric performances during aging, with consequent reductions of cognitivity, affectivity, autosufficiency and autonomy, involving a complex decrease of the quality of life. PMID- 15531019 TI - Correlation of Mini-Mental-State-Examination (MMSE), Syndrom-Kurztest (SKT) and Clock test (CT) scores in patients with cognitive impairment assessed by means of multiple regression and response surface analysis. AB - The objective of our study was to assess the correlation of routine neuropsychological test results in elderly patients referred to a gerontopsychiatric ward. MMSEs, CTs and SKTs were performed in 94 patients (age: median = 74 years, range = 54-89 years; 64 f, 30 m) with mild to moderate dementia and evaluated retrospectively. Pairwise Spearman rank correlation, multiple regression and response surface analysis were used to assess relations between test results. The scores of all three tests used were reciprocally correlated (P < 0.05) yielding the following pairwise R-values: SKT versus MMSE: 0.77, SKT versus CT: 0.69, MMSE versus Clock test: -0.61. Multiple regression analysis showed a maximum correlation of 0.87 and marked standardised beta values, if SKT was chosen as dependent variable. Test scores could be well fitted to both symmetric linear (SKT = a + b x CT + c x MMSE; R(2) = 0.67, P < 0.01) and non-linear (SKT = a + b x CT + c x MMSE + d x CT(2) + e x MMSE(2) + f x CT x MMSE; R(2) = 0.67, P < 0.01) response surfaces. In conclusion, test scores of SKT, CT or MMSE in patients with dementia showed a marked correlation. In clinical practice the may be considered as almost interchangeable test options. PMID- 15531020 TI - Preferences and frequencies of visits to public facilities in old age--a pilot study in a Swedish town center. AB - In societies with an increasing proportion of elderly active people it is valuable to gain knowledge about what elderly people actually do and want to do in relation to activity arenas in the public environment. The main aim of this study was to investigate elderly people's preferences and frequencies of visits to public facilities in a Swedish town center. A further aim was to gather information on elderly people's subjective perception of problematic and favorable environmental conditions in public environments. The study was conducted using a previously developed questionnaire, "My visit preferences to the public environment". The sample comprised 39 persons aged 75-84 years, living in a defined geographic area. The results of this study provide an overview of the public facilities elderly people rate as important to visit. There are public facilities preferred by most elderly people but with varying frequencies of visits. The study also generated information about elderly people's subjective perceptions of problematic and favorable environmental conditions in public environments, demonstrating that more problems were perceived along walking routes in the public outdoor environment than in the public facilities per se. The results are indicative for community planning supporting elderly people's activities in the public environment, but the questionnaire used should be further validated. PMID- 15531021 TI - Physical training in institutionalized elderly people with multiple diagnoses--a controlled pilot study. AB - Reduction in muscle mass and physical function depends on a variety of interacting factors: age, physical activity level, nutritional state and the type and impact of disease. The aim of this study was to investigate the effect of an individualized moderate intensity physical training program on muscle strength, balance, mobility, ambulation and activities of daily living (ADL) in institutionalized elderly people aged 65 and over with multiple diagnoses. Baseline assessments consisted of strength, balance, mobility/ambulation, and ADL. Twenty-one subjects were included in the intervention program. A control group (21 subjects) was first matched in pairs according to gender, age, ADL and mobility, and then by balance, ambulation and strength. The intervention program was individualized and included strength, mobility, balance and endurance training. Follow-up measures were conducted directly after the intervention and 10 weeks later. After drop-out, 20 subjects in the intervention group and 15 subjects in the control group remained for analyses. Balance and mobility improved significantly in the intervention group while declining in the control group. This pilot study indicates that a physical training program may improve functional capacity for institutionalized elderly persons with multiple diagnoses. PMID- 15531022 TI - Socio-demographic factors related to the prevalence of adverse drug reaction at hospital admission in an elderly population. AB - To identify and to measure the association between socio-demographic factors and the prevalence of adverse drug reaction at hospital admission in an elderly population, we carried out a cross-sectional study on a sample of 19,070 patients of the Gruppo Italiano di Farmacoepidemiologia nell'Anziano data base (Italy). The following socio-demographic variables were considered as independent variables: gender, age, education, living arrangement, widowhood/widowerhood, and alcohol and tobacco habit. We estimated the association of these variables with diagnosis of any adverse drug reaction at hospital admission. We used a logistic regression model to obtain estimates of this association, taking into account hepatic and renal diseases, number of diseases, number of drugs used, albumin index (indicator of nutritional condition) and cognitive function. Out of the 19,070 patients included in our study, 878 (4.3%) were diagnosed with an adverse drug reaction at admission. The multivariate analysis shows that none of the socio-demographic variables and health habits considered are associated with adverse drug reactions at admission. A worse nutritional condition of the subject (OR = 1.39; 95% CI: 1.17-1.64), the number of drugs consumed in the month previous to the admission (OR = 1.74; 95% CI: 1.47-2.08) and the presence of renal disease (OR = 1.76; 95% CI: 1.41-2.55) are associated with ADR at hospital admission. Our results suggest that special attention should be paid to avoid adverse drug reactions in elderly people with nutritional problems, renal failure or those who are using two drugs or more. Additional precautions should be taken with people with cognitive impairment, because of their reduced capacity to report their symptoms. PMID- 15531023 TI - The prevalence of Vitamin D deficiency and effects of ultraviolet light on Vitamin D levels in elderly Turkish population. AB - Vitamin D deficiency is commonly found in the elderly and is associated with osteoporosis and hip fractures. In this study, Vitamin D status of 138 female and 87 male subjects living in old age homes (OAH) and 171 female and 24 male subjects living in own homes (OH) from Central Anatolia were assessed. A questionnaire was applied to collect information about wearing features and degree of sunlight exposure and benefiting from ultraviolet index calculated (BFUI). We have found Vitamin D deficiency in 33.4% among our subjects. Also, 40.1% of subjects living in OAH (54.1% of females and 18.4% of males) and 24.4% of subjects living in OH (27.9% of females and 4.2% of males) were Vitamin D deficient. Vitamin D deficiency was significantly higher in subjects living in OAH than subjects living in OH (P = 0.001) and also higher in females than males (40.7% versus 15.3%, P < 0.001). Subjects with Vitamin D deficiency were older (P < 0.001), BFUI was lower (P < 0.001) and parathyroid hormone (PTH) was higher (P < 0.001) than those having normal level of 25(OH)D. There was a significant negative correlation between 25(OH)D levels and age (P < 0.001, r = -0.248) PTH and 25(OH)D (P = 0.004, r = -0.340), and positive correlation between 25(OH)D and BFUI (P < 0.001, r = 0.340). Vitamin D deficiency is very common in Turkish elderly subjects especially living in OAH and there is a significant low exposure to sunlight among them. Simply by asking clothing habits and exposure to sunlight, we can able to identify risk of Vitamin D insufficiency in elderly subjects. PMID- 15531025 TI - Cost-effectiveness of long-term care services in South Korea. AB - This study examines the economic validity of home health-care and nursing-home care with the aim of providing guidelines for efficient use of limited health care resources. Data collection took place over 8 months in 2001 (from May to December) at six institutions: two home health-care service providers and four nursing homes. A total of 99 stroke patients (49 from home health-care service providers and 50 from nursing homes) participated in the study. The findings indicate that patients with a lower level of physical/cognitive dependency (activities of daily living (ADL) >or= 9.3, cognitive performance scale (CPS) >or= 3.3) tend to benefit more from home health-care service, while those with a high dependency level (ADL < 9.3, CPS < 3.3) receive more suitable care at nursing homes. The study confirms that the economic value of health-care providers varies with the level of physical/cognitive function of the patients. That is, higher efficiency is achieved when those with a lower and higher levels of dependency are provided with home health-care services and nursing-home care, respectively. When assigning long-term care services, it is suggested that the level of physical/cognitive function of patients should be taken into consideration. PMID- 15531024 TI - Non-invasive evaluation of endothelial function in hypertensive elderly patients. AB - Impaired endothelium-dependent vasomotion is a diffuse disease process resulting in abnormal regulation of blood vessel tone and loss of several atheroprotective effects of the normal endothelium. The aim of the present study was to investigate the effects of aging and hypertension on endothelial function. Sixty six geriatric subjects with ages over 60 (48 hypertensive and 18 healthy) and 40 middle-aged subjects (16 hypertensive and 24 healthy) were included in the study. Systemic vascular endothelial function was evaluated through measuring brachial arterial vasodilation, a physiologic answer to reactive hyperemia occured with increased blood flow in the vessel after transient ischemia (flow-mediated dilation, FMD%), and with carotid artery intima-media thickness (IMT) measurement, using high-resolution ultrasonography. Endothelial independent vasodilation was also measured after administration of sublingual isosorbide dinitrate (isosorbide dinitrate mediated dilation, IDNMD%). FMD% was significantly decreased in elderly and/or hypertensive (HT) patients (geriatric HT: 9.5 +/- 4.7%, geriatric non-HT: 12.7 +/- 5.5%, middle-aged HT: 12.9 +/- 4.3% and middle-aged non-HT: 18.9 +/- 8.1%) (geriatric HT versus geriatric non-HT (P = 0.02), geriatric HT versus middle-aged HT (P = 0.01), geriatric non-HT versus middle-aged non-HT (P = 0.008)). Both FMD% and IDNMD% were inversely correlated with age, baseline vessel diameter and carotid artery intima-media thickness. FMD% was also inversely correlated with diastolic blood pressure. No correlation was found between FMD% and systolic blood pressure, serum cholesterol and triglyceride levels. Endothelium dependent (EDD) and independent dilatation of large arteries decreased with aging even in the healthy elderly, and FMD further declined in HT elderly patients, indicating that age and hypertension independently impair endothelial function. Positive correlations with age and hypertension, and significant inverse correlation with FMD, makes carotid artery IMT a possible indicator of endothelial function. PMID- 15531026 TI - Patterns of health complaints among people 75+ in relation to quality of life and need of help. AB - This cross-sectional study aimed to investigate patterns, type and degree of health complaints and their relations to need of help and health-related quality of life (QoL) across gender among people aged 75-105 (n = 4277, mean age 83.6, S.D. 5.7) who answered a postal questionnaire covering health complaints, self reported diseases socio-economy, QoL and need of help with personal and instrumental activities of daily living (PADL, IADL). A principal component analysis gave six categories of health complaints, of which communication (80.9%), mobility problems (66.6%), and psychosocial problems (61%) were most prevalent followed by elimination (42.5%), respiratory-circulatory (38.2%) and digestion-related problems (36.4%). Women reported significantly lower QoL than men. Those needing help with PADL and IADL had significantly lower QoL than those not requiring help. Mobility problems were the strongest predictor besides age, socio-economic factors and female gender for need of help with PADL, IADL and low physical QoL (OR 3.97, 3.67 and 7.47 respectively). Psychosocial problems (OR 3.60) were the strongest predictor besides age, socio-economic factors and female gender for low mental QoL. The findings indicate the importance of focusing on health complaints in coexistence patterns described as dysfunctions, and also the need for primary and secondary preventive actions related especially to mobility and psychosocial problems in geriatric care. PMID- 15531027 TI - Nocturnal giddiness in relation to nocturia and other symptoms and to medication in the elderly. AB - The aim of this study was to evaluate the relation of nocturia, somatic diseases, symptoms, and medication to nocturnal giddiness in a group of elderly men and women. A questionnaire survey was undertaken among 10,216 elderly subjects. The mean (+/-S.D.) ages of the men and women were 73.0 +/- 6.0 and 72.6 +/- 6.7 years, respectively. Nocturnal giddiness was reported by 14.1% of the men and 9.1% of the women. Poor health was reported by 44.4% (P < 0.0001) of the men with nocturnal giddiness and by 14.0% of the men without, and among the women these figures were 45.5% and 20.0% (P < 0.0001), respectively. In a multiple logistic regression analysis significant independent correlates of nocturnal giddiness were: nocturnal micturition episodes >or=3 versus nocturia or=80 years versus <70 years (2.5; 1.7-3.6); eyesight, poor versus good (1.8; 1.4-2.4); hearing, poor versus good (1.4; 1.1-1.9); pain in the cervical spine (2.1; 1.5-2.8); spasmodic chest pain (1.5; 1.1-2.0); diabetes (1.6; 1.0 2.4); analgesics (1.8; 1.3-2.4); and diuretics (1.4; 1.1-21.8). Sex, irregular heartbeats, and sleep medication were deleted by the logistic model. PMID- 15531028 TI - Self-assessment of immediate post-vaccination pain after two different MMR vaccines administered as a second dose in 4- to 6-year-old children. AB - The aim of this study was to compare self-reported immediate post-vaccination pain and safety of Priorix versus RORVax in 4- to 6-year-old children receiving their second dose of MMR vaccine, using the Faces Pain Scale-Revised (FPS-R), a validated self-report pain scale recommended by the French National Accreditation Agency and Health Evaluation. A total of 620 children from 28 French pediatricians completed all study procedures. Immediate post-vaccination pain was reported by 17.8% of the subjects in Priorix group (N=309) and by 44.7% of the subjects in RORVax group (N=311) [OR=3.7; P<0.001]. Parents' pain scores correlated significantly with children's scores. The reduction of immediate pain incidence in Priorix group persisted over the 4 post-vaccination days. This study, using a validated self-assessment pain scale, confirmed previous data showing a significantly lower incidence of immediate post-vaccination pain with Priorix as compared to RORVax. PMID- 15531029 TI - Effectiveness of a publicly funded pneumococcal vaccination program against invasive pneumococcal disease among the elderly in Victoria, Australia. AB - Within Australia, Victoria is the only jurisdiction where the 23-valent polysaccharide pneumococcal vaccine (23vPPV) has been publicly funded for the elderly (aged > or = 65 years). We compared age-specific rates of invasive pneumococcal disease (IPD) for periods before and after implementation of the program, and data from a comparable Australian population that does not have a funded program. Vaccine effectiveness (VE) was estimated using the screening and indirect cohort methods. Compared to the pre-program period, there was a 36% reduction in the reported rates of IPD among persons aged > or = 65 years. Adjusted for under-reporting in the referent rate, the decrease was equivalent to an annual reduction of 112 cases and an estimated 14 deaths among persons > or = 65 years. VE was 71% (95% CI 54-82) using the screening method and 79% (95% CI 14 to 96) by the indirect cohort method. Both point estimates were consistent with the VE expected among persons aged > or = 65 years, although the small number of isolates meant the indirect cohort method was inconclusive at the lower 95% confidence limit. Consideration should be given to publicly funding pneumococcal vaccine for this age group in other settings. PMID- 15531030 TI - SHIV virus-like particles bind and activate human dendritic cells. AB - We previously demonstrated that mucosal immunization with SHIV virus-like particles (VLPs) was able to induce strong humoral and cellular immune responses against HIV envelope protein (Env). To understand the mechanism for such enhanced immune responses, we studied the interaction between VLPs and dendritic cells (DCs) in initiating immune responses. We found that about 50% of DCs were bound by octadecyle rhodamine B (R18) labeled SHIV VLPs. The bound SHIV VLPs were internalized by DCs when cultured at 37 degrees C. Incubation of immature human PBMC-derived DCs with SHIV VLPs for 48 h resulted in the significant up regulation of CD40, CD80, CD83, CD54, CD86, HLA-A, B, C and HLA-DR, DP, DQ molecules on activated DC CD11c+ subpopulations. SHIV VLPs efficiently stimulated DCs to release IL-12, IFN-gamma and TNF-alpha. Furthermore, SHIV VLPs-activated DCs were fully functional in inducing allogeneic T cell proliferation. We conclude that DCs can interact and process SHIV VLPs efficiently and may be critical in initiation of SHIV VLPs-induced immune responses. Thus, interaction between VLPs and DCs may play an important role in the enhancement of immune responses in VLPs-based vaccination. PMID- 15531031 TI - Poly-guanosine strings improve cellular uptake and stimulatory activity of phosphodiester CpG oligonucleotides in human leukocytes. AB - CpG-oligonucleotides (CpG-ODN) have been shown to exert strong immuno-stimulatory effects through activation of Toll-like receptor 9 (TLR-9). However, TLR-9 triggering takes place in endosomal compartments and thus CpG-ODN have to be taken up prior to signal transduction. We here report that 3'-poly-guanosine strings can improve cellular internalisation of phosphodiester but not of phosphorothioate CpG-ODN. Improved cellular uptake correlated with enhanced IL-6 secretion and proliferation of PBMC. Also, TLR-9 transfected HEK293 cells were activated more efficiently by poly-guanosine modified CpG-ODN. The results indicate that the synthesis of stimulatory CpG-ODN based on a phosphodiester backbone is feasible via such poly-guanosine substitutions. In addition we observed that phosphorothioate ODN were able to exert immunostimulatory effects independent of the presence of CpG motifs. PMID- 15531032 TI - Safety of the yellow fever vaccine during the September 2001 mass vaccination campaign in Abidjan, Ivory Coast. AB - In 2001, a vaccination campaign against yellow fever was carried out in Abidjan, Cote d'Ivoire. During the campaign and 4 weeks after an active surveillance system for adverse events following immunization (AEFI) was set up. More then 2.6 million doses were administered and 87 AEFI were notified. Eight suspected YF cases were reported after vaccination and considered as AEFI. However, none had IgM for YF and all recovered without sequels. This surveillance system provided reassuring data about the safety of the YF vaccine and proved that it is feasible to set up an active surveillance system during a mass campaign. PMID- 15531033 TI - Construction and characterization of a Fab recombinant protein for Japanese encephalitis virus neutralization. AB - Filamentous phage display systems have been developed successfully to generate functional Fab antibody fragments. In this study, a recombinant Fab antibody fragment was successfully cloned from a murine monoclonal antibody 2H2 that can effectively neutralize Japanese encephalitis virus (JEV) in vitro. The recombinant Fab 2H2 antibody fragment expressed in Escherichia coli using the pComb3H phage vector resulted in a dose-dependent neutralization response using plaque reduction neutralization test. Molecular modeling of the Fab 2H2 indicated that the rational contact residues of the Fab 2H2 were targeted to the lateral surface of domain III of the JEV E protein. The combining sites of Fab 2H2 were mostly located at the variable region of the heavy chain genes. In vitro shuffling of the heavy-chain variable genes using pCom3H phage technology indicated that the sequence analysis of 10 high-affinity clones selected from the self-shuffling libraries presented no change in their amino acid sequences in 6CDRs, suggesting that the Fab 2H2 had evolved to be highly matured in the combining sites to the lateral surface of domain III. The information gained from this study may benefit the design of vaccines and therapeutic antibodies against JEV infection. PMID- 15531034 TI - Phase 1 study of HPV16-specific immunotherapy with E6E7 fusion protein and ISCOMATRIX adjuvant in women with cervical intraepithelial neoplasia. AB - PURPOSE: Persistent infection of cervical epithelium with "high risk" human papillomavirus (HPV) results in cervical intraepithelial neoplasia (CIN) from which squamous cancer of the cervix can arise. A study was undertaken to evaluate the safety and immunogenicity of an HPV16 immunotherapeutic consisting of a mixture of HPV16 E6E7 fusion protein and ISCOMATRIX adjuvant (HPV16 Immunotherapeutic) for patients with CIN. EXPERIMENTAL DESIGN: Patients with CIN (n = 31) were recruited to a randomised blinded placebo controlled dose ranging study of immunotherapy. RESULTS: Immunotherapy was well tolerated. Immunised subjects developed HPV16 E6E7 specific immunity. Antibody, delayed type hypersensitivity, in vitro cytokine release, and CD8 T cell responses to E6 and E7 proteins were each significantly greater in the immunised subjects than in placebo recipients. Loss of HPV16 DNA from the cervix was observed in some vaccine and placebo recipients. CONCLUSIONS: The HPV16 Immunotherapeutic comprising HPV16E6E7 fusion protein and ISCOMATRIX adjuvant is safe and induces vaccine antigen specific cell mediated immunity. PMID- 15531035 TI - Influenza in Thailand: a case study for middle income countries. AB - Recent studies in Hong Kong and Singapore suggest that the annual impact of influenza in these wealthy tropical cities may be substantial, but little is known about the burden in middle-income tropical countries. We reviewed the status of influenza surveillance, vaccination, research, and policy in Thailand as of January 2004. From 1993 to 2002, 64-91 cases of clinically diagnosed influenza were reported per 100,000 persons per year. Influenza viruses were isolated in 34% of 4305 specimens submitted to the national influenza laboratory. Vaccine distribution figures suggest that less than 1% of the population is immunized against influenza each year. In January 2004, Thailand reported its first documented outbreak of influenza A H5N1 infection in poultry and the country's first human cases of avian influenza. Thailand's growing economy, well developed public health infrastructure, and effective national immunization program could enable the country to take more active steps towards influenza control. PMID- 15531036 TI - Evaluation in macaques of HIV-1 DNA vaccines containing primate CpG motifs and fowlpoxvirus vaccines co-expressing IFNgamma or IL-12. AB - Induction of HIV-specific T-cell responses by vaccines may facilitate efficient control of HIV. Plasmid DNA vaccines and recombinant fowlpoxvirus (rFPV) vaccines are promising HIV-1 vaccine candidates, although either vaccine alone may be insufficient to protect against HIV-1. A consecutive immunisation strategy involving priming with DNA and boosting with rFPV vaccines encoding multiple common HIV-1 antigens was further evaluated in 30 macaques. The DNA vaccine vector included CpG immunostimulatory molecules, and rFPV vaccines were compared with rFPV vaccines co-expressing the pro-T cell cytokines IFNgamma or IL-12. Vaccines expressed multiple HIV-1 genes, mutated to remove active sites of the HIV proteins. The vaccines were well tolerated, and a significant enhancement of DNA-vaccine primed HIV-1 specific T lymphocyte responses was observed following rFPV boosting. Co-expression of IFNgamma or IL-12 by the rFPV vaccines did not further enhance immune responses. Non-sterilising protection from a non pathogenic HIV-1 challenge was observed. This study provides evidence of a safe, optimised, strategy for the generation of T-cell mediated immunity to HIV-1. PMID- 15531037 TI - Limitations of plasmid vaccines to complex viruses: selected myxoma virus antigens as DNA vaccines were not protective. AB - Myxoma virus, a poxvirus of the genus Leporipoxvirus, is the causative agent of the disease myxomatosis which is highly lethal in European rabbits (Oryctolagus cuniculus). Current vaccines to protect against myxomatosis are either attenuated live strains of the virus or the antigenically related rabbit fibroma virus. We examined the immune response of outbred domestic rabbits to the individual myxoma virus antigens M055R, M073R, M115L and M121R, delivered as DNA vaccines co expressing rabbit interleukin-2 or interleukin-4. M115L and M121R were also delivered simultaneously. None of the vaccine constructs were able to protect the rabbits from disease or reduce mortality after challenge with virulent myxoma virus, despite induction of antigen-specific cell-mediated and humoral immune responses. PMID- 15531038 TI - Antibody responses to intradermal recombinant hepatitis B immunization among HIV positive subjects. AB - We conducted a randomized, controlled clinical trial to determine the immunogenicity of intradermal immunization with recombinant hepatitis B vaccine among HIV-positive subjects. Induction of antibody concentration over 10 IU/L or four-fold increase in the antibody concentration against hepatitis B surface antigen was regarded a successful immunization. Intradermal immunization induced protective immunity in 39% of participants who received three doses of recombinant hepatitis B vaccine. Intradermal immunization may provide a way to improve the outcome of hepatitis B vaccination among HIV-infected persons. Three doses of intradermal immunization alone induces protective immunity against hepatitis B as often as intramuscular immunization. PMID- 15531039 TI - Sudden rise in uptake of hepatitis B vaccination among injecting drug users associated with a universal vaccine programme in prisons. AB - Hitherto, services have failed to deliver the UK Government's 1988 recommendation to vaccinate injecting drug users (IDUs) against hepatitis B virus (HBV). In April 1999, the Scottish Prison Service implemented an initiative to offer HBV vaccination to all inmates; we sought to determine the impact of this initiative on the IDU population. Among community-recruited IDUs (who had injected for < or =5 years) in Glasgow, vaccine uptake was significantly higher among those surveyed in 2001-2002 (52% of 387) than in 1993 (16% of 166), 1994 (19% of 138) or January-March 1999 (15% of 128); of the 2001-2002 vaccinees, 56% had been vaccinated in prison. Our results indicate that the universal offer of vaccination to all prisoners, within two years of the initiative's implementation, has had a dramatic impact on uptake among IDUs. PMID- 15531040 TI - Protective and disease-enhancing immune responses induced by recombinant modified vaccinia Ankara (MVA) expressing respiratory syncytial virus proteins. AB - Modified vaccinia Ankara (MVA) recombinants expressing single or multiple RSV surface proteins (F or G) are promising potential vaccines. We studied humoral and cellular responses induced by MVA-F and MVA-G in mice, comparing them to a formalin inactivated RSV preparation (FI-RSV) known to increase disease severity. MVA-F or MVA-G vaccination enhanced weight loss during RSV challenge, but did not show the lung eosinophilia seen after FI-RSV vaccination. FI-RSV induced a stronger total RSV IgG response than the MVA recombinants, but very little IgG2a. MVA recombinants induced cytokine responses biased towards IFNgamma and IL-12, while FI-RSV induced strong IL-4/5 responses in the lungs during RSV challenge. Thus, MVA vaccines induce a favourable immune profile in RSV disease but retain the potential to enhance disease. PMID- 15531041 TI - Plant-synthesized E. coli CFA/I fimbrial protein protects Caco-2 cells from bacterial attachment. AB - A DNA fragment encoding the cholera toxin A2 subunit (CTA2) linked to the enterotoxigenic Escherichia coli (ETEC) colony forming fimbrial antigen CFA/I was inserted into a plant expression vector containing the cholera toxin B subunit (CTB) fused to the rotavirus enterotoxin 22 amino acid epitope NSP422. Anti-CFA/I antibodies recognized a single band of approximately 72-kDa in transformed potato tuber tissue consistent with CFA/I-CTA2 and CTB-NSP4 fusion protein assembly into a cholera holotoxin-like structure. Enzyme-linked immunosorbent assay (GM1 ELISA) indicated that the CFA/I-CTA2 fusion protein bound specific GM1 ganglioside membrane receptors and made up approximately 0.002% of the total soluble tuber protein. Oral immunization of BALB/c mice with transformed tuber tissues generated anti-CFA/I serum and intestinal IgG and IgA secretory antibodies. Attachment of ETEC H10407 to enterocyte-like Caco-2 human colon carcinoma cells incubated with antiserum from immunized mice was reduced by 15% in comparison with Caco-2 cells incubated with serum from unimmunized mice. Immunogold staining of bacterial preparations revealed deposition of gold particles on E. coli H10407 fimbria incubated with immune serum but not on fimbria treated with sera from unimmunized mice demonstrating the specificity of antibodies in the immune serum for binding to CFA/I protein containing fimbria. The protection against toxic E. coli binding to Caco-2 cells generated by antisera from mice immunized with plant synthesized CFA/I antigen demonstrates the feasibility of plant-based multi component vaccine protection against enterotoxigenic E. coli, rotavirus and cholera, three enteric diseases that together exert the highest levels of child morbidity and mortality in economically emerging countries. PMID- 15531042 TI - Effect of egg yolk antibody on experimental Cryptosporidium parvum infection in scid mice. AB - In this study the effect of chicken egg yolk antibody (IgY) against Cryptosporidium parvum infection was examined. IgY sample was prepared from eggs of chickens immunized with C. parvum oocyst antigens. In vitro, antibody-treated sporozoites showed reduced binding to Caco-2 cells and lost vitality. These phenomena were not observed with a control IgY sample prepared from eggs of non immunized chickens. Scid mice orally administered with the antibody demonstrated partial reduction in oocyst shedding after challenge with 10(3) oocysts. IgY, however, could not eliminate the infection after 17 days of continuous treatment. The potentials of using specific IgY for treatment and prevention of cryptosporidiosis were discussed. PMID- 15531043 TI - Interleukin-12 redirects murine immune responses to soluble or aluminum phosphate adsorbed HSV-2 glycoprotein D towards Th1 and CD4+ CTL responses. AB - The type of immune response elicited against HSV-2 infection may be a factor in the frequency and severity of recurrent disease, with non-recurrent status being associated with a Th1-like response. As administration of glycoprotein D subunit formulated with an aluminum-based adjuvant induces predominantly Th2-like immune responses, we sought to assess the ability of IL-12 to redirect anti-HSV immunity towards a Th1 response. Co-administration of gD with IL-12 resulted in gD specific antibody subclass switching from predominantly IgG1 observed in mice immunized with either gD or gD/AlPO4 to a more balanced combination of IgG1 and IgG2a, and enhanced virus neutralizing activity. Spleen cells from mice immunized with gD and IL-12, and restimulated in vitro with HSV-2, developed into effector cells capable of secreting IFN-gamma and lysing HSV-2 infected targets, while those obtained from gD or gD/ALPO4 immunized mice did not express lytic activity. In vitro studies determined that these CTLs were CD4+ and that the cytotoxicity was primarily perforin dependent. Vaginal challenge with HSV-2 demonstrated that IL-12 co-administration with gD resulted in increased efficacy of this vaccine as compared to administration of gD antigen alone. This acquired protection persisted up to 1 year. Finally, adsorbing gD and IL-12 to AlPO4 decreased the optimal dose of IL-12 required to enhance gD immunogenicity and shift responses towards a Th1-like profile. PMID- 15531044 TI - Life-long systemic protection in mice vaccinated with L. major and adenovirus IL 12 vector requires active infection, macrophages and intact lymph nodes. AB - Immunization with soluble leishmanial antigen (SLA) in IFA plus Ad5IL-12 vector induced protection confined to the immunized footpad in BALB/c mice. However, animals that controlled a primary infection with a Leishmania major challenge in the same immunized footpad, became resistant to subsequent contralateral rechallenges due to expansion of IFN-gamma secreting cells. This systemic immunity could be disrupted either by macrophage depletion during immunization or by lymphadenectomy after challenge. We show that this procedure does not interfere with tissue-compartmentalized protection, since lymphadenectomized and splenectomized animals were resistant to rechallenges performed in the immunized footpads. Our results indicate that SLA-Ad5IL-12 vector priming requires macrophages to generate systemic protection. Furthermore, a previously undescribed lymphoid organ-independent, protective immune response is contained within the tissue microenvironment of the immunized/challenged footpad. These results have important implications for vaccine design against leishmanial and mycobacterial infections and diseases caused by intracellular pathogens. PMID- 15531045 TI - Short peptide sequences containing MHC class I and/or class II epitopes linked to nano-beads induce strong immunity and inhibition of growth of antigen-specific tumour challenge in mice. AB - Peptide based vaccines offer practical advantages, but unmodified peptides usually require an adjuvant or delivery vehicle to promote immunogenicity. When peptides containing ovalbumin (OVA) derived CD4 and CD8 T cell epitopes were conjugated to 0.05 microm nano-beads, they gave strong immune responses and inhibition of growth of tumour cells expressing the CD8 T cell epitope with MHC class I. These responses were inducible with both high (50 microg) and low (5 microg) peptide doses after a single immunisation. The helper CD4 T cell epitope was unnecessary for induction of CD8 T cell or tumour challenge responses. However, the CD4 T cell epitope contained a B cell epitope and triggered strong antibody responses. This simple approach offers a convenient experimental tool and a potentially useful clinical method for peptide immunisation. PMID- 15531046 TI - Immunogenicity of polyepitope libraries assembled by epitope shuffling: an approach to the development of chimeric gene vaccination against malaria. AB - Developing a polyepitope vaccine which contains diverse antigenic types is a promising strategy to cope with the problem of malaria variation and diversity. However, arranging the peptides to produce the most effective immunogenicity remains a hurdle. In an attempt to develop an effective complex antigenic gene vaccine, we constructed a polyepitope library by randomly assembling epitopes using the epitope shuffling technique. The polyepitope library, which contains epitopes from different antigens of Plasmodium falciparum, was divided into five sub-libraries based on the size of chimeric genes. Here we report that higher antibody titers were observed in mice with immunized with sub-libraries containing genes >1200 bp, using enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IFAT) assay to determine both individual epitope peptides and the natural parasites at the erythrocyte stage. Different levels of IgG subtypes and cytokines were elicited by different sub-library and administration times. In a rodent malaria model, some groups of immunized mice were partially cross-protected against a lethal challenge from Plasmodium yoelii. These results suggest that the immunogenicity of a polyepitope chimeric antigen is essentially conformation- and length-dependent, and demonstrates that the promising advantage of epitope shuffling technology is that it allows us to randomly assemble many polyepitope molecules in tandem format. This finding also indicates that polyepitope library vaccination is a suitable approach for screening optimized chimeric gene vaccines against malaria and other diseases. PMID- 15531047 TI - Lower extremity joint coupling during running: a current update. AB - BACKGROUND: The relationship between lower extremity mechanics and injury is not well understood. However, joint coupling studies are beginning to emerge, which may lend further insight into running biomechanics. PURPOSE: To provide a current review of the research examining lower extremity joint coupling in running. SUMMARY: There are various techniques utilized to measure joint coupling, including joint timing, rearfoot eversion/tibial internal rotation ratios, continuous relative phase calculations, and vector coding. The study of joint coupling is of particular interest as it may pertain to running injuries. There is some evidence that joint coupling may be altered with orthotics and/or with footwear. Most studies have included a relatively small sample size and larger scale studies are needed to quantify normal ranges for many of the coupling measures. In addition, prospective studies are needed to clarify the relationship to injury. RELEVANCE: It is hoped that this update will serve as a review of the current state of thought regarding lower extremity joint coupling during running. As greater insight into the role of joint coupling in injuries is gained, more optimal intervention and prevention strategies can be developed to minimize injury risk. PMID- 15531048 TI - Differences in motor recruitment and resulting kinematics between low back pain patients and asymptomatic participants during lifting exertions. AB - BACKGROUND: Low back disorders are a prevalent problem in society today and may lead to chronic debilitating low back pain. Developing our understanding of temporal muscle and kinematic patterns during manual material handling tasks may provide insight for preventing the cascading series of events leading to chronic low back pain. METHODS: Sixty-two low back pain patients and 61 asymptomatic participants performed a variety of lifting exertions that varied in lift origin horizontal and vertical distance, lift asymmetry, and weight. Electromyographic activity of 10 trunk muscles as well as trunk and pelvic kinematics was recorded during each exertion. Differences in muscle activation and kinematic parameters were compared between low back pain patients and asymptomatic participants as a function of experimental conditions. FINDINGS: Both the left and right erector spinae activated significantly earlier and were on significantly longer in low back pain patients compared to asymptomatic participants. The horizontal and vertical location of the lift influenced the EMG and kinematic differences between the low back pain patients and asymptomatic participants. INTERPRETATION: These finding indicate that low back pain patients would be exposed to increase muscle activity resulting in higher spine loads for a greater length of time compared to asymptomatic participants. The longer exposure time to increased spine load may lead to greater risk of future low back injury and cascading events leading to debilitating low back pain. The longer muscle activation time suggests that low back pain patients have changed their motor program from an open to a closed loop system. PMID- 15531049 TI - Effect of glenoid component inclination on its fixation and humeral head subluxation in total shoulder arthroplasty. AB - OBJECTIVE: To investigate the effect of glenoid component inclination angle on the fixation of cemented glenoid-bone structures and humeral head subluxation. BACKGROUND: Results of shoulder replacements, in terms of glenoid component fixation and joint functionality, are bad and must be improved. Repeated glenoid component tilting, a result of eccentric contact forces harms glenoid component fixation. However, the effect of glenoid component inclination is unknown. METHODS: Keeled glenoid components are cemented into bone substitutes and positioned under inclination angles varying between -4.5 degrees and +4.5 degrees . For each inclination angle 5 glenoid specimens are loaded by a horizontal, constant joint compression force (725 (SD 10) N) and a vertical, superior directed subluxation force (shear force), cyclically varying between 0 and 350 (SD 1) N. After 200,000 load cycles, the upper and lower glenoid component rim displacements are measured during 1500 additional load cycles by custom made displacement sensors. RESULTS: The maximal superior rim-displacement significantly increased and the minimal inferior rim-displacement dramatically decreased for increasing glenoid component inclination angles up to +4.5 degrees . Maximal ratio of subluxation force over compression force significantly decreased for increasing glenoid component inclination. CONCLUSIONS: According to this study, decreasing glenoid component inclination angle decreases glenoid component tilting and humeral head subluxation and the results can be used to improve total shoulder replacements. RELEVANCE: Better understanding of the effect of glenoid component inclination on glenoid component tilting and humeral head subluxation may be useful information for the surgeon to improve results of the total shoulder arthroplasty, when replacing the glenoid surface. PMID- 15531050 TI - Influence of age on scapulo-thoracic orientation. AB - BACKGROUND: Scapular activity during shoulder motion is critical for normal shoulder function. With aging, muscle function deteriorates in almost all people, which may lead to shoulder impingement syndrome. METHODS: Forty-four normal subjects, 23 men and 21 women aged from 16 to 73 years with a mean age 48, were enrolled in the study. Static antero-posterior radiography at both 0 degrees and 90 degrees of abduction were undertaken and correlation between age and scapular orientations were evaluated by Pearson's correlation coefficient test. The purpose of this study is to investigate the relationship between aging and scapulo-thoracic orientation. FINDINGS: Significant correlation coefficients were observed between aging and scapular orientations. With the shoulder at 0 degrees abduction, posterior tilt showed significant negative correlation with aging. At 90 degrees abduction, both posterior tilt and upward rotation angle correlated negatively with aging. The correlation was apparent at 90 degrees abduction. INTERPRETATION: The results indicate that shoulder aging closely relates to changes of scapular orientation, which consist of decreases of the posterior tilt at 0 degrees and 90 degrees abduction and the upward rotation angle at 90 degrees. The current study indicated that one of the effects of aging on the shoulder is a decrease of posterior tilt and upward rotation angle as seen in an abducted position; and that these alterations are similar to the scapular kinematics of shoulder impingement syndrome. By taking this concept into consideration, the effects of aging on shoulder kinematics can be appropriately evaluated. PMID- 15531051 TI - Force tracking system for the assessment of grip force control in patients with neuromuscular diseases. AB - BACKGROUND: The majority of hand functionality assessment methods consist of the maximal voluntary grip force measurement. Additional knowledge on sensory-motor control can be obtained by capturing functional grip force in a time frame. Tracking methods have been successfully used for the assessment of grip force control in stroke patients and patients with Parkinson's disease. METHODS: A novel tracking system for the evaluation of grip force control is presented. The system consists of a grip-measuring device with the end-objects of different shapes which was used as input to a tracking task where the patient applied the grip force according to the visual feedback. The grip force control was assessed in 20 patients with neuromuscular diseases and 9 healthy subjects. The performance of two tracking tasks was analysed in five grips. The ramp-tracking task was designed to assess the grip strength and muscle fatigue. The sinus tracking task was used to evaluate grip force control during periodic muscle activation. FINDINGS: The results suggest that in some patients the disease did not affect their grip force control despite evident muscular weakness. Most patients produced larger tracking errors in precision grip while the healthy subjects showed less significant differences in performance among the grips tested. INTERPRETATION: The current study investigated force control in patients with neuromuscular diseases where detection of small changes in motor performance is important when following the progress of disease. The presented evaluation method can provide additional information on muscle activation and fatigue as compared to traditional grip strength testing. PMID- 15531052 TI - Influence of gender on hip and knee mechanics during a randomly cued cutting maneuver. AB - OBJECTIVE: To investigate gender differences in three-dimensional hip and knee joint mechanics in collegiate athletes during a randomly cued cutting maneuver. DESIGN: Three-dimensional kinematics and kinetics were collected on 24 collegiate soccer players (12 females and 12 males) while each performed the cutting maneuver. In order to create a randomly cued condition, subjects were signaled by a lighted target board that directed them to perform one of three tasks. Hip and knee joint mechanics were compared between genders using one-tailed t-tests. BACKGROUND: Female athletes have an anterior cruciate ligament injury rate that is larger than their male counterparts. Gender differences in hip and knee joint mechanics during a randomly cued cutting maneuver have not been previously reported. METHODS: Five randomly cued cutting trials were included in the analysis. Selected peak hip and knee joint angles and moments were measured during the first 40 degrees of knee flexion across the stance phase. RESULTS: Females demonstrated significantly less peak hip abduction than did males. Otherwise, there were no gender differences in selected peak hip and knee joint kinematics and moments. CONCLUSIONS: Male and female collegiate soccer players demonstrate similar hip and knee joint mechanics while performing a randomly cued cutting maneuver. RELEVANCE: Because it is known that females incur a greater number of anterior cruciate ligament injuries than males, it is of interest to identify gender differences in lower extremity mechanics when performing sport specific tasks. Understanding of these differences will contribute to the development of prevention training programs. PMID- 15531053 TI - Kinematic analysis of functional lower body perturbations. AB - BACKGROUND: Sudden changes in direction on a single weight-bearing-limb are commonly associated with injury to the lower extremity. The purposes of this study were to assess the between day reliability of hip, knee, and ankle kinematic displacements achieved with internal and external femur-on-weight bearing-tibia rotation perturbations and to determine the effect of these perturbations on three dimensional hip, knee and ankle kinematics. METHODS: Twenty recreationally active, healthy college students with no history of significant orthopedic injury (10 male, 10 female) were subjected to a forward and either internal or external rotary perturbation of the trunk and thigh on the weight-bearing-tibia while three dimensional kinematics were simultaneously collected. The protocol was repeated 24-48 h later to assess reliability. FINDINGS: External perturbations resulted in significant internal rotation (IR) of the tibia on the femur (mean 7.3 (SD 3.9 degrees)) and IR of the femur on the pelvis (mean 6.8 (SD 5.4 degrees)) (P<0.05). Internal perturbations resulted in significant external rotation (ER) of the tibia on the femur (mean 6.8 (5.9 degrees)) and ER of the femur on the pelvis (mean 10.7 (SD 96.1 degrees)) (P<0.05). Additionally the external perturbation results in a significantly greater knee valgus (mean 3.6 (SD 2.2 degrees)) position while the internal perturbation results in a significantly greater knee varus position (mean 2.3 (SD 3.5 degrees)) (P<0.05). External perturbation hip and knee total joint displacements revealed moderate to strong reliability (Intraclass Correlation Coefficient(2,k)=0.67-0.94) while internal perturbations revealed slightly higher Intraclass Correlation Coefficients(2,k)(0.80-0.96). INTERPRETATION: The lower extremity perturbation device provides a consistent external and internal perturbation of the femur on the weight-bearing-tibia. The observed transverse and frontal plane kinematics are similar to motions observed during cross-over and side-stepping tasks. PMID- 15531054 TI - The influence of patella alta on patellofemoral joint stress during normal and fast walking. AB - OBJECTIVE: To determine if persons with patella alta exhibit elevated patellofemoral joint stress compared to pain-free controls during normal and fast walking speeds. SUBJECTS: Twenty-four subjects (13 patella alta, 11 pain-free controls) participated. METHODS: Sagittal and axial magnetic resonance images of the knee were obtained to quantify subject specific knee extensor mechanics and patellofemoral joint contact area. Instrumented gait analysis was used to quantify knee joint kinematics and kinetics. MRI and gait data were used as input variables into a model of patellofemoral joint stress. Analysis of variance with repeated measures was used to compare group differences and group x gait speed interactions for each dependent variable during stance. RESULTS: During normal speed gait there were no group differences in peak knee flexion angle, knee extensor moment, joint reaction force, or stress. However, the patella alta group had significantly less contact area. During fast speed gait there were no group differences in peak knee flexion angle, knee extensor moment, or joint reaction force. However, the patella alta group demonstrated significantly less contact area and significantly greater stress compared to controls. CONCLUSION: Persons with patella alta demonstrated greater calculated patellofemoral stress during fast walking. This was the result of reductions in contact area as joint reaction forces were similar between groups. RELEVANCE: Persons with patella alta may be predisposed to patellofemoral dysfunction through elevations in joint stress. Therefore, treatments aimed at increasing the load-bearing surface area between the patella and femur, such as bracing, may be beneficial in this patient population. PMID- 15531055 TI - Knee ligament behavior following a controlled loading protocol does not differ by menstrual cycle day. AB - BACKGROUND: Females experience a disproportionate number of anterior cruciate ligament injuries compared to males. Increased estradiol concentration has been suggested to alter ligament properties and strength. Determining whether the knee responds differently to an external load at various hormonal levels may be helpful in further explaining the gender disparity. METHODS: Estradiol, progesterone and testosterone were quantified at menses, near ovulation and at the mid-luteal phase. With one knee serving as the control limb and the other as the experimental limb, displacement at 134N and stiffness between 90 and 134N were recorded with a knee ligament arthrometer on both knees before and after a loading protocol. The protocol consisted of three, 3-min, posterior to anterior normalized loads directed to the posterior calf with a ligament testing device. FINDINGS: The loading protocol produced a measurable increase in displacement but not stiffness. Neither displacement nor stiffness measures however were affected by day of the menstrual cycle. No consistent relationships between hormonal concentrations and displacement or stiffness were evident. INTERPRETATION: Following a controlled, static external load, displacement and stiffness were not affected differently by day of the menstrual cycle. PMID- 15531056 TI - The evaluation of plantar pressure distribution in obese and non-obese adults. AB - BACKGROUND: Although previous studies have generally reported increased plantar pressure values with obesity, none of these studies has classified the obesity. Our aim in this study was to compare the plantar pressure distribution in obese and control adults during standing and walking. METHODS: This study was performed on 100 feet of 50 study participants. The subjects gathered in two groups, each containing 25 study participants, as non-obese and class 1 obese according to their body mass index values. Static and dynamic pedobarographic evaluations were performed during standing and walking. The findings were compared between the groups and also the correlation of body mass index with the pedobarographic parameters was assessed. FINDINGS: The static pedobarographic evaluation revealed significantly higher values in terms of forefoot peak pressure, total plantar force and total contact area in the feet of class 1 obese subjects when only middle foot peak pressure was found to be higher in class 1 obese subjects than controls as a dynamic pedobarographic parameter. Among the static parameters body mass index was found to have positive correlation with total plantar force (r=0.50, P=0.000) and total contact area (r=0.33, P=0.019). Only middle foot peak pressure (r=0.32, P=0.025) among the dynamic pedobarographic parameters had positive correlation with body mass index. INTERPRETATION: This study may be a first step to evaluate the effect of different obesity categories on the plantar pressure values. Further studies are needed to investigate the effect of different obesity grades. PMID- 15531057 TI - Two methods of calculating thorax kinematics in children with myelomeningocele. AB - OBJECTIVE: The purpose of this study was to determine the importance of model and cardan angle sequence for calculating thorax kinematics. BACKGROUND: Patients with myelomeningocele often show a significant lateral bending and large trunk rotation over the gait cycle. The temporal pattern and magnitudes of these rotations are important for clinical decision-making. Two common thorax models and two orders of rotation in calculating the thoracic kinematics are compared. METHODS: Thoracic kinematics in eight myelomeningocele patients were studied retrospectively. Four markers were placed on the trunk in addition to the lower limb markers. Each child walked at a self-selected speed over the walkway in bare feet. The spatial position and orientation of each segment were computed from the marker position at 2% of the gait cycle. Cardan angles for two thorax models were calculated using two different orders of rotation with respect to both the laboratory and the pelvic reference systems. RESULTS: The results showed that differences between models in the coronal and transverse planes were greater than in the sagittal plane. The changes in the modelling technique yielded the greatest differences when comparing between models. CONCLUSIONS: Clinical significant differences were found between changes in different thorax models and between the sequences of rotation for subjects with pathological thoracic motion. This clearly indicates that existing thorax models based on indirect marker placement may give misleading kinematic measurements. Different rotation sequences also lead to different results and an analysis of these suggests that the conventional sequence (flexion, lateral bending, axial rotation), is preferable for the thorax. RELEVANCE: Accurate measurement of thoracic motion will enhance our understanding of both the pathological and compensatory mechanism in the gait pattern of children with myelomeningocele. Furthermore, the current investigation on thorax kinematics will lead to calculation of the motion of the center of mass and future methodology to calculate thoracic kinetics. PMID- 15531058 TI - Modifications of baropodograms after transcutaneous electric stimulation of the abductor hallucis muscle in humans standing erect. AB - BACKGROUND: Objective data on abductor hallucis muscle biomechanical function in the loaded foot (subject standing erect on both legs) are unavailable. OBJECTIVE: To evaluate the effects of electrical stimulation of the abductor hallucis muscle in the loaded foot on the change of plantar pressures, as measured by digital baropodograms. DESIGN: Six indices were defined to compare baropodograms. METHODS: The abductor hallucis muscle in 1 foot was subjected to transcutaneous electrical stimulation (20 min) while the subject was standing erect on the floor. Baropodograms were recorded before, immediately thereafter, then 15 days and 2 months later. Differences between baropodogram indices were subjected to one-way anova. RESULTS: Electrical abductor hallucis muscle stimulation induced, on the stimulation side, a post-contraction state easily detected on baropodograms as the increased plantar pressure on the anterior-medial part of the sole, and lateral displacements of the anterior maximal pressure point and the foot thrust center. These mechanical signs, consistent with foot inversion, induce external rotation of the leg and pelvic rotation on the stimulated side, leading to contralateral plantar-pressure changes: decreased maximal pressure point and thrust in the posterior part of the footprint and lateral displacement of the foot thrust center. CONCLUSIONS: Electrical stimulation of the abductor hallucis muscle in the loaded foot induces immediate specific changes in baropodogram indices, some of which persist 2 months later. RELEVANCE: The mechanical effect of abductor hallucis muscle stimulation (foot inversion) and its post-contraction state could be useful in podiatric and postural rehabilitation. PMID- 15531059 TI - Q-angle undervalued? The relationship between Q-angle and medio-lateral position of the patella. AB - BACKGROUND: For an accurate quadriceps angle measurement, the patella must be centralised in the femoral trochlear groove, numerous authors have described lateral displacement of the patella in patellofemoral pain patients, this leads to the intriguing possibility that the Q-angle might be undervalued within patellofemoral pain patients who have laterally displaced patella. METHOD: 109 asymptomatic subjects (51 male, 58 female) were assessed. Medio-lateral patella position was measured using a previous validated method and Q-angle was measured in standing with the quadriceps relaxed. FINDINGS: Mean Q-angle was 11.6 degrees (SD 5.2) left knee, 11.3 degrees (SD 4.9) right knee in the male subjects and 14.4 degrees (SD 5.2) left knee, 13.3 degrees (SD 5.5) right knee for female subjects. 40 females and 28 males had laterally displaced patellae. 13 subjects had centrally placed patellae (7 females, 6 males) with 28 subjects having medially displaced patellae (11 females, 17 males). Recalculation of Q-angle for the laterally displaced group brought about a statistically significant increase in angle. In the medial displaced group failed to produce a statistically significant decrease. INTERPRETATION: The adjusted Q-angle values for medially and neutral placed patellae brought the values very much into the centre of the reported ranges for Q-angles. After adjustment for lateral patella displacement, Q-angle values were towards the under end of values reported as normal, especially female values which were close to the previously reported pathological cut off point. Because of the inverse relationship between quadriceps strength and the magnitude of Q-angle and quadriceps crucial role in the aetiology of patellofemoral pain, any method which improves the reliability and applicability of Q-angle measurement could prove useful in investigations into the aetiology of and outcome from treatment of patellofemoral pain syndrome. PMID- 15531060 TI - Alexander Graham Bell and the vacuum jacket for assisted respiration. PMID- 15531061 TI - International EMS Systems: Canada. PMID- 15531062 TI - Uninterrupted chest compression CPR is easier to perform and remember than standard CPR. AB - INTRODUCTION: It has long been observed that CPR skills rapidly decline regardless of the modality used for teaching or criteria used for testing. Uninterrupted chest compression CPR (UCC-CPR) is a proposed alternative to standard single rescuer CPR (STD-CPR) for laypersons in witnessed unexpected cardiac arrest in adults. It delivers substantially more compressions per minute and may be easier to remember and perform than standard CPR. METHODS: In this prospective study, 28 medical students were taught STD-CPR and UCC-CPR and then were tested on each method at baseline (0), 6, and 18 months after training. The students' performance for at least 90 s of CPR was evaluated based on video and Laerdal Skillreporter Resusci Anne recordings. RESULTS: The mean number of correct chest compressions delivered per minute trended down over time in STD-CPR (23 +/- 3, 19 +/- 4 , and 15 +/- 3; P = 0.09) but stayed the same in UCC-CPR (43 +/- 9, 38 +/- 7, and 37 +/- 7 = 0.91) at 0, 6, and 18 months, respectively. The mean percentage of chest compressions delivered correctly fell over time in STD CPR (54 +/- 6%, 35 +/- 6%, and 32 +/- 6%; P = 0.02) but stayed the same in UCC CPR (34 +/- 5%, 41 +/- 7%, and 38 +/- 8%) at 0, 6, and 18 months, respectively. The number of chest compressions delivered per minute was higher in UCC-CPR at 0, 6, and 18 months (113 versus 44, P < 0.0001; 94 versus 47, P < 0.0001; and 92 versus 44, P < 0.001). The greater number of chest compressions was due to a mean ventilaroty pause of 13-14 s during STD-CPR at all three time points. CONCLUSIONS: Chest compression performance during STD-CPR declined in repeated testing over 18 months whereas there was minimal decline in chest compressions performance on repeated testing of UCC-CPR. In addition, substantially more chest compressions were delivered during UCC-CPR compared to STD-CPR at all time points primarily because of long pauses accompanying rescue breathing. PMID- 15531063 TI - Effects of decreasing peak flow rate on stomach inflation during bag-valve-mask ventilation. AB - Reducing inspiratory flow rate and peak airway pressure may be important in order to minimise the risk of stomach inflation when ventilating an unprotected airway with positive pressure ventilation. This study was designed to yield enough power to determine whether employing an inspiratory gas flow limiting bag-valve device (SMART BAG, O-Two Medical Technologies Inc., Ontario, Canada) would also decrease the likelihood of stomach inflation in an established bench model of a simulated unintubated respiratory arrest patient. The bench model consists of a training lung (lung compliance, 50 ml/cm H2O; airway resistance, 4 cm H2O/l/s) and a valve simulating lower oesophageal sphincter opening at a pressure of 19 cm H(2)O. One hundred and ninety-one emergency medicine physicians were requested to ventilate the manikin utilising a standard single-person technique for 1 min (respiratory rate, 12/min; Vt, 500 ml) with both a standard adult bag-valve-mask and the SMART BAG. The volunteers were blinded to the experimental design of the model until completion of the experimental protocol. The SMART BAG versus standard bag-valve mask resulted in significantly (P < 0.001) lower (mean +/- S.D.) mean airway pressure (14 +/- 2 cm H2O versus 16 +/- 3 cm H2O), respiratory rates (13 +/- 3 breaths per min versus 14 +/- 4 breaths per min), incidence of stomach inflation (4.2% versus 38.7%) and median stomach inflation volumes (351 [range, 18-1211 ml] versus 1426 [20-5882 ml]); lung tidal volumes (538 +/- 97 ml versus 533 +/- 97 ml) were comparable. Inspiratory to expiratory ratios were significantly (P < 0.001) increased (1.7 +/- 0.5 versus 1.5 +/- 0.6). In conclusion, the SMART BAG reduced inspiratory flow, mean airway pressure and both the incidence and actual volume of stomach inflation compared with a standard bag-valve-mask device while maintaining delivered lung tidal volumes and increasing the inspiratory to expiratory ratio. PMID- 15531064 TI - Prediction of short- and long-term outcomes by electrocardiography in survivors of out-of-hospital cardiac arrest. AB - BACKGROUND: Programs focusing on early defibrillation have improved both short- and long-term survival of patients with VF out-of-hospital cardiac arrest (OHCA). Subsequent long-term management of survivors would be facilitated by a straight forward, non-invasive method of identifying those at highest risk for recurrence. Therefore, we assessed the predictive value of the standard ECG to determine both short- and long-term outcomes in survivors of VF OHCA to assist in risk stratification of those patients at highest risk of sudden death. METHODS: All patients with an OHCA between November 1990 and December 2000 who received early defibrillation for VF in Olmsted County Minnesota (MN) were included. Cox proportional hazards modeling was used to examine ECG variables and subsequent ICD deployment and death. RESULTS: Two hundred patients presented in VF OHCA; of these 138 (69%) survived to hospital admission (seven died in the emergency department prior to admission) and 79 (40%) were discharged. The QRS duration (141 +/- 41ms in nonsurvivors, 123 +/- 35 in survivors, P = 0.004) was predictive of short-term mortality in patients who did not survive to hospital discharge. The ventricular rate, PR interval, presence of right or left bundle branch block, QTc, ST elevation myocardial infarction, and atrial fibrillation/flutter were nonpredictive. The average length of follow up for hospital dismissal survivors was 4.8 +/- 3.0 years. In univariate analysis, each 30 ms interval increase in the QRS width and PR interval was associated with increased mortality and ICD deployment hazard ratio of 1.6 (CI 1.1-2.5, P = 0.02) and 1.12 (CI 1.0-1.2, P = 0.05), respectively. In multivariate analysis accounting for admission ejection fraction, a PR > 200 ms [HR 4.5 (CI 1.7-11.8, P = 0.022)], QRS width increase greater than 30 ms [HR 1.9 (CI 1.3-2.8, P < 0.001)], and a QRS > 120 ms [HR 2.4 (CI 1.1-5.4, P = 0.032)] were predictive of long-term mortality and ICD shocks. CONCLUSION: Careful evaluation of the admitting and discharge ECG provides prognostic information for in-hospital and long-term outcomes, respectively in this cohort of out-of-hospital cardiac arrest survivors. The QRS duration on the dismissal ECG following VF OHCA provides prognostic information which might be useful to identify those at highest risk long-term, and who would benefit from more aggressive antiarrhythmic therapy and cardiac stabilization. PMID- 15531065 TI - Prediction rules for estimating neurologic outcome following out-of-hospital cardiac arrest. AB - BACKGROUND: No valid model has been developed to predict survival following out of-hospital cardiac arrest. The purpose of this study was to develop a prediction model for meaningful survival following out-of-hospital cardiac arrest using variables available during resuscitation. METHODS: This was a retrospective cohort study. Consecutive adult cardiac arrest patients were studied between 1994 and 2001. Variables included age, sex, race/ethnicity, arrest location, whether the arrest was witnessed, initial rhythm, whether CPR was performed, patient downtime, paramedic response time, survival to hospital discharge, and Glasgow Coma Score (GCS) at hospital discharge. Classification and Regression Tree analysis was used to develop decision rules to predict meaningful survival, as defined by the patient's discharge GCS. RESULTS: Of the 754 patients, 16 (2%) survived with a GCS > or =13, 15 (2%) survived with a GCS = 14, and 5 (0.7%) survived with a GCS = 15. The decision rule for survival with a GCS > or = 13 incorporated whether the arrest was witnessed and the patient's age, resulting in a negative predictive value (NPV) of 99.8%. The rule for survival with a GCS > or = 14 incorporated the initial arrest rhythm, whether the arrest was witnessed, and the patient's age, resulting in a NPV of 99.6%. The rule for survival with a GCS = 15 incorporated only the interval between collapse and the initiation of life support, resulting in a NPV of 99.8%. CONCLUSIONS: This study reports decision rules for potential meaningful survival following out-of-hospital cardiac arrest with high NPVs for each. Future studies need to be performed to prospectively validate these models. PMID- 15531066 TI - Implementation of cardiopulmonary resuscitation guidelines in elderly care departments across: a survey of 13 hospitals shows wide variability in practice. AB - OBJECTIVE: The United Kingdom Department of Health advises hospitals that they should implement a policy relating to cardiopulmonary resuscitation (CPR) that takes account of published guidelines relating to decision making for resuscitation. We wished to see if these guidelines were leading to implementation of a similar policy in different Elderly Care (EC) departments. SETTING: The acute and rehabilitation wards in 13 hospitals from the South Thames West region. DESIGN: On one day the notes of all in-patients, over 55 years, under the care of an EC physician were reviewed. RESULTS: A CPR decision had been made in 465 (47%) of the 990 in-patients, 379 Do Not Attempt Resuscitation (DNAR) orders were made. The percentage of patients in whom a CPR decision had been made varied from 15 to 97% between departments. CONCLUSIONS: Implementation of the guidelines for decision making for resuscitation varies greatly across this region of the United Kingdom. PMID- 15531067 TI - Three year longitudinal study for out-of-hospital cardiac arrest in Osaka Prefecture. AB - OBJECTIVE: To analyze the longitudinal changes in the treatment of out-of hospital cardiac arrests. These analyses have focused on the time interval from the receipt of call until defibrillation of patients with ventricular fibrillation. DESIGN: Population-based, prospective longitudinal study according to the Utstein style. SETTING: Osaka Prefecture (population 8, 800, 000), served by 36 municipal fire and emergency departments. PATIENTS: Consecutive, out-of hospital cardiac arrests occurring between May 1998 and April 2001. MAIN OUTCOME MEASURES: Change in the interval to defibrillation, and one-year survival from cardiac arrest. RESULTS: Of the 15,211 cases of confirmed cardiac arrests during the three years, resuscitation was attempted in 14,609 subjects. Of the 2957 cases of cardiac origin and witnessed by bystanders, 90 cases (3.0%) were alive 1 year following the episode. In 383 cases of defibrillation, the interval from receipt of call to defibrillation was evaluated annually. This interval decreased significantly during the three year course (14.5, 13.0, and 11.5 min expressed by the median), suggesting that this project to report the data of out-of-hospital arrests was an effective campaign for EMT. However, the outcome did not improve significantly during this period (3.0%, 2.6%, and 3.6% alive 1 year in witnessed arrests with cardiac etiology). This may be because the third year median duration of 11.5 min, is still insufficient to indicate a significant improvement in the outcome. CONCLUSIONS: This project to report the data of out-of-hospital cardiac arrest might have contributed to the reduction of the interval for defibrillation, as a campaign for the EMTs; although the decrease in this interval was still insufficient to result in a significant increase in the number of cases who are alive one year later. PMID- 15531068 TI - Impact of community-wide deployment of biphasic waveform automated external defibrillators on out-of-hospital cardiac arrest in Taipei. AB - OBJECTIVES: To determine the impact and outcome of out-of-hospital cardiac arrests (OHCA) while using automated external defibrillators (AED) with biphasic waveforms and its effectiveness when using the Utstein Style community-wide in Taipei. MATERIAL AND METHODS: A one-year study was conducted to collect OHCA patients with AED utilization prospectively in Taipei City. All events and variables were recorded in the Utstein Style. Electrocardiography and voice records recovered from AED data cards were analysed. The endpoints were survival outcomes. RESULTS: Of 653 OHCA patients with AED utilization, only 80 (12.6%) patients with 635 true arrests presented with ventricular fibrillation or tachycardia (VF/VT) as the initial rhythm. The interval between call-to-shock was 5 min longer than call-to-EMS arrival (9.3 min versus 4.0 min). Fourteen (25%) of the 55 witnessed VF/VT arrests survived to home discharge. Ninety-seven percent of shockable rhythms were successfully terminated with less than three shocks. For all OHCA patients, initial rhythm of VF/VT (OR 3.4; 95% CI = 1.2-9.4), witnessed status (OR 4.7; 95% CI = 1.3-16.6), and presence of organised rhythm irrespective of pulse during prehospital resuscitation (OR 9.2; 95% CI = 3.2 26.8) demonstrated an independent association with survival to home discharge. For VF/VT arrests, witnessed status, shorter call-to-shock time, high successful rate of the first shock, fewer averaged number of shocks delivered for each patient, and presence of an organised rhythm during prehospital resuscitation showed a likelihood to predict to predict discharged survival in univariate analysis. CONCLUSIONS: Low frequency of VF arrests is unique to certain eastern populations but without a reduction of AED shock efficacy with biphasic waveform. Besides initial VF and witnessed status, a prehospital post-shock organized rhythm irrespective of pulse appears to be correlated to survival. Certain circumstances in a congested metropolitan city consume time to deliver shocks even after EMS arrival, and might require bystander or public access defibrillation. PMID- 15531069 TI - Preliminary report on AED deployment on the entire Air France commercial fleet: a joint venture with Paris XII University Training Programme. AB - The positive effect of early defibrillation on survival from cardiac arrest has been demonstrated. We describe the use of AEDs over 1 year following the training of flight attendants. Air France and the University of Paris XII together designed a 1 year training programme for 14000 flight attendants. The university emergency departments (SAMU) provided 250 instructors. AEDs training and certification was conducted for crew members between November 2001 and November 2002. By January 2003, all aircraft were fully equipped with AEDs. All cases of cardiac arrest that occurred during the study were reviewed comprehensively. Comments from the crew were collected. Twelve cardiac arrests were reported between November 2002 and November 2003 out of 4194 cases of emergency care delivered to passengers. Shock treatment was advised initially in 5/12 cases. The survival rate after in-flight cardiac arrest was 3/12. The survival rate at discharge from hospital following in flight shock was 2/5. No complications arose from the use of AEDs. Training by professionals gave the flight attendants confidence and allowed for the survival of two young passengers. Our study highlights the ability of flight attendants to give better onboard care for the future. The next step is to consolidate the network between in-flight care and the medical dispatch centre in Paris. PMID- 15531070 TI - In hospital cardiac arrest: a role for automatic defibrillation. AB - INTRODUCTION: Sudden cardiac death (SCD) survival decreases by 10% for each minute of delay in defibrillation, however, survival rates of 98% can be achieved when defibrillation is accomplished within 30s of collapse. Recently, a fully automated external cardioverter-defibrillator (AECD) was approved by the FDA for in-hospital use. The AECD can be programmed to automatically defibrillate when a life threatening ventricular arrhythmia occurs. The purpose of this study was to assess the potential impact of in-hospital AECDs on the critical time to defibrillation in monitored hospital units. METHODS: Mock emergency (n = 18) were conducted using simulated ventricular fibrillation in various monitored units. Observers were stationed to record the time staff responded to the arrhythmia, and the time to shock. These times were compared to an AECD protocol that defibrillates automatically in an average of 38.3 s from onset of arrhythmia (n = 18). RESULTS: Staff versus AECD response time to arrhythmia (s) was 76.3 +/- 113.7 (CI 19.8-132.8) versus 7.6 +/- 0.6 (CI 7.3-7.9). Staff versus AECD time to shock was 169.2 +/- 103.1 (CI 117.9-220.4) versus 38.3 +/- 0.7 (CI 37.9-38.6). P values are <0.0001 for differences between the groups. CONCLUSION: The use of AECDs on monitored units would significantly reduce the critical time to defibrillation in patients with SCA. We anticipate this would translate to improved survival rates, and better neurologic outcomes. PMID- 15531071 TI - Cerebral oximetry in out-of-hospital cardiac arrest: standard CPR rarely provides detectable hemoglobin-oxygen saturation to the frontal cortex. AB - Favorable neurological survival in out-of-hospital cardiac arrest (OOHCA) may be influenced by cerebral perfusion during resuscitation. Cerebral oximetry (COx) provides a portable, noninvasive, real-time index of cerebral perfusion that has not been studied in OOHCA. This study examined the feasibility of using COx to measure cerebral perfusion during OOHCA. As a secondary aim, we tested the hypothesis that cerebral perfusion, measured by COx, would decrease with hyperventilation. Subjects were patients with medical OOHCA. A physician responded to the scene of cardiac arrest calls and applied an INVOS 3000 COx probe (Somanetics) to the frontal skull. In a cross-over design, readings were recorded for 2 min while ventilation rate was maintained at 10/min, then for 2 min at 24/min. COx readings were recorded by the oximeter and manually by the investigator. Statistical analyses were done using a paired t-test. Sixteen subjects were enrolled, four had return of pulses. COx reliably detected cortical oxygenation in only one subject during cardiac arrest. None of 16 patients exhibited consistently detectable levels of oxygen during cardiac arrest. In three subjects with ROSC, readings increased with return of pulses and with increasing blood pressure. In a fourth subject the protocol was completed and the device removed, subsequently ROSC was noted and the device re-applied. No patient exhibited any change in oxygen levels with variation of ventilation rates during CPR. The use of cerebral oximetry during OOHCA is feasible. In our sample of OOHCA patients, cerebral perfusion is rarely detectable using an oximeter during CPR. Ventilation rate does not alter the oximeter readings. It is possible that the current standard mechanical method of cardiopulmonary resuscitation provides little or no cerebral oxygenation during OOHCA. PMID- 15531072 TI - Etiology of troponin I elevation in patients with congestive heart failure and low clinical suspicion of myocardial infarction. AB - BACKGROUND: Increased troponin I (TnI) has been detected in congestive heart failure (CHF) patients. We demonstrate that occurrence of LVH, and systolic and diastolic blood pressure values (SBP and DBP, respectively) correlate with increasing levels of serum TnI in a selected CHF population. METHODS: Our team reviewed the medical records of 40 consecutive patients admitted with CHF symptoms, low clinical suspicion of myocardial infarction (MI), and serum TnI measured upon admission. Twenty numerical and categorical parameters were screened. Patients were divided in to seven non-exclusive groups by increasing cutoff values of serum TnI (0, 0.1, 0.2, 0.3, 0.4, 0.6, and 1.5 ng/ml). In each group the average value of numerical parameters and the occurrence of categorical ones were calculated. The correlation between these values and the seven TnI cutoffs were analyzed. RESULTS: Occurrence of electrocardiographic left ventricular hypertrophy (ECG-LVH), and average values of SBP and DBP significantly correlated with the TnI cutoffs by means of multiple regression analysis (P = 0.001, 0.02 and 0.007, respectively). CONCLUSION: ECG-LVH and values of SBP and DBP significantly correlate with increasing cutoff levels of serum TnI in hospitalized patients with CHF and low clinical suspicion of MI. PMID- 15531073 TI - Pulmonary transcription of CAT-2 and CAT-2B but not CAT-1 and CAT-2A were upregulated in hemorrhagic shock rats. AB - Hemorrhagic shock stimulates nitric oxide (NO) biosynthesis through upregulation of inducible NO synthase (iNOS) expression. Trans-membrane l-arginine transportation mediated by the isozymes of cationic amino acid transporters (e.g. CAT-1, CAT-2, CAT-2A, and CAT-2B) is one crucial regulatory mechanism that regulates iNOS activity. We sought to assess the effects of hemorrhage and resuscitation on the expression of these regulatory enzymes in hemorrhage stimulated rat lungs. Twenty-four rats were randomized to a sham-instrumented group, a sustained shock group, a shock with blood resuscitation group, or a shock with normal saline resuscitation group. Hemorrhagic shock was induced by withdrawing blood to maintain MAP between 40 and 45mmHg for 60min. Resuscitation by infusing blood/saline mixtures (blood resuscitation group) or saline alone (saline resuscitation group) was then performed. At the end of the experiment (300min after hemorrhage began), rats were sacrificed and enzymes expression as well as pulmonary NO biosynthesis and lung injuries were assayed. Our data revealed that hemorrhage-induced pulmonary iNOS, CAT-2, and CAT-2B transcription which was associated with pulmonary NO overproduction and subsequent lung injury. Resuscitation significantly attenuated the hemorrhage-induced enzyme upregulation, pulmonary NO overproduction, and lung injury. Blood/saline mixtures were superior to saline as a resuscitation solution in treating hemorrhage induced pulmonary NO overproduction and lung injury. Hemorrhage and/or resuscitation, however, did not affect the expression of pulmonary CAT-1 and CAT 2A. It is, therefore, concluded that the expression of pulmonary iNOS, CAT-2, and CAT-2B is inducible and that of CAT-1 and CAT-2A is constitutive in hemorrhagic shock rat lungs. PMID- 15531074 TI - Near-infrared spectroscopy: a tool to monitor cerebral hemodynamic and metabolic changes after cardiac arrest in rats. AB - INTRODUCTION: Cardiac arrest (CA) is associated with poor neurological outcome and is associated with a poor understanding of the cerebral hemodynamic and metabolic changes. The objective of this study was to determine the applicability of near-infrared spectroscopy (NIRS), to observe the changes in cerebral total hemoglobin (T-Hb) reflecting cerebral blood volume, oxygenation state of Hb, oxidized cytochrome oxidase (Cyto-C), and brain water content following CA. METHODS: Fourteen rats were subjected to normothermic (37.5 degrees C) or hypothermic (34 degrees C) CA induced by 8 min of asphyxiation. Animals were resuscitated with ventilation, cardiopulmonary resuscitation (CPR), and epinephrine (adrenaline). Hypothermia was induced before CA. NIRS was applied to the animal head to measure T-Hb with a wavelength of 808 nm (n = 10) and oxygenated/deoxygenated Hb, Cyto-C, and brain water content with wavelengths of 620-1120 nm (n = 4). RESULTS: There were no technical difficulties in applying NIRS to the animal, and the signals were strong and consistent. Normothermic CA caused post-resuscitation hyperemia followed by hypoperfusion determined by the level of T-Hb. Hypothermic CA blunted post-resuscitation hyperemia and resulted in more prominent post-resuscitation hypoperfusion. Both, normothermic and hypothermic CA resulted in a sharp decrease in oxygenated Hb and Cyto-C, and the level of oxygenated Hb was higher in hypothermic CA after resuscitation. There was a rapid increase in brain water signals following CA. Hypothermic CA attenuated increased water signals in normothermic CA following resuscitation. CONCLUSION: NIRS can be applied to monitor cerebral blood volume, oxygenation state of Hb, Cyto-C, and water content following CA in rats. PMID- 15531075 TI - Sub-arachnoid hemorrhage following cardiopulmonary resuscitation. AB - We report the case of a 20-year-old male who developed unexplained sub-arachnoid haemorrhage following cardiopulmonary resuscitation. Computed tomography and lumbar puncture performed within 24 h were normal. A CT performed on day 5 revealed massive sub-arachnoid hemorrhage. The patient expired due to brain herniation. We conclude that sub-arachnoid hemorrhage (SAH) may be a late complication of aborted sudden death. We suggest ischaemia-reperfusion injury as a possible mechanism. PMID- 15531076 TI - Thrombosis after cardiac arrest. PMID- 15531077 TI - Identification of molecular variants at the promoter region of the human alpha 7 neuronal nicotinic acetylcholine receptor subunit gene but lack of association with schizophrenia. AB - The human alpha7 neuronal nicotinic receptor subunit gene has been considered as a candidate gene for P50 sensory gating deficit in schizophrenic patients. Because P50 sensory gating deficit is a common neurophysiological dysfunction in patients with schizophrenia and schizophrenia spectrum disorders, it is conceivable to hypothesize that the human alpha7 neuronal nicotinic receptor subunit gene might be a susceptible gene for schizophrenia. Researchers have reported that mutations in the protein-coding sequences of the human alpha7 neuronal nicotinic receptor subunit gene are very rare. Therefore, we searched for mutations at the promoter region of the human alpha7 neuronal nicotinic receptor subunit gene and performed a genetic association study in 249 unrelated Han Chinese schizophrenic patients and 273 non-psychotic subjects from Taiwan. Two molecular variants were identified and designated g.-213G>A and g.-324A>G, respectively. The g.-213G>A variant was found to obliterate a putative NF-1 transcription factor binding site using computer analysis. One out of 249 patients was detected to be a heterozygote for this variant, but none of 273 control subjects was. The g.-324A>G variant was also very rare in both patients and control subjects, only one heterozygote of this variant was identified in 249 patients and 273 control subjects, respectively. Hence, in this study, we did not find mutations in the human alpha7 neuronal nicotinic receptor subunit gene that are associated with schizophrenia in our population. PMID- 15531079 TI - Depolarization thresholds for hippocampal damage, ischemic preconditioning, and changes in gene expression after global ischemia in the rat. AB - Induced ischemic tolerance in rat hippocampus was investigated in a forebrain ischemia model of repeated 4-vessel occlusion (4-VO). Ischemic insult variability was reduced by the use of dc potential measurements to determine the duration of ischemic depolarization in hippocampus. The results demonstrate a depolarization threshold for ischemic injury to CA1 neurons of 4-6 min and a window for optimal preconditioning of 2.5-3.5 min. Levels of induced mRNAs encoding hsp72 and several immediate-early genes were also shown to vary with depolarization interval. Immediate-early genes were maximally induced after depolarization periods inducing optimal preconditioning, while hsp72 expression increased with insult severity over the range leading to neuron loss. These results are similar to those obtained in gerbil studies indicating that preconditioning does not require large increases in hsp72 expression, and demonstrate the fundamental comparability of rodent global ischemia models when monitored by this approach. PMID- 15531078 TI - Over-expression of the fyn-kinase gene reduces hypnotic sensitivity to ethanol in mice. AB - Our previous work indicated a role for fyn-kinase in mediating several ethanol- and GABA(A) agonist-mediated behaviors. In the present work we investigate behavioral sensitivity to ethanol and several GABA(A) compounds in mice that over express fyn-kinase in forebrain to further characterize the role of this non receptor tyrosine kinase in the mediation of ethanol sensitivity. Transgenic mice over-expressing fyn-kinase were tested for sensitivity to ethanol-induced loss of righting reflex and ethanol preference drinking using a two-bottle choice drinking paradigm. Loss of righting reflex induced by 4,5,6,7 tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP; GABA(A) agonist) and etomidate (GABA(A) positive allosteric modulator) were also assessed. Fyn over-expressing mice exhibited shorter durations of ethanol-induced loss of righting reflex in the absence of differences in the rate of blood ethanol clearance, and exhibited reduced ethanol preference drinking. The genotypes did not differ in initial sensitivity to ethanol-induced loss of righting reflex suggesting development of greater acute tolerance to this ethanol action. Fyn over-expressing and wild-type mice also did not differ in sensitivity to loss of righting reflex induced by THIP and etomidate. The present results suggest regional specificity for fyn kinase in the modulation of ethanol and GABAergic behavioral sensitivity. Fyn kinase over-expression in forebrain structures modulates ethanol's hypnotic actions, as well as ethanol preference and consumption. Moreover, fyn over expression in forebrain does not alter hypnotic sensitivity to THIP or etomidate, supporting data from fyn null mutant mice suggesting that cerebellar structures mediate the hypnotic actions of these GABAergic compounds. PMID- 15531080 TI - The role of Bax-inhibiting peptide in retinal ganglion cell apoptosis after optic nerve transection. AB - Bax, a pro-apoptotic member of Bcl-2 family proteins, plays a central role in mitochondria-dependent apoptosis. Bax normally resides in the cytosol in a quiescent state. Bax-inhibiting peptide (BIP) is a membrane permeable peptide comprised of five amino acids designed from the Bax-binding domain of Ku70 [M. Sawada, P. Hayes, S. Matsuyama, Cytoprotective membrane-permeable peptides designed from the Bax-binding domain of Ku70, Nat. Cell Biol. 5 (2003) 352-357]. It inhibits Bax-mediated translocation of cytochrome c and suppresses mitochondria-dependent apoptosis. BIP was used in order to elucidate its role in preventing retinal ganglion cell (RGC) death from apoptosis after optic nerve transection (ONT) in adult Wistar rats. RGC survival was significantly higher in animals with intravitreal injection of BIP, when compared with control animals. These findings suggest that BIP prevented RGC apoptosis after ONT prompting the suggestion that Bax plays a central role in RGC apoptosis after ONT. PMID- 15531081 TI - Modulation of predatory odor processing following lesions to the dorsal premammillary nucleus. AB - Previous studies have shown that electrolytic lesions of the dorsal premammillary nucleus (PMd) produce robust reductions in responsivity of rats to the presence of a live predator as well as to its odor, suggesting a critical role for the PMd in the modulation of defense. The present study investigated whether disruptions in defensive responding were specific to predators or if they may indicate a more general deficit in responding to pheromonal odors. Sexually naive male rats with bilateral ibotenic acid lesions of the PMd were exposed to the odor of a female rat in estrus as well as to the presence of cat odor, and, a live cat. PMd lesions produced a dramatic reduction in freezing and avoidance to the cat odor; and, reductions in freezing, enhanced activity and risk assessment to cat exposure. However, PMd lesions produced no changes in response to the presentation of the female odorant. These results confirm earlier findings of attenuation in defensiveness following electrolytic PMd lesions while extending these findings to suggest that the reduced defensiveness occurs specifically in response to predatory odors. PMID- 15531082 TI - Association of the HTR6 polymorphism C267T with late-onset Alzheimer's disease in Chinese. AB - Serotonergic neurotransmitter system has been implicated in the pathogenesis of Alzheimer's disease (AD). 5-Hydroxytryptamine (5-HT)(6) receptor is mainly expressed in the brain areas involved in cognitive processes. And 5-HT(6) receptor gene (HTR6) variants may be a genetic risk factor for late-onset Alzheimer's disease (LOAD). To assess whether HTR6 increases the risk for LOAD, we carried out an association study between the HTR6 polymorphism C267T and sporadic LOAD in Mainland Chinese. An association of C/T genotype with LOAD (OR = 2.10, P = 0.014) was observed. And no statistical difference was found between cases and controls after stratification for APOE epsilon4 status. These data suggest that the HTR6 polymorphism C267T possibly involved in the susceptibility to LOAD as an APOE epsilon4-allele independent risk factor of LOAD. PMID- 15531083 TI - Behavioral evidence for competing motivational drives of nociception and hunger. AB - Homeostasis, an organisms' tendency to maintain a healthy balance of the physiological state of the body, is necessary for survival. Hunger induces a motivational state to consume food. Recently, pain has been referred to as a homeostatic emotion similar to hunger or thirst in that animals are motivated to respond in a certain way that may increase their chance of survival. Therefore, the purpose of the present experiment was to examine behavior in rodents during two competing homeostatic/motivational drives (i.e., hunger and formalin pain). During the first phase of the formalin test, animals displayed typical responsiveness to the inflammatory condition and completed fewer chains for food reinforcement as compared to the baseline session. However, during the second phase of the formalin test, animals showed decreased nociceptive behavior compared to formalin-injected animals that were not trained in the operant conditioning paradigm. During this phase, the trained animals exhibited maximal responsiveness for food reinforcement. These results demonstrate that the engagement of behaviors reflecting motivational drives to restore homeostasis depends on the intensity or degree of imbalance of the competing drives. More specifically, animals are motivated to attend to one state of imbalance at a time. PMID- 15531084 TI - Reduced brain edema and matrix metalloproteinase (MMP) expression by pre reperfusion infusion into ischemic territory in rat. AB - The aim in this study was to investigate whether our experimental model for stroke therapy, flushing the ischemic territory with saline prior to reperfusion, could ameliorate disruption of microvascular integrity by reducing matrix metalloproteinase (MMP) expression during reperfusion. Stroke in Sprague Dawley rats (n = 42) was induced by a 2-h right middle cerebral artery (MCA) occlusion using a novel intraluminal hollow filament. Prior to reperfusion, 24 of the ischemic rats received 6ml isotonic saline at 37 degrees C infused into the ischemic area through the filament. Brain edema was determined by comparing the percentage difference in brain volume between the right and left (contralateral to stroke site) hemispheres, while the expressions of MMP-2 and -9 mRNA were analyzed by real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR). A significant (p < 0.01) brain edema, determined by an increased brain volume of 19 +/- 4%, and overexpression of the mRNA encoding MMPs, determined by increased relative mRNA level ratio, were found in ischemic rats. The brain damage, in terms of brain edema (4 +/- 1%) and overexpression of MMPs, was significantly (p < 0.05) ameliorated as a result of saline flushing into the ischemic territory prior to reperfusion. This study has enhanced our understanding of the causal mechanisms by which the neuroprotective effect of ischemic area "flushing" can be achieved. PMID- 15531085 TI - Tetramethylpyrazine reduces ischemic brain injury in rats. AB - Tetramethylpyrazine (TMP), which is widely used in the treatment of ischemic stroke by Chinese herbalists, is one of the most important active ingredients of the traditional Chinese herbal medicine, Ligusticum wallichii Franchat (Chung Xiong). However, the mechanism by which TMP protects the brain is still not clear. We examined neuroprotective effects of TMP after transient focal cerebral ischemia using common carotid artery and middle cerebral artery occlusion model in rats and evaluated the involvement of anti-inflammation. TMP administrated intraperitoneally significantly protected the brain against ischemic insult as evidenced by the reduction in infarction volume, preservation of neurons, and decrease in brain edema. TMP markedly reduced cerebral ischemia/reperfusion induced inflammatory cell activation and proinflammatory mediator production. Moreover, TMP suppressed lipopolysaccharide/interferon-gamma-induced inflammation and prostaglandin E(2) production in cultured glial cells. Our findings suggest that one of neuroprotective effects of TMP against ischemic brain injury might involve its anti-inflammatory potential. PMID- 15531086 TI - TRAIL-related death receptors in normal, Lurcher and weaver mutant mouse brain. AB - In this study, we searched for murine analogues of the four death-receptor types (TRAIL-R1 to R4), targeted by the tumour necrosis factor related apoptosis inducing ligand (TRAIL), which were recently identified in the human brain. The expression of TRAIL-receptors in the normal murine brain was investigated using antibodies directed against different epitopes of the human TRAIL-receptors. Mouse mutants, in particular weaver and Lurcher with their well defined spatio temporal patterns of neurodegeneration in the cerebellum, the inferior olive and the substantia nigra, were used as a model for investigating a potential contribution of TRAIL-receptors to the genetically determined cell death observed in these mutants. Although all antibodies used, recognized the respective human antigens, only the murine analogue of the human TRAIL-R2 epitope was also identified in the mouse brain. Antisera against human TRAIL-R1, TRAIL-R3 and TRAIL-R4 failed to reveal any other murine TRAIL-receptor analogue. In normal mice, TRAIL-R2 is not universally expressed throughout the brain but rather restricted to specific neuronal populations predominantly consisting of large neurons. In weaver, the spatial patterns and relative densities of TRAIL-R2 labelling were virtually identical to those seen in wild-types during the period of cell death in the cerebellum and the substantia nigra. In Lurcher, TRAIL-R2 expression in cerebellar granule cells and inferior olivary neurons was identical to that in wildtypes but significantly reduced in Purkinje cells undergoing degeneration. Thus, although TRAIL-R2 is found to be expressed in various cell types of the murine brain, cell death in weaver and Lurcher mutants is apparently not accompanied by an upregulation of TRAIL-receptors. PMID- 15531087 TI - Communicating hands: ERPs elicited by meaningful symbolic hand postures. AB - Meaningful and meaningless hand postures were presented to subjects who had to carry out a semantic discrimination task while electrical brain responses were recorded. Both meaningful and control sets of hand postures were matched as closely as possible. The ERPs elicited by meaningless hand postures showed an anteriorly distributed N300 and a centro-posteriorly distributed N400 component. The N300 probably reflects picture-specific processes, whereas the N400-effect probably reflects processing in an amodal semantic network. The scalp distribution of the N400-effect, which is more posterior than usually reported in picture processing, suggests that the semantic representations of the concepts expressed by meaningful hand postures have similar properties to those of abstract words. PMID- 15531088 TI - Activation of extracellular signal-regulated kinases in the sciatic nerves of rats with experimental autoimmune neuritis. AB - To investigate whether the phosphorylation of extracellular signal-regulated kinase (ERK) is involved in autoimmune injury of the peripheral nervous system (PNS), the expression of phosphorylated ERK (p-ERK) was analyzed in experimental autoimmune neuritis (EAN) in rats. Western blot analysis showed that the level of p-ERK was increased significantly in the sciatic nerves of rats on days 14 (p<0.05) and 24 (p<0.01) post-immunization, compared with controls, and its reaction declined at day 30 post-immunization. Immunohistochemistry showed that p ERK protein was weakly expressed in Schwann cells and vascular endothelial cells in the sciatic nerves of CFA-immunized control rats. In EAN-affected sciatic nerves, p-ERK immunoreactivity was found mainly in ED1-positive macrophages on days 14 and 24 post-immunization. Moreover, on days 24 and 30 post-immunization, p-ERK immunoreactivity increased gradually in the Schwann cells of rat sciatic nerves with EAN. Based on these results, we postulated that the phosphorylation of ERK has an important role in the differentiation and survival of cells, including inflammatory cells and Schwann cells, in the rat sciatic nerve in EAN. Specifically, the activation of ERK in the recovery phase of EAN paralysis seems to be related in the survival of Schwann cells. PMID- 15531089 TI - Extending or projecting peripersonal space with tools? Multisensory interactions highlight only the distal and proximal ends of tools. AB - The effects of tool-use on the brain's representation of the body and of the space surrounding the body ('peripersonal space') has recently been studied within a number of disciplines in cognitive neuroscience, and is also of great interest to philosophers and behavioural ecologists. To date, most experimental findings suggest that tool-use extends the boundary of peripersonal space-visual stimuli presented at the tips of tools interact more with simultaneous tactile stimuli presented at the hands than visual stimuli presented at the same distance, but not associated with the tools. We studied the proposed extension of peripersonal space by tool-use by measuring the effects of three different tool use tasks on the integration of visual and tactile stimuli at three distances from participants' hands along two hand-held tools. When the tool-use task required using the shafts or the tips of the tools, visuotactile interactions were stronger at the tips of the tools than in the middle of the shaft. When the handles of the tools were used, however, visuotactile interactions were strongest near the hands and decreased with distance along the tools. These results suggest that tools do not simply 'extend' peripersonal space, but that just the tips of tools actively manipulated in extrapersonal space are incorporated into the brain's visuotactile representations of the body and of peripersonal space. PMID- 15531090 TI - Early maternal deprivation induces alterations in brain-derived neurotrophic factor expression in the developing rat hippocampus. AB - The effects of maternal deprivation (MD) during early postnatal life on the brain derived neurotrophic factor (BDNF) level were investigated in the present study. Wistar rats were assigned to either maternal deprivation or mother-reared control (MRC) groups. MD manipulation was achieved by separating rat pups from their mothers for 3h a day during postnatal days (PND) 10-15. At 16, 20, 30, and 60 days of age, the level of BDNF mRNA in the hippocampal formation of each group was determined using real-time PCR analysis. Early postnatal maternal deprivation of rat pups resulted in a significant increase in body weight at 60 days of age. The expression of BDNF mRNA in the hippocampus was significantly decreased at 16 days of age, and increased at 30 and 60 days of age. These data indicate that even a brief period of maternal deprivation during early postnatal life can affect hippocampal BDNF expression. PMID- 15531091 TI - Modulations of 'late' event-related brain potentials in humans by dynamic audiovisual speech stimuli. AB - Lipreading reliably improve speech perception during face-to-face conversation. Within the range of good dubbing, however, adults tolerate some audiovisual (AV) discrepancies and lipreading, then, can give rise to confusion. We used event related brain potentials (ERPs) to study the perceptual strategies governing the intermodal processing of dynamic and bimodal speech stimuli, either congruently dubbed or not. Electrophysiological analyses revealed that non-coherent audiovisual dubbings modulated in amplitude an endogenous ERP component, the N300, we compared to a 'N400-like effect' reflecting the difficulty to integrate these conflicting pieces of information. This result adds further support for the existence of a cerebral system underlying 'integrative processes' lato sensu. Further studies should take advantage of this 'N400-like effect' with AV speech stimuli to open new perspectives in the domain of psycholinguistics. PMID- 15531092 TI - A subcellular distribution of estrogen receptor-alpha is changed during artificially induced senescence of PC12 pheochromocytoma cells. AB - Although estrogen has been considered as a sex hormone for decades, recent reports suggest that estrogen might modulate the development and physiological function of the brain. In addition, the subcellular localization of estrogen receptors (ERs) has shown their presence within both the perinuclear cytoplasm and nuclei, suggesting that these ERs may differ functionally. We, therefore, assayed changes in the subcellular localization of ER-alpha immunoreactivity (IR) in rat pheochromocytoma PC12 cells during the artificial senescence induced by the telomerase inhibitor, 3'-azido-3'-deoxythymidine (AZT). After 2 months of culture with AZT, PC12 cells showed morphological and biochemical characteristics of cellular senescence. In the cells showing artificial senescence, the ER-alpha IR was mainly localized within the cytoplasm, whereas in control cells, ER-alpha IR was found only in the nuclei. Since senescence was induced by AZT, which inhibits the action of telomerase whenever the cells divide, the change in subcellular distribution of ER-alpha IR may be correlated with the length of the telomere. PMID- 15531093 TI - (S)-AMPA inhibits electrically evoked calcitonin gene-related peptide (CGRP) release from the rat dorsal horn: reversal by cannabinoid receptor antagonist SR141716A. AB - Previous studies in the hippocampus and cerebellum demonstrate that depolarisation of postsynaptic neurones stimulates the rapid synthesis and release of an endocannabinoid that retrogradely interacts with pre-synaptic CB(1) to modulate neurotransmitter release. This study evaluated whether depolarisation of second order neurones in the dorsal horn of the spinal cord by the AMPA receptor agonist, (S)-AMPA, would modulate sensory neurotransmission via release of endocannabinoids. Using an isolated rat dorsal horn with dorsal root attached in vitro preparation the release of calcitonin gene-related peptide (CGRP) after electrical stimulation of the dorsal roots was measured. Superfusion of either WIN55,212-2 (1 microM) or (S)-AMPA (1 microM) significantly attenuated CGRP release in a CB(1)-dependent manner (SR141716A, 5 microM). This provides indirect pharmacological evidence for an AMPA-evoked release of endogenous cannabinoids inhibiting peptide release from primary afferent neurons. This study confirms that CGRP release from the dorsal horn is modulated via CB(1) activation. Furthermore a depolarising stimulus also modulates CGRP release, potentially via the release of endogenous cannabinoids. PMID- 15531094 TI - D1 but not D2 dopamine receptors or adrenoceptors mediate dopamine-induced potentiation of N-methyl-d-aspartate currents in the rat prefrontal cortex. AB - Dopamine-glutamate interactions in the prefrontal cortex (PFC) are associated with higher order cognitive functions, and are involved in the pathophysiology of schizophrenia and addiction. Recordings with intracellular sharp microelectrodes and patch-clamp pipettes were used to investigate these interactions in layer V pyramidal cells of brain slices obtained from the rat PFC. Dopamine (100 microM) potentiated N-methyl-d-aspartate (NMDA; 10mM)-evoked depolarizations, but did not change those elicited by alpha-amino-3-hydroxy-5-methyl-4-isoxazole-4-propionic acid (AMPA; 1mM). Dopamine (100 microM) increased the amplitude of the NMDA (30 microM)-induced currents as well, and 1-phenyl-2,3,4,5-tetrahydro-(1H)-3 benzazepine-7,8-diol (SKF 38393; 1, 10 microM), a D(1) receptor agonist, concentration-dependently reproduced this effect. Furthermore, 7-chloro-8-hydroxy 3-methyl-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzapine hydrochloride (SCH 23390; 10 microM), a D(1) receptor antagonist, reversed both the dopamine- and the SKF 38393-evoked potentiation. The D(2) receptor agonists lisuride and quinpirole (10 microM both), as well as noradrenaline (100 microM) failed to mimic the stimulatory effect of dopamine. Isoproterenol (1, 10 microM) concentration dependently facilitated NMDA responses. However, neither this effect at 10 microM nor that of dopamine at 100 microM could be antagonized by propranolol (10 microM), a non-selective beta adrenoceptor blocker. The isoproterenol-induced facilitation of NMDA currents was abolished by SCH 23390 (10 microM). The results indicate that dopamine potentiates NMDA responses in layer V pyramidal cells of the PFC solely by activating D(1) receptors. D(2) receptors and alpha or beta adrenoceptors are not involved in the dopamine-NMDA interaction. PMID- 15531095 TI - Relationship between suicidal behavior of psychotic inpatients and serotonin transporter gene in Han Chinese. AB - Strong evidence indicates that central serotoninergic system dysfunction is associated with suicidal behavior. The relationship between the serotonin transporter protein (5-HTT) gene of psychotic patients in Han Chinese with suicidal behavior was studied since the 5-HTT plays an important role in serotoninergic transmission. Two hundred and seventy-two psychotic inpatients with suicidal behavior (suicide group) and 275 psychotic inpatients without suicidal behavior (patient control group) were recruited from six mental health facilities in Shanghai, China. Another 628 healthy adults (healthy control group) without suicidal behavior and mental disorder history were enrolled from Shanghai, China. All subjects were Han Chinese. The genotypes and the alleles of 5-HTT genes of the three groups were examined by PCR amplification. Neither the intron 2 variable number tandem repeat (VNTR) polymorphism and 5-HTTLPR (5-HTT gene-linked polymorphic region), nor the haplotype frequencies of this gene have significant differences between the suicide group and the two control groups. However, there was a significant difference in 5-HTTLPR polymorphism between inpatients with a first episode of suicidal behavior and those with recurrent suicidal behavior. Except for the possible heterogeneity between inpatients with a first episode of suicidal behavior and those with more than one, the 5-HTT gene was unlikely to be associated with suicidal behavior of psychotic patient in Han Chinese. PMID- 15531096 TI - Effects of venlafaxine on p90Rsk activity in rat C6-gliomas and brain. AB - The intracellular actions of the antidepressant, venlafaxine, were studied in C6 gliomas using a phosphoproteomics approach. Long-term pre-treatment of C6-gliomas with venlafaxine followed by an acute challenge with isoproterenol (a beta adrenoceptor agonist), resulted in increased p90Rsk phosphorylation (three-fold) versus control levels (isoproterenol alone). The effect of venlafaxine pre treatment on p90Rsk activity was dose-dependent (EC(50)=3.75nM) in C6 gliomas. In rat brain sections, intense immunoreactive phospho-p90Rsk labeling was observed for both neurons and glia, especially in cortical layers II/III and hippocampal formations. In vivo studies demonstrated an intense but similar distribution pattern of phospho-p90Rsk staining after chronic venlafaxine dosing of rats compared to naives and no region-specific drug effect was observed in vivo. In conclusion, our findings suggest that some of the centrally-mediated benefits of venlafaxine in depression may be due to its intracellular properties especially on the neuro-glial circuitry and MAPK/p90Rsk-dependent pathways at an early stage. PMID- 15531097 TI - Distribution and synaptic localization of nicotinic acetylcholine receptors containing a novel alpha7 subunit isoform in embryonic rat cortical neurons. AB - Neuronal nicotinic acetylcholine receptors (nAChRs) containing the alpha7 subunit isoform, alpha7-2 (alpha7-2-nAChRs), have previously been found to form functional homopentameric channels that desensitize slowly and bind alpha bungarotoxin (alphaBgt) in a rapidly reversible manner. This isoform incorporates a novel cassette exon in the extracellular, ligand binding domain of the native receptor. Although this alpha7 subunit isoform has been detected in peripheral ganglia as well as in the central nervous system, little is known about the cellular function of alpha7-2-nAChRs. Co-localization immunocytochemical studies were conducted in an embryonic rat cultured cortical neuron model using a polyclonal antibody (Ab 87) raised against the amino acid sequence of the cassette exon, in combination with (1) an antibody that recognizes all known alpha7-nAChRs, (2) alphaBgt, and (3) antibodies directed against multiple cellular markers. The pattern of alpha7-2-nAChR expression was consistent with alpha7 staining in general, based on co-distribution of mAb319 and alphaBgt signals. However, alpha7-2-nAChRs clearly represent a distinct subset of alpha7 receptors. The alpha7-2-nAChR subtype was found throughout the cell-soma surface and was localized to a subpopulation of dendrites. Punctate staining characteristic of synaptic alpha7-2 targeting was observed at post-synaptic densities and intermittently at pre-synaptic locations. The alpha7-2 subunit was expressed on both GABAergic and non-GABAergic neurons. These studies reveal that receptors containing the alpha7-2 subunit constitute a subpopulation of alpha7 nAChRs and likely participate in cell-to-cell signaling in developing synapses of central neurons. PMID- 15531098 TI - Effects of pituitary adenylate cyclase activating polypeptide in retinal degeneration induced by monosodium-glutamate. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a pleiotropic neuropeptide with a wide range of effects in the central and peripheral nervous systems. PACAP has well-documented neurotrophic and neuroprotective actions in both in vitro and in vivo models of different neuronal injuries. The aim of the present study was to investigate the possible neuroprotective effect of PACAP in retinal degeneration induced by monosodium-glutamate (MSG) in neonatal rats. Preceding the MSG treatment, PACAP (1 or 100pmol/5mul) was injected unilaterally into the vitreous body on postnatal days 1, 5 and 9. Immediately after the PACAP treatment, pups were treated with 2mg/g body weight MSG subcutaneously. At 3 weeks of age, rats were sacrificed and retinas were removed and processed for histological examination. Our results show that MSG treatment caused severe degeneration, primarily of the inner retinal layers. The thickness of the entire retina was only approximately half of that of the normal retinas, and the inner nuclear layer seemed to be fused with the ganglionic cell layer, with no discernible inner plexiform layer. Retinas of animals treated with 1pmol PACAP showed a similar degree of degeneration. However, retinas of rats treated with 100pmol PACAP showed significantly less damage, with clearly distinguishable inner retinal layers. In summary, our present study shows that local PACAP treatment could attenuate the retinal degeneration induced by the excitotoxic effects of glutamate. PMID- 15531099 TI - Effect of hypoxia on protein tyrosine kinase activity in cortical membranes of newborn piglets--the role of nitric oxide. AB - Previous studies have shown that cerebral hypoxia results in increased tyrosine phosphorylation of cerebral cortical cell membrane proteins as well as nuclear membrane anti-apoptotic protein, Bcl-2. The present study tests the hypothesis that hypoxia results in increased protein tyrosine kinase activity in cortical cell membranes of newborn piglets and that the inhibition of neuronal NOS by administration of 7-nitroindazole sodium salt (7-NINA), a selective inhibitor of nitric oxide synthase (NOS), will prevent the hypoxia-induced increase in protein tyrosine kinase activity. To test this hypothesis, protein tyrosine kinase activity was determined in cerebral cortical membranes of 2- to 4-day-old newborn piglets divided into normoxic (n=6), hypoxic (n=5) and 7-NINA-treated hypoxic (n=5) (7-NINA, 1mg/kg, i.p., prior to hypoxia) groups. Tissue hypoxia was achieved by exposing the animals to an FiO(2) of 0.07 for 60 min and was documented biochemically by determining tissue ATP and phosphocreatine (PCr) levels. Cortical P(2) membranes were isolated and protein tyrosine kinase activity determined by (33)P incorporation into a specific peptide substrate for 15 min at 37 degrees C in a medium containing 100 mM HEPES, pH 7.0, 1mM EDTA, 125 mM MgCl(2), 25 mM MnCl(2), 2mM DTT, 0.2 mM sodium orthovanadate, 2mM EGTA, 150 microM tyrosine kinase peptide substrate [Lys 19] cdc2(6-20)-NH(2), (33)P-ATP, and 10 microg of membrane protein. Protein tyrosine kinase activity was determined by the difference between (33)P incorporation in the presence and absence of specific peptide substrate and expressed as pmol/mg protein/h. The ATP values in the normoxic, hypoxic and 7-NINA-treated hypoxic animals were ATP: 4.57+/-0.45 micromol/g, 1.29+/-0.23 micromol/g (p<0.05 versus normoxic) and 1.50+/-0.14 micromol/g brain (p<0.05 versus normoxic), respectively. The PCr values in the normoxic, hypoxic and 7-NINA-treated hypoxic animals were: 3.77+/ 0.36 micromol/g, 0.77+/-0.13 micromol/g (p<0.05 versus normoxic) and 1.02+/-0.24 micromol/g brain (p<0.05 versus normoxic), respectively. Protein tyrosine kinase activity in the normoxic, hypoxic and the 7-NINA-treated groups was 378+/-77 pmol/mg protein/h, 854+/-169 pmol/mg protein/h (p<0.05 versus normoxic) and 464+/ 129 pmol/mg protein/h (p<0.05 versus hypoxic), respectively. The data show that cerebral tissue hypoxia results in increased protein tyrosin kinase activity in cortical membranes of newborn piglets and pretreatment with 7-NINA prevents the hypoxia-induced increase in protein tyrosine kinase activity. We conclude that the hypoxia-induced increase in protein tyrosine kinase activity is NO-mediated. We propose that the hypoxia-induced increase in protein tyrosine kinase activity leading to increased phosphorylation of Bcl-2 is a critical link to hypoxic neuronal injury pathway. PMID- 15531100 TI - Using virtual reality to analyze links between handball thrower kinematics and goalkeeper's reactions. AB - This work investigates the design of a new method to evaluate the importance of visual elements taken into account by a handball goalkeeper facing a thrower. Virtual reality was used to design and reproduce standardised situations in a controlled environment. Under such conditions, it was possible to isolate for investigation one visual element in the thrower's gestures. The goalkeeper's movements were recorded in order to compare his reactions to two separate throws where only one visual element was modified. Our systems allowed us to measure and record the effects of small changes in the thrower's movements. With the numerical values we obtained from our results we were able to come up with a scale of significance for each isolated element. These preliminary results look promising for neuroscience, allowing us to better understand the strategies used in duel situations. PMID- 15531101 TI - Increased WDR spontaneous activity and receptive field size in rats following a neuropathic or inflammatory injury: implications for mechanical sensitivity. AB - Spontaneous activity and receptive field size for spinal wide dynamic range (WDR) neurons were measured and related to the mechanical allodynia in both neuropathic (L5-L6 ligation, 14 days post-injury) and complete Freund's adjuvant-inflamed rats (CFA, 2 days post-injury). The size of the WDR receptive field located on the hindpaw expanded significantly (p<0.01) following both modes of injury, with no difference between CFA and neuropathic animals. Likewise, the spontaneous firing of WDR neurons was significantly elevated following both the CFA (4.4+/ 0.6 spikes/s, p<0.01) and neuropathic (3.2+/-0.3 spikes/s, p<0.05) injuries compared to naive (2.1+/-0.2 spikes/s) and sham-neuropathic (1.9+/-0.3 spikes/s) rats. Furthermore, the spontaneous WDR activity recorded from CFA rats was also significantly greater (p<0.05) than neuropathic rats. Mechanical allodynia, as measured by application of a von Frey hair stimulus, was observed from both CFA and neuropathic rats, however, the degree of sensitivity was significantly greater (p<0.01) for the CFA animals. These data suggest that the differences in mechanical sensitivity between CFA and neuropathic rats may be related to their respective changes in WDR spontaneous activity, but not to the changes in receptive field size, and is further demonstration of the importance of spontaneous WDR activity in determining mechanical sensitivity following injury. PMID- 15531102 TI - Time dependent alterations in dopamine tissue levels and metabolism after experimental traumatic brain injury in rats. AB - Several studies have demonstrated alterations in the dopamine (DA) system after traumatic brain injury (TBI). The present study investigated the temporal changes in DA tissue levels and metabolism at 1-h or 1, 7, 14, and 28 days after cortical impact or sham injury in rats. DA and DOPAC levels were measured by HPLC in the frontal cortex (FC) and striatum. DA levels were significantly increased at 1h in the contralateral FC and at 1 day in the ipsilateral FC versus respective sham groups. DA and DOPAC levels were significantly increased bilaterally at 1h in the striatum versus sham. These data indicate that TBI induces an early increase in DA and DOPAC, which returns to sham levels over time. PMID- 15531103 TI - Activation of metabotropic glutamate receptors does not alter the phosphorylation state of GluR1 AMPA receptor subunit at serine 845 in perirhinal cortical neurons. AB - Activation of group I and group II metabotropic glutamate receptors (mGluRs) is thought to be required for long-term depression (LTD) of alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionate (AMPA) receptor-mediated synaptic transmission in the perirhinal cortex. However, little is known about how activation of mGluRs leads to this form of synaptic plasticity. AMPA receptor phosphorylation has been implicated in several forms of modulation of synaptic transmission. In the CA1 area of the hippocampus, N-methyl-d-aspartate (NMDA) receptor-dependent LTD is associated with the reduced phosphorylation of the GluR1 AMPA receptor subunit at serine 845 (GluR1-S845). Immunoblot analysis of perirhinal cortical neurons using GluR1 and GluR1-S845 phosphorylation state specific antibodies showed that stimulation of adenylyl cyclase (AC) with forskolin (FSK) dramatically increased PKA-mediated phosphorylation of GluR1-S845. However, selective or simultaneous application of mGluR5 agonist (S)-3,5-dihydroxyphenylglycine (CHPG) and mGluR2/3 agonist (2S,2'R,3'R)-2-(2',3'-dicarboxycyclopropyl)glycine (DCG IV) did not produce detectable changes in GluR1-S845 phosphorylation. These results indicate that in the perirhinal cortex mGluR activation does not alter the phosphorylation state of GluR1-S845. Therefore, it is likely that the process involved in the modification of AMPA receptors in mGluR activation dependent LTD in the perirhinal cortex is mechanistically distinct from NMDA receptor-mediated LTD described in hippocampal neurons. PMID- 15531104 TI - Upregulation of osteopontin in Schwann cells of the sciatic nerves of Lewis rats with experimental autoimmune neuritis. AB - We examined the expression of osteopontin (OPN) in the sciatic nerves of rats with experimental autoimmune neuritis (EAN), using immunohistochemistry and immunoblotting, to study its involvement in the pathogenesis of autoimmune peripheral nervous system diseases. Constitutive OPN expression was detected in some Schwann cells; expression was increased after immunization with adjuvant alone. At day 14 after induction of EAN, many Schwann cells had a granular pattern of immunoreactivity, whereas very few inflammatory cells were OPN positive. Even after recovery from hindlimb paralysis, at 24 days post immunization, OPN expression remained elevated in the Schwann cells. The results suggest that OPN expression in Schwann cells is easily induced by immunostimulation, and further enhanced by the inflammatory reaction in EAN. Continued elevation of OPN after recovery may represent a functional recovery after a transient inflammatory insult. PMID- 15531105 TI - Supraclinical concentrations of dexmedetomidine evoke norepinephrine release from rat cerebrocortical slices possible involvement of the orexin-1 receptor. AB - Dexmedetomidine is a highly selective alpha(2)-agonist and reduces norepinephrine release from several neuronal tissues. However, supraclinical concentrations of dexmedetomidine have been reported to increase norepinephrine release from cardiac stores. In addition, some report using microdialysis shows that intrathecal clonidine increased norepinephrine release from the dorsal horn in mid-thoracic spinal cord but dexmedetomidine did not. Thus, in the present study we have studied effects of dexmedetomidine on norepinephrine release from rat cerebrocortical slices and compared this with clonidine. We have also used a selective alpha(2)-antagonist yohimbine and an orexin-1 receptor antagonist SB 334867 to examine whether the effects of dexmedetomidine on norepinephrine release are mediated via alpha(2)-adrenergic or orexin (OX) receptors. In addition, concentrations of orexin A in the evoked sample were also measured. Dexmedetomidine significantly increased norepinephrine release (basal=100%) from rat cerebrocortical slices in a concentration-dependent manner with E(max) 377.3+/-8.6% and pEC(50) 6.12+/-0.07, whereas clonidine significantly reduced the release with E(max) 62.1+/-6.8% and pEC(50) 4.55+/-0.25. Yohimbine (10(-5)M) did not affect the concentration-response curve of dexmedetomidine for norepinephrine release. However, SB-334867 concentration-dependently antagonized dexmedetomidine evoked norepinephrine release with I(max) 91.0+/-9.4% and pIC(50) 5.99+/-0.18. Orexin A concentrations did not differ between the samples. Thus, supraclinical concentrations of dexmedetomidine increase norepinephrine release from rat cerebrocortical slices, and this release may be mediated via OX(1) but not alpha(2)-adrenoceptors. PMID- 15531106 TI - Upregulation of cellular prion protein (PrPc) after focal cerebral ischemia and influence of lesion severity. AB - The pathological isoform of the prion protein (PrP(Sc)) has been identified to mediate transmissible spongiform encephalopathies like Creutzfeldt-Jakob disease (CJD). In contrast, the physiological function of the normal cellular prion protein (PrP(c)) is not yet understood. Recent findings suggest that PrP(c) may have neuroprotective properties and that its absence increases susceptibility to oxidative stress and neuronal injury. To determine whether PrP(c) is part of the cellular response to neuronal injury in vivo, we investigated PrP(c) regulation after severe and mild focal ischemic brain injury in mice using the thread occlusion stroke model. Western Blot and ELISA analysis showed a significant upregulation of PrP(c) in the ischemic hemisphere at 4 and 8h after onset of permanent focal ischemia, which was no longer detectable at 24h after lesion induction when compared to control animals. In contrast, transient focal ischemia (60 min) did only lead to slightly but not significantly elevated PrP(c) levels in the ischemic hemisphere when compared to controls. These results demonstrate that cerebral PrP(c) is upregulated early in response to focal cerebral ischemia. The extent of upregulation, however, seems to depend on the severity of ischemia and may therefore reflect the extent of ischemia induced neuronal damage. Given the known neuroprotective effects of PrP(c) in vitro, ischemia-induced upregulation of cerebral PrP(c) supports the hypothesis that, as part of an early adaptive cellular response to ischemic brain injury, PrP(c) may be involved in the regulation of ischemia-induced neuronal cell death in vivo. PMID- 15531107 TI - Unusual functional properties of homo- and heteromultimeric histamine-gated chloride channels of Drosophila melanogaster: spontaneous currents and dual gating by GABA and histamine. AB - Histamine acts as a neurotransmitter of photoreceptors in insects and other arthropods, where it directly activates a chloride channel and mediates rapid inhibitory responses. Homo- and heteromultimeric histamine-gated ion channels formed by HisCl-alpha2 or HisCl-alpha1 + alpha2 subunits from Drosophila melanogaster were characterized by two-electrode voltage-clamp measurements of functionally expressed ion channels in Xenopus laevis oocytes. The sensitivity of heteromultimeric histamine receptors with an EC(50) of 2.3 microM is lower than that of either homomultimeric receptor. They can be further distinguished from the homomultimeric channels by their reduced sensitivity to d-tubocurarine. Heteromultimeric channels generate a spontaneous current in the absence of any agonist. This spontaneous current can be blocked in the absence of an agonist by d-tubocurarine and the histamine antagonists cimetidine, thioperamide and pyrilamine. Homomultimeric HisCl-alpha2 channels are dually gated by histamine (IC(50)=9.4 microM) and GABA (IC(50)=1.0mM), both of which are full agonists. The action of both agonists can be blocked with comparable IC(50) values by the histamine antagonists cimetidine, thioperamide and pyrilamine but not by the GABA antagonist bicuculline. Picrotoxin blocked with an IC(50) of 403 microM. Our data show that histamine and GABA act on the same ion channel, which thus might function as a site of integration of the action of different neurotransmitters. PMID- 15531108 TI - Haloperidol and clozapine, but not olanzapine, induces oxidative stress in rat brain. AB - Typical and atypical antipsychotic drugs have been shown to have different clinical and behavioral profiles. Haloperidol (HAL) is a typical neuroleptic that acts primarily as a D2 dopamine receptor antagonist. It has been proposed that reactive oxygen species play a causative role in neurotoxic effects induced by HAL. We evaluated oxidative damage in rat brain induced by chronic HAL, clozapine (CLO) or olanzapine (OLZ) administration. Adult male Wistar rats received daily injections of Hal (1.5mg/kg), CLO (25mg/kg) or OLZ (2.5, 5.0 or 10.0mg/kg). Control animals were given saline (SAL; NaCl 0.9%). Thiobarbituric acid reactive substances (TBARS) and protein carbonylation were measured in the hippocampus (HP), striatum (ST) and cortex (CX). TBARS was increased in the striatum after HAL treatment. In contrast, there was a decrease of TBARS levels induced by HAL, CLO and OLZ treatments in the cortex. Protein carbonyls after HAL and CLO treatment was increased in the hippocampus, compared to control. In hippocampus, OLZ did not show significant difference to control in both oxidative parameters. Our findings demonstrated that atypical antipsychotic CLO produced less oxidative damage than HAL and we did not find oxidative damage induced by OLZ. PMID- 15531109 TI - The modulation of the Ne-like wave on correct responses foreshadows errors. AB - In reaction time (RT) tasks, event-related potentials (ERPs) reveal a response locked negative wave when subjects commit errors. This wave, termed "error negativity" (Ne) or "error-related negativity" (ERN), is thought to index response-monitoring processes. With conventional monopolar recordings, this negativity is hardly seen on correct responses, likely overlapped by a large positive wave. Indeed, after Laplacian transformation (a spatial high-pass filter), a small Ne-like wave is unmasked. Recently, it has been shown that the positivity on monopolar recordings was larger for correct responses preceding an error than for correct responses preceding a correct trial. After Laplacian transformation, it appears that this effect is due, at least in part, to a decrease of the Ne-like wave on correct responses preceding an error. This result indicates that, as the Ne on errors, the Ne-like wave on correct responses is sensitive to performance and hence is likely related to response-monitoring processes. PMID- 15531110 TI - Aglycemia and ischemia depress monosynaptic and polysynaptic reflexes in neonatal rat spinal cord in vitro by involving different types of 5-hydroxytryptamine receptors. AB - Involvement of 5-hydroxytryptaminergic (5-HT) system for aglycemia- and ischemia induced depression of the spinal reflexes was evaluated. The monosynaptic (MSR) and polysynaptic (PSR) reflex potentials were elicited in the ventral root by stimulating the corresponding dorsal root in an isolated spinal cord from neonatal rat. Superfusion of aglycemic (glucose-free) or ischemic (glucose- and O(2)-free) solution produced a time-dependent depression of the spinal reflexes and abolished them within 35 min. The time required to produce 50% depression of the reflexes (T-50) was around 19 and 13 min for aglycemia and ischemia, respectively. Spiperone (5-HT(2A) antagonist) and ketanserin, (5-HT(2A/2C) antagonist) blocked the aglycemia-induced depression of the reflexes completely while ondansetron (5-HT(3) antagonist) attenuated it partially (as abolition times were around 50 min). Ischemia-induced depression was blocked up to 50 min by ketanserin or ondansetron but not by spiperone. In the presence of ketanserin or ondansetron, the reflexes were abolished by 60 min while in spiperone the reflexes were abolished by 30 min. In 5,7-dihydroxytryptamine treated rats, aglycemia depressed the reflexes by 45 min (greater than control, P<0.05) while the time for ischemia-induced depression was not different from the control response. The results indicate that aglycemia involves mainly 5-HT(2) receptors while ischemia involves 5-HT(3) receptors. PMID- 15531111 TI - The NR1 subunit of the glutamate/NMDA receptor in the superior temporal cortex in schizophrenia and affective disorders. AB - The NMDA receptor has been implicated in the pathophysiology of several diseases including schizophrenia and affective disorders. We have investigated the NR1 subunit of the NMDA receptor in a well-defined series of psychiatric cases using radioligand binding and quantitative immunoblotting techniques. Saturable radioligand binding of [(3)H]L-689,560 to the glycine site on this subunit of the NMDA receptor was undertaken in superior temporal cortex of patients with schizophrenia, bipolar disorder, depression and matched control subjects. A tendency towards an increased receptor density was found in schizophrenia. A significant decrease in NMDA receptor density below control value was found in both bipolar and depressive disorders. The immunoblotting technique was used to identify NR1 protein in the same series of cases of which two bands were identified consistent with NR1 splice variants. A tendency to a decrease in the density of the NR1 upper band below control values was found in bipolar and depressed patients, but not schizophrenics. Consistent with this observation, the ratio between the upper and lower NR1-immunoreactive bands showed a significant decrease in bipolar disorder, although the ratio in depression did not reach significance. No significant difference was found in the NR1 lower band in any patient group compared with control. The finding of an increase NMDA receptor density in schizophrenia is consistent with the previous reports, with a possible compensatory response to glutamatergic deficits in superior temporal cortex in schizophrenia. The findings in affective disorders are interesting in respect of reports of cortical NMDA receptor deficits in suicide victims, although antidepressant drug treatment may contribute to these changes. PMID- 15531112 TI - [Detection of HBV DNA by sensitive techniques and definition of chronic VHB infection by pre-C-C mutants]. AB - Our purpose is to assess the question of the definition of hepatitis B virus pre C-C mutant-chronic infection, according to the level of the viral load at the era of very sensitive techniques of quantification of HBV DNA. PMID- 15531113 TI - [Mutation analysis of ISDR and V3 domains of hepatitis C virus NS5A region before interferon therapy with or without ribavirin]. AB - AIM OF THE STUDY: The hepatitis C virus (HCV) non-structural NS5A protein has been controversially implicated in the resistance of HCV to interferon therapy in clinical studies. In Japan, mutations in the interferon sensitivity-determining region (ISDR) in the NS5A gene were associated with response to interferon therapy in patients infected with genotype 1b. In contrast, studies from Europe did not confirm such association. More recently, it has been suggested that the V3 domain outside the putative ISDR might also have amino acids changes that may be involved in the resistance to IFN. In this study, the relationship between NS5A mutations in ISDR and V3 domains and virological response to therapy were investigated. MATERIALS AND METHODS: The NS5A gene was sequenced from 35 HCV genotype 1b infected patients at D0 of a prospective clinical trial of interferon therapy and interferon plus Ribavirin combination therapy. RESULTS: In the ISDR domain, we did not observe any significant differences in amino acids changes between responders (1.7 +/- 1.8, n = 19, range 0-6) and non-responders (1.1 +/- 0.8, n = 14, range: 0-3), (P = 0.483), to therapy before the beginning of treatment. In the V3 domain, we found more mutations in responders (6.5 +/- 1.9, range: 2-11) than in non-responders (4.7 +/- 1.2, range: 3-8) (P = 0.0013), before the beginning of treatment. CONCLUSION: Our results confirm that, in Europe, the ISDR domain is not predictive for treatment success but suggest that the V3 domain have greater variability in responders than non-responders. PMID- 15531114 TI - [Evaluation of two commercial enzyme immunoassays for diagnosis of hepatitis C in the conditions of a virology laboratory]. AB - BACKGROUND: Most studies which evaluate antibody detection assays are conducted on blood donors specimens, i.e healthy individuals. Sera collected in patients, vs healthy individuals, can make serological tests difficult because of possible non specific reactions interfering with serological tests. The aim of this work was to compare the specificity and the sensitivity of two commercial automated assays for the detection of hepatitis C virus antibody, Monolisa anti-HCV Plus on the Evolis automate (Biorad) and Axsym anti-HCV 3.0 (Abbott). PATIENTS AND METHOD: The prospective study of specificity included 2020 routine serum samples sent to our virology laboratory. The sensitivity was established with eight commercially available HCV seroconversion panels. RESULTS: The Monolisa and the Axsym assays showed a specificity of 99.64 and 99.12%, respectively. Of 49 specimens from eight commercially available HCV seroconversion panels, the number of positive results was 21 and 24 for the two tests, respectively. CONCLUSION: A statistical analysis of specificity and sensitivity results proved no significant difference between the two tests. Nevertheless, the Monolisa kits could be preferred for its more homogeneous sensitivity than the Axsym test and for its apparent better specificity. The final choice of a kit should also take into account the easiness to perform and an optimal integration in the usual practice of the concerned laboratory. PMID- 15531115 TI - [Immunoblot in the serological diagnosis of hepatitis C virus infection]. AB - The objective of the study was to assess three immunoblot assays, the Deciscan HCV Plus, the Riba and the Inno-Lia, on 44 discordant samples with three EIA kits. These immunoblots were considered as confirmation reagents. A result was considered as a false positive by anti-HCV antibody assay if the three immunoblots were negative or if two immunoblots were negative with the third being indeterminate and a negative virological genomic diagnosis observed on all the samples. The result was positive if at least two immunoblots out of three were positive. Thus, 34 samples were considered as false positive and ten samples were excluded because it was impossible to conclude between true or false positive result. The 44 discordant results were never confirmed as positive by the use immunoblot or PCR. The three immunoblots were negative for half of the samples and two immunoblots and one indeterminate were observed for 77% of the samples. The false positive results by the Monolisa assay were more often found indeterminate with the Deciscan assay than with the other immunoblots. That was also checked for Vitros/Riba pair. One of the explanations could be the use of common antigens for the reagents from the same manufacturer. The Inno-Lia test is the most specific immunoblot according to the results obtained in our study. PMID- 15531116 TI - [Comparison of hepatitis C viral RNA and core antigen kinetics in the therapeutic follow up of hepatitis C virus and human immunodeficiency virus co-infected patients, treated by bitherapy interferon-ribavirin, within the framework of RIBAVIC protocol]. AB - AIM OF STUDY: The RIBAVIC protocol, established by ANRS in 2001 and closed in 2003, compared the efficacy and the tolerance of two bitherapy anti-Hepatitis C Virus for HIV-HCV co-infected patients: IFN-ribavirin and PEG-IFN-ribavirin for 48 weeks. Two hundred patients from protocol were tested for hepatitis C virus core antigen, to study this viral marker kinetics, before and under treatment, in comparison with hepatitis C virus RNA evolution. MATERIAL AND METHODS: The available samples for the 204 patients of our study were tested for RNA detection (COBAS AMPLICOR v2.0, Roche Diagnostics) and quantification (VERSANT HCV RNA v3.0, Bayer Diagnostics) and for quantification of core antigen (Ortho trak-C Assay, Ortho Clinical Diagnostics). The viral kinetics were established from samples quantified at D0, W2, W4, W12, W24, W48, W52, W72 (W =week), according to virological response assessed by PCR, six month after the end of treatment (non responders, sustained responders, relapsers et breakthroughs). RESULTS: We obtained, for each type of response, similar evolution of both viral markers. Trak-C assay show to be enough sensitive, with similar results whatever genotype of hepatitis C virus. The Pearson's correlation is excellent (R =0.94; P <0.001). The intergenotype correlation is correct too, whatever HCV genotype (1, 2, 3, 4). CONCLUSIONS: The HCV core antigen quantification by trak-C assay is a new tool for the follow-up of the treatment of patients with chronic hepatitis C and HIV co-infected. PMID- 15531117 TI - [Risk factors for Abacavir-induced hypersensibility syndrome in the "real world"]. AB - Abacavir (ABC) is a generally well-tolerated NRTI. However, up to 5% of patients may develop hypersensitivity syndrome (HSS) within the first weeks of treatment. The objectives of this study were to describe the side effects of ABC, to evaluate the incidence of the ABC-HSS, and to identify the risk factors of HSS after first exposure to ABC in a cohort of patients followed up in a university HIV clinic. METHODS: The charts of all HIV-infected patients who started ABC between February 1998 and May 2002 were reviewed. HSS was defined as the onset, within 8 weeks of ABC initiation, of either a skin rash associated with at least one of the following symptoms (fever, gastrointestinal symptoms, respiratory symptoms, myalgia, malaise) or at least three of the above symptoms in the absence of rash. A multivariate logistic regression analysis was performed to identify risk factors of HSS. RESULTS: Of the 191 patients studied (134 M, 57 F, mean age 39 years), 53 (27.8%) presented with manifestations that were regarded as potential side-effects of ABC. Ten (5.2%) developed HSS, none of whom died. Two factors were independently associated with an increased risk of HSS: history of allergy to nevirapine (OR 8.1, 95% CI 1.6-40.5, p = 0.02), and being naive to ART (OR 5.8, 95% CI 1.2-28.5, p = 0.04). CONCLUSION: This study "in the real world" confirms that the incidence of ABC-induced HSS is of about 5%. It also confirms that HSS occurs more frequently in patients with a history of allergy to nevirapine and in ART-naive patients. PMID- 15531119 TI - [Assessment of the diagnostic value of RT-PCR on amniotic fluid for prenatal diagnosis of congenital rubella infection]. AB - AIM OF THE STUDY: To assess the diagnostic value of RT-PCR on amniotic fluid (AF) for prenatal diagnosis of congenital rubella infection. MATERIAL AND METHODS: RT PCR on AF was compared to specific IgM antibody detection in foetuses and/or newborns in 45 pregnant women with confirmed primary infection. RESULTS: specificity of RT-PCR was 100% and sensitivity ranged between 83 and 95%. CONCLUSION: RT PCR may be considered as a valuable tool for prenatal diagnosis of foetal rubella infection. PMID- 15531118 TI - [Genetic polymorphism of cytomegalovirus strains responsible of congenital infections]. AB - OBJECTIVES: Congenital Cytomegalovirus (CMV) infection is the main cause of neurological handicap in young children. The objective for studying genetic polymorphism of strains responsible for congenital infection is to identify CMV strains or groups of strains which would be more frequent in this context and/or which would be responsible for more severe congenital infection. METHODS: In this paper, we report and analyze the literature concerning the genetic polymorphism of CMV strains responsible of congenital infection, in the genes coding for the envelop protein B and the non structural UL144 protein and in the CMV short tandem repeats. RESULTS AND CONCLUSION: All UL144 and gB genotypes can be vertically transmitted from mothers to fetuses, none of these studies has shown any link between the genotypes and the severity of congenital disease. Moreover, no link between short tandem repeats polymorphism and severity of congenital disease has been demonstrated. However, short tandem repeats analysis may be a powerful tool to study the epidemiology of CMV congenital infections. PMID- 15531120 TI - [Usefulness of panfungal PCR for the diagnosis of fungal infection in immunocompromised patients]. AB - The diagnostic of invasive fungal infection is often difficult because of the low sensitivity of fungal culture from infected tissues. Here we have assessed the ability of a panfungal PCR targeted on the DNA region encoding the RNA genes followed by sequencing of the amplification products to detect and identify fungi from tissue biopsies. This assay allowed us to identify the microorganism responsible for an invasive fungal infection in three of our patients. In two cases, hepatosplenic candidiasis was suspected and Candida albicans DNA was detected from liver biopsies. The third patient was cared for a thymome and developed a manubrium osteitis caused by Scedosporium apiospermum. PMID- 15531121 TI - [Role of Abelcet in children with febrile neutropenia]. AB - AIM OF STUDY: In order to optimise the use of new forms of Amphotericine B (Ampho B), a decisional tree was created at the end of 2001 in the paediatric hemato oncology unit for the empirical antifungal treatment in febrile neutropenic children: the standard remained conventional Ampho B and Abelcet was proposed in case of antecedent or occurrence of a deterioration of the renal function (DRF). In order to validate the place of Abelcet we initiated a retrospective study over year 2002. RESULTS: 21 treatments were begun in 14 children for a median duration of 8 days (1-48 days). Three kind of indications were found: DRF antecedent (10 episodes: A group), DRF occurrence during a treatment with conventional Ampho B (7 episodes: B group), age lower than 1 year (3 episodes). 81% of the children were thus treated according to the decisional tree. The clinical tolerance was good in 90% of the cases, with a premedication in half of the cases. The study of the renal function showed a good renal tolerance for 6 episodes out of 9 evaluable in A group, 3 resolutions and 2 stabilisation of the renal failure for the 5 evaluable episodes of the B group. Seven to ten days of treatment by Abelcet were necessary to obtain the renal failure resolved. CONCLUSION: This study confirms the interest of Abelcet in the empirical antifungal treatment in febrile neutropenic children and specially in children having antecedents of DRF related or not to a treatment with conventional Ampho B. PMID- 15531122 TI - Postmenopausal testosterone therapy and breast cancer risk. AB - BACKGROUND: Testosterone therapy is being increasingly used in the management of postmenopausal women. However, as clinical trials have demonstrated a significantly increased risk of breast cancer with oral combined estrogen progestin therapy, there is a need to ascertain the risk of including testosterone in such regimens. OBJECTIVE: Evaluation of experimental and epidemiological studies pertaining to the role of testosterone in breast cancer. DESIGN: Literature review. SETTING: The Jean Hailes Foundation, Research Unit. MAIN OUTCOME MEASURES: Mammary epithelial proliferation, apoptosis and breast cancer. RESULTS: In experimental studies, testosterone action is anti proliferative and pro-apoptotic, and mediated via the AR, despite the potential for testosterone to be aromatized to estrogen. Animal studies suggest that testosterone may serve as a natural, endogenous protector of the breast and limit mitogenic and cancer promoting effects of estrogen on mammary epithelium. In premenopausal women, elevated testosterone is not associated with greater breast cancer risk. The risk of breast cancer is also not increased in women with polycystic ovary syndrome who have chronic estrogen exposure and androgen excess. However, in postmenopausal women, who are oestrogen deplete and have increased adipose aromatase activity, higher testosterone has been associated with greater breast cancer risk. CONCLUSION: Available data indicate the inclusion of testosterone in estrogen-progestin regimens has the potential to ameliorate the stimulating effects of hormones on the breast. However, testosterone therapy alone cannot be recommended for estrogen deplete women because of the potential risk of enhanced aromatisation to estrogen in this setting. PMID- 15531123 TI - An evaluation of the contents and quality of menopause information on the World Wide Web. AB - OBJECTIVE: To evaluate the content and quality of currently available World Wide Web based information about menopause. DESIGN: A sample of the 100 top sites obtained with the search engine Google for the key word "menopause" was evaluated according to predefined general and specific criteria, content type, language, and quality. The Internet popularity was established by the number of links to each website. Using a systematic scoring tool each site was assessed on factual information provided and quality of site. RESULTS: The websites studied were heterogeneous in content and quality. The most frequent type of website has commercial content with low quality, biased or useless information. Few sites provided comprehensive medical information about menopause. The scientific societies not always provided complete information about the possible adverse events related with hormone replacement therapy. The results of the Women's Health Initiative (WHI) and the Million Women (MW) studies are discussed by very few websites. Internet popularity did not correlate with measures of quality such as display of authorship, attribution or references, currency of information, and disclosure. CONCLUSION: The content and quality of the websites concerning menopause are widely varied and sometimes biased to commercial goals. The dominant medical information is of low quality and do not comply with general quality scores although the most informative sites have comprehensive content about the menopause including the recent results of the WHI and MW studies. PMID- 15531124 TI - The changing nature of women's sexual health concerns through the midlife years. AB - OBJECTIVE: Information about the sexual health care needs for midlife women is limited. This study compares and contrasts the nature and prevalence of sexual concerns for women as they progress through life and into menopause and the interest and experience these women have in discussing sexual concerns with their physicians. METHODS: Questionnaires were mailed to 2073 eligible women at military medical clinics in the early 1990s. Main outcome measures are self reported sexual concerns and interest and experience in discussing these concerns with their physicians. RESULTS: Over 98% of women reported one or more sexual concerns. Type and intensity of sexual concerns changed as women aged. Most women had not had the topic of sexual health ever raised by their physicians. CONCLUSIONS: The sexual health concerns of women change as they age. Women desire to address their sexual health care needs with their physicians. Physicians should be aware of the common sexual concerns of women, and be comfortable in initiating discussion to address them. PMID- 15531125 TI - Self-reported urogenital symptoms in postmenopausal women: Women's Health Initiative. AB - OBJECTIVE: To examine the prevalence and correlates of self-reported urogenital symptoms (dryness, irritation or itching, discharge, dysuria) among postmenopausal women aged 50-79. DESIGN: A cross-sectional analysis based on n=98,705 women enrolled in the US-based Women's Health Initiative observational study and clinical trials. Urogenital symptoms, symptom severity (mild, moderate, severe), and all covariates were self-reported through questionnaires at enrollment. Prevalence rates of each urogenital symptom were examined and logistic regression was used to identify potential correlates. RESULTS: Prevalence rates for each symptom were: dryness, 27.0%; irritation or itching, 18.6%; discharge, 11.1%; and dysuria, 5.2%. Four factors were correlated with two or more symptoms: Hispanic ethnicity (adjusted odds ratio (AOR)=2.1-3.1 versus white women across all symptoms), obesity (AOR=2.2 severe discharge versus none, AOR=3.6 severe irritation/itching versus none), treated diabetes (pills or shots) compared to no diabetes (AOR=2.4 severe dysuria versus none, AOR=3.2 severe irritation/itching versus none), and vaginal cream HRT/ERT compared to those who never used HRT/ERT (AOR=4.4 severe dryness versus none, AOR=4.6 severe irritation/itching versus none). Factors not associated with the symptoms included sexual activity, age, years since menopause, current smoking, marital status, gravidity, and natural versus surgical menopause. CONCLUSIONS: This is the first report to document urogenital symptoms by race/ethnicity among an exclusively postmenopausal population. We found an elevated prevalence of urogenital symptoms among women who are Hispanic, obese, and/or diabetic. Confirmation of our findings in these subgroups, and, if confirmed, analysis on why these populations are at greater risk, are areas for future research. PMID- 15531126 TI - Progestins used in hormonal replacement therapy display different effects in coronary arteries from New Zealand white rabbits. AB - OBJECTIVES: The aim of this study was in an animal model to assess the vascular effects of different progestins commonly used in hormonal replacement treatment. METHODS: Fifty-six non-atherosclerotic, ovariectomized New Zealand white rabbits were randomized into seven groups: (1) medroxyprogesterone acetate (MPA), (2) norethisterone acetate (NETA), (3) conjugated equine estrogens (CEE), (4) 17-beta estradiol (E2), (5) MPA+CEE , (6) NETA+E2 , (7) or placebo (n=8) and given hormonal treatment through the diet for 4 weeks. Ring segments from the left proximal coronary artery and from the distal part of the left anterior descending coronary artery were microdissected and mounted for isometric tension recordings in a myograph. The vasoconstrictory responses induced by potassium, endothelin-1, calcium and Nw-nitro-L-arginine methyl ester, and the vasodilatory response induced by acetylcholine and sodiumnitroprusside were investigated. The maximum contraction/relaxation (Emax) and the concentration required to induce half the maximum response (EC50) were determined. EC50 values were expressed as the negative logarithm to the molar concentration, pD2=-log EC50. RESULTS: Treatment with MPA alone caused when compared to treatment with NETA an increase in tension development in the distal coronary artery after the addition of potassium ( 6.36+/-0.36 versus 4.31+/-0.42 P<0.005) (single dose response, mN/mm, mean+/ S.E.M.) and endothelin-1 (9.41+/-0.82 versus 6.43+/-0.73 P<0.05) (Emax, mN/mm, mean+/-S.E.M.). Treatment with MPA compared to placebo caused an endothelin-1 induced increase of Emax in the distal coronary artery (9.21+/-0.87 versus 6.51+/ 0.65 P<0.05) and a calcium induced increase of pD2 in both coronary arteries (2.98+/-0.19 versus 2.42+/-0.12 P<0.05, proximal coronary artery) (3.26+/-0.09 versus 2.9+/-0.1 P<0.05, distal coronary artery) (pD2, mean+/-S.E.M.). Treatment with NETA compared to placebo in the proximal coronary artery, after the addition of sodiumnitroprusside caused a decrease of pD2 (5.33+/-0.19 versus 5.94+/-0.13 P<0.05). Treatment with E2 compared to treatment with CEE in the proximal coronary artery caused a decrease of pD2 after the addition of sodiumnitroprusside (5.00+/-0.16 versus 5.77+/-0.28 P<0.05). No significant differences were found between MPA+CEE and NETA+E2. CONCLUSION: Treatment with MPA alone seems to enhance the contractile response to potassium and endothelin-1 in the distal coronary artery compared to NETA, indicating that different progestins used in hormonal replacement treatment may display different effects on contractile functions of coronary arteries. PMID- 15531127 TI - Effects of progestins on estrogen-induced increase in C-reactive protein in postmenopausal women. AB - BACKGROUND: C-reactive protein (CRP) represents an independent risk factor for coronary disease and stroke. Because oral estrogens increase CRP levels, with inflammatory and thrombotic consequences, we determined whether the co administration of a progestin might modify the estrogenic effect on CRP. METHODS: In a non-randomized, non-blinded study, we measured C-reactive protein serum concentrations with high-sensitivity technique (hs-CRP) in 163 healthy postmenopausal women divided into groups as follow: 52 not taking hormones (referent group), and 111 taking hormone replacement therapy (HRT) (42 of whom treated with unopposed estrogen, and 69 with an estrogen/progestin combination). RESULTS: Compared with non-users of hormones, median CRP levels were 66% (95% confidence interval: from 44 to 89%) higher and 112% (95% confidence interval: from 89 to 168%) higher among women using a combined estrogen/progestin regimen and, respectively, among women taking unopposed estrogen [1.54 mg/L in the referent group; 2.56 mg/L in the estrogen/progestin group (P=0.032), and 3.27 mg/L in the unopposed estrogen group (P=0.004)]. Furthermore, there was no difference in CRP distributions between women taking different types of progestins. CONCLUSION: concurrent progestin administration may attenuate estrogen's pro-inflammatory effects, independently on the type of used progestin. PMID- 15531128 TI - Age at natural menopause in Iran. AB - OBJECTIVES: Estimation of mean and median age at menopause among Iranian women using a cumulative distribution by region. METHODS: The number of 8194 women aged between 30 and 65+ who enrolled in National Health Survey in the year 2000 in Iran through cluster sampling were studied and according to their age and the onset of their menopause (12 month consecutive menstrual cessation), the pattern of age at natural menopause has been described using a cumulative distribution. RESULTS: The median age of menopause has been calculated to be 49.9, 49.2, and 49.6 years in urban, rural, and total population, respectively. While the mean age at menopause in total population was 50.4 years (S.D.=4.3). As it has been shown, the median age of menopause in rural areas (49.2) is earlier than urban areas (49.9). This difference is statistically significant (95% CI: 1.00066, 1.03363). CONCLUSION: There are different factors influencing age at natural menopause but there are not certain and homogenous results about those factors in different studies. Therefore, it is obvious that exploring these factors in different regions in Iran merit to be done. PMID- 15531129 TI - Hormonal replacement therapy after stem cell transplantation. AB - OBJECTIVE: To evaluate HRT compliance and efficacy in the treatment of symptomatic ovarian failure in pre-menopausal women after stem cell transplantation (SCT) for malignancies. METHODS: Thirty-one females were selected and prospectively followed in a university bone marrow transplantation unit and gynecologic outpatient clinic in a university teaching hospital. The patients received regular gynecological examinations, hormonal assessment every 6 months including plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), 17-beta estradiol (E2), and transvaginal pelvic ultrasonography, mammography, and computerized bone mineralometry every 12 months. Self-assessment form of menopausal symptoms perception was filled in by all patients before HRT and during the observation period. RESULTS: All patients developed gonadal failure after SCT. The menopausal symptoms more frequently reported were: vasomotor (90%), muscleskeletal symptoms (61%), vulvo-vaginal atrophy (54%), and mood changes (54%). Fifteen premenopausal women out of 31 (51.6%) received systemic HRT on the basis of age (<45 years), absence of medical contraindication or subjective refusal, and complete remission of underlying malignancies post SCT. The remaining patients (48.4%) did not receive HRT mainly for patient's refusal (29%), relapse of malignancy (13%) or liver toxicity (9.6%). HRT was effective in HRT treated patients promptly relieving unpleasant symptoms of gonadal failure. HRT was tolerated without minimal complications or serious side effects. CONCLUSIONS: Dramatic improvement in menopausal symptoms was observed in women on HRT although treatment was feasible only in nearly half of the patients. HRT alleviates unnecessary discomfort and improves the well-being of female patients post-transplant also by preventing menopause related complications. Feasibility and patients' compliance of HRT should be carefully evaluated in long term survivors after stem cell transplantation. PMID- 15531130 TI - The effects of postmenopausal Vitamin D treatment on vaginal atrophy. AB - OBJECTIVES: To evaluate the efficacy of Vitamin D for relieving vaginal atrophy. METHODS: In our cross-sectional study, 60 eligible women were consecutively identified among patients who were followed in physical medicine and rehabilitation's outpatient clinic, during the period March June 2001. The sample consisted of 30 postmenopausal women on 1,25-dihydroxy Vitamin D (calcitriol) treatment, 30 postmenopausal women taking neither hormone replacement nor osteoporosis treatment. Vaginal atrophy was determined by symptoms, physical examination and maturation index (MI). RESULTS: The mean physical findings score in Vitamin D treatment (VDT) group was significantly lower than the mean physical findings score in the group without treatment. Symptom scores were lower in both groups and there was not significantly difference between two groups. As maturation indices: in VDT group, superficial cells proportion was significantly higher and basal, parabasal cells proportion was lower than in the group without treatment. CONCLUSION: The results of current study seem to indicate that vaginal tissue may be a new target organ for therapeutically applied Vitamin D analogues. PMID- 15531131 TI - Continuation rate of hormone replacement therapy in Hong Kong public health sector. AB - OBJECTIVES: To assess the 1-year continuation rate of HRT prescribed in Hong Kong public health sector and to identify factors affecting this continuation rate. METHODS: All women who received at least one dispensed prescription of estrogens between January 1998 and December 2000 from 36 specialist outpatient clinics of the Hospital Authority were selected, and observed for at least 2 years and at most 3 years. The duration of use and variables including age, types of hormones, routes of delivery, dose of estrogen, and prescribing specialty were retrieved from the central prescription database of the Hospital Authority. RESULTS: Of 12,711 incident users of HRT, more than half were aged 50-59. Most (78.5%) of the users took conjugated equine estrogens (CEE) 0.625 mg or related products. Only a small proportion (3.0%) of women used CEE 0.3 mg. Initial estrogen prescriptions were written by gynaecologists in 86.7%. The overall 1-year continuation rate was 68.3%. The highest and lowest continuation rates were observed in women aged 40 49 and the two extreme age groups (35-39 and 70-79), respectively. Better continuation rate was observed in women taking estrogen-only therapy such as CEE or estradiol (overall 76.3%) than in women using continuous combined therapy (58.6%), sequential combined therapy (64.8%), or transdermal estrogen (60.6%). In the age group 60-69, the use of CEE 0.3 mg was associated with better continuation rate than CEE 0.625 mg. CONCLUSIONS: Better continuation rate at 1 year was associated with age younger than 60, oral route of HRT and hysterectomy. PMID- 15531132 TI - Distribution of galanin and galanin transcript in the brain of a galanin overexpressing transgenic mouse. AB - The distribution of galanin mRNA-expressing cells and galanin-immunoreactive (IR) cell bodies and processes was studied in the brain of mice overexpressing galanin under the PDGF-B promoter (GalOE mice) and of wild type (WT) mice, both in colchicine-treated and non-treated animals. In this abstract, we only describe the results in GalOE mouse. A widespread ectopic expression of galanin (both mRNA and peptide) was found, that is a situation when neither transcript nor peptide could be seen in WT mice, not even after colchicine treatment. However, in some regions, such as claustrum, basolateral amygdala, thalamus, CA1 pyramidal cells, and Purkinje cells only galanin mRNA could be detected. In the forebrain galanin was seen in the mitral cells of the olfactory bulb, throughout the cortex, in the basolateral amygdaloid nucleus, claustrum, granular and pyramidal cell layers of the hippocampus, subiculum and presubiculum. In the thalamus, the anterodorsal, mediodorsal, intermediodorsal and mediodorsal lateral nuclei, the reuniens and reticular nuclei showed ectopic expression of galanin. Within the hypothalamus, neurons of the suprachiasmatic nucleus contained galanin. In the mesencephalon, the geniculate nucleus, nucleus ruber, the mesencephalic trigeminal and reticulotegmental nuclei ectopically expressed galanin. In the cerebellum, galanin was observed in the Purkinje cells and in the lateral and interposed cerebellar nuclei. In the pons, sensory and motor nuclei of the trigeminal nerve, the laterodorsal and dorsal tegmental nuclei, the pontine, reticulotegmental and gigantocellular reticular nuclei expressed galanin. Within the medulla oblongata, labeled cells were detected in the facial, ambiguus, prepositus, lateral paragigantocellular and lateral reticular nuclei, and spinal trigeminal nucleus. High densities of galanin-IR fibers were found in the axonal terminals of the lateral olfactory tract, the hippocampal and presumably the cerebellar mossy fibers system, in several thalamic and hypothalamic regions and the lower brain stem. Possible functional consequences of galanin overexpression are discussed. PMID- 15531133 TI - Effects of intracerebroventricular infusion of genistein on gonadotrophin subunit mRNA and immunoreactivity of gonadotrophins and oestrogen receptor-alpha in the pituitary cells of the anoestrous ewe. AB - The present study was designed to demonstrate whether genistein, a synthetic phytoestrogen, infused into the third ventricle of the brain could affect the gonadotrophic cells regarding the presence of oestrogen receptor-alpha immunoreactivity and gonadotrophin subunit mRNA hybridising reaction in the ewe. Ewes (n=7), aged 2 years, in early anoestrous season were infused with Ringer Locke solution (control, n=3) or 10 microg/100 microl/h of genistein (n=4) into the third ventricle over a 5 h period and slaughtered the following morning. Immunoreactivity of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and oestrogen receptor-alpha (ERalpha) was determined in the adenohypophysis by immunohistochemistry using antibodies raised against LHbeta, FSHbeta, and ERalpha. Messenger RNA analyses were performed by non-isotope in situ hybridisation using sense and antisense riboprobes produced from beta subunits of LH and FSH cDNA clones. Computer image analysis was used to determine the percent of cells exhibiting immunohistochemical and/or hybridising reaction. It was found that in ewes infused with genistein, the percentage of LH-positive cells and the density of immunoreactive-LHbeta material decreased significantly (P10 mm in the short axis) and pleural changes (including effusion, irregular thickening, or nodularity) as the cardinal imaging findings of lung metastases. A five-point scoring system was designed to indicate the probability of lung metastasis from primary gynecologic malignancies. The five-point scores of DR, LDCT, and SDCT were analyzed by receiver operating characteristic (ROC) curve. RESULTS: SDCT probability scores of +2 and -2 were set to indicate true positive and true negative for pulmonary nodule, mediastinal lymphadenopathy, and pleural effusion, respectively. All the areas under the ROC curve of LDCT appeared to be larger than those of DR[pulmonary nodule: 0.96 [95% confidence interval (CI): 0.92-1.01] vs. 0.74 [95% CI: 0.57-0.91], 0.82 [95% CI: 0.70-0.95] vs. 0.61 [95% CI: 0.50 0.77]; mediastinal lymphadenopathy: 0.98 [95% CI: 0.93-1.03] vs. 0.90 [95% CI: 0.79-1.01], 0.94 [95% CI: 0.82-1.06] vs. 0.66 [95% CI: 0.44-0.88]; and pleural effusion: 0.98 [95% CI: 0.93-1.03] vs. 0.56 [95% CI: 0.29-0.82], 0.90 [95% CI: 0.74-1.05] vs. 0.46 [95% CI: 0.23-0.68]]. CONCLUSION: The performance of LDCT were comparable to those of SDCT and superior to those of DR for detection of pulmonary nodule, mediastinal lymphadenopathy, and pleural effusion. By using LDCT, there was no need of intravenous contrast injection and less radiation exposure. We propose a protocol including standard-dose abdominal CT and low-dose chest CT for the initial and follow-up stagings of primary gynecologic malignancy. The use of chest DR is unnecessary. PMID- 15531141 TI - Neurofibroma of the breast in a boy with neurofibromatosis type 1. AB - Neurofibromas occurring in the breast are very rare. A 10-year-old boy with neurofibromatosis type 1 (NF1) presented with a palpable mass on his left breast. US showed a well-defined and hypoechoic solid mass within the subcutaneous fat tissue in the areolar area. Mass was isointense compared to muscle and hypointense compared to fat tissue on TIW and T2W MR images. Excisional biopsy was applied to the mass. The diagnosis of gynecomastia and neurofibroma was made on the basis of histopathological and immunohistochemical findings. PMID- 15531143 TI - Evaluation of perforated and nonperforated appendicitis with CT. AB - Fifty-three patients with 38 cases of perforated appendicitis and 15 cases of appendicitis without perforation were evaluated based on the computed tomography (CT) appearances of appendiceal diameter, phlegmon, abscess, extraluminal air, appendiceal wall enhancement, lateroconal fascial thickening, appendicolith, bowel wall thickening, ascites, ileal wall enhancement, peritoneal enhancement, periappendiceal fluid, omental haziness, retrocecal appendix, intraluminal air, and the combination of intraluminal air and appendicolith. The result of appendiceal diameter was compared using two-sample Student's t test, and the other CT findings were analyzed by Fisher's Exact Test. Our results showed that appendix was larger in caliber in perforated appendix (P< .05). Direct CT signs (i.e., phlegmon, abscess, and extraluminal air) were more specific for perforated appendicitis (P< .05). Indirect signs (bowel wall thickening, ascites, ileal wall enhancement, intraluminal air, and combined intraluminal air and appendicolith) were also found in higher incidence in appendiceal perforation (P< .05). Appendiceal enlargement and ileal wall enhancement were the two predominant findings in one case of perforation. We concluded that direct and indirect CT appearances can differentiate appendicitis with and without perforation. Indirect signs may be helpful in difficult case. PMID- 15531142 TI - Radiology-Pathology Conference: Calcified untreated gastric cancer. AB - Adenocarcinoma of the stomach represents upwards of 95% of all gastric malignancy. Calcifications in untreated gastric cancer are rare, but typically represent mucin-producing tumor. We report the radiology and pathology of a patient who presented with gastric wall calcifications on CT and a mucin producing adenocarcinoma at pathologic examination of a gastric biopsy specimen. PMID- 15531144 TI - Barium enema "shoulder" sign: a useful predictor of pericolonic fat invasion in colorectal cancer. AB - PURPOSE: We aimed to determine the presence of the "shoulder" sign in prediction of pericolonic fat invasion on double-contrast barium enema (DCBE) studies in colorectal cancer. MATERIALS AND METHODS: Patients at our institution with apparent colorectal cancer were identified from a computer search of both radiological and surgical pathology records during a recent 30-month period. The presence of a shoulder sign was defined as a tumor margin having an abrupt transition from the intraluminal component to the adjacent normal mucosa, forming a shouldered edge on barium studies. RESULTS: A total of 49 patients (53 lesions) for whom the DCBE studies and surgical pathology records were available for review were proven to have colorectal carcinomas. DCBE studies revealed that 45 patients with lesions (85%) presented with a "shoulder" sign. All of these lesions were correctly diagnosed as colorectal cancer. Of these, 89% had pericolonic fat invasion and 44% had regional lymph node metastases. Of lesions without a shoulder sign, 25% had pericolonic fat invasion and 25% had nodal metastases. CONCLUSION: The presence of shoulder sign detected with barium enema can be a useful predictor of pericolonic fat invasion in colorectal cancer. PMID- 15531145 TI - Preoperative local staging of rectal cancer with endorectal MR imaging: comparison with histopathologic findings. AB - The purpose of this prospective study was to assess the accuracy of endorectal MR imaging in the preoperative local staging of rectal cancers. In 20 cases, we correlated endorectal MR imaging findings with postoperative histopathologic staging according to TNM classification. The accuracy of endorectal MR for determining the T stage of rectal cancer was 85%. The sensitivity and specificity for detecting lymph node metastases were 90.9% and 55.5%, respectively. PMID- 15531146 TI - Deep venous thrombosis: comparison of indirect multidetector CT venography and sonography of lower extremities in 26 patients. AB - OBJECTIVE: To compare the accuracy of indirect mutidetector row computed tomographic (MDCT) venography with lower extremity venous sonography for the diagnosis of femoropopliteal deep venous thrombosis (DVT), and to determine the frequency and location of DVT at MDCT venography. MATERIALS AND METHODS: Twenty six consecutive patients suspected of having pulmonary embolism (PE) underwent both combined MDCT venography and MDCT pulmonary angiography and lower extremity venous sonography. Indirect MDCT venography was acquired from the upper calves to the mid-abdomen following MDCT pulmonary angiography. The CT venographic findings were compared with those of sonography for the diagnosis of femoropopliteal DVT. All CT scans were also reviewed for the frequency and location of DVT. RESULTS: Indirect MDCT venography disclosed DVT in 19 patients, and 12 of whom also had PE. Seventeen patients with thrombosis in the femoropopliteal veins were identified in both indirect MDCT venography and sonography. The sensitivity and specificity of indirect MDCT venography for femoropopliteal DVT, as compared with sonography, were both 100%. In one patient DVT in the superficial femoral vein was detected using only indirect MDCT venography. MDCT venography also showed superior extension of femoropopliteal DVT to the inferior vena cava and iliac veins in four patients and thrombosis isolated to the inferior vena cava and common iliac vein thrombosis in one patient. CONCLUSIONS: Indirect MDCT venography is as accurate as sonography in the diagnosis of femoropopliteal DVT. MDCT venography can further reveal thrombus in large pelvis veins and the inferior vena cava, an important advantage over sonographic screening for DVT. PMID- 15531147 TI - Adaptive systems and hybrid computational intelligence in medicine. PMID- 15531148 TI - Adaptive ventilator FiO2 advisor: use of non-invasive estimations of shunt. AB - A non-invasive and simple method of parameter estimation has been developed for the model-based decision support of the artificial ventilation in intensive care units. The parameter concerned was the respiratory shunt. Originally, the shunt had to be estimated using a numerical algorithm, which was slow and unreliable. The estimation process also required the knowledge of other parameters, whose values could only be obtained using invasive monitoring equipment. In this paper, the respiratory index is used for the shunt estimation. A linear regression model and a non-linear adaptive neuro-fuzzy inference system (ANFIS) model were used to describe the relationship between the respiratory index and the shunt. The shunts estimated using these models were then used to calculate the fractional inspired oxygen needed to attain the target arterial oxygen level of the model patient. The advisor also utilises population median values of the cardiac index and oxygen consumption index. This alleviates the need for invasive monitoring. In a simulation study, the mean squared error of the control using the ANFIS model was 0.75 kPa2 compared to 2.06 kPa2 using the linear regression model. Therefore, the performance of the FiO2 advisor was better when the shunt was estimated using the non-linear ANFIS model. PMID- 15531149 TI - Evaluation of glucose controllers in virtual environment: methodology and sample application. AB - OBJECTIVE: Adaptive systems to deliver medical treatment in humans are safety critical systems and require particular care in both the testing and the evaluation phase, which are time-consuming, costly, and confounded by ethical issues. The objective of the present work is to develop a methodology to test glucose controllers of an artificial pancreas in a simulated (virtual) environment. MATERIAL AND METHODS: A virtual environment comprising a model of the carbohydrate metabolism and models of the insulin pump and the glucose sensor is employed to simulate individual glucose excursions in subjects with type 1 diabetes. The performance of the control algorithm within the virtual environment is evaluated by considering treatment and operational scenarios. RESULTS: The developed methodology includes two dimensions: testing in relation to specific life style conditions, i.e. fasting, post-prandial, and life style (metabolic) disturbances; and testing in relation to various operating conditions, i.e. expected operating conditions, adverse operating conditions, and system failure. We define safety and efficacy criteria and describe the measures to be taken prior to clinical testing. The use of the methodology is exemplified by tuning and evaluating a model predictive glucose controller being developed for a wearable artificial pancreas focused on fasting conditions. CONCLUSION: Our methodology to test glucose controllers in a virtual environment is instrumental in anticipating the results of real clinical tests for different physiological conditions and for different operating conditions. The thorough testing in the virtual environment reduces costs and speeds up the development process. PMID- 15531150 TI - Prediction of visual perceptions with artificial neural networks in a visual prosthesis for the blind. AB - Within the framework of the OPTIVIP project, an optic nerve based visual prosthesis is developed in order to restore partial vision to the blind. One of the main challenges is to understand, decode and model the physiological process linking the stimulating parameters to the visual sensations produced in the visual field of a blind volunteer. We propose to use adaptive neural techniques. Two prediction models are investigated. The first one is a grey-box model exploiting the neurophysiological knowledge available up to now. It combines a neurophysiological model with artificial neural networks, such as multi-layer perceptrons and radial basis function networks, in order to predict the features of the visual perceptions. The second model is entirely of the black-box type. We show that both models provide satisfactory prediction tools and achieve similar prediction accuracies. Moreover, we demonstrate that significant improvement (25%) was gained with respect to linear statistical methods, suggesting that the biological process is strongly non-linear. PMID- 15531151 TI - Evolving rule-based systems in two medical domains using genetic programming. AB - OBJECTIVE: To demonstrate and compare the application of different genetic programming (GP) based intelligent methodologies for the construction of rule based systems in two medical domains: the diagnosis of aphasia's subtypes and the classification of pap-smear examinations. MATERIAL: Past data representing (a) successful diagnosis of aphasia's subtypes from collaborating medical experts through a free interview per patient, and (b) correctly classified smears (images of cells) by cyto-technologists, previously stained using the Papanicolaou method. METHODS: Initially a hybrid approach is proposed, which combines standard genetic programming and heuristic hierarchical crisp rule-base construction. Then, genetic programming for the production of crisp rule based systems is attempted. Finally, another hybrid intelligent model is composed by a grammar driven genetic programming system for the generation of fuzzy rule-based systems. RESULTS: Results denote the effectiveness of the proposed systems, while they are also compared for their efficiency, accuracy and comprehensibility, to those of an inductive machine learning approach as well as to those of a standard genetic programming symbolic expression approach. CONCLUSION: The proposed GP-based intelligent methodologies are able to produce accurate and comprehensible results for medical experts performing competitive to other intelligent approaches. The aim of the authors was the production of accurate but also sensible decision rules that could potentially help medical doctors to extract conclusions, even at the expense of a higher classification score achievement. PMID- 15531153 TI - Evolutionary impact of human Alu repetitive elements. AB - Early studies of human Alu retrotransposons focused on their origin, evolution and biological properties, but current focus is shifting toward the effect of Alu elements on evolution of the human genome. Recent analyses indicate that numerous factors have affected the chromosomal distribution of Alu elements over time, including male-driven insertions, deletions and rapid CpG mutations after their retrotransposition. Unequal crossing over between Alu elements can lead to local mutations or to large segmental duplications responsible for genetic diseases and long-term evolutionary changes. Alu elements can also affect human (primate) evolution by introducing alternative splice sites in existing genes. Studying the Alu family in a human genomic context is likely to have general significance for our understanding of the evolutionary impact of other repetitive elements in diverse eukaryotic genomes. PMID- 15531154 TI - Homing endonuclease genes: the rise and fall and rise again of a selfish element. AB - Homing endonuclease genes (HEGs) are selfish genetic elements that spread by first cleaving chromosomes that do not contain them and then getting copied across to the broken chromosome as a byproduct of the repair process. The success of this strategy will depend on the opportunities for homing--in other words, the frequency with which HEG(+) and HEG(-) chromosomes come into contact--which varies widely among host taxa. HEGs are also unusual in that the selection pressure for endonuclease function disappears if they become fixed in a population, which makes them susceptible to degeneration and imposes a need for regular horizontal transmission between species. HEGs will be selected to reduce the harm done to the host organism, and this is expected to influence the evolution of their sequence specificity and maturase functions. HEGs may also be domesticated by their hosts, and are currently being put to human uses. PMID- 15531155 TI - Start-up entities in the origin of new genes. AB - The remarkable diversity in the contents of genomes raises questions about how new genes and new functions originate. Recent evidence indicates that parasitism, particularly the molecular interactions between phage and their bacterial hosts, is a likely mechanism for generating new genes. This invention of such novel functions seems to be founded on a strategy that secures the short-term survival of parasitic elements and thereby contributes to the renovation of gene repertoires in their host. PMID- 15531156 TI - Evolution of genome size: multilevel selection, mutation bias or dynamical chaos? AB - In the past two years, new data on conceptual aspects of the evolution of eukaryotic genome size have appeared, including the adaptivity of genome enlargement, the mechanisms of genome size change and the relation of genome size to organismal complexity. New data on the hypotheses of "selfish DNA" and "mutational equilibrium" have been recently obtained. A relationship is emerging between the intragenomic distribution of noncoding DNA and differential gene expression, which suggests that noncoding DNA is involved in epigenetic organization of the genome and organismal complexity. The standpoint of dynamical chaos, which integrates multilevel selection and mutation biases, may provide a framework for studying the evolution of genome size. PMID- 15531157 TI - Genomic changes following host restriction in bacteria. AB - Many genomic sequences have been recently published for bacteria that can replicate only within eukaryotic hosts. Comparisons of genomic features with those of closely related bacteria retaining free-living stages indicate that rapid evolutionary change often occurs immediately after host restriction. Typical changes include a large increase in the frequency of mobile elements in the genome, chromosomal rearrangements mediated by recombination among these elements, pseudogene formation, and deletions of varying size. In anciently host restricted lineages, the frequency of insertion sequence elements decreases as genomes become extremely small and strictly clonal. These changes represent a general syndrome of genome evolution, which is observed repeatedly in host restricted lineages from numerous phylogenetic groups. Considerable variation also exists, however, in part reflecting unstudied aspects of the population structure and ecology of host-restricted bacterial lineages. PMID- 15531158 TI - Sex chromosome evolution in non-mammalian vertebrates. AB - Birds, reptiles, amphibia and fish have an enormous variety of chromosomal sex determination mechanisms that apparently do not follow any phylogenetic or taxonomic scheme. A similar picture is now emerging at the molecular level. Most genes that function downstream of the mammalian master sex-determining gene, Sry, have been found in non-mammalian vertebrates. Although the components of the machinery that determines sex seem to be conserved, their interaction and most importantly the initial trigger is not the same in all vertebrates. This variety is the consequence of the extremely dynamic process of the evolution of sex determination mechanisms and sex chromosomes, which is prone to create differences rather than uniformity. PMID- 15531159 TI - A platypus' eye view of the mammalian genome. AB - The genome of monotremes, like the animals themselves, is unique and strange. The importance of monotremes to genomics depends on their position as the earliest offshoot of the mammalian lineage. Although there has been controversy in the literature over the phylogenetic position of monotremes, this traditional interpretation is now confirmed by recent sequence comparisons. Characterizing the monotreme genome will therefore be important for studying the evolution and organization of the mammalian genome, and the proposal to sequence the platypus genome has been received enthusiastically by the genomics community. Recent investigations of X-chromosome inactivation, genomic imprinting and sex chromosome evolution provide good examples of the power of the monotreme genome to inform us about mammalian genome organization and evolution. PMID- 15531160 TI - Comparative primate genomics: the year of the chimpanzee. AB - This is the year of the chimpanzee genome. Chimpanzee chromosome 22 has been sequenced and soon will be followed by the whole genome, and thousands of chimpanzee cDNA sequences are available for comparative analysis. Not only does this genomic information allow us to identify human-specific changes in particular genes that are potentially under selection, but also to understand molecular evolutionary dynamics characterizing the two most closely related mammalian genomes sequenced so far. Studies comparing gene expression in chimpanzees and other closely related primates reveal significant species differences in brain, liver and fibroblasts. New empirical data, in combination with models of speciation, are giving insight into how humans and chimpanzees speciated. PMID- 15531161 TI - The evolution of eutherian chromosomes. AB - The gross organization of the genome of Eutheria (placental mammals) into chromosomes follows a simple architecture that, with some minor changes, is almost completely conserved for more than 100 million years in various species of almost all extant mammalian orders. Recent molecular cytogenetic results- especially those from the assumed oldest clade, the Afrotheria--suggest an ancestral karyotype that would calculate the "default" frequency of gross rearrangements to less than two changes within 10 million years of mammalian evolution. The main changes are the fission, movement and subsequent fusion of large chromosome segments or of chromosome arms. Reciprocal translocations are the exception. Chromosome numbers may have increased or decreased significantly in this fusion/fission process but, in most instances, the main architecture still remains evident. There are, however, some exceptions in mammals with extremely derived karyotypes. PMID- 15531162 TI - A structured ancestral population for the evolution of modern humans. AB - The view that modern humans evolved through a bottleneck from a single founding group of archaic Homo is being challenged by new analyses of contemporary genetic variation. A wide range of middle to late Pleistocene ages for gene genealogies and evidence for early population structures point to a diverse and scattered ancestry associated with a metapopulation history of local extinctions, re colonization and admixture. A different balance of the same processes has shaped chimpanzee diversity. PMID- 15531163 TI - Speciation genes. AB - Until recently, the genes that cause reproductive isolation remained black boxes. Consequently, evolutionary biologists were unable to answer several questions about the identities and characteristics of "speciation genes". Over the past few years, however, evolutionary geneticists have finally succeeded in isolating several such genes, providing our first glimpse at factors that are thought to be representative of those underlying the origin of species. Evolutionary analysis of these genes suggests that speciation results from positive Darwinian selection within species. Molecular evolutionary study of the genes causing reproductive isolation may represent an important new phase in the study of speciation. PMID- 15531164 TI - Eukaryotic gene regulation by targeted chromatin re-modeling at dispersed, middle repetitive sequence elements. AB - RNA interference might have evolved to minimize the deleterious impact of transposable elements and viruses on eukaryotic genomes, because mutations in genes within the RNAi pathway cause mobilization of transposons in nematodes and flies. Although the first examples of RNAi involved post-transcriptional gene silencing, recently the pathway has been shown to act at the transcriptional level. It does so by establishing a chromatin configuration on the target DNA that has many of the hallmarks of heterochromatin, thus preventing its transcription. Members of dispersed, repeated sequence families appear to have been utilized by the RNAi machinery to regulate nearby genes in yeast. The unusual genomic distribution of three repeated element families in the chicken, fruit-fly and nematode genomes prompts speculation that some of these repeats have been co-opted to control gene expression, either locally or over extended chromosomal domains. PMID- 15531165 TI - The value-added genome: building and maintaining genomic cytosine methylation landscapes. AB - Epigenetic marks, such as cytosine methylation and post-translational histone modifications, are important for interpreting and managing eukaryotic genomes. Recent genetic studies in plants have uncovered details on the different interwoven mechanisms that are responsible for specification of genomic cytosine methylation patterns. These mechanisms include targeting cytosine methylation using heterochromatic histone modifications and RNA guides. Genomic cytosine methylation patterns also reflect locus-specific demethylation initiated by specialized DNA glycosylases. While genetics continues to more fully define these mechanisms, genomic studies in Arabidopsis have yielded an unprecedented high resolution view of how epigenetic marks are layered over a genome. PMID- 15531166 TI - Epigenetic germline inheritance. AB - Our increased knowledge of epigenetic reprogramming supports the idea that epigenetic marks are not always completely cleared between generations. Incomplete erasure at genes associated with a measurable phenotype can result in unusual patterns of inheritance from one generation to the next. It is also becoming clear that the establishment of epigenetic marks during development can be influenced by environmental factors. In combination, these two processes could provide a mechanism for a rapid form of adaptive evolution. PMID- 15531167 TI - Progress and challenges in profiling the dynamics of chromatin and transcription factor binding with DNA microarrays. AB - ChIP-chip, or chromatin immunoprecipitation followed by DNA microarray analysis, has proven to be an efficient means of mapping protein-genome interactions. Recent experiments using this tool are beginning to reveal the complex dynamics of transcription factor binding and chromatin organization, and how these processes interact with each other to generate a cellular response to environmental and developmental cues. Data derived from this approach, particularly data involving chromatin components and histone modifications, might be affected by assumptions underlying the procedure, and the data might be made more useful by adoption of standardized whole-genome microarray platforms. PMID- 15531169 TI - Effects of experimentally induced plantar insensitivity on forces and pressures under the foot during normal walking. AB - Pressures under the foot during level walking were measured in 15 healthy young adults (8 females, 7 males, mean age 25.7, S.D. 5.3) before and after immersing the feet in ice-cold water (2 degrees C) for 30 min to evaluate the role of plantar insensitivity on gait patterns. Following ice water immersion, there was a significant decrease in walking speed. Maximum forces and peak pressures under the foot decreased, with the exception of an increase in loading under the third to fifth metatarsal heads. Contact times increased under all regions of the foot, and force-time and pressure-time integrals increased under the second and third to fifth metatarsal head regions. It is concluded that plantar insensitivity significantly alters the distribution, duration, and to a lesser extent, the magnitude of forces and pressures under the foot when walking. These results suggest that in the neuropathic foot, gait changes caused by plantar insensitivity may be partly responsible for the redistribution and altered duration of loading, whereas the increase in the magnitude of forces and pressures are primarily due to other disease-related factors. PMID- 15531170 TI - Secondary gait compensations in individuals without neuromuscular involvement following a unilateral imposed equinus constraint. AB - Ankle equinus is the most commonly identified impairment of individuals with spastic hemiplegia (SH). However, it is not clear how equinus at the ankle may contribute to gait deviations at other joints. The purpose of this study was to determine what compensatory gait deviations may occur as a result of an imposed, unilateral equinus constraint. Gait data were collected on 12 adult subjects with and without one ankle constrained in equinus using a unique taping method. Knee extension at initial contact, knee extension in mid stance, and hip extension at terminal stance were all found to be significantly reduced on the ipsilateral side as a result of the ankle constraint. On the unconstrained or contralateral side, subjects tended to adopt a foot-flat or toe-first initial contact pattern. This study suggests that stance phase limitations in both hip and knee extension in the gait of persons with hemiplegia are not necessarily caused by limited length of the involved side hamstrings and/or hip flexors, but rather that they can occur as the result of an ankle plantarflexor contracture alone. Deviations in the contralateral foot contact pattern can also occur secondary to unilateral equinus and should not be assumed to represent bilateral involvement. PMID- 15531171 TI - Dynamic instability during obstacle crossing following traumatic brain injury. AB - Patients with traumatic brain injury (TBI) complain of "imbalance" or "unsteadiness" while walking, despite a normal gait on clinical examination. Thus, the purpose of this study was to determine if it was possible to quantitatively assess dynamic stability that did not have an obvious neuromuscular origin in individuals following TBI. Ten patients with documented TBI and 10 age, gender, and stature-matched healthy individuals participated in the study. All subjects were instructed to perform unobstructed level walking and to step over obstacles corresponding to 2.5%, 5%, 10%, and 15% of their height. A 13-link biomechanical model of the human body was used to compute the kinematics of the whole body center of mass (COM). Subjects with TBI walked with a significantly slower gait speed and shorter stride length than their matched controls. Furthermore, subjects with TBI displayed a significantly greater and faster medio-lateral (M-L) COM motion and maintained a significantly greater M-L separation distance between their COM and center of pressure (COP) than their matched control subjects. These measurements indicate that subjects with TBI have difficulty maintaining dynamic stability in the frontal plane and have a reduced ability to successfully arrest their sagittal momentum. These findings provide an objective measurement that reflects the complaints of instability not observable on clinical examination for individuals who have suffered a TBI. This ability to identify any functional impairment after a traumatic brain injury that may affect patient safety is critical for prevention of re-injury during the recovery period. PMID- 15531172 TI - Locomotor adaptations for changes in the slope of the walking surface. AB - The goal of this study was to examine the transition of walking from a level surface onto different inclined surfaces. Kinematic data of limb and trunk segments were recorded from individuals as they approached and stepped onto four different ramped surfaces (slopes= 3 degrees , 6 degrees , 9 degrees , 12 degrees ). This transition introduced significant adaptations to the swing limb trajectory that were evident in even the lowest ramp condition and appear to be scaled to the ramp inclination although the nature of this scaling seemed to change between the 6 degrees and 9 degrees conditions. An increased forward pitch of the trunk orientation during all ramp conditions was initiated early on during the preceding stance phase on level ground. The swing limb modification essentially consisted of a two-stage response. The initial response of the limb trajectory changes was not specific to the degree of inclination but later changes were dependent on the ramp condition. The initial response is to ensure a safe toe clearance as the foot approaches the edge of the ramp and then later modifications provide the appropriate positioning of the limb to prepare for an elevated foot contact. Early changes were actively produced through an increased pull-off by the hip flexors and an elevation of the swing limb by the active muscle control of the stance limb. Ankle dorsiflexion also appears to have a supporting role increasing toe clearance. Absorption at the hip and knee during later swing contribute to control and position the limb in preparation for foot contact. These strategies were similar to those adopted for step changes in the level of the walking surface where there are similar demands of the quickly moving the limb forward and upward, however, the positioning of the limb for new angled landing surface requires further adaptations. PMID- 15531173 TI - Assessment and validation of a simple automated method for the detection of gait events and intervals. AB - A simple and rapid automatic method for detection of gait events at the foot could speed up and possibly increase the repeatability of gait analysis and evaluations of treatments for pathological gaits. The aim of this study was to compare and validate a kinematic-based algorithm used in the detection of four gait events, heel contact, heel rise, toe contact and toe off. Force platform data is often used to obtain start and end of contact phases, but not usually heel rise and toe contact events. For this purpose synchronised kinematic, kinetic and video data were captured from 12 healthy adult subjects walking both barefoot and shod at slow and normal self-selected speeds. The data were used to determine the gait events using three methods: force, visual inspection and algorithm methods. Ninety percent of all timings given by the algorithm were within one frame (16.7 ms) when compared to visual inspection. There were no statistically significant differences between the visual and algorithm timings. For both heel and toe contact the differences between the three methods were within 1.5 frames, whereas for heel rise and toe off the differences between the force on one side and the visual and algorithm on the other were higher and more varied (up to 175 ms). In addition, the algorithm method provided the duration of three intervals, heel contact to toe contact, toe contact to heel rise and heel rise to toe off, which are not readily available from force platform data. The ability to automatically and reliably detect the timings of these four gait events and three intervals using kinematic data alone is an asset to clinical gait analysis. PMID- 15531174 TI - Classification of equinus in ambulatory children with cerebral palsy discrimination between dynamic tightness and fixed contracture. AB - In this study a generalised dynamic neural network (GDNN) was designed to process gait analysis parameters to evaluate equinus deformity in ambulatory children with cerebral palsy. The aim was to differentiate dynamic calf muscle tightness from fixed muscle contracture. Patients underwent clinical examination and had instrumented gait analysis before evaluating their equinus under anaesthesia and muscle relaxation at the time of surgery to improve gait. The performance of the clinical examination, the subjective interpretation of gait analysis results, and the application of the neural network to assess ankle function were compared to the examination under anaesthesia. Evaluation of equinus by a Neural Network showed high sensitivity and specificity values with a likelihood ratio of +14.63. The results indicate that dynamic calf muscle tightness can be differentiated from fixed calf muscle contracture with considerable precision that might facilitate clinical decision-making. PMID- 15531175 TI - Postural and respiratory activation of the trunk muscles changes with mode and speed of locomotion. AB - Despite the importance of the deep intrinsic spinal muscles for trunk control, few studies have investigated their activity during human locomotion or how this may change with speed and mode of locomotion. Furthermore, it has not been determined whether the postural and respiratory functions, of which these muscles take part, can be coordinated when locomotor demands are increased. EMG recordings of abdominal and paraspinal muscles were made in seven healthy subjects using fine-wire and surface electrodes. Measurements were also made of respiration and gait parameters. Recordings were made for 10s as subjects walked on a treadmill at 1 and 2 ms(-1) and ran at 2, 3, 4 and 5 ms(-1). Unlike the superficial muscles, transversus abdominis was active tonically throughout the gait cycle with all tasks, except running at speeds of 3 ms(-1) and greater. All other muscles were recruited in a phasic manner. The relative duration of these bursts of activity was influenced by speed and/or mode of locomotion. Activity of all abdominal muscles, except rectus abdominis (RA), was modulated both for respiration and locomotor-related functions but this activity was affected by the speed and mode of locomotion. This study provides evidence that the deep abdominal muscles are controlled independently of the other trunk muscles. Furthermore, the pattern of recruitment of the trunk muscles and their respiratory and postural coordination is dependent on the speed and mode of locomotion. PMID- 15531176 TI - Upper body accelerations during walking in healthy young and elderly men. AB - The purpose of the present study was to assess whether any differences existed in the upper body accelerations of young and elderly subjects during natural speed walking. Head and trunk accelerations in eight young subjects (aged 23+/-4 years) and eight healthy elderly subjects (aged 74+/-3 years) were measured during level walking on a 20 m walkway using a pair of tri-axial accelerometers. Heel contact and toe-off events were determined using a footswitch system embedded in the innersole of the right shoe. Gait measures assessed included; stride, stance and swing durations, cadence, gait velocity, step length and 3D head and trunk accelerations. All acceleration variables were normalised to walking speed before statistical analysis. The main findings of this study were: (1) the peak positive anterior-posterior (AP) trunk acceleration associated with push-off was significantly lower for elderly subjects, (2) the peak negative AP head and trunk accelerations following heel contact was significantly higher for elderly subjects, and (3) the time delay between trunk and head accelerations experienced in the AP direction was significantly lower for the elderly compared to the young group. Together, these results suggest that elderly subjects exhibit different patterns of upper body motion in the direction of travel compared to younger subjects. These differences are probably motivated by the need to maximise dynamic stability during critical parts of the gait cycle. PMID- 15531177 TI - A force measuring treadmill in clinical gait analysis. AB - This preliminary study presents the development and testing of an instrumented treadmill device measuring the ground reaction forces (GRFs) and the feasibility of using this force measuring treadmill (FMT) in clinical gait analysis. A commercially available treadmill was modified and fitted out with three dimensional strain-gauge force transducers. Tests of linearity, centre of pressure position (CoP), cross talk, natural frequency, background noises, and belt speed were undertaken in order to assess the performance of the FMT. In addition, the GRFs and segmental kinematics were recorded while healthy subjects and patients walked on the FMT, in order to compute the net ankle joint moments and the body centre of mass (CMb) kinematics and mechanics. The preliminary results of technical tests were satisfactory with an error less than 10% and dynamic tests in healthy subjects corresponded to the literature. The results of patients were clearly disturbed, demonstrating the ability of the FMT to discriminate pathological gaits from normal ones. We concluded that the GRFs measurements obtained from the FMT seem valid and the clinical assessment of net joint moments and CMb kinematics and mechanics seem feasible. The FMT could be useful device for clinical research and routine gait analysis since it allows gaining some extra room and quickly collecting the GRFs during a large number of successive gait cycles and over a wide range of steady-state gait speeds. However, more work is needed in this area in order to confirm the present results, collect reference data and validate the methodology across pathologies. PMID- 15531178 TI - Stride variability: beyond length and frequency. PMID- 15531180 TI - Non-insulin-dependent diabetes, bone mineral density, and cardiovascular risk factors. AB - People with Type 2 diabetes have bone mass alterations and may have a higher risk of hip fractures. Moreover, they have increased cardiovascular risk factors. The objective of this paper is to investigate the association among non-insulin dependent diabetes, bone mineral density (BMD), and cardiovascular risk factors. Ninety-two patients (36 males and 56 females) were studied and cardiovascular risk factors were measured: total cholesterol, triglycerides, lipoprotein(a), low density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), glucose, HbA1c, and microalbuminuria. The densitometric studies were carried out in the calcaneal region using a DEXA densitometer. The diabetic women had a higher BMD than the control group (0.502 +/- 0.127 vs. 0.408 +/- 0.102, P = .027). The women showed a positive relationship between BMD and triglycerides (r =. 478, P = .0001) and a negative relationship with HDL-C (r = -.322, P = .016). The men had a BMD similar to that of the control group, and there was no relationship with the cardiovascular risk factors. When a multivariate logistic regression analysis was performed with the presence of osteoporosis as a dependent variable and each lipid level, age, sex, and BMI as independent variables, only age and BMI were found to be associated with the presence of osteopososis. The diabetic women had a higher BMD than the controls, and there was no relationship between osteoporosis and cardiovascular risk factors in diabetics. PMID- 15531181 TI - Association between hematological parameters and metabolic syndrome components in a Chinese population. AB - Insulin resistance, an essential core contributing to the metabolic syndrome (MS), has been demonstrated in some studies to be associated with white blood cell (WBC) or red blood cell (RBC) counts. The present study was undertaken to assess systemically the relationship between WBC or RBC counts and various clinical features of MS in a large Chinese population at Taiwan. A total of 4938 subjects (2891 men and 2047 women with a mean age of 50.1 +/- 12.6 years), who had attended health examination at this hospital were enrolled. The Adult Treatment Panel III (ATP III) definition of MS components was adopted in this study with the exception of the definition of obesity. This was defined as body mass index (BMI) greater than 27 kg/m(2). Overall, 14% had high serum total triglyceride (TG), 8% had low high-density lipoprotein (HDL) cholesterol, and 18% were obese. WBC counts showed a statistically significant (P < .001) correlation with TG (r = .265), HDL(r = -.187), fasting glucose (r = .084), and BMI (r = .172) but not with blood pressure levels. In addition, RBC counts correlated significantly (P < .001) with TG (r = .250), HDL(r = -.269), fasting glucose (r = .098), and BMI (r = .228). WBC and RBC counts in subjects grouped according to the presence of 0, 1, 2, and >or= 3 features of MS were 6268 +/- 1633, 6555 +/- 1782, 6995 +/- 1880, and 7185 +/- 1696 cells/mm(3), and 4.63 +/ -0.56 x 10(6), 4.73 +/- 0.54 x 10(6), 4.84 +/- 0.60 x 10(6), and 4.91 +/- 0.55 x 10(6) cells/mm(3), respectively (P for trend <.001). Subjects in the highest quartile of WBC or RBC counts demonstrated a three- or twofold increase, respectively, in the odds ratio for MS with 3 or more metabolic features compared to subjects in the lowest quartile of WBC or RBC counts. Increased WBC and RBC counts, albeit normal, were associated with a variety of MS features in a Taiwan Chinese population, suggesting that hematological parameters could potentially be used as indicators of this syndrome. PMID- 15531182 TI - Ambulatory blood pressure in type 2 diabetic patients with albuminuria: relation to the renal function and structural lesions. AB - BACKGROUND/AIMS: To investigate possible relationships between ambulatory blood pressure (BP) and renal structure and function in type 2 diabetic patients. METHODS: Renal biopsies were performed on 39 patients with urine albumin concentrations above 100 mg/l. BP was investigated with a 24-h, automated, portable BP device. RESULTS: None of the patients in the study had signs of other renal disease than nephrosclerosis or diabetic nephropathy. Ten patients had slight, 13 intermediate, and 6 severe diabetic nephropathy on the renal biopsy. Among the remaining patients, 4 had normal microscopy findings and 6 had nephrosclerosis. The degree of albuminuria correlated to the systolic BP during the day (r = .43; P < .01) and night (r = .49; P < .01). The glomerular filtration rate (GFR) was associated with the systolic BP daytime (r = -.32; P < .05) and nighttime (r = -.47; P < .01). Neither degree of albuminuria nor GFR was associated with the diastolic BP levels. The degree of the glomerular pathology correlated to the systolic BP during daytime (P < .05), whereas the degree of interstitial fibrosis did not correlate to the BP levels. CONCLUSIONS: We have demonstrated that degree of albuminuria and GFR was significantly associated with daytime and nocturnal BP and glomerular structure with daytime BP. Furthermore, no renal disease other than diabetic nephropathy was found. PMID- 15531183 TI - Erythrocytic sorbitol contents in diabetic patients correlate with blood aldose reductase protein contents and plasma glucose levels, and are normalized by the potent aldose reductase inhibitor fidarestat (SNK-860). AB - The accumulation of sorbitol by the activated polyol pathway is considered to be a major cause of diabetic neuropathy. Because the erythrocytic sorbitol contents reportedly reflects that in nerves, erythrocytic sorbitol measurement would be useful for confirming the effect of an aldose reductase inhibitor (ARI). In this study, we examined erythrocytic sorbitol contents in healthy subjects and diabetic patients under fasting and postprandial conditions. Then, the contributions of blood aldose reductase (AR) contents and plasma glucose levels to the accumulated erythrocytic sorbitol contents were also analyzed. Erythrocytic sorbitol contents in the healthy subjects were 11.7 and 12.5-12.6 nmol/g Hb in fasting and postprandial status, respectively. In contrast, the erythrocytic sorbitol contents in diabetic patients were apparently higher (approximately 2.5-fold), but fidarestat treatment restored the elevated erythrocytic sorbitol contents to normal. In the diabetic patients, erythrocytic sorbitol contents were highly correlated with blood AR contents multiplied by the plasma glucose levels, whereas in the normal and fidarestat-treated diabetic patients no such correlation was observed. Taken together, these results suggest both the blood AR contents and the plasma glucose levels are factors determining erythrocytic sorbitol contents in diabetic patients. Notably, the potent ARI fidarestat was shown to normalize elevated erythrocytic sorbitol contents. PMID- 15531184 TI - Proinflammatory cytokines are increased in type 2 diabetic women with cardiovascular disease. AB - Diabetics have a much greater morbidity and mortality due to cardiovascular disease (CVD) than nondiabetics. Furthermore, diabetic women have a 3.8-fold greater risk for CVD compared to diabetic men. Inflammation is now considered a risk factor for CVD and it has been demonstrated that inflammation also plays a role in diabetes. One component of inflammation that has reported to be increased in patients with diabetes only and CVD only are proinflammatory cytokines, particularly interleukin-6 (IL-6), tumor necrosis factor (TNF-alpha), and interleukin-1 (IL-1beta). This study was performed to test the hypothesis that these proinflammatory cytokines were increased in women with CVD and further increased in diabetic women with CVD compared to nondiabetic women with CVD and healthy age-matched controls. We found that IL-6 was increased in diabetic women with CVD compared to healthy age-matched controls (1.41 = 0.48 to 0.33 +/- 0.06 pg/ml, P < .05). IL-6 was also increased in diabetic women without CVD compared to healthy age-matched controls, but not significantly (0.96 +/- 0.27 to 0.33 +/- 0.06 pg/ml). We found that TNF-alpha was increased in diabetic women with and without CVD compared to healthy age-matched controls, but not significantly (4.53 +/- 1.38 to 3.93 +/ -0.53 to 2.33 +/- 0.89 pg/ml). IL-1beta was not significantly different among any of the four groups of women. These results indicate that both IL-6 and TNF-alpha are chronically increased in diabetic women with and without CVD compared to nondiabetic women. The additive concentration of cytokines in diabetes and CVD suggests a common inflammatory state in both diabetes and CVD. PMID- 15531185 TI - Early subclinical limited axial and large joint flexibility in type 1 diabetes mellitus adolescents. AB - OBJECTIVE: The objective of this study was to compare the axial and large joint mobility in adolescents with and without type 1 diabetes mellitus (DM1). PATIENT AND METHODS: To check this relationship, 72 DM1 adolescents aged 9-20 years were admitted into the trial and compared with 46 healthy control subjects aged 10-18 years. The youths were compared with regard to anthropometrics (age, proportion female/male, weight, height, and BMI) data. The years from DM1 diagnosis and HbA(1c) (index) were 4.9 +/- 3.6 years and 1.40 +/- 0.39%, respectively. The values of the tests of flexibility of the movements of cervical joint, the abduction of scapular, wrist and back-lumbar joints and abduction of lame-femoral were obtained through the Fleximeter. RESULTS: The DM1 patients and controls did not differ regarding age (DM1 median 16, range 9-20 years vs. controls 16, range 10-18 years) and BMI (DM1 mean+/-S.D. 21.49 +/- 3.69 kg/m(2) vs. controls 20.76 +/- 2.81 kg/m(2)). The scapular, back-lumbar, and lame-femoral flexibility were, respectively, significantly lower (P < .001) in DM1 adolescents (175 +/- 8 degrees , 107 +/- 4 degrees , 66 +/- 10 degrees) compared with controls (189 +/- 13, 116 +/- 14, 76 +/- 12), but the cervical joint mobility was the same in both groups (DM1: 98 +/- l2 degrees vs. control: 101 +/- 13 degrees). CONCLUSION: Thus, the results of our study show a subclinical limited axial and large joint mobility in DM1 adolescents. Future prospective studies are needed to ascertain whether the joint limitations found in these DM1 adolescents will persist into adulthood and play a role in the development of other diabetic complications. PMID- 15531186 TI - Effects of insulin on the acetylcholine-induced hyperpolarization in the guinea pig mesenteric arterioles. AB - BACKGROUND: Insulin induces endothelium-dependent vasodilatation, which may be casually related to the insulin resistance and hypertension. Endothelium-derived nitric oxide (NO) is the most important mechanism of insulin-induced vasodilatation, and a possible contribution of endothelium-derived hyperpolarizing factor (EDHF) is also considered. Attempts were made to observe the effects of insulin on acetylcholine (ACh)-induced hyperpolarization in the submucosal arteriole of the guinea pig ileum, the objective being to investigate possible involvement of EDHF in the actions of insulin. METHODS: Conventional microelectrode techniques were applied to measure the membrane potential of smooth muscle cells in the submucosal arteriole. EDHF-induced hyperpolarization was elicited by ACh in the presence of both N(omega)-nitro-L-arginine (L-NNA) (100 microM) and diclofenac (1 microM). RESULTS: The resting membrane potential was -70.9 mV, and Ba(2+) (0.5 mM) depolarized the membrane to -33.0 mV. Insulin (10 microU/ml to 100 mU/ml) did not change the membrane potential in the absence or presence of Ba(2+). In the presence of Ba(2+), ACh (3 microM) hyperpolarized the membrane with two components, an initial large hyperpolarization followed by a slow and small one. Low concentration of insulin (100 microU/ml) did not alter the ACh-induced hyperpolarization. High concentration of insulin (100 mU/ml) shortened the time required to reach the peak amplitude and tended to increase the peak amplitude of the ACh-induced hyperpolarization. CONCLUSIONS: The data show that insulin enhances the ACh-induced hyperpolarization in the submucosal arterioles of the guinea pig ileum. The results suggested that EDHF also accounts for one of the endothelial factors involved in the insulin-induced vasodilatation. PMID- 15531187 TI - Breast arterial calcifications associated with diabetes and hypertension. AB - AIM: To investigate the relationship between breast arterial calcifications (BACs) and systemic hypertension (HT) and diabetes mellitus (DM). METHODS: Mammograms and patient records of 2406 women who were screened for breast cancer or had undergone diagnostic mammography were reviewed retrospectively. Mammograms were evaluated for the presence of arterial calcification and results were coded. Patients who had been using insulin or oral hypoglycemic agents were included in the diabetic group; patients who had been using antihypertensive agents were included in the hypertensive group. Diabetes was defined as use of oral hypoglycemic agents or insulin and hypertension was defined as use of antihypertensive agents. RESULTS: The prevalence of BACs among diabetics (25.4) was higher than among hypertensives (17.6%). The prevalence in the nondiabetic, nonhypertensive (NDNH) group was lowest (7.3%). The prevalence increased with age. BAC was seen almost four times more in diabetic patients and three times more in hypertensive patients than in NDNH controls. CONCLUSION: BACs are associated with diabetes and hypertension. BAC on a mammogram may indicate unsuspected diabetes or hypertension, especially after 59 years of age. PMID- 15531188 TI - The role of glimepiride in the effective management of Type 2 diabetes. AB - Type 2 diabetes mellitus, a disorder of impaired insulin secretion and insulin resistance, has reached epidemic proportions. The effective management of Type 2 diabetes is of vital concern to clinicians. The identification of high-risk individuals and lifestyle management can help control diabetes; however, most patients require pharmacologic intervention. The goals of pharmacologic therapy are to achieve adequate glycemic control while avoiding hypoglycemia and weight gain and to minimize the risk of future micro- and macrovascular complications. There are a number of available glucose-lowering agents from which to choose. This review focuses on the sulfonylureas, the first oral agents introduced for the management of Type 2 diabetes, which are effective, well-tolerated, and well established drugs, Second-generation sulfonylureas are now widely used in the management of Type 2 diabetes. The most recent addition, glimepiride, can be used in combination with metformin, the thiazolidinediones, alpha-glucosidase inhibitors, and insulin. The unique properties of glimepiride may provide advantages over other currently available insulin secretagogues. PMID- 15531189 TI - Ethical dilemmas regarding treatment when recruitment ends in randomized trials. PMID- 15531190 TI - Manipulation of the renin angiotensin system in peripheral arterial disease. AB - The Heart Outcomes Prevention Evaluation (HOPE) study has provided evidence for the use of ramipril for secondary cardiac prevention for patients with peripheral arterial disease. Despite this many vascular surgeons and general practitioners are reluctant to prescribe ACE inhibitors in a group of patients perceived to have a high incidence of renal artery stenosis. This review aims to review the pathophysiology of the renin-angiotensin system and make evidence based recommendations for commencing ACE inhibitors as part of a comprehensive delivery of best medical therapy to patients with peripheral arterial disease. PMID- 15531191 TI - Radiocephalic wrist arteriovenous fistula for hemodialysis: meta-analysis indicates a high primary failure rate. AB - OBJECTIVE: To improve the precision of the estimates of primary failure rates and primary and secondary 1 year patency of radial-cephalic arteriovenous fistulas (RCAVF) for hemodialysis. DESIGN: Meta-analysis. MATERIALS AND METHODS: A Medline search was performed of the English language medical literature between January 1970 and October 2002. Key words that were searched included radiocephalic fistula, arteriovenous shunt, Brescia-Cimino fistula and patency. Primary failure, primary and secondary patency rates were analysed using the standard mixed effects model, which allows for variability between the different studies. RESULTS: Eight prospective and 30 retrospective studies were included. The analysis showed a pooled estimated primary failure rate of 15.3% (95% CI: 12.7 18.3%). In addition, the pooled estimated primary and secondary patency rates of 62.5% (95% CI: 54.0-70.3%) and 66.0% (95% CI: 58.2-73.0%), respectively, were calculated. Subgroup analysis concerning various study characteristics, including study year, gender and age, did not reveal statistically significant differences. CONCLUSION: Although, the autogenous RCAVF is considered to be the primary choice for vascular access, this meta-analysis indicates a high primary failure rate and only moderate patency rates at 1 year of follow-up. PMID- 15531193 TI - Sapheno-femoral junction reflux in patients with a normal saphenous trunk. AB - OBJECTIVE: To determine the patterns and clinical importance of saphenofemoral junction (SFJ) reflux in patients with chronic venous disease (CVD) and a normal great saphenous vein (GSV) trunk. METHODS: Fifteen hundred consecutive patients were examined using duplex ultrasound (DU) in three centres. Patients with reflux involving the SFJ and/or its tributaries only were included and its prevalence and patterns were studied. Patients with GSV trunk reflux or in any other veins were excluded. The SFJ diameter was categorised as normal, dilated or varicose. The results of surgery were evaluated by DU in 42 patients 1 year after the procedure. RESULTS: SFJ area incompetence with a competent GSV trunk occurred in 8.8% of limbs. It was significantly more common in CEAP class 2, 13.6% compared to class 3, 8.2% (p=0.03), class 1, 2.7%, class 4, 4.4% and classes 5 and 6 together, 1.5% (p<0.001 for all). The SFJ had a normal diameter in 21%, dilated in 62% and varicose in 17%. Reflux was seen in 39% of limbs with a normal SFJ diameter, in 85% of those with a dilated SFJ and in all varicose SFJs. Of the 42 operated limbs, 27 had ligation and division of the SFJ and tributary phlebectomies. Fifteen had tributary phlebectomies only, leaving the SFJ intact. At one-year follow-up, SFJ area reflux was found in six limbs (14.3%), involving the SFJ alone in 1, a main tributary in 1 and 4 small tributaries. No reflux was found in the GSV trunk. All but two of the 42 patients were satisfied with the results. CONCLUSIONS: SFJ reflux with tributary involvement and sparing of the GSV trunk occurs in 8.8% of CVD patients. Such reflux is found in the entire spectrum of CVD, but it is more common in class 2. Local surgery with or without SFJ ligation has very good results at 1 year. DU scanning prior to treatment is important in all patients so that the intact GSV can be spared. PMID- 15531194 TI - Smooth muscle cell apoptosis in primary varicose veins. AB - OBJECTIVES: One of the important factors responsible for vessel wall remodelling is programmed cell death. In the paper the role of smooth muscle cell (SMC) apoptosis in primary varicose veins (PVV) is investigated. MATERIAL AND METHODS: Vein specimens were obtained from 40 patients with PVV. In each case proximal and distal (upper crural) great saphenous veins (GSV) were harvested. Morphometric computer assessed quantitative evaluation of SMCs, collagen and elastin content was carried out. Apoptotic cells were detected by TUNEL assay. The levels of p53, BAX, BCLl-2 and p21 mRNA expression were assessed by real time RT-QPCR and the presence of respective proteins in the vessel wall was confirmed by immunohistochemistry. RESULTS: In the proximal GSV segments a significant increase of p53, p21 and BCL-2 mRNA levels was found in PVV patients. In the distal segments BAX and BCL-2 expression levels were higher. Taking into account the patient age, elevated p53 mRNA expression level was noticed in the distal incompetent GSVs of young PVV patients. In this group a statistically significant increase in the apoptotic index (APIx) within the vein media was found which correlated positively with p53 mRNA expression level. There was no increase of the apoptotic activity in elderly patients that led to the structural changes increase. In proximal GSV segments, despite SMC amount reduction or presence of structural changes in perivalvular wall region, no increase of the APIx with was noticed. CONCLUSIONS: P53-related apoptosis is one of the regulatory mechanisms of vein wall homeostasis maintenance. During varicose vein development its activation is related to the early stages of the disease. In the further course, the down-regulation of the SMC apoptosis within the vein media leads to the structural changes increase. The reduction of the SMC population corresponding to an increase of p21 expression in proximal saphenous vein segments suggests that the cell cycle disturbances may lead to the 'weakness' of the proximal GSV wall. Valve injury is not the only factor leading to the varicose veins occurrence. PMID- 15531192 TI - Saphenofemoral venous channels associated with recurrent varicose veins are not neovascular. AB - BACKGROUND: Recurrence of varicose veins after apparently adequate surgery is common. Neovascularisation, the formation of new vascular channels between a venous surgery site and new varicosities, is thought to be an important cause of recurrence. The aim of this study was to provide histological evidence of the 'neovascularisation'process. METHOD: Tissue samples from the region of the previously ligated saphenofemoral junction (SFJ) were taken from 14 limbs with recurrent varicose veins and from nine control limbs. Tissue samples were analysed histologically for overall vascularity, and the presence of intimal circular fibrosis, intimal eccentric fibrosis, medial thickened elastosis, and thrombosis in the microscopic thin walled vessels within the tissue. The same samples were analysed immunohistoligically for S100, a neural marker, and Ki-67 (Mib 1), a marker of endothelial proliferation. Absent S100 and positive Ki-67 were considered as evidence of new vessels. RESULT: No significant difference was found between the venous recurrence and control groups in respect to histological features. S100 positive nerve fibrils were seen associated with dilated venous channels in the majority of both redo and control groups (p=1, Fisher's exact test). Only one section stained positively with Ki-67 (Mib1) in a single vascular channel for a few endothelial cells. The remaining control and redo cases were negative for Mib 1 (p=1, Fisher's exact test). CONCLUSION: We found little evidence of neovascularisation associated with recurrent varicose veins in the saphenofemoral region. The venous channels that develop at the previously ligated SFJ may represent adaptive dilatation of pre-existing venous channels (vascular remodelling), probably in response to abnormal haemodynamic forces. PMID- 15531195 TI - Emergency non-ruptured abdominal aortic aneurysm. AB - OBJECTIVES: To investigate symptoms and early mortality (<30 days) following open surgery for emergency, symptomatic non-ruptured abdominal aortic aneurysm (AAA). DESIGN: Retrospective cohort study. PATIENTS AND METHODS: During the period 1983 1994, 129 patients had an emergency admission, followed by surgery, for symptomatic non-ruptured AAA. Sixty-one received surgery within 24 h of admission and 68 received surgery more than 24 h after admission (median 135 h, inter quartile range: 51-239 h). During the same period 239 patients had elective surgery for non-ruptured AAA. Early mortality (<30 days), symptoms and co morbidities were recorded. Data were retrieved from the patient records. RESULTS: Mortality (30 days) was 18% in the 61 patients having surgery within 24 h of emergency admission for non-ruptured AAA. Mortality following either delayed surgery (semi-elective) after emergency admission or elective surgery was 4.2% (p=0.0002). Four out of 11 patients who died within 30 days following an acute operation had previously been declared unfit for elective surgery. One additional emergency patient had been found unfit for open surgery, but survived a delayed operation. CONCLUSION: The high mortality rate of patients with non-ruptured, symptomatic AAA undergoing surgery within 24 h of admission appears to be influenced by several factors, including co-morbidities and the acute operation. We propose that the 30-day mortality for non-ruptured AAA should be reported in two categories: mortality rate for elective surgery and mortality for surgery performed within 24 h of emergency admission. The term 'emergency non-ruptured' is a suitable term for the latter group. PMID- 15531196 TI - Total laparoscopic aortic surgery: transperitoneal left retrorenal approach. PMID- 15531197 TI - Pulsatile wall motion (PWM) measurements after endovascular abdominal aortic aneurysm exclusion are not useful in the classification of endoleak. AB - The pulsatile wall motion (PWM) of AAA is reduced after endovascular stent-graft placement. The purpose of this study was to identify whether PWM after endografting was useful in the classification of endoleak. PATIENTS AND METHODS: 162 patients treated with EVAR underwent pre- and post-operative PWM assessment with ultrasonography. Follow-up was 1-9 years. 111 patients had well-excluded aneurysms, three patients had enlarging aneurysms without any recognizable endoleak (endotension), 16 had type I, 31 had type II and 1 had type III endoleak. RESULTS: The PWM was reduced from about 1mm pre-operatively to 0.24 mm post-operatively in well-excluded aneurysms. PWM remained stable during follow up. Type I endoleak was associated with moderately reduced PWM (proximal endoleak 0.79 mm and distal 0.32 mm). PWM in patients with type II endoleak was higher (0.32 mm) post-operatively (p=0.002) compared to well-excluded aneurysms. CONCLUSION: PWM is permanently reduced after endografting. The smallest reduction in PWM was in patients with type II endoleaks. However, the overlap between the groups does not allow reliable identification of patients having endoleak with PWM-measurements. PMID- 15531198 TI - First clinical experiences with an endovascular clamping system for neuroprotection during carotid stenting. AB - OBJECTIVES: This report evaluates the feasibility and efficacy of an endovascular blood flow blockage technique to prevent intracerebral embolization of plaque debris during carotid artery stenting. METHODS: Forty-two patients were enrolled in five clinical sites in Germany and Italy with either an asymptomatic internal carotid artery stenosis > or =75% (mean 87%) or a symptomatic stenosis > or =60% (mean 85%). Cerebral protection during the stenting procedure was achieved using an endovascular clamping technique, obtained by occlusion of the external and common carotid artery via two independently inflatable balloons integrated in the Mo.Ma system. Blood with particulate plaque debris was aspirated before flow was restored. The patient's clinical and the neurological status were assessed during intervention, at discharge, and at 3 months follow-up. RESULTS: Stenting was performed in all but one patient. The mean flow occlusion time was 10.6+/-6.5 min. Transient clamping intolerance was observed in five patients (12%). In two patients, neurological deficits persisted for 2 and 12 h, respectively. Two minor strokes (4,7%) occurred at 5 and 72 h after the procedure. No major strokes or deaths were observed at 3 months follow-up. CONCLUSIONS: This first clinical experience with the Mo.Ma device substantiates the feasibility of endovascular clamping in preventing cerebral embolization during carotid artery stenting. PMID- 15531199 TI - Preoperative duplex on admission prevents unnecessary carotid surgery. AB - OBJECTIVES: Our unit policy is to duplex on admission all patients undergoing carotid endarterectomy to confirm patency. The aim of this study was to evaluate whether this had led to avoidance of unnecessary carotid surgery in a significant number of patients. METHODS: Over a 7-year period from January 1997 to December 2003, a total of 475 patients were scheduled for carotid endarterectomy. Of these, 439 patients subsequently underwent carotid endarterectomy. These data were obtained from prospectively collected vascular departmental records and we also hand searched notes of the 36 cancelled patients. RESULTS: There were a total of 36 cancellations for various reasons. Of these, 8 were clearly occluded on duplex. A further 16 were highly suspicious of occlusion on duplex and thus surgery was deferred and selective angiography was undertaken. Of these 9 were confirmed to be occluded on angiogram and a further 4 had severe stenoses or virtual occlusion, 3 patients had a distal internal carotid artery occlusion. Thus a total of 24 patients had an occlusion either diagnosed directly on duplex or because of suspicion on duplex. Unnecessary carotid surgery was avoided in 24 of 475 (5%) of patients. CONCLUSION: This study shows that a preoperative duplex on admission for surgery results in 1 of 20 patients avoiding unnecessary carotid surgery. PMID- 15531200 TI - Prolongation of the intestinal viability using oxygenated perfluorocarbon in an experimental model of acute intestinal ischemia. AB - OBJECTIVES: Liquid perfluorocarbons (PFCs) are well known for their capability to carry respiratory gases. The aim of this study was to evaluate the effectiveness of oxygenated F-Decalin on the intestinal viability, in an experimental model of acute intestinal ischemia. MATERIAL AND METHODS: Thirty-six rabbits were subjected to 8h intestinal ischemia by ligation of the superior mesenteric artery (subgroups 1), the mesenteric vein (subgroups 2) or both vessels (subgroups 3). The animals were divided into three groups: (a) Control (ischemia alone), (b) PFC O2 (ischemia plus infusion of oxygenated F-Decalin) and (c) PFC (ischemia plus infusion of not-oxygenated F-Decalin). Intestinal biopsies from four different sites and blood samples for serum enzymes measurements were taken at 2, 4, 6 and 8 h. All tissue sections were examined blindly under light microscope. Sections from the specimens were taken at 4 and 8 h, and examined blindly under the electron microscope. Statistical analysis was performed by non-parametric Kruskal Wallis test. RESULTS: Using light microscope, the observed intestinal damages to the sections from Control and PFC groups were severe at 4 h and destructive after 8 h. On the contrary, minimal injuries were observed in the biopsies from PFC-O2 group at 4 and even after 8 h of ischemia. These findings were confirmed by the electron microscope study and correlated to the serum enzymes measurements. CONCLUSIONS: These results suggest that intestinal viability could be prolonged after acute ischemia using oxygenated perfluorocarbons and this could be a promising pretreatment modality for a variety of mesenteric ischemic forms. PMID- 15531201 TI - Sites of implantation for central venous access devices (ports): a study of the experiences and preferences of patients. AB - OBJECTIVE: To obtain information which might guide vascular specialists and their patients in the choice of site for implantation of central venous access devices (CVADs). DESIGN: Questionnaire study. METHODS: Questionnaires were sent to 69 patients with cystic fibrosis and 54 (78%) responded (39 females: age 5-63, median 24 years). They had received a total of 79 CVADs placed in the upper chest (60), lower chest (13), thigh (3) and arm (3). Only 46% patients had been offered a choice of site. RESULTS: Questions about 14 specific areas of disability or concern found problems most frequently with discomfort (54%), wearing a seatbelt (51%), cosmetic appearance (44%), scarring (44%), choice of clothing (42%) and lying in bed or sleeping (42%). There were no significant differences between upper and lower chest CVADs. Patients with upper chest CVADs seldom had any problems with use of their arm (12%). 81% CVADs could not be accessed by the patients, and in 39% of these cases patients would have liked to do so. CONCLUSIONS: Many patients complain of few problems with their CVADs, regardless of site, but half have some persistent discomfort. Cosmetic considerations frequently cause concern and patients should be given choice in the site of their CVADs. PMID- 15531202 TI - Subintimal angioplasty in the treatment of patients with intermittent claudication: long term results. AB - OBJECTIVES: Reporting the long-term results of subintimal angioplasty (SA) in patients with intermittent claudication (IC). DESIGN: A prospective study. PATIENTS: One hundred and sixteen SA procedures were performed in 104 patients, from February 1997 to January 2000. METHODS: This is a prospective study of patients treated for IC with infrainguinal SA. Primary assisted patency rates were calculated, also on intention to treat basis. Univariate and multivariate Cox regression tests were used to assess whether patency was correlated with co morbidities, run-off or occlusion length. RESULTS: There was no early mortality. Technical success was achieved in 101 cases (87%). Primary assisted patency rates on intention to treat basis (116 cases) at 6, 12, 36 and 60 months were 69, 62, 57 and 54%, respectively. For successfully recanalized patients (101 cases) these respective numbers are 79, 70, 66 and 64%. Length of occlusion, age and male gender were independent risk factors for reocclusion. CONCLUSIONS: The satisfactory results obtained in the present study are probably due to two main factors. First, the three participating radiologist are highly skilled and experienced. Secondly, a conscientious surveillance was adhered to, so that restenoses could be diagnosed and treated early. SA is a relevant alternative to bypass surgery in patients with disabling IC due to long femoro-popliteal occlusions. It is far less traumatic than conventional vascular reconstructions, complications are few and not serious. Very importantly, SA never interfered with later successful vascular surgery. Therefore, we have adopted SA as the primary treatment for patients with IC when medical treatment alone has not been satisfactory. PMID- 15531203 TI - Modulation of human adventitial fibroblast function by photodynamic therapy of collagen matrix. AB - OBJECTIVES: Photodynamic therapy (PDT) is a promising strategy to limit restenosis. PDT depletes the resident cells from the vessel wall without adventitial cell ingrowth. This study was undertaken to further explore the mechanisms by which PDT of matrix acts on key mechanisms in the development of restenosis. MATERIALS AND METHODS: Control and PDT-treated collagen type-I matrix gels were prepared. Thereafter, untreated human fibroblasts were seeded on matrix gels (n=12). Fibroblast proliferation and invasive migration were quantified by calibrated phase contrast microscopy. Fibroblast bFGF and TGF-beta1 mRNA expression were analyzed using a quantitative real-time reverse transcription polymerase chain reaction. RESULTS: Fibroblast proliferation on PDT-treated matrix gels was reduced by 30 and 76% after 3 and 7 days, respectively (3 days: P or =6 mice) cut in the direction of longitudinal muscle were studied. Numerous conditions of electrical field stimulation (EFS) and effects of exogenous NO and NO donors were studied in the absence or presence of inhibitors of nitric oxide synthase (NOS) and 1H-[1,2,4]-oxadiazaolo-[4,3-a]-quinoxalin-1-one (ODQ), a downstream inhibitor of guanylyl cyclase. EFS induced a frequency-dependent inhibition of contractile activity in both jejunum and ileum (P < 0.05). As the voltage of EFS was increased, inhibition turned to excitation in the jejunum; in contrast, the ileum demonstrated a voltage-dependent increasing inhibition (P < 0.05 each). EFS induced inhibition was blocked by NOS inhibitors and ODQ. NO donors inhibited spontaneous contractile activity abolished by ODQ. NO appears to be an endogenous inhibitory neurotransmitter in murine longitudinal small bowel muscle. Nitrergic mechanisms mediate inhibitory control of murine longitudinal small intestinal muscle. Differences exist in neuroregulatory control between jejunum and ileum that may be related to their known difference in motor patterns. PMID- 15531237 TI - Inhibition of the vanilloid receptor subtype-1 attenuates TNBS-colitis. AB - Primary sensory neurons are important in regard to the initiation and propagation of intestinal inflammation. The vanilloid receptor subtype-1 (VR-1) is a cation channel located on the sensory nerves that, when stimulated, release proinflammatory peptides. Previous reports have indicated that inhibition of VR-1 with capsazepine (CPZ), a VR-1 antagonist, attenuates dextran sodium sulfate (DSS) colitis in rats. DSS-induced colitis resembles ulcerative colitis with regard to its pathologic features. In this study, we examined the effect of CPZ on trinitrobenzene sulfonic acid (TNBS)-induced colitis, an experimental model of intestinal inflammation that most closely resembles the histologic and microscopic features of Crohn's disease. Colitis was induced by administering a single enema of 100 mg/kg TNBS in 50% ethanol via catheter to lightly anesthetized rats. Subsets of rats were treated with either 1 micromol/kg/ml of CPZ or CPZ-vehicle via enema for 6 days. Seven days after TNBS administration, rats were sacrificed and inflammation was assessed using a validated macroscopic damage score (MDS) and by measuring myeloperoxidase (MPO) activity. In addition, histologic examination was performed. TNBS administration resulted in reproducible chronic erosive lesions extending into the muscularis propria and extensive recruitment of neutrophils in the distal colon. MDS and MPO scores were considerably elevated in the TNBS colons when compared with the TNBS vehicle animals. TNBS rats treated with CPZ enemas exhibited a substantial reduction in MDS and MPO scores and demonstrated dramatically improved pathologic findings. Topical CPZ resulted in considerable attenuation of TNBS-induced colitis. These results support the role of VR-1 and sensory neurons with regard to intestinal inflammation. PMID- 15531238 TI - Effect of body mass index on nonalcoholic fatty liver disease in patients undergoing minimally invasive bariatric surgery. AB - The risk factors for nonalcoholic fatty liver disease in patients undergoing bariatric surgery are under study. We wanted to determine the correlation between nonalcoholic fatty liver disease and patient factors such as obesity and liver function tests. A retrospective analysis was performed on 177 nonalcoholic morbidly obese patients who underwent laparoscopic Roux-en-Y gastric bypass with liver biopsy, to identify risk factors for nonalcoholic fatty liver disease. The histologic grade of liver disease was compared with preoperative body mass index, age, and liver function tests. Simple steatosis and steatohepatitis were present in 90% and 42% of patients, respectively. Elevated transaminase levels were an independent risk for liver disease. Body mass index and liver disease were not correlated with univariate analysis. Regression analysis performed on age, body mass index, and liver disease demonstrated that the risk for liver disease increased with body mass index in the younger (<35 years old) age group and decreased with body mass index in the older (>45 years old) age group. There was a high incidence of steatosis and steatohepatitis in these nonalcoholic bariatric patients, and elevated transaminase level was indicative of disease. Body mass index was a positive risk factor for liver disease in younger patients but a negative risk factor in the older patients. PMID- 15531239 TI - The practice of bariatric surgery at academic medical centers. AB - The growing demand for laparoscopic bariatric surgery has led to an increase in the development of new bariatric surgical practices. Proper hospital facilities and an experienced bariatric surgical team are necessary to ensure optimal patient results. We surveyed academic centers participating in the University HealthSystem Consortium to examine the current practice of bariatric surgery. The survey questioned (1) availability of resources and equipment designed for the morbidly obese, (2) accidents, equipment problems, and workers' compensation relating to the care of bariatric surgical patients, (3) credentialing of bariatric surgeons, and (4) suggestions for improvements in the bariatric surgery program. Twenty-five institutions that perform bariatric surgery responded. Although the majority of institutions noted that they had basic bariatric equipment, some organizations did not have facility resources such as high-weight operating room tables and computed tomography scanners or transfer devices. Twenty-eight percent of institutions reported having accidents or equipment problems and 40% of institutions had workers' compensation claims relating to the care of bariatric patients. With regard to credentialing, 60% of institutions required the surgeons to have performed a minimum number of procedures prior to granting privileges. Suggested improvements included the need for more specialized bariatric equipment, enhancement of the education of all members of the bariatric surgical team, and designation of a bariatric physician who would coordinate care. This survey of bariatric surgery practices at academic medical centers demonstrates that the practice of bariatric surgery could be improved with regard to availability of bariatric equipment and resources and credentialing of surgeons. PMID- 15531241 TI - The benefits of a dedicated minimally invasive surgery program to academic general surgery practice. AB - In 2001, a dedicated minimally invasive surgery (MIS) program was established at a large university hospital. Changes included improvement and standardization of equipment and instruments, patient care protocols, standardized orders, and staff education. The aim of this study was to evaluate the impact of this program on an academic surgery practice. From January 1999 through October 2003, hospital and departmental databases were reviewed for all records pertaining to general surgery cases. Data trends were analyzed by regression analysis and are expressed as mean +/- SEM. In 1999, 15.0 +/- 0.1% of all general surgery cases were MIS cases compared with 30.2 +/- 0.1% in 2003 (P < 0.0001). During this period, the number of patients requiring conversion from a laparoscopic to an open approach decreased from 14.4% to 4.0% (P = 0.0007). In 1999, 30% of appendectomies were laparoscopic, compared with 92% in 2003 (P < 0.0001). This increase in the rate of laparoscopic appendectomy resulted in a decrease in average length of hospital stay for all patients with acute appendicitis, from 5.5 +/- 1.0 days in 1999 to 2.7 +/- 0.2 days in 2003 (P < 0.0001), and a decrease in total hospital cost per case, from 6569 +/- 400 US dollars in 1999 to 4819 +/- 175 US dollars in 2002 (P < 0.001). Total operating room time per case for cholecystectomy decreased from 131 +/- 3.7 to 108 +/- 3.2 minutes (P < 0.0001), and actual surgery time decreased from 95 +/- 4.1 to 74 +/- 4.0 minutes (P = 0.0006). Implementation of a dedicated MIS program resulted in a significant increase in the number of MIS cases and percentage of general surgery cases performed by MIS. This increase in the utilization of MIS resulted in reduced length of stay and cost and has been accompanied by improvements in operating room efficiency. Changes in practice associated with development of an MIS program have had measurable institutional benefits. PMID- 15531240 TI - G protein polymorphisms do not predict weight loss and improvement of hypertension in severely obese patients. AB - Both the gene encoding the alpha subunit of G stimulatory proteins (GNAS1) and the beta3 subunit gene (GNB3) of G proteins are associated with obesity and/or hypertension. Moreover, the TT/TC825 polymorphism of GNB3 predicts greater weight loss than the CC825 polymorphism in obese patients (mean body mass index, 35 kg/m2) undergoing a structured nonpharmacologic weight loss program. Gastric banding enforces a low-calorie diet by diminishing the need for volitional adherence. It is unknown whether these polymorphisms predict the variable weight loss in patients after bariatric surgery. Three hundred and four severely obese patients (mean +/- SEM age, 42 +/- 1 years; 245 women and 59 men; mean +/- SEM body mass index, 43.9 +/- 0.3 kg/m2) followed prospectively for at least 3 years after surgery were genotyped for the GNB3 C825T, G814A, and GNAS1 T393 polymorphisms. All analyses were performed blinded to the phenotypic characteristics of the study group. Frequencies of polymorphisms were comparable to those previously published. No polymorphism studied predicted 3-year weight loss or was associated with high blood pressure in severely obese patients after gastric banding. Multivariate analysis of potentially confounding factors such as reoperation rate or use of sibutramine or orlistat revealed similar results (P > 0.1). Regardless of the mechanism(s) involved for these discordant findings, GNB3 C825T, G814A, and GNAS1 T393C polymorphisms do not seem to be reliable predictors of long-term weight loss. PMID- 15531242 TI - Visuospatial skills and computer game experience influence the performance of virtual endoscopy. AB - Advanced medical simulators have been introduced to facilitate surgical and endoscopic training and thereby improve patient safety. Residents trained in the Procedicus Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR) laparoscopic simulator perform laparoscopic cholecystectomy safer and faster than a control group. Little has been reported regarding whether factors like gender, computer experience, and visuospatial tests can predict the performance with a medical simulator. Our aim was to investigate whether such factors influence the performance of simulated gastroscopy. Seventeen medical students were asked about computer gaming experiences. Before virtual endoscopy, they performed the visuospatial test PicCOr, which discriminates the ability of the tested person to create a three-dimensional image from a two-dimensional presentation. Each student performed one gastroscopy (level 1, case 1) in the GI Mentor II, Simbionix, and several variables related to performance were registered. Percentage of time spent with a clear view in the endoscope correlated well with the performance on the PicSOr test (r = 0.56, P < 0.001). Efficiency of screening also correlated with PicSOr (r = 0.23, P < 0.05). In students with computer gaming experience, the efficiency of screening increased (33.6% +/- 3.1% versus 22.6% +/- 2.8%, P < 0.05) and the duration of the examination decreased by 1.5 minutes (P < 0.05). A similar trend was seen in men compared with women. The visuospatial test PicSOr predicts the results with the endoscopic simulator GI Mentor II. Two-dimensional image experience, as in computer games, also seems to affect the outcome. PMID- 15531243 TI - Evaluation of vagus nerve function before and after antireflux surgery. AB - We sought to evaluate vagus nerve integrity before and after antireflux surgery and to compare it with symptomatic outcome. Antireflux surgery patients were recruited. Patients with disorders associated with vagus dysfunction or who took medications with anticholinergic effects were excluded. Each patient underwent a sham-feeding-stimulated pancreatic polypeptide (PP) test before and after surgery. A symptom survey was also administered. Twenty patients completed preoperative testing; their mean age was 57 years, and postoperative testing results were available for 16 of them. Of the 20, 14 (70%) had an appropriate increase in PP level with sham-meal preoperatively. All 4 patients with an abnormal preoperative test remained abnormal, and 5 of 12 (42%) with a normal preoperative test had an abnormal postoperative result; thus 9 of 16 (56%) had an abnormal postoperative PP test. In 15 patients, assessments of bowel function were obtained before and after surgery. Six of 15 (40%) patients developed new or worse symptoms (diarrhea in 4, flatus in 2). The symptoms did not correlate with PP results. This suggests that some patients referred for antireflux surgery have evidence of abnormal vagus function that persists after surgery. Many patients (42%) with normal testing before surgery develop an abnormal test after surgery. There was no correlation between PP tests and the development or worsening of bowel symptoms. PMID- 15531244 TI - Omeprazole does not reduce gastroesophageal reflux: new insights using multichannel intraluminal impedance technology. AB - Proton pump inhibitors are the mainstay of medical management in gastroesophageal reflux disease. Although they provide relief from most symptoms, reflux may persist. We hypothesize that omeprazole does not reduce the total amount of gastroesophageal reflux but simply alters its pH characteristics. Six asymptomatic volunteers had combined 24-hour impedance pH monitoring before and after 7 days of omeprazole (20 mg BID). Multichannel intraluminal impedance was used to identify reflux episodes, which were classified as acid (pH < 4), weak acid (pH > 4 but decrease > 1 pH unit) and nonacid (pH > 4 and decrease < 1 pH unit) by pH measurements 5 cm above the lower esophageal sphincter (LES). A gastric pH sensor located 10 cm below the LES was used to verify the action of omeprazole. Impedance detected a total of 116 reflux episodes before and 96 episodes after omeprazole treatment. The median number of reflux episodes (18 versus 16, P = 0.4), median duration of reflux episodes (4.7 versus 3.6 minutes, P = 0.5), and total duration of reflux episodes (27.2 versus 42.4 minutes, P = 0.5) per subject were similar before and after omeprazole. Acid reflux episodes were reduced from 63% before to 2.1% after omeprazole (P < 0.0001), whereas nonacid reflux episodes increased (15% to 76%, P < 0.0001). Weak acid reflux episodes did not change (22.4% to 21.8%, P = 1.0). The proportion of reflux episodes greater than pH 4 increased from 37% to 98% (P < 0.0001). In normal subjects, omeprazole treatment does not affect the number of reflux episodes or their duration; rather it converts acid reflux to less acid reflux, thus exposing esophagus to altered gastric juice. These observations may explain the persistence of symptoms and emergence of mucosal injury white on proton pump inhibitor therapy. PMID- 15531245 TI - Multiorgan resection for gastric cancer: intraoperative and computed tomography assessment of locally advanced disease is inaccurate. AB - Multiorgan resection of locally advanced gastric cancer has previously been associated with increased morbidity. This study was performed to determine the actual prevalence of pathologic T4 disease in multiorgan gastric resection specimens excised for presumed clinical T4 gastric cancer. A prospective oncology database was queried to identify gastric cancer patients who underwent en bloc multiorgan resection for clinical T4 lesions. Four hundred eighteen patients with gastric cancer underwent gastrectomy between 1990 and 2002. Multiorgan resection was performed in 21 of 418 (5%) patients. Multiorgan resection was not associated with a significant increase in morbidity or mortality. Pathologically confirmed T4 disease was present in only 8 of 21 (38%) patients; the pathologic T stage in all remaining patients was T3 (13 [62%]). Fifteen patients were evaluated by preoperative computed tomography scan. Preoperative computed tomography was inaccurate in assessing T4 lesions, with a positive predictive value of only 50%. Multiorgan resection was safely performed in patients with locally advanced gastric cancer. Pathologic T4 disease was present in only one third of multiorgan resections performed for en bloc excision of locally advanced gastric cancer. Improved methods for intraoperative assessment of disease extension to adjacent viscera should be investigated. PMID- 15531247 TI - A rare diagnosis for a pancreatic mass: splenosis. AB - Splenosis, the autotransplantation of splenic tissue, has been designed to preserve organ functions after splenectomy. We present the first case of laparoscopic resection of a pancreatic splenosis, in a patient who had undergone a splenectomy 31 years before, complaining of abdominal pain and diarrhea. Abdominal computed tomography (CT) scan showed an enhancing hypervascular 3-cm solid mass in the body of the pancreas, mimicking a pancreatic cancer or a neuroendocrine tumor. A diagnostic laparoscopy was planned, and a 3-cm peripancreatic nodule with a long pedicle was visualized, with many nodules close to the tail of the pancreas and in the greater omentum. They were all resected, and the specimens obtained were immediately sent for frozen-section examination, which confirmed the diagnosis of heterotopic splenic tissue. Splenosis should be included in the differential diagnosis of the pancreatic masses in patients with previous splenic surgery. A hypervascular mass on CT scan should be regarded as an adenocarcinoma of the pancreas until proven otherwise. The possibility of a neuroendocrine tumor mandates an octreotide scan and gastrointestinal hormones dosage. In the unlikely event that all tests may produce equivocal results, a diagnostic laparoscopy is mandatory, in order to obtain an accurate histopathologic diagnosis. PMID- 15531248 TI - The SARS epidemic in 2002-2003 shocked the world. PMID- 15531246 TI - Current management of gastric cancer. PMID- 15531249 TI - The chronology of the 2002-2003 SARS mini pandemic. AB - Severe acute respiratory syndrome (SARS) was a new human disease in the autumn of 2002. It first occurred in Southern China in November 2002 and was transported to Hong Kong on February 21, 2003 by an infected and ill patient. Ten secondary cases spread the infection to two hospitals in Hong Kong and to Singapore, Toronto and Hanoi. In March 2003 a novel coronavirus (SARS-CoV) was found to be the causative agent. Within 11 weeks from the first SARS case in Hong Kong it had spread to an additional 27 countries or special administrative regions. The mini pandemic peaked during the last week of May 2003 and the last new probable case was on July 13, 2003. There were a total of 8096 probable cases and 774 deaths. Sixty-six per cent of the cases occurred in China, 22% in Hong Kong, 4% in Taiwan and 3% in both Singapore and Canada. Twenty-one per cent of all cases occurred in healthcare workers. PMID- 15531250 TI - Epidemiology of severe acute respiratory syndrome (SARS): adults and children. AB - Severe acute respiratory syndrome (SARS) is a newly described respiratory infection with pandemic potential. The causative agent is a new strain of coronavirus most likely originating from wild animals. This disease first emerged in November 2002 in Guangdong Province, China. Early in the outbreak the infection had been transmitted primarily via household contacts and healthcare settings. In late February 2003 the infection was transmitted to Hong Kong when an infected doctor from the mainland visited there. During his stay in Hong Kong at least 17 guests and visitors were infected at the hotel at which he stayed. By modern day air travel, the infection was rapidly spread to other countries including Vietnam, Singapore and Canada by these infected guests. With the implementation of effective control strategies including early isolation of suspected cases, strict infection control measures in the hospital setting, meticulous contact tracing and quarantine, the outbreak was finally brought under control by July 2003. In addition, there were another two events of SARS in China between the end of December 2003 and January 2004 and from March to May 2004; both were readily controlled without significant patient spread. PMID- 15531251 TI - Clinical picture, diagnosis, treatment and outcome of severe acute respiratory syndrome (SARS) in children. AB - Children are susceptible to infection by SARS-associated coronavirus (SARS-CoV) but the clinical picture of SARS is milder than in adults. Teenagers resemble adults in presentation and disease progression and may develop severe illness requiring intensive care and assisted ventilation. Fever, malaise, cough, coryza, chills or rigor, sputum production, headache, myalgia, leucopaenia, lymphopaenia, thrombocytopaenia, mildly prolonged activated partial thromboplastin times and elevated lactate dehydrogenase levels are common presenting features. Radiographic findings are non-specific but high-resolution computed tomography of the thorax in clinically suspected cases may be an early diagnostic aid when initial chest radiographs appear normal. The improved reverse transcription polymerase chain reaction (RT-PCR) assays are critical in the early diagnosis of SARS, with sensitivity approaching 80% in the first 3 days of illness when performed on nasopharyngeal aspirates, the preferred specimens. Absence of seroconversion to SARS-CoV beyond 28 days from disease onset generally excludes the diagnosis. The best treatment strategy for SARS among children remains to be determined. No case fatality has been reported in children and the short- to medium-term outcome appears to be good. The importance of continued monitoring for any long-term complications due to the disease or its empiric treatment, cannot be overemphasised. PMID- 15531252 TI - Post-SARS infection control in the hospital and clinic. AB - The recent severe acute respiratory syndrome (SARS) outbreak has almost mandated a re-evaluation of infection control practices in hospitals, clinics, schools and domestic environments, especially for patients with respiratory tract symptoms. Triage, early case detection followed by prompt isolation and quarantine are major preventive measures. Respiratory tract infections are the most common childhood illnesses and paediatric SARS poses special problems in diagnosis because of its non-specific presentation. The main lessons learnt from the outbreak were: (1) despite well established guidelines on infection control precautions, poor understanding of underlying principles and deficiencies in compliance are common among healthcare professionals, especially during emergencies; (2) even a slight lapse can be fatal; and (3) over-protection can be counterproductive. Hence it is important to: (1) be protected to protect others; (2) be vigilant and prepared for emerging infections; (3) be proficient and scrupulous in infection control measures; (4) be apposite and practical on personal protective equipments to ensure sustainability; and (5) be dutiful and prompt in informing of potential threats and work closely with others. PMID- 15531253 TI - Long-term sequelae of SARS in children. AB - Severe acute respiratory syndrome (SARS) runs a more benign course in children during the acute phase. Unlike adult patients, no fatalities were reported among the paediatric SARS patients. Published data on long-term sequelae of SARS are very limited. In our follow-up study, although patients have clinically recovered from their initial illness, exercise impairment and residual radiological abnormalities were demonstrated at 6 months after diagnosis. It is important to assess these patients on a regular basis to detect and provide appropriate management for persistent or emerging long-term sequelae in the physical, psychological and social domains. PMID- 15531254 TI - SARS: future research and vaccine. AB - Severe acute respiratory syndrome (SARS) is a new infectious disease of the 21st century that has pandemic potential. A novel coronavirus (CoV) was identified as its aetiological agent and its genome was sequenced within months of the World Health Organisation issuing a global threat on SARS. The high morbidity and mortality of this potentially pandemic infection demands a rapid research response to develop effective antiviral treatment and vaccine. This will depend on understanding the pathogenesis and immune response to SARS CoV. Further understanding of the ecology of SARS CoV in human and animals will help prevent future cross species transmission. Likewise for the super-spreading events, clarification of the underlying reasons will be important to prevent a large scale outbreak of SARS. Lastly it is of utmost importance that international research collaboration should be strengthened to deal with SARS and any other emerging infectious disease that can seriously threaten our future. PMID- 15531255 TI - Educating health professionals to improve quality of care for asthma. AB - Recent studies have shown significant progress in improving the quality of care for asthma. The most successful interventions have combined multiple elements: educational sessions that engage learners in discussing cases and practicing new skills; use of new resources, tools and practice patterns to enable quality improvement; and reinforcement of improvements through peer support, incentives or administrative review. Future research should include more randomised controlled trials to test the effectiveness of quality improvement interventions, more detailed descriptions of strategies to change health professional behaviour and studies to determine whether effective interventions can be translated to other settings, disseminated widely and sustained over time. PMID- 15531256 TI - Respiratory manifestations in HIV-infected children pre- and post-HAART in Abidjan, the Ivory Coast. AB - Among children infected with human immunodeficiency virus (HIV), respiratory diseases are a frequent cause of morbidity and mortality. This review describes respiratory manifestations of paediatric HIV infection before and after the beginning of HAART in Abidjan, Ivory Coast. In an observational cohort, HIV infected children had quarterly clinical visits and a day-clinic available all week for ill children. CD4 and viral load were measured at baseline and every 6 months thereafter. All children with a CD4 percentage below 25% were prescribed daily cotrimoxazole prophylaxis. Ninety-eight children (of a total of 282) were recruited before HAART and treated during the follow-up, there were 56 boys and 42 girls, with a mean age of 6.2 years at inclusion. The mean percentage of CD4 before HAART was 8.7%. Twelve children had a history of pulmonary tuberculosis and five were on antituberculosis treatment at inclusion. Fifty-one per cent presented with abnormalities on chest X-ray at inclusion. Before initiation of HAART, respiratory manifestations represented 32.4% of morbidity events and the incidence for 100 child/months was 9.29 for URTI, 15.2 for bronchitis, 6.07 for LRTI, 0.71 for tuberculosis and 0.36 for Pneumocystis carinii. After the initiation of HAART, respiratory manifestations represented 40.9% of all morbidity events and the incidence for 100 child/months was 5.35 for URTI, 9.48 for bronchitis, 2.17 for LRTI and 0.16 for tuberculosis. During HAART treatment, the incidence of respiratory infections decreased dramatically compared to before the antiretroviral treatment. However, respiratory events still represented 40% of all events occurring following the start of HAART therapy. PMID- 15531257 TI - Pulmonary alveolar proteinosis in children. AB - Pulmonary alveolar proteinosis (PAP) is a rare cause of chronic interstitial lung disease in children characterised by accumulation of a lipoproteinaceous material in the alveoli. Bronchoalveolar lavage is the key diagnostic tool, revealing a milky appearance of the return fluid and a periodic acid-Schiff staining material in the alveolar macrophages. PAP is a heterogeneous disease. Immediate-onset forms leading to early and fatal respiratory failure may be related to SP-B deficiency. Postnatal-onset PAP may be associated with various diseases or may be primary. The latter has a polymorphic progression from asymptomatic to uncontrollable respiratory failure. Recent studies have implicated GM-CSF and/or its receptor but the exact underlying mechanisms are still unknown. Therapeutic lung lavages are the only effective treatment for severe cases. PMID- 15531258 TI - High frequency oscillatory ventilation in acute respiratory failure. AB - High frequency oscillatory ventilation (HFOV) has emerged over the past 20 years as a safe and effective means of mechanical ventilatory support in patients with acute respiratory failure. During HFOV, lung recruitment is maintained by application of a relatively high mean airway pressure with superimposed pressure oscillations at a frequency of 3 to 15Hz, creating adequate ventilation using tidal volumes less than or equal to the patient's dead space volume. The physiologic rationale for the application of HFOV in the clinical arena comes from its ability to preserve end-expiratory lung volume while avoiding parenchymal overdistension at end-inspiration and theoretically limiting the potential for ventilator-associated lung injury. Data in the neonatal population suggests significant benefits in pulmonary outcomes when HFOV is applied with a recruitment strategy in preterm infants with respiratory distress syndrome (RDS). Use of HFOV in the paediatric and adult populations has not as yet been associated with significant improvements in clinically important outcome measures. PMID- 15531259 TI - Do environmental pollutants influence the onset of respiratory syncytial virus epidemics or disease severity? AB - The trigger for annual epidemics of respiratory syncytial virus (RSV) infections and the factors contributing to the disappearance of RSV infections in late winter remain obscure. Similarly, there is no adequate explanation for the higher morbidity and admission rates in industrialised as compared with rural areas. It has been suggested that a variety of environmental factors such as temperature, daylight and humidity may influence the onset and waning of the epidemics. However, the few studies assessing these variables fail to support such a link. In many tropical countries the annual epidemic occurs in the summer or autumn, arguing against temperature having a direct influence. A number of studies have suggested that indoor pollutants, including cigarette smoke, are associated with an increased likelihood of being admitted to hospital with severe lower respiratory tract disease. One study exploring the potential role of outdoor pollutants on the pattern of RSV related illness in infancy was unable to identify a clear link between a variety of pollutants and the timing of the epidemic. Nitric oxide levels were higher in winter than during the summer and much higher winter peaks of NO were observe in industrialised areas as compared with urban and rural areas. Whether this or other environmental pollutants contribute to the higher incidence of severe disease in industrialised areas is unclear. Further work is required to explore the possible influence of NO and other environmental pollutants on both the timing and severity of epidemics. PMID- 15531260 TI - Thirty Years of Statins, Festschrift in honor of Dr. Akira Endo. PMID- 15531272 TI - Statins have offered great benefits. 2002. PMID- 15531273 TI - Competitive inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase by ML 236A and ML-236B fungal metabolites, having hypocholesterolemic activity. 1976. PMID- 15531274 TI - Therapeutic effects of ml-236B in primary hypercholesterolemia. 1980. PMID- 15531275 TI - Effects of an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme a reductase on serum lipoproteins and ubiquinone-10 levels in patients with familial hypercholesterolemia. 1981. PMID- 15531276 TI - Lowering plasma cholesterol by raising ldl receptors. 1981. PMID- 15531277 TI - Mevinolin and colestipol stimulate receptor-mediated clearance of low density lipoprotein from plasma in familial hypercholeserolemia heterozygotes. 1983. PMID- 15531278 TI - The discovery and development of HMG-CoA reductase inhibitors. 1992. PMID- 15531279 TI - Randomised trial of cholesterol lowering in 4444 patients with coronary heart disease: the Scandinavian Simvastatin Survival Study (4S). 1994. PMID- 15531280 TI - Lower patient's cholesterol now. Trial evidence shows clear benefits from secondary prevention. 1995. PMID- 15531281 TI - Prevention of coronary heart disease with pravastatin in men with hypercholesterolemia. 1995. PMID- 15531283 TI - Goals of statin therapy: three viewpoints. 2002. PMID- 15531282 TI - Lipoprotein changes and reduction in the incidence of major coronary heart disease events in the Scandinavian Simvastatin Survival Study (4S). 1998. PMID- 15531284 TI - Guidelines for lowering lipids to reduce coronary artery disease risk: a comparison of rosuvastatin with atorvastatin, pravastatin, and simvastatin for achieving lipid-lowering goals. 2003. PMID- 15531285 TI - The origin of the statins. 2004. PMID- 15531289 TI - Evaluation and comparison of commercially available Aloe vera L. products using size exclusion chromatography with refractive index and multi-angle laser light scattering detection. AB - Raw materials supplied as Aloe vera L. (sometimes referred to as Aloe barbadensis) samples often contain different composition of low and high molecular weight components when analyzed by size exclusion chromatography. One major reason for variable compositions of commercial A. vera L. materials is that they are produced by different manufacturing techniques. Consistent composition of matter based upon a given standard has been difficult to define. In addition, the method of quantifying and characterization of these commercially available materials has not been agreed upon within the industry. The end user, whether a researcher, a manufacturer, a marketing arm of industry or the consumer, should know that they are receiving a consistent product. A blind study of 32 various A. vera L. samples from different manufacturers, and a prepared sample of fresh A. vera L. gel with the commercial, biologic drug Acemannan Immunostimulanttrade mark, were analyzed for content of high molecular weight (polysaccharides) material by size exclusion chromatography with refractive index detection (SEC/RI) and SEC/RI coupled with multi-angle laser light scattering (MALLS) detection. Results from the SEC/RI analysis showed significant variation in the high molecular weight content, and the MALLS analysis also showed significant variation versus SEC/RI. In addition, HPLC analysis of the anthraquinone content showed that all samples contained significantly less than that of the raw, unwashed aloe gel. The variation of results from all analysis is attributed to differing methods in which the samples were processed by the different manufacturers. PMID- 15531290 TI - Analysis of the anthraquinones aloe-emodin and aloin by gas chromatography/mass spectrometry. AB - A procedure was developed for the determination of low levels of the anthraquinones aloe-emodin and aloin A (barbalin) in aloe products based on gas chromatography/mass spectrometry (GC/MS) of the trimethyl silyl (TMS) derivatives of these analytes in the presence of Chrysophanol used as internal standard. The method was used to analyze several aloe based commercial products (liquids, gels and solids). Wide variation in the level of these anthraquinones was observed among the different products. The method had a sensitivity of 0.005 ppm of aloe emodin and 0.05 ppm of aloin. PMID- 15531291 TI - Isolation and characterization of structural components of Aloe vera L. leaf pulp. AB - The clear pulp, also known as inner gel, of Aloe vera L. leaf is widely used in various medical, cosmetic and nutraceutical applications. Many beneficial effects of this plant have been attributed to the polysaccharides present in the pulp. However, discrepancies exist regarding the composition of pulp polysaccharide species and an understanding of pulp structure in relation to its chemical composition has been lacking. Thus, we examined pulp structure, isolated structural components and determined their carbohydrate compositions along with analyzing a partially purified pulp-based product (Acemannan hydrogel) used to make Carrisyn hydrogel wound dressing. Light and electron microscopy showed that the pulp consisted of large clear mesophyll cells with a diameter as large as 1000 microm. These cells were composed of cell walls and cell membranes along with a very limited number of degenerated cellular organelles. No intact cellular organelles were found in mesophyll cells. Following disruption of pulp by homogenization, three components were isolated by sequential centrifugation. They were thin clear sheets, microparticles and a viscous liquid gel, which corresponded to cell wall, degenerated cellular organelles and liquid content of mesophyll cells based on morphological and chemical analysis. These three components accounted for 16.2% (+/-3.8), 0.70% (+/-0) and 83.1% of the pulp on a dry weight basis. The carbohydrate composition of each component was distinct; liquid gel contained mannan, microparticles contained galactose-rich polysaccharide(s) and cell walls contained an unusually high level of galacturonic acid (34%, w/w; Gal A). The same three components were also found in Acemannan Hydrogel with mannan as the predominant component. Thus, different pulp structural components are associated with different polysaccharides and thus may potentially be different functionally. These findings may help lay a basis for further studies and development of better controlled processing methods and applications for this well-accepted medicinal plant. PMID- 15531292 TI - Fractionation of Aloe vera L. inner gel, purification and molecular profiling of activity. AB - Products derived from the inner gel of the Aloe vera L. plant have demonstrated multiple clinical activities, and are used routinely to accelerate wound healing. However, typical of natural products, the complex nature of Aloe vera gels may contribute to diverse pharmacologic activities. Our focus on the hematopoietic activities of Aloe vera extracts is extended by these functional studies, which used purified fractions from Aloe vera gel and included a preliminary organ specific in vitro molecular profile. Studies using a >99% pure carbohydrate fraction from Aloe vera extracts revealed increased hematopoietic and hematologic activity compared to the starting material. In addition, this fraction differentially regulated liver and lung cytokine mRNA levels, resulting in significant increases in message for hematopoietic cytokines [granulocyte colony stimulating factor (G-CSF) and stem cell factor (SCF)]. This profile of activity differed from another fraction obtained from Aloe vera, suggesting the potential for diverse pharmacologic activity. The molecular studies were undertaken using co-cultures of organ slices to limit the amount of purified material required. In summary, these studies revealed significant hematopoietic activity by both pharmacologic and molecular analysis using a >99% pure carbohydrate fraction from Aloe vera gels. PMID- 15531293 TI - Aloe-emodin modulates PKC isozymes, inhibits proliferation, and induces apoptosis in U-373MG glioma cells. AB - Aloe-emodin (1,8-dihydroy-3-[hydroxymethyl]-anthraquione) purified from Aloe vera leaves has been reported to have antitumor activity. The objectives of our research were to determine how aloe-emodin regulates the cell cycle, cell proliferation and protein kinase C (PKC) during glioma growth and development. To establish the cell cycle effects of aloe-emodin on brain cells [transformed glia cell line (SVG) and human glioma U-373MG cell line (U-373MG)], cells were treated with either dimethylsulfoxide (DMSO; control) or aloe-emodin (40 microM). Results from flow cytometry demonstrated that aloe-emodin delayed the number of cells entering and exiting DNA synthesis (S) phase in both SVG and U-373MG cells indicating that aloe-emodin may inhibit S phase progression. Assessment of cell viability demonstrated that SVG and U-373MG glioma cell were highly sensitive to aloe-emodin. The aloe-emodin-induced decreased proliferation was sustained at 48 96 h. A PKC activity assay was quantified to establish the role of PKC in aloe emodin's mode of action. Exposure of SVG and U-373MG glioma cells to aloe-emodin suppressed PKC activity and reduced the protein content of most of the PKC isozymes. We determined that cancer growth inhibition by aloe-emodin was due to apoptosis (i.e., programmed cell death). Taken together, these results support the hypothesis that aloe-emodin represents a novel antitumor chemotherapeutic drug. PMID- 15531294 TI - Visualization of naturally occurring Foxp3+ regulatory T cells in normal and tumor-bearing mice. AB - CD25+CD4+ regulatory T cells (Treg) play pivotal roles in the host response to tumors. However, their exact location in vivo is largely unknown. The forkhead/winged helix transcription factor, Foxp3, is specifically expressed in naturally occurring Treg (nTreg) and programs their development and function. In this study, we produced a rabbit polyclonal antibody (pAb), which can detect mouse Foxp3 protein in situ. Results using this pAb revealed that Foxp3+CD4+ nTreg cells occur in direct contact with CD11c+ dendritic cells (DCs), and Foxp3 CD4+ and CD8+T lymphocytes in the T cell regions of lymphoid tissues from normal and tumor-bearing mice. The numbers of Foxp3+CD4+ nTreg cells are significantly increased in draining, but not nondraining, lymph nodes (LNs) and spleen (SPL) of tumor-bearing mice. Furthermore, a small number of nTreg could be also found at the tumor site. These observations support the notion that the numbers of Foxp3+CD4+ nTreg are increased by tumors and may contribute to the immunosuppression observed in tumor-bearing hosts at secondary lymphoid organs and also possibly at the tumor site. PMID- 15531295 TI - Piperine is a potent inhibitor of nuclear factor-kappaB (NF-kappaB), c-Fos, CREB, ATF-2 and proinflammatory cytokine gene expression in B16F-10 melanoma cells. AB - Immune regulation, induction of various inflammatory and growth regulatory genes such as IL-1beta, IL-6, TNF-alpha and GM-CSF require activation of transcription factors such as nuclear factor-kappaB (NF-kappaB), activated transcription factor (ATF-2), c-Fos and cAMP response element-binding protein (CREB). Untreated B16F 10 cells produce very high amount of proinflammatory cytokines such as IL-1beta, IL-6, TNF-alpha and GM-CSF. Piperine treatment significantly reduced the above proinflammatory cytokines. We also found that piperine could reduce the expression of IL-1beta, IL-6, TNF-alpha, GM-CSF and IL-12p40 genes. Piperine at a concentration of 2.5, 5 and 10 microg/ml inhibited the collagen matrix invasion of B16F-10 melanoma cells in a dose-dependent manner. Piperine could inhibit the matrix metalloproteinase production which was demonstrated by zymographic analysis. We found that the nuclear translocation of p65, p50, c-Rel subunits of NF-kappaB and other transcription factors such as ATF-2, c-Fos and CREB were inhibited by the treatment of piperine. PMID- 15531296 TI - Morphine aggravates the apoptosis of simian immunodeficiency virus infected CEM x174 cells in the prolonged culture in vitro. AB - This study was designed to assess the in vitro effects of morphine on the lymphocytes infected with SIV. CEM x174 cells were cotreated with morphine and simian immunodeficiency virus (SIVmac239). Cells were cultured for 96 h and the effects of morphine on the viability of infected cells were determined. At the concentration of 1 micromol/l, morphine could inhibit the proliferation of CEM x174 cells at the culture of 72 h. The stronger effect was observed in the case of viral infection. During 72 h SIV loading, the cells were accumulated in S phase in all SIV infected groups. The S arrest was observed in every experimental group and statistically different from normal groups (P<0.05). The results from annexin V binding assay showed that SIV infection resulted in a lower proportion of vital cells and higher mortality compared with corresponding control (P<0.01). Morphine failed to induce detectable alteration in the cell cycle profile of viral infected cells. Western blotting showed that the synthesis of intracellular p53 and bax protein was gradually up-regulated in the virus-loading period of 72 h. Naloxone had an apparent additive rather than antagonistic effect on the morphine-associated enhancement of bax expression. The ratio of bax/bcl-2 proteins appeared to tilt the balance toward apoptosis. At 72 h of infection, 1 micromol/l of morphine significantly elevated the level of caspase-3. These results indicated that the alteration in the balance of intracellular apoptotic and anti-apoptotic elements is one of the reasons of accelerated progression of acquired immunodeficiency syndrome (AIDS) by opioids abuse. PMID- 15531297 TI - Administration of antisense phosphorothioate oligonucleotide to the p65 subunit of NF-kappaB inhibits established asthmatic reaction in mice. AB - The transcription factor, nuclear factor (NF)-kappaB, which transactivates various genes for proinflammatory cytokines and many other immunoregulatory genes, plays an important role in the regulation of various inflammatory diseases including asthma. Its increased activation has been demonstrated in the lungs after allergen challenge and in airway epithelial cells and macrophages of asthmatic patients. In the present study, we investigated whether the pretreatment with p65 antisense results in a significant inhibition of asthmatic reactions in a mouse model. Mice sensitized and challenged with ovalbumin (OVA) showed typical asthmatic reactions as follows: (1) an increase in the number of eosinophil in bronchoalveolar lavage fluid; (2) a marked influx of inflammatory cells into the lung around blood vessels and airways, and airway luminal narrowing; (3) the development of airway hyperresponsiveness; (4) the detection of TNF-alpha and Th2 cytokines, such as IL-4 and IL-5 in the bronchoalveolar lavage (BAL) fluid; and (5) detection of allergen-specific IgE and IgG in the serum. Two successive intravenous administration of p65 antisense before the last airway OVA challenge resulted in a significant inhibition of all asthmatic reactions, whereas the p65 nonsense did not produce such effects. In addition, the p65 antisense inhibition of asthmatic reaction appears to be due to the initial suppression of an allergen-specific IgE response, inducing degranulation of mast cells through the cross-linking of allergen-specific IgE. This study may provide evidence that NF-kappaB plays a critical role in the pathogenesis of asthma in mice. PMID- 15531298 TI - Identification of a host gene subset related to disease prognosis of HIV-1 infected individuals. AB - Although plasma-virus-RNA level and CD4-positive-T-cell count are useful to monitor clinical status of the human immunodeficiency virus (HIV)-infected individuals, clinical course is often varied among patients and sometimes difficult to predict. To identify additional parameters associated with disease progression, we examined by cDNA microarray the expression profiles of 731 immune response-related genes in the peripheral blood mononuclear cells (PBMCs) from 21 HIV-positive individuals in Uganda. The analysis enabled the patients to be classified into three distinct groups on the basis of the gene expression patterns. Notably, these groups, clusters I, II and III, were highly associated with clinical status of the patients defined by CDC classification, categories A, B, and C, respectively. Statistical analysis identified 40 genes whose expressions were significantly up- or down-regulated in the cluster III patients (p<0.05). Up- and down-regulated genes included ones involved in immature T lymphocytes differentiation, apoptosis signaling, and active HIV replication, suggesting that the levels of active destruction and regeneration of mature T lymphocytes associated with enhanced HIV-1 replication is related to the disease progression. Follow-up study showed that the cluster classification improved prediction of disease prognosis with the CDC classification. These findings provide new clues for studying perturbation of host immunity, pathogenesis, and disease prognosis of HIV-infected individuals. PMID- 15531299 TI - Lack of nitric oxide-mediated regulation of amylase secretion stimulated by VIP in parotid glands of NOD mice. AB - The non-obese diabetic (NOD) mouse is chosen among other experimental models to study autoimmune sialadenitis resembling Sjogren's syndrome (SS), because of its unique characteristic of developing salivary dysfunction. Based on the deep loss of nitric oxide synthase (NOS) activity in parotid glands of NOD mice observed from early stages of disease and the inhibitory effect of nitric oxide (NO) donors on amylase secretion in normal salivary glands, our goal was to investigate whether parotid glands from NOD mice lacking NOS activity presented this regulatory mechanism of amylase secretion. We found that parotid glands from NOD mice lack nitric oxide-mediated regulation of amylase secretion in response to VIP stimulation. The lack of regulation might be assigned to the loss of NOS activity as derived from the results with NOS inhibitors and increasing concentrations of VIP. These functional differences observed in NOD vs. BALB/c parotid glands occur in the absence of immune infiltrates in exocrine tissue, and it is not related to cAMP accumulation. NO-mediated regulation of amylase secretion was not observed in BALB/c submandibular glands to the same extent as described in parotid glands and was absent in submandibular glands of NOD mice. PMID- 15531300 TI - Characterization of (2R, 3S)-2-([[4-(2-butynyloxy)phenyl]sulfonyl]amino)-N,3 dihydroxybutanamide, a potent and selective inhibitor of TNF-alpha converting enzyme. AB - TNF-alpha converting enzyme (TACE) is a validated therapeutic target for the development of oral tumor necrosis factor-alpha (TNF-alpha) inhibitors. Here we report the pre-clinical results and characterization of a selective and potent TACE inhibitor, (2R, 3S)-2-([[4-(2-butynyloxy)phenyl]sulfonyl]amino)-N,3 dihydroxybutanamide (TMI-2), in various in vitro and in vivo assays. TMI-2 is a potent TACE inhibitor in an enzymatic FRET assay (IC50=2 nM). It is more than 250 fold selective over MMP-1, -7, -9, -14, and ADAM-10 in vitro. In cell-based assays and human whole blood, TMI-2 inhibits lipopolysaccharide (LPS)-induced TNF secretion with IC50s<1 uM. Importantly, TMI-2 inhibits the spontaneous release of TNF-alpha in human synovium tissue explants of rheumatoid arthritis patients with an IC50 of 0.8 microM. In vivo, TMI-2 potently inhibits LPS-induced TNF-alpha production in mice (ED50=3 mg/kg). In the adjuvant-induced arthritis (AIA) model in rats, treatment with TMI-2 at 30 mg/kg and 100 mg/kg p.o. b.i.d. was highly effective in reducing joint arthritis scores. In a semi-therapeutic collagen induced arthritis (CIA) model in mice, TMI-2 is highly effective in reducing disease severity scores after oral treatment at 100 mg/kg twice per day. In summary, TMI-2 is a potent and selective TACE inhibitor that inhibits TNF-alpha production and reduces the arthritis scores in pre-clinical models. TMI-2 represents a novel class of TACE inhibitors that may be effective and beneficial in the treatment of rheumatoid arthritis as well as other TNF-mediated inflammatory autoimmune diseases. PMID- 15531301 TI - Characterization of an interaction between fetal hemoglobin and lipid A of LPS resulting in augmented induction of cytokine production in vivo and in vitro. AB - A previously described extract of sheep fetal liver was reported to reverse many of the cytokine changes associated with aging in mice, including an augmented spleen cell ConA-stimulated production of IL-4 and decreased production of IL-2. Similar effects were not seen with adult liver preparations. These changes were observed in various strains of mice, including BALB/c, DBA/2 and C57BL/6, using mice with ages ranging from 8 to 110 weeks. Preliminary characterization of this crude extract showed evidence for the presence of Hb gamma chain, as well as of lipid A of LPS. We show below that purified preparations of sheep fetal Hb, but not adult Hb, in concert with suboptimally stimulating doses of LPS (lipid A), cooperate in the regulation of production of a number of cytokines, including TNFalpha and IL-6, in vitro. Furthermore, isolated fresh spleen or peritoneal cells from animals treated in vivo with the same combination of Hb and LPS, showed an augmented capacity to produce these cytokines on further culture in vitro. Evidence was also obtained for a further interaction between CLP, LPS and fetal Hb itself in this augmented cytokine production. These data suggest that some of the functional activities in the fetal liver extract reported earlier can be explained in terms of a novel immunomodulatory role of a mixture of LPS (lipid A) and fetal Hb. PMID- 15531302 TI - Possible involvement of mast cells in collagen remodeling in the late phase of cutaneous wound healing in mice. AB - We examined possible roles of mast cells in cutaneous wound healing using mast cell deficient (W/Wv) mice and their normal littermates (+/+). A round full thickness wound was made on the back skin of these mice. The wounds closed completely within 20 days, and there was no difference in wound contraction between +/+ and W/Wv mice during the wound healing. While either chymase or tryptase activities were hardly detectable in W/Wv mice, chymase activities decreased at the impaired sites and recovered to the control level within 20 days in +/+ mice. Tryptase activities were higher than the control level on day 15 and day 20 in +/+ mice. Histological observations on day 15 and day 20 in +/+ mice revealed that mast cells were abundant at the wound edges but absent at the center. The latent and the active forms of MMP-2 and MMP-9 increased on day 10 and day 15 but recovered nearly to control levels on day 20 in both mice groups. The hydroxyproline contents in W/Wv mice were significantly higher than those in +/+ mice on day 15 and day 20. Furthermore, histological observations revealed that the collagen aggregation at the wound edges was tighter and less interwoven in W/Wv mice compared with +/+ mice. These results suggest that mast cells accumulated at the wound edge may participate in tissue remodeling in the late phase of wound healing. PMID- 15531303 TI - Prediction of postoperative visual outcome based on hole configuration by optical coherence tomography in eyes with idiopathic macular holes. AB - PURPOSE: To evaluate whether an index based on hole configuration can be used to predict visual outcome in eyes with idiopathic macular holes. DESIGN: Prospective interventional case series. METHODS: Thirty-five eyes of 32 patients with idiopathic stage 2 or 3 macular hole were enrolled in this study. The best corrected visual acuity (BCVA), cross-sectional image of the macular hole by optical coherence tomography (OCT), and retinal thickness in the central (<1000 microm), inner (1000 to 2220 microm), and outer ring areas (2220 to 3450 microm) as defined by the OCT retinal mapping program were evaluated preoperatively and at 1, 3, 6, and 12 months postoperatively. The change in retinal thickness of the inner ring area at the 6-month postoperative period was used to evaluate the degree of preoperative retinal deformation. The macular hole index (MHI) (ratio of hole height to base diameter of hole) was calculated and correlated with minimum diameter of hole, base diameter of hole, the postoperative decrease in macular thickness, and the postoperative BCVA. The postoperative BCVA was further evaluated in two patient-matched groups. RESULTS: Retinal thickness values in the inner ring area were decreased at the 1-month postoperative period. MHI significantly correlated with the postoperative decrease in macular thickness in the inner ring area at 6 months (correlation coefficient = -0.632, P = .030, Spearman analysis) and with the postoperative BCVA (P = .013, multiple regression analysis). Postoperative BCVA in the MHI >/=0.5 group was better than that in the MHI <0.5 group (P = .032, Mann-Whitney test). CONCLUSIONS: The MHI is a ratio easily calculated from OCT transverse images of the macular area. The MHI represents the preoperative configuration of a macular hole and is a prognostic factor for visual outcome. PMID- 15531304 TI - Central corneal thickness and measured IOP response to topical ocular hypotensive medication in the Ocular Hypertension Treatment Study. AB - PURPOSE: To determine whether central corneal thickness (CCT) correlates with measured intraocular pressure (IOP) response to topical ocular hypotensive medication in the Ocular Hypertension Treatment Study (OHTS). DESIGN: Prospective randomized clinical trial. METHODS: Intraocular pressure measurements were performed by Goldmann applanation tonometry. Central corneal thickness was measured by ultrasonic pachymetry. The following indicators of IOP response to topical ocular hypotensive medication were examined: (1) IOP after an initial four- to six-week one-eyed therapeutic trial of a nonselective beta-blocker (N = 549) or a prostaglandin analog (N = 201); (2) the mean IOP response during 12 to 60 months of follow-up among medication participants (N = 689); (3) the percentage of follow-up visits at which both eyes met the treatment goal; (4). the total number of different medications prescribed to reach treatment goal; and (5) the total number of different medications prescribed multiplied by the number of months each medication was prescribed. RESULTS: Central corneal thickness was inversely related to the IOP response after the initial one-eyed therapeutic trial and during 12 to 60 months of follow-up (P < .05). Mean CCT was not correlated with the number of different medications prescribed during follow-up, the total medication-months, or the percentage of visits at which IOP target was met. CONCLUSIONS: Individuals with thicker corneas had smaller measured IOP responses to ocular hypotensive medication than those with normal or thin corneas. We believe that CCT measurements may be useful in patient management and in interpreting clinical trials of ocular hypotensive medication. PMID- 15531305 TI - Position of angle-supported, iris-fixated, and ciliary sulcus-implanted myopic phakic intraocular lenses evaluated by Scheimpflug photography. AB - PURPOSE: To examine postoperative positional stability of myopic phakic intraocular lenses (IOLs). DESIGN: Prospective, nonrandomized clinical study. METHODS: The study included 46 eyes which received an anterior chamber angle supported (Bausch & Lomb NuVita; 10 eyes), anterior chamber iris-fixated (Ophtec Artisan; 20 eyes) or ciliary sulcus-implanted phakic IOL (Staar ICL; 16 eyes). The distance between the phakic IOL and the crystalline lens and the cornea as well as rotation around the optical axis was evaluated using Scheimpflug photography at 1, 3 to 6, and 12 months postsurgery. RESULTS: The anterior chamber phakic IOLs showed no significant movement in anteroposterior direction. The posterior chamber phakic IOL showed a significant movement toward the crystalline lens between postoperative months 3 and 12. The median amount of rotation around the optical axis between the 3- and the 12-month evaluation was 1.9 degrees (range = 0.0-33.5 degrees) for the NuVita, 0.6 degrees (range = 0.0 3.5 degrees) for the Artisan, and 0.9 degrees (range = 0.2-2.3 degrees) for the ICL. Four NuVita IOLs rotated more than 10 degrees. CONCLUSIONS: The angle supported anterior chamber phakic IOLs showed a generally stable position regarding distance to cornea and natural lens, but rotation was observed in four IOLs. The iris-fixated phakic IOL showed the highest overall stability. The posterior chamber phakic IOL was stable in terms of rotation but had a tendency to decrease in distance toward the crystalline lens. Intraocular lenses implanted in phakic eyes followed for 12 months demonstrate stable IOL position overall. PMID- 15531306 TI - Diagnosis of macular pseudoholes and lamellar macular holes by optical coherence tomography. AB - PURPOSE: To assess the usefulness of optical coherence tomography (OCT) for better differential diagnosis of macular pseudoholes (MPH) and lamellar macular holes (LMH). DESIGN: Observational case series. METHODS: setting: Institutional practice. patients: We reviewed the files of 71 eyes of 70 consecutive patients who were diagnosed as having a pseudohole or lamellar hole on OCT examination. All patients referred for suspected pseudohole or lamellar hole on biomicroscopy were evaluated by OCT. main outcome measures: Each eye underwent six radial 3-mm OCT scans centered on the macula, one 6-mm vertical and one 6-mm horizontal scan. Retinal thickness was measured at the foveal center and 750 microm from the center, vertically, and horizontally. The diameter of the macular contour was also measured on vertical and horizontal scans. RESULTS: In 40 cases, OCT showed a macular profile characteristic of MPH: a steepened foveal pit combined with thickened foveal edges and a small foveal pit diameter. Central foveal thickness was normal or slightly increased (167 +/- 42 microm). Mean perifoveal thickness was greater than normal (363 +/- 65 microm). In 29 other cases corresponding to LMH, OCT showed a profile characterized by a thin irregular foveal floor, split foveal edges, and near-normal perifoveal retinal thickness. Central foveolar thickness was thinner than normal (72 +/- 19 microm). Mean perifoveal thickness was near normal (283 +/- 36). Optical coherence tomography did not allow the classification of the remaining two cases. CONCLUSIONS: Optical coherence tomography is very useful in distinguishing MPH attributable to epiretinal membrane contraction from LMH because of partial opening of a macular cyst. PMID- 15531307 TI - Intravitreal triamcinolone acetonide and intraocular pressure. AB - PURPOSE: To analyze the incidence of intraocular pressure (IOP) elevation following intravitreal triamcinolone injection. DESIGN: Retrospective observational case series. METHODS: Charts of patients undergoing intravitreal triamcinolone injection in one clinical practice were reviewed. A pressure elevation was defined as a pressure of 24 mm Hg or higher during follow-up. RESULTS: There were 89 patients with a mean age of 76.4 years. The mean baseline IOP was 14.9 mm Hg with a mean change of 8.0 mm Hg. Thirty-six patients (40.4%) experienced a pressure elevation to 24 mm Hg or higher at a mean of 100.6 days (SD = 83.1 day) after treatment. Of nonglaucomatous patients with baseline IOP of 15 mm Hg or above, 60.0% experienced a pressure elevation, compared with only 22.7% of those with baseline pressures below 15 mm Hg (relative risk = 2.1, P < .01). In glaucoma patients, 6 of 12 (50%) experienced a pressure elevation, and this elevation was not correlated with baseline pressure. Thirty-two patients (36.0%) received repeat injections, and there was no difference in the incidence of procedure elevation in patients receiving multiple injections versus those receiving a single injection. Pressure elevation was controlled with topical medications in all patients. CONCLUSIONS: IOP elevation after intravitreal triamcinolone injection is common and may take an extended period of time to manifest. The proportion of patients who developed a pressure elevation to at least 24 mm Hg was much higher for those with baseline IOP 15 mm Hg or greater. PMID- 15531308 TI - Corneal thickness in children. AB - OBJECTIVE: To determine normal central and paracentral corneal thickness measurements in the pediatric population and to determine if these measurements are consistent across different pediatric age groups and different racial groups. DESIGN: Prospective observational case series. METHODS: Pachymetry measurements were performed on 198 eyes of 108 children. The measurements were taken centrally as well as at four paracentral sites 3 mm from the corneal center at the 3, 6, 9, and 12 o'clock positions. The two-tailed t test was used for comparison of the continuous means for values of corneal thickness. Analysis of variance (ANOVA) was performed to determine differences among age and ethnic groups RESULTS: The mean central corneal thickness (CCT) was 549 +/- 46 microm. Paracentral corneal thickness mean values, as measured 3 mm from the corneal center, were as follows: superior, 575 +/- 52 microm; nasal, 568 +/- 50 microm; inferior, 568 +/- 51 microm; and temporal, 574 +/- 47 microm. The mean CCT values were significantly thinner than at each of the mean paracentral points (P < .05 for each comparison, paired t test). Paracentral corneal thickness measurements demonstrated no significant differences between locations (P > .05, variance analysis). The mean CCT +/- SD for each age group was as follows: 6 to 23 months, 538 +/- 40 microm; 2 to 4 years, 546 +/- 41 microm; 5 to 9 years, 566 +/- 48 microm; and 10 to 18 years, 554 +/- 35 microm (ANOVA P = .012). ANOVA performed on central pachymetry values demonstrated no significant differences among racial subgroups. CONCLUSIONS: Pediatric central and paracentral corneal thicknesses increase slowly over time and reach adult thicknesses at 5 to 9 years of age. PMID- 15531309 TI - Dominant optic atrophy, sensorineural hearing loss, ptosis, and ophthalmoplegia: a syndrome caused by a missense mutation in OPA1. AB - PURPOSE: To describe the clinical features of and identify the disease-causing mutation in a large Utah family segregating a dominantly inherited syndrome of optic atrophy, sensorineural hearing loss, ptosis, and ophthalmoplegia. DESIGN: Observational case series. METHODS: Thirty individuals at risk for a syndrome of optic atrophy, sensorineural hearing loss, ptosis, and ophthalmoplegia in a single family underwent clinical examinations and venipuncture. Linkage analysis and mutation screening of the optic atrophy 1 gene (OPA1) were performed. RESULTS: Eighteen individuals demonstrated characteristics of the syndrome. Genetic analysis identified a G-->A substitution at nucleotide position 1334 in exon 14 of OPA1 causing an arginine-to-histidine change (R445H) in all affected members of the family. This change segregated with the disease phenotype in the study family with a LOD score of 7.02 at theta; = 0 and was not found in 200 normal control subjects. Analysis of an unrelated Belgian family with a similar phenotype revealed the same R445H mutation segregating with the disease phenotype. CONCLUSIONS: This study describes a mutation in OPA1 causing a unique syndrome of optic atrophy, sensorineural hearing loss, ptosis, and ophthalmoplegia. These results expand the spectrum of human disease associated with mutations of OPA1 and indicate that ophthalmologists caring for patients with optic atrophy should inquire about possible associated hearing loss. Although OPA1 is a nuclear gene, the gene product localizes to mitochondria, suggesting that mitochondrial dysfunction may be the final common pathway for many forms of syndromic and nonsyndromic optic atrophy, hearing loss, and external ophthalmoplegia. PMID- 15531310 TI - Scanning electron microscopic studies of Supramid Extra from the patients displaying recurrent ptosis after frontalis suspension. AB - PURPOSE: To investigate the morphologic changes in Supramid Extra suture harvested from patients displaying recurrent ptosis who had undergone a frontalis suspension for congenital ptosis. DESIGN: Observational case series. METHODS: Using scanning electron microscopy, we examined the morphologic changes in Supramid Extra (S. Jackson Inc, Alexandria, Virginia, USA) harvested from 20 consecutive patients (23 eyelids) with recurrent ptosis who had undergone frontalis suspension. Disintegration was graded as mild, moderate, or severe. RESULTS: Examination of unused Supramid showed the smooth surface of the sheath and round, well-demarcated polyfilaments on cross section. However, implanted Supramid showed different degrees of disintegration. The surface of the sheath had a rough texture with cracks and peeling. On cross section, an accumulation of debris among the polyfilaments and irregular architectures with decreased diameters of the polyfilaments were noted in some cases. On the surface of polyfilaments, different degrees of debris were noted in all cases. The degree of the changes was not exactly correlated with either the time of the recurrence or the duration of implantation. However, as the duration of implanted Supramid increased, greater changes on the surface of the sheath were noted. With implantation of less than 37 months, mild changes (44%) were predominant; between 37 and 48 months, moderate changes (60%) were noted, and with more than 48 months of implantation, severe changes (44%) were predominant. CONCLUSIONS: Supramid undergoes morphologic degradation by hydrolysis after implantation. This change might be considered one of the possible factors that facilitate the recurrence of ptosis. PMID- 15531311 TI - Presurgical and postsurgical ultrasound assessment of lacrimal drainage dysfunction. AB - PURPOSE: In addition to the obligatory clinical tests, imaging of the lacrimal drainage system (LDS) is useful in its clinical evaluation. The purpose of this study was to examine the usability and reliability of ultrasonography in the evaluation of the lacrimal drainage system. DESIGN: Observational cohort study. METHODS: A prospective study was conducted at a single institution. We performed ultrasound examinations on 17 patients with epiphora before and after surgery, and on 17 asymptomatic volunteers, to visualize and evaluate the anatomic and functional condition or pathologic abnormalities of the LDS. RESULTS: Echographic evaluation of the LDS was possible in all individuals. Pathologic abnormalities (canaliculitis, diverticulitis, concretion, or dilation of the lacrimal sac, and reduced functionality of the orbicular muscle and/or lacrimal sac pump) could be well demonstrated. In the postsurgical course, functional patency of the dacryocystorhinostomy opening could be verified in all cases. CONCLUSIONS: Sonography of the LDS appears to represent a reliable diagnostic technique supplementary to clinical tests in the presurgical and postsurgical examination of patients with epiphora. Pathologic abnormalities that may not be apparent in routine x-ray dacryocystography can be demonstrated with ultrasound techniques. Patients also benefit from the avoidance of exposure to ionizing radiation. However, ultrasound is not suitable for imaging the lower part of the lacrimal sac and the lacrimal duct because of the presence of overlying bony structures. PMID- 15531312 TI - Lattice corneal dystrophy associated with the Ala546Asp and Pro551Gln missense changes in the TGFBI gene. AB - PURPOSE: To report a phenotypic variant of lattice corneal dystrophy associated with two missense changes, Ala546Asp and Pro551Gln, in the transforming growth factor-beta-induced gene (TGFBI). DESIGN: Experimental study. METHODS: Genomic DNA was obtained from the proband as well as affected and unaffected family members. Exons 4, 11, 12, and 14 of the TGFBI gene were amplified and sequenced. Additionally, a corneal button excised from the proband was examined by light and transmission electron microscopy. Haplotype analysis was performed on the proband's family and members of a previously identified pedigree with the same TGFBI gene missense changes. RESULTS: Bilateral, symmetric, radially arranged, branching refractile lines within and surrounding an area of central anterior stromal haze were noted in the proband. Multiple polymorphic, refractile deposits were noted in the mid and posterior stroma in both the proband and her daughter. Light and electron microscopic analyses demonstrated amyloid and excluded the presence of deposits characteristic of granular corneal dystrophy. Screening of TGFBI exon 12 in the proband and her affected daughter revealed two missense changes, Ala546Asp and Pro551Gln (both absent in 250 control chromosomes). Haplotype analysis suggested that the mutations in this family and in a previously identified pedigree reflect a founder effect, rather than an independent occurrence. CONCLUSIONS: We present a phenotypic variant of lattice corneal dystrophy associated with the Ala546Asp and Pro551Gln missense changes in exon 12 of the TGFBI gene. A common ancestor appears to account for the missense mutations observed in this pedigree and in a previously reported family. PMID- 15531313 TI - Visual experience during phacoemulsification under topical versus retrobulbar anesthesia: results of a prospective, randomized, controlled trial. AB - PURPOSE: To compare the subjective visual experience of cataract patients during phacoemulsification and intraocular lens implantation under topical anesthesia (TA) vs retrobulbar anesthesia (RA). DESIGN: Prospective, randomized, controlled trial. METHODS: Three hundred six cataract patients eligible for phacoemulsification and intraocular lens implantation were randomized to receive either TA or RA during surgery by one of three surgeons. The surgeons were familiar with both anesthetic techniques and operated on the patients using the technique to which the patients were randomized. A masked interviewer conducted in-person interviews with the patients using a standardized questionnaire about their intraoperative visual experience and their reaction to their visual experience between 30 minutes and 4 hours after the surgery. RESULTS: Two patients (one in each group) had intraoperative posterior capsule rupture and were excluded from analysis. There was no statistically significant difference between TA (n = 154) and RA (n = 150) groups, except that more males compared with females had TA (P = .03). More patients who had TA reported perception of light (P < .001) and colors (P < .001) and a change (either increase or decrease) in light brightness during the course of the surgery (P < .001). There was no statistically significant difference in the proportions of patients who saw movements, flashes, instruments, or the surgeon or medical staff during the operation and who found their visual sensations frightening in the two groups. However, 10.4% in the TA group and 9.3% in the RA group found their visual experience frightening. CONCLUSIONS: More patients undergoing cataract surgery under TA compared with RA reported perception of light, colors, and a change in light brightness. A significant proportion of patients in both groups found the visual experience frightening. PMID- 15531314 TI - Four Japanese male patients with juvenile retinoschisis: only three have mutations in the RS1 gene. AB - PURPOSE: To describe the clinical phenotypes of four unrelated Japanese male patients with juvenile retinoschisis and to investigate occurrences of mutations in the RS1 gene. DESIGN: Observational case series and experimental study. METHODS: Fundus examinations, fluorescein angiography, and single-flash electroretinography (ERG) were carried out. In one patient, optical coherence tomography (OCT) was performed. The coding regions of the RS1 gene that encodes retinoschisin were amplified by polymerase chain reaction (PCR). The PCR products were purified and directly sequenced. RESULTS: The four affected patients showed cystoid- or wheel-like foveal changes with a little or no fluorescein leakage and negative b-wave patterns in both eyes. The OCT images of foveal retinoschisis disclosed that splitting occurs in the putative fibers of Henle. In three patients, we identified three different missense mutations (p.S73P, p.Y89C, p.R209C) in the functionally important discoidin domain of the RS1 gene. The p.S73P mutation has not been previously reported. In contrast, no nucleotide substitutions were detected in the fourth patient whose parents were unrelated and asymptomatic. No other member of this family for three generations has had juvenile retinoschisis. CONCLUSION: Because serine 73 is conserved in the mouse ortholog and other discoidin proteins, the proline 73 allele is therefore very likely to encode a defective retinoschisin. Although the inheritance pattern is uncertain in the patient without the RS1 mutation, the clinical and ERG findings were indistinguishable from those of patients with RS1 mutations. This finding points to the genetic heterogeneity of juvenile retinoschisis. PMID- 15531315 TI - Endophthalmitis after pars plana vitrectomy: Incidence, causative organisms, and visual acuity outcomes. AB - PURPOSE: To investigate the incidence, causative organisms, and visual acuity outcomes associated with endophthalmitis after pars plana vitrectomy. DESIGN: Retrospective, noncomparative, consecutive case series. METHODS: The medical records were reviewed of all patients who developed acute-onset postoperative endophthalmitis (within 6 weeks of surgery) after pars plana vitrectomy at Bascom Palmer Eye Institute between January 1, 1984 and December 31, 2003. RESULTS: During the 20-year study interval, the overall incidence rate of postvitrectomy endophthalmitis was 0.039% (6/15,326). Cultured organisms were Staphylococcus aureus (n = 3), Proteus mirabilus (n = 1), and Staphylococcus epidermidis/Pseudomonas aeruginosa (n = 1); one case was culture-negative. Visual acuity after treatment for endophthalmitis ranged from 2/200 to no light perception, with a final vision of light perception or no light perception in four of six (67%) eyes. CONCLUSION: The incidence of endophthalmitis after pars plana vitrectomy is low but the visual acuity outcomes after treatment are generally poor. PMID- 15531316 TI - Severe pigment epithelial alterations in the treatment area following photodynamic therapy for classic choroidal neovascularization in young females. AB - PURPOSE: Although photodynamic therapy (PDT) is an established treatment for choroidal neovascularization (CNV), the mechanisms are still not completely elucidated. Damage to the retinal pigment epithelium (RPE) was observed following uncomplicated PDT in young patients. DESIGN: Observational case series. METHODS: Four female patients between the age of 26 and 39 years presented with visual loss because of classic CNV. In two 39 years old females the CNV originated secondary to a small chorioretinal scar, in a 26 and a 36-year-old woman the CNV was of idiopathic cause. All patients received standard PDT according to the Treatment of Age-Related Macular Degeneration with Photodynamic Therapy (TAP) Study protocol. RESULTS: One to three months after an uncomplicated PDT with verteporfin, severe pigment epithelial alterations in the treatment area were observed. The neovascular membranes responded favorably to the treatment and demonstrated fibrosis and resolution of leakage. Ophthalmoscopically and angiographically, atrophy of the retinal pigment epithelium was seen precisely delineating the size of the treatment spot used. Vision declined in two patients from 0.3 to 0.1 and 0.15 to 0.1. The two other patients demonstrated an increase of visual acuity from 0.7 to 0.9 and from 0.4 to 0.9. The retinal pigment epithelium alterations did not resolve during follow-up, but remained unchanged in area and intensity. CONCLUSIONS: Characteristic retinal pigment epithelium alterations were observed in young female patients with small classic CNV following PDT. Unusual retinal pigment epithelium damage in young female patients without any associated disease might be related to a possible inherent defect in the RPE or to the hormonal status of this specific patient population. PMID- 15531317 TI - Comparative effect of antiplatelet therapy in retinal vein occlusion evaluated by the particle-counting method using light scattering. AB - PURPOSE: To evaluate platelet aggregation in patients with retinal vein occlusion (RVO) by the light-scattering method and compare the effects of three antiplatelet drugs on aggregate formation. DESIGN: Prospective, nonrandomized, interventional clinical trial. METHODS: (1) Platelet aggregation was measured in 42 patients with untreated branch RVO (BRVO), 26 patients with central RVO (CRVO), and 30 healthy control subjects using a light-scattering platelet aggregometer. Platelet aggregates were classified as small, medium, and large according to light intensity. Total light intensities of each aggregate size were compared between BRVO, CRVO, and control subjects. (2) In 33 patients with RVO, platelet aggregation before and 2 weeks after the administration of ticlopidine, beraprost, or aspirin were compared. RESULTS: (1) There was a statistically significant difference (P = 0.0073) between the control subjects (8.3 x 10(6) a.u.) and CRVO patients (22.9 x 10(6) a.u.) with respect to small aggregates. There was no statistically significant difference with respect to medium and large aggregate formation between the control subjects and either patient group. (2) Compared with aggregates formed in the absence of antiplatelet drugs, ticlopidine significantly inhibited only the formation of small aggregates and beraprost that of all sizes; aspirin did not significantly inhibit the formation of any aggregate sizes. CONCLUSIONS: Increase in small platelet aggregates may be attributable to RVO pathogenesis. Beraprost and ticlopidine appear to inhibit small aggregate formation in RVO patients and may represent effective antiplatelet treatments. The light-scattering method is useful to investigate the pathogenesis of RVO and the effects of antiplatelet drugs. PMID- 15531318 TI - The age-related macular degeneration radiotherapy trial (AMDRT): one year results from a pilot study. AB - PURPOSE: To assess the short-term safety and efficacy of treating subfoveal choroidal neovascularization (CNV) with external beam radiation delivered in 5 x 4 Gy fractions among patients having age-related macular degeneration (AMD). DESIGN: A multicenter prospective randomized controlled pilot study. METHODS: Eighty-eight patients were enrolled through 10 sites and were randomized to radiotherapy (20 Gy delivered in 5 daily fractions of 4 Gy each; 6 MV [N = 41]) or no radiotherapy (sham radiotherapy [N = 22] or observation [N = 25]). Eligibility criteria included visual acuity of at least 20/320 and subfoveal CNV not amenable to treatment. Randomization was stratified by lesion type (new or recurrent CNV) and blood (<50% or >/=50% of the lesion [N = 13]). The primary outcome measure was loss of >/=3 lines of visual acuity. Secondary outcome measures were angiographic response and side effects. RESULTS: At baseline, patient and ocular characteristics were similar between treatment groups. At six months, 9 radiated eyes (26%) and 17 eyes not radiated (49%) lost >/=3 lines of visual acuity (P = .04; stratified chi(2) test). At 12 months, 13 radiated eyes (42%) and 9 observed eyes (49%) lost >/=3 visual acuity lines (P = .60). The radiated group demonstrated smaller lesions and less fibrosis than the nonradiated group (P = .05 and .004, respectively) at 12 months. Radiation induced complications were not observed except for one radiated eye with numerous cotton wool spots and possible radiation retinopathy. CONCLUSIONS: External beam radiation at 5 x 4 Gy may have a modest and short-lived (six month) benefit in preserving visual acuity. PMID- 15531319 TI - Retinal vein cannulation with prolonged infusion of tissue plasminogen activator (t-PA) for the treatment of experimental retinal vein occlusion in dogs. AB - PURPOSE: To evaluate the feasibility, safety, and efficacy of local thrombolytic agents directly injected into occluded retinal veins in an experimental animal model. DESIGN: Experimental animal study. METHODS: This experimental study was performed in two phases. In phase 1, 15 enucleated porcine eyes and 8 in vivo canine eyes were used for the development of the instrumentation and surgical technique required for retinal vein cannulation with prolonged intravascular infusion. In phase 2 of this study, experimental branch retinal vein occlusion was photo-chemically created using an intravenous injection of rose bengal followed by diode laser photocoagulation in eight eyes of eight dogs. Four eyes were treated by retinal vein cannulation and an injection of tissue plasminogen activator (t-PA) using a specifically designed microcatheter, while the remaining four eyes were untreated (control group). The total amount of t-PA injected intravenously ranged from 400 to 1000 mug, infused over a period ranging from 25 to 45 minutes with a mean pressure of 40 psi, resulting in a mean injection flow rate of 0.05 ml/min. The dogs underwent clinical examination, fluorescein angiography, and histologic examination. Main outcome measures were: Achievement of prolonged intravascular infusion of t-PA, changes in fundus appearance, fluorescein angiography, and histology. RESULTS: A microcatheter instrument and a surgical technique for retinal vein cannulation with prolonged intravascular infusion were developed. Cannulation and t-PA infusion for a period of at least 30 minutes was achieved in all four treated eyes with experimental branch retinal vein occlusion. No complications were recorded in all treated eyes. One week and 1 month postoperatively, treated eyes exhibited marked decreases in retinal hemorrhages, retinal vein dilation, and tortuosity, whereas nontreated eyes exhibited persistence of these findings. Fluorescein angiography demonstrated improved circulatory flow in treated relative to nontreated eyes. Histologic analysis confirmed the presence of thrombi in nontreated eyes only. CONCLUSIONS: Retinal vein cannulation with prolonged intravascular injection of t-PA is feasible and safe, and this may offer a new treatment option for retinal vein occlusion. PMID- 15531320 TI - The length of unsutured mucosal margins in external dacryocystorhinostomy. AB - PURPOSE: To compare the extent of mucosal margins remaining unsutured at the end of external dacryocystorhinostomy (DCR) using single or double flap techniques and to evaluate the influence on outcome. DESIGN: Prospective, consecutive case series. METHODS: The number and length of mucosal margins remaining unsutured at the end of external DCR were evaluated in 64 patients. Mucosal anastomosis was performed using either a conservative (group 1) or extended (group 2) double flap, or a single flap (group 3) dissection. Patients were selected for group 1 if the regional anatomy permitted the desired dissection. All other patients were randomized to group 2 or 3. The mean length of unsutured mucosal margins was calculated for each group. Subjects were followed up to 12 months, with final success rate recorded. RESULTS: In groups 1, 2, and 3 respectively: 11, 24 and 29 patients were recruited; four, 12, and 10 mucosal margins remained unsutured; and the mean length of unsutured mucosal margins was 16.36 mm (SD = 2.8), 70.66 mm (SD = 9.3), and 62.00 mm (SD = 6.0). The mean length of unsutured mucosal margins was significantly different, although groups 1 and 2 collectively did not have a significantly different mean length of unsutured mucosal margin from group 3. Mean follow-up was 11 months (SD = 2.2), 63 patients (98.43%) had a successful outcome with no significant difference among groups (P = .429). CONCLUSION: Different patterns of mucosal dissection in external DCR create a different number and extent of unsutured mucosal margins which do not appear to adversely affect the success rate of external DCR. PMID- 15531321 TI - Optical coherence tomography comes of age. PMID- 15531323 TI - Phakic IOLs: a photo finish? PMID- 15531322 TI - Too thick for your drugs. PMID- 15531324 TI - Imaging for neuro-ophthalmic and orbital disease. AB - PURPOSE: To provide an update on imaging of the brain and orbit for ophthalmologists. DESIGN: Literature review. METHODS: A systematic English language medline search and summary of recent literature on imaging of brain and orbit was performed. RESULTS: Computed tomography and magnetic resonance (MR) scanning are the mainstays for the evaluation of most disorders involving the brain and orbit. Computed tomography angiography and magnetic resonance angiography are relatively newer applications that are useful for the evaluation of arterial and venous disorders. Special sequences such as fat suppression and fluid attenuation inversion recovery are useful techniques for specific ophthalmic indications. Diffusion weighted imaging and perfusion-weighted imaging are improving the evaluation of acute stroke. Functional MRI, positron emission tomography scanning and single photon emission computed tomography may provide useful information regarding brain or tumor metabolism. Magnetic resonance spectroscopy has expanded our knowledge of brain function. Newer imaging studies have improved our diagnostic abilities on many fronts, including new sequences, new applications of imaging studies, and functional imaging of brain. CONCLUSION: New imaging techniques for brain and orbit have an increased potential for improving diagnostic yield. Accurate and timely communication with the neuroradiologist can optimize the prescription and interpretation of imaging in ophthalmology. PMID- 15531325 TI - Attrition from ophthalmology residency programs. AB - PURPOSE: To investigate the incidence of and reasons for voluntary resident attrition from ophthalmology training programs. DESIGN: Retrospective survey. METHODS: A survey was mailed to residency program directors of the 121 Accreditation Council for Graduate Medical Education-accredited ophthalmology residency programs asking them to report the number of residents who withdrew from training during 2001 to 2002 and the reasons for withdrawal. RESULTS: Of 102 responding programs, 13 (12.7%) had one resident withdraw from training in the academic year 2001 to 2002. These 13 residents represented 1.1% of all ophthalmology residents in training at the responding institutions during that academic year. The most frequent reason for withdrawing was to enter another medical specialty. CONCLUSIONS: Voluntary resident attrition from ophthalmology training programs is uncommon. The 1.1% attrition rate in this study is the lowest among published reports of attrition from other specialties. PMID- 15531326 TI - Diagnosis of metastatic carcinoma to the cavernous sinus by computed tomograpy guided fine-needle aspiration. AB - PURPOSE: To describe a case of metastatic lung carcinoma to the cavernous sinus and orbital apex diagnosed by fine-needle aspiration guided by computed tomography. DESIGN: Case report. METHOD: A 52-year-old man who presented with right-sided ptosis and ophthalmoplegia was found to have an enhancing mass of the right orbital apex and cavernous sinus extending into the temporal fossa on magnetic resonance imaging. Chest computed tomography revealed hilar adenopathy and a lesion of the right lower lobe of the lung. Bronchial washings and transbronchial needle biopsies of the lung were nondiagnostic. Computed tomography-guided fine-needle aspiration of the temporal fossa portion of the mass was performed. RESULT: Biopsy of the mass showed malignant cells consistent with metastatic non-small cell lung carcinoma. CONCLUSION: Computed tomography guided fine-needle aspiration can be useful in the diagnosis and management of some masses involving the cavernous sinus. PMID- 15531327 TI - Modified sutureless sclerotomies for pars plana vitrectomy. AB - PURPOSE: To describe a modified technique for performing sutureless sclerotomies for pars plana vitrectomy and to examine its efficacy and safety. DESIGN: Retrospective, interventional case series. METHODS: Self-sealing sclerotomies were created in 35 consecutive cases by oblique penetration of the sclera with a 19G MVR blade. RESULTS: All but two (2.9%) of the sclerotomies were watertight without suturing at the end of surgery. A conjunctival bleb was observed during the first postoperative week in one case, but it resolved without intervention. CONCLUSIONS: Construction of sutureless sclerotomies by this technique is simpler and more rapid compared with previously described techniques. The resulting sclerotomies are more resistant to stretching and tearing of the tunnel's roof, they rarely need suturing, and insertion of instruments is facilitated. PMID- 15531328 TI - Intraoperative breakage of a 25-gauge vitreous cutter. AB - PURPOSE: To report breakage of a 25-gauge vitreous cutter during vitreous surgery. DESIGN: Interventional case report. METHODS: A 60-year-old woman was referred for management of an epiretinal membrane at the macula. Visual acuity was 20/100 in the affected left eye. Vitreous surgery using a 25-gauge vitrectomy system was carried out with a combination of conventional cataract surgery. RESULTS: The vitreous cutter was lodged within the sclerotomy cannula after peripheral vitrectomy and was pulled together with the cannula. The cannula was reinserted by trocar, but as the floating peeled epiretinal membrane was dissected with the vitreous cutter, the tip of the cutter was broken and was aspirated with the membrane. Stereoscopic microscopy and scanning electron microscopy demonstrated that the edge that had broken at the cutter port was smooth. CONCLUSION: Although 25-gauge instruments remain useful, care should be taken against rare surgical complications related to their fragility. PMID- 15531329 TI - An explanation of transient visual loss associated with leaking filtering bleb. AB - PURPOSE: To report a presumed cause for the transient visual loss noted by a patient with a leaking bleb following trabeculectomy. DESIGN: Case report. METHODS: Description of an otherwise healthy woman who developed delayed-onset transient visual loss associated with a leaking filtering bleb. RESULTS: A 62 year-old woman who underwent trabeculectomy with mitomycin C in her right eye 5 years earlier (Feb 23, 1999) presented with a 3-month history of transient decreased vision. The apparent cause was a periodic gush of aqueous flowing across the surface of the cornea. CONCLUSIONS: One cause of bleb-associated transient visual loss can be aqueous leaking from the bleb on to the anterior surface of the cornea, disturbing vision as it flows past the visual axis. This is temporarily relieved by blinking. PMID- 15531330 TI - Twice-yearly exams unnecessary for patients taking quetiapine. AB - PURPOSE: To determine the necessity for the heightened level of monitoring for cataract development in patients taking quetiapine (Seroquel, AstraZeneca Pharmaceuticals, Wilmington, Delaware) dictated by the Physicians' Desk Reference. Also, to explore the possibility of cataractogenesis because of quetiapine therapy. DESIGN: Observational case series. METHODS: Data were garnered from a series of 80 case reports collected at the National Registry of Drug-Induced Ocular Side Effects (Portland, Oregon). The World Health Organization (WHO) causality assessment guidelines were used to assess the relationship between quetiapine and cataractogenesis. RESULTS: There were 34 reports of cataracts associated with quetiapine therapy. Average age was 44 years with 23 females and 11 males studied. Average duration of therapy was 29.3 weeks on standard doses. CONCLUSIONS: Cataractogenesis secondary to quetiapine is "unlikely" by WHO guidelines. This probably makes it unnecessary to require biannual ophthalmic examinations as routine eye examinations are sufficient to screen for this condition. PMID- 15531331 TI - Visibility of lamina cribrosa pores and open-angle glaucoma. AB - PURPOSE: To determine prevalence and associations of visible lamina cribrosa pores in an older population and assess whether this feature is associated with open-angle glaucoma (OAG). DESIGN: Case-control study nested within a large, population-based cross-sectional study. METHODS: The Blue Mountains Eye Study examined 3654 persons (82.4% of permanent residents of a defined area) aged over 48 years. Stereo optic disk photographs were taken and graded reliably in a masked fashion. RESULTS: Lamina pore visibility was associated with larger vertical cup-disk ratio and optic disk diameter, hyperopic refraction, and male gender. Pores were slightly less visible with increasing age. They were visible in 70.8% of subjects with open-angle glaucoma but in only 29.3% of normals. This strong association disappeared after adjusting for other variables. CONCLUSION: Lamina cribrosa pores are commonly visible in glaucoma subjects and less commonly in normals. This association, however, is almost entirely because of an increased visibility associated with larger vertical cup-disk ratio and optic disk size. PMID- 15531332 TI - Eyelid sparganosis. AB - PURPOSE: To report a case of sparganosis in the muscle layer of the eyelid. DESIGN: Observational case report. METHODS: A 67-year-old man with migratory painful swelling on the eyelid that was unresponsive to medications was evaluated and treated surgically. RESULTS: Computed tomography showed a 1.5 x 1.5-cm sized, thick-walled lesion and ultrasound biomicroscopy showed hypoechoic tubular and cystic lesions. During surgery, a white, thread-like plerocercoid, 7 cm in length was detected in the orbicularis muscle of the lower eyelid. Histopathologic examination demonstrated the characteristic feature of the sparganum larva and foreign body granulomatous reaction. Serodiagnosis by enzyme-linked immunosorbent assay was positive. Three months postoperatively, the lesion resolved completely. CONCLUSION: Although rare, sparganosis should be suspected in a moving eyelid mass unresponsive to the medical treatment. PMID- 15531333 TI - Facial and ocular injuries associated with methamphetamine production accidents. AB - PURPOSE: To evaluate the relative contribution of methamphetamine production related accidents to facial and ocular injuries admitted to a regional burn unit. DESIGN: Retrospective observational case series. METHODS: Chart review of all patients admitted to the Burn Unit at the University of Iowa Hospitals and Clinics (UIHC) over a 16-month interval. RESULTS: Methamphetamine related accidents were responsible for thirteen (9.8%) of 132 patients admitted with facial burns to UIHC during the study period. All victims were male. In addition to facial burns, eight patients (14 eyes) suffered chemical or thermal burns of the ocular surface of Hughes grade I or worse. One patient required full thickness skins grafts for cicatricial ectropion of all four eyelids. CONCLUSION: In endemic areas such as Iowa, methamphetamine production accidents contribute to as many as 10% of admissions to burn care units for facial and ocular injuries. PMID- 15531335 TI - Propionibacterium acnes endophthalmitis with bacterial sequestration in a Molteno's implant after cataract extraction. AB - PURPOSE: To report a case of Propionibacterium acnes endophthalmitis following uncomplicated cataract extraction with bacterial sequestration in a preexisting Molteno's drainage implant. DESIGN: Interventional case report. METHODS: A 7-year old girl with congenital glaucoma and a preexisting Molteno's drainage implant developed anterior nongranulomatous uveitis 4 months following cataract surgery. P. acnes endophthalmitis was diagnosed by polymerase chain reaction, Southern blot, and electron microscopy. RESULTS: Extraction of the Molteno's implant was required to control the persistent intraocular inflammation and to convert the results of polymerase chain reaction and Southern blot testing of aqueous sample for P. acnes from positive to negative. CONCLUSION: P. acnes may be sequestered in glaucoma implants, potentially requiring implant removal to treat cases of P. acnes endophthalmitis. PMID- 15531334 TI - Novel 615delC mutation in the CRX gene in a Japanese family with cone-rod dystrophy. AB - PURPOSE: To characterize the clinical features of a Japanese family with cone-rod dystrophy associated with a novel 615delC mutation in the cone-rod homeobox (CRX) gene. DESIGN: Case reports and results of DNA analysis. METHODS: Mutational screening by direct sequencing was performed for the three exons in the CRX gene. The clinical features were evaluated by visual acuity measurements, electroretinography, and kinetic visual field testing. RESULTS: A 615delC mutation in the CRX gene was identified and found to cosegregate with cone-rod dystrophy. The ophthalmic findings included cone-rod dystrophy with negative-type electroretinograms (ERGs) and a rapid progression after the age of 40 years. CONCLUSION: These findings indicate that the 615delC mutation causes cone-rod dystrophy with a negative-type ERG. The genotype-phenotype correlation in the CRX gene in our patient and others reported in the literature suggest that the negative-type ERG might be a good sign for having a mutation in the CRX gene. PMID- 15531336 TI - Optic atrophy and macular degeneration as initial presentations of subacute sclerosing panencephalitis. AB - PURPOSE: To report a case of subacute sclerosing panencephalitis in which the initial clinical presentations were optic atrophy and macular degeneration. DESIGN: Observational case report. METHODS: A 9-year-old boy with visual loss attributable to optic atrophy and macular degeneration had a diagnostic examination. RESULTS: The visual acuity levels of both eyes were light perception at presentation. Fundus examination revealed bilateral optic atrophy and macular degenerative changes including retinal pigment epithelial atrophy, macular scarring, and epiretinal membrane formation. The antimeasles IgG titers were high both in serum and cerebrospinal fluid samples. Neurologic symptoms of myoclonus, ataxia, motor, and mental disabilities occurred 4 months after the onset of first visual disturbances and 2 weeks after the patient applied to our clinic. CONCLUSION: Children with optic atrophy and macular degenerative lesions should be examined for sclerosing panencephalitis, even if the classical neurologic symptoms of this disease are not present. PMID- 15531337 TI - High incidence of orbital malignant lymphoma in Japanese patients. AB - PURPOSE: To review 244 cases of orbital tumors to determine pathologic findings in Japanese patients. DESIGN: Observational case series. METHODS: We studied the pathology and origin of tumors in the orbit in 244 consecutive Japanese patients with orbital tumors at our institution from 1981 through 2002 (age 0 to 90 years, mean, 48.7 years; 114 men, 130 women). RESULTS: The most common tumors were lymphoproliferative diseases (n = 114, 42.5%), including malignant lymphoma (n = 59, 24.1%) and reactive lymphoid hyperplasia (n = 45, 18.4%), pleomorphic adenoma (n = 21, 8.6%), and cavernous hemangioma (n = 18, 7.4%). CONCLUSION: The incidence of lymphoproliferative diseases, especially malignant lymphoma, was very high in Japanese patients. PMID- 15531338 TI - Sticky eyelid syndrome. AB - PURPOSE: To report on patients seen with an unusual condition affecting the eyelids. The syndrome manifests as a temporary adhesion which forms between the upper and lower eyelid associated with laxity of the lower lid retractors. This results in a peculiar closure of the lids. DESIGN: Retrospective case reports. METHODS: Charts of affected patients were reviewed for their clinical histories, examination findings, external photographs, and the results of treatment. RESULTS: Ten consecutive patients with the associated findings were reviewed. All cases were unilateral. Seven patients were Asian and three were Caucasian. Nine patients were symptomatic; of these, all were treated conservatively except for one who requested surgery. Two cases are described and photographs are shown. CONCLUSION: Lower eyelid retractor laxity combined with a temporary adhesion between the upper and lower lid results in the clinical findings of Sticky Eyelid Syndrome. PMID- 15531339 TI - Photodynamic therapy and intravitreal triamcinolone for a subretinal neovascularization in bilateral idiopathic juxtafoveal telangiectasis. AB - PURPOSE: To report combined treatment of photodynamic therapy (PDT) with verteporfin and intravitreal triamcinolone acetonide injection for subfoveal neovascularization secondary to bilateral idiopathic juxtafoveal telangiectasis. DESIGN: Interventional case report. METHOD: A patient with a subfoveal neovascularization secondary to bilateral idiopathic juxtafoveal telangiectasis was treated with PDT plus an intravitreal injection of 4 mg of triamcinolone acetonide. RESULTS: Leakage in the late-phase of fluorescein angiography resolved with attenuation of telangiectatic vessels and improvement in visual acuity from 20/200 to 20/50. At 9 months post-treatment, recurrent leakage was treated with repeat PDT and intravitreal triamcinolone. One year after initial presentation, visual acuity was 20/60 with no leakage on fluorescein. CONCLUSION: Combined treatment with PDT and intravitreal triamcinolone acetonide resulted in regression of a subfoveal neovascular membrane and improvement in visual acuity during the course of follow-up. PMID- 15531340 TI - Indocyanine green-assisted ILM peeling in macular hole surgery revisited. AB - PURPOSE: To report visual outcomes after peeling of the internal limiting membrane (ILM) using a brief application of low dose indocyanine green (ICG) for macular hole repair. DESIGN: Prospective, nonrandomized interventional case series. METHODS: A consecutive series of 16 eyes of 14 patients with full thickness macular hole underwent vitrectomy and peeling of the ILM with an intravitreal application of 0.05% ICG for less than 10 seconds. RESULTS: The ILM could be removed uneventfully in 5 of 16 cases after a single ICG application and in 11 cases after a second ICG application. The macular hole was closed in 15 (93.7%) of 16 cases after a single surgery and in one case after a repeat surgery, as evaluated by optical coherence tomography. A significant visual acuity improvement was achieved as the mean logarithm of the minimum angle of resolution was from 0.600 preoperatively (equivalent to approximately 5/20 in the conventional 20 foot numerator form) to 0.213 (12/20) at the end of follow-up (P < .0001). None of the cases developed peripheral visual field defect. Ophthalmoscopies or optic coherence tomographic images did not reveal any disruption at the level of the retinal pigment epithelium. CONCLUSIONS: A brief intravitreal application of low dose ICG may provide a safe ILM peeling in vitreomacular surgeries. PMID- 15531341 TI - Collateral damage in acute zonal occult outer retinopathy. AB - PURPOSE: To evaluate the disease involvement in a patient with acute zonal occult outer retinopathy (AZOOR). DESIGN: Observational case report. METHODS: A patient with acute zonal occult outer retinopathy was imaged with fundus photography, fluorescein and indocyanine green angiography, and autofluorescence photography. RESULTS: There was subtle depigmentation in the central portion of the lesion with a drusen-like deposit at the outer border. Fluorescein angiography showed a transmission defect centrally and a blocking defect at the border where the drusenoid material accumulated. Autofluorescent photography demonstrated that the drusenoid material was intensely autofluorescent, consistent with the presence of lipofuscin, and the central portion of the lesion showed atrophy of the retinal pigment epithelium. Indocyanine green angiography showed atrophy of the choriocapillaris underlying areas of atrophy of the retinal pigment epithelium. CONCLUSIONS: In this case acute zonal occult outer retinopathy caused an area of retinal pigment epithelium cell death with lipofuscin-laden cells at the border of the expanding lesion and associated atrophy of the underlying choriocapillaris. PMID- 15531342 TI - Blindness and rectus muscle damage following spinal surgery. AB - PURPOSE: To report a rare case of blindness and rectus muscle damage from ocular compression after spinal surgery. DESIGN: Observational case report. METHODS: A 16-year-old girl underwent scoliosis surgery in the prone position. After surgery, she complained of visual loss in the right eye. Examination showed no perception of light, periorbital bruising, corneal haze, fixed dilated pupil, and loss of adduction beyond the midline. RESULTS: Orbital computed tomography scan showed a swollen medial rectus with no other orbital pathology. The muscle swelling can be explained by ischemia from compression of the globe against the medial wall of the orbit. Ocular compression also caused retinal and optic nerve ischemia, resulting in loss of vision. Medial rectus function recovered with time, but the patient continues to have no light perception. CONCLUSIONS: Blindness after spinal surgery is a rare but devastating complication, and all precautions to avoid ocular compression must be taken. PMID- 15531343 TI - Lack of apparent short-term benefit of photodynamic therapy in bilateral, acquired, parafoveal telangiectasis without subretinal neovascularization. AB - PURPOSE: To analyze the results of photodynamic therapy (PDT) in bilateral acquired parafoveal telangiectasis. DESIGN: Interventional cases report. METHOD: Retrospective analysis of visual acuity measured with ETDRS charts and of fluorescein angiography findings for two women in whom the eye with lesser visual acuity was treated by PDT for bilateral acquired parafoveal telangiectasis without subretinal neovascularization. RESULTS: Initial visual acuity of the treated eye was 20/63 in both patients and did not change significantly for 3 to 6 months after one session of PDT. Fluorescein angiography in both cases and optical coherence tomography in one case also appeared unchanged. No adverse effect has been observed. CONCLUSIONS: Photodynamic therapy was not beneficial for the outcome of these patients with bilateral acquired telangiectasis, because it improved neither their visual acuity nor their macular edema. PMID- 15531344 TI - The pseudopapilledema of neonatal-onset multisystem inflammatory disease. AB - PURPOSE: To report a rare case of neonatal-onset multisystem inflammatory disease with serial photographs to characterize the optic nerve findings. DESIGN: Observational case report. METHODS: A 6-year-old girl with neonatal-onset multisystem inflammatory disease, who had received systemic corticosteroid therapy for 5 years, had bilateral fibrillar opacities that surrounded the optic disks and extended into the peripapillary nerve fiber layer and vessels. A magnetic resonance imaging examination and lumbar puncture revealed elevated intracranial pressure. RESULTS: The elevated intracranial pressure returned to normal following a corticosteroid taper. Optic disk photographs, taken 4 years earlier, were subsequently obtained. The optic disk appearance had remained unchanged over the 4-year period, consistent with a pseudopapilledema. CONCLUSION: The optic disk appearance is not consistent with papilledema from increased intracranial pressure. The optic disk findings, in conjunction with the underlying inflammatory syndrome, suggest an infiltrative etiology for the atypical optic nerve findings in neonatal-onset multisystem inflammatory disease. PMID- 15531345 TI - Frequency doubling technology perimetry abnormalities as predictors of glaucomatous visual field loss. PMID- 15531347 TI - Age-related changes in corneal and ocular higher-order aberrations. PMID- 15531349 TI - Long-term outcome after discontinuation of benzodiazepines for insomnia: a survival analysis of relapse. AB - Discontinuation of benzodiazepine (BZD) treatment for insomnia can be a difficult task. Cognitive-behavior therapy (CBT) for insomnia, combined with a supervised medication taper, can facilitate withdrawal but there is limited evidence on long term outcome after discontinuation. The objective of this study was to examine medication-free survival time and predictors of relapse (i.e., resumed BZD hypnotics) over a 2-year period in 47 older adults (mean age 62.1 years) with persistent insomnia and prolonged BZD use (average duration of 18.9 years), who had successfully discontinued BZD following CBT for insomnia, a supervised medication taper program, or a combined approach. The Kaplan-Meier product-limit method was used to estimate survival time, defined as time between end-of treatment and relapse or end of follow-up. By the end of the 24-month follow-up, 42.6% of the samples had resumed BZD use. Participants in the Combined (33.3%) and Taper (30.8%) groups relapsed significantly less than their counterparts from the CBT group (69.2%). Survival rates at 3 months were 61.5% (CBT), 100% (Taper), and 80.9% (Combined). At 12 months, they were 38.5%, 83.3%, and 70.8%, respectively; and, at 24 months, they were 28.9%, 64.8% and 64.9%, respectively. Mean survival time was significantly longer for both the Taper (18.6 months, SE = 2.1) and Combined groups (12.6 months, SE = 1.4), relative to the CBT group (8.5 months, SE = 1.8). Significant predictors of relapse included treatment condition, end of treatment insomnia severity, and psychological distress. In conclusion, there is a substantial relapse rate following BZD discontinuation among prolonged users. CBT booster sessions might enhance compliance with CBT and prove useful in preventing relapse. PMID- 15531350 TI - Memory and metamemory in obsessive-compulsive disorder. AB - Studies on the link between checking and memory problems have produced equivocal results regarding a general memory deficit in patients with obsessive-compulsive disorder and subclinical checkers. However, there is clear and consistent evidence that patients with obsessive-compulsive disorder (OCD) show lack of confidence in their memory performance. The purpose of the present study was to investigate memory and metamemory performance (feeling-of-knowing judgments) for neutral and threat-related material in three groups: OCD patients (OCs), subclinical checkers (SCs), and normal controls (NCs). Participants studied a list of neutral and threat word pairs. After an initial cued-recall test, they provided feeling-of-knowing (FOK) judgments for unrecalled word pairs, followed by a recognition test. The results showed that OCs but not SCs were impaired in both recall and recognition compared to NCs. OCs were also less confident about their future memory performance than the other two groups, as reflected in their lower FOK ratings. Moreover, FOK judgments of the OCs were not reliable predictors of their recognition performance. Finally, neither OCs nor SCs showed any evidence of memory bias for threat-relevant information. The results support the idea of a general memory and a metamemory deficit in OCs. PMID- 15531351 TI - Thought-action fusion and its relationship to schizotypy and OCD symptoms. AB - Thought-action fusion (TAF) is a cognitive bias that has been linked to obsessive compulsive disorder (OCD). Preliminary evidence suggests schizotypal traits may be associated with some types of OCD obsessions but not others. We examined the relationship between each of the two major types of TAF (i.e., likelihood and moral), schizotypal traits, and OCD symptoms in 969 nonclinical undergraduate students. We hypothesized that likelihood TAF would be associated with schizotypal traits; whereas moral TAF would not. Consistent with prediction, schizotypal-magical thinking was significantly associated with likelihood TAF even after controlling for the effects of OCD symptoms, general anxiety, and depression. Moreover, the relationship between likelihood TAF and OCD symptoms was significantly attenuated after controlling for schizotypal traits. In contrast, moral TAF demonstrated negligible association with OCD symptoms, depression, or schizotypal traits. These findings provide preliminary support for the linkage between likelihood TAF and schizotypal traits. PMID- 15531352 TI - Reliability and validity of the Padua Inventory in an Iranian population. AB - In order to study the reliability and validity of the Padua Inventory (PI), its items were translated into Persian and then back-translated into English as suggested by Brislin, Lonner, Thorndike, Cross-cultural research methods, Wiley, New York, 1973. Using the translated version of the PI, two studies were carried out. The first study included 219 healthy participants selected from Shiraz city in Iran to examine the reliability and factorial structure of the PI via test retest, internal consistency and principal component analysis methods. The results of this study showed good reliability for the PI and confirmed its factorial structure reported by previous studies. The second study included four groups of participants as follows: patients with obsessive-compulsive disorder (OCD), patients with anxiety disorders, patients with depressive disorder and normal individuals. The PI scores were able to differentiate between OCD patients and normal individuals, but failed to differentiate between neurotic and OCD patients. Two subscales of PI-R version of the PI (precision and washing) differentiated depressive and OCD patients. PMID- 15531353 TI - Is the concept of "repression" useful for the understanding chronic PTSD? AB - Theories concerning the value of avoiding versus attending to trauma-related thoughts provide mixed support for specific coping strategies such as repression. The goal of this study is to examine the usefulness of the concept of repression in understanding chronic Posttraumatic Stress Disorder (PTSD). One hundred and fifty individuals who had been in a motor vehicle accident were included. Participants were classified into four groups (repressors, low anxious, high anxious, and defensively high anxious) based on methodology introduced by Weinberger et al. [J. Abnormal Psychol. 88 (1979) 369]. These four groups were compared on measures of PTSD symptomatology, anxiety, depression, and where appropriate, perceived pain and disability. Results revealed a fairly consistent pattern of group differences such that repressors reported fewer PTSD symptoms, fewer additional anxiety disorders, less depression, and less physical disability due to pain relative to the high anxious and defensively high anxious groups. Regression analyses examining the separate and interactive effects of anxiety and social desirability to predict PTSD symptomatology showed that the majority of the variance was explained by anxiety. In many respects, these data suggest that repression may not be a useful concept for understanding chronic PTSD. PMID- 15531354 TI - A mood-as-input account of perseverative checking: the relationship between stop rules, mood and confidence in having checked successfully. AB - The present paper reports the results of two experiments designed to test predictions from the mood-as-input account of perseverative checking. Using an analogue checking task, both experiments showed that perseveration, as indicted by a range of measures relevant to compulsive checking, was affected by the configuration of the stop rule for the task and mood at the outset of checking. Perseveration was most significant in the condition that most closely resembled the characteristics of obsessive-compulsive checkers (negative mood combined with a stop rule that specifies that the task should be done as thoroughly as possible -namely, an 'as many as can' stop rule). The studies also indicated that confidence at having completed the checking task successfully was (1) significantly related to the use of 'as many as can' stop rules at the outset of checking, (2) mood ratings at the end of checking, and (3) checking perseveration generally. These findings provide support for a mood-as-input explanation of perseverative psychopathologies such as compulsive checking, and begin to cast some light on how anxiety-reduction and 'confidence' models of compulsive checking, might be explained within broader mechanisms of perseveration. PMID- 15531355 TI - Thought suppression and its effects on thought frequency, appraisal and mood state in individuals with obsessive-compulsive disorder. AB - Leading models of obsessive-compulsive disorder (OCD) implicate thought suppression as a key factor in the escalation and persistence of the disorder. This experiment examined the effects of suppression on the frequency of obsessional thoughts in 50 individuals with a primary diagnosis of OCD, and also investigated the effects of participants' appraisals regarding their failures in thought control on distress about intrusive thoughts and on mood. Participants' most upsetting obsessional thought was primed and they then monitored its occurrence over two 4-min intervals. In the first interval, half of the participants were instructed to suppress their obsessional thought and half were instructed not to suppress any thoughts. In the second interval, all participants were given the 'Do Not Suppress' instructions. Participants rated their suppression effort and discomfort over thought occurrences after each interval and recorded their appraisals of thought recurrences during the first interval. Mood state was then reassessed. No ironic effect of suppression on frequency was noted. However, negative appraisals of thought recurrences were associated with more discomfort over thought occurrences and more negative mood at Time 2. These findings suggest that interpretations regarding failures in thought control may be of central importance in understanding obsessional problems. PMID- 15531356 TI - Recognition bias for critical faces in social phobia: a replication and extension. AB - Studies using linguistic stimuli have provided little support for explicit memory biases among individuals with social phobia (SP). However, using facial stimuli rated on their criticalness, Lundh and Ost (1996) found that individuals with SP recognized more critical than accepting faces, whereas non-anxious controls tended to show the opposite pattern. Since the publication of Lundh and Ost's findings, additional studies using a variety of facial stimuli have produced inconsistent findings (J. Anxiety Disord. 14 (2000) 501; Behav. Res. Ther. 39 (2001) 967). Unfortunately, these inconsistencies are difficult to reconcile given great variation in methods and stimuli. Therefore, we designed a study to replicate and extend the work of Lundh and Ost (Behav. Res. Ther. 34 (1996) 787). Similar to Lundh and Ost, individuals with SP identified a significantly higher proportion of old critical faces as old than did non-anxious controls. Further, extending the work of Lundh and Ost, signal detection analyses revealed group differences on response bias according to face type. Specifically, controls showed a response bias towards indicating that accepting faces were previously seen, whereas individuals with SP did not. Finally, signal detection analyses failed to reveal group differences in the accuracy of memory. PMID- 15531357 TI - Mental pollution: feelings of dirtiness without physical contact. AB - An experiment was carried out in order to test the hypothesis that feelings of mental pollution can be induced without physical contact. A sample of 121 female university undergraduates were asked to imagine experiencing a non-consensual kiss at a party, as described on an audiotape, or a consensual kiss described on a comparable audiotape. The manipulation succeeded and participants in the non consensual kiss condition reported significant feelings of mental pollution, negative emotions and cognitions, as well as the urge to wash. Further, eight participants in the non-consensual kiss condition engaged in washing/rinsing behaviour to counteract feelings of mental pollution. The results are consistent with the hypothesis and with reports from an earlier study of victims of sexual assault, a majority of whom described feelings of mental pollution post-assault. The results are also compatible with case descriptions of the onset of mental pollution and OCD subsequent to sexual trauma. Some possible implications of the results, clinical and theoretical are adumbrated. PMID- 15531358 TI - Social anxiety in children: social skills deficit, or cognitive distortion? AB - BACKGROUND: Treatments for childhood social anxiety have traditionally employed social skills training, based on the assumption that effected children have social skills deficits. Recent conceptualisations of social anxiety in adults have questioned this assumption, and have suggested that socially anxious individuals merely believe that they have skill deficits. A recent study using children provided preliminary confirmation of this for younger populations, and also suggested that beliefs about appearing nervous are of particular importance. METHODS: Two groups of children, aged 10-11 years (analogue high social anxiety/low social anxiety), participated in a conversation with an unfamiliar adult. They then rated their performance in a number of domains, after which independent observers also rated their performances. RESULTS: Independent observers were unable to distinguish between the low and high social anxiety groups. However, high socially anxious children rated themselves as appearing significantly less skilled than their low socially anxious counterparts. Notably, high socially anxious children rated themselves particularly poorly in terms of how nervous they looked. CONCLUSIONS: Socially anxious children may not necessarily display social skill deficits. However, they may believe that they appear nervous during social encounters. Clinicians should consider using CBT techniques to address these concerns, rather than relying on social skill remediation. PMID- 15531359 TI - Posttraumatic stress and depressive symptoms in a college population one year after the September 11 attacks: the effect of proximity. AB - As a follow-up to our earlier report [Behav. Res. Ther., in press] on the level of posttraumatic stress symptoms (PTSS), depressive symptoms, and frequency of diagnoses of probable posttraumatic stress disorder (PTSD) among college students at three public universities (Albany, NY, Augusta, GA, and Fargo, ND) resulting from the September 11, 2001. Terrorist attacks, we surveyed comparable groups of students (total, n = 1313) from these three institutions in the weeks following the first anniversary (2002) of the attacks. We found proximity effects (Albany higher than Augusta which was higher than Fargo) for PTSS and depressive symptoms but not for frequency of diagnoses of probable PTSD. Within the Albany site data, proximity of county of residence to New York City (NYC) also showed a proximity effect on PTSS. Although depressive symptoms were significantly different in 2002 versus 2001, the arithmetic differences in PTSS or in frequency of diagnoses of probable PTSD were not significant. The September 11 attacks continued to exert a psychic toll on college students even a year later. PMID- 15531360 TI - Early development of the cranial sensory nervous system: from a common field to individual placodes. AB - Sensory placodes are unique columnar epithelia with neurogenic potential that develop in the vertebrate head ectoderm next to the neural tube. They contribute to the paired sensory organs and the cranial sensory ganglia generating a wide variety of cell types ranging from lens fibres to sensory receptor cells and neurons. Although progress has been made in recent years to identify the molecular players that mediate placode specification, induction and patterning, the processes that initiate placode development are not well understood. One hypothesis suggests that all placode precursors arise from a common territory, the pre-placodal region, which is then subdivided to generate placodes of specific character. This model implies that their induction begins through molecular and cellular mechanisms common to all placodes. Embryological and molecular evidence suggests that placode induction is a multi-step process and that the molecular networks establishing the pre-placodal domain as well as the acquisition of placodal identity are surprisingly similar to those used in Drosophila to specify sensory structures. PMID- 15531361 TI - Signals derived from the underlying mesoderm are dispensable for zebrafish neural crest induction. AB - Signals from the non-neural ectoderm, the neural ectoderm, and the underlying mesoderm have all been implicated in the induction of neural crest. Bone morphogenetic protein (BMP) signaling in particular has an important role in this process; however, it is unclear whether this activity of BMP is due to its effects on patterning the underlying mesoderm, to its ability to establish a competent neural plate boundary zone, or to the direct specification of neural crest at intermediate levels of activity within a BMP gradient. We show neural crest induction occurs in zebrafish in the absence of involuted mesoderm, indicating that this tissue and signals derived from it are dispensable for the formation of neural crest. Dorsal-involuted mesoderm is a major source of secreted BMP antagonists, and the activity of BMP signaling is thought to depend on the presence of the opposing activity of these antagonists. We find that the three BMP antagonists known to be expressed during gastrulation in zebrafish, noggin1, follistatin, and chordin, are dispensable for neural crest induction. These results suggest that mechanisms for restricting the spatio-temporal pattern of BMP expression may compensate for the loss of secreted BMP antagonist activity in establishing dorso-ventral patterning, neural induction, and the neural crest. PMID- 15531362 TI - CD44 expression identifies astrocyte-restricted precursor cells. AB - The precise lineage between neural stem cells and mature astrocytes remains poorly defined. To examine astrocyte development, we have characterized glial precursors from neural tissue derived from early embryonic ages. We show that CD44 identifies an astrocyte-restricted precursor cell (ARP) that is committed to generating astrocytes in vitro and in vivo in both rodent and human tissue. CD44+ cells arise later in development than neuronal-restricted precursors (NRPs) or tripotential glial-restricted precursors (GRPs). ARPs are distinguished from GRP and NRP cells by their antigenic profile and differentiation ability. ARPs can be generated from GRP cells in mass or clonal cultures and in vivo after transplantation, suggesting a sequential differentiation of neuroepithelial stem cells (NEPs) to GRPs to ARPs and then to astrocytes. The properties of ARPs are different from other astrocyte precursors described previously in their expression of CD44 and S-100beta and absence of other lineage markers. Using a CD44 misexpression transgenic mouse model (CNP-CD44 mouse), we show that CD44 overexpression in vivo and in vitro decreases the number of mature glia and increases the number of O4+/GFAP+ cells tenfold. Misexpression of CD44 in culture inhibits oligodendrocytes and arrests cells at the precursor state. In summary, our data provide strong evidence for the existence of a CD44+ ARP in the developing nervous system. PMID- 15531363 TI - Sequences downstream of the bHLH domain of the Xenopus hairy-related transcription factor-1 act as an extended dimerization domain that contributes to the selection of the partners. AB - XHRT1 is a member of the HRT/Hey protein subfamily that are known as Notch effectors. XHRT1 is expressed in the developing floor plate and encodes a basic helix-loop-helix (bHLH) transcription repressor. Here, we show that XHRT1 misexpression in the neural plate inhibits differentiation of neural precursor cells and thus may be important for floor plate cells to prevent them from adopting a neuronal fate. Deletion analysis indicated that inhibition of differentiation by XHRT1 requires the DNA-binding bHLH motif and either the Orange domain or the C-terminal region. XHRT1 could efficiently homodimerize and heterodimerize with hairy proteins. Among those hairy genes, Xhairy2b shows extensive overlap of expression with XHRT1 in floor plate precursors and may be a biologically relevant XHRT1 partner. Dimerization is mediated through both the bHLH and downstream sequences, the Orange domain being particularly important for the efficiency of the interaction. Using chimeric constructs between XHRT1 and the ESR9 bHLH-O protein that does not interact with Xhairy1 and Xhairy2b, we found that both the bHLH domain and downstream sequences of XHRT1 were required for heterodimerization with Xhairy2b, while only the XHRT1 sequences downstream of the Orange domain are required for the interaction with Xhairy1. Together, these results suggest that XHRT1 plays a role in floor plate cell development and highlight the importance of the Orange and downstream sequences in dimerization and in the selection of the bHLH partners. PMID- 15531364 TI - Synergistic roles of BMP15 and GDF9 in the development and function of the oocyte cumulus cell complex in mice: genetic evidence for an oocyte-granulosa cell regulatory loop. AB - Bone morphogenetic protein 15 (BMP15) and growth differentiation factor 9 (GDF9) are oocyte-specific growth factors that appear to play key roles in granulosa cell development and fertility in most mammalian species. We have evaluated the role(s) of these paracrine factors in the development and function of both the cumulus cells and oocytes by assessing cumulus expansion, oocyte maturation, fertilization, and preimplantation embryogenesis in Gdf9+/-Bmp15-/- [hereafter, double mutant (DM)] mice. We found that cumulus expansion, as well as the expression of hyaluronon synthase 2 (Has2) mRNA was impaired in DM oocyte-cumulus cell complexes. This aberrant cumulus expansion was not remedied by coculture with normal wild-type (WT) oocytes, indicating that the development and/or differentiation of cumulus cells in the DM, up to the stage of the preovulatory luteinizing hormone (LH) surge, is impaired. In addition, DM oocytes failed to enable FSH to induce cumulus expansion in WT oocytectomized (OOX) cumulus. Moreover, LH-induced oocyte meiotic resumption was significantly delayed in vivo, and this delayed resumption of meiosis was correlated with the reduced activation of mitogen-activated protein kinase (MAPK) in the cumulus cells, thus suggesting that GDF9 and BMP15 also regulate the function of cumulus cells after the preovulatory LH surge. Although spontaneous in vitro oocyte maturation occurred normally, oocyte fertilization and preimplantation embryogenesis were significantly altered in the DM, suggesting that the full complement of both GDF9 and BMP15 are essential for the development and function of oocytes. Because receptors for GDF9 and BMP15 have not yet been identified in mouse oocytes, the effects of the mutations in the Bmp15 and Gdf9 genes on oocyte development and functions must be produced indirectly by first affecting the granulosa cells and then the oocyte. Therefore, this study provides further evidence for the existence and functioning of an oocyte-granulosa cell regulatory loop. PMID- 15531365 TI - The mouse Forkhead Box m1 transcription factor is essential for hepatoblast mitosis and development of intrahepatic bile ducts and vessels during liver morphogenesis. AB - Conditional deletion of the mouse Forkhead Box (Fox) m1b targeted allele in adult hepatocytes (Foxm1, previously called HFH-11B, Trident, Win, or MPP2) demonstrated that the Foxm1b transcription factor is essential for hepatocyte mitosis during liver regeneration. To determine the role of Foxm1b in liver development, we have generated Foxm1b -/- mice that deleted the Foxm1b exons encoding the winged helix DNA binding and transcriptional activation domains. Here, we show that all of the Foxm1b -/- embryos died in utero by 18.5 days postcoitum (dpc). Embryonic Foxm1b -/- livers displayed a 75% reduction in the number of hepatoblasts, resulting from diminished DNA replication and a failure to enter mitosis causing a polyploid phenotype. Reduced hepatoblast mitosis was associated with decreased protein levels of the Polo-like kinase 1 and Aurora B kinase, which phosphorylate regulatory proteins essential for orchestrating mitosis and cytokinesis. Diminished proliferation of Foxm1b -/- hepatoblasts contributed to abnormal liver development with significant reduction in the number of large hepatic veins compared to embryonic wild-type (WT) liver. Furthermore, embryonic Foxm1b -/- livers did not develop intrahepatic bile ducts, and these presumptive biliary hepatoblasts failed to express either biliary cytokeratins or nuclear levels of hepatocyte nuclear factor 1beta. These results suggest that Foxm1b is critical for hepatoblast precursor cells to differentiate toward biliary epithelial cell lineage. Finally, we used a hepatoblast-specific Cre recombinase transgene to mediate deletion of the Foxm1b fl/fl allele in the developing liver, and these embryos died in utero and exhibited diminished hepatoblast proliferation with similar abnormalities in liver morphogenesis, suggesting that the defect in liver development contributed to embryonic lethality. PMID- 15531366 TI - The Wingless morphogen gradient is established by the cooperative action of Frizzled and Heparan Sulfate Proteoglycan receptors. AB - We have examined the respective contribution of Heparan Sulfate Proteoglycans (HSPGs) and Frizzled (Fz) proteins in the establishment of the Wingless (Wg) morphogen gradient. From the analysis of mutant clones of sulfateless/N deacetylase-sulphotransferase in the wing imaginal disc, we find that lack of Heparan Sulfate (HS) causes a dramatic reduction of both extracellular and intracellular Wg in receiving cells. Our studies, together with others [Kirkpatrick, C.A., Dimitroff, B.D., Rawson, J.M., Selleck, S.B., 2004. Spatial regulation of Wingless morphogen distribution and signalling by Dally-like protein. Dev. Cell (in press)], reveals that the Glypican molecule Dally-like Protein (Dlp) is associated with both negative and positive roles in Wg short- and long-range signaling, respectively. In addition, analyses of the two Fz proteins indicate that the Fz and DFz2 receptors, in addition to transducing the signal, modulate the slope of the Wg gradient by regulating the amount of extracellular Wg. Taken together, our analysis illustrates how the coordinated activities of HSPGs and Fz/DFz2 shape the Wg morphogen gradient. PMID- 15531367 TI - The role of erythropoietin in regulating angiogenesis. AB - Erythropoietin (EPO) is an essential growth factor that regulates erythrocyte production in mammals. In this study, we demonstrate a novel role of EPO in regulating angiogenesis in vivo. Epo and Epo receptor (EpoR) are expressed in the vasculature during embryogenesis. Deletion of Epo or EpoR leads to angiogenic defects starting at E10.5, 2 days before ventricular hypoplasia and 3 days before the onset of the embryonic lethal phenotype. Overall, angiogenesis was severely affected in the mutant embryos: vascular anomalies included decreased complexity of the vessel networks. However, de novo vasculogenesis remained intact, consistent with the differential expression of Epo and EpoR during the early stages of embryonic development. The aforementioned angiogenesis defect can be partially rescued by expressing human EPO during embryogenesis. Moreover, Ang-1 expression is regulated by EPO/EPOR under normoxic conditions. Taken together, our results suggest important roles of EPO and EPOR in angiogenesis. PMID- 15531368 TI - Aym1, a mouse meiotic gene identified by virtue of its ability to activate early meiotic genes in the yeast Saccharomyces cerevisiae. AB - Our understanding of the molecular mechanisms that operate during differentiation of mitotically dividing spermatogonia cells into spermatocytes lags way behind what is known about other differentiating systems. Given the evolutionary conservation of the meiotic process, we screened for mouse proteins that could specifically activate early meiotic promoters in Saccharomyces cerevisiae yeast cells, when fused to the Gal4 activation domain (Gal4AD). Our screen yielded the Aym1 gene that encodes a short peptide of 45 amino acids. We show that a Gal4AD AYM1 fusion protein activates expression of reporter genes through the promoters of the early meiosis-specific genes IME2 and HOP1, and that this activation is dependent on the DNA-binding protein Ume6. Aym1 is transcribed predominantly in mouse primary spermatocytes and in gonads of female embryos undergoing the corresponding meiotic divisions. Aym1 immunolocalized to nuclei of primary spermatocytes and oocytes and to specific type A spermatogonia cells, suggesting it might play a role in the processes leading to meiotic competence. The potential functional relationship between AYM1 and yeast proteins that regulate expression of early meiotic genes is discussed. PMID- 15531369 TI - Conditional deletion of the TGF-beta type II receptor in Col2a expressing cells results in defects in the axial skeleton without alterations in chondrocyte differentiation or embryonic development of long bones. AB - Members of the TGF-beta superfamily are secreted signaling proteins that regulate many aspects of development including growth and differentiation in skeletal tissue. There are three isoforms of TGF-beta that act through the same heteromeric receptor complex. To address the question of the role of TGF-beta signaling in skeletal development, we generated mice with a conditional deletion of the TGF-beta type II receptor gene (Tgfbr2) specifically in Col2a expressing cells using the Cre/lox recombinase system. Alizarin red-/Alcian blue-stained skeletons were prepared from embryos at 17.5, 15.5, and 13.5 days of gestation. Col2acre+/-;TgfbrloxP/loxP and Col2acre-/-;Tgfbr2+/loxP skeletons were compared. Multiple defects were observed in the base of the skull and in the vertebrae. Specifically, the size and spacing of the vertebrae were altered, and defects were detected in the closure of the neural arches. In addition, alterations in transverse processes, costal joints, and zygapophyses were detected. While the vertebral bodies were only moderately affected, the intervertebral discs (IVDs) were either missing or incomplete. Alterations in the vertebrae could be detected as early as E13.5 days. Surprisingly, alterations in length and mineralization of long bones were not detected at E17.5 days. In addition, the expression patterns of markers for chondrocyte differentiation were not altered in vertebrae or long bones suggesting that loss of responsiveness to TGF-beta in chondrocytes does not affect embryonic endochondral bone formation. In contrast, mice that survived postnatally demonstrated alterations in the length of specific bones. Skeletons from Col2acre+/-;Tgfbr2loxP/loxP mice were compared to those from mice null for the TGF-beta2 ligand. The differences observed between these models allow distinctions to be made between the roles of the various isoforms of TGF-beta and the signaling in specific cell types. The data provide information regarding mechanisms of skeletal development and suggest that TGF-beta signaling is a critical component. PMID- 15531370 TI - CYP26A1 and CYP26C1 cooperate in degrading retinoic acid within the equatorial retina during later eye development. AB - In the embryonic mouse retina, retinoic acid (RA) is unevenly distributed along the dorsoventral axis: RA-rich zones in dorsal and ventral retina are separated by a horizontal RA-poor stripe that contains the RA-inactivating enzyme CYP26A1. To explore the developmental role of this arrangement, we studied formation of the retina and its projections in Cyp26a1 null-mutant mice. Expression of several dorsoventral markers was not affected, indicating that CYP26A1 is not required for establishing the dorsoventral retina axis. Analysis of the mutation on a RA reporter mouse background confirmed, as expected, that the RA-poor stripe was missing in the retina and its projections at the time when the optic axons first grow over the diencephalon. A day later, however, a gap appeared both in retina and retinofugal projections. As explanation, we found that CYP26C1, another RA degrading enzyme, had emerged centrally in a narrower domain within the RA-poor stripe. While RA applications increased retinal Cyp26a1 expression, they slightly reduced Cyp26c1. These observations indicate that the two enzymes function independently. The safeguard of the RA-poor stripe by two distinct enzymes during later development points to a role in maturation of a significant functional feature like an area of higher visual acuity that develops at its location. PMID- 15531371 TI - Developmental expression and function of Bmp4 in spermatogenesis and in maintaining epididymal integrity. AB - Bone morphogenetic proteins (BMPs) play essential roles in many aspects of developmental biology. We have previously shown that Bmp7, Bmp8a, and Bmp8b of the 60A class of Bmp genes have additive effects in spermatogenesis and in maintaining the epididymal integrity of the caput and caudal regions. Here we report that Bmp4 of the Dpp class has a unique expression pattern in the developing testis and epididymis. Bmp4 heterozygous males on a largely C57BL/6 background show compromised fertility due to degeneration of germ cells, reduced sperm counts, and decreased sperm motility. More interestingly, some of these males show extensive degeneration of the epididymal epithelium in the corpus region, rather than in the caput and cauda regions as for Bmp7 and Bmp8 mutants. Thus, these genetic data reveal a region-specific requirement of different classes of BMPs for epididymal epithelium to survive and have significant implications on male reproductive health and perhaps birth control. PMID- 15531372 TI - Notch promotes survival of neural precursor cells via mechanisms distinct from those regulating neurogenesis. AB - During development of the mammalian brain, many neural precursor cells (NPCs) undergo apoptosis. The regulation of such cell death, however, is poorly understood. We now show that the survival of mouse embryonic NPCs in vitro was increased by culture at a high cell density and that this effect was attributable to activation of Notch signaling. Expression of an active form of Notch1 thus markedly promoted NPC survival. Hes proteins, key effectors of Notch signaling in inhibition of neurogenesis, were not sufficient for the survival-promoting effect of Notch1. This effect of Notch1 required a region of the protein containing the RAM domain and was accompanied by up-regulation of the anti-apoptotic proteins Bcl-2 and Mcl-1. Moreover, knockdown of these proteins by RNA interference resulted in blockade of the Notch1-induced survival. These results reveal a new function of Notch, the promotion of NPC survival. PMID- 15531373 TI - BMP signaling through ACVRI is required for left-right patterning in the early mouse embryo. AB - Vertebrate organisms are characterized by dorsal-ventral and left-right asymmetry. The process that establishes left-right asymmetry during vertebrate development involves bone morphogenetic protein (BMP)-dependent signaling, but the molecular details of this signaling pathway remain poorly defined. This study tests the role of the BMP type I receptor ACVRI in establishing left-right asymmetry in chimeric mouse embryos. Mouse embryonic stem (ES) cells with a homozygous deletion at Acvr1 were used to generate chimeric embryos. Chimeric embryos were rescued from the gastrulation defect of Acvr1 null embryos but exhibited abnormal heart looping and embryonic turning. High mutant contribution chimeras expressed left-side markers such as nodal bilaterally in the lateral plate mesoderm (LPM), indicating that loss of ACVRI signaling leads to left isomerism. Expression of lefty1 was absent in the midline of chimeric embryos, but shh, a midline marker, was expressed normally, suggesting that, despite formation of midline, its barrier function was abolished. High-contribution chimeras also lacked asymmetric expression of nodal in the node. These data suggest that ACVRI signaling negatively regulates left-side determinants such as nodal and positively regulates lefty1. These functions maintain the midline, restrict expression of left-side markers, and are required for left-right pattern formation during embryogenesis in the mouse. PMID- 15531374 TI - The cfy mutation disrupts cell divisions in a stage-dependent manner in zebrafish embryos. AB - The zebrafish curly fry (cfy) mutation leads to embryonic lethality and abnormal cell divisions starting at 12-14 h postfertilization (hpf) during neural tube formation. The mitotic defect is seen in a variety of tissues including the central nervous system (CNS). In homozygous mutant embryos, mitoses are disorganized with an increase in mitotic figures throughout the developing neural tube. One consequence of aberrant mitoses in cfy embryos is an increase in cell death. Despite this, patterning of the early CNS is relatively unperturbed with distribution of the early, primary neurons indistinguishable from that of wild type embryos. At later stages, however, the number of neurons was dramatically decreased throughout the CNS. The effect on neurons in older cfy embryos but not young ones correlates with the time of birth of neurons: primary neurons are born before the action of the cfy gene and later neurons after. Presumably, death of neuronal progenitors that divide beginning at the neural keel stage or death of their neuronal progeny accounts for the diminution of neurons in older mutant embryos. In addition, oligodendrocytes, which also develop late in the CNS, are greatly reduced in number in cfy embryos due to an apparent decrease in oligodendrocyte precursors. Genetic mosaic analysis demonstrates that the mutant phenotype is cell-autonomous. Furthermore, there are no obvious defects in apical/basal polarity within the neuroepithelium, suggesting that the cfy gene is not critical for epithelial polarity and that polarity defects are unlikely to account for the increased mitotic figures in mutants. These results suggest that the cfy gene regulates mitosis perhaps in a stage-dependent manner in vertebrate embryos. PMID- 15531375 TI - A new origin for the maxillary jaw. AB - One conserved feature of craniofacial development is that the first pharyngeal arch has two components, the maxillary and mandibular, which then form the upper and lower jaws, respectively. However, until now, there have been no tests of whether the maxillary cells originate entirely within the first pharyngeal arch or whether they originate in a separate condensation, cranial to the first arch. We therefore constructed a fate map of the pharyngeal arches and environs with a series of dye injections into stage 13-17 chicken embryos. We found that from the earliest stage examined, the major contribution to the maxillary bud is from post optic mesenchyme with a relatively minor contribution from the maxillo-mandibular cleft. Cells labeled within the first pharyngeal arch contributed exclusively to the mandibular prominence. Gene expression data showed that there were different molecular codes for the cranial and caudal maxillary prominence. Two of the genes examined, Rarbeta (retinoic acid receptor beta) and Bmp4 (bone morphogenetic protein) were expressed in the post-optic mesenchyme and epithelium prior to formation of the maxillary prominence and then were restricted to the cranial half of the maxillary prominence. In order to determine the derivatives of the maxillary prominence, we performed focal injections of CM-DiI into the stage 24 maxillary prominence. Labeled cells contributed to the maxillary, palatine, and jugal bones, but not the other elements of the upper beak, the premaxilla and prenasal cartilage. We also determined that the cranial cells give rise to more distal parts of the upper beak, whereas caudal cells form proximal structures. Grafts of stage 24 maxillary prominences were also analyzed to determine skeletal derivatives and these results concurred with the DiI maps. These early and later fate maps indicate that the maxillary prominence and its skeletal derivatives are not derived from the first pharyngeal arch but rather from a separate maxillary condensation that occurs between the eye and the maxillo-mandibular cleft. These data also suggest that during evolution, recession of the first pharyngeal arch derived palatoquadrate cartilage to a more proximal position gave way to the bony upper jaw of amniotes. PMID- 15531377 TI - Effects of prostanoids on phenylephrine-induced contractions in the mesenteric vascular bed of rats with streptozotocin-induced diabetes mellitus. AB - The main aim of this study was to compare the vascular reactivity of the perfused (Krebs, 4 ml/min) mesenteric vascular bed (MVB) isolated from rats with streptozotocin (STZ)-induced diabetes of 8 weeks duration to that of the MVB from non-diabetic (ND) Wistar rats. There were no differences in basal perfusion pressure between the MVB isolated from STZ and ND rats. The addition of indomethacin to the perfusate increased the basal perfusion pressure in both ND (18.8 +/- 0.7 vs 29.4 +/- 3.7 mmHg, p < 0.05) and STZ rats (18.3 +/- 0.9 vs 27.2 +/- 2.6 mmHg, p < 0.05), suggesting the release of a vasodilator prostaglandin. Remotion of the endothelium did not affect this response, indicating that prostaglandin was released from vascular smooth muscle. The response to phenylephrine was reduced in STZ rats compared to ND rats (2.3 [1.6-3.8] vs 8.3 [3.5-19.4], ED50. [IC 95%]) and was not modified by removal of the endothelium or by perfusion of L-nitro-arginine (50 microM). In contrast, indomethacin was able to reduce the response to phenylephrine in ND but not in STZ rats (2.3 [1.6-3.8] vs 4.7 [3.2-6.0], ED50. [IC 95%], p=0.02), suggesting that the blunted response to phenylephrine observed in STZ was due to the abolition of the release of prostaglandin by vascular smooth muscle. In conclusion, experimental diabetes induction in the rat is followed by a reduction of the contractile effect of phenylephrine due to the lack of release of a vasoconstrictor prostaglandin from vascular smooth muscle. PMID- 15531376 TI - Developmental origins and evolution of jaws: new interpretation of "maxillary" and "mandibular". AB - Cartilage of the vertebrate jaw is derived from cranial neural crest cells that migrate to the first pharyngeal arch and form a dorsal "maxillary" and a ventral "mandibular" condensation. It has been assumed that the former gives rise to palatoquadrate and the latter to Meckel's (mandibular) cartilage. In anamniotes, these condensations were thought to form the framework for the bones of the adult jaw and, in amniotes, appear to prefigure the maxillary and mandibular facial prominences. Here, we directly test the contributions of these neural crest condensations in axolotl and chick embryos, as representatives of anamniote and amniote vertebrate groups, using molecular and morphological markers in combination with vital dye labeling of late-migrating cranial neural crest cells. Surprisingly, we find that both palatoquadrate and Meckel's cartilage derive solely from the ventral "mandibular" condensation. In contrast, the dorsal "maxillary" condensation contributes to trabecular cartilage of the neurocranium and forms part of the frontonasal process but does not contribute to jaw joints as previously assumed. These studies reveal the morphogenetic processes by which cranial neural crest cells within the first arch build the primordia for jaw cartilages and anterior cranium. PMID- 15531378 TI - Diabetes enhances apoptosis induced by cerebral ischemia. AB - The aim of this study is to explore the mechanism by which diabetes exaggerates cerebral stroke and its outcome. Since ischemia can be related to not only necrosis but apoptosis as well, we compared the development of apoptosis in STZ diabetic rats and STZ-diabetic rats subjected to occlusion of the middle cerebral artery (MCA). 24-48 hr following MCA occlusion the animals were killed, the brain removed and prepared for evaluation by several indexes of apoptosis: nucleosomal DNA fragmentation, TUNEL staining, activation of caspase-3 and alteration in the expression of Bax and Bcl2. DNA fragmentation was not detected in the cortex of normal and diabetic animals, but was evident following MCA occlusion in diabetic rats. Bax expression was increased in the cortex of normal rats following MCA occlusion and this expression was further increased in the cortex of MCA occluded diabetic rats. Bcl2 expression was not changed in any of the groups. In the hippocampus, DNA fragmentation was not evident in control rats but was observed in diabetic rats. Ischemic injury did not enhance DNA laddering in diabetic animals. The expression of Bax was increased in diabetic rats but was not increased following MCA occlusion. Bcl2 expression was not changed by ischemia in any of the animal models. These data suggest that diabetes may enhance the development of stroke via increased cortical apoptotic activity but this was not additive in the hippocampus following ischemic injury. PMID- 15531379 TI - Characterization of the human nicotinic acetylcholine receptor subunit alpha (alpha) 9 (CHRNA9) and alpha (alpha) 10 (CHRNA10) in lymphocytes. AB - Though the nicotinic acetylcholine receptor (nAChR) subunits alpha9 and alpha 10 have been thoroughly characterized within hair cells of the organ of Corti in the inner ear, prior studies have shown that they are also expressed in lymphocytes. In this report, we sought to more definitively characterize the nAChR subunits alpha9 and alpha10 within various populations of human lymphocytes. Using a combination of techniques, including RT-PCR, single-cell RT-PCR, Northern and western blot analysis, and immunofluorescence, expression of both alpha9 and alpha 10 was demonstrated in purified populations of T-cells (CD3+, CD4+, CD8+ and the Jurkat, MT2 and CEM T-cell lines) and B-cells (CD19+, CD80+ and EBV immortalized B-cells). Single-lymphocyte recording techniques failed to identify an ionic current in response to applied acetylcholine in either T-cells or B cells. These results clearly demonstrate the presence of these nicotinic receptor subunits within several populations of human lymphocytes, implicating their role in the immune response. However, a lack of demonstrated response to applied acetylcholine using standard single-cell recording techniques suggests a physiology different than that seen in hair cells of the inner ear. PMID- 15531380 TI - Pharmacokinetic and pharmacodynamic interactions between simvastatin and diltiazem in patients with hypercholesterolemia and hypertension. AB - Pharmacokinetic and pharmacodynamic interactions between simvastatin, a 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, and diltiazem, a calcium antagonist, were investigated in 7 male and 4 female patients with hypercholesterolemia and hypertension. The patients were given, for one in a three consecutive 4-week periods, oral simvastatin (5 mg/day), oral simvastatin (5 mg/day) combined with diltiazem (90 mg/day), and then oral diltiazem (90 mg/day), respectively. The area under the plasma concentration versus time curve up to 6 hours post-dose (AUC0-6h) and maximum plasma concentrations (Cmax) of the drugs, serum lipid profiles, blood pressures and liver functions were assessed on the last day of each of the three 4-week periods. After the combined treatment period, Cmax of HMG-CoA reductase inhibitor was elevated from 7.8 +/- 2.6 ng/ml to 15.4 +/- 7.9 ng/ml (P < 0.01) and AUC0-6h from 21.7 +/- 4.9 ng x hr/ml to 43.3 +/- 23.4 ng x hr/ml (P < 0.01), while Cmax of diltiazem was decreased from 74.2 +/- 36.4 ng/ml to 58.6 +/- 18.9 ng/ml (P < 0.05) and its AUC0-6h from 365 +/- 153 ng x hr/ml to 287 +/- 113 ng x hr/ml (P < 0.01). Compared to simvastatin monotherapy, combined treatment further reduced LDL-cholesterol levels by 9%, from 129 +/- 16 mg/dl to 119 +/- 17 mg/dl (P < 0.05). No adverse events were observed throughout the study. These apparent pharmacokinetic interactions, namely the increase of HMG-CoA reductase inhibitor concentration by diltiazem and the decrease of diltiazem concentration by simvastatin, enhance the cholesterol lowering effects of simvastatin during combined treatment. PMID- 15531381 TI - Effects of grapefruit juice on the multidrug transporter P-glycoprotein in the human proximal tubular cell line HK-2. AB - The multidrug transporter MDR-1 P-glycoprotein (Pgp) has been recently pointed out as an important mechanism underlying chemical interaction between drugs and many commonly ingested substances, including grapefruit juice (GFJ). Modulation of intestinal Pgp dependent transport by GFJ may lead to changes in bioavailability of drugs that are substrates of Pgp itself, by affecting their presystemic clearance. Since other cellular sites expressing Pgp and devoted to drug disposition, like kidney proximal tubules, could be involved in these pharmacokinetic interactions, we investigated the effect of GFJ on the expression and activity of Pgp in the human immortalized tubular cell line HK-2. Two flavonoid compounds related to GFJ, kaempferol and naringenin, were also tested for their effects on HK-2 Pgp. HK-2 cells cultured for 4 days in the presence of GFJ, showed a dose-dependent decrease in Pgp immunoblottable amount as well as a decrease in MDR-1 mRNA level, as shown by western blot analysis and RT-PCR, respectively. Both kaempferol and naringenin were also able to significantly decrease Pgp immunoblottable amount. To test whether the downregulation of HK-2 Pgp due to GFJ exposition could influence the cell sensitivity to drugs that are transported by Pgp itself, HK-2 cells precultured with GFJ were exposed to scalar concentrations of Cyclosporin A or Vinblastine and cell viability examined 36 hours later. The cytotoxicity of both drugs was increased. The calcein-AM test in untreated cells showed that GFJ, kaempferol or naringenin inhibited Pgp activity. Downregulation of Pgp as well inhibition of its function by GFJ or its related components in tubular cells could have a role in changing disposition kinetics of some important therapeutic agents. PMID- 15531382 TI - Chronic exposure of cells expressing recombinant GABAA receptors to benzodiazepine antagonist flumazenil enhances the maximum number of benzodiazepine binding sites. AB - The aim of this study was to better understand the mechanisms that underlie adaptive changes in GABAA receptors following their prolonged exposure to drugs. Exposure (48 h) of human embryonic kidney (HEK) 293 cells stably expressing recombinant alpha1beta2gamma2S GABAA receptors to flumazenil (1 or 5 microM) in the presence of GABA (1 microM) enhanced the maximum number (Bmax) of [3H]flunitrazepam binding sites without affecting their affinity (Kd). The flumazenil-induced enhancement in Bmax was not counteracted by diazepam (1 microM). GABA (1 nM-1 mM) enhanced [3H]flunitrazepam binding to membranes obtained from control and flumazenil-pretreated cells in a concentration dependent manner. No significant differences were observed in either the potency (EC50) or efficacy (Emax) of GABA to potentiate [3H]flunitrazepam binding. However, in flumazenil pretreated cells the basal [3H]flunitrazepam and [3H]TBOB binding were markedly enhanced. GABA produced almost complete inhibition of [3H]TBOB binding to membranes obtained from control and flumazenil treated cells. The potencies of GABA to inhibit this binding, as shown by a lack of significant changes in the IC50 values, were not different between vehicle and drug treated cells. The results suggest that chronic exposure of HEK 293 cells stably expressing recombinant alpha1beta2gamma2S GABAA receptors to flumazenil (in the presence of GABA) up-regulates benzodiazepine and convulsant binding sites, but it does not affect the allosteric interactions between these sites and the GABA binding site. Further studies are needed to elucidate these phenomena. PMID- 15531383 TI - Beneficial effect of trimebutine and N-monodesmethyl trimebutine on trinitrobenzene sulfonic acid-induced colitis in rats. AB - The use of local anesthetics, such as lidocaine, has been proposed in the treatment of distal ulcerative colitis. Trimebutine maleate (TMB) displays a local anesthetic activity higher than that of lidocaine in rabbit corneal reflex. TMB and nor-TMB its main metabolite in human show similar affinity to that of bupivacaine toward sodium channel labeled by [3H]batrachotoxin and block sodium currents in sensory neurons from rat dorsal root ganglia. The aim of this study was to evaluate the effects of TMB and nor-TMB in comparison to lidocaine and bupivacaine in a rat model of acute colonic inflammation induced by trinitrobenzene sulfonic acid (TNBS). A single intracolonic instillation of TNBS (50 mg/kg dissolved in ethanol 30%) led to early plasma extravasation then macroscopic damage (hyperemia and necrosis), increased colonic weight and tissular MPO, a marker of neutrophilic infiltration. Local administration of TMB at dose of 3 to 60 mg/kg, 30 min before, 24 and 48 h after colitis induction, significantly reduced the severity of colitis. Nor-TMB (1, 3, 10, 30 mg/kg) as well as lidocaine (1, 3, 10 mg/kg) dose-dependently reduced colitis while bupivacaine at 10 mg/kg did not affect it significantly. In contrast systemic administration of TMB, nor-TMB and lidocaine at 10 mg/kg had no significant effect. Furthermore, local administration of TMB (30 mg/kg) and lidocaine (10 mg/kg) significantly reduced plasmatic extravasation. In conclusion, intracolonic treatment with TMB and nor-TMB improved acute experimental TNBS-induced colitis in rat and these effects could be explained by their local anesthetic activity. PMID- 15531384 TI - Protective effect of rhynchophylline and isorhynchophylline on in vitro ischemia induced neuronal damage in the hippocampus: putative neurotransmitter receptors involved in their action. AB - Rhynchophylline and isorhynchophylline are major tetracyclic oxindole alkaloid components of Uncaira species, which have been long used as medicinal plants. In this study we examined the protective effects of rhynchophylline and isorhynchophylline on in vitro ischemia-induced neuronal damage in the hippocampus and interaction of these alkaloids with neurotransmitter receptors in a receptor expression model of Xenopus oocytes. In vitro ischemia was induced by exposing the hippocampal slices to oxygen- and D-glucose-deprived medium over 8 min. The resultant neuronal damage was elucidated as deterioration of population spike (PS) amplitudes evoked trans-synaptically by electrical stimulation of Schaffer collaterals and recorded in the CA1 area. Rhynchophylline and isorhynchophylline, as well as the N-methyl-D-aspartate (NMDA) antagonist (+/-)-2 amino-5-phosphono-valeric acid (APV), the muscarinic M1 receptor antagonist pirenzepine, and the 5-HT2 receptor antagonist ketanserin, attenuated the in vitro ischemia-induced neuronal damage in a concentration-dependent manner. There was no difference in the extent of protection against the neuronal damage between rhynchophylline and isorhynchophylline treatment. In Xenopus oocytes expressing the rat brain receptors encoded by total RNA, both rhynchophylline and isorhynchophylline reduced muscarinic receptor- and 5-HT2 receptor-mediated current responses in a competitive manner. Together with our previous findings that rhynchophylline and isorhynchophylline have a non-competitive antagonistic effect on the NMDA-type ionotropic glutamate receptors, the present results suggest that these alkaloids exert their protective action against ischemia induced neuronal damage by preventing NMDA, muscarinic M1, and 5-HT2 receptors mediated neurotoxicity during ischemia. PMID- 15531385 TI - Effects of pentadecapeptide BPC157 on regional serotonin synthesis in the rat brain: alpha-methyl-L-tryptophan autoradiographic measurements. AB - A novel pentadecapeptide, BPC157, was recently reported to have a large spectrum of in vivo activities, from anti-ulcer to central action on the brain dopaminergic system. The mechanisms of these actions are not well understood. In this study, the evaluation of the effects of acute and repeated administration of BPC157 on serotonin (5-HT) synthesis in the rat brain is reported. The alpha [14C]methyl-L-tryptophan (alpha-MTrp) autoradiographic method was used to measure regional 5-HT synthesis rates. In the first series of experiments, a single dose treatment of BPC157 (10 microg/kg) administered intraperitoneally 40 min before the alpha-MTrp tracer injection significantly reduced the regional rate of 5-HT synthesis in the dorsal thalamus, hippocampus, lateral geniculate body and hypothalamus. 5-HT synthesis rates in the substantia nigra reticulate and medial anterior olfactory nucleus in BPC157 treated rats were significantly higher than in the control rats. No significant change in the synthesis rate was observed in the raphe nuclei. In the second series of experiments, following a 7-day treatment with BPC157 (10 microg/kg; s.c.), a significant reduction in the 5-HT synthesis rate was observed in the dorsal raphe nucleus, and significant increases were observed in the substantia nigra, lateral caudate, accumbens nucleus and superior olive. This data suggests that BPC157, a gut peptide, influences brain 5-HT synthesis in rats, but we cannot determine, from this data, the mechanism of this action. PMID- 15531386 TI - A new methodology to calculate the environmental protection index (Ep). A case study applied to a company producing composite materials. AB - Environmental indicators can be used as a first stage in progress towards comprehensive environmental impact measures [J. Environ. Manage 65/3 (2002) 285]. In this article, we develop a 'pollutant interaction matrix method' that allows calculation of a global environmental protection index (Ep) in order to verify the eco-compatibility of an industrial activity. Two methods are proposed for the Ep index evaluation (which represents the numerical measure of the environmental sustainability): the Direct Method (Epd) and the Weighted Method (Epw). Both methods need to define, in the whole industrial process, homogeneous sectors (defined as construction sites where activities of the same type are carried out). Furthermore, for each activity a set of parameters (t, duration of pollution effect, P, quantity of pollutant produced,G, hazard of the pollutant) are required to evaluate the relative pollution index Y. All indices calculations were carried out using a set of matrices. The correct use of Ep evaluation provides an improvement in the total environmental performance of companies because it points out possible critical operations in each homogeneous sector which require solutions. The methodology is applied to evaluate the environmental pollution risk of a company that produces polymer materials and to improve their environmental performance. The results obtained show that the whole productive process has a low environmental impact factor. Nevertheless the applied methodology puts in evidence some processes that generate local pollution in specific areas of the factory and which could be dangerous for the workers' health. PMID- 15531387 TI - Removal of Fe(II) from the wastewater of a galvanized pipe manufacturing industry by adsorption onto bentonite clay. AB - Bentonite clay has been used for the adsorption of Fe(II) from aqueous solutions over a concentration range of 80-200 mg/l, shaking time of 1-60 min, adsorbent dosage from 0.02 to 2 g and pH of 3. The process of uptake follows both the Langmuir and Freundlich isotherm models and also the first-order kinetics. The maximum removal (>98%) was observed at pH of 3 with initial concentration of 100 mg/l and 0.5 g of bentonite. The efficiency of Fe(II) removal was also tested using wastewater from a galvanized pipe manufacturing industry. More than 90% of Fe(II) can be effectively removed from the wastewater by using 2.0 g of the bentonite. The effect of cations (i.e. zinc, manganese, lead, cadmium, nickel, cobalt, chromium and copper) on the removal of Fe(II) was studied in the concentration range of 10-500 mg/l. All the added cations reduced the adsorption of Fe(II) at high concentrations except Zn. Column studies have also been carried out using a certain concentration of wastewater. More than 99% recovery has been achieved by using 5 g of the bentonite with 3M nitric acid solution. PMID- 15531388 TI - Aggregate eco-efficiency indices for New Zealand--a principal components analysis. AB - Eco-efficiency has emerged as a management response to waste issues associated with current production processes. Despite the popularity of the term in both business and government circles, limited attention has been paid to measuring and reporting eco-efficiency to government policy makers. Aggregate measures of eco efficiency are needed, to complement existing measures and to help highlight important patterns in eco-efficiency data. This paper aims to develop aggregate measures of eco-efficiency for use by policy makers. Specifically, this paper provides a unique analysis by applying principal components analysis (PCA) to eco efficiency indicators in New Zealand. The study reveals that New Zealand's overall eco-efficiency improved for two out of the five aggregate measures over the period 1994/1995-1997/1998. The worsening of the other aggregate measures reflects, among other things, the relatively poor performance of the primary production and related processing sectors. These results show PCA is an effective approach for aggregating eco-efficiency indicators and assisting decision makers by reducing redundancy in an eco-efficiency indicators matrix. PMID- 15531389 TI - Roofing as a source of nonpoint water pollution. AB - Sixteen wooden structures with two roofs each were installed to study runoff quality for four commonly used roofing materials (wood shingle, composition shingle, painted aluminum, and galvanized iron) at Nacogdoches, Texas. Each roof, either facing NW or SE, was 1.22 m wide x 3.66 m long with a 25.8% roof slope. Thus, there were 32 alternatively arranged roofs, consisting of four roof types x two aspects x four replicates, in the study. Runoff from the roofs was collected through galvanized gutters, downspouts, and splitters. The roof runoff was compared to rainwater collected by a wet/dry acid rain collector for the concentrations of eight water quality variables, i.e. Cu(2+), Mn(2+), Pb(2+), Zn(2+), Mg(2+), Al(3+), EC and pH. Based on 31 storms collected between October 1997 and December 1998, the results showed: (1) concentrations of pH, Cu, and Zn in rainwater already exceed the EPA freshwater quality standards even without pollutant inputs from roofs, (2) Zn and Cu, the two most serious pollutants in roof runoff, exceeded the EPA national freshwater water quality standards in virtually 100% and more than 60% of the samples, respectively, (3) pH, EC, and Zn were the only three variables significantly affected by roofing materials, (4) differences in Zn concentrations were significant among all roof types and between all roof runoff and rainwater samples, (5) although there were no differences in Cu concentrations among all roof types and between roof runoff and rainwater, all means and medians of runoff and rainwater exceeded the national water quality standards, (6) water quality from wood shingles was the worst among the roof types studied, and (7) although SE is the most frequent and NW the least frequent direction for incoming storms, only EC, Mg, Mn, and Zn in wood shingle runoff from the SE were significantly higher than those from the NW; the two aspects affected no other elements in runoff from the other three roof types. Also, Zn concentrations from new wood-shingle roofs were significantly higher than those from aged roofs of a previous study. The study demonstrated that roofs could be a serious source of nonpoint water pollution. Since Zn is the most serious water pollutant and wood shingle is the worst of the four roof types, using less compounds and materials associated with Zn along with good care and maintenance of roofs are critical in reducing Zn pollution in roof runoff. PMID- 15531390 TI - Scope insensitivity in contingent valuation of complex environmental amenities. AB - It has been argued that respondents in contingent valuation (CV) surveys, asked to value complex environmental amenities, will state willingness to pay (WTP) independently of the scope of the project. Such insensitivity to scope would be at odds with rational choice, and could therefore imply that CV is not a theoretically valid method for biodiversity valuation. The scope test in the present CV study was applied to endangered species preservation. Respondents were split in four sub-samples facing different scopes of endangered species preservation. The design allowed for both external and internal scope tests. Furthermore, the tests were split according to elicitation format. Of four external tests of insensitivity to scope, one was rejected, two gave mixed results, depending on either the type of test or elicitation format, and for the last one the null hypothesis could not be rejected. Of five internal tests, insensitivity to scope was rejected in three cases, one test gave mixed results, and one could not be rejected. Survey design features of the CV study, especially an unfamiliar sub-group of endangered species, could explain the apparent insensitivity to scope observed. PMID- 15531391 TI - Gaseous emissions from lignosulfonates in dust abatement applications. AB - Concerns over possible gaseous emissions from lignosulfonate dust suppressant agents used on dirt roads prompted an investigation into the possible release of sulfur dioxide and organic vapors. Emissions from eight commercial lignosulfonates products applied to three different soil types were measured in environmental chambers. It was determined that except under extreme soil pH conditions (<1), the release of sulfur dioxide from the application of lignosulfonates to roads is essentially non existent. The odor associated with the application was found to be due to trace amounts of organic compounds mainly of the furfural variety. Levels of odor compounds in these products was significantly lower than the previously reported values for spent sulfite liquors because the majority were removed during processing operations prior to sale. PMID- 15531392 TI - Cleaner production in the ammonia-soda industry: an ecological and economic study. AB - Five methods to reduce the negative influence of soda ash factories on the natural environment are presented: 1. obtaining calcium-magnesium phosphates by treating the suspension from raw brine purification with orthophosphoric acid (H(3)PO(4)), 2. production of precipitated chalk from soda processing waste, 3. production of gypsum and semi-brine, 4. desulphurisation of fume gases from the factory power plant, 5. utilization of distiller waste. The tests, accomplished on a laboratory scale, showed the high efficiency of these methods. Economic analysis has proved that only four out of the five presented processes can have a positive financial effect on soda ash factories, as well as being well justified economically. The value of two of the innovations presented is confirmed by their implementation in factories. PMID- 15531393 TI - Sustainability indicator system and policy processes in Malaysia: a framework for utilisation and learning. AB - Formulation of effective sustainability indicators for national assessment demands a comprehensive understanding of the utilisation, diffusion and dissemination of information in policy processes. To illustrate the dynamic of sustainability assessment within the context of policy processes, this paper uses a case study of national sustainability indicators development in Malaysia. Subsequently, this paper ascribes the limited achievement of national sustainability assessment in Malaysia to four types of constraints: meta-policy issues; technical capacities; communication concerns; and the inherent knowledge gaps within the indicator developer community vis-a-vis their theoretical limitations. It is proposed that such constraints will be encountered in many countries. Drawing from the literature on public policy, this paper outlines a framework for investigating indicator behaviour within policy processes based on well-established concepts such as knowledge utilisation and policy learning. I conclude this paper by elaborating on the corresponding future challenges that must be addressed before effective integration of sustainability indicators within policy systems can occur. PMID- 15531394 TI - Partial breast irradiation: revolution or evolution? AB - INTRODUCTION: It is well established that there is no significant difference in terms of overall survival between mastectomy and radio-surgical conservative treatment (RSCT). Ten to fifteen years have been needed to change from the "standard" to the "new" breast cancer treatment. At the end of the 1990s, a few authors published preliminary results of phase II trials using partial breast irradiation (PBI) as sole post-operative treatment based on interstitial low or high dose rate brachytherapy techniques. Currently, phase III randomized trials comparing whole breast irradiation versus PBI are ongoing in the USA and in EU. MATERIALS AND METHODS: The authors reviewed the literature regarding issues in the irradiation of breast cancer and the potential role of PBI to prevent local recurrence in the ipsilateral breast. Furthermore, potential advantages of PBI were discussed as well as the limits of such a procedure. Finally, we tried to determine the characteristics of a subgroup of patients who would receive benefit from this radiation technique. RESULTS: Phase II and III trials have been analyzed concerning feasibility, efficacy and toxicity. PBI may be delivered through low or high dose rate brachytherapy or intraoperative/external beam radiation therapy. PBI satisfies the control quality criteria. The majority of the teams provide PBI recurrence rates lower than 5% (0-4.4%) with a median follow-up varying between 8 and 72 months, and associated with cosmetic results comparable to those achieved with conventional external beam. CONCLUSION: RSCT does not replace mastectomy; mastectomy and RSCT are used together to treat different subgroups of patients. PBI furnished encouraging preliminary results in terms of local control and cosmetic results. However, caution in interpreting the results is required along with longer follow-up. PBI should not replace CT, but could be considered as a new therapeutic strategy for breast cancer dedicated to a very well defined subgroup of patients with a low risk of local recurrence. PMID- 15531395 TI - Effects of statins and farnesyltransferase inhibitors on the development and progression of cancer. AB - Statins (3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors - HMG-CoA reductase inhibitors) have been approved for the treatment of lipid disorders. Recently, in vivo studies with experimental animals and in vitro studies indicated a possible role for statins in the treatment of malignancies. Inhibition of the enzyme HMG-CoA reductase results in decreased farnesylation and geranylgeranylation of several proteins essential for cellular proliferation and survival. Inhibition of Ras farnesylation was originally thought to be the mechanism that mediates statin-induced effects in cancer. Consequently, specific inhibitors of the enzyme farnesyltransferase (FTIs) were developed. Currently, the mechanisms that mediate statin- and FTI-induced antitumour effects are questioned. It remains unclear which proteins and signal transduction cascades are involved. This review focuses on the effects and possible therapeutic application of statins and FTIs. Antitumour properties such as induction of growth arrest and apoptosis, inhibition of metastasis and inhibition of angiogenesis are discussed. Furthermore, the mechanisms of statin- and farnesyltransferase inhibitor-induced effects and the involvement of a number of cellular components (such as farnesylated and geranylgeranylated proteins, the mitogen-activated protein kinase signalling pathway, the phosphoinositide 3' kinase signalling pathway, and cell cycle regulatory proteins) are reviewed. In addition, clinical and epidemiological data with respect to statins and farnesyltransferase inhibitors are summarised. We propose that inhibitors of the mevalonate pathway are particularly effective when administered in combination with other drugs. Therefore, the mechanisms and effects of combined therapy of statins or farnesyltransferase inhibitors with chemotherapeutics, biphosphonates, non-steroidal anti-inflammatory drugs, specific inhibitors of geranylgeranyltransferase and inhibitors of tyrosine kinase activity are discussed. PMID- 15531396 TI - Evaluating the role of dexrazoxane as a cardioprotectant in cancer patients receiving anthracyclines. AB - Anthracyclines remain an important group of chemotherapeutic agents, despite their inherent cardiotoxicity. This cardiotoxicity may be even more of a concern in the future, as combination therapies of anthracyclines with newer agents become routine. Such combinations may be highly effective, but cardiotoxicity may also be increased. Dexrazoxane reduces the incidence of cardiotoxicity, as demonstrated in numerous clinical trials in both adults and children. Evidence from the literature suggests no effect of dexrazoxane on the antitumour efficacy of anthracyclines, and there is no adverse effect on survival. Dexrazoxane is therefore a valuable tool for oncologists using anthracycline-based regimens. PMID- 15531397 TI - Gefitinib does not improve survival when combined with gemcitabine and cisplatin in people with advanced non-small-cell lung cancer. PMID- 15531398 TI - Gefitinib does not improve survival when combined with paclitaxel and carboplatin chemotherapy in people with advanced non-small-cell lung cancer. PMID- 15531399 TI - Adjuvant chemotherapy with uracil-tegafur prolongs overall survival in people with primary adenocarcinoma of the lung. PMID- 15531400 TI - High-quality evidence in oncology from 20th April to 11th May 2004: a summary. PMID- 15531401 TI - The MATRICS initiative: developing a consensus cognitive battery for clinical trials. PMID- 15531402 TI - Measurement and Treatment Research to Improve Cognition in Schizophrenia: NIMH MATRICS initiative to support the development of agents for improving cognition in schizophrenia. AB - The impairments in social and vocational outcome that are common in schizophrenia are strongly related to the severity of impaired neurocognition. This observation led to the initiation of The NIMH's Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) initiative, which supports the development of pharmacological agents to improve cognition in schizophrenia. MATRICS addresses barriers to drug development through a number of activities, including the development of a consensus battery for measuring cognition in schizophrenia; the development of a consensus regarding the most promising molecular targets that should be the focus of drug development; the use of a joint meeting with representatives from the industry, academia, NIMH, and the U.S. Food and Drug Administration (FDA) to clarify guidelines for the design of clinical trials for cognition enhancing agents; and finally, to assist NIMH in developing its research agenda in this area. PMID- 15531403 TI - NIMH-MATRICS survey on assessment of neurocognition in schizophrenia. AB - The NIMH-Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) initiative requires, among other things, the establishment of a reliable, valid, and consensus-derived method of assessing cognition. The derived battery will provide a standardized way to assess the effects of cognition enhancing agents across clinical trials. To this end, the first of six consensus oriented conferences was held April 2003. The goals were twofold: (a) To select which cognitive constructs to measure in a consensus battery, and (b) to select which criteria to use in evaluating tests for inclusion in the battery. Based on consultation with experts on the RAND Panel Method, 74 experts were invited to participate in a pre-meeting survey to provide information relevant to decisions on the cognitive battery. The survey included sections on reliability, validity, test administration, norms and interpretation of tests, cognitive domains and their integration, battery duration, and overall importance of test qualities. For selection of cognitive targets, the results showed that experts ranked executive functions, attention/vigilance, memory processes, and problem-solving ability highest. For test qualities, the experts ranked test-retest reliability, good coverage of key individual cognitive constructs, and comparable alternate forms highest. This article presents the results of the pre-conference survey that was the first step in the RAND process towards development of the NIMH MATRICS consensus battery to assess cognition in schizophrenia. PMID- 15531404 TI - Cognitive deficits as treatment targets in schizophrenia. AB - Cognitive impairment has emerged as an important new target in schizophrenia therapeutics in light of evidence that cognitive deficits are critically related to the functional of disability that is characteristic of the illness. Evidence is briefly reviewed supporting the idea that the cognitive impairment in schizophrenia is an attractive target for therapeutic intervention including: (1) there is a characteristic pattern of cognitive deficits that occur with very high frequency; (2) the deficits are relatively stable over time; and (3) cognitive deficits are relatively independent of the symptomatic manifestations of the illness. Thus, cognitive impairment appears to be a well-defined, reliable and distinct dimension of the illness. PMID- 15531405 TI - Identification of separable cognitive factors in schizophrenia. AB - One of the primary goals in the NIMH initiative to encourage development of new interventions for cognitive deficits in schizophrenia, Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS), has been to develop a reliable and valid consensus cognitive battery for use in clinical trials. Absence of such a battery has hampered standardized evaluation of new treatments and, in the case of pharmacological agents, has been an obstacle to FDA approval of medications targeting cognitive deficits in schizophrenia. A fundamental step in developing such a battery was to identify the major separable cognitive impairments in schizophrenia. As part of this effort, we evaluated the empirical evidence for cognitive performance dimensions in schizophrenia, emphasizing factor analytic studies. We concluded that seven separable cognitive factors were replicable across studies and represent fundamental dimensions of cognitive deficit in schizophrenia: Speed of Processing, Attention/Vigilance, Working Memory, Verbal Learning and Memory, Visual Learning and Memory, Reasoning and Problem Solving, and Verbal Comprehension. An eighth domain, Social Cognition, was added due to recent increased interest in this area and other evidence of its relevance for clinical trials aiming to evaluate the impact of potential cognitive enhancers on cognitive performance and functional outcome. Verbal Comprehension was not considered appropriate for a cognitive battery intended to be sensitive to cognitive change, due to its resistance to change. The remaining seven domains were recommended for inclusion in the MATRICS-NIMH consensus cognitive battery and will serve as the basic structure for that battery. These separable cognitive dimensions also have broader relevance to future research aimed at understanding the nature and structure of core cognitive deficits in schizophrenia. PMID- 15531406 TI - Longitudinal studies of cognition and functional outcome in schizophrenia: implications for MATRICS. AB - It is generally accepted that cognitive deficits in schizophrenia are related to functional outcome. However, support for longitudinal relationships between cognition and functional outcome has not been as well documented. The current paper presents a review of 18 recently published longitudinal studies (minimum 6 month follow up) of the relationships between cognition and community outcome in schizophrenia. Results from these studies reveal considerable support for longitudinal associations between cognition and community outcome in schizophrenia. These studies demonstrate that cognitive assessment predict later functional outcome and provide a rationale for psychopharmacological interventions for cognitive deficits in schizophrenia. Although the relationships between cognition and community outcome are well-supported, it is clear that community functioning is also affected by a host of factors apart from cognition that are usually not considered in clinical trial studies (e.g., psychosocial rehabilitation and educational/vocational opportunities). In the second part of the paper, we consider intervening steps between cognitive performance measures and community outcome. These steps are apt to have important implications for clinical trials of cognition-enhancing agents in schizophrenia. PMID- 15531408 TI - Clinical constructs and therapeutic discovery. AB - Schizophrenia, as a single disease entity, has been the major paradigm during the past 100 years. Classification has increasingly emphasized diagnostically discriminating psychotic symptoms. This has made reality distortion and thought disorder the defining qualities of the disorder. One result is 50 years of developing dopamine antagonist antipsychotic drugs while neglecting novel therapeutic pathways related to nonpsychotic pathology. The domains of psychopathology provides an alternative paradigm. In this paradigm, the relative independence of reality distortion, disorganization, negative pathology, and impairments in cognition is stressed. Application of this paradigm has revealed that impaired cognition and negative symptoms are robustly associated with poor functional outcomes. Antipsychotic drugs fail to address these pathologies. These unmet treatment needs are clinical targets for drug discovery involving novel therapeutic pathways. PMID- 15531407 TI - Direct assessment of functional abilities: relevance to persons with schizophrenia. AB - OBJECTIVE: To review functional outcome assessment strategies specifically focusing on performance-based measures used in patients with schizophrenia. METHOD: In order to identify relevant performance-based measures of functioning we conducted a literature search using key words: functioning, disability, performance, situation, assessment, skills, scale, measurement, instrument, psychosis, and schizophrenia. RESULTS: A description of each of eight measures was provided including assessment protocol, domains of functioning assessed, scoring, basic psychometric properties, appropriateness for clinical trials and likely sensitivity to pharmacologic interventions. Methods for assessment of functioning in patients with schizophrenia are reviewed. CONCLUSIONS: The measurement of functional capacity, through performance-based activities, provides a critical benchmark by which to evaluate the efficacy and effectiveness of medications in clinical trials to improve both cognition and capacity for everyday functioning in seriously mentally ill persons with schizophrenia. In order to comprehensively evaluate functional outcome, a combination of performance-based and other assessment modalities may be required. PMID- 15531409 TI - Program to improve cognitive functioning in patients with schizophrenia: reflections. AB - Cognitive dysfunction in patients who suffer from schizophrenia is a major clinical problem. For such patients, the neurobiological cause(s) for the cognition deficits are not clear. Treatments with high doses of compounds that work at the coagonist site of the NMDA receptor have demonstrated that some degree of improvement can be elicited in patients. Thus, the NIMH effort to delineate the clinical responses that would allow new drugs to be registered for this use is very timely. Significant credit is due Dr. Steven Hyman and his colleagues for stimulating this bold initiative. Many problems remain to be solved to accomplish the goal. This article articulates a few such problems. PMID- 15531410 TI - Environment and health: capacity building for the future. PMID- 15531411 TI - Update of residential tetrachloroethylene exposure and decreases in visual contrast sensitivity. PMID- 15531413 TI - Are maternal thyroid autoantibodies generated by PCBs the missing link to impaired development of the brain? PMID- 15531415 TI - Agricultural task not predictive of children's exposure to OP pesticides. PMID- 15531416 TI - Pesticides and organic agriculture. PMID- 15531417 TI - More recent studies on fragrances. PMID- 15531418 TI - Olden's contributions. PMID- 15531421 TI - National Toxicology Program: landmarks and the road ahead. PMID- 15531422 TI - Environmental cardiology: getting to the heart of the matter. PMID- 15531423 TI - Coal: poised for a comeback? PMID- 15531424 TI - Underwater logging: Submarine rediscovers lost wood. PMID- 15531427 TI - Listing occupational carcinogens. AB - The occupational environment has been a most fruitful one for investigating the etiology of human cancer. Many recognized human carcinogens are occupational carcinogens. There is a large volume of epidemiologic and experimental data concerning cancer risks in different work environments. It is important to synthesize this information for both scientific and public health purposes. Various organizations and individuals have published lists of occupational carcinogens. However, such lists have been limited by unclear criteria for which recognized carcinogens should be considered occupational carcinogens, and by inconsistent and incomplete information on the occupations and industries in which the carcinogenic substances may be found and on their target sites of cancer. Based largely on the evaluations published by the International Agency for Research on Cancer, and augmented with additional information, the present article represents an attempt to summarize, in tabular form, current knowledge on occupational carcinogens, the occupations and industries in which they are found, and their target organs. We have considered 28 agents as definite occupational carcinogens, 27 agents as probable occupational carcinogens, and 113 agents as possible occupational carcinogens. These tables should be useful for regulatory or preventive purposes and for scientific purposes in research priority setting and in understanding carcinogenesis. PMID- 15531428 TI - Organochlorine exposure and colorectal cancer risk. AB - Organochlorine compounds have been linked to increased risk of several cancers. Despite reductions in their use and fugitive release, they remain one of the most important groups of persistent pollutants to which humans are exposed, primarily through dietary intake. We designed a case-control study to assess the risk of colorectal cancer with exposure to these chemicals, and their potential interactions with genetic alterations in the tumors. A subsample of cases (n = 132) and hospital controls (n = 76) was selected from a larger case-control study in Barcelona, Catalonia, Spain. We measured concentrations in serum of several organochlorines by gas chromatography. We assessed point mutations in K-ras and p53 genes in tissue samples by polymerase chain reaction/single-strand conformation polymorphism and assessed expression of p53 protein by immunohistochemical methods. An elevated risk of colorectal cancer was associated with higher serum concentrations of mono-ortho polychlorinated biphenyl (PCB) congeners 28 and 118. The odds ratio for these mono-ortho PCBs for middle and higher tertile were, respectively, 1.82 [95% confidence interval (CI), 0.90-3.70] and 2.94 (95% CI, 1.39-6.20). Alpha-hexachlorocyclohexane, hexachlorobenzene, and p,p'-DDE (4,4'-dichlorodiphenyltrichloroethene) showed nonsignificant increases in risk. Risk associated with mono-ortho PCBs was slightly higher for tumors with mutations in the p53 gene but was not modified by mutations in K-ras. Mono-ortho PCBs were further associated with transversion-type mutations in both genes. These results generate the hypothesis that exposure to mono-ortho PCBs contributes to human colorectal cancer development. The trend and magnitude of the association, as well as the observation of a molecular fingerprint in tumors, raise the possibility that this finding may be causal. PMID- 15531429 TI - Androgenic and estrogenic response of green mussel extracts from Singapore's coastal environment using a human cell-based bioassay. AB - In the last decade, evidence of endocrine disruption in biota exposed to environmental pollutants has raised serious concern. Human cell-based bioassays have been developed to evaluate induced androgenic and estrogenic activities of chemical compounds. However, bioassays have been sparsely applied to environmental samples. In this study we present data on sex hormone activities in the green mussel, Perna viridis, in Singapore's coastal waters. P.viridis is a common bioindicator of marine contamination, and this study is a follow-up to an earlier investigation that reported the presence of sex hormone activities in seawater samples from Singapore's coastal environment. Specimens were collected from eight locations around the Singapore coastline and analyzed for persistent organic pollutants (POPs) and heavy metals. Tissue extracts were then screened for activities on androgen receptors (ARs) and estrogen receptors (ER-alpha and ER-beta) using a reporter gene bioassay based on a HeLa human cell line. Mussel extracts alone did not exhibit AR activity, but in the presence of the reference androgenic hormone dihydrotestosterone (DHT), activities were up to 340% higher than those observed for DHT alone. Peak activities were observed in locations adjacent to industrial and shipping activities. Estrogenic activities of the mussel extract both alone and in the presence of reference hormone were positive. Correlations were statistically investigated between sex hormone activities, levels of pollutants in the mussel tissues, and various biological parameters (specimen size, sex ratio, lipid and moisture content). Significant correlations exist between AR activities, in the presence of DHT, and total concentration of POPs (r= 0.725, p < 0.05). PMID- 15531430 TI - Cumulative dietary energy intake determines the onset of puberty in female rats. AB - Laboratory animal diets for studies to determine the endocrine-disrupting potential of chemicals are under scrutiny because they can affect both assay control values and assay sensitivity. Although phytoestrogen content is important, we have previously shown that a phytoestrogen-rich diet and a phytoestrogen-free diet were equally uterotrophic to rats and advanced vaginal opening (VO) when compared with the standard diet RM1. Abolition of the effects by the gonadotrophin-releasing hormone antagonist Antarelix indicated that these effects were mediated through the hypothalamus-pituitary-reproductive organ axis. In the present study, we investigated the relationship between cumulative energy intake and sexual maturation in female rats. Infant formula (IF) at different concentrations and synthetic diets, with a wide range of metabolizable energy (ME) values, were used to modulate energy intake. Increasing energy intake was associated with an increase in uterine weight (absolute and adjusted for body weight) for both IF and the synthetic diets. In both cases, the increased uterine weight was directly proportional to energy intake. Body weight was unaffected by IF consumption but, in the case of the diets, was increased proportionally with energy consumption. Antarelix abolished the uterine weight increases with both formula and the diets, whereas body weight was unaffected. The mean day of VO was also advanced by high-ME diets and IF, whereas body weight at VO was unaffected. VO occurred at an energy intake of approximately 2,300 kJ/rat determined by measuring total food intake from weaning to VO, indicating that this cumulative energy intake was the trigger for puberty. ME is therefore a critical factor in the choice of diets for endocrine disruption studies. PMID- 15531431 TI - Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways. AB - Xenoestrogens can mimic or antagonize the activity of physiological estrogens, and the suggested mechanism of xenoestrogen action involves binding to estrogen receptors (ERs). However, the failure of various in vitro or in vivo assays to show strong genomic activity of xenoestrogens compared with estradiol (E2) makes it difficult to explain their ability to cause abnormalities in animal (and perhaps human) reproductive functions via this pathway of steroid action. E2 has also been shown to initiate rapid intracellular signaling, such as changes in levels of intracellular calcium, cAMP, and nitric oxide, and activations of a variety of kinases, via action at the membrane. In this study, we demonstrate that several xenoestrogens can rapidly activate extracellular-regulated kinases (ERKs) in the pituitary tumor cell line GH3/B6/F10, which expresses high levels of the membrane receptor for ER-alpha (mER). We tested a phytoestrogen (coumestrol), organochlorine pesticides or their metabolites (endosulfan, dieldrin, and DDE), and detergent by-products of plastics manufacturing (p nonylphenol and bisphenol A). These xenoestrogens (except bisphenolA) produced rapid (3-30 min after application), concentration (10(-14)-10(-8) M)-dependent ERK-1/2 phosphorylation but with distinctly different activation patterns. To identify signaling pathways involved in ERK activation, we used specific inhibitors of ERs, epidermal growth factor receptors, Ca2+ signaling, Src and phosphoinositide-3 kinases, and a membrane structure disruption agent. Multiple inhibitors blocked ERK activation, suggesting simultaneous use of multiple pathways and complex signaling web interactions. However, inhibitors differentially affected each xenoestrogen response examined. These actions may help to explain the distinct abilities of xenoestrogens to disrupt reproductive functions at low concentrations. PMID- 15531432 TI - Effects of 1,3-butadiene, isoprene, and their photochemical degradation products on human lung cells. AB - Because of potential exposure both in the workplace and from ambient air, the known carcinogen 1,3-butadiene (BD) is considered a priority hazardous air pollutant. BD and its 2-methyl analog, isoprene (ISO), are chemically similar but have very different toxicities, with ISO showing no significant carcinogenesis. Once released into the atmosphere, reactions with species induced by sunlight and nitrogen oxides convert BD and ISO into several photochemical reaction products. In this study, we determined the relative toxicity and inflammatory gene expression induced by exposure of A549 cells to BD, ISO, and their photochemical degradation products in the presence of nitric oxide. Gas chromatography and mass spectrometry analyses indicate the initial and major photochemical products produced during these experiments for BD are acrolein, acetaldehyde, and formaldehyde, and products for ISO are methacrolein, methyl vinyl ketone, and formaldehyde; both formed < 200 ppb of ozone. After exposure the cells were examined for cytotoxicity and interleukin-8 (IL-8) gene expression, as a marker for inflammation. These results indicate that although BD and ISO alone caused similar cytotoxicity and IL-8 responses compared with the air control, their photochemical products significantly enhanced cytotoxicity and IL-8 gene expression. This suggests that once ISO and BD are released into the environment, reactions occurring in the atmosphere transform these hydrocarbons into products that induce potentially greater adverse health effects than the emitted hydrocarbons by themselves. In addition, the data suggest that based on the carbon concentration or per carbon basis, biogenic ISO transforms into products with proinflammatory potential similar to that of BD products. PMID- 15531433 TI - Lead sources in human diet in Greenland. AB - Although blood lead levels have declined in Greenland, they are still elevated despite the fact that lead levels in the Greenland environment are very low. Fragments of lead shot in game birds have been suggested as an important source of dietary exposure, and meals of sea birds, particularly eider, contain high concentrations of lead. In a cross-sectional population survey in Greenland in 1993-1994, blood lead adjusted for age and sex was found to be associated with the reported consumption of sea birds. Participants reporting less than weekly intake of sea birds had blood lead concentrations of approximately 75 microg/L, whereas those who reported eating sea birds several times a week had concentrations of approximately 110 microg/L, and those who reported daily intake had concentrations of 170 microg/L (p = 0.01). Blood lead was not associated with dietary exposure to other local or imported food items. PMID- 15531434 TI - Blood lead changes during pregnancy and postpartum with calcium supplementation. AB - Pregnancy and lactation are times of physiologic stress during which bone turnover is accelerated. Previous studies have demonstrated that there is increased mobilization of lead from the maternal skeleton at this time and that calcium supplementation may have a protective effect. Ten immigrants to Australia were provided with either calcium carbonate or a complex calcium supplement (approximately 1 g/day) during pregnancy and for 6 months postpartum. Two immigrant subjects who did not conceive acted as controls. Sampling involved monthly venous blood samples throughout pregnancy and every 2 months postpartum, and quarterly environmental samples and 6-day duplicate diets. The geometric mean blood lead at the time of first sampling was 2.4 microg/dL (range, 1.4-6.5). Increases in blood lead during the third trimester, corrected for hematocrit, compared with the minimum value observed, varied from 10 to 50%, with a geometric mean of 25%. The increases generally occurred at 6-8 months gestation, in contrast with that found for a previous cohort, characterized by very low calcium intakes, where the increases occurred at 3-6 months. Large increases in blood lead concentration were found during the postpartum period compared with those during pregnancy; blood lead concentrations increased by between 30 and 95% (geometric mean 65%; n = 8) from the minimum value observed during late pregnancy. From late pregnancy through postpartum, there were significant increases in the lead isotopic ratios from the minimum value observed during late pregnancy for 3 of 8 subjects (p < 0.01). The observed changes are considered to reflect increases in mobilization of lead from the skeleton despite calcium supplementation. The identical isotopic ratios in maternal and cord blood provide further confirmation of placental transfer of lead. The extra flux released from bone during late pregnancy and postpartum varies from 50 to 380 microg lead (geometric mean, 145 microg lead) compared with 330 microg lead in the previous cohort. For subjects replete in calcium, the delay in increase in blood lead and halving of the extra flux released from bone during late pregnancy and postpartum may provide less lead exposure to the developing fetus and newly born infant. Nevertheless, as shown in several other studies on calcium relationships with bone turnover, calcium supplementation appears to provide limited benefit for lead toxicity during lactation. PMID- 15531435 TI - Production of androgens by microbial transformation of progesterone in vitro: a model for androgen production in rivers. AB - We have previously documented the presence of progesterone and androstenedione in the water column and bottom sediments of the Fenholloway River, Taylor County, Florida. This river receives paper mill effluent and contains masculinized female mosquitofish. We hypothesized that plant sterols (e.g., ss-sitosterol) derived from the pulping of pine trees are transformed by bacteria into progesterone and subsequently into 17alpha-hydroxyprogesterone, androstenedione, and other androgens. In this study, we demonstrate that these same androgens can be produced in vitro from the bacterium Mycobacterium smegmatis. In a second part to this study, we reextracted and reanalyzed the sediment from the Fenholloway River and verified the presence of androstadienedione, a delta1 steroid with androgen activity. PMID- 15531436 TI - Evidence for concurrent effects of exposure to environmental cadmium and lead on hepatic CYP2A6 phenotype and renal function biomarkers in nonsmokers. AB - We examined the interrelationships between phenotype of hepatic cytochrome P450 2A6 (CYP2A6), nephropathy, and exposure to cadmium and lead in a group of 118 healthy Thai men and women who had never smoked. Their urinary Cd excretion ranged from 0.05 to 2.36 microg/g creatinine, whereas their urinary Pb excretion ranged from 0.1 to 12 microg/g creatinine. Average age and Cd burden of women and men did not differ. Women, however, on average showed a 46% higher urinary Pb excretion (p < 0.001) and lower zinc status, suggested by lower average serum Zn and urinary Zn excretion compared with those in men. Cd-linked nephropathy was detected in both men and women. However, Pb-linked nephropathy was seen only in women, possibly because of higher Pb burden coupled with lower protective factors, notably of Zn (p < 0.001), in women compared with men. In men, Pb burden showed a negative association with CYP2A6 activity (adjusted beta = -0.29, p = 0.003), whereas Cd burden showed a positive association with CYP2A6 activity (adjusted beta = 0.38, p = 0.001), suggesting opposing effects of Cd and Pb on hepatic CYP2A6 phenotype. The weaker correlation between Cd burden CYP2A6 activity in women despite similarity in Cd burden between men and women is consistent with opposing effects of Pb and Cd on hepatic CYP2A6 phenotypic expression. A positive correlation between Cd-linked nephropathy (urinary N acetyl-beta-D-glucosaminidase excretion) and CYP2A6 activity in men (r = 0.39, p = 0.002) and women (r = 0.37, p = 0.001) suggests that Cd induction of hepatic CYP2A6 expression and Cd-linked nephropathy occurred simultaneously. PMID- 15531437 TI - Impact of the phytoestrogen content of laboratory animal feed on the gene expression profile of the reproductive system in the immature female rat. AB - The effect of the dietary background of phytoestrogens on the outcome of rodent bioassays used to identify and assess the reproductive hazard of endocrine disrupting chemicals is controversial. Phytoestrogens, including genistein, daidzein, and coumestrol, are fairly abundant in soybeans and alfalfa, common ingredients of laboratory animal diets. These compounds are weak agonists for the estrogen receptor (ER) and, when administered at sufficient doses, elicit an estrogenic response in vivo. In this study, we assessed the potential estrogenic effects of dietary phytoestrogens at the gene expression level, together with traditional biologic end points, using estrogen-responsive tissues of the immature female rat. We compared the gene expression profile of the uterus and ovaries, as a pool, obtained using a uterotrophic assay protocol, from intact prepubertal rats fed a casein-based diet (free from soy and alfalfa) or a regular rodent diet (Purina 5001) containing soy and alfalfa. Estrogenic potency of the phytoestrogen-containing diet was determined by analyzing uterine wet weight gain, luminal epithelial cell height, and gene expression profile in the uterus and ovaries. These were compared with the same parameters evaluated in animals exposed to a low dose of a potent ER agonist [0.1 microg/kg/day 17alpha-ethynyl estradiol (EE) for 4 days]. Exposure to dietary phytoestrogens or to a low dose of EE did not advance vaginal opening, increase uterine wet weight, or increase luminal epithelial cell height in animals fed either diet. Although there are genes whose expression differs in animals fed the soy/alfalfa-based diet versus the casein diet, those genes are not associated with estrogenic stimulation. The expression of genes well known to be estrogen regulated, such as progesterone receptor, intestinal calcium-binding protein, and complement component 3, is not affected by consumption of the soy/alfalfa-based diet when assessed by microarray or quantitative reverse transcriptase-polymerase chain reaction analysis. Our results indicate that although diet composition has an impact on gene expression in uterus and ovaries, it does not contribute to the effects of an ER agonist. PMID- 15531438 TI - Relationship between composition and toxicity of motor vehicle emission samples. AB - In this study we investigated the statistical relationship between particle and semivolatile organic chemical constituents in gasoline and diesel vehicle exhaust samples, and toxicity as measured by inflammation and tissue damage in rat lungs and mutagenicity in bacteria. Exhaust samples were collected from "normal" and "high-emitting" gasoline and diesel light-duty vehicles. We employed a combination of principal component analysis (PCA) and partial least-squares regression (PLS; also known as projection to latent structures) to evaluate the relationships between chemical composition of vehicle exhaust and toxicity. The PLS analysis revealed the chemical constituents covarying most strongly with toxicity and produced models predicting the relative toxicity of the samples with good accuracy. The specific nitro-polycyclic aromatic hydrocarbons important for mutagenicity were the same chemicals that have been implicated by decades of bioassay-directed fractionation. These chemicals were not related to lung toxicity, which was associated with organic carbon and select organic compounds that are present in lubricating oil. The results demonstrate the utility of the PCA/PLS approach for evaluating composition-response relationships in complex mixture exposures and also provide a starting point for confirming causality and determining the mechanisms of the lung effects. PMID- 15531439 TI - Lung cancer in railroad workers exposed to diesel exhaust. AB - Diesel exhaust has been suspected to be a lung carcinogen. The assessment of this lung cancer risk has been limited by lack of studies of exposed workers followed for many years. In this study, we assessed lung cancer mortality in 54,973 U.S. railroad workers between 1959 and 1996 (38 years). By 1959, the U.S. railroad industry had largely converted from coal-fired to diesel-powered locomotives. We obtained work histories from the U.S. Railroad Retirement Board, and ascertained mortality using Railroad Retirement Board, Social Security, and Health Care Financing Administration records. Cause of death was obtained from the National Death Index and death certificates. There were 43,593 total deaths including 4,351 lung cancer deaths. Adjusting for a healthy worker survivor effect and age, railroad workers in jobs associated with operating trains had a relative risk of lung cancer mortality of 1.40 (95% confidence interval, 1.30-1.51). Lung cancer mortality did not increase with increasing years of work in these jobs. Lung cancer mortality was elevated in jobs associated with work on trains powered by diesel locomotives. Although a contribution from exposure to coal combustion products before 1959 cannot be excluded, these results suggest that exposure to diesel exhaust contributed to lung cancer mortality in this cohort. Key words: diesel exhaust, lung cancer, occupational exposure. PMID- 15531441 TI - Understanding the spatial clustering of severe acute respiratory syndrome (SARS) in Hong Kong. AB - We applied cartographic and geostatistical methods in analyzing the patterns of disease spread during the 2003 severe acute respiratory syndrome (SARS) outbreak in Hong Kong using geographic information system (GIS) technology. We analyzed an integrated database that contained clinical and personal details on all 1,755 patients confirmed to have SARS from 15 February to 22 June 2003. Elementary mapping of disease occurrences in space and time simultaneously revealed the geographic extent of spread throughout the territory. Statistical surfaces created by the kernel method confirmed that SARS cases were highly clustered and identified distinct disease "hot spots." Contextual analysis of mean and standard deviation of different density classes indicated that the period from day 1 (18 February) through day 16 (6 March) was the prodrome of the epidemic, whereas days 86 (15 May) to 106 (4 June) marked the declining phase of the outbreak. Origin and-destination plots showed the directional bias and radius of spread of superspreading events. Integration of GIS technology into routine field epidemiologic surveillance can offer a real-time quantitative method for identifying and tracking the geospatial spread of infectious diseases, as our experience with SARS has demonstrated. PMID- 15531440 TI - In vivo imaging of activated estrogen receptors in utero by estrogens and bisphenol A. AB - Environmental estrogens are of particular concern when exposure occurs during embryonic development. Although there are good models to study estrogenic activity of chemicals in adult animals, developmental exposure is much more difficult to test. The weak estrogenic activity of the environmental estrogen bisphenol A (BPA) in embryos is controversial. We have recently generated transgenic mice that carry a reporter construct with estrogen-responsive elements coupled to luciferase. We show that, using this in vivo model in combination with the IVIS imaging system, activation of estrogen receptors (ERs) by maternally applied BPA and other estrogens can be detected in living embryos in utero. Eight hours after exposure to 1 mg/kg BPA, ER transactivation could be significantly induced in the embryos. This was more potent than would be estimated from in vitro assays, although its intrinsic activity is still lower than that of diethylstilbestrol and 17beta-estradiol dipropionate. On the basis of these results, we conclude that the estrogenic potency of BPA estimated using in vitro assays might underestimate its estrogenic potential in embryos. PMID- 15531442 TI - Assessing ozone-related health impacts under a changing climate. AB - Climate change may increase the frequency and intensity of ozone episodes in future summers in the United States. However, only recently have models become available that can assess the impact of climate change on O3 concentrations and health effects at regional and local scales that are relevant to adaptive planning. We developed and applied an integrated modeling framework to assess potential O3-related health impacts in future decades under a changing climate. The National Aeronautics and Space Administration-Goddard Institute for Space Studies global climate model at 4 degrees x 5 degrees resolution was linked to the Penn State/National Center for Atmospheric Research Mesoscale Model 5 and the Community Multiscale Air Quality atmospheric chemistry model at 36 km horizontal grid resolution to simulate hourly regional meteorology and O3 in five summers of the 2050s decade across the 31-county New York metropolitan region. We assessed changes in O3-related impacts on summer mortality resulting from climate change alone and with climate change superimposed on changes in O3 precursor emissions and population growth. Considering climate change alone, there was a median 4.5% increase in O3-related acute mortality across the 31 counties. Incorporating O3 precursor emission increases along with climate change yielded similar results. When population growth was factored into the projections, absolute impacts increased substantially. Counties with the highest percent increases in projected O3 mortality spread beyond the urban core into less densely populated suburban counties. This modeling framework provides a potentially useful new tool for assessing the health risks of climate change. PMID- 15531443 TI - Induced sputum assessment in New York City firefighters exposed to World Trade Center dust. AB - New York City Firefighters (FDNY-FFs) were exposed to particulate matter and combustion/pyrolysis products during and after the World Trade Center (WTC) collapse. Ten months after the collapse, induced sputum (IS) samples were obtained from 39 highly exposed FDNY-FFs (caught in the dust cloud during the collapse on 11 September 2001) and compared to controls to determine whether a unique pattern of inflammation and particulate matter deposition, compatible with WTC dust, was present. Control subjects were 12 Tel-Aviv, Israel, firefighters (TA-FFs) and 8 Israeli healthcare workers who were not exposed to WTC dust. All controls volunteered for this study, had never smoked, and did not have respiratory illness. IS was processed by conventional methods. Retrieved cells were differentially counted, and metalloproteinase-9 (MMP-9), particle size distribution (PSD), and mineral composition were measured. Differential cell counts of FDNY-FF IS differed from those of health care worker controls (p < 0.05) but not from those of TA-FFs. Percentages of neutrophils and eosinophils increased with greater intensity of WTC exposure (< 10 workdays or greater than or equal to 10 workdays; neutrophils p = 0.046; eosinophils p = 0.038). MMP-9 levels positively correlated to neutrophil counts (p = 0.002; r = 0.449). Particles were larger and more irregularly shaped in FDNY-FFs (1-50 microm; zinc, mercury, gold, tin, silver) than in TA-FFs (1-10 microm; silica, clays). PSD was similar to that of WTC dust samples. In conclusion, IS from highly exposed FDNY FFs demonstrated inflammation, PSD, and particle composition that was different from nonexposed controls and consistent with WTC dust exposure. PMID- 15531445 TI - Geographic analysis of blood lead levels in New York State children born 1994 1997. AB - We examined the geographic distribution of the blood lead levels (BLLs) of 677,112 children born between 1994 and 1997 in New York State and screened before 2 years of age. Five percent of the children screened had BLLs higher than the current Centers for Disease Control and Prevention action level of 10 microg/dL. Rates were higher in upstate cities than in the New York City area. We modeled the relationship between BLLs and housing and socioeconomic characteristics at the ZIP code level. Older housing stock, a lower proportion of high school graduates, and a higher percentage of births to African-American mothers were the community characteristics most associated with elevated BLLs. Although the prevalence of children with elevated BLLs declined 44% between those born in 1994 and those born in 1997, the rate of improvement may be slowing down. Lead remains an environmental health problem in inner-city neighborhoods, particularly in upstate New York. We identified areas having a high prevalence of children with elevated BLLs. These communities can be targeted for educational and remediation programs. The model locates areas with a higher or lower prevalence of elevated BLLs than expected. These communities can be studied further at the individual level to better characterize the factors that contribute to these differences. PMID- 15531444 TI - Maternal and paternal risk factors for cryptorchidism and hypospadias: a case control study in newborn boys. AB - Little is known on environmental risk factors for cryptorchidism and hypospadias, which are among the most frequent congenital abnormalities. The aim of our study was to identify risk factors for cryptorchidism and hypospadias, with a focus on potential endocrine disruptors in parental diet and occupation. In a case-control study nested within a cohort of 8,698 male births, we compared 78 cryptorchidism cases and 56 hypospadias cases with 313 controls. The participation rate was 85% for cases and 68% for controls. Through interviews, information was collected on pregnancy aspects and personal characteristics, lifestyle, occupation, and dietary phytoestrogen intake of both parents. Occupational exposure to potential endocrine disruptors was classified based on self-reported exposure and ratings of occupational hygienists based on job descriptions. Our findings indicate that paternal pesticide exposure was associated with cryptorchidism [odds ratio (OR) = 3.8; 95% confidence interval (95% CI), 1.1-13.4]. Smoking of the father was associated with hypospadias (OR = 3.8; 95% CI, 1.8-8.2). Maternal occupational, dietary, and lifestyle exposures were not associated with either abnormality. Both abnormalities were associated with suboptimal maternal health, a lower maternal education, and a Turkish origin of the parents. Being small for gestational age was a risk factor for hypospadias, and preterm birth was a risk factor for cryptorchidism. Because paternal pesticide exposure was significantly associated with cryptorchidism and paternal smoking was associated with hypospadias in male offspring, paternal exposure should be included in further studies on cryptorchidism and hypospadias risk factors. PMID- 15531446 TI - The relationship between housing and health: children at risk. AB - In November 2002, the National Center for Healthy Housing convened a 2-day workshop to review the state of knowledge in the field of healthy housing. The workshop, supported with funds from the U.S. Centers for Disease Control and Prevention's National Center for Injury Prevention and Control and National Center for Environmental Health, was unique in that it focused solely on the effect of housing on children's health and the translation of research findings into practical activities in home construction, rehabilitation, and maintenance. Participants included experts and practitioners representing the health, housing, and environmental arenas. Presentations by subject-matter experts covered four key areas: asthma, neurotoxicants, injury, and translational research. Panel discussions followed the presentations, which generated robust dialogue on potential future research opportunities and overall policy gaps. Lack of consensus on standard measurements, incomplete understanding about the interaction of home hazards, inadequate research on the effectiveness of interventions, and insufficient political support limit current efforts to achieve healthy housing. However, change is forthcoming and achievable. Key words: asthma, childhood exposure, environmental toxicants, healthy housing, lead poisoning. PMID- 15531447 TI - Cytoreductive therapy for patients with essential thrombocythemia at high risk of thromboembolic complications. The difficult choice of the optimal drug. PMID- 15531448 TI - Primary cutaneous lymphoma: the witness of multidisciplinarity. PMID- 15531449 TI - Risk of deep vein thrombosis: interaction between oral contraceptives and high factor VIII levels. PMID- 15531450 TI - In vivo reduction of erythrocyte oxidant stress in a murine model of beta thalassemia. AB - BACKGROUND AND OBJECTIVES: Oxidant damage is an important contributor to the premature destruction of erythrocytes and anemia in thalassemias. To assess the extent of oxidant damage of circulating erythrocytes and the effects of antioxidant therapy on erythrocyte characteristics and anemia, we used a mouse model of human beta-thalassemia intermedia (b1/b2 deletion). DESIGN AND METHODS: Several parameters indicative of oxidant damage were measured at baseline and following administration of the semi-synthetic flavonoid antioxidant, 7 monohydroxyethylrutoside (monoHER), to beta-thalassemic mice at a dose of either 500 mg/kg i.p. once a day (n=6) or 250 mg/kg i.p. twice a day (n=6) for 21 days. RESULTS: Significant erythrocyte oxidant damage at baseline was indicated by: (i) dehydration, reduced cell K content, and up-regulated K-Cl co-transport; (ii) marked membrane externalization of phosphatidylserine; (iii) reduced plasma and membrane content of vitamin E; and (iv) increased membrane bound IgG. MonoHER treatment increased erythrocyte K content, and markedly improved all cellular indicators of oxidant stress and of lipid membrane peroxidation. While anemia did not improve, monoHER therapy reduced reticulocyte counts, improved survival of a fraction of red cells, and reduced ineffective erythropoiesis with decreased total bilirubin, lactate dehydrogenase and plasma iron. INTERPRETATION AND CONCLUSIONS: Antioxidant therapy reverses several indicators of oxidant damage in vivo. These promising antioxidant effects of monoHER should be investigated further. PMID- 15531451 TI - Hemochromatosis gene mutations and iron metabolism in celiac disease. AB - BACKGROUND AND OBJECTIVES: Iron deficiency anemia is a common manifestation of celiac disease, which may be due to genetic and environmental factors. HFE mutations, frequent in Caucasian populations, can cause increased intestinal iron absorption and thus could protect against the development of iron deficiency. The aim of this study was to evaluate the prevalence of HFE mutations and their effect on iron metabolism in Italian celiac patients at diagnosis and after a gluten-free diet. DESIGN AND METHODS: C282Y and H63D mutations were assessed by polymerase chain reaction (PCR) and restriction enzyme digestion in 203 patients with celiac disease and in 206 controls. HLA alleles were determined by sequence specific primers and PCR. Duodenal histology was graded using Marsh's classification, and iron parameters measured by standard techniques. RESULTS: The frequency of the C282Y mutation was similar in celiac patients and controls (0.034 vs. 0.031); comparable frequencies were detected also for the H63D allele (0.170 vs. 0.136 in celiac patients and controls, respectively). Neither of the two HFE mutations affected iron indices in celiac patients at diagnosis, whereas a significant inverse correlation was detected between hemoglobin or ferritin and severity of histological damage (Marsh 3C or 3B vs. 3A, p<0.05 for both parameters). After a gluten-free diet, a slight increase in hemoglobin levels was observed in C282Y carriers as compared to controls, but only in female patients (p=0.044). INTERPRETATION AND CONCLUSIONS: In Italian patients with untreated celiac disease, HFE mutations do not constitute a protective factor against the development of iron deficiency, which seems to be mainly determined by the severity of the intestinal lesions. PMID- 15531452 TI - A long-term study of young patients with essential thrombocythemia treated with anagrelide. AB - BACKGROUND AND OBJECTIVES: Essential thrombocythemia (ET) can be complicated by life-threatening thrombosis and has a risk of converting into acute leukemia. Cytoreductive therapy may reduce the risk of thromboembolic complications. Herein, we report the results of a long-term study of patients with ET treated with anagrelide to control thrombocytosis. DESIGN AND METHODS: Thirty-nine (34 evaluable) patients (median age, 33 years; 24 previously untreated) were enrolled between 1989-1996; the mean platelet count prior to therapy was 1197x10(9)/L. Only 9 out of 34 evaluable patients were at high risk of thrombosis (platelet count more than 1500x10(9)/L). The initial dose of anagrelide (0.5 mg/bid for 7 days) was increased by 0.5 mg/day (maximum dose: 3 mg/day) until a response was seen. RESULTS: A complete response (platelets < 450x10(9)/L for >1 month) was seen in 15 /34 (44%) patients and a partial response (platelets 450-600x10(9)/L for >1 month) was seen in 17/34 ( 50%), so that the some kind of response was seen in 32/34 (94%) of the patients at a median time of 4.2 months after starting treatment. Seventeen patients (50%) are still being treated and have achieved platelet control for a maximum follow-up of 12.5 years. Late onset anemia occurred in 4/39 patients. Three out of 39 patients (8%) had cardiac disorders. INTERPRETATION AND CONCLUSIONS: Anagrelide appears suitable for controlling thrombocytosis in ET patients over the long-term. This drug may be used in patients younger than 60 years, with the exclusion of women of child-bearing potential and subjects aged 40-60 years with a history of major thrombotic events. Anagrelide should not be administered to patients with cardiac disorders, and a careful approach to patients should include monitoring of heart function before and during treatment. PMID- 15531453 TI - The granzyme B inhibitor PI-9 is differentially expressed in all main subtypes of pediatric acute lymphoblastic leukemias. AB - BACKGROUND AND OBJECTIVES: The success of bone marrow transplantation in leukemia depends on graft-versus-leukemia (GvL) effects, mediated by cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. These act by CD95L or granule cytotoxins, such as granzyme B (GrB) whose only known inhibitor is proteinase inhibitor 9 (PI-9). Since PI-9 protects cells from CTL, PI-9 may counteract GvL in leukemias. Our aim was to establish methods to analyze the expression and function of PI-9. DESIGN AND METHODS: We screened the most common pediatric ALL subsets, i.e. pre-B, common, and cortical T-ALL, for PI-9 expression, using a reverse transcription polymerase chain reaction (RT-PCR) approach which was correlated to semiquantitative and functional methods established in cell lines and patient probes. RESULTS: In vitro, PI-9 mediated resistance towards CTL and inhibited GrB-induced clevage of caspase 3. In patient-derived ALL cells, PI-9 high and PI-9 low specimens were studied by flow cytometry, RT-PCR, in vitro GrB treatment and NK assay, demonstrating concordant results and PI-9-dependent target cell protection. Analysis of PI-9 in probes from ALL patients showed differential expression, but no correlation with immunotype. INTERPRETATION AND CONCLUSIONS: Our data suggest that PI-9 in pediatric ALL is differentially expressed, without close correlation to subtype. Since PI-9 considerably alters GrzB and killer cell sensitivity, it may strongly influence the efficacy of GvL effects. The approaches applied here will allow evaluation of the expression and function of PI-9 in larger series of malignancies. PMID- 15531454 TI - Frequent polymorphic changes but not mutations of TRAIL receptors DR4 and DR5 in mantle cell lymphoma and other B-cell lymphoid neoplasms. AB - BACKGROUND AND OBJECTIVES: Tumor necrosis factor related apoptosis-inducing ligand (TRAIL) receptors DR4 and DR5 have been mapped to chromosome 8p21-22, a region frequently deleted in different lymphoid neoplasms. DESIGN AND METHODS: To investigate the potential alterations of these genes in lymphoid neoplasms, we examined the presence of gene mutations in exons 3, 4, and 9 in 69 cases with mantle cell lymphoma (MCL), 16 with chronic lymphocytic leukemia (CLL), 12 with follicular lymphomas (FL) and 17 with large B-cell-lymphomas (DLBCL), as well as in 4 lymphoid cell lines carrying the t(11;14) translocation, and 91 healthy blood donors. RESULTS: Three CLL and three MCL cases had 8p deletions. Two nucleotide changes in or near the intron 3 splice consensus sequence and a silent change were found. These rare changes were also present in the germ-line of the patients. The DR4 death domain A1322G polymorphism was significantly more frequent in MCL [odds ratio (OR) = 5.9; 95% confidence interval (CI), 1.92-18.1] and CLL (OR = 4.5; CI, 1.18-17) patients than in a sex and age-adjusted healthy population. In contrast, the DR4 exon 4 C626G polymorphism was associated with a significant overall decreased risk for MCL (OR = 0.3; CI, 0.12-0.8). No mutations or cancer-associated polymorphic changes were found in DR5 domains. INTERPRETATION AND CONCLUSIONS: These findings indicate that mutations of DR4 and DR5 are uncommon in lymphoid neoplasms but DR4 polymorphic alleles may contribute to the pathogenesis of these malignancies. PMID- 15531455 TI - Molecular genetic analysis of severe coagulation factor XI deficiency in six Italian patients. AB - BACKGROUND AND OBJECTIVES: Factor XI (FXI) deficiency is a rare autosomal recessive coagulopathy which is, however, frequent among Ashkenazi Jews. Two mutations, type II (Glu117stop) and type III (Phe283Leu), account for the majority of abnormal alleles in this population. The aim of this study was to analyze the molecular basis of FXI deficiency in six unrelated Italian probands with severe deficiency, a population hitherto largely unexplored. DESIGN AND METHODS: All patients showed unmeasurable functional FXI levels in plasma. Mutational screening was performed by sequencing. Haplotype analysis was performed using intragenic polymorphisms. Expression studies were performed by transient transfection in COS-1 cells. RESULTS: Sequencing identified two novel mutations: a nonsense mutation (Cys118stop) in exon 5 in two probands, and a 6-bp deletion (643-648delATCGAC) in exon 7 in one proband. The Cys118stop is predicted to cause FXI deficiency by a secretion defect and/or by increased mRNA degradation. The 6-bp deletion causes the loss of residues Ile197 and Asp198. There was a remarkable secretion impairment of the deleted FXI protein. In four of the six probands, the type II mutation was found. Haplotype analysis in patients carrying the type II mutation revealed that they share a common haplotype, perhaps derived from a Jewish ancestor. INTERPRETATION AND CONCLUSIONS: The identification and characterization of two novel mutations widen the mutational spectrum of FXI deficiency. Haplotype analysis is compatible with a Jewish origin of the type II mutation. The high occurrence of the type II mutation among Italian patients will be helpful to direct future genetic screenings. PMID- 15531456 TI - High prevalence of acquired von Willebrand's syndrome in patients with thyroid diseases undergoing thyroid surgery. AB - BACKGROUND AND OBJECTIVES: Various coagulation abnormalities occur in patients with thyroid diseases. These abnormalities range from subclinical laboratory findings to hemorrhage or thromboembolism. However, the prevalence of hemostatic abnormalities in patients with thyroid diseases is still unclear. DESIGN AND METHODS: Between January 1999 and December 2003, 1342 consecutive patients with various thyroid diseases who were candidates for thyroid surgery underwent preoperative screening of hemostatic parameters including prothrombin time, activated partial thromboplastin time and platelet-related hemostasis with the PFA-100 platelet-function analyzer. RESULTS: Thirty-nine patients (2.9%) had abnormalities of the coagulation screening tests. Of these, 35 patients had von Willebrand's disease (type 1 in 33 cases and type 2A in 2 cases), 2 patients had decreased platelet aggregability, and 2 patients had coagulation factor XI deficiency. As all patients with coagulation abnormalities responded to subcutaneous desmopressin injection (0.3 microg/Kg BW), this drug was successfully used as surgical prophylaxis. INTERPRETATION AND CONCLUSIONS: Up to 3% of patients with thyroid diseases undergoing thyroid surgery have coagulation abnormalities, in most cases resembling von Willebrand's disease. Coagulation screening tests are needed in order to identify those patients at increased risk of bleeding. PMID- 15531457 TI - Risk of deep vein thrombosis: interaction between oral contraceptives and high factor VIII levels. AB - BACKGROUND AND OBJECTIVES: High levels of factor VIII are a common risk factor for venous thromboembolism (VTE). The aim of this study was to evaluate the possible interaction between the presence of high factor VIII levels and oral contraceptive (OC) use. DESIGN AND METHODS: Factor VIII levels were measured by a chromogenic assay in 174 women who suffered from VTE in reproductive age and in 484 healthy women. Subjects carrying thrombophilic alterations were excluded. RESULTS: Factor VIII levels were stratified into quartiles. The adjusted odds ratio (OR) of VTE in subjects with factor VIII levels in the upper quartile (>1.95 IU/mL) was 7.45 (95%CI: 3.80-14.6). Among the 174 patients, 85 had experienced VTE during OC use. The 179 healthy women who had used OC for at least 6 months in the two years before presentation but had stopped OC at least 3 months before the blood sampling were considered as OC users. The risk of VTE among OC users with factor VIII levels in the upper quartile was increased about 13-fold (OR: 13.0, 95% CI: 4.92-34.3). INTERPRETATION AND CONCLUSIONS: Our results showed that there is an increased risk of VTE due to oral contraceptive use in women with elevated factor VIII and, as has been previously described for factor V Leiden and G20210A prothrombin mutations, the raised level of the coagulation factor and oral contraceptive use seem to have a synergistic effect. PMID- 15531458 TI - Infliximab treatment for steroid-refractory acute graft-versus-host disease. AB - BACKGROUND AND OBJECTIVES: Tumor necrosis factor a is one of the principal cytokines involved in the pathogenesis of acute graft-versus-host-disease (GVHD). Infliximab is an antibody to this cytokine. DESIGN AND METHODS: We performed a retrospective analysis to evaluate the activity of infliximab in 32 patients with severe steroid-refractory acute GVHD. The patients received a median of 3 weekly courses of infliximab. The main organs involved in the patients were skin (n=2) liver (n=1), bowel (n=19), liver and bowel at the same stage (n=10). RESULTS: Nineteen out 32 patients (59%) responded to infliximab with 6 (19%) complete and 13 (40%) partial responses. Age younger than 35 years, intestinal involvement and a longer time between hematopoietic stem cell transplantation and infliximab administration were factors predicting a favorable response. Infective episodes developed in 23/32 (72%) patients. All the 13 unresponsive patients died of GVHD shortly after infliximab. Thirteen of 19 responsive patients were alive at a median follow-up of 449 days (range 155-842) after infliximab, with no signs of chronic GVHD (n=5), limited (n=5) or extensive involvement (n=3). Six patients who responded subsequently died, one of chronic lung GVHD, the others of vascular complications or infections (2 fungal diseases). INTERPRETATION AND CONCLUSIONS: We conclude that infliximab is active in the treatment of severe steroid refractory acute GVHD, particularly when the intestine is involved. Infections commonly followed its administration. The clinical activity of infliximab and the possibility that it increases the risk of infections are worth investigating in prospective trials. PMID- 15531459 TI - Primary cutaneous B-cell lymphomas. Current concepts. I. AB - BACKGROUND: Primary cutaneous B-cell lymphoma (pCBCL) has only recently been recognized as a distinct clinicopathologic entity. This entity represents a wide spectrum of lymphoproliferative disorders that must be separated from non Hodgkin's B-cell lymphomas secondarily involving the skin and cutaneous B-cell pseudolymphomas. They are defined as B-cell lymphomas originating in the skin, with no evidence of extracutaneous disease at presentation, as assessed by adequate staging procedures. INFORMATION SOURCES: With the advent of improved immunophenotyping and immunogenotyping, increasing numbers of pCBCL cases are being diagnosed. However, there is still confusion regarding the classification, treatment, and prognosis of these patients. The aim of this paper is to provide the clinician with a concise summary of the diagnosis, course, and treatment of pCBCL. Currently, the European Organization for Research and Treatment of Cancer classification is the most adequate classification scheme, identifying the subtypes of pCBCL by clinical behavior and histopathologic findings, and allowing a better management of the patients. STATE OF THE ART: Based on this classification, the most common subtypes of pCBCL are follicular center cell lymphoma and marginal zone B-cell lymphoma, indolent lymphomas with an excellent prognosis (>95% 5-year survival rate in our series). Although local cutaneous recurrences are observed in about 25% of patients, dissemination to internal organs is rare. A less common, specific subtype - the so-called large B-cell lymphoma of the leg(s) - generally has a poorer prognosis, with a 5-year survival rate of approximately 60%. PERSPECTIVES: As a rule, pCBCL is highly responsive to radiation therapy, which should be considered the treatment of choice. Polychemotherapy should be restricted to patients with involvement of several noncontiguous anatomic sites, those refractory or plurirelapsed after radiotherapy, or those with extracutaneous spread. PMID- 15531460 TI - Cutaneous T-cell lymphomas (including rare subtypes). Current concepts. II. AB - BACKGROUND: Cutaneous T-cell lymphomas (CTCL) represent about the 80% of skin lymphomas and comprise a heterogeneous group of diseases with respect to clinical presentation, outcome, histologic and immunophenotypic features. In the recent years, data have been accumulated indicating that clinical and biological differences exists between primary nodal and primary extranodal lymphomas. INFORMATION SOURCES: On such bases, the cutaneous lymphoma classification by the EORTC (European Organization for Cancer Research and Treatment) but also the recent general lymphoma classification, by the WHO (World Health Organization), have dedicated special attentions to the cutaneous lymphomas including the T and NK-cell subtypes. This paper reviews the most significative subtypes of T-cell lymphoma that affect the skin primarily or secondarily. STATE OF THE ART: Apart from mycosis fungoides with its variants forms and the Sezary's syndrome, we have focused on the CD30+ primary cutaneous lymphoproliferative disorders (PCLD) (25% of all CTCL), a fascinating spectrum of disease, extending from lymphomatoid papulosis (LyP) trough to CD30+ large cell lymphoma. These disorders have in common large atypical CD30+ cells and a frequent spontaneous regression of the skin lesions associated with a relatively favourable outcome (excellent in LyP). The identification of this group of skin disorders is crucial for the patients since most of CD30+ PCLD are indolent diseases that do not warrant aggressive treatment. Others types of CTCL include the heterogeneous category of peripheral T-cell lymphoma not otherwise specified (NOS), subcutaneous panniculitis-like T cell lymphoma and the still controversial group of the cytotoxic lymphomas. PERSPECTIVES: Notably, the latter two subtypes have special relevance to the clinicians because (i) subcutaneous panniculitis-like T-cell lymphoma may be associated with the hemophagocytic syndrome; (ii) skin lesions in cytotoxic lymphomas may represent the first manifestation of an otherwise systemic disease. PMID- 15531461 TI - A pilot study on the efficacy of ketorolac plus tramadol infusion combined with erythrocytapheresis in the management of acute severe vaso-occlusive crises and sickle cell pain. AB - One of the major causes of hospitalization for patients with sickle cell disease (SCD) are vaso-occlusive crises (VOC), which are characterized by acute pain and organ damage related to the presence of dense red cells. Here we report a pilot study which combined balanced analgesia with tramadol plus ketorolac and erythrocytapheresis. Key words: sickle cell disease, therapeutic erythrocytapheresis, HbS, visual analog scale, vaso-occlusive crisis. PMID- 15531462 TI - Elevated thrombopoietin levels and alterations in the sequence of its receptor, c Mpl, in patients with Diamond-Blackfan anemia. AB - In the study of the possible thrombopoietin (TPO)-c-Mpl pathway involvement in the pathogenesis of Diamond-Blackfan anemia, repeatedly increased serum TPO levels were identified in 7/14 patients and changes in c-mpl sequence in 3/14 patients. While elevated TPO levels can represent a compensatory mechanism for impaired erythropoiesis, c-Mpl mutations could influence the disease severity. PMID- 15531463 TI - Latency of onset of de novo myelodysplastic syndromes. AB - The latency of onset of de novo myelodysplastic syndromes (MDS) is unknown. We report a retrospective analysis of blood counts from patients with MDS and acute myeloid leukemia (AML), and demonstrate temporal differences in rates of change of hemoglobin concentration and mean cell volume within 2-3 years of diagnosis, indicative of the earliest evidence of disease. PMID- 15531464 TI - Anagrelide-associated cardiomyopathy in polycythemia vera and essential thrombocythemia. AB - A comprehensive database inquiry at our institutions identified 11 patients with echocardiogram-documented idiopathic cardiomyopathy that post-dated a diagnosis of either polycythemia vera or essential thrombocythemia. Anagrelide therapy was temporally associated with the particular complication in 6 patients, all of whom experienced symptomatic and/or objective improvement after drug discontinuation. PMID- 15531465 TI - ABC transporter ABCA3 is expressed in acute myeloid leukemia blast cells and participates in vesicular transport. AB - Drug resistance is a major issue in the treatment of acute myeloid leukemia (AML), and drug efflux by ATP-binding-cassette (ABC) transporters is one of the main mechanism involved in this resistance. We determined the prevalence of the intracellular transporter ABCA3 in specimens from patients with AML, and addressed its biology with attention to intracellular compartmentalization. PMID- 15531466 TI - Prognostic significance of N-RAS and K-RAS mutations in 232 patients with acute myeloid leukemia. AB - Reports on the prognostic impact of mutations in the RAS proto-oncogenes in patients with acute myeloid leukemia (AML) are conflicting. A peptide nucleic acid (PNA)-based technique was used on 232 AML samples to detect point mutations of the hotspots in N-RAS and K-RAS. No significant correlations between RAS mutations and clinical features, karyotype or FLT3 were found. PMID- 15531467 TI - Overexpression of Her2/neu is observed in one third of adult acute lymphoblastic leukemia patients and is associated with chemoresistance in these patients. AB - Among 100 patients with acute lymphoblastic leukemia (ALL), 15 B-ALL patients were found positive for surface Her2/neu expression. The incidence in children was only 3.4% compared to 31% in adults (p=0.001). Considering only adult B-ALL patients (n=38), surface Her2/neu expression was associated with chemoresistance (50% versus 11%, p=0.03) suggesting that it could be a prognostic marker of poor clinical outcome in ALL. PMID- 15531468 TI - Methylation status of the p15, p16 and MGMT promoter genes in primary cutaneous T cell lymphomas. AB - p15(INK4b), p16(INK4a) and O(6)-methylguanine DNA methyltransferase (MGMT) gene hypermethylation was studied in 22 patients with primary cutaneous T-cell lymphomas (CTCL). p15(INK4b) and p16(INK4a) inactivation is present in early and advanced disease and seems to be independent of disease stage. MGMT inactivation may play a pathogenetic role in a subset of CTCL. PMID- 15531469 TI - Longitudinal variation of hemoglobin and reticulocytes in elite rowers. AB - Longitudinal monitoring of athlete's hematologic parameters holds considerable promise as a strategy to detect and thereby deter illicit blood doping. This study documents temporal changes of hemoglobin concentration (Hb) and reticulocyte counts in elite rowers. The 'within subject' variation in rowers was comparable to that of athletes from other sports. Reticulocyte results were dependent on the type of instrument used. PMID- 15531471 TI - Monitoring the response to treatment in acromegaly. PMID- 15531472 TI - Intelligence and psychosocial functioning during long-term growth hormone therapy in children born small for gestational age. PMID- 15531473 TI - Intelligence and psychosocial functioning during long-term growth hormone therapy in children born small for gestational age. AB - Short stature is not the only problem faced by small for gestational age (SGA) children. Being born SGA has also been associated with lowered intelligence, poor academic performance, low social competence, and behavioral problems. Although GH treatment in short children born SGA can result in a normalization of height during childhood, the effect of GH treatment on intelligence and psychosocial functioning remains to be investigated. We show the longitudinal results of a randomized, double-blind, GH-dose response study initiated in 1991 to follow growth, intelligence quotient (IQ), and psychosocial functioning in SGA children during long-term GH treatment. Patients were assigned to one of two treatment groups (1 or 2 mg GH/m(2) body surface.d, or approximately 0.035 or 0.07 mg/kg.d). Intelligence and psychosocial functioning were evaluated at start of GH treatment (n = 74), after 2 yr of GH treatment (n = 76), and in 2001 (n = 53). IQ was assessed by a short-form Wechsler Intelligence Scale for Children-Revised or Wechsler Adult Intelligence Scale (Block-design and Vocabulary subtests). Behavioral problems were measured by the Achenbach Child Behavior Checklist or Young Adult Behavior Checklist, and self-perception was measured by the Harter Self-Perception Profile. Mean (sem) birth length sd score was -3.6 (0.2), mean age and height at start was 7.4 (0.2) yr and -3.0 (0.1) sd score, respectively, mean duration of GH treatment was 8.0 (0.2) yr, and mean age in 2001 was 16.5 (0.3) yr. After 2 yr of GH treatment, 96% of both GH groups showed a height gain sd score of 1 sd from the start of treatment or more, resulting in a normal height (i.e. height >/= -2.0 sd for age and sex) in 70% of the children. In 2001, 48 (91%) of the 53 children participating in this study had reached a normal height. Block-design s-score and the estimated total IQ significantly increased (P < 0.001 and P < 0.001, respectively) from scores significantly lower than Dutch peers at start (P < 0.001 and P < 0.001, respectively) to comparable scores in 2001. The increase over time for the Vocabulary s-score was not significant. Internalizing Behavior sd scores remained comparable to Dutch peers, whereas Externalizing Behavior sd scores and Total Problem Behavior sd scores improved significantly during GH therapy (P < 0.01 and P < 0.05, respectively) to scores comparable to Dutch peers. Self-perception sd scores improved from start of GH treatment until 2001 (P < 0.001) to scores significantly higher than Dutch peers (P < 0.05). No significant differences between the two GH dosage groups were found. Improvement in Externalizing and Total Problem Behavior sd scores over time was significantly related to change in height sd score (P < 0.05 and P < 0.01, respectively), whereas scores over time for Vocabulary, Block-design, Internalizing, or total Harter Self-Perception score were not related to change in height sd scores. In conclusion, parallel to a GH-induced catch-up growth in adolescents born SGA, IQ, behavior, and self-perception showed a significant improvement over time from scores below average to scores comparable to Dutch peers. In addition, children whose height over time became closer to that of their peers showed less problem behavior. PMID- 15531474 TI - Choroidal and skin metastases from papillary thyroid cancer: case and a review of the literature. AB - A patient with widely metastatic papillary thyroid cancer who had been previously treated with (131)I and external beam radiation presented with purple nodular lesions on his face and scalp. On biopsy, the nodules were papillary carcinoma with cells that stained for thyroglobulin. Subsequently he developed decreased left eye visual acuity, and fundoscopy revealed lesions typical of choroidal metastases. Dermal and choroidal metastases of papillary thyroid carcinoma are both rare. However, the significance of these clinical manifestations may be overlooked and ignored unless the diagnosis is considered. New skin nodules or visual acuity decline in a patient with papillary thyroid cancer may represent manifestations of distant metastatic disease and should prompt thorough evaluation with dermatological examination and fundoscopy. Choroidal and skin metastases have almost always occurred in patients with advanced disease, but initial presentation with these lesions is possible, and in such instances a thorough search for additional sites of metastatic disease is recommended. Occasionally such metastases may respond to (131)I therapy or external beam radiation. PMID- 15531475 TI - Long-term outcome of patients with acromegaly and congestive heart failure. AB - Cardiovascular complications are a major cause of morbidity and mortality in patients with acromegaly. Normalization of GH secretion is associated with an improvement in structural and functional cardiac abnormalities. However, the long term cardiac effects of treatment for acromegaly have not been studied in patients who have already developed chronic congestive heart failure (CHF). We reviewed the charts of 330 consecutive patients with acromegaly treated in two French and Belgian centers since 1985. Ten patients with both acromegaly and CHF (eight men, two women, mean age 49.7 yr) were studied retrospectively. One of them was excluded because CHF was due to severe aortic stenosis.CHF (New York Heart Association stages III-IV and echocardiography showing dilated hypokinetic cardiomyopathy with left ventricular systolic dysfunction and a left ventricular ejection fraction less than 45%) was diagnosed before, concomitantly, or after acromegaly in, respectively, two, five, and two patients. Three patients were referred with terminal heart failure requiring transplantation.One patient had transient CHF associated with a hypertensive crisis. The other eight patients had symptomatic chronic CHF. Control of GH hypersecretion failed, totally or partially, in three patients: one had a long-term survival, and the two others died at 1 and 5 yr. Good GH control was achieved in five patients: four of these are still alive 2-16 yr after diagnosis of CHF, their clinical status is stable or improved, and their quality of life is good. Overall, the 1- and 5-yr mortality (or transplantation) rates for patients with chronic symptomatic CHF were 25% (2 of 8 patients) and 37.5% (3 of 8 patients), respectively. In conclusion, less than 3% of acromegalic patients developed CHF in this study. Although effective treatment of acromegaly improved short-term cardiovascular status, its impact on long-term survival is questionable. PMID- 15531476 TI - Developmental trends in cord and postpartum serum thyroid hormones in preterm infants. AB - The purpose of this study was first to clarify postnatal trends in sera T(4), free T(4) (FT(4)), T(4)-binding globulin, TSH, T(3), rT(3), and T(4) sulfate levels in cord and at 7, 14, and 28 d in groups of preterm infants at 23-27 wk (n = 101), 28-30 wk (n = 196), and 31-34 (n = 253) wk gestation, and second to compare these trends to those of term infants and also with cord sera levels of equivalent gestational ages (n = 812; 23-42 wk gestation). In all preterm groups, TSH and rT(3) decrease to below, T(4)-binding globulin increases to within, and T(3) and T(4) sulfate increase to above cord levels of equivalent gestational age. Term infants are hyperthyroxinemic relative to cord and nonpregnant adult levels of T(4). Postnatal T(4) increases are attenuated in 31- to 34-wk infants, absent in 28- to 30-wk infants (although levels are equivalent to gestational age), and crucially reversed in 23- to 27-wk infants. This immature group is hypothyroxinemic relative to other groups and to cord levels of equivalent gestational age. Compared with term infants, postnatal FT(4) increases are lower in 31- to 34-wk infants, attenuated in 28- to 30-wk infants, and absent in 23- to 27-wk infants. The 23- to 27-wk group is distinctive; they are hypothyroxinemic on T(4) levels, yet FT(4) levels are within the cord levels of equivalent gestational age. PMID- 15531477 TI - Stockpiling of transitional and classic primary follicles in ovaries of women with polycystic ovary syndrome. AB - Recently, we proposed an oocyte-growth differentiation factor-9 hypothesis that predicts alterations in the initial stages of folliculogenesis in polycystic ovary syndrome (PCOS) ovaries. Here, we test this hypothesis by scoring the composition of follicles in normal and PCOS ovaries. Follicles were classified as primordial, transitional primary, classic primary, secondary, and Graafian. A total of 2274 follicles were scored. The total number of growing follicles was significantly greater in PCOS ovaries than normal, but the number of nongrowing primordial follicles did not differ. Consequently, the increase in growing follicles in PCOS cannot be explained by increased primordial follicle recruitment. Differential counts showed that the number of growing follicles at each stage of development was significantly greater: PCOS had 2.7-fold more primary, 1.8-fold more secondary, and 2-fold more Graafian follicles than normal. The greatest effect was on the classic primary follicles where the number was almost 5-fold greater in PCOS ovaries. The absence of apoptosis in normal and PCOS preantral follicles argues that the increase in growing follicles in PCOS cannot be explained by changes in atresia. We conclude, therefore, that primary follicle growth is abnormally slow in PCOS and the dynamics are reflected in a stockpiling of classic primary follicles. PMID- 15531478 TI - Cutaneous tumors in patients with multiple endocrine neoplasm type 1 (MEN1) and gastrinomas: prospective study of frequency and development of criteria with high sensitivity and specificity for MEN1. AB - Multiple endocrine neoplasm type 1 (MEN1) is associated with parathyroid, pancreatic, and pituitary tumors. Although most patients present with hyperparathyroidism, the diagnosis can be difficult, because a significant proportion present with other endocrinopathies or may lack a family history, and other MEN1 manifestations may be mild. Recently, multiple cutaneous lesions (angiofibromas and collagenomas) were reported to be frequent in MEN1 patients, and it was proposed that their discovery suggested the diagnosis of MEN1. The purpose of this study was to prospectively assess the frequency and sensitivity/specificity of various cutaneous criteria for MEN1 in 110 consecutive patients with gastrinomas with or without MEN1. All patients had hormonal and functional studies to determine MEN1 status (48 with MEN1, 62 without MEN1), dermatological evaluation, and tumor imaging studies. Angiofibromas and collagenomas were more frequent in MEN1 patients (64% vs. 8% and 62% vs. 5%; P < 0.00001) and were multiple in 77-81% of the MEN1 patients. Lipomas occurred in 17%. The presence of these skin lesions did not correlate with age, disease duration, or other MEN1 features. Angiofibromas or collagenomas (single or multiple) had 50-65% sensitivity for MEN1 and 92-100% specificity. The combination criterion of multiple angiofibromas (more than three) and any collagenomas had the highest sensitivity (75%) and specificity (95%). This criterion has greater sensitivity than pituitary or adrenal disease and is comparable to hyperparathyroidism in some studies of patients with MEN1 with gastrinoma. This criterion should have sufficient sensitivity/specificity to be clinically useful. PMID- 15531479 TI - Patients with familial partial lipodystrophy of the Dunnigan type due to a LMNA R482W mutation show muscular and cardiac abnormalities. AB - Diseases due to mutations in the lamin A/C gene (LMNA) are highly heterogeneous, including neuromuscular and cardiac dystrophies, lipodystrophies, and premature ageing syndromes. In this study we characterized the neuromuscular and cardiac phenotypes of patients bearing the heterozygous LMNA R482W mutation, which is the most frequent genotype associated with the familial partial lipodystrophy of the Dunnigan type (FPLD). Fourteen patients from two unrelated families, including 10 affected subjects, were studied. The two probands had been referred for lipoatrophy and/or diabetes. Lipodystrophy, exclusively observed in LMNA-mutated patients, was of variable severity and limited to postpubertal subjects. Lipodystrophy and metabolic disturbances were more severe in women, even if an enlarged neck was a constant finding. The severity of hypertriglyceridemia and hirsutism in females was related to that of insulin resistance. Clinical muscular alterations were only present in LMNA-mutated patients. Clinical and histological examination showed an invalidating, progressive limb-girdle muscular dystrophy in a 42-yr-old woman that had been present since childhood, associated with a typical postpubertal FPLD phenotype. Six of eight adults presented the association of calf hypertrophy, perihumeral muscular atrophy, and a rolling gait due to proximal lower limb weakness. Muscular histology was compatible with muscular dystrophy in one of them and/or showed a nonspecific excess of lipid droplets (in three cases). Immunostaining of lamin A/C was normal in the six muscular biopsies. Surprisingly, calpain 3 expression was undetectable in the patient with the severe limb-girdle muscular dystrophy, although the gene did not reveal any molecular alterations. At the cardiac level, cardiac septal hypertrophy and atherosclerosis were frequent in FPLD patients. In addition, a 24 yr-old FPLD patient had a symptomatic second degree atrioventricular block. In conclusion, we showed that most lipodystrophic patients affected by the FPLD linked LMNA R482W mutation show muscular and cardiac abnormalities. The occurrence and severity of the myopathic and lipoatrophic phenotypes varied and were not related. The muscular phenotype was evocative of limb girdle muscular dystrophy. Cardiac hypertrophy and advanced atherosclerosis were frequent. FPLD patients should receive careful neuromuscular and cardiac examination whatever the underlying LMNA mutation. PMID- 15531480 TI - Intrafamilial variability of the deafness and goiter phenotype in Pendred syndrome caused by a T416P mutation in the SLC26A4 gene. AB - Pendred syndrome (PS) is the most common cause of syndromic deafness, accounting for more than 5% of all autosomal-recessive hearing loss cases. It is characterized by bilateral sensorineural hearing loss and by goiter with or without hypothyroidism. Mutations in the SLC26A4 gene cause both classical PS and deafness associated with an enlarged vestibular aqueduct without goiter. To investigate a possible genotype-phenotype correlation in PS, we performed a detailed clinical and genetic study in three adult German sibs with typical PS caused by a common homozygous SLC26A4 mutation, T416P. An audiological long-term follow-up of 23 yr showed that the mutation T416P is associated with a distinct type of hearing loss in each of the three sibs: moderate-to-profound progressive deafness, profound nonprogressive deafness, and a milder but more rapidly progressing form. We show that these phenotypic differences are not caused by either different degrees of inner ear malformations or sequence variations in the GJB2/connexin 26 gene. Because the thyroid phenotype was also highly variable within the family, with thyroid sizes ranging from normal to large goiters requiring thyroidectomy, this study leads to the conclusion that other environmental and/or genetic factors have an impact on the PS phenotype. PMID- 15531481 TI - Urocortin 1 in colonic mucosa in patients with ulcerative colitis. AB - Ulcerative colitis (UC) is characterized by a long-standing chronic inflammation of the bowel with intermittent periods of exacerbation and remission. Its acute exacerbation appears to be related to various stresses. Urocortin 1 (Ucn1) may play important roles in integrated local responses to stress. We therefore examined local production of Ucn1 in patients with UC by immunohistochemistry and mRNA in situ hybridization. Ucn1 immunoreactivity was predominantly detected in lamina propria plasma cells and enterochromaffin cells. In UC patients without glucocorticoid treatment, Ucn1-positive cells and plasma cells increased in proportion to the severity of inflammation (P < 0.0001). Ucn1-positive cells significantly increased in UC patients with advanced inflammatory grades, compared with a control group (P < 0.0001) and nonspecific colitis group (P < 0.0001). In glucocorticoid-treated patients, Ucn1-positive cells were significantly lower in number, compared with the nonglucocorticoid-treated group. Ucn1 mRNA was expressed in lamina propria plasma cells, and both corticotropin releasing factor(1) and corticotropin-releasing factor(2(a)) mRNAs were also partially coexpressed in these cells and macrophages. The present study showed that Ucn1-positive cells were correlated with the severity of inflammation in colonic mucosa with UC, and glucocorticoid treatment decreased these cells. Ucn1 therefore may act as a possible local immune-inflammatory mediator in UC. PMID- 15531482 TI - Prognostic factors associated with the survival of patients developing loco regional recurrences of differentiated thyroid carcinomas. AB - To estimate survival of patients with loco-regional recurrences (LRRs) of differentiated thyroid carcinomas (DTCs) and to identify factors associated with survival after LRRs, we analyzed retrospective data of the 172 patients treated and followed up in our institution from 1958 to 2000 who had developed LRRs (6% of DTC patients). Ultrasound, when used, picked up 95% of the recurrences. Survival was estimated with the method of Kaplan-Meier, and associated prognostic features were studied in univariate and multivariate Cox model-based analyses. Cumulated survival rates 10 yr after LRRs were 49.1, 89.3, and 32.1% for all patients, patients aged less than 45 yr, and older patients, respectively. Multivariate analysis identified three features related to initial tumor (age >/= 45 yr, follicular histology, presence of thyroid capsular effraction), the absence of radioiodine ablation of thyroid remnants after initial surgery (10% of patients did not receive radioiodine), the presence of distant metastases before LRR diagnosis, and two features related to the LRRs (no radioiodine uptake and thyroid bed location) as significantly associated with a reduced survival. Our results underline the seriousness of LRRs of DTCs and could be used to identify patients who should benefit from a closer follow-up and especially reactive therapeutic intervention. PMID- 15531483 TI - Decreased quality of life in patients with acromegaly despite long-term cure of growth hormone excess. AB - The long-term impact of acromegaly on subjective well-being after treatment of GH excess is unclear. Therefore, we evaluated quality of life by validated questionnaires in a cross-sectional study of 118 successfully treated acromegalic patients. The initial treatment was transsphenoidal surgery in most patients (92%), if necessary followed by radiotherapy or octreotide. All patients were in remission at the time of assessment (GH, <1.9 mug/liter; normal IGF-I for age). General perceived well-being was reduced compared with controls for all subscales (P < 0.001) as measured by the Nottingham Health Profile and the Short Form-36. Acromegalic patients also had lower scores on fatigue (Multidimensional Fatigue Index) and anxiety and depression (Hospital Anxiety and Depression Scale). Radiotherapy was associated with decreased quality of life in all subscales except for the Hospital Anxiety and Depression Scale, and worsened quality of life significantly, according to the fatigue scores. Somatostatin analog treatment was not associated with improved quality of life. Independent predictors of quality of life were age (physical subscales and Nottingham Health Profile), disease duration (social isolation and personal relations), and radiotherapy (physical and fatigue subscales). In conclusion, patients cured after treatment for acromegaly have a persistently decreased quality of life despite long-term biochemical cure of GH excess. Radiotherapy especially is associated with a reduced quality of life. PMID- 15531484 TI - Plasma von Willebrand factor and the development of the metabolic syndrome in patients with hypertension. AB - Although the metabolic syndrome is associated with endothelial damage/dysfunction, the effect of risk factors and their relationship with the development of this condition are unclear. We hypothesized that plasma von Willebrand factor (vWf, marking endothelial damage/dysfunction) increases with the number of components of metabolic syndrome and that increased levels precede its development. To test this, fasting vWf, glucose and lipids were measured in 161 patients (mean age 63 +/- 7 yr, 85% males) with hypertension. Using World Health Organization (WHO) criteria, 32 (19.9%), and using National Cholesterol Education Program (NCEP) criteria, 70 (43.5%) had metabolic syndrome. Plasma vWf was higher in these patients regardless of defining criteria and increased with the number of the components of metabolic syndrome (both P < 0.001). After 4 yr, patients who did not have metabolic syndrome at baseline were reassessed for the development of this condition. Of the 129 patients who did not meet the WHO criteria at baseline, 38 (29.5%) subsequently developed the condition, whereas 36 of the 91 (39.6%) who did not meet the NCEP criteria at baseline subsequently developed metabolic syndrome. Baseline vWf levels did not predict development of metabolic syndrome, regardless of criteria (P = 0.071 for WHO and P = 0.639 for NCEP). Our data suggest more severe endothelial damage/dysfunction with cumulative metabolic syndrome-related risk factors. The failure of plasma vWf to predict the development of metabolic syndrome suggests that endothelial damage/dysfunction is a consequence, not a cause, of these risk factors. PMID- 15531485 TI - Intensive insulin treatment in critically ill trauma patients normalizes glucose by reducing endogenous glucose production. AB - Critical illness is associated with insulin resistance and hyperglycemia. Intensive insulin treatment to normalize blood glucose during feeding has been shown to improve morbidity and mortality in patients in intensive care. The mechanisms behind the glucose-controlling effects of insulin in stress are not well understood. Six previously healthy, severely traumatized patients (injury severity score > 15) were studied early (24-48 h) after trauma. Endogenous glucose production (EGP) and whole-body glucose disposal (WGD) were measured (6,6 (2)H(2)-glucose) at basal, during total parenteral nutrition (TPN), and during TPN plus insulin to normalize blood glucose (TPN+I). Six matched volunteers served as controls. At basal and TPN, concentrations of glucose and insulin were higher in patients (P < 0.05). During TPN+I, insulin concentrations were 30-fold higher in patients. At basal, WGD and EGP were 30% higher in patients (P < 0.05). During TPN, EGP decreased in both groups but less in patients, resulting in 110% higher EGP than controls (P < 0.05). Normoglycemia coincided with reduced EGP, resulting in similar rates in both groups. WGD did not change during TPN or TPN+I and was not different between the groups. In conclusion, in healthy subjects, euglycemia is maintained during TPN at physiological insulin concentrations by a reduction of EGP, whereas WGD is maintained at basal levels. In traumatized patients, hyperglycemia is due to increased EGP. In contrast to controls, normalization of glucose concentration during TPN needs high insulin infusion rates and is accounted for by a reduction in EGP, whereas WGD is not increased. PMID- 15531486 TI - Vitamin D2 is much less effective than vitamin D3 in humans. AB - Vitamins D(2) and D(3) are generally considered to be equivalent in humans. Nevertheless, physicians commonly report equivocal responses to seemingly large doses of the only high-dose calciferol (vitamin D(2)) available in the U.S. market. The relative potencies of vitamins D(2) and D(3) were evaluated by administering single doses of 50,000 IU of the respective calciferols to 20 healthy male volunteers, following the time course of serum vitamin D and 25 hydroxyvitamin D (25OHD) over a period of 28 d and measuring the area under the curve of the rise in 25OHD above baseline. The two calciferols produced similar rises in serum concentration of the administered vitamin, indicating equivalent absorption. Both produced similar initial rises in serum 25OHD over the first 3 d, but 25OHD continued to rise in the D(3)-treated subjects, peaking at 14 d, whereas serum 25OHD fell rapidly in the D(2)-treated subjects and was not different from baseline at 14 d. Area under the curve (AUC) to d 28 was 60.2 ng.d/ml (150.5 nmol.d/liter) for vitamin D(2) and 204.7 (511.8) for vitamin D(3) (P < 0.002). Calculated AUC(infinity) indicated an even greater differential, with the relative potencies for D(3):D(2) being 9.5:1. Vitamin D(2) potency is less than one third that of vitamin D(3). Physicians resorting to use of vitamin D(2) should be aware of its markedly lower potency and shorter duration of action relative to vitamin D(3). PMID- 15531487 TI - Changes in plasma ghrelin concentration immediately after gastrectomy in patients with early gastric cancer. AB - Although the majority of circulating ghrelin originates from the stomach, no prospective study of the proportion of ghrelin derived from the stomach has been reported. Patients with early gastric cancer who underwent gastric resection were divided into three groups according to the extent and site of gastric resection: subtotal gastrectomy group (n = 24), proximal gastrectomy group (n = 4), and total gastrectomy group (n = 12). Patients with advanced gastric cancer who underwent gastrojejunostomy without gastrectomy served as the bypass group (n = 5). Blood samples were collected from all patients preoperatively, at 1 h after gastric resection or gastrojejunostomy, and on postoperative d 1, 3, and 7. The plasma ghrelin level was determined in all samples and expressed as a percentage of the preoperative level. In the bypass group, no significant drop in the ghrelin level was observed at 1 h after gastrojejunostomy, and the ghrelin level remained stable through postoperative d 7. In the subtotal gastrectomy group, the ghrelin concentration reached a nadir of 38.8 +/- 12.9% of preoperative levels at 1 h after gastric resection and then gradually increased to 88.1 +/- 13.2% by postoperative d 7. In the proximal gastrectomy group, the nadir ghrelin level was 24.5 +/- 15.4% at 1 h after gastric resection and was followed by a gradual recovery. However, the recovery rate was slower than that in the subtotal gastrectomy group, with the ghrelin level reaching only 47.6 +/- 18.8% by postoperative d 7 (P < 0.05). In the total gastrectomy group, the nadir ghrelin level was 28.6 +/- 11.1% at 1 h after gastric resection and remained at 30.0 +/- 13.2% until postoperative d 7. These results suggest that compensatory ghrelin production can occur in the remnant stomach after the surgical removal of part of the stomach and that the proximal fundus is more important than the distal antrum and body in terms of the capacity for ghrelin production. The principal site of ghrelin production is clearly the stomach, which contributes 70% of the circulating ghrelin concentration. PMID- 15531488 TI - Effect of recombinant human growth hormone (GH) replacement on the hypothalamic pituitary-adrenal axis in adult GH-deficient patients. AB - The aim of the study was to evaluate the hypothalamus-pituitary-adrenal (HPA) axis in patients (nine males, three females; mean age +/- sem 51 +/- 2 yr) with adult-onset GH deficiency (GHD) due to surgically treated pituitary tumors with preserved HPA function and without evidence of tumor recurrence before and during recombinant human (rh) GH replacement therapy (duration 31 +/- 6 months). HPA function was assessed by urinary free cortisol and morning serum cortisol levels as well as cortisol responses to 1 mug ACTH test (n = 7 patients) or insulin tolerance test (n = 5 patients) before and during rhGH therapy, the cut-off for the diagnosis of hypoadrenalism being a cortisol peak less than 18 microg/dl (<500 nmol/liter) after stimulatory tests. Serum cortisol and urinary free cortisol levels were significantly lower on therapy than before [7.6 +/- 0.8 vs. 11.5 +/- 0.9 microg/dl (208 +/- 22 vs. 317 +/- 24 nmol/liter), P < 0.01, and 19.6 +/- 2.5 vs. 32.2 +/- 3.2 microg per 24 h (54 +/- 7 vs. 89 +/- 9 nmol per 24 h), P < 0.05, respectively], whereas no change in cortisol-binding globulin levels was observed. Cortisol peak after either ACTH test or insulin tolerance test was lower on rhGH therapy than before [15.9 +/- 1.5 vs. 20.2 +/- 1.1 microg/dl (437 +/- 43 vs. 557 +/- 31), P = 0.01, and 13.1 +/- 2.6 vs. 20.4 +/- 1.4 microg/dl (362 +/- 71 vs. 564 +/- 37 nmol/liter), P = 0.03, respectively]. Accordingly, central hypoadrenalism was detected in nine of 11 patients. In conclusion, low GH and IGF-I levels, likely enhancing the conversion of cortisone to cortisol, may mask a condition of central hypoadrenalism. Therefore, the reassessment of HPA function in GHD patients during rhGH therapy is mandatory. PMID- 15531489 TI - Endocrine and metabolic effects of physiologic r-metHuLeptin administration during acute caloric deprivation in normal-weight women. AB - Leptin is a nutritionally regulated hormone that may modulate neuroendocrine function during caloric deficit. We hypothesized that administration of low-dose leptin would prevent changes in neuroendocrine function resulting from short-term caloric restriction. We administered physiologic doses of r-metHuLeptin [(0.05 mg/kg sc daily or identical placebo in divided doses (0800, 1400, 2000, and 0200 h)] to 17 healthy, normal-weight, reproductive-aged women during a 4-d fast. Leptin levels were lower in the placebo-treated group during fasting (3.3 +/- 0.2 vs. 9.6 +/- 1.0 ng/ml, P < 0.001, placebo vs. leptin-treated at end of study). Fat mass decreased more in the leptin than the placebo-treated group (-0.6 +/- 0.1 vs. -0.2 +/- 0.1 kg, P = 0.03). Both overnight LH area (38.9 +/- 21.5 vs. 1.2 +/- 11.1 microIU/ml.min, P = 0.05) and LH peak width increased (15.8 +/- 7.1 vs. 2.3 +/- 6.7 min, P = 0.06) and LH pulsatility decreased (-2.0 +/- 0.9 vs. 1.0 +/- 0.8 peaks/12 h, P = 0.03) more in the leptin vs. placebo group. LH pulse regularity was higher in the leptin-treated group (P = 0.02). Twenty-four-hour mean TSH decreased more in the placebo than the leptin-treated group, respectively (-1.06 +/- 0.27 vs. -0.32 +/- 0.18 microIU/ml, P = 0.03). No differences in 24-h mean GH, cortisol, IGF binding protein-1, and IGF-I were observed between the groups. Hunger was inversely related to leptin levels in the subjects randomized to leptin (r = -0.76, P = 0.03) but not placebo (r = -0.18, P = 0.70) at the end of the study. Diminished hunger was seen among subjects achieving the highest leptin levels. Our data provide new evidence of the important role of physiologic leptin regulation in the neuroendocrine response to acute caloric deprivation. PMID- 15531490 TI - gamma-Glutamyltransferase, obesity, and the risk of type 2 diabetes: observational cohort study among 20,158 middle-aged men and women. AB - Serum gamma-glutamyltransferase (GGT) concentration within its normal range has emerged as an important predictor in the pathogenesis of diabetes. We studied serum GGT as a predictor of type 2 diabetes incidence and a possible interaction between obesity and GGT on the development of type 2 diabetes in men and women. A prospective cohort study of 20,158 Finnish men and women aged 25-64 yr who participated in cardiovascular risk-factor surveys carried out in four areas during 10 yr. The average follow-up time was 12.7 yr, and there were 388 incident diabetes cases. Serum GGT cut points were at the 25th, 50th, 75th, and 90th percentiles. Initiation of new diabetes medication defined incidence cases. After adjustment for known risk factors of type 2 diabetes, relative risks for incident diabetes across GGT categories were 1.0, 1.2, 2.3, 3.1, and 3.9 among men and 1.0, 0.8, 1.7, 3.5, and 6.4 among women (P for trend < 0.01, respectively). Body mass index appeared to be more strongly associated with type 2 diabetes in both men and women over age 50 yr with GGT median or greater, compared with subjects with GGT less than median. In conclusion, in women as well as men, serum GGT level within its normal range predicted type 2 diabetes and may modify the well known association between body mass index and type 2 diabetes. PMID- 15531491 TI - Randomized controlled clinical trial of surgery versus no surgery in patients with mild asymptomatic primary hyperparathyroidism. AB - Parathyroidectomy is the definitive therapy for patients with symptomatic primary hyperparathyroidism. However, the role of surgery in mild asymptomatic primary hyperparathyroidism remains controversial. Accordingly, we conducted a prospective, randomized, controlled clinical trial of parathyroidectomy to determine the benefits of surgery vs. adverse effects of no surgery. Fifty-three patients were randomly assigned to either parathyroidectomy (n = 25) or regular follow-up (n = 28). Bone mineral density (BMD), biochemical indices of the disease, quality of life, and psychological function were measured at 6- or 12 month intervals for at least 24 months. Twenty-three of the 25 patients randomized to parathyroidectomy had surgery within the specified time of the protocol and three of the 28 patients randomized to regular follow-up had parathyroidectomy during follow-up. After parathyroidectomy, there was an increase in BMD of the spine (1.2%/yr, P < 0.001), femoral neck (0.4%/yr, P = 0.031), total hip (0.3%/yr, P = 0.07), and forearm (0.4%/yr, P < 0.001) and an expected fall in serum total and ionized calcium, serum PTH, and urine calcium (P < 0.001 for all). In contrast, patients followed up without surgery lost BMD at the femoral neck (-0.4%/yr, P = 0.117) and total hip (-0.6%/yr, P = 0.007) but gained at the spine (0.5%/yr; P = ns) and forearm (0.2%/yr, P = 0.047), with no significant changes in biochemical indices of disease. Consequently, a significant effect of parathyroidectomy on BMD was evident only at the femoral neck (a group difference of 0.8%/yr; P = 0.01) and total hip (a group difference of 1.0%/yr; P = 0.001) but not at the spine (a group difference of 0.6%/yr) or forearm (a group difference of 0.2%/yr). Quality-of-life scores as measured by a 36-item short-form health survey showed significant declines in five of the nine domains (social functioning, physical problem, emotional problem, energy, and health perception) in patients followed up without surgery but in only one of the nine domains (physical function) in the patients who had parathyroidectomy. Consequently, a modest measurable benefit of parathyroidectomy was evident in social and emotional role function (P = 0.007 and 0.012, respectively). Psychological function as assessed by the symptom checklist revised did not change significantly in either group, except for a significant decline in anxiety (P = 0.003) and phobia (P = 0.024) in patients who had surgery in comparison with those who did not. We conclude that it is feasible to conduct a randomized, controlled clinical trial of parathyroidectomy in patients with mild asymptomatic primary hyperparathyroidism, and measurable benefits of surgery on BMD, quality of life, and psychological function can be demonstrated. However, the small but significant benefits of parathyroidectomy must be weighed against the risks of surgery in these otherwise healthy individuals. PMID- 15531492 TI - Increased plasma S100A12 (EN-RAGE) levels in patients with type 2 diabetes. AB - S100A12, also called EN-RAGE (extracellular newly identified receptor for advanced glycation end products binding protein) or calcium-binding protein in amniotic fluid-1, is a ligand for RAGE. It has been shown that S100A12 induces adhesion molecules such as vascular cell adhesion molecule-1 and intercellular adhesion molecule-1 in the vascular endothelial cell and mediates migration and activation of monocytes/macrophages through RAGE binding and that infusion of lipopolysaccharide into mice causes time-dependent increase of S100A12 in the plasma. Therefore, circulating S100A12 protein may be involved in chronic inflammation in the atherosclerotic lesion. In this study, we developed an ELISA system that uses specific monoclonal antibodies against recombinant human S100A12 to measure plasma S100A12 levels in patients with diabetes. On using our S100A12 ELISA system, the coefficients of variation of intra- and interassay were less than 4 and 9%, respectively. The analytical lower detection limit was 0.2 ng/ml. When plasma S100A12 levels were measured by this system, the concentrations were more than twice as high in the patients with diabetes, compared with those without. Using univariate analysis in all subjects, plasma S100A12 concentrations correlated with hemoglobin A1c, fasting glucose, high-sensitivity C-reactive protein and white blood cell count. Stepwise multiple regression analyses, however, revealed that only white blood cell count and hemoglobin A1c remained significant independent determinants of plasma S100A12 concentration. These results suggest that plasma S100A12 protein levels are regulated by factors related to subclinical inflammation and glucose control in patients with type 2 diabetes. PMID- 15531493 TI - Intramuscular testosterone undecanoate: pharmacokinetic aspects of a novel testosterone formulation during long-term treatment of men with hypogonadism. AB - In an open-label, randomized, prospective trial, we investigated pharmacokinetics and several efficacy and safety parameters of a novel, long-acting testosterone (T) undecanoate (TU) formulation in 40 hypogonadal men (serum testosterone concentrations < 5 nmol/liter). For the first 30 wk (comparative study), the patients were randomly assigned to receive either 10 x 250 mg T enanthate (TE) im every 3 wk (n = 20) or 3 x 1000 mg TU im every 6 wk (loading dose) followed by 1 x 1000 mg after an additional 9 wk (n = 20). In a follow-up study, observation continued in those patients who completed the comparative part and opted for TU treatment (8 x 1000 mg TU every 12 wk in former TU patients and 2 x 1000 mg TU every 8 wk plus 6 x 1000 mg every 12 wk in former TE patients) for an additional 20-21 months. Here we report only the pharmacokinetic aspects of the new TU formulation for the first approximately 2.5 yr of treatment. At baseline, serum T concentrations did not significantly differ between the two study groups. In the TE group, mean trough levels of serum T were always less than 10 nmol/liter before the next injection, whereas in the TU group, mean trough levels of serum T were 14.1 +/- 4.5 nmol/liter after the first two doses (6-wk intervals) and 16.3 +/- 5.7 nmol/liter after the 9-wk interval at wk 30. The mean serum levels of dihydrotestosterone and estradiol also increased in parallel to the serum T pattern and remained within the normal range. In the follow-up study, the former TU patients (n = 20) received eight TU injections at 12-wk intervals, and the TE patients (n = 16) switched to TU and initially received two TU injections at 8-wk intervals (loading) and continued with six TU injections at 12-wk intervals (maintenance). This regimen resulted in stable mean serum trough levels of T (ranging from 14.9 +/- 5.2 to 16.5 +/- 8.0 nmol/liter) and estradiol (ranging from 98.5 +/- 45.2 to 80.4 +/- 14.4 pmol/liter). The present study has shown that 1000 mg TU injected into male patients with hypogonadism at 12-wk intervals is well tolerated and leads to T levels within normal ranges, using four instead of 17 or more TE injections per year. An initial loading dose of either 3 x 1000 mg TU every 6 wk at the beginning of hormone substitution or 2 x 1000 mg TU every 8 wk after switching from the short-acting TE to TU were found to be a adequate dosing regimens for starting of treatment with the long-acting TU preparation. PMID- 15531494 TI - Changes in height, weight, and body mass index in children with craniopharyngioma after three years of growth hormone therapy: analysis of KIGS (Pfizer International Growth Database). AB - Extreme degrees of obesity may occur in association with hypothalamic tumors, usually after surgical intervention. This phenomenon has been reported to occur in as many as 25-75% of children undergoing extensive surgical extirpation of craniopharyngiomas (Cranio). Because less is known about the auxology of children with Cranio with milder alterations in growth, we undertook a 3-yr longitudinal analysis, using the KIGS database (Pfizer International Growth Database), to study their growth patterns and evolution of weight. We compared the effect of GH therapy on height, weight, and body mass index (BMI) in 199 prepubertal children with diagnosed Cranio treated by surgery and/or radiotherapy to two other groups of children with other causes of organic GH deficiency (OGHD): one with postsurgical and/or postirradiated OGHD (OGHD + S/I; n = 92) and the other with OGHD not due to Cranio and not having undergone either surgery or irradiation (OGHD - S/I; n = 85). At the start of GH therapy, 1) mean chronological (P < 0.0001) and bone (P = 0.0002) ages were youngest in OGHD - S/I and oldest in OGHD + S/I; 2) the mean height sd score (SDS) was lowest in OGHD - S/I and comparably higher in the other two groups (P < 0.0001); 3) mean weight and BMI SDS were greatest in Cranio and least in OGHD - S/I (both P < 0.0001); and 4) the mean initial GH dose prescribed was highest in OGHD - S/I and comparable in the other two groups (P < 0.0001). After 3 yr of GH therapy, 1) mean bone age remained youngest in OGHD - S/I and oldest in OGHD + S/I (P < 0.0001); 2) mean height SDS was highest in Cranio and comparably lower in the other two groups (P = 0.0159); 3) mean weight and BMI SDS remained greatest in Cranio and least in OGHD - S/I (P < 0.0001 and P = 0.0003, respectively); and 4) the mean GH dose remained highest in the OGHD - S/I group and least in the Cranio group (P = 0.0082). There were statistically significant increases within each group between the start of treatment and after 3 yr of GH therapy in height and weight, but not in BMI SDS. Lastly, after 3 yr of GH treatment, children in the Cranio group continued to have disproportionately heavier weight and higher BMI (with the greatest values in those with lower stimulated peak GH concentrations) compared with members of the other two groups, with no salutary effect of GH treatment on weight SDS and a mild improvement in BMI SDS. After S/I treatment, children with Cranio are disproportionately prone to varying degrees of weight gain compared with children with other forms of OGHD. In the present cohort of prepubertal children with Cranio, GH therapy induced excellent linear growth, but failed to have an ameliorative effect on weight gain and had only a slight beneficial effect on BMI gain. Because affected children may have resultant significant long-term medical morbidity and diminished quality of life, it is critical that the mechanism of this phenomenon be determined to devise helpful preventive or therapeutic interventions. PMID- 15531495 TI - The effects of vitamin A deficiency and vitamin A supplementation on thyroid function in goitrous children. AB - In developing countries, children are at high risk for both the iodine deficiency disorders (IDD) and vitamin A deficiency (VAD). The study aim was to determine the effects of VAD and vitamin A (VA) supplementation on thyroid function in an area of endemic goiter. In a double-blind, randomized, 10-month trial, Moroccan children with IDD and VAD (n = 138) were given iodized salt and either VA (200,000 IU) or placebo at 0 and 5 months. At 0, 5, and 10 months, measurements of VA status and thyroid function were made. At baseline, increasing VAD severity was a predictor of greater thyroid volume and higher concentrations of TSH and thyroglobulin (P < 0.001). In children with VAD, the odds ratio for goiter was 6.51 (95% confidence interval, 2.94, 14.41). VAD severity was also a strong predictor of higher concentrations of total T(4) (P < 0.001); the odds ratio for hypothyroidism in VAD was 0.06 (95% confidence interval, 0.03, 0.14). During the intervention, mean thyroglobulin, median TSH, and the goiter rate significantly decreased in the VA-treated group compared with those in the placebo group (P < 0.01). The findings indicate that VAD in severely IDD-affected children increases TSH stimulation and thyroid size and reduces the risk for hypothyroidism. This effect could be due to decreased VA-mediated suppression of the pituitary TSHbeta gene. In IDD- and VAD-affected children receiving iodized salt, concurrent VA supplementation improves iodine efficacy. PMID- 15531496 TI - Serum adiponectin is reduced in acromegaly and normalized after correction of growth hormone excess. AB - Adiponectin, an adipocyte-derived hormone, possesses insulin-sensitizing, antiinflammatory, and antiatherogenic properties. We hypothesized that hypoadiponectinemia was present in acromegaly, as in other conditions with increased insulin resistance and cardiovascular risk. Using an in-house RIA, serum adiponectin was determined in 35 patients with active acromegaly and 35 age , sex-, and body mass index-matched healthy controls. Twenty-five patients were restudied after GH-lowering therapies. Serum adiponectin was significantly reduced in the acromegalic patients (4.3 +/- 1.8 vs. 6.7 +/- 1.8 microg/ml in controls; P < 0.001), but was increased after treatment with Sandostatin LAR, a long-acting somatostatin analog (5.8 +/- 2.6 vs. 3.8 +/- 1.6 microg/ml pretreatment; P < 0.001; n = 15) or transsphenoidal surgery (6.5 +/- 2.7 vs. 3.9 +/- 1.5 microg/ml preoperation; P < 0.01; n = 10). Fasting insulin was an independent determinant of serum adiponectin levels (P < 0.01) in control subjects, contributing to 11.7% of the variance in circulating adiponectin. In cultured 3T3-L1 adipocytes, adiponectin mRNA levels were decreased by insulin (1.5 microm; P < 0.005) or IGF-I (1 microg/ml; P < 0.05), but not by GH (1 microm) or somatostatin (1 microm). In conclusion, hypoadiponectinemia is present in active acromegaly, probably secondary to the inhibitory effect of high circulating insulin levels. Hypoadiponectinemia, reversible with GH-lowering therapies, may contribute to the increased insulin resistance and cardiovascular risk in patients with acromegaly. PMID- 15531497 TI - Evidence for an association between metabolic cardiovascular syndrome and coronary and aortic calcification among women with polycystic ovary syndrome. AB - Women with polycystic ovary syndrome (PCOS) exhibit an adverse cardiovascular risk profile, characteristic of the metabolic cardiovascular syndrome (MCS). The aim of this study was to determine the prevalence of coronary artery (CAC) and aortic (AC) calcification among middle-aged PCOS cases and controls and to explore the relationship among calcification, MCS, and other cardiovascular risk factors assessed 9 yr earlier. This was a prospective study of 61 PCOS cases and 85 similarly aged controls screened in 1993-1994 for risk factors and reevaluated in 2001-2002. The main outcome measures were CAC and AC, measured by electron beam tomography. Women with PCOS had a higher prevalence of CAC (45.9% vs. 30.6%) and AC (68.9% vs. 55.3%) than controls. After adjustment for age and body mass index, PCOS was a significant predictor of CAC (odds ratio = 2.31; P = 0.049). PCOS subjects were also 4.4 times more likely to meet the criteria for MCS than controls. High-density lipoprotein cholesterol and insulin appeared to mediate the PCOS influence on CAC. Interestingly, total testosterone was an independent risk factor for AC in all subjects after controlling for PCOS, age, and body mass index (P = 0.034). We conclude that women with PCOS are at increased risk of MCS and demonstrate increased CAC and AC compared with controls. Components of MCS mediate the association between PCOS and CAC, independently of obesity. PMID- 15531498 TI - Frequent occurrence of hypogonadotropic hypogonadism in type 2 diabetes. AB - Type 2 diabetes is associated with lower total testosterone (T) levels in cross sectional studies. However, it is not known whether the defect is primary or secondary. We investigated the prevalence of hypogonadism in type 2 diabetes by measuring serum total T, free T (FT), SHBG, LH, FSH, and prolactin (PRL) in 103 type 2 diabetes patients. FT was measured by equilibrium dialysis. FT was also calculated by using T and SHBG (cFT). Hypogonadism was defined as low FT or cFT. The mean age was 54.7 +/- 1.1 yr, mean body mass index (BMI) was 33.4 +/- 0.8 kg/m(2), and mean HbA1c was 8.4 +/- 0.2%. The mean T was 12.19 +/- 0.50 nmol/liter (351.7 +/- 14.4 ng/dl), SHBG was 27.89 +/- 1.65 nmol/liter, and FT was 0.250 +/- 0.014 nmol/liter. Thirty-three percent of patients were hypogonadal. LH and FSH levels were significantly lower in the hypogonadal group compared with patients with normal FT levels (3.15 +/- 0.26 vs. 3.91 +/- 0.24 mIU/ml for LH and 4.25 +/- 0.45 vs. 5.53 +/- 0.40 mIU/ml for FSH; P < 0.05). There was a significant inverse correlation of BMI with FT (r = -0.382; P < 0.01) and T (r = 0.327; P < 0.01). SHBG correlated inversely with BMI (r = -0.267; P < 0.05) but positively with age (r = 0.538; P < 0.001) and T (r = 0.574; P < 0.001). FT correlated strongly with cFT (r = 0.919; P < 0.001) but not with SHBG. LH levels correlated positively with FT (r = 0.287; P < 0.05). We conclude that hypogonadotropic hypogonadism occurs commonly in type 2 diabetes. PMID- 15531499 TI - beta-Cell function and insulin sensitivity in early adolescence: association with body fatness and family history of type 2 diabetes mellitus. AB - The prevalence of type 2 diabetes mellitus (T2DM) among adolescents has increased 5- to 10-fold over the past decade. T2DM results from pancreatic beta-cell dysfunction and insulin resistance. Using rapid iv glucose tolerance testing, we examined beta-cell function and insulin resistance in 72 predominantly Latino eighth grade students (41 males and 31 females; mean +/- sem age, 13.6 +/- 0.1 yr). Thirty-six percent of the children had body mass indexes above the 85th percentile for age and gender, and 50% had a first- or second-degree relative with T2DM. Overweight children were five times more likely to be in the highest quartile for insulin resistance. Children with a family history of T2DM were five times more likely to be in the lowest quartile for insulin secretory capacity, 4.5 times more likely to be in the lowest quartile for glucose disposal, and three times more likely to be in the lowest quartile for insulin resistance. These findings are consistent with a model for the physiology of T2DM in which a familial beta-cell dysfunction is unmasked by increasing insulin resistance secondary to overweight in this predominantly Latino population. PMID- 15531500 TI - Serum parathyroid hormone is associated with increased mortality independent of 25-hydroxy vitamin d status, bone mass, and renal function in the frail and very old: a cohort study. AB - Very frail older people constitute an increasing proportion of the aging population and often have vitamin D deficiency and impaired renal function. Primary hyperparathyroidism has been associated with increased mortality, but it is unclear whether secondary hyperparathyroidism is associated with increased mortality independent of renal function and vitamin D status. This study aimed to examine the effect of vitamin D deficiency and secondary hyperparathyroidism on mortality in frail older people after accounting for renal function and general measures of health. We evaluated 842 subjects (182 men with a mean age of 81.9 yr and 660 women with a mean age of 86.2 yr) living in residential aged care facilities in Sydney, Australia in a prospective, cohort study. Over a mean duration of follow-up of 31 months, 345 subjects died. Baseline serum 25-hydroxy vitamin D, serum PTH, and bone ultrasound attenuation were significantly associated with mortality in univariate and multivariate analyses (for PTH, a hazard ratio of 1.39 for time to death) after correcting for age and gender. In multivariate analyses that corrected for health status, nutritional status, and renal function, PTH remained a significant predictor of mortality but not 25 hydroxy vitamin D or bone ultrasound attenuation. Serum PTH appears to be associated with increased mortality in the frail elderly independent of vitamin D status, renal function, bone mass, and measures of general health. The mechanism of this effect requires further investigation. PMID- 15531501 TI - Plasma levels of the endocannabinoid anandamide in women--a potential role in pregnancy maintenance and labor? AB - Although exposure to exocannabinoids (e.g. marijuana) is associated with adverse pregnancy outcome, little is known about the biochemistry, physiology, and consequences of endocannabinoids in human pregnancy. In these studies, we measured the levels of the endocannabinoid anandamide (N arachidonoylethanolamine, AEA) by HPLC-mass spectrometry in 77 pregnant and 25 nonpregnant women. The mean +/- sem plasma AEA levels in the first, second, and third trimesters were 0.89 +/- 0.14, 0.44 +/- 0.12, and 0.42 +/- 0.11 nm, respectively. The levels in the first trimester were significantly higher than those in either the second or third trimester. During labor, AEA levels were 3.7 times nonlaboring term levels (2.5 +/- 0.22 vs. 0.68 +/- 0.09 nm, P < 0.0001). During the menstrual cycle, levels in the follicular phase were significantly higher than those in the luteal phase (1.68 +/- 0.16 vs. 0.87 +/- 0.09 nm, P < 0.005). Postmenopausal and luteal-phase levels were similar to those in the first trimester. These findings suggest that successful pregnancy implantation and progression requires low levels of AEA. At term, AEA levels dramatically increase during labor and are affected by the duration of labor, suggesting a role for AEA in normal labor. PMID- 15531502 TI - Spontaneous growth hormone (GH) secretion is not directly affected by ghrelin in either short normal prepubertal children or children with GH neurosecretory dysfunction. AB - Ghrelin, a specific endogenous ligand for the GH secretagogue receptor, stimulates GH secretion in humans when given in pharmacological amounts. Under physiological conditions, however, it is controversial whether ghrelin affects GH secretion and vice versa. No studies have reported on the relationship between daily ghrelin and GH secretion in children. Therefore, plasma ghrelin and GH concentrations over a 24-h period were studied in 10 prepubertal short normal children (five females and five males) to determine the potential relationship between the secretion of these two hormones. Furthermore, five prepubertal patients (two females and three males) with GH neurosecretory dysfunction (GHNSD) were studied in the same way to assess potential alterations in ghrelin secretion in a condition associated with distinct GH changes. No gender difference in ghrelin spontaneous secretion was detected in either short normal children or GHNSD patients, and in both male and female subjects, ghrelin was secreted in a pulsatile and circadian fashion, with a nocturnal surge. Twenty-four-hour secretion and daytime ghrelin secretion of short normal children were similar to those in GHNSD patients, whereas nighttime hormone secretion in the latter group was significantly greater than that in short normal children. The cross correlation of 24-h ghrelin and GH levels revealed significant positive and negative correlations, which were similar in the two groups examined. The positive one, with GH leading ghrelin, might reflect a somatostatin (SMS) mediated inhibitory effect on both GH and ghrelin secretion (low SMS levels are followed by high GH and ghrelin levels, and vice versa). The negative correlation, with ghrelin leading GH, might again reflect the positive effect of ghrelin on SMS, as shown in both animal and human studies. In conclusion, the results of the present study indicate that ghrelin secretion in prepubertal children is pulsatile and is not sexually dimorphic. Although the parallelism of ghrelin and GH dynamics hints at the potential relevance of endogenous ghrelin as a promoter of GH release, our data do not support this hypothesis. We suggest that the interactions of ghrelin and GH are the result of SMS action. SMS inhibits GH secretion not only by a direct effect on the pituitary and by inhibiting hypothalamic GHRH, but also through the suppression of ghrelin release. PMID- 15531503 TI - High levels of circulating CXC chemokine ligand 10 are associated with chronic autoimmune thyroiditis and hypothyroidism. AB - CXC chemokine ligand 10 (CXCL10), an interferon-gamma-inducible chemokine associated with Th1-mediated immune responses, has been proposed as a marker of inflammation in autoimmune diseases. We measured serum CXCL10 concentrations in 223 consecutive patients with newly diagnosed autoimmune thyroiditis (AT), 97 euthyroid controls, and 29 patients with nontoxic multinodular goiter and related this parameter to the clinical phenotype. The three groups were similar in gender distribution and age; among the AT patients, 24% had subclinical hypothyroidism. Serum CXCL10 level was significantly higher in AT patients (157 +/- 139 pg/ml) than in controls (79 +/- 38) or patients with multinodular goiter (90 +/- 32; P < 0.0001). Among patients with AT, CXCL10 levels were significantly higher in those with a hypoechoic ultrasonographic pattern and hypothyroidism. In a multiple linear regression model including age, thyroid volume, hypoechogenicity, hypervascularity, TSH, free T(4), and antithyroid peroxidase, only age (standardized coefficient = 0.39; P = 0.0001) and TSH (standardized coefficient = 0.41; P < 0.0002) were significantly related to serum CXCL10 levels. We conclude that circulating CXCL10 is increased in patients with AT and is associated with hypothyroidism. CXCL10 may be regarded as a marker of a more aggressive thyroiditis leading to thyroid destruction. PMID- 15531504 TI - Adiponectin levels in the first two years of life in a prospective cohort: relations with weight gain, leptin levels and insulin sensitivity. AB - Adiponectin, a novel adipocytokine with insulin sensitizing properties, is inversely related to obesity and insulin resistance in adults. We recently reported large variations in weight gain and insulin sensitivity during the first year in infants born small for gestational age (SGA) or appropriate for gestational age (AGA). We now determined whether adiponectin levels were related to postnatal growth and insulin sensitivity in a prospective cohort followed from birth to two years old (n = 85) (55 female/30 male, 65 SGA/20 AGA). Serum adiponectin levels at one year and two years were higher compared to reported levels in adults and older children, and decreased from one year (21.6 +/- 0.6 microg/ml) to two years (15.7 +/- 0.7 microg/ml) (p < 0.05). At two years adiponectin levels were lower in females (15.3 +/- 0.4 microg/ml) than males (16.4 +/- 0.6 microg/ml) (p < 0.05), but no gender difference was seen in leptin or insulin levels. No differences in adiponectin levels were seen between SGA and AGA infants at one or two years. However, in SGA infants changes in adiponectin between one to two years old were inversely related to weight gain (r = -0.310, p < 0.05). Changes in leptin levels between one to two years were positively related to weight gain in both SGA and AGA infants (r = 0.450 and r = 0.500 respectively, both p < 0.05). Adiponectin levels were unrelated to insulin levels at one or two years, nor to change in insulin levels between one to two years. In multiple regression analysis, adiponectin levels were related only to postnatal age; omitting age from the model, the determinants of higher adiponectin levels were male gender (p = 0.03), lower postnatal body weight (p < 0.001), and higher birth weight SD score (p = 0.004). In conclusion, fall in serum adiponectin levels during the first two years of life were related to increasing age and greater weight gain SGA infants, but were unrelated to insulin sensitivity. PMID- 15531505 TI - Glibenclamide treatment in permanent neonatal diabetes mellitus due to an activating mutation in Kir6.2. AB - Permanent neonatal diabetes mellitus (PNDM) is a rare form of diabetes characterized by insulin-requiring hyperglycemia that is diagnosed within the first months of life. Recently, activating mutations in the gene encoding the ATP sensitive potassium-channel subunit Kir6.2 were identified in 10 PNDM patients. Tolbutamide-stimulated insulin secretion, demonstrated in 3 of these patients suggested that some PNDM patients may respond to oral sulfonylurea treatment. In this report, we describe an infant with PNDM due to an arginine-to-histidine substitution at position 201 (R201H) of the gene encoding Kir6.2. After insulin pump therapy for six months, he was shifted to oral glybenclamide therapy at a daily dose of 0.8 mg/kg. Basal c-peptide level increased by two fold during glybenclamide treatment, but no further elevation was observed following intravenous glucose administration. Outpatient, continuous glucose monitoring while on a normal infant diet demonstrated a marked improvement in glycemic control. This study demonstrates the feasibility of oral sulfonylurea treatment in PNDM patients with Kir6.2 mutations even during infancy, and the superiority of this approach over insulin administration. PMID- 15531506 TI - Overexpression of follicle-stimulating hormone receptor activates oncogenic pathways in preneoplastic ovarian surface epithelial cells. AB - It has been previously demonstrated that human ovarian cancer cells express FSH receptor (FSHR). However, whether FSHR plays a role in ovarian cancer development is still ambiguous. To investigate the role of FSHR in tumor progression, we overexpressed the receptor in SV40 Tag immortalized ovarian surface epithelium (OSE) cell lines (IOSE-80PC, a postcrisis line, and IOSE-398), which are preneoplastic and nontumorigenic. We compared the expression levels of several selected oncogenes in nontransfected (80PC), vector-transfected (80PCV), FSHR transfected IOSE (80PCF) cells, and established ovarian cancer cell lines (OVCAR 3 and SKOV-3). Significantly increased protein levels of epithelial growth factor receptor, HER-2/neu, and c-Myc, but not K-Ras, were observed in FSHR overexpressing 80PCF cells when compared with 80PCV cells. Constitutive phosphorylation of ERK1/2 was augmented in 80PCF cells, whereas phosphorylation of the other MAPK including p38 and Jun N-terminal kinase was unchanged. Considerable constitutive phosphorylation of ERK1/2 was also observed in OVCAR-3 and SKOV-3 cell lines when compared with 80PCV. More importantly, 80PCF cells grew more rapidly than 80PC and 80PCV cells. In conclusion, we have demonstrated that FSHR was highly expressed in OVCAR-3 and 80PCF cells transfected with FSHR overexpression vector. The 80PCF cell line showed increased levels of epithelial growth factor receptor, HER-2/neu, and c-myc and constitutive activation of ERK1/2. The rate of proliferation of the 80PCF cells was increased, compared with control cell lines. These results suggest that the overexpression of FSHR may be associated with enhanced levels of potential oncogenic pathways and increased proliferation in preneoplastic ovarian surface epithelial cells. PMID- 15531507 TI - Nodal induces apoptosis and inhibits proliferation in human epithelial ovarian cancer cells via activin receptor-like kinase 7. AB - Human epithelial ovarian cancer is the most lethal female cancer. Hormones and growth factors, including the TGF-beta superfamily, have been suggested to play a role in ovarian tumorigenesis. The biological effects of TGF-beta superfamily are mediated by type I and type II serine/threonine kinase receptors and by intracellular Smad proteins. Recently, we have cloned four transcripts of human activin receptor-like kinase 7 (ALK7), a type I receptor for Nodal. In this study, we have investigated the role of Nodal and ALK7 in four ovarian cancer cell lines, OV2008, C13*, A2780-s, and A2780-cp. Overexpression of Nodal resulted in a significant decrease in the number of metabolically active cells. This effect was mimicked by a constitutively active ALK7 (ALK7-ca) but blocked by dominant negative mutants of ALK7, Smad2, or Smad3. Transient transfection of Nodal and ALK7-ca significantly decreased X-linked inhibitor of apoptosis protein (Xiap) expression, activated both caspase-3 and caspase-9, and increased apoptosis as determined by Hoechst nuclear staining and flow cytometry. In addition, Nodal and ALK7-ca also inhibited cell proliferation as measured by 5 bromo-2'-deoxyuridine (BrdU) assays. Interestingly, the effects of Nodal and ALK7 ca were more potent in chemosensitive A2780-s cells than in its chemoresistant counterpart, A2780-cp cells. These findings demonstrate that Nodal induces apoptosis and inhibits proliferation via ALK7 and Smad2/3 and that the effect of Nodal-ALK7 on apoptosis may be mediated in part by the down-regulation of Xiap and activation of caspase-9 and caspase-3. PMID- 15531508 TI - Pancreatic islets from type 2 diabetic patients have functional defects and increased apoptosis that are ameliorated by metformin. AB - Several properties of pancreatic beta-cells in type 2 diabetes (T2D) were studied by using islets isolated from T2D subjects. Moreover, because metformin has protective effects on nondiabetic beta-cells exposed to high glucose or free fatty acid levels, we investigated its direct action on T2D islet cells. Diabetic islets were characterized by reduced insulin content, decreased amount of mature insulin granules, impaired glucose-induced insulin secretion, reduced insulin mRNA expression, and increased apoptosis with enhanced caspase-3 and -8 activity. These alterations were associated with increased oxidative stress, as shown by higher nitrotyrosine concentrations, increased expression of protein kinase C beta2 and nicotinamide adenine dinucleotide phosphate reduced-oxidase, and changes in mRNA expression of manganese- superoxide dismutase, Cu/Zn-superoxide dismutase, catalase, and glutathione peroxidase. Twenty-four-hour incubation of T2D islets with metformin was associated with increased insulin content, increased number and density of mature insulin granules, improved glucose-induced insulin release, and increased insulin mRNA expression. Moreover, apoptosis was reduced, with concomitant decrease of caspase-3 and -8 activity. These changes were accompanied by reduction or normalization of several markers of oxidative stress. Thus, T2D islets have several functional and survival defects, which can be ameliorated by metformin; the beneficial effects of the drug are mediated, at least in part, by a reduction of oxidative stress. PMID- 15531509 TI - Age and secretagogue type jointly determine dynamic growth hormone responses to exogenous insulin-like growth factor-negative feedback in healthy men. AB - The primary cause of waning GH and IGF-I concentrations in healthy aging adults is not established. To test the postulate that age influences negative feedback by IGF-I in a secretagogue-specific fashion, 17 normal men (nine young and eight older) each completed eight randomly ordered injections of placebo or recombinant human (rh) IGF-I (20 microg/kg sc), followed by saline/rest, aerobic exercise, GHRH (1 microg/kg iv bolus), or GH-releasing peptide-2 (1 microg/kg iv bolus) stimulation. GH secretion was monitored by sampling blood every 10 min for 7 h, high-sensitivity immunochemiluminometric assay, and deconvolution analysis conditioned on prior pulse-onset times and biexponential kinetics. Analysis of covariance showed that age (P = 0.028), secretagogue (P < 0.001), and rhIGF-I (P < 0.005) individually determine pulsatile GH secretion and exhibit a strong 3 fold interaction (P < 10(-5)). Post hoc comparisons revealed that elderly subjects manifest less IGF-I inhibition of a maximal GHRH stimulus (P = 0.013 vs. young), blunted initial IGF-I suppression of fasting GH release (P = 0.038), and impaired IGF-I feedback on the regularity of GH secretion (P = 0.023). Age stratum did not influence peak IGF-I and nadir GH concentrations or rhIGF-I induced inhibition of GH secretion stimulated by exercise or GH-releasing peptide 2. In summary, experimental elevation of IGF-I concentrations unmasks reduced rhIGF-I-dependent feedback inhibition of fasting and GHRH-stimulated GH secretion in healthy older men, indicating that aging selectively modulates the autoinhibition process. PMID- 15531510 TI - Gonadotropin releasing hormone and transforming growth factor beta activate mitogen-activated protein kinase/extracellularly regulated kinase and differentially regulate fibronectin, type I collagen, and plasminogen activator inhibitor-1 expression in leiomyoma and myometrial smooth muscle cells. AB - GnRH analog (GnRHa) and TGF-beta act directly on leiomyoma/myometrial smooth muscle cells (LSMCs and MSMCs) regulating diverse activities resulting in leiomyoma growth and regression. Because GnRH and TGF-beta receptor signaling is in part mediated through the MAPK pathway, we determined whether the contribution of MAPK/ERK and transcriptional activation of c-fos and c-jun, result in differential regulation of type I collagen, fibronectin, and plasminogen activator inhibitor 1 (PAI-1) gene expression, whose products are known to influence extracellular matrix turnover, which is critical in leiomyoma growth and GnRHa-induced regression. We found that GnRHa and TGF-beta in a dose- and time-dependent manner increased the level of phosphorylated ERK1/2 (pERK1/2) in LSMCs and MSMCs. GnRHa and TGF-beta increased ERK1/2 nuclear accumulation resulting in differential regulation of c-fos and c-jun mRNA expression via downstream signaling from MAPK kinase (MEK)1/2, because pretreatment with U0126, a synthetic inhibitor of MEK1/2, abolished basal and GnRHa- and TGF-beta-induced pERK1/2 and the expression of c-fos and c-jun. LSMCs and MSMCs also express fibronectin, type I collagen, and PAI-1 mRNA, and GnRHa and TGF-beta altered their expression in a cell-specific manner through MEK1/2. We concluded that GnRHa and TGF-beta acting through a MAPK/ERK pathway and transcriptional activation of c-fos/c-jun results in differential regulation of specific genes whose products may in part influence the outcome of leiomyoma growth and regression. PMID- 15531511 TI - Stability of adrenocortical steroidogenesis over time in healthy women and women with polycystic ovary syndrome. AB - Adrenocortical secretion is up-regulated in women with polycystic ovary syndrome (PCOS), and absolute adrenal androgen (AA) excess is evident in approximately 25% of these patients. We hypothesized that AA biosynthesis is an inherited trait and that, as for other inherited traits, AA biosynthesis remains stable over time. To test this hypothesis, we prospectively studied 23 off-treatment PCOS patients and seven age- and body mass index-matched control women on two separate occasions 3 5 yr apart (45.0 +/- 19.0 months and 47.4 +/- 21.3 months, respectively; P > 0.05). All subjects underwent an acute adrenal stimulation using 0.25 mg ACTH-(1 24), and dehydroepiandrosterone (DHEA), androstenedione, and cortisol (F) were measured 0 and 60 min post ACTH; basal levels of total and free testosterone (T), SHBG, and DHEA sulfate (DHEA-S) were also assessed. Among PCOS patients, the mean DHEA-S levels during the repeat study were significantly lower when compared with the initial assessment (170 +/- 107 microg/dl vs. 134 +/- 79 microg/dl, respectively; P = 0.02). However, only patients with initial DHEA-S levels above the median (high DHEA-S) experienced a net decrease in the levels of this metabolite (252.5 +/- 99.2 microg/dl vs. 174.3 +/- 82.5 microg/dl; P = 0.001) over the time of the study; patients with initial DHEA-S levels in the lower half (low DHEA-S) did not experience a change in DHEA-S (94.6 +/- 28.9 microg/dl vs. 97.7 +/- 56.5 microg/dl; P = 0.85). In patients, the total T levels tended to be higher at the second study, although SHBG levels were also higher, resulting in unchanged free T levels over time. Among controls, no significant changes in basal androgens were observed over the time of the study. There were no significant differences in either the basal or ACTH-stimulated levels of DHEA, androstenedione, or F over the time of the study in either PCOS or control women. We conclude that the adrenocortical secretion of AAs or F in PCOS and control women remains stable over time, supporting the hypothesis that the adrenal response to ACTH may be an inherited trait. Alternatively, a decrease in DHEA-S levels over time was observed but only among PCOS patients whose initial levels of this metabolite were above the group median, suggesting that the activity of sulfotransferase in these patients may be up-regulated by factors other than those affecting adrenocortical biosynthesis and that such regulatory influences attenuate over time. PMID- 15531512 TI - Plasma ghrelin and resistin concentrations are suppressed in infants of insulin dependent diabetic mothers. AB - This study aimed to investigate 1) the effect of maternal diabetes mellitus on ghrelin, resistin, leptin, and insulin in term newborns; 2) the interrelationship of these metabolic hormones in the early postnatal period; and 3) the association of the hormones with anthropometric parameters at birth. A total of 120 term newborns were prospectively enrolled and categorized into three groups: 40 were infants of nondiabetic mothers (group N), 42 were infants born to mothers with gestational diabetes on low energy dietary treatment (group D), and 38 were infants born to mothers with preexisting or severe gestational diabetes who required exogenous insulin for stabilization of blood sugar during pregnancy (group I). Plasma ghrelin and resistin were significantly lower in group I than in either group N or group D infants (P < 0.048). Plasma ghrelin and subscapular skinfold thickness were significantly higher in female than in male infants [plasma ghrelin: median (interquartile range), 3.8 (3.0-4.8) vs. 3.0 (2.4-4.0) ng/ml in females and males, respectively; P = 0.003; subscapular skinfold thickness: 4.9 (4.2-5.6) vs. 4.6 (3.9-5.2) mm; P = 0.03]. In group N, plasma ghrelin was significantly, but negatively, associated with birth weight (r = 0.31; P = 0.05) and body length (r = -0.33; P = 0.04), whereas in group I, plasma ghrelin was negatively correlated with plasma resistin (r = -0.37; P = 0.02). Plasma ghrelin and resistin are suppressed in infants of insulin-dependent diabetic mothers, suggesting that the metabolic hormonal system is probably operational in fetal and early postnatal life. A low circulating ghrelin concentration may be advantageous to these infants, because a reduction in appetite may prevent excessive weight gain postnatally and counterbalances the in utero anabolic effect of hyperinsulinism in poorly controlled diabetic mothers. The suppressive effect of insulin on resistin may partially explain the excess accumulation of adipose tissue in infants of diabetic mothers by reducing the inhibitory effect of resistin on adipogenesis. Female infants have significantly higher plasma ghrelin levels than male infants, suggesting that sexual dimorphism exists in utero. This study has also shown an association between some of the metabolic hormones in specific groups of infants and thus suggests that these hormones could have interacted in utero to regulate growth and fat storage during this critical period. PMID- 15531513 TI - Late life metabolic syndrome, early growth, and common polymorphism in the growth hormone and placental lactogen gene cluster. AB - Low rates of fetal and infant growth are associated with the metabolic syndrome and cardiovascular disease in later life. We investigated common genetic variation in the GH-CSH gene cluster on chromosome 17q23 encoding GH, placental lactogens [chorionic somatomammotropins (CSH)], and placental GH variant in relation to fetal and infant growth and phenotypic features of the metabolic syndrome in subjects aged 59-72 yr from Hertfordshire, UK. Allele groups T, D1, and D2 of a locus herein designated CSH1.01 were examined in relation to GH-CSH single nucleotide polymorphisms and to specific phenotypes. Average birth weights were similar for all genotype groups. Men with T alleles were significantly lighter at 1 yr of age, shorter as adults, and had higher blood pressures, fasting insulin (T/T 66% higher than D2/D2) and triglyceride concentrations, and insulin and glucose concentrations during a glucose tolerance test. Birth weight and 1-yr weight associations with metabolic syndrome traits were independent of the CSH1.01 effects. Common diversity in GH-CSH correlates with low 1-yr weight and with features of the metabolic syndrome in later life. GH-CSH genotype adds substantially to, but does not account for, the associations between low body weight, at birth and in infancy, and the metabolic syndrome. PMID- 15531514 TI - Interleukin-6 regulates human adipose tissue lipid metabolism and leptin production in vitro. AB - Adipose tissue IL-6 expression is increased in obesity and is a strong predictor of abnormalities in adipocyte and systemic metabolism. We used adipose tissue organ culture to test the direct effects of IL-6 on leptin expression, lipolysis, and lipoprotein lipase activity. To assess possible interactions with the hormonal milieu, IL-6 effects were tested in the presence or absence of insulin and/or glucocorticoid [dexamethasone (dex)]. Because omental (Om) and abdominal sc depots differ in IL-6 expression, their responses to exogenous IL-6 were compared. Although IL-6 had no significant effects under basal conditions, culture with the combination of IL-6 and dex, compared with dex alone, for 2 d increased leptin in both depots [+95 +/- 30% (sc) and +67 +/- 19% (Om), P < 0.01]; IL-6 did not affect leptin production when added in the presence of insulin. Culture with IL-6 in the absence of hormones moderately increased lipolysis during culture in both sc and Om [+79 +/- 23% (sc) and +26 +/- 9% (Om), each P < 0.01]. IL-6 markedly reduced the high levels of lipoprotein lipase activity in tissue cultured with insulin plus dex. We conclude that high local concentrations of IL-6 can modulate leptin production and lipid metabolism in human adipose tissue. PMID- 15531515 TI - Genetic analyses of the HRPT2 gene in primary hyperparathyroidism: germline and somatic mutations in familial and sporadic parathyroid tumors. AB - We investigated the involvement of the HRPT2 gene by loss of heterozygosity analysis and direct sequencing in a kindred with hyperparathyroidism-jaw tumor syndrome (HPT-JT) and three kindreds with familial isolated primary hyperparathyroidism (FIHP). Seven patients with sporadic parathyroid cancers and 35 with parathyroid adenomas with no family history of primary hyperparathyroidism or HPT-JT were also studied. A germline heterozygous substitution G to A was found in the donor splice site of intron 1 in one of the three FIHP families. No mutations were identified in the HPT-JT kindred. A somatic HRPT2 mutation was found in four of seven patients with parathyroid cancers, two of which were unreported frameshift mutations (195insT and 195insA) in exon 2. Consistent with recent findings, two of seven patients with sporadic parathyroid cancer had germline mutations. Four adenomas showed loss of heterozygosity at HRPT2, whereas a somatic HRPT2 mutation was found in one. In conclusion, we provide additional evidence for a strong association between HRPT2 gene mutations and sporadic parathyroid cancer. The finding that two of the seven patients with sporadic parathyroid cancer carried an HRPT2 germline mutation suggests that they might have occult HPT-JT. Our results also confirm the need for testing HRPT2 gene in FIHP families. PMID- 15531516 TI - Endothelial dysfunction in young women with polycystic ovary syndrome: relationship with insulin resistance and low-grade chronic inflammation. AB - Women with polycystic ovary syndrome (PCOS) carry a number of cardiovascular risk factors. Cardiovascular morbidity is elevated even in young women with PCOS. Low grade chronic inflammation, reflected in elevated levels of serum C-reactive protein (CRP) and endothelial dysfunction have recently been linked to development of atherosclerosis. We compared high-sensitivity (hs)CRP concentrations and endothelium dysfunction in 37 women with PCOS and 25 control subjects matched as a group for age and body mass index (BMI). Arterial endothelium and smooth muscle function was measured by examining brachial artery responses to endothelium-dependent and endothelium-independent stimuli. Serum LH, testosterone, androstenedione, and fasting insulin levels were significantly higher in the PCOS group than the control group. The PCOS group was more insulin resistant than age- and BMI-matched control women. CRP concentrations were higher in PCOS women than the healthy control group (0.25 vs. 0.09 mg/dl). hsCRP concentrations were correlated with BMI, insulin sensitivity indices (homeostasis model assessment and quantitative insulin sensitivity check index), and endothelium-dependent vasodilation. The groups were well matched for baseline brachial artery diameter. There was a significant difference in endothelium dependent (flow- mediated dilation) and endothelium-independent (sublingual nitroglycerin) vascular responses between the women with PCOS and the normal healthy control group (P = 0.002 and P = 0.01, respectively). Endothelium dependent vasodilation was correlated with hsCRP concentrations and insulin resistance. In conclusion, this study is the first to demonstrate increased levels of hsCRP, endothelial dysfunction, and the relation with insulin resistance in young and normal-weight women with PCOS. Clinical strategies aimed at reducing insulin resistance may prevent early atherosclerosis in women with PCOS. PMID- 15531517 TI - Human thyroid phenol sulfotransferase enzymes 1A1 and 1A3: activities in normal and diseased thyroid glands, and inhibition by thyroid hormones and phytoestrogens. AB - Sulfation by sulfotransferase enzymes (SULTs) is an important pathway for the metabolism of thyroid hormones and phytoestrogens. Intrathyroidal SULTs may contribute to the processing of thyroid hormones for the reutilization of iodide. SULT1A1 and SULT1A3 activities were identified in normal and diseased human thyroid glands. Biochemical properties that included apparent K(m) values, thermal stabilities, and responses to inhibitors were characterized in a normal human thyroid high speed supernatant pool. Apparent K(m) values for SULT1A1 and SULT1A3 activities with the model substrates p-nitrophenol and dopamine were 0.58 +/- 0.04 and 11.3 +/- 1.3 microm, respectively. Activities of SULT1A1 and SULT1A3 determined in individual normal thyroid (n = 35), nodular goiter (n = 26), and autoimmune thyroid disease (n = 25) glands were 0.34 +/- 0.06, 0.52 +/- 0.09, and 0.82 +/- 0.19 U/mg protein for SULT1A1, respectively, and 0.22 +/- 0.04, 0.21 +/- 0.04, and 0.48 +/- 0.11 U/mg protein for SULT1A3, respectively. Both SULT activities in autoimmune thyroid disease glands were significantly higher than those in normal thyroids. Only 3,3'-diiodothyronine (3,3'-T(2)) and the phytoestrogen daidzein served as substrates for the normal thyroid SULT activities, yet each thyroid hormone and phytoestrogen tested were found to inhibit thyroid SULT1A1 and SULT1A3 activities. The preference of thyroid gland SULT activities for 3,3'-T(2) suggests that sulfation may enhance degradation of intrathyroidal 3,3'-T(2) for iodide reutilization. Inhibition of these SULT activities by the exogenous phytoestrogens daidzein and genistein, with a potential decrease in iodide reutilization, presents another mechanism through which these compounds may adversely affect human thyroid function. PMID- 15531518 TI - Prolonged mild hyperglycemia induces vagally mediated compensatory increase in C Peptide secretion in humans. AB - Experimentally induced prolonged hyperglycemia increases insulin release in humans, and in animals has been demonstrated to increase vagal efferent activity. The objective of the present experiment was to determine whether in humans, the compensatory increase in insulin release in response to short-term mild hyperglycemia is mediated by an induction of vagal efferent activity. Lean male subjects (n = 11; body mass index, 23.6 +/- 0.8 kg/m(2)) underwent a frequently sampled iv glucose tolerance test (FSIGT) to determine B cell function and insulin sensitivity. Subjects were then tested under four conditions over 4 months. Subjects were infused for 48 h with either glucose (15% dextrose at 200 mg/m(2).min) or saline (50 ml/h). Three hours after termination of the infusion, an FSIGT was administered in the presence of either saline or atropine (0.4 mg/m(2) bolus: 0.4 mg/m(2).h). Glucose (117 +/- 14 vs. 98 +/- 5 mg/dl) and insulin (49.5 +/- 10 vs. 23 +/- 5 muU/ml) levels were significantly elevated during the 48-h glucose infusion compared with those during saline treatment. Forty-eight-hour glucose infusions increased insulin and C-peptide levels during the FSIGT. When the FSIGT was conducted in the presence of atropine after glucose infusion, C-peptide levels were significantly attenuated during the period of endogenous insulin secretion (0-20 min; 31.8 +/- 13 vs. 39.2 +/- 11.9, atropine vs. no atropine) and exogenous insulin administration [20-40 min; 18.8 +/- 10.8 vs. 31.6 +/- 12.9., atropine vs. no atropine; F(3,9) = 4.99; P < 0.026]. A significant negative correlation was found between the repression of C-peptide by muscarinic blockade and the magnitude of the C-peptide response to the glucose infusion (r = 0.60; P < 0.045). Insulin AUC was not significantly altered by the presence of muscarinic blockade. In summary, we found that prolonged mild hyperglycemia results in a compensatory increase in C-peptide secretion, which is partially mediated by an induction in vagal efferent activity. PMID- 15531519 TI - Glucocorticoids stimulate the expression of 11beta-hydroxysteroid dehydrogenase type 2 in cultured human placental trophoblast cells. AB - Proper glucocorticoid exposure in utero is vital for normal fetal organ growth and maturation. The placental enzyme, 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), plays a pivotal role in controlling fetal exposure to high levels of maternal glucocorticoid by converting cortisol into its inactive metabolite, cortisone. The present study was designed to determine whether glucocorticoids auto-regulate 11beta-HSD2 in the human placenta using cultured trophoblast cells as a model system. Trophoblasts were isolated from uncomplicated term placentas and treated with glucocorticoids. The synthetic glucocorticoid dexamethasone increased 11beta-HSD2 activity in a time- and concentration-dependent manner; this effect was accompanied by a corresponding increase in 11beta-HSD2 mRNA. Furthermore, the glucocorticoid receptor antagonist, RU-486, abrogated the dexamethasone-induced increase in 11beta-HSD2 activity, suggesting that the effect of dexamethasone is mediated through the glucocorticoid receptor. Results from transient transfection and mRNA decay experiments indicate that the glucocorticoid-induced increase in 11beta-HSD2 expression is mediated at both the transcriptional and posttranscriptional levels. In conclusion, the present study demonstrates that in cultured human trophoblasts, 11beta-HSD2 is subject to auto regulation by glucocorticoids. If this occurs in the human placenta in vivo, the glucocorticoid-induced up-regulation of placental 11beta-HSD2 would represent an important safeguard mechanism by which the fetus may be protected from detrimental exposure to elevated levels of maternal glucocorticoids. PMID- 15531520 TI - Increased release of cervical nitric oxide in spontaneous abortion before clinical symptoms: a possible mechanism for preabortal cervical ripening. AB - Nitric oxide (NO) affects cervical ripening. We studied cervical NO release in women with nonviable pregnancy before signs of abortion. Women with missed abortion (n = 56), blighted ovum (n = 36), or tubal pregnancy (n = 7) were selected by means of vaginal ultrasonographic examination from a population seeking early pregnancy termination; 140 women with amenorrhea-matched normal gestation were studied as controls. Cervical fluid samples were assessed for NO metabolites (Nox) by means of Griess reaction. Cervical fluid Nox was more often detectable in women with missed abortion (90%) and blighted ovum (87%) than in the control women (55%; P = 0.01), and Nox levels in women with missed abortion [median, 59.4 mumol/liter; 95% confidence interval (CI), 30.3-81.8] and blighted ovum (25.6 mumol/liter; 95% CI, 14.1-53.0) were 14 and 6 times higher (P < 0.001 and P = 0.002, respectively) than in the control group (4.3 mumol/liter; 95% CI, <3.8 to 6.4). Nox levels in women with tubal pregnancy were normal. In women with nonviable pregnancy, the lower the level of progesterone, expressed as a percentage of that in the control women, the higher (r = -0.69; P < 0.001) the level of cervical fluid Nox, and those with low pretreatment Nox levels failed to abort completely after mifepristone-misoprostol or expectant management more often (P = 0.04) than women with high Nox levels (28% vs. 4%); no such relationship was seen in the control group. Increased preabortal cervical NO release may contribute to cervical ripening and the onset of clinical abortion. PMID- 15531521 TI - Release of proinflammatory cytokines and 8-isoprostane from placenta, adipose tissue, and skeletal muscle from normal pregnant women and women with gestational diabetes mellitus. AB - The aim of this study was to 1) profile the basal release of TNF-alpha, IL-6, IL 8, and 8-isoprostane (a marker of oxidative stress); and 2) investigate the effect of stimulation on the release of cytokines from sc adipose tissue and skeletal muscle from normal pregnant women and women with gestational diabetes mellitus (GDM). Placenta, sc adipose tissue, and skeletal muscle were incubated in the absence (control) or presence of lipopolysaccharide (LPS; 10 microg/ml), TNF-alpha (10 ng/ml), IL-6 (10 ng/ml), or IL-8 (10 ng/ml). After an 18-h incubation, the medium was collected, and the release of TNF-alpha, IL-6, IL-8, and 8-isoprostane was quantified by ELISA. In all three tissues, 8-isoprostane release was greater in women with GDM, and stimulation with LPS increased 8 isoprostane release from adipose and skeletal muscle, but not placenta, obtained from women with GDM. However, in tissues obtained from normal pregnant women, LPS stimulation increased 8-isoprostane release in placenta and had no effect in adipose tissue and skeletal muscle. Their was no difference in the release of TNF alpha, IL-6, and IL-8 from placenta, adipose tissue, and skeletal muscle obtained from normal pregnant women and women with GDM. Stimulation of placenta, adipose tissue, and skeletal muscle with LPS and TNF-alpha resulted in greater release of IL-6 and IL-8, whereas only LPS increased TNF-alpha release from all three tissues. The data presented in this study demonstrate that there is a differential release of 8-isoprostane from fetal (placenta) and maternal (adipose tissue and skeletal muscle) tissues obtained from normal pregnant women and women with GDM. These data are consistent with the hypothesis that oxidative stress may be involved in the progression and/or pathogenesis of GDM. PMID- 15531522 TI - Blood ionized calcium is associated with clustered polymorphisms in the carboxyl terminal tail of the calcium-sensing receptor. AB - Blood ionized calcium (iCa) is a quantitative trait subject to genetic influence. iCa is maintained in a narrow range through the action of the calcium-sensing receptor (CASR) controlling PTH secretion and calcium excretion. A CASR single nucleotide polymorphism (SNP) prevalent in Caucasian populations (A986S) has shown significant association with iCa in a cohort of young women, but association with the neighboring SNPs, R990G and Q1011E, has not been examined. We studied 377 unrelated adults (184 men and 193 women) recruited as healthy adults from a blood donor clinic. The subjects were not taking any medications, nor did they have disorders of calcium metabolism. Relative frequencies for the CASR 986S, 990G, and 1011E minor alleles were 24%, 4%, and 3% respectively. At the A986S locus, subjects with the AA genotype had significantly lower iCa (P = 0.0001) than subjects with one or two S alleles (mean +/- se, 1.221 +/- 0.003 vs. 1.239 +/- 0.003 mmol/liter). For the R990G site, subjects with the RR genotype had higher iCa than those with one copy of the 990G allele (1.230 +/- 0.002 vs. 1.213 +/- 0.007 mmol/liter; P = 0.032). With respect to the 1011 locus, iCa was lower in QQ genotype subjects than in the QE group (1.227 +/- 0.002 vs. 1.255 +/- 0.008 mmol/liter; P = 0.002). After resolution of phase for the doubly heterozygous subjects, analysis was conducted on haplotypes across all three loci. As expected, subjects with SRQ and ARE haplotypes are relatively hypercalcemic, and those with AGQ are hypocalcemic, relative to subjects with the common ARQ haplotype. Multiple regression analysis with clinical covariates (age, sex and menopausal status, creatinine, and PTH) showed that 16.5% of the total variance in iCa may be explained, and the seven CASR haplotypes contribute significantly (P < 0.0001) and substantially (49.1% of the explained variance) to the model, with the following corrected iCa means: ARQ/AGQ, 1.21 +/- 0.01; ARQ/ARQ, 1.22 +/- 0.01; ARQ/SRQ, 1.24 +/- 0.01; SRQ/AGQ, 1.24 +/- 0.03; SRQ/SRQ, 1.25 +/- 0.01; ARQ/ARE, 1.25 +/- 0.01; and SRQ/ARE, 1.27 +/- 0.01. Our data confirm the association between iCa and the A986S locus and suggest that R990G and Q1011E are also predictive. Given the significant between-population variations in frequency of variant alleles in this CASR SNP cluster, tri-locus haplotyping may prove to be more informative in studies of association between variation in CASR and disease. PMID- 15531523 TI - Regulation of 15-hydroxyprostaglandin dehydrogenase (PGDH) gene activity, messenger ribonucleic acid processing, and protein abundance in the human chorion in late gestation and labor. AB - The prostaglandin (PG)-inactivating enzyme 15-hydroxyprostaglandin dehydrogenase (PGDH) is highly expressed in the chorion leave. To assess the involvement of PGDH in the regulation of intrauterine PG levels, we have determined the mechanisms that control chorionic PGDH expression in women at term and preterm labor. PGDH gene activity decreased at term and during normal labor. PGDH mRNA abundance also decreased at term due to changing splice variant distribution. Gene activity predicted PGDH mRNA abundance preterm and after normal labor, but not at term before labor. PGDH mRNA decayed rapidly in cultured tissues and was stabilized by transcriptional arrest. PGDH protein levels varied without being significantly different between the patient groups. PGDH mRNA levels predicted PGDH protein levels at term, but not preterm and after labor. PGDH gene activity, mRNA variant, and immunoreactive protein levels were not different between the preterm labor and preterm not in labor groups. Thus, PGDH mRNA is transiently down-regulated before term labor by a posttranscriptional mechanism(s). Protein turnover controls PGDH protein abundance at preterm and after normal labor. At term, PGDH protein levels become dependent on the rapidly turning over PGDH mRNA. This may allow rapid changes in PGDH protein abundance and uterotonic PG concentrations promoting labor. PMID- 15531524 TI - Extreme elevation of intrasellar pressure in patients with pituitary tumor apoplexy: relation to pituitary function. AB - The dominant mechanism for hypopituitarism and hyperprolactinemia commonly observed in patients with pituitary macroadenomas was postulated to be increased intrasellar pressure (ISP) caused by the slow and gradual expansion of adenomas within the sella turcica. Hemorrhagic infarction of adenomas (pituitary tumor apoplexy) is associated with a rapid, rather than gradual, increase in intrasellar contents. The impacts of the sudden increase in intrasellar contents on ISP and pituitary function are unknown. ISP and pituitary function were determined in 13 patients with pituitary tumor apoplexy who had surgical decompression within 1 wk of symptoms' onset. ISP measurements were remarkably high (median, 47 mm Hg), whereas serum prolactin (PRL) concentrations were generally low (median, 3.5 microg/liter). There was an inverse correlation (r = 0.76; P < 0.01) between ISP measurements and serum PRL concentrations. Postoperatively, partial recovery or maintenance of pituitary function was noted in seven of 13 patients. These seven patients had higher (P = 0.013) serum PRL levels (9.3 +/- 7.4 microg/liter) and lower (P < 0.001) ISP measurements (35.9 +/ 7.3 mm Hg) than the respective values in the remaining six with persistent postoperative hypopituitarism (1.6 +/- 0.6 microg/liter and 55.9 +/- 2.4 mm Hg, respectively). The low serum PRL levels in patients with tumor apoplexy suggested that ischemic necrosis of the anterior pituitary resulting from sudden and extreme elevation of ISP was commonly observed in this setting. A normal or elevated serum PRL level in patients with non-PRL-secreting macroadenomas indicates the presence of viable pituitary cells and the high likelihood of postoperative recovery of pituitary function. PMID- 15531525 TI - A single-base mutation in the peroxisome proliferator-activated receptor gamma4 promoter associated with altered in vitro expression and partial lipodystrophy. AB - Familial partial lipodystrophy (FPLD) results from coding sequence mutations either in LMNA, encoding nuclear lamin A/C, or in PPARG, encoding peroxisome proliferator-activated receptor gamma (PPARgamma). The LMNA form is called FPLD2 (MIM 151660), and the PPARG form is called FPLD3 (MIM 604367). We now report a 21 yr-old female with FPLD and no coding sequence mutations in either LMNA or PPARG. She was heterozygous for a novel A>G mutation at position -14 of intron B upstream of PPARG exon 1 within the promoter of the PPARgamma4 isoform. Her less severely affected father, who had features of the metabolic syndrome and a paucity of limb and gluteal fat, was also heterozygous for -14A>G. This mutation was absent among 600 alleles from normal Caucasians. A minimal promoter sequence bearing the mutation had significantly reduced promoter activity when used to drive reporter expression in in vitro expression in two cell lines, compared with the wild-type sequence. This is the first report of a human mutation in the promoter of a PPARgamma isoform. Because the mutation affects PPARgamma4 expression and is associated with FPLD, this implies that PPARgamma4 might be important for fat depot distribution and metabolism in vivo. PMID- 15531526 TI - Increased expression of type I 17beta-hydroxysteroid dehydrogenase enhances in situ production of estradiol in uterine leiomyoma. AB - Expression of 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) was compared between leiomyoma and myometrium. Cytosolic fractions from leiomyoma homogenate displayed 5-fold higher activity (estrone to estradiol), compared with surrounding myometrium (n = 6, P < 0.05), whereas microsomal fractions showed no difference. Oxidative activity (estradiol to estrone) did not differ between leiomyoma and myometrium. Levels of mRNA for 17beta-HSDs were then measured using real-time PCR techniques. Among the eight different types of 17beta-HSDs (types 1 5, 7, 8, and 10), type 1 was the only enzyme displaying differential expression between leiomyoma and myometrium. Mean concentration of type 1 17beta-HSD mRNA was 4-fold higher in leiomyoma than in surrounding myometrium (n = 20, P < 0.05). Type 1 transcript levels correlated significantly with reductive activity in individual samples (n = 6, P < 0.05). Northern blot analysis of leiomyoma and myometrium tissues detected 2.3- and 1.0-kb transcripts of type 1 enzyme, whereas the major 1.3-kb transcript for 17beta-HSD in placenta-derived JEG-3 cells was not detected. None of the factors increasing mRNA levels for type 1 enzyme in placenta increased mRNA levels in leiomyoma. These results indicate that leiomyoma tissues overexpress type 1 17beta-HSD, resulting in high conversion of estrone to estradiol. In situ expression of type 1 17beta-HSD may play a role in self-supported growth of leiomyoma cells. PMID- 15531527 TI - Familial PAX8 small deletion (c.989_992delACCC) associated with extreme phenotype variability. AB - The PAX8 gene, mapped on 2q12-q14, encodes for a transcription factor involved in thyroid cell proliferation and differentiation. Five mutations in PAX8 have been so far described in both sporadic and rare familial forms of thyroid dysgenesis with proposed autosomal dominant inheritance, all associated with thyroid hypoplasia and/or dysfunction. Fifty-four subjects with congenital hypothyroidism detected during neonatal screening and associated with an ultrasound or scintiscan picture of thyroid dysgenesis were investigated for PAX8 mutations. The entire PAX8 coding region with exon-intron boundaries was amplified from genomic DNA, and a mutational screening was performed by denaturing HPLC followed by direct sequencing when denaturing HPLC elution abnormalities appeared. A new heterozygous deletion (c.989_992delACCC) in exon 7 causing a frameshift with premature stop codon after codon 277 was identified in a subject with thyroid hypoplasia. This mutation is the only one so far identified that lies outside the paired domain. The predicted mutant protein completely lacks the C-terminal region but contains the paired box, octapeptide, and homeodomain. It retains the ability to bind a paired-domain sequence in vitro but is transcriptionally inactive. These results provide evidence that the C-terminal region is essential for transcriptional activity. The new mutation has been inherited from the completely euthyroid mother. It was also present in a brother with slightly elevated TSH only. Thus, it is associated with thyroid dysgenesis in the proband and both euthyroidism and compensated hypothyroidism in her family. This suggests that other factors/genes may modulate phenotypic expression. PMID- 15531528 TI - Epitope-restricted 65-kilodalton glutamic acid decarboxylase autoantibodies among new-onset Sardinian type 2 diabetes patients define phenotypes of autoimmune diabetes. AB - The 65-kDa glutamic acid decarboxylase (GAD65) autoantibodies (GAD65Abs), commonly found in type 1 diabetes mellitus (T1DM) patients, are also found at lower frequencies in type 2 diabetes mellitus (T2DM) patients. GAD65Abs in T1DM patients are epitope specific, in contrast to those found in other GAD65Ab positive individuals, including T2DM patients. Our aim was to assess whether epitope-specific GAD65Abs, or the additional presence of islet antigen 2 (IA-2) autoantibodies, better define T1DM phenotypes among T2DM patients. GAD65 and IA-2 autoantibodies were analyzed in 1436 Sardinian subjects classified with T2DM and in 384 nondiabetic patient controls. Autoantibody binding specificity to the N terminal, middle (M), and C-terminal (C) portions of the GAD65 molecule was evaluated. Among the T2DM patients, 5.1% had GAD65 (P < 0.001) and 2.4% had IA-2 autoantibodies, compared with 1.3 and 1.6%, respectively, among the controls. GAD65Ab-positive T2DM patients with M+C (epitope-specific) reactivity were found to have the lowest body mass index (P < 0.001), followed by GAD65Ab/IA-2Ab positive patients (P < 0.01), and non-M+C-reactive (non-epitope-specific) patients (P < 0.02). In GAD65Ab-positive T2DM patients, c-peptide levels were lower in M+C-reactive compared with non-M+C-reactive patients. Sardinian T2DM patients with M+C-predominant GAD65Ab reactivity have clinical features more similar to those of T1DM patients. Thus, GAD65Ab epitope analysis may help to define T1DM phenotypes among newly diagnosed GAD65Ab-positive patients classified with T2DM. PMID- 15531529 TI - Expression and deoxyribonucleic acid-binding activity of the nuclear factor kappaB family in the human myometrium during pregnancy and labor. AB - In humans, the factors that govern the switch from myometrial quiescence to coordinated contractions at the initiation of labor are not well defined. The onset of parturition is itself associated with increases in a number of proinflammatory factors, many of which are regulated by the nuclear factor kappaB (NF-kappaB) family of transcription factors. The expression and DNA-binding activity of NF-kappaB in the myometrium during gestation and parturition were examined. Levels of c-Rel, p50, and p105 NF-kappaB species were dramatically reduced in pregnant myometrium compared with nonpregnant (NP) controls, whereas expression of the RelA subunit remained uniform. Importantly, during labor, expression of all subunits was observed to be significantly reduced in all myometrial samples studied relative to NP levels. Moreover, for RelA, c-Rel, and p50 subunits, there was a gradient of expression between laboring upper (corpus) and lower uterine segment myometrium. No RelB or p52 subunits could be detected. EMSAs identified changes in NF-kappaB subunit composition in the myometrium during pregnancy and labor, with p50 homodimers predominant in NP tissues being replaced with RelA:p50 heterodimers in pregnant and laboring samples. Significantly, RelA was observed to be phosphorylated at serine-536, implicating the involvement of the phosphatidylinositol-3-kinase/AKT pathway in NF-kappaB function in the myometrium. PMID- 15531530 TI - Large germline deletions of mitochondrial complex II subunits SDHB and SDHD in hereditary paraganglioma. AB - More than 30% of adrenal pheochromocytomas are hereditary. These neuroendocrine tumors are major components of three inherited cancer syndromes: multiple endocrine neoplasia type 2, von Hippel-Lindau disease (VHL), and pheochromocytoma/paraganglioma syndrome (PC/PGL). Germline mutations in RET; VHL; and SDHB, SDHC, and SDHD are associated with multiple endocrine neoplasia type 2, VHL, and PC/PGL, respectively. The majority (>70%) of hereditary extraadrenal PCs [catecholamine-secreting paragangliomas (PGL)] are accounted for by germline intragenic mutations in SDHB, SDHC, or SDHD. Therefore, a subset of hereditary PGL is not accounted for. Here we report two unrelated hereditary PGL families, one with a germline whole-gene deletion of SDHD (family 4194), the other a partial deletion of SDHB (family BRZ01). Although they were initially designated mutation negative for all of the PC-associated genes after PCR-based analysis, we suspected that a large deletion or rearrangement might be present. Genotyping around the PC-associated genes demonstrated that both families were consistent with linkage with one of these genes. Using fine structure genotyping and semiquantitative duplex PCR analysis, we identified an approximately 96-kb deletion spanning SDHD in family 4194 and an approximately 1-kb deletion involving the 5' end of SDHB in family BRZ01. Thus, including SDHB and SDHD deletion analysis could increase gene-testing sensitivity for PGL patients, which would aid in genetic counseling and management of patients and families. PMID- 15531531 TI - The insulin-like growth factor-II receptor gene is associated with type 1 diabetes: evidence of a maternal effect. AB - Susceptibility to type 1 diabetes (T1D) is a complex trait, involving several loci. One of these putative loci, insulin-dependent diabetes mellitus-8 (IDDM8) at 6q, has been found to be subject to parental effects, suggesting the involvement of an imprinted gene. IGF-II receptor (IGF2R), the best-studied imprinted gene in the IDDM8 region, encodes the IGF-2 receptor, a protein involved in many biological processes, including immune function and beta-cell regeneration. Mice express only the maternal allele. In humans, the molecular IGF2R imprint (maternal-specific methylation) is present, but it affects expression in only a small subset of individuals. To examine whether IGF2R might contribute to the IDDM8 effect, we examined transmission distortion at several single nucleotide polymorphisms (SNPs) in 404 parent-offspring trios. After correcting for multiple testing, significant distortion was found at only one silent SNP on exon 16 (P = 0.002). SNPs upstream and downstream showed weak linkage disequilibrium and no transmission distortion, localizing the association to a 53-kb block within IGF2R. Interestingly, the exon 16 SNP association was limited to maternally inherited alleles. SLC22A2 and SLC22A3, two genes downstream of IGF2R that are imprinted in the mouse, showed no T1D association. Thus, we present evidence that maternal alleles at an IGF2R polymorphism are associated with T1D. It is thus possible that at some tissue or developmental stage not yet examined, IGF2R is universally imprinted. PMID- 15531532 TI - Plasma levels of intact and degraded ghrelin and their responses to glucose infusion in anorexia nervosa. AB - Octanoylated ghrelin (1-28) (intact ghrelin) is rapidly and easily degraded to desoctanoyl forms or smaller fragments (degraded ghrelin). Plasma levels of intact and degraded ghrelin were examined in 30 patients with anorexia nervosa (AN) (body mass index, 8.81-22.4 kg/m(2)) and 16 age-matched healthy women using several assay methods. Plasma levels of ghrelin measured using immunocomplex transfer-enzyme immunoassay, which specifically detects intact ghrelin, were lower in AN than controls. Plasma ghrelin levels in AN measured using the active ghrelin ELISA kit, which is advertised as specifically detecting intact ghrelin, did not differ significantly from controls. Plasma levels of desoctanoyl ghrelin using the desacyl-ghrelin ELISA kit, N-terminus ghrelin using the ghrelin active RIA kit, and C-terminus ghrelin using the ghrelin total RIA kit were significantly higher in AN than controls, and displayed significant negative correlations with body mass index. Plasma levels of ghrelin determined using immunocomplex transfer-enzyme immunoassay or active ghrelin ELISA during iv glucose infusion were suppressed in both AN and controls, whereas plasma levels of degraded ghrelin levels were not significantly decreased in AN. Plasma levels of intact ghrelin are therefore not higher in AN than controls, whereas degraded forms of ghrelin are elevated in AN. Rapid suppression of plasma intact ghrelin, but not degraded ghrelin, occurs in AN in response to glucose infusion. The profiles of intact and degraded forms of ghrelin in plasma of AN patients differ from those of healthy women. PMID- 15531533 TI - Elevated pregnancy-associated plasma protein-a in sera from type 2 diabetic patients with hypercholesterolemia: associations with carotid atherosclerosis and toe-brachial index. AB - Pregnancy-associated plasma protein (PAPP)-A, a superfamily of metalloproteinase, has been implicated in acute coronary syndrome. We compared PAPP-A concentrations in sera from patients with type 2 diabetes with those in sera from age-matched control subjects and also investigated whether serum PAPP-A was associated with carotid intima-media wall thickness (IMT), an early marker of atherosclerosis, and indices of peripheral vascular disease in the diabetic patients. Serum PAPP-A was measured by an ELISA in 103 type 2 diabetic patients and 32 age-matched control subjects. All subjects were not pregnant. IMT was evaluated ultrasonographically in both common carotid arteries. As measures of peripheral vascular disease, we also determined the ankle-brachial index and toe-brachial index (TBI) for systolic blood pressure. Hypercholesterolemia was defined as a serum low-density lipoprotein-cholesterol concentration exceeding 3.6 mmol/liter or alternatively as a treatment with hydroxymethylglutaryl coenzyme A reductase inhibitor. Serum PAPP-A was significantly higher in diabetic patients than control subjects (P < 0.0001). In diabetic patients, serum PAPP-A correlated positively with serum total cholesterol (r = 0.289, P = 0.0041) and IMT (r = 0.315, P = 0.0017) and negatively with TBI (r = -0.294, P = 0.0039) but not ankle brachial index. Diabetic patients with hypercholesterolemia had higher PAPP-A concentrations than those without hypercholesterolemia [median (interquartile ranges): 8.37 (6.93, 11.6) vs. 7.29 (5.65, 9.21) mIU/liter; P = 0.0209]. Multivariate analysis identified only serum total cholesterol as an independent determinant of serum PAPP-A in patients with type 2 diabetes (partial coefficient 0.454, P = 0.020). In conclusion, serum PAPP-A concentrations were significantly elevated in diabetic patients with hypercholesterolemia and were associated positively with carotid atherosclerosis and negatively with TBI in type 2 diabetes. PMID- 15531534 TI - Decrease in luteinizing hormone pulse frequency during a five-hour peripheral ghrelin infusion in the ovariectomized rhesus monkey. AB - Ghrelin, a nutrition-related peptide secreted by the stomach, is elevated during prolonged food deprivation. Because undernutrition is often associated with a suppressed reproductive axis, we have postulated that increasing peripheral ghrelin levels will decrease the activity of the GnRH pulse generator. Adult ovariectomized rhesus monkeys (n = 6) were subjected to a 5-h iv human ghrelin (100- to 150-microg bolus followed by 100-150 microg/h) or saline infusion, preceded by a 3-h saline infusion to establish baseline pulsatile LH release. Blood samples were collected at 15-min intervals throughout the experiment. Ghrelin infusion increased plasma ghrelin levels 2.9-fold of baseline. Ghrelin significantly decreased LH pulse frequency (from 0.89 +/- 0.07/h in baseline to 0.57 +/- 0.10/h during ghrelin infusion; P < 0.05, mean +/- sem), whereas LH pulse frequency remained unchanged during saline treatment. LH pulse amplitude was not affected. Ghrelin also significantly stimulated both cortisol and GH release, but had no effect on leptin. We conclude that ghrelin can inhibit GnRH pulse activity and may thereby mediate the suppression of the reproductive system observed in conditions of undernutrition, such as in anorexia nervosa. Ghrelin also activates the adrenal axis, but the relevance of this to the inhibition of GnRH pulse frequency remains to be established. PMID- 15531535 TI - Tissue factor and CCL2/monocyte chemoattractant protein-1 released by human islets affect islet engraftment in type 1 diabetic recipients. AB - Islet survival in the early posttransplantation period is likely to be influenced by inflammatory events in and around islets. Twenty-seven human islet preparations were transplanted by 24 infusions into 14 patients with brittle type 1 diabetes under the Edmonton protocol. Patients were monitored for their coagulation [cross-linked fibrin degradation products (XDPs)] and liver function test [aspartate and alanine aminotransferase (AST and ALT)] as markers of early posttransplant complications, and these were correlated with in vitro islet number, purification, volume, monocyte-chemoattractant protein-1 (CCL2/MCP-1) and tissue factor (TF) islet release. Consistent with activation of coagulation pathways and hepatic damage, serum XDP values increased early after 11 infusions and transaminase after 13 of 24 infusions. TF and CCL2/MCP-1 were detected in supernatants of 21 and 22 islet preparations, respectively. Serum XDP peak values were correlated with TF/equivalent islets (EI) (r(2)=0.26, P = 0.001) and CCL2/MCP-1/EI (r(2) = 0.42; P < 0.001); serum transaminase areas under the curve in the first week posttransplantation were correlated with CCL2/MCP-1/EI (r(2) = 0.55; P < 0.001 for ALT and r(2) = 0.51; P = 0.001 for AST) and TF/EI (r(2) = 0.31; P = 0.002 for ALT, and r(2) = 0.36; P = 0.002 for AST). These data suggest that reducing the islet proinflammatory state may be a means to reduce the early posttransplant complications and perhaps improve islet engraftment. PMID- 15531536 TI - Behavioral, adrenal, and sympathetic responses to long-term administration of an oral corticotropin-releasing hormone receptor antagonist in a primate stress paradigm. AB - CRH is a main regulator of the stress response. This neuropeptide and its specific receptors, CRHR-1 and CRHR-2, are disseminated throughout the central nervous system. There is a significant interspecies difference in the distribution of CRHR within the central nervous system. CRH-R1 antagonists may attenuate stress-related behavior in rats without compromising adrenal function, but few studies have addressed the same question in higher mammals. Antalarmin (AA) is a specific CRHR-1 antagonist suitable for oral administration. Social separation is a potent stressor for rhesus monkeys. Therefore, we sought to investigate the hormonal responses to chronic administration of AA using a primate stress model. Eight preadolescent (4-6 kg) male rhesus monkeys received AA (20 mg/kg.d) or placebo (PBO) orally. All animals were on a regular day/light cycle and were fed with standard monkey chow daily. The study (114 d) was comprised of the following consecutive phases: adaptation, baseline, separation (stress), recovery, and cross-over. During social separation, solid panels separated the individuals. Cerebrospinal fluid (CSF) and femoral venous blood samples were obtained once a week on the fourth day of separation under ketamine anesthesia. Serum samples were also obtained 1 and 2 h after separation. CSF samples were assayed for CRH, AA, norepinephrine (NE) and epinephrine (EPI). Plasma was assayed for ACTH, cortisol, NE, and EPI. AA was detected in the plasma of each monkey while they were taking the active drug and in none of the animals on PBO. Among the behaviors assessed, environmental exploration, a behavior inhibited by stress, was increased during AA administration. However, AA at this dose did not affect other anxiety-related behavioral end points, including self directed behavior, vocalization, or locomotion. We also observed that: 1) ACTH decreased between adaptation and baseline, indicating that the animals had adjusted to the novel environment; 2) ACTH and cortisol increased significantly after social separation, indicating that social separation was an adequate model for acute stress; 3) NE and EPI increased significantly during acute stress in the AA and PBO groups (P < 0.005, NE; P < 0.001, EPI); 4) after chronic stress, by d 4 of separation, ACTH levels were no longer significantly different from baseline, and NE and EPI remained slightly elevated when compared with baseline (P < 0.05, NE; P < 0.01, EPI); and 5) all the animals remained healthy and gained the expected weight during the study. In summary, oral chronic administration of a specific CRH-R1 antagonist to rhesus monkeys does not blunt the sympathoadrenal response to stress while increasing environmental exploration, a behavior that is normally suppressed during stressful events. Taken together, these findings suggest that CRHR-1 antagonists may be a valid treatment for stress-related disorders. PMID- 15531537 TI - The effects of the ACE gene insertion/deletion polymorphism on glucose tolerance and insulin secretion in elderly people are modified by birth weight. AB - The I allele of an insertion/deletion (I/D) polymorphism in the angiotensin converting enzyme gene (ACE) appears to be protective against the complications of type 2 diabetes. Low birth weight, a marker of an adverse intrauterine environment, is associated with higher rates of type 2 diabetes. We examined whether the ACE I/D polymorphism could explain or modify the association between low birth weight and adulthood glucose tolerance. We measured plasma glucose and insulin concentrations after an oral glucose challenge in a group of 423 men and women, ages 65-75 yr, with measurements at birth recorded. The presence of the I allele was associated with shorter duration of gestation (P = 0.006) and, relative to gestational age, higher birth weight (P = 0.008) and length (P = 0.02). The I allele was associated with lower glucose at 120 min (P = 0.04) and a greater insulin response (P = 0.03 for insulin at 30 min and P = 0.06 for insulin area under the curve) to a standard oral glucose tolerance test. However, the associations between the ACE genotype and adulthood insulin secretion were only present in people with low birth weight (P for interaction birth weight * ACE genotype on insulin at 30 min = 0.003 and on insulin area under the curve = 0.05). The ACE I allele is associated with shorter duration of gestation and higher birth weight. The association between the presence of the ACE I allele and increased indices of adult insulin secretion is confined to subjects with low birth weight. We suggest that these findings reflect interactions between genotype and intrauterine environment with resulting changes in gene expression. PMID- 15531538 TI - Gene expression profiles and structural/functional features of the peri implantation endometrium in natural and gonadotropin-stimulated cycles. AB - It has been speculated that controlled ovarian hyperstimulation (COH), as performed during in vitro fertilization therapy, may negatively affect embryo implantation. The objective of this prospective and randomized study was to investigate gene expression profiles of the human endometrium during the window of implantation of gonadotropin-stimulated COH cycles compared with temporally matched natural cycles (d 21). Analysis was performed with high-density oligonucleotide microarrays. In addition, other structural and functional features of the endometrium were investigated. Results corroborated that COH cycles depicted advancement of pinopodes appearance, histological features, and steroid receptor down-regulation when compared with natural cycles. These changes were associated with significant, albeit small, variations in gene expression (18 genes/expressed sequence tags and -1.55- to +3.40-fold changes). Second, there were significant changes in gene expression when comparing cycles using a GnRH agonist vs. a GnRH antagonist (13 genes/expressed sequence tags and +1.42- to +2.10-fold changes). This is the first attempt to elucidate gene expression profiles of the endometrium during COH cycles. The observed differences in gene expression in COH cycles using state-of-the-art protocols may not have a major functional impact on embryo implantation. PMID- 15531539 TI - Erosion of endogenous testosterone-driven negative feedback on pulsatile luteinizing hormone secretion in healthy aging men. AB - The present study tests the intuition that successful aging in men is marked by: 1) impaired feedforward by endogenous LH concentrations (con) of testosterone (Te) secretion (sec); and/or 2) attenuated feedback by unmanipulated Te con of LH sec. The goal was to assess both implicit linkages analytically without disrupting normal pathway coupling. This strategy required: 1) assay of paired LH and Te con sampled every 10 min for 24 h in 13 older (O) (ages 60-78 yr) and 13 young (Y) (ages 18-30 yr) men; 2) deconvolution-based estimation of LH and Te sec rates; 3) lag-specific cross-correlation analyses of the relationships between LH and Te con and sec; and 4) statistical contrasts by age stratum. Salient outcomes were: 1) O and Y men maintain comparable LH con drive of Te sec, viz maximal r = +0.51 and r = +0.52, respectively, at an optimal time lag of 50 min (both P < 0.001 against random LH and Te associations); 2) elderly subjects exhibit reduced Te con inhibition of LH sec [minimal r = -0.008 (O) vs. r = -0.10 (Y), P < 0.01 at a time lag of 40 min]; 3) mean (24-h) LH con do not differ by age; and 4) molar Te/sex hormone-binding globulin con are lower in the elderly than in Y individuals (P < 0.01).In conclusion, noninvasive analyses predict that attenuation of endogenous Te feedback restraint on the hypothalamo-pituitary unit may be an early biological marker of adaptive changes in the GnRH-LH-Te ensemble axis in the healthy O male. PMID- 15531540 TI - Leptin levels are dependent on sleep duration: relationships with sympathovagal balance, carbohydrate regulation, cortisol, and thyrotropin. AB - Sleep plays an important role in energy homeostasis. The present study tests the hypothesis that circulating levels of leptin, a hormone that signals energy balance to the brain, are influenced by sleep duration. We also analyzed associations between leptin and sympathovagal balance, cortisol, TSH, glucose, and insulin under different bedtime conditions. Twenty-four-hour hormonal and glucose profiles were sampled at frequent intervals, and sympathovagal balance was estimated from heart rate variability in 11 subjects studied after 6 d of 4-h bedtimes (mean +/- sem of sleep duration during last 2 d: 3 h and 49 +/- 2 min) and after 6 d of 12-h bedtimes (sleep: 9 h and 03 +/- 15 min). A study with 8-h bedtimes was performed 1 yr later (sleep: 6 h and 52 +/- 10 min). Caloric intake and activity levels were carefully controlled in all studies. Mean levels, maximal levels, and rhythm amplitude of leptin were decreased (-19%, -26%, and 20%, respectively) during sleep restriction compared with sleep extension. The decrease in leptin levels was concomitant with an elevation of sympathovagal balance. The effects of sleep duration on leptin were quantitatively associated with alterations of the cortisol and TSH profiles and were accompanied by an elevation of postbreakfast homeostasis model assessment values. Measures of perceived stress were not increased during sleep restriction. During the study with 8-h bedtimes, hormonal and metabolic parameters were intermediate between those observed with 4-h and 12-h bedtimes. In conclusion, sleep modulates a major component of the neuroendocrine control of appetite. PMID- 15531541 TI - Biethnic comparisons of autosomal genomic scan for loci linked to plasma adiponectin in populations of Chinese and Japanese origin. AB - Adiponectin is secreted by adipocytes and is thought to have insulin-sensitizing and antiatherogenic effects. Two previous genome scans for plasma adiponectin have identified different regions for European and Pima Indian populations. We here present multipoint linkage analysis of adiponectin levels using a variance components model for 1007 siblings (from 360 nuclear families) of Chinese origin and 352 siblings (from 147 nuclear families) of Japanese origin. We found heritability for adiponectin concentrations was 0.70 for Chinese and 0.48 for Japanese. Autosomal genome scan was performed using microsatellite markers span at an interval of approximately 10 cM. Suggestive linkage of adiponectin, after adjusting for age and sex, was found on chromosome 15 at 39 cM (maximal LOD score = 3.19, P = 6.3 x 10(-5)) for Chinese; and on chromosome 18 at 28 cM (maximal LOD score = 2.40, P = 4.4 x 10(-4)) for Japanese. There were tentative loci of weak linkage on chromosomes 3, 18, and 20 in Japanese. We provide novel loci on chromosomes 15, 18, and 20 and confirm a region on chromosome 3 as reported in Pima Indians, which may influence differentially on circulating adiponectin concentrations in Chinese and Japanese populations. Further fine mapping of these regions will help to identify the gene(s) that might affect adiponectin levels. PMID- 15531542 TI - A familial form of congenital hypopituitarism due to a PROP1 mutation in a large kindred: phenotypic and in vitro functional studies. AB - We report the natural history of a hypopituitarism in a large Tunisian kindred including 29 subjects from the same consanguineous family. The index case was a 9 yr-old girl with severe growth retardation due to complete GH deficiency and partial corticotroph, lactotroph, and thyrotroph deficiencies. Magnetic resonance imaging showed a hyperplastic anterior pituitary. Thirteen of the 28 relatives examined (10 female subjects) had hypopituitarism. In the 14 patients, previously untreated (aged 6-53 yr), height was -5.7 +/- 1.7 sd score, and puberty was spontaneously initiated in only two females. Complete GH deficiency was found in all 12 patients investigated, of whom 11 had thyrotroph and eight of 10 had corticotroph deficiency. A homozygous R73C mutation of PROP1 was present in all 10 patients studied, and a heterozygous mutation was found in six unaffected parents or siblings. In vitro the mutant had 11.5% of the transactivation capacity of the wild type and was unable to bind to a high-affinity DNA sequence. This report showed the deleterious effect of the recessive R73C mutation that affects a hot spot of the PROP1 gene and was associated with severe dwarfism, a lack of spontaneous puberty, and a high incidence of early onset of corticotroph deficiency. PMID- 15531543 TI - Low prevalence of thyrotropin receptor mutations in a large series of subjects with sporadic and familial nonautoimmune subclinical hypothyroidism. AB - Subclinical hypothyroidism of chronic autoimmune thyroiditis must be distinguished from the rare condition of thyroid resistance to TSH in which variable degrees of congenital insensitivity of the thyroid to a biologically active TSH molecule are present. We studied 42 subjects with slight to moderate elevations of circulating TSH and normal free thyroid hormone levels in whom the diagnosis of autoimmune thyroid disease had been excluded using the best of the currently available laboratory and instrumental techniques. In three families (A, B, and C), which included 8 of the 42 cases, other members besides the propositus were found to have isolated hyperthyrotropinemia. The entire coding regions of the TSH receptor (TSHr) gene were sequenced, and TSHr mutations were found in five subjects from families A and B. No mutations were identified in the two members of family C, in one member of family A, and in the 34 remaining cases of isolated hyperthyrotropinemia. A previously described P162A mutation was found in the proband (homozygous state), the son, and the mother of family A (both in the heterozygous state). A new inactivating heterozygous mutation was found in the proband and the mother of family B and consisted of the substitution of a leucine in place of a highly conserved proline at position 252 (L252P) in the extracellular portion of the TSHr. After transfection in COS-7 cells, the mutant L252P displayed a low expression at the cell surface and a reduced response to bovine TSH in terms of cAMP production. A structural defect of the mutant TSHr protein was probably responsible for the poor routing of the receptor to the cell membrane. In conclusion, in two of three families, but in none of 34 sporadic cases of isolated hyperthyrotropinemia, inactivating mutations of the TSHr were identified. The question of whether the latter cases represent subtle forms of autoimmune thyroiditis or might bear as yet unidentified genetic defects remains a matter of future studies. PMID- 15531544 TI - Regulation of angiogenic activity of human endometrial endothelial cells in culture by ovarian steroids. AB - Blood vessel growth and regression in human endometrium are regulated throughout the menstrual cycle. We sought a direct role of ovarian steroids on human endometrial endothelial cell (HEEC) proliferation and vascularization. To investigate the HEEC angiogenicity of sex steroids, we developed a reliable method for the isolation of HEEC, which allowed us to investigate the angiogenic effects of sex steroids using immunohistochemistry, immunocytochemistry, Western blot, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl) 2H-tetrazolium, inner salt proliferation, and vascular tube formation analyses. We were able to obtain 95-99% pure HEEC with our isolation technique. HEEC expressed predominantly estrogen receptor beta, minimally expressed estrogen receptor alpha, and but did not express progesterone (P(4)) receptors A and B in vivo and in vitro. Estradiol (E(2); 10(-10)-10(-8) m) and P(4) (10(-12)-10(-8) m), alone or in combination, induced HEEC proliferation compared with control values after 48 h of treatment (P < 0.05). Furthermore, after 8 d of treatment, there were significantly more angiogenic patterns in E(2) (10(-8) m), P(4) (10( 10) m), and E(2) plus P(4) (10(-8) and 10(-10) m) treatment groups compared with the control group (angiogenic scores, 2.95 +/- 0.16, 3.26 +/- 0.16, 3.06 +/- 0.17, and 1.93 +/- 0.15, respectively; P < 0.01). In conclusion, our results suggest that there are direct effects of E(2) and P(4) on HEEC and provide a new understanding of the physiological role of sex steroids in the regulation of endometrial events such as angiogenesis. PMID- 15531545 TI - Regulation of placental vascular endothelial growth/permeability factor expression and angiogenesis by estrogen during early baboon pregnancy. AB - We have recently shown that there was a developmental increase in placental trophoblast vascular endothelial growth/permeability factor (VEG/PF) expression and vascularization that closely paralleled maternal serum estrogen levels during advancing baboon gestation. The present study determined whether estrogen regulates these important aspects of primate development. VEG/PF mRNA levels were determined by competitive RT-PCR in isolated villous placental cells, and placental vascularization was assessed by image analysis. Placentas were obtained on d 60 of gestation (length of gestation is 184 d) from baboons in which estrogen levels on d 25-59 were increased by daily administration of aromatizable androstenedione or decreased by aromatase inhibitor CGS 20267. Androstenedione treatment increased maternal serum estradiol levels 3-fold (P < 0.01) and placental villous cytotrophoblast VEG/PF mRNA level to a value (mean +/- se, 26,836 +/- 5,625 attomoles/microg total RNA) 2.5-fold greater (P < 0.05) than that in untreated animals (11,645 +/- 1,746 attomoles/microg RNA). In contrast, administration of CGS 20267 decreased serum estradiol (P < 0.01) and placental cytotrophoblast mRNA (2,912 +/- 693 attomoles/microg RNA; P < 0.05) levels by 75%, effects prevented by concomitant administration of CGS 20267 and estradiol. VEG/PF mRNA levels in inner villous cells were unaltered. Coinciding with the increase in placental VEG/PF expression, the percent vascularized area (3.46 +/- 0.23) and vessel density (493 +/- 34 vessels/mm(2)) of the villous placenta in untreated baboons on d 60 were increased (P < 0.01) in baboons in which estrogen levels were elevated by androstenedione treatment (6.54 +/- 0.56 and 743 +/- 27 vessels/mm(2), respectively). It is concluded that estrogen has an important role in stimulating trophoblast VEG/PF expression and consequently villous placental angiogenesis to promote fetal growth and development in early primate pregnancy. PMID- 15531547 TI - Residual activity of mutant androgen receptors explains wolffian duct development in the complete androgen insensitivity syndrome. AB - Development of the Wolffian ducts (WD) into epididymides and vasa deferentia is dependent on testosterone. Patients with the complete androgen insensitivity syndrome (CAIS) are therefore not expected to develop these structures. However, WD derivatives have been described in cases of CAIS. It is thought that these may be remnants. This study assesses the degree of WD development in 33 patients with CAIS and investigates whether this development was androgen dependent. Epididymides and vasa deferentia were identified in 70% of patients with substitution mutations in the androgen receptor ligand-binding domain. They were more developed than epididymides and vasa deferentia from 16- to 20-wk-old male fetuses, suggesting that the WD had been stimulated to grow, rather than failed to regress. Receptors with substitutions in the ligand-binding domain were normally expressed and showed residual response to androgens in transactivation assays. Patients with premature stop codons or frameshift mutations, which prevented androgen receptor expression, or DNA-binding domain mutations that abolished transcriptional activity did not have epididymides or vasa deferentia. We hypothesize that mutant receptors with residual activity in vitro respond to high local testosterone concentrations in vivo, thereby stimulating WD development. The classification of androgen insensitivity in such patients should be considered severe rather than complete. PMID- 15531546 TI - Thyroid fetal male microchimerisms in mothers with thyroid disorders: presence of Y-chromosomal immunofluorescence in thyroid-infiltrating lymphocytes is more prevalent in Hashimoto's thyroiditis and Graves' disease than in follicular adenomas. AB - The presence of fetal cells in a maternal compartment is defined as fetal maternal microchimerism, which has been detected in thyroids of mothers suffering from autoimmunity. We analyzed the immunohistology of paraffin-embedded thyroid specimen taken at surgery from 49 women with Hashimoto's thyroiditis (n = 25), Graves' disease (n = 15), or nodular or diffuse follicular adenomas (n = 9), whose childbirth history was positive for sons. By fluorescence in situ hybridization we screened for X-chromosome- and Y-chromosome-specific staining and compared the finding with human leukocyte antigen (HLA) DQ types of the mothers and, where available, their offspring. In 23 thyroids we found Y chromosome-specific staining, which was more frequent in thyroid autoimmune disease (60% Hashimoto's thyroiditis and 40% Graves' disease) than in follicular adenomas (22.2%). There was no significant difference for HLA DQ alleles among women whose thyroids showed Y-chromosome staining and those without. However, a subgroup of all investigated microchimerism-positive mother-child pairs and women with Hashimoto's thyroiditis and Graves' disease more often had the susceptibility alleles HLA DQA1*0501-DQB1*0201 or DQB1*0301. In conclusion, fetal microchimerism is observed in thyroids of mothers with sons, and this is found more frequently in thyroid autoimmune diseases. PMID- 15531548 TI - Identification of a novel point mutation in the RET gene (Ala883Thr), which is associated with medullary thyroid carcinoma phenotype only in homozygous condition. AB - The RET protooncogene mutations responsible for multiple endocrine neoplasia type 2 are inherited as autosomic dominant traits. We describe here a novel germline homozygous mutation in exon 15 of the RET gene that determines an amino acid substitution (Ala->Thr) at codon 883. The index case was a 51-yr-old patient with an apparently sporadic form of medullary thyroid cancer (MTC). RET gene mutations screening was performed in exons 10, 11, 13, 14, 15, and 16 by automatic sequence analysis. An unexpected homozygous GCT->ACT point mutation was found at codon 883 in exon 15 and confirmed by restriction analysis (Alu I). The presence of the two chromosomes 10 was confirmed by fluorescence in situ hybridization analysis on lymphocytes. As expected on the basis of the homozygosity of the index case, the parents were consanguineous (second-degree cousins). Eight relatives were further investigated: the mother, two sisters, and the son were positive for heterozygous RET mutation. The mother (82 yr old) showed a nodular goiter but was negative both for basal and pentagastrin stimulated calcitonin. The young son (15 yr old) and the two sisters (63 and 58 yr old) did not show any clinical and/or biochemical sign of MTC. One brother (59 yr old) was negative both for RET mutation and clinical/biochemical examination. The other brother, 56 yr of age, was positive for both homozygous RET mutation and serum calcitonin. When operated, the histological examination of the thyroid showed the presence of MTC and C cell hyperplasia. In conclusion, we identified a new germline RET gene mutation during a routine RET gene screening of an apparently sporadic MTC case. This mutation showed a very low transforming activity as demonstrated by the absence of MTC phenotype in heterozygous subjects. The possibility that the homozygous gene carriers were indeed carrying a germline loss of heterozygosity was excluded by fluorescence in situ hybridization analysis for RET gene performed on lymphocytes derived from one homozygous patient. The analysis of several RET polymorphisms also confirmed the presence of two mutated alleles in MTC affected patients and both mutated and wild-type allele in heterozygous subjects. PMID- 15531549 TI - Involvement of membrane-type matrix metalloproteinases (MT-MMPs) in capillary tube formation by human endometrial microvascular endothelial cells: role of MT3 MMP. AB - In the endometrium, angiogenesis is a physiological process, whereas in most adult tissues neovascularization is initiated only during tissue repair or pathological conditions. Pericellular proteolysis plays an important role in angiogenesis being required for endothelial cell migration, invasion, and tube formation. We studied the expression of proteases by human endometrial microvascular endothelial cells (hEMVECs) and their involvement in the formation of capillary tubes and compared these requirements with those of foreskin MVECs (hFMVECs). Inhibition of urokinase and matrix metalloproteinase (MMP) both reduced tube formation in a fibrin or fibrin/collagen matrix. hEMVECs expressed various MMP mRNAs and proteins; in particular MMP-1, MMP-2, and membrane-type (MT)1-, MT3-, and MT4-MMPs. MT3- and MT4-MMP mRNA expressions were significantly higher in hEMVECs than in hFMVECs. Other MT-MMP mRNAs and MMP-9 were hardly detectable. Immunohistochemistry confirmed the presence of MT3-MMP in endothelial cells of endometrial tissue. Overexpression of tissue inhibitor of MMP (TIMP)-1 or TIMP-3 by adenoviral transduction of hEMVECs reduced tube formation to the same extent, whereas only TIMP-3 was able to inhibit tube formation by hFMVECs. Tube formation by hEMVECs was partly inhibited by the presence of anti-MT3-MMP IgG. Thus, in contrast to tube formation by hFMVECs, which largely depends on MT1 MMP, capillary-like tube formation by hEMVECs is, at least in part, regulated by MT3-MMP. PMID- 15531550 TI - Identification and characterization of a novel luciferase-like protein in the human female reproductive tract. AB - A novel cDNA was cloned from human endometrium, matching a human gene with the interim name KIAA1463. An mRNA identified by 5'-rapid amplification of cDNA ends was found to be 3349 nt in length. PCR analysis also identified another transcript of 6626 nt, with an open reading frame encoding a 900 amino acid protein. A fold recognition program identified similarity to firefly luciferase containing an AMP-binding motif; hence, we refer to the predicted protein as the AMP binding/luciferase-like protein (ALLP). ALLP mRNA and protein were expressed throughout the female reproductive tract with the highest levels found in the ovary and uterus. In situ hybridization and immunohistochemistry showed predominant localization of the ALLP mRNA/protein in endometrial glandular epithelium and within the theca and granulosa cells in the ovary. In the endometrium expression of ALLP, mRNA and protein were higher during d 16-21 of the secretory phase of the cycle. Western blot analysis showed decreased expression of ALLP in the postmenopausal endometrium, and hormone replacement therapy increased the expression of ALLP. Endometrial adenocarcinoma cell lines expressed more ALLP, compared with cultured primary endometrial cells or normal endometrial tissue. The ubiquitous expression of ALLP in reproductive and nonreproductive tissues suggests that this protein, which is probably regulated by ovarian steroids, plays an important metabolic role and may be involved in such processes as implantation and tumorigenesis. PMID- 15531551 TI - Novel mutations of the chloride channel Kb gene in two Japanese patients clinically diagnosed as Bartter syndrome with hypocalciuria. AB - Hypokalemic metabolic tubulopathy, such as in Bartter syndrome and Gitelman syndrome, is caused by the dysfunction of renal electrolyte transporters. Despite advances in molecular genetics with regard to hypokalemic metabolic tubulopathy, recent reports have suggested that the phenotype-genotype correlation is still confusing, especially in classic Bartter and Gitelman syndromes. We report here two Japanese patients who suffered from clinically diagnosed classic Bartter syndrome but who presented hypocalciuria. Hypocalciuria is generally believed to be a pathognomonic finding of NCCT malfunction. To better understand the genotype phenotype correlation in these two cases, we screened four renal electrolyte transporter genes [Na-K-2Cl cotransporter (NKCC2), renal outer medullary K channel (ROMK), Cl channel Kb (ClC-Kb), and Na-Cl cotransporter (NCCT)] by the PCR direct sequencing method. We identified three ClC-Kb allelic variants, including two new mutations (L27R and W610X in patient 1 and a G to C substitution of a 3' splice site of intron 2 and W610X in patient 2). We did not find any mutations in the other three genes. Our present data suggest that some ClC-Kb mutations may affect calcium handling in renal tubular cells. PMID- 15531552 TI - Decreased expression of retinoid X receptor isoforms in human thyroid carcinomas. AB - Retinoid X receptors (RXRs) are ligand-inducible transcription factors that belong to the superfamily of nuclear hormone receptors. Because RXRs heterodimerize with thyroid hormone receptor, retinoic acid receptor, vitamin D(3) receptor, and peroxisome proliferator-activated receptor, they play central roles in regulating a number of signaling pathways. To understand the roles of RXRs in human thyroid carcinogenesis, we have investigated the immunohistochemical expression of RXRs in normal and neoplastic thyroid tissues. Whereas nontumorous human thyroid cells exhibited distinct nuclear staining for the RXRs, thyroid carcinomas showed decreased nuclear expression of all three RXR isoforms. In particular, some thyroid carcinoma cells showed intense RXR-alpha cytoplasmic staining accompanied by decreased immunoreactivity in their nuclei. This subcellular localization of RXR-alpha was confirmed by Western blot analysis, which showed both lower nuclear expression levels of RXR-alpha and a cytosolic presence of RXR-related protein in neoplastic regions. We present here, for the first time, the histological distribution of each RXR protein (alpha, beta, and gamma) in human thyroid follicular cells. In addition, we found that the nuclear expression of RXRs was lower in thyroid carcinomas than in normal tissue. The differential expressions of these RXRs in thyroid carcinomas might be implicated in the pathogenesis of thyroid cancers. PMID- 15531553 TI - The codon 620 tryptophan allele of the lymphoid tyrosine phosphatase (LYP) gene is a major determinant of Graves' disease. AB - The lymphoid tyrosine phosphatase (LYP), encoded by the protein tyrosine phosphatase-22 (PTPN22) gene, is a powerful inhibitor of T cell activation. Recently, a single nucleotide polymorphism (SNP), encoding a functional arginine to tryptophan residue change at LYP codon 620 has been shown to be associated with type 1 diabetes and other autoimmune disorders. We have used a PCR restriction fragment (XcmI) assay to examine genotypes at the codon 620 polymorphism in 549 unrelated probands with Graves' disease, 104 unrelated subjects with autoimmune Addison's disease and 429 controls. The T nucleotide at the SNP, encoding the tryptophan 620 residue, was present in 151 of 1098 (13.8%) Graves' disease alleles compared to 67 of 858 (7.8%) control alleles (chi(2) = 17.2, p = 3.4 x 10(-5)' odds ratio = 1.88, 5-95% confidence intervals [CI] 1.39 to 2.55). Similarly, the T nucleotide at the codon 620 SNP was present in 26 of 208 (12.5%) Addison's disease alleles vs 7.8% of controls (chi(2) = 4.63, p = 0.031; odds ratio = 1.69, 5-95% CI 1.04 to 2.73). These data suggest that this LYP polymorphism is a susceptibility allele for Graves' disease with a major effect, and which is likely to have a role in many other autoimmune conditions. PMID- 15531554 TI - Rapid preoperative preparation for severe hyperthyroid Graves' disease. PMID- 15531555 TI - Relationship of sex hormones to bone geometric properties and mineral density in early pubertal girls: use of correlation analyses. PMID- 15531556 TI - Evidence for the use of igf-I as a predictor of mortality in acromegaly is lacking. PMID- 15531557 TI - The relationship of fasting serum radioimmune insulin levels to incident coronary heart disease in an insulin-treated diabetic cohort. PMID- 15531558 TI - Afferent mechanisms underlying stimulation modality-related modulation of acupuncture-related cardiovascular responses. AB - Despite the use of acupuncture to treat a number of heart diseases, little is known about the mechanisms that underlie its actions. Therefore, we examined the influence of acupuncture on sympathoexcitatory cardiovascular responses to gastric distension in anesthetized Sprague-Dawley rats. Thirty minutes of low current, low-frequency, (0.3-0.5 mA, 2 Hz) electroacupuncture (EA), at P 5-6, S 36-37, and H 6-7 overlying the median, deep peroneal, and ulnar nerves significantly decreased reflex pressor responses by 40, 39, and 44%, respectively. In contrast, sham acupuncture involving needle insertion without stimulation at P 5-6 or 30 min of EA at LI 6-7 acupoints overlying the superficial radial nerve did not attenuate the reflex. Similarly, EA at P 5-6 using 40- or 100-Hz stimulation frequencies did not inhibit the reflex. Compared with EA at P 5-6, EA at two sets of acupoints, including P 5-6 and S 36-37, did not lead to larger inhibition of the reflex. Two minutes of manual acupuncture (MA; 2 Hz) at P 5-6 every 10 min for 30 min inhibited the reflex cardiovascular pressor response by 33%, a value not significantly different from 2-Hz EA at P 5 6. Single-unit afferent activity was not different between electrical stimulation (ES) and manual stimulation. However, 2-Hz ES activated more somatic afferents than 10- or 20-Hz ES. These data suggest that, although the location of acupoint stimulation and the frequency of stimulation determine the extent of influence of EA, there is little difference between low-frequency EA and MA at P 5-6. Furthermore, simultaneous stimulation using two acupoints that independently exert strong effects did not lead to an additive or a facilitative interaction. The similarity of the responses to EA and MA and the lack of cardiovascular response to high-frequency EA appear to be largely a function of somatic afferent responses. PMID- 15531559 TI - Monocarboxylate transporters, blood lactate removal after supramaximal exercise, and fatigue indexes in humans. AB - The present study investigated whether muscular monocarboxylate transporter (MCT) 1 and 4 contents are related to the blood lactate removal after supramaximal exercise, fatigue indexes measured during different supramaximal exercises, and muscle oxidative parameters in 15 humans with different training status. Lactate recovery curves were obtained after a 1-min all-out exercise. A biexponential time function was then used to determine the velocity constant of the slow phase (gamma(2)), which denoted the blood lactate removal ability. Fatigue indexes were calculated during 1-min all-out (FI(AO)) and repeated 10-s (FI(Sprint)) cycling sprints. Biopsies were taken from the vastus lateralis muscle. MCT1 and MCT4 contents were quantified by Western blots, and maximal muscle oxidative capacity (V(max)) was evaluated with pyruvate + malate and glutamate + malate as substrates. The results showed that the blood lactate removal ability (i.e., gamma(2)) after a 1-min all-out test was significantly related to MCT1 content (r = 0.70, P < 0.01) but not to MCT4 (r = 0.50, P > 0.05). However, greater MCT1 and MCT4 contents were negatively related with a reduction of blood lactate concentration at the end of 1-min all-out exercise (r = -0.56, and r = -0.61, P < 0.05, respectively). Among skeletal muscle oxidative indexes, we only found a relationship between MCT1 and glutamate + malate V(max) (r = 0.63, P < 0.05). Furthermore, MCT1 content, but not MCT4, was inversely related to FI(AO) (r = 0.54, P < 0.05) and FI(Sprint) (r = -0.58, P < 0.05). We concluded that skeletal muscle MCT1 expression was associated with the velocity constant of net blood lactate removal after a 1-min all-out test and with the fatigue indexes. It is proposed that MCT1 expression may be important for blood lactate removal after supramaximal exercise based on the existence of lactate shuttles and, in turn, in favor of a better tolerance to muscle fatigue. PMID- 15531560 TI - Differences in time-dependent hypoxic phrenic responses among inbred rat strains. AB - Hypoxic ventilatory responses differ between rodent strains, suggesting a genetic contribution to interindividual variability. However, hypoxic ventilatory responses consist of multiple time-dependent mechanisms that can be observed in different respiratory motor outputs. We hypothesized that strain differences would exist in discrete time-dependent mechanisms of the hypoxic response and, furthermore, that there may be differences between hypoglossal and phrenic nerve responses to hypoxia. Hypoglossal and phrenic nerve responses were assessed during and after a 5-min hypoxic episode in anesthetized, vagotomized, and ventilated rats from four inbred strains: Brown Norway (BN), Fischer 344 (FS), Lewis (LW), and Piebald-viral-Glaxo (PVG). During baseline, burst frequency was higher in PVG than LW rats (P < 0.05), phrenic burst amplitude was higher in PVG vs. other strains (P < 0.05), and hypoglossal burst amplitude was higher in PVG and BN vs. FS and LW (P < 0.05). During hypoxia, burst frequency did not change in BN or LW rats, but it increased in PVG and FS rats. The phrenic amplitude response was smallest in PVG vs. other strains (P < 0.05), and the hypoglossal response was similar among strains. Short-term potentiation posthypoxia was slowest in FS and fastest in LW rats (P < 0.05). Posthypoxia frequency decline was absent in PVG, but it was observed in all other strains. Augmented breaths were observed during hypoxia in FS rats only. Thus genetic differences exist in the time domains of the hypoxic response, and these are differentially expressed in hypoglossal and phrenic nerves. Furthermore, genetic diversity observed in hypoxic ventilatory responses in unanesthetized rats may arise from multiple neural mechanisms. PMID- 15531561 TI - Hypoxia reveals posterior thalamic, cerebellar, midbrain, and limbic deficits in congenital central hypoventilation syndrome. AB - Congenital central hypoventilation syndrome (CCHS) patients show deficient respiratory and cardiac responses to hypoxia and hypercapnia, despite apparently intact arousal responses to hypercapnia and adequate respiratory motor mechanisms, thus providing a model to evaluate functioning of particular brain mechanisms underlying breathing. We used functional magnetic resonance imaging to assess blood oxygen level-dependent signals, corrected for global signal changes, and evaluated them with cluster and volume-of-interest procedures, during a baseline and 2-min hypoxic (15% O(2), 85% N(2)) challenge in 14 CCHS and 14 age- and gender-matched control subjects. Hypoxia elicited significant (P < 0.05) differences in magnitude and timing of responses between groups in cerebellar cortex and deep nuclei, posterior thalamic structures, limbic areas (including the insula, amygdala, ventral anterior thalamus, and right hippocampus), dorsal and ventral midbrain, caudate, claustrum, and putamen. Deficient responses to hypoxia included no, or late, changes in CCHS patients with declining signals in control subjects, a falling signal in CCHS patients with no change in controls, or absent early transient responses in CCHS. Hypoxia resulted in signal declines but no group differences in hypothalamic and dorsal medullary areas, the latter being a target for PHOX2B, mutations of which occur in the syndrome. The findings extend previously identified posterior thalamic, midbrain, and cerebellar roles for normal mediation of hypoxia found in animal fetal and adult preparations and suggest significant participation of limbic structures in responding to hypoxic challenges, which likely include cardiovascular and air-hunger components. Failing structures in CCHS include areas additional to those associated with PHOX2B expression and chemoreceptor sites. PMID- 15531562 TI - Effects of a long-term spaceflight on immunoglobulin heavy chains of the urodele amphibian Pleurodeles waltl. AB - A variety of immune parameters are modified during and after a spaceflight. The effects of spaceflights on cellular immunity are well documented; however, little is known about the effects of these flights on humoral immunity. During the Genesis space experiment, two adult Pleurodeles waltl (urodele amphibian) stayed 5 mo onboard Mir and were subjected to oral immunization. Animals were killed 10 days after their return to earth. IgM and IgY heavy-chain transcripts in their spleens were quantified by Northern blotting. The use of the different VH families (coding for antibody heavy-chain variable domains) in IgM heavy chain transcripts was also analyzed. Results were compared with those obtained with ground control animals and animals reared in classical conditions in our animal facilities. We observed that, 10 days after the return on earth, the level of IgM heavy-chain transcription was normal but the level of IgY heavy-chain transcription was at least three times higher than in control animals. We also observed that the use of the different VH families in IgM heavy-chain transcripts was modified by the flight. These data suggest that the spaceflight affected the antibody response against the antigens contained in the food. PMID- 15531563 TI - Decreased contraction-stimulated glucose transport in isolated epitrochlearis muscles of pregnant rats. AB - Late pregnancy is characterized by insulin resistance for glucose transport in skeletal muscle. The main purpose of this study was to investigate the effect of late pregnancy on contraction-stimulated glucose transport in isolated rat skeletal muscle after in vitro electrical stimulation. Isolated epitrochlearis muscles of 19-day pregnant and aged-matched nonpregnant control rats were studied. One muscle from each rat was stimulated to contract, and the contralateral muscle served as a resting control. Tension developed during contractile activity, 3-O-methylglucose (3-MG) transport rate, and glycogen concentration were determined. Epitrochlearis muscles from other rats were used to measure insulin-stimulated 3-MG transport. There was no detectable difference between the nonpregnant and pregnant groups for contractile performance (peak tension, total tension, or fatigue). Pregnancy was not associated with significant changes in muscle glycogen concentration (resting or after contractile activity) or the contraction-stimulated decrement in glycogen concentration. For muscles from pregnant vs. nonpregnant groups, there was a 22% reduction (P < or = 0.05) in contraction-stimulated glucose transport, a 28% decrease (P < or = 0.05) in insulin-stimulated glucose transport, and unchanged basal glucose transport. In conclusion, isolated epitrochlearis muscles from pregnant vs. nonpregnant rats had a relative decrement in contraction-stimulated glucose transport that was similar to the relative decline in insulin-stimulated glucose transport. The decrement in contraction-stimulated glucose transport was not attributable to pregnancy-related changes in tension development or glycogen levels. The similar relative decline in insulin- and contraction-stimulated glucose transport raises the possibility that pregnancy impairs a distal process that is common to mechanisms whereby each stimulus activates glucose transport. PMID- 15531564 TI - Circumferential vascular deformation after stent implantation alters wall shear stress evaluated with time-dependent 3D computational fluid dynamics models. AB - The success of vascular stents in the restoration of blood flow is limited by restenosis. Recent data generated from computational fluid dynamics (CFD) models suggest that stent geometry may cause local alterations in wall shear stress (WSS) that have been associated with neointimal hyperplasia and subsequent restenosis. However, previous CFD studies have ignored histological evidence of vascular straightening between circumferential stent struts. We tested the hypothesis that consideration of stent-induced vascular deformation may more accurately predict alterations in indexes of WSS that may subsequently account for histological findings after stenting. We further tested the hypothesis that the severity of these alterations in WSS varies with the degree of vascular deformation after implantation. Steady-state and time-dependent simulations of three-dimensional CFD arteries based on canine coronary artery measurements of diameter and blood flow were conducted, and WSS and WSS gradients were calculated. Circumferential straightening introduced areas of high WSS between stent struts that were absent in stented vessels of circular cross section. The area of vessel exposed to low WSS was dependent on the degree of circumferential vascular deformation and axial location within the stent. Stents with four vs. eight struts increased the intrastrut area of low WSS in vessels, regardless of cross-sectional geometry. Elevated WSS gradients were also observed between struts in vessels with polygonal cross sections. The results obtained using three dimensional CFD models suggest that changes in vascular geometry after stent implantation are important determinants of WSS distributions that may be associated with subsequent neointimal hyperplasia. PMID- 15531565 TI - Cardiovagal regulation during combined hypoxic and orthostatic stress: fainters vs. nonfainters. AB - We tested the hypothesis that individual differences in the effect of acute hypoxia on the cardiovagal arterial baroreflex would determine individual susceptibility to hypoxic syncope. In 16 healthy, nonsmoking, normotensive subjects (8 women, 8 men, age 20-33 yr), we assessed orthostatic tolerance with a 20-min 60 degrees head-upright tilt during both normoxia and hypoxia (breathing 12% O(2)). On a separate occasion, we assessed baroreflex control of heart rate (cardiovagal baroreflex gain) using the modified Oxford technique during both normoxia and hypoxia. When subjects were tilted under hypoxic conditions, 5 of the 16 developed presyncopal signs or symptoms, and the 20-min tilt had to be terminated. These "fainters" had comparable cardiovagal baroreflex gain to "nonfainters" under both normoxic and hypoxic conditions (normoxia, fainters: 1.2 +/- 0.2, nonfainters: -1.0 +/- 0.2 beats.min(-1).mmHg(-1), P = 0.252; hypoxia, fainters: -1.3 +/- 0.2, nonfainters: -1.0 +/- 0.1 beats.min(-1).mmHg( 1), P = 0.208). Furthermore, hypoxia did not alter cardiovagal baroreflex gain in either group (both P > 0.8). It appears from these observations that hypoxic syncope results from the superimposed vasodilator effects of hypoxia on the cardiovascular system and not from a hypoxia-induced maladjustment in baroreflex control of heart rate. PMID- 15531566 TI - Nonsymmetrical double logistic analysis of ambulatory blood pressure recordings. AB - We developed an asymmetric double logistic curve-fitting procedure for circadian analysis that can determine the rate of change in variables during the day-to night separately from the night-to-day transition for use in animal studies. We now have applied this procedure to 24-h systolic (SAP) and diastolic arterial pressure (DAP) and heart rate ambulatory recordings from 302 patients. In 292 cases, all parameters showed a pattern of higher day and lower night values. In men there was a similar rate of transition between day and night or from night to day for both SAP and DAP that lasted 3-4 h, indicating a symmetrical diurnal pattern. By contrast, women showed a faster rate of decrease in mean arterial pressure in the evening compared with men (P < 0.05) and therefore showed an asymmetric diurnal SAP pattern. For both men and women, there was a markedly greater rate of morning increase in heart rate compared with the rate of evening decrease (2.2- and 1.9-fold, respectively, P < 0.001). The logistic method provided a better fit than the square-wave or the cosinor method (P < 0.001) and more appropriately detected nondippers. We conclude that analysis of ambulatory recordings by a new logistic curve-fitting method reveals more rapid reductions in evening SAP in women than men but both have two- to threefold more rapid morning rates of tachycardia. The ability of the double logistic method to determine the diurnal blood pressure rates of change independently is key to determining new markers for cardiovascular risk. PMID- 15531567 TI - 20-HETE-mediated vasoconstriction by hemoglobin-O2 carrier in Sprague-Dawley but not Wistar rats. AB - Hypothetically either decreased nitric oxide (NO) or increased O(2) could initiate 20-HETE-mediated vasoconstriction associated with hemoglobin-based blood substitutes (HBOC). To test this hypothesis, we infused Tm-Hb, an HBOC with low O(2) affinity, into isoflurane-anesthetized Wistar (W) and Sprague-Dawley (SD) rats after exchanging 20% of their blood with Ringer lactate. For comparison we infused an equal amount of BSA or BSA with N(G)-nitro-L-arginine methyl ester (BSA + NAME). Tm-Hb increased blood pressure (BP) and renal vascular resistance (RVR) equally in W and SD rats. Renal blood flow (RBF; Doppler ultrasound) decreased. BSA decreased RVR and raised glomerular filtration rate. BSA + NAME raised BP, RVR, and GFR. HET0016, an inhibitor of 20-HETE production, blunted BP and RVR responses to Tm-Hb and BSA+NAME in SD but not W rats. Arterial O(2) content with BSA was lower than with Tm-Hb but O(2) delivery was 60% higher with BSA because of higher RBF. BSA raised Po(2) (Oxylite) in cortex and medulla and reduced RVR. Tm-Hb decreased Po(2) and increased RVR. Switching rats from breathing air to 100% O(2) raised intrarenal Po(2) two- to threefold and increased BP and RVR. HET0016 did not alter hyperoxic responses. In conclusion, 20-HETE contributes to vasoconstriction by Tm-Hb in SD but not in W rats, and increased 20-HETE activity results primarily from decreased NO. PMID- 15531568 TI - Endurance exercise training increases insulin responsiveness in isolated adipocytes through IRS/PI3-kinase/Akt pathway. AB - Endurance exercise training promotes important metabolic adaptations, and the adipose tissue is particularly affected. The aim of this study was to investigate how endurance exercise training modulates some aspects of insulin action in isolated adipocytes and in intact adipose tissue. Male Wistar rats were submitted to daily treadmill running (1 h/day) for 7 wk. Sedentary age-matched rats were used as controls. Final body weight, body weight gain, and epididymal fat pad weight did not show any statistical differences between groups. Adipocytes from trained rats were smaller than those from sedentary rats (205 +/- 16.8 vs. 286 +/ 26.4 pl; P < 0.05). Trained rats showed decreased plasma glucose (4.9 +/- 0.13 vs. 5.3 +/- 0.07 mM; P < 0.05) and insulin levels (0.24 +/- 0.012 vs. 0.41 +/- 0.049 mM; P < 0.05) and increased insulin-stimulated glucose uptake (23.1 +/- 3.1 vs. 12.1 +/- 2.9 pmol/cm(2); P < 0.05) compared with sedentary rats. The number of insulin receptors and the insulin-induced tyrosine phosphorylation of insulin receptor-beta subunit did not change between groups. Insulin-induced tyrosine phosphorylation insulin receptor substrates (IRS)-1 and -2 increased significantly (1.57- and 2.38-fold, respectively) in trained rats. Insulin induced IRS-1/phosphatidylinositol 3 (PI3)-kinase (but not IRS-2/PI3-kinase) association and serine Akt phosphorylation also increased (2.06- and 3.15-fold, respectively) after training. The protein content of insulin receptor-beta subunit, IRS-1 and -2, did not differ between groups. Taken together, these data support the hypothesis that the increased adipocyte responsiveness to insulin observed after endurance exercise training is modulated by IRS/PI3-kinase/Akt pathway. PMID- 15531569 TI - Shear stress induces eNOS mRNA expression and improves endothelium-dependent dilation in senescent soleus muscle feed arteries. AB - We tested the hypothesis that increased intraluminal shear stress induces endothelial nitric oxide (NO) synthase (eNOS) mRNA expression and improves endothelium-dependent dilation in senescent soleus muscle feed arteries (SFA) by increasing NO production. SFA were isolated from young (4 mo) and old (24 mo) male Fischer 344 rats and cannulated with two resistance-matched glass micropipettes. SFA were exposed to no flow (NF), low flow (LF), intermediate flow (IF), or high flow (HF) for 4 h. Mean intraluminal shear stress ranged from 0 to 82 dyn/cm(2). At the end of the 4-h treatment period, eNOS mRNA expression was assessed in each SFA. eNOS mRNA expression was significantly lower in old NF SFA than in young NF SFA. In old SFA, eNOS mRNA expression was induced by IF (+154%) and HF (+136%), resulting in a level of expression that was not different from that of young SFA. In a separate series of experiments, SFA were pretreated with NF or HF for 4 h, and endothelial function was assessed by examining vasodilator responses to ACh. ACh-induced dilation was less in old NF SFA than young NF SFA. Pretreatment with HF improved ACh-induced dilation in old SFA such that the response was similar to that of young SFA. In the presence of N(omega)-nitro-L arginine to inhibit NOS, ACh-induced dilation was inhibited in old HF SFA such that the response was no longer greater than that of old NF SFA. These results indicate that increased intraluminal shear stress induces eNOS mRNA expression and improves endothelium-dependent dilation in senescent SFA by increasing NO production. PMID- 15531570 TI - On the terminology for describing the length-force relationship and its changes in airway smooth muscle. AB - The observation that the length-force relationship in airway smooth muscle can be shifted along the length axis by accommodating the muscle at different lengths has stimulated great interest. In light of the recent understanding of the dynamic nature of length-force relationship, many of our concepts regarding smooth muscle mechanical properties, including the notion that the muscle possesses a unique optimal length that correlates to maximal force generation, are likely to be incorrect. To facilitate accurate and efficient communication among scientists interested in the function of airway smooth muscle, a revised and collectively accepted nomenclature describing the adaptive and dynamic nature of the length-force relationship will be invaluable. Setting aside the issue of underlying mechanism, the purpose of this article is to define terminology that will aid investigators in describing observed phenomena. In particular, we recommend that the term "optimal length" (or any other term implying a unique length that correlates with maximal force generation) for airway smooth muscle be avoided. Instead, the in situ length or an arbitrary but clearly defined reference length should be used. We propose the usage of "length adaptation" to describe the phenomenon whereby the length-force curve of a muscle shifts along the length axis due to accommodation of the muscle at different lengths. We also discuss frequently used terms that do not have commonly accepted definitions that should be used cautiously. PMID- 15531571 TI - Reversal of muscle fatigue during 16 h of heavy intermittent cycle exercise. AB - This study examined the effects of extended sessions of heavy intermittent exercise on quadriceps muscle fatigue and weakness. Twelve untrained volunteers (10 men and 2 women), with a peak oxygen consumption of 44.3 +/- 2.3 ml.kg( 1).min(-1), exercised at approximately 91% peak oxygen consumption for 6 min once per hour for 16 h. Muscle isometric properties assessed before and after selected repetitions (R1, R2, R4, R7, R12, and R15) were used to quantitate fatigue (before vs. after repetitions) and weakness (before vs. before repetitions). Muscle fatigue at R1 was indicated by reductions (P < 0.05) in peak twitch force (135 +/- 13 vs. 106 +/- 11 N) and by a reduction (P < 0.05) in the force frequency response, which ranged between approximately 53% at 10 Hz (113 +/- 12 vs. 52.6 +/- 7.4 N) and approximately 17% at 50 Hz (324 +/- 27 vs. 270 +/- 30 N). No recovery of force, regardless of stimulation frequency, was observed during the 54 min between R1 and R2. At R2 and for all subsequent repetitions, no reduction in force, regardless of stimulation frequency, was generally found after the exercise. The only exception was for R2, where, at 20 Hz, force was reduced (P < 0.05) by 18%. At R15, force before repetitions for high frequencies (i.e., 100 Hz) returned to R1 (333 +/- 29 vs. 324 +/- 27 N), whereas force at low frequency (i.e., 10 Hz) was only partially (P < 0.05) recovered (113 +/- 12 vs. 70 +/- 6.6 N). It is concluded that multiple sessions of heavy exercise can reverse the fatigue noted early and reduce or eliminate weakness depending on the frequency of stimulation. PMID- 15531572 TI - Acute ventilator-induced vascular permeability and cytokine responses in isolated and in situ mouse lungs. AB - To determine the influence of experimental model and strain differences on the relationship of vascular permeability to inflammatory cytokine production after high peak inflation pressure (PIP) ventilation, we used isolated perfused mouse lung and intact mouse preparations of Balb/c and B6/129 mice ventilated at high and low PIP. Filtration coefficients in isolated lungs and bronchoalveolar lavage (BAL) albumin in intact mice increased within 20-30 min after initiation of high PIP in isolated Balb/c lungs and intact Balb/c, B6/129 wild-type, and p55 and p75 tumor necrosis factor (TNF) dual-receptor null mice. In contrast, the cytokine response was delayed and variable compared with the permeability response. In isolated Balb/c lungs ventilated with 25-27 cmH(2)O PIP, TNF-alpha, interleukin (IL)-1 beta, IL-1 alpha, macrophage inflammatory protein (MIP)-2, and IL-6 concentrations in perfusate were markedly increased in perfusate at 2 and 4 h, but only MIP-2 was detectable in intact Balb/c mice using the same PIP. In intact wild-type and TNF dual-receptor null mice with ventilation at 45 cmH(2)O PIP, the MIP-2 and IL-6 levels in BAL were significantly increased after 2 h in both groups, but there were no differences between groups in the BAL albumin and cytokine concentrations or in lung wet-to-dry weight ratios. TNF-alpha was not be detected in BAL fluids in any group of intact mice. These results suggest that the alveolar hyperpermeability induced by high PIP ventilation occurs very rapidly and is initially independent of TNF-alpha participation and unlikely to depend on MIP-2 or IL-6. PMID- 15531573 TI - Association of interleukin-15 protein and interleukin-15 receptor genetic variation with resistance exercise training responses. AB - Interleukin-15 (IL-15) is an anabolic cytokine that is produced in skeletal muscle and directly affects muscle anabolism in animal and in vitro models. The contribution of IL-15 variability in muscle responses to 10 wk of resistance exercise training in young men and women was examined by measuring acute and chronic changes in IL-15 protein in plasma and characterizing genetic variation in the IL-15 receptor-alpha gene (IL15RA). Participants trained 3 days a week at 75% of one repetition maximum, performing three sets (6-10 repetitions) of 13 resistance exercises. Plasma IL-15 protein was significantly increased (P < 0.05) immediately after acute resistance exercise but did not change with training and was not associated with variability in muscle responses with training. A single nucleotide polymorphism in exon 7 of IL15RA was strongly associated with muscle hypertrophy and accounted for 7.1% of the variation in regression modeling. A polymorphism in exon 4 was also independently associated with muscle hypertrophy and accounted for an additional 3.5% of the variation in hypertrophy. These results suggest that IL-15 is an important mediator of muscle mass response to resistance exercise training in humans and that genetic variation in IL15RA accounts for a significant proportion of the variability in this response. PMID- 15531574 TI - Timing of cortical excitability changes during the reaction time of movements superimposed on tonic motor activity. AB - Seated subjects were instructed to react to an auditory cue by simultaneously contracting the tibialis anterior (TA) muscle of each ankle isometrically. Focal transcranial magnetic stimulation of the leg area of the motor cortex (MCx) was used to determine the time course of changes in motor-evoked potential amplitude (MEP) during the reaction time (RT). In one condition the voluntary contraction was superimposed on tonic EMG activity maintained at 10% of maximal voluntary contraction. In the other condition the voluntary contraction was made starting from rest. MEPs in the TA contralateral to the stimulation coil were evoked at various times during the RT in each condition. These were compared to the control MEPs evoked during tonic voluntary activity or with the subject at rest. The RT was measured trial by trial from the EMG activity of the TA ipsilateral to the magnetic stimulus, taking into account the nearly constant time difference between the two sides. The MEPs became far greater than control MEPs during the RT (mean = 332%, SD = 44 %, of control MEPs, P < 0.001) without any measurable change in the background level of EMG activity. The onset of this facilitation occurred on average 12.80 ms (SD = 7.55 ms) before the RT. There was no difference in the onset of facilitation between the two conditions. Because MEPs were facilitated without a change in the background EMG activity, it is concluded that this facilitation is specifically due to an increase of MCx excitability just before voluntary muscle activation. This conclusion is further reinforced by the observation that MEPs evoked by near-threshold anodal stimuli to the MCx were not facilitated during the RT, in contrast to those evoked by near-threshold transcranial magnetic stimulation. However, several observations in the present and previous studies indicate that MEP amplitude may be more sensitive to alpha motoneuron activity than to motor cortical neuron activity, an idea that has important methodological implications. PMID- 15531575 TI - Cellular and molecular mechanisms involved in branching morphogenesis of the Drosophila tracheal system. AB - Recent comparative studies have shown that, in many instances, the genetic network underlying the development of distinct organ systems is similar in invertebrate and vertebrate organisms. Genetically well-characterized, simple invertebrate model systems, such as Caenorhabditis elegans and Drosophila melanogaster, can thus provide useful insight for understanding more complex organ systems in vertebrates. Here, we summarize recent progress in the genetic analysis of tracheal development in Drosophila and compare the results to studies aimed at a better understanding of lung development in mouse and man. Clearly, both striking similarities and important differences are apparent, but it might still be too early to conclude whether the former or the latter prevail. PMID- 15531577 TI - Comment on "Wall stress misrepresents afterload in children and young adults with abnormal left ventricular geometry". PMID- 15531578 TI - The ETS domain transcription factor Elk-1 regulates the expression of its partner protein, SRF. AB - The ternary complex factors (TCF) are a subfamily of ETS domain transcription factors that bind and activate serum response elements (SREs) in the promoters of target genes in a ternary complex with a second transcription factor, serum response factor (SRF). Here, we have identified the SRF gene as a target for the TCFs, thereby providing a positive feedback loop whereby TCF activation leads to the enhancement of the expression of its partner protein SRF. The binding of the TCF Elk-1 to the SRF promoter and subsequent regulation of SRF expression occurs in a ternary complex-dependent manner. Our data therefore reveal that SRF is an important target for the ERK and Rho signaling pathways that converge on a ternary TCF-SRF complex at the SRE on the SRF promoter. PMID- 15531579 TI - Mimosine attenuates serine hydroxymethyltransferase transcription by chelating zinc. Implications for inhibition of DNA replication. AB - L-mimosine is a naturally occurring plant amino acid and iron chelator that arrests the cell cycle in the late G(1) phase, although its mechanism of action is not known. Some studies indicate that mimosine prevents the initiation of DNA replication, whereas other studies indicate that mimosine disrupts elongation of the replication fork by impairing deoxyribonucleotide synthesis by inhibiting the activity of the iron-dependent enzyme ribonucleotide reductase and the transcription of the cytoplasmic serine hydroxymethyltransferase gene (SHMT1). In this study, the mechanism for mimosine-induced inhibition of SHMT1 transcription was elucidated. A mimosine-responsive transcriptional element was localized within the first 50 base pairs of the human SHMT1 promoter by deletion analyses and gel mobility shift assays. The 50-base-pair sequence contains a consensus zinc-sensing metal regulatory element (MRE) at position -44 to -38, and mutation of the MRE attenuated mimosine-induced transcription repression. Mimosine treatment eliminated MRE- and Sp1-binding activity in nuclear extracts from MCF-7 cells but not in nuclear extracts from a mimosine-resistant cell line, MCF-7/2a. MCF-7 cells cultured in zinc-depleted medium for more than 16 days were viable and lacked cytoplasmic serine hydroxymethyltransferase protein, confirming that mimosine inhibits SHMT1 transcription by chelating zinc. The disruption of DNA protein interactions by zinc chelation provides a general mechanism for the inhibitory effects of mimosine on nuclear processes, including replication and transcription. Furthermore, this study establishes that SHMT1 is a zinc-inducible gene, which provides the first mechanism for the regulation of folate-mediated one-carbon metabolism by zinc. PMID- 15531581 TI - Design of soluble recombinant T cell receptors for antigen targeting and T cell inhibition. AB - The use of recombinant T cell receptors (TCRs) to target therapeutic interventions has been hindered by the naturally low affinity of TCR interactions with peptide major histocompatibility complex ligands. Here, we use multimeric forms of soluble heterodimeric alphabeta TCRs for specific detection of target cells pulsed with cognate peptide, discrimination of quantitative changes in antigen display at the cell surface, identification of virus-infected cells, inhibition of antigen-specific cytotoxic T lymphocyte activation, and identification of cross-reactive peptides. Notably, the A6 TCR specific for the immunodominant HLA A2-restricted human T cell leukemia virus type 1 Tax(11-19) epitope bound to HLA A2-HuD(87-95) (K(D) 120 microm by surface plasmon resonance), an epitope implicated as a causal antigen in the paraneoplastic neurological degenerative disorder anti-Hu syndrome. A mutant A6 TCR that exhibited dramatically increased affinity for cognate antigen (K(D) 2.5 nm) without enhanced cross-reactivity was generated; this TCR demonstrated potent biological activity even as a monomeric molecule. These data provide insights into TCR repertoire selection and delineate a framework for the selective modification of TCRs in vitro that could enable specific therapeutic intervention in vivo. PMID- 15531580 TI - Akt2 phosphorylates ezrin to trigger NHE3 translocation and activation. AB - Initiation of Na(+)-glucose cotransport in intestinal absorptive epithelia causes NHE3 to be translocated to the apical plasma membrane, leading to cytoplasmic alkalinization. We reported recently that this NHE3 translocation requires ezrin phosphorylation. However, the kinase that phosphorylates ezrin in this process has not been identified. Because Akt has also been implicated in NHE3 translocation, we investigated the hypothesis that Akt phosphorylates ezrin. After initiation of Na(+)-glucose cotransport, Akt is activated with kinetics that parallel those of ezrin phosphorylation. Inhibition of p38 MAP kinase, which blocks ezrin phosphorylation, also prevents Akt activation. Purified Akt directly phosphorylates recombinant ezrin at threonine 567 in vitro in an ATP-dependent manner. This in vitro phosphorylation can be prevented by Akt inhibitors. In intact cells, inhibition of either phosphoinositide 3-kinase, an upstream regulator of Akt, or inhibition of Akt itself using inhibitors validated in vitro prevents ezrin phosphorylation after initiation of Na(+)-glucose cotransport. Specific small interfering RNA knockdown of Akt2 prevented ezrin phosphorylation in intact cells. Pharmacological Akt inhibition or Akt2 knockdown also prevented NHE3 translocation and activation after initiation of Na(+)-glucose cotransport, confirming the functional role of Akt2. These studies therefore identify Akt2 as a critical kinase that regulates ezrin phosphorylation and activation. This Akt2 dependent ezrin phosphorylation leads to NHE3 translocation and activation. PMID- 15531582 TI - The pathway for the production of inositol hexakisphosphate in human cells. AB - The yeast and Drosophila pathways leading to the production of inositol hexakisphosphate (InsP(6)) have been elucidated recently. The in vivo pathway in humans has been assumed to be similar. Here we show that overexpression of Ins(1,3,4)P(3) 5/6-kinase in human cell lines results in an increase of inositol tetrakisphosphate (InsP(4)) isomers, inositol pentakisphosphate (InsP(5)) and InsP(6), whereas its depletion by RNA interference decreases the amounts of these inositol phosphates. Expression of Ins(1,3,4,6)P(4) 5-kinase does not increase the amount of InsP(5) and InsP(6), although its depletion does block InsP(5) and InsP(6) production, showing that it is necessary for production of InsP(5) and InsP(6). Expression of Ins(1,3,4,5,6)P(5) 2-kinase increases the amount of InsP(6) by depleting the InsP(5) in the cell, and depletion of 2-kinase decreases the amount of InsP(6) and causes an increase in InsP(5). These results are consistent with a pathway that produces InsP(6) through the sequential action of Ins(1,3,4)P(3) 5/6-kinase, Ins(1,3,4,6)P(4) 5-kinase, and Ins(1,3,4,5,6)P5 2 kinase to convert Ins(1,3,4)P(3) to InsP(6). Furthermore, the evidence implicates 5/6-kinase as the rate-limiting enzyme in this pathway. PMID- 15531583 TI - Myeloperoxidase potentiates nitric oxide-mediated nitrosation. AB - Nitrosation is an important reaction elicited by nitric oxide (NO). To better understand how nitrosation occurs in biological systems, we assessed the effect of myeloperoxidase (MPO), a mediator of inflammation, on nitrosation observed during NO autoxidation. Nitrosation of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ; 10 mum) to 2-nitrosoamino-3-methylimidazo[4,5-f]quinoline (N-NO-IQ) was monitored by HPLC. Using the NO donor spermine NONOate at pH 7.4, MPO potentiated N-NO-IQ formation. The minimum effective quantity of necessary components was 8.5 nm MPO, 0.25 mum H(2)O(2)/min, and 0.024 mum NO/min. Autoxidation was only detected at >/=1.2 mum NO/min. MPO potentiation was not affected by a 40-fold excess flux of H(2)O(2) over NO or less than a 2.4-fold excess flux of NO over H(2)O(2). Potentiation was due to an 8.8-fold increased affinity of MPO-derived nitrosating species for IQ. Autoxidation was inhibited by azide, suggesting involvement of the nitrosonium ion, NO(+). MPO potentiation was inhibited by NADH, but not azide, suggesting oxidative nitrosylation with NO(2)(.) or an NO(2)(.)-like species. MPO nonnitrosative oxidation of IQ with 0.3 mm NO(2)(-) at pH 5.5 was inhibited by azide, but not NADH, demonstrating differences between MPO oxidation of IQ with NO compared with NO(2)(-). Using phorbol ester stimulated human neutrophils, N-NO-IQ formation was increased with superoxide dismutase and inhibited by catalase and NADH, but not NaN(3). This is consistent with nitrosation potentiation by MPO, not peroxynitrite. Increased N-NO-IQ formation was not detected with polymorphonuclear neutrophils from two unrelated MPO-deficient patients. Results suggest that the highly diffusible stable gas NO could initiate nitrosation at sites of neutrophil infiltration. PMID- 15531584 TI - The Sec14 homology domain regulates the cellular distribution and transforming activity of the Rho-specific guanine nucleotide exchange factor Dbs. AB - Dbs is a Rho-specific guanine nucleotide exchange factor that was identified in a screen for proteins whose overexpression cause deregulated growth in murine fibroblasts. Dbs contains multiple recognizable motifs including a centrally located Rho-specific guanine nucleotide exchange factor domain, a COOH-terminal Src homology 3 domain, two spectrin-like repeats, and a recently identified NH(2) terminal Sec14 homology domain. The transforming potential of Dbs is substantially activated by the removal of inhibitory sequences that lie outside of the core catalytic sequences, and in this current study we mapped this inhibition to the Sec14 domain. Surprisingly removal of the NH(2) terminus did not alter the catalytic activity of Dbs in vivo but rather altered its subcellular distribution. Whereas full-length Dbs was distributed primarily in a perinuclear structure that coincides with a marker for the Golgi apparatus, removal of the Sec14 domain was associated with translocation of Dbs to the cell periphery where it accumulated within membrane ruffles and lamellipodia. However, translocation of Dbs and the concomitant changes in the actin cytoskeleton were not sufficient to fully activate Dbs transformation. The Sec14 domain also forms intramolecular contacts with the pleckstrin homology domain, and these contacts must also be relieved to achieve full transforming activity. Collectively these observations suggest that the Sec14 domain regulates Dbs transformation through at least two distinct mechanisms, neither of which appears to directly influence the in vivo exchange activity of the protein. PMID- 15531585 TI - Interaction between transcription elongation factors and mRNA 3'-end formation at the Saccharomyces cerevisiae GAL10-GAL7 locus. AB - Spt6 is a conserved transcription factor that associates with RNA polymerase II (pol II) during elongation. Spt6 is essential for viability in Saccharomyces cerevisiae and regulates chromatin structure during pol II transcription. Here we present evidence that mutations that impair Spt6, a second elongation factor, Spt4, and pol II can affect 3'-end formation at GAL10. Additional analysis suggests that Spt6 is required for cotranscriptional association of the factor Ctr9, a member of the Paf1 complex, with GAL10 and GAL7, and that Ctr9 association with chromatin 3' of GAL10 is regulated by the GAL10 polyadenylation signal. Overall, these results provide new evidence for a connection between the transcription elongation factor Spt6 and 3'-end formation in vivo. PMID- 15531586 TI - Structure of the ubiquitin hydrolase UCH-L3 complexed with a suicide substrate. AB - Ubiquitin C-terminal hydrolases (UCHs) comprise a family of small ubiquitin specific proteases of uncertain function. Although no cellular substrates have been identified for UCHs, their highly tissue-specific expression patterns and the association of UCH-L1 mutations with human disease strongly suggest a critical role. The structure of the yeast UCH Yuh1-ubiquitin aldehyde complex identified an active site crossover loop predicted to limit the size of suitable substrates. We report the 1.45 A resolution crystal structure of human UCH-L3 in complex with the inhibitor ubiquitin vinylmethylester, an inhibitor that forms a covalent adduct with the active site cysteine of ubiquitin-specific proteases. This structure confirms the predicted mechanism of the inhibitor and allows the direct comparison of a UCH family enzyme in the free and ligand-bound state. We also show the efficient hydrolysis by human UCH-L3 of a 13-residue peptide in isopeptide linkage with ubiquitin, consistent with considerable flexibility in UCH substrate size. We propose a model for the catalytic cycle of UCH family members which accounts for the hydrolysis of larger ubiquitin conjugates. PMID- 15531587 TI - KLF11-mediated repression antagonizes Sp1/sterol-responsive element-binding protein-induced transcriptional activation of caveolin-1 in response to cholesterol signaling. AB - Cholesterol is a potent regulator of gene expression via a canonical pathway co regulated by SREBP and Sp1. Here we establish the caveolin-1 gene promoter as a cell type-specific model for SREBP/Sp1 regulation whereby lipoprotein cholesterol depletion activates caveolin-1 transcription in endothelial type cells, but not in fibroblasts, both in vitro and in vivo. By extending this model, we describe a novel pathway distinct from the prototypical SREBP/Sp1 regulatory loop involving the Sp1-like protein, KLF11. Through a combination of RNA interference, chromatin immunoprecipitation assays, electrophoretic mobility shift assays, and reporter assays, we demonstrate that in the presence of cholesterol, KLF11 acts as a dominant repressor of the caveolin-1 gene. Mechanistically, cholesterol depletion results in displacement of KLF11 from an Sp1 site flanking an SRE, indicating that activation by SREBP/Sp1 requires antagonism of KLF11 repression. The displacement of KLF11 results from both a down-regulation of its expression and competition by Sp1 for DNA binding. Therefore, these studies identify a novel pathway whereby KLF11 repression is coordinated with Sp1/SREBP activation of cholesterol-dependent gene expression in a cell type-specific manner and outline the mechanisms by which these functions are achieved. PMID- 15531588 TI - Antiviral activity of CYC202 in HIV-1-infected cells. AB - There are currently 40 million individuals in the world infected with human immunodeficiency virus (HIV). The introduction of highly active antiretroviral therapy (HAART) has led to a significant reduction in AIDS-related morbidity and mortality. Unfortunately, up to 25% of patients discontinue their initial HAART regimen. Current HIV-1 inhibitors target the fusion of the virus to the cell and two viral proteins, reverse transcriptase and protease. Here, we examined whether other targets, such as an activated transcription factor, could be targeted to block HIV-1 replication. We specifically asked whether we could target a cellular kinase needed for HIV-1 transcription using CYC202 (R-roscovitine), a pharmacological cyclin-dependent kinase inhibitor. We targeted the cdk2-cyclin E complex in HIV-1-infected cells because both cdk2 and cyclin E are nonessential during mammalian development and are likely replaced by other kinases. We found that CYC202 effectively inhibits wild type and resistant HIV-1 mutants in T cells, monocytes, and peripheral blood mononuclear cells at a low IC(50) and sensitizes these cells to enhanced apoptosis resulting in a dramatic drop in viral titers. Interestingly, the effect of CYC202 is independent of cell cycle stage and more specific for the cdk2-cyclin E complex. Finally, we show that cdk2 cyclin E is loaded onto the HIV-1 genome in vivo and that CYC202 is able to inhibit the uploading of this cdk-cyclin complex onto HIV-1 DNA. Therefore, targeting cellular enzymes necessary for HIV-1 transcription, which are not needed for cell survival, is a compelling strategy to inhibit wild type and mutant HIV-1 strains. PMID- 15531589 TI - Clostridium difficile toxin A induces expression of the stress-induced early gene product RhoB. AB - Clostridium difficile toxin A monoglucosylates the Rho family GTPases Rho, Rac, and Cdc42. Glucosylation leads to the functional inactivation of Rho GTPases and causes disruption of the actin cytoskeleton. A cDNA microarray revealed the immediate early gene rhoB as the gene that was predominantly up-regulated in colonic CaCo-2 cells after treatment with toxin A. This toxin A effect was also detectable in epithelial cells such as HT29 and Madin-Darby canine kidney cells, as well as NIH 3T3 fibroblasts. The expression of RhoB was time-dependent and correlated with the morphological changes of cells. The up-regulation of RhoB was approximately 15-fold and was based on the de novo synthesis of the GTPase because cycloheximide completely inhibited the toxin A effect. After 8 h, a steady state was reached, with no further increase in RhoB. The p38 MAPK inhibitor SB202190 reduced the expression of RhoB, indicating a participation of the p38 MAPK in this stress response. Surprisingly, newly formed RhoB protein was only partially glucosylated by toxin A, sparing a pool of potentially active RhoB, as checked by sequential C3bot-catalyzed ADP-ribosylation. A pull-down assay in fact revealed a significant amount of active RhoB in toxin A-treated cells that was not present in control cells. We demonstrate for the first time that toxin A has not only the property to inactivate the GTPases RhoA, Rac1, and Cdc42 by glucosylation, but it also has the property to generate active RhoB that likely contributes to the overall picture of toxin treatment. PMID- 15531590 TI - A structural model of the plant acyl-acyl carrier protein thioesterase FatB comprises two helix/4-stranded sheet domains, the N-terminal domain containing residues that affect specificity and the C-terminal domain containing catalytic residues. AB - Plant acyl-acyl carrier protein thioesterases (TEs) terminate the acyl-acyl carrier protein track of fatty acid biosynthesis and play an essential role in determining the amount and composition of fatty acids entering the storage lipid pool. A combination of bioinformatics tools was used to predict a three dimensional model for Arabidopsis FatB (AtFatB), which comprises a fold similar to that of Escherichia coli TEII, an enzyme that is functionally similar to plant TEs but lacks significant sequence similarity and displays different inhibitor sensitivity. The catalytic residues in AtFatB, Cys-264 and His-229, localize to the same region of the model as catalytic residues found in other enzymes with helix/multi-stranded sheet motifs (hot dog folds). Based on the model, we identified Asn-227 as a possible third member of the proposed papain-like catalytic triad. The conversion of Asn-227 to Ala resulted in a loss of detectable activity (>200-fold reduction), similar to the result seen for the equivalent mutation in papain. Mapping of plant TE specificity-affecting mutations onto the structural model showed that these mutations all cluster around the catalytic triad. Also, superposition of the crystallographically determined structures of the complexes of 4-hydroxybenzoyl-CoA TE with substrate and beta-hydroxydecanoyl thiol ester dehydrase with inhibitor onto the AtFatB model showed that the substrate and inhibitor localize to the same region as the AtFatB catalytic triad in their respective structures. Together these data corroborate the structural model and show that the hot dog fold is common to enzymes from both prokaryotes and eukaryotes and that this fold supports at least three different catalytic mechanisms. PMID- 15531591 TI - Evidence that the fosfomycin target Cys115 in UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) is essential for product release. AB - MurA (UDP-N-acetylglucosamine enolpyruvyl transferase, EC 2.5.1.7) is an essential enzyme in the biosynthesis of the peptidoglycan layer of the bacterial cell. It provides an attractive template for the design of novel antibiotic drugs and is the target of the naturally occurring antibiotic fosfomycin, which covalently attaches to Cys115 in the active site of the enzyme. Mutations of Cys115 to Asp exist in pathogens such as Mycobacteria or Chlamydia rendering these organisms resistant to fosfomycin. Thus, there is a need for the elucidation of the role of this cysteine in the MurA reaction. We determined the x-ray structure of the C115S mutant of Enterobacter cloacae MurA, which was crystallized in the presence of the substrates of MurA. The structure depicts the product state of the enzyme with enolpyruvyl-UDP-N-acetylglucosamine and inorganic phosphate trapped in the active site. Kinetic analysis revealed that the Cys-to-Ser mutation results in an enzyme that appears to perform a single turnover of the reaction. Opposing the common view of Cys115 as a key residue in the chemical reaction of enolpyruvyl transfer, we now conclude that the wild-type cysteine is essential for product release only. On the basis of a detailed comparison of the product state with the intermediate state and an unliganded state of MurA, we propose that dissociation of the products is an ordered event with inorganic phosphate leaving first. Phosphate departure appears to trigger a suite of conformational changes, which finally leads to opening of the two-domain structure of MurA and the release of the second product enolpyruvyl-UDP-N acetylglucosamine. PMID- 15531592 TI - Angiotensin I-converting enzyme ACE 2350*G and ACE-240*T-related genotypes and alleles are associated with higher susceptibility to endometriosis. AB - Endometriosis displays features similar to malignancy, ranging from neovascularization to local invasion and aggressive spread to distant organs. The altered vascular-related genes might be related to the development of endometriosis. This study investigates whether angiotensin I-converting enzyme (ACE) *A2350G and A-240T gene polymorphisms could be used as markers of susceptibility in endometriosis. Women were divided into two groups: (1) endometriosis group (n=150) and (2) non-endometriosis group (n=159). Genomic DNA was obtained from peripheral leukocytes. ACE A2350G and A-240T gene polymorphisms were amplified by PCR and detected after restriction enzyme digestion with BstUI and XbaI. Genotypes and allelic frequencies in both groups were compared. We observed that genotype distribution and allele frequency of ACE 2350 and ACE-240 gene polymorphisms in both groups were significantly different. Proportions of ACE 2350*A homozygote/heterozygote/G homozygote in both groups were: (1) 66.7/29.3/4% and (2) 96.2/3.1/0.7%. Proportions of ACE-240*A homozygote/heterozygote/T homozygote in both groups were: (1) 43.3/46/10.7% and (2) 62.9/35.8/1.3%. We concluded that ACE 2350*G and ACE-240*T-related genotypes and alleles are associated with higher susceptibility to endometriosis. ACE A2350G and A-240T gene polymorphisms might be associated with endometriosis development. PMID- 15531593 TI - Polymorphisms of the genes encoding the GSTM1, GSTT1 and GSTP1 in Korean women: no association with endometriosis. AB - Endometriosis, one of the most common gynaecologic disorders, shows significantly elevated prevalence in industrial areas and there is also a possible genetic predisposition. Glutathione-S-transferases (GSTs) are enzymes involved in the metabolism of many disease-causing carcinogens and mutagens that are present in human environments. An association between the incidence of endometriosis and the GST genotypes of patients has been suggested. The objective of the present study was to investigate whether the polymorphisms of GSTM1, GSTT1 and GSTP1 are related to endometriosis. Blood samples were available from 259 controls and 194 patients with advanced endometriosis diagnosed by both pathology and laparoscopic findings. The proportion of the GSTM1, GSTT1 and GSTP1 genotypes of the control group were comparable to other populations. There was no significant evidence that the distribution of the GSTM1 and GSTT1 genotype differed between the patients and the controls, with an allelic odds ratio (OR)=1.074 [95% confidence interval (CI)=0.737-1.564] and 1.239 (95% CI = 0.853-1.799), respectively. Also, there was no significant difference in the proportion of GSTP1 genotypes between the women with endometriosis and the control group with the OR = 0.823 (95% CI = 0.536-1.264). The higher risk alleles were contended as GSTM1, GSTT1 null mutation and GSTP1 Ile105Ile polymorphism. There was no significant increase in the risk of endometriosis as the number of higher risk alleles of the GST family increased. In conclusion, our findings suggest that the GSTM1, GSTT1 and GSTP1 genetic polymorphisms are not associated with the development of endometriosis in Korean women. PMID- 15531594 TI - Antibiotic cycling or rotation: a systematic review of the evidence of efficacy. AB - Of the interventions designed to reduce antibiotic resistance rates in hospitals, one that is currently attracting considerable interest, particularly in the intensive care unit setting, is antibiotic cycling or rotation. Cycling is the scheduled rotation of one class of antibiotics with one or more different classes exhibiting comparable spectra of activity; in order to fulfil the definition, the cycle must be repeated. Following a search of the literature we identified 11 articles in which the authors claimed to have evaluated the efficacy of this intervention. Only four were suitable for review, but, owing to multiple methodological flaws and a lack of standardization, the results of these studies do not permit reliable conclusions regarding the efficacy of cycling. Further studies are therefore required in order to resolve this question. However, before such studies can be undertaken, there are a great many issues relating to cycling which must be addressed. For the time being, we advise against the routine implementation of this measure as a means of reducing antibiotic resistance rates. PMID- 15531595 TI - Comparison of assays for detection of agents causing membrane damage in Staphylococcus aureus. AB - OBJECTIVES: To develop a novel beta-galactosidase leakage assay for Staphylococcus aureus and to evaluate this alongside other simple methods for detection of agents that cause membrane damage in staphylococci. METHODS: Using a PCR-based approach, a derivative of S. aureus RN4220 was constructed carrying the Escherichia coli lacZ gene under the control of the strong staphylococcal promoter, cap1A. Leakage of beta-galactosidase (BG) from this strain was examined after exposure for 10 min to various membrane-damaging agents at 4 x MIC, using a fluorescence assay and the substrate 4-methylumbelliferyl-beta-d-galactoside. Other assays for membrane damage involving protoplast lysis (PL), leakage of material absorbing at 260 nm (OD) and ATP release as well as the BacLight (BL) assay were carried out using established methods. RESULTS: All the assays, with the exception of the PL assay, detected membrane damage induced by cetyltrimethylammonium bromide, nisin, clofazimine and protegrin IB-367. However, the ability to detect membrane damage induced by these agents differed between the assay systems. The assays also varied considerably in their signal-to-noise ratio, with the ATP assay providing values for nisin approaching 100-fold that of the control. CONCLUSIONS: The PL assay is unsuitable for detection of membrane damaging agents in S. aureus. The other assays, including the BG assay, detect membrane damage. The OD assay should be sufficient for most purposes since it is effective, rapid and cheap to perform. Studies requiring maximum sensitivity and discrimination should employ the ATP assay. PMID- 15531596 TI - Pharmacodynamic activity of telithromycin against macrolide-susceptible and macrolide-resistant Streptococcus pneumoniae simulating clinically achievable free serum and epithelial lining fluid concentrations. AB - BACKGROUND: The association between macrolide resistance mechanisms and ketolide bacteriological eradication of Streptococcus pneumoniae remains poorly studied. The present study, using an in vitro model, assessed telithromycin pharmacodynamic activity against macrolide-susceptible and macrolide-resistant S. pneumoniae simulating clinically achievable free serum and epithelial lining fluid (ELF) concentrations. MATERIALS AND METHODS: Two macrolide-susceptible [PCR negative for both mef(A) and erm(B)] and six macrolide-resistant [five mef(A) positive/erm(B)-negative displaying various degrees of macrolide resistance and one mef(A)-negative/erm(B)-positive] S. pneumoniae were tested. Telithromycin was modelled simulating a dosage of 800 mg by mouth once daily [free serum: maximum concentration (C(max)) 0.7 mg/L, t(1/2) 10 h; and free ELF: C(max) 6.0 mg/L, t(1/2) 10 h]. Starting inocula were 1 x 10(6) cfu/mL in Mueller-Hinton broth with 2% lysed horse blood. Sampling at 0, 2, 4, 6, 12, 24 and 48 h assessed the extent of bacterial killing (decrease in log(10) cfu/mL versus initial inoculum). RESULTS: Telithromycin free serum concentrations achieved in the model were: C(max) 0.9+/-0.08 mg/L, AUC(0-24) 6.4+/-1.5 mg.h/L and t(1/2) of 10.6+/-1.6 h. Telithromycin free ELF concentrations achieved in the model were: C(max) 6.6+/ 0.8 mg/L, AUC(0-24) 45.5+/-5.5 mg.h/L and t(1/2) of 10.5+/-1.7 h. At 2 h, free serum telithromycin concentrations achieved a 1.0-1.9 log(10) reduction in inoculum compared with a 3.0-3.3 log(10) reduction with free ELF versus macrolide susceptible and macrolide-resistant S. pneumoniae. Free telithromycin serum and ELF concentrations simulating C(max)/MIC > or =14.1 and area under the curve to MIC (AUC(0-24)/MIC) > or =100 [time above the MIC (t > MIC) of 100%], were bactericidal (> or =3 log(10) killing) at 4, 6, 12, 24 and 48 h versus macrolide susceptible and macrolide-resistant S. pneumoniae. CONCLUSION: Telithromycin serum and ELF concentrations rapidly eradicated macrolide-susceptible and macrolide-resistant S. pneumoniae regardless of resistance phenotype. Achieving C(max)/MIC > or =14.1 and AUC(0-24)/MIC > or =100 resulted in bactericidal activity at 4 h with no regrowth over 48 h. PMID- 15531597 TI - Emergence of invasive erythromycin-resistant Streptococcus pneumoniae strains in Portugal: contribution and phylogenetic relatedness of serotype 14. AB - OBJECTIVES: To study the phenotype and phylogenetic relatedness of invasive Streptococcus pneumoniae strains isolated in Portugal. METHODS: A total of 614 invasive S. pneumoniae strains, isolated in Portugal from 1999 to the first 6 months of 2002, were characterized using serotyping and antimicrobial susceptibility testing. National outpatient sale data for macrolides were compared with erythromycin resistance. We investigated the main clonal lineages from erythromycin-resistant strains of serotype 14 by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). RESULTS: The emergence of erythromycin-resistant strains correlated well with the usage of azithromycin in Portugal during the period of the study (r=0.900, P=0.001). Serotype 14 made the largest contribution to this emergence. We found two clonal complexes (CC) among erythromycin-resistant strains: CC1 was formed by three sequence types (ST), ST156, ST143 and ST1042, and CC2 by ST15 and ST9. All STs described, except ST1042, are putative founders of clonal groups or subgroups from serotype 14 as defined in the pneumococcal MLST database. CONCLUSIONS: This study suggests that macrolides usage is an important factor enhancing the spread of invasive erythromycin-resistant S. pneumoniae clones in Portugal. The study also makes a contribution to the understanding of spread of erythromycin resistant clones in an international context. PMID- 15531598 TI - Trends of penicillin and erythromycin resistance among invasive Streptococcus pneumoniae in Europe. AB - OBJECTIVES: To forecast trends in resistance to penicillin and erythromycin among Streptococcus pneumoniae in Europe. METHODS: Since 1999, the European Antimicrobial Resistance Surveillance System (EARSS) has collected routine antimicrobial susceptibility test results of S. pneumoniae. To observe and predict changes of reduced susceptibility over time, we used a multinomial logistic regression model. RESULTS: Large variations in penicillin and erythromycin non-susceptibility were observed between countries, and reduced susceptibility to erythromycin (17%) has become more frequent than reduced susceptibility to penicillin (10%) in Europe overall. An overall decrease in single penicillin non-susceptibility, but an increase in dual non-susceptibility was observed, indicating a shift of single penicillin to combined non susceptibility with erythromycin. By 2006, the proportion of single erythromycin and dual non-susceptibility could increase to as much as 20.4% and 8.9%, respectively. CONCLUSIONS: Our results indicate that appropriately dosed beta lactams for empirical therapy are still the treatment of choice, and that macrolides should be used with prudence. PMID- 15531599 TI - Accurate identification of alternatively spliced exons using support vector machine. AB - MOTIVATION: Alternative splicing is a major component of the regulatory action on mammalian transcriptomes. It is estimated that over half of all human genes have more than one splice variant. Previous studies have shown that alternatively spliced exons possess several features that distinguish them from constitutively spliced ones. Recently, we have demonstrated that such features can be used to distinguish alternative from constitutive exons. In the current study, we used advanced machine learning methods to generate robust classifier of alternative exons. RESULTS: We extracted several hundred local sequence features of constitutive as well as alternative exons. Using feature selection methods we find seven attributes that are dominant for the task of classification. Several less informative features help to slightly increase the performance of the classifier. The classifier achieves a true positive rate of 50% for a false positive rate of 0.5%. This result enables one to reliably identify alternatively spliced exons in exon databases that are believed to be dominated by constitutive exons. PMID- 15531600 TI - Intervention in context-sensitive probabilistic Boolean networks. AB - MOTIVATION: Intervention in a gene regulatory network is used to help it avoid undesirable states, such as those associated with a disease. Several types of intervention have been studied in the framework of a probabilistic Boolean network (PBN), which is essentially a finite collection of Boolean networks in which at any discrete time point the gene state vector transitions according to the rules of one of the constituent networks. For an instantaneously random PBN, the governing Boolean network is randomly chosen at each time point. For a context-sensitive PBN, the governing Boolean network remains fixed for an interval of time until a binary random variable determines a switch. The theory of automatic control has been previously applied to find optimal strategies for manipulating external (control) variables that affect the transition probabilities of an instantaneously random PBN to desirably affect its dynamic evolution over a finite time horizon. This paper extends the methods of external control to context-sensitive PBNs. RESULTS: This paper treats intervention via external control variables in context-sensitive PBNs by extending the results for instantaneously random PBNs in several directions. First, and most importantly, whereas an instantaneously random PBN yields a Markov chain whose state space is composed of gene vectors, each state of the Markov chain corresponding to a context-sensitive PBN is composed of a pair, the current gene vector occupied by the network and the current constituent Boolean network. Second, the analysis is applied to PBNs with perturbation, meaning that random gene perturbation is permitted at each instant with some probability. Third, the (mathematical) influence of genes within the network is used to choose the particular gene with which to intervene. Lastly, PBNs are designed from data using a recently proposed inference procedure that takes steady-state considerations into account. The results are applied to a context-sensitive PBN derived from gene-expression data collected in a study of metastatic melanoma, the intent being to devise a control strategy that reduces the WNT5A gene's action in affecting biological regulation, since the available data suggest that disruption of this influence could reduce the chance of a melanoma metastasizing. PMID- 15531601 TI - Non-sequential structure-based alignments reveal topology-independent core packing arrangements in proteins. AB - MOTIVATION: Proteins of the same class often share a secondary structure packing arrangement but differ in how the secondary structure units are ordered in the sequence. We find that proteins that share a common core also share local sequence-structure similarities, and these can be exploited to align structures with different topologies. In this study, segments from a library of local sequence-structure alignments were assembled hierarchically, enforcing the compactness and conserved inter-residue contacts but not sequential ordering. Previous structure-based alignment methods often ignore sequence similarity, local structural equivalence and compactness. RESULTS: The new program, SCALI (Structural Core ALIgnment), can efficiently find conserved packing arrangements, even if they are non-sequentially ordered in space. SCALI alignments conserve remote sequence similarity and contain fewer alignment errors. Clustering of our pairwise non-sequential alignments shows that recurrent packing arrangements exist in topologically different structures. For example, the three-layer sandwich domain architecture may be divided into four structural subclasses based on internal packing arrangements. These subclasses represent an intermediate level of structure classification, more general than topology, but more specific than architecture as defined in CATH. A strategy is presented for developing a set of predictive hidden Markov models based on multiple SCALI alignments. PMID- 15531602 TI - A simple statistical method for discriminating outer membrane proteins with better accuracy. AB - MOTIVATION: Discriminating outer membrane proteins from other folding types of globular and membrane proteins is an important task both for identifying outer membrane proteins from genomic sequences and for the successful prediction of their secondary and tertiary structures. RESULTS: We have systematically analyzed the amino acid composition of globular proteins from different structural classes and outer membrane proteins. We found that the residues, Glu, His, Ile, Cys, Gln, Asn and Ser, show a significant difference between globular and outer membrane proteins. Based on this information, we have devised a statistical method for discriminating outer membrane proteins from other globular and membrane proteins. Our approach correctly picked up the outer membrane proteins with an accuracy of 89% for the training set of 337 proteins. On the other hand, our method has correctly excluded the globular proteins at an accuracy of 79% in a non-redundant dataset of 674 proteins. Furthermore, the present method is able to correctly exclude alpha-helical membrane proteins up to an accuracy of 80%. These accuracy levels are comparable to other methods in the literature, and this is a simple method, which could be used for dissecting outer membrane proteins from genomic sequences. The influence of protein size, structural class and specific residues for discrimination is discussed. PMID- 15531603 TI - Protein homology detection by HMM-HMM comparison. AB - MOTIVATION: Protein homology detection and sequence alignment are at the basis of protein structure prediction, function prediction and evolution. RESULTS: We have generalized the alignment of protein sequences with a profile hidden Markov model (HMM) to the case of pairwise alignment of profile HMMs. We present a method for detecting distant homologous relationships between proteins based on this approach. The method (HHsearch) is benchmarked together with BLAST, PSI-BLAST, HMMER and the profile-profile comparison tools PROF_SIM and COMPASS, in an all against-all comparison of a database of 3691 protein domains from SCOP 1.63 with pairwise sequence identities below 20%.Sensitivity: When the predicted secondary structure is included in the HMMs, HHsearch is able to detect between 2.7 and 4.2 times more homologs than PSI-BLAST or HMMER and between 1.44 and 1.9 times more than COMPASS or PROF_SIM for a rate of false positives of 10%. Approximately half of the improvement over the profile-profile comparison methods is attributable to the use of profile HMMs in place of simple profiles. Alignment quality: Higher sensitivity is mirrored by an increased alignment quality. HHsearch produced 1.2, 1.7 and 3.3 times more good alignments ('balanced' score >0.3) than the next best method (COMPASS), and 1.6, 2.9 and 9.4 times more than PSI-BLAST, at the family, superfamily and fold level, respectively.Speed: HHsearch scans a query of 200 residues against 3691 domains in 33 s on an AMD64 2GHz PC. This is 10 times faster than PROF_SIM and 17 times faster than COMPASS. PMID- 15531604 TI - A Java tool for dynamic web-based 3D visualization of anatomy and overlapping gene or protein expression patterns. AB - The Gene Expression Viewer is a web-launched three-dimensional visualization tool, tailored to compare surface reconstructions of multi-channel image volumes generated by confocal microscopy or micro-CT. PMID- 15531605 TI - OMGProm: a database of orthologous mammalian gene promoters. AB - SUMMARY: Sequence comparisons between human and rodents are increasingly being used for the identification of gene regulatory regions. The effectiveness of such an approach largely depends on the quality and availability of promoter sequences. We developed OMGProm by integrating three data sources: (1) experimentally supported full-length cDNA, promoter and first exon sequences; (2) homology information from HomoloGene and (3) the human and mouse genomic sequences. The current version of OMGProm contains 8550 promoter pairs of 6373 orthologous human and mouse genes, where supporting experimental evidence for transcription start site annotation exists in at least one species. PMID- 15531606 TI - Sensitivity, principal component and flux analysis applied to signal transduction: the case of epidermal growth factor mediated signaling. AB - MOTIVATION: Novel high-throughput genomic and proteomic tools are allowing the integration of information from a range of biological assays into a single conceptual framework. This framework is often described as a network of biochemical reactions. We present strategies for the analysis of such networks. RESULTS: The direct differential method is described for the systematic evaluation of scaled sensitivity coefficients in reaction networks. Principal component analysis, based on an eigenvalue-eigenvector analysis of the scaled sensitivity coefficient matrix, is applied to rank individual reactions in the network based on their effect on system output. When combined with flux analysis, sensitivity analysis allows model reduction or simplification. Using epidermal growth factor (EGF) mediated signaling and trafficking as an example of signal transduction, we demonstrate that sensitivity analysis quantitatively reveals the dependence of dual-phosphorylated extracellular signal-regulated kinase (ERK) concentration on individual reaction rate constants. It predicts that EGF mediated reactions proceed primarily via an Shc-dependent pathway. Further, it suggests that receptor internalization and endosomal signaling are important features regulating signal output only at low EGF dosages and at later times. PMID- 15531608 TI - The MIPS mammalian protein-protein interaction database. AB - SUMMARY: The MIPS mammalian protein-protein interaction database (MPPI) is a new resource of high-quality experimental protein interaction data in mammals. The content is based on published experimental evidence that has been processed by human expert curators. We provide the full dataset for download and a flexible and powerful web interface for users with various requirements. PMID- 15531607 TI - Extracting multiple structural alignments from pairwise alignments: a comparison of a rigorous and a heuristic approach. AB - MOTIVATION: Multiple structural alignments (MSTAs) provide position-specific information on the sequence variability allowed by protein folds. This information can be exploited to better understand the evolution of proteins and the physical chemistry of polypeptide folding. Most MSTA methods rely on a pre computed library of pairwise alignments. This library will in general contain conflicting residue equivalences not all of which can be realized in the final MSTA. Hence to build a consistent MSTA, these methods have to select a conflict free subset of equivalences. RESULTS: Using a dataset with 327 families from SCOP 1.63 we compare the ability of two different methods to select an optimal conflict-free subset of equivalences. One is an implementation of Reinert et al.'s integer linear programming formulation (ILP) of the maximum weight trace problem (Reinert et al., 1997, Proc. 1st Ann. Int. Conf. Comput. Mol. Biol. (RECOMB-97), ACM Press, New York). This ILP formulation is a rigorous approach but its complexity is difficult to predict. The other method is T-Coffee (Notredame et al., 2000) which uses a heuristic enhancement of the equivalence weights which allow it to use the speed and simplicity of the progressive alignment approach while still incorporating information of all alignments in each step of building the MSTA. We find that although the ILP formulation consistently selects a more optimal set of conflict-free equivalences, the differences are small and the quality of the resulting MSTAs are essentially the same for both methods. Given its speed and predictable complexity, our results show that T-Coffee is an attractive alternative for producing high-quality MSTAs. PMID- 15531609 TI - Classification using partial least squares with penalized logistic regression. AB - MOTIVATION: One important aspect of data-mining of microarray data is to discover the molecular variation among cancers. In microarray studies, the number n of samples is relatively small compared to the number p of genes per sample (usually in thousands). It is known that standard statistical methods in classification are efficient (i.e. in the present case, yield successful classifiers) particularly when n is (far) larger than p. This naturally calls for the use of a dimension reduction procedure together with the classification one. RESULTS: In this paper, the question of classification in such a high-dimensional setting is addressed. We view the classification problem as a regression one with few observations and many predictor variables. We propose a new method combining partial least squares (PLS) and Ridge penalized logistic regression. We review the existing methods based on PLS and/or penalized likelihood techniques, outline their interest in some cases and theoretically explain their sometimes poor behavior. Our procedure is compared with these other classifiers. The predictive performance of the resulting classification rule is illustrated on three data sets: Leukemia, Colon and Prostate. PMID- 15531610 TI - CGH-Explorer: a program for analysis of array-CGH data. AB - SUMMARY: CGH-Explorer is a program for visualization and statistical analysis of microarray-based comparative genomic hybridization (array-CGH) data. The program has preprocessing facilities, tools for graphical exploration of individual arrays or groups of arrays, and tools for statistical identification of regions of amplification and deletion. PMID- 15531611 TI - GoArrays: highly dynamic and efficient microarray probe design. AB - MOTIVATION: The use of oligonucleotide microarray technology requires a very detailed attention to the design of specific probes spotted on the solid phase. These problems are far from being commonplace since they refer to complex physicochemical constraints. Whereas there are more and more publicly available programs for microarray oligonucleotide design, most of them use the same algorithm or criteria to design oligos, with only little variation. RESULTS: We show that classical approaches used in oligo design software may be inefficient under certain experimental conditions, especially when dealing with complex target mixtures. Indeed, our biological model is a human obligate parasite, the microsporidia Encephalitozoon cuniculi. Targets that are extracted from biological samples are composed of a mixture of pathogen transcripts and host cell transcripts. We propose a new approach to design oligonucleotides which combines good specificity with a potentially high sensitivity. This approach is original in the biological point of view as well as in the algorithmic point of view. We also present an experimental validation of this new strategy by comparing results obtained with standard oligos and with our composite oligos. A specific E.cuniculi microarray will overcome the difficulty to discriminate the parasite mRNAs from the host cell mRNAs demonstrating the power of the microarray approach to elucidate the lifestyle of an intracellular pathogen using mix mRNAs. PMID- 15531612 TI - In silico gene function prediction using ontology-based pattern identification. AB - MOTIVATION: With the emergence of genome-wide expression profiling data sets, the guilt by association (GBA) principle has been a cornerstone for deriving gene functional interpretations in silico. Given the limited success of traditional methods for producing clusters of genes with great amounts of functional similarity, new data-mining algorithms are required to fully exploit the potential of high-throughput genomic approaches. RESULTS: Ontology-based pattern identification (OPI) is a novel data-mining algorithm that systematically identifies expression patterns that best represent existing knowledge of gene function. Instead of relying on a universal threshold of expression similarity to define functionally related groups of genes, OPI finds the optimal analysis settings that yield gene expression patterns and gene lists that best predict gene function using the principle of GBA. We applied OPI to a publicly available gene expression data set on the life cycle of the malarial parasite Plasmodium falciparum and systematically annotated genes for 320 functional categories based on current Gene Ontology annotations. An ontology-based hierarchical tree of the 320 categories provided a systems-wide biological view of this important malarial parasite. PMID- 15531613 TI - The SBW-MATLAB interface. AB - SUMMARY: The SBW-MATLAB Interface allows MATLAB users to take advantage of the wide variety of tools available through SBW, the Systems Biology Workbench (Sauro et al. (2003) OMICS, 7, 355-372). It also enables MATLAB users to themselves create SBW-enabled tools which can be freely distributed. PMID- 15531614 TI - An alternative model of amino acid replacement. AB - MOTIVATION: The observed correlations between pairs of homologous protein sequences are typically explained in terms of a Markovian dynamic of amino acid substitution. This model assumes that every location on the protein sequence has the same background distribution of amino acids, an assumption that is incompatible with the observed heterogeneity of protein amino acid profiles and with the success of profile multiple sequence alignment. RESULTS: We propose an alternative model of amino acid replacement during protein evolution based upon the assumption that the variation of the amino acid background distribution from one residue to the next is sufficient to explain the observed sequence correlations of homologs. The resulting dynamical model of independent replacements drawn from heterogeneous backgrounds is simple and consistent, and provides a unified homology match score for sequence-sequence, sequence-profile and profile-profile alignment. PMID- 15531615 TI - Predicting gene function through systematic analysis and quality assessment of high-throughput data. AB - MOTIVATION: Determining gene function is an important challenge arising from the availability of whole genome sequences. Until recently, approaches based on sequence homology were the only high-throughput method for predicting gene function. Use of high-throughput generated experimental data sets for determining gene function has been limited for several reasons. RESULTS: Here a new approach is presented for integration of high-throughput data sets, leading to prediction of function based on relationships supported by multiple types and sources of data. This is achieved with a database containing 125 different high-throughput data sets describing phenotypes, cellular localizations, protein interactions and mRNA expression levels from Saccharomyces cerevisiae, using a bit-vector representation and information content-based ranking. The approach takes characteristic and qualitative differences between the data sets into account, is highly flexible, efficient and scalable. Database queries result in predictions for 543 uncharacterized genes, based on multiple functional relationships each supported by at least three types of experimental data. Some of these are experimentally verified, further demonstrating their reliability. The results also generate insights into the relative merits of different data types and provide a coherent framework for functional genomic datamining. AVAILABILITY: Free availability over the Internet. CONTACT: f.c.p.holstege@med.uu.nl SUPPLEMENTARY INFORMATION: http://www.genomics.med.uu.nl/pub/pk/comb_gen_network. PMID- 15531616 TI - ESTminer: a suite of programs for gene and allele identification. AB - ESTminer is a collection of programs that use expressed sequence tag (EST) data from inbred genomes to identify unique genes within gene families. The algorithm utilizes Cap3 to perform an initial clustering of related EST sequences to produce a consensus sequence of a gene family. These consensus sequences are then used to collect all ESTs in the original EST library that are related using BLAST. A redundancy based criterion is applied to each EST to identify reliable unique gene-sequences. Using a highly inbred genome as a source of ESTs eliminates the necessity of computing covariance on each polymorphism to identify alleles of the same gene, thus making this algorithm more streamlined than other alternatives which must computationally attempt to distinguish genes from alleles. AVAILABILITY: The programs were written in PERL and are freely available at http://www.soybase.org/publication_data/Nelson/ESTminer/ESTminer.html CONTACT: nelsonrt@iastate.edu SUPPLEMENTARY INFORMATION: Figures and dataset can be obtained from: http://www.soybase.org/publication_data/Nelson/ESTminer/ESTminer.html. PMID- 15531617 TI - ISYMOD: a knowledge warehouse for the identification, assembly and analysis of bacterial integrated systems. AB - MOTIVATION: Complex biological functions emerge from interactions between proteins in stable supra-molecular assemblies and/or through transitory contacts. Most of the time protein partners of the assemblies are composed of one or several domains which exhibit different biochemical functions. Thus the study of cellular process requires the identification of different functional units and their integration in an interaction network; such complexes are referred to as integrated systems. In order to exploit with optimum efficiency the increased release of data, automated bioinformatics strategies are needed to identify, reconstruct and model such systems. For that purpose, we have developed a knowledge warehouse dedicated to the representation and acquisition of bacterial integrated systems involved in the exchange of the bacterial cell with its environment. RESULTS: ISYMOD is a knowledge warehouse that consistently integrates in the same environment the data and the methods used for their acquisition. This is achieved through the construction of (1) a domain knowledge base (DKB) devoted to the storage of the knowledge about the systems, their functional specificities, their partners and how they are related and (2) a methodological knowledge base (MKB) which depicts the task layout used to identify and reconstruct functional integrated systems. Instantiation of the DKB is obtained by solving the tasks of the MKB, whereas some tasks need instances of the DKB to be solved. AROM, an object-based knowledge representation system, has been used to design the DKB, and its task manager, AROMTasks, for developing the MKB. In this study two integrated systems, ABC transporters and two component systems, both involved in adaptation processes of a bacterial cell to its biotope, have been used to evaluate the feasibility of the approach. PMID- 15531618 TI - AUG codons at the beginning of protein coding sequences are frequent in eukaryotic mRNAs with a suboptimal start codon context. AB - MOTIVATION: The translation start site plays an important role in the control of translation efficiency of eukaryotic mRNAs. However, mRNAs with a suboptimal context of start AUG codon are relatively abundant. It is likely that at least some mRNAs with suboptimal start codon context contain the other signals providing additional information for efficient AUG recognition. RESULTS: Frequency of AUG codons at the beginning of the coding part of eukaryotic mRNAs was analyzed in relation to the context of translation start codon. It was found that the observed downstream AUG content in the mRNAs with optimal start codon context was close to the expected value, whereas it was significantly higher in the mRNAs with a suboptimal context. It is likely that downstream AUG codons can often be utilized as additional start sites to increase translation rate of mRNAs with a suboptimal context of the annotated start codon and many eukaryotic proteins can be characterized by some N-end heterogeneity. PMID- 15531619 TI - Randomized controlled trial of effects of the airflow through the upper respiratory tract of intubated brain-injured patients on brain temperature and selective brain cooling. AB - BACKGROUND: Pyrexia is common after brain injury; it is generally believed to affect outcome adversely and the usual clinical methods of reducing temperature are not effective. The normal physiological mechanisms of brain cooling are heat loss from the upper airways and through the skull, and these can produce selective brain cooling. METHODS: Air at room temperature and humidity was continuously administered to 15 brain-injured, intubated and mechanically ventilated patients via a sponge-tipped oxygen catheter in each nostril at a combined rate of 115 ml kg(-1) min(-1). Brain temperature was measured using a pressure-temperature Camino catheter which is designed to site the thermistor 1 cm into the parenchyma in the frontal lobe. Oesophageal temperature was measured using an oesophageal stethoscope with a thermistor. After establishing baseline for 30 min, patients were randomized to receive airflow or no airflow for 6 h and then crossed over for a further 6 h. RESULTS: Airflow replicating normal resting minute volume did not produce clinically relevant or statistically significant reductions in brain temperature [0.13 (SD 0.55) degrees C; 95% CI, 0.43-0.17 degrees C]. However, we serendipitously found some evidence of selective brain cooling via the skull, but this needs further substantiation. CONCLUSIONS: A flow of humidified air at room temperature through the upper respiratory tracts of intubated brain-injured patients did not produce clinically relevant or statistically significant reductions in brain temperature measured in the frontal lobe. PMID- 15531620 TI - Randomized controlled trial of the haemodynamic and recovery effects of xenon or propofol anaesthesia. AB - BACKGROUND: There is limited clinical experience with xenon in a large number of patients. We present intra- and postoperative haemodynamic and recovery data comparing xenon and total intravenous anaesthesia with propofol. METHODS: A total of 160 patients aged 18-60 years (ASA I and II) undergoing elective surgery took part in this prospective non-blinded randomized controlled trial. After local ethics committee approval and written informed consent, patients were allocated randomly to either the xenon or the propofol group. Anaesthesia was induced with propofol and remifentanil and was maintained with xenon at 60% (minimal alveolar concentration 0.95) or with propofol 0.1-0.12 mg kg(-1) min(-1). Remifentanil was titrated to clinical need in both groups. RESULTS: The two study groups were comparable with respect to age, weight, height, gender and ASA classification. Baseline in heart rate and systolic arterial pressure (SAP) were comparable in both groups. Following induction, SAP initially decreased but returned to baseline values over 15 min in the xenon group and differed significantly from the propofol group. Heart rate decreased significantly only in the xenon group and remained at stable values. Occurrence and duration of hypertension, hypotension and bradycardia showed no significant difference between groups. Patient recovery time in the post-anaesthetic care unit and recovery from anaesthesia was similar in the two groups. CONCLUSIONS: After induction the xenon/opioid regimen maintains systolic blood pressure at baseline levels and a low heart rate. No differences between groups were found in haemodynamic stability during anaesthesia. Recovery from xenon anaesthesia was similar to that observed in the propofol group. PMID- 15531621 TI - Chronic postoperative pain: the case of inguinal herniorrhaphy. PMID- 15531622 TI - Kidney dysfunction in the postoperative period. AB - The development of perioperative acute renal failure is associated with a high incidence of morbidity and mortality. Although this incidence varies with different surgical procedures and with the definition used for renal failure, we now understand better the aetiology of the underlying problem. However, successful strategies to provide renal protection or strategies for 'rescue therapy' are either lacking, unsubstantiated by randomized clinical trials, or show no significant efficacy. The present review considers the physiology and pharmacology of the kidney; the characterization of tests of renal function; the cause of postoperative renal dysfunction; what is presently available for its prevention and treatment; and the effect of postoperative renal impairment on patient outcome. PMID- 15531623 TI - Cerebral haemodynamics in patients with chronic renal failure: effects of haemodialysis. AB - BACKGROUND: We measured middle cerebral artery (MCA) flow velocity (FV), dynamic pressure autoregulation, and carbon dioxide reactivity (CRCO(2)) in patients with chronic renal failure before and after haemodialysis using transcranial Doppler ultrasonography. METHODS: Twelve patients on long-term haemodialysis were recruited. MCA FV was measured continuously. The transient hyperaemic response test was used to assess cerebral autoregulation, and per cent change in FV per kPa change in end-tidal carbon dioxide was calculated to assess CRCO(2). All measurements were recorded before and after haemodialysis. RESULTS: MCA FV (mean [sd]) decreased from 57 (10) cm s(-1) before to 46 (13) cm s(-1) after haemodialysis (P<0.01). The transient hyperaemic response ratio (THRR) was (mean [sd]) 1.29 (0.13) before haemodialysis and did not change significantly following haemodialysis (1.36 [0.10]). CRCO(2) was 21.7 (8.3)% kPa(-1) before haemodialysis and remained unchanged afterwards (20.9 [3.8]% kPa(-1)). Values in normal subjects for MCA FV, THRR and CRCO(2) are 56 (12) cm s(-1), 1.26 (0.13) and 22 (6)% kPa(-1), respectively. CONCLUSIONS: MCA FV decreases significantly after haemodialysis. Dynamic pressure autoregulation and CRCO(2) remain normal in patients with chronic renal failure, and are not altered significantly by haemodialysis. PMID- 15531624 TI - Short report: encouraging GPs to complete postal questionnaires--one big prize or many small prizes? A randomized controlled trial. AB - BACKGROUND: Low response rates to surveys are a problem in general practice. There is evidence that offering GPs incentives improves response rates to postal questionnaires. However, there is less evidence about the most effective form of incentive. OBJECTIVE: Our trial aimed to maximize response to a postal questionnaire and to test the most effective form of incentive. METHODS: The study involved a randomized controlled trial of a postal survey RESULTS: The incentive of a lottery for six bottles of champagne generated a response rate of 79%. Furthermore, one chance of six bottles generated 9% more responses than six chances of one bottle. CONCLUSIONS: This study has established that, among incentives for postal questionnaires, one big prize improves the yield more than many small prizes despite the lower odds of winning. It has also confirmed that offering a modest incentive to GPs generates good response rates for postal questionnaires. PMID- 15531625 TI - Complementary and alternative medicine use in patients with chronic diseases in primary care is associated with perceived quality of care and cultural beliefs. AB - OBJECTIVES: The purpose of our study was to determine the prevalence of complementary and alternative medicine (CAM) use and its clinical and psycho social correlates, including perceived satisfaction with care and cultural health beliefs. METHODS: A cross-sectional study was carried out in public sector primary care clinics in Singapore using a random sample of 488 adult patients with chronic diseases. The measures were CAM use, satisfaction with care and traditional health beliefs. RESULTS: The 1 year prevalence of CAM use was 22.7%. In univariate analyses, factors associated with CAM use included: middle age, arthritis, musculoskeletal disorders and stroke, multiple conditions, poor perceived health, family use of CAM, recommendation by close social contacts, strong adherence to traditional health beliefs and perceived satisfaction with care. Patients who were dissatisfied/very dissatisfied with the cost of treatment [odds ratio (OR) = 1.79, 95% confidence interval (CI) 1.15-2.82] and waiting time (OR = 1.96, 95% CI 1.20-3.19) were more likely to use CAM. Patients who were very satisfied with the benefit from treatment were much less likely to use CAM (OR = 0.49, 95% CI 0.29-0.83). Satisfaction with doctor-patient interaction was not associated with CAM use. Being 'very satisfied' on overall care satisfaction was significantly associated with much less CAM use (OR = 0.30, 95% CI 0.14-0.68). Multivariate analyses confirmed that CAM use was significantly and independently predicted by the 'chronic disease triad' (arthritis/musculoskeletal disorders/stroke) (OR = 4.08, 95% CI 2.45-6.83), overall satisfaction with care (OR = 0.32, 95% CI 0.14-0.74) and strong adherence to traditional health beliefs (OR = 1.88, 95% CI 1.07-3.31). CONCLUSION: CAM use in Asian patients is prevalent and associated with the 'chronic disease triad' (of arthritis, musculoskeletal disorders and stroke), satisfaction with care and cultural beliefs. In particular, CAM use is not associated with the quality of doctor-patient interaction. PMID- 15531626 TI - Cost-utility analysis of osteopathy in primary care: results from a pragmatic randomized controlled trial. AB - BACKGROUND: Spinal pain is common and costly to health services and society. Management guidelines have encouraged primary care referral for spinal manipulation, but the evidence base is weak. More economic evaluations alongside pragmatic trials have been recommended. OBJECTIVE: Our aim was to assess the cost utility of a practice-based osteopathy clinic for subacute spinal pain. METHODS: A cost-utility analysis was performed alongside a pragmatic single-centre randomized controlled trial in a primary care osteopathy clinic accepting referrals from 14 neighbouring practices in North West Wales. Patients with back pain of 2-12 weeks duration were randomly allocated to treatment with osteopathy plus usual GP care or usual GP care alone. Costs were measured from a National Health Service (NHS) perspective. All primary and secondary health care interventions recorded in GP notes were collected for the study period. We calculated quality adjusted life year (QALY) gains based on EQ-5D responses from patients in the trial, and then cost per QALY ratios. Confidence intervals (CIs) were estimated using non-parametric bootstrapping. RESULTS: Osteopathy plus usual GP care was more effective but resulted in more health care costs than usual GP care alone. The point estimate of the incremental cost per QALY ratio was 3560 pounds (80% CI 542 pounds-77,100 pounds). Sensitivity analysis examining spine related costs alone and total costs excluding outliers resulted in lower cost per QALY ratios. CONCLUSION: A primary care osteopathy clinic may be a cost-effective addition to usual GP care, but this conclusion was subject to considerable random error. Rigorous multi-centre studies are needed to assess the generalizability of this approach. PMID- 15531627 TI - A thermostable variant of fructose bisphosphate aldolase constructed by directed evolution also shows increased stability in organic solvents. AB - Thermostable variants of the Class II fructose bisphosphate aldolase have been isolated following four rounds of directed evolution using DNA shuffling of the fda genes from Escherichia coli and Edwardsiella ictaluri. Variants from all four generations of evolution have been purified and characterized. The variants show increased thermostability with no loss of catalytic function at room temperature. The temperature at which 50% of the initial enzyme activity is lost after incubation for 10 min (T50) of the most stable variant, 4-43D6, is increased by 11-12 degrees C over the wild-type enzymes and the half-life of activity at 53 degrees C is increased approximately 190-fold. In addition, variant 4-43D6 shows increased stability to treatment with organic solvents. DNA sequencing of the evolved variants has identified the mutations which have been introduced and which lead to increased thermostability, and the role of the mutations introduced is discussed. PMID- 15531628 TI - Rapid isolation of high-affinity protein binding peptides using bacterial display. AB - A robust bacterial display methodology was developed that allows the rapid isolation of peptides that bind to arbitrarily selected targets with high affinity. To demonstrate the utility of this approach, a large library (5 x 10(10) clones) was constructed composed of random 15-mer peptide insertions constrained within a flexible, surface exposed loop of the Escherichia coli outer membrane protein A (OmpA). The library was screened for binding to five unrelated proteins, including targets previously used in phage display selections: human serum albumin, anti-T7 epitope mAb, human C-reactive protein, HIV-1 GP120 and streptavidin. Two to four rounds of enrichment (2-4 days) were sufficient to enrich peptide ligands having high affinity for each of the target proteins. Strong amino acid consensus sequences were apparent for each of the targets tested, with up to seven consensus residues. Isolated peptide ligands remained functional when expressed as insertional fusions within a monomeric fluorescent protein. This bacterial display methodology provides an efficient process for identifying peptide affinity reagents and should be useful in a variety of molecular recognition applications. PMID- 15531629 TI - In silico evaluation of two mass spectrometry-based approaches for the identification of novel human leukocyte cell-surface proteins. AB - The identification and quantitation of cell-surface proteins expressed by leukocytes currently use the wide availability of monoclonal antibodies (mAb) in immunohistochemical and flow cytometric assays. Presently, approximately 400 such proteins have been characterized; however, analysis of the completed human genome sequence indicates that it may contain several thousand as-yet unidentified molecules, which may be expressed on the leukocyte cell surface. Recent advances in protein isolation and analysis using mass spectrometry illustrate that it is now feasible to identify the protein composition of a complex sample such as a plasma membrane extract. Such an approach may be useful for the identification of the cell-surface proteins that have not been identified using mAb techniques. Here, we detail the results of an in silico evaluation of the peptides isolated using two methods used to label plasma membrane proteins to determine whether these methods are suitable for the identification of known leukocyte cell-surface proteins by mass spectrometry. The labeling of cell-surface proteins before isolation and characterization is a valuable means of differentiating between plasma membrane and internal membrane proteins The results indicate that although the majority of cell-surface proteins can be identified using either of the approaches, others known to be important diagnostically and/or therapeutically would not be identified using either approach. The implication of this for the use of these techniques in the discovery of new leukocyte cell-surface proteins is discussed. PMID- 15531630 TI - High count rates with total internal reflection fluorescence correlation spectroscopy. PMID- 15531631 TI - Mechanically induced titin kinase activation studied by force-probe molecular dynamics simulations. AB - The conversion of mechanical stress into a biochemical signal in a muscle cell requires a force sensor. Titin kinase, the catalytic domain of the elastic muscle protein titin, has been suggested as a candidate. Its activation requires major conformational changes resulting in the exposure of its active site. Here, force probe molecular dynamics simulations were used to obtain insight into the tension induced activation mechanism. We find evidence for a sequential mechanically induced opening of the catalytic site without complete domain unfolding. Our results suggest the rupture of two terminal beta-sheets as the primary unfolding steps. The low force resistance of the C-terminal relative to the N-terminal beta sheet is attributed to their different geometry. A subsequent rearrangement of the autoinhibitory tail is seen to lead to the exposure of the active site, as is required for titin kinase activity. These results support the hypothesis of titin kinase as a force sensor. PMID- 15531632 TI - Pressure equilibrium and jump study on unfolding of 23-kDa protein from spinach photosystem II. AB - Pressure-induced unfolding of 23-kDa protein from spinach photosystem II has been systematically investigated at various experimental conditions. Thermodynamic equilibrium studies indicate that the protein is very sensitive to pressure. At 20 degrees C and pH 5.5, 23-kDa protein shows a reversible two-state unfolding transition under pressure with a midpoint near 160 MPa, which is much lower than most natural proteins studied to date. The free energy (DeltaG(u)) and volume change (DeltaV(u)) for the unfolding are 5.9 kcal/mol and -160 ml/mol, respectively. It was found that NaCl and sucrose significantly stabilize the protein from unfolding and the stabilization is associated not only with an increase in DeltaG(u) but also with a decrease in DeltaV(u). The pressure-jump studies of 23-kDa protein reveal a negative activation volume for unfolding ( 66.2 ml/mol) and a positive activation volume for refolding (84.1 ml/mol), indicating that, in terms of system volume, the protein transition state lies between the folded and unfolded states. Examination of the temperature effect on the unfolding kinetics indicates that the thermal expansibility of the transition state and the unfolded state of 23-kDa protein are closer to each other and they are larger than that of the native state. The diverse pressure-refolding pathways of 23-kDa protein in some conditions were revealed in pressure-jump kinetics. PMID- 15531633 TI - Multiphoton-FLIM quantification of the EGFP-mRFP1 FRET pair for localization of membrane receptor-kinase interactions. AB - We present an improved monomeric form of the red fluorescent protein, mRFP1, as the acceptor in biological fluorescence resonance energy transfer (FRET) experiments using the enhanced green fluorescent protein as donor. We find particular advantage in using this fluorophore pair for quantitative measurements of FRET using multiphoton fluorescence lifetime imaging microscopy (FLIM). The technique was exploited to demonstrate a novel receptor-kinase interaction between the chemokine receptor (CXCR4) and protein kinase C (PKC) alpha in carcinoma cells for both live- and fixed-cell experiments. The CXCR4-EGFP: PKCalpha-mRFP1 complex was found to be localized precisely to intracellular vesicles and cell protrusions when imaged by multiphoton fluorescence-FLIM. A comparison of the FRET efficiencies obtained using mRFP1-tagged regulatory domain or full-length PKCalpha as the acceptor revealed that PKCalpha, in the closed (inactive) form, is restrained from associating with the cytoplasmic portion of CXCR4. Live-cell FLIM experiments show that the assembly of this receptor:kinase complex is concomitant with the endocytosis process. This is confirmed by experimental evidence suggesting that the recycling of the CXCR4 receptor is increased on stimulation with phorbol ester and blocked on inhibition of PKC by bisindolylmaleimide. The EGFP-mRFP1 couple should be widely applicable, particularly to live-cell quantitative FRET assays. PMID- 15531634 TI - Modulation of mammalian inositol 1,4,5-trisphosphate receptor isoforms by calcium: a role of calcium sensor region. AB - In the accompanying article, we compared main functional properties of the three mammalian inositol 1,4,5-trisphosphate receptors (InsP3R) isoforms. In this article we focused on modulation of mammalian InsP3R isoforms by cytosolic Ca2+. We found that: 1), when recorded in the presence of 2 microM InsP3 and 0.5 mM ATP all three mammalian InsP3R isoforms display bell-shaped Ca2+ dependence in physiological range of Ca2+ concentrations (pCa 8-5); 2), in the same experimental conditions InsP3R3 is most sensitive to modulation by Ca2+ (peak at 107 nM Ca2+), followed by InsP3R2 (peak at 154 nM Ca2+), and then by InsP3R1 (peak at 257 nM Ca2+); 3), increase in ATP concentration to 5 mM had no significant effect of Ca2+ dependence of InsP3R1 and InsP3R2; 4), increase in ATP concentration to 5 mM converted Ca2+ dependence of InsP3R3 from "narrow" shape to "square" shape; 5), ATP-induced change in the shape of InsP3R3 Ca2+ dependence was mainly due to an >200-fold reduction in the apparent affinity of the Ca2+ inhibitory site; 6), the apparent Ca2+ affinity of the Ca2+ sensor region (Cas) determined in biochemical experiments is equal to 0.23 microM Ca2+ for RT1-Cas, 0.16 microM Ca2+ for RT2-Cas, and 0.10 microM Ca2+ for RT3-Cas; and 7), Ca2+ sensitivity of InsP3R1 and InsP3R3 isoforms recorded in the presence of 2 microM InsP3 and 0.5 mM ATP or 2 microM InsP3 and 5 mM ATP can be exchanged by swapping their Cas regions. Obtained results provide novel information about functional properties of mammalian InsP3R isoforms and support the importance of the Ca2+ sensor region (Cas) in determining the sensitivity of InsP3R isoforms to modulation by Ca2+. PMID- 15531635 TI - Exploring the energy landscape of GFP by single-molecule mechanical experiments. AB - We use single-molecule force spectroscopy to drive single GFP molecules from the native state through their complex energy landscape into the completely unfolded state. Unlike many smaller proteins, mechanical GFP unfolding proceeds by means of two subsequent intermediate states. The transition from the native state to the first intermediate state occurs near thermal equilibrium at approximately 35 pN and is characterized by detachment of a seven-residue N-terminal alpha-helix from the beta barrel. We measure the equilibrium free energy cost associated with this transition as 22 k(B)T. Detachment of this small alpha-helix completely destabilizes GFP thermodynamically even though the beta-barrel is still intact and can bear load. Mechanical stability of the protein on the millisecond timescale, however, is determined by the activation barrier of unfolding the beta barrel out of this thermodynamically unstable intermediate state. High bandwidth, time-resolved measurements of the cantilever relaxation phase upon unfolding of the beta-barrel revealed a second metastable mechanical intermediate with one complete beta-strand detached from the barrel. Quantitative analysis of force distributions and lifetimes lead to a detailed picture of the complex mechanical unfolding pathway through a rough energy landscape. PMID- 15531638 TI - Swimming of larval zebrafish: fin-axis coordination and implications for function and neural control. AB - Adult actinopterygian fishes typically perform steady forward swimming using either their pectoral fins or their body axis as the primary propulsor. In most species, when axial undulation is employed for swimming, the pectoral fins are tucked (i.e. adducted) against the body; conversely, when pectoral fins are beating, the body axis is held straight. In contrast to adults, larval fishes can combine their pectoral fin and body-axis movements during locomotion; however, little is known about how these locomotor modes are coordinated. With this study we provide a detailed analysis of the coordinated fin and axial movements during slow and fast swimming by examining forward locomotion in larval zebrafish (Danio rerio L.). In addition, we describe the musculature that powers pectoral fin movement in larval zebrafish and discuss its functional implications. As larvae, zebrafish alternate their pectoral fins during slow swimming (0.011+/-0.001 mm ms(-1)) in conjunction with axial undulations of the same frequency (18-28 Hz). During fast swimming (0.109+/-0.030 mm ms(-1); 36-67 Hz), the fins are tucked against the body and propulsion occurs by axial undulation alone. We show that during swimming, larval fishes can use a similar limb-axis coordination pattern to that of walking and running salamanders. We suggest that the fin-axis coordination observed in larval zebrafish may be attributed to a primitive neural circuit and that early terrestrial vertebrates may have gained the ability to coordinate limbs and lateral bending by retaining a larval central pattern generator for limb-axis coordination in the adult life history stage. PMID- 15531636 TI - RecA-dependent mutants in Escherichia coli reveal strategies to avoid chromosomal fragmentation. AB - RecA- and RecBC-catalyzed repair in eubacteria assembles chromosomes fragmented by double-strand breaks. We propose that recA mutants, being unable to repair fragmented chromosomes, depend on various strategies designed to avoid chromosomal fragmentation. To identify chromosomal fragmentation-avoidance strategies, we screened for Escherichia coli mutants synthetically inhibited in combination with recA inactivation by identifying clones unable to lose a plasmid carrying the recA(+) gene. Using this screen, we have isolated several RecA dependent mutants and assigned them to three distinct areas of metabolism. The tdk and rdgB mutants affect synthesis of DNA precursors. The fur, ubiE, and ubiH mutants are likely to have increased levels of reactive oxygen species. The seqA, topA mutants and an insertion in smtA perturbing the downstream mukFEB genes affect nucleoid administration. All isolated mutants show varying degree of SOS induction, indicating elevated levels of chromosomal lesions. As predicted, mutants in rdgB, seqA, smtA, topA, and fur show increased levels of chromosomal fragmentation in recBC mutant conditions. Future characterization of these RecA dependent mutants will define mechanisms of chromosomal fragmentation avoidance. PMID- 15531639 TI - Infrasound initiates directional fast-start escape responses in juvenile roach Rutilus rutilus. AB - Acoustic stimuli within the sonic range are effective triggers of C-type escape behaviours in fish. We have previously shown that fish have an acute sensitivity to infrasound also, with acceleration thresholds in the range of 10(-5) m s(-2). In addition, infrasound at high intensities around 10(-2) m s(-2) elicits strong and sustained avoidance responses in several fish species. In the present study, the possible triggering of C-escapes by infrasonic single-cycle vibrations was examined in juvenile roach Rutilus rutilus. The fish were accelerated in a controlled and quantifiable manner using a swing system. The applied stimuli simulated essential components of the accelerations that a small fish would encounter in the hydrodynamic flow field produced by a predatory fish. Typical C- and S-type escape responses were induced by accelerations within the infrasonic range with a threshold of 0.023 m s(-2) for an initial acceleration at 6.7 Hz. Response trajectories were on average in the same direction as the initial acceleration. Unexpectedly, startle behaviours mainly occurred in the trailing half of the test chamber, in which the fish were subjected to linear acceleration in combination with compression, i.e. the expected stimuli produced by an approaching predator. Very few responses were observed in the leading half of the test chamber, where the fish were subjected to acceleration and rarefaction, i.e. the stimuli expected from a suction type of predator. We conclude that particle acceleration is essential for the directionality of the startle response to infrasound, and that the response is triggered by the synergistic effects of acceleration and compression. PMID- 15531637 TI - Estrogen prevents bone loss through transforming growth factor beta signaling in T cells. AB - Estrogen (E) deficiency leads to an expansion of the pool of tumor necrosis factor (TNF)-producing T cells through an IFN-gamma-dependent pathway that results in increased levels of the osteoclastogenic cytokine TNF in the bone marrow. Disregulated IFN-gamma production is instrumental for the bone loss induced by ovariectomy (ovx), but the responsible mechanism is unknown. We now show that mice with T cell-specific blockade of type beta transforming growth factor (TGFbeta) signaling are completely insensitive to the bone-sparing effect of E. This phenotype results from a failure of E to repress IFN-gamma production, which, in turn, leads to increased T cell activation and T cell TNF production. Furthermore, ovx blunts TGFbeta levels in the bone marrow, and overexpression of TGFbeta in vivo prevents ovx-induced bone loss. These findings demonstrate that E prevents bone loss through a TGFbeta-dependent mechanism, and that TGFbeta signaling in T cells preserves bone homeostasis by blunting T cell activation. Thus, stimulation of TGFbeta production in the bone marrow is a critical "upstream" mechanism by which E prevents bone loss, and enhancement of TGFbeta levels in vivo may constitute a previously undescribed therapeutic approach for preventing bone loss. PMID- 15531640 TI - Allometry measurements from in situ video recordings can determine the size and swimming speeds of juvenile and adult squid Loligo opalescens (Cephalopoda: Myopsida). AB - Juvenile and adult Loligo opalescens Berry were video taped in Monterey Bay with the remotely operated vehicle (ROV) Ventana, captured with an otter trawl in Santa Monica Bay, California, and adults were taken from the Monterey Bay fishery. Behavioral observations were made over a 13 h period of video sequences. Allometry measurements were made on 157 squids ranging in size from 12 to 151 mm mantle length (ML). In addition to ML we measured the morphometric characters of fin length (FL), fin width (FW), mantle width (MW), eye diameter (ED), head width (HW), funnel aperture diameter (FA), fourth arm length (AL) and tentacle length (TL). Loligo opalescens changes shape with ontogeny due to negative allometric growth of ED, HW, TL, MW, FA and positive allometric growth of AL, FL and fin area. The allometry measurements were used to determine the size of juvenile squids video-taped in open water. A linear regression can predict dorsal ML in mm from a dimensionless ratio of ML upon ED (r2=0.857, P<0.001). Sizes and velocities of video-taped animals were estimated from 26 video sequences ranging from <1.0 to 8 s. The average velocity for squids ranging from 12-116 mm ML was 0.21 m s(-1) and the maximum velocity was 1.60 m s(-1) (116 mm ML). Allometric measurements can provide scale for 2-dimensional images in order to estimate size, velocity and age of animals. PMID- 15531641 TI - Variation of crustacean hyperglycemic hormone (cHH) level in the eyestalk and haemolymph of the shrimp Palaemon elegans following stress. AB - This study investigates (by means of bioassays and ELISA using an antibody against recombinant cHH) the variation of cHH levels in the eyestalks and haemolymph of Palaemon elegans (Decapoda, Caridea) following exposure to various stresses (heavy metals and lipopolysaccharide), and correlates them with the variation in amount and time course of blood glucose. The dose-relationship between exposure to copper and quick release of cHH from the eyestalk into haemolymph was confirmed by variation of blood glucose with a dose-related hyperglycaemia, that peaked 2 h after immersion in contaminated seawater. Animals exposed to a sublethal concentration of mercury showed the same dose relation between toxicant, release of cHH from the eyestalk, increment of circulating hormone level and subsequent hyperglycaemia as observed for copper contamination. It is of note that although the highest lethal mercury concentration induced the release of cHH from the eyestalk into the haemolymph, it was not followed by a significant variation of blood glucose. Step doses of a bacterial contaminant [such as lipopolysaccharide (LPS) from E. coli injected into shrimps] confirmed the dose-relationship and convergent chain of events that bring about hyperglycaemia. These are the first data that relate the release of cHH from the eyestalk, the circulating hormone level and the consequent glycaemic response to stress. Moreover, they confirm the dose-related pathway that leads to variation of blood glucose as a quantitative biomarker of environmental quality, even at sublethal toxicant concentrations. PMID- 15531642 TI - Biomechanical and energetic determinants of the walk-trot transition in horses. AB - We studied nine adult horses spanning an eightfold range in body mass (M(b)) (90 720 kg) and a twofold range in leg length (L) (0.7-1.4 m). We measured the horses' walk-trot transition speeds using step-wise speed increments as they locomoted on a motorized treadmill. We then measured their rates of oxygen consumption over a wide range of walking and trotting speeds. We interpreted the transition speed results using a simple inverted-pendulum model of walking in which gravity provides the centripetal force necessary to keep the leg in contact with the ground. By studying a large size range of horses, we were naturally able to vary the absolute walking speed that would produce the same ratio of centripetal to gravitational forces. This ratio, (M(b)v2/L)/(M(b)g), reduces to the dimensionless Froude number (v2/gL), where v is forward speed, L is leg length and g is gravitational acceleration. We found that the absolute walk-trot transition speed increased with size from 1.6 to 2.3 m s(-1), but it occurred at nearly the same Froude number (0.35). In addition, horses spontaneously switched between gaits in a narrow range of speeds that corresponded to the metabolically optimal transition speed. These results support the hypotheses that the walk-trot transition is triggered by inverted-pendulum dynamics and occurs at the speed that maximizes metabolic economy. PMID- 15531643 TI - Effect of acute heat stress on rat adrenal glands: a morphological and stereological study. AB - The morphological and stereological structure of rat adrenal gland was analysed by light microscopy after an acute (60 min) exposure to high ambient temperature (38 degrees C). A significant increase in plasma corticotrophin (ACTH) and serum corticosterone (CORT) concentrations was observed, confirming that acute heat exposure has a strong stressful effect. Under these conditions the adrenal gland mass and volume were decreased, probably as the consequence of adrenal cortex reduction, especially that of the zona fasciculata (ZF). Histological examination revealed that many ZF cells were deprived of lipid droplets. Fibrosis was observed in all parts of the adrenal gland, both cortex and medulla, of heat stressed animals. Mitotic figures were absent in cortical cells after heat exposure, but there were no differences in ZF and zona reticularis (ZR) small blood vessels compared to nonstressed controls. PMID- 15531644 TI - Heat in evolution's kitchen: evolutionary perspectives on the functions and origin of the facial pit of pitvipers (Viperidae: Crotalinae). AB - Pitvipers (Viperidae: Crotalinae) possess thermal radiation receptors, the facial pits, which allow them to detect modest temperature fluctuations within their environments. It was previously thought that these organs were used solely to aid in prey acquisition, but recent findings demonstrated that western diamondback rattlesnakes (Crotalus atrox) use them to direct behavioral thermoregulation, suggesting that facial pits might be general purpose organs used to drive a suite of behaviors. To investigate this further, we conducted a phylogenetic survey of viperine thermoregulatory behavior cued by thermal radiation. We assessed this behavior in 12 pitviper species, representing key nodes in the evolution of pitvipers and a broad range of thermal environments, and a single species of true viper (Viperidae: Viperinae), a closely related subfamily of snakes that lack facial pits but possess a putative thermal radiation receptor. All pitviper species were able to rely on their facial pits to direct thermoregulatory movements, while the true viper was unable to do so. Our results suggest that thermoregulatory behavior cued by thermal radiation is a universal role of facial pits and probably represents an ancestral trait among pitvipers. Further, they establish behavioral thermoregulation as a plausible hypothesis explaining the evolutionary origin of the facial pit. PMID- 15531645 TI - Temperature and the expression of myogenic regulatory factors (MRFs) and myosin heavy chain isoforms during embryogenesis in the common carp Cyprinus carpio L. AB - Embryos of the common carp, Cyprinus carpio L., were reared from fertilization of the eggs to inflation of the swim bladder in the larval stage at 18 and 25 degrees C. cRNA probes were used to detect transcripts of the myogenic regulatory factors MyoD, Myf-5 and myogenin, and five myosin heavy chain (MyHC) isoforms during development. The genes encoding Myf-5 and MyoD were switched on first in the unsegmented mesoderm, followed by myogenin as the somites developed. Myf-5 and MyoD transcripts were initially limited to the adaxial cells, but Myf-5 expression spread laterally into the presomitic mesoderm before somite formation. Two distinct bands of staining could be seen corresponding to the cellular fields of the forming somites, but as each furrow delineated, Myf-5 mRNA levels declined. Upon somite formation, MyoD expression spread laterally to encompass the full somite width. Expression of the myogenin gene was also switched on during somite formation, and expression of both transcripts persisted until the somites became chevron-shaped. Expression of MyoD was then downregulated shortly before myogenin. The expression patterns of the carp myogenic regulatory factor (MRF) genes most-closely resembled that seen in the zebrafish rather than the rainbow trout (where expression of MyoD remains restricted to the adaxial domain of the somite for a prolonged period) or the herring (where expression of MyoD persists longer than that of myogenin). Expression of two embryonic forms of MyHC began simultaneously at the 25-30 somite stage and continued until approximately two weeks post-hatch. However, the three adult isoforms of fast muscle MyHC were not detected in any stage examined, emphasizing a developmental gap that must be filled by other, as yet uncharacterised, MyHC isoform(s). No differences in the timing of expression of any mRNA transcripts were seen between temperature groups. A phylogenetic analysis of the MRFs was conducted using all available full-length amino acid sequences. A neighbour-joining tree indicated that all four members evolved from a common ancestral gene, which first duplicated into two lineages, each of which underwent a further duplication to produce Myf-5 and MyoD, and myogenin and MRF4. Parologous copies of MyoD from trout and Xenopus clustered closely together within clades, indicating recent duplications. By contrast, MyoD paralogues from gilthead seabream were more divergent, indicating a more-ancient duplication. PMID- 15531646 TI - Spatial vision in the echinoid genus Echinometra. AB - Although eyes are generally considered necessary for image resolution, a diffuse photoreceptive system with directional sensitivity may also have this ability. Two species of the echinoid genus Echinometra were tested for spatial vision by examining their ability to locate and move towards targets of different sizes. The echinoids were significantly oriented (P<0.0001) towards a target with an angular width of 33 degrees (0.3 sr) but were not oriented to targets with angular widths of 26 degrees and 16 degrees . This ability is probably due to the blocking of off-angle light by the spines, which have approximately the correct spacing for the observed resolution. Spatial vision is advantageous for echinoids of this genus because they leave and return to small dark shelters. This first demonstration of spatial vision in an echinoderm sheds further light on the complex optical structures and photobehaviors found in this phylum. PMID- 15531647 TI - The relationship between wingbeat kinematics and vortex wake of a thrush nightingale. AB - The wingbeat kinematics of a thrush nightingale Luscinia luscinia were measured for steady flight in a wind tunnel over a range of flight speeds (5-10 m s(-1)), and the results are interpreted and discussed in the context of a detailed, previously published, wake analysis of the same bird. Neither the wingbeat frequency nor wingbeat amplitude change significantly over the investigated speed range and consequently dimensionless measures that compare timescales of flapping vs. timescales due to the mean flow vary in direct proportion to the mean flow itself, with no constant or slowly varying intervals. The only significant kinematic variations come from changes in the upstroke timing (downstroke fraction) and the upstroke wing folding (span ratio), consistent with the gradual variations, primarily in the upstroke wake, previously reported. The relationship between measured wake geometry and wingbeat kinematics can be qualitatively explained by presumed self-induced convection and deformation of the wake between its initial formation and later measurement, and varies in a predictable way with flight speed. Although coarse details of the wake geometry accord well with the kinematic measurements, there is no simple explanation based on these observed kinematics alone that accounts for the measured asymmetries of circulation magnitude in starting and stopping vortex structures. More complex interactions between the wake and wings and/or body are implied. PMID- 15531648 TI - The effect of advance ratio on the aerodynamics of revolving wings. AB - Recent studies have demonstrated that a quasi-steady model closely matches the instantaneous force produced by an insect wing during hovering flight. It is not clear, however, if such methods extend to forward flight. In this study we use a dynamically scaled robotic model of the fruit fly Drosophila melanogaster to investigate the forces produced by a wing revolving at constant angular velocity while simultaneously translating at velocities appropriate for forward flight. Because the forward and angular velocities were constant wing inertia was negligible, and the measured forces can be attributed to fluid dynamic phenomena. The combined forward and revolving motions of the wing produce a time-dependent free-stream velocity profile, which suggests that added mass forces make a contribution to the measured forces. We find that the forces due added mass make a small, but measurable, component of the total force and are in excellent agreement with theoretical values. Lift and drag coefficients are calculated from the force traces after subtracting the contributions due to added mass. The lift and drag coefficients, for fixed angle of attack, are not constant for non-zero advance ratios, but rather vary in magnitude throughout the stroke. This observation implies that modifications of the quasi-steady model are required in order to predict accurately the instantaneous forces produced during forward flight. We show that the dependence of the lift and drag coefficients upon advance ratio and stroke position can be characterized effectively in terms of the tip velocity ratio--the ratio of the chordwise components of flow velocity at the wing tip due to translation and revolution. On this basis we develop a modified quasi-steady model that can account for the varying magnitudes of the lift and drag coefficients. Our model may also resolve discrepancies in past measurements of wing performance based on translational and revolving motion. PMID- 15531649 TI - Rumen metabolites serve ticks to exploit large mammals. AB - Hard ticks spend most of their life isolated from passing vertebrates but require a blood meal to proceed to the next life stage (larva, nymph or adult). These opportunist ectoparasites must be capable of anticipating signals that render suitable hosts apparent. Large ungulates that tolerate a high ectoparasite burden are the favoured hosts of adult hard ticks. Ruminants, comprising the majority of ungulate species, must regularly eruct gases from the foregut to relieve excess pressure and maintain a chemical equilibrium. Through eructations from individuals, and particularly herds, ruminants inadvertently signal their presence to hard ticks. Here, we report that all adult hard tick species we tested are attracted to cud and demonstrate that these acarines possess olfactory receptor cells for the carboxylic acid, phenol and indole end-products of the rumen bioreactor. Compounds from each of these classes of volatiles attract ticks on their own, and mixtures of these volatiles based on rumen composition also attract. Appetence for rumen metabolites represents a fundamental resource tracking adaptation by hard ticks for large roaming mammals. PMID- 15531650 TI - Metabolic power of European starlings Sturnus vulgaris during flight in a wind tunnel, estimated from heat transfer modelling, doubly labelled water and mask respirometry. AB - It is technically demanding to measure the energetic cost of animal flight. Each of the previously available techniques has some disadvantage as well advantages. We compared measurements of the energetic cost of flight in a wind tunnel by four European starlings Sturnus vulgaris made using three independent techniques: heat transfer modelling, doubly labelled water (DLW) and mask respirometry. We based our heat transfer model on thermal images of the surface temperature of the birds and air flow past the body and wings calculated from wing beat kinematics. Metabolic power was not sensitive to uncertainty in the value of efficiency when estimated from heat transfer modelling. A change in the assumed value of whole animal efficiency from 0.19 to 0.07 (the range of estimates in previous studies) only altered metabolic power predicted from heat transfer modelling by 13%. The same change in the assumed value of efficiency would cause a 2.7-fold change in metabolic power if it were predicted from mechanical power. Metabolic power did not differ significantly between measurements made using the three techniques when we assumed an efficiency in the range 0.11-0.19, although the DLW results appeared to form a U-shaped power-speed curve while the heat transfer model and respirometry results increased linearly with speed. This is the first time that techniques for determining metabolic power have been compared using data from the same birds flying under the same conditions. Our data provide reassurance that all the techniques produce similar results and suggest that heat transfer modelling may be a useful method for estimating metabolic rate. PMID- 15531651 TI - Dragonfly flight: free-flight and tethered flow visualizations reveal a diverse array of unsteady lift-generating mechanisms, controlled primarily via angle of attack. AB - Here we show, by qualitative free- and tethered-flight flow visualization, that dragonflies fly by using unsteady aerodynamic mechanisms to generate high-lift, leading-edge vortices. In normal free flight, dragonflies use counterstroking kinematics, with a leading-edge vortex (LEV) on the forewing downstroke, attached flow on the forewing upstroke, and attached flow on the hindwing throughout. Accelerating dragonflies switch to in-phase wing-beats with highly separated downstroke flows, with a single LEV attached across both the fore- and hindwings. We use smoke visualizations to distinguish between the three simplest local analytical solutions of the Navier-Stokes equations yielding flow separation resulting in a LEV. The LEV is an open U-shaped separation, continuous across the thorax, running parallel to the wing leading edge and inflecting at the tips to form wingtip vortices. Air spirals in to a free-slip critical point over the centreline as the LEV grows. Spanwise flow is not a dominant feature of the flow field--spanwise flows sometimes run from wingtip to centreline, or vice versa- depending on the degree of sideslip. LEV formation always coincides with rapid increases in angle of attack, and the smoke visualizations clearly show the formation of LEVs whenever a rapid increase in angle of attack occurs. There is no discrete starting vortex. Instead, a shear layer forms behind the trailing edge whenever the wing is at a non-zero angle of attack, and rolls up, under Kelvin-Helmholtz instability, into a series of transverse vortices with circulation of opposite sign to the circulation around the wing and LEV. The flow fields produced by dragonflies differ qualitatively from those published for mechanical models of dragonflies, fruitflies and hawkmoths, which preclude natural wing interactions. However, controlled parametric experiments show that, provided the Strouhal number is appropriate and the natural interaction between left and right wings can occur, even a simple plunging plate can reproduce the detailed features of the flow seen in dragonflies. In our models, and in dragonflies, it appears that stability of the LEV is achieved by a general mechanism whereby flapping kinematics are configured so that a LEV would be expected to form naturally over the wing and remain attached for the duration of the stroke. However, the actual formation and shedding of the LEV is controlled by wing angle of attack, which dragonflies can vary through both extremes, from zero up to a range that leads to immediate flow separation at any time during a wing stroke. PMID- 15531652 TI - The biodynamics of arboreal locomotion: the effects of substrate diameter on locomotor kinetics in the gray short-tailed opossum (Monodelphis domestica). AB - Effects of substrate diameter on locomotor biodynamics were studied in the gray short-tailed opossum (Monodelphis domestica). Two horizontal substrates were used: a flat 'terrestrial' trackway with a force platform integrated into the surface and a cylindrical 'arboreal' trackway (20.3 mm diameter) with a force transducer instrumented region. On both terrestrial and arboreal substrates, fore limbs exhibited higher vertical impulse and peak vertical force than hind limbs. Although vertical limb impulses were lower on the terrestrial substrate than on the arboreal support, this was probably due to speed effects because the opossums refused to move as quickly on the arboreal trackway. Vertical impulse decreased significantly faster with speed on the arboreal substrate because most of these trials were relatively slow, and stance duration decreased with speed more rapidly at these lower speeds. While braking and propulsive roles were more segregated between limbs on the terrestrial trackway, fore limbs were dominant both in braking and in propulsion on the arboreal trackway. Both fore and hind limbs exerted equivalently strong, medially directed limb forces on the arboreal trackway and laterally directed limb forces on the terrestrial trackway. We propose that the modifications in substrate reaction force on the arboreal trackway are due to the differential placement of the limbs about the dorsolateral aspect of the branch. Specifically, the pes typically made contact with the branch lower and more laterally than the manus, which may explain the significantly lower required coefficient of friction in the fore limbs relative to the hind limbs. PMID- 15531654 TI - Saturated fat prevents coronary artery disease? An American paradox. PMID- 15531653 TI - Futile short-patch DNA base excision repair of adenine:8-oxoguanine mispair. AB - 8-oxo-7, 8-dihydrodeoxyguanosine (8-oxo-dG), one of the representative oxidative DNA lesions, frequently mispairs with the incoming dAMP during mammalian DNA replication. Mispaired dA is removed by post-replicative base excision repair (BER) initiated by adenine DNA glycosylase, MYH, creating an apurinic (AP) site. The subsequent mechanism ensuring a dC:8-oxo-dG pair, a substrate for 8 oxoguanine DNA glycosylase (OGG1), remains to be elucidated. At the nucleotide insertion step, none of the mammalian DNA polymerases examined exclusively inserted dC opposite 8-oxo-dG that was located in a gap. AP endonuclease 1, which possesses 3'-->5' exonuclease activity and potentially serves as a proofreader, did not discriminate dA from dC that was located opposite 8-oxo-dG. However, human DNA ligases I and III joined 3'-dA terminus much more efficiently than 3' dC terminus when paired to 8-oxo-dG. In reconstituted short-patch BER, repair products contained only dA opposite 8-oxo-dG. These results indicate that human DNA ligases discriminate dC from dA and that MYH-initiated short-patch BER is futile and hence this BER must proceed to long-patch repair, even if it is initiated as short-patch repair, through strand displacement synthesis from the ligation-resistant dC terminus to generate the OGG1 substrate, dC:8-oxo-dG pair. PMID- 15531655 TI - New approaches to iron fortification: role of bioavailability studies. PMID- 15531656 TI - Human aflatoxicosis in developing countries: a review of toxicology, exposure, potential health consequences, and interventions. AB - Aflatoxins are well recognized as a cause of liver cancer, but they have additional important toxic effects. In farm and laboratory animals, chronic exposure to aflatoxins compromises immunity and interferes with protein metabolism and multiple micronutrients that are critical to health. These effects have not been widely studied in humans, but the available information indicates that at least some of the effects observed in animals also occur in humans. The prevalence and level of human exposure to aflatoxins on a global scale have been reviewed, and the resulting conclusion was that approximately 4.5 billion persons living in developing countries are chronically exposed to largely uncontrolled amounts of the toxin. A limited amount of information shows that, at least in those locations where it has been studied, the existing aflatoxin exposure results in changes in nutrition and immunity. The aflatoxin exposure and the toxic affects of aflatoxins on immunity and nutrition combine to negatively affect health factors (including HIV infection) that account for >40% of the burden of disease in developing countries where a short lifespan is prevalent. Food systems and economics render developed-country approaches to the management of aflatoxins impractical in developing-country settings, but the strategy of using food additives to protect farm animals from the toxin may also provide effective and economical new approaches to protecting human populations. PMID- 15531657 TI - Will mandatory folic acid fortification prevent or promote cancer? AB - An overwhelming body of evidence for a protective effect of periconceptional folic acid supplementation against neural tube defects (NTDs) led to mandatory folic acid fortification in the United States. The effectiveness of folic acid fortification in improving folate status has already been shown to be quite striking, with a dramatic increase in blood measurements of folate in the United States. Preliminary reports also suggest a significant reduction ( approximately 15-50%) in NTDs in the United States. The success of folic acid fortification in improving folate status and in reducing NTD rates is truly a public health triumph and provides a paradigm of collaboration between science and public health policy. Although folic acid is generally regarded as safe, there continues to be concern that folic acid fortification may have adverse effects in subpopulation groups not originally targeted for fortification. In this regard, an emerging body of evidence suggests that folic acid supplementation may enhance the development and progression of already existing, undiagnosed premalignant and malignant lesions. Over the past few years, the US population has been exposed to a significant increase in folate intake, for which essentially no data on safety exist. The potential cancer-promoting effect of folic acid supplementation needs to be considered in carefully monitoring the long-term effect of folic acid fortification on the vast majority of the US population, who are not at risk of NTDs. PMID- 15531658 TI - Appropriate body mass index and waist circumference cutoffs for categorization of overweight and central adiposity among Chinese adults. AB - BACKGROUND: Recent data suggest that current overweight and central adiposity guidelines based on Western populations are not appropriate for Asian populations. The published data among Chinese are insufficient to address this issue. OBJECTIVE: We aimed to identify cutoffs for body mass index (BMI; in kg/m(2)) and waist circumference that confer increased risk of cardiovascular disease in Chinese adults as would be consistent with overweight and central adiposity. DESIGN: A nationally representative, cross-sectional sample of 15 239 Chinese adults aged 35-74 y was studied. RESULTS: Mean blood pressure, total cholesterol, LDL-cholesterol, triacylglycerol, and glucose values were incrementally higher and mean HDL-cholesterol values were incrementally lower with each unit increase in BMI and waist circumference in both men and women. Both the point at which sensitivity equaled specificity and the shortest distance in the receiver operating characteristic curves for hypertension, dyslipidemia, diabetes, or >/=2 of these risk factors suggested a BMI cutoff of 24 and a waist circumference cutoff of 80 cm for both men and women. CONCLUSIONS: Lower cutoffs for BMI and waist circumference are needed in the identification of Chinese patients at high risk of cardiovascular disease. PMID- 15531659 TI - Fish oil and antioxidants alter the composition and function of circulating mononuclear cells in Crohn disease. AB - BACKGROUND: Crohn disease (CD) is associated with osteoporosis and other extraintestinal manifestations that might be mediated by cytokines from circulating (peripheral blood) mononuclear cells (PBMCs). Fish oil rich in eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) reduces disease activity in patients with CD with raised laboratory markers of inflammation and in healthy subjects alters PBMC function. OBJECTIVE: We investigated the effect of fish oil plus antioxidants on cytokine production by PBMCs from patients with CD with raised C-reactive protein concentrations (>/=6.9 mg/L) or erythrocyte sedimentation rates (>/=18 mm/h). DESIGN: A randomized placebo-controlled trial of fish oil (2.7 g EPA and DHA/d; n = 31) or placebo (olive oil; n = 31) for 24 wk was conducted in patients with CD. The fish-oil group additionally received an antioxidant preparation (vitamins A, C, and E and selenium). Exclusion criteria included corticosteroid use. Fatty acid composition was measured by gas chromatography. Production of tumor necrosis factor alpha, interferon gamma (IFN gamma), and prostaglandin E(2) (PGE(2)) was measured by enzyme-linked immunosorbent assays after stimulation with mitogen and endotoxin (lipopolysaccharide). RESULTS: Fish-oil plus antioxidant dietary supplementation was associated with higher EPA and DHA incorporation into PBMCs (P < 0.001) and lower arachidonic acid (P = 0.006) and lower production of IFN-gamma by mitogen stimulated PBMCs (P = 0.012) and of PGE(2) by lipopolysaccharide-stimulated PBMCs (P = 0.047). CONCLUSION: Dietary supplementation with fish oil plus antioxidants is associated with modified PBMC composition and lower production of PGE(2) and IFN-gamma by circulating monocytes or macrophages. The response of extraintestinal manifestations of CD should be investigated in a randomized controlled trial. PMID- 15531660 TI - Contribution of postprandial lipemia to the dietary fat-mediated changes in endogenous lipoprotein-cholesterol concentrations in humans. AB - BACKGROUND: Dietary fats alter LDL and HDL cholesterol while serving as precursors of postprandial triacylglycerol-rich lipoproteins (TRLs). OBJECTIVE: We hypothesized that the saturated fatty acid (SFA)-mediated increase and the polyunsaturated fatty acid (PUFA)-mediated decrease in endogenous lipoprotein cholesterol are promoted by postprandial TRLs. DESIGN: We performed a 16-d crossover diet study to examine the effect of PUFA-rich [ratio of PUFAs to SFAs (P:S) = 2.0] and SFA-rich (P:S = 0.25) diets on fasting and postprandial plasma lipid and lipoprotein-cholesterol concentrations in 16 normolipidemic subjects. RESULTS: Fasting plasma cholesterol decreased significantly after a PUFA-rich diet because of a decrease in LDL (-12.3%; P < 0.05) and HDL (-3.8%; NS), but did not change after an SFA-rich diet. The appearance of postprandial TRLs in plasma at 4 h was linked to a significant lowering of both LDL (-7.4%) and HDL (-4.8%) after a PUFA-rich diet; no such effect was observed after the SFA-rich diet. At 7 h, LDL and HDL cholesterol returned to near fasting concentrations without postprandial TRL accumulation after a PUFA-rich diet but with a significant postprandial TRL accumulation after an SFA-rich diet. Thus, the in vivo postprandial clearance of cholesterol in LDL+HDL was greater after a PUFA-rich diet than after an SFA-rich diet. The appearance of postprandial TRLs in plasma increased the cholesteryl ester transfer protein-mediated transfer of cholesteryl ester from LDL+HDL to TRLs in vitro without a significant influence from dietary fat. CONCLUSION: Dietary fat-mediated alterations in the rate of hepatic removal of postprandial TRLs, which carry cholesterol accepted from LDL+HDL via cholesteryl ester transfer protein in vivo, may contribute to the dietary fat mediated change in endogenous lipoprotein cholesterol. PMID- 15531661 TI - Plant sterols and endurance training combine to favorably alter plasma lipid profiles in previously sedentary hypercholesterolemic adults after 8 wk. AB - BACKGROUND: Plant sterol supplementation was shown to reduce total and LDL cholesterol concentrations, whereas endurance training was shown to increase HDL cholesterol concentrations and decrease triacylglycerol concentrations. OBJECTIVE: The objective was to examine the effect of plant sterols, endurance training, and the combination of plant sterols and endurance training on plasma lipid and lipoprotein cholesterol concentrations, sterol concentrations, and cholesterol precursor concentrations in previously sedentary hypercholesterolemic adults. DESIGN: In an 8-wk, placebo-controlled, parallel-arm clinical trial, 84 subjects were randomly assigned to receive 1 of 4 interventions: 1) combination of sterols and exercise, 2) exercise, 3) sterols, or 4) control treatment. RESULTS: Sterol supplementation significantly (P < 0.01) decreased total cholesterol concentrations by 8.2% from baseline. In addition, sterols significantly (P < 0.01) lowered absolute LDL-cholesterol concentrations after treatment but had no effect on the percentage change from the beginning to the end of the trial. Exercise significantly (P < 0.01) increased HDL-cholesterol concentrations by 7.5% and decreased triacylglycerol concentrations by 13.3% from baseline. Moreover, sterol supplementation significantly (P < 0.05) increased lathosterol, campesterol, and beta-sitosterol concentrations after treatment. Exercise significantly (P < 0.01) decreased percentage of body fat by 3.9% from the beginning to the end of the trial. CONCLUSIONS: In comparison with plant sterols or exercise alone, the combination of plant sterols and exercise yields the most beneficial alterations in lipid profiles. Implementation of such a combination therapy could improve lipid profiles in those at risk of coronary artery disease. PMID- 15531662 TI - Docosahexaenoic acid concentrations are higher in women than in men because of estrogenic effects. AB - BACKGROUND: During pregnancy there is a high demand for docosahexaenoic acid (DHA), which is needed for formation of the fetal brain. Women who do not consume marine foods must synthesize DHA from fatty acid precursors in vegetable foods. OBJECTIVE: We studied sex differences in DHA status and the role of sex hormones. DESIGN: First, DHA status was compared between 72 male and 103 female healthy volunteers who ate the same rigidly controlled diets. Second, the effects of sex hormones were studied in 56 male-to-female transsexual subjects, who were treated with cyproterone acetate alone or randomly assigned to receive oral ethinyl estradiol or transdermal 17beta-estradiol combined with cyproterone acetate, and in 61 female-to-male transsexual subjects, who were treated with testosterone esters or randomly assigned for treatment with the aromatase inhibitor anastrozole or placebo in addition to the testosterone regimen. RESULTS: The proportion of DHA was 15 +/- 4% (x +/- SEM; P < 0.0005) higher in the women than in the men. Among the women, those taking oral contraceptives had 10 +/- 4% (P = 0.08) higher DHA concentrations than did those not taking oral contraceptives. Administration of oral ethinyl estradiol, but not transdermal 17beta-estradiol, increased DHA by 42 +/- 8% (P < 0.0005), whereas the antiandrogen cyproterone acetate did not affect DHA. Parenteral testosterone decreased DHA by 22 +/- 4% (P < 0.0005) in female-to-male transsexual subjects. Anastrozole decreased estradiol concentrations significantly and DHA concentrations nonsignificantly (9 +/- 6%; P = 0.09). CONCLUSION: Estrogens cause higher DHA concentrations in women than in men, probably by upregulating synthesis of DHA from vegetable precursors. PMID- 15531663 TI - Dietary fats, carbohydrate, and progression of coronary atherosclerosis in postmenopausal women. AB - BACKGROUND: The influence of diet on atherosclerotic progression is not well established, particularly in postmenopausal women, in whom risk factors for progression may differ from those for men. OBJECTIVE: The objective was to investigate associations between dietary macronutrients and progression of coronary atherosclerosis among postmenopausal women. DESIGN: Quantitative coronary angiography was performed at baseline and after a mean follow-up of 3.1 y in 2243 coronary segments in 235 postmenopausal women with established coronary heart disease. Usual dietary intake was assessed at baseline. RESULTS: The mean (+/-SD) total fat intake was 25 +/- 6% of energy. In multivariate analyses, a higher saturated fat intake was associated with a smaller decline in mean minimal coronary diameter (P = 0.001) and less progression of coronary stenosis (P = 0.002) during follow-up. Compared with a 0.22-mm decline in the lowest quartile of intake, there was a 0.10-mm decline in the second quartile (P = 0.002), a 0.07 mm decline in the third quartile (P = 0.002), and no decline in the fourth quartile (P < 0.001); P for trend = 0.001. This inverse association was more pronounced among women with lower monounsaturated fat (P for interaction = 0.04) and higher carbohydrate (P for interaction = 0.004) intakes and possibly lower total fat intake (P for interaction = 0.09). Carbohydrate intake was positively associated with atherosclerotic progression (P = 0.001), particularly when the glycemic index was high. Polyunsaturated fat intake was positively associated with progression when replacing other fats (P = 0.04) but not when replacing carbohydrate or protein. Monounsaturated and total fat intakes were not associated with progression. CONCLUSIONS: In postmenopausal women with relatively low total fat intake, a greater saturated fat intake is associated with less progression of coronary atherosclerosis, whereas carbohydrate intake is associated with a greater progression. PMID- 15531664 TI - Diets containing barley significantly reduce lipids in mildly hypercholesterolemic men and women. AB - BACKGROUND: Barley has high amounts of soluble fiber but is not extensively consumed in the US diet. OBJECTIVE: This study investigated whether consumption of barley would reduce cardiovascular disease risk factors comparably with that of other sources of soluble fiber. DESIGN: Mildly hypercholesterolemic subjects (9 postmenopausal women, 9 premenopausal women, and 7 men) consumed controlled American Heart Association Step 1 diets for 17 wk. After a 2-wk adaptation period, whole-grain foods containing 0, 3, or 6 g beta-glucan/d from barley were included in the Step 1 diet menus. Diets were consumed for 5 wk each and were fed in a Latin-square design. Fasting blood samples were collected twice weekly. RESULTS: Total cholesterol was significantly lower when the diet contained 3 or 6 g beta-glucan/d from barley than when it contained no beta-glucan; the greatest change occurred in the men and postmenopausal women. HDL and triacylglycerol concentrations did not differ with the 3 amounts of dietary beta-glucan. Large LDL and small VLDL fractions and mean LDL particle size significantly decreased when whole grains were incorporated into the 3 diets. Large LDL and large and intermediate HDL fractions were significantly higher, mean LDL particle size was significantly greater, and intermediate VLDL fractions were significantly lower in the postmenopausal women than in the other 2 groups. A group-by-diet interaction effect was observed on LDL fractions and small LDL particle size. CONCLUSION: The addition of barley to a healthy diet may be effective in lowering total and LDL cholesterol in both men and women. PMID- 15531665 TI - Does supplemental vitamin C increase cardiovascular disease risk in women with diabetes? AB - BACKGROUND: Vitamin C acts as a potent antioxidant; however, it can also be a prooxidant and glycate protein under certain circumstances in vitro. These observations led us to hypothesize that a high intake of vitamin C in diabetic persons might promote atherosclerosis. OBJECTIVE: The objective was to examine the relation between vitamin C intake and mortality from cardiovascular disease. DESIGN: We studied the relation between vitamin C intake and mortality from total cardiovascular disease (n = 281), coronary artery disease (n = 175), and stroke (n = 57) in 1923 postmenopausal women who reported being diabetic at baseline. Diet was assessed with a food-frequency questionnaire at baseline, and subjects initially free of coronary artery disease were prospectively followed for 15 y. RESULTS: After adjustment for cardiovascular disease risk factors, type of diabetes medication used, duration of diabetes, and intakes of folate, vitamin E, and beta-carotene, the adjusted relative risks of total cardiovascular disease mortality were 1.0, 0.97, 1.11, 1.47, and 1.84 (P for trend < 0.01) across quintiles of total vitamin C intake from food and supplements. Adjusted relative risks of coronary artery disease were 1.0, 0.81, 0.99, 1.26, and 1.91 (P for trend = 0.01) and of stroke were 1.0, 0.52, 1.23, 2.22, and 2.57 (P for trend < 0.01). When dietary and supplemental vitamin C were analyzed separately, only supplemental vitamin C showed a positive association with mortality endpoints. Vitamin C intake was unrelated to mortality from cardiovascular disease in the nondiabetic subjects at baseline. CONCLUSION: A high vitamin C intake from supplements is associated with an increased risk of cardiovascular disease mortality in postmenopausal women with diabetes. PMID- 15531666 TI - Nutritional status is an important predictor of diaphragm strength in young patients with cystic fibrosis. AB - BACKGROUND: The effect of nutritional status and lung disease progression on diaphragm strength in young patients with cystic fibrosis remains unclear. OBJECTIVE: The aim of this study was to investigate the effect of nutritional status and airway obstruction on diaphragm strength. DESIGN: Twitch transdiaphragmatic pressure (Tw Pdi) obtained by bilateral anterior magnetic phrenic nerve stimulation, body mass index (BMI) z score, fat mass, fat-free mass (FFM), arm muscle circumference (AMC), forced expiratory volume in 1 s (FEV(1)), and functional residual capacity (FRC) were measured in 20 patients aged 15.1 +/- 2.8 y (x +/- SD). Values were expressed as a percentage of predicted values. RESULTS: Mean (+/-SD) Tw Pdi was 24.3 +/- 5.5 cm H(2)O. Univariate regression analysis showed positive correlations between Tw Pdi and nutrition scores (BMI z score: r = 0.63, P = 0.003; FFM: r = 0.47, P = 0.04; AMC: r = 0.45, P = 0.04), airway obstruction (FEV(1): r = 0.68, P = 0.001), and arterial oxygen partial pressure (r = 0.68, P = 0.001). Negative correlations were observed between Tw Pdi and dynamic hyperinflation (FRC: r = -0.65, P = 0.005) and arterial carbon dioxide pressure (r = -0.50, P = 0.03). Furthermore, stepwise regression analysis showed that Tw Pdi correlated with BMI z score (r = 0.75, P = 0.0002) and FEV(1) (r = 0.69, P = 0.001). CONCLUSIONS: Diaphragm strength is relatively well preserved in young patients with cystic fibrosis. However, the strength of the diaphragm decreases with the progression of malnutrition and airway obstruction. PMID- 15531667 TI - Interaction of fat availability and sex on postprandial satiety and cholecystokinin after mixed-food meals. AB - BACKGROUND: Cholecystokinin (CCK) is associated with fat-induced satiety. OBJECTIVE: The primary objective of the present study was to determine, in an acute meal setting, whether the availability of dietary fat for alimentary processing, and hence the stimulation of CCK, affects the postmeal satiety response in men and women. DESIGN: In a within-subjects design, subjects (8 men, 7 women) consumed 1 of 3 isoenergetic mixed-food test meals 1 wk apart in random order. The test meals contained 30% of energy from fat, of which more than two thirds was derived from whole almonds, almond oil, or a mix of safflower and corn oils. Visual analogue scales were used to assess indexes of satiety at defined time points up to 6 h after meal consumption. Blood was sampled at corresponding time points for measurement of CCK, glucose, insulin, and triacylglycerol. Subsequent food intake was also assessed. RESULTS: All meals suppressed hunger and induced a pattern of satiety that was sex-specific and corresponded with the CCK response. Women had higher plasma CCK concentrations and experienced greater satiety after the almond oil and control meals (fat as oil) than after the whole almond meal (fat in whole food structure). Men showed no differential response among meals for CCK and satiety. Plasma triacylglycerol differed by time among meals but not by sex, and no significant differences in glucose and insulin were found. CONCLUSIONS: The satiety response to dietary fat provided in oil or whole food form is influenced by sex and is dependent on the availability of fat to stimulate CCK release in women but not in men. PMID- 15531668 TI - Relation between whole-body and regional measures of human skeletal muscle. AB - BACKGROUND: It is unknown whether regional measures of skeletal muscle (SM) in the thigh and abdomen accurately reflect whole-body SM mass. OBJECTIVE: We aimed to determine whether thigh and abdominal SM measures reflect whole-body SM mass and, if so, which region is a stronger marker. DESIGN: Whole-body and regional measures of SM were obtained by magnetic resonance imaging in a sample of 387 white men and women. RESULTS: The regional SM measures, whether obtained by using a single image (midthigh or L4-L5 level) or a series of 7 consecutive images covering 31 cm (thigh or abdomen), were strongly correlated with whole-body SM (P < 0.001). Independent of sex, the thigh SM measures derived from a single image (men: R(2) = 0.77, SEE = 6.5%; women: R(2) = 0.79, SEE = 7.4%) or a series of 7 consecutive images (men: R(2) = 0.84, SEE = 5.4%; women: R(2) = 0.90, SEE = 5.1%) were stronger correlates of whole-body SM with smaller SEE values than were the abdominal SM measures (P < 0.01). However, SM in the abdomen was also a strong marker of whole-body SM, whether determined from a single image at the L4-L5 level (men: R(2) = 0.63, SEE = 8.2%; women: R(2) = 0.58, SEE = 10.4%) or from a series of images across the abdomen (men: R(2) = 0.77, SEE = 6.5%; women: R(2) = 0.70, SEE = 8.7%). CONCLUSION: Although thigh measures of SM are better predictors of whole-body SM, a single image within the abdomen routinely used to estimate abdominal fat may also be a useful marker of whole-body SM. PMID- 15531669 TI - Truncal fat mass as a contributor to inflammation in end-stage renal disease. AB - BACKGROUND: An activated inflammatory response is a common feature of end-stage renal disease (ESRD) and predicts outcome. Adipose tissue is an endocrine organ that may contribute to an inflammatory burden by secreting adipocytokines such as interleukin 6 (IL-6). OBJECTIVE: The objective was to relate plasma concentrations of IL-6 in ESRD patients to body composition, regional fat mass distribution, and blood lipid profiles. DESIGN: One hundred ninety-seven ESRD patients (123 men; +/- SE age: 52 +/- 1 y) were evaluated shortly before dialysis started. Lean body mass and truncal and nontruncal fat mass were estimated by dual-energy X-ray absorptiometry. Nutritional status was evaluated on the basis of subjective global assessment and handgrip strength. Inflammatory biomarker and blood lipid concentrations were also evaluated. RESULTS: Median IL-6 (8.5 compared with 4.5 pg/mL; P < 0.001) concentrations were significantly greater in malnourished than in well-nourished patients. Moreover, negative correlations were observed between IL-6 and serum creatinine (rho = -0.19, P < 0.01), handgrip strength (rho = -0.24, P < 0.001), and serum albumin (rho = -0.34, P < 0.001). A significantly higher truncal fat mass (12.8 +/- 0.7 compared with 10.5 +/- 0.4 kg; P < 0.005) was observed in ESRD patients with inflammation (C-reactive protein >/= 10 mg/L). Inverse correlations were observed between plasma IL-6 and HDL cholesterol (rho = -0.16, P < 0.05) and apolipoprotein A (rho = -0.23, P < 0.001). CONCLUSIONS: Plausible relations exist between inflammatory biomarkers, such as IL-6 and high-sensitivity C-reactive protein, and regional fat distribution in ESRD patients. Moreover, the strong inverse relations between HDL cholesterol and apolipoprotein A and biomarkers of inflammation suggest that the chronic inflammatory response observed in ESRD patients is an important contributor to the atherogenic lipoprotein profile in uremia. PMID- 15531670 TI - Effects of exercise intensity on food intake and appetite in women. AB - BACKGROUND: Increasing exercise intensity has been shown to reduce energy intake in men. OBJECTIVE: The main objective of this study was to investigate the effects of exercise intensity on energy intake in women. DESIGN: Thirteen moderately active (peak oxygen uptake: 44.0 +/- 4.7 mL . kg(-1) . min(-1)) women [body mass index (in kg/m(2)): 22.2 +/- 2.4; age: 22.2 +/- 2.0 y] were subjected to 3 experimental conditions: control with no exercise and 2 equicaloric (350 kcal) low- (LIE) and high- (HIE) intensity exercise sessions at 40% and 70% of peak oxygen uptake, respectively. After each session, the participants ate ad libitum from buffet-type meals at lunch and dinner and ate snacks during the afternoon and evening. Visual analogue scales were used to rate appetite. RESULTS: More energy was ingested at lunchtime after the HIE session than after the control session (878 +/- 309 and 751 +/- 230 kcal, respectively; P = 0.02). Relative energy intake (postexercise energy intake corrected for the energy cost of exercise above the resting level) at lunch was lower after the LIE session than after the control session (530 +/- 233 and 751 +/- 230 kcal, respectively; P < 0.001) and was lower after the HIE session than after the control session (565 +/- 301 and 751 +/- 230 kcal, respectively; P < 0.01). Similarly, daily energy intake tended to increase during the HIE session relative to that during the control session. No treatment effect was found for appetite scores throughout the experiment. CONCLUSION: The results suggest that HIE increases energy intake in women. PMID- 15531671 TI - Changes in whole-grain, bran, and cereal fiber consumption in relation to 8-y weight gain among men. AB - BACKGROUND: Epidemiologic studies that directly examine changes in whole-grain consumption in relation to weight gain are sparse, and characterization of this association has been obscured by methodologic inconsistencies in the assessment of whole grains. OBJECTIVE: We aimed to ascertain the associations between changes in new quantitative estimates of whole-grain intake and 8-y weight gain among US men. DESIGN: The study was conducted in a prospective cohort of 27 082 men aged 40-75 y at baseline in 1986. Data on lifestyle factors were obtained periodically by using self-reported questionnaires, and participants measured and reported their body weight in 1986 and 1994. RESULTS: In multivariate analyses, an increase in whole-grain intake was inversely associated with long-term weight gain (P for trend < 0.0001). A dose-response relation was observed, and for every 40-g/d increment in whole-grain intake from all foods, weight gain was reduced by 0.49 kg. Bran that was added to the diet or obtained from fortified-grain foods further reduced the risk of weight gain (P for trend = 0.01), and, for every 20 g/d increase in intake, weight gain was reduced by 0.36 kg. Changes in cereal and fruit fiber were inversely related to weight gain. No associations were observed between changes in refined-grain or added germ consumption and body weight. CONCLUSIONS: The increased consumption of whole grains was inversely related to weight gain, and the associations persisted after changes in added bran or fiber intakes were accounted for. This suggests that additional components in whole grains may contribute to favorable metabolic alterations that may reduce long term weight gain. PMID- 15531672 TI - Glycemia and insulinemia in healthy subjects after lactose-equivalent meals of milk and other food proteins: the role of plasma amino acids and incretins. AB - BACKGROUND: Milk products deviate from other carbohydrate-containing foods in that they produce high insulin responses, despite their low GI. The insulinotropic mechanism of milk has not been elucidated. OBJECTIVE: The objective was to evaluate the effect of common dietary sources of animal or vegetable proteins on concentrations of postprandial blood glucose, insulin, amino acids, and incretin hormones [glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1] in healthy subjects. DESIGN: Twelve healthy volunteers were served test meals consisting of reconstituted milk, cheese, whey, cod, and wheat gluten with equivalent amounts of lactose. An equicarbohydrate load of white-wheat bread was used as a reference meal. RESULTS: A correlation was found between postprandial insulin responses and early increments in plasma amino acids; the strongest correlations were seen for leucine, valine, lysine, and isoleucine. A correlation was also obtained between responses of insulin and GIP concentrations. Reconstituted milk powder and whey had substantially lower postprandial glucose areas under the curve (AUCs) than did the bread reference ( 62% and -57%, respectively). Whey meal was accompanied by higher AUCs for insulin (90%) and GIP (54%). CONCLUSIONS: It can be concluded that food proteins differ in their capacity to stimulate insulin release, possibly by differently affecting the early release of incretin hormones and insulinotropic amino acids. Milk proteins have insulinotropic properties; the whey fraction contains the predominating insulin secretagogue. PMID- 15531673 TI - L-Rhamnose increases serum propionate after long-term supplementation, but lactulose does not raise serum acetate. AB - BACKGROUND: Acute ingestion of the unabsorbed sugar l-rhamnose in humans raises serum propionate, whereas acute ingestion of lactulose raises serum acetate. It is not known whether short-chain fatty acid concentrations in urine and feces reflect those in blood. OBJECTIVE: The objective was to test the effects of oral l-rhamnose and lactulose for 28 d on acetate and propionate concentrations in serum, urine, and feces. DESIGN: Eleven subjects ingested 25 g l-rhamnose, lactulose, or d-glucose (control) for 28 d in a partially randomized crossover design. One fecal sample, hourly blood samples, and all urine samples were collected over 12 h on the last day of each phase. RESULTS: The increase in serum propionate was greater after l-rhamnose than after lactulose (P < 0.05). The effect of lactulose on serum acetate was not significant, but lactulose raised the acetate:propionate ratio compared with d-glucose or l-rhamnose in serum (P < 0.005) and urine (P < 0.02). Flatulence was significantly greater after lactulose and l-rhamnose than after d-glucose (P < 0.0001), an effect that lasted 4 wk with lactulose but only 1 wk with l-rhamnose. CONCLUSIONS: This study confirmed that l rhamnose ingestion over 28 d continues to selectively raise serum propionate in humans. Although serum acetate did not increase significantly after lactulose, the serum acetate:propionate ratio was significantly different after l-rhamnose and lactulose, which suggests that these substrates could be used to examine the role of colonic acetate and propionate production in the effect of dietary fiber on lipid metabolism. Changes in the ratio of urinary acetate to propionate reflected those in serum. PMID- 15531674 TI - Relation of body mass index and body fatness to energy expenditure: longitudinal changes from preadolescence through adolescence. AB - BACKGROUND: Although it is widely accepted that weight gain results when energy intake exceeds energy expenditure (EE), how reduced EE contributes to the development of obesity remains unclear. OBJECTIVE: We tested the hypothesis that reduced EE in the premenarcheal period in girls constitutes a risk factor for an increase in relative weight [body mass index (BMI) z score] and percentage of body fat (%BF) during adolescence. DESIGN: We measured EE at study entry in 196 premenarcheal nonobese girls. Resting metabolic rate (RMR) was measured by indirect calorimetry. Total energy expenditure (TEE) was measured by the doubly labeled water method. Activity energy expenditure (AEE) was calculated from RMR and TEE. After the baseline study, girls were followed annually until 4 y after menarche (x+/- SD: 7.1 +/- 2.6 y). At each visit, height, weight, and %BF by bioelectrical impedance were measured. Girls also completed annual food-frequency and activity questionnaires. Linear mixed effects modeling was used to evaluate the longitudinal relation between BMI z score and %BF and measures of baseline EE. RESULTS: We found no significant relation in change in %BF with RMR, AEE, or TEE. We observed a small positive relation between BMI z score and AEE and TEE (P < 0.05) but no significant relation with RMR. When we stratified by parental overweight, the findings were unchanged for RMR. TEE and AEE were positively related to BMI z score in girls of overweight parents. CONCLUSIONS: Our findings suggest that EE in the premenarcheal period is not a risk factor for increases in %BF or BMI z score in girls during adolescence. PMID- 15531675 TI - Effects of oral vitamin E and beta-carotene supplementation on ultraviolet radiation-induced oxidative stress in human skin. AB - BACKGROUND: Ultraviolet radiation (UVR) generates reactive oxygen species in skin that can play a role in skin damage, but reports about the photoprotective properties of oral antioxidant supplements are conflicting. OBJECTIVE: We examined the ability of 2 lipid-soluble antioxidants, vitamin E and beta carotene, to reduce markers of oxidative stress and erythema in human skin exposed to UVR. DESIGN: Sixteen healthy subjects took either alpha-tocopherol (n = 8; 400 IU/d) or beta-carotene (n = 8; 15 mg/d) for 8 wk. Biopsy samples before and after supplementation were taken from unexposed skin and skin 6 h after 120 mJ/cm(2) UVR. The effects of supplements on markers of oxidative stress in skin and the minimal erythema dose to UVR were assessed. RESULTS: Supplementary vitamin E was bioavailable, the plasma concentration increased from 14.0 +/- 0.66 (x +/- SEM) to 18.2 +/- 0.64 mug/mL (P < 0.01), and the skin concentration increased from 0.55 +/- 0.09 to 1.6 +/- 0.19 ng/mg protein (P < 0.01). Supplementary beta-carotene increased plasma concentrations from 1 +/- 0.3 to 2.25 +/- 0.3 mug/mL (P < 0.05), but skin concentrations were undetectable. Before vitamin E supplementation, UVR increased the skin malondialdehyde concentration from 0.42 +/- 0.07 to 1.24 +/- 0.16 nmol/mg protein (P < 0.01), whereas oxidized or total glutathione increased from 9.98 +/- 0.4% to 12.0 +/- 1.0% (P < 0.05). Vitamin E supplementation significantly decreased the skin malondialdehyde concentration, but neither vitamin E nor beta-carotene significantly influenced other measures of oxidation in basal or UVR-exposed skin. CONCLUSIONS: Vitamin E or beta-carotene supplementation had no effect on skin sensitivity to UVR. Although vitamin E supplements significantly reduced the skin malondialdehyde concentration, neither supplement affected other measures of UVR-induced oxidative stress in human skin, which suggested no photoprotection of supplementation. PMID- 15531676 TI - Effects of separate delivery of zinc or zinc and vitamin A on hemoglobin response, growth, and diarrhea in young Peruvian children receiving iron therapy for anemia. AB - INTRODUCTION: Anemia is the most prevalent nutritional deficiency in the world. Attempts to improve iron indexes are affected by deficiency of and interaction between other micronutrients. OBJECTIVE: Our goal was to assess whether zinc added to iron treatment alone or with vitamin A improves iron indexes and affects diarrheal episodes. DESIGN: This was a randomized, placebo-controlled, double blind trial conducted in Peru. Anemic children aged 6-35 mo were assigned to 3 treatment groups: ferrous sulfate (FS; n = 104), ferrous sulfate and zinc sulfate (FSZn; n = 109), and ferrous sulfate, zinc sulfate, and vitamin A (FSZnA; n = 110). Vitamin A or its placebo was supplied only once; iron and zinc were provided under supervision >/=1 h apart 6 d/wk for 18 wk. RESULTS: The prevalence of anemia was 42.97%. The increase in hemoglobin in the FS group (19.5 g/L) was significantly less than that in the other 2 groups (24.0 and 23.8 g/L in the FSZn and FSZnA groups, respectively). The increase in serum ferritin in the FS group (24.5 mug/L) was significantly less than that in the other 2 groups (33.0 and 30.8 mug/L in the FSZn and FSZnA groups, respectively). The median duration of diarrhea and the mean number of stools per day was significantly higher in the FS group than in other 2 groups (P < 0.005). CONCLUSION: Adding zinc to iron treatment increases hemoglobin response, improves iron indexes, and has positive effects on diarrhea. No additional effect of vitamin A was found. PMID- 15531677 TI - Triple fortification of salt with microcapsules of iodine, iron, and vitamin A. AB - BACKGROUND: In many developing countries, children are at high risk of goiter, vitamin A deficiency, and iron deficiency anemia. OBJECTIVE: We aimed to develop a stable, efficacious salt fortified with iodine, iron, and vitamin A. DESIGN: A novel spray-cooling technique was used with hydrogenated palm oil to package potassium iodate, micronized ferric pyrophosphate, and retinyl palmitate into microcapsules (mean particle size: 100 mum). We used the microcapsules to create triple-fortified salt (TFS) with 30 mug I, 2 mg Fe, and 60 mug vitamin A/g salt. After storage trials, we compared the efficacy of TFS with that of iodized salt in a 10-mo, randomized, double-blind trial in goitrous schoolchildren (n = 157) who had a high prevalence of vitamin A deficiency and iron deficiency anemia. RESULTS: After storage for 6 mo, losses of iodine and vitamin A from the TFS were approximately 12-15%, and color was stable. In the TFS group, mean hemoglobin increased by 15 g/L at 10 mo (P < 0.01), iron status indexes and body iron stores improved significantly (P < 0.05), and mean serum retinol, retinol-binding protein, and the ratio of retinol-binding protein to prealbumin increased significantly (P < 0.01). At 10 mo, prevalences of vitamin A deficiency and iron deficiency anemia were significantly lower in the TFS group than in the iodized salt group (P < 0.001). CONCLUSION: Newly developed microcapsules containing iodine, iron, and vitamin A are highly stable when added to local African salt. TFS was efficacious in reducing the prevalence of iron, iodine, and vitamin A deficiencies in school-age children. PMID- 15531678 TI - Use of the deuterated-retinol-dilution technique to monitor the vitamin A status of Nicaraguan schoolchildren 1 y after initiation of the Nicaraguan national program of sugar fortification with vitamin A. AB - BACKGROUND: Nicaragua initiated a national program of vitamin A fortification of its domestic sugar supply starting with the 1999-2000 sugarcane harvest. OBJECTIVE: This study was conducted to document any change in the vitamin A status of a cohort of children during the first year of the program. DESIGN: The vitamin A status of 21 Nicaraguan schoolchildren (mean age: 6.7 y; range: 5.3-9.3 y) was assessed in March 2000 and in March 2001. Total-body vitamin A stores and liver vitamin A concentrations were estimated with the deuterated-retinol dilution (DRD) technique at a dose of 5 mg [(2)H(4)]retinyl acetate at baseline and 5 mg [(2)H(8)]retinyl acetate during the repeat test 1 y later. Plasma retinol and carotenoids were measured by HPLC. RESULTS: Median total-body vitamin A stores increased from 0.33 to 0.72 mmol (P = 0.0001), liver vitamin A concentrations from 0.52 to 0.78 mumol/g (P = 0.0003), and plasma retinol concentrations increased from 0.97 to 1.17 mumol/L (P = 0.01). CONCLUSION: The vitamin A status of Nicaraguan schoolchildren improved during the year after the initial distribution of vitamin A-fortified sugar in Nicaragua. PMID- 15531679 TI - Zinc plus beta-carotene supplementation of pregnant women is superior to beta carotene supplementation alone in improving vitamin A status in both mothers and infants. AB - BACKGROUND: Deficiencies of vitamin A, iron, and zinc are prevalent in women and infants in developing countries. Supplementation during pregnancy can benefit mother and infant. OBJECTIVE: We examined whether supplementation during pregnancy with iron and folic acid plus beta-carotene or zinc or both improves the micronutrient status of mothers and infants postpartum. DESIGN: Pregnant women (n = 170) were supplemented daily only during pregnancy with beta-carotene (4.5 mg), zinc (30 mg), or both or placebo plus iron (30 mg) and folic acid (0.4 mg) in a randomized, double-blind, placebo-controlled trial. Micronutrient status was assessed 1 and 6 mo postpartum. RESULTS: Six months postpartum, plasma retinol concentrations were higher in the women who received zinc during pregnancy than in women who did not. Infants born to mothers supplemented with beta-carotene + zinc had higher plasma retinol concentrations, with the frequency of vitamin A deficiency reduced by >30% compared with the other 3 groups. Breast milk beta-carotene concentrations were higher in all women supplemented with beta carotene, but breast-milk retinol concentrations were higher only in women who received beta-carotene + zinc. Zinc concentrations did not differ among groups in mothers and infants. CONCLUSIONS: Zinc supplementation during pregnancy improved the vitamin A status of mothers and infants postpartum, which indicates a specific role of zinc in vitamin A metabolism. Addition of both beta-carotene and zinc to iron supplements during pregnancy could be effective in improving the vitamin A status of mothers and infants. PMID- 15531680 TI - Multivitamin-mineral supplementation is not as efficacious as is iron supplementation in improving hemoglobin concentrations in nonpregnant anemic women living in Mexico. AB - BACKGROUND: Iron supplements improve hemoglobin status and reduce anemia due to iron deficiency. It is not known whether multiple micronutrient (MM) supplements are as efficacious as are iron supplements alone in improving hemoglobin concentrations. OBJECTIVE: We conducted a randomized, double-blind community trial in Mexico to compare the efficacy of MM supplements containing iron with that of iron alone in improving hemoglobin concentrations in nonpregnant women. DESIGN: Nonpregnant women (n = 158) were recruited from a semirural community in Mexico and were randomly assigned to receive iron alone (60 mg; Fe group) or MM supplements (vitamins A, B complex, C, D, E, and K and iron, zinc, and magnesium; MM group) 6 d/wk in their home for 12 wk. Hemoglobin concentrations were measured in capillary blood samples at baseline and follow-up. RESULTS: The treatment groups (MM: n = 75; Fe: n = 77) did not differ significantly at recruitment in age, schooling, literacy, or socioeconomic status. There were no significant differences between groups in compliance (median: 97.5%), baseline hemoglobin concentrations, or prevalence of anemia (20%). Losses to follow-up (4%) and mean (+/-SD) changes in hemoglobin (MM group: 6.7 +/- 10.6 g/L; Fe group: 7.1 +/- 13.6 g/L) were not significantly different between groups. However, the change in hemoglobin in anemic subjects was greater in the Fe group than in the MM group (P < 0.05 for interaction), and there was no significant difference in nonanemic subjects. CONCLUSIONS: MM supplements may not be as efficacious as is iron alone in improving the hemoglobin status of anemic women. PMID- 15531681 TI - Low ratio of S-adenosylmethionine to S-adenosylhomocysteine is associated with vitamin deficiency in Brazilian pregnant women and newborns. AB - BACKGROUND: Pregnant women with low cobalamin concentrations are unable to provide the necessary amount of cobalamin to their fetuses. The effect of low maternal cobalamin concentrations on transmethylation metabolism in pregnant women and their newborns is unknown. OBJECTIVE: We investigated the relation between maternal and neonatal cobalamin concentrations and changes in total homocysteine (tHcy), S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH). DESIGN: Hematologic data and concentrations of cobalamin, red blood cell folate, serum folate, tHcy, methylmalonic acid, SAM, SAH, and other metabolites were measured in 119 serum specimens from pregnant Brazilian women (gestational age: 37-42 wk) and their newborns' placental veins at the time of delivery. RESULTS: The tHcy concentrations were higher in placental vein serum from newborns whose mothers had low cobalamin. Serum SAH concentrations were elevated and serum SAM and methionine concentrations were decreased in pregnant women with lower cobalamin concentrations. SAM:SAH was significantly decreased in both cobalamin-deficient pregnant women and their newborns. CONCLUSIONS: Lower maternal cobalamin concentrations are associated with higher tHcy and lower SAM:SAH in newborns. Because SAM:SAH is closely linked with the activity of numerous enzymatic methylation reactions, these results suggest that methylation could be impaired in cobalamin-deficient pregnant women and their newborns. PMID- 15531682 TI - Bone mass is recovered from lactation to postweaning in adolescent mothers with low calcium intakes. AB - BACKGROUND: Adolescent mothers may be at increased risk of irreversible bone loss during pregnancy and lactation, particularly when calcium intake is low. OBJECTIVE: Longitudinal changes in bone mass from lactation to postweaning were evaluated in 10 adolescent mothers aged 15-18 y who habitually consumed <500 mg Ca/d. DESIGN: Total-body bone mineral content (TBBMC), total-body bone mineral density (TBBMD), and lumbar spine bone mineral density (LSBMD) were measured at lactation (6-24 wk postpartum) and after weaning (12-30 mo postpartum). Serum hormones (intact parathyroid hormone, estradiol, and prolactin), serum calcium, and markers of bone turnover [urinary N-telopeptide cross-linking region of type I collagen (NTx) and plasma activity of bone alkaline phosphatase] were measured at lactation. RESULTS: TBBMC, total calcium content, TBBMD, and LSBMD increased from lactation to postweaning (P < 0.01). TBBMD and LSBMD were, respectively, 3.6% and 9.7% lower than predicted at lactation and 0.3% and 4.8% lower than predicted in the postweaning period. The increase in age-matched TBBMD adequacy was correlated with the time after resumption of menses (r = 0.86, P < 0.01). Calcium accretion from lactation to postweaning correlated negatively with estradiol (r = -0.86) and prolactin (r = -0.69) and positively with intact parathyroid hormone (r = 0.72) and NTx (r = 0.84) measured at lactation (P < 0.05). CONCLUSIONS: It appears that adolescent mothers with habitually low calcium intakes recover from lactation-associated bone loss after weaning. The rate of bone accretion, however, may not be sufficient to attain peak bone mass at maturity. Hormones regulating bone turnover during lactation may influence bone recovery in adolescent mothers. PMID- 15531683 TI - Boron concentrations in milk from mothers of full-term and premature infants. AB - BACKGROUND: Boron is a bioactive element that satisfies several of the criteria for essentiality in humans. OBJECTIVE: The objective was to establish the profile of boron metabolism in human milk. DESIGN: Lactating mothers of premature (PRT; n = 10, <2000 g birth weight, <37 wk gestation) and full-term (FT; n = 10, >2500 g, 39-41 wk gestation) infants living in St John's, Canada, collected milk samples once a week for 12 wk. Samples were analyzed for boron, copper, iron, selenium, and zinc by atomic emission or absorption spectrometry after wet-ash digestion with nitric acid and hydrogen peroxide in polytetrafluoroethylene tubes. RESULTS: A mixed-model analysis of variance indicated that boron concentrations were stable in full-term (30 and 28 mug/L milk; P = 0.5) but not in preterm (37 and 27 mug/L; P = 0.01) milk between weeks 1 and 12, respectively. As expected, there were reductions in the concentrations of copper (FT: 651 to 360 mug/L, P < 0.0001; PRT: 542 to 425, P = 0.05), iron (FT: 355 to 225 mug/L, P = 0.0003; PRT: 406 to 287, P = 0.002), selenium (FT: 26.9 to 18.6 mug/L, P < 0.0001; PRT: 28.7 to 20.4, P < 0.0001), and zinc (FT: 4060 to 1190 mug/L, P < 0.0001; PRT: 5970 to 1270, P < 0.0001) over time. CONCLUSIONS: The stable milk boron concentrations over time suggest that boron may be under homeostatic control. The patterns of change in copper, iron, selenium, and zinc concentrations in milk differ from those of boron. Further research is needed to elucidate the mechanism of milk boron secretion. PMID- 15531684 TI - Interactions between growth and body composition in children treated with high dose chronic glucocorticoids. AB - BACKGROUND: Glucocorticoid therapy retards growth during childhood and is believed to lead to a Cushingoid body habitus. However, despite prolonged, repeated courses of glucocorticoid, children with steroid-sensitive nephrotic syndrome (SSNS) have almost normal adult height. Little information exists on body composition. OBJECTIVE: We sought to assess the effect of glucocorticoids on height and body composition by comparing children with SSNS with concurrent healthy reference children. We hypothesized that chronic glucocorticoid therapy leads to obesity, decreased lean mass, and distorted distributions of fat and lean. DESIGN: We performed a cross-sectional study of 52 subjects with SSNS (4-21 y) and 259 reference subjects. The evaluation included height, weight, and pubertal status. Fat and lean masses were assessed by dual-energy X-ray absorptiometry in all subjects. Lifetime glucocorticoid exposure was recorded for subjects with SSNS. Outcomes were expressed as SD scores (SDS). RESULTS: Forty one percent of subjects with SSNS were obese [body mass index (BMI) > 95th percentile], but regional fat distribution was normal. Mean total lean mass-for height was 0.43 SD (95% CI: 0.15, 0.72) higher and mean appendicular lean mass for-total-lean-mass was lower (-0.39 SD; 95% CI: -0.64, -0.14) in SSNS compared with reference children. The mean height-SDS in SSNS was -0.08 SD (95% CI: -0.37, 0.21) relative to national reference data, but height-SDS was significantly decreased given the degree of obesity. Height-SDS was positively associated with BMI-SDS among subjects with SSNS. CONCLUSION: Glucocorticoid therapy for SSNS is complicated by obesity and relatively low appendicular lean mass. Overall height SDS is normal because of a mitigating effect of elevated BMI on glucocorticoid induced growth retardation. PMID- 15531685 TI - Breastfeeding and risk of inflammatory bowel disease: a systematic review with meta-analysis. AB - BACKGROUND: It has long been believed that breastfeeding provides protection against ulcerative colitis and Crohn disease. Studies designated to test this hypothesis were conducted without reaching conclusive results. OBJECTIVE: The aim of this meta-analysis was to examine the role of breastfeeding in preventing inflammatory bowel disease and to summarize the evidence gathered about this subject. DESIGN: A meta-analysis was performed on 17 relevant articles that were found by using MEDLINE, EMBASE, the Internet, and articles' references. The publications were fully reviewed and divided, on the basis of their quality, into 3 groups. RESULTS: Studies showed heterogeneous results. The pooled odds ratios of all the 17 reviewed studies, calculated according to the random-effects model, were 0.67 (95% CI: 0.52, 0.86) for Crohn disease and 0.77 (0.61, 0.96) for ulcerative colitis. However, only 4 studies for Crohn disease and 4 for ulcerative colitis were eventually included in the highest quality group. In this group, the pooled odds ratio was 0.45 (0.26, 0.79) for Crohn disease and 0.56 (0.38, 0.81) for ulcerative colitis. CONCLUSIONS: The results of this meta analysis support the hypothesis that breastfeeding is associated with lower risks of Crohn disease and ulcerative colitis. However, because only a few studies were graded to be of high quality, we suggest that further research, conducted with good methodology and large sample sizes, should be carried out to strengthen the validity of these observations. PMID- 15531686 TI - Milk and lactose intakes and ovarian cancer risk in the Swedish Mammography Cohort. AB - BACKGROUND: High intakes of dairy products and of the milk sugar lactose have been hypothesized to increase ovarian cancer risk, but prospective data are scarce. OBJECTIVE: We examined the association between intakes of dairy products and lactose and the risk of total epithelial ovarian cancer and its subtypes. DESIGN: This was a prospective population-based cohort study of 61 084 women aged 38-76 y who were enrolled in the Swedish Mammography Cohort. Diet was assessed in 1987-1990 with the use of a self-administered food-frequency questionnaire. During an average follow-up of 13.5 y, 266 women were diagnosed with invasive epithelial ovarian cancer; 125 of those women had serous ovarian cancer. RESULTS: After adjustment for potential confounders, women who consumed >/=4 servings of total dairy products/d had a risk of serous ovarian cancer (rate ratio: 2.0; 95% CI: 1.1, 3.7; P for trend = 0.06) twice that of women who consumed <2 servings/d. No significant association was found for other subtypes of ovarian cancer. Milk was the dairy product with the strongest positive association with serous ovarian cancer (rate ratio comparing consuming >/=2 glasses milk/d with consuming milk never or seldom: 2.0; 95% CI: 1.1, 3.7; P for trend = 0.04). We observed a positive association between lactose intake and serous ovarian cancer risk (P for trend = 0.006). CONCLUSIONS: Our data indicate that high intakes of lactose and dairy products, particularly milk, are associated with an increased risk of serous ovarian cancer but not of other subtypes of ovarian cancer. Future studies should consider ovarian cancer subtypes separately. PMID- 15531688 TI - Alcohol consumption, 29-y total mortality, and quality of life in men in old age. AB - BACKGROUND: The negative effects of excessive alcohol consumption are well known, but moderate alcohol consumption is advocated for health reasons. OBJECTIVE: We compared 29-y total mortality and quality of life in old age by alcohol consumption in midlife. DESIGN: Cardiovascular disease risk factors and alcohol consumption were assessed in 1974 in 1808 men (aged 40-55 y) of high socioeconomic status. At baseline, the men were without signs of chronic diseases. Baseline alcohol consumption was divided as zero (n = 116), moderate (1 349 g/wk; n = 1519), and high (>349 g/wk; n = 173). Quality of life was surveyed in 2000 with the RAND-36 (SF-36) health survey (n = 1216). Mortality was retrieved from registers during the 29-y follow-up. RESULTS: Median alcohol consumption in 1974 and in 2000 was 123 (interquartile range: 56-238) and 84 (28 168) g/wk, respectively, and was significantly correlated. Values of cardiovascular disease risk factors measured in 1974 increased with increasing alcohol consumption. During the 29-y follow-up, 499 men (27.6%) died; mortality was significantly higher among men with the highest alcohol consumption (37.6%) than in abstainers (25.0%) or in men with moderate (26.7%) consumption. Quality of life was not significantly associated with baseline alcohol consumption in responding survivors but was worst in men with high consumption when deaths during follow-up were accounted for. CONCLUSIONS: In this male cohort of high socioeconomic status, only the highest alcohol consumption (>3 drinks/d) affected mortality, and it was associated with worse quality of life in old age. Moderate alcohol consumption in middle age offered no special benefits compared with abstinence over the long term. PMID- 15531687 TI - Calcium intake and colorectal adenoma in a US colorectal cancer early detection program. AB - BACKGROUND: Calcium can reduce the risk of colorectal tumors by binding secondary bile and fatty acids, which leads to antiproliferative effects in the bowel, or by acting directly on the colonic epithelium, which affects differentiation and apoptosis. OBJECTIVE: We investigated calcium intake and risk of colon adenoma to evaluate the association of calcium intake with early stages of colorectal tumor development. DESIGN: We compared the supplemental and dietary calcium intakes of 3696 participants with histologically verified adenoma of the distal colon (ie, descending colon, sigmoid colon, or rectum) with the calcium intakes of 34 817 sigmoidoscopy-negative control participants in the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Calcium intake was assessed at study entry with a 137-item food-frequency questionnaire and additional questions on the amount and duration of calcium supplement use. RESULTS: After adjustment for known risk factors, adenoma risk was lower by 12% for participants in the highest quintile of total calcium intake (>1767 mg/d) than for participants in the lowest quintile (<731 mg/d) (odds ratio: 0.88; 95% CI: 0.76, 1.02; P for trend = 0.04). The protective association between total calcium and colorectal adenoma was largely due to calcium supplement use, with a 27% decrease in adenoma risk for participants taking >1200 mg/d than for nonusers of supplements (odds ratio: 0.73; 95% CI: 0.56, 0.91; P for trend = 0.005). The protective associations of total and supplemental calcium were strongest for colon adenoma (descending and sigmoid colon). CONCLUSION: High calcium intake, particularly from supplements, is associated with a reduced risk of distal colorectal adenoma. PMID- 15531689 TI - Choice of instrument influences relations between food insecurity and obesity in Latino women. AB - BACKGROUND: The results of studies examining food insecurity and obesity in adults are conflicting. Discrepancies could be due to the use of different instruments or to cultural factors that influence response patterns. OBJECTIVE: The goal was to examine the relation of food insecurity to weight status in low income Latino women. DESIGN: A cross-sectional survey was conducted among 559 low income Latino women selected by convenience sampling. The survey included the 18 item US Household Food Security Scale, 2 items related to current and past food insufficiency, demographic information, and measured heights and weights. Data were collected between February and May 2001 in 6 California counties. The main outcomes were frequency of overweight and obesity, defined by a body mass index (in kg/m(2)) of 25-29.9 (overweight) and >30 (obese). Data analysis included analysis of variance, Mantel-Haenszel chi-square test, and logistic regression. RESULTS: The prevalence of food insecurity was 50-60%, and that of obesity was 37.4%. Controlling for years spent in the United States, per capita income, and parity, food insecurity with hunger, measured by the 10-item adult scale of the Food Security Scale, was significantly related to obesity (OR: 1.98; 95% CI: 1.14, 3.53). No interaction between years spent in the United States and current food insecurity was observed. Current food insufficiency, as measured by a single item, was not related to obesity. However, severe past food insufficiency was related to obesity in the US-born population only. CONCLUSION: Food insecurity appears to be related to obesity in Latino women, but choice of instruments might influence the results. PMID- 15531690 TI - World Health Organization equations have shortcomings for predicting resting energy expenditure in persons from a modern, affluent population: generation of a new reference standard from a retrospective analysis of a German database of resting energy expenditure. AB - BACKGROUND: Reference standards for resting energy expenditure (REE) are widely used. Current standards are based on measurements made in the first part of the past century in various races and locations. OBJECTIVE: The aim of the present study was to investigate the application of the World Health Organization (WHO) equations from 1985 in healthy subjects living in a modern, affluent society in Germany and to generate a new formula for predicting REE. DESIGN: The study was a cross-sectional and retrospective analysis of data on REE and body composition obtained from 2528 subjects aged 5-91 y in 7 different centers between 1985 and 2002. RESULTS: Mean REE varied between 5.63 and 8.07 MJ/d in males and between 5.35 and 6.46 MJ/d in females. WHO prediction equations systematically overestimated REE at low REE values but underestimated REE at high REE values. There were significant and independent effects of sex, age, body mass or fat-free mass, and fat mass on REE. Multivariate regression analysis explained up to 75% of the variance in REE. Two prediction formulas including weight, sex, and age or fat-free mass, fat mass, sex, and age, respectively, were generated in a subpopulation and cross-validated in another subpopulation. Significant deviations were still observed for underweight and normal-weight subjects. REE prediction formulas for specific body mass index groups reduced the deviations. The normative data for REE from the Institute of Medicine underestimated our data by 0.3 MJ/d. CONCLUSIONS: REE prediction by WHO formulas systematically over- and underestimates REE. REE prediction from a weight group-specific formula is recommended in underweight subjects. PMID- 15531691 TI - Soy intake and blood cholesterol concentrations: a cross-sectional study of 1033 pre- and postmenopausal women in the Oxford arm of the European Prospective Investigation into Cancer and Nutrition. AB - BACKGROUND: Clinical trials have suggested that the intake of soy protein reduces blood cholesterol. Few studies have explored this relation in subjects who consume soy as part of their regular diet. OBJECTIVE: In this study, we investigated whether blood cholesterol concentrations are related to the intake of soyfoods in a cohort comprising subjects with a wide variation in soy intake. DESIGN: This cross-sectional study included 1033 pre- and postmenopausal women selected from the Oxford arm of the European Prospective Investigation into Cancer and Nutrition. The sample included 361 nonvegetarians, 570 vegetarians, and 102 vegans. Their dietary intake was assessed by using a food-frequency questionnaire. Anthropometric data, medical history, and lifestyle information were obtained with the use of a questionnaire, blood samples were obtained, and plasma total, LDL-, and HDL-cholesterol concentrations were measured. RESULTS: Soy-protein intake was inversely associated with total and LDL-cholesterol concentrations and with the ratio of total to HDL cholesterol but not with HDL cholesterol concentrations. Mean plasma LDL-cholesterol concentrations in women with a soy-protein intake >/=6 g/d was 12.4% lower than that in women who consumed <0.5 g/d (P < 0.001). CONCLUSION: Moderate intakes of soyfoods as part of a regular diet are associated with favorable blood cholesterol concentrations. PMID- 15531692 TI - Blood pressure and urinary sodium in men and women: the Norfolk Cohort of the European Prospective Investigation into Cancer (EPIC-Norfolk). AB - BACKGROUND: Abundant evidence indicates that a high sodium intake is causally related to high blood pressure, but debate over recommendations to reduce dietary sodium in the general population continues. A key issue is whether differences in usual sodium intake within the range feasible in free-living populations have clinical or public health relevance. OBJECTIVE: We examined the relation between blood pressure and urinary sodium as a marker of dietary intake. DESIGN: This was a study of 23104 community-living adults aged 45-79 y. RESULTS: Mean systolic and diastolic blood pressure increased as the ratio of urinary sodium to creatinine increased (as estimated from a casual urine sample), with differences of 7.2 mm Hg for systolic blood pressure and 3.0 mm Hg for diastolic blood pressure (P < 0.0001) between the top and bottom quintiles. This trend was independent of age, body mass index, urinary potassium:creatinine, and smoking and was consistent by sex and history of hypertension. The prevalence of those with systolic blood pressure >/= 160 mm Hg halved from 12% in the top quintile to 6% in the bottom quintile; the odds ratio for having systolic blood pressure >/= 160 mm Hg was 2.48 (95% CI: 1.90, 3.22) for men and 2.67 (95% CI: 2.08, 3.43) for women in the top compared with the bottom quintile of urinary sodium. Estimated mean sodium intakes in the lowest and highest quintiles were approximately 80 and 220 mmol/d, respectively. CONCLUSIONS: Within the usual range found in a free-living population, differences in urinary sodium, an indicator of dietary sodium intake, are associated with blood pressure differences of clinical and public health relevance. Our findings reinforce recommendations to lower average sodium intakes in the general population. PMID- 15531693 TI - Apolipoprotein E gene promoter -219G->T polymorphism increases LDL-cholesterol concentrations and susceptibility to oxidation in response to a diet rich in saturated fat. AB - BACKGROUND: The apolipoprotein E (APOE) gene promoter polymorphism (-219G-->T) has been associated with increased risk of myocardial infarction, premature coronary artery disease, and decreased plasma apolipoprotein E concentrations. OBJECTIVE: We aimed to determine in healthy subjects whether this polymorphism modifies the susceptibility of LDL to oxidation and the lipid response to the content and quality of dietary fat. DESIGN: Fifty-five healthy men with the APOE3/E3 genotype (7 GG, 38 GT, and 10 TT) completed 3 dietary periods, each lasting 4 wk. The first was a saturated fatty acid (SFA)-rich diet [38% fat-20% SFA and 12% monounsaturated fatty acid (MUFA)-and 47% carbohydrates (CHO)], which was followed by a CHO-rich diet (30% fat-<10% SFA and 12% MUFA-and 55% CHO) or a MUFA-rich diet (38% fat-<10% SFA and 22% MUFA-and 47% CHO) in a randomized crossover design. At the end of each dietary period, LDL oxidation susceptibility, lipids, and lipoproteins were measured. RESULTS: Compared with carriers of the G allele, TT subjects had a significantly (P < 0.05) shorter lag time after the SFA diet. The replacement of the SFA diet by the CHO or MUFA diet induced a greater increase (P < 0.05) in lag time in the TT subjects than in the GG or GT subjects. Carriers of the T allele had higher LDL-cholesterol (P < 0.05) and apolipoprotein B (P < 0.05) plasma concentrations after the SFA diet than did GG subjects. Compared with GG subjects, carriers of the T allele had a significantly (P < 0.05) greater decrease in LDL cholesterol and apolipoprotein B when they changed from the SFA to the CHO diet. CONCLUSION: The -219G-->T polymorphism may partially explain differences in individual responses to diet. PMID- 15531694 TI - Quantitative trait locus determining dietary macronutrient intakes is located on human chromosome 2p22. AB - BACKGROUND: Obesity is generally accompanied by increased food intake. OBJECTIVE: We sought to identify the genes influencing variation in dietary macronutrient intakes in Mexican Americans. DESIGN: We conducted a genome-wide scan by using data derived from food-frequency questionnaires in 816 participants from the San Antonio Family Heart Study. Household effect was simultaneously estimated in a variance component model with the use of SOLAR. RESULTS: All dietary intake measures (total calories, proteins, fat, saturated fat, monounsaturated fat, polyunsaturated fat, carbohydrates, and sucrose) were moderately heritable. Household effect was insignificant except on total calories and sucrose. Suggestive evidence of linkage with saturated fat intake was found on chromosome 2p22 near marker D2S1346 [logarithm of odds (LOD) = 2.62]. Intakes of total calories, fat, protein, and monounsaturated fat were also suggestively linked to the same marker. A significant linkage signal on chromosome 2p22 for leptin concentrations and fat mass was localized in this population, so we used leptin or fat mass as a covariate. Multipoint LOD scores for saturated fat dropped to 1.27 and 1.90, respectively, which suggested that this region on chromosome 2p contributes to both saturated fat intake and body adiposity. This chromosomal region contains the proopiomelanocortin gene (POMC). However, 2 polymorphisms in exon 3 of the POMC gene showed no association with saturated fat intake. CONCLUSIONS: The results strengthen the hypothesis that chromosome 2p22 harbors genes that influence a variety of obesity-related phenotypes, including macronutrient intakes. PMID- 15531695 TI - Calcium absorption in Nigerian children with rickets. AB - BACKGROUND: Nutritional rickets is common in Nigerian children and responds to calcium supplementation. Low dietary calcium intakes are also common in Nigerian children with and without rickets. OBJECTIVE: The objective was to assess intestinal calcium absorption in Nigerian children with rickets. DESIGN: Calcium absorption was assessed in 15 children with active rickets (2-8 y of age) and in 15 age- and sex-matched children without rickets by using a dual-tracer stable isotope method. The children with rickets were supplemented with calcium for 6 mo; calcium absorption was reevaluated 12 mo after the baseline study. Fractional calcium absorption could be determined in 10 children with rickets and in 10 children without rickets. RESULTS: The children with and without rickets had dietary calcium intakes of approximately 200 mg/d. Compared with the control children, the children with rickets had lower serum 25-hydroxyvitamin D and calcium concentrations and greater 1,25-dihydroxyvitamin D and parathyroid hormone concentrations. In fact, there were 15 rachitic and 15 control children in the study. Mean (+/-SD) fractional calcium absorption did not differ between those with (61 +/- 20%) and without (63 +/- 13%) rickets (P = 0.47). Calcium absorption was not associated with serum concentrations of calcium, alkaline phosphatase, 25-hydroxyvitamin D, 1,25-dihydroxyvitamin D, or parathyroid hormone. Mean fractional calcium absorption was significantly greater after (81 +/- 10%) than before (61 +/- 20%) calcium supplementation for the treatment of rickets (P = 0.035). CONCLUSIONS: In Nigerian children with rickets, the capacity to absorb calcium is not impaired; however, fractional calcium absorption increases after the resolution of active disease. Calcium absorption may be inadequate to meet the skeletal demands of children with rickets during the active phase of the disease, despite being similar to that of control children. PMID- 15531696 TI - Effect of habitual dietary calcium intake on calcium supplementation in 12-14-y old girls. AB - BACKGROUND: There is no agreement on how much calcium young girls need for optimal bone mineralization. OBJECTIVE: We evaluated whether the effect of calcium supplementation on whole-body bone mineral accretion depends on habitual calcium intake. DESIGN: This was a randomized, double-blind, placebo-controlled, 1-y calcium intervention study of girls aged 12-14 y selected from a larger group according to habitual calcium intake: subgroup A (n = 60) habitually consumed 1000-1307 mg/d (40th-60th percentile), and subgroup B (n = 53) habitually consumed <713 mg/d (<20th percentile). The girls from each subgroup were randomly assigned to receive either 500 mg Ca/d or placebo. Whole-body bone mineral content (BMC), bone area (BA), bone mineral density (BMD), and BMC adjusted for BA, height, and weight (size-adjusted BMC) were measured at baseline and after 1 y. RESULTS: There was no significant effect modification of baseline habitual calcium intake on the relation between calcium supplementation and height, weight, BMC, size-adjusted BMC, BA, BMD, or alkaline phosphatase. Calcium supplementation had an effect on BMD (0.8%; P = 0.049) and tended to show signs of an effect on size-adjusted BMC (0.5%; P = 0.08). CONCLUSION: A modest effect of calcium supplementation on BMD was shown. However, the effect was independent of habitual calcium intake. PMID- 15531697 TI - Sex differences in the effect of body-composition variables on bone mass in healthy children and adolescents. AB - BACKGROUND: Prophylactic interventions against osteoporosis require a determination of the factors that influence the accumulation of bone mass during growth. OBJECTIVE: The objective was to determine the independent sex-specific contribution of lean mass and fat mass to bone mineral content (BMC), after adjustment for anthropometric variables and lifestyle factors, in healthy children and adolescents. DESIGN: Healthy schoolchildren (184 boys and 179 girls) aged 10-17 y (x+/- SD: 13.0 +/- 2.1 y) participated in this cross-sectional study. Total and regional (lumbar spine, femoral neck, and distal one-third of the radius) BMC and body composition were measured by dual-energy X-ray absorptiometry. RESULTS: A significant effect of anthropometric variables and lifestyle factors on BMC was observed at all skeletal sites. Lean mass and fat mass showed robust correlations with BMC, even after adjustment for anthropometric variables and lifestyle factors. Lean mass contributed to 6-12% of the variance in BMC in boys and to 4-10% in girls. Fat mass accounted for 0.1-2% of BMC variance in boys and to 0.1-6.5% in girls. CONCLUSIONS: Both lean mass and fat mass are consistent predictors of BMC at multiple skeletal sites in healthy children and adolescents. The contribution of lean mass to BMC variance was larger in boys than in girls. In both sexes, the highest contribution of lean mass to BMC was observed at the femoral neck. PMID- 15531698 TI - Determination of iron absorption from intrinsically labeled microencapsulated ferrous fumarate (sprinkles) in infants with different iron and hematologic status by using a dual-stable-isotope method. AB - BACKGROUND: The use of microencapsulated ferrous fumarate sprinkles is a new approach for home fortification. Iron and hematologic status may affect the absorption of iron from sprinkles. OBJECTIVE: The objective was to measure the absorption (corrected erythrocyte incorporation of (57)Fe) of 2 different doses of iron from sprinkles added to a maize-based complementary food provided to infants with different iron and hematologic status. DESIGN: Infants aged 6-18 mo were randomly assigned to receive either 30 (n = 45) or 45 (n = 45) mg elemental Fe as (57)Fe-labeled sprinkles added to a maize-based porridge on 3 consecutive days. A (58)Fe tracer (0.2 mg as ferrous citrate) was also infused intravenously (n = 46). Blood was drawn at baseline and 14 d later to determine erythrocyte incorporation of (57)Fe and (58)Fe by using inductively coupled plasma mass spectrometry. On the basis of hemoglobin and soluble transferrin receptor concentrations, subjects were classified as having iron deficiency anemia (IDA), iron deficiency (ID), or sufficient iron status. RESULTS: There was no significant effect of dose on iron absorption (P > 0.05). Geometric mean iron absorption was 8.25% (range: 2.9-17.8%) in infants with IDA (n = 32), 4.48% (range: 1.1-10.6%) in infants with ID (n = 20), and 4.65% (range: 1.5-12.3%) in iron-sufficient infants (n = 20). Geometric mean iron absorption was significantly higher in infants with IDA than in infants with ID or iron sufficient infants (P = 0.0004); however, there were no significant differences between infants with ID and iron-sufficient infants. CONCLUSION: During infancy, iron absorption from sprinkles in a maize-based porridge meets and surpasses requirements for absorbed iron and is up-regulated in infants with IDA. PMID- 15531699 TI - Whatever happened to the second law of thermodynamics? PMID- 15531700 TI - Is a calorie a calorie? Biologically speaking, no. PMID- 15531701 TI - Fructose misuse, the obesity epidemic, the special problems of the child, and a call to action. PMID- 15531702 TI - Vitamin B-6 status and coronary artery disease. PMID- 15531703 TI - A modified regression model to adjust for intraindividual variation in serum biomarker concentrations. PMID- 15531704 TI - How to consider low serum vitamin D as a risk factor for insulin resistance? PMID- 15531705 TI - Vitamin D, parathyroid hormone, and insulin sensitivity. PMID- 15531706 TI - Edentulism and malnutrition in elderly patients. PMID- 15531707 TI - Characterization of plastidial thioredoxins from Arabidopsis belonging to the new y-type. AB - The plant plastidial thioredoxins (Trx) are involved in the light-dependent regulation of many enzymatic activities, owing to their thiol-disulfide interchange activity. Three different types of plastidial Trx have been identified and characterized so far: the m-, f-, and x-types. Recently, a new putative plastidial type, the y-type, was found. In this work the two isoforms of Trx y encoded by the nuclear genome of Arabidopsis (Arabidopsis thaliana) were characterized. The plastidial targeting of Trx y has been established by the expression of a TrxGFP fusion protein. Then both isoforms were produced as recombinant proteins in their putative mature forms and purified to characterize them by a biochemical approach. Their ability to activate two plastidial light regulated enzymes, NADP-malate dehydrogenase (NADP-MDH) and fructose-1,6 bisphosphatase, was tested. Both Trx y were poor activators of fructose-1,6 bisphosphatase and NADP-MDH; however, a detailed study of the activation of NADP MDH using site-directed mutants of its regulatory cysteines suggested that Trx y was able to reduce the less negative regulatory disulfide but not the more negative regulatory disulfide. This property probably results from the fact that Trx y has a less negative redox midpoint potential (-337 mV at pH 7.9) than thioredoxins f and m. The y-type Trxs were also the best substrate for the plastidial peroxiredoxin Q. Gene expression analysis showed that Trx y2 was mainly expressed in leaves and induced by light, whereas Trx y1 was mainly expressed in nonphotosynthetic organs, especially in seeds at a stage of major accumulation of storage lipids. PMID- 15531708 TI - Analysis of sequence, map position, and gene expression reveals conserved essential genes for iron uptake in Arabidopsis and tomato. AB - Arabidopsis (Arabidopsis thaliana) and tomato (Lycopersicon esculentum) show similar physiological responses to iron deficiency, suggesting that homologous genes are involved. Essential gene functions are generally considered to be carried out by orthologs that have remained conserved in sequence and map position in evolutionarily related species. This assumption has not yet been proven for plant genomes that underwent large genome rearrangements. We addressed this question in an attempt to deduce functional gene pairs for iron reduction, iron transport, and iron regulation between Arabidopsis and tomato. Iron uptake processes are essential for plant growth. We investigated iron uptake gene pairs from tomato and Arabidopsis, namely sequence, conserved gene content of the regions containing iron uptake homologs based on conserved orthologous set marker analysis, gene expression patterns, and, in two cases, genetic data. Compared to tomato, the Arabidopsis genome revealed more and larger gene families coding for the iron uptake functions. The number of possible homologous pairs was reduced if functional expression data were taken into account in addition to sequence and map position. We predict novel homologous as well as partially redundant functions of ferric reductase-like and iron-regulated transporter-like genes in Arabidopsis and tomato. Arabidopsis nicotianamine synthase genes encode a partially redundant family. In this study, Arabidopsis gene redundancy generally reflected the presumed genome duplication structure. In some cases, statistical analysis of conserved gene regions between tomato and Arabidopsis suggested a common evolutionary origin. Although involvement of conserved genes in iron uptake was found, these essential genes seem to be of paralogous rather than orthologous origin in tomato and Arabidopsis. PMID- 15531709 TI - Root-to-shoot transport of sulfate in Arabidopsis. Evidence for the role of SULTR3;5 as a component of low-affinity sulfate transport system in the root vasculature. AB - Xylem transport of sulfate regulates distribution of sulfur in vascular plants. Here, we describe SULTR3;5 as an essential component of the sulfate transport system that facilitates the root-to-shoot transport of sulfate in the vasculature. In Arabidopsis (Arabidopsis thaliana), SULTR3;5 was colocalized with the SULTR2;1 low-affinity sulfate transporter in xylem parenchyma and pericycle cells in roots. In a yeast (Saccharomyces cerevisiae) expression system, sulfate uptake was hardly detectable with SULTR3;5 expression alone; however, cells coexpressing both SULTR3;5 and SULTR2;1 showed substantial uptake activity that was considerably higher than with SULTR2;1 expression alone. The V(max) value of sulfate uptake activity with SULTR3;5-SULTR2;1 coexpression was approximately 3 times higher than with SULTR2;1 alone. In Arabidopsis, the root-to-shoot transport of sulfate was restricted in the sultr3;5 mutants, under conditions of high SULTR2;1 expression in the roots after sulfur limitation. These results suggested that SULTR3;5 is constitutively expressed in the root vasculature, but its function to reinforce the capacity of the SULTR2;1 low-affinity transporter is only essential when SULTR2;1 mRNA is induced by sulfur limitation. Consequently, coexpression of SULTR3;5 and SULTR2;1 provides maximum capacity of sulfate transport activity, which facilitates retrieval of apoplastic sulfate to the xylem parenchyma cells in the vasculature of Arabidopsis roots and may contribute to the root-to-shoot transport of sulfate. PMID- 15531710 TI - Amino acid metabolism in maize earshoots. Implications for assimilate preconditioning and nitrogen signaling. AB - Nitrogen (N) is an essential requirement for kernel growth in maize (Zea mays); however, little is known about how N assimilates are metabolized in young earshoots during seed development. The objective of this study was to assess amino acid metabolism in cob and spikelet tissues during the critical 2 weeks following silking. Two maize hybrids were grown in the field for 2 years at two levels of supplemental N fertilizer (0 and 168 kg N/ha). The effects of the reproductive sink on cob N metabolism were examined by comparing pollinated to unpollinated earshoots. Earshoots were sampled at 2, 8, 14, and 18 d after silking; dissected into cob, spikelet, and/or pedicel and kernel fractions; then analyzed for amino acid profiles and key enzyme activities associated with amino acid metabolism. Major amino acids in the cob were glutamine (Gln), aspartic acid (Asp), asparagine (Asn), glutamate, and alanine. Gln concentrations dropped dramatically from 2 to 14 d after silking in both pollinated and unpollinated cobs, whereas all other measured amino acids accumulated over time in unpollinated spikelets and cobs, especially Asn. N supply had a variable effect on individual amino acid levels in young cobs and spikelets, with Asn being the most notably enhanced. We found that the cob performs significant enzymatic interconversions among Gln, alanine, Asp, and Asn during early reproductive development, which may precondition the N assimilate supply for sustained kernel growth. The measured amino acid profiles and enzymatic activities suggest that the Asn to Gln ratio in cobs may be part of a signal transduction pathway involving aspartate aminotransferase, Gln synthetase, and Asn synthetase to indicate plant N status for kernel development. PMID- 15531711 TI - Can Ca2+ fluxes to the root xylem be sustained by Ca2+-ATPases in exodermal and endodermal plasma membranes? AB - The pathway of Ca2+ movement from the soil solution into the root stele has been a subject of controversy. If transport through the endodermis is assumed to be through the cytoplasm, the limiting factor is believed to be the active pumping of Ca2+ from the cytoplasm into the stele apoplast through the plasma membrane lying on the stele side of the Casparian band. By analogy, for similar transport through the exodermis, the limiting step would be the active pumping into the apoplast on the central cortical side of the layer. Such effluxes are mediated by Ca2+-ATPases. To assess whether or not known Ca2+ fluxes to the stele in onion (Allium cepa) roots could be supported by Ca2+-ATPases, the percentages of total membrane protein particles required to effect the transport were calculated using measured values of membrane surface areas, an animal literature value for Ca2+ ATPase V(max), plant literature values for Ca2+-ATPase K(m), and protein densities of relevant membranes. Effects of a putative symplastic movement of Ca2+ from the exo- or endodermis into the next cell layer, which would increase the surface areas available for pumping, were also considered. Depending on the assumptions applied, densities of Ca2+ pumps, calculated as a percentage of total membrane protein particles, varied tremendously between three and 1,600 for the endodermis, and between 0.94 and 1,900 for the exodermis. On the basis of the data, the possibility of Ca2+ transport through the cytoplasm and membranes of the exodermis and endodermis cannot be discounted. Thus, it is premature to assign an entirely apoplastic pathway for Ca2+ movement from the soil solution to the tracheary elements of the xylem. To verify any conclusion with certainty, more detailed data are required for the characteristics of exo- and endodermal Ca2+-ATPases. PMID- 15531713 TI - How thyroid tumors start and why it matters: kinase mutants as targets for solid cancer pharmacotherapy. AB - Treatment of patients with thyroid cancer is usually successful, and most patients are cured of the disease. However, we do not have effective therapies for patients with invasive or metastatic thyroid cancer if the disease is not surgically resectable and does not concentrate radio-iodine. Conventional external beam radiotherapy and chemotherapy are of marginal benefit. In other types of cancer, new therapies are being developed that take advantage of our knowledge of cancer pathogenesis to interfere with the activity of specific oncoproteins believed to be important in disease causation. Because these approaches are being considered for thyroid cancer, I will briefly describe in this review examples of recent breakthroughs in medical therapy of certain hematological malignancies and some solid tumors using drugs that work in this fashion, focusing in particular on compounds that block the enzymatic activity of specific tyrosine kinase oncoproteins. It should be noted, however, that cancers commonly harbor mutations or other disruptions of many genes, each of which could conceivably play a role in disease pathogenesis. This makes the choice of molecular target a difficult and critical decision if these approaches are to succeed. Here I will argue that priority should be given to blocking the function of oncoproteins activated early in tumor development. We have a fairly good understanding of the genetic changes involved in thyroid cancer initiation, and hence these cancers may prove to be particularly well suited for oncoprotein specific therapies. PMID- 15531712 TI - Molecular bases for sensitivity to tubulin-binding herbicides in green foxtail. AB - We investigated the molecular bases for resistance to several classes of herbicides that bind tubulins in green foxtail (Setaria viridis L. Beauv.). We identified two alpha- and two beta-tubulin genes in green foxtail. Sequence comparison between resistant and sensitive plants revealed two mutations, a leucine-to-phenylalanine change at position 136 and a threonine-to-isoleucine change at position 239, in the gene encoding alpha2-tubulin. Association of mutation at position 239 with herbicide resistance was demonstrated using near isogenic lines derived from interspecific pairings between green foxtail and foxtail millet (Setaria italica L. Beauv.), and herbicide sensitivity bioassays combined with allele-specific PCR-mediated genotyping. Association of mutation at position 136 with herbicide resistance was demonstrated using herbicide sensitivity bioassays combined with allele-specific PCR-mediated genotyping. Both mutations were associated with recessive cross resistance to dinitroanilines and benzoic acids, no change in sensitivity to benzamides, and hypersensitivity to carbamates. Using three-dimensional modeling, we found that the two mutations are adjacent and located into a region involved in tubulin dimer-dimer contact. Comparison of three-dimensional alpha-tubulin models for organisms with contrasted sensitivity to tubulin-binding herbicides enabled us to propose that residue 253 and the vicinity of the side chain of residue 251 are critical determinants for the differences in herbicide sensitivity observed between organisms, and that positions 16, 24, 136, 239, 252, and 268 are involved in modulating sensitivity to these herbicides. PMID- 15531714 TI - Screening of six risk exons of the RET proto-oncogene in families with medullary thyroid carcinoma in the Czech Republic. AB - Medullary thyroid carcinoma (MTC) occurs as a sporadic form (75%) or as an autosomal dominant inherited familial disorder (25%) called familial MTC (FMTC) or as multiple endocrine neoplasia type 2 (MEN2) syndromes. Germ-line mutations in the rearranged during transfection (RET) proto-oncogene in exons 10, 11, 13, 14, 15 and 16 are known to be a cause of most of the familial forms. In this paper we report molecular genetic testing of 106 families with MTC (358 tested persons) from the Czech Republic in which we directly sequenced these six exons of the RET proto-oncogene. We detected germ-line mutations in 100% of MEN2B families (4/4 families), 90% of MEN2A families (9/10), 40% of FMTC families (4/10) and 7% of apparently sporadic MTC (6/82). Eleven different germ-line mutations were revealed. MEN2B was associated with mutation Met918 Thr in exon 16. In one MEN2B family beside this mutation the Tyr791 Phe was also found, which has not yet been reported. MEN2A was restricted to different mutations in exon 11 (codon 634). In FMTC and 'sporadic' MTC families the mutations in exons 10, 11, 13 and 14 were detected. The genotype/phenotype correlations are given. Genetic testing revealed germ-line mutations in 23 index patients, 24 family members and excluded them in 53 relatives. PMID- 15531715 TI - Has the mammary gland a protective mechanism against overexposure to triiodothyronine during the peripartum period? The prolactin pulse down-regulates mammary type I deiodinase responsiveness to norepinephrine. AB - Peripartum is a crucial period for mammary gland final differentiation and the onset of lactation. Although the 'trigger' for lactogenesis depends on several hormones, a key factor is the peripartum prolactin (PRL) pulse whose deletion results in a failure to initiate milk production. Other hormones having a critical role during this period but exerting a contrary effect are the thyronines. A transitory hypothyroidism occurs at peripartum in serum and several other extrathyroidal tissues, whereas the induction of hyperthyroidism during late pregnancy is associated with the absence of lactation after delivery. We analyzed the mammary gland during pregnancy and lactation for: (a) the type and amount of thyroid receptors (TRs), (b) the local triiodothyronine (T3) generation catalyzed by type I deiodinase (Dio1), (c) the Dio1 response to norepinephrine (NE) and (d) the effect on Dio1 and TRs of blocking the PRL pulse at peripartum. Our data showed that during pregnancy the mammary gland contains Dio1 in low amounts associated with the highest expression of TRalpha1; whereas during lactation the gland shows high levels of both Dio1 and TRalpha1. However, at peripartum, both TRs and Dio1 decrease, and Dio1 becomes refractory to NE. This refractoriness disappears when the PRL pulse is blocked by the dopamine agonist bromocriptine. This blockade is also accompanied by a significant decrease in cyclin D1 expression. Our data suggested that the peripartum PRL pulse is part of a protective mechanism against precocious differentiation and/or premature involution of the alveolar epithelium due to T3 overexposure. PMID- 15531716 TI - Ob receptor in rabbit ovary and leptin in vitro regulation of corpora lutea. AB - We studied leptin involvement in rabbit corpora lutea (CL) activity, and its post transcriptional signalling pathway. The expression of leptin receptor (Ob-R) in rabbit ovary at day 9 of pseudopregnancy was evaluated by immunohistochemistry and Western blot analysis. The specificity of the Ob-R receptor antibodies was characterised by immunoprecipitation and competition with blocking peptide. Day 9 CL were incubated in vitro with leptin alone or with inhibitors of PLC (phospholipase C), PLD (phospholipase D), AC (adenylate cyclase), JAK (janus kinase), MAPK (mitogen-activated protein kinase) and both cAMP- and cGMP-specific PDE (phosphodiesterase). Prostaglandin F2alpha(PGF2alpha), PGE2 and progesterone levels were measured in the culture medium, while NOS (nitric oxide synthase) and cAMP- and cGMP- specific PDE activities were measured in CL tissue. Positive staining for Ob-R was found within the cytoplasm of large luteal cells of CL as well as in granulosa cells of follicles and oocytes. Immunoblots detected a band of about 99 kDa size in Ob-R immunoprecipitates from CL homogenates. This band was not detectable after pre-incubation of the primary antibody with the immunising leptin peptide. Leptin increased PGF2alphaand cAMP-specific PDE, decreased basal progesterone and did not affect PGE2 and NOS levels. Leptin used the JAK pathway in increasing PGF2alpha, and MAPK and cAMP-specific PDE in decreasing progesterone. This study supports a permissive luteolytic role for leptin in rabbit CL. PMID- 15531717 TI - Characterization of the resistance to the anorectic and endocrine effects of leptin in obesity-prone and obesity-resistant rats fed a high-fat diet. AB - Leptin produced by adipocytes controls body weight by restraining food intake and enhancing energy expenditure at the hypothalamic level. The diet-induced increase in fat mass is associated with the presence of elevated circulating leptin levels, suggesting the development of resistance to its anorectic effect. Rats, like humans, show different susceptibility to diet-induced obesity. The aim of the present study was to compare the degree of leptin resistance in obesity-prone (OP) vs obesity-resistant (OR) rats on a moderate high-fat (HF) diet and to establish if the effects of leptin on hypothalamo-pituitary endocrine functions were preserved. Starting from 6 weeks after birth, male Sprague-Dawley rats were fed on either a commercial HF diet (fat content: 20% of total calorie intake) or a standard pellet chow (CONT diet, fat content: 3%). After 12 weeks of diet, rats fed on HF diet were significantly heavier than rats fed on CONT diet. Animals fed on HF diet were ranked according to body weight; the two tails of the distribution were called OP and OR rats respectively. A polyethylene cannula was implanted into the right ventricle of rats 1 week before central leptin administration. After 12 weeks of HF feeding, both OR and OP rats were resistant to central leptin administration (10 mug, i.c.v.) (24 h calorie intake as a percent of vehicle-treated rats: CONT rats, 62 [50; 78]; OR, 93 [66; 118]; OP, 90 [70; 120] as medians and 95% confidence intervals (CIs) of six rats for each group). Conversely, after 32 weeks of diet both OR and OP rats were partially responsive to 10 mug leptin i.c.v. as compared with CONT rats (24 h calorie intake as a percent of vehicle-treated rats: CONT rats, 60 [50; 67]; OR, 65 [50; 80]; OP, 80 [60; 98] as medians and 95% CIs of six rats for each group); the decrease of food intake following 200 mug leptin i.p. administration was similar in all the three groups (calorie intake as a percent of vehicle-treated rats: 86 [80; 92] as median and 95% CI). The long-term intake of HF diet caused hyperleptinemia, hyperinsulinemia and higher plasma glucose levels in OP rats as compared with CONT rats. Plasma thyroxine (T4) was lower in all the rats fed the HF diet as compared with CONT. i.c.v. administration of leptin after 32 weeks of diet restored normal insulin levels in OP rats. Moreover, leptin increased plasma T4 concentration and strongly enhanced GH mRNA expression in the pituitary of OP as well as OR rats (180+/-10% vs vehicle-treated rats). In conclusion, long-term intake of HF diet induced a partial central resistance to the anorectic effect of leptin in both lean and fat animals; the neuroendocrine effects of leptin on T4 and GH were preserved. PMID- 15531718 TI - Metformin inhibits leptin secretion via a mitogen-activated protein kinase signalling pathway in brown adipocytes. AB - Metformin is an anti-diabetic drug with anorexigenic properties. The precise cellular mechanisms of its action are not entirely understood. Adipose tissue has recently been recognized as an important endocrine organ that is pivotal for the regulation of insulin resistance and energy homeostasis. Due to its thermogenic capacity brown adipose tissue contributes to the regulation of energy metabolism and is an attractive target tissue for pharmacological approaches to treating insulin resistance and obesity. Leptin is the prototypic adipocyte-derived hormone inducing a negative energy balance. We investigated effects of metformin on adipocyte metabolism, signalling, and leptin secretion in a brown adipocyte model. Metformin acutely stimulated p44/p42 mitogen-activated protein (MAP) kinase in a dose- (3.2-fold at 1 mmol/l, P< 0.05) as well as time-dependent (3.8 fold at 5 min, P< 0.05) manner. This stimulation was highly selective since phosphorylation of intermediates in the stress kinase, janus kinase (JAK)-signal transducer and activator of transcription (STAT), and phosphatidylinositol (PI) 3 kinase signalling pathways such as p38 MAP kinase, STAT3, and Akt was unaltered. Furthermore, chronic metformin treatment for 12 days dose-dependently inhibited leptin secretion by 35% and 75% at 500 mumol/l and 1 mmol/l metformin respectively (P< 0.01). This reduction was not caused by alterations in adipocyte differentiation. Moreover, the impairment in leptin secretion by metformin was reversible within 48 h after removal of the drug. Pharmacological inhibition of p44/p42 MAP kinase prevented the metformin-induced negative effect on leptin secretion. Taken together, our data demonstrate direct acute effects of metformin on adipocyte signalling and endocrine function with robust inhibition of leptin secretion. They suggest a selective molecular mechanism that may contribute to the anorexigenic effect of this antidiabetic compound. PMID- 15531719 TI - Comparative effects of amino acids and glucose on insulin secretion from isolated rat or mouse islets. AB - Glucose and the combination of leucine and glutamine were used to stimulate insulin secretion from rat islets during a dynamic perifusion and the responses obtained were compared with those elicited from mouse islets under identical conditions. In rat islets, glucose (15 mM) or the amino acid combination of 10 mM glutamine plus 20 mM leucine were most efficacious and peak second-phase insulin release responses were 20- to 30-fold above prestimulatory rates. In contrast to rat islet responses, sustained second-phase insulin secretory responses to the same agonists were minimally increased 1- to 2-fold from mouse islets. Parallel studies demonstrated that phospholipase C (PLC) was markedly activated in rat, but not mouse, islets by both high glucose concentrations and the amino acid combination. Additional studies documented that glucose and amino acid responses of both rat and mouse islets were amplified by carbachol or forskolin. However, wortmannin, a phosphatidylinositol 3-kinase inhibitor, amplified only the responses to glucose leaving the responses to the amino acid mixture unaltered. These observations support the concept that mitochondrial metabolism alone is minimally effective in stimulating insulin secretion from islets. The activation of the supplementary second messenger systems (PLC and/or cAMP) appears essential for the emergence of their full secretory potential. The mechanism regulating the potency and specificity of wortmannin's impact on glucose-induced secretion remains to be identified; however a unique mechanism is supported by these findings. PMID- 15531720 TI - Islet neogenesis: an apparent key component of long-term pancreas adaptation to increased insulin demand. AB - This study aimed to determine the relative importance of different functional and morphological pancreatic changes induced by the chronic administration of a sucrose-rich diet (SRD) to maintain normal glucose homeostasis. Male Wistar rats were fed either sucrose (SRD) or starch (CD) for 6 and 12 months. At both periods, serum glucose and triacylglycerol levels were significantly higher (P<0.05; paired and unpaired Student's t-test) in SRD rats. Serum insulin levels were significantly lower in SRD only at 12 months. At 6 months, the insulin secretion dose-response curve in SRD rats showed a shift to the left that was no longer observed at 12 months, when SRD islets decreased their response to 16 mM glucose. At 6 months, SRD rats showed a significant increase in beta-cell volume density (Vvi) and islet cell replication rate, together with a decrease in beta cell apoptotic rate. Changes were not detected in the percentage of PDX-1- and islet neogenesis associated protein (INGAP)-positive cells. Conversely, at 12 months, there was a significant decrease in beta-cell Vvi and in the percentage of PDX-1-positive cells; the islet cell replication rate was not modified, and the number of apoptotic beta-cells increased significantly. No signs of increased neogenesis or INGAP-positive cells were recorded at any period in SRD rats. Our results show that SRD rats are unable to develop functional and morphological pancreatic reactive changes sufficient to maintain normal glucose and triacylglycerol levels for a long period. Such failure could be ascribed to their inability to increase the rate of neogenesis and of INGAP production. PMID- 15531721 TI - Androgen generation in adipose tissue in women with simple obesity--a site specific role for 17beta-hydroxysteroid dehydrogenase type 5. AB - Women with polycystic ovary syndrome (PCOS) have high circulating androgens, thought to originate from ovaries and adrenals, and frequently suffer from the metabolic syndrome including obesity. However, serum androgens are positively associated with body mass index (BMI) not only in PCOS, but also in simple obesity, suggesting androgen synthesis within adipose tissue. Thus we investigated androgen generation in human adipose tissue, including expression of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) isozymes, important regulators of sex steroid metabolism. Paired omental and subcutaneous fat biopsies were obtained from 27 healthy women undergoing elective abdominal surgery (age range 30-50 years; BMI 19.7-39.2 kg/m(2)). Enzymatic activity assays in preadipocyte proliferation cultures revealed effcient conversion of androstenedione to testosterone in both subcutaneous and omental fat. RT-PCR of whole fat and preadipocytes of subcutaneous and omental origin showed expression of 17beta-HSD types 4 and 5, but no relevant expression of 17beta-HSD types 1, 2, or 3. Microarray analysis confirmed this expression pattern (17beta-HSD5>17beta-HSD4) and suggested a higher expression of 17beta-HSD5 in subcutaneous fat. Accordingly, quantitative real-time RT-PCR showed significantly higher expression of 17beta-HSD5 in subcutaneous compared with omental fat (P<0.05). 17beta-HSD5 expression in subcutaneous, but not omental, whole fat correlated significantly with BMI (r=0.51, P<0.05). In keeping with these findings, 17beta-HSD5 expression in subcutaneous fat biopsies from six women taking part in a weight loss study decreased significantly with weight loss (P<0.05). A role for 17beta-HSD5 in adipocyte differentiation was further supported by the observed increase in 17beta-HSD5 expression upon differentiation of stromal preadipocytes to mature adipocytes (n=5; P<0.005), which again was higher in cells of subcutaneous origin. Functional activity of 17beta-HSD5 also significantly increased with differentiation, revealing a net gain in androgen activation (androstenedione to testosterone) in subcutaneous cultures, contrasting with a net gain in androgen inactivation (testosterone to androstenedione) in omental cultures. Thus, human adipose tissue is capable of active androgen synthesis catalysed by 17beta-HSD5, and increased expression in obesity may contribute to circulating androgen excess. PMID- 15531722 TI - Influence of dietary arginine on the anabolic effects of androgens. AB - Feeding mice an arginine-deficient diet decreased plasma concentrations of arginine, citrulline and ornithine in the females and arginine in the males, abolishing the sexual dimorphic pattern of these amino acids found in mice fed the standard diet. In addition, the restriction of dietary arginine produced a marked decrease in body and renal weights as well as in the activity of renal ornithine decarboxylase, decreases that were gender dependent since they were observed exclusively in males. The fact that these changes were not associated with the decrease in the circulating levels of testosterone and that the dietary arginine restriction prevented the body weight gain induced by testosterone treatment of female mice fed the standard diet indicates that dietary arginine is required for the anabolic action of androgens. Moreover, under certain conditions that could compromise the renal synthesis of arginine, as in the compensatory renal hypertrophy that follows unilateral nephrectomy, the myotrophic effect of testosterone was transiently impaired. The results also revealed that arginine deficiency produced an opposite effect in the expression of IGF-I and IGF-binding protein 1 in the liver and kidney. Taken together, our results indicate that dietary arginine may be relevant to the anabolic action of testosterone, and suggest that this effect may be mediated by changes in the insulin-like growth factor system. PMID- 15531723 TI - QSOX sulfhydryl oxidase in rat adenohypophysis: localization and regulation by estrogens. AB - The expression of the rat quiescin sulfhydryl oxidase (rQSOX) and its putative regulation by estrogens were investigated in the adenohypophysis. Immunohistochemical observations revealed that rQSOX protein is abundantly expressed throughout the anterior lobe of the pituitary, and can be found in almost all the different cell populations. However, as shown by double immunohisto-chemistry, the cells displaying the strongest rQSOX labeling belong to a subset of gonadotrophs. Immunoelectron microscopy showed that, in adenohypophyseal cells, the protein is linked to the membranes of the rough endoplasmic reticulum, the Golgi apparatus and to dense-core secretory granules. These results are consistent with the secretion of the protein and its presumed role in the extracellular matrix. According to its sulfhydryl oxidase function, rQSOX could also participate in the intracellular folding of secreted proteins or hormones like LH and FSH and act as an endogenous redox modulator of hormonal secretion. A semiquantitative RT-PCR analysis of rQSOX level across the estrous cycle and the fact that chronic administration of 17 beta-estradiol to ovariectomized rats led to a sustained up-regulation of rQSOX in the pituitary suggest that rQSOX expression is controlled by sex hormone levels. Further investigations are needed in order to elucidate its precise roles in that gland and the mechanisms of its regulation. PMID- 15531724 TI - Glucocorticoid down-regulation of rat glucocorticoid receptor does not involve differential promoter regulation. AB - The level of expression of the glucocorticoid receptor (GR) is the principal determinant of glucocorticoid sensitivity in most cells. GR levels are permanently 'set' in a tissue-specific manner in response to the perinatal environment, an effect we have previously shown to relate to differential expression of tissue-enriched alternative promoters/exons 1 of the GR gene. In adult animals, GR levels are dynamically regulated around the 'set point' by glucocorticoids themselves, with glucocorticoids down-regulating GR mRNA in most cells and tissues. Here we have examined whether autoregulation of GR mRNA by glucocorticoids involves differential promoter regulation. We show that, in contrast to tissue-specific programming of GR mRNA levels, autoregulation of GR mRNA in vivo does not involve differential regulation of variant exon 1 containing GR mRNAs in that the major variants are down-regulated to a similar extent by glucocorticoid treatment. Consistent with this, transfections of reporter constructs showed that the majority of GR promoters, which are contained within a 4.4 kb region upstream of exon 2, are similarly regulated by glucocorticoids, with two regions of the promoter redundantly required for glucocorticoid regulation. Thus transcriptional autoregulation can occur in adult tissues around the set point established by promoter selection in early life. PMID- 15531725 TI - Analysis of co-factor function in a glucocorticoid-resistant small cell carcinoma cell line. AB - Human small cell lung carcinoma (SCLC) tumours exhibit neuroendocrine differentiation, secreting hormones such as ACTH and related peptides. While glucocorticoids inhibit ACTH secretion from the pituitary, this does not occur in SCLC tumours and SCLC cell lines. Failure of glucocorticoids to suppress ACTH peptides is accompanied by a global lack of glucocorticoid action in a number of SCLC cell lines. In the human SCLC cell line, COR L103, activation of a human tyrosine aminotransferase (TAT3)-luciferase reporter gene is resistant to glucocorticoids despite similar glucocorticoid receptor (GR) expression to the glucocorticoid-sensitive A549 human lung cancer cell line; moreover, the GR is free of deleterious mutations. Over-expression of a wild-type GR restores glucocorticoid regulation of TAT3-luciferase, and this is enhanced when the activation function (AF)-2 domain is deleted but much reduced when the AF-1 domain is deleted. This suggests aberrant AF-2 activation domain function. We identified defective steroid receptor co-activator 1 (SRC1) recruitment to the GR AF-2 in COR L103 cells, although SRC1 was successfully recruited to the steroid X receptor with rifampicin, suggesting a defect in the GR. Analysis of other GR C terminal co-factors identified increased expression of nuclear co-repressor (NCoR) in COR L103 cells. To determine the impact of this, NCoR was over expressed in A549 cells, where it reduced GR transactivation by 55%. In summary, glucocorticoid resistance is associated with altered SRC protein recruitment and increased expression of NCoR in these SCLC cells, suggesting that glucocorticoid sensitivity may be modified by subtle changes in co-factor recruitment. PMID- 15531726 TI - Expression of epidermal growth factor receptor in neoplastic pituitary cells: evidence for a role in corticotropinoma cells. AB - The oncogenic effects of epidermal growth factor (EGF) have long been established. EGF receptor (EGFr) is overexpressed in many types of tumors and constitutes a target for cancer treatment. The pituitary gland is a target of EGF action and it is very likely that EGFr plays a role in pituitary tumor formation and progression. However, there is a controversy in the literature concerning EGFr expression in the different types of pituitary adenomas. In the present study we investigated the expression pattern of the wild type EGFr (EGFrWT) and the constitutively active variant III (EGFrvIII) at the mRNA and protein levels in a large series of pituitary tumors. EGFrWT was found in a high percentage of hormone-secreting tumors, but only in a small fraction of non-functioning pituitary adenomas, while no expression of the EGFrvIII could be detected by nested RT-PCR in any tumor. Among the hormone-secreting adenomas, the highest incidence of EGFr expression was found in Cushing's pituitary adenomas. Furthermore, immunohistochemistry for the phosphorylated EGFr revealed the presence of activated EGFr in most Cushing's adenomas, compared with most pituitary adenomas. Taking into account that downregulation of p27/Kip1 plays a significant role in corticotrope tumorigenesis and that EGFr mitogenic signaling results in decreased p27/Kip1, we searched for a correlation between EGFr expression and p27/Kip1 levels in corticotropinomas. Low p27/Kip1 immunoreactivity was observed in corticotropinomas expressing EGFr. On the other hand, somatotropinomas expressing EGFr had high p27/Kip1 immunoreactivity. These data suggest a corticotrope-specific phenomenon and indicate that EGFr may have a role in the unbalanced growth of corticotrope tumoral cells. PMID- 15531727 TI - All trans-retinoic acid acts synergistically with hydroxytamoxifen and transforming-growth factor beta to stimulate apoptosis in MCF-7 breast cancer cells. AB - The anti-estrogen 4-hydroxytamoxifen (TAM) and vitamin A-related compounds, the retinoids, in combination act synergistically to inhibit growth of breast cancer cells in vitro and in vivo. To clarify the mechanism of this synergism, the effect of TAM and all trans-retinoic acid (AT) on proliferation of MCF-7 breast cancer cells was studied in vitro. TAM and AT acted synergistically to cause a time-dependent and dose-dependent inhibition of MCF-7 cell growth. In a temporally related manner, TAM+AT acted synergistically to downregulate Bcl-2 mRNA and Bcl-2 protein expression, and to stimulate apoptosis. TAM and AT each blocked cell cycle progression throughout 7 days of treatment but without any synergistic or additive effect on this process, indicating a selective synergism for apoptosis. The negative growth factor-transforming growth factor beta (TGFbeta) is secreted by these cells and was studied as a potential mediator of the synergistic effects of TAM+AT on apoptosis. TAM+AT acted synergistically to induce a fivefold increase in TGFbeta1 secretion over 72 h. TGFbeta1 alone had no apoptotic effects on these cells; however, TGFbeta1 in combination with AT acted synergistically to inhibit growth, to downregulate Bcl-2 mRNA and Bcl-2 protein expression, and to stimulate apoptosis of these cells in a manner comparable with that noted for TAM+AT. The synergism of both TAM+AT and TGFbeta1+AT for apoptosis was suppressed by estradiol. Co-incubation of TAM+AT with anti-TGFbeta antibody did not block down-regulation of Bcl-2 protein expression or stimulation of apoptosis. The synergistic effects of TAM+AT on apoptosis therefore occur independently of TGFbeta, although TGFbeta may interact with AT in a novel manner to provide another important anti-proliferative mechanism for breast cancer cells. PMID- 15531728 TI - Gene expression and protein localisation for activin-A, follistatin and activin receptors in goat ovaries. AB - We studied the protein and mRNA expression of activin-A, follistatin and activin receptors in goat ovaries to find evidence of their possible role in ovarian activity, particularly in the various stages of follicle development. Ovaries of cyclic goats were collected and then either fixed in paraformaldehyde for immunohistochemical localisation of activin-A, follistatin, activin receptors IIA/B (ActR-IIA/B) and IA (ActR-IA) proteins or used to obtain samples to demonstrate mRNA expression of activin-A (betaA subunit), follistatin, ActR-IIA, IIB, -IA and -IB, using RT-PCR. For this latter goal, primordial, primary and secondary follicles were isolated mechanically, washed to remove the stromal cells and then used for RT-PCR. In addition, oocytes, cumulus, mural granulosa and theca cells from small (<3 mm) and large (3-6 mm) antral follicles, luteal cells and surface epithelium were collected to study mRNA expression. Activin-A and follistatin proteins were found in oocytes of all follicle classes, granulosa cells from the primary follicle stage onwards, theca cells of antral follicles, corpora lutea and ovarian surface epithelium. In antral follicles, these proteins were detected both in cumulus and mural granulosa cells. ActR-IIA/B protein was found at the same follicular sites, and also in granulosa cells of primordial follicles onward. The localisation of ActR-IA corresponded with that of ActR IIA/B, but the former protein was absent in the theca of large antral follicles. The mRNAs for activin-A (betaA subunit), follistatin, and ActR-IIA, -IIB, -IA and -IB were detected at all follicular and cellular types studied, except that ActR IIB was not found in follicles that had not developed an antrum yet. In conclusion, in goat ovaries, transcripts of activin-A (betaA subunit), its receptors and its binding protein follistatin are expressed and their proteins formed at all follicular stages and in corpora lutea. These findings indicate a role of activin-A in the local regulatory system during the entire follicular development and during luteal activity. PMID- 15531729 TI - Evidence that lactotrophs do not differentiate directly from somatotrophs during chick embryonic development. AB - It is generally accepted that, in mammals, lactotrophs differentiate from somatotrophs through an intermediate cell type, the mammosomatotroph. However, little information exists about mammosomatotrophs and their relationship with lactotroph development in non-mammalian vertebrates. We reported previously that corticosterone (CORT) can induce both somatotroph and lactotroph differentiation in cultures of chicken embryonic pituitary cells. Our current objectives were to determine the abundance of mammosomatotrophs during chicken pituitary development, to identify mammosomatotrophs during CORT induction of lactotrophs, and to explore whether lactotrophs induced by CORT are derived from somatotrophs. Cells that produced prolactin (PRL) only, growth hormone (GH) only or both hormones simultaneously were detected by three approaches - dual immunofluorescence, a combination of immunofluorescence and immunocytochemistry (ICC), and by ICC using combinations of antibodies to GH and PRL. Mammosomatotrophs were not detected between embryonic day (E) 16 and E20, even though lactotrophs increased from nearly absent to greater than 10% of all pituitary cells during this period. CORT induced more than 10% of all E13 pituitary cells to produce PRL, while the percentage of mammosomatotrophs remained at less than 1% of all cells. When cells from the cephalic and caudal lobes of the anterior pituitary were treated separately, CORT increased GH cells in cultures from the caudal lobe. No PRL cells were found in the caudal lobe. In the cephalic lobe, CORT increased lactotrophs, while GH cells were barely detected. In summary, mammosomatotrophs are rare during chicken pituitary development, and CORT does not induce lactotrophs from somatotrophs. These findings indicate that, unlike in mammals, lactotrophs do not differentiate from somatotrophs during chicken embryonic development. PMID- 15531730 TI - Alterations of somatotropic function in prion disease in sheep. AB - This study aimed at investigating the possible linkage between natural scrapie and alterations of the somatotropic axis. Scrapie-affected ewes exhibited 2-fold higher mean GH concentrations during both autumn and spring. GH pulse frequencies were higher in scrapie-affected ewes than in control animals (mean+/-S.E.M. number of pulses/24 h: 10.4+/-0.9 and 7.6+/-0.9 for scrapie-affected and control ewes respectively) suggesting the involvement of central mechanisms. GH secretion induced by administration of an alpha(2)-adrenergic agonist, which acts centrally to stimulate GH secretion, was similar between healthy and scrapie-affected ewes (ratios of the area under the curve (AUC) of GH concentration after to the GH AUC before the agonist administration were 3.6+/-1.6 and 4.9+/-1.0 for scrapie affected and control ewes respectively). Finally, humoral markers and parameters of the metabolic status were determined to test the hypothesis that scrapie associated alterations of GH secretion could be related to disruption of metabolic homeostasis. Glucose, insulin and urea plasma concentrations were higher in scrapie-affected than in healthy ewes. Neither leptin nor IGF-I levels were affected by scrapie. Total thyroxine (T4) was decreased in scrapie-affected ewes but free T4 and total and free triiodothyronine were not modified. In conclusion, our results showed the existence in scrapie-affected ewes of endocrine and metabolic alterations typical of acute illness proceeding, at least in part, from central mechanisms. PMID- 15531731 TI - Cellular topoisomerase I inhibition and antiproliferative activity by MJ-III-65 (NSC 706744), an indenoisoquinoline topoisomerase I poison. AB - To overcome camptothecin's (CPT) lactone instability, reversibility of the drug target interaction, and drug resistance, attempts to synthesize compounds that are CPT-like in their specificity and potency yet display a unique profile have been underway. In this pursuit, we have identified one of the idenoisoquinoline derivatives, MJ-III-65 (NSC 706744; 6-[3-(2-hydroxyethyl)amino-1-propyl]-5,6 dihydro-2,3-dimethoxy-8,9-methylenedioxy-5,11-dioxo-11H-indeno[1,2 c]isoquinoline) with both similarities and differences from CPT. MJ-III-65 traps topoisomerase I (Top1) reversibly like CPT but with different DNA sequence preferences. Consistent with Top1 poisoning, protein-linked DNA breaks were detected in cells treated with MJ-III-65 at nanomolar concentrations. These MJ III-65-induced protein-linked DNA breaks were resistant to reversal after an hour of drug removal, compared with CPT, which completely reversed. Studies in human cells in culture found MJ-III-65 to be cytotoxic. Furthermore, limited cross resistance was observed in camptothecin-resistant cell lines. MJ-III-65 also exhibits antitumor activity in mouse tumor xenografts. PMID- 15531732 TI - Membrane structure modulation, protein kinase C alpha activation, and anticancer activity of minerval. AB - Most drugs currently used for human therapy interact with proteins, altering their activity to modulate the pathological cell physiology. In contrast, 2 hydroxy-9-cis-octadecenoic acid (Minerval) was designed to modify the lipid organization of the membrane. Its structure was deduced following the guidelines of the mechanism of action previously proposed by us for certain antitumor drugs. The antiproliferative activity of Minerval supports the above-mentioned hypothesis. This molecule augments the propensity of membrane lipids to organize into nonlamellar (hexagonal H(II)) phases, promoting the subsequent recruitment of protein kinase C (PKC) to the cell membrane. The binding of the enzyme to membranes was marked and significantly elevated by Minerval in model (liposomes) and cell (A549) membranes and in heart membranes from animals treated with this drug. In addition, Minerval induced increased PKCalpha expression (mRNA and protein levels) in A549 cells. This drug also induced PKC activation, which led to a p53-independent increase in p21(CIP) expression, followed by a decrease in the cellular concentrations of cyclins A, B, and D3 and cdk2. These molecular changes impaired the cell cycle progression of A549 cells. At the cellular and physiological level, administration of Minerval inhibited the growth of cancer cells and exerted antitumor effects in animal models of cancer without apparent histological toxicity. The present results support the potential use of Minerval and related compounds in the treatment of tumor pathologies. PMID- 15531733 TI - Alternatives to nitric oxide. AB - Inhaled nitric oxide (INO) is a selective pulmonary vasodilator that has the ability to produce vasodilation in the pulmonary vascular bed without causing it in the systemic circulation. This property of INO has made it a useful therapy in the management of both adult and paediatric patients with a variety of conditions associated with pulmonary hypertension (PH), with or without hypoxia. Toxicity, cost and negative-outcome studies have prompted a search for alternative agents. These include inhaled prostacyclin and alternative prostaglandin preparations such as inhaled iloprost, treprostinol and beraprost. The phospodiesterase inhibitors show real potential in the management of both acute and chronic forms of PH, and antagonists of endogenous pulmonary vasoconstrictors, such as endothelin and thromboxane, are being evaluated for the long-term treatment of conditions such as primary pulmonary hypertension. PMID- 15531734 TI - Adrenomedullin is highly expressed in blood monocytes associated with acute Kawasaki disease: a microarray gene expression study. AB - Kawasaki disease (KD) is an acute inflammatory disorder of children frequently associated with the development of coronary artery abnormalities. Although a great deal is known about inflammatory and immune responses in acute KD, the mechanisms linking the immune response to vascular changes are not known. To gain further insight into this process, we performed a microarray gene expression analysis on RNA isolated from the peripheral blood mononuclear cells of four patients with KD during both their acute and convalescent phases. Forty-seven genes of 7129 genes examined showed an increased expression in three or all four patients in the acute compared with the convalescent phase of KD. Fourteen of these genes were significantly (p < 0.05) up-regulated, including several inflammatory response genes (e.g. S-100 A9 protein) and also anti-inflammatory genes (e.g. TSG-6). Of greatest interest, the adrenomedullin (ADM) gene, known to be associated with coronary artery vasodilation, was up-regulated in the acute phase of KD (p = 0.024). Up-regulation of ADM in the acute phase of KD was confirmed in peripheral blood mononuclear cells of 11 additional KD patients by reverse transcriptase-PCR (p < 0.01). Isolated blood monocytes but not lymphocytes were demonstrated by real-time PCR to have increased ADM mRNA (p = 0.01). Plasma ADM protein level in 32 additional KD patients was also confirmed to be higher in acute KD compared with convalescent KD (p < 0.032). It is interesting that from microarray results, other molecules known to be associated with coronary dilation, including nitric oxide, prostacyclin, acetylcholine, bradykinin, substance P, and serotonin, were not elevated in acute KD. Our current study suggests that ADM-expressing monocytes that infiltrate the coronary vascular wall may be the cause of coronary dilation in the acute phase of KD. PMID- 15531735 TI - Sildenafil citrate (viagra) induces cardioprotective effects after ischemia/reperfusion injury in infant rabbits. AB - Infants undergoing surgery for congenital heart disease are at risk for myocardial ischemia during cardiopulmonary bypass, circulatory arrest, or low flow states. The purpose of this study was to demonstrate the effects of sildenafil, a selective phosphodiesterase-5 (PDE-5) inhibitor on myocardial functional improvement and infarct size reduction during ischemia/reperfusion injury in infant rabbits. Infant rabbits (aged 8 wk) were treated with sildenafil citrate (0.7 mg/kg i.v.) or normal saline 30 min before sustained ischemia for 30 min and reperfusion for 3 h. Transesophageal echocardiography (TEE) was used to assess left ventricular cardiac output (LVCO) and aortic velocity time integral (VTI). After ischemia/reperfusion, risk area was demarcated by Evan's blue dye and infarct size determined by computer morphometry of triphenyltetrazolium chloride-stained sections. The sildenafil-treated group had preservation and elevation in LVCO (143% of baseline, p < 0.05) and an elevated aortic VTI (145% of baseline, p < 0.05) after 30 min of ischemia compared with the control group LVCO (72% of baseline, p < 0.05) and aortic VTI (73% of baseline, p < 0.05). This is a statistically significant increase in LVCO and aortic VTI in the sildenafil group compared with controls (n = 6/group, p < 0.05). The sildenafil-treated group had significant reduction in infarct size (15.5 +/- 1.2 versus 33 +/- 2.3 in the saline group, % risk area, mean +/- SEM, n = 10-15/group, p < 0.05). For the first time, we have shown that sildenafil citrate promotes myocardial protection in infant rabbits as evidenced by postischemic preservation and elevation in LVCO and aortic VTI and reduction in infarct size. PMID- 15531736 TI - Thymic function and impaired maintenance of peripheral T cell populations in children with congenital heart disease and surgical thymectomy. AB - The thymus begins involution in childhood and historically it was thought to be nonfunctional by adulthood, thus presenting no contraindication to the routine practice of thymectomy during cardiothoracic surgery. More recent data suggest, however, that the thymus remains active into adulthood and is responsible for the low-level production of normal T cells. We hypothesize, therefore, that incidental thymectomy during cardiothoracic surgery in infancy causes long-term changes in the cellular immune system. To investigate this hypothesis, we quantified peripheral T-cell subsets and T-cell recombination excision circles in children with congenital heart disease to measure the impact of cardiothoracic surgical procedures and thymectomy performed during a period of immunologic development. We found that cardiothoracic surgical procedures, especially if they include thymectomy, impair T-cell production and produce long-term decreases in total lymphocyte count and CD4(+) and CD8(+) T-cell subsets, suggesting that long term maintenance of lymphocyte populations is disturbed. PMID- 15531737 TI - Systolic and diastolic ventricular function assessed by pressure-volume loops in the stage 21 venous clipped chick embryo. AB - Cardiac pressure-volume relations enable quantification of intrinsic ventricular diastolic and systolic properties independent of loading conditions. The use of pressure-volume loop analysis in early stages of development could contribute to a better understanding of the relationship between hemodynamics and cardiac morphogenesis. The venous clip model is an intervention model for the chick embryo in which permanent obstruction of the right lateral vitelline vein temporarily reduces the mechanical load on the embryonic myocardium and induces a spectrum of outflow tract anomalies. We used pressure-volume loop analysis of the embryonic chick heart at stage 21 (3.5 d of incubation) to investigate whether the development of ventricular function is affected by venous clipping at stage 17, compared with normal control embryos. Steady state hemodynamic parameters demonstrated no significant differences between the venous clipped and control embryos. However, analysis of pressure-volume relations showed a significantly lower end-systolic elastance in the clipped embryos (slope of the end-systolic pressure-volume relation: 5.68 +/- 0.85 versus 11.76 +/- 2.70 mm Hg/microL, p < 0.05), indicating reduced contractility. Diastolic stiffness tended to be increased in the clipped embryos (slope of end-diastolic pressure-volume relation: 2.74 +/- 0.56 versus 1.67 +/- 0.21, p = 0.103), but the difference did not reach statistical significance. The results of the pressure-volume loop analysis show that 1 d after venous obstruction, development of ventricular function is affected, with reduced contractility. Pressure-volume analysis may be applied in the chick embryo and is a sensitive technique to detect subtle alterations in ventricular function. PMID- 15531738 TI - PPAR-gamma2 Pro12Ala variant is associated with greater insulin sensitivity in childhood obesity. AB - Several genetic variants of peroxisome proliferator-activated receptor-gamma2 (PPAR-gamma2), a molecule known to be involved in transcription of target genes, have been identified. Pro12Ala, a missense mutation in exon 2 of the gene, is highly prevalent in Caucasian populations. Conflicting conclusions about the association between this mutation and complex traits such as obesity, insulin sensitivity, and T2DM have been reported. We have investigated the association of PPAR-gamma2 Pro12Ala polymorphism with measures of insulin sensitivity in a population of Italian obese children (n = 200; mean age, 10.38 +/- 2.8 y) in whom clinical and biochemical analyses were performed. To estimate the insulin sensitivity status, the homeostasis model assessment of insulin resistance (HOMA IR) was calculated in all subjects. The effect of the Pro12Ala polymorphism on quantitative variables was tested using multiple linear regression analysis. The frequency of Ala carriers was 17%, similar to that reported in other adult Caucasian populations. The X12Ala (either Pro12Ala or Ala12Ala) genotype was associated with significantly lower fasting insulin levels compared with Pro/Pro (p = 0.008). Consistent with this finding, significantly lower HOMA-IR was observed in X12Ala carriers (p = 0.023). In conclusion, our observations demonstrate that the X12Ala variant is significantly associated with greater insulin sensitivity in childhood obesity. Because obesity is one of the most important risk factors for cardiovascular diseases and type 2 diabetes, obese children, who are presumably at a higher risk, may be protected from these diseases by the phenotypic effect of the Ala 12 allele on insulin resistance. PMID- 15531739 TI - Identification of pressure passive cerebral perfusion and its mediators after infant cardiac surgery. AB - Cerebrovascular pressure autoregulation (CPA) regulates cerebral blood flow (CBF) in relation to changes in mean arterial blood pressure (MAP). Identification of a pressure-passive cerebral perfusion and the potentially modifiable physiologic factors underlying it has been difficult to achieve in sick infants. We previously validated the near-infrared spectroscopy-derived hemoglobin difference (HbD) signal (cerebral oxyhemoglobin - deoxyhemoglobin) as a reliable measure of changes in CBF in animal models. We now sought to determine whether continuous measurements of DeltaHbD would correlate to middle cerebral artery flow velocity (CBFV), allow identification and quantification of pressure-passive state, and help to delineate potentially modifiable factors. We enrolled 43 infants (2 d to 7 mo old) who were undergoing open cardiac surgery and cardiopulmonary bypass. At 6 and 20 h after surgery, we measured changes in HbD, CBFV (by transcranial Doppler), and MAP at different end-tidal CO(2) levels. We assigned a pressure passive index (PPI) to each study on the basis of the relative duration of significant coherence between DeltaMAP and DeltaHbD. We found a significant relationship between DeltaHbD and DeltaCBFV at both time points. At 6 h after surgery, we showed high concordance (coherence > 0.5; PPI > or = 41%) between DeltaMAP and DeltaHbD, consistent with disturbed CPA in 13% of infants. End-tidal CO(2) values > or = 40 mm Hg and higher MAP variability both were associated with increased odds (p < 0.001) of autoregulatory failure. This approach provides a means to identify and quantify disturbances of CPA. High CO(2) levels and fluctuating MAP are two important preventable factors associated with disturbed CPA. PMID- 15531740 TI - Where does the developing brain obtain its docosahexaenoic acid? Relative contributions of dietary alpha-linolenic acid, docosahexaenoic acid, and body stores in the developing rat. AB - Docosahexaenoic acid (DHA), a 22-carbon, highly unsaturated, n-3 fatty acid, is important for optimal nervous system function. In this study, designed to quantify how preformed dietary DHA regulates metabolic pathways in vivo, 8-d-old rat pups were divided into four groups and fed artificial rat milk diets. One group was fed formula with deuterium-labeled LNA (d5-LNA) as the only source of n 3 fatty acids, and a second group was fed formula that contained d5-LNA and unlabeled DHA. Two additional groups were dam-reared to permit analysis of fatty acyl pool sizes at postnatal days 8 and 28. The dams were fed a diet that contained 3% unlabeled LNA. DHA in brain and liver was analyzed. Our study demonstrated that preformed DHA in the diet markedly decreased the amount of biosynthesized DHA that accumulated in the brain and the liver. Surprisingly, 40% of the DHA that was newly acquired during this period in the "LNA" group was unlabeled. Because there were no unlabeled n-3 fatty acids in this diet, this DHA must have been derived from body stores of n-3 fatty acids. Thus, body stores can be a significant source of brain DHA in animals that are fed LNA as the only source of n-3 fatty acids. PMID- 15531741 TI - Evolving therapeutic strategies for Duchenne muscular dystrophy: targeting downstream events. AB - Duchenne muscular dystrophy (DMD) is a progressive, lethal, muscle wasting disease that affects 1 of 3500 boys born worldwide. The disease results from mutation of the dystrophin gene that encodes a cytoskeletal protein associated with the muscle cell membrane. Although gene therapy will likely provide the cure for DMD, it remains on the distant horizon, emphasizing the need for more rapid development of palliative treatments that build on improved understanding of the complex pathology of dystrophin deficiency. In this review, we have focused on therapeutic strategies that target downstream events in the pathologic progression of DMD. Much of this work has been developed initially using the dystrophin-deficient mdx mouse to explore basic features of the pathophysiology of dystrophin deficiency and to test potential therapeutic interventions to slow, reverse, or compensate for functional losses that occur in muscular dystrophy. In some cases, the initial findings in the mdx model have led to clinical treatments for DMD boys that have produced improvements in muscle function and quality of life. Many of these investigations have concerned interventions that can affect protein balance in muscle, by inhibiting specific proteases implicated in the DMD pathology, or by providing anabolic factors or depleting catabolic factors that can contribute to muscle wasting. Other investigations have exploited the use of anti-inflammatory agents that can reduce the contribution of leukocytes to promoting secondary damage to dystrophic muscle. A third general strategy is designed to increase the regenerative capacity of dystrophic muscle and thereby help retain functional muscle mass. Each of these general approaches to slowing the pathology of dystrophin deficiency has yielded encouragement and suggests that targeting downstream events in dystrophinopathy can yield worthwhile, functional improvements in DMD. PMID- 15531742 TI - The influence of cortical maturation on the BOLD response: an fMRI study of visual cortex in children. AB - We performed blood oxygenation level-dependent (BOLD) functional MR imaging in 11 children younger than 5 y of age and 10 children older than 5 y of age. All but five of the children in the older age group were tested under light anesthesia. We examined the cerebral oxidative metabolism (CMRO(2)) associated with the processing of a flashed and a reversing checkerboard stimulus. These stimuli had been shown in a previous study to induce identical vascular responses. The reversing checkerboard activated twice the neuronal population of the flashed checkerboard, doubling the CMRO(2) associated with it. We compared the extent of activation for the positive BOLD response and found that it did not differ between the different age groups. We estimated the oxidative metabolism by examining the change in the local deoxyhemoglobin (HbR) concentration using Delta R2*. Because both stimuli induced the same vascular response, any increase in oxygen requirement would have to be met by the identical blood volume. Increasing CMRO(2) will therefore result in an increase in the oxygen extraction fraction (OEF), which raises the local HbR concentration. In the younger children, both checkerboard stimuli produced identical, high HbR concentrations. In the older children, the HbR concentration to the flashed stimulus was significantly lower than to the reversing stimulus. We conclude that, for identical stimuli, the oxidative energy requirement associated with the cortical processing is higher in young children than in older children because the presence of superfluous synaptic connections in the immature visual system activates a larger neuronal population. PMID- 15531743 TI - Fetal alcohol exposure impairs alveolar macrophage function via decreased glutathione availability. AB - Immature function of the alveolar macrophage increases the risk of pulmonary infections in premature newborns. In utero alcohol increases fetal systemic oxidative stress. Because the premature lung is deficient in glutathione (GSH), we hypothesized that chronic in utero alcohol (ethanol) exposure exacerbates the oxidative stress within the developing lung, thereby impairing alveolar macrophage function. Additionally, we evaluated the effects of in vivo and in vitro GSH availability on ethanol-exposed macrophage function. Using a guinea pig model of chronic in utero ethanol exposure, fetal epithelial lining fluid (ELF) and alveolar macrophage GSH were decreased with increased markers of oxidative stress. Ethanol-exposed macrophage exhibited impaired phagocytosis and increased apoptosis compared with gestational control. When the GSH precursor S-adenosyl methionine (SAM) was added to the maternal drinking water containing ethanol, fetal ELF and macrophage GSH were maintained and ELF oxidative stress diminished. In vivo maternal SAM therapy maintained macrophage phagocytosis and decreased apoptosis. In vitro GSH supplements also improved phagocytosis and viability in both premature and ethanol-exposed macrophage. This suggested that in utero ethanol impaired premature macrophage function and viability via decreased GSH availability. Furthermore, GSH supplementation during and after ethanol exposure improved fetal macrophage function and viability. These results add a new dimension to the detrimental effects of fetal alcohol exposure on the developing alveolar macrophage, raising the possibility of GSH therapy to augment premature alveolar macrophage function. PMID- 15531744 TI - Expression and secretion of cathelicidin antimicrobial peptides in murine mammary glands and human milk. AB - Mammalian milk possesses inherent antimicrobial properties that have been attributed to several diverse molecules. Recently, antimicrobial peptides that belong to the cathelicidin gene family have been found to be important to the mammalian immune response. This antimicrobial is expressed in several tissues and increased in neonatal skin, possibly to compensate for an immature adaptive immune response. We hypothesized that the mammary gland could produce and secrete cathelicidin onto the epithelial surface and into milk. Human cathelicidin hCAP18/LL-37 mRNA was detected in human milk cells by PCR. Quantitative real-time PCR demonstrated an increase in relative expression levels at 30 and 60 d after parturition. Immunohistochemistry of mouse breast tissue identified the murine cathelicidin-related antimicrobial peptide in lobuloacinar and ductules. Western blot analysis of human milk showed that LL-37 was secreted and present in the mature peptide form. The antimicrobial activity of LL-37 against Staphylococcus aureus, group A Streptococcus, and enteroinvasive Escherichia coli O29 in the human milk ionic environment was confirmed by solution colony-forming assay using synthetic peptide. These results indicate that cathelicidin is secreted in mammary gland and human milk, has antimicrobial activity against both Gram positive and Gram-negative bacteria, and can contribute to the anti-infectious properties of milk. PMID- 15531745 TI - Sleep-dependent changes in the coupling between heart period and arterial pressure in newborn lambs. AB - This study assessed whether sleep-dependent changes in the relationship between heart period (HP) and mean arterial pressure (MAP) occur in newborn life. Electrodes for electrocorticographic, electromyographic, and electrooculographic monitoring and an arterial catheter for blood pressure recordings were implanted in 11 newborn lambs. HP and MAP beat-to-beat values were computed from 120-s blood pressure recordings during quiet wakefulness, active sleep, and quiet sleep. For each recording, the time shift at which the maximum of the HP versus MAP cross-correlation function was attained was identified. For each lamb and wake-sleep state, an average correlation coefficient was then computed corresponding to the median value of such time shifts. The maximum of the cross correlation function was attained with HP lagging behind MAP. The corresponding mean correlation coefficient was significantly higher in quiet sleep (0.51 +/- 0.05) than either in quiet wakefulness (0.31 +/- 0.05) or in active sleep (0.29 +/- 0.03). Sleep-related differences in the correlation between HP and MAP were maintained after HP and MAP data were low-pass filtered at 0.3 Hz to remove their fast ventilatory oscillations. In conclusion, data indicate that the relationship between spontaneous fluctuations in HP and those in MAP is sleep-state dependent in newborn lambs. A positive HP versus MAP correlation with HP lagging behind MAP is consistent with baroreflex control of HP. Heart rhythm thus may be more tightly controlled by the baroreceptor reflex and less dependent on central autonomic commands in quiet sleep than either in quiet wakefulness or in active sleep. PMID- 15531746 TI - Angiotensin-converting enzyme inhibition modulates mitogen-activated protein kinase family expressions in the neonatal rat kidney. AB - Among the mitogen-activated protein kinase (MAPK) family members, extracellular signal-regulated kinase (ERK) promotes cell proliferation or differentiation, whereas c-jun N terminal kinase (JNK) and p38 MAPK are thought to inhibit cell growth and induce apoptosis. The MAPK family may plays some role during kidney development, when large-scale proliferation and apoptosis have been observed to occur. Also, in this period, the renin-angiotensin system is markedly activated. We have demonstrated that angiotensin-converting enzyme inhibition in the developing rat kidney increases apoptosis and decreases cell proliferation, which may account for renal growth impairment. The aim of this study, therefore, was to examine the relationship between the MAPK family and renin-angiotensin system during neonatal renal development. Newborn rat pups were treated with enalapril (30 mg . kg(-1) . d(-1)) or normal saline for 7 d. Right kidneys of both groups were selected for immunohistochemical stains of MAPKs and activating transcription factor-2 (ATF-2), and left kidneys were selected for reverse transcriptase-PCR and immunoblot analysis of MAPKs, phospho-MAPKs, and ATF-2. To determine whether apoptosis is involved in the same tubules that highly expressed JNK and p38, we performed terminal deoxynucleotide transferase-mediated nick-end labeling stain for apoptotic cells and immunohistochemical stains for JNK-2, p38, and ATF-2 expression in the serial sections from the same kidney of the enalapril treated group. In the enalapril-treated group, JNK-2, p38, phospho-JNK-2, phospho p38, and ATF-2 protein expressions were significantly increased, and their immunoactivities were strongly detected in the proximal tubular epithelial cells in the cortex, compared with the control group. Especially JNK-2 and p38 expressions were highly activated and were spatially in accordance with the occurrence of apoptosis. ERK1/2 and phospho-ERK expressions were not changed by enalapril. These results suggest that the expressions of the MAPK family are modulated by angiotensin-converting enzyme inhibition in the developing kidney. JNK and p38 may be implicated to participate in angiotensin II-related intracellular signaling pathways of renal apoptosis in the developing kidney. PMID- 15531747 TI - Abnormal RNA processing and altered expression of serin-rich proteins in minimal change nephrotic syndrome. AB - Mechanisms underlying the pathophysiology of minimal-change nephrotic syndrome (MCNS), the most frequent glomerular disease in children, remain elusive, but recent findings argue for a T cell dysfunction. Starting from a differential cDNA library from T cells of a patient under relapse and remission, we identified 16 transcripts specific for MCNS. All of these transcripts that were selectively up regulated during the relapse phase of the disease were generated by alternative splicing of known genes. This abnormal RNA expression was associated with a down regulation of serin-rich protein 75 and serin-rich protein 40, two proteins involved in mRNA splicing. Taken together, these data suggest that T cell dysfunction in MCNS is associated with abnormal mRNA splicing. PMID- 15531748 TI - Endothelin-1 decreases endothelial NOS expression and activity through ETA receptor-mediated generation of hydrogen peroxide. AB - Similar to infants born with persistent pulmonary hypertension of the newborn (PPHN), there is an increase in circulating endothelin-1 (ET-1) and decreased endothelial nitric oxide synthase (eNOS) gene expression in an ovine model of PPHN. These abnormalities lead to vasoconstriction and vascular remodeling. Our previous studies have demonstrated that reactive oxygen species (ROS) levels are elevated in the pulmonary arteries from PPHN lambs and that ET-1 increases ROS production in pulmonary arterial smooth muscle cells (PASMC) in culture. Thus the objective of this study was to determine whether there was a feedback mechanism between the ET-1-mediated increase in ROS in fetal PASMC (FPASMC) and a decrease in eNOS gene expression in fetal pulmonary arterial endothelial cells (FPAEC). Our results indicate that ET-1 increased H2O2 levels in FPASMC in an endothelin A receptor-dependent fashion. This was observed in both FPASMC monoculture and in cocultures of FPASMC and FPAEC. Conversely, ET-1 decreased H2O2 levels in FPAEC monoculture in an endothelin B receptor-dependent fashion. Furthermore, ET-1 decreased eNOS promoter activity by 40% in FPAEC in coculture with FPASMC. Promoter activity was restored in the presence of catalase. In FPAEC in monoculture treated with 0-100 microM H2O2, 12 microM had no effect on eNOS promoter activity, but it increased eNOS protein levels by 50%. However, at 100 microM, H2O2 decreased eNOS promoter activity and protein levels in FPAEC by 79 and 40%, respectively. These data suggest a role for smooth muscle cell-derived H2O2 in ET-1-mediated downregulation of eNOS expression in children born with PPHN. PMID- 15531749 TI - Metalloproteinases mediate mucin 5AC expression by epidermal growth factor receptor activation. AB - Chronic obstructive pulmonary disease is marked by alveolar enlargement and excess production of airway mucus. Acrolein, a component of cigarette smoke, increases mucin 5AC (MUC5AC), a prevalent airway mucin in NCI-H292 cells by transcriptional activation, but the signal transduction pathways involved in acrolein-induced MUC5AC expression are unknown. Acrolein depleted cellular glutathione at doses of 10 muM or greater, higher than those sufficient (0.03 muM) to increase MUC5AC mRNA, suggesting that MUC5AC expression was independent of oxidative stress. In contrast, acrolein increased MUC5AC mRNA levels by phosphorylating epidermal growth factor receptor (EGFR) and mitogen-activated protein kinase 3/2, or MAPK 3/2(ERK1/2). Pretreating the cells with an EGFR neutralizing antibody, or a metalloproteinase inhibitor, decreased the acrolein induced MUC5AC mRNA increase. Small, interfering RNA directed against ADAM17 or MMP9 inhibited the acrolein-induced MUC5AC mRNA increase. Acrolein increased the release and subsequent activation of pro-MMP9. Acrolein increased MMP9 and decreased tissue inhibitor of metalloproteinase 3 (TIMP3), an endogenous inhibitor of ADAM17, transcripts. Together, these data suggest that acrolein induces MUC5AC expression via an initial ligand-dependent activation of EGFR mediated by ADAM17 and MMP9. In addition, a prolonged effect of acrolein may be mediated by altering MMP9 and TIMP3 transcription. PMID- 15531750 TI - Ventilation inhomogeneities in relation to standard lung function in patients with cystic fibrosis. AB - Based on serial lung function measurements performed in 142 children (68 males; 74 females) with cystic fibrosis (CF), prospectively evaluated over an age range of 6 to 20 years, we attempted to determine whether the lung clearance index (LCI) as a measure of ventilation inhomogeneities could be a discriminating factor of disease progression. Annual follow-up lung function measurements featuring FRC determined by whole-body plethysmography and multibreath nitrogen washouts, effective specific airway resistance, flow-volume curves, LCI, and gas exchange characteristics were analyzed by linear mixed-model analysis and Kaplan Meier statistics. The earliest occurring and strongest factor of progression was the LCI, followed by maximal expiratory flow (MEF(50)) and FRC determined by plethysmography (p < 0.0001). Associations between onset of chronic Pseudomonas aeruginosa infection and CF transmembrane conductance regulator (CFTR) genotype with FEV(1) (p = 0.027) and FVC (p = 0.007) were identified. The study shows that the LCI predicts earlier in life and represented much better functional progression than FEV(1). Moreover, there is no single functional predictor of progression in CF, but aside from risk factors, such as onset of chronic P. aeruginosa infection and genotype, pulmonary hyperinflation, airway obstruction, and ventilation inhomogeneities are important pathophysiologic processes that should be evaluated concomitantly as determinants of lung progression in CF. PMID- 15531751 TI - Microsomal epoxide hydrolase, endotoxin, and lung function decline in cotton textile workers. AB - Occupational exposure to endotoxin in organic dust may induce lung function decline. Microsomal epoxide hydrolase (mEH) detoxifies reactive oxygen species generated by endotoxin exposure, and polymorphisms of the mEH gene are associated with altered enzyme activity. We investigated the associations between mEH polymorphisms, endotoxin exposure, and lung function decline in a 20-year prospective study of 265 workers exposed to endotoxin and 234 control subjects. mEH Tyr113His and His139Arg polymorphisms were genotyped by the 5' nuclease assay, and data were analyzed using multivariate linear regression models, adjusting for important covariates. Overall, the annual decline rate of FEV1 was 29.47 ml during the 20-year follow-up. Endotoxin exposure was associated with faster lung function decline among genotypes associated with slower enzyme activity: estimates (SE) of annual FEV1 decline rates for endotoxin exposure were -2.33 (2.07), -2.81 (1.66), and -6.73 (2.83) ml for Tyr/Tyr, Tyr/His, and His/His genotype groups, respectively, for the Tyr113His polymorphism; and -1.82 (2.58) and -4.27 (1.33) ml for Arg/Arg + His/Arg and His/His genotypes, respectively, for the His139Arg polymorphism. We conclude that mEH polymorphisms modify the association between occupational endotoxin exposure and longitudinal lung function decline. PMID- 15531752 TI - Systemic inflammatory response and progression to severe sepsis in critically ill infected patients. AB - RATIONALE: The systemic inflammatory response syndrome has low specificity to identify infected patients at risk of worsening to severe sepsis or shock. OBJECTIVE: To examine the incidence of and risk factors for worsening sepsis in infected patients. METHODS: A 1-year inception cohort study in 28 intensive care units of patients (n = 1,531) having a first episode of infection on admission or during the stay. MEASUREMENTS AND MAIN RESULTS: The cumulative incidence of progression to severe sepsis or shock was 20% and 24% at Days 10 and 30, respectively. Variables independently associated (hazard ratio [HR]) with worsening sepsis included: temperature higher than 38.2 degrees C (1.6), heart rate greater than 120/minute (1.3), systolic blood pressure higher than 110 mm Hg (1.5), platelets higher than 150 x 109/L (1.5), serum sodium higher than 145 mmol/L (1.5), bilirubin higher than 30 mumol/L (1.3), mechanical ventilation (1.5), and five variables characterizing infection (pneumonia [HR 1.5], peritonitis [1.5], primary bacteremia [1.8], and infection with gram-positive cocci [1.3] or aerobic gram-negative bacilli [1.4]). The 12 weighted variables were included in a score (Risk of Infection to Severe Sepsis and Shock Score, range 0-49), summarized in four classes of "low" (score 0-8) and "moderate" (8.5 16) risk (9% and 17% probability of worsening, respectively), and of "high" (16.5 24) and "very high" (score > 24) risk (31% and 55% probability, respectively). CONCLUSIONS: One of four patients presenting with infection/sepsis worsen to severe sepsis or shock. A score estimating this risk, using objectively defined criteria for systemic inflammatory response syndrome, could be used by physicians to stratify patients for clinical management and to test new interventions. PMID- 15531753 TI - Efficacy of fluticasone propionate on lung function and symptoms in wheezy infants. AB - The role of inhaled corticosteroids in the treatment of recurrent or persistent wheeze in infancy remains unclear. We evaluated the effect of 3 months of treatment with inhaled fluticasone propionate, 200 microg daily (FP200), on lung function and symptom scores in wheezy infants. Moreover, we evaluated whether infants with atopy and/or eczema respond better to FP200 as compared with non atopic infants. Forced expiratory flow (Vmax(FRC)) was measured at baseline and after treatment. Sixty-five infants were randomized to receive FP200 or placebo, and 62 infants (mean age, 11.3 months) completed the study. Mean Vmax(FRC), expressed as a Z score, was significantly below normal at baseline and after treatment in both groups. The change from baseline of Vmax(FRC) was not different between the two treatment arms. After 6 weeks of treatment, and not after 13 weeks, the FP200 group had a significantly higher percentage of symptom-free days and a significant reduction in mean daily cough score compared with placebo. Separate analysis of treatment effect in infants with atopy or eczema showed no effect modification. We conclude that in wheezy infants, after 3 months of treatment with fluticasone, there was no improvement in lung function and no reduction in respiratory symptoms compared with placebo. PMID- 15531754 TI - Antioxidant intake in pregnancy in relation to wheeze and eczema in the first two years of life. AB - Two thousand women were recruited for a prospective investigation of the influence of maternal antioxidant intake in pregnancy on the development of asthma and eczema in children. A food frequency questionnaire was used to characterize diet during pregnancy and blood antioxidant levels were measured. Postal questionnaires were used to follow up the 1,924 singleton children born to the cohort at 6, 12, and 24 months of age. There were no associations between maternal antioxidant intake and wheezing symptoms and eczema in the children's first year. In the children's second year, maternal vitamin E intake during pregnancy was negatively associated with wheeze in the absence of a "cold" (p for trend 0.010) and, in children whose mothers were atopic, there was a negative association between maternal vitamin E intake and childhood eczema (p for trend 0.024). Maternal vitamin C intake during pregnancy was positively associated with "ever wheeze" and eczema during the children's second year. This study suggests that maternal dietary antioxidant intakes during pregnancy may modify the risks of developing wheeze and eczema during early childhood. Further follow up of the cohort will determine whether maternal diet during pregnancy is associated with asthma and atopic disease in later childhood. PMID- 15531755 TI - Understanding pulmonary gas exchange: ventilation-perfusion relationships. AB - This essay looks at the historical significance of four APS classic papers that are freely available online: Fenn WO, Rahn H, and OTIS AB. A theoretical study of the composition of the alveolar air at altitude. Am J Physiol 146: 637-653. 1946 (http://ajplegacy.physiology.org/cgi/reprint/146/5/637). Rahn H. A concept of mean alveolar air and the ventilation-bloodflow relationships during pulmonary gas exchange. Am J Physiol 158: 21-30, 1949 (http://ajplegacy.physiology.org/cgi/reprint/158/1/21)). Riley RL. And Cournand A. "Ideal" Alveolar air and the analysis of ventilation-perfusion relationships in the lungs. J Appl Physiol 1: 825-847. 1949 (http://jap.physiology.org/cgi/reprint/1/12/825). Riley RL. And Cournand A. Analysis of factors affecting partial pressures of oxygen and carbon dioxide in gas and blood of lungs: theory. J Appl Physiol 4: 77-101. 1951 (http://jap.physiology.org/cgi/reprint/4/2/77). PMID- 15531756 TI - The promise and perils of exhaled breath condensates. AB - The exhaled breath condensate (EBC) approach provides a convenient and noninvasive approach for sampling the pulmonary epithelial lining fluid (ELF). Increased EBC concentrations of more than a dozen inflammatory markers and hydrogen ions have been reported in lung diseases associated with inflammation. However, the usefulness of EBC is compromised by uncertainties concerning the sources of the EBC droplets and by the extreme and variable dilution of ELF droplets with condensed water vapor ( approximately 20,000-fold). Reported increases in EBC concentrations may reflect proportionate increases in the total volume rather than the concentration of ELF droplets in the collected samples. Conclusions regarding ELF concentrations can only be made if this dilution is estimated with a dilutional indicator (e.g., conductivity of lyophilized EBC). In normal EBC samples, pH is effectively set by oral contamination with NH(3), and EBC pH cannot provide reliable information regarding ELF pH in normal subjects. Acidification of EBC observed in asthma and other conditions may reflect acidification of ELF, decreases in NH(3) added to the EBC, and/or the presence of gastric droplets in the EBC. PMID- 15531757 TI - Integrated control of lung fluid balance. AB - This review summarizes the highlights of the EB2004 symposium that dealt with the integrated aspects of the lung fluid balance. It is apparent that maintenance of lung fluid balance requires the proper functioning of vascular endothelial and alveolar epithelial barriers. Under physiological conditions, the transcytotic pathway requiring repeated fission-fusion events of the caveolar membrane with other caveolae solely transports albumin. Caveolin-1, which forms caveolae, and albumin-binding proteins play a central role in signaling the transcytosis of albumin. Signals responsible for increasing endothelial permeability in lung microvessels in response to inflammatory mediators were also described. These studies in gene knockout mouse models revealed the importance of Ca(2+) signaling via store-operated transient receptor channel 4 and the activation of endothelial myosin light chain kinase isoform in mediating the increase in microvessel permeability. Increases in the cytosolic Ca(2+) in situ in microvessel endothelia can occur by mitochondria-dependent as well as mitochondria-independent pathways (such as the endoplasmic reticulum). Both these pathways, by triggering endothelial cell activation, may result in lung microvascular injury. The resolution of alveolar edema, requiring clearance of fluid from the air space, is another area of intense investigation in animal models. Although beta-adrenergic agonists can activate alveolar fluid clearance, signaling pathways regulating these events in intact alveoli remain to be established. Development of mouse models in which the function of regulatory proteins (identified in cell culture studies) can be systematically analyzed will provide a better and more integrated picture of lung fluid balance. In vivo veritas! PMID- 15531758 TI - FGF-10 induces SP-C and Bmp4 and regulates proximal-distal patterning in embryonic tracheal epithelium. AB - The induction, growth, and differentiation of epithelial lung buds are regulated by the interaction of signals between the lung epithelium and its surrounding mesenchyme. Fibroblast growth factor-10 (FGF-10), which is expressed in the mesenchyme near the distal tips, and bone morphogenetic protein 4 (BMP4), which is expressed in the most distal regions of the epithelium, are important molecules in lung morphogenesis. In the present study, we used two in vitro systems to examine the induction, growth, and differentiation of lung epithelium. Transfilter cultures were used to determine the effect of diffusible factors from the distal lung mesenchyme (LgM) on epithelial branching, and FGF-10 bead cultures were used to ascertain the effect of a high local concentration of a single diffusible molecule on the epithelium. Embryonic tracheal epithelium (TrE) was induced to grow in both culture systems and to express the distal epithelial marker surfactant protein C at the tips nearest the diffusible protein source. TrE cultured on the opposite side of a filter to LgM branched in a pattern resembling intact lungs, whereas TrE cultured in apposition to an FGF-10 bead resembled a single elongating epithelial bud. Examination of the role of BMP4 on lung bud morphogenesis revealed that BMP4 signaling suppressed expression of the proximal epithelial genes Ccsp and Foxj1 in both types of culture and upregulated the expression of Sprouty 2 in TrE cultured with an FGF-10 bead. Antagonizing BMP signaling with Noggin, however, increased expression of both Ccsp and Foxj1. PMID- 15531759 TI - Decreased expression of uteroglobin-related protein 1 in inflamed mouse airways is mediated by IL-9. AB - Uteroglobin-related protein 1 (UGRP1) is a secretory protein, highly expressed in epithelial cells of airways. Although an involvement of UGRP1 in the pathogenesis of asthma has been suggested, its function in airways remains unclear. In the present study, a relationship between airway inflammation, UGRP1 expression, and interleukin-9 (IL-9), an asthma candidate gene, was evaluated by using a murine model of allergic bronchial asthma. A severe airway inflammation accompanied by airway eosinophilia and elevation of IL-9 in bronchoalveolar lavage (BAL) fluids was observed after ovalbumin (OVA) challenge to OVA-sensitized mice. In this animal model of airway inflammation, lung Ugrp1 mRNA expression was greatly decreased compared with control mice. A significant inverse correlation between lung Ugrp1 mRNA levels and IL-9 levels in BAL fluid was demonstrated by regression analysis (r = 0.616, P = 0.023). Immunohistochemical analysis revealed a distinct localization of UGRP1 in airway epithelial cells of control mice, whereas UGRP1 staining was patchy and faint in inflamed airways. Intranasal administration of IL-9 to naive mice decreased the level of Ugrp1 expression in lungs. These findings suggest that UGRP1 is downregulated in inflamed airways, such as allergic asthmatics, and IL-9 might be an important mediator for modulating UGRP1 expression. PMID- 15531760 TI - Degradation of MyoD mediated by the SCF (MAFbx) ubiquitin ligase. AB - MyoD controls myoblast identity and differentiation and is required for myogenic stem cell function in adult skeletal muscle. MyoD is degraded by the ubiquitin proteasome pathway mediated by different E3 ubiquitin ligases not identified as yet. Here we report that MyoD interacts with Atrogin-1/MAFbx (MAFbx), a striated muscle-specific E3 ubiquitin ligase dramatically up-regulated in atrophying muscle. A core LXXLL motif sequence in MyoD is necessary for binding to MAFbx. MAFbx associates with MyoD through an inverted LXXLL motif located in a series of helical leucine-charged residue-rich domains. Mutation in the LXXLL core motif represses ubiquitination and degradation of MyoD induced by MAFbx. Overexpression of MAFbx suppresses MyoD-induced differentiation and inhibits myotube formation. Finally the purified recombinant SCF(MAFbx) complex (SCF, Skp1, Cdc53/Cullin 1, F box protein) mediated MyoD ubiquitination in vitro in a lysine-dependent pathway. Mutation of the lysine 133 in MyoD prevented its ubiquitination by the recombinant SCF(MAFbx) complex. These observations thus demonstrated that MAFbx functions in ubiquitinating MyoD via a sequence found in transcriptional coactivators. These transcriptional coactivators mediate the binding to liganded nuclear receptors. We also identified a novel protein-protein interaction module not yet identified in F-box proteins. MAFbx may play an important role in the course of muscle differentiation by determining the abundance of MyoD. PMID- 15531761 TI - Differential regulation of chemokine expression by peroxisome proliferator activated receptor gamma agonists: interactions with glucocorticoids and beta2 agonists. AB - Chemokine-mediated inflammatory cell infiltration is a hallmark of asthma. We recently demonstrated that glucocorticoids and beta(2)-agonists additively or synergistically suppress tumor necrosis factor-alpha (TNFalpha)-induced production of chemokines eotaxin and interleukin-8 (IL-8), respectively, in human airway smooth muscle (HASM) cells, which may partly explain their combined benefits in asthma. Peroxisome proliferator-activated receptors (PPARs) also modulate inflammatory gene expression. We reported here that the PPARgamma agonists 15-deoxy-Delta(12,14)-PGJ(2) (15d-PGJ(2)) and troglitazone, but not PPARalpha agonist WY-14643, inhibited TNFalpha-induced production of eotaxin and monocyte chemotactic protein-1 (MCP-1) but not IL-8. Eotaxin inhibition was transcriptional and additively enhanced by the glucocorticoid fluticasone and the beta(2)-agonist salmeterol, whereas MCP-1 inhibition was post-transcriptional and additively and synergistically enhanced by fluticasone and salmeterol, respectively. Coimmunoprecipitation revealed that 15d-PGJ(2) induced a protein protein interaction between PPARgamma and the glucocorticoid receptor (GR) in TNFalpha-treated HASM cells, which was enhanced by fluticasone and salmeterol. 15d-PGJ(2), fluticasone, and salmeterol all inhibited TNFalpha-induced histone H4 acetylation at the eotaxin promoter and NF-kappaB p65 binding to the eotaxin promoter and induced PPARgamma and GR association with the eotaxin promoter, as analyzed by chromatin immunoprecipitation assay. Our data suggest that chemokine expression in HASM cells is differentially regulated by PPARgamma agonists and that the interaction between PPARgamma and GR may be responsible for the additive and synergistic inhibition of chemokine expression by PPARgamma agonists, glucocorticoids, and beta(2)-agonists, particularly the chromatin-dependent suppression of eotaxin gene transcription. The interaction may have wide applications and may provide a potential target for pharmacological and molecular intervention. PMID- 15531762 TI - Circulating FGF-23 is regulated by 1alpha,25-dihydroxyvitamin D3 and phosphorus in vivo. AB - Fibroblast growth factor-23 (FGF-23), a novel phosphate-regulating factor, was elevated in hypophosphatemic patients with X-linked hypophosphatemic rickets/osteomalacia and also in patients with chronic kidney disease. These observations suggested the pathophysiological importance of FGF-23 on phosphate homeostasis. However, regulation of FGF-23 production is still unclear. We investigated effects of both dietary phosphorus and 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) on circulating FGF-23 in vivo Administration of. 1alpha,25(OH)(2)D(3) dose-dependently increased serum FGF-23 in thyroparathyroidectomized rats without correlating with serum inorganic phosphorus or serum parathyroid hormone. On the other hand, vitamin D receptor null mice had very low serum FGF-23 and did not respond to the 1alpha,25(OH)(2)D(3) administration. These observations suggested 1alpha,25(OH)(2)D(3) directly or indirectly regulates circulating FGF-23. Serum FGF-23 had a strong correlation with serum inorganic phosphorus controlled by dietary phosphorus in 5/6 nephrectomized rats. High phosphate diet elicited a 5 fold increase in serum FGF-23 compared with sham-operated rats, whereas serum FGF 23 did not correlate with serum calcium or serum creatinine in 5/6 nephrectomized rats. Administration of 1alpha,25-dihydroxyvitamin D(3) also elicited a severalfold increase in serum FGF-23 in the uremic rats. Taken together, this shows that both serum phosphorus and 1alpha,25(OH)(2)D(3) regulate circulating FGF-23 independent of each other. Therefore, we proposed there was a feedback loop existing among serum phosphorus, 1alpha,25(OH)(2)D(3), and FGF-23, in which the novel phosphate-regulating bone-kidney axis integrated with the parathyroid hormone-vitamin D(3) axis in regulating phosphate homeostasis. PMID- 15531763 TI - Platelet factor 4 and interleukin-8 CXC chemokine heterodimer formation modulates function at the quaternary structural level. AB - The apparent complexity of biology increases as more biomolecular interactions that mediate function become known. We have used NMR spectroscopy and molecular modeling to provide direct evidence that tetrameric platelet factor-4 (PF4) and dimeric interleukin-8 (IL8), two members of the CXC chemokine family, readily interact by exchanging subunits and forming heterodimers via extension of their antiparallel beta-sheet domains. We further demonstrate using functional assays that PF4/IL8 heterodimerization has a direct and significant consequence on the biological activity of both chemokines. Formation of heterodimers enhances the anti-proliferative effect of PF4 on endothelial cells in culture, as well as the IL8-induced migration of CXCR2 vector-transfected Baf3 cells. These results suggest that CXC chemokine biology, and perhaps cytokine biology in general, may be functionally modulated at the molecular level by formation of heterodimers. This concept, in turn, has implications for designing chemokine/cytokine variants with modified biological properties. PMID- 15531764 TI - Structure and reactivity of human mitochondrial 2,4-dienoyl-CoA reductase: enzyme ligand interactions in a distinctive short-chain reductase active site. AB - Fatty acid catabolism by beta-oxidation mainly occurs in mitochondria and to a lesser degree in peroxisomes. Poly-unsaturated fatty acids are problematic for beta-oxidation, because the enzymes directly involved are unable to process all the different double bond conformations and combinations that occur naturally. In mammals, three accessory proteins circumvent this problem by catalyzing specific isomerization and reduction reactions. Central to this process is the NADPH dependent 2,4-dienoyl-CoA reductase. We present high resolution crystal structures of human mitochondrial 2,4-dienoyl-CoA reductase in binary complex with cofactor, and the ternary complex with NADP(+) and substrate trans-2,trans-4 dienoyl-CoA at 2.1 and 1.75 A resolution, respectively. The enzyme, a homotetramer, is a short-chain dehydrogenase/reductase with a distinctive catalytic center. Close structural similarity between the binary and ternary complexes suggests an absence of large conformational changes during binding and processing of substrate. The site of catalysis is relatively open and placed beside a flexible loop thereby allowing the enzyme to accommodate and process a wide range of fatty acids. Seven single mutants were constructed, by site directed mutagenesis, to investigate the function of selected residues in the active site thought likely to either contribute to the architecture of the active site or to catalysis. The mutant proteins were overexpressed, purified to homogeneity, and then characterized. The structural and kinetic data are consistent and support a mechanism that derives one reducing equivalent from the cofactor, and one from solvent. Key to the acquisition of a solvent-derived proton is the orientation of substrate and stabilization of a dienolate intermediate by Tyr-199, Asn-148, and the oxidized nicotinamide. PMID- 15531765 TI - Long-term healthcare and cost outcomes of disease management in a large, randomized, community-based population with heart failure. AB - BACKGROUND: Because of the prevalence and expense of congestive heart failure (CHF), significant efforts have been made to develop disease management (DM) programs that will improve clinical and financial outcomes. The effectiveness of such programs in a large, heterogeneous population of CHF patients remains unknown. METHODS AND RESULTS: We randomized 1069 patients (aged 70.9+/-10.3 years) with systolic (ejection fraction 35+/-9%) or echocardiographically confirmed diastolic heart failure (HF) to assess telephonic DM over an 18-month period. Data were collected at baseline and at 6-month intervals. Survival analysis was performed by Kaplan-Meier and Cox regression methods. Healthcare utilization was defined after extensive record review, with an attempt to account for all inpatient and outpatient visits, medications, and diagnostic tests. We obtained data on 92% of the patients, from nearly 53,000 health-related encounters. Total cost per patient was defined by adding estimated costs for the observed encounters, excluding the cost of the DM. Kaplan-Meier analysis showed that DM patients had a reduced mortality rate (P=0.037), with DM patients surviving an average of 76 days longer than controls. Subgroup analysis showed that DM had beneficial outcomes in patients with systolic HF (hazard ratio 0.62; P=0.040), which was more pronounced in NYHA classes III and IV. Although improvements in NYHA class were more likely with DM (P<0.001), 6-minute walk data from 217 patients in whom data were available at each visit showed no significant benefit from DM (P=0.08). Total and CHF-related healthcare utilization, including medications, office or emergency department visits, procedures, or hospitalizations, was not decreased by DM. Repeated-measures ANOVA for cost by group showed no significant differences, even in the higher NYHA class groups. CONCLUSIONS: Participation in DM resulted in a significant survival benefit, most notably in symptomatic systolic HF patients. Although DM was associated with improved NYHA class, 6-minute walk test results did not improve. Healthcare utilization was not reduced by DM, and it conferred no cost savings. DM in HF results in improved life expectancy but does not improve objective measures of functional capacity and does not reduce cost. PMID- 15531766 TI - Randomized clinical trial of abciximab in diabetic patients undergoing elective percutaneous coronary interventions after treatment with a high loading dose of clopidogrel. AB - BACKGROUND: Diabetic patients are at increased risk of adverse outcomes after percutaneous coronary interventions. Although subset analyses suggest particular benefit from the administration of abciximab in diabetic patients, no dedicated large randomized trials have been performed in diabetic patients undergoing percutaneous coronary intervention, and certainly not after pretreatment with a high loading dose of clopidogrel. METHODS AND RESULTS: This study (Intracoronary Stenting and Antithrombotic Regimen: Is Abciximab a Superior Way to Eliminate Elevated Thrombotic Risk in Diabetics [ISAR-SWEET] Study) enrolled 701 diabetic patients with coronary artery disease who underwent an elective percutaneous coronary intervention after pretreatment with a 600-mg dose of clopidogrel >2 hours before the procedure: 351 patients were randomly assigned to abciximab and 350 patients to placebo. The primary end point of the trial was the composite incidence of death and myocardial infarction at 1 year. The frequency of angiographic restenosis (diameter stenosis > or =50%) was the secondary end point. The incidence of death or myocardial infarction was 8.3% in the abciximab group and 8.6% in the placebo group (P=0.91), with a relative risk of 0.97 (95% CI, 0.58 to 1.62). The incidence of angiographic restenosis was 28.9% in the abciximab group and 37.8% in the placebo group (P=0.01), with a relative risk of 0.76 (95% CI, 0.62 to 0.94). The incidence of target lesion revascularization was 23.2% in the abciximab group and 30.4% in the placebo group (P=0.03). CONCLUSIONS: The findings of this study do not support a significant impact of abciximab on the risk of death and myocardial infarction in diabetic patients undergoing percutaneous coronary interventions after pretreatment with a 600-mg loading dose of clopidogrel at least 2 hours before the procedure. The present findings suggest, however, that abciximab reduces the risk of restenosis in diabetic patients receiving coronary bare metal stents. PMID- 15531767 TI - Angiotensin-converting-enzyme inhibition in stable coronary artery disease. AB - BACKGROUND: Angiotensin-converting-enzyme (ACE) inhibitors are effective in reducing the risk of heart failure, myocardial infarction, and death from cardiovascular causes in patients with left ventricular systolic dysfunction or heart failure. ACE inhibitors have also been shown to reduce atherosclerotic complications in patients who have vascular disease without heart failure. METHODS: In the Prevention of Events with Angiotensin Converting Enzyme Inhibition (PEACE) Trial, we tested the hypothesis that patients with stable coronary artery disease and normal or slightly reduced left ventricular function derive therapeutic benefit from the addition of ACE inhibitors to modern conventional therapy. The trial was a double-blind, placebo-controlled study in which 8290 patients were randomly assigned to receive either trandolapril at a target dose of 4 mg per day (4158 patients) or matching placebo (4132 patients). RESULTS: The mean (+/-SD) age of the patients was 64+/-8 years, the mean blood pressure 133+/-17/78+/-10 mm Hg, and the mean left ventricular ejection fraction 58+/-9 percent. The patients received intensive treatment, with 72 percent having previously undergone coronary revascularization and 70 percent receiving lipid lowering drugs. The incidence of the primary end point--death from cardiovascular causes, myocardial infarction, or coronary revascularization--was 21.9 percent in the trandolapril group, as compared with 22.5 percent in the placebo group (hazard ratio in the trandolapril group, 0.96; 95 percent confidence interval, 0.88 to 1.06; P=0.43) over a median follow-up period of 4.8 years. CONCLUSIONS: In patients with stable coronary heart disease and preserved left ventricular function who are receiving "current standard" therapy and in whom the rate of cardiovascular events is lower than in previous trials of ACE inhibitors in patients with vascular disease, there is no evidence that the addition of an ACE inhibitor provides further benefit in terms of death from cardiovascular causes, myocardial infarction, or coronary revascularization. PMID- 15531768 TI - ACE inhibitors for patients with vascular disease without left ventricular dysfunction--may they rest in PEACE? PMID- 15531770 TI - Presentation of cytosolic antigens via MHC class II molecules. AB - Major histocompatibility (MHC) class II molecules function to present antigenic peptides to CD4 T lymphocytes. The pathways by which these molecules present exogenous antigens have been extensively studied. However by contrast, far less is known about the processing and trafficking of cytosolic antigens, which can also serve as an alternative source of ligands for MHC class II molecules. Self proteins, tumor antigens, as well as viral proteins found within the cytosol of cells, can be presented via MHC class II molecules, resulting in the activation of specific CD4 T cells. Studies have begun to reveal unique steps as well as some similarities in the pathways for cytosolic and exogenous antigen presentation. Recent developments in this area are summarized here. PMID- 15531769 TI - C-reactive protein: an activator of innate immunity and a modulator of adaptive immunity. AB - C-reactive protein (CRP) is an acute-phase serum protein and a member of the pentraxin protein family. Its host defense functions predate the adaptive immune system by millions of years. Our current understanding of CRP interactions with complement and with Fcgamma receptors (FcgammaR) have led to an increased appreciation of the regulatory role of CRP in inflammation and autoimmunity. This review outlines the role of CRP in infection, inflammation, and autoimmune disease. We provide a description of recent studies, which suggest that CRP acts through FcgammaR to reduce inflammation and protect from certain autoimmune diseases. A general description of the proposed function of CRP is provided as a framework for future investigation. PMID- 15531771 TI - Osteoblast responses to bacterial pathogens: a previously unappreciated role for bone-forming cells in host defense and disease progression. AB - Although the primary roles of osteoblasts are to synthesize the components of bone matrix and to regulate the activity of bone resorbing osteoclasts, there is growing realization that osteoblasts have an additional function during bone diseases, such as osteomyelitis. Based on our recent studies, we propose a novel role for osteoblasts during bacterial infections of bone, namely, the initiation and maintenance of inflammatory immune responses. In this article, we describe how these nonleukocytic cells can perceive bacterial pathogens of bone to initiate the production of an array of immune regulatory molecules. This pattern of expression is one that could promote the recruitment of leukocytes to sites of bacterial challenge, initiate antigen-specific activation of infiltrating cells, and facilitate the development of cell-mediated host responses to intracellular pathogens of bone tissue, thereby identifying this cell type as a previously unappreciated component in host responses. PMID- 15531772 TI - Manipulating the type 1 vs type 2 balance in type 1 diabetes. AB - Virus infections cause a strong inflammatory reaction that is dominated by the expression of type 1 cytokines and chemokines. Such an aggressive immune response by the host is necessary to eliminate intracellular pathogens. However, because of this shift in the type 1 vs type 2 balance of the immune response, virus infections are potential candidates for triggering autoimmune diseases, such as type 1 diabetes (T1D), herpes stromal keratitis, or multiple sclerosis (MS). In this review we will focus on the pathogenesis of T1D in a virus-induced transgenic mouse model and discuss possibilities of how an aggressive type 1 dominated immune response can be restrained and autoimmunity be abrogated. PMID- 15531773 TI - T-cell activation in the curious world of the intestinal intraepithelial lymphocyte. AB - In conventional terms, when T cells encounter appropriate stimuli, they are induced to undergo molecular and physical changes that confer upon them a state of activation. Once initiated, activation generally results in a state of full T cell responsiveness in an all-or-none manner. Uniquely, however, the intestinal intraepithelial lymphocytes (IELs) bear features that are decidedly different from those of T cells located throughout other immunological compartments in that they exhibit some but not all properties of activated T cells, yet they can be induced to move further into activation provided appropriate costimulatory signals have been received. IEL costimulatory molecules some of which are constitutively expressed, whereas others are upregulated following T-cell receptor (TCR)/CD3 stimulation appear to hold the key to determining the nature and magnitude of the activational process. A system of activation such as this in the intestine would be expected to have great immunological protective value for the host because it would provide an untrammeled process of T-cell activation at a barrier site where the level of antigen exposure is consistently high. Clearly, however, mechanisms must be in place to insure that the IEL activation process is not inadvertently breached. These and other issues central to the operational workings of the intestinal immune system are elaborated in this article, and a model is presented in which IEL activation can be viewed as a layered, three stage activational process. PMID- 15531774 TI - Therapeutic targeting of IL-4- and IL-13-responsive cells in pulmonary fibrosis. AB - Severe forms of idiopathic interstitial pneumonia (IIP), such as usual interstitial pneumonia (UIP), can be impervious to modern steroid and immunosuppressive treatment regimens, thereby emphasizing the need for novel effective therapies. Understanding the cytokine networks that may affect immune and structural cell activation and, hence, the progression of these fatal fibrotic diseases, has been a focus in our research. In this regard, we have examined the role of interleukin (IL)-4 and IL-13 and their respective receptor subunits in this process. Examination of clinical surgical lung biopsies (SLBs) showed that IIP is characterized by the abnormal, heightened expression of the receptor subunits that bind IL-4 and IL-13. Specifically, IL-4Ralpha and IL 13Ralpha2 (the high-affinity IL-13 receptor subunit) was present in greater abundance in SLBs and fibroblasts from IIP patients compared with normal patients, who exhibited no evidence of pulmonary fibrosis. These clinical findings prompted us to investigate whether the targeting of pulmonary cell types that were highly responsive to IL-4 and IL-13 was a viable therapeutic option in IIP. Using a chimeric protein comprised of human IL-13 and a truncated version of an exotoxin from Pseudomonas (abbreviated IL13-PE), we observed that IL13-PE selectively targeted human pulmonary fibroblasts grown from IIP SLBs, whereas it had a minimal effect on fibroblasts grown from biopsies from normal patients. In murine models characterized by abnormal airway or interstitial fibrotic responses, the intranasal administration of IL13-PE significantly attenuated the fibrotic response through the targeting of IL-4Ralpha- and IL-13Ralpha2 expressing pulmonary cells, including monocytes, macrophages, and pulmonary fibroblasts. Together, these data demonstrate that IL-4 and IL-13 are required for the initiation and maintenance of pulmonary fibrosis, and highlight the importance of further investigation of anti-fibrotic therapeutics that prevent the action of both cytokines during clinical pulmonary fibrosis. PMID- 15531775 TI - Dilated cardiomyopathy in retrovirally infected mice: a novel model for silent viral DCM? AB - Dilated cardiomyopathy (DCM) is a clinically relevant disease that can occur independently or secondary to other diseases such as HIV infection and AIDS. To study this latter process, we used a model in which mice are infected with the LP BM5 murine AIDS (MAIDS) retrovirus. Cardiac function of control and infected mice was determined through the in vivo analysis of left ventricular pressure-volume loops. Furthermore, the role of myocarditis was investigated through immunohistochemistry for T-cell, B-cell, and macrophage cardiac infiltrates and Northern blot analysis for tumor necrosis factor (TNF)-alpha and inducible nitric oxide synthase (iNOS). End-systolic and end-diastolic volumes were significantly increased and ventricular stiffness was significantly decreased in infected mice, consistent with DCM; however, no staining for inflammatory cellular infiltrates or TNF-alpha and iNOS was seen. These data support the conclusion that the LP-BM5 HIV model virus causes DCM in the absence of chronic cardiac inflammation. These findings support MAIDS retroviral infection as a new model of idiopathic DCM in which myo-carditis does not occur. PMID- 15531776 TI - HAART drugs induce mitochondrial damage and intercellular gaps and gp120 causes apoptosis. AB - HIV-1 infection is associated with serious cardiovascular complications, but the roles of HIV-1, viral proteins, and highly active antiretroviral therapy (HAART) drugs are not understood. HAART decreases the overall risk of heart disease but leads to metabolic disturbances and possibly coronary artery disease. We investigated toxicities of HIV-1, HIV-1 glycoprotein 120 (gp120), and HAART drugs for human coronary artery endothelial cells (CAECs), brain microvascular endothelial cells, and neonatal rat ventricular myocytes (NRVMs). HIV-1 and gp120, but not azidothymidine (AZT), induced apoptosis of NRVMs and CAECs. Ethylisothiourea, an inhibitor of nitric oxide synthase, inhibited apoptosis induction by gp120. AZT, HIV-1, and gp120 all damaged mitochondria of cardiomyocytes. HAART drugs, AZT, and indinavir, but not HIV-1, produced intercellular gaps between confluent endothelial cells and decreased transendothelial electrical resistance. In conclusion, HIV-1 and gp120 induce toxicity through induction of cardiomyocyte and endothelial cell apoptosis. HAART drugs disrupt endothelial cell junctions and mitochondria and could cause vascular damage. PMID- 15531777 TI - Protecting against peroxynitrite-mediated cytotoxicity in vascular smooth muscle cells via upregulating endogenous glutathione biosynthesis by 3H-1,2-dithiole-3 thione. AB - Peroxynitrite (ONOO(-)) is critically involved in the pathogenesis of cardiovascular diseases. Reaction with glutathione (GSH) was proposed to be a major detoxification pathway of ONOO(-) in the biological system. This study was undertaken to determine if chemically elevated intracellular GSH affords protection against ONOO(-)-mediated toxicity in vascular cells. Incubation of aortic smooth muscle A10 cells with 3H-1,2-dithiole-3-thione (D3T) led to a concentration- and time-dependent elevation of cellular GSH. Treatment of the cells with D3T also augmented protein and gene expression of gamma glutamylcysteine ligase. To examine the effects of D3T-induced GSH on ONOO(-) mediated toxicity, we pretreated A10 cells with D3T and then exposed them to either authentic ONOO(-) or the ONOO(-) generator, 3-morpholinosydnonimine. We observed that D3T pretreatment of A10 cells resulted in a significant protection against ONOO(-) cytotoxicity. Conversely, depletion of cellular GSH by buthionine sulfoximine (BSO) caused a marked potentiation of ONOO(-) cytotoxicity. To further demonstrate the causal involvement of GSH induction in D3T cytoprotection, we cotreated A10 cells with BSO to abolish D3T-induced GSH elevation. BSO cotreatment was found to greatly reverse the protective effects of D3T on ONOO(-)-elicited cytotoxicity. Taken together, our results demonstrate that upregulating GSH biosynthesis by D3T results in a marked protection against ONOO(-)-induced toxicity in vascular cells. PMID- 15531778 TI - Acute ex vivo effects of dimethylbenz[a]anthracene and heat shock on rat tail artery. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants that are known to increase oxidative stress, proteotoxicity, and cytotoxicity in many cell types, but the acute effects of PAHs on vascular contractility are not known. The 70-kDa heat shock protein (HSP70) is known to protect cells against proteotoxicity and cellular apoptosis. Thus, increased HSP70 may be hypothesized to prevent any negative effects of PAHs on vascular smooth muscle. Heat shock treatment is a method used to increase expression of HSPs. Therefore, the objective of this study was to examine the acute (<24-h) ex vivo effects of a PAH (dimethylbenz[a]anthracene; DMBA), heat shock, or a combination of the two treatments on arterial contractility. The results of the present study suggest that acute ex vivo exposure of rat tail arteries to DMBA caused a transient impairment (at 6 h but not 12 h) in contractile response to norepinephrine (NE), but not to depolarization with excess KCl. The DMBA-induced impairment in NE contraction was not explained by a change in 20-kDa myosin light chain (LC20) phosphorylation. Heat shock alone impaired excess KCl-induced contraction and LC20 phosphorylation. Moreover, heat treatment failed to mitigate the effects of DMBA, suggesting that cytotoxicity was not the mechanism of DMBA effect on NE induced contractility. Therefore, both acute ex vivo PAH exposure and heat shock impaired contractility of isolated rat tail arteries in the current study but in independent, noninteracting manners. PMID- 15531779 TI - Activation of inflammatory mediators and potential role of ah-receptor ligands in foam cell formation. AB - Epidemiological data and in vivo animal experiments have indicated that exposure to the Ah-receptor (AhR) ligand dioxin and other dioxin-like compounds can lead to cardiovascular toxicity and atherosclerosis. Here, we investigated the effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), the most potent AhR ligand, on the differentiation of U937 cells into foam cells, which are considered to be early lesions of atherosclerosis. Our findings show that, like oxidized low-density lipoprotein (oxLDL), TCDD promotes the differentiation of U937 macrophages to atherogenic foam cells, verified by lipid accumulation and extensive formation of blebs on the cell surface, which are characteristics of foam cells. Through screening expression patterns of typical genes involved in atherosclerosis and foam cell generation, we could demonstrate that mRNA levels of cyclooxygenase-2, interleukin 1beta, and tumor necrosis factor-alpha were increased in a time- and dose-dependent manner in U937 macrophages treated with TCDD, like oxLDL, and that these changes accompanied significantly elevated levels of matrix-degrading metalloproteinases (MMP)-1, MMP-3, MMP-12, and MMP-13. Increased levels of MMPs were associated with TCDD-stimulated cell migration of U937 macrophages. These findings clearly indicate that AhR ligands, like TCDD, stimulate differentiation of U937 macrophages into potentially plaque-forming foam cells. PMID- 15531780 TI - The oxygen radical generator pyrogallol impairs cardiomyocyte contractile function via a superoxide and p38 MAP kinase-dependent pathway: protection by anisodamine and tetramethylpyrazine. AB - Oxygen-derived free radicals have been demonstrated to contribute to the pathogenesis of myocardial dysfunction, although the underlying mechanism remains not fully understood. This study was designed to examine the role of the superoxide generator pyrogallol on cardiac contractile function and possible intervention with herbal medicines anisodamine and tetramethylpyrazine (TMP) on pyrogallol-induced cardiac contractile response. Adult rat ventricular myocytes were isolated and stimulated to contract at 0.5 Hz. Mechanical properties were evaluated using an IonOptix system including peak shortening (PS), time-to-PS (TPS), time-to-90% relengthening (TR(90)), and maximal velocity of shortening/relengthening (+/-dL/dt). A 10-min exposure of pyrogallol (0 to 10(-2) M) did not affect cardiac contractile mechanics. However, longer duration of pyrogallol exposure (1, 3, and 6 h) significantly shortened resting cell length, reduced PS and +/-dL/dt, and prolonged TPS and TR90 in time- and concentration dependent manners. The pyrogallol (10(-4) M with 6-h incubation)-induced mechanical defects were prevented by the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580 (1 microM) and superoxide dismutase (SOD, 500 U/mL) with the exception that pyrogallol-induced PS depression was unaffected by SOD. Interestingly, incubation of herbal antioxidants anisodamine (10(-7) M) and TMP (10(-7) M) effectively attenuated the pyrogallol-induced cardiac mechanical defects with the exception of PS unaffected by TMP. Our data demonstrate a direct inhibitory effect of pyrogallol on cardiac contraction, probably in a superoxide- and p38 MAP kinase-dependent manner. The antioxidant medicines anisodamine and TMP may be useful in the treatment of oxygen free radical-induced myocardial dysfunction. PMID- 15531781 TI - Gene expression profiles of mouse aorta and cultured vascular smooth muscle cells differ widely, yet show common responses to dioxin exposure. AB - Exposure to environmental toxicants may play a role in the onset and progression of cardiovascular disease. Many environmental agents, such as dioxin, are risk factors for atherosclerosis because they may exacerbate an underlying disease by altering gene expression patterns. Expression profiling of vascular tissues allows the simultaneous analysis of thousands of genes and may provide predictive information particularly useful in early disease stages. Often, however, in vivo experiments are unfeasible for material or ethical reasons, and data from cultured cells must be used instead, even though it may not be known whether cultured cells and live tissues share common global responses to the same toxicant. In a search for genes responsive to dioxin exposure, we used oligonucleotide microarrays with DNA sequences from 13,433 genes to compare global gene expression profiles of C57BL/6 mice aortas with cultured vascular smooth muscle cells (vSMCs) of the same mice. Aorta segments and vSMCs differed in the expression of more than 4500 genes, many showing expression differences greater than 1000-fold. Integration of microarray data into Gene Ontology Project annotations showed that many of the genes differentially expressed belonged to the same biological process or metabolic pathway. Notwithstanding these results, a subset of 35 genes responded in the same fashion to dioxin exposure in both systems. Genes in this subset encoded phase I and phase II detoxification enzymes, signal transduction kinases and phosphatases, and proteins involved in DNA repair and the cell cycle. We conclude that vSMCS may be useful aorta surrogates to study early gene expression responses to dioxin exposure, provided that analyses focus on this subset of genes. PMID- 15531782 TI - Identification of microbial inhibitory functional groups in corn stover hydrolysate by carbon-13 nuclear magnetic resonance spectroscopy. AB - Dilute-acid biomass hydrolysates contain biomass degradation products that are inhibitory to cell growth and fermentation. Overliming with Ca(OH)2 has been found to be one of the most effective methods for detoxifying the dilute-acid hydrolysate for ethanol production. However, the mechanism of overliming is not well understood. Carbon-13 nuclear magnetic resonance (13C-NMR) spectroscopy was used to elucidate the functional groups involved in the overliming reaction. The 13C-NMR spectra showed that the major functional groups removed during the overliming process were aliphatic and aromatic acids or esters, and other aromatic and aliphatic compounds. Ketone and aldehyde functionalities were not detected in the spectra. This is the first time that 13C-NMR has been used to elucidate the overliming reaction. PMID- 15531783 TI - Immobilization of glucoamylase onto novel porous polymer supports of vinylene carbonate and 2-hydroxyethyl methacrylate. AB - Glucoamylase was immobilized onto novel porous polymer supports. The properties of immobilized glucoamylase and the relationship between the activity of immobilized enzyme and the properties of porous polymer supports were investigated. Compared with the native enzyme, the temperature profile of immobilized glucoamylase was widened, and the optimum pH was also changed. The optimum substrate concentration of immobilized glucoamylase was higher than that of native enzyme. After storage for 23 d, the immobilized glucoamylase still maintained about 84% of its initial activity, whereas the native enzyme only maintained about 58% of the initial activity. Moreover, after using repeatedly seven times, the immobilized enzyme maintained about 85% of its initial activity. Furthermore, the properties of porous polymer supports had an effect on the activity of the immobilized glucoamylase. PMID- 15531784 TI - Capacity of Bacillus thuringiensis S-layer protein displaying polyhistidine peptides on the cell surface. AB - S-layer protein of Bacillus thuringiensis strain CTC was used as the carrier protein to display polyhistidine (poly[6His]) peptides on the cell surface. Poly(6His)n was fused with S-layer protein at two different sites, inserting just downstream of the S-layer protein homologous domain (slh) and replacing the non slh region of S-layer protein, respectively. The two series chimeric proteins were both expressed by crystal negative B. thuringiensis strain 4Q7 and strain 171, respectively, as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The recombinant B. thuringiensis cells gained Ni(2+)- and Cd(2+) binding ability and had a capacity to display up to nine copies of poly(6His). The Cd(2+) adsorption quantity of the recombinant strain with the strongest adsorption ability was twice that of the host strain. PMID- 15531785 TI - Xylanase production by the thermophilic mold Humicola lanuginosa in solid-state fermentation. AB - Among the lignocellulosic substrates tested, wheat bran supported a high xylanase (EC 3.2.1.8) secretion by Humicola lanuginosa in solid-state fermentation (SSF). Enzyme production reached a peak in 72 h followed by a decline thereafter. Enzyme production was very high (7832 U/g of dry moldy bran) when wheat bran was moistened with tap water at a substrate--to--moistening agent ratio of 1:2.5 (w/v) and an inoculum level of 3 x 106 spores/10 g of wheat bran at a water activity (aw) of 0.95. Cultivation of the mold in large enamel trays yielded a xylanase titer comparable with that in flasks. Parametric optimization resulted in a 31% increase in enzyme production in SSF. Xylanase production was approx 23 fold higher in SSF than in submerged fermentation (SmF). A threshold constitutive level of xylanase was secreted by H. lanuginosa in a medium containing glucose as the sole carbon source. The enzyme was induced by xylose and xylan. Enzyme synthesis was repressed beyond 1.0% (w/v) xylose in SmF, whereas it was unaffected up to 3.0% (w/w) in SSF, suggesting a minimization of catabolite repression in SSF. PMID- 15531787 TI - Coupled hydroperoxide lyase and alcohol dehydrogenase for selective synthesis of aldehyde or alcohol. AB - The main objective of this work was to improve the selective synthesis of a volatile compound: aldehyde or alcohol using a coupled-enzyme system. A novel method of synthesis of C6-aldehyde or alcohol was carried out in the presence of hydroperoxide lyase (HPLS) activity coupled to alcohol dehydrogenase (ADH) activity. After cleavage of the initial substrate, hydroperoxy fatty acid catalyzed by HPLS, the second enzyme, ADH, can catalyze the reduction of the aldehyde to the corresponding alcohol, or the oxidation of contaminating alcohol into aldehyde, depending on the cofactor present in the medium (oxidized or reduced form). We succeeded in improving the synthesis of one of the products. When coupling HPLS to NADP, the selectivity of hexanal production from 13 hydroperoxy linoleic acid was improved, and hexanol production was reduced 5 to 10 times after 15 min of reaction at 15 degrees C and pH 7.0. In another experiment, HPLS was coupled to ADH in the presence of NADH. The production of alcohol (hexenols) was then favored especially when using 13-hydroperoxy linolenic acid as substrate at concentrations >15 mM, reaching 95% of the products. Coupling of the enzymatic reactions (cleavage reduction) not only reduced the number of steps but also allowed us to increase the conversion rate of the initial substrate (hydroperoxy fatty acid). Structures of the compounds produced in this work were confirmed using gas chromatography-mass spectroscopy analysis. Each of these products has its own delicately different fresh odor that can be used in various applications. PMID- 15531786 TI - Optimization of medium by orthogonal matrix method for submerged mycelial culture and exopolysaccharide production in Collybia maculata. AB - Optimization of submerged culture conditions for the production of mycelial growth and exopolysaccharides (EPSs) by Collybia maculata was investigated. The optimum temperature and the initial pH for EPS production in a shake-flask culture of C. maculata were found to be 20 degrees C and 5.5, respectively. Among the various medium's constituents examined, glucose, Martone A-1, K(2)HPO(4), and CaCl(2) were the most suitable carbon, nitrogen, and mineral sources for EPS production, respectively. The optimum concentration of the medium's ingredients determined using the orthogonal matrix method was as follows: 30 g/L of glucose, 20 g/L of Martone A-1, 1 g/L of K(2)HPO(4), and 1 g/L of CaCl(2). Under the optimized culture conditions, the maximum concentration of EPSs in a 5-L stirred tank reactor was 2.4 g/L, which was approximately five times higher than that in the basal medium. A comparative fermentation result showed that the EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor despite the lower mycelial growth rate. The specific productivities and the yield coefficients in the airlift reactor were higher than those in the stirred-tank reactor even though the volumetric productivities were higher in the stirred-tank reactor than in the airlift reactor. PMID- 15531788 TI - Effect of expression of manganese superoxide dismutase in baculovirus-infected insect cells. AB - It has previously been demonstrated that baculovirus infection of the Spodoptera frugiperda Sf-9 (Sf-9) and Trichoplusia ni BTI-Tn-5B1-4 (Tn-5B1-4) insect cell lines leads to oxidative stress as measured by protein and membrane lipid oxidation and that this oxidative damage contributes to cell death. As a result of these findings, it was hypothesized that baculovirus infection stimulates superoxide radical (O(2)(.-)) synthesis in the mitochondria and that the resulting O(2)(.-) accumulation overwhelms the cells' antioxidant defenses. We investigated the ability of manganese superoxide dismutase (MnSOD) expression (which reduces O(2)(.-) to H(2)O(2) to overcome the oxidative damage caused by baculovirus infection. It was found that MnSOD expression significantly reduced oxidative damage in baculovirus-infected Tn-5B1-4 cells but had no significant effect on oxidative damage in baculovirus-infected Sf-9 cells. The results are consistent with the hypothesis that O(2)(.-) accumulation in the mitochondria is at least partially responsible for the oxidative damage resulting from the baculovirus infection of insect cells. PMID- 15531789 TI - A new maximum likelihood approach with asymmetric residual distribution for multicomponent mass spectra analysis. AB - This paper proposes a new maximum likelihood approach for the deconvolution of identity and quantity of individual compounds based on the multicomponent mass spectra measured by mass spectrometry (MS). Mixture analysis of multicomponent mass spectra is, typically, based on a linear multicomponent mass spectrum model, where the compounds of the measured spectra to be solved are explicitly stated and assumed to be known. In many cases, however, the measured spectrum may contain unknown compounds that are not explicitly stated in the model and a commonly used least square (LS) solution fails. Moreover, a standard improvement over the LS method in these cases, namely the M-estimation (ME) approach, also suffers from this same problem. Our method overcomes the limitations of the LS and ME methods by modeling the effect of the unknown compound(s) to the residual of the linear model. The experimental results presented show that this new approach can separate more robustly the complex multicomponent mass spectra into their individual constituents compared to the LS and ME methods. PMID- 15531790 TI - Comparison of three types of mass spectrometers for HPLC/MS analysis of perfluoroalkylated substances and fluorotelomer alcohols. AB - The performance of three different types of mass spectrometers (MS) coupled to high performance liquid chromatography (HPLC) was compared for trace analysis of perfluoroalkylated substances (PFAS) and fluorotelomer alcohols (FTOHs). Ion trap MS in the full scan and product ion MS2 mode, time-of-flight (TOF) high resolution MS and quadrupole MS in the selected ion mode as well as triple quadrupole tandem MS were tested. Electrospray ionisation in the negative ion mode [ESI-] was best suited for all instruments and compounds. PFAS could only be separated by a buffered mobile phase, but the presence of buffer suppressed the ionisation of FTOHs. Therefore, two independent chromatographic methods were developed for the two compound classes. Mass spectra and product ion spectra obtained by in-source and collision induced dissociation fragmentation are discussed including ion adduct formation. Product ion yields of PFAS were only in the range of 0.3 to 12%, independent from the applied MS instrument. Ion trap MS2 gave product ion yields of 20 to 62% for FTOHs, whereas only 4.1 to 5.8% were obtained by triple quadrupole tandem MS. Ion trap MS was best suited for qualitative analysis and structure elucidation of branched isomeric structures of PFAS. Providing typical detection limits of 5 ng injected in MS2 mode, it was not sensitive enough for selective trace amount quantification. TOF high resolution MS was the only technique combining high selectivity and excellent sensitivity for PFAS analysis (detection limits of 2 to 10 pg), but lacked the possibility of MS-MS. Triple quadrupole tandem MS was the method of choice for quantification of FTOHs with detection limits in the low pg range. It is also well suited for the determination of PFAS, though its detection limits of 10 to 100 pg in tandem MS mode are about one order of magnitude higher than for TOF MS. PMID- 15531791 TI - Determination of rare earth impurities in high purity europium oxide by inductively coupled plasma-mass spectrometry and evaluation of concentration values for europium oxide standard material. AB - Direct determinations of 13 rare earth elements (La, Ce, Pr, Nd, Sm, Gd, Tb, Dy, Ho, Er, Yb, Lu and Y) in high purity europium oxide by inductively coupled plasma mass spectrometry (ICP-MS) have been reported. The operating parameters of the instrument were optimized and the spectral interferences and the matrix effects were investigated. Using Ga or Rh as the internal standard can eliminate most of the matrix effects. The detection limits (3 sigma, n=10, integral time=1s) are 0.013-0.085 ng mL(-1) and the reproducibility (n=11) is 0.9-3%. The recoveries of spiking samples are 80-108%. Europium oxide standard material was made and its concentration values were evaluated by various techniques from 10 collaborating laboratories. By comparison of the results of ICP-MS with the results of other techniques, we have validated that ICP-MS is an accurate and reliable technique for analysis of ultratrace impurities in high purity rare earth matrix. PMID- 15531792 TI - Electrospray ionization mass spectrometry studies of rhenium(I) bipyridyl complexes. AB - Electrospray ionization mass spectrometry (ESMS) has been employed to study the formation of fragment ions of a series of rhenium(I) bipyridyl complexes [(4,4' di-(COOEt)2-bpy) Re(CO)3XPyPF6], where bpy is 2,2'-bipyridine, Py is pyridine, and X is H, 4-methyl, 3-methyl, 4-hydroxyl, 3-hydroxyl, 4-amino, or 3-amino of the pyridine ligand. The effects of substituents (X) on the stabilities of the complexes have been investigated with the increase of fragmentor voltages. For different X, the stabilities of the complexes increase as X become more electron donating from H to CH3, OH, and NH2. For the same substituent, the p-substituted pyridines have stronger stabilizing effect than the corresponding m-substituted ones. Ligand exchange reaction was found in acetonitrile, where the pyridine ligand has been replaced by the solvent indicated by the formation of [M-PF6- XPy+MeCN]+ in the fragmentation. PMID- 15531793 TI - Electrospray mass spectrometry of a series of mixed nitrido 99gTc- heterocomplexes conjugated with bioactive molecules. AB - Electrospray ionization mass spectrometry (ESI-MS) was successfully employed for the identification of six nitrido technetium mixed ligand complexes with a general formula of [99gTc(N)(O,S-BID)(PNP)], where PNP represents a heterodiphosphine and O,S-BID represents a simple dianionic bidentate ligand (compounds 1-3) or a more sophisticated N-substituted O,S-cysteine framework conjugated with a bio- active molecule (BAM) (compounds 4-6). In spite of similar coordination spheres exhibited by all the complexes investigated, simple co ordination compounds 1-3 displayed collisionally-induced fragmentation processes (MSn) different from those observed in biomolecule-containing compounds 4-6. In the latter, more decomposition channels were observed. This behavior is likely to be associated with some additional intramolecular contacts of the biomolecule (or part of the biomolecule) with pendant group(s) incorporated in the PNP-co-ligand. This view is further supported by the observations arising from both in vitro binding affinity experiments and nuclear magnetic resonance investigations. The presence of cationized forms for all compounds 1-6 and the practical lack of the [2M + Na]+ species for biomolecule-containing compounds 4-6 provided further evidence of a subtly different structural conformation. PMID- 15531794 TI - Identification of some polymeric materials by low-temperature pyrolysis mass spectrometry. AB - The application of low-temperature pyrolysis mass spectrometry to the identification of some commercial and synthetic polymers using a direct insertion probe and electron ionization has been studied. The polymers were analyzed directly in the solid probe of a mass spectrometer within the ion source at 70 eV electron impact. The quadrupole mass spectrometer, equipped with a gas chromatogram software and library of low molecular weight compounds, was used to characterize the polymers by measuring the chemical structure of the repeat units and the end groups, and to examine the thermal degradation pathways. The polymers investigated show different degradation pathways. Investigation of sequences and determination of composition of copolymers were studied. Total ion pyrogram and spectrum subtractions were used to separate and measure spectra of pyrolysis steps at distinctly different temperatures. PMID- 15531795 TI - Determination of phenylalanine isotope ratio enrichment by liquid chromatography/time- of-flight mass spectrometry. AB - The application of time-of-flight mass spectrometry to isotope ratio measurements has been limited by the relatively low dynamic range of the time-to-digital converter detectors available on commercial LC/ToF-MS systems. Here we report the measurement of phenylalanine isotope ratio enrichment by using a new LC/ToF-MS system with wide dynamic range. Underivatized phenylalanine was injected onto a C18 column directly with 0.1% formic acid/acetonitrile as the mobile phase. The optimal instrument parameters for the time-of-flight mass spectrometer were determined by tuning the instrument with a phenylalanine standard. The accuracy of the isotope enrichment measurement was determined by the injection of standard solutions with known isotope ratios ranging from 0.02% to 9.2%. A plot of the results against the theoretical values gave a linear curve with R2 of 0.9999. The coefficient of variation for the isotope ratio measurement was below 2%. The method is simple, rapid, and accurate and presents an attractive alternative to traditional GC/MS applications. PMID- 15531797 TI - Fragmentation study of peptides using Fourier transform ion cyclotron resonance with infrared multiphoton dissociation: experiment and simulation. AB - In this study, the fragmentation of gas-phase protonated Angiotensin II is investigated using electrospray ionization (ESI), Fourier-transform ion cyclotron resonance (FT-ICR), and mass spectrometry (MS) with a laser cleavage infrared multiphoton dissociation (IRMPD) technique. The experimental results show that the spectra peaks for the photoproducts are y2/b6- and y7-type ions, corresponding to the cleavage of His-Pro and Asp-Arg in the parent amino acid sequence. The fragmentation of the peptide under collision-free vacuum conditions is modeled using molecular dynamics simulations (MD). The binding energy for the peptide bonds (C'-N bond) of Angiotensin II is estimated from ab initio calculations. The calculations are directed at predicting experimental measurements of the product ions from the photodissociation of the peptide. The product distributions simulated by the MD dissociation trajectories include predominantly y7/b1 and y2/b6 pair ions. PMID- 15531796 TI - Models of fragmentations induced by electron attachment to protonated peptides. AB - Invoking a number of theoretical levels ranging from HF/STO-3G to CCSD(T)/aug-cc pVQZ, we have made a detailed survey of six potential energy surfaces (NH4+, NH4*, [CH3CONHCH3]H+, [CH3CONHCH3]H*, [HCONHCH2CONH2]H+, [HCONHCH2CONH2]H*). In conjunction with this, ab inito direct dynamics calculations have been conducted, tracing out several hundred reaction trajectories to reveal details of the electron-capture dissociation mechanism. The model calculations suggest the possibility of a bimodal pattern where some of the radicals, formed upon recombination, dissociate almost directly within one picosecond, the remaining radicals being subject to complete energy redistribution with a subsequent dissociation occurring at the microsecond timescale. Both processes give rise to c and z backbone fragments, resulting from cleavage of N-Calpha bonds of the peptide chain. PMID- 15531798 TI - Chiral discrimination on the host-guest-complexation of resorc[4]arenes with quarternary amines. AB - The interaction of inherently chiral resorc[4]arenes with different chiral ammonium ions was measured by ESI-MS. For that purpose one enantiomer of the ammonium guests was labeled with deuterium to distinguish the enantiomers by their mass. We synthesized the ammonium salts by reaction of chiral primary amines with either CH3I or CD3I and analyzed the resulting ammonium iodides by NMR and optical rotation. The complexation experiments were performed by mixing the chiral host with various ratios of the unlabeled guest and its labeled enantiomer. By analysis of the integrals of the host-guest complexes we observed a chiral discrimination effect and a secondary isotope effect as well. PMID- 15531799 TI - Changes of phospholipid composition within the dystrophic muscle by matrix assisted laser desorption/ionization mass spectrometry and mass spectrometry imaging. AB - Duchenne muscular dystrophy (DMD) is a neuromuscular disease linked to the lack of the dystrophin, a submembrane protein, leading to muscle weakness and associated with a defect of the lipid metabolism. A study of the fatty acid composition of glycerophosphatidylcholines by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF-MS) and tandem mass spectrometry (MS/MS) enabled us to characterize a change of the lipid composition of dystrophic cells at the time of the differentiation. This modification has been used as a marker to identify with profiling and imaging MALDI-ToF MS regenerating areas in sections of an mdx mouse leg muscle. It is the first time that such a slight change in fatty acid composition has been observed directly on tissue slices using mass spectrometry. This approach will be useful in monitoring the treatment of muscular regeneration. PMID- 15531800 TI - Electrospray mass spectrometry and tandem mass spectrometry of the natural mixture of cyclic peptides from linseed. AB - The cyclic peptides from linseed are composed exclusively of the hydrophobic amino acids: Phe, Leu, Ile, Val, Met, Pro, and Trp. Because these compounds does not contain functional groups which undergo easily protonation or deprotonation. their ionization in solvents used usually for peptide analysis is not efficient. A rapid and sensitive procedure for detection and structure elucidation of the cyclic peptides based on ionization with Na+ and NH4+ ions. A cationisation of methionine containing peptides with methyl iodide has been also described. The extract of seeds of Linum utitatissimum was analyzed directly by ESI-MS and neutral loss ESI-MS/MS technique. The analysis confirms the presence of cyclolinopeptides reported previously: CLA (c(Pro-Pro-Phe-Phe-Leu-Ile-Ile-Leu Val), and CLB (c(Pro-Pro-Phe-Phe-Val-Ile-Met-Ile-Leu)). Cyclolinopeptides CLC, CLD, CLE, and CLG, which contain methionine oxide, were detected in relatively small quantities. These peptides results likely from the oxidation of their not reported precursors: CLD' (c (Pro- Phe-Phe-Trp-Ile-Met-Leu-Leu)), CLE'(c (Pro-Leu Phe-Ile-Met-Leu-Val-Phe)), CLF (c (Pro-Phe-Phe-Trp- Val-Met-Leu-Met), and CLG (c (Pro-Phe-Phe-Trp-Ile-Met-Leu-Met), present at higher concentrations in the extract protected from atmospheric oxygen. The sequences of the unreported cyclic peptides were proposed on the basis of CID experiments and homology with peptides described by Morita,1,2 and supported by the fragmentation of synthetic analogues of CLA of a known structure. PMID- 15531801 TI - N-methyl-N-trimethylsilyltrifluoroacetamide-promoted synthesis and mass spectrometric characterization of deuterated ephedrines. AB - Synthesis and mass spectrometric characterization of drugs or metabolites labeled by stable isotopes has been of great interest in fields of clinical, forensic and doping control analysis. Deuterated ephedrine and p-hydroxypseudoephedrine were prepared from corresponding amines by a novel procedure utilizing N- methyl-N trimethylsilyltrifluoroacetamide and deuterated iodomethane. The mechanism of methylation was studied by mass spectrometry using phenylethylamine as a model compound, and a rearrangement based on an intermediate six-membered ring structure with a trimethylsilyl-enol-ether is proposed giving rise to a leaving group of trimethyliodosilane and the desired monomethylated product. Deuterated analogues to frequently quantitated ephedrines were readily synthesized with purities >90%, and mass spectra recorded under different ionization and dissociation conditions presented distinct fragmentation processes including eliminations of water and methylamine as well as the generation of a benzyl cation. PMID- 15531802 TI - Electrospray tandem mass spectrometry for structural characterization of aporphine- benzylisoquinoline alkaloids. AB - Electrospray mass spectrometry and tandem mass spectrometry techniques were utilized to elucidate the structures of ten aporphine-benzylisoquinoline alkaloids, consisting of monoether link between aporphine and benzyltetrahydroisoquinoline units, which were isolated and identified previously from a variety of Thalictrum sp. (Ranunculaceae family) based mainly on the UV, IR, CD, NMR, EI-MS, CI-MS, derivatization, and chemical degradation techniques. In this investigation, protonated molecules, [M+H]+ ions, for nine tertiary alkaloids, a molecular ion, [M+'] ion, for a quaternary alkaloid, and very intense doubly- protonated molecules, [M+2H]2+ ions (100% of relative abundance) in Q1 Scan MS spectra, and prominent as well as diagnostic product ions for structural information in the tandem MS/MS spectra were observed for all investigated alkaloids each in nanogram quantities. More than 10 microg quantities of each investigated alkaloid or other isoquinoline and aporphine analogs needed for the CI-MS, EI-MS and FAB-MS analysis from the previous studies. PMID- 15531803 TI - High resolution electrospray and electrospray tandem mass spectra of rotenone and its isoxazoline cycloadducts. AB - An evaluation of the gas-phase ion chemistry of rotenone (1) by electrospray ionisation (ESI) mass spectrometry (MS) and tandem mass spectrometry (MS2) is presented, aiming at providing tools for its determination in natural and biological matrices. The behaviour of its cycloadducts with benzonitrile-N-oxide (2) and 2,4,6-trimethylbenzonitrile-N-oxide (3) was also evaluated and the MS data thus obtained have provided evidence into the mechanism of formation of the key product ion at m/z 192 which can be considered a marker in the MS and MS2 spectra of rotenone and its derivatives. PMID- 15531804 TI - Characterization of novel nucleoside analogs by electrospray ionization mass spectra. AB - In this paper, we synthesized a novel nucleoside analog by coupling thymine with dimethyl dicarboxylate biphenyl (DDB). The structure of the target compound was determined using 1H nuclear magnetic resonance (NMR) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). The fragmentation pathways were studied in details through ESI-MS/MS. By comparing with unsubstituted nucleosides, such as AZT, MCI, d4T and DDI, it was found that the nucleoside analog coupled with DDB would not yield the daughter ions corresponding to the fragments of the nucleoside base and arabinofuranose analogs, but would lose a neutral molecule HF and DDB easily. However, the unsubstituted nucleosides could lightly yield the fragment ions of the nucleoside base and sugar ring. Hence, electrospray ionization mass spectrometry combined with tandem mass spectrometry (MS/MS) provides a convenient method to recognize the substituted and unsubstituted nucleosides. PMID- 15531805 TI - Characterization of G-rich and T-rich oligonucleotides using ion-pair reversed phase high-performance liquid chromatography/tandem electrospray ionization mass spectrometry. AB - Characteristics of G-rich and T-rich oligonucleotides were investigated to compare their retention time, total ion current (TIC) intensity, charge-state distribution and product ion using ion-pair reversed-phase high- performance liquid chromatography/tandem electrospray ionization mass spectrometry (IP-RP HPLC/ESI-MS) at room temperature. Three commonly used mobile phases for the analysis of oligonucleotides, triethylammonium acetate (TEAA), triethylammonium bicarbonate (TEAB) and triethylammonium hexafluoroisopropanol (HFIP) have been utilized. Retention time of G-rich and T-rich oligonucleotides was significantly different in TEAA and TEAB buffer systems, while in the HFIP buffer system it was affected more by the length of oligonucleotides. On the other hand, the ESI-MS ion abundance in the HFIP buffer system was higher than that in both TEAA and TEAB buffers. The TIC intensity of T-rich oligonucleotides was much higher than that of G-rich oligonucleotides in all mobile phases. In addition, much higher charge-state fragments were observed in HFIP buffer system than that in the case of TEAA and TEAB buffer systems. Product ions of both G-rich and T-rich oligonucleotides were affected by charge state of parent ions and collision energy. PMID- 15531806 TI - Characterization of the O-4 phosphorylated and O-5 substituted Kdo reducing end group and sequencing of the core oligosaccharide of Aeromonas salmonicida ssp salmonicida lipopolysaccharide using tandem mass spectrometry. AB - The molecular structure of the wild strain of the lipopolysaccharide core of Aeromonas salmonicida, ssp salmonicida has been sequenced using tandem mass spectrometry. The core oligosaccharide was determined to contain an O-4 phosphorylated and O-5 substituted Kdo reducing group, and its structure is proposed as the follows: [structure: see text] After the core oligosaccharide of LPS was released from the lipid A portion by conventional treatment with 1% acetic acid, we demonstrated the existence of a homogeneous mixture composed mainly of the native core oligosaccharide containing the Kdo with its O-4 phosphate group intact, and a degraded core oligosaccharide mixture, which eliminated the O-4 phosphate group with extreme facility. The precise molecular structure and glycone sequence of the homogeneous mixture of phosphorylated and dephosphorylated core oligosaccharides was determined by electrospray ionization (ESI) mass spectrometry and tandem mass spectrometric analysis. CID-MS/MS of the homogeneous mixture of permethylated core oligosaccharides afforded a series of diagnostic product ions which confirmed the established sequence of the glycones to be determined. Matrix-assisted laser desorption/ionization (MALDI) tandem mass spectrometry reconfirmed the molecular structure of the dephosphorylated homogeneous permethylated mixture of the core oligosaccharides containing the diastereomeric 4,8- and 4,7-anhydro-alpha-keto acids. PMID- 15531807 TI - Mass spectrometric identification of cyclic polysulfides in sediment of the Eastern Gulf of Finland. I. AB - Structures of six cyclic polysulfides, previously unknown as organic environmental pollutants, were analyzed from a sediment sample from the Eastern Gulf of Finland. The determinations were done by gas chromatography connected to mass spectrometry. High resolution (HRMS) measurements of the isotopic composition of four compounds could be done to confirm their molecular formulae. Total low resolution (LRMS) spectra were used to elucidate structures of all six compounds by thermochemical approach, application of fragmentation rules and by ICLU simulation of the spectra. The compounds were deduced to be (in the order of GC- retention) 1,2,4-trithiacycloheptane, tetrathiacyclopentane, 1,2,4,5 tetrathia-cyclohexane, 1,2,3,4- tetrathiacycloheptane, 1,2,3,4 tetrathiacyclohexane and 1,2,4,6-tetrathiacyclooctane. PMID- 15531808 TI - Mass spectrometric identification of cyclic polysulfides in sediment of the Eastern Gulf of Finland. II. AB - Nine polysulfides, previously unknown as environmental organic pollutants, were analyzed from a sediment sample from the Eastern Gulf of Finland. The determinations were done by gas chromatography connected to low- and high resolution mass spectrometers. The structure of the polysulfides was elucidated by determination of isotopic composition of abundant molecular and fragment ions by high-resolution mass spectrometry (HRMS), the interpretation of ion structures in low-resolution mass spectra (LMRS) using the thermochemical approach, the application of fragmentation rules and performing the ICLU simulation of abundance of ions in isotope clusters. Seven compounds were known in literature, but mass spectra were reported for only two of them, both recorded from mixture. Struture evaluation was successful for other substances, but not for the substance(s) in first eluting GC peak, where HRMS date were not obtained. Suggested structures of the cyclic compounds were (in the order of GC retention) 3,4-dithiacyclohexene, 1,2,3-trithiacyclohexane, 3,4,5- trithiacyclohexene, 1,2,4 trithiacyclohexane, cyclopropyl hydrodisulfide, 1,2-dithiole-3-thione and 1,2,3,4,5-penta- thiacyclo-octane. One acyclic congener identified was dimethyl tetrasulfide. PMID- 15531809 TI - ACU Column 15.4 (November 2004). Connected separateness or separate connection: envisioning body with mind. PMID- 15531810 TI - The changing landscape for the elimination of racial/ethnic health status disparities. AB - The elimination of racial/ethnic health status disparities is a compelling national health objective. It was etched in sharp relief by the 1985 report of the U.S. Department of Health and Human Services Secretary's Task Force on Black and Minority Health and considerable attention has been devoted to the problem since that report. But the problem persists, disparities are not fully explained and effective policies to reduce them have been elusive, a situation presenting both opportunities and challenges. Important advances towards reducing racial/ethnic health disparities may be made by better understanding the complex bidirectional relationship between and among the multiple factors, biological and non-biological, influencing morbidity and mortality. The landscape in which these influences are felt is anything but static. In this paper selected components of the landscape that are critical to the elimination of racial/ethnic health status disparities are reviewed. These factors underscore the importance of adopting and maintaining a perspective on health disparities that encompasses a broad array of health determinants. PMID- 15531811 TI - Pediatric emergency department outreach: solving medical problems or revealing community solutions? AB - Low-acuity (LA) use of emergency departments (EDs) is often viewed as misuse or abuse. We designed a program to help users access services more efficiently. Community health workers (CHWs) functioned as health educators, screeners, and liaisons to care. A participatory curriculum emphasized medical problems. Qualitative ethnographic methods were used for formative evaluation. Ninety families received regular visits from CHWs. Original system-oriented objectives and methods did not fit community needs. Instead, information was gathered regarding service utilization patterns: (1) families mistrusted primary and preventive care, reporting frequent discrimination and humiliation; (2) a primarily biomedical prioritization did not match families' need hierarchies; (3) complex demands of poverty made other uses of the system challenging; (4) primary care services were frequently avoided; and (5) the ED was often preferred. Intangible psychosocial factors and practical complexities of poverty were powerful promoters of LA visits. Families were creative strategists, viewing LA visits as rational use, not misuse, of a challenging system. PMID- 15531812 TI - Neighborhood poverty, mortality rates, and excess deaths among African Americans: Philadelphia 1999-2001. AB - We used vital statistics and census data to determine whether mortality rates in Philadelphia were associated with neighborhood poverty, and to what extent excess mortality among African Americans was associated with neighborhood poverty. Gender-specific, age-adjusted mortality rates for 1999-2001 were strongly associated with neighborhood poverty among both women and men overall, and among both African Americans and non-Hispanic whites. The actual number of deaths among African Americans was 5,305 higher than it would have been if African Americans had had the same gender- and age-specific mortality rates as the average for non Hispanic whites in Philadelphia, and 1,944 higher than if African Americans had had the same gender- and age-specific rates as non-Hispanic whites in the same neighborhood poverty categories. The excess mortality associated with neighborhood poverty and the socioeconomic factors that force large numbers of African Americans into poverty and high-poverty neighborhoods appear to be major factors in excess mortality among African Americans. PMID- 15531813 TI - Missed opportunities for patient education and social worker consultation at The Arbor Free Clinic. AB - We conducted a cross-sectional survey of 210 patients who came to a free medical clinic for health care over an 8-month period. We (1) measured their satisfaction with care, (2) determined the frequency of missed opportunities for providing health education and social work consultation, and (3) assessed whether patient specific factors drive the frequency of these missed opportunities. Of the 210 patients surveyed, a total of 168 (80.0%) completed the entire survey. The mean satisfaction rating was high (4.6 on a scale of 1 to 5). A significant number of missed opportunities occurred, with only 28% of patients receiving patient education material, and 32% of patients visiting the social worker. No particular patient groups emerged as most susceptible to these missed opportunities. This study shows both the high degree of patient satisfaction at this free clinic and the many opportunities for improving patient education and social services. Adding health education and social work consultation to the patient encounter could improve the health of these patients. PMID- 15531814 TI - The Jane Dent Home: the rise and fall of homes for the aged in low-income communities. AB - The Jane Dent Home was established in 1898 (as the Home for Aged and Infirm Colored People) to serve African American elderly barred from admission to most homes for the aged. Sustained by community leadership through difficult times, the Home finally closed in 1975 after growing and persistent racial and economic segregation of Chicago's low-income neighborhoods combined with pressure from state government to ensure fire safety. This history illustrates the decline of not-for-profit homes for the aged while for-profit nursing homes were capturing market share. In Chicago this trend is strongest in low-income communities of color, which may lead to lower quality of care for such communities. Support for indigenous not-for-profit long-term care may promote the goals of health care equity articulated by Healthy People 2010. PMID- 15531815 TI - Racial and ethnic differences in utilization of health services in patients with diabetes enrolled in medicaid. AB - We evaluated racial and ethnic differences in use of medical care between patients with diabetes enrolled in Medicaid and explored whether differences varied by state Medicaid program. Using data from 137,006 patients we created a multivariable Poisson regression model to examine the effect of race on ambulatory care visits, emergency ward visits, and hospitalization rates for patients with diabetes mellitus enrolled in three state Medicaid programs. We found significant differences in service use between groups, which varied depending on state. For example, black patients compared with whites had significantly fewer outpatient visits but more hospitalizations in New Jersey; by contrast, blacks had higher outpatient visit rates and lower hospitalization rates in Georgia. Racial and ethnic differences in health service use among Medicaid enrollees were not consistent across states, suggesting that local factors, including varied Medicaid policies, may affect racial and ethnic differences in use of health care services. PMID- 15531816 TI - Attitudes and beliefs among Mexican Americans about type 2 diabetes. AB - Hispanics in the United States have a disproportionately high risk for non insulin-dependent diabetes mellitus (type 2 diabetes) compared with non-Hispanic whites. Little is known of the attitudes and beliefs about diabetes in this group. Using data from six focus groups of 42 Mexican Americans (14 men and 28 women), we characterized perceptions about the causes of and treatments for type 2 diabetes. Many participants believed diabetes is caused by having a family history of the disease, eating a diet high in fat or sugar, and engaging in minimal exercise. Experiencing strong emotions such as fright (susto), intense anger (coraje), or sadness and depression (tristeza) was also thought to precipitate diabetes. Nearly all participants expressed the belief that it is important to follow doctors' recommendations for diet and exercise, oral medication or insulin; many also cited herbal therapies, such as prickly pear cactus (nopal) and aloe vera (savila) as effective treatments. These findings may be useful in designing interventions to reduce the burden of diabetes in Hispanic populations. PMID- 15531817 TI - Building an academic-community partnership for increasing representation of minorities in the health professions. AB - We evaluated collaboration among academic and community partners in a program to recruit African American youth into the health professions. Six institutions of higher education, an urban school system, two community organizations, and two private enterprises became partners to create a health career pipeline for this population. The pipeline consisted of 14 subprograms designed to enrich academic science curricula, stimulate the interest of students in health careers, and facilitate entry into professional schools and other graduate-level educational programs. Subprogram directors completed questionnaires regarding a sense of common mission/vision and coordination/collaboration three times during the 3 year project. The partners strongly shared a common mission and vision throughout the duration of the program, although there was some weakening in the last phase. Subprogram directors initially viewed coordination/collaboration as weak, but by midway through the project period viewed it as stronger. Feared loss of autonomy was foremost among several factors that threatened collaboration among the partners. Collaboration was improved largely through a process of building trust among the partners. PMID- 15531818 TI - Characteristics of patients at three free clinics. AB - This study examined factors associated with the use of three free clinics located in Central Massachusetts. A total of 248 patients completed a questionnaire during the 2-month study period. Descriptive results showed a majority of free clinic patients are low-income, uninsured, and female. Many patients (62%) do not have a usual source of care, nor do they know where to go if the clinic is not open (61%). Most (82%) report using free clinics because they lack insurance. Patients who had been using the free clinics longer than 1 year are more likely to use the clinic because of inadequate insurance (p = 0.002) and as a way to obtain prescription drugs (p < 0.001). Although they serve an important need, free clinics cannot provide comprehensive, continuous care. Efforts to provide health care to the medically underserved must take these findings into consideration if they are to be successful. PMID- 15531819 TI - Socioeconomic differences in health: how much do health behaviors and health insurance coverage account for? AB - As evidence accumulates that both unhealthy behaviors and inadequate access to health care are responsible in part for poor health, there is a tendency to attribute the differences in health status between the poor and the affluent to the higher prevalence of unhealthy behaviors and inadequate access to health care among people of low socioeconomic status (SES). The purpose of this study is to determine quantitatively how much health behaviors and health insurance coverage account for the SES disparity in health. The study employed secondary analysis of data collected through the Kentucky Behavioral Risk Factor Surveillance System for 2000. After adjusting for health behaviors and health insurance coverage, the differences in health among different levels of SES (measured by education and income) remained strong and significant. Health behaviors and health insurance coverage accounted for 10-16% of the socioeconomic differences in health. PMID- 15531820 TI - How do urban African Americans and Latinos view the influence of diet on hypertension? AB - Uncontrolled hypertension and its complications continue to be major health problems that disproportionately affect poor minority communities. Although dietary modification is an effective treatment for hypertension, it is not clear how hypertensive minority patients view diet as part of their treatment, and what barriers affect their abilities to eat healthy diets. We conducted nine focus groups with 88 African American and Latino patients treated for hypertension to assess their knowledge, attitudes, behaviors, and beliefs concerning hypertension. Participants generally agreed that certain foods and food additives play an important role in the cause and treatment of hypertension. However, they found clinician-recommended diets difficult to follow in the context of their family lives, social situations, and cultures. These diets were often considered expensive, an unwelcome departure from traditional and preferred diets, socially isolating, and not effective enough to obviate the need for medications. These findings suggest the importance of culturally sensitive approaches to dietary improvements. PMID- 15531821 TI - Self-reported health of residents of the Mississippi Delta. AB - The rural Lower Mississippi Delta of Arkansas, Louisiana, and Mississippi has a large economically and socially disadvantaged population at high risk for health problems. Their health status is poorly understood as they are not well represented in national health surveys. A random-digit-dialing telephone survey was conducted in 2000, with 2,236 respondents representing residents of 36 counties along the Mississippi River. Self-reported chronic conditions, health status, and obesity (derived from weight and height) were compared with the nationally representative Continuing Survey of Food Intake of Individuals. High cholesterol, diabetes, and hypertension were significantly higher than in the national sample. Obesity was strikingly higher in Delta children (27.9% versus 16.2%) of all ages and in Delta adults (33.9% versus 17.3%). Controlling for age, income, and gender, African Americans were at particular risk for obesity, hypertension, and diabetes. A public health crisis appears to exist in the Delta given the high prevalence health problems. PMID- 15531822 TI - Provision of on-site medical care to patients with hepatitis C in drug treatment units. AB - Substance abusers are at high risk for hepatitis C (HCV) infection and also constitute a group that is medically underserved and hard to reach. We conducted a nationwide survey with 445 randomly selected drug treatment units in the United States to determine unit and patient characteristics associated with the provision of on-site medical services for HCV-infected drug users. Eighty-four percent of the 322 units that estimated having at least one HCV-infected patient reported that they provided patients with HCV-related medical care. Drug treatment units were more likely to provide at least some of this care on site if they were residential, part of a network, or affiliated with a hospital; had medical staff; and required that their patients undergo a medical examination before entering treatment. Some organizational factors appear to influence the provision of on-site medical services to HCV-positive patients in drug treatment units. Further research on the role of such factors could inform the development of effective models of care for patients with hepatitis C in drug treatment organizations. PMID- 15531823 TI - Medicaid coverage and utilization of adult dental services. AB - This study evaluated the economic impact of a policy change in adult Maryland Medicaid dental benefits that eliminated reimbursements to dentists. We examined all claims for 2 years before and after the change. Reimbursements to dentists fell to zero from their preenactment period annual rate of US 7.6 million dollars; other care settings simultaneously generated an additional US 232,470 dollars savings during the postenactment period. Medicaid's goal to reduce costs was achieved; however, disadvantaged patients may have been confused by the policy change and likely suffered poorer health outcomes and paid for treatment out of pocket, found free clinics, or received free care from generous dentists. PMID- 15531824 TI - Are women with self-reported cognitive limitations at risk for underutilization of mammography? AB - Functional limitations (namely, limitations in activities of daily living and instrumental activities of daily living) have previously been demonstrated to exert a negative influence on mammography utilization. This study examines self reported cognitive limitation in addition to sociodemographic, functional, and other health-related factors to determine their relationship with self-reported mammography use in the previous year. Data from the 1998 National Health Interview Survey was analyzed for 6,053 women, ages 50 years and older. Just over 44% of women with self-reported cognitive impairment (n = 351) reported a mammogram in the previous year, compared with 55% of unimpaired women (n = 5,702). Logistic regression analysis indicates that the presence of a cognitive limitation significantly reduced the likelihood of a mammography in the previous year (p < 0.05) after controlling for other sociodemographic, functional, and health-related factors. Women with self-reported cognitive limitations were 30% less likely than unimpaired women to utilize mammography after controlling for various forms of disability and other factors. Thus, women with cognitive impairments may be at risk for underutilization of mammography and therefore at risk for later-stage breast cancer diagnoses. PMID- 15531827 TI - Diarrhea prevention through food safety education. AB - OBJECTIVE: Food safety education (FSE) was imparted in order to reduce the prevalence of diarrhea in children and improving the knowledge, attitude and practices of mothers regarding safe feeding practices. METHODS: The study was conducted on mothers (n=200) of underprivileged children (6-24 months) through Anganwadi workers (AWW). The food safety education package incorporated three messages: washing hands with soap and water, avoid feeding leftover food and keep surrounding clean. These were imparted using lectures, slogans, posters, charts, flash cards, role-play. Calendars and leaflets were also handed over to the mothers as education material. After food safety education intervention, the households were studied again as at the baseline, on various aspects. RESULT: There was 52% reduction in the incidence of diarrhea. The environmental sanitation and personal hygiene scores of most of the households and mothers improved. Improvement in the practice of hand-washing and avoidance of feeding leftovers to children was seen. There was reduction in the microbial load in the hand rinse samples of both mothers and children. CONCLUSION: AWW proved to be effective change agents and FSE was successful in bringing about a positive behavior modification in mothers. PMID- 15531828 TI - Nosocomial diarrhea. AB - OBJECTIVE: To study the occurrence of nosocomial diarrhea in pediatric wards and the role of infections in its causation. METHODS: 75 cases conforming to the CDC definition of nosocomial diarrhea were selected for the study. Stool samples were subjected to direct microscopic examination, bacteriological culture and direct antigen detection for Clostridium difficile (C. difficile) and Rota virus. RESULTS: All the cases were sporadic in origin. Rota virus was detected in 16.67% cases and Enteropathogenic Escherichia coli (E.P.E.C.) was detected in 12% cases. Cases in which no pathogen was detected (82.7%) outnumbered those in which a pathogen was identified (17.3%). CONCLUSION: Rota virus and E.P.E.C were the most common etiological agents. C. difficile was detected in one case only. Most of the sporadic cases of nosocomial diarrhea in pediatric patients can be managed on fluids and electrolytes. PMID- 15531830 TI - Pulmonary function tests in Indian girls--prediction equations. AB - OBJECTIVE: The present study was conducted to evaluate pulmonary functions and develop prediction equations in Indian girls. METHODS: 1038 normal healthy schoolgirls in the age group of 5 to 15 years were selected for the present study. The anthropometrical parameters like height, weight, sitting height and chest circumference were measured and body surface area (BSA) and fat free mass (FFM) were derived using equations. The pulmonary functions such as FEV1, FVC, FEV1/FVC% and PEFR were studied. RESULTS: The height, sitting height, weight, BSA, chest circumference, body fat as well as FEV1, FVC, FEV1/FVC% and PEFR were comparable with Indian standards. Multiple regression equations were developed to predict FEV1, FVC and PEFR using anthropometrical indices like height, fat free mass and age or chest circumference in view of significantly high correlation of these parameters with lung function variables [height and FEV1 (r-0.90), height and FVC (r-0.899), height and PEFR (r-0.891), chest and FEV1 (r-0.868), chest and FVC (r-0.867), chest and PEFR (r-0.83)]. CONCLUSION: The regression equations to predict the pulmonary functions were presented using the independent variables like height, fat free mass and chest circumference or age, since these variables have shown very strong predictability for FEV1, FVC and PEFR. The equations presented in this study can be considered as referral standards for Indian girls. PMID- 15531829 TI - Retinopathy of prematurity--risk factors. AB - OBJECTIVE: Improved survival of low birth weight, premature babies have increased the incidence of retinopathy of prematurity. This hospital-based, prospective, study was undertaken to determine its incidence and risk factors in our neonatal unit. METHODS: Neonates with gestational age < or = 35 weeks and/or birth weight < or = 1500 gm born over a one-year period were examined by indirect ophthalmoscopy between 2 to 4 weeks after birth, and followed up till retinal vascularisation was complete. Maternal and neonatal risk factors were noted and data analyzed by statistical package SPSS-10.0. RESULTS: Sixty babies were thus examined. The incidence of retinopathy was 21.7% in the cohort, 33.3% in babies < or = 32 weeks gestation and 36.4% in babies weighing < or = 1250 gm. Oxygen (p=0.01), sepsis (p=0.04) and apnoea (p=0.02) were independent risk factors. Retinopathy was significantly more severe in babies with hyaline membrane disease (p=0.02) and lower birth weight (p=0.02). Severe disease was never seen before 6.5 weeks of age. CONCLUSION: Indirect ophthalmoscopy should be performed at 4 weeks of post natal age in all preterm babies with birth weight < or = 1500 gm, and intensified in the presence of risk factors like oxygen administration, apnoea and septicemia. PMID- 15531831 TI - Safety of blind liver biopsy. AB - OBJECTIVE: The aim of the study was to observe the safety of blind liver biopsy in children. METHODS: One hundred fifteen consecutive liver biopsies in hospitalized children were evaluated retrospectively. Using a standard percussion technique biopsy sites were chosen and through intercostal space blind liver biopsies were performed by TruCut biopsy needle. RESULTS: The first biopsy sample was considered macroscopically adequate in 94.8% of cases. A definitive histological diagnosis was possible in 99.1% of cases. Sixty nine children were more than 5 years of age and of these 8 (11.6%) complained of pain at the biopsy site. External hemorrhage from the biopsy site was seen in 1 (0.9%) case but no sign of internal hemorrhage was detected during the 24 hours follow up period. No child died following the procedure. CONCLUSION: Blind liver biopsy in the studied hospitalized children was found to be a safe procedure. PMID- 15531832 TI - Infantile colic--unfolded. AB - Infantile colic is a common complaint for which mothers bring their babies to pediatricians. Though the condition has been a cause of controversy and a subject of extensive research, no consensus is yet reached on its definition, etio pathogenesis and management. This article reviews the existing literature on present subject. PMID- 15531833 TI - Diapers and skin care: merits and demerits. AB - Diapers have been used for care of babies since decades to prevent soiling and for social convenience. However, the use of diaper poses a risk of developing diaper dermatitis. Recent innovations in diaper technology have led to development of superabsobent disposable diapers, emollient delivering diapers and breathable diapers. These newer types of diapers reduce the incidence of diaper dermatitis. The non-biodegradable material used in superabsorbent diapers is, however, a matter of serious concern because of its toxic effects and environmental pollution. The different type of diapers, recent advances in them and their environmental concerns are discussed in this article. PMID- 15531834 TI - Management of meningococcemia. AB - Meningococcemia causes substantial morbidity and mortality worldwide, usually in the term of fulminant disease. This paper reviews the background, pathophysiology, clinical manifestations, and treatment of this entity, along with prevention measures and public health issues to be considered. The authors present updated information on breakthroughs in the understanding of genetic predisposition to invasive disease. The status of adjunctive treatment modalities such as monoclonal antibodies and activated protein C, and progress in conjugate vaccine development and implementation are also addressed. PMID- 15531835 TI - Group A beta-hemolytic streptococcal bacteremia. AB - OBJECTIVE: The aim of this study was to review the clinical features, laboratory findings and the risk factors associated with invasive group A streptococcal infections in children admitted to our institution over a 9-year period (January 1, 1990 through December 31, 1999). METHODS: Medical records of children who had a positive blood culture for group A beta-hemolytic streptococci and children who had this organism isolated from any other sterile site were identified and retrospectively reviewed. RESULTS: Forty-one children with invasive GAS were identified, of whom 15 (36%) were diagnosed between 1990 and 1994, while the balance (26 patients, 63%) were diagnosed between 1995 and 1999 (p< 0.05). The mean age was 4.3 +/- 2.5 years (age range: 2 months to 16 years). Thirteen (32%) patients were infants. Sixteen patients had only bacteremia, while 25 patients had in addition to bacteremia the following: cellulitis (n: 13), osteomyelitis (n: 6), pneumonia (n: 3), meningitis (n: 1), pharyngitis (n:3) and Toxic Shock Syndrome (n: 2). Primary varicella infection constituted the most common predisposing factor for invasive GAS infections and occurred in 11 (27%) patients. Leukocytosis (A white blood cell count > 15,000/mm3) occurred in 21 (51 %) patients, while leukopenia (A white blood cell count < 5000/mm3) occurred in 2 patients. Parenteral crystalline penicillin G followed by oral penicillin or amoxicillin were the most common antibiotics administered. The mean hospital length of stay was 8 days (range: 6 -32 days). All, but one patient survived. The one patient who died had malnutrition and died from streptococcal toxic shock syndrome. CONCLUSION: More cases of invasive GAS were diagnosed during the second half of the study period, however, the overall rate of occurrence of bacteremia during the study period was consistent with previous reports. Primary varicella infection was the most common predisposing factor for invasive GAS infections. The low occurrence of toxic shock syndrome and fatalities among children with invasive GAS infections are consistent with other pediatric but not with adult series. PMID- 15531836 TI - Management of neonatal herpes simplex virus infections. AB - As many as 2,500 infants develop neonatal herpes each year, most of whom are born to women with no history or physical findings suggestive of genital herpes. Infection usually takes one of three forms: 1) disease localized to skin, eyes, and mucous membranes, 2) localized central nervous system infection, or 3) disseminated infection. Exposure to the virus occurs during passage through an infected birth canal, but 5% of infants acquire the infection in utero. The mortality rate is 31% for disseminated infection and 6% for localized central nervous system disease; long-term neurologic sequelae are seen in 17% and 70% of survivors, respectively. Diagnosis is made by isolating of the virus from skin lesions or other involved sites. The polymerase chain reaction for the detection of viral DNA in cerebrospinal fluid or serum is now the diagnostic test of choice for central nervous system or disseminated neonatal herpes because it has higher sensitivity than traditional culture methods. Treatment is with high-dose intravenous acyclovir (60 mg/kg per day in three divided doses), with adjustments made for infants with renal or hepatic insufficiency. Supportive measures and neuroimaging studies are often required. Acyclovir is administered for three weeks, but infants with disease localized to the skin, eyes, and mucous membranes can be treated for two weeks if the cerebrospinal fluid polymerase chain reaction assay is negative for herpes simplex virus DNA. Prevention of infection in infants can be accomplished by cesarean delivery when women have active lesions at the onset of labor. Neonates delivered through an infected birth canal should be screened between 24 and 48 hours of age with viral cultures of eyes, nasopharynx, mouth, and rectum. If positive, they should be treated with acyclovir even if asymptomatic. Suppressive acyclovir therapy beginning at 36 weeks gestation is often prescribed for women with frequent recurrences of genital herpes. PMID- 15531837 TI - Clinico-hematological profile of congenital leukemia. AB - Congenital leukemia is a rare disorder that usually presents with extramedullary infiltrates and myeloid phenotype in 80% cases. The authors present 4 cases of congenital leukemia diagnosed over 11 year period, three of which were of acute lymphoid leukemia and one was of biphenotypic leukemia. PMID- 15531838 TI - Tyrosinemia type I: a clinico-laboratory case report. AB - Progressive hepatocellular dysfunction in a neonate, resulting in elevated serum alpha-fetoprotein together with raised blood levels of tyrosine and methionine, a generalized amino aciduria and the absence of urinary delta-aminolevulinic acid and succinylacetone, suggests a diagnosis of tyrosinemia type Ib. Classical tyrosinemia type I arises from a deficiency of fumarylacetoacetate hydrolase while the variant tyrosinemia type Ib results from a deficiency of maleylacetoacetate isomerase. PMID- 15531839 TI - Maroteaux-Lamy syndrome. AB - Mucopolysaccharidoses are a type of lysosomal storage disorders characterized by defect in the degradation of Mucopolysaccharides due to deficiency of specific lysosomal enzymes leading to their accumulation in various tissues. MPS -VI (Maroteaux-Lamy Syndrome) is an autosomal recessive syndrome due to deficiency of enzyme Aryl- Sulfatase -B, and is characterized by characteristic facies, normal intelligence, Dysostosis multiplex, organomegaly, joint stiffness, corneal clouding and striking inclusions in peripheral blood leucocytes. We present an 8 year-old male child with MPS-VI syndrome, confirmed by enzyme assay. PMID- 15531840 TI - Wilson's disease--unusual features. AB - Two cases of Wilson's disease with unusual features are reported. In one case neurological abnormality was the presenting feature without any clinical involvement of the liver. In the other case, neurologic manifestations were associated with rickets and cholelithiasis, a result of chronic hemolytic state. Apart from clinical profile both the cases were diagnosed by grossly reduced serum ceruloplasmin level. However, Kayser-Fleischer rings were found in each case. PMID- 15531841 TI - Cytodiagnosis of yolk sac tumor. AB - To analyze cytomorphologic features of yolk sac tumors of childhood. Four cases of pediatric yolk sac tumor (YST), diagnosed by fine needle aspiration cytology were reviewed (1998-2002). Age of patients ranged from 1(1/2) to 5 years. Three cases presented clinically with an intra-abdominal mass while one case presented with a testicular mass. Fine needle aspirates had been obtained directly as well as under radiologic (USG/CT) guidance. Smears were stained with H & E and Papanicolaou stain. In all cases values of serum alpha-fetoprotein and hCG were available preoperatively. Histopathologic diagnosis was correlated with cytologic findings in all the cases. Cytologic examination showed richly cellular smears with a combination of morphological patterns. Characteristically, tumor cells were arranged in papillary groups, tight cell clusters and formed acinar structures. Cells showed enlarged, moderately pleomorphic, hyperchromatic nuclei and moderate amount of cytoplasm, some of which displayed cytoplasmic vacuolation, displacing the nuclei eccentrically. Preoperatively, serum alpha fetoprotein level was raised in all cases. Histopathology confirmed the cytologic diagnosis. Yolk sac tumor is common among the germ cell tumors of pediatric age group which presents a spectrum of cytomorphologic features having important differences with other germ cell neoplasm, e.g. embryonal carcinoma. Clinicoradiologic features and tumor markers are additionally helpful for an accurate cytologic diagnosis. PMID- 15531842 TI - Dyke-Davidoff-Masson syndrome (DDMS). PMID- 15531843 TI - Renal agenesis in extrahepatic portal vein obstruction. PMID- 15531844 TI - Prevalence of anemia amongst pregnant mothers and children in Delhi. PMID- 15531845 TI - Hypomelanosis of ito. AB - Four-year-old boy presented with hypopigmented skin lesions since birth and seizure of recent onset. He had psychomotor retardation and hemi hypertrophy of left half of body. Computerised tomography scan findings were suggestive of hemimegalencephaly. On clinical grounds, a diagnosis of Hypomelanosis of Ito was made. PMID- 15531846 TI - Incomplete monosymptomatic leptomeningeal angiomatosis. AB - Phakomatoses or neurocutaneous syndromes are an important cause of seizures in the pediatric age group. The Sturge-Weber syndrome may affect the eye, skin and brain at different times. The skin lesions need not always manifest. We report a case of isolated affection of the central nervous system in a case of Sturge Weber syndrome in the absence of ocular or cutaneous manifestations. Our case qualifies to be called incomplete monosymptomatic leptomeningeal angiomatosis. PMID- 15531847 TI - Diabetes mellitus in an infant. AB - Onset of diabetes in the first year of life is uncommon, most cases being those of transient diabetes of the newborn. We report the case of a child who presented at 10 weeks of age with diabetic keto-acidosis, and continues to be insulin dependent at 18 months. PMID- 15531848 TI - Hemihypertrophy and primary small intestinal lymphangiectasia in incontinentia pigmenti achromians. AB - A rare occurrence of primary small intestinal lymphangiectasia in an 8-year-old girl with incontinentia pigmenti achromians is reported. In addition, she had right sided hemihypertrophy. Though intestinal lymphangiectasia is known to have a few syndromic associations, its co-existence in a child with incontinentia pigmenti achromians has not yet been reported in the literature. Hemihypertrophy is also extremely rare in the latter and only very few instances of an association between the two have been documented previously. PMID- 15531849 TI - Glutaric aciduria type I associated with learning disability. AB - The authors report a 7-year-8-months-old boy with glutaric aciduria type I who had associated dyslexia, dysgraphia and dyscalculia. The diagnosis of glutaric aciduria type I was confirmed on the basis of characteristic neuroimaging and biochemical findings. Axial T1-weighted magnetic resonance imaging scan of the brain showed fronto-temporal atrophy, open opercula and bat-wing dilatation of the sylvian fissures. Axial T[2]-weighted and FLAIR imaging showed hyperintense signal abnormality in both putamen and in the fronto-parietal deep white matter. Urinary aminoacidogram by thin layer chromatography revealed a generalized aminoaciduria. Urinary organic acid analysis by gas chromatography- mass spectroscopy revealed a marked excretion of glutaric acid. Psychoeducational testing was used to diagnose the learning disability. We postulate that the accumulation of glutaric acid and other metabolites was responsible for the child developing the associated learning disability. PMID- 15531850 TI - Gastric volvulus with perforation in association with congenital diaphragmatic hernia. AB - Gastric volvulus in children is a rare surgical emergency. We describe 2 cases of gastric volvulus with gastric perforation in association with congenital diaphragmatic hernia. One case presented in infancy after recurrent chest infections with intestinal obstruction. The other case presented in the early neonatal period. Both cases underwent laparotomy with repair of the stomach and diaphragm defect. The first case developed pyoperitoneum postoperatively, was reexplored but succumbed to septicaemia. The second baby had a favourable postoperative outcome. PMID- 15531851 TI - A dancer with an unusual cause of neck pain. AB - A girl presented with a dull ache in the neck and mild difficulty in neck movements. She had limited clinical signs and her initial work up failed to reveal the cause. With the help of imaging modalities and CT guided needle biopsy, she was diagnosed to have an eosinophilic granuloma of the fifth cervical vertebra. There were no neurological symptoms. She was successfully managed with immobilization of spine, local irradiation and systemic vinblastine. The literature is briefly reviewed for clinical features, diagnosis and management. PMID- 15531852 TI - Bilateral congenital split hand with tibial aplasia. AB - The authors report on a child with bilateral typical split-hand (ectrodactyly) and tibial aplasia, probably the first Indian report. Minor limb malformations in the maternal side suggests an autosomal dominant inheritance. The clinical spectrum and the inheritance of this malformation are discussed. Prenatal screening in families with such malformations is essential. PMID- 15531853 TI - Acephalus acardiac fetus. AB - Acephalus Acardia is an uncommon congenital malformation due to twin reverse arterial perfusion syndrome. In this condition the donor twin provides circulation for itself and the recipient acardiac twin. The acardiac twin is grossly abnormal with severe reduction anomalies of the upper part of the body. We report a case with autopsy findings and review of literature. PMID- 15531854 TI - [Recurrent spontaneous miscarriages and hyperhomocysteinemia]. AB - AIM: Recurrent fetal loss is defined as the number of consecutive miscarriages which is not less than 2 occurred within the 16th week of gestation and it is a very interesting pathology of pregnancy. Further to thrombophilia, very important causes have been identified, since the damage of the vascular system supporting the placenta may cause a deficiency of placenta functions and development, leading to a loss of the conception product, also in a condition of hyperhomocystinemia, causing a damage to the vascular endothelium. Hyperhomocystinemia seems to be a risk factor for artero-venous thrombotic diseases, even not in pregnancy. METHODS: We have examined 40 patients referred to our Institute for unexplained fetal loss (at least 2 consecutive miscarriages within the 16th week of gestation) and the same number of patients who had at least 1 spontaneous delivery with a healthy and alive newborn and none abortion nor fetal death nor abruptio placentae. RESULTS: The mean levels of homocystinemia observed were significantly different in the 2 groups (p = or < 0.05). In the control group the values of plasmatic homocysteine were 10+/-4 micromol/L, corresponding to normal range, while in the other group the values of plasmatic homocysteine were 21+/-6 micromol/L, values certainly elevated, also because during the 1st trimester of pregnancy the levels of homocysteine decrease, reaching the lowest value during the 2nd trimester of pregnancy. In particular, high levels of homocysteine have been found in 25% (10) of women with unexplained early fetal loss. CONCLUSION: Hypercystinemia, as a consequence of an interaction between a primary genetic defect and a nutrition condition (folate deficiency), may be a cause of recurrent miscarriages. Therefore, in these cases, a possible hypercystinemia should be searched and an association of folic acid and vitamin B6, a non teratogenic treatment, should be useful to restore the metabolic picture and to favour the pregnancy outcome. PMID- 15531855 TI - [Screening for pre-eclampsia in a low-risk population at 24 weeks: uterine artery Doppler flow velocimetry and genetic variants of factor V, prothrombin and methylenetetrahydrofolate reductase]. AB - AIM: Pre-eclampsia is one of the major causes of maternal and fetal morbidity and mortality. The aim of this study was to evaluate the clinical usefulness of screening of genetic thrombophilic mutations and uterine artery Doppler flow velocimetry at 24 weeks of gestation in the prediction of pre-eclampsia in low risk pregnant women. METHODS: We performed the genetic analysis for Leiden mutation of factor V gene (FV), G20210A mutation of the prothrombin gene (PT) and C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene in 103 women that had already attended routine ultrasonography scanner at 24 weeks at our Department. RESULTS: The frequency of heterozygous carriers of the Leiden FV was 17.4% in women with pre-eclampsia and abnormal artery Doppler flow velocimetry compared with 3.12% in patients with normal pregnancies. This difference was statistically significant (P<0.05). The frequency of mutation G20210A of PT gene was 1.5% vs 4.3% between women with normal pregnancies and with pre-eclampsia. This difference is not statistically significant. The frequency of homozygous patients for the C677T mutation of MTHFR gene among patients with pre-eclampsia was 21.7% and in the control group was 10.3%, but this difference is not statistically significant. No thrombophilic genes variants were found in women with pre-eclampsia and normal uterine artery Doppler flow velocimetry. CONCLUSION: We demonstrated the important association between FV Leiden mutation, abnormal uterine artery Doppler flow velocimetry at 24 weeks and pre-eclampsia in our low-risk population. PMID- 15531856 TI - [Human papillomavirus of the female genital tract: prevalence in women attending a centre for early cancer detection]. AB - AIM: The authors study the trend of cytologic human papillomavirus (HPV) detection in the period between 1980 and 2000 in women attending a clinic for genital cancer early detection. METHODS: Routine activity, besides pap smear, is based on clinical evaluation, colposcopy and target biopsy in the presence of a lesion. The cytological trend is compared with that of histologic and clinical findings of HPV lesions on the basis of data prospectively collected. RESULTS: About 5% of cytologic HPV detection is observed in the whole, with a pick of about 11% in the years 1992-1994, and values around 2-3% up to 1991 and after 1995. The finding does not change if the whole pap smear group is considered (46,862) or only the first pap smear per woman (16,810). A similar trend, but with a smaller pick, is observed in histologic findings, not in colposcopic findings. The clinical finding of condylomata is higher in the first period of observation, sloping down through the time intervals considered. Standardization on age or patient's recruiting does not change the evidence. CONCLUSION: The trend of the HPV parameters studied could represent an epidemiological model for HPV infection. The clinical epidemics is followed by a period of HPV cytological pick, which might be followed, at suitable time interval, by more cervical intraepithelial neoplasia/squamous intraepithelial lesions, according to the well known latency between infection and premalignant changes. PMID- 15531857 TI - Chlamydia trachomatis prevalence in North-West Italy. AB - AIM: Chlamydia (C.) trachomatis infection is the most common sexually transmitted disease (STD) among sexually active adolescents and young adults in Europe. The aim of this study was to determine the prevalence of C. trachomatis among sexually active women in Piedmont, Italy and the correlation between some risk factors and C. trachomatis infection. METHODS: In our study 31,419 sexually active women aged 12-55 were screened for C. trachomatis by Abbott's ligase chain reaction (LCR) using cervical swabs during the period 1997-2001 at St. Anna Obstetric-Gynecological Hospital, Turin. All the patients answered a specific questionnaire. RESULTS: In our analysis the prevalence of C. trachomatis infection was found to be 1.23%, and the average age among the infected patients was 36.98 years. Statistical analysis was performed using the chi squared test; a p<0.05 was considered significant. A correlation was found between a positive test result and the age at the first intercourse (p<0.001), the number of sexual partners in the preceding 6 months (p<0.001), the presence of symptoms (p<0.001), a low level of education (p<0.001) and an East-European and Central-Northern African citizenship (p<0.001). No statistically significant differences were found among the contraceptive methods used, whether an hormonal or a barrier type; a marked increment of the risk was observed when no contraception was used. CONCLUSION: Frequent microbiological examinations are desirable for patients whose anamnesis shows an augmented risk of contracting sexually transmitted infections in order to avoid long term complications from misdiagnosed or asymptomatic pathologies, as often happens with C. trachomatis. PMID- 15531858 TI - Effects of soy isoflavones on menopausal neurovegetative symptoms. AB - AIM: Dietary soy supplementation may reduce the incidence of hot flushes in postmenopausal women due to the presence of phytoestrogens in soy foods. The aim of this study was to assess the effects of a 3-month administration of different doses of either oral or transdermal soy isoflavones on climacteric complaints in postmenopausal women. METHODS: Sixty postmenopausal women were assigned randomly to one of the following groups: groups A and B received 50 mg/day or 75 mg/day of oral isoflavones respectively, while groups C and D received 6 mg/day or 12 mg/day of transdermal isoflavones. A control group (group E) was composed of 15 patients that did not receive any treatment. Hot flushes, Kupper-man index and vaginal dryness at baseline and then after 5, 9 and 13 weeks were evaluated. RESULTS: The isoflavone treatment led to a progressive significant (p<0.01) reduction of the number of hot flushes in the groups A, B and D with a dose related effect in group D. At week 13, the Kupperman index and hot flush score decreased significantly in all the treated groups, while vaginal dryness score was reduced only in group B. CONCLUSION: Our findings suggest that both oral and transdermal soy isoflavones are effective to attenuate slight to moderate menopausal neurovegetative symptoms with a dose-related effect. PMID- 15531859 TI - [Dinoprostone: slow release vaginal insert (Propess) and intracervical gel (Prepidil) for the induction of labour with unriped cervix]. AB - AIM: The purpose of the present study is to compare the effectiveness and safety of a slow release vaginal PGE2 insert (Propess) with intracervical PGE2 gel (Prepidil gel) in the induction of cervical ripening and labour. METHODS: For the induction of labour we selected 103 single pregnancies at term presenting a Bishop score of less than 5. Fifty-one were induced with Propess, and 52 with intracervical Prepidil. RESULTS: The 2 groups were homogeneous as regards indications to induction and obstetric characteristics. The success of induction (achievement of uncomplicated vaginal delivery) was comparable in the 2 groups: Propess 67%, Prepidil 65%. The times needed to induce labour were on average longer with Propess (16 h 59 min) than with Prepidil (12 h 54 min), (p<0.05); nevertheless the time needed to achieve delivery by the vaginal route within 24 hours was comparable (49% vs 48%). The number of patients requiring more than one application of prostaglandin was less in the Propess group (5.9%) than in the Prepidil group (55.8%) (p<0.001). The times relative to dilation and expulsion did not differ significantly. Resort to cesarean section for fetal indication (cardiotocographic changes) was greater in inductions with Prepidil (8 cases) compared to Propess (2 cases), p<0.05. CONCLUSION: The systems proved equally effective, nevertheless Propess seems to be safer thanks to the lower incidence of cardiotocographic changes such as to indicate urgent cesarean section. Propess would seem to be more acceptable on the part of patients thanks to the smaller number of applications necessary. PMID- 15531860 TI - Pathophysiology, diagnosis and treatment of endometriosis. AB - Endometriosis is generally a benign condition. However, there is a tendency towards progression and in part deep infiltrating processes occur, which can lead to obstruction and even silent loss of organ function such as kidney failure. Hereditary, genetic and immunological peculiarities have been found. Important are symptoms such as pain and infertility, which burden the life of the women involved. In spite of extensive research in the past the pathogenesis and epidemiology of the disease are not definitely clarified as yet. The estimated prevalence of endometriosis ranges between 10% and even more than 50% depending on the underlying problems of the women studied. Besides the main location of the disease within the pelvis also extra pelvic locations have been found (lung, lymph nodes, scars). The diagnosis should be established by histological verification. The main therapeutic modalities consist of surgery and medical treatment or combinations, which need to be individually adjusted, because of the high risk of recurrence and progression of the disease. The aim should be early detection and treatment as complete as possible, since symptom relief, resolution or prevention of infertility and decrease of recurrences are at their best under these circumstances. PMID- 15531861 TI - The genetics, diagnosis and treatment of amenorrhea. AB - The menstrual cycle is regulated by complex feedback interactions between the ovaries, pituitary and hypothalamus. A disruption at any point in one of these pathways may lead to irregularities in the menstrual cycle. In particular, amenorrhea, the cessation of menstrual functioning, serves as an indicator of ovarian, pituitary and/or hypothalamic dysfunction. Historically, diagnosing and treating amenorrhea presented medical professionals with numerous practical difficulties. In recent years, however, studies of amenorrhea have yielded new understandings and new treatments of the disorder. This paper synthesizes these current methodologies for diagnosing, treating and understanding both primary and secondary amenorrhea. PMID- 15531862 TI - [Current surgical management in ovarian cancer recurrences]. AB - Ovarian cancer is a tumor with a high trend of recurrence and this occurrence consistently increases the difficulty of the patient cure and reduces the efficacy of current treatments. The role of surgery in persistent or recurrent ovarian cancer is controversial and the type of surgery can be different according to the different stages and invasion of tumor; it can be a debulking surgery followed by chemotherapy (to eradicate the most part of ovarian cancer, leaving a minimal tumoral residue), an interval surgery (for advanced ovarian cancer stage in previously operated patients, followed by 2 or 3 inductive chemotherapy cycles and subsequently a cytoreductive redo surgery) and a cytoreductive secondary surgery, after optimal primary surgical treatment and minimal tumoral recurrence. In some cases it is possible either to perform a debulking surgery during a primary (after the conclusion of primary treatment) or a salvage or palliative surgery (to improve, after an acceptable time period, clinical symptoms in patients with progressive cancer or resistant to treatments). The aims of surgical therapy, to be performed in a patient with ovarian cancer relapse, are to reduce, as much as possible, the tumour size, to increase the quality of life and to increase the survival time; in this review different surgical techniques to be carried out in each case, selected for disease staging, for tumour cells kinetic and for surgical goals, are discussed. PMID- 15531863 TI - [Male contraception]. AB - The only safe method of male contraception is vasectomy, with high reversibility secured by microsurgery. Italy, however, suffers from a lack of regulations on this subject. Hormonal treatment (testosterone plus progestational hormones) is far from providing reliability and safety, while some perspectives, theoretical only for the time being, are offered by studies on functional infertility induced by either speeding up (ganglioplegic, sympathomimetic, parasympatholytic, oxytocin, endothelin, angiotensin) or inhibiting (sympatholytic) the sperm transport through the epididymis, or altering the epididymal environment (alpha chloridin, chlorodeoxyglucose). PMID- 15531864 TI - [Spontaneous uterine rupture in the third trimester of gestation after laparoscopic myomectomy. A case report]. AB - Uterine rupture in pregnancy in women who undergone previous laparoscopic myomectomy is an exceptional event. The spread of laparoscopy in the surgical treatment of uterine myomas, however, has been accompanied by the occurrence of cases of uterine rupture in pregnancy. The case we report is the sixth present in the literature. It regards a 34-years-old woman who suffered uterine rupture at the 26th week of gestation. PMID- 15531865 TI - [Therapy and prevention of obstetric hemorrhage by tamponade using a balloon catheter]. AB - The aim of this study was to verify the possibility to use a balloon catheter as hemostatic method in pregnancies at high risk for hemorrhage as an alternative to less conservative surgical procedures. In 4 pregnancies at risk for hemorrhage, a compressive endouterine or endocervical method, consisting of a balloon catheter filled of warm saline solution, was used in order to stop or to prevent otherwise uncontrollable bleeding. In the cases treated the use of the balloon catheter was safe and effective both in the treatment of hemorrhage due to abnormal placentation and in the prevention of hemorrhagic complications due to abnormal implantation of pregnancy. Among novel medical and surgical approaches developed to control obstetric hemorrhage, tamponade using a balloon catheter could be considered a valid option. Moreover, this procedure, by avoiding radical surgical treatments, such as hysterectomy, which are frequently performed in unstable patients, could offer the advantage of preserving fertility. PMID- 15531866 TI - [Ovulation induction with gonadotropin in patients with thalassemia pretreated with pulsatile GnRH: outcome]. PMID- 15531867 TI - [Cytomegalovirus infection in pregnancy]. PMID- 15531868 TI - Maternal complications from tocolytic treatment with ritodrine. Three cases of pulmonary edema. PMID- 15531869 TI - Acardius acephalus as one of triplets. PMID- 15531870 TI - A 24-month survey on root canal treatment performed by NiTi engine driven files and warm gutta-percha filling associated system. AB - AIM: Techniques based on NiTi engine driven files for shaping and warm gutta percha for obturing the root canal space are becoming more and more popular. Aim of this paper is to evaluate, by a clinical longitudinal study, the type of sealing and the outcome of endodontic treatment, performed by new Profile and Thermafil technique, in teeth with or without periapical lesions. METHODS: A total of 122 teeth in 64 patients were consecutively enrolled in the study; 63 teeth had normal periapical status (Group A) and 59 teeth had periapical lesion (Group B). Endodontic therapy was carried out by Profile .04 and Profile .06 NiTi engine driven files and the obturation was made by Thermafil, a filling technique based on heated gutta-percha surrounding a plastic carrier. The type of sealing was evaluated by a score (A best - D worst) and the type of healing was classified as complete, incomplete and failure according to previous published data. The radiographical outcome of the teeth was evaluated by independent observers after 24 months. Data were analysed by non parametric statistics. RESULTS: At the end of the study, 115 teeth (59 group A and 56 group B) were examined. The quality of sealing was evaluated and some differences were reported comparing different types of teeth. The radiographic outcome was evaluated and 94.9% showed to be completely healed in group A and 48.2% in group B (p=0.0001). CONCLUSION: The technique based on NiTi engine driven files and Thermafil heated gutta-percha has shown, after 24 months, results which were comparable to other root canal shaping and filling methods. PMID- 15531871 TI - Long term follow-up of patients affected by temporomandibular joint dysfunction treated with orthodontic and/or prosthetic occlusal therapy. AB - AIM: Clinical evidences about therapeutic effectiveness of splint therapy and modern concept concerning the etiopathogenesis of temporomandibular disorders (TMD) lead to a review of the therapeutic approach accepted until the middle 90s for the solution of this derangement of the stomathognatic system. Aim of this study is to produce a long term follow-up of a sample of TMD patients, to estimate the validity and utility of the present therapeutic procedures. METHODS: After diagnostical classification of the patients observed, they have been included in a therapeutic protocol. The outcomes obtained (46 patients at the 1st control in 1994 and 38 patients at the last control in 2001) are re-examined with an average follow-up of 10 years. RESULTS: The results obtained showed the efficacy of the therapeutic procedures carried out (36 healed, 9 improved and 1 worsened at the 1994 check-up: on the whole, a successful result in 98% of cases; 17 healed, 21 improved and 8 lost at the 2001 check-up: on the whole, a successful result in 100% of cases), but at present their application has to be reviewed according to a more conservative and careful therapeutic approach, based on the evaluation of some fundamental parameters. CONCLUSION: The rehabilitative occlusal therapy, in its various aspects, is successful in the long term treatment of TDMs, as long as it is used only when it is possible to determine a correct indication. PMID- 15531872 TI - Secondary bone grafting of alveolar cleft. AB - AIM: The study evaluates the repair of residual alveolar cleft through secondary bone graft, consisting in the transplantation of autologous bone to restore the continuity of the maxillary arch and achieve normal functioning and esthetics. METHODS: During 2001-2002, 15 patients (age range 9-26 years; 7 males, 8 females) were submitted to secondary bone graft at the Maxillo-facial Surgery Operative Unit, University Hospital, Sassari. Eleven patients had complete unilateral cleft, 4 had complete bilateral cleft. All patients were operated upon by the same surgeon; they received a graft of autologous bone from the iliac crest. For preoperative and postoperative evaluation at 1 year, the following were utilised: plaster casts of the tooth arches, OPT, photographs and complete clinical documentation. RESULTS: Postoperative results were: 100% formation of a bone bridge between the maxillary segments; 70% closure of oro-nasal fistula; 100% maxillary stability; 80% spontaneous eruption of the canine within the graft; 70% height of alveolar ridge level I, 25% level II, 5% level III; 70% orthodontic closure; 80% optimal periodontal condition and 20% presence of gingival recession. In 1 subject the graft site became infected, in 4 cases an oro-nasal fistula remained. CONCLUSION: This method was found to be the most valid one at present. The best period to intervene is during late childhood (9 years). Results and functional and esthetic recovery were satisfactory and encouraging to continue utilising this technique. PMID- 15531873 TI - Therapeutic objectives and surgical treatment of mandibular condyle fractures. Personal experience. AB - AIM: A review of condylar fractures treated ''non-surgically'' and ''surgically'' over the last 5 years (1996-2001) at the Maxillofacial Surgery Division of the Brescia Civil Hospital (Italy) is presented. The non surgical and surgical treatments carried out, as well as their effectiveness and therapeutic aims, are illustrated. METHODS: From June 1996 to June 2001, at the Maxillofacial Surgery Division of the Brescia Civil Hospital, 179 fractures of the mandibular condyle were treated. Of the 179 cases treated, a distinction was immediately made between ''non-surgical'' (103 cases) and ''surgical'' cases (76 cases). The 76 ''surgical'' cases were all treated with the same surgical technique. Out-patient follow-up was at days 20, 60, 90, and 1 year, together with a radiographic check up at 60 days. Of the 76 condylar fractures treated surgically, 15 patients selected at random were re-examined; the medical history was investigated and clinical and radiological examinations performed. For each patient, the type of condylar fracture and whether this was associated with other mandibular fractures were recorded. The following data were also recorded: age of the patient; pain at the surgical site or at the mandible at the time of check-up; presence of normal occlusion and laterodeviation; any surgical sequelae at the facial nerve on the operated side; maximum mouth opening; any radiological defect; time since surgery. Then, the patient was asked to give his/her own evaluation of the surgical outcome, by expressing adequate, good or excellent result. RESULTS: In patients treated surgically the indication is almost absolute in the following cases: condylar fractures types II and IV of the Spiessl/Schroll classification, low or high condylar fractures with medial dislocation of the condyle; non surgical treatment cannot be applied for reasons of patient's health; condylar fracture associated with other open fractures of the face. The use of rigid fixation also may help in the success of the technique adopted. In all the cases (surgical and non-surgical) it is important to achieve early mobilization of the mandible and carry out a rehabilitation program for masticatory function. CONCLUSION: The goals of condylar treatment may therefore be summarized as follows: to achieve a good occlusion; to eliminate pain; to achieve a proper mandibular symmetry; to achieve good mandibular movement in the 3 planes of space. PMID- 15531874 TI - Observation of titanium pretreatment effects using a scanning electron microscope and study of titanium/ceramic and titanium/resin adhesive interface. AB - AIM: A morphological analysis is made of the titanium surface sandblasted using different sized microparticles, as well as an analysis of the adhesive interface after having layered the ceramic and resin on the same prepared surface. METHODS: A total of 14 titanium samples were prepared and randomly divided into 4 groups according to their different preparation methods. Group 1: control, no surface treatment. Group 2: 50 microm aluminium dioxide treatment. Group 3: 150 microm aluminium dioxide treatment. Group 4: Rocatec Plus system treatment. Samples of groups 2, 3, and 4 were subdivided into 2 groups: A) only sandblasted, B) sandblasted and ceramic (2B and 3B) or resin (4B) layered on top. The treated surface and the interface were then observed by scanning electrom microscope. RESULTS: Group 2 surface appeared irregular and rough; the interface with ceramic appeared discontinuous. Group 3 surface presented larger and deeper cavities where ceramic penetrated completely. Group 4 samples presented a rough surface covered by silicate particles and the interface between titanium and resin appeared regular and continuous. CONCLUSION: In order to guarantee an adequate interface, the 150 microm pretreated titanium surface seemed more suitable than the 50 microm one. The Rocatec Plus system was successful in obtaining an optimal layering with the resin due to its excellent quality. PMID- 15531875 TI - Implant-prosthetic rehabilitation after excision of hard palate gland carcinoma. AB - In Summer 2002, a patient with frequent epistaxis was admitted to the San Gerardo Hospital. Local examination showed a big mass in the nasal fossa. TC and RMN revealed a diffuse erosion of the palatal bone and infiltration of the maxilla. The tumor was removed and histological examination showed a gland carcinoma. The young age of the patient and the need of an adequate bone reconstruction led to use osteoperiosteal calf bone graft. Subsequently, using local anesthesia, 6 implants were positioned. After normal healing and the period of osteointegration, a Toronto bridge was made on implants that the patient wears with no problem. PMID- 15531876 TI - Home care maintenance protocol for ear prostheses. AB - Hygienic home care maintenance for patients who have received a maxillofacial prosthesis is very important in order to secure a long-term success of a facial rehabilitation. Craniofacial implant therapy requires a specific protocol to care for the peri-implant tissues and material used for the prosthesis. This article presents the maintenance and care protocol for proper follow-up management of a maxillofacial ear prosthesis. Home care methods for the prosthesis hygiene are described, including: the insertion and the disconnection of the ear prosthesis, hygiene of the bar abutments before and after postdefinitive connection to the craniofacial fixtures, washing of the device, professional follow-up. PMID- 15531877 TI - The Microprocessor complex mediates the genesis of microRNAs. AB - MicroRNAs (miRNAs) are a growing family of small non-protein-coding regulatory genes that regulate the expression of homologous target-gene transcripts. They have been implicated in the control of cell death and proliferation in flies, haematopoietic lineage differentiation in mammals, neuronal patterning in nematodes and leaf and flower development in plants. miRNAs are processed by the RNA-mediated interference machinery. Drosha is an RNase III enzyme that was recently implicated in miRNA processing. Here we show that human Drosha is a component of two multi-protein complexes. The larger complex contains multiple classes of RNA-associated proteins including RNA helicases, proteins that bind double-stranded RNA, novel heterogeneous nuclear ribonucleoproteins and the Ewing's sarcoma family of proteins. The smaller complex is composed of Drosha and the double-stranded-RNA-binding protein, DGCR8, the product of a gene deleted in DiGeorge syndrome. In vivo knock-down and in vitro reconstitution studies revealed that both components of this smaller complex, termed Microprocessor, are necessary and sufficient in mediating the genesis of miRNAs from the primary miRNA transcript. PMID- 15531878 TI - Lipocalin 2 mediates an innate immune response to bacterial infection by sequestrating iron. AB - Although iron is required to sustain life, its free concentration and metabolism have to be tightly regulated. This is achieved through a variety of iron-binding proteins including transferrin and ferritin. During infection, bacteria acquire much of their iron from the host by synthesizing siderophores that scavenge iron and transport it into the pathogen. We recently demonstrated that enterochelin, a bacterial catecholate siderophore, binds to the host protein lipocalin 2 (ref. 5). Here, we show that this event is pivotal in the innate immune response to bacterial infection. Upon encountering invading bacteria the Toll-like receptors on immune cells stimulate the transcription, translation and secretion of lipocalin 2; secreted lipocalin 2 then limits bacterial growth by sequestrating the iron-laden siderophore. Our finding represents a new component of the innate immune system and the acute phase response to infection. PMID- 15531879 TI - Processing of primary microRNAs by the Microprocessor complex. AB - Mature microRNAs (miRNAs) are generated via a two-step processing pathway to yield approximately 22-nucleotide small RNAs that regulate gene expression at the post-transcriptional level. Initial cleavage is catalysed by Drosha, a nuclease of the RNase III family, which acts on primary miRNA transcripts (pri-miRNAs) in the nucleus. Here we show that Drosha exists in a multiprotein complex, the Microprocessor, and begin the process of deconstructing that complex into its constituent components. Along with Drosha, the Microprocessor also contains Pasha (partner of Drosha), a double-stranded RNA binding protein. Suppression of Pasha expression in Drosophila cells or Caenorhabditis elegans interferes with pri miRNA processing, leading to an accumulation of pri-miRNAs and a reduction in mature miRNAs. Finally, depletion or mutation of pash-1 in C. elegans causes de repression of a let-7 reporter and the appearance of phenotypic defects overlapping those observed upon examination of worms with lesions in Dicer (dcr 1) or Drosha (drsh-1). Considered together, these results indicate a role for Pasha in miRNA maturation and miRNA-mediated gene regulation. PMID- 15531880 TI - Cytoplasmic ubiquitin ligase KPC regulates proteolysis of p27(Kip1) at G1 phase. AB - The cyclin-dependent kinase inhibitor p27(Kip1) is degraded at the G0-G1 transition of the cell cycle by the ubiquitin-proteasome pathway. Although the nuclear ubiquitin ligase (E3) SCF(Skp2) is implicated in p27(Kip1) degradation, proteolysis of p27(Kip1) at the G0-G1 transition proceeds normally in Skp2(-/-) cells. Moreover, p27(Kip1) is exported from the nucleus to the cytoplasm at G0-G1 (refs 9-11). These data suggest the existence of a Skp2-independent pathway for the degradation of p27(Kip1) at G1 phase. We now describe a previously unidentified E3 complex: KPC (Kip1 ubiquitination-promoting complex), consisting of KPC1 and KPC2. KPC1 contains a RING-finger domain, and KPC2 contains a ubiquitin-like domain and two ubiquitin-associated domains. KPC interacts with and ubiquitinates p27(Kip1) and is localized to the cytoplasm. Overexpression of KPC promoted the degradation of p27(Kip1), whereas a dominant-negative mutant of KPC1 delayed p27(Kip1) degradation. The nuclear export of p27(Kip1) by CRM1 seems to be necessary for KPC-mediated proteolysis. Depletion of KPC1 by RNA interference also inhibited p27(Kip1) degradation. KPC thus probably controls degradation of p27(Kip1) in G1 phase after export of the latter from the nucleus. PMID- 15531881 TI - Rate of molecular evolution of the seminal protein gene SEMG2 correlates with levels of female promiscuity. AB - Postcopulatory sperm competition is a key aspect of sexual selection and is believed to drive the rapid evolution of both reproductive physiology and reproduction-related genes. It is well-established that mating behavior determines the intensity of sperm competition, with polyandry (i.e., female promiscuity) leading to fiercer sperm competition than monandry. Studies in mammals, particularly primates, showed that, owing to greater sperm competition, polyandrous taxa generally have physiological traits that make them better adapted for fertilization than monandrous species, including bigger testes, larger seminal vesicles, higher sperm counts, richer mitochondrial loading in sperm and more prominent semen coagulation. Here, we show that the degree of polyandry can also impact the dynamics of molecular evolution. Specifically, we show that the evolution of SEMG2, the gene encoding semenogelin II, a main structural component of semen coagulum, is accelerated in polyandrous primates relative to monandrous primates. Our study showcases the intimate relationship between sexual selection and the molecular evolution of reproductive genes. PMID- 15531882 TI - Comparison of genome degradation in Paratyphi A and Typhi, human-restricted serovars of Salmonella enterica that cause typhoid. AB - Salmonella enterica serovars often have a broad host range, and some cause both gastrointestinal and systemic disease. But the serovars Paratyphi A and Typhi are restricted to humans and cause only systemic disease. It has been estimated that Typhi arose in the last few thousand years. The sequence and microarray analysis of the Paratyphi A genome indicates that it is similar to the Typhi genome but suggests that it has a more recent evolutionary origin. Both genomes have independently accumulated many pseudogenes among their approximately 4,400 protein coding sequences: 173 in Paratyphi A and approximately 210 in Typhi. The recent convergence of these two similar genomes on a similar phenotype is subtly reflected in their genotypes: only 30 genes are degraded in both serovars. Nevertheless, these 30 genes include three known to be important in gastroenteritis, which does not occur in these serovars, and four for Salmonella translocated effectors, which are normally secreted into host cells to subvert host functions. Loss of function also occurs by mutation in different genes in the same pathway (e.g., in chemotaxis and in the production of fimbriae). PMID- 15531883 TI - Induced recruitment of NK cells to lymph nodes provides IFN-gamma for T(H)1 priming. AB - Naive T cells are stimulated by antigen-presenting dendritic cells (DCs) in secondary lymphoid organs, but whether other types of cell participate in T cell priming is unclear. Here we show in mice that natural killer (NK) cells, which are normally excluded from lymph nodes, are rapidly recruited in a CCR7 independent, CXCR3-dependent manner to lymph nodes on stimulation by the injection of mature DCs. Recruitment of NK cells is also induced by some, but not all, adjuvants and correlates with the induction of T helper cell type 1 (T(H)1) responses. NK cell depletion and reconstitution experiments show that NK cells provide an early source of interferon-gamma (IFN-gamma) that is necessary for T(H)1 polarization. Taken together, our results identify an induced pathway of NK cell migration in antigen-stimulated lymph nodes and a mechanism by which some adjuvants may facilitate T(H)1 responses. PMID- 15531884 TI - An evolutionarily conserved target motif for immunoglobulin class-switch recombination. AB - Immunoglobulin H class-switch recombination (CSR) occurs between switch regions and requires transcription and activation-induced cytidine deaminase (AID). Transcription through mammalian switch regions, because of their GC-rich composition, generates stable R-loops, which provide single-stranded DNA substrates for AID. However, we show here that the Xenopus laevis switch region S(mu), which is rich in AT and not prone to form R-loops, can functionally replace a mouse switch region to mediate CSR in vivo. X. laevis S(mu)-mediated CSR occurred mostly in a region of AGCT repeats targeted by the AID-replication protein A complex when transcribed in vitro. We propose that AGCT is a primordial CSR motif that targets AID through a non-R-loop mechanism involving an AID replication protein A complex. PMID- 15531885 TI - Bone marrow plasmacytoid dendritic cells can differentiate into myeloid dendritic cells upon virus infection. AB - Two subsets of dendritic cell (DCs), plasmacytoid (p) and myeloid (m) DCs, have been described in humans and mice. These subsets are known to have divergent roles during an immune response, but their developmental course is unclear. Here we report that virus infection induces bone marrow pDCs to differentiate into mDCs, thereby undergoing profound phenotypic and functional changes including the acquisition of enhanced antigen-presenting capacity and the ability to recognize different microbial structures through Toll-like receptor 4. The conversion of pDCs into mDCs is also induced by the injection of double-stranded RNA and requires type I interferons. Our results establish a precursor-product developmental relationship between these two DC subsets and highlight unexpected plasticity of bone marrow pDCs. PMID- 15531886 TI - Sealing is at the origin of rubber slipping on wet roads. AB - Loss of braking power and rubber skidding on a wet road is still an open physics problem, as neither the hydrodynamic effects nor the loss of surface adhesion that are sometimes blamed really manage to explain the 20-30% observed loss of low-speed tyre-road friction. Here we report a novel mechanism based on sealing of water-filled substrate pools by the rubber. The sealed-in water effectively smoothens the substrate, thus reducing the viscoelastic dissipation in bulk rubber induced by surface asperities-well established as a major friction contribution. Starting with the measured spectrum of asperities one can calculate the water-smoothened spectrum and from that the predicted friction reduction, which is of the correct magnitude. The theory is directly supported by fresh tyre asphalt friction data. PMID- 15531887 TI - Manipulation of liquid droplets using amphiphilic, magnetic one-dimensional photonic crystal chaperones. AB - The controlled manipulation of small volumes of liquids is a challenging problem in microfluidics, and it is a key requirement for many high-throughput analyses and microassays. One-dimensional photonic crystals made from porous silicon have been constructed with amphiphilic properties. When prepared in the form of micrometre-sized particles and placed in a two-phase liquid such as dichloromethane/water, these materials will accumulate and spontaneously align at the interface. Here we show that superparamagnetic nanoparticles of Fe(3)O(4) can be incorporated into the porous nanostructure, allowing the materials to chaperone microlitre-scale liquid droplets when an external magnetic field is applied. The optical reflectivity spectrum of the photonic crystal displays a peak that serves to identify the droplet. Two simple microfluidics applications are demonstrated: filling and draining a chaperoned droplet, and combining two different droplets to perform a chemical reaction. The method provides a general means for manipulating and monitoring small volumes of liquids without the use of pumps, valves or a microfluidic container. PMID- 15531888 TI - Excitations of incoherent spin-waves due to spin-transfer torque. AB - The possibility of exciting microwave oscillations in a nanomagnet by a spin polarized current, as predicted by Slonczewski and Berger, has recently been demonstrated. This observation opens important prospects of applications in radiofrequency components. However, some unresolved inconsistencies are found when interpreting the magnetization dynamics within the coherent spin-torque model. In some cases, the telegraph noise caused by spin-currents could not be quantitatively described by that model. This has led to controversy about the need for an effective magnetic temperature model. Here we interpret the experimental results of Kiselev et al. using micromagnetic simulations. We point out the key role played by incoherent spin-wave excitation due to spin-transfer torque. The incoherence is caused by spatial inhomogeneities in local fields generating distributions of local precession frequencies. We observe telegraph noise with gigahertz frequencies at zero temperature. This is a consequence of the chaotic dynamics and is associated with transitions between attraction wells in phase space. PMID- 15531889 TI - Berberine is a novel cholesterol-lowering drug working through a unique mechanism distinct from statins. AB - We identify berberine (BBR), a compound isolated from a Chinese herb, as a new cholesterol-lowering drug. Oral administration of BBR in 32 hypercholesterolemic patients for 3 months reduced serum cholesterol by 29%, triglycerides by 35% and LDL-cholesterol by 25%. Treatment of hyperlipidemic hamsters with BBR reduced serum cholesterol by 40% and LDL-cholesterol by 42%, with a 3.5-fold increase in hepatic LDLR mRNA and a 2.6-fold increase in hepatic LDLR protein. Using human hepatoma cells, we show that BBR upregulates LDLR expression independent of sterol regulatory element binding proteins, but dependent on ERK activation. BBR elevates LDLR expression through a post-transcriptional mechanism that stabilizes the mRNA. Using a heterologous system with luciferase as a reporter, we further identify the 5' proximal section of the LDLR mRNA 3' untranslated region responsible for the regulatory effect of BBR. These findings show BBR as a new hypolipidemic drug with a mechanism of action different from that of statin drugs. PMID- 15531890 TI - Specific peptide interference reveals BCL6 transcriptional and oncogenic mechanisms in B-cell lymphoma cells. AB - The BTB/POZ transcriptional repressor and candidate oncogene BCL6 is frequently misregulated in B-cell lymphomas. The interface through which the BCL6 BTB domain mediates recruitment of the SMRT, NCoR and BCoR corepressors was recently identified. To determine the contribution of this interface to BCL6 transcriptional and biological properties, we generated a peptide that specifically binds BCL6 and blocks corepressor recruitment in vivo. This inhibitor disrupts BCL6-mediated repression and establishment of silenced chromatin, reactivates natural BCL6 target genes, and abrogates BCL6 biological function in B cells. In BCL6-positive lymphoma cells, peptide blockade caused apoptosis and cell cycle arrest. BTB domain peptide inhibitors may constitute a novel therapeutic agent for B-cell lymphomas. PMID- 15531891 TI - Role of ISG15 protease UBP43 (USP18) in innate immunity to viral infection. AB - Innate immune responses provide the host with an early protection barrier against infectious agents, including viruses, and help shape the nature and quality of the subsequent adaptive immune responses of the host. Expression of ISG15 (UCRP), a ubiquitin-like protein, and protein ISGylation are highly increased upon viral infection. We have identified UBP43 (USP18) as an ISG15 deconjugating protease. Protein ISGylation is enhanced in cells deficient in UBP43 (ref. 6). Here we have examined the role of UBP43, encoded by the gene Usp18, in innate immunity to virus infection. Usp18(-/-) mice were resistant to the fatal lymphocytic choriomeningitis and myeloencephalitis that developed in wild-type mice after intracerebral inoculation with lymphocytic choriomeningitis virus (LCMV) or vesicular stomatitis virus (VSV), respectively. Survival of Usp18(-/-) mice after intracerebral LCMV infection correlated with a severe inhibition of LCMV RNA replication and antigen expression in the brain and increased levels of protein ISGylation. Consistent with these findings, mouse embryonic fibroblasts (MEF) and bone marrow-derived macrophages from Usp18(-/-) mice showed restricted LCMV replication. Moreover, MEF from Usp18(-/-) mice showed enhanced interferon mediated resistance to the cytopathic effect caused by VSV and Sindbis virus (SNV). This report provides the first direct evidence that the ISG15 protease UBP43 and possibly protein ISGylation have a role in innate immunity against viral infection. PMID- 15531893 TI - Cell cycle-dependent regulation of yeast telomerase by Ku. AB - The heterodimeric Ku complex affects telomere structure in diverse organisms. We report here that in the absence of Ku, the catalytic subunit of telomerase, Est2p, was not telomere-associated in G1 phase, and its association in late S phase was decreased. The telomere association of Est1p, a telomerase component that binds telomeres only in late S phase, was also reduced in the absence of Ku. The effects of Ku on telomerase binding require a 48-nucleotide (nt) stem-loop region of TLC1 telomerase RNA. Ku interacts with TLC1 RNA via this 48-nt region throughout the cell cycle, but this interaction was reduced after telomere replication. These data support a model in which Ku recruits telomerase to telomeres in G1 phase when telomerase is inactive and promotes telomerase mediated telomere lengthening in late S phase. PMID- 15531892 TI - Escherichia coli Hfq has distinct interaction surfaces for DsrA, rpoS and poly(A) RNAs. AB - The bacterial Sm-like protein Hfq facilitates RNA-RNA interactions involved in post-transcriptional regulation of the stress response. Specifically, Hfq helps pair noncoding RNAs (ncRNAs) with complementary regions of target mRNAs. To probe the mechanism of this pairing, we generated a series of Hfq mutants and measured their affinity for RNAs like those with which Hfq must associate in vivo. We tested the mutants' DsrA-dependent activation of rpoS, and their ability to stabilize DsrA ncRNA against degradation in vivo. Our results suggest that Hfq has two independent RNA-binding surfaces. In addition to a well-known site around the core of the Hfq hexamer, we observe interactions with the distal face of Hfq, a new locus with which mRNAs and poly(A) sequences associate. Our model explains how Hfq can simultaneously bind a ncRNA and its mRNA target to facilitate the strand displacement reaction required for Hfq-dependent translational regulation. PMID- 15531894 TI - Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction. AB - Allogeneic bone marrow transplantation (BMT) with marrow ablative conditioning is the treatment of choice for haematopoietic malignancies. The use of nonmyeloablative stem cell transplants has allowed the treatment of patients previously ineligible for BMT because of age or other disease. These reduced conditioning regimes allow the persistence initially of some recipient cells in the blood and bone marrow (haematopoietic chimaerism). Monitoring of the relative proportion of donor and recipient cells is required to assess the success of the procedure, to predict subsequent rejection or impending relapse and to guide the use of donor lymphocyte infusions. We present a quantitative real-time PCR approach for the measurement of haematopoietic chimaerism using the TaqMan. This approach exploits the presence of single-nucleotide polymorphisms (SNPs) to distinguish cells of patient or donor origin. We have designed and validated a panel of seven allele-specific probes to quantify the contribution of patient and donor cells in the haematopoietic population from 12 patient and donor pairs. We have compared the performance of this approach with an existing method and proved it to be superior in both accuracy and sensitivity. The use of more sensitive and accurate techniques permits earlier intervention for improved clinical outcome. PMID- 15531895 TI - Thalidomide salvage therapy following allogeneic stem cell transplantation for multiple myeloma: a retrospective study from the Intergroupe Francophone du Myelome (IFM) and the Societe Francaise de Greffe de Moelle et Therapie Cellulaire (SFGM-TC). AB - Thalidomide is effective in multiple myeloma (MM), even in patients who have relapsed after high-dose therapy. A potent graft-versus-myeloma (GVM) effect can be induced against MM after allogeneic stem cell transplantation (allo-SCT). In all, 31 MM patients received thalidomide as a salvage therapy after progression following allo-SCT. The median maximum daily dose of thalidomide was 200 mg (range, 50-600). Thalidomide had to be discontinued in six patients (19%) because of toxicity. In all, nine patients (29%; 95% CI, 13-45) achieved an objective response with thalidomide therapy (six partial and three very good partial responses, VGPR). Five patients developed graft-versus-host disease (GVHD) after thalidomide therapy, including the three patients achieving a VGPR. These data demonstrate that thalidomide is potentially effective in MM patients failing allo SCT. PMID- 15531896 TI - Outcomes of transplantation with partial T-cell depletion of matched or mismatched unrelated or partially matched related donor bone marrow in children and adolescents with leukemias. AB - Graft-versus-host disease (GVHD) remains a major barrier to successful hematopoietic stem cell transplant for patients who lack a matched related donor. Partial T-cell depletion (TCD) of the graft may decrease the risk of severe GVHD with unrelated donors (URD) and partially matched related donors (PMRD) while retaining an antileukemic effect. We analyzed our experience using URD and PMRD for pediatric patients with leukemias from 1990 to 2001. A subgroup of 'matched' URD donor pairs was retrospectively analyzed for high-resolution class I. Partial TCD was accomplished with monoclonal antibody T10B9 or OKT3 and complement. There were 76 URD (45% matched) and 28 PMRD recipients. Event-free survival (EFS) was 38.3%, and overall survival (OS) 45.1% at 3 years. On multivariate analysis, there was no difference in survival based upon marrow source, but nonrelapse mortality was higher with the use of PMRD. Relapse occurred in 6% of ALL patients, and 22.8% of AML/MDS patients. Grades III-IV GVHD was observed in only 6.7% of patients. Partial TCD allows use of matched or mismatched URD, or PMRD with little mortality from GVHD, durable engraftment, and no increase in relapse risk. PMID- 15531897 TI - Impact of early NK cell recovery on development of GvHD and CMV reactivation in dose-reduced regimen prior to allogeneic PBSCT. AB - Dose-reduced allogeneic peripheral blood stem cell transplantation (PBSCT) is a therapeutic approach for patients with haematological malignancies who are not eligible for conventional allogeneic PBSCT. We analysed early development of lymphocyte subpopulations and the occurrence of cytomegalovirus (CMV) reactivation and acute graft-versus-host reaction (GvHD) in patients undergoing the protocol according to Slavin vs conventionally treated patients. Lymphocyte status prior to conditioning and at day +30 after allogeneic PBSCT was determined in 24 out of 51 patients who received conventional allogeneic PBSCT (eg cyclophosphamide plus total body irradiation) and compared with 27 patients being treated according to the Slavin protocol (fludarabine, busulphan and ATG). There is a significant delay in CD4 (T helper) cell development and consecutive lower CD4/CD8 ratios and a better reconstitution of CD8 (T cytotoxic) and NK (natural killer) cells after the Slavin protocol. Patients undergoing this protocol and no, or only grade I, acute GvHD show an even better NK cell reconstitution compared to patients with grade II-IV GvHD. A low CD4/CD8 ratio represents a CMV risk factor only in conventionally treated patients with grade 0-I GvHD, while after preparative regimen according to the Slavin protocol, the NK/CD8 ratio might be a marker for the prediction of CMV reactivation in addition to CMV risk status. PMID- 15531898 TI - Life-threatening neurological complications after bone marrow transplantation in children. AB - Neurological complications may occur in BMT recipients (11-59%), frequently contributing to morbidity or mortality. They are the main causes of death in 10 15%. Life-threatening neurological complications were seen in 11 out of 113 (9.7%) children who underwent BMT from HLA-matched family (n=7) or mismatched donors (n=4) at our institution. Diagnoses of patients with neurological complications were acute myeloblastic leukemia (AML) (five), thalassemia major (two), Fanconi anemia (two), Omenn syndrome (one) and leukodystrophy (one), and the neurological events were seen between days +13 and +85 after transplantation. Minor symptoms including reversible, nonrepetitive seizures were excluded. Cyclosporine A toxicity was diagnosed in six children. The rest of the complications were brain abscess/meningoencephalitis (two), severe hypomagnesemia (one), busulfan toxicity (one), sustained hypertension (three), and intracranial hemorrhage (three). Six patients with neurological complications suffered from >grade II graft-versus-host disease (GvHD), and all were high risk for transplant related complications. In this study, risk status of the underlying disease, mismatched transplantation, a diagnosis of AML (advanced stage), older age and >grade II GvHD were important adverse factors for the development of severe life threatening neurological complications. PMID- 15531899 TI - Correction of recurrent angioedema related to C1 esterase inhibitor deficiency as a secondary event following nonmyeloablative allogeneic stem cell transplantation in a patient with myelofibrosis. PMID- 15531900 TI - Sinus bradycardia associated with cyclosporine following allogeneic hematopoietic stem cell transplantation. PMID- 15531901 TI - Hematopoietic stem cell transplantation for multiply transfused patients with sickle cell disease and thalassemia after low-dose total body irradiation, fludarabine, and rabbit anti-thymocyte globulin. AB - Patients with sickle cell disease (N = 3) and thalassemia (N = 1) with high-risk features received hematopoietic stem cell transplantations (HCT) to induce stable (full or partial) donor engraftment. Patients were 9-30 years of age. Fludarabine, rabbit anti-thymocyte globulin (ATG), and 200 cGy total body irradiation were administered pre-transplant. Patients received bone marrow (N = 3) or peripheral blood stem cells (N = 1) from HLA-identical siblings, followed by mycophenolate mofetil and cyclosporine for post-grafting immunosuppression. Significant lymphopenia, but only moderate neutropenia and thrombocytopenia developed post transplant. No grade IV nonhematological toxicities or acute graft versus-host disease (GVHD) were observed. At 3 months after transplantation, three of four patients had evidence of donor myeloid chimerism (range, 15-100%). However, after post transplant immunosuppression was discontinued, graft rejection occurred in all but one patient. This patient is now doing well 27 months post transplant with full donor engraftment. One patient died after a second transplant, and another patient experienced a stroke as her graft was being rejected. These results suggest that stable donor engraftment after nonmyeloablative HCT is difficult to achieve among immunocompetent patients with hemoglobinopathies and that new approaches will need to be developed before wider application of this transplantation method for hemoglobinopathies. PMID- 15531902 TI - Maintenance rituximab after autologous stem cell transplant for high-risk B-cell lymphoma induces prolonged and severe hypogammaglobulinemia. PMID- 15531903 TI - Impact of HLA class I high-resolution mismatches on chronic graft-versus-host disease and survival of patients given hematopoietic stem cell grafts from unrelated donors. AB - There is consensus that matching of unrelated donors (URD) and patients for HLA class II alleles improves the outcome of hematopoietic stem cell transplantation (HSCT). However, the significance of HLA class I allelic mismatches for transplant outcome is under discussion and reports on long-term effects like chronic graft-versus-host disease (GVHD) are rare. Thus, we investigated the association of human leukocyte antigen (HLA) class I allele mismatches and outcome in 144 patients given HSCT from URD who were matched for HLA-DRB1, DRB3/4/5, and DQB1 alleles. The risk of chronic GVHD was significantly increased in patients with class I mismatched donors, the mismatch either detected by low- or high-resolution typing. A single HLA class I allele mismatch significantly increased the risk of chronic GVHD in multivariate analysis. Overall survival was significantly reduced in patient/donor pairs with more than one-allele class I mismatch. Thus, selection of unrelated donors for transplantation should be based on high-resolution HLA class I typing. PMID- 15531904 TI - Severe hypercholesterolemia mediated by lipoprotein X in patients with chronic graft-versus-host disease of the liver. AB - We describe a series of cases of extreme hypercholesterolemia mediated by lipoprotein X in patients with chronic graft-versus-host disease of the liver after an allogeneic bone marrow transplant. All of the patients presented with a total cholesterol in excess of 1000 mg/dl (25.9 mmol/l). At the time they were also noted to have pseudohyponatremia. Cholesterol appeared to be predominantly carried by lipoprotein X. Intrahepatic cholestasis leading to reflux of bile lipoproteins into the bloodstream and subsequent formation of protein X appears to be the mechanism underlying this phenomenon. Complications, including retinal cholesterol thromboembolism and cholesteroloma of the lung have been seen in the patient with the highest cholesterol levels. Severe hypercholesterolemia is an important, and likely more common than previously reported, long-term complication of allogeneic hematopoietic stem cell transplantation. It is important for clinicians to familiarize themselves with the diagnostic and therapeutic challenges this condition presents. PMID- 15531905 TI - Venous thromboembolism after high-dose chemotherapy in a patient with Hodgkin's lymphoma receiving the new oral contraceptive ethinylestradiol and drospirenone ("Yasmine"). PMID- 15531906 TI - Successful transplantation of peripheral blood stem cells mobilized by chemotherapy and a single dose of pegylated G-CSF in patients with multiple myeloma. AB - Following induction therapy and 4 g/m(2) cyclophosphamide, a single dose of 12 mg polyethyleneglycol-conjugated G-CSF (pegfilgrastim; n=12) or daily doses of unconjugated G-CSF (8.5 mug/kg/day) (n=12) were administered to myeloma patients. Pegfilgrastim was associated with an earlier leukocyte recovery (12 vs 14 days) and peripheral blood CD34+ cell peak (12 vs 15 days). The peripheral blood CD34+ cell peak was lower in the pegfilgrastim group (78 vs 111/mul). Following high dose melphalan (200 mg/m(2)) and autografting, leukocyte and platelet reconstitution was similar in both groups and stable blood counts were observed 100 days post transplant. In summary, a single dose of pegfilgrastim after chemotherapy is capable of mobilizing a sufficient number of CD34+ cells for successful autografting with early engraftment and sustained hematological reconstitution in patients with myeloma. These data provide the basis for randomized studies evaluating the optimal dose and time of pegfilgrastim as well as long-term outcome in larger cohorts of patients. PMID- 15531907 TI - Stem cell transplantation from identical twins in patients with myelodysplastic syndromes. AB - In a multicentre retrospective EBMT database study, we analysed factors influencing outcome in 38 patients with MDS/sAML who were transplanted with stem cells from their syngeneic twin and compared those to 1444 patients who were transplanted from an HLA-identical sibling. The median time to leukocyte and platelet engraftment was faster in the twin group: 14 vs 17 (P=0.02) and 16 vs 26 days (P=0.09), respectively. The 5 years cumulative incidence of treatment related mortality (TRM) was higher in the sibling than in the twin group (38 vs 27%; P=0.05). The 5 year cumulative incidence of relapse was 32% (95% CI: 29-35%) for the siblings and 39% (95% CI: 26-60%; P=0.6) for the twins. A trend for better 5-years disease-free and overall survival was observed in the twin group: 34% (95% CI: 14-54%) vs 28% (95% CI: 25-31%; P=0.2) and 36% (95% CI: 15-57%) vs 32% (95% CI: 29-35%; P=0.09), respectively. In a multivariate analysis, stem cell transplantation from identical twins had a lower TRM: HR: 0.4 (95% CI: 0.2-0.9; P=0.03). The relapse rate was similar for both groups with a HR of 1.2 (95% CI: 0.07-2.1; P=0.5), with a better survival for the twins: HR 0.6 (95% CI: 0.4-1.0; P=0.07). We conclude that twin transplantation in MDS/sAML is associated with a similar relapse risk, a lower TRM and a trend for better overall survival in comparison to transplantation from HLA-identical siblings. PMID- 15531908 TI - Camostat mesilate attenuates pancreatic fibrosis via inhibition of monocytes and pancreatic stellate cells activity. AB - Camostat mesilate (CM), an oral protease inhibitor, has been used clinically for the treatment of chronic pancreatitis in Japan. However, the mechanism by which it operates has not been fully understood. Our aim was to evaluate the therapeutic efficacy of CM in the experimental pancreatic fibrosis model induced by dibutyltin dichloride (DBTC), and we also determined the effect of CM on isolated monocytes and panceatic stellate cells (PSCs). In vivo, chronic pancreatitis was induced in male Lewis rats by single administration of 7 mg/kg DBTC and a special diet containing 1 mg/g CM was fed to the DBTC+CM-treated group from day 7, while the DBTC-treated group rats were fed a standard diet. At days 0, 7, 14 and 28, the severity of pancreatitis and fibrosis was examined histologically and enzymologically in both groups. In vitro, monocytes were isolated from the spleen of a Lewis rat, and activated with lipopolysaccharide stimulation. Thereafter, the effect of CM on monocyte chemoattractant protein-1 (MCP-1) and tumor necrosis factor-alpha (TNF-alpha) production from monocytes was examined. Subsequently, cultured rat PSCs were exposed to CM and tested to see whether their proliferation, MCP-1 production and procollagen alpha1 messenger RNA expression was influenced by CM. In vivo, the oral administration of CM inhibited inflammation, cytokines expression and fibrosis in the pancreas. The in vitro study revealed that CM inhibited both MCP-1 and TNF-alpha production from monocytes, and proliferation and MCP-1 production from PSCs. However, procollagen alpha1 expression in PSCs was not influenced by CM. These results suggest that CM attenuated DBTC-induced rat pancreatic fibrosis via inhibition of monocytes and PSCs activity. PMID- 15531909 TI - Microsomal prostaglandin E synthase (mPGES)-1, mPGES-2 and cytosolic PGES expression in human gastritis and gastric ulcer tissue. AB - Recently, three different prostaglandin E2 synthases have been identified: microsomal prostaglandin E synthase (mPGES)-1, cytosolic PGES (cPGES), and mPGES 2; however, their role and connection to cyclooxygenase (COX)-2 in the gastric ulcer repair process remain unknown. Therefore, we examined mPGES-1, cPGES, and mPGES-2 expression and localization in the stomach in vitro and in vivo. Tissues were obtained from Helicobacter pylori (H. pylori)-infected patients and consisted of surgical resections of gastric ulcers, or biopsies of gastric ulcers or gastritis. mPGES-1 mRNA and protein expression levels were examined by real time polymerase chain reaction (PCR) and Western blot analysis, respectively. mPGES-1, cPGES, and mPGES-2 localization were analyzed immunohistochemically. Induction of PGES expression in response to interleukin (IL)-1beta was examined in vitro in the cultured human gastric fibroblast line Hs262.St. Real-time PCR analysis of mPGES-1 mRNA expression in biopsy samples showed significantly higher expression levels in open than in closed gastric ulcer tissue. Western blot analysis showed mPGES-1 protein expression limited to open ulcer tissue, while mPGES-2 and cPGES immunoreactivities were seen in both open and closed ulcer tissue. Immunohistochemical analysis showed strong mPGES-1 expression in fibroblasts and macrophages of the ulcer bed, paralleling COX-2 expression. cPGES and mPGES-2 expression levels were seen in both fibroblasts of the ulcer bed and in epithelial cells. Furthermore, stronger cPGES and mPGES-2 immunoreactivities were seen in scattered mast cell-like cells and neuroendocrine-like cells, respectively. Induction of mPGES-1 expression in response to IL-1beta was seen in cultured gastric fibroblasts in vitro, and double immunostaining showed mPGES-1 coexpression with COX-2 in fibroblasts of the ulcer bed in vivo. In conclusion, mPGES-1, cPGES, and mPGES-2 are all expressed in gastric ulcer tissue, but only mPGES-1 parallels COX-2 expression in mesenchymal and inflammatory cells of the ulcer bed, suggesting a key role for this enzyme in the ulcer repair process. PMID- 15531910 TI - Ral-GTPases: good chances for a long-lasting fame. PMID- 15531911 TI - Conditional E2F1 activation in transgenic mice causes testicular atrophy and dysplasia mimicking human CIS. AB - E2F1 is a crucial downstream effector of the retinoblastoma protein (pRB) pathway. To address the consequences of short-term increase in E2F1 activity in adult tissues, we generated transgenic mice expressing the human E2F1 protein fused to the oestrogen receptor (ER) ligand-binding domain. The expression of the ER-E2F1 fusion protein, which is inactive in the absence of 4-hydroxy tamoxifen (OHT), was targeted to the testes. We show that short-term activation of E2F1 results in activation of E2F target genes and apoptosis of germ cells. Consistent with our previously published results, the apoptotic response was independent of p53. Persistent E2F1 activation for 3 weeks led to massive apoptosis and severe testicular atrophy with seminiferous tubules containing only Sertoli cells and clusters of undifferentiated spermatogonia. The latter showed high expression of ER-E2F1 and excessive mitotic activity, including atypical mitoses. In addition, gonocyte-like dysplastic germ cells, resembling carcinoma in situ (CIS) cells in humans, appeared. Our results show that a relatively short period of deregulated E2F1 activity in testicles can induce premalignant changes. Moreover, we demonstrate the feasibility of tissue-specific expression of conditional ER-E2F1 in transgenic mice. PMID- 15531912 TI - Caspase-8 gene is frequently inactivated by the frameshift somatic mutation 1225_1226delTG in hepatocellular carcinomas. AB - Evidence exists that alterations of the genes encoding apoptosis-related proteins contribute to either development or progression of human cancers. Caspase-8 plays a crucial role in the initiation phase of apoptosis. To explore the possibility that the genetic alteration of caspase-8 gene is involved in the development of hepatocellular carcinomas (HCCs), we have analysed the entire coding region of human caspase-8 gene for the detection of somatic mutations by polymerase chain reaction-single-strand conformation polymorphism in 69 HCCs with low-grade dysplastic nodule (LGDN, n=2) or high-grade dysplastic nodule (HGDN, n=2) or without any dysplastic nodules (n=65). Overall, we detected a total of nine somatic mutations in 69 HCCs (13.0%). Interestingly, all of the nine mutations were an identical frameshift mutation with two base-pair deletion (1225_1226delTG), which would result in a premature termination of amino-acid synthesis in the p10 protease subunit. In a patient sample, we detected the 1225_1226delTG mutation both in HCC and LDGN lesions, suggesting that caspase-8 mutation could be involved in the early stage of HCC carcinogenesis. We expressed the tumor-derived caspase-8 mutant in the cells and found that the mutant abolished cell death activity of caspase-8. Our data indicate that caspase-8 gene is frequently mutated in HCC and the majority of the mutations may be the frameshift mutation 1225_1226delTG. Also, the data suggest that caspase-8 gene mutation might lead to the loss of its cell death function and contribute to the pathogenesis of HCC. PMID- 15531913 TI - Rottlerin sensitizes glioma cells to TRAIL-induced apoptosis by inhibition of Cdc2 and the subsequent downregulation of survivin and XIAP. AB - In the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-resistant glioma cells, treatment with TRAIL in combination with subtoxic doses of rottlerin induced rapid apoptosis. While the proteolytic processing of procaspase 3 by TRAIL was partially blocked in these cells, treatment with rottlerin efficiently recovered TRAIL-induced activation of caspases. Treatment with rottlerin significantly decreased Cdc2 activity through the downregulation of cyclin A, cyclin B, and Cdc2 proteins, whereas the sensitizing effect of rottlerin on TRAIL-induced apoptosis was independent of PKCdelta activity. Furthermore, treatment with rottlerin downregulated the protein levels of survivin and X-chromosome-linked IAP (XIAP), two major caspase inhibitors. Forced expression of Cdc2 together with cyclin B attenuated rottlerin-potentiated TRAIL induced apoptosis by over-riding the rottlerin-mediated downregulation of survivin and XIAP protein levels. Taken together, inhibition of Cdc2 activity and the subsequent downregulation of survivin and XIAP by subtoxic doses of rottlerin contribute to amplification of caspase cascades, thereby overcoming resistance of glioma cells to TRAIL-mediated apoptosis. Since rottlerin can sensitize Bcl-2- or Bcl-xL-overexpressing glioma cells but not human astrocytes to TRAIL-induced apoptosis, this combined treatment may offer an attractive strategy for safely treating resistant gliomas. PMID- 15531914 TI - Smad4 deficiency in cervical carcinoma cells. AB - Squamous cell carcinoma of the uterine cervix is one of the most frequent cancers affecting women worldwide. Carcinomas arise from cervical intraepithelial lesions, in which infection with high-risk human papillomavirus types has led to deregulated growth control through the actions of the viral E6 and E7 oncoproteins. The molecular mechanisms underlying progression to invasive tumor growth are poorly understood. One important feature, however, is the escape from growth inhibition by transforming growth factor beta (TGF-beta). Loss of chromosomal arm 18q is among the most frequent cytogenetic alterations in cervical cancers and has been associated with poor prognosis. Since the TGF-beta response is mediated by Smad proteins and the tumor suppressor gene Smad4 resides at 18q21, we have analysed the Smad4 gene for cervical cancer-associated alterations in cell lines and primary carcinomas. Here, we report Smad4 deficiency in four out of 13 cervical cancer cell lines which is due to an intronic rearrangement or deletions of 3' exons. All cell lines, however, showed either absent or moderate responsiveness to TGF-beta irrespective of their Smad4 status. In 41 primary squamous cervical carcinomas analysed, 10 samples showed loss of Smad4 protein expression and 26 samples a reduced expression. Altogether, our results strongly suggest that Smad4 gene alterations are involved in cervical carcinogenesis. PMID- 15531915 TI - Ets1 is an effector of protein kinase Calpha in cancer cells. AB - PKCalpha and Ets1 are both associated with breast cancer progression. Our previous studies suggested that these proteins are likely to functionally interact with one another. Here, we show that attenuation of endogenous PKCalpha expression (siPalpha) by RNA interference leads to reduced Ets1 protein expression in a variety of cancer cells. Pulse-chase experiments and treatment with proteasome inhibitor MG-132 revealed that siPalpha interferes with both Ets1 protein synthesis and stability. The effect of siPalpha on Ets1 expression could be partially prevented by KN-93, suggesting that calcium/calmodulin-dependent kinase II (CaMKII), a modulator of Ets1 activity, may play a role in PKCalpha dependent Ets1 regulation. In contrast, Ets1-regulating kinases ERK1/2 were not found to be involved in this process. To assess the importance of the PKCalpha/Ets1 interaction, we compared the biological responses of MDA-MB-231 cells to PKCalpha- and Ets1-specific siRNAs (siE1). While only siPalpha induced changes in cellular morphology and anchorage-independent growth, both siRNAs similarly affected cellular responses to the antitumor drug mithramycin A and to UV light. Microarray analyses further showed that the expression of a certain set of genes was equally affected by siPalpha and siE1. The data suggest that Ets1 serves as an effector for PKCalpha to fulfil certain functions in cancer cells. PMID- 15531916 TI - Ron tyrosine kinase receptor regulates papilloma growth and malignant conversion in a murine model of skin carcinogenesis. AB - Recent studies demonstrate that the receptor tyrosine kinase (TK) Ron is tumorigenic when overexpressed and plays a role in regulating skin homeostasis. We hypothesized that Ron signaling promotes skin carcinogenesis. To test this hypothesis, mice deficient in the TK domain of Ron (TK(-/-) mice) were crossed with v-Ha-ras (Tg.AC) transgenic mice; the resulting TK(-/-) Tg.AC(+/-) mice, and their controls, were utilized in a model of chemically induced Ras-mediated skin carcinogenesis. The mice were treated with 2.5 microg of 12-O tetradecanoylphorbol-13-acetate applied weekly to the shaved back of 36 control (TK(+/+) Tg.AC(+/-)) and 35 experimental (TK(-/-) Tg.AC(+/-)) mice. In an analysis of the resulting papillomas, a reduction in cellular proliferation and papilloma volume was found in the TK(-/-) Tg.AC(+/-) mice compared to controls. Further, Ron protein expression was upregulated during papilloma formation. Ablation of Ron signaling resulted in partial defects in MAPK and Akt signaling that may account for the decreased papilloma growth in the TK(-/-) Tg.AC(+/-) mice. The papilloma-bearing mice were monitored for the occurrence of malignant skin tumors and other malignant tumor types for a period of 48 weeks. Loss of Ron receptor signaling significantly reduced the percent of papillomas that underwent malignant conversion as well as the number of mice developing other malignant tumor types. In conclusion, these studies demonstrate that Ron signaling augments papilloma growth and malignant conversion in vivo. PMID- 15531917 TI - Transcript profiling of Wilms tumors reveals connections to kidney morphogenesis and expression patterns associated with anaplasia. AB - Anaplasia (unfavorable histology) is associated with therapy resistance and poor prognosis of Wilms tumor, but the molecular basis for this phenotype is unclear. Here, we used a cDNA array with 9240 clones relevant to cancer biology and/or kidney development to examine the expression profiles of 54 Wilms tumors, five normal kidneys and fetal kidney. By linking genes differentially expressed between fetal kidney and Wilms tumors to kidney morphogenesis, we found that genes expressed at a higher level in Wilms tumors tend to be expressed more in uninduced metanephrogenic mesenchyme or blastema than in their differentiated structures. Conversely, genes expressed at a lower level in Wilms tumors tend to be expressed less in uninduced metanephrogenic mesenchyme or blastema. We also identified 97 clones representing 76 Unigenes or unclustered ESTs that clearly separate anaplastic Wilms tumors from tumors with favorable histology. Genes in this set provide insight into the nature of the abnormal nuclear morphology of anaplastic tumors and may facilitate identification of molecular targets to improve their responsiveness to treatment. PMID- 15531918 TI - Proteasome inhibitor PS-341 causes cell growth arrest and apoptosis in human glioblastoma multiforme (GBM). AB - The proteasome plays a pivotal role in controlling cell proliferation, apoptosis, and differentiation in a variety of normal and tumor cells. PS-341, a novel boronic acid dipeptide that inhibits 26S proteasome activity, has prominent effects in vitro and in vivo against several solid tumors. We examined its antiproliferation, proapoptotic effects using three human glioblastoma multiforme (GBM) cell lines and five primary GBM explants. PS-341 markedly inhibited proliferation of GBM cell lines and explants in liquid and soft agar culture. These cells developed a G2/M cell cycle arrest with a concomitant decreased percentage of cells in S phase ( approximately 2-fold), associated with an increased expression of p21(WAF1), p27(KIP1), as well as cyclin B1 and decreased levels of CDK2, CDK4, and E2F4. About 35-40% of the cells became apoptotic when exposed to PS-341 (10(-7) M, 24-48 h) as shown by Annexin V analysis; in concert with these findings, immunobloting showed a C-terminal 85 kDa apoptotic fragment of poly ADP-ribose polymerase (PARP), and a decreased level of Bcl2 and Bcl-xl. PS-341 downregulated the expression of Bcl-2 and Bcl-xl in protein levels at an early time of treatment. These changes occurred irrespective of the p53 mutational status of the cells. PS-341 activated JNK/c-Jun signaling in GBM cells, and the JNK inhibitor SP600125 blocked the JNK signaling to reverse partially the PS-341 growth inhibition. PS-341 (10(-7) M, 24 h) decreased nuclear NF-kappaB levels as shown by Western blot, and reduced transcriptional activity of NF-kappaB as measured by reporter assays in these transformed cells. Also, PS 341 enhanced TRAIL (TNF-related apoptosis-inducing ligand) and TNFalpha (tumor necrosis factor alpha) induced cell death and apoptosis (two- to five-fold) in GBM cells. In summary, PS-341 has profound effects on growth and apoptosis of GBM cells, suggesting that PS-341 may be an effective therapy for patients with gliomas. PMID- 15531919 TI - CuZnSOD deficiency leads to persistent and widespread oxidative damage and hepatocarcinogenesis later in life. AB - Mice deficient in CuZn superoxide dismutase (CuZnSOD) showed no overt abnormalities during development and early adulthood, but had a reduced lifespan and increased incidence of neoplastic changes in the liver. Greater than 70% of Sod1-/- mice developed liver nodules that were either nodular hyperplasia or hepatocellular carcinoma (HCC). Cross-sectional studies with livers collected from Sod1-/- and age-matched +/+ controls revealed extensive oxidative damage in the cytoplasm and, to a lesser extent, in the nucleus and mitochondria from as early as 3 months of age. A marked reduction in cytosolic aconitase, increased levels of 8-oxo dG and F2-isoprostanes, and a moderate reduction in glutathione peroxidase activities and porin levels were observed in all age groups of Sod1-/- mice examined. There were also age-related reductions in Mn superoxide dismutase activities and carbonic anhydrase III. Parallel to the biochemical changes, there were progressive increases in the DNA repair enzyme APEX1, the cell cycle control proteins cyclin D1 and D3, and the hepatocyte growth factor receptor Met. Increased cell proliferation in the presence of persistent oxidative damage to macromolecules likely contributes to hepatocarcinogenesis later in life. PMID- 15531920 TI - Array-CGH analysis of microsatellite-stable, near-diploid bowel cancers and comparison with other types of colorectal carcinoma. AB - Microsatellite-stable, near-diploid (MSI-CIN-) colorectal carcinomas have been reported, but it is not clear as to whether these tumours form a discrete group or represent one end of the distribution of MSI-CIN+ cancers. In order to address this question, we screened 23 MSI-CIN- colorectal cancers for gains and losses using array-based comparative genomic hybridization (aCGH) based on large-insert clones at about 1 Mb density. We compared our findings with those from a small set of MSI+CIN+ cancers, and with our reported data from MSI-CIN+ and MSI+CIN- cancers. We found no evidence of any form of genomic instability in MSI-CIN- cancers. At the level of the chromosome arm, the MSI-CIN- cancers had significantly fewer gains and losses than MSI-CIN+ tumours, but more than the MSI+CIN- and MSI+CIN+ lesions. The chromosomal-scale changes found in MSI-CIN- cancers generally involved the same sites as those in MSI-CIN+ tumours, and in both cancer groups, the best predictor of a specific change was the total number of such changes in that tumour. A few chromosomal-scale changes did, however, differ between the MSI-CIN- and MSI-CIN+ pathways. MSI-CIN- cancers showed: low frequencies of gain of 9p and 19p; infrequent loss of 5q and a high frequency of 20p gain. Overall, our data suggested that the MSI-CIN- group is heterogeneous, one type of MSI-CIN- cancer having few (< or =6) chromosomal-scale changes and the other with more (> or =10) changes resembling MSI-CIN+ cancers. At the level of individual clones, frequent and/or discrete gains or losses were generally located within regions of chromosomal-scale changes in both MSI-CIN- and MSI-CIN+ cancers, and fewer losses and gains were present in MSI-CIN- than MSI-CIN+ tumours. No changes by clone, which were specific to the MSI-CIN- cancers, were found. In addition to indicating differences among the cancer groups, our results also detected over 50 sites (amplifications, potential homozygous deletion and gains or losses which extended over only a few megabases) which might harbour uncharacterized oncogenes or tumour suppressor loci. In conclusion, our data support the suggestion that some MSI-CIN- carcinomas form a qualitatively different group from the other cancer types, and also suggest that the MSI-CIN- group is itself heterogeneous. PMID- 15531921 TI - Arsenic trioxide prevents radiation-enhanced tumor invasiveness and inhibits matrix metalloproteinase-9 through downregulation of nuclear factor kappaB. AB - Arsenic trioxide (ATO) has been implicated as a promising anticancer agent by inhibiting cell growth and inducing apoptosis in certain types of cancer cells. This study explored the antimetastasis property of arsenic, drew potential link between arsenic use and radiotherapy, and uncovered the specific mechanisms underlying these remarkable responses. Using gelatin invasion assay and intravasation assay, we report the novel finding that low-dose ATO (1 muM) reduced the intrinsic migration ability of HeLa cells and significantly inhibited radiation-promoted tumor invasive potential of CaSki cells without inducing apoptotic cell death. Using the murine Lewis lung carcinoma model, the control animals and ATO treatment animals (1 mg/kg i.p., twice weekly) displayed similar in vivo growth kinetics, whereas the radiation (30 Gy in one fraction) and concurrent treatment groups showed considerable growth inhibition. Importantly, although concurrent treatment did not enhance the effectiveness of radiation therapy to the primary tumor, further examination of the lungs revealed that all animals succumbed to radiation-accelerated lung metastases could be effectively treated by combination of ATO and radiation. Radiation-induced matrix metalloproteinase-9 (MMP-9) expression was significantly inhibited by ATO using sequential analysis of the expression of MMPs in xenografts. Supporting this observation, ATO directly downregulates radiation-induced MMP-9 mRNA expression by inhibiting nuclear factor kappaB activity in human cervical cancer cells. In sum, concurrent arsenic-radiation therapy not only achieves local tumor control but also inhibits distant metastasis. Experimental results of this study highlight a novel strategy in cancer treatment. PMID- 15531922 TI - Type I and type II reactions in TRAIL-induced apoptosis -- results from dose response studies. AB - Death receptor-induced apoptosis is paradigmatically mediated via the recruitment of FADD adapter molecule to the ligand/receptor complex and subsequent activation of caspase-8. However, several observations provided evidence that components of the mitochondrial apoptosis pathway are involved in death receptor-mediated apoptosis. In this regard, caspase-8-mediated activation of Bid induces the release of cytochrome c from the mitochondria, which, in turn, triggers the formation of the apoptosome protein complex, resulting in the activation of caspase-9. Whereas Bax or Bak were shown to be required for the proapoptotic effect of Bid, Bcl-2 was described to interfere with its action. Up to now, contradictory results regarding the role of Bcl-2 in TRAIL-induced apoptosis have been published. In order to study the influence of Bcl-2 on TRAIL-induced cell death more detailed, we utilized a tetracycline-regulated Bcl-2 expression system in Jurkat T cells. After having analysed the dose response for TRAIL-induced activation of caspase-8, -9, -3, breakdown of the mitochondrial membrane potential, and changes in the apoptotic morphology in cells expressing different Bcl-2 levels, we conclude that overexpression of Bcl-2 mediates a partial resistance towards lower doses of TRAIL that can be overcome when higher doses of TRAIL are applied. Thus, the requirement of the mitochondrial pathway for death receptor-induced apoptosis in type II cells should be reconsidered, since the protective effect of Bcl-2 is limited to lower TRAIL doses or early observation time points. PMID- 15531923 TI - Characterization of vinblastine-induced Bcl-xL and Bcl-2 phosphorylation: evidence for a novel protein kinase and a coordinated phosphorylation/dephosphorylation cycle associated with apoptosis induction. AB - Bcl-xL and Bcl-2 are phosphorylated in response to microtubule inhibitors, but the kinase(s) responsible and the functional significance have remained unclear. In this study, we investigated the characteristics of Bcl-xL and Bcl-2 phosphorylation in KB-3 carcinoma cells treated with vinblastine. In both asynchronous and synchronous cell cultures, Bcl-xL and Bcl-2 underwent a well defined and coordinated cycle of phosphorylation and dephosphorylation, with a lengthy period of phosphorylation preceding apoptosis induction, and with dephosphorylation closely correlated with initiation of apoptosis. Internally, validated inhibitors of JNK, ERK, p38(MAPK), or CDK1 failed to inhibit vinblastine-induced phosphorylation of Bcl-xL or Bcl-2. In vitro, Bcl-xL and Bcl 2 were poor substrates relative to c-Jun and ATF2 for active recombinant JNK1. Both Bcl-xL and Bcl-2 were localized primarily to the mitochondrial fraction in both control and vinblastine-treated cells, indicating that phosphorylation did not promote subcellular redistribution. Bcl-xL kinase activity was demonstrated in mitochondrial extracts from vinblastine-treated, but not control, cells. These findings suggest that phosphorylation of these key antiapoptotic proteins may be catalysed by a novel or unsuspected kinase that is activated or induced in response to microtubule damage. Furthermore, the same kinase and phosphatase system may be operating in tandem on both proteins, and phosphorylation appears to maintain their antiapoptotic function, whereas dephosphorylation may trigger apoptosis. These results provide evidence for a novel signaling pathway connecting microtubule damage to apoptosis induction, and help to clarify some of the controversy concerning the role of Bcl-2 phosphorylation in microtubule inhibitor-induced apoptosis. PMID- 15531924 TI - Mammary development and tumorigenesis in mice expressing a truncated human Notch4/Int3 intracellular domain (h-Int3sh). AB - Recently, we have identified a novel 1.8 kb human Notch4/Int3 RNA species (designated h-Int3sh). The h-Int3sh RNA encodes a protein that is missing the CBF1-binding region (RAM23) of the Notch 4/Int3 intracellular domain (ICD). Expression of h-Int3sh in the MCF10A 'normal' human mammary epithelial cell line has been previously shown to induce changes characteristic of oncogenic transformation, including anchorage-independent growth in soft agar. To study the consequences of h-Int3sh expression in vivo on mammary gland development and tumorigenesis, three transgenic mouse lines were established, in which the transgene is the Whey acidic protein (WAP) promoter linked to h-Int3sh. Expression of WAP-Int3sh was detectable in the mammary gland at day 15 of pregnancy in each transgenic line. Mammary gland development in all founder lines is normal and the females can lactate. WAP-h-Int3sh females from each of the founder lines develop mammary tumors, but with a long latency (average age of 18 months). Tumor development was associated with activation of Notch pathway, as evidenced by upregulation of Hes-1. The long latency of mammary tumors in WAP-h Int3sh mice could be due in part to the subcellular localization of h-Int3sh. Immunofluorescence analysis of transfected COS-1 cells showed that h-Int3sh is localized in the cytoplasm and nucleus, while Int3-ICD is detected only in the nucleus. We speculate that the Notch4/Int3 ICD-induced block to mammary gland development and tumorigenesis are consequences of an increasing gradient of CBF1 dependent Notch4/Int3 signaling. PMID- 15531925 TI - Enhanced growth of human met-expressing xenografts in a new strain of immunocompromised mice transgenic for human hepatocyte growth factor/scatter factor. AB - Downstream signaling that results from the interaction of hepatocyte growth factor/scatter factor (HGF/SF) with the receptor tyrosine kinase Met plays critical roles in tumor development, progression, and metastasis. This ligand receptor pair is an attractive target for new diagnostic and therapeutic agents, preclinical development of which requires suitable animal models. The growth of heterotopic and orthotopic Met-expressing human tumor xenografts in conventional strains of immunocompromised mice inadequately replicates the paracrine stimulation by human HGF/SF (hHGF/SF) that occurs in humans with cancer. We have therefore generated a mouse strain transgenic for hHGF/SF (designated hHGF-Tg) on a severe combined immunodeficiency (SCID) background. We report here that the presence of ectopically expressed hHGF/SF ligand significantly enhances growth of heterotopic subcutaneous xenografts derived from human Met-expressing cancer cells, including the lines SK-LMS-1 (human leiomyosarcoma), U118 (human glioblastoma), and DU145 (human prostate carcinoma), but not that of M14-Mel xenografts (human melanoma that expresses insignificant levels of Met). Our results indicate that ectopic hHGF/SF can specifically activate Met in human tumor xenografts. This new hHGF-Tg strain of mice should provide a powerful tool for evaluating drugs and diagnostic agents that target the various pathways influenced by Met activity. PMID- 15531926 TI - Roles of PDK-1 and PKN in regulating cell migration and cortical actin formation of PTEN-knockout cells. AB - Mutations in the tumor suppressor protein PTEN (phosphatase and tensin homologue deleted on chromosome 10) enhance cell migration, yet the underlying molecular mechanisms remain largely uncharacterized. Loss of PTEN in mouse embryonic fibroblasts (MEFs) correlates with striking cortical actin accumulation. However, how loss of PTEN leads to cortical actin formation and whether the presence of cortical actin contributes to the increased cell migration are unclear. Here we show that overexpression of dominant-negative forms of (DN) PTEN, RhoA or its kinase-dead (KD) effector, PKN, inhibited cortical actin formation, indicating that cortical actin of Pten(-/-) MEFs is mediated by the PTEN/Rho/PKN pathway. However, neither DN RhoA nor KD PKN inhibited the enhanced migration of Pten(-/-) cells, in contrast to the inhibitory effect of DN Rac. In agreement with the previous observation that DN Akt inhibits migration of Pten(-/-) cells, we demonstrate here that overexpression of KD PDK-1, the Akt kinase, reduces Pten(-/ ) cell migration. Furthermore, overexpression of DN forms of Akt, Rac, or PDK-1, all of which inhibit migration of Pten(-/-) cells, had no effect on cortical actin accumulation. Our findings suggest that PDK-1/Akt signaling pathway plays a major role in regulating cell migration induced by PTEN deficiency. PMID- 15531927 TI - BMP signals inhibit proliferation and in vivo tumor growth of androgen insensitive prostate carcinoma cells. AB - Prostate cancer is one of the most common cancers in men. Several lines of evidence have suggested that bone morphogenetic protein (BMP) signals play important roles in the generation and progression of prostate cancers. In the present study, we show that BMP-7 inhibits the proliferation of androgen insensitive PC-3 and DU-145 prostate cancer cells in a medium containing 1% fetal bovine serum, observed as decreased incorporation of [(3)H]thymidine and decreased cell number. Cell cycle analysis by flow cytometry showed an increased fraction of cells in the G1 phase and subsequent decrease in both S and G2/M phase after BMP-7 stimulation. BMP-7 caused an upregulation of the cyclin dependent kinase inhibitor (CDKI) p21(CIP1/WAF1), and decreased the activity of Cdk2, leading to hypophosphorylation of Rb proteins. Furthermore, in order to evaluate the impact of BMP signals on prostate tumor growth, we generated the PC 3 cell lines expressing a constitutively active BMP type I receptor (constitutively active (c.a.) activin receptor-like kinase (ALK)-6) in a tetracycline (Tet)-regulated manner. Tet/doxycycline-regulated expression of c.a.ALK-6 resulted in the inhibition of in vitro cell proliferation and reduction of the size of tumors derived from the PC-3 cells subcutaneously injected into immune-deficient mice. Collectively, these findings suggest that BMP signals inhibit growth and proliferation of prostate tumor cells through induction of CDKI. Furthermore, this is the first report of a role for BMP signaling in reducing growth kinetics of androgen-insensitive prostate tumors. PMID- 15531928 TI - Identification of PRC1 as the p53 target gene uncovers a novel function of p53 in the regulation of cytokinesis. AB - Our previous studies conducted in MCF7-ptsp53 cells have demonstrated that overexpression of the wild-type (wt) p53 at permissive temperature 32 degrees C leads to growth arrest at the G2/M phase of the cell cycle. To identify novel p53 regulated genes that are responsible for the p53-induced G2/M arrest, we conducted cDNA microarray analyses. The array results indicated that the mRNA level of protein regulator of cytokinesis (PRC1) was significantly decreased when the p53 transactivation activity was turned on, suggesting that PRC1 transcription could be downregulated by p53. In this study, we have extensively examined the functional role of p53 in the regulation of PRC1, a cell cycle protein that plays important roles during cytokinesis. We demonstrate that increased expression of the wt p53 either by exogenous transfection or chemical induction results in reduced mRNA and protein expression of PRC1 in HCT116 p53(+/+), HCT116 p53(-/-), MCF-7, T47D, and HeLa cells. Importantly, we show that the decreased PRC1 expression is accompanied by the appearance of binucleated cells, indicating the process of cell division after mitosis being inhibited. By isolation and characterization of a 3 kb genomic fragment containing the 5' flanking region and part of exon 1 of PRC1 gene, we demonstrate that p53 directly suppresses PRC1 gene transcription. We further locate the p53-responsive sequence to the proximal promoter region -214 to -163, relative to the transcriptional start site. The in vivo interaction of p53 with PRC1 gene promoter is further demonstrated by chromatin immunoprecipitation assay. Taken together, these new findings suggest that p53 may have important roles in the regulation of cytokinesis through controlling the transcription of PRC1. PMID- 15531933 TI - High-dose statin and COX-2 inhibitor therapy rapidly decreases C-reactive protein level in patients with unstable angina. AB - BACKGROUND AND AIM: Elevated levels of C-reactive protein (CRP) are associated with an increased risk of coronary events. The levels of CRP and other inflammatory markers are significantly elevated in patients with unstable angina. We hypothesised that a high-dose statin therapy alone or with cyclooxygenase-2 (COX-2) inhibitors, administered before coronary diagnostic or invasive procedures, can attenuate CRP elevation after the procedure and, consequently, more effectively reduce the rate of coronary events. METHODS: All patients with unstable angina in class III and IIB according to Braunwald classification were considered for inclusion in the present study. Finally, 60 patients with elevated CRP level (>3 mg/l) were randomised to three groups of pharmacological treatment before coronary angiography and subsequent angioplasty. Patients from group A received placebo, patients from group B - 80 mg of atorvastatin, and patients from group C - 80 mg of atorvastatin and 25 mg of rofecoxib. The levels of CRP were measured at baseline, after 3 days of therapy and 48 hours after invasive coronary procedure. RESULTS: The mean baseline CRP level in group A was 5.67+/ 2.82 mg/l, in group B - 4.7+/-1.32 mg/l, and in group C - 6.78+/-2.56 mg/l (NS). After three days of pharmacological treatment, the mean CRP level was 5.82+/-2.69 mg/l in group A (NS compared with baseline) and was significantly reduced in group B to 2.5+/-1.37 mg/l and in group C to 3.01+/-1.57 mg/l (p<0.0013 compared with group A). Measurements performed 48 hours after the procedure revealed a marked CRP level increase in group A (up to 24.54+/-5.48 mg/l) and a much lower increase in groups B and C (up to 3.02+/-2.0 mg/l and 7.31+/-2.96 mg/l, respectively). CONCLUSIONS: High-dose statin therapy alone or in combination with COX-2 inhibitor, administered before invasive coronary procedure in patients with unstable angina, rapidly lowers CRP levels. This therapy also reduces a marked CRP elevation typically occurring after invasive coronary intervention. Attenuation of inflammatory reaction may be crucial for the reduction of coronary events following invasive coronary interventions. PMID- 15531934 TI - Activation of generalised inflammatory reaction following electrical cardioversion. AB - BACKGROUND: Restoration of sinus rhythm in patients with atrial fibrillation (AF) is associated with an increased risk of thrombo-embolic complications due to delayed return of the left atrial and left atrial appendage systolic function. Direct current cardioversion (DC), used for AF termination, may cause myocardial injury and subsequent activation of inflammatory response. A C-reactive protein (CRP) is a non-specific marker of inflammation. AIM: To examine the effects of external DC of AF or atrial flutter (AFlut) on inflammatory processes. METHODS: The study group consisted of 35 patients (20 females and 15 males, mean age 67.9+/-9.7 years, range 46-83 years) with paroxysmal or persistent AF/AFlut who underwent elective DC. CRP plasma concentration was measured before and 24 hours after DC. RESULTS: The mean total DC energy was 431.2 J. CRP plasma concentration increased significantly following DC - from 3.9+/-3.4 ng/ml before DC to 7.2+/ 6.7 ng/ml after DC (p<0.0001). CRP level correlated with body mass index (r=0.34, p<0.05), however, this correlation became non-significant after inclusion of the presence of diabetes into the statistical model. There was also a positive correlation between CRP values before and after DC (r=0.72, p<0.0001). No correlation between CRP and gender, total power of DC nor the number of DC shocks was detected. CONCLUSIONS: External DC of AF/Aflut causes activation of inflammatory processes measured as a significant increase in the CRP plasma concentration. PMID- 15531935 TI - Primary coronary angioplasty in patients with ST segment elevation acute myocardial infarction and diabetes. AB - BACKGROUND: Subjects with diabetes constitute 13-25% of patients with ST segment elevation acute myocardial infarction (STEMI). In spite of the introduction of thrombolytic therapy, patients with STEMI and diabetes continue to have worse prognosis than those without diabetes. Primary percutaneous coronary intervention (PCI) has been shown in recent years to be the most effective therapy in patients with STEMI. AIM: To compare the outcome of STEMI patients with or without diabetes who underwent primary PCI. METHODS: The study group consisted of 500 consecutive patients with STEMI. The occurrence of major adverse cardiac events (MACE) which included death, reinfarction or repeated PCI of the target vessel, was analysed peri-operatively and during a six-month follow-up period.Results. Diabetes was diagnosed in 68 (13.6%) patients. The mean time duration from the onset of STEMI symptoms to treatment was similar in patients with or without diabetes (230+/-97 min vs 231+/-139 min, NS). Patients with diabetes were older (61.9+/-8.9 vs 57.9+/-10.8 years, p=0.004), had higher body mass index (29+/-4 vs 27+/-5, p=0.002), more frequent history of coronary artery disease (57.4% vs 37.9%, p=0.002), higher prevalence of arterial hypertension (71.6% vs 56.8%, p=0.02) and more frequently the left anterior descending artery as the infarct related artery (58.8% vs 42.1%, p=0.01). Immediately after PCI, epicardial and myocardial reperfusion rates were lower in patients with rather than without diabetes (TIMI 3: 84.9% vs 91.3%, p=NS, cTFC: 32+/-26 vs 22+/-16, p<0.0001, and MPG3: 25% vs 41.9% p=0.008). Diabetes increased the risk of MACE during in hospital period by 2.7 times. The rate of MACE during a six-month follow-up period was almost two times higher in patients with rather than without diabetes (death: 8.8% vs 5.1%, reinfarction: 1.5% vs 1.2%, repeated PCI: 11.8% vs 6.9%). CONCLUSIONS: Primary PCI-achieved epicardial and myocardial reperfusion rate is lower in STEMI patients with rather than without diabetes. The presence of diabetes almost doubles the risk of MACE during a six-month follow-up. PMID- 15531936 TI - Is the assessment of lipid profile performed up to seventy two hours from hospital admission due to acute coronary syndrome still valid? AB - BACKGROUND: Serum concentration of lipids and lipoproteins changes during the course of acute coronary syndrome (ACS). Total cholesterol and LDL-cholesterol levels decrease. Also HDL-cholesterol and triglyceride levels are not stable. Therefore, it has been suggested that reliable assessment of lipid profile can be performed either during the first 24 hours or after 6 weeks or even 3 months from the onset of ACS. AIM: To examine whether lipid parameters remain stable during the first 72 hours of hospitalisation due to ACS. METHODS: The study group consisted of 84 patients, aged 30-88 years, hospitalised due to ACS, of whom 90% underwent primary percutaneous coronary intervention (PCI). In all patients lipid profile was assessed twice. The first measurement was performed as soon as possible, within 24 hours from hospital admission. The second measurement was obtained within 24 hours from the first sample (group I, n=42), between 24 and 48 hours (group II, n=28), or between 48 and 72 hours (group III, n=15) from the first measurement. Lipid parameters obtained from the first measurement were compared with those obtained during the second assessment. RESULTS: There were no significant differences between lipid parameters measured during the first and second measurement within all three investigated groups nor were there any significant inter-group differences. CONCLUSIONS: Lipid profile assessed within 72 hours from the hospital admission due to ACS is as valid as the measurement performed within the first 24 hours. PMID- 15531937 TI - Primary direct stenting versus endoscopic atraumatic coronary artery bypass surgery in patients with proximal stenosis of the left anterior descending coronary artery--a prospective, randomised study. AB - BACKGROUND: The dynamic development of interventional cardiology resulted in an increasing proportion of patients treated with various forms of coronary angioplasty instead of surgery. On the other hand, it has been well established that the results of coronary artery by-pass surgery of the left anterior descending (LAD) coronary artery with the use of the internal mammary artery are excellent. AIM: To compare the results of primary direct stenting (PDS) and endoscopic atraumatic coronary artery bypass (EACAB) surgery in patients with an isolated proximal LAD type A or B1 lesion. METHODS: This prospective and randomised study included 100 patients with an isolated critical (> or =70%) LAD stenosis who underwent PDS (n=50) or EACAB (n=50). RESULTS: After a six-month follow-up period, 32 (64%) PDS patients and 47 (94%) EACAB patients were angina free. The rate of major cardiac adverse events (MACE) was significantly higher in the PDS group than in surgically treated patients (p<0.05). After one year of follow-up, 40 (80%) PDS-treated patients and all 50 EACAB patients had no recurrences of angina. After two-year follow-period, the survival rate without MACE was significantly higher in the EACAB group than in the PDS-treated patients (94% vs 76%, p<0.05). CONCLUSIONS: Minimally invasive cardiac surgery is an alternative method to direct stenting in the treatment of patients with proximal LAD stenosis. PMID- 15531938 TI - [Successful angioplasty of the left anterior descending artery in a woman with Takayasu arteritis and associated coronary disease]. AB - Takayasu arteritis is a rare, inflammatory disease affecting mainly young women and is more prevalent in Asia and Latin America. Its etiology is obscure. It involves the aorta and its main branches. The typical lesions are represented by obliteration and aneurysmatic dilatation of arteries and ostial stenosis. We report a case of a 51-year-old woman with Takayasu arteritis and coronary disease, complaining of angina pectoris CCS class IV. The patient underwent successful angioplasty of LAD. During 6-month follow-up she remained asymptomatic. Various therapeutic options of revascularization are reviewed. PMID- 15531940 TI - [Univentricular heart, common atrium, single atrio-ventricular valve--is it possible in humans?]. AB - Two extremely rare cases of adults with univentricular hearts of right ventricular morphology are presented. Pathogenesis and pathophysiology of univentricular circulation as well as anatomic variants, short information on prognosis and management are briefly discussed. Echocardiographic diagnosis of univentricular heart is described. The first case was a 23-year-old housewife with univentricular heart, functionally common atrium, single atrio-ventricular valve of tricuspid morphology, transposition of great arteries and significant subvalvular pulmonary stenosis. The second case was a 19-year-old student with univentricular heart with moderately depressed systolic function, common atrium, single atrio-ventricular valve of tricuspid morphology, mitral atresia, and non communication residual left ventricle. Both patients led a normal life with little to moderate limitations of everyday activity. PMID- 15531939 TI - [Recurrent ventricular fibrillation during a febrile illness as the first manifestation of Brugada syndrome--a case report]. AB - A case of a 33-year-old male who was admitted to the hospital due to recurrent ventricular fibrillation during a febrile illness is presented. Initially, the patient was diagnosed with acute myocardial infarction and received thrombolytic treatment. Echocardiography and coronary angiography were normal. Right precordial ECG leads recorded one and two intercostal spaces higher than normal as well as ECG obtained following ajmaline administration revealed a typical Brugada pattern. PMID- 15531941 TI - [Biochemical markers of cardiac injury and strain in the risk stratification of patients with acute pulmonary embolism]. PMID- 15531943 TI - [Electrocardiogram of the month: Syncope]. PMID- 15531944 TI - [Congenital malformation of the anterior mitral leaflet in a neonate--a case report]. PMID- 15531945 TI - [Angiogram of the month: Myocardial ischemia]. PMID- 15531946 TI - [Cardiovascular complications of oncological therapy]. PMID- 15531949 TI - Haematopoietic graft engineering. AB - Although haematopoietic stem cell transplantation (HSCT) is a powerful treatment modality, it is a blunt instrument against cancer and diseases of the haematopoietic and immune system. Various techniques have been developed to engineer the haematopoietic stem cell (HSC) graft to enable it to perform its task more effectively. These techniques include the removal of cells which cause graft-versus-host disease (GVHD), the eradication of cells which might cause relapse, the expansion of donor cells when there is an inadequate cell dose, and the addition of selected cells to improve graft function. In this review, we will discuss each of these means of haematopoietic graft manipulation in turn and then touch on some regulatory requirements in the field of haematopoietic graft engineering. While the science is still prepubescent, it has passed its infancy and further developments in the next decade or so should allow it to be taken to a wider scale to benefit more patients. PMID- 15531950 TI - Unrelated umbilical cord blood transplantation in children and adults. AB - Umbilical cord blood (UCB) has recently been explored as an alternative haematopoietic stem cell (HSC) source for allogeneic immunotherapy in both adults and paediatric patients with haematological malignancies and marrow failure syndromes. The relative ease of procurement, tolerance of 1-2 antigen human leukocyte antigen (HLA) mismatch and the lower than anticipated risk of severe graft-versus-host disease has made UCB an appealing alternative to marrow-derived HSC. Results from various registries and institutions observed graft cell dose to be the major factor determining engraftment and survival in unrelated UCB transplant recipients. Given that adults are larger than children, there was still limited enthusiasm for the use of UCB in adults. The use of reduced intensity or nonmyeloablative preparative regimens to allow engraftment of UCB broadens the scope of patients who may benefit from allogeneic immunotherapy, particularly the elderly and medically infirm patients with no matched sibling donor. Further studies on improving graft cell dose such as the use of ex vivo expansion of UCB cells and multiple-unit transplant are currently being pursued, so as to make this potentially curative procedure available to more patients. PMID- 15531951 TI - In vitro biological characteristics of human cord blood-derived megakaryocytes. AB - INTRODUCTION: Umbilical cord blood (CB) has been used as an alternative source for haematopoietic stem cell transplantation (HSCT) in recent years. However, delayed platelet recovery is frequently associated with CB HSCT. Megakaryocytes (Mk) are the specialised precursors of platelets and they are among the rarest haemopoietic cell types. Despite the rapid expansion of our knowledge of megakaryopoiesis in recent years, many questions, such as the molecular regulatory mechanisms in Mk differentiation and maturation, platelet formation and release, remain unanswered in CB-derived megakaryopoiesis. Variations can be seen from the literature by individual investigators using different approaches for Mk-specific differentiation and maturation induction. The development of in vitro culture methods to obtain sufficient numbers of Mks from readily available haematopoietic stem cells is of value for both basic research and clinical applications. MATERIALS AND METHODS: The CD34+ cells from cord blood samples were cultured in serum-free medium with haematopoietic growth factors (GFs), such as IL-3, stem cell factor (SCF), and thrombopoietin (Tpo). The differentiation of Mk was monitored using Mk- and platelet-specific monoclonal antibodies and flow cytometric analysis. The morphology of the cultured cells was studied by both light and electronic microscopy (LM and EM). The involvement of the human Notch gene family members was studied by real time-polymerase chain reaction (RT-PCR). Maturation of the cultured Mks was studied using flow cytometric analysis for both platelet-specific surface markers and enodomitosis. Platelet activation was assessed in the cytoplasmic fragments harvested from the cultures. RESULTS: Specific Mk differentiation of >70% resulted from a 2-step culture approach using IL-3, SCF and Tpo for 7 days followed by Tpo only for another 14 days. RT-PCR showed high-level expression of both Notch-1 and its ligand, Jagged-1, in the cultured Mks. Limited levels of polyploidy (>4N, endomitosis, EnM) were observed in the cultured Mks. The results also showed that the cytoplasmic fragments from the cultures responded to platelet activation reagents, including ADP and collagen, marked by upregulation of platelet-specific activation markers, such as CD62P (P-selectin) and PAC-1 (gpalphaIIbbeta3). CONCLUSION: The methods used in this study are specific for differentiation of Mk from CB CD34+ cell, which can partially mature and produce functional platelets in vitro. This approach for human Mk differentiation could be further optimised and may be adapted on larger scales for clinical purposes. PMID- 15531952 TI - Ocular surface stem cells and disease: current concepts and clinical applications. AB - Corneal and conjunctival epithelial stem cells are responsible for the homeostasis and regeneration of the ocular surface epithelium. Corneal epithelial stem cells reside in the basal region of the limbus, while the conjunctival forniceal region appears to be the site that is enriched in conjunctival stem cells. Ocular surface disease arising from limbal stem cell deficiency is characterised by persistent epithelial defects, corneal vascularisation, chronic inflammation, scarring and conjunctivalisation, resulting in visual loss. Limbal stem cell transplantation replaces the corneal stem cell population in these eyes with the hope of restoring vision. More recently, the use of bioengineered ocular surface tissue-equivalents has had promising results, and may represent the future for replacement and regeneration of ocular tissues in various ocular disorders. PMID- 15531953 TI - Axoglial interaction via the notch receptor in oligodendrocyte differentiation. AB - INTRODUCTION: Increasing evidence has revealed that the Notch signalling pathway is one of the pivotal systems that mediate oligodendrocyte development. The Notch receptor is a type I transmembrane molecule that represents a novel cellular signalling paradigm, namely, regulated intramembrane proteolysis (RIP). METHOD: The typical Notch ligands, such as Delta, Serrate/Jagged and Lag2 (DSL), promote the formation of oligodendocyte precursor cells (OPCs) and maintain them in an uncommitted stage, thus retarding oligodendrocyte appearance in the central nervous system (CNS). RESULTS: In contrast, our recent studies have revealed that F3/contactin, a GPI-linked neural adhesion molecule, interacts with Notch and speeds up the generation and maturation of oligodendrocytes. CONCLUSIONS: Considering the distinct, albeit somewhat overlapping expression patterns of F3 and DSL in the CNS, the Notch receptor appears to function ligand-dependently during oligodendrocyte development. This multipotentiality may well designate the Notch receptor as one of the therapeutic targets that one can manoeuvre to treat demyelinating diseases, such as multiple sclerosis, that is characterised by chronic myelin degeneration. PMID- 15531954 TI - Clinico-pathological analysis of myelodysplastic syndromes according to French American-British classification and international prognostic scoring system. AB - INTRODUCTION: The aim of this study was to analyse the clinico-pathological features of a cohort of patients with myelodysplastic syndromes (MDS). MATERIALS AND METHODS: The clinical and pathological data of 43 MDS patients over a 3-year period were reviewed. Survival analysis was performed according to the French American-British (FAB) classification and International Prognostic Scoring System (IPSS) using the Kaplan-Meier method. Selected published studies for comparison were identified from MEDLINE search. RESULTS: The patients were followed up for a median duration of 175 days (range, 2 to 1044 days). The median survival for refractory anaemia (RA) and refractory anaemia with ringed sideroblasts (RARS) has not been reached, but that for refractory anaemia with excess blasts (RAEB), refractory anaemia with excess blasts in transformation (RAEB-T) and chronic myelomonocytic leukaemia (CMML) was 250 days, 49 days and 44 days, respectively. The median survival for the low-risk and intermediate-1 IPSS categories has not been reached, while that for the intermediate-2 and high-risk categories was 58 days and 49 days, respectively. The survival analyses, according to the FAB classification and IPSS system, were statistically significant (P <0.05). Comparison of our data with those from neighbouring and Western countries revealed both similarity and disparity. We also noted different cytogenetic information in our cohort of patients. CONCLUSIONS: We found distinctly unique cytogenetic and clinico-pathological characteristics in our MDS patients. However, whether true biological differences exist among MDS patients in different geographies and populations with different genetic and environmental backgrounds require further large multinational study. PMID- 15531955 TI - Serum interleukin-2, interleukin-6, tumour necrosis factor-alpha and nitric oxide levels in patients with Behcet's disease. AB - INTRODUCTION: Behcet's disease (BD) is a chronic systemic disorder characterised by oral and genital ulcerative lesions, ocular and cutaneous manifestations. Cytokines are the major mediators of immunologic and inflammatory reactions. Nitric oxide is reactive nitrogen intermediate which plays a key role in pathogenesis of many inflammatory and autoimmune skin diseases. The study was conducted to determine serum interleukin-2 (IL-2), interleukin-6 (IL-6), tumour necrosis factor (TNF)-alpha and nitric oxide levels in relation to the pathogenesis of Behcet's disease. MATERIALS AND METHODS: Serum IL-2, IL-6, and TNF-alpha levels were measured with test kits by enzyme-linked immunosorbent assay (ELISA) method, while serum nitric oxide levels were determined with a test kit by colorimetric method. Serum IL-2, IL-6, TNF-alpha and nitric oxide concentrations in 27 patients with Behcet's disease and in 16 healthy controls were determined by extrapolation from their standard curves. The significance of the mean differences between the 2 groups was assessed by the Mann-Whitney U test. RESULTS: The serum levels of IL-2, IL-6, TNF-alpha, and nitric oxide concentrations in patients with BD were significantly higher than those of the controls (P <0.001). CONCLUSION: Our results suggest that elevated levels of IL 2, IL-6, TNF-alpha, and nitric oxide in Behcet's disease appear to be related to the disease. PMID- 15531956 TI - Breast ultrasound in women with familial risk of breast cancer. AB - INTRODUCTION: The aim of this study was to assess the performance and value of breast ultrasound in women with familial risk of breast cancer. MATERIALS AND METHODS: From an initial dataset of 245 women with positive family history who had breast cancer surveillance utilising mammography or magnetic resonance imaging (MRI) between November 1994 and February 2001, 179 subjects with follow up data were selected. Eighty-four women had breast ultrasound done with histopathological correlation available from 48 breast biopsies performed in 42 women. RESULTS: The sensitivity of ultrasound, mammography and MRI was 83.3%, 53.9% and 93.3%, respectively. The specificity of ultrasound, mammography and MRI was 65.5%, 85.7% and 63.6%, respectively. Ultrasound was the imaging modality with intermediate sensitivity, specificity, negative predictive value (NPV) and cancer detection rate. The sensitivity, specificity, positive predictive value (PPV), NPV and accuracy of combined mammography and ultrasound were 92.9%, 62.5%, 52.0%, 95.2% and 71.7%, respectively. These results did not differ significantly from MRI. Almost two-thirds of the breast biopsies were performed under ultrasound guidance. CONCLUSIONS: Although breast ultrasound screening per se was not assessed in this study, extrapolation of these results to sonographic screening of high familial risk women would come at a better specificity to MRI, albeit with a 10% decrease in sensitivity but at a fraction of the cost of MRI. Ultrasound also provides the advantage of convenient imaging guidance for biopsy. Employing ultrasound following mammography would match MRI in sensitivity, specificity, PPV, NPV and accuracy, and should not be ignored in these women. PMID- 15531957 TI - Audit of diagnostic and interventional craniocervical catheter angiographic procedures at the Singapore General Hospital. AB - INTRODUCTION: Catheter angiography is an established imaging modality of evaluating cerebral and head and neck vascular diseases. It is, however, an invasive procedure with a small risk of complications. The aim of our study was to evaluate the prevalence of peri-procedural complications in a local hospital setting. MATERIALS AND METHODS: A total of 88 patients underwent diagnostic and interventional craniocervical procedures over 6 months in our department. The casenotes of 83 patients were retrospectively reviewed for complications arising from a total of 99 procedures carried out. RESULTS: A new focal neurological deficit developed in 3 different patients after a procedure, giving a prevalence of 3.0%. All these occurred in diagnostic procedures and were permanent deficits with correlative computed tomography (CT) or magnetic resonance (MR) imaging findings of acute cerebral infarction. All these occurred in high-risk patients who had severe underlying cerebrocarotid vascular compromise. There was 1 case of contrast medium-induced nephropathy (1.0%), occurring in a patient with pre existing renal impairment. Local complications included 1 case of iatrogenic external iliac artery dissection (1.0%) and 5 cases (5.1%) of small and uncomplicated puncture site groin haematomas. CONCLUSION: The most significant complication associated with a craniocervical angiographic procedure was the development of post-procedural stroke in patients with significant preexisting cerebrocarotid vascular compromise. In the absence of this risk factor, craniocervical catheter angiography is a relatively safe procedure. PMID- 15531958 TI - Improving the diagnosis related grouping model's ability to explain length of stay of elderly medical inpatients by incorporating function-linked variables. AB - INTRODUCTION: This study first aimed to determine the adequacy of the Diagnosis Related Grouping (DRG) model's ability to explain (1) the variance in the actual length of stay (LOS) of elderly medical inpatients and (2) the LOS difference in the same cohort between the departments of Geriatric Medicine (GRM) and General Medicine (GM). We then looked at how these explanatory abilities of the DRG changed when patients' function-linked variables (ignored by DRG) were incorporated into the model. MATERIALS AND METHODS: Basic demographic data of a consecutively hospitalised cohort of elderly medical inpatients from GRM and GM, as well as their actual LOS, discharge DRG codes [with their corresponding trimmed average length of stay (ALOS)] and selected function-linked variables (including premorbid functional status, change in functional profile during hospitalisation and number of therapists seen) were recorded. Beginning with ALOS, function-linked variables that were significantly associated with LOS were then added into two multiple liner regression models so as to quantify how the functional dimension improved the DRGs' abilities to explain LOS variances and interdepartmental LOS differences. Forward selection procedure was employed to determine the final models. For the interdepartmental analysis, the study sample was restricted to patients who shared common DRG codes. RESULTS: 114 GRM and 118 GM patients were studied. Trimmed ALOS alone explained 8% of the actual LOS variance. With the addition of function-linked variables, the adjusted R2 of the final model increased to 28%. Due to common code restrictions, the data of 79 GRM and 78 GM patients were available for the analysis of interdepartmental LOS differences. At the unadjusted stage, the median stay of GRM patients was 4.3 days longer than GM's and with adjustments made for the DRGs, this difference was reduced to 3.9 days. Additionally adjusting for the patients' functional features diminished the interdepartmental LOS discrepancy even further, to 2.1 days. CONCLUSION: This study demonstrates that for elderly medical inpatients, the incorporation of patients' functional status significantly improves the DRG model's ability to predict the patients' actual LOS as well as to explain interdepartmental LOS differences between GRM and GM. PMID- 15531959 TI - A study on SARS awareness and health-seeking behaviour - findings from a sampled population attending National Healthcare Group Polyclinics. AB - INTRODUCTION: The study aimed to assess the effectiveness of massive SARS public education effort on SARS awareness and the conduct of those suspected of having SARS. MATERIALS AND METHODS: Five hundred and ninety-three respondents attending the National Healthcare Group Polyclinics (NHGP) participated in the survey from 9 to 13 June 2003. Associations between awareness of SARS symptoms and (i) first action to be taken and (ii) mode of transportation used, if the respondent was suspected of having SARS, were analysed using Chi-square or Fisher's exact tests. Logistic regression was performed to adjust for relevant covariates. RESULTS: The majority (92.7%) of the respondents were aware of SARS symptoms. Television (91.6%), newspaper (65.2%) and radio (30.4%) formed the top 3 sources of information on SARS. Slightly more than half (51.6%) of those who suspect themselves of having SARS would choose to visit their primary health care doctors, while 22.7% of the respondents would go to Tan Tock Seng Hospital (TTSH). If they suspected themselves to have SARS, most (84.9%) of the 578 respondents would react appropriately by taking the SARS ambulance or driving themselves to TTSH. However, 60 respondents would nonetheless take public transport to TTSH [by taxi 8.5%, mass rapid transit (MRT) or bus 1.9%]. In particular, the retired with lower educational levels were likely to be oblivious both to the symptoms of SARS and the possible consequences of travelling by inappropriate transport. CONCLUSION: Despite more than 2 months of intensive SARS public education in Singapore, there remain important gaps in knowledge and appropriate behaviour that have to be bridged. PMID- 15531960 TI - A risk reduction approach for schizophrenia: the Early Psychosis Intervention Programme. AB - Schizophrenia is a serious mental disorder with a profound impact on patients, their caregivers and society. Most patients who develop schizophrenia experience a prodromal phase where there is a change in premorbid functioning prior to the onset of frank psychotic symptoms. Once psychosis sets in, the subsequent course can be variable but schizophrenia tends to run a chronic course, resulting in considerable disabilities. The prognosis of schizophrenia could potentially be improved by reducing the duration of untreated psychosis (DUP). The Early Psychosis Intervention Programme (EPIP) in Singapore adopts a risk-reduction approach. It seeks to reduce the DUP through public education, networking with the primary healthcare providers (general practitioners, counsellors, traditional healers), and the screening of conscripts into the Singapore Armed Forces. Integral to this programme is a service for those in the prodromal phase of psychosis, that addresses the concerns of stigmatisation and pharmacotherapeutic interventions. Our tertiary prevention strategies aim to reduce mortality and morbidity, and to improve the quality of the lives of the individuals diagnosed with this disorder through a comprehensive and holistic management programme that comprises case-management, the judicious use of antipsychotics, and various psychosocial interventions. Since EPIP's initiation in April 2001, there has been a steady increase in the number of individuals screened and accepted into the programme. Our networking strategy is gaining momentum and there has been a significant increase in the number of primary healthcare providers whom we have trained to identify early signs of psychosis. However, there remain various challenges which we are yet to overcome. PMID- 15531962 TI - Initial experience of macular translocation in Singapore - one-year results. AB - INTRODUCTION: This paper reports the 1-year results of the first 2 cases of macular translocation in Singapore. CLINICAL PICTURE: A 66-year-old female and a 45-year-old male Chinese presented with subfoveal choroidal neovascularisation (CNV) in their right eyes. The woman's condition was secondary to pathological myopia while the man's was idiopathic. Their preoperative best-corrected visual acuities were 6/15-2 and 6/30, respectively. TREATMENT: Both patients underwent macular translocation with punctate retinotomies and chorioscleral infolding (limited macular translocation) in their affected eye. OUTCOME: Both patients achieved effective macular translocation postoperatively. Their CNVs became extrafoveal and were ablated with conventional laser photocoagulation in the early postoperative period. They did not recur and their visual acuities improved to 6/9-1 and 6/12 at 1 year postoperatively, respectively. CONCLUSION: Macular translocation is a new treatment modality that offers patients with subfoveal CNV a chance of improving their vision, potentially to a level that may allow reading and driving. PMID- 15531961 TI - Imaging findings in acute neck infection due to pyriform sinus fistula. AB - INTRODUCTION: Pyriform sinus fistula is a congenital branchial pouch abnormality that is often overlooked as a cause of acute neck infection in children. Our aim is to demonstrate the value of various imaging modalities (ultrasound, computed tomography [CT], barium oesophagraphy) in its diagnosis. MATERIALS AND METHODS: The preoperative imaging findings of 5 patients with surgically proven pyriform sinus fistula who presented with acute neck infection between September 2001 and March 2003 were retrospectively reviewed. CT was performed in all patients, 4 patients had barium oesophagraphy and 3 had an ultrasound scan. RESULTS: All 5 patients suffered from upper respiratory tract infection within a week of developing a tender swelling on the left side of the neck. Four patients had a history of recurrent neck infections. CT depicted inflammation of the left perithyroid soft tissue and adjacent left thyroid lobe in every case. In 2 cases, CT demonstrated the presence of a pyriform sinus fistula. Ultrasound, performed in 3 patients, correlated strongly with the CT findings. It also showed gas within a fistula in 1 case. Barium oesophagraphy clearly delineated the fistula in 3 out of 4 cases. CONCLUSION: Ultrasound and CT accurately showed the presence of acute neck infection and could demonstrate the pyriform sinus fistula. Barium oesophagraphy most clearly depicted the presence and course of the fistula. Recurrent left-sided neck infection in a child should alert the physician to the possibility of an underlying pyriform sinus fistula and imaging should be performed to confirm its presence. PMID- 15531963 TI - A case of a diaphragmatic rupture complicated with lacerations of stomach and spleen caused by a violent cough presenting with mediastinal shift. AB - INTRODUCTION: Diapraghmatic rupture is a clinical case that is mostly seen following a blunt thoracoabdominal trauma or is rarely reported as spontaneously induced by various factors. CLINICAL PICTURE: A 28-year-old man presented as an emergency with shortness of breath and severe abdominal pain following a violent cough. His chest radiography and computed tomography demonstrated left diaphragmatic rupture, mediastinal shift and herniation of gastric fundus into the pleural cavity. TREATMENT: Left thoracotomy for the replacement of herniated gastric fundus and median laparotomy for the repair of serosal layer of gastric fundus and a diaphragmatic gap were performed. OUTCOME: He made an uneventful recovery. CONCLUSIONS: Diaphragmatic ruptures may be caused by violent coughing with serious life-threatening complications. PMID- 15531964 TI - Case reports of two biopsy-proven patients with Creutzfeldt-Jakob disease in Singapore. AB - INTRODUCTION: Creutzfeldt-Jakob disease (CJD) is the most common transmissible human subacute spongiform encephalopathy. There is limited literature on CJD in Southeast Asia. We describe the clinical course and diagnostic evaluation of 2 Singapore patients with biopsy-proven CJD. CLINICAL PICTURE: Two patients presented with non-specific symptoms such as withdrawal, forgetfulness, asthenia, giddiness and insomnia. Both patients had spontaneous myoclonic jerks and impairment of multiple neurologic systems (visual, pyramidal, cerebellar and neurocognitive systems). Magnetic resonance imaging and electroencephalography provided helpful supportive evidence. Diagnosis of CJD was established on brain biopsy. Histological features included spongioform degeneration, neuronal cell loss and astrocytosis. TREATMENT/OUTCOME: Treatment remains palliative. Deterioration in their clinical condition was relentless, progressing to a totally dependent state within 10 to 12 months. CONCLUSION: The early features of CJD can be varied and non-specific. It is important for physicians from different specialties to be cognisant of the clinical manifestations of CJD and the appearance of supportive and definitive investigations. PMID- 15531965 TI - Acute isolated sphenoid sinusitis. AB - INTRODUCTION: Acute isolated sphenoid sinusitis is seen in fewer than 3% of all cases of sinusitis. It is frequently misdiagnosed because of its vague symptoms and the paucity of clinical findings. We report 2 cases of isolated acute isolated sphenoid sinusitis with unusual presentations. CLINICAL PICTURE: Both patients presented with acute headache, eye pain and fever, and were provisionally diagnosed as meningitis. In 1 case, the symptoms were on the contralateral side of the sphenoid infection. Intracranial complications were also present. TREATMENT: Treatment included intravenous antibiotics and endoscopic sphenoidotomy. OUTCOME: Both patients recovered with no residual neurological disability. CONCLUSION: Acute sphenoiditis usually presents with subtle symptoms and elusive physical findings and hence a high index of suspicion is necessary. Complications may arise due to the close proximity of important structures to the sphenoid sinus. Uncomplicated cases can resolve with optimal antibiotic therapy if diagnosed and treated early. Persistence or progression of disease with development of intracranial complications are indications for immediate surgical drainage. PMID- 15531966 TI - Objective analysis of gastroesophageal reflux after laparoscopic heller myotomy: an anti-reflux procedure is required. AB - BACKGROUND: Controversy exists over the necessity of performing a concurrent antireflux procedure with a Heller myotomy. We therefore sought to objectively analyze gastroesophageal reflux following laparoscopic Heller myotomy where an antireflux procedure was not performed. METHODS: A prospective database of 66 cases of laparoscopic Heller myotomy performed between November 1996 and June 2002 was reviewed. Previous, concurrent, or subsequent fundoplication was performed in 12 patients; therefore 54 patients without antireflux procedures were available for analysis. Follow-up included symptomatic assessment in 50 patients (93%). Heartburn was assessed on a four-point scale with clinical significance defined as >2 episodes/week. Objective testing, including endoscopy, esophagogram, manometry, and 24-h pH monitoring, was offered to all patients. Objective evidence of reflux was defined as the composite endpoint of positive 24 h pH monitoring or esophagitis on endoscopy. RESULTS: Significant heartburn was reported in 15 of 50 patients (30%). Positive 24-h pH recordings were seen in 11 of 22 patients tested while esophagitis was seen in 13 of 21 patients tested, resulting in objective evidence of reflux in 18 of 30 patients tested (60%). Of these 18 patients, seven did not have significant heartburn. All 12 patients without objective reflux did not have significant heartburn. Therefore, of the 30 patients with objective testing, seven (23%) had objective reflux without subjective heartburn (silent reflux). CONCLUSION: Objective analysis reveals an unacceptable rate of gastroesophageal reflux in laparoscopic Heller myotomy without an antireflux procedure. We therefore recommend performing a concurrent antireflux procedure. PMID- 15531967 TI - Open surgery induces a dramatic decrease in circulating intact IGFBP-3 in patients with colorectal cancer not seen with laparoscopic surgery. AB - BACKGROUND: As shown earlier by the authors via Western blot analysis, open (OS) but not laparoscopic surgery (LS) induces a qualitative decrease in plasma insulin-like growth factor-binding protein 3 (IGFBP-3) levels on postoperative day 1 (POD 1). Intact IGFBP-3 has tumor suppressive effects, but its degradation products do not. Enzyme linked immunoassay (ELISA) inevitably measures both. In this study, using a novel combined Western blot and ELISA analysis method, precise plasma levels of intact IGFBP-3 on POD2 after open and closed colorectal cancer resection (stage I-III) were determined. METHODS: This study included 15 OS patients with a mean incision length of 26.7 +/- 15.5 cm and 16 LS patients with a mean incision length of 5.3 +/- 3.1 cm. Intact IGFBP-3 levels were determined via ELISA and Western blot analysis in plasma collected preoperatively and postoperatively. RESULTS: In the OS patients, the mean preoperative concentration of intact 43-45 kDa IGFBP-3 protein was 1920 +/- 1430 ng/ml. It decreased dramatically on POD2 to 355 +/- 545 ng/ml (p < 0.005). In the LS group, no significant difference was noted between the preoperative level (1305 +/- 807 ng/ml) and the POD2 level (922 + 714 ng/ml). CONCLUSIONS: Open cancer resection, unlike its minimally invasive alternative, induces a dramatic decrease in concentration of intact IGFBP-3, which may have important implications with regard to colon cancer recurrence. PMID- 15531968 TI - Should laparoscopic paraesophageal hernia repair be abandoned in favor of the open approach? AB - BACKGROUND: The most appropriate approach to the repair of large paraesophageal hernias remains controversial. Despite early results of excellent outcomes after laparoscopic repair, recent reports of high recurrence require that this approach be reevaluated. METHODS: For this study, 60 primary paraesophageal hernias consecutively repaired at one institution from 1990 to 2002 were reviewed. These 25 open transabdominal and 35 laparoscopic repairs were compared for operative, short-, and long-term outcomes on the basis of quality-of -life questionnaires and radiographs. RESULTS: No difference in patient characteristics was detected. Laparoscopic repair resulted in lower blood loss, fewer intraoperative complications, and a shorter length of hospital stay. No difference in general or disease-specific quality-of-life was documented. Radiographic follow-up was available for 78% open and 91% laparoscopic repairs, showing anatomic recurrence rates of 44% and 23%, respectively (p = 0.11). CONCLUSIONS: Laparoscopic repair should remain in the forefront for the management of paraesophageal hernias. However, there is considerable room for improvement in reducing the incidence of recurrence. PMID- 15531969 TI - Laparoscopic palliation of polycystic liver disease. AB - The role of laparoscopic surgery in the management of polycystic liver disease (PCLD) is not well defined. The authors hypothesized that laparoscopic fenestration for PCLD relieves symptoms caused by polycystic liver disease. In this study, 11 patients underwent 20 laparoscopic cyst fenestration operations as treatment for symptoms of their PCLD. Symptoms leading to surgery were pain and pressure in 15 (75%) and early satiety in 12 (60%) patients. The median hospital stay was 1 day. The symptoms resolved postoperatively in all the patients. An additional laparoscopic fenestration was required in six (55%) patients for recurrent symptoms. The average time to reoperation was 22 +/- 16 months. Two patients required hepatic transplantation. Initial symptom resolution occurred in all the patients undergoing redo fenestration. The authors conclude that laparoscopic fenestration for PCLD is safe, results in minimal "down" time and relieves the symptoms caused by PCLD. Symptomatic relief usually is temporary, and repeat surgery is required for recurring symptoms in half of the patients. PMID- 15531970 TI - Three port vs standard laparoscopic cholecystectomy. PMID- 15531971 TI - Validation of a six-task simulation model in minimally invasive surgery. AB - BACKGROUND: The content validity of currently available inanimate simulation models is questionable, because some tasks seem too far from clinical reality. The aim of this study was to validate a simulation model with six tasks commonly used in clinical practice (6-TSM) for the acquisition of psychomotor skills in minimally invasive surgery (MIS). METHODS: This was a prospective randomized trial comparing the 6-TSM to a previously described three-task training method (3 TTM). All first, second, and third postgraduate year surgical residents were eligible. The 6-TSM included clipping and dividing of a vessel, excision of lesion, appendectomy, mesh repair, suturing perforation, and hand-sewn anastomosis. The outcome measures of 6-TSM included accuracy error, tissue damage, sliding knot, leak, operating time, and dangerous movements. After completion of training, 6-TSM and 3-TTN residents were tested by the Minimally Invasive Surgical Trainer-Virtual Reality (MIST-VR). Criterion-related and construct validity, responsiveness, test-retest, and interrater reliability were assessed. RESULTS: During six months, 17 residents underwent training with the 6 TSM or the 3-TTM as allocated. The mean duration of training with 6-TSM and 3-TTM was similar (7.8 vs 8.1 h). The criterion-related validity of the 6-TSM was shown by significantly increased skill improvement in the 6-TSM residents, as compared with the 3-TTM residents at MIST-VR. Construct validity the of 6-TSM was shown by the finding that the experts' baseline was superior to the residents' baseline. The responsiveness of the 6-TSM was shown by the significantly increased skill improvement of the 6-TSM residents in sliding knot, leak, and operating time. The test-retest reliability of the 6-TSM was good (> 0.80), except for accuracy error and dangerous movements (Cronbach's intraclass correlation coefficient alpha: 0.57, p < 0.0001; 0.62, p < 0.0001, respectively). The interrater reliability of the 6-TSM was good (>0.80) except for leak (Kendall's concordance coefficient tau_b:0.76, p = 0.06 for hand-sewn anastomosis) and dangerous movements (tau_b:0.72, p = 0.08 for suturing perforation and tau_b:0.68, p = 0.10 for hand sewn anastomosis). The perresident cost for 6-TSM was 769 dollars. CONCLUSIONS: The 6-TSM is a valid and reliable learning tool for surgical residents' acquisition of laparoscopic motor skills. PMID- 15531972 TI - Assessing the benefits of "gaze-down" display location in complex tasks. AB - BACKGROUND: Location of the image display is one of several factors that influence perceptual processing and endoscopic manipulation in minimal access surgery. Previous studies have proved the benefits of the gaze-down stance, as compared with the conventional gaze-up stance. This study investigates the effect of the gaze-down stance on the performance of a task with varying manipulative and perceptual demands. METHODS: The participants in this study were 20 medical students. Each student performed endoscopic touching tasks under standard conditions using the Dundee Projection System (DPS) display, positioned to provide gaze-up and gaze-down stances. To increase task complexity, two kinds of manual coordination (unilateral vs bilateral) and three endoscope positions (different positions of misalignment) were used. The outcome measures were task execution time and number of errors. RESULTS: Overall, the gaze-down stance reduced time and errors, as compared with the gaze-up display. However, the benefit obtained from the gaze-down stance was more significant in the more difficult tasks (bilateral task and 90 degrees misalignments). CONCLUSIONS: The gaze-down stance reduces task time and errors, as compared with a gaze-up stance. The reduction in time and errors is more appreciable as task complexity increases. PMID- 15531973 TI - A stone extraction facilitation device to achieve an improved technique for performing LCBDE. AB - BACKGROUND: Laparoscopic common bile duct exploration (LCBDE) has proved to be a safe, cost-effective way to treat common bile duct (CBD) stones. Despite this, LCBDE has not gained widespread adoption by surgeons. The technique has proved difficult to master, and damage to the fragile choledochoscope by grasping forceps and passage through the port valves has been problematic. Cases involving large, impacted, or multiple stones have required conversion to open exploration of CBD. METHODS: The Multichannel Instrument Guide (MIG) is introduced as a solution for these problems. The MIG is a J-shaped plastic extrusion with three lumens. It is flexible and can be straightened for insertion through a 10-mm port. The MIG facilitates insertion of a flexible 2.8- to 3.2-mm choledochoscope into the CBD. At the same time, additional tools such as balloon or irrigation catheters and lithotripters can be introduced into the CBD. These can be manipulated under video guidance via the choledochoscope. This procedural multitasking allows for a more efficient LCBDE. The authors describe their initial experience using the MIG for 23 patients. RESULTS: Of the 23 patients who underwent LCBDE procedures, 20 had stones in the CBD. Multiple stones were present in 48% of the patients; impacted stones were present in 26% of the patients; and stones larger than 1 cm were present in 26% of the patients. A 95% stone clearance rate was achieved. Difficult cases with large, impacted or multiple stones were resolved using the MIG. Two choledochoscopes were damaged; one during surgery and one during cleaning and storage. CONCLUSIONS: The MIG has demonstrated significant advantages over previously described techniques. The device secures biliary tract access and allows procedural multitasking while protecting the delicate and expensive equipment. Subsequently, a simplified technique algorithm can be followed that may encourage more surgeons to adopt the routine performance of LCBDE. PMID- 15531974 TI - Treatment of severe chronic venous insufficiency using the subfascial endoscopic perforator vein procedure. AB - BACKGROUND: Before 1985, corrective surgical procedures for patients with chronic venous insufficiency and venous ulcers were performed using long incisions through diseased skin and subcutaneous tissues. The procedure, involving ligation of incompetent perforator veins, known as "the Linton operation," often was complicated by wound infections and poor healing. This changed in 1985 when G. Hauer demonstrated a new surgical technique for direct visual identification of the incompetent perforator veins using an endoscope in the subfascial space. This seminal contribution marked the advent of subfascial endoscopic perforator vein surgery (SEPS). METHODS: From 1996 to 2004, the authors group prospectively collected data on 110 patients with chronic venous insufficiency who underwent a SEPS procedure. Preoperative assessment of the limb's vascular status consisted of color-flow duplex ultrasound imaging and ascending and descending phlebography to locate vein valve incompetence, along with venous mapping. The ages of the patients ranged from 42 to 82 years (mean, 60 years). A total of 128 limbs underwent the SEPS procedure in the cohort of 110 patients. According to CEAP classification for venous limb disease, 60 limbs belonged to group C5 (skin changes, pigmentation, venous eczema, lipodermatosclerosis, healed ulcer) and 68limbs to group C6 (skin changes and active ulceration). RESULTS: The 110 patients underwent 128 SEPS procedures without significant morbidity. Of the 68 limbs in class C6, 54 showed ulcer healing within the follow-up period of 12 weeks. The remaining 14 limbs in class C6 achieved ulcer healing within 24 weeks. In this latter group, 10 patients had venous ulcers larger than 4 cm in diameter. These patients underwent a split-thickness skin graft at the time the SEPS procedure was performed. The grafts remained healed during a 2-year follow-up period. CONCLUSIONS: This study demonstrated the effectiveness of the SEPS procedure when incorporated into the overall treatment strategy for patients with chronic venous insufficiency. Minimal postoperative complications accompanied by ulcer healing and relief of lower extremity symptoms were achieved for all the patients, underscoring the important role of incompetent perforator veins in the formation of chronic venous insufficiency. PMID- 15531975 TI - Predicting baseline laparoscopic surgery skills. AB - BACKGROUND: Laparoscopic surgery requires specialized dexterity even beyond that required for open surgery. Decreased tactile feedback, different eye-hand coordination, and translation of a two-dimensional video image into a three dimensional working area are just some of the obstacles in the performance of laparoscopic surgery. Possession of certain nonsurgical skills may help in overcoming some of these obstacles. Prediction of baseline laparoscopic surgery skills may help further to refine the education of basic laparoscopic surgery skills. This investigation explores whether nonsurgical skills and demographic data can predict baseline laparoscopic surgery tasks. METHODS: First- and second year students were given a survey regarding nonsurgical dexterity skills. The survey inquired about typing skills, play with computer games, ability to sew, skill with music instruments, use of chopsticks, and experience operating tools. Demographic data were requested as well. All the students underwent four tasks: placing a piece of bowel in a retrieval bag, placing a stapler on the bowel, measuring a piece of bowel, and performing a liver biopsy in a porcine animal model. Both objective (time and error) and subjective evaluation were assessed for all the tasks. Statistical analysis using analysis of variances (ANOVA) Kruskal-Wallis test with post hoc tests, two-tailed unpaired t-tests/Mann-Whitney test, and Fischer's exact tests/chi-square tests was performed when appropriate. RESULTS: There were 68 students in this investigation. Gender, medical student year, ethnicity, desire to enter a surgical field, and age were not associated with increased performance in any of the tasks. Chopstick use was associated with statistically significantly better mean time in placing a piece of bowel in a retrieval bag and measuring a piece of bowel (p < 0.04). The other nonsurgical dexterity skills did not statistically increase performance, as indicated by time, errors, or subjective scores, for the four tasks. CONCLUSIONS: It is difficult to predict baseline laparoscopic surgery skills. PMID- 15531976 TI - The role of hand-assisted laparoscopic distal gastrectomy for distal gastric cancer. AB - BACKGROUND: The aim of this study was to evaluate the role of hand-assisted distal gastrectomy (HALDG) for gastric cancer. METHODS: This study prospectively enrolled 16 patients who underwent HALDG for early gastric cancer and matched them individually by sex, age, and body mass index to patients who underwent laparoscopically assisted distal gastrectomy (LADG) or open distal gastrectomy (ODG). Surgical outcomes were compared among the surgical methods. RESULTS: The mean operating time was the longest for the HALDG group, whereas wound size of the HALDG group was intermediate between that of the LADG and the ODG groups. The other surgical outcomes, such as the number of harvested lymph nodes, were not different among the groups. CONCLUSIONS: According to the findings, HALDG may not be as beneficial for patients with early gastric cancer as has been previously suggested. However, because of easier hand-eye coordination, HALDG may be an excellent bridge learning technique as a surgeon gains experience in laparoscopic gastrectomy. PMID- 15531977 TI - Does pregnancy begin at fertilization? PMID- 15531978 TI - Family physicians' beliefs about genetic testing. PMID- 15531979 TI - Breathing lessons. PMID- 15531980 TI - Public health educational intervention in a family medicine residency. AB - Emerging infections and bioterrorism pose increasing threats to the health of our communities. To protect patients, frontline providers like family physicians must learn and use core public health skills such as disease detection, surveillance, and response activities. This paper describes a unique approach to teaching these core public health skills by incorporating residents into ongoing disease surveillance and reporting activities in coordination with the local health department. Comparison of pre- and post-intervention surveys of residents demonstrated that this intervention increased resident self-perceived competency in disease detection and surveillance, knowledge of reportable conditions, and ability to engage response activities. PMID- 15531981 TI - Saved by an angel. PMID- 15531982 TI - The sagacity of persistence. PMID- 15531983 TI - Death pronouncements: surviving and thriving through stories. PMID- 15531984 TI - Can family medicine residents predict their performance on the in-training examination? AB - BACKGROUND AND OBJECTIVES: Evaluation of residents' competence is of great importance in the training of physicians, yet the evaluation process is largely subjective. Faculty and residents frequently rely on self assessments to make decisions regarding curriculum design and electives. The In-training Examination (ITE), the only widely available objective measure of residents' medical knowledge, provides an opportunity to test the reliability of self-assessments. This study's objective was to determine if family medicine residents are able to self-assess their medical knowledge by predicting their performance on the ITE. METHODS: A survey asking the residents to estimate their performance on the ITE in each of the nine content areas was administered at 13 examination sites just prior to the ITE. Correlation coefficients were calculated for corresponding predicted and actual scores for each resident in each content area. Predictions were also compared to performance according to quartile. RESULTS: Residents showed little ability to predict their scores in any of the content areas. Residents scoring in either the lowest or highest quartile were least able to predict accurately, with correct predictions ranging from 3% to 23%. CONCLUSIONS: Residents cannot reliably predict their performance on the ITE. Of special concern are residents scoring in the lowest quartile, since these residents greatly overestimated their performance. PMID- 15531985 TI - Which patients are most challenging for second-year medical students? AB - BACKGROUND AND OBJECTIVES: Fostering appropriate attitudes toward patients begins with recognition of the physician's personal biases and preferences about patients. We sought to assist second-year medical students to identify those patient attributes that may influence their clinical behavior, especially in difficult patient encounters. This study's objectives were (1) to identify the principal patient characteristics reported by students to evoke negative or positive affective reactions sufficient to compromise the quality of patient care and (2) to examine differences between male and female students on these patient characteristics. METHODS: An anonymous survey was made of all second-year medical students attending a seminar on medical errors. Students recorded the three principal patient characteristics that might evoke in them a negative personal reaction sufficient to compromise the quality of clinical care. Students then recorded three principal patient characteristics that might evoke a positive reaction sufficient to compromise the quality of clinical care. The data were analyzed to identify the primary concerns of the entire group of students and any significant differences between male and female students. RESULTS: Information on negative attitudes was provided by 67 of 71 (94%) students and on positive attitudes by 70 of 71 (99%). Content analysis identified four general categories of patient characteristics associated with negative attitudes (patients who are rude, contributed to medical problem, are abusive to others, and other attributes). Five general categories (patients with whom students have personal connection or patients who have physical and/or social attractiveness, vulnerability, power/influence, and other) were associated with positive attitudes by the students. The leading patient attributes reported as likely to influence clinical behavior were the positive attributes of "personal connection" and "attractiveness." These were reported by 60% and 53% of the class, respectively, compared to the leading negative attributes of "rudeness" and "contributed to own condition," each reported by 51% of students. Significant differences between male and female students only occurred in the categories of "rude" and "attractive." CONCLUSIONS: Students are willing to disclose specific, personal attitudes toward patients that they perceive as sufficiently serious to influence the quality of clinical care. More than half of the students anticipated difficulty in providing optimal care to patients who exhibit certain characteristics. More students reported concerns about positive affective reactions to patients than about negative reactions. Medical education should address these issues in strategies to adequately prepare students for practice. PMID- 15531986 TI - Medical student, physician, and public perceptions of health care disparities. AB - OBJECTIVES: This study investigates first- and fourth-year medical students' perceptions about health care disparities and compares their perceptions with those of physicians and the public. METHODS: We conducted an analysis of a national survey of medical students that included questions addressing unfair treatment of patients in the health care system based on insurance status, money, English language ability, and race/ethnicity. Results were compared with previously collected data from surveys of physicians and the public. The study also analyzed students' opinions about workforce diversity and cultural competence curricula. RESULTS: Medical students were generally more likely than physicians and the public to perceive unfair treatment of patients. First-year medical students were more likely than fourth-year students, and fourth-year students more likely than physicians, to perceive unfair treatment. Minority medical students and physicians were generally more likely than their white counterparts to perceive unfair treatment. The majority of medical students desired more exposure to disparity issues and endorsed medical workforce diversity. CONCLUSIONS: Perceptions of unfair treatment in the health care system differ among medical students, physicians, and the public, as well as among racial/ethnic groups. Minority students and physicians are more likely to perceive greater levels of unfairness. Our results suggest that perceptions of unfair treatment may decline during the process of acculturation to the medical profession. Interventions to reduce health care disparities must address the process of medical education and training. PMID- 15531987 TI - Physician informational needs in providing nutritional guidance to patients. AB - OBJECTIVES: This study's objective was to assess patient nutrition inquiries and related physician resources to identify the nutrition topics about which physicians most need information for patients and practice. METHODS: A survey was mailed to a 50% random sample of Washington Academy of Family Physicians (WAFP) members (n=778) to identify the top 10 patient nutrition inquiries that the physician would like to be better equipped to answer. The responding physicians' use of nutrition resources (journals, pamphlets, Web resources, toll-free numbers, and referrals to registered dietitians) was also recorded. RESULTS: A total of 306 (39.3%) physicians responded. The majority wanted more information on weight management (66.5%), followed by information on herbals, botanicals, and other complementary and alternative medicines (CAM) (36.4%) and vitamin and mineral supplements (24.4%). Other common nutrition inquiries for which respondents wanted more information included diet for prevention of disease, high protein-low carbohydrate diets, and childhood nutrition. A minority of respondents used nutrition Web resources and toll-free numbers. Nearly all respondents reported referring patients to registered dietitians. CONCLUSIONS: Physicians in our survey expressed a need for information on topics such as obesity, CAM, and life cycle and disease-specific diets. Medical nutrition educators should consider the educational needs of the practicing physician. PMID- 15531988 TI - Moving from undiagnosed to diagnosed diabetes: the patient's perspective. AB - BACKGROUND AND OBJECTIVES: One third of diabetes cases in the United States are undiagnosed. Knowledge of the patient's experience from symptom recognition to diabetes diagnosis will help clinicians and policy makers optimize their approach to diabetes detection. METHODS: We interviewed 15 patients diagnosed with type 2 diabetes within the past 6 months about how they came to be diagnosed and about any barriers that might have prevented their diagnosis. We used grounded theory qualitative methods to approach and analyze the semi-structured interviews. RESULTS: Most diagnoses of diabetes in these patients were either serendipitous, symptom driven, or patient initiated. None resulted from physician-initiated screening. Patients had only a superficial knowledge of the symptoms of diabetes prior to diagnosis, despite strong family histories of diabetes. Patients often incorrectly attributed symptoms of diabetes to other causes and sometimes physicians also did this. Barriers of cost, insurance, and trust were not deemed to be important by these patients. CONCLUSIONS: Many individuals with undiagnosed diabetes are likely unaware of the relevance of their symptoms. Clinicians must be vigilant in identifying people at risk for diabetes. Improved education of individuals at risk for diabetes may be a useful strategy to increase diabetes detection. PMID- 15531989 TI - Exploring the organizational culture of exemplary community health center practices. AB - BACKGROUND AND OBJECTIVES: A 1999 precursor to this study, published in Family Medicine, suggested that the organizational culture of practices was potentially important to health care providers and patients. In this research, we examined the experiences of people working in exemplary community health center practices to explore the components and maintaining factors of positive medical organizational culture. METHODS: Two exemplary practices were identified through a process of nominations and selection with respect to a presumptive definition of positive organizational culture. Interpretive categories and themes were developed through qualitative content analysis of semi-structured interviews, along with field observation. RESULTS: Categories of culture in these practices included Community Mission and Values, Leadership and Organizational Dynamics, Relationships, and Physical Space. Cultural qualities were nurtured by leadership approaches, collaborative staff meetings, and shared values about mission and workplace relationships. Staff consistently indicated that the spirit or culture in their practices was beneficial for employees, patients, and the process of clinical care. CONCLUSIONS: The positive organizational culture in these practices was substantially characterized and cultivated by specific values, attitudes, behaviors, and relationships of employees. Further work is indicated in approaches to assessment and intervention with organizational culture in medical settings and in evaluating associations with medical outcomes. PMID- 15531990 TI - An international physician education program to support the recent introduction of family medicine in Egypt. AB - BACKGROUND AND OBJECTIVES: There are few reports of systematic international physician development programs to create family medicine as a new specialty in a developing nation. This paper describes the process and outcomes of a large-scale effort to initiate new family medicine training through the Egyptian Ministry of Health and Population (MOHP) using a 12-week US-based program at the University of California, Irvine (UCI). METHODS: Generalist physicians (n=134) with 1 year of internship training, currently working under the MOHP in Egypt, were competitively selected to participate in a training program at UCI between 1998 and 2002. Participants were assessed before, during, and after the program using multiple measures of competencies in family medicine topics, practice, and teaching. Aggregate participant data, post-program quality surveys, and follow-up surveys of the program's influence on practice behaviors comprised the main measures used for program evaluation. RESULTS: Participants showed improvement in knowledge and skills for family medicine practice and teaching for topics covered in the program. After returning to Egypt, 98% reported continued use of their newly acquired skills and knowledge. Participants reported that the program advanced their careers, they taught family medicine to other physicians, and they were likely to pursue certification under a newly established Family Medicine Board of Egypt. Self-reported practice in family medicine increased to 69% after the program versus 16% before. CONCLUSIONS: Overseas training programs are a viable method of introducing family medicine as a new clinical specialty. Ingredients for successful implementation and barriers are discussed. PMID- 15531997 TI - Hemoglobin mass and peak oxygen uptake in untrained and trained female altitude residents. AB - Total hemoglobin mass has not been systematically investigated in females at altitude. We measured this quantity (CO-rebreathing method) as well as peak oxygen uptake in 54 young women (age 22.5 +/- 0.6 SE years) with differing physical fitness living in Bogota (2600 m) and compared the results with those of 19 subjects from 964 m in Colombia and 75 subjects from 35 m in Germany. In spite of an increased hemoglobin concentration the hemoglobin mass was not changed in highlanders (means 9.0 to 9.5 g . kg (-1) in untrained subjects at all altitude levels). Endurance trained athletes, however, showed a rise in hemoglobin mass by 2 - 3 g . kg (-1) at all sites. Erythropoietin was little increased in Bogota; iron stores were within the normal range. Aerobic performance capacity was lower at high altitude than at sea level and remained so also after correction for the hypoxic deterioration in untrained and moderately trained subjects but not in athletes; possibly the cause was reduced daily physical activity in non-athletic Bogotanians compared to lowlanders. After exclusion of the factor V.O(2peak) by analysis of covariance a mean rise of 6.6 % in hemoglobin mass at 2600 m was calculated being smaller than in males (> 12 %). The attenuated increase of hemoglobin mass in female highlanders possibly results from stimulation of ventilation improving arterial oxygen saturation or from an increased hypoxia tolerance of cellular metabolism both caused by female sexual hormones. PMID- 15531998 TI - Is hemoglobin desaturation related to blood viscosity in athletes during exercise? AB - Several studies have suggested that athletes with low hemoglobin saturation during exercise may experience impaired pulmonary blood gas exchange during maximal exercise. Blood viscosity may be implicated in exercise-induced pulmonary hemorrhage in race horses. We hypothesized that blood rheology may contribute to impaired gas exchange and reduced hemoglobin saturation during exercise in humans. A group of 20 highly trained endurance athletes participated in this study, 9 with low hemoglobin saturation during exercise (Low-SpO (2) group) and 11 with normal hemoglobin saturation (High-SpO (2) group). All subjects performed a progressive exercise test conducted to V.O (2max). Venous blood was sampled at rest, 50 % V.O (2max) and maximal exercise. Blood viscosity (etab) was measured at very high shear rate (1000 s (-1)) and 37 degrees C with a falling ball viscometer. The erythrocyte rigidity coefficient, "Tk", was calculated using the Dintenfass equation. At rest, no significant difference in etab was observed between the two groups (3.00 +/- 0.08 mPa . s vs. 3.01 +/- 0.04 mPa . s for the Low-SpO (2) and High-SpO (2) group, respectively). At 50 % V.O (2max) and maximal exercise, etab was higher in Low-SpO (2) (p < 0.01). Tk decreased in High-SpO (2) (p < 0.01) but remained unchanged in the other group during testing. The greater increase in etab in the Low-SpO (2) group during exercise may therefore have been due to the lack of reduction in Tk. As suggested by previous studies, the greater increase in blood viscosity in athletes with low hemoglobin saturation may lead to vascular shear stress. Whether this could impair the blood gas barrier and result in exercise-induced hypoxemia requires further study. PMID- 15531999 TI - Brain magnetic resonance imaging, aerobic power, and metabolic parameters among 30 asymptomatic scuba divers. AB - The aim of the study was to evaluate the presence of cerebral lesions in asymptomatic scuba divers and explain the causes of them: potential risk factors associating cardiovascular risk factors, low aerobic capacity, or characteristics of diving (maximum depth, ascent rate). Experienced scuba divers, over 40 years of age, without any decompression sickness (DCS) history were included. We studied 30 scuba divers (instructors) without any clinical symptoms. For all of them, we carried out a clinical examination with fatty body mass determination and we questioned them about their diving habits. A brain Magnetic Resonance imaging (MRI), an assessment of maximal oxygen uptake, glycemia, triglyceridemia, and cholesterolemia were systematically carried out. Cerebral spots of high intensity were found at 33 % in the scuba diving group and 30 % in the control group. In the diving group, abnormalities were related to unsafe scuba-diving or metabolic abnormalities. In our study, we did not find a significant relationship between the lesions of the central nervous system, and the age, depth of the dives, number of dives, and ergometric performances (maximal oxygen uptake, V.O (2max), serum level of blood lactate). Nevertheless, we found a significant relationship between the lesions of the central nervous system and ascent rate faster than 10 meters per minute (r = 0.57; p = 0.003) or presence of high level of cholesterolemia (r = 0.6; p = 0.001). We found concordant results using the Cochran's Test: meaningful link between the number of brain lesions and the speed of decompression (Uexp = 14 < Utable = 43; alpha = 0.05, p < 0.01). We concluded that hyperintensities can be explained by preformed nitrogen gas microbubbles and particularly in presence of cholesterol, when the ascent rate is up to 10 meters per minute. So, it was remarkable to note that asymptomatic patients practicing scuba diving either professionally or recreationally, presented lesions of the central nervous system. This survey permitted us to highlight in a population of professional divers, neurological and also cardiovascular abnormalities (ventricular arrhythmias); although none of them present any symptoms today. It seems therefore important to us to propose in the future, for a better prevention of neurological injuries, a systematic follow-up by maximal oxygen consumption measure, brain MRI, and cholesterolemia. In the same way, our results suggest a modification of the diving tables with a maximal decompression rate at 9 m . mn ( 1). PMID- 15532001 TI - Treatment of muscle injuries by local administration of autologous conditioned serum: a pilot study on sportsmen with muscle strains. AB - Muscle injuries represent a major part of sports injuries and are a challenging problem in traumatology. Strain injuries are the most common muscle injuries after contusions. These injuries can lead to significant pain and disability causing time to be lost to training and competition. Despite the frequency of strain injuries the treatment available is limited and is generally not sufficient to enhance muscle regeneration efficiently when fast resumption of sport activity is a primary target. A number of growth factors play a specific role in regeneration and it has been proven that a previously described method of physically and chemically stimulating whole blood (to produce autologous conditioned serum) induces concentration increases in FGF-2, HGF, and TGF-beta1. A preliminary study was conducted on muscle strain injuries in professional sportsmen receiving either: 1. autologous conditioned serum (ACS) or 2. Actovegin/Traumeel treatment as control. Assessment of recovery from injury was done by: 1. sport professional's ability to participate to 100 % under competition conditions in their respective sport and 2. MRI analysis. A significant difference in the recovery time from injury was demonstrated: 16.6 +/ 0.9 in the ACS treated instead of 22.3 +/- 1.2 (mean +/- SEM) days in the Actovegin/Traumeel control group (p = 0.001). MRI analysis supported the observed acceleration of the lesion recovery time. We conclude that ACS injection is a promising approach to reduce the time to recovery from muscle injury. PMID- 15532000 TI - Treatment of muscle injuries by local administration of autologous conditioned serum: animal experiments using a muscle contusion model. AB - Muscle contusions represent a major part of sports injuries. The suggested treatments are generally sufficient to support muscle healing, but require a relatively long period of time. Given that autologous blood products are safe treatments, we have used a technique which stimulates the release of certain growth factors in the autologous conditioned serum (ACS). Those growth factors are known to improve the proliferative activity of myogenic precursor cells. Mice were subjected to an experimental contusion injury to their gastrocnemius muscle; one group received local injections of ACS at 2 hrs, 24 hrs, and 48 hrs after injury, a control group received saline injections. The histology results showed that satellite cell activation at 30/48 hrs post injury was accelerated and the diameter of the regenerating myofibers was increased compared to the controls within the first week after injury. ELISA results on the ACS have shown that the elevations in FGF-2 (460 %) and TGF-beta1 (82 %) could be partly responsible for the accelerating effects on regeneration due to proliferative and chemotactic properties. We conclude that ACS injection is a promising approach to reduce the time of recovery from muscle injury. In terms of clinical targets, this new approach could be used in the treatment of sports injuries and may also be interesting in postoperative situations. PMID- 15532002 TI - A cross-sectional analysis of sagittal knee laxity and isokinetic muscle strength in soccer players. AB - In this cross-sectional study, sagittal knee laxity and isokinetic strength of knee extensor and flexor muscle groups were measured and differences related to leg dominance were evaluated. A total of 44 healthy male soccer players (who had trained regularly at least for the last five years) and 44 sedentary people as their control counterparts were involved in this study. All participants were tested using a KT-1000 knee arthrometer for knee laxity. Isokinetic concentric knee peak torque and hamstring/quadriceps (H/Q) ratio were also measured at 60, 180, 300 degrees/s through a Cybex 2 - 340 dynamometer. Posterior laxity in the non-dominant side of soccer players was significantly higher than in the dominant side (p < 0.005) while there were no significant anterior and total anteroposterior (total AP) laxity differences in both groups. Soccer players had significantly lower anterior and total AP laxity values than controls (p < 0.0001) while there was no significant difference between posterior laxity values in both sides. Dominant extremity demonstrated significantly higher knee flexor peak torque and H/Q ratio at 180 degrees /s in soccer players (p < 0.05). Similarly in sedentary controls, H/Q ratio at 60 degrees /s of the dominant side was significantly higher than that in the non-dominant side (p < 0.05). Soccer players had significantly higher extensor and flexor peak torque values and H/Q ratios than sedentary subjects for both extremities. In both groups, there were no significant correlations between knee laxity and isokinetic knee extensor and flexor strength and H/Q ratios except weak negative correlation between posterior knee laxity and isokinetic extensor peak torque at 60, 180 and 300 degrees /s (p < 0.005, r = - 0.43, p < 0.05, r = - 0.39, p < 0.05, r = - 0.32 respectively) in the non-dominant side of soccer players and at 300 degrees /s (p < 0.05, r = - 0.32) in the non-dominant side of controls. Soccer players demonstrated significantly less sagittal knee laxity and higher isokinetic strength of the knee flexors and extensors compared to sedentary controls. Isokinetic strength difference was found to be higher for the flexor muscle group. Further prospective studies are needed to explain whether the increased H/Q ratio decreases the risk of ligamentous injury. PMID- 15532003 TI - Central adiposity, aerobic fitness, and blood pressure in premenopausal Hispanic women. AB - Hispanics comprise one of the fastest-growing segments of the U.S. population. Mexican-American adults are more likely to be overweight, physically inactive, diabetic, and to have higher levels of hypertension than are white adults. However, studies addressing the relationship between physical fitness and coronary artery disease (CAD) risk factors among Mexican-Americans are much less conclusive. Therefore, understanding the etiology of factors influencing resting systolic (SBP) and diastolic blood pressure (DBP) in Hispanic women was the aim of this investigation. SBP, DBP, peak oxygen uptake (peak VO (2)), weekly physical activity, waist (WC) and hip circumference, blood glucose, and levels of plasma lipids (triglyceride, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein-cholesterol) of 39 Hispanic women age 22 - 51 years were measured. Factors with significant correlation to SBP were age, WC, sagittal diameter, and weight. Similarly, significant correlations were observed between anthropometric indices, age, and DBP. Peak VO (2) ( r = - 0.53, p < 0.01) and heart rate at maximal effort ( r = - 0.34, p 0.05) between test when the amount of external work completed was taken into consideration. There was no systematic bias (p > 0.05) for mean (+/- s) peak values of V.O (2) (3.12 [0.37] vs. 3.04 [0.38] L . min (-1)) or any other parameter between the step and ramp tests, respectively. Mean values of T (lim), PMP, and TWD were also similar (p > 0.05) between tests. However, the level of agreement for peak values of all test parameters was low. It is therefore concluded that while either test can be considered as being suitable for the purpose of eliciting V.O (2) peak and other physiological responses using ACE, they should not be used interchangeably for the purpose of assessing parameters linked to the aerobic capacity of the upper-body. PMID- 15532007 TI - A physiological comparison of synchronous and asynchronous hand cycling. AB - The purpose of this study was to compare submaximal physiological responses (oxygen uptake, ventilation, heart rate) and gross mechanical efficiency between synchronous and asynchronous hand cycling at different cadences. Thirteen non disabled men (22.4 +/- 1.6 yr) performed two submaximal exercise tests on a treadmill, using synchronous and asynchronous crank settings in counter balanced order. Tests were performed using a commercially available hand cycle unit that was attached to a hand rim wheelchair. Each test consisted of five 5-min exercise bouts at 36, 47, 55, 65, and 84 rpm. ANOVA for repeated measures showed a significant effect of crank mode (p < 0.001) and cadence (p < 0.001), as well as an interaction effect between both (p < 0.01). Physiological responses were lower, and efficiency higher, in synchronous versus asynchronous hand cycling at all cadences. Post-hoc analysis of the (overall) effect of cadence showed significantly higher physiological responses and lower efficiency at the higher (84 vs. 65 rpm and 65 vs. 55 rpm) and lower (36 vs. 47 rpm) cadences. The interaction effect indicates that the effect of crank mode was dependent on cadence, showing a larger difference between synchronous and asynchronous hand cycling at 84 vs. 65 rpm and at 36 vs. 47 rpm. It is concluded that, in contrast to previous results in arm crank ergometry, synchronous hand cycling is less strenuous and more efficient than asynchronous hand cycling. PMID- 15532008 TI - Relationship between diet and serum anabolic hormone responses to heavy resistance exercise in men. AB - Relationship between dietary intake and serum anabolic hormone concentrations of testosterone (T), free testosterone (FT), and growth hormone were examined at rest as well as after the heavy-resistance exercise (HRE) in 8 strength athletes (SA) and 10 physically active non-athletes (NA). In the first part of the study serum basal anabolic hormone concentrations and dietary intake were examined in the total group of subjects. In the second part of the study a subgroup of 5 SA and 5 NA performed the high volume and high intensity HRE. Dietary intake was registered by dietary diaries for 4 days preceding the loading day. Significant correlations were observed between serum basal T and fat (E%: r = 0.55, p < 0.05, g/kg: r = 0.65, p < 0.01) and protein intake (E%: r = - 0.77, p < 0.001, g/kg: r = - 0.68, p < 0.01) in the total group of subjects. However, when the two groups were examined separately the significant relationships between serum basal T and dietary fat and protein could be noticed in SA only (fat g/kg: SA r = 0.77, p < 0.05; in NA r = 0.44, n.s., protein g/kg: SA r = - 0.84, p < 0.05; in NA r = 0.27, n.s.). Both serum T and FT responses to HRE were correlated with fat (E%: r = 0.85, p < 0.01 and r = 0.73, p < 0.05, g/kg: r = 0.72, p < 0.05 and r = 0.77, p < 0.01) and protein (E%: r = - 0.81, p < 0.01 and r = - 0.69, p < 0.05, g/kg: r = - 0.86, p < 0.01 and r = - 0.65, p < 0.05). The results suggest the possible role of diet leading to alterations in serum T and FT during prolonged strength training, and that diets with insufficient fat and/or excessive protein may compromise the anabolic hormonal environment over a training program. PMID- 15532009 TI - Maximal resultant four fingertip force and fatigue of the extrinsic muscles of the hand in different sport climbing finger grips. AB - This study investigates the effect of simulated sport climbing finger grips on the resultant four fingertip force and the rate of fatigue of finger flexor muscles. Six elite sport climbers sat on a chair with the right forearm placed in a handgrip dynamometer modified so that only the fingertips applied direct force. They were asked to perform three maximal voluntary contractions (MVC). After ten minutes, they had to reach 80 % of the peak MVC intermittently with a 5 s contraction followed by 5 s of rest for 20 repetitions. Two common sport climbing finger grips were tested: the "slope" grip and the "crimp" grip. In the "crimp" grip, the distal interphalangeal joint (DIP) is hyper-extended and the proximal interphalangeal joint (PIP) is flexed from 90 degrees to 100 degrees . In the "slope" grip, DIP is flexed from 50 degrees to 70 degrees and PIP is flexed just slightly. The surface EMG of the hand extrinsic flexors and the maximal resultant four fingertip force were recorded. Results show that the maximal resultant four fingertip force does not depend on the type of finger grips (on average 420 N, p > 0.05). EMG median frequency of finger flexor muscles and resultant four fingertip force rate decrease are similar between both sport climbing finger grips (p > 0.05). This shows that the fatigue rate is not dependent upon the sport climbing finger grips. In conclusion, the results suggest that the use of the "crimp" or the "slope" grip does not provide any benefit with respect to muscular fatigue in sport climbing. PMID- 15532010 TI - Dermatological marks in athletes of artistic and rhythmic gymnastics. AB - The authors present dermatological signs in: a) rhythmic gymnastics athletes, b) male artistic gymnastics athletes, compared to a control group of fitness athletes. Athletes from the artistic gymnastics group were observed twice. The signs they showed on their first examination (20 days previous to the competition) were two circular zones of thickening of the skin with relation to the radial epiphysis. In all of them, two zones of frictional alopecia were present, one on the dorsal face of the forearms, slantwise outlined, the other on the wrists. A noticeable thickening of the skin was present on the palms of the hands. On a second examination, at the beginning of the training, after about two months of inactivity, the alopecic area was replaced by hypertrichosis, although featuring different patterns in each athlete. Thickening of the skin was slightly smaller than that observed at the first examination. The authors describe onychopathology shown in its different forms in 94 % of the athletes of the rhythmic group. Subsequently the authors discuss the pathogenesis of the above described signs. PMID- 15532011 TI - A crucial point: the early diagnosis of increased risk subjects before sudden cardiac death. PMID- 15532018 TI - Controllable donor-acceptor neutral [2]rotaxanes. AB - In pursuit of a neutral bistable [2]rotaxane made up of two tetraarylmethane stoppers--both carrying one isopropyl and two tert-butyl groups located at the para positions on each of three of the four aryl rings--known to permit the slippage of the pi-electron-donating 1,5-dinaphtho[38]crown-10 (1/5DNP38C10) at the thermodynamic instigation of pi-electron-accepting recognition sites, in this case, pyromellitic diimide (PmI) and 1,4,5,8-naphthalenetetracarboxylate diimide (NpI) units separated from each other along the rod section of the rotaxane's dumbbell component, and from the para positions of the fourth aryl group of the two stoppers by pentamethylene chains, a modular approach was employed in the synthesis of the dumbbell-shaped compound NpPmD, as well as of its two degenerate counterparts, one (PmPmD) which contains two PmI units and the other (NpNpD) which contains two NpI units. The bistable [2]rotaxane NpPmR, as well as its two degenerate analogues PmPmR and NpNpR, were obtained from the corresponding dumbbell-shaped compounds NpPmD, PmPmD, and NpNpD and 1/5DNP38C10 by slippage. Dynamic 1H NMR spectroscopy in CD2Cl2 revealed that shuttling of the 1/5DNP38C10 ring occurs in NpNpR and PmPmR, with activation barriers of 277 K of 14.0 and 10.9 kcal mol(-1), respectively, reflecting a much more pronounced donor-acceptor stabilizing interaction involving the NpI units over the PmI ones. The photophysical and electrochemical properties of the three neutral [2]rotaxanes and their dumbbell-shaped precursors have also been investigated in CH2Cl2. Interactions between 1/5DNP38C10 and PmI and NpI units located within the rod section of the dumbbell components of the [2]rotaxane give rise to the appearance of charge-transfer bands, the energies of which correlate with the electron accepting properties of the two diimide moieties. Comparison between the positions of the visible absorption bands in the three [2]rotaxanes shows that, in NpPmR, the major translational isomer is the one in which 1/5DNP38C10 encircles the NpI unit. Correlations of the reduction potentials for all the compounds studied confirm that, in this non-degenerate [2]rotaxane, one of the translational isomers predominates. Furthermore, after deactivation of the NpI unit by one-electron reduction, the 1/5DNP38C10 macrocycle moves to the PmI unit. Li+ ions have been found to strengthen the interaction between the electron donating crown ether and the electron-accepting diimide units, particularly the PmI one. Titration experiments show that two Li+ ions are involved in the strengthening of the donor-acceptor interaction. Addition of Li+ ions to NpPmR induces the 1/5DNP38C10 macrocycle to move from the NpI to the PmI unit. The Li+ ion-promoted switching of NpPmR in a 4:1 mixture of CD2Cl2 and CD3COCD3 has also been shown by 1H NMR spectroscopy to involve the mechanical movement of the 1/5DNP38C10 macrocycle from the NpI to the PmI unit, a process that can be reversed by adding an excess of [12]crown-4 to sequester the Li+ ions. PMID- 15532013 TI - Malic enzyme 2 may underlie susceptibility to adolescent-onset idiopathic generalized epilepsy. AB - Idiopathic generalized epilepsy (IGE) is a class of genetically determined, phenotypically related epilepsy syndromes. Linkage analysis identified a chromosome 18 locus predisposing to a number of adolescent-onset IGEs. We report a single-nucleotide polymorphism (SNP) association analysis of the region around the marker locus with the high LOD score. This analysis, which used both case control and family-based association methods, yielded strong evidence that malic enzyme 2 (ME2) is the gene predisposing to IGE. We also observed association among subgroups of IGE syndromes. An ME2-centered nine-SNP haplotype, when present homozygously, increases the risk for IGE (odds ratio 6.1; 95% confidence interval 2.9-12.7) compared with any other genotype. Both the linkage analysis and the association analysis support recessive inheritance for the locus, which is compatible with the fact that ME2 is an enzyme. ME2 is a genome-coded mitochondrial enzyme that converts malate to pyruvate and is involved in neuronal synthesis of the neurotransmitter gamma-aminobutyric acid (GABA). The results suggest that GABA synthesis disruption predisposes to common IGE and that clinical seizures are triggered when mutations at other genes, or perhaps other insults, are present. PMID- 15532019 TI - Complete charge pooling is prevented in viologen-based dendrimers by self protection. AB - We have investigated the electrochemical behavior, and chemical and photosensitized reduction of two dendrimers based on a 1,3,5-trisubstituted benzenoid core, which contain 9 and 21 4,4'-bipyridinium (usually called viologen) units, respectively, in their branches and are terminated with tetraarylmethane groups. For comparison purposes, the behavior of reference compounds that contain a single viologen unit have also been investigated. We have found that only part of the viologen units can be reduced in the dendrimer species. For the larger dendrimer, the number of reducible viologens (out of the 21 present) is 14 in electrochemical experiments (in MeCN), 9 on reduction with bis(benzene)chromium (in MeCN), and 13 by photoinduced electron transfer with 9 methylanthracene as a photosensitizer and triethanolamine as a sacrificial reductant in CH2Cl2. The reduced viologen units undergo partial dimerization. The photochemical experiments have shown that only monomeric, one-electron-reduced viologen units are formed at the beginning of the irradiation, followed by dimer formation, until a photostationary state is reached that contains 40 % nonreduced, 33 % monomeric reduced, and 27 % reduced units associated in the dimeric form. The results suggest that, upon reduction of a fraction of the viologen units, the dendrimer structure shrinks, with the result that the bulky terminal groups protect other viologen units from being reduced. PMID- 15532020 TI - Synthesis of branched oxime-linked peptide mimetics of the MUC1 containing a universal T-helper epitope. AB - Our goal was to develop mimics of MUC1, highly immunogenic to induce an efficient immune response against the tumor-associated form of MUC1, and sufficiently different from the natural antigen to bypass the tolerance barrier in humans. With the aim of obtaining a well-defined peptide construct as a means of evoking the precise immune responses required in immunotherapy, we synthesized artificial mimics of the MUC1 protein composed of two MUC1 repeat units of inverse orientation and a universal T-helper epitope. To synthesize these heteromeric peptide constructs, we followed a convergent approach using chemoselective ligation based on oxime chemistry. A stem peptide was first synthesized bearing two orthogonally masked aldehydes. After successive deprotection, two oxime bonds can be specifically generated. The proposed strategy proved to be concise and robust, and allowed the synthesis of the tri-branched protein in a very satisfactory yield. The different constructs were tested for their ability to generate antibodies able to recognize the MUC1 protein. PMID- 15532021 TI - Morphology of mitochondrial permeability transition: morphometric volumetry in apoptotic cells. AB - Here we report on the mitochondrial permeability transition (MPT), which refers to the morphology of mitochondria whose inner membrane has lost its selective permeability. In all types of apoptotic cells so far examined, we found outer mitochondrial membranes that had been ruptured. These mitochondria present a swollen matrix covered by an inner membrane herniating into the cytoplasm through the breached outer membrane. Similarly ruptured outer mitochondrial membranes have been reported in studies on mitochondrial fractions induced to undergo MPT, carried out by others. Our observations were made on five types of rat tissue cells and six different cultured cell lines in the early stages of apoptosis. Samples from the cell lines HL-60, HeLa, WEHI-164, and a special batch of PC-12 cells were subjected to various apoptogenic agents and analyzed morphometrically. Nonapoptotic companion cells with unaltered nuclear structure (CUNS) were also analyzed. The mitochondrial volume in microm(3) and the volume fraction of the cytoplasm occupied by mitochondria in cells with typical nuclear signs of apoptosis and also in CUNS were evaluated. The volume of the mitochondria with ruptured membrane represents at least 69% (47-89%) of the total mitochondrial volume of the apoptotic cells. Thus, a considerable fraction of the cellular mitochondrial mass is or was in the state of permeability transition and probably involved in enhancement of the apoptotic program. In all samples, a fraction of the cells with normal nuclei possessed mitochondria with breached outer membranes as described above. In these cells, MPT occurred before the appearance of the typical nuclear phenotype of the apoptotic cells. PMID- 15532022 TI - Six novel alleles identified in Italian hereditary fructose intolerance patients enlarge the mutation spectrum of the aldolase B gene. AB - Hereditary fructose intolerance (HFI) is a recessively inherited disorder of carbohydrate metabolism caused by impaired functioning of human liver aldolase (B isoform; ALDOB). To-date, 29 enzyme-impairing mutations have been identified in the aldolase B gene. Here we report six novel HFI single nucleotide changes identified by sequence analysis in the aldolase B gene. Three of these are missense mutations (g.6846T>C, g.10236G>T, g.10258T>C), one is a nonsense mutation (g.8187C>T) and two affect splicing sites (g.8180G>C and g.10196A>G). We have expressed in bacterial cells the recombinant proteins corresponding to the g.6846T>C (p.I74T), g.10236G>T (p.V222F), and g.10258T>C (p.L229P) natural mutants to study their effect on aldolase B function and structure. All the new variants were insoluble; molecular graphics data suggest this is due to impaired folding. PMID- 15532023 TI - Large BRCA1 gene deletions are found in 3% of German high-risk breast cancer families. AB - We have tested for large BRCA1 gene rearrangements in German high-risk breast and ovarian cancer families previously screened negative for point mutations by dHPLC and sequencing. Using the novel MLPA method, two deletions of exons 1A, 1B and 2 and exon 17, respectively, were detected in four out of 75 families investigated in Southern Germany. An identical exon 17 deletion with the same breakpoints and a deletion of exons 1A, 1B and 2 were found by fluorescent multiplex PCR in two out of 30 families investigated in Northern Germany. Combining both populations, genomic rearrangements were found in 6% of the mutation-negative families and 3% of all high-risk families and account for 8% of all BRCA1 mutations. Our data indicate that the exon 17 deletion may be a founder mutation in the German population. The prevalence of BRCA1 gene deletions or duplications in our patients is similar to previous reports from Germany and France. Genomic quantification by MLPA is a useful method for molecular diagnostics in high-risk breast cancer families. PMID- 15532024 TI - Mutations of the Nogo-66 receptor (RTN4R) gene in schizophrenia. AB - Schizophrenia (SCZD) or schizoaffective disorders are quite common features in patients with DiGeorge/velo-cardio-facial syndrome (DGS/VCFS) as a result of chromosome 22q11.2 aploinsufficiency. We evaluated the Nogo-66 receptor gene (RTN4R), which maps within the DGS/VCFS critical region, as a potential candidate for schizophrenia susceptibility. RTN4R encodes for a functional cell surface receptor, a glycosylphosphatidylinositol (GPI)-linked protein, with multiple leucine-rich repeats (LRR), which is implicated in axonal growth inhibition. One hundred and twenty unrelated Italian schizophrenic patients were screened for mutations in the RTN4R gene using denaturing high performance liquid chromatography (DHPLC). Three mutant alleles were detected, including two missense changes (c.355C>T; R119W and c.587G>A; R196H), and one synonymous codon variant (c.54G>A; L18L). The two schizophrenic patients with the missense changes were strongly resistant to the neuroleptic treatment at any dosage. Both missense changes were absent in 300 control subjects. Molecular modeling revealed that both changes lead to putative structural alterations of the native protein. PMID- 15532025 TI - Identification and detection of the periostin gene in cardiac development. AB - Periostin, a member of the fasciclin gene family, acts as a cell adhesion molecule through binding to cell surface integrins. Periostin expression has previously been shown to increase substantially following transforming growth factor beta (TGF-beta) and bone morphogenetic protein stimulation. As these molecules are indispensable for cardiac development, we sought to clone the chicken ortholog of periostin and evaluate its spatiotemporal expression pattern during heart morphogenesis. We show by Northern analysis, whole mount and section in situ hybridization experiments that periostin is predominantly expressed in the developing endothelium of the ventricular trabeculae as well as in the endothelium and mesenchyme of the outflow tract and atrioventricular endocardial cushions. Cardiac expression continues into fetal development where periostin is seen predominantly in the valve leaflets and supporting chordae tendinae. PMID- 15532026 TI - A novel mutation in UDP-N-acetylglucosamine-1-phosphotransferase gamma subunit (GNPTAG) in two siblings with mucolipidosis type III alters a used glycosylation site. AB - The N-acetylglucosaminyl-1-phosphotransferase (termed phosphotransferase) catalyzes the initial step in the formation of mannose 6-phosphate (M6P) residues required for the efficient transport of soluble lysosomal enzymes. The phosphotransferase is a multisubunit enzyme composed of three subunits (alpha2beta2gamma2) that are products of two genes. The gene encoding the gamma subunit (GNPTAG) appears to be defective in patients with mucolipidosis type III (ML III). We have analyzed the GNPTAG gene in two siblings with ML III showing elevated activities of several lysosomal enzymes in cultured fibroblasts serum and diminished activities in cultured fibroblasts. Immunoprecipitation of metabolically labeled cathepsin D (CtsD) from fibroblasts revealed that the sorting/transport of this lysosomal protease was affected. Addition of ammonium chloride inhibiting pH-dependent processes, such as the CtsD-M6P receptor interaction, indicated that 15 to 20% of the newly synthesized CtsD is transported in ML III fibroblasts in an M6P-dependent manner. By direct sequencing a novel homozygous mutation, c.347_349delACA (p.Asn116del), was identified affecting a potential N-linked glycosylation site. Western blot analysis of extracts from control fibroblasts detect a 97 kDa glycosylated dimer whereas ML III cells contain a GNPTAG dimer of reduced molecular mass. These data suggest that the loss of the used glycosylation site in the gamma subunit may affect the intracellular localization of GNPTAG and the overall efficiency of M6P formation. PMID- 15532027 TI - Chemistry challenges in SNP typing. PMID- 15532028 TI - Modified microperoxidases exhibit different reactivity towards phenolic substrates. AB - The reactivity of several microperoxidase derivatives with different distal-site environments has been studied. The distal-site environments of these heme peptides include a positively charged one, an uncharged environment, two bulky and doubly or triply positively charged ones, and one containing aromatic apolar residues. The reactivity in the catalytic oxidation of two representative phenols, carrying opposite charges, by hydrogen peroxide has been investigated. This allows the determination of the binding constants and of the electron transfer rate from the phenol to the catalyst in the substrate/microperoxidase complex. The electron-transfer rates scarcely depend on the redox and charge properties of the phenol, but depend strongly on the microperoxidase. Information on the disposition of the substrate in the adducts with the microperoxidases has been obtained through determination of the paramagnetic contribution to the 1H NMR relaxation rates of the protons of the bound substrates. The data show that the electron-transfer rate drops when the substrate binds too far away from the iron and that the phenols bind to microperoxidases at similar distances to those observed with peroxidases. While the reaction rate of microperoxidases with peroxide is significantly smaller than that of the enzymes, the efficiency in the one-electron oxidation of phenolic substrates is almost comparable. Interestingly, the oxyferryl form of the triply positively charged microperoxidases shows a reactivity larger than that exhibited by horseradish peroxidase. PMID- 15532029 TI - Haplotypes of the alpha-1 antitrypsin gene in healthy controls and Z deficiency patients. AB - Alpha-1 antitrypsin (AAT; HUGO symbol, SERPINA1) is one of the major serine protease inhibitors (serpins) in human plasma. Deficiency of AAT is a recognised risk factor for chronic obstructive pulmonary disease (COPD), attributed to uninhibited neutrophil elastase released into the lung tissue during inflammatory states. In this study we used sequencing to screen the exonic regions, 5' and 3' flanking sequence of the AAT gene in order to generate a high density map of single nucleotide polymorphisms (SNPs). 16 SNPs were identified throughout AAT. Haplotypes based on SNPs with a minor allele frequency of > or =5% were estimated using genotypic information from 225 healthy control individuals and 41 AAT deficient Pi-ZZ individuals. AAT shows a large amount of variation in the control population, with 17 haplotypes accounting for 88% of the observed variation. The haplotype distribution of the common deficiency Pi-Z variant of AAT was significantly different when compared to the normal variants. In addition to the haplotype information, we present evidence for a functional effect of a SNP in intron 1. PMID- 15532030 TI - Coexpression of Brn-3a POU protein with p53 in a population of neuronal progenitor cells is associated with differentiation and protection against apoptosis. AB - The Brn-3a transcription factor is critical for survival and differentiation of sensory neurons derived from neural crest cells (NCC). Interaction of Brn-3a with p53 results in differential effects on target gene expression, which profoundly affects fate of neuronal cells. Here we demonstrate colocalization of p53 in a subset of Brn-3a-positive NCC-derived cells fated for the sensory neuronal lineage. The distinct morphology of Brn-3a/p53-coexpressing cells suggested a differentiated neuronal cell type, and this was confirmed by colocalization of p53 with differentiation marker NF-160. Functional effects of Brn-3a/p53 coexpression were analyzed in NCC cultured from Brn-3a -/- embryos, which showed significantly increased apoptosis upon induction of p53 compared with wild-type NCC, suggesting that Brn-3a modulates the p53-mediated fate of NCC that coexpress both factors. Thus, p53 is expressed in neuronal cells undergoing differentiation as well as apoptosis. Interaction with Brn-3a in sensory neurons may be critical for modulating p53-mediated gene expression and hence cell fate. PMID- 15532031 TI - Reorganization of microtubule nucleation during muscle differentiation. AB - Skeletal muscle differentiation involves a complete reorganization of the microtubule network. Nearly 20 years ago, Tassin et al. [1985: J Cell Biol 100:35 46] suggested a mechanism for this reorganization by showing a redistribution of the microtubule organizing center from the centrosome to the nuclear membrane. Little progress has been made since. It is still not clear whether centrosomal proteins are redistributed together, whether microtubules are nucleated at the nuclear membrane or transported there post-nucleation, and whether gamma-tubulin (gammatub) remains necessary for nucleation in myotubes. To investigate these questions, we have examined the redistribution of the centrosomal proteins pericentrin (PC), gammatub, and ninein in the C2 muscle cell line. Immunofluorescence of differentiated myotubes shows PC along the nuclear membrane whereas gammatub is only detected there after pre-fixation detergent extraction. After expression of a GFP-tagged gammatub, we observe a weak fluorescence along the nuclear membrane, confirming the presence of gammatub at a low concentration relative to PC. Microinjection of anti-gammatub antibodies into myotubes blocks microtubule growth from both nuclear membranes and centrosomal sites. The centrosomal microtubule-anchoring protein, ninein, is found at the nuclear membrane as well and its distribution appears independent of microtubule integrity. We conclude that centrosomal proteins are redistributed independently during muscle differentiation, to sites that nucleate microtubules both along the nuclear membranes and through the cytoplasm. PMID- 15532032 TI - A novel mutation in KCNA1 causes episodic ataxia without myokymia. AB - We describe a unique family in which several individual are affected with episodes of ataxia that best fit the phenotype of episodic ataxia type 2 (EA2). All of the affected family members had episodes typically lasting for several hours, and none of them had muscle abnormalities including myokymia. Episodic ataxia type 1 (EA1) was not considered initially as a clinical diagnosis for the affected individuals in this family. However, by linkage mapping, sequencing and polymorphism analysis, all affecteds were found to have a novel mutation in KCNA1. Numerous missense mutations have been described previously in KCNA1 that cause EA1. The mutation c.1025G>T replaces a highly conserved serine with isoleucine at position 342 (p.Ser342Ile) in the highly conserved fifth transmembrane domain of the KCNA1. This mutation leads to a distinct clinical phenotype without myokymia broadening the scope of clinical characteristics of EA1 and highlighting the heterogeneity of phenotypic effects from distinct missense mutations. PMID- 15532033 TI - Accepting its random coil nature allows a partial NMR assignment of the neuronal Tau protein. AB - A combined strategy to obtain a partial NMR assignment of the neuronal Tau protein is presented. Confronted with the extreme spectral degeneracy that the spectrum of this 441 amino acid long unstructured protein presents, we have introduced a graphical procedure based on residue type-specific product planes. Combining this strategy with the search for pairwise motifs, and combining the spectra of different Tau isoforms and even of peptides derived from the native sequence, we arrive at a partial assignment that is sufficient to map the interactions of Tau with its molecular partners. The obtained assignments equally confirm the absence of regular secondary structure in the isolated protein. PMID- 15532034 TI - Hoechst 33258 selectively inhibits group I intron self-splicing by affecting RNA folding. AB - Fungal pathogens are increasing in prevalence due to an increase in resistant strains and the number of immunocompromised humans. Candida albicans is one of these pathogens, and approximately 40% of strains contain a group I self-splicing intron, which is a potential RNA drug target, in their large subunit rRNA precursor. Here, we report that Hoechst 33258 and derivatives thereof are selective inhibitors of C. albicans group I intron self-splicing with an IC50 of 17 microM in 2 mM Mg2+. Chemical probing of the intron in the presence of Hoechst 33258 reveals that the folding of several nucleotides in the P4/P6 region of the intron is affected. A nucleotide near the J4/5 region is protected from chemical modification in the presence of Hoechst 33258 and several nearby are more reactive; this suggests that this region is the molecule's binding site. These results expand the available information on small-molecule targeting of RNA and suggest that the RNA-targeting scaffold provided by Hoechst may prove valuable in designing compounds that inhibit the functions of RNA. PMID- 15532036 TI - Random-effects Cox proportional hazards model: general variance components methods for time-to-event data. AB - Proportional hazards regression models are commonly used to study factors associated with time-to-event data. Because many complex genetic diseases exhibit variation in age at onset, it is important to have the capability to perform survival analyses on data collected from individuals whose observations are correlated due to shared genes or environment. While there are widely accepted methods for variance components analysis for simple quantitative traits, a parallel methodology for survival data has not been available. This manuscript outlines a method to perform variance component analyses under general random effects proportional hazards models. This method is based on a Laplace approximation, and makes computation for correlated time-to-event data feasible. The correlated frailty models described here can be used to perform genetic analyses, and other analyses with structured random effects, on age-at-onset data in a manner analogous to standard variance components methods for quantitative traits. We illustrate the use of the method by examining the heritability of breast cancer in a large familial cohort study. We also perform variance components linkage analyses on data simulated for the Twelfth Genetic Analysis Workshop (GAW12), and further examine the performance of this method for linkage analysis in a simulation study. The breast cancer analyses support significant heritability of disease age-at-onset that is of moderate size. The variance component linkage analyses successfully identify the location of the disease genes that were simulated to have a direct impact on age-at-onset. The methods outlined here make it possible to perform general variance components analyses on time-to-event endpoints, even on large data sets, in a computationally efficient manner. PMID- 15532037 TI - Identifying interacting SNPs using Monte Carlo logic regression. AB - Interactions are frequently at the center of interest in single-nucleotide polymorphism (SNP) association studies. When interacting SNPs are in the same gene or in genes that are close in sequence, such interactions may suggest which haplotypes are associated with a disease. Interactions between unrelated SNPs may suggest genetic pathways. Unfortunately, data sets are often still too small to definitively determine whether interactions between SNPs occur. Also, competing sets of interactions could often be of equal interest. Here we propose Monte Carlo logic regression, an exploratory tool that combines Markov chain Monte Carlo and logic regression, an adaptive regression methodology that attempts to construct predictors as Boolean combinations of binary covariates such as SNPs. The goal of Monte Carlo logic regression is to generate a collection of (interactions of) SNPs that may be associated with a disease outcome, and that warrant further investigation. As such, the models that are fitted in the Markov chain are not combined into a single model, as is often done in Bayesian model averaging procedures. Instead, the most frequently occurring patterns in these models are tabulated. The method is applied to a study of heart disease with 779 participants and 89 SNPs. A simulation study is carried out to investigate the performance of the Monte Carlo logic regression approach. PMID- 15532038 TI - Characterization of the metabolic burden on Escherichia coli DH1 cells imposed by the presence of a plasmid containing a gene therapy sequence. AB - The presence of a plasmid, containing gene sequences for DNA immunotherapy that are not expressed in microbial culture, imposed a degradation in bioreactor performance in cultures of the host E. coli strain. Significant decreases in growth rate (24%) and biomass yield (7%) and a corresponding increase in overflow metabolism were observed in a strain containing a therapeutic sequence (a hepatitis B antigen under the control of a CMV promotor). The observed increase in overflow metabolism was incorporated into a Metabolic Flux Analysis (MFA) model (as acetate secretion). Metabolic flux analysis revealed an increase in TCA cycle flux, consistent with an increased respiration rate observed in plasmid containing cells. These effects are thought to result from increased ATP synthesis requirements (24%) arising from the expression of the Kanr plasmid marker gene whose product accounted for 18% of the cell protein of the plasmid containing strain. These factors will necessitate significantly higher aeration and agitation rates or lower nutrient feed rates in high-density cultures than would be expected for plasmid-free cultures. PMID- 15532039 TI - Thermophilic biohydrogen production from glucose with trickling biofilter. AB - Thermophilic H2 production from glucose was studied at 55-64 degrees C for 234 days using a continuous trickling biofilter reactor (TBR) packed with a fibrous support matrix. Important parameters investigated included pH, temperature, hydraulic retention time (HRT), and glucose concentration in the feed. The optimal pH and temperature were 5.5 and 60 degrees C, respectively. With decreasing HRT or increasing inlet glucose concentration, volumetric H2 production rate increased but the H2 production yield to glucose decreased gradually. The biogas composition was almost constant at 53 +/- 4% (v/v) of H2 and 47 +/- 4% (v/v) of CO2. No appreciable CH4 was detected when the reactor was under a normal operation. The carbon mass balance showed that, in addition to cell mass, lactate, n-butyrate, CO2, and acetate were major products that comprised more than 85% of the carbon consumed. The maximal volumetric H2 production rate and H2 yield to glucose were 1,050 +/- 63 mmol H2/l.d and 1.11 +/ 0.12 mol H2/mol glucose, respectively. These results indicate that the thermophilic TBR is superior to most suspended or immobilized reactor systems reported thus far. This is the first report on continuous H2 production by a thermophilic TBR system. PMID- 15532040 TI - Transient production of recombinant proteins by Chinese hamster ovary cells using polyethyleneimine/DNA complexes in combination with microtubule disrupting anti mitotic agents. AB - We have developed a simple and robust transient expression system utilizing the 25 kDa branched cationic polymer polyethylenimine (PEI) as a vehicle to deliver plasmid DNA into suspension-adapted Chinese hamster ovary cells synchronized in G2/M phase of the cell cycle by anti-mitotic microtubule disrupting agents. The PEI-mediated transfection process was optimized with respect to PEI nitrogen to DNA phosphate molar ratio and the plasmid DNA mass to cell ratio using a reporter construct encoding firefly luciferase. Optimal production of luciferase was observed at a PEI N to DNA P ratio of 10:1 and 5 mug DNA 10(6) cells(-1). To manipulate transgene expression at mitosis, we arrested cells in G2/M phase of the cell cycle using the microtubule depolymerizing agent nocodazole. Using secreted human alkaline phosphatase (SEAP) and enhanced green fluorescent protein (eGFP) as reporters we showed that continued inclusion of nocodazole in cell culture medium significantly increased both transfection efficiency and reporter protein production. In the presence of nocodazole, greater than 90% of cells were eGFP positive 24 h post-transfection and qSEAP was increased almost fivefold, doubling total SEAP production. Under optimal conditions for PEI-mediated transfection, transient production of a recombinant chimeric IgG4 encoded on a single vector was enhanced twofold by nocodazole, a final yield of approximately 5 microg mL(-1) achieved at an initial viable cell density of 1 x 10(6) cells mL( 1). The glycosylation of the recombinant antibody at Asn297 was not significantly affected by nocodazole during transient production by this method. PMID- 15532041 TI - Synergistic effect of shear stress and streptavidin-biotin on the expression of endothelial vasodilator and cytoskeleton genes. AB - Dual ligand treatment of streptavidin(SA)-biotin and fibronectin (Fn) enhances the adhesion of endothelial cells (EC) onto synthetic surfaces and promotes the quiescent phenotype of adherent EC. The current study investigates the effect of the dual ligand on the expression of endothelial genes in static culture and under shear stress (4 h at 10 dynes/cm2). Expression of 23 genes in the classes of signaling, cytoskeleton/ECM, vasoregulation, and shear-responsive were examined. Eight genes (argininosuccinate synthetase, K+ channel, TGFbeta, Mn-SOD, alpha-tubulin, t-PA, COX2, and eNOS) were significantly upregulated by shear stress. Two genes (caveolin-1 and ET-1) were downregulated by shear stress. Three genes (RhoA, elastin, alpha-actinin) were upregulated by the dual ligand treatment in static culture, and four genes (FAK, elastin, COX2, and eNOS) were upregulated when the dual ligand and shear stress were applied simultaneously. Northern blot analyses on FAK, RhoA, elastin, and alpha-actinin revealed similar results. The results suggest (1) the use of SA-biotin to supplement EC adhesion enhances the integrity of the EC cytoskeleton by upregulating the expression of cytoskeleton/ECM genes, and (2) a likely relationship between the expression of cytoskeleton/ECM genes and the downstream events, such as the shear-induced expression of eNOS and COX2 genes. Analyses presented in this study provide insights into the mechanism by which SA-biotin-supplemented EC mediate gene expression. PMID- 15532042 TI - Occurrence of circadian rhythms in hairy root cultures grown under controlled conditions. AB - Hairy roots obtained by transformation via Agrobacterium rhizogenes provide an artificial plant material devoid of aerial parts with high growth on hormone-free media. Fundamental knowledge of hairy root physiology is essential to develop and control its culture. In contrast to shake-flask cultures, a bioreactor set-up combined with on-line data logging provides an efficient tool to study rapid physiological variations in hairy root cultures. Datura innoxia hairy roots were grown in a bioreactor equipped with on-line data analyses of pH, dissolved oxygen (pO2), conductivity, oxygen, and carbon dioxide. The experiments were done at a constant temperature and in the absence of light cues. The results obtained showed that the carbon dioxide evolution rate (CER) presented regular oscillations during the culture. Similar oscillations were also observed for the oxygen uptake rate (OUR). These signals were treated mathematically to look for the existence of a rhythm. An autocorrelation function was used to detect any periodic components. The results demonstrate that hairy root respiration exhibited peaks of 1 day. These oscillations, having a period of about 24 h, were also observed in pH and conductivity signals, although not for the pO2 signal. The data acquired in the absence of hairy roots showed that the observed periodic behavior was not an artifact. No effect on rhythms was observed by the imposition of an external "day/night" cycle. The fact that oscillations persisted in the absence of external stimuli, with a free-running period of 24 h, suggests that a circadian rhythm exists in hairy roots of D. innoxia. PMID- 15532043 TI - Skeletal muscle development in normal and double-muscled cattle. AB - This study examined the effect of genotype on prenatal muscle development in both normal-muscled (NM) animals and in double-muscled (DM) animals harboring a mutation in the gene for myostatin that results in the production of a functionally inactive protein. The following muscle development parameters were analyzed at four gestational ages: muscle weight, fiber type, by both enzyme histochemistry and myosin heavy-chain (MHC) immunocytochemistry, and average fiber area. The weights of both M. vastus lateralis and M. vastus medialis were greater throughout prenatal development in the DM animals compared to NM. The percentage of type 1 muscle fibers initially declined with gestational age and subsequently increased in both NM and DM. The percentage of type 1 fibers was consistently lower in DM than in NM. A pattern of MHC isoform localization was shown in DM muscle that is indicative of a delay in muscle development relative to NM. Muscle fiber size was differentially regulated in NM and DM, depending on fiber type. Type 1 fibers were smaller in DM than NM in late gestation, while type 2 fibers were smaller throughout gestation. This study suggests that the inactivating myostatin mutation in DM animals may be associated with changes in both skeletal muscle fiber type and fiber size during bovine muscle development. PMID- 15532044 TI - Strain improvement and metabolic flux analysis in the wild-type and a mutant Lactobacillus lactis strain for L(+)-lactic acid production. AB - The effects of initial glucose concentration and calcium lactate concentration on the lactic acid production by the parent strain, Lactobacillus lactis BME5-18, were studied. The results of the experiments indicated that glucose and lactate repressed the cell growth and the lactic acid production by Lactobacillus lactis BME5-18. A L(+)-lactic acid overproducing strain, Lactobacillus lactis BME5-18M, was screened by mutagenizing the parent strain with ultraviolet (UV) light irradiation and selecting the high glucose and lactate calcium concentration repression resistant mutant. Starting with a concentration of 100g L(-1) glucose, the mutant produced 98.6 g L(-1) lactic acid after 60 h in flasks, 73.9% higher than that of the parent strain. The L(+)-lactic acid purity was 98.1% by weight based on the amount of total lactic acid. The culture of the parent strain could not be analyzed well by conventional metabolic flux analysis techniques, since some pyruvate were accumulated intracellularly. Therefore, a revised flux analysis method was proposed by introducing intracellular pyruvate pool. Further studies demonstrate that there is a high level of NADH oxidase activity (12.11 mmol mg(-1) min(-1)) in the parent strain. The molecular mechanisms of the strain improvement were proposed, i.e., the high level of NADH oxidase activity was eliminated and the uptake rate of glucose was increased from 82.1 C-mmol (g DW h)(-1) to 98.9 C-mmol (g DW h)(-1) by mutagenizing the parent strain with UV, and therefore the mutant strain converts mostly pyruvate to lactic acid with a higher productivity (1.76 g L(-1) h(-1)) than the parent strain (0.95 g L(-1) h(-1)). PMID- 15532046 TI - Significant glomerular basement membrane thickening in hyperglycemic and normoglycemic diabetic-prone BB Wistar rats. AB - The diabetic-prone BioBreeding Wistar rat (BB/DP) is an autoimmune model of insulin-dependent diabetes mellitus. Approximately 80-90% of the animals are hyperglycemic (BB/DP(h)) by 90-120 days of age while those that do not become diabetic in adolescence (BB/DP(n)) remain normoglycemic for life. Likewise, rats in the diabetes-resistant (BB/DR) strain are normoglycemic. Although renal morphological studies have been carried out in this model, ultrastructural observations of age- and diabetes-related extracellular matrix (ECM) changes, including glomerular basement membrane (GBM) morphometry, are not available. Moreover, possible renal changes in the relatively uncommon BB/DP(n) control animals have not been reported. The current electron microscopic study was carried out to investigate temporal changes in detergent-treated acellular ECM in BB/DP(h) rats at 2 weeks, 3 months, 6 months, and 1 year postonset of moderate hyperglycemia. Age-matched BB/DR and BB/DP(n) control animals were also examined. Our data demonstrate age- and diabetes-related alterations in mesangial matrix distributions and GBM widths and show for the first time significant increases in GBM thickening in both hyperglycemic (BB/DP(h)) and normoglycemic (BB/DP(n)) rats when compared to age-matched BB/DR controls. Surprisingly, the rate of increase is greatest in BB/DP(n) animals. Although the pathogenesis of diabetic basement membrane disease is not completely understood, GBM thickening is widely regarded as a morphological consequence of hyperglycemia. However, data in the current investigation show that ECM alterations, including significantly increased GBM thickness, may occur in genetically diabetic animals in the absence of hyperglycemia. PMID- 15532045 TI - Identification, quantification and comparison of major non-polar lipids in normal and dry eye tear lipidomes by electrospray tandem mass spectrometry. AB - Millions of individuals suffer from a health condition known as keratoconjunctivitis sicca (KCS, also known as 'dry eye'). Studies have indicated that the lipids in the tear film layer, which covers the outer portion of the eye, may be directly correlated with the existence of dry eye syndrome. By identifying and comparing the major, non-polar lipids in normal eye tears with a dry eye model, it may be possible to identify a symptom of, or a contributing factor to, dry eye. Electrospray tandem mass spectrometry (ES-MS/MS) was used to identify and compare the non-polar lipids, detected as lithium adducts, from normal and dry eye tear samples obtained from rabbits. A limited number of normal human tear samples were also examined for lipid content, and a close resemblance to rabbit was observed. Three distinct regions were delineated in the ES mass spectra of the non polar lipids, m/z 20-500, 500-800 and 800-1100. A common feature noted among identified lipid components was a glycerol backbone with fatty acyl substituents attached. Product ion spectra were obtained for lithiated monoacyl-, 1,2- and 1,3-diacyl- and triacylglyceride standards. Newly proposed structures and fragmentation pathways for the major product ions are presented for the 1,2- and 1,3-diglycerides, and also for the monoglyceride. New approaches to distinguishing asymmetric 1,2-diglycerides and 1,2- from 1,3-diglycerides are proposed. For the rabbit tear samples, the m/z 20-500 range contains monoester diols with empirical formulas C(n)H(2n)O(4), the m/z 500-800 range includes diesters with empirical formulas C(n)H(2n-2)O(5) and the m/z 800-1100 range contains triesters with empirical formulas C(n)H(2n-4)O(6). Also found in the extracts were three isoprene acetals (terpenoids). PMID- 15532047 TI - Angiogenic response induced by acellular aortic matrix in vivo. AB - In this study, we investigated the angiogenic response induced by acellular aortic matrices implanted in vivo onto the chick embryo chorioallantoic membrane (CAM), a useful model for such investigation. Results showed that acellular matrices were able to induce a strong angiogenic response comparable to that of fibroblast growth factor 2 (FGF-2), a well-known angiogenic cytokine. The angiogenic response was further increased when exogenous FGF-2 or transforming growth factor beta 1 (TGF-beta1) were added to the matrices and inhibited by the addition of an anti-FGF-2 or anti-TGF-beta1 antibodies. The response may be considered dependent on a direct angiogenic effect exerted by the matrices and in part also by the presence of FGF-2 and TGF-beta1 in the acellular matrices. PMID- 15532048 TI - Fast detection of single nucleotide polymorphisms (SNPs) by primer elongation with monitoring of supercritical-angle fluorescence. AB - We describe the rapid detection of single nucleotide polymorphisms (SNPs) by real time observation of primer elongation. The enzymatic elongation of surface-bound primers is monitored by detecting the increase of surface-bound fluorescence caused by the incorporation of Cy5-labelled deoxycytidine 5'-triphosphate residues (Cy5-dCTPs) into the corresponding strand. In order to discriminate against the fluorescence from unbound Cy5-dCTPs, the detection volume was restricted to the surface by collecting supercritical-angle fluorescence. The efficiency of enzymatic double-stranded DNA synthesis is governed by the complementarity of the primer and template. An SNP in the sequence of the primer obstructs its elongation increasingly with decreased distance of the mismatch to the 3' end of the primer. By real-time fluorescence detection during primer elongation, SNPs can be detected within a few minutes, which is significantly faster than in experiments where the fluorescence is measured after completion of the reaction. We demonstrate the efficiency of the method by detecting an SNP in the ErbB2 gene that is involved in causing a higher risk of breast cancer. PMID- 15532049 TI - Novel formulations of vitamins and insulin by nanoengineering of polyelectrolyte multilayers around microcrystals. AB - Microcapsules loaded with vitamin K3 (VK3), biotin, or insulin were prepared by using a novel coating technology based on the layer-by-layer (LbL) deposition of oppositely charged polyelectrolytes onto microcrystal templates. This produced multilayered, polymeric shells of varying thickness around the crystalline cores. Dissolution of the core material (VK3 with ethanol, biotin with basic solution, and insulin with acidic solution), resulted in its release through the shells. Microelectrophoresis was employed to monitor the microcrystal coating process; confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) were used to verify multilayer coating and the formation of hollow polymer shells following removal of the microcrystal templates. The release rates of both VK3 and insulin decreased as the wall thickness (the number of polyelectrolyte layers deposited onto the microcrystal cores), increased. The release time could be varied by a factor of more than ten, depending on the number of polyelectrolyte layers applied. Following the addition of 70 mass % ethanol, the solubility of VK3 increased by as much as 170-fold, resulting in an increased rate of VK3 release. By selecting appropriate polymer materials for the shells, and by controlling the number of polyelectrolyte layers applied, shells of various thickness, stiffness, aqueous solubility, dispersibility, biocompatibility, and permeability can be constructed. PMID- 15532050 TI - Role of the special pair in the charge-separating event in photosynthesis. AB - We synthesized special-pair/electron-acceptor systems consisting of a complementary slipped cofacial dimer of imidazolyl-substituted zinc porphyrin, bearing pyromellitdiimide as the electron acceptor. In the case of the dimer, the first and second oxidation potentials were split into a total of four peaks in the differential pulse voltammetry measurement. Furthermore, the shift values of the first oxidation potentials obtained by changing the solvent polarity for the dimer were almost half of those observed for the monomer. These results indicate that the radical cation is delocalized over the whole pi system of the dimer. Time-resolved transient absorption measurements revealed that, relative to the corresponding monomer, the dimer accelerated the charge separation rate, but decelerated the charge recombination rate. The smaller reorganization energy of the slipped cofacial dimer relative to that of the monomeric system demonstrates the significance of the special-pair arrangement for efficient charge separation in photosynthesis. PMID- 15532051 TI - Intramolecular rearrangement for regioselective complexation by intramolecular CH/pi interaction in a hydrophobic cavity of a ruthenium coordination sphere. AB - A Ru(II) complex with a hydrophobic cavity formed from two 1-naphthoylamide groups was prepared. Its reactions with beta-diketones gave beta-diketonato complexes in which hydrophobic pi-pi or CH/pi interactions were confirmed by NMR spectroscopy and X-ray crystallography. In the case of the asymmetric beta diketone benzoylacetone, an isomer with a CH/pi interaction was afforded as the sole product owing to thermodynamic control. The reaction was found to involve a novel intramolecular rearrangement from the phenyl-included isomer to the methyl included one without rupture of Ru-beta-diketonato coordination bonds (activation energy 52 kJ mol(-1)). This indicates that CH/pi interactions can be more favored thermodynamically than pi-pi interactions in a suitable hydrophobic environment. PMID- 15532052 TI - Preparation and redox properties of N,N,N-1,3,5-trialkylated flavin derivatives and their activity as redox catalysts. AB - Eight different flavin derivatives have been synthesized and the electronic effects of substituents in various positions on the flavin redox chemistry were investigated. The redox potentials of the flavins, determined by cyclic voltammetry, correlated with their efficiency as catalysts in the H2O2 oxidation of methyl p-tolyl sulfide. Introduction of electron-withdrawing groups increased the stability of the reduced catalyst precursor. PMID- 15532053 TI - Isotope exchange in ionised CO2/CO mixtures: the role of asymmetrical C2O3+ ions. AB - A hitherto unknown, atmospherically relevant, isotope-exchange reaction was studied in ionised gaseous mixtures containing carbon dioxide and monoxide. The mechanism of the O exchange, proceeding over a double-minimum potential-energy surface, was positively established by mass spectrometric and theoretical methods that also allowed the identification and characterisation of the C2O3+ intermediate. The increase of internal energy displaces the observed reactivity towards an endothermic reaction path that involves only CO2 and represents an indirect route to the dissociation of carbon dioxide. PMID- 15532054 TI - The mechanism of magnetic interactions in the bulk ferromagnet para (methylthio)phenyl nitronyl nitroxide (YUJNEW): a first principles, bottom-up, theoretical study. AB - The mechanism of magnetic interactions in the bulk ferromagnet para (methylthio)phenyl nitronyl nitroxide crystal (YUJNEW) has been theoretically reinvestigated, using only data from ab initio calculations and avoiding any a priori assumptions. We first calculate the microscopic magnetic interactions (JAB exchange couplings) between all unique radical pairs in the crystal, and then generate the macroscopic magnetic properties from the energy levels of the corresponding Heisenberg Hamiltonian. We thus propose a first principles, bottom up (i.e. micro-to-macro) approach that brings theory and experiment together. We have applied this strategy to study the magnetism of YUJNEW using data from the previously reported 298 and 114 K crystal structures, and also data from a 10 K neutron diffraction structure fully reported in this work. The magnetic topology at 298 K is two-dimensional: noninteracting planes, with three different in-plane JAB pair interactions (+0.24, +0.09, and -0.11 cm(-1)) and one numerically negligible (+0.02 cm(-1)) inter-plane JAB interaction. In contrast, the magnetic topology at 114 and 10 K is three-dimensional, with two non-negligible in-plane JAB constants (+0.11 and +0.07 cm(-1) at 114 K; +0.22 and +0.07 cm(-1) at 10 K) and one inter-plane pair interaction (+0.07 cm(-1) at 114 K; +0.08 cm(-1) at 10 K). Although this three-dimensional magnetic topology is consistent with YUJNEW being a bulk ferromagnet, there is only a qualitative agreement between computed and experimental magnetic susceptibility chiT(T) data at 114 K. However, the experimental chiT(T) curve is quantitatively reproduced at 10 K. The heat capacity curve presents a peak at around 0.12 K, close to the estimated experimental peak (0.20 K). PMID- 15532055 TI - Self-assembly of folic acid derivatives: induction of supramolecular chirality by hierarchical chiral structures. AB - Hierarchical chiral structures made up of dendritic oligo(L- or D-glutamic acid) moieties of folic acid derivatives induce supramolecular chirality in the self assembled columnar structures of the folic acids. These folic acids self-assemble through the intermolecular hydrogen bonds of the pterin rings to form disklike tetramers. In the neat states, the stacked tetramers form thermotropic hexagonal columnar phases over wide temperature ranges, including room temperature. Addition of alkali metal salts induces chirality in the columnar phases. In dilute solution states in a relatively polar solvent (chloroform), the folic acid derivatives form non-chiral, self-assembled structures. In the presence of sodium triflate, the folic acid forms chiral columnar assemblies through the oligo(L glutamic acid) moiety, similar to those formed in the liquid-crystalline (LC) states. The enantiomer of the folic acid induces columnar assemblies with reversed helicity. In the case of the diastereomer, no induced helicity is observed. Application of an apolar solvent (dodecane) drives the folic acid derivatives to form chiral assemblies in the absence of ions. In this case, lipophilic interactions promote nanophase segregation, which enhances the formation of chiral columns. Interestingly, the chiral supramolecular structure of the diastereomer induces the most intense circular dichroism. In both cases, the molecular chirality in the oligo(glutamate) moieties yields supramolecular chirality of the folic acids that self-assemble through cooperative molecular interactions. PMID- 15532056 TI - Palladium-catalyzed intramolecular oxidative alkylation of 4-pentenyl beta dicarbonyl compounds. AB - Reaction of 8-nonene-2,4-dione with a catalytic amount of [PdCl2(CH3CN)2] (2; 5 mol %) and a stoichiometric amount of CuCl2 (2.5 equiv) at room temperature for 3 h led to oxidative alkylation and formation of 2-acetyl-3-methyl-2-cyclohexenone in 80 % isolated yield. The oxidative alkylation of 4-pentenyl beta-diketones tolerated a number of terminal acyl groups and substitution at the C1 and C3 carbon atoms of the 4-pentenyl chain. Likewise, 4-pentenyl beta-keto esters that possessed geminal disubstitution at the C1, C2, or C3 carbon atom of the 4 pentenyl chain cyclized to form 2-carboalkoxy-2-cyclohexenones in moderate to good yield as the exclusive cyclized product. Deuterium-labeling experiments provided information regarding the mechanism of the palladium-catalyzed oxidative alkylation of 4-pentenyl beta-dicarbonyl compounds. PMID- 15532057 TI - FGF-2 signaling is sufficient to induce dermal condensations during feather development. AB - In a previous report, we showed that fibroblast growth factor-2 (FGF-2) is a signal produced by epidermal placode cells during feather development and that this growth factor can induce feathers in scaleless mutant skins that fail to form feathers due to a defective epidermis (Song et al., [1996] Proc Natl Acad Sci USA 93:10246-10249). Here, we test whether FGF-2 is sufficient to induce dermal condensations, structures that normally form under the control of signals from the epidermal placode and are identified on the basis of aggregation of cells, the expression of FGF receptor-1 and bone morphogenetic protein-2 transcripts and the cessation of proliferation of the condensed cells. By using denuded 8-day scaleless dermis as a test system, we have established that FGF-2 is sufficient to induce dermal condensation. We suggest that the primary effect of FGF-2 is an increase in cellular density mediated through cell migration, followed by the expression of dermal condensation-specific genes and cessation of cell proliferation. The FGF-2 effect can be abolished by heparin, suggesting the involvement of heparan sulfate proteoglycans (HSPGs) in growth factor signaling. The spatiotemporal expression of syndecan-3 during feather development suggests that this cell-surface HSPG may be involved in the response of competent embryonic skin dermis to FGF-2. PMID- 15532058 TI - Fgf signaling induces posterior neuroectoderm independently of Bmp signaling inhibition. AB - Whereas according to the neural default model, neural specification is induced by extracellular inhibitors of bone morphogenetic proteins (Bmps), the role of fibroblast growth factors (Fgfs) during neural induction is heavily debated. Here, we show that, in zebrafish embryos, Bmps and Fgfs play differential roles during the induction and patterning of the anterior vs. the posterior neuroectoderm. Induction of anterior neuroectoderm, giving rise to fore- and midbrain, is accomplished by Bmp inhibition, with Fgfs playing a moderate posteriorizing/patterning role, possibly by blocking Bmp signaling at the level of Smad proteins. In contrast, in the posterior-most neuroectoderm, which is located in marginal regions of the early gastrula embryo to give rise to spinal cord and hindbrain, Fgfs play a neural-inducing rather than a neural-patterning role. This Fgf-dependent posterior neural induction takes place during late blastula and early gastrula stages, after mesoderm has been induced and cannot be blocked by Bmps or the Bmp target gene and downstream effector Delta Np63 alpha, indicating that here, Fgfs act independently of Bmp signaling inhibition. PMID- 15532059 TI - Increased megakaryopoiesis in cultures of CD34-enriched cord blood cells maintained at 39 degrees C. AB - Based on previous evidence suggesting positive effects of fever on in vivo hematopoiesis, we tested the effect of hyperthermia on megakaryopoiesis (MK) in ex vivo cultures of CD34-enriched cord blood (CB) cells. The cells were cultured at 37 degrees C or 39 degrees C for 14 days in cytokine conditions optimized for megakaryocyte development and analyzed periodically. Compared to 37 degrees C, the cultures maintained at 39 degrees C produced significantly more (up to 10 fold) total cells, myeloid and MK progenitors, and total MKs, and showed accelerated and enhanced MK maturation with increased yields of proplatelets and platelets. This observation could facilitate clinical applications requiring ex vivo expansion of hematopoietic cells. PMID- 15532060 TI - DNA codes for nanoscience. AB - The nanometer scale is a special place where all sciences meet and develop a particularly strong interdisciplinarity. While biology is a source of inspiration for nanoscientists, chemistry has a central role in turning inspirations and methods from biological systems to nanotechnological use. DNA is the biological molecule by which nanoscience and nanotechnology is mostly fascinated. Nature uses DNA not only as a repository of the genetic information, but also as a controller of the expression of the genes it contains. Thus, there are codes embedded in the DNA sequence that serve to control recognition processes on the atomic scale, such as the base pairing, and others that control processes taking place on the nanoscale. From the chemical point of view, DNA is the supramolecular building block with the highest informational content. Nanoscience has therefore the opportunity of using DNA molecules to increase the level of complexity and efficiency in self-assembling and self-directing processes. PMID- 15532061 TI - Screening for improved cell performance: selection of subclones with altered production kinetics or improved stability by cell sorting. AB - One of the major problems in the biotechnology industry is the selection of cell lines well suited for production of biopharmaceutical proteins. Usually, the most important selection criterion is the cell specific production rate. Nevertheless, a good producer cell line should have a number of additional advantageous properties, which allow the cell line to perform well in the type of bioreactor chosen for the process. However, the time and work required to select for high production rates as well as the lack of methods to specifically select for other cellular properties, usually prevents researchers from including such criteria into their screening program. With the Single Cell Secretion Assay it is possible to measure the specific production rates of individual cells by catching secreted product in an artificial matrix applied to the cell surface. Flow cytometric cell sorting then allows selection of rare cells with high production rates, which occur at frequencies as low as 10(-6). By combining this method with culture conditions that bring out a desired cellular property, we were able to isolate subclones with similar production rates, but improved performance from a recombinant Chinese hamster ovary cell line producing a human monoclonal antibody. The two desired cellular properties screened for were a non-growth associated production kinetic and improved stability in the absence of selective pressure. PMID- 15532062 TI - Synthesis of isomaltooligosaccharides and oligodextrans in a recycle membrane bioreactor by the combined use of dextransucrase and dextranase. AB - A recycle ultrafiltration membrane reactor was used to develop a continuous synthesis process for the production of isomaltooligosaccharides (IMO) from sucrose, using the enzymes dextransucrase and dextranase. A variety of membranes were tested and the parameters affecting reactor stability, productivity, and product molecular weight distribution were investigated. Enzyme inactivation in the reactor was reduced with the use of a non-ionic surfactant but its use had severe adverse effects on the membrane pore size and porosity. During continuous isomaltooligosaccharide synthesis, dextransucrase inactivation was shown to occur as a result of the dextranase activity and it was dependent mainly on the substrate availability in the reactor and the hydrolytic activity of dextranase. Substrate and dextranase concentrations (50-200 mg/mL(-1) and 10-30 U/mL(-1), respectively) affected permeate fluxes, reactor productivity, and product average molecular weight. The oligodextrans and isomaltooligosaccharides formed had molecular weights lower than in batch synthesis reactions but they largely consisted of oligosaccharides with a degree of polymerization (DP) greater than 5, depending on the synthesis conditions. No significant rejection of the sugars formed was shown by the membranes and permeate flux was dependent on tangential flow velocity. PMID- 15532063 TI - Pulsed electric field stimulates plant secondary metabolism in suspension cultures of Taxus chinensis. AB - The effects of pulsed electric field (PEF) on growth and secondary metabolite production by plant cell culture were investigated by using suspension cultures of Taxus chinensis as a model system. Cultured cells in different growth phases were exposed to a PEF (50 Hz, 10 V/m) for various periods of time. A significant increase in intracellular accumulation of taxuyunnanine C (Tc), a bioactive secondary metabolite, was observed by exposing the cells in the early exponential growth phase to a 30-min PEF. The Tc content (i.e., the specific production based on dry cell weight) was increased by 30% after exposure to PEF, without loss of biomass, compared with the control. The combination of PEF treatment and sucrose feeding proved useful for improving secondary metabolite formation. Production levels of reactive oxygen species, extracellular Tc, and phenolics were all increased, whereas cell capacitance was decreased with PEF treatment. The results show that PEF induced a defense response of plant cells and may have altered the cell/membrane's dielectric properties. PEF, an external stimulus or stress, is proposed as a promising new abiotic elicitor for stimulating secondary metabolite biosynthesis in plant cell cultures. PMID- 15532064 TI - Lignans in resin of Araucaria angustifolia by gas chromatography/mass spectrometry. AB - Total extract of resin from Araucaria angustifolia was analyzed by gas chromatography/mass spectrometry and 32 lignans were identified. Lignan acetates are present in the resin and consist of four secoisolariciresinol acetates, six lariciresinol acetates, two 7'-hydroxylariciresinol acetates and an isolariciresinol acetate, which have hitherto not been reported in the plant kingdom. Shonanin and 7'-hydroxylariciresinol type lignans are also present in A. angustifolia resin. Lignans containing syringyl moieties, characteristic for angiosperms, occur in the resin and consist of 5-methoxylariciresinol-9-acetate, 5'-methoxylariciresinol-9-acetate, 5-methoxypinoresinol dimethyl ether and 5 methoxypinoresinol. This is noteworthy because syringyl moieties have only been reported for Thuja species (Cupressaceae) among the gymnosperms. The mass spectra of the various lignan trimethylsilyl derivatives are discussed with the interpretations of the fragmentation patterns. PMID- 15532065 TI - Locked nucleic acids and intercalating nucleic acids in the design of easily denaturing nucleic acids: thermal stability studies. AB - Intercalating nucleic acids (INA(R)s) with insertions of (R)-1-O-(1 pyrenylmethyl)glycerol were hybridized with locked nucleic acids (LNAs). INA/LNA duplexes were found to be less stable than the corresponding DNA/LNA duplexes when the INA monomer was inserted as a bulge close to the LNA monomers in the opposite strand. This property was used to make "quenched" complements that possess LNA in hairpins and in duplexes and are consequently more accessible for targeting native DNA. The duplex between a fully modified 13-mer LNA sequence and a complementary INA with six pyrene residues inserted after every second base as a bulge was found to be very unstable (Tm=30.1 degrees C) in comparison with the unmodified double-stranded DNA (Tm=48.7 degrees C) and the corresponding duplexes of LNA/DNA (Tm=81.6 degrees C) and INA/DNA (Tm=66.4 degrees C). A thermal melting experiment of a mixture of an LNA hairpin, with five LNA nucleotides in the stem, and its complementary DNA sequence gave a transition with an extremely low increase in optical density (hyperchromicity). When two INA monomers were inserted into the stem of the LNA hairpin, the same experiment resulted in a significant hyperchromicity comparable with the one obtained for the corresponding DNA/DNA duplex. PMID- 15532066 TI - Synthesis, inhibition properties, and theoretical study of the new nanomolar trehalase inhibitor 1-thiatrehazolin: towards a structural understanding of trehazolin inhibition. AB - A new trehazolin analogue, 1-thiatrehazolin, has been synthesized from carbohydrate precursors by a highly efficient route based on our previously developed ketone/oxime ether reductive carbocyclization reaction for the construction of the cyclitol ring and an intramolecular nucleophilic displacement reaction for the construction of the thiazoline ring. 1-Thiatrehazolin is a very potent, slow, tight-binding trehalase inhibitor. A structural model for trehalase inhibition by trehazolin and its analogues, based on the experimental results and supported by theoretical calculations, is proposed. PMID- 15532067 TI - Determination of bromazepam in human plasma by high-performance liquid chromatography with electrospray ionization tandem mass spectrometric detection: application to a bioequivalence study. AB - A simple method using a one-step liquid-liquid extraction (LLE) followed by high performance liquid chromatography (HPLC) with positive ion electrospray ionization tandem mass spectrometric (ESI-MS/MS) detection was developed for the determination of bromazepam in human plasma, using lorazepam as internal standard. The acquisition was performed in the multiple reaction monitoring mode, monitoring the transitions: m/z 316 > 182 for bromazepam and m/z 321 > 275 for lorazepam. The method was linear over the studied range (1-100 ng ml(-1)), with r(2) > 0.98, and the run time was 2.5 min. The intra- and inter-assay precisions were 2.7-14.6 and 4.1-17.3%, respectively and the intra- and inter-assay accuracies were 87-111 and 75.8-109.5%, respectively. The mean recovery was 73.7%, ranging from 64.5 to 79.7%. The limit of quantification was 1 ng ml(-1). At this concentration the mean intra- and inter-assay precisions were 14.6 and 7.1%, respectively, and the mean intra- and inter-assay accuracies were 102.5 and 104%, respectively. Bromazepam stability was evaluated and the results showed that the drug is stable in standard solution and in plasma samples under typical storage and processing conditions. The method was applied to a bioequivalence study in which 27 healthy adult volunteers (14 men) received single oral doses (6 mg) of reference and test bromazepam formulations, in an open, two-period, randomized, crossover protocol. The 90% confidence interval of the individual ratios (test formulation/reference formulation) for C(max) (peak plasma concentration), AUC(0-96) and AUC(0-inf) (area under the plasma concentration versus time curve from time zero to 96 h and to infinity, respectively) were within the range 80-125%, which supports the conclusion that the test formulation is bioequivalent to the reference formulation regarding the rate and extent of bromazepam absorption. PMID- 15532068 TI - Relationship between local structural entropy and protein thermostability. AB - We developed a technique to compute structural entropy directly from protein sequences. We explored the possibility of using structural entropy to identify residues involved in thermal stabilization of various protein families. Examples include methanococcal adenylate kinase, Ribonuclease HI and holocytochrome c(551). Our results show that the positions of the largest structural entropy differences between wild type and mutant usually coincide with the residues relevant to thermostability. We also observed a good linear relationship between the average structural entropy and the melting temperatures for adenylate kinase and its chimeric constructs. To validate this linear relationship, we compiled a large dataset comprised of 1153 sequences and found that most protein families still display similar linear relationships. Our results suggest that the multitude of interactions involved in thermal stabilization may be generalized into the tendency of proteins to maintain local structural conservation. The linear relationship between structural entropy and protein thermostability should be useful in the study of protein thermal stabilization. PMID- 15532069 TI - Structural characterization of 5,6-dichlorotetradecanoic acid, an isolated metabolite of 9,10-dichlorooctadecanoic acid, by studying picolinyl esters, pyrrolidides and methyl esters with electron ionization mass spectrometry. AB - A method is presented for identification of positional isomers of dichlorinated fatty acids, based on derivatization to picolinyl esters prior to gas chromatographic/mass spectrometric analysis in the electron ionization mode. The mass spectra of the picolinyl esters showed structure-specific fragmentation patterns. By using the picolinyl ester, 5,6-dichlorotetradecanoic acid was identified as a metabolite from a cell-culture medium obtained by culturing human cell lines in media supplemented with threo-9,10-dichlorooctadecanoic acid. This indicates that dichlorinated fatty acids are degraded by beta-oxidation. It is also possible to locate tentatively the position of chlorine atoms in 5,6 dichlorotetradecanoic acid as its methyl ester or pyrrolidide. PMID- 15532070 TI - On-line size-exclusion chromatography/mass spectrometry of low molecular mass heparin. AB - Heparin and low molecular mass heparin (LMMH) consists of complex mixtures of sulphated linear oligosaccharides that are difficult to analyse. An on-line size exclusion chromatographic/electrospray ionization (ESI) mass spectrometric method that allows the determination of more than 60 components in an LMMH preparation is presented. The experimental setup includes on-line cation exchange in order to prevent massive adducting in the ESI interface. PMID- 15532071 TI - Current literature in mass spectrometry. PMID- 15532073 TI - Mass spectrometric analysis of 7-sulfoxymethyl-12-methylbenz[a]anthracene and related electrophilic polycyclic aromatic hydrocarbon metabolites. AB - The Meso-region theory of polycyclic aromatic hydrocarbon (PAH) carcinogenesis predicts that the development of pronounced carcinogenicity depends on the introduction of a good leaving group on alkyl side-chains attached to the exceptionally reactive meso-anthracenic or L-region positions of PAHs. Thus, the first step in carcinogenesis by methylated PAHs such as 7,12 dimethylbenz[a]anthracene (DMBA) would be the hydroxylation of the L-region methyl groups, particularly the 7-methyl group. The second would be the formation of a metabolite, e.g. a sulfate ester, which is expected to be a good leaving group capable of generating a highly reactive benzylic carbocation. 7 Hydroxymethyl-12-methylbenz[a]anthracene (7-HMBA) is a metabolite of DMBA, and sulfation of 7-HMBA to a 7-sulfoxymethyl metabolite (7-SMBA) is a known Phase II metabolic process designed to facilitate excretion, but actually enabling more destructive side-reactions. These side-reactions occur with generation of an electrophilic 7-methylene carbonium ion, and/or by in vivo halide exchange to provide neutral side-products more capable of entering cells, especially those of DMBA target tissues. Electrospray ionization mass spectrometry (MS) enabled us to visualize 7-SMBA as an intact m/z 351 conjugate anion by negative mode, and as a released m/z 255 carbonium ion by positive mode. Upon prolonged refrigeration, 7 SMBA accumulated an m/z 383 photooxide, which appeared capable of re-evolving the starting material as visualized by tandem quadrupole MS, or MS/MS. The 7-SMBA carbonium ion provided interpretable fragments when studied by fragment ion MS/MS, including those representing the loss of up to several protons. Subtle differences in this property were encountered upon perturbing 7-SMBA, either by warming it at 37 degrees C for 2 h or by substituting the initial sulfoxy group with an iodo group. Side-reactions accounting for such proton losses are proposed, and are of interest whether they occur in the mass spectrometer, in solution or both; these proposals include acidity at the 12-methyl position and cyclization between the 12-methyl group and the adjacent C-1 position. It is also suggested that such side-reactions may comprise one route to relieving steric strain arising between the 12-methyl group and the angular benzo ring of 7-SMBA. PMID- 15532075 TI - Structural elucidation and identification of alkaloids in Rhizoma Coptidis by electrospray ionization tandem mass spectrometry. AB - Simple, convenient, sensitive and accurate analytical methods are needed for the structural characterization and identification of alkaloid components in Rhizoma Coptidis in traditional Chinese herbal medicine, which has important bioactivity. In this work, the identification of alkaloid compounds in Rhizoma Coptidis was investigated by obtaining molecular mass information using electrospray ionization mass spectrometry (ESI-MS). Multi-stage tandem mass spectrometric (ESI MS(n)) data for the alkaloid compounds were used for detailed structural characterization, then structure information was obtained by comparison of the fragmentation mechanisms of both alkaloids in Rhizoma Coptidis and standard samples of berberine, palmatine, coptisine and jatrorrhizine by MS. Based on the results obtained, the structure of a novel compound was elucidated. The results of the experiments demonstrate that ESI-MS(n) is a sensitive, selective and effective tool for the rapid determination of alkaloids in Rhizoma Coptidis. PMID- 15532076 TI - Bayesian inference on the patient population size given list mismatches. AB - In applying capture-recapture methods for closed populations to epidemiology, one needs to estimate the total number of people with a certain disease in a certain research area by using several lists with information of patients. Problems of lists error often arise due to mistyping or misinformation. Adopting the concept of tag-loss methodology in animal populations, Seber et al. (Biometrics 2000; 56:1227-1232) proposed solutions to a two-list problem. This article reports an interesting simulation study, where Bayesian point estimates based on improper constant and Jeffreys prior for unknown population size N could have smaller frequentist standard errors and MSEs compared to the estimates proposed in Seber et al. (2000). The Bayesian credible intervals based on the same priors also have super frequentist coverage probabilities while some of the frequentist confidence intervals procedures have drastically poor coverage. Seber's real data set on gestational diabetics is analysed with the proposed new methods. PMID- 15532077 TI - Re: "What is the role of tests of lung function in the management of infants with lung disease?" by Godfrey et al. (Pediatr Pulmonol 2003;36:1-9): interpretation and application of infant lung function tests. PMID- 15532078 TI - Presentation and treatment of asthma among native children in southwest Alaska delta. AB - Our objective was to determine if a different presentation of asthma among Eskimo children in southwest Alaska influenced treatment for asthma. Data regarding symptoms, medication use, and hospitalization were obtained from the medical records of 58 Eskimo children diagnosed with asthma. Half of the children also had a diagnosis of chronic lung disease (CLD), and 57% had a history of allergies. CLD was associated with significantly more visits for wheeze (P=0.02), asthma (P <0.005), and lower respiratory tract illnesses (P <0.005), and a greater incidence (P <0.005) and frequency (P <0.005) of hospitalizations. Allergy status showed no similar relation with utilization of health services. Inhaled corticosteroids were prescribed for a minority (38%) of these asthmatic children. Allergic children tended to be more likely to receive inhaled steroids, and they received significantly more prescriptions for inhaled steroids compared to children without allergies. Those with CLD only were no more likely to receive inhaled steroids than other children, despite their higher incidence of hospitalization. Although the proportion of children with CLD or allergy did not differ significantly by village, there were significant regional differences in healthcare utilization and medication use. In conclusion, while CLD was the primary determinant of healthcare utilization among these native children with asthma, only allergic children with CLD were more likely to receive inhaled steroids. PMID- 15532079 TI - DNA concentrations in BAL fluid of cystic fibrosis patients with early lung disease: influence of treatment with dornase alpha. AB - Recombinant DNase (dornase alpha) was shown to improve lung function and reduce pulmonary exacerbations in cystic fibrosis (CF) patients, but its effects on DNA concentrations in the lower airways remain unclear at the present time. As part of the Bronchoalveolar Lavage in the Evaluation of Anti-Inflammatory Treatment (BEAT) Study, a multicenter open study to evaluate the evolution of inflammation in CF patients with early lung disease and its modulation by dornase alpha treatment, we studied DNA concentrations in the bronchoalveolar lavage (BAL) fluid of 48 CF patients with mild lung disease. After the initial BAL, 29 patients received daily treatment with 2.5 mg of dornase alpha; 19 patients served as controls. BAL was repeated after 18 months in all patients. Mean BAL fluid DNA concentrations were not different between groups at baseline (mean +/- SD, 14.1 +/- 6.9 microg/ml for controls, and 17.6 +/- 11.2 microg/ml for the dornase alpha group), but higher than previously reported for infants with CF. A weak but positive correlation (P <0.01) was observed between the percentage of neutrophils in BAL fluid and DNA levels. On reassessment after 18 months, the percentage of neutrophils was not different between the two groups, but DNA had increased in controls, whereas decreased levels were observed in treated patients (P <0.03, t-test). DNA concentrations increased by more than 10 microg/ml in 7 of 19 controls compared to 2 of 29 CF patients treated with rhDNase (P=0.01, Fisher's test). Therefore, treatment with dornase alpha over 18 months reduces DNA load in BAL fluid, which may have a positive effect on the clearance of lower airway secretions. PMID- 15532080 TI - Trampoline use as physiotherapy for cystic fibrosis patients. AB - Physicians and physiotherapists who care for CF patients have recommended the use of trampolines as a physiotherapeutic tool for enhancing cardiopulmonary performance, encouraging sputum production, and improving general well-being. Despite some therapeutic and recreational benefits associated with trampoline use, papers in the general pediatric population mostly document an increased incidence of injuries, ranging from minor trauma to spinal cord injuries and even death. The aim of this review is to examine the accumulated published data regarding the use of trampolines, to assess their potential contributions and disadvantages for CF patients, and to define whether trampoline use should be recommended. An extensive search in the published medical literature retrieved approximately 60 articles that primarily dealt with trampolines, out of which only two dealt with CF. The preponderance of these articles are reports pertaining to injuries related to the use of trampolines, with only a few describing the medical, physiologic, and/or psychological benefits of trampolines. Based on the accumulated data, the presumed benefits of trampoline use for CF patients are not proven. Furthermore, the suggested benefits could be acquired using other types of exercise. Weighing the known risks of trampolines against the potential benefits that are not unique to this modality suggests that the use of trampolines for CF should not be recommended. PMID- 15532081 TI - French experience of silicone tracheobronchial stenting in children. AB - Silicone stents were inserted into the trachea or left main-stem bronchus in 14 children aged 2-69 months (median, 7 months). Indications were as follows: tracheomalacia or airway kinking (7 cases), vascular compression (5 cases), and surgically corrected congenital tracheal stenoses (2 cases). The best results were obtained in tracheomalacia. Overall, 6 cases out of 14 (43%) were considered successful, with a stent placement duration of 3-15 months (median, 7 months). Two cases were considered a technical success, although they were clinical failures. Five cases were considered failures primarily due to stent migration. A retrospective analysis of failures suggests that most of these could have been avoided by the use of larger stents. One patient died of stent obstruction. No wall erosion was observed, and the development of granulation tissue was infrequent. Endoscopic removal of the prostheses was uneventful. The biocompatibility of silicone stents appears to be better than what is reported for metal ones, although the stability of the former is less satisfactory. The present study shows the feasibility of silicone stent placement in infants. These stents should be considered as a possible therapeutic option in certain types of childhood airway disorders, although further studies are required. PMID- 15532082 TI - The use of quantile regression in health care research: a case study examining gender differences in the timeliness of thrombolytic therapy. AB - Investigators are frequently interested in determining patient and system characteristics associated with delays in the provision of essential medical treatment. Investigators have typically used either multiple linear regression or Cox proportional hazards models to assess the impact of patient and system characteristics on the timeliness of medical treatment. A drawback to the use of these two methods is that they allow, at best, a partial exploration of how a distribution of delays in treatment or of waiting times changes with patient characteristics. In contrast, quantile regression models allow one to assess how any quantile of a conditional distribution changes with patient characteristics. We illustrate the utility of quantile regression by examining gender differences in the delivery of thrombolysis in patients with an acute myocardial infarction. We demonstrate that richer inferences can be drawn through the use of quantile regression. Females were more likely to experience delays in treatment compared to males. Furthermore, gender had a greater impact upon those patients who had the greatest delays in treatment. Investigators who want to determine how a distribution of delays in treatment or of waiting times changes with patient or system characteristics should consider complementing their analyses with the use of quantile regression. PMID- 15532083 TI - Poly(N-isopropylacrylamide) copolymer films as vehicles for the sustained delivery of proteins to vascular endothelial cells. AB - The aim of this study was to establish the capacity of thermoresponsive poly(N isopropylacrylamide) copolymer films to deliver bioactive concentrations of vascular endothelial growth factor (VEGF165) to human aortic endothelial cells (HAEC) over an extended time period. Films were prepared using a 50:50 (w/w) mixture of non-crosslinkable and crosslinkable copolymers of the following monomer compositions (w/w): 85:15, N-isopropylacrylamide (NiPAAm):N-tert butylacrylamide (NtBAAm); and 85:13:2 NiPAAm:NtBAAm:acrylamidobenzophenone (ABzPh, crosslinking agent), respectively. After crosslinking by UV irradiation, the ability of films to incorporate a fluorescently labeled carrier protein (FITC labeled BSA, 1 mg loaded per film), at 4 degrees C, was first established. Incorporation into the matrix was confirmed by the observation that increasing film thickness from 5 to 10 microm increased release from collapsed films at 37 degrees C (1.76 +/- 0.15 and 10.98 +/- 3.38 microg/mL, respectively, at 24 h postloading) and that this difference was maintained at 5 days postloading (1.81 +/- 0.25 and 13.8 +/- 2.3 microg/mL, respectively). Incorporation was also confirmed by visualization using confocal microscopy. When 10-microm films were loaded with a BSA solution (1 mg/mL) containing VEGF165 (3 microg/mL), sustained release of VEGF165 was observed (10.75 +/- 3.11 ng at 24 h; a total of 31.32 +/- 8.50 ng over 7 days). Furthermore, eluted VEGF165 increased HAEC proliferation by 18.2% over control. The absence of cytotoxic species in medium released from the copolymer films was confirmed by the lack of effect of medium (incubated with copolymer films for 3 days) on HAEC viability. In conclusion this study has shown that NiPAAm:NtBAAm copolymers can be loaded with a therapeutic protein and can deliver bioactive concentrations to human vascular endothelial cells over an extended time period. PMID- 15532084 TI - Towards a fully synthetic substitute of alginate: optimization of a thermal gelation/chemical cross-linking scheme ("tandem" gelation) for the production of beads and liquid-core capsules. AB - Fully synthetic polymers were used for the preparation of hydrogel beads and capsules, in a processing scheme that, originally designed for calcium alginate, was adapted to a "tandem" process, that is the combination a physical gelation with a chemical cross-linking. The polymers feature a Tetronic backbone (tetra armed Pluronics), which exhibits a reverse thermal gelation in water solutions within a physiological range of temperatures and pHs. The polymers bear terminal reactive groups that allow for a mild, but effective chemical cross-linking. Given an appropriate temperature jump, the thermal gelation provides a hardening kinetics similar to that of alginate. With slower kinetics, the chemical cross linking then develops an irreversible and elastic gel structure, and determines its transport properties. In the present article this process has been optimized for the production of monodisperse, high elastic, hydrogel microbeads, and liquid core microcapsules. We also show the feasibility of the use of liquid-core microcapsules in cell encapsulation. In preliminary experiments, CHO cells have been successfully encapsulated preserving their viability during the process and after incubation. The advantages of this process are mainly in the use of synthetic polymers, which provide great flexibility in the molecular design. This, in principle, allows for a precise tailoring of mechanical and transport properties and of bioactivity of the hydrogels, and also for a precise control in material purification. PMID- 15532086 TI - Explained randomness in proportional hazards models. AB - A coefficient of explained randomness, analogous to explained variation but for non-linear models, was presented by Kent. The construct hinges upon the notion of Kullback-Leibler information gain. Kent and O'Quigley developed these ideas, obtaining simple, multiple and partial coefficients for the situation of proportional hazards regression. Their approach was based upon the idea of transforming a general proportional hazards model to a specific one of Weibull form. Xu and O'Quigley developed a more direct approach, more in harmony with the semi-parametric nature of the proportional hazards model thereby simplifying inference and allowing, for instance, the use of time dependent covariates. A potential drawback to the coefficient of Xu and O'Quigley is its interpretation as explained randomness in the covariate given time. An investigator might feel that the interpretation of the Kent and O'Quigley coefficient, as a proportion of explained randomness of time given the covariate, is preferable. One purpose of this note is to indicate that, under an independent censoring assumption, the two population coefficients coincide. Thus the simpler inferential setting for Xu and O'Quigley can also be applied to the coefficient of Kent and O'Quigley. Our second purpose is to point out that a sample-based coefficient in common use in the SAS statistical package can be interpreted as an estimate of explained randomness when there is no censoring. When there is censoring the SAS coefficient would not seem satisfactory in that its population counterpart depends on an independent censoring mechanism. However there is a quick fix and we argue in favour of its use. PMID- 15532087 TI - Pulmonary capillaritis with hemorrhage due to propylthiouracil therapy in a child. AB - We report on a case of pulmonary capillaritis with diffuse alveolar hemorrhage in a child due to propylthiouracil (PTU). PTU treatment is a rare cause of pulmonary capillaritis in adults; we report on the first case in a pediatric patient. The treatment of pulmonary capillaritis often requires corticosteroid therapy, other immunosuppressive medications, or withdrawal of the causative agent. Our patient recovered completely after treatment with a limited course of corticosteroids and removal of PTU. PMID- 15532088 TI - Right diaphragmatic hernia and hydrops: is it always fatal? PMID- 15532089 TI - A comparison of intent-to-treat and per-protocol results in antibiotic non inferiority trials. AB - While the intent-to-treat (ITT) analysis is widely accepted for superiority trials, there remains debate about its role in non-inferiority trials. It is often said that the ITT tends to be anti-conservative in the demonstration of non inferiority. This concern has led to some reliance on per-protocol (PP) analyses that exclude patients on the basis of post-baseline events, despite the inherent bias of such analyses. We compare ITT and PP results from antibiotic trials presented to the public at the FDA's Anti-infective Drug Advisory Committee from 1999 to 2003. While the number of available trials is too small to produce clear conclusions, these data did not support the assumption that the ITT would lead to smaller treatment difference than the PP, in the setting of antibiotic trials. Possible explanations are discussed. PMID- 15532090 TI - Testing the equality of two Poisson means using the rate ratio. AB - In this article, we investigate procedures for comparing two independent Poisson variates that are observed over unequal sampling frames (i.e. time intervals, populations, areas or any combination thereof). We consider two statistics (with and without the logarithmic transformation) for testing the equality of two Poisson rates. Two methods for implementing these statistics are reviewed. They are (1) the sample-based method, and (2) the constrained maximum likelihood estimation (CMLE) method. We conduct an empirical study to evaluate the performance of different statistics and methods. Generally, we find that the CMLE method works satisfactorily only for the statistic without the logarithmic transformation (denoted as W(2)) while sample-based method performs better for the statistic using the logarithmic transformation (denoted as W(3)). It is noteworthy that both statistics perform well for moderate to large Poisson rates (e.g. > or =10). For small Poisson rates (e.g. <10), W(2) can be liberal (e.g. actual type I error rate/nominal level > or =1.2) while W(3) can be conservative (e.g. actual type I error rate/nominal level < or =0.8). The corresponding sample size formulae are provided and valid in the sense that the simulated powers associated with the approximate sample size formulae are generally close to the pre-chosen power level. We illustrate our methodologies with a real example from a breast cancer study. PMID- 15532091 TI - Effects of maternal nicotine exposure on lung surfactant system in rats. AB - Maternal smoking during pregnancy may impair pulmonary function in infants and children, but the exact mechanisms underlying these changes remain to be determined. Timed pregnant Sprague-Dawley rats were injected subcutaneously with nicotine at a dose of 2 mg/kg/day from days 3-21 of gestation. A control group was injected with saline. Nicotine-treated dams had lower body weights than control dams from gestational days 5-21, and the values reached statistical significance on gestational days 17, 20, and 21. Total lung saturated phosphatidylcholine contents tended to be lower in nicotine-exposed rats than in control rats from postnatal day 21, and the values reached statistical significance on postnatal days 35 and 42. Maternal nicotine exposure significantly increased surfactant protein (SP)-A, SP-B, SP-C, and SP-D mRNA expression on postnatal day 7, and decreased SP-A, SP-B, SP-C, and SP-D mRNA expression on postnatal day 14. In conclusion, maternal nicotine exposure during pregnancy reduces lung surfactant lipids and produces variable changes in surfactant protein gene expression during the late postnatal period. As good surface activity of pulmonary surfactant is essential for normal lung function, these results suggest that derangement of the pulmonary surfactant system may be important in the pathogenesis of impaired pulmonary function in children exposed in utero to nicotine. PMID- 15532092 TI - Remote intrapulmonary spread of recurrent respiratory papillomatosis with malignant transformation. AB - Recurrent respiratory papillomatosis is the most common neoplasm of the larynx in childhood. Extension into lung parenchyma occurs in less than 1% of patients and has a low risk of malignant transformation. Treatment options for intrapulmonary spread have shown limited success. We describe a case of recurrent respiratory papillomatosis with extensive parenchymal involvement and adenosquamous carcinoma in a 14-year-old girl. PMID- 15532093 TI - Effect of alpha-lipoic acid supplementation on markers of protein oxidation in post-mitotic tissues of ageing rat. AB - In the present study, we investigated whether DL-alpha-lipoic acid (LA) supplementation could have prooxidant or antioxidant effects on oxidative protein damage parameters such as protein carbonyl (PCO), nitrotyrosine (NT), advanced oxidation protein products (AOPP), and protein thiol (P-SH), as well as oxidative stress parameters such as total thiol (T-SH), non-protein thiol (Np-SH), and lipid hydroperoxide (LHP) in the brain and the skeletal muscle tissue of aged rats. PCO, and NT levels were increased, AOPP and P-SH levels were not changed in the brain tissue of aged rats given LA supplementation. On the other hand, TSH, Np-SH, and LHP levels were decreased in the brain tissue of aged rats given LA supplementation. The levels of the same parameters were not significantly different in the skeletal muscle tissue of aged rats given LA supplementation. The increased levels of protein oxidation markers such as PCO, and NT in the brain tissue of LA-supplemented aged rats compared with non-supplemented aged rats may suggest that oxidative protein damage is increased in LA-supplemented aged rats. We assume that an explanation for our findings regarding LA supplementation on protein oxidation markers in the brain tissue of aged rats may be due to the prooxidant effects of LA. Depending on post-mitotic tissue type and dosage of LA, the prooxidant effects of LA supplementation, should be considered in future studies. PMID- 15532094 TI - Suicidal ideation and bipolar-II depression symptoms. AB - OBJECTIVE: The FDA issued a warning about a possible risk of suicidal behaviour related to 'newer' antidepressants. Suicidal behaviour is common in bipolar-II (BP-II) depression, which is frequent in outpatients, and often mixed (i.e. it has concurrent hypomanic symptoms). The study aim was to find major depressive episode (MDE) and intra-MDE hypomanic symptoms associated with suicidal ideation. METHODS: A total of 374 consecutive BP-II MDE outpatients were interviewed by the structured clinical interview for DSM-IV (SCID), the hypomania interview guide, and the family history screen. RESULTS: Suicidal ideation was present in 52.6%. Suicidal ideation and lower GAF (meaning more severity), more persistent MDE symptoms, more melancholic depressions were significantly associated. Multiple logistic regression of suicidal ideation versus MDE symptoms and intra-MDE hypomanic symptoms found, as significant independent predictors, decreased self esteem, racing/crowded thoughts, psychomotor agitation. DISCUSSION: As expected, suicidal ideation and depression severity were associated. Racing/crowded thoughts and psychomotor agitation were independent predictors of suicidal ideation. While cross-sectional associations cannot show a causal association, some evidence from prospective studies supports the direction of the association. As BP-II depression is common and often mixed in outpatients, clinicians should assess intra-MDE racing/crowded thoughts and psychomotor agitation, as antidepressants alone may worsen these symptoms, and this may induce or increase suicidal behaviour. Mood stabilizing agents may be needed to control these excitement symptoms before using antidepressants. PMID- 15532095 TI - Expression profiling of microdissected matched prostate cancer samples reveals CD166/MEMD and CD24 as new prognostic markers for patient survival. AB - In order to screen for differentially expressed genes that might be useful in diagnosis or therapy of prostate cancer we have used a custom made Affymetrix GeneChip containing 3950 cDNA fragments. Expression profiles were obtained from 42 matched pairs of mRNAs isolated from microdissected malignant and benign prostate tissues. Applying three different bioinformatic approaches to define differential gene expression, we found 277 differentially expressed genes, of which 98 were identified by all three methods. Fourteen per cent of these genes were not found in other expression studies, which were based on bulk tissue. Resultant candidate genes were further validated by quantitative RT-PCR, mRNA in situ hybridization and immunohistochemistry. AGR2 was over-expressed in 89% of prostate carcinomas, but did not have prognostic significance. Immunohistologically detected over-expression of MEMD and CD24 was identified in 86% and 38.5% of prostate carcinomas respectively, and both were predictive of PSA relapse. Combined marker analysis using MEMD and CD24 expression proved to be an independent prognostic factor (RR = 4.7, p = 0.006) in a Cox regression model, and was also superior to conventional markers. This combination of molecular markers thus appears to allow improved prediction of patient prognosis, but should be validated in larger studies. PMID- 15532096 TI - Mortalin is over-expressed by colorectal adenocarcinomas and correlates with poor survival. AB - Using comparative proteomic analysis we have identified over-expression of mortalin in colorectal adenocarcinomas. Mortalin, also known as mitochondrial heat-shock protein 70 (mhsp 70), is involved in cell cycle regulation with important roles in cellular senescence and immortalization pathways. It is known to bind to and inactivate wild-type tumour suppressor protein p53 and influences the Ras-Raf-MAPK pathway. By immunostaining a colorectal cancer tissue microarray linked to a patient database, we further found that mortalin over-expression correlates with poor patient survival and, in multivariate analysis, is independent of standard prognostic variables (p = 0.04). Our findings demonstrate that mortalin over-expression may predict outcome in colorectal cancer and suggest that this protein is involved in colorectal neoplasia. Our experimental approach emphasises the analytical power of combining proteomics with tissue microarray analysis in the context of a well-defined tumour database. PMID- 15532097 TI - Uncertainties associated with incidence estimates of HIV/AIDS diagnoses adjusted for reporting delay and risk redistribution. AB - Reporting delay occurs frequently in the case surveillance of a disease such as HIV/AIDS. To evaluate the current trend of incidence, it is important to consider incidence estimates adjusted for reporting delays and the uncertainty associated with this adjustment. For a surveillance system in which cases are reported monthly, there is no straightforward method for constructing the confidence interval estimators for annual incidence or incidence for a period longer than a month. This is because the monthly incidence estimators are not independent and the correlations among them are not available. Furthermore, to estimate the incidence for a specific risk, or exposure, group (e.g. men who have sex with men), we also have to consider the uncertainty associated with the counts from cases reported without an identified risk. Cases with no reported risk are assigned proportionally to each risk group on the basis of experience with cases reported initially with no reported risk but reported later with an identified risk. In this article, we introduce a method for combining the uncertainties associated with both reporting delay and risk redistribution. An estimator for the covariance between two incidence estimators is also provided so that we can make pairwise comparisons and discover any significant changes in incidence over time. Results are applied to evaluating the current trends in AIDS incidence among men who have sex with men in the United States. PMID- 15532098 TI - Optimizing inhaled corticosteroid therapy in children with chronic asthma. AB - Asthma is the most common chronic illness among children, and inhaled corticosteroids (ICS) are the most effective long-term therapy available for suppressing airway inflammation in persistent asthma. While the primary aim of ICS therapy is good efficacy with minimal side effects, early diagnosis and treatment of asthma can also improve asthma control and normalize lung function, and may prevent irreversible airway injury. Poor patient compliance is a major barrier to treatment. Simplified dosing regimens (e.g., once-daily administration), good inhaler technique, and education of the patient/caregiver should improve patient compliance. Concerns over ICS therapy are often based on the potential for systemic effects associated with oral corticosteroids (e.g., effects on bone mineral density, or growth suppression in children). Since adverse events are associated with high doses of ICS, the dose in all patients should be titrated to the minimum effective dose required to maintain control. Optimal distribution of an ICS in the lungs rather than the systemic compartment is affected by several factors, including the drug's pharmacokinetic profile, inhaler type, inhaler technique, and drug particle size. For young patients unable to use a dry-powder inhaler or pressurized metered-dose inhaler, a nebulizer facilitates drug delivery through passive inhalation; ICS therapy in the form of budesonide inhalation suspension can be given to children with persistent asthma from 12 months of age. In conclusion, selecting a drug with good efficacy and minimal side effects, such as budesonide, together with an easy to-use delivery system and ongoing patient/caregiver education, is important in optimizing ICS therapy for children with persistent asthma. PMID- 15532099 TI - Effective non-viral leader for cap-independent translation in a eukaryotic cell free system. AB - The 61 nt 5'-untranslated region (5'-UTR) of mRNA encoding for a light-emitting protein of hydroid polyp Obelia longissima, obelin, is shown to provide a high level of cap-independent translation of heterologous mRNAs in cell-free translation systems based on wheat germ extracts. The inhibition of translation typically observed when excess mRNA is present or produced in a eukaryotic system (the so-called self-inhibition phenomenon) is found abated with mRNA constructs carrying the obelin mRNA leader. The role of the sequestration of a limiting initiation factor, probably eIF4F, in the self-inhibition phenomenon and the possible independence of the obelin mRNA leader from eIF4F are discussed. We propose the obelin mRNA leader be used for effective cap-independent translation in eukaryotic cell-free systems, including combined transcription-translation systems with uncontrolled phage polymerase-catalyzed accumulation of mRNA. PMID- 15532100 TI - Passive smoking does not increase hydrogen peroxide (H2O2) levels in exhaled breath condensate in 9-year-old healthy children. AB - Environmental tobacco smoke, also called passive smoking, was shown to have adverse effects on the health of children. Hydrogen peroxide (H2O2) is proposed as a sensitive marker of oxidative injury and inflammatory processes in the airways, being increased in adult active cigarette smokers. We tested whether passive smoking had an influence on H2O2 exhalation in healthy children. Thirty healthy passive smoking and 24 nonexposed healthy children aged 9 years were included in the study. Exhaled breath condensate (EBC) was obtained by spontaneous tidal volume breathing with EcoScreen (Jaeger, Germany). All subjects underwent flow-volume measurements immediately after EBC collection. Levels of H2O2 were measured fluorimetrically with the homovanillic acid method. Lung function did not differ between the passive smoking and nonexposed children groups. In the passive smoking group, EBC H2O2 concentration (median and range) was 0.32 (0.00-1.20) microM, and did not differ significantly (P >0.05) from that found in the nonexposed group, i.e., 0,22 (0.00-0.68) microM. Exhaled H2O2 did not correlate with spirometric parameters (FEV1, FEV1%FVC, and MEF50%FVC) in either group. We conclude that passive smoking does not increase H2O2 exhalation in healthy children. PMID- 15532101 TI - Statistical modelling of the differences between successive R-R intervals. AB - Understanding the behaviour of R-R interval data and its successive differences is critical to the dynamics of cardiac control. Several time domain measures that quantify R-R interval variability have important clinical significance in terms of risk stratification and evaluating the effectiveness of treatment procedures. The present approach at examining the distributions of successive beat-to-beat differences of R-R interval data from different populations and under different conditions (baseline and reaction times) provides a valuable insight into their previously unexplored distributional properties. In particular, our analysis reveals that the successive differences have non-normal statistical distributions (a contradiction to the commonly used assumption of normality), and the absolute successive R-R interval differences approximately follows a Weibull distribution. As an illustration of the utility of this approach, we explore the statistical properties of the time domain measure: root mean square successive difference, study the association between the Weibull scale parameter estimate and respiratory sinus arrhythmia, and propose improvements in artifact detection algorithms. PMID- 15532102 TI - Effect of retinoic acid on oxygen-induced lung injury in the newborn rat. AB - Oxygen-induced lung injury is believed to lead to the development of bronchopulmonary dysplasia (BPD). To determine whether retinoic acid (RA) treatment prevents the development of BPD by minimizing lung injury, we investigated the effect of RA on the histopathologic characteristics of oxygen induced lung injury in a newborn rat model. Eighteen rat pups were divided into three groups: room air-exposed control group (n=5), oxygen-exposed placebo group (n=7), and RA-treated oxygen-exposed group (n=6). Measurement of alveolar area, quantitation of secondary crest formation, microvessel count, evaluation of alveolar septal fibrosis, and smooth muscle actin (SMA) immunostaining were performed to assess oxygen-induced changes in lung morphology. Treatment of oxygen-exposed animals with RA resulted in a significant increase in mean alveolar area; however, it had no effect on the number of secondary crests and microvessel count. The degree of fibrosis and SMA expression showed a significant decrease in RA-treated animals. We conclude that RA treatment improves alveolar structure and decreases fibrosis in the newborn rat with oxygen-induced lung injury. Extrapolating these findings to humans, we speculate that similar treatment with RA may reduce lung injury in preterm infants at risk for BPD. PMID- 15532103 TI - A biomimetic model of tandem radical damage involving sulfur-containing proteins and unsaturated lipids. PMID- 15532105 TI - [Combined surgery in synchronous cancer of the sigmoid colon and urinary bladder]. PMID- 15532104 TI - Chemical approaches to controlling intracellular protein degradation. PMID- 15532106 TI - [Endovascular grafting of a giant false aneurysm of superficial femoral artery]. PMID- 15532107 TI - [Introduction of pathogenic principles of surgical treatment of ulcer disease in Russian surgery]. PMID- 15532108 TI - Immunosurveillance of alveolar echinococcosis by specific humoral and cellular immune tests: long-term analysis of the Swiss chemotherapy trial (1976-2001). AB - BACKGROUND/AIMS: Long-term chemotherapy with benzimidazoles is beneficial in non resectable alveolar echinococcosis (AE). Criteria to track early therapeutic efficacy are lacking and the clinical impact of immunosurveillance is unsettled. We aimed to analyze this issue particularly for assessing the putative parasitocidal efficacy of chemotherapy. METHODS: The present study is part of our prospective Swiss trial outlined previously and comprises 57 patients with a median follow-up of 18.5 (3-30) years and with repeated tests of humoral and cell mediated immunity. The series was subdivided into group A (n=23; curative surgery) and group B (n=34: non-resectable AE). RESULTS: Long-term survival was 87% (group A) and 76% (group B). The profiles of specific antibodies against EmII/3-10 antigen normalized within 3 years in most group A-patients, but remained above the cut-off value in 40% of group B-patients. This lack of normalization was associated with lower bioavailability of mebendazole. AE recurrence after 'radical' surgery (up to 13 years) was associated with high anti EmII/3-10 concentrations in 7 of 8 cases. Following abrogation of longterm chemotherapy in group B, no AE-recurrence occurred in 9/18 patients, suggestive of parasitocidal efficacy and documented by a normal EmII/3-10 profile. CONCLUSIONS: The EmII/3-10 profile is of value in monitoring AE after surgery and/or chemotherapy. PMID- 15532109 TI - Sutureless plug repair of obturator hernia using a thigh approach. PMID- 15532110 TI - Designing a basic surgical timetable. PMID- 15532112 TI - [Surgery of the thyroid gland. I]. PMID- 15532113 TI - The 'true' acetabular index in children with cerebral palsy. AB - INTRODUCTION: The acetabular index is a measurement used to determine whether or not an acetabular osteotomy may be required at the time of hip reconstruction in children. METHODS: A retrospective analysis of pre- and postoperative values of the acetabular index was carried out on two groups of children. One group of children had an acetabuloplasty and the other did not during hip reconstruction, the decision being made at the time of open reduction, based on lack of hip congruity. RESULTS: When the pre-operative acetabular index measurements were reviewed, the mean difference between the two groups was only 7 degrees. Range of measurements was similar between the two groups. CONCLUSIONS: The acetabular index is difficult to measure in some children with cerebral palsy. The bony landmarks are difficult to standardise. This may explain the similar pre operative values between the two groups of children in this study. We, therefore, recommend that before hip reconstruction in children with cerebral palsy, the measurement of the acetabular index should be abandoned. Decisions as to whether to undertake an acetabuloplasty or not should be made at the time of open reduction. PMID- 15532115 TI - Endovascular repair (EVAR)of abdominal aortic aneurysms: the case against. PMID- 15532116 TI - Evaluation of knot tying at depth between general surgeons and urologists. PMID- 15532117 TI - A technique for reduction and casting of closed tibial fractures. PMID- 15532118 TI - The 'arthroscopy toast rack'--it's all in the presentation. PMID- 15532119 TI - Scrotal support in anorectal surgery--the Merthyr mask. PMID- 15532120 TI - Removal of ruptured cysts. PMID- 15532121 TI - The management evaluation of General Professional Practice Surgery (GPPS). AB - This report was carried out to identify the management structures and processes that could support or disable curriculum innovation in postgraduate medical and dental education (PGMDE). PMID- 15532122 TI - Some problems of permanent artificial pacing. PMID- 15532123 TI - Interactive CD-ROM: making learning easier. PMID- 15532124 TI - National Federation Biographical Nursing Project. PMID- 15532125 TI - NME digest. PMID- 15532126 TI - Spotlight on the effect of statins on stroke. PMID- 15532127 TI - [Learn your whole life to learn]. AB - Lifelong learning is no longer an option for dentists but rather a necessity, an obligation. In this contribution a survey is presented of all persons and institutions involved in the supply for continuing professional development (CPD). Subsequently the role will be discussed of the government, the universities, the professional organizations, the scientific associations and the scientific dental journals. The most decisive factor however is the own responsibility of the individual dental practitioner to keep learning a life long. Future developments will be touched upon shortly. PMID- 15532128 TI - A comparison of the electroencephalogram between institutionalized and community children in Romania. AB - Electroencephalographic (EEG) data were collected from a sample of institutionalized infants and young children in Bucharest, Romania, and were compared with EEG data from age-matched children from the local community who had never been institutionalized and who were living with their families in the Bucharest area. Compared with the never-institutionalized group, the institutionalized group showed a pattern of increased low-frequency (theta) power in posterior scalp regions and decreased high-frequency (alpha and beta) power, particularly at frontal and temporal electrode sites. This finding is consistent with EEG studies of children facing environmental adversity and children with learning disorders. The institutionalized group also showed less marked hemispheric EEG asymmetries than the never-institutionalized group, particularly in the temporal region. The results are discussed in the context of two models: that the pattern of EEG in the institutionalized children reflects a maturational lag in nervous system development, or that it reflects tonic cortical hypoactivation. PMID- 15532129 TI - Polio eradication in India: some observations. AB - In 1988, the World Health Assembly passed resolution WHA 41.28, which committed the World Health Organization (WHO) to the global eradication of poliomyelitis by the year 2000. In spite of the combined efforts by UNICEF, National Polio Surveillance Project (NPSP), Indian Academy of Pediatrics (IAP) and Rotary International, Polio Free India is still a distant dream. Though oral polio vaccine has succeeded in polio eradication from many countries but there is high incidence of vaccine failure in India. Oral polio vaccine (OPV) has failed to provide full protection to many children who have developed paralytic polio even after taking 10 or more doses of OPV. In some children, OPV has caused paralysis vaccine associated paralytic polio (VAPP). Number of children developing polio due to vaccine is high and on increase. Reasons for this could be that even immunocompromised children are being administered OPVbecause IPV is not available. Vaccine failure has exaggerated the problem of VAPP. No efforts have been made to find the causes for high incidence of vaccine failure and VAPP. PMID- 15532130 TI - Inhibition of lipid peroxidation by botanical extracts of Ocimum sanctum: in vivo and in vitro studies. AB - Ocimum sanctum, the Indian holy basil, has significant ability to scavenge highly reactive free radicals. Shade dried leaf powder of the plant was extracted with water and alcohol, and then fractionated with different solvents. Both extracts and their fractions have in vitro anti-lipidperoxidative activity at very low concentrations. In vivo, hypercholesterolemia-induced erythrocyte lipid peroxidation activity was inhibited by aqueous extracts of Ocimum in a dose dependent manner in male albino rabbits. Aqueous extract feeding also provided significant liver and aortic tissue protection from hypercholesterolemia-induced peroxidative damage. PMID- 15532131 TI - [2/5 Behavior therapies]. PMID- 15532132 TI - The Websites of Norwegian hospitals: do they meet national guidelines and patient's expectations? AB - In 2001, the Norwegian National Health Administration introduced a system that allows patients to choose the public hospital at which they are to be treated. This increased the importance of hospital Websites, as they are used by patients to select a service. In 2002 we rated the Websites of general hospitals in Norway in two ways. Rating 1 was based on a Norwegian set of quality criteria for Websites; 16 items were evaluated to give a maximum score of 32. For rating 2, we used the top five items ranked by cancer patients or their relatives in a previous study, which were weighted and gave a maximum score of 27. Fifty-four (82%) of the 66 public general hospitals in Norway had a Website on the Internet. Information about departments and treatments offered was sparse. Information on prices was almost completely absent. About a third of the Websites were searchable. Quality, as scored by summing ratings 1 and 2, was related to the type of hospital. The mean scores were: university hospitals, 38.8; central hospitals, 30.3; local hospitals, 29.2; and other hospitals, 21.1. On the whole, the hospitals' Websites were not impressive, in view of the competition between hospitals for patients. PMID- 15532133 TI - Medicare and communities of color. PMID- 15532134 TI - Ethinylestradiol + norelgestromin: new preparation. Transdermal contraception: no tangible progress. AB - (1) The reference hormone-based contraceptive for women is an oral contraceptive combining ethinylestradiol (about 30 micrograms) and a well-known progestin such as levonorgestrel or norethisterone. (2) A transdermal contraceptive patch delivering 20 micrograms of ethinylestradiol and 150 micrograms of norelgestromin over 24 hours, and designed to be left in place for a whole week, three weeks a month, has recently been marketed in France. (3) Norelgestromin is the active metabolite of norgestimate, which is already available in combined contraceptives but is less well evaluated than some other progestins. Norelgestromin is metabolised by the liver, notably into norgestrel. (4) The clinical evaluation dossier of the new transdermal contraceptive contains data from two comparative unblinded trials, one versus a triphasic combination of oral ethinylestradiol + levonorgestrel, and the other versus oral ethinyl estradiol (20 micrograms) + desogestrel. A third, non comparative trial offers weaker evidence. These three trials included about 3300 women in total, and lasted between 6 and 13 cycles. The patch was about as effective as the comparator contraceptives. (5) In the three main clinical trials, 4.7% of patches had to be replaced because they became unstuck, either completely (1.8%) or partially (2.9%). (6) More women dropped out of the groups using patches (19.9% of the patch group compared with 14.5% of the group taking oral contraceptives in one trial, 29.6% versus 24.3% in the other trial). Women using the patch were more likely than other women to stop their treatment because of adverse events (about 12% versus 5%). (7) Breast discomfort, breast tenderness or pain were reported by 22% of women using the patches and by 9% and 6% of women in the two comparator groups. Women using the patches had slightly longer menstrual periods (5.6 days versus 4.7 days). Reactions at the patch site were reported by 17% of women. (8) There is no evidence that the patch is any less likely than reference oral contraceptives to cause thromboembolism. The true thromboembolic risk associated with the new patches is unknown. (9) Used patches still contain large amounts of active substances, and must be placed in sachets (provided in the packet) and taken to a pharmacy for disposal. (10) In practice, the reference combined contraceptive for women is still oral ethinylestradiol (about 30 micrograms) plus a well-known progestin such as levonorgestrel or norethisterone. Ethinylestradiol + norelgestromin patches offer women no real benefits: they are probably less convenient and may be less safe. PMID- 15532135 TI - Alteplase: new indication. Inadequately assessed in ischaemic stroke. AB - (1) Alteplase is the first thrombolytic drug to be approved in France for the treatment of ischaemic stroke within three hours of symptom onset. (2) The clinical evaluation dossier contains nine placebo-controlled trials, of which six were relatively large. In the two NINDS trials (624 patients in total), treatment was started within the first three hours and it showed no survival benefit. Near complete functional recovery was more frequent in the alteplase group than in the placebo group. In the two ECASS trials (620 and 800 patients) and the ATLANTIS trials (142 patients and 613 patients), treatment was started within the first six hours and it showed no significant benefit in terms of survival or functional recovery. (3) There are two meta-analyses of these trials. They confirm the lack of a survival benefit with alteplase. Using a combined endpoint, one meta analysis showed that treating 1000 patients with alteplase prevented death or major disability (dependency) in 55 patients. The other meta-analysis underlined the importance of a short interval between the onset of symptoms and the beginning of treatment. (4) Intracranial haemorrhage is the most important adverse effect. One meta-analysis showed that alteplase caused 62 additional symptomatic intracranial haemorrhages (including 25 deaths) per 1000 treated patients. (5) Various retrospective subgroup analyses have tentatively identified subgroups of patients at a particularly high risk of adverse effects, but subgroup analyses provide only weak evidence. The patients most likely to benefit from alteplase, started within three hours of symptom onset, remain to be defined. (6) The current health infrastructure in France would allow only a small number of stroke patients to be treated with alteplase under the kind of conditions prevailing in clinical trials (imaging to confirm ischaemic stroke, and treatment very soon after the onset of symptoms). (7) In practice, there is a narrow margin between the wanted and unwanted effects of alteplase. This treatment should be used only by specialised teams and for strictly selected patients. Research must continue, particularly to identify those patients most likely to benefit from alteplase, and those most likely to be harmed in whom thrombolysis is contraindicated. PMID- 15532136 TI - Rosuvastatin: new preparation. Opt for statins with evidence of efficacy on clinical outcome. AB - (1) Simvastatin and pravastatin are the two reference statins for type IIA and type IIB hypercholesterolaemia because they have the best-documented protective effect against cardiovascular events. Simvastatin and pravastatin are also the reference statins for familial heterozygous hypercholesterolaemia, though there is no evidence that they prevent cardiovascular events in this group. Statins are not very effective in familial homozygous hypercholesterolaemia. (2) Rosuvastatin is the sixth statin to arrive on the French market. The fifth, cerivastatin, was withdrawn from the market in 2001 because of serious adverse effects. (3) Rosuvastatin has not been assessed in terms of morbidity or mortality. The results of comparative trials in type IIA and type IIB hypercholesterolaemia suggest that rosuvastatin is slightly more active than simvastatin, pravastatin and atorvastatin on some lipid parameters after a few weeks of treatment. (4) Rosuvastatin has not been compared with simvastatin or pravastatin in familial heterozygous hypercholesterolaemia. One trial showed it to be slightly more effective than atorvastatin on cholesterol levels. According to one trial, rosuvastatin does not appear to be more effective than atorvastatin in homozygous forms. (5) In clinical trials the adverse effects of rosuvastatin were similar to those of other statins, with the exception of renal adverse effects. We don't know whether rosuvastatin is more or less likely than other statins to cause rhabdomyolysis. (6) Clinical trials reported some cases of proteinuria and renal failure suggesting there is a need for more thorough assessment in long-term trials. (7) In practice, statins with the best-documented benefits (simvastatin and pravastatin) should be used first for cardiovascular prevention in patients with hypercholesterolaemia. PMID- 15532137 TI - Arsenic trioxide: new preparation. Acute promyelocytic leukaemia: encouraging results but persistent doubts. AB - (1) Acute promyelocytic leukaemia is a rare disease. There is a high remission rate after combination treatment with tretinoin and anthracycline, but there is no established treatment for refractory or relapsed disease. Further treatment with tretinoin, combined with intensive cytotoxic chemotherapy, seems to give the best results in patients who qualify for this treatment, but assessment is limited. (2) Arsenic trioxide has now been approved for induction of remission and consolidation in patients with refractory or relapsed acute promyelocytic leukaemia. (3) The clinical evaluation dossier that supported the application contains data from two non comparative trials including 12 and 40 patients. A complete haematological response was obtained in 45 (87%) of the 52 patients, and the survival rate among patients in first relapse was 77% after a median follow up of two years. These results are similar to those previously obtained with tretinoin plus intensive cytotoxic chemotherapy. (4) All the patients treated with arsenic trioxide experienced adverse events. This was to be expected given the acute and chronic toxicity of arsenicals. Most events included fatigue, gastrointestinal disturbances, peripheral neuropathies, prolongation of the QT interval; and biochemical disturbances (hypokalaemia, hyperglycaemia, elevated transaminase activity). (5) Like tretinoin, arsenic trioxide can provoke a potentially severe leukocyte activation syndrome. (6) In practice, these encouraging data justify further assessment of arsenic trioxide. This drug is already an option for patients with refractory or relapsed disease who cannot receive tretinoin plus intensive chemotherapy. PMID- 15532138 TI - Topical diclofenac: new preparation. Moderate efficacy in actinic keratosis. AB - (1) Many treatments are available for actinic keratosis. The most widely used is cryotherapy. Topical application of 5% fluorouracil cream is a second-line option. (2) Marketing authorisation has been granted for a topical gel containing 3% diclofenac, a nonsteroidal antiinflammatory drug. The excipients include 2.5% hyaluronic acid. (3) Animal pharmacology studies and in vitro tests show that hyaluronic acid delays the transcutaneous uptake of diclofenac, leading to higher concentrations in the epidermis. (4) No comparative trials with fluorouracil or other drugs have been published. Five trials comparing 3% diclofenac topical gel with its excipient are available. They show that it takes at least two or three months of treatment for the lesions to disappear in one-third to one-half of patients. Indirect comparison suggests that diclofenac is less effective than fluorouracil in terms of lesion disappearance one month after the end of treatment. The subsequent risk of relapse is unknown. (5) Local adverse effects are numerous and frequent, and include contact dermatitis, skin rash, dry skin, desquamation, pruritus, local pain, and paresthesia. Adverse effects can be due to either diclofenac or to the excipient, and seem to be less intense than with topical fluorouracil. (6) Trials have shown that topical diclofenac is effective at a dose of 0.5 g of gel applied twice a day, but this precise dose is unlikely to be used as no measuring device is included in the packaging. (7) In practice, diclofenac topical gel is less effective than fluorouracil, and is only a moderately effective option when fluorouracil is poorly tolerated and when physical treatments such as cryotherapy fail. PMID- 15532139 TI - Gastroschisis and pseudoephedrine during pregnancy. AB - (1) Gastroschisis is a severe defect in the closure of the abdominal wall in newborns. The incidence seems to be increasing. (2) Three case-control studies have examined pseudoephedrine exposure among mothers of children born with gastroschisis. The evidence is weak but consistent. In two studies, pseudoephedrine exposure was associated with a significant odds ratio. In the most recent study, the risk of gastroschisis was increased 4.2-fold among women taking the pseudoephedrine-paracetamol combination (95% confidence interval 1.9 9.2). Paracetamol is not known to cause this malformation. (3) Pseudoephedrine is a sympathomimetic vasoconstrictor contained in many over-the-counter oral decongestants sold in France. (4) In practice, it is best to avoid prescribing, dispensing or using preparations containing pseudoephedrine during pregnancy. PMID- 15532141 TI - Preventing medication errors. AB - (1) 319 adverse effects due to errors occurring during the drug use process, as well as errors spotted before any incident, were collated in French hospitals in 2000-2001 and characterised for their type, cause, contributing factors, and consequences. (2) All stages of the drug use process were implicated in errors: prescription, transcription, preparation, dispensing, administration, and drug monitoring. (3) Dosing errors (often overdose) were the most common. (4) The main causes were poor practices and inadequate knowledge, followed by problems of communication, packaging and confusion over drug names. (5) Contributing factors included failure to follow rules and procedures, inadequate communication or training, interruptions and distractions, and drug storage on the wards. (6) The prevention of adverse effects linked to drug errors requires a commitment to record all problems, analyse them in depth, and enforce safe working practices for all health professionals. PMID- 15532140 TI - Antiretroviral treatment. HIV infection in adults: better-defined first-line treatment. AB - (1) There is still no cure for HIV infection. The short-term treatment aims are to drive viral load below the current detection limit and to increase the CD4+ T cell count, in order to reduce morbidity and prolong survival. (2) Early initiation of treatment has both advantages and disadvantages. If the patient is symptomatic or if the CD4+ T cell count is below 200 per mm3, antiretroviral treatment should be started immediately. If the patient is asymptomatic, the CD4+ T cell count is above 350 per mm3, and viral load is below 50 000 copies/ml, antiretroviral treatment can often be deferred. Other situations should be considered case by case. (3) The benefits of treating symptomatic primary infection have not been adequately documented, and current recommendations diverge. (4) The advent of new antiretroviral drugs has increased the choice, but cross-resistance often limits treatment options. The new antiretroviral family to have emerged since 1999 is the fusion inhibitor; the only representative of this class, enfuvirtide, is reserved for patients with multiple treatment failure. (5) There is broad agreement on the principles of first-line antiretroviral treatment. It should combine at least two nucleoside (or nucleotide) inhibitors of HIV reverse transcriptase and one non nucleoside inhibitor, or at least one HIV protease inhibitor. Comparative studies have now identified the most effective combinations in terms of virological efficacy and tolerability. The combination should be chosen according to its established efficacy, adverse effects, risks of interactions, and convenience. There is no reference combination suitable for all patients. Among the combinations containing a non nucleoside inhibitor, those based on efavirenz are the most effective after 48 weeks of follow-up. There is less agreement on the optimal treatment of pregnant women. (6) Among the HIV protease inhibitor-based combinations, those containing nelfinavir or the lopinavir + ritonavir combination must be taken during meals. The lopinavir + ritonavir combination showed better virological efficacy than nelfinavir in a comparative trial. Experience and safety evaluation are in favour of nelfinavir, but recent American guidelines issued in November 2003 recommend the lopinavir + ritonavir combination. (7) There is no major difference in virological efficacy between non nucleoside inhibitors and protease inhibitors as first line therapy. (8) In combination with protease inhibitors or non nucleoside inhibitors, the best-assessed nucleoside inhibitors (effective for several years) are lamivudine + zidovudine and lamivudine + stavudine. For first-line treatment, combinations consisting of only three nucleoside (or nucleotide) inhibitors are less effective than combinations containing HIV protease inhibitors or non nucleoside inhibitors. (9) Efficacy is monitored on the basis of changes in viral load and the CD4+ T cell count, one month after the beginning of treatment, then about every three months. Attention must be paid to adverse effects, which may necessitate the replacement of the causative antiviral drug, treatment of the adverse effect, treatment modification or, in extreme cases, treatment withdrawal. (10) Treatment failures must be carefully investigated: the cause(s) may include poor adherence, drug interactions, and inadequate plasma drug concentrations. Ongoing antiretroviral regimens do not always have to be modified if treatment failure occurs. (11) Resistance tests can help to determine the most effective alternative in case of virological failure due to drug resistance. The choice of back-up treatments is complex, and is limited by cross-resistance. Multiple lines of treatment fail in about 3% to 4% of patients. PMID- 15532142 TI - Inhaled anesthetics and the role of the pharmacist in today's surgical environment. Introduction. PMID- 15532143 TI - Characteristics of anesthetic agents used for induction and maintenance of general anesthesia. AB - PURPOSE: The characteristics of ideal intravenous (i.v.) and inhaled anesthetic agents; the rationale for inducing anesthesia with i.v. anesthetics (particularly propofol); the rationale for inducing and maintaining anesthesia with inhaled anesthetics; and the advantages and disadvantages of specific inhaled anesthetics, namely, nitrous oxide, desflurane, halothane, isoflurane, and sevoflurane, are discussed. SUMMARY: Propofol has many characteristics of the ideal i.v. anesthetic, including a rapid, smooth induction of anesthesia and rapid clearance from the body. The use of an i.v. anesthetic such as propofol avoids the claustrophobia associated with the inhaled route. Inhaled anesthetics are preferred for maintenance of anesthesia because they allow a more precise control of the anesthetic state and do so at low cost. The ideal inhaled anesthetic agent has ample potency and a low solubility in blood and tissues (rapid recovery from anesthesia), resists physical and metabolic degradation, and protects and does not injure vital tissues. It also does not cause seizures, respiratory irritation, or circulatory stimulation or deplete the ozone layer. It has a low acquisition cost. Nitrous oxide potency is too small to produce anesthesia by itself. Halothane is too soluble and poses a risk of severe hepatotoxicity. Desflurane, isoflurane, and sevoflurane are preferred because they have an adequate potency, appropriate solubility, and minimal to no risk for hepatotoxicity. CONCLUSION: The utilization choice among desflurane, isoflurane, and sevoflurane is made by weighing specific advantages and disadvantages of each agent. The primary determining factors for use are anesthetic potency and control, rate of anesthesia induction, clearance from the body, and adverse effects. PMID- 15532144 TI - Considerations in selecting an inhaled anesthetic agent: case studies. AB - PURPOSE: Product and patient-specific factors that may influence the selection of an inhaled anesthetic agent are discussed in four case studies. SUMMARY: The cardiovascular, respiratory, hepatic, and renal effects of various inhaled anesthetic agents vary and may be important in selecting an agent for patients with impairment of these systems. The extent to which anesthetics are metabolized and the risk of hepatotoxicity also vary. Halothane is associated with a relatively high risk for arrhythmias and hepatotoxicity. It also has a high solubility in tissues and is associated with a slow onset and termination of effect and recovery. Pharmacokinetic and pharmacodynamic differences among anesthetics may be important considerations, depending on the surgical setting, anticipated duration of surgery, whether endotracheal intubation is used, and other patient-specific factors such as body weight and age. The duration of surgery is an important consideration for most patients because differences between inhaled anesthetics in offset of effect vary with the duration of the procedure. Desflurane or sevoflurane may be preferred for obese patients because of a faster recovery and greater respiratory safety compared with isoflurane. Sevoflurane is preferred for pediatric patients undergoing induction by inhalation because of its low propensity to cause respiratory irritation. CONCLUSION: There are many product- and patient-specific factors that pharmacists must take into account when selecting an inhaled anesthetic agent. It is important to consider all of these factors when developing the pre- and post anesthesia plan. PMID- 15532145 TI - Pharmacoeconomics of inhaled anesthetic agents: considerations for the pharmacist. AB - PURPOSE: Types of economic analyses used for inhaled anesthetic agents, factors to consider in calculating the cost of inhaled anesthetics, limitations of pharmacoeconomic studies of these agents, and strategies for controlling inhaled anesthetic costs are discussed. SUMMARY: Inhaled anesthetic agents comprise a substantial component of drug budgets. Calculation of the cost of administering an inhaled anesthetic should take into consideration the cost per mL, potency, waste, concentration and duration of gas delivery, fresh gas flow rate, molecular weight, and density. The use of newer inhaled anesthetic agents with low solubility in blood and tissue provides a more rapid recovery from anesthesia than older, more soluble agents, and also provides the same level of control of depth of anesthesia at a lower fresh gas flow rate and possibly a lower cost than older agents at a higher fresh gas flow rate. A more rapid recovery may facilitate fast-track recovery and yield cost savings if it allows the completion of additional surgical cases or allows a reduction in personnel overtime expenses. Interpretation of pharmacoeconomic studies of inhaled anesthetics requires an appreciation of the limitations in methodology and ability to extrapolate results from one setting to another. Pharmacists' efforts to reduce anesthetic waste and collaborate with anesthesiologists to improve the use of these agents can help contain costs, but improving scheduling and efficiency in the operating room has a greater potential to reduce operating room costs. CONCLUSION: Much can be done to control costs of anesthetic agents without compromising availability of these agents and patient care. PMID- 15532146 TI - Research in the aged. PMID- 15532147 TI - Health assessments. PMID- 15532148 TI - The doctor's bag. PMID- 15532149 TI - Encounters with older patients in Australian general practice. AB - The BEACH program is a continuous national study of general practice activity in Australia. The content of encounters with patients aged 65 years and over was examined to identify the morbidities and management of older people in general practice in Australia. This provides a backdrop against which the theme articles in this issue of Australian Family Physician can be further considered. PMID- 15532150 TI - Prescribing in older people. AB - BACKGROUND: Prescribing medications to older people is difficult due to comorbidity, limited evidence for efficacy, increased risk of adverse drug reactions, polypharmacy, and altered pharmacokinetics. OBJECTIVE: This article describes the principles underlying clinical geriatric pharmacology including approaches to evaluating the evidence for risk and benefit, and adjusting dose for age related pharmacokinetic changes. DISCUSSION: The challenge for the general practitioner is to balance an incomplete evidence base for efficacy in frail, older people against the problems related to adverse drug reactions without denying older people potentially valuable pharmacotherapeutic interventions. PMID- 15532151 TI - Normal cognitive changes in aging. AB - BACKGROUND: Cognitive abilities decline with normal aging. Fluid abilities such as speed and problem solving show declines from early adulthood, and crystallised abilities such as knowledge and expertise increase until old age. OBJECTIVE: This article reviews changes in normal cognitive aging a nd associated risk factors. It aims to provide a framework for approaching the differentiation of normal cognitive aging from dementia through awareness of mental status screening and changes from previous functioning. DISCUSSION: Understanding normal cognitive aging is essential for assessing older patients presenting with memory and other cognitive complaints. Many of the risk factors for cognitive aging are modifiable such as hypertension, diabetes, and levels of physical, mental and social activity. At a population level, primary prevention or reduction of cognitive aging is possible through addressing modifiable risk factors. This in turn may reduce population levels of dementia. PMID- 15532152 TI - Diagnosis and management of dementia in general practice. AB - BACKGROUND: The assessment and management of dementia has changed in the past 5 years with the introduction of new drugs to combat dementia. In addition, services available for patients with dementia and the means of accessing these services is constantly changing. OBJECTIVE: This article discusses the assessment and management of dementia using two case studies. The case studies raise issues about screening and accessing the new drugs for a patient with early dementia, care planning, and the involvement of the carer for a patient with more advanced disease. DISCUSSION: As the population ages, dementia is an increasingly important part of general practice. It is important general practitioners are aware of early detection issues, and understand basic screening as well as the possibility of accessing new treatments and community management for patients with dementia. PMID- 15532153 TI - Health assessment of elderly patients. AB - BACKGROUND: Comprehensive health assessment of the elderly in primary care will become a more important element of general practice as population aging proceeds and evidence of efficacy emerges. OBJECTIVE: This article describes approaches to provision of health assessment in primary care based on the best available evidence. DISCUSSION: Current best practice includes the use of structured health assessment protocols, an integrated multidisciplinary approach, targeting patient groups with intermediate levels of disability and handicap, in-home assessments and carefully structured follow up mechanisms. PMID- 15532154 TI - Malnutrition in older people--screening and management strategies. AB - BACKGROUND: Malnutrition in older people is not only common, but frequently overlooked. It can result in multiple medical complications, hospitalisation and even death. OBJECTIVE: This article focusses on the consequences, pathophysiology, diagnosis and management of this clinical syndrome. DISCUSSION: Nonphysiological causes of malnutrition include social, psychological, medical and medication related factors. With vigilance, early screening and the institution of simple therapeutic measures of correcting nonphysiological causes and following dietary guidelines for older people, the adverse effects of malnutrition may be minimised. PMID- 15532156 TI - Advance care planning and end of life decision making. AB - BACKGROUND: Aging populations with greater rates of cognitive decline demand increased attention to the issues of end of life decision making and advance care planning (ACP). Legislatures have passed statutes that recognise the necessity for both substitute decision making and the declaration in advance of wishes relating to health care. OBJECTIVE: This article discusses ACP and the role of the general practitioner. DISCUSSION: Advance care planning provides patients, relatives and doctors with greater confidence about the future. There is good evidence that patients desire to discuss end of life care, and GPs are in a good position to engage their patients in considering the issues. They have a responsibility to confirm the decision making capacity of patients who write advance health directive documents, to inform and educate their patients about future health care, and to keep the ACP conversation going. PMID- 15532155 TI - Elder abuse. AB - BACKGROUND: Elder abuse is a common and yet often unrecognised problem in our community. With up to 5% of the community dwelling older population being victims of abuse, the general practitioner has a pivotal role in identifying this abuse. OBJECTIVE: This article provides an outline of the definition of elder abuse, describes the types of abuse seen and the reasons for occurrence of abuse. It summarises the role of the GP in the identification and management of abuse and provides guidance on intervention strategies. Case studies are used to illustrate the issues discussed. DISCUSSION: Elder abuse is defined as any pattern of behaviour which causes physical, psychological, financial or social harm to an older person. The role of the GP in identifying abuse is critical. The vast majority of older people visit their GP at least once a year, and the GP often has a long standing relationship with their patient and the patient's family. They are therefore ideally placed to identify elder abuse. PMID- 15532157 TI - Alcohol and other drug use in later life. AB - BACKGROUND: Problem alcohol and other drug use is rarely suspected in the elderly. However, the elderly are more susceptible to problems related to drug use at lower doses because of age related changes, comorbidities and polypharmacy. Like other age groups, the elderly sometime seek the use of alcohol and other drugs, to help them feel better or to avoid negative feelings such as loneliness and depression. Drug use in the elderly is associated with significant morbidity anid can be masked by other medical problems. OBJECTIVE: This article demonstrates a gradual onset alcohol problem in a longstanding elderly patient, in whom changes in drug or alcohol use may not be easily detected. It also highlights some key differences in drug related presentations in the elderly. DISCUSSION: Some health professionals consider intervention for problem alcohol or other drug use in the elderly ineffective. However, because of the high prevalence of polypharmacy and comorbid pathology, intervention is more likely to result in a significant health improvement. General practitioners have a high level of contact with and are trusted by the elderly, giving them an unparalleled opportunity among health professionals to intervene in problem alcohol and other drug use. PMID- 15532158 TI - Managing adult sexual assault. AB - BACKGROUND: It is estimated that only 15-20% of women who have been sexually assaulted report to police and therefore the real incidence of sexual assault in males and females is unknown. Once reported, acutte cases of sexual assault (within 72 hours of the allegation) may undergo a forensic medical examination to document injuries, collect forensic specimens and provide an opinion to be used by the criminal justice system. OBJECTIVE: This article outlines the process of a forensic medical examination as well as providing a management flowchart for medical practitioners who are caring for a victim of sexual assault who does not wish to report to police. DISCUSSION: Dealing with a sexual assault case is easier and more efficient when the treating doctor has a good understanding of the issues involved in adult sexual assault and how to obtain crisis care for the victim. Early management of a victim of sexual assault, regardless of whether they want to report to police, is important for minimising associated risks (sexually transmitted infections, pregnancy, psychological sequelae), and documentary injuries and obtaining forensic specimens. PMID- 15532159 TI - Toxic epidermal caused by lamotrigine. AB - BACKGROUND: The use of lamotrigine is increasing. Many general practitioners prescribe it in the treatment of epilepsy and as a mood stabilising agent. It has also been suggested as the agent of choice in pregnant women with epilepsy. OBJECTIVE: This article describes the case histories of two patients who present with oral lesions and an extensive rash after starting treatment with lamotrigine. DISCUSSION: Toxic epidermal necrolysis (TEN) is a potentially fatal adverse mucocutaneous drug reaction. Simple maculopapular rashes occur commonly with lamotrigrine. Although rare, serious mucocutaneous reactions such as TEN can occur soon after commencement of treatment with lamotrigine, or after a dose increase in patients already established on treatment. The risk can be minimised by commencing at a low dose, avoiding rapid increases in dosage, and avoiding concomitant use of sodium valproate. PMID- 15532160 TI - Poststroke depression. AB - BACKGROUND: Poststroke depression (PSD) is common and often unrecognised. The diagnosis can be difficult due to deficits of stroke such as impaired self reporting and cognition, poor insight and dysphasia. Untreated PSD can interfere with recovery and adversely affect functional and social outcomes. OBJECTIVE: This article outlines the diagnosis, pathophysiology and treatment for PSD. DISCUSSION: The natural history of PSD suggests that most PSD is not immediate but develops over months with peak prevalence between 6 and 24 months, and in some cases persists up to 3 years following stroke. General practitioners and treating specialists need to actively monitor patients for PSD. While antidepressant medication is the mainstay of treatment for PSD, psychotherapeutic interventions are important. Treatment should include patient and family education, reestablishment of sleep pattern, addressirng functional difficulties, increasing community participation, improving diet and regular exercise. PMID- 15532161 TI - Presbyopia case study. Eye series-20. PMID- 15532162 TI - Where there's a will... PMID- 15532163 TI - Australian consumers' expectations for expanded nursing roles in general practice -choice not gatekeeping. AB - While research has focussed on the current and potential role of the practice nurse (PN) in Australia, the acceptability of this role by consumers has not been investigated. In 2002, two independent studies were carried out into consumer perceptions of PNs. A disscussion between the two groups of researchers at the inaugural National Pratice Nurse Conference in 2003, identifiied significant similarities in the findings of the tese studies. This article reports the combined findings as they relate to consumer's desire for access to the health practitioner of their choice, and their concern that PNs may assume a Gatekeeping role. These perceptions may have significant impact on the acceptance of PNs across Australia. PMID- 15532164 TI - Mortal dread--So you're telling me there's no cure? That I'm going to die? PMID- 15532165 TI - Role play and medical education. PMID- 15532166 TI - A guide to using role plays in registrar teaching. PMID- 15532167 TI - How do Australian GPs manage shoulder dysfunction? PMID- 15532168 TI - Experiences of viscosupplementation for knee osteoarthritis. PMID- 15532169 TI - [Skin contact: protecting and caring for hands]. PMID- 15532170 TI - [Nosocomial urinary tract infection during long-term inpatient nursing: omitting the bladder catheter is the best prevention]. PMID- 15532171 TI - [Short-term nursing: a missed opportunity]. PMID- 15532172 TI - [Inpatient nursing care--3: What is in the future?]. PMID- 15532173 TI - [Expert standard for discharge management in nursing: thinking beyond the hospital door]. PMID- 15532174 TI - [Creating patient-oriented processes: only thinking of cost, limits care]. PMID- 15532175 TI - [Organizing a place for nursing care transfer: wounds become healing sites]. PMID- 15532176 TI - [Nursing care transition: expert standard provides the framework]. PMID- 15532177 TI - [Reform in health care--a chance for nursing? "Many are too quiet" (interview by Katrin Balzer)]. PMID- 15532178 TI - [Conservative therapeutic procedures in urinary incontinence: nursing is more than supplying incontinence aids]. PMID- 15532179 TI - [Nursing visit and client participation: defining roles anew]. PMID- 15532180 TI - [Presentation of statistical data in nursing 10--Box plots: ideal for group comparisons]. PMID- 15532181 TI - [Nursing in Europe--Slovakian Republic: separate council sets the path for the future]. PMID- 15532182 TI - [Spiritual dimensions of nursing care--4: Belief in the Trinity]. PMID- 15532183 TI - [From case to case: silent pain]. PMID- 15532184 TI - [Health promotion in the elderly--a new perspective in geriatric nursing?]. PMID- 15532185 TI - [The Swiss HIV Cohort Study: from research to health care]. PMID- 15532187 TI - [Prevention of vertical HIV transmission--a success story]. AB - Thanks to very effective interventions vertical transmission of HIV has been reduced from over 20% ten years ago to less than 2% today in industrialised countries. This progress has been achieved by combined application of different strategies including antiretroviral treatment of pregnant women, elective caesarian section (prior to labour and rupture of membranes) and refraining from breastfeeding. Fortunately, the Swiss Mother & Child HIV Cohort Study (MoCHiV) has been able to support this evolution with several important contributions. Nevertheless the most important challenge in the prevention of vertical HIV transmission remains to be resolved in this decade. This impressive reduction in vertical transmission achieved in the industrialised world and delineated in this article still needs to be carried forward to those countries in the third world where HIV prevalence is much higher and interventions therefore much more needed. PMID- 15532186 TI - [The Swiss HIV Cohort Study--resource for epidemiology and public health]. AB - The epidemiological situation in Switzerland was characterised by a substantial decline in the number of AIDS cases and HIV-related deaths following the introduction of highly active antiretroviral therapy (HAART). The number of new HIV infections, has, however increased in recent years and AIDS cases and related deaths are no longer declining. We discuss three examples of issues that are relevant to epidemiology and public health in order to illustrate the importance of the study in this context: (i) use of SHCS data to inform the interpretation of routine surveillance data based on reports from physicians and laboratories to the Federal Office of Public Health (FOPH); (ii) analyses of access to and utilization of HAART across different groups of study participants, including migrant populations, those with lower socio-economic status and patients infected through intravenous drug use; and (iii) the monitoring of trends in self-reported sexual behaviour, including condom use with stable and casual partners. The examples demonstrate that the study is an important resource for epidemiological analysis and public health surveillance and action in Switzerland. PMID- 15532188 TI - [SHCS and the laboratory diagnosis of HIV infection--from the development of the HIV Western blot to virus quantification and clinically relevant individual virus characterization]. AB - The first reliable diagnostic tools for the diagnosis of HIV infection, Western blot and ELISA, were created 20 years ago in the U.S., using initially sera from AIDS patients diagnosed clinically in Switzerland. In Swiss laboratories the diagnosis of HIV infection today is established by using 4th generation screening tests which detect both antibodies and p24 antigen while in the doctor's surgery a rapid antibody assay is used. Confirmation in authorized confirmatory labs relies on a set of different minimal combinations of positive test results derived from both the first and second blood specimen. In patients with a confirmed diagnosis of HIV infection, two further principal questions arise concerning, on the one hand, the virus load and, on the other hand, some clinically relevant properties of the infecting virus (type or subtype, resistance against antiretroviral drugs). The paper starts with a personal flashback to the first days of HIV diagnostics and describes the sensible use of tests available today for answering the above-mentioned principal questions. Alternative methods which can be used when standard tests fail or appear unreliable are mentioned also. PMID- 15532189 TI - [Primary HIV infection]. AB - Acute HIV infection presents as a mononucleosis-like disease with fever, lymphadenopathy, myalgias, rash and headache. Other symptoms are occurring at lower frequency. Since acute HIV infection is associated with high viral loads it is important to diagnose patients with this entity, in order to prevent transmission. It is still controversial whether very early therapy during acute seroconversion is beneficial on the long-run. Small studies may indicate that early therapy which is followed by a stop of antiretroviral therapy after one to two years may be followed by a better control of viremia. PMID- 15532190 TI - [The genetics HIV cohort of the Swiss HIV Cohort Study--facing the new challenges in medicine]. AB - Host genetic factors determine the individual natural course of HIV infection and influence the response to therapy and the occurrence of adverse events to treatment. Variants of multiple genes are associated with faster but also slower development of severe immunodeficiency. However, only very rarely the variant of one single gene explains a specific clinical phenotype. But multiple genetic marker form a complex trait, which is difficult to analyse biostatistically. Research in this rapidly evolving field asks for structures in which hypotheses can be generated and evaluated and which combine basic and diagnostic and therapeutic research. The large amount of prospective information on HIV disease natural history and treatment response of the Swiss HIV Cohort Study will make of the Genetics project an excellent test-setting for some of the immediate difficulties in this research field: validation of new markers and modelling of complex traits. PMID- 15532191 TI - [Simplified treatments for HIV infection]. AB - The treatment of HIV infection is reputedly complicated. Indeed, in 1996 and 1997, the most widely used regimen implied five separate pill ingestions every day: A heavy burden even without side effects. Since then, however, treatment has been greatly simplified. The most widely used regimen in Switzerland is the combination of zidovudine, lamivudine and efavirenz, with 1 pill in the morning and 2 pills in the evening. The Swiss HIV Cohort has shown that simplifying therapy increases the success of HIV treatment. Once daily administration will soon be part of standard care, as clinical trials have shown promising results. In the future, many patients may well treat their HIV infection intermittently. Scheduled treatment interruptions are under intensive investigation in Switzerland and elsewhere. Large scale studies with long follow-up are necessary and the first results can not be expected before the end of 2005. PMID- 15532192 TI - [Opportunistic diseases in HIV infection]. AB - Opportunistic diseases have been and still are a major cause of HIV associated morbidity and mortality. Patients with undiagnosed HIV infection often present with symptoms of opportunistic infections. Therefore, all clinicians should know their clinical presentations. Patients with severe immunodeficiency, manifested by low CD4-lymphocyte count, should receive primary prophylaxis against Pneumocystis jiroveci, T. gondii und M. avium. After successful initial treatment of opportunistic infections maintenance therapy is indicated. The introduction of potent antiretroviral combination therapy has led to a decline of incidence and letality of most opportunistic diseases and allows discontinuation of established primary prophylaxis and maintenance therapy if a durable rise of CD4-lymphocyte count can be achieved. The improvement of immunocompetence due to antiretroviral therapy is also associated with new clinical manifestations of opportunistic infections, the immune reconstitution inflammatory syndromes. The framework of the multicenter Swiss HIV Cohort Study with its detailed documentation of opportunistic diseases has allowed several important studies and analyses that have had a worldwide influence on the management of these diseases. PMID- 15532193 TI - [Adherence and resistance]. AB - With the introduction of highly efficacious HAART therapy for HIV/AIDS successful long-term therapy became available. Nevertheless, it also revealed the fragile nature of antiretroviral therapy: resistant virus variants emerged, sometimes already after only short treatment episodes. And they soon began decreasing the newly gained armamentarium of specific inhibitors for HIV. A primary cause for failure could be found in a poor adherence to therapy or in the insufficient metabolic availability of a drug. It is primarily this fact that allows the HI virus, although delayed, to complete its life cycle and to adapt to those incompletely controlling drug levels. And finally the conditioned virus will fail to respond to the respective drug. For the clinical therapy concept it can be fatal if a resistance development remains undetected. At this point the recently established diagnostic tool of resistance-determination is gaining importance: it allows detection and characterization of viral resistances early after emergence, helping to adapt and optimize HAART therapy. Two distinct methodologies have been developed: Genotyping and viral Phenotyping. Genotyping is technically less demanding and can principally be performed within one week. It is based on the identification of (known) resistance-associated mutations and on their combination onto one "virtual viral genome". This can be compared with known resistance-patterns and allows then the extrapolation of a viral resistance. The more demanding phenotyping method represents a direct determination, utilizing the concept of an "in vitro therapy": every drug to be assessed will be directly tested against virus information from the respective patient. As a consequence phenotyping has, particularly in its special format of a "replicative phenotyping", special utility for mixed virus populations or emerging minorities in the same patient, for viruses with new (unknown) mutations, and for viruses with reduced replicative capacity. Either method for the determination of HIV resistance reflects still a relatively young diagnostic tool. As a consequence current studies, underway in Switzerland and supported by the SHCS, aim at the further optimization of the tests and at an evaluation of their long term therapeutic benefit. PMID- 15532194 TI - [The Swiss HIV Cohort Study--multifaceted network supporting patient care as well as large international research projects]. AB - The Swiss HIV Cohort Study (SHCS) integrates networks at several different levels. Initiated as a national collaboration of five University and a Cantonal hospital the SHCS currently also integrates numerous regional hospitals and specialized physicians in private practice. The standardized anonymous data collection allowed a variety of scientific projects--even beyond strictly medical domains such as psychosocial and legal aspects. Although the SHCS is one of the largest HIV cohorts worldwide some important medical questions cannot readily be answered without collaborating in international networks. These networks on different levels facilitate an efficient information exchange aimed at an optimal and standardized care of patients with HIV and AIDS in Switzerland. PMID- 15532195 TI - Is there a sleep debt? PMID- 15532196 TI - Sleep debt and scientific evidence. PMID- 15532197 TI - Sleep apnea syndrome, endothelial dysfunction, and cardiovascular morbidity. PMID- 15532198 TI - Sleepiness combined with low alcohol intake in women drivers: greater impairment but better perception than men? AB - OBJECTIVES: We have previously shown that low blood alcohol concentrations (BAC) (at approximately half the legal driving limit in both the United Kingdom and in most states in the United States) exacerbate moderate sleepiness (sleep during the night restricted to 5 hours) and markedly impair driving ability in young men. There are distinct physiologic sex differences in the absorption, metabolism, and central nervous system effects of alcohol; therefore, we replicated this earlier study, this time using women and using similar BAC to provide a comparison. DESIGN: 2 x 2 repeated-measures counterbalanced. SETTING: 2 hour drive from 2:00 pm in an instrumented car on a simulated highway. INTERVENTIONS: Alcohol versus control and normal sleep versus sleep restricted to 5 hours. MEASUREMENTS AND RESULTS: Driving impairment (lane drifting), subjective sleepiness, and electroencephalographic measures of sleepiness. Sleep restriction significantly worsened driving performance and subjective sleepiness as it had in men. Surprisingly, unlike men, women showed no apparent adverse effects of alcohol alone on these indexes; they seemingly compensated for the effects of alcohol. However, alcohol's effects were profound when alcohol was combined with sleep restriction; nevertheless, women, unlike men, were aware of this enhanced sleepiness. After alcohol ingestion, the electroencephalogram showed increased beta activity, an effect not seen in men, indicating a differential pharmacokinetic effect of alcohol on the central nervous system, compensatory effort, or both. Debriefing questionnaires indicated that women were aware of the varying risks of driving under these different conditions. CONCLUSIONS: Legally "safe" BAC markedly worsen sleepiness-impaired driving in women. However, they seem to be aware of their impaired driving and are able to judge the degree of risk entailed. Such an attitude may contribute to the lower incidence of sleep- or alcohol-related crashes in women compared with men. PMID- 15532199 TI - Low levels of alcohol impair driving simulator performance and reduce perception of crash risk in partially sleep deprived subjects. AB - STUDY OBJECTIVES: Partial sleep deprivation and alcohol consumption are a common combination, particularly among young drivers. We hypothesized that while low blood alcohol concentration (<0.05 g/dL) may not significantly increase crash risk, the combination of partial sleep deprivation and low blood alcohol concentration would cause significant performance impairment. DESIGN: Experimental SETTING: Sleep Disorders Unit Laboratory PATIENTS OR PARTICIPANTS: 20 healthy volunteers (mean age 22.8 years; 9 men). INTERVENTIONS: Subjects underwent driving simulator testing at 1 am on 2 nights a week apart. On the night preceding simulator testing, subjects were partially sleep deprived (5 hours in bed). Alcohol consumption (2-3 standard alcohol drinks over 2 hours) was randomized to 1 of the 2 test nights, and blood alcohol concentrations were estimated using a calibrated Breathalyzer. During the driving task subjects were monitored continuously with electroencephalography for sleep episodes and were prompted every 4.5 minutes for answers to 2 perception scales-performance and crash risk. MEASUREMENTS AND RESULTS: Mean blood alcohol concentration on the alcohol night was 0.035 +/- 0.015 g/dL. Compared with conditions during partial sleep deprivation alone, subjects had more microsleeps, impaired driving simulator performance, and poorer ability to predict crash risk in the combined partial sleep deprivation and alcohol condition. Women predicted crash risk more accurately than did men in the partial sleep deprivation condition, but neither men nor women predicted the risk accurately in the sleep deprivation plus alcohol condition. CONCLUSIONS: Alcohol at legal blood alcohol concentrations appears to increase sleepiness and impair performance and the detection of crash risk following partial sleep deprivation. When partially sleep deprived, women appear to be either more perceptive of increased crash risk or more willing to admit to their driving limitations than are men. Alcohol eliminated this behavioral difference. PMID- 15532200 TI - Effects of sleep deprivation on spontaneous arousals in humans. AB - STUDY OBJECTIVES: The hierarchical definition of arousals from sleep includes a range of physiologic responses including microarousals, delta and K-complex bursts, and variations in autonomic system. Whether patterns in slow-wave electroencephalographic activity and autonomic activation are primary forms of arousal response can be addressed by studying effects of total sleep deprivation. We therefore examined changes in arousal density during recovery sleep in healthy subjects. DESIGN: Participants spent 6 consecutive 24-hour periods in the laboratory. Nights 1 and 2 were baseline nights followed by 64-hour total sleep deprivation, then 2 consecutive recovery nights. SETTING: Sleep-deprivation protocol was conducted under laboratory conditions with continuous behavioral and electrophysiologic monitoring. PARTICIPANTS: Twelve drug-free men aged 27.4 +/- 7.9 years were studied. None reported sleepiness or altered sleep-wake cycle, and none had neurologic, psychiatric or sleep disorders. INTERVENTION: N/A. MEASUREMENTS AND RESULTS: Arousals were classified into 4 levels: microarousals, phases of transitory activation, and delta and K-complex bursts. Sleep deprivation induced changes in the density of considered arousals except phases of transitory activation, with a distinct pattern among the different types. The greatest change was found for microarousals, which showed a significant decrease in the first recovery night (P = .01), with return to baseline thereafter. A fall in K-complex and delta-burst density was noted in the first recovery night, not, however, reaching statistical significance. The phases of transitory activation rate were virtually unaffected throughout the experimental nights. CONCLUSIONS: We conclude that homeostatic sleep processes exert an inhibitory effect on arousal response from sleep with a significant effect only on the microarousal density. Decreased delta and K-complex burst rates, though not significant, support the hypothesis that they may be activating processes, probably modulated by factors independent from those implicated in cortical arousal. PMID- 15532201 TI - Complete or partial circadian re-entrainment improves performance, alertness, and mood during night-shift work. AB - STUDY OBJECTIVES: To assess performance, alertness, and mood during the night shift and subsequent daytime sleep in relation to the degree of re-alignment (re entrainment) of circadian rhythms with a night-work, day-sleep schedule. DESIGN: Subjects spent 5 consecutive night shifts (11:00 pm-7:00 am) in the lab and slept at home in darkened bedrooms (8:30 am-3:30 pm). Subjects were categorized by the degree of re-entrainment attained after the 5 night shifts. Completely re entrained: temperature minimum in the second half of daytime sleep; partially re entrained: temperature minimum in the first half of daytime sleep; not re entrained: temperature minimum did not delay enough to reach daytime sleep. SETTING: See above. PARTICIPANTS: Young healthy adults (n = 67) who were not shift workers. INTERVENTIONS: Included bright light during the night shifts, sunglasses worn outside, a fixed dark daytime sleep episode, and melatonin. The effects of various combinations of these interventions on circadian re entrainment were previously reported. Here we report how the degree of re entrainment affected daytime sleep and measures collected during the night shift. MEASUREMENTS AND RESULTS: Salivary melatonin was collected every 30 minutes in dim light (<20 lux) before and after the night shifts to determine the dim light melatonin onset, and the temperature minimum was estimated by adding a constant (7 hours) to the dim light melatonin onset. Subjects kept sleep logs, which were verified by actigraphy. The Neurobehavioral Assessment Battery was completed several times during each night shift. Baseline sleep schedules and circadian phase differed among the 3 re-entrainment groups, with later times resulting in more re-entrainment. The Neurobehavioral Assessment Battery showed that performance, sleepiness, and mood were better in the groups that re-entrained compared to the group that did not re-entrain, but there were no significant differences between the partial and complete re-entrainment groups. Subjects slept almost all of the allotted 7 hours during the day, and duration did not significantly differ among the re-entrainment groups. CONCLUSIONS: In young people, complete re-entrainment to the night-shift day-sleep schedule is not necessary to produce substantial benefits in neurobehavioral measures; partial re entrainment (delaying the temperature minimum into the beginning of daytime sleep) is sufficient. The group that did not re-entrain shows that a reasonable amount of daytime sleep is not enough to produce good neurobehavioral performance during the night shift. Therefore, some re-alignment of circadian rhythms is recommended. PMID- 15532202 TI - Different criteria of sleep latency and the effect of melatonin on sleep consolidation. AB - OBJECTIVES: Since there is no consensus definition of sleep onset, we studied different aspects of initial sleep periods in healthy volunteers taking melatonin. Two criteria for sleep latency were used: 10 minutes of uninterrupted sleep and 1.5 minutes of stage 1 sleep. PARTICIPANTS: Forty healthy male volunteers (mean age 28 +/- 5 years) were assigned to 2 groups: 30 ingested melatonin and 10 placebo. DESIGN: All volunteers underwent an initial polysomnogram (baseline) after a 1-night adaptation period. The next day, the placebo or a 10-mg dose of melatonin was administered for 28 days, 1 hour before sleep time, in double-blind fashion. The second polysomnogram was recorded on day 14. SETTING: Sleep laboratory RESULTS: Chronic melatonin administration led to a significant reduction in sleep latency, using only the criterion 10 minutes of uninterrupted sleep. This effect suggests that melatonin may have a hypnotic effect, and the use of melatonin may lead to better sleep consolidation. CONCLUSIONS: These results show differences that have clinical implications, since the criteria used to diagnose initial insomnia were based on sleep onset. PMID- 15532203 TI - Sleep quality and blood pressure dipping in normal adults. AB - OBJECTIVES: To investigate the relationship between sleep quality and nocturnal blood pressure dipping in normal subjects. We hypothesized that sleep quality correlates with dipping. DESIGN: Cross-sectional study. SETTING: Unattended polysomnography in the home followed by a 24-hour ambulatory blood pressure measurement. PATIENTS: Eighty-eight self-described normal subjects were evaluated; 26 were excluded due to an apnea-hypopnea index > or = 10. None were taking antihypertensive medications. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Subjects were divided into dippers and nondippers based on > or = 10% drop in nocturnal mean arterial pressure (MAP). Sleep-quality variables included total sleep time; sleep latency; percentage of stages 1, 2, 3, 4, and rapid-eye movement sleep; percentage of wake time after sleep onset (WASO); total arousal index; and sleep efficiency. Of the remaining 62 subjects, 17.7% were nondippers, and 7 were hypertensive. There was no difference in age, body mass index, apnea hypopnea index, blood pressure, or sleep quality between groups. Stage 4 sleep correlated significantly with dipping of diastolic blood pressure and MAP (r = 0.410 and 0.378, respectively, P < or = .002), and percentage of WASO was negatively correlated with dipping of diastolic blood pressure (r = -0.360, P = .004), suggesting that greater dipping was associated with better sleep quality. On multivariate analyses, Stage 4 sleep was independently associated with dipping of diastolic blood pressure (P = .034) after adjusting for screening MAP, percentage of WASO, total arousal index, and Stage 1 sleep. The same link was found between Stage 4 sleep and dipping of MAP (P = .05) after adjusting for screening MAP, age, sex, and body mass index. Repeat analyses excluding hypertensives yielded similar findings. CONCLUSION: Our data suggest that deeper and less-fragmented sleep is associated with more blood pressure dipping in normal subjects. PMID- 15532204 TI - Postural effects on pharyngeal protective reflex mechanisms. AB - STUDY OBJECTIVES: Pharyngeal muscle dilators are important in obstructive sleep apnea pathogenesis because the failure of protective reflexes involving these muscles yields pharyngeal collapse. Conflicting results exist in the literature regarding the responsiveness of these muscles during stable non-rapid eye movement sleep. However, variations in posture in previous studies may have influenced these findings. We hypothesized that tongue protruder muscles are maximally responsive to negative pressure pulses during supine sleep, when posterior tongue displacement yields pharyngeal occlusion. DESIGN: We studied all subjects in the supine and lateral postures during wakefulness and stable non rapid eye movement sleep by measuring genioglossus and tensor palatini electromyograms during basal breathing and following negative pressure pulses. SETTING: Upper-airway physiology laboratory of Sleep Medicine Division, Brigham and Women's Hospital. SUBJECTS/PARTICIPANTS: 17 normal subjects. MEASUREMENTS AND RESULTS: We observed an increase in genioglossal responsiveness to negative pressure pulses in sleep as compared to wakefulness in supine subjects (3.9 percentage of maximum [%max] +/- 1.1 vs 4.4 %max +/- 1.0) but a decrease in the lateral decubitus position (4.1 %max +/- 1.0 vs 1.5 %max +/- 0.4), the interaction effect being significant. Despite this augmented reflex, collapsibility, as measured during negative pressure pulses, increased more while subjects were in the supine position as compared with the lateral decubitus position. While the interaction between wake-sleep state and position was also significant for the tensor palatini, the effect was weaker than for genioglossus, although, for tensor palatini, baseline activity was markedly reduced during non rapid eye movement sleep as compared with wakefulness. CONCLUSION: We conclude that body posture does have an important impact on genioglossal responsiveness to negative pressure pulses during non-rapid eye movement sleep. We speculate that this mechanism works to prevent pharyngeal occlusion when the upper airway is most vulnerable to collapse eg, during supine sleep. PMID- 15532205 TI - Sex influences endothelial function in sleep-disordered breathing. AB - BACKGROUND: The bases for the association between sleep-disordered breathing (SDB) and cardiovascular disease are poorly understood. Endothelial dysfunction, assessed with brachial artery ultrasonography, may predict cardiovascular risk and represent preclinical vascular disease. We determined whether flow-mediated dilation (FMD) and peak blood flow (PBF) increase after cuff occlusion is altered with SDB. METHODS: 193 participants (58% women) in a cohort study were studied with overnight polysomnography and subsequent brachial artery ultrasonography. SDB was quantified using the apnea-hypopnea index (AHI) and indexes of overnight desaturation and arousal frequency. Two-dimensional and Doppler-velocity measurements of the brachial artery were obtained at baseline and after 5 minutes of upper-arm cuff occlusion. FMD and PBF were defined as the percentage changes from baseline in brachial artery diameter and flow, respectively. RESULTS: In the entire sample, the AHI was inversely associated with both FMD (r = -0.30, P < .001) and PBF (r = -0.20, P < .001). However, sex-stratified univariate analyses showed that these relationships were exclusive to women. Specifically, FMD was significantly lower in women with an AHI > or = 15 than in women with lower AHI scores (P < .005), with no relationship between AHI and FMD in men. Additionally, PBF decreased significantly with increasing AHI (r = -0.29, P = .010) in women alone. Statistical modeling, adjusting for body mass index, age, and other covariates, similarly showed that SDB severity significantly influenced FMD and PBF, with significant interactions between sex and AHI, reinforcing that the associations between SDB severity and endothelial function were stronger in women than in men. CONCLUSIONS: Moderate levels of SBD are associated with impaired conduit and resistance endothelial function in women. Women with SDB may be more vulnerable to early SDB-related cardiovascular disease than are men. PMID- 15532206 TI - The socioeconomic impact of narcolepsy. AB - STUDY OBJECTIVE: To evaluate the economic burden of patients suffering from narcolepsy. DESIGN: Cross-sectional study. PARTICIPANTS: 75 narcoleptic patients diagnosed at the Hephata Klinik, Germany, who met the International Classification of Sleep Disorders criteria for narcolepsy. MEASUREMENTS: A standardized telephone interview was used to inquire about the disease and its burden. In addition, health-related quality of life scales (SF-36 and EQ-5D) were mailed to the patients. Complete data were available for 75 patients. Direct and indirect costs were calculated from the societal perspective. All costs (mean +/- SD) were calculated in 2002 Euros (Euros). RESULTS: Total annual costs were Euros 14,790 +/- 16,180 (US-dollars 15,410) per patient. Direct costs were Euros 3,180 +/- 3,540 (US-dollars 3,310), which consisted of hospital costs (Euros 1,210 +/- 2,210; US-dollars 1,260), drug costs (narcolepsy medication: Euros 1,020 +/- 1,420; US-dollars 1,060), ambulatory care (Euros 90 +/- 90; US-dollars 40), and ambulatory diagnostics (Euros 20 +/- 30; US-dollars 20). More than 50% of the drug costs were due to the newer wake-promoting drugs. Total annual indirect costs amounted to Euros 11,390 +/- 15,740 (US-dollars 11,860) per patient and were mainly attributable to early retirement due to the disease. Narcolepsy was listed as being the cause of unemployment by 32 patients. In patients with sleep attacks, there was a significant correlation between early retirement and higher indirect costs compared to patients without these attacks. CONCLUSIONS: Narcolepsy causes a high socioeconomic burden, which is comparable to other chronic neurologic diseases, Indirect costs are considerably higher than direct costs. Measures should be taken to increase public awareness of this disease. Appropriate treatment must be provided in order to increase quality of life and to assist patients continuing in working life. PMID- 15532207 TI - Health-related quality of life and depressive symptoms in children with suspected sleep-disordered breathing. AB - STUDY OBJECTIVES: Snoring and sleep-disordered breathing (SDB) are highly prevalent among children. The increasing trends of obesity in the pediatric population further predict an exacerbation of this public health problem. However, the impact of SDB on mood and quality of life and the confounder effect of obesity in this setting are unclear. DESIGN: We studied a group of 85 clinically referred children, aged 8 to 12 years, with snoring and suspected SDB and 35 asymptomatic children (controls). All children completed validated questionnaires for the presence of depression (Children's Depression Inventory) and for health-related quality of life (PedsQL), and parents completed the Parent Report version of the PedsQL. Children referred to the Sleep Medicine Center for sleep-disordered breathing were further subdivided according to their body mass index > 95% for age and sex (n = 44) and with BMI < 95% for age and sex as ClinN1 (n = 41). RESULTS: Parentally reported quality of life and physical health differed between obese and nonobese children. However, both groups with SDB had more-impaired quality of life and depressive symptoms than did controls. CONCLUSIONS: Children with suspected SDB, regardless of the severity of apnea hypopnea index or the presence of obesity, had more impairments in quality of life and depressive symptoms than did children who did not snore. PMID- 15532208 TI - Reliability of scoring arousals in normal children and children with obstructive sleep apnea syndrome. AB - STUDY OBJECTIVES: Scoring of arousals in children is based on an extension of adult criteria, as defined by the American Sleep Disorders Association (ASDA). By this, a minimum duration of 3 seconds is required. A few recent studies utilized modified criteria for the study of children, with durations as short as 1 second. However, the validity and reliability of scoring these shorter arousals have never been verified. Based on studies in adults, we hypothesized that interscorer agreement for scoring arousals shorter than 3 seconds was poor. DESIGN: Retrospective review of polysomnograms by 2 experienced sleep practitioners who independently scored arousals according to the ASDA 3-second criteria and modified duration criteria of 1 and 2 seconds. SETTING: Academic hospital. PATIENTS OR PARTICIPANTS: 20 polysomnographic studies from children aged 3 to 8 years with mild to severe obstructive sleep apnea syndrome, and 16 polysomnographic studies from normal children. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: The intraclass correlation coefficient for scoring ASDA arousals was 0.90 (95% confidence interval: 0.81-0.95), indicating excellent interscorer agreement. The intraclass correlation coefficient for scoring modified 1-second and 2-second arousals were 0.35 (95% confidence interval: 0.02-0.61) and 0.42 (95% confidence interval: 0.12-0.65) respectively, indicating poor to fair interscorer agreement. Furthermore, modified 1-second and 2-second arousals accounted for less than 15% of all arousals scored. CONCLUSIONS: We conclude that there is much poorer interscorer agreement for scoring arousals shorter than 3 seconds, when compared to the standard ASDA criteria. We propose that scoring of arousals in children should follow the standard ASDA criteria. PMID- 15532209 TI - Effects of maternal separation on baseline sleep and cold stress-induced sleep rebound in adult Wistar rats. AB - STUDY OBJECTIVES: To evaluate the influence of early life environments on basal and cold stress-induced sleep patterns in rats. DESIGN: The design was a 3 (Groups [control, early handling, maternal separation]) x 2 (Situations [basal, poststress]) x 11 (Time-blocks) factorial design. From postnatal days 2 to 14, whole litters were either submitted to early handling (15 minutes per day away from the mother) or maternal separation (180 minutes per day away from the mother). At 75 to 90 days of age, sleep was recorded for 22 hours (beginning at 9:00 AM) before and after 1 hour of cold stress (4 degrees C). SUBJECTS: Wistar male rats (n = 7-10 animals per group). MEASUREMENTS AND RESULTS: Sleep was analyzed in blocks of 2 hours, in 30-second epochs, separately in the daytime and nighttime sleep recordings. Maternally separated rats exhibited more paradoxical sleep at baseline, compared to both control and early-handled rats. In the first 2 hours following the end of cold stress, all groups showed a decrease in paradoxical sleep, whereas slow-wave sleep was reduced only in the control group. The highest corticosterone plasma concentration was observed immediately after stress. Sleep rebound after stress was equally manifested in all groups in the dark part of the light-dark cycle. CONCLUSIONS: Maternal separation during early infancy resulted in permanent changes of the sleep architecture reflected by augmented time spent in paradoxical sleep. Although these findings were not expected in light of the literature, they emphasize the importance of the early familiar environment on future behavior of rats. PMID- 15532210 TI - Children with obstructive sleep-disordered breathing generate high-frequency inspiratory sounds during sleep. AB - STUDY OBJECTIVES: We observed that some children with adenotonsillar hypertrophy and obstructive sleep-disordered breathing (SDB) make high-frequency inspiratory sounds (HFIS) during sleep. Our objective was to determine whether HFIS occur in most children with obstructive SDB and adenotonsillar hypertrophy and whether adenotonsillectomy reduces HFIS. DESIGN: Prospective consecutive-entry trial. SETTING: Sleep laboratory. PARTICIPANTS: Twenty-six children between 6 and 12 years of age with adenotonsillar hypertrophy suspected of having obstructive SDB. MEASUREMENTS AND RESULTS: We performed polysomnography and measured sounds during sleep with a microphone suspended above the bed. Sounds were recorded on a computer at 44 kHz, analyzed with fast Fourier transformation for frequency content. HFIS were sounds occurring during an inspiration with frequencies greater than 2 kHz. HFIS were different from the low-frequency (< 2 kHz) sounds described in snoring adults. HFIS usually occurred in consecutive breaths, occasionally exceeding 100. We counted the number of HFIS that occurred per hour of sleep. Children who made more HFIS had more obstructive SDB than did those who did not make the HFIS, and there was a significant positive correlation between the number of HFIS and the obstructive apnea-hypopnea index. Children with more than 3 apneas and hypopneas per hour of sleep all made at least 10 HFIS per hour, and all children who had more than 10 HFIS per hour had obstructive apnea hypopnea index values greater than 1. Children with adenotonsillar hypertrophy made more HFIS than did those children whose tonsils and adenoids had been removed. CONCLUSIONS: HFIS may be a marker of disturbed breathing during sleep in children with adenotonsillar hypertrophy. PMID- 15532211 TI - Who reports insomnia? Relationships with age, sex, ethnicity, and socioeconomic deprivation. AB - STUDY OBJECTIVES: To investigate the prevalence of self-reported insomnia symptoms among Maori (indigenous people) and non-Maori adults in the general population of New Zealand. To explore the possible links between insomnia symptoms and ethnicity, gender, age, employment status and socio-economic deprivation. DESIGN: Mail-out survey to a stratified random sample of 4,000 people aged 20 to 59 years, selected from the electoral roll. SETTING: Nationwide survey of New Zealand adults (72.5% response rate). PARTICIPANTS: The sample design aimed for equal numbers of Maori and non-Maori participants, men and women, and participants in each decade of age. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Population prevalence estimates indicated that self-reported insomnia symptoms and sleeping problems were higher among Maori than non-Maori. Multiple logistic regression analyses identified unemployment and socioeconomic deprivation as being strongly associated to all insomnia symptoms and to reporting a sleeping problem lasting more than 6 months. CONCLUSIONS: Socioeconomic factors and ethnicity are significant independent predictors of reported insomnia symptoms. This finding has important implications for the provision of treatment services to those most in need. PMID- 15532212 TI - Comparison of limited monitoring using a nasal-cannula flow signal to full polysomnography in sleep-disordered breathing. AB - STUDY OBJECTIVES: Evaluate the utility of overnight monitoring limited to nasal cannula airflow and oximetry in the diagnosis of obstructive sleep apnea-hypopnea syndrome (OSAHS). DESIGN: Prospective randomized study, blinded analysis. SETTING: Sleep disorder center, academic institution. PARTICIPANTS: 56 patients with suspected OSAHS, 10 normal volunteers. MEASUREMENTS AND RESULTS: In laboratory full nocturnal polysomnography (NPSG) and unattended ambulatory study with monitoring of only airflow and oximetry performed in randomized order. Obstructive respiratory events were scored on the full NPSG while visualizing all signals and then rescored on the full NPSG and on the ambulatory study while visualizing only airflow and oximetry signals. Respiratory disturbance indexes (RDI) for the limited studies (RDIFlowNPSG and RDIFlowAmbulatory) were calculated as the sum of the apneas and hypopneas (defined using airflow amplitude and O2 desaturation) divided by the valid flow-signal time. The reference RDIFullNPSG was calculated from the sum of the apneas and hypopneas (defined using flow amplitude, O2 desaturation and electroencephalographic arousal) identified on the full NPSG divided by the total sleep time. RDIFullNPSG was greater than RDIFlowNPSG (bias = 5.6 events per hour) and RDIFlowAmbulatory (bias = 10.9 events per hour), but the differences were mainly in subjects with an RDI > 40 events per hour. The diagnostic sensitivity and specificity for the diagnosis of OSAHS using a cutoff of 18 events per hour were 96% and 93% using the flow signal from the NPSG and 88% and 92% using the flow signal from the ambulatory study performed on a separate night. CONCLUSIONS: In subjects with OSAHS, analysis of the flow signal from a nasal cannula can provide an RDI similar to that obtained in a full NPSG. PMID- 15532213 TI - What keeps us awake: the neuropharmacology of stimulants and wakefulness promoting medications. AB - Numerous studies dissecting the basic mechanisms that control sleep regulation have led to considerable improvement in our knowledge of sleep disorders. It is now well accepted that transitions between sleep and wakefulness are regulated by complex neurobiologic mechanisms, which, ultimately, can be delineated as oscillations between two opponent processes, one promoting sleep and the other promoting wakefulness. The role of several neurotransmitter or neuromodulator systems, including noradrenergic, serotonergic, cholinergic, adenosinergic, and histaminergic systems and, more recently, the hypocretin/orexin and dopamine systems, has been clearly established. Amphetamine-like stimulants are known to increase wakefulness by blocking dopamine reuptake, by stimulating dopamine release, or by both mechanisms. Modafinil may increase wakefulness through activation of noradrenergic and dopaminergic systems, possibly through interaction with the hypocretin/orexin system. Caffeine inhibits adenosinergic receptors, which in turn can produce activation via interaction with GABAergic and dopaminergic neurotransmission. Nicotine enhances acetylcholine neurotransmission in the basal forebrain and dopamine release. Understanding the exact role of the hypocretin/orexin and dopamine systems in the physiology and pharmacology of sleep-wake regulation may reveal new insights into current and future wakefulness-promoting drugs. PMID- 15532214 TI - Delayed sleep phase syndrome: pathophysiology and treatment options. AB - This paper presents a comprehensive review of delayed sleep phase syndrome (DSPS), a circadian-rhythm sleep disorder thought to result from the endogenous circadian pacemaker being "stuck" at a later-than-normal phase, relative to the desired sleep-wake schedule. A full understanding of this disorder is best appreciated from the context of shared modulation of sleep and wakefulness via sleep homeostatic and circadian systems. Typically emerging during adolescence, DSPS comes to clinical attention much less often than prevalence estimates would suggest, perhaps due to underrecognition by clinicians and misattribution of symptoms. Several treatment modalities have been suggested, including phototherapy, chronotherapy, and exogenous melatonin administration. However, caution is raised for the reason that more than 20 years after its initial description in the literature, the basic pathophysiology of DSPS remains poorly understood, as observed in the 2003 National Sleep Disorders Research Plan. Challenges for future research include elucidating the exact sleep homeostatic and circadian contributions to the disorder, improving the objective verification of this diagnosis instead of relying only on self-report information, and conducting treatment research aimed at determining efficacy, effectiveness, and mechanism or mechanisms of action. PMID- 15532215 TI - SLEEP year in review 2003--insomnia, sleep in psychiatric disorders, periodic limb movement disorder/restless legs syndrome, sleep in medical disorders. PMID- 15532216 TI - A year in review--basic science, narcolepsy, and sleep in neurologic diseases. PMID- 15532217 TI - REM-sleep changes in children with attention-deficit/hyperactivity disorder: methodologic and neurobiologic considerations. PMID- 15532218 TI - An early description of REM sleep behavior disorder. PMID- 15532219 TI - Ideal practice versus best workable practice. PMID- 15532220 TI - Improving measurement of CPAP self-efficacy. PMID- 15532221 TI - Gastro-oesophageal reflux disease: a review of clinical features, investigations and recent trends in management. AB - BACKGROUND: Gastro-oesophageal reflux disease, though a common digestive disorder worldwide, is scarcely reported with very sparse literature in oursetting. AIM: To review the clinical features, investigations and recent developments in the management of gastro-oesophageal reflux disease. METHODS: We retrieved publications from local and international journals. We also searched Medline particularly for local references. Other sources of our data include Up-to-date in Medicine and standard texts in medicine and pathology. RESULTS: Local literature is generally lacking; while the classical symptoms of gastro oesophageal reflux disease are heartburn, dysphagia and acid regurgitation, clinical examination is usually silent except when there are complications. There is no gold standard investigation but oesophageal pH monitoring, Bernstein test, oesophagoscopy and oesophageal manometry have been established to be useful in the evaluation of affected patients. Radiologic investigations, though insensitive in the diagnosis, are invaluable in diagnosing complications. Response to 14-day treatment with omeprazole (20 mg daily) has now been established to have a high sensitivity and specificity in diagnosing the condition. Management strategies include lifestyle modification, medical and surgical therapies. Proton pump inhibitors are now recognized as first line therapy in management. CONCLUSION: The prevalence and behaviour of this condition in Nigeria is not known and local references are scanty. This review article focused on the worldwide epidemiology, pathogenesis and recent trends in management to stimulate our interest in this area. PMID- 15532222 TI - Perioperative management of the diabetic patient. AB - BACKGROUND: The prevalence of diabetes mellitus is increasing worldwide and many more diabetics are requiring surgical intervention for various reasons. There is no general agreement on a satisfactory routine for managing diabetics during surgery while many aspects of its care also remain controversial. METHOD: The broad medical literature on the perioperative management of the diabetic surgical patient was reviewed. Meta analysis, review articles, original articles and expert recommendations were identified on a search of the pubmed. RESULT: This review addresses the important aspects of pre-, intra-and post-operative management of the diabetic surgical patient and highlights recent advances on the subject. CONCLUSION: Different types of regimens are now available for managing the diabetic surgical patient. Optimal methods such as intravenous insulin infusions, though more laborious appear to be most popular. Clinical judgement remains a vital component of good perioperative management of patients with diabetes mellitus. PMID- 15532223 TI - Recurrent respiratory papillomatosis in Ibadan, Nigeria. AB - BACKGROUND: Recurrent respiratory papilomatosis (RRP), the most common benign neoplasm of the larynx, usually results in upper airway obstruction that is most often misdiagnosed as lower airway disease in children. The prevalence of RRP and various aspects of its clinical course are still unknown with a dearth of literature on this in Nigeria. This study aims at presenting the clinical profile of RRP as seen in the University College Hospital, Ibadan, Nigeria. METHODOLOGY: A retrospective study of 43 cases of histologically confirmed RRP over a fifteen year period in the Otorhinolaryngology Department of the University College Hospital Ibadan. RESULTS: There were 28 (65.1%) males and 15 (34.9%) females with mean age of 8.7 years (range 2-23 years). Thirty-two patients (74.4%) were between 110 years with the highest incidence occurring at the 6-10 year age group (39.5%). Forty-one patients (95.3%) belonged to low social classes V and IV. Thirty-one patients (72%) presented within 1 year of onset of symptoms. All the patients were hoarse with 70% of those dyspnoeic at presentation having emergency tracheostomy. The lesions involved the vocal cords and anterior commissure mostly. There were no recurrences in 23 patients (53.5%) while twenty patients (46.5%) had 2 to 5 recurrences. CONCLUSION: Sequelae of RRP in Children may be grave, thus a high index of suspicion is needed in a patient with progressive voice change exceeding six weeks and unresponsive to standard medical therapy. PMID- 15532224 TI - A review of appendicectomy in Sokoto, North-western Nigeria. AB - BACKGROUND: Acute appendicitis is a common surgical emergency in developed countries. There is a strorng evidence of a rising trend in the developing world. METHODS: We present a retrospective review of appendicectomy as performed at Usmanu Danfodiyo University Teaching Hospital, Sokoto over a six year period (January 1997 to December, 2002) RESULTS: The diagnosis of appendicitis was made in 38.9% of surgical abdominal emergencies. The peak age group was 20 to 29 years with a male to female ratio of 1.4:1. The negative appendicectomy rate was 15.9% There was associated Schistosoma haematobium in 4.44% of cases. Appendicectomy had a morbidity rate of 13.3% and a mortality of 0.9%. CONCLUSIONS: Appendicitis was the commonest surgical emergency in Sokoto. The negative appendicectomy, morbidity and mortality rates were within the range widely quoted in the literature. The need to avoid negative exploration is imperative in this poor community. PMID- 15532225 TI - Complications of intrauterine contraceptive device (IUD) use at the University of Ilorin Teaching Hospital, Ilorin. AB - BACKGROUND: The use of Intrauterine Contraceptive Device (IUD) has increased steadily worldwide over the years. METHODS: This is a retrospective study of 602 women who had IUD insertions at the Family Planning Unit of the University of Ilorin Teaching Hospital between January- December 1995 and the study was undertaken to study the complications associated with the IUD use at the University of Ilorin Teaching Hospital. RESULTS: There was 85.55% continuation rate after a year, 81.06% of patients were without complications, 0.33% had Pelvic Inflammatory Disease (PID) and no perforation was reported. Expulsion was also low. Mean duration of use was 14.06 +/- 4.46 months. There was no statistical difference in timing of insertion in those with or without complications (p = 0.781); however, a significant difference becomes noticeable in the duration of use of the IUD in the two groups (p = 0.001). IUD was acceptable to most patients (85.5%). There was no significant difference with respect to level of education (p = 0.064) and type of previous contraception practised (p = 0.022) in those with or without complications. IUD complications were commonest within the first 3 months of insertion; involving a third of those lost to follow-up compared with about 15% of those not lost to follow-up (p = 0.001). CONCLUSIONS: At the University of florin Teaching Hospital, Ilorin, IUD use is safe and effective with minimal complications. PMID- 15532226 TI - Prevalence and medicalization of female genital mutilation. AB - BACKGROUND: Most traditional practices in Africa, sustained and fuelled by underdevelopment, ignorance, illiteracy, poverty and tenacious customs, have been on the decline except for female genital mutilation (FGM). AIM: To ascertain the prevalence and operators of female genital mutilation (FGM). METHOD: A cross sectional study was carried out in a semi-urban town in Rivers State of Nigeria between January and April 1988, utilizing structured questionnaires which were randomly distributed. The response of 600 respondents was analyzed. RESULTS: Prevalence of FGM amongst the study population was 53.2%. Medical doctors were the most mentioned operators (34.5%) followed by traditional birth attendants (33.8%). Control of excess female sexual desires (58.2%) and promotion of culture (39.5%) were the major reasons adduced for this practice. CONCLUSION: The enforcement of the World Health Organization (WHO) legislation against medicalization of this practice is advocated. Training and retraining of traditional birth attendants with emphases on eradication of harmful cultural practices will help. PMID- 15532227 TI - Knowledge, attitude and practice of female genital cutting among antenatal patients in Aminu Kano Teaching Hospital, Kano. AB - BACKGROUND: Despite the widespread practice of FGC, not much attention had been given to it until recently. The attitude of expectant mothers towards it is crucial in sustaining it. OBJECTIVE: To assess knowledge, attitude and practice of female genital cutting among antenatal patients in Aminu Kano Teaching hospital in northern Nigeria METHODS: A cross-sectional study was conducted on 210 antenatal patients seen at Aminu Kano Teaching Hospital from February to March 2003. A structured interviewer-administered questionnaire was used to collect information about socio-demographic characteristics, knowledge, attitude and practice of female genital cutting. RESULTS: Majority (91.4%) of the respondents have heard about FGC. Clitoridectomy and infibulation were known by 36.2% and 5.2% of the respondents respectively. Sixty and 5.2% of respondents knew that HIV/AIDS and Hepatitis could be transmitted by methods used in FGC. Other complications mentioned include sexual dissatisfaction (25.7%) and difficult delivery (8.6%). Furthermore, 16% of respondents wished FGC would continue mainly due to cultural or religious reasons, this opinion was associated with low (20.5%) level of education. Opponents of FGC gave medical complications as their main reason. Prevalence of circumcision was 23.3% and the most common type (36.7%) was clitoridectomy. CONCLUSION: Though awareness about FGC has improved, and attitude towards it appears generally negative, the practice still persists in the north, though at a lower level than the Southeast and Southwest. It was therefore recommended that intensive campaign and health education among mothers and the general public, as well as enforcement of legislation are needed. PMID- 15532228 TI - Abdominal massage: another cause of maternal mortality. AB - BACKGROUND: Abdominal massage is an age long practice among obstetric and non obstetric patients in different parts of the riverine areas of Southern Nigeria. AIM: To highlight the consequences associated with this practice in pregnancy and labour even among literate clients. METHODS: Two hundred and eighty four pregnant women who presented as obstetric emergencies in pregnancy and labour at the University of Port Harcourt Teaching Hospital from 1st May 1988 to 31st July 1998 were studied. RESULTS: Forty two (14.79%) of the 284 patients had abdominal massage. Maternal mortality rate was 4.76% while the perinatal mortality rate was 14.29%. Twenty four of the patients were literate though 15 of them only had primary education. Most of the patients were unbooked (76.19%). The complication of pregnancy noted were cephalo pelvic disproportion /obstructed labour (28. 57%) and abruption placenta (23.80%); placenta previa (9.52%); retained placenta (4.76%) abortion / preterm labour (19.04%); genital injuries and tears (7.14%); perinatal death (14.28%); uterine rupture (9.52%); maternal mortality (4.76%). CONCLUSION: Abdominal massage is a silent killer, which has added to matemal and perinatal mortality and morbidity in this region. Ignorance, illiteracy and poverty have sustained this practice, while underdevelopment, inaccessibility and ill-equipped medical facilities have kept this traditional practice flourishing even amongst the educated group. The time to stop this form of avoidable losses is now. This can be achieved through public health approach, mass literacy, even distribution of affordable healthcare services, improved social services and transportation, robust economy and a grass root involvement of rural programmes aimed at eradicating harmful traditional practice. PMID- 15532229 TI - ABO and rhesus antigens in a cosmopolitan Nigeria population. AB - BACKGROUND: Port Harcourt is a cosmopolitan city consisting of several ethnic groupings such as Ikwerre, Ijaw, Igbo, Ogonis, Efik-Ibibio, Edo, Yoruba, Hausa and foreign nationals. ABO and Rhesus D antigens were screened in this cross sectional study with the aim of generating data that would assist in the running of an efficient blood transfusion service for a cosmopolitan city as Port Harcourt. METHOD: Blood donors were sampled and screened for ABO and Rhesus D antigens at three Health facilities within Port Harcourt: University of Port Harcourt Teaching Hospital, Braithwaite Memorial Hospital and Orogbum Health centre. RESULTS: A total of 936 blood donors were tested in this study. The results of the ABO screening shows that blood group O was the highest with 527 (56.30%) followed by blood group A, B and lastly AB with 212 (22.65%), 178 (19.02%) and 18(2.10%) respectively. The highest contribution to blood group O was from the Ibos with 220 (23.50%) while the Ijaws gave the highest contribution of Rhesus "D" antigen with 370 (39.53%), closely followed by the Igbos with 334 (0.43%). Rhesus negativity values in this study was 7.26% of which the highest contributors were also the Ijaws with 33 (3.53%) and Igbos with 27(2.89%). CONCLUSION: The increased demand for safe blood calls for an efficient Blood, Transfusion Service at the local, state and national levels. It is hoped that the data generated in this study would assist in the planning and establishment of a functional Blood service that would not only meet the ever increasing demand for blood products, but also play a vital role in the control of HIV/AIDS and . Hepatitis B global scourge. PMID- 15532230 TI - Socio-demographic characteristics and risk factors among HIV/AIDS patients in Kano, Northern Nigeria. AB - BACKGROUND: There has been no description of the characteristics of HIV/AIDS patients that are referred to this antiretroviral treatment centre. The information could be useful in planning for prevention, care and support of people living with HIV/AIDS (PLWHA) and people affected by AIDS (PABA). OBJECTIVES: To describe the socio-demographic characteristics and identify risk factors among HIV/AIDS patients presenting to Aminu Kano Teaching Hospital. METHODS: Analysis of a prospectively collected data on 205 consecutive patients presenting to Aminu Kano Teaching Hospital from May 2002 to June 2003. RESULTS: Out of the 205 HIV positive patients seen, there were 112 (54.6%) males and 93 (45.4%) females giving a sex ratio of 1.2:1. The age range was 18 to 61 years with an overall mean S.D of 33.7 +/- [corrected] 8 years. The mean for males was 37.3 +/- [corrected] 7.8 years compared to 29.56 years for females. This difference was statistically significant (t = 8.1 DF = 201 P < 0.001). About 38.5% of the patients had secondary school education, 36.1% had tertiary education and 10.2% had primary education. Half of the patients were Hausas, 18.5% Igbos, 4.9% Idomas and 3.4% Igalas. Sixty percent were Muslims, the rest were Christians. About 64.0% were married, 23.0% were single and 11.0 % were divorced. Only 17.0% reported having multiple sexual partners. Eighteen percent claimed they use condoms always, 67.8% occasionally and 8.3% never. Twenty percent of the patients had previous blood transfusion. None reported intravenous drug abuse or homosexuality. CONCLUSION: The need for a sustained multi-sectoral preventive effort and the scaling up of treatment and community care initiatives were stressed. PMID- 15532231 TI - The influence of pre- and post-partum factors on the time of contact between mother and her new-born after vaginal delivery. AB - BACKGROUND: The aim of this report is to determine the pre-partum and post-partum factors that may influence the time interval from vaginal delivery to first skin to-skin contact between the mother and her newborn at the University of Port Harcourt Teaching Hospital. METHOD: A prospective study of 250 consecutive healthy mother-infant pairs. Information on pre- and post-partum factors was obtained from interview of mothers, direct observation and medical records. The proportion of mothers who made skin-to-skin contact with their babies within 30 minutes of delivery (early contact) was compared with those who did not (late contact) in the various pre- and post-partum groups. RESULTS: There was early skin-to-skin contact between mother and child in ninety-six (38.4%) of the 250 mother-infant pairs. Age of mother below 25 years, primiparity, labour less than 12 hours and late repair of episiotomy were associated with early skin-to-skin contact between mother and child. Early performance of routine cleaning and taking measurements of newborn was significantly associated with late skin-to skin contact with mother. A significantly high proportion of mothers who were assisted to breastfeed had early contact with their babies compared with those who did not have any such assistance (48.8% vs. 17.1%). Ninety-six (39.7%) of the 242 mothers who received information on breastfeeding had early contact compared with none of the 8 mothers who did not have breastfeeding education. The mother's marital status and level of education did not show any statistically significant association with time of mother skin-to-skin contact with baby. CONCLUSION/RECOMMENDATION: Post-partum labour ward practices (cleaning and measurement of baby, assistance received by mother on breastfeeding and timing of repair of episiotomy) are the most important factors found to affect time of first contact between mother and child. It is recommended that immediate contact between newborn and mother should take precedence over hospital routines. PMID- 15532232 TI - Community participation in health and development. AB - BACKGROUND: This paper critically appraises the importance of community participation in health and development and the methods of involving them. It also emphasizes the role of every stakeholder in health and development. METHODS: These involved the review of some literature on this subject and examination of issues arising from the review. RESULTS: Community participation in health offers various advantages in health care and development among which are helping communities to develop problem solving skills, making them to take responsibility for their health and welfare, ensuring that the need and problems of the community are adequately addressed, ensuring that the strategies and methods used are culturally and socially appropriate or acceptable and finally it enhances sustainability. It is the responsibility of the government at various level, non governmental organizations, international health agencies and health care programme planners and providers to help the community to organize themselves and be involved in their health care and development. Them should be well established or institutionalized framework of making sure that people are consulted, persuaded, and given responsibility in decision making under technical and professional guidance of health care professionals. CONCLUSION: Plans must not be imposed or policies formulated without involving the community in all matters concerning health and development. PMID- 15532233 TI - Ethical issues in HIV/AIDS infections. AB - BACKGROUND: Human relationships are guided by codes of ethics, written or otherwise. The reality and the place of universal standards of ethics may be argued but local variations and interpretations of these universal standards do occur. METHOD: A search of some relevant articles on ethical issues on HIV/AIDS was undertaken through the medline and standard texts. RESULTS: Unlike any other disease HIV/AIDS infection has resulted in large amount of legal/ethical commentaries. For the most parts of the world HIV/AIDS issues have produced metamorphosing legal/ethical issues notably, the concern about confidentiality of medical information, informed consent for medical procedures/HIV testing, discrimination/stigmatisation of people living with HIV/AIDS, partner notification, the right of the patient to refuse treatment and the way counselling should be done. Clinicians in developing countries have had to rely on self-regulations in making ethical judgements or at least attempt to conform to poorly enforced ethical standards in clinical practice established by their institutions. CONCLUSION: Universally, recognized human rights standard should guide policy makers in formulating the direction and content of HIV related policy and should form an integral part of all aspects of national and local responses to HIV/AIDS. This paper therefore discusses ethical/legal issues presented by HIV/AIDS and how a variety of state, and federal laws and various institutions have attempted to address them. PMID- 15532234 TI - Designing a health education programme for the prevention and control of diarrhoea diseases at primary health care (PHC) level for local government (LGA) workers. AB - BACKGROUND: Diarrhoea diseases are a major cause of morbidity and mortality in developing countries especially among children. Diarrhoea diseases have [corrected] been linked to specific behavioural practices and host factors that increase susceptibility such as failure to breastfeed until at least one year of age, malnutrition and immunodeficiency state e.g. AIDS. The objective of this paper is to design a training method for the prevention and control of diarrhoea diseases for Local Government (LGA) workers. METHODS: This is a descriptive programme design for training highlighting the various steps essential for training on the prevention and control of diarrhoea diseases to LGA workers at the Primary Health Care (PHC) level. RESULTS: Programme design for the purpose of improving skills, knowledge and practices involves planning, implementation and evaluation. Planning involves identifying training needs by the collection and analysis of information related to specific need of each LGA worker at PHC level, setting training objectives and identifying the content of training (curriculum design). Implementation is the use of training methods such as methods of providing information, method of providing examples and methods of promoting practices. Evaluation involves the assessment of the immediate outcome of training and impact of training program. CONCLUSION: This training is important because every health or health-related worker who is in close contact with the people has the potential to influence the knowledge, attitude and health practices of the people with whom he or she works in the prevention and control of diarrhoea diseases. PMID- 15532235 TI - Spontaneous midtrimester uterine rupture: a case report. AB - BACKGROUND: Spontaneous mid trimester rupture of the uterus is uncommon. AIM: To report a case of spontaneous mid trimester uterine rupture in a patient with a previous caesarean section scar. METHOD: A review of the case record of a patient managed for spontaneous mid-trimester uterine rupture and the relevant literature. RESULTS: A 30-year old unbooked gravida 6, para 1(+4) house wife with a previous caesarean section scar presented with features of an acute surgical abdomen with hypovolaemic shock at 24 weeks gestation. She had an emergency laparotomy at which she was noted to have a uterine rupture. She had a repair of the uterus and bilateral tubal ligation. Her post operative state was uneventful. CONCLUSION: There should be a high index of suspicion of uterine rupture in a gravid woman with a previous uterine scar presenting with abdominal pain and shock. PMID- 15532236 TI - Physical and psychological sequelae of female genital mutilation: a case report. AB - BACKGROUND: One harmful traditional practice that has resisted change in many African countries is female genital mutilation (FGM), otherwise known as female circumcision. This is usually associated with many complications. This report highlights a combination of physical and psychological sequelae associated with female circumcision. METHOD: Case-note of a patient managed for complications of female genital mutilation was used with a review of the relevant literature. RESULT: A 17-year old, married, illiterate farmer who had complete labial fusion following circumcision is presented. She had a total of six failed repairs, with psychological trauma, prior to her presentation in this hospital. A successful vulval reconstructive surgery was preformed and she was able to start a normal life with her husband. CONCLUSION: FGM carries very high morbidity. All supporters [corrected] of reproductive health should initiate programmes to stop this harmful and dehumanizing practice. PMID- 15532237 TI - Choledochal cyst: a case report and review of literature. AB - BACKGROUND: We present our first experience in managing a case of choledochal cyst in a two and half year old girl highlighting the difficulties in the management together with a review of literature. METHOD: A two and half year old girl presented to the University of Maiduguri Teaching Hospital. A pertinent clinical evaluation with abdominal ultrasound allowed accurate pre-operative diagnosis. RESULTS: A young girl with a right hypochondrial mass confirmed as choledochal cyst by ultrasound scans. Haematological parameters were normal but deranged clotting profiles were corrected with vitamin-K injection before surgical drainage. CONCLUSION: High index of suspicion with abdominal ultrasound scan allowed accurate pre-operative diagnosis of choledochal cyst that was successfully managed by surgical drainage, when attempted dissection of cyst wall for excision failed. PMID- 15532238 TI - Clinical photographic quiz. PMID- 15532240 TI - Health promoting behaviors among African American women with faith-based support. AB - The purpose of this qualitative exploratory study was to document health perceptions, beliefs and attitudes, intentions and social pressures that influence health promoting behaviors as expressed by community level aggregates of African American women with faith support. Twenty-six African American women from two large urban congregations with an active health ministry program participated in this study. Focus group interviews guided by the Theory of Planned Behavior (Ajzen & Fishbein, 1980 & 1991) were used to identify salient health perceptions, beliefs and attitudes, intentions, and social pressures influencing health-promoting behaviors in African American women with faith-based support. Positive health perceptions, attitudes, and beliefs were identified as important to engaging in a healthy lifestyle. Exercise, well balanced diets, weight reduction and stress management were the most salient health concerns among the respondents. Key referents identified included the pastor, congregational nurses, physicians, and church/family members. Control beliefs among these women reflected salient spiritual and fatalistic beliefs concerning health-promoting behaviors. Trusting relationships, open communication, safe, comfortable, and familiar environments were identified as important considerations when planning health promotion interventions for an African American faith community. CONCLUSIONS: Health beliefs, attitudes, and behaviors are not developed outside of social systems, therefore, the facilitation of healthy lifestyle behaviors may be best assessed and influenced within a context of reciprocal social interaction such as in a faith-based community. In the context of a community level aggregate with faith-based support, behaviors to promote a healthy lifestyle may be positively influenced. PMID- 15532241 TI - Kuwait: a personal cultural encounter after the storm. AB - The author discusses her experiences in traveling to Kuwait during the past several years and what she has learned about the people, their culture and the region. PMID- 15532242 TI - Transmission of HIV/AIDS among African American intravenous drug users: implication for public health policy. AB - According to recent research, Baltimore City has one of the highest HIV/AIDS (Human Immunodeficiency Virus), (Acquired Immunodeficiency Syndrome) infection rates in the state of Maryland. Intravenous drug use (IDU) in persons who are infected with HIV, is identified as the critical link to the spread of this disease in Baltimore. Substance abuse programs are insufficient to aid the city in helping to fight the warfare on HIV/ AIDS. The City Council, citizens and community based organizations must play a non-judgmental role in obtaining legislation to address the issues. Adequate substance abuse programs will greatly enhance educational and prevention strategies among African Americans who are already disproportionately affected by HIV/AIDS and IDU. Efficient, effective, equal, legal, political acceptance, and improvement would be included in the criteria of policy options. PMID- 15532243 TI - [Early initiation of statin therapy in patients with acute myocardial infarction after successful percutaneous coronary intervention]. AB - OBJECTIVES: To evaluate the effect of statins on the prognosis of acute myocardial infarction after percutaneous coronary intervention (PCI). METHODS: We reviewed 280 patients with acute myocardial infarction who underwent PCI within 12 hr after the onset of symptoms. Statin therapy was initiated in 72 patients within 8.6 +/- 7.6 days after the onset (statin group) but not in the remaining 208 (no statin group). The time sequential changes of low-density lipoprotein cholesterol (LDL-C) and C-reactive protein (CRP) levels, and the angiographic findings at 6 months after PCI were compared. RESULTS: At onset, LDL-C levels in the statin group were significantly higher than those in the no statin group (140 +/- 35 vs 118 +/- 28 mg/dl, p < 0.01). However, at restudy, the values were similar between the two groups (113 +/- 19 vs 118 +/- 21 mg/dl, p = 0.19). CRP levels at restudy tended to be lower in the statin group than in the no statin group (0.11 +/- 0.12 vs 0.14 +/- 0.13 mg/dl, p = 0.07). Although the binary restenosis rates of the culprit lesion were almost equivalent (statin group 29% vs no statin group 23%, p = 0.30), new lesions in the non-culprit vessels tended to be found more frequently in the no statin group than in the statin group (13% vs 4%, p = 0.07). CRP levels at restudy were significantly higher in the patients with new lesions (n = 27) than in those without (n = 253; 0.25 +/- 0.17 vs 0.11 +/- 0.19 mg/dl, p < 0.01), whereas LDL-C levels were similar between the two groups (117 +/- 20 vs 113 +/- 27 mg/dl, p = 0.75). LDL-C, CRP at restudy and the rates of new lesions were similar in the patients receiving water-soluble statins (n = 42) and liposoluble statins (n = 30). CONCLUSIONS: Statin therapy initiated at the early phase of acute myocardial infarction might prevent the development of new lesions in non-culprit vessels without any influence on the restenosis rate of the culprit lesion. PMID- 15532244 TI - [Relationship between redox state of whole arterial blood glutathione and left ventricular function after acute myocardial infarction]. AB - OBJECTIVES: To investigate the role of oxidative stress in left ventricular function after acute myocardial infarction. METHODS: We studied 41 patients with acute myocardial infarction (30 men and 11 women, mean age 61.7 +/- 11.6 years) with Thrombolysis in Myocardial Infarction grade 3 recanalization of occluded coronary arteries within 12 hr after onset. Cardiac catheterization was performed at the time of admission and before discharge. Three markers for oxidative stress were measured: plasma lipid hydroperoxide, plasma creatol and whole arterial blood glutathione at the time of admission. RESULTS: Mean time from onset to recanalization was 5.2 +/- 0.6 hr. The patients were divided into two groups according to the changes in left ventricular wall motion (LVWM); patients who showed improvement in LVWM and those without improvement. There were no significant differences in age, sex, coronary risk factors, severity of coronary artery disease, time from onset to recanalization or ejection fraction between two groups. Maximum creatine kinase and C-reactive protein levels in patients without LVWM improvement were significantly higher than in patients with improvement. Plasma levels of lipid hydroperoxide and creatol did not differ significantly between two groups. On the other hand, reduced glutathione/oxidized glutathione ratio in arterial blood in patients without LVWM improvement was significantly lower than in patients with LVWM improvement (69.8 +/- 3.4 vs 85.5 +/- 2.9, p < 0.05). CONCLUSIONS: Our results suggest that whole arterial blood glutathione is more oxidized in acute myocardial infarction patients without LVWM improvement than in patients with improvement. Redox state of arterial blood can be a predicting factor for left ventricular function after acute myocardial infarction. PMID- 15532245 TI - [Effect of hemodialysis on peak velocity of early diastolic mitral annulus motion]. AB - OBJECTIVES: Peak velocities of early diastolic mitral annulus motion (Ea) assessed by tissue Doppler imaging are relatively preload independent. Whether Ea decreases after hemodialysis or not is unclear. This study investigated the effect of hemodialysis on Ea. METHODS: The study group consisted of 23 hemodialysis patients without organic heart disease. Echocardiography was performed before and after hemodialysis, and Ea was compared with the peak velocity of early mitral inflow (E). Decreases in the body weight, E and Ea were calculated as indexes of decrease by hemodialysis. RESULTS: After hemodialysis, E decreased significantly (p = 0.005) but Ea did not change (p = 0.09). The decrease in Ea was significantly smaller than that in E (p < 0.0001). The decrease in E correlated significantly (r = 0.52, p < 0.05) with the decrease in the body weight, but the decrease in Ea did not correlate with the decrease in the body weight. CONCLUSIONS: Ea is little affected by hemodialysis compared with E. The effect of hemodialysis on Ea can be ignored if the amount of fluid removal is small. PMID- 15532246 TI - [Survivor of cardiogenic shock following acute myocardial infarction with Churg Strauss syndrome: first angiographic documention of coronary recanalization of infarct-related arteries: a case report]. AB - A 42-year-old man was treated under a diagnosis of Churg-Strauss syndrome with predonisolone pulse therapy. Three days later, he developed cardiogenic shock following acute myocardial infarction. Coronary angiography showed total occlusions in three peripheral coronary vessels. Intraaortic balloon pumping was used to maintain hemodynamics and predonisolone pulse therapy was repeated. However, he developed cardiogenic shock again after the second pulse therapy and needed percutaneous cardiopulmonary support and intraaortic balloon pumping. Accordingly, combination therapy of predonisolone and cyclophosphamide was given. He then recovered. Follow-up angiography showed recanalization of the infarct related arteries. PMID- 15532247 TI - [Inhibition of Rho-kinase by fasudil preventing anginal attacks associated with spastic angina: a case report]. AB - An 86-year-old woman was admitted with unstable angina pectoris. Plain old balloon angioplasty (POBA) was performed for 90% stenosis at segment 7 of the left coronary artery with concomitant treatment with nitrate, calcium antagonists, and nicorandil. Five days after POBA, she again suffered chest pain at rest with ST depression by electrocardiography, despite increased doses of calcium-antagonist and nicorandil. Coronary arteriography showed no evidence of restenosis (50%) at the POBA site. The involvement of coronary artery spasm was considered and intravenous treatment with a Rho-kinase inhibitor, fasudil, was started, which resulted in disappearance of the anginal attacks. She refused to continue the fasudil treatment on day 5, which resulted in reappearance of anginal attacks. Third coronary angiography showed a 90% restenosis at POBA site and percutaneous coronary intervention was again performed. This case suggests that a Rho-kinase inhibitor is potentially effective to prevent anginal attacks in spastic angina. PMID- 15532248 TI - [Incidental identification of cardiac disease based on high-pitched systolic murmur]. PMID- 15532249 TI - Working with African American clients: considering the "homeplace" in marriage and family therapy practices. AB - In this article, we discuss perspectives on the "homeplace" that are important to consider in marriage and family therapy involving African American clients. The homeplace comprises individual and family processes that are anchored in a defined physical space that elicits feelings of empowerment, rootedness, ownership, safety, and renewal. Critical elements of the homeplace include social relationships that shape individuals' and families' sense of social and cultural identity. We draw on our ethnographic and clinical research with African American families in urban and rural settings to describe typical schisms between therapists and African American clients when communicating about the homeplace. We also explore the impact of homeplace disruptions on experiences of "yearning." Recommendations for integrating a homeplace perspective into therapy practices are provided. PMID- 15532250 TI - Culturally sensitive adaptation of PREPARE with Japanese premarital couples. AB - In this study, we attempted to achieve a culturally sensitive adaptation of the PREmarital Personal And Relationship Evaluation (PREPARE) Inventory with Japanese premarital couples (n = 849). A translation, back-translation, and construct modification process was initiated by a group of U.S. and Japanese researchers to ensure a culturally sensitive and applicable adaptation of PREPARE. Alpha reliability analyses revealed good internal consistency among Japanese PREPARE scales. A Japanese premarital couple typology was developed using cluster analyses. Several unique features of the Japanese premarital types are identified and discussed. Suggestions are provided for continued enhancement in adaptation process of PREPARE with Japanese premarital couples. PMID- 15532252 TI - Predictors of attachment styles of children in foster care: an attachment theory model for working with families. AB - The purpose of this study was to examine the attachment styles of children in foster care and to use this information to inform clinical practice (N = 110). When examining the relationship between attachment style and child characteristics, no significant differences were found. A structural equation model hypothesizing the relation of individual characteristics, losses, and family contact to avoidant attachment was then tested. Results provided tentative support for the hypothesized model. Implications for therapists include being attentive to attachment injuries, and working to create safe therapeutic environments when working with families involved in the foster care system. PMID- 15532251 TI - The last mile of the way: understanding caregiving in African American families at the end-of-life. AB - This research is based on in-depth ethnographic interviews and focus groups with 88 African American family caregivers from various regions of the United States during a stressful time in their family development--caregiving at the end-of life--and the grieving during the aftermath. The study employed a stratified purposeful sampling strategy. Subjects were African Americans from the Northern, Southern, and Midwestern United States. Formal care is complicated by the distrust that many African Americans hold toward the health care system, which has resulted from years of exclusion, racism and discrimination. The findings highlight the importance of hearing from African American families to gain an understanding of what services, including family therapy and other psychotherapy, they will need during this process. PMID- 15532253 TI - Is Bowen theory valid? A review of basic research. AB - In the last 15 years, a substantial number of studies have tested the theoretical validity of Bowen family systems theory. A review of this basic research provided empirical support for the relationship between differentiation and chronic anxiety, marital satisfaction, and psychological distress. Bowen's assumption that couples with the same level of differentiation marry was not supported. His specific theories of sibling position and triangulation also received little empirical support. Research on multigenerational transmission has generally ignored Bowen's theoretical perspective, and more research needs to test Bowen's claim that his theory is universal. In addition, researchers still need to examine the effect of differentiation on child functioning, physical health problems, and adaptability. PMID- 15532254 TI - The core variables of symbolic-experiential therapy: a qualitative study. AB - Symbolic-experiential therapists have been criticized for not adequately operationalizing symbolic theory and empirically validating their methods. Although pioneering researchers have noted that the task of studying and evaluating humanistic therapies is complex, we agree that all therapists need to be held accountable for their interventions. In this article we identify the core variables underlying symbolic-experiential therapy that emerged from a research study using grounded theory. The inductively derived conceptual framework for symbolic-experiential therapy included: (a) Generating an interpersonal set; (b) creating a suprasystem; (c) stimulating a symbolic context; (d) activating stress within the system; (e) creating symbolic experience; and (f) moving out of the system. PMID- 15532255 TI - Training family therapists to work with children and families: a modified Delphi study. AB - This study examined child inclusion issues and training marriage and family therapists (MFTs) to treat children. This modified Delphi study utilized a panel of experts, and gathered data through questionnaires and qualitative interviews. Panelists believe children should participate in family therapy sessions for both child and adult problems, except when parents are discussing sex or sensitive issues. Child-focused courses should emphasize developmental issues, engaging techniques, theoretical issues, play therapy theory, MFT treatment for child disorders, and specific child/family problems. Panelists suggest numerous child focused references, but reached consensus for only one. Key therapist attributes and skills were identified. Deductive and inductive training methods and the role of supervision were highlighted. Although therapist playfulness and creativity were emphasized, few play techniques were included in the final profile. PMID- 15532256 TI - Cultural dissonance among generations: a solution-focused approach with East Asian elders and their families. AB - In traditional East Asian cultures, high value is assigned to family harmony and filial piety coupled with the expectation that elders will be honored and obeyed. A lifetime of such expectations shapes how elders perceive their role and status in the family. Problems can arise when younger, less traditional, generations do not share these expectations. This article describes a solution-focused approach that facilitates the family in creating a beneficial harmony in situations of cultural dissonance. Family members are empowered to draw on personal strengths in which multiple worldviews and values of individual members are recognized, incorporated, and negotiated. PMID- 15532257 TI - Estimating the cost of direct reimbursement of marriage and family therapy under Medicare. AB - In this article, we investigated the estimated cost to the Medicare program for covering psychotherapy services provided by marriage and family therapists (MFTs). Historical trends were identified by using psychotherapy cost and utilization data for the years 1999-2001. Using these trends, projections for the years 2002-2006 were made with MFTs included as providers. Employing this methodology, the 5-year estimated net increase and gross increase in cost due to the provision of psychotherapy services by MFTs was found to be approximately dollar 10.5 million (or dollar 2.1 million per year) and dollar 13.9 million (or dollar 2.8 million per year), respectively. This represents an increase of less than 1/2 of 1% of the Medicare mental health budget, and less than .0015% of Medicare expenditures overall. PMID- 15532258 TI - Static point-spread function correction dominating higher-order speckle terms at high adaptive correction. AB - At high adaptive correction, the randomly shifting speckles familiar in conventional astronomical imaging become organized into patterns with distinct regularities that may permit partial suppression of the image noise they produce. Mathematically, the phase exponential in the Fourier-optical imaging expression may be expanded in a Taylor series in remnant phase phi, which is small at very high correction, leading to a perturbed point-spread function (PSF) that is a sum of algebraic terms, each of distinct spatial symmetry. At sufficiently high correction, one need deal with only a few of the lowest-order terms. A first order expansion gives an ideal PSF plus two terms, linear and quadratic, describing the two brightest, physically most relevant kinds of speckle. A second order expansion gives three new terms, the brightest of which is primarily a static correction to the PSF, with a much smaller true speckle component. When the correction is great enough to isolate individual speckle terms, the two terms from the first-order expansion alone determine the essential physics. A general observational strategy is outlined for reducing speckle noise in highly corrected companion searches, dominated by a few speckle terms of definite spatial symmetry. PMID- 15532259 TI - Spatial and temporal coherence characterization of a smoothed laser beam. AB - The measurement of the coherence characteristics of the speckles generated by an optically smoothed laser source is investigated. We present a new method that can be used for every kind of smoothing technique. A modified Mach-Zehnder interferometer allows us to measure both the lifetime and the spatial transverse size of the hot spots generated by a broadband and transverse multimode source. Experimental results agree well with theoretical predictions. PMID- 15532260 TI - Measurement of gouy phase evolution by use of spatial mode interference. AB - An experimental technique to observe and accurately measure the Gouy phase evolution of Hermite-Gaussian modes is presented. Because of the unique features of spatial mode interference frequency-locking error signals, we are able to readily perform explicit measurement of the Gouy phase in a simple and highly accurate manner. We present these data and discuss the technique and its implications. PMID- 15532261 TI - Low-loss grating for coupling to a high-finesse cavity. AB - A concept for a low-loss all-reflective cavity coupler is experimentally demonstrated at a wavelength of 1064 nm. A 1450-nm period dielectric reflection grating with a diffraction efficiency of 0.58% in the - 1st order is used in the 2nd-order Littrow configuration as a coupler to form a cavity with a finesse of 400. The application of such reflective low-loss cavity couplers in future generations of gravitational-wave detectors and implementation issues are discussed. PMID- 15532262 TI - Improved first Rayleigh-sommerfeld method for analysis of cylindrical microlenses with small f-numbers. AB - An improved first Rayleigh-Sommerfeld method (IRSM1) is proposed and applied to the analysis of cylindrical microlenses with small f-numbers. Numerical results obtained by both the IRSM1 and the original Rayleigh-Sommerfeld method (ORSM1) are compared with those obtained by the rigorous boundary element method (BEM). For both refractive and diffractive lenses, the results obtained by the IRSM1 are close to those obtained by the BEM even for small f-numbers; by contrast, the results by the ORSM1 differ significantly from those obtained by the BEM. Moreover, the IRSM1 uses much less time and computer memory in the computations than the BEM. PMID- 15532263 TI - Mixed data rate and format transmission (40-Gbit/s non-return-to-zero, 40-Gbit/s duobinary, and 10-Gbit/s non-return-to-zero) by mid-link spectral inversion. AB - A polarization-diverse subsystem based on periodically poled lithium niobate waveguides is used as an optical phase conjugator for compensation for linear and nonlinear distortion. We show successful transmission formats of 13 x 40 Gbit/s non-return-to-zero mixed with 6 x 10 Gbit/s non-return-to-zero and 40-Gbit/s duobinary over 8 x 100 km of standard single-mode fiber. A single phase conjugator is used to conjugate all data formats, including the alternative duobinary format, simultaneously. PMID- 15532264 TI - Wideband frequency modulation of a mode-locked fiber laser. AB - We have demonstrated wideband frequency modulation of the frequency comb lines of a high-repetition-rate fiber laser. With a modulation frequency of only approximately 10 kHz, we have generated modulation indices in excess of 250. Although internally modulated, the laser remains stable with 2-kHz linewidths, and thus the 10-kHz modulation sidebands are still clearly resolved even after propagation over several hundred kilometers. This unique characteristic is used for simultaneous measurement of propagation distances to 1-m resolution and velocities of less than 3 mm/s over a distance of greater than 50 km. PMID- 15532265 TI - Reduction of collision-induced timing shifts in dispersion-managed quasi-linear systems with periodic-group-delay dispersion compensation. AB - Periodic-group-delay (PGD) dispersion-compensation modules were recently proposed as mechanisms to alleviate collision-induced timing shifts in dispersion-managed (DM) systems. Frequency and timing shifts in quasi-linear DM systems with PGDs were obtained, and it is shown that significant reductions are achieved when even a small fraction of the total dispersion is compensated for by PGDs. PMID- 15532266 TI - All-optical regeneration of differential phase-shift keying signals based on phase-sensitive amplification. AB - All-optical regeneration of differential phase-shift keying signals based on phase-sensitive amplification is described. Nearly ideal phase regeneration can be achieved in the undepleted-pump regime, and simultaneous amplitude and phase regeneration can be realized in the depleted-pump regime. PMID- 15532267 TI - Enhanced type IIA gratings for high-temperature operation. AB - The inscription of type IIA fiber Bragg gratings in standard boron-codoped germanosilicate fiber has been demonstrated to show marked differences from that reported in the literature. These gratings were subjected to high temperatures, and their decay behavior was evaluated. Gratings resistant to heat up to 800 degrees C for a moderate length of time are demonstrated. PMID- 15532268 TI - Supercontinuum generation in ultraviolet-irradiated fibers. AB - We demonstrate that UV exposure of highly nonlinear, germanosilicate fibers causes a strong change in their chromatic dispersion and can significantly alter the infrared supercontinuum generation in these fibers. By varying the level of UV exposure to the fiber, we show that the dispersion zero and the short wavelength edge of the supercontinuum can be changed by more than 100 nm. A nonlinear Schrodinger equation model of the continuum generation in the nonlinear fiber shows that the short-wavelength behavior of the continuum is primarily controlled by changes in the fiber dispersion caused by the UV-induced change in the refractive index of the fiber core. PMID- 15532269 TI - Nonparaxial free-space diffraction from oblique end faces of single-mode optical fibers. AB - We investigate free-space diffraction of light that emanates from obliquely cleaved end faces of single-mode fibers. Emphasis is placed on precise prediction of the wave-front sphericity of fiber-generating waves in the nonparaxial Fresnel propagation region spanning an entire hemispherical observation surface. Rayleigh Sommerfeld scalar diffraction theory is used to produce an analytic closed-form solution with a nonparaxial approximation. The result allows the wave front's sphericity and the amplitude distribution of fiber-generating waves to be evaluated precisely with less computation than for existing numerical or infinite series solutions. PMID- 15532270 TI - All-solid photonic bandgap fiber. AB - We describe the design and fabrication of a photonic bandgap fiber formed with two different glasses. As in a hollow-core fiber, light is guided in a low-index core region because of the antiresonances of the high-index strands in the fiber cladding. The structure described represents an ideal bandgap fiber that exhibits no interface modes and guides over the full width of multiple bandgaps. PMID- 15532271 TI - Noise-figure limit of fiber-optical parametric amplifiers and wavelength converters: experimental investigation. AB - Recent theoretical work predicts that the quantum-limited noise figure of a x(3) based fiber-optical parametric amplifier operating as a phase-insensitive in-line amplifier or as a wavelength converter exceeds the standard 3-dB limit at high gain. The degradation of the noise figure is caused by the excess noise added by the unavoidable Raman gain and loss occurring at the signal and the converted wavelengths. We present detailed experimental evidence in support of this theory through measurements of the gain and noise-figure spectra for phase-insensitive parametric amplification and wavelength conversion in a continuous-wave amplifier made from 4.4 km of dispersion-shifted fiber. The theory is also extended to include the effect of distributed linear loss on the noise figure of such a long length parametric amplifier and wavelength converter. PMID- 15532272 TI - Three-dimensional polarimetric integral imaging. AB - A three-dimensional (3D) polarimetric image sensing and display technique based on integral imaging is proposed. Three-dimensional polarization distribution of reflected light from a 3D object can be measured as elemental image arrays by a rotating linear polarizer. After the measurement of the polarization of the 3D object, the 3D polarimetric object can be reconstructed optically by displaying the polarization-selected elemental images in spatial light modulators with two quarter-wave plates. Experimental demonstration of 3D polarimetric imaging of a 3D object attached to two orthogonal linear polarizers is presented. To the best of our knowledge, this is the first report on 3D polarimetric sensing imaging and 3D optical reconstruction by integral imaging. PMID- 15532274 TI - Application of an optical pulse stretcher to coherent anti-Stokes Raman spectroscopy. AB - An external optical cavity pulse stretcher for nanosecond-long laser pulses has been applied to coherent anti-Stokes Raman spectroscopy (CARS). An increased signal-to-noise ratio was achieved for both vibrational and pure rotational CARS, while the power density of the laser beams remained constant. Moreover, it was demonstrated that the use of the pulse stretcher also leads to improved precision of the determined temperatures and concentrations as a result of repeated excitation of the dye laser. PMID- 15532273 TI - Highly sensitive differential phase-sensitive surface plasmon resonance biosensor based on the Mach-Zehnder configuration. AB - A high-sensitivity surface plasmon resonance (SPR) biosensor based on the Mach Zehnder interferometer design is presented. The novel feature of the new design is the use of a Wollaston prism through which the phase quantities of the p and s polarizations are interrogated simultaneously. Since SPR affects only the p polarization, the signal due to the s polarization can be used as the reference. Consequently, the differential phase between the two polarizations allows us to eliminate all common-path phase noise while keeping the phase change caused by the SPR effect. Experimental results obtained from glycerin-water mixtures indicate that the sensitivity limit of our scheme is 5.5 x 10(-8) refractive index units per 0.01 degrees phase change. To our knowledge, this is a significant improvement over previously obtained results when gold was used as the sensor surface. Such an improvement in the sensitivity limit should allow SPR biosensors to become a possible replacement for conventional biosensing techniques based on fluorescence. Monitoring of the bovine serum albumin (BSA) binding reaction with BSA antibodies is also demonstrated. PMID- 15532275 TI - Eliminating the birefringence in silicon-on-insulator ridge waveguides by use of cladding stress. AB - We propose and demonstrate the use of the cladding stress-induced photoelastic effect to eliminate modal birefringence in silicon-on-insulator (SOI) ridge waveguides. Birefringence-free operation was achieved for waveguides with otherwise large birefringence by use of properly chosen thickness and stress of the upper cladding layer. With the stress levels typically found in cladding materials such as SiO2, the birefringence modification range can be as large as 10(-3). In arrayed waveguide grating demultiplexers that were fabricated in a SOI platform, we demonstrated the reduction of the birefringence from 1.2 x 10(-3) (without the upper cladding) to 4.5 x 10(-5) when a 0.8-microm oxide upper cladding with a stress of -320 MPa (compressive) was used. Because the index changes induced by the stress are orders of magnitude smaller than the waveguide core-cladding index contrast, the associated mode mismatch loss is negligible. PMID- 15532276 TI - Optical bistability on a silicon chip. AB - We demonstrate, for the first time to our knowledge, optical bistability on a highly integrated silicon device, using a 5-microm-radius ring resonator. The strong light-confinement nature of the resonator induces nonlinear optical response with low pump power. We show that the optical bistability allows all optical functionalities, such as switching and memory with microsecond time response and a modulation depth of 10 dB, driven by pump power as low as 45 microW. Silicon optical bistability relies on a fast thermal nonlinear optical effect presenting a 500-kHz modulation bandwidth. PMID- 15532277 TI - Efficient continuous-wave generation in a self-organizing diode-pumped Nd:YVO4 laser with a reciprocal dynamic holographic cavity. AB - A diode-pumped Nd:YVO4 laser is demonstrated that has a reciprocal self intersecting loop cavity with self-organization by formation of a dynamic holographic grating induced by the generating beam. Continuous-wave high spatial quality generation (M2 < 1.2) with 9.6 W of power and 32% optical efficiency is obtained. Spectral compositions and temporal dynamics are investigated as functions of diode-pumping power. PMID- 15532278 TI - Experimental synchronization of chaos in diode lasers with polarization-rotated feedback and injection. AB - We demonstrate experimental chaos synchronization between two chaotic semiconductor lasers subjected to polarization-rotated optical feedback and unidirectional injection. This system allows high-quality synchronization to be obtained between dissimilar lasers in a wide range of chaotic operating regimes. Another feature of this system is its operation at high characteristic frequencies, taking advantage of all-optical implementation. Time series and RF spectra showing synchronization are confirmed by high correlation coefficients in excess of 0.85. PMID- 15532279 TI - Efficient single-axial-mode operation of a Ho:YAG ring laser pumped by a Tm-doped silica fiber laser. AB - Efficient single-frequency operation of a Ho:YAG ring laser at room temperature with a traveling-wave TeO2 acousto-optic modulator to enforce unidirectional operation is reported. By use of a 2-at. % Ho3+-doped 10-mm-long Ho:YAG rod, end pumped by a cladding-pumped tunable Tm-doped silica fiber operating at 1.9 microm, the Ho:YAG ring laser yielded 3.7 W of single-frequency output at 2.1 microm in a diffraction-limited TEM00 beam with M2 < 1.1 for an incident pump power of 8.8 W. The rf power required for unidirectional operation was 0.3 W and corresponded to an increase in cavity loss for the lasing direction (due to diffraction) of only 0.5%. The prospects for further improvement in efficiency are discussed. PMID- 15532280 TI - Quantitative phase imaging using actively stabilized phase-shifting low-coherence interferometry. AB - We describe a quantitative phase-imaging interferometer in which phase shifting and noise cancellation are performed by an active feedback loop using a reference laser. Depth gating via low-coherence light allows phase measurement from weakly reflecting biological samples. We demonstrate phase images from a test structure and living cells. PMID- 15532281 TI - Determination of local polarization properties of biological samples in the presence of diattenuation by use of Mueller optical coherence tomography. AB - A unique feature of polarization-sensitive Mueller optical coherence tomography is that, by measuring Jones or Mueller matrices, it can reveal the complete polarization properties of biological samples, even in the presence of diattenuation. We map local polarization properties for the first time to our knowledge by using polar decomposition in combination with least-squares fitting to differentiate measured integrated Jones matrices with respect to depth. We also introduce the new concept of dual attenuation coefficients to characterize diattenuation per unit infinitesimal length in tissues. We experimentally verify the algorithm using measurements of a section of porcine tendon and the septum of a rat heart. PMID- 15532283 TI - Four-wave mixing of linear waves and solitons in fibers with higher-order dispersion. AB - We derive phase-matching conditions for four-wave mixing between solitons and linear waves in optical fibers with arbitrary dispersion and demonstrate resonant excitation of new spectral components via this process. PMID- 15532282 TI - Double-clad fiber for endoscopy. AB - Endoscopes employing a single optical fiber may have advantages over conventional fiber-bundle or CCD array imaging techniques, including the potential for greater flexibility and miniaturization. Although single-mode fibers can provide superior resolution compared with multimode fibers, they are prone to increased speckle noise and suffer from limited optical throughput and reduced depth of field. We demonstrate the use of a double-clad fiber for single-mode illumination and multimode detection to achieve high-resolution, reduced-speckle imaging with high optical throughput and a large depth of field. PMID- 15532284 TI - Nonlinear refractive index of a rare-earth-doped fiber laser. AB - An expression is derived for the intensity-dependent index of refraction of a rare-earth-doped fiber laser by use of a Kramers-Kronig relation and simple rate equations. The solution is applied to examine the properties of the nonlinear index of an ytterbium-doped silicate fiber laser at 975-nm pump and 1080-nm laser wavelengths. PMID- 15532286 TI - Wide-viewing projection-type integral imaging system with an embossed screen. AB - A projection-type integral imaging system with an embossed screen is proposed to realize a wide viewing angle for the reconstructed image. The embossed screen is designed for virtual integral imaging in a projection-type system. It can enhance the viewing angle by increasing the effective region for each elemental image. The experimental result that verifies the merit of the proposed method is presented. PMID- 15532285 TI - Measurement of polarization dependence of nonlinear susceptibility responsible for Rayleigh-wing and Brillouin scattering. AB - By using a simple optical geometry based on backward light scattering and employing a Sandercock-type tandem Fabry-Perot interferometer, we measure both the linear and the circular polarization dependences of Rayleigh-wing and Brillouin scattering in a sample of liquid-crystal 4-n-pentyl-4'-cyanobiphenyl. Observed polarization dependences are consistent with the third-order nonlinear susceptibilities, taking into account the traceless symmetric scattering tensor for Rayleigh-wing scattering and the isotropic scattering tensor for Brillouin scattering. PMID- 15532287 TI - Detection of hidden stationary deformations of vibrating surfaces by use of time averaged digital holographic interferometry. AB - A method of detecting displacements of a surface from its steady-state position to its equilibrium position while it is vibrating has been developed by use of time-average digital holographic interferometry. This method permits extraction of such a hidden deformation by creating two separated systems of interferogram fringes: one corresponding to a time-varying resonantly oscillating optical phase, the other to the stationary phase modification. A mathematical description of the method and illustrative results of experimental verification are presented. PMID- 15532288 TI - All-fiber narrowband polarization controller based on coherent acousto-optic mode coupling in single-mode fiber. AB - An all-fiber, narrowband, tunable polarization controller is proposed and demonstrated. The device is based on coherent acousto-optic mode coupling induced by two orthogonal acoustic waves on a dispersion-compensating fiber. The cooperative coupling between the two polarizations of the core mode and the TE01 cladding mode through the two gratings permits indirect coupling between the two polarizations of the core mode with nearly 100% efficiency, which makes the polarization-controlling function possible. Experimental results verify the operation of the polarization controller with an insertion loss of <1 dB. PMID- 15532289 TI - Enhancing the axial resolution of quantum optical coherence tomography by chirped quasi-phase matching. AB - We show theoretically that a chirped quasi-phase-matching nonlinear crystal structure can significantly enhance the axial resolution in quantum optical coherence tomography by increasing the spectral width of the generated entangled photon pairs. For pulsed pumps we show how the pump-pulse duration affects the maximum resolution attainable. PMID- 15532290 TI - Temporal compression of short-wavelength laser pulses by coherent control in rare gases. AB - We present a theoretical study of temporal compression of a short-wavelength laser pulse predicted in a real, Doppler-broadened, atomic system. The compression is the result of the coherent control peculiarities of electromagnetically induced transparency-propagation dynamics. Numerical results are reported and discussed, showing a temporal compression of 2 orders of magnitude (from 10 ns to 100 ps) of a 106.7-nm laser pulse in argon atoms at room temperature. PMID- 15532291 TI - Determining thickness independently from optical constants by use of ultrafast light. AB - We show that the application of ultrafast techniques, especially terahertz time domain spectroscopy, allows simultaneous measurements of material thickness and optical constants from transmission measurements, by analyzing not only the phase difference between the main terahertz pulse through the medium but also the subsequent multireflection pulse (an echo) from the medium. Such a method provides a fast and precise characterization of the optical properties and can extract thickness information and hence other optical constants in a broad bandwidth. It may have applications in science and engineering such as in situ film thickness and quality monitoring, optical constants measurement, medical imaging, noninvasive detection, and remote sensing. PMID- 15532293 TI - The spread of value-based purchasing depends on the corporate employer. PMID- 15532292 TI - Aligning incentives in bridges to excellence. PMID- 15532294 TI - Can incentives be aligned successfully? PMID- 15532295 TI - The need for collaboration. PMID- 15532297 TI - Does value-based purchasing mean smaller provider networks? PMID- 15532296 TI - Motivating the employee to participate. PMID- 15532298 TI - Does ischemic preconditioning occur during rehabilitation of ischemic patients? AB - Ischemic preconditioning is generally a short duration phenomenon, but during acute myocardial infarction a better inhospital outcome of patients with previous angina suggests that the effect of ischemic preconditioning could be longer than it was considered until now. It was also demonstrated that during successive exercise stress testings (ET) ST depression became lesser as a consequence of ischemic preconditioning. We investigated the possible occurrence of ischemic preconditioning during rehabilitation of ischemic patients. There were studied 43p with old myocardial infarction or stable effort angina, 29 men and 14 women, aged 33-68 years, divided in two groups: group I (19p) was included in a cardiac rehabilitation programme (15-20 sessions) and group II (24p) was not. All the patients were submitted to two ET at a month interval, for group I before and after cardiac rehabilitation programme. The peak-effort level was significantly greater for group I (80.26 +/- 7.2w vs 93.42 +/- 8.3w p < 0.05) but not for group II (65.5 +/- 5.8w vs 72.91 +/- 6.5w p > 0.05) during the two ET. The double product (21.573 +/- 3122 vs 24.168 +/- 3423 for group I and 23.551 +/- 3100 vs 21.000 +/- 2752 for group II) was not significantly different (p > 0.05) during the two ET. The same unsignificant modification was noted for maximal ST depression for group II (1.458 +/- 0.32 vs 1.166 +/- 0.21 mm p > 0.05) but not for group I patients in which maximal ST depression during the second ET was significantly less than during the first ET (1.52 +/- 0.23 vs 0.736 +/- 0.12 mm p < 0.05). Because cardiac rehabilitation does not influence the ischemic threshold and because the lesser maximal ST depression during the second ET was noted at the same, or even greater, double product (MVO2), we consider that it can be attributed to ischemic preconditioning. PMID- 15532299 TI - Endomyocardial biopsy in diagnosis of myocardial diseases. AB - In the past years, diagnosis of myocardial diseases became less difficult. A major contribution in this direction has the recent development of diagnosis methods. One of the most recent diagnosis procedures is the endomyocardial biopsy, which manages directly through the histologic examination to establish a clear diagnosis in myocardial diseases. This method provides faithful morphological data about the pathological changes in the myocardium. The aim of our study was to underline the importance of endomyocardial biopsy in an accurate diagnosis of different cardiomyopathies. In the period of 12 years (1986-1998), in the Cardiovascular Department of the First Medical Clinic Targu-Mures, Romania, were admitted 137 patients suffering from different cardiomyopathies. We performed a number of more than 100 endomyocardial biopsies on 25 of them, obtaining in each case 3 to 5 samples of endomyocardial tissue from the right ventricle. In all cases the Cordis bioptome was introduced in the right ventricle via femoralis vein. The complications were minor: ventricular extrasystoles. In all cases, we succeeded to obtain enough tissue for the histopathological examination, from different areas of RV. Histologic examination showed in all cases characteristic aspects of different cardiomyopathies. helping to clarify the diagnosis. After performing Endomyocardial Biopsy, we record an important increase of dilated cardiomyopathy percentage of diagnosis, from 40% to 72%. In patients with dilated cardiomyopathy, we observed a good correlation between hemodynamical data (Ejection Fraction, Left Ventricle End-Diastolic Pressure, etc.) and the index of myocardial damage, especially the degree of interstitial fibrosis. Using this method it is possible to distinguish different types of cardiomyopathies, such as: primary dilated cardiomyopathy, eosinophilic cardiomyopathy, myocarditis, idiopathic hypertrophic cardiomyopathy. Because the complications of this procedure are insignificant, and because of its important benefit for an accurate diagnosis, we recommend to apply this method for different cardiomyopathies. PMID- 15532300 TI - Immunological methods in the diagnosis of monoclonal gammopathies. The prognostic impact of the immunological data. AB - Immunological data are extremely important for the diagnosis of monoclonal gammopathies (MG). During the 1977-1998 period, 160 MG cases were diagnosed in our laboratory. Out of the 130 multiple myeloma (MM) cases, 70 (53.85%) met the strictly immunological criteria; in the rest of the cases, the immunological data had to be corroborated with the bone marrow cytology and bone X-ray data. We tried to assess retrospectively the prognostic impact of the immunological data on the 60 cases that were treated and followed at the Hematology Dept. of the Cancer Institute, Cluj. The following parameters were found to have a favorable prognostic impact on survival: IgG monoclonal component, kappa light chains, response to therapy with plateau phase achievement, Salmon Durie stages I and II at diagnosis. PMID- 15532301 TI - Hematopoietic stem cell transplantation in malignant lymphoproliferative diseases. A single center, retrospective study. AB - During the 1982-1997 period, out of the 133 hematopoietic stem cell (HSC) transplantations performed at Mont Godinne for malignant hemopathies, 85 were done in lymphoproliferative diseases: 27 in aggressive non-Hodgkin's lymphomas (ANHL), 21 in multiple myeloma (MM) patients, 17 in low-grade non-Hodgkin's lymphomas (LNHL), 9 in Hodgkin's disease (HD) cases, 9 in acute lymphoblastic leukemia (ALL) and 2 in chronic lymphocytic leukemia (CLL) patients. According to the HSC source, there were 19 allogeneic bone marrow grafts and 66 autologous HSC grafts (14 bone marrow and 52 peripheral stem cell grafts). In 40 cases the procedure was done after achieving partial or complete remission through conventional chemotherapy and in 45 patients in case of relapse or progressive disease. The conditioning regimens consisted of high-dose chemotherapy, associated with total body irradiation in case of allografts and in autografts for ALL. Engraftment was obtained in all autografted patients, transplant related mortality (TRM) occurring in 2 patients (3%). In the allogeneic transplantation group, 1 patient did not engraft and TRM was much higher, occurring in 10 cases (52%). Hematologic recovery occurred significantly faster in the autograft group than in the allograft group. The overall costs of an autograft were much lower than those of an allograft. Out of the 59 patients followed 4 years or longer, 23 (39%) are alive, free of disease, the proportion varying from 57% in the HD cases to 16% in the MM cases. The overall survival at 4 years was 49%. Negative prognostic factors for disease free survival at 4 years included male sex, lack of complete remission status at transplantation and MM diagnosis, whereas male sex and allografting were the negative prognostic factors for the 4-year overall survival. PMID- 15532302 TI - The levels of bilirubin may be related to an inflammatory condition in patients with coronary heart disease. AB - In agreement with previous reports, we found that the bilirubin level is significantly lower in the blood of patients with coronary heart disease (CHD) than in age and sex matched controls. However, we found that the level of bilirubin in the blood seemed to be an age-dependent phenomenon and closely related to the activation of leukocytes. In 1,000 cardiac catheterized patients from Urbana. USA suffering from CHD, the level of blood bilirubin was found to be lower than in age and sex-matched controls. The same results were obtained on 300 patients with acute ischemia from three hospitals from Bucharest, Romania. The activation of polymorphonuclear leukocytes increased in the catheterized patients, as well in Romanian patients. An activation of leukocytes triggered by a chronic inflammatory process may increase the lysis of erythrocytes. The erythrocytes of patients with 100% stenosis exhibited a higher rate of in vitro lysis in the presence of activated leukocytes and homocysteine. The increased hemolysis may trigger the activation and removal of the resulting bilirubin from blood. Such a mechanism may depend on the liver clearing function. This function had decreased in catheterized patients over 60 years of age, but had accelerated in younger patients. An individual variation in liver function may explain the widespread bilirubin levels in the blood of patients suffering from CHD. PMID- 15532303 TI - Disturbances of the coagulating equilibrium of blood in diabetes mellitus. AB - The diabetes mellitus is accompanied in its evolution by ample disturbances of the blood's fluid-coagulating equilibrium, which contribute to the initiation and development of the micro and macro-vascular complications. The early and accelerated atherosclerosis in diabetes mellitus obliged the medical world to the necessity to solve the pathogenic implied mechanisms: knowing them allowed us to apply a therapy that can prevent or improve the associated vascular pathology. The intense research made in the last decades mentioned that, in diabetes mellitus, all the stages of the hemostasis are affected, noticing the diminution of the fibrinolysis. The growth of the production of pro-aggregating factors unleashed from the platelets or other cells--TXA2. PAF, and the lowering of the synthesis of the protector endothelial factors--PGI2, NO--are followed by the intensifying adhesion and aggregation growth of the platelets at the injured endothelium, with the worsening of the atherosclerosis injuries. Furthermore, the activated platelets are keeping and are augmenting other steps of the hemostasis- first of all the coagulation, in this way contributing to the stage of hypercoagulability in diabetes mellitus. The deficit of plasminogen activators and the growth of the synthesis of fibrinolysis inhibitors also keep the stage of hypercoagulability, underlined by the growth of plasmatic concentration of the fibrinogen, especially PAI-1. There still are controversies concerning the stage of hypercoagulability whether it is primary in diabetes or it is an answer reaction to the endothelial damages and the high level of glycaemia. The role of the coagulation-fibrinolyses-thrombocytus-prostaglandins-CFTP in the initiating and development of the micro and macro-vascular complications, as well as in the acceleration of the atheriosclerosis process in diabetes mellitus, is nowadays certitude. More difficult is the attempt to identify the pathogenic mechanisms in which the CFTP sector interferes: the research in this domain still begins under the sign of incertitude. PMID- 15532304 TI - Has this patient a cancer? The assessment of weight loss, anemia and erythrocyte sedimentation rate as diagnostic tests in cancer. A retrospective study based in a secondary care university hospital in Romania. AB - INTRODUCTION: "Has this patient a cancer?" is a question we frequently ask ourselves in front of some symptoms, signs or laboratory results like weight loss, anemia and high erythrocyte sedimentation rate (ESR). The aim of this study is to assess their value in the diagnosis of cancer. METHODS: A retrospective study of the records of 5500 patients admitted in the Department of Internal Medicine during the 1998 January-September period has been performed, selecting those patients with weight loss, anemia (Hb < 10.5g/dl) and ESR > = 50 mm/h. RESULTS: The three diagnostic tests (anemia, high ESR and weight loss) had a low sensitivity (37, 52 and 46%, respectively), but a good specificity (92, 89 and 94%, respectively). For a 4% prevalence of cancer, as seen in our group, the resulting negative predictive value for the three tests was estimated respectively at 97, 98 and 98%, the post-test probabilities being estimated at 3, 2.2% in the case of negative tests. For a parallel utilisation of the tests, the sensitivity was 87%, the specificity 79%, the positive predictive value 15% and the negative predictive value 99%, the post-test probability in the case of a negative test being 1% [LR(-) = 0.17]; for serial utilisation, the specificity grew at 99.6% (CI: 99-100%), the positive predictive value (as the post-test probability in the case of a positive test) being 51% (CI: 50-53%) [LR(+) = 24.62]. INTERPRETATION: Any patient admitted in our department of Internal medicine has a 4% probability of having a cancer. Among those with weight loss, anemia and high ESR, one patient out of two has a cancer; among those with weight loss and anemia, 44% have a cancer; and among those with weight loss and high ESR, one out of three has a cancer. PMID- 15532305 TI - Silicosis related changes in serum copper. AB - To ascertain the diagnostic and prognostic value of serum copper in silicosis, this trace element was determined in 68 patients (all males), aged varied between 20 and 60 years (mean age 44 +/- 2.3 years). The same determination was carried out in a control group of 35 healthy subjects. The patients were distributed into three subgroups according to the evolutive stage of disease. The mean value of serum copper increased gradually from stage to stage reaching in stage III the highest copper mean value (p < 0.01). Serum copper concentrations also increased with the severity of the disease. These changes suggest that copper is directly responsible for the lung changes in silicosis and can be used as an indicator of the severity of the inflammatory reaction. PMID- 15532306 TI - Restless legs syndrome--relevant aspects for internal medicine specialists. AB - Restless legs syndrome (RLS), also known as Ekbom's syndrome, is a fairly common complaint which is not widely recognised by medical professionals, although it seems to affect 1-10% of the population. Despite recent attempts to better characterize RLS, this neurologic disorder remains poorly understood. Idiopathic RLS frequently follows an autosomal dominant inheritance with a variable clinical expressivity of symptoms. Secondary RLS is usually associated with neuropathy of chronic disorders (uremia, cryoglobulinemia, diabetes mellitus, infections, etc). RLS gives the sufferer an unpleasant sensation in the legs at rest, causing an irresistible desire to move which alleviates the discomfort. Other features that characterize RLS include sleep disturbance, involuntary movements in sleep or wakefulness, a normal neurologic examination, a chronic clinical course (waxing and waning over the time), and, in some cases, a positive family history. Periodic limb movements during sleep, which also may occur as an isolated finding, may or may not cause frequent arousals or awakenings. Clinical diagnosis of idiopathic or symptomatic forms of RLS can be supported with polysomnography. Full understanding of the features of RLS will provide the clinician with the strongest tool for recognizing the disorder. Many different treatments have been tried for RLS. Since the cause is unclear, therapy of RLS and PLMS remains symptomatic except for some secondary forms. Treatment of first choice consists of dopaminergic drugs or dopamine agonist, opioids and benzodiazepines. PMID- 15532307 TI - Assessment of treatment with orotate magnesium in early postoperative period of patients with cardiac insufficiency and coronary artery by-pass grafts (ATOMIC). AB - The benefit of the treatment with magnesium orotate (magnerot) was assessed in a randomised, single blind and placebo controlled study. Respecting the inclusion criteria were selected 32 patients with ischemia chronic failure in early postoperative period after CABG. The main improvements induced by magnesium orotate are the increase in exercise capacity (distance ambulated during 6 minutes walk test and ergospirometric parameters) and the reduction of ventricular premature beats. The treatment was well tolerated and the adverse reactions were not significant. The study strongly suggests the benefit of magnesium orotate added to classical antiischemic therapy in the complex management of coronary patients after CABG. PMID- 15532308 TI - Effects of 3-month treatment with the antioxidant alpha-lipoic acid in diabetic peripheral neuropathy. AB - Alpha lipoic acid is a natural antioxidant that has been suggested to improve symptoms of diabetic neuropathy. To assess these potential benefits, a cohort of 26 type 2 diabetic patients with symptomatic peripheral neuropathy (stage 2) received a daily dose of 600 mg alpha-lipoic acid, and were followed for a 3 month-period. No dropout was noted. At the end of the follow-up period, in 20 subjects (76.9%) there was a 1-stage regression of somatic neuropathy (from symptomatic to asymptomatic neuropathy, and in 5 patients (19.2%) no signs of neuropathy were found. The nerve conduction velocity of motor fibers improved from 36.8 (95% Confidence Interval CI = 30.9-42.7) meters/second to 41.3 (95% CI = 39.5-43.0) meters/second (p = 0.049, paired t test). Mean blood glucose measured was significantly lower at 3 months than at baseline [197.9 (95% CI = 170.1-225.7) versus 162.2 (95% CI = 146.1-178.2 mg/dl, p = 0.02, paired t test)]. In a multiple linear regression model with age. sex, body mass index, diabetes duration and the difference between blood glucose values at 3 months and at baseline as explanatory variables, the increment in nerve conduction velocity was not accounted for by the improved glycemic control. Women, thinner and younger patients tended to benefit more from the treatment in terms of nervous conduction velocity improvement. In conclusion, alpha-lipoic acid seems to be efficient and safe in the treatment of diabetic peripheral neuropathy, improving both clinical manifestations and nerve conduction velocity. Placebo controlled clinical trials are needed to further define the role of this new medication in the treatment of diabetic neuropathies. PMID- 15532309 TI - Following the money at the NIDCR. PMID- 15532310 TI - Affecting upper extremity strength by changing maxillo-mandibular vertical dimension in deep bite subjects. AB - The effect of vertical dimension of occlusion (VDO) on maximizing isometric deltoid strength (IDS) was measured in subjects with deep overbite. Sixteen female dental students with deep dental overbite and no history of temporomandibular joint disorder (TMD) were used as their own control and tested for isometric strength of the deltoid muscles, using a hand held strain gauge. Measurements were taken under four mandibular conditions: 1. habitual occlusion; 2. mandibular rest position; 3. biting on a bite elevating appliance set to the functional criterion of peak IDS; and 4. biting on a placebo appliance. Results showed that in deep bite subjects, isometric deltoid strength in habitual occlusion was significantly less than in the mandibular rest position. Isometric deltoid strength with the bite elevating appliance was significantly greater than isometric deltoid strength in habitual occlusion, as well as in the mandibular rest position. Isometric deltoid strength achieved in habitual occlusion and placebo did not differ. Results of this study support previous findings indicating that a change in the VDO will affect isometric strength of the upper extremities. PMID- 15532311 TI - Muscle hardness characteristics of the masseter muscle after repetitive muscle activation: comparison to the biceps brachii muscle. AB - The purpose of this study was to compare hardness characteristics of the masseter muscle to those of the biceps brachii muscle during repetitive muscle movements. Seventeen asymptomatic female subjects participated in this study. Each subject, on separate days, undertook a 5-minute unilateral chewing gum task on the right side and a 5-minute flexion-extension exercise on the right hand with a 2kg dumbbell. Using a handheld hardness meter, muscle hardness was measured in the right masseter and in the biceps brachii muscle at eight time points (before the task, immediately after the task, and at 1, 3, 5, 10, 30, and 60 minutes after the task), and the data obtained before and after the task on each muscle were compared. Comparisons of the normalized data were also performed between the two muscles at each time point. As a result, a significant increase in muscle hardness was seen at 1 minute after the task in the biceps brachii muscle (p=0.0093). In contrast, the masseter muscle showed a tendency to lower hardness, with the lowest point of hardness occurring at 10 minutes after the task (p = 0.0160). Between the two muscles, there was a difference in the normalized data immediately after the task, and at 1, 5, and 10 minutes after the task (0.01 or = 2 h during the previous day. Among the children < or = 10 years 28.7% had a television in their bedroom, among children > 10 years this was 45.7%. Age (> 10 years), ethnicity (notably Surinam origin) and having a television in the bedroom was related to spending more time watching television. Having parents with a high socioeconomic status (SES) was associated with less television viewing than having parents with a lower SES. Children who had not eaten fresh fruit or who had visited a snackbar the previous day had been watching television for > or = 2 h more often than children who had eaten fresh fruit (p < 0.001) or who had not visited a snackbar (p < 0.05). CONCLUSION: Children spent a lot of time watching television. In view of the relation between television viewing and overweight this is an alarming development. Possibilities for the prevention of overweight by reducing television viewing must be investigated. PMID- 15532332 TI - [Turning the head, an unusual mechanism to compensate for diplopia caused by abduction restriction of one eye]. AB - A 59-year-old-man visited the neurological outpatient clinic because of a leftward rotation of his head for the last 8 months. This head deviation turned out to represent a compensatory mechanism to alleviate diplopia that resulted from an abduction restriction of his left eye. By turning his head into the direction of the weak left lateral rectus muscle, the patient could fixate both eyes on target and maintain binocular vision. PMID- 15532333 TI - [Cutaneous myiasis caused by a double infestation with larvae of Dermatobia hominis and Cochliomyia hominivorax]. AB - In a 51-year-old man who had visited Surinam, cutaneous myiasis was diagnosed, caused by simultaneous infestation with the larvae of two different species of flies: Dermatobia hominis and Cochliomyia hominivorax. On his right lower arm the man had two solitary, furuncle-like lesions with a central breathing hole. Two days after these holes had been occluded with vaseline, two white larvae of D. hominis emerged. On both ankles the man had large, undermined ulcers containing hundreds of creeping larvae about 2 cm in length with a salmon-like colour: C. hominivorax. The larvae were removed from the ulcers by hand and by rinsing with physiological saline, after which the wounds healed rapidly. Myiasis is seen in the Netherlands mostly in people returning from a holiday in myiasis-endemic areas. PMID- 15532334 TI - [Notifications to the Netherlands Pharmacovigilance Foundation Lareb of the expulsion of a vaginal contraceptive ring (NuvaRing) and of pregnancy during the use of it]. PMID- 15532335 TI - [Acetylsalicylic acid in patients with diabetes for the primary prevention of cardiovascular disease]. PMID- 15532336 TI - How would military hospitals cope with a nuclear, biological, or chemical disaster? AB - In any nuclear, biological, or chemical (NBC) attack, it is clear that the hospitals may be overwhelmed with casualties. In these deliberate accidents, there will be an additional problem of contamination. Military hospitals must be prepared for NBC disasters with a detailed incident plan. Within this plan, decontamination facilities and shower systems are needed primarily to protect the hospital from self-contamination. Physical and collective protection measures of the staff should be taken into account, with evacuation routes under such an attack. Within this conjunction, the required equipment, including protective suits, detection means, drugs, antidotes, and vaccines, should be provided and stored properly. Qualified personnel should be assigned to the NBC first-aid and rescue team organized within the hospital, equipped, and trained according to such a possible task. Medical staff must be aware of the effects of the agents, and must be experienced in decontamination and first-aid to victims exposed to these agents. Therefore, this information must be put into practice by giving attack scenarios and responses to the hospital administration. It can be conclusively pointed out that military hospitals especially should be better prepared than any other civilian health unit to such attacks of weapons of mass destruction. PMID- 15532337 TI - The recommended role of exposure biomarkers for the surveillance of environmental and occupational chemical exposures in military deployments: policy considerations. AB - A lack of individual exposure information limited the evaluation of exposure outcome relationships after the Gulf War. Exposure concerns during Operation Enduring Freedom and Iraqi Freedom deployments have increased interest in individual environmental and occupational chemical exposure assessment. Currently, deployment assessments are conducted using intermittent ambient air monitoring, occasional focused evaluations based on these results, and postdeployment questionnaire documentation of exposure and/or health concerns. Although this strategy is an improvement over previous practice, it has limitations, including a reliance on evidence of an acute problem, to initiate in depth health evaluation. Exposure biomarkers may have the potential to overcome some of the limitations of current environmental and occupational exposure assessment tools. This article examines current exposure assessment methods, reviews emerging technologies, and recommends a phased approach to introducing exposure biomarkers into a comprehensive occupational and environmental health surveillance program. PMID- 15532338 TI - Neurology in the Vietnam War: CPT Carr's patients. AB - CPT Andrew C. Carr was the second neurologist to be assigned to Vietnam during the early years of the war. Soon after his return, he prepared a commentary on the practice of neurology under field conditions, providing a vivid account of the practice of good neurology under bad conditions. PMID- 15532339 TI - Dental satisfaction survey at an expeditionary dental clinic in support of Operations Enduring Freedom and Iraqi Freedom. AB - This study used a survey to gather information about soldiers' perceptions of the quality of dental treatment delivered at a deployed clinic. An ordinal scale of 1 through 5 scored access, dental facility and equipment, pain control, technical skill, and interpersonal skills of the provider. Four additional items identified whether the respondent was new to military or deployment dentistry, was in pain before arriving at the clinic, or had any concerns about being treated at a deployed dental clinic. Percentages for each type of response were recorded. Data were further analyzed by the Mann-Whitney U test to investigate the relationship between respondent variables and how they rated the attributes of quality. Deployed service members reported a high degree of satisfaction with the quality of care they received. Ninety-seven percent of respondents believed that the care they received was as good as the care they would have received at their regular stateside military dental facility. PMID- 15532340 TI - Cardiovascular risks in a military health care beneficiary population with high blood pressure. AB - Prevention strategies for coronary artery disease among hypertensive patients require assessment of other modifiable risk factors in the target population. In this article, we describe the prevalence of other cardiovascular risk factors in military beneficiaries with high blood pressure (HBP). Baseline data from an ongoing randomized clinical trial designed to test effectiveness of a comprehensive HBP intervention are used in the analysis. A total of 147 beneficiaries from a military health system participated in this study. Findings indicate that the rate of HBP control in this sample was suboptimal (32%: blood pressure < 140/90). Other prevalent cardiovascular risk factors include high cholesterol, obesity, and diabetes. Many of these patients (31.3%) are also in the moderate-to-high danger level of developing coronary artery disease in the near future. These findings warrant a regular assessment of cardiovascular risk factors and rigorous behavioral interventions for all beneficiaries of the military health care system. PMID- 15532342 TI - A multivariate analysis of factors associated with differential disease and nonbattle injury and morbidity aboard ships of the U.S. Naval 5th Fleet during peacetime deployment. AB - Disease nonbattle injury (DNBI) surveillance is a critical component of U.S. military force health protection and has been aggressively implemented by the U.S. Central Command. This study presents a multivariate analysis of factors associated with DNBI incidence rates as well as a description of morbidity measures associated with DNBI from U.S. Navy ships deployed to the Middle East from October 2000 through September 2001. Weekly DNBI reports (N = 331) from a total of 44 individual units representing six different classes of U.S. Navy ships were included in the analysis. There were statistically significant differences in summary and categorical DNBI rates associated with ship class, season, and presence of female sailors embarked. The top three DNBI categories associated with the most lost workdays because of sick in quarters and hospitalization were other medical/surgical (36%), infectious gastrointestinal (23%), and all types of nonbattle injury combined (17%). PMID- 15532341 TI - The causes of false-positive medial branch (facet joint) blocks in soldiers and retirees. AB - OBJECTIVE: The purpose of this study was to determine the factors associated with false-positive medial branch blocks (MBB), the nerve blocks used to diagnose facet arthropathy, in soldiers and retirees. METHODS: The study subjects were 78 patients with chronic low back pain who underwent diagnostic MBB to determine whether or not the facet joints were pain generators. Radiofrequency denervation of these nerves was performed in all patients with positive responses. Patients who failed to obtain pain relief after the blocks (negative blocks) and those who obtained temporary pain relief after MBB but failed radiofrequency denervation (false-positive blocks) then proceeded to undergo discography. Based on patients' responses to diagnostic blocks, discography results, the presence of radicular pain, and previous back surgery, the data were analyzed to determine whether any of these variables correlated with false-positive MBB. RESULTS: The presence of discogenic or radicular pain was not associated with a higher false-positive response rate to MBB. Conversely, the absence of discogenic pain was associated with a higher percentage of false-positive blocks. There was also a trend for patients with previous back surgery to have a higher false-positive rate than those who had not undergone previous surgery. CONCLUSIONS: Although a high incidence of epidural and foraminal spread occurs during the performance of MBB, this is unlikely to be a significant cause of false-positive blocks. PMID- 15532343 TI - National examination of compliance predictors and the immunization status of children: precursor to a developmental model for health systems. AB - OBJECTIVES: The gap between children's immunization requirements and actual compliance is staggering. Using a national survey instrument, the purpose of this study was to analyze the relationship between select predictors and immunization status in children between 19 months and 6 years of age. METHODS: From the 1998 Medical Expenditure Panel Survey, 1575 children were included in a multivariate logistical model. Outcomes were measured as compliance with approved vaccination regimens. RESULTS: Only 67% of children between 19 months and 6 years old were compliant with all recommended vaccines. However, nonminority children with health insurance from families with income above the federal poverty line or living outside a metropolitan statistical area were more likely to be vaccinated. Children of older or educated parents likewise tended to have complete immunization. CONCLUSIONS: Analyzing childhood immunization via parental characteristics identifies prevalent predictors of compliance. Outcomes suggest potential strategies for health systems to focus on children's health issues- specifically immunizations. PMID- 15532344 TI - Prospective validation of the Ottawa Ankle Rules in a deployed military population. AB - This study prospectively validated the Ottawa Ankle Rules (OAR) in patients presenting with acute ankle trauma to a deployed military clinic at Prince Sultan Air Base in Saudi Arabia. The treating physician determined whether the patient met OAR criteria. The decision to obtain radiographs was left to the discretion of the physician. All radiographs were read by a radiologist blinded to the study. Patients who were not evaluated by radiographs received follow-up in the orthopedic clinic or by telephone. Forty-five consecutive patients were enrolled over a 3-month period. Twenty-nine (64%) patients met the OAR criteria, 32 (71%) received radiographs, and 5 (11%) fractures were identified. All fractures were predicted by the OAR. The sensitivity of the OAR was 1.0 and specificity was 0.40. Negative predictive value was 1.0, positive predictive value was 0.17, likelihood ratio positive value was 1.67, and likelihood ratio negative value was 0.0. The OAR correctly predicted all ankle fractures in the military population studied. PMID- 15532345 TI - Orthopedic injuries during Operation Enduring Freedom. AB - Orthopedic injuries comprise a majority of combat injuries seen in recent U.S. military conflicts. Interventions in the forward deployed area have played an important role in improving mortality rates of soldiers as well as outcome at a medical center level. A retrospective review was conducted on orthopedic injuries from Operation Enduring Freedom evaluated at Walter Reed Army Medical Center (WRAMC). Patients were grouped into one of five injury categories (open fracture, amputation, arterial injuries, neurological injuries, and soft tissue injury) with evacuation time (days from time of injury to arrival at WRAMC) and procedures performed before arrival at WRAMC evaluated. The average evacuation time for all orthopedic casualties was 7.9 days. There was an average of 2.6 procedures performed per patient before arrival at WRAMC. There was no difference in evacuation time among the injury groups. Those with only soft tissue injuries underwent fewer procedures than the other injury groups; however, there was no difference among the injury groups in terms of procedures performed. The number of procedures performed did not affect the evacuation time. Fifty-six percent of casualties required operative intervention after arrival at WRAMC. With the unavoidable evacuation time that all casualties must endure regardless of severity of the injury, early operative intervention in forward deployed medical assets, such as the forward surgical team and combat support hospital, remains a necessity for rehabilitative and reconstructive efforts of the soldiers at the medical center level. PMID- 15532346 TI - Lessons learned: Operation Anaconda. AB - With the recent long deployments of our military, a review of lessons learned becomes a vital part of mission success and helps to shape our forces for future conflicts. This article examines some medical issues that were experienced by the Raider Rakkasans of the 101st Airborne (Air Assault) Division during Operation Anaconda in Afghanistan. Topics include nutrition, altitude sickness, medical evacuation requests, and the evacuation of wounded on resupply platforms (back hauling). When nutritional aspects of war fighting were discussed, an emphasis was placed on making a greater variety of field rations accessible to the force with the addition of a multivitamin to each ration. Also, a meal high in caloric content before infiltration and an education/inspection program to ensure the maximum benefit of each ration was reviewed. The use of acetazolamide to combat altitude sickness was also discussed. Dosing of 250 mg BID 24 hours before a rapid assent, then 125 mg BID for 4 days was suggested. Line three of the medical evacuation request was also reviewed for reporting inaccuracies. Complete physical examinations and direct dialogue between medical personnel and commanders was stressed. Last, a standard medical package to accompany any resupply during hostilities was advised, instead of releasing ground medical personnel to attend the wounded when using this for an evacuation platform. PMID- 15532347 TI - Psychosocial effects of the BOOT STRAP intervention in Navy recruits. AB - The purpose of this prospective study was to investigate the effects of the Boot Camp Survival Training for Navy Re cruits--A Prescription (BOOT STRAP) intervention on stress depression, situational events, interpersonal factors, and recruit training performance. Divisions of Navy recruits were randomly selected and 801 recruits participated for the 9 weeks of their training. Recruits "at risk" for depression were randomly assigned to the intervention or nonintervention groups, and the remaining recruits served as the comparison group. The at-risk recruits who received the BOOT STRAP intervention significantly increased their sense of belonging, experienced less loneliness, used more problem-solving coping skills, and decreased insecure attachment by the end of recruit training. Percentages of recruits in the study success fully completing basic training were 84% of the comparison group, 86% of the intervention group, and only 74% of the nonintervention group. Results suggest that the BOOT STRAP intervention improves recruit functioning, strengthens train ing performance, helps reduce attrition, and may have impor tant implications for stress and depression interventions. PMID- 15532348 TI - Unmasking of carnitine palmitoyltransferase deficiency during an acute exacerbation of asthma complicated by rhabdomyolysis in a soldier. AB - Rhabdomyolysis is a life-threatening condition that may result from various etiologies. We report a rare case of severe rhabdomyosis in a soldier after a mild acute asthma exacerbation. Further work-up revealed an underlying deficiency of type II carnitine palmitoyltransferase. The case is discussed along with a review of the literature. It is concluded that acute asthma exacerbations may be a unique precipitating factor of rhabdomyolysis and may therefore unmask underlying metabolic myopathies. Asthma may cause rhabdomyolysis through several different mechanisms, and thus the occurrence of rhabdomyolysis in the context of asthma exacerbations should warrant a work-up for metabolic diseases, especially in the presence of high creatine kinase levels. Given the high incidence of asthma, especially among young adults, a high index of suspicion is needed in order that rhabdomyolysis be promptly diagnosed and treated. PMID- 15532349 TI - Management of lower back pain in young Turkish recruits. AB - The aims of this study were to determine the appropriate management strategy for lower back pain in the young male population of the Turkish Army and to analyze the results of surgical treatment of such a common health problem versus medical treatment. Participants in the study included 865 recruits who underwent basic military training between June 2001 and June 2002. All were admitted to our department with the complaint of lower back pain and were evaluated by neurological and radiological examinations. We operated on 23 of them and followed-up by 3 months. We compared the data of surgically and nonsurgically treated patients. In our opinion, the indications for surgery for lower back pain must be strict, and careful selection of patients is needed for a satisfactory outcome. PMID- 15532350 TI - Did substance use change after September llth? An analysis of a military cohort. AB - The purpose of this study was to assess changes in alcohol, tobacco, and other drug use after the terrorist attacks of September 11 among a cohort of young military personnel consisting of 661 active duty Navy personnel, including 164 women. Paired sample t tests were computed to assess mean differences in substance use in the 30 days before and after September 11. Average daily alcohol consumption significantly declined in the 30 days after September 11 among enlisted personnel (n = 315) and officers (n = 360). Significant increases in the usual number of cigarettes smoked and in the number of days using prescription drugs after September 11 were observed among enlisted personnel, but not among officers. Increased use of cigarettes and prescription drugs among enlisted personnel after September 11 suggests that some sectors of the military may turn to tobacco and other substances to cope with traumatic circumstances such as the events of September 11. PMID- 15532352 TI - Stress fracture of the ulna occurring in military recruits after rifle drill training. AB - Stress fractures are a common injury in military recruits in the lower extremities, but are rare in the upper limbs. The locations of reported stress fractures occurring in the upper extremities are mainly in the ulna and olecranon. We report on two cases of mid-ulnar stress fractures in a team of honor guards following rifle drill training from excessive weight lifting, repetitive pronation of the forearm, and a sudden increase in the training load. According to the report, some readjustment must be made in the training protocol for military recruits. Stress fractures are also being recognized as a prevalent problem in the upper body. PMID- 15532351 TI - Health effects of anthrax vaccination in the Canadian forces. AB - OBJECTIVE: The objective of this study was to determine whether anthrax vaccine resulted in adverse health effects in Canadian Forces members 8 months after vaccination. METHODS: A quasi-experimental, retrospective chart review was undertaken for two groups within the Canadian Forces, one group that received anthrax vaccination and another that did not. Information on symptoms, diagnoses, and injuries for 848 persons for which there were approximately 35,000 chart entries was abstracted from charts over a 4.5-year period and was coded using the International Statistical Classification of Diseases and Related Health Problems, 10th edition. RESULTS: The chart retrieval rate was 84%. The mean number of chart entries per person was higher in the comparison group (43.4) than in the vaccine group (38.2). No statistically significant differences were seen in the percent change before and after vaccination in the number of chart entries for specific diagnoses and symptoms for the vaccine group compared with the comparison group. Visual inspection of the time trend in rates showed no unexplained increases in the rate of diagnosis and symptoms in the vaccine group after vaccination. CONCLUSION: This study found no evidence that the anthrax vaccination resulted in an increase in adverse health effects in the 8-month period after vaccination. PMID- 15532353 TI - Cerebral oximetry (INVOS technology). PMID- 15532354 TI - Neuromotor issues in human performance: introduction. AB - Two current approaches frequently implemented to understand mechanisms involved in motor control are presented in a series of papers. One approach aims at understanding what specific neural mechanisms are responsible for motor behavior Such neural mechanisms might be operative at any of several sites in the nervous system, including spinal sites capable of complex reflex organization, as well as supraspinal sites involved in integrating multiple sensory and motor processes. Alternative approaches involve the assessment of kinematic and other variables used to describe the behavior using dynamical systems theory. Such an approach allows the researcher to explore relationships among groups of variables in, for example, multijoint control. In the series of papers that follow, the value of both of these approaches is discussed using examples from tasks that involve both rhythmic activities (running and cycling) and discrete movement (weight lifting and golf). PMID- 15532355 TI - Neural issues in the control of muscular strength. AB - During the earliest stages of resistance exercise training, initial muscular strength gains occur too rapidly to be explained solely by muscle-based mechanisms. However; increases in surface-based EMG amplitude as well as motor unit discharge rate provide some insight to the existence of neural mechanisms in the earliest phases of resistance exercise training. Moreover, other phenomena such as the cross transfer of strength following unilateral exercise and neural changes following limb immobilization also support a role for neural control in muscular strength. While studies to date have focused on the phenomenon of neural change, future studies will need to focus on the many motoneuron influences as well as intrinsic motoneuronal properties that may be responsible for these neural adaptations. PMID- 15532356 TI - Neuromotor issues in the learning and control of golf skill. AB - Theoretical and practical issues related to the neuromotor control of a golf swing are presented in this paper. The typical strategy for golf training consists of high volume repetition with an emphasis on a large variety of isolated swing characteristics. The student is frequently instructed to maintain consistent performance in each swing with absolute invariance. Based on dynamical systems and motor control schema perspectives, it is argued that golfers can learn a more reliable swing by exploring swing parameters and focusing on higher order control principles that reduce the vast number of degrees of freedom. Some candidate training practices are proposed for applying these theoretical issues into practice. PMID- 15532357 TI - Neuromuscular control and coordination during cycling. AB - The neuromuscular control aspect of cycling has been investigated through the effects of modifying posture and cadence. These studies show that changing posture has a more profound influence on neuromuscular coordination than does changing slope. Most of the changes with standing posture occur late in the downstroke: increased ankle and knee joint moment, reduced hip joint moment and greater activity in specific muscles. Due to the influence of lower extremity inertial properties, higher pedaling frequency induces more neuromuscular changes at the hip than at the knee or ankle joints. These neuromuscular adaptations to environmental and task constraints are discussed with regard to the contributions of the central nervous system and the solution provided by peripheral anatomical structure--mono- and biarticular muscles. The results indicate that training and related movement analysis should be specific to the motion, supporting the notion of task-specific training. PMID- 15532358 TI - Neuromuscular characteristics of endurance- and power-trained athletes. AB - In response to chronic physical training, the human neuromuscular system undergoes significant and specific adaptations. More importantly, these influences are the result of the type and quantity of physical activity. One of the simplest neuromuscular mechanisms is the spinal stretch reflex. The reflex system was previously viewed as inflexible, with a relatively fixed response that could vary only slightly. However, more recent data have identified an adaptive plasticity in the reflex system. In this respect, the reflex system can be used to assess training and aging adaptations of the human neuromuscular system. Due to their methodological simplicity, both the tendon-tap reflex and the electrically evoked Hoffmann reflex (H-reflex) can be used to assess training adaptations of the human neuromuscular system. The purpose of this paper is to review briefly the tendon-tap and H-reflex paradigms and delineate the research findings pertaining to changes in the reflex system with physical training. For purposes of clarity, this discussion will be divided into the following: (a) differences observed in the tendon-tap reflex, (b) differences observed in the H reflex, and (c) role of interneurons in mediating these changes. PMID- 15532359 TI - Gender differences among sagittal plane knee kinematic and ground reaction force characteristics during a rapid sprint and cut maneuver. AB - Women are more prone to anterior cruciate ligament (ACL) injury during cutting sports than men. The purpose of this study was to examine knee kinematic and ground reaction forces (GRF) differences between genders during cutting. Male and female athletes performed cutting trials while force platform and video data were recorded (180 Hz). Differences (p < . 05) were observed between groups for knee flexion at contact and GRF at maximum knee flexion. Women averaged 5.8 degrees less flexion at contact and 1.0 N. (kg x m x s(-1))(-1) greater GRF at maximum flexion. Knee range of motion and peak GRF variables were not significantly different, but women had greater values. Women exhibited technique characteristics believed to increase ACL injury risk, but men exhibiting similar characteristics were also observed and could also be at risk. PMID- 15532360 TI - Effects of repeated retention tests on learning a single timing task. AB - The purpose of the present study was to investigate the possibility that administering an initial retention test would influence any subsequent retention tests administered to the same participants. Participants performed 40 practice trials of a four-segment key-pressing task with a movement time goal of 925 ms. Participants were then administered either two retention tests (Day 1-Day 1 group, 10 min and 20 min after practice; Day 1-Day 2 group, 10 min and 24 hr after practice; Day 2-Day 2 group, 23 hr 50 min and 24 hr after practice) or one retention test (Day 1-Control group, 20 min after practice; Day 2-Control group, 24 hr after practice). The first test had a deleterious effect on performing the second test for the Day 2-Day 2 group, which would be expected if the task representation was updated and re-encoded at the time of recall using diminished information. This finding suggests that researchers should be cautious about using repeated retention tests that are administered 1 day after practice. In addition, teachers and coaches should consider administering a test under gamelike conditions following practice of a new technique. Otherwise, performance of the new technique may be degraded or perturbed at the time of later performances. PMID- 15532361 TI - What determines limb selection for reaching? AB - While motor dominance appears to drive limb selection for reaching movements at the midline and ipsilateral (dominant) side, this study examined the possible determinants associated with what drives the programming of movements in response to stimuli presented in contralateral space. Experiment 1 distinguished between object proximity and a hemispheric bias for using the hand on the same side as the stimulus by comparing imagined and actual reaching with arms uncrossed and crossed. Experiment 2 examined the role of comfort in limb selection via participants reaching for a cube under two conditions: 1- and multiple-df movements. The first experiment demonstrated a proximity effect; participants preferred to keep their arms crossed to reach with the hand closest to the stimulus rather than uncrossing their arms to support a bias. Comfort was eliminated when participants continued to prefer the nondominant hand, even though they were able to reach with the dominant hand at all positions using only a 1-df movement. In summary, we offer evidence suggesting that limb selection in response to stimuli presented in contralateral hemispace is determined primarily by information related to the proximity between the stimulus and the hand. PMID- 15532362 TI - Expert golf instructors' student-teacher interaction patterns. AB - The purpose of this study was to identify the dominant instructional interaction patterns of expert golf instructors. Instructors (N = 22) were selected by the Ladies Professional Golf Association (LPGA) Teaching based on the following criteria: (a) 10 or more years of golf teaching experience, (b) LPGA certification, (c) awards received for the quality of their instruction, and (d) peer and student recognition for outstanding teaching. The instructors were videotaped teaching a 60-min lesson to a novice college-age woman with no previous golf experience. The tapes were then analyzed using both the Cheffers Adaptation of Flanders' Interaction Analysis System (CAFIAS) and a qualitative analysis. Based on the findings from descriptive statistics and correlation analyses of the CAFIAS data and qualitative data analysis, several trends were identified. First, the dominant instructional behavior of these teachers was providing information to the students using both explanations and demonstrations. Second, the prevailing instructional interaction pattern of the expert teachers included extensive explanations and demonstrations followed by directions. The student followed the directions by practicing skills and received praise for their achievements. Third, high rates of directions and praise from teachers prompted student practice. Fourth, engaging students in subject-related discussion was positively correlated with teachers' questions but negatively correlated with teachers' criticisms. Finally, teacher acceptance was positively correlated with student analytic behavior, while teachers' talk negatively correlated with students initiating discussions. PMID- 15532363 TI - Children's motivation in elementary physical education: a longitudinal study. AB - The present study examined relationships among variables drawn from achievement goal theory and the expectancy-value model of achievement choice as well as mean level changes of these variables over time in elementary physical education. Participants (N = 207) completed questionnaires over a 2-year period: once while in the second and fourth grades and again when they were in the third and fifth grades. Results indicated that achievement goals, expectancy-related beliefs, and subjective task values were related to one another and were predictive of children's intention for future participation in physical education. Children's subjective task values of physical education decreased over time. Children in Cohort 1 (across second to third grades) generally had stronger motivation for learning in physical education than children in Cohort 2 (across fourth to fifth grades). Findings suggest the importance of integrating achievement goal theory and the expectancy-value model of achievement choice in understanding student motivation. PMID- 15532364 TI - Relationships between exercise regulations and motivational consequences in university students. AB - The aim of this study was to examine propositions stemming from Self Determination Theory (SDT) that contend motivational consequences vary as a function of different regulations in the exercise domain. Participants (N = 276; n = 98 men; n = 178 women) completed inventories assessing exercise regulations, current exercise behavior, and behavioral intentions to continue exercising for the next 4 months and the effort and importance associated with exercise participation. Bivariate analyses indicated autonomous exercise regulations (identified and intrinsic) were the strongest correlates of each motivational consequence across both sexes, and introjected regulation was positively associated with exercise consequences in women. Simultaneous multiple regression analyses indicated that exercise regulations accounted for a sizeable portion of the variance across each motivational consequence in both sexes (R2adj values ranged from .20 to .53). Both regression and structure coefficients revealed that introjected regulation was a stronger motivational force in women than men, and identified regulation was the most important predictor of all three motivational consequences in both sexes. Collectively, these findings suggest that exercise regulations differentially predict motivational consequences across sexes, and future research using this theoretical framework for examining motivational issues pertinent to the exercise domain appears warranted. PMID- 15532365 TI - Cross-cultural comparison of American and Finnish college students' exercise behavior using transtheoretical model constructs. AB - Although the benefits of exercise are well documented, an international problem of physical inactivity exists. More research, especially theory based, has been recommended. One promising approach for studying exercise behavior is that proposed in the Transtheoretical Model (TTM) of behavior change. This model, however, has received minimal cross-cultural attention and, relative to the current study, measurement instruments have only recently been translated into the Finnish language. The purpose of this study was to assess American and Finnish college students' exercise behaviors on the basis of TTM. Participants were American (n = 169) and Finnish (n = 168) college students who completed language-specific measures of exercise behavior, stage of change, processes of change, decisional balance, self-efficacy, and temptation. The only cultural difference observed was that the American participants rated themselves higher on barrier self-efficacy relative to the Finnish participants. Regardless of nationality orgender, participants classified by their stage of change differed on all the core constructs assessed. These results generally support the utility of TTM for understanding American and Finnish college students' exercise behavior. PMID- 15532366 TI - Time course of attention and decision making during a volleyball set. PMID- 15532367 TI - Attraction to physical activity mediates the relationship between perceived competence and physical activity in youth. PMID- 15532368 TI - [Chronic progressive nephropathies and modern lifestyle]. PMID- 15532369 TI - [Arterial hypertension in chronic glomerulonephritis: detectability and treatment efficacy]. AB - AIM: To study prevalence of arterial hypertension (AH) in patients with chronic glomerulonephritis (CGN), its relationship with activity of the renal process, renal function; to analyse policy and efficacy of antihypertensive therapy. MATERIAL AND METHODS A total of 250 CGN patients treated in 1993-2001 participated in the trial. They had different morphological variants of CGN. AH was diagnosed in 193 patients. In the course of the trial changes in antihypertensive treatment policy were observed. RESULTS: AH was most prevalent in mesangiocapillary (96.6%) and diffuse fibroplastic nephritis (83.9%). In functional insufficiency of the kidneys AH occurred in 90.1%. AH was associated with clinical and morphological signs of nephritis activity, severity of tubulointerstitial alterations, purin and lipid metabolism. Uric acid level and age were independent prognostic factors of AH development. AH correction was achieved in the initial and subsequent periods in 51.7 and 58.7% cases. Later, ACE inhibitors were prescribed more often, both in monotherapy and in combination with other drugs; calcium antagonists were taken less frequently. CONCLUSION: AH in CGN patients is a frequent finding and depends on a morphological nephritis variant, activity of the renal process and degree of renal failure. Age, gender and metabolic disorders are also involved in AH development in CGN patients. Recently, there is a trend to more frequent prescription of combined treatment. Drugs of choice in the treatment of renal AH are ACE inhibitors. PMID- 15532370 TI - [Association of the complex of polymorphic markers of ACE genes, aldosteron synthetase and endothelial synthetase of nitric oxide with progression of chronic glomerulonephritis]. AB - AIM: To study association of the complex of polymorphic markers of ACE genes (ACE complex), aldosteron synthetase gene (CYP11B2) and endothelial synthetase of nitric oxide (NOS3) with onset, course and progression of chronic glomerulonephritis (CGN). MATERIAL AND METHODS: 117 CGN patients were examined. Genetic predisposition to CGN development was studied by comparison of distributions of alleles and genotypes of polymorphic markers of genes ACE, CYP11B2 and NOS3 in CGN patients and controls (n = 80) free of renal diseases and arterial hypertension (AH). The course of CGN was analysed with consideration of the following factors: AH severity, proteinuria persistence, nephritic level for 6 months and longer, immunosuppressive therapy and response to it, therapy with ACE inhibitors and/or blockers of antiotensin II receptors (ARB). CGN progression rate end point was doubling of initial blood creatinine level. RESULTS: Significant differences in the incidence of the above alleles and genotypes in the patients and controls were not found. The patients were divided into two groups: group 1 consisted of 25 patients carrying the combination of alleles D+C+4a, group 2 consisted of the rest 92 patients. The groups did not differ by CGN course parameters, but renal survival was significantly lower in carriers of the allele combination D+C+4a. Cox's mono- and multifactorial regression analysis has shown that carriage of the allele combination D+C+4a is an independent riskfactor of renal survival deterioration. CONCLUSION: No association was detected between polymorphic markers of genes ACE, CYP11B2 and NOS3 and onset of CGN. Carriage of D+C+4a allele combination is an independent factor of risk for fast progression of chronic renal failure. PMID- 15532371 TI - [Chronic glomerulonephritis and pregnancy]. AB - AIM: To clarify factors of risk for unfavourable variants of gestational chronic glomerulonephritis (CGN) and poor pregnancy outcomes in CGN; to determine prognostic implications of changes in some renal and uteroplacental indices. MATERIAL AND METHODS: Variants of CGN gestational course and pregnancy outcomes have been analysed for 156 CGN patients. The women were examined before pregnancy, in the course of pregnancy and 3-24 months after the delivery. Measurements were made of 24 h proteinuria, glomerular filtration rate, blood transaminases activity, functional renal reserve (FRR), uricemia, blood level of alpha-phetoprotein. Placentas were studied morphologically, uterine and umbilical artery circulation was assessed by dopplerometry. RESULTS: The following abnormalities were registered: high proteinuria (34.6%), progression of hypertension (29.5%), renal function deterioration (15.4%), fetal and neonatal losses (15.4%), fetal underdevelopment (25%), preterm delivery (17.3%), preeclampsia (7.7%), preterm placental detachment (1.9%). There is morphological, dopplerometric and biochemical evidence for placental insufficiency in CGN pregnant women. CONCLUSION: Activity of CGN (nephritic or acute nephritic syndromes), hypertension, renal failure, disorders of renal hemodynamics are factors of risk for unfavourable gestational course of CGN and pregnancy complications. Placental insufficiency deteriorates pregnancy outcomes in CGN, but changes in uterine and umbilical circulation as well as blood levels of alpha fetoprotein are not prognostically significant. PMID- 15532372 TI - [Cyclophosphamide efficacy in maintenance therapy of nephrotic syndrome in patients with chronic glomerulonephritis]. AB - AIM: To study effects of cyclophosphamide (CP) on duration of remission in chronic glomerulonephritis (CGN) with nephrotic syndrome (NS). MATERIAL AND METHODS: Achievement of complete remission of NS was followed by intravenous treatment with CP in 25 of 28 patients. 20 patients were in remission for 7-90 months (mean 38.9 +/- 5.6 months). NS recurred in 8 patients after remission duration from 3 to 48 months (mean 25.4 +/- 6.1 months). RESULTS: Patients in remission and NS recurrence were comparable by morphology of CGN, gender, CGN history, start of CP treatment, accumulated CP dose at remission induction, 24 h protein urine loss, levels of immunoglobulins, C-reactive protein, fibrinogen, seromucoid, complement titer, circulating immune complexes. Significant differences were observed in duration of CP maintenance, accumulated dose within the first three months and maintenance therapy. CONCLUSION: CP maintenance in CGN with NS should be conducted as an intravenous pulse-therapy once in 3 weeks for 3 months than once in 1-3 months for 1.5-2 years. PMID- 15532373 TI - [Early diagnosis of renal damage in hypertensive patients]. AB - AIM: To determine early criteria of renal affection in hypertensive patients for improvement of diagnosis and treatment of hypertensive nephropathy. MATERIAL AND METHODS: 148 patients with essential hypertension (EH) were examined (82 males, 66 females, mean age 43.64 +/- 11.35 years). Of them, 46 patients (24 young males among them) had EH of new onset (NOH) and 102 patients had EH of long duration (LDEH). The examination included investigation of lipid, carbohydrate, uric acid metabolism, renal function, microalbuminuria, intrarenal hemodynamics (color dopplerography on ALOKA SSD-2000 MultiView. RESULTS: 50% of NOH and 90% of LDEH patients had metabolic disorders: excessive body weight, hypercholesterolemia, hypertriglyceridemia, uric acid disbolism, impaired glucose tolerance. These disorders, except uric acid metabolism, correlated in severity with duration and severity of EH. Intensity of microalbuminuria depends on EH duration, severity of concomitant metabolic defects. Markers of early renal damage in EH patients are increased intrarenal peripheral vascular resistance and microalbuminuria. These alterations are detectable in 30% NOH patients (at the disease onset). Treatment with ACE inhibitors eliminates microalbuminuria, diminished hyperfiltration and improved intrarenal circulation. This justifies use of ACE inhibitors for nephroprotection in EH patients. CONCLUSION: Monitoring of the above early markers of renal damage and metabolic shifts is essential for assessing progression of hypertensive nephropathy and control over efficacy of antihypertensive treatment including ACE inhibitors. PMID- 15532374 TI - [Low-molecular heparins in the treatment of APS-nephropathy in primary and secondary antiphospholipid syndrome]. AB - AIM: To evaluate efficacy of low-molecular heparin enoxaparin in the treatment of nephropathy in patients with primary and secondary antiphospholipid syndrome (APS). MATERIAL AND METHODS: A total of 11 APS-nephropathy patients participated in the study. Of them 8 patients had primary APS and 3 patients had secondary APS. Biopsy of the kidney was made in 5 patients, ultrasonic dopplerography of renal vessels was performed in all 11 patients. Enoxaparin was given as monotherapy to primary APS and combined treatment to secondary APS patients. RESULTS: Enoxaparin decreased proteinuria, arterial pressure, glomerular filtration rate in all the patients. It also promoted normalization of creatinin. Before the anticoagulant treatment a reduction of maximal systolic and diastolic velocities of blood flow have been registered in segmental and interlobar arteries in all APS nephropathy patients compared to healthy subjects. The depletion of renal cortex blood flow beginning with the level of arcuate arteries was also observed in all the patients. Enoxaparin enhanced blood flow in segmental and interlobar arteries. The recovery of blood flow in the arcuate arteries was registered in 6 of 9 patients. CONCLUSION: Low molecular heparin may be used in the treatment of nephropathy in patients with primary and secondary APS. PMID- 15532375 TI - [Cardioprotective effect of erythropoietin preparations in patients with chronic renal failure]. AB - AIM: To investigate effects of early correction of anemia on the rate of cardiovascular complications and survival on regular hemodialysis (RHD). MATERIAL AND METHODS: Eighty patients with chronic renal failure (CRF) on regular hemodialysis entered two groups: group 1 with hemoglobin (Hb) < 80 g/l (n = 36) and group 2 with Hb > 100 g/l (n = 44). 90% patients of group 2 were treated for renal anemia for 6-8 months of predialysis CRF. When placed on RHD, group 1 started therapy with epoetin, 39 patients of group 2 continued epoetin treatment. RESULTS: Patients of group 2 had a higher rate of eccentric left ventricular hypertrophy (LVH) with reduced ejection fraction and development of congestive cardiac failure and coronary heart disease. Eccentric LVH in group 1 patients regressed only in 80% when the patients were on hemodialysis and received epoetin for correction of anemia. Overall cardiac death in group 1 was twice that of group 2 patients. CONCLUSION: Early correction of anemia led to a 50% increase in 5-year survival. This fact can be explained with inhibited progression of eccentric LVH. PMID- 15532376 TI - [Impact of a hemodialysis session on cardiac function in patients with chronic renal failure]. AB - AIM: To determine the impact of hemodialysis (HD) session on cardiac function in patients with chronic renal failure. MATERIAL AND METHODS: Thirty patients (17 male, 13 female, mean age 49 +/- 11 years) on bicarbonate HD were studied. M-mode echocardiography was performed and ejection fraction (EF) was estimated. Transmitral flow was assessed by Doppler echocardiography. Peak velocity of early (E) and late (A) filling, E/A ratio, isovolumic relaxation time (IVRT) and early deceleration time (DT) were estimated. All the estimations were made one hour before and immediately after HD by one investigator. Flow propagation velocity of early diastolic flow was assessed by color M-mode Doppler echocardiography. RESULTS: A significant decrease of the ejection fraction (delta EF) was observed only in patients with intradialytic hypotension. Hemodialysis resulted in a decrease of early flow velocity from 99.2 +/- 23.8 to 80.6 + 26.0 cm/s (p = 0.0000) and E/A ratio from 1.23 +/- 0.57 to 0.98 +/- 0.43 (p = 0.006). IVRT and DT showed no significant difference. There was a significant positive correlation between the amount of ultrafiltration and deltaE (r = 0.46; p = 0.01), there was no correlation between the amount of ultrafiltration and delta Vp (r = -0.01; p = 0.9). CONCLUSION: The results show that a hemodialysis session influences cardiac function in patients with chronic renal failure. Early diastolic filling considerably decreased in correlation with ultrafiltration. A significant decrease in an ejection fraction was detected only in patients with intradialytic hypotension. Ultrafiltration had no impact on flow propagation velocity of early diastolic flow of the left ventricle assessed by color M-mode Doppler echocardiography. PMID- 15532377 TI - [A renoprotective effect of enalapril in chronic transplantation nephropathy]. AB - AIM: To study safety and efficacy of ACE inhibitor enalapril in chronic transplantation nephropathy (CTN) as well as nephroprotective efficacy of this drug in various clinical variants of CTN. MATERIAL AND METHODS: A retrospective study covered 220 recipients with CRF. The patients were divided into the study group (n = 103) and the control group (n = 117). The study group was given ACE inhibitor enalapril the efficacy of which was assessed by arterial pressure (systolic, diastolic, mean) dynamics, 24 h proteinuria and the rate of CTN progression. This rate was suggested by probability of plasm creatinin doubling (Kaplan-Meier technique). RESULTS: Enalapril significantly inhibited CTN progression running with minimal or marked proteinuria, had a pronounced hypotensive effect, promoted stabilization of minimal proteinuria (in CTN with minimal proteinuria) or reduction of protein excretion (in a proteinuric variant of CTN). CONCLUSION: Use of enalapril in CTN in a daily dose 10 mg maximum is safe and can be used for inhibition of CTN progression. PMID- 15532379 TI - [Epidemiology of chronic renal disease in the North-West of Russia: setting-up the register]. AB - AIM: To study epoidemiology of chronic renal disease (CRD) in the North-West region of the Russian Federation. MATERIAL AND METHODS: A population retrospective trial was performed from 01.07.98 to 30.06.99 in the Vologda region, towns Veliky Novgorod and Syktyvkar (total population 1840000). 490 cases of chronic renal failure (CRF) with creatinin levels > 300 mcmol/l corresponding to CRD stage IV and V by glomerular filtration rate were detected. CRD stage IV-V prevalence, morbidity including sex- and age-specific parameters, cumulative survival in different etiology of CRD regarding replacement therapy (RT) and nephrologist supervision were estimated. RESULTS: Mean prevalence and morbidity were 266 and 108 cases per million. Number of affected males and females was the same. CRD was provoked by chronic glomerulonephritis (CGN)--41%, chronic interstitional nephritis--16%, diabetes mellitus--14%, renal polycystic disease- 8%, amyloidosis--6%, hypertension nephrosclerosis--2%, other causes--3%, cases of unclear etiology--10%. Among patients with primary renal pathology on RT, number of CGN patients was higher--61% while diabetic and amyloidosis shares were much lower 1.6 and 0.8%. 31.1% patients given no RT had Cr > 500 mcmol/l and glomerular filtration rate < 10 ml/min. Incidence of CRD stage IV-V in RT patients was 80 per million, 8 patients per million for 1 year started RT for the first time. CRF mortality was 85 patients per million a year. 88.4% of deaths in patients given no RT was due to uremia. Cumulative survival for RT patients and RT untreated was 90 and 50% a year, respectively. Survival of the RT untreated observed by the nephrologist was significantly higher (p = 0.003). CONCLUSION: New cases of CRD stage IV-V were comparable in number to European incidence rate while prevalence was much less because of lower number of old patients and RT treated. The problem of CRD treatment is urgent for RF and requires updating nephrological service and development of RT programs. PMID- 15532378 TI - [Clinical and biochemical manifestations of chronic pyelonephritis and their correction in population of the northen areas of the Tyumen Territory]. AB - AIM: To study clinicobiochemical features of chronic pyelonephritis (CPN) in the population of northen areas of the Tyumen Territory; to determine emoxipine effects on CPN treatment efficacy. MATERIAL AND METHODS: Clinicolaboratory manifestations of CPN were studied in 246 CPN patients (126 residents of the Tyumen South and 120 residents of the Tyumen North, age 18-55, 174 females, 72 males). The control group consisted of 60 healthy subjects of the same gender and age. RESULTS: CPN in patients living in the north was characterized by more severe intoxication, dysuric disorders, piuria, arterial hypertension, more intensive restructuring of cellular membranes. Membranopathological changes depend on the phase of the pathological process and clinical form of the disease. Emoxipin addition to combined therapy of CPN promoted faster correction of basic CPN symptoms and stabilization of the lipid phase of erythrocytic membranes. CONCLUSION: Clinical and biochemical features of CPN in patients living in the Tyumen North represent additional diagnostic criteria of the disease activity. Adjuvant use of the antioxidant drug emoxipin produces a noticeable clinicobiochemical effect. PMID- 15532380 TI - [Effects of carvedilol, atenolol and their combination with fosinopril on cardiac rhythm variability, clinicofunctional status and quality of life in patients with postinfarction left ventricular dysfunction]. AB - AIM: To study effects of carvedilol and atenolol and their combination with fosinopril on heart rhythm variability (HRV, clinicofunctional status and quality of life in postmyocardial infarction patients with moderate chronic cardiac failure (CCF). MATERIAL AND METHODS: An 8-week randomized open study enrolled 50 male patients (mean age 55.7 +/- 1.58 years) with postinfarction CCF. They were divided into two equal groups. Group one received carvedilol (24.4 +/- 2.0 mg/day) followed by addition of fosinopril (2.5-40 mg/day). Group 2 received atenolol (44.1 +/- 5.1 mg/day) followed by fosinopril in the same dose as in group 1. RESULTS: A 4-week therapy with carvedilol and atenolol effectively corrected depression of HRV in both the group. Combined therapy with these beta adrenoblockers and fosinopril improved impaired global left ventricular contractility, exercise tolerance, quality of life, relieved symptoms of CCF. CONCLUSION: Carvedilol, atenolol and their combination with fosinopril in patients with postinfarction CCF improve clinico-hemodynamic and functional status, raise HRV and quality of life. PMID- 15532381 TI - [Progressive azotemia provoked by ACE inhibitor in renal ischemia]. PMID- 15532382 TI - [Management of patients with diabetes mellitus with terminal renal failure on programmed hemodialysis]. PMID- 15532383 TI - [Dyslipidemia and chronic progressive diseases of the kidneys]. PMID- 15532384 TI - [Risk factors: microalbuminuria]. PMID- 15532385 TI - [Role of the study of urinary molecular mediators of immune inflammation and renal fibrosis in chronic glomerulonephritis]. PMID- 15532386 TI - [Variants of renal affection in Wilson-Konovalov disease]. PMID- 15532387 TI - [Renal damage in antiphospholipid syndrome]. PMID- 15532388 TI - [Primary nursing in ambulatory nursing]. PMID- 15532389 TI - [Time management in the nursing profession]. PMID- 15532390 TI - [Competence profiling for ward administration]. PMID- 15532391 TI - [Critical nutritional deficiencies]. PMID- 15532392 TI - [Good food in the hospital--the newest position?]. PMID- 15532393 TI - [Career profile of nurses]. PMID- 15532395 TI - [Education: Bachelor of nursing]. PMID- 15532394 TI - [Wound care in current hard times]. PMID- 15532396 TI - [The connection with patient files]. PMID- 15532397 TI - [Patient discharge]. PMID- 15532398 TI - [Toward cheap jobs in nursing]. PMID- 15532399 TI - [Continuing education through emotional intelligence]. PMID- 15532400 TI - [One glance makes contact]. PMID- 15532401 TI - [The employment situation]. PMID- 15532402 TI - [Career-overlapping work claims]. PMID- 15532403 TI - [Theories and design is not nursing]. PMID- 15532404 TI - [Transplant ok--patient ko?]. PMID- 15532405 TI - [Discontinuing immunosuppression due to annoying hirsutism. Noncompliance is the number 1 etiology of graft rejection]. PMID- 15532406 TI - [Only 10-30% of smokers achieve long-term abstinence. How do you motivate your patients to abstain from nicotine? (interview by Dr. Thomas Meissner)]. PMID- 15532407 TI - [At first only headache and fever, then neurologic symptoms. Suspected CNS infect: immediate admission to the clinic!]. PMID- 15532408 TI - [Adynamic status, arthralgia, amenorrhea: the 3 "A"s of hemochromatosis]. PMID- 15532409 TI - [2 out of 3 cardiac patients are undetected diabetics. Screening with the oral glucose tolerance test!]. PMID- 15532410 TI - [In memory of Walter M. Gallmeier. Only providing standards of excellence]. PMID- 15532411 TI - [Every second person over 65 years of age has a problem with the thyroid]. PMID- 15532412 TI - [Subclinical hypothyroidism--does it have to be treated?]. AB - Every comprehensive laboratory evaluation should include measurement of the TSH level, since even subclinical hypothyroidism may have a negative impact on the organism. In 95% of all persons with a healthy thyroid, the TSH is between 0.4 and 2.5 microU/ml. If the value exceeds the latter figure, autoimmune thyroiditis as the cause of the subclinical hyperthyroidism must be excluded. Consideration must, however, be given to the fact that severe general diseases, as well as various medications, may have an unspecific influence on the TSH value. Subclinical hypothyroidism is associated with changes in the cholesterol level, cardiovascular effects and neuromuscular symptoms. Short-term studies have demonstrated a positive effect on these symptoms for early replacement treatment with L-thyroxine. PMID- 15532413 TI - [Diminished TSH level--an incidental finding]. AB - Too-low a TSH level detected fortuitously during a routine examination should raise the suspicion of subclinical hyperthyroidism. The question as to whether treatment is required largely depends on the age of the patient and on his general state of health. If considered necessary, treatment is oriented to the guidelines for the treatment of hyperthyroidism. PMID- 15532414 TI - [Medical treatment of nodular goiter]. AB - An optimal supply of iodine--150-200 microg/day--has not yet been secured in all regions and in all phases of life. The appreciable diminishment in thyroid gland mass in children and adolescents is, however, indicative of an improvement in this situation. In elderly persons, however, prophylactic measures do not result in struma or nodule regression. Nodules are found in some 40% of women and 28% of men older than 45. Medical treatment of nodular goiter is aimed at achieving a reduction in size or preservation of the status quo. Depending on the size of the goiter, iodine-deficiency goiter is treated with iodine or a combination of iodine and levothyroxine. In view of the pathogenesis and the marginal alimentary lack that still persists, there are no adequate arguments for L-thyroxine monotherapy of uncomplicated iodine-deficiency goiter. PMID- 15532415 TI - [You have a child with an unusual cough? Put your money on asthma]. PMID- 15532416 TI - [Acute emesis]. AB - Causes of acute vomiting are mostly harmless and readily amenable to treatment, so that the condition thus has a good prognosis. In more rare cases, however, it might also be a symptom accompanying a more serious--even life-threatening- illness. A meticulous diagnostic investigation and effective treatment is therefore mandatory. PMID- 15532417 TI - [New EBM: is it really so detrimental to physicians?]. PMID- 15532418 TI - [Internists warn: do not force physicians into blind acceptance]. PMID- 15532419 TI - [Management of long-term anticoagulation. Narrow margin between effectiveness and risk]. PMID- 15532420 TI - [10 minute consultation: hypokalemia as incidental finding]. PMID- 15532421 TI - [Diagnosis at a glance. Difference in pupils without symptoms]. PMID- 15532422 TI - [Cardiovascular prevention: new study, new guidelines. The effort keeping the heart healthy]. PMID- 15532423 TI - [RAS blockade in diabetic nephropathy. "Early intervention can dramatically decrease loss of function]. PMID- 15532424 TI - [Increasingly more important as a source of information. Patient self-help groups: competition for the physician? (interview by Dr. Peter Kupfer)]. PMID- 15532425 TI - [Diagnosis and therapy in stomach cancer: unfortunately there are some errors!]. PMID- 15532426 TI - [Mild cognitive disorders should not be treated lightly]. PMID- 15532427 TI - ["Mild" cranio-cerebral trauma occasionally have grave psychic sequelae]. AB - Cranial injuries appear as neurological symptoms such as unconsciousness and focal neurological deficits. Mild cranio-cerebral traumas, that is, brain injuries without a serious neurotraumatologic symptom complex occuroften in practice. Nevertheless, even slight injuries, above all in the frontal cortical brain areas, can be accompanied by characteristic psycho-organic symptoms ("mild cognitive impairment" in the form of a "pseudoneurasthenic syndrome"). They are detected only after a thorough clinical psychiatric examination with a concomitant neuropsychological test. The diagnostics should be carried out as earlyas possible in order to identify and to immediately treat the respective disease. Chronicity and development of a depressive disorder can be, thus, prevented. PMID- 15532428 TI - [Mild cognitive impairment]. AB - From today's point of view, patients with a mild cognitive impairment (MCI) are at risk for developing a dementia. This clinical picture occurs frequently in the population and is easily recognized through simple tests and interviews conducted in the general and neurological medical practice. A goal of the diagnostics is to detect potentially redressable causes, above all depressive disorders, and to introduce a therapy. In many cases, however, MCI is an early stage of Alzheimer's disease or other neurodegenerative processes. When such a cause is suspected, the most important medical measure is to monitor the progression of the disease so that when deterioration occurs, therapy with antidementia drugs can promptly begin. In the future, patients with MCI will play an increasingly important role as the target group for the prevention of dementia. PMID- 15532429 TI - [Neurochemical early and differential diagnostics for Alzheimer's disease]. AB - Current medication procedures for Alzheimer's dementia (AD) such as inhibitors of acetyl cholinesterase or Memantine and future therapeutic approaches demand improved early diagnostics. However, the differentiation between early cognitive impairments due to primary progressive demential conditions and cognitive deficits in geriatric or depressive people is a great clinical challenge. The detection of biochemical markers such as beta-amyloid peptide and tau proteins in the cerebrospinal fluid can lead to improved diagnostics of the early stages and premonitory symptoms of AD. As a result, future preventive drug treatment strategies can be employed early and selectively. PMID- 15532430 TI - [Differential diagnosis and treatment of chronic cough]. AB - Postnasal drip, asthma and gastroesophageal reflux disease are the underlying causes in almost 90% of cases with chronic cough. Causal treatment is successful in the majority of patients, although in the event of a long-standing cough, it might need to be continued over several weeks. Smoking complicates the identification of cough as a clinical early symptom of an underlying tumor. Cardiac causes are rare, and in most cases are due to the use of ACE-inhibitors. Cough may be triggered by a variety of causes and the therapeutic palette must include several spectra. Since the individual causes often cannot be unequivocally identified, it may be necessary to take a polypragmatic therapeutic approach targeting the three most common causes simultaneously for 7-10 days. PMID- 15532431 TI - [One hypertensive in ten affected? Renaissance of primary hyperaldosteronism]. PMID- 15532432 TI - [First country-wide family physician model: 1 euro per patient and month]. PMID- 15532433 TI - [As a result of unpaid general practice fee: first health insurance complaint against patients]. PMID- 15532434 TI - [Diagnosis at a glance: leg ulcers and necroses after urinary tract infection]. PMID- 15532435 TI - Successful treatment of hydromorphone-induced neurotoxicity and hyperalgesia. AB - There has been an increase in opioid consumption world wide in the last decade. There has also been a disturbing increase in the number of reports of neuroexcitatory opioid-related side effects observed in patients receiving large doses of systemically administered morphine and its structural analogue, hydromorphone. It is now becoming clearer that patients receiving long-term opioid therapy can develop unexpected pain. We describe an interesting case of successful management of hydromorphone-induced neurotoxicity and hyperalgesia produced by short-term therapy with rapidly escalating doses of systemic hydromorphone. PMID- 15532436 TI - Thrombolytic therapy in stroke-related trauma. AB - PURPOSE: Trauma and/or evidence of nonintracranial bleeding is a relative contraindication for thrombolytic therapy in acute stroke. The aim of this study was to determine the incidence and nature of stroke-related trauma and its influence on treatment of patients. METHODS: Patients with stroke-related trauma were identified among all acute-stroke patients protocoled for intravenous/intraarterial thrombolytic therapy during a two-year period (6/01 5/03). The history and physical examination along with imaging findings and hospital records of these patients were reviewed. RESULTS: Six patients with stroke-related trauma were identified among 520 protocoled patients. In three, trauma resulted from a fall due to stroke and in three, trauma resulted from a motor vehicle crash due to stroke. Two from the former group sustained fracture (one each wrist and hip) and one from the latter group sustained fracture (sternum) with soft-tissue injury alone in the remaining three. Three of the six stroke-related trauma patients were excluded from consideration of thrombolytic therapy. CONCLUSIONS: Stroke-related trauma occurred equally from fall and motor vehicle crash in six of 520 patients protocoled for thrombolytic therapy, three with soft-tissue injury alone and three soft-tissue injury and fracture. Three of the six were inappropriately excluded from consideration of thrombolytic therapy. PMID- 15532437 TI - Accelerated idioventricular rhythm in a healthy teenager post circumcision. AB - Accelerated idioventricular rhythm (AIVR) is a ventricular arrhythmia, most commonly seen in adults with underlying cardiac disease. Few cases of AIVR have been reported in the adult population in the absence of cardiac pathology. We describe a case of a healthy teenager who developed episodes of AIVR postoperatively. Extensive evaluation with echocardiogram, cardiac MRI, exercise stress test, and Holter monitoring did not reveal any gross abnormalities except for mitral valve prolapse. Our patient was asymptomatic at all times and did not have any serious cardiac problems. PMID- 15532438 TI - Macrolide resistance in Streptococcus pneumoniae: mechanisms, patterns, and clinical implications of resistance. PMID- 15532439 TI - Are physicians really listening to their patients? PMID- 15532440 TI - Destiny--more than Geography! PMID- 15532441 TI - Technology and bioethics: two interviews and a forum. PMID- 15532442 TI - Termination. PMID- 15532443 TI - More "on chriopractic". PMID- 15532444 TI - [High tech: an open and permanent confrontation of ideas in interventional cardiology]. PMID- 15532445 TI - [How to assess coronary lesion, reperfusion quality, and PCI result?]. AB - During High Tech congress 2004, different questions have been discussed concerning the assessment of cardiac ischemia, quality of reperfusion and result of angioplasty. Coronary angiogram alone is not enough to decide the optimal therapeutic strategy. Ischemia-driven decision is a much better option. How should we assess the more accurately the need for ischemia-driven revascularization? Angiographic success (TIMI 3 flow grade) after angioplasty is near 95%, but is not sufficient to assess precisely the quality of myocardial reperfusion. Which more efficient tools are available? At last, which angiographic or clinical criteria should we use to assess the result of angioplasty? PMID- 15532446 TI - [Drug eluting stents: lessons from registries and literature]. AB - Discussion about drug eluting stents (DES) was a highlight of the High Tech 2004 congress. This new kind of coronary stent represents a new revolution in the field of coronary angioplasty. After the excellent results with the bare metal stent compared with balloon angioplasty, concerning clinical events and restenosis in the 90s, DES have showed a spectacular decrease in restenosis rate. If it does not mean the end of restenosis, its decrease opens new therapeutic perspectives in percutaneous coronary intervention. A lot of registries and trials are now available and allow to assess the current use of DES, assess the incidence of (acute) stent thrombosis, assess the new therapeutic indications (intra-stent restenosis, left main coronary artery, chronic total occlusion, acute myocardial infarction...) and experimentations concerning evolution of the device are in process. PMID- 15532447 TI - [Percutaneous correction of acquired aortic stenosis and mitral insufficiency in adults]. AB - Aortic stenosis and mitral regurgitation are the most common valvular diseases in western countries. Surgical treatment, aortic valve replacement or mitral valve repair, is the treatment of choice. Innovative technologies could offer an alternative therapeutic option to the patients with increased operative risk. This article will report the early experience with these promising techniques as presented at the High-Tech 2004 meeting. PMID- 15532448 TI - [Non-invasive coronary angiography by multidetector spiral computed tomography and magnetic resonance imaging]. AB - Coronary angiography is the reference standard to confirm the presence and severity of coronary stenoses. Given the invasiveness of this procedure, a noninvasive mean allowing to visualize coronary anatomy would be of obvious clinical interest. Multidetector spiral computed tomography and magnetic resonance imaging are new and promising techniques for noninvasive detection of significant coronary stenoses. The purpose of this article is to review the present knowledge on these noninvasive techniques. PMID- 15532449 TI - [The use of the RESCUE thromboaspiration system in acute coronary syndrome]. AB - Interventional procedures associated with acute coronary syndromes or performed on saphenous bypass grafts frequently lead to embolic complications, resulting in no-reflow phenomenon, side-branch occlusion, or peri-procedural infarction. The RESCUE thrombo-aspiration system was used in 19 percutaneous coronary interventions. After initial use of the aspiration device, 81% of procedures were followed by stent deployment. TIMI flow 2 or higher was present in 42% at the beginning of the procedure and in 95% at the end. In-hospital MACE rate was 4.76%. This relatively user-friendly technique appears rapid and efficacious in the case of visible intracoronary thrombus. PMID- 15532450 TI - [Prevention of recurrences of atrial fibrillation by cardiologists established in general office practice: data from the FAUVE observatory]. AB - OBJECTIVES: Analyse the modalities of preventive therapy of recurrences of paroxysmal or persistent atrial fibrillation (AF) with Vaughan-Williams (VW) type IC antiarrhythmics. METHODS: Observational study conducted with 326 French cardiologists established in general office practice, involving on the one hand an opinion survey among the cardiologists and on the other hand a cross-sectional observatory of usual medical practice. Each cardiologist was asked to include two patients aged less than 65 with non-permanent (paroxysmal or persistent) AF without left ventricle dysfunction (LVD) and initiated on treatment with a VW type IC antiarrhythmic after cardioversion to sinus rhythm. RESULTS: The opinion survey among the cardiologists indicates that non-permanent AF constitutes 36.1% of AF cases, of which 57.8% concern LVD-free patients. Most cardiologists (85%) declare to institute a preventive therapy of AF recurrences in 70-100% of these patients after cardioversion to sinus rhythm, with a VW type IC antiarrhythmic in more than 50% of cases. Of the 633 patients included in the FAUVE observatory, mainly men, 409 (64.6%) had paroxysmal AF and 224 (35.4%) had persistent AF. Analysis of therapeutic management shows that both alteration of the previous treatment and the choice of a VW type IC antiarrhythmic are based chiefly on efficacy and on tolerability of the antiarrhythmic therapy. CONCLUSION: VW type IC antiarrhythmics constitute a therapy of choice for the maintenance of sinus rhythm in non-aged and LVD-free patients with non-permanent AF. PMID- 15532451 TI - [Calpains and cardiac diseases]. AB - Calpains are a large family of cytosolic cysteine proteases composed of at least fourteen distinct isoforms. The family can be divided into two groups on the basis of distribution: ubiquitous and tissue-specific. Our current knowledge about calpains properties apply mainly to the ubiquitous isozymes, micro- and milli-calpain (classic calpains). These forms are activated after autolysis. Translocation and subsequent interactions with phospholipids of these enzymes increase their activity. Calpains are able to cleave a subset of substrates, as enzymes, structural and signalling proteins. Cardiac pathologies, such as heart failure, atrial fibrillation or clinical states particularly ischemia reperfusion, are associated with an increase of cytosolic calcium and in this regards, calpain activation has been evoked as one of the mediators leading to myocardial damage. Calpain activities have been shown to be increased in hearts experimentally subjected to ischemia reperfusion or during hypertrophy, but also in atrial tissue harvested from patients suffering from atrial fibrillations. These activities have been related to an increase of the proteolysis of different myocardial components, particularly, troponins, which are major regulators of the contraction of cardiomyocytes. Moreover, recent works have demonstrated that calpains are involved in the development of myocardial cell death by necrosis or apoptosis. PMID- 15532452 TI - [Does symptomatic stable coronary artery disease still exist in France?]. AB - In spite of the continuous growth of myocardial revascularisation techniques, stable angina remains an important clinical burden. In France, and based upon estimates from the European Society of Cardiology and from diverse epidemiological sources, 2 million people are likely to have stable angina. Though primary and secondary prevention are improving, this figure is likely to increase further, in particular because of the constant aging of the population (20% of patients more than 80 years of age have angina), but also because of the epidemics proportions that diabetes mellitus and obesity take. PMID- 15532453 TI - [Stable coronary artery disease: have we failed?]. PMID- 15532454 TI - [Acute coronary syndromes and pheochromocytoma]. AB - Pheochromocytoma can induce a adrenergic cardiomyopathy. This one occurs sometimes clinical, biological or and electrocardiographic signs of an acute coronary syndrome. We report two cases of rudimentary necrosis due to a pheochromocytoma: the first one after tumorectomy, the second one has revealed the disease in a patient who was suffering from high blood pressure for many years. The coronarography was in each case normal. PMID- 15532455 TI - [Traumatic dissection of the circumflex coronary artery complicating a myocardial contusion. A case report]. AB - Myocardial contusion can result in lethal complications as myocardial infarction, tamponade and acute valvular lesions. Following blunt thoracic trauma, the diagnosis is often difficult to make because of the multiple associated injuries. We report the case of a 42-year-old woman who, after a blunt chest trauma, developed a tamponade, an acute transmural myocardial infarction and an acute mitral regurgitation. Coronary arteriography revealed a dissection in the circumflex artery. PMID- 15532456 TI - [Influence of the sociodemographic and clinical profile of dyslipidaemic patients on their compliance with dietary advice combined with cholesterol-lowering drug treatment (nutrivast survey)]. AB - OBJECTIVE: To describe the compliance of dyslipidaemic patients with health and dietary advice combined with cholesterol-lowering drug treatment according to their sociodemographic and clinical profile. MATERIAL AND METHOD: French cross sectional epidemiological survey conducted in dyslipidaemic patients requiring introduction of lipid-lowering treatment, based on a combination of clinical description of the patient by the doctor and a strictly anonymous self administered questionnaire returned directly to the data analysis centre, allowing patients to freely express themselves about their compliance with health and dietary advice. RESULTS: 7902 patients with a mean age of 58 +/- 12 years, predominantly males (62.3%) were included in the study. The study of eating behaviour showed that 24.9% of patients presented poor compliance with dietary advice. Compliance was better in women, in patients over the age of 60, in people living at home or retired than in working or unemployed people, in urban dwellers than in rural dwellers, in patients eating lunch and dinner at home rather than in a restaurant. Patients with the greatest number of cardiovascular risk factors presented the poorest compliance with dietary advice, patients in a secondary prevention setting do not present better compliance than patients in a primary prevention setting, and those whose previous treatment was insufficient do not comply with dietary advice better than those in a treatment initiation setting. CONCLUSION: This study shows that the fundamental value of diet appears to be poorly understood by patients, who perceive it exclusively as a way of decreasing their cholesterol level and not as a measure able to improve their global cardiovascular prognosis. This probably explains why so many patients no longer see the point of diet once they have started lipid-lowering treatment. PMID- 15532457 TI - [Mycobacterial infections: the past, present, and future]. PMID- 15532458 TI - [The specific features of tuberculosis in the biogeochemical subregions of the Republic of Chuvashia]. AB - The specific features of the health status of the indigenous population, the risk factors of tuberculosis, which determine the different rates of mortality, the differences in the clinical picture of complications, concurrent diseases, the causes of death in relation to the biogeochemical conditions of the environment have been traced in the Republic of Chuvashia. With this, the differences are of great value in the content of trace elements and their ratios. The similar features in the basic epidemiological parameters have been found in the biogeochemically specific Aldan and Abyi districts of the Republic of Sakha (Yakutia). A long-term chronic experiment on inbred CBA mice has shown the value of the deficiency and excess of the trace element silicon in the formation of resistance to tuberculous infection. The findings have predetermined the feasibility of using the silicon preparation silicea in the combined treatment of patients with pulmonary tuberculosis with antituberculous agents that are highly effective and inexpensive. PMID- 15532459 TI - [Group IIIA Dispensary Registration Group among children and adolescents and its epidemiological significance]. AB - In recent years, the detection rate of signs of spontaneous tuberculosis cure is 4.1 per 100,000 children and 2.2 per 100,000 adolescents. Those who are ill in a risk group supplement the total number of new cases among children and adolescents only by 0.4 and 0.05%, respectively. The use of currently radiation diagnostic techniques, including computed tomography, reduces the number of residual changes of tuberculous etiology. PMID- 15532460 TI - [Pathomorphological assessment of the course of tuberculosis in Tashkent]. AB - Fatal cases of tuberculosis were analyzed in 3 groups: 1) Tashkent citizens registered in tuberculosis dispensaries (83.39%); 2) those unregistered there (11.1%); 3) Tashkent migrants (5.6%). Main clinical entities of tuberculosis, such as fibrocavenous tuberculosis (88.9%), infiltrative tuberculosis (3.7%), and disseminated tuberculosis (3.7%) were found to be causes of death. PMID- 15532461 TI - [The epidemic situation of tuberculosis in the Tver Region]. AB - Mathematical analysis of the basic parameters was used to trace a trend in the epidemiological situation of tuberculosis in the Tver Region. Unfavorable shifts were noted in the increase of the prevalence, incidence, and mortality of tuberculosis among children and adolescents, and a rise in the reservoir of tuberculous infection. New approaches to improving the epidemiological situation by integrating the phthisiological service and general health care facilities, by enhancing a role of a family physician, by applying informational systems for monitoring the epidemic situation, and by introducing the adapted international strategies, WHO recommendations have been developed for the treatment of patients and tested. It is suggested that the tuberculosis-controlling service should closely interact with governmental bodies, social and religious organizations, and the mass media for the optimization of tuberculosis control at a regional level. PMID- 15532462 TI - [Fibrocavernous pulmonary tuberculosis in the Sverdlovsk Region]. AB - Among new cases of fibrocavernous pulmonary tuberculosis and those who have died from it, the proportion of females and subjects with postmortem diagnosis increases and that of prisoners and the accused decreases. The institutions of confinement do not greatly affect the general regional morbidity and mortality due to fibrocavernous pulmonary tuberculosis. The new cases of fibrocavernous pulmonaary tuberculosis and those had died from it were mainly males aged 45 years or older who had a very low social status, lived in large towns and died outside a hospital, which substantiated forensic medical autopsy. Most of them had not consulted a doctor or long followed up for tuberculosis. This led to the fact that postmortem diagnosis was made in 62.6% of the cases. PMID- 15532463 TI - [Intensive chemotherapy tolerance in old patients with pulmonary tuberculosis]. AB - In accordance with the WHO recommendations, the intensive stage of chemotherapy was performed in 100 patients aged 60 to 87 years who had pulmonary tuberculosis. Most (n = 78) patients successfully completed this stage and the remaining 22 patients developed intractable adverse reactions and they had to receive individual chemotherapy regimens. In addition, 27 more patients had adverse reactions that could be eliminated by routine methods, without discontinuing standard treatment. The distinctive feature of intractable reactions were their occurrence within the first 10-20 days after the onset of treatment, they were markedly toxic and allergic and appeared as changes in some organ systems. They are most likely to develop in patients with significant cardiovascular diseases (hypertensive disease, coronary heart disease) accompanied by pronounced focal and diffuse myocardial changes, as evidenced by ECG and in those with signs of prior myocardial infarction. Correctable adverse reactions generally occur in the final period of intensive chemotherapy, they are mainly toxic and differ from intractable reactions of less extent. PMID- 15532464 TI - [Psychoneurotic disorders in ineffectively treated patients with first diagnosed destructive pulmonary tuberculosis]. AB - The evolution and specific features of psychoneurotic symptoms were studied in 270 patients with pulmonary tuberculosis. Four stages of evolution of psychoneurotic disorders were identified. Their pattern and syndromes were described. Their diagnosis and correction were recommended for patients with tuberculosis. PMID- 15532465 TI - [Surgical treatment of obstructive urinary tract diseases in patients with pulmonary tuberculosis]. AB - The paper presents the data of an examination of 92 patients [males (59-64.1%) and females (33-35.9%)] with active pulmonary tuberculosis concurrent with obstructive urinary tract diseases of various etiology. The patients' age ranged from 39 to 90 years (mean 58.2 +/- 2.5 years). Seventy-six (82.0%) patients suffered from infiltrative tuberculosis; 12 (13.0%) and 4 (4.3%) patients had cavernous and fibrocavernous tuberculosis, respectively. A relationship was found between pulmonary tuberculosis and obstructive urinary tract diseases leading to uniform changes in the upper urinary tract. The use of currently available methods for examination and treatment to eliminate infra- and supravesical obstruction permits performance of surgical interventions in patients with active pulmonary tuberculosis. PMID- 15532466 TI - [Altered parameters of respiration mechanics as compared with clinical and X-ray changes in new cases of pulmonary tuberculosis]. AB - This study was undertaken to explore changes in the mechanics of respiration in new cases of pulmonary tuberculosis during 3-4-month chemotherapy as compared to the course of a tuberculous process, which was based on the times of bacterial isolation cession and cavernous closures. A hundred and thirty-four patients (95 males and 39 females) with different forms of tuberculosis were followed up. The study of external respiratory function involved spirometry, determination of forced expiratory flow-volume, body plethysmography, examination of the mechanics of the lung by esophageal probing. During chemotherapy, bronchial patiency, the elastic constituent of pulmonary impedance, total bronchial resistance, and respiratory performance were ascertained to vary independently of one another. The authors have identified the factors that most greatly influence these changes: baseline bronchial obstruction on the one hand and the form of tuberculosis and the severity of a destructive process in the lung on the other one. A significant association of the time course of changes was found in the mechanics of the lung and in a tuberculous process, which was based on the times of bacterial isolation cession and cavernous closures. PMID- 15532467 TI - [The level of lipid peroxidation products and the parameters of antioxidative defense in the expired air condensate in patients with respiratory tuberculosis]. AB - The paper presents the data of examination of three groups of patients with pulmonary tuberculosis: 25 with a relatively favorable course of the disease, 26 with acutely progressive tuberculosis, pneumoperitoneum along with chemotherapy was used in complex therapy in 10 of them. The parameters of lipid peroxidation were estimated in the condensate of the expired air from patients before and 3 months after inpatient therapy. Irrespective of the type of the course of the disease, patients with tuberculosis were found to have a marked increase in the elimination of lipid peroxidation products, as well as deficiency of antiradical defense factors, which also preserved after therapy. The use of pneumoperitoneum in the complex therapy of patients with acutely progressive tuberculosis reduced the level of lipid peroxidation products more significantly. PMID- 15532468 TI - [Antimicrobial activity of blood phagocytes in patients with tuberculosis and respiratory sarcoidosis]. AB - Examination of 82 patients has revealed that pulmonary tuberculosis is characterized by a considerable increase in the antimicrobial activity of phagocytes: the drastic elevation of the plasma level of neutrophilic elastase and the changes opposite in direction in the phagocytic content of nonenzymatic cation proteins and myeloperoxidase. There is evidence for that the activation of circulating phagocytes is much less pronounced in patients with respiratory sarcoidosis. The significantly less plasma activity of neutrophilic elastase and the significantly greater phagocytic values of cation proteins suggest that the synthesis of secretion products prevails over their degranulation in sarcoidosis. It has been shown that the simultaneous determination of nonenzymatic cation proteins and myeloperoxidase in the phagocytes of patients allows the diagnosis to be established in 33.3% of cases. PMID- 15532469 TI - [Validity of the causes of death from tuberculosis as evidenced by medical death certificates]. AB - Despite the violations made in filling in "Medical Death Certificates", the rate of tuberculosis mortality forms with a high degree of significance. To minimize the above pitfalls, it is necessary to keep a computer-aided record of deaths from tuberculosis, which involves the standardization of entries of a postmortem diagnosis, by decoding it in accordance with the lines of item 1 (a, b, c) of the "Certificate". PMID- 15532470 TI - [Sarcoidosis and pregnancy: data from scientific publications and a clinical case]. AB - The paper analyzes the literature on sarcoidosis and pregnancy, available from the MedLine bases and Russian journals. It also presents a clinical case of sarcoidosis developed and recurred in a female after the first and repeated pregnancy. It is concluded that pregnancy in sarcoidosis may be preserved in most cases. Life-threatening rhythm and conduction disturbances, significant failure of the organs and systems afflicted by sarcoidosis, as well as female genital sarcoidosis may be a reason for deciding whether pregnancy should be preserved. PMID- 15532471 TI - [Alcohol sales and mortality due to pulmonary tuberculosis: relationships at a populational level]. PMID- 15532472 TI - [Tuberculosis in the tsar's family: Cesarevitch Georgi Aleksandrovich]. PMID- 15532474 TI - [Current problems in the performance and improvement of sociohygienic monitoring]. PMID- 15532475 TI - [Research problems in the improvement of sociohygienic monitoring]. PMID- 15532476 TI - [Experience with sociohygienic monitoring in Russia]. PMID- 15532477 TI - [A place and a role of radiation hygienic monitoring in the sociohygienic monitoring system]. PMID- 15532478 TI - [Sociohygienic monitoring of residential space]. PMID- 15532479 TI - [Methodology for assessing the quality of soil for sociohygienic monitoring]. PMID- 15532480 TI - [Epidemiological surveillance of nosocomial infections and the sociohygienic monitoring system]. PMID- 15532481 TI - [Sanitary-and-epidemiological safety and space ecological-and-hygienic zoning]. PMID- 15532482 TI - [Experience in setting up an information analytical base of sociohygienic monitoring in the system of the Ministry of Justice of Russia]. PMID- 15532483 TI - [Assessment of ambient air quality in the localities by the calculation method in the sociohygienic monitoring]. PMID- 15532484 TI - [Choice of priority substances in the drinking water and evaluation of their combined effects]. PMID- 15532485 TI - [Methodology for developing a sociohygienic monitoring subsystem in preschool educational institutions]. PMID- 15532486 TI - [Problems in the chemoanalytical provision of sociohygienic monitoring]. PMID- 15532487 TI - [Biological monitoring is a constituent of sociohygienic monitoring]. PMID- 15532488 TI - [Study of immunity in the population during sociohygienic monitoring]. PMID- 15532489 TI - [The assortment index of pesticide load of areas in the sociohygienic monitoring system]. PMID- 15532490 TI - [Territorial analysis in the sociohygienic monitoring system]. PMID- 15532491 TI - [Experience with sociohygienic monitoring in Moscow]. PMID- 15532492 TI - [The specific features of organization of sociohygienic monitoring under the conditions of a large industrial town]. PMID- 15532493 TI - [Sociohygienic monitoring in the Northern Administrative District of Moscow]. PMID- 15532494 TI - [Hygienic evaluation of the human environment in Rostov-on-Don]. PMID- 15532495 TI - [The specific features of organization of sociohygienic monitoring in the Republic of Mariy El]. PMID- 15532496 TI - [Major environmental factors that influence human health in the Novosibirsk Region]. PMID- 15532497 TI - [The specific features of sociohygienic monitoring in an area of liquidation of former chemical weapons production objects]. PMID- 15532498 TI - [Topical aspects of ecological hygienic evaluation of medical industry enterprises]. PMID- 15532499 TI - [Ambient air monitoring on the territory of oil-producing areas in the Republic of Tatarstan]. PMID- 15532500 TI - [Regional features of sociohygienic monitoring in regions of petroleum chemistry]. PMID- 15532501 TI - [The system of accreditation of risk-assessing agencies in the Russian Federation]. PMID- 15532502 TI - [Risk assessment and environmental and epidemiological studies as interrelated tools of sociohygienic monitoring at the local and regional levels]. PMID- 15532503 TI - [Assessing the risk from the effects of ambient air pollution on health in the sociohygienic monitoring system]. PMID- 15532504 TI - [Human health risk factors of the intradwelling environment]. PMID- 15532505 TI - [Assessment of carcinogenic risk for the population in the areas in vicinity of large radiation objects]. PMID- 15532506 TI - [The specific features of cancer morbidity in the Republic of Khakasia]. PMID- 15532507 TI - [Assessment of human health risk in the substantiation of managerial town planning decisions]. PMID- 15532508 TI - [Evaluation of the effectiveness of human health risk-reducing measures in poor environmental areas]. PMID- 15532509 TI - [Plenary Session of the Human Ecology and Environmental Hygiene Research Board of the Russian Academy of Medical Sciences and the Ministry of Health of the Russian Federation]. PMID- 15532510 TI - [Decision of the Plenary Session of the Human Ecology and Environmental Hygiene Research Board of the Russian Academy of Medical Sciences and the Ministry of Health of the Russian Federation on Sociohygienic Monitoring: Methodology, Regional Features and Managerial Decisions (Moscow, December 17-19, 2003]. PMID- 15532511 TI - [International Congress on Health, Education, Up-Bringing of Children and Young People in the 21st Century]. PMID- 15532513 TI - The measure of quality. AB - Reflecting an exploding interest, in performance measurement, various state, federal, and commercial entities are calling for public reporting on hospital quality. A number of measures are currently being developed or tested. Lacking, however, is a consensus about the properform and function of health care quality measures. This essay is a contribution to the growing dialog on the utility, practicality, and target audience of various measures. All measures have a subject (what they measure) and a function (the use or purpose of the measures). Subject measures shed light on process (activities prior to, and during care) or outcome (the intended or unintended result of care). Measures also fill 2 different functions: Quality improvement measures are more detailed and are designed for continuous internal consumption. Accountability measures ("report cards") are designed for external consumption, are typically shorter, less frequently generated, and reported at the hospital or clinic level. It is important that the 2 functions not be confused in constructing and targeting future quality measures. PMID- 15532512 TI - [From school hygiene to the hygiene of children and adolescents (on the occasion of the 100th anniversary of the First International Congress on School Hygiene]. PMID- 15532514 TI - Measuring performance improvement: total organizational commitment or clinical specialization. AB - BACKGROUND: Resources for hospitals are limited when they are faced with multiple publicly reported performance measures as tools to assess quality. The leadership in these organizations may choose to focus on 1 or 2 of these outcomes. An alternative approach is that the leadership may commit resources or create conditions that result in improved quality over a broad range of measures. METHODS: We used aggregated data on mortality, length of stay, and obstetrical outcomes from Greater Cleveland Health Quality Choice data to test these theories. We used Pearson correlation analysis to determine of outcomes were correlated with one another. We used repeated-measures ANOVA to determine if an association existed between outcome and time and outcome and hospital. RESULTS: All of the outcomes across all hospitals demonstrate a trend of overall improvement. Both the Pearson and ANOVA result support the hypothesis for the organization-wide approach to quality improvement. CONCLUSIONS: Hospital that make improvements in one clinical area trend to make improvements in others. Hospitals that produce improvements in limited clinical or administrative areas may not have completely adopted CQI into their culture or may not have yet realized the benefits of their organizational commitments, but use some of the concepts to improve quality outcomes. PMID- 15532515 TI - Tukey's control chart. AB - This article presents the idea of Tukey's Control Chart, a method of analyzing data based on the concepts developed by John Tukey for calculation of confidence intervals for medians. The procedure is simple to implement (no need to calculate averages or standard deviations); it does not assume any distribution of the data; it can be applied to small data sets; and it is robust and not affected by occasional unusual observations (outliers). The article provides examples of the application of Tukey's Control Chart to both patients' lifestyle management and business process improvement. PMID- 15532516 TI - Inpatients' ratings of care in 5 common clinical conditions. AB - OBJECTIVE: Using the dimensions of patient-centered care defined by the Institute of Medicine, this study examined patient satisfaction within 5 clinical conditions. METHOD: Items from the Press Ganey Inpatient Questionnaire were mapped onto the Institute of Medicine dimensions of patient-centered care. A standardized mail-out/mail-back methodology was used to collect data from random samples of patients within 5 days of discharge. SAMPLE: Patients hospitalized for heart attack, heart failure, stroke, pneumonia, or childbirth were included in the study. The sample contained 10,000 patients at 210 hospitals. RESULTS: Patients hospitalized for different clinical conditions expressed different levels of satisfaction. There may be different care needs, expectations, and evaluations of care based on these clinical conditions. However, it is probable that an all-encompassing patient-centered focus would improve care for all of these groups. CONCLUSIONS: Quality improvement professionals should attend to 2 Institute of Medicine dimensions: (1) respect for patient's values, preferences, and expressed needs and (2) emotional support, relieving fear and anxiety. It is in these areas that improved performance will be associated with the greatest increases in patient satisfaction. Four specific issues were identified as quality improvement priorities, regardless of condition: response to complaints; sensitivity to the inconvenience of hospitalization; including patients in decision making; addressing emotional and spiritual needs. PMID- 15532517 TI - Assessment of patient and provider satisfaction scales for project access. AB - OBJECTIVE: This study examined the underlying variables that contribute to patient and provider satisfaction with Project Access, a physician-driven program that connects low-income, uninsured adults (aged 18-64) to denoted specialty care and hospital services. SUBJECTS AND METHODS: Of the 550 physicians and 1400 patients participating per year, 125 physicians and 164 patients completed and returned the 14- and 15-item satisfaction questionnaires, respectively. The data from both surveys were factor analyzed. RESULTS: Patient satisfaction data factored into 4 dimensions: respect from program implementation staff, respect from pre-Project Access enrollment staff, practical health-related issues, and the level of understanding of Project Access guidelines and expectations. Provider satisfaction data factored into 3 dimensions: external services available to patients, receiving recognition and respect, and the administration of Project Access. CONCLUSIONS: Patients' feelings of respect seemed to be closely associated with their satisfaction with Project Access, in addition to the helpfulness of the program. Providers also considered respect and recognition an important factor contributing to their satisfaction, in addition to ease of administrative duties and services available to patients. PMID- 15532518 TI - A conceptual idea to improve access in a complex health care system. AB - OBJECTIVE: The purpose of this article was to briefly describe the Open Access model, a method of improving access to a health care organization, and a conceptual idea is proposed for complex organizations that want to engage in this improvement work. The target audience is health care organizations that want to improve access to their services but are challenged by the complexity of their processes. METHODS: A case study presents an organization's experience with the Open Access assessment process. This process starts with quantifying an organization's supply and demand and then deciding how to reshape its capacity and implement other strategies. A high-leverage area to improve access is a standardized or predictable process that is streamlined or implemented for a specific group of patients. RESULTS: Health care organizations are complex and have processes of care that are not explicit, in which services are rendered uniquely for each individual patient and it is difficult to see the harmony or patterns in how the work is done. CONCLUSION: The underlying principles of the Open Access model can be adopted in a complex organization, if it is acceptable to do more exploratory work and create or increase opportunities for the right things to happen. PMID- 15532519 TI - The structure, processes, and outcomes of Banner Health's corporate-wide strategy to improve health care quality. AB - Banner Health consists of 19 hospitals, 6 long-term care centers and a number of family health clinics, home care programs, and home medical equipment providers in 9 Western and Midwestern states. Banner Health has developed an integrated organization-wide effort called Care Management to simultaneously address quality and safety, reduce patient errors, and measure and report performance, outcomes, and patient satisfaction, while controlling costs through utilization management, care coordination, and performance improvement. Eleven functional areas were identified and more than 36 cross-functional and cross-facility work groups have been created. These work groups use a deliberate process in which knowledge is created, reviewed, synthesized, distributed, taught, and implemented within the system. Key lessons after the first 2 years of this effort are as follows: information sharing and collegial support can be established within newly merged organizations; there must be continued enhancement of both the accuracy and timeliness of data; the ability of health care professionals to understand and use sophisticated statistical tools has increased; a variety of methods should be used to distribute the knowledge products; and the strategy to have functional teams and work groups develop systemwide policies and toolkits but leave implementation to facility employees has worked relatively well. PMID- 15532521 TI - Interview with Lucretia Thornton, JD. PMID- 15532522 TI - How do you want to learn? PMID- 15532520 TI - Transactional versus transformational style of leadership--employee perception of leadership efficacy in public and private hospitals in Kuwait. AB - OBJECTIVES: To determine whether employees working for a transformational leader perceive their leader as more effective than do those working for a transactional leader. To assess whether employees in private hospitals are more likely to perceive their leaders as transformational than are employees in public hospitals. DESIGN: A questionnaire study of a random sample of employees working at 4 hospitals (private and public) in Kuwait; 266 respondents included. RESULTS: Analysis of varience and regression analysis showed that the transformational style of leadership was linked to high level of employee perception of leadership efficacy. Also, the employees in private hospitals were more likely to perceive their leaders as transformational than were employees in public hospitals. CONCLUSIONS: Hospitals in Kuwait may benefit by recruiting leaders with a transformational style, as it is directly related to quality, employee satisfaction, increased productivity, and employee perception of leadership efficacy. PMID- 15532523 TI - Professionalism: what it takes to restore patients' faith in us and in health care. PMID- 15532524 TI - Drug reps: open your arms or bar the door? PMID- 15532525 TI - Patient preferences prod purchase: patients help practice choose an electronic medical records system. PMID- 15532526 TI - Now hear this: practices must provide interpretive services for deaf and hearing impaired patients. PMID- 15532527 TI - 360-degree feedback systems: development and effectiveness. PMID- 15532528 TI - A PEO for your practice? Professional employer organizations can lighten your administrative burden. AB - Many medical practices turn to professional employer organizations (PEOs) to handle their employee benefits and many other administrative functions. PEOs can assume many risks, eliminate numerous headaches and provide a group's employees with a quality benefits package, often at a lower cost. A PEO can allow a group's management team to focus more on other aspects of the medical practice. Although they're not for everyone, PEOs are worth a look. PMID- 15532529 TI - For love of the profession. Award winners demonstrate exceptional accomplishments, leadership. AB - A balanced blend of passion, commitment and inspiration lift certain medical practice professionals above their peers. Learn what it takes to make exceptional achievements. The American College of Medical Practice Executives' Honors Program, the official awards program of MGMA, bestows awards for 2004 on four outstanding individuals and one organization. PMID- 15532530 TI - Selecting an EMR in only eight years: one group's experience. AB - Why did it take a 27-physician multispecialty medical practice eight years to select and implement an electronic medical record (EMR) system? The author describes the difficult assessment process and emphasizes the importance of organizational readiness and financial wherewithal. PMID- 15532531 TI - Show me the money. Case study: negotiating higher reimbursement from an insurance company. AB - Most medical practices simply accept managed care contracts and their corresponding fee schedules. But by negotiating, a group can often find bargaining room to reach a deal for a higher level of reimbursement. The author describes how he renegotiated a contract with a health plan to the practice's advantage. PMID- 15532532 TI - Flower power. PMID- 15532533 TI - Dimethyl sulfoxide and ebselen prevent convulsions induced by 5-aminolevulinic acid. AB - We investigated whether intrastriatal (i.s.) administration of 5-aminolevulinic acid (ALA) induces oxidative damage and whether behavioral alterations induced by i.s. administration of ALA could be affected by antioxidants. Unilateral injection of ALA (6 micromol/striatum) increased (approximately 30%) thiobarbituric acid-reactive substances (TBARS), but did not affect striatal content of total thiol groups. ALA-induced body asymmetry was not prevented by pretreatment with ascorbic acid (100 mg/kg, s.c.), dimethyl sulfoxide (DMSO, 0.5 microl/striatum, i.s.) or ebselen (10 nmol/striatum, i.s.). ALA-induced convulsions were not prevented by ascorbic acid, but were partially prevented by DMSO and completely prevented by ebselen. Ebselen completely prevented the increase of striatal TBARS induced by ALA. Results obtained suggest the involvement of reactive species in ALA-induced convulsions and may be of value in understanding the physiopathology of neurological dysfunctions associated to ALA overload. PMID- 15532534 TI - Ebselen protects Ca2+ influx blockage but does not protect glutamate uptake inhibition caused by Hg2+. AB - The goal of this study was to investigate the isolated and combined effect of ebselen and Hg2+ on calcium influx and on glutamatergic system. We examined the in vitro effects of 2 phenyl-1,2-benzisoselenazol-3(2H)-ona), (Ebselen) on 45Ca2+ influx in synaptosomes of rat at rest and during depolarization and glutamate uptake into synaptosomes. Entry of 45Ca was measured during exposure to mercury in non-depolarizing and depolarizing solutions. Ebselen abolished the inhibition of 45Ca2+ influx on non-depolarizing conditions; however, ebselen did no modify inhibition uptake of 45Ca2+ caused by Hg2+ in high K+ depolarizing medium. Ebselen did not modify glutamate uptake inhibition caused by Hg2+ in synaptosomes. These results indicate that ebselen has an in vitro protective effect against Hg2+ induced inhibition of Ca2+ influx into synaptosomes, depending on the depolarizing conditions of the assay. The effects of Hg2+ on glutamate uptake were not modified by ebselen, suggesting that its protection is dependent on the target protein considered. PMID- 15532535 TI - Protective effect of 1-methylated beta-carbolines against 3-morpholinosydnonimine induced mitochondrial damage and cell viability loss in PC12 cells. AB - The present study investigated the effect of 1-methylated beta-carbolines (harmaline, harmalol and harmine) on change in the mitochondrial membrane permeability and cell death due to reactive nitrogen species in differentiated PC12 cells. beta-Carbolines, caspase inhibitors (z-LEHD.fmk and z-DQMD.fmk) and antioxidants (N-acetylcysteine, dithiothreitol, melatonin, carboxy-PTIO and uric acid) depressed cell viability loss due to 3-morpholinosydnonimine (SIN-1) in PC12 cells. beta-Carbolines inhibited the nuclear damage, the decrease in mitochondrial transmembrane potential, the cytochrome c release, the formation of reactive oxygen species and the depletion of GSH caused by SIN-1 in PC12 cells. beta-Carbolines decreased the SIN-1-induced formations of 3-nitrotyrosine, malondialdehyde and carbonyls in PC12 cells. The results show that 1-methylated beta-carbolines attenuate SIN-1-induced mitochondrial damage. This results in the inhibition of caspase-9 and -3 and apoptotic cell death in PC12 cells by suppressing the toxic actions of reactive oxygen and nitrogen species, including the GSH depletion. PMID- 15532536 TI - Identification and characterization of a novel form of the human L-dopa decarboxylase mRNA. AB - L-Dopa decarboxylase (DDC) has been cloned from several species and was shown to undergo alternative splicing within its 5'-untranslated and coding regions. In this report, we describe a novel splice variant of DDC mRNA in human tissue, lacking exons 10-15 of the full-length transcript but including an alternative exon 10. The isolated alternative human DDC cDNA (alt-DDC) was cloned from human placenta, and was found to be of the neuronal type. Northern blot analysis indicated that the alt-DDC transcript is expressed in high levels in human kidney. Our results demonstrate the detection of a new alternative splicing event within the coding region of the human DDC mRNA, further suggesting that the single copy human DDC gene undergoes complex processing leading to the formation of multiple mRNA isoforms. PMID- 15532537 TI - Cyclooxygenase-mediated generation of free radicals during hypoxia in the cerebral cortex of newborn piglets. AB - Previous studies have demonstrated that free radicals are formed under hypoxic conditions in newborn piglet brain. To test the hypothesis that the cyclooxygenase pathway serves as a source of free radical generation during hypoxia studies were performed on 24 piglets divided into four groups. Six saline (group 3) and six indomethacin treated (group 4) were exposed to hypoxia (FiO2 0.05-0.07) for 60 min. Cerebral hypoxia was documented biochemically by determination of ATP and phosphocreatine. Fluorescent compounds and conjugated dienes were determined as indices of lipid peroxidation. Free radical formation was determined by using n-tert butyl phenyl nitrone (PBN) as a spin trap agent and measuring spin adduct formation in duplicate using a Varian E-109 spectrometer. Groups 1 and 2 (normoxic) showed no spin adduct formation. Group 3 showed a significant increase in spin adduct formation compared to normoxia (372+/-125 vs. 63+/-15, P<0.001). Hypoxic animals pretreated with indomethacin had a spin adduct level of 197+/-132 and were similar to normoxic animals. ATP/PCr levels were the same in groups 3 and 4 denoting the same degree of cerebral hypoxia in all hypoxic animals. Conjugated dienes increased significantly during hypoxia as compared to normoxia (0.142+/-0.017 vs. 0.0+/ 0.0) and were decreased insignificantly with indomethacin treatment. Fluorescent compounds were not significantly different among the four groups. Na+,K+-ATPase activity decreased during hypoxia but was not preserved in hypoxic animals pretreated with indomethacin. These data provide direct evidence of the presence of free radicals during hypoxia and the contribution of cyclooxygenase metabolism to their formation. PMID- 15532538 TI - Purine nucleoside phosphorylase activity in rat cerebrospinal fluid. AB - The sequential hydrolysis of purines is present in rat CSF and generates nucleosides as inosine and guanosine that are usual substrates for purine nucleoside phosphorylase (PNP). PNP catalyzes phosphorolysis of the purine nucleosides and deoxynucleosides releasing purine bases. Here we investigated the presence of PNP in CSF of rats using: i) a specific chromophoric analogue of nucleosides, 2-amino-6-mercapto-7-methylpurine ribonucleoside (MESG), and ii) an inhibitor of PNP activity, immucillin-H. Additionally, we performed a preliminary kinetic characterization (K(M): Henry-Michaelis-Menten constant; V: maximal velocity) for MESG and inorganic phosphate (Pi). The values of K(M) and V for MESG (n = 3, mean+/-SD) were 142.5+/-29.5 microM and 0.0102+/-0.0006 U mg(-1), respectively. For Pi (n=3, mean+/-SD), the K(M) values and V were 186.8+/-43.7 microM and 0.0104+/-0.0016 U mg(-1), respectively. The results indicated that PNP is present in rat CSF and provided a preliminary kinetic characterization. PMID- 15532539 TI - Salviae miltiorrhizae radix inhibits superoxide generation by activated rat microglias and mimics the action of amphetamine on in vitro rat striatal dopamine release. AB - Salviae miltiorrhizae Radix (SMR), an eminent herb in the treatment of cardiovascular disorder (called blood stasis in traditional Chinese medicine), is widely used in China, Japan, Taiwan and Korea. SMR is also herbal medicines used in the treatment of drug addiction without scientific support for their mechanism of action. We evaluated the effect of SMR on superoxide production by rat microglias using a 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin 3-one-dependent chemiluminescence assay. SMR dose-dependently inhibited superoxide production by microglias stimulated with phorbol myristate acetate or opsonized zymosan, while it had no effect on superoxide production by a hypoxanthinexanthine oxidase system. These results indicate that SMR does not have a scavenging effect, but has an inhibitory effect on superoxide generation by microglias. Although SMR is commonly used for treating chronic cerebral infarction, it may also have a protective effect on progression of Parkinson's disease or Alzheimer's disease. On the other hand, the present study investigated the effect of the medicinal plant on dopaminergic neurotransmission in comparison with amphetamine. The effect of crude water extracts (0.1 g/ml) of SMR on K+ (20 mM)-stimulated dopamine release from rat striatal slices was compared with amphetamine (10(-4) M) using high-performance liquid chromatography with electrochemical detection to measure endogenous dopamine. Amphetamine and SMR significantly increased K+-stimulated dopamine release (P<0.001) from rat striatal slices when compared with K+-stimulated alone. SMR potentiated the effect of amphetamine on K+-stimulated dopamine release (P<0.001) when compared with amphetamine alone. The results indicate that SMR may stimulate dopamine release in the same manner as amphetamine. It remains to be determined whether the effect of this extract on dopamine function is important in its therapeutic use in the treatment of drug addiction. PMID- 15532540 TI - The tyrosine [correction of tryrosine] phosphorylation and cytoskeletal translocation of phospholipase C gamma 1 in bovine adrenal medullary chromaffin cells. AB - The tyrosine phosphatase inhibitor BpV(phen) stimulated a concentration-dependent increase of phospholipase C (PLC) activity in bovine adrenal medullary chromaffin cells. This response was accompanied by an increase in PLCgamma1 tyrosine phosphorylation and its cytosketetal translocation. Insulin, at high concentrations, stimulated PLC activity to a similar extent as BpV(phen), a response that was also accompanied by an increase in PLCgamma1 translocation but not its tyrosine phosphorylation. BpV(phen) strongly enhanced the insulin stimulated increase in PLC activity and caused a small rise in PLCgamma1 translocation above that seen with insulin alone. Despite the synergistic rise in activity PLCgamma1 tyrosine phosphorylation did not increase beyond that seen with BpV(phen) alone. These results indicate that PLCgamma1 activation in chromaffin cells may be more closely associated with its cytoskeletal translocation than its tyrosine phosphorylation although other factors may also be important for activation of enzyme activity. PMID- 15532541 TI - Effects of metabotropic glutamate receptor stimulation on blood-brain barrier permeability during focal cerebral ischemia. AB - This investigation was performed to evaluate whether ACPD [(1S, 3R)-1 aminocyclopentane-1, 3-dicarboxylic acid], a metabotropic glutamate receptor agonist, would enhance the degree of increase in blood-brain barrier (BBB) permeability caused by focal cerebral ischemia. In this study, male Wistar rats were placed in control (n = 7) and ACPD (n = 7) groups under isoflurane anesthesia. Twenty minutes after middle cerebral artery (MCA) occlusion, patches of 10(-5) M ACPD or normal saline were placed on the ischemic cortex (IC) for a period of 40 min. Patches were changed every 10 min. One hour after MCA occlusion, BBB permeability was determined by measuring the transfer coefficient (Ki) of [alpha-14C] aminoisobutyric acid. There were no statistical differences in systemic blood pressures and heart rates between these groups. Blood gases were within normal limits. In the control group, the Ki of ischemic cortex (IC) was 2.1 times that of the contralateral cortex (CC) (3.7+/-0.9 vs. 1.8+/-0.3 microl/g/min). In the ACPD group, the Ki of the IC was 3.3 times that of the CC (5.0+/-0.7 vs. 1.5+/-0.4 microl/g/min). The increase in Ki of the ACPD group in the ischemic cortex was significantly greater than that in the control group. There was no significant difference in the Ki of the CC between these groups. Our data suggest that activation metabotropic glutamate receptors in the cortex can further augment the increase in BBB permeability caused by focal ischemia. PMID- 15532542 TI - Chaperone-like manner of human neuronal tau towards lactate dehydrogenase. AB - In our experiments, inactivation of lactate dehydrogenase (LDH, EC1.1.1.27) in the presence of human microtubule-associated tau is observably suppressed during thermal and guanidine hydrochloride (GdnHCl) denaturation. Kinetic studies show tau can prevent LDH from self-aggregation monitored by light scattering during thermal denaturation. On the other hand, neuronal tau promotes reactivation of LDH and suppresses self-aggregation of non-native LDH when GdnHCl solution is diluted. Furthermore, the reactivation yield of LDH decreases significantly with delayed addition of tau. All experiments were completed in the reducing buffer with 1 mM DTT to avoid between tau and LDH forming the covalent bonds during unfolding and refolding. Thus, Tau prevents proteins from misfolding and aggregating into insoluble, nonfunctional inclusions and assists them to refold to reach the stable native state by binding to the exposed hydrophobic patches on proteins instead of by forming or breaking covalent bonds. Additionally, tau remarkably enhances reactivation of GDH (glutamic dehydrogenase, EC 1.4.1.3), another carbohydrate metabolic enzyme, also showing a chaperone-like manner. It suggests that neuronal tau non-specifically functions a chaperone-like protein towards the enzymes of carbohydrate metabolism. PMID- 15532543 TI - Uptake of GABA and activity of GABA transaminase in blood platelets from children with absence epilepsy. AB - Childhood absence epilepsy (CAE) is a well-defined generalized epilepsy syndrome clinically characterized by frequent absence seizures. The aim of this study was to assess the activity of GABA transaminase (GABA-T) and the kinetic parameters of GABA uptake in platelets from patients with CAE. We studied 13 patients with CAE and eight sex- and age-matched controls. The mean activity of GABA-T was lower in patients with CAE than in controls (1.22+/-0.05 vs. 1.75+/-0.10 micromol/min/kg protein). The capacity of GABA uptake into the platelets was higher in patients using valproate (0.66+/-0.09 micromol/min/kg protein), but not in those using ethosuximide (0.34+/-0.05 micromol/min/kg protein), when compared to controls (0.26+/-0.06 micromol/min/kg protein). The affinity of the transporters was not altered. The observed peripheral alterations may indicate impaired function of brain GABAergic systems in children with absence epilepsy. PMID- 15532545 TI - An optimized fixation and extraction technique for high resolution of inositol phosphate signals in rodent brain. AB - Members of lower and higher inositol phosphates distinctly participate in signal transduction (1). Relatively little is known regarding possible biological functions of inositol phosphates in functionally different areas of the intact brain. A detailed study on the regional distribution of biologically important inositol phosphates may help elucidate their physiological functions in different brain regions in the regional tissue context. We now show a novel technique which allows fixation and subsequent dissection of whole rat brains into small volume elements for mapping of the whole range of inositol phosphates from Ins(1,4,5)P3 to InsP6. The method has been successfully applied to investigate regional differences of a broader spectrum of inositol phosphates in microdissected brain tissue and to construct 3D-maps of these signaling compounds. The technique can be particularly well employed to investigate regional changes in the spectrum of higher inositol phosphates and phosphoinositides upon neuronal stimulation induced by motor activity or drug treatment. PMID- 15532544 TI - Protein kinase C-mediated phosphorylation of Kv beta 2 in adult rat brain. AB - The phosphorylation of Kvbeta2 was investigated by different protein kinases. Protein kinase A catalytic subunit (PKA-CS) yielded the greatest phosphorylation of recombinant Kvbeta2 (rKvbeta2), with limited phosphorylation by protein kinase C catalytic subunit (PKC-CS) and no detectable phosphorylation by casein kinase II (CKII). Protein kinase(s) from adult rat brain lysate phosphorylated both rKvbeta2 and endogenous Kvbeta. The PKA inhibitor, PKI 6-22, fully inhibited PKA mediated phophorylation of rKvbeta2 yet showed minimal inhibition of kinase activity present in rat brain. The inhibitor Go 6983, that blocks PKCalpha, PKCbeta, PKCgamma, PKCdelta and PKCzeta activities, inhibited rKvbeta2 phosphorylation by rat brain kinases, with no inhibition by Go 6976 which blocks PKCalpha and PKCbetaI activities. Dose-response analysis of Go 6983 inhibitory activity indicates that at least two PKC isozymes account for the kinase activity present in rat brain. Thus, while PKA was the most active protein kinase to phosphorylate rKvbeta2 in vitro, Kvbeta2 phosphorylation in the rat brain is mainly mediated by PKC isozymes. PMID- 15532546 TI - Effects of lovastatin and pravastatin on amyloid processing and inflammatory response in TgCRND8 brain. AB - Previous studies suggest that treatment with statins reduce beta amyloid (Abeta) deposition in brains of mouse models of Alzheimer's disease (AD) and may reduce the prevalence of AD in humans. Since lipophilicity influences the biological efficacy of statins, we compared the effects of lovastatin, a lipophilic statin, to effects of the hydrophilic pravastatin on amyloid processing and inflammatory responses in brain. Three-month old TgCRND8 mice expressing mutant human amyloid precursor protein (mHuAPP) were treated daily with various doses of either statin. After 1 month, levels of cerebral soluble and fibrillar Abeta peptides, soluble sAPPalpha, and inflammatory cytokines were measured. Both statins caused dose-dependent reductions in total Abeta peptides with parallel increases in total sAPPalpha. At all doses, slightly greater effects were observed with lovastatin than with pravastatin. In contrast, only lovastatin significantly increased levels of IL-1beta and of TNFalpha in a dose-dependent manner. Lovastatin, but not pravastatin, decreased succinic dehydrogenase and increased lactate dehydrogenase activities in skeletal muscle and increased TUNEL staining in liver. Our data demonstrate that both statins shift the balance of APP processing from excessive beta-toward the normal alpha-cleavage while reducing the total amyloid burden in TgCRND8 brain and that lovastatin, but not pravastatin, potentiates cerebral inflammation and is associated with liver and muscle histotoxicity in these animals. These data show that pravastatin can reduce amyloid burden without potentiating inflammatory responses in brain and, therefore, may have a wider dose-range of safety than have lipophilic statins in the treatment or prevention of AD. PMID- 15532547 TI - In vitro proteolytic degradation of bovine brain calcineurin by m-calpain. AB - A major cause of neuronal dysfunction is due to altered Ca2+ regulation. An increase in Ca2+ influx can activate Ca2+-dependent enzymes including calpains, causing the proteolysis of its specific substrates. In the present study, calcineurin (CaN) was found to be proteolysed by a Ca2+-dependent cysteine protease, m-calpain. In the presence of Ca2+, the 60 kDa subunit (CaN A) was degraded to a 46 kDa immunoreactive fragment, whereas in the presence of Ca2+ /calmodulin (CaM) immunoreactive fragments of 48 and 54 kDa were observed. The beta-subunit (CaN B) was not proteolysed in either condition. The proteolysis of CaN A increased its phosphatase activity and rendered it totally CaM-independent after 10 min of proteolysis. The molecular weight of the proteolytic fragments suggested that the m-calpain cleaved CaN A in the CaN B binding domain. A CaM overlay experiment revealed that the CaM-binding site was present only in the 54 kDa fragment produced by CaN A proteolysis in the presence of Ca2+ /CaM. Thus, the increase in CaN A phosphatase activity observed in many neuronal disorders, may be due to the action of calpain. PMID- 15532548 TI - Estradiol protects against oxidative stress induced by chronic variate stress. AB - Neurochemical gender-specific effects have been observed following chronic stress. The aim of this study was to verify the effects of chronic variable stress on free radical production (evaluated by DCF test), lipoperoxidation (evaluated by TBARS levels), and total antioxidant reactivity (TAR) in three distinct structures of brain: hippocampus, cerebral cortex and hypothalamus of female rats, and to evaluate whether the replacement with estradiol in female rats exerts neuroprotection against oxidative stress. Results demonstrate that chronic stress had a structure-specific effect upon lipid peroxidation, since TBARS increased in hypothalamus homogenates of stressed animals, without alterations in the other structures analyzed. Estradiol replacement was able to counteract this effect. In hippocampus, estradiol induced a significant increase in TAR. No differences in DCF levels were observed. In conclusion, the hypothalamus is more susceptible to oxidative stress in female rats submitted to chronic variable stress, and this effect is prevented by estradiol treatment. PMID- 15532549 TI - Calcium-dependent protein kinase C activation in acutely isolated neurons during oxygen and glucose deprivation. AB - Glutamate excitotoxicity and necrotic cell death are characteristic features of ischemic neuronal injury. In the penumbral area, glutamate exposure is less pronounced and neuronal death is delayed. Recent studies suggest that delayed neuronal death is propagated by intracellular signalling pathways. Protein kinase C (PKC) activation may initiate apoptosis, but its role in ischemia is still not clear. In this study the PKC activity was investigated during non-excitotoxic ischemia in acutely dissociated rat CA1 neurons. During oxygen and glucose deprivation (OGD) the PKC activity measured with the fluorescent dye Fim-1 increased rapidly reaching a maximum of 31+/-8% (P < 0.05) after 5 min. When extracellular Ca2+ was depleted, the fluorescence intensity increased by 20+/-8% (P<0.05), but with a slower onset. In neurons treated with thapsigargin in a Ca2+ depleted solution, however, OGD did not trigger PKC activation. The results suggest that the PKC activation is mainly triggered by Ca2+ release from endogenous stores. PMID- 15532550 TI - Critical survey of quantitative proteomics in two-dimensional electrophoretic approaches. AB - The present review attempts to cover a number of methods that appeared in the last few years for performing quantitative proteome analysis. However, due to the large number of methods described for both electrophoretic and chromatographic approaches, we have limited this excursus only to conventional two-dimensional (2D) map analysis, coupling orthogonally a charge-based step (isoelectric focusing) to a size-based separation (sodium dodecyl sulfate (SDS) electrophoresis). The first and oldest method applied in 2D mapping is based on statistical analysis performed on sets of gels via powerful software packages, such as the Melanie, PDQuest, Z3 and Z4000, Phoretix and Progenesis. This method calls for separately-running a number of replicas for control and treated samples, the merging and comparing between these two sets of data being accomplished via the softwares just mentioned. Recent developments permit analyses on a single gel containing mixed samples differentially labelled and resolved by either fluorescence or isotopic means. In one approach, a set of fluorophors, called Cy3 and Cy5, are selected for differentially tagging Lys residues, via a "minimal labelling" protocol. A variant of this, adopts a newer set of fluorophors, also of the Cy3 and Cy5 type, reacting on Cys residues, via a strategy of "saturation labelling". There are at present two methods for quantitative proteomics in a 2D gel format exploiting stable isotopes: one utilizes tagging Cys residues with [2H0]/[2H3]-acrylamide; the other one, also based on a Cys reactive compound, exploits [2H0]/[2H4] 2-vinylpyridine. The latter reagent achieves 100% efficiency coupled to 100% specificity. The advantages and limitations of the various protocols are discussed. PMID- 15532551 TI - Nanoflow gradient generator for capillary high-performance liquid chromatography nanoelectrospray mass spectrometry. AB - A novel nanoflow gradient generator using a 10-port switching valve with two injection loops installed, which is referred to here as the "Asymptotic-Trace-10 Port-Valve" (AT10PV) nanoGR generator, has been applied to capillary high performance liquid chromatography (HPLC)-microelectrospray (microESI) or nanoelectrospray (nanoESI) time-of-flight mass spectrometry (TOF MS). In this study, performance of this capillary HPLC-micro/nanoESI-MS system was tested at a flow rate of 200 nl/min by using three typical peptides (angiotensins I, II, and III: 50 fmol each). The result demonstrated that this system provides reasonably good repeatability of peak retention times (R. S.D. of less than 0.5%). Sequential runs of a series of sample injections were performed in the same manner as conventional analysis at microflow rates. PMID- 15532552 TI - Heart-cutting two-dimensional electrophoresis in a single capillary. AB - The possibility of performing two-dimensional capillary electrophoresis (2D-CE) separations in a single capillary was investigated. For this purpose, a fraction stemming from the first dimension of the separation was selected and isolated in the capillary by evacuating out of the capillary the other undesirable compounds. Next, the isolated fraction was submitted to a second separation medium that was introduced in the capillary by electroosmotic flow. The second separation medium was able to reach the isolated fraction since the solutes were migrating in counter-electroosmotic mode. Since only one fraction is submitted to the second dimension of the separation, this new methodology is closer to a heart-cutting approach than to a true comprehensive 2D separation. However, it has the advantage of not requiring any special coupling device between capillaries since the two dimensions of the separation are performed in the same capillary. In this work, a simple mixture of synthetic polymers taken as model compounds was separated according to: (i) the charge density by free solution CE in the first dimension, and (ii) the molar masses by CE in the presence of an entangled polymer solution in the second dimension. The different strategies that were investigated to isolate the fraction at the end of the first dimension are described in detail. The influence of important experimental parameters (capillary diameter, applied pressure for mobilizing the solutes, diffusion coefficients of the solutes) on the performances of the two-dimensional separations were studied. A careful attention was paid to the influence of these parameters on the efficiency of the separations. The experimental results demonstrate that heart-cutting 2D electrophoretic separations can be performed in a capillary format using a single capillary. PMID- 15532553 TI - Column switching in zone electrophoresis on a chip. AB - This feasibility study deals with column switching in zone electrophoresis (ZE) separations on a column coupling (CC) chip. The column switching implemented into the ZE separations an on-chip sample clean up applicable for both the multicomponent and high salinity samples. In addition, complemented by different separation mechanisms in the coupled columns (channels), it provided benefits of two-dimensional separations. Properly timed column switching gave column-to column transfers of the analytes, characterized by 99-102% recoveries, delivered to the second separation stage on the chip the analyte containing fractions contaminated only with minimum amounts of the matrix constituents. A diffusion driven transport of the matrix constituents to the second channel of the chip (due to direct contacts of the electrolyte solutions in the bifurcation region), representing 0.1-0.2% of the loaded sample constituents, was found to accompany the sample clean up performed on the CC chip. This source of potential disturbances to the separation in the second channel, however, is not detectable in a majority of practical situations. With respect to a 900 nl volume of the sample channel on the CC chip, the electric field and isotachophoresis (ITP) stackings were employed to minimize the injection dispersion in the separations and concentrate the analytes. Here, the column switching, removing a major part of the stacker from the separation system, provided a tool effective in a control of the destacking of analytes. Highly reproducible ZE separations as attained in this work also for the chip-to-chip and equipment-to-equipment frames can be ascribed, at least in part, to suppressions of electroosmotic and hydrodynamic flows of the solutions in which the separations were performed. PMID- 15532554 TI - Adaptation of an evaporative light-scattering detector to micro and capillary liquid chromatography and response assessment. AB - A commercially available evaporative light-scattering detection (ELSD) system was adapted for micro and capillary LC. Therefore the various parameters involved in the droplet formation during the nebulization step in the ELSD system were studied. It was shown that the velocity term in the Nukiyama Tanasawa equation remains constant, leading to droplets of the same order of magnitude for narrow bore and capillary columns. Consequently, the ELSD modification was performed by decreasing the internal diameter of the effluent capillary tube in the nebulizer nozzle and by keeping its external diameter constant. Next, response curves for a conventional and the developed micro and capillary LC were compared as to investigate why a linear ELSD response is often obtained when used in micro or capillary LC. By splitting the flow rate post column, we showed that the nebulization process was not at the origin of the phenomenon. For ceramide III and tripalmitin, the response curves were found to be non-linear. However the curvature was less significant when the columns internal diameter decreased. Calculated particle size profiles for micro or capillary LC suggest that the particle entering the detection chamber are bigger than under conventional LC conditions. Last, triethylamine and formic acid were used to increase the response of the detector. The response enhancement, expected from previous studies, was established for the two lipids involved in this study. PMID- 15532555 TI - Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins. AB - The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 microm i.d. capillaries and their application for microHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 microL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 microm. The monolithic microHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed. PMID- 15532556 TI - Stabilization of phosphatidylcholine coatings in capillary electrophoresis by increase in membrane rigidity. AB - Divalent cations affect the stability and structure of phospholipid vesicles and also the binding and immobilization of proteins into phospholipid membranes. The effect of calcium, magnesium, and zinc on zwitterionic phosphatidylcholine (PC) coatings in fused silica capillaries for electrophoresis was the primary interest in this work. In addition, the effect of temperature on the coating stability was investigated by coating 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) liposomes at temperatures above and below the gel- to fluid-state transition. All coatings were performed with PC large unilamellar vesicles (LUV) in 40 mM N-(2 hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES) at pH 7.4 as basic solution. HEPES (40 mM) at pH 7.4 was used as background electrolyte (BGE) throughout the study. The stability of the coating was studied by measuring the electroosmotic flow. A molar ratio of 1:3 PC/Ca2+ or PC/Mg2+ gave the best coating stability owing to the increased rigidity of the phospholipid membrane furnished by the divalent metal ions. Better results were obtained with DPPC in the more rigid gel state than in the fluid state: the electroosmotic flow was much suppressed and the PC coating was stabilized. Coating the fused silica capillary with PC liposome-metal ion buffer solutions resulted in good electrophoretic separation of basic model proteins (pI-values 7.8-11.0). The electrophoretic results demonstrate the importance of stabilizing the phospholipid coating on fused silica capillaries, either by the addition of divalent metal ions (Ca2+, Mg2+, or Zn2+) or by working in the gel-state region of the phospholipid. PMID- 15532557 TI - Microchip electrophoresis with hydrodynamic injection and waste-removing function for quantitative analysis. AB - Quantitative analysis is problematic for microchip electrophoresis for several reasons including chip-to-chip variation, discontinuous sample re-loading, channel reconditioning, and electrokinetic injection bias. In this study, the capability for quantitative analysis on a flow-through based microchip electrophoresis, which provides continuous sample re-loading, channel washing, reconditioning and hydrodynamic injection as well as waste removing is demonstrated to be more quantifiable and more reproducible compared to manual electrokinetic injection method. Using the flow-through microchip with waste removing function, FITC-labeled estrogen or Rhodamine B could be continuously analyzed without significant changes (R.S.D. < 6.6%) in signal intensity for over 3 h, which is sufficient for a complete set of quantitative analysis. With the use of a phosphorylated kinase substrate as the model, a calibration curve for quantitative analysis of phosphopeptides were constructed and results indicate that both R2 value of the linearity and R.S.D. values of the peak intensity were around 0.9961 and 3.16%, respectively, without the use of an internal standard. These values were slightly improved to be around 0.9986 and 2.27%, respectively, with the use of a non-phosphopeptide counterpart as the internal standard. The potential of this flow-through device for the development of a kinase phosphorylation assay based on the quantitative method was also briefly discussed. PMID- 15532558 TI - Aggregation and other intermolecular interactions of biological buffers observed by capillary electrophoresis and UV photometry. AB - Electrophoretic and photometric experiments strongly indicate that monovalent anions, which arise by deprotonation of the nitrogen atom in zwitterionic Good's buffers 3-(cyclohexylamino)-2-hydroxy-1-propanesulfonic acid (CAPSO) and 3 morpholinopropanesulfonic acid (MOPS), spontaneously aggregate. Cationic migration of sanguinarine (SA) and chelerythrine (CHE) in highly alkaline 1,3 bis[tris(hydroxymethyl)methylamino]propane (Bis-Tris-propane), in which the concentration of cations of both alkaloids is negligible, may be explained by the existence of an aggregate, which contains uncharged sanguinarine or chelerythrine and one monovalent cation of Bis-Tris-propane at least. Tendency of tris(hydroxymethyl)aminomethane (Tris), bis (2 hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris) and Bis-Tris-propane cations to ion pairing with synthetic cluster borane anions and with fused silica markedly rises up with the size and charge of these cations. The drop in mobility of cluster borane compounds sometimes exceeds 50% of their mobility found at identical pH and ionic strength in buffers with sodium cation. The electroosmosis drop approached 70% if background electrolyte contained Bis-Tris-propane cations instead of sodium cations. Nitrate, taken as a model inorganic ion, and four randomly chosen organic anions interacted markedly less with Tris, Bis-Tris and Bis-Tris-propane cations than cluster borane anions. 2-(N morpholino)ethanesulfonic (MES) acid anions present in background electrolyte affect the ion pairing of Tris, Bis-Tris and Bis-Tris-propane cations with anionic analytes and, in this way influence also mobilites of these anionic analytes. Limited hydrophilicity at least one of interacting species appears to be the most probable cause of observed intermolecular interactions of biological buffers. PMID- 15532559 TI - Capillary electrophoretic studies on the migration behavior of cationic solutes and the influence of interactions of cationic solutes with sodium dodecyl sulfate on the formation of micelles and critical micelle concentration. AB - The migration behavior of cationic solutes and influences of the interactions of cationic solutes with sodium dodecyl sulfate (SDS) on the formation of micelles and its critical micelle concentration (CMC) were investigated by capillary electrophoresis at neutral pH. Catecholamines and structurally related compounds, including epinephrine, norepinephrine, dopamine, norephedrine, and tyramine, which involve different extents of hydrophobic, ionic and hydrogen-bonding interactions with SDS surfactant, are selected as cationic solutes. The dependence of the effective electrophoretic mobility of cationic solutes on the concentration of surfactant monomers in the premicellar region provides direct evidence of the formation of ion-pairs between cationic solutes and anionic dodecyl sulfate monomers. Three different approaches, based on the variations of either the effective electrophoretic mobility or the retention factor as a function of surfactant concentration in the premicellar and micellar regions, and the linear relationship between the retention factor and the product of a distribution coefficient and the phase ratio, were considered to determine the CMC value of SDS micelles. The suitability of the methods used for the determination of the CMC of SDS with these cationic solutes was discussed. Depending on the structures of cationic solutes and electrophoretic conditions, the CMC value of SDS determined varies in a wide concentration range. The results indicate that, in addition to hydrophobic interaction, both ionic and hydrogen bonding interactions have pronounced effects on the formation of SDS micelles. Ionic interaction between cationic solutes and SDS surfactant stabilizes the SDS micelles, whereas hydrogen-bonding interactions weakens the solubilization of the attractive ionic interaction. The elevation of the CMC of SDS depends heavily on hydrogen-bonding interactions between cationic solutes and SDS surfactant. Thus, the CMC value of SDS is remarkably elevated with catecholamines, such as epinephrine and norepinephrine, as compared with norephedrine. In addition, the effect of methanol content in the sample solution of these cationic solutes on the CMC of SDS was also examined. PMID- 15532560 TI - Use of the new, single-isomer, hexakis(2,3-diacetyl-6-O-sulfo)-alpha-cyclodextrin in acidic methanol background electrolytes for nonaqueous capillary electrophoretic enantiomer separations. AB - The new, single-isomer, sulfated alpha-cyclodextrin, the sodium salt of hexakis(2,3-diacetyl-6-O-sulfo)-alpha-cyclodextrin (HxDAS), was used for the first time in acidic methanol background electrolytes (BGEs) to separate the enantiomers of weak base analytes by nonaqueous capillary electrophoresis (NACE). The concentration dependence of the effective mobilities and separation selectivities followed trends similar to those observed earlier in acidic methanol background electrolytes with heptakis(2,3-diacetyl-6-O-sulfo)-beta cyclodextrin (HDAS) and octakis(2,3-diacetyl-6-O-sulfo)-gamma-cyclodextrin (ODAS). In general, interactions between the weak base analytes and HxDAS were weaker than with HDAS and ODAS. For some of the weak base analytes, separation selectivities observed in acidic aqueous and acidic methanol background electrolytes were complementary to each other, permitting the eventual separation of enantiomers that could not be achieved otherwise. PMID- 15532561 TI - Use of liposomes as a dispersed pseudo-stationary phase in capillary electrophoresis of basic proteins. AB - The suitability of liposome capillary electrophoresis for separating basic proteins at different pH values in the acidic domain was evaluated preparing liposomes consisting of large unilamellar vesicles of the phospholipid 1 palmitoyl-2-oleyl-sn-glycero-3-phosphocholine (POPC), whose hydrodynamic diameter and size distribution were evaluated by dynamic light scattering measurements. The study was conducted evaluating different approaches of performing liposome capillary electrophoresis of proteins, using liposomes dispersed in the electrolyte solution of different composition and pH values ranging from pH 6.2 to 4.0. The liposomes were employed as a pseudo-stationary phase dispersed in the electrolyte solution, which was introduced into the capillary as a plug of volume equivalent to that of the capillary tube, whereas liposome-free electrolyte solutions were contained into the electrolyte vessels during electrophoresis. The study was performed with either bare fused-silica capillaries or capillaries previously treated with POPC that was employed as a dynamic coating agent. Such treatment was performed rinsing the capillary tube with a proper volume of electrolyte solution containing POPC and resulted in a significant reduction of the electroosmotic flow, indicative of the adsorptive behavior of liposomes onto the internal surface of bare fused-silica capillaries. The usefulness of using liposome dynamically coated capillaries for separating basic proteins in absence of the dispersed liposome pseudo-stationary phase filled into the capillary prior to injecting the protein sample was evaluated too. The results show that the presence of liposomes reduced drastically the untoward interactions between basic proteins and the capillary wall, also in capillary not subjected to the dynamic coating process with POPC. In addition, the use of liposomes as a pseudo stationary phase dispersed in the electrolyte solution enhanced the separation of basic proteins also in POPC dynamically coated capillaries. PMID- 15532562 TI - Separation of low-molecular mass peptides by capillary electrophoresis with the use of alkylamines as dynamic coating agents at low pH. AB - Modification of the silanophilic activity of the inner surface of the capillary wall was studied in a capillary electrophoretic system using alkylamines containing background electrolytes at acid pH. The effect of the following amine additives was investigated: (1) alkyl-alpha,omega-diamines (1,2-diaminoethane, 1,4-diaminobutane, 1,7-diaminoheptane, spermine), (2) polymeric amines (polyethylenimine, polybrene), (3) cationic amine surfactants (cetrimide, hexamethonium bromide). A seven membered test mixture of peptides (Gly-Pro-Ala, Pro-hPro, Gly-Pro-Arg, Gly-Pro-Gln, Lys-Pro-Gly, Asn-Pro-Gly, His-Pro-Gly) possessing one or more amino groups was used for selectivity evaluation. Under optimised concentration of the amine modifiers the selectivity was always improved (except for polybrene), particularly with the fast moving analytes. The best results were obtained with 1,2-diaminoethane and 1,7-diaminoheptane. On the other hand with slowly moving peaks the best separations were obtained with 1,7 diaminoheptane, hexadecyltrimethylammonium bromide and hexamethonium bromide, i.e. with modifiers possessing large aliphatic domains which are likely to be hydrophobically bonded with the separated solutes. The selectivity improvement with fast moving members of the test mixture can be ascribed to the decrease of the electroosmotic flow, while the improved separation with slowly moving peaks appears to reflect the altered interaction with the hydrophobized capillary wall. As expected the endoosmotic flow was in all cases decreased. The practical applicability of using amine based dynamic modifiers of the capillary wall was demonstrated on a natural peptide mixture (bacterial collagenase hydrolysate of collagen types I and III). PMID- 15532563 TI - Isolation and characterization of R-phycoerythrin subunits and enzymatic digests. AB - Subunits and enzymatic digests of the highly fluorescent phycobiliprotein R phycoerythrin (R-PE) were analyzed by several separation and detection techniques including HPLC, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE), CE, and HPLC-electrospray ionization (ESI) MS. R-PE subunits were isolated by HPLC and detected as single molecules by total internal reflection fluorescence microscopy. The results show efficient absorption and fluorescence of the R-PE subunits and digest peptides, originating from the incorporation of phycoerythrobilin and phycourobilin chromophores in them. In addition, HPLC-ESI MS and SDS-PAGE were optimized to determine the molecular masses of phycobiliprotein subunits and the chromophore-containing peptides, as well as the amino acid sequences of the latter. Favorable spectroscopic and structural properties of R-PE subunits and enzymatic digests, even under denaturing conditions, make these molecules suitable for use as fluorescence labels for biomolecules. PMID- 15532564 TI - High-throughput protein analysis by multiplexed sodium dodecyl sulfate capillary gel electrophoresis with UV absorption detection. AB - We have developed a novel, high-throughput approach for molecular mass determination of proteins from 14 200 to 116 000 based upon multiplexed, absorbance-based capillary electrophoresis. Via capillary multiplexing, 96 samples were analyzed simultaneously within 30 min. Detection with ultraviolet light obviates the need for protein staining or derivatization. The detection limit of the system was estimated at 5 microg/ml bovine serum albumin (BSA) when sampled from 12.5 mM Tris-HCl. The linear dynamic range was over two orders of magnitude from 5 microg/ml to 1000 microg/ml for BSA. Better than 5% sizing accuracy for protein molecular mass determination and excellent run-to-run and day-to-day reproducibility was obtainable with the described method. PMID- 15532565 TI - Monitoring of the conjugation reaction between human serum transferrin and fluorescein isothiocyanate by capillary electrophoresis. AB - Labeling of iron-free human serum transferrin by an amine-reactive probe, fluorescein isothiocyanate (FITC) was monitored with different dye to protein ratios. The degree of labeling was followed and determined by capillary electrophoresis. Depending on the number of the bound FITC, a shift in the electropherogram was observed, but the conjugated-transferrin forms were resolved from the unbound FITC in zone electrophoresis. The lowest protein concentration that resulted in detectable transferrin conjugate was 0.13 microM, and the limit of detection of the conjugated protein was 0.13 nM (10 ng/ml) with a signal to noise ratio of one to three. PMID- 15532566 TI - Size-based continuous-flow directional control of DNA with a nano-pillar anisotropic array. AB - A size-based pre-fractionation chip applicable for enrichment of macromolecules in biological samples is proposed. In this chip, a fractionation unit with a nano pillar anisotropic array, which functions as sieve meshes, is installed at a bi forked junction. The mesh sizes are close to the diameters of macromolecules and vary according to the direction to the main and branch exit channels. This fractionation unit provides continuous-flow fractionation without both the dilution of samples and the limitations with the sample loading volume. Additionally, the small channel-volume, which is characteristic of chip-based systems, can reduce sample loss. Using nanometer-scale fabrication technology, we fabricated the chips and demonstrated the flow direction control of DNA molecules. PMID- 15532567 TI - Single-step quantitation of DNA in microchip electrophoresis with linear imaging UV detection and fluorescence detection through comigration with a digest. AB - We demonstrate a convenient single-step quantitation technique for double stranded DNA (dsDNA) fragments in polymerase chain reaction (PCR) products based on microchip capillary electrophoresis (micro-CE)/UV or fluorescence detection. PCR products of polymorphisms on the human Y-chromosome related to spermatogenic failure did not need purification. They were premixed and comigrated with a DNA digest whose concentration was known. Hydroxyethyl cellulose (HEC) dissolved in 5x Tris-borate-EDTA (5x TBE, pH 8.3) was used as a separation matrix in a linear polyacrylamide-coated quartz microchip, while mixed poly(ethyl oxides) (PEOs) of different molar-masses dissolved in 1 x TBE (pH 8.3) containing 1 ng/microl ethidium bromide was used as a separation matrix in an uncoated poly(methyl methacrylate) (PMMA) microchip. Elution profiles were monitored under either real time linear imaging UV detection in the snapshot mode where the total separation time is fixed, or light-emitting diode (LED) confocal fluorescence detection in the finishline mode where solutes migrate over the same separation length. It is found that, in both modes, a linear relation exists between the peak areas (A) and the multiplication of the digest concentrations (C) and the fragment sizes (L) in a DNA restrictive digest. Using the comigration electropherogram of a single-step experiment, the concentrations of PCR products were directly determined using the A versus LC linear relationship. The sole condition to obey is that the chosen digest has different fragment sizes with the PCR products of interest. This condition is easy to obey, because micro-CE owns high separation ability, and many digests are commercially available. The recovery of the technique was between 98 and 105%. The R.S.D. for chip-to-chip concentration measurements was less than 6.0% (n = 6). Hence, the technique was accurate and reliable for DNA assays. PMID- 15532568 TI - DNA profiling by capillary array electrophoresis with non-covalent fluorescent labeling. AB - Increasing need for large-scale DNA profiling necessitated the development of automated electrophoresis based methods enabling rapid, high performance analysis of nucleic acids in a wide molecular-mass range. In this paper, we report on the adaptation of a commercial 96-capillary array electrophoresis (CAE) instrument for high-throughput DNA fragment analysis and the evaluation of the effects of different non-covalent DNA staining dyes on separation efficiency. The applicability of different color internal fluorescent standards is shown with mathematical spectral overlap correction algorithms. Large-scale quality control assessment of oligonucleotide probes using non-covalent fluorophore labeling is also demonstrated. The method requires small sample amounts, offers automation and quantification capabilities to accommodate modern biotechnology industry needs. PMID- 15532569 TI - Separation and quantification of viral double-stranded RNA fragments by capillary electrophoresis in hydroxyethylcellulose polymer solutions. AB - Capillary electrophoresis (CE) is an analytical technique widely utilized to resolve complex mixtures of nucleic acids. CE uses a variety of polymers in solution that act as a molecular sieve to separate nucleic acid fragments according to size. It has been shown previously that purified dsDNA can be resolved efficiently by solutions of hydroxyethylcellulose (HEC) polymer, providing a rapid and high resolution method of separation. We have applied this separation technique to viral double-stranded (ds) RNA segments derived from rotavirus process samples. HEC polymers of various molecular masses and concentrations were identified and compared for their ability to separate dsRNA based on the extent of expected polymer network formation. The HEC polymer exhibiting the most desirable separation characteristics was then used for subsequent optimization of various method parameters, such as, injection time, electric field strength, dye concentration and capillary equilibration. The optimized method was then applied to the quantification of genome concentration based on a representative segment of the rotavirus genome. This study demonstrated that purified viral dsRNA material of known concentration could be used to generate an external standard curve relating concentration to peak area. This standard curve was used to determine the concentration of unknown samples by interpolation. This novel RNA quantification assay is likely to be applicable to other types of virus, including those containing dsDNA. PMID- 15532570 TI - Single-stranded DNA-binding protein facilitates gel-free analysis of polymerase chain reaction products in capillary electrophoresis. AB - It has been recently demonstrated that single-stranded DNA-binding protein (SSB) can facilitate quantitative analyses of DNA, RNA, and proteins in gel-free capillary electrophoresis (CE). Here, we report the application of SSB-mediated gel-free CE for analyses of polymerase chain reaction (PCR) products. The unique ability of SSB to bind ssDNA but not double-stranded DNA (dsDNA) allows efficient separation of three types of DNA molecules in the PCR reaction mixture: primers, products (amplified templates), and by-products, which originate from non specific DNA hybridization. SSB-mediated gel-free CE analysis of PCR products combines simplicity, high sensitivity, and outstanding quantitative capabilities. The ability of the method to distinguish between products and by-products makes this method an indispensable tool in preparative PCR (e.g., in the development of nucleotide aptamers). PMID- 15532571 TI - High sensitivity analysis of nitrite and nitrate in biological samples by capillary zone electrophoresis with transient isotachophoretic sample stacking. AB - Tissue level of nitrate and nitrite are established indicators of altered nitric oxide metabolism under various pathological conditions. Determination of these anions in biological samples, in the presence of high chloride concentration, using capillary zone electrophoresis suffers from poor detection sensitivity. Separation conditions providing excellent resolution and submicromolar detection sensitivity of nitrate and nitrite have been developed and validated. Simple sample preparation was applied that maintains nitrite stability in tissue extracts and at the same time allows transient isotachophoresis stacking of the analytes. Nitrate and nitrite concentrations in rat brain and liver tissue samples were determined in control and lipopolysaccharide treated animals. PMID- 15532572 TI - Simultaneous determination of nitrate and nitrite in biological fluids by capillary electrophoresis and preliminary study on their determination by microchip capillary electrophoresis. AB - In order to develop a highly sensitive and high-throughput screening method for nitrogen monoxide metabolites in biological fluids, we have investigated the simultaneous determination of nitrite and nitrate, using capillary electrophoresis and microchip capillary electrophoresis. In capillary zone electrophoresis, a running buffer based on human serum components with high ionic strength has been developed for the determination of nitrite and nitrate in human serum and human saliva. We obtained successful separation of nitrite and nitrate in the serum and the saliva within 7 min under optimum analytical conditions. Linear calibration curves for nitrite and nitrate for both peak height and area were obtained by a standard addition method. The limits of detection obtained at a signal-to-noise ratio (S/N) of 3 for nitrite and nitrate in the serum were 2.6 and 1.5 microM, respectively. The values of the relative standard deviation of peak height for the serum with 9.2 microM nitrite and 20.9 microM nitrate were 5.7 and 4.1%, respectively. For on-site analysis with high-throughput screening, a microchip capillary electrophoresis method using a microchip made of quartz with a UV detector was developed. In this high-throughput format, using a running buffer with an electroosmotic flow modifier, the peaks of nitrite and nitrate in an artificial serum sample were obtained within 8 s. In high-resolution mode, using the buffer without electroosmotic flow modifier, the separation of nitrite and nitrate was obtained within 15 s. In high-resolution mode, using an artificial serum sample with 50 microM NO2- and 50 microM NO3-, the limits of detection (S/N = 3) of 41 microM for NO2- and 26 microM for NO3- were obtained. The method was applied to human serum and saliva. We obtained peaks due to nitrite and nitrate in 10-fold diluted saliva. PMID- 15532573 TI - Simultaneous determination of nitrite, nitrate, thiocyanate and uric acid in human saliva by capillary zone electrophoresis and its application to the study of daily variations. AB - Simultaneous determination of nitrite (NO2-), nitrate (NO3-), thiocyanate (SCN-) and uric acid in human saliva was performed by capillary zone electrophoresis using a coated capillary with reversed electroosmotic flow (EOF), using a 100 mM sodium phosphate buffer at pH 6.5 as a running buffer. Saliva samples were deproteinized with acetonitrile and filtered through a membrane filter. The important advantages of the reported method are: simple operation, short analysis time, minimal sample pre-treatment and sample dilution. In order to evaluate the daily variations of the anionic components, the concentrations were determined in the human saliva of four healthy volunteers upon waking and at 2qh intervals during a day. PMID- 15532574 TI - Evaluation of diabetes-related short-chain organic acids in rat plasma by capillary electrophoresis. AB - A capillary zone electrophoresis method was optimised to analyse low-molecular mass organic acids for the purpose of monitoring diabetes in rat plasma. The method included acetoacetic, 2-hydroxybutyric, lactic and uric acids. A variation in the background electrolyte allowed us to measure pyruvic acid in the same sample. Conditions have been optimised for measuring a large number of plasma samples corresponding to control and diabetic rats. Samples were mixed with acetonitrile (1:1, v/v) to precipitate proteins, centrifuged, diluted and injected. Tropic acid was chosen as an adequate internal standard. Separation was developed with reversed voltage by using a column cartridge pre-treated with polyacrylamide. Two electrophoretic buffers were employed: 0.150 M H3PO4 made up pH 6.20 with NaOH and 0.3 mM CaCl2 for acetoacetic, hydroxybutyric, lactic and uric acids, and 200 mM phosphate-10 mM acetate pH 4.0 for pyruvic acid, both with direct detection at 200 nm. The method was validated for linearity, accuracy and precision and the limits of quantification were calculated. The method was successfully applied to analyse these organic acids in control and diabetic animals. Acetoacetic and hydroxybutyric acids were clearly increased in diabetic rats, meanwhile no statistically significant difference has been found with the other acids. PMID- 15532575 TI - Rapid and direct analysis of gamma-hydroxybutyric acid in urine by capillary electrophoresis-electrospray ionization ion-trap mass spectrometry. AB - The present work was aimed at the development of a capillary electrophoretic analysis of gamma-hydroxybutyric acid (GHB) using electrospray ion trap mass spectrometry to achieve the direct and unequivocal detection of this analyte in human urine. Optimized capillary electrophoretic conditions were: injection, 20 s at 0.5 psi (1 psi = 6894.76 Pa); buffer electrolyte, 12.5 mM ammonium formate adjusted to pH 8.35 with diethylamine; fused silicacapillary: 100 cm x 50 microm i.d.; separation voltage, 25 kV (forward polarity) + 0.5 psi; room temperature. Electrospray and mass spectrometric conditions were: drying gas and nebulizing gas (nitrogen) at flow rate 3 l/min, temperature 250 degrees C, nebulizer pressure: 10 psi; sheath liquid solution: methanol-water (90:10) containing 0.1% ammonia delivered at 3 microl/min; spray voltage 3.5 kV. Mass spetrometric detection was carried out in the selected ion monitoring mode of negative molecular ions at 103 m/z for GHB and 115 m/z for maleic acid (I.S.). Under these conditions the baseline separation of GHB and the I.S. was obtained. The selectivity of the analysis allowed for direct injection of unextracted urine, previously diluted 1:4 with water. Linearity was assessed in the GHB concentration range from 80 to 1280 microg/ml in urine. Analytical sensitivity (as limit of detection) resulted about 5 microg/ml in water and 20 microg/ml in original urine. Analytical precision was fairly acceptable with R.S.D. values lower than 5% for migration times and 18% for quantitation in real samples, in both intra day and day-to-day experiments. On these grounds, the developed method can be adopted for rapid identification of acute intoxications from GHB in humans. PMID- 15532576 TI - Comparison of ion-pair chromatography and capillary zone electrophoresis for the assay of organic acids as markers of abnormal metabolism. AB - The abnormal organic acids in urine are closely related with physiological metabolism. To determinate the low-molecular-mass metabolites in human biological fluids, although there were some previous reports by both of capillary electrophoresis and ion-exchange high-performance liquid chromatography, but it was rarely found by reverse phase of liquid chromatography using ion pair reagent. The objective of this study was aimed to suggest and compare two methods, an additional chromatographic method-ion-pair chromatography (IPC) and a sharp capillary zone electrophoresis (CZE), to determinate organic acids, acting as the abnormal metabolic markers, namely uric acid, orotic acid, pyruvic acid, alpha-ketoglutaric acid, fumaric acid, and hippuric acid. The proposed method of IPC possessed both the extreme stability for column and the good results of reproducibility, linearity and detection limit. The optimum mobile phase was 22% methanol and 10 mM tetra-n-butyl ammonium hydrogen sulfate (pH 4) by gradient elution. As well as the optimum condition of CZE was 5% acetonitrile and 0.5 mM CTAB in phosphate buffer. From the results, CZE showed better recovery and sharp lucid electropherogram. Finally, the two proposed analytical methods were applied to assay human urine with direct and spiked analysis. CZE showed good potency to overcome the sample-to sample variation with standard deviation less than 10%. By comparison results of urinary spiked analysis between IPC and CZE by statistical paired t-test, the results were evaluated no significant difference under P < 0.05. The quantitative linearity of both methods was fitted in application of clinical biological analysis even with 50-fold dilution. PMID- 15532577 TI - Analysis of amino acids by miniaturised isotachophoresis. AB - A method allowing the miniaturised isotachophoretic analysis of amino acids has been developed. To overcome the problems of carbonate contamination which occur when performing separations at alkaline pH levels glycolate was used as the leading ion. Addition of magnesium to the leading electrolyte as a counter species was found to improve the separations. The method has been used on a poly(methyl methacrylate) microdevice with integrated on-column conductivity detectors. The behaviour of a range of common amino acids was investigated and successful separations of up to seven amino acids were made. Good linearity was observed with calibration curves for aspartic acid and phenylalanine over the range 0.063-1.0 mM. Limits of detection for these two species were calculated to be 0.060 and 0.018 mM, respectively. PMID- 15532578 TI - Preconditions for reasonable detection sensitivity and for zone symmetry in electrophoretic separations of cluster borane anions. AB - Single-cage boron cluster anions with at least 11 cluster atoms, free of bonded functional groups that strongly absorb UV light, and their cobalt complexes have been the investigated compounds. Their UV-absorption spectra have absolute maxima between 200 and 215 nm. Corresponding molar extinction coefficients that are of the order of 10(3)-10(4) L mol(-1) cm(-1) indicate medium detection sensitivity. Its reaching requires elimination of background electrolytes that weaken the UV light beam in any way. Frequently used carboxylic acids and zwitterionic Good's buffers cannot be used as buffering compounds from this reason. Freshly prepared 1 mM solutions of boron cluster compounds in aqueous sodium chloride, chosen as indifferent electrolyte, which contain 20-30% (v/v) of methanol or acetonitrile, give zones free of tailing. After storing in the fridge, zones of the compounds became pronouncedly tailed even if their solutions remain clear and free of precipitation, turbidity or opalescence. The tailing usually disappeared if the acetonitrile or the methanol concentration in samples was 40-60% (v/v) depending on the dissolved compound hydrophobicity. Solutions of extremely hydrophobic compounds, stored in the fridge, require mild heating to 30-40 degrees C for half an hour for the avoiding of the tailing. Permanent slow decrease in effective mobilites of boron cluster anions was found if background electrolytes contained acetonitrile and beta-cyclodextrin. Analogous decrease was not observed with organic anions. Constant mobilities of boron cluster anions have been reached if acetonitrile was replaced with methanol. Analyte zones were more symmetrical in background electrolytes buffered with sodium borate of pH 9 than in background electrolytes buffered with sodium phosphate of pH 7. PMID- 15532579 TI - Improved separation of metallothionein isoforms by the presence of cyclodextrin in capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE) with cyclodextrin (CD) in the polyacrylamide coated capillary was used to study metallothionein (MT) forms in the horse kidney preparation produced commercially by Sigma. It is known that CDs form complexes with hydrophobic amino acids. The results show that the presence of CD improves the separability of the various MT forms, including the MT-IA and the MT-IB forms, metallothionein aggregates, as well as the so far unidentified a and b forms. This was true both below and above the isoelectric points (pIs), although the migration times were somewhat longer at increasing CD concentrations for runs at constant voltage than with constant current. PMID- 15532580 TI - Use of a hepta-Tyr antibiotic modified silica stationary phase for the enantiomeric resolution of D,L-loxiglumide by electrochromatography and nano liquid chromatography. AB - Hepta-Tyr antibiotic modified silica stationary phase was used for the chiral resolution of D,L-loxiglumide, a new drug under investigation proposed for the treatment of gastrointestinal diseases. The chiral stationary phase was packed into fused silica capillaries of 75 microm i.d. for a length of only 7 cm and used for both capillary electrochromatography (CEC) and nano-liquid chromatography (nano-LC) running the experiments with the same instrumentation; in order to increase the electroosmotic flow (EOF) the antibiotic stationary phase was mixed with amino-silica particles (3:1, w/w) generating a relatively high reversed EOF. The enantiomeric resolution of loxiglumide by CEC was strongly influenced by several experimental parameters such as applied electric field, mobile phase composition, capillary temperature, etc. Optimum experimental conditions were found applying 15 kV at 20 degrees C and eluting with acetonitrile-sodium phosphate buffer at pH 6 (1:1, v/v). The same capillary was tested for nano-LC experiments. Good chiral separation of loxiglumide was achieved selecting the appropriate mobile phase considering the type and concentration of organic modifier. The nano-LC optimised method was therefore validated and applied to the analysis of a pharmaceutical formulation declared to contain only D-loxiglumide. PMID- 15532581 TI - Enantioselective determination of the novel antidepressant mirtazapine and its active demethylated metabolite in human plasma by means of capillary electrophoresis. AB - Mirtazapine is a recent noradrenergic and specific serotonergic antidepressant drug. A capillary electrophoretic method has been developed for the enantioseparation and analysis of mirtazapine and its main active metabolite, N desmethylmirtazapine, in human plasma. For method optimisation several experimental parameters were investigated, such as type and concentration of the chiral selector, buffer pH and capillary temperature. Baseline enantioseparation of the analytes was achieved in 2.5 min in a fused silica capillary (50 microm i.d.; 48.5 cm total length; 8.5 cm effective length) using carboxymethyl-beta cyclodextrin, dissolved in a background electrolyte consisting of 50 mM phosphate buffer at pH 2.5, as the chiral selector. UV detection was set at 205 nm. A careful pre-treatment of plasma samples was developed, using solid-phase extraction with hydrophilic-lipophilic balance cartridges (60 mg, 3 mL), eluting the sample with methanol, then concentrating it 37.5 times before injection. Extraction yield values are very satisfactory, being the average 89% for mirtazapine and 73% for N-desmethylmirtazapine. Application of the method to some human plasma samples has given satisfactory results. PMID- 15532582 TI - Screening of inhibitors of uridine diphosphate glucuronosyltransferase with a miniaturized on-line drug-metabolism system. AB - Inhibition of uridine diphosphate glucuronosyltransferase (UGT), a major drug metabolyzing enzyme, has been studied using an on-line drug-metabolism system integrated into capillary electrophoresis. Microsomes isolated from rat liver were encapsulated in tetramethoxysilane (TMOS)-based silica matrices within a capillary in a single step under mild conditions. This microsome-immobilized capillary column allows both the metabolism of drugs and determination of the metabolites in a single capillary simultaneously, just by injecting the substrate coenzyme mixture onto the column. Glucuronidation of acetaminophen, a widely used pharmaceutical analgesic and antipyretic agent, was investigated using this system. The glucuronidation was inhibited by 4-nitrophenol (4NP) or probenecid that was injected onto a column along with the substrate-coenzyme mixture. On the other hand, valproate did not inhibit the metabolizing reaction. The extents of inhibition using encapsulated UGT were almost the same as those obtained using free UGT. On the other hand, this electrophoretic enzyme-inhibitor assay in microfabricated devices consumes 10(4) less sample and 10(3) less microsome per experiment compared to the conventional reaction schemes. These results demonstrate that this on-line system can circumvent laborious procedures for the isolation and determination of drug metabolites from the reaction mixtures required in the conventional schemes and can provide an attractive alternative technique for the analysis of drug interactions in the metabolic pathways. PMID- 15532583 TI - Determination of ciprofloxacin and its impurities by capillary zone electrophoresis. AB - Capillary zone electrophoresis (CZE) has been elaborated for separation, identification and determination of ciprofloxacin and its impurities. The separation, phosphate buffer pH 6.0 was supplemented with 0.075 M pentane-1 sulfonic acid sodium salt. The elaborated method was validated. The selectivity, linearity, limits of detection (LOD) and quantification (LOQ), precision, and accuracy of capillary zone electrophoresis were evaluated. The results obtained by CZE were also compared with those obtained by liquid chromatography. Regarding the validation results the CE method fulfils the current European Pharmacopoeia (Eur. Ph.) requirements. The evaluated CE method could be applicable to the analysis of different medicinal products containing ciprofloxacin. PMID- 15532584 TI - Identification of new oxycodone metabolites in human urine by capillary electrophoresis-multiple-stage ion-trap mass spectrometry. AB - Capillary electrophoresis-electrospray ionization multiple-stage ion-trap mass spectrometry (CE-MSn) and computer simulation of fragmentation are demonstrated to be effective tools to detect and identify phase I and phase II metabolites of oxycodone (OCOD) in human urine. OCOD is a strong analgesic used for the management of moderate to severe mainly postoperative or cancer-related pain whose metabolism in man is largely unknown. Using an aqueous pH 9 ammonium acetate buffer and CE-MSn (n < or = 5), OCOD and its phase I metabolites produced by O-demethylation, N-demethylation, 6-ketoreduction and N-oxidation (such as oxymorphone, noroxycodone, noroxymorphone, 6-oxycodol, nor-6-oxycodol, oxycodone N-oxide and 6-oxycodol-N-oxide) and phase II conjugates with glucuronic acid of several of these compounds could be detected in alkaline solid-phase extracts of a patient urine that was collected during a pharmacotherapy episode with daily ingestion of 240-320 mg of OCOD chloride. The data for three known OCOD metabolites for which the standards had to be synthesized in-house, 6-oxycodol, nor-6-oxycodol and oxycodone-N-oxide, were employed to identify two new metabolites, the N-oxidized derivative of 6-oxycodol and an O-glucuronide of this compound. CE-MSn and computer simulation of fragmentation also led to the identification of the N-glucuronide of noroxymorphone, another novel OCOD metabolite for which no standard compound or mass spectra library data were available. PMID- 15532586 TI - Determination of folic acid in tablets by microemulsion electrokinetic chromatography. AB - A microemulsion electrokinetic chromatography (MEEKC) method has been developed and validated for the determination of folic acid, a water-soluble vitamin, in a commercial tablet formulation. The analysis was performed using a microemulsion containing 0.5% (w/w) ethyl acetate, 1.2% (w/w) butan-1-ol, 0.6% (w/w) sodium dodecyl sulfate, 15% (v/v) 2-propanol and 82.7% (w/w) 10 mmol L(-1) sodium tetraborate aqueous buffer at pH 9.2. Direct UV detection at 214nm led to an adequate sensitivity without interference from sample excipients. For quantitative purposes, niacin was used as internal standard. Acceptable precision (<1.2% relative standard deviation (R.S.D.)), linearity (r = 0.9992; range from 160.0 to 240.0 microg/mL), sensitivity (limit of detection (LOD) = 2.98 microg/mL; limit of quantification (LOQ) = 9.05 microg/mL) and recovery (99.8 +/- 1.8% at three concentration levels) were obtained. Based on the performance characteristics, the proposed methodology was found suitable for the determination of folic acid in tablet formulations. PMID- 15532585 TI - Electrophoretic behavior and pKa determination of quinolones with a piperazinyl substituent by capillary zone electrophoresis. AB - Electrophoretic behavior and pKa determination of six quinolones with a piperazinyl substituent, together with two quinolones without a piperazinyl substituent and 1-phenylpiperazine, were investigated by capillary zone electrophoresis. The results indicate that quinolones with a piperazinyl substituent involve three protonation/deprotonation equilibria. The results also suggest that the contribution of the zwitterionic species of these quinolones to the effective mobility may not be neglected. This is probably due to a slightly incomplete protonation of the piperazinyl moiety in the pH range of 6.0-8.0, compared with the complete dissociation of the carboxylic group. Consequently, the zwitterionic species of ciprofloxacin, in particular, is slightly negatively charged. With the aid of computer simulation, three pKa values were determined for quinolones with a piperazinyl substituent, thus allowing us to rationalize precisely the influence of pH on the electrophoretic behavior of these compounds. PMID- 15532587 TI - Simultaneous separation and on-line concentration of amitrole and benzimidazole pesticides by capillary electrophoresis with a volatile migration buffer applicable to mass spectrometric detection. AB - This study used capillary electrophoresis (CE) to investigate the simultaneous separation and on-line concentration of five pesticides: amitrole (AMT), carbendazim (MBC), 2-aminobenzimidazole (ABI), thiabendazole (TBZ) and 1,2 diaminobenzene (DAB). A volatile migration buffer was used for the investigation because of the applicability to mass spectrometric (MS) detection. They were separated completely at pH 4.0 as a result of changing pH using formic acid ammonium formate buffer. Values of the dissociation constant for MBC and DAB estimated from the changes in the mobility with pH showed good agreement with values in the literature. Dissociation constants for AMT and TBZ were estimated. Limits of detection (LODs) for the analytes were on the ppm level with UV detection under the optimized separation condition. On-line concentration by simple stacking mode was not effective except to 2-aminobenzimidazole because of the peak tailing. The addition of formic acid to sample matrix improved the peak shapes. That improvement may be attributed to transient isotachophoretic effect. The concentration factors obtained from the comparison of the LODs were in the range of 7.6-27-fold. This concentration method was applied preliminarily to CE with MS detection. PMID- 15532588 TI - Determination of volatile corrosion inhibitors by capillary electrophoresis. AB - In this work, a capillary electrophoresis (CE) method using indirect UV detection (214nm) for the simultaneous determination of monoethanolamine (MEA), diethanolamine (DEA), triethanolamine (TEA), diethylethanolamine (DEEA), monocyclohexylamine (MCHA) and dicyclohexylamine (DCHA) in water/ethanol extracts of wrapping materials containing volatile corrosion inhibitors (VCIs) was described. A running buffer consisting of 0.010 molL(-1) imidazole, 0.010 molL( 1) 2-hydroxyisobutyric acid (HIBA) and 0.010 molL(-1) 18-crown-6 ether enabled separation of the analytes in less than 7 min. A few method validation parameters were determined revealing good migration time repeatability (<0.7% RSD) and area repeatability (< 1.8% RSD). Limits of detection were in the range of 0.52-1.54 mg L(-1). Recovery values were in the range of 94.8-100.9%. The methodology was successfully applied to the analysis of three commercial products (VCI treated paper, foam and plastic). The concentration of amines in these materials varied from 0.050 to 22.3% (w/w). PMID- 15532589 TI - Men's surveys: new findings. PMID- 15532590 TI - [Clinical and immunological characteristics of HIV-positive AIDS in children in Northern Israel]. AB - We are treating 11 children in our AIDS clinic. All were infected by vertical transmission from carrier mothers. However, among 31 HIV-carrier AIDS patients who were under follow-up during pregnancy, supposedly taking zidovudine prophylaxis, only 1 (3.3%) gave birth to a baby infected with HIV. Our children with HIV and AIDS are 3 months to 12 years of age (average 4.5 years); mean age at diagnosis was 18 months. All are either symptomatic or have laboratory evidence of progressive immunodeficiency, 1 is asymptomatic (N2), 1 has mild symptoms (A2) and the rest present significant symptoms or AIDS-defining disease. All have moderate to severe immunodeficiency, as evidenced by CD4+ cells counts. 60% have rapidly progressive disease, based on their symptomatology and immune state, whereas clinical reports in the literature point to only 10-15%. However, the average CD4 + cell count was 22% (749/mm3) at diagnosis and 22% (759/mm3) at last follow-up. These stable findings during an average follow-up of 28 months probably reflect the effect of medical and supportive treatment. All received antiretroviral medication consisting of a combination of 2 or 3 drugs. 8 of 11 also received prophylactic treatment against opportunistic infections and 8 of 11 are clinically well. Routine follow-up and a good relationship with the patient's family increase cooperation and promote optimal medical treatment, and consequently improve the clinical condition and quality of life. PMID- 15532591 TI - [Smoking and incidence of lung cancer, 1981-1995]. AB - Smoking is the dominant risk factor for lung cancer. We compared trends in smoking with those of the incidence of lung cancer in Israel. The proportion of smokers has declined during the past 20 years; the decrease is greater in men than in women, and more marked in the elderly. Since 1980 the age-adjusted incidence of lung cancer in Jewish men has decreased slightly, but in women it has remained constant. Among Arab men there was an increase in age-adjusted incidence of lung cancer and since 1986 it has been higher than in Jewish men. The largest decrease in lung cancer incidence was among Jewish men aged 75 and over. This may be explained by data on the age of smoking cessation in the population. It was observed that the main decrease in smoking occurred among men over the age of 55 in the past 20 years, which correlates with the decline in lung cancer observed in the older age group. Lung cancer rates in Israel are lower than in other western countries despite the similar prevalence of smoking, for unknown reasons. PMID- 15532592 TI - [Irradiation injury of the salivary glands]. AB - Ionizing irradiation of the salivary glands often leads to severe histological and functional alterations. Such exposure usually occurs during radiotherapy in patients with head and neck malignancy. The consequent xerostomia, often life long, may result from even relatively low dosage irradiation, and causes a great deal of suffering. We suggest a radiobiological mechanism for this phenomenon which has been studied extensively since first described in 1911. The suggested injurious role of redox active transition metal ions and highly destructive free radicals is discussed in relation to the ultimate radiosensitive cellular target, DNA. PMID- 15532593 TI - [Plagiocephaly in children: etiology, differential diagnosis and helmet treatment]. AB - Plagiocephaly in a head-and-neck irradiated rat model or rhomboid-shaped head, occurs in at least 1 in 300 live births. In most cases such asymmetry is not caused by synostosis of the unilateral coronal or lambdoid sutures, but is rather a deformity produced by intrauterine and/or postnatal deformational forces. Categorization and diagnosis of plagiocephaly as synostotic or deformational is reliably made by physical examination and computerized tomography. Its differential diagnosis is extremely important because prompt surgical correction is usually indicated for the synostotic type. In contrast, infants with deformational frontal or occipital plagiocephaly generally respond to helmet treatment. 10 infants with significant deformational plagiocephaly were treated with individual plastic helmets during the past 2 years and 4 other infants with plagiocephaly are currently being treated. In each instance, cranial asymmetry dramatically improved as the brain grew and the head filled out the helmet. There were no significant complications. Awareness of deformational plagiocephaly allows more accurate diagnosis and appropriate treatment, avoiding unnecessary surgical intervention in patients with positional molding. PMID- 15532594 TI - [Oral manifestations of bulimia nervosa]. AB - Bulimia nervosa (BN) is an eating disorder frquently accompanied by changes of the hard and soft tissues of the oral cavity and of the salivary glands. Routine dental examination in a 25-year-old woman disclosed oral signs and symptoms characteristic of BN and she was referred for psychiatric evaluation. Oral evaluation of patients suspected of BN is recommended to prevent irreversible damage to the oral and perioral structures. PMID- 15532595 TI - [CT-guided excision of osteoid osteoma]. AB - CT-guided excision of osteoid osteoma is a new surgical technique that enables accurate resection of the nidus during 1-day hospitalization. We present 5-year results in 42 patients (26 males and 16 females, mean age 18 years, range 3-46). In 40 out of 42, complaints disappeared immediately after the procedure. The recovery period was short and the return to normal activity was faster than in the open surgical approach. Complications were minimal and transient. PMID- 15532596 TI - [Malignant lymphoma of the bladder]. AB - Urinary tract lymphoma is usually reported when the secondarily stem is affected by widespread non-Hodgkin lymphoma. We describe an 83-year-old woman who presented with secondary lymphoma of the bladder 3 years after diagnosis when it initially infiltrated her breast. Treatment included local transurethral excision followed by chemotherapy, during which she died of disseminated disease. PMID- 15532597 TI - [Acute malaria in an Israeli tourist to Kenya]. AB - Malaria is 1 of the main causes of death in third world countries. It has become extinct in Israel and imported cases are rare, since most visitors to endemic countries take anti-malarial prophylaxis. We report an Israeli tourist to Kenya infected with falciparum malaria complicated by severe metabolic acidosis, renal failure and adult respiratory distress syndrome. After intensive care treatment this preventable condition improved. PMID- 15532598 TI - [Lung-cancer and smokers in Israel, 1981-1995]. PMID- 15532599 TI - [Evidence-based medicine--perspectives and problems]. PMID- 15532600 TI - [Evidence based medicine--a new era in medicine?]. PMID- 15532601 TI - [Intrauterine metabolism and fetal growth--long term consequences]. PMID- 15532602 TI - [Ocular manifestations of pregnancy and effect on eye disease]. PMID- 15532603 TI - [NMDA receptor antagonists: a new treatment for neuropathic pain]. PMID- 15532604 TI - [Physical examination of sportsmen--the Israeli sports law]. PMID- 15532605 TI - [Chronic oral infections and the development of cardiovascular disease]. PMID- 15532606 TI - [Biopsy of the posterior segment of the eye]. PMID- 15532607 TI - [Penicillin allergy: an update. Part 2: Desensitization to beta-lactam drugs]. PMID- 15532609 TI - [The "bionic tetraplegic": is rehabilitation medicine also becoming too technological?]. PMID- 15532608 TI - [Adverse reactions to minocycline treatment of acne vulgaris]. PMID- 15532610 TI - [Therapeutic effectiveness of sepulchers of saints and holy men--a comparative study]. PMID- 15532611 TI - [Relationship between exposure to formaldehyde and symptoms of irritation during human-dissection course at Chiba University]. PMID- 15532612 TI - Departments of Surgery in South Africa--legacies of the past, challenges for the future. PMID- 15532613 TI - The Department of Surgery: Stellenbosch University. PMID- 15532615 TI - Is the incidence of gastroschisis rising in South Africa in accordance with international trends? A retrospective analysis at Pretoria Academic and Kalafong Hospitals, 1981-2001. AB - OBJECTIVES: Analysis of the incidence of gastroschisis compared with the incidence of exomphalos as a percentage of total paediatric surgical admissions. DESIGN: Retrospective observational analysis using data from the ward admissions registers of the paediatric surgical wards of Pretoria Academic and Kalafong hospitals and from the weekly statistics sheets and audits thereof of the Paediatric Surgery Department at Pretoria Academic Hospital. Patient files from Pretoria Academic were used where available to confirm data. SETTING: Pretoria Academic Hospital (PAH) and Kalafong Hospital (KH), Pretoria, from March 1981 to December 2001, excluding Kalafong Hospital data from February 1984 to 12 June 1984 and for 2001. RESULTS: Forty-eight cases of gastroschisis and 139 cases of exomphalos were seen at PAH and KH out of 21 495 total paediatric surgery ward admissions. The average incidence of gastroschisis increased 35-fold from the 7 year period 1981 - 1987 to the 7-year period 1995 - 2001, while the average incidence of exomphalos compared across the same periods only showed a 1.82-fold increase. The incidence of gastroschisis rose above that of exomphalos in 1999 and remained so through to 2001. The incidence of exomphalos showed a general rising trend from 1981 to 2001 except for a sudden unexplained increase in 1995 and 1996. Gastroschisis presenting at PAH was associated with a mortality rate of at least 38.7%. Reasons for this high mortality rate cannot be adequately evaluated owing to poor record keeping. CONCLUSIONS: The incidence of gastroschisis presenting at PAH and KH has risen dramatically relative to the incidence of the macroscopically similar exomphalos over the period 1981 - 2001, with the incidence of gastroschisis rising above the incidence of exomphalos. A larger scale study looking at birth prevalence of gastroschisis and exomphalos in South Africa is necessary to determine whether this represents a true differential change in the incidence of gastroschisis, reflecting internationally observed increases in the birth prevalence of this defect, or whether it is merely due to logistical changes within the administration of the Department of Health serving the area. PMID- 15532614 TI - Femoro-distal bypass surgery at Groote Schuur hospital--4-year retrospective study. AB - An analysis of the demographics and outcome of femoro-distal bypass in patients presenting with defined critical limb ischaemia at Groote Schuur Hospital, Cape Town, is presented. MATERIALS AND METHODS: A retrospective review was conducted between January 1998 and December 2001. During this period, 65 patients underwent femoro-distal bypass. Of these, 57 patients were analysed and 8 patients were excluded from the study because of incomplete medical records RESULTS: The median age of the patients in this study was 62 years, with a male-to-female ratio of 34:23. Twenty-eight patients (49%) were diabetic. An overall 2-year mortality of 19.2% was recorded. Reversed saphenous vein graft (RSVG) was used in 29 legs (50.9%) and in situ vein graft (ISVG) in 19 legs (33.3%). There was no statistically significant difference in the graft patency between the two methods (p = 0.39); the 2-year cumulative patency was 40% for the RSVG and 39% for the ISVG. In our unit only 2 factors influenced the outcome of femoro-distal bypass- local sepsis in the foot and an increased early postoperative ankle-brachial index (p < 0.05). Diabetes mellitus, gender, age and race had no influence on the outcome. This study showed that the site of the distal anastomosis had no effect on graft patency or limb salvage. At 2 years, the cumulative patency and limb salvage rates were 40% and 56% respectively. Of the 11 grafts with stenotic lesions requiring intervention, 6 were detected between 18 and 24 months. CONCLUSION: Management of critical limb ischaemia is a major part of the workload in our unit, with most patients undergoing primary amputation. The surgical outcome of femoro-distal bypass was largely influenced by local sepsis and early postoperative ABI. We found that more than half the stenotic lesions detected during graft surveillance occurred beyond 18 months postoperatively. This suggests that a graft surveillance programme should continue beyond 18 months. PMID- 15532616 TI - Results of treatment in children with anorectal malformations in Calabar, Nigeria. AB - Leading symptoms of anorectal malformation in the neonatal period are abdominal distention, non-passage of meconium and constipation. When present, vomiting is a late symptom. In a study in Calabar, Nigeria, patients were observed to present late, and teenage mothers in rural communities were mostly affected. Female neonates were affected more than males in a ratio of 1.5:1. Classification into low and high abnormality was adopted and proved practical in terms of identification of the pathology and treatment of the lesion. Patients with low abnormality (N = 24, 44.4%) were treated with perineal cut-down, while those with high abnormality (55.6%) had initial palliative colostomy before a definitive abdominal perineal pull-through procedure. Faecal incontinence (13%), anal stenosis (11.1%), constipation (7.4%) and colostomy prolapse (5.6%) were noted to be associated complications. Poverty and ignorance were noted to be the main factors affecting treatment outcome. A concerted public enlightenment campaign is therefore required. PMID- 15532617 TI - A novel method of removing a foreign body from the external ear canal. PMID- 15532620 TI - The effects of single and double hearing protection on the localization and segregation of spatially-separated speech signals. PMID- 15532622 TI - The influence of sexual orientation on vowel production. PMID- 15532623 TI - Source parameter estimates of echolocation clicks from wild pygmy killer whales (Feresa attenuata). PMID- 15532650 TI - A phenomenological model of peripheral and central neural responses to amplitude modulated tones. AB - A phenomenological model with time-varying excitation and inhibition was developed to study possible neural mechanisms underlying changes in the representation of temporal envelopes along the auditory pathway. A modified version of an existing auditory-nerve model [Zhang et al., J. Acoust. Soc. Am. 109, 648-670 (2001)] was used to provide inputs to higher hypothetical processing centers. Model responses were compared directly to published physiological data at three levels: the auditory nerve, ventral cochlear nucleus, and inferior colliculus. Trends and absolute values of both average firing rate and synchrony to the modulation period were accurately predicted at each level for a wide range of stimulus modulation depths and modulation frequencies. The diversity of central physiological responses was accounted for with realistic variations of model parameters. Specifically, enhanced synchrony in the cochlear nucleus and rate-tuning to modulation frequency in the inferior colliculus were predicted by choosing appropriate relative strengths and time courses of excitatory and inhibitory inputs to postsynaptic model cells. The proposed model is fundamentally different than others that have been used to explain the representation of envelopes in the mammalian midbrain, and it provides a computational tool for testing hypothesized relationships between physiology and psychophysics. PMID- 15532651 TI - Simultaneous measurement of middle-ear input impedance and forward/reverse transmission in cat. AB - Reported here is a technique for measuring forward and reverse middle-ear transmission that exploits distortion-product otoacoustic emissions (DPOAEs) to drive the middle ear "in reverse" without opening the inner ear. The technique allows measurement of DPOAEs, middle-ear input impedance, and forward and reverse middle-ear transfer functions in the same animal. Intermodulation distortion in the cochlea generates a DPOAE at frequency 2f1-f2 measurable in both ear-canal pressure and the velocity of the stapes. The forward transfer function is computed from stapes velocities and corresponding ear-canal pressures measured at the two primary frequencies; the reverse transfer function is computed from velocity and pressure measurements at the DPOAE frequency. Middle-ear input impedance is computed from ear-canal pressure measurements and the measured Thevenin equivalent of the sound-delivery system. The technique was applied to measure middle-ear characteristics in anesthetized cats with widely opened middle ear cavities (0.2-10 kHz). Stapes velocity was measured at the incudo-stapedial joint. Results on five animals are reported and compared with a published middle ear model. The measured forward transfer functions and input impedances generally agree with previous measurements, and all measurements agree qualitatively with model predictions. The reverse transfer function is shown to depend on the acoustic load in the ear canal, and the measurements are used to compute the round-trip middle-ear gain and delay. Finally, the measurements are used to estimate the parameters of a two-port transfer-matrix description of the cat middle ear. PMID- 15532652 TI - Distortion product otoacoustic emission (DPOAE) input/output functions and the influence of the second DPOAE source. AB - Distortion product otoacoustic emissions (DPOAEs) at 2f1-f2 (f2/f1 = 1.2) have two components from different cochlear sources, i.e., a distortion component generated near f2 and a reflection component from the characteristic site of fDP. The interaction of the two sources may negatively affect the DPOAE input/output (I/O) functions that are used to predict either auditory thresholds or the compression characteristics of the basilar membrane. This study investigates the influence of the reflection component on DPOAE I/O functions in a frequency range for f2 from 1500 to 4500 Hz in steps of 18 Hz. A time windowing procedure is used to separate the components from the two DPOAE sources. With decreasing stimulus level, the relative contribution of the reflection component increases. I/O functions from the separated distortion component (DCOAE I/O functions) only show smooth changes in shape and slope with frequency, while "standard" DPOAE I/O functions show rapid changes between adjacent frequencies, indicating a strong influence from the interference with the second DPOAE source. A reduced variability for adjacent frequencies can be seen as well for prediction of hearing thresholds, when using DCOAE instead of DPOAE I/O functions. PMID- 15532653 TI - Searching for the optimal stimulus eliciting auditory brainstem responses in humans. AB - This study examines auditory brainstem responses (ABR) elicited by rising frequency chirps. Two chirp stimuli were developed and designed such as to compensate for cochlear travel-time differences across frequency, in order to maximize neural synchrony. One chirp, referred to as the O-chirp, was based on estimates of human basilar membrane (BM) group delays derived from stimulus frequency otoacoustic emissions (SFOAE) at a sound pressure level of 40 dB [Shera and Guinan, in Recent Developments in Auditory Mechanics (2000)]. The other chirp, referred to as the A-chirp, was derived from latency functions fitted to tone-burst-evoked ABR wave-V data over a wide range of stimulus levels and frequencies [Neely et al., J. Acoust. Soc. Am. 83(2), 652-656 (1988)]. In this case, a set of level-dependent chirps was generated. The chirp-evoked responses, particularly wave-V amplitude and latency, were compared to click responses and to responses obtained with the original chirp as defined in Dau et al. [J. Acoust. Soc. Am. 107(3), 1530-1540 (2000)], referred to here as the M-chirp since it is based on a (linear) cochlea model. The main hypothesis was that, at low and medium stimulation levels, the O- and A-chirps might produce a larger response than the original M-chirp whose parameters were essentially derived from high level BM data. The main results of the present study are as follows: (i) All chirps evoked a larger wave-V amplitude than the click stimulus indicating that for the chirps a broader range of spectral components contributes effectively to the ABR. (ii) Only small differences were found between the O-chirp and M-chirp responses at low and medium levels. This indicates that SFOAE may not provide a robust estimate of BM group delay, particularly at low frequencies, or that frequency-dependent neural delays exist which are not reflected in the design of these chirps. (iii) The A-chirp produced the largest responses, particularly at low stimulation levels. This chirp might therefore be valuable for clinical applications, particularly in tests where the click stimulus has been used so far. PMID- 15532654 TI - Using statistical decision theory to predict speech intelligibility. III. Effect of audibility on speech recognition sensitivity. AB - The speech recognition sensitivity (SRS) model [H. Musch and S. Buus, J. Acoust. Soc. Am. 109, 2896-2909 (2001)] is a macroscopic model for predicting speech intelligibility. The present study proposes a modification to the relation between the SRS model's audibility-noise variance and the signal-excitation to noise-excitation ratio (SNRE) in the auditory periphery. The modified relation is derived from data obtained in nine studies that measured normal-hearing listeners' consonant-recognition performance at several levels of speech-spectrum shaped noise. When the audibility-noise variance is directly proportional to the relative power of the noise excitation in the auditory periphery, the SRS model yields good predictions of the data. Four of the nine studies also reported consonant-recognition performance in various filtering conditions. Good predictions of these data were achieved with SRS model parameters that were consistent with the model parameters fitting the speech-in-noise data and with the model parameters used in the original SRS papers. PMID- 15532655 TI - Informational masking in hearing-impaired and normal-hearing listeners: sensation level and decision weights. AB - Informational masking (IM) refers to elevations in signal threshold caused by masker uncertainty. The purpose of this study was to investigate two factors expected to influence IM in hearing-impaired listeners. Masked thresholds for a 2000-Hz signal in the presence of simultaneous multitone maskers were measured in 16 normal-hearing (NH) and 9 hearing-impaired (HI) listeners. The maskers were 70 dB SPL average total power and were comprised of fixed-frequency components between 522 and 8346 Hz that were separated from each other by at least 1/3 oct and from the signal by at least 2/3 octs. Masker uncertainty was manipulated by randomly presenting each masker component with probability p = 0.1,0.2,...,0.9, or 1.0 across different trial blocks. Energetic masking was estimated as the amount of masking for p = 1.0, where masker uncertainty was minimum. IM was estimated as the amount of masking in excess of energetic masking. Decision weights were estimated by a regression of the listener's yes/no responses against the presence or absence of the signal and masker components. The decision weights and sensation levels (SLs) of the stimulus components were incorporated as factors in a model that predicts individual differences in IM based on the level variance (in dB) at the output of independent auditory filters [Lutfi, J. Acoust. Soc. Am. 94, 748-758 (1993)]. The results showed much individual variability in IM for the NH listeners (over 40 dB), but little IM for most HI listeners. When masker components were presented to a group of NH listeners at SLs similar to the HI listeners, IM was also similar to the HI listeners. IM was also similar for both groups when the level per masker component was 10 dB SL. These results suggest that reduced masker SLs for HI listeners decrease IM by effectively reducing masker variance. Weighting efficiencies, computed by comparing each listener's pattern of weights to that of an ideal analytic listener, were a good predictor of individual differences in IM among the NH listeners. For the HI listeners weighting efficiency and IM were unrelated because of the large variation in masker SLs among individual listeners, the small variance in IM, and perhaps because broadened auditory filters in some listeners increased the covariance in auditory filter outputs. PMID- 15532656 TI - Masker phase effects in normal-hearing and hearing-impaired listeners: evidence for peripheral compression at low signal frequencies. AB - The presence of cochlear-based compression at low frequencies was investigated by measuring phase effects in harmonic maskers. In normal-hearing listeners, the amount of masking produced depends strongly on the phase relationships between the individual masker components. This effect is thought to be determined primarily by properties of the cochlea, including the phase dispersion and compressive input-output function of the basilar membrane. Thresholds for signals of 250 and 1000 Hz were measured in harmonic maskers with fundamental frequencies of 12.5 and 100 Hz as a function of the masker phase curvature. Results from 12 listeners with sensorineural hearing loss showed reduced masker phase effects, when compared with data from normal-hearing listeners, at both 250- and 1000-Hz signal frequencies. The effects of hearing impairment on phase-related masking differences were not well simulated in normal-hearing listeners by an additive white noise, suggesting that the effects of hearing impairment are not simply due to reduced sensation level. Maximum differences in masked threshold were correlated with auditory filter bandwidths at the respective frequencies, suggesting that both measures are affected by a common underlying mechanism, presumably related to cochlear outer hair cell function. The results also suggest that normal peripheral compression remains strong even at 250 Hz. PMID- 15532657 TI - Effects of pulse rate and electrode array design on intensity discrimination in cochlear implant users. AB - The effects of pulse rate on intensity discrimination were evaluated in 14 subjects with Clarion C-I cochlear implants. Subjects had a standard [Clarion spiral electrode array (SPRL group)] or perimodiolar electrode array [Clarion HiFocus electrode array with electrode positioning system (HF+EPS group)]. Weber fractions for intensity discrimination [ Wf(dB)= 10 log deltaI/I] were evaluated at five levels over dynamic range at each of three pulse rates (200, 1625 and 6500 pps) using monopolar stimulation. Weber fractions were smaller for 200 pps stimuli than for 1625 or 6500 pps stimuli in both groups. Weber fractions were significantly smaller for SPRL subjects (mean Wf(dB) = -9.1 dB) than for HF+EPS subjects (mean Wf(dB) = -6.7 dB). Intensity difference limens (DLs) expressed as a percentage of dynamic range (DR) (deltaI%DR= deltaI/DRdB* 100) did not vary systematically with pulse rate in either group. Larger intensity DLs combined with smaller dynamic ranges led to fewer intensity steps over the dynamic range for HF+EPS subjects (average 9 steps) compared to SPRL subjects (average 23 steps). The observed effects of pulse rate and electrode array design may stem primarily from an inverse relationship between absolute current amplitude and the size of intensity DLs. The combination of smaller dynamic ranges and larger Weber fractions in HF+EPS subjects could be the result of increased variability of neural outputs in these subjects. PMID- 15532658 TI - Frequency modulation detection in cochlear implant subjects. AB - Frequency modulation (FM) detection was investigated in acoustic and electric hearing to characterize cochlear-implant subjects' ability to detect dynamic frequency changes and to assess the relative contributions of temporal and spectral cues to frequency processing. Difference limens were measured for frequency upward sweeps, downward sweeps, and sinusoidal FM as a function of standard frequency and modulation rate. In electric hearing, factors including electrode position and stimulation level were also studied. Electric hearing data showed that the difference limen increased monotonically as a function of standard frequency regardless of the modulation type, the modulation rate, the electrode position, and the stimulation level. In contrast, acoustic hearing data showed that the difference limen was nearly a constant as a function of standard frequency. This difference was interpreted to mean that temporal cues are used only at low standard frequencies and at low modulation rates. At higher standard frequencies and modulation rates, the reliance on the place cue is increased, accounting for the better performance in acoustic hearing than for electric hearing with single-electrode stimulation. The present data suggest a speech processing strategy that encodes slow frequency changes using lower stimulation rates than those typically employed by contemporary cochlear-implant speech processors. PMID- 15532659 TI - Masker-first advantage for cues in informational masking. AB - The detectability of a pure-tone signal may be reduced by adding a small number of randomly drawn masker tones remote from the signal frequency, an effect attributed to informational masking. A pretrial cue consisting of either the upcoming signal or masker releases informational masking, but a pretrial cue of the signal-plus-masker stimulus does not. In these experiments the relative potency of pre- and posttrial cues in releasing informational masking was examined. In separate conditions the masker-alone and signal-plus-masker stimuli were cues. The results indicated a masker-first advantage, i.e., sensitivity was superior when a masker cue preceded a yes/no trial interval compared to (a) when a signal-plus-masker preceded the trial, and (b) when either cue type followed the yes/no trial interval. A masker-first advantage was also obtained when the results from a two-interval forced-choice same/different task were examined. In contrast, a masker-first advantage was not obtained when the frequency of the signal to be detected was random. For detection tasks using random multi-tone maskers there may be differences in processing efficiency depending on the order in which stimuli are presented. The "masker-first advantage" may depend, in part, on observers maintaining their attention at the signal frequency. PMID- 15532660 TI - The effect of onset asynchrony on profile analysis by normal-hearing and hearing impaired listeners. AB - The effect of onset asynchrony on discrimination of spectral shape was evaluated for hearing-impaired and normal-hearing listeners. Stimuli were the sum of four tones equally spaced on a logarithmic frequency scale. The standard stimulus had tones of equal amplitude, and the signal stimulus had two adjacent components increased in level, and the other two components decreased in level. Thresholds for discrimination between the standard and signal stimuli were measured as a function of an onset asynchrony among the components of 0, 50, and 200 ms. Hearing-impaired and normal-hearing listeners had similar thresholds when the stimulus components were widely spaced in frequency, but hearing-impaired listeners had much higher thresholds for narrowly spaced components. Excitation pattern analyses indicated that listeners may use spectral peaks in the stimulus rather than the change in excitation across the full stimulus bandwidth for spectral shape discrimination tasks. Increasing temporal asynchrony of components resulted in increased thresholds for both groups of listeners to a greater extent in the wide span than the narrow span. Reduced effects of onset asynchrony in the narrow span suggest that spectral resolvability of components plays an important role in the processing of onset asynchrony across frequency. PMID- 15532661 TI - Enhancing temporal cues to voice pitch in continuous interleaved sampling cochlear implants. AB - The limited spectral resolution of cochlear implant systems means that voice pitch perception depends on weak temporal envelope cues. Enhancement of such cues was investigated in implant users and in acoustic simulations. Subjects labeled the pitch movement of processed synthetic diphthongal glides. In standard processing, noise carriers (simulations) or pulse trains (implant users) were modulated by 400 Hz low-pass envelopes. In modified processing, carriers were modulated by two components: (1) Slow-rate (<32 Hz) envelope modulations, conveying dynamic spectral shape changes crucial for speech; (2) a simplified waveform (e.g., a sawtooth) matching the periodicity of the input diphthong. In both normal listeners and implant users performance was better with modified processing, though temporal envelope cues were less effective with higher F0. Factors contributing to the advantage for modified processing may include increased modulation depth and use of a modulation waveform featuring a rapid onset in each period, resulting in a clearer representation of F0 in the neural firing pattern. Eliminating slow-rate spectral dynamics, so that within-channel amplitude changes solely reflected F0, showed that dynamic spectral variation obscured temporal pitch cues. Though significant, advantages for modified processing were small, suggesting that the potential for developing strategies delivering enhanced pitch perception is limited. PMID- 15532662 TI - Side effects of fast-acting dynamic range compression that affect intelligibility in a competing speech task. AB - Using a cochlear implant simulator, Stone and Moore [J. Acoust. Soc. Am. 114, 1023-1034 (2003)] reported that wideband fast-acting compression led to poorer intelligibility than slow-acting compression in a competing speech task. Compression speed was varied by using different pairs of attack and release times. In the first experiment reported here, it is shown that attack times less than about 2 ms in a wideband compressor are deleterious to intelligibility. In experiment 2, fast wideband compression was applied to the target and background either before or after mixing. The former reduced the modulation depth of each signal but maintained the independence between the two signals, while the latter introduced "comodulation." Using simulations with 6 and 11 channels, intelligibility was higher when compression was applied before mixing. In experiment 3, wideband compression was compared with multichannel compression; the latter led to reduced comodulation effects. For 6 channels, the position of the compressor, either wideband or within each channel, had no effect on intelligibility. For 11 channels, channel compression severely degraded intelligibility compared to wideband compression, presumably because of the greater reduction of across-channel contrasts. Overall, caution appears necessary in the use of fast-acting compression in cochlear implants, so as to preserve intelligibility. PMID- 15532663 TI - Secant lines in a preliminary study of phonetic reduction of /I/ and /epsilon/. AB - Two subjects from the X-Ray Microbeam Speech Production Database were examined in their production of the vowels /I/ and /epsilon/ in alveolar and dental consonant contexts. Secant lines, or first-order splines, between the three most anterior pellets were examined at vowel critical times. These critical times were zero crossings in the tangential acceleration of the midpoints of the secant lines. We expected and found, in general, that vowel reduction occurred as a function of vowel duration in measures of the secant line midpoint-to-palate distance and secant line orientation at vowel critical times. The shorter the vowel, the smaller the distance of the secant line midpoints to the palate and the less downward the orientation of the secant lines at the vowel critical times. Phonetic reduction was also apparent in the formant frequencies. There were differences between the speakers in terms of the range of vowel duration and degree of reduction. The subjects differed in the functional parts of the tongue spanned by the secant lines and the shape of their palates. These differences were factors in the observed relations between formant frequencies and the articulatory, secant line measures for each subject. PMID- 15532664 TI - The distinctness of speakers' productions of vowel contrasts is related to their discrimination of the contrasts. AB - This study addresses the hypothesis that the more accurately a speaker discriminates a vowel contrast, the more distinctly the speaker produces that contrast. Measures of speech production and perception were collected from 19 young adult speakers of American English. In the production experiment, speakers repeated the words cod, cud, who'd, and hood in a carrier phrase at normal, clear, and fast rates. Articulatory movements and the associated acoustic signal were recorded, yielding measures of contrast distance between /a/ and /[see text for symbol]/ and between /u/ and /[see text for symbol]/. In the discrimination experiment, sets of seven natural-sounding stimuli ranging from cod to cud and who'd to hood were synthesized, based on productions by one male and one female speaker. The continua were then presented to each of the 19 speakers in labeling and discrimination tasks. Consistent with the hypothesis, speakers with discrimination scores above the median produced greater acoustic contrasts than speakers with discrimination scores at or below the median. Such a relation between speech production and perception is compatible with a model of speech production in which articulatory movements for vowels are planned primarily in auditory space. PMID- 15532665 TI - The control of aerodynamics, acoustics, and perceptual characteristics during speech production. AB - One of the most important areas of study in speech motor control is the identification of control variables, the variables controlled by the nervous system during motor tasks. The current study examined two hypotheses regarding control variables in speech production: (1) pressure and resistance in the vocal tract are controlled, and (2) perceptual and acoustic accuracy are controlled. Aerodynamic and acoustic data were collected on 20 subjects in three conditions, normally (NT), with an open air pressure bleed tube in place (TWB), and with a closed bleed tube in place (TNB). The voice recordings collected from the speakers in the production study were used in the perceptual study. Results showed that oral pressure (Po) was significantly lower in the TWB condition than in the NT and TNB conditions. The Po in the TWB condition seemed to be related to maintenance of subglottal pressure (Ps). Examination of the perceptual and acoustic data indicated that perceptual accuracy for [a] was achieved by maintaining Ps to preserve a steady sound pressure level, fundamental frequency, and voicing. Overall, it appeared speakers controlled pressure in compensating, but for the ultimate goal of maintaining acoustic and perceptual accuracy. PMID- 15532666 TI - A neural network model of the articulatory-acoustic forward mapping trained on recordings of articulatory parameters. AB - Three neural network models were trained on the forward mapping from articulatory positions to acoustic outputs for a single speaker of the Edinburgh multi-channel articulatory speech database. The model parameters (i.e., connection weights) were learned via the backpropagation of error signals generated by the difference between acoustic outputs of the models, and their acoustic targets. Efficacy of the trained models was assessed by subjecting the models' acoustic outputs to speech intelligibility tests. The results of these tests showed that enough phonetic information was captured by the models to support rates of word identification as high as 84%, approaching an identification rate of 92% for the actual target stimuli. These forward models could serve as one component of a data-driven articulatory synthesizer. The models also provide the first step toward building a model of spoken word acquisition and phonological development trained on real speech. PMID- 15532667 TI - Talker differences in clear and conversational speech: vowel intelligibility for normal-hearing listeners. AB - Several studies have shown that when a talker is instructed to speak as though talking to a hearing-impaired person, the resulting "clear" speech is significantly more intelligible than typical conversational speech. While variability among talkers during speech production is well known, only one study to date [Gagne et al., J. Acad. Rehab. Audiol. 27, 135-158 (1994)] has directly examined differences among talkers producing clear and conversational speech. Data from that study, which utilized ten talkers, suggested that talkers vary in the extent to which they improve their intelligibility by speaking clearly. Similar variability can be also seen in studies using smaller groups of talkers [e.g., Picheny, Durlach, and Braida, J. Speech Hear. Res. 28, 96-103 (1985)]. In the current paper, clear and conversational speech materials were recorded from 41 male and female talkers aged 18 to 45 years. A listening experiment demonstrated that for normal-hearing listeners in noise, vowel intelligibility varied widely among the 41 talkers for both speaking styles, as did the magnitude of the speaking style effect. While female talkers showed a larger clear speech vowel intelligibility benefit than male talkers, neither talker age nor prior experience communicating with hearing-impaired listeners significantly affected the speaking style effect. PMID- 15532668 TI - Clear speech perception in acoustic and electric hearing. AB - When instructed to speak clearly for people with hearing loss, a talker can effectively enhance the intelligibility of his/her speech by producing "clear" speech. We analyzed global acoustic properties of clear and conversational speech from two talkers and measured their speech intelligibility over a wide range of signal-to-noise ratios in acoustic and electric hearing. Consistent with previous studies, we found that clear speech had a slower overall rate, higher temporal amplitude modulations, and also produced higher intelligibility than conversational speech. To delineate the role of temporal amplitude modulations in clear speech, we extracted the temporal envelope from a number of frequency bands and replaced speech fine-structure with noise fine-structure to simulate cochlear implants. Although both simulated and actual cochlear-implant listeners required higher signal-to-noise ratios to achieve normal performance, a 3-4 dB difference in speech reception threshold was preserved between clear and conversational speech for all experimental conditions. These results suggest that while temporal fine structure is important for speech recognition in noise in general, the temporal envelope carries acoustic cues that contribute to the clear speech intelligibility advantage. PMID- 15532669 TI - Training native English speakers to perceive Japanese length contrasts in word versus sentence contexts. AB - This study investigated whether native speakers of American English with no knowledge of Japanese could learn to perceive Japanese vowel and consonant length distinctions through auditory training with immediate feedback. One group of participants was trained to identify the number of moras in Japanese words spoken in isolation (word training), and another group in sentences (sentence training). Trained groups' pretest and post-test scores in the words-in-isolation context (word context) and the words-in-sentences context (sentence context) were compared to those of an untrained control group. The questions addressed were whether there was an overall effect of training, and whether there were differential effects of two types of training. Both trained groups showed similar improvement in their overall test scores. The results suggested that learning in one context generalized to the other. However, an advantage of sentence training over word training was found: at the post-test, there was a greater difference between the scores of the two contexts for the word-training group than for the sentence-training group. The results are discussed in terms of the factors that might contribute to the differences in second language learning between the word and the sentence contexts. PMID- 15532670 TI - Development of a quick speech-in-noise test for measuring signal-to-noise ratio loss in normal-hearing and hearing-impaired listeners. AB - This paper describes a shortened and improved version of the Speech in Noise (SIN) Test (Etymotic Research, 1993). In the first two of four experiments, the level of a female talker relative to that of four-talker babble was adjusted sentence by sentence to produce 50% correct scores for normal-hearing subjects. In the second two experiments, those sentences-in-babble that produced either lack of equivalence or high across-subject variability in scores were discarded. These experiments produced 12 equivalent lists, each containing six sentences, with one sentence at each adjusted signal-to-noise ratio of 25, 20, 15, 10, 5, and 0 dB. Six additional lists were also made equivalent when the scores of particular pairs were averaged. The final lists comprise the "QuickSIN" test that measures the SNR a listener requires to understand 50% of key words in sentences in a background of babble. The standard deviation of single-list scores is 1.4 dB SNR for hearing-impaired subjects, based on test-retest data. A single QuickSIN list takes approximately one minute to administer and provides an estimate of SNR loss accurate to +/-2.7 dB at the 95% confidence level. PMID- 15532671 TI - Speech activity detection and enhancement of a moving speaker based on the wideband generalized likelihood ratio and microphone arrays. AB - The subject of this work is a unifying treatment of estimating the Direction of Arrival (DOA), detecting speech activity and suppressing noise in the case of a moving speaker by using a linear microphone array. The approach is based on the generalized likelihood ratio test applied to the framework of far-field, wideband moving sources (W-GLRT). It is shown that under certain distributional assumptions the W-GLRT provides a framework for the evaluation of DOA measurements against spurious DOAs, probabilistic speech activity detection as well as speech enhancement. As regards speech enhancement, we demonstrate the direct connection of W-GLRT with enhancement based on subspace methods. In addition, through the concept of directive a priori SNR we demonstrate its indirect connection with Minimum Mean Square Error spectral (MMSE_SA) and log spectral gain modification (MMSE_LSA). The efficiency of the approach is illustrated on a moving speaker when either additive white Gaussian or babble noise is present in the acoustical field at very low SNRs. PMID- 15532672 TI - Perfect harmony: a mathematical analysis of four historical tunings. AB - In Western music, a musical interval defined by the frequency ratio of two notes is generally considered consonant when the ratio is composed of small integers. Perfect harmony or an "ideal just scale," which has no exact solution, would require the division of an octave into 12 notes, each of which would be used to create six other consonant intervals. The purpose of this study is to analyze four well-known historical tunings to evaluate how well each one approximates perfect harmony. The analysis consists of a general evaluation in which all consonant intervals are given equal weighting and a specific evaluation for three preludes from Bach's "Well-Tempered Clavier," for which intervals are weighted in proportion to the duration of their occurrence. The four tunings, 5-limit just intonation, quarter-comma meantone temperament, well temperament (Werckmeister III), and equal temperament, are evaluated by measures of centrality, dispersion, distance, and dissonance. When all keys and consonant intervals are equally weighted, equal temperament demonstrates the strongest performance across a variety of measures, although it is not always the best tuning. Given C as the starting note for each tuning, equal temperament and well temperament perform strongly for the three "Well-Tempered Clavier" preludes examined. PMID- 15532673 TI - The dynamics and tuning of orchestral crotales. AB - An experimental and theoretical investigation of the acoustic and vibrational properties of orchestral crotales within the range C6 to C8 is reported. Interferograms of the acoustically important modes of vibration are presented and the frequencies are reported. It is shown that the acoustic spectra of crotales are not predicted by assuming that they are either thin circular plates or annular plates clamped at the center, despite the physical resemblance to these objects. Results from finite element analysis are presented that demonstrate how changing the size of the central mass affects the tuning of the instruments, and it is concluded that crotales are not currently designed to ensure optimal tuning. The possibility of using annular plates as crotales is also investigated and the physical parameters for such a set of instruments are presented. PMID- 15532674 TI - Vocal tract resonances in singing: the soprano voice. AB - The vocal tract resonances of trained soprano singers were measured while they sang a range of vowels softly at different pitches. The measurements were made by broad band acoustic excitation at the mouth, which allowed the resonances of the tract to be measured simultaneously with and independently from the harmonics of the voice. At low pitch, when the lowest resonance frequency R1 exceeded f0, the values of the first two resonances R1 and R2 varied little with frequency and had values consistent with normal speech. At higher pitches, however, when fo exceeded the value of R1 observed at low pitch, R1 increased with f0 so that R1 was approximately equal to f0. R2 also increased over this high pitch range, probably as an incidental consequence of the tuning of R1. R3 increased slightly but systematically, across the whole pitch range measured. There was no evidence that any resonances are tuned close to harmonics of the pitch frequency except for R1 at high pitch. The variations in R1 and R2 at high pitch mean that vowels move, converge, and overlap their positions on the vocal plane (R2,R1) to an extent that implies loss of intelligibility. PMID- 15532675 TI - Experimental validation of a tractable numerical model for focused ultrasound heating in flow-through tissue phantoms. AB - Heating from high intensity focused ultrasound (HIFU) can be used to control bleeding, both from individual blood vessels as well as from gross damage to the capillary bed. The presence of vascularity can limit one's ability to elevate the temperature owing to convective heat transport. In an effort to better understand the heating process in tissues with vascular structure we have developed a numerical simulation that couples models for ultrasound propagation, acoustic streaming, ultrasound heating and blood cooling in a Newtonian viscous medium. The 3-D simulation allows for the study of complicated biological structures and insonation geometries. We have also undertaken a series of in vitro experiments employing non-uniform flow-through tissue phantoms and designed to provide verification of the model predictions. We show that blood flow of 2 cm/s (6.4 ml/min through a 2.6 mm 'vessel') can reduce peak temperature in a vessel wall by 25%. We also show that HIFU intensities of 6.5 x 10(5) W/m2 can induce acoustic streaming with peak velocities up to 5 cm/s and this can reduce heating near a vessel wall by more than 10%. These results demonstrate that convective cooling is important in HIFU and can be accounted for within simulation models. PMID- 15532676 TI - Geographic variation and acoustic structure of the underwater vocalization of harbor seal (Phoca vitulina) in Norway, Sweden and Scotland. AB - The male harbor seal (Phoca vitulina) produces broadband nonharmonic vocalizations underwater during the breeding season. In total, 120 vocalizations from six colonies were analyzed to provide a description of the acoustic structure and for the presence of geographic variation. The complex harbor seal vocalizations may be described by how the frequency bandwidth varies over time. An algorithm that identifies the boundaries between noise and signal from digital spectrograms was developed in order to extract a frequency bandwidth contour. The contours were used as inputs for multivariate analysis. The vocalizations' sound types (e.g., pulsed sound, whistle, and broadband nonharmonic sound) were determined by comparing the vocalizations' spectrographic representations with sound waves produced by known sound sources. Comparison between colonies revealed differences in the frequency contours, as well as some geographical variation in use of sound types. The vocal differences may reflect a limited exchange of individuals between the six colonies due to long distances and strong site fidelity. Geographically different vocal repertoires have potential for identifying discrete breeding colonies of harbor seals, but more information is needed on the nature and extent of early movements of young, the degree of learning, and the stability of the vocal repertoire. A characteristic feature of many vocalizations in this study was the presence of tonal-like introductory phrases that fit into the categories pulsed sound and whistles. The functions of these phrases are unknown but may be important in distance perception and localization of the sound source. The potential behavioral consequences of the observed variability may be indicative of adaptations to different environmental properties influencing determination of distance and direction and plausible different male mating tactics. PMID- 15532677 TI - Biochemical evidence that an S431F mutation in acetylcholinesterase-1 of Aphis gossypii mediates resistance to pirimicarb and omethoate. AB - Molecular changes in acetylcholinesterase (AChE) leading to target-site resistance to carbamate and organophosphate insecticides have recently been identified. Of particular interest is the S431F mutation in ace2 and its orthologue ace1 of Myzus persicae Sulzer and Aphis gossypii Glover, respectively. This mutation has been correlated with resistance to pirimicarb, but biochemical evidence has not yet been provided. Here, we describe for the first time that recombinantly expressed AChE1 from A gossypii carrying the S431F mutation is insensitive to pirimicarb and omethoate, but sensitive to demeton-S-methyl and hypersensitive to carbofuran. Furthermore, site-directed mutagenesis of the serine residue at position 431 in ace1 from a pirimicarb-susceptible clone of A gossypii conferred insensitivity to pirimicarb. We conclude that AChE1 of A gossypii is the target of toxicological relevance of carbamates and organophosphates. PMID- 15532678 TI - In vivo distribution and metabolisation of 14C-imidacloprid in different compartments of Apis mellifera L. AB - In vivo distribution of the neonicotinoid insecticide, imidacloprid, was followed during 72 h in six biological compartments of Apis mellifera L: head, thorax, abdomen, haemolymph, midgut and rectum. Honeybees were treated orally with 100 microg of 14C-imidacloprid per kg of bee, a dose close to the median lethal dose. Elimination half-life of total radioactivity in honeybee was 25 h. Haemolymph was the compartment with the lowest and rectum that with the highest level of total radioactivity during the whole study, with a maximum 24h after treatment. Elimination half-life of imidacloprid in whole honeybee was 5 h. Imidacloprid was readily distributed and metabolised only by Phase I enzymes into five metabolites: 4/5-hydroxy-imidacloprid, 4,5-dihydroxy-imidacloprid, 6 chloronicotinic acid, and olefin and urea derivatives. The guanidine derivative was not detected. The urea derivative and 6-chloronicotinic acid were the main metabolites and appeared particularly in midgut and rectum. The olefin derivative and 4/5-hydroxy-imidacloprid preferentially occurred in head, thorax and abdomen, which are nicotinic acetylcholine receptor-rich tissues. Moreover, they presented a peak value around 4 h after imidacloprid ingestion. These results explain the prolongation of imidacloprid action in bees, and particularly the differences between rapid intoxication symptoms and late mortality. PMID- 15532679 TI - Synthesis and phloem mobility of acidic derivatives of the fungicide fenpiclonil. AB - A series of derivatives of the phenylpyrrole fungicide fenpiclonil was synthesized in which a carboxyl group was present at various sites of this non phloem-mobile molecule. Using the Kleier model, all these acidic analogues were predicted to be moderately phloem-mobile, especially the N-substituted derivatives. One of these latter molecules, N-carboxymethyl-3-cyano-4-(2,3 dichlorophenyl)pyrrole, exhibited some fungicidal activity on the pathogenic fungus Eutypa lata, and was then tested as a phloem-mobile pesticide in the Ricinus system. The compound was indeed mobile in the sieve tubes and was not degraded to fenpiclonil in the phloem sap under our experimental conditions. Its concentration in the sap was closely correlated to the percentage of the undissociated form of the molecule in the external medium, and was similar under acidic conditions (external pH 4.6-5.0) to that of the herbicide glyphosate. PMID- 15532680 TI - Delphastus catalinae and Coleomegilla maculata lengi (Coleoptera: Coccinellidae) as biological control agents of the greenhouse whitefly, trialeurodes vaporariorum (Homoptera: Aleyrodidae). AB - Predation efficacy and compatibility of the predatory lady beetles Coleomegilla maculata lengi Timberlake and Delphastus catalinae (Horn) against the greenhouse whitefly, Trialeurodes vaporariorum (Westwood) were studied in laboratory on glabrous fuchsia (Fuchsia hybrida Voss cv Lena Corolla) and pubescent poinsettia plants (Euphorbia pulcherrima Willd ex Klotzch cv Dark Red Annette Hegg). On glabrous plants (fuchsia), fourth-instar and adults of C maculata were the most efficient, both against whitefly eggs and pupae. On pubescent plants (poinsettia), the larger stages of C maculata were negatively affected and less efficient than adults of D catalinae. The presence of plant structure did not affect the voracity of either predator species. Finally, the simultaneous use of both predator species generated inter-specific competition. These results provide recommendations for biological control of whitefly in horticultural greenhouses. PMID- 15532681 TI - Photosynthetic and growth responses of zea mays L and four weed species following post-emergence treatments with mesotrione and atrazinet. AB - We compared photosynthesis and growth of Zea mays L (corn) and four weed species, Setaria viridis (L) Beauv (green foxtail), Echinochloa crus-galli (L) Beauv (barnyardgrass), Abutilon theophrasti Medic (velvetleaf), and Amaranthus retroflexus L (redroot pigweed), following foliar applications with atrazine, mesotrione, or a combination of atrazine and mesotrione in two greenhouse experiments. Plant responses to the three herbicide treatments were compared with responses of untreated plants (control). Photosynthesis on day 14 and dry mass of Z mays was not reduced by any of the herbicide treatments. Photosynthesis and dry mass of E crus-galli, A retroflexus and A theophrasti were significantly reduced by mesotrione and atrazine alone and in combination. Photosynthesis on day 14 and dry mass of large Sviridis plants were not suppressed by either herbicide applied alone. The mesotrione plus atrazine treatment was the most effective treatment for grass weed control because plants did not regain photosynthetic capacity and had significantly lower dry mass. Shoot dry mass of broadleaf weeds was significantly reduced by all three herbicide treatments, except for A retroflexus treated with mesotrione alone. PMID- 15532682 TI - Buffer zone and windbreak effects on spray drift deposition in a simulated wetland. AB - The amount of agricultural spray that drifts into a wetland from an adjacent crop field is influenced by vegetation along the field boundary or any intentional setback distance (buffer zone) between the sprayer and the edge of the arable field. In this study, spray tracer drift deposits were measured in a simulated wetland area under different conditions of wind speed and buffer zone width. The effect of an artificial windbreak at the upwind edge of the simulated wetland was also evaluated. A level of tolerance of 0.1% of the in-swath spray deposition was established as a no-effect level for the response of aquatic plants to common herbicides. Our results indicate that a vegetated 10-m field margin (eg a fencerow) alone provides adequate protection from herbicide drift into a wetland area under wind conditions normally considered acceptable for spraying. For high winds (> 4m s(-1)) when field spraying would not normally be advised, adequate protection was afforded by the same 10-m margin plus a dense windbreak (25% porosity) or by the margin plus a 20-m buffer zone. PMID- 15532683 TI - Activity of spinosad on stored-tobacco insects and persistence on cured tobacco stripst. AB - Every year raw tobacco and manufactured tobacco products are lost to two major storage pests, the cigarette beetle, Lasioderma serricorne (F) and the tobacco moth, Ephestia elutella (Hiibner). Post-harvest management of both insects is achieved through sanitation, insect monitoring and fumigation with phosphine. However, insect resistance to phosphine and control failures have been reported, and fumigants are under constant regulatory pressure. Here we report the evaluation of spinosad, a bioinsecticide derived from the fermentation of the soil micro-organism Saccharopolyspora spinosa Mertz & Yao. Spinosad was first registered in 1997 and is now widely used as a field pest control agent on many crops, including tobacco. The insecticidal activity of the fermentation product (technical spinosad, TS) was measured by diet incorporation assays against L serricorne and E elutella larvae. Mortality levels were determined on newly hatched larvae and over the whole insect life cycle. For both species, no emergence of adult insects was observed in cured tobacco sprayed with 50mg TS kg( 1) and inoculated with eggs or newly hatched larvae. These results indicated that spinosad has potential for the control of both species in stored tobacco, since 100% control of both pests could be achieved at 50 mg TS kg(-1), and with almost full control (90-95%) at 10 mg kg(-1). We also monitored the stability of the product on cured tobacco. The original concentration of the main active component of TS, spinosyn A, did not change significantly over 18 months, indicating no loss of spinosad during a typical leaf storage period of time. Bioassays against larvae confirmed that the bioinsecticidal activity of spinosad was retained. PMID- 15532684 TI - Per os efficacy of Ajuga extracts against sucking insects. AB - We studied the efficacy of water-soluble extracts from four Ajuga spp on the post embryonic development of two exopterygota (sucking insect) species. To allow comparison between different Ajuga species, results are expressed in terms of quantity of plant extracted per litre of test solution. Crude methanolic extracts of all Ajuga plants tested, with the exception of A genevensis, showed considerable per os efficacy against larvae of both Dysdercus cingulatus F and Acyrthosiphon pisum (Harris) even at 1 g litre(-1). In the aphid tests the order of efficacy was A bracteosa Wallich ex Benth > A chamaepitys Schreber > A reptans L > A genevensis L. On D cingulatus the order of efficacy was: A reptans > A bracteosa > A chamaepitys > A genevensis. Extracts were fractionated on SepPak using a range of methanol/water mixtures. Results are expressed in terms of the initial weight of plant extracted. The 100% methanolic fraction of A chamaepitys was highly effective on A pisum (100% mortality at 1 g litre(-1)) and less effective on D cingulatus (about 60% mortality at 5 g litre(-1)). The entire 60 methanol + 40 water fraction was effective against test insects but showed different efficacies according to test species and concentration applied. 20 Hydroxyecdysone (20E), cyasterone (Cy) and ajugalactone (Ajl) were identified in the fractions from all Ajuga species, but the remaining phytoecdysteroid profile was quite different between Ajuga species. Capitasterone (Cap) and 28-epi sengosterone (5Cy28') were found only in A reptans, makisterone A (MaA) and 29 norcyasterone (29NCy) were only in A chamaepitys, while 22-acetylcyasterone (Cy22A), 3-epi-cyasterone (Cy') and 3-epi-22-acetylcyasterone (Cy'22A) were only in A bracteosa. The total amount of phytoecdysteroids was 2053 mgkg(-1) for A bracteosa, 1892 mgkg(-1) for A reptans and 95 mg kg(-1) for A chamaepitys. PMID- 15532685 TI - Mechanisms of cuticular uptake of xenobiotics into living plants: 1. Influence of xenobiotic dose on the uptake of three model compounds applied in the absence and presence of surfactants into Chenopodium album, Hedera helix and Stephanotis floribunda leaves. AB - This study determined the uptake of three model compounds, applied in the presence and absence of surfactants, into the leaves of three plant species (Chenopodium album L, Hedera helix L and Stephanotis floribunda Brongn). The results with 2-deoxy-D-glucose, 2,4-dichlorophenoxyacetic acid and epoxiconazole in the presence ofsurfactants (the polyethylene glycol monododecyl ethers C12EO3, C12EO6, C12EO10 and a trisiloxane ethoxylate with mean EO of 7.5, all used at one equimolar concentration and therefore different percentage concentrations) illustrate that the initial dose (nmol mm(-2)) of xenobiotic applied to plant foliage is a strong positive determinant of uptake. This held true for all the xenobiotics studied over a wide concentration range in the presence of these surfactants. Uptake on a unit area basis (nmol mm(-2)) could be related to the initial dose of xenobiotic applied per unit area (ID) by an equation of the form: Uptake = a [ID]b at time t = 24h. ID is given by the mass of xenobiotic applied, M divided by the droplet spread area, A. Total mass uptake is then calculated from an equation of the form: Total Uptake = a [ID]b x A. PMID- 15532686 TI - Influence of phosphine on hatching of Cryptolestes ferrugineus (Coleoptera: Cucujidae), Lasioderma serricorne (Coleoptera: Anobiidae) and Oryzaephilus surinamensis (Coleoptera: Silvanidae). AB - The hatching and mortality response of 0- to 48-h-old eggs of field strains of the stored-product insects Cryptolestes ferrugineus (Stephens), Lasioderma serricorne (F) and Oryzaephilus surinamensis (L) following phosphine fumigation for 24, 48 or 120 h at 27 (+/- 2) degrees C was investigated. Hatching was delayed and reduced in the first few days in a phosphine-resistant strain of C ferrugineus that was treated with 2.0-7.0 mg litre(-1) doses for 48 h (5-80% mortality) and with 1.0-2.0 mg litre(-1) for 120 h (44-84% mortality). In both the exposures there were significant increases in hatching on later days when compared with the corresponding controls. Developmental delay was, however, not evident in susceptible strains of C ferrugineus, L serricorne and O surinamensis that were exposed to phosphine for 24 h. PMID- 15532687 TI - Synthesis and anti-mildew activity of 5-(2-aroyl)aryloxymethyl-2-phenyl-1 ,3,4 oxadiazoles against downy mildew of pearl millet. AB - A manipulatively simple, rapid, high-yielding and environmentally benign method for the integration of a heterocyclic ring, 1,3,4-oxadiazole, at the benzophenone nucleus has been achieved through intramolecular cyclization of substituted aroylaryloxyacetohydrazides to substituted 5-(2-aroyl)aryloxymethyl-2-phenyl 1,3,4-oxadiazoles under solventless 'dry' conditions using montmorillonite K10 clay and microwave irradiation. A comparison is made of the microwave-accelerated reaction with conventional heating conditions. Certain of the derivatives tested showed significant anti-mildew activity against Sclerospora graminicola (Sacc) Schroeter, the downy mildew pathogen of pearl millet. PMID- 15532688 TI - Repellency of aerosol and cream products containing fennel oil to mosquitoes under laboratory and field conditions. AB - The repellency of fennel (Foeniculum vulgare Miller)-containing products (5% aerosol and 8% cream) against mosquitoes was compared with those of citronella oil, geranium oil and deet, as well as three commercial repellents, Baby Keeper cream containing IR3535, MeiMei cream containing citronella and geranium oils, and Repellan S aerosol containing 19% N,N-diethyl-m-toluamide (deet) under laboratory and field conditions. In a laboratory study with female Aedes aegypti (L), fennel oil exhibited good repellency in a release-in-cage test and repellency in skin and patch tests of the oil was comparable with those of citronella and geranium oils. In paddy field tests with five human volunteers, 5% and 8% fennel oil-containing aerosol and cream produced 84% and 70% repellency, respectively, at 90 min after exposure, whereas Baby Keeper cream and MeiMei cream gave 71% and 57% repellency at 90 min after exposure, respectively, and Repellan S aerosol gave 89% repellency at 210 min. The species and ratio of mosquitoes collected were the genera Culex (44.1%), Anopheles (42.2%), Aedes (7.8%) and Armigeres (5.9%). Fennel oil-containing products could be useful for protection from humans and domestic animals from vector-borne diseases and nuisance caused by mosquitoes. PMID- 15532689 TI - Insecticidal activity of deoxypodophyllotoxin, isolated from Juniperus sabina L, and related lignans against larvae of Pieris rapae L. AB - In the course of screening for naturally occurring insecticides from plants from the northwestern part of China, a petroleum ether extract of Juniperus sabina L was found to show insecticidal activity against fifth-instar larvae of Pieris rapae L. From the extract, an insecticidal compound was isolated by bioassay guided fractionation. The compound was identified as deoxypodophyllotoxin (1) by comparison of its spectroscopic characteristics with literature data. In bioassays, 1 showed antifeedant activity against fifth-instar larvae of P rapae at 0.05-1.00 g litre(-1) and its AFC50 (concentration for 50% antifeedant activity) values at 12 and 48h were 0.170 and 0.060 g litre(-1), respectively. In that concentration range, all treated insects died within 48 h after treatment and compound 1 showed delayed insecticidal activity. At 0.015-0.100 g litre(-1), 1 showed insecticidal activity, with an LC50 of 0.020 g litre(-1). The related compound deoxypicropodophyllotoxin (2), however, showed lower antifeedant and insecticidal activities than 1 in bioassay. This indicated that the trans-lactone ring is an important moiety for enhancing activity in these compounds. Comparison of the insecticidal activities of 1 and another related compound, podophyllotoxin (3), suggested that varying the substituent at C-4 is an exciting possibility for synthesizing more potent analogues. PMID- 15532690 TI - Chemical residues and bioactivity of tebufenozide applied to apple foliage. AB - Tebufenozide, an insect growth regulator that acts as an ecdysone agonist, was applied at recommended label rate to apple trees in August 1997 and May 1998. Foliar residues were determined by high-performance liquid chromatography from samples collected 2, 24, 48 and 96 h post-spray, and at weekly intervals thereafter for 9 weeks in 1997 and 11 weeks in 1998. Foliage sampled at the same times was used in bioassays to determine residual toxicity to first-instar obliquebanded leafrollers, Choristoneura rosaceana (Harris). Residue decay followed first-order kinetics in both years, with residual half-life of 36.3 days in 1997 and 7.2 days in 1998. Estimates of the time needed to reduce bioactivity to 50% in bioassays were 18.7 days in 1997 and 36.3 days in 1998. The accuracy of equations describing decay of tebufenozide residues and bioactivity over time was not improved by using degree-day accumulations as the independent variable. PMID- 15532691 TI - Attractiveness of brown rice baits to non-target birds in harvested corn and soybean fields. AB - Brown rice is used as a bait-carrier for the avicide DRC-1339 (3-chloro-4 methylaniline) when baiting blackbirds (Icteridae). In March and April 1996 and 1997, we assessed non-target granivorous bird use of rice-baited plots placed in harvested corn and soybean fields in eastern South Dakota for 168 observation hours. In both years combined, we identified 10 and 14 granivorous species in corn and soybean fields, respectively. In 1996 (X = 0.2, SE = 0.04) and 1997 (X = 1.2, SE = 0.38), total numbers of granivores min(-1) were similar between crops (P > or = 0.322). In 1996, bird numbers were higher (P = 0.069) in rice-baited fields (X = 0.3, SE = 0.07) than in unbaited reference fields (X = 0.1, SE = 0.04). In 1997, bird numbers (X = 1.2, SE = 0.38) did not differ between treatments (P = 0.456). Our data show that small numbers of non-target birds visited the rice-baited plots. However, total number of different individual birds using the plots was unknown. PMID- 15532692 TI - Kinetics of interaction of three carbamate pesticides with Indian soils: Aligarh district. AB - The kinetics of interaction with soil of three carbamate pesticides (I, II, III) used as nematicides and herbicides was studied at four different temperatures from solutions of six soil samples of Aligarh district. The values obtained for rate constants for adsorption and desorption were in good agreement with those obtained from the Lindstrom model, which proved useful in the simultaneous evaluation of adsorption (k1) and desorption (k2) rate constants. The rate constants for pesticides were in the order III > I > II on all the six soil samples. The data for rate constants, activation energies, heats of activation, entropy of activation and thermodynamic parameters indicated a partly physical and partly chemical adsorption of pesticide on the soil surfaces. Adsorption occurred via coordination and/or protonation of the exchangeable cations with the amidic carbonyl group, and hydrogen bonding and dipole association at the crystal edge and basal surfaces. The adsorptivity of the soils may be attributed to the organic matter content and percentage clay content. PMID- 15532693 TI - Entangled, or tied in knots? PMID- 15532694 TI - Rapid induction of protective tolerance to potential terrorist agents: a systematic review of low- and ultra-low dose research. AB - OBJECTIVE: To systematically review the literature on the ability of low-dose (LD) and ultra-low-dose (ULD) toxin exposure to prevent and treat biological and chemical threats. METHODS: Laboratory research articles on protection or treatment from LD or ULD exposure for the 13 high-risk chemical and biological warfare threats were collected and systematically evaluated for quantity and scientific quality using pre-defined methodological criteria. RESULTS: Over 2600 articles were screened. Only five studies met the inclusion criteria examining stimulation and protective effects of LD- or ULD-exposures to the 13 pre identified biological and chemical agents. The quality evaluation (QE) of these studies was above average with a mean QE score of 70.6% of maximum. Two articles of fair to good quality reported both protective and treatment efficacy from exposure of animals or humans to LD- and ULD-exposures to toxins of risk in biochemical warfare. CONCLUSION: There is little research on agents of biological and chemical warfare investigating the possible use of LD- and ULD-toxins for protection and treatment. The existing literature is generally of good quality and indicates that rapid induction of protective tolerance is a feasible but under-investigated approach to bioterrorist or biowarfare defense. In our opinion, further research into the role of induced protection with LD- and ULD toxic agents is needed. PMID- 15532695 TI - Homeopathic proving symptoms: result of a local, non-local, or placebo process? A blinded, placebo-controlled pilot study. AB - BACKGROUND: Homeopathic pathogenetic trials (HPTs) (provings) are the pillar of homeopathy. Symptoms experienced by healthy volunteers are used to find the correct medicine for therapy. It is unclear whether these symptoms are specific or due to placebo noise. Furthermore, it is uncertain whether proving effects, if present at all, are due to a local or non-local process OBJECTIVES: To develop a test model which allows for testing if homeopathic proving symptoms are caused by placebo or causative mechanisms, and if these symptoms are due to local or non local processes. DESIGN: Randomised, blinded, placebo-controlled, parallel-group study, with 1-week baseline and 2-weeks proving period. SUBJECTS: 11 healthy volunteers from two different homeopathic schools. PROVING SUBSTANCE: An homeopathic medicine (Cantharis 30c), blindly chosen from 12 potential medicines, compared to placebo. OUTCOME MEASURE: Number of symptoms typical for the medicine in the experimental and control group during baseline and proving period. RESULTS: During baseline there was no difference in the number of typical or atypical symptoms in either group. During the proving period, both more typical symptoms for Cantharis (P= 0.03) and more atypical symptoms (P= 0.02) were observed compared to baseline. Between-group differences were not significant. Effect sizes for the difference between the proving and control group for typical symptoms was d=0.4, and for atypical symptoms d=0.6. DISCUSSION: This proving model could be valuable in studying the validity of proving symptoms of homeopathic substances in healthy volunteers. CONCLUSION: Homeopathic proving symptoms appear to be specific to the medicine and do not seem to be due to a local process. Since this was a pilot study using a small number of provers, rival hypotheses cannot be ruled out and the study needs replication. PMID- 15532696 TI - The placebo-controlled trial as a test of complementary and alternative medicine: observations from research experience of individualised homeopathic treatment. AB - The authors' experience of conducting clinical trials in homeopathy and analysing data from these has drawn attention to a fundamental problem with the interpretation of results from placebo controlled trials in homeopathy: It is not reasonable to assume that the specific effects of homeopathic medicine and the non-specific effects of consultations are independent of each other-specific effects of the medicine (as manifested by patients' reactions) may influence the nature of subsequent consultations and the non-specific effects of the consultation may enhance or diminish the effects of the medicine. For clinical trials of homeopathy to be accurate representations of practice, we need modified designs that take into account the complexity of the homeopathic intervention. Only with such trials will the results be generalisable to homeopathic practice in the real world. The authors propose that comparative trials are a meaningful way of evaluating the effectiveness of homeopathic treatment. PMID- 15532697 TI - Repertory and the symptom loquacity: some results from a pilot study on likelihood ratio. AB - Treatment outcome in a pilot study indicates that it is possible to assess likelihood ratios of homeopathic symptoms. Entries in repertory rubrics can be validated, but must still be handled carefully. Prospective research is the only acceptable way. Software to support this research must be carefully designed to export correct data. PMID- 15532698 TI - Effect of Atropa belladonna and Echinacea angustifolia in homeopathic dilution on experimental peritonitis. AB - Atropa belladonna and Echinacea angustifolia have been used in homeopathy as modulators of inflammatory processes, in simple potency or 'accord of potencies', as recommended by homotoxicology. We evaluated their effects on leukocyte migration and macrophage activity induced by experimental peritonitis in vivo. Mice were injected (i.p.) with LPS (1.0mg/kg) and treated (0.3ml/10g/day, s.c.) with different commercial forms of these medicines. Echinacea angustifolia D4--a simple potency preparation--and Belladonna Homaccord, Belladonna Injeel, Belladonna Injeel Forte, Echinacea Injeel and Echinacea Injeel Forte--all in 'accord of potencies'--were tested. The association of A. belladonna and E. angustifolia in 'accord of potencies' produced an increase of polymorphonuclear cell migration (Kruskal-Wallis, P = 0.03) and a decrease of mononuclear cell percentages (Kruskal-Wallis, P < or = 0.04), when compared with control, mainly in preparations containing low potencies. The proportion of degenerate leukocytes was lower in the treated groups, compared to a control group (P < or = 0.05). The treated groups showed increased phagocytosis (P < or = 0.05), mainly in preparations containing high potencies. Our results suggest that A. belladonna and E. angustifolia, when prepared in 'accord of potencies', modulate peritoneal inflammatory reaction and have a cytoprotective action on leukocytes. PMID- 15532699 TI - On the dynamics of water molecules at the protein solute interfaces. AB - Proteins, with the large variety of chemical groups they present at their molecular surface, are a class of molecules which can be very informative on most of the possible solute-solvent interactions. Hen egg white lysozyme has been used as a probe to investigate the complex solvent dynamics occurring at the protein surface, by analysing the results obtained from Nuclear Magnetic Resonance, X-ray diffractometry and Molecular Dynamics simulations. A consistent overall picture for the dynamics of water molecules close to the protein is obtained, suggesting that a rapid exchange occurs, in a picosecond timescale, among all the possible hydration surface sites both in solution and the solid state, excluding the possibility that solvent molecules can form liquid-crystal-like supramolecular adducts, which have been proposed as a molecular basis of 'memory of water'. PMID- 15532700 TI - A preliminary audit investigating remedy reactions including adverse events in routine homeopathic practice. AB - Homeopathic medicines are regarded as safe but practitioners report several types of healing or remedy reactions including aggravations, new symptoms and recurrence of old symptoms, some of which could be regarded as side effects or unwanted effects. Some remedy reactions may be regarded as adverse events. AUDIT QUESTIONS: Do such reactions occur within our unit, and if so, how frequently? Do patients regard these events as "adverse"? METHODS: The audit was carried out in the Bristol Homeopathic Hospital Outpatient Department. All patients were given a questionnaire to complete when at their first follow-up consultation approx 6-10 weeks after their first appointment. One hundred and sixteen patients were sampled over a 2-month period. RESULTS: Reactions were frequent: 28 out of the 116 (24%) patients, experienced an aggravation. Thirteen patients (11%) reported an adverse event even though 5 of those were patients who also reported an aggravation followed by an overall improvement of their symptoms. Thirty-one patients described new symptoms (27%) and 21(18%), a return of old symptoms. Those experiencing the latter appeared to have better outcomes. CONCLUSIONS: Remedy reactions are common in clinical practice; some patients experience them as adverse events. Systematically recording side effects would facilitate our understanding of these reactions and would enable standards to be set for audit of information and patient care. PMID- 15532701 TI - Homeopathic treatment of radiation-induced itching in breast cancer patients. A prospective observational study. AB - Following surgery for carcinoma of the breast, patients receive local radiotherapy. This can cause itching, which may be severe, in the radiation field. The affected skin usually is dry, rough and red. Twenty-five patients were treated homeopathically for radiation-induced itching. Fourteen patients developed itching during their course of post-operative radiation at 27 days median (range: 14-40). Eleven patients experienced itching in the radiation field after completion of treatment (median 21 days) after the end of their radiation treatment. A single dose of an individually selected homeopathic medicine in 30C dilution was given in the clinic, on the basis of repertorisation. Patients were asked to record a visual analogue scale (VAS) before prescription of the homeopathic medicine and at follow-up. Patients were evaluated at median 3 days (range: 1-27 days) after administration of the homeopathic medicine. In total, 14 of 25 patients (56%) responded to the first medicine. Nine patients had a second medicine, seven responded. Altogether 21 of 25 (84%) patients were successfully treated. The following medicines were employed successfully: Fl-ac 9/13, Rhus-t 3/5, Caust 2/3, Ign 2/2, Psor 2/2, gamma-ray 2/2 and Kali-bi 1/1. The VAS measurements before and after homeopathic treatment showed a reduction of the median value of 64mm (range: 20-100mm) to 34mm (median; range: 0-84mm). Homeopathic treatment of radiation-induced itching appears quite successful. The most frequently indicated and most frequently effective medicine was Fluoric acid. An approach that allows greater understanding of the patient as a whole in the short time available in a busy clinic may be required. PMID- 15532702 TI - The toxicology of Amanita virosa: the destroying angel. AB - This paper examines the biology and medical consequences of ingesting the potentially lethal poisonous mushroom, Amanita virosa, the Destroying Angel. The fungus, its structure, distribution and toxic components are described. Symptoms of human poisoning by A. virosa are described, following the order of Homeopathic Repertories. Laboratory values for comparison with normal values of haematology, biochemistry and urine analyses are given. PMID- 15532703 TI - 20 years ago: British Homoeopathic Journal, October 1984. PMID- 15532705 TI - Presence of Cx37 and lack of desmin in smooth muscle cells are early markers for arteriogenesis. AB - In search of early structural markers of arteriogenesis, we studied the expression of gap junction proteins as well as of contractile and cytoskeletal proteins in smooth muscle cells (SMCs) during coronary collateral vessel growth induced by chronic occlusion of the left circumflex artery (LCx) in the dog heart. We used confocal microscopy with antibodies against connexin37 (Cx37), alpha-smooth muscle actin (alpha-SM actin), calponin, desmin and vinculin. The quantitative confocal analysis of immunofluorescence intensity showed that (1) in normal vessels (NV), Cx37 was present in endothelium only, not in SMC. Calponin, alpha-SM actin, desmin and vinculin were evenly expressed in SMC. (2) In early growing V (EV) with minimal intima formation, alpha-SM actin, calponin and vinculin showed little change in SMC, but desmin was 3.3 times lower than in NV, and Cx37 was induced (NV 0 arbitrary units/microm2, EV 50.3). (3) In actively growing V (AV), alpha-SM actin, calponin and vinculin were 3-, 3.3- and 2.9-fold lower, respectively, in the neointima as compared to the media. However, Cx37 was 48.2 AU/microm2 in the media and 15.8 AU/microm2 in the neointima. Desmin was almost absent in the neointima and 5-fold reduced in the media. SMC, strongly positive for alpha-SM actin and calponin, expressed Cx37. Our findings indicate that induction of Cx37 and reduction of desmin precede the phenotypic changes of SMCs, which are characterized by down-regulation of alpha-SM actin, calponin and vinculin, and the formation of a neointima. An altered expression of Cx37 and desmin, therefore, are early markers for arteriogenesis in dog heart. PMID- 15532704 TI - Mitochondrial signaling pathways: a receiver/integrator organelle. AB - Research over the last decade has extended the prevailing view of cell mitochondrial function well beyond its critical bioenergetic role in supplying ATP. Recently, it has been recognized that the mitochondria play a critical role in cell regulatory and signaling events, in the responses of cells to a multiplicity of physiological and genetic stresses, inter-organelle communication, cell proliferation and cell death. Nevertheless, a broad-based review on mitochondrial signaling is not presently available. To bridge that gap, this review examines the perspective of mitochondria as the receiver integrator and transmitter of signals, dissecting the multiple and interrelated signaling pathways at both the molecular and biochemical levels with particular focus on nuclear and cytoplasmic factors, fundamentally involved in the shaping of the organelles' responses. We examine evidence that the mitochondria act as a dynamic receiver and integrator of numerous translocated signaling proteins (including protein kinases and transcription factors), regulatory Ca2+ fluxes and membrane phospholipids as well the transmission of mitochondrial-generated oxidative stress and energy-related signaling. Novel experimental approaches studying mitochondrial signaling including cell studies using metabolic inhibitors and genetic stresses (e.g. mtDNA depletion) are discussed. While there is abundant interest and information concerning its integral role to apoptosis, mitochondrial signaling also plays a fundamental role in proliferative pathways, nutrient sensing, inter-organellar cross-talk and in the responses of cells to metabolic transition and physiological stresses which remain relatively unexplored. PMID- 15532706 TI - Study of the PTEN gene expression and FAK phosphorylation in human hepatocarcinoma tissues and cell lines. AB - The tumor suppressor PTEN gene maps to chromosome 10q23.3 and encodes a dual specificity phosphatase. Mutations of this gene had been found in a variety of human tumors. In the present study, we analyzed the structure and expression of the PTEN gene in 34 hepatocellular carcinoma tissues and two hepatoma cell lines. We found neither homozygous nor hemizygous deletions in these samples. We, however, found point mutations in 4 of the 34 tissue samples. Five of ten hepatocellular carcinoma tissues showed reduced PTEN expression at mRNA level. HepG2 and SMMC-7721 hepatoma cells showed decreased PTEN expression at both mRNA and protein levels compared with immortalized L02 hepatic cells. PTEN mRNA in SMMC-7721 hepatoma cells could be reduced by TGF-betaI treatment. We also found that the phosphorylation levels of FAK in both of the hepatoma cell lines were higher than that in L02 hepatic cells. Transient expression of the PTEN gene in SMMC-7721 and HepG2 hepatoma cells resulted in decreased FAK phosphorylation. The level of FAK tyrosine phosphorylation appeared to be inversely correlated with the level of the PTEN protein. In summary, our results indicated that the function of the PTEN gene in hepatocarcinomas may be impaired mainly through point mutations and expression deficiency and that the defect of PTEN in tumor cells could alter the phosphorylation of FAK. PMID- 15532707 TI - Presence of normal creatine in the muscle of a patient with a mutation in the creatine transporter: a case study. AB - To date, more than seven families have been reported who carry a mutation in the X-linked creatine-transporter (CrT) gene. The resulting lack of creatine in the brain is associated with mental retardation, severe expressive language disorder, mild epilepsy, and a complete absence of Cr in the brain (measured using MRS). Conversely, these patients had no observable cardiac or musculo-skeletal deficits. In this case study, a 22-year-old patient underwent surgical repair for scoliosis. Proton MRS of this patient's brain demonstrated the near-absence of creatine and phosphocreatine within the cerebral white and deep gray matter structures. Cerebral atrophy was noted with serial MRI examinations. Subsequent genetic and metabolic analysis showed some biochemical anomalies consistent with a CrT deficiency. The mutation in this patient was identified as a deletion at phenylalanine 107 (delF107). Control muscle biopsies were obtained from archived samples, which had been taken with informed consent during routine muscle biopsies for diagnostic purposes. We determined that the total Cr concentration in the skeletal muscle biopsy was 39.3 +/- 2.94 mmol/kg wet wt., which is not significantly different from non-CrT controls, n = 3 (43.3 +/- 3.57 mmol/kg wet wt.). We conclude that the brain appears to lack the ability to transport creatine when there is a mutation in the CrT gene. However, the muscle utilizes another mechanism for maintaining normal creatine levels. Identifying this alternative creatine-transport mechanism may be useful in treating the neurologic and cognitive impairments of patients with creatine-transporter deficiency. PMID- 15532708 TI - Enhanced expression of cytosolic fatty acid binding protein and fatty acid uptake during liver regeneration in rats. AB - BACKGROUND/AIMS: Cytoplasmic liver fatty acid binding protein (L-FABP) has been suggested to be associated with cellular mitotic activity but the changes in L FABP mRNA and protein levels during liver regeneration following partial hepatectomy (PHx) are not clear. METHODS: In the present study, we determined the time course of L-FABP mRNA expression and L-FABP levels following 70% PHx using Northern and Western blot, respectively. To elucidate one of the roles for L-FABP in PHx, [3H]-palmitic acid clearance in hepatocytes isolated from 24 h post-PHx and control animals was assessed. RESULTS: L-FABP mRNA increased at 30 min, peaked at approximately 1 h (163 +/- 17%; mean +/- SE, n = 5), and returned to control levels 6 h post-PHx. L-FABP level also increased at 1 h but peaked at 24 h (219 +/- 41%; mean +/- SE, n = 5). Hepatocyte [3H]-palmitic acid clearance increased by 29% at 24-h post-PHx, suggesting an increased intracellular transport (or binding) function by L-FABP. Pre-treatment with dexamethasone statistically reduced L-FABP levels (29%) and suppressed the regenerative process (mitotic activity). CONCLUSIONS: L-FABP mRNA increased sharply in response to PHx but the increase was short lived, while L-FABP level increased at a later stage. Both L-FABP content and fatty acid uptake increased significantly during liver regeneration induced by PHx in rats. It is likely that L-FABP is one of the factors responsible for hepatic regeneration. PMID- 15532709 TI - Down-regulation of inducible nitric oxide synthase by lysophosphatidic acid in human respiratory epithelial cells. AB - Viral infection generally results in the activation of inducible nitric oxide synthase (iNOS or NOS2) in respiratory epithelial cells by inflammatory cytokines. Activated NOS2 catalyzes synthesis of nitric oxide (NO), which in excess can cause cellular injury. On the other hand, lysophosphatidic acid (LPA), a lipid mediator released from epithelial cells, platelets, and fibroblasts in injured tissue, functions in repair of cell injury. However, details of the mechanism for repair by LPA remain unknown. We demonstrated one effect of LPA favoring repair, specifically inhibition by LPA of cytokine-induced NOS2 protein and mRNA expression by human respiratory epithelial cells in vitro. NO production by LPA-treated, cytokine-stimulated cells was also reduced. These decreases were prevented by Rho kinase inhibition with Y-27632. Thus, down-regulation by LPA of cytokine-induced increases in NOS2 activity is likely to involve a Rho-dependent signaling pathway. Harmful biologic effects of NO in viral respiratory infection might be modified by therapeutic manipulations involving LPA or Rho. PMID- 15532710 TI - Critical segment of apocytochrome c for its insertion into membrane. AB - Apocytochrome c has a potent ability to insert spontaneously into membrane. To identify which sequences were critical for this insertion activity, a series of peptides N19, C8, C15 and C21, corresponding to sequences 1-19, 81-88, 74-88 and 68-88 of apocytochrome c, respectively, were synthesized and purified. Insertion ability into phospholipid monolayer, intrinsic fluorescence emission spectra, and the accessibility of peptide C21 to fluorescence quenchers: KI, acrylamide and HB showed that only segment 68-88 could insert into membrane, while other segments did not. CD spectra demonstrated that its interaction with liposomes containing negatively charged phospholipid could induce a partial alpha-helical conformation in peptide C21. It is interesting to note that a cooperation exists between segment 68-88 and 1-19 in the insertion of apocytochrome c and consequently translocation across membrane. PMID- 15532711 TI - N-terminal kinase, and c-Src are activated in human aortic smooth muscle cells by pressure stress. AB - Mechanical forces related to pressure and flow are important for cell hypertrophy and proliferation. There are still a few studies that examine responses of human vascular smooth muscle cells to pure pressure stress (transmural pressure). It is unclear as to which mechanisms are involved in cellular responses to pressure elevation. On the other hand, although the involvement of the local renin angiotensin system (RAS) in pressure-induced responses was reported, the results were contradictory. It still remains to be determined whether RAS in human vascular smooth muscle cells is activated by pure pressure stress. We studied the upstream signal transduction events of extracellular signal kinase (ERK) in response to atmospheric pressure stress and involvement of angiotensin II in pressure-induced cell proliferation in human aortic smooth muscle cells (HASMC). A pressure-loading apparatus was set up to examine the effects of atmospheric pressure on human aortic smooth muscle cells. Pressure application of 160 mmHg for 3 h produced cell proliferation and activated ERK and c-JUN N-terminal kinase (JNK). ACE inhibitor suppressed all of them. ERK kinase (MEK) inhibitor also suppressed cell proliferation stimulated by pure pressure. The phosphorylated c Src was increased by pure pressure stress. The treatment with c-Src kinase inhibitor suppressed pressure-induced proliferative response. In summary, our study found that ERK activation mediated pure pressure-induced proliferative response of HASMC. This activation was partly mediated by c-Src. PMID- 15532712 TI - Regulation of myotrophin gene by pressure overload and stretch. AB - Hemodynamic load is a major determinant of cardiac mass and its phenotype, but very little is known about how mechanical load is converted into intracellular signals of gene expression and regulation. We have shown earlier that factors other than blood pressure control play a role in the mechanism involved in the development or regression of myocardial hypertrophy. We have identified a soluble factor, myotrophin, from the hearts of spontaneously hypertensive rats and dilated cardiomyopathic humans, which stimulates protein synthesis both in neonatal and adult rat cardiac myocytes. Myotrophin gene has been mapped and shown to be a novel gene localized in human chromosome 7q-33. The present study was conducted to evaluate the mechanism by which myotrophin is released and in turn initiates myocardial hypertrophy. We used an in vitro model, where neonatal cardiac myocytes were grown on stretchable plates and examined the effect of stretch on myotrophin gene expression (to mimic pressure overload), an in vivo model using beating non-working hearts exposed to high pressure and three different models of hypertensive rats. Our data showed that both cyclic stretch and exposure to high pressure caused significant increase in the transcript levels of myotrophin followed by expression of beta-myosin heavy chain and atrial natriuretic factor associated with an increase in myocardial protein synthesis. All three models of hypertensive rats also showed a significant increase in myotrophin transcripts. Altogether, our data strongly suggest that stretching of the cells by pressure or volume turns on the myotrophin, which in turn is responsible for the initiation process of myocardial hypertrophy in response to pressure or volume overload. PMID- 15532713 TI - Effect of angiotensin II on energetics, glucose metabolism and cytosolic NADH/NAD and NADPH/NADP redox in vascular smooth muscle. AB - Angiotensin II (AII) is a neurohormone and contractile agonist of vascular smooth muscle that has been shown to be involved in the pathogenesis of vascular disease, which may be partially caused by its effect on oxidant stress. Energy metabolism was examined in pig carotid arteries treated with AII, because the activity of pathways of intermediary metabolism of glucose determines the status of cytosolic NADH/NAD and NADPH/NADP redox, factors which are involved in oxidant stress. Contractile responses to AII were characterized by an increase in isometric force followed by a gradual decline to near-basal levels. Despite contractile activation, no change in glycolysis, lactate production, glucose oxidation, fatty acid oxidation, O2 consumption, glycogen content or high-energy phosphates was detected when compared to resting unstimulated arteries. Paradoxically, total uptake of glucose was inhibited by AII. Treatment with diphenylene iodinium, an inhibitor of NAD(P)H oxidase and superoxide production, reversed the inhibition of glucose uptake and revealed the expected increase in glucose uptake and oxidation upon contractile activation of smooth muscle by AII. The intracellular [lactate]/[pyruvate] ratio was increased, reflecting an increase in cytosolic NADH/NAD redox, whereas NADPH/NADP redox was decreased by AII. No change in NADPH/NADP redox was observed when membrane depolarization with K+ was used as the contractile agent. It is concluded that the pattern of force generation, metabolism and energetics of AII-stimulated contraction are significantly different from that of other contractile agonists. Most notably AII inhibited glucose uptake. NAD(P)H oxidase and/or attendant superoxide may play a role in modulating glucose metabolism. AII induces opposite changes in NADH/NAD redox and NADPH/NADP redox, which may have important consequences for oxidant stress. PMID- 15532714 TI - 17-Beta estradiol enhances prostaglandin E2 production in human U937-derived macrophages. AB - Prostaglandins (PGs) are potent eicosanoid lipid mediators that have been implicated in numerous homeostatic functions and inflammation. Estrogens have been shown to regulate the expression of genes in lipid metabolism in many cellular systems. In this study, the activation of macrophages and the modulation of PG release by estrogens were examined. The effects of 17-alpha and 17-beta estradiols, phytoestrogen Genistein and several selective estrogen receptor modulators on the release of PGE2 were investigated in human U937-derived macrophages. 17-Beta estradiol caused an enhancement of PGE2 production in a time and dose-dependent manner. Treatment of macrophages with 17-beta estradiol elicited an increased arachidonic acid (AA) release and an up-regulation of both cyclooxygenesis-1 and cyclooxygenesis-2 enzymes at both the transcript and protein levels. In addition, immunostaining of nuclear estrogen receptor alpha and the observation of ICI182 780 blockade of PGE2 production indicated that 17 beta estradiol-induced PGE2 release was mainly through nuclear estrogen receptor alpha. PMID- 15532715 TI - Activation and inactivation of volume-sensitive taurine efflux from rat mammary gland. AB - A knowledge of volume-sensitive solute transport in mammary cells is important in light of evidence that mammary cell metabolism is regulated by the cellular hydration state. In this report we have examined volume-sensitive taurine and K+ (Rb+) transport by lactating rat mammary tissue. A hyposmotic shock increased taurine efflux from rat mammary tissue: taurine release returned to a basal level upon transferring the tissue back to an isosmotic medium. However, the time taken to activate taurine efflux was less than the time taken to inactivate taurine release. A second subsequent osmotic challenge also increased taurine release but to a lesser extent than the first osmotic shock. A similar pattern was observed for bumetanide-insensitive, volume-activated K+ (Rb+) release from mammary tissue explants suggesting that taurine and K+ efflux are acting in concert to regulate mammary cell volume. An abrupt hyposmotic shock increased taurine efflux from mammary explants to a greater extent than a gradual reduction in the osmolality of the incubation medium. Increasing extracellular K+ increased taurine release via a pathway sensitive to niflumic acid, which suggests that activation of volume-sensitive taurine efflux does not require a change in the ionic strength of the incubation medium or a decrease in intracellular osmolality. A hyposmotic shock also stimulated taurine efflux from rat mammary acini. In contrast, a hyposmotic challenge had no effect on taurine uptake measured under sodium-free conditions. Hyposmotically induced taurine efflux was not dependent upon extracellular calcium. The results suggest that taurine and K+ transport may allow mammary cells to volume-regulate and consequently help to control mammary metabolism. PMID- 15532716 TI - AMP-deaminase from normal and cirrhotic human liver: a comparative study. AB - AMP-deaminase (EC 3.5.4.6) is an enzyme of nucleotide breakdown involved in regulation of adenine nucleotide pool in mammalian cells. Reaction catalysed by AMP-deaminase constitutes a rate-limiting step in adenine nucleotide catabolism in liver. In this study kinetic and regulatory properties of AMP-deaminase purified from normal and cirrhotic human liver were investigated. In comparison to AMP-deaminase extracted from the normal human liver, AMP-deaminase extracted from the cirrhotic liver was less sensitive towards substrate analogues, and only a very limited response towards pH and adenylate energy charge changes tested for enzyme isolated from this tissue source had been observed. At physiological pH 7.0, in the absence and in the presence of important allosteric effectors (ATP, ADP, GTP and orthophosphate), AMP-deaminases from the two sources studied manifested different regulatory profiles, with half-saturation constant (S0.5) values being distinctly higher for the enzyme extracted from the pathological organ. In contrast to AMP-deaminase isolated from the normal, healthy liver, where presence of relatively large (68 kDa) protein fragment was also detected, only smaller protein fragments were identified, while SDS-PAG electrophoresis of AMP-deaminase isolated from the cirrhotic liver was performed. The obtained results indicate clearly that advanced proteolytic processes occurring in the cirrhotic liver may affect structural integrity of AMP-deaminase studied, making enzyme less active and less sensitive to regulatory action of important allosteric effectors. PMID- 15532717 TI - Hydroxyl radical scavenging and singlet oxygen quenching properties of polyamines. AB - Polyamines (cadaverine, putrescine, spermidine, spermine) have been shown to be present in all prokaryotic and eukaryotic cells, and proposed to be important anti-inflammatory agents. Some polyamines at high concentrations are known to scavenge superoxide radicals in vitro. We have investigated the possible antioxidant properties of polyamines and found that polyamines, e.g., cadaverine, putrescine, spermidine and spermine do not scavenge superoxide radicals at 0.5, 1.0 and 2 mM concentrations. However, polyamines were found to be potent scavengers of hydroxyl radicals. Hydroxyl radicals were produced in a Fenton type reaction and detected as DMPO-OH adducts by electron paramagnetic resonance spectroscopic technique. Spermine, spermidine, putrescine and cadaverine inhibited DMPO-OH adduct formation in a dose dependent manner, and at 1.5 mM concentration virtually eliminated the adduct formation. The *OH-dependent TBA reactive product of deoxyribose was also inhibited by polyamines in a dose dependent manner. Polyamines were also found to inhibit the 1O2-dependent 2,2,6,6 tetramethylpiperidine N-oxy 1 (TEMPO) formation. 1O2 was produced in a photosensitizing system using Rose Bengal or Methylene Blue as photosensitizers, and was detected as TEMP-1O2 adduct by EPR spectroscopy. Spermine or spermidine inhibited the 1O2-dependent TEMPO formation maximally to 50%, whereas putrescine or cadaverine inhibited this reaction only up to 15%, when used at 0.5 and 1 mM concentrations. These results suggest that polyamines are powerful. OH scavengers, and spermine or spermidine also can quench singlet oxygen at higher concentrations. PMID- 15532719 TI - Altered Mg2+ transport across liver plasma membrane from streptozotocin-treated rats. AB - Type-I diabetes is associated with a decrease in magnesium content in various tissues, including liver. We have reported that hepatocytes from streptozotocin injected rats have lost the ability to accumulate Mg2+ following hormonal stimulation. To assess whether the defect is inherent to the Mg2+ transport mechanism located in the hepatocyte cell membrane, plasma membrane vesicles were purified from diabetic livers. Diabetic plasma membranes do not retain intravesicular Mg2+ as tightly as vesicles purified from livers of age-matched non-diabetic rats. In addition, the amount of intravesicular Mg2+ these vesicles exchange for extravesicular Na+ or Ca2+ is 2-3-fold larger than in non-diabetic vesicles. The partition of Ca2+/Mg2+ and Na+/Mg2+ exchange mechanisms in the apical and basolateral domains of liver plasma membrane is maintained under diabetic conditions, although the Na+/Mg2+ exchanger in diabetic basolateral membranes has lost the ability to operate in reverse and favor an accumulation of extravesicular Mg2+ within the vesicles in exchange for entrapped Na+. These data indicate the occurrence of a major alteration in Mg2+ transport across the hepatocyte membrane, which can explain, at least in part, the decrease in liver magnesium content observed in diabetic animals and patients. PMID- 15532718 TI - Regulation of protein kinase C isozymes in volume overload cardiac hypertrophy. AB - Protein kinase C (PKC) is a family of at least 11 isozymes and known to play a crucial role in myocardial growth. The present study was performed to investigate whether PKC-isozymes are differentially regulated during the development of volume-overload cardiac hypertrophy. After 2, 7 and 30 days of sham or aortocaval shunt operation in male Wistar rats, PKC-activity and the expression of cardiac PKC-isozymes (PKC-alpha, delta and epsilon) were determined at the protein and at the mRNA-level in the left and the right ventricle separately. Myocardial hypertrophy after 2, 7 and 30 days of aortocaval shunt was more pronounced in the right than in the left ventricle. Right ventricular hypertrophy was associated with an increased PKC-enzyme activity, a selectively enhanced protein expression of cytosolic PKC-delta (day 7: +83 +/- 12%, day 30: +94 +/- 14%) and PKC-alpha (day 7: +48 +/- 11%, day 30: +62 +/- 16%) and a transcriptional upregulation of the absolute mRNA-levels of these PKC-isozymes in the aortocaval shunt group as compared to controls. In contrast, the expression of PKC-epsilon was unchanged. A significant upregulation of PKC-delta both on the protein and on the mRNA-level was also noted in volume-overload induced left ventricular hypertrophy, whereas the expression of PKC-alpha and PKC-epsilon were not altered. Furthermore, the expression of calcineurin in both ventricles was not significantly changed in response to volume-overload. This study characterizes in the left and right ventricle a differential regulation of the dominant PKC-isozymes in volume overload cardiac hypertrophy both at the protein and at the mRNA-level. PMID- 15532720 TI - Diabetic cardiomyocyte dysfunction and myocyte insulin resistance: role of glucose-induced PKC activity. AB - Increased protein kinase C (PKC) activity has been implicated in the pathogenesis of a number of diabetic complications, and high concentrations of glucose have been shown to increase PKC activity. The present study was designed to examine the role of PKC in diabetes-induced (and glucose-induced) cardiomyocyte dysfunction and insulin resistance (measured by glucose uptake). Adult rat ventricular myocytes were isolated from nondiabetic and type 1 diabetic animals (4-5 days post-streptozotocin treatment), and maintained overnight, with/without the nonspecific PKC inhibitor chelerythrine (CHEL = 1 microM). Myocyte mechanical properties were evaluated using a video edge-detection system. Basal and insulin stimulated glucose uptake was measured with [3H]-2-deoxyglucose. Blunted insulin stimulated glucose uptake was apparent in diabetic myocytes, and both mechanical dysfunctions (e.g., slowed shortening/relengthening) and insulin resistance were maintained in culture, and normalized by CHEL. Cardiomyocytes isolated from nondiabetic animals were cultured in a high concentration of glucose (HG = 25.5 mM) medium, with/without CHEL. HG myocytes exhibited slowed shortening/relengthening and impaired insulin-stimulated glucose uptake compared to myocytes cultured in normal glucose (5.5 mM), and both impairments were prevented by culturing cells in CHEL. Our data support the view that PKC activation contributes to both diabetes-induced abnormal cardiomyocyte mechanics and insulin resistance, and that elevated glucose is sufficient to induce these effects. PMID- 15532721 TI - Adenovirus mediated overexpression of CYP2E1 increases sensitivity of HepG2 cells to acetaminophen induced cytotoxicity. AB - To study the biochemical and toxicological properties of cytochrome P450 2E1 (CYP2E1), an adenovirus containing human CYP2E1 cDNA (Ad-CYP2E1) was constructed and was shown to successfully mediate the overexpression of CYP2E1 in HepG2 cells. Acetaminophen (APAP) toxicity to HepG2 cells infected with Ad-CYP2E1 was characterized as a preliminary proof of principle experiment to validate the functionality of the CYP2E1 adenovirus. Compared with cells infected with Ad LacZ, HepG2 cells infected with Ad-CYP2E1 were more sensitive to APAP induced necrosis and apoptosis when the cells were depleted of intracellular reduced glutathione (GSH). The APAP cytotoxicity was dependent on both the concentration of APAP and the multiplicity of infection of the Ad-CYP2E1 virus. Apoptosis induced by APAP in HepG2 cells overexpressing CYP2E1 was caspase dependent and could be inhibited by the pan-caspase inhibitor Z-VAD-fmk. After treatment with APAP, mitochondrial membrane potential was dramatically decreased in the CYP2E1 expressing cells. APAP protein adducts were elevated in HepG2 cells infected with Ad-CYP2E1 compared with that in cells infected with Ad-LacZ; two bands around 90 KD were found only in the CYP2E1-expressing cells. These results demonstrate that adenovirus-mediated overexpression of human CYP2E1 activates APAP to reactive metabolites which damage mitochondria, form protein adducts, and result in toxicity to HepG2 cells. The Ad-CYP2E1 may be useful for studies designed to investigate the role of CYP2E1 in APAP and alcoholic liver injury and to further characterize the actions and effects of CYP2E1. PMID- 15532722 TI - Inhibition of nuclear factor-kappa B prevents staphylococcal enterotoxin A induced fever. AB - It has been shown that staphylococcal enterotoxin A (SEA) acts through human peripheral blood mononuclear cells (PBMC) to stimulate synthesis or release of pyrogenic cytokines. Nuclear factor-kappa B (NF-kappaB) is thought to play an important role in inflammatory responses through the regulation of genes encoding pro-inflammatory cytokines. The purpose of the present study was to determine whether the NF-kappaB mechanisms in human PBMC are involved in SEA-induced fever. Western blot evaluation revealed SEA was able to induce nuclear translocation of NF-kappaB from cytosol to nucleus in PBMC, which could be abolished by a NF kappaB inhibitor such as pyrrolidine dithiocarbamate (PDTC), sodium pyrithione (Pyri), N-acetyl-L-cysteine (NAC), or curcumin (Cur). Electrophoretic mobility shift assay also showed that the NF-kappaB DNA-binding activity was increased in the SEA-treated PBMC. Again, the SEA-induced increased NF-kappaB binding activity was significantly attenuated by either PDTC, Pyri, NAC or Cur. The pyrogenic responses to supernatant fluids obtained from human PBMC stimulated with SEA were associated with increased levels of interleukin 1-beta (IL-1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) in the supernatant fluids. Both the fever and the increased levels of IL-1beta, IL-6, and TNF-alpha in supernatant fluids obtained from the SEA-stimulated PBMC were decreased by incubating SEA-PBMC with either PDTC, Pyri, NAC, or Cur. Furthermore, the fever induced by systemic or central administration of SEA in rabbits were attenuated by pre-treatment with an systemic or central dose of either PDTC, Pyri, NAC, or Cur. The data indicate that inhibition of NF-kappaB prevents SEA-induced fever. PMID- 15532723 TI - Ribosomal protein S18 identified as a cofilin-binding protein by using phage display library. AB - We previously reported that an actin-binding protein, cofilin, is involved in superoxide production, phagocytosis, and chemotaxis in activated phagocytes through cytoskeletal reorganization. To elucidate the functions of cofilin in greater detail we tried to identify cofilin-binding proteins by using a phage displayed cDNA library constructed from human brain mRNAs. Several phage clones capable of binding to cofilin were obtained, and the phage with the strongest binding affinity contained the C-terminal half of ribosomal protein S18. To confirm the interaction between the S18 protein and cofilin, we investigated whether cofilin would bind to His-tagged S18 protein immobilized in Ni-NTA agarose gel. Cofilin and the S18 protein co-eluted with a low pH (4.5) buffer, suggesting that the proteins interact with each other. Preincubation of cofilin with actin abrogated the binding to protein S18, indicating that cofilin interacts with S18 protein at the actin-binding site, and cofilin co immunoprecipitated with FLAG-tagged S18 protein expressed in COS-7 cells. These results suggest that some cofilin molecules bind the ribosomal S18 protein under physiological conditions. PMID- 15532724 TI - Sesame lignans enhance antioxidant activity of vitamin E in lipid peroxidation systems. AB - The antioxidant properties of sesame lignans (sesamol, sesamin and sesamolin) were evaluated in comparison to tocols (alpha- and gamma-tocopherols and alpha tocotrienol) and butylated hydroxytoluene (BHT) using the following in vitro lipid peroxidation systems: (i) rat liver microsomes and cumene hydroperoxide (CumOOH)/Fe2+-ADP-NADPH (enzymatic) or (ii) rat liver mitochondria and Fe2+ ascorbate (nonenzymatic) systems. Sesamol containing a free phenolic group inhibited lipid peroxidation in both the systems whereas sesamin and sesamolin having methylenedioxy groups were effective only in the microsomal system. Since detoxifying enzymes are localized in microsomes, the inhibitory effects of sesamin and sesamolin observed in the microsomal system may be attributed to their metabolites. However, the inhibitory effects of lignans were lower than tocols and BHT. Combination of individual lignans and tocopherols (alpha, gamma) or alpha-tocotrienol showed higher inhibitory effects than the sum of individual inhibitions in CumOOH and Fe2+-ascorbate systems suggesting synergistic interactions. The time course of CumOOH-mediated lipid peroxidation showed a lag period and a decreased rate of thiobarbituric acid reactive product formation in the presence of individual lignans in combination with alpha-tocopherol suggesting recycling of alpha-tocopherol. PMID- 15532725 TI - AT2 receptor and apoptosis during AT1 receptor blockade in reperfused myocardial infarction in the rat. AB - We assessed whether upregulation of the angiotensin II (AngII) type 2 receptor (AT2R) during AngII type 1 receptor (AT1R) blockade might induce apoptosis in the in vivo rat model of reperfused myocardial infarction (RMI) and whether addition of an AT2R blocker abolishes that effect. We measured in vivo hemodynamics and left ventricular (LV) systolic and diastolic function (echocardiograms/Doppler), and ex vivo infarct size (triphenyl tetrazolium chloride), regional AT1R and AT2R proteins (immunoblots), and apoptosis (TUNEL assay and DNA ladder) after regional anterior RMI (60 min ischemia, 90 min reperfusion) in Sprague-Dawley rats randomized to intravenous AT1R blockade with candesartan (1 mg/kg, n = 9) or saline (controls, n = 14) over 30 min before RMI, and sham (n = 8). We also assessed the effect of AT2R blockade (PD123319, 10 mg/kg i.v.) plus candesartan on infarct size and apoptosis. Compared to controls, candesartan significantly (p < 0.001) limited increases in left atrial pressure, improved positive LV dP/dtmax and negative dP/dtmin, normalized LV ejection fraction, improved LV diastolic function, limited infarct expansion, decreased infarct size and apoptosis, and increased AT2R protein (not AT1R) in the reperfused ischemic zone. There were no changes in sham hearts. PD123319 abolished the candesartan-induced decrease in infarct size and LV dysfunction but not the decrease in apoptosis. Thus, during AT1R blockade in the in vivo rat model of RMI, regional AT2R upregulation contributes to the beneficial effect on infarct size and LV dysfunction but not on apoptosis, suggesting that the apoptosis is AT1R not AT2R-mediated. PMID- 15532726 TI - Differential expression of VEGF-A mRNA by 17beta-estradiol in breast tumor cells lacking classical ER-alpha may be mediated through a variant form of ER-alpha. AB - Beta-estradiol (17beta-E2) augments VEGF-A expression in various estrogen targeted organs and cells including breast tumor derived cell lines, via an ER alpha mediated pathway. Ironically, 17beta-E2 is able to regulate some genes via ER-alpha independent pathways. In the present study, we sought to determine whether 17beta-E2 can modulate VEGF-A expression in absence of ER-alpha, and therefore, three different cell lines including ER-alpha+ MCF-7, and ER-alpha SKBR-3 and HMEC were used for this study. The present study demonstrates that 17beta-E2 also induces VEGF-A mRNA expression in ER-negative SKBR-3 breast tumor cells in a manner similar to that observed in ER-positive MCF-7 cells. Blocking the induced-expression by antiestrogen ICI 182,780 indicates the induction pathway is ER dependent. While ER-alpha mRNA is absent in both HMEC and SKBR-3 cells, the impact of estrogen was found only in SKBR-3 cells, suggesting the existence of an analogue to ER-alpha or overlapping signal in these cells. Consistent with this suggestion, the present studies demonstrate the existence of an ER-alpha(var2) protein in MCF-7 and in SKBR-3 cells. This variant is predominantly localized in the nuclei of SKBR-3 cells. Importantly, specific binding of 17beta-E2 by these cells suggest the ER-alpha(var2) may act as active receptor in SKBR-3 cells. PMID- 15532727 TI - Cloning and characterization of chironomidae ferrochelatase: copper activation of the purified ferrochelatase. AB - All organisms utilize ferrochelatase (EC 4.99.1.1) to catalyze the insertion of ferrous iron into protoposphyrin IX in the terminal step of the heme biosynthetic pathway. Different metal-binding affinity for the enzyme leads to changes in enzyme activity. In this work, we have cloned and over-expressed the enzyme from chironomidae in E. coli. The enzyme was purified and characterized. The recombinant enzyme showed higher enzymatic activity (four-fold increase) in the presence of copper ions and unaffected by calcium ions. Other divalent metal ions including magnesium, manganese, lead, reduced the enzyme activity by >60%. Over 90% of the enzyme activity was inhibited by Zn2+. The sequence alignment of amino acid residues reveals 83% homology with other ferrochelatases. The results of electron proton resonance (EPR) analysis showed that Fe2+ ion was present in the cluster of the recombinant enzyme complex. The recombinant enzyme also contained the [2Fe-2S] center with two-fold higher enzymatic activity than human ferrochelatase. PMID- 15532729 TI - [The most frequent kinds of vertigo and their therapy: controlling the merry-go round in the head of patients]. PMID- 15532728 TI - Morphogenetic and apoptotic changes in diabetic cataract: prevention by pyruvate. AB - Studies have been conducted to ascertain the preventive effect of pyruvate against diabetes induced damage to DNA and associated morphogenetic changes in the mouse lens. Such changes were characterized by DNA nicks as well as by gross morphological changes in the nuclei, evident respectively by TUNEL and Hoechst staining procedures. Morphogenetic changes were also apparent by abnormal diferentiation of the germinal epithelial cells and errors in their migratory pathway. These changes were prevented by simultaneous administration of pyruvate to the diabetic animals. The preventive effect of this agent is attributable to its property of scavenging oxy-radicals generated by high levels of the sugars. PMID- 15532730 TI - [Calming most patients with vertigo. The course is usually benign--therapy successful (interview by Dr. Judith Neumaier)]. PMID- 15532731 TI - [Asymptomatic carotid stenoses: is surgery performed too quickly? (interview by Dr. Thomas Meissner)]. PMID- 15532732 TI - [Increased temperature and malfunctioning liver: what is the etiology? Tropical fever in family practice]. PMID- 15532733 TI - [A consequence of the obesity epidemic--the life expectancy of our children is on the decline]. PMID- 15532734 TI - [Treatment of obese children and adolescents--necessity and options]. AB - The alarming increase in obesity in children and adolescents has prompted physicians, dietary scientists, psychologists and PT instructors to concern themselves with the therapeutic possibilities available, and to implement them. An interdisciplinary approach, the scope and time frame of which is orientated to the degree of obesity has proved promising. By educating parents, multimodal programs support attempts to change family eating and exercise habits in the direction of a healthier lifestyle. The interlinking of various institutions ranging from general practitioners to rehabilitation centers on the basis of a common documentation structure with the aim of offering evidence-based treatment over the long term would be desirable. PMID- 15532736 TI - [Pharmacotherapy of obesity]. AB - While the intentional modification of eating habits remains the most important measure for losing weight, pharmacological support, in particular with regard to the long-term outcome, can be helpful. Catacholaminergic substances, such as diethylpropion and phenylpropanolamine are restricted by their side effects to short-term application only, and their use makes little sense. The serotoninergic substance sibutramine, which acts on the central nervous system to curb the appetite or enhance the feeling of saturation, and orlistat, which inhibits the assimilation of fat from the bowel, are approved for longer-term treatment of obesity, and lead to an additional loss of 3-5% of the patient's initial weight after one year of use. Thereafter, however, this benefit decreases. Further substances involving new, possibly combinable, approaches are currently under development. PMID- 15532735 TI - [Low-carbohydrate or low-fat diet for weight loss--which is better?]. AB - Several recent clinical studies show that a low-carbohydrate diet produces a greater initial weight loss than conventional low-fat diets, and is associated with a greater reduction of elevated serum triglycerides. After one year, however, weight loss is similar with both diets. Since the intake of saturated fat is higher on a low-carbohydrate diet, there may be an increased risk of elevated levels of LDL cholesterol, thus furthering atherosclerosis, over the long term. Before low-carbohydrate diets can be considered an equivalent alternative to low-fat diets for the treatment of obesity, long-term clinical trials are urgently required. The greater weight loss under low-carbohydrate diets would appear to be due to a lower caloric intake. Successful weight loss largely depends on restricting the intake of calories, but the supply of essential nutrients should be guaranteed. PMID- 15532737 TI - [Pituitary insufficiency following head injury--how common is it, and what is to be done?]. AB - In Germany,traumatic brain injury (TBI) has an annual incidence of approximately 200000. In contrast to earlier assumptions, pituitary insufficiency is a common complication of TBI, with a prevalence of 30-50%. Since the symptoms are often nonspecific and may be masked by the sequelae of head injury, it may go unrecognized and may possibly aggravate the symptomatology of such injury. It is therefore to be recommended that patients who suffer a head injury should be examined to exclude pituitary gland insufficiency, by measuring the basal hormone level--where necessary in combination with stimulation tests. PMID- 15532738 TI - [10 minute consultation: thrombocytosis as an incidental finding]. PMID- 15532739 TI - [Diagnosis at a glance. Nodules in the neck]. PMID- 15532740 TI - A window into the origins of the South African medical profession. PMID- 15532741 TI - HIV-infected infants born to women who tested HIV-negative during pregnancy. PMID- 15532742 TI - Dispensing--ironies and conflicts. PMID- 15532743 TI - The efficacy-effectiveness gap in PMTCT. PMID- 15532744 TI - Louis Leipoldt. PMID- 15532745 TI - Psychoneuroimmunology--mind-brain-immune interactions. PMID- 15532746 TI - What Islam does not need is a pope! PMID- 15532747 TI - Rendering unto Caesar? PMID- 15532748 TI - HIV training of traditional healers limps along. PMID- 15532749 TI - Kopanong in a spin over TOP. PMID- 15532750 TI - Time will judge 'scary' dispensing regulations. PMID- 15532751 TI - Early warning signs may signal presence of mild cognitive impairment. PMID- 15532753 TI - Marketing your medical practice--part IV. PMID- 15532752 TI - Coronary artery bypass surgery not a risk factor for dementia. PMID- 15532754 TI - Health care discrimination against the mentally ill--a comparison of private health insurance benefits for major depressive disorder and ischaemic heart disease in South Africa. PMID- 15532755 TI - A highly desired increase in 'healthy' longevity--what are the prospects? PMID- 15532756 TI - Severe high cervical spinal cord injury. PMID- 15532757 TI - What hope is there for an HIV vaccine? PMID- 15532758 TI - Asthma management in practice. PMID- 15532759 TI - A phosphodiesterase inhibitor promotes the premature development of adverse cardiac remodelling mediated by beta-adrenergic activation in hypertension. PMID- 15532761 TI - Oxytocin use in South Africa--a review. AB - OBJECTIVE: Oxytocin is one of the most frequently used drugs in labour and there are many different dosage regimens. The aim of this study was to examine the use of oxytocin by obstetricians in South Africa. METHODS: A specially designed questionnaire was drawn up and distributed to specialists according to an address list obtained from the South African Society of Obstetricians and Gynaecologists. RESULTS: Three hundred and fifty questionnaires were distributed, with 174 processed for analysis. The majority of obstetricians (70.3%) reported that they would not use oxytocin for induction of labour in a patient with a previous lower segment caesarean section, and 63.7% said that they would not consider the use of oxytocin in a patient with a multifetal pregnancy. Most respondents used oxytocin for induction of labour in multigravid patients and 91.9% also used oxytocin for augmentation in these patients. However, clinicians would not use oxytocin if the patient was a grand multipara. CONCLUSIONS: Most clinicians adhere to accepted protocols practised internationally, with a few exceptions. The use of oxytocin for both induction and augmentation of labour in women with one previous caesarean section is not practised in South Africa, despite evidence suggesting its safety. PMID- 15532760 TI - Perceptions of childhood immunisations in rural Transkei--a qualitative study. AB - OBJECTIVES: To examine perceptions of childhood illnesses, and the role of immunisation in preventing them, among caretakers of young children in Mhlakulo, a rural community in Transkei, Eastern Cape, and to suggest reasons for the low uptake of immunisations in that area. DESIGN: In-depth qualitative research using semi-structured questionnaires, focus groups, and free listing. METHODS: Detailed interviews were conducted using standardised semi-structured questionnaires. Interviews involved 60 caretakers of children aged under 5 years brought to a community health centre. Interviews were followed by two focus groups and free listing interviews to validate results of these questionnaires. RESULTS: There was widespread acceptance of the value of immunisations in preventing childhood illnesses, but only vague knowledge of why they are given, and for what illnesses. The most common knowledge was of measles and polio, but there was only limited knowledge of BCG, DPT and other immunisations. Childhood illnesses were seen as multi-causal in origin, but there was a marked absence of germ theory in explaining them. Attitudes to the use of traditional medicines in childhood were generally negative. CONCLUSIONS: Despite positive perception of immunisations, there is widespread ignorance of what they are for, and how they work. This suggests the need for increased health education, more community participation, and organisational changes in primary care clinics to make them more user friendly to caretakers of infants and young children. PMID- 15532762 TI - HIV/AIDS prevalence among South African health workers. AB - INTRODUCTION: Studies on HIV prevalence among health workers usually focus on occupational exposure to HIV. Little is known about HIV prevalence in this group. However, it is expected that HIV prevalence among health workers will reflect prevalence in their society. OBJECTIVE: To determine HIV prevalence among South African health workers. METHOD: A stratified cluster sample was drawn of 5% of health facilities in South Africa (N = 222) representative of the public and private health sectors in South Africa. The sample was designed to obtain a nationwide representative sample of medical professionals and non-professional health workers. A subsample comprising health workers in four provinces was tested for HIV status. The Orasure HIV-1 device in combination with the Vironostika HIV UNI-Form II plus O enzyme-linked immunosorbent assay (ELISA) kits were used to collect oral fluid specimens for HIV testing. RESULTS: Based on a sample of 721 health workers and a response rate of 82.5% (or 595 respondents), the study found that an estimated 15.7% (95% confidence interval (CI): 12.2 19.9%) of health workers employed in the public and private health facilities located in four South African provinces, were living with HIV/AIDS in 2002. Among younger health workers, the risk is much higher. This group (aged 18-35 years) had an estimated HIV prevalence of 20% (95% CI: 14.1-27.6%). Non-professionals had an HIV prevalence of 20.3%, while professionals had a prevalence of 13.7%. CONCLUSION: HIV prevalence among health workers in South Africa is high; this calls for the introduction of antiretroviral programmes targeting them. In addition, there is a need for the development of new policy regarding placement of infected health workers in tuberculosis (TB) wards, coupled with vigorous human resource planning to replace the health workers likely to die from AIDS. Infection control procedures also need to be reviewed. PMID- 15532763 TI - Serum procalcitonin as an early marker of neonatal sepsis. AB - BACKGROUND: It has recently been suggested that procalcitonin (PCT) is of value in the diagnosis of neonatal sepsis, with varying results. This study was to evaluate the role of PCT as a single early marker of neonatal sepsis. SETTING: Neonatal Unit, Johannesburg Hospital, and Microbiology Laboratory, National Health Laboratory Service (NHLS), South Africa. SUBJECTS AND METHODS: Neonates undergoing evaluation for sepsis between April and August 2002 were eligible for inclusion. Patients were categorised into 'no infection', 'possible infection' and 'definite infection' on the basis of C-reactive protein (CRP), white cell count (WCC), platelet count and blood culture results. PCT was correlated with infection categories. RESULTS: One hundred and eighty-three neonates were enrolled. One hundred and eighteen had no infection, 52 possible infection and 13 definite infection. PCT differed significantly among infection categories (p < 0.0001) and correlated significantly with CRP at presentation (correlation coefficient 0.404, p < 0.001) and CRP at 24 hours (correlation coefficient 0.343, p < 0.001). PCT predicted 89.5% of definite infection. Receiver operating characteristic (ROC) analysis for PCT to predict definite infection showed odds ratio (OR) 1.145 (95% confidence interval (CI): 1.05-1.25) with an area under the curve of 0.778. PCT had a negative predictive value of 0.95 (95% CI: 0.915-0.988) for definite infection. CONCLUSIONS: Although PCT was significantly related to the category of infection, it is not sufficiently reliable to be the sole marker of neonatal sepsis. PCT would be useful as part of a full sepsis evaluation, but is relatively expensive. A negative PCT on presentation may rule out sepsis, but this needs to be evaluated further. PMID- 15532765 TI - [Age-related metabolic development in rats: insulin's role]. AB - The aim of our work was to study the developmental changes in plasma and hepatic concentrations of carbohydrates, lipids, and proteins in "Wistar" rats between prepartum day 1 (fetus considered J-1) and postpartum day 133 (adult considered j+133). In addition, the evolution of insulinemia has been examined. The 21 day aged fetus (j-1), as compared to the adult (j+133) showed a lower glycemia, a higher stock of hepatic glycogen, lower rates of lipids and proteins in the plasma and in the liver. The fetal insulinemia was significantly higher than adult's. After the delivrance we have attended to an increase of the glycemia and a very marked depletion of the glycogenic stock in the liver. The plasma and hepatic lipids and proteins rose after birth. The insulinemia fell considerably and reached the lowest level. Between the 14th and 30th day of the postnatal life, a restoration of the stock of the hepatic glycogen, a normal glycemia, an elevation of the plasma and hepatic rates of proteins, and a decrease of the lipidic levels in plasma and liver were recorded. In addition the concentration of insulin increased during this period. All these values remained steady during the adult stage. We conclude that the development of rats is accompanied with several metabolic and hormonal changes. These are particularly marked at birth and weaning. PMID- 15532766 TI - [Endoscopic resection of benign prostatic hyperplasia in men over 80 years of age: a retrospective study of 184 cases]. AB - OBJECTIVE: The aim of the present study was to evaluate the outcome of transurethral resection of the prostate for benign prostatic hyperplasia in men over 80 years. MATERIAL AND METHOD: 184 men over 80 years underwent transurethral resection of the prostate for benign prostatic hyperplasia. Serious concomitant disease was found in 48.92% of cases. RESULTS: The mean operating time was 49 min ( 20 - 160mn). Others conditions found at surgery were treated during the same operation in 33.68% of cases. Peroperative complications were reported in 15 patients (8.15%) and immediate post operative complications were reported in 14 patients (7.60%), with died of 3 patients. The complications at 6 month after surgery in 172 available for followup were reported in 11 patients (6.39%). CONCLUSION: Transurethral resection of the prostate for benign prostatic hyperplasia in men over 80 years has a good outcome with an acceptable morbidity and mortality. PMID- 15532767 TI - [Predictive factors of morbidity and mortality after surgery in the elderly]. AB - The aim of the study was to determine prognostic factors of post operative morbidity and mortality for patients aged 80 years and older. We compared two groups of patients aged 80 and over operated to determine predictive factors of morbidity and mortality: group of patients who presented post operative complication within 30 days after surgery and group of patients without any complication. Comparison of the two groups for global morbidity using univariate analysis showed only one prognostic factor: surgery of diabetic foot (p = 0.034). Predictive factors of mortality according to univariate analysis were: pre operative shock (p = 0.001), abdominal wall pathology (p = 0.027), gastric or duodenal ulcer diseases (p = 0.011) and global morbidity (p = 0.006). After logistic regression, only pre-operative shock was an independent predictive factor of mortality (p = 0.0023). CONCLUSION: Risk for morbidity after surgery in the elderly 80 years and over is linked with type of surgery and not with soil. Likewise, risk for mortality is linked with advanced stage of disease that is presence of pre-operative shock and not with soil. PMID- 15532768 TI - [Clinical course of the ventilatory function in COPD: analytical study of 160 cases]. AB - The incidence of chronic obstructive pulmonary disease (COPD) is increasing particularly in developing countries. Their prognosis depend on several factors in particular the one second forced expiratory volume (FEV1) decline. To study FEV1 decline speed and factors affecting it, we carried out a retrospective study of 160 cases of COPD in stable state and heaving at least two FEV1 measures at 6 months of interval in the minimum and. The middle age was 63 +/- 12 yr, 52% was former smokers and 46% current smokers, with middle of 50 +/- 27 pack-yr. The FEV1 decline was calculated by linear regression models. The decline average in FEV1 with time was 72 +/- 55 ml/yr. Absolute value was higher than 30 ml/yr therefore more accelerated than general population in 79% of cases. We confirmed by multivariate analysis that age and smoking habit and increased the FEV1 decline. However this last correlated with FEV1% predicted. We concluded that the decline of FEV1 is higher in COPD that in the general population and slowing it needs smoking stop. PMID- 15532769 TI - [Bacteriologic features of urinary tract infections in children in the Sousse area, Tunisia]. AB - Urinary tract infection (UTI) is frequent in childhood. Our purpose is to determine the bacteriologic profile of UTI in children through a retrospective study of 1281 urinary specimens analysed in the Laboratory of Microbiology of F. Hached University hospital of Sousse between 1997 and 2002 (2000 except). The most frequent pathogens recovered were E. coli (71%), K. pneumoniae (10%) P. mirabilis (8%), Staphylococcus (1.6%), P. aeruginosa (1%) and others (2%). E. coli susceptibility to antibiotics was characterised by the high resistance percentage to amoxicillin (60%), to amoxicilline - acid clavulanic (54%) and to cotrimoxazole (40%). The resistance percentage to third generation cephalosporins, to aminoglycosides and to nitrofurane remained very low, respectively of 1.5%, 1% and 1%. High resistance rates among K. pneumoniae strains towards to amoxicillin - acid clavulanic and Cefotaxim, respectively of 63 and 39% were noticed. The resistance percentages to amikacin and cotrimoxazole were respectively of 17 and 65%, but only of 4% to nitrofurane. 70% of P. mirabilis strains were resistant to amoxicillin, 63% of them remained susceptible to amoxicillin - acid clavulanic. No resistance was shown to amikacin against 31% towards cotrimoxazole. PMID- 15532770 TI - [Management of primary gastric cancer at university hospitals in Lome. Report of 63 cases]. AB - OBJECTIVES: The objectives were to study diagnostic means, therapeutic methods and their results at the two university hospitals in LOME. METHODS: It is about descriptive retrospective study carry out at Tokoin and Campus hospitals, from 1st January 1990 to 31 December 1999 (10 years). Patients were chosen on the histological proof. RESULTS: With 63 cases colliged in 10 years on 104 cancers of digestive tract, gastric cancer occupped the uppermost (60.6%) in digestive cancers. Sex-ratio was 1.7 and average age of the patients was 52, 6 years old with extreme of 20 and 85 years old. The clinic was dominated by abdominal pains (71.4%). Vegetative ulcerous (50%) and ulcerous (21.7%) lesions were the most frequent with a predominance of pyloric antrum localisation (71.7%). Adenocarcinoma was histologic type frequently refound (87.3%). On the therapeutic way 29 patients have benefited surgical treatment, 46% of operability. Resection rate was 32% and palliative treatment was 51.7%. The postoperative mortality rate was 13.8%. CONCLUSION: Gastric cancer prognosis remain bad with a survival of 7% in one year and 21% in 3 months. PMID- 15532771 TI - [Bone mineral density in patients with Crohn's disease. Contribution of bone densitometry in 53 cases]. AB - Fifty-three patients with Crohn disease were included in a prospective study. Bone mineral density was measured at the femoral neck and lumbar spine by dual energy X-ray absorptiometry. Ten variables were analyzed in search of an association with bone demineralization; Age, sex, nutritional state, smoking, duration of the disease, hormonal status, inflammatory syndrome, site of disease: ileal, colic or ileocolic, accumulated doses of corticoids and intestinal resection. A bone demineralization was observed in 58.5% of cases. The ileum location and the corticotherapie were predictives of bone demineralization. Crohn disease is a disease at high risk of bone demineralization. Predictors of the bone loss were smoking, long duration of the disease, ileal site and systematic corticotherapy. PMID- 15532772 TI - [Laparoscopic treatment of choledocolithiasis]. AB - In a period of five years we operated 52 patients affected by choledocolithiasis. 38 patients had a laparoscopic approach, 10 patients had directly a laparotomy and 4 patients benefit of a sequential treatment. The conversion rate was 39% because we don't select candidates to laparoscopy, including even patients with acute cholangitis. Main causes of conversion were wedged calculi in the papilla, multiple stones and high inflammation of the common bile duct. Mortality and morbidity were nil. Duration of laparoscopic operation was in a mean of 165 minutes with 8.8 days of hospital stay. Two residual stones were successfully removed by endoscopic sphincterotomy. Laparoscopic treatment of choledocolithiasis is an effective, safe method; it seems to be the best therapeutic alternative. This technique needs an additional and costly instrumentation and good laparoscopic skills. In order to reduce the conversion rate it is recommended to avoid the laparoscopic approach for patients with acute cholangitis and multiple common bile duct stones. PMID- 15532773 TI - [Comparison of four techniques for the measurement of lung volumes and airway resistances by plethysmography]. AB - OBJECTIVE: To search influence of measurement technique of thoracic gas volume(Vgt) and airway resistance(Raw) by plethysmography on results. MATERIALS AND METHODS: 20 healthy subjects, aged 22.75 + 0.40 years were studied. Vgt and Raw were measured by series of gentle panting manoeuvres (GPM) or calm respiration (CR) before and after closure of a shutter valve. So, four methods were compared : method 1 (M1) CR before ahd after, method 2 (M2): CR before and GPM after, method 3 (M3) GPM before and CR after, and method 4 (M4): GPM before and after the closure of the shutter valve. Five tests were maked with each method. RESULTS: There is no significant inter-tests variability with each method. Raw(cm H2O/L/s) measured by M1 and M2 were so elevated if compared to M3 and M4 (M1 : 2.65 + 0.102; M2: 2.47 + 0.171; M3: 1.21 + 0.080; M4: 1.30 + 0.102 ; p < 0.05). There is also a significant variability intermethods in Vgt I): (M1: 3.53 + 0.188; M2 : 3.60 + 0.203; M3 : 3.91 + 0.262; M4: 3.84 + 0.247; p < 0.05). CONCLUSION: The plethysmography measurement method must be considered in the interpretation of Vgt and Raw values. PMID- 15532774 TI - [Abdomino-pelvic actinomycosis. Report of 2 cases]. AB - Actinomycosis is a chronic, granulomatous, suppurative and fistulasing infection related to a gram-positive bacteria (actinomyces israeli). Cervico-facial actinomycosis is the most common localization. The prevalence of abdomino-pelvic actinomycosis is increasing mainly with the increase of the use of intrauterin device. Its clinical presentation is variable and may mimic cancer or tuberculosis. The diagnosis of abdomino-pelvic actinomycosis is hard and most of the cases are detected during surgical exploration. We report two cases of abdomino-pelvic actinomycosis; in the first case, the disease was extended to the caecum and the abdominal wall. The skin biopsies made the diagnosis of actinomycosis, avoiding surgery. In the second case, the diagnosis of actinomycosis is made post operatively because of high suspicion of pelvic cancer. Through these two observations, we review pathogenesis of the disease, its clinical aspects and its diagnostic and therapeutic means. PMID- 15532775 TI - [Hydatic pulmonary embolism complicating a cardiac hydatic cyst. A case report]. AB - Hydatic pulmonary embolism: complication of a cardiac hydatic cyst:a case report hydatic cardiac cyst is a rare condition and represents only 0.5 to 2% of all visceral localisations of the hydatid disease. We reported a 28 year old patient with a multiple hydatic pulmonary embolism caused by the rupture of a hydatic cyst of the interventricular septum. The diagnosis was established by transthoracic echocardiography and CT Scan. Surgery was performed without delay and the outcome was good after a five month follow-up. This case illustrates the diagnostic value of the non invasive imaging means in hydatic cyst of the heart and underlines the importance of surgery realized before complications occur. PMID- 15532776 TI - [Endocarditis caused by Tropheryma whippelii]. AB - Cardiac involvement in Whipple's discase is frequent. However, its clinical occurrence without gastrointestinal symptoms is rarely described. Whipple's endocarditis occurs ollen in patients wilh native valves, without underlying, disease. It affects frequently more than one valve h, the same patient. Its clhlical presentation is generally subacute or chronic. Fever and congestive heart failure are fickle. Histological findings on the valves suggest the diagnosis, that con be confirmed by PCR technique. Treatment must be prolonged. We report a case of Whipple's endocarditis revealed by arterial embolisms and fever appeured 3 weeks after the onset. PMID- 15532777 TI - [Intracardiac thrombosis: unusual complication of Behcet's disease]. AB - Cardiac involvment is rare, and intracardiac thromboses are exceptional in Behcet's disease. We present a 29 years old male patient seen for right heart failure, thrombophlebitis and pulmonary embolism. A right atrial thrombosis was showen at the echocardiography and confirmed by computed tomography and magnetic resonance imaging. The diagnosis of Behcet's disease was established on the finding of bipolar aphtous ulcers and skin hypersensitivity. We conclude that Behcet's disease shoud be considered as a possible cause of cardiac thrombosis in the youngs men of the Mediterranean basin. PMID- 15532778 TI - [Lofgren syndrome revealed by eyelid tumor]. AB - INTRODUCTION: The Lofgren syndrom is a particular variety of sarcoidose. The ocular attack is dominated in this syndrome by anterior uveitis. More rarely the eyelids and the orbitary structures are attached. PURPOSE: In this work, we report a Lofgren syndrome case revealed by eyelid tumor. OBSERVATION: It is about 42 year-old patient who consults for a right inferior eye-lid tumefaction developped for 2 months. The diagnosis of Lofgren syndrome was evoqued because of the association of fever, arthritis, erythema nodosum, tuberculin anergia, hypercalcinuria and mediastinal adenopathy at thoracic scan-tomography. The diagnosis was then confirmed by biopsy of palpebral tumor which showed multiple epithelioid and gigantocellular granuloma without caseous necrosis. CONCLUSION: The Lofgren syndrome is a multivisceral chronic affection. The eye and its annexes constitute frequent cibles of this affection for which they react by a diverse and rich symptomalogy and which can be presented by eye-lid tumors with orbital extension. PMID- 15532779 TI - [The association of systemic lupus erythematosus and hypothyroidism. A case report]. AB - The association of systemic lupus erythematosus (SLE) and hypothyroidism is uncommon, and concerns essentially the autoimmun thyroiditis. However, the coexistence SLE-primary hypothyroidism is exceptional. We report an observation of 25-old woman, who presents a clinical form of hypothyroidism, confirmed by hormonal tests, and which is associated to polyarthritis and proximal myopathy non ameliorated by opotherapy. The explorations conclude in SLE. A favourable evolution of muscular and articular signs is obtained with corticotherapy. PMID- 15532780 TI - Prolonged survival after resection of pancreatic cancer with a solitary hepatic metastasis. PMID- 15532781 TI - Prognostic factors for hepatocellular carcinoma presenting with macroscopic portal vein tumor thrombus. AB - BACKGROUND/AIMS: This study examines the prognostic factors including radiological findings of hepatocellular carcinoma presenting with macroscopic portal vein tumor thrombus. METHODOLOGY: From September 1992 to December 2002, 107 patients with hepatocellular carcinoma and a macroscopic portal vein tumor thrombus were treated at the National Cancer Center Hospital East. Patients' characteristics and diagnostic findings of computed tomography, angiography and computed tomography angiography were analyzed to determine the factors significantly related to the patients' prognosis. RESULTS: Enhanced portal vein tumor thrombus, main tumor occupying over 40% of the liver and distribution of tumors significantly affected survival. Cavernous transformation, thread and streak signs, arterio-portal shunt, extent of tumor thrombus, grade of venous invasion and tumor size did not affect survival. Clinical findings showed that Child-Pugh classification score over 7, etiology of hepatitis, total bilirubin over 2.2 mg/dL, prothrombin time under 50% and liver transaminases over 100 IU/L were poor prognostic factors. Multivariate analysis showed that AST over 100 IU/L, viral hepatitis and tumor occupying over 40% of the liver strongly affected the prognosis. CONCLUSIONS: Based on the present results, the most strongly affected prognostic factor was liver function as indicated by high level of transaminases. Various radiological findings did not affect survival. The elevation of transaminases seemed due to destruction of hepatocytes by growing tumor and circulatory disruption due to portal vein tumor thrombus. We concluded that radiological findings of hepatocellular carcinoma presenting with portal vein tumor thrombus indicated only intrahepatic status but not survival. First treatment for hepatocellular carcinoma presenting with portal vein tumor thrombus should be to improve the liver function. Treatment against hepatitis virus might be important and patients with AST over 100 IU/L and a tumor occupying over 40% should not undergo surgical resection. PMID- 15532782 TI - Hepatic vein reconstruction using autologous vein graft for resection of advanced hepatobiliary malignancy. AB - BACKGROUND/AIMS: When advanced hepatobiliary malignancy involves the major veins that are requisite for draining venous flow from the remnant liver after hepatectomy, it may be unresectable, unless the hepatic vein involved by the tumor can be reconstructed after combined resection with hepatectomy. METHODOLOGY: Seven patients with hepatobiliary malignancy underwent hepatic vein resection and reconstruction using autologous vein grafts in our study. Five patients had patch repairs of the hepatic vein, with the gonadal vein used in 2 cases, the umbilical vein in 2, and the inferior mesenteric vein in 1. Two other patients underwent segmental reconstruction of the hepatic vein, using the inferior mesenteric vein for reconstruction of the middle hepatic vein, and the internal iliac vein for reconstruction of the right hepatic vein. Operative results and postoperative outcome, including liver function, patency, and survival, were evaluated. Literatures reporting hepatic vein reconstruction in hepatic resection, including this study, are also reviewed. RESULTS: In 6 of 7 patients, autologous vein grafts were obtained from the upper abdominal operative field without making an additional skin incision. The right internal iliac vein was used in segmental reconstruction of the right hepatic vein, and the inferior mesenteric vein was used in segmental reconstruction of the middle hepatic vein, based on caliber matching. Patch repairs of the right hepatic vein were successfully performed in 2 cases, the middle hepatic vein in 2, and the left hepatic in 1, using the umbilical vein, the gonadal vein and the inferior mesenteric vein. During hepatic vein reconstruction, total hepatic vascular exclusion was required in 2 cases, and Pringle's inflow clamp with selective clamp of reconstructed hepatic vein in 5, in order to avoid massive bleeding, congestion, and air embolism. Postoperatively, there were no remarkable complications of liver dysfunction or other organ damage encountered in any of the patients. Reconstructed hepatic veins were revealed to be patent postoperatively in all cases. Six patients survived for 60-1035 days after surgery, and 1 patient died of cancer recurrence 550 days after surgery. CONCLUSIONS: We conclude that hepatic vein reconstruction using an autologous vein graft can be safely and effectively performed with a suitable vascular control method for appropriately selected patients with advanced hepatobiliary cancer. PMID- 15532783 TI - Ambulatory esophageal pH monitoring by using a wireless system: a pilot study in Taiwan. AB - BACKGROUND/AIMS: A new wireless esophageal pH monitoring device, the Bravo system, had been tested in the US with favorable results over conventional systems. However, its clinical application in eastern countries remains limited. The aim of the study was to assess the efficacy of the Bravo system in an Asian population, and compare it with the western results. METHODOLOGY: Thirty consecutive patients with clinically evident gastroesophageal reflux disease underwent esophageal pH monitoring by using the Bravo system. The Bravo capsule capable of measuring and transmitting pH data was introduced into the squamocolumnar junction and fixed on the esophageal mucosa through a locking pin. All patients were instructed to carry the receiver for a 48-hour study. RESULTS: Twenty-nine (97%) of the 30 patients were successfully tested. In most cases (26/30, 87%) the capsules were placed without conscious sedation. All patients had successful capsule placement; however, two of them had failed the initial capsule attachment and required a second capsule. All patients had successful 24 hour recordings (30/30, 100%), while the successful 48-hour recording rate was 97% (29/30). A minor mucosal injury caused by inadvertent capsule extraction was the only complication (1/30, 3.3%). CONCLUSIONS: Compared to the western experience, the new wireless pH monitoring system achieved a comparable attaching rate and recording efficacy. Potential complication was rare and self-limited. PMID- 15532784 TI - Endoscopic ultrasonography of nonfunctioning pancreatic islet cell tumors with histologic correlation. AB - BACKGROUND/AIMS: Preoperative differentiation of benign and malignant pancreatic nonfunctioning islet cell tumors remains problematic. The present study aimed to evaluate endoscopic ultrasonography (EUS) features of benign and malignant tumors with histologic correlation. METHODOLOGY: Ten patients with surgically resected nonfunctioning pancreatic islet cell tumors were retrospectively reviewed. RESULTS: EUS demonstrated a homogenous hypoechoic (n=2) or hyperechoic (n=1) mass in the 3 benign tumors. EUS showed a hypoechoic mass with an irregular central hyperechoic portion (n=4), a hyperechoic mass with an irregular central hypoechoic portion (n=1), a cystic mass with a large irregular internal hypoechoic portion (n=1), or a hypoechoic mass within the entire lumen of the main pancreatic duct (MPD) (n=1) in the 7 malignant tumors. Histologically, the irregular central portions of the malignant tumors corresponded to necrosis, hemorrhage, fibrosis with hyalinosis, cystic degeneration, and/or calcification. Complete obstruction of the main pancreatic duct on EUS was observed in the 2 malignant tumors. The echogenicity of the tumors was closely associated with arrangement of tumor cells and quantity of fibrous stroma. CONCLUSIONS: Heterogeneous internal structures and complete obstruction of the main pancreatic duct are considered as important EUS features that are suggestive of malignancy in nonfunctioning pancreatic islet cell tumors. PMID- 15532785 TI - A comparison of serum interleukin-6 concentrations in patients treated by cholecystectomy via laparotomy or laparoscopy. AB - BACKGROUND/AIMS: Interleukin-6 (IL-6), a multifunctional cytokine, is expressed by various cells after many stimuli. This cytokine release is related, among other things, to the extent of the surgically-induced trauma. Laparoscopic cholecystectomy (LC) is a so-called "mini-invasive" surgical procedure and, on the basis of this consideration, the aim of the present prospective non randomized study, is to examine (a) whether the IL-6 is modified and how, in patients after LC compared to patients undergoing open cholecystectomy (OC), (b) whether these findings are indicative of an increased risk to develop infectious complications and whether they are therefore clinically significant. METHODOLOGY: Circulating IL-6 level was measured using a random access chemiluminescense immunoassay system in 71 patients before the operation (time 0) and 1, 2, 3, 6, 24 and 48 hours after the beginning of the operation. Thirty-five patients underwent OC and 36 LC. RESULTS: The increase in the serum IL-6 during LC was found to be significantly smaller than that during OC and resulted in a smaller extent of postoperative elevations for C-reactive protein. We recorded three cases (8.5%) of postoperative infections in the "open" group and IL-6 concentration normalized only 6 days after surgery. CONCLUSIONS: An increase in the serum IL-6 level during LC is lower in comparison to OC and results in lower postoperative elevation in C-reactive protein. Laparoscopic surgery, associated with a small skin incision and the avoidance of open laparotomy, can thus minimize surgical stress, and provide more favorable postoperative conditions for patients. Indeed excessive and prolonged post-injury elevations are associated with increased morbidity. PMID- 15532786 TI - Contribution of magnetic resonance cholangiography to the anatomic study of bile ducts. AB - BACKGROUND/AIMS: Until recently, in-depth anatomic study of the biliopancreatic ducts was only possible with injection-corrosion techniques or peroperative radiology. These were tedious and tributary of the quality of anatomic specimens. Since 1990, a new minimally invasive technique has been available: nuclear magnetic resonance cholangiography (NMRCP). The aim of the study was to evaluate the feasibility and the results of NMRCP applied to the study of the biliopancreatic tract. METHODOLOGY: NMRCP was performed in 50 healthy volunteers and results were interpreted by two readers in consensus. RESULTS: NMRCP showed the common bile, left hepatic and paramedian ducts in 100% of cases. The right hepatic duct was seen in 78% of cases. The lateral duct was seen in 98% of cases. Segmental ducts were found in 68 to 78% of cases for segments II, III, VI, VII and VIII. Only 18% of ducts for segment I were found. The gallbladder was seen in only 72% of cases. The cystic duct was clearly seen in 64% of cases. The hepato pancreatic duct was seen in 82% of cases. In 70% of cases, distribution of the hepatic ducts was modal. The remaining 30% displayed variations. CONCLUSIONS: The anatomic and radiological results reported in our study of 50 cases demonstrate the excellent feasibility and reliability of magnetic resonance cholangiography applied to the anatomic study of the bile ducts. We suggest that a wide scale multicenter study in living healthy individuals may be worth considering. PMID- 15532787 TI - Primary closure versus T-tube drainage after laparoscopic choledochotomy for common bile duct stones. AB - BACKGROUND/AIMS: To demonstrate the safety and feasibility of primary closure of the common bile duct (CBD) after laparoscopic choledochotomy in patients with CBD stones. Traditionally, the CBD is closed with T-tube drainage after choledochotomy and removal of CBD stones. However, the insertion of a T-tube is not without complication and the patients have to carry it for several weeks before removal. In the laparoscopic era, surgery is performed with minimally invasive techniques in order to reduce the trauma inflicted on patients, hasten their recovery and hence reduce the hospital stay. T-tube insertion seems to negate these benefits and we believe that primary closure can be as safe as closure with T-tube drainage. METHODOLOGY: This is a retrospective analysis of patients who underwent primary closure of the CBD after successful laparoscopic choledochotomy for ductal stones between January 2000 and December 2003. A concurrent control group of patients who underwent T-tube drainage was used for comparison. RESULTS: Of the 64 patients that underwent laparoscopic exploration of the CBD, 24 (37%) underwent transcystic duct approach and 40 (63%) underwent choledochotomy. There were three open conversions (5%). Stone clearance was achieved in all patients with successful laparoscopic choledochotomy (100%). Of the 38 successful laparoscopic choledochotomies, 12 had primary closure of the CBD and 26 had closure with T-tube drainage. There was no mortality in both groups. One patient in the primary closure group suffered from paralytic ileus and small subhepatic collection which was treated conservatively. The median operative time (90 vs. 120 minutes, p=0.002) and postoperative stay (5 vs. 8.5 days, p=0.003) were shorter in the primary closure group when compared with the T tube group. CONCLUSIONS: Primary closure of the CBD is feasible and as safe as T tube insertion after laparoscopic choledochotomy for stone disease. PMID- 15532788 TI - Factors influencing recurrence after surgical treatment for T2 gallbladder carcinoma. AB - BACKGROUND/AIMS: The overall outcome of T2 gallbladder carcinoma has not been favorable, although there is a modest hope for long-term survival after radical resection. The aim of this study was to examine factors influencing postoperative disease-free survival of patients with T2 gallbladder carcinoma to clarify optimal treatment. METHODOLOGY: Of 53 patients with gallbladder carcinoma who had undergone surgical resection from 1985 to 2000, 22 had T2 carcinoma histologically proved. The significance of variables for disease-free survival was examined retrospectively by the Kaplan-Meier method and the log-rank test. RESULTS: There were 16 patients with stage II (T2N0M0), 6 with stage III (T2N1M0) disease. Eleven patients were treated by extended cholecystectomy with resection of the extrahepatic bile duct, 10 patients underwent extended cholecystectomy without resection of the extrahepatic bile duct, and 1 patient underwent cholecystectomy. All patients underwent lymph node dissection in the hepatoduodenal ligament, below the pancreatic head, and along the common hepatic artery. Lymph node metastasis was present in 6 patients. Lymphatic, venous, and perineural invasions were found in 9, 4, and 4 patients, respectively. The absence of lymphatic invasion was a significant factor related to good postoperative disease-free survival (5-year disease-free survival rate, 88.9% vs. 31.3% in the presence of lymphatic invasion). Lymph node, venous, or perineural invasion, and surgical procedure were not significant factors to good postoperative disease-free survival. CONCLUSIONS: For patients with T2 gallbladder carcinoma, the presence of lymphatic invasion is an unfavorable prognostic indicator that calls for additional treatment after radical surgery. PMID- 15532789 TI - Patterns and relevant factors of tumor recurrence for extrahepatic bile duct carcinoma after radical resection. AB - BACKGROUND/AIMS: The complete surgical removal of bile duct cancer offers the only chance of cure. Long-term survival, however, is very limited because of frequent recurrence after surgery. As it is important to know the mode of recurrence and its relevant factors for early detection, prevention, and selecting an ideal strategy for adjuvant therapy, we determined the frequent mode of recurrence after curative surgery and analyzed its risk factors. METHODOLOGY: Eighty-three patients who underwent radical resection for bile duct cancer were evaluated. The tumors comprised 24 proximal and 59 middle or distal bile duct carcinomas. The clinicopathologic findings and the modes of recurrence were reviewed. RESULTS: Tumor recurrence developed in 18 of 24 proximal and 34 of 59 non-proximal cancers at an average interval of 16.7 and 17.8 postoperative months, respectively. T-stage (p=0.031) and age (p=0.019) were independent factors affecting recurrence on multivariate analysis. The factors, however, were different according to tumor location that while histological grade was the only factor for proximal tumor, nodal status and age were independent factors for non proximal tumors. The precise recurrent sites were identifiable in 14 proximal and 24 non-proximal lesions. While the predominant pattern of recurrence was locoregional in most proximal tumors (13/14), non-proximal tumors showed frequent recurrences at distant sites including liver, peritoneum, and lung (13/24). The median survival was longer in non-proximal (45.3 months) than proximal tumor (18.0 months), in T0,1-tumor (>120 months) than T2,3-tumor (20.4 months), and in N0-tumor (60.2 months) than N1,2-tumor (13.0 months). The 5-year survival rates were also higher in non-proximal cancers (46.7% vs. 32.0%). CONCLUSIONS: Our results suggested that the need for postoperative adjuvant therapy, and strategies to detect and to prevent recurrence should be individualized not only by stage but also by tumor location to improve survival in patients with extrahepatic bile duct cancer. PMID- 15532790 TI - Analysis of the motor function of the human sphincter of Oddi by endoscopic retrograde cinecholangiography gated by manometry--a report of a case. AB - Although the motor function of the sphincter of Oddi (SO) has been clearly identified by endoscopic SO manometry (ESOM), the physiologic role of the phasic contractions of the SO remains unsettled in humans. The aim of this study was to correlate SO motor activity measured by ESOM with bile flow characteristics determined by simultaneously recorded endoscopic retrograde cinecholangiography. We investigated a 55-year-old female patient by means of ESOM. During the station pull-through recording, the ESOM catheter was withdrawn into the SO zone and retained there for 15 min. The pressures transmitted by the external transducers and the enlarged video picture of the choledochoduodenal junction from the X-ray fluoroscopic monitor (25 digital pictures/sec) were recorded simultaneously on the computer system with a time-correlated basis. During the analysis without taking note of the cinefluoroscopic events, we selected different manometric periods manually, such as the pressure wave of the SO phasic contraction, no SO phasic activity and the first second of the beginning of the next phasic contraction. Cumulative cinecholangiographic pictures were then constructed by the computer for each period, at a frequency of one frame/sec to create representative sum-of-pictures for each manometric period. By means of the application of manometrically gated cinecholangiography, we succeeded in demonstrating an exact time correlation between the SO systolic and diastolic movements on cinecholangiography and the pressure recording detected by ESOM in humans. PMID- 15532791 TI - Partial separating gastrojejunostomy for incurable cancer of the stomach or pancreas. AB - BACKGROUND/AIMS: Advanced stomach or pancreas cancer with antral obstruction has been treated by gastrojejunostomy. The delayed return of gastric emptying, however, frequently occurs. The Devine exclusion procedure has been reported to be the better bypass operation in terms of oral intake, but it needs a drainage tube. In cases where the lesser curvature is invaded, this operation should be avoided. A method of gastroenterostomy, which is safe and shows good outcomes concerning oral intake, is desired. METHODOLOGY: Among 15 patients with advanced stomach or pancreas cancer, 8 received conventional gastrojejunostomy (CG Group), 3 Devine exclusive gastrectomy with a drainage tube (DE Group) and 4 partial separating gastrojejunostomy (PG Group). The partial separating gastrojejunostomy was performed as follows. The stomach was partially partitioned using GIA from the side of the greater curvature. The posterior side of the proximal stomach was anastomosed with the proximal jejunum using a circular stapler instrument. RESULTS: All patients in the DE and SG Groups could eat regular or semi-regular meals. The bleeding from tumor in the DE Group was less than that in the SG and CG Groups. CONCLUSIONS: In cases where the lesser curvature is invaded by tumor or lymph node metastasis, partial separating gastrojejunostomy would be recommended as a substitute for the Devine procedure. PMID- 15532792 TI - Overexpression of cyclooxygenase-2 and tumor angiogenesis in human gastric cancer. AB - BACKGROUND/AIMS: Cyclooxygenase-2 (COX-2) protein is overexpressed in various cancers, including esophageal, gastric, colon, and pancreatic. To better comprehend the role of COX-2 in gastric cancer, especially with regard to angiogenesis, we investigated COX-2 and vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) in 108 patients with gastric cancer. METHODOLOGY: We used immunohistochemical analysis of formalin-fixed tissues of gastric cancer. RESULTS: Expression of COX-2 showed diffuse staining in the cytoplasm of tumor cells, however, no staining in normal epithelial cells. Of the 108 tumors examined, 71 (65%) were positive for COX-2 expression, the VEGF positive cases numbered 43 of 108 cases (39.8%). The intensity of COX-2 expression did not correlate with any clinicopathological characteristics. The positive rate of VEGF expression in COX-2-positive cases was significantly higher than in COX-2-negative ones (47.9% vs. 24.3%, P<0.05). MVD in COX-2-positive cases was significantly higher than in COX-2-negative ones (22.0+/-7.8 vs. 18.5+/ 7.5/1 mm2; P<0.05). CONCLUSIONS: Our study provides evidence that COX-2 is closely related with angiogenesis. PMID- 15532793 TI - Transthoracal partial resection or local excision of small esophageal carcinoma with a radical mediastinal lymphadenectomy--proposal of limited operation for postoperative quality of life. AB - BACKGROUND/AIMS: Controversy remains regarding which treatment should be employed for superficial esophageal cancer. The treatment modalities for such superficial cancers vary from a local excision, including an endoscopic mucosal resection (EMR) to an extended radical esophagectomy (three-field lymphadenectomy). In the current report, we proposed the use of either a transthoracal partial resection or local excision of the esophagus with a radical mediastinal lymphadenectomy for the patients with small superficial esophageal squamous cell carcinoma, in order to perform effective radical cancer surgery and maintain the quality of life (QOL) of the patients. METHODOLOGY: Surgical procedures: After thoracotomy and detaching of the esophagus from the surrounding tissues, the proximal and distal resection lines were determined by careful palpation of the clips within the esophagus which had been placed during the preoperative endoscopic examination. Thereafter, a mediastinal lymph node dissection, including the bilateral recurrent nerve nodes and paratracheal nodes, was performed. After this procedure, intrathoracic esophageal end-to-end anastomosis was performed in the ordinary manner. We performed this procedure on three patients with small localized esophageal cancer. RESULTS: Operations were safely performed on these three patients and postoperative quality of life was almost satisfactory. CONCLUSIONS: Transthoracal partial resection or local excision with a radical mediastinal lymphadenectomy was useful for the patients with small localized esophageal carcinoma. PMID- 15532794 TI - Differences in clinicopathological findings, cell kinetics and p53 expression between early gastric cancers with and without Helicobacter pylori infection. AB - BACKGROUND/AIMS: Helicobacter pylori (Hp) infection has been suggested to be a risk factor for gastric carcinogenesis. The aims of this study were to clarify differences in clinicopathological features, tumor cell kinetics and p53 expression between early gastric cancers developing within and at a distance from Hp actively infected mucosa and those away from the infected area. METHODOLOGY: A total of 91 consecutive patients who were diagnosed as having early gastric carcinoma were enrolled in this study. Phenol red solution and urea were sprayed over the surgically resected stomachs (the PR test). Tumors included in and away from positive (red color) areas on PR testing were considered as infected and non infected cases, respectively, and compared for Ki67 (proliferative activity), ssDNA (apoptotic activity) and p53 immunoreactivity. RESULTS: The average age of the infected cases (n=46) was 9 years younger than that for their non-infected counterparts (n=45; P<0.0001). Depressed macroscopic and diffuse histologic types were more prevalent in the infected cases (P<0.05 and P<0.001, respectively). In neither intestinal nor diffuse histologic types were any significant differences in Ki67, ssDNA or p53 immunoreactivity apparent between the infected and non infected cases. CONCLUSIONS: Cases of early gastric cancer developing within and at a distance from Hp infected mucosa are clinicopathologically different although the presence of bacteria in the surrounding mucosa does not appear to affect tumor cell kinetics or p53 expression. PMID- 15532795 TI - Simultaneous transfer of vascular endothelial growth factor and hepatocyte growth factor genes effectively promotes liver regeneration after hepatectomy in cirrhotic rats. AB - BACKGROUND/AIMS: Liver regeneration in a cirrhotic liver is unsatisfactory. In the course of liver regeneration, non-parenchymal cells such as sinusoidal endothelial cells as well as hepatocytes increase in number while the liver structure and physiological functions are maintained. The aim of this study was to examine whether sufficient liver regeneration could be obtained by the simultaneous, preoperative injection of recombinant adenoviral vectors encoding human vascular endothelial growth factor (VEGF), a potent mitogen for sinusoidal endothelial cells, (pAxCAVEGF) and rat hepatocyte growth factor (HGF), a potent mitogen for hepatocytes, (pAxCAHGF) in 70% hepatectomized cirrhotic rats. METHODOLOGY: Forty-eight hours before 70% hepatectomy, dimethylnitrosamine induced cirrhotic rats were infused intravenously with pAxCAVEGF or with pAxCAVEGF and pAxCAHGF, or with a control virus encoding Escherichia coli beta galactosidase (pAxCALacZ). RESULTS: Strong VEGF mRNA expressions were shown in the livers of VEGF and VEGF/HGF-treated animals. The plasma HGF concentrations in the VEGF/HGF-treated rats were elevated compared with the other groups. Proliferating cell nuclear antigen immunostaining showed increased labeling indices of hepatocytes in the VEGF/HGF-treated rats at 24 and 48 h after hepatectomy. PCNA labeling indices of SECs were increased in the VEGF and VEGF/HGF-treated rats compared with the control animals at 24 and 48 h after hepatectomy. Moreover, the hepatic regeneration rate after hepatectomy was significantly augmented by the VEGF and VEGF/HGF treatment. CONCLUSIONS: Simultaneous preoperative injection of recombinant adenoviral vectors encoding VEGF and HGF effectively stimulates liver regeneration in cirrhotic rats. PMID- 15532796 TI - What types of colorectal cancer overexpress the MAGE protein? AB - BACKGROUND/AIMS: Tumor-specific antigens such as Melanoma-associated antigens could be attractive targets for immunotherapy. It will be a great help for cancer immunotherapy to distinguish what types of tumor expresses tumor-specific antigen. METHODOLOGY: We investigated the expression pattern of MAGE-A1 by immunohistochemical typing methods in human colorectal cancers obtained from consecutive patients undergoing surgical treatment at the hospital of the University of Tokyo. RESULTS: In 35 of 89 cases (39%), MAGE-A1 positive immunoreactivity was detected in the malignant glands. MAGE-A1 positive immunoreactivity was significantly higher among the patients under the age of 70 than among those 70 years of age and older (p=0.04). Among the patients under the age of 70, the tumors located in the distal colon (descending colon, sigmoid colon and rectum) showed significantly more positive MAGE-A1 immunoreactivity than those in the proximal colon (cecum, ascending colon and transverse colon) (p=0.04). The expression pattern of MAGE-A1 was not associated with gender, size, depth, histological type, vessel invasion, lymphatic invasion, lymph nodal invasion or stage of disease. CONCLUSIONS: Relatively better results can be expected with MAGE-A1 immunotherapy among patients with distal colon cancer under the age of 70. PMID- 15532797 TI - Nuclear survivin expression predicts poor outcome in cholangiocarcinoma. AB - BACKGROUND/AIMS: Survivin is an anti-apoptotic protein expressed in cancer and may have prognostic value. Our study examined the prognostic role of survivin in biliary cancer. METHODOLOGY: Twenty-four consecutive cases of cholangiocarcinoma were studied. Immunohistochemistry was performed using a monoclonal antibody to survivin. Survivin expression was described as absent or weak and strong. RESULTS: Median age was 68 years (range 40 to 77). There were 16 females and 8 males. SEER (Surveillance, Epidemiology and End Results) staging was local in 2, regional in 15 and distant in 7. Treatments included chemotherapy (n=3), surgery (n=9), combined modality (n=10) or no therapy (n=2). Cytoplasmic and nuclear survivin expression was seen in 13 and 11 patients respectively. Strong cytoplasmic survivin expression was seen in 6 cases and strong nuclear survivin in 4. Patients with strong nuclear survivin had a median survival of 11 months, significantly lower than for patients with weak nuclear survivin expression (20 months, p=0.033). Multivariate analysis using the Cox-proportional hazards model identified 4 important prognostic predictors: nuclear survivin (P=0.022), presence of metastasis (P=0.025), age (P=0.019) and use of combined modality therapy (P=0.006). CONCLUSIONS: Nuclear survivin expression in cholangiocarcinoma may identify those with a poor prognosis. PMID- 15532798 TI - Argon plasma coagulation for the treatment of early gastric cancer. AB - BACKGROUND/AIMS: Endoscopic mucosal resection (EMR) is widely used to treat early gastric cancer and is considered safe and effective. However, its indication range is limited. Other endoscopic treatment options are needed for patients with surgical risks. The aim of the study was to evaluate the safety and effectiveness of argon plasma coagulation (APC) for the treatment of early gastric cancer. METHODOLOGY: APC was done in 23 patients (mean age, 77.5 years) with early gastric cancer. The depth of tumor invasion, estimated primarily by endoscopic ultrasonography, was mucosal in 19 patients and submucosal in 4. EMR was not indicated in 4 cases, and 14 cases were not successfully treated by EMR alone. All patients were followed up for more than 12 months (median, 42.0+/-20.8 months). RESULTS: Fifteen patients had no recurrence and survived. Four patients had no recurrence, but died of causes other than gastric cancer. Cancer recurred in four patients. Recurrence was managed by repeated treatment with APC, with no technical problems. No serious complications occurred. CONCLUSIONS: APC is useful for follow-up treatment of early gastric cancer after EMR. APC may also be effective for radical treatment of early gastric cancer, especially in elderly patients and patients in whom surgical intervention is contraindicated. PMID- 15532799 TI - Experimental study of a novel thermotherapy for hepatocellular carcinoma using a magnesium ferrite complex powder that produces heat under a magnetic field. AB - BACKGROUND/AIMS: The anti-cancerous effect on hepatocellular carcinoma of a newly established form of thermotherapy, which uses an implant heating system, was evaluated. As a new material for application in hyperthermia, the authors developed a powder type Mg-ferrite complex that produces heat under a relatively low-power magnetic field. METHODOLOGY: This material suspended in Lipiodol was injected into tumors on the backs of mice that consisted of human hepatocellular carcinoma cells. Hyperthermia was performed by directing a magnetic charge on tumor-bearing mice that contained the Mg-ferrite complex. The temperature of the tumor was kept at 42-43 degrees C, while the magnetic field power ranged from 50 to 80G. RESULTS: A 10-min hyperthermia treatment was insufficiently effective against tumor growth. Systemic injection of doxorubicin (ADM) before hyperthermia appeared to enhance the anti-cancerous effect, but the difference was little and did not reach a statistically significant level (repeated measure analysis of variance). The anticancerous effect of hyperthermia for 15 minutes, in contrast, was marked. The nodules had almost completely disappeared by the end of the experiment. CONCLUSIONS: In conclusion, it is suggested that hyperthermotherapy using this newly developed Mg-ferrite complex might become an option for low invasive therapy for advanced hepatocellular carcinoma in humans. PMID- 15532800 TI - Interleukin-6 augments hepatocyte growth factor-induced liver regeneration; involvement of STAT3 activation. AB - BACKGROUND/AIMS: In liver regeneration, quiescent hepatocytes need to be primed before they can fully respond to growth factors such as hepatocyte growth factor (HGF). Recently, interleukin-6 (IL-6) has been shown to play an important role in initiating liver regeneration via activating signal transducer and activator of transcription 3 (STAT3). We therefore investigated the effect of IL-6 on liver regeneration and examined STAT3 activation following IL-6 and/or HGF treatment in rats. METHODOLOGY: The animals underwent portal branch ligation (PBL) of the left lateral and median branches. They were intravenously treated with either IL-6 (500 microg/kg), HGF (50 microg/kg), both, or vehicle alone on and every 12 h after PBL. The degree of compensatory hypertrophy in unoccluded lobes was examined by measuring the wet weight ratios of the unoccluded lobes to the whole liver and the 5-bromo-2'-deoxyuridine labeling index of hepatocytes in each group. STAT3 expression in regenerating liver was examined by Western blotting and immunohistochemistry. RESULTS: Coadministration of IL-6 and HGF most effectively increased both the wet weight of the unoccluded lobes and the hepatocellular DNA synthesis. HGF as well as IL-6-treated animals showed moderate STAT3 activation in hepatocytes throughout the experiment. CONCLUSIONS: IL-6 augmented HGF-induced liver regeneration and hepatocellular replication. Liver regeneration stimulated by HGF treatment may be closely involved with STAT3 activation in a different manner induced by IL-6 treatment. PMID- 15532801 TI - A new test to avoid arterial complications during pancreaticoduodenectomy. AB - Celiac trunk or superior mesenteric artery stenosis are usually innocuous conditions. In such patients, arterial blood supply to the stomach, spleen, liver and bowel is sustained through extraordinarily well-developed pathways through the pancreas. If division of these collateral vessels is necessary during a surgical procedure such as pancreaticoduodenectomy, life-threatening celiac organ or bowel ischemia may occur. The authors describe a new test, using pancreatic inflow occlusion, to reliably identify celiac trunk or superior mesentery artery stenosis. The authors describe two cases of celiac axis occlusion and one case of superior mesenteric artery stenosis. In all three presented cases the gastroduodenal artery clamping test was negative and ischemia was only noticed after pancreatic section, suggesting that in severe occlusions this test may fail in diagnosing the vascular abnormality. All patients were successfully treated by revascularization with no operative mortality. If the diagnosis is unsuspected and in cases where appropriate angiographic studies have not been obtained before pancreatic resection, a test occlusion of the gastroduodenal artery should always precede its ligation. However, this test may not be effective in all cases and in instances where high suspicion of celiac axis or mesenteric stenosis is present, other maneuvers, such as pancreatic inflow test, could be helpful for the diagnosis of these rare and morbid situations. PMID- 15532802 TI - Successful treatment for combined pancreatoduodenal injury by a second-stage pancreatojejunostomy following pancreatoduodenectomy. AB - A case of combined pancreatoduodenal injury requiring pancreatoduodenectomy is described. A two-stage approach was used, which included a pancreatoduodenectomy followed by a tube pancreatostomy for complete external drainage of pancreatic juice and subsequent second-stage pancreatojejunostomy. This approach, first advocated by Miyagawa et al., enabled us to handle the normal soft pancreas without any complications. It is suggested that the described second-stage pancreatojejunostomy can be considered as an alternative method of restoring pancreatoenteric continuity following pancreatoduodenectomy in emergency cases. PMID- 15532803 TI - Markedly elevated alpha-fetoprotein levels without hepatocellular carcinoma. AB - Although low-grade elevations of AFP are associated with benign liver disease including acute and chronic hepatitis and cirrhosis, AFP values above 400 ng/mL are often used as a surrogate marker for HCC. The case of a 45-year-old Caucasian woman, who while receiving interferon therapy for HCV, was found to have a marked elevation of her serum AFP level, is reported. Her baseline AFP of 7.0 ng/mL increased progressively over three months to a peak value of 734.5 ng/mL. Initial imaging was normal. Three months later, a CT scan detected two focal lesions in the dome of the liver. A post-ethiodol hepatic CT scan revealed persistent uptake at this site but fluoroscopy-guided liver biopsy failed to identify a HCC. Normalization of her AFP level occurred while she continued to receive IFN for HCV. The patient has remained stable over more than three years of follow-up. While others have reported AFP elevations in cirrhotics with HCV without evidence of HCC, to our knowledge there are no reports of an individual receiving IFN treatment for hepatitis C with AFP levels that rose over several months in the absence of a HCC. PMID- 15532804 TI - Postoperative chemoradiotherapy in rectal cancer. Late results of a pilot study. AB - BACKGROUND/AIMS: It has recently been proven that postoperative radiotherapy combined with fluorouracil showed an increase of survival and local control in patients with rectal cancer. However, hematological and intestinal toxicity also increased. Experimental and clinical studies showed an increased radiation effect with an acceptable toxicity by delivering drug via a continuous intravenous infusion. METHODOLOGY: From 1988 to 1998, 80 patients radically operated on for stages B2-C rectal cancer were irradiated with 3 fractions of 100 cGy per day to a total dose of 5,600 cGy. 34 out of these 80 patients underwent postoperative radiotherapy alone and 46 received radiotherapy combined with concomitant protracted infusion of fluorouracil at doses of 250 mg/m2 per day. RESULTS: After a median follow-up of 54 months, the 5-year overall and disease-free survival were 59% and 54%, respectively, in the combined modality group, as compared to 42% and 34%, respectively, in the radiation alone group. The differences were not significant, but the incidence of local relapse and patients' survival showed a better trend for combined approach. CONCLUSIONS: The data from international literature are in favor of a combined approach, both in preoperative and postoperative treatment of advanced rectal cancer. Adjuvant therapy must be re evaluated in trials using total mesorectal excision as the standard operative technique. PMID- 15532805 TI - Clinical significance of virtual colonoscopy (CT colonography) with special reference to polyp morphology. AB - BACKGROUND/AIMS: Computed tomographic colonography [virtual colonoscopy (VC)] is a new imaging method that may contribute to the detection of colorectal neoplasms. The aim of this study was to investigate the clinical significance of VC for the diagnosis of colorectal tumors with special reference to their morphology. METHODOLOGY: Sixteen patients with colorectal tumors were enrolled in this study. Colonic preparation was performed using magnesium sulfate, and scopolamine butylbromide was injected before air insufflation from the anus. VC was performed with an Aquilion multislice CT system (Toshiba) and Workstation M900 maximum (ZIO). The results were compared with those of conventional colonoscopy (CC). In six cases, total colonoscopy was not possible due to tumor stricture. RESULTS: The number of lesions detected by CC was 58; 48 early carcinomas/adenomas (group A) and 10 advanced carcinomas (group B). All group B lesions and 43.8% (21 lesions) of group A lesions were identified by VC. The specificity of VC for group A and B lesions was 45.8% and 100%, respectively. There was no significant difference in detection rate for group A lesions by subclassification according to their morphology: pedunculated; 22% (2/9), semipedunculated; 50% (6/12), sessile; 29% (4/14), superficially elevated; 56% (5/9), and superficially elevated morphology with central depression; 100% (4/4). The mean diameter of the lesions that were detected and not detected by VC was 7+/-8.0 mm (range: 3-30 mm) and 8+/-5.8 mm (range: 2-20 mm), respectively (P=0.90). CONCLUSIONS: Polyps with superficial morphology could be detected by VC as well as those with protuberant morphology. According to evidence that neither the morphology nor the size of a lesion was a significant factor for detection by VC, the authors consider that the diagnostic ability of VC may be improved by better preparation. PMID- 15532806 TI - Prognostic significance of total lymph node number in patients with T1-4N0M0 colorectal cancer. AB - BACKGROUND/AIMS: The presence of lymph node metastases is the main prognostic factor in patients with colorectal cancer. Controversy exits relating to the minimum number of lymph nodes to be harvested for accurate staging in patients classified as stage I and II. The purpose of this study was to define a cut-off value for the number of lymph nodes examined and determine if this minimum number could have any impact on prognosis of node negative patients. METHODOLOGY: A total of 301 T1-4N0M0 colorectal cancer patients were reviewed. The clinical and pathological factors considered in statistical analysis were age, gender, tumor location, depth of invasion, differentiation and the number of examined lymph nodes. RESULTS: The median number of harvested lymph nodes was 10 and the 5-year overall survival rate was 62%. Univariate analysis revealed that patients with well differentiated, right-sided, T1-2 tumors and those with more than 11 nodes examined had improved survival compared to their counterparts. In Cox regression analysis, the same variables except tumor location independently affected the prognosis, where the number of lymph nodes recovered showed the highest significance (p=0.0008; RR: 2.8; 95% CI: 1.6-5.2). CONCLUSIONS: Patients classified as T1-4N0M0 colorectal carcinoma with more than 11 lymph nodes examined may be considered as more accurately staged and have better prognosis than those with lower number of lymph nodes recovered. PMID- 15532807 TI - The value of omentoplasty in protecting colorectal anastomosis from leakage. A prospective randomized study in 126 patients. AB - BACKGROUND/AIMS: Anastomotic leakage is a major cause of mortality in colorectal surgery. Several methods have been evaluated in order to prevent anastomotic leakage. To decrease the rate and severity of anastomotic leakage, omentoplasty (OP) has been proposed by several authors on the basis of experimental and clinical studies. A prospective, randomized trial was designed to study the influence of omentoplasty on anastomotic leakage after colorectal resection. METHODOLOGY: One hundred and twenty-six patients undergoing elective or emergency surgery for malignancy, benign tumor, diverticular disease and other were randomly assigned to omentoplasty (OP group) or not (NO group). The primary end point was the rate of clinical and radiological anastomotic leakage. Both groups were comparable in terms of demographic data, preoperative characteristics and intraoperative findings. RESULTS: Eighteen patients (14.3%) had anastomotic leakage, 4 (6.4%) in the OP group and 14 (21.9%) in the NO group. Significant differences (P<0.05) between the two groups were also found in terms of repeat operation (3.2% vs. 14.1%) and deaths (3.2 vs. 7.8%). Other factors associated with anastomotic leakage were the distal site of anastomosis (<5 cm from anal verge) and the emergency. CONCLUSIONS: Omental wrap, with its mechanical and biological properties, seems to be effective in lowering the rate and the severity of anastomotic leakage after colorectal surgery. PMID- 15532808 TI - Clinicopathological characteristics of poorly differentiated adenocarcinoma of the colon and rectum. AB - BACKGROUND/AIMS: The clinicopathological significance of poorly differentiated adenocarcinoma (Por) of the colorectum remains controversial. The purpose of this study was to investigate the clinicopathological characteristics of Por by comparison with well (Well) and moderately differentiated adenocarcinoma (Mod) of the colorectum and by subclassification of Por into three types according to the amount of fibrous stroma in tumor tissue. METHODOLOGY: Eleven patients with Por, 64 with Well and 254 with Mod were clinicopathologically compared. RESULTS: Por was located predominantly in the right side of the colon. The maximal size of the tumors in Por (72.3+/-25.0 mm) was significantly larger than in Well (42.2+/-26.8 mm) and Mod (52.2+/-22.8 mm) (p=0.0009, 0.0047). The frequency of lymph node metastasis in Por was significantly higher than in Well (p=0.0009). The five-year survival rate for patients with Por was 45.5%, for Well was 71.4% and for Mod was 59.5% (NS). The medullary type of Por had a good prognosis. CONCLUSIONS: Por proliferated and metastasized more rapidly than Well. The subclassification of Por according to the amount of fibrous stroma in tumor tissue could play an important role in the clinicopathological study of colorectal carcinoma. PMID- 15532810 TI - Treatment of irritable bowel syndrome with colonic pacing: evaluation of pacing parameters required for correction of the "tachyarrhythmia" of the IBS. AB - BACKGROUND/AIMS: A recent study of the electromyographic (EMG) activity of irritable bowel syndrome (IBS) has shown that the frequency, amplitude and conduction velocity of the slow waves (SWs) of the sigmoid colon (SC) were significantly higher in IBS patients than in the healthy volunteers. The SW rhythm was irregular. A "tachyarrhythmic pattern" was characteristic of the IBS. The SC pressure in the IBS was also significantly higher than that of the healthy controls. We suggested that the cause of IBS is related to an aberrant focus in one or more of the colonic pacemakers which possibly triggers abnormal impulses to the colon. We hypothesized that stimulation of the pacemaker which delivers electric waves to the SC, may correct the abnormal electric waves and eliminate the IBS symptoms. In this communication we tried to define the adequate pacing parameters necessary for normalization of the tachyarrhythmic pattern of the electric waves in IBS. METHODOLOGY: Nineteen subjects with IBS were divided into a study group (age 48.6+/-9.8 years; 7 women, 4 men) and a control group (age 47.6+/-9.2 years; 5 women, 3 men). The study also included 8 healthy volunteers (47.9+/-9.7 years; 5 women). Three 28-gauge cardiac pacing electrodes were used: one for pacing applied to the pacemaker at the colosigmoid junction (CSJ) and 2 for recording applied to the SC mucosa. In the study group, the CSJ electrode was stimulated using an electrical stimulator which delivered a constant current. The optimal pacing parameters had been determined after repeated trials with different variables. In the control group, recording was done without pacemaker activation. The SC pressure was measured by a 10-F saline-perfused tube. RESULTS: In the healthy volunteers, the basal SWs were regular and followed or superimposed by action potentials (APs). Pacing produced a significant increase in the SW variables and SC pressure; the latency was 20.3+/-3.6 s. The study and the control group exhibited a basal tachyarrhythmic pattern and a significantly higher SC pressure than the healthy volunteers. Pacing of the study group effected lowering of the SW variables and SC pressure which did not show a significant difference against those of the healthy volunteers at rest. The optimal pacing parameters comprised an amplitude of 6 mA, a pulse width of 150 ms and a 25% higher frequency than that of the already recorded basal colonic waves. The control group showed no change in the tachyarrhythmic pattern. CONCLUSIONS: CS pacing parameters were identified and succeeded in normalizing the tachyarrhythmic pattern of the IBS. We suggest that this method be used for the treatment of patients with IBS when other measures have failed to cure the condition. PMID- 15532809 TI - Preoperative chemoradiation therapy for low rectal cancer. Impact on downstaging and sphincter-saving operations. AB - BACKGROUND/AIMS: Multimodality treatment is the preferred strategy in the management of low rectal adenocarcinoma. Preoperative chemoradiation therapy (CRT) associated with radical surgery may result in significant tumor downstaging, local control of the disease and high rates of sphincter-saving operations. METHODOLOGY: One hundred consecutive patients with advanced and resectable distal rectal cancer treated either by surgery alone or by preoperative chemoradiation followed by surgery were reviewed. Both groups were similar in tumor and patients characteristics. Treatment strategy was randomly selected. RESULTS: Mean tumor size in Group 1 (surgery alone) was 6.1 cm and 3.3 cm in Group 2 (CRT). In Group 1 (surgery alone), nine patients had stage I (18%), 20 had stage II (40%) and 21 had stage III disease (42%). In Group 2 (CRT), 6 patients had stage 0 (12%), 21 had stage I (42%), 12 had stage II (24%) and 11 had stage III disease (22%). Furthermore, for each T (except for T0-1) the presence of lymph node (LN) metastasis was significantly more frequent in Group 1. In Group 2 (CRT), there was a mean reduction of 27% of the initial tumor size. This reduction was observed in 58% of patients in this group (CRT). Finally, sphincter-saving operations were more frequently performed in Group 2 (CRT) even though tumor differentiation and distance from the anal verge were similar in both groups. CONCLUSIONS: Preoperative CRT followed by radical surgery is an effective treatment strategy for distal rectal cancer resulting in significant tumor size reduction and downstaging. Consequently, these effects may be responsible for higher rates of sphincter-saving operations performed in this subset of patients. PMID- 15532812 TI - Phonatory function following esophagectomy for esophageal cancer. AB - BACKGROUND/AIMS: Quality of life can be adversely affected in many patients who suffer phonation disorders such as hoarseness and dysphonia following esophagectomy. The present study investigated postoperative phonation disorders in 15 patients who underwent esophagectomy for esophageal cancer. METHODOLOGY: None of the patients had signs of hoarseness before or after surgery. Aerodynamic testing to assess phonatory function testing and laryngoscopy for observing laryngeal movements were performed before and after surgery. As a control, the same tests were conducted in 20 patients treated for gastric cancer by gastrectomy. RESULTS: For esophagectomy patients, mean postoperative flow rate was significantly increased and maximum postoperative phonation time was significantly decreased after operation. Laryngoscopy confirmed postoperative paralysis of left laryngeal movements and excessive adduction of the right, unaffected vocal cord during phonation in 8 of 15 esophagectomy patients, although hoarseness was not reported by any patient. No significant changes were observed for mean postoperative flow rate or maximum postoperative phonation time following surgery in gastrectomy patients. CONCLUSIONS: Surgical procedures in the vicinity of the recurrent laryngeal nerve appear to be the cause of postoperative phonation disorders in patients undergoing esophagectomy for esophageal cancer, and these disorders can occur in the absence of symptoms such as hoarseness and dysphonia. PMID- 15532811 TI - Considerable multiple esophageal carcinoma. Implications of a new clinical entity. AB - BACKGROUND/AIMS: The multiple occurrence of primary squamous cell carcinoma of the esophagus is often observed, and most such occurrences are double cancers. There have also been some cases with three or more intra-esophageal cancers, however, no detailed clinicopathologic study has yet been performed in the literature. METHODOLOGY: Two hundred and fifty patients of primary esophageal squamous cell carcinoma without preoperative treatment that underwent esophageal resection were re-evaluated by serial histopathologic investigations and we analyzed the data of ten patients with three or more intraesophageal cancers. RESULTS: The clinical and histopathologic characteristics were as follows; 1) all but one of the cases were male, 2) all patients had a history of both heavy smoking and drinking but only one case had a family history of esophageal cancers among their siblings, 3) the depth of invasion in the carcinomas was restricted to within the submucosal layer of the esophageal wall, which was defined as superficial esophageal carcinoma, almost all (90%) of the cases accompanied esophageal squamous epithelial dysplasia. CONCLUSIONS: Based on these prominent characteristics of considerable multiple intra-esophageal cancers, a new clinical entity of "esophageal field cancers" could thus be suggested. PMID- 15532813 TI - Appraisal of the simultaneous right-thoracic and abdominal approach with intrathoracic reconstruction after esophageal resection for patients with intrathoracic esophageal cancer. AB - BACKGROUND/AIMS: This study evaluated the surgical procedure with the simultaneous right-thoracic and abdominal approaches with intrathoracic reconstruction after performing a macroscopically curable esophageal resection and a mediastio-abdominal extended lymphadenectomy as the standard operation for patients with intrathoracic esophageal cancer. METHODOLOGY: Forty-seven patients with thoracic esophageal cancer were operated on at the Department of Surgery I, Gunma University Hospital from 1995 to 1999. Before the end of 1997, all 21 cases underwent subtotal esophagectomy with three-field lymphadenectomy. After 1998, however, 16 cases underwent the same procedure while the remaining 10 underwent a simultaneous right-thoracic and abdominal approach with intrathoracic reconstruction with lymphadenectomy based on our criteria (Group I). Postoperative factors including the data of cytokines were compared between Group I and conventional subtotal (10 cases) esophagectomy with three-field lymphadenectomy (Group II, 37 cases). RESULTS: None of the patients in Group I has shown a recurrence of cancer. The CRP levels at 2 POD and the serum IL 6 levels at 3 POD were significantly lower in Group I than in Group II (p<0.05), and the postoperative duration of mechanical ventilation was also significantly shorter in Group I than in Group II (p<0.001). The postoperative body weight gain at 12 months after surgery was also higher in Group I than in Group II. CONCLUSIONS: This procedure is indicated for patients with esophageal cancer, located in the middle- or lower-thoracic esophagus, limited to within the esophageal wall, and without either upper mediastinal or neck lymph node metastasis, in order to not only cure the cancer but also in order to improve the postoperative quality of life. PMID- 15532814 TI - Long myotomy of the esophagus and gastric cardia with a complete fundic patch procedure for diffuse esophageal spasm. AB - Under the hypothesis that the surgical management of diffuse esophageal spasm requires the elimination or reduction of episodes of dysphagia and chest pain and prevention of postoperative gastroesophageal reflux, long esophageal myotomy and fundoplication had been performed. However, there have been some cases with unsatisfactory results. We describe herein a new surgical procedure of long myotomy of the esophagus and gastric cardia with a complete fundic patch operation for the patient with diffuse esophageal spasm. The advantages of this procedure are to preserve the separation of each myotomized edge and to reinforce the wall of the surface of the myotomized mucosa in order to avoid the postoperative problems. Postoperative course of the patient with this procedure was satisfactory. PMID- 15532815 TI - Evaluation of optimal gastric mucosal biopsy site and number for identification of Helicobacter pylori, gastric atrophy and intestinal metaplasia. AB - BACKGROUND/AIMS: The study is designed to identify the optimal gastric mucosal biopsy site and number for identification of Helicobacter pylori, gastric atrophy and intestinal metaplasia. METHODOLOGY: Ninety-two patients were included in the study, gastric biopsies were obtained from 5 different sites: lesser curvature of the mid-antrum (A1), greater curvature of the mid-antrum (A2), incisura angularis (IA), lesser curvature of the mid-corpus (B1), greater curvature of the mid corpus (B2). Helicobacter pylori was evaluated in sections stained with toluidine blue, and histopathological examination was performed in sections stained with hematoxylin-eosin. RESULTS: Seventy-three patients were positive for Helicobacter pylori at least in one biopsy site. Helicobacter pylori was positive in 47 patients (64.3%) in A1, in 54 patients (73.9%) in A2, in 60 patients (82.1%) in IA, 44 patients (60.2%) in B1, and in 42 patients (57.5%) in B2. The highest positivity determined was in the combination of A2 and IA sites (95.8%). Gastric atrophy was determined in 35 of 73 patients (27.1% in A1, 20% in A2, 25.7% in IA, 20% in B1, and 7% in B2). Intestinal metaplasia was determined in 31 of the Helicobacter pylori-positive patients (18% in A1, 16% in A2, 30.9% in IA, 21.8% in B1, 12.7% in B2). CONCLUSIONS: It is considered that taking biopsies from both A1 and IA sites has the highest sensitivity in detecting Helicobacter pylori. However, it is difficult to define a specific site for detecting gastric atrophy and intestinal metaplasia. PMID- 15532816 TI - Comparison of serum and urine ELISA methods for the diagnosis of Helicobacter pylori--a prospective pilot study. AB - BACKGROUND/AIMS: Helicobacter pylori infection has a close relationship with many upper digestive tract diseases, but a gold standard for identifying this infection has not yet been well established, especially in clinical application. There is an increasing interest in non-invasive diagnostic tests. In this study, we will compare two non-invasive tests (ELISA method of urine and serum) in clinical use. METHODOLOGY: 170 patients (82 women, 88 men; mean age 52.4 yr) were included in this study and all underwent gastroendoscopy. None of the patients had received any H. pylori eradication therapy. The H. pylori infection status was evaluated by histology, culture, CLO test, and 13C-UBT. We also collected urine and serum for the ELISA method. H. pylori infection was defined as positive if the culture was positive or if two of the other three tests (histology, CLO test and 13C-UBT) were positive. If all four tests were negative or only one of the other three tests was positive, the result would be interpreted as negative. RESULTS: Of these 170 patients, 107 (62.94%) were H. pylori infected and 63 (37.06%) were uninfected. The sensitivity, specificity, positive predictive value and negative predictive value of serum ELISA were 96.26%, 74.60%, 86.55%, and 92.16% respectively; while those of urine ELISA were 93.46%, 92.06%, 95.24% and 89.23% respectively. There was no significant difference between these two tests in sensitivity, but urine ELISA has better specificity than serum ELISA significantly. CONCLUSIONS: Urine ELISA is a better non-invasive test for H. pylori infection than serum ELISA in clinical practice, because no blood need be drawn, it is safer, easier, and more convenient for sampling and has better specificity. Therefore it would be suitable in epidemiological screening of H. pylori infection, especially in young children, older patients and those who cannot cooperate. PMID- 15532817 TI - A prospective within-patient comparison clinical trial on the effect of parenteral cimetidine for improvement of fluid secretion and electrolyte balance in patients with short bowel syndrome. AB - BACKGROUND/AIMS: Survival of patients with gastrointestinal cancer has improved recently with various multidisciplinary treatments. However, complications associated with the latter also increased, leading to impairment of quality of life and functional disability in patients. Short bowel syndrome, characterized by severe watery diarrhea and electrolyte losses, is one of the major problems caused by gastrointestinal cancer and subsequent treatments. We investigated whether cimetidine, a gastric antisecretory agent, would ameliorate short bowel syndrome in patients after massive small bowel resection. METHODOLOGY: The study design was such that the evaluation of cimetidine effects was an intra-patient comparison. The randomized block method was used for statistical analyses of frequency and amount of daily watery fluid excretion and urine output, electrolyte levels in fluid secretions, as well as blood biochemistry, body weight and performance status. RESULTS: Cimetidine administered at an early stage after bowel surgery, especially while patients are receiving parenteral nutrition, significantly decreased stool fluid excretion (p=0.0001), urine output (p=0.0010) and electrolyte loss, which was no longer apparent a week after initiation of oral intake. CONCLUSIONS: Cimetidine appears to be beneficial for cancer patients who have had extensive small bowel resection. PMID- 15532818 TI - Change in alpha glutathione s-transferase levels during liver resection. AB - BACKGROUND/AIMS: Alpha-glutathione s-transferase (GST) is a 50,000-kDa cytosol protein of the hepatocytes. It comprises 5% of the soluble protein of hepatocytes and is readily released in response to injury. Its half-life time is 60 minutes (AST: 47 hrs, ALT: 22 hrs). The aim of this study is to clarify the usefulness of GST measurement during liver resection. We obtained data from 26 patients undergoing liver resection and compared GST levels with AST and ALT levels. METHODOLOGY: Patients included 15 hepatocellular carcinoma, 5 cholangiocellular carcinoma and 6 metastatic cancers (4: colon, 2: stomach). We performed lobectomy for 11, segmentectomy for 5 and partial resection for 10 patients. From these patients, blood samples were collected before surgery, beginning of resection, end of resection, 15, 30, 60 min after resection, end of surgery, 1, 3, 6, 12, 24 hr after operation and 2, 3, 4, 5, 6, 7, 10, 14 postoperative days (POD). GST (Biotrin, Ireland) levels in the serum samples were determined by an ELISA. We also measured AST, ALT levels from the same set of samples. RESULTS: At the end of resection, GST showed highest level and quickly decreased at 1 hr after operation. On the other hand, AST and ALT showed peak levels at 12 hr after operation. CONCLUSIONS: Our findings suggest that perioperative GST measurement may be a sensitive and useful marker to evaluate liver function after liver resection. PMID- 15532819 TI - Prognostic factors for chemoembolization in liver metastasis from endocrine tumors. AB - BACKGROUND/AIMS: The aims of this study were to assess response rates, evaluate side effects and determine prognostic factors for both response and toxicity in patients with liver metastasis from endocrine tumors treated with chemoembolization. METHODOLOGY: Data concerning 64 patients who underwent a total of 186 sessions of chemoembolization were retrospectively evaluated and correlated with response and toxicity after chemoembolization. RESULTS: Overall clinical, morphological and biological response rates were 93%, 74% and 52% respectively. Complete control of hormone-related symptoms was obtained in 59% of patients with a mean duration of response of 15 months. Transient major complications occurred after 5.9% of sessions and 3 patients died. In the univariate and multivariate analyses, a non-pancreatic primary and chemoembolization as first-line non-surgical treatment were prognostic factors for clinical response, and <30% of liver involvement for morphological response. A significant increase in morbidity was noted in patients with more than 70% of liver involvement. CONCLUSIONS: Hormone-related symptoms were controlled in the majority of patients with a non-pancreatic primary and in those treated with chemoembolization as first-line therapy. Morphological response and toxicity were respectively correlated with liver involvement of less than 30% and greater than 70%. PMID- 15532820 TI - Surgical treatment of non-cirrhotic presinusoidal portal hypertension. AB - BACKGROUND/AIMS: Non-cirrhotic portal hypertension has a better prognosis than other forms of portal hypertension because of a well-preserved liver function in most cases. These patients are good candidates to receive surgical treatment, which is the therapeutic choice available with the lowest rebleeding rate. Because of abnormalities in the splanchnic vessels due to the nature of the diseases, many of them cannot be shunted. An extensive esophagogastric devascularization, the complete portoazygos disconnection, was evaluated. METHODOLOGY: A retrospective review of files of 31 patients, among 491 operations between 1991 an 2001 was carried out in a tertiary care Academic University Hospital. Patients comprised those with non-cirrhotic bleeding portal hypertension treated by means of complete portoazygos disconnection. Extensive two-stage (thoracic and abdominal) esophagogastric devascularization with modified transection of the esophagus was performed. MAIN OUTCOME MEASURES: recurrence of hemorrhage, encephalopathy and survival. RESULTS: Thirty-one patients were treated. In 17 cases (54%) a hypercoagulable state was demonstrated. No operative mortality was observed (0-30 days) with a total of 62 operations (two stages per patient). No case of encephalopathy was observed and in 3 cases (9%) rebleeding was recorded. The survival curve showed a 5-year survival of 97% and a 10-year survival of 93%. CONCLUSIONS: Complete portoazygos disconnection is an excellent surgical alternative for patients with non cirrhotic portal hypertension, with a low morbidity and mortality as well as a low rebleeding rate and good long-term survival. PMID- 15532821 TI - Factors in successful radiofrequency ablation therapy for malignant liver tumors. AB - BACKGROUND/AIMS: Radiofrequency ablation has become a new therapeutic method for treating malignant liver tumors. We reviewed our experience to identify the factors involved in successful radiofrequency ablation therapy. METHODOLOGY: Patients who underwent this therapy between 1999 July and 2002 July were reviewed for the characteristics of their tumors, clinical data and operative techniques used. RESULTS: Sixty-one patients (hepatocellular carcinoma 50 and metastatic tumors 11) were ablated. Forty-six cases (75.4%) were ablated effectively. Survival of patients with hepatocellular carcinoma was superior to those with metastases. The effective factor was the number of nodules while the survival factors were the number of nodules and the maximum tumor diameter. Recurrence factors in patients with hepatocellular carcinoma ablated effectively were poor hepatic function due to cirrhosis and higher protein induced by vitamin K absence or antagonist-II (PIVKA-II). CONCLUSIONS: The survival of patients with hepatocellular carcinoma was significantly better than those with metastases. We recommend radiofrequency ablation therapy for cases having a single hepatic tumor less than 3.5 cm in diameter (4 cm for hepatocellular carcinoma requiring much care). In patients with hepatocellular carcinoma, hepatic function with cirrhosis and PIVKA-II showed a significant correlation with recurrence. PMID- 15532822 TI - Low mortality major hepatectomy. AB - BACKGROUND/AIMS: A retrospective series of major hepatectomy using a simple strategy. METHODOLOGY: From 1995 to 2002, 104 major hepatectomies were performed, including 16 extended and 35 right/left lobectomies, 11 bisegmentectomies, and 42 left lateral segmentectomies. Seventy-two patients had malignant tumors. Chronic hepatitis was present in 9 and mild to severe cirrhosis in 16 of 47 hepatocellular carcinomas. Intrahepatic duct stone constituted 28 of 32 patients with benign disease. Hilar ligation without pedicle clamp for lobectomy and Pringle's maneuver for segmentectomy were initially our standard procedures. Since 1997, Chang's needle was applied on 34 patients to block hepatic inflow and outflow by entire thickness, interlocking mattress sutures. RESULTS: Three patients (2.9%) died postoperatively. Of the two cholangiocarcinoma patients, one underwent extended right lobectomy and died of multiple organ failure and the other underwent left lateral segmentectomy and died of AMI unrelated to the surgery. One hepatocellular carcinoma patient underwent left lateral segmentectomy and died of hepatic failure. Significant complications (11.5%) included three bile leaks (2.9%), six abscesses (5.8%), one hepatic failure (1%), one ileus (1%), and one ventral hernia (1%). One major bile leak necessitated a reconstructive surgery. CONCLUSIONS: Our strategies for hepatic resection are simple, however we are able to achieve a low mortality major hepatectomy. PMID- 15532823 TI - The effects of a bradykinin B2 receptor antagonist in extended liver resection with ischemia in dogs. AB - BACKGROUND/AIMS: In liver surgery, total clamping of the portal triad (Pringle's procedure) is commonly used, and sometimes causes liver failure. This study evaluated the effects of a bradykinin B2 receptor antagonist, FR173657 (FR), on ischemia-reperfusion injury during liver resection in dogs. METHODOLOGY: Experimental animals were divided into two groups. In the FR group (n=6), FR (100 nmol/kg/hr) was administered continuously via the portal vein from 30 min before the onset of ischemia until 2 hr after reperfusion. In the control group (n=6), vehicle was injected in the same manner. The right portal pedicle was clamped for 60 min, while the left portal branch was left patent to avoid portal congestion. Following reperfusion, the non-ischemic lobes were resected, and remnant liver function was evaluated. RESULTS: AST and ALT were significantly (p<0.05) lower in the FR group than in the control group. Hepatic tissue blood flow 30 min after reperfusion was significantly (p<0.05) higher in the FR group than in the control group. Histological tissue damage was mild, and polymorphonuclear neutrophil infiltration was significantly (p<0.05) reduced in the FR group compared with the control group. CONCLUSIONS: A bradykinin B2 receptor antagonist ameliorated the ischemia-reperfusion injury caused by Pringle's procedure during extended liver resection. PMID- 15532824 TI - Preoperative hyaluronic acid measurement to assess the hepatic functional reserve. AB - BACKGROUND/AIMS: With liver surgery, preoperative assessment of the hepatic functional reserve is important for estimating the extent of hepatectomy. Hyaluronic acid (HA) is specifically cleared and metabolized by endothelial cells in the liver. Its half-life time is 2-5 min and HA might be a sensitive liver function marker. We obtained data from 44 patients with liver tumor undergoing liver resection. METHODOLOGY: We studied 44 liver resected patients. Blood samples were obtained before surgery and HA levels and other liver function markers [type IV collagen (IV), procollagen-III-peptide (PIIIP), lecithin cholesterol acyltransferase (LCAT), cholinesterase (ChE), albumin (Alb), platelets (Plt), prothrombin time (PT%)] levels in the samples were measured. We also performed indocyanine green retention test and calculate R15% (ICG). RESULTS: HA showed strong positive correlation with ICG, IV and PIIIP, negative correlation with LCAT, ChE, Alb, Plt and PT%. ICG was not correlated with ChE, Plt and PT%. The HA levels and ICG of the patients who had more than 2 segments of the liver resected were below 100 ng/mL and 20%, respectively. In the patients with obstructive jaundice HA levels of the 3 patients with obstructive jaundice who underwent bisegmentectomy were below 100 ng/mL. CONCLUSIONS: Our findings suggest that HA is a useful preoperative liver function test as well as ICGR15%. Preoperative HA levels <100 ng/mL and ICGR15% <20% might be helpful indicators for major liver resection. We also recommend that HA measurement for evaluating liver function in the patients with obstructive jaundice ICG is not helpful. PMID- 15532825 TI - Serum level of transforming growth factor-beta1 (TGF-beta1) and the expression of TGF-beta receptor type II in peripheral blood mononuclear cells in patients with autoimmune hepatitis. AB - BACKGROUND/AIMS: To evaluate the association of the immunosuppressive effects of transforming growth factor-beta1 (TGF-beta1) with abnormalities in immune regulation in autoimmune hepatitis (AIH), we investigated the serum level of TGF beta1 and expression of TGF-beta receptor type II (TbetaRII) in peripheral blood mononuclear cells (PBMC) in patients with AIH. METHODOLOGY: Twenty-two patients with AIH were included in this study. Serum levels of total TGF-beta1 were determined using a specific enzyme-linked immunosorbent assay (ELISA). The expression levels of TbetaRII mRNA were semi-quantitatively determined by ribonuclease (RNase) protection assay specific for TbetaRII. RESULTS: The mean serum level of TGF-beta1 in patients with AIH (230+/-95 ng/mL) was higher than that of healthy controls (137+/-81 ng/mL, p=0.012). The expression level of TbetaRII mRNA in PBMC obtained from AIH patients (0.131+/-0.046) was lower than that in PBMC of patients with chronic hepatitis C (0.186+/-0.074, p=0.019) and that of healthy subjects (0.188+/-0.060, p=0.013). CONCLUSIONS: The present study of TbetaRII mRNA expression in PBMC from patients with AIH suggested that the decreased expression of TbetaRII might contribute partially to abnormalities in immune regulation observed in patients with AIH, despite concomitant up regulation of TGF-beta1 production. PMID- 15532826 TI - Microwave coagulation therapy for unresectable hepatocellular carcinoma. AB - BACKGROUND/AIMS: Surgical resection is not always feasible for patients with hepatocellular carcinoma. We used microwave coagulation therapy (MCT) as an alternative to resection and evaluated its efficacy. METHODOLOGY: Twenty-four patients with unresectable hepatocellular carcinoma underwent microwave coagulation therapy by laparotomy (n=18), laparoscopy (n=4), or thoracoscopy (n=2) because of advanced liver cirrhosis and/or intrahepatic metastases. One nodule was treated in 11 patients, 2 nodules were treated in 7, 3 nodules were treated in 3, 6 nodules were treated in 1, and 7 nodules were treated in 2. Tumor size ranged from 10 to 50 mm. Liver function was analyzed at the time of initial MCT and at treatment for recurrence. Patient outcomes were studied. RESULTS: Two patients died postoperatively after initial MCT. Other patients showed rapid recovery without hepatic dysfunction. Liver function just before MCT was equivalent to that measured just before treatment for recurrence. One patient developed local recurrence at the margin of the treated tumor. Recurrent nodules in different segments were detected in 15 patients. Transcatheter arterial embolization was performed in 13 recurrences, percutaneous ethanol injection therapy was performed in 1 recurrence, and MCT was performed in 1 recurrence. The 3-year cancer-free survival rate was 9.9%, and the 3-year cumulative survival rate was 83.9%. CONCLUSIONS: Because MCT is indicated for hepatocellular carcinoma patients with advanced liver cirrhosis, intrahepatic recurrences are frequent. Since, liver function is preserved after MCT, however, locoregional therapy can be selected when intrahepatic recurrence is detected, thus improving patient survival rate. PMID- 15532827 TI - Simultaneous percutaneous right portal vein embolization and left liver tumor radiofrequency ablation prior to a major right hepatic resection for bilateral colorectal metastases. AB - BACKGROUND/AIMS: Hepatic resection offers the best chance of survival for patients with liver metastases (LM) of colorectal origin. However, some patients are not eligible for surgery because of a too small future liver remnant (FLR) which carries a high risk of severe postoperative liver failure. The operability status of these patients can be favorably changed by selective right portal vein embolization (PVE) which induces compensatory growth of the left liver. However, during liver regeneration following right PVE, the left LM growth rate is faster than that of the non-embolized normal liver parenchyma. This study aimed at examining an approach for those patients in which there is bilateral LM potentially resectable following portal vein embolization, but in which there is a risk of rapid liver metastasis growth in the non-embolized liver. METHODOLOGY: Between October 1998 and January 2001, 5 patients underwent simultaneous right PVE and radiofrequency ablation (RFA) of a left LM, prior to a major right-sided hepatectomy for initially unresectable bilateral LM. All these patients had one LM in the left liver in addition with multiple LM in the right liver. Simultaneous right PVE and left RFA was performed percutaneously under intravenous sedation and analgesia. One month later, hepatectomy was undertaken. To allow histologic assessment of the RFA effectiveness, the previously treated left-sided tumor was also resected and analyzed. RESULTS: Simultaneous PVE-RFA was successful in all patients. No tumor growth on the RFA site was observed during the interval between PVE-RFA and surgery. Histologic examination showed complete tumor sterilization of the RFA necrotic zone. In the postoperative course, 1 patient died of acute liver failure. For the 4 remaining patients, morbidity was minimal (transient bile leak in one patient). CONCLUSIONS: Simultaneous percutaneous right PVE and left RFA is feasible. This procedure allowed good left-sided tumor control during liver growth following PVE in all five patients. It is the most logical procedure for patients with bilateral colorectal LM needing right PVE before resection, if the left concomitant LM is small and accessible to percutaneous RFA. This procedure should be preferred because it eliminates the risk of left LM growth during the 1-month interval between PVE and surgery. PMID- 15532828 TI - Risk factors for postoperative liver failure after hepatectomy for hepatocellular carcinoma. AB - BACKGROUND/AIMS: Selection of patients for hepatectomy for hepatocellular carcinoma conventionally has been based upon Child-Pugh grading. However, postoperative liver failure after hepatectomy is a major cause of hospital mortality. A new predictor of postoperative liver failure is required. The objective of this study was to identify risk factors for postoperative liver failure after hepatectomy. METHODOLOGY: Perioperative risk factors for liver failure after hepatectomy were analyzed in 112 patients with hepatocellular carcinoma Eight of these patients died of liver failure. Stepwise multivariate logistic regression was performed to investigate significant independent factors among 17 variables, including the serum alkaline phosphatase ratio (ALPR) on the first day after hepatectomy. ALPR was calculated as the postoperative ALP level divided by the ALP level before surgery. RESULTS: Significant risk factors of postoperative liver failure were ALPR on postoperative day 1 (ALPR1), sex, operative blood loss, and operative procedure. As an indicator of liver failure, the diagnostic accuracy of the ALPR1 was 93.7% when the ALPR was less than 0.4 on the first postoperative day. The ALPR and the serum total bilirubin concentration after hepatectomy were uncorrelated. CONCLUSIONS: ALPR1 is a useful predictor of liver failure after hepatectomy. PMID- 15532829 TI - Effects of posture change on the hemodynamics of the liver. AB - BACKGROUND/AIMS: According to our experience, blood flow in the portal vein may alter according to body posture. It is reported that decreased portal venous flow immediately gives rise to significantly increased blood flow in the hepatic artery. To gain further insight into blood flow changes affected by posture, we examined blood flows in the portal vein, hepatic artery and hepatic vein at different postures. METHODOLOGY: Using a Doppler ultrasound system, the hemodynamics of the portal vein, right hepatic artery, and hepatic vein were examined in 35 patients at supine and left decubitus positions. RESULTS: Portal vein blood flow volumes were significantly lower in the left decubitus position than in the supine. In the right hepatic artery, the left decubitus position gave significantly higher blood flow velocity values than the supine. CONCLUSIONS: Our results indicated that upon change of posture from the supine to left decubitus position, portal vein flow velocity was reduced and hepatic artery flow velocity increased. Changes in portal and hepatic arterial flows by changing posture may be explained by decreased portal flow as a direct result of changed posture, leading to increased hepatic arterial flow to maintain total hepatic blood inflow. PMID- 15532830 TI - ERCP in orthotopic liver transplanted patients. AB - BACKGROUND/AIMS: Biliary complications after orthotopic liver transplantation (OLT) are still common. The aim of the study was to assess the effectiveness of ERCP as a diagnostic and treatment tool in the management of biliary tract OLT complications. METHODOLOGY: The diagnostic and treatment effectiveness of ERCP in the management of biliary tract OLT complication from 1/1995 to 12/2001 was reviewed. RESULTS: 24/251 (9.6%) OLT patients presented biliary complications. These patients underwent a total of 31 ERCP (seven patients received two ERCP). The ERCP indication was cholestasis in 25 (80.6%). The success rate of these 31 ERCPs was 87%. In the 25 ERCPs indicated in patients with cholestasis, the procedure revealed strictures of the biliary anastomosis in 3, hepatic hilum strictures in 5, SOD in 4, lithiasis in 7 and sclerosing cholangitis in one. The final diagnosis of three patients with normal biliary tract was intrahepatic cholestasis. In three of the four patients with biliary leaks the ERCP's confirmed the diagnosis. The final treatment efficacy was 100% (7/7 patients), 50% (2/4 p.), 0% (0/3 p.), 75% (3/4 p.) and 100% (4/4 p.) for patients with biliary stones, strictures of the hilum, anastomotic strictures, biliary leaks and SOD, respectively. In the acute pancreatitis and in the PSC endoscopic treatment was not indicated. There were two mild cases of pancreatitis. CONCLUSIONS: Because of its availability, diagnostic accuracy, treatment efficacy and safety, ERCP should be used as the first-line procedure for treatment of biliary tract complications after OLT. PMID- 15532831 TI - Surgical treatment for extrahepatic recurrence after hepatectomy for colorectal metastases. AB - BACKGROUND/AIMS: Recurrence of the remnant liver or extrahepatic sites after hepatic resection for colorectal metastases is concerned as a significant prognostic factor. While regional chemotherapy may reduce recurrence of the liver, appropriate therapy for extrahepatic metastases needs to be elucidated. METHODOLOGY: We identified 207 patients undergoing hepatectomy for colorectal metastases. Patient characteristics, clinicopathological features, sites of extrahepatic recurrence, and outcome are examined. RESULTS: Recurrence of the lung, brain, bone, and other site after hepatic resection was seen in 49, 8, 8, and 12 cases, respectively. Fourteen patients underwent pulmonary resection, and three patients are alive and disease-free survival was 35, 79, and 128 months after the second resection. Overall 2- and 5-year survival was 64 and 18%. Univariate and multivariate analysis indicated that time and number of pulmonary metastases, and serum CEA levels are independent prognostic factors. Surgical resection for brain metastasis was only in one case who survived for 31 months. Other treatment such as gamma-knife may not improve outcome. CONCLUSIONS: Both hepatic and pulmonary resection for colorectal metastases could prolong survival in selected patients. Surgical indication for other extrahepatic metastases must be strictly selected, although resection of brain metastasis might lead to better outcome. PMID- 15532832 TI - Surgical treatment for liver metastases from colorectal carcinoma: results of 228 patients. AB - BACKGROUND/AIMS: Liver metastases are a very common event. Multiple choices of therapies can be used. The aim of this paper is to analyze results and methods of a single institution series of 228 consecutive patients with colorectal liver metastases. METHODOLOGY: 228 consecutive patients underwent hepatic resection for colorectal liver metastases. From different periods intraoperative ultrasound, intraoperative histological examination, locoregional intra-arterial chemotherapy, and radiofrequency thermal ablation were introduced. RESULTS: Operative mortality was 0.9%. Mean follow-up was 29.5 months. Overall survival was 16% and 9% at 5 and 10 years. 5-year survival was 23% and 6% for patients with single and multiple metastases respectively. For patients with extrahepatic metastatic single lesion 5-year survival was 15%. From the start of intraoperative ultrasound use, 5-year survival was 9% and 27% for patients with multiple and single metastases. Five-year survival for re-resected patients was 13%. Overall survival at 1 and 3 years was 90% and 58% in patients treated with HAI and systemic chemotherapy (disease-free 70% and 47%) and 94% and 12% in patients treated with systemic chemotherapy alone after radical resection (disease-free 53% and 0%). CONCLUSIONS: Aggressive approach, re-resections, intraoperative ultrasound staging, intra-arterial chemotherapy and radiofrequency thermal ablation are justified in multimodal therapeutic strategy of colorectal metastases and seem to improve patients' survival. PMID- 15532833 TI - Acute renal failure during the early postoperative period in adult living-related donor liver transplantation. AB - BACKGROUND/AIMS: Acute renal failure after liver transplantation can occur in some and is an important postoperative complication. Our goal is to clarify the risk factors of acute renal failure after living-related donor liver transplantation (LDLT). METHODOLOGY: From March 1999 to August 2000, ten consecutive patients were investigated the changes of the systemic hemodynamics and the renal function. They were classified into Group A (Creatinine (Cre) was over 2.0 mg/dL) and B (Cre was below 2.0 mg/dL). Retrospective variables were examined with two groups A and B being compared. RESULTS: In both groups, Cardiac Index (CI) was above standard levels. However, the CI levels in Group B were significantly higher than those in Group A (p=0.031). The early postoperative transaminase levels were significantly higher in Group A than in Group B (p=0.049) and graft liver volume/recipient body weight ratio was significantly smaller in Group A than in Group B (p=0.016). CONCLUSIONS: Our study suggests that small-for-size graft or hypovolemia, resulting in the delay of the recovery of graft liver function, may be an important cause of acute renal failure during the early postoperative period in adult LDLT. PMID- 15532834 TI - Changes in hepatic lobe volume in hepatocellular carcinoma after transcatheter arterial and percutaneous transhepatic portal embolization. AB - BACKGROUND/AIMS: Determining changes in liver volume after preoperative percutaneous transhepatic portal embolization (PTPE) for hepatocellular carcinoma (HCC) is essential in managing the operation safely. We evaluated the alterations in liver volume by means of ultrasonography (US) and computed tomography (CT). METHODOLOGY: We studied 12 patients scheduled for hepatectomy with HCC. Transcatheter arterial embolization (TAE) and PTPE of the right portal vein were performed preoperatively. Liver volume was evaluated before and after PTPE. RESULTS: The volume of the right lobe measured by CT significantly decreased from 709+/-266 cm3 before PTPE to 589+/-209 cm3 18 days after PTPE (P=0.0021). The volume of the left lobe significantly increased from 382+/-97 cm3 to 477+/-84 cm3 (P=0.0008). US volume measurement of the left lateral segment revealed a temporary volume increase 2 days after PTPE and a significant linear correlation between the hypertrophy ratios from 2 to 18 days after PTPE (r=0.946, P<0.0001). CONCLUSIONS: Preoperative PTPE allowed a compensatory volume increase in the remnant liver. A sonographic estimation is useful in confirming the dynamic alteration of liver hypertrophy. Care must be taken for appropriate timing of a CT scan for volume measurements, considering the drawbacks associated with irradiation. PMID- 15532835 TI - Gastrointestinal bleeding during liver transplantation--report of two cases. AB - A few authors have reported, especially as intraoperative complications, gastrointestinal hemorrhage related to liver transplantation. The aim of this study was to show two cases of gastrointestinal hemorrhage, which occurred during surgery. The first patient was male, 46 years old, with viral hepatic cirrhosis. He had previously presented two episodes of digestive bleeding. Upper digestive endoscopy showed esophageal gastric varices. During the hepatectomy there was bleeding inside the nasogastric tube associated with severe hemodynamics instability without other sources of bleeding. Intraoperative endoscopy evidenced bleeding gastric varices. Gastrectomy was carried out and the varices were tied. The piggyback technique was used in the liver transplantation. The surgery was concluded without problems and in the following four and a half years his condition has evolved well. In the second case, the patient was aged 17, female, with autoimmune hepatic cirrhosis. She had previously presented one episode of digestive bleeding. Intraoperative endoscopy showed median esophageal varices. During the anesthetic induction she presented an episode of hematemesis. A Sengstaken-Blakemore balloon was introduced. The transplant was performed without further problems. Her case has been followed for 14 months in the outpatients' clinic with a good postoperative course. To sum up, gastrointestinal hemorrhage can be due to portal hypertension during the liver transplantation and must be treated quickly. In these cases the surgery must be ongoing. PMID- 15532836 TI - Is peritoneal cytology a predictor of unresectability in pancreatic carcinoma? AB - BACKGROUND/AIMS: Demonstration of unresectability often requires diagnostic laparotomy with high co-morbidity in pancreatic carcinoma. The value of detected peritoneal tumor cells and influence on outcome should be determined. METHODOLOGY: In a prospective study 150 consecutive patients with pancreatic carcinoma were evaluated. Improvement of diagnostic accuracy in diagnosing unresectability was calculated for combination of CT, endoscopic ultrasonography and peritoneal cytology. RESULTS: Unexpected subglissonian metastases were found in 10%. 87 patients with peritoneal washings were included in the study, 20 patients with detected peritoneal tumor cells (n=22) were inoperable. In all patients of this group the ventral integrity of pancreas was damaged. In 23 patients with preoperative existent ascites, only in 4 cases could peritoneal tumor cells be detected. In patients with positive cytology together with disrupted ventral pancreatic margin as predictors of unresectability, sensitivity was 100% and specificity 96%. CONCLUSIONS: Addition of diagnostic laparoscopy combined with peritoneal cytology and minimally invasive palliation procedures (endoscopic biliary prosthesis, laparoscopic gastrojejunostomy) to the staging protocol for pancreatic carcinoma may decrease operative lethality and unnecessary laparotomies may be avoided safely. As unexpected subglissonian micrometastases may additionally be present in 5-10%, perioperative laparoscopy is always advisable. PMID- 15532837 TI - Cyclin D1 and bax influence the prognosis after pancreatoduodenectomy for periampullary adenocarcinoma. AB - BACKGROUND/AIMS: The outcome of patients undergoing pancreatoduodenectomy for periampullary adenocarcinoma is influenced by various clinicopathological factors, chemo/radiotherapy and expression of various oncogenes, tumor suppressor genes, growth factors and factors controlling apoptosis. The aim of this study was to define clinicopathological predictors of survival in periampullary adenocarcinoma, and to determine the prognostic significance of cyclin D1, p27kip1 and bax expression in these tumors. METHODOLOGY: Prospectively, we collected clinicopathological data for patients operated on between January 1995 and December 1998. Cyclin D1, p27kip1 and bax expression was assessed immunohistochemically. The potential influence of clinicopathological factors and cyclin D1, p27kip1 and bax expression on survival was investigated. Univariate analyses were performed using the Kaplan-Meier method and log-rank test. For multivariate analysis Cox proportional hazards regression was used. RESULTS: Fifty-five patients were included in the study. The actuarial 5-year survival was 30%: 11% for pancreatic adenocarcinoma and 46% for distal bile duct/papillary adenocarcinoma Univariate analysis identified diabetes mellitus, blood transfusion, diameter of the tumor, histological type of the tumor, lymphatic invasion, neural invasion, lymph node metastasis, overexpression of cyclin D1 and lower expression of bax and p27kip1 as factors that significantly decrease survival rates. In multivariate analysis, the histological type of the tumor (p=0.001), lymph node involvement (p=0.03) and overexpression of cyclin D1 (p=0.02) independently influenced survival, whereas decreased expression of bax nearly reached statistical significance (p=0.054). CONCLUSIONS: Our findings confirm the association between cyclin D1 and bax expression and aggressive biological behavior of periampullary adenocarcinomas. Moreover, these parameters were identified as independent prognostic indicators in these tumors. PMID- 15532838 TI - Postoperative pancreatic fistula following distal pancreatectomy for pancreatic neoplasm; can pancreatic fistula be prevented? AB - BACKGROUND/AIMS: Pancreatic fistula is a potentially fatal complication in pancreatic surgery. The objective of this study was to analyze the incidence of pancreatic fistula in patients who underwent distal pancreatectomy for pancreatic neoplasm. METHODOLOGY: Forty-seven patients who underwent distal pancreatectomy for neoplasm of the pancreas at Kochi Medical School between October 1981 and December 2002 were studied. Comparative analysis was carried out to identify the correlations between the incidence of pancreatic fistula and parameters of preoperative evaluation, intraoperative procedure, and postoperative laboratory investigation. RESULTS: The incidence of pancreatic fistula in patients who underwent distal pancreatectomy for pancreatic neoplasm was 27.7%. Intraoperatively, the frequency of pancreatic fistula was significantly high in patients who underwent distal pancreatectomy without lymph node dissection and ligation of the splenic artery. No significant difference was recognized between the conventional surgical division and auto suture staple groups, however, in the ultrasonically activated scalpel (USAS) group, no pancreatic fistula was observed. A multivariate analysis revealed that independent high incidence of pancreatic fistula were 1) the situation of the tumor at pancreatic body, 2) the procedure of D2 lymph node dissection, and 3) the ligation of the root of splenic artery. In the group with the presence of pancreatic fistula, the serum amylase level on the first postoperative day was significantly higher than that in the group without fistula. CONCLUSIONS: Our findings supported that pancreatic fistula may be preventable by use of the USAS and treatment of acute pancreatitis in the early postoperative stage. PMID- 15532839 TI - Sequential and/or concurrent hypofractionated radiotherapy and concurrent chemotherapy in neoadjuvant treatment of advanced adenocarcinoma of the pancreas. Outcome and patterns of failure. AB - BACKGROUND/AIMS: To investigate treatment outcome and patterns of failure of sequential chemotherapy (CHT) and/or concurrent hypofractionated radiotherapy (RT) and CHT followed by surgery in locally advanced non-metastatic pancreatic adenocarcinoma. METHODOLOGY: Seven patients with locally advanced but marginal resectable tumors (close contact but no signs of infiltration of the mesenteric vessels and/or vena portae) were treated with hypofractionated RT (5x3 Gy per week) and concurrent continuous infusion (300 mg/sqm/24 h, 7 days per week) of 5 fluorouracil (FU). Ten patients with locally advanced disease with radiologically suspected infiltration of the mesenteric vessels and/or v. portae were treated with 2 cycles of Cisplatin (75 mg/sqm) and Gemcitabine (2x1250 mg/sqm), and patients without tumor progression received the same concurrent RT/CHT as group 1. Four weeks after RT/CHT radical pancreatectomy was planned for patients with stable disease or remission. RESULTS: Toxicity was low in both groups, with no CTC grade 4 toxicity. In group 1, RT/CHT was completed in all patients. There was no radiological remission, but stable disease in 5 out of 7 patients. All 5 patients underwent resection of the primary tumor with a R0-resection in 3 patients. In group 2, 8 patients completed CHT and RT/CHT treatment as planned. There were 3 with partial remission. Operation was done in 4 patients, but only one R0 resection was achieved. The median survival time for all 17 patients is 13 months, with 1- and 2-year survival being 53% and 18%, respectively. Local progression was observed in 9, peritoneal seeding in 7 and distant metastasis (mostly liver and lung) in 8 patients. CONCLUSIONS: The neoadjuvant therapy could be administered with low toxicity. Results of this study warrant further investigation aiming at optimal tailoring in of this treatment approach in these two subgroups of patients. PMID- 15532840 TI - Arterial hemorrhage by pseudoaneurysms following pancreatoduodenectomy. AB - Experienced centers report reduced operative mortality after pancreatoduodenectomy for cancer, but significant complications continue to occur in as many as 25% of patients. Anastomotic disruption leads to sepsis in the pancreatic bed where major vascular structures have been exposed by regional lymphadenectomy, creating a setting for arterial erosion or ligature slough and massive hemorrhage. Dehiscence of the pancreatojejunostomy is a particular risk. We presented one patient with arterial hemorrhage from visceral artery pseudoaneurysm after pancreatoduodenectomy, who had had sentinel bleeding from the abdominal drains or gastrointestinal tract. Sentinel bleeding after pancreatoduodenectomy indicates local sepsis and probable anastomotic dehiscence. Recognition of its significance and prompt response may prevent exsanguination. PMID- 15532841 TI - Curative resection for a pancreatic endocrine carcinoma involving the portal vein. AB - A 68-year-old male was admitted to our hospital for worsening of diabetes mellitus. Abdominal computed tomography showed a well-enhanced tumor in the head of the pancreas. Endoscopic retrograde cholangiopancreatography revealed tapering obstruction of the main pancreatic duct and stricture of the common bile duct. Celiac angiography demonstrated a vague tumor stain. Percutaneous transhepatic portography disclosed stricture of the portal vein. Pylorus-preserving pancreatoduodenectomy with segmental resection and reconstruction of the portal vein was performed under a diagnosis of nonfunctioning endocrine carcinoma of the pancreas. Histological examination demonstrated that the tumor was composed of small nests and cords of cuboidal cells with nodal invasion, and was proven to be nonfunctioning endocrine carcinoma of the pancreas. Microscopically, the tumor invaded the portal vein, common bile duct and main pancreatic duct. The postoperative course was uneventful, and he is now well without any evidence of recurrent disease 14 months after surgery. PMID- 15532842 TI - Accelerated rehabilitation with early postoperative oral feeding following gastrectomy. AB - BACKGROUND/AIMS: To evaluate the safety and efficacy of early oral feeding in patients undergoing gastrectomy. METHODOLOGY: One hundred patients undergoing gastrectomy were studied. Patients in the early oral feeding group (Early group) began a liquid diet within 48 hours after operation and patients within the Traditional group received nothing by mouth until the resolution of the ileus. All of the patients were monitored for vomiting, abdominal distention, length of ileus, tolerance of regular diet, duration of intravenous fluid administration, length of hospitalization, and complications. RESULTS: The time to flatus was 55.5+/-12.5 hours and 78.0+/-22.2 hours in the Early and Traditional group, respectively (p<0.05). And fasting period was 2.14+/-1.08 days and 5.93+/-2.35 days in the Early and Traditional group, respectively (p<0.05). In addition, duration of intravenous fluid administration was shorter in the patients in the Early group compared with Traditional group (5.7+/-1.7 days vs. 9.2+/-3.9 days, p<0.05). As a result, length of postoperative hospitalization in the patients in the Early group was significantly shorter than those in the Traditional group (16.2+/-5.3 days vs. 23.4+/-9.8 days, p<0.05). The incidence of complications including nausea, vomiting, anastomotic leak and wound infection occurred equally in both groups. CONCLUSIONS: Early oral feeding after gastrectomy is safe, with no evidence of increased morbidity, and early postoperative oral feeding is also highly effective in reducing hospital stay. PMID- 15532843 TI - Effects of mineral-water supplementation on gastric emptying of solids in patients with functional dyspepsia assessed with the 13C-octanoic-acid breath test. AB - BACKGROUND/AIMS: Functional dyspepsia is a major problem in terms of prevalence and drug-therapy expenditures. In dyspeptic patients the symptoms are frequently caused by delayed gastric emptying. Conventional treatment is often inefficient. Mineral-water supplementation is prescribed for the treatment of this condition, but there is no concrete proof of its actual efficacy. The aim of this study was to evaluate the effect of supplementation with mineral waters with high mineral salt contents (Acqua Tettuccio, Acqua Regina, Montecatini Terme) on gastric emptying of solids and symptoms in patients with functional dyspepsia. METHODOLOGY: Eight patients with functional dyspepsia and two healthy (non dyspeptic) controls were placed on high-mineral-content water supplementation (500 cc/day) for eight days. Before and after completion of the supplementation treatment, patient symptoms were scored and the 13C-octanoic-acid breath test was administered to assess gastric emptying. RESULTS: After the treatment, the dyspeptic subjects presented clear decreases in parameters of gastric emptying (half time and lag time) as well as an improvement in symptom scores. CONCLUSIONS: Health spa treatments based on consumption of waters with a high content of mineral salts seem to be capable of improving gastric emptying of solids in dyspeptics. Longer and longitudinal studies are needed to verify the persistence of this effect. PMID- 15532844 TI - Preoperative plasma fibrinogen levels in gastric cancer patients correlate with extent of tumor. AB - BACKGROUND/AIMS: The aim of the present study was to investigate the relationship between the preoperative plasma fibrinogen level and the extent of tumor involvement in gastric cancer patients. METHODOLOGY: Preoperative plasma fibrinogen levels of 354 patients who underwent gastric cancer surgery were quantified using an immunoassay. The relationships between the plasma fibrinogen level and other prognostic variables (tumor size, macroscopic and histological type, depth of tumor invasion, presence of lymph node involvement and distant metastasis) were then examined using univariate and multivariate linear regression analyses. RESULTS: The plasma fibrinogen level was significantly lower in patients with early gastric cancer than in those with advanced gastric cancer (312+/-6.7 vs. 361.9+/-97.0 mg/mL, p<0.001). A significant relationship existed between the preoperative fibrinogen levels and the presence of metastatic lymph nodes (320+/-78.6 vs. 352.6+/-94.1 mg/mL, p=0.001) and distant metastasis (338.2+/-89.5 vs. 396.9+/-128.3 mg/mL, p=0.013). Size of the tumors and depth of tumor invasion could predict elevated fibrinogen levels positively in both the univariate regression and multivariate linear regression analyses. CONCLUSIONS: These data suggest that the plasma fibrinogen level is a clinically important and useful marker of the extent of tumor progression in gastric cancer. PMID- 15532845 TI - Evaluation of P-POSSUM and O-POSSUM scores in patients with gastric cancer undergoing resection. AB - BACKGROUND/AIMS: The aim of this study was to evaluate the predictive accuracy of P-POSSUM and O-POSSUM models on patients undergoing elective gastric resection. METHODOLOGY: P-POSSUM and O-POSSUM predictor equations for mortality were applied retrospectively to 126 patients who had undergone elective gastrectomy for cancer. Observed mortality rates were compared with rates predicted by P-POSSUM and O-POSSUM using the Hosmer-Lemeshow goodness-of-fit test. Evaluation of the discriminative capability of both models was performed using receiver-operating characteristic (ROC) curve analysis. RESULTS: Overall fourteen deaths were observed. O-POSSUM predicted 15 deaths (chi2=14.61, p=0.13) and P-POSSUM predicted 20 deaths (chi2=25.41, p=0.002) using the Hosmer-Lemeshow test. ROC curves analysis revealed that O-POSSUM had better discriminatory power for mortality compared to P-POSSUM (area under curve=0.880, for O-POSSUM and area under curve=0.703 for P-POSSUM). CONCLUSIONS: These data suggest that O-POSSUM predicts mortality more accurately than P-POSSUM in patients undergoing elective gastrectomy for cancer. PMID- 15532846 TI - Role of pyloroplasty after proximal gastrectomy for cancer. AB - BACKGROUND/AIMS: The necessity of pyloroplasty as a drainage procedure after proximal gastrectomy remains controversial in terms of the postoperative quality of life. The aim of this retrospective study was to evaluate whether pyloroplasty is necessary after proximal gastrectomy. METHODOLOGY: Thirty-five patients who underwent proximal gastrectomy with jejunal interposition between 1993 and 2000 were studied. They were divided into two groups: Group A, jejunal interposition without pyloroplasty (n=17); Group B, jejunal interposition with pyloroplasty (n=18). The subjects were interviewed and examined to assess their symptoms, food intake and body weight at 6, 12, and 24 months after the operation. Endoscopy and a radioisotope gastric emptying test were performed one year postoperatively. RESULTS: A low incidence of epigastric fullness, nausea, and vomiting and a high frequency of patients with greater than 80% of pre-illness food intake were found in Group B. A high recovery of bodyweight was also achieved in Group B. On the other hand, a high incidence of reflux gastritis or bile regurgitation was found in Group B. Gastric emptying was significantly delayed in Group A. CONCLUSIONS: Pyloroplasty as a drainage procedure after proximal gastrectomy is necessary in terms of the clinical symptoms, dietary intake, recovery of body weight, or gastric emptying. PMID- 15532847 TI - Phase-II randomized study of preoperative IL-2 administration in radically operable gastric cancer patients. AB - BACKGROUND/AIMS: Surgery has appeared to induce lymphocytopenia and this decrease in host defenses during postoperative period could promote both the proliferation of possible micrometastases and the implantation of surgically disseminated tumor cells. The aim of this study is to evaluate if the preoperative subcutaneous injection of IL-2 (interleukin-2) may be able to abrogate surgery-induced immunosuppression in radically operable gastric cancer and to assess its toxicity. METHODOLOGY: This phase II study included 39 consecutive patients with histologically proven gastric adenocarcinoma (M/F 26/13; mean age 68; range 48 82) who underwent radical surgery from October 1999 to December 2000. Patients were randomized to be treated with surgery alone as controls (20 patients) or surgery plus preoperative treatment with recombinant human IL-2 (19 patients). IL 2 was administered subcutaneously, at a dose of 9,000,000 IU, for three consecutive days, followed by surgery within 36 hours from IL-2 withdrawal. We considered the total lymphocyte count and lymphocyte subset (CD4, CD4/CD8) during the preoperative period, before IL-2 administration, and on the 14th and 50th day. RESULTS: Two groups were well matched for type of surgery and extent of disease. All the patients underwent radical surgery plus D2 lymphadenectomy. At baseline, there were no significant differences in total lymphocyte and lymphocyte subsets between groups. The control group showed a significant decrease of total lymphocytes, CD4 cells, and CD4/CD8 ratio at the 14th postoperative day relative to the baseline value. Among the 22 patients evaluated in the control group 13 had a decreased of CD4 under 500 cells/mm3 (65%). Instead in the IL-2 group a significant increase was observed over the control group values of total lymphocytes and CD4 cells (14th ly total and CD4: IL-2 vs. control p<0.05). Moreover in this group only 3 patients had CD4 under 500 cells/mm3 (15%). This difference in CD4 count, is significant at the 50th postoperative day too (p=0.006). No anesthesiologic or surgical complication was seen in IL-2 treated group, with low grade of toxicity (WHO grade:1): the main effect was fever (14/19) easily manageable, with no cardiovascular complications. Furthermore, IL-2 group showed lower postoperative complications (p<0.05) and higher lymphocyte/eosinophil infiltration into the tumor (p<0.002). CONCLUSIONS: This phase II study would suggest that a preoperative immunotherapy with IL-2 is a well tolerated treatment able to prevent surgery induced lymphocytopenia. IL-2 seems to neutralize the immunosuppression induced by operation and so to stimulate the host reaction against tumor tissue (lymphocytes/eosinophils infiltration). Next randomized clinical trials could investigate the prognostic impact of IL-2 on the clinical course. PMID- 15532848 TI - Rapid urease test from non-ulcer part of stomach is superior to histology from ulcer in detection of Helicobacter pylori infection in patients with gastric ulcer. AB - BACKGROUND/AIMS: Helicobacter pylori is the major pathogenesis of peptic ulcer disease. It is important for the endoscopists to detect H. pylori infection during endoscopy. Only one endoscopic diagnostic method can be used due to the limitation of health insurance payments in Taiwan. Most endoscopists use the tissue obtained from the ulcer margins for histopathological examination and detection of H. pylori infection in patients with gastric ulcer. Whether this is suitable deserves study. METHODOLOGY: A total of 103 consecutive subjects with gastric ulcer were recruited. Biopsy specimens from the margins of gastric ulcer were sent for histological examination with modified Giemsa stain. Two biopsy specimens from the antrum and greater curvature site of mid body were embedded in rapid urease test (CLOtest). A patient was classified as H. pylori positive if either CLOtest or histology were positive. RESULTS: 57 patients had H. pylori infection. The detection rate of rapid urease test and histological examination was 96.5% (55/57) and 59.6% (35/57), respectively. Of the 3 patients who had positive histological examinations and negative urease test, only one was confirmed again to have H. pylori infection. The detection rates of rapid urease test and histological examination in different locations of ulcer (antrum/angularis/proximal stomach) were 92.6%/100%/100% and 81.5%/42.1%/36.4%, respectively. CONCLUSIONS: Our study shows that rapid urease test has higher detection rate than histological examination of the biopsy specimens obtained from the margins of gastric ulcer in diagnosis of H. pylori infection. Under the consideration of the health insurance payments limitation and elimination of false-negative detection rate of H. pylori infection, we strongly recommend the rapid urease test from the antrum and body specimens rather than from the ulcer margins for detection of the bacteria in patients with gastric ulcer disease. PMID- 15532849 TI - Nasogastric decompression after total gastrectomy. AB - BACKGROUND/AIMS: Although it is clearly known that there is no need of routine nasogastric decompression after some abdominal operations, we still do not know whether it is necessary for esophageal anastomosis. Traditionally, nasogastric decompression is mandatory after total gastrectomy complemented with esophagojejunostomy. METHODOLOGY: Consecutive 66 patients with gastric cancer who underwent total gastrectomy and esophagojejunostomy were prospectively evaluated. Patients were divided into two groups, those with nasogastric decompression and those without decompression. RESULTS: Postoperative complications were similar among the groups. Vomiting, distention, belching, hiccupping, dysphagia complaints were similar among the groups, but sore throat (100% vs. 22%, p<0.001), nausea (32% vs. 13%, p=0.054), fever (35% vs. 16%, p=0.068) and pulmonary complications (26% vs. 9%. p=0.072) were much more in the nasogastric decompression than the no-tube group. Starting oral feeding and postoperative hospital stay were similar in both groups. CONCLUSIONS: Omission of nasogastric decompression after elective total gastrectomy complemented with esophagojejunostomy did not increase postoperative complications, on the contrary it decreased postoperative fever and pulmonary problems, and improved patient comfort by decreasing sore throat and nausea. Therefore, we do not recommend the routine use of nasogastric tubes after total gastrectomy complemented with esophagojejunostomy. PMID- 15532852 TI - Facility profile: facility integrates Southern charm. Adrian & Jessie Archbold Ambulatory Care Center. PMID- 15532850 TI - Laparoscopic exclusion gastroenterostomy for palliation of gastric outlet obstruction secondary to recurrent cholangiocarcinoma. AB - The Devine exclusion gastroenterostomy is recommended as a palliative procedure for unresectable gastric carcinoma, and avoids the problem of delayed gastric emptying. It is conventionally performed by a laparotomy. We report the successful laparoscopic application of this technique in a patient with gastric outlet obstruction secondary to recurrence of a previously resected hilar cholangiocarcinoma. A 38-year-old gentleman who had undergone a left hepatectomy with caudate lobectomy, excision of extrahepatic biliary tree, D2 regional lymphadenectomy and Roux-en-Y right hepaticojejunostomy presented 6 months later with symptoms of gastric outlet obstruction. Computed tomography revealed a tumor mass in the region of the gastric antrum. Attempted endoscopic treatment with a metal stent was unsuccessful. He underwent a laparoscopic exclusion gastroenterostomy. The operative time was 200 minutes. Postoperative recovery was uncomplicated. There was no delay in gastric emptying and no recurrence of gastric outlet obstruction until the time of death 41 days later. Laparoscopic exclusion gastrojejunostomy is a feasible option for the palliation of gastric outlet obstruction caused by recurrent cholangiocarcinoma. PMID- 15532853 TI - Turbulent times: economy affects pay raises for most department heads in this year's salary survey. PMID- 15532854 TI - Winning hand: playing the odds with clinical equipment service contracts. PMID- 15532855 TI - Special delivery: management process helps rural facility build affordable central energy plant. PMID- 15532856 TI - The word on water: new CDC guidelines recommend a proactive approach to Legionella. PMID- 15532857 TI - Spot away: removing stains from health care carpeting. PMID- 15532858 TI - Bed exit alarms. AB - Bed-exit alarms alert caregivers that a patient who should not get out of bed unassisted is doing so. These alarms can help reduce the likelihood of falls and can promote speedy assistance to patients who have already fallen. But as we described in our May 2004 Guidance Article on bed-exit alarms, they don't themselves prevent falls. They are only effective if used as part of an overall fall-prevention program and with a clear understanding of their limitations. This Evaluation examines the effectiveness of 16 bed-exit alarms from seven suppliers. Our ratings focus primarily on each product's reliability in detecting bed-exit events and alerting caregivers, its ability to minimize nuisance alarms (alarms that sound even though the patient isn't leaving the bed or that sound while a caregiver is helping the patient to leave the bed), and its resistance to deliberate or inadvertent tampering. Twelve of the products use pressure-sensor activated alarms (mainly sensor pads placed on or under the mattress); three use a cord that can attach to the patient's garment, alarming if the cord is pulled loose from the control unit; and one is a position-sensitive alarm attached to a leg cuff. All the products reliably detect attempted or successful bed exits. But they vary greatly in how effectively they alert staff, minimize nuisance alarms, and resist tampering. Ease of use and battery performance also vary for many units. Of the pressure-sensor units, three are rated Preferred. Those units meet most of our criteria and have no significant disadvantages. Five of the other pressure-sensor products are Acceptable, and the remaining four are Not Recommended. All three cord-activated alarms are rated Acceptable, as is the patient-worn alarm. PMID- 15532859 TI - Bar-coded medication labeling: setting the stage for bar-code-enabled point-of care systems. AB - In February, the U.S. Food and Drug Administration (FDA) issued its final ruling requiring that all drug and biological products sold to hospitals incorporate bar codes on their labels. The ruling smooths the way for widespread adoption of bar code-enabled point-of-care (BPOC) systems, which are a valuable tool for reducing medication errors. BPOC systems help ensure that the right medications reach the right patient at the right time by allowing bar codes on a patient's ID wristband to be checked against the medication packaging. But BPOC systems will only become truly effective if medications are widely available in unit-dose packaging. Right now only about a third of all medications are available in this form. Although this situation is likely to improve, hospitals wanting to take advantage of BPOC technology soon may need to do some drug repackaging themselves (or have it done by a third party), along with a lot of other groundwork. Widespread BPOC use may still be several years away, but the time to start preparing is now. PMID- 15532860 TI - Child dies when ventilator alarms fail to warn of an occluded tracheostomy tube. PMID- 15532861 TI - Confusing restart procedure on the Puritan-Bennett Achieva ventilator could delay ventilation. PMID- 15532862 TI - Philips prism gamma camera detector can collide with the patient. PMID- 15532863 TI - Facilities' responsibilities after notification of a medical device hazard or recall. PMID- 15532864 TI - [Regarding scientific censorship]. PMID- 15532865 TI - [Affaires of the heart and gender specificity: the case for an epidemiological emergency]. PMID- 15532866 TI - [Work and prevention: who prevents and why]. PMID- 15532867 TI - [What's behind the double truth?]. PMID- 15532868 TI - [PCB contamination in Brescia: the results of the research performed by the local health unit]. PMID- 15532869 TI - [A plan for cancer control in Europe: EUROCHIP-2 (the action)]. PMID- 15532870 TI - [Road safety in Italy: epidemiology of two-wheeled motor vehicles accidents. National statistics 2000]. AB - OBJECTIVE: to describe road accidents occurred in Italy focusing, in particular, on two-wheeled motor vehicles. DESIGN: Analysis of road accidents based on current data referring to year 2000. SETTING: Italy. RESULTS: In Italy, 67,127 two-wheeled motor vehicle accidents occurred in 2000. Two crash types account for 75% of the accidents: side impact and front-side impact. Per one million kilometres travelled 0.4 cars and 1.3 two-wheeled vehicles are involved (0.7 motorcycles and 2.2 mopeds). In 2000 there were 1,229 deaths and 69,543 injured riders. Males up to forty years old are the most represented. The masculinity ratio presents a decreasing trend from motorcycles to mopeds and to cars both for killed people and for injured people. The lethality rate increases with age for all types of vehicles but for mopeds this trend is much more evident. Most of the accident occur in urban road while most of the deaths happen in extra-urban road. CONCLUSION: Mopeds and motorcycles, which are a small subset of all motor vehicles (approximately 20%), are greatly overrepresented in crashes. Considering the kilometres travelled, the risk to be involved in a crash for mopeds is estimated to be 32.6 times higher than the comparable risk for cars and for motorcycles it is 17 times higher. The risk of death for riders is two times the risk for cars. PMID- 15532871 TI - [Geographical distribution of mortality from malignant pleural neoplasms and of former asbestos-exposed workers in the Campania Region]. AB - AIM: The purpose of the present paper is to describe the geographical distribution at municipality level of mortality from malignant pleural neoplasms in the Campania Region, along with the distribution of former asbestos-exposed workers. A GIS (Geographical Information System) application has been developed to integrate exposure and mortality data. STUDY DESIGN: The number of asbestos workers by municipality has been estimated by merging data provided by the National Institute for Insurance against Occupational Accidents (INAIL), the Ministry of Labour and an association of formerly exposed workers (AUSER Flegrea). The number of deaths from malignant pleural neoplasms by municipality has been derived from published studies regarding two consecutive periods: 1988 1994 and 1995-1997. RESULTS: In the first period most deaths occurred in areas with a high number of formerly exposed asbestos workers, whilst in the second period mesothelioma mortality increased extending to other areas without registered exposed workers. CONCLUSION: This finding may be attributable to the presence of atypical occupational exposures, not reported by the current information systems, or to environmental, non occupational exposure patterns. PMID- 15532872 TI - [Health Search--Research Institute of the Italian Society of General Practice: the creation of a research database in general practice]. AB - OBJECTIVE: To describe a database used in general practice by analysing the characteristics of the physicians providing the data and the completeness of the data recording on the basis of the indicators of interest (smoking habits, weight, height, and the prevalence of hypertension). DESIGN: Descriptive study and multiple linear regression analysis of the relationships between structural variables and outcomes. SETTING: General practitioners (GPs) belonging to the Italian Society of General Medicine (SIMG), and enrolled in Health Search (HS). POPULATION: Six hundred and ninety-five voluntarily enrolled GPs and their patients. PRINCIPAL OUTCOMES: Descriptive statistics (mean and median values, standard deviation, frequencies) concerning the general characteristics of the GPs, the ways in which they connect to the network and their use of the clinical data management software, and epidemiological measures concerning the prevalence of hypertension among their patients, and the recording of weight, height and tobacco smoking habits. RESULTS: The geographic distribution of the GPs is homogeneous: 317 in Northern Italy (12.52 physicians per 10(6) patients), 134 in Central Italy (12.30 physicians per 10(6) patients) and 244 in Southern Italy and the Islands (11.89 physicians per 10(6) patients). Five hundred and ninety-five (85.6%) are males; their mean age is 46.7 years (SD +/- 3.8); the mean number of patients per physician is 1128; 69% have at least one post-graduate specialisation; 49% practise alone; and they work for a mean of 30 hours per week (SD +/- 13). Five hundred and fifty-three GPs were actually connected to the network as of January 2003. The data relating to the years 2000-2001 show an increase in the number of connected physicians (34 in 2000 and 261 in 2001) and in the number of connections (a median of 12 in 2000 and 17 in 2001). The GPs contact more than 80% of their patients every year. The frequency of the recording of data concerning smoking habits, weight and height increases in proportion with the frequency of connections. The prevalence of hypertension among the patients included in the survey is 5.4%. Multiple regression analysis showed that the variability in the prevalence of hypertension was not influenced by the frequency of PC use but by the different structure of the age classes of the patients. CONCLUSIONS: This study of the variables relating to GPs, their clinical practices, and their use of computerised records (examined by means of the chosen indicators) shows that the regular and complete recording of the principal data of health interest is feasible insofar as it is not biased by the characteristics of the GPs, and advantageous. The construction of the database therefore represents a first step towards the initiation of routine research into general practice in Italy. PMID- 15532873 TI - [HIVAb, HCVAb and HBsAg seroprevalence among inmates of the prison of Bologna and the effect of counselling on the compliance of proposed tests]. AB - OBJECTIVES: The aims of the study were to evaluate the HIVAb, HCVAb and HBsAg seroprevalence among Italian and foreign inmates of the prison of Bologna, to evaluate if the extensive counselling of "new" inmates has significantly enhanced adherence to laboratory tests. DESIGN: The serological status was determined by a blood withdraw following the informed consent. Before asking their consent, patients were informed by cultural mediators who had been instructed about the aims of the study/exam during introductory meetings. The initial step managed by mediators was followed by further individual counselling interventions, carried out by hospital infective disease unit, prison and prison drug abuse service physicians. The laboratory tests were performed in an external structure. SETTING: Prison of Bologna. PARTICIPANTS: The study was conducted on 433 subjects among a whole population of 900 inmates in the local prison: 390 subjects were males (90.1%) and 43 were females (9.9%). The median age of the whole population was 34.86 years (+/- 9.9). The studied population counted 147 (33.9%) intravenous drug users (IDU) and 286 not addicts (66.1%). As regards nationality, 212 subjects were Italian (48.9%) and 221 (51.1%) foreigners. Among the total 433 inmates considered, 78 (18%) were known as previous IDU with conviction history or condemned to long term sentences, while 59 (13.6%) were inmates recently convicted active IDU assisted by the internal drug abusers service. The third group was composed by 296 inmates imprisoned during the summer (103 Italians and 193 foreigners) self declared not IDU. RESULTS: A. 12.5% of inmates were HIV positive, 8.1% HBV positive and 31.1% HCV positive. 25 subjects were found positive both to HIV and HCV; 1 both to HIV and HBV and 5 to HIV, HBV and HCV. HIV positivity is more common among Italian vs. foreigners inmates, among IDU vs. not IDU. HCV positivity is more common among Italian vs. foreigners inmates, and among IDU vs. not IDU. The distribution of HBV seropositivity among the different groups shows no statistically significant differences. All subjects receiving multi-focal counselling reached better compliance levels: 10% vs. 1% for HIV, 16% vs. 1% for HBV and 35% vs. 0.3% for HCV, with statistically significant coefficients of contingency. CONCLUSIONS: In the prison of Bologna drug addiction is prevalent in italian seropositive personers and it is often associated with HIV and HCV positivity. Foreign inmates, mostly in the not-IDU group, show a lower prevalence of these two infections. Multi-focal counselling before test increased significantly the adherence of inmates to the study. PMID- 15532874 TI - [A series of cases of sinonasal cancer among shoemakers and tannery workers]. AB - A series of cases of sinunasal cancer has been observed among shoe-makers (11 cases) and tannery workers (8 cases) in the leather production area in Tuscany in the period 1990-2002. We stress the possible etiologic role of tannins both in leather and in wood industry. PMID- 15532876 TI - [Prevention of doping in sports: epidemiological issues and ethical implications]. AB - Owing to a widespread diffusion, the consumption of banned and potentially harmful substances in sports has become a problem for the public health. Current estimations of the prevalence of doping in sports are rather uncertain, as most investigative tools do not reflect an absolute statistical power. However, the emerging scenario reflects a concerning underestimation by Structures and Institutions that should establish definitive rules and set reliable controls. Owing to restricted resources, prevention and fight against doping must be supported by meditated and rational strategies, with the aim to identify suitable contests and accurate procedures, considering carefully ethical issues that may arise from the positivity of the athletes to antidoping controls. PMID- 15532875 TI - [Life expectancy as an indicator of outcome]. AB - Usually survival studies evaluate the percentage of subjects alive or disease free at a given point in time (cumulative survival percentage). These studies require a lengthy period both for follow up and for the collection of an exhaustive number of cases. In addition, for cancer types with a sharp gradient of cumulative mortality, the estimate may be unstable. On the database of children with Acute Lymphoblastic Leukaemia (ALL) collected by the Childhood Cancer Registry of Piedmont (CCRP) in 1979-1991, we have estimated an alternative time-dimensional index to estimate both patients' life expectancy of patient and number of years gained to death. These estimates have been compared in subsets of the database corresponding to different periods of diagnosis and efficacy of treatment. Life expectancy has been evaluated as the area under the survival curve, on the assumption that after a number of years since diagnosis, survivors have the same mortality rates of the general population. Clinicians are invited to take into consideration the approach described in the present note. PMID- 15532877 TI - [Practice guidelines, CME and epidemiologic research in general practice: a consistent triad?]. AB - A multidisciplinary effort involving epidemiologists, general practitioners and specialists is critical for finding valid and feasible answers, by asking the right questions through observational and experimental studies. Clinical practice guidelines may represent a good starting point of this research process, highlighting which information needs exist in everyday practice; logically, guidelines also represent an ideal end, as a means of transferring any useful knowledge produced into practice. In between, this research process may have an added educational value and represent an active and valuable means of achieving continuing medical education credits. Consistency of the research-education guidelines triad is discussed in this paper by describing methods and results of a practical research experience--a cohort study for evaluating prevalence, incidence, chronicity and management of digestive disorders--involving epidemiologists, gastroenterologists and thirty-five general practitioners of the Emilia-Romagna region. PMID- 15532878 TI - [Female genital mutilation and substitutive ritual: comment from the point of view of cultural anthropology]. PMID- 15532879 TI - The importance of references to a peer reviewed article. PMID- 15532880 TI - An ethicist's commentary on good versus natural death. PMID- 15532881 TI - The first Canadian indigenous case of bovine spongiform encephalopathy (BSE) has molecular characteristics for prion protein that are similar to those of BSE in the United Kingdom but differ from those of chronic wasting disease in captive elk and deer. AB - Brain tissue from a case of bovine spongiform encephalopathy (BSE) from Alberta was subjected to a Western immunoblotting technique to ascertain the molecular profile of any disease-specific, abnormal prion protein, that is, prion protein that is protease-resistant (PrP(res)). This technique can discriminate between isolates from BSE, ovine scrapie, and sheep experimentally infected with BSE. Isolates of brain tissue from the BSE case in Alberta, 3 farmed elk with chronic wasting disease (CWD) from different parts of Saskatchewan, and 1 farmed white tailed deer with CWD from Edmonton, Alberta, were examined alongside isolates of brain tissue from BSE, ovine scrapie, and sheep experimentally infected with BSE from the United Kingdom (UK). The molecular weights of PrP(res) and the cross reactions to 2 specific monoclonal antibodies (mAbs) were determined for each sample. The BSE isolates from Canada and the UK had very similar PrP(res) molecular weights and reacted with only 1 of the 2 mAbs. The PrP(res) isolated from both elk and white-tailed deer with CWD had a higher molecular weight profile than did the corresponding PrP(res) from the scrapie and BSE isolates. The PrP(res) from CWD cases cross reacted with both mAbs, a property shared with PrP(res) in isolates from scrapie but not with PrP(res) isolates from BSE or sheep experimentally infected with BSE. The results from this study seem to confirm that the PrP(res) isolated from the BSE case in Alberta has similar molecular properties to the PrP(res) isolated from a BSE case in the UK, and that it differs in its molecular and immunological characteristics from the CWD and scrapie cases studied. PMID- 15532883 TI - Dissolution of infection-induced struvite bladder stones by using a noncalculolytic diet and antibiotic therapy. AB - An 8-year-old, female spayed miniature schnauzer was presented for pollakiuria and gross hematuria. Infection-induced struvite urolithiasis with concurrent bacterial urinary tract infection was diagnosed. The treatment is described, followed by a brief discussion of struvite stones and their medical management. PMID- 15532884 TI - Vitamin D-responsive rickets in neonatal lambs. AB - An unusual clinical presentation of rickets in nursing lambs is described. Two sets of twin nursing lambs were presented for enlarged joints and mild angular deformity. Rickets was suspected, based on radiographic evidence. Vitamin D deficiency was determined to be the cause, based on serum concentration and response to therapy. PMID- 15532882 TI - Evaluation of oxidative stress markers for the early diagnosis of allograft rejection in feline renal allotransplant recipients with normal renal function. AB - The purpose of this study was to identify oxidative damage to renal allografts during graft rejection by evaluating changes in oxidative markers and plasma lactate levels in feline renal allotransplant recipients. Heterotopic renal allotransplantations were performed between 8 adult feline cross-matched donors. Following 14 d of immunosuppression, the drugs were discontinued to allow allograft rejection. Baseline and serial postoperative evaluations of serum creatinine, plasma lactate, plasma thiobarbituate reactive substances (TBARS), plasma creatol, urine creatol, and renal sonographic cross-sectional area were performed. When sonographic evaluation revealed the absence of blood flow to the allograft, the rejected kidney was nephrectomized and evaluated histopathologically. Allograft rejection occurred in all cats by day 26. A significant elevation in body temperature occurred during the rejection period. No significant change was observed between any of the time periods for plasma TBARS, creatol, or urine creatol. There was a significant decrease in plasma lactate levels throughout the study. Markers of oxidative stress from venous blood did not reflect renal allograft rejection in cats with a normally functioning native kidney. Renal allograft rejection may be associated with significant increases in body temperature and warrants further investigation. PMID- 15532885 TI - Bovine familial convulsions and ataxia in Saskatchewan and Alberta. AB - Bovine familial convulsion and ataxia, a heritable disorder, is present in crossbred Aberdeen Angus cattle in western Canada. Though the prevalence appears to be low, this genetic defect might become widespread if not controlled. Characteristic histological lesions were seen in the lingula, uvula, or both, of all affected calves regardless of their age. PMID- 15532886 TI - Phosphate enema toxicosis in a pygmy goat wether. AB - Phosphate enema toxicity was diagnosed in a 7-month-old, castrated male, pygmy goat. On presentation, clinical findings included mild depression, tachycardia, tachypnea, rumen stasis, muscle tremors, hypocalcemia, hypokalemia, hypochloremia, hyperphosphatemia, azotemia, and metabolic acidosis. Fluid diuresis and parenteral antimicrobial therapy resulted in recovery after 3 d of treatment. PMID- 15532887 TI - Bovine cysticercosis: preliminary observations on the immunohistochemical detection of Taenia saginata antigens in lymph nodes of an experimentally infected calf. AB - A newly developed immunohistochemical test was used for the first time to demonstrate the presence of Taenia saginata (Cysticercus bovis) antigens in the lymph nodes of a heifer calf experimentally inoculated with Taenia saginata eggs. The new test should aid in the differential diagnosis of eosinophilic lymphadenitis in cattle. PMID- 15532888 TI - A system response to an outbreak of enzootic pneumonia in grow/finish pigs. AB - A Mycoplasma hyopneumoniae-negative commercial swine production system broke with enzootic pneumonia at their grow/finish site in southern Manitoba in October, 2003. System responses included feed medication, depopulation, delayed shipment of pigs to the infected site, vaccination of at risk sow herds, and disinfection when grow/finish site depopulation was completed. PMID- 15532889 TI - Unilateral renal cell carcinoma in a Labrador retriever. AB - A 7-year-old, neutered male, Labrador retriever was presented with a history of hematuria. Radiographic and ultrasonographic studies led to a presumptive diagnosis of unilateral renal neoplasia. Unilateral nephroureterectomy was performed. Histopathologic examination of the resected mass confirmed the diagnosis of renal cell carcinoma. Six weeks postoperatively, the dog was well. PMID- 15532890 TI - Renal lipomatosis in a lamb. PMID- 15532891 TI - What's new in orthopedics? Part 1. PMID- 15532892 TI - The art of referring: to whom to refer. PMID- 15532893 TI - Diagnostic dermatology. PMID- 15532894 TI - [Imatinib--a new perspective in the treatment of tumors]. AB - Among novel promising approaches to anticancer therapy belongs the targeting inhibition of signal transduction. This review outlines present-day experiences with imatinib (Glivec), a potent inhibitor of the tyrosine kinases bcr-abl, c-kit and platelet-derived growth factor receptor kinase. Due to inhibition of bcr-abl tyroxine kinase, imatinib has rapidly become the standard therapy for chronic myelocytic leukemia; inhibition of c-kit receptor explains its effectivity in the treatment of patients with gastrointestinal stromal tumors. Another known target of imatinib is tyrosine kinase of PDGFR, which is activated in numerous malignancies, particularly in dermatofibrosarcoma protuberans. Discovery of the novel fusion gene in hypereosinophilic syndrome (FIPILI-PFGFRA, whose product is an imatinib sensitive protein kinase) permitted to treat successfully this event. Possible combination of imatinib with conventional chemotherapeutic drugs and other key signal transduction inhibitors are mentioned. PMID- 15532895 TI - [Acute HIV infection]. AB - Acute HIV infection occurs in 85% of people recently infected with HIV-1 after an incubation period of 2-6 weeks. Diagnosing the acute HIV infection is difficult because the symptoms are non-characteristic and include fever, pharyngitis, and skin rash. Early recognition of HIV infection is, of course, maximally desirable in order to prevent further transmission of the infection and because the early treatment can slow down later progression of the disease. PMID- 15532896 TI - [Conservative treatment for the penis carcinoma]. AB - No explicit recommendation has been determined in a treatment for the verified squamous cell penis carcinoma till now. The application of ionizing radiation is included in traditional treatment methods for this disease, in addition to surgical operations and chemotherapy cure. It is possible to apply external radiotherapy or brachy-radiotherapy (a moulage or an intersticial application) as well as their combination. In individual cases it is possible to use chemo radiotherapy. In the case of the localized tumor a curative radiotherapy can be used as it is more save towards this organ. Radiotherapy has its place in the neoadjuvans treatment with the goal to reduce the disease extent. It can be also used as an adjuvans--postoperatively and paliatively. This review is describing principles of the radiotherapy treatment for this disease. PMID- 15532897 TI - [Tuberculosis in the Czech Republic in 2003]. AB - BACKGROUND: 1162 new cases of tuberculosis were diagnosed in the Czech Republic in 2003; pulmonary tuberculosis represented 942 cases and in 660 cases the diagnosis of tuberculosis was bacteriologically confirmed. METHODS AND RESULTS: 355 cases were smear positive tuberculosis. The increase of certain TB cases (70%) in 2003 compared to 2002 is not statistically significant (p = 0.031). The notification rate (incidence) in 2003 was 11,4/100 000 all cases of tuberculosis, 9,2/100 000 pulmonary tuberculosis and 6,5/100 000 certain cases of tuberculosis. CONCLUSIONS: Compared to 2002 a decline of all forms of tuberculosis was observed, this decline however was not statistically significant. The increase in bacteriologically confirmed tuberculosis and smear positive tuberculosis was also not statistically significant. PMID- 15532898 TI - [Results of treatment of malignant stenoses of the respiratory tract in a long term study]. AB - BACKGROUND: :The malignant stenoses complicate the course of the disease in about 50% of patients with lung tumors. Management of them can influence the quality of life of the afflicted. There is no general agreement about optimal methods of their therapy, of selection of the best methods and also there is only few data about survival of these patients. METHODS AND RESULTS: 130 patients with tumors stenoses of the airways were prospectively followed in the 1998-2003 period, 93 of them suffered from lung cancer. Besides other treatment of the tumor they underwent also interventional bronchoscopy (Nd YAG laser, electrocautery i.e.). The airways were opened completely in 62% of the set of patients. The interventional bronchoscopy was complicated in 9% of patients, 32 patients lived at the 31st December of 2003, 98 of them had died. One year survival was 23.1%, median survival of the whole set of patients was 7 months. The patients with lung cancer had significantly worse survival than patients with other types of lung tumors, 6 months versus 10 months. There were no significant survival differences of the whole set according to the achieved grade of recanalisation, or according the inclusion of the brachytherapy to the set of interventional methods. The patients with inserted stent had significantly worse prognosis than the those without stenting. To homogenize the study group, the subset of patients with epidermoid lung cancer in the stage IIIB were selected (n = 51). Significantly longer survival of patients with complete recanalisation, insignificant longer survival in patients who underwent also brachytherapy and significantly shorter survival in patients with stenting were proved in this subgroup of patients. CONCLUSIONS: The interventional bronchoscopy has a low rate of complications and in most cases it has a palliative effect. The survival of patients is more influenced by the extension and the type of the tumor, their general status, polymorbidity, age etc. than by effect or selection of interventional bronchoscopical methods. PMID- 15532899 TI - [Dispersion of QT intervals--a myth or a diagnostic symptom?]. AB - BACKGROUND: QT interval dispersion (QTd) is conventionally interpreted as a result of repolarization heterogeneity in ventricular myocardium. However, another concept of QTd origin has been discussed recently, suggesting that different projections of the repolarization vector into individual ECG leads could be responsible for the differences in QT interval duration. Moreover, the reproducibility could be influenced by factors both electrocardiographic (T wave amplitude, U wave) and extracardiac (noise, ECG measures). In the presented study we have followed the QTd in two groups of patients with proved changes of an electric heart field. METHODS AND RESULTS: Studied groups: 1. Control group, 2. Healthy pregnant women, 3. Patients treated with dosulepine. QT interval was measured from 80 unipolar chest leads used for body surface potential mapping. The QTd was significantly higher in both experimental groups in comparison with the control group (p < 0.001). Significant correlation was found between the QTd and dosulepine plasma level (p < 0.001). Also amplitude of the T wave loop was in both groups decreased and its width increased (both p < 0.001). CONCLUSIONS: If appropriate procedure of measurement is used, the QTd is significantly increased in many physiological and pathological states. Clinical relevancy of borderline increased values has to be interpreted very carefully. PMID- 15532900 TI - [Significance of the mathematical model of cardiac electrical field for the interpretation of experimental data]. AB - BACKGROUND: In concurrence of our recent findings of the elevation of QT dispersion (QTd) in the group of pregnant women, mathematical approaches were developed aimed to give possible geometrical explanation whether the observed changes result from the rotation or from the changed position of the heart. METHODS AND RESULTS: Mathematical model of the cardiac electrical field approximated as a time variable dipole in a homogenous spatial conductor was developed. From the experimental vectocardiographic records, representing time course of the cardiac dipole, body surface potential maps were calculated on the basis of the model. To validate the adequacy of the model, the reconstructed electrocardiograms were compared with the empiric data. To determine the effects of rotation, original empiric VCG data of the control group were transformed accordingly the hypothetic pregnancy related changes. Calculated surface electrocardiograms were then compared with empiric cardiograms of the pregnant women. CONCLUSIONS: Based on the results, several conclusions can be drawn: 1) QT dispersion is associated also with the geometrical relations between the direction of cardiac vector during the terminal phase of repolarization and the direction of axes in the given system of leads. The dispersion then has its typical occurrence at the thoracic surface--minimums of the QT duration are found in the plane perpendicular to the axis of the terminal vector lead. 2) When the duration of repolarization is estimated from the classic thoracic leads within the phisiological variations of terminal-depolarization vector orientations, can exist that in some cases the minimum of QT interval is and in others it is not recorded by the lead system. Value of QT dispersion between these two extremes will be significantly different. 3) In case of the horizontal declination of the heart, the ECG signal in most of the leads of the body surface mapping has a higher voltage than in case of vertical declination due to a smaller angle between axes of the terminal vector and most of the leads. Such fact will contribute to more accurate reading of the T wave end and to the estimation of QT interval, usually with smaller value of QTd. 4) The change of the cardiac electrical field corresponding to the changed position of the heart (rotation) does not result by itself in QTd changes, if it is evaluated from the records from the whole thorax. Obversely, horizontalization of the heart contributes more to the evaluation of lower QTd values, as it is given above. 5) More then the result of geometrical changes, QT dispersion found in the group women in high level of pregnancy is an effect of changes in the T loop morphology, which was observed in this group. Another possible explanation of the observed dispersion is the non-dipolar character of the electrical field changes during pregnancy. PMID- 15532901 TI - [Results of treatment of esophageal atresia]. AB - BACKGROUND: The aim of the study was to analyze morality, incidence of complications and influence of various prognostic factors on the outcome of patients with esophageal atresia. A retrospective analysis of the records of 109 consecutive cases of esophageal atresia and/or tracheoesophageal fistula operated in 1992-2001 was undertaken. Primary anastomosis was achieved in 68 (62.4%) patients, in 7 (6.4%) procedures aimed at esophageal substitution in future were performed and in 34 (31.2%) delayed anastomosis was primarily planned. METHODS AND RESULTS: Major associated anomalies were found in 55 (50.4%), cardiac anomalies in 19 (17.4%), and more than one associates anomaly in 16 (14.7%) patients. The most frequent surgical complication was anastomotic leak in 11.9%, septic complications were the most frequent and occurred in 66 (60.5%) patients. Gastroesophageal reflux was diagnosed in 30.3% and treated surgically in 14.7% of infants. The overall mortality was 20.2% (22 patients). The most frequent cause of death was sepsis and/or pneumonia. Long gap esophageal atresia was not found to be a significant factor in mortality. Factors significantly increasing mortality were birth weight below 2000g (OR 3.94), cardiac anomalies (OR 3.95), and mechanical ventilation before the operation (OR 2.97). CONCLUSIONS: Analysis of the outcome suggests that improved strategy in treating infants with multiple anomalies and more frequent attempts at primary anastomosis even in children in sub optimal general condition at diagnosis could decrease mortality. PMID- 15532902 TI - [Radical surgery in the complex treatment of hepatoblastoma]. AB - BACKGROUND: Evaluation of treatment results in children with hepatoblastoma--a retrospective study of clinical and pathological data of surgically treated children. METHODS AND RESULTS: At the clinic of authors, 28 children were operated for hepatoblastoma during 1991 to 2002. Average age of patients was 2.2 years. When tumors were classified according to PRETEX system, 3 children were in the stadium I, 12 children in stadium II, 12 children in stadium III, and one child in the stadium IV. At the diagnosis, lung metastases were found in 3 children. Till 1996 the treatment was initiated by a primary operation, since 1996 by a preoperative chemotherapy. Chemotherapy was administered according to the SIOP protocols. Primary operation was done in 13 children, 15 children were operated after the pre-operation chemotherapy. Liver resection included right sided lobectomy in 7 cases, extended right-sided lobectomy in 4 cases, left-sided lobectomy in 8 cases, right-sided trisegmentectomy in 5 cases, left-sided trisegmentectomy in 3 cases. In our studied group the three years of event free survival was achieved in 75%, overall survival in 86%. Four children died, two of them because of the progression of the disease, two for the complication during the therapy. CONCLUSIONS: Combination of the radical surgical resection with preoperative and postoperative chemotherapy enables successful treatment of the childhood hepatoblastoma. PMID- 15532904 TI - [Type I diabetes mellitus and associated autoimmune diseases in the first-degree relatives of diabetic children: questionnaire based study]. AB - BACKGROUND: Type 1 diabetes mellitus (T1DM) is associated with increased incidence of other autoimmune diseases. The shared genetic background may play a role in the disease pathogenesis. The aim of our study was to assess the prevalence of T1DM and other autoimmune disorders in the first-degree relatives of diabetic children. METHODS AND RESULTS: Data were retrospectively obtained using structured questionnaires from 868 diabetic children younger than 18 years (434 girls and 434 boys, age 12.5 +/- 4.0, mean +/- SD) and their 2704 relatives. The control group included 1466 non-diabetic schoolmates and friends (796 girls, 670 boys, age 11.9 +/- 4.5) and their 4510 first-degree relatives. In the questionnaire we asked about occurrence of thyroid and celiac disease in cases and controls, and about occurrence of T1DM, thyroid and celiac disease in their first-degree relatives. We observed significantly higher prevalence of T1DM in fathers (4.4% vs. 0.8%), mothers (2.0% vs. 0.5%) and siblings (2.5% vs. 0%) of diabetic children compared to controls. Thyroid disease was found significantly more in diabetic children (10.0% vs. 1.9%) and their siblings (3.1% vs. 1.7%). Prevalence of celiac disease was also higher in diabetic children than in controls (3.2% vs. 0.5%), but it does not differ in their first-degree relatives. CONCLUSIONS: We found significantly higher prevalence of thyroid and celiac disease in T1DM children than in controls. Targeted screening and early detection of thyroid and celiac diseases in T1DM patients are likely to be necessary. We observed an increased prevalence of T1DM and thyroid disease in first-degree relatives of diabetic children, however screening of autoimmune diseases associated with T1DM in the first-degree relatives remain controversial. PMID- 15532903 TI - [Treatment of congenital diaphragmatic hernia in neonates]. AB - BACKGROUND: Treatment of the Congenital Diaphragmatic Hernia in neonates with early manifestations has been accompanied with significant mortality. Authors present the treatment protocol employed in such children and results from the period January 1994 to December 2003. METHODS AND RESULTS: Since January 1994 to December 2003 96 children were cured for the Congenital Diaphragmatic Hernia. Ratio F:M was 36:60. 29 children (30%) were diagnosed prenatally, the remaining newborns were diagnosed on the basis of thoracic X-ray, which was required for the early signs of respiratory insufficiency. The postnatal treatment protocol included circulation-ventilation stabilisation, operation, early extubation and initiation of the peroral nourishment. Altogether 78 (81%) of children were operated after the stabilization using the standard laparotomy method, 5 children (6%) were operated with extra corporal membrane oxygenation, in 20 children (26%) a Goretex patch was used to cover an extensi e defect in the diaphragm. Overall survival was 74%, survival of operated children was 91%, 75% of children with a patch survived, survival of children operated with ECMO was 40%. Relapse occurred in one child (1%) with a patch. CONCLUSIONS: Specification of the prenatal diagnosis and collection of patients with CDH into tertiary centres with ECMO possibility can improve prognosis of neonates with the malformation. PMID- 15532905 TI - [Mobile thrombus at the origin of the internal carotid artery]. AB - Stroke is the third leading cause of death in many countries worldwide. The vast majority of acute strokes is ischaemic, caused often by embolisation from the large arteries of the head and neck, less frequently by cardiogenic embolisation. A case report of a patient with mobile thrombus at the origin of the internal carotid artery diagnosed by duplex sonography is presented. The patient refused surgery, two days later a recurrent severe stroke occurred caused by embolisation of the found thrombus. PMID- 15532907 TI - [Medical and philosophical discussion of human nature]. PMID- 15532906 TI - [Correlation of 99mTc-dTPA renal scintigraphy with ACE inhibitor and ultrasound findings in screening for renal artery stenosis]. AB - BACKGROUND: ACE inhibitor renal scintigraphy is widely accepted as a useful test in screening of unilateral renal artery stenosis. Another screening examination is ultrasonography, which detects haemodynamic changes with evaluation of resistive index (RI). The aim of the present study was correlation of 99mTc-DTPA captopril renography and duplex ultrasound findings in diagnosis of renal artery stenosis before angiography, that is used as "gold standard". Angiography was performed in 35 patients and their findings were correlated with scintigraphy and ultrasonography. METHODS AND RESULTS: 93 patients (49 male and 44 female) aged 27 to 84 years were examined. Ultrasonography was performed in all patients with evaluation of RI. Estimation of scintigraphy was performed in accordance with interpretative criteria of 99mTc-DTPA captopril renography, which were established at the 9th Symposium of radionuclides in nefro-urology (Santa Fe 1995) and Guideline for diagnosis of renovascular hypertension (1998). 1) Agreement positive and negative findings on scintigraphy and ultrasonography were in 61/93 patients (66%). 2) Within the group of 35 patients with angiography, the correlation between scintigraphy and ultrasonography findings was found in 27 patients (77%). 3) When scintigraphy and angiography were correlated, the correctly diagnosed renal artery stenosis was found in 27 patients with sensitivity (77%). 4) When sonography and angiography were correlated, the correctly sonographicaly diagnosed renal artery stenosis was found in 26 patients with sensitivity (74%). CONCLUSIONS: On the basis of our results and according to our experience with both examinations we can conclude, that screening of unilateral renal artery stenosis with using these nonivasive methods can help to select better the indicated patients for angiography. PMID- 15532909 TI - Editorial transition: ave atque vale. PMID- 15532910 TI - Hospital report cards: intent, impact, and illusion. AB - The public is currently being bombarded with a growing number of hospital report cards revealing hospital-specific outcome data. The original intent of these report cards was to provide information to the public to enable them to make a more educated choice in selecting health care services and to stimulate provider activities to improve outcomes of care. Serious issues have been raised about the accuracy, validity, meaningfulness, and interpretability of these reports, which has affected both provider and consumer response. With public image and financial implications on the line, hospitals need to be aware of these reports and take the necessary steps to improve outcomes of care. PMID- 15532911 TI - Improving health care experiences of persons who are blind or have low vision: suggestions from focus groups. AB - Persons who are blind or have low vision face special challenges in obtaining care that is safe, effective, timely, and patient centered. To explore perceptions of care and recommendations for improvements, we conducted 8 interviews with experts and 2 focus groups with 19 persons, all of whom are blind or have low vision. Interviewees perceived that they confront special barriers to care because of being blind or having low vision. Barriers fell into 4 broad categories: basic respect, including concerns about physicians thinking they cannot participate fully in their own care; communication barriers, including difficulties interacting with physicians and office staff; physical access barriers, including difficulties getting to and around physicians' offices; and information barriers, including receiving written materials in inaccessible formats (eg, not in Braille, large print, or audiotape). Using common courtesy and individualized communication techniques, physicians and office staff could improve health care experiences of blind and low-vision patients. PMID- 15532912 TI - Identifying hypertension-related comorbidities from administrative data: what's the optimal approach? AB - The objective was to determine the best strategy for identifying outpatients with hypertension-related diagnoses using Veterans Affairs (VA) administrative databases. We reviewed 1176 outpatient charts from 10 VA sites in 1999, taking the presence of 11 diagnoses relevant to hypertension management as the "gold standard" for identifying the comorbidity. We calculated agreement, sensitivity, and specificity for the chart versus several administrative data-based algorithms. Using 1999 data and requiring 1 administrative diagnosis, observed agreement ranged from 0.98 (atrial fibrillation) to 0.85 (hyperlipidemia), and kappas were generally high. Sensitivity varied from 38% (tobacco use) to 97% (diabetes); specificity exceeded 91% for 10 of 11 diagnoses. Requiring 2 years of data and 2 diagnoses improved most measures, with minimal sensitivity decrease. Agreement between the database and charts was good. Administrative data varied in its ability to identify all patients with a given diagnosis but identified accurately those without. The best strategy for case-finding required 2 diagnoses in a 2-year period. PMID- 15532913 TI - The effect of physician feedback and an action checklist on diabetes care measures. AB - The objective was to evaluate whether physician feedback accompanied by an action checklist improved diabetes care process measures. Eighty-three physicians in an academic general medicine clinic were provided a single feedback report on the most recent date and result of diabetes care measures (glycosylated hemoglobin [A1c], urine microalbumin, serum creatinine, lipid levels, retinal examination) as well as recent diabetes medication refills with calculated dosing and adherence on 789 patients. An educational session regarding the feedback and adherence information was provided. The physicians were asked to complete a checklist accompanying the feedback on each of their patients, indicating requested actions with respect to follow-up, testing, and counseling. The physicians completed 82% of patient checklists, requesting actions consistent with patient needs on the basis of the feedback. Of the physicians, 93% felt the patient information and intervention format to be useful. The odds of urine microalbumin testing, serum creatinine, lipid profile, A1c, and retinal examination increased in the 6 months after the feedback. The increase was sustained at 1 year only for microalbumin and retinal exams. There was no significant change in refill adherence for the group overall after the feedback, although adherence did improve among patients of physicians attending the educational session. No significant change was noted in lipid or A1c levels during the study period. In conclusion, a simple physician feedback tool with action checklist can be both helpful and popular for improving rates of diabetes care guideline adherence. More complex interventions are likely required to improve diabetes outcomes. PMID- 15532914 TI - Measuring a hospital's ability to improve. AB - The aim of this study was to test whether a recently developed measure of Continuous Quality Improvement (CQI) implementation can provide health care researchers and administrators with a tool to assist in understanding and with developing an appropriate structure for improvement efforts in hospitals. Two hundred respondents from 40 Missouri hospitals completed a 28-item survey addressing 8 domains of CQI. Overall, hospital scores showed low implementation of a structure that supports improvement efforts. All survey domains showed acceptable psychometric results. Leadership proved to be the most important domain of CQI because it differentiated well between all levels of the scale. Because of its ease of administration and analysis, and its reliability, validity, and level differentiation results, the researchers recommend the widespread use of this tool to understand and develop a hospital's organizational structure to support improvement activities. PMID- 15532915 TI - Development of an internal dynamic web site to promote quality assurance in a clinical laboratory. AB - In clinical laboratories, one challenging quality assurance objective is to maintain standardized practices. Meeting this objective entails ensuring information flow, which is necessary to smooth running of the laboratory. To facilitate information flow, we developed an internal quality Web site on our local network. The dynamic generated pages of the site were constructed with EasyPHP v.1.6, a complete freeware package providing PHP dynamic language and databases. The site comprises various sections: general news, specific laboratory units news, documents (quality manual, guidelines, emergency processes), schedules, National Quality Control results, forum, etc. Five to 10 pages are updated each week. This work was facilitated by the use of PHP-written pages and data tables, which enable us to record in real time the operation of our assurance quality project and to improve traceability. This approach could be extended to other aspects of quality management and could help meet the future IS015189 standard requirements. PMID- 15532916 TI - Presidential address given at the 70th Annual Meeting of the American Mosquito Control Association and the 29th Annual Meeting of the Mid-Atlantic Mosquito Control Association, february 2004. PMID- 15532917 TI - Culex (Culex) declarator, a mosquito species new to Florida. AB - One specimen of a mosquito new to Florida, Culex declarator, was first found in 1998 in Indian River County. A 2nd specimen was collected in 2002. Beginning in September 2003, Cx. declarator adults were regularly encountered in routine mosquito surveillance sampling, with more than 300 specimens appearing in 45 collections. Prior to our find, the U.S. distrubution was thought to be restricted to south Texas. The full extent of this species' distribution in Florida has yet to be determined. PMID- 15532918 TI - A checklist of mosquitoes (Diptera: Culicidae) of Pondicherry, India with notes on new area records. AB - A checklist of mosquito species for Pondicherry, India, is presented based on collections made from November 1995 to September 1997. Mosquitoes of 64 species were found belonging to 23 subgenera and 14 genera, Aedeomyia, Aedes, Anopheles, Armigeres, Coquillettidia, Culex, Ficalbia, Malaya, Mansonia, Mimomyia, Ochlerotatus, Toxorhynchites, Uranotaenia, and Verrallina. We report 25 new species for Pondicherry. PMID- 15532919 TI - Commercial mosquito trap and gravid trap oviposition media evaluation, Atlanta, Georgia. AB - Field trials evaluating the effectiveness of selected gravid trap oviposition media and commercially available mosquito traps were conducted in southern Fulton County (Atlanta), GA, from June 9 to June 18 and June 24 to July 4, 2002, respectively. Total number of mosquitoes and number of each species captured during the tests were compared using a Latin square design. For the gravid trap infusion media, significant differences were found for total number of mosquitoes collected where sod > or = hay > or = hay side-by-side diluted hay > dilute hay side-by-side hay > or = oak > diluted hay. Only Aedes albopictus (oak), Culex quinquefasciatus (sod and both concentrated hay infusions), and Culex restuans (sod) were captured in significantly greater numbers using a particular infusion. Significant differences for the total number of mosquitoes collected were also observed in the commercial mosquito traps such that the gravid trap > ultra violet up-draft > or = Mosquito Magnet Pro > or = omnidirectional Fay-Prince trap with CO2 > up-draft CDC-style with CO2 > or = CDC-style with CO2. Significant differences in numbers collected among traps were noted for several species, including Aedes vexans, Aedes albopictus, Cx. quinquefasciatus, Cx. restuans, and Culex salinarius. Results from these field trap and infusion evaluations can enhance current surveillance efforts, especially for the primary vectors of West Nile virus and other arboviruses. PMID- 15532920 TI - The effectiveness of light, 1-octen-3-ol, and carbon dioxide as attractants for anopheline mosquitoes in Madang Province, Papua New Guinea. AB - The effectiveness of light, 1-octen-3-ol (octenol), carbon dioxide (CO2) and a combination of CO2 and octenol were compared as mosquito attractants using encephalitis vector surveillance traps in 2 villages in Madang Province, Papua New Guinea (PNG). Five species were collected, Anopheles koliensis, Anopheles farauti 2, Anopheles farauti 4, Anopheles longirostris, and Anopheles bancroftii. Light alone was not attractive to any of these species, and the attractiveness of octenol alone, though greater than light, was less than that of CO2 or the CO2 + octenol combination. With An. longirostris, the addition of octenol to CO2 resulted in a statistically significant increase in trap numbers; however, for the other species, any increase was not significant, and with An. koliensis and An. bancroftii, trap numbers were actually reduced when the CO2 + octenol bait was used. In PNG, the use of octenol alone would be effective in attracting more anophelines than if light alone was used; however, octenol by itself was not as effective as CO2. PMID- 15532921 TI - Development and survival of Anopheles gambiae eggs in drying soil: influence of the rate of drying, egg age, and soil type. AB - Little is known about the contribution made by the egg stage of African malaria vectors to the rapid rise in adult populations following the onset of seasonal rains. To examine this issue, we evaluated the viability of Anopheles gambiae eggs in drying soil in the laboratory. Survival data were collected from field caught mosquitoes kept in sandy loam soil and laboratory-reared colonies kept in sandy loam soil and black cotton soil. Under high, medium, and low soil-moisture regimes, egg viability declined sharply with increased duration of drying. Eggs remained viable in drying sandy loam soil for 1, 5, and 10 days, but not after 15 or 20 days. The most dramatic decline in hatching success occurred between drying days 1 (78-83% hatch) and 5 (20-23% hatch). In contrast, eggs reared in high moisture black cotton soil remained viable for up to 15 days. Furthermore, after 5 drying days, high-, medium-, and low-moisture soils averaged 59, 47, and 31% hatching success, respectively. We recovered unhatched eggs from sandy loam soils to examine the developmental status of the embryos. A majority of the unhatched eggs that were recovered from days 15 and 20 in sandy loam soils contained fully developed late-stage embryos. Thus, unhatched eggs completed embryonic development but probably died before receiving an appropriate hatching stimulus. Our results suggest that the absolute moisture content of the soil does not alone determine hatching success of anopheline eggs. Rather, soil moisture, together with the rate of drying, physiological factors associated with the age of the egg, and the type of soil in which the egg rests likely influence survival. PMID- 15532922 TI - Field evaluation of CDC gravid trap attractants to primary West Nile virus vectors, Culex mosquitoes in New York State. AB - A field study was conducted to evaluate two CDC gravid trap attractants available for the West Nile virus surveillance program in New York State (NYS). According to potential attractiveness, a common lawn sod in NYS, Kentucky bluegrass (Poa pratensis) infusion and a rabbit chow infusion were compared for attractiveness to primary West Nile virus vectors, Culex mosquitoes. Attractiveness of each infusion was measured by the number of adult mosquitoes caught in CDC gravid traps and the number of egg rafts laid in ovitraps. Both gravid trap and ovitrap studies demonstrated that lawn sod infusion with a 7-day incubation period had better attractiveness to Culex restuans/Culex pipiens than rabbit chow infusion with the same incubation period. Attractiveness of lawn sod infusions was increased as they became aged within a week's period. Lawn sod infusion also attracted more Ochlerotatus japonicus, a potentially important West Nile virus vector in New York. PMID- 15532923 TI - Evaluation of five trapping systems for the surveillance of gravid mosquitoes in Prince Georges County, Maryland. AB - Five mosquito trapping systems were evaluated in a large wildlife research center containing extensive and diverse mosquito-breeding habitat. The systems evaluated included Centers for Disease Control (CDC) New Standard Miniature Light traps with and without CO2 (dry ice), CDC gravid traps, partially open 1.8-m3 cages, and Fay-Prince traps baited with CO2. The first 4 trap systems were evaluated for 17 periods, while the Fay-Prince trap was evaluated on 5 trapping dates. Parameters recorded for each species were total catch, percent males, total females, and whether blood-fed/gravid. Fourteen of the 23 species caught yielded over 50 individuals in the 5 trapping systems (range of 71-2,524 specimens per species). Both light and CO2 were powerful attractants for 12 of the 14 commonly caught species. However, for most species, the majority of captured females were nongravid. Gravid traps caught fewer mosquitoes than did light- or CO2-baited traps, but the catch consisted of a higher percentage of gravid females. The open cages caught substantial numbers (>100 individuals) of 5 species, and for 2 species, this was the most productive trap. While light- and CO2-baited traps tended to catch few males or gravid females, the open cages caught an eclectic mixture of males, gravid females, and nongravid females, perhaps representative of the true percentages of each in nature. PMID- 15532924 TI - Yeast-generated CO2 as a convenient source of carbon dioxide for adult mosquito sampling. AB - A new, convenient method was developed to supply CO2 for mosquito sampling by using yeast, which converts sugar into CO2 and ethyl alcohol. The system could, at average, generate 32.4 ml/min of CO2 for at least 27 h. The total weight of the CO2 generated was estimated to be 94 g. The efficacy of yeast-generated CO2 as attractant for mosquitoes was significant, and the following 6 mosquito species were collected using yeast-generated CO2 traps from July to September 2003 in a residential area of southern and northern Yokohama City, Japan: Aedes albopictus (Skuse), Armigeres subalbatus (Coquillett), Culex halifaxii Theobald, Cx. pipiens pallens Coquillett, Ochlerotatus japonicus (Theobald), and Tripteroides bambusa (Yamada). Besides mosquitoes, various other insects were collected in the trap. Species compositions of insects collected in yeast generated CO2 traps and dry-ice-baited traps were compared. PMID- 15532925 TI - Use of polymerase chain reaction technique to confirm VecTest screening results in Plasmodium falciparum and Plasmodium vivax VK 210 laboratory-infected Anopheles stephensi mosquitoes. AB - We evaluated polymerase chain reaction (PCR) to confirm immunoassays for malaria parasites in mosquito pools after a failure to detect malaria with PCR during an outbreak in which pools tested positive using VecTest and enzyme-linked immunosorbent assay (ELISA). We combined VecTest, ELISA, and PCR to detect Plasmodium falciparum and Plasmodium vivax VK 210. Each mosquito pool, prepared in triplicate, consisted of 1 exposed Anopheles stephensi and up to 9 unfed mosquitoes. The results of VecTest and ELISA were concordant. DNA from a subset of the pools, 1 representative of each ratio of infected to uninfected mosquitoes, was extracted and used as template in PCR. All P. vivax pools were PCR positive but some needed additional processing for removal of apparent inhibitors before positive results were obtained. One of the pools selected for P. falciparum was negative by PCR, probably because of losses or contamination during DNA extraction; 2 remaining pools at this ratio were PCR positive. Testing pools by VecTest, ELISA, and PCR is feasible, and PCR is useful for confirmation of immunoassays. An additional step might be needed to remove potential inhibitors from pools prior to PCR. PMID- 15532926 TI - Susceptibility of Anopheles farauti to infection with different species of Plasmodium. AB - A colony of Anopheles farauti, originally from the island of New Britain in Papua New Guinea, was tested for its receptivity to infection with different species of Plasmodium in comparison with An. freeborni and An. stephensi. This colony adapted well to feeding on monkeys and was infected with New World and Old World strains of P. vivax and P. falciparum, P. ovale, P. cynomolgi, and P. brasilianum. PMID- 15532927 TI - Methods for studying the vector competence of Culex tarsalis for western equine encephalomyelitis virus. AB - Female Culex tarsalis fed heparinized chicken blood-western equine encephalomyelitis virus (WEEV) mixtures through a biomembrane feeder were compared with females fed sweetened blood-virus mixtures presented in pledgets or as hanging drops or to restrained chickens with natural or artificial viremias. Results indicated that sodium heparin did not adversely affect the infection of Culex tarsalis with WEEV. Overall advantages of the biomembrane system included 1) increased blood feeding frequency, 2) control of the infectious virus dose, and 3) greater or similar infection rates and body titers to females taking blood meals from viremic chickens. Anesthetizing females with triethylamine for in vitro transmission assessment using the capillary tube method produced results similar to immobilization using cold or CO2 + cold. Our research provided insight into tools useful to investigate the infection and transmission of WEEV by Cx. tarsalis. PMID- 15532928 TI - Efficacy of methyl bromide fumigation and insecticidal dips against Aedes eggs on lucky bamboo. AB - Importation of lucky bamboo (Dracaena sp.) stalk clusters has led to the accidental importation of Aedes albopictus in some countries. Current methods of fumigation using methyl bromide are harmful to live plants and thus are not acceptable. We exposed Aedes aegypti eggs to methyl bromide at rates of 32 g/m3 for 2 and 3 h and 48 g/m3 for 3 h, achieving complete control in 0/5, 1/5, and 5/5 replicates, respectively. Submersion for 1 min in aqueous solutions of 0.04% active ingredient (AI) lambda-cyhalothrin, 0.04% AI bifenthrin and carbaryl + malathion (both 0.1% AI) + petroleum oil (1% AI) provided 100% control of Ae. aegypti eggs laid on Dracaena stalk clusters. These methods, apparently harmless to plants, would have application in preventing the importation of exotic Aedes eggs on lucky bamboo shipments. PMID- 15532929 TI - Laboratory evaluation of the predation efficacy of native Australian fish on Culex annulirostris (Diptera: Culicidae). AB - The introduction and establishment of fish populations can provide long-term, cost-effective mosquito control in habitats such as constructed wetlands and ornamental lakes. The predation efficacy of 7 native Brisbane freshwater fish on 1st and 4th instars of the freshwater arbovirus vector Culex annulirostris was evaluated in a series of 24-h laboratory trials. The trials were conducted in 30 liter plastic carboys at 25+/-1 degrees C under a light:dark cycle of 14:10 h. The predation efficacy of native crimson-spotted rainbowfish Melanotaenia duboulayi (Melanotaeniidae), Australian smelt Retropinna semoni (Retropinnadae), Pacific blue-eye Pseudomugil signifer (Atherinidae), fly-specked hardyhead Craterocephalus stercusmuscarum (Atherinidae), firetail gudgeon Hypseleotris galii (Eleotridae), empire gudgeon Hypseleotris compressa (Eleotridae), and estuary perchlet Ambassis marianus (Ambassidae) was compared with the exotic eastern mosquitofish Gambusia holbrooki (Poeciliidae). This environmentally damaging exotic has been disseminated worldwide and has been declared noxious in Queensland. Melanotaenia duboulayi was found to consume the greatest numbers of both 1st and 4th instars of Cx. annulirostris. The predation efficacy of the remaining Australian native species was comparable with that of the exotic G. holbrooki. With the exception of A. marianus, the maximum predation rates of these native species were not statistically different whether tested individually or in a school of 6. Based on these data, M. duboulayi, H. compressa, and A. marianus warrant further investigation as biological control agents in pilot field trials. PMID- 15532930 TI - Laboratory and field evaluation of spatial repellency with metofluthrin impregnated paper strip against mosquitoes in Lombok Island, Indonesia. AB - Spatial repellency of a new multilayer paper strip impregnated with metofluthrin, a newly synthesized pyrethroid, was evaluated in the laboratory and in the field at Kerandangan, Lombok Island, Indonesia, with the use of cow- and human-baited double nets. Spatial repellency was observed in both cow- and human-baited collections. Metofluthrin treatment reduced mosquito collection by >80% during the 1st 4 weeks. However, repellency seemed to reduce with the loss of metofluthrin by evaporation within 6 wk after treatment. PMID- 15532931 TI - Comparative efficacy of IR3535 and deet as repellents against adult Aedes aegypti and Culex quinquefasciatus. AB - Arm-in-cage laboratory evaluations of 2 proprietary formulations of the mosquito repellents IR3535 and N,N-diethyl-3-methylbenzamide (deet; aqueous cream, hydroalcoholic spray) were made with 10 and 20% concentrations of each repellent. Also, 4 commercially available products containing IR3535 (Expedition insect repellent 20.07% active ingredient [AI], Bug Guard Plus with SPF30 sunscreen 7.5% AI, Bug Guard Plus with SPF15 sunscreen 7.5% AI, and Bug Guard Plus 7.5% AI) were tested. All comparisons were made on an equal formulation or concentration basis. Eight volunteers tested all formulations or products 3 times against laboratory reared, Aedes aegypti and Culex quinquefasciatus mosquitoes (6-10 days old). Products were applied to a forearm at the rate of 0.002 g/cm2. The other forearm was not treated and served as a control. Elapsed time to 1st and 2nd consecutive bite was recorded. Mean protection time (i.e., time to 1st bite) with proprietary formulations of IR3535 were comparable to those of deet, with 20% concentrations providing greater protection against Ae. aegypti (3 h) and Cx. quinquefasciatus (6 h). Mean protection time for commercial products containing IR3535 ranged from nearly 90 to 170 min for Ae. aegypti and 3.5 to 6.5 h for Cx. quinquefasciatus. Mean time to the 2nd bite was similar to time to 1st bite for each mosquito species, product, and formulation. PMID- 15532932 TI - Prey and size preference of Mesocyclops longisetus (Copepoda) for Aedes albopictus and Culex quinquefasciatus larvae. AB - Laboratory studies investigated prey choice of the adult copepod Mesocyclops longisetus for Aedes albopictus and Culex quinquefasciatus larvae. Prey size preference by this predator was tested within and between instar classes at 10 and 30 degrees C. Single copepod adults preferred to prey on 1st and 2nd instars regardless of whether either species was alone or combined. Generally, M. longisetus preyed more on Ae. albopictus than on Cx. quinquefasciatus when similar larval stages were present. Also more prey of both species were consumed at 30 degrees C compared with 10 degrees C. PMID- 15532933 TI - Larvicidal persistence of formulations of Bacillus thuringiensis var. israelensis to control larval Aedes aegypti. AB - After detection of resistance to the organophosphate temephos in populations of Aedes aegypti in Brazil, corncob granule (CG) and water-dispersible granule (WDG) formulations of Bacillus thuringiensis var. israelensis (Bti) were introduced in routine focal treatments. Larvicidal persistence and the influence of exposure to sunlight on VectoBac formulations of Bti were compared in 250-liter fiberglass water containers. Production of pupal Ae. aegypti in containers was used to indicate control. In untreated containers, survival of larvae was always above 95%. A temephos sand granule formulation used as reference treatment maintained 100% control throughout the 12-wk period in all situations. Under sunlight exposure, control dropped below the 90% level in the 2nd week after treatment at both dosages of VectoBac CG (1 and 2 g/50 liters) and VectoBac tablet (T) formulation at 1 tablet/100 liters. VectoBac T at 1 tablet/50 liters provided 2 wk of 100% control. VectoBac WDG at dosages of 1 and 2 g/500 liters provided 100% control for 3 wk. Without sunlight exposure (covered containers), VectoBac CG provided 9 wk of continuous 100% control and 5 wk of continuous 100% control, respectively, at 1 and 2 g/50 liters. The VectoBac T formulation at both dosages initially provided 2 wk of 100% control. After this period, the control level fluctuated between 96 and 100%. VectoBac WDG provided continuous 100% control for 7 wk for the lower dosage and for 6 wk for the higher dosage. At both dosages of WDG, 100% control was achieved in 11 wk out of the 12-wk period. PMID- 15532934 TI - Aedes fumidus and Aedes amesii in mangrove forests of India--two new country records. AB - We report the first records of Aedes fumidus and Ae. amesii from India based on collections made in mangrove forests of Bhitarkanika in Orissa, Sunderbans, in West Bengal, and Andaman and Nicobar Islands. Collection details include date, location, material examined, larval habitat, and associated species. PMID- 15532935 TI - Resting behavior and malaria vector incrimination of Anopheles stephensi in Goa, India. AB - Malaria in Goa, India, has been endemic ever since an outbreak occurred in 1986. Anopheles stephensi Liston has always been suspected as a malaria vector in this area. Due to lack of knowledge on its resting behavior, sufficient adult females could not be collected and incriminated as vectors in the past. In this study mosquito collections were conducted in three endemic urban and suburban areas of Goa. In well-built houses, 67 h of collections did not yield a single An. stephensi mosquito, although other species were encountered. However, collections in construction sites and workers' huts for 151 h yielded, besides other mosquito species, 38 An. stephensi females resting in 15 types of sites at a height varying from 30 cm to 2.4 m. Of the 37 of these mosquitoes tested for the presence of circumsporozoite protein (CSP) by an ELISA technique, 1 was found to be Plasmodium falciparum CSP positive. PMID- 15532936 TI - Changes in distribution and abundance of mosquito populations in an ecological research tract over a 35-year period. AB - Marked changes in species and numbers of mosquitoes have occurred over a 35-year period. Changes relate to alteration of available larval development sites by beaver (Castor canadensis), variation in winter and spring precipitation, and differences in feeding behaviors. PMID- 15532937 TI - A readily constructed lard-can trap for sampling host-seeking mosquitoes. AB - Although lard-can traps have been used for sampling host-seeking mosquitoes for at least a half-century, the materials from which they originally were constructed no longer are available. We therefore devised a method for constructing such devices from parts available in the ventilation industry. These traps, baited with birds and mounted near the tops of trees, were employed to monitor the host-seeking activity of Culex spp. mosquitoes. Lard-can traps, constructed in this manner, are economical and sturdy and effectively sample the Culex mosquitoes that appear to perpetuate West Nile virus in North America. PMID- 15532938 TI - Cocoa pod (Theobroma caco)--a potential breeding habit of Aedes albopictus in dengue-sensitive Kerala State, India. AB - Aedes (Stegomyia) albopictus (Skuse), a potential dengue vector, particularly in the foot-hill areas of the Western Ghat region of Kerala State has been found breeding for the first time in cocoa pods (Theobroma cacao) in India. Breeding in cocoa pods is considered an opportunistic adaptive behavior of the mosquito to maintain density in the absence of common breeding sites, especially latex collecting cups in rubber plantations, during the monsoon periods. Vector breeding in cocoa pods was observed up to 8 m from the ground, and in both hanging and grounded pods, whose endocarp had been devoured by rodents. Ae. albopictus breeding in cocoa plant is discussed in relation to accentuating disease transmission risk. PMID- 15532939 TI - The discovery of West Nile virus in overwintering Culex pipiens (Diptera: Culicidae) mosquitoes in Lehigh County, Pennsylvania. AB - In February 2003, a pool of 6 Culex pipiens mosquitoes collected in Lehigh County, PA, tested positive for West Nile Virus (WNV). West Nile viral RNA was detected by reverse transcriptase-polymerase chain reaction. This is the first time in Pennsylvania and the third time in North America that WNV has been found in overwintering adult mosquitoes. PMID- 15532940 TI - Influence of the blood meal source on the biology of Meccus longipennis (Hemiptera: Reduviidae) under laboratory conditions. AB - Influence of the blood meal source on life cycle, mortality, and fecundity of 2 cohorts of recently colonized Mexican Meccus longipennis, fed on hens (H-cohort) or rats (R-cohort) were evaluated in laboratory conditions. One hundred twelve nymphs (56%) (H-cohort) and 102 nymphs (51%) (R-cohort), completed the cycle. The average time from Nymph I to adult was 209+/-41 days (H-cohort) and 239+/-28 days (R-cohort), taking an average of 1.8 (H-cohort) and 1.9 (R-cohort) blood meals per nymphal stage (range 1-6). The average span in days for each stage from the H cohort was 20.8 for Nymph I, 24.5 for Nymph II, 38.8 for Nymph III, 56.1 for Nymph IV, and 72.5 for Nymph V, and it was 20.2 for Nymph I, 23.1 for Nymph II, 43.2 for Nymph III, 68.8 for Nymph IV, and 75.4 for Nymph V from the R-cohort. The mortality percentage was 44% (H-cohort) and 49% (R-cohort). The average number of eggs laid per female in a 9-month period was 484.1 (range 351.1-847.8) in the H-cohort, whereas the average number of eggs was 442.3 (range 288.5-720.5) in the R-cohort. No significant differences (P > 0.05) were recorded among cohorts fed on the studied blood meal sources, different from most previously studied Triatominae species, perhaps due to a high degree of association of M. longipennis with chickens and hens as much as with mammals under natural conditions on human dwellings. PMID- 15532941 TI - The control of medical doctors in Belgium. AB - First the structure and purpose of three institutions of medical control in Belgium are presented (the Order of Physicians, the provincial medical committees, the medical control service). Secondly, data with respect to their actual functioning are discussed. Finally the present obstacles to effective and efficient self-regulation and control of medical doctors are treated on the basis of our research findings. PMID- 15532942 TI - Medical litigation in cosmetic plastic surgery. AB - AIM: This article aims to highlight issues related to malpractice in plastic surgery and to point out the importance of good understanding of the law and the value of a patient's written informed consent as measures of professional protection. METHODS AND MATERIAL: Search of relevant literature from PubMed. RESULTS: The demand for cosmetic plastic surgery increases despite the increasing cost, in contrast to other traditional goods for which demand typically declines as price increases. Cosmetic plastic surgery has moved beyond the stage of being an exclusive privilege of the rich and famous. Nevertheless, cosmetic plastic surgery is one of the medical specialties exposed to a substantially high risk of malpractice claims. Most malpractice claims in cosmetic plastic surgery are not consequences of technical faults but because of inadequate patient selection criteria and lack of adequate communication between patient and surgeon. Proven efficient training, careful utilization of computer imaging techniques in association with the adoption of simple precautions and guidelines and adequate communication along with a completed patient's consent form are important essentials in case of medical litigation. CONCLUSIONS: In today's litigious society, maintenance of high standards in daily practice with continuous training and appropriate documentation of every procedure are all a sufficient defense of the plastic surgeon in case of medical litigation. Written patient's informed consent remains an integral part of the communication between physicians and patients, and importantly is facilitating professional protection. PMID- 15532943 TI - Late complications after medical treatment--malpractice or fate? AB - During the past decade, interest in medical malpractice has increased both by the public and the authorities. Malpractice is likely to be considered if an adverse effect occurs shortly after medical intervention, but complications might lead to impairment or death of the patient even years after surgical treatment. Two unusual cases of such late complications are described. In the first case, the patient died from a complication, which was due to an unnoticeable malfunction of a surgical device. The second case concerns a retained surgical towel, which had remained within the abdominal cavity for 25 years without causing any complaints and had no influence on the cause of death. Medico-legal autopsy is an indispensable basic tool in the assessment of medical malpractice. PMID- 15532944 TI - Medical malpractice: managing the risk. AB - STUDY OBJECTIVE: This is an attempt to present an analysis of the literature examining objective information concerning the likelihood of medicolegal errors as it applies to current medical practice. Hopefully this information will be synthesized to generate a cogent approach to manage risk in emergency medicine. METHODS: Articles were obtained by an English language search of MEDLINE from January 1976 to July 2003. This computerized search was supplemented with literature from the author's personal medicolegal collection of peer review articles. This information was presented in a qualitative fashion. RESULTS: There was a steady increase in both the incidence and the recovery amount of verdicts involving general malpractice litigation. There are clearly high-risk emergency medicine categories responsible for most malpractice events, involving such commonly encountered conditions such as chest pain, abdominal pain, pediatric fever, central nervous system (CNS) bleeding, and abdominal aortic aneurysm (AAA). Interestingly, there is a second peak of more minor emergencies, specifically wounds with neglected foreign bodies and missed fractures. Clearly, the largest dollar amount recovery still involves chest pain with subsequent missed transmural myocardial infarction (MI). Interestingly, there does not appear to be a strong correlation between adverse events, outcome and medicolegal risk. Likewise, there does not appear to be a strong correlation between socioeconomic status and a propensity to sue, but there were some defined links with physician profiles involving past malpractice history, as well as prior adverse relationships or communication skills to subsequent claims. Interestingly, a significant association appears to be advertising placed by local law offices seeking to provide services. Lastly in the emergency medical services (EMS) realm, the single strongest correlate to malpractice was the likelihood of an ambulance accident and not related to care delivered itself. CONCLUSION: The current emergency medicine medicolegal dilemmas are a complex interaction of both patient and physician factors specifically targeting several disease categories and damage claims. Awareness of these issues can help to minimize subsequent medicolegal risk and improve patient care. PMID- 15532945 TI - The doctor's dilemma: caregiving and medicolegal evidence collection. AB - This paper reports on findings from a population-based study of sexual assault physicians who are responsible for both the health care of sexually assaulted persons and for conducting forensic medical examinations on them for the purpose of collecting corroborative evidence to aid in legal proceedings. Self administered questionnaires were distributed to the 100 physicians attached to sexual assault care and treatment centres across the province of Ontario, Canada. The questionnaires were focussed on exploring their relative commitment to two assessment priorities: the collection of forensic evidence for the courts and the provision of medical care. While most of the 31 physicians replying expressed satisfaction with their participation in the legal arena, a substantial proportion indicated that although they believed medical care to be the most important part of their work, evidence collection and court appearances were the most time-consuming. In addition, over four-fifths admitted to having deviated from the standard protocol used for evidence collection, based on the belief that a particular procedure or information item might either emotionally or legally harm the victim or her/his case. We suggest that the implications of these findings point to a possible ambivalence towards the evidentiary dimension of the dual role of the sexual assault physician. PMID- 15532946 TI - Expanding the physician's duty of care: a duty to recontact? AB - This article addresses the ethical and legal issues surrounding the possibility of a "duty to recontact" patients concerning new genetic developments. The text examines the rationale of this concept, the conditions required for the creation of such a duty and who should be bound by it. While this "duty to recontact" has yet to be developed, the predictive familial dimensions of genetic information, however, may lead policy makers to favour such a duty. Professional organizations should begin to assess the implications of the genetic revolution on patient management and physicians' continuing education. PMID- 15532947 TI - Telemedicine and informed consent. AB - Though pursuing robotic technology offers a solution to geographically or economically isolated patients, it also causes new problems with regard to the safeguarding of patients' rights, including those relating to informed consent. Questions that need to be answered concern the content of the informed consent requirement; the form of the consent; whose responsibility is it to gain the consent, etc. In this paper we will discuss all these topics and take a closer look at international aspects, since telemedicine can also be performed across state boundaries. PMID- 15532948 TI - Is bioethical training useful in preparing doctors to take decisions in the emergency room? AB - We study the decisions taken in five real cases by 178 doctors working in hospital emergency services and compare their decisions with those proposed a reference group composed of professionals with a master's degree in bioethics. The findings of our study point to an increased ability to take difficult decisions in critical situations involving the use of life-support measures in the emergency room. The group of professionals chosen as "gold standard", despite lacking the training and clinical preparation of emergency doctors, made decisions that were technically very close to the most suitable. In this respect, an adequate ethical training facilitated the taking of decisions that required the involvement of personally held values, underlining the need for such training in the case of professionals who will work in hospital emergency services. PMID- 15532949 TI - From the dead to the unborn: is there an ethical duty to save life? AB - In this paper I examine the question whether physicians have a legal and ethical duty to sustain pregnancies of women who die during the first or second trimester by the delivery of their fetuses. One ground for such a duty, on which I am focusing, is the duty of "special relationship" between the mother and the fetus. In my paper, I claim that the special relations the pregnant woman and the fetus have do provide such a moral duty. This moral duty derives from the special and intimate relationship between the mother and the fetus, which has two considerations that support it: the uniqueness of the fetus, and the mother's understanding and acknowledgement of such uniqueness. However, I argue that when the mother is dead, the nature of the relationship changes, as she is no more aware of her fetus and her relationship with it. Thus, the ethical duty of the mother to save her fetus' life should be declined upon the end of the relationship between the mother and her fetus with the mother's death. I support my argument by analyzing the special relationship between the mother and the fetus from four related ethical theories: ethics of relationships, responsibilities to society, ethics of families, and the ethics of care. By discussing these ethical theories, I show how responsibility to society in general, and to social entities, like families, in particular, constitute a moral duty towards the fetus, which, as aforesaid, no longer exists upon the pregnant woman's death. In addition to being social entities I further show how the intrinsic values of families play an important role in forming such a moral duty. Nevertheless, I argue that such an instrumental duty that enables the establishment of families no more exists as the pregnant woman is no more socially and morally part of the family she belonged to while alive. I strengthen my argument by applying ethics of care, and by analyzing the practical conclusion I arrived at from a religious perspective. PMID- 15532950 TI - Lay perspectives on ethics in epilepsy. PMID- 15532951 TI - Epilepsy, ethics, consent and emergency care. PMID- 15532952 TI - Comparison between two legal indications for compulsory outpatient treatment in Israel. AB - The aim of the present study was to compare the outcome of two legal indications for Compulsory Outpatient Treatment (COT) in Israel (a) as an alternative to compulsory hospitalization (CH) and (b) as a follow-up to CH. Demographic, clinical and legal measurements based on 326 COT orders issued in the Jerusalem and Southern district of Israel were compared according to the two legal provisions for COT contained in the Treatment of Mental Patients Law (1991). The COT order was found to be more effective when used as an alternative to CH (53.1%) than when it was used following CH (38.8%), p = 0.002. In the former case, there were fewer schizophrenic patients, fewer hospitalizations, fewer visits to psychiatric emergency services and longer remissions. Our results point to the influence of the legal indications on the efficacy of the COT order and help define the target population. PMID- 15532953 TI - Characteristics of district psychiatric committee decisions regarding discharge from compulsory hospitalization. AB - In 1991 the authority of psychiatric committees responsible for judicial decisions concerning mentally ill individuals hospitalized compulsorily was greatly extended. The committees' work procedures have gradually been established despite lack of binding rules and regulations, often evolving from the dynamics of the field. METHODS: The study population included 92 hospitalized patients. The examination was performed by administering a questionnaire, reading a protocol of the latest committee hearing and perusing the patient record. RESULTS: The committee tends to decide gradually about discharge from hospitalization and sometimes to include it as part of a rehabilitation process. The reasoning leading to discharge is generally from the field of law. CONCLUSIONS: Tension between medical and legal approaches often underlies psychiatric committee work. Generally, the committee tends to accept the position of the treating physicians. This may indicate that the two approaches are drawing closer to one another and that they have learned to work together cooperatively and fruitfully. PMID- 15532954 TI - The relationship between memory and the recall of specific details. AB - When eyewitnesses are exposed to an event, certain details tend to capture their attention more than other details, which ensures the better recall of the details that were observed. As little research has focussed on the relationship between certain biographical variables (occupation, age, gender, race and retention period) and specific event details, the present study attempts to shed some light on this area. A sample of 295 participants was obtained from 12- to 14-year-olds, university students, the public and Police College students that were divided into a short-term and long-term memory group. The details that were focussed on in the questionnaire were divided into obvious physiognomic, less pronounced physiognomic, clothing, circumstantial, and action-related details. Between the short-term and long-term memory groups it was found that significant differences were indicated for only certain items related to obvious physiognomic details, clothing details, circumstantial evidence and action-related details. For all the different biographical groups significant differences were found for certain items belonging to the different groups of details. PMID- 15532955 TI - Law, ethics and professional guidelines in medical research. AB - The volume of law and other regulation in medical research has expanded enormously within the last five years. Researchers need an awareness of how these legal instruments are hierarchically related, so they know which regulation to apply where there is a conflict of norms. They also express concern on whether there is any space left for ethical and professional guidelines in the light of all this formal regulation. This paper addresses these two issues. It is argued that when drafting new legislation or guidelines, room must be left for responsible individual decision-making. PMID- 15532956 TI - Abortion policy in Turkey: current ethical attitudes. AB - The authors outline the development of abortion law in Turkey, and relate ethical attitudes to current practice, especially in connection with policies of population control, and the institution of secular law in a previously Islamic legal framework. PMID- 15532957 TI - African ethics and voluntary euthanasia. AB - This paper outlines the relationship between euthanasia and its ethical norms and practices in a part of West Africa. The various sub-types of euthanasia are described in detail, parallel with the role of African ethical theories in determining their relevance. The author discusses the implications of this approach relative to the social and economic state of African communities. PMID- 15532958 TI - Euthanasia in the Netherlands: experiences in a review committee. AB - Since 1999 reported cases of euthanasia in The Netherlands are examined in five regional review committees. In these committees a lawyer, an ethicist and a medical doctor investigate whether the cases reported are in correspondence with the so-called criteria for careful practice. Issues to be investigated include the patient's suffering, the nature of the patient's request, the consultation by another doctor and the way in which the termination of life has been performed. Since April 2002 the judgment of the committees is final. This paper describes the working of the review committees. It also discusses the relevance of experiences in the committee for issues of social and public debate, such as the role of the consulting doctor, the relation between euthanasia and palliative care, and the place of patient autonomy. PMID- 15532959 TI - [Criteria for assessing severely hot environments: from the WBGT index to the PHS (predicted heat strain) model]. AB - BACKGROUND: The present study deals with the main methods for assessment of hot environments: i.e., WBGT, SWreq and PHS. It is stressed how the WBGT index, which is strictly empirical, although a very practical tool for the assessment of the hot environments, can only be used for a rough evaluation of heat stress, and especially for a not very high metabolic rate (M<175 W/m2). On the contrary, the SWreq method, which is based on both subject-environment heat exchange and the effect of clothing, allows a better assessment of the work situation with a general reduction of the exposure limits with respect to WBGT, especially in non uniform environments (ta not equal to tr). However, it should be noted that application of SWreq is required by the ISO standard 7243 when the WBGT limit values are exceeded. METHODS: In this study interest was extensively focused on the "Predicted Heat Strain" method, highlighting via a special software the differences in heat stress assessment related to this new approach, which will be adopted by the ISO in the next revision of standard 7933. RESULTS: The PHS method, unlike SWreq, allows the prediction of the time-response of the main physiological variables of interest (i.e., skin temperature, core temperature and sweat rate). Moreover thanks to better modelling of heat exchanges, the PHS method allows account to be taken of both movement and clothing effects, resulting in even more reduced exposure. PMID- 15532960 TI - [Occupational asthma: role of airway inflammation and remodelling in persistent respiratory symptoms and bronchial hyper-responsiveness]. AB - BACKGROUND: Until the late 1970's occupational asthma (OA) was considered reversible once patients were removed from exposure. Unfortunately, respiratory symptoms and non-specific bronchial hyper-responsiveness (NSBH) persist in about two-thirds of patients for years after removal from the offending agent. OBJECTIVES AND METHODS: This review focuses on the role of airways inflammation and remodelling in persistent respiratory symptoms and NSBH after cessation of occupational exposure. RESULTS: Even though cessation of exposure does not always result in remission of OA, symptoms, airways calibre and NSBH do improve in many patients. Although improvements in FEV1 and NSBH tend to reach a plateau 1-2 years after workers leave exposure, reversing NSBH may take much longer and respiratory symptoms and NSBH can persist in subjects removed from exposurefor >10 yrs. Long-term treatment with inhaled corticosteroids (ICS) induces a small but significant improvement in respiratory symptoms and in quality of life and a decrease in NSBH. Prolonged exposure and respiratory symptoms, marked airway obstruction and NSBH, high total cell, eosinophil and neutrophil counts in bronchoalveolar lavage fluid, a strong reaction during specific inhalation challenge, and delayed treatment with ICS have been identified as prognostic factors of unfavourable outcome. If exposure persists, OA tends to deteriorate in many patients but regular long-term treatment with ICS and long-acting beta2 agonists seems to stabilize the outcome. Soon after the last exposure inflammatory cell infiltrates, including eosinophils, and increased thickness of sub-epithelial collagen have been observed. When time since removal from exposure was longer, persistence of respiratory symptoms and NSBH was associated with airway inflammation, remodelling and hypersensitivity to the offending agent. Thickness of sub-epithelial collagen and specific airway sensitivity were reduced after prolonged non-exposure to isocyanates, although NSBH and airway inflammation persisted. CONCLUSIONS: Pathologic features are similar in OA and non-occupational asthma. The main factors of favourable outcome are early removal from exposure and a mild airway obstruction and NSBH at diagnosis. Persistence of airway inflammation years after removal from exposure suggests this process may become independent of the offending agent. The role of remodelling on persistence of OA needs to be clarified further. PMID- 15532961 TI - [Hand-arm vibration syndrome and upper limb disorders associated with forestry work]. AB - BACKGROUND: Occupational exposure to hand-transmitted vibration in forestry workers is associated with an increased risk for vascular, neurological and musculo-skeletal disorders of the upper limbs. OBJECTIVES: To carry out a cross sectional study of the hand-arm vibration syndrome and soft-tissue disorders of the upper limb in a group of forestry workers employed in the Forestry Service of the Province of Trento (Italy). In the forestry worker group, usage of anti vibration chain-saws was intermittent over a typical work year (16 weeks/yr, on average). METHODS: To investigate vascular, neurological and musculo-skeletal disorders of the upper limbs, the forestry workers (n=159) and a control group of manual workers, unexposed to hand-transmitted vibration, employed in the same Forestry Service (n=146) underwent a structured medical interview and a complete physical examination. The clinical diagnoses of vibration-induced white finger (VWF) and carpal tunnel syndrome were made according to internationally recognised consensus criteria. Occupational exposure to hand-transmitted vibration was assessed according to the recommendations of the International Standard ISO 5349-1 (2001). RESULTS: The forestry workers showed an increased prevalence of peripheral sensory-neural disturbances (33.3%), musculo-skeletal disorders of the upper limbs (37.7%), and carpal tunnel syndrome (21.4%) compared to those observed in the control group. There was no significant difference in the prevalence ofRaynaud' sphenomenon between the forestry workers (6.3%) and the controls (4.1%). After adjustment for confounding factors (age, body mass index, tobacco and alcohol consumption), a significant association was observed between peripheral neuropathies (peripheral sensory-neural disorders, carpal tunnel syndrome) and several indices of vibration exposure such as 8-hr energy equivalent frequency-weighted acceleration [A(8) in m/s2 r.m.s.], duration of exposure (years), and lifetime vibration dose (m2/s4 hr). An excess, although not significant, risk for VWF was observed only in the forestry workers with A(8) > or = 4 m/s2 r.m.s.. In the forestry workers, there was no significant association between VWF and sensory-neural disorders, and between VWF and carpal tunnel syndrome. This finding seems to support the hypothesis that the vascular and neurological components of the hand-arm vibration syndrome develop independently of each other. Within the forestry worker group, the variable "years of tool usage" showed the strongest association with peripheral sensory-neural symptoms, carpal tunnel syndrome, and soft-tissue disorders of the upper limbs. It is likely that in the forestry workers the variable "years of tool usage" represents an overall index of duration of exposure to several adverse ergonomic and mechanical risk factors. CONCLUSIONS: The results of this study suggest a tendency for a decrease in the occurrence of VWF among forestry workers, and this finding seems to be associated with the use of anti-vibration chain saws, the reduction of exposure duration, and the improvement of work organization. On the other hand, the forestry workers showed an increase in the occurrence of peripheral sensory neuropathies, carpal tunnel syndrome, and upper-extremity musculo-skeletal disorders. This finding seems to support the view that there is epidemiological evidence for a positive association between exposure to a combination of risk factors (segmental vibration, forcefulness, awkward posture) and the occurrence of soft-tissue disorders of the upper limb in working populations. PMID- 15532962 TI - [Methodology for assessing occupational dermal exposure]. AB - BACKGROUND: Dermal risk assessment of exposure to chemicals is difficult due to the lack of standardisation of the methods used in exposure quantification; further limits concern the extent of skin contamination and especially dermal absorption of different substances. To solve these problems many authors proposed conceptual models founded on theoretical assumptions and experimental data, but their use in occupational settings revealed many difficulties. A possible alternative is the use of a check-list that permits evaluation of occupational dermal contamination hazards. AIMS: To propose a simplified method for dermal risk assessment. METHODS: Dermal contamination risk evaluation, chemical hazard quantification and duration of exposure are the basic features of the method. The first step assesses working conditions, use of protective devices, skin contamination directly observed, personal hygiene of workers: all information, collected by a trained observer, is scored according to literature data and/or to authors'personal experience. Chemical hazard quantfication is obtained from safety data sheet evaluation (R phrases classification and chemical-physical characteristics of the chemical in use). The method allows a preliminary dermal exposure assessment to be made, subdivided on hazard bands. RESULTS: The method permits a rapid dermal exposure assessment to be made and suggests intervention measures to decrease dermal risk. CONCLUSIONS: A check-list is proposed that can be a useful and easy method to assess dermal exposure to chemicals, particularly in small- and medium-sized enterprises. PMID- 15532963 TI - [The OCRA method: updating of reference values and prediction models of occurrence of work-related musculo-skeletal diseases of the upper limbs (UL WMSDs) in working populations exposed to repetitive movements and exertions of the upper limbs]. AB - BACKGROUND: The paper considers a database of old (already published) and new data concerning 23 groups of workers (Total number of subjects examined=5373) with different levels of exposure to repetitive movements of the upper limbs: for all these groups data were available regarding exposure indexes (OCRA index and Checklist "OCRA" score) and clinically determined UL-WMSD outcomes (PA=Prevalence of workers Affected by one or more UL- WMSDs; PC=Prevalence of single diagnosed Cases of an UL- WMSDs). OBJECTIVES: Using these data, the paper aimed at presenting and discussing the results obtained in order to estimate: new critical values of OCRA index for discriminating different exposure levels (green, yellow, red areas); new prediction models of expected PA and PC in exposed populations based on exposure indexes. METHODS: New critical values of the OCRA index (and, consequently, of the checklist score) were estimated by an original approach in which data of the effect variable PA in a reference population not exposed to the specific risks were combined with the regression function between OCRA and PA, as resulting from the 23 available groups. RESULTS: The resulting critical values and the consequent classification system of the OCRA index and of the checklist score are synthetically reported in the following table: [table: see text]. The best simple regression functions between exposure indexes (OCRA; checklist) and health outcome variables (PA; PC) were then sought, in order to obtain prediction models of effects starting from exposure. The following were the main prediction models derived from the available set of data (standard error of b in brackets): [formula: see text]. Finally, a multiple regression model was computed for estimating PA (Y) based on OCRA index and gender structure of the group (SEXRATIO=n. females x 100/n. total) with its 5 degrees and 95 degrees percentiles (in brackets); the resulting model was. Y = 2.02 (1.72-2.32) x OCRA + 0.075 (0.035-0.115) x SEXRATIO. This model showed a very high association between the two independent variables and the effect variable (PA) (R2=0.96). DISCUSSION: Discussion of the results obtained considers their intrinsic limits, as they are based on prevalence studies, and also suggests due recommendations and caution in the use of the proposed classification system and prediction models when the OCRA methods are applied for the evaluation of occupational risk associated with repetitive movements of the upper limbs. PMID- 15532965 TI - [Education and assistance: activity necessary for the training in the specialization of occupational medicine]. PMID- 15532964 TI - [Malignant mesothelioma of the pleura in a worker with brief atypical exposure to chrysotile asbestos]. AB - BACKGROUND: The appearance of malignant mesothelioma in workers exposed to asbestos dusts even for relatively short periods of time is amply demonstrated in the literature. The workers however were usually exposed to amphiboles in jobs well known as involving exposure, that are characterized by high levels of dusts. OBJECTIVES: To describe a clinical case of pleural mesothelioma that occurred in a patient exposed to chrysotile, in a job (driver) that cannot be directly linked to such high exposure, and which moreover he only did for a few months. METHODS: The clinical history was reconstructed by analyzing the clinical files of the hospital admittances from May 2002 to August 2003, during which the patient underwent radiological examinations (chest x-rays, chest and abdomen TC), cytological examination of the pleural fluid, videothoracoscopic surgery with histological examination (including immunohistochemical coloration) of the tissue taken in biopsy. The job history, as well as any possible non-occupational exposures to asbestos, was examined via a standardised questionnaire, which the patient himself answered, as used in the Lombardy Mesothelioma Register, in operation at the "Clinica del Lavoro" in Milan. RESULTS: Examination of all clinical files confirmed the diagnosis of malignant pleural mesothelioma. Analysis of job history was found appropriate for defining as certain an occupational aetiology due to inhalation of asbestos fibres which occurred for few months as a truck driver in a chrysotile mine. PMID- 15532966 TI - Organ retention: learn the lessons but please move on. PMID- 15532967 TI - The Mirena Intrauterine System--reliable contraception for the 21st century. PMID- 15532969 TI - Prescription patterns in the elderly population--"new" versus "old" medical card holders. AB - INTRODUCTION: In July 2001, the GMS Scheme was extended to the whole elderly population aged 70 and over, permitting access to medical care services free of charge. We undertook a study to compare prescribing patterns between those relatively affluent people referred to as "new" over 70s with those relatively deprived people referred to as "old" over 70's, over the 18 months since the introduction of the scheme. Patients who had received a prescription for antibacterials, diuretics, psycholeptics, psychoanaleptics, statins, b-blockers, antithrombolytics, antianaemic drugs and drugs for obstructive airway diseases, were identified over the 18 month period using the GMS database. We also compared the average defined daily dose (DDD) received per month for each of the therapies above. RESULTS: All therapies directed at treatment were significantly more likely to be prescribed to the old over 70's, such as the vasodilators (OR=1.59, 1.51-1.67), peptic ulcer drugs (OR=1.37, 1.33-1.43) and the antibacterials (OR=1.37, 1.33-1.41) with the exception of those associated with cardiovascular prevention; statins (OR=0.88, 0.85-0.92), beta-blockers (OR=0.95, 0.92-0.98) and antithrombolytics (OR=0.96, 0.93-0.99). The old over 70's received more prescriptions with a higher than the average DDD per month for vasodilators, diuretics, and drugs for obstructive airway disease. This study highlights a potential inequality in prescribing in primary care, evident following a change in the health policy in Ireland. These results suggest that differences in socio economic status (such as income) and morbidity may be associated with differences in prescribing by GPs. PMID- 15532968 TI - Smoking, alcohol & illicit drug use among young people in a health board region in 1997 and 2002: a comparative study. AB - The objectives of this study were to document and compare patterns of licit and illicit drug use among adolescents in a health board region in 1997 and 2002. 1516 students in 1997 and 1426 in 2002, randomly selected from post-primary schools in the region, completed a questionnaire, incorporating items related to smoking, alcohol and illicit drug use. Lifetime smoking prevalence in 2002 (50.8%) showed a statistically significant decrease from 1997 (57.1%). There was also a statistically significant decrease in regular smoking (1997 - 30.7%; 2002 18.2%). There was no significant change in regular alcohol consumption (57.3% - 1997; 53.7% - 2002). However, there was an increase in binge drinking at weekends and reports of feeling drunk more than ten times (24% - 1997; 27.2% - 2002). In 2002, 41.2% had ever taken an illicit drug, a statistically significant increase from 1997 (34.9%); 15.1% were regular users, also a statistically significant increase from 1997 (11.9%). These findings highlight that while the misuse of illicit drugs has increased, smoking has significantly declined since 1997. However, alcohol continues to be a major problem within this age-group. This study has implications for the implementation of services and strategies aimed at reducing smoking, alcohol and drug use within this population. PMID- 15532970 TI - The evaluation and treatment of female urinary incontinence--a comparison of clinical practice in the Republic of Ireland with the recommendations of the International Continence Society. AB - In 1998 the International Continence Society (ICS) developed a set of recommendations for the diagnostic evaluation and treatment of urinary incontinence. The aim of this study was to determine to what extent current clinical practice in the Republic of Ireland correlated with the ICS recommendations. We personally interviewed 100 gynaecologists at units around Ireland. Participants were presented with two clinical scenarios, one described a patient with predominantly genuine stress incontinence, the other described a patient with an overactive bladder. Ninety-five (95%) requested a midstream urine sample for culture and sensitivity, and 74 (74%) considered urodynamics an appropriate initial investigation for a woman with stress incontinence. Physiotherapy was recommended as a first line treatment for stress incontinence by 76 (76%). Burch colposuspension, chosen by 55 (55%), was the most common first line surgical procedure. Other first line surgical procedures were TVT [31(31%)] Marshall-Marchetti-Krants procedure [5(5%)], anterior colporrhaphy [4 (4%)] and a variety of other procedures [5 (5%)]. When considering the initial investigation for a woman with an overactive bladder, 95 (95%) asked for a midstream urine sample for culture and sensitivity and 85 (85%) requested urodynamic investigations. Initial management of this condition included anticholinergic therapy alone [57 (57%)], anticholinergic therapy combined with bladder retraining [36 (36%)], bladder retraining alone [5 (5%)], and cystoscopy and bladder distension 2 (2%). Seventy-six (76%) felt that cystoscopy was an appropriate investigation for a woman with symptoms of an overactive bladder who has failed to respond to initial therapy. The study reveals a significant degree of diversity in the evaluation and management of patients who present with symptoms of urinary incontinence. It also highlights a number of areas where current clinical practice deviates from the recommendations of the ICS. In particular, there is a high and inappropriate use of urodynamics in the initial management of these patients. PMID- 15532971 TI - The value of the perinatal and neonatal autopsy. AB - The postmortem historically has been considered a valuable diagnostic exercise which contributes to medical knowledge. Despite this, there has been a significant reduction in autopsy rates throughout the developed world. This audit was a retrospective study of autopsy reports of stillbirths and neonates [corrected] greater than 500 grams over a five year period from 1995 to 1999. The audit was performed to assess the impact of autopsy on the current practice of perinatal medicine. The audit compared the final pathological diagnosis to the clinical diagnosis. The pathological diagnosis was categorised as (I) diagnostic, (II) confirmative, (III) unexplained. The recurrence risk estimates as a result of necropsy were also identified, as were any additional findings that were felt to be relevant but did not belong to any of the above categories. A total of 262 perinatal deaths (including stillbirths) and neonatal deaths greater than 500 grams birth weight were recorded during this period. The autopsy rate was 81%. The 213 autopsies performed were assessed, of which 76 (36%) were found to be diagnostic, 108 (51%) confirmatory, and 29 (13%) were unexplained or revealed no new findings. Change in recurrence risk estimates was identified in 24 (11%) and additional relevant information was obtained in 38 (18%). There were a number of cases where an unexpected diagnosis was made as a result of autopsy; these diagnoses included a respiratory chain disorder in a twenty nine week infant, and an occult necrotising enterocolitis presenting with severe haemolysis post transfusion in a preterm infant. The perinatal post mortem examination remains an indispensable part of clinical management. It contributes to medical education and quality assurance. It can aid in the identification of inheritable diseases and provide information for accurate parental counseling. PMID- 15532972 TI - Firework injuries presenting to a national burn's unit. AB - The sale to the general public of fireworks is illegal in Ireland. However, many fireworks are readily available on the black market from illegal traders. The number of firework injuries presenting to our unit during the three week run-up to Hallowe'en October 2001 was recorded. In addition, each patient was contacted to determine how the fireworks were obtained, the average amount of money spent, and the level of adult supervision present at the time of injury. A total of 19 patients presented, 18 from the local catchment area, with a mean age of 16 yrs (range 5-46 yrs). Thirteen patients required admission. Sixteen patients sustained hand injuries including burns, and three sustained burns to other body areas. The amount of money spent varied between adults and children, the average amount among the paediatric group was Euro 2-4, but Euro 45 in the adult group. None were willing to identify the local source of their fireworks, but most fireworks originated in Northern Ireland. This small review highlights an ongoing problem in Ireland; fireworks are illegal, yet they are easily and cheaply available without quality or safety controls. Our public awareness campaign has failed to reach its target audience, and the illegal traders who sell these often inferior products are seldom charged. Children and adults will continue to sustain serious injuries as a result. PMID- 15532973 TI - Audit of the management of convulsive status epilepticus in children: the need for a uniform treatment strategy. AB - We conducted a two-year prospective audit to review the paediatric management of Convulsive Status Epilepticus (CSE) in Ireland. Our audit showed that there is considerable variability in the management of CSE in this country. In order to provide optimum care for this potentially life-threatening condition a uniform management strategy is required. We propose a protocol for the treatment of CSE, which should ensure uniform management and optimum care and also provide a template for further study and audit of this important disorder. PMID- 15532974 TI - Spontaneous keloids in siblings. AB - The authors report two rare cases of 'non-syndromic spontaneous keloids' occuring in siblings. This represents another unexplored area in the field of 'keloid challenge', warranting further research and development. PMID- 15532975 TI - Hepatitis C: lower prevalence in young persons' addiction treatment programme than in adult programmes. PMID- 15532976 TI - Travel times and geographical equity: a response to Teljeur et al. PMID- 15532977 TI - Pregnant doctors need better working conditions. PMID- 15532978 TI - Biosecurity: risk and response. PMID- 15532980 TI - Cell tropism of Salmonella enterica. AB - Salmonella serotypes are able to actively cross the intestinal epithelium, mainly but not exclusively through M cells in the follicle-associated epithelium of Peyer's patches. Once reaching the basal side of the epithelium, Salmonella serotypes are internalized by macrophages, dendritic cells, and neutrophils but are not found in fibroblasts or other mesenchymal cells. The outcome of the interaction between Salmonella serotypes and dendritic cells or neutrophils is detrimental to the pathogen. In some host species Salmonella serotypes find a safe haven from humoral defenses and neutrophils within macrophages, and replication within this niche appears to be a prerequisite for the development of a systemic infection. In other host species, macrophages can control bacterial growth and the infection remains localized to the intestine and mesenteric lymph nodes. This review summarizes our knowledge on the cellular tropism of Salmonella serotypes and the bacterial and host factors relevant for these interactions. PMID- 15532979 TI - Structural biology and structure-based inhibitor design of cholera toxin and heat labile enterotoxin. AB - Structural biology studies on cholera toxin and the closely related heat-labile enterotoxin from enterotoxigenic Escherichia coli over the past decade have shed light on the mechanism of toxin action at molecular and atomic levels. Also, components of the extracellular protein secretion apparatus that translocate the toxins across the outer membrane are being investigated. At the same time, structure-based design has led to various classes of compounds targeting different toxin sites, including highly potent multivalent inhibitors that block the toxin receptor-binding process. PMID- 15532981 TI - The porin MspA from Mycobacterium smegmatis improves growth of Mycobacterium bovis BCG. AB - Mycobacteria are characterized by an extremely thick hydrophobic cell wall restricting the permeability for small hydrophilic compounds. Recently, a new efficient porin (MspA) has been identified in Mycobacterium smegmatis, which is lacking in slow-growing mycobacteria. Since we were interested in investigating the influence of porins on growth of slow-growing Mycobacterium bovis BCG, we inserted a 3429 bp DNA fragment from M. smegmatis carrying the mspA gene in an integrative vector and transferred it into M. bovis BCG. Expression of mspA in the BCG derivative was shown by RT-PCR and Western blot. Quantification of bacterial growth on agar plates demonstrated two- to four-fold better growth of the BCG derivative with the transferred DNA compared with the reference strain. Transposon mutagenesis proved the mspA gene to be responsible for the growth enhancement. Intracellular multiplication of the BCG derivative in the mouse macrophage cell line J774 and the human pneumocyte cell line A549 was also clearly enhanced pointing to a possible role of porins in the interaction of mycobacteria with their hosts. PMID- 15532982 TI - ISSag1 in streptococcal strains of human and animal origin. AB - The chromosomal region of Streptococcus agalactiae harboring the C5a peptidase and the lmb genes displays the structure of a composite transposon. Its presence in a streptococcal strain is associated with the origin of this strain from a human host. In S. agalactiae it is flanked by two copies of the insertion element ISSag2, and the nucleotide sequence for a third IS element (ISSag1) can be found in this region. Based on amino acid sequence similarity of the deduced transposase ISSag1 belongs to the IS3 family. It is 1251 bp long and flanked by 37 bp imperfect inverted repeats. Horizontal gene transfer among different bacterial species is facilitated by mobile genetic elements. To investigate if ISSag1 homologues are also present in other streptococcal species, various species of pyogenic streptococci from animal and human origin were analyzed by Southern blot hybridization and PCR. Among the different streptococcal species, multiple copies of an ISSag1 homologue could only be detected in S. dysgalactiae subsp. dysgalactiae strains of animal origin. All of the S. agalactiae strains harbored only a single copy, that was always found in strains with the scpB-lmb composite transposon. A single copy of an ISSag1 homologue could also be detected in some of the S. pyogenes and S. dysgalactiae subsp. equisimilis strains. Nucleotide sequencing of the IS element in S. dysgalactiae subsp. dysgalactiae strains revealed several different variants. One of the variants showed the features of a regular IS3 element. The other two variants that were observed displayed a 500-bp deletion and a mosaic structure composed of ISSag1 and ISSag2 homologues. This mosaic structure suggests that recombination and horizontal gene transfer events in S. dysgalactiae strains of bovine origin could have played a role in the assembly of the scpB-lmb composite transposon structure. PMID- 15532983 TI - VTEC O157 subtypes associated with the most severe clinical symptoms in humans constitute a minor part of VTEC O157 isolates from Danish cattle. AB - The aim of this study was to compare the distribution of VTEC O157 subtypes isolated from human sporadic infections with those in the Danish bovine reservoir, and to correlate the subtypes with the severity of the clinical symptoms in humans. The study included a total of 149 Danish eae-positive VTEC O157 isolates (63 of bovine origin and 86 from human clinical cases) isolated between 1987 and 2001. All were analysed by vtx-PCR-RFLP and phage typing. The vtx-PCR-RFLP showed that isolates carrying the vtx2 gene was more than four times as prevalent among the human clinical isolates (55%) as compared to the bovine isolates (13%). Furthermore, a significant correlation between the presence of the vtx2 gene and development of haemolytic-uraemic syndrome was found. The 149 isolates encompassed 16 different phage types (PTs). The majority (87%) of the human clinical isolates were identified, as PT2, PT4, PT8 or PT14 while only 46% of the bovine isolates belonged to these PTs. PT8 and PT14 were found at similar rates among bovine (36%) and human isolates (40%). However, the predominant PTs in the human isolates, PT2 (19%) and PT4 (28%), were only identified in 2% and 8%, respectively, of the bovine isolates. All but one PT2 and PT4 isolate carried either vtx2 alone or in combination with vtx2c, whereas none of the PT8 and PT14 isolates carried vtx2. The significant overlap between vtx/phage type combinations in bovine and human clinical isolates indicate that cattle are an important reservoir for human VTEC O157 infections in Denmark. However, the vtx2 carrying isolates, causing the most severe clinical symptoms, constitute only a minor fraction of the isolates from the Danish bovine reservoir. PMID- 15532984 TI - PCR identification of the plasmid-borne enterotoxin gene (cpe) in Clostridium perfringens strains isolated from food poisoning outbreaks. AB - It is generally accepted that Clostridium perfringens strains associated with food poisoning carry their enterotoxin gene, cpe, on the chromosome, while C. perfringens strains isolated from non-food-borne diseases, such as antibiotic associated diarrhea and sporadic diarrhea, carry cpe on the plasmid. However, we recently encountered a food poisoning outbreak caused by C. perfringens bearing a plasmid cpe. We therefore investigated a total of 31 clinical and non-clinical C. perfringens strains to locate the cpe gene by PCR. The cpe of nine heat-sensitive (100 degrees C for 10min) strains isolated from three outbreaks of food poisoning were located on the plasmid, while those of six heat-resistant strains from other food poisoning outbreaks were located on the chromosome. Moreover, the cpe of 5 heat-sensitive strains isolated from healthy human feces and those of 11 heat sensitive soil strains were also located on the plasmid. These findings indicate that heat-sensitive, cpe-plasmid-borne C. perfringens strains should not be disregarded as causative agents of food poisoning. PMID- 15532985 TI - New vaccination strategies. PMID- 15532986 TI - Diphtheria-tetanus-pertussis (DTP) combination vaccines and evaluation of pertussis immune responses. AB - Diphtheria-tetanus-pertussis (DTP) combination vaccines based on inactivated whole-cell Bordetella pertussis (DTPw) or purified acellular pertussis components (DTPa) facilitate vaccine administration and will allow further co-administration such as with pneumococcal conjugates. Safety and immunogenicity studies are needed to demonstrate non-inferiority between combinations and the separate vaccines. The immunological non-inferiority is based on threshold antibody levels that represent correlates of protection. However, in case of pertussis, correlates of protection have not been defined or accepted. We describe the clinical evaluation of DTPa- and DTPw-based combinations and demonstrate their immunological non-inferiority as compared to their separately administered licensed counterparts. With respect to antibody responses against pertussis, a number of evaluations (vaccine response rates and geometric mean concentrations (GMCs) for anti-PT, anti-FHA, anti-PRN or anti-BPT; reverse cumulative distribution curves) are described. We also demonstrate that the B. pertussis mouse lung clearance model is able to predict clinical efficacy of licensed DTPa and DTPw vaccines and represents a useful tool to evaluate new combination vaccines. PMID- 15532987 TI - Pneumococcal conjugate vaccines--a European perspective. AB - Streptococcus pneumoniae is a leading cause of bacterial pneumonia, meningitis, and acute otitis media in children and adults worldwide. In the age group of < 2 years the incidence of invasive pneumococcal disease ranges from approximately 14 cases per 100,000 in Germany and the Netherlands and more than 90 per 100,000 children in Spain. The vulnerability of children to S. pneumoniae can also be demonstrated by the high rate of sequelae (> 20% in Germany) and the high mortality (7.5%) in pneumococcal meningitis. Furthermore, antibiotic resistance of S. pneumoniae is increasing in Europe, particularly in France, Spain, and Eastern European countries, whereas Germany and Northern Europe are only marginally affected. A 7-valent pneumococcal conjugate vaccine (7vPCV) that was shown to be highly efficacious in preventing invasive pneumococcal disease in infants in the USA was licensed in Europe in 2001. It is expected that broad usage of the vaccine would reduce the incidence of invasive pneumococcal disease and the levels of pneumococcal resistance significantly. Important questions have been raised regarding the effectiveness of this vaccine in high-risk populations, serotype replacement, the efficacy of this vaccine in otitis media, and the co administration of the new vaccine with other standard childhood vaccines used in various European countries. France and Spain currently have the most-wide ranging guidelines recommending pneumococcal vaccination for children. Overall, the development of pneumococcal conjugate vaccines is a significant step in the control of pneumococcal disease in children in Europe. Further progress in pneumococcal vaccine development can be expected from conjugate vaccines including more than seven serotypes (9-valent, 11-valent). PMID- 15532989 TI - Recombinant bacterial ghosts: versatile targeting vehicles and promising vaccine candidates. PMID- 15532988 TI - Genome-based vaccines. AB - Vaccination is an effective possibility to prevent many bacterial or viral infections, but for several important pathogens still no vaccines are available. The sequences of complete genomes are now decoded for an increasing number of bacterial pathogens and offer the possibility for comprehensive screenings to identify targets for vaccine development. In this article current genomic approaches to identify antigenic proteins of Neisseria meningitidis, Streptococcus pneumoniae, Staphylococcus aureus, and Chlamydia pneumoniae are summarized. PMID- 15532990 TI - "One size fits it all": translocation of foreign antigens by Yersinia type III secretion system (TTSS) leads to concomitant CD4 and CD8 T-cell priming. AB - Live replicating bacteria expressing heterologous antigens are vaccine candidates that are able to induce complex immune responses. Yersinia pseudotuberculosis employs a type III secretion system for translocation of several virulence factors directly to the cytosol of eukaryotic cells. Mice orally inoculated with an attenuated recombinant Yersinia strain translocating a chimeric Yersinia outer protein E (YopE) molecule reveal high numbers of foreign antigen-specific CD4 and CD8 T cells. Thus, cytosolic display of a single hybrid protein results in concomitant CD4 and CD8 T-cell priming. This "one-size-fits-it-all"-feature of Yersinia-translocated heterologous antigens might be advantageous to mount T cellular immune responses against complex microbes and tumors. PMID- 15532991 TI - Bacteria as DNA vaccine carriers for genetic immunization. AB - Genetic immunization with plasmid DNA vaccines has proven to be a promising tool in conferring protective immunity in various experimental animal models of infectious diseases or tumors. Recent research focuses on the use of bacteria, in particular enteroinvasive species, as effective carriers for DNA vaccines. Attenuated strains of Shigella flexneri, Salmonella spp., Yersinia enterocolitica or Listeria monocytogenes have shown to be attractive candidates to target DNA vaccines to immunological inductive sites at mucosal surfaces. This review summarizes recent progress in bacteria-mediated delivery of plasmid DNA vaccines in the field of infectious diseases and cancer. PMID- 15532992 TI - Antigen delivery by dendritic cells. AB - Dendritic cells (DC) link the innate and adaptive arms of the immune system and thus orchestrate the immune response to pathogens. A novel immune intervention strategy to control infectious diseases is based on the use of the potent immunostimulatory properties of DC for vaccination and immunotherapy. Recent advances in our understanding of DC biology and the molecular mechanisms by which DC instruct the development of an appropriate immune response to microorganisms provide means for DC-based approaches to manipulate the immune system. In experimental systems, DC vaccination has been documented to mediate protection against a wide spectrum of infectious diseases caused by viral, bacterial, parasitic and fungal pathogens. The protocols for the generation, stimulation and antigen loading of DC are being optimized, and methods for DC targeting in situ are likely to become available soon, thus paving the way for clinical applications of DC-based vaccines. PMID- 15532993 TI - CpG-DNA as immune response modifier. AB - Toll-like receptors (TLRs) have been recognized to play a fundamental role in the recognition of microbial pathogens and the activation of innate immunity. However, it has also turned out that triggering of TLRs with purified compounds or synthetic ligands represents a powerful means to modulate innate as well as adaptive immune responses. Among the substances currently under investigation as immune response modifiers CpG-DNA has gained particular interest. One reason for this is the feasibility to easily synthesize and modify immunostimulatory CpG containing oligodeoxynucleotides (CpG-ODNs). This review deals with the structural needs of CpG-ODNs. CpG-ODNs are discussed as being composed of distinct building blocks (e.g. sequence strings, backbone modifications) giving rise to a concept of structural modules within CpG-ODNs. PMID- 15532994 TI - A general elastohydrodynamic lubrication analysis of artificial hip joints employing a compliant layered socket under steady state rotation. AB - A general numerical methodology was developed in the present study to analyse the elastohydrodynamic lubrication problem of a compliant layered socket against a rigid ball under steady state rotation representing flexion and extension during walking, with particular reference to artificial hip joint replacements. The general numerical methodology consisted of using the Newton-Raphson method to solve the Reynolds equation, simultaneously with the full elasticity equation using the finite element method in combination with the fast Fourier transform technique. Two specific types of acetabular cup were considered, one with ultra high molecular weight polyethylene used in current total hip joint replacements, and one with polyurethane proposed for compliant layered 'cushion form bearings' for future developments. The film thickness and the pressure distribution for both cups were obtained under a wide range of operating conditions. The predicted central or average film thicknesses within the contact conjunction were compared with those estimated from various simplified theories available in the literature. A simple analytical methodology was consequently established to estimate the lubricating film thickness in a compliant layered socket, based on the corresponding ball-on-plane model and the consideration of the curvature effect. PMID- 15532995 TI - Contact mechanics analysis of metal-on-metal hip resurfacing prostheses. AB - Finite-element method was employed to study the contact mechanics in metal-on metal hip resurfacing prostheses, with particular reference to the effects of bone quality, the fixation condition between the acetabular cup and bone, and the clearance between the femoral head and the acetabular cup. Simple finite-element bone models were developed to simulate the contact between the articulating surfaces of the femoral head and the acetabular cup. The stresses within the bone structure were also studied. It was shown that a decrease in the clearance between the acetabular cup and femoral head had the largest effect on reducing the predicted contact-pressure distribution among all the factors considered in this study. It was found that as the clearance was reduced, the influence of the underlying materials, such as bone and cement, became increasingly important. Stress shielding was determined to occur in the bone tissue surrounding the hip resurfacing prosthesis considered in this study. However, the stress-shielding effects predicted were less than those observed in conventional total hip replacements. Both the effects of bone quality (reduction in elastic modulus) and the fixation condition between the cup and the bone were found to have a negligible effect on the predicted contact mechanics at the bearing surface. The loading was found to have a relatively small effect on the predicted maximum contact pressure at the bearing surface; this was attributed to an increase in contact area as the load was increased. PMID- 15532996 TI - Degradation of poly-L-lactide. Part 1: in vitro and in vivo physiological temperature degradation. AB - Poly-L-lactide (PLLA) is one of the most significant members of a group of polymers regarded as bioresorbable. The degradation of PLLA proceeds through hydrolysis of the ester linkage in the polymer's backbone and is influenced by the polymer's initial molecular weight and degree of crystallinity. To evaluate its degradation PLLA pellets were processed by compression moulding into tensile test specimens and by extrusion into 2 mm diameter lengths of rod, prior to being sterilized by ethylene oxide gas (EtO) and degraded in both in vitro and in vivo environments. On retrieval at predetermined time intervals, procedures were used to evaluate the material's molecular weight, crystallinity, mechanical strength, and thermal properties. Additionally, the in vivo host tissue's biological response was analysed. The results from this study suggest that in both the in vitro and in vivo environments, degradation proceeded at the same rate and followed the general sequence of aliphatic polyester degradation, ruling out enzymes contributing and accelerating the degradation rate in vivo. Additionally, the absence of cells marking an inflammatory response suggests that the PLLA rods investigated in vivo were biocompatible throughout the 44 weeks duration of the study, before any mass loss was observed. PMID- 15532997 TI - Degradation of poly-L-lactide. Part 2: increased temperature accelerated degradation. AB - Poly-L-lactide (PLLA) is one of the most significant members of a group of polymers regarded as bioresorbable. The degradation of PLLA proceeds through hydrolysis of the ester linkages in the polymer's backbone; however, the time for the complete resorption of orthopaedic devices manufactured from PLLA is known to be in excess of five years in a normal physiological environment. To evaluate the degradation of PLLA in an accelerated time period, PLLA pellets were processed by compression moulding into tensile test specimens, prior to being sterilized by ethylene oxide gas (EtO) and degraded in a phosphate-buffered solution (PBS) at both 50 degrees C and 70 degrees C. On retrieval, at predetermined time intervals, procedures were used to evaluate the material's molecular weight, crystallinity, mechanical strength, and thermal properties. The results from this study suggest that at both 50 degrees C and 70 degrees C, degradation proceeds by a very similar mechanism to that observed at 37 degrees C in vitro and in vivo. The degradation models developed also confirmed the dependence of mass loss, melting temperature, and glass transition temperature (Tg) on the polymer's molecular weight throughout degradation. Although increased temperature appears to be a suitable method for accelerating the degradation of PLLA, relative to its physiological degradation rate, concerns still remain over the validity of testing above the polymer's Tg and the significance of autocatalysis at increased temperatures. PMID- 15532998 TI - A finite element rheological model for polymethylmethacrylate flow: analysis of the cement delivery in vertebroplasty. AB - Polymethylmethacrylate (PMMA) is increasingly used in orthopaedics. Finite element (FE) modelling can play an important role in understanding the PMMA flow behaviour. However, FE models have not been used so far because conventional FE packages do not allow for the rheopectic and pseudoplastic behaviour of PMMA to be taken into consideration and because it requires multiple expertise to incorporate these behaviours into an FE package. The objectives of the present paper are to: (a) propose a rheological model that describes PMMA flow behaviour; (b) implement this model into ANSYS using FORTRAN; and (c) validate the implementation by comparing it with analytical solutions. After the validation showed good agreement, an FE model of PMMA delivery through an eight-gauge cannula was developed to examine the extra-vertebral flow conditions of vertebroplasty. The FE analysis showed a logarithmic increase of the injection pressure, where it almost doubled from 1.2 to 2.3 MPa over two minutes. This unanticipated non-linear increase is due to the highly non-uniform viscosity profile in the cannula. It can be concluded that: (a) the rheological model implemented in ANSYS can be used to analyse practical flow problems related to PMMA and (b) time and shear-rate effects of PMMA are crucial to estimate its flow behaviour accurately. PMID- 15532999 TI - Effect of joint conformity on glenoid component fixation in total shoulder arthroplasty. AB - Results of shoulder replacements are inferior and must be improved. Two of the major problems of total shoulder replacements are loosening of cemented glenoid components and wear of polyethylene inlays of uncemented, metal-backed glenoid components. The aim of this study is to investigate the influence of joint conformity on glenoid-component fixation. Keeled glenoid components, with radii of curvature of 24, 25, or 29 mm, were cemented in bone substitutes, placed in a force-controlled test set-up, articulating against a 24 mm humeral head. They were loaded by a constant joint compression force (725 +/- 10 N) and a superior subluxation force (shear force), cyclically varying between 0 and 350 +/- 1 N. After 200,000 load cycles, the upper and lower glenoid component rim displacements were measured by custom-made displacement sensors. Additionally, the shear-out strength has been measured to investigate the residual strength. The glenoid component structures with radii of curvature of 24, 25, and 29 mm showed maximum superior rim-displacements of 0.163 (SD = 0.01), 0.299 (SD = 0.0306), and 0.350 (SD = 0.0197) mm respectively, which is a significant difference (p < 0.05). The maximum shear-out strength of glenoid components with radii of curvature of 24, 25, and 29 mm was 2707 (SD = 452), 2648 (SD = 299), and 2631 (SD = 312) N respectively, which is not a significant difference (p < 0.05). However, the results indicate that a conform articulation shows smaller glenoid rim-displacements, which might be beneficial for long-term component fixation. PMID- 15533000 TI - A kinematic model of the wrist based on maximization of joint contact area. AB - The wrist is a complex joint and the factors governing its behaviour are poorly understood. A hypothesis that the movement of the carpal bones could be predicted using a minimum energy principle was tested. Carpal bones were dissected from a cadaveric forearm and their shapes were laser-digitized to obtain three dimensional computer models. A computer program was created to measure contact area between neighbouring articular surfaces and to maximize this quantity by adjusting the six degrees of freedom of the bone models. This procedure was performed for 1.0 degree increments of rotation applied to the capitate bone up to 20 degrees of ulnar and 10 degrees of radial deviation. The model correctly predicted certain aspects of the complex behaviour of the carpal bones. The results for the scaphoid in particular displayed characteristics in common with known behaviour of this bone. During 20 degrees of unlar deviation and 10 degrees of radial deviation, the bone demonstrated 11.3 degrees of extension and 9.4 degrees of flexion respectively. The novelty of the study lay in the fact that the model did not rely upon ligamentous constraints. The results are encouraging, considering the only information used by the algorithm was the shape of the articular surfaces. PMID- 15533001 TI - Temperature prediction in a finite element model for sliding contact analysis of total hip prosthesis. AB - A finite-element model for sliding contact in total hip joint prosthesis is presented in this paper. The hip prosthesis studied consists of an ultra-high molecular weight polyethylene (UHMWPE) acetabular cup articulating against cobalt chrome and alumina-ceramic femoral heads. Various aspects of prosthesis operation were analysed using the finite-element model. For example, bulk material and surface stresses were analysed under varying conditions of elastic modulus, friction coefficient, sliding speed, and radial clearance. The resulting variations of temperature were also recorded. The results obtained from the model are useful in understanding the operating conditions and the causes of wear in the hip prosthesis. PMID- 15533002 TI - Determining sources of atmospheric polychlorinated biphenyls based on their fracturing concentrations and congener compositions. AB - Air samples collected at 23 sites in Kanagawa Prefecture, Japan in 1998 and 1999 were analyzed for PCBs by a high-resolution gas chromatograph/mass spectrometer. Concentrations of PCBs ranged from 10 to 3290 pg/m3N (geometric average, 120 pg/m3N) and were mostly less than 140 pg/m3N. The toxicity equivalency quantity (TEQ) values of co-planar PCBs ranges from not detected (ND) to 0.35 pg-TEQ/m3 N (mean value, 0.00063 pg-TEQ/m3 N). Cl3-PCBs comprised the highest level in many air samples. PCB congeners in the samples collected on three consecutive days did not vary significantly, suggesting that variation of PCBs concentrations is minimal among the samples collected at same sites. On the other hand, the pattern similarities of PCBs in the samples collected at various areas were not consistent. Contamination from commercial PCBs (Kaneclor) was observed in some samples with high PCBs concentrations. The compositions of PCB congeners in the samples collected at the sites near municipal waste incinerators were similar to those of PCB congeners in the samples collected at common environments in Japan, whereas they were completely different from those in the exhaust gases collected directly from municipal waste incinerators. Consequently, contribution of exhaust gases from incinerators to PCB concentrations and/or compositions of all air samples collected in the present study was not observed. PMID- 15533003 TI - Highly toxic chlorobiphenyl and by-side impurities content and composition of technical chlorofen formulation. AB - Non- and mono-ortho CBs as well as also highly toxic by-side impurities such as chlorodibenzofurans, chlorodibenzo-p-dioxins, and chloronaphthalenes after a subsequent 2D-HPLC and HRGC-HRMS separation, detection, and identification were quantified in technical chlorobiphenyl Chlorofen formulation. Chlorofen is highly chlorinated CB mixture and its compositional profile of mono-ortho CBs is occupied by 2,3,3',4,4',5'-HxCB (no. 157) with 96.6% and followed by 2,3,3',4,4',5-HxCB (no. 156) with 3.3% and 2,3',4,4',5,5'-HxCB (no. 167) with <0.1%, while nos. 105, 114, 118, 123, and 189 remained undetected (<10 microg/g). Amongst non-ortho CBs only a trace of 3,3',4,4'-TeCB (no. 77) was found at 15 microg/g, while CBs nos. 81, 126, and 169 were absent. TCDD TEQ for mono- and non ortho CBs in Chlorofen was 2320 and 1.5 ng/g, respectively, while for CDFs and CNs were 45 and 731 ng/g, respectively, and for both types of contaminants most contributing were OcCDF and OcCN. No tetra- to OcCDD was found in Chlorofen. PMID- 15533004 TI - Simultaneous soil Cd and PCB decontamination using a surfactant/ligand solution. AB - The simultaneous desorption of Cd and PCBs, from a contaminated soil, was investigated using a surfactant/ligand solution. The test surfactants were combinations of a nonionic surfactant [polyethylene oxide (PEO) of chain length 7.5 (Triton X-114), 9.5 (Triton X-100), 30 (Triton X-305), or 40 units (Triton X 405)], with iodide (I-). Triplicate 1 g soil samples were equilibrated, during 24h, with 15 mL of surfactant/ligand solution, at 0.025 or 0.50 and 0.0, 0.168, or 0.336 mol L(-1), respectively. The supernatant fraction was then separated from the particulates by centrifugation and analyzed for Cd to determine Cd desorption efficiency. After five consecutive washings, the treated soil samples were analyzed for PCB content to determine the desorption efficiency of this class of toxicant. Desorption of Cd increased with both increasing ligand concentration and decreasing surfactant chain length and concentration. The maximum Cd desorption efficiency (61%), was obtained with Triton X-100 at 0.025 mol L(-1), in the presence of I at 0.336 mol L(-1). After 5 successive washings, virtually quantitative PCB desorption (below limits of detection for tetra-, penta-, and hexachloro biphenyl compounds) had been achieved with most of surfactant/ligand combinations. Maximum desorption efficiency was achieved using either shorter chain length surfactant (n = 7.5 or 9.5) in combination with a higher ligand concentration, or using longer chain length surfactants (n = 30 and 40) with a lower ligand concentration. Thus, heavy metals and PCBs can be desorbed simultaneously from a contaminated soil with the same washing reagents, namely a surfactant-ligand combination. The optimum surfactant-ligand combination for the simultaneous desorption of both heavy metals and PCBs was a short chain length (n = 7.5 or 9.5) surfactant at low concentration admixed with 0.336 mol L( 1) I-. PMID- 15533005 TI - The concentration of mild-extracted polycyclic aromatic hydrocarbons in sewage sludges. AB - The present study evaluates the content of the mild-solvent extracted fraction of the polycyclic aromatic hydrocarbons. Ten compounds from the US EPA list (phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chryzene, benzo[b]fluoranthene, benzo[a]pyrene, benzo[ghi]perylene oraz indeno[1,2,3 cd]pyrene) were chosen. The compounds were extracted with n-butanol from 11 sewage sludges that differed both in the total PAH content and in their physicochemical properties. On the basis of the results obtained, the influence of the properties of the PAH and some properties of selected sludge on the content of the mild-solvent extracted fractions of these compounds was determined. The content of the fraction extracted with n-butanol within the total of 10 PAHs ranged from 12.5 to 83.2% (mean 40.1%) depending on the type of sludge. Similarly, as in the case of the total of the 10 PAHs, significant differences in the content of PAHs extracted with n-butanol were also noted for the individual compounds studied. In the case of most individual PAHs their mean share did not differ statistically in relation to the PAH type and was close to 40%. An evaluation of the relation between the share of the fraction extracted with n-butanol from the different types of sludge and the properties of the PAHs showed that statistically, there existed significant (P < 0.05) correlations between log Koc (in the case of one sewage sludge) and between the nitrogen content, the ratio of C/N, cations Mg2+ and K+ (in the case of a few PAHs). A clear and significant relation was found between the content of and the share of the fraction of n-butanol extracted and the PAH fraction present in the sewage sludge pore water (determined on the basis of an equilibrium partition model). PMID- 15533006 TI - Assessment of androgenicity in leachates from Swedish landfills and treatments for its elimination. AB - An in vitro recombinant yeast strain, transfected with the human androgen receptor was used to assess androgenic hormone disrupting potencies in leachates from Swedish landfills. It was shown that components in extracts of these affected the androgenic receptor and promoted a response in the beta galactosidase marker system. Levels were within the range of those determined for domestic sewage effluents but lower than the highest levels found in an industrial effluent. These leachates finally enter receiving waters with or without wastewater treatment. Evidence was found for transformation during some of the wastewater treatments. PMID- 15533007 TI - Boron and selenium contamination in south Texas groundwater. AB - Boron and selenium concentrations from 112 water wells in an irrigated agricultural region of south Texas were compiled, mapped, and statistically analyzed. Wells in the study area produce water from the Gulf Coast Aquifer System, comprising coastward sloping beds of clay, silt, sand, and gravel. Nearly 84 percent of boron observations exceeded the 600 ug/L advisory level for drinking water, 70% exceeded the 1250 ug/L level for sensitive crops, and 24% exceeded the 3750 ug/L level for tolerant crops. Additionally, 21% of selenium observations exceeded the 20 ug/L advisory level for irrigation water, and five percent surpassed the 50 ug/L standard for drinking water. Many wells with high boron concentrations also had high selenium concentrations, and several clusters of high concentrations were in irrigated parts of the study area. However, there was no association between selenium and well depth, and a direct rather than inverse association between boron and well depth. Progressively brackish water in downdip reaches of the aquifer largely controls observed boron patterns. Both irrigation practices and prevailing groundwater chemistry significantly influence selenium concentrations in the study area. PMID- 15533008 TI - Investigation of the formation of chlorination by-products in water rich in bromide and organic matter content. AB - Bench-scale chlorination experiments with river water rich in bromide and organic matter content were performed in order to investigate the behavior and speciation of the chlorination by-products, especially the brominated ones, which are considered more harmful to human health than their chlorinated analogues. The analysis of the compounds was performed by means of gas chromatography. Statistical treatment of the results with Multifactor Analysis of Variance technique revealed the trends of each compound as function of pH, chlorine dose, reaction time and temperature. Brominated species of chlorination by-products predominated, due to the presence of bromide in the river water leading to preferable bromination than chlorination of the organic matter. Bromide incorporation factors were calculated for trihalomethanes and haloacetic acids, the major categories of compounds formed. While bromide incorporation factors decreased with increasing chlorine dose, the opposite was true for low range of chlorine dose, a fact that needs to be considered during treatment of waters with similar properties. Bromide incorporation factors decreased with increasing pH, especially for haloacetic acids. While the organic matter content can be sufficiently removed during water treatment, bromide ion still remains in the treated water. The results of the present study indicate that chlorination conditions should be optimized in order to minimize the incorporation of bromide into the chlorination by-products formed, which would increase their toxic effects on the water consumers. PMID- 15533009 TI - Organo-silicate nanocomposites for the removal of chlorinated phenols from aqueous media: kinetics and environmental stability. AB - Organosilicate nanocomposite hexagonal mesostructure (NHMS) was synthesized from dodecylamine and tetraethyl orthosilicate (TEOS). The nanocomposite (NMCM) of numbers 41, 48, and 50 were synthesized from cetyltrimethylammonium bromide (CTABr) and (TEOS). The rates, affinity, and stability of these synthetic nanocomposite materials to remove and retain chlorinated phenols from aqueous solution were investigated; all materials have the ability for sorption and retention of 2,4-dichlophenol (2,4-DCP). Batch absorption kinetics indicates that NHMS, NMCM-41, NMCM-48, and NMCM-50 are 1st order reactions, with rate constants of 0.412, 0.296, 0.112, and 0.0161 hr(-1), respectively. Average percent 2,4-DCP removal for NHMS, NMCM-41, NMCM-48, and NMCM-50 was 92, 98, 90, and 52%, respectively. Isothermic measurements fit Freundlich and Langmuir models. NHMS and NMCM-41 best fit Langmuir model. Stability of the adsorbents in 0.10 M CaCl2 for 56 days was in the order: NMCM-41 > NHMS > NMCM-50 > NMCM-48. PMID- 15533010 TI - Photocatalytic degradation of nonionic surfactants with immobilized TiO2 in an airlift reactor. AB - The photocatalytic degradation of a nonionic surfactant, Sanonic SS-90 (polyoxyethylene alkyl ether) which is one of polyoxyethylene nonionic surfactants, in water has been investigated using immobilized titanium dioxide (TiO2) photocatalyst. An external UV irradiation type airlift reactor was used as a photoreactor. The effects of initial nonionic surfactant concentration, intensity of UV light, and concentrations of TiO2 photocatalyst on degradation rate of Sanonic SS-90 were examined. The photocatalytic decomposition of Sanonic SS-90 was assumed to be approximately described by the Langmuir-Hinshelwood kinetics. The rate constant of photocatalytic degradation was found to be proportional to TiO2 surface area and the square root of UV light intensity and independent on the initial concentration of Sanonic SS-90. The evaluated average UV light intensity in the airlift photoreactor was used to determine the rate constants in the Langmuir-Hinshelwood kinetics. It was found that the kinetic model for the degradation by immobilized TiO2 photocatalyst can simulate the experimental results reasonably. PMID- 15533011 TI - Heavy metals in the products of deinking flotation of digital offset prints. AB - This study was undertaken to investigate the suitability of applying the conventional method of chemical deinking flotation of digital offset prints and ecological implications of the disposal of digital offset prints, with special emphasis on the content of heavy metal cations. The EDXRF method was used to determine the concentrations of Pb, Zr, Sr, As, Co, Zn, Cu, Ni, Fe, Mn, Cr, V, Ti, Ca, and K cations in handsheet ashes after flotation as well as in ashes of foams separated from cellulose suspension in the flotation phase. The same method was applied to determine the concentrations of Pb, Cr, Mn, Fe, Ni, Cu, Zn, and Co in process water filtrates obtained from the cellulose suspension after flotation and in foam filtrates. Centrifuging and coagulation previously treated flotation process water, while foam filtrates were centrifuged. Concentrations of Pb, As, Sr, Zr, K, Ca, Ti, V, Cr, Mn, Fe, Cu, and Zn were determined in the sediments obtained by centrifuging process water. The trials were done with unprinted substrates without deinking chemicals, with unprinted substrates in the presence of deinking chemicals and with printed substrates in the presence of deinking chemicals. Cation analysis revealed that deinking chemicals facilitate release of cations from the substrate and their transition into process water. Concentrations of most cations in the flotation process water, in foam water as well as in the sediment of the process water of printed substrate flotation deinking were lower than those obtained by flotation of unprinted substrate suspension. Polymer particles of ElectroInk incompletely separated from cellulose fibers prevent release of cations from cellulose fibers into process water. Deinking chemicals do not affect release of cations from ElectroInk particles, so the substrate is the main source of increased concentration of cations in the water of digital offset print deinking. The major part of cations in process water is bound to the colloid particles of the filler (CaCO3) released by substrate disintegration and to particles of fine fibers. PMID- 15533012 TI - Characteristics of p-hydroxybenzoic acid oxidation using Fenton's reagent. AB - The objective of this work was to investigate the oxidation of p-hydroxybenzoic acid by the Fenton's reagent. Batch experiments were carried out to investigate the influence of the most important process variables: hydrogen peroxide and ferrous salt concentrations, pH, and temperature. The optimal results obtained indicate that p-hydroxybenzoic acid can be effectively degraded using Fenton's oxidation within 10-min reaction time by using a molar ratio H2O2: p hydroxybenzoic acid of 4:1 and H2O2:Fe2+ of 15:1, at 30 degrees C and pH=3.0. The behavior of ORP and pH along the reaction time for different values of R (molar ratio H2O2: p-hydroxybenzoic acid) was also discussed. A pseudo-first order model was applied to describe the oxidation kinetics of p-hydroxybenzoic acid by Fenton's reagent. PMID- 15533013 TI - Aqueous ozone decomposition onto a Co2O3-alumina supported catalyst. AB - The aqueous ozone decomposition in the presence of a Co2O3-Alumina catalyst was investigated. Activity and estability assays were conducted by reusing the same catalyst in consecutive runs. The catalyst was shown to significantly increase the ozone abatement rate without loss in activity after five consecutive experiments. The process can be acceptably simulated by a double homogeneous heterogeneous decomposition mechanism. An increase in the working temperature resulted in an opposite effect by increasing the extension of the homogeneous decomposition and lowering the extension of the heterogeneous decomposition. Similarly, different trends were observed by adding two distinct free radical scavengers (terc-butyl alcohol and carbonates). PMID- 15533014 TI - Characteristics of cadmium uptake and accumulation by two contrasting ecotypes of Sedum alfredii Hance. AB - The mined ecotype of Sedum alfredii Hance has been identified to be a zinc (Zn) hyperaccumulator native to China. In the present article, the characteristics of cadmium (Cd) uptake and accumulation were compared with hydroponic experiments between the mined and the nonmined ecotypes of Sedum alfredii Hance. The results indicate that the plants of the mined ecotype (ME) have higher tolerance of Cd than the plants of the nonmined ecotypes (NME) in terms of dry matter yield. The thresholds of external Cd levels for the reduction of plant growth were 100 micromolL(-1) for the NME and 400 micromolL(-1) for the ME, respectively. Kinetic study showed that the rates of Cd influx into roots (IR) and transport to shoots (TR) were higher in the ME than in the NME, with 5-fold higher for the maximum IR (Imax) and 13-fold higher for the maximum TR (Tmax) in the NME, respectively. Cadmium concentrations increased with increasing external Cd supply levels. Root Cd concentrations in the NME were higher than that in the ME, with a maximum being 5646 mg kg(-1) for the NME and 2889 mg kg(-1) for the ME at 1000 micromolL( 1) Cd. On the contrary, shoot Cd concentrations of the NME were far lower than that of the ME. Maximum shoot Cd concentrations were 533 mg kg(-1) in leaves and 935 mg kg(-1) in stems at 1000 micromol L(-1) Cd for NME, whereas, 4933 and 3874 mg kg(-1) at 400 micromol L(-1) Cd for the ME, respectively. Meanwhile, Cd concentrations in the shoots of both the NME and ME increased with advancing Cd treatment time. At 100 micromolL(-1) Cd, concentrations of Cd in leaves and stems of the NME sharply increased within initial 8 and 12 days, and those in the ME increased dramatically until D20 and D16, respectively. However, leaf and stem Cd concentrations reached their maximum values on D4 for the NME and D8 for the ME, respectively, when the plants were exposed to 1 micromol L(-1) Cd. Cadmium accumulation by plant shoots was obvious higher in the ME than in the NME at varied Cd supply levels or Cd treatment time. The maximum Cd taken up by the shoots was 1032 microg plant(-1) in concentration-dependent uptake, and 1699 microg plant(-1) in time-course uptake for the ME, with 15-fold and 18-fold higher than those for the NME, respectively. The ratios of shoot/root of Cd ranged from 12 to 39, varying with Cd supply levels, and from 13 to 24 in the varied treatment times for the ME, more than 10 times greater than those for the NME. In addition, Cd distribution in leaves, stems and roots of ME was greatly different from those of NME. The percentage of Cd distribution in shoots was more than 79 at the varied Cd supply levels, or 83 in the varied treatment time for ME, both higher than that for NME. It could be concluded that the mined ecotype of Sedum alfredii Hance has a greater ability to tolerate, transport, and accumulate Cd, as compared with the nonmined ecotype. PMID- 15533015 TI - Effect of wastewater composition on methanogenic activity in an anaerobic reactor. AB - In this study, effects of a chemical synthesis-based pharmaceutical wastewater on activity of acetoclastic methanogens within a lab-scale anaerobic completely stirred tank reactor (CSTR) were investigated. The CSTR was initially fed with glucose and loaded up to an organic loading rate of 6 kg COD m(-3) d(-1) at which 92% soluble COD removal efficiency was achieved. Acetoclastic methanogenic activity was found to be 336 mL CH4 gTVS(-1) d(-1). The CSTR was then, fed with several dilutions of preaerated wastewater with glucose at 10% wtv(-1), 30% wtv( 1), and 70% wtv(-1) ratios and followed by 100% wtv(-1) preaerated wastewater. In the latter, 71% soluble COD removal efficiency was obtained and a maximum acetoclastic methanogenic activity was 166 mL CH4 gTVS(-1) d(-1). Finally, raw wastewater diluted with preaerated wastewater was fed into the CSTR in increasing ratios of 10-60% wtv(-1). Total failure of anaerobic reactor was observed at 60% wtv(-1) raw wastewater fed. The study was, therefore, discontinued due to poor acetoclastic methanogenic activity of reactor sludge which was totally distorted. PMID- 15533016 TI - Effect of aeration on the performance of a simulated landfilling reactor stabilizing municipal solid wastes. AB - In this study, the effects of intermittent aeration on the anaerobic treatment of domestic solid waste and leachate characteristics were investigated in three simulated landfill anaerobic bioreactors. All of the reactors were operated with leachate recirculation and recirculation rate was 300 mL/d. All the reactors were loaded with solid waste having different operational mode. The first reactor was operated with recirculation (control-no aeration). The second reactor was aerated three days in a week during 1 h (Run 1) and the last reactor was aerated one day in a week during 1 h (Run 2). pH, chemical oxygen demand (COD), volatile fatty acids (VFA), ammonium nitrogen (NH4-N), nitrate nitrogen (NO3-N) concentrations, total, and methane gas productions in the leachate samples were regularly monitored. 96, 85, and 97% COD and 86, 77, and 93% VFA removals were obtained, in the leachate samples of the control, R1 and R2 reactors, respectively, after 79 days of anaerobic incubation. The TN, TP and NH4-N concentrations in MSW reduced to 4300, 1100, and 160 mg/g from 11,100, 2450, and 630 mg/g in R1 reactor by day 79. The values of pH were 6.98, 6.76, and 7.98, respectively, after anaerobic incubation, respectively in the aforementioned reactors. It was observed that the aeration decreased the methane percentage and the quantity in the simulated bioreactor. The maximum cumulative methane gas production was recorded as 11.2, 0.9, and 3.6 L in control, R1 and R2 reactors, respectively, at the end of 79 days. A BOD5/COD ratio of 0.23 achieved in the R2 reactor indicated the better MSW stabilization resulting in a high rate than that of R1 and control reactors. It was observed that the aerated reactor one day in a week during one hour (R2) reduced the COD and VFA concentrations in leachate samples. The aerated reactor three days in a week during 1 h (R1) reduced the waste quantity, the organic content of the solid waste. The final leachate quantities of aerated runs were lower than that of control reactor operated under strictly anaerobic conditions. PMID- 15533017 TI - Microbial recovery of copper from printed circuit boards of waste computer by Acidithiobacillus ferrooxidans. AB - The bioleaching of copper contained in the printed circuit boards (PCB) of waste computers by A. ferrooxidans was studied. The Fe oxidation rates by A. ferrooxidans in the 9K medium supplemented with the leachate of PCB (0.15-0.13 g L(-1) d(-1)) were similar to that in the 9K medium without the leachate (0.15 g L(-1) d(-1)). This finding suggests that the leachate of PCB did not seriously affect the bioleaching process by this bacterium. The amount of copper leached from PCB shreds increased with the addition of ferrous ion and reached up to 5190 mg L(-1) when the initial concentration of Fe2+ ion was 7 g L(-1). As the microbial leaching progressed, pale brown precipitate was observed to form in the solution. Based on the total amount of copper, both in solution and precipitate, the optimal addition of ferrous ion for the leaching of copper was around 7 g L( 1). When citric acid was not added, only about 37 wt% of the total leached copper remained dissolved; however, the amount of dissolved copper increased to greater than 80 wt% in the presence of citric acid. This fact indicates that the addition of a complexing agent (citric acid) to the bioleaching solution can raise the solubility of the leached metal ions. PMID- 15533018 TI - Effect of combined nutrients on biosurfactant produced by Pseudomonas putida. AB - This work investigated biosurfactant production by Pseudomonas putida in combined C/P, C/Ninorganic, C/Fe, C/Mg nutrient ratios and peptone concentration. Analysis of the 2(5-1) fractional factorial experimental design showed that only the C/Fe ratio had a significant (p<0.02) effect on biosurfactant production. The highest amount of biosurfactant was obtained at low C/Fe ratios, but net surface tension did not show significant differences. In addition, low amounts of peptone and the C/P-C/Mg nutrient ratios interaction significantly (p < 0.05) enhanced the biomass produced by P. putida. Analysis of biosurfactant by gas chromatography (GC) showed that the hydrophilic fraction was composed by rhamnose and the hydrophobic fraction, mainly by palmitic (C16), stearic (C18:0), oleic (C18:1) and linoleic (C18:2) fatty acids. PMID- 15533019 TI - Effects of nitrogen forms on the production of cyanobacterial toxin microcystin IR by an isolated Microcystis aeruginosa. AB - A cyanobacterial strain, which produced high content of microcystin-LR (MC-LR) but no microcystin-RR (MC-RR), was isolated from the hypertrophic Dianchi Lake in China and identified as Microcystis aeruginosa DC-1. Effects of nitrogen containing chemicals and trace elements on the growth and the production of MC-LR by this strain were studied. In the presence of bicine, compared with urea and ammonium, nitrate greatly promoted the growth and the production of MC-LR. However, leucine and arginine, which were the constitutional components in the molecular structure of MC-LR or RR, inhibited the production of MC-LR. Iron and silicon up to 10 mg/L had little effects on the growth of M. aeruginosa DC-1, but the production of MC-LR was apparently enhanced. Under all conditions studied here, only MC-LR but no RR was detected within the cells of M. aeruginosa DC-1. Thus, chemical forms of nitrogen, rather than the usually concerned the total nitrogen, and trace elements played important roles in the production of MC toxins during cyanobacterial blooms. PMID- 15533020 TI - Concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation to the occurrence of pathological spermatozoa. AB - In this study the concentration of copper, iron, zinc, cadmium, lead, and nickel in bull and ram semen and relation of these metals to spermatozoa morphology was investigated. Analysis by atomic absorption spectrophotometry showed that copper concentration was significantly higher (p<0.0001) in ram semen in comparison with bull semen. The zinc concentration was higher in bull semen in comparison with ram semen. The iron and cadmium concentrations in the semen were similar. Higher concentration of lead was found in ram semen. Higher levels of nickel were found in ram semen in comparison with bulls. In bull semen 11.79+/-4.88% of pathological spermatozoa was found. Higher occurrence of pathological spermatozoa was in ram semen (17.17+/-3.76) in comparison with the semen of bulls. Separated tail, tail torso, and knob twisted tail were the most frequent forms of pathological spermatozoa in both species. Correlation analysis in bulls showed high positive relation between iron and zinc (r = 0.72), nickel and separated tail (r = 0.76), separated tail and tail torso (r = 0.71), tail torso and total number of pathological spermatozoa (r=0.72), and between tail ball and total number of pathological spermatozoa (r = 0.78). In rams high positive correlation between cadmium and lead (r=0.98), nickel and separated tail (r=0.77), separated tail and total number of pathological spermatozoa (r=0.69), knob twisted tail and retention of cytoplasmic drop (r=0.78), and between knob twisted tail and other pathological spermatozoa (r = 0.71) was found. High negative correlation in ram semen was observed between copper and nickel (r=0.71), copper and separated tail (r=0.70), and between iron and tail torso (r=0.67). The results suggest that the studied metals have a direct effect on spermatozoa quality. PMID- 15533021 TI - Utilization of steel slag as an adsorbent of ionic lead in wastewater. AB - The feasibility of using slag, a waste from steel-making industry, as an adsorbent for ionic lead in wastewater was studied. Kinetic and equilibrium aspects of Pb2+ adsorption on the slag were investigated along with the effects of temperature and pH. Additionally, adsorption change at different ionic strength and desorption characteristics were also addressed. We identified that the adsorption kinetics of Pb2+ on the slag follows a first order reaction that can be modeled by Freundlich adsorption isotherm. It was also found that equilibrium adsorption of Pb2+ decreases with pH, which may be associated with the change of electrokinetic potential of slag as a function of pH. As temperature increases the adsorption capacity rises, and the data can be best fitted using van't Hoff equation. Ionic strength was found to negatively affect the adsorbability of Pb2+. Most of the adsorbed Pb2+ could be desorbed rapidly by a complexing agent, such as EDTA, which may suggest the feasibility of recycling of slag as an adsorbent for Pb2+. PMID- 15533022 TI - Integration of advanced oxidation technologies and biological processes: recent developments, trends, and advances. AB - The greatest challenge of today's wastewater treatment technology is to optimize the use of biological and chemical wastewater treatment processes. The choice of the process and/or integration of the processes depend strongly on the wastewater characteristics, concentrations, and the desired efficiencies. It has been observed by many investigators that the coupling of a bioreactor and advanced oxidation processes (AOPs) could reduce the final concentrations of the effluent to the desired values. However, optimizing the total cost of the treatment is a challenge, as AOPs are much more expensive than biological processes alone. Therefore, an appropriate design should not only consider the ability of this coupling to reduce the concentration of organic pollutants, but also try to obtain the desired results in a cost effective process. To consider the total cost of the treatment, the residence time in biological and photochemical reactors, the kinetic rates, and the capital and operating costs of the reactors play significant roles. In this study, recent developments and trends (1996-2003) on the integration of photochemical and biological processes for the degradation of problematic pollutants in wastewater have been reviewed. The conditions to get the optimum results from this integration have also been considered. In most of the studies, it has been shown that the integrated processes were more efficient than individual processes. However, slight changes in the configuration of the reactors, temperature, pH, treatment time, concentration of the oxidants, and microorganism's colonies could lead to a great deviation in results. It has also been demonstrated that the treatment cost in both reactors is a function of time, which changes by the flow rate. The minimum cost in the coupling of the processes cannot be achieved unless considering the best treatment time in chemical and biological reactors individually. PMID- 15533023 TI - An integrated subsurface modeling and risk assessment approach for managing the petroleum-contaminated sites. AB - Soil and groundwater contamination can lead to a variety of impacts and risks to the communities. Identifications of management schemes with sound environmental and socio-economic efficiencies is desired. In fact, before any decisions regarding site remediation actions can be made, three major questions may have to be answered. They include "What happened underground, and what will happen in the future under the given remediation scenarios?," "Are there specific risks on the surrounding community?" and "What remediation alternatives are suitable for the site?" In this study, an integrated subsurface modeling and risk assessment method for petroleum-contaminated site management is proposed. It incorporates multi-phase flow multi-component transport modeling and ELCR-based human health risk assessment into a general framework. The proposed method is applied to a case study within a western Canada context for identifying effective management schemes with improved environmental and socio-economic efficiencies. Given conditions at the study site, six remediation alternatives based on combinations of several technologies are recommended, with the provision of analyses for equipment/manpower requirements, system designs, operations, efficiencies, and costs. These alternatives can be categorized into two groups: hybrid ex situ and in situ remediation approaches, and integrated in situ remediation approaches. This study is a new attempt that integrates issues of subsurface-contamination simulation, risk assessment, and site remediation for a real-world problem within a general research framework. The research outputs are directly useful for the industry to gain insight of the site and to make decisions of the relevant remediation actions. PMID- 15533024 TI - Position paper: whole bowel irrigation. AB - Whole bowel irrigation (WBI) should not be used routinely in the management of the poisoned patient. Although some volunteer studies have shown substantial decreases in the bioavailability of ingested drugs, no controlled clinical trials have been performed and there is no conclusive evidence that WBI improves the outcome of the poisoned patient. Based on volunteer studies, WBI should be considered for potentially toxic ingestions of sustained-release or enteric coated drugs particularly for those patients presenting greater than two hours after drug ingestion. WBI should be considered for patients who have ingested substantial amounts of iron as the morbidity is high and there is a lack of other options for gastrointestinal decontamination. The use of WBI for the removal of ingested packets of illicit drugs is also a potential indication. WBI is contraindicated in patients with bowel obstruction, perforation, ileus, and in patients with hemodynamic instability or compromised unprotected airways. WBI should be used cautiously in debilitated patients or in patients with medical conditions that may be further compromised by its use. The concurrent administration of activated charcoal and WBI may decrease the effectiveness of the charcoal. The clinical relevance of this interaction is uncertain. A review of the literature since the preparation of the 1997 Whole Bowel Irrigation Position Statement revealed no new evidence that would require a revision of the conclusions of the Statement. PMID- 15533025 TI - Carbon monoxide and cyanide poisoning in fire related deaths in Victoria, Australia. AB - OBJECTIVE: This study was undertaken to examine the association of hydrogen cyanide and carboxyhaemoglobin in victims of fire related deaths in Australia. The secondary aim was to document demographic data about Australian fire related deaths. METHODS: An observational retrospective study was undertaken of autopsy reports from the Victorian Institute of Forensic Medicine. Reports of fire related deaths were electronically searched using the terms burns, "smoke" or "fire" as a cause of death in the calender years 1992 to 1998. Data on the circumstances of the fire and results of toxicological screening were obtained on 178 persons. Additional whole blood cyanide levels were determined if blood samples were available in storage. Demographics of the victims were analysed, as well as the relationship between carboxyhaemoglobin and whole blood cyanide levels. RESULTS: Most (82%) of the victims died at the scene, whilst 32 victims died after a period of hospitalisation (hours to weeks). Suicide as a result of self-immolation was the reported cause of death in 32 cases. Most of the fires were in houses (114) and cars (29). The blood ethanol level was zero in 112 cases; the remaining cases (53) had a mean level of 0.17%. Other central nervous system (CNS) depressants were recorded in 49 of the 134 cases that received a complete toxicological screen. Carboxyhaemoglobin levels were measured in only 154 of 178 cases. The carboxyhaemoglobin level was zero in 43 cases. The remaining cases (111) had a mean level of 40%; with 44 cases having a level greater than 50%, a level considered to be potentially lethal. Whole blood hydrogen cyanide levels were measured in only 138 of 178 cases. The hydrogen cyanide level was zero in 52 cases. The remaining cases (86) had a mean level of 1.65 mg/L; with 11 cases having a level greater than 3.0 mg/L (potentially fatal). Blood ethanol levels were significantly correlated with both carboxyhaemoglobin (R = 0.22, P < 0.01) and cyanide (R = 0.36, P < 0.001). In addition, a significant correlation (r = 0.34) between carboxyhaemoglobin and hydrogen cyanide levels was noted. CONCLUSIONS: This study showed a correlation between elevated blood ethanol and whole blood cyanide levels (r = 0.36, p < 0.001) and between elevated carboxyhaemoglobin and hydrogen cyanide levels (r = 0.34). Although the mean cyanide level was 1.3 mg/L (above the level some consider potentially toxic) in those cases with a carboxyhaemoglobin level of greater than 10%, there is insufficient data to permit recommendations for clinical care. Further studies are required on those victims that reach hospital alive. PMID- 15533026 TI - Speed of initial atropinisation in significant organophosphorus pesticide poisoning--a systematic comparison of recommended regimens. AB - OBJECTIVE: Early deaths from organophosphorus (OP) pesticide self-poisoning result from respiratory failure and cardiovascular collapse. Therapy requires the urgent use of atropine to reverse cholinergic excess, thereby improving respiratory function, heart rate, and blood pressure. We aimed to assess variation in textbook recommendations for early atropinisation and to see whether this variation affected time to stabilisation using model data from 22 severely poisoned patients seen in a Sri Lankan clinical trial. METHODS: We extracted prospectively recorded data on atropine requirements for 22 OP poisoned patients who required intubation but survived to discharge. We did a systematic search for textbook recommendations for initial atropinisation regimens. These regimens were then applied to data from the Sri Lankan patients. RESULTS: The patients required a mean of 23.4 mg (standard deviation 22.0, range 1-75 mg) atropine to clear the lungs, raise the pulse above 80 bpm, and restore systolic blood pressure to more than 80 mmHg. Textbook recommendations varied markedly--atropinisation of an average patient, requiring the mean dose of 23.4 mg, would have taken 8 to 1380 mins; atropinisation of a very ill patient, requiring 75 mg, would have taken 25 to 4440 mins. Atropinisation was attained most rapidly with a regimen of increasing bolus doses after failure to respond to the previous bolus. CONCLUSIONS: There is great variation in recommendations for atropinisation, with some regimens taking hours and even days to stabilise a patient. The guidelines are very flexible--possibly appropriate for experienced emergency physicians or clinical toxicologists, but completely inappropriate for the inexperienced junior doctors who see most cases worldwide. We recommend that a consensus guideline be developed by appropriate organisations to bring order to this important part of OP therapy, while acknowledging the paucity of data to drive the guidelines. PMID- 15533027 TI - A meta-analysis of prognostic indicators to predict seizures, arrhythmias or death after tricyclic antidepressant overdose. AB - OBJECTIVES: To systematically review and summarize studies on the accuracy of ECG and tricyclic antidepressant (TCA) concentration as prognostic indicators of the risk of seizures, ventricular arrhythmia (VA) or death in patients with TCA overdose. METHODS: Articles were identified with MedLine and Cochrane register of controlled clinical trials searches and review of medical toxicology textbooks. Quality of the included studies was assessed. Pooled estimates of sensitivity, specificity, likelihood ratios and Summary Receiver Operating Characteristics (SROC) curves were generated. RESULTS: A total of 18 studies were included in the analysis. The pooled sensitivity (Se) and specificity (Sp) of the QRS for predicting seizures were 0.69 [95% CI 0.57-0.78] and 0.69 [95% CI 0.58-0.78] as compared to 0.75 [95% CI 0.61-0.85] and 0.72 [95% CI 0.61-0.81] for the TCA concentration. The Se and Sp of the QRS to predict VA were 0.79 [95% CI 0.58 0.91] and 0.46 [95% CI 0.35-0.59] compared to 0.78 [95% CI 0.56-0.90] and 0.57 [95% CI 0.46-0.67] for the TCA concentration. The Se and Sp of the QRS to predict death were 0.81 [95% CI 0.54-0.94] and 0.62 [95% CI 0.55-0.68] compared to 0.76 [95% CI 0.49-0.91] and 0.60 [95% CI 0.47-0.72] for the TCA concentration. Very few studies evaluated the accuracy of QTc, T 40 ms axis and the R/S ratio. CONCLUSIONS: Overall, the studies suggested that the ECG and TCA concentration have similar but relatively poor performance for predicting complications, such as seizures, VA or death, associated with TCA overdose. PMID- 15533028 TI - Survival after a lethal dose of arsenic trioxide. AB - A case of a 27-year-old woman who ingested 9000 mg arsenic trioxide (As2O3) is reported. Classical symptoms of an acute arsenicum (As) poisoning such as gastrointestinal cramps, vomiting, diarrhea, ECG changes and disturbed liver function tests were observed. The absorption of the ingested As was minimalized by a continuous gastric irrigation with highly concentrated NaHCO3 and intestinal cleansing with NaHCO3 and polyethyleneglycol was performed. Forced diuresis, BAL (2,3-dimercaptopropanol) and DMSA (meso-2,3-dimercaptosuccinic acid) were started and therapy to enhance the formation of methylated As derivatives, which are potentially less toxic and which can be excreted more easily, was then administered. The patient survived this massive dose of ingested inorganic As with only polyneuropathy one year later. PMID- 15533029 TI - Rapid cyanide detection using the Cyantesmo kit. AB - BACKGROUND: Sources of cyanide exposure are many, including combustion of plastic and vinyl, such as in a house fire, laboratory or industrial exposures including exposure in the electroplating industry both of printed circuit boards and in jewelry work. Rapid and definitive diagnosis of cyanide poisoning is unavailable in the emergency department setting. It is desirable to make a definitive diagnosis in order to prevent potential complications of empiric treatment of presumptive cyanide poisoning from the cyanide antidote kit currently approved by the US Food and Drug Administration (FDA). We investigated a technique to detect cyanide currently utilized by water treatment facilities to determine if it can be applied to rapidly detect concentrations of cyanide in the clinically important range. METHODS: Varying standardized dilutions of KCN ranging from 0.25 microg/mL to 30 microg/mL were acidified with a drop of sulphuric acid in a closed system under a ventilation hood. Cyantesmo test strips were placed into the test tubes above the fluid level where liberated HCN gas interacted with the test strip to effect a color change. Color changes were compared to negative controls and to each other. RESULTS: The test strips demonstrated an incrementally increasing deep blue color change over a progressively longer portion of the test strip in less than 5 minutes for each concentration of KCN including 1, 3, 10, and 30 microg/mL. The concentrations of 0.25, 0.5, and 0.75 required more than 2 hours to begin demonstration of any color change. CONCLUSION: The Cyantesmo test strips accurately and rapidly detected, in a semi quantifiable manner, concentrations of CN greater than 1 microg/mL contained in each test sample. Future work to validate this test in blood and in clinical specimens is planned. PMID- 15533030 TI - Diagnostic accuracy of urinary amanitin in suspected mushroom poisoning: a pilot study. AB - BACKGROUND: Amatoxin-containing species are responsible for the most severe cases of mushroom poisoning, with high mortality rate. Therefore, this poisoning should be ruled out in all patients presenting gastrointestinal symptoms after wild mushroom ingestion. OBJECTIVE: To determine sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic efficacy (DE) of urinary amanitin analysis in cases of suspected mushroom poisoning. METHODS: All cases of mushroom ingestion referred to a Poison Center during a one-month period were analyzed. Amanitin measurements were performed by ELISA method (functional least detectable dose 1.5 ng/ml; cut-off value not clearly established). Gastrointestinal symptoms latency and initial clinical assessment were considered alternative diagnostic tools. Definitive diagnosis was used as the reference standard. RESULTS: Among 61 patients included in the study, amatoxin poisoning was diagnosed in 10 cases. Urine samples were collected 5.5 to 92 hours after mushroom ingestion. Urinary amanitin DE was 91.8%, 93.4%, and 80.3%, based on the cut-off value considered (1.5, 5.0, and 10.0 ng/ml, respectively). Symptoms latency longer than 6 hours and initial clinical assessment DE were 70.5% and 67.2%, respectively. To identify amatoxin poisoning, initial clinical assessment resulted more sensitive and urinary amanitin analysis more specific. CONCLUSIONS: Urinary amanitin analysis is a valuable diagnostic tool and may significantly contribute to the management of suspected mushroom poisoning. At present, the best diagnostic accuracy can be obtained taking advantage of both the high sensitivity and negative predictive value of the clinical assessment performed by an experienced toxicologist, and the high specificity and positive predictive value that characterize urinary amanitin analysis. PMID- 15533031 TI - Paradoxical cerebral cortical hyperexcitability following flupirtine overdose. AB - We report the case of a patient with increased cerebral cortical excitability following intoxication with flupirtine, a centrally acting analgesic and antispastic drug. Typically, flupirtine exerts membrane stabilizing and hyperpolarizing effects through activation of neuronal G-protein regulated inwardly rectifying potassium channels (GIRK). Based on these properties, GIRK activators have been suggested as candidates for antiepileptic drug development. In contrast, our observation suggests that these substances may also display unexpected proconvulsant effects in vivo. PMID- 15533032 TI - Lack of toxic effects following acute overdose of cellcept (mycophenolate mofetil). AB - Mycophenolate mofetil is an immunosuppressive drug used for prevention of graft rejection following solid organ transplant and for treatment of autoimmune disorders. We report a case of a 24-year-old female with lupus nephritis that presented following ingestion of 10 grams of mycophenolate in a suicide gesture. Serum levels confirmed ingestion. The patient was treated with decontamination and supportive care and recovered with no adverse effect. PMID- 15533033 TI - Neuroleptic malignant syndrome in a child treated with an atypical antipsychotic. AB - Neuroleptic malignant syndrome (NMS) is an uncommon potentially fatal side effect of neuroleptic drugs, characterized by movement disorder, altered mental status and autonomic instability. A single dose of clotiapine was administered to an 11 year old male with acute psychosis. The previously healthy child had signs consistent with NMS including hyperthermia, hypertension, motor and mental changes. Repeat examination performed two weeks later, demonstrated that while his hyperthermia subsided, his mental status deteriorated. Olanzapine was administered, after which the child had hyperthermia, dystonia and more pronounced restlessness, once again consistent with NMS. He developed respiratory failure and was intubated and mechanically ventilated. Lorazepam, dantrolene and bromocriptine were administered as treatment of possible NMS. His mental condition, movement disorder and autonomic dysfunction improved significantly. Two weeks later, the patient was discharged in good general condition without the need for any ongoing medical treatment. There are only few case reports of NMS in children treated with olanzapine, an atypical antipsychotic. In children, caution must be exercised when prescribing antipsychotics, particularly atypical antipsychotics as these drugs may cause NMS. Because of the low incidence of NMS, a high index of suspicion is needed to identify cases so prompt treatment can be undertaken. PMID- 15533034 TI - Acute endosulfan poisoning with cerebral edema and cardiac failure. AB - BACKGROUND: Organochlorine insecticides are highly toxic compounds that are responsible for a number of severe intoxications worldwide with several deaths. Despite their widespread use in agriculture during the 1940s to 1960s and the well-known signs and symptoms of intoxication, the clinical picture in case of poisoning varies. We report two cases of acute intentional endosulfan intoxication with cerebral edema and cardiac failure. CASE REPORTS: Both cases developed life-threatening signs like epileptic state, respiratory insufficiency and hemodynamic instability soon after ingestion. The survivor developed severe myocardial insufficiency and pulmonary edema documented by echocardiography and x ray of the chest. The deceased patient developed severe cerebral edema and multiorgan failure ten days after ingestion of Thiodan 35. The peak serum concentration of endosulfan in the survivor was 0.12 mg/L approximately 23 hours after ingestion, whereas the peak blood concentration in the fatal case was 0.86 mg/L approximately 25 hours post-ingestion. Post-mortem endosulfan levels in different organs were determined. CONCLUSION: Endosulfan is a highly toxic organochlorine insecticide that produces well-known neurological symptoms of tonic-clonic convulsions, headache, dizziness and ataxia but also can cause gastrointestinal symptoms and metabolic disturbances. Life-threatening cerebral edema and hemodynamic instability may occur. Treatment is symptomatic and supportive. PMID- 15533035 TI - DNA polymerase fidelity: kinetics, structure, and checkpoints. AB - On careful examination of existing kinetic data for correct and incorrect dNTP incorporations by a variety of DNA polymerases, it is apparent that these enzymes resist a unified description. Instead, the picture that emerges is a rather complex one: for most polymerases, there is evidence for a noncovalent step preceding phosphoryl transfer, but there are less reliable data for determining whether the noncovalent step or phosphoryl transfer is rate-limiting during misincorporation. Although the structural conservation in the polymerase superfamily is probably reflected in a common set of intermediates along the reaction pathway, the energetics of these species vary even when closely related polymerases are compared. Consequently, some polymerases apparently show more discrimination between correctly paired and mispaired dNTPs in the binding step, and polymerases may differ in terms of which step of the reaction is rate limiting in correct and incorrect insertion reactions. Because of the higher energy barrier in the misincorporation reaction, at least some of the intermediates both before and after the rate-limiting step in the misincorporation pathway will have higher energies than the corresponding intermediates in correct incorporation; consequently, these steps can serve as kinetic checkpoints. PMID- 15533036 TI - Structural examination of phi-value analysis in protein folding. AB - Protein folding intermediates and transition states are commonly characterized using a protein engineering procedure (Phi-value analysis) based on several assumptions, including (1) intermediates and transition states have native-like conformations and (2) single mutations from larger hydrophobic residues to smaller ones do not perturb their structures. Although Phi-value analysis has been widely used, these assumptions have not been tested to date because of the lack of high-resolution structures of intermediates and transition states. We recently have determined the structure of a folding intermediate for a four-helix bundle protein (Rd-apocytochrome b(562)) using NMR. The intermediate has the N terminal helix unfolded. The other three helices fold in a native-like topology with extensive non-native hydrophobic interactions. Here, we have determined the Phi values for 14 hydrophobic core residues, including those with significant non native interactions. All of the Phi values are in the normal range from 0 to 1, indicating that these non-native interactions cannot be identified by the common Phi-value analysis, and therefore, the first assumption is not valid for this intermediate. We also determined the structure of a mutant (F65A) of the intermediate and found that the structure of the intermediate is not perturbed by the mutation, supporting the second assumption. Together, these results suggest that Phi-value analysis may be valid for characterizing the energetics of the interactions between the mutated residue and others, but not for determining the detailed structures of intermediates and transition states because non-native interactions may exist and may not be identifiable by the common Phi-value analysis. PMID- 15533037 TI - Assignment of endogenous substrates to enzymes by global metabolite profiling. AB - Enzymes regulate biological processes through the conversion of specific substrates to products. Therefore, of fundamental interest for every enzyme is the elucidation of its natural substrates. Here, we describe a general strategy for identifying endogenous substrates of enzymes by untargeted liquid chromatography-mass spectrometry (LC-MS) analysis of tissue metabolomes from wild type and enzyme-inactivated organisms. We use this method to discover several brain lipids regulated by the mammalian enzyme fatty acid amide hydrolase (FAAH) in vivo, including known signaling molecules (e.g., the endogenous cannabinoid anandamide) and a novel family of nervous system-enriched natural products, the taurine-conjugated fatty acids. Remarkably, the relative hydrolytic activity that FAAH exhibited for lipid metabolites in vitro was not predictive of the identity of specific FAAH substrates in vivo. Thus, global metabolite profiling establishes unanticipated connections between the proteome and metabolome that enable assignment of an enzyme's unique biochemical functions in vivo. PMID- 15533038 TI - Intracellular uptake and inhibition of gene expression by PNAs and PNA-peptide conjugates. AB - Peptide nucleic acids (PNAs) offer a distinct option for silencing gene expression in mammalian cells. However, the full value of PNAs has not been realized, and the rules governing the recognition of cellular targets by PNAs remain obscure. Here we examine the uptake of PNAs and PNA-peptide conjugates by immortal and primary human cells and compare peptide-mediated and DNA/lipid mediated delivery strategies. We find that both peptide-mediated and lipid mediated delivery strategies promote entry of PNA and PNA-peptide conjugates into cells. Confocal microscopy reveals a punctate distribution of PNA and PNA-peptide conjugates regardless of the delivery strategy used. Peptide D(AAKK)(4) and a peptide containing a nuclear localization sequence (NLS) promote the spontaneous delivery of antisense PNAs into cultured cells. The PNA-D(AAKK)(4) conjugate inhibits expression of human caveolin 1 (hCav-1) in both HeLa and primary endothelial cells. DNA/lipid-mediated delivery requires less PNA, while peptide mediated delivery is simpler and is less toxic to primary cells. The ability of PNA-peptide conjugates to enter primary and immortal human cells and inhibit gene expression supports the use of PNAs as antisense agents for investigating the roles of proteins in cells. Both DNA/lipid-mediated and peptide-mediated delivery strategies are efficient, but the compartmentalized localization of PNAs suggests that improving the cellular distribution may lead to increased efficacy. PMID- 15533039 TI - Positive selection dictates the choice between kinetic and thermodynamic protein folding and stability in subtilases. AB - Subtilisin E (SbtE) is a member of the ubiquitous superfamily of serine proteases called subtilases and serves as a model for understanding propeptide-mediated protein folding mechanisms. Unlike most proteins that adopt thermodynamically stable conformations, the native state of SbtE is trapped into a kinetically stable conformation. While kinetic stability offers distinct functional advantages to the native state, the constraints that dictate the selection between kinetic and thermodynamic folding and stability remain unknown. Using highly conserved subtilases, we demonstrate that adaptive evolution of sequence dictates selection of folding pathways. Intracellular and extracellular serine proteases (ISPs and ESPs, respectively) constitute two subfamilies within the family of subtilases that have highly conserved sequences, structures, and catalytic activities. Our studies on the folding pathways of subtilisin E (SbtE), an ESP, and its homologue intracellular serine protease 1 (ISP1), an ISP, show that although topology, contact order, and hydrophobicity that drive protein folding reactions are conserved, ISP1 and SbtE fold through significantly different pathways and kinetics. While SbtE absolutely requires the propeptide to fold into a kinetically trapped conformer, ISP1 folds to a thermodynamically stable state more than 1 million times faster and independent of a propeptide. Furthermore, kinetics establish that ISP1 and SbtE fold through different intermediate states. An evolutionary analysis of folding constraints in subtilases suggests that observed differences in folding pathways may be mediated through positive selection of specific residues that map mostly onto the protein surface. Together, our results demonstrate that closely related subtilases can fold through distinct pathways and mechanisms, and suggest that fine sequence details can dictate the choice between kinetic and thermodynamic folding and stability. PMID- 15533040 TI - Hydrophobic helical hairpins: design and packing interactions in membrane environments. AB - Helix-helix interactions within membranes are dominated by van der Waals packing motifs and side chain-side chain hydrogen bond formation, which act in tandem to determine the residues that comprise the interface between two given helices. To explore in a systematic manner the tertiary contacts between transmembrane helices, we have designed and expressed in Escherichia coli highly hydrophobic helix-loop-helix constructs of prototypic sequence K(1)KKKKKKFAIAIAIIAWAX(19)AIIAIAIAIKSPGSKIAIAIAIIAZ(44)AWAIIAIAIAFKKKKKKK(62), where "small" (Ala) and "large" (Ile) residues were used to maximize the tertiary contact area. Evidence that the two transmembrane (TM) segments in the AI construct contain an interface conducive for folding into a hairpin structure was obtained from the results that (i) the single TM AI(pep) peptide derived from the AI hairpin forms SDS-resistant dimers on PAGE gels and (ii) the corresponding sequence forms a strong dimer when examined in vivo in TOXCAT assays. Site directed mutagenesis of AI hairpins was carried out to incorporate each of the 20 commonly occurring amino acids at X positions. Analysis on Western blots using an oligomerization assay in 12% NuPage-sodium dodecyl sulfate (SDS) indicated that mutants with X = E, D, Q, R, N, H, and K largely formed SDS-resistant dimers which likely correspond to H-bonded four-helix bundles-while all the others (e.g., X = F, W, L, I, M, V, C, Y, A, T, S, G, and P) remained monomeric. Systematic studies of X/Z double mutants indicated that formation of hairpin dimers is the result of the disruption of stabilizing interactions between the antiparallel helices within the AI construct. The overall results suggest that, in situations where hydrophobic van der Waals packing energy between helices is sufficient to prevent significant rotation about the major axes of interacting helices, intrahairpin side chain-side chain H-bond formation will occur mainly when pairs of polar residues are interfacially located and proximal. Knowledge of the relative contributions of these forces should be of value, for example, in clarifying the context--and the structural consequences--of disease-related mutations. PMID- 15533041 TI - ATR-FTIR spectroscopy and isotope labeling of the PM intermediate of Paracoccus denitrificans cytochrome c oxidase. AB - The structure of the P(M) intermediate of Paracoccus denitrificans cytochrome c oxidase was investigated by perfusion-induced attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. Transitions from the oxidized to P(M) state were initiated by perfusion with CO/oxygen buffer, and the extent of conversion was quantitated by simultaneously monitoring visible absorption changes. In prior work, tentative assignments of bands were proposed for heme a(3), a change in the environment of the protonated state of a carboxylic acid, and a covalently linked histidine-tyrosine ligand to Cu(B) that has been found in the catalytic site. In this work, reduced minus oxidized difference spectra at pH 6.5 and 9.0 and P(M) minus oxidized difference spectra at pH 9.0 were compared in unlabeled, universally (15)N-labeled, and tyrosine-ring-d(4)-labeled proteins to improve these assignments. In the reduced minus oxidized difference spectrum, (15)N labeling resulted in large changes in the amide II region and a 9 cm(-1) downshift in a 1105 cm(-1) trough that is attributed to histidine. In contrast, changes induced by tyrosine-ring-d(4) labeling were barely detectable where the isotope-sensitive bands are expected. Both isotope substitutions had large effects on P(M) minus oxidized difference spectra. A prominent trough at 1542 cm( 1) was shifted to 1527 cm(-1) with (15)N labeling, and its magnitude was diminished with the appearance of a 1438 cm(-1) trough with tyrosine-ring-d(4) labeling. Both isotope substitutions also had large effects on a 1314 cm(-1) trough in the same spectra. These shifts indicate that the bands are linked to both a nitrogenous compound and a tyrosine, the most obvious candidate being the covalent histidine-tyrosine ligand of Cu(B). Comparison with model material data suggests that the tyrosine hydroxyl group is protonated when the binuclear center is oxidized but deprotonated in the P(M) intermediate. Positive bands at 1519 and 1570 cm(-1) were replaced with bands at 1504 and 1556 cm(-1), respectively, with tyrosine-ring-d(4) labeling, are characteristic of upsilon(7a)(C-O) and upsilon(C C) bands of neutral phenolic radicals, and most likely reflect the formation of the neutral radical state of the histidine-tyrosine ligand in P(M). PMID- 15533042 TI - Trapped tyrosyl radical populations in modified reaction centers from Rhodobacter sphaeroides. AB - The photosynthetic reaction center from the purple bacterium Rhodobacter sphaeroides has been modified such that the bacteriochlorophyll dimer, when it becomes oxidized after light excitation, is capable of oxidizing tyrosine residues. One factor in this ability is a high oxidation-reduction midpoint potential for the dimer, although the location and protein environment of the tyrosine residue appear to be critical as well. These factors were tested in a series of mutants, each of which contains changes, at residues L131, M160, M197, and M210, that give rise to a bacteriochlorophyll dimer with a midpoint potential of at least 800 mV. The protein environment was altered near tyrosine residues that are either present in the wild type or introduced by mutagenesis, focusing on residues that could act as acceptors for the phenolic proton of the tyrosine upon oxidation. These mutations include Ser M190 to His, which is near Tyr L162, the combination of His M193 to Tyr and Arg M164 to His, which adds a Tyr-His pair, and the combinations of Arg L135 to Tyr with Tyr L164 to His, Arg L135 to Tyr with Tyr L144 to Glu, and Arg L135 to Tyr with Tyr L164 to Phe. Radicals were produced in the mutants by using light to initiate electron transfer. The radicals were trapped by freezing the samples, and the relative populations of the oxidized dimer and tyrosyl radicals were determined by analysis of low temperature electron paramagnetic resonance spectra. The mutants all showed evidence of tyrosyl radical formation at high pH, and the extent of radical formation at Tyr L135 with pH differed depending on the identity of L144 and L164. The results show that tyrosine residues within approximately 10 A of the dimer can become oxidized when provided with a suitable protein environment. PMID- 15533043 TI - Catalytic mechanism of guanidinoacetate methyltransferase: crystal structures of guanidinoacetate methyltransferase ternary complexes. AB - Guanidinoacetate methyltransferase (GAMT) is the enzyme that catalyzes the last step of creatine biosynthesis. The enzyme is found in abundance in the livers of all vertebrates. The intact GAMT from recombinant rat liver has been crystallized with an inhibitor S-adenosylhomocysteine (SAH) and a substrate guanidinoacetate (GAA), and with SAH and an inhibitor guanidine (GUN). These ternary complex structures have been determined at 2.0 A resolution. GAMT has an alpha/beta open sandwich structure, and the N-terminal section (residues 1-42) covers the active site entrance so that the active site is not visible. SAH has extensive interactions with GAMT through H-bonds and hydrophobic interactions. The guanidino groups of GAA and GUN form two pairs of H-bonds with E45 and D134, respectively. The carboxylate group of GAA interacts with the backbone amide groups of L170 and T171. A model structure of GAMT containing the two substrates (SAM and GAA) was built by attaching a methyl group (C(E)) on S(D) of the bound SAH. On the basis of this model structure, a catalytic mechanism of GAMT is proposed. The active site entrance is opened when the N-terminal section is moved out. GAA and SAM enter the active site and interact with the amino acid residues on the surface of the active site by polar and nonpolar interactions. O(D1) of D134 and C(E) of SAM approach N(E) of GAA from the tetrahedral directions. The O(D1)...N(E) and C(E)...N(E) distances are 2.9 and 2.2 A, respectively. It is proposed that three slightly negatively charged carbonyl oxygen atoms (O of T135, O of C168, and O(B) of GAA) around O(D1) of D134 increase the pK(a) of O(D1) so that O(D1) abstracts the proton on N(E). A strong nucleophile is generated on the deprotonated N(E) of GAA, which abstracts the methyl group (C(E)) from the positively charged S(D) of SAM, and creatine (methyl-GAA) and SAH (demethyl-SAM) are produced. E45, D134, and Y221 mutagenesis studies support the proposed mechanism. A mutagenesis study and the inhibitory mechanism of guanidine analogues support the proposed mechanism. PMID- 15533044 TI - Site-specific effects of zinc on the activity of family II pyrophosphatase. AB - Family II pyrophosphatases (PPases), recently found in bacteria and archaebacteria, are Mn(2+)-containing metalloenzymes with two metal-binding subsites (M1 and M2) in the active site. These PPases can use a number of other divalent metal ions as the cofactor but are inactive with Zn(2+), which is known to be a good cofactor for family I PPases. We report here that the Mg(2+)-bound form of the family II PPase from Streptococcus gordonii is nearly instantly activated by incubation with equimolar Zn(2+), but the activity thereafter decays on a time scale of minutes. The activation of the Mn(2+)-form by Zn(2+) was slower but persisted for hours, whereas activation was not observed with the Ca(2+)- and apo-forms. The bound Zn(2+) could be removed from PPase by prolonged EDTA treatment, with a complete recovery of activity. On the basis of the effect of Zn(2+) on PPase dimerization, the Zn(2+) binding constant appeared to be as low as 10(-12) M for S. gordonii PPase. Similar effects of Zn(2+) and EDTA were observed with the Mg(2+)- and apo-forms of Streptococcus mutans and Bacillus subtilis PPases. The effects of Zn(2+) on the apo- and Mg(2+)-forms of HQ97 and DE15 B. subtilis PPase variants (modified M2 subsite) but not of HQ9 variant (modified M1 subsite) were similar to that for the Mn(2+)-form of wild-type PPase. These findings can be explained by assuming that (a) the PPase tightly binds Mg(2+) and Mn(2+) at the M2 subsite; (b) the activation of the corresponding holoenzymes by Zn(2+) results from its binding to the M1 subsite; and (c) the subsequent inactivation of Mg(2+)-PPase results from Zn(2+) migration to the M2 subsite. The inability of Zn(2+) to activate apo-PPase suggests that Zn(2+) binds more tightly to M2 than to M1, allowing direct binding to M2. Zn(2+) is thus an efficient cofactor at subsite M1 but not at subsite M2. PMID- 15533045 TI - Structural studies of metal ions in family II pyrophosphatases: the requirement for a Janus ion. AB - Family II inorganic pyrophosphatases (PPases) constitute a new evolutionary group of PPases, with a different fold and mechanism than the common family I enzyme; they are related to the "DHH" family of phosphoesterases. Biochemical studies have shown that Mn(2+) and Co(2+) preferentially activate family II PPases; Mg(2+) partially activates; and Zn(2+) can either activate or inhibit (Zyryanov et al., Biochemistry, 43, 14395-14402, accompanying paper in this issue). The three solved family II PPase structures did not explain the differences between the PPase families nor the metal ion differences described above. We therefore solved three new family II PPase structures: Bacillus subtilis PPase (Bs-PPase) dimer core bound to Mn(2+) at 1.3 A resolution, and, at 2.05 A resolution, metal free Bs-PPase and Streptococcus gordonii (Sg-PPase) containing sulfate and Zn(2+). Comparison of the new and old structures of various family II PPases demonstrates why the family II enzyme prefers Mn(2+) or Co(2+), as an activator rather than Mg(2+). Both M1 and M2 undergo significant changes upon substrate binding, changing from five-coordinate to octahedral geometry. Mn(2+) and Co(2+), which readily adopt different coordination states and geometries, are thus favored. Combining our structures with biochemical data, we identified M2 as the high-affinity metal site. Zn(2+) activates in the M1 site, where octahedral geometry is not essential for catalysis, but inhibits in the M2 site, because it is unable to assume octahedral geometry but remains trigonal bipyramidal. Finally, we propose that Lys205-Gln81-Gln80 form a hydrophilic channel to speed product release from the active site. PMID- 15533046 TI - Role of lysine-256 in Citrobacter freundii tyrosine phenol-lyase in monovalent cation activation. AB - Tyrosine phenol-lyase (TPL) from Citrobacter freundii is dependent on monovalent cations, K(+) or NH(4)(+), for high activity. We have shown previously that Glu 69, which is a ligand to the bound cation, is important in monovalent cation binding and activation [Sundararaju, B., Chen, H., Shillcutt, S., and Phillips, R. S. (2000) Biochemistry 39, 8546-8555]. Lys-256 is located in the monovalent cation binding site of TPL, where it forms a hydrogen bond with a structural water bound to the cation. This lysine residue is highly conserved in sequences of TPL and the paralogue, tryptophan indole-lyase. We have now prepared K256A, K256H, K256R, and E69D/K256R mutant TPLs to probe the role of Lys-256 in monovalent cation binding and activation. K256A and K256H TPLs have low activity (k(cat)/K(m) values of 0.01-0.1%), are not activated by monovalent cations, and do not exhibit fluorescence emission at 500 nm from the PLP cofactor. In contrast, K256R TPL has higher activity (k(cat)/K(m) about 10% of wild-type TPL), is activated by K(+), and exhibits fluorescence emission from the PLP cofactor. K256A, K256H, and K256R TPLs bind PLP somewhat weaker than wild-type TPL. E69D/K256R TPL was prepared to determine if the guanidine side chain could substitute for the monovalent cation. This mutant TPL has wild-type activity with S-Et-L-Cys or S-(o-nitrophenyl)-L-Cys but has no detectable activity with L-Tyr. E69D/K256R TPL is not activated by monovalent cations and does not show PLP fluorescence. In contrast to wild-type and other mutant TPLs, PLP binding to E69D/K256R is very slow, requiring several hours of incubation to obtain 1 mol of PLP per subunit. Thus, E69D/K256R TPL appears to have altered dynamics. All of the mutant TPLs react with inhibitors, L-Ala, L-Met, and L-Phe, to form equilibrating mixtures of external aldimine and quinonoid intermediates. Thus, Lys-256 is not the base which removes the alpha-proton during catalysis. The results show that the function of Lys-256 in TPL is in monovalent cation binding and activation. PMID- 15533047 TI - Domain architecture of the p62 subunit from the human transcription/repair factor TFIIH deduced by limited proteolysis and mass spectrometry analysis. AB - TFIIH is a multiprotein complex that plays a central role in both transcription and DNA repair. The subunit p62 is a structural component of the TFIIH core that is known to interact with VP16, p53, Eralpha, and E2F1 in the context of activated transcription, as well as with the endonuclease XPG in DNA repair. We used limited proteolysis experiments coupled to mass spectrometry to define structural domains within the conserved N-terminal part of the molecule. The first domain identified resulted from spontaneous proteolysis and corresponds to residues 1-108. The second domain encompasses residues 186-240, and biophysical characterization by fluorescence studies and NMR analysis indicated that it is at least partially folded and thus may correspond to a structural entity. This module contains a region of high sequence conservation with an invariant FWxxPhiPhi motif (Phi representing either tyrosine or phenylalanine), which was also found in other protein families and could play a key role as a protein protein recognition module within TFIIH. The approach used in this study is general and can be straightforwardly applied to other multidomain proteins and/or multiprotein assemblies. PMID- 15533048 TI - Evolution of glutamate dehydrogenase regulation of insulin homeostasis is an example of molecular exaptation. AB - Glutamate dehydrogenase (GDH) is found in all organisms and catalyzes the oxidative deamination of glutamate to 2-oxoglutarate. While this enzyme does not exhibit allosteric regulation in plants, bacteria, or fungi, its activity is tightly controlled by a number of compounds in mammals. We have previously shown that this regulation plays an important role in insulin homeostasis in humans and evolved concomitantly with a 48-residue "antenna" structure. As shown here, the antenna and some of the allosteric regulation first appears in the Ciliates. This primitive regulation is mediated by fatty acids and likely reflects the gradual movement of fatty acid oxidation from the peroxisomes to the mitochondria as the Ciliates evolved away from plants, fungi, and other protists. Mutagenesis studies where the antenna is deleted support this contention by demonstrating that the antenna is essential for fatty acid regulation. When the antenna from the Ciliates is spliced onto human GDH, it was found to fully communicate all aspects of mammalian regulation. Therefore, we propose that glutamate dehydrogenase regulation of insulin secretion is a example of exaptation at the molecular level where the antenna and associated fatty acid regulation was created to accommodate the changes in organelle function in the Ciliates and then later used to link amino acid catabolism and/or regulation of intracellular glutamate/glutamine levels in the pancreatic beta cells with insulin homeostasis in mammals. PMID- 15533049 TI - Effects of methylphosphonate, a phosphate analogue, on the expression and degradation of the high-affinity phosphate transporter Pho84, in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, the Pho84 high-affinity transport system is the major phosphate transporter activated when the cells experience a limitation in external phosphate. In this study, we have compared the phosphate-responsive mechanism of cells expressing PHO84 with a Deltapho84 strain by use of a phosphate analogue, methylphosphonate, which was judged to be suitable for assessment of phosphate homeostasis in the cells. Intracellular levels of the analogue, which in several respects mimicks phosphate, were monitored by (31)P NMR spectroscopy. Results show that methylphosphonate is a nonhydrolyzable and nonutilizable analogue that cannot be used to replenish phosphate or polyphosphate in yeast cells grown under conditions of phosphate limitation. However, the presence of methylphosphonate under such conditions represses the Pho5 acidic phosphatase activity of PHO84 cells, a finding that implies a direct role of the analogue in the regulation of phosphate-responsive genes and/or proteins. Likewise, accumulation of the Pho84 protein at the plasma membrane of the same cells is inhibited by methylphosphonate, although the derepressive expression of the PHO84 gene is unperturbed. Thus, a post-transcriptional regulation is suggested. Supportive of this suggestion is the fact that addition of methylphosphonate to cells with abundant and active Pho84 at the plasma membrane causes enhanced internalization of the Pho84 protein. Altogether, these observations suggest that the Pho84 transporter is regulated not only at the transcriptional level but also by a direct molecule-sensing mechanism at the protein level. PMID- 15533050 TI - Inhibition of islet amyloid polypeptide fibril formation: a potential role for heteroaromatic interactions. AB - The formation of amyloid fibril is associated with major human diseases, including Alzheimer's disease, prion diseases, and type 2 diabetes. Methods for efficient inhibition of amyloid fibril formation are therefore highly clinically important. A principal approach for the inhibition of amyloid formation is based on the use of modified molecular recognition elements. Here, we demonstrate efficient inhibition of amyloid formation of the type 2 diabetes-related human islet amyloid polypeptide (hIAPP) by a modified aromatic peptide fragment and a small aromatic polyphenol molecule. A molecular recognition assay using peptide array analysis suggested that molecular recognition between hIAPP and its core amyloidogenic module is mediated by aromatic rather than hydrophobic interactions. To study the possible effect of aromatic interactions on inhibition of hIAPP fibril formation, we have used peptide and small molecule inhibitors. The addition of a nonamyloidogenic peptide analogue of the core module NFGAILSS, in which phenylalanine was substituted with tyrosine (NYGAILSS), resulted in substantial inhibition of fibril formation by hIAPP. The inhibition was significantly stronger than the one achieved using a beta-sheet breaker conjugated peptide NFGAILPP. On the basis of the molecular arrangement of the tyrosine-phenylalanine interaction, we suggest that the inhibition stems from the geometrical constrains of the heteroaromatic benzene-phenol interaction. In line with this notion, we demonstrate remarkable inhibition of hIAPP fibril formation and cytotoxicity toward pancreatic beta-cells by a small polyphenol molecule, the nontoxic phenol red compound. Taken together, our results provide further experimental support for the potential role of aromatic interactions in amyloid formation and establish a novel approach for its inhibition. PMID- 15533051 TI - A series of related nucleotide analogues that aids optimization of fluorescence signals in probing the mechanism of P-loop ATPases, such as actomyosin. AB - We have synthesized a set of ATP and ADP analogues that have a fluorophore linked to the nucleotide via the 3'-position of the ribose moiety. Combinations of three different coumarins are each attached via different length linkers. A linker based on propylenediamine increases the separation between the nucleotide and fluorophore relative to that of the previously reported ethylenediamine-linked coumarin nucleotides [Webb, M. R., and Corrie, J. E. T. (2001) Biophys. J. 81, 1562-1569]. A synthesis of 3'-amino-3'-deoxyATP is described using a combination of chemical and enzymatic procedures, mostly from published methods for synthesis of this compound but with some modifications that improved the convenience of the experimental procedures. This compound is used as a basis of a series of analogues with effectively a zero-length linker. Fluorescence properties of all these analogues are described, together with the kinetics of their interaction with rabbit skeletal myosin subfragment 1 in the presence and absence of actin. One particular analogue, deac-aminoATP [3'-(7-diethylaminocoumarin-3 carbonylamino)-3'-deoxyadenosine 5'-triphosphate], shows a 17-fold enhancement of fluorescence upon binding to this (skeletal) myosin II. As the diphosphate, it exhibits a large signal change upon dissociation from the actomyosin, with kinetics similar to those of natural ADP. The ability of this set of analogues to produce large signals indicated potential uses when scarce proteins are studied in small amounts. PMID- 15533052 TI - Uncovering the basis for nonideal behavior of biological molecules. AB - The molecular origin of the nonideal behavior for concentrated binary solutions of biochemical compounds is examined. The difference between activities expressed in the molar and molal conventions can be large. Considering the range from dilute to concentrated, we show that molar activity coefficients can be represented by simple but rigorous equations involving between one and three parameters only. We derive a universal relationship interconverting the scales of molarity and molality without requiring the density of the solution. The equations are developed from first principles using a statistical thermodynamic theory of molar activity coefficients. It is shown how to express activity coefficients in different concentration scales, and the advantages and disadvantages of using certain scales are discussed and compared with the experimental data. Several classes of biochemically relevant compounds, many of which are naturally occurring osmolytes, are discussed: six saccharides (glucose, xylose, maltose, mannose, raffinose, and sucrose), four polyols (glycerol, mannitol, erythritol, and sorbitol), five amino acids (glycine, alanine, sarcosine, glycine betaine, and proline), and urea. Of the 16 solutes, 10 could be described in terms of a single parameter that is due to pure first-order effects (packing, hydration, or space limitation). The remaining six exhibit significant second-order effects (solute-solute interactions) and require two additional parameters, one typically identified with the volume occupied per solute molecule in the pure solute (crystal or liquid) and the other with a self association constant. The activity coefficients of the osmolytes roughly display the rank order found with respect to their ability to stabilize proteins. These findings are discussed in terms of the physical principles that give rise to the activity coefficients. PMID- 15533054 TI - Inhibition of wild-type and mutant human immunodeficiency virus type 1 proteases by GW0385 and other arylsulfonamides. AB - The arylsulfonamide derivatives described herein were such potent inhibitors of human immunodeficiency virus type 1 (HIV-1) protease (enzyme, E) that values for the inhibition constants (K(i)) could not be determined by conventional steady state kinetic techniques (i.e., the minimal enzyme concentration usable for the activity assay was much greater than the value of the dissociation constant). Consequently, two alternative methods were developed for estimation of K(i) values. The first method employed kinetic determinations of values for k(1) and k(-1), from which K(i) was determined (k(-1)/k(1)). The second method was a competitive displacement assay used to determine binding affinities of other inhibitors relative to that of GW0385. In these assays, the inhibitor of unknown affinity was used to displace [(3)H]GW0385 from E.[(3)H]GW0385. From the concentration of E.[(3)H]GW0385 at equilibrium, the concentrations of enzyme bound and free inhibitors were calculated, and the ratio of the K(i) value of the unknown to that of GW0385 was determined (K(i,unknown)/K(i,GW0385)). The values of k(1) were calculated from data in which changes in the intrinsic protein fluorescence of the enzyme associated with inhibitor binding were directly or indirectly monitored. In the case of saquinavir, the fluorescence changes associated with complex formation were large enough to monitor directly. The value of k(1) for saquinavir was 62 +/- 2 microM(-1) s(-1). In the case of GW0385, the fluorescence changes associated with complex formation were too small to monitor directly. Consequently, the value of k(1) was estimated from a competition experiment in which the effect of GW0385 on the binding of E to saquinavir was determined. The value of k(1) for GW0385 was estimated from these experiments to be 137 +/- 4 microM(-1) s(-1). Because E.[(3)H]GW0385 was stable in the standard buffer at room temperature for greater than 33 days, the value of the first-order rate constant for dissociation of E.[(3)H]GW0385 (k(-1)) could be estimated from the time-course for exchange of E.[(3)H]GW0385 with excess unlabeled GW0385. The value of k(-1) calculated from these data was (2.1 +/- 0.1) x10(-6) s(-1) (t(1/2) = 91 h). The K(i) value of wild-type HIV-1 protease for GW0385, calculated from these values for k(1) and k(-1), was 15 +/- 1 fM. Three multidrug resistant enzymes had K(i) values for GW0385 that were less than 5 pM. PMID- 15533053 TI - Dioxygenases without requirement for cofactors and their chemical model reaction: compulsory order ternary complex mechanism of 1H-3-hydroxy-4-oxoquinaldine 2,4 dioxygenase involving general base catalysis by histidine 251 and single-electron oxidation of the substrate dianion. AB - 1H-3-Hydroxy-4-oxoquinaldine 2,4-dioxygenase (Hod) is a cofactor-less dioxygenase belonging to the alpha/beta hydrolase fold family, catalyzing the cleavage of 1H 3-hydroxy-4-oxoquinaldine (I) and 1H-3-hydroxy-4-oxoquinoline (II) to N-acetyl- and N-formylanthranilate, respectively, and carbon monoxide. Bisubstrate steady state kinetics and product inhibition patterns of HodC, the C69A protein variant of Hod, suggested a compulsory-order ternary-complex mechanism, in which binding of the organic substrate precedes dioxygen binding, and carbon monoxide is released first. The specificity constants, k(cat)/K(m,A) and k(cat)/K(m,O)()2, were 1.4 x 10(8) and 3.0 x 10(5) M(-1) s(-1) with I and 1.2 x 10(5) and 0.41 x 10(5) M(-1) s(-1) with II, respectively. Whereas HodC catalyzes formation of the dianion of its organic substrate prior to dioxygen binding, HodC-H251A does not, suggesting that H251, which aligns with the histidine of the catalytic triad of the alpha/beta hydrolases, acts as general base in catalysis. Investigation of base-catalyzed dioxygenolysis of I by electron paramagnetic resonance (EPR) spectroscopy revealed formation of a resonance-stabilized radical upon exposure to dioxygen. Since in D(2)O spectral properties are not affected, exchangeable protons are not involved, confirming that the dianion is the reactive intermediate that undergoes single-electron oxidation. We suggest that in the ternary complex of the enzyme, direct single-electron transfer from the substrate dianion to dioxygen may occur, resulting in a radical pair. Based on the estimated spin distribution within the radical anion (observed in the model reaction of I), radical recombination may produce a C4- or C2 hydroperoxy(di)anion. Subsequent intramolecular attack would result in the 2,4 endoperoxy (di)anion that may collapse to the reaction products. PMID- 15533055 TI - Transversal and lateral exciton energy transfer in grana thylakoids of spinach. AB - The excitation energy transfer between photosystem (PS) II complexes was studied in isolated grana disks and thylakoids using chlorophyll a fluorescence induction measurements in the presence of DCMU under stacked and destacked conditions. Destacking of grana was achieved using a sonication protocol in a buffer without MgCl(2). The degree of stacking was controlled and quantified by atomic force microscopy and by the concomitant absorption changes. As expected from the literature, intact thylakoids showed a strong dependency of the connectivity of PSII centers, the F(m)/F(o) ratio as well as the fraction of PSIIbeta centers on the MgCl(2) concentration. In contrast, these parameters did not change in isolated grana disks. In particular, the connectivity remained constantly high irrespective of the degree of destacking. These differences were explained by the high protein density in grana disks, which hinders separation and mixing of proteins sufficiently to change energy transfer properties. Due to the occurrence of stroma lamella in intact thylakoids, intermixing of PSII and PSI is possible and allows for changes in F(m)/F(o) ratio as is the separation of LHCII from PSII, thus leading to an increase in the fraction of PSIIbeta. Even if mixing and separation of proteins are impaired in isolated grana disks, destacking should lead to a decrease in connectivity if transversal excitation energy transfer between two opposite membranes is significant. Because the connectivity is constant over all degrees of destacking employed, we conclude that the energy transfer in granas is mainly lateral. PMID- 15533056 TI - Oxalate-inducible AMBP gene and its regulatory mechanism in renal tubular epithelial cells. AB - The AMBP [A1M (alpha1-microglobulin)/bikunin precursor] gene encodes two plasma glycoproteins: A1M, an immunosuppressive lipocalin, and bikunin, a member of plasma serine proteinase inhibitor family with prototypical Kunitz-type domain. Although previously believed to be constitutively expressed exclusively in liver, the present study demonstrates the induction of this gene by oxalate in porcine proximal tubular LLC-PK1 cells and rat kidney. In liver, the precursor protein is cleaved in the Golgi network by a furin-like enzyme to release constituent proteins, which undergo glycosylation before their export from the cell. In the renal tubular cells, A1M and bikunin co-precipitate, indicating lack of cleavage of the precursor protein. As the expression of the AMBP gene is regulated by A1M specific cis elements and transcription factors, A1M protein was studied as a representative of AMBP gene expression in renal cells. Oxalate treatment (500 microM) resulted in a time- and dose-dependent induction of A1M protein in LLC PK1 cells. Of the four transcription factors, HNF-4 (hepatocyte nuclear factor-4) has been reported previously to be a major regulator of AMBP gene expression in liver. Electrophoretic mobility-shift assay, supershift assay, immunoreactivity assay and transfection-based studies showed the presence of an HNF-4 or an HNF-4 like protein in the kidney, which can affect the expression of the AMBP gene. In situ hybridization and immunocytochemical studies showed that the expression of this gene in kidney was mainly restricted to cells lining the renal tubular system. PMID- 15533057 TI - Expression and regulation of sterol 27-hydroxylase (CYP27A1) in human macrophages: a role for RXR and PPARgamma ligands. AB - CYP27A1 (sterol 27-hydroxylase) catalyses an important sterol elimination pathway in the human macrophage, and consequently may protect against atherosclerosis. We studied the expression and regulation of CYP27A1 in a human macrophage-like cell line, THP-1, and primary HMDMs (human monocyte-derived macrophages). In both macrophage cell types, we found that CYP27A1 expression is independent of cellular cholesterol levels and of LXR (liver X receptor)-dependent control of transcription. However, the RXR (retinoid X receptor) ligand, 9-cis-retinoic acid, upregulates CYP27A1 expression. Of the RXR heterodimeric partners tested, PPAR (peroxisome-proliferator-activated receptor) gamma ligands significantly increased CYP27A1 mRNA levels. Its reversal by a PPARgamma antagonist demonstrated the specificity of this effect. Interestingly, HMDMs express markedly higher levels of CYP27A1 than THP-1 macrophages, and this difference was reflected in both protein levels and enzyme activities between the two cell types. In conclusion, stimulation of CYP27A1 by PPARgamma may represent a key previously unrecognized mechanism by which PPARgamma protects against atherosclerosis. PMID- 15533058 TI - Juvenile hyaline fibromatosis: a case report and review of the literature. AB - BACKGROUND: Juvenile hyaline fibromatosis (JHF) is a rare, inherited condition characterized by tumor-like growth of hyalinized fibrous tissue on the head and neck, joint contractures, and gingival hypertrophy. There may be marked clinical heterogeneity. METHODS: We present a case of a 3-year-old Haitian boy with multiple firm nodules on the scalp and chin without joint contractures or gingival hypertrophy. Family history was not available. RESULTS: Biopsy specimens from three scalp nodules were processed with routine and immunohistochemical stains. The matrix was periodic acid Schiff (PAS) and Alcian blue positive. The cellular stromal component was positive for vimentin and scattered factor XIIIa positive cells were found. Osteoclast-like giant cells were also noted, and stained for CD68. CONCLUSIONS: Our patient had the nodular growths on the scalp and face that are characteristically found in JHF. Microscopic examination confirmed the diagnosis and showed scattered intracytoplasmic and extracellular eosinophilic globules in three separate biopsy specimens. These were positive with PAS. PMID- 15533059 TI - Pyoderma gangrenosum: clinicopathologic correlation and proposed diagnostic criteria. AB - Pyoderma gangrenosum is a rare but significant cause of ulcerations. It is a diagnosis of exclusion. Herein, we suggest diagnostic criteria and some historical perspectives on the diagnosis of pyoderma gangrenosum. PMID- 15533060 TI - Asiaticoside induction for cell-cycle progression, proliferation and collagen synthesis in human dermal fibroblasts. AB - Asiaticoside, isolated from Centella asiatica, promotes fibroblast proliferation and extracullar matrix synthesis in wound healing. The precise mechanism, however, in molecular and gene expression levels still remains partially understood. Using cDNA microarray technology, the alternation of genes expression profiles was determined in a human dermal fibroblast in vitro in the presence of asiaticoside (30 microg/ml). Fifty-four genes, with known functions for cell proliferation, cell-cycle progression and synthesis of the extracellular matrix, were significantly up-regulated in our "whole-genes nest" expression profile at various timepoints. Furthermore, mRNA levels and protein productions of certain genes responsible for extracellular matrix (ECM) synthesis (e.g. encoding type I and type III collagen proteins) were evaluated by Northern blot and radioimmunoassay, respectively. As a result, there is a close correlation among the gene profile, mRNA and protein production in the cells response to asiaticoside stimulation. This information could be used for exploring the target genes in response to asiaticoside in fibroblasts. PMID- 15533061 TI - Importance of the direct fresh examination in the diagnosis of cutaneous sporotrichosis. AB - BACKGROUND: To date the direct fresh examination has not been considered, except by a few authors, as a valid laboratory procedure for the diagnosis of cutaneous sporotrichosis. METHODS: Forty-two patients with cutaneous sporotrichosis (36 lymphangitic and six fixed) were studied. Pus was obtained through digital compression of opened lesions followed by collection with a spatula and examination of the material between the slide and the coverslip. Cultures on Sabouraud's glucose agar slants with cloramphenicol and cicloheximide were made in all cases. RESULTS: Direct, fresh examination asteroid bodies of Sporothrix schenckii were found in 85.7% of the cases, which allowed the authors to immediately diagnose the disease. Cultures confirmed the diagnosis in 95.2% of the patients. CONCLUSIONS: Fastness, simplicity and low cost of the fresh examination lead us to recommend systematic use of the method for primary diagnosis of the disease, mainly at first level public health laboratories. PMID- 15533062 TI - What patients with vitiligo believe about their condition. AB - BACKGROUND: Vitiligo is a common disease with an unknown etiology. It is a chronic disease which causes severe disfigurement, and so may significantly affect a patient's quality of life. This study was designed to evaluate the illness perception and quality of life in patients with vitiligo. PATIENTS AND METHODS: A questionnaire consisting of 25 questions on the causes, timeline, consequences, and control of disease was given to 80 patients with vitiligo attending a private skin clinic in Tehran, Iran, in 1999. The roles of age, sex, duration of disease, education, extent of disease, and family history of vitiligo were assessed. RESULTS: Eighty patients (22 males and 57 females, one not mentioned), with a mean age of 30 years (SD = 11 years) and a disease duration of 7.1 years (SD = 7.8 years), completed the questionnaire; 62.5% of the patients believed that stress played a role in their disease; 31.3% believed that their genetic background played a role in their disease, more so in patients with a positive family history of vitiligo. Almost one-half of patients believed that their illness had major consequences on their lives. One-half of patients believed that their illness was likely to be permanent rather than temporary, more so in patients with a longer duration of disease and a higher level of education. Only 60% of patients considered that their treatments were effective. CONCLUSION: Vitiligo may considerably affect various aspects of patients' lives. The knowledge of patients about the causes and course of their disease is limited. PMID- 15533063 TI - Lupus erythematosus tumidus. AB - BACKGROUND: Lupus erythematosus tumidus (LET) is a rare form of chronic cutaneous lupus erythematosus that characteristically presents as a succulent, erythematous plaques on sun-exposed areas. The histopathological change that primarily distinguishes LET from other variants of cutaneous lupus erythematosus is the lack of alterations of the dermo-epidermal junction and epidermis. Our purpose was to describe 15 cases of LET from Thailand. METHODS: The records of 15 patients diagnosed with LET at Srinagarind Hospital Medical School between 1995 and 2002 were reviewed on the following aspects: clinical manifestation, laboratory investigation, direct immunofluorescence study, phototesting, histopathological findings, clinical course, and treatment. RESULTS: All 15 patients consisted of 10 females and five males. The mean age at onset was 38.8 years. The lesions more commonly affect the face and trunk. One patient had concomitant discoid lupus erythematosus and two developed systemic involvement 4 and 6 months after LET. Direct immunofluorescence studies all showed negative results. Positive phototest reactions were detected in five of 10 tested patients: four with ultraviolet A and B and one with ultraviolet A only. Most patients responded well to chloroquine phosphate and the remainder successfully improved with methotrexate, topical and systemic corticosteroids. CONCLUSION: Our data demonstrate the features of LET that are repeatable and characteristic. Lupus erythematosus tumidus in this study was more common in women, and photosensitivity reactions of the patients were seen less frequently compared with those in the earlier report of Kuhn et al. Furthermore, two of 15 patients exhibited systemic involvement in the course of their disease. PMID- 15533064 TI - A quantitative study of epidermal Langerhans cells in cutaneous leishmaniasis caused by Leishmania tropica. AB - OBJECTIVE: The purpose of this study was to characterize the number and distribution of epidermal Langerhans cells in different clinical forms of dry type cutaneous leishmaniasis (CL). METHODS: Sixteen cases of dry-type cutaneous leishmaniasis caused by Leishmania tropica were studied. These cases were classified clinically as five cases of acute leishmaniasis with indurated papules, nodules and plaques with central crust formation and duration < 2 years, six cases of lupoid leishmaniasis with characteristic papules around previous scars of cutaneous leishmaniasis with duration > 2 years, and five cases of chronic nonlupoid type with nonhealing lesions of duration > 2 years. Paraffin embedded blocks were stained with hematoxylin and eosin (H&E) and stained immunohistochemically for CD1a. RESULTS: The number of Langerhans cells per millimeter length of epidermis was increased in acute cases compared to chronic and lupoid cases. CONCLUSIONS: Lesions of acute leishmaniasis contain the greatest amounts of antigen for presentation, so Langerhans cells increase in number and in trafficking to present antigens derived from Leishman bodies to the cellular immune system. In chronic leishmaniasis, the Langerhans cell population is reduced, perhaps because of exhaustion of the source of Langerhans cells, or because of reduced response to modified antigen. PMID- 15533065 TI - Sporotrichoid nocardiosis caused by Nocardia nova in a patient infected with human immunodeficiency virus. PMID- 15533066 TI - Atypical clinical form of cutaneous leishmaniasis: erysipeloid form. PMID- 15533067 TI - Management of diaper dermatitis. PMID- 15533068 TI - Extensive Darier's disease with esophageal involvement. AB - Although the involvement of mucous membranes in Darier's disease is relatively uncommon, Darier's disease has been associated with mucosal lesions, especially in the oral mucosa. In the English literature there is only one report describing the involvement of esophageal mucous membranes in a patient with Darier's disease. Herein, we report the second case of Darier's disease with esophageal involvement. PMID- 15533069 TI - Lupus erythematosus-like lesions in a carrier of X-linked chronic granulomatous disease: a case report and personal considerations. PMID- 15533070 TI - Chronic telogen effluvium or early androgenetic alopecia? PMID- 15533071 TI - Round excision of small, benign, papular and dome-shaped melanocytic nevi on the face. AB - BACKGROUND: Patients frequently request the removal of benign, papular and dome shaped nevi for cosmetic or functional reasons. Total excision is probably the most widely used method of removal. AIM: To introduce the round excision technique for the treatment of benign, papular and dome-shaped nevi on the face. METHODS: In a prospective study, 36 benign, papular or dome-shaped nevi of all types were removed by the round excision technique. The lesions were circumscribed with a number 15 scalpel blade, 2 mm beyond the limits, with incision to the full depth of the dermis, and removed by cutting horizontally at the maximum depth of the circular incision. Histologic examinations were performed for all specimens. RESULTS: Complete removal of nevi was achieved in all patients with excellent or good cosmetic results. Of the 36 nevi, 24 were intradermal and 12 were compound nevi. Dog-ear formation was observed in only one patient. CONCLUSION: Round excision may be a better alternative to conventional fusiform or shave excision of benign, dome-shaped or papular nevi of the face. PMID- 15533072 TI - Prospective, noncomparative open study from Kuwait of the role of intravenous immunoglobulin in the treatment of toxic epidermal necrolysis. AB - BACKGROUND: High-dose intravenous immunoglobulin (IVIG) is emerging as a promising new therapy for treating the rare but potentially fatal drug reaction toxic epidermal necrolysis (TEN). Experimental in vitro studies support that IVIG can block the Fas-FasL-mediated apoptosis in TEN. METHODS: Twelve consecutive patients (7M, 5F) with TEN admitted over a 5-year period from January 1998 to December 2002 were treated with a dose of 0.5-1.0 g/kg/d of IVIG for 4-5 days along with standard care protocol. Clinical outcome in terms of average duration to arrest the progression, complete healing, hospital stay, side-effects and complications were determined to find the efficacy of IVIG treatment. RESULTS: Average age was 27.16 years (7-50 years). There were four children (2M, 2F) aged 7-12 years. One patient had an underlying malignancy. No patient had HIV infection. The average total body surface area involvement was 57.5% (30-90%). An IVIG infusion was started, on average, 1.58 days (1-3 days) after admission. All patients responded well to the treatment. There was no mortality. The disease progression was arrested in a mean of 2.83 days (1-5 days). Time taken for complete healing (re-epithelialization) was 7.33 days (5-13 days). The average duration of hospital stay was 12.5 days (7-21 days). No side-effects of the IVIG treatment were observed in these patients. The drugs triggering TEN in these patients were phenytoin (four patients), followed by penicillin (three), cotrimoxazole (two), phenobarbital and furosemide (one patient each), respectively. In one patient, the offending drug could not be ascertained. CONCLUSION: Our experience of treating 12 patients with TEN using IVIG, in Kuwait, confirms that it is a safe and effective treatment for these patients. PMID- 15533073 TI - Generalized necrobiosis lipoidica treated with a combination of split-thickness autografting and immunomodulatory therapy. PMID- 15533074 TI - Mouse-related dermatosis. PMID- 15533075 TI - Round excision of small, benign, papular and dome-shaped melanocytic nevi on the face. PMID- 15533076 TI - Topical calcipotriol in childhood vitiligo: an Indian experience. PMID- 15533077 TI - Discoid lupus erythematosus recurs after plastic surgery. PMID- 15533078 TI - Pyoderma Gangrenosum: clinicopathological correlation and proposed diagnostic criteria. PMID- 15533079 TI - Minority groups: an outdated concept? PMID- 15533081 TI - Changing professional practice in tuberculosis care: an educational intervention. AB - AIM: This paper describes the development and implementation of an experiential, participatory in-service education programme focusing on patient-centeredness, problem-solving and critical reflection for primary providers delivering care to tuberculosis patients in South Africa. BACKGROUND: Tuberculosis is a major contributor to the disease burden in developing countries. In South Africa, approximately 90% of tuberculosis patients are managed by nurses at the primary level. Despite efforts to improve quality of care for these patients, many fail to complete their treatment as prescribed. Poor rapport between health care providers and patients with tuberculosis is a major reason for non-adherence to treatment. Methods of improving the quality of care delivery and communication between health care providers and patients with tuberculosis is therefore a priority. DISCUSSION: The paper outlines the stages of developing and implementing the education programme and reflects on this process. Data is drawn from an in-depth qualitative evaluation of the delivery and impacts of the intervention. The approach was acceptable to health care providers and adaptable to the needs of specific clinics. Participants evaluated the educational intervention positively, noting that it facilitated critical reflection on their work; encouraged problem-solving; and heightened their awareness of communication with patients and with colleagues. However, important structural barriers to practice change were identified, including conditions of service, relations with colleagues and support from management. CONCLUSIONS: Experiential, participatory in-service education can be implemented on a large scale in primary care settings. However, the process is resource intensive and the impacts of such education may be limited by barriers at other levels of the health system. PMID- 15533082 TI - Altered body image: appearance-related concerns of people with visible disfigurement. AB - AIM: The aim of this paper is to report a study to establish the extent and type of psychosocial needs of outpatients attending for treatment of a wide range of disfiguring conditions. BACKGROUND: Visible disfigurements can be associated with extensive psychosocial difficulties. The majority of research to date has been carried out with people identified by themselves or others as experiencing difficulties. Little is known about levels of distress in the broader population of patients receiving treatment for a range of disfiguring conditions. METHODS: A cross-sectional survey was conducted, with a convenience sample. Participants (n = 458) drawn from 15 outpatient clinics completed standardized measures of anxiety and depression (Hospital Anxiety and Depression Scale), social anxiety and avoidance (Derriford Appearance Scale short-form) and quality of life (World Health Organization Quality of Life Brief Scale). A semi-structured interview was used to generate further quantitative and qualitative data about individual concerns, and satisfaction with the provision of care. Staff views about levels of psychosocial distress were elicited through group discussions. RESULTS: The results revealed high levels of psychological distress in the sample, compared with normative values. The majority of difficulties related to problems experienced in social situations. Patient satisfaction with care was generally high; however, and 71% of participants expressed a moderate to strong desire for a health care professional with training to deal with their appearance-related concerns. Nursing staff felt unable to address patients' appearance-related difficulties because of time constraints, lack of an environment conducive to the discussion of patients' concerns, and lack of appropriate knowledge and training. CONCLUSION: A significant proportion of participants experienced psychosocial distress in relation to their visible difference. Psychosocial needs were poorly met in current outpatient care provision, and a range of options could be considered to address these more effectively. PMID- 15533083 TI - Factors affecting quality of oral care in intensive care units. AB - AIMS: This paper presents a study to assess to nurses' attitudes and practices concerning oral care and to determine predictors of the quality of oral care in intensive care units. BACKGROUND: The oropharynx of critically ill patients becomes colonized with potential respiratory pathogens; oral care has been shown to reduce oropharyngeal bacteria and ventilator-associated pneumonia. METHODS: In April 2002, a random and national sample of 420 intensive care unit directors was asked to participate in the survey. Of invited directors, 126 (30%) agreed to participate and were sent questionnaires to be completed anonymously by their staff, and 102 institutions returned 556 surveys. This gave a response rate of 83% of those who consented to participate. RESULTS: The path model shows that nurses' oral care education, having sufficient time to provide care, prioritizing oral care, and not viewing oral care as unpleasant had direct effects on the quality of provided care. Intensive care unit experience, oral care education, and having sufficient time had indirect effects. CONCLUSION: Improving the quality of oral care in intensive care units is a multi-layered task. Reinforcing proper oral care in education programmes, de-sensitizing nurses to the often perceived unpleasantness of cleaning oral cavities, and working with hospital managers to allow sufficient time to attend to oral care are recommended. PMID- 15533084 TI - Predictors of health behaviours in college students. AB - AIM: This paper reports a study examining the direct effects of perceived stress, perceived availability of and satisfaction with social support, and self efficacy, and examines the intermediary roles of perceived threat (perceived susceptibility x perceived severity), benefits, and barriers on alcohol behaviour, smoking behaviour, physical activity and nutrition behaviour, general safety behaviour and sun-protective behaviour in college students. BACKGROUND: Health behaviours formed during young adulthood may have a sustaining impact on health across later life. Entering college can be an exciting, yet stressful event for many adolescents and young adults as they face trying to adapt to changes in academic workloads, support networks, and their new environment. Coupled with these changes and new-found responsibilities, they have greater freedom and control over their lifestyles than ever before. However, researchers have shown globally that many college students engage in various risky health behaviours. METHOD: A cross-sectional sample of 161 college students enrolled in an introductory psychology course completed self-report questionnaires regarding stress; social support; self-efficacy; and components of the Health Belief Model including perceived threat, perceived benefits, perceived barriers; and common health behaviours. Step-wise multiple regression analysis was conducted and significant predictors were retained as modifiers in the path analysis. FINDINGS: Self-efficacy significantly predicted alcohol and smoking behaviour, physical activity and nutrition protective behaviour, general safety protective behaviour and sun-protective behaviour. Under high-perceived threat, self-efficacy was mediated by perceived barriers for binge drinking and moderated by perceived barriers for physical activity and nutrition behaviours. In addition, under high perceived threat, self-efficacy was moderated by perceived threat for alcohol use at 30 days and 6 months. Under low threat, self-efficacy was mediated by perceived barriers for smoking behaviour and general safety protective behaviours. CONCLUSIONS: Future health promotion programmes with college students must use interventions that maximize self-efficacy and ultimately reduce barriers to adopting a healthy lifestyle. PMID- 15533085 TI - Nurse-patient relationships in palliative care. AB - AIM: The aim of this paper is to report a study exploring aspects of nurse patient relationships in the context of palliative care. BACKGROUND: Although there are numerous studies addressing nurse-patient relationships, little research has focused on these in the context of palliative are. Furthermore, no previous study has examined the relationship in the Chinese context. METHODS: Qualitative data were collected from 10 hospice nurses and 10 terminally ill patients by means of open ended unstructured interviews. Respondents were asked to reflect on practices and incidents that would allow an understanding of the meaning of nurse-patient relationships in palliative care. RESULTS: Four major categories emerged from the perspectives of patients and nurses: (1) forming a relationship of trust; (2) being part of the family; (3) refilling with fuel along the journey of living and dying; and (4) enriched experiences. Responses revealed that a relationship of trust is formed, and that nurses are not only regarded as health professionals, but also become part of the family or a good friend. Nurses who develop trusting relationships demonstrate a holistic approach to caring, show their understanding of patients' suffering, are aware of their unvoiced needs, provide comfort without actually being asked, and are reliable, proficient, competent and dedicated in their care. CONCLUSION: Trust, the achievement of the goals of patients and nurses, caring and reciprocity are important elements of nurse-patient relationships in palliative care. Such relationships not only improve patients' physical and emotional state, but also facilitate their adjustment to their illness, ease pain and can ultimately lead to a good death experience. It is nurses' personal qualities and skills, which are embedded in these relationships, that constitute excellence in nursing care. Nurses also derive satisfaction and are enriched through the relationships. PMID- 15533086 TI - Chronic illness self-management: locating the 'self'. AB - AIM: In this paper, we present the findings of a recent research project in which we explored self- management with older people who were diagnosed with asthma. BACKGROUND: Asthma self-management literature has focused on the need for the patient to 'adhere' to prescribed therapies, in particular the taking of medications, monitoring of respiratory function or recognizing and avoiding triggers. METHOD: Data were generated during a period of 9 months from three sources; in-depth interviews with 24 older participants, an open-ended questionnaire and two mixed-gender participatory action research groups. FINDINGS: Based on current literature, our previous research findings which have 'unpacked' what is 'self'-management, and data generated in this project, we propose that three asthma management models are in operation: Medical Model of Self-management, Collaborative Model of Self-management and Self-Agency Model of Self-management. Locating the 'self' in self-management means acknowledging that many people living with a chronic condition are already self-determining and their expertise should be acknowledged as such. CONCLUSION: Health care professionals can best facilitate people toward self-agency by embracing new understandings of self-management in long-term illness. This process is enhanced when the expertise a person brings to the management of their condition is given the respect it deserves. There needs to be a focus on providing people with the means to grow and learn in a participative relationship that cannot be fully realized with 'off the shelf' self-management solutions. PMID- 15533087 TI - Relationship between two anxiety instruments in patients receiving mechanical ventilatory support. AB - AIMS: The primary aim of this paper is to discuss the relationship between the Visual Analog Scale-Anxiety (VAS-A) and the Spielberger State Anxiety Inventory (SAI) in patients receiving mechanical ventilatory support. A secondary aim is to provide suggestions for the nurse-researcher to consider when selecting an instrument to measure anxiety. BACKGROUND: Anxiety is a common experience for critically ill patients receiving mechanical ventilatory support. It is a challenge, however, for nurse-researcher to select an instrument to measure anxiety that is valid and reliable yet does not cause great response burden for participants. Visual analog scales may reduce response burden, but lack sound validation in research participants receiving mechanical ventilatory support. METHODS: This study used a correlational design. A convenience sample of critically ill patients receiving mechanical ventilatory support (n = 200) were asked to rate their current level of anxiety on the 20-item Spielberger SAI and a 100-mm VAS-A. RESULTS: Eight participants were unable to complete the Spielberger SAI; 100% completed the VAS-A. The two instruments were found to be significantly correlated at r = 0.50; P = 0.01. CONCLUSION: The VAS-A was found to be less burdensome for research participants than the Spielberger SAI, resulting in no missing data on the VAS-A. Findings from this study provide initial validation of the VAS-A as a justifiable instrument to measure anxiety in patients receiving mechanical ventilatory support. Researchers are advised to balance reliability and validity properties with response burden when selecting an instrument to measure anxiety in patients with communication challenges and energy limitations. PMID- 15533088 TI - Problem-solving competency of nursing graduates. AB - AIM: This paper reports a study describing and evaluating the outcomes of problem based learning (PBL) programmes in nursing schools in South Africa in terms of the competence of graduates to solve problem in actual clinical settings, and comparing this competence with that of graduates from non-PBL programmes. BACKGROUND: The nursing literature tends to equate problem-solving with patient centred problems or the nursing process. However, it is also a skill used in managing the work role, working in a team and managing a health care unit. Problem-solving refers to the process of selectively attending to information in a patient care setting. The investigation of problem-solving in nursing is complicated by the complex relationships between different cognitive processes. METHODS: A qualitative evaluation study, descriptive and comparative in nature, was carried out. In-depth interviews were held with graduates and their supervisors, asking them to identify problem-solving incidents in which they had been involved. Template analysis style and Benner's interpretive approach were used to analyse the data. FINDINGS: The majority of the incidents described by the graduates (84%) were graded at the advanced beginner level or above. The majority of incidents at the novice level came from the non-PBL group. 'Using people skills' and 'being assertive' were the two problem-solving strategies most often used. The PBL group fared better than the non-problem-based group in the level of their problem-solving ability. CONCLUSION: The findings of this study suggest that further research is warranted into the problem-solving abilities of PBL graduates, their personal development over time and at different stages of practice. In addition, it would be interesting to follow the development of their problem-solving abilities over time. PMID- 15533089 TI - Developing concept maps from problem-based learning scenario discussions. AB - AIMS: This paper reports a study examining the effects of adopting concept mapping in problem-based learning scenario discussions on the improvement of students' learning outcomes in a nursing course. BACKGROUND: Students in Taiwan usually have a high degree of anxiety about whether or not they have learned enough. Problem-based learning is a method of teaching that uses a patient situation or scenario to stimulate students to acquire and apply information to solve problems. Concept mapping can promote problem-solving and critical thinking to help students organize complex patient data, process complex relationships and offer holistic care to patients. METHODS: An experimental design was used, with participants randomly assigned either to a control or experimental group. The experimental group participated in six problem-based learning scenario discussions during the 16-week semester, while the control group attended a traditional course. RESULTS: The experimental group had significantly higher proposition and hierarchy scores for their concept maps compared with the control group. There were no significant differences in the cross-link and example scores between the two groups. In general, the total score difference between the groups did not reach statistical significance levels. Only one student in the experimental group obtained a high score; most participants in both groups (over 50%) obtained low scores. CONCLUSIONS: Concept mapping strategies may be useful for analysis of individual student's thinking processes for (1) emphasizing key concepts or main ideas, (2) understanding relationships between different concepts, including cause-effect and part-whole relationships, (3) reviewing propositions, hierarchies and cross-links in a logically scientific way, and (4) revising concept structures to agree with theory and experience. PMID- 15533090 TI - Advanced practice nursing roles: development, implementation and evaluation. AB - AIM: The aim of this paper is to discuss six issues influencing the introduction of advanced practice nursing (APN) roles: confusion about APN terminology, failure to define clearly the roles and goals, role emphasis on physician replacement/support, underutilization of all APN role domains, failure to address environmental factors that undermine the roles, and limited use of evidence-based approaches to guide their development, implementation and evaluation. BACKGROUND: Health care restructuring in many countries has led to substantial increases in the different types and number of APN roles. The extent to which these roles truly reflect advanced nursing practice is often unclear. The misuse of APN terminology, inconsistent titling and educational preparation, and misguided interpretations regarding the purpose of these roles pose barriers to realizing their full potential and impact on health. Role conflict, role overload, and variable stakeholder acceptance are frequently reported problems associated with the introduction of APN roles. DISCUSSION: Challenges associated with the introduction of APN roles suggests that greater attention to and consistent use of the terms of the terms advanced nursing practice, advancement and advanced practice nursing is required. Advanced nursing practice refers to the work or what nurses do in the role and is important for defining the specific nature and goals for introducing new APN roles. The concept of advancement further defines the multi-dimensional scope and mandate of advanced nursing practice and distinguishes differences from other types of nursing roles. Advanced practice nursing refers to the whole field, involving a variety of such roles and the environments in which they exist. Many barriers to realizing the full potential of these roles could be avoided through better planning and efforts to address environmental factors, structures, and resources that are necessary for advanced nursing practice to take place. CONCLUSIONS: Recommendations for the future introduction of APN roles can be drawn from this paper. These include the need for a collaborative, systematic and evidence-based process designed to provide data to support the need and goals for a clearly defined APN role, support a nursing orientation to advanced practice, promote full utilization of all the role domains, create environments that support role development, and provide ongoing evaluation of these roles related to predetermined goals. PMID- 15533091 TI - A framework for the introduction and evaluation of advanced practice nursing roles. AB - AIM: This paper describes a participatory, evidence-based, patient-focused process for advanced practice nursing (APN) role development, implementation, and evaluation (PEPPA framework). BACKGROUND: Despite the growing demand for advanced practice nurses, there are limited data to guide the successful implementation and optimal utilization of these roles. The participatory, evidence-based, patient-focused process, for guiding the development, implementation, and evaluation of advanced practice nursing (PEPPA) framework is an adaptation of two existing frameworks and is designed to overcome role implementation barriers through knowledge and understanding of APN roles and environments. The principles of participatory action research directed the construction of the new framework. CONCLUSIONS: The process for implementing and evaluating APN roles is as complex and dynamic as the roles themselves. The PEPPA framework is shaped by the underlying principles and values consistent with APN, namely, a focus on addressing patient health needs through the delivery of coordinated care and collaborative relationships among health care providers and systems. Engaging environmental stakeholders as participants in the process provides opportunity to identify the need and shared goals for a clearly defined APN role. The process promotes increased understanding of APN roles and optimal use of the broad range of APN knowledge, skills, and expertise in all role domains and scope of practice. The steps for planning and implementation are designed to create environments to support APN role development and long-term integration within health care systems. The goal-directed and outcome-based process also provides the basis for prospective ongoing evaluation and improvement of both the role and delivery of health care services. PMID- 15533096 TI - Intermultidisciplinarity... a new concept for nursing research. PMID- 15533097 TI - An exploration of the factors that influence the implementation of evidence into practice. AB - BACKGROUND: The challenges of implementing evidence-based practice are complex and varied. Against this background a framework has been developed to represent the multiple factors that may influence the implementation of evidence into practice. It is proposed that successful implementation is dependent upon the nature of the evidence being used, the quality of context, and, the type of facilitation required to enable the change process. This study sets out to scrutinize the elements of the framework through empirical enquiry. AIMS AND OBJECTIVES: The aim of the study was to address the following questions: * What factors do practitioners identify as the most important in enabling implementation of evidence into practice? * What are the factors practitioners identify that mediate the implementation of evidence into practice? * Do the concepts of evidence, context and facilitation constitute the key elements of a framework for getting evidence into practice? DESIGN AND METHODS: The study was conducted in two phases. Phase 1: Exploratory focus groups (n = 2) were conducted to inform the development of an interview guide. This was used with individual key informants in case study sites. Phase 2: Two sites with on-going or recent implementation projects were studied. Within sites semi-structured interviews were conducted (n = 17). RESULTS: A number of key issues in relation to the implementation of evidence into practice emerged including: the nature and role of evidence, relevance and fit with organizational and practice issues, multi professional relationships and collaboration, role of the project lead and resources. CONCLUSIONS: The results are discussed with reference to the wider literature and in relation to the on-going development of the framework. Crucially the growing body of evidence reveals that a focus on individual approaches to implementing evidence-based practice, such as skilling-up practitioners to appraise research evidence, will be ineffective by themselves. RELEVANCE TO CLINICAL PRACTICE: Key elements that require attention in implementing evidence into practice are presented and may provide a useful checklist for future implementation and evaluation projects. PMID- 15533098 TI - Reflective practice and daily ward reality: a covert power game. AB - BACKGROUND: The concept of reflection is propounded in the literature as an epistemology for practice that enables practitioners to solve their daily problematic situations through conscious thought processes which eventually leads to practice-based knowledge. Hence, reflection became a central tenet of both theory development and educational provision in nursing. Furthermore, this centrality of reflection was reinforced by statutory nursing bodies and service providers by adopting it as the means for carrying out adequate professional practice. Although this may be the case, issues of implementation of reflection within the daily reality of practitioners are frequently overlooked within the literature. Moreover, little consideration appears to be given on the impact that the organizational culture and the politics of power may exert on the implementation of reflective practices within daily ward reality. AIMS: This paper explores how reflection is viewed by nurses within their daily reality in the medical wards, examines the relationships between the organizational culture of these wards and the practitioners and investigates whether reflective methods of practice were being implemented when the study was conducted. METHODS: An interpretative ethnographical methodology was implemented and the data collecting methods used were observation, interviews and qualitative content analysis with a group of 16 practising nurses from four medical wards of one NHS Trust in England. Two interviews were conducted with each nurse within 3-week intervals. The content of these interviews evolved from the analysis of episodes of practice observed when the nurses were giving nursing care. These data were supplemented by narratives from the nurses' in the form of written reflective accounts that were analysed via qualitative content analysis techniques. FINDINGS: Four themes were generated: (i) relationships between nurses and doctors; (ii) relationships between nurses and managers; (iii) nursing practice; and (iv) nurses' input in the outcome of a clinical situation. CONCLUSIONS: The concept of reflection appears to be invalidated by the organizational hierarchy of the wards on the basis of a power struggle game. The ward structure portrays reflection as an abnormal method of practice and knowledge development. This belittlement of reflection does not mirror the practitioners' reality. Instead, it is an intelligent and intentional act on behalf of the dominant professional groups in the wards to create an illusionary picture of ward reality to allow them to survey and define nursing practice and thus maintain and remain in power. This is explicated by using Foucault's analysis and critical social theory framework. Hence, reflective processes are constrained by this covert power game; reflection, where used, is confined to nurses' personal time and space. RELEVANCE TO CLINICAL PRACTICE: The realization of this covert power game by individual clinical nurses can become the incipient point for formally using reflective methods in the practice setting. PMID- 15533099 TI - Meaning of spirituality: implications for nursing practice. AB - BACKGROUND: This research outlines some preliminary findings emerging from a grounded theory investigation into the 'meaning of spirituality'. These initial results raise some important questions about the terminology and language that nurses use regarding the term spirituality. It seems that many of the policy directives and statutory guidelines make two major assumptions regarding 'spirituality'. Firstly, patients and nurses are aware and understand the concept, and secondly, patients may require their spiritual needs to be met. These preliminary findings suggest that a dichotomy is emerging between professional assumption and patient expectation regarding the meaning of spirituality. AIM: The study had one broad research aim, to gain a deeper insight into how patients, nurses, and people from the major world religions understand the concept of 'spirituality'. DESIGN: A qualitative research design was used involving a grounded theory method of inquiry. It was felt that this qualitative method would aid the investigation of this subjective dimension of peoples' existence, enabling existing theoretical constructs and arguments to be tested. METHODS: The constant comparative method was used throughout the data collection and analysis. Analysis was undertaken at two levels, 'overview analysis' and 'line-by-line analysis'. This enabled the creation of categories and central themes. RESULTS: Constant comparative analysis resulted in the formation of several categories and central themes. Two categories presented and discussed in detail are 'definitions of spirituality', and 'diverse perceptions of spirituality'. CONCLUSION: It would seem that there is now an urgent need for nursing to evaluate and perhaps adjust its vision regarding what constitutes spirituality. Such an approach may serve to reduce the gap between policy and public expectation. RELEVANCE TO CLINICAL PRACTICE: It seems that there may be no 'precise' terminology associated with the language used to define spirituality, raising possible implications for nursing practice and nurse education. PMID- 15533100 TI - Nurses' attitudes, behaviours and perceived barriers towards pressure ulcer prevention. AB - BACKGROUND: Pressure ulcers are not a plague of modern man; they have been known to exist since ancient Egyptian times. However, despite the increasing expenditure on pressure ulcer prevention, pressure ulcers remain a major health care problem. Although nurses do not have the sole responsibility for pressure ulcer prevention, nurses have a unique opportunity to have a significant impact on this problem. AIMS AND OBJECTIVES: The specific aims of the study were to identify: * Staff nurses' attitudes towards pressure ulcer prevention. * The behaviour of staff nurses' in relation to pressure ulcer prevention. * Staff nurses' perceived barriers towards pressure ulcer prevention. DESIGN: A cross sectional survey method was used. METHODS: A randomly selected sample of staff nurses (n = 300) working in an acute care setting in an urban location was invited to participate. Data were collected using a prepiloted questionnaire. Data analysis was carried out using SPSS version 10 and SPSS Text Smart version 1.1. RESULTS: The nurses surveyed demonstrated a positive attitude towards pressure ulcer prevention. However, prevention practices were demonstrated to be haphazard and erratic and were negatively affected by lack of time and staff. These barriers prevented the nurses' positive attitude from being reflected into effective clinical practice. Education, although poorly accessed, or made available, was rarely cited as impeding practice in this area. CONCLUSION: This study suggests that positive attitudes are not enough to ensure that practice change takes place, reinforcing the complex nature of behavioural change. Implementation strategies should introduce ways in which key staff can be empowered to overcome barriers to change. RELEVANCE TO CLINICAL PRACTICE: This study provides a unique exploration of Irish nurses' attitudes, behaviours and perceived barriers towards pressure ulcer prevention, thereby contributing to the body of knowledge on this subject. As tissue viability is a new and emerging speciality, this information will contribute to evidence based practice in this area of patient care and will form the basis for the development of an educational strategy for pressure ulcer prevention and management. PMID- 15533101 TI - Inter-rater reliability of the EPUAP pressure ulcer classification system using photographs. AB - BACKGROUND: Many classification systems for grading pressure ulcers are discussed in the literature. Correct identification and classification of a pressure ulcer is important for accurate reporting of the magnitude of the problem, and for timely prevention. The reliability of pressure ulcer classification systems has rarely been tested. AIMS AND OBJECTIVES: The purpose of this paper is to examine the inter-rater reliability of classifying pressure ulcers according to the European Pressure Ulcer Advisory Panel classification system when using pressure ulcer photographs. DESIGN: Survey was among pressure ulcer experts. METHODS: Fifty-six photographs were presented to 44 pressure ulcer experts. The experts classified the lesions as normal skin, blanchable erythema, pressure ulcer (four grades) or incontinence lesion. Inter-rater reliability was calculated. RESULTS: The multirater-Kappa for the entire group of experts was 0.80 (P < 0.001). Various groups of experts obtained comparable results. Differences in classifications are mainly limited to 1 degree of difference. Incontinence lesions are most often confused with grade 2 (blisters) and grade 3 pressure ulcers (superficial pressure ulcers). CONCLUSIONS: The inter-rater reliability of the European Pressure Ulcer Advisory Panel classification appears to be good for the assessment of photographs by experts. The difference between an incontinence lesion and a blister or a superficial pressure ulcer does not always seem clear. RELEVANCE TO CLINICAL PRACTICE: The ability to determine correctly whether a lesion is a pressure ulcer lesion is important to assess the effectiveness of preventive measures. In addition, the ability to make a correct distinction between pressure ulcers and incontinence lesions is important as they require different preventive measures. A faulty classification leads to mistaken measures and negative results. Photographs can be used as a practice instrument to learn to discern pressure ulcers from incontinence lesions and to get to know the different grades of pressure ulcers. The Pressure Ulcer Classification software package has been developed to facilitate learning. PMID- 15533102 TI - Information needs of Hong Kong Chinese patients undergoing surgery. AB - BACKGROUND: The provision of information to patients is an important aspect of contemporary health care. Limitations in health resources necessitates that the provision of information is carefully planned and culturally specific to maximize the benefits to patients from the resources available. AIM AND OBJECTIVES: The purposes of the study were to recognize Chinese surgical patients' information needs on admission and ascertain why the information is important to assist in understanding how it is used and, therefore, its potential impact. METHODS: A descriptive study design was used. A convenience sample of 83 surgical patients took part comprising 51 men and 32 women. An eight-item questionnaire based on the right of patients to information as listed in the Patients' Charter in Hong Kong using a 5-point Likert scale and one open-ended question to comment on why the information was important to them was completed by patients on the day of admission. RESULTS: Patients rated highly the need for all types of information. They rated most highly the need for information about the signs and symptoms indicating postoperative complications and when to seek medical help. Patients did not rate as highly, information regarding why the doctor believes the surgery is important, treatment alternatives and explanation of the procedure. CONCLUSIONS: These findings indicate that Chinese patients are desirous of a range of relevant information. RELEVANCE TO PRACTICE: Nursing staff, in particular, need to consider the 'timeliness' of information and the cultural appropriateness of how information is delivered. PMID- 15533103 TI - Liver transplant patients: their experience in the intensive care unit. A phenomenological study. AB - BACKGROUND: The nurses' knowledge of patients' experiences contributes to gaining a deeper understanding of their health process, which will help to provide a better foundation for nursing care. The patients' experiences in the intensive care unit are singular; liver transplant patients constitute a specific group, because generally their admission to the intensive care unit marks the first step towards recovery of their health after a process in which they have lived in fear because their illness was reaching a more or less terminal stage. AIM: To describe the liver transplant patient's experience in the intensive care unit. DESIGN: Phenomenological descriptive qualitative study. METHOD: In-depth interviews were carried out with a sample of 10 patients. The interviews were audiotaped and transcribed verbatim. The transcripts were analyzed using the method devised by Giorgi and modified by Baker. RESULTS: After the data were analyzed, a general description emerged, which includes five themes reflecting the essence of what this group of patients experienced: preconceived ideas marked the way they approached transplant; captured impressions of the intensive care unit's environment and experienced sensations; their perception of the caring behaviour of the nursing staff; support from the social environment (family) and religious beliefs; and their preconceived idea of the intensive care unit contrasted with their lived experience. CONCLUSION: This research enabled us to gain in-depth knowledge of the liver transplant patient's lived experience of the intensive care unit. Nurses can optimize the nursing care plan for this group of patients based on these results. RELEVANCE TO CLINICAL PRACTICE: Awareness of the importance that these participants gave to discomfort caused in their basic needs should lead nurses to pay special attention to the nursing interventions to help with these needs. As a result of this research, nurses and other professionals in the intensive care unit know that patients have an immediate sense of positive change. This enables us to reinforce this perception, reminding them frequently that they have already had their transplant and they are progressing well, thereby aiding their recovery. With regard to the social support of liver transplant patients, nurses must facilitate the presence of family members at the patient's bedside, as participants stated that the only support they needed was that of their family. PMID- 15533104 TI - A randomized controlled trial to measure the effectiveness of a sacral wedge in preventing postoperative back pain following trans-urethral resection of the prostate (TURP) in lithotomy position. AB - BACKGROUND: Postoperative back pain is a common, yet under reported, complication of surgery. Previous studies, although small in number, have indicated that the use of a sacral wedge is effective in reducing the incidence of postoperative back pain. AIM: The aim of the study was to test the hypothesis that the intra operative use of a sacral wedge would decrease the incidence of postoperative back pain in patients undergoing trans-urethral resection of the prostate in lithotomy position. METHODS: The design of the study was a randomized controlled trial involving the use of a sacral wedge intra-operatively in a male population undergoing trans-urethral resection of the prostate. A total of 236 participants was recruited to the study and allocated to the control or intervention group by block randomization. All patients selected one of three different sized sacral wedges for use during surgery. Data were collected preoperatively, intra operatively and at postoperative days 2 and 4 and the tools included a structured questionnaire, Oswestry Disability Questionnaire, a visual analogue scale to measure pain intensity and a body map to record its location. RESULTS: Fifty-two per cent of participants reported having a history of back pain and point prevalence on admission to hospital was 27%. Twenty-eight per cent of participants experienced back pain on day 2 postoperatively and this decreased to 14% on day 4. There was no significant difference between the control and intervention groups. The severity of back pain increased after surgery. Postoperatively there were reports of back pain from participants who had not reported a history of back pain before their admission for surgery. CONCLUSIONS: In contrast to other studies we found no evidence to support the use of a sacral wedge intra-operatively to reduce the incidence of postoperative back pain. RELEVANCE TO CLINICAL PRACTICE: Despite the non-significant results, the high level of postoperative back pain stills draws attention to the need to develop strategies to reduce its incidence. PMID- 15533105 TI - Problems experienced during the first year of an acute traumatic hand injury - a prospective study. AB - BACKGROUND: Evidence-based nursing of patients with acute traumatic hand injuries treated at the hand-surgical clinic calls for knowledge about long-term implications. AIMS AND OBJECTIVES: The aim of the study was to investigate consequences of an acute traumatic hand injury during the first year after the accident. Specifically, the objectives were to investigate changes in the experience of physical and psychological problems over time, frequencies of remaining problems and the impact of the injury on work situation and life situation 1 year after the accident. DESIGN: The study was prospective and followed the patients from the first weeks to 1 year after the accident. METHOD: Ninety-one patients were assessed three times during the year. Each time the patients answered study-specific questions, the Impact of Event Scale and the Hospital Anxiety and Depression Scale. RESULTS: Problems experienced decreased during the first 3 months but tended to remain unchanged during the rest of the year. In the 1-year follow-up, the majority of the patients experienced slight or moderate functional limitations in the hand, one-third had symptoms of trauma related distress and one out of seven had troublesome pain. Half of the patients who had returned to work reported a worse work situation and 16% were still on the sick list. One-third of all patients considered their whole life situation to be worse as a result of the injury. Patients with amputations more often experienced a worse life situation. Blue-collar workers reported functional limitations and a worse life situation more often than white-collar workers. CONCLUSIONS: Patients with acute traumatic hand injuries requiring surgical treatment may experience problems, such as functional limitations, trauma-related distress and troublesome pain, with long-term implications for their work situation and life situation. RELEVANCE TO CLINICAL PRACTICE: Caring for hand injured patients should include, not only surgical treatment and other actions for restoring the function of the hand, but also preventive action and follow-up of trauma-related distress and pain. PMID- 15533106 TI - A systematic review of the effectiveness of oxygen in reducing acute myocardial ischaemia. AB - BACKGROUND: Patients with acute cardiac chest pain receive oxygen as part of their treatment. Oxygen is given in the belief it will help to correct the oxygen demand - supply imbalance of acute myocardial ischaemia. AIMS AND OBJECTIVES: The aim was to review the evidence base for the use of oxygen to treat acute myocardial ischaemia and the objective was to include all evidence of sufficient quality. DESIGN: A systematic review was carried out to provide a thorough, transparent and replicable review process. METHODS: The review included randomized and non-randomized clinical trials; patients with acute coronary syndrome (unstable angina or acute myocardial infarction); and any outcome measurements of myocardial ischaemia. RESULTS: Nine trials were found, of which two were randomized controlled trials and seven non-randomized clinical trials. There were quality assessment concerns over the methodology, size and analysis within the trials. The effectiveness of oxygen in reducing myocardial ischaemia was unclear from the review, as the trials contained data that suggested oxygen reduced myocardial ischaemia, but also data that suggested it increased myocardial ischaemia. CONCLUSIONS: No definite conclusions could be drawn as to whether oxygen reduced, increased or had no effect on acute myocardial ischaemia. The papers were divided as to recommending oxygen use for all patients with acute myocardial ischaemia or not - although all agreed that patients with systemic hypoxaemia should have this corrected by oxygen administration. The key finding of the review was that there was insufficient evidence. RELEVANCE TO CLINICAL PRACTICE: As oxygen is routinely administered to treat acute myocardial ischaemia, the lack of a clear supporting evidence base must be a source of concern. This is especially so as some of the evidence suggested oxygen may increase myocardial ischaemia. There is a need for experimental-design clinical research to test the effectiveness of oxygen in reducing myocardial ischaemia. PMID- 15533109 TI - Uncertainty in nursing practices. PMID- 15533107 TI - ICD: a qualitative study of patient experience the first year after implantation. AB - BACKGROUND: The experiences of how patients live with an implantable cardioverter defibrillator are still poorly understood. Only a few qualitative studies have investigated this phenomenon. This paper was undertaken as part of a larger project to evaluate quality of life and psychological well-being in those survivors of cardiac arrest who have received an implantable cardioverter defibrillator. AIMS AND OBJECTIVES: The aim of this qualitative study was to explore how implantable cardioverter defibrillator recipients perceive their lives during the first year after implantation of the device. METHODS: A sample of 21 patients who received an implantable cardioverter defibrillator was interviewed during three consecutive periods: one, six and 12 months postdischarge from the hospital. The semi-structured interviews were based on insights gained from a literature review. The transcripts were subjected to content analysis. RESULTS: Analysis of the data revealed seven major categories: physical deterioration, cognitive changes, perceived social support, dependency, contact with the doctor, confrontation with mortality and uncertainty surrounding having a shock. Anxiety, uncertainty, disappointment, frustration, unexpected barriers, acceptance of and dependency on the implantable cardioverter defibrillator played a major role in the lives of implantable cardioverter defibrillator recipients. Well-being improved throughout the year. During the first months after discharge from the hospital the focus was on regaining physical health. During the early postimplantation period both the implantable cardioverter defibrillator recipient and family members had adapted to the situation. Reflection on the impact and consequences of the cardiac arrest was reported more often in the late postimplantation period. CONCLUSIONS: Content analysis is a resourceful approach giving answers to questions that have hardly been addressed within the domain of cardiology. Implantable cardioverter defibrillator recipients face a complex first year, especially the first 6 months. Cognitive deterioration and confrontation with mortality are problems that need to be researched further. RELEVANCE TO CLINICAL PRACTICE: Caregivers are able to explain to future patients what they can expect in the first year after implantation. Caregivers may become more receptive to physical, psychological and social limitations and to emotional and social problems that occur in implantable cardioverter defibrillator recipients, enabling them to act upon them. PMID- 15533108 TI - Commentary on Bennett G. & Mansell I. (2004) Universal precautions: a survey of community nurses' experience and practice. Journal of Clinical Nursing 13, 413 421. PMID- 15533111 TI - Effects of training on Na, K-ATPase contents in skeletal muscle and K homeostasis of African draught bulls and cows. AB - In semiarid parts of Africa animal traction is still one of the most reliable sources for rural work power. However, draught animals have to produce most of their work power at an unfavourable moment of the year that is at the end of the dry season when feedlot is scare. To improve their condition prior to the planting season, a short training could help. The effect of training can be expressed by the changes in contents of Na(+), K(+)-pumps in the muscle cell membrane. After a training period of 15 days all cattle showed a mean increase in Na(+), K(+)-ATPase of 24% (P < 0.01) in the semitendinosus muscle of the hind leg, whereas the control group showed no change. Bulls demonstrated already after 8 days of training an increase of 20% (P < 0.05). The principal factor responsible for this up-regulation of the Na(+), K(+)-pumps is most probably the excitation of muscles during exercise. In the course of the 15 days training period, the surge of plasma K(+) in during exercise showed a tendency to decrease, but this was not significant. Nevertheless, the reduced elevations of plasma [K(+)] may delay the moment of fatigue and so improve endurance. In conclusion, a training period of 8-15 days improves the contents of Na(+), K(+) pumps and so the possible work output of draught cattle. PMID- 15533112 TI - Periventricular and subcortical leukoencephalopathy in two dachshund puppies. AB - Two wirehaired dachshund puppies were presented with generalized tremor and gait abnormalities characterized by mild ataxia, tetraparesis and slightly abnormal proprioception. Neurological examination led to the suspicion of a diffuse generalized white matter lesion. Computerized tomography and pathological examination revealed a remarkable unilateral dilatation of the lateral ventricles in each dog. Histopathological examination showed a severe reduction of stainable myelin, widespread mild perineuronal oedema with vacuolations and multifocal reactive astrocytosis affecting the subcortical and deep periventricular white, and to a lesser degree, grey matter of the cerebral hemispheres, most prominently at the level of the optic chiasm. Axons showed a moderately reduced packing density; some axons were irregularly shaped and slightly thickened. There was no evidence of myelin breakdown products and neurones appeared to be well preserved. Brain stem, cerebellum and spinal cord were normal, as was the peripheral nervous system. This leukoencephalopathy in two dachshund puppies most closely resembles human periventricular leukomalacia caused by pre- or perinatal hypoxia-ischaemia. PMID- 15533113 TI - Familial occurrence of bovine dilated cardiomyopathy in Denmark. AB - Bovine dilated cardiomyopathy (BDCM) is a hereditary disease genetically related to the Canadian Holstein sire Montwick Red Apple Sovereign (MRAS). The occurrence of this disorder in the Red Danish Dairy breed, Holsteins, and Red Holsteins in Denmark is reported. Fourteen cases were diagnosed during a 13-year period. All suffered from congestive heart failure because of progressive myocardial fibrosis. Pedigree information was available in 12 cases revealing both maternal and paternal relationship to MRAS. Several sires were identified as carriers of BDCM. These sires originated from breeding lines used to upgrade Danish cattle populations. The findings indicate that BDCM is a potential health problem for Danish cattle. PMID- 15533114 TI - Oxidative stress parameters in bitches with mammary gland tumours. AB - The aim of the present study was to describe some of the oxidative stress parameters in bitches suffering from spontaneously occurring mammary gland tumours. The experiment involved 28 bitches which had mammary gland tumours removed surgically (15 bitches with malignant tumour and 13 with benign tumour) as well as 10 clinically healthy bitches. The activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were determined in haemolysates of erythrocytes derived from the animals. The concentrations of thiobarbituric acid reactive substances (TBARS), as well as -SH groups, were determined in blood plasma. GSH-Px activity was significantly higher in the malignant tumour group than in healthy animals. SOD activity was significantly higher in animals with tumours compared with the control group. Activities of both enzymes were higher in animals with malignant tumours than in benign groups, but the differences were not statistically significant. The concentrations of TBARS and -SH groups were similar in all examined groups. The increase of antioxidative enzyme activities in these animals may suggest the activation of antioxidative defence mechanisms in mammary gland carcinogenesis. Moreover, it might indicate the participation of oxidative stress in malignancies. Further experiments involving more animals, with more frequent sample collection and the use of other oxidative stress markers are necessary. PMID- 15533115 TI - Fibrocartilaginous embolic myelopathy in a horse. AB - Fibrocartilaginous embolic myelopathy was diagnosed in a 11-year-old, male, Tennessee Walking Horse by histopathological examination of the spinal cord after the horse was killed because of severe neurological dysfunction. Both ventral funiculi of C6 and C7 cervical spinal cord had extensive necrosis with blood vessels containing fibrocartilaginous emboli. A similar fibrocartilaginous embolus was observed in a single large spinal artery adjoining the vertebral leptomeninges. PMID- 15533116 TI - Two cases of spontaneous pseudohermaphroditism in Cynomolgus monkeys (Macaca fascicularis). AB - Disorders of genital development occur in all mammals. Hermaphroditism is a condition where the subject has genital organs of both sexes. True hermaphrodites have both ovarian and testicular tissue. Pseudohermaphrodites have only one type of gonadal tissue according to which they are classified as male or female pseudohermaphrodites. Two cases of spontaneously occurring pseudohermaphroditism in Cynomolgus monkeys (Macaca fascicularis) were seen here during 1999-2001. Both animals had female external genitalia, but each was found to have testicular tissue. PMID- 15533117 TI - Lymphosarcoma in a boar. AB - A 14-month-old Yorkshire boar was examined. In the abdomen, two big tumour masses were found in the intestines. In addition, pale nodules occurred in the liver, kidneys, in and upon the spleen, in the wall of the heart and in the testicles. By immunohistochemistry tumour cells were positive for CD3, but negative for B lymphocyte antigen and CD79. From these findings it was concluded that the boar was suffering from multicentric lymphosarcoma probably of T-cell origin. PMID- 15533118 TI - Obstructive, granulomatous tracheitis caused by Onchocerca sp. in a dog. AB - A 3-year-old spayed female dog was admitted with a history of episodes of dyspnoea and coughing. Severe tracheal stenosis was found on radiography. The dog died during an episode of refractory dyspnoea. Necropsy revealed an obstruction of the thoracic part of the trachea because of a chronic granulomatous inflammation protruding into the tracheal lumen. Histological examination revealed nematodes, which were identified as Onchocerca sp. according to their morphological characteristics. In contrast to the common ocular manifestation in dogs, obstructive tracheitis caused by Onchocerca infection has not been reported before. PMID- 15533119 TI - Primary gastric histiocytic sarcoma in a dog--a case report. AB - A 12-year-old intact female mixed breed dog was presented for chronic, intermittent vomiting and diarrhoea. On endoscopic examination a protruding mass arising from the mucosal surface of the pyloric region was detected. Cytological and histological examination revealed an accumulation of pleomorphic round/oval phagocytic cells suggesting histiocytic origin. This was confirmed by immunohistochemistry. No extra-gastric involvement was detected on clinical examination or at necropsy. This is the first report of primary gastric histiocytic sarcoma in a dog. PMID- 15533120 TI - Actinomycosis in a one-humped camel (Camelus dromedarius). AB - An actinomycotic granuloma caused by Actinomyces viscosus is reported in a dromedary camel. Two hard, cutaneous, large granulamatous nodules were present on both sides of the postero-ventral side of the mandible exhibiting exudation and necrosis. After radical excision of the lesion, the daily treatment with penicillin-streptomycin combination was continued for 4 weeks. About 8 and 24 weeks from the initial treatment, no new nodules were noticed. PMID- 15533121 TI - Technical and economic evaluation of different methods of Newcastle disease vaccine administration. AB - Two types of locally produced live vaccines (HB1 and La Sota--lentogenic strains) and inactivated oil adjuvant (IOAV) vaccine were used to compare the efficiency of three vaccination techniques, namely drinking water, ocular and spray on broiler chicks. The ocular route of vaccination on 1-day-old chicks followed by a booster dose on the third week through the same route induced a significantly higher level of haemagglutination inhibition antibody titre (P < 0.0001). The highest mean antibody titre was log(2) 6.6 and 93.3% of the chicks were protected from the challenge. The spray technique induced a lower antibody titre (peak of log(2) 5.9) and only 53% of the chicks in this treatment survived against the challenge. The results of this study show that the ocular route is superior to the drinking water route, which is superior to the spray technique. The economic analysis result showed that the ocular HB1 and La Sota vaccine administration method to 1- and 21-day-old chicks gave the highest revenue followed by the drinking water method. In terms of total cost, the injection method required the highest cost (0.21 birr/chick) followed by the ocular method (0.18 birr/chick). The marginal cost of vaccine administration is too small compared with marginal revenues from relative effectiveness of the methods. The internal rate of return for the ocular method was very high. The results of sensitive analysis on revenues from different vaccination methods indicate that a 25% reduction in broiler price reduces the marginal revenue from the ocular method by 12 487 birr but this still does not prove that the ocular method is economically viable for small- and medium-scale poultry farms. PMID- 15533122 TI - An analgesic evaluation of isoxsuprine in horses. AB - Isoxsuprine is used clinically to treat navicular disease and laminitis in horses. Although it is thought to increase digital and laminar blood flow, isoxsuprine's mechanism of action remains controversial, and analgesia has been suggested recently as such possible mechanism. This research investigated the analgesic potential of isoxsuprine in healthy horses submitted to a mechanical nociceptive test. Isoxsuprine (1.2 mg/kg), xylazine (1.1 mg/kg), distilled water : ethanol 95% (2 : 1, v/v, 20 ml) and saline (0.9%, 20 ml) were injected intravenously, and nociceptive thresholds were measured over 90 min. Only xylazine significantly increased nociceptive thresholds, confirming that alpha(2) adrenoceptor agonists produce analgesia in horses. Our results do not support an analgesic mechanism of action for isoxsuprine in horses, suggesting that other mechanisms might account for the clinical efficacy of this drug or that mechanical nociceptive testing may not be sufficiently sensitive to demonstrate an analgesic effect for this drug. PMID- 15533123 TI - Development of canine systemic lupus erythematosus model. AB - The purpose of this study was to develop a model for canine systemic lupus erythematosus. Systemic lupus erythematosus (SLE) is a systemic autoimmune syndrome defined by clinical and serological features, including arthritis, glomerulonephritis, dermatitis and autoantibodies. SLE was induced in eight normal dogs by immunization with heparan sulphate, the major glycosaminoglycan of the glomerular basement membrane. All the heparan sulphate-immunized dogs showed mild-to-moderate levels of proteinuria and skin disease. Cutaneous signs associated with SLE including alopecia, erythema, crusting, scaling and seborrhoea were observed. Immunohistological examination of the skin lesions revealed deposition of immunoglobulin M and complement in the dermal-epidermal junction. Three of eight dogs showed lameness. The antinuclear antibody tests were positive with the antibody titres higher than 1:128. Therefore, this experimental SLE model could be useful for studying immune-mediated skin disease and autoimmunity. PMID- 15533124 TI - A systematic review of the survival and complication rates of fixed partial dentures (FPDs) after an observation period of at least 5 years. AB - OBJECTIVES: The objective of this systematic review was to assess the 5- and 10 year survival of implant supported fixed partial dentures (FPDs) and to describe the incidence of biological and technical complications. METHODS: An electronic MEDLINE search complemented by manual searching was conducted to identify prospective and retrospective cohort studies on FPDs with a mean follow-up time of at least 5 years. Patients had to have been examined clinically at the follow up visit. Assessment of the identified studies and data abstraction was performed independently by two reviewers. Failure and complication rates were analyzed using random-effects Poisson regression models to obtain summary estimates of 5- and 10-year survival proportions. RESULTS: The search provided 3844 titles and 560 abstracts. Full-text analysis was performed for 176 articles resulting in 21 studies that met the inclusion criteria. Meta-analysis of these studies indicated an estimated survival of implants in implant-supported FPDs of 95.4% (95 percent confidence interval (95% CI): 93.9-96.5%) after 5 and 92.8% (95% CI: 90-94.8%) after 10 years. The survival rate of FPDs supported by implants was 95% (95% CI: 92.2-96.8%) after 5 and 86.7% (95% CI: 82.8-89.8%) after 10 years of function. Only 61.3% (95% CI: 55.3-66.8%) of the patients were free of any complications after 5 years. Peri-implantitis and soft tissue complications occurred in 8.6% (95% CI: 5.1-14.1%) of FPDs after 5 years. Technical complications included implant fractures, connection-related and suprastructure-related complications. The cumulative incidence of implant fractures after 5 years was 0.4% (95% CI: 0.1 1.2%). After 5 years, the cumulative incidence of connection-related complications (screw loosening or fracture) was 7.3% and 14% for suprastructure related complications (veneer and framework fracture). CONCLUSION: Despite a high survival of FPDs, biological and technical complications are frequent. This, in turn, means that substantial amounts of chair time have to be accepted by the clinician following the incorporation of implant-supported FPDs. More studies with follow-up times of 10 and more years are needed as only few studies have described the long-term outcomes. PMID- 15533125 TI - A systematic review of the survival and complication rates of fixed partial dentures (FPDs) after an observation period of at least 5 years. II. Combined tooth--implant-supported FPDs. AB - OBJECTIVES: The objective of this systematic review was to assess the 5- and 10 year survival of combined tooth-implant-supported fixed partial dentures (FPDs) and the incidence of biological and technical complications. METHODS: An electronic MEDLINE search supplemented by manual searching was conducted to identify prospective and retrospective cohort studies on FPDs with a mean follow up time of at least 5 years. Patients had to have been examined clinically at the follow-up visit. Assessment of the identified studies and data abstraction was performed independently by two reviewers. Failure and complication rates were analyzed using random-effects Poisson regression models to obtain summary estimates of 5- and 10-year survival proportions. RESULTS: From a total of 3844 titles and 560 abstracts, 176 articles were selected for full-text analysis, and 13 studies met the inclusion criteria. Meta-analysis of these studies indicated an estimated survival of implants in combined tooth-implant-supported FPDs of 90.1% (95 percent confidence interval (95% CI): 82.4-94.5%) after 5 and 82.1% (95% CI: 55.8-93.6%) after 10 years. The survival rate of FPDs was 94.1% (95% CI: 90.2-96.5%) after 5 and 77.8% (95% CI: 66.4-85.7%) after 10 years of function. There was no significant difference in survival of tooth and implant abutments in combined tooth-implant FPDs. After an observation period of 5 years, 3.2% (95% CI: 1.5-7.2%) of the abutment teeth and 3.4% (95% CI: 2.2-5.3%) of the functionally loaded implants were lost. After 10 years, the corresponding proportions were 10.6% (95% CI: 3.5-23.1%) for the abutment teeth and 15.6% (95% CI: 6.5-29.5%) for the implants. After a 5 year observation period, intrusion was detected in 5.2% (95% CI: 2-13.3%) of the abutment teeth. Intrusion of abutment teeth were almost exclusively detected among non-rigid connections. CONCLUSION: Survival rates of both implants and reconstructions in combined tooth-implant supported FPDs were lower than those reported for solely implant-supported FPDs (Pjetursson et al. 2004). Hence, planning of prosthetic rehabilitation may preferentially include solely implant-supported FPDs. However, anatomical aspects, patient centered issues and risk assessments of the residual dentition may still justify combined tooth-implant-supported reconstructions. It was evident from the present search that tooth-implant-supported FPDs have not been studied to any great extent and hence, there is a definitive need for more longitudinal studies examining these reconstructions. PMID- 15533126 TI - A systematic review of the survival and complication rates of fixed partial dentures (FPDs) after an observation period of at least 5 years. AB - The present study was done to determine the long-term success and survival of fixed partial dentures (FPDs) and to evaluate the risks for failures due to specific biological and technical complications. A MEDLINE search (PubMed) from 1966 up to March 2004 was conducted, as well as hand searching of bibliographies from relevant articles. Nineteen studies from an initial yield of 3658 titles were finally selected and data were extracted independently by three reviewers. Prospective and retrospective cohort studies with a mean follow-up time of at least 5 years in which patients had been examined clinically at the follow-up visits were included in the meta-analysis. Publications only based on patients records, questionnaires or interviews were excluded. Survival of the FPDs was analyzed according to in situ and intact failure risks. Specific biological and technical complications such as caries, loss of vitality and periodontal disease recurrence as well as loss of retention, loss of vitality, tooth and material fractures were also analyzed. The 10-year probability of survival for fixed partial dentures was 89.1% (95% confidence interval (CI): 81-93.8%) while the probability of success was 71.1% (95% CI: 47.7-85.2%). The 10-year risk for caries and periodontitis leading to FPD loss was 2.6% and 0.7%, respectively. The 10-year risk for loss of retention was 6.4%, for abutment fracture 2.1% and for material fractures 3.2%. PMID- 15533127 TI - A systematic review of the survival and complication rates of fixed partial dentures (FPDs) after an observation period of at least 5 years. AB - OBJECTIVES: The objective of this systematic review was to assess the survival of cantilever fixed partial dentures (FPDs) and the incidence of biological and technical complications. METHODS: An electronic MEDLINE search supplemented by manual searching was conducted to identify prospective and retrospective cohort studies on FPDs with a mean follow-up time of at least 5 years. Patients had to have been examined clinically at the follow-up visit. Assessment of the identified studies and data abstraction was performed independently by two reviewers. Failure and complication rates were analyzed using random-effects Poisson regression to obtain summary estimates of 10-year survival proportions. RESULTS: From a yield of 3658 titles and 211 abstracts, 81 articles were selected for full-text analysis, finally resulting in 13 studies that met the inclusion criteria. Meta-analysis of these studies resulted in an estimated survival rate of cantilever FPDs of 81.8% (95 percent confidence interval (95% CI): 78.2-84.9%) and success rate (free of all complications) of 63% (95% CI: 54.7-70.2%) after 10 years. The most common biological complication was loss of pulp vitality (32.6%) followed by caries at abutment teeth (9.1%). After a 10-year observation period 2.6% of the FPDs were lost as a result of dental caries and 1% due to recurrent periodontitis. The most frequent technical complication was loss of retention (16.1%) followed by material fractures (5.9%). The cumulative incidence of fractures of abutment teeth was 2.9% and 2.4% of the FPDs were lost as a result of abutment tooth fracture after an observation period of 10 years. CONCLUSION: This systematic review on cantilever fixed partial dentures indicated that survival and success rates of cantilever fixed partial dentures were lower than those of conventional end-abutment supported FPDs described in a review by Tan et al. (2004) and biological and technical complications were frequent. PMID- 15533128 TI - Comparison of implant survival following sinus floor augmentation procedures with implants placed in pristine posterior maxillary bone: a systematic review. AB - Sinus augmentation is a commonly used procedure in implant dentistry. However, a general consensus on implant survival after this procedure is still lacking. The objective of this study was to systematically review implant survival following sinus augmentation procedures compared with conventional implant placement in the posterior maxilla. Following the production of a detailed protocol, screening and quality assessments of clinical trials were conducted in duplicate and independently. The search yielded 579 abstracts and 93 were selected for full text screening. Six publications (five studies) fulfilled all the inclusion criteria and were relevant to the study. Heterogeneity of the selected papers prevented meta-analysis. Implant survival ranged from 73% to 100% for non augmented sinuses and from 36% to 100% for augmented sinuses in patient-based data. From implant-based data, survival varied between 75% and 100% for both non augmented and augmented areas. Implant survival appears to show greater variability in grafted sinuses than in the posterior maxilla. However, prospective studies with larger patient numbers and control of confounding factors are urgently needed to provide definitive data on this important procedure. PMID- 15533129 TI - A systematic review of the influence of different titanium surfaces on proliferation, differentiation and protein synthesis of osteoblast-like MG63 cells. AB - OBJECTIVES: Titanium is the standard material for dental and orthopaedical implants. The good biocompatibility has been proven in many experimental and clinical investigations. Different titanium topographies were tested in vitro using different cell culture models. The aim of this systematic review was to evaluate and summarize the medical/dental literature to assess on which kind of titanium surface structure the osteoblast-like osteosarcoma cells MG63 show the best proliferation and differentiation rate, and the best protein synthesis. METHODS: A systematic search was carried out using different on-line databases (PubMed, Web of Science, Cochrane Library, International Poster Journal), supplemented by handsearch in selected journals and by examination of the bibliographies of the identified articles. Inclusion and exclusion criterias were applied when considering relevant articles. Studies which met the inclusion criteria were included and data extraction was undertaken by one reviewer. RESULTS: The search yielded 348 references. Nine articles referring to nine different studies were relevant to our question. Additionally 8 less relevant articles were identified. It was found that regularly textured surfaces of pure titanium with R(a) values (average roughness) of around 4 mum are well-accepted by MG63 cells. CONCLUSIONS: The surfaces and culture conditions vary widely. Therefore it is still difficult to recommend one particular surface. It seems that there are no differences in cell proliferation and differentiation on surfaces treated by blasting and etching. Standardization in fabrication and size of the different test surfaces as well as homogeneity in culture times and plating densities should be aspects for future research. PMID- 15533130 TI - Long-term implant survival in the grafted maxilla: results of a 12-year retrospective study. AB - OBJECTIVE: The aim of this study was to determine the long-term outcome of implant insertion in the augmented severely atrophied maxilla. STUDY DESIGN: Three hundred and twenty-four implants were inserted in 35 patients (eight males, 27 females, average age 57.6 years) in extremely atrophied maxillae after osteotomy and interposition of iliac crest bone. One hundred implants were installed in 12 patients simultaneously with the osteotomy and grafting; 224 implants were placed in 23 patients in a second procedure 6-12 months later. Implant parameters like osseointegration and peri-implant bone loss; peri-implant tissue parameters like bleeding, gingival and plaque index; and patients' satisfaction were evaluated. RESULTS: Of 324 implants, 29 (8.9%) were lost during the entire follow-up: 14 in six patients of the one-step and 15 in 11 patients of the two-step group. The overall input-output survival in 141.1 months was 91.1%. The overall 2-year failure-free fraction of implants was 95.5%; the 5-year failure-free fraction was 89.3%. In the one-step group, the 2 (5)-year failure free fraction was 95.9% (86.9%), and in the two-step group 95% (91.3%) (log-rank test P=0.57). Marginal peri-implant bone loss was 1.7+/-1.3 mm mesial and 1.8+/ 1.3 mm distal. CONCLUSION: Implant insertion after osteotomy and iliac bone grafting is a reliable operation method for the dental rehabilitation of the severely atrophied maxilla showing good long-term results. PMID- 15533131 TI - A clinical investigation of the morphological changes in the posterior mandible when implant-retained overdentures are used. AB - OBJECTIVES: The widespread use and broad acceptance of osseointegrated dental implants as a means of retaining and supporting mandibular overdentures provide potentially stable three-dimensional reference structures, which have been used to examine the morphological changes of the surface contours of the mandibular ridges distal to implants. Unlike traditional methods that tended to measure changes in ridge morphology and quantity radiographically, this prospective study investigated the changes in the sagittal surface contours of the posterior residual ridges over a 1-year period by an alternative, non-radiographic method using serial casts of residual ridges of a group of patients. MATERIAL AND METHODS: The residual ridges of the serial casts produced were examined using a profiling apparatus and computer software programs that allowed the orientation, digitised recording and measurement of the sagittal surface contours of the posterior residual ridges of mandibular casts in terms of area (cm(2)). RESULTS: Statistical analysis revealed that the detected reductive changes in the surface contour of the posterior residual ridges following the wearing of an implant retained mandibular overdenture over a 1-year period were highly significant (P<0.001). CONCLUSION: It is concluded that the methodology presented in this study can be used reliably as an alternative to radiographical examination for detecting changes that occur in the distal residual alveolar ridges of the implant containing edentulous mandible when implant-retained overdentures are used. Thus, the method presented may be used to contribute to a reduced exposure to ionising radiation for patients wearing implant-retained overdentures. PMID- 15533132 TI - The effects of naproxen sodium on specific parameters of L-strain cells over guided bone regeneration barrier materials. AB - There is strong evidence that nonsteroidal antiinflammatory drugs (NSAID) may exert a significant antiproliferative effect. This study evaluated the influence of NSAID on specific parameters of fibroblastic cells, in vitro, over two-guided bone regeneration (GBR) barrier materials. Fibroblast cells were cultured on bioabsorbable membrane made of collagen (Bio-Gide(R)- BG) and the most common nonresorbable membrane which is made of expanded polytetrafluoroethylene (ePTFE, Gore-Tex(R)- GT). Naproxen sodium (10 mM) was used as an analgesic drug. The fibroblast cells were cultured in vitro for 24 h and examined by scanning electron microscopy (SEM). Cells were cultured in the presence of (3)H-thymidine to study cell proliferation. And also cell numbers and viabilities were measured. The difference between the means for each group were analyzed for statistical significance by Kruskal-Wallis one-way ANOVA followed by post hoc comparisons using the Dunn statistical method. Of all the six groups, the control group stimulated DNA synthesis more than the others. With respect to cell numbers, there was statistically significant difference between the control group and naproxen planted BG membrane group. The interpretation of our SEM images is that these two barriers and naproxen seem to have had the least effect on cellular morphology. These data suggest naproxen have an inhibitory effect on stimulation of DNA synthesis, cell numbers and viabilities. And also lacking adherence of cells to the membranes may be due to the physical properties of the materials. PMID- 15533133 TI - Sinus floor augmentation with simultaneous placement of dental implants in the presence of platelet-rich plasma or recombinant human bone morphogenetic protein 7. AB - The aim of the present study was to evaluate the possible benefit of platelet rich plasma (PRP) in sinus grafting as compared with recombinant human bone morphogenetic protein-7 (rhBMP-7). For this purpose, we performed a bilateral sinus augmentation with anorganic bovine bone and simultaneous insertion of a titanium screw implant in five miniature pigs. Six hundred microliters of PRP and 15%-vol. autologous bone, which was collected with a trap during preparation of the implant recipient site, were added to the right sinus and 420 microl rhBMP-7 to the left sinus. A polychrome sequential labeling was performed. The animals were sacrificed 6 weeks after surgery. Undecalcified ground sections were evaluated by microradiography, digitized histomorphometry and under fluorescent light. The mean bone-implant contact using rhBMP-7 was 45.8% and 5.7% under PRP (P=0.002). The mean height of newly mineralized bone in the augmented area using rhBMP-7 amounted to 8.3 mm as opposed to 3.6 mm under PRP (P=0.013). Using PRP, the mean area of the newly formed bone was enhanced (51.3%) as compared with rhBMP-7 (33.1%); however, this difference was not statistically significant (P=0.081). In conclusion, under the selected experimental conditions the use of rhBMP-7 led to superior outcomes with regard to the osseointegration of dental implants and the height of new bone as compared with the use of PRP. PMID- 15533134 TI - Histomorphometry of human sinus floor augmentation using a porous beta-tricalcium phosphate: a prospective study. AB - Tricalcium phosphate (TCP) has been historically a well-accepted material for bone augmentation. We examined the use of a porous beta-TCP (100%) in a split mouth model for sinus floor augmentation. Five patients were treated bilaterally, receiving 1-2 mm-sized beta-TCP particles (Cerasorb) on one side (test side) and autologous chin bone particles on the other (control) side. Four other patients were treated with a unilateral sinus floor augmentation using 100% beta-TCP (no controls). Biopsies of the augmented sites were taken at 6 months. Histomorphometry measurements were carried out in order to quantify bone augmentation at test and control sides. The average bone volume (BV) formed in the augmented sinus at the control side was 41% (32-56%) and 17% (9-27%) in the test side when all nine patients were included (statistically significant, P=0.04). When only the five bilateral patients were included, mean BV of the test side was 19% (13-27%), which was also significantly different from the control side (P=0.009). Osteoid formation tended to be higher in the test side biopsies (1.3%) than in the controls (0.3%) (marginally significant, P=0.1), indicating ongoing bone formation in the TCP material. The amount of lamellar bone at the test side was less than half the amount in the control side, indicating that remodelling had only recently started in the TCP-augmented side. The resorption surface, however, did not differ significantly between the two sides. These histological results indicate that Cerasorb is an acceptable bone substitute material for augmentation of the maxillary sinus. Due to the osteoconductive, but not osteoinductive properties of this material, the rate of bone formation is somewhat delayed in comparison to autologous bone. PMID- 15533135 TI - Bone formation following sinus grafting with autogenous bone-derived cells and bovine bone mineral in minipigs: preliminary findings. AB - Bone formation in a sinus grafted with a cell-free scaffold requires the presence of local progenitor cells that differentiate into osteoblasts. The purpose of this study was to examine the effect of culture expanded autogenous bone-derived cells (ABC) with bovine bone mineral (BBM) on bone formation after single-stage sinus grafting in minipigs. Bone biopsies from the iliac crest were harvested 4 weeks prior to sinus grafting and ABC were culture expanded in vitro. The sinuses of five adult minipigs were grafted. In one sinus of each minipig the space between Schneider's membrane (SM) and the sinus wall was grafted with ABC (325,000 cells per sinus, on average) and BBM. In the other sinus, BBM alone was used. The animals were sacrificed after 12 weeks. One block of each grafted area was prepared by saw cutting and the amount of newly formed bone was analysed by micro-computed tomography (mu-CT). The addition of ABC to BBM significantly increased the amount of newly formed bone compared with BBM alone on mu-CT analysis (ABC+BBM: 29.86+/-6.45% vs. BBM: 22.51+/-7.28% (P=0.016)). Bone formation was increased near SM (ABC+BBM: 20.7+/-4.5% vs. BBM: 15.43+/-3.62% (P=0.009)) and in the middle zone of the grafting material (ABC+BBM: 31.63+/ 7.74% vs. BBM: 22.5+/-7.91% (P=0.001)), but not near the local host bone (ABC+BBM: 37.23+/-8.23% vs. BBM: 28.42+/-12.54% (P=0.15)). These preliminary findings indicate that supplementation of cell-free grafting material with culture expanded ABC can stimulate bone formation in areas with low bone-forming capacity. PMID- 15533136 TI - The use of porous calcium phosphate scaffolds with transforming growth factor beta 1 as an onlay bone graft substitute. AB - OBJECTIVES: Autogeneous bone grafting is regarded to be the golden standard for onlay grafts, but it requires a harvesting procedure and the remodeling pattern over time is unpredictable. New materials are constantly being sought to overcome these problems. An in vivo experiment was carried out to evaluate whether (1) porous calcium phosphate cement is a suitable biomaterial for onlay bone grafting, and (2) the addition of transforming growth factor beta 1 (TGF-beta1) accelerates de novo bone formation inside the cement porosity. MATERIAL AND METHODS: A carrier of porous calcium phosphate cement (Calcibon) was designed and 16 rats received one preshaped implant each. In 8 out of 16 implants 0.75 mug TGF beta1 was applied. The animals were killed after 4 weeks and the characteristics of tissue ingrowth into the onlay graft were evaluated. RESULTS: Histologic and quantitative histomorphometrical measurements demonstrated osteoid-like tissue formation in both experimental groups. The addition of TGF-beta1 did not induce significantly more osteoid-like tissue formation. On the other hand, in TGF-beta loaded implants, a higher number of pores contained an inflammatory infiltrate. CONCLUSION: This study indicated that porous calcium phosphate cement is a promising material for clinical situations where bone formation has to be supported. PMID- 15533137 TI - Relation between change of hearing and (modified) Amsterdam Inventory for Auditory Disability and Handicap Score. AB - This study investigates the test-retest distributions and the interval for true score change of the (modified) Amsterdam Inventory for Auditory Disability and Handicap [(m)AIAD], when the latter is used to measure the effect of an intervention. In a previous study the reliability and validity of the (m)AIAD in a cohort of hearing impaired patients were found to have satisfactory high values. In this prospective study, 66 patients underwent a tympanoplasty operation. Preoperatively and postoperatively pure tone audiometry was performed, and at the same time the subjective hearing ability was established by means of the (m)AIAD. The correlation between threshold change and score change was 0.35 (Pearson's r). Scores on the (m)AIAD had to change by at least 16 to be qualified as a true change. For only nine of 66 subjects this criterion was fulfilled. No clear relation exists, except for these nine subjects, between threshold change and score change in this patient population. The study also shows that disability questionnaires have their limitations, when using them to measure the result of a medical intervention in an individual patient. PMID- 15533138 TI - A retrospective review of stapes surgery following introduction of 'reversal of the steps' technique. AB - A retrospective study of 73 consecutive stapedotomies is reported. The technique of reversal of steps was achieved in 59% of cases. Closure of the air-bone gap to within 10 dB was achieved in 84% of the reversal group, and 81% of the standard technique group, with no statistically significant difference between the two groups. PMID- 15533139 TI - Advances in genetic manipulations in the treatment of hearing disorders. AB - Gene transfer into the inner ear is an attractive technology for clinical applications. It offers the hope of preventing, arresting, reversing or curing vestibular or hearing disorders caused by hereditary diseases or environmental insults. There are currently very few treatment options for vestibular disorders and sensorineural hearing loss, and therefore, it is important to investigate and develop new technologies for inner ear disease. In utero gene transfer may indeed be an option, but this technology will have many technical and ethical issues that remain to be overcome. PMID- 15533140 TI - Keratosis obturans and external ear canal cholesteatoma: how and why we should distinguish between these conditions. AB - Keratosis obturans and external ear canal cholesteatomas have been considered as separate entities for the last 20 years, after being regarded as variations of the same disease for at least 87 years. While both disorders are distinct, they do have some overlapping characteristics which may make it difficult to reach a definite diagnosis. This review explores the diagnostic dilemmas which may arise, and discusses the classification, aetiology, pathogenesis and management of these conditions. We concur that external ear canal cholesteatoma and keratosis obturans are different conditions and conclude that the presence of osteonecrosis and focal overlying epithelial loss are the most reliable features favouring the diagnosis of external ear canal cholesteatoma over keratosis obturans. Furthermore, whilst keratosis obturans can be managed successfully by regular aural toilet, external ear canal cholesteatoma may require surgical intervention depending on the extent of the disease. PMID- 15533141 TI - 'Moffett's' solution: a review of the evidence and scientific basis for the topical preparation of the nose. AB - General anaesthesia, often causes a widespread vasodilation, producing a hyperaemic nasal mucosa, hence the need for a topical application to decongest the nose and reduce the nasal blood flow to optimize the operative field. The use of a combination of cocaine, sodium bicarbonate and adrenaline given the eponymous title of 'Moffett's Solution' is standard practice in many rhinological procedures to provide local anaesthesia, vasoconstriction and decongestion. We discuss each component of 'Moffett's' reviewing the science and evidence behind its usage and the huge variation in the techniques of its application. PMID- 15533142 TI - Exostoses of the external auditory canal: a long-term follow-up study of surgical treatment. AB - To determine the postoperative incidence, extent and recurrence rate of exostoses of the external auditory canal in a cohort of patients involved in different water sports. A cross-sectional study of 31 patients (46 ears), with exostoses treated by surgery in the Royal Cornwall Hospital between 1980 and 1999. A questionnaire was used to obtain information about the type of water exposure pre and postoperatively. The extent of recurrent stenosis was assessed. The mean postoperative time interval was 10 years (sd = 4.5 years). The degree of stenosis was assessed as: minimal (<30%) in 42.6%, moderate (30-60%) in 31% and severe (>60%) in 25% of ears. The Cox regression model was used to identify factors associated with a reduction in the recurrence rate of stenosis. The use of ear plugs was highly significant (P = 0.015), as was the age of the patient at the time of operation (P = 0.004), i.e. the older the patient, the faster recurrent disease developed. There was no evidence to show that either the type or seasonal pattern of water sport activity influenced recurrence of the disease postoperatively, although preoperatively, the stenosis was more marked in association with surfing and sailing. Exostoses developed faster preoperatively in those who were in the water all year round rather than just the summer months. Of five patients who stopped water sport activity completely after surgery, four of them developed significant recurrent exostoses (>50% stenosis). PMID- 15533143 TI - Does ENT outpatient review at 1-week post ventilation tube insertion improve outcome at 1 month in paediatric patients? AB - The aims of this study were to see if ENT review at 1 week following ventilation tube insertion in paediatric patients reduced the complication rate and the number of consultations with the general practitioner in the first postoperative month. Sixty-six children were prospectively randomized into two follow-up groups: group A was reviewed at 1 week and 1 month and group B at 1 month only. There was no statistically significant difference in postoperative otorrhoea (P = 0.59), ventilation tube patency (P = 0.44), ventilation tube extrusion rate (P = 0.64) and number of consultations with the general practitioners (GP) for postoperative otorrhoea (P = 0.19) between the two groups at 1 month. In conclusion, there seems to be no advantage with early postventilation tube insertion review as it does not reduce complications or decrease the GP consultation rate. PMID- 15533144 TI - The use of perioperative Sofradex eardrops in preventing tympanostomy tube blockage: a prospective double-blinded randomized-controlled trial. AB - Around 11-12% of tympanostomy tubes are reported to become blocked by middle ear secretions or blood immediately following surgery, and so no longer function. Many otologists routinely instil an antibiotic and steroid-containing solution at the time of surgery in the belief that this may reduce this complication. The aim of the study was to investigate the efficacy of instilling the antibiotic and steroid-containing solution Sofradex at the time of grommet insertion in preventing grommet blockage. Double-blind randomized-controlled trial, comparing rates of grommet blockage in ears treated with Sofradex drops against control (no drops) in patients undergoing bilateral grommet insertion. Sixty-one pairs of results were obtained. There was a significant difference between the rates of grommet blockage in the two groups. Grommets with Sofradex drops instilled perioperatively were nine times less likely to be blocked than controls [1.6%versus 13.1%, odds ratio (Sofradex/control) = 9.06, 95% confidence interval (CI): 1.04-78.82, P = 0.05]. There was no association between grommet blockage and perioperative bleeding or the nature and presence of middle ear secretions. Sofradex eardrops are effective in reducing the rate of grommet blockage when instilled perioperatively. PMID- 15533145 TI - Otic drops used to clear a blocked grommet: an in vitro prospective randomized controlled study with blinded assessment. AB - Clinicians often prescribe otic drops anecdotally to try and clear grommets blocked with blood. We carried out an in vitro double-blind randomized controlled study comparing the efficacy of sodium bicarbonate, Locorten Vioform and olive oil drops in clearing Shah grommets placed in 'artificial ears' and blocked with blood in a standardized fashion. There were 33 grommets in each group, and drops were inserted three times a day for 7 days. Olive oil drops cleared 17 of 33 (51.51%), Locorten Vioform cleared one of 33 (3%) and sodium bicarbonate cleared zero of 33 (0%) blocked grommets. Statistical comparison between pairs indicates that olive oil was significantly better than both Locorten Vioform (P < 0.001) and sodium bicarbonate drops (P < 0.001) at clearing grommets blocked with blood. PMID- 15533146 TI - Outcomes of myringoplasty in Australian Aboriginal children and factors associated with success: a prospective case series. AB - The objective of this study was to assess the outcomes of myringoplasties in Aboriginal children and to identify factors associated with a successful outcome with the use of prospective case series from primary health care clinics and hospitals in four rural and remote regions of Western Australia. All 58 Aboriginal children, aged 5-15 years, who underwent 78 myringoplasties between 1 January 2000 and 30 June 2001 were included in the study. Complete postoperative (post-op) follow-up was achieved following 78% of myringoplasties. The main outcome measures were (a) success, i.e. an intact tympanic membrane and normal hearing six or more months post-op in the operated ear, (b) closure of the perforation, (c) Post-op hearing improvement. Forty-nine per cent of myringoplasties were successful, 72% resulted in closure or reduction in the size of the perforation and 51% resulted in hearing improvement. After controlling for age, sex, clustering and number of previous myringoplasties, no association was observed between success or hearing improvement and perforation size, or the presence of serous aural discharge at the time of surgery. Myringoplasty resulted in hearing improvement and/or perforation closure in a significant proportion of children. Thus, primary school-aged Aboriginal children in whom conservative management of chronic suppurative otitis media has been unsuccessful should have access to myringoplasty because of the positive impact on their socialization, language and learning that results from improved hearing. PMID- 15533147 TI - Tympanoplasty - reporting hearing results and 'hearing objective'. AB - When reporting the results of tympanoplasty, the postoperative air-bone gap (ABG) presented in 10 dB bins, ABG closure and air conduction threshold gain are commonly reported indicators of tympanoplasty outcome. When tympanoplasty is performed, the reconstruction aims either to improve hearing threshold or to maintain satisfactory thresholds, that is, the surgical intention is either for 'hearing gain' or for 'hearing preservation'. This review of the early results of tympanoplasty examines whether classifying surgery as either for hearing gain or for hearing preservation influences the reported results. Closure of the ABG to within 20 dB was achieved in 72-94% of cases, the average postoperative ABG was between 13.1 and 17.1 dB with the postoperative air conduction threshold being between 27.4 and 33.5 dB. These figures were similar for both hearing preservation and hearing gain procedures. However air conduction threshold gain was significantly greater for the 'hearing gain' group (17 dB versus 0 dB) and was reduced to 8 dB by combining the two groups. Overall, indicating whether surgery attempted hearing preservation or hearing gain did not significantly alter the parameters used for reporting tympanoplasty outcome. PMID- 15533148 TI - Minimally invasive approach and fixation of cochlear and middle ear implants. AB - One of the conventional surgical approaches for cochlear implantation is a retro auricular incision with a posterior-inferiorly based skin and subcutaneous tissue flap and a superiorly based periosteal flap. The obvious advantage is an open operating field but the disadvantages are a large wound and a lengthy operating time. It may also result in more wound-related complications. To overcome these disadvantages, we have developed a minimally invasive technique that includes a small retro-auricular single layer incision. We have used a metal bridge beneath the posterior flap to increase accessibility when creating a recess for the implant. A novel technique is used to place the securing suture deep to the flap. This technique has been used in 49 paediatric and adult patients, and there have been no wound-related complications. Although this technique was initially designed for the CLARION CII implant, it has been used to place and secure the new CLARION HiRes 90 K, the Nucleus device, the MEDEL device and the Vibrant Soundbridge. PMID- 15533149 TI - Quality of life following microsurgery, radiosurgery and conservative management for unilateral vestibular schwannoma. AB - The best way to manage small and medium-sized vestibular schwannomas is currently a matter of heated debate. As these tumours are not immediately life-threatening, patients are invariably concerned about how management would affect their quality of life. Until now, no study has compared the three treatment modalities in terms of physical, psychological and social wellbeing. This study is based on a retrospective database analysis and postal questionnaire survey of unilateral vestibular schwannoma patients who had either been managed conservatively, or treated with microsurgery or radiosurgery. The results showed that: quality of life (measured by the Glasgow Benefit Inventory) deteriorated after microsurgery, particularly for small tumours; conservative management did not lead to a change in quality of life, and there was a trend towards poorer quality of life following radiosurgery. The findings suggest that a conservative management approach may be more appropriate for small tumours, and that patients who are due to undergo microsurgery or radiosurgery may benefit from counselling about the potential impact of treatment on quality of life. PMID- 15533150 TI - Tinnitus and psychological morbidity: a cross-sectional study to investigate psychological morbidity in tinnitus patients and its relationship with severity of symptoms and illness perceptions. AB - This study investigates the relationship between tinnitus and psychological morbidity using a cross-sectional survey design from patients of an outpatient audiology department in a typical District General Hospital. The results suggest that a significant number of tinnitus patients suffer from psychological problems both during and more unfortunately after routine treatment for their tinnitus. Anxiety was found to be the main psychological problem. In conclusion, this study suggests that, while further research is needed, many tinnitus patients require psychological treatment and this has implications for the staffing of audiology units in District General Hospitals. PMID- 15533151 TI - Reduction in noise-induced temporary threshold shift in humans following oral magnesium intake. AB - A previous study demonstrated the prophylactic effects of magnesium on noise induced permanent threshold shift in humans. For the first time, this study explores the effects of magnesium on temporary threshold shift in 20 human subjects, all men (16-37 years, mean age 21 years). The study was conducted in a double-blind manner on the same subjects tested in three different phases (placebo, magnesium, no-drug). The cochlear changes were assessed using both behavioural (audiograms) and objective (otoacoustic emission) measures. Blood samples were collected at the beginning and end of each phase. Amongst subjects and phases, magnesium intake was associated with significantly lower temporary threshold shift, compared with the other two phases, which were reflected both by the behavioural and cochlear measures. A correlation was found between the blood magnesium levels and temporary threshold shift reduction. No side-effects were associated with the oral ingestion of the magnesium. Magnesium provides significant protection against temporary threshold shift, complementing the previous permanent threshold shift human study. Both human noise-induced hearing loss studies introduced a novel, biological, natural agent for prevention and possible treatment of noise-induced cochlear damage in humans. PMID- 15533152 TI - Numerical simulation of air temperature and airflow patterns in the human nose during expiration. AB - Recovery of heat and water during expiration is an important but not yet fully understood function of the nose. The presented study investigated cooling of the expiratory air for heat recovery within the human nose applying numerical simulation. A numerical simulation in a bilateral three-dimensional model of the human nose based on computed tomography was employed. Temperature distribution and airflow patterns during expiration were displayed. Cooling of the expiratory air primarily takes place in the areas of inferior and middle turbinate. Areas of the highest decrease in temperature are characterized by turbulent airflow with vortices of low velocity. Numerical results showed good concordance with experimental in vivo temperature measurements. Heating of inspired air not only depends on inspiration but also on expiration. Cooling the warm expiratory air may be regarded as an important factor for heat recovery. Furthermore, the results demonstrate the close relation between heat exchange and airflow patterns. PMID- 15533153 TI - The smell map: commonality of odour perception confirmed. AB - We have examined the repeatability of a perceptual space, or smell map, initially developed in a sample of 20 people. A total of 110 normosmic people performed similarity ratings for pairs of the 11 commonly encountered odours used in the previous study. Multidimensional scaling was again used to analyse the relationships between odours. Some individual plots and an archetype, generated using the data from all participants, produced results similar to the previous study, confirming the ability to obtain reproducible results using a different tester and different participants. Exploration of the source space, that is, the spatial distribution of the individual plots relative to each other, suggests that the current model explains about 50% of the variation between subjects for the two-dimensional solution and 57% for the three-dimensional solution. PMID- 15533154 TI - Airborne environmental pollutant concentration and hospital epistaxis presentation: a 5-year review. AB - To investigate the hypothesized relationship between various daily atmospheric pollutant concentrations and hospital epistaxis presentation. A retrospective analysis of all presentations of non-traumatic epistaxis to our institution was performed, and the results were cross-referenced with London air quality data supplied by the National Air Quality Data Archive at the National Environmental Technology Centre (NETCEN) at the multi-centre tertiary referral ENT Department. An analysis of epistaxis presentation to St George's Hospital during the 5-year period January 1997-2002 was made from the patient administration system and accident and emergency records. The study includes 1373 emergency patients after traumatic and iatrogenic epistaxis were excluded. Factors measured were atmospheric ozone (O(3)), carbon monoxide (CO), sulphur dioxide (SO(2)), nitrogen dioxide (NO(2)) and particulate matter less than 10 mum in diameter (PM(10)). Hospital epistaxis presentation and atmospheric pollutant concentration were documented for each day of the 5-year period, weekly change in both parameters was recorded and statistical analysis was performed. The statistical methods used are linear correlation using Pearson's coefficient. Increased hospital epistaxis presentation was strongly associated with increased concentration of airborne particulate matter (r = 0.289, P < 0.001; significant if P < 0.05) and less strongly associated with increased concentrations of atmospheric O(3) (r = 0.150, P = 0.019; significant if P < 0.05). There was no association between epistaxis attendance and atmospheric concentrations of CO, NO(2) and SO(2) in this study. The concentrations of local airborne particulate matter and atmospheric O(3) in the days preceding hospital attendance for epistaxis are relevant when considering the associations of the episode. This further increases understanding of the pathophysiology of spontaneous epistaxis. PMID- 15533155 TI - Nasal congestion: mechanisms, measurement and medications. Core information for the clinician. AB - The article provides the clinician with core information about the aetiology and treatment of nasal congestion and discusses nasal blood vessels, autonomic nerves, nasal cycle, effects of posture, subjective sensation of congestion, objective assessments of congestion and medical and surgical treatments. PMID- 15533156 TI - Which solvent for olfactory testing? AB - The physical properties of any carrier can deteriorate over time and thus alter the results in any olfactory test. The aim of this study was to evaluate clinically potential solvents as a clean odourless carrier for olfactory testing. Sweet almond oil, pure coconut oil, pure peach kernel oil, dipropylene glycol, monopropylene glycol, mineral oil and silicone oil were studied. The experimentation was conducted in two parts. First, an olfactory device was used to conduct air through the solvents on a weekly basis using a cohort of six volunteers to assess the perceived odour of each solvent at weekly intervals. Secondly a cross-reference test was performed using small bottled solutions of phenylethyl-alcohol and 1-butanol in 10-fold dilutions to compare any perceived difference in concentrations over a period of 8 weeks. We concluded that mineral oil is the most suitable carrier for the purpose of olfactory testing, possessing many desirable characteristics of an olfactory solvent, and that silicone oil may provide a suitable alternative for odorants with which it is miscible. PMID- 15533157 TI - A comparison of aesthetic proportions between the Oriental and Caucasian nose. AB - Differences in aesthetics between the Oriental and Caucasian nose were examined from a cohort of 118 healthy volunteers of which 61 had an Oriental and 57 had a Caucasian racial origin. The oriental nose projected less from the face, and was broader at the intercanthal level (P < 0.001) and the alar base (P < 0.001), but not at the bony base. The oriental nose projected less at all levels: nasion projection (P < 0.001) and tip projection (P < 0.001). The naso-labial angles for the Orientals in this study exhibited a wider range of variation than the Caucasians, with the oriental male exhibiting the most acute angle (average 86.2 degrees ), because of the forward angulation of the upper lip. Many of the average aesthetic parameters obtained from these two cohorts of healthy subjects did not conform to the published aesthetic standards, set out as the ideal goals for rhinoplasty. PMID- 15533158 TI - Vascular endothelial growth factor in nasal polyps: a comparison of asthmatic and non-asthmatic patients. AB - The cause of nasal polyps remains unknown, although there is a well-recognized clinical association between nasal polyposis and asthma. The characteristic histological features of nasal polyps include large quantities of extracellular fluid. Vascular endothelial growth factor (VEGF) is a potent mediator of angiogenesis and vascular permeability. This study aimed to compare expression of VEGF in nasal polyps from patients with asthma and those with no apparent respiratory disease. Twenty-four asthmatic and 35 non-asthmatic patients were studied using immunohistochemistry for VEGF. VEGF expression was identified in endothelial, inflammatory and epithelial cells. There was significantly greater endothelial expression of VEGF in asthmatic patients (P < 0.05). Greater epithelial expression was observed in asthmatic patients but this did not reach statistical significance (P = 0.07). There was no difference in the density of inflammatory cells expressing VEGF. Differences between the two groups may reflect differences in disease severity or in the nature of the inflammatory process. PMID- 15533159 TI - Increase of the apnoea-hypopnoea index after uvulopalatopharyngoplasty: analysis of failure. AB - From 70 patients who had uvulopalatopharyngoplasty (UPPP) operation and a pre- and postoperative sleep registration, we could retrospectively determine the failures and the correlation between variables such as age, gender, body mass index (BMI), earlier or concomitant tonsillectomy, unilevel (uvula-palate-tonsil) or multilevel (base of tongue as well) obstruction during sleep endoscopy and treatment outcome. From 70 patients, the preoperative sleep registration classified 15 social unacceptable snorers and 55 obstructive sleep apnoea syndrome (OSAS) patients. In this study we focused on the OSAS patients. From the 55 OSAS patients, 32 were classified as successful after UPPP, because they had a decreased apnoea-hypopnoea index (AHI) after surgery ( 0.56) as between the level of obstruction(s) (P > 0.24). For earlier or concomitant tonsillectomy we found a statistically difference (P > 0.039), but a very small number in the high failure group (n = 8). We conclude that although sleep endoscopy adds to better patient selection and better results, paradoxically, the finding of obstruction on palate-uvula level during sleep endoscopy can still give UPPP failures. PMID- 15533160 TI - Velopharyngeal insufficiency following adenoidectomy. AB - Velopharyngeal insufficiency (VPI) is a well recognized but rare complication of adenoidectomy. Twenty children with this condition were seen and assessed at Great Ormond Street Hospital between 1993 and 2000. The commonest aetiology was occult submucous cleft palate (n = 5) but there was a wide range of other causes. Two children with severe behavioural disorders and normal palates developed mild symptoms, an aetiology not previously reported. Only two children had a classical submucous cleft palate. Nine children required surgical intervention and three revision procedures. Of the 15 treated children for whom follow-up data was available, 13 regained normal or near-normal speech. Many cases of postadenoidectomy VPI was not foreseeable. Following referral to a specialist cleft unit, normal or near-normal speech can be achieved in the majority with a combination of surgery and speech therapy. PMID- 15533161 TI - Why do we have a uvula?: literature review and a new theory. AB - From ancient times up to the present day many different functions and conditions have been attributed to the uvula, many speculative and some with a more scientific basis. The uvula has been shown to have the ability to produce and secrete large quantities of thin saliva. A common complication of surgery involving removal of the uvula is pharyngeal dryness. We have observed that on phonation and swallowing the uvula swings back and forth in the oropharynx. We present a review of the literature on the uvula and propose a theory that the uvula bastes the throat and thereby helps keep it moist and well lubricated. PMID- 15533162 TI - Naturally occurring fruit juices dislodge meat bolus obstruction in vitro. AB - Oesophageal meat bolus obstruction is often removed endoscopically/surgically with attendant morbidity and mortality. Anecdotal evidence suggests that fruit juices' enzymes may dissolve meat boluses. To assess this, an in vitro study was carried out. Cooked chicken was tightly packed between 10 and 8 mL markers of 80, 10 mL syringes. Two millilitres of fresh juice from lemons, papaya, kiwi fruits, pineapples, frozen and then thawed pineapple juice, processed pineapple juice, Coca-Cola and saliva were added to 10 syringes each. By 4 h, the meat boluses significantly moved in the pineapple juice both fresh and frozen and kiwi juice groups (P < 0.001), little movement in the Coca-Cola group (P < 0.05), and no movement in the saliva, processed pineapple juice, lemon juice and papaya groups. In conclusion, pineapples and kiwi fruits contain enzymes which can dislodge meat boluses. Freezing does not denature the enzyme and provides a convenient way of storing the juice. PMID- 15533163 TI - Molecular markers in dysplasia of the larynx: expression of cyclin-dependent kinase inhibitors p21, p27 and p53 tumour suppressor gene in predicting cancer risk. AB - Premalignant conditions affect the larynx. Dysplasia can progress in severity resulting in cancer depending on many clinical, pathological and molecular factors. The purpose of this study was to examine the expression of the p21 and p27 cyclin-dependent kinase inhibitors and p53 tumour suppressor gene in dysplasia of the larynx. A total of 114 cases of untreated dysplasia were selected from the archives of the University of Newcastle. p21, p27 and p53 immunohistochemistry was performed and the cases followed up. Twenty-eight dysplasias (24%) subsequently developed into cancers. Expression of the molecular factors studied was not associated with cancer progression. p53 expression was associated with smoking (P = 0.005). In contrast, grade of dysplasia was significantly associated with cancer risk (odds ratio 6.7; P = 0.0001). The majority (75%) of cancers were detected within 12 months of dysplasia being diagnosed. PMID- 15533164 TI - Defining the criteria for radioresistant laryngeal cancer. AB - Survival rates for head and neck cancer are comparatively poor, typically 40% at 5 years. Radiotherapy is one of the most common modalities used to cure early stage cancers. It has the advantage in that it preserves anatomical structure and function. However, treatment failures do occur necessitating salvage surgery if a cure is to be achieved. A universally accepted definition of radioresistant cancer does not exist. Second primaries and occult metastasis are common in head and neck cancer, and can be confused with true radioresistant tumours. We suggest a strict definition for radioresistant laryngeal cancer and characterize 66 radioresistant tumours stage matched to 66 radiosensitive tumours. It was not possible to differentiate the radioresistant group from the radiosensitive groups using tumour differentiation. By using an agreed set of criteria defining radioresistant head and neck tumours, researchers will be better able to investigate molecular and cellular markers of radioresistance. PMID- 15533165 TI - The role of glottal gap in predicting aspiration in patients with unilateral vocal paralysis. AB - Aspiration has been frequently noted among patients suffering from vocal-fold paralysis. The association between a defect of glottal closure and aspiration in patients with unilateral vocal paralysis appears to have been rarely reported. The current study reviews collected data consisting of videotaped flexible nasoendoscopy and stroboscopy for patients with a unilateral vocal-fold paralysis. Mean normalized glottal gap areas for patients suffering vocal paralysis with associated aspiration are significant greater than that for the non-aspiration group. Subsequent to the surgical correction of the glottal gap, all patients recovered well from their earlier aspiration. The investigation of objectively derived data revealed that poor airway protection due to incomplete closure of the vocal fold was the major cause of aspiration for patients suffering unilateral vocal-fold paralysis. Intracordal autologous fat injection can successfully improve the breath control and eliminate aspiration in patients with vocal-fold paralysis. PMID- 15533166 TI - 3-D optical coherence tomography of the laryngeal mucosa. AB - Laryngeal carcinoma is one of the commonest primary head and neck malignancy and the need for early identification is very important for successful treatment. Outpatient fibreoptic examination of the larynx is unreliable in differentiating benign, pre-malignant and malignant lesions, and therefore surgeons have to rely on biopsies for a definitive diagnosis. This is an invasive procedure requiring general anaesthesia and may have a detrimental effect on the patient's voice. Conventional imaging modalities (ultrasound, computed tomography and magnetic resonance imaging) have a limited resolution and hence cannot give sufficient information on the extent or nature of laryngeal lesions. The aim of our study is to investigate the feasibility of optical coherence tomography (OCT) in imaging the normal larynx, to lay the foundations for an investigation of its ability to differentiate between benign and malignant disease. Ten tissue specimens from normal larynges were imaged with an 850 nm OCT system that was capable of providing both B-scan (longitudinal or cross-section) images as well as C-scan (en-face or images at constant depth). The en-face OCT mode allowed us to reconstruct 3-D OCT images of the tissue examined. Imaged specimens were processed with standard histopathological techniques and sectioned in the plane of the B-scan OCT images. Haematoxylin-eosin stained specimens were compared with the OCT images thus collected. Preliminary results showed good correlation between OCT images and histology sections in normal tissue. PMID- 15533167 TI - A blinded in vitro comparison of five sets of bipolar diathermy forceps. AB - The objective of this blinded randomized trial was to compare five sets of bipolar forceps, both single use and reusable, and discuss their potential role in postoperative complications. An animal tissue experiment was set up with standardized parameters. Results indicate larger tips give larger burns. We also found that some forceps gave suboptimal burns. PMID- 15533168 TI - Guidelines for the management of periorbital cellulitis/abscess. AB - Orbital and periorbital swelling secondary to acute sinusitis is relatively common. Distinguishing between preseptal and orbital involvement can be difficult based on clinical observations alone. A delay in diagnosis and appropriate treatment may result in serious complications including blindness. The literature is reviewed and simple guidelines suggested to manage such patients, including their initial assessment, investigations with particular reference to the indications and timing of CT scanning, antimicrobial therapy and the type of surgery. These guidelines have been approved across disciplines in our unit and trialled successfully for 2 years. PMID- 15533169 TI - Simultaneous radio- and chemotherapy for squamous cell carcinoma of the head and neck in daily clinical practice: 5 years experience in a University Hospital. AB - Several randomized studies and meta-analyses have shown that simultaneous radio- and chemotherapy prolongs survival in patients with unresectable squamous cell carcinoma of the head and neck as compared with conventional radiotherapy. We assessed the feasibility and effectiveness of simultaneous radiotherapy (35 x 2 Gy) and chemotherapy [cisplatinum 100 mg/m(2) or carboplatin (AUC 6) on days 1, 22 and 43] in daily clinical practice in a cohort of 87 patients treated at our institute between 1998 and 2002. Eighty patients completed radiotherapy according to schedule. Eighty patients received two courses of chemotherapy and 50 patients three courses. Nephrotoxity, bone marrow suppression and ototoxicity were the most frequent side-effects. Median weight loss was 8.5%. Median survival was 15 months and 44% of the patients were alive at 2 years. Patients receiving three courses of chemotherapy had a better survival than patients receiving two or less courses. Treatment with simultaneous radio- and chemotherapy for advanced head and neck cancer is a demanding, but feasible treatment in daily clinical practice. Survival seems to be comparable with the results achieved in patients selected for clinical trials. PMID- 15533170 TI - Management of serum calcium levels in post-thyroidectomy patients. AB - Otolaryngologists are increasingly performing thyroid surgery and are responsible for optimising parathyroid function postoperatively. The aim of this study was to establish an evidence-based algorithm for the management of postoperative calcium levels and to try and answer some relevant postoperative management issues. These include the following: (1) What is the risk of hypocalcaemia both temporary and permanent? (2) When should calcium replacement be commenced and in what form? (3) What is the best method for calcium replacement without suppressing a recovering parathyroid? (4) How to identify recovering parathyroid function if the patient is already on calcium replacement? A systematic review of the literature was carried out supported by a retrospective analysis of postoperative calcium levels obtained from clinical records of patients undergoing thyroidectomy under our care (n = 167). We present an evidence based, user-friendly algorithm for the management of the serum calcium in patients undergoing thyroid surgery. PMID- 15533171 TI - Percutaneous endoscopic gastrostomy (PEG) - a long-term follow-up study in head and neck cancer patients. AB - Many patients with head and neck cancer experience problems related to swallowing. A retrospective study of 156 consecutive patients who received a percutaneous endoscopic gastrostomy (PEG) at a teaching hospital is presented. The results showed that 42% had complications. Fatal complications were seen in connection with PEG tube placement, but severe and minor complications could occur much later. The method of PEG tube insertion did not affect the complication rates. The spectrum of observed complications is different to that reported earlier, suggesting that the learning curve of surgeons under training could have influenced the outcome. It may be concluded that for a very sick patient a theoretically easy surgical procedure could turn into a potentially dangerous operation. It is important to select suitable candidates for a PEG. Head and neck cancer patients with a PEG need special attention in connection with the PEG tube placement and also in a long perspective, e.g. by follow-up at a nurse-led outpatient clinic. PMID- 15533172 TI - Is cochlear outer hair cell function affected by mobile telephone radiation? AB - Mobile telephones emit high-frequency pulsed electromagnetic fields (PEMF). These are known to have measurable biological effects, and possible effects on the auditory system. Otoacoustic emissions give an indication of the functional state of the auditory system. Otoacoustics are known to be highly specific for the individual when the test pulse is identical. In this way, subtle changes in the ear can be detected. We investigated whether there is a measurable effect on Otoacoustic emissions from PEMF radiation. A total of 12 volunteers were recruited who had normal hearing; confirmed by pure tone audiometry. An Otoacoustic emission trace was obtained. The test subjects were exposed to a mobile telephone that was placed over the test ears mastoid process. The subjects had Otoacoustic emissions measured without the telephone and again on receive and transmit. There was no change in the trace signature during the test. There was no statistically significant change in the trace figures. This would indicate that PEMF from commonly available hand held mobile telephones have no measurable effect on the outer hair cell function during the time of use. PMID- 15533173 TI - How we do it: management of tracheobronchial foreign bodies in children. AB - KEY POINTS: Foreign body aspiration remains an important pathology in children. Management of airway foreign bodies must be undertaken in specialized units with experienced staff and known protocols. Development and improvement of rigid bronchoscopes, with advances in anaesthesia have increased the efficacy and the safety of removing endotracheal foreign bodies. A video monitor is advantageous in coordinating the procedure and reducing the stress of the team. Utilization of Fogarty's catheters can be beneficial. PMID- 15533174 TI - How we do it: a practical approach to Foley catheter posterior nasal packing. AB - KEY POINTS: There are various effective treatment options for refractory posterior epistaxis including endoscopic diathermy or endoscopic artery ligation. If these treatment options are not immediately available, posterior nasal packing with Foley catheter is an effective and rapid non-surgical treatment alternative. A systematic approach is presented. The principles and mechanisms of action of the posterior nasal packing are explained. Nevertheless, there are morbidities involved and posterior packing can be painful and uncomfortable for the patient. Marking and modifying the catheter will help one inflate the balloon in the correct place and avoid the problem of nasal alar pressure necrosis. PMID- 15533178 TI - From information on quality to quality information. PMID- 15533179 TI - Physical and sensory functioning over time and satisfaction with care: the implications of getting better or getting worse. AB - OBJECTIVE: To determine whether those persons whose sensory or physical functioning improved or worsened over one year are more or less satisfied with their health care. DATA SOURCES: 1996 and 1997 Medicare Current Beneficiary Survey (MCBS). STUDY DESIGN: The MCBS surveys a nationally representative, longitudinal panel of Medicare beneficiaries about their sociodemographic attributes; vision, hearing, and various mobility functions; and 10 items representing satisfaction with and access to health care. Using multivariable logistic regression and controlling for sociodemographic factors, we computed adjusted odds ratios of dissatisfaction with care, examining the effects of changes in sensory or physical functioning. Analyses accounted for MCBS sampling weights. DATA EXTRACTION METHODS: We identified 9,974 community-dwelling respondents, 18 years old and over who answered the 1996 and 1997 MCBS. We assessed five categories of sensory or physical functioning (vision; hearing; walking; reaching overhead; and grasping and writing) and compared 1996 and 1997 responses to identify those whose functioning improved or worsened. PRINCIPAL FINDINGS: Worsened functioning was strongly associated with older age, low income, and low educational attainment. Improved functioning was rarely significantly associated with satisfaction; an exception involved substantially lower rates of dissatisfaction with "ease and convenience" of getting to physicians. Worsened functioning was often statistically significantly associated with dissatisfaction, always with adjusted odds ratios >1.0. Across all five functional categories, persons whose functioning worsened displayed significantly greater dissatisfaction with overall quality, ease, and costs or care. CONCLUSIONS: Persons whose functioning improved rarely reported better satisfaction than did those whose functioning did not improve, while those whose functioning worsened expressed more systematic reservations about their care. PMID- 15533180 TI - The impact of diabetes on workforce participation: results from a national household sample. AB - OBJECTIVE: Diabetes is a highly prevalent condition that results in substantial morbidity and premature mortality. We investigated how diabetes-associated mortality, disability, early retirement, and work absenteeism impacts workforce participation. DATA SOURCE: We used the Health and Retirement Study (HRS), a national household sample of adults aged 51-61 in 1992, as a data source. STUDY DESIGN: We conducted cross-sectional analyses on the baseline HRS data, and longitudinal analyses using data from eight years of follow-up. We used two-part regression models to estimate the adjusted impact of diabetes on workforce participation, and then estimated the economic impact of diabetes-related losses in productivity. PRINCIPAL FINDINGS: Diabetes is a significant predictor of lost productivity. The incremental lost income due to diabetes by 1992 was 60.0 billion US dollars over an average diabetes duration of 9.7 years. From 1992 to 2000, diabetes was responsible for 4.4 billion US dollars in lost income due to early retirement, 0.5 billion US dollars due to increased sick days, 31.7 billion US dollars due to disability, and 22.0 US dollars billion in lost income due to premature mortality, for a total of 58.6 billion dollars in lost productivity, or 7.3 billion US dollars per year. CONCLUSIONS: In the U.S. population of adults born between 1931 and 1941, diabetes is associated with a profound negative impact on economic productivity. By 1992, an estimated 60 billion US dollars in lost productivity was associated with diabetes; additional annual losses averaged 7.3 billion US dollars over the next eight years, totaling about 120 billion US dollars by the year 2000. Given the rising prevalence of diabetes, these costs are likely to increase substantially unless countered by better public health or medical interventions. PMID- 15533181 TI - Elevated depressive symptoms among caregiving grandparents. AB - OBJECTIVE: To determine whether caregiving grandparents are at an increased risk for depressive symptoms. DATA SOURCE: National sample (n=10,293) of grandparents aged 53-63 years in 1994, and their spouse/partners, who took part in the Health and Retirement Study (HRS). STUDY DESIGN: Grandparents were surveyed in 1994 and resurveyed every two years thereafter, through 2000. Over that period, 977 had a grandchild move in or out of their home. These grandparents served as their own controls to assess the impact of having a grandchild in the home. Data Extraction. Depressive symptoms were measured using an abbreviated form of the Center for Epidemiologic Studies-Depression (CES-D) scale, scored 1-8, with a score > or =4 associated with depression "caseness". PRINCIPAL FINDINGS: At the time of the 1994 interview, 8.2 percent of grandparents had a grandchild in their home. However, there was substantial variation across demographic groups (e.g., 29.4 percent of single nonwhite grandmothers, but only 2.0 percent of single white grandfathers had a grandchild in residence). The impact of having a grandchild in the home varied by grandparent demographic group, with single grandparents and those without coresident adult children experiencing the greatest probability of elevation in depressive symptoms when a grandchild was in residence. For example, single nonwhite grandmothers experienced an 8 percentage point increase in the probability of having a CES-D score > or =4 when a grandchild was in their home, compared to when a grandchild was not in their home, controlling for changes in health care, income, and household composition over time (95 percent CI=0.1 to 15.0 percentage points). CONCLUSIONS: Grandparents have a greater probability of elevated depressive symptoms when a grandchild is in their home, versus when a grandchild is not in their home. Single women of color bear a disproportionate burden of the depression associated with caring for grandchildren. Since an increasing number of grandparents function as a de facto safety net keeping their grandchildren out of formal foster care, identifying strategies to support the health and well-being of caregiving grandparents is an emerging priority. PMID- 15533182 TI - Effective HIV treatment and the employment of HIV(+) adults. AB - OBJECTIVE: To examine whether highly active antiretroviral therapy (HAART) helps HIV-infected patients return to work, remain employed, and maintain hours of work. DATA SOURCE: Longitudinal data from a national probability sample of HIV+ patients older than 18 years old who made at least one visit in the contiguous United States in early 1996. STUDY DESIGN: We consider the effect of HAART on three employment outcomes: (1) returning to work within six months of treatment, conditional on not working pretreatment; (2) remaining employed within six months of treatment, conditional on working pretreatment; (3) hours of work conditional on working at the second follow-up survey. We use a bivariate probit model to jointly model employment and treatment with HAART for the first two outcomes and the two-stage least squares method for hours of work. State policies regarding prescription drug coverage are used as instrumental variables for HAART to account for a key source of potential bias-the more severely ill tend to have the most difficulty working, but are also the most likely to be on HAART. PRINCIPAL FINDINGS: Our results indicate that HAART increases the probability of remaining employed by HIV patients and hours of work for those working within six months of treatment. In the case of remaining employed, the employment effect (an increase from 58 percent to 94 percent in the probability of remaining employed) is statistically significant and the related incremental income is sizable compared to the incremental costs of HAART. Sensitivity analyses demonstrate that the results are robust to different specifications for insurance coverage. CONCLUSIONS: Patients who are working are more likely to remain employed because of treatment with HAART. HAART prescribed to patients in less advanced stages of the infection may lead to the greatest gain in employment. PMID- 15533183 TI - Predicting lawsuits against nursing homes in the United States, 1997-2001. AB - OBJECTIVES: To examine how nursing home characteristics impacted the number of lawsuits filed against the facilities in the United States during 1997-2001. DATA SOURCES/STUDY SETTING: A stratified random sample of 2,378 nursing home in 45 states from 1997-2001. Data were obtained from Westlaw's Adverse Filings: Lawsuits database, the Centers for Medicare and Medicaid Services' (CMS) Online Survey, Certification, and Reporting (OSCAR) database, state complaint surveys, and through primary data. STUDY DESIGN: Negative binomial regression was used to explain total lawsuit variance by year. Explanatory variables included (a) facility characteristics-including staffing, number of beds, multistate system membership, for-profit ownership, (b) quality indicators-including total number and type of quality survey deficiencies, pressure sore development, and (c) market area-state has resident rights statutes, state complaint information. Resident acuity levels and year effects were controlled for. DATA COLLECTION/EXTRACTION METHODS: Nursing homes were identified and linked to Westlaw data that was searched for the number of lawsuits filed against the home, and then linked to OSCAR data and a primary data analysis of multistate chain membership. PRINCIPAL FINDINGS: Staffing levels for certified nursing assistants (CNAs) and registered nurses (RNs) and multistate chain membership were negatively related with higher numbers of lawsuits. More deficiencies on the licensing survey, larger, for-profit nursing homes, and being located in resident rights states were positively related with higher numbers of lawsuits. CONCLUSION: This study suggests that nursing homes that meet long-stay staffing standards and minimum quality indicators, are nonprofit, smaller, and not located in resident rights states will experience fewer lawsuits. PMID- 15533184 TI - An algorithm for the use of Medicare claims data to identify women with incident breast cancer. AB - OBJECTIVE: To develop and validate a clinically informed algorithm that uses solely Medicare claims to identify, with a high positive predictive value, incident breast cancer cases. DATA SOURCE: Population-based Surveillance, Epidemiology, and End Results (SEER) Tumor Registry data linked to Medicare claims, and Medicare claims from a 5 percent random sample of beneficiaries in SEER areas. STUDY DESIGN: An algorithm was developed using claims from 1995 breast cancer patients from the SEER-Medicare database, as well as 1995 claims from Medicare control subjects. The algorithm was validated on claims from breast cancer subjects and controls from 1994. The algorithm development process used both clinical insight and logistic regression methods. DATA EXTRACTION: Training set: Claims from 7,700 SEER-Medicare breast cancer subjects diagnosed in 1995, and 124,884 controls. Validation set: Claims from 7,607 SEER-Medicare breast cancer subjects diagnosed in 1994, and 120,317 controls. PRINCIPAL FINDINGS: A four-step prediction algorithm was developed and validated. It has a positive predictive value of 89 to 93 percent, and a sensitivity of 80 percent for identifying incident breast cancer. The sensitivity is 82-87 percent for stage I or II, and lower for other stages. The sensitivity is 82-83 percent for women who underwent either breast-conserving surgery or mastectomy, and is similar across geographic sites. A cohort identified with this algorithm will have 89-93 percent incident breast cancer cases, 1.5-6 percent cancer-free cases, and 4-5 percent prevalent breast cancer cases. CONCLUSIONS: This algorithm has better performance characteristics than previously proposed algorithms. The ability to examine national patterns of breast cancer care using Medicare claims data would open new avenues for the assessment of quality of care. PMID- 15533185 TI - The effect of area HMO market share on cancer screening. AB - OBJECTIVE: Managed care may have widespread impacts on health care delivery for all patients in the areas where they operate. We examine the relationship between area managed care activity and screening for breast, cervical, and prostate cancer among patients enrolled in more managed care plans and patients who are enrolled in less managed plans. DATA AND METHODS: Data on cancer screening from the 1996 Medical Expenditure Panel Survey (MEPS) were linked to data on health maintenance organization (HMO) and preferred provider organization (PPO) market share and HMO competition at the metropolitan statistical area (MSA) level. Logistic regression analysis was used to examine the relationship between area managed care prevalence and the use of mammography, clinical breast examination, Pap smear, and prostate cancer screening in the past two years, controlling for important covariates. RESULTS: Among all patients, increases in area-level HMO market share are associated with increases in the appropriate use of mammography, clinical breast exam, and Pap smear (OR for high relative to low managed care areas are 1.75, p < .01, for mammography, 1.58, p < .05, for clinical breast exam, and 1.71, p < .01, for Pap smear). In analyses of subgroups, the relationship is significant only for individuals who are enrolled in the nonmanaged plans; there is no relationship for individuals in more managed plans. No relationship is observed between area HMO market share and prostate cancer screening in any analysis. Neither the level of competition between area HMOs nor area PPO market share is associated with screening rates. CONCLUSIONS: Area-level managed care activity can influence preventive care treatment patterns. PMID- 15533186 TI - Care following acute myocardial infarction in the Veterans Administration Medical Centers: a comparison with Medicare. AB - OBJECTIVE: To compare patients treated for acute myocardial infarction (AMI) in a Veterans Health Administration (VHA) facility to similar patients treated under Medicare. DATA SOURCES: Administrative data on 13,129 elderly male veterans hospitalized for AMI in a VHA facility between October 1, 1996, and September 30, 1999, and a matched set of male Medicare beneficiaries with AMI treated in a non VHA facility during the same time period. STUDY DESIGN: We conducted a retrospective cohort study using propensity score methods to identify a matched set of male elderly AMI patients treated either in a VHA facility or in a non-VHA facility under Medicare. We compared the two groups of patients according to characteristics of the admitting hospital, distances traveled for care, the use of invasive procedures, and mortality. We assessed the robustness of our conclusions to biases arising from unmeasured confounders using sensitivity analyses. PRINCIPAL FINDINGS: VHA patients were significantly less likely than Medicare beneficiaries to be admitted to high-volume facilities (for example, 25 percent versus 46 percent in 1999, p<0.001) or facilities with the capability to perform invasive cardiac procedures. Compared to Medicare patients, VHA patients traveled almost twice as far to their admitting hospital. The VHA patients were significantly less likely to undergo coronary angiography or revascularization in the 30 days following their AMI (p<0.001 for all comparisons). Veterans treated in the VHA had significantly higher mortality at one-year in all years studied (for example, 35.2 percent versus 30.6 percent in 1999). The proportion of elderly VHA patients admitted to high-volume facilities increased and 30-day mortality rates decreased between 1997 and 1999. Using sensitivity analyses to assess possible effects of unmeasured confounders, we could explain some but not all of the observed mortality differences. CONCLUSIONS: We observed differences in the way care for AMI patients was structured, in the use of invasive therapies, and in long term mortality between patients treated in VHA hospitals and those treated in non-VHA facilities under Medicare. Future research should focus on explanations for the differences between the two systems and for the reduction in short-term mortality among VHA patients. Further study of these differences both between and within the systems of care may help identify cost effective strategies to improve care in both sectors. PMID- 15533187 TI - Measuring the quality of the VA Health Care System. PMID- 15533188 TI - Understanding employee awareness of health care quality information: how can employers benefit? AB - OBJECTIVE: To analyze the factors associated with employee awareness of employer disseminated quality information on providers. DATA SOURCES: Primary data were collected in 2002 on a stratified, random sample of 1,365 employees in 16 firms that are members of the Buyers Health Care Action Group (BHCAG) located in the Minneapolis-St. Paul region. An employer survey was also conducted to assess how employers communicated the quality information to employees. STUDY DESIGN: In 2001, BHCAG sponsored two programs for reporting provider quality. We specify employee awareness of the quality information to depend on factors that influence the benefits and costs of search. Factors influencing the benefits include age, sex, provider satisfaction, health status, job tenure, and Twin Cities tenure. Factors influencing search costs include employee income, education, and employer communication strategies. We estimate the model using bivariate probit analysis. DATA COLLECTION: Employee data were collected by phone survey. PRINCIPAL FINDINGS: Overall, the level of quality information awareness is low. However, employer communication strategies such as distributing booklets to all employees or making them available on request have a large effect on the probability of quality information awareness. Employee education and utilization of providers' services are also positively related to awareness. CONCLUSIONS: This study is one of the first to investigate employee awareness of provider quality information. Given the direct implications for medical outcomes, one might anticipate higher rates of awareness regarding provider quality, relative to plan quality. However, we do not find empirical evidence to support this assertion. PMID- 15533189 TI - The magnitude and nature of risk selection in employer-sponsored health plans. AB - OBJECTIVE: To determine whether health maintenance organizations (HMOs) attract enrollees who use relatively few medical resources and whether a simple risk adjustment system could mitigate or eliminate the inefficiency associated with risk selection. DATA SOURCES: The first and second rounds of the Community Tracking Study Household Survey (CTSHS), a national panel data set of households in 60 different markets in the United States. STUDY DESIGN: We use regression analysis to examine medical expenditures in the first round of the survey between enrollees who switched plan types (i.e., from a non-HMO plan to an HMO plan, or vice versa) between the first and second rounds of the survey versus enrollees who remained in their original plan. The dependent variable is an enrollee's medical resource use, measured in dollars, and the independent variables include gender, age, self-reported health status, and other demographic variables. DATA COLLECTION METHODS: We restrict our analysis to the 6,235 non-elderly persons who were surveyed in both rounds of the CTSHS, received health insurance from their employer or the employer of a household member in both years of the survey, and were offered a choice of an HMO and a non-HMO plan in both years. PRINCIPAL FINDINGS: We find that people who switched from a non-HMO to an HMO plan used 11 percent fewer medical services in the period prior to switching than people who remained in a non-HMO plan, and that this relatively low use persisted once they enrolled in an HMO. Furthermore, people who switched from an HMO to a non-HMO plan used 18 percent more medical services in the period prior to switching than those who remained in an HMO plan. CONCLUSIONS: HMOs are experiencing favorable risk selection and would most likely continue to do so even if employers adjusted health plan payments based on enrollees' gender and age because the selection is based on enrollee characteristics that are difficult to observe, such as preferences for medical care and health status. PMID- 15533190 TI - Specificity and sensitivity of claims-based algorithms for identifying members of Medicare+Choice health plans that have chronic medical conditions. AB - OBJECTIVE: To examine the effects of varying diagnostic and pharmaceutical criteria on the performance of claims-based algorithms for identifying beneficiaries with hypertension, heart failure, chronic lung disease, arthritis, glaucoma, and diabetes. STUDY SETTING: Secondary 1999-2000 data from two Medicare+Choice health plans. STUDY DESIGN: Retrospective analysis of algorithm specificity and sensitivity. DATA COLLECTION: Physician, facility, and pharmacy claims data were extracted from electronic records for a sample of 3,633 continuously enrolled beneficiaries who responded to an independent survey that included questions about chronic diseases. PRINCIPAL FINDINGS: Compared to an algorithm that required a single medical claim in a one-year period that listed the diagnosis, either requiring that the diagnosis be listed on two separate claims or that the diagnosis to be listed on one claim for a face-to-face encounter with a health care provider significantly increased specificity for the conditions studied by 0.03 to 0.11. Specificity of algorithms was significantly improved by 0.03 to 0.17 when both a medical claim with a diagnosis and a pharmacy claim for a medication commonly used to treat the condition were required. Sensitivity improved significantly by 0.01 to 0.20 when the algorithm relied on a medical claim with a diagnosis or a pharmacy claim, and by 0.05 to 0.17 when two years rather than one year of claims data were analyzed. Algorithms that had specificity more than 0.95 were found for all six conditions. Sensitivity above 0.90 was not achieved all conditions. CONCLUSIONS: Varying claims criteria improved the performance of case-finding algorithms for six chronic conditions. Highly specific, and sometimes sensitive, algorithms for identifying members of health plans with several chronic conditions can be developed using claims data. PMID- 15533191 TI - Identifying and accommodating statistical outliers when setting prospective payment rates for inpatient rehabilitation facilities. AB - OBJECTIVE: To demonstrate how a Bayesian outlier accommodation model identifies and accommodates statistical outlier hospitals when developing facility payment adjustments for Medicare's prospective payment system for inpatient rehabilitation care. DATA SOURCES/STUDY SETTING: Administrative data on costs and facility characteristics of inpatient rehabilitation facilities (IRFs) for calendar years 1998 and 1999. STUDY DESIGN: Compare standard linear regression and the Bayesian outlier accommodation model for developing facility payment adjustors for a prospective payment system. DATA COLLECTION: Variables describing facility average cost per case and facility characteristics were derived from several administrative data sources. PRINCIPAL FINDINGS: Evidence was found of non-normality of regression errors in the data used to develop facility payment adjustments for the inpatient rehabilitation facilities prospective payment system (IRF PPS). The Bayesian outlier accommodation model is shown to be appropriate for these data, but the model is largely consistent with the standard linear regression used in the development of the IRF PPS payment adjustors. CONCLUSIONS: The Bayesian outlier accommodation model is more robust to statistical outlier IRFs than standard linear regression for developing facility payment adjustments. It also allows for easy interpretation of model parameters, making it a viable policy alternative to standard regression in setting payment rates. PMID- 15533192 TI - America's Health Centers: reducing racial and ethnic disparities in perinatal care and birth outcomes. AB - OBJECTIVE: To examine whether community health centers (CHCs) reduce racial/ethnic disparities in perinatal care and birth outcomes, and to identify CHC characteristics associated with better outcomes. BACKGROUND: Despite great national wealth, the U.S. continues to rank poorly relative to other industrialized nations on infant mortality and other birth outcomes, and with wide inequities by race/ethnicity. Disparities in primary care (including perinatal care) may contribute to disparities in birth outcomes, which may be addressed by CHCs that provide safety-net medical services to vulnerable populations. METHODS: Data are from annual Uniform Data System reports submitted to the Bureau of Primary Health Care over six years (1996-2001) by about 700 CHCs each year. RESULTS: Across all years, about 60% of CHC mothers received first trimester prenatal care and more than 70% received postpartum and newborn care. In 2001, Asian mothers were the most likely to receive both postpartum and newborn care (81.7% and 80.3%), followed by Hispanics (75.0% and 76.3%), blacks (70.8% and 69.9%), and whites (70.7% and 66.7%). In 2001, blacks had higher rates of low birth weight (LBW) babies (10.4%), but the disparity in rates for blacks and whites was smaller in CHCs (3.3 percentage points) compared to national disparities for low-socioeconomic status mothers (5.8 percentage points) and the total population (6.2 percentage points). In CHCs, greater perinatal care capacity was associated with higher rates of first-trimester prenatal care, which was associated with a lower LBW rate. CONCLUSION: Racial/ethnic disparities in certain prenatal services and birth outcomes may be lower in CHCs compared to the general population, despite serving higher-risk groups. Within CHCs, increasing first-trimester prenatal care use through perinatal care capacity may lead to further improvement in birth outcomes for the underserved. PMID- 15533193 TI - Hospital choice of rural Medicare beneficiaries: patient, hospital attributes, and the patient-physician relationship. AB - OBJECTIVE: To examine how patient and hospital attributes and the patient physician relationship influence hospital choice of rural Medicare beneficiaries. DATA SOURCES: Medicare Current Beneficiary Survey (MCBS), Health Care Financing Administration (HCFA) Provider of Services (POS) file, American Hospital Association (AHA) Annual Survey, and Medicare Hospital Service Area (HSA) files for 1994 and 1995. STUDY DESIGN: The study sample consisted of 1,702 hospitalizations of rural Medicare beneficiaries. McFadden's conditional logit model was used to analyze hospital choices of rural Medicare beneficiaries. The model included independent variables to control for patients' and hospitals' attributes and the distance to hospital alternatives. PRINCIPAL FINDINGS: The empirical results show strong preferences of aged patients for closer hospitals and those of greater scale and service capacity. Patients with complex acute medical conditions and those with more resources were more likely to bypass their closest rural hospitals. Beneficiaries were more likely to bypass their closest rural hospital if they had no regular physician, had a shorter patient-physician tie, were dissatisfied with the availability of health care, and had a longer travel time to their physician's office. CONCLUSIONS: The significant influences of patients' socioeconomic, health, and functional status, their satisfaction with and access to primary care, and their strong preferences for certain hospital attributes should inform federal program initiatives about the likely impacts of policy changes on hospital bypassing behavior. PMID- 15533195 TI - The placebo response complex. AB - Placebo effects contribute to beneficial therapeutic responses and are common in anxiety and depressive disorders. It is posited that placebo effects are yielded by autonomous feeling-toned complexes capable of re-establishing background self states of well-being. The relationship between the placebo response complex and modern neurobiological models of self is explored. The psychological roots of the placebo response complex in implicit memories of organized attachment between child and early caretakers and in Sandler's conception of the benign superego are examined. The relationships between the negative placebo (nocebo) response complex, Freud's negative therapeutic reaction, and Fordham's defence of the self are explored. Finally, it is suggested that approaches fundamental to the analytic encounter, e.g., mirroring, affectual exchanges, attunement, and containment are likely to optimize the salutary effects of both psychological and somatic therapeutic interventions. PMID- 15533196 TI - Self creation and the limitless void of dissociation: the 'as if' personality. AB - The concept of the 'as if' personality has been used variously in analytic literature without having formed part of a clinically based theoretical development over time. The author discusses the bases of her notion of the 'as if' personality, as observed across a number of patients and supervised patients in intensive, long term analytic treatment. In this composite clinical picture, a grouping of elements that form a particular kind of defence of the self is identified in certain patients with an exceptional capacity for creative engagement in the world, surpassing expectations given their background. The picture includes the presence of physical breakdown and illness, as psychic suffering arising from early narcissistic wounding and from a physical, emotional and/or sexual abusive familial environment, was held for too long in bodily memory but not in mind. A distinction is made between the 'as if' personality, the persona and the false self. The 'as if' personality concerns the action of defensive dissociation deriving from very early experiences of internalizing the presence of an absent object, creating the sense of an internal void at the core of the self. At the same time, the self is capable of acts of self creation through a succession of identifications and internalizations with other sources of environmental nourishment, which substitute for, and are constructed around, the original sense of internal emptiness. Thus are restored, but only up to a point, the resources of the originally diminished self. Until these resources have been used up, the self is often able to excel in activities to an exceptional degree. The countertransference is shown to be the means of both useful but often perilously obtained clinical experience and information, supporting the work along the hazardous analytic journey. PMID- 15533197 TI - The mind beyond our immediate awareness: Freudian, Jungian, and cognitive models of the unconscious. AB - Several branches of cognitive science now focus on the nature of the unconscious. This paper explores some of the findings and models from this research. By introducing formulations based on non-clinical data, the cognitive scientists--in neural linguistics, computational modelling, and neuroscience--may depart from the older psychoanalytic formulations. An understanding of unconscious neural processes is nevertheless emerging showing how synapses are modified by experience and how learning, conscious and unconscious, is due to this important aspect of brain plasticity. Freud and Jung's formulations about the unconscious psyche, representing the main tenets of depth psychology, are also based on a conception of the mind as extending beyond immediate awareness. However, their models are more hypothetical in that their data, almost exclusively, come from treatments of psychotherapy patients and their verbal accounts. So how do these two conceptions of the unconscious match, where do they differ? And how does the neural understanding in the present research support theories and practices of analytic treatments? PMID- 15533198 TI - Boarding school: the trauma of the 'privileged' child. AB - Sending young children to boarding school may be considered a particularly British form of child abuse and social control. The trauma of the rupture with home may be followed by other ordeals such as emotional deprivation and, in extreme cases, physical and sexual abuse. The taboo on expressing emotion, which is common in such institutions, may lead to an encapsulation of the self. Consequently, the needs of the distressed child/self remain active, but unconscious, within the adult. This maybe disguised by an armoured, and very often socially successful, persona. The psychological interplay, between these two facets of the personality, may be detrimental to intimate relationships. In clinical practice the emotional conflict between a desire for intimacy and anticipated exile comes to the fore. Three examples demonstrate how within the transference this may lead to a dependent and erotic atmosphere, which abruptly changes to sever all connection. Changes in the frame, breaks in analysis, and confessions of emotional need are all points at which vigilance is required if such disturbance in analysis is not to end in its abrupt termination. PMID- 15533199 TI - Beyond synchronicity: the worldview of Carl Gustav Jung and Wolfgang Pauli. AB - While exploring the phenomena of synchronicity, Carl Gustav Jung became acquainted with the quantum physicist Wolfgang Pauli and eventually began a collaboration with him. During that collaboration Jung's study of synchronistic phenomena underwent a considerable change; prior to the collaboration, Jung had stressed mainly the phenomenological and empirical features of synchronistic phenomena, while in association with Pauli, he focused his attention upon their ontological, archetypal character. Pauli, on the other hand, became increasingly sensitive to the philosophical aspects concerning the unconscious. Jung and Pauli's common reflections went far beyond psychology and physics, entering into the realm where the two areas meet in the philosophy of nature. In fact, as a consequence of their collaboration, synchronicity was transformed from an empirical concept into a fundamental explanatory-interpretative principle, which together with causality could possibly lead to a more complete worldview. Exploring the problematic character of the synchronicity concept has a heuristic value because it leads to the reconsideration of the philosophical issues that drove Jung and Pauli to clear up the conceptual background of their thoughts. Within the philosophical worldview arising from Jung and Pauli's discussions about synchronicity, there are many symbolic aspects that go against mainstream science and that represent a sort of criticism to some of the commonly held views of present day science. PMID- 15533200 TI - Psychotherapists dream about their patients. PMID- 15533204 TI - Matrix metalloproteinases (MMPs) in oral diseases. AB - Matrix metalloproteinases (MMPs) are a group of enzymes that in concert are responsible for the degradation of most extracellular matrix proteins during organogenesis, growth and normal tissue turnover. The expression and activity of MMPs in adult tissues is normally quite low, but increases significantly in various pathological conditions that may lead into unwanted tissue destruction, such as inflammatory diseases, tumour growth and metastasis. MMPs have a marked role also in tissue destructive oral diseases. The role of collagenases, especially MMP-8, in periodontitis and peri-implantitis is the best-known example of the unwanted tissue destruction related to increased presence and activity of MMPs at the site of disease, but evidence has been brought forward to indicate that MMPs may be involved also in other oral diseases, such as dental caries and oral cancer. This brief review describes some of the history, the current status and the future aspects of the work mainly of our research groups looking at the presence and activity of various MMPs in different oral diseases, as well as some of the MMP-related aspects that may facilitate the development of new means of diagnosis and treatment of oral diseases. PMID- 15533205 TI - Oro-facial lesions and CD4 counts associated with HIV/AIDS in an adult population in Oyo State, Nigeria. AB - OBJECTIVE: The objective of this study was to define the oro-facial lesions associated with Human Immunodeficiency Virus (HIV)/Acquired Immune Deficiency Syndrome (AIDS) in an adult population in Oyo state, Nigeria and to relate these with the level of immune suppression as measured by the CD4 counts. MATERIALS AND METHOD: The study population consisted of 679 consecutive subjects who were seen at the state-owned blood bank. Information on demography, medical and medication histories were obtained. Oro-facial examinations were carried out according to Greenspan et al [Oral Surg Oral Med Oral Pathol (1992)73:142-144]. HIV sero prevalence status was determined for all patients. CD4+ T-lymphocyte count was carried out for those sero-positive for HIV and 31 randomly selected HIV-negative subjects. Data were analyzed using the chi-square test, Fisher's exact test, Student's t-test and odds ratio where appropriate. RESULTS: Eighty-one (11.9%) of the entire study sample were confirmed HIV positive. The prevalence of specific oral lesions by HIV sero-status revealed that pseudomembranous oral candidiasis and angular cheilitis occurred significantly and more frequently in HIV-positive subjects (33.3 and 21% respectively) than those who were HIV negative (4.3 and 1.8% respectively, P < 0.05). The mean CD4 count of the HIV-positive subjects was 452 cells mm(-3), s.d. 137, while it was 602 cells mm(-3), s.d. 251, for the HIV negatives. The difference was statistically significant (P = 0.000). Forty-four (66.7%) subjects with CD4 counts <500 cells mm(-3) had oro-facial lesions whereas among those with CD4 counts >500 cells mm(-3) only 22 (33.3%) had oro-facial lesions (OR = 4.57). CONCLUSION: The type of oro-facial lesions most commonly associated with HIV/AIDS in Oyo state, Nigeria, has been shown to be pseudomembranous oral candidiasis. This was followed by angular cheilitis. These lesions, although found in HIV-negative subjects, were in a lower proportion as compared with HIV-positive subjects. Mean CD4 counts were lower in HIV-positive subjects and this was associated with greater prevalence of oro-facial candidiasis and angular cheilitis. PMID- 15533206 TI - Enfuvirtide: a new class of antiretroviral therapy for HIV infection. AB - Enfuvirtide is the first of a new class of antiretroviral agents recently approved for the treatment of human immunodeficiency virus (HIV)-1 infection. Present available data suggest that enfuvirtide may be a promising agent for the control of HIV infection in patients who have previously received reverse transcriptase inhibitor and protease inhibitor regimens and who are either intolerant to such drugs and/or who have gone into virological failure. Perhaps the greater limitation to the clinical use of enfuvirtide is the cost, limiting its use in the developing world. PMID- 15533207 TI - Oral findings in coeliac disease and Sjogren's syndrome. AB - OBJECTIVE: Both coeliac disease (CD) and Sjogren's syndrome (SS) have an autoimmune background and increased risk of oral mucosal and dental abnormalities. Individuals suffering concomitantly from CD and SS could even be at a higher risk. STUDY DESIGN: Oral mucosal and dental abnormalities were examined in 20 patients with CD + SS (mean age 61 years) and compared with age- and sex-matched controls with either CD or SS. RESULTS: Oral mucosal changes were most common in SS (80%), followed by CD + SS (65%) and CD (40%). Coeliac-type dental enamel defects were found in 89% in CD + SS and in 88% in CD compared with only 25% in SS (P < 0.001). The median number of teeth was six in the CD + SS, 24 in the CD and 22 in the SS group. The DMF index was higher (P < 0.005) in the CD + SS than in the CD group. CD + SS was characterized by higher salivary flow rate (P < 0.001) and lower inflammatory focus score in the salivary glands (P < 0.01) than SS. CONCLUSIONS: The co-occurrence of CD and SS should be recognized because of its effects on dental and oral mucosal health. A lower salivary gland inflammatory focus score and higher salivary flow rate in CD + SS than in SS suggests that a gluten-free diet treatment may alleviate autoimmune inflammation. PMID- 15533208 TI - The association of tobacco and other factors with recurrent aphthous stomatitis in an US adult population. AB - OBJECTIVE: To determine point and annual prevalence of recurrent aphthous stomatitis (RAS). SETTING: Reported prevalence of RAS in textbooks and much of the literature varies according to study location, patient selection and whether point prevalence (presence of lesions at examination) or period prevalence (history of lesions during a specified period) is reported. Many studies are based on non-probability samples and this may contribute to significant variation in reported prevalence and factors presumed to be associated with RAS. METHODS: We analyzed data from the Third National Health and Nutrition Examination Survey, 1988-1994, a large United States probability sample, for RAS and covariates suggested in the literature using bivariate and multivariate logistic regression. RESULTS: Oral mucosal examinations were performed on 17 235 adults 17 years and older. Of these, 146 (0.89%) had at least one clinically apparent aphthous lesion. For annual (reported) prevalence, Whites (20.87%) and Mexican-Americans (12.88%) had several fold higher prevalence of RAS than Blacks (4.96%). Adults younger than 40 years of age had almost twice the prevalence (22.54%) of those older than 40 years (13.42%). CONCLUSION: Annual prevalence was significantly higher in whites and Mexican-Americans (compared with blacks), individuals 17-39 years of age, cigarette non-smokers, and those with recurrent herpes labialis history; while it was lower in males. Point prevalence was significantly higher in whites, Mexican-American, individuals 17-39 years of age, cigarette non smokers, and males. PMID- 15533209 TI - The prevalence of oral leukoplakia in 138 patients with oral squamous cell carcinoma. AB - OBJECTIVES: To determine the relationship between oral leukoplakia (OL) and oral squamous cell carcinoma (OSCC), and to evaluate possible differences between those carcinomas with and without associated leukoplakia. MATERIAL AND METHODS: A total of 138 patients were studied at the Stomatology Service of the University General Hospital, Valencia, Spain. These patients were divided into two groups: group 1, patients with oral cancer and leukoplakia, and group 2, patients with OSCC but with no associated premalignant lesions. The relationship between this precancerous lesion and the OSCC was evaluated, as well as the possible clinical and histological differences between the tumours of the two groups. RESULTS: Leukoplakia was detected in 27 (19.56%) patients with OSCC. No differences were found between the two groups regarding age and tumour location. However, statistically significant differences were observed with respect to the form, tumour stage and the presence of adenopathies in the cancers with and without leukoplakia; in that the tumours associated with leukoplakia were diagnosed as being at a more initial stage. CONCLUSIONS: Those patients with OL associated with oral cancer presented with tumours at a less advanced stage than those where no associated leukoplakia existed. PMID- 15533210 TI - T102C polymorphism of the 5-HT2A receptor gene may be associated with temporomandibular dysfunction. AB - OBJECTIVE: To assess whether a relationship existed between the T102C polymorphism of 5-HT2A receptor gene and temporomandibular dysfunction. METHODS: Sixty-three patients with temporomandibular dysfunction, and 54 healthy volunteer controls were included in the study. Molecular analysis of the T102C polymorphism of the 5-HT2A receptor gene was performed using PCR technique. RESULTS: The C/C genotype was over represented in the patients whereas T/T genotype was over represented in the controls (P < 0.05). The genotype distribution of the patients who had temporomandibular dysfunction was not different than those who did not have temporomandibular dysfunction (P > 0.05). CONCLUSION: The T102C polymorphism may be involved in the etiology of temporomandibular dysfunction. The overrepresentation of the C/C variant of 5-HT2A receptor gene in temporomandibular dysfunction suggests a possible role of the serotonergic system in this disease, particularly at the receptor level. PMID- 15533211 TI - Linkage analysis between BCL3 and nearby genes on 19q13.2 and non-syndromic cleft lip with or without cleft palate in multigenerational Japanese families. AB - OBJECTIVE: To investigate the linkage between candidate genes on chromosome 19 and cleft lip with or without cleft palate in Japanese using a parametric method. MATERIALS AND METHODS: After informed consent was obtained, blood samples were drawn from 90 individuals in 14 families, 30 of whom were affected, and genomic DNAs were extracted. PCR-amplified products using four microsatellite markers, D19S178, BCL3, APOC2[007/008] and APOC2[AC1/AC2] located in 19q13.2, were separated by 8% polyacrylamide gel electrophoresis. Linkage analysis was carried out using the MLINK and LINKMAP programs, and logarithm of odds (LOD) scores were calculated for each family. RESULTS: Before undertaking linkage analysis, we analyzed 74 healthy Japanese subjects and found racial differences in that the observed number of alleles and their heterozygosity were lower in Japanese than in Caucasians, and that both populations tended to show a different allele distribution. In 14 families, two-point maximum LOD score (Zmax) for BCL3 was 0.341 and multi-point Zmax was less than -2 excluding linkage. But in 9 families with left and bilateral CL/P, two-point Zmax for APOC2[AC1/AC2] was 1.701 and multi-point Zmax at APOC2 locus was 1.909. CONCLUSION: The LOD score was relatively high but provided no evidence of linkage for CL/P to BCL3 and nearby genes in Japanese subjects. PMID- 15533212 TI - Regulation of interleukin-6 expression by arecoline in human buccal mucosal fibroblasts is related to intracellular glutathione levels. AB - OBJECTIVES: Cytokines play an important role in regulating fibroblast function and is likely to play a key role in regulating the initiation and progression of scarring in any fibrotic disease. Interleukin-6 (IL-6) has been implicated in the development of a variety of fibrotic diseases. The aim of this study was to compare IL-6 expression in fibroblasts cultured from normal human buccal mucosa and oral submucous fibrosis (OSF) specimens and further explore the potential mechanism that may lead to induce IL-6 expression. METHODS: mRNA level of IL-6 in fibroblasts from OSF was compared with normal buccal mucosa. The effects of arecoline, the major areca nut alkaloid, on IL-6 expression in normal human buccal mucosa fibroblasts (BMFs) were measured in vitro. mRNA was quantified with AlphaImager 2000. To determine whether glutathione (GSH) levels were important in the induction of IL-6 by arecoline, we pretreated cells with 2-oxothiazolidine-4 carboxylic acid (OTZ) to boost GSH levels or with buthionine sulfoximine (BSO) to deplete GSH. RESULTS: Fibroblasts derived from OSF exhibited higher IL-6 gene expression than BMF in mRNA levels (P < 0.05). The exposure of quiescent BMF to arecoline resulted in the elevation of IL-6 mRNA expression in a dose-dependent manner (P < 0.05). IL-6 gene regulated by arecoline correlated with intracellular GSH levels in BMF. Arecoline at a concentration of 129 muM induced about 2.7-fold IL-6 mRNA levels over the 6-h incubation period. However, BSO enhanced the IL-6 mRNA levels by 3.9-fold (P < 0.05). In addition, OTZ was found to marginally reduce the arecoline-induced IL-6 expression by about 1.7-fold (P < 0.05). CONCLUSIONS: Taken together, these results suggest that IL-6 expression is significantly upregulated in OSF fibroblasts in areca quid chewers and arecoline may be responsible for the enhanced IL-6 expression. In addition, the regulation of IL-6 expression induced by arecoline is critically dependent on the intracellular GSH concentrations. PMID- 15533213 TI - Activities of daily living and chewing ability in an 80-year-old population. AB - OBJECTIVES: The aim of this study was to determine whether chewing ability or number of intact teeth was related to performance of activities of daily living in 80-year olds. DESIGN: This was a cross-sectional survey including dental examinations, chewing self-assessment, and everyday function. SUBJECTS AND METHODS: A total of 823 people (309 males, 514 females) who were 80 years of age and resident in the Fukuoka Prefecture, Japan participated in this study. RESULTS: The mean number of intact teeth was 7.5 +/- 8.7; of the 15 food items offered, the mean number of foods chewed was 11.2 +/- 3.8. Function was independent in 713 subjects, and dependent in 93. After adjustment for gender, we found the number of foods chewed being linked to independent function by the multiple regression analysis (beta = 0.223, P < 0.0001); the number of intact teeth was only weakly related (beta = 0.077, P = 0.03). By logistic regression analysis, functional dependency was 7.5 times more prevalent in individuals capable of chewing four foods or fewer, and 3.3 times more prevalent in those chewing five to nine, than in those chewing all 15 items. No significant functional difference was related to the number of intact teeth. CONCLUSION: Chewing ability may be independently related with functional status in 80-year olds. PMID- 15533214 TI - Radiographic examination of the mandibular (glenoid) fossa in ancient and modern man. AB - OBJECTIVE: To compare the morphology of the mandibular (glenoid) fossa between ancient and modern Japanese. METHODS: There were 221 specimens from the ancient period and 206 specimens from the modern period. Radiographs of the mandibular fossa were obtained using a modified Schuller technique and were studied. The length, depth, and area of the mandibular fossa and angulation of the frontal slope of the mandibular fossa on the radiographs were automatically computed. In addition, morphological changes of the mandibular fossa in the ancient or modern periods on radiographs were evaluated in order to assess tendency of temporomandibular joint disorder (TMD). RESULT: The ratio of mandibular fossa having morphological alternation in the modern period was greater than that in the Yayoi. Differences in the length and area between the right and left sides became progressively larger from the Yayoi to modern periods. The angulation of the anterior slope of the mandibular fossa in the modern period was smaller than that in other periods. CONCLUSION: The morphology of the mandibular fossa was shown to have changed and asymmetry has increased from the Yayoi to modern periods. It can be speculated that this change in morphology of the mandibular fossa in the modern period is the result of an increase of TMD. PMID- 15533215 TI - Comparative morphological analysis of the root developmental groove with the palato-gingival groove. AB - The palato-gingival groove is an anomaly of shape that modifies dental tissues organization while the developmental root groove is described within normal root anatomy. The morphology of dental tissues in relation to the presence of the developmental root groove has not been properly described. This study analyzed microscopically the morphology of dental tissues related to the root developmental groove comparing it with that presented on teeth affected by palato gingival groove. Many similarities were observed such as the increased cementum thickness, decreased dentin thickness, pulp compartment surface alteration, irregularity of the dentin-cementum junction and of the cementum surface. These results suggest a common determining factor to this structure organization pattern. It is possible that the palato-gingival groove could be the result of an alteration of genetic mechanisms, rather than a dental germ folding, determined by privation of space, as previously hypothesized. PMID- 15533216 TI - MMP-2 activation by Actinobacillus actinomycetemcomitans supernatant in human PDL cells was corresponded with reduction of TIMP-2. AB - OBJECTIVE: Matrix metalloproteinase 2 (MMP-2) has been implicated to play a role in pathogenesis of periodontal disease. We recently reported that Porphyromonas gingivalis supernatant could activate MMP-2 in human periodontal ligament (HPDL) cells. In this study, activation of MMP-2 by Actinobacillus actinomycetemcomitans supernatant and the mechanism was investigated. METHODS: HPDL cells were treated with either A. actinomycetemcomitans or P. gingivalis supernatant for 48 h. To verify the mechanism, pretreated inhibitors were used. Gelatin zymography, RT-PCR and Western blot analysis were used to detect the activation of MMP-2, expression of MT1-MMP and TIMP-2 mRNA and the proteins, respectively. RESULTS: The supernatant from A. actinomycetemcomitans could activate MMP-2 in HPDL cells similar to that from P. gingivalis but by a different mechanism. Activation by A. actinomycetemcomitans supernatant was correlated with a reduction of TIMP-2 secretion without any alteration of MT1-MMP, while activation by P. gingivalis increased MT1-MMP but no change of TIMP-2 was found. CONCLUSION: The supernatant from A. actinomycetemcomitans and P. gingivalis could induce the activation of MMP-2 possibly through the imbalance of MT1-MMP and TIMP-2 in HPDL cells but by different mechanisms. The imbalance of MT1-MMP and TIMP-2 may be another factor that is involved in the severity of periodontal disease. PMID- 15533217 TI - Molecular evaluation of residual endodontic microorganisms after instrumentation, irrigation and medication with either calcium hydroxide or Septomixine. AB - BACKGROUND AND OBJECTIVE: The correct choice of antimicrobial agents as inter appointment medicaments is as important as the instrumentation and irrigation to remove pathogens from infected root canals. Calcium hydroxide [Ca(OH)2] and framycetin sulfate (Septomixine) are common endodontic medicaments. Therefore, we evaluated the efficacy of either calcium hydroxide or Septomixine in eliminating residual intra-canal bacteria, particularly Actinomyces spp., during inter appointment interval in endodontic therapy using molecular methods. METHODS: A total of 31 single-rooted teeth with primary root canal infections were studied immediately after opening the canals and subsequently after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 25) or Septomixine (n = 6). Whole bacterial genomic DNA was isolated directly from samples and PCR with universal primers performed to detect total intra-canal bacteria. The variable regions of 16S rDNA of bacteria were amplified and labeled with digoxigenin for further hybridization to detect Actinomyces spp. A total of seven oligonucleotide probes specific for A. bovis, A. gerencseriae, A. israelii, A. meyeri, catalase-negative A. naeslundii (genospecies 1 and 2), catalase positive A. naeslundii genospecies 2 and A. odontolyticus were used to detect Actinomyces spp. in 22 of 31 medicated root canals [Ca(OH)2: n = 17; Septomixine: n = 5]. RESULTS: The PCR results showed that 25 of 31 examined canals were positively detected with residual microorganisms after instrumentation, irrigation with sterile saline and 1-week medication with either Ca(OH)2 (n = 20) or Septomixine (n = 5). Thus, only six canals [Ca(OH)2: n = 5, Septomixine: n = 1] were aseptic after treatment. Hybridization results showed higher detection frequency of both A. odontolyticus and A. gerencseriae after treatment. Significant correlation was found between exposed pulp before treatment and positive detection of Actinomyces spp., particularly A. odontolyticus on the second visit (P < 0.05). CONCLUSION: The conventional, 1-week medication of either Ca(OH)2 or Septomixine in endodontic therapy may not effectively inhibit residual bacterial growth in all root canals during inter-appointment intervals. Further investigations using, for instance quantitative real-time PCR analyses, are required to substantiate the present findings. PMID- 15533218 TI - Liposarcoma circumscriptum (lipoma-like) of the tongue: report of a case. AB - Liposarcoma, first described by Virchow in 1857, is the second most frequent sarcoma of soft tissues, although it is rare both in the head and neck and the oral cavity. Intra-orally, liposarcoma has been reported in the jawbones (particularly the maxilla) and the soft tissues -- mainly the cheek and floor of mouth, but it is rare in the tongue. A case of well-differentiated, superficial liposarcoma circumscriptum of the tongue is reported. The authors underline the difficulties in the clinical and histopathological diagnosis, as this tumour may be confused with lipoma. In view of the indolent behaviour of this tumour type, local recurrence or metastasis rarely occur; consequently, conservative surgical therapy is advised without adjuvant chemo-radio therapy. PMID- 15533219 TI - Adverse oral reactions associated with the COX-2 inhibitor rofecoxib. AB - Rofecoxib is an inhibitor of the enzyme cyclo-oxygenase 2 (COX-2) used as an analgesic and anti-inflammatory agent especially in rheumatoid arthritis and osteoarthritis. The adverse effects are generally less than with other anti inflammatory drugs (NSAIDs), and no oral adverse reactions have been reported in the literature. We present three cases of oral lesions caused by rofecoxib. Extensive erosions appeared, which in all cases resolved on withdrawal of the drug. PMID- 15533221 TI - Myxoid neurothekeoma of the oral mucosa: an unusual benign tumor. PMID- 15533220 TI - Poland-Moebius syndrome: a case with oral anomalies. AB - The combination of Moebius and Poland anomalies is rarely described in the literature. While some authors believe this association is an independent syndrome, others think that Poland, Moebius and Poland-Moebius syndromes are variations of the same condition. We report a case of Poland-Moebius syndrome in a 6-year-old girl who presented with bilateral convergent strabismus, the inability to abduct her eyes beyond the midline and brachydactyly of her right hand. Oral manifestations included incompetent lips and an abnormal tongue. Other facial features included hypoplasia of her mandible and her left ear at a slightly lower level than her right ear. Panoramic and lateral skull radiographs confirmed the absence of certain teeth and the hypoplastic mandible. The diagnosis of Poland-Moebius syndrome was made on the basis of cranial nerve involvement and oro-facial manifestations. PMID- 15533223 TI - Higher frequency of p53 gene mutations in diffuse large B-cell lymphoma with MALT component. AB - p53 gene mutation is not a frequent event in the tumorigenesis of lymphomas and the expression of p53 protein is independent of p53 gene mutations. The present study aimed to investigate mutations in the p53 gene in a series of extranodal B cell lymphomas, and its association with p53 protein expression. A total of 52 cases were graded histologically into Grade 1, Grade 2 and Grade 3 tumors and p53 protein expression was detected using immunohistochemistry. Mutations in the p53 gene were analyzed using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and mobility shifts were confirmed by direct sequencing. The tumors comprised 26 (50%) Grade 1, 9 (17%) Grade 2 and 15 (29%) Grade 3. A high proportion of Grade 2 (25%) tumors expressed p53 protein (P = 0.051) and carried p53 gene mutation (33%) (P = 0.218). However, p53 protein expression was not associated with p53 gene mutations (P = 0.057). Transversion mutations (88%) were more frequently detected than transition mutations (12%). The present study revealed that p53 gene mutations and p53 protein expression occurred in higher frequencies in Grade 2 tumors, which may be of pathogenetic importance. The high frequency of transversion mutations may reflect the influence of an etiological agent in the tumorigenesis of mucosa-associated lymphoid tissue (MALT lymphoma). PMID- 15533224 TI - Hepatic cytotoxic T-cell infiltrates in patients with peripheral T-cell proliferative diseases/lymphomas: clinicopathological and molecular analysis. AB - Seventy patients with various types of peripheral T-cell proliferative disease/lymphoma who manifested with prolonged fever, weight loss, anemia, lymphadenopathy, hepatosplenomegaly and elevated serum levels of alkaline phosphatase and/or lactate dehydrogenase were evaluated. Histopathological examination of the livers revealed T-cell infiltration into the hepatic sinusoids and portal tracts. The morphology of the infiltrated T cells varied from mature small lymphocytes to malignant lymphoid cells. The liver pathology was classified into four groups on the basis of cellular atypia. Group A and group B showed mature lymphoid cell infiltration; however, only group B had multiple large areas of hepatocellular necrosis. Group C showed atypical lymphoid cell infiltration and in group D malignant lymphoid cell infiltrates were demonstrated. The majority of the antigenic phenotypes of these T-cell infiltrates were CD3+, CD4-, CD8+, CD20-, CD45RO+, CD56-, CD57-, TIA-1+ and betaF1-. Epstein-Barr virus RNA in the nuclei of the infiltrated T cells was recorded in 38.6% of the patients and was more common in groups C and D. Patients in groups B, C and D had a very poor prognosis, median survival was only 1 month, whereas median survival in group A patients was 36 months. Chemotherapy was not effective in improving survival. Monoclonal band/s of T-cell receptors (TCR) beta and/or gamma gene rearrangements were detected in 88.6% of patients, and DNA-sequence analysis showed high identity to the human TCR germline gene. PMID- 15533225 TI - Clinicopathological features of eight Korean cases of primary hepatic lymphoma. AB - Primary hepatic lymphoma is very rare, accounting for less than 0.4% of extranodal lymphomas. Furthermore, hepatic lymphoma, either primary or metastatic, is infrequently confirmed histopathologically in needle biopsy specimens. The aim of the current study is to assess the clinicopathological characteristics of primary hepatic lymphomas in Korea, which is an endemic area of chronic B viral hepatitis. In total, 17 cases with liver needle biopsy specimens with involvement of malignant lymphoma, from whom eight cases met the criteria for primary hepatic lymphoma, were selected. The clinicopathological features were reviewed. Five of eight (62.5%) cases were T cell lymphoma, including three cases (37.5%) of hepatosplenic T cell lymphoma. Three cases (37.5%) were diffuse large B cell lymphomas. Seven patients had follow-up data from 25 days to 50 months that was available for evaluation. The partial remission was present in two of seven patients (28.6%) and five patients (71%) died of disease 25 days to 7 months after the diagnosis. The data indicate that the relatively high incidence of T-cell type in Korean cases of primary hepatic lymphoma may be related to its aggressive behavior and poor prognosis despite combination chemotherapy. PMID- 15533226 TI - Glomerular crescents are responsible for chronic graft dysfunction in post transplant IgA nephropathy. AB - Information is limited regarding the histological features related to chronic graft dysfunction and failure in patients with IgA nephropathy developing after renal transplantation. The prevalence and significance of glomerular crescents in post-transplant IgAN including recurrent, de novo and transmitted forms (TxIgAN), were studied. Renal morphology was evaluated in 71 patients of TxIgAN, obtained at more than 6 months post-transplant, and compared with regard to the presence (C-TxIgAN) or absence (N-TxIgAN) of glomerular crescents. Crescents were demonstrated in 12 samples of 10 patients (14.1%). The percentages of crescents were from 4.8% to 83.3% (median, 28.6%) in each sample. Ten samples of C-TxIgAN had cellular to fibrocellular crescents, and four of these were associated with diffuse mesangial proliferation. Serum creatinine levels and the frequency of nephrotic range proteinuria at the time of biopsy and the degree of interstitial inflammation were significantly different in the two groups. Graft survival after allograft biopsies was significantly lower in C-TxIgAN (P = 0.0017). Chronic rejection was a major cause of graft loss in N-TxIgAN (31.8%), whereas TxIgAN was the major cause in C-TxIgAN (66.7%). In conclusion, the current study suggests that glomerular crescents are not rare and that they are responsible for chronic graft dysfunction in TxIgAN patients. PMID- 15533227 TI - Hypoxia-inducible factor-2alpha is involved in enhanced apoptosis in the placenta from pregnancies with fetal growth restriction. AB - The aim of the present study is to investigate whether hypoxia-inducible factors (HIF-1alpha, HIF-2alpha, HIF-1beta) are involved in enhanced apoptosis in the human placenta from pregnancies with fetal growth restriction (FGR). Placental samples were obtained from women with normal term pregnancy (n = 18) or from pregnancy with FGR (n = 12). Placenta apoptosis was assessed by the terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling (TUNEL) staining. The expressions of HIF-1alpha, HIF-2alpha, and HIF-1beta were examined by immunohistochemical analysis. Enhanced apoptosis was observed in the placenta from pregnancies with FGR compared with normal term placenta. The apoptosis index in FGR group (1.45 +/- 1.26%) was significantly higher than that in the normal control group (0.18 +/- 0.16%; P < 0.01). There were no significant differences in the intensity of the staining for HIF-1alpha and HIF-1beta expressions between two groups, while HIF-2alpha was overexpressed in the placenta from pregnancies with FGR group (P < 0.05). The upregulation of HIF-2alpha protein expression in the placenta from pregnancies with FGR may, at least in part, be involved in the increased placental apoptosis. PMID- 15533228 TI - Multiple granulomatous inflammation in the minor salivary glands: a proposed new entity, allergic granulomatous sialadenitis. AB - We report a patient who presented with multiple small submucosal nodules with granulomatous inflammation in the minor salivary glands of the oral cavity. A 43 year-old woman presented with a 1-week history of multiple small submucosal nodules in her oral cavity after having taken medicine for abdominal pain. The patient did not have a history of fever, rectal bleeding, skin lesions or arthritis, but did have a history of drug allergy and bronchial asthma. Histopathological examination of the submucosal nodules showed sialadenitis with marked infiltration of lymphocytes, eosinophilic cells, macrophages and Langhans type or foreign-body-type multinucleate giant cells. The macrophages tended to be aggregated and appeared to have caused immature granuloma formation without caseous necrosis. Degranulated eosinophilic cells were numerous. Sarcoidosis, Crohn's disease, tuberculosis and atypical mycobacterial infection were not identified by medical examination. Three weeks after discontinuing the medication the patient was seen again at a follow-up visit. Multiple submucosal small nodules and other symptoms were not evident at that time. This case report may represent a new entity of salivary gland disease that we tentatively refer to as 'allergic granulomatous sialadenitis'. PMID- 15533229 TI - Extremely well-differentiated adenocarcinoma of the gastric cardia: a unique case with columnar cells and laminated stones. AB - Extremely well-differentiated adenocarcinoma (EWDA) is an unusual gastric cancer that is histologically too bland to be diagnosed as malignant neoplasm, particularly using biopsy. EWDA may be a gastric counterpart of 'adenoma malignum' or minimal deviation adenocarcinoma (MDA) in the uterine cervix; however, the clinicopathological features of EWDA remain less apparent than those of MDA. A 60-year-old male was complaining of dysphagia. He had been made aware of a small submucosal tumor in the cardia 2 years before the onset of this symptom. Endoscopic ultrasonographic examination revealed a large cardiac tumor consisting of thickened layers, as observed in Borrmann type IV. Three mucosal biopsies suggested only benign changes including adenoma and hyperplastic polyps. At the fourth biopsy, cytologically bland columnar cells were located in the submucosa along with stromal fibrosis and laminated stones. The possibility that non-neoplastic aberrant pancreas with lithiasis formed the tumor was denied at laparotomy by a frozen section that revealed benign-looking glands invading the diaphragm. Immunohistochemically the cancer glands were positive for CA19-9 and human gastric mucin, but not for p53 or MUC2. To our knowledge, this is a previously unknown combination of EWDA and psammomatous calcification in the stomach. PMID- 15533230 TI - Hepatocellular carcinoma with an unusual neuroendocrine component. AB - We report a rare case of hepatocellular carcinoma (HCC) with an unusual neuroendocrine component. During a follow-up study for chronic hepatitis C in a 71-year-old man, a nodular lesion showed rapid growth from 1 cm to 4 cm in diameter within 3 months. Histologically, the tumor was consistent with moderately differentiated HCC, but was intermingled with nests of small round cells with scarce cytoplasm, which resembled those found in small cell carcinoma. This population formed small solid nests among the trabecular structures. Immunohistochemically the small round cell component of the tumor was strongly positive for neuron-specific enolase (NSE), chromogranin A and synaptophysin, but hepatocyte paraffin-1 (HP-1) and alpha-fetoprotein (AFP) were negative. In contrast, HP-1 and AFP were positive, and NSE, chromogranin A and synaptophysin were negative in moderately differentiated HCC tissues. Electron microscopy revealed many intracytoplasmic neurosecretory granules in the small round cells. The labeling indexes of p53 and Ki-67 were significantly higher in the small round cell component than in the moderately differentiated HCC component. Overall, we conclude that this nodule was HCC with neuroendocrine differentiation. PMID- 15533231 TI - Sarcomatoid carcinoma with components of small cell carcinoma and undifferentiated carcinoma of the gallbladder. AB - We report a case of sarcomatoid carcinoma with components of small cell carcinoma and undifferentiated carcinoma of the gallbladder. An 84-year-old woman was admitted to our university hospital with right upper abdominal pain and back pain. Clinical diagnosis of a gallbladder tumor was made based on the findings of abdominal ultrasonography, computed tomography and endoscopic retrograde cholangiopancreatography, and a cholecystectomy was carried out. On gross examination a pedunculated polypoid tumor protruded into the lumen of the gallbladder. Histologically the tumor was composed of carcinomatous and sarcomatous components; the carcinomatous component consisted mainly of small cell carcinoma and undifferentiated carcinoma. In general, the carcinomatous component of sarcomatoid carcinoma of the gallbladder consists of adenocarcinoma, and there have only been two previously reported cases in which the carcinomatous component consisted of small cell carcinoma or undifferentiated carcinoma. Because the patient's prognosis may be influenced by the peculiar carcinomatous component in such cases, it is important to accumulate case reports that clarify their clinicopathological features. PMID- 15533232 TI - Balloon cell nevus of the soft palate: an immunohistochemical and ultrastructural study. AB - An ultrastructural analysis of oral balloon cell nevus of intramucosal type complemented with an immunohistochemical study was performed for the first time. The lesion was composed of large balloon cells with an admixture of small nevus cells and melanophages at the periphery. Balloon cells showed cytoplasmic accumulation of vacuoles of varying sizes and the presence of microgranular and vacuolated melanosomes were found. Residual cytoplasm contained no identifiable organelles. A spectrum of transitional forms between balloon cells and conventional nevus cells with microvacuoles was readily observed. Both cells exhibited intense immunoreactivity to multiple melanocytic markers. Ballooning phenomenon was not evident in melanophages containing a large amount of melanosome complex. It can be inferred, from the present and previous observations, that progressive vacuolization of melanosomes in nevomelanocytes may be responsible for the formation of peculiar ballooning appearance, suggesting an aberrant melanogenesis. PMID- 15533233 TI - Sebaceous gland hyperplasia in an intraoral fibrous polyp. PMID- 15533235 TI - Melanocortin 1 receptor variants, pigmentation, and skin cancer susceptibility. AB - The melanocortin 1 receptor is a key regulator of variation in normal human pigmentation. Genetic variants of this receptor cause red hair and fair skin, and several case-control studies have demonstrated that these genetic variants increase the risk of skin cancer development in humans. The mechanism whereby the risks of skin cancer are increased is not entirely clear, and may be because of a combination of effects on pigmentation and non-pigmentary pathways. PMID- 15533236 TI - Iatrogenic skin cancer: induction by psoralen/ultraviolet A and immunosuppression of organ transplant recipients. AB - Photochemotherapy (psoralen/UVA (PUVA)) is an efficient therapeutic tool for a wide range of skin diseases. Concern, however, exists regarding the long-term carcinogenic effects of this treatment modality and, as a consequence, is being used less frequently. PUVA remains an important treatment in our therapeutic armamentarium but must be used with caution in those patients with risk factors and cumulative dose exposure must be limited. PUVA-induced cancers show features in common with skin cancers induced by immunosuppressed organ transplant recipients. Tumours in the latter group of individuals are, however, much more aggressive and difficult to manage. PMID- 15533237 TI - Influence of oral antioxidants on ultraviolet radiation-induced skin damage in humans. AB - Ultraviolet radiation (UVR) causes a range of acute and chronic adverse cutaneous effects, in addition to some beneficial effects. In present times, the skin is generally exposed to higher levels of UVR, such that inherent defence mechanisms become overwhelmed. Complications, notably skin malignancies, show a serious rise in incidence. Since many effects of UVR are mediated through generation of reactive oxygen species, antioxidant supplementation provides a strategy to combat their excess generation, and hence reduce the clinical consequences. Human supplementation studies examining the potential of a range of oral agents to protect against UVR-induced skin effects show mixed results; further studies should examine whether certain subgroups of the population may show augmented benefit. PMID- 15533238 TI - Investigation of photosensitive disorders. AB - Investigation of photodermatosis is based primarily on the history and clinical findings, histological, immunological and biochemical findings are variably helpful depending on the clinical picture. Formal testing for photosensitivity may be the only definitive test proving photosensitivity on occasion. This article delineates the important features which allow classification of patients which then enables appropriate treatment. PMID- 15533239 TI - Chronic actinic dermatitis. PMID- 15533240 TI - Psoralen ultraviolet A therapy in 2004. PMID- 15533241 TI - Isoform-specific expression of the Coxsackie and adenovirus receptor (CAR) in neuromuscular junction and cardiac intercalated discs. AB - BACKGROUND: The Coxsackie and adenovirus receptor (CAR) has a restricted expression pattern in the adult. In skeletal muscle, although CAR is expressed in immature fibers, its transcript levels are barely detectable in mature muscle. This is in contrast to the robust expression observed in the heart. However, both heart and skeletal muscle are susceptible to infection with the Coxsackie B virus which utilizes primarily CAR for cellular internalization. The specific point of viral entry in skeletal and heart muscle remains unknown. RESULTS: Using antibodies directed against the extracellular and the cytoplasmic domains of CAR, we show CAR in normal human and mouse skeletal muscle to be a novel component of the neuromuscular junction. In cardiac muscle, CAR immunoreactivity is observed at the level of intercalated discs. We demonstrate a single isoform of CAR to be expressed exclusively at the human neuromuscular junction whereas both predominant CAR isoforms are expressed at the intercalated discs of non-diseased human heart. CONCLUSION: The localization of CAR to these important junctional complexes suggests that CAR may play both a structural and a regulatory role in skeletal and cardiac muscle, and that these complexes may serve as a point of entry for Coxsackie B virus. PMID- 15533242 TI - A survey of individual preference for colorectal cancer screening technique. AB - BACKGROUND: Due to the low participation in colorectal cancer screening, public preference for colorectal cancer screening modality was determined. METHODS: A cross-sectional survey was performed of healthy ambulatory adults in a pediatrics primary care office and neighboring church. Overall preference was ranked for each of four colorectal cancer screening modalities: Faecal Occult Blood, Fiberoptic Sigmoidoscopy, Barium Enema and Colonoscopy. Four additional domains of preference also were ranked: suspected discomfort, embarrassment, inconvenience and danger of each exam. RESULTS: 80 surveys were analyzed, 57 of which were received from participants who had experienced none of the screening tests. Fecal Occult Blood Testing is significantly preferred over each other screening modality in overall preference and every domain of preference, among all subjects and those who had experienced none of the tests. CONCLUSIONS: Efforts to increase public participation in colorectal cancer screening may be more effective if undertaken in the context of public perceptions of screening choices. PMID- 15533243 TI - Mosquitoes and transmission of malaria parasites - not just vectors. AB - The regional malaria epidemics of the early 1900s provided the basis for much of our current understanding of malaria epidemiology. Colonel Gill, an eminent malariologist of that time, suggested that the explosive nature of the regional epidemics was due to a sudden increased infectiousness of the adult population. His pertinent observations underlying this suggestion have, however, gone unheeded. Here, the literature on Plasmodium seasonal behaviour is reviewed and three historical data sets, concerning seasonal transmission of Plasmodium falciparum, are examined. It is proposed that the dramatic seasonal increase in the density of uninfected mosquito bites results in an increased infectiousness of the human reservoir of infection and, therefore, plays a key role in "kick starting" malaria parasite transmission. PMID- 15533244 TI - Clinical education of ethicists: the role of a clinical ethics fellowship. AB - BACKGROUND: Although clinical ethicists are becoming more prevalent in healthcare settings, their required training and education have not been clearly delineated. Most agree that training and education are important, but their nature and delivery remain topics of debate. One option is through completion of a clinical ethics fellowship. METHOD: In this paper, the first four fellows to complete a newly developed fellowship program discuss their experiences. They describe the goals, structure, participants and activities of the fellowship. They identify key elements for succeeding as a clinical ethicist and sustaining a clinical ethics program. They critically reflect upon the challenges faced in the program. RESULTS: The one-year fellowship provided real-time clinical opportunities that helped them to develop the necessary knowledge and skills, gain insight into the role and scope of practice of clinical ethicists and hone valuable character traits. CONCLUSION: The fellowship enabled each of the fellows to assume confidently and competently a position as a clinical ethicist upon completion. PMID- 15533245 TI - Sample size for detecting differentially expressed genes in microarray experiments. AB - BACKGROUND: Microarray experiments are often performed with a small number of biological replicates, resulting in low statistical power for detecting differentially expressed genes and concomitant high false positive rates. While increasing sample size can increase statistical power and decrease error rates, with too many samples, valuable resources are not used efficiently. The issue of how many replicates are required in a typical experimental system needs to be addressed. Of particular interest is the difference in required sample sizes for similar experiments in inbred vs. outbred populations (e.g. mouse and rat vs. human). RESULTS: We hypothesize that if all other factors (assay protocol, microarray platform, data pre-processing) were equal, fewer individuals would be needed for the same statistical power using inbred animals as opposed to unrelated human subjects, as genetic effects on gene expression will be removed in the inbred populations. We apply the same normalization algorithm and estimate the variance of gene expression for a variety of cDNA data sets (humans, inbred mice and rats) comparing two conditions. Using one sample, paired sample or two independent sample t-tests, we calculate the sample sizes required to detect a 1.5-, 2-, and 4-fold changes in expression level as a function of false positive rate, power and percentage of genes that have a standard deviation below a given percentile. CONCLUSIONS: Factors that affect power and sample size calculations include variability of the population, the desired detectable differences, the power to detect the differences, and an acceptable error rate. In addition, experimental design, technical variability and data pre-processing play a role in the power of the statistical tests in microarrays. We show that the number of samples required for detecting a 2-fold change with 90% probability and a p-value of 0.01 in humans is much larger than the number of samples commonly used in present day studies, and that far fewer individuals are needed for the same statistical power when using inbred animals rather than unrelated human subjects. PMID- 15533246 TI - Genomic structure and cloning of two transcript isoforms of human Sp8. AB - BACKGROUND: The Specificity proteins (Sp) are a family of transcription factors that have three highly conserved zinc-fingers located towards the carboxy terminal that bind GC-boxes and assist in the initiation of gene transcription. Human Sp1-7 genes have been characterized. Recently, the phenotype of Sp8 null mice has been described, being tailless and having severe truncation of both fore and hind limbs. They also have malformed brains with defective closure of the anterior and posterior neuropore during brain development. RESULTS: The human Sp8 gene is a three-exon gene that maps to 7p21.3, close to the related Sp4 gene. From an osteosarcoma cell line we cloned two transcript variants that use two different first exons and have a common second exon. One clone encodes a 508 residue protein, Sp8L (isoform 1) and the other a shorter 490-residue protein, Sp8S (isoform 2). These two isoforms are conserved being found also in mice and zebrafish. Analysis of the Sp8L protein sequence reveals an amino-terminal hydrophobic Sp-motif that is disrupted in Sp8S, a buttonhead box and three C2H2 zinc-fingers. Sp8 mRNA expression was detected in a wide range of tissues at a low level, with the highest levels being found in brain. Treatment of the murine pluripotent cell line C3H10T1/2 with 100 ng/mL BMP-2 induced Sp8 mRNA after 24 hours. CONCLUSIONS: There is conservation of the two Sp8 protein isoforms between primates, rodents and fish, suggesting that the isoforms have differing roles in gene regulation. Sp8 may play a role in chondrogenic/osteoblastic differentiation in addition to its role in brain and limb development. PMID- 15533247 TI - Functional characterization in Caenorhabditis elegans of transmembrane worm-human orthologs. AB - BACKGROUND: The complete genome sequences for human and the nematode Caenorhabditis elegans offer an opportunity to learn more about human gene function through functional characterization of orthologs in the worm. Based on a previous genome-wide analysis of worm-human orthologous transmembrane proteins, we selected seventeen genes to explore experimentally in C. elegans. These genes were selected on the basis that they all have high confidence candidate human orthologs and that their function is unknown. We first analyzed their phylogeny, membrane topology and domain organization. Then gene functions were studied experimentally in the worm by using RNA interference and transcriptional gfp reporter gene fusions. RESULTS: The experiments gave functional insights for twelve of the genes studied. For example, C36B1.12, the worm ortholog of three presenilin-like genes, was almost exclusively expressed in head neurons, suggesting an ancient conserved role important to neuronal function. We propose a new transmembrane topology for the presenilin-like protein family. sft-4, the worm ortholog of surfeit locus gene Surf-4, proved to be an essential gene required for development during the larval stages of the worm. R155.1, whose human ortholog is entirely uncharacterized, was implicated in body size control and other developmental processes. CONCLUSIONS: By combining bioinformatics and C. elegans experiments on orthologs, we provide functional insights on twelve previously uncharacterized human genes. PMID- 15533248 TI - Fibromatosis of the hand associated with EMO syndrome: a case report. AB - BACKGROUND: EMO syndrome, defined as a triad including exophthalmus, pretibial myxedema and osteoarthropathia, is a rare condition in patients suffering from hyperthyreosis. CASE PRESENTATION: We here describe an interesting case of EMO syndrome associated with unilateral fibromatosis of the hand and an initial stage of generalized myxedema of the skin. To our knowledge a similar case has not yet been described in literature though reports about associated fibromatosis, e.g. located retroperitoneally, already exist. Familiar explanations include its initiation by autoimmune processes or aberrant T-cell cytokine stimulation leading to an overwhelming production of glycosaminoglycans. CONCLUSION: Interpreting our case in context with previous reports we conclude that associated fibromatosis induced by autoimmune processes may affect a variety of different localizations and therefore requires careful monitoring. A therapeutical attempt by using UVA1 irridation for pretibial myxedema remained without a satisfying regression. PMID- 15533249 TI - "Any other comments?" Open questions on questionnaires - a bane or a bonus to research? AB - BACKGROUND: The habitual "any other comments" general open question at the end of structured questionnaires has the potential to increase response rates, elaborate responses to closed questions, and allow respondents to identify new issues not captured in the closed questions. However, we believe that many researchers have collected such data and failed to analyse or present it. DISCUSSION: General open questions at the end of structured questionnaires can present a problem because of their uncomfortable status of being strictly neither qualitative nor quantitative data, the consequent lack of clarity around how to analyse and report them, and the time and expertise needed to do so. We suggest that the value of these questions can be optimised if researchers start with a clear understanding of the type of data they wish to generate from such a question, and employ an appropriate strategy when designing the study. The intention can be to generate depth data or 'stories' from purposively defined groups of respondents for qualitative analysis, or to produce quantifiable data, representative of the population sampled, as a 'safety net' to identify issues which might complement the closed questions. SUMMARY: We encourage researchers to consider developing a more strategic use of general open questions at the end of structured questionnaires. This may optimise the quality of the data and the analysis, reduce dilemmas regarding whether and how to analyse such data, and result in a more ethical approach to making best use of the data which respondents kindly provide. PMID- 15533250 TI - Comparison of energy-restricted very low-carbohydrate and low-fat diets on weight loss and body composition in overweight men and women. AB - OBJECTIVE: To compare the effects of isocaloric, energy-restricted very low carbohydrate ketogenic (VLCK) and low-fat (LF) diets on weight loss, body composition, trunk fat mass, and resting energy expenditure (REE) in overweight/obese men and women. DESIGN: Randomized, balanced, two diet period clinical intervention study. Subjects were prescribed two energy-restricted (-500 kcal/day) diets: a VLCK diet with a goal to decrease carbohydrate levels below 10% of energy and induce ketosis and a LF diet with a goal similar to national recommendations (%carbohydrate:fat:protein = ~60:25:15%). SUBJECTS: 15 healthy, overweight/obese men (mean +/- s.e.m.: age 33.2 +/- 2.9 y, body mass 109.1 +/- 4.6 kg, body mass index 34.1 +/- 1.1 kg/m2) and 13 premenopausal women (age 34.0 +/- 2.4 y, body mass 76.3 +/- 3.6 kg, body mass index 29.6 +/- 1.1 kg/m2). MEASUREMENTS: Weight loss, body composition, trunk fat (by dual-energy X-ray absorptiometry), and resting energy expenditure (REE) were determined at baseline and after each diet intervention. Data were analyzed for between group differences considering the first diet phase only and within group differences considering the response to both diets within each person. RESULTS: Actual nutrient intakes from food records during the VLCK (%carbohydrate:fat:protein = ~9:63:28%) and the LF (~58:22:20%) were significantly different. Dietary energy was restricted, but was slightly higher during the VLCK (1855 kcal/day) compared to the LF (1562 kcal/day) diet for men. Both between and within group comparisons revealed a distinct advantage of a VLCK over a LF diet for weight loss, total fat loss, and trunk fat loss for men (despite significantly greater energy intake). The majority of women also responded more favorably to the VLCK diet, especially in terms of trunk fat loss. The greater reduction in trunk fat was not merely due to the greater total fat loss, because the ratio of trunk fat/total fat was also significantly reduced during the VLCK diet in men and women. Absolute REE (kcal/day) was decreased with both diets as expected, but REE expressed relative to body mass (kcal/kg), was better maintained on the VLCK diet for men only. Individual responses clearly show the majority of men and women experience greater weight and fat loss on a VLCK than a LF diet. CONCLUSION: This study shows a clear benefit of a VLCK over LF diet for short-term body weight and fat loss, especially in men. A preferential loss of fat in the trunk region with a VLCK diet is novel and potentially clinically significant but requires further validation. These data provide additional support for the concept of metabolic advantage with diets representing extremes in macronutrient distribution. PMID- 15533251 TI - The role of thallium-201 and pentavalent dimercaptosuccinic acid for staging cartilaginous tumours. AB - INTRODUCTION: Heterogeneity of cartilage tumours may confound accurate diagnosis and grading resulting in under and over treatment. Improved preoperative assessment of malignancy and grade would be invaluable for developing a rational plan for treatment. We examined correlations between nuclear tracer avidity and malignancy grade in cartilage tumours. METHODS: Between 1996 and 2000, 92 consecutive patients with cartilaginous tumours (50 benign, 42 non-metastatic malignant) underwent nuclear scanning. Thallium-201 (TL-201) and pentavalent dimercaptosuccinic acid (DMSAV) were used as nuclear isotopes. Scanning with these agents was performed on separate days 48 hours apart. Static and SPECT images were obtained at 30 m and 4 h after injection of nuclear tracer. Pathology review was undertaken blinded to the results of the nuclear scans and correlations between histologic results and trace uptake at 4 hours examined. RESULTS: 25 patients with negative DMSAV had benign tumours. 15/17 tumours with positive TL-201 had malignant tumours. 11/13 patients with both positive DMSAV and TL-201 scans had intermediate or high grade tumours and 4 of these developed metastases. We have developed an algorithm for the management of patients with tumours that aims to avoid over treatment of low grade tumours and under treatment of high grade tumours. CONCLUSION: Functional nuclear scanning with TL 201 and DMSAV complements other imaging modalities in the management of cartilaginous tumours. PMID- 15533252 TI - Results of a planned interim toxicity analysis with trimodality therapy, including carboplatin AUC = 4, paclitaxel, 5-fluorouracil, amifostine, and radiation for locally advanced esophageal cancer: preliminary analyses and treatment recommendations from the North Central Cancer Treatment Group. AB - PURPOSE: An aggressive trimodality approach from the Minnie Pearl Cancer Research Network [carboplatin AUC = 6, days 1 and 22; 5-fluorouracil 225 mg/m2 continuous infusion, days 1-42, paclitaxel 200 mg/m2, days 1 and 22; 45 Gy] has resulted in remarkable pathologic response rates but notable toxicity. This trial was designed to mitigate this toxicity by starting with a lower carboplatin dose, AUC = 4, and by adding subcutaneous amifostine. METHODS: This phase II trial included patients with locally advanced, potentially resectable esophageal cancer. All were to receive the above regimen with modifications of carboplatin AUC = 4 and amifostine 500 mg subcutaneously before radiation. All were then to undergo an esophagectomy. A planned interim toxicity analysis after the first 10 patients was to determine whether the carboplatin dose should escalate to AUC = 6. RESULTS: Ten patients were enrolled, and all required dose reductions/omissions during neoadjuvant therapy. One patient died from paclitaxel anaphylaxis. Six patients manifested a complete pathologic response. CONCLUSION: With this regimen, carboplatin AUC = 4 for patients with locally advanced esophageal cancer is appropriate. PMID- 15533253 TI - Visualization and exploratory analysis of epidemiologic data using a novel space time information system. AB - BACKGROUND: Recent years have seen an expansion in the use of Geographic Information Systems (GIS) in environmental health research. In this field GIS can be used to detect disease clustering, to analyze access to hospital emergency care, to predict environmental outbreaks, and to estimate exposure to toxic compounds. Despite these advances the inability of GIS to properly handle temporal information is increasingly recognised as a significant constraint. The effective representation and visualization of both spatial and temporal dimensions therefore is expected to significantly enhance our ability to undertake environmental health research using time-referenced geospatial data. Especially for diseases with long latency periods (such as cancer) the ability to represent, quantify and model individual exposure through time is a critical component of risk estimation. In response to this need a STIS - a Space Time Information System has been developed to visualize and analyze objects simultaneously through space and time. RESULTS: In this paper we present a "first use" of a STIS in a case-control study of the relationship between arsenic exposure and bladder cancer in south eastern Michigan. Individual arsenic exposure is reconstructed by incorporating spatiotemporal data including residential mobility and drinking water habits. The unique contribution of the STIS is its ability to visualize and analyze residential histories over different temporal scales. Participant information is viewed and statistically analyzed using dynamic views in which values of an attribute change through time. These views include tables, graphs (such as histograms and scatterplots), and maps. In addition, these views can be linked and synchronized for complex data exploration using cartographic brushing, statistical brushing, and animation. CONCLUSION: The STIS provides new and powerful ways to visualize and analyze how individual exposure and associated environmental variables change through time. We expect to see innovative space-time methods being utilized in future environmental health research now that the successful "first use" of a STIS in exposure reconstruction has been accomplished. PMID- 15533254 TI - Occurrence of symptoms and depressive mood among working-aged coronary heart disease patients. AB - BACKGROUND: The typical symptoms of coronary heart disease (CHD), chest pain and breathlessness, are well-known. They are considered quite dramatic, and can thus be fairly reliably mapped by a survey. However, people might have other clearly unpleasant symptoms impairing quality of life. The aim of this study is to evaluate the appearance of these complaints of working-aged people with self reported CHD. METHODS: The study consists of a postal questionnaire of randomly selected Finns in age groups 30-34, 40-44 and 50-54, a response rate of 39% (N = 15,477). The subjects were asked whether or not a doctor had told them that they had angina pectoris or had had myocardial infarction. Four randomly selected age and sex matched controls were chosen for every patient. The occurrence of self reported dyspnoea, chest pain during anger or other kind of emotion, palpitation and perspiration without physical exercise, irregular heartbeats, flushing, trembling of hands and voice, jerking of muscles, depression and day-time sleepiness were examined. Odds ratios (OR) with 95% confidence intervals (CI), between occurrence of symptoms and CHD with and without heart infarction, were computed by multivariate logistic regression analysis. RESULTS: The sample eventually comprised 319 CHD patients. Dyspnoea, chest pain during anger or other kind of emotion, palpitation, perspiration without physical exercise, irregular heartbeats daily or almost daily, trembling of hands and voice, and jerking of muscles occurred statistically significantly more frequently among CHD patients than among controls. The CHD patients also reported more depressive mood according to Beck's inventory scores and poorer sleep and more frequent day-time sleepiness than controls. In the multivariate logistic regression analysis chest pain during anger or other kind of emotion (ORs 4.12 and 3.61) and dyspnoea (ORs 2.33 and 3.81) were the symptoms most associated with CHD. CONCLUSIONS: Working aged people with self-reported coronary heart disease evince a number of symptoms limiting the quality of their every day life. This aspect should be paid attention to when evaluating functional capacity of these patients. PMID- 15533255 TI - Sick leave among home-care personnel: a longitudinal study of risk factors. AB - BACKGROUND: Sick leave due to neck, shoulder and back disorders (NSBD) is higher among health-care workers, especially nursing aides/assistant nurses, compared with employees in other occupations. More information is needed about predictors of sick leave among health care workers. The aim of the study was to assess whether self-reported factors related to health, work and leisure time could predict: 1) future certified sick leave due to any cause, in nursing aides/assistant nurses (Study group I) and 2) future self-reported sick leave due to NSBD in nursing aides/assistant nurses (Study group II). METHODS: Study group I, comprised 443 female nursing aides/assistant nurses, not on sick leave at baseline when a questionnaire was completed. Data on certified sick leave were collected after 18 months. Study group II comprised 274 of the women, who at baseline reported no sick leave during the preceding year due to NSBD and who participated at the 18 month follow-up. Data on sick leave due to NSBD were collected from the questionnaire at 18 months. The associations between future sick leave and factors related to health, work and leisure time were tested by logistic regression analyses. RESULTS: Health-related factors such as previous low back disorders (OR: 1.89; 95% CI 1.20-2.97) and previous sick leave (OR 6.40; 95%CI 3.97-10.31), were associated with a higher risk of future sick leave due to any cause. Factors related to health, work and leisure time, i.e. previous low back disorders (OR: 4.45; 95% CI 1.27-15.77) previous sick leave, not due to NSBD (OR 3.30; 95%CI 1.33-8.17), high strain work (OR 2.34; 95%CI 1.05-5.23) and high perceived physical exertion in domestic work (OR 2.56; 95%CI 1.12-5.86) were associated with a higher risk of future sick leave due to NSBD. In the final analyses, previous low back disorders and previous sick leave remained significant in both study groups. CONCLUSION: The results suggest a focus on previous low back disorders and previous sick leave for the design of early prevention programmes aiming at reducing future sick leave due to any cause, as well as due to NSBD, among nursing aides/assistant nurses. A multifactorial approach may be of importance in the early prevention of sick leave due to NSBD. PMID- 15533256 TI - A randomised controlled trial to determine the effect on response of including a lottery incentive in health surveys [ISRCTN32203485]. AB - BACKGROUND: Postal questionnaires are an economical and simple method of data collection for research purposes but are subject to non-response bias. Several studies have explored the effect of monetary and non-monetary incentives on response. Recent meta-analyses conclude that financial incentives are an effective way of increasing response rates. However, large surveys rarely have the resources to reward individual participants. Three previous papers report on the effectiveness of lottery incentives with contradictory results. This study aimed to determine the effect of including a lottery-style incentive on response rates to a postal health survey. METHODS: Randomised controlled trial. SETTING: North and West Birmingham. 8,645 patients aged 18 or over randomly selected from registers of eight general practices (family physician practices). INTERVENTION: Inclusion of a flyer and letter with a health questionnaire informing patients that returned questionnaires would be entered into a lottery-style draw for pound 100 of gift vouchers. CONTROL: Health questionnaire accompanied only by standard letter of explanation. MAIN OUTCOME MEASURES: Response rate and completion rate to questionnaire. RESULTS: 5,209 individuals responded with identical rates in both groups (62.1%). Practice, patient age, sex and Townsend score (a postcode based deprivation measure) were identified as predictive of response, with higher response related to older age, being female and living in an area with a lower Townsend score (less deprived). CONCLUSION: This RCT, using a large community based sample, found that the offer of entry into a lottery style draw for pound 100 of High Street vouchers has no effect on response rates to a postal health questionnaire. PMID- 15533257 TI - Case-based medical informatics. AB - BACKGROUND: The "applied" nature distinguishes applied sciences from theoretical sciences. To emphasize this distinction, we begin with a general, meta-level overview of the scientific endeavor. We introduce the knowledge spectrum and four interconnected modalities of knowledge. In addition to the traditional differentiation between implicit and explicit knowledge we outline the concepts of general and individual knowledge. We connect general knowledge with the "frame problem," a fundamental issue of artificial intelligence, and individual knowledge with another important paradigm of artificial intelligence, case-based reasoning, a method of individual knowledge processing that aims at solving new problems based on the solutions to similar past problems. We outline the fundamental differences between Medical Informatics and theoretical sciences and propose that Medical Informatics research should advance individual knowledge processing (case-based reasoning) and that natural language processing research is an important step towards this goal that may have ethical implications for patient-centered health medicine. DISCUSSION: We focus on fundamental aspects of decision-making, which connect human expertise with individual knowledge processing. We continue with a knowledge spectrum perspective on biomedical knowledge and conclude that case-based reasoning is the paradigm that can advance towards personalized healthcare and that can enable the education of patients and providers. We center the discussion on formal methods of knowledge representation around the frame problem. We propose a context-dependent view on the notion of "meaning" and advocate the need for case-based reasoning research and natural language processing. In the context of memory based knowledge processing, pattern recognition, comparison and analogy-making, we conclude that while humans seem to naturally support the case-based reasoning paradigm (memory of past experiences of problem-solving and powerful case matching mechanisms), technical solutions are challenging.Finally, we discuss the major challenges for a technical solution: case record comprehensiveness, organization of information on similarity principles, development of pattern recognition and solving ethical issues. SUMMARY: Medical Informatics is an applied science that should be committed to advancing patient-centered medicine through individual knowledge processing. Case-based reasoning is the technical solution that enables a continuous individual knowledge processing and could be applied providing that challenges and ethical issues arising are addressed appropriately. PMID- 15533259 TI - Overweight, obesity, and colorectal cancer screening: disparity between men and women. AB - BACKGROUND: To estimate the association between body-mass index (BMI: kg/m2) and colorectal cancer (CRC) screening among US adults aged >or= 50 years. METHODS: Population-based data from the 2001 Behavioral Risk Factor Surveillance Survey. Adults (N = 84,284) aged >or= 50 years were classified by BMI as normal weight (18.5-<25), overweight (25-<30), obesity class I (30-<35), obesity class II (35 <40), and obesity class III (>or= 40). Interval since most recent screening fecal occult blood test (FOBT): (0 = >1 year since last screening vs. 1 = screened within the past year), and screening sigmoidoscopy (SIG): (0 = > 5 years since last screening vs. 1 = within the past 5 years) were the outcomes. RESULTS: Results differed between men and women. After adjusting for age, health insurance, race, and smoking, we found that, compared to normal weight men, men in the overweight (odds ratio [OR] 1.25, 95% CI = 1.05-1.51) and obesity class I (OR = 1.21, 95% CI = 1.03-1.75) categories were more likely to have obtained a screening SIG within the previous 5 years, while women in the obesity class I (OR = 0.86, 95%CI = 0.78-0.94) and II (OR = 0.88, 95%CI = 0.79-0.99) categories were less likely to have obtained a screening SIG compared to normal weight women. BMI was not associated with FOBT. CONCLUSION: Weight may be a correlate of CRC screening behavior but in a different way between men and women. PMID- 15533258 TI - Morphological correlates of injury-induced reorganization in primate somatosensory cortex. AB - BACKGROUND: Topographic reorganization of central maps following peripheral nerve injury has been well characterized. Despite extensive documentation of these physiological changes, the underlying anatomical correlates have yet to be fully explored. In this study, we used Golgi impregnation and light microscopy to assess dendritic morphology following denervation of the glabrous hand surface in adult primates. RESULTS: After survival durations that permit complete physiologically-defined reorganization, we find a systematic change in the dendritic arborization pattern of both layer II/III pyramidal and layer IV spiny stellate cells in the contralateral hand region of area 3b, compared to unaffected cortical areas. In general, our analyses indicate a progressive expansion of distal regions of the dendritic arbor with no appreciable changes proximally. This pattern of distal dendritic elaboration occurs for both basilar and apical dendrites. CONCLUSIONS: These observations are consistent with the notion that latent inputs gain expression in reorganized cortex after nerve injury via their influence through contacts with more distally located termination sites. PMID- 15533260 TI - The Chernobyl childhood leukemia study: background & lessons learned. AB - Many challenges emerged during completion of a study to examine radiation dose and acute leukemia among children in areas of the former Soviet Union. In an era of globalization, our experiences might benefit others involved in multinational investigations. PMID- 15533261 TI - Dietary effect of pomegranate seed oil rich in 9cis, 11trans, 13cis conjugated linolenic acid on lipid metabolism in obese, hyperlipidemic OLETF rats. AB - Conjugated fatty acid, the general term of positional and geometric isomers of polyunsaturated fatty acids with conjugated double bonds, has attracted considerable attention because of its potentially beneficial biological effects. In the present study, dietary effect of pomegranate seed oil rich in punicic acid (9cis, 11trans, 13cis-conjugated linolenic acid; 9c, 11t, 13c-CLNA) on lipid metabolism was investigated in obese, hyperlipidemic Otsuka Long-Evans Tokushima Fatty (OLETF) rats. After 2 weeks feeding period, OLETF rats revealed obesity and hyperlipidemia compared with their progenitor LETO rats. Feeding of the diet supplemented with 9% safflower oil and 1% pomegranate seed oil (9c, 11t, 13c-CLNA diet) did not affect abdominal white adipose tissue weights and serum lipid levels compared with the diet supplemented with 10% safflower oil (control diet) in OLETF rats. However, the accumulated hepatic triacylglycerol was markedly decreased by 9c, 11t, 13c-CLNA diet in OLETF rats. Activities of hepatic enzymes related to fatty acid synthesis and fatty acid beta-oxidation were not altered by 9c, 11t, 13c-CLNA diet. Levels of monounsaturated fatty acid (MUFA), major storage form of fatty acid, in serum triacylglycerol were markedly higher in obese, hyperlipidemic OLETF rats than in lean LETO rats. In addition, 9c, 11t, 13c-CLNA diet significantly decreased MUFA levels in OLETF rats. This is the first study showing that 9c, 11t, 13c-CLNA suppresses delta-9 desaturation in vivo, and we suggest that the alleviation of hepatic triacylglycerol accumulation by 9c, 11t, 13c-CLNA diet was, at least in part, attributable to the suppression of delta-9 desaturation in OLETF rats. PMID- 15533262 TI - Various non-digestible saccharides increase intracellular calcium ion concentration in rat small-intestinal enterocytes. AB - We have previously shown that non-digestible saccharides (NDS) stimulate intestinal Ca absorption via tight junctions. However, the cellular mechanisms activated by the NDS are not yet known. We investigated the effects of four NDS, difructose anhydride (DFA) III, DFAIV, fructo-oligosaccharides, and maltitol, on intracellular Ca signalling in isolated rat small-intestinal enterocytes. The changes in intracellular Ca(2+) concentration were measured before and after the addition of capric acid (7.5 or 15 mmol/l, a positive control), glycerol, or each NDS (1 or 10 mmol/l) to fura-2-loaded enterocytes. Treatment with capric acid or each NDS caused an immediate and dose-dependent rise in intracellular Ca(2+) concentration. Mechanical and osmotic stimulation achieved by adding glycerol had no effect on intracellular Ca(2+) concentration. The intracellular Ca(2+) concentration in enterocytes treated with DFAIII and fructo-oligosaccharides reached a peak level at about 30 s after stimulation, but those treated with DFAIV and maltitol showed further increases after the initial rapid rise. The maximum change in intracellular Ca(2+) concentration obtained by the application of maltitol was higher than that of DFAIII at 10 mmol/l. These findings suggest that each of the four NDS directly stimulates rat enterocytes, and increases intracellular Ca(2+) concentration. Thus, molecular structure may be more important than the size of the NDS in the induction of Ca signalling in the cells. PMID- 15533263 TI - Potential modulation of plasma ghrelin and glucagon-like peptide-1 by anorexigenic cannabinoid compounds, SR141716A (rimonabant) and oleoylethanolamide. AB - The CB1 cannabinoid receptor antagonist, N-piperidino-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methylpyrazole-3-carboxamide (rimonabant; SR141716A), and oleoylethanolamide (OEA) are known to reduce food consumption, by, at least partially, a peripheral regulation of feeding. The effects of systemic SR141716A or OEA (5 mg/kg) administrations on food consumption in 24 h food-deprived and fed rats were investigated. In fasted rats, SR141716A and OEA produced an inhibition in food intake measurable the first 20 min following injection. The increase in ghrelin levels observed in the vehicle-injected rats was abolished in animals receiving OEA and significantly reduced with SR141716A. Neither OEA nor SR141716A modified glucagon-like peptide-1 (7-36) amide portal levels 20 min after the administration. In fed rats, plasma ghrelin levels of SR141716A- and OEA-treated rats were 35% lower as compared with those of the vehicle-injected rats. These results show an influence of cannabinoid agents on circulating ghrelin levels and suggest that their short-term action on appetite seems to be in accordance with the control of secretion of gastrointestinal orexigenic peptides, mainly expressed in the upper part of the gastrointestinal tract. PMID- 15533264 TI - Response of ApoA-IV in pigs to long-term increased dietary oil intake and to the degree of unsaturation of the fatty acids. AB - ApoA-IV is a protein constituent of HDL particles; the gene coding for it is a member of the ApoA-I-ApoC-III-ApoA-IV cluster. To investigate the effects of the quantity and the degree of saturation of dietary lipid on the long-term response of this Apo, and on the hypothetical coordinated regulation of the cluster in vivo, pigs were fed isoenergetic, cholesterol-free, low-lipid or lipid-enriched diets (containing either extra olive oil (rich in MUFA) or sunflower oil (rich in n-6 PUFA)) for 42 d. In animals fed on the control diet, ApoA-IV was mainly associated with plasma lipoproteins. An increase in plasma ApoA-IV concentration, mainly in the lipoprotein-free fraction, was induced by the lipid-enriched diets, independent of the degree of saturation of the fatty acids involved. The latter diets also led to increases in hepatic ApoA-I, ApoA-IV and ApoC-III mRNA levels, more so with the sunflower oil-rich diet. The present results show that porcine plasma ApoA-IV levels and their association with lipoproteins are very sensitive to increases in dietary lipids, independent of the degree of fatty acid saturation. Furthermore, hepatic expression of RNA appears to be coordinated along with that of the other members of the gene cluster. PMID- 15533265 TI - Lactobacillus gasseri: effects on mouse intestinal flora enzyme activity and isoflavonoids in the caecum and plasma. AB - The effects of Lactobacillus gasseri JCM 1131(T) on isoflavonoid levels within the caecum and plasma were assessed in adult mice. Male 5-week-old mice were fed an AIN 93M diet for 30 d. Two groups of mice were administered either L. gasseri JCM 1131(T) (the LGI group) or physiological saline solution (the control (CI) group) daily for 5 d before dissection. The plasma daidzein concentration was significantly higher in the LGI group, however, their plasma equol concentration was significantly less than in the CI group. The total amount of equol present as aglycone in the caecum was significantly greater in the CI group, but there was no significant difference in the total daidzein present as caecal aglycone. In an in vitro incubation of daidzein with the faecal flora of mice, the equol concentration was significantly higher in the CI group. The numbers of lactobacilli present were significantly higher in the LGI group. The present data suggest that the administration of L. gasseri is likely to influence the effect of isoflavonoids on the host via changes in the gastrointestinal environment. PMID- 15533266 TI - Apical sodium-glucose co-transport can be regulated by blood-borne glucose in the ruminal epithelium of sheep (Ovis aries, Merino breed). AB - The intestinal Na-dependent D-glucose co-transporter (SGLT)-1 in sheep is under dietary regulation by luminal substrates. The aim of the present study was to find out whether the SGLT-1 in the forestomach of sheep is also regulated by sugars. Furthermore, the location of a possible glucosensor (luminal v. intracellular v. basolateral) was to be elucidated. Ruminal epithelia of sheep (Ovis aries, Merino breed) were pre-incubated in Ussing chambers with various substrates on the mucosal (i.e. luminal) or serosal (i.e. blood) side. This pre incubation period was followed by a second pre-incubation period without the tested substrates (washout period). Thereafter, apical D-glucose uptake by ruminal epithelial cells was determined with 200 mumol D-[(14)C]glucose/l in the absence or co-presence of the SGLT-1 inhibitor, phlorizin. Pre-incubation with D glucose on the mucosal side had no significant effect on apical D-glucose uptake (P>0.05). In contrast, pre-incubation with D-glucose, D-mannose, 3-O-methyl-d glucose or sucrose on the serosal side significantly increased D-glucose uptake compared with mannitol-treated controls (P<0.05). Serosal pre-incubation with cellobiose or D-xylose had no effect. The stimulation of d-glucose uptake by serosal D-glucose pre-incubation was concentration dependent, with maximal stimulation at about 10 mmol/l. We conclude that the ruminal SGLT-1 can be up regulated in a concentration-dependent manner by blood-borne D-glucose via an extracellular sugar-sensing mechanism. PMID- 15533267 TI - Hypoglycaemic and anorexigenic activities of an alpha-amylase inhibitor from white kidney beans (Phaseolus vulgaris) in Wistar rats. AB - An inhibitor of alpha-amylase was isolated and purified from an extract of white kidney beans (Phaseolus vulgaris). The acute oral administration of the inhibitor (50 mg/kg body weight) to adult Wistar rats together with a starch load (2 g/kg body weight suspended in NaCl (9 g/l)) reduced the increase in glycaemia over the basal value (NaCl, 222 (SEM 49); inhibitor, 145 (SEM 16) mmol/l x 180 min; P<0.05) without modifying the insulin response. On administering the inhibitor orally (50 mg/kg body weight dissolved in NaCl (9 g/l)) for 21 d to rats fed on a standard diet, a decline was observed in the glycaemia values on day 0 (NaCl, 5.53 (SEM 0.12); inhibitor, 5.25 (SEM 0.16) mmol/l) relative to those obtained on days 10 (NaCl, 5.00 (SEM 0.14); inhibitor, 4.60 (SEM 0.08) mmol/l; P<0.05) and 21 (NaCl, 5.22 (SEM 0.22); inhibitor, 4.50 (SEM 0.12) mmol/l; P<0.01) of treatment, without modifying the plasma concentration of insulin. There was found to be a significant anorexigenic action of the inhibitor; there was reduced food intake (NaCl, 23.07 (SEM 0.31); inhibitor, 19.50 (SEM 0.49) g/d; P<0.01), a reduced weight gain (NaCl, 52 (SEM 3); inhibitor, -1.33 (SEM 8.9) g/21 d; P<0.01), as well as changes in the activity of some intestinal enzymes such as maltase (NaCl, 87 (SEM 7); inhibitor, 127 (SEM 11) U/g proteins; P<0.05). The present study has shown, for the first time, that the prolonged administration of an alpha-amylase inhibitor reduces blood glucose levels and body-weight gain in Wistar rats. PMID- 15533268 TI - A comparison of the effects of cheese and butter on serum lipids, haemostatic variables and homocysteine. AB - Milk fat contains considerable amounts of saturated fatty acids, known to increase serum cholesterol. Little is known, however, about the relative effect of different milk products on risk factors for CHD. The aim of the present study was to compare the effects of Jarlsberg cheese (a Norwegian variety of Swiss cheese) with butter on serum lipoproteins, haemostatic variables and homocysteine. A controlled dietary study was performed with twenty-two test individuals (nine men and thirteen women) aged 23-54 years. The subjects consumed three isoenergetic test diets, with equal amounts of fat and protein, and containing either cheese (CH diet), butter + calcium caseinate (BC diet) or butter + egg-white protein (BE diet). The study was a randomised cross-over study and the subjects consumed each diet for 3 weeks, with 1 week when they consumed their habitual diet in between. Fasting blood samples were drawn at baseline and at the end of each period. Serum was analysed for lipids and plasma for haemostatic variables and homocysteine. Total cholesterol was significantly lower after the CH diet than after the BC diet (-0.27 mmol/l; P=0.03), while the difference in LDL-cholesterol was found to be below significance level (-0.22 mmol/l; P=0.06). There were no significant differences in HDL-cholesterol, triacylglycerols, apo A-I, apo B or lipoprotein (a), haemostatic variables and homocysteine between the diets. The results indicate that, at equal fat content, cheese may be less cholesterol increasing than butter. PMID- 15533269 TI - Malnutrition in hospital outpatients and inpatients: prevalence, concurrent validity and ease of use of the 'malnutrition universal screening tool' ('MUST') for adults. AB - The 'malnutrition universal screening tool' ('MUST') for adults has been developed for all health care settings and patient groups, but ease of use and agreement with other published tools when screening to identify malnutrition requires investigation. The present study assessed the agreement and the prevalence of malnutrition risk between 'MUST' and a variety of other tools in the same patients and compared the ease of using these tools. Groups of patients were consecutively screened using 'MUST' and: (1) MEREC Bulletin (MEREC) and Hickson and Hill (HH) tools (fifty gastroenterology outpatients); (2) nutrition risk score (NRS) and malnutrition screening tool (MST; seventy-five medical inpatients); (3) short-form mini nutritional assessment (MNA-tool; eighty-six elderly and eighty-five surgical inpatients); (4) subjective global assessment (SGA; fifty medical inpatients); (5) Doyle undernutrition risk score (URS; fifty two surgical inpatients). Using 'MUST', the prevalence of malnutrition risk ranged from 19-60% in inpatients and 30% in outpatients. 'MUST' had 'excellent' agreement (kappa 0.775-0.893) with MEREC, NRS and SGA tools, 'fair-good' agreement (kappa 0.551-0.711) with HH, MST and MNA-tool tools and 'poor' agreement with the URS tool (kappa 0.255). When categorisation of malnutrition risk differed between tools, it did not do so systematically, except between 'MUST' and MNA-tool (P=0.0005) and URS (P=0.039). 'MUST' and MST were the easiest, quickest tools to complete (3-5 min). The present investigation suggested a high prevalence of malnutrition in hospital inpatients and outpatients (19-60% with 'MUST') and 'fair-good' to 'excellent' agreement beyond chance between 'MUST' and most other tools studied. 'MUST' was quick and easy to use in these patient groups. PMID- 15533270 TI - A substitution model of dietary manipulation is an effective means of optimising lipid profile, reducing C-reactive protein and increasing insulin-like growth factor-1. AB - There are two key methods in which fat intake may be manipulated; the 'substitution model' and the 'reduction model'. However insufficient information is known about the mechanisms of dietary fat reduction in individuals who have successfully reduced their fat intake, to be clear as to which strategy offers the greatest chance of success. Our objective was to ascertain the most effective dietary intervention for improving cardiovascular risk profile. Eighty female volunteers (high fat consumers) were recruited. Each subject was randomly allocated into one of the following groups. Substitution of high-fat foods was made with reduced-fat products, by the reduction of high-fat foods, by a combination of substitution and reduction strategies, or no advice was given. Each intervention lasted 3 months. Anthropometric measures and fasting blood samples were taken at baseline and follow-up. The substitution intervention resulted in weight loss (mean -1.4 (95 % CI -2.4, -0.2) kg) and reduced percentage body fat (mean -1.3 (95% CI -2.0, -0.5)%). There was no significant weight change with the other interventions. Fasting triacylglycerols (-0.2 (SEM 0.07) mm; P=0.04), cholesterol and C-reactive protein (CRP) levels (0.8 (SEM 0.2) mg/l; P=0.04) fell with the substitution intervention, but not with the other interventions. Insulin-like growth factor-1 increased with both substitution and reduction (P=0.02). There was no significant change in fasting insulin or glucose with any intervention. The substitution model of dietary intervention is effective even over a relatively short interval of time in reducing fasting total cholesterol, triacylglycerols and CRP. Although the group size for the present study was small and involved females only, it has significant implications for population intervention strategies. PMID- 15533271 TI - Stunted children gain less lean body mass and more fat mass than their non stunted counterparts: a prospective study. AB - The aim of the present study was to analyse the changes in body composition of stunted children during a follow-up period and to test the hypothesis of a tendency to accumulate body fat as a consequence of undernutrition early in life. We selected fifty boys and girls aged 11 to 15, who were residents of slums in Sao Paulo, Brazil. Twenty were stunted (S) and thirty had normal stature (NS). The children's nutritional status and body composition were assessed through anthropometry and dual-energy X-ray absorptiometry, at the beginning of the present study and after 3 years, and changes in lean mass (LM and LM%) and fat mass (FM and FM%) were calculated. Stunted boys accumulated more body fat (FM%: S=1.62%, NS=-3.40%; P=0.003) and gained less lean mass (LM%: S=-1.46, NS=3.21%; P=0.004). Stunted girls gained less lean mass (S=7.87 kg, NS=11.96 kg; P=0.032) and had significantly higher values of FM% at follow-up when compared with their baseline values (P=0.008), whereas non-stunted girls had a non-significant difference in FM% over time (P=0.386). These findings are important to understand the factors involved in the increased prevalence of overweight and obesity among poor populations, which appear to be associated with hunger during infancy and/or childhood. PMID- 15533272 TI - Comparison of predicted body fat percentage from anthropometric methods and from impedance in university students. AB - The objective of the present study was to compare different methods for evaluating body fat percentage (BF%) (anthropometric methods and bioelectrical impedance analysis) in university students. Subjects were 653 healthy students whose mean age, body height, body weight and BMI were 21.1 (SD 2.5) years, 166.0 (SD 8.4) cm, 62.8 (SD 11.0) kg and 22.7 (SD 3.1) kg/m(2), respectively. Results showed that BMI is a poor predictor of body fatness since the sensitivity was low in comparison with the reference method (Siri equation). The lowest values of BF% were obtained using the reference method (Siri equation) (21.8 (SD 6.8)%). The two methods with the highest agreement were Siri and Lean (mean difference, 0.5), followed by Brozek (mean difference, -1.4) and Deurenberg (mean difference, -1.5). The largest mean difference for BF% was between Siri and impedance (-4.5). Although the methods and/or equations used in the present study have been commonly utilised to estimate BF% in young adults, the results must be interpreted with caution in the diagnosis and monitoring of overweight and obesity. PMID- 15533273 TI - Maternal body composition, HIV infection and other predictors of gestation length and birth size in Zimbabwe. AB - The role of maternal infections, nutritional status and obstetric history in low birth weight is not clear. Thus, the objective of the present study was to assess the effects of maternal HIV infection, nutritional status and obstetric history, and season of birth on gestation length and birth size. The study population was 1669 antenatal care attendees in Harare, Zimbabwe. A prospective cohort study was conducted as part of a randomised, controlled trial. Maternal anthropometry, age, gravidity, and HIV status and load were assessed in 22nd-35th weeks gestation. Outcomes were gestation length and birth size. Birth data were available from 1106 (66.3%) women, of which 360 (32.5%) had HIV infection. Mean gestation length was 39.1 weeks with 16.6% <37 weeks, mean birth weight was 3030 g with 10.5% <2500 g. Gestation length increased with age in primigravidae, but not multigravidae (interaction, P=0.005), and birth in the early dry season, low arm fat area, multiple pregnancies and maternal HIV load were negative predictors. Birth weight increased with maternal height, and birth in the late rainy and early dry season; primi-secundigravidity, low arm fat area, HIV load, multiple pregnancies and female sex were negative predictors. In conclusion, gestation length and birth weight decline with increasing maternal HIV load. In addition, season of birth, gravidity, maternal height and body fat mass, and infant sex are predictors of birth weight. PMID- 15533274 TI - Energy restriction initiated at different gestational ages has varying effects on maternal weight gain and pregnancy outcome in common marmoset monkeys (Callithrix jacchus). AB - With relatively high fertility and short lifespan, marmoset monkeys (Callithrix jacchus) may become useful primate models of prenatal nutritional effects on birth condition and adult disease risk. The present study determined the effects of energy restriction to 75% of expected ad libitum consumption during mid- (day 66) or late (day 99) gestation on maternal weight, fetal growth and pregnancy outcomes in this species. Mid-restriction reliably induced the loss of pregnancy before term, at 92 d, on average. Of the late-restricted pregnancies, four of seven were normal term length while three were preterm deliveries, at 101, 117 and 132 d. Control females had a mean mid-pregnancy weight gain of 0.67 g/d while mid-restricted females lost -0.65 g/d, on average. Control pregnancies averaged a 1.06 g/d gain during late pregnancy, while energy-restricted females lost -0.67 g/d, on average. Restriction-related weight change was highly variable, ranging from +0.55 to -2.56 g/d for mid-restriction pregnancies and from +0.79 to -3.91 g/d for late-restriction pregnancies. For mid-restriction pregnancies, the number of restriction days was best explained by linear weight change and total weight loss while the number of restriction days in late pregnancy was best explained by linear weight change alone. In late-restriction pregnancies, smaller females had higher daily weight losses. Restrictions did not induce litter-size reduction or growth restriction in those infants that were delivered at term but the size of aborted fetuses suggested that at least some pregnancies lost preterm may have involved impaired intra-uterine growth. PMID- 15533275 TI - Validity of a food-frequency questionnaire for a large prospective cohort study in Bangladesh. AB - We have developed a thirty-nine-item semi-quantitative food-frequency questionnaire (FFQ) to assess the dietary consumption of 11,746 men and women in a prospective cohort study that evaluates the health effects of As from drinking water in Bangladesh. In order to validate the FFQ, two 7 d food diaries (FD) were completed for 189 randomly selected cohort participants in two different seasons of the year. Nutrient values were converted based on both the United States Department of Agriculture's National Nutrient Database and a food composition table for the Indian subcontinent. Pearson product-moment and Spearman non parametric rank correlation coefficients comparing food and nutrient consumptions estimated from FFQ and 7 d FD were calculated based on log-transformed consumption values with or without adjustment for total energy and correction for within-individual variation. Correlations of macronutrients and common micronutrients including total fat, monounsaturated fat, polyunsaturated fat, saturated fat, protein, carbohydrate, dietary fibre, Na, K, vitamin B6, vitamin B12, riboflavin, Mn, thiamin and Fe were moderately good, ranging from 0.30 to 0.76. However, correlations of other micronutrients were weak (<0.30). Large seasonal variations in intakes of retinol equivalents and vitamin C were observed. This analysis documents the degree of validity of the FFQ in measuring specific nutrient intakes in the study population. To our knowledge, the present study is the first to document the validity of a FFQ with the use of 7 d FD in a Bangladeshi population. PMID- 15533276 TI - Oral candidiasis and nutritional deficiencies in elderly hospitalised patients. AB - The prevalence of oral candidiasis and its association with malnutrition in terms of protein-energy malnutrition and mineral and vitamin depletion were evaluated in ninety-seven hospitalised older adults aged 82.1 (SD 8.6) years. Patients underwent a complete oral examination with microbiological investigation on admission to our geriatric rehabilitation unit. Patients were assessed nutritionally by evaluation of dietary intake and measurement of anthropometric variables, serum nutritional proteins, ferritin, Zn, folate, vitamins B12 and C. The prevalence of oral candidiasis was 37% (n 36); the proportion of patients with BMI <20 kg/m(2) was 32% (n 31). The nutritional status of the population was studied by comparing two groups defined according to the absence (group I; n 61) or presence (group II; n 36) of oral candidiasis. The two groups did not differ on the basis of BMI and mid-arm circumference. However, group II had a smaller leg circumference, lower daily energy and protein intakes, lower albumin and transthyretin levels. Patients successfully treated with fluconazole increased their intake on day 30. The proportion of patients with hypozincaemia (<12.5 micromol/l) and vitamin C deficiency (<0.7 mg/l) was higher in group II. Treatment with antibiotics, poor oral hygiene, denture wearing, and vitamin C deficiency appeared as the most significant independent risk factors associated with oral candidiasis. The present findings show that oral candidiasis appears to be related to malnutrition and results in mucosal lesions that have a negative impact on energy intake, which may subsequently worsen nutritional status. PMID- 15533277 TI - Comparison of FNAC and open biopsy in palpable breast lumps. AB - OBJECTIVE: To compare the results of FNAC and open biopsy in patients presented with palpable breast lump. DESIGN: Comparative study. PLACE AND DURATION OF STUDY: Nishtar Hospital, Multan, during the period of October 2001 to October 2003. PATIENTS AND METHODS: All female patients, irrespective of their age, who presented with breast lump were included in the study. The patients were divided into two groups, group I & II of clinically benign and clinically malignant respectively. Both groups underwent FNAC. The patients with suspicious FNAC were subjected to excision biopsy in group I patients. Group II patients were advised surgery and final report was made on histopathology. Sensitivity and specificity of the FNAC was determined. RESULTS: A total of 89 cases were included in whom both FNAC and histopathology results were available for comparison. Clinically, 54 were benign and 35 were malignant. The cytological diagnosis was unequivocally malignant in 29 patients, suspicious in 3 cases, unequivocally benign in 44 patients and probably benign in 7 patients while specimen was inadequate in 6 patients. Histological diagnosis of these 89 patients showed 55 patients with benign disease and 34 patients having malignant disease. In malignant disease, sensitivity of the FNAC was 85.29% with 100% specificity, 14.7% false negative rate, 100% positive predictive value and 98.79% negative predictive value. CONCLUSION: FNAC has good sensitivity (85.29%) and very high specificity (100%). It can replace the open biopsy in majority cases of clinically malignant disease. Although FNAC is slightly less sensitive (80%) in benign diseases, it is highly specific (100%), so it can help to reassure and relieve the anxiety of the patients. PMID- 15533278 TI - Childhood nocturnal enuresis: developing a conceptual framework. AB - Nocturnal enuresis has been described as the most prevalent and chronic of all childhood problems. The experience, particularly for the older child, can be extremely distressing and limiting. It is now thought that psychological distress (emotional, behavioural, and self-esteem) arises as a consequence of bedwetting, and reaches clinical importance in only a minority who are vulnerable. Many aetiological theories have been proposed with the cause of nocturnal enuresis now regarded as heterogeneous. Based on empirical findings, a new model, termed 'the three systems,' has facilitated a greater clinical understanding of the problem and identification of the appropriate intervention. The model proposes bedwetting results from excessive nocturnal urine production and/or nocturnal bladder overactivity coupled with an inability to arouse to bladder sensations. Effective treatment arises from identification of the child's particular need and application of the appropriate psychological and pharmacological approach. Treatment methods are reviewed in terms of the new model, the mode of action, effectiveness, and application in combination. PMID- 15533279 TI - Is premenstrual dysphoria a variant of panic disorder? A review. AB - Patients with premenstrual dysphoric disorder (PMDD) and patients with panic disorder (PD) both experience high rates of panic attacks in laboratory panic provocation studies. Recently, this shared elevated rate of challenge-induced panic has received increasing attention. Researchers have suggested that PMDD and panic disorder may share a pathophysiological or psychobiological link. The purpose of this paper is to review the findings from PMDD challenge studies and the theories advanced to connect PMDD to panic disorder. Taken together, the results of the PMDD challenge studies confirm that agents that incite panic in PD patients do so as well in PMDD women. This shared elevated challenge-induced panic cannot be accounted for by explanations such as a history of PD in PMDD women. None of the physiological theories as currently expressed--suffocation false alarm, gamma-aminobutyric acid (GABA), noradrenergic, serotonergic, and cholecystokinin--yet provides a compelling candidate to account for shared elevated challenge-induced panic in PD and PMDD patients. Psychological perspectives on panic emphasize that bodily sensations themselves can cause fear. Researchers have yet to apply several influential psychological approaches- conditioning, catastrophic misinterpretation, and anxiety sensitivity--to PMDD patients. Because psychological factors influence anxious responding in challenge studies, the search for the biological abnormality best accounting for PMDD panic might benefit from a reframing of the question to one that considers the psychological perspective as well. PMID- 15533280 TI - Factors associated with suicidal phenomena in adolescents: a systematic review of population-based studies. AB - Suicidal phenomena (suicide attempts, deliberate self-harm, and suicidal plans, threats and thoughts) are common in adolescents. Identification of factors associated with these phenomena could play an important role in the development of school or community-based prevention and intervention programs. In this article, we report the results of a systematic review of the international literature on population-based studies of factors associated with suicidal phenomena in adolescents. These factors encompass psychiatric, psychological, physical, personal, familial and social domains. The quantity of evidence in support of associations between suicidal phenomena and specific factors is compared with the quantity of evidence against such associations. We conclude with a summary of the findings, including identification of new or neglected areas, which require further investigation. Methodological considerations are highlighted and implications of the findings for clinicians and other professionals concerned with prevention of suicidal behavior by adolescents are discussed. PMID- 15533281 TI - Gender differences in risk factors and consequences for alcohol use and problems. AB - Women drink less alcohol and have fewer alcohol-related problems than men. Women appear to be less likely than men to manifest certain risk factors for alcohol use and problems and are more likely to have certain protective factors against these problems: women perceive greater social sanctions for drinking; women are less likely to have characteristics associated with excessive drinking including aggressiveness, drinking to reduce distress, behavioral undercontrol, sensation seeking and antisociality; and women are more likely to have desirable feminine traits (e.g., nurturance) protective against excessive drinking. In addition, consequences of heavy alcohol use, or alcohol use disorders, appear to be more negative for women than men, at least in some domains: women suffer alcohol related physical illnesses at lower levels of exposure to alcohol than men, and some studies suggest women suffer more cognitive and motor impairment due to alcohol than men; women may be more likely than men to suffer physical harm and sexual assault when they are using alcohol; heavy alcohol use in women is associated with a range of reproductive problems. Implications of these findings for future research and public health education campaigns are discussed. PMID- 15533282 TI - A multidimensional meta-analysis of psychotherapy and pharmacotherapy for obsessive-compulsive disorder. AB - A number of qualitative and meta-analytic reviews point to the efficacy of psychotherapeutic and pharmacological interventions for obsessive-compulsive disorder (OCD). In this article, we report a multidimensional meta-analysis of psychological and pharmacological treatment studies for OCD published between 1980 and 2001, examining a range of variables not previously meta-analyzed, including exclusion rates and exclusion criteria, percent of patients improved or recovered post-treatment, mean post-treatment symptomatology, and long-term outcome. These additional metrics provide a more nuanced view of the strengths and limitations of the existing data and their implications for clinical practice. Behavioral and cognitive-behavioral therapy, and a range of pharmacological interventions, lead to substantial improvement for the average patient, with individual psychotherapies and clomipramine and other Serotonin reuptake inhibitors faring best across multiple metrics. However, OCD symptoms persist at moderate levels even following adequate treatment course, and no replicable data are available on maintenance of gains for either form of treatment at 1 year or beyond. Future research should track recruitment and exclusion of study participants, include more comorbid patients, and focus on longer-term follow-up using multiple indices of outcome. More research on combined pharmacological and psychotherapeutic interventions is also indicated. PMID- 15533283 TI - The roles of depression and anxiety in the understanding and treatment of Obstructive Sleep Apnea Syndrome. AB - Obstructive Sleep Apnea Syndrome (OSAS) is a debilitating condition stemming from disruption to the respiratory system during sleep. At present, the nature of the relationship between OSAS and mood, specifically depression and anxiety, is still unclear. The purpose of this paper is to shed some light on this relationship. PsycINFO was used to locate relevant papers on this topic. This literature search formed the basis of our investigation. Results showed that the anxiety and depression methodology is weak. It is now clear that there is an urgent need to better understand the roles of anxiety and depression in OSAS. For example, the research literature suggests that depression and anxiety covary with OSAS. However, because of methodological issues, such as difficulties involved in diagnosis and the use of inappropriate instruments, this conclusion remains tenuous. Future directions are discussed. PMID- 15533284 TI - Plasma bioactive LH and testosterone profiles in male New Zealand rabbits experimentally infected with Schistosoma mansoni. AB - The effects of Schistosoma mansoni (S. mansoni) infection on plasma levels of bioactive luteinising hormone (LH) and testosterone in the New Zealand rabbit model were studied. S. mansoni infection significantly decreased the pulse frequency (P < 0.05), amplitude (P < 0.05), area under LH curve (P < 0.05) and mean plasma LH concentrations (P < 0.05) on days 42 and 70 post-infection, as compared to values for day 14 pre-infection. Areas under the response curves for plasma testosterone levels decreased significantly (P < 0.05) on days 42 and 70 post-infection in infected animals compared to day 14 pre-infection. In the praziquantel-treated group, the levels of LH and testosterone remained unchanged throughout the experimental period. The pulsatile secretion of LH was completely inhibited in S. mansoni-infected animals 70 days post-infection. These results suggest that the effects on reproductive gonadal hormones caused by S. mansoni in the rabbit model may partly be induced by alteration in pituitary synthesis or release of LH. PMID- 15533285 TI - Prevalence of intestinal parasitic infections and their relation with socio economic factors and hygienic habits in Tehran primary school students. AB - School age children carry the heaviest burden of morbidity due to intestinal parasitic infections. Our objective was to determine the prevalence of these infections in primary school children living in Tehran and their association with socio-economic factors and hygienic habits. In September 1998, a total of 19,213 subjects were invited to participate. Data on health and socio-economic status and health-related behaviours, collected via questionnaires, physical examinations and stool sample analyses, were available for 19,209 persons, with a participation rate greater than 99.99%. All participants were subjected to three methods of microscopic examinations on the stool sample and an adhesive cellophane tape slide evaluation. The prevalence rate of intestinal parasitic infection among the students was 18.4%. Coinfection with two or three parasites was seen in 2%. With increase in educational level of parents (especially mothers), the infection rate of children was decreased. Girls showed a significantly higher positive rate than boys. Using piped water, correct method of washing vegetables and increase in the economic score of the family, resulted in a decrease in the infection rate. According to the results, low level of education and consequently poor socio-economic and hygienic condition of families appear to be powerful determinants of infection. PMID- 15533286 TI - Seroepidemiology of neutralizing antibodies to Japanese encephalitis virus in Singapore: continued transmission despite abolishment of pig farming? AB - Japanese encephalitis virus (JEV) transmission in Singapore appeared to have ceased after pig farming in Singapore was phased out from the early 1980s. However, the recent detection of neutralizing antibodies to JEV in a population of wild boars in an offshore island, as well as the notification of two human cases of JE in Singapore in 2001, prompted us to reconsider the presence and hence the public health threat of JEV in Singapore. We report here a serological study of animals, birds and humans for neutralizing antibodies to JEV. The results indicate that JEV may still be actively transmitted in the peripheral part of the Singapore island and that regular serological surveys of farm animals and birds, such as chickens, may be useful to further elucidate the activity of JEV in Singapore. PMID- 15533287 TI - Susceptibility of TNF-alpha-deficient mice to Trypanosoma congolense is not due to a defective antibody response. AB - C57BL/6 mice deficient in one or two copies of the gene for tumor necrosis factor alpha (TNF-alpha) were more susceptible to Trypanosoma congolense infection than their resistant, wild-type counterparts. The number of TNF-alpha genes was correlated with the capacity to control parasitaemia and with survival time. Absence of TNF-alpha resulted in a diminished capacity to form germinal centres in lymph nodes and spleen. Since germinal centres are involved in antibody production and affinity maturation, the susceptibility of the TNF-alpha-deficient mice could have been due to this secondary defect. Despite the lack of the germinal centres, the antibody responses to internal and exposed trypanosome antigens and to non-trypanosome antigens were not significantly different. Also the relative avidities measured in infected sera did not significantly differ between the two mouse strains. These data suggest that the role of TNF-alpha in control of T. congolense was not due to its role in the development of an antibody response. PMID- 15533288 TI - Variations in helminth faecal egg counts in Kato-Katz thick smears and their implications in assessing infection status with Schistosoma mansoni. AB - Examination of stool specimens by Kato-Katz (K-K) thick smears is the standard method recommended by the WHO for field diagnosis of intestinal schistosomiasis. However, there is increasing concern that this technique has low diagnostic sensitivity. In 326 study subjects, we compared the diagnostic yield of examining one, three or five Kato-Katz thick smears prepared from one stool specimen using 41.7 mg templates. In a subset of 169 subjects who had no demonstrable Schistosoma mansoni eggs in their first three Kato-Katz thick smears, we assessed the comparative advantage of examining an additional three Kato-Katz thick smears from another stool specimen, taken four weeks later, to that of cumulative yield obtained by examining all five Kato-Katz thick smears derived from the first stool specimen. For all helminth infections, single Kato-Katz thick smear-based prevalence estimates were significantly lower than those obtained from triplet or quintet Kato-Katz thick smears. Prevalence of S. mansoni infection based on single, triplet and quintet Kato-Katz thick smears from one stool specimen were 31.3%, 45.7% and 52.1%, respectively. Prevalence estimate of S. mansoni based on quintet Kato-Katz thick smears from the first day stool specimens was not different from cumulative estimate obtained with two triplet Kato-Katz thick smears from two stool specimens, 52.1% and 52.8%, respectively. In conclusion, either examination of quintet Kato-Katz thick smears from one stool specimen using 41.7 mg template or initial triplet Kato-Katz thick smears from one stool specimen, and if these are negative, followed by examination of additional triplet Kato-Katz thick smears from subsequent day stool specimen can adequately assess individuals for infection status with S. mansoni. PMID- 15533289 TI - Occurrence of peri-parturient rise in trichostrongylid nematode egg output in Dorper ewes in a semi-arid area of Kajiado District of Kenya. AB - An investigation on the occurrence of peri-parturient rise in trichostrongylid nematode egg output in breeding ewes was carried out on a Ranch in the semi-arid area of Kajiado District in Kenya during the period May 1999-April 2000. During the study, 20 ewes randomly selected from the breeding stock and 20 un-mated female yearlings were monitored for faecal strongyle egg output every 3 weeks. A significant peri-parturient rise in faecal egg output occurred at around the time of lambing and throughout the lactation period in the mated ewes, but not in the un-mated yearlings. The occurrence of peri-parturient rise in breeding ewes towards the end of the dry season and at the onset of the short rains was enhanced by the resumption of development of hypobiotic larvae. Self-cure occurred in the un-mated yearlings in September 1999 during the dry season and in November-December 1999 during the wet season, but was less obvious in the pregnant and lactating ewes. The peri-parturient ewes contributed to higher pasture contamination at a time when highly susceptible lambs were grazing alongside. Therefore, the control of gastrointestinal nematode parasites in ewes in this area should aim at reducing the effects of this phenomenon through treatment of ewes about 2-3 weeks to lambing and during lactation. PMID- 15533290 TI - Species co-occurrence and feeding behavior in sand fly transmission of American cutaneous leishmaniasis in western Venezuela. AB - The structure of the Phlebotomine sand fly community from the Venezuelan Andes was studied using null-model tests. The analyses, at the living zones and altitudes scales, revealed C-scores larger than those expected by random, independently of the collection technique (P < 0.05). These results imply that sand fly species are non-aggregated at both scales. Random results for the variance of C-score and for the favored states hypothesis suggest that sand fly species belong to an unique guild. The latter is reinforced by the fact that anthropophilic and zoophilic species use in the same way a common resource (blood). Finally, we suggest additional approaches to study the role of the sand fly community structure on the genesis and dynamics of transmission of American cutaneous leishmaniasis. PMID- 15533291 TI - High infection rate of Wolbachia endobacteria in the sand flea Tunga penetrans from Brazil. AB - Tunga penetrans is an ectoparasite causing considerable morbidity in endemic communities. Recently, endobacteria of the genus Wolbachia were identified also in T. penetrans. Since Wolbachia were suggested as targets for intervention of insect pests and human filariasis, sand fleas were collected from infested humans, dogs and rats in a hyperendemic area in northeastern Brazil, and screened for Wolbachia infections. Twenty-one adult fleas and four batches of flea eggs were examined by PCR using primers targeting the 16S rDNA, the DNA coding for FtsZ cell-cycle protein or a Wolbachia surface protein (WSP-1). Wolbachia were detected in all examined samples from eggs, free-living male and female fleas and from neosomic female fleas. No Wolbachia DNA was detected in two samples containing flea faeces. In addition, Wolbachia were labelled by immunohistology in the ovaries of 37 female fleas using antisera raised against WSP-1 of Wolbachia the filarial parasite Dirofilaria immitis. In the vicinity of the embedded fleas containing the Wolbachia, infiltrations of neutrophils and macrophages were observed. This study showed that Wolbachia endobacteria are abundant in T. penetrans and that all examined fleas were infected by these endobacteria. Our findings may have important implications for the future development of control strategies for human tungiasis. PMID- 15533292 TI - Echinococcus granulosus cattle strain identification in an autochthonous case of cystic echinococcosis in central Mexico. AB - Echinococcosis is a frequent hepatic parasitic disease in several countries but it is practically absent in Mexico. A cattle strain of Echinococcus granulosus was identified by RAPD, PCR-RFLP and mitochondrial CO1 gene analysis in an autochthonous case. The parasite was obtained after a laparoscopic excision of a liver cyst from a patient that was symptomatic for 6 years but mis-diagnosed before hospitalization. PMID- 15533293 TI - In vitro activity of two phenyl-carbamate derivatives, singly and in combination with albendazole against albendazole-resistant Giardia intestinalis. AB - Giardia intestinalis is one of the most prevalent parasites in adults and children in Mexico. Benzimidazoles have been proposed as a therapeutic alternative in the treatment of giardiasis. However, high-dose related toxicity and the development of resistance have emerged in clinical trials using this therapy. In the search of alternative drugs, we found that benzimidazole resistant strains of fungi have shown increased sensitivity to phenyl-carbamates, hence, we developed several substituted phenyl-carbamates, two of which were tested against the protozoan parasite G. intestinalis in susceptible and albendazole-resistant Giardia strains. 4-R-ethyl-phenyl-carbamates IRE-6A and IRE 7B demonstrated antigiardial, albeit modest, activity when compared with albendazole, against susceptible and albendazole-induced resistant Giardia. However, when albendazole 0.38 microg/mL (MIC(50)) was combined with each IRE compound, a significant antigiardial synergism (fractional inhibitory concentration index (FICI < 0.5)) was obtained not only with sensitive cultures but also with resistant Giardia parasites. The results described here suggest a potential role for a combined therapy with phenyl-carbamates and sub-doses of benzimidazoles in the treatment of giardiasis. PMID- 15533294 TI - Mathematical modelling of American cutaneous leishmaniasis: incidental hosts and threshold conditions for infection persistence. AB - We present a model for the dynamics of transmission of American Cutaneous Leishmaniasis (ACL) that includes a population of incidental hosts for parasites (that is, they act only as sinks of infection), along with species that are reservoir hosts (that is, both source and sink of infection). Although, there is evidence that suggests the existence of incidental hosts for Leishmania parasites, there are no mathematical models developed to account for this. Based on this model we obtained expressions that allow computing the threshold conditions for the persistence of the infection using three different approaches. The three expressions agree in the parameters involved in the computation of the threshold, and also in the parameters that are excluded from it, which are those related to the transmission in incidental hosts. We also suggest alternatives for future development in the modelling of the dynamics of transmission of ACL, according to the needs of a more realistic, precise, or general character of the model. PMID- 15533295 TI - Detection of specific cysticercus antigen in the urine for diagnosis of neurocysticercosis. AB - Neurocysticercosis (NCC) is an important zoonotic infection, and a significant cause of morbidity and mortality in India. Immunodiagnosis plays an important role in the early, rapid, and specific diagnosis of the condition. The aim of the present study was to detect specific cysticercal antigen excreted in the urine for the diagnosis of NCC. The objective was to develop and evaluate staphylococcal Co-agglutination (Co-A) test for the detection of urinary cysticercal antigen. In the present study, urine samples were collected from 9 cases of clinically suspected NCC, 8 cases of CT/MRI proven cases of NCC, 11 of non-cysticercal CNS infection controls, and 25 healthy control subjects. Polyclonal antisera raised in rabbits against porcine complete cysticercus homogenate antigen, was used in the Co-A test to detect cysticercal antigen. The Co-A detected excreted cysticercal antigen in the urine of five of nine (55.5%) clinically diagnosed NCC and in five of eight (62.5%), CT/MRI proven cases of NCC. A false positive reaction was observed with 1 of 11 (9%) of control urine specimens from non-cysticercal CNS infection controls. No false positivity was demonstrated with 25 urine samples collected from healthy controls. Results of the present study shows that the Co-A test can be employed as a moderately sensitive and specific test for detection of urinary antigen in the diagnosis of NCC in poorly equipped laboratories. PMID- 15533296 TI - In vitro screening for antiplasmodial activity of isoquinoline alkaloids from Brazilian plant species. AB - In the search for new antimalarial agents, nine Brazilian plant species were selected, from the Annonaceae (6), Menispermaceae (2) and Siparunaceae (1) families naturally occurring at the cerrado and Atlantic rainforest regions, in order to investigate their in vitro antiplasmodial activity. The ethanol and the alkaloid extracts were tested against K1, chloroquine-resistant, and Palo Alto, chloroquine-sensitive, strains of Plasmodium falciparum. The majority of the alkaloid extracts were more active than the ethanol ones, with IC(50) ranging 0.3 8.2 microg/mL. The crude Guatteria australis alkaloids were the most active against K1 with an IC(50) = 0.3 microg/mL. The most promising total alkaloid fractions for further bioguided isolation are those with the IC(50) < or = 5 microg/mL: G. australis, Cissampelos ovalifolia and Duguetia lanceolata. PMID- 15533297 TI - Efficacy of Trypan: a diminazene based drug as antileishmanial agent. AB - Trypan, a diamidine based drug, was tested as an antileishmanial agent. Duplicate cultures of both Leishmania major and Leishmania donovani promastigotes in M199 medium and Trypan at various concentrations were tested. The cultures were incubated at 25 degrees C and parasites counted at 48 h interval, and the data generated was used to establish growth inhibition curves. Drug-free cultures were included to serve as control. In the in vivo study, a total of 40 BALB/c mice were divided into five groups of 8 mice each. They were infected with 2 x 10(6) promastogotes on the left footpad. Two groups were treated with 70 microg/ml of Trypan, a total of 500 microl used immediately after infection, one group by topical application and the other administered intraperitoneally. The treatments were repeated for the two other groups 10 weeks post infection, one by topical application and the other administered intraperitoneally. One group was not treated and thus served as control. Footpad sizes were measured using Vernier calliper every 2 weeks for 21 weeks. In the in vitro studies, Trypan inhibited growth of either L. major or L. donovani promastigotes in all the concentrations tested with more dramatic inhibition in high concentrations. Based on the in vivo studies, it was evident that Trypan had effect on L. major infected lesions when applied topically immediately after infection. However, there was no effect when treatment commenced after the lesions were established. The data is discussed. PMID- 15533298 TI - An improved PCR-based method for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in formalin-fixed stools. AB - Detection of infectious agents in faecal samples by polymerase chain reaction (PCR) can be limited by the presence of substances which inhibit DNA amplification. Here, an improved protocol is reported for directly isolating DNA from fresh and aged formalin-fixed stools, after concentration by formalin-ethyl acetate (FEA). The protocol was successfully applied to detect DNA of Entamoeba histolytica/Entamoeba dispar complex in stools by nested PCR, showing high specificity and low detection limit. Extended time of specimen storage in formalin had no influence on PCR yields. This PCR-based method offers technical advantages for routine detection and discrimination of invasive E. histolytica and non-invasive E. dispar. PMID- 15533299 TI - Rapid clearance of circulating Leishmania kinetoplast DNA after treatment of visceral leishmaniasis. AB - With the aim of evaluating the utility of the detection of Leishmania kDNA in peripheral blood for the cure assessment of visceral leishmaniasis (VL), a PCR based method was performed in patients with confirmed VL at three follow-up periods after specific chemotherapy with pentavalent antimonial. In 16 out of 17 (94.1%) patients with pre-treatment detectable kDNA that were clinically cured, the PCR turned negative up to 37 days after the initiation of treatment, remaining negative over 90 days after treatment. The clearance of Leishmania kDNA from peripheral blood of patients with VL hints to occur during or shortly after treatment concurring or preceding clinical recovery. PMID- 15533300 TI - Stability of giant unilamellar vesicles and large unilamellar vesicles of liquid ordered phase membranes in the presence of Triton X-100. AB - We have investigated the stability of giant unilamellar vesicles (GUVs) and large unilamellar vesicles (LUVs) of lipid membranes in the liquid-ordered phase (lo phase) against a detergent, Triton X-100. We found that in the presence of high concentrations of Triton X-100, the structure of GUVs and LUVs of dipalmitoyl-PC (DPPC)/cholesterol (chol) and sphingomyelin (SM)/chol membranes in the lo phase was stable and no leakage of fluorescent probes from the vesicles occurred. We also found that ether-linked dihexadecylphosphatidylcholine (DHPC) membranes containing more than 20 mol% cholesterol were in the lo phase, and that DHPC/chol GUV and DHPC/chol-LUV in the lo phase were stable and no leakage of internal contents occurred in the presence of Triton X-100. In contrast, octylglucoside solution could easily break these GUVs and LUVs of the lo phase membranes and induced internal contents leakage. These data indicate that GUVs and LUVs of the lo phase membranes are very valuable for practical use. PMID- 15533301 TI - Expression of K+-Cl- cotransporters in the alpha-cells of rat endocrine pancreas. AB - The expression of K+-Cl- cotransporters (KCC) was examined in pancreatic islet cells. mRNA for KCC1, KCC3a, KCC3b and KCC4 were identified by RT-PCR in islets isolated from rat pancreas. In immunocytochemical studies, an antibody specific for KCC1 and KCC4 revealed the expression of KCC protein in alpha-cells, but not pancreatic beta-cells nor delta-cells. A second antibody which does not discriminate among KCC isoforms identified KCC expression in both alpha-cell and beta-cells. Exposure of isolated alpha-cells to hypotonic solutions caused cell swelling was followed by a regulatory volume decrease (RVD). The RVD was blocked by 10 microM [dihydroindenyl-oxy] alkanoic acid (DIOA; a KCC inhibitor). DIOA was without effect on the RVD in beta-cells. NEM (0.2 mM), a KCC activator, caused a significant decrease of alpha-cell volume, which was completely inhibited by DIOA. By contrast, NEM had no effects on beta-cell volume. In conclusion, KCCs are expressed in pancreatic alpha-cells and beta-cells. However, they make a significant contribution to volume homeostasis only in alpha-cells. PMID- 15533302 TI - Identification of TrkA on living PC12 cells by atomic force microscopy. AB - In neural cells, nerve growth factor (NGF) initiates its survival signal through the binding to its cell surface receptor tyrosine kinase A (TrkA). Understanding the pattern of TrkA distribution and association in living cells can provide a fingerprint for the diagnostic comparison with alterations underlying ligand receptor dysfunction seen in various neurological diseases. In this study, we use the NGF-TrkA-specific interaction as a probe to identify TrkA on living PC12 cell by atomic force microscopy (AFM). An NGF-modified AFM tip was used to perform force volume (FV) imaging, generating a 2D force map to illustrate the distribution and association of TrkA on PC12 cell membrane. It is found that TrkA is highly aggregated at local regions of the cell. This unique protein association may be required to promote its function as a receptor of NGF. The methodology that we developed in this study can be adapted by other systems, thus providing a general tool for investigating protein association in its natural environment. PMID- 15533303 TI - Influence of the lipid composition on the kinetics of concerted insertion and folding of melittin in bilayers. AB - We have examined the kinetics of the adsorption of melittin, a secondary amphipathic peptide extracted from bee venom, on lipid membranes using three independent and complementary approaches. We probed (i) the change in the polarity of the 19Trp of the peptide upon binding, (ii) the insertion of this residue in the apolar core of the membrane, measuring the 19Trp-fluorescence quenching by bromine atoms attached on lipid acyl chains, and (iii) the folding of the peptide, by circular dichroism (CD). We report a tight coupling of the insertion of the peptide with its folding as an alpha-helix. For all the investigated membrane systems (cholesterol-containing, phosphoglycerol containing, and pure phosphocholine bilayers), the decrease in the polarity of 19Trp was found to be significantly faster than the increase in the helical content of melittin. Therefore, from a kinetics point of view, the formation of the alpha-helix is a consequence of the insertion of melittin. The rate of melittin folding was found to be influenced by the lipid composition of the bilayer and we propose that this was achieved by the modulation of the kinetics of insertion. The study reports a clear example of the coupling existing between protein penetration and folding, an interconnection that must be considered in the general scheme of membrane protein folding. PMID- 15533304 TI - Oligomerization triggers binding of a Bacillus thuringiensis Cry1Ab pore-forming toxin to aminopeptidase N receptor leading to insertion into membrane microdomains. AB - Bacillus thuringiensis Cry1A toxins, in contrast to other pore-forming toxins, bind two putative receptor molecules, aminopeptidase N (APN) and cadherin-like proteins. Here we show that Cry1Ab toxin binding to these two receptors depends on the toxins' oligomeric structure. Toxin monomeric structure binds to Bt-R1, a cadherin-like protein, that induces proteolytic processing and oligomerization of the toxin (Gomez, I., Sanchez, J., Miranda, R., Bravo A., Soberon, M., FEBS Lett. (2002) 513, 242-246), while the oligomeric structure binds APN, which drives the toxin into the detergent-resistant membrane (DRM) microdomains causing pore formation. Cleavage of APN by phospholipase C prevented the location of Cry1Ab oligomer and Bt-R1 in the DRM microdomains and also attenuates toxin insertion into membranes despite the presence of Bt-R1. Immunoprecipitation experiments demonstrated that initial Cry1Ab toxin binding to Bt-R1 is followed by binding to APN. Also, immunoprecipitation of Cry1Ab toxin-binding proteins using pure oligomeric or monomeric structures showed that APN was more efficiently detected in samples immunoprecipitated with the oligomeric structure, while Bt-R1 was preferentially detected in samples immunoprecipitated with the monomeric Cry1Ab. These data agrees with the 200-fold higher apparent affinity of the oligomer than that of the monomer to an APN enriched protein extract. Our data suggest that the two receptors interact sequentially with different structural species of the toxin leading to its efficient membrane insertion. PMID- 15533305 TI - Identification of a cation-specific channel (TipA) in the cell wall of the gram positive mycolata Tsukamurella inchonensis: the gene of the channel-forming protein is identical to mspA of Mycobacterium smegmatis and mppA of Mycobacterium phlei. AB - Detergent extracts of whole cells of the Gram-positive bacterium Tsukamurella inchonensis ATCC 700082, which belongs to the mycolata, were studied for the presence of ion-permeable channels using lipid bilayer experiments. One channel with a conductance of about 4.5 nS in 1 M KCl was identified in the extracts. The channel-forming protein was purified to homogeneity by preparative SDS-PAGE. The protein responsible for channel-forming activity had an apparent molecular mass of about 33 kDa as judged by SDS-PAGE. Interestingly, the protein showed cross reactivity with polyclonal antibodies raised against a polypeptide derived from MspA of Mycobacterium smegmatis similarly as the cell wall channel of Mycobacterium phlei. Primers derived from mspA were used to clone and sequence the gene of the cell wall channels of T. inchonensis (named tipA for T. inchonensis porin A) and M. phlei (named mppA for M. phlei porin A). Surprisingly, both genes, tipA and mppA, were found to be identical to mspA of M. smegmatis, indicating that the genomes of T. inchonensis, M. phlei and M. smegmatis contain the same genes for the major cell wall channel. RT-PCR revealed that tipA is transcribed in T. inchonensis and mppA in M. phlei. The results suggest that despite a certain distance between the three organisms, their genomes contain the same gene coding for the major cell wall channel, with a molecular mass of 22 kDa for the monomer. PMID- 15533306 TI - Components and fractions for differently bound water molecules of dipalmitoylphosphatidylcholine-water system as studied by DSC and 2H-NMR spectroscopy. AB - Differently bound water molecules of dipalmitoylphosphatidylcholine (DPPC)-H2O system were investigated with differential scanning calorimetry (DSC). According to a method previously reported by us, the ice-melting DSC curves of the DPPC-H2O samples of varying water contents were deconvoluted into multiple components, and the ice-melting enthalpies for the individual deconvoluted components were used to estimate average molar ice-melting enthalpies for freezable interlamellar and bulk waters, respectively. With these average molar ice-melting enthalpies, the numbers of differently bound water molecules of the DPPC-H2O system were calculated at varying water contents and were used to construct a water distribution diagram of this system. Furthermore, to evaluate the reliability of the present DSC deconvolution method, 2H-NMR T1 measurements of DPPC-2H2O system were carried out at 5 degrees C of the gel phase temperature, and components and fractions for differently bound water (2H2O) molecules were estimated from the analysis of nonexponential magnetization recovery curves. PMID- 15533307 TI - The interactions of the HIV gp41 fusion peptides with zwitterionic membrane mimics determined by NMR spectroscopy. AB - The wild-type (wt) N-terminal 23-residue fusion peptide (FP) of the human immunodeficiency virus (HIV) fusion protein gp41 and its V2E mutant have been studied by nuclear magnetic resonance (NMR) spectroscopy in dodecylphosphocholine (DPC) micelles as membrane mimics. A number of NMR techniques have been used. Pulsed field-gradient diffusion measurements in DPC and in 4:1 DPC/sodium dodecylsulfate mixed micelles showed that there is no major difference between the partition coefficients of the fusogenic wt peptide and the V2E mutant in these micelles, indicating that there is no correlation between the activity of the fusion peptides and their membrane affinities. The nuclear Overhauser enhancement (NOE) patterns and the chemical shift index for these two peptides indicated that both FP are in an alpha helical conformation between the Ile4 to Leu12 or to Ala15 region. Simulated annealing showed that the helical region extends from Ile4 to Met19. The two FPs share similar conformational characteristics, indicating that the conformation of the FP is not an important factor determining its activity. The spin-label studies, utilizing spin labels 5- and 16-doxystearic acids in the DPC micelles, provided clear indication that the wt FP inserts its N-terminus into the micelles while the V2E mutant does not insert into the micelles. The conclusion from the spin-label results is corroborated by deuterium amide proton exchange experiments. The correlation between the oblique insertion of the FP and its fusogenic activity is in excellent agreement with results from our molecular dynamics simulation and from other previous studies. PMID- 15533308 TI - Lactadherin binds selectively to membranes containing phosphatidyl-L-serine and increased curvature. AB - Lactadherin, a milk protein, contains discoidin-type lectin domains with homology to the phosphatidylserine-binding domains of blood coagulation factor VIII and factor V. We have found that lactadherin functions, in vitro, as a potent anticoagulant by competing with blood coagulation proteins for phospholipid binding sites [J. Shi and G.E. Gilbert, Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid binding sites, Blood 101 (2003) 2628-2636]. We wished to characterize the membrane-binding properties that correlate to the anticoagulant capacity. We labeled bovine lactadherin with fluorescein and evaluated binding to membranes of composition phosphatidylserine/phosphatidylethanolamine/phosphatidylcholine, 4:20:76 supported by 2 mum diameter glass microspheres. Lactadherin bound saturably with an apparent KD of 3.3+/-0.4 nM in a Ca++ -independent manner. The number of lactadherin binding sites increased proportionally to the phosphatidylserine content over a range 0-2% and less rapidly for higher phosphatidylserine content. Inclusion of phosphatidylethanolamine in phospholipid vesicles did not enhance the apparent affinity or number of lactadherin binding sites. The number of sites was at least 4-fold higher on small unilamellar vesicles than on large unilamellar vesicles, indicating that lactadherin binding is enhanced by membrane curvature. Lactadherin bound to membranes with synthetic dioleoyl phosphatidyl-L serine but not dioleoyl phosphatidyl-D-serine indicating stereoselective recognition of phosphatidyl-L-serine. We conclude that lactadherin resembles factor VIII and V with stereoselective preference for phosphatidyl-L-serine and preference for highly curved membranes. PMID- 15533309 TI - Membrane-permeabilizing activities of amphidinol 3, polyene-polyhydroxy antifungal from a marine dinoflagellate. AB - Amphidinols, which are polyene-polyhydroxy metabolites produced by the marine dinoflagellate Amphidinium klebsii, possess potent antifungal and hemolytic activities. The membrane permeabilizing actions of amphidinol 3, the most potent homologue, were compared with those of polyene antibiotics, amphotericin B (AmB) and filipin, in hemolytic tests, 23Na nuclear magnetic resonance (NMR)-based membrane permeabilizing assays, and UV spectroscopy for liposome-bound forms. In Na+ flux experiments using large unilamellar vesicles (LUVs), ion efflux by amphidinol 3 was inhibited by cholesterol or ergosterol, which was opposed to previous results [J. Mar. Biotechnol., 5 (1997) 124]. When the effect of the agents on the size of vesicles was examined by light scattering experiments, amphidinol 3 did not significantly alter their size while filipin and synthetic detergent Triton X-100 did. The observations implied that the activity of amphidinol 3 was mainly due to formation of large pores/lesions in liposomes rather than detergent-like disruption of membrane. The pore/lesion size was estimated to be 2.0-2.9 nm in diameter on the basis of osmotic protection experiments using blood cells. The UV spectra in liposomes, which revealed the close interaction of polyene moieties in a lipid bilayer, further implied that the membrane activity of amphidinol 3 is caused by the molecular assemblage formed in biomembrane. These results disclose that amphidinol 3 is one of few non ionic compounds that possess potent membrane permeabilizing activity with non detergent mechanism. PMID- 15533310 TI - Anesthetized Long Evans rats show similar protein expression and long-term potentiation as Fischer 344 rats but reduced short-term potentiation in motor cortex. AB - A number of studies describe strain-related differences in the motor behavior of rats. Inbred albino F344 rats are found to be impaired in procedural spatial learning, skilled reaching, and over ground locomotion in relation to pigmented out bred Long Evans (LE) rats. These deficits could be related to the functional differences in the motor cortex of the two strains, and the objective of the present study was to examine this hypothesis. Synaptic transmission was examined in the two rat strains, using long-term potentiation (LTP) and short-term potentiation (STP), two electrophysiological measures of neural function and learning. Field potentials were evoked in the motor cortex of anesthetized Long Evans and Fischer 344 (F344) rats in response to contralateral white matter stimulation. The main findings indicated that (1) baseline-evoked responses in the two strains was similar, indicating similar basal levels of synaptic strength, (2) LTP was induced in both strains of rats, suggesting similar synaptic efficacy in the two strains of rats, and (3) STP was enhanced in the Fischer 344 rats, suggesting differences in synaptic function. Protein expression also revealed that the two strains did not differ with respect to structural or synaptic protein expression. Thus, the two strains exhibit motor skill differences despite a great degree of physiological similarity in motor cortex. The results are discussed in relation to the greater utility of using the Long Evans rat for examining the neural basis of plasticity and models of disease, especially if motor tasks are evaluated. PMID- 15533311 TI - Brain-derived neurotrophic factor mRNA and protein in the piglet brainstem and effects of Intermittent Hypercapnic Hypoxia. AB - Brain-derived neurotrophic factor (BDNF) is a neurotrophin essential for the development of normal respiratory rhythm and ventilatory control. Chronic exposure to Intermittent Hypercapnic Hypoxia (IHH) has been shown to alter ventilatory responses of piglets. This study investigated changes in BDNF distribution and expression in seven nuclei of the caudal medulla, from piglets exposed to IHH for 1, 2, 3, or 4 days before death, using non-radioactive in situ hybridisation (for mRNA) and immunohistochemistry (for protein). Compared to controls, BDNF mRNA was markedly increased across the entire medulla of the brainstem, after all durations of IHH (1-4 days). In contrast, BDNF protein expression increased after 1 day of exposure to IHH (p=0.003), but, thereafter, was not different to controls. Amongst individual nuclei, neurons of the dorsal motor nucleus of the vagus (DMNV) showed increased BDNF mRNA (p<0.01), but decreased protein expression (p=0.05) after all durations of IHH. In the ION, both mRNA and protein for BDNF were significantly increased after 1 day IHH (p<0.01 and p=0.001, respectively), but these increases were not sustained. This study is the first to investigate changes in BDNF expression in response to environmental challenges during postnatal development in the brainstem. Implications of the wide distribution of BDNF in the piglet caudal medulla and increased expression after IHH exposure are discussed, with particular reference to roles for BDNF-dependent neurons at this stage of development. PMID- 15533312 TI - Fluoro-Jade, but not Fluoro-Jade B, stains non-degenerating cells in brain and retina of embryonic and neonatal rats. AB - Fluoro-Jade (FJ) and Fluoro-Jade B (FJB) are fluorescein derivatives currently used to stain brain cells under degeneration. In this study, we investigated the FJ staining of nondegenerating cells in embryonic and neonatal rat brain and retina. In embryonic rat brain (embryonic day 15; E15), very intense staining of cells was observed. The number of FJ-stained cells and the intensity of staining decreased with increasing in animal age, being almost absent by postnatal day 16 (P16). Only a few cells in neonatal rat brain were in the process of cell death, as verified by the TUNEL technique. The FJ-stained cells in neonatal brain were positive for the neuronal marker neuronal nuclei antigen (NeuN). In retina, FJ stained mainly cells from the ganglion cell layer at P2 and the neuroblastic layer at P2 and P6. In contrast to FJ, FJB did not stain nondegenerating cells in embryonic and neonatal rats. These results show that in addition to staining degenerating brain cells, FJ also stains nondegenerating central nervous system cells in embryonic and neonatal stages. PMID- 15533313 TI - Suppressive effects of milk-derived lactoferrin on psychological stress in adult rats. AB - Lactoferrin (LF) is known as an iron-binding glycoprotein. It has been shown that bovine LF (bLF) is transported into cerebrospinal fluid via blood although its physiological effects in the central nervous system (CNS) are still unclear. In this study, a suppressive effect of bLF on psychological distress was investigated in adult rats. Intraperitoneal injection of bLF (100 mg/kg) reduced stressful behaviors in a conditioned fear-induced freezing test and an elevated plus-maze test. Interestingly, the suppressive effect of bLF was enhanced by pretreatment with electric foot-shock (FS). This suppressive effect of bLF in the elevated plus-maze test was reversed by pretreatment with naloxone, an opioid receptor antagonist, at a dose of 1 mg/kg (ip). N(omega)-nitro-l-arginine methyl ester (l-NAME), a nitric oxide synthase (NOS) inhibitor, also blocked the suppressive effect of bLF and foot-shock. In addition, combined application of a low dose of bLF (30 mg/kg, ip) and l-arginine (30 and 100 mg/kg, ip) showed significant potentiated effects on psychological stress. These results suggest that bLF has suppressive effects on psychological distress, especially under the condition of moderate stress. Furthermore, it is suggested that bLF possibly activates an endogenous opioidergic system via nitric oxide synthase activation. PMID- 15533314 TI - Effects of acute and chronic lamotrigine treatment on basal and stimulated extracellular amino acids in the hippocampus of freely moving rats. AB - The antiepileptic drug lamotrigine (LTG) is a relatively novel anticonvulsant frequently used in polytherapy and increasingly in monotherapy. LTG is believed to act by reducing excitatory glutamate (GLU) release due to an inhibition of Na(+) channels. In the present study, we have investigated the effects of acute and chronic (up to 21 days) treatment with LTG on basal and either veratridine- or KCl-stimulated release of aspartate (ASP), GLU, taurine (TAU) and GABA in the hippocampus of freely moving rats using microdialysis. Additionally, we have measured LTG concentrations in the plasma, whole brain and extracellular fluid of rats at the same time points. LTG significantly reduced basal ASP and GLU but only at the highest dose used (20 mg/kg) and was entirely without effect on basal TAU or GABA. When either veratridine or 100 mM KCl were added to the infusion medium amino acid release was evoked although the extent of this varied from one amino acid to another. LTG (10 mg/kg) reduced veratridine-evoked release of all four amino acids studied, although this was most marked in the case of GLU. LTG had no effect on KCl-stimulated amino acid release. When given for up to 21 days (2 x 5 mg/kg/day), LTG had no effect on basal amino acid levels. In contrast, LTG demonstrated over the time period studied an increasingly inhibitory effect on veratridine-evoked amino acid release. This effect of the drug was proportionally much greater in the case of GLU than for the other three amino acids studied. Measurement of plasma, whole brain tissue and extracellular LTG showed that in each of these compartments, it had reached an apparent steady state within 4 days of commencement of treatment and appeared to mirror the neurochemical changes measured. Our estimate of plasma LTG indicates that during chronic study, this was well within the therapeutic range, suggesting that the current neurochemical observations are clinically relevant. PMID- 15533315 TI - Lithium inhibits the modulatory effects of morphine on susceptibility to pentylenetetrazole-induced clonic seizure in mice: involvement of a nitric oxide pathway. AB - Lithium has been reported to inhibit opioid-induced properties. The present study examined the effect of acute and chronic administration of lithium chloride (LiCl) on morphine's biphasic modulation of susceptibility to pentylenetetrazole (PTZ)-induced clonic seizure in mice. We also examined the possible involvement of nitric oxide (NO) pathway in lithium effect. Both acute (0.1 and 1 mg/kg) and chronic (same doses, 21 consecutive days) administration of LiCl completely inhibited the anticonvulsant and proconvulsant effects of morphine (at doses 1 and 30 mg/kg, respectively). A very low and per se noneffective dose of LiCl (0.05 mg/kg) significantly inhibited both phases of morphine effect when administered concomitant with a noneffective low dose of naloxone (0.1 mg/kg). The NO synthase inhibitor N(G)-nitro-l-arginine methyl ester (L-NAME) at a per se noneffective dose of 0.3 mg/kg potentiated the inhibitory effects of low doses of LiCl (0.01 and 0.05 mg/kg) on both phases of morphine effect. l-arginine, a NO synthase substrate, at a per se noneffective dose of 30 mg/kg reversed the inhibitory effects of lithium (1 mg/kg). Lithium is capable of antagonizing both modulatory effects of morphine on seizure susceptibility even at relatively low doses. These inhibitory effects of lithium may also involve NO synthesis. PMID- 15533316 TI - Intravitreal transplants of Schwann cells and fibroblasts promote the survival of axotomized retinal ganglion cells in rats. AB - Schwann cells (SCs) are considered one of the major cellular components to maintain the integrity of the peripheral nervous system (PNS) neurons after injury. Intravitreal transplant of peripheral nerves or Schwann cells has been shown to enhance the regenerative ability of retinal ganglion cells (RGCs). In the present study, we compared the effects of intravitreal transplants of Schwann cells and fibroblasts, two major components of peripheral nerves, on the survival of retinal ganglion cells in adult rats after optic nerve (ON) transection. Purified Schwann cells and fibroblasts from neonatal sciatic nerves were injected into the vitreous body of adult rats. Three days after the injection, the optic nerves were transected intraorbitally. After 1 week or 1 month, surviving retinal ganglion cells were retrogradely labelled with Fluoro-Gold (FG) and the number of surviving retinal ganglion cells was counted. The retinas were further processed for 200-kDa neurofilament RT-97 immunohistochemistry. It was found that intravitreally injected- Schwann cells and -fibroblasts delayed the death of axotomized retinal ganglion cells for 1 week. In addition, in the animal group with 1 month survival time after optic nerve transection, those received a larger number of Schwann cells had more surviving retinal ganglion cells and more profusely ramified axonal processes near the optic disc. These findings reveal that both Schwann cells and fibroblasts isolated from the peripheral nerve can promote retinal ganglion cell survival after optic nerve transection, presumably by secreting neurotrophic factors. In addition, the data also demonstrate that Schwann cells could promote intraretinal axonal sprouting. Our findings demonstrate a remarkable glial source of neurotrophic factors with potential clinical applications, as autologous Schwann cells and fibroblasts can be feasibly obtained from peripheral nerves. PMID- 15533317 TI - The behavioral and neuroanatomical effects of IB4-saporin treatment in rat models of nociceptive and neuropathic pain. AB - One distinguishing feature of primary afferent neurons is their ability to bind the lectin IB(4). Previous work suggested that neurons in the inner part of lamina II (IIi), onto which IB(4)-positive sensory neurons project, facilitate nociceptive transmission following tissue or nerve injury. Using an IB(4)-saporin conjugate (IB(4)-SAP), we examined the contribution of IB(4)-positive neurons to nociceptive processing in rats with and without nerve injury. Intrasciatic injection of IB(4)-SAP (5 mug/5 mul) significantly decreased IB(4)-labeling and immunoreactive P(2)X(3) in the spinal cord and delayed the behavioral and neuroanatomical consequences of L5 spinal nerve ligation (SNL) injury. In the absence of injury, thermal and mechanical nociceptive thresholds increased 2 weeks post-treatment only in IB(4)-SAP-treated, but not control (saline or saporin only), rats. Acute NGF-induced hyperalgesia was also attenuated following IB(4)-SAP treatment. In the SNL model, mechanical allodynia failed to develop 1 and 2 weeks post-injury, but was fully established by 4 weeks. Moreover, neuropeptide Y immunoreactivity (NPY-ir), which increases in the spinal cord after nerve injury, was unchanged in IB(4)-SAP-treated animals whereas immunoreactive PKCgamma decreased 2, but not 4, weeks post-injury. Quantitative RT-PCR revealed a reduction in P(2)X(3) mRNA in L4 DRG of IB(4)-SAP-treated animals, but no change in TrkA expression. Our results suggest that IB(4) positive neurons in L4 are required for the full expression of NGF-induced hyperalgesia and participate in the behavioral and anatomical consequences that follow injury to the L5 spinal nerve. PMID- 15533318 TI - Neural substrates for sexual and thermoregulatory behavior in the male leopard gecko, Eublepharis macularius. AB - The preoptic area-anterior hypothalamus (POAH) continuum is critical for the integration of environmental, physiological, and behavioral cues associated with reproduction in vertebrates. In the present study, radiofrequency lesions in the POAH abolished sexual behavior in the leopard gecko (Eublepharis macularius). Furthermore, results suggest a differential effect of POAH lesions on those behaviors regarded as appetitive (tail vibration and grip) and those regarded as consummatory (mounting and copulation), with consummatory behaviors being affected to a greater extent. E. macularius is an ectothermic vertebrate that modulates body temperature behaviorally relative to ambient temperature. In vertebrates, the POAH is also an important integrator of thermoregulation. Thus, the present study investigated whether lesions that disrupt reproductive behavior also disrupt body temperature regulation. While virtually all males displayed diurnal rhythms in thermoregulatory behavior prior to surgery, this pattern was abolished in a small proportion of animals bearing POAH lesions. Lesions that abolished thermoregulatory rhythms involved the suprachiasmatic nucleus (SCN), whereas lesions confined to the POAH, while dramatically influencing sexual behavior, did not affect thermoregulatory rhythms or temperature set point. Together, these findings identify the POAH as an important neural locus regulating sexual behavior but not thermoregulation and suggest that the SCN acts as a pacemaker controlling daily behavioral temperature regulation in this species. PMID- 15533319 TI - The pre-ischaemic neuroprotective effect of a novel polyamine antagonist, N1 dansyl-spermine in a permanent focal cerebral ischaemia model in mice. AB - The polyamine sites on the NMDA receptor complex offer a therapeutic target for focal ischaemia, potentially devoid of most side effects associated with NMDA antagonists. In this study, we investigated the effect of a novel polyamine antagonist, N(1)-dansyl-spermine (0.5-10 mg kg(-1)) in a permanent focal cerebral ischaemia model in mice, and compared its effect to that of MK-801 (0.3-3 mg kg( 1)) following administration 30 min prior to ischaemia. A battery of histological and behavioural tests was employed following permanent middle cerebral artery occlusion to assess any neuroprotective effect. Following middle cerebral artery occlusion, N(1)-dansyl-spermine (1-5 mg kg(-1)) and MK-801 (1 or 3 mg kg(-1)) caused a comparable and significant reduction in the percentage hemisphere lesion volume. Similarly, both drugs significantly reduced oedema and neurological deficit score to a similar extent. Locomotor activity in MCAO mice was not significantly improved by MK-801 or N(1)-dansyl-spermine, although N(1)-dansyl spermine induced a trend towards significant improvement. Significant improvement in rotarod performance was observed at neuroprotective doses with both drugs. Upon comparison of the profile of effects, N(1)-dansyl-spermine at least matched the effectiveness of MK-801 as a neuroprotective agent in this model. In addition, in sham-operated control mice, N(1)-dansyl-spermine was well tolerated, in contrast to the pronounced adverse effects of MK-801 on locomotor activity and rotarod performance. In conclusion, this study has shown that N(1)-dansyl spermine is as effective a neuroprotective drug as MK-801 in this model. Moreover, in contrast to MK-801, N(1)-dansyl-spermine could be a promising therapeutic candidate for stroke as it is well tolerated at neuroprotective doses in sham-operated animals. PMID- 15533320 TI - Dose-independent antinociceptive interaction of endogenous ligands at the spinal level. AB - Adenosine, agmatine and kynurenic acid are endogenous ligands acting on different (e.g. adenosine, NMDA, alpha(2)-adrenergic and imidazoline) receptors with a potential role in nociception at the spinal level. Their antinociceptive effects have already been investigated as monotherapy, but only a few studies have reported on their effects on the potency of other drugs. The purpose of the present study was carried out to analyse their interactions during continuous intrathecal co-administration in a carrageenan-induced thermal hyperalgesia model in rats. A paw withdrawal test was used for nociceptive testing. The intrathecal infusion (60 min) of these three drugs was administered alone or in combinations (kynurenic acid+adenosine or agmatine; adenosine+agmatine), which was followed by an additional 60-min observation period. Kynurenic acid alone was ineffective, while adenosine and agmatine alone caused a slight increase in pain threshold. However, independently of the applied doses all of the combinations significantly (p<0.05) increased the paw withdrawal latencies on the inflamed side during and after the infusion, but were almost ineffective on the normal side. The adenosine+kynurenic acid combination was the most effective: namely, that it relieved thermal hyperalgesia in all the applied dose combinations. Treatment with the kynurenic acid-containing combinations also caused dose-dependent side effects (motor impairment and excitation), despite the fact that monotherapy with kynurenic acid in the applied dose (0.1 microg/min) did not result in adverse effects. PMID- 15533321 TI - Role of inducible nitric oxide synthase in N-methyl-d-aspartic acid-induced strio nigral degeneration. AB - N-Methyl-d-aspartate (NMDA)-induced striatal excitotoxicity is mediated by nitric oxide (NO) but the role of inflammatory mechanisms and inducible nitric oxide synthase (iNOS) induction is not clear. Unilateral intrastriatal administration of NMDA to rats resulted in the loss of intrinsic striatal neurones and the degeneration of NADPH-diaphorase positive interneurones within 24 h. NMDA administration caused activation of glial fibrillary acidic protein positive astroglial cells and MAC-1 ir microglia. Marked iNOS immunoreactivity was expressed within both astroglial and microglial cells and there was marked cellular labelling for 3-nitrotyrosine (3-NT). One month following the NMDA lesion, administration of (+)-amphetamine (AMPH) produced a circling response in rats. Pre-treatment of rats with the iNOS inhibitor aminoguanidine (AG) decreased the extent of NMDA-induced striatal cell loss at 24 h and reduced 3-NT expression but was without effect on glial cell activation. AG pre-treatment also prevented the onset of rotation to AMPH at 30 days following NMDA lesioning. NMDA administration unexpectedly caused a loss of tyrosine hydroxylase immunoreactive (TH-ir) fibres in the striatum at 24 h and at 30 days the number of TH-ir cells were decreased in the substantia nigra. The loss of nigral cells was prevented by AG pre-treatment. This study demonstrates a role for iNOS induction in NO mediated NMDA excitotoxicity to rat striatum and suggests that inflammatory mechanisms play a key role in this process. PMID- 15533322 TI - Morphological differentiation of bone marrow stromal cells into neuron-like cells after co-culture with hippocampal slice. AB - Adult green mice marrow stromal cells were co-cultured with hippocampal slices. Differentiation to neuron-like or non-neuron-like cells occurred exclusively inside slice boundaries starting at day 3, and then decreased gradually over 35 days. Neuron-like cells tended to form network-like connections around day 14. The use of retinoic acid greatly increased the number of differentiated cells, and the most effective concentration was 10(-6) M. NeuN immunohistochemistry was positive in 9.6+/-1.7% of morphologically differentiated neuron-like cells. Both GFAP and Iba1 immunostaining were negative. We concluded that bone marrow stromal cells can be differentiated into neurons, and direct contact with the host brain tissue is essential for this to occur. Retinoic acid significantly increases the number of differentiated cells, as has been reported with other stem cells. PMID- 15533323 TI - Direct evidence for expression of dopamine receptors in astrocytes from basal ganglia. AB - Expression of dopamine receptors (DA-Rs) in astrocytes was examined in vitro and in vivo using primary cultured astrocytes and brain slices from rat basal ganglia. Astrocytes from basal ganglia expressed DA D1-, D3-, D4- and D5 receptors and D4-mediated signal transduction in response to DA, suggesting possible involvement of astrocytes in the pharmacological action of atypical antipsychotic drugs and in DA response in some neurological diseases. PMID- 15533324 TI - Muscimol injection into the lateral hypothalamus inhibits the hypotension and bradycardia caused by somato-visceral nociception. AB - This study investigated whether the lateral hypothalamus (LH) contributes to the depressor response evoked by somato-visceral nociception. Lidocaine (2%; 0.1, 0.3 or 1.0 microl) or muscimol (0.34 nmol; 0.5 microl) was microinjected into the rostral LH of halothane-anesthetized rats bilaterally and somato-visceral nociception was induced 2 min later by injecting 5% acetic acid (0.5 ml) intraperitoneally. Lidocaine and muscimol inhibited the hypotension and bradycardia caused by somato-visceral nociception significantly without affecting cardiovascular function in normotensive animals. PMID- 15533325 TI - Impairment of sensorimotor gating in mice deficient in the cell adhesion molecule L1 or its close homologue, CHL1. AB - Mutations in the gene encoding the cell adhesion molecule L1 or its close homologue, CHL1 (close homologue of L1), cause brain dysfunction in both humans and mice. Here we report that prepulse inhibition (PPI) of the acoustic startle response is impaired in mice deficient in either L1 or CHL1. This newly identified feature may provide a basis for using these mice as models for sensorimotor gating impairment found in some neuropsychiatric disorders such as schizophrenia. PMID- 15533326 TI - A bi-directional mu-opioid-opioid connection between the nucleus of the accumbens shell and the central nucleus of the amygdala in the rat. AB - The central nucleus of the amygdala (CeA) and the nucleus of the accumbens shell (NAc) have been shown to be involved in opioid-mediated feeding behavior. The present study examined whether mu-opioid signalling between the CeA and NAc affected feeding. Male Sprague-Dawley rats were fitted with one cannula placed in the CeA and two cannulae placed in the NAc, which allowed for coadministration of the mu-opioid receptor agonist [D-Ala(2), NMe-Phe(4), Gly-ol(5)]-enkephalin (DAMGO) in one site and the opioid antagonist naltrexone (NTX) in the other site. Single injection of DAMGO (2.4 nmol) into the CeA and bilateral injections of DAMGO (2.4 nmol) into the NAc stimulated feeding (P<0.05). The DAMGO-induced increase of food intake following injection into the CeA was decreased by bilateral injection of NTX (13.2 and 26.5 nmol) into the NAc at 2- and 4-h postinjections (P<0.05). In the reverse situation, the DAMGO-induced increase of food intake following injection into the NAc was decreased by injection of NTX (13.2 and 26.5 nmol) into the CeA at 1-, 2-, and 4-h postinjections (P<0.05). These results suggest that a bi-directional mu-opioid-opioid signalling pathway exists between the CeA and the NAc, which influences feeding. PMID- 15533328 TI - Maternal serum level of the DDT metabolite DDE in relation to fetal loss in previous pregnancies. AB - Use of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) continues in about 25 countries. This use has been justified partly by the belief that it has no adverse consequences on human health. Evidence has been increasing, however, for adverse reproductive effects of DDT, but additional data are needed. Pregnant women who enrolled in the Collaborative Perinatal Project (United States, 1959 1965) were asked about their previous pregnancy history; blood samples were drawn and the serum frozen. In 1997-1999, the sera of 1717 of these women who had previous pregnancies were analyzed for 1,1-dichloro-2,2-bis(p chlorophenyl)ethylene (DDE), the major breakdown product of DDT. The odds of previous fetal loss was examined in relation to DDE level in logistic regression models. Compared with women whose DDE level was <15 microg/L, the adjusted odds ratios of fetal loss according to category of DDE were as follows: 15-29 microg/L, 1.1; 30-44 microg/L, 1.4; 45-59 microg/L, 1.6; and 60+ microg/L, 1.2. The adjusted odds ratio per 60 microg/L increase was 1.4 (95% confidence interval 1.1-1.6). The results were consistent with an adverse effect of DDE on fetal loss, but were inconclusive owing to the possibility that previous pregnancies ending in fetal loss decreased serum DDE levels less than did those carried to term. PMID- 15533330 TI - Reliability of sport fish consumption in the New York State Angler cohort study. AB - This paper examines the reliability of sport fish consumption data from the New York State Angler Cohort Study (NYSACS). NYSACS is a prospective cohort study conducted among New York State registered sportsmen and fishermen. Sport fish consumption information for New York State waters including the Great Lakes between June 1990 and June 1991 were collected through self-administered questionnaires, Spouses of male anglers were asked to provide their husbands' fish consumption during the same time period. A short telephone interview after the cohort was ascertained was also conducted among about 100 study participants for the purpose of quality control. Percentage agreement, kappa, and weighted kappa were calculated to evaluate the reliability of the interview using spousal data and reinterview data. Overall, for the total fish consumption between June 1990 and June 1991, percentage agreement, kappa, and weighted kappa between spousal data and primary response were 67.28%, 0.5087, and 0.6157, respectively. For reinterview data, weighted kappa ranged from 0.4510 to 0.5285 for season specific analysis and kappa ranged from 0.4615 to 0.7006 for fish species specific analysis. Spouses may be a good source of proxy data for fish consumption. The reliability of sport fish consumption data for the NYSACS study is acceptable, suggesting that the food frequency measuring methods employed are a viable approach to obtain retrospective sport fish consumption data from sportsmen and subsistence anglers. PMID- 15533329 TI - Birth defects risk associated with maternal sport fish consumption: potential effect modification by sex of offspring. AB - Contaminated sport fish consumption may result in exposure to various reproductive and developmental toxicants, including pesticides and other suspected endocrine disruptors. We investigated the relation between maternal sport fish meals and risk of major birth defects among infants born to members of the New York State (NYS) Angler Cohort between 1986 and 1991 (n=2237 births). Birth defects (n=125 cases) were ascertained from both newborn medical records and the NYS Congenital Malformations Registry. For sport fish meals eaten during pregnancy, the odds ratio (OR) for all major malformations combined was slightly elevated for < or =1 meal/month (OR=1.26, 95% confidence interval (CI): 0.84, 1.89) and > or =2 meals/month (OR=1.51, CI=0.74, 3.09), with no meals during pregnancy as the reference category. Higher ORs were consistently observed among male offspring compared with females. For > or =2 meals/month, the risk for males was significantly elevated (males: OR=3.01, CI: 1.2, 7.5; females: OR=0.73, CI: 0.2, 2.4). Exposure during pregnancy and effect modification by infants sex could be important considerations for future studies of birth outcomes associated with endocrine disruptors. PMID- 15533331 TI - Maternal exposure to Great Lakes sport-caught fish and dichlorodiphenyl dichloroethylene, but not polychlorinated biphenyls, is associated with reduced birth weight. AB - Fish consumption may be beneficial for a developing human fetus, but fish may also contain contaminants that could be detrimental. Great Lakes sport-caught fish (GLSCF) are contaminated with polychlorinated biphenyls (PCBs) and dichlorodiphenyl dichloroethylene (DDE), but the effects of these contaminants on birth outcome are not clear. To distinguish potential contaminant effects, we examined (1) whether the decrease over time in contaminant levels in GLSCF is paralleled by an increase in birth weight of children of GLSCF-consuming mothers and (2) the relation between maternal serum concentrations of these contaminants and birth weight. Mothers (n=511) were interviewed from 1993 to 1995, and maternal serum was collected from 1994 to 1995 (n=143). Potential confounders considered were child gender, maternal age at delivery, maternal prepregnancy body mass index, maternal cigarette and alcohol use during pregnancy, maternal education level, maternal parity, and maternal breastfeeding. Children born during 1970-1977, 1978-1984, and 1985-1993 to mothers who ate more than 116 meals of GLSCF before pregnancy were, on average, 164 g lighter, 46 g heavier, and 134 g heavier, respectively, than children of mothers who ate no GLSCF before pregnancy (P trend=0.05). GLSCF-consuming mothers had higher serum PCB and DDE concentrations, but only increased DDE was associated with lower birth weight. The data suggest that fetal DDE exposure (as indicated by maternal serum DDE concentration) may decrease birth weight and that decreased birth weight effects associated with GLSCF consumption have decreased over time. PMID- 15533332 TI - Longitudinal study of babies born to mothers enrolled in a preconception prospective pregnancy study: study design and methodology, New York State Angler cohort study. AB - Persistent environmental chemicals such as organochlorine pesticides and polychlorinated biphenyls (PCBs) have been associated with alterations in fetal development and child health including subtle differences in developmental status. Previous prospective studies have ascertained prenatal or postnatal exposures but none have been designed to assess exposures at critical windows including preconception. To address this gap, we followed infants born to mothers recruited prior to conception in the New York State Prospective Pregnancy Study to assess feasibility issues including acceptability of a relatively invasive study protocol during the child's first 2 years of life. Longitudinal measurements on health, development, and growth were obtained from 53 live-born infants; 49 families consented to standardized in-home neurodevelopmental and psychosocial evaluations at 12 and 24 months of age. Nineteen participating parents consented to the collection of blood from infants for lead thyroid and PCB levels. Despite the intensive data collection protocol over 2 years coupled with the mothers having completed an intensive prospective pregnancy protocol, we found parents readily open to continued participation in a longitudinal study involving their children. Suggestions for conducting in-home assessments include use of a consistent contact nurse, comprehensive parent-friendly developmental assessment tools with some interim assessment by parent report, and periodic team visits. PMID- 15533333 TI - Predictors of organochlorines in New York City pregnant women, 1998-2001. AB - Organochlorine compounds (OCs) have been found widely in human tissues. However, levels have been rapidly declining since their virtual ban in the 1970s. We measured 1,1'-dichloro-2,2'-bis(4-chlorophenyl)ethylene (DDE), polychlorinated biphenyls (PCBs), and trans-nonachlor (TN) in 194 pregnant women in New York City and examined demographic and dietary predictors of their levels in serum. Serum OC levels were low (median microg/L: 0.64 DDE, 0.79 PCB); TN was largely below the level of detection (74%). In multivariate models, levels of OCs increased with age; DDE was higher in women not born in the US or Puerto Rico; PCB were higher in women who bought fresh fish and lower in those with higher body mass indices. PMID- 15533334 TI - Profiles of ortho-polychlorinated biphenyl congeners, dichlorodiphenyldichloroethylene, hexachlorobenzene, and Mirex among male Lake Ontario sportfish consumers: the New York State Angler cohort study. AB - Great Lakes sportfish consumption is a recognized human exposure source to environmental organochlorine compounds. Using data collected as part of the New York State Angler Cohort Study, 203 males were considered with regard to history of Lake Ontario sportfish consumption and sera levels of 57 ortho-substituted polychlorinated biphenyl (PCB) congeners, dichlorodiphenyldichloroethylene, hexachlorobenzene, and Mirex. Consumption of six species of highly contaminated Lake Ontario sportfish during the years 1980 to 1990 was considered. Exposure was weighted by average annual frequency of consumption, whole-fish PCB estimates, and meal size to determine a continuous index of exposure. This index was dichotomized to compare "consumers" to "nonconsumers" in several procedures. Recovery and lipid-adjusted serum values for PCB congener IUPAC Nos. 138+163 (rsp=0.3), 183 (rsp=0.3), 187 (rsp=0.4), and 188 (rsp=0.3) and Mirex (rsp=0.4) were significantly correlated with the index of fish consumption (P<0.001). A logistic regression model, using forward stepwise selection procedures (alpha in/out=0.002/0.004), was employed to account for high intercorrelations among organochlorine predictor variables. Mirex was the sole statistically significant predictor variable of "consumer/nonconsumer." A one-unit increase in log Mirex was associated with a 92.0% increase in the odds of having reported sportfish consumption (OR=1.92, 95%CI=1.43, 2.58). Age, body mass index, cigarette smoking, water/wildfowl consumption, and year of sampling were considered covariates. The results of this study support the premise that long-term dietary consumption of Lake Ontario sportfish, even at comparatively low levels, contributes significantly to the body burden of organochlorine compounds in western New York anglers and sportsmen. PMID- 15533335 TI - Blood mercury level and blood pressure among US women: results from the National Health and Nutrition Examination Survey 1999-2000. AB - Exposure to mercury has been linked to elevations in blood pressure (BP), though few data are available. We examined the cross-sectional relationship between blood mercury concentration and BP in a representative US sample of 1240 women, aged 16-49 years, from the National Health and Nutrition Examination Survey 1999 2000. We found no association overall between mercury and BP in multivariate models. We stratified our data by dietary fish intake (presumably reflecting the consumption of long-chain n-3 fatty acids that may reduce BP) resulting in 759 fish consumers and 481 non-fish consumers. We found that for each 1.3 microg/L (interquartile distance) increase in mercury, systolic BP significantly increased by 1.83 mm Hg (95% CI: 0.36, 3.30) among non-fish consumers. A similar pattern was seen for diastolic BP, although it was non-significant. While an adverse effect of mercury exposure at background levels on BP was not present overall, an adverse association was present among non-fish-consuming young and middle-aged women. PMID- 15533336 TI - Application of hair-mercury analysis to determine the impact of a seafood advisory. AB - Following an official recommendation in the Faroe Islands that women should abstain from eating mercury-contaminated pilot whale meat, a survey was carried out to obtain information on dietary habits and hair samples for mercury analysis. A letter was sent to all 1180 women aged 26-30 years who resided within the Faroes, and the women were contacted again 1 year later. A total of 415 women responded to the first letter; the second letter resulted in 145 repeat hair samples and 125 new responses. Questionnaire results showed that Faroese women, on average, consumed whale meat for dinner only once every second month, but the frequency and meal size depended on the availability of whale in the community. The geometric mean hair-mercury concentration at the first survey was higher in districts with available whale than in those without (3.03 vs. 1.88 microg/g; P=0.001). The mercury concentration also depended on the frequency of whale meat dinners and on the consumption of dried whale meat. The 36 women who did not eat whale meat at all had a geometric mean hair-mercury concentration of 1.28 microg/g. At the time of the second survey, the geometric mean had decreased to 1.77 microg/g (P<0.001), although whale was now available in all districts. In comparison with previously published data on hair-mercury concentrations in pregnant Faroese women, these results document substantially lower exposures as well as a further decrease temporally associated with the issue of a stricter dietary advisory. PMID- 15533337 TI - Hair mercury (signature of fish consumption) and cardiovascular risk in Munduruku and Kayabi Indians of Amazonia. AB - Fish is an important natural resource in the diet of inhabitants of the Amazon rain forest and a marker of its consumption (hair Hg) was used to compare selected cardiovascular risk parameters between tribes of Eastern Amazonia. Three Munduruku (Terra Preta, Kaburua, Cururu) villages and one Kayabi village at the banks of head rivers (Tapajos, Tropas, Kabitutu, Cururu, Curuzinho, Teles Pires) of the Tapajos Basin were studied in relation to fish Hg concentrations, mercury in hair (fish consumption) and erythrocytes, body mass index (height/weight, kg/cm2), and blood pressure. The mean fish Hg concentrations were higher in predatory (578.6 ng/g) than in nonpredatory species (52.8 ng/g). Overall only 26% of fish Hg concentrations were above 500 ng/g, and only 11% were above 1000 ng/g. There was no systematic trend in fish Hg concentrations from rivers with a history of gold-mining activities. The biomarker of fish consumption (hair Hg) was significantly associated with erythrocyte-Hg (r=0.5181; P=0.0001) and was significantly higher in Kayabi (12.7 microg/g) than in the Munduruku (3.4 microg/g). Biomarker-assessed fish consumption rate was higher in the Kayabi (110 g/day) than in the Munduruku villages (30 g/day). Although no significant differences in body mass index (BMI) were observed between tribes, there was a trend of lower increase in blood pressure with age among the higher fish consumers (Kayabi). Summary clinical evaluation did not detect neurologic complaints compatible with Hg intoxication (paraparesis, numbness, tremor, balancing failure), but endemic tropical diseases such as clinical history of malaria showed a high prevalence (55.4%). Fish is an abundant natural resource, important in the Indian diet, that has been historically consumed without perceived problems and can easily be traced through hair Hg. The exposure to freshwater fish monomethyl mercury is less of an issue than endemic infectious diseases such as malaria and lack of basic medical services. PMID- 15533338 TI - Fish consumption, advisory awareness, and hair mercury levels among women of childbearing age. AB - The Wisconsin Division of Public Health and the State of Maine Bureau of Health collaborated on a 12 state mercury awareness project. The primary goals of this initiative were to evaluate mercury sport fishing advisory awareness among women of childbearing age and assess the methylmercury exposure among this subpopulation. The project, which was conducted between December 1998 and December 1999, involved a randomized telephone survey of 3015 women of childbearing age and hair mercury analyses for a subset of 414 women. While 92% of these women had consumed fish or shellfish during the past year, less than one third (29%) of them had eaten any sport-caught fish. More than two thirds of the women who consumed sportfish were not aware that their state had issued fish consumption guidelines to protect against methylmercury exposure. Hair mercury levels ranged from 0.005 to 4.62 ppm and were positively correlated with recent fish consumption rates (P<0.0001). Women who ate sport-caught fish did not have significantly higher hair mercury levels than others (mean 0.51 vs. 0.48 ppm). Among women who ate sportfish, advisory awareness had no effect on their mercury exposure. Demographic indicators associated with higher hair mercury levels included residence in northeastern USA, marital status of married, college education, annual household income greater than $75,000, and Asian race. PMID- 15533339 TI - Clinical application of sperm-oocyte interaction tests in in vitro fertilization- embryo transfer and intracytoplasmic sperm injection programs. AB - OBJECTIVE: To review the clinical value of sperm-oocyte interaction tests for the diagnosis and management of infertility by standard IVF or intracytoplasmic sperm injection (ICSI). DESIGN: Review of recent publications on relationships among sperm-oocyte interaction tests, sperm characteristics, and results of IVF and determination of frequency of defective sperm-oocyte interaction in infertile men. MAIN OUTCOME MEASURE(S): Fertilization rates with IVF, sperm characteristics, sperm-zona pellucida (ZP) binding, ZP-induced acrosome reaction (AR), and sperm-ZP penetration. RESULT(S): Sperm defects associated with low sperm-ZP binding or impaired ZP-induced AR and sperm-ZP penetration are the major causes of failure of fertilization when all or most oocytes from a couple do not fertilize in standard IVF. There is a high frequency of defective sperm-ZP interaction in men with oligozoospermia (<20 x 10(6)/mL) and severe teratozoospermia (strict normal sperm morphology < or =5%). Sperm morphology correlates with sperm-ZP binding, and sperm concentration correlates with ZP induced AR in infertile men with sperm concentrations >20 x 10(6)/mL. Defective ZP-induced AR may cause infertility in up to 25% men with idiopathic infertility. These patients require ICSI despite the normal standard semen analyses. CONCLUSION(S): Sperm-oocyte interaction tests are useful for diagnosis of subtle sperm defects that cause infertility in men without severe abnormalities of semen analysis. Pre-IVF diagnosis of these sperm defects will assist in the clinical assignment of patients to treatment with either standard IVF or ICSI. PMID- 15533340 TI - Histological dating of timed endometrial biopsy tissue is not related to fertility status. AB - OBJECTIVE: To assess the ability of histological dating to discriminate between women of fertile and infertile couples. The utility of histological dating of endometrium in the evaluation of infertile couples is uncertain. DESIGN: Prospective multicenter study, with subjects randomly assigned to biopsy timing. Criterion standard for infertility was 12 months of unprotected, regular intercourse without conception and for fertility at least one live birth within 2 years. SETTING: University-based infertility practices. PATIENT(S): Volunteer subjects (847) recruited at 12 clinical sites participating in the National Institutes of Health-funded Reproductive Medicine Network. Inclusion criteria included ages 20-39 years, regular menstrual cycles, and no hormonal treatment or contraceptive use for 1 month before the study. Fertile controls were excluded if they had a history of infertility, recurrent pregnancy loss, or recent breastfeeding. INTERVENTION(S): Subjects underwent daily urinary LH testing. After detection of the LH surge, subjects were randomized to biopsy in the mid (days 21-22) or the late (days 26-27) luteal phase. Pathologists at each site estimated the cycle day based on standard criteria. For the primary analysis, an out-of-phase biopsy was defined as a greater than 2-day delay in the histological maturation of the endometrium. MAIN OUTCOME MEASURE(S): The proportion of out-of phase biopsies in fertile and infertile women was compared using logistic regression models with age at randomization as a covariate. Comparisons were also made between fertile vs. infertile at the midluteal or late luteal phase time points. RESULT(S): Biopsies were evaluated (301 mid and 318 late; N = 619). Out of-phase biopsy results poorly discriminated between women from fertile and infertile couples in either the midluteal (fertile: 49.4%, infertile: 43.2%) or late luteal phase (fertile: 35.3%, infertile 23.0%). Results did not substantially differ using alternative definitions of "out-of-phase" or standardized cycle day. CONCLUSION(S): Histological dating of the endometrium does not discriminate between women of fertile and infertile couples and should not be used in the routine evaluation of infertility. PMID- 15533341 TI - Absence of secretory endometrium after false-positive home urine luteinizing hormone testing. AB - OBJECTIVE: To examine the proportion of cases with proliferative endometrium on biopsies performed after positive home urine LH testing. DESIGN: Multicenter clinical trial of the usefulness of endometrial biopsy in the evaluation of infertility, with women from fertile and infertile couples randomly assigned to midluteal vs. late luteal phase endometrial sampling. SETTING: Twelve clinical sites of the National Institutes of Health/National Institute of Child Health and Human Development-sponsored Reproductive Medicine Network. PATIENT(S): All women in the study had regular menstrual cycles. Fertile volunteers who had delivered a live born infant within the past 2 years without medical intervention were recruited through advertisements at participating sites. Infertile women with regular cycles were recruited from the clinical practices of the sites' physicians. INTERVENTION(S): Interview, informed consent, subject-interpreted home urine LH testing, and endometrial biopsy in either the midluteal or late luteal phase. MAIN OUTCOME MEASURE(S): Proportion of cases with proliferative endometrium on biopsy. RESULT(S): In both fertile and infertile women, more than 7% of endometrial biopsies performed 7-13 days after a positive home urine LH test revealed proliferative endometrium. CONCLUSION(S): Patient interpretation of home urine LH test kits not uncommonly results in false-positive tests. Women planning menstrual cycle testing or procedures related to ovulation may benefit from additional confirmatory testing. PMID- 15533342 TI - Interobserver and intraobserver variability in the histological dating of the endometrium in fertile and infertile women. AB - OBJECTIVE: To assess effects of biopsy timing and fertility status on inter- and intraobserver variability in dating of the endometrium. DESIGN: Endometrial biopsy slides randomly selected from a multicenter study testing the utility of biopsy in the diagnosis of infertility were distributed to three gynecologic pathologists, who estimated cycle day using standard criteria. Readers were blinded to the purpose of the study, patient age, fertility status, or timing of biopsy relative to LH surge or next menses. SETTING: Multicenter academic research programs in reproductive medicine. PATIENT(S): Eighty-two women with proven fertility, 83 infertile patients. INTERVENTION(S): Endometrial biopsy during midluteal (days 21-22) or late (days 26-27) luteal phase. MAIN OUTCOME MEASURE(S): Intraclass correlation coefficient (ICC), kappa. RESULT(S): Overall agreement was excellent (ICC 0.88); addition of readings by local pathologists decreased ICC only slightly. In subgroup analyses, ICCs were lowest for infertile women during the midluteal phase (0.65 vs. 0.71 for fertile women in the midluteal phase, and 0.88-0.90 for both groups in the late luteal phase). Intraobserver reliability was excellent (0.9-0.99). Agreement for diagnoses of "out-of-phase" was only moderate, with kappa values between 0.4 and 0.6. CONCLUSION(S): Observer variability in dating the endometrium was greatest in infertile women during the window of implantation. PMID- 15533343 TI - Looking at the endometrial biopsy with evidence-based medicine. AB - The results of the study of endometrial biopsy for the diagnosis of infertility, which appear in this issue, are valid. Likelihood ratios and the receiver operating characteristic curves indicate that an out-of-phase test result is worse than useless; it is misleading. PMID- 15533344 TI - Grading a developmental continuum--elegy on the rise and fall of the endometrial biopsy. AB - Due to the original study design the frequency distribution of the slide categories sent to each of the three expert pathologists was essentially bimodal. The intraclass correlation coefficient for the 1-14 continuous categories reveals excellent agreement (0.82), whereas the kappa statistic for the dichotomized outcomes (>2 day lag) is poor (0.54). The discrepancy between the intraclass correlation coefficient and kappa values seems to be due to the bimodal distribution of the slide ratings. Some impartial mechanism was needed to make an unbiased selection of the slides used for endometrial dating. Imprecision in endometrial dating is due primarily to the limitations of the instrument and not the observer. Maturation stage of the endometrium is not critical for predicting a successful pregnancy. PMID- 15533345 TI - Endometrial biopsy: a test whose time has come. AB - The endometrial biopsy is still a precise screening tool for the evaluation of the infertile couple. PMID- 15533346 TI - Endometrial biopsy: a test whose time has come and gone. AB - To document ovulation in an infertility investigation, a timed endometrial biopsy is painful, expensive, invasive, requires precise cycle timing for proper interpretation, and cannot independently distinguish between fertile and infertile women. Urinary LH kit testing has a significant false-positive rate (approximately 7.6%). A midluteal serum progesterone is a much simpler method that compares favorably with either endometrial biopsy or LH kit testing. PMID- 15533347 TI - Endometrial biopsy should be abandoned as a routine component of the infertility evaluation. AB - Endometrial biopsies are of little value in the evaluation of infertile couples. Their usefulness in the evaluation of couples with repeated early pregnancy loss remains to be demonstrated. PMID- 15533351 TI - Add-back therapy in the treatment of endometriosis-associated pain. AB - OBJECTIVE: To determine the efficacy of GnRH analogue plus add-back therapy compared with GnRH analogue alone and estroprogestin in patients with relapse of endometriosis-associated pain. DESIGN: Randomized, controlled study. SETTING: University hospital. PATIENT(S): One hundred thirty-three women with relapse of endometriosis-related pain after previous endometriosis surgery. INTERVENTION(S): Forty-six women were treated with GnRH analogue plus add-back therapy, 44 women were given GnRH analogue alone, and 43 women received estroprogestin, for 12 months. MAIN OUTCOME MEASURE(S): Pain evaluation by a visual analogue scale, quality of life in treated patients according to the SF-36 questionnaire, and occurrence of adverse effects, including bone mass density loss, at pretreatment, after 6 months of treatment, at the end of treatment (12 months), and 6 months after discontinuation of treatment. RESULT(S): Patients treated either with GnRH analogue alone or GnRH analogue plus add-back therapy showed a higher reduction of pelvic pain, dysmenorrhea, and dyspareunia than patients treated with oral contraceptive, whereas patients treated with add-back therapy showed a better quality of life, as assessed with the SF-36 questionnaire, and adverse effects rate than the other two groups. CONCLUSION(S): Add-back therapy allows the treatment of women with relapse of endometriosis-associated pain for a longer period, with reduced bone mineral density loss, good control of pain symptoms, and better patient quality of life compared with GnRH analogue alone or oral contraceptive. PMID- 15533352 TI - Cyclooxygenase-2 expression in deep endometriosis and matched eutopic endometrium. AB - OBJECTIVE: To identify any relationship between cyclooxygenase-2 expression and the intensity of severe, endometriosis-related dysmenorrhea. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Patients with deep endometriosis. INTERVENTION(S): During surgery, paired samples of tissue representing deep endometriosis and eutopic endometrium were obtained from 46 patients. Control endometrial tissue samples were obtained from 34 fertile women who underwent laparoscopic tubal ligation or reversal of tubal sterilization. Pain assessment for dysmenorrhea was done with a 10-point linear analogue scale. MAIN OUTCOME MEASURE(S): The percentage of surface immunostained for Cox-2 was determined by an immunohistochemical technique. Relationships between pain score for dysmenorrhea and Cox-2 expression were analyzed. RESULT(S): Cox-2 expression was significantly higher in eutopic endometrial stromal cells from patients with deep endometriosis than in stroma from controls during the early, mid, and late secretory phases. In endometriosis patients, Cox-2 expression in eutopic endometrial stromal cells was significantly higher in women with more severe dysmenorrhea (pain score > or =7 vs. <7) during early and mid secretory phases. CONCLUSION(S): Elevated Cox-2 expression in stromal cells in eutopic endometrium from patients with deep endometriosis may play a role in severe, endometriosis related dysmenorrhea. PMID- 15533353 TI - High frequency of (TG)mTn variant tracts in the cystic fibrosis transmembrane conductance regulator gene in men with high semen viscosity. AB - OBJECTIVE: To evaluate a possible correlation between abnormal semen consistency and cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations and variant tracts. DESIGN: Study of CFTR mutations and variant tracts in men with high semen viscosity as compared with normospermic men. SETTING: University-based centers for andrology, clinical biochemistry, and cystic fibrosis. PATIENT(S): Forty-six male partners from infertile couples with sine causa high semen viscosity compared with 72 normospermic men. INTERVENTION(S): Semen sample collection. MAIN OUTCOME MEASURE(S): We obtained the (TG)mTn polymorphic tracts and a panel of 31 mutations of CFTR, semen viscosity, and semen variables. RESULT(S): The frequencies of the (TG)12 and T5 variant alleles were statistically significantly higher in men with high semen viscosity (17.4% and 7.6%, respectively) than in the normospermic control group (6.9% and 1.4%, respectively). The frequency of the genotypes carrying (TG)12 or T5 was statistically significantly higher in men with high semen viscosity (39.1%) than in the normospermic control group (16.7%). Four men with high semen viscosity showed the variant (TG)12T5 haplotype; one of these men presented variant tracts on both alleles. None of the normospermic controls showed a (TG)12T5 haplotype. CONCLUSION(S): Semen hyperviscosity could be considered a "minimal clinical expression" of cystic fibrosis; CFTR gene sequence variations may constitute the genetic basis for this disease. PMID- 15533354 TI - Serum antimullerian hormone/mullerian-inhibiting substance appears to be a more discriminatory marker of assisted reproductive technology outcome than follicle stimulating hormone, inhibin B, or estradiol. AB - OBJECTIVE: To test the hypothesis that the concentration of early follicular phase serum antimullerian hormone (AMH) or mullerian-inhibiting substance (MIS) is a useful marker of ovarian response and assisted reproductive technology (ART) outcome. DESIGN: Retrospective analysis of day 3 serum samples drawn before treatment. SETTING: Private ART program. PATIENT(S): One hundred nine consecutive serum samples from women younger than 42 years of age who were undergoing ovulation induction for IVF. INTERVENTION(S): Follicular aspiration for IVF after ovarian stimulation with FSH in a down-regulated cycle using GnRH-a treatment. MAIN OUTCOME MEASURE(S): Correlations between day 3 serum AMH/MIS, E2, FSH, inhibin B levels, and IVF outcome (i.e., number of retrieved mature oocytes, number and quality of embryos obtained, ongoing clinical pregnancy rates). Multivariate regression analysis on categorical data was performed to describe a predictive model of clinical pregnancy outcome. RESULT(S): Mean serum AMH/MIS value for clinical pregnancy (n = 38) was 2.4 ng/mL, in comparison to 1.1 ng/mL for those who did not become pregnant (n = 71). No differences were noted in mean values for day 3 FSH, inhibin B, or E2 between groups. Multivariate regression analysis demonstrated that day 3 serum AMH/MIS had the greatest independent contribution in predicting pregnancy outcomes. CONCLUSION(S): These data demonstrate a strong association between day 3 serum AMH/MIS level and IVF outcome in women younger than 42 years of age. Higher AMH/MIS concentrations are associated with a greater number of mature oocytes, a greater number of embryos, and ultimately a higher clinical pregnancy rate. Furthermore, AMH/MIS may offer greater prognostic value than other currently available serum markers of ART outcome. PMID- 15533355 TI - Number of good quality embryos on day 3 is predictive for both pregnancy and implantation rates in in vitro fertilization/intracytoplasmic sperm injection cycles. AB - OBJECTIVE: To evaluate the predictive value for implantation and pregnancy rates of the number of embryos that reach the eight-cell stage with less than 20% fragmentation (good quality embryos) on day 3. DESIGN: Prospective observational study. SETTING: Private IVF center. PATIENT(S): One hundred eighty-nine women undergoing IVF, or intracytoplasmic sperm injection with at least four zygotes, who have had three embryos transferred on day 3. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Pregnancy rate and implantation rate. RESULT(S): All patients (n = 189) had 3 embryos transferred. The patients were divided in five groups according to the number of good quality embryos on day 3 (0, 1, 2, 3, >3; respectively). For the five groups, the pregnancy rates were 2.9%, 33.3%, 45.5%, 39.4%, and 64.3%; the implantation rates were 2.0%, 17.0%, 20.5%, 19.3%, and 35.7%. No statistically significant differences in pregnancy rate or implantation rate were found for groups 1, 2, and 3 (when 1, 2, or 3 good quality embryos were present). The data were also analyzed after pooling these three groups. A statistically significant difference in pregnancy rate and implantation rate was found for group 0 (no good quality embryos), groups 1, 2, and 3 (1, 2, or 3 good quality embryos), and group >3 (>3 good quality embryos). CONCLUSION(S): The number of good quality embryos available on day 3 is a strong predictive value for both pregnancy rate and implantation rate. When good quality embryos are present on day 3, only two embryos should be transferred to minimize multiple pregnancies. PMID- 15533356 TI - Abnormal embryonic development diagnosed embryoscopically in early intrauterine deaths after in vitro fertilization: a preliminary report of 23 cases. AB - OBJECTIVE: To provide data about the phenotypic appearance of the embryo of early failed pregnancies after IVF. DESIGN: Clinical prospective descriptive study. SETTING: Tertiary care center. PATIENT(S): Twenty-three women who had conceived by IVF and had a missed abortion before 12 weeks of gestation. INTERVENTION(S): Embryoscopic examination of the embryo before curettage. Cytogenetic analysis of the chorionic villi by standard G-banding cytogenetic techniques or by comparative genomic hybridization in combination with flow cytometry analysis. MAIN OUTCOME MEASURE(S): Embryonic phenotype and karyotype were determined. RESULT(S): Twenty-one of 23 IVF embryos showed structural defects on embryoscopic examination. Seventeen of 23 specimens had a chromosomal abnormality. The majority were numerical aberrations such as monosomy X (2 cases). Trisomies for chromosomes 18 (one case), 16 (three cases), 15 (one case), 14 (two cases), 13 (one case), 12 (one case), 11 (one case), 10 (one case), 9 (one case), 8 (one case), and 3 (one case) were observed. A structural chromosome anomaly leading to a chromosomal trisomy was observed in one case. Aneuploidy explained the grossly abnormal embryonic development documented by embryoscopy in 15 of 21 cases. CONCLUSION(S): Aneuploidy is the major factor affecting normal embryonic development in missed abortions after IVF. Further investigation is needed to elucidate mechanisms that might prevent normal embryogenesis but evade detection by the cytogenetic techniques used in the present study. PMID- 15533357 TI - Long-term effects of tibolone on mammographic density. AB - OBJECTIVE: To investigate the long-term effect of tibolone on mammographic density. DESIGN: Open-label, nonrandomized study. SETTING: Academic research environment. PATIENT(S): Postmenopausal women. INTERVENTION(S): Tibolone was administered orally, mammograms were performed annually. MAIN OUTCOME MEASURE(S): Mammographic density according to the Wolfe classification, performed by two independent radiologists, both of whom were blinded to treatment group. RESULT(S): No statistically significant differences were found between the two groups in baseline demographic data. There were no statistically significant differences in mammographic density between the control and active groups at baseline or at 10 years. CONCLUSION(S): This pilot study shows that tibolone does not adversely alter the mammographic density of the breasts over 10 years of treatment. PMID- 15533358 TI - Heritability of menopausal age in mothers and daughters. AB - OBJECTIVE: To determine the heritability of age at natural menopause from mother daughter pairs. DESIGN: Two-generation families were selected to study heritability of menopausal age. SETTING: Subjects were drawn from a population based study. PATIENT(S): One hundred sixty-four mother-daughter pairs with a natural menopausal age. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The heritability of age at natural menopause estimated by a random-effects model. RESULT(S): A heritability of 44% (95% confidence interval, 36%, 50%) was estimated. CONCLUSION(S): This study confirms that heritable components largely determine the natural age at menopause. Reasons for the differences between heritability estimates based on sibling pairs and parent-child comparisons are discussed. PMID- 15533359 TI - Prevalence of polycystic ovaries and polycystic ovary syndrome in lesbian women compared with heterosexual women. AB - OBJECTIVE: To determine the prevalence of polycystic ovaries (PCO) and polycystic ovarian syndrome (PCOS) in lesbian women compared with heterosexual women undergoing fertility treatment. DESIGN: A prospective observational study. SETTING: The London Women's clinic and The Hallam Medical Center. Tertiary referral fertility setup. PATIENT(S): Six hundred eighteen women undergoing ovarian stimulation with or without IUI treatment between November 2001 and January 2003. Of these, 254 were self-identified as lesbians and 364 were heterosexual women. INTERVENTION(S): Baseline pelvic ultrasound examination and blood tests conducted to measure biochemical parameters such as FSH, LH, E(2), PRL, T, androstenedione (A), sex hormone-binding globulin (SHBG), and DHEAS were performed between day 2 and 3 of each woman's menstrual cycle. Tubal patency tests were performed by hysterosalpingography or laparoscopy. MAIN OUTCOME MEASURE(S): Biochemical parameters. RESULT(S): Eighty percent of lesbian women, compared with 32% of the heterosexual women, had PCO on pelvic ultrasound examination. Thirty-eight percent of lesbian women, compared with 14% of heterosexual women, had PCOS. There were no significant differences in the androgen concentrations between lesbian and heterosexual women with normal ovaries. However, lesbian women with PCO and PCOS had significantly higher androgen concentrations compared with heterosexual women with PCO and PCOS. Tubal disease was as common in lesbian women as in heterosexual women. CONCLUSION(S): There is a significantly higher prevalence of PCO and PCOS in lesbian compared with heterosexual women. Lesbian women with either PCO or PCOS had more pronounced hyperandrogenism than did heterosexual women with either PCO or PCOS. PMID- 15533360 TI - Comparison of follicular vascularization in normal versus polycystic ovaries during in vitro fertilization as measured using 3-dimensional power Doppler ultrasonography. AB - OBJECTIVE: The growth of the follicles induced by gonadotropins during IVF treatment is accompanied by physiologic angiogenesis, which is essential for the maturation of the oocytes. We describe the IVF-induced changes in the vascularization and compare normal with polycystic ovaries (PCOs). DESIGN: Prospective study. SETTING: Assisted reproductive unit at a university hospital. PATIENT(S): Sixty women who underwent IVF cycles. INTERVENTION(S): The ovarian vascularization was measured after pituitary down-regulation, FSH stimulation, and hCG-injection using three-dimensional power Doppler ultrasonography. MAIN OUTCOME MEASURE(S): The total ovarian vascularization was divided by the number of follicles. RESULT(S): After pituitary suppression, the ovarian vascularization/follicle was lower in polycystic ovary (PCO) patients. During ovarian stimulation, follicles in PCOs required a lesser amount of FSH to acquire the same level of vascularization than the follicles in normal ovaries. In addition, hCG induced an increase in the follicular vascularization in both normal and PCOs. The follicle count correlated with the total vascularized volume in the ovaries throughout the IVF cycle. CONCLUSION(S): Follicles in PCOs seem to be less vascularized than the follicles in normal ovaries after GnRH treatment but not after gonadotropin stimulation. It is possible that restricted blood supply to the follicles in PCO might be associated with the follicular arrest that is observed. We could confirm that follicles in PCO are more sensitive to gonadotropin stimulation than follicles in normal ovaries. PMID- 15533361 TI - Serum leptin levels correlate with obesity parameters but not with hyperinsulinism in women with polycystic ovary syndrome. AB - OBJECTIVE: To examine serum leptin concentrations in obese and lean patients with polycystic ovary syndrome (PCOS) to assess whether the changes in leptin levels are due to obesity or hormonal alterations. DESIGN: Controlled clinical study. SETTING: Academic research environment. PATIENT(S): Obese and lean women with PCOS. INTERVENTION(S): Blood samples were collected before and after food consumption. MAIN OUTCOME MEASURE(S): Serum leptin and insulin levels. RESULT(S): Serum leptin concentrations were significantly correlated with body mass index (r = 0.649) and also with HOMA (r = 0.535). However, after controlling for body mass index in a partial correlation analysis, no significant correlation was found between serum leptin levels and HOMA or hyperinsulinemia. While lean patients with PCOS had a significant correlation between leptin concentrations and obesity parameters, they did not show any significant correlation with insulin resistance parameters. CONCLUSION(S): Although leptin concentrations in women with PCOS correlate with insulin resistance/hyperinsulinemia, this is related only to obesity. PMID- 15533362 TI - Predictive value of the presence of an embryonic heartbeat for live birth: comparison of women with and without recurrent pregnancy loss. AB - OBJECTIVE: To determine the predictive value of an embryonic heart rate (EHR) for a live birth in women with and without a history of recurrent pregnancy loss (RPL). DESIGN: Prospective cohort study with concurrent controls. SETTING: A subspecialty clinic for couples with RPL. PATIENT(S): Three hundred pregnant women who previously had been diagnosed with RPL, followed prospectively compared with 300 age-, race-, and gestational age-matched pregnant control women. INTERVENTION(S): Transvaginal sonography between 6 to 8 weeks of gestation. MAIN OUTCOME MEASURE(S): The EHR was determined between 6 and 8 weeks of gestation by transvaginal sonography. Obstetrical history and current pregnancy outcome were evaluated. RESULT(S): Data were analyzed by using the two-tailed t test and Fisher's exact test. In women with RPL, an EHR predicted a successful live birth in 246 (82%) of 300, compared with 294 (98%) of 300 in control women. The mean (+/- SD) EHR from successful pregnancies in the control group (143.2 +/- 20.8 beats per minute) was significantly higher than the mean in women with a history of RPL (131.4 +/- 22.9 beats per minute). CONCLUSION(S): An EHR in women with RPL is associated with a live birth rate of 82% and is significantly lower than EHR in controls. Clinicians should use this information to counsel patients with RPL. PMID- 15533363 TI - Characterizing ectopic pregnancies that rupture despite treatment with methotrexate. AB - OBJECTIVE: To identify risk factors for tubal rupture among ectopic pregnancies treated with methotrexate (MTX). DESIGN: Retrospective case-control analysis. SETTING: An urban medical center. PATIENT(S): Eighty-one women diagnosed with an ectopic gestation treated with MTX: 19 patients experienced subsequent tubal rupture, and 62 patients experienced ectopic resolution. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Predictive variables including serial human chorionic gonadotropin (hCG) values. RESULT(S): The hCG incremental rate before as well as after MTX administration was positively associated with tubal rupture. HCG values prior to ectopic diagnosis that increased at least 66% over 48 hours and rising hCG values after treatment with methotrexate were independent predictors of tubal rupture. A disproportionate number (47%) of ectopic pregnancies that ruptured were located in the tubal isthmus. CONCLUSION(S): The hCG incremental rate both before and after MTX represents an independent risk factor for subsequent tubal rupture. Concentrations of hCG before ectopic diagnosis that increased at least 66% over 48 hours, or persistently rising hCG concentrations after treatment with MTX, may lower the threshold for surgical intervention. Implantation site may represent an unidentifiable risk factor for tubal rupture. PMID- 15533364 TI - Characterizing the endometrium in unexplained and tubal factor infertility: a multiparametric investigation. AB - OBJECTIVE: To characterize endometrial development in unexplained and tubal factor infertility. DESIGN: Prospective study of 20 women with unexplained infertility, 22 with tubal factor infertility, and 21 fertile controls in the midproliferative, periovulatory, and midluteal phases of the menstrual cycle. SETTING: Reproductive Medicine Department of St. Mary's Hospital, Manchester, United Kingdom. PATIENT(S): Women awaiting assisted conception. INVESTIGATION(S): Serum hormone assays, transvaginal ultrasound, Doppler, and midluteal endometrial biopsies. MAIN OUTCOME MEASURE(S): Serum levels of E2, P, and LH, endometrial ultrasound morphometry, uterine and subendometrial artery Doppler, and endometrial histology and biochemistry. RESULT(S): Women with unexplained infertility demonstrated significantly reduced uterine artery flow velocity in all phases, significantly elevated uterine and subendometrial artery impedance in the periovulatory and midluteal phases, and significantly reduced endometrial texture in the midproliferative phase. Women with tubal factor infertility demonstrated significantly reduced uterine artery flow velocity, without a concomitant increase in impedance, and significantly greater expression of endometrial glandular and luminal keratan sulphate. CONCLUSION(S): Unexplained infertility is associated with a profound impairment of endometrial perfusion that might be amenable to treatment by perfusion enhancers. Tubal factor infertility is associated with endometrial developmental defects that might be corrected by salpingectomy. Endometrial ultrasound and Doppler studies are likely to become a vital tool in the investigation of infertility. PMID- 15533365 TI - Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device. AB - OBJECTIVE: To test the feasibility of freezing intact human ovary using a passive cooling device. DESIGN: Prospective experimental study. SETTING: Academic research unit of the department of gynecology in a university hospital. PATIENT(S): Ovaries were obtained from three women who were between 29 and 36 years of age. INTERVENTION(S): Ovarian perfusion with cryoprotective solution and slow freezing with a cryofreezing container (5100 Cryo 1 degrees C Freezing Container; Nalgene, VWR, Belgium). Rapid thawing of ovaries by perfusion and bathing with decreased sucrose gradient. MAIN OUTCOME MEASURE(S): Viability of follicles, stromal cells, and vascular components was assessed with live-dead stains in freshly removed ovary, after cryoprotectant exposure before freezing, and after thawing. Histological morphology was assessed at these three different times. RESULT(S): The percentage of live follicles was 99.4% in fresh tissue, 98.1% after cryoprotectant exposure, and 75.1% after thawing. Viability assessment showed live stromal cells and small vessels after thawing. On histological evaluation, the morphology of follicles and cortical and medullar tissue was similar in all three groups. CONCLUSION(S): We described the cryopreservation, using an accessible protocol, of intact human ovary with its vascular pedicle and proved high survival rates of follicles, small vessels, and stromal cells and a normal histological structure in all the ovarian components after thawing using our protocol. PMID- 15533366 TI - Outpatient thermal balloon ablation of the endometrium. AB - OBJECTIVE: To determine the feasibility and potential efficacy of thermal balloon ablation of the endometrium in the outpatient setting without the need for general anesthesia or conscious sedation. DESIGN: Prospective observational study. SETTING: Outpatient hysteroscopy clinic in a university-affiliated teaching hospital. PATIENT(S): Fifty-three consecutively recruited women with menorrhagia that was unresponsive to medical treatment. INTERVENTION(S): Thermal balloon endometrial ablation using local anesthetic without conscious sedation. MAIN OUTCOME MEASURE(S): Procedure feasibility, change in menstrual symptoms, and patient satisfaction and quality of life (Menorrhagia Utility Scale and EuroQol) at 6-month follow-up. RESULT(S): Thermal balloon ablation was successfully completed in 50 (94%) of 53 women. The three failed procedures consisted of one case in which the woman could not tolerate the procedure because of severe discomfort, one case of equipment failure, and one case in which the balloon catheter could not be inserted into the uterine cavity. Completed outcome questionnaires were returned by 49 (98%) of 50 treated women. Improvement in menstrual loss was experienced by 39 (80%) of 49 women, and satisfaction with the outcome of treatment on menstrual symptoms was reported by 33 (67%) of 49 women. Significantly higher condition-specific quality-of-life scores were associated with treatment satisfaction. CONCLUSION(S): Thermal balloon ablation of the endometrium is feasible in the outpatient setting. Improvement in menstrual symptoms and satisfaction with the outcome of treatment appear to be comparable to published inpatient data. Further studies are required to determine the cost effectiveness of outpatient compared with inpatient thermal balloon therapy. PMID- 15533367 TI - Cannulation of a resistant internal os with the malleable outer sheath of a coaxial soft embryo transfer catheter does not affect in vitro fertilization embryo transfer outcome. AB - OBJECTIVE: To assess the impact of cannulation of a resistant cervical os with the outer malleable sheath of a double-lumen, soft ET catheter on IVF-ET outcomes. DESIGN: Retrospective cohort study. SETTING: University-based IVF center. PATIENT(S): One hundred forty-two patients undergoing 142 ETs. INTERVENTION(S): Trial ultrasound-guided ET at all transfers, leaving the malleable outer sheath in situ when the soft inner catheter could not negotiate the internal os. MAIN OUTCOME MEASURE(S): Implantation rate and clinical pregnancy rate. RESULT(S): In 102 ETs (71.8%), the soft inner sheath easily negotiated the internal os (group 1). Forty ETs (28.2%) required cannulation of resistant internal ora with the outer sheath of the trial catheter (group 2). Implantation rates (35% vs. 32% in groups 1 and 2, respectively) and clinical pregnancy rates (50% vs. 45%) were not significantly different between groups. Blood was present on the transfer catheter after ET more frequently in group 2 than in group 1 (55% vs. 15%); however, neither the implantation rate nor the clinical pregnancy rate were affected by the presence of blood. CONCLUSION(S): Cannulation of a resistant internal os by the malleable outer sheath and blood on the transfer catheter after ET do not have an adverse effect on implantation rate or clinical pregnancy rate. PMID- 15533368 TI - Transient abdominal ovariopexy for adhesion prevention in patients who underwent surgery for severe pelvic endometriosis. AB - OBJECTIVE: To assess adhesion reformation and subsequent fertility after a transient ovariopexy performed during severe pelvic endometriosis surgery. DESIGN: Retrospective study. SETTING: University hospital. PATIENT(S): Twenty young women who underwent severe pelvic endometriosis surgery. INTERVENTION(S): Unilateral or bilateral transient ovariopexy to the anterior abdominal wall was performed as the last step in the surgical procedure. Median duration of ovariopexy was 4 days. MAIN OUTCOME MEASURE(S): Adhesion reformation and subsequent fertility. RESULT(S): This well-tolerated procedure induced neither specific complication nor prolonged hospital stay. A second-look laparoscopy, performed in eight patients (40%), has shown a reduction of the occurrence, the extent, and the severity of ovarian adhesions. Two thirds of the suspended ovaries had no or smooth adhesions at second-look laparoscopy, even though all ovaries were initially adherent. Fifteen infertile women without male infertility factors tried actively to conceive after surgery. In this group of patients, four conceived spontaneously, and four conceived after IVF (total pregnancy rate = 53.3%). Seven patients delivered, and one pregnancy is ongoing. Median pregnancy delay was 11.5 months (range, 4-24 months). CONCLUSION(S): Transient ovariopexy appears to be a simple, safe, and effective technique in preventing postoperative adhesion reformation in severe pelvic endometriosis. PMID- 15533369 TI - Expectations for oocyte fertilization and embryo cleavage after whole sperm versus sperm head intracytoplasmic sperm injection. AB - OBJECTIVE: To compare oocyte fertilization and embryo development after intracytoplasmic sperm injection (ICSI) with a whole sperm vs. a sperm head. DESIGN: Retrospective study. SETTING: Hospital-based IVF practice. PATIENT(S): Fifty-three women undergoing 54 IVF-embryo transfer plus ICSI procedures between January 1999 and June 2002. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Oocyte fertilization, zygote cleavage, and embryo stage after 72 hours of culture. RESULT(S): A significantly higher fertilization rate was observed using whole sperm (72.2%) than when using sperm heads (56.4%). Zygote cleavage rates for whole sperm vs. sperm head ICSI were 96.4% and 92.7%, respectively. Embryo cell stage after 72 hours of culture for whole sperm vs. sperm head ICSI was 6.5 +/- 2.1 cells and 5.6 +/- 1.8 cells, respectively. Embryo grade at this same time point was not different (2.3 +/- 1.0 and 2.5 +/- 0.9, respectively). CONCLUSION(S): The ICSI using whole sperm produces superior fertilization rates compared to ICSI using sperm heads, but once oocytes were fertilized, zygote cleavage rates were not different between the two sperm sources. Oocytes injected with a whole sperm produced embryos of higher cell stage but equivalent quality compared to oocytes injected with sperm heads. Therefore, having only sperm heads for use in ICSI should not be a deterrent to using this procedure. PMID- 15533370 TI - High pregnancy rates can be achieved after freezing and thawing human blastocysts. AB - OBJECTIVE: To examine the results of a 3-year trial using blastocyst cryopreservation to limit multiple pregnancy and optimize overall pregnancy per cycle. DESIGN: Retrospective clinical evaluation of pregnancy rates after freezing and thawing human blastocysts. SETTING: Tertiary-care academic center. PATIENT(S): Seven hundred fifty-three different patients treated in 783 IVF cycles with blastocysts frozen from July 2000 to June 2003. INTERVENTION(S): Two thousand, two hundred fifty-nine blastocysts were frozen in cycles in which only blastocysts were cryopreserved (cycles with pronuclear stage oocytes or pre embryos also cryopreserved were excluded from the analysis). Of these, 628 (27.6%) were thawed in 218 cycles. MAIN OUTCOME MEASURE(S): Pregnancy rate per cycle with thaw. RESULT(S): Four hundred seventy-nine (76.3%) blastocysts survived thawing, and 440 (92.0%) were transferred after exhibiting evidence of survival (most commonly, blastocoele reexpansion). In cycles with a thaw, 211 (96.8%) of 218 underwent intrauterine transfer. An average of 2.09 blastocysts was transferred per replacement. One hundred twenty-five (59.2%) clinical pregnancies were established, which included 23 sets of twins and 5 triplet gestations. Two sets of monozygotic twins were identified after the replacement of a single thawed blastocyst (1.6%). The age of the patient at the time of cryopreservation (<37 years) was an important factor in the establishment of clinical and ongoing pregnancy. The mode of ovarian stimulation, replacement method, and whether blastocysts were frozen on day 5 or day 6 of development did not demonstrate clinical significance. CONCLUSION(S): Cryopreserved and thawed blastocysts demonstrated a similar potential for implantation when compared with fresh pre-embryos on day 3. On the basis of these results, the blastocyst stage of development appears to be optimal for clinical freeze-thaw trials. PMID- 15533371 TI - Appearance of uterine perforation by hysterosalpingography. AB - The appearance of a uterine perforation that occurred at the time of office hysteroscopy is shown via hysterosalpingogram and laparoscopy. PMID- 15533372 TI - An omphalopagus parasitic twin after intracytoplasmic sperm injection. AB - OBJECTIVE: To describe a first case of parasitic twin achieved after intracytoplasmic sperm injection (ICSI). DESIGN: Case report. SETTING: Perinatal center at a Japanese university hospital. PATIENT(S): A 30-year-old pregnant Japanese woman with a history of secondary infertility achieved after ICSI was diagnosed with an omphalopagus parasitic twin pregnancy by prenatal ultrasound at 28 weeks of gestation. INTERVENTION(S): A female infant was delivered by scheduled cesarean section at 30 weeks of gestation. The cesarean section was performed for dystocia. MAIN OUTCOME MEASURE(S): Brief review of parasitic twin and malformations in fetuses achieved after ICSI. RESULT(S): Resuscitation was not done because of the poor prognosis for both fetuses. Molecular analysis with informative genetic markers is consistent with monozygotic pregnancy. CONCLUSION(S): A careful ultrasound examination is indicated to detect additional anomalous findings in twin fetuses achieved after ICSI. PMID- 15533373 TI - Osseous metaplasia: case report and review. AB - OBJECTIVE: To discuss, through the experience of a case report and extensive literature review, the best practices for the diagnosis and treatment of osseous metaplasia, which is the cause of secondary infertility. DESIGN: Case report. SETTING: In vitro fertilization unit in Athens. PATIENT(S): A 40-year-old woman with a 10-year history of secondary infertility. INTERVENTION(S): Hysteroscopic diagnosis and removal of the bony fragment. MAIN OUTCOME MEASURE(S): Elimination of secondary infertility caused by osseous metaplasia. RESULT(S): After treatment, the woman underwent an IVF program and a healthy neonate was born with cesarean section. CONCLUSION(S): Hysteroscopy remains the best practice for the diagnosis and removal of endometrial ossifications, causing secondary infertility. PMID- 15533374 TI - Evaluation and treatment of familial globozoospermia in five brothers. AB - OBJECTIVE: To document the pathology of five siblings with complete globozoospermia and to report the effectiveness of repeated intracytoplasmic sperm injection (ICSI) treatment as therapy. DESIGN: Case report. SETTING: A tertiary center for assisted reproduction. PATIENTS(S): Five siblings with globozoospermia. INTERVENTION(S): Twenty cycles of repeated ICSI treatment. Detailed light and electron microscopy studies were performed on three of the globozoospermic brothers. MAIN OUTCOME MEASURE(S): Clinical pregnancy and live birth after ICSI treatment. RESULT(S): Light and electron microscopy showed that all spermatozoa had round heads and no acrosome. Of 129 metaphase-II oocytes injected, 49 fertilized normally, giving an overall fertilization rate of 38% (range, 0-100%). No pregnancies resulted from 13 cycles in brothers 1-3. Brothers 4 and 5 had three pregnancies after seven ICSI cycles, with one live birth and two first-trimester losses. CONCLUSION(S): Despite variable fertilization rates, pregnancies and a live birth can be achieved after repeated ICSI treatment cycles in globozoospermic siblings. No apparent pattern was manifest that reflected the true pathology or determined the outcome of ICSI treatment. PMID- 15533375 TI - Phosphatidylserine-dependent antiprothrombin antibodies are not useful markers for high-risk women with recurrent miscarriages. AB - To determine the possible association between antiprothrombin antibodies and conventional antiphospholipid antibody (aPL) 1 (1.0%), 2 (2.0%) and 17 (17.0%) were found to be positive for phosphatidylserine-dependent antiprothrombin antibody (aPS/PT) IgG, beta2glycoprotein I-dependent anticardiolipin antibody, and lupus anticoagulant by activated partial thromboplastin time (LA), respectively, in 100 recurrent aborters. Because patients with aPS/PT were included in the 17 with LA, we should not add aPS/PT measurement to routine testing in addition to conventional aPL for patients with recurrent miscarriage. PMID- 15533376 TI - Effects of short and long incubations on DNA fragmentation of testicular sperm. AB - DNA fragmentation in testicular sperm from men with obstructive azoospermia is increased by 4-hour and 24-hour incubations and after cryopreservation with the effect is intensified by post-thaw incubation. Testicular sperm for use in intracytoplasmic sperm injection (ICSI) should be injected without delay. PMID- 15533377 TI - A modified gonadotropin-releasing hormone (GnRH) antagonist protocol failed to increase clinical pregnancy rates in comparison with the long GnRH protocol. AB - The purpose of this prospective randomized study was to compare stimulation characteristics and IVF outcomes of the standard long GnRH agonist protocol for ovarian stimulation with a modified GnRH antagonist protocol. Starting GnRH antagonist in a flexible protocol according to the size of the leading follicle, with simultaneous augmentation of 75 IU recombinant FSH, failed to increase clinical pregnancy rates. PMID- 15533378 TI - Low-molecular-weight dextran infusion is more effective for the treatment of hemoconcentration due to severe ovarian hyperstimulation syndrome than human albumin infusion. AB - The most severe complication of ovarian hyperstimulation syndrome (OHSS) is thromboembolism, which is related to hemoconcentration. Dextran 40 infusion has greater effectiveness for the treatment of hemoconcentration due to OHSS than does human albumin infusion. PMID- 15533379 TI - Sperm cryopreservation and in vitro fertilization/intracytoplasmic sperm injection in men with congenital bilateral absence of the vas deferens: a success story. AB - In this study we evaluate the use of cryopreservation of sperm obtained at the time of surgical exploration in men with congenital bilateral absence of the vas deferens. We assess the impact of cryopreservation on pregnancy rates after IVF/intracytoplasmic sperm injection. Intraoperative cryopreservation of sperm at the time of microsurgical epididymal sperm aspiration in men with congenital bilateral absence of the vas deferens resulted in a 100% live delivery rate per couple, providing the highest pregnancy rates of any infertility treatment. PMID- 15533380 TI - Tyr204Phe and Val34Leu polymorphisms in two Brazilian ethnic groups and in patients with recurrent miscarriages. AB - This study investigated the prevalence of Tyr204Phe and Val34Leu polymorphisms in two Brazilian ethnic groups (171 Caucasians and 27 Blacks, and 117 men and 81 women) and in patients with recurrent miscarriages (RM) (86 women: 53 Caucasians and 33 Blacks). Study groups were matched to control groups by race and age. The prevalence of these polymorphisms did not differ between patients with RM and controls or between Caucasian and blacks, suggesting that these polymorphisms cannot be considered a risk factor for RM. PMID- 15533381 TI - Randomized, controlled trial of priming with 10,000 IU versus 20,000 IU of human chorionic gonadotropin in women with polycystic ovary syndrome who are undergoing in vitro maturation. AB - Preretrieval priming with 10,000 IU hCG can improve oocyte maturation rates in vitro for women undergoing in vivo maturation treatment, though the optimum dose is unknown. This prospective, randomized, controlled trial demonstrated no improvement in oocyte maturation rates with 20,000 IU of hCG compared with 10,000 IU of hCG and therefore no benefit of the higher dose. PMID- 15533382 TI - Mutational analysis of the melanocortin-4 receptor (MC4R) gene in children with premature pubarche and adolescent girls with hyperandrogenism. AB - To determine if variants in the melanocortin-4 receptor (MC4R) gene are associated with premature pubarche (PP) in children and hyperandrogenism (HA) in adolescent girls, we performed single-strand conformational polymorphism (SSCP) in 75 children (69 girls/six boys) with PP, 53 adolescent girls with HA, and 95 healthy adult control subjects. DNA sequence analysis of the conformers identified by SSCP revealed variants in six patients (two silent and one missense) and in none of the control subjects. PMID- 15533383 TI - Etiology of azoospermia in 100 consecutive nonvasectomized men. AB - History was taken systematically for 100 azoospermic, nonvasectomized men referred consecutively to a Danish fertility clinic. The men were examined by ultrasound, and their blood samples were analyzed for karyotype, Y microdeletions, and cystic fibrosis transmembrane conductance regulator gene mutations. In 29% of patients, the condition could be explained by genetic abnormalities; in 22%, by diseases or external influence; and in 27%, by former cryptorchidism. The azoospermic condition remained unexplained in only 22%. PMID- 15533384 TI - A polymorphism in the resistin gene promoter is associated with body mass index in women with polycystic ovary syndrome. AB - Resistin does not appear to be a major gene predisposing to polycystic ovary syndrome (PCOS). However, an association of a resistin variant with body mass index was found in women with PCOS, suggesting that resistin may be related to adiposity in PCOS. PMID- 15533385 TI - Low levels of pregnancy-associated plasma protein-A in asymptomatic women destined for miscarriage. AB - In an asymptomatic cohort, serum pregnancy-associated protein-A (PAPP-A) levels among women destined to miscarry were 14% of those seen with ongoing pregnancies. Levels were as low 3 weeks before diagnosis as on the day of diagnosis, suggesting that PAPP-A levels might predict future miscarriage. PMID- 15533387 TI - Nested polymerase chain reaction--not ready for gold standard status yet! PMID- 15533388 TI - Nested polymerase chain reaction--not ready for gold standard status yet! PMID- 15533389 TI - Implications of an irreversible procedure. PMID- 15533390 TI - Intravaginal medications. PMID- 15533392 TI - Intravaginal medications. PMID- 15533397 TI - hCG and time to intrauterine insemination? PMID- 15533394 TI - Limits of agreement? PMID- 15533398 TI - Adsorption of methylene blue on Cu(II)-exchanged montmorillonite. AB - Cu(2+)-exchanged montmorillonite (CEM) was prepared using the method of ion exchange reaction. The goal was to determine the adsorption of methylene blue (MB) onto montmorillonite and CEM. The results showed that treatment with Cu(2+) slightly reduced the adsorption of montmorillonite. Temperature, pH value, and ionic strength in the medium had greater or smaller effects on interaction between MB and the clay. The data obtained from MB adsorption onto the tested adsorbents followed the Langmuir and BET equations. Also, the adsorption processes were endothermic and spontaneous in nature. PMID- 15533399 TI - Roughness models for particle adhesion. AB - The effects of different surface roughness models on a previously developed van der Waals adhesion model were examined. The van der Waals adhesion model represented surface roughness with a distribution of hemispherical asperities. It was found that the constraints used to define the asperity distribution on the surface, which were determined from AFM scans, varied with scan size and thus were not constant for all surfaces examined. The greatest variation in these parameters occurred with materials that had large asperities or with materials where a large fraction of the surface was covered by asperities. These rough surfaces were modeled with fractals and also with a fast Fourier transform algorithm. When the model surfaces generated using the Fourier transforms are used in the adhesion model, the model accurately predicts the experimentally observed adhesion forces measured with the AFM. PMID- 15533400 TI - Characterization and determination of the thermodynamic and kinetic properties of p-CP adsorption onto organophilic bentonite from aqueous solution. AB - The characterization of tetraethylammonium bentonite and the adsorption of p chlorophenol (p-CP) onto organophilic bentonite (tetraethylammonium bentonite) was studied as a function of the solution concentration and temperature. The observed adsorption rates were found to fit first-order kinetics. The rate constants were calculated for temperatures ranging between 15.0 and 35.0 degrees C at constant concentration. The adsorption energy E and adsorption capacity q(m) for the phenolic compound adsorbing on organophilic bentonite were estimated using the Dubinin-Radushkevic equation. Thermodynamic parameters (Deltag(a), Deltah(a), Deltas(a)) were calculated by a new approximation from the isotherms of p-CP adsorption on organophilic bentonite. These isotherms were modeled according to Freundlich and Dubinin-Radushkevic adsorption isotherms. The amount of adsorption of p-chlorophenol on organophilic bentonite was found to be dependent on the relative energies of adsorbent-adsorbate, adsorbate-solvent, and adsorbate-adsorbate interactions. PMID- 15533401 TI - Adsorption and deformation phenomena at interaction of N2 and microporous carbon adsorbent. AB - Deformation of activated carbon adsorbent ACC (adsorbent obtained by removing Si from SiC) upon N(2) adsorption has been studied. To solve this problem, a dilatometer was used. Its main part was a line differential transformer, the core of which was connected to the adsorbent by means of a rod. Any changes in the adsorbent height caused a change in the core position in the transformer, which influenced the signal recorded from its secondary winding. These results were compared with the adsorption isotherms. High sensitivity of the dilatometric method has been shown. The dilatometer used allowed the measurement of absolute deformations in the range 1 x 10(-4) to 3 mm. PMID- 15533402 TI - The removal of heavy metal cations by natural zeolites. AB - In this study, the adsorption behavior of natural (clinoptilolite) zeolites with respect to Co(2+), Cu(2+), Zn(2+), and Mn(2+) has been studied in order to consider its application to purity metal finishing wastewaters. The batch method has been employed, using metal concentrations in solution ranging from 100 to 400 mg/l. The percentage adsorption and distribution coefficients (K(d)) were determined for the adsorption system as a function of sorbate concentration. In the ion exchange evaluation part of the study, it is determined that in every concentration range, adsorption ratios of clinoptilolite metal cations match to Langmuir, Freundlich, and Dubinin-Kaganer-Radushkevich (DKR) adsorption isotherm data, adding to that every cation exchange capacity metals has been calculated. It was found that the adsorption phenomena depend on charge density and hydrated ion diameter. According to the equilibrium studies, the selectivity sequence can be given as Co(2+) > Cu(2+) > Zn(2+) > Mn(2+). These results show that natural zeolites hold great potential to remove cationic heavy metal species from industrial wastewater. PMID- 15533403 TI - Adsorption of water vapor by poly(styrenesulfonic acid), sodium salt: isothermal and isobaric adsorption equilibria. AB - Air conditioning and dehumidifying systems based on sorption on solids are of great interest, especially in humid climates, because they allow reduction of thermal loads and use of chlorofluorocarbons. Previous studies have shown that hydrophilic polymers such as sulfonic polymers can have very high performance in water adsorption from air. The aim of this study was to characterize the water vapor adsorption properties of fully sulfonated and monosulfonated poly(styrenesulfonic acid), sodium salt, and to elucidate the mechanism of adsorption on these materials. Adsorption isotherms have been determined by TGA between 298 and 317 K for pressures ranging from 0.1 to 45 hPa. They have type II of the IUPAC classification and a small hysteresis loop between adsorption and desorption processes was observed only for the monosulfonated sample. Water content is up to 80% weight at 80% relative humidity. Adsorption isotherms have been well fitted with the FHH model. Adsorption-desorption isobars have been determined by TGA under 37 hPa in the temperature range 298-373 K. They show that these polymers can be completely regenerated by heating at 313 K under humidified air. No degradation of the adsorption properties has been observed after several regenerations. Adsorption enthalpies and entropies have been deduced from the Clapeyron equation and from DSC measurements. A good agreement was found. A mechanism of adsorption is proposed considering two kinds of adsorbate: bounded water in electrostatic interaction with functional groups and free water resulting from condensation. PMID- 15533404 TI - Adsorption isotherm models for basic dye adsorption by peat in single and binary component systems. AB - Colored effluents from textile industries are a problem in many rivers and waterways. Prediction of dye adsorption capacities is important in design considerations. The sorption of three basic dyes, namely Basic blue 3, Basic yellow 21, and Basic red 22, onto peat is reported. Equilibrium sorption isotherms have been measured for the three single-component systems. Equilibrium was achieved after 21 days. The experimental isotherm data were analyzed using Langmuir, Freundlich, Redlich-Peterson, Tempkin, and Toth isotherm equations. A detailed error analysis has been undertaken to investigate the effect of using different error criteria for the determination of the single-component isotherm parameters and hence obtain the best isotherm and isotherm parameters which describe the adsorption process. The linear transform model provided the highest R(2) regression coefficient with the Redlich-Peterson model. The Redlich-Peterson model also yielded the best fit to experimental data for all three dyes using the nonlinear error functions. An extended Langmuir model has been used to predict the isotherm data for the binary systems using the single component data. The correlation between theoretical and experimental data had only limited success due to competitive and interactive effects between the dyes and the dye-surface interactions. PMID- 15533405 TI - Simultaneous adsorption of copper ions and humic acid onto an activated carbon. AB - In this study, simultaneous adsorption of copper ions and humic acid (HA) from Aldrich onto an activated carbon is investigated. It is found that the HA adsorption in the absence of copper decreases as the pH is increased. It leads to a reduction of 34.7% in the specific surface area of carbon. There exists a critical concentration (CC) of HA for copper adsorption. At HA concentrations < CC, a decrease in copper adsorption is observed; however, the HA improves the adsorption at HA concentrations > CC. An increase in ionic strength can enhance the copper uptake; however, zinc and/or cobalt ions have an insignificant influence on copper adsorption. The adsorption is significantly increased by citric acid, whereas addition of EDTA slightly decreases the uptake. An intraparticle diffusion model is successfully used to describe the copper adsorption kinetics. PMID- 15533406 TI - Zn adsorption by different fractions of Galician soils. AB - To evaluate the contribution of organic matter, oxides, and clay fraction to Zn adsorption in six soils from Galicia (Spain), after soil characterization, adsorption isotherms were obtained by adding nine solutions containing between 20 and 500 mg L(-1) concentrations of Zn(NO(3))(2). Distribution coefficients were obtained from the data of adsorption isotherms. Zn adsorption isotherms corresponding to untreated soil and to the organic matter removed samples and organic matter and oxides removed samples were compared with curves pattern and adjusted to Langmuir and Freundlich empirical models. Untreated soils described L curves whereas when soils were deprived of any component, the curves described were S-type. Distribution coefficients allowed knowing the Zn adsorption capacity of the untreated soil, and of the organic matter, oxides, and clay fraction. Soil organic matter is the main component that affects Zn adsorption as long as soil pH is near neutrality. At acid pH, the oxides are the main component that affects Zn adsorption, although to a much smaller extent than the organic matter near neutral conditions. So soil pH is the main soil factor that determines Zn adsorption, before any other soil property. PMID- 15533407 TI - Adsorption of organic compounds onto polyelectrolyte immobilized-surfactant aggregates on cellulosic fibers. AB - The adsorption of anionic surfactants with different hydrophobic chain lengths onto cellulose fibers pretreated with a cationic polyelectrolyte has been investigated. Five steps are involved in the adsorption process, which was ascribed to the formation of monolayer and bilayer surfactant aggregates. Electrostatic interaction between the residual surface charges followed by hydrophobic interaction among the alkyl chains are considered the main factors in the adsorption process. The adsorption of the anionic surfactant was found to greatly enhance the retention of organic compounds onto the polyelectrolyte treated cellulose. The coadsorption phenomenon, which was dependent on the saturation level of the adsorbed surfactant, has been explained in terms of the accumulation of the organic solute on the hydrophobic core generated by the adsorbed layer. PMID- 15533408 TI - Equilibrium and kinetic studies of adsorption of phosphate onto ZnCl2 activated coir pith carbon. AB - Phosphate removal from aqueous solution was investigated using ZnCl(2)-activated carbon developed from coir pith, an agricultural solid waste. Studies were conducted to delineate the effect of contact time, adsorbent dose, phosphate concentration, pH, and temperature. The adsorption equilibrium data followed both Langmuir and Freundlich isotherms. Langmuir adsorption capacity was found to be 5.1 mg/g. Adsorption followed second-order kinetics. The removal was maximum in the pH range 3-10. pH effect and desorption studies showed that adsorption occurred by both ion exchange and chemisorption mechanisms. Adsorption was found to be spontaneous and endothermic. Effect of foreign ions on adsorption shows that perchlorate, sulfate, and selenite decreased the percent removal of phosphate. PMID- 15533409 TI - Rheology of asphalts modified with glycidylmethacrylate functionalized polymers. AB - Asphalt is known to be a colloidal suspension in which asphaltenes are covered by a stabilizing phase of polar resins and form complex micelles that are dispersed in the oily maltenic phase. In order to enhance its mechanical properties (e.g., in road paving), asphalts are often loaded with polymeric materials, thereby obtaining blends that can have different physical or chemical structures, depending on the composition of the added polymer. Asphalts modified by the addition of reactive ethylene terpolymers were prepared and their dielectric and rheological properties were measured both before and after a cure at high temperature. Even if it is not possible to determine the exact nature of the chemical interactions between asphalt and polymer, master curves obtained from dynamic data clearly show that during the cure the material tends to the behavior of a cross-linked network. PMID- 15533410 TI - Scaling properties of contact area of pressure-sensitive adhesives. AB - Two new devices were developed to provide accurate measurements of both the contact area and the tack strength of pressure-sensitive adhesives (PSAs). The first one is the "mechano-optical tack tester" (MOTT), which was designed to apply controlled contact pressure by means of a quartz prism probe, for determined contact times, onto the surfaces of PSA samples. The contact area is measured by the reflection of light at the quartz probe surface, which is in contact with the adhesive. The second device is an "acoustic contact tester" (ACT) that measures the tack strength and the contact area between a silicate glass and an adhesive by the reflection of an acoustic wave. Two ultrasonic sensors of different acoustic wavelengths have been built in order to study the scaling effects of the contact kinetics between an adhesive and the probe. MOTT and ACT experiments on a commercial PSA show that the contact area is the main parameter that governs the tack properties of the PSA. The experiments and the modeling point out that the contact area depends on the compression stress, the roughness, the thickness, and the Young's modulus of the PSA. However, comparison of contact kinetics from MOTT and ACT experiments show that the contact area is a subjective parameter that depends on the wavelength of the reflected beam used for its measurement. PMID- 15533412 TI - Aqueous latex/ceramic nanoparticle dispersions: colloidal stability and coating properties. AB - The effect of pH on the colloidal stability of aqueous dispersions containing antimony-doped tin oxide (ATO) or indium tin oxide (ITO) nanoparticles and poly(vinyl acetate-acrylic) copolymer (PVAc-co-acrylic) latex particles was investigated using experimental observations and Derjiaguin, Landau, Verwey and Overbeek (DLVO) theory. The microstructure, electrical properties and optical properties of composite coatings prepared from various dispersions were also studied. Zeta potential measurements revealed that the isoelectric point (IEP) of ATO nanoparticles was below pH 2.0, that of ITO nanoparticles was at pH approximately 6.0 and that of PVAc-co-acrylic latex was at pH approximately 2.0. ATO/PVAc-co-acrylic dispersions prepared at pH 3 were stable, but those prepared at pH 1.5 formed aggregates, which settled quickly with time. DLVO theory predictions are in accord with these results. Stable ITO/PVAc-co-acrylic dispersions are obtained at a pH of 3.0 and 11.0, but dispersions are not stable at a pH of 6.0, the IEP of ITO. At a pH of 3.0, DLVO results predict attraction between ITO particles and latex particles. Dispersion pH affected the microstructures and properties of ATO (or ITO)/PVAc-co-acrylic coatings. Suspensions that formed aggregates produced coatings with lower percolation thresholds and lower transparencies than those produced from stable suspensions. PMID- 15533411 TI - Adsorption of anionic dyes on chitosan beads. 1. The influence of the chemical structures of dyes and temperature on the adsorption kinetics. AB - In this work, chitosan beads were synthesized in acidic medium and cross-linked in 1% glutaraldehyde solution. The characterization of the materials using TG/DTG, XRD, and BET surface areas showed that the beads did not modify their characteristics after the cross-linking reaction. The cross-linked beads were utilized as adsorbents for the removal of the yellow-, blue-, and red-anionic reactive dyes from aqueous solutions at pH 2.0. Adsorption of the yellow-dye increased from 25 to 50 degrees C. However, adsorption of the blue-dye decreased from 25 to 50 degrees C. Interestingly, the adsorption of the red-dye decreased from 25 to 35 degrees C and increased from 45 to 50 degrees C. The kinetic data were evaluated using an Avrami kinetic model, where the parameter n was related to the determination of changes in the adsorption mechanisms. Adsorption data of the dyes in relation to the contact time, the chemical structures of the dyes, and temperature were presented and were discussed. PMID- 15533413 TI - Stability of aqueous TiO2 suspensions: influence of ethanol. AB - Dispersion of titania particles in water has been achieved thanks to the addition of two different compounds: (C(2)H(5))(4)NOH and Tiron. To compact particles using an electrophoretic deposition process, ethanol, up to 20 vol%, was added as cosolvent to prevent water electrolysis. Alcohol addition strongly affects zeta potential values of particles and increases suspension viscosity but without influence the surface charge. The more realistic interpretation is that integration of ethanol molecules into the water network moves the slipping plane away from the surface, reducing the repulsive potential between particles that leads to suspension destabilization. PMID- 15533414 TI - Force of a gas bubble on a foreign particle in front of a freezing interface. AB - We monitored the formation and development of a single gas bubble on the surface of a spherical particle of size 1.676 mm under unidirectional freezing and thawing (4.6-5.0 microm/s) and for the first time quantitatively estimated the force exerted on this particle by measuring the deformation of an attached elastic stick. The bubble would nucleate and grow on the particle surface closest to the ice front, while the force curve for a freezing-thawing cycle presented a hysteresis characteristic. This force was much greater than in the case without a bubble, and hence it dominated the engulfment process in the present freezing tests. The bubble force increased with increasing bubble size and was shown to be mainly attributable to the elastic force by the deformed bubble shape. Comments were made on the need to incorporate the role of bubbles in predicting the critical velocity to freeze a suspension with high dissolved gas content. PMID- 15533415 TI - Characterization of colloidal hematite particle shape and dispersion behavior. AB - An elliptically polarized light scattering (EPLS) technique for the size and shape characterization of ellipsoidal hematite particles is presented. The hematite particles are synthesized by aging aqueous FeCl(3) or Fe(NO(3))(3) solutions at 100 degrees C. Different reaction conditions create particles of aspect ratios between 1 (spherical) and 8 (elongated). Cross-sectional diameter and aspect ratio are observed as a function of reaction (aging) time. Growth of the elongated particles, as well as their fractal aggregation behavior, is characterized using EPLS and comparisons are made with photon correlation spectroscopy and TEM measurements. PMID- 15533416 TI - Surface-charging behavior of Zn-Cr layered double hydroxide. AB - A Zn-Cr layered double hydroxide (LDH) having the formula Zn(2)Cr(OH)(6)Cl(0.7)(CO(3))(0.15)2.1H(2)O was synthesized and characterized by powder X-ray diffraction, infrared spectroscopy, acid-base potentiometric titration, mass titration, electrophoretic mobility, and modeling of the electrical double layer. Adsorption of alizarin was also performed in order to show some particular features of the HDL. Net hydroxyl adsorption, which increases with increasing pH and decreasing supporting electrolyte concentration, takes place above pH 5. The electrophoretic mobility of the particles was always positive and it decreased when the pH was higher than 9. An isoelectric point of 12 could be estimated by extrapolating the data. The modified MUSIC model was used to estimate deprotonation constants of surface groups and different adsorption models were compared. Good fit of hydroxyl adsorption and electrophoresis could be achieved by considering both OH(-)/Cl(-) exchange at structural sites and proton desorption from surface hydroxyl groups. The modeling, in agreement with alizarin adsorption, indicates that most of the structural positive charge of the LDH is screened at the surface by exchanged anions and negatively charged surface groups. It also suggests that only structural charge sites initially neutralized by chloride ions are active for anion exchange. The remaining sites are blocked by carbonate and do not participate in the exchange. PMID- 15533417 TI - In vitro demineralization/remineralization cycles at human tooth enamel surfaces investigated by AFM and nanoindentation. AB - The early stages of enamel surface dissolution (erosion) are of high scientific relevance; however, little is known about if and to what extent these early stages, which result in the softening of the enamel surface, are reversible. The present study investigated the possible rehardening of surface softened enamel in two separate experiments in vitro, employing physiologically relevant demineralization and remineralization times and remineralizing solutions possessing chemical properties close to those of human saliva. Atomic force microscopy (AFM) based nanoindentation has been used to study the nanomechanical properties and the ultrastructure of polished enamel samples as affected by demineralization/remineralization cycles. In contradiction to previous studies employing microhardness techniques, no enamel rehardening has been observed after remineralization; however, the demineralization schemes used in the present study (short treatments with a citric acid buffer) are not comparable to the ones used in previous studies, making a comparison difficult. A mineral deposition after remineralization on softened enamel samples has been observed by AFM; however, a protective effect of this deposited layer could not be revealed. The exposure of softened enamel samples to either a remineralizing solution or a mineral water led to an improved acid resistance of these samples as shown in a demineralization/remineralization cycle. The present study has demonstrated that AFM based nanoindentation is a useful tool to investigate the demineralization and remineralization of surface softened enamel with high accuracy. PMID- 15533418 TI - Spreading and sorption of droplets on layered porous substrates. AB - In this paper spreading and sorption of a droplet on an anisotropic, layered porous substrate are investigated numerically. Flow in the saturated part of the porous material is governed by Darcy's law, assuming a sharp wetting front separating the saturated regions from the dry regions. Numerical results are presented for spreading and sorption of droplets in their dependence on the material and process parameters for axisymmetric configurations. Limiting cases of sorption into infinitely thick and very thin porous layers are considered. For an analytical sorption model for thin substrates fed by an infinite reservoir a correction term taking into account the flow resistance in the inlet region is derived and the consistence of the modified model with numerical and experimental results is shown. For two-layer substrates, numerical results on the influence of the layer permeabilities on the sorption kinetics are presented. PMID- 15533419 TI - Compositional changes in cement-stabilized waste during leach tests--comparison of SEM/EDX data with predictions from geochemical speciation modeling. AB - Cement-based stabilization/solidification (s/s) is a widely used treatment process for hazardous wastes containing toxic metals. The treated waste consists of a complex mixture of several solid phases produced by cement hydration reactions. Understanding and predicting the effects of leaching on these individual phases is essential for assessing the long-term immobilization of metal contaminants in s/s waste exposed to rain and groundwater. In this paper, particles of crushed Portland cement doped with copper, lead, and zinc nitrates were leached with nitric acid solutions maintained at constant pH in the range pH 4-7. Changes in solid composition at the microscopic scale were measured by scanning electron microscopy and energy dispersive X-ray spectrometry (SEM/EDX). The geochemical equilibrium model SOLTEQ-B, which accounts for the incongruent solubility behavior of calcium silicate hydrate, was used to simulate the compositional changes in the hydration gel for increasing extents of leaching. Measured concentrations of calcium, silicon, and sulfur were successfully predicted at all extents of leaching. Aluminum, lead, and zinc concentrations were also in good agreement with model predictions, except in the remineralization zones that form when metals solubilized in the outer regions of the s/s waste particles diffuse toward the interior and reprecipitate at higher pH. Copper was less accurately modeled at high extents of leaching. Accounting for the incorporation of contaminant metals into the cement hydration gel (in opposition to assuming the presence of individual metal hydroxides) is crucial for successfully predicting contaminant metal concentrations in the hydration gel at low and intermediate extents of leaching. PMID- 15533420 TI - Electrochemical properties of protoporphyrin IX zinc(II) films. AB - The electrochemical properties of protoporphyrin IX zinc(II) (ZnPP) films on indium-tin oxide (ITO) substrate have been studied for three types of films with different arrangements, which were an adsorbed film of ZnPP and LB films of ZnPP and its hybrid with hexadecyltrimethylammonium bromide. Cyclic voltammetry (CV) measurement showed that, as the adsorbed amount of ZnPP increases, an irreversible oxidation peak of ZnPP film is intensified. This reveals that electrochemical properties depend on the adsorbed amount rather than the orientation of porphyrin molecules. It was also supported from CV measurement and ultraviolet-visible absorption spectroscopy that porphyrins adsorbed on ITO substrate were desorbed after the single scan of potential. Additionally, photoresponse of these ZnPP films was investigated by photocurrent measurement. The photocurrent generation is due to carboxylic acid moieties but not ZnPP macrocycles. PMID- 15533421 TI - Determination of solid surface tension from particle-substrate pull-off forces measured with the atomic force microscope. AB - Atomic force microscopy (AFM) is capable of solid surface characterization at the microscopic and submicroscopic scales. It can also be used for the determination of surface tension of solids (gamma) from pull-off force (F) measurements, followed by analysis of the measured F values using contact mechanics theoretical models. Although a majority of the literature gamma results was obtained using either Johnson-Kendall-Roberts (JKR) or Derjaguin-Muller-Toporov (DMT) models, re analysis of the published experimental data presented in this paper indicates that these models are regularly misused. Additional complication in determination of gamma values using the AFM technique is that the measured pull-off forces have poor reproducibility. Reproducible and meaningful F values can be obtained with strict control over AFM experimental conditions during the pull-off force measurements (low humidity level, controlled and known loads) for high quality substrates and probes (surfaces should be free of heterogeneity, roughness, and contamination). Any probe or substrate imperfections complicate the interpretation of experimental results and often reduce the quality of the generated data. In this review, surface imperfection in terms of roughness and heterogeneity that influence the pull-off force are analyzed based upon the contact mechanics models. Simple correlations are proposed that could guide in selection and preparation of AFM probes and substrates for gamma determination and selection of loading conditions during the pull-off force measurements. Finally, the possibility of AFM measurements of solid surface tension using materials with rough surfaces is discussed. PMID- 15533422 TI - Rheological surface properties of C12DMPO solution as obtained from amplitude- and phase-frequency characteristics of an oscillating bubble system. AB - The experimental data on the surface rheological characteristics of dodecyl dimethyl phosphine oxide solutions obtained in a fully automatic oscillating bubble device under microgravity conditions in the frequency range 0.01-100 Hz are presented. The complex surface elasticity modulus is obtained form the amplitude- and phase-frequency characteristics of established pressure oscillations in a closed cell without calibration experiments by direct calculation of the necessary coefficients. The characteristics of the adsorption layers obtained from the elasticity modulus are in good agreement with adsorption isotherms and equations of state accounting for the intrinsic (2D) monolayer compressibility. PMID- 15533423 TI - Formation of gold nanoparticles in microreactor composed of helical peptide assembly in water. AB - A novel microreactor was prepared by self-assembly of an amphiphilic block copolymer composed of a hydrophobic helical peptide unit with a naphthyl group at the C terminal and a hydrophilic poly(ethylene glycol) unit. The copolymer formed a self-assembly in water, taking a vesicular structure. Noticeably, when the copolymer was dispersed in an Au(3+) aqueous solution, gold nanoparticles were formed without addition of any reducing reagent. The naphthyl groups, which are located at the inner surface of the vesicular assembly, promoted the reduction of Au(3+) ions with accompanying pH decrease. PMID- 15533424 TI - Effects of particle size on near-wall depletion in mono-dispersed colloidal suspensions. AB - In this work we investigate the change in particle concentration near a solid boundary for colloidal dispersions in pressure driven flow, commonly referred to as wall depletion. In particular we determine the effect of Peclet number on the strength and spatial extent of the depleted layer. The change in concentration near the solid boundary is measured with attenuated total reflection infrared (ATR-IR) spectroscopy described previously (P.J.A. Hartman Kok et al., J. Rheol. 46 (2002) 481). The method is capable of measuring the concentration of particles at distances ranging from 0.2 to 1.0 mum from the boundary. The suspensions investigated consisted of mono-dispersed polystyrene particles in water. Particles of four different sizes were used, with radius, a, of 30, 54, 105, and 197 nm. (The ratio H/a was in the range 2500-17,000 with H being the height of the flow cell.) This enabled us to measure the wall depletion effect over a wide range of Peclet numbers, ranging from 0.01 to 45. We found that wall depletion was not significant for Peclet numbers smaller than unity. Estimates of the wall slip layer thickness obtained from rheological experiments were consistent with the results obtained by ATR-IR spectroscopy. PMID- 15533425 TI - Electrophoresis of a concentrated dispersion of spherical particles covered by an ion-penetrable membrane layer. AB - The electrophoretic behavior of a concentrated dispersion of soft spherical particles is investigated theoretically, taking the effects of double-layer overlapping and double-layer polarization into account. Here, a particle comprises a rigid core and an ion-penetrable layer containing fixed charge, which mimics biocolloids and particles covered by artificial membrane layers. A cell model is adopted to simulate the system under consideration, and a pseudo spectral method based on Chebyshev polynomials is chosen for the resolution of the governing electrokinetic equations. The influence of the key parameters, including the thickness of the double layer, the concentration of particles, the surface potential of the rigid core of a particle, and the thickness, the amount of fixed charge, and the friction coefficient of the membrane layer of a particle on the electrophoretic behavior of the system under consideration is discussed. We show that while the result for the case of a dispersion containing rigid particles can be recovered as the limiting case of a dispersion containing soft particles, qualitative behaviors that are not present in the former are observed in the latter. PMID- 15533426 TI - Influence of the three-dimensional heterogeneous roughness on electrokinetic transport in microchannels. AB - Surface roughness has been considered as a passive means of enhancing species mixing in electroosmotic flow through microfluidic systems. It is highly desirable to understand the synergetic effect of three-dimensional (3D) roughness and surface heterogeneity on the electrokinetic flow through microchannels. In this study, we developed a three-dimensional finite-volume-based numerical model to simulate electroosmotic transport in a slit microchannel (formed between two parallel plates) with numerous heterogeneous prismatic roughness elements arranged symmetrically and asymmetrically on the microchannel walls. We consider that all 3D prismatic rough elements have the same surface charge or zeta potential, the substrate (the microchannel wall) surface has a different zeta potential. The results showed that the rough channel's geometry and the electroosmotic mobility ratio of the roughness elements' surface to that of the substrate, epsilon(mu), have a dramatic influence on the induced-pressure field, the electroosmotic flow patterns, and the electroosmotic flow rate in the heterogeneous rough microchannels. The associated sample-species transport presents a tidal-wave-like concentration field at the intersection between four neighboring rough elements under low epsilon(mu) values and has a concentration field similar to that of the smooth channels under high epsilon(mu) values. PMID- 15533427 TI - Comment on "Dynamic wetting by liquids of different viscosity," by T.D. Blake and Y.D. Shikhmurzaev. AB - We comment on a recent theory of dynamic wetting that is based directly upon a model for interface formation, introduced by Shikhmurzaev. We argue that the treatment of surface tension and its relaxation, inherent in the original model, is physically flawed. PMID- 15533429 TI - The interfacial torque on a partially submerged sphere. AB - For a heavy spherical particle floating at the surface of a liquid, we show that the resultant torque exerted on the particle about its center by the interfacial forces is always zero, regardless of the position and shape of the contact line, provided only that the interfacial tension and the contact angle are constant. Interfacial tension therefore cannot rotate a spherical particle about its center. PMID- 15533430 TI - Point of zero charge/isoelectric point of exotic oxides: Tl2O3. AB - The pristine point of zero charge of Tl(2)O(3) determined in the presence of NaNO(3) using the inert electrolyte titration method and confirmed by microelectrophoresis is at pH 7.9. PMID- 15533431 TI - Using xenon as a probe for dioxygen-binding sites in copper amine oxidases. AB - Potential dioxygen-binding sites in three Cu amine oxidases have been investigated by recording X-ray diffraction data at 1.7-2.2A resolution for crystals under a high pressure of xenon gas. Electron-density difference maps and crystallographic refinement provide unequivocal evidence for a number of Xe binding sites in each enzyme. Only one of these sites is present in all three Cu amine oxidases studied. Structural changes elsewhere in the protein molecules are insignificant. The results illustrate the use of xenon as a probe for cavities, in which a protein may accommodate a dioxygen molecule. The finding of a potential dioxygen-binding cavity close to the active site of Cu amine oxidases may be relevant to the function of the enzymes, since the formation of a transient protein-dioxygen complex is a likely step in the catalytic mechanism. No evidence was found for xenon binding in a region of the molecule that was previously identified in two other Cu amine oxidases as a potential transient dioxygen-binding site. PMID- 15533432 TI - GalR represses galP1 by inhibiting the rate-determining open complex formation through RNA polymerase contact: a GalR negative control mutant. AB - GalR represses the galP1 promoter by a DNA looping-independent mechanism. Equilibrium binding of GalR and RNA polymerase to DNA, and real-time kinetics of base-pair distortion (isomerization) showed that the equilibrium dissociation constant of RNA polymerase-P1 closed complexes is largely unaffected in the presence of saturating GalR, indicating that mutual antagonism (steric hindrance) of the regulator and the RNA polymerase does not occur at this promoter. In fluorescence kinetics with 2-AP labeled P1 DNA, GalR inhibited the slower of the two-step base-pair distortion process. We isolated a negative control GalR mutant, S29R, which while bound to the operator DNA was incapable of repression of P1. Based on these results and previous demonstration that repression requires the C-terminal domain of the alpha subunit (alpha-CTD) of RNA polymerase, we propose that GalR establishes contact with alpha-CTD at the last resolved isomerization intermediate, forming a kinetic trap. PMID- 15533433 TI - The structure of the cytoplasmic domain of EpsL, an inner membrane component of the type II secretion system of Vibrio cholerae: an unusual member of the actin like ATPase superfamily. AB - The type II secretion system (T2SS) is used by several Gram-negative bacteria for the secretion of hydrolytic enzymes and virulence factors across the outer membrane. In these secretion systems, a complex of 12-15 so-called "Gsp proteins" spans from a regulatory ATPase in the cytoplasm, via several signal or energy transducing proteins in the inner membrane and the pseudopilins in the periplasm, to the actual pore in the outer membrane. The human pathogen Vibrio cholerae employs such an assembly, called the Eps system, for the export of its major virulence factor, cholera toxin, from its periplasm into the lumen of the gastro intestinal tract of the host. Here, we report the atomic structure of the major cytoplasmic domain of the inner membrane-spanning EpsL protein from V. cholerae. EpsL is the binding partner of the regulatory ATPase EpsE as well as of EpsM and pseudopilins, and is therefore a critical link between the cytoplasmic and the periplasmic part of the Eps-system. The 2.7A resolution structure was determined by a combination of Se-Met multiple anomalous dispersion (MAD) and multiple isomorphous replacement with anomalous scattering (MIRAS) phasing methods. The 28kDa cytoplasmic domain of EpsL (cyto-EpsL) consists of three beta-sheet-rich domains. With domains I and III similar to the RNaseH-fold, cyto-EpsL unexpectedly shows structural homology with the superfamily of actin-like ATPases. cyto-EpsL, however, is an unusual member of this superfamily as it misses the canonical actin domains 1B and 2B, which are common yet variable in this superfamily. Moreover, cyto-EpsL has an additional domain II, which has the topology of an SHS2-fold module. Within the superfamily this fold module has been observed only for domain 1C of the cell division protein FtsA, in which it mediates protein-protein interactions. This domain II displays great flexibility and contributes to a pronounced negatively charged canyon on the surface of cyto EpsL. Functional data as well as structural homology and sequence conservation suggest that domain II interacts with EpsE, the major cytoplasmic binding partner of EpsL. PMID- 15533434 TI - Recognition of internal cleavage sites by retroviral RNases H. AB - The RNase H activity of reverse transcriptase is essential to complete retroviral replication. Many studies have characterized how reverse transcriptase associates with recessed and exposed DNA 3' ends or RNA 5' ends to position the RNase H domain for cleavage, but little is known about how a nick might affect RNase H cleavages, or how RNase H carries out internal cleavages, which do not require positioning by a nucleic acid end. We have addressed these issues using model hybrid substrates and the reverse transcriptases of Moloney murine leukemia virus (M-MuLV) and human immunodeficiency virus type 1 (HIV-1). Our results show that a nick separating an upstream RNA and a downstream RNA annealed to DNA is essentially ignored by RNase H, indicating that the RNA 5' end at a nick is not sufficient to position 5' end-directed cleavages. Cleavage sites that are located close to the 5' end of the downstream RNA are not recognized in the absence of the upstream RNA, and the 5' ends of the shorter upstream RNAs enhance cleavage at these sites. The recognition of an internal cleavage site depends on local sequence features found both upstream and downstream of the cleavage site, designated as the -1/+1 position. By analyzing the nucleotide frequencies in the sequence surrounding strong internal cleavage sites, preferred nucleotides have been identified in the flanking sequences spanning positions -14 to +1 for HIV-1 and -11 to +1 for M-MuLV. These data reveal that general degradation of the retroviral genome after minus-strand synthesis can occur through sequence specific cleavages. PMID- 15533435 TI - Analysis of the context dependent sequence requirements of active site residues in the metallo-beta-lactamase IMP-1. AB - The metallo-beta-lactamase IMP-1 catalyzes the hydrolysis of a broad range of beta-lactam antibiotics to provide bacterial resistance to these compounds. In this study, 29 amino acid residue positions in and near the active-site pocket of the IMP-1 enzyme were randomized individually by site-directed mutagenesis of the corresponding codons in the bla(IMP-1) gene. The 29 random libraries were used to identify positions that are critical for the catalytic and substrate-specific properties of the IMP-1 enzyme. Mutants from each of the random libraries were selected for the ability to confer to Escherichia coli resistance to ampicillin, cefotaxime, imipenem or cephaloridine. The DNA sequence of several functional mutants was determined for each of the substrates. Comparison of the sequences of mutants obtained from the different antibiotic selections indicates the sequence requirements for each position in the context of each substrate. The zinc chelating residues in the active site were found to be essential for hydrolysis of all antibiotics tested. Several positions, however, displayed context dependent sequence requirements, in that they were essential for one substrate(s) but not others. The most striking examples included Lys69, Asp84, Lys224, Pro225, Gly232, Asn233, Asp236 and Ser262. In addition, comparison of the results for all 29 positions indicates that hydrolysis of imipenem, cephaloridine and ampicillin has stringent sequence requirements, while the requirements for hydrolysis of cefotaxime are more relaxed. This suggests that more information is required to specify active-site pockets that carry out imipenem, cephaloridine or ampicillin hydrolysis than one that catalyzes cefotaxime hydrolysis. PMID- 15533436 TI - Translesion synthesis past 2'-deoxyxanthosine, a nitric oxide-derived DNA adduct, by mammalian DNA polymerases. AB - Cellular DNA is damaged by nitric oxide (NO), a multifunctional bioregulator and an environmental pollutant that has been implicated in diseases associated with cancer and chronic inflammation. 2'-Deoxyxanthosine (dX) is a major NO-derived DNA lesion. To explore the mutagenic potential of dX, a 38-mer oligodeoxynucleotide ((5')CATGCTGATGAATTCCTTCXCTTCTTTCCTCTCCCTTT) modified site specifically with dX at the X position was prepared post-synthetically and used as a DNA template in primer extension reactions catalyzed by calf thymus DNA polymerase (pol) alpha and human DNA pol beta, eta, and kappa. Primer extension reactions catalyzed by pol alpha or beta in the presence of four dNTPs were retarded at the dX lesion while pol eta and kappa readily bypassed the lesion. The fully extended products were analyzed to quantify the miscoding specificity and frequency of dX using two-phase polyacrylamide gel electrophoresis (PAGE). With pol alpha, eta and kappa, incorrect dTMP was preferentially incorporated opposite the lesion, along with lesser amounts of dCMP, the correct base. When pol beta was used, direct incorporation of correct dCMP was primarily observed, accompanied by small amounts of misincorporation of dTMP, dAMP and dGMP. Steady state kinetic analyses supported the results obtained from the two-phase PAGE assay. dX is a miscoding lesion capable of preferentially generating G-->A mutations. The miscoding frequency varied depending on DNA polymerase used. PMID- 15533437 TI - Alu element mutation spectra: molecular clocks and the effect of DNA methylation. AB - In primate genomes more than 40% of CpG islands are found within repetitive elements. With more than one million copies in the human genome, the Alu family of retrotransposons represents the most successful short interspersed element (SINE) in primates and CpG dinucleotides make up about 20% of Alu sequences. It is generally thought that CpG dinucleotides mutate approximately ten times faster than other dinucleotides due to cytosine methylation and the subsequent deamination and conversion of C-->T. However, the disparity of Alu subfamily age estimations based upon CpG or non-CpG substitution density indicates a more complex relationship between CpG and non-CpG substitutions within the Alu elements. Here we report an analysis of the mutation patterns for 5296 Alu elements comprising 20 subfamilies. Our results indicate a relatively constant CpG versus non-CpG substitution ratio of approximately 6 for the young (AluY) and intermediate (AluS) Alu subfamilies. However, a more complex non-linear relationship between CpG and non-CpG substitutions was observed when old (AluJ) subfamilies were included in the analysis. These patterns may be the result of the slowdown of the neutral mutation rate during primate evolution and/or an increase in the CpG mutation rate as the consequence of increased DNA methylation in response to a burst of retrotransposition activity approximately 35 million years ago. PMID- 15533438 TI - PMA induces the MUC5AC respiratory mucin in human bronchial epithelial cells, via PKC, EGF/TGF-alpha, Ras/Raf, MEK, ERK and Sp1-dependent mechanisms. AB - Asthma and chronic obstructive pulmonary disease are highly prevalent and economically important inflammatory airway diseases associated with mucus hypersecretion. Considerable additional morbidity and mortality are related to acute exacerbations, which are associated with further mucus hypersecretion. MUC5AC is a prominent airway mucin; however, the signalling pathways regulating MUC5AC hypersecretion are not fully characterised. We investigated the signalling pathway regulating phorbol 12-myristate 13-acetate (PMA)-induced MUC5AC gene and protein expression in human respiratory epithelial cells. Using NCI-H292 cells, we demonstrated that treatment with PMA increased production of total and MUC5AC specific mucin proteins. This increase was dependent on de novo MUC5AC gene transcription. We identified a short, proximal region of the MUC5AC promoter essential for this activity containing three specificity protein (Sp) 1 transcription factor-binding sites and a single CACCC site. By chemical inhibition, site-directed promoter mutagenesis and electrophoretic mobility-shift assay (EMSA), we demonstrated that PMA induced proteins binding to all three Sp1 sites and that they were all required for full induction of MUC5AC promoter activity. We then demonstrated a Ras-Raf-MEK/ERK signalling pathway was exclusively activated upstream of Sp1 activating the promoter and confirmed the requirement for matrix metalloproteinase activation leading to a ligand-dependent activation of the epidermal growth factor receptor. Finally, we demonstrated that activation of the novel protein kinase C isoforms delta and theta; was required upstream of the metalloproteinase activation. We have characterised a signalling pathway regulating PMA induction of MUC5AC. Studies such as this identify key signalling intermediates as targets for pharmacological intervention to treat mucus hypersecretion. PMID- 15533439 TI - Uch2/Uch37 is the major deubiquitinating enzyme associated with the 26S proteasome in fission yeast. AB - Conjugation of proteins to ubiquitin plays a central role for a number of cellular processes including endocytosis, DNA repair and degradation by the 26S proteasome. However, ubiquitination is reversible as a number of deubiquitinating enzymes mediate the disassembly of ubiquitin-protein conjugates. Some deubiquitinating enzymes are associated with the 26S proteasome contributing to and regulating the particle's activity. Here, we characterise fission yeast Uch2 and Ubp6, two proteasome associated deubiquitinating enzymes. The human orthologues of these enzymes are known as Uch37 and Usp14, respectively. We report that the subunit Uch2/Uch37 is the major deubiquitinating enzyme associated with the fission yeast 26S proteasome. In contrast, the activity of Ubp6 appears to play a more regulatory and/or structural role involving the proteasome subunits Mts1/Rpn9, Mts2/Rpt2 and Mts3/Rpn12, as Ubp6 becomes essential when activity of these subunits is compromised by conditional mutations. Finally, when the genes encoding Uch2/Uch37 and Ubp6 are disrupted, the cells are viable without showing obvious signs of impaired ubiquitin dependent proteolysis, indicating that other deubiquitinating enzymes may remedy for the redundancy of these enzymes. PMID- 15533440 TI - A measure for the angle between projections based on the extent of correlation between corresponding central sections. AB - A pre-condition for the ab initio assignment of Euler angles to a set of projections from an asymmetric object is that at least three of the available projections correspond to rotations about different axes. For symmetric objects this condition may be relaxed. There are some applications of single-particle electron microscopy, such as the reconstruction of filamentous macromolecular assemblies, where all available projections more-or-less correspond to rotations about a common rotation axis making it difficult to satisfy this condition. Here, a method has been developed to overcome this problem, based on the fact that the correlation between two central sections of the Fourier transform of a compact object will not be limited to an infinitesimal central line but will have a finite extent, which is related to the angle between the corresponding projections. Projections from model filaments, with different degrees of rotational symmetry about the long axis, have been used to test the methodology. The results show that angle determination is robust down to signal-to-noise ratios as low as 2 and that, in general, the error decreases as the degree of symmetry increases. The method has been used to assign angles to a set of negatively stained muscle thick filament projections to obtain an initial 3D reconstruction. The main features of the projections are seen to be faithfully reproduced in the reprojections from the reconstruction. A real-space adaptation of this method is also discussed. PMID- 15533441 TI - Simulation of the substrate cavity dynamics of quercetinase. AB - Molecular dynamics (MD) simulations have been performed on quercetin 2,3 dioxygenase (2,3QD) to study the mobility and flexibility of the substrate cavity. 2,3QD is the only firmly established Cu-containing dioxygenase known so far. It catalyses the breakage of the O-heterocycle of flavonols. The substrates occupy a shallow and overall hydrophobic cavity proximal to the metal centre of the homo-dimeric enzyme. The linker connecting the C-terminal and N-terminal domains in the monomer is partly disordered in the crystal structure and part of it forms a flexible lid at the entrance of the substrate cavity. This loop has been tentatively assigned a role in the enzyme mechanism: it helps lock the substrate into place. The dynamics of this loop has been investigated by MD simulation. The initial coordinates were taken from the crystal structure of 2,3QD in the presence of the substrate kaempferol (KMP). After equilibration and simulation over 7.2ns the substrate was removed and another equilibration and simulation of 7.2ns was performed. The results show that the structures of the free enzyme as well as of the enzyme-substrate complex are stable in MD simulation. The linker shows strongly enhanced mobility in the loop region that is close to the entrance to the substrate cavity (residues 154-169). Movement of the loop takes place on a timescale of 5-10ns. To confirm the conclusions about the loop dynamics drawn from the 7.2ns simulation, the simulation was extended with another 8ns. When substrate binds into the cavity the loop orders remarkably, although mobility is retained by residues 155-158. Some regions of the loop (residues 154-160 and 164-176) move over a considerable distance and approach the substrate closely, reinforcing the idea that they lock the substrate in the substrate cavity. The enthalpic component of the interaction of the loop with the protein and the KMP appears to favour the locking of the substrate. Two water molecules were found immobilised in the cavity, one of which exhibited rotation on the picosecond timescale. When the substrate is removed, the empty cavity fills up with water within 200ps. PMID- 15533442 TI - Acid-induced unfolding of the amino-terminal domains of the lethal and edema factors of anthrax toxin. AB - The two enzymatic components of anthrax toxin, lethal factor (LF) and edema factor (EF), are transported to the cytosol of mammalian cells by the third component, protective antigen (PA). A heptameric form of PA binds LF and/or EF and, under the acidic conditions encountered in endosomes, generates a membrane spanning pore that is thought to serve as a passageway for these enzymes to enter the cytosol. The pore contains a 14-stranded transmembrane beta-barrel that is too narrow to accommodate a fully folded protein, necessitating that LF and EF unfold, at least partly, in order to pass. Here, we describe the pH-dependence of the unfolding of LF(N) and EF(N), the 30kDa N-terminal PA-binding domains, and minimal translocatable units, of LF and EF. Equilibrium chemical denaturation studies using fluorescence and circular dichroism spectroscopy show that each protein unfolds via a four-state mechanism: N<-->I<-->J<-->U. The acid-induced N- >I transition occurs within the pH range of the endosome (pH 5-6). The I state predominates at lower pH values, and the J and U states are populated significantly only in the presence of denaturant. The I state is compact and has characteristics of a molten globule, as shown by its retention of significant secondary structure and its ability to bind an apolar fluorophore. The N-->I transition leads to an overall 60% increase in buried surface area exposure. The J state is expanded significantly and has diminished secondary structure content. We analyze the different protonation states of LF(N) and EF(N) in terms of a linked equilibrium proton binding model and discuss the implications of our findings for the mechanism of acidic pH-induced translocation of anthrax toxin. Finally, analysis of the structure of the transmembrane beta-barrel of PA shows that it can accommodate alpha-helix, and we suggest that the steric constraints and composition of the lumen may promote alpha-helix formation. PMID- 15533443 TI - Direct observation of Abeta amyloid fibril growth and inhibition. AB - Amyloid fibril formation is a phenomenon common to many proteins and peptides, including amyloid beta (Abeta) peptide associated with Alzheimer's disease. To clarify the mechanism of fibril formation and to create inhibitors, real-time monitoring of fibril growth is essential. Here, seed-dependent amyloid fibril growth of Abeta(1-40) was visualized in real-time at the single fibril level using total internal reflection fluorescence microscopy (TIRFM) combined with the binding of thioflavin T, an amyloid-specific fluorescence dye. The clear image and remarkable length of the fibrils enabled an exact analysis of the rate of growth of individual fibrils, indicating that the fibril growth was a highly cooperative process extending the fibril ends at a constant rate. It has been known that Abeta amyloid formation is a stereospecific reaction and the stability is affected by l/d-amino acid replacement. Focusing on these aspects, we designed several analogues of Abeta(25-35), a cytotoxic fragment of Abeta(1-40), consisting of l and d-amino acid residues, and examined their inhibitory effects by TIRFM. Some chimeric Abeta(25-35) peptides inhibited the fibril growth of Abeta(25-35) strongly, although they could not inhibit the growth of Abeta(1-40). The results suggest that a more rational design of stereospecific inhibitors, combined with real-time monitoring of fibril growth, will be useful to invent a potent inhibitor preventing the amyloid fibril growth of Abeta(1-40) and other proteins. PMID- 15533444 TI - Symmetric primary and tertiary structure mutations within a symmetric superfold: a solution, not a constraint, to achieve a foldable polypeptide. AB - In previous studies designed to increase the primary structure symmetry within the hydrophobic core of human acidic fibroblast growth factor (FGF-1) a combination of five mutations were accommodated, resulting in structure, stability and folding kinetic properties similar to wild-type (despite the symmetric constraint upon the set of core residues). A sixth mutation in the core, involving a highly conserved Met residue at position 67, appeared intolerant to substitution. Structural analysis suggested that the local packing environment of position 67 involved two regions of apparent insertions that distorted the tertiary structure symmetry inherent in the beta-trefoil architecture. It was postulated that a symmetric constraint upon the primary structure within the core could only be achieved after these insertions had been deleted (concomitantly increasing the tertiary structure symmetry). The deletion of these insertions is now shown to permit mutation of position 67, thereby increasing the primary structure symmetry relationship within the core. Furthermore, despite the imposed symmetric constraint upon both the primary and tertiary structure, the resulting mutant form of FGF-1 is substantially more stable. The apparent inserted regions are shown to be associated with heparin binding functionality; however, despite a marked reduction in heparin-binding affinity the mutant form of FGF-1 is surprisingly approximately 70 times more potent in 3T3 fibroblast mitogenic assays. The results support the hypothesis that primary structure symmetry within a symmetric protein superfold represents a possible solution, rather than a constraint, to achieving a foldable polypeptide. PMID- 15533445 TI - Protein-protein interactions: hot spots and structurally conserved residues often locate in complemented pockets that pre-organized in the unbound states: implications for docking. AB - Energetic hot spots account for a significant portion of the total binding free energy and correlate with structurally conserved interface residues. Here, we map experimentally determined hot spots and structurally conserved residues to investigate their geometrical organization. Unfilled pockets are pockets that remain unfilled after protein-protein complexation, while complemented pockets are pockets that disappear upon binding, representing tightly fit regions. We find that structurally conserved residues and energetic hot spots are strongly favored to be located in complemented pockets, and are disfavored in unfilled pockets. For the three available protein-protein complexes with complemented pockets where both members of the complex were alanine-scanned, 62% of all hot spots (DeltaDeltaG>2kcal/mol) are within these pockets, and 60% of the residues in the complemented pockets are hot spots. 93% of all red-hot residues (DeltaDeltaG>/=4kcal/mol) either protrude into or are located in complemented pockets. The occurrence of hot spots and conserved residues in complemented pockets highlights the role of local tight packing in protein associations, and rationalizes their energetic contribution and conservation. Complemented pockets and their corresponding protruding residues emerge among the most important geometric features in protein-protein interactions. By screening the solvent, this organization shields backbone hydrogen bonds and charge-charge interactions. Complemented pockets often pre-exist binding. For 18 protein-protein complexes with complemented pockets whose unbound structures are available, in 16 the pockets are identified to pre-exist in the unbound structures. The root-mean squared deviations of the atoms lining the pockets between the bound and unbound states is as small as 0.9A, suggesting that such pockets constitute features of the populated native state that may be used in docking. PMID- 15533446 TI - The optimal fraction of hydrophobic residues required to ensure protein collapse. AB - The hydrophobic interaction is the main driving force for protein folding. Here, we address the question of what is the optimal fraction, f of hydrophobic (H) residues required to ensure protein collapse. For very small f (say f<0.1), the protein chain is expected to behave as a random coil, where the H residues are "wrapped" locally by polar (P) residues. However, for large enough f this local coverage cannot be achieved and the thermodynamic alternative to avoid contact with water is burying the H residues in the interior of a compact chain structure. The interior also contains P residues that are known to be clustered to optimize their electrostatic interactions. This means that the H residues are clustered as well, i.e. they effectively attract each other like the H-monomers in Dill's HP lattice model. Previously, we asked the question: assuming that the H monomers in the HP model are distributed randomly along the chain, what fraction of them is required to ensure a compact ground state? We claimed there that f approximately p(c), where p(c) is the site percolation threshold of the lattice (in a percolation experiment, each site of an initially empty lattice is visited and a particle is placed there with a probability p. The interest is in the critical (minimal) value, p(c), for which percolation occurs, i.e. a cluster connecting the opposite sides of the lattice is created). Due to the above correspondence between the HP model and real proteins (and assuming that the H residues are distributed at random) we suggest that the experimental f should lead to percolating clusters of H residues over the highly dense protein core, i.e. clusters of the core size. To check this theory, we treat a simplified model consisting of H and P residues represented by their alpha-carbon atoms only. The structure is defined by the C(alpha)-C(alpha) virtual bond lengths, angles and dihedral angles, and the X-ray structure is best-fitted onto a face-centered cubic lattice. Percolation experiments are carried out for 103 single-chain proteins using six different hydrophobic sets of residues. Indeed, on average, percolating clusters are generated, which supports our theory; however, some sets lead to a better core coverage than others. We also calculate the largest actual hydrophobic cluster of each protein and show that, on average, these clusters span the core, again in accord with our theory. We discuss the effect of protein size, deviations from the average picture, and implications of this study for defining reliable simplified models of proteins. PMID- 15533447 TI - Independent ATPase activity of Hsp90 subunits creates a flexible assembly platform. AB - The ATPase activity of the molecular chaperone Hsp90 is essential for its function in the assembly of client proteins. To understand the mechanism of human Hsp90, we have carried out a detailed kinetic analysis of ATP binding, hydrolysis and product release. ATP binds rapidly in a two-step process involving the formation of a diffusion-collision complex followed by a conformational change. The rate-determining step was shown to be ATP hydrolysis and not subsequent ADP dissociation. There was no evidence from any of the biophysical measurements for cooperativity in either nucleotide binding or hydrolysis for the dimeric protein. A monomeric fragment, lacking the C-terminal dimerisation domain, showed no dependence on protein concentration and, therefore, subunit association for activity. The thermodynamic linkage between client protein binding and nucleotide affinity revealed ATP bound Hsp90 has a higher affinity for client proteins than the ADP bound form. The kinetics are consistent with independent Michaelis-Menten catalysis in each subunit of the Hsp90 dimer. We propose that Hsp90 functions in an open-ring configuration for client protein activation. PMID- 15533448 TI - Nucleic acid and prion protein interaction produces spherical amyloids which can function in vivo as coats of spongiform encephalopathy agent. AB - The infectious agent of transmissible spongiform encephalopathies (TSE) has been considered to be PrP(SC), a structural isoform of cellular prion protein PrP(C). PrP(SC) can exist as oligomers and/or as amyloid polymers. Nucleic acids induce structural conversion of recombinant prion protein PrP and PrP(C) to PrP(SC) form in solution and in vitro. Here, we report that nucleic acids, by interacting with PrP in solution, produce amyloid fibril and fibres of different morphologies, similar to those identified in the diseased brains. In addition, the same interaction produces polymer lattices and spherical amyloids of different dimensions (15-150 nm in diameters). The polymer lattices show apparent morphological similarity to the two-dimensional amyloid crystals obtained from linear amyloids isolated in vivo. The spherical amyloids structurally resemble "spherical particles" observed in natural spongiform encephalopathy (SE) and in scrapie-infected brains (TSE). We suggest that spherical amyloids, PrP(SC) amylospheroids, are probable constituents of the coat of the spherical particles found in vivo and the latter can act as protective coats of the SE and TSE agents in vivo. PMID- 15533449 TI - The structural transition of the prion protein into its pathogenic conformation is induced by unmasking hydrophobic sites. AB - A series of structural intermediates in the putative pathway from the cellular prion protein PrP(C) to the pathogenic form PrP(Sc) was established by systematic variation of low concentrations (<0.1%) of the detergent sodium dodecyl sulfate (SDS) or by the interaction with the bacterial chaperonin GroEL. Most extended studies were carried out with recombinant PrP (90-231) corresponding to the amino acid sequence of hamster prions PrP 27-30. Similar results were obtained with full-length recombinant PrP, hamster PrP 27-30 and PrP(C) isolated from transgenic, non-infected CHO cells. Varying the incubation conditions, i.e. the concentration of SDS, the GroEL and GroEL/ES, but always at neutral pH and room temperature, different conformations could be established. The conformations were characterized with respect to secondary structure as determined by CD spectroscopy and to molecular mass, as determined by fluorescence correlation spectroscopy and analytical ultracentrifugation: alpha-helical monomers, soluble alpha-helical dimers, soluble but beta-structured oligomers of a minimal size of 12-14 PrP molecules, and insoluble multimers were observed. A high activation barrier was found between the alpha-helical dimers and beta-structured oligomers. The numbers of SDS-molecules bound to PrP in different conformations were determined: Partially denatured, alpha-helical monomers bind 31 SDS molecules per PrP molecule, alpha-helical dimers 21, beta-structured oligomers 19-20, and beta structured multimers show very strong binding of five SDS molecules per PrP molecule. Binding of only five molecules of SDS per molecule of PrP leads to fast formation of beta-structures followed by irreversible aggregation. It is discussed that strongest binding of SDS has an effect identical with or similar to the interaction with GroEL thereby inducing identical or very similar transitions. The interaction with GroEL/ES stabilizes the soluble, alpha-helical conformation. The structure and their stabilities and particularly the induction of transitions by interaction of hydrophobic sites of PrP are discussed in respect to their biological relevance. PMID- 15533450 TI - Hetero-assembly between all-L- and all-D-amino acid transmembrane domains: forces involved and implication for inactivation of membrane proteins. AB - Protein-protein interactions within the membrane, partially or fully mediated by transmembrane (TM) domains, are involved in many vital cellular processes. Since the unique feature of the membrane environment enables protein-protein assembly that otherwise is not energetically favored in solution, the structural restrictions involved in the assembly of soluble proteins are not necessarily valid for the assembly of TM domains. Here we used the N-terminal TM domain (Tar 1) of the Escherichia coli aspartate receptor as a model system for examining the stereospecificity of TM-TM interactions in vitro and in vivo in isolated systems, and in the context of the full receptor. For this propose, we synthesized Tar-1 all-l and all-d amino acid TM peptides, a mutant TM peptide and an unrelated TM peptide. The data revealed: (i) Tar-1 all-d specifically associated with Tar-1 all-l within a model lipid membrane, as determined by using fluorescence energy transfer experiments; (ii) Tar-1 all-l and all-d, but not the control peptides, demonstrated a dose-dependant dominant negative effect on the Tar-1 TM homodimerization in the bacterial ToxR assembly system, suggesting a wild-type like interaction; and most interestingly, (iii) both Tar-1 all-l and all-d showed a remarkable ability to inhibit the chemotaxis response of the full-length receptor, in vivo. Peptide binding to the bacteria was confirmed through confocal imaging, and Western blotting confirmed that ToxR Tar-1 chimera protein levels are not affected by the presence of the exogenous peptides. These findings present the first evidence that an all-d TM domain peptide acts in vivo similarly to its parental all-l peptide and suggest that the dimerization of the TM domains is mainly mediated by side-chain interactions, rather than geometrically fitted conformations. In addition, the study provides a new approach for modifying the function of membrane proteins by proteolysis-free peptides. PMID- 15533451 TI - Comparative structural and energetic analysis of WW domain-peptide interactions. AB - WW domains are small globular protein interaction modules found in a wide spectrum of proteins. They recognize their target proteins by binding specifically to short linear peptide motifs that are often proline-rich. To infer the determinants of the ligand binding propensities of WW domains, we analyzed 42 WW domains. We built models of the 3D structures of the WW domains and their peptide complexes by comparative modeling supplemented with experimental data from peptide library screens. The models provide new insights into the orientation and position of the peptide in structures of WW domain-peptide complexes that have not yet been determined experimentally. From a protein interaction property similarity analysis (PIPSA) of the WW domain structures, we show that electrostatic potential is a distinguishing feature of WW domains and we propose a structure-based classification of WW domains that expands the existent ligand-based classification scheme. Application of the comparative molecular field analysis (CoMFA), GRID/GOLPE and comparative binding energy (COMBINE) analysis methods permitted the derivation of quantitative structure activity relationships (QSARs) that aid in identifying the specificity determining residues within WW domains and their ligand-recognition motifs. Using these QSARs, a new group-specific sequence feature of WW domains that target arginine-containing peptides was identified. Finally, the QSAR models were applied to the design of a peptide to bind with greater affinity than the known binding peptide sequences of the yRSP5-1 WW domain. The prediction was verified experimentally, providing validation of the QSAR models and demonstrating the possibility of rationally improving peptide affinity for WW domains. The QSAR models may also be applied to the prediction of the specificity of WW domains with uncharacterized ligand-binding properties. PMID- 15533452 TI - Cytochemical location of urease in the cell wall of two different lichen phycobionts. AB - The enzyme urease has been located in the cell wall of recently isolated phycobionts from Evernia prunastri and Xanthoria parietina lichens. Cytochemical detection is achieved by producing a black, electron-dense precipitate of cobalt sulfide proceeding from CO(2) evolved from urea in the presence of cobalt chloride. Cellular fractionation reveals that about 80% of total urease activity was associated to the cell wall on both phycobionts whereas only 20% was recovered as soluble protein. PMID- 15533453 TI - Calcium localisation by X-ray microanalysis and fluorescence microscopy in larvae of zooxanthellate and azooxanthellate corals. AB - X-ray microanalysis and fluorescence microscopy (Calcium Orangetrade mark) was used to determine the distribution of intracellular calcium (I(Ca)), in the form of total and ionic calcium respectively, in planulae and settled larvae of a zooxanthellate coral. The distribution of total calcium only was determined in larvae of an azooxanthellate coral. In azooxanthellate planulae and settled larvae, total I(Ca) concentration in the oral ectoderm was high and similar to that in seawater (SW). Calcium concentration did not vary (P > 0.05) between planulae and settled larvae. However, settled larvae accumulated large amounts of calcium in gastrodermal lipid-containing cells. In contrast, zooxanthellate planulae possessed significantly (P < 0.01) lower concentrations of total I(Ca) within ectodermal cells in comparison to settled larvae. In addition, in settled zooxanthellate larvae total calcium concentration in the mesogloea and coelenteron was significantly (P < 0.05) higher than in the oral ectodermal and gastrodermal cells, respectively. Total I(Ca) concentrations in the oral ectoderm of settled larvae were also significantly (P < 0.01) lower than that of the calicoblastic ectoderm. In zooxanthellate settled larvae, ionic I(Ca) levels in the aboral epithelium surrounding rapidly growing septa were high. These levels increased significantly (P < 0.05) within the tissue surrounding growing septa after incubation in high-calcium SW. PMID- 15533454 TI - Distribution of alpha7 and alpha4 nicotinic acetylcholine receptor subunits in several tissues of Triturus carnifex (Amphibia, Urodela). AB - The distribution of neuronal and non-neuronal mRNAs for alpha7 and alpha4 nicotinic acetylcholine receptor subunits was investigated in Triturus carnifex tissues using the in situ hybridization approach. The findings reveal a composite pattern of expression only partially overlapping for the two subunits; subunit alpha7 seems to be expressed widely throughout nervous, gastrointestinal and skin tissues, while alpha4 is present in a restricted number of cells of nervous and gastrointestinal tissue. We also found a specific pattern for each subunit; alpha7 and alpha4 associated exclusively to the epidermal glands and hypophysis, respectively; this is probably due to alternative roles that nicotinic acetylcholine receptors play in regulating physiological functions of non neuronal amphibian tissues, rather than as mere neurotransmitters in the nervous system. PMID- 15533455 TI - The structure and ultrastructure of the sinus venosus in the mature dogfish (Scyliorhinus canicula): the endocardium, the epicardium and the subepicardial space. AB - The sinus venosus of fish is the most caudal chamber of the heart. It is often reduced in teleosts but well developed in elasmobranchs. The sinus venosus of the dogfish (Scyliorhinus canicula) is vital, since it harbours key elements such as a little known neuroendocrine system and the nodal tissue. However, the study of its structure is still incomplete. We examined the endocardium, epicardium and subepicardium of the sinus venosus in mature dogfishes. The wall is 100-250 microm thick and comprises three main layers. Large bundles of myocardial cells occupy the middle layer. The endothelial ensheathing is composed of thin endocardial cells with prominent nuclei towards the lumen, whose cytoplasm contains numerous dense bodies and moderately dense bodies, 150-800 nm in diameter and large vacuoles. The possible functions of these organelles are discussed. The outermost layer is made of a robust sheet of cuboidal epicardial cells separated from the subepicardium by a conspicuous basal lamina. Numerous microvilli towards the pericardial cavity and elliptical vesicles are located in the apex of epicardial cells. A thick layer richly endowed with dense bundles of collagen fibres forms the subepicardial space. This structure should be contrasted with the venous return mechanism of elasmobranchs. PMID- 15533456 TI - Morphological study of the parathyroid gland and thyroid C cell in senescence accelerated mouse (SAMP6), a murine model for senile osteoporosis. AB - SAMP6, a substrain of senescence-accelerated mouse, was developed as an animal model for senile osteoporosis. We investigated the morphology of the parathyroid gland and thyroid C cell, together with the serum parathyroid hormone (PTH) and calcitonin (CT) in SAMP6 and age-matched normal mice SAMR1. We did not find any significant differences between SAMR1 and SAMP6 at 1 month of age with regard to the serum PTH level and the morphology of the parathyroid glands. As compared with SAMR1, the serum PTH level was significantly higher in SAMP6 at 2, 5 and 12 months of age. In the parathyroid chief cells of SAMP6 at 2, 5 and 12 months of age, the Golgi complexes and the cisternae of the granular endoplasmic reticulum were well developed. Numerous secretory granules were located near the plasma membranes and mitoses were sometimes observed. There was no marked difference between SAMR1 and SAMP6 regarding the morphology of the thyroid C cells and the serum CT level. These findings suggest that the secretory activity of the parathyroid gland is stimulated in SAMP6 at 2, 5 and 12 months of age. The parathyroid follicle was sometimes found in SAMP6, and the significance of this structure was also discussed. PMID- 15533457 TI - Influence of hormonal replacement on the ventral lobe of the prostate of rats (Rattus norvegicus albinus) submitted to chronic ethanol treatment. AB - The harmful influence of the chronic alcohol ingestion on the male reproductive system leads to important alterations including hypogonadism and feminization, besides the morphological and functional disorganization of the different sexual glands. So, the aim of this study was to analyse the structural changes on the ventral lobe of the prostate of rats with hormonal replacement associated to chronic alcohol ingestion. A total of 30 rats (Rattus norvegicus albinus) was divided into three groups: control-received water; alcoholic-received ethanol diluted to 20% and hormone-treated alcoholic-received ethanol diluted to 20% associated with the administering of testosterone (5mg/kg of weight) during the last 30 days of treatment. After 150 days of treatment, the animals were sacrificed, the prostate removed and submitted to transmission and scanning electron microscopies, histochemical analysis for acid phosphatase, testosterone level and stereologic analysis. In the alcoholic group the results demonstrated reduction of the total cellular volume and disorganization of the organelles involved in the secretory process. It was characterized a partial recovery of the cellular volume after treatment with testosterone. It was concluded that the ethanol impaired the cellular morphology and the hormonal replacement by itself did not bring about efficient remodeling of the organelles responsible for the secretory process. PMID- 15533458 TI - Hyaluronic acid and autologous synovial fluid induce chondrogenic differentiation of equine mesenchymal stem cells: a preliminary study. AB - Mesenchymal stem cells (MSC) have the potential to differentiate into distinct mesenchymal tissues including cartilage, which suggest these cells as an attractive cell source for cartilage tissue engineering approaches. Our objective was to study the effects of TGF-beta1, hyaluronic acid and synovial fluid on chondrogenic differentiation of equine MSC. For that, bone marrow was aspirated from the tibia of one 18-month-old horse (Haflinger) and MSC were isolated using percoll-density centrifugation. To promote chondrogenesis, MSC were centrifuged to form a micromass and were cultured in a medium containing 10 ng/ml TGF-beta1 or 0.1mg/ml hyaluronic acid (Hylartil, Ostenil) or either 5%, 10% or 50% autologous synovial fluid as the chondrogenesis inducing factor. Differentiation along the chondrogenic lineage was documented by type II collagen and proteoglycan expression. MSC induced by TGF-beta1 alone showed the highest proteoglycan expression. Combining TGF-beta1 with hyaluronic acid could not increase the proteoglycan expression. Cultures stimulated by autologous synovial fluid (independent of concentration) and hyaluronic acid demonstrated a pronounced, but lower proteoglycan expression than cultures stimulated by TGF beta1. The expression of cartilage-specific type II collagen was high and about the same in all stimulated cultures. In summary, hyaluronic acid and autologous synovial fluid induces chondrogenesis of equine mesenchymal stem cells, which encourage tissue engineering applications of MSC in chondral defects, as the natural environment in the joint is favorable for chondrogenic differentiation. PMID- 15533459 TI - Stages and duration of spermatogenesis in the domestic ferret (Mustela putorius furo). AB - Classification of seminiferous tubules is the basis for understanding normal and abnormal spermatogenesis. The aim of the present study was to determine spermatogenic stages and the duration of the cycle in the domestic ferret using bromodeoxyuridine (BrdU) as a tracer. Eleven adult male ferrets that were maintained in a breeding condition were used. Testicular sections were stained with the periodic acid-Schiff reaction for light microscopy. To determine the cycle duration, six ferrets were injected intraperitoneally with BrdU, and testes were collected 3h later and 10 days and 3h later. BrdU was detected by immunohistochemistry. Seminiferous tubules were classified into eight stages, and frequencies of stages I-VIII were 10.6, 2.2, 7.9, 13.1, 22.3, 21.9, 14.0 and 8.0%, respectively. The most advanced BrdU-labeled cells at 3h post-injection were leptotene spermatocytes in stage VI and those at 10 days and 3h were pachytene spermatocytes in stage V. From differences in stage frequency and BrdU staining frequency between two time points, the duration of one cycle was estimated to be 13.0 days. The present observations indicate that stages and the cycle duration of the ferret spermatogenesis are similar to those reported in other carnivores. PMID- 15533460 TI - The ultrastructure of the peculiar synspermia of some Dysderidae (Araneae, Arachnida). AB - The present study reports on the ultrastructure features of spermatozoa and spermatogenesis of several species of Dysderidae (Dysdera crocata, Dysdera erythrina, Dysdera ninnii, Harpactea arguta, Harpactea piligera, Dasumia taeniifera). Dysderid spiders are known to possess a peculiar sperm transfer form known as synspermia, characterized by fused spermatozoa surrounded by a secreted sheath. Until now the exact mode of formation of the synspermia is unknown. The present study demonstrates that the spermatids are connected via narrow cell bridges during the entire spermiogenesis as is usual, although in Dysderidae they do not separate at end of the spermiogenesis. Instead, they fuse completely within the testes shortly after the spermatid has coiled to get a spherical shape. The number of fusing sperm cells is different in the different observed species. The species of the genus Harpactea thus have synspermia consisting of two fused spermatozoa; whereas in the species of the genus Dysdera four sperm cells are fused and in D. taeniifera at least three spermatozoa are fused. In contrast with other known families with this peculiar form transfer of sperm, the synspermia in Dysderidae are mainly characterized by a conspicuous vesicular area which extends through the entire synspermium surrounding the cell organelles. Thus, all main cell components (e.g., nucleus, acrosomal vacuole, and axoneme) are covered by the vesicular membrane. The vesicular area seems to be functional and probably it is important during sperm activation in female genital system. Simultaneously to the extension of the vesicular area, the synspermium accumulates large amounts of glycogen. The glycogen is mainly located around the centriolar adjunct and along the axoneme accompanying the postcentriolar elongation of the nucleus. A further peculiar feature is the extremely elongated acrosomal vacuole, which seems to be synapomorphic trait for sperm cells of dysderids. Interestingly, spermatogenesis, including the fusion, exclusively occurs within the testes (in contrast to the formation of coenospermia). In the vas deferens only synspermia were found. The secreted sheath surrounding the spermatozoa is finally synthesized in the parts of the vasa deferentia, which are close to the genital opening where numerous vacuoles and microvilli are seen in the epithelial cells. PMID- 15533461 TI - Use of bone anchors in surgery for stress urinary incontinence: pro. PMID- 15533462 TI - Bone anchors for stress urinary incontinence in 2004: con. PMID- 15533463 TI - Combining external beam radiotherapy with prostate brachytherapy: issues and rationale. PMID- 15533464 TI - Modified Thiele massage as therapeutic intervention for female patients with interstitial cystitis and high-tone pelvic floor dysfunction. AB - OBJECTIVES: To evaluate the effectiveness of transvaginal manual therapy of the pelvic floor musculature (Thiele massage) in symptomatic female patients with interstitial cystitis and high-tone dysfunction of the pelvic floor. METHODS: A total of 21 women with documented interstitial cystitis and high-tone pelvic floor dysfunction underwent transvaginal massage using the Thiele technique twice a week for 5 weeks. Symptoms were evaluated before massage, at protocol conclusion, and at a mean of 4.5 months after therapy completion (long-term follow-up). The response to treatment was evaluated through the O'Leary-Sant Interstitial Cystitis Symptom and Problem Indexes, Likert Visual Analogue Scales for urgency and pain, and Short-Form 12-item (SF-12) Quality-of-Life Scale, and through changes in the physical examination findings using a 5-point modified Oxford Scale to document pelvic floor tenderness. RESULTS: A statistically significant improvement was seen in the Symptom and Problem Indexes of the O'Leary-Sant Questionnaire (P = 0.015 and P = 0.039, respectively), Likert Visual Analogue Scales for urgency and pain (P = 0.001 and P = 0.005, respectively), the Physical and Mental Component Summary from the SF-12 Quality-of-Life Scale (P = 0.049 and P = 0.044, respectively), and the modified Oxford Scale (P <0.05) after protocol completion. At long-term follow-up, the O'Leary-Sant Interstitial Cystitis Symptom and Problem Indexes (P = 0.049 and P = 0.02, respectively), Likert Visual Analogue Scales for urgency and pain (P = 0.004 and P = 0.005, respectively), and modified Oxford Scale for three of four muscles in the pelvic floor (P <0.05) remained significantly improved. CONCLUSIONS: Thiele massage appears to be very helpful in improving irritative bladder symptoms in patients with interstitial cystitis and high-tone pelvic floor dysfunction in addition to decreasing pelvic floor muscle tone. PMID- 15533465 TI - Prevalence of interstitial cystitis in young women. AB - OBJECTIVES: Traditional epidemiologic studies have significantly underestimated interstitial cystitis (IC) prevalence because they surveyed populations for diagnosed cases rather than screening for IC symptoms and evaluating suspected cases. Our earlier data have suggested that IC affects almost 25% of women. To test this hypothesis, we used a validated IC symptom questionnaire and intravesical potassium sensitivity testing (PST), history, and physical examination to determine the prevalence of IC in a fixed population of young women. METHODS: All female members of the University of California, San Diego, third-year medical student class were asked to complete the Pelvic Pain and Urgency/Frequency (PUF) scale. All those scoring 7 or greater were asked to undergo clinical evaluation, including urinalysis, urine culture, and PST. RESULTS: Of 52 potential subjects, 49 (median age 26 years) completed the PUF scale. Of the 49, 15 (30.6%) scored 7 or greater; 5 of those 15 volunteered for PST. All 5 had negative urinalysis findings and were PST positive, for a 10% (5 of 52) rate of positive voluntary PST in the medical student population. All 15 subjects with PUF scores of 7 or greater reported being sexually active. Dyspareunia was present in 13 (87%) of the 15 women, including all 5 PST-positive subjects. CONCLUSIONS: We identified probable IC in 30.6% and documented IC in a minimum of 10% of the female medical students. These data suggest the estimate of IC prevalence in the United States should be vastly increased from approximately 1.5 million to perhaps 25 to 30 million women and that IC is highly prevalent in young women. Screening for IC-specific symptoms is a useful method for identifying undiagnosed IC cases. PMID- 15533466 TI - Botulinum toxin a has antinociceptive effects in treating interstitial cystitis. AB - OBJECTIVES: To present clinical evidence with botulinum toxin A (BTX-A) suggesting an antinociceptive role in patients with interstitial cystitis (IC). Intriguing evidence in a somatic pain model has suggested that BTX-A injection may have an antinociceptive effect on both acute and chronic (inflammatory) pain. METHODS: Thirteen female patients (6 in the United States and 7 in Poland) with IC according to the criteria of the National Institute of Diabetes, Digestive and Kidney Disease were included. Under short general anesthesia or sedation, 100 to 200 U of Dysport (Polish patients) or Botox (U.S. patients) was injected through a cystoscope into 20 to 30 sites submucosally in the trigone and floor of the bladder. Patients were evaluated with the O'Leary-Sant validated IC questionnaire or with voiding charts and a visual analog pain scale 1 month postoperatively and at subsequent 3-month intervals. The Polish patients also underwent pretreatment and post-treatment urodynamic evaluations. RESULTS: Overall, 9 (69%) of 13 patients noted subjective improvement after BTX-A treatment. The Interstitial Cystitis Symptom Index and Interstitial Cystitis Problem Index mean scores improved by 71% and 69%, respectively (P <0.05). Daytime frequency, nocturia, and pain by visual analog scale decreased by 44%, 45%, and 79%, respectively (P <0.01). The first desire to void and maximal cystometric capacity increased by 58% and 57%, respectively (P <0.01). CONCLUSIONS: Our results suggest that BTX-A has an antinociceptive effect on bladder afferent pathways in patients with IC, producing both symptomatic and functional (ie, urodynamic) improvements. PMID- 15533468 TI - Self-report of prostatitis and its risk factors in a random sample of middle-aged men. AB - OBJECTIVES: To determine the risk factors for the self-reported history of prostatitis and to determine whether a self-reported history of prostatitis is related to the diagnosis of prostate cancer. METHODS: Random digit dialing was used to select a group of controls aged 40 to 64 years without prostate cancer matched by age with a group of patients with prostate cancer in a study on the epidemiology of prostate cancer. Controls were divided into those who reported a diagnosis of prostatitis (cases) and those who denied ever having had prostatitis (controls). We adjusted for the time from a prostatitis diagnosis to the in person interview. We also compared the number of men with a prostatitis diagnosis to the number of men diagnosed and not diagnosed with prostate cancer. RESULTS: Of the 645 control men without a history of prostate cancer, 58 (9.0%) reported a diagnosis of prostatitis. The men with prostatitis had a mean age of 39.5 years at diagnosis. The urinary symptoms among prostatitis cases and controls was similar. Prostatitis cases more frequently reported urinary (P < or =0.05) or urethral infections (P < or =0.01) before diagnosis. Men with prostatitis were more likely to have procedures aimed at diagnosing both prostatic and other diseases. Men with prostate cancer reported a diagnosis of prostatitis more often than the noncancer controls (13.6% versus 9.0%). After controlling for the number of prostate-specific antigen tests this difference disappeared. CONCLUSIONS: Nine percent of a randomly selected group of middle-aged men reported they had been diagnosed with prostatitis. The prevalence of a prostatitis diagnosis is similar in men with and without prostate cancer. PMID- 15533470 TI - Initial, long-term, and durable responses to terazosin, placebo, or other therapies for chronic prostatitis/chronic pelvic pain syndrome. AB - OBJECTIVES: To evaluate the initial, long-term, and durable response rates to terazosin, placebo, or other therapies in patients with chronic prostatitis/chronic pelvic pain syndrome. METHODS: A total of 100 subjects, aged 20 to 50 years, who met the National Institutes of Health criteria for chronic prostatitis/chronic pelvic pain syndrome and had not previously been treated with alpha-blockers, were entered in a 14-week, double-blind comparison of terazosin or placebo therapy. Nonresponders and responders with subsequent relapse were treated with terazosin or other medications (open label). The criterion for response was a score of 0 to 2 on the National Institutes of Health Chronic Prostatitis Symptom Index quality-of-life item. The initial response was evaluated at week 14, and the long-term response was evaluated after a median of 38 weeks (range 34 to 42), regardless of any additional treatment. A durable response was defined as an initial response without additional treatment. RESULTS: Of the 43 patients in the terazosin group, 24 (56%) had an initial response compared with 14 (33%) of 43 subjects in the placebo group (P = 0.03). Long-term responses were noted in 23 (56%) of 41 assessable subjects treated with terazosin initially compared with 12 (32%) of 38 assessable subjects treated with placebo (P = 0.03). Of the nonresponders and initial responders with relapse, 7 (41%) of 17 subjects responded to terazosin compared with 7 (21%) of 34 given other treatment (P = 0.12). Durable responses occurred in 18 (44%) of the 41 assessable patients treated initially with terazosin and in 6 (16%) of 38 treated initially with placebo (P = 0.01). CONCLUSIONS: Patients treated with terazosin were more likely to have initial, long-term, and durable responses than those treated with placebo. PMID- 15533471 TI - Novel algorithm for improving accuracy of ultrasound measurement of residual urine volume according to bladder shape. AB - OBJECTIVES: To suggest a novel algorithm based on individually determined proportionality constants to improve the accuracy of noninvasively calculated residual urine volume using ultrasound bladder images. METHODS: The residual urine volume of 116 subjects was calculated from ultrasound images obtained using a conventional two-dimensional ultrasound device at two institutions. The proposed method uses individually determined proportionality constants for specific bladder shapes. The corresponding true residual volumes were determined by measuring the catheterized volume. The accuracy of our method was confirmed by comparing the mean and standard deviation of the fractional absolute error (FAE) versus true volume with values obtained using other methods, which rely on fixed constants. RESULTS: The calculated residual bladder volume of 116 patients correlated significantly (r = 0.94, P <0.001) and the differences from the true values were not statistically significant (P = 0.42). The mean FAE was 0.17 +/- 0.10. Subgroups divided by sex, postvoid residual urine volume (over and under 150 mL), and institution produced different results (mean FAE for men 0.15 +/- 0.11 and for women 0.18 +/- 0.12; mean FAE for greater than 150 mL, 0.16 +/- 0.11 and for less than 150 mL, 0.17 +/- 0.14; mean FAE for Seoul National University Hospital 0.15 +/- 0.10 and for Seoul National University Bundang Hospital 0.18 +/ 0.18). Our method provided better accuracy than other known formulas using a conventional two-dimensional ultrasound scanner. The latter multiply the product of the sagittal height and depth and transverse width by a fixed coefficient. CONCLUSIONS: Our method, based on individually determined proportionality constants, improved the accuracy of residual bladder volume determinations. The proposed algorithm can easily be implemented on a personal computer connected to a conventional ultrasound imaging unit. PMID- 15533472 TI - A novel dual-diameter ureteroscope working channel: impact on irrigant flow. AB - OBJECTIVES: To compare irrigant flow characteristics through standard working channels (3.6F to 4.5F) and a dual-diameter working channel. Irrigant flow is critical for adequate visualization during endoscopic procedures. METHODS: Working channels were created out of 80 cm of light wall polytetrafluoroethylene (PTFE) tubing (inner diameter 3.6F, 4.0F, and 4.5F) with a male Luer connector attached to one end by epoxy resin. The dual-diameter working channel was created with a 70-cm segment of PTFE (4.5F inner diameter, 0.059 in.) and a 10-cm segment of PTFE (3.6F inner diameter, 0.047 in.) with a male Luer connector attached to the free end of the 4.5F tubing. Stone basket shafts (100 cm in length, outer diameter 1.9F, 2.4F, and 3.0F) were created out of unmodified polyimide tubing with a 0.018-in.-diameter nitinol mandrel epoxy core for stability. Irrigant flow was measured at 100 mm Hg pressure for 1 minute with an empty channel and with stone basket shafts in the channel. RESULTS: The flow rates were significantly greater with the dual-diameter working channel than with the standard flexible ureteroscope (3.6F) working channel using an empty channel (79.2 versus 44.1 mL/min, P = 0.0001), 1.9F basket (35.9 versus 10.0 mL/min, P = 0.003), 2.4F basket (20.7 versus 4.3 mL/min, P = 0.002), and 3.0F basket (6.0 versus 0.7 mL/min, P = 0.0002) sheath. CONCLUSIONS: A dual-diameter working channel may optimize irrigant flow characteristics for flexible ureteroscopes while maintaining a small distal tip diameter. PMID- 15533473 TI - Percutaneous nephrostomy versus indwelling ureteral stents in the management of extrinsic ureteral obstruction in advanced malignancies: are there differences? AB - OBJECTIVES: To compare the complications and morbidities after placement of a percutaneous nephrostomy (PCN) tube or an internal ureteral stent (IUS) in the management of malignant ureteral obstruction in patients with advanced malignancy. METHODS: A retrospective analysis was performed on a total of 148 patients (80 women and 68 men, mean age 57.3 years, range 20 to 84) with malignant ureteral obstruction, who underwent PCN tube placement (n = 80) or IUS placement (n = 68) between January 2000 and December 2002. The incidence of fever and acute pyelonephritis was expressed as the number of episodes per 100 person days. RESULTS: The accumulated incidence of fever and acute pyelonephritis was not different in the two groups. The accumulated incidence and the incidence of febrile episodes in the IUS group was 10.3% and 0.0004/100 person-days; the corresponding values for the PCN group were 15.0% and 0.2154/100 person-days. The incidence of acute pyelonephritis in the IUS and PCN groups was 0.0002/100 person days and 0.0005/100 person-days, respectively. These patients were treated conservatively and recovered uneventfully. The difference in overall stent related or catheter-related complications between the IUS and PCN groups was not statistically significant. The accumulated incidence of failed diversion due to obstruction was 11% (8 of 68) and 1.3% (1 of 80) in the IUS and PCN groups, respectively (P = 0.012). CONCLUSIONS: Our results have demonstrated that morbidities after internal or external diversion were minimal in cases of malignant obstruction. However, patients scheduled to receive an IUS should be more carefully monitored for ongoing obstruction than patients scheduled for PCN tube placement. PMID- 15533474 TI - Endourologic procedures combined with other surgical and urologic interventions- early experience. AB - OBJECTIVES: To assess the feasibility of performing endourologic interventions combined with other operations during the same operative session. METHODS: Eighteen patients underwent simultaneous operations endourologically for upper urinary tract pathologic findings and other surgical and urologic indications. The operating time, technical feasibility, operative success, complications, hospital stay, and patient satisfaction were analyzed. RESULTS: The average patient age was 63 years (range 40 to 83). Five patients underwent percutaneous nephrolithotomy combined with either contralateral laparoscopic nephrectomy, contralateral open nephrectomy, radical retropubic prostatectomy, inguinal/umbilical hernia repair, transurethral resection of prostate, or cystolithotripsy. Thirteen patients underwent 15 retrograde endoscopic procedures (13 for stone disease and 2 for diagnostic purposes) that were combined with open contralateral nephrectomy, inguinal hernia repair, circumcision, closure of ileostomy, transurethral resection of bladder tumor, excision of lymphoma of thigh, drainage and sclerozation of hydrocele, or percutaneous gastrostomy. All procedures were successfully completed without complications. The average hospital stay was 5 days (range 3 to 6) in the percutaneous nephrolithotomy group and 2 days (range 1 to 5) in the retrograde endoscopic procedure group. The duration of hospitalization was related to the more complex operation; combining the procedures did not prolong it. The average follow-up was 11 months (range 3 to 24). All patients were highly satisfied because they were spared the need for more than one surgical session. CONCLUSIONS: Our results support the concept of performing simultaneous endourologic procedures and other operations during one surgical session. This approach obviates the need for repeated anesthesia, patient inconvenience, the psychological stress related to multiple operations, and reduces the total hospital stay. PMID- 15533475 TI - Prediction of postoperative renal function by preoperative serum creatinine level and three-dimensional diagnostic image reconstruction in patients with renal cell carcinoma. AB - OBJECTIVES: To investigate retrospectively whether postoperative renal function in patients with renal cell carcinoma can be preoperatively predicted by a combination of the preoperative serum creatinine (sCr) and the renal parenchymal volume (RPV) estimated by three-dimensional image reconstruction of the preoperative diagnostic imaging. METHODS: Of 155 patients who had undergone radical nephrectomy, 76 were eligible for inclusion in our study (group 1). Group 2 was comprised of 26 of 37 patients who had undergone partial nephrectomy. The postoperative RPV in both groups was estimated from the preoperative computed tomography scans or magnetic resonance imaging and were compared with the actual RPV estimated from the postoperative imaging using a three-dimensional image reconstruction program. The postoperative creatinine clearance (Ccr) was predicted from the preoperative Ccr calculated from the sCr level and by the ratio of the postoperative/preoperative RPV. The correlations between the predicted postoperative Ccr and the actual, measured postoperative Ccr were analyzed statistically. RESULTS: In both groups, a statistically significant correlation was found between the postoperative RPV, estimated from the preoperative images and postoperative images. The predicted postoperative Ccr correlated significantly with the actual, measured postoperative Ccr in group 1 (r = 0.86, P <0.0001) and group 2 (r = 0.98, P <0.0001). The postoperatively increased sCr achieved stable levels within 2 to 4 weeks after nephrectomy and showed no statistically significant subsequent changes during 3 years of follow up. CONCLUSIONS: The present results demonstrated that prediction of the postoperative Ccr, using the preoperative sCr and the postoperative RPV estimated from the preoperative routine diagnostic imaging, is a simple and reliable method for the evaluation of early and medium-term postoperative renal function. PMID- 15533476 TI - Management of renal masses in patients medically unsuitable for nephrectomy- natural history, complications, and outcome. AB - OBJECTIVES: To examine a group of elderly patients with much larger tumors who were deemed unlikely to survive surgery or who would require dialysis postoperatively to establish the natural history of larger renal tumors if left untreated. Little is known of the growth rate and natural history of renal cancer progression because the tumor is usually removed in those patients who can tolerate surgery. The only published data have been in relation to unfit patients with small, homogeneous, well-circumscribed tumors less than 4 cm. METHODS: We identified 36 patients whose tumor had not been removed, without evidence of metastasis at diagnosis, from a database of 421 patients with renal cancer. The data were examined retrospectively for symptoms, survival, and size change. RESULTS: The mean age of the patients treated conservatively was 76.1 years (range 56 to 91), with median tumor size of 6.0 cm (range 3.5 to 20.0) at diagnosis. The median follow-up period was 24 months (range 3 to 136). Of the 36 patients, 13 had died at follow-up, 8 of an unrelated illness and 5 of an unknown cause with no radiologic evidence of progression but severe comorbidity. The median time to death was 9 months (range 3 to 24) after diagnosis. One patient developed metastasis at 132 months and was still alive at 136 months of follow up. Significant hematuria occurred in 11% of the patients and was successfully managed either conservatively or by embolization. The tumor size was unchanged in most patients during the follow-up period. CONCLUSIONS: In elderly patients, or those with severe comorbidity, conservative management of larger renal masses is a reasonable and safe option. PMID- 15533477 TI - Robotic-assisted laparoscopic partial nephrectomy: technique and initial clinical experience with DaVinci robotic system. AB - OBJECTIVES: To develop and assess the feasibility of laparoscopic partial nephrectomy performed using the daVinci robotic system. METHODS: Between November 2002 and August 2003, 13 patients with solid or suspicious cystic renal masses underwent robotic-assisted laparoscopic partial nephrectomy. In 8 cases, an intra arterial catheter was inserted for renal cooling before occlusion of the renal artery. The remaining 5 patients underwent partial nephrectomy after the renal hilum had been clamped. Tumor excision and intracorporeal suturing were performed entirely with telerobotics. The perioperative data and pathologic results were retrospectively reviewed. RESULTS: The mean lesion diameter was 3.5 cm (range 2.0 to 6.0). The mean operative time was 215 minutes (range 130 to 262), and the mean blood loss was 170 mL (range 50 to 300). The mean warm ischemia was 22 minutes (range 15 to 29), and the mean cold ischemia time was 33 minutes (range 18 to 43). The length of hospital stay averaged 4.3 days (range 2 to 7). The resected lesions included renal cell carcinoma in 10, oncocytoma in 2, and a complex renal cyst in 1. In 1 case, a positive margin occurred despite negative frozen sections; laparoscopic nephrectomy was performed and showed no residual tumor. One patient experienced postoperative ileus. At 2 to 11 months of follow-up, no recurrence had been observed. CONCLUSIONS: Robotic-assisted partial nephrectomy is feasible. Robotic partial nephrectomy can be safely performed using a transperitoneal or retroperitoneal approach. A second scrubbed assistant is mandatory to provide assistance using conventional laparoscopic instruments with this technique. PMID- 15533478 TI - Prospective, randomized controlled study: transperitoneal laparoscopic versus retroperitoneoscopic radical nephrectomy. AB - OBJECTIVES: To evaluate the transperitoneal and retroperitoneal approaches for endoscopic radical nephrectomy in a prospective randomized manner to assess the possible differences in the outcome related to patients' morbidity and technical difficulty for the surgeon. METHODS: A total of 40 patients with Stage cT1-T2 were randomized into two equal groups: laparoscopic radical nephrectomy (LRN) and retroperitoneoscopic radical nephrectomy (RRN). The patient demographics and tumor characteristics were comparable. Two surgeons with differing experience performed an equal number of procedures in both treatment arms. The outcome was compared, and the technical difficulty for the surgeon and assistant was assessed with the European scoring system. RESULTS: All procedures were completed without a need for conversion. No statistically significant differences were found between the two approaches in terms of the number and size of the trocars used, length of incision, specimen weight, pathologic stage, operative time, need for additional procedures such as adrenalectomy and/or lymph node sampling, estimated blood loss, need for blood transfusions, analgesic requirement, length of hospital stay, or the incidence of minor or major complications. All patients in the LRN group resumed oral intake on postoperative day 1, but only 75% did so in the RRN group. The technical difficulty score for either the surgeon or the assistant did not differ significantly between the two groups. Both approaches allowed complete tumor excision. The robotic assistance system (AESOP) was more difficult with RRN compared with LRN. CONCLUSIONS: This first prospective randomized study comparing LRN and RRN did not find any real difference between the two approaches in relation to patient morbidity or the technical difficulty for the surgeon. PMID- 15533479 TI - Laparoscopic-assisted nephrectomy with inferior vena cava tumor thrombectomy: preliminary results. AB - OBJECTIVES: To evaluate the feasibility and outcome of laparoscopic-assisted radical nephrectomy and inferior vena cava (IVC) tumor thrombectomy in patients with renal cell carcinoma and level I IVC tumor thrombus. METHODS: The clinical, operative, and pathologic data were retrospectively obtained from patients undergoing the above-mentioned procedure for renal tumors involving the IVC. This approach involved laparoscopic dissection of the kidney and renal vasculature/IVC. After renal artery ligation, an 8 to 12-cm incision was made from the tip of the 11th rib extending anteriorly toward the midline. Through this incision, a Satinsky vascular clamp was placed on the IVC in such a way as to include all the caval thrombus. The tumor thrombus was removed en bloc with the kidney and the cavotomy repaired with a running suture. RESULTS: Four obese patients underwent transperitoneal laparoscopic-assisted right nephrectomy with inferior vena cava (IVC) thrombectomy. The mean tumor size was 9 cm (range 6 to 13), with the thrombus extending 2 cm into the IVC in all cases. Patients had a mean body mass index of 32.8 (range 30.5 to 37.2) and a mean American Society of Anesthesiologists score of 2.8 (range 2 to 3). The mean operative time was 248 minutes (range 225 to 274). The mean estimated blood loss was 517 mL (range 250 to 900). No intraoperative or postoperative complications occurred. The mean hospital stay was 6.2 days (range 4 to 11, median 5). CONCLUSIONS: Laparoscopic assisted nephrectomy and IVC thrombectomy is a difficult but feasible procedure. This approach allows a smaller incision than a typical open approach. Additional studies are needed to examine the advantages of this approach over a pure open approach. PMID- 15533480 TI - Laparoscopic versus open cytoreductive nephrectomy for metastatic renal cell carcinoma. AB - OBJECTIVES: To compare the outcomes of those patients who underwent laparoscopic versus open cytoreductive nephrectomy. Cytoreductive nephrectomy before systemic therapy has been shown to offer a survival advantage compared with systemic therapy alone for metastatic renal cell carcinoma. METHODS: We reviewed the outcomes of all patients who underwent either open or laparoscopic cytoreductive nephrectomy between 2000 and 2003. The inclusion criteria included patients with tumors 15 cm or less without local invasion, venous involvement, or bulky local adenopathy who had concurrent metastatic disease. A total of 64 patients (22 in the laparoscopic group and 42 in the open group) fulfilled these criteria. The parameters measured were age, tumor size, operative time, estimated blood loss, complications, length of hospital stay, percentage of patients receiving systemic therapy, and the interval to the start of systemic therapy. Kaplan-Meier survival estimates were compared. RESULTS: Patients who underwent laparoscopic cytoreductive nephrectomy had a shorter length of stay (2.3 versus 6.1 days) and less operative blood loss (288 versus 1228 mL) than those who underwent open nephrectomy. Patients in the laparoscopic group received systemic therapy sooner after surgery (36 versus 61 days) than those in the open group. The Kaplan-Meier survival estimates were similar for both groups, with a 1-year survival rate of 61% in the laparoscopic group and 65% in the open group. CONCLUSIONS: With judicious patient selection, laparoscopic cytoreductive nephrectomy can be performed safely, with minimal morbidity, and may shorten the interval from nephrectomy to the start of systemic therapy. PMID- 15533481 TI - Laparoscopic radical cystectomy with continent urinary diversion (rectosigmoid pouch) performed completely intracorporeally: an intermediate functional and oncologic analysis. AB - OBJECTIVES: To present our experience with the first series of rectosigmoid pouch creation performed completely laparoscopically for continent urinary diversion after radical cystectomy to treat transitional cell carcinoma of the bladder. We evaluated the intermediate functional and oncologic outcomes. METHODS: Between April 2000 and January 2004, 20 patients underwent laparoscopic radical cystectomy with urinary diversion for transitional cell carcinoma at the Department of Urology, Charite Hospital, Campus Mitte. Of the 20 patients, 12 received a rectosigmoid pouch for urinary diversion. The procedures were performed completely laparoscopically, including free-hand laparoscopic suturing and in situ knot tying techniques. The mobilized specimens were removed in an endoscopy bag by way of the rectum or vagina. RESULTS: All operations were completed laparoscopically by two surgeons without conversion to open surgery. The median operating time was 485 minutes. One patient needed a blood transfusion of 2 U. All specimens had negative surgical margins. Two patients required reoperation. The median follow-up was 33 months. All patients were continent during the day, and 11 patients were continent at night. One patient developed unilateral hydronephrosis with loss of kidney function. No patient developed local recurrence, but 3 patients had systemic progression. Two of the three died of metastatic disease. CONCLUSIONS: This is the first series of laparoscopic radical cystectomy using an intracorporeal rectosigmoid pouch. Our functional data for continence and upper urinary tract obstruction were comparable with those of open rectosigmoid pouch studies. We were able to demonstrate an oncologic outcome similar to that achieved by the open surgical approach. PMID- 15533482 TI - Comparison of modified Taguchi and Bricker ureteral reimplantation techniques after radical cystectomy. AB - OBJECTIVES: To present our experience with the modified Taguchi "single-stitch" ureteral reimplantation technique in patients undergoing radical cystectomy with urinary diversion compared with a traditional Bricker reimplantation technique. Improved techniques are continually sought for ureteroenteric anastomoses during urinary diversion. The modified Taguchi "single-stitch" ureteral reimplantation is reportedly a time-efficient technique that preserves anastomotic integrity. METHODS: We retrospectively examined 75 consecutive patients with bladder cancer who underwent cystectomy and urinary diversion between October 1, 1999 and March 31, 2001. The ureteroenteric anastomosis was performed using a reinforced single stitch modified Taguchi technique in the first 36 patients and an interrupted two layer Bricker technique in the subsequent 39 patients during the creation of 47 orthotopic neobladders and 28 ileal conduit diversions. Ureteral stents were not routinely used. The demographic and perioperative clinical parameters were evaluated in each cohort, with particular attention to ureteral complications. RESULTS: Modified Taguchi and Bricker ureteral anastomoses were performed in 48% and 52% of patients, respectively. Patient age, sex, and body mass indexes were similar between groups. Apart from pathologic stage, univariate analysis did not demonstrate statistically significant differences between the groups in the demographic, intraoperative (estimated blood loss, diversion type, operative time) or postoperative (length of stay, rate of complications) parameters. Ureteral complications occurred in 8% of the Bricker group and 15% of the modified Taguchi group (P = 0.23). CONCLUSIONS: Either technique can be performed safely and in a timely fashion. However, the increased number of ureteral leaks in the modified Taguchi cohort combined with no advantage in procedure time, prompted our return to the Bricker technique exclusively. PMID- 15533483 TI - Comparisons of urodynamic characteristics between female patients with overactive bladder and overactive bladder plus stress urinary incontinence. AB - OBJECTIVES: To determine and compare the urodynamic characteristics in patients with overactive bladder (OAB) and patients with OAB plus stress urinary incontinence (OAB+SUI). METHODS: A total of 120 patients (60 each in OAB and OAB+SUI groups) who underwent urodynamic study between January and April 2003 were recruited. A detailed history, physical examination, and multichannel urodynamic data, including uroflowmetry, filling and voiding cystometry, stress urethral pressure profile, and 20-minute pad test were obtained for each patient. The urodynamic findings of each patient were analyzed and the results compared between the two groups. RESULTS: The median age of both groups was 51 years (interquartile range 43 to 64) with a parity of 3 (interquartile range 2 to 4); 54% (n = 65) were menopausal. The urodynamic parameters of bladder storage function showed premature filling sensation with decreased bladder capacity. Additionally, 60 (50%) had genuine stress incontinence, 3 (3%) had idiopathic detrusor overactivity, 11 (9%) mixed type incontinence, and 12 (11%) had voiding dysfunction. Among the urodynamic parameters between the two groups, first desire, strong desire, urgency, functional urethral length, maximal urethral pressure, maximal urethral closure pressure, pad test, and percentage of genuine stress incontinence in the OAB+SUI group were significantly lower statistically than those in the OAB group (P <0.03); the differences in age, parity, and percentage of menopausal status were also statistically significant (P <0.03). CONCLUSIONS: Our data showed that patients in the OAB+SUI group had more impaired urodynamic parameters than those in the OAB group. Age, parity, and menopausal status were contributory factors. PMID- 15533485 TI - Tranexamic acid in control of primary hemorrhage during transurethral prostatectomy. AB - OBJECTIVES: To determine whether short-term treatment of patients about to undergo transurethral resection of the prostate (TURP) with tranexamic acid (TXA) would be beneficial in reducing the associated blood loss. METHODS: A prospective and randomized trial was conducted with 136 men requiring TURP for obstructive urinary symptoms. The treatment group received 2 g TXA three times daily on the day of, and first day after, the operation. RESULTS: Short-term TXA treatment significantly reduced the operative blood loss associated with TURP (128 mL versus 250 mL, P = 0.018), and this difference was not a result of the amount of tissue resected between the two groups (16 g versus 16 g, P = 0.415). In addition, TXA treatment reduced the amount of blood loss per gram of resected tissue (8 mL/g versus 13 mL/g, P = 0.020). Furthermore, the volume of irrigating fluid required (15 L versus 18 L, P = 0.004) and operating time (36 minutes versus 48 minutes, P = 0.001) were also reduced. However, TXA treatment did not influence the number of patients requiring a blood transfusion. Six patients in the treatment group (7.2%) and five in the control group (6.8%) required a transfusion (P = 0.709). Moreover, TXA treatment did not affect the duration of catheterization (1 day versus 1 day, P = 0.342) or hospitalization (3 days versus 3 days, P = 0.218). CONCLUSIONS: Short-term TXA treatment is effective in reducing the operative blood loss associated with TURP. PMID- 15533484 TI - Urethral recurrence after cystoprostatectomy: implications for urinary diversion and monitoring. AB - OBJECTIVES: To review our cystoprostatectomy (CP) database to determine the urethral recurrence rate. Urethral recurrence after CP has been reported to occur in up to 10% of patients. Recent data have suggested a much lower incidence. This has important implications when considering the type of urinary diversion and postoperative monitoring. METHODS: We retrospectively analyzed our single surgeon, consecutive CP series and determined the urethral recurrence rate and prognostic factors for recurrence. Urethrectomy was performed at CP if the prostatic apical margin was positive for carcinoma. All patients were followed up quarterly for 2 years and then semiannually. Urethral wash cytology was obtained if the patient had an ileal conduit. Cytology and cystoscopy were performed if they had an orthotopic neobladder. RESULTS: A total of 226 men had undergone radical CP. The mean age for all patients was 69 years. Eight (3.5%) had undergone urethrectomy at CP. The mean follow-up was 42 months for the remaining 218 patients, of whom 108 had an orthotopic neobladder and 110 had supravesical diversion. Of the 218 patients, 8 (3.7%) developed urethral recurrence, 7 (6.4%) in the 110 who had undergone supravesical diversion and 1 in the 108 (0.9%) who had an orthotopic neobladder. Seven patients underwent urethrectomy for the recurrence and had no evidence of disease at last follow-up. One patient died of metastatic transitional cell carcinoma at 61 months. CONCLUSIONS: In our series, the risk of urethral recurrence after radical CP was low. The risk was substantially lower for patients who had an orthotopic neobladder. Our results show that urethrectomy at CP is rarely necessary because the proximal urethral margin is usually free of cancer. An orthotopic neobladder can therefore be safely considered in most patients. Delayed urethrectomy can be safely performed in those few patients with isolated urethral recurrence without compromising their survival. PMID- 15533486 TI - Longitudinal changes in lower urinary tract symptoms among a cohort of black American men: the Flint Men's Health Study. AB - OBJECTIVES: To examine the progression of lower urinary tract symptoms in a longitudinal population-based cohort of black men. Population-based studies of prostatism and longitudinal data regarding changes in lower urinary tract symptom severity have largely focused on white men, with little attention directed toward black men. METHODS: In 1996, a probability sample of 369 black men, aged 40 to 79 years, residing in Genesee County, Michigan, and without a prior history of prostate cancer/surgery participated in a prostate cancer screening protocol that included completing the American Urological Association Symptom Index (AUASI). Four years after baseline, 175 of the 369 men agreed to participate in the follow up protocol. Of the 175 men, 149 had not reported undergoing treatment for benign prostatic hyperplasia and had complete symptom data. These men were included in this study. Differences between baseline and follow-up AUASI scores were examined. RESULTS: The mean and standard deviation AUASI scores at baseline and follow-up were 7.1 (6.4) and 7.0 (6.8), respectively. Although overall no statistically significant change was found in the mean AUASI during the 4 years of follow-up (-0.11; SD 6.2; P = 0.7), the average change in the symptom score and the variability in the change increased with patient age at baseline from a mean of -0.42 (SD 5.0) among men in their 40s to 2.1 (SD 6.6) among men in their 70s. Of the 91 men (61.1%) who reported mild to no symptoms (AUASI score 7 or less) at baseline, 24 (26.4%) reported moderate to severe symptoms (AUASI score 8 or more) at follow-up. This progression of symptom severity was observed across all ages. CONCLUSIONS: In this population-based study of longitudinal changes in urinary symptoms in black men, we found a substantial percentage of men demonstrated a measurable progression in urinary symptom severity over time. Additional studies are needed to examine critically any racial differences in lower urinary tract symptom progression. PMID- 15533487 TI - Is questionnaire enough to assess number of nocturic episodes? Prospective comparative study between data from questionnaire and frequency-volume charts. AB - OBJECTIVES: To determine whether the voiding symptom questionnaire provides information regarding nocturia as accurate as that provided by frequency-volume (FV) charts, we evaluated patients with lower urinary tract symptoms. METHODS: We prospectively analyzed 164 patients (62 men and 102 women, mean age 53.6 years, range 20 to 83) referred for the evaluation of lower urinary tract symptoms. At the initial visit, all patients answered the International Prostate Symptom Score (IPSS) questionnaire to quantify nocturia and subsequently were requested to complete a 3-day FV chart. RESULTS: The differences in age between the men and women were not statistically significant. The total IPSS and IPSS bother scores were similar between the sexes. For all patients, the actual number of nightly voids in the FV charts and the IPSS nocturia score were modestly intercorrelated (r = 0.609, P <0.001). However, large individual differences were found in the nighttime diuresis rate (67.7%) between the IPSSs and FV charts. No statistically significant differences were found between men and women, but agreement was achieved in only 39.2% of patients younger than 60 years old and 22.4% of those 60 years old or older for the number of voids per night recorded by the FV charts and reported by the IPSS (P = 0.024). Overestimation of the number of nocturic episodes in the IPSS was more common than underestimation, especially in patients 60 years old or older (P = 0.014). CONCLUSIONS: The findings of the present study revealed poor agreement between the subjectively estimated nocturnal frequency and chart-determined nocturnal frequency. Our results suggest that FV charts should be included as an integral part of evaluating nocturia in both sexes. PMID- 15533488 TI - Adherence to a low-fat diet in men with prostate cancer. AB - OBJECTIVES: To evaluate, in a feasibility study, the adherence to a low-fat diet by men with prostate cancer. Evidence is growing that a low-fat diet affects the development and course of prostate cancer. To design preventive and therapeutic interventions, it is important to know whether men will adhere to these nutritional recommendations, particularly when motivated by the diagnosis of prostate cancer. METHODS: Men with elevated prostate-specific antigen levels, most of whom were recently treated for prostate cancer, were randomized to one of four dietary regimens for which they received nutritional counseling: a low-fat diet (15% fat or less) with supplements (vitamin E and selenium), a low-fat diet (15% fat or less) without the supplements, the supplements alone, and a control group. Adherence was evaluated by the change in weight, fat intake, free fatty acids, cholesterol, high-density and low-density lipoproteins, and triglycerides during a 12-month period. RESULTS: The mean age of the 48 participants was 66 years. For those counseled about a low-fat diet, the mean change in the percentage of energy (kilocalories) in the diet from fat was greater after 3 months (-8.6% versus +2.1%, P <0.001) and 12 months (-9.8% versus -1.6%, P = 0.001). Three months after starting the intervention, those randomized to low-fat dietary counseling had lost 2 kg, on average, compared with 0.8 kg lost by those who did not receive this counseling (P = 0.09). At 12 months, those receiving low fat counseling had lost 2.8 kg, on average, compared with 0.5 kg gained among the other groups (P = 0.02). CONCLUSIONS: With appropriate counseling, men with prostate cancer can adhere to a low-fat dietary intervention for a 12-month period. PMID- 15533489 TI - Androgen ablation adjuvant to postprostatectomy radiotherapy: complication adjusted number needed to treat analysis. AB - OBJECTIVES: To quantify the benefits and harm of androgen ablation (AA) adjuvant to radiotherapy in the postprostatectomy setting. AA is commonly used in the management of prostate cancer. METHODS: A literature review was performed to estimate the absolute biochemical control advantage for the use of AA concomitant with postprostatectomy external beam radiotherapy. Additionally, a model was developed, with supporting published data, to estimate the utility-adjusted survival detriment due to the side effects of AA, using the number needed to treat (NNT) technique. Using these data, the unadjusted NNTs and the utility adjusted NNTs for the addition of AA were computed. In all cases, the sign and magnitude of the NNTs obtained were used to gauge the effects of AA. RESULTS: The unadjusted NNT analysis demonstrated very low values (far less than 20), suggesting a strong benefit for the use of AA, in both adjuvant and salvage radiotherapy settings. Even after adjustment for hormone-induced functional loss, a significant advantage of AA was demonstrated. CONCLUSIONS: Using the complication-adjusted NNT method, AA appears to be advantageous in both adjuvant and salvage postprostatectomy radiotherapy settings. The results of the present investigation demonstrated the significant role of the NNT technique for uro oncologic management decisions when treatment complications need to be considered and balanced against the beneficial effects of the treatment. PMID- 15533490 TI - Postoperative adjuvant and salvage radiotherapy for prostate cancer: impact on freedom from biochemical relapse and survival. AB - OBJECTIVES: To determine the therapeutic outcomes in patients with high-risk prostate cancer treated with adjuvant or salvage radiotherapy (RT) after radical prostatectomy. METHODS: Between 1982 and 2000, 163 patients were treated with RT after radical prostatectomy. Adjuvant therapy was administered to 107 consecutive node-negative patients (T2-T4N0) referred to our institution less than 1 year after surgery for postoperative RT. Salvage treatment was delivered to 56 patients for a persistently elevated prostate-specific antigen level, biochemical relapse after surgery, or local recurrence. RESULTS: The median follow-up was 70 months (range 2 to 167) from the initiation of RT. Patients treated with adjuvant RT were less likely than those treated with salvage RT to experience biochemical relapse. At 5 and 10 years, the rate of freedom from biochemical relapse was 80% and 66% in the adjuvant cohort compared with 39% and 22% for patients treated with salvage intent, respectively (P <0.0001). This did not translate into a statistically significant improvement in absolute survival (72% versus 70%) or cause-specific survival (93% versus 86%) at 10 years. On multivariate analysis, neoadjuvant hormonal therapy (P = 0.0187), presence of seminal vesicle involvement (P = 0.0002), and referral indication for postoperative RT (salvage versus adjuvant RT; P <0.001) were predictors of biochemical relapse. CONCLUSIONS: In this single-institution experience, patients at high risk of disease recurrence after radical prostatectomy realized a greater biochemical relapse-free survival benefit when treated with adjuvant RT than with salvage RT. Neoadjuvant hormonal therapy and seminal vesicle involvement predicted for inferior treatment outcome. PMID- 15533491 TI - Effect of methylprednisolone on return of sexual function after nerve-sparing radical retropubic prostatectomy. AB - OBJECTIVES: To determine whether postoperative methylprednisolone improves the recovery of sexual function after nerve-sparing radical retropubic prostatectomy. METHODS: We randomized men undergoing bilateral nerve-sparing radical retropubic prostatectomy by a single surgeon to receive 6 days of placebo or methylprednisolone beginning on postoperative day 1. At 3, 6, and 12 months postoperatively, we assessed potency with the abbreviated International Index of Erectile Function questionnaire and urinary continence with participant-reported pad use. We used the chi-square test, Fisher's exact test, and the two-sample t test with equal variances for comparisons between study groups. RESULTS: No operative complications occurred and 70 (100%) of 70 participants experienced normal wound healing. The odds of being potent for participants who received methylprednisolone (n = 34) compared with those who received placebo (n = 36) did not significantly differ at 3 (odds ratio 0.29, 95% confidence interval 0.08 to 1.05), 6 (odds ratio 0.63, 95% confidence interval 0.17 to 2.4), or 12 (odds ratio 1.18, 95% confidence interval 0.29 to 4.8) months. The mean International Index of Erectile Function scores did not significantly differ at 3 (P = 0.08), 6 (P = 0.50), or 12 (P = 0.71) months. At 12 months, 74% of the methylprednisolone and 71% of the placebo participants were potent (P = 0.8). The proportions of participants who were continent did not differ significantly at 3 (P = 0.89), 6 (P = 0.25), or 12 (P = 0.49) months. At 12 months, 96% of the methylprednisolone and 100% of the placebo participants were continent. CONCLUSIONS: At doses sufficient to produce a systemic anti-inflammatory effect, postoperative methylprednisolone was not associated with improved potency at up to 12 months after bilateral nerve-sparing radical retropubic prostatectomy in men 40 to 60 years old. PMID- 15533492 TI - Sexual behavior and sexual dysfunctions after age 40: the global study of sexual attitudes and behaviors. AB - OBJECTIVES: To assess the importance of sex and the prevalence of sexual dysfunction among middle-aged and older adults throughout the world. Increasing life expectancy has been accompanied by improvements in the health of the middle aged and elderly, but little is known about how this has affected their sexual experience. METHODS: Data were collected in 29 countries from 27,500 men and women aged 40 to 80 years using a standardized questionnaire (self-completed or by interview). Sexual dysfunction was defined as frequent and persistent problems. They included early ejaculation and erectile difficulties in men, lubrication difficulties and pain during intercourse in women, and a lack of sexual interest, an inability to achieve orgasm, and a feeling of unpleasurable sex in both. RESULTS: More than 80% of the men and 65% of the women had had sexual intercourse during the past year. Of these subjects, the most common dysfunctions were early ejaculation (14%) and erectile difficulties (10%) among the men and a lack of sexual interest (21%), inability to reach orgasm (16%), and lubrication difficulties (16%) among the women. Overall, 28% of the men and 39% of the women said that they were affected by at least one sexual dysfunction. CONCLUSIONS: The results of our study indicate that sexual desire and activity are widespread among middle-aged and elderly men and women worldwide and persist into old age. The prevalence of sexual dysfunctions was quite high and tended to increase with age, especially in men. Although major between-country differences were noted, this global study revealed some clear and consistent patterns. PMID- 15533493 TI - Simultaneous administration of vardenafil and tamsulosin does not induce clinically significant hypotension in patients with benign prostatic hyperplasia. AB - OBJECTIVES: To assess the pharmacodynamic effects of coadministered vardenafil and tamsulosin in patients with benign prostatic hyperplasia (BPH) undergoing stable tamsulosin therapy. METHODS: In this Phase 1, placebo-controlled, two stage, two-way, crossover study, 22 patients undergoing stable (longer than 4 weeks) tamsulosin therapy for BPH (18 using 0.4 mg and 4 using 0.8 mg tamsulosin daily) received vardenafil 10 mg (or placebo), followed by vardenafil 20 mg (or placebo), simultaneously with tamsulosin. The mean maximal change from baseline with vardenafil use versus placebo was evaluated for supine and standing blood pressure and heart rate for up to 6 hours after dosing. RESULTS: In patients receiving vardenafil 10 mg, the mean maximal change from baseline versus placebo in supine systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate was -4.5 mm Hg (95% confidence interval [CI] -8.2 to -0.8), -2.3 mm Hg (95% CI -4.9 to 0.4), and 3.7 beats per minute (95% CI 1.1 to 6.3), respectively. In patients receiving vardenafil 20 mg, the mean maximal change from baseline versus placebo in supine SBP, DBP, and heart rate was -4.0 mm Hg (95% CI -6.3 to 1.8), -2.9 mm Hg (95% CI -5.6 to -0.2), and 0.8 beats per minute (95% CI -1.2 to 2.9), respectively. These hemodynamic changes were similar to those obtained in the standing position. Two placebo patients and 1 vardenafil 10-mg patient had a drop of 20 mm Hg or more in standing DBP; 1 vardenafil 10-mg patient had a standing SBP drop of 30 mm Hg or more. No patient exhibited symptomatic hypotension (SBP less than 85 mm Hg with dizziness). Three patients receiving vardenafil 20 mg/tamsulosin 0.4 mg reported dizziness, but never had an SBP of less than 95 mm Hg. No serious adverse events were reported. CONCLUSIONS: In this study, no evidence was found that coadministered vardenafil and tamsulosin induced clinically significant hypotension in patients with BPH. PMID- 15533495 TI - Subinguinal microvaricocelectomy versus retroperitoneal varicocelectomy: comparative study of complications and surgical outcome. AB - OBJECTIVES: To compare and assess objectively the postoperative outcome parameters of both microsurgical subinguinal artery sparing and retroperitoneal high ligation techniques for varicocele in infertile men. METHODS: We performed a prospective study that included 413 consecutive patients who presented with varicocele. Of the 413 patients, 304 underwent subinguinal varicocelectomy (group 1) and 109 patients underwent high ligation varicocelectomy (group 2). Subinguinal varicocelectomies were performed under local anesthesia, with intravenous propofol sedation administered as needed. The operating microscope (x10 to x16) was used to spare the arteries and lymphatics. High ligation was performed through a retroperitoneal approach. RESULTS: All patients went home on the evening after surgery with minimal discomfort. No intraoperative complications occurred. The internal spermatic artery was identified in all microvaricocelectomy cases. No testicular atrophy occurred in either group. Five (1.6%) and seven (6.4%) hydroceles were identified at the 3-month postoperative visit in groups 1 and 2, respectively. CONCLUSIONS: Microvaricocelectomy is safe and effective and associated with a rapid recovery and minimal morbidity. However, it requires microsurgical training. PMID- 15533496 TI - Relationship of interleukin-6 with semen characteristics and oxidative stress in patients with varicocele. AB - OBJECTIVES: To examine levels of interleukin-6 (IL-6) in fertile semen donors and patients with varicocele and examine its association with semen characteristics and levels of reactive oxygen species (ROS). METHODS: We conducted a prospective study consisting of 15 fertile donors (controls) and 35 infertile patients with varicocele. Semen analysis was performed according to the World Health Organization guidelines. IL-6 levels were measured using the enzyme-linked immunosorbent assay. ROS (x10(4) counted photons per minute per 20 x 10(6) sperm) and total antioxidant capacity (molar trolox equivalents) were measured using a chemiluminescence assay. RESULTS: The sperm concentration and motility were significantly greater in the donors compared with the infertile patients with varicocele (P <0.0001 and P = 0.01, respectively). The IL-6 (log10 [IL-6 +1]) and ROS (log10 [ROS +1]) levels were significantly greater in infertile patients with varicocele than in the donors (IL-6: 2.1 [1.7, 2.4] versus 0.7 [0, 1.9], P = 0.003; ROS: 1.8 [1.2, 2.6] versus 1.0 [0.7, 1.6], P = 0.04). The total antioxidant capacity levels were significantly lower in the varicocele patients (1166.7 +/- 366.2) than in the donors (1556.4 +/- 468.1; P = 0.003). The IL-6 levels correlated significantly with the ROS levels in the infertile patients with varicocele (r = -0.39; P = 0.01). CONCLUSIONS: Infertile patients with varicocele exhibited elevated levels of IL-6 and ROS and decreased levels of total antioxidant capacity. Pro-inflammatory cytokine IL-6 and oxidative stress may play a role in the pathophysiology of infertility in these patients. PMID- 15533497 TI - Congenital midureteral stricture in children diagnosed with antenatal hydronephrosis. AB - OBJECTIVES: Antenatal hydronephrosis is the most common genitourinary pathologic finding during gestational ultrasonography. Congenital midureteral strictures are an unusual cause of prenatal hydronephrosis, with 13 total previously reported and only 1 diagnosed as a result of antenatal ultrasonography. METHODS: We present 4 cases of midureteral strictures as the cause of antenatal hydronephrosis and review the published data on congenital midureteral obstruction. RESULTS: All midureteral strictures in our series and in the published reports were treated surgically after varying periods of observation. Retrograde pyelography was diagnostic in all cases. CONCLUSIONS: This atypical cause of antenatal hydronephrosis may provide additional insight into the controversy regarding preoperative retrograde pyelography before surgical intervention for ureteropelvic junction obstruction. Given the typical misdiagnosis of ureteropelvic junction obstruction in the setting of midureteral stricture disease, we believe that the threshold for retrograde pyelography should be very low. PMID- 15533498 TI - Impact of urodynamics in treatment of primary nocturnal enuresis persisting into adulthood. AB - OBJECTIVES: To investigate the urodynamic profiles of adults with primary nocturnal enuresis (PNE) and the association of the urodynamic profile findings with the efficacy of desmopressin and/or tolterodine pharmacotherapy. At least 2% of adults are enuretic during the night. The diagnostic and treatment approach for PNE is empirically the same in children and adults. METHODS: A total of 20 nocturnal enuretic patients (12 women and 8 men) with a mean age of 27.1 years (range 20 to 42) were studied. They had wet their bed at least twice per week for the past 6 months. Urodynamic studies, including filling and voiding cystometry, pressure-flow study, and pelvic floor electromyography with superficial electrodes, were performed on all patients. Two of them had daytime symptoms, and two had prior failed desmopressin therapy. All patients began taking oral desmopressin 0.4 mg for 1 month. Their continence was assessed and tolterodine 4 mg was added for those in whom desmopressin alone failed. The patients responsive to desmopressin alone or desmopressin plus tolterodine were weaned from medication at 6 and 12 months to reassess continence. The mean follow-up period was 11.6 +/- 3.3 months (range 4 to 14). RESULTS: Urodynamic studies of 20 PNE adult patients revealed detrusor instability in 10 (50%), hypocompliance in 8 (40%), nonneurogenic detrusor-sphincter dyssynergy in 1 (5%), and no abnormality in 10 (50%). Of the 20 patients, 19 (95%) had no voiding bladder problems. Of the 10 patients responsive to desmopressin alone, 6 (60%) had a normal urodynamic profile; the remaining 4 (40%) had detrusor instability and/or hypocompliance. Of the 5 patients who received desmopressin and tolterodine, 3 achieved continence. The overall continence rate was 86% (13 of 15), and 12 (92%) of the 15 patients required maintenance therapy. In 2 patients (13.3%), desmopressin and tolterodine therapy failed. The efficacy of desmopressin alone and of desmopressin plus tolterodine were not related to the urodynamic profile findings (P >0.05). The urodynamic profile was also not related to the relapse rate after any form of pharmacotherapy (P >0.05). CONCLUSIONS: PNE persisting into adulthood may be associated with abnormal urodynamic findings. Patients may benefit from urodynamic studies, because if the findings are abnormal, they might have the best chance of successful treatment. PMID- 15533500 TI - Use of Amplatz sheath in male urethra during cystolitholapaxy of large bladder calculi. AB - Large-burden bladder stones often require percutaneous cystolithotomy or an open procedure. These approaches may not be feasible in certain patients who are at increased risk of operative complications. We present a novel technique for transurethral cystolitholapaxy using Amplatz sheath in the male urethra. PMID- 15533501 TI - An unusual varicocele due to spontaneous arteriovenous fistula. PMID- 15533502 TI - Nephrogenic adenoma of bladder after ibuprofen abuse. AB - Nephrogenic adenoma is an infrequent benign lesion of the urinary system that occurs in patients with a history of genitourinary surgery, stone disease, trauma, chronic urinary tract infection, or renal transplantation. We report the first case of nephrogenic adenoma of the bladder in a 53-year-old man with a 5 year history of ibuprofen abuse for chronic arthritis. We stress the importance of investigating the analgesic abuser for nephrogenic adenoma if microhematuria and/or irritative lower urinary tract symptoms are present. PMID- 15533503 TI - Laparoscopic injury and repair of obturator nerve during radical prostatectomy. AB - A 61-year-old man with bilateral Gleason score 7 (3+4) clinical Stage T1c prostate cancer was treated with laparoscopic bilateral pelvic lymphadenectomy and radical prostatectomy. The left obturator nerve was inadvertently transected during left obturator lymph node dissection and repaired by laparoscopic reapproximation. PMID- 15533504 TI - Treatment of voiding dysfunction after transobturator tape procedure. AB - Voiding dysfunction after mid-urethral sling procedures is uncommon but not completely avoidable. We report on a method to adjust the transobturator mid urethral sling under local anesthesia in the early postoperative period for postoperative voiding difficulty. At postoperative day 17 and 18, 2 women, who had undergone the transobturator tape procedure for stress incontinence, underwent successful loosening of the mesh through the previous vaginal incision, without the need to transect or remove the tape. Both patients had immediate resolution of their symptoms while maintaining urinary continence. PMID- 15533505 TI - Pelvic hematoma as a cause of bladder perforation and gross hematuria. AB - We present a case of an ulcerated perforation of the bladder caused by a large pelvic hematoma without any direct injury to the bladder in a 71-year-old woman. She developed the hematoma after emergency percutaneous angioplasty and placement of an intra-aortic balloon pump that was complicated by retroperitoneal bleeding from an injured femoral artery. She presented several days later with gross hematuria and right hydroureteronephrosis. Cystoscopy and cystography revealed an extraperitoneal perforation at the right bladder neck. Although conservative management was attempted, the patient eventually required open repair. The perforation had transformed to an intraperitoneal perforation by making a communicating tract through the absorbed hematoma. After debridement and excision of the tract, the bladder perforation was repaired in two layers. PMID- 15533506 TI - Urethral stricture after pancreas-kidney transplantation due to polypoid urethritis. AB - Urologic complications are common after pancreas-kidney transplantation using bladder drainage. We report a case of urethral stricture caused by polypoid urethritis occurring 4 years after simultaneous pancreas-kidney transplantation. Endoscopic exploration revealed irregular, ragged-appearing urethral mucosa. The diagnosis of polypoid urethritis was confirmed histopathologically. PMID- 15533507 TI - Modified pudendal thigh flap for perineoscrotal reconstruction: a case of Leriche syndrome with rapidly progressing Fournier's gangrene. AB - We present the first report of Leriche syndrome associated with Fournier's gangrene. We used a modified pudendal thigh flap in the treatment of an extensive perineoscrotal soft-tissue defect successfully. We propose this new robust flap as an addition to the existing reconstructive armamentarium and draw attention to the coexistence of Leriche syndrome and Fournier's gangrene. PMID- 15533508 TI - Bilateral ureteral compression by multiple abdominal desmoid tumors in Gardner syndrome: case report and literature review. AB - A 28-year-old man with Gardner syndrome was admitted to our Department because of multiple abdominal masses. Abdominal computed tomography revealed severe hydronephrosis of both kidneys due to ureteral compression against the pelvic bones exerted by multiple solid abdominal masses. The patient developed oligoanuria. Right percutaneous nephrostomy tube placement was followed by restoration of diuresis and progressive recovery of renal function of the left side. The patient then underwent median laparotomy and removal of five large desmoid tumors originating from the abdominal wall. Bilateral ureteral obstruction due to abdominal desmoid tumors can be a rare urologic complication of Gardner syndrome. PMID- 15533509 TI - Glomus tumor of the glans penis. AB - Glomus tumors of the penis are extremely rare. We report a patient with a solitary glomus tumor involving the penis. A 19-year-old man presented with a complaint of a recurrent painful penile mass. Resection of the lesion was performed. The pathologic diagnosis was glomus tumor of the glans penis. This case emphasizes the need for complete extirpation of the glomus tumor to avoid additional surgery. PMID- 15533510 TI - Testicular recurrence from "primary" retroperitoneal germ cell tumor. AB - Extragonadal germ cell tumors are a rare clinical entity. We present the case of an extragonadal germ cell tumor followed by testicular recurrence more than 4 years later. This presents an interesting dilemma as to whether this was recurrence of the original tumor or a new, primary tumor. The patient's outcome and surveillance protocol for these tumors are presented. PMID- 15533511 TI - Synchronous bilateral malignant mesothelioma of tunica vaginalis testis: early diagnosis. AB - We report a case of bilateral synchronous malignant mesothelioma of the testis. A 21-year-old man presented with an acute scrotum after 3 months of recurrent pain on the right side. Ultrasonography demonstrated an irregular thickening of the tunica vaginalis testis, scrotolyths, and an increased blood flow to the right epididymis testis. Bilateral exploration revealed torsion of the hydatid and a bilateral malignant mesothelioma of the tunica vaginalis. One year later, elective bilateral exploration showed one minute area on the parietal tunica vaginalis of malignant mesothelioma. After 2 years of follow-up, the patient had no signs of distant disease. PMID- 15533512 TI - Management of endosalpingiosis of urinary bladder. AB - We report on a 48-year-old woman who presented with abnormal uterine bleeding secondary to uterine fibroids and who was found to have incidental endosalpingiosis of the urinary bladder. Follow-up is necessary for bladder endosalpingiosis because of its tendency to recur. Follow-up should include cystoscopy and renal, and possibly pelvic, ultrasonography. PMID- 15533513 TI - Mutation analysis of INSL3 and GREAT/LGR8 genes in familial cryptorchidism. AB - OBJECTIVES: Male mice deficient in insulin-like 3 hormone (Insl3) or its receptor, Great/Lgr8, exhibit cryptorchidism. Recently, sequence analysis of the human INSL3 and GREAT genes identified several allelic variants. These include polymorphisms without apparent functional consequence and a few alleles encoding products with compromised function. However, loss-of-function alleles appear to be rare in human cryptorchidism. Most patients studied to date are presumed to have had sporadic cryptorchidism. We postulated that any genotypic variants predisposing to cryptorchidism would be more prevalent among patients with familial cryptorchidism. METHODS: We isolated genomic DNA from 13 individuals with personal and family histories of cryptorchidism and used polymerase chain reaction to amplify all exons of both INSL3 and GREAT, as well as INSL3 proximal promoter sequence, including a putative SF-1 transcription factor binding site. We directly sequenced all 20 amplicons and compared them with the wild-type alleles. RESULTS: We detected two silent substitutions and one missense (A60T) substitution in exon 1 of INSL3 and two silent substitutions in exon 12 and one missense (I604V) substitution in exon 17 of GREAT, all previously described. We found that in vitro the I604V GREAT variant receptor responds to INSL3 stimulation similarly to the wild-type receptor. CONCLUSIONS: We found polymorphic alleles of INSL3 and GREAT, but no deleterious mutations among individuals with familial cryptorchidism. Thus, mutations in these two genes are responsible only for a small proportion of familial cryptorchidism. PMID- 15533514 TI - Fate of implanted syngenic muscle precursor cells in striated urethral sphincter of female rats: perspectives for treatment of urinary incontinence. AB - OBJECTIVES: To analyze the outcome of syngenic skeletal muscle precursor cells (MPCs) after implantation in the striated urethral sphincter of the female rat. METHODS: MPCs were isolated from the striated muscles of the lower limbs and infected with a retrovirus carrying the gene for green fluorescent protein. Approximatively 10(5) cells were injected longitudinally in the striated urethral sphincter of 24 animals using a 10-muL Hamilton syringe. The whole urethra was excised at 0, 1, 7, 10, 14, 30, and 90 days after implantation for histologic study and fluorescence analysis of the transections. RESULTS: At days 0 and 1, some small, round, fluorescent MPCs were observed at the injection site. At day 7, significant MPC persistence was noted, with infiltration of inflammatory cells in the whole urethral wall (striated muscle layer, smooth muscle layer, and connective tissue). At day 10, some fusiform cells appeared in the striated muscle layer, suggesting the incorporation of MPCs into the striated myofibers. Inflammatory cells were no longer visible. At day 14, the fusiform cells tended to be larger. The small, round cells were no longer seen. At days 30 and 90, all myofibers of the striated muscle layer were strongly fluorescent, and no fluorescence was detectable in the smooth muscle layer. CONCLUSIONS: Implantation of skeletal MPCs in the urethral sphincter resulted in selective incorporation into striated myofibers. Muscle-derived cell autografting could represent a new approach for the treatment of urinary incontinence in humans. PMID- 15533515 TI - Prevention by 17beta-estradiol and progesterone of calcitonin gene-related peptide-induced elevation of skin temperature in castrated male rats. AB - OBJECTIVES: To clarify the relationship between calcitonin gene-related peptide (CGRP) and ovarian hormones (17beta-estradiol and progesterone) in hot flashes in men who undergo androgen deprivation therapy for prostate cancer, we studied the effects of ovarian hormones on CGRP-induced elevation of skin temperature in castrated male rats. The results were compared with those from rats treated with testosterone replacement. METHODS: Adult male rats were castrated by either a single injection of gonadotropin-releasing hormone analogue (Leuplin, 1.0 mg/kg, subcutaneously) or bilateral orchiectomy. The castrated animals were subcutaneously injected daily for 14 days with ovarian hormones, testosterone, or olive oil as the vehicle. On the day after the final administration of the drug, the changes in skin temperature induced by exogenous CGRP (10 mug/kg intravenously), serum testosterone concentration, and prostate weight were measured. RESULTS: The CGRP-induced elevation of skin temperature was significantly greater in the castrated rats than in the sham-treated rats. This potentiation was significantly inhibited by treatment with ovarian hormones, as well as by testosterone replacement. The testosterone replacement restored decreases in both the serum testosterone level and the prostate weight due to castration; the treatment with ovarian hormones did not affect them. CONCLUSIONS: 17beta-Estradiol and progesterone, which do not confer testosterone activity on serum, may be useful for the treatment of hot flashes in patients for whom testosterone replacement therapy is contraindicated, such as those with prostate carcinoma. In addition, we suggest that CGRP is closely involved in the amelioration of hot flashes by ovarian hormones in men who undergo androgen deprivation therapy. PMID- 15533516 TI - Effects of castration on female rabbit bladder physiology and morphology. AB - OBJECTIVES: To determine the effect of ovariectomy on bladder blood flow and oxygen tension. Women are subject to bladder dysfunctions that occur less frequently in men. These include interstitial cystitis (syndrome of urgency, frequency, and pain on distension), incontinence, and bladder infections. It is believed that alterations in female sex hormones play a major role in mediating these abnormalities. We believe that alterations in estrogen can have marked effects on the blood flow to the bladder. METHODS: We divided 20 female rabbits into two equal groups: ovariectomized and sham operated. Six weeks later, the bladder of each rabbit was evaluated for bladder capacity, compliance, permeability, blood flow, tissue hypoxia, morphology, and smooth muscle contraction. RESULTS: Ovariectomy resulted in decreased bladder compliance; decreased blood flow to the bladder mucosa and uterus, a smaller decrease in blood flow to the bladder smooth muscle, and mucosal hypoxia; and statistically significant thinning of the bladder mucosa and increased mucosal permeability. CONCLUSIONS: Ovariectomy resulted in decreased blood flow and hypoxia to the bladder mucosa and a mildly decreased blood flow to the smooth muscle. The decreased blood flow and hypoxia may be related to the age-related uropathologic findings in postmenopausal women. PMID- 15533517 TI - Association of bladder tumors and GA genotype of -308 nucleotide in tumor necrosis factor-alpha promoter with greater tumor necrosis factor-alpha expression. AB - OBJECTIVES: To investigate whether the genetic polymorphism of the -308 nucleotide in the tumor necrosis factor-alpha (TNF-alpha) promoter is associated with bladder cancer. METHODS: DNA samples from blood and tumor were analyzed by polymerase chain reaction-based restriction fragment length polymorphism to characterize the genetic polymorphism of the -308 nucleotide in the TNF-alpha promoter. TNF-alpha mRNA expression levels were assessed by quantitative competitive polymerase chain reaction, and the serum concentrations of TNF-alpha were measured using enzyme-linked immunosorbent assay. RESULTS: Patients with bladder tumor and control subjects did not differ in their genetic polymorphism of the -308 nucleotide (P = 0.259). However, the relation of the tumor grade with the GA phenotype was statistically significant (P = 0.04). TNF-alpha mRNA concentrations were also significantly greater in the GA genotype than in the GG genotype (P = 0.022). The TNF-alpha serum levels of the GA genotype were significantly greater than those of the GG genotype for both patients and controls. However, patients with bladder tumor had significantly greater TNF alpha serum levels than did the controls for both the GG and the GA genotypes (GG type, P = 0.001; GA type, P = 0.009). CONCLUSIONS: The genotype of the -308 nucleotide in the TNF-alpha promoter had a statistically significant effect on TNF-alpha production and was related to the bladder tumor grade. The GA polymorphism might be associated with a statistically significant increase in gene transcription. That the TNF-alpha serum levels were greater in the patients with bladder tumor compared with controls suggests that high TNF-alpha production is associated with bladder tumor development and that bladder tumors may secrete TNF-alpha. PMID- 15533518 TI - Prognostic significance of oxidative DNA damage evaluated by 8-hydroxy-2' deoxyguanosine in patients undergoing radical nephrectomy for renal cell carcinoma. AB - OBJECTIVES: To determine whether the degree of oxidative DNA damage in renal cell carcinoma (RCC) can be used as a useful prognostic predictor for patients who undergo radical nephrectomy. METHODS: We measured 8-hydroxy-2'-deoxyguanosine (8 OHdG), one of the most commonly used markers for evaluating oxidative stress, in DNA isolated from 72 RCC specimens, as well as adjacent normal kidney tissue using a quantitative sandwich enzyme-linked immunosorbent assay. RESULTS: The mean value of 8-OHdG in RCC was significantly greater than that in the adjacent normal tissue. The level of 8-OHdG in RCC compared with that in normal tissue (8 OHdG ratio) was significantly associated with other prognostic parameters, including mode of detection, maximal tumor size, distant metastasis, pathologic stage, tumor grade, and microscopic venous invasion. Furthermore, cancer-specific survival in patients with an elevated 8-OHdG ratio was significantly lower than that in patients with a normal 8-OHdG ratio; however, multivariate analysis using a Cox proportional hazards model showed that maximal tumor size and distant metastasis could be used as independent predictors of cancer-related death. CONCLUSIONS: These findings suggest that despite the lack of independent significance, the 8-OHdG ratio could be a useful prognostic indicator for patients with RCC; therefore, careful follow-up should be considered in those with an elevated 8-OHdG ratio. PMID- 15533519 TI - Oxidative stress plays a role in diabetes-induced bladder dysfunction in a rat model. AB - OBJECTIVES: To evaluate the oxidative status of the bladder 8 weeks after diabetes induction. Oxidative stress has recently been implicated in the pathogenesis of diabetes complications, but its role in diabetic cystopathy has not been studied. METHODS: Sprague-Dawley rats were divided into three groups: control (n = 11), diuretic control (5% sucrose drink; n = 6), and streptozotocin induced diabetic group (n = 14). Eight weeks later, the bladders were dissected. We measured the antioxidant scavenging enzymes (catalase and superoxide dismutase)-like activity and the levels of the thiobarbituric acid reactive substances, as a marker of lipid peroxidation. We also examined the levels of inducible nitric oxide synthase and apoptosis in the bladders. RESULTS: We found a statistically significant reduction in the catalase-like activity in the bladders from the diabetic group compared with the other groups (P = 0.017, diabetic versus control); the difference in the superoxide dismutase-like activity was not statistically significant among the groups. The thiobarbituric acid reactive substances levels were significantly greater in the diabetic compared with other groups (131.9 +/- 47.5, 46.7 +/- 17.9, and 60.9 +/- 25.4 nmol/mg protein in the diabetic, control, and diuretic group, respectively, P = 0.006, diabetic versus control). Immunohistochemical and apoptosis studies showed a statistically significant increased number of inducible nitric oxide synthase positive cells and apoptotic cells in the diabetic bladder smooth muscle cells (P <0.001). CONCLUSIONS: Our findings showed that oxidative stress occurred in the bladders of the STZ-diabetic rats and was not mediated by diuresis. The oxidative damage of the smooth muscle cells may be a contributory factor in diabetic cystopathy. PMID- 15533520 TI - Physical activity promotion through the mass media: inception, production, transmission and consumption. AB - BACKGROUND: Evaluations of physical activity and health media campaigns have been limited and ignore the complex process of communication and the socially constructed nature of news messages. METHODS: A systematic search strategy was conducted of the literature which was then assessed from two perspectives. First, studies since 1998 were reviewed for their success in impacting message recall and behavior change. Second, employing a critical media studies perspective the papers were assessed for the presence of a more sophisticated understanding of the media processes of inception, transmission and reception. RESULTS: Overall, recent studies support mass media interventions in influencing short-term physical activity message recall and to a lesser extent associated changes in physical activity knowledge. However, the majority of the papers were found to follow a social marketing or media advocacy theory of media promotion with little in-depth consideration of the comprehensive media processes involved in creating media messages and meaning. CONCLUSIONS: Simplistic understandings of media transmission dominate in assessing physical activity and health media campaigns. Fuller understandings of the success of media campaigns, the recall of media messages or associated behaviour change can only truly be understood through the application of a more sophisticated form of media analysis. PMID- 15533521 TI - Financial impact of a comprehensive multisite workplace health promotion program. AB - INTRODUCTION: The purpose of this study is to determine if the Washoe County School District Wellness Program impacted employee health care costs and rates of absenteeism over a 2-year period. METHODS: Outcome variables included health care costs and absenteeism during 2001-2002. Data were collected on 6246 employees over a 6-year period from 1997-2002. Baseline health claims costs and absenteeism from 1997-2000, age, gender, job classification, and years worked at the school district were treated as covariates. Logistic regression was used to compare 2 year costs and absenteeism rates between nonparticipants and employees who participated for 1 and 2 years. RESULTS: No significant differences in health care costs were found between those who participated in any of the wellness programs and those who did not participate. There was a significant negative association between participation and absenteeism; program participants averaged three fewer missed workdays than those who did not participate in any wellness programs. The decrease in absenteeism translated into a cost savings of 15.60 US dollars for every dollar spent on the program. CONCLUSIONS: After controlling for several confounding variables, wellness program participation was associated with large reductions in employee absenteeism. PMID- 15533522 TI - Effect of smoking on hearing loss: quality assessment and meta-analysis. AB - BACKGROUND: There is an accumulating body of research showing that smoking causes hearing loss; however, the results of these studies have been inconsistent. METHODS: Original English articles were retrieved by MEDLINE search using key words "smoking" and "hearing" (1966-2003). Of 166 relevant studies, those that investigated the risk for hearing loss in smokers identified by pure-tone average were selected for review. Studies with an occupational noise-exposed population were excluded. Methodological quality was assessed by a standardized checklist, and then a meta-analysis was performed on studies with discrete numbers of hearing loss among smokers and nonsmokers. RESULTS: A total of 15 (10 cross sectional, 4 cohort, and one case-control) observational studies were identified; the quality scores of the 9 studies with positive associations between smoking and hearing loss were comparable to those of the remaining 6 studies with insignificant associations. Concerning eight analyzable studies, risk ratios (95% confidence intervals) for hearing loss in smokers were 1.33 (1.24, 1.44) for cross-sectional studies, 1.97 (1.44, 2.70) for cohort studies, and 2.89 (2.26, 3.70) for case-control studies, respectively. CONCLUSIONS: The evidence was suggestive of a positive association between smoking and hearing loss. It is possible that smoking cessation may be a useful strategy for maintaining hearing acuity. PMID- 15533523 TI - Physical activity, dietary intake and metabolic risk factors in non-diabetic daughters of patients with type II diabetes. AB - BACKGROUND: It has been reported that the offspring of patients with type II diabetes have an adverse metabolic risk profile. This study aimed to investigate the impact of habitual physical activity and diet on metabolic risk factors in the daughters of patients with type II diabetes and control subjects. METHODS: Thirty-nine offspring and 39 age- and sex-matched controls completed physical activity and food intake diaries, during the week preceding a fasting blood sample. RESULTS: The offspring had higher body mass index, percentage body fat, and waist circumference than the control subjects (all P < 0.01). Fasting glucose and insulin, and insulin sensitivity estimated by the homeostasis model assessment (HOMA(IR)) method, were also higher in the offspring group (all P < 0.01). Daily energy expenditure was lower (P < 0.0001) in the offspring than control group. Dietary profile was not different between the groups. Daily energy expenditure was significantly correlated with waist circumference, fasting insulin, and HOMA(IR) (all P < 0.05) in offspring but not controls. CONCLUSIONS: Offspring had a less favourable physical and metabolic profile and were less physically active than control subjects. In offspring, central adiposity and metabolic risk factors were influenced by habitual physical activity to a greater degree than in control subjects. PMID- 15533524 TI - Physician practices and attitudes regarding adult immunizations. AB - OBJECTIVES: To assess immunization practices and attitudes of U.S. primary care physicians regarding adult influenza and pneumococcal immunizations. METHODS: Mailed survey of primary care internists and family physicians across the United States; four follow-up contacts by mail and telephone. Bivariate and multivariate analyses assessed immunization practices and attitudes and differences by physician characteristics. RESULTS: Three hundred and sixteen of 668 eligible physicians responded (50 refused, response rate of 266 = 40%); 220 provided adult vaccinations. More than 64% indicated they routinely vaccinated patients >/=65 years and those <65 years with chronic disease indications with both influenza and pneumococcal vaccine. Reported barriers for influenza vaccination included vaccine safety concerns by patients (58%), urgent concerns dominating visits (43%), and inadequate reimbursement (26%). Reported barriers for pneumococcal vaccination included urgent concerns during office visits (44%), no patient immunization history (36%), patient concerns about vaccine safety (31%), and inadequate reimbursement (25%). Many physicians indicated willingness to try tracking systems (72%), chart reminders (55%), patient reminders (53%), standing orders (36%), external lists of unimmunized patients for pneumococcal vaccination (74%), external patient reminders (70%), and office training of physicians (36%) or staff (46%). CONCLUSIONS: While most physicians favored adult vaccinations, practical barriers to vaccination exist. Most physicians would adopt evidence based strategies to improve immunization delivery. PMID- 15533525 TI - Appropriate antibiotic use: variation in knowledge and awareness by Hispanic ethnicity and language. AB - BACKGROUND: Recent campaigns are informing the public that antibiotics are inappropriate for viral respiratory infections. As little is known about their effect on populations challenged by less access to care, lower education, low income, low English proficiency, or non-mainstream cultural backgrounds, this study assessed knowledge, attitudes, and awareness in an ethnically diverse community. METHODS: A telephone survey in English or Spanish of a cross sectional, random sample of 692 non-Hispanic whites (NHWs) and 300 Hispanics in Colorado. RESULTS: For all respondent groups, knowledge of appropriate antibiotic use for colds and bronchitis was low. Hispanics surveyed in Spanish, compared with non-Hispanic whites, had significantly lower knowledge about antibiotics for colds, higher knowledge for bronchitis, lower awareness about antibiotic resistance, and greater dissatisfaction if an antibiotic were not prescribed. In all comparisons, English-language Hispanics tended to reflect non-Hispanic white response patterns. Independent predictors of awareness were ethnicity, education, and age. Independent predictors of dissatisfaction were ethnicity, knowledge about antibiotic use for colds, and bronchitis. Ethnicity was an independent predictor of knowledge about the inappropriateness of antibiotics for colds and bronchitis. CONCLUSIONS: To bridge knowledge gaps, educational campaigns for all segments of the population are needed. Content should be responsive to heterogeneity within populations. PMID- 15533526 TI - The AMA proposal to mandate nicotine reduction in cigarettes: a simulation of the population health impacts. AB - BACKGROUND: The American Medical Association (AMA) has advocated gradually reducing the nicotine content of cigarettes to decrease smoking prevalence. Some experts have voiced concerns that smokers may "compensate" by smoking more cigarettes or inhaling more deeply. Further, a black market may emerge, perpetuating cigarette availability. Thus, it is unclear whether a federal mandate would result in a net increase or decrease in population health. The purpose of this research is to estimate the long-term health gains or losses that are likely to accrue to the US population if the nicotine content of cigarettes is gradually reduced to trace levels over a 6-year period. METHODS: To estimate health impacts, we created the Tobacco Policy Model, a computer simulation model. The model simulates the US population as they age and change their smoking behavior over time. Secondary data for model parameters were obtained from publicly available sources. Population health impacts were measured as the change in cumulative quality-adjusted life-years (QALYs) in the US population over 50 years. RESULTS: Following a mandate to reduce nicotine, smoking prevalence is likely to decline from 23% to 5% of the population. Accordingly, a cumulative gain of 157 million QALYs is expected over 50 years. CONCLUSIONS: Despite any mortality increases due to compensatory smoking or the emergence of a black market, implementation of the AMA proposal would likely prevent the addiction of scores of new smokers and result in important gains to the nation's health. This research should prove useful to Congress as they contemplate giving the FDA the authority to regulate tobacco. PMID- 15533527 TI - Comparison of the factors that influence obesity prevalence in three district municipalities of the same city with different socioeconomical status: a survey analysis in an urban Turkish population. AB - BACKGROUND: To evaluate the obesity status, factors and comorbidities related to it in three district municipalities (DM) that compose city center of Bursa with inhabitants of different socioeconomic status. METHODS: A total of 1632 inhabitants >/=18 years of age were interviewed. The number of sample in each DM was obtained proportional to their populations by stratified sampling method. Among 1632, a total of 1543 subjects were included by random sampling and a questionnaire was filled in including demographic, social and behavioral features. RESULTS: The participants living in DM with the highest socioeconomical status (SES) score and level of education had the lowest body mass index (BMI) and body fat percentage (%BF) compared to other DMs. The lowest obesity prevalence (30.8% vs. 36.4% and 39.3%) in that DM was possibly due to younger age, lower female ratio, more active professional, higher percentage of smoking, more consumption of vegetables, olive or corn oil, and less carbohydrate. For the evaluation of the factors that may influence obesity risk, we investigated the effects of these factors in men and women separately with logistic regression model. Sedentary life style and dyslipidemia (DL) in men, being unemployed, having lower level of education and having hypertension (HT) in women and familial obesity in both gender were found to be related to increased obesity risk. CONCLUSIONS: The prevalence of obesity in Bursa is increasing although inhabitants are taking some precautions parallel to their socioeconomical and educational levels. Obesity is becoming a more alarming public health problem in Bursa and Turkey like in most other parts of the world, which forces us to invent new prevention policies. Besides, the results of our study highlight the fact that especially female education requires more attention for decreasing obesity prevalence in coming generations. PMID- 15533528 TI - Perceptions regarding preventive sexual practices and communication with sexual partners among Chinese college students. AB - PURPOSE: To assess the relation between Chinese college students' perceptions and attitudes regarding preventive sexual behaviors, their sexual knowledge, and their use of protective sexual behaviors. METHODS: A subsample of 133 sexually active college students (18-25 years old) was obtained to identify factors related to protective sexual behaviors (e.g., knowledge of partner's sexual history, condom use). RESULTS: Seventy-five percent of the sexually active students were male. Significantly more females had discussed AIDS and sexually transmitted diseases (STDs) issues with their partners and had asked their partners about their sexual histories than males. Students endorsing the use of preventive behaviors were more likely to have discussed AIDS issues with their partners, used condoms, and known their partner's sexual history. CONCLUSIONS: Student perceptions about preventive behaviors have a significant impact on their knowledge of their sexual partners and their own sexual behaviors. These findings, as well as the presence of significant gender differences in perceptions and communications with their partners, have major implications for risk reduction programs. PMID- 15533529 TI - Can government policies help adolescents avoid risky behavior? AB - BACKGROUND: This study examines the extent to which policies influence participation of adolescents in alcohol and tobacco consumption and in unsafe sex. METHODS: Data were obtained from the 1995 Youth Risk Behavior Surveys (YRBS) conducted by 20 different states and cities in the U.S. These data were combined with state data on cigarette taxes, vending machine laws, beer taxes, and family planning clinic availability. A model of teenage risk taking suggested that the three risk behaviors were codetermined by a common latent risk-taking propensity. We used a structural equation model (SEM) accounting for this shared latent propensity to estimate the extent of participation in terms of frequency of smoking, drinking, and the number of sex partners. RESULTS: Estimating simultaneous equations for all three risk behaviors was statistically more efficient than equation-by-equation estimates of each behavior. Estimates indicated significant deterrent effects of beer taxes, vending machine restrictions, and increased density of family planning clinics on teenage risk behavior. CONCLUSIONS: State policies, such as taxes on beer, and restrictions on location of cigarette vending machines, and placement of family planning clinics influence adolescents' behavior. Because there is interrelationship between these behaviors, systems estimators, can offer improved estimates of these effects. PMID- 15533530 TI - The relationship of weight-related perceptions, goals, and behaviors with fruit and vegetable consumption in young adolescents. AB - BACKGROUND: The effect of dieting and weight-control perceptions, goals, and behaviors on food intake, including fruit and vegetable (F&V) consumption, is not completely understood. We examined these associations in 1,755 young adolescents. METHODS: Data from the Teens Eating for Energy and Nutrition at School (TEENS) study were analyzed separately by gender using mixed linear modeling. Surveys completed at the beginning and end of 7th grade asked students to self-report fruit and vegetable consumption, weight-related perceptions, goals, and behaviors, and demographic variables. RESULTS: Underweight boys reported 0.8 fewer F&V servings per day than boys who reported their weight to be 'about right' (P = 0.002), while overweight girls reported 0.7 fewer F&V than girls who reported their weight to be 'about right' (P = 0.005). Boys with weight-related goals consumed significantly more F&V than did boys not wanting to do anything about their weight (P < 0.05). Girls engaging in weight-related behaviors reported 0.7 more F&V than girls who did not engage in weight-related behaviors (P = 0.006). CONCLUSIONS: Because F&V intake is low among all adolescents, population interventions should take place to increase F&V intake. These data support the inclusion of gender-specific intervention components related to weight-related perceptions, goals, and behaviors. PMID- 15533531 TI - Getting focused: missed opportunities for smoking interventions for pregnant women receiving Medicaid. AB - BACKGROUND: The prevalence of smoking, and cessation and relapse rates for pregnant women have health and financial implications. Our objectives were to describe smoking among pregnant smokers receiving Medicaid including characteristics associated with reporting discussion of smoking with providers and the association between those discussions with quitting and maintenance. METHODS: Analysis of Pregnancy Risk Assessment Monitoring System (PRAMS) data from 15 states for 20,287 women with Medicaid for prenatal care during 1998-2000. RESULTS: Thirty-four percent of women smoked before pregnancy (N = 7,686). Most smokers (93%) and nonsmokers (88%) reported discussions about smoking during prenatal care. Women were less likely to have discussed smoking if they were lighter smokers (OR = 1.47; CI = 1.03, 2.12), or reported a previous low birthweight infant (OR = 1.72; CI = 1.03-2.86). Women reporting discussions (compared to those not) were less likely to quit (ARR = 0.70: CI = 0.59-0.91). Quitters reporting discussions (compared to those not) were no more likely to maintain cessation (ARR = 0.89; CI = 0.7, 1.21). CONCLUSIONS: Smoking cessation interventions can be improved for pregnant women receiving Medicaid, especially if focused to address individual needs of light smokers, those with previous low birthweight infants, or those who find it most difficult to quit. PMID- 15533532 TI - The superiority of waist-to-height ratio as an anthropometric index to evaluate clustering of coronary risk factors among non-obese men and women. AB - BACKGROUND: Overtly obesity is relatively rare among the Japanese despite the high prevalence of metabolic disorders, which suggests the need to develop simple and effective methods for assessing metabolic risks among the non-obese individuals as part of public health education. METHODS: We compared body mass index (BMI), waist circumference, and waist-to-height ratio (W/Ht) as indices for evaluation of clustering of coronary risk factors (hypertension, hyperglycemia, hypertriglyceridemia, hypercholesterolemia, and low HDL cholesterol) in 4,668 men and 1,853 women with BMI < 25 kg/m(2). RESULTS: The sum of coronary risk factors correlated positively with all anthropometric indices, with the closest correlation found for W/Ht. Multiple regression analysis showed that height was a negative independent predictor of the sum of coronary risk factors, while age and waist circumference were positive independent predictors. Among the various proposed anthropometric indices for the evaluation of metabolic risk, the sensitivities for identification of clustering of >/=2 and >/=3 coronary risk factors were highest for a waist-to-height ratio >/=0.5 in both genders. CONCLUSIONS: Waist-to-height ratio is more sensitive than BMI or waist circumference alone to evaluate clustering of coronary risk factors among non obese men and women. PMID- 15533533 TI - Adolescents' intentions to smoke as a predictor of smoking. AB - BACKGROUND: Adolescents' intentions to smoke are generally regarded as a valid and reliable predictor of subsequent smoking. This association is largely based on research with adults and needs a more detailed analysis for adolescents. METHODS: Data on intentions and smoking status were collected as part of a longitudinal, birth-cohort study when the study members were 9, 11, 13, 15, 18, and 21 years of age. RESULTS: The results showed that intention to smoke only had an important predictive power in the subgroup of previous nonsmokers. Among those already smoking (on a monthly basis or greater), previous level of smoking was a more important predictor of future behavior than intention to smoke. In addition, the effect of positive intention to smoke was nonlinear over age and had the greatest effect at age 15. CONCLUSION: The results indicated that in adolescence, measurement of intentions to smoke or not smoke cannot be assumed to be a general predictor of behavior at a later age for all groups of adolescents. PMID- 15533534 TI - Back care programs for health promotion-representative user profiles and correlates of participation in Germany. AB - BACKGROUND: In response to the growing incidence of back pain, there is an increasing emphasis on individualized preventive measures such as back care programs. The purpose of the present study was to investigate representative participation rates and their correlates in the German general population. METHODS: The first National Health Survey was carried out in the Federal Republic of Germany from the period of October 1997 to March 1999. It comprised a representative, cross-sectional study of the population with a total sample of 6,159 participants between the ages of 18 and 79. RESULTS: Approximately one in six Germans (16%) has ever participated in a back care program. For 6% of the total, participation took place within the past year. After making adjustments for subjective back pain in the multiple analyses, the 1-year participation rate was significantly lower in men, full-time-workers, one-member households, and those with an unhealthy lifestyle (physically inactive subjects with an unhealthy diet and poorly developed preventive behavior patterns). CONCLUSIONS: The user profile for back care programs on a nationwide scale indicates that genuine needs are not being met and the target group is not being reached. A "preaching to the converted" phenomenon is evident, as indicated by the fact that the user group is made up of individuals who already have a healthy lifestyle and few risks, while the population group most likely to develop back pain is significantly less likely to participate in preventive back care programs. PMID- 15533535 TI - Local school district adoption of state-recommended policies on HIV prevention education. AB - BACKGROUND: This study evaluated the extent to which school districts in Massachusetts adopted HIV education policies consistent with state education agency recommendations, and whether adoption of state-recommended policy language was associated with other core components of school-based HIV prevention programs such as staff development, curriculum, and implementation characteristics. METHODS: A census of health coordinators (n = 251) and high school HIV teachers (n = 174) in randomly selected schools in Massachusetts were surveyed. Chi squares and analysis of variance (ANOVAs) were used to analyze data. RESULTS: Most districts' policies fully incorporated state-recommended language for training HIV teachers (62%), providing HIV education within comprehensive sexuality education (62%), and providing skills-based instruction (57%). Districts adopting state-recommended policies were significantly more likely to have trained more HIV teachers (82% vs. 59% of teachers trained; P < 0.001), provided HIV education to a greater percentage of students (90% vs. 50% of students educated; P < 0.001), and adopted research-based curricula (44% vs. 27%; P < 0.01). High school teachers who received training and those using research based curricula covered more HIV prevention topics and used more skills-based instructional methods than those who did not receive training or did not use research-based curricula (P < 0.01). CONCLUSIONS: Results suggest that strong, state-level HIV prevention education policy recommendations can help shape local school health policy and, when adopted locally, can positively influence the reach and quality of HIV education. PMID- 15533536 TI - The TEAM project: the effectiveness of smoking cessation intervention with hospital patients. AB - BACKGROUND: This study evaluated the effectiveness of three smoking cessation interventions for this population: (1) modified usual care (UC); (2) brief advice (A); and (3) brief advice plus more extended counseling during and after hospitalization (A + C). METHODS: Smokers (2,095) who were in-patients in four hospitals were randomly assigned to condition. Smoking status was ascertained via phone interview 7 days and 12 months post-discharge. At 12 months, reports of abstinence were validated by analysis of saliva cotinine. Intent to treat analyses were performed. RESULTS: At 7-day follow-up, 24.2% of participants reported abstinence in the previous 7 days. There were no differences between conditions. At 12-month follow-up, self-reported abstinence was significantly higher in the A + C condition (UC (15.0%) vs. A (15.2%) vs. A + C (19.8%)). There was no significant difference among conditions in cotinine-validated abstinence, however (UC (8.8%) vs. A (10.0%) vs. A + C (9.9%)). CONCLUSIONS: These interventions for hospital in-patients did not increase abstinence rates. Features of the study that might have contributed to this finding were the inclusiveness of the participation criteria, the fact that pharmacological aids were not provided, and a stage-matching approach that resulted in less intensive counseling for participants unwilling to set a quit date. PMID- 15533537 TI - Evaluation of a multicomponent worksite health promotion program for cardiovascular risk factors-correcting for the regression towards the mean effect. AB - BACKGROUND: We assessed the impact of a multicomponent worksite health promotion program for0 reducing cardiovascular risk factors (CVRF) with short intervention, adjusting for regression towards the mean (RTM) affecting such nonexperimental study without control group. METHODS: A cohort of 4,198 workers (aged 42 +/- 10 years, range 16-76 years, 27% women) were analyzed at 3.7-year interval and stratified by each CVRF risk category (low/medium/high blood pressure [BP], total cholesterol [TC], body mass index [BMI], and smoking) with RTM and secular trend adjustments. Intervention consisted of 15 min CVRF screening and individualized counseling by health professionals to medium- and high-risk individuals, with eventual physician referral. RESULTS: High-risk groups participants improved diastolic BP (-3.4 mm Hg [95%CI: -5.1, -1.7]) in 190 hypertensive patients, TC ( 0.58 mmol/l [-0.71, -0.44]) in 693 hypercholesterolemic patients, and smoking ( 3.1 cig/day [-3.9, -2.3]) in 808 smokers, while systolic BP changes reflected RTM. Low-risk individuals without counseling deteriorated TC and BMI. Body weight increased uniformly in all risk groups (+0.35 kg/year). CONCLUSIONS: In real world conditions, short intervention program participants in high-risk groups for diastolic BP, TC, and smoking improved their CVRF, whereas low-risk TC and BMI groups deteriorated. Future programs may include specific advises to low-risk groups to maintain a favorable CVRF profile. PMID- 15533538 TI - Skin self-examination practices in a convenience sample of U.S. university students. AB - BACKGROUND: Melanoma skin cancer affects many young adults, yet few practice skin self-examination (SSE). We collected detailed information about young adult SSE practices, which can be used to guide the development of SSE interventions that target this age group. METHODS: We surveyed 190 US university students to assess their SSE practices, including thoroughness of self-exams and reasons for not performing SSE. RESULTS: Just 33.2% of respondents had ever performed SSE, and only 5.8% had checked their entire body. The three most commonly cited reasons for failing to practice SSE were not knowing what to look for (55.9%), never thinking of it (54.3%), and not knowing it should be done (33.1%). CONCLUSIONS: Interventions to promote early melanoma detection must raise awareness about the importance of beginning SSE practice in young adulthood and conducting thorough self-exams. They should target all young adults, including those who already practice SSE. PMID- 15533539 TI - Global initiative for chronic obstructive lung disease for chronic obstructive pulmonary disease: GOLD opportunity for lung disorders. AB - BACKGROUND: The purpose of this study was to assess the agreement of asthma and chronic obstructive pulmonary disease (COPD) treatment prescribed by physicians and pulmonologists in comparison to asthma and COPD guidelines and the need of the implementation of COPD guidelines in primary health care physicians. METHODS: Eighty-three asthma patients and 100 COPD patients were chosen and classified in relation to the agreement of their treatment prescribed by a health care physician and that mentioned by global initiative for chronic obstructive lung disease (GOLD) The COPD patients were classified according to their stage of the disease too. RESULTS: Both pulmonologists and primary health care physicians manage asthma patients following asthma guidelines, while great proportion of COPD patients are undertreated by primary health care physicians. The proportion of undertreated COPD patients is decreased as the stage of disease is progressing. CONCLUSIONS: COPD patients mainly are undertreated by primary health care physicians when they are in the primary stages of the disease. The overtreatment of some patients consists of high doses of inhaled steroids prescribed by both pulmonologists, and mainly, primary health care physicians. Therefore, it can be concluded that there is the need of the implementation of COPD guideline by primary health care physicians and the need of COPD patients to be diagnosed in early stages by performing spirometry. PMID- 15533540 TI - It is possible to help smokers in early motivational stages to quit. The Inter99 study. AB - BACKGROUND: In a population-based sample of smokers in early motivational stages, we found a high acceptance of smoking cessation groups. METHODS: Inter99 is a randomized population-based intervention study, in Copenhagen, Denmark. Smokers in all motivational stages were included. All participants underwent a lifestyle consultation and 2,168 smokers in the high intensity intervention group were offered assistance to quit in smoking cessation groups. RESULTS: Thirty-five percent were validated to be continuously abstinent at the end of the smoking cessation groups. Eighty-four percent of the smokers achieving sustained abstinence in our study had no serious plans to quit soon before the lifestyle consultation. Motivation to quit before the lifestyle consultation could not predict abstinence. Being a man, and having a job and at least 1 year of vocational training were predictors of abstinence in a multivariate model, whereas high nicotine dependence and living with a smoking spouse were predictors of failure. CONCLUSION: High cessation rates were obtained in a population of heavy smokers with moderate nicotine dependence. It was possible to obtain sustained abstinence in smokers in early motivational stages. These smokers would probably not have been reached by traditional smoking campaigns. PMID- 15533541 TI - Smoking cessation intervention in a large randomised population-based study. The Inter99 study. AB - BACKGROUND: Several large and well-conducted community interventions have failed to detect an effect on prevalence of smoking. METHODS: Two thousand four hundred eight daily smokers in all motivational stages were actively recruited and included in a randomised population-based intervention study in Copenhagen, Denmark. All smokers completed a questionnaire and underwent a health examination and a lifestyle consultation. Daily smokers in the high intensity intervention group were offered assistance to quit in smoking cessation groups. RESULTS: The validated abstinence rate at 1-year follow-up was 16.3% in the high intensity group and 12.7% in the low intensity group compared with a self-reported abstinence rate of 7.3% in the background population. The adjusted odds ratio of abstinence in the high intervention group was significantly higher, OR = 2.2 (1.6 3.0) than in the background population, also in the 'intention-to-treat' analyses, OR = 1.5 (1.1-2.0). Higher socioeconomic status, higher age at onset of daily smoking, and a higher wish to quit were predictors of success. CONCLUSION: In a population-based setting, using active recruitment and offering assistance to quit, it was possible to include many smokers and to achieve a significantly higher validated abstinence in the high intensity intervention than in the background population, even when using 'intention-to-treat' analyses. PMID- 15533542 TI - How many days of pedometer monitoring predict weekly physical activity in adults? AB - BACKGROUND: The study purpose was to establish the number (and type) of days needed to estimate mean pedometer-determined steps/day in a field setting. METHODS: Seven days of data were collected from 90 participants (33 males, age = 49.1 +/- 16.2 years, BMI = 27.2 +/- 4.1 kg/m(2); 57 females, age = 44.8 +/- 16.9 years, BMI = 27.0 +/- 5.9 kg/m(2)). Mean steps/day were computed for all 7 days (the criterion), each single day, and combinations of days. Analyses included repeated measures ANOVA, intra-class correlations (ICC), and regression. RESULTS: There was a significant difference (P < 0.001) between days. The difference was limited to Sunday and accounted for 5% of the variance. ICC analyses indicated a minimum of 3 days is necessary to achieve a reliability of 0.80. The adjusted R(2) was 0.79 for a single day (specifically Wednesday), 0.89 for 2 days (Wednesday, Thursday), and 0.94 for 3 days (Wednesday, Thursday, Friday). Sunday was the last day to enter the model. CONCLUSIONS: Although there is a statistical difference between days, there is little practical difference, and the primary distinction appears limited to Sunday. Although a single day of collection is not acceptable, any 3 days can provide a sufficient estimate. PMID- 15533543 TI - Mediterranean diet and cardiovascular diseases in an Israeli population. AB - BACKGROUND: Israel is in the Mediterranean basin, but its population immigrated mostly from Europe, Asia, and Africa. We surveyed the adherence of the Israeli Jewish population to Mediterranean Diet (MD) and its association with cardiovascular disease. METHODS: We studied a random sample of Jewish adults, aged 35+ using a 24-h recall questionnaire. A MD score (scale 0-8) was computed reflecting high ratio of monounsaturated to saturated fat; high intake of alcohol, legumes, cereals, vegetables, and fruits; low intakes of meat and it's products, milk and dairy products. Scoring <5 was defined as Low-MD consuming, while 5+ as High-MD consuming. RESULTS: Five hundred twenty men and 639 women participated in the study, with High-MD consumers constituting 19% and 17% by sex, respectively. The risk for myocardial infarction, coronary bypass, angioplasty, and any cardiovascular disease in men increased by 1.2 (P = 0.04), 1.6 (P = 0.01), 1.4 (P = 0.003), and 1.3 (P = 0.01), respectively, for each MD score decrease. In women, crude odds ratios ranged from 1.4 to 1.9 but were not statistically significant. CONCLUSIONS: The current rate of MD in Israel is fairly low. Given the MD's proven preventive effect and local availability of foods, a reasonably priced MD diet can be devised and recommended widely in our population. PMID- 15533544 TI - Does extended proactive telephone support increase smoking cessation among low income women using nicotine patches? AB - BACKGROUND: It is unclear whether proactive telephone support enhances smoking cessation beyond the provision of nicotine replacement therapy alone. METHODS: We randomly assigned 330 low-income women smokers to receive either free nicotine patches (control condition) or free nicotine patches with up to 16 weeks of proactive telephone support (experimental condition). All participants were assessed by telephone at baseline and at 2 weeks, 3 months, and 6 months post baseline to determine smoking status. RESULTS: Results revealed a significant effect for the telephone support at 3 months, with 43% of experimental versus 26% of control condition women reporting 30-day point prevalent abstinence (P = 0.002). The difference was no longer significant at 6 months. A metaanalysis conducted with five randomized studies revealed a slight but non-significant long term benefit of proactive telephone support when added to the provision of free nicotine patches for smoking cessation. CONCLUSIONS: This is the second study to demonstrate a short-term effect for proactive telephone support added to free nicotine replacement therapy; however, neither the current study, nor the metaanalysis including the four other published trials, confirmed a longer-term benefit. PMID- 15533545 TI - Does the effect of behavioral counseling on fruit and vegetable intake vary with stage of readiness to change? AB - BACKGROUND: We have recently shown that brief behavioral counseling based on the stage of change (SOC) model stimulates greater increases in fruit and vegetable intake over 12 months than nutritional education in adults living in a low-income urban area. We tested the hypothesis that behavioral counseling would overcome the greater obstacles to change in precontemplators and contemplators compared with those initially in the preparation stage. METHOD: Two hundred and seventy one adults took part in a parallel group randomised controlled trial comparing behavioral counseling and nutritional education. Counseling was delivered in two 15-min sessions and accompanied by written material. Self-report changes in fruit and vegetable consumption over 12 months were analysed on an intention-to-treat basis and related to baseline stage of change. RESULTS: At baseline, 148 (54.6%) of participants were in preparation, 54 (19.9%) in contemplation and 69 (25.5%) in precontemplation. Preparers were younger, more educated and more likely to be female than were precontemplators and contemplators. In the nutritional education group, baseline stage predicted changes over 12 months, with larger increases in fruits and vegetables in the preparation than in the precontemplation or contemplation groups. This was not the case with behavioral counseling, in which increases in consumption were unrelated to baseline stage. CONCLUSION: Tailored behavioral counseling helped to overcome the barriers to increasing fruit and vegetable intake present among participants in contemplation stage but not the precontemplation or preparation stages. PMID- 15533546 TI - Lipid profile in spinal cord-injured women with different injury levels. AB - Background. In dependence on their injury level, male subjects with spinal cord injury (SCI) exhibit a less favorable lipoprotein profile than control persons. The impairment of the sympathetic nervous system and the fact that persons with spinal cord injury are subject to extreme physical inactivity may have an influence on their lipid profile and lipoprotein(a) concentration. It has been shown that sex-specific differences in hormonal regulation are responsible for differences in lipoprotein levels between nondisabled men and women. However, the role of hormones on lipoprotein levels has not been investigated in female subjects with spinal cord injury. Methods. Therefore, we performed a detailed investigation regarding the lipid profile in 32 premenopausal women with spinal cord injury ranging from tetraplegia to low paraplegia and in 36 control subjects. VO(2max) was determined by a wheelchair ergometry with stepwise increase in work load. Result. VO(2max) was significantly higher in paraplegics than in tetraplegics but significantly lower than in control subjects. Paraplegics had significantly higher low-density lipoprotein levels than both tetraplegics and control persons. The lipid profile of female tetraplegics was characterized by elevated triglycerides. An association between high-density lipoprotein levels and spinal cord injury or the level of the injury was not observed. No significant difference in lipoprotein(a) was found within SCI individuals as well as between SCI individuals and control persons indicating the predominant genetic determination of lipoprotein(a) and the thus related cardiovascular risk. Conclusion. Despite the extreme reduction of VO(2max), the assumed physical inactivity and low serum catecholamine levels due to the impairment of the sympathetic nervous system, female tetraplegic persons did not show an adverse lipoprotein profile with respect to high-density lipoprotein cholesterol levels. If the higher low-density lipoprotein cholesterol concentrations in female with spinal cord injury with low lesion levels or the elevated TG levels in female tetraplegics bare relevance with respect to an increased cardiovascular risk in this population needs to be clarified in further longitudinal investigations. PMID- 15533547 TI - Relationship of cognitive restraint of eating and disinhibition to the quality of food choices of Latina women and their young children. AB - BACKGROUND: Previous studies have examined the association of methods to control eating such as dietary restraint and disinhibition with weight and quantity of foods eaten. The purpose of this study was to examine the association of these constructs with quality of food choices for women and for their young children. METHODS: One hundred and eighty-seven Latina women completed the Eating Inventory, which measures cognitive dietary restraint and disinhibition, and reported on the food intakes of themselves and their 5- to 7-year-old child using a food frequency questionnaire. BMIs were also calculated. RESULTS: Cognitive restraint in mothers was generally associated with more healthful food choices of themselves and, to a lesser degree, of their children. Mothers' dietary disinhibition was associated with less healthful choices of themselves and their children. Mothers' control variables were not related to children's BMIs, except disinhibition was positively correlated with boys' BMI. Cognitive restraint in this low-income Latina population is thus associated with higher quality diets of self and child with no negative impact on children's BMIs whereas disinhibition is associated with overeating by self and child of high-fat, high-sugar foods. With the rise of obesity and with dietary quality of most Americans needing improvement, restraint may be a form of necessary cognitive self-regulation; and disinhibition or overeating is of greater concern. PMID- 15533548 TI - Access to health care among older adults and receipt of preventive services. Results from the Behavioral Risk Factor Surveillance System, 2002. AB - BACKGROUND: Although most of the 34 million U.S. adults aged 65 years and older have health care coverage, many do not receive preventive care. To investigate why, we examined various barriers to access of health care and their effect on obtaining preventive care. METHODS: A cross-sectional study was conducted of noninstitutionalized adults, aged 65 years or older, in states that participated in the Behavioral Risk Factor Surveillance System in 2002. RESULTS: Of the 46,659 respondents aged 65 years and older, 93% had a regular care provider, 98% had a regular place of care, and 98% were able to obtain needed medical care. Those with a regular care provider or a regular place of care were more likely to receive clinical preventive services than those without either of these. Reasons for not obtaining needed medical care were cost (27%), too long a wait for an appointment (20%), no transportation or distance (9%), office not open when the individual could get there (8%), and other reasons (32%). CONCLUSIONS: Having a regular care provider or a regular place of care is associated with a significant likelihood of receipt of clinical preventive services among older adults. Efforts to eliminate barriers to health care access may increase older adults' receipt of such services. PMID- 15533549 TI - A population study of the self-care behaviors and their associated factors of diabetes in Taiwan: results from the 2001 National Health Interview Survey in Taiwan. AB - BACKGROUND: Diabetes has been the fifth leading cause of death in Taiwan since 1987, and the complications are a burden to the patients, their families, and society. PURPOSE: The purpose of this study was to examine the self-control behaviors of diabetics and associated factors. METHODS: Data for this study came from the 2001 National Health Interview Survey in Taiwan. Subjects with diabetes confirmed by medical professionals, aged 40 years or older, were included. Self care behaviors included any of the following: taking medicine regularly, reducing weight, avoiding cigarettes or alcohol, exercising, practicing diet control, and maintaining a regular life style and avoiding late hours. The factors considered were demographic factors including age, sex, education, income, and health behavior factors including smoking, drinking alcohol, chewing betel nuts, and health conditions such as existence of other chronic diseases, limitation of activity of daily living (ADL), and self-perceived health. RESULTS: There were 764 subjects aged 40 years or older who were diagnosed with diabetes. Logistic regression showed that females, those with older age or chronic disease such as hypertension or dyslipidemia, and those with long duration of diabetes took medicine regularly. Old age group, middle school educated, and longer duration of the disease had a higher chance of using any self-care method than their counterparts. CONCLUSION: Attention should be given to the younger age group, shorter duration of the disease in practicing self-care of diabetes. PMID- 15533550 TI - Relationships and CHD risks of BMI, lipoproteins, lipids, and blood pressure in African-American men and women. AB - BACKGROUND: Body mass index (BMI) is believed to be associated with coronary heart disease (CHD) risks. A major question is whether each BMI category provides independent risk information and whether the BMI categories provide similar results for African-American (AA) men and women relative to lipid, lipoproteins, and blood pressure. METHODS: A descriptive research design was used to evaluate the effects of BMI categories on lipoproteins, lipids, and blood pressure, and to determine if similar CHD risks related to these variables were observed in AA men and women. The 358 (108 men and 250 women; 40-70 year of age) participants were recruited from churches in a southeastern metropolitan area. RESULTS: The results indicate that BMI is positively associated with lipoproteins, lipids, and blood pressure in both AA men and women. Relative risks for CHD were higher for obese (BMI > 30 kg/m(2)) men and women, but the overweight (OW) category provided the same information as the normal weight (NW) category for men, but not for women. CONCLUSIONS: This suggests that the same BMI standards provide different CHD risk information for AA men than for AA women and that gender-specific BMI standards may be warranted for more effective CHD risk stratification in AA adults. PMID- 15533551 TI - Community level alcohol availability and enforcement of possession laws as predictors of youth drinking. AB - BACKGROUND: Despite a minimum legal drinking age, many young people use alcohol. Environmental strategies to control youth drinking focus on restricting access and the enforcement of possession laws. This study examines the relationship between use of these strategies and the frequency of youth alcohol use and related problems. METHODS: Participants were 16,694 students, ages 16-17 in 92 communities in Oregon. A multi-level analysis of a repeated cross-sectional statewide student survey was conducted. The outcome measures examined include 30 day frequency of alcohol use, binge drinking, use of alcohol at school, and drinking and driving. RESULTS: The rate of illegal merchant sales in the communities directly related to all four alcohol-use outcomes. There was also evidence that communities with higher minor in possession law enforcement had lower rates of alcohol use and binge drinking. The use of various sources in a community expanded and contracted somewhat depending on levels of access and enforcement. CONCLUSIONS: This evidence provides empirical support for the potential utility of local efforts to maintain or increase alcohol access control and possession enforcement. PMID- 15533552 TI - Limitations of the dual-process-theory regarding the writing of words and non words to dictation. AB - It is generally assumed that the lexical and phonological systems are involved in writing to dictation. In an experiment concerned with the writing of words and non-words to dictation, the handwriting of female students was registered using a digitising tablet. The data contradict the assumption that the phonological system represents an alexical process. Both words and non-words which were acoustically presented to the subjects were lexically parsed. The analysis of kinematic data revealed significant differences between the subjects' writing of words and non-words. The findings reveal gross disturbances of handwriting fluency during the writing of non-words. The findings of the experiment cannot be explained by the dual-process-theory. PMID- 15533553 TI - Normative data for the Boston Naming Test in native Dutch-speaking Belgian children and the relation with intelligence. AB - This paper reports the results of a normative study of the 60-item version of the Boston Naming Test (BNT) in a group of 371 native Dutch-speaking Flemish children between the ages of 6 and 12 years. Analysis of test results revealed that BNT performance was significantly affected by age and gender. The gathered norms were shown to be significantly lower than published norms for comparable North American children. Error analysis disclosed remarkable similarities with data from elderly subjects, with verbal semantic paraphasias and 'don't know' responses occurring most frequently. Finally, BNT scores were shown to correlate strongly with general intelligence as measured with the Raven Progressive Matrices. The relation between both measures can be of help in the diagnosis of identification naming deficits and impaired word-retrieval capacities. PMID- 15533554 TI - Evidence for separate tonal and segmental tiers in the lexical specification of words: a case study of a brain-damaged Chinese speaker. AB - We present an acoustic study of segmental and prosodic properties of words produced by a female speaker of Chinese with left-hemisphere brain damage. We measured the location of the point vowels /a, e, [Symbol: see text], i, y, o, u/ and determined their separation in the vowel plane, and their perceptual distinctivity. Similarly, the acoustic properties of the four lexical tones were measured in the F0 x time space. The data for our brain-damaged speaker were compared with those of a healthy control speaker. Results show that the patient's vowels hardly suffered from her lesion (relative to the vowel dispersion in the healthy control speaker), but that the identifiability of the four lexical tones was greatly compromised. These findings show that the tonal errors in aphasic speech behave independently of the segmental errors, even though both serve to maintain lexical contrasts in Chinese, and are therefore part of the lexical specification of Chinese words. The present study suggests that the specification of segmental and tonal aspects of lexical entries in Chinese, and in tone languages in general, are located or processed separately in the brain. PMID- 15533555 TI - Cognitive and cognate-based treatments for bilingual aphasia: a case study. AB - Two consecutive treatments were conducted to investigate skill learning and generalization within and across cognitive-linguistic domains in a 62-year-old Spanish-English bilingual man with severe non-fluent aphasia. Treatment 1 was a cognitive-based treatment that emphasized non-linguistic skills, such as visual scanning, categorization, and simple arithmetic. Treatment 2 was a lexically based treatment that trained cognates (cross-linguistic word pairs that are similar in meaning and form, such as rosa/rose) and non-cognates (cross linguistic word pairs with shared meaning but different forms, such as mesa/table). Treatment 1 resulted in modest gains in both Spanish and English. Treatment 2 resulted in improved naming for non-cognates as well as cognates within each language. However, the generalization of gains from Spanish to English was apparent only for cognate stimuli. PMID- 15533556 TI - Reading comprehension and understanding idiomatic expressions: a developmental study. AB - The aim of the present study was to investigate idiom comprehension in school-age Italian children with different reading comprehension skills. According to our hypothesis, the level of a child's text comprehension skills should predict his/her ability to understand idiomatic meanings. Idiom comprehension in fact requires children to go beyond a simple word-by-word comprehension strategy and to integrate figurative meaning into contextual information. In a preliminary phase, we used a standardized battery of tests (Cornoldi & Colpo, 1998) to assess the ability of second graders and fourth graders to comprehend written texts. Three groups were identified at each age level: good, medium, and poor comprehenders. Children were then presented with familiar idiomatic expressions which also have a literal meaning (e.g., "break the ice"). Idioms were embedded in short stories: in Experiment 1 only the idiomatic interpretation was contextually appropriate, in Experiment 2 a literal reading of the string was also plausible in the context. A multiple-choice task was used in both experiments: children were asked to choose one answer among three corresponding to: (a) the idiomatic meaning; (b) the literal meaning; and (c) an interpretation contextually appropriate but not connected with the idiomatic or literal meaning of the idiom string. The results of both experiments showed that the ability to understand a text indeed predicted children's understanding of idioms in context. To verify whether possible improvements in children's comprehension skills might produce an increase in figurative language understanding, Experiment 3 was carried out. A group of poor comprehenders who participated in Experiments 1 and 2 was tested eight months later. The results of Experiment 3 showed that children whose general comprehension skills improved their performance on an idiom comprehension test. PMID- 15533557 TI - Dissociable patterns of brain activity during comprehension of rapid and syntactically complex speech: evidence from fMRI. AB - Sentence comprehension is a complex task that involves both language-specific processing components and general cognitive resources. Comprehension can be made more difficult by increasing the syntactic complexity or the presentation rate of a sentence, but it is unclear whether the same neural mechanism underlies both of these effects. In the current study, we used event-related functional magnetic resonance imaging (fMRI) to monitor neural activity while participants heard sentences containing a subject-relative or object-relative center-embedded clause presented at three different speech rates. Syntactically complex object-relative sentences activated left inferior frontal cortex across presentation rates, whereas sentences presented at a rapid rate recruited frontal brain regions such as anterior cingulate and premotor cortex, regardless of syntactic complexity. These results suggest that dissociable components of a large-scale neural network support the processing of syntactic complexity and speech presented at a rapid rate during auditory sentence processing. PMID- 15533558 TI - The effect of script on poor readers' sensitivity to dynamic visual stimuli. AB - The current research examined performance of good and poor readers of Thai on two tasks that assess sensitivity to dynamic visual displays. Readers of Thai, a complex alphabetic script that nonetheless has a regular orthography, were chosen in order to contrast patterns of performance with readers of Korean Hangul (a similarly regular language but one that has a simple visual format, see ). Thai poor readers were less sensitive than good readers on the two measures of dynamic visual processing; they had higher thresholds for detecting coherent movement and required longer ISIs to report group movement in the Ternus task. These results differ from those for poor readers of Korean Hangul that found no relationship between visual processing thresholds and reading skill. This contrast suggests that the expression of visual processing problems in dyslexia is mediated by the format properties of the writing system and points to the need to consider such factors in formulating brain-behavior relationships. PMID- 15533559 TI - Pediatric middle ear infections and gastroesophageal reflux. AB - OBJECTIVES: The pathogenesis of middle ear infections in children is multifactorial and includes infection, anatomical factors, impaired immunologic status, allergy, familial predisposition, male sex, method of feeding and environmental factors. Glue ear remains the commonest cause of deafness in childhood. Gastroesophageal reflux (GOR) is a common problem in the newborn and preschool periods. Recent research suggests that it may be related to eustachian tube dysfunction and otitis media. METHODS: We review the literature and discuss the possible relationship between Gastroesophageal reflux and otitis media in children. CONCLUSIONS: The current data are not enough to support antireflux treatment in children with refractory middle ear infections. More prospective randomised placebo-controlled studies are needed. PMID- 15533560 TI - Analysis of a staging assessment system for prediction of surgical interval in recurrent respiratory papillomatosis. AB - INTRODUCTION: A staging system for assessment of recurrent respiratory papillomatosis (RRP) has been in use over the last 3 years for 17 patients at our children's hospital. OBJECTIVE: To evaluate a staging system for assessment of RRP on the basis of the predictive value on the surgical interval of: anatomic staging system score, urgency of intervention, voice quality, and stridor. To assess effect on surgical interval of the age of patient at time of surgery and use of adjuvant therapy. To develop a model based on the staging criteria to predict need for adjuvant therapy. DESIGN: Validation cohort. SETTING: Academic children's hospital. RESULTS: Estimated time to surgery decreased by "x days": "independent variable" "(95% CI)" as follows. Four days: each 1 point in total anatomical score (0.2, 8); 120 days: total anatomical score >20 (37,203); 143 days: urgent versus scheduled surgery (42, 243); 100 days: aphonic versus normal voice (-211, 343); 31 days: abnormal versus normal voice (-281, 343); 125 days: stridor at rest versus no stridor (-31, 281); 109 days: stridor with activity versus no stridor (19, 198); 26 days: each 1 year decrease in age (22, 30). Adjuvant therapy delays next surgery by 32 days (-4, 69) and average scores decreased while on adjuvant therapy. CONCLUSIONS: Elements of our proposed staging system are effective in prediction of surgical interval with statistical significance achieved for total anatomical scores, urgency of intervention and stridor with activity versus no stridor. This study is pilot in nature and provides a framework upon which future studies can be based. The analysis of a larger, more severity diverse population could yield results which lead to a model capable of effectively predicting future surgical interval based on a weighted prediction score calculated from age, comorbidities, anatomic staging score, and clinical staging score. PMID- 15533561 TI - Surgery for speech in cleft palate patients. AB - INTRODUCTION: Superiorly based pharyngeal flaps and sphincter pharyngoplasties are the two main possibilities for the surgical treatment of hypernasality in velopharyngeal dysfunction. Videonaspharyngoscopy and multi-view videofluoroscopy can provide anatomical and physiological data for planning these surgical procedures for correcting hypernasality. AIM: This study was undertaken to assess the planning and outcome of pharyngeal flaps and sphincter pharyngoplasties for correcting velopharyngeal insufficiency. The surgical techniques were customized according to the findings of videonasopharyngoscopy and multiview videofluoroscopy. MATERIALS AND METHODS: Seventy patients with repaired palate clefts and residual velopharyngeal dysfunction were studied. The patients were randomly divided into two groups. The first group received a pharyngeal flap. The second group received a sphincter pharyngoplasty. Both procedures were individually customized according to the findings of videonasopharyngoscopy and multi-view videofluoroscopy. RESULTS: There was a non-significant difference (P >0.05) between the mean size of preoperative velopharyngeal closure gap between the two groups of patients (mean=27.5%; S.D.=7.7% versus mean=28.3%; S.D.=5.9%). Postoperatively, velopharyngeal dysfunction was completely corrected in 89% of the cases from group 1, and in 85% of the cases from group 2. There was a non significant difference (P >0.05) between the success rate for correcting VPI in both groups of patients. CONCLUSIONS: Pharyngeal flap and sphincter pharyngoplasty seem to be safe and reliable procedures for treating residual velopharyngeal dysfunction. Although not all the patients studied for this paper achieved complete closure after the surgical procedures, all of them showed a reduction of the size of the velopharyngeal closure defect. The planning of the surgical procedure, in order to match the postoperative structure to the preoperative velopharyngeal dimensions and movements visualized through Videonasopharyngoscopy and videofluoroscopy, seems to be the most important aspect of the surgery for correcting residual velopharyngeal dysfunction. PMID- 15533562 TI - Complications in paediatric airway management. AB - Management of the paediatric airway can be difficult. This management in turn can be associated with complications which can be life threatening. We present three cases in which such complications occurred and then go on to discuss the different ways in which these can be prevented. PMID- 15533563 TI - Pediatric caustic ingestion and parental cocaine abuse. AB - OBJECTIVE: To report two cases in which toddlers drank a caustic liquid which their parents had been using to prepare free-base and crack cocaine. DESIGN: Case series. RESULTS: Two toddlers presented with first- and second-degree burns of the oral cavity, pharynx and esophagus from caustic ingestion. The first had drunk ammonia from an unmarked clear plastic container the morning after "a party at a friend's house". The second had drunk potassium hydroxide from a clear plastic container which was labelled "pH UP" and had a small "Danger" sign. Both children were treated with intravenous hydration, steroids, antibiotics and analgesics. Fortunately, neither child suffered long-term physical sequelae. When questioned, the parents of both children admitted to using these alkalis to prepare free-base and crack cocaine. Appropriate steps were taken by social services to protect these children from further neglect. DISCUSSION: Despite federal legislation and public education, pediatric caustic ingestion remains a significant problem. These cases suggest that, in certain cases of pediatric caustic ingestion, caregivers should be questioned regarding possible drug abuse. Larger warning labels and further public education might help prevent many cases of pediatric caustic ingestion. PMID- 15533564 TI - Safety of electronic molecular resonance adenoidectomy. AB - Adenoidectomy is one of the most frequent operations in children. In order to reduce hospital stay costs, today, this procedure is performed as day surgery. Even though adenoidectomy is not considered risky, some minor complications may occur, the most important being bleeding (0.5-8% incidence). The surgical technique used can influence considerably postoperative pain and time to recovery. This aspect is essential in the management of day surgery patients, for whom the need of safe and rapid maneuvers associated with early recovery determines the choice of the surgical procedure. Recently, we developed a surgical technique based on the use of an electronic molecular resonance tool associated with bendable suction electrocautery. This study was carried out on 600 patients, divided into two groups, the first undergoing ablation using the molecular resonance tool and the second undergoing curette adenoidectomy. The two groups were homogeneous for age, sex, surgical indications, and grade of adenoid hypertrophy. The following parameters were considered: duration of surgery, importance of intraoperative bleeding, time to cicatrization, incidence of bleeding complications. Duration of surgery and intra- and postoperative bleeding were much lower in the first group than in the group undergoing traditional adenoidectomy. In addition, rhinopharyngeal complete cicatrization, defined as absolute absence of pseudomembrane, was much quicker in the first group, as assessed by postoperative endoscopy. Another major advantage offered by the molecular resonance tool is the accuracy of surgery performed under visual control in a practically bloodless field. To sum up, this method, thanks to its technical features and safety, is particularly indicated in children and in patients with coagulation disorders. PMID- 15533565 TI - Multi-lobulated cervical pneumatocoele communicating with pyopneumothorax and bronchopleural fistula. AB - Cervical herniation of lung in infants is a very rare condition which may present as a diagnostic dilemma. Reported here is an infant who presented with a swelling apparently looking like an external laryngocoele but was finally diagnosed as an apical pneumatocoele communicating with pyopneumothorax. PMID- 15533566 TI - Recurrent periorbital cellulitis in a child. A random event or an underlying anatomical abnormality? AB - We present a case of recurrent periorbital cellulitis in a child. From the age of 2 years the child was admitted with periorbital cellulitis a total of six times with the first five episodes responding to conservative management with intravenous antibiotics. On the sixth admission endoscopic surgical treatment was carried out. Two abnormalities were noted: an anatomically abnormal uncinate process and an isolated abscess in an ethmoidal cell. It is believed that the abnormal uncinate process caused obstruction of the osteomeatal area, predisposing to recurrent peri-orbital cellulitis. We discuss the importance of early imaging and surgery in recurrent periorbital cellulitis. PMID- 15533567 TI - Solitary infantile myofibroma compromising the airway. AB - Infantile myofibromatosis is an uncommon and benign condition presenting in the neonatal period. It is self-limiting disease that may present as a localised or generalised process. Various examples of this entity have been reported in the literature. This report describes a neonate with a rapidly growing oropharyngeal lesion obstructing the airway that had the typical histological features of an infantile myofibroma. This case report highlights that a solitary myofibroma may be incredibly extensive making complete excision impossible and can be particularly challenging to manage in terms of airway stabilisation. PMID- 15533568 TI - Post-traumatic facial artery aneurysm in a child. AB - Significant facial soft tissue injury can result from trauma to the head and neck. The long-term sequelae of such injuries include vascular anomalies, which may present weeks or months following the initial injury. We report here a case of a post-traumatic facial artery aneurysm which developed in a pediatric patient three weeks following a penetrating facial injury during motor vehicle accident (MVA). Although post-traumatic aneurysm of the facial artery is rare, follow-up on head and neck trauma patients with respect to this type of complication is warranted. PMID- 15533569 TI - Lipofibromatosis presenting as a pediatric neck mass. AB - Lipofibromatosis is a recently described, benign neoplasm that presents in the pediatric population [J.F. Fetsch, M. Miettinen, W.B. Laskin, M. Michal, F.M. Enzinger, A clinicopathologic study of 45 pediatric soft tissue tumors with an admixture of adipose tissue and fibroblastic elements, and a proposal for classification as lipofibromatosis, Am. J. Surg. Pathol. 24 (2000) 1491-1500]. It is a rare soft tissue tumor histologically distinct from other fibromatoses such as juvenile fibromatosis, fibrous hamartoma of infancy, calcifying aponeurotic fibroma, and lipoblastoma. Distinguishing histopathologic features of lipofibromatosis include abundant and disorganized adipose lobules traversed by bundles of spindled fibroblast-like cells. It most commonly presents in the extremities. We present a case involving a young girl, which we believe represents the first report of lipofibromatosis involving the neck. PMID- 15533570 TI - Laryngeal chondroma: a rare neck tumor in pediatric age. AB - Primary chondroid tumors of the larynx constitute less than 1% of all laryngeal tumors and are usually prevalent in adults. Relevant English literature presents only two pediatric laryngeal chondroma cases. In this paper, a novel pediatric case of laryngeal chondroma presenting as an external laryngeal mass is reported. PMID- 15533571 TI - Lipoid proteinosis of larynx: review of four cases. AB - Lipoid proteinosis is a rare autosomal recessive disorder characterized by intercellular deposition of an amorphous hyaline material. It mainly involves skin and mucosal membranes of upper aerodigestive tract as well as central nervous system, lung, lymph nodes and striated muscles. Etiology and pathogenesis are unknown. Infantile hoarseness is a common presenting feature of the disease due to infiltration of larynx. In two-thirds of the cases, voice changes are present at birth or in early infancy as the first manifestation. We present four patients with lipoid proteinosis involving skin, oropharynx and larynx. PMID- 15533572 TI - Balloon dilatation following tracheal reconstruction for congenital microtrachea. AB - A male infant presented with failed extubation was diagnosed with congenital microtrachea. After primary combined surgery with excision of a left pulmonary artery sling and microtracheal resection with anastomosis incorporating the excised microtrachea as an autologous anterior tracheal graft he was treated with balloon dilatation of the re-stenosed segment on three occasions during the following year. During the 4 years follow-up, stridor has been minimal and intermittent. His feeding and growth have been normal. We have found balloon dilatation to be a useful adjunct to tracheal reconstructive surgery in this difficult condition. PMID- 15533573 TI - Obstructive congenital gingival granular cell tumour. AB - Congenital gingival granular cell tumours (CGCT) are rare and always benign intraoral tumours originating from the alveolar ridge. They are also known as congenital epulis, congenital myoblastoma or Neumann's tumour. They are typically seen as a mass protruding out of a newborn child's mouth. In general, CGCT occurs as a solitary tumour. The main differential diagnosis is epignathus (oral teratoma). This report describes a newborn with a mass originating from lower alveolar ridge obtruding into the oral cavity. Clinical features, histiogenesis and necessity for early surgical treatment due to risk of airway obstruction and difficulty in feeding were discussed. PMID- 15533574 TI - Stickler syndrome type I and Stapes ankylosis. AB - OBJECTIVE: To report a successful stapedectomy for stapedial fixation in a patient with Stickler syndrome type I (COL2A1). SETTING: University Hospital Department for Otology, Pathology, Ophthalmology and Clinical Genetics. STUDY DESIGN: A clinical and genetic evaluation of a mother and daughter focusing mainly on the otological, ophthalmological, histological and genetical aspects. INTERVENTION: A stapedectomy was performed successfully. RESULTS: Hearing impairment improved after stapedectomy. Postoperatively a shift in high-frequency threshold wa seen related to the stapedectomy. A new mutation in COL2A1 gene was dectected. CONCLUSION: Stapedial fixation can be the cause of hearing impairment in Stickler syndrome type I (COL2A1). The hearing impairment can be improved by stapes surgery. PMID- 15533575 TI - Prognostic factors of recurrent respiratory papillomatosis spread to lower airway passages. PMID- 15533577 TI - Acute elevation of free fatty acids impairs hepatic glucose uptake in conscious rats. AB - To investigate the dose-dependent effect of free fatty acid (FFA) on the hepatic glucose uptake (HGU), we determined hepatic glucose fluxes by a dual tracer technique during the basal state and euglycemic hyperinsulinemic clamp combined with a portal glucose load in three groups of rats given saline (saline), low dose lipid (lipid-L), or high-dose lipid infusion (lipid-H). In the basal state, lipid infusion dose-dependently increased plasma FFA (saline, 400 +/- 50; lipid L, 550 +/- 30; lipid-H, 1700 +/- 270 micromol l(-1); mean +/- S.E). Endogenous glucose production (EGP) in lipid-H was 63.5 +/- 5.5 micromol kg(-1) min(-1) and significantly higher than in the saline and lipid-L (40.2 +/- 2.9, 47.6 +/- 3.1 micromol kg(-1) min(-1), respectively). During euglycemic hyperinsulinemic clamp, plasma FFA decreased to 130 +/- 30 micromol l(-1) in saline, but remained at basal levels in lipid-L and lipid-H (470 +/- 30 and 1110 +/- 180 micromol l(-1), respectively). Insulin-suppressed EGP was complete in saline and lipid-L, but impaired in lipid-H (38.0 +/- 6.4 micromol kg(-1) min(-1)). Elevated FFA dose dependently reduced HGU (saline, 12.2 +/- 0.9; lipid-L, 8.6 +/- 0.6; lipid-H, 4.7 +/- 1.4 micromol kg(-1) min(-1)). In conclusion, acutely elevated FFA impairs HGU as well as insulin-mediated suppression of EGP during hyperinsulinemic clamp with portal glucose loading. Impaired hepatic glucose uptake associated with elevated FFA may contribute to the development of insulin resistance in obesity and type 2 diabetes. PMID- 15533578 TI - Regulation of fibronectin levels by agmatine and spermine in mesangial cells under high-glucose conditions. AB - Amines such as agmatine, putrescine, spermidine and spermine have been reported to be involved in a variety of physiological and biochemical phenomena. However, it is not known whether they are also involved in the homeostasis of intracellular fibronectin content via upregulation of protein kinase C (PKC), extracellular signal-regulated kinase (ERK), and transforming growth factor-beta1 (TGF-beta1). To determine this, we have studied the effect of multiple amines on fibronectin, TGF-beta1, ERK, and PKC levels in mesangial cells under high glucose conditions. All the amines tested (at 0.1-1 mM) affected neither the viability of mesangial cells for 42 h nor LDH release into the medium. Agmatine reduced TGF beta1 and ERK levels but not PKC at concentrations of 0.1-1 mM. However, levels of fibronectin, TGF-beta1, ERK, and PKC were unaffected by either putrescine or spermidine. A decrease in fibronectin secretion was accompanied by decreases in TGF-beta1 and ERK. Such cumulative results lead us to hypothesize that agmatine reduces high glucose-induced fibronectin secretion via several pathways including ERK-TGF-beta1-fibronectin and spermine, via a decrease in TGF-beta1. Possible roles of enzymes involved in agmatine and polyamine biosynthesis are discussed in relation to secretion of ECM proteins. PMID- 15533579 TI - Short term effects of L-carnitine on serum lipids in STZ-induced diabetic rats. AB - The aim of this study was to evaluate the effects of L-carnitine supplementation on serum triglyceride and total cholesterol levels in streptozotocin (STZ) induced diabetic rats. Thirty-two male Wistar rats were divided into diabetic and diabetic-L-carnitine-supplemented groups. Diabetes was induced by injection of a single dose of streptozotocin (40 mg/kg, intraperitoneally) in citrate buffer. L Carnitine was supplemented by IM injection of 100 mg/kg per day for 10 days. Serum glucose, triglyceride and total cholesterol levels were determined at days 0, 5 and 10. Rats receiving L-carnitine had lower triglyceride levels at both days 5 and 10 (P < 0.05). Total cholesterol levels in the carnitine-supplemented group were lower, but statistical significance was achieved only at day 10 (P < 0.05). These results suggest that L-carnitine exhibits hypotriglyceridemic and hypocholesterolemic effects in streptozotocin-induced diabetic rats. Clinical trials of L-carnitine supplementation on patients with diabetes induced hyperlipidemia must be further evaluated. PMID- 15533580 TI - Short-term effect of buformin, a biguanide, on insulin sensitivity, soluble fraction of tumor necrosis factor receptor and serum lipids in overweight patients with type 2 diabetes mellitus. AB - AIMS/HYPOTHESIS: The UK Prospective Diabetes Study (UKPDS) showed that biguanide therapy in overweight patients reduced the risk for any diabetes-related endpoint and all-cause mortality. Biguanides lower the blood glucose values without stimulation of insulin release. We have investigated the short-term effect of buformin on insulin sensitivity, solved tumor necrosis factor receptors (sTNFRs), and serum lipids in overweight subjects with type 2 diabetes mellitus (DM). METHOD: Thirteen overweight subjects with type 2 DM were examined. The subjects who were fed 20 kcal/kg body weight were divided into two subgroups according to whether they were treated by buformin (Buformin group), or dietary therapy alone (Diet group). Six patients were in Buformin group and seven patients were in Diet group. We calculated insulin-mediated glucose uptake by the liver and peripheral tissues using euglycemic hyperinsulinemic clamp combined with an oral glucose load before and after buformin treatment or diet therapy for 2 weeks. RESULTS: Fasting plasma glucose, total cholesterol (T-chol), LDL-cholesterol (LDL-chol), and sTNFR2 were significantly decreased, and hepatic glucose uptake significantly increased from 32 +/- 7 to 42 +/- 7% (P < 0.05) in Buformin group but did not changed significantly in Diet group. However, the glucose infusion rate thought to express insulin sensitivity in peripheral tissue, TNF-alpha, sTNFR1, fasting plasma insulin, C-peptide, and NEFA levels did not change significantly in both the groups after treatment. CONCLUSION/INTERPRETATION: Buformin improved insulin sensitivity in the liver and decreased T-chol, LDL-chol, and sTNFR2. The mechanism of action for buformin likely involves inhibition of TNF-alpha. Buformin lowers insulin resistance and risk factors for cardiovascular disease including serum lipid and will therefore, be useful in management of overweight type 2 DM patients. PMID- 15533581 TI - Prevalence of gestational diabetes mellitus in Kashmiri women from the Indian subcontinent. AB - This prospective study was carried out to determine the prevalence of gestational diabetes mellitus (GDM) in Kashmiri women and to assess the effect of various demographic factors. Two thousand pregnant women (divided into groups A and B, being the first and last 1000 consecutive women) attending various antenatal clinics in six districts of Kashmir valley were screened for GDM by 1 h 50 g oral glucose challenge test. Four hundred and fourteen (20.8%) women (216 from group A and 198 from group B) had an abnormal screening test and proceeded to oral glucose tolerance testing. Women from group A had a 3 h 100 gram oral glucose tolerance test (OGTT) and GDM was as classified by Carpenter and Coustan. A 2 h 75 g OGTT was performed on group B subjects and WHO criteria applied for diagnosis of GDM. The overall prevalence of GDM was 3.8% (3.1% in group A versus 4.4% in group B-P-value 0.071). GDM prevalence steadily increased with age (from 1.7% in women below 25 years to 18% in women 35 years or older). GDM occurred more frequently in women who were residing in urban areas, had borne three or more children, had history of abortion(s) or GDM during previous pregnancies, had given birth to a macrosomic baby, or had a family history of diabetes mellitus. Women with obesity, hypertension, osmotic symptoms, proteinuria or hydramnios had a higher prevalence of GDM. PMID- 15533582 TI - Do triglycerides explain the U-shaped relation between alcohol and diabetes risk? Results from a cross-sectional survey of alcohol and plasma glucose. AB - The association between alcohol and blood glucose levels, and whether it is modified by other variables, was examined in a cross-sectional survey of 5518 staff aged 40-65 years at worksites in Auckland and Tokoroa, New Zealand. Diabetes was determined by oral glucose-tolerance tests using 1999 WHO criteria. Usual alcohol intake in the previous 3 months, measured by food frequency questionnaire, was related positively with fasting triglycerides and high-density lipoprotein (HDL)-cholesterol, and unrelated with fasting glucose, but had an approximate U-shaped relationship with 2-h glucose, which varied from an adjusted mean (S.E.) of 5.62 (0.08) mmol/l in non-drinkers, down to 5.34 (0.08) mmol/l in light alcohol drinkers (alcohol <5 g per day), and back up to 5.52 (0.09) mmol/l in heavy drinkers (> or =20 g per day). Adjusting further for triglycerides increased the mean difference in 2-h glucose for all drinking categories compared with non-drinkers, particularly for heavy drinkers (> or =20 g per day), from 0.22 (S.E. = 0.10) to -0.37 (0.10) mmol/l. The confounding effect of triglycerides suggests alcohol may affect the diabetes risk by a mechanism related to the triglyceride metabolism, which in heavy drinkers may counteract the protective effect of improved insulin sensitivity, resulting in the U-shaped relationship between alcohol and diabetes described in previous studies. PMID- 15533583 TI - Do diabetologists recognise self-management problems in their patients? AB - OBJECTIVE: The aim of this study was to determine whether diabetologists recognise patients' needs for additional intensive multidisciplinary care due to glycaemic and diabetes-related psychosocial difficulties. RESEARCH DESIGN AND METHODS: We compared 114 participants in a diabetes intervention programme with 201 as yet non-referred outpatients, of whom 54 outpatients were considered eligible for the intervention by their diabetologists; thus, 147 outpatients were considered non-eligible. RESULTS: Analysis revealed that the eligible patients had poorer glycaemic control but all other parameters were similar to non eligible patients. Significantly, 22 (15%) of the 147 non-eligible patients clearly had diabetes distress and could potentially benefit from the intervention. CONCLUSION: The results suggest that in regular care, patients' needs with respect to glycaemic control are recognised by their diabetologists, but patients with high psychosocial diabetes-related distress are often overlooked, though they also may be in need of additional care. Integrated monitoring of diabetes-related distress in outpatients could improve this area of diabetes care. PMID- 15533585 TI - A fourfold difference in the incidence of type 1 diabetes between Sweden and Lithuania but similar prevalence of autoimmunity. AB - We investigated whether other autoimmune disorders in addition to type 1 diabetes are more common in Sweden than Lithuania, and if there are any differences in inheritance patterns of both type 1 diabetes and other autoimmune disorders. Data from 517 children in southeast Sweden and 286 children in Lithuania aged 0-15 years were included in the study. Age- and sex-matched control children were randomly selected. Information was collected by questionnaire. Of the children with diabetes in Sweden, 13.2% had a family member with type 1 diabetes compared to 7% of children with diabetes in Lithuania (P < 0.01) (OR = 2.01). No such difference was seen for other autoimmune diseases in family members of children with diabetes (Sweden 12%, Lithuania 14%, n.s.). Control children in Lithuania had family members with autoimmunity more frequently (15.3%) than control children in Sweden (7.4%, P < 0.001) (OR = 2.26). This difference was most pronounced in mothers. The Lithuanian control children had an autoimmune disease more frequently than the controls in Sweden (4.7% versus 1.5%, respectively, P < 0.001) (OR = 3.21). There seem to be environmental factors that specifically contribute to the development of type 1 diabetes, factors which are less related to the development of autoimmunity in general. PMID- 15533584 TI - Clinical and genetic characteristics of GAD-antibody positive patients initially diagnosed as having type 2 diabetes. AB - The present study was conducted to clarify the clinical and genetic characteristics of the diabetic patients who have antibodies to glutamic acid decarboxylase (GADab) but are diagnosed initially as type 2 diabetes because of the slow progression. Fifty-five GADab+ patients and 137 GADab- patients were recruited. The GADab+ patients were divided into two subgroups according to their antibody titers. The high-titer subgroup (Ab > or = 20 U/ml) had lower urinary C peptide concentrations, and was assigned insulin therapy more often than the GADab- patients. In contrast to the high-titer subgroup, clinical parameters in the low-titer subgroup were similar to the GADab- diabetic patients. The urinary C-peptide levels correlated negatively with the GADab titer in the GADab+ patients. Analysis of type 1 diabetes-susceptible HLA alleles revealed high frequencies of the B54 and DRB1*0405 allele, but not the B61 and DRB1*0901 alleles, in the high-titer subgroup, whereas the frequency of the protective DRB1*1502 allele was decreased. The GADab+ patients with the B54 allele had higher GADab titers and lower urinary C-peptide excretion than patients without this allele. These data indicated that patients with a high-GADab titer share the autoimmune background characteristic of type 1 diabetes. PMID- 15533586 TI - Effects of impaired fasting glucose and impaired glucose tolerance on predicting incident type 2 diabetes in a Chinese population with high post-prandial glucose. AB - AIMS: To ascertain which impaired glucose homeostasis at baseline is predictive of conversion to type 2 diabetes in both men and women in a high-risk non diabetic population. METHODS: The screening survey for type 2 diabetes was conducted in 1994, and a follow-up study of 627 high-risk non-diabetic individuals at baseline was carried out in 1999 in Beijing area. A total of 70 men and 76 women developed diabetes during the 5-year follow-up. RESULTS: After adjustment for age, body mass index, smoking, systolic blood pressure, family history of diabetes, total cholesterol, triglycerides, antihypertensive medication, occupation, education, urine albumin excretion rate, and fasting insulin, the odds ratios for incidence of diabetes were 1.00, 2.04 (95% CI 0.84 4.97), 7.82 (2.66-23.00), 9.38 (3.30-26.69) in men with normal glucose, isolated IFG, isolated IGT, and both IFG and IGT, respectively. In women, the corresponding odds ratios were 1.00, 2.19 (0.91-5.30), 1.62 (0.72-3.67), and 8.79 (2.98-25.93), respectively. CONCLUSIONS: This study shows that combined IFG and IGT at baseline is a strong risk predictor for the development of diabetes in both men and women. Isolated IGT is also a predictor of diabetes in Chinese men. PMID- 15533587 TI - Insulin detemir and insulin aspart: a promising basal-bolus regimen for type 2 diabetes. AB - This trial compared the efficacy and safety of basal-bolus therapy using either the soluble basal insulin analogue insulin detemir (IDet) in combination with meal-time rapid-acting analogue insulin aspart (IAsp), or NPH insulin (NPH) in combination with meal-time regular human insulin (HSI). This was a 22-week, multinational, open-labelled, symmetrically randomised, parallel group trial including 395 people with type 2 diabetes (IDet + IAsp: 195, NPH + HSI: 200). At 22 weeks, HbA1c was comparable between treatments (IDet + IAsp: 7.46%, NPH + HSI: 7.52%, P = 0.515) with decreases from baseline of 0.65% and 0.58%, respectively. Treatment with IDet + IAsp was associated with a significantly lower within person variation in self-measured fasting plasma glucose (FPG) (SD:1.20 versus 1.54 mmol/L, p < 0.001), as well as a lower body weight gain (0.51 versus 1.13 kg, p = 0.038) than with NPH + HSI. The risk of nocturnal hypoglycaemia was 38% lower with IDet + IAsp than with NPH + HSI, but statistical significance was not attained (P = 0.14). The overall safety profile was similar between the two treatments. Basal-bolus treatment with IDet + IAsp is an effective and well tolerated insulin regimen in people with type 2 diabetes, resulting in glycaemic control comparable to that of NPH + HSI, but with the advantages of less weight gain and a lower day-to-day within-person variation in FPG. PMID- 15533588 TI - A case-control study of maternal recreational physical activity and risk of gestational diabetes mellitus. AB - Despite the maternal and infant morbidity associated with gestational diabetes mellitus (GDM), few modifiable risk factors have been identified. We explored the relation between recreational physical activity performed during the year before and during the first 20 weeks of pregnancy and the risk of GDM. 155 GDM cases and 386 normotensive, non-diabetic pregnant controls provided information about the type, intensity, frequency, and duration of physical activity performed during these time periods. Women who participated in any recreational physical activity during the first 20 weeks of pregnancy, as compared with inactive women, experienced a 48% reduction in risk of GDM (odds ratio [OR] = 0.52; 95% confidence interval [CI] 0.33-0.80). The number of hours spent performing recreational activities and the energy expended were related to a decrease in GDM risk. No clear patterns related to distance walked and pace of walking emerged. Daily stair climbing, when compared with no stair climbing, was associated with a 49-78% reduction in GDM risk (P for trend <0.011). Recreational physical activity performed during the year before the index pregnancy was also associated with statistically significant reductions in GDM risk, but women who engaged in physical activity during both time periods experienced the greatest reduction in risk (OR = 0.40; 95% CI 0.23-0.68). These data suggest that recreational physical activity performed before and/or during pregnancy is associated with a reduced risk of GDM. PMID- 15533589 TI - The cell-type-specificity of inherited predispositions to tumours: review and hypothesis. AB - Most hereditary predispositions to tumours affect only one particular cell type of the body but the genes bearing the relevant germ-line mutation are not cell type-specific. Some predisposition syndromes include increased risks of lesions (developmental or tumourous) of unrelated cell types, in any individual predisposed to the main lesion (e.g. osteosarcoma in patients predisposed to retinoblastoma). Other predispositions to additional lesions occur only in members of some families with the predisposition to the basic lesion (e.g. Gardner's syndrome in some families suffering familial adenomatous polyposis). In yet other predisposition syndromes, different mutations of the same gene are associated with markedly differing family-specific clinical syndromes. In particular, identical germline mutations (e.g. in APC, RET and PTEN genes), have been found associated with differing clinical syndromes in different families. This paper reviews previously suggested mechanisms of the cell-type specificity of inherited predispositions to tumour. Models of tumour formation in predisposition syndromes are discussed, especially those involving a germline mutation (the first 'hit') of a tumour suppressor gene (TSG) and a second (somatic) hit on the second allele of the same TSG. A modified model is suggested, such that the second hit is a co-mutation of the second allele of the TSG and a regulator which is specific for growth and/or differentiation of the cell type which is susceptible to the tumour predisposition. In some cases of tumour, the second hit may be large enough to be associated with a cytogenetically-demonstrable abnormality of the part of the chromosome carrying the TSG, but in other cases, the co-mutation may be of 'sub-cytogenetic' size (i.e. 10(2)-10(5) bases). For the latter, mutational mechanisms of frameshift and impaired fidelity of replication of DNA by DNA polyerases may sometimes be involved. Candidate cell-type-specific regulators may include microRNAs and perhaps transcription factors. It is suggested that searching the introns within 10(5)-10(6) bases either side of known of exonic mutations of TSGs associated with inherited tumour predisposition might reveal microRNA cell-type-specific regulators. Additional investigations may involve fluorescent in situ hybridisations on interphase tumour nuclei. PMID- 15533590 TI - Prognostic significance and different properties of survivin splicing variants in gastric cancer. AB - Survivin, a novel antiapoptosis gene, was identified as a member of the inhibitor of apoptosis protein (IAP) family. Unique among IAP, survivin has been found to be abundantly expressed in a wide variety of human malignancies, whereas it is undetectable in normal adult tissues. Recently, three splicing variants of survivin have been further characterized with different subcellular localization, but their different functions in carcinogenesis are largely unknown. We used real time quantitative RT-PCR to analyse survivin variants' mRNA expression levels in 77 gastric carcinoma cases whose frozen samples were available. All the cases and seven cell lines tested expressed wild-type survivin mRNA, which was not only the dominant transcript, but also was a poor prognostic biomarker (P = 0.003). Non antiapoptosis survivin-2B mRNA was negatively correlated with tumor stage (P = 0.001) histological type (P = 0.007) and depth of tumor invasion (P = 0.031) while survivin-DeltaEx3 mRNA showed a significant reverse association with apoptosis ( P = 0.019). These data demonstrated that survivin mRNA expression levels are of important prognostic value, suggesting the significant participation of survivin-2B and survivin-DeltaEx3 in gastric cancer development. PMID- 15533591 TI - Polymorphisms of XRCC1 and risk of esophageal and gastric cardia cancer. AB - BACKGROUND: Linxian, a rural county in North Central China, has among the highest rates of esophageal squamous cell carcinoma and gastric cardia adenocarcinoma in the world. In a nested case-cohort study that originated from two cancer prevention trials in Linxian, we examined the relationship between these cancers and two polymorphisms in the DNA repair gene XRCC1. METHODS: We conducted a case cohort study among individuals in the cohort who were alive and cancer free in 1991, and had blood samples for DNA extraction. Real time Taqman analyses were conducted to genotype incident cancer cases (n = 221, 131 esophageal and 90 gastric cardia cancer cases) that developed through May 1996, and on an age- and sex-matched reference cohort (n = 454). We used Cox proportional hazard models to estimate relative risks (RR) and 95% confidence intervals (95% CI). RESULTS: We observed no association between the variant genotype in XRCC1 Arg194Trp (codon 194 arganine to tryptophan substitution) and esophageal or gastric cardia cancer. However, carrying at least one copy of the variant allele in XRCC1 Arg399Gln (codon 399 arganine to glutamine substitution) was associated with reduced risk of gastric cardia cancer (RR: 0.60, 95% CI: 0.37-0.97) and the combined category esophageal/gastric cancer (RR: 0.67, 95% CI: 0.48-0.95). In combined polymorphisms analyses, we observed a significant reduction in risk of combined esophageal/gastric cancer among individuals that had both the XRCC1 Arg194Trp and Arg399Gln variant genotyopes (RR: 0.47, 95% CI: 0.26-0.84). CONCLUSIONS: Our results suggest that the XRCC1 Arg399Gln variant genotype is associated with reduced risk of upper GI cancer and that individuals with both XRCC1 variant genotypes are also at significantly reduced risk of upper GI cancer in this high risk Chinese population. PMID- 15533592 TI - Beauvericin induces cytotoxic effects in human acute lymphoblastic leukemia cells through cytochrome c release, caspase 3 activation: the causative role of calcium. AB - Beauvericin (BEA), a cyclic hexadepsipeptide, induces cell death in human leukemia cells (CCRF-CEM) and the process of BEA-induced cell death has been speculated to undergo an apoptotic pathway. In the present study, several well characterized factors, known to play important roles in apoptotic pathway, were investigated in BEA-induced CCRF-CEM cell death. CCRF-CEM cells were treated with BEA at concentrations from 1 to 10 microM for up to 24 h. The incidence of nuclear fragmentation and apoptotic body formation in the cells, cytosolic caspase-3 activity, mitochondrial membrane potential, and release of cytochrome c (Cyt c) from mitochondria in BEA-treated cells were determined and compared with that in untreated cells. Moreover, to investigate the role of intracellular Ca++ in this cell death process, CCRF-CEM cells were primed with 3 microM of BAPTA/AM, a Ca++ chelator, to exclude intracellular Ca++ prior to the BEA treatment. The data revealed that BEA-induced cell death in CCRF-CEM cells exhibited a dose- and time-dependent manner. The incidence of nuclear fragmentation and apoptotic body formation was significantly increased in CCRF-CEM cells treated with BEA at concentrations of 1 microM or greater. Increase of cytosolic caspase-3 activity was also observed in BEA-treated cells with a dose-dependent manner. In addition, increased release of Cyt c from mitochondria was also observed in the cells treated with 10 microM BEA in a time-dependent pattern. The BAPTA/AM pretreatment partially blocked BEA-induced cell death in CCRF-CEM cells, indicating that intracellular Ca++ plays an important role, maybe as a mediator in cell death signaling, in this cell death pathway. The results support the notion that BEA induced cell death in CCRF-CEM cells likely undergo through an apoptotic pathway on the basis of increase of release of Cyt c from mitochondria, increase of caspase-3 activity, and some observed typical apoptotic cellular changes in morphology. PMID- 15533593 TI - Protein-bound polysaccharide from Phellinus linteus induces G2/M phase arrest and apoptosis in SW480 human colon cancer cells. AB - The cytotoxic mechanism of protein-bound polysaccharide isolated from Phellinus linteus (PL, Mesima) has been investigated. PL inhibited the proliferation and colony formation of SW480 human colon cancer cells. Flow cytometry analysis showed that PL increased the populations of both apoptotic sub-G1 and G2/M phase. The result obtained from TUNEL assay corroborated apoptosis which was shown in flow cytometry. Western blot analysis suggested that PL-induced apoptosis and growth inhibition were associated with decrease in Bcl-2, increase of the release of cytochrome c, and reduced expression of cyclin B1. These results suggest that PL has a direct antitumor effect through apoptosis and cell cycle blockade in certain cancer cells. PMID- 15533594 TI - Calpains are activated by photodynamic therapy but do not contribute to apoptotic tumor cell death. AB - Photodynamic therapy (PDT) of cancer is a promising technique based on the formation of singlet oxygen following irradiation of a sensitizer with visible light. In the present work we investigated the role of calpains in PDT, using the human lymphoblastoid CCRF-CEM cells and bisulfonated aluminum phthalocyanine (AlPcS2) as a sensitizer. Photosensitization induced apoptotic cell death and a time-dependent activation of calpains, as determined using the fluorogenic substrate succinyl-Leu-Leu-Val-Tyr-7-amido-4-methylcoumarin (SLLVY-AMC). However, inhibition of calpains with calpain inhibitor II or with PD 150606 did not affect the demise process. The results indicate that although calpains are activated in PDT, they do not play a major role in tumor cell death. PMID- 15533595 TI - Associations between UVR exposure and basal cell carcinoma site and histology. AB - While sunlight is critical in basal cell carcinoma (BCC) pathogenesis the relationship between exposure and tumor site and histology is unclear. We determined if tumor site (trunk or head/neck) or histology (nodular or superficial) is determined by exposure pattern. In 66 cases with truncal and 362 patients with head/neck BCC at first presentation, average hours exposure/year, intermittency score, childhood sunburning and skin type were not significantly associated with tumor site or histology. However, often sunbathing was associated with a five-fold increased risk of truncal BCC. Average sunbathing score was significantly greater in 22 cases with truncal compared with 325 cases with head/neck nodular tumors and also in 44 cases with superficial truncal compared with superficial head/neck BCC. Thus, sunbathing determined tumor site but not histology. PMID- 15533596 TI - Methylenetetrahydrofolate reductase polymorphism, alcohol intake, and risks of colon and rectal cancers in Korea. AB - Several, but not all, studies have reported that a variant genotype of the polymorphism (C677T) of 5,10-methylenetetrahydrofolate reductase (MTHFR), an enzyme in folate metabolism, is associated with a decreased risk of colorectal cancer. A case-control study was conducted to investigate the association of MTHFR polymorphism and heavy alcohol intake to colon and rectal cancer in Korean. Cases were a consecutive series of patients with histologically confirmed, incident colorectal cancer who were admitted to two university hospitals in Seoul, Korea between 1998 and 2000, and controls were selected at the same hospitals. A total of 243 cases (colon 111, rectum 132) and 225 controls were enrolled. While the genotype of MTHFR was not associated with the overall risk of colorectal cancer, increased colon cancer risk was found to be associated with the CT and TT genotypes combined (multivariate odds ratio [OR] 2.01, 95% confidence interval [CI] 1.14-3.53) compared with the wild type. The risk of rectal cancer was found to be, though statistically non-significant, lower in those with the CT and TT genotypes combined (multivariate OR 0.67, 95% CI: 0.43 1.07). Those consuming two or more drinks per day (30 g+/day) had nearly twice the colorectal cancer risk (multivariate OR 1.94, 95% CI 1.03-3.68) of light or non-drinkers (<5 g/day). The present study did not find a reduced risk of colorectal or rectal cancer among those with a variant genotype of the MTHFR polymorphism, but observed rather an increased risk of colon cancer, suggesting that the effects of the MTHFR genotype may differ in populations with different levels of folate intake. PMID- 15533597 TI - Expression of glutathione S-transferase P1-1 in leukemic cells is regulated by inducible AP-1 binding. AB - Glutathione S-transferases (GST) are involved in cellular protection against xenobiotics, oxidative stress as well as in resistance against chemotherapeutic compounds such as doxorubicin. Levels of human placental type GSTP1-1 are known to be increased in many tumors and hematopoietic diseases. In this work, we compare transcriptional mechanisms in cells that express or not GSTP1-1. Transient transfection assays are used to show that different GST-promoter reporter constructs generate cell-type specific levels of luciferase activity. In expressing cells, transcriptional activity is strongly dependent on AP-1 binding elements within the -65 to -75 bp region of the GSTP1 gene as shown by site directed mutagenesis. Electrophoretic mobility shift assays show that DNA binding activity is exclusively observed in GSTP1-1-expressing cells and is increased after stimulation with hydrogen peroxide, TPA, tert-butylhydroquinone and doxorubicin. Non-expressing cells present neither constitutive nor inducible AP-1 binding. Taken together, our results provide evidence for the induction of the GSTP1 gene via AP-1 binding activity in leukemia cells and contribute to a better understanding of the molecular events regulating genes involved in drug resistance mechanisms. PMID- 15533598 TI - Topoisomerase II gene mutations in tumors and tumor cell lines with microsatellite instability. AB - Genetic or epigenetic inactivation of the DNA mismatch repair genes in tumor precursor cells results in a strong mutator phenotype, known as the microsatellite mutator phenotype (MMP), or microsatellite instability (MSI). This mutator phenotype causes mutations in genes responsible for the regulation of cell growth and survival/death and thus promotes the development and progression of tumors. In the present study, we examined the DNA topoisomerase II genes (topIIalpha and topIIbeta) as mutational targets for MMP. We screened 10 MSI positive human tumor cell lines and 30 MSI-positive colorectal tumors for mutations within the entire coding region of the topIIalpha gene and two coding poly(A)7 sequences of topIIbeta. Mutations in either the topIIalpha or topIIbeta gene were found with an overall frequency of 18% (in 10% of the primary tumors and in 44% of the cell lines). This indicates that modulation of the DNA topoisomerase II (TOPII) activity may be important for the development of MSI positive cancer. PMID- 15533599 TI - The cancer recognition (CARE) antibody test. AB - The cancer recognition (CARE) antibody (Ab) test is a serologic assay for a specific IgM that is elevated in cancer patients. All tests are measured using an indirect enzyme-linked immunosorbent assay (ELISA) of human serum. The target polypeptide in the CARE Ab test is the IgM binding epitope (LT-11) of the CARE antigen (Ag) consisting of a 16 mer structure that has been produced synthetically. The mean relative concentration (MRC) is determined relative to standard, normalized human plasma. Non-parametric analysis showed median MRC values of healthy volunteers (HVs) with no history of cancer (n =47), family history of cancer (n = 126) and a previous cancer history (n = 24) to be 26, 34 and 46, respectively. It was determined that there was no significance found among the medians of the three HV groups (P = 0.53). The specificity of the HV types was between 87 and 98%. Benign/non-cancer surgical patients (n = 27) had a median value of 20 with a specificity of 96%. The cancer patients (n = 61) had a median value of 246 with a sensitivity of 89%. There was a significant difference between the HV and cancer patients (P < 0.0001) as well as between the benign/surgical non-cancerous group and cancer patients (P < 0.0001). The IgM antibody is heat stable at room temperature for two days versus being frozen at 80 degrees C (r2 = 0.97). Either serum or plasma samples may be used in the CARE Ab test (r2 = 0.92). The CARE Ab was almost exclusively IgM with no serum conversion to IgG in sequential measurements of patients with cancer over a six month period. Preliminary data from patients undergoing post-operative cancer treatment showed that decreasing Ab levels revealed patients negative for residual cancer or undergoing remission, while relapsing patients show an increase in Ab levels. A return to a positive Ab level shortly after treatment is a poor prognostic sign while in advanced cancers the Ab levels may be depressed significantly. PMID- 15533600 TI - Metabolism of the microregions of human breast cancer. AB - Glucose and glutamine metabolism of two microregions of human infiltrating ductile breast cancer, the center and the periphery, was studied and the results were compared with those of healthy mammary glands. In general, the activities of glycolytic enzymes and of phosphate-dependent glutaminase were as follows: center>periphery>mammary gland. Insulin caused a marked increase of glucose consumption and lactate production by incubated slices of mammary gland but had no effect on both microregions of the tumor. Therefore, human breast cancer presents metabolic microregions. PMID- 15533601 TI - Suppression of the acute-phase response as a biological mechanism for the placebo effect. AB - The idea that inert substances such as sugar pills can have powerful therapeutic effects--the so-called 'placebo effect'--has been widely accepted by most medical researchers since the 1950s. Today there is increasing scepticism about the reality of the placebo effect. This debate has been too simplistic; rather than asking whether or not the placebo effect exists, as researchers have done up to now, we should be more precise, and ask which medical conditions (if any) are placebo-responsive. There is evidence that pain, swelling, stomach ulcers, depression, and anxiety are all placebo-responsive. These conditions have all been linked, to a greater or lesser extent, with activation of the acute-phase response (the innate immune response). The placebo effect may therefore be mediated by alteration of one or more components of the acute-phase response. The candidates for such biochemical mediators would need to alter the synthesis, activation, receptor-binding or signalling mechanisms of inflammation, sickness behaviour and other aspects of innate immunity. This hypothesis is consistent with current data suggesting that placebos work by triggering the release of endorphins. The hypothesis would be falsified if it were found that other medical conditions, not involving the activation of the acute-phase response, were nonetheless alleviated by placebos. PMID- 15533602 TI - Paroxysmal cough injury, vascular rupture and 'shaken baby syndrome'. AB - It is widely assumed that subdural and retinal haemorrhage in infants can only result from traumatic rupture of vulnerable blood vessels. An alternative aetiology, that of vascular rupture resulting from excessive intraluminal pressure, is presented in three disease conditions. (1) Perlman et al., studying premature neonates requiring mechanical ventilation for respiratory distress syndrome, observed "cough-like" fluctuations in oesophageal pressure greater than 18 cms H2O, whose timing matched fluctuations in anterior cerebral artery flow. When 14 out of 24 neonates were paralysed (to prevent abdominal muscle activity) intraventricular haemorrhage developed in all 10 controls but in only one of the paralysed group during paralysis. (2) New analysis of pressure data extracted from a previous study of prolonged expiratory apnoea showed alveolar collapse induced 100 mmHg intrathoracic cough pressure surges. Superior vena cava pressures up to 50 mmHg were implied, and radial artery systolic pressures over 180 mmHg recorded. (3) Bordetella pertussis bacteria attach to cilia in the airways, but do not invade the underlying tissue. The irritation causes the powerful coughing paroxysms of whooping cough. Brain haemorrhages and retinal detachment have been observed to result from the high intravascular pressures produced. The data suggest that any source of intense airway irritation not easily removed (laryngeal infection, inhalation of regurgitated feed, fluff, smoke etc.) could induce similar bleeding, a paroxysmal cough injury (PCI). Additional objective evidence of inflicted trauma is necessary to distinguish between 'shaken baby syndrome' and PCI. PMID- 15533603 TI - Antibiotics may act as growth/obesity promoters in humans as an inadvertent result of antibiotic pollution? AB - The growth promoting effects of antibiotics were first discovered in the 1940s. Since then, many antimicrobials have been found to improve average daily weight gain and feed efficiency. The total production of antibiotics can be estimated between 100,000-200,000 tons annually and the human population is being influenced, directly or indirectly (from the environment) by this amount of drug. The twentieth-century increase in human height and the obesity of the population is roughly observed since the mass consumption of antibiotics 40-50 years ago. The association between antibiotic consumption and the increase of human growth/obesity is suspected. PMID- 15533604 TI - Direct placental effects of cigarette smoke protect women from pre-eclampsia: the specific roles of carbon monoxide and antioxidant systems in the placenta. AB - Pre-eclampsia is a hypertensive disorder of pregnancy characterized by shallow placentation, inadequate placental perfusion, localized placental oxidative stress, a heightened maternal inflammatory response and subsequent maternal endothelial dysfunction. This pathophysiology leads to an increase in maternal blood pressure, edema and proteinurea. Interestingly, women who smoke cigarettes throughout pregnancy are at a 33% reduced risk of developing this disorder. The exact mechanisms through which cigarette smoke reduces the risk of pre-eclampsia are not yet understood. We propose that cigarette smoke reduces the risk of developing pre-eclampsia via direct placental effects. In this review we will address, and provide evidence for, our specific hypotheses that: (a) CO increases trophoblast invasion and spiral arteriole remodeling; (b) CO decreases a localized inflammatory response at the level of the decidua; (c) CO increases utero-placental, intra-placental and feto-placental blood flow; (d) CO decreases hypoxia-induced apoptosis of the syncitiotrophoblast layer; (e) CO activates hemoproteins involved in normal endothelial functioning normally acted upon by NO; (f) compound(s) within cigarette smoke result in upregulation of antioxidant systems within the placenta. These various mechanisms of action must be further examined as they may provide valuable keys to novel therapeutic design in the realm of pre-eclampsia research. PMID- 15533605 TI - Serotonergic hypothesis of sleepwalking. AB - Despite widespread prevalence of sleepwalking, its etiology and pathophysiology are not well understood. However, there is some evidence that sleepwalking can be precipitated by sleep-disordered breathing. A hypothesis is proposed that serotonergic system may be a link between sleep-disordered breathing and sleepwalking. Serotonergic neurons meet basic requirements for such a role because they are activated by hypercapnia, provide a tonic excitatory drive that gates afferent inputs to motoneurons, and the activity of serotonergic neurons can be dissociated from the level of arousal. This paper discusses also drug induced somnambulism and co-occurrence of sleepwalking and other disorders such as migraine and febrile illness. PMID- 15533606 TI - Can SSRIs reduce the risk of preeclampsia in pregnant, depressed patients? AB - Preeclampsia causes substantial morbidity and mortality. A significant part of the etiology of preeclampsia involves endothelial damage and platelet activation and in this way can be conceived as an illness with a pathophysiology similar to coronary artery disease. Depression is an independent risk factor for the progression of cardiovascular disease [Am Heart J 140 (2000) S57] and there is evidence to suggest that it may be a risk factor for preeclampsia as well. SSRIs have been shown to reduce the progression of coronary artery disease, independent of improvement in mood. SSRIs may also reduce risk factors for preeclampsia in addition to treating depression in pregnancy. This is an important area for further research. PMID- 15533607 TI - Could inguinal hernia be treated medically? AB - Inguinal hernia is the most common congenital anomaly requiring surgical correction. The cause of the hernia is, most commonly, persistence of the processus vaginalis. Study of testicular descent in rodents has revealed a role for the genitofemoral nerve and calcitonin gene-related peptide (CGRP). Since the testis cannot descend without the processus vaginalis, we wondered whether both descent and hernia closure might be regulated by the same mechanism. Therefore, we tested the idea that CGRP might be active in closure of the inguinal hernia. Using hernial sacs removed at herniotomy operation, fusion of the hernial surfaces was induced by CGRP, but not by CGRP 8-37, CGRP 27-37 or dihydrotestosterone. Hepatocyte growth factor also caused peritoneal obliteration in vitro. We propose that obliteration of the processus vaginalis is under the control of molecules released from the genitofemoral nerve, and that a chemical treatment of inguinal hernia is at least theoretically possible. PMID- 15533609 TI - The putative role of apoptosis in the induction of pemphigus. AB - The apoptotic cell has recently been shown to play a central role in tolerizing B cells and T cells to both tissue-specific and ubiquitously expressed self antigens, and to possibly drive the autoimmune response in systemic lupus erythematosus, an autoimmune disease which bears many similarities to pemphigus. We now propose a similar mechanism in the induction of pemphigus, namely, that a dysregulation in apoptosis expressed as an impairment of normal programmed cell death of epidermal keratinocytes and/or deficient and inadequate clearance of apoptotic material (specifically, desmoglein) may render it antigenic with the consequent production of autoantibodies. The fact that some thiol-containing compounds which are well-known inducers of acantholysis in vitro and pemphigus in vivo were shown to inhibit apoptosis might support our hypothesis. PMID- 15533608 TI - Exacerbation of rheumatoid arthritis following Helicobacter pylori eradication: disruption of established oral tolerance against heat shock protein? AB - A 62-year-old Japanese woman with RA received an eradication therapy against Helicobacter pylori in November 1999. Eight weeks later, successful eradication was confirmed by negative results for rapid urease test, pathologic findings, and a fall in anti-H. pylori IgG antibody titer. During the course, parameters for RA activity were exacerbated: C-reactive protein 1.1-4.2 mg/dL, rheumatoid arthritis precipitation antigen 2560-5120 dils., erythrocyte sedimentation rate 52-123 mm/h, and complements CH50 50 to over 60 U/mL. Lansbury index increased from 70% to 105%. Two more weeks later, the patient noticed right shoulder pain. She also complained of bilateral gonalgia two months later, and physical examination revealed increased fluid in the knee joints. Prednisolone was required to control the disease activity. The results of this case suggested that RA patients might experience a deleterious effect on the disease activity following H. pylori eradication possibly through disruption of the established oral tolerance against stress protein such as mycobacterial heat shock protein 65. PMID- 15533610 TI - Iloprost: an adjunctive approach to chronic viral hepatitis treatment. AB - Chronic viral liver disease may evolve to cirrhosis. The medical treatment to slow down this passage is based on anti-viral and anti-fibrotic properties of interferon. Recently, we evidenced significant increase of portal vein flow velocity and volume after a prostacyclin analog (iloprost) infusion in subjects without and with chronic viral hepatitis. On the basis of these results and considering both the pathophysiology of viral liver disease and the mechanism of action of iloprost in portal microcirculation, we hypothesize that it may be of some efficacy in chronic liver disease ameliorating the portal hemodynamics and producing an anti-oxidant liver effect. PMID- 15533611 TI - Statins lower the risk of developing Alzheimer's disease by limiting lipid raft endocytosis and decreasing the neuronal spread of Herpes simplex virus type 1. AB - Many possible risk factors for Alzheimer's disease (AD) have been investigated, with only a very few showing positive associations and none defining the etiology of the neurodegenerative disease. The presence of herpes simplex virus type 1 (HSV-1) DNA in the brain, coupled with apolipoprotein E allele e4 (ApoE e4), has been suggested to confer an increased risk for AD. Studies have shown that pathogens, including viruses, utilize clathrin-independent endocytosis, i.e., lipid rafts that contain cholesterol, as part of their structure. Moreover, cholesterol-lowering statins have recently been linked with a reduced risk of developing Alzheimer's dementia. We, therefore, posit that long-term statin therapy protects individuals from AD by reducing the neuronal spread of HSV-1 via lipid raft domain pathways. Although the mechanism by which statins reduce AD risk is unknown, they reduce the amount of cholesterol in the plasma membrane and, thus, may decrease the availability of lipid raft pathways to spread HSV-1 within the brain. PMID- 15533612 TI - Is there a role for long-chain omega3 or oil-rich fish in the treatment of atrial fibrillation? AB - Atrial fibrillation is the most common cardiac arrhythmia in Europe and north America, and recently it was described as an epidemic. Treatment and management of this arrhythmia consists of using drugs, external electrical cardioversion and in extreme cases, internal electrical pacing. Despite treatment, this arrhythmia continues to impact on morbidity and mortality. The possible benefit from dietary interventions in relation to the primary and secondary prevention of atrial fibrillation have largely been overlooked. Our hypothesis is that increasing the intake of long-chain polyunsaturated omega3 fatty acids (LCn3) from eating a diet containing moderate amounts of oil-rich fish, will benefit people with persistent atrial fibrillation. A number of possible anti-arrhythmic actions from LCn3 have been found from animal and laboratory studies, mainly on ventricular arrhythmias. These include reducing pro-arrhythmic eicosanoids and inhibiting sodium and calcium currents. If found to be beneficial to these patients, dietary advice to eat more oil-rich fish, or take LCn3 supplements, could be part of a package of care for people with this arrhythmia. We have currently started a randomised controlled trial to test our hypothesis. PMID- 15533613 TI - Effect of antibiotics, prebiotics and probiotics in treatment for hepatic encephalopathy. AB - In order to reduce ammonia production by urease-positive bacteria Solga recently hypothesised (S.F. Solga, Probiotics can treat hepatic encephalopathy, Medical Hypotheses 2003; 61: 307-13), that probiotics are new therapeutics for hepatic encephalopathy (HE), and that they may replace antibiotics and lactulose. This influenced our view of the effect of antibiotics, prebiotics, e.g., lactulose, and probiotics on intestinal bacteria in the treatment of HE. Intestinal ammonia arises from aminoacids after bacterial de-amination and not from urea making urease-positive bacteria irrelevant. Antibiotics are not preferred in the treatment of HE, since ammonia-producing antibiotic-resistant bacteria may survive and replace ammonia-producing antibiotic-susceptible bacteria. Intestinal prebiotics are carbohydrate-like compounds, such as lactulose and resistant starch, that beneficially affects host's health in a different manner than normal food. In the small bowel prebiotics are not absorbed and digested, but are fermented in the colon by colonic bacteria. Fermentation of prebiotics yields lactic, acetic and butyric acids, as well as gas especially hydrogen (H2). The massive H2 volumes cause rapid intestinal hurry and thus massive amounts of colonic bacteria, not only urease-positive bacteria, but also deaminating bacteria, are removed and intestinal uptake of toxic bacterial metabolites, e.g., ammonia, reduced. As living non-pathogenic micro-organisms, probiotics beneficially affect the host's health by fermenting non-absorbed sugars, especially in the small bowel. Thus, they reduce the substrate of the other bacteria, and simultaneously they create a surplus of fermentation products which may affect the non-probiotic flora. Regarding the fermentation products (lactic acid, ethanol, acetic acid and CO2) five groups of probiotic micro-organisms are known. It is argued that probiotic, CO2-producing (facultatively) heterolactic lactobacilli, i.e., lactobacilli, that produce both lactic acid and CO2 from sugars, such as glucose, are preferred in the treatment of HE. Our ideas concur with the practice guidelines regarding HE as formulated by Blei, Cordoba and the Practice Parameters Committee of the American College of Gastroenterology, and does not alter the final conclusion of Solga as regards the beneficial use in future treatment of HE. PMID- 15533614 TI - The role of fat tissue in the cholesterol lowering and the pleiotropic effects of statins--statins activate the generation of metabolically more capable adipocytes. AB - Statins have very important influences on adipocyte physiology, particularly via their effects on sterol regulatory element binding proteins (SREBPs), which are extremely important transcription factors for the regulation of adipocyte physiology. SREBP-1c activation in fat cell by the cytosolic cholesterol deprivation through HMG-reductase inhibition is responsible for most of the beneficial effects of statins. Glucose and lipid excess, and oxidative overload coming with energy dense nutrition, which might be deleterious, can be removed from the circulation by adipo-lipogenetic enhancement through the activated SREBP 1c. SREBP-1c causes formation of new small and competent adipocytes, which are insulin and leptin sensitive. Thus, adipocytes gain the ability of fat storing, and much more importantly, fat-burning machines. Statins already have a large and growing number of indications in the treatment of atherovascular diseases. General mechanism for the action of statins is their inhibitory effects on 3 hydroxy 3-methyl glutaryl CoA reductase, which is the rate-limiting enzyme in cholesterol biosynthesis, particularly in the liver. Recently, the beneficial effects of statins beyond their cholesterol lowering, which are called pleiotropic effects, have been the focus of great attention. Certain characteristics of the statin molecule itself have been proposed to explain these effects. Adipocytes are in the very heart of metabolic regulations. Therefore, the important elements of the pleiotropic effects of statins occur through executive improvements on impaired adipocyte functions by the activation of SREBP 1c. PMID- 15533615 TI - How PEDF prevents angiogenesis: a hypothesized pathway. AB - Pigment epithelium-derived factor (PEDF) is a multiple functional protein, coded by the serine proteinase inhibitor, clade F, member 1 (SERPINF1) gene, which has both anti-angiogenic activity and neurotrophic activity at the same time. Its antiangiogenic activity in the mammalian eye is the most potent known at this time. However, the mechanism(s) by which PEDF works in vivo is still uncertain. Some observations suggest that PEDF can simultaneously inhibit the migration and proliferation induced by vascular endothelial growth factor (VEGF), and then further inhibits angiogenesis by interacting with specific cell surface receptors, but no such receptor has been reported to date. Here we propose a hypothesis that PEDF exerts its function by binding with intergrins. Intergrin can therefore serve as the receptor of PEDF. PMID- 15533616 TI - Bariatric barosensor: a new paired structure in the paravetebral region of the iliac crest. AB - A new anatomical entity, which we have decided to name bariatric borosensor, is described. It represents a paired structure under the skin, supero-lateral to the posterior superior iliac spine, overlying the iliac crest at the level of the fourth lumbar vertebra. Each of the paired structure is about the size of an enlarged lymph node; 0.5-2.0 cm in the largest diameter, spherical or ovoid in shape and firm in consistency. They are easily palpated clinically and can be demonstrated objectively by ultrasound examination. A hypothesis is put forth, conjecturing that this new structure may have relevance to the problem of obesity in modern, civilised western societies. PMID- 15533617 TI - Functional similarities between pleura and the renal proximal tubule--membrane and cellular considerations. AB - The small amount of liquid that, under physiological conditions, is presented in the pleural cavity has been the focus of extensive research for more than a century. However, there are still unanswered questions and considerable controversies about the nature of the forces governing its movement into and out of the pleural cavity. Early in the 20th century has been proposed that pleural fluid turnover is simple based on the balance between hydraulic and colloid osmotic pressures existing across the pleural membranes. This original hypothesis has not been validated by data accumulating over the last 20 years. Pleural tissues and renal proximal tubules present high water permeability, small transepithelial electrical resistance (22.02 Omega cm2) and the same cation transportation such as Na+ channels, Na+-K+ ATPase channels, and Na+-H+ exchanger. In contrast to previous conflicting theories concerning pleura fluid movement, the same functional characteristics suggest the hypothesis that physiology of pleura is similar to proximal tubules. PMID- 15533618 TI - Relationship between retinal lattice degeneration and open angle glaucoma. AB - Patients with retinal disorders may develop glaucoma of both a primary and secondary type. Pigment may contribute to trabecular obstruction in some patients with open-angle glaucoma. Lattice degeneration of the retina in its typical form is a sharply demarcated, circumferentially oriented, degenerative process with significant alterations of retinal pigmentation. The association between myopia, open angle glaucoma and pigment dispersion is striking. Therefore, it could be postulated that there is significant prevalence of open angle glaucoma in patients with retinal lattice degeneration, especially in combination with myopia. PMID- 15533619 TI - 3-D physical models of amitosis (cytokinesis). AB - Based on Newton's laws, extended Coulomb's law and published biological data, we develop our 3-D physical models of natural and normal amitosis (cytokinesis), for prokaryotes (bacterial cells) in M phase. We propose following hypotheses: Chromosome rings exclusion: No normally and naturally replicated chromosome rings (RCR) can occupy the same prokaryote, a bacterial cell. The RCR produce spontaneous and strong electromagnetic fields (EMF), that can be alternated environmentally, in protoplasm and cortex. The EMF is approximately a repulsive quasi-static electric (slowly variant and mostly electric) field (EF). The EF forces between the RCR are strong enough, and orderly accumulate contractile proteins that divide the procaryotes in the cell cortex of division plane or directly split the cell compartment envelope longitudinally. The radial component of the EF forces could also make furrows or cleavages of procaryotes. The EF distribution controls the protoplasm partition and completes the amitosis (cytokinesis). After the cytokinesis, the spontaneous and strong EF disappear because the net charge accumulation becomes weak, in the protoplasm. The exclusion is because the two sets of informative objects (RCR) have identical DNA codes information and they are electro magnetically identical, therefore they repulse from each other. We also compare divisions among eukaryotes, prokaryotes, mitochondria and chloroplasts and propose our hypothesis: The principles of our models are applied to divisions of mitochondria and chloroplasts of eucaryotes too because these division mechanisms are closer than others in a view of physics. Though we develop our model using 1 division plane (i.e., 1 cell is divided into 2 cells) as an example, the principle of our model is applied to the cases with multiple division planes (i.e., 1 cell is divided into multiple cells) too. PMID- 15533620 TI - Pro PSA : a "pro cancer" form of PSA? AB - Pro PSA, the precursor of PSA, is an inactive 244-amino acid protein secreted by prostatic cells. Pro PSA, a distinct molecular form of free PSA in serum, includes native as well as several truncated forms. Clinical studies have recently provided evidence that pro PSA is associated with prostate cancer (PCa). The truncated (-2) pro PSA form accounts for only 6-19% of free PSA in the serum of patients without PCa, but it represents up to 25-95% of free PSA in that of PCa patients. In the PSA range of 2-10 ng/ml, it has been suggested that pro PSA may be useful to detect PCa, and to spare 10-20% of unnecessary prostate biopsies. However, only scant information about the role of pro PSA is available to date, and its real impact on PCa detection remains to be determined. Furthermore, it is questionnable whether pro PSA could be predictive of tumor recurrence of PCa after therapy. Prospective clinical studies with long term followup are needed to clarify this point. PMID- 15533621 TI - Maternal heterozygosity for a mitochondrial trifunctional protein mutation as a cause for liver disease in pregnancy. AB - Acute fatty liver of pregnancy (AFLP) and hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome are serious complications of pregnancy associated with significant maternal and perinatal morbidity and mortality. In previous reports, we have documented an association between AFLP and fetal deficiency of long-chain 3-hydroxyacyl coenzyme A dehydrogenase (LCHAD) [N. Engl. J. Med. 340 (1999) 1723-1731; JAMA 288 (2002) 2163-2166]. LCHAD activity resides in the alpha subunit of the mitochondrial trifunctional protein (MTP), a complex protein that catalyzes beta-oxidation of long chain fatty acids. In all reported cases, the fetus carried a common alpha-subunit MTP mutation (G1528C, E474Q) on one or both alleles. However, the association between fetal LCHAD deficiency and the maternal HELLP syndrome has been limited. Here, we report a case history of a 27-year-old black female who underwent Cesarean section for placenta previa and fetal distress at 36 weeks gestation. The newborn was a healthy male child. Post delivery, the mother developed severe HELLP syndrome with complications resulting in death of the patient. We used single strand conformation variance and nucleotide sequence analyses to screen DNA isolated from the mother and the newborn for mutations in the MTP alpha-subunit. The mother was heterozygous for a novel mutation (C1072A, Q322K) in exon 11 of the LCHAD domain of the MTP, while the fetal genotype was completely normal. We hypothesize that, in some cases, maternal heterozygosity for an MTP mutation maybe sufficient to cause the development of maternal liver disease without carrying an affected fetus. Combination of the metabolic stress of pregnancy and other environmental stresses may overwhelm the heterozygous mother's capacity for effective metabolism of long chain fatty acids, leading to an accumulation of potentially toxic fatty acid metabolites in the maternal circulation with subsequent damage to the maternal liver. PMID- 15533622 TI - Role of high-insulinogenic nutrition in the etiology of gestational diabetes mellitus. AB - A transient physiologic insulin resistance and hyperinsulinemia are characteristic of normal pregnancy. This insulin action has evolved during a period of human evolution that was characterized by a very low-carbohydrate nutrition. The development of gestational diabetes mellitus (GDM) is proposed to result from a collision of this evolutionary inheritance with our "modern" nutrition: The "Western" high-insulinogenic nutrition increases the postprandial demand for insulin significantly during the insulin resistant state of late pregnancy. In women with beta-cells that are not capable of maintaining the high insulin production, GDM develops. A restriction of high-insulinogenic carbohydrate may help to prevent the development of GDM. PMID- 15533623 TI - Iminium metabolite mechanism for nicotine toxicity and addiction: Oxidative stress and electron transfer. AB - The mechanism of nicotine toxicity is not completely delineated. Considerable evidence points to involvement of oxidative stress (OS), reactive oxygen species, lipid peroxidation, DNA damage, and beneficial effect of antioxidants. Previously, a suggestion was advanced for participation of iminium metabolites which might operate, via electron transfer (ET) with redox cycling, to produce radical entities. The conjugated iminium functionality is one of the less well known ET types. The cationic metabolites arise from several routes, including oxidation of nicotine itself, and protonation of myosmine which originates from nornicotine via demethylation of nicotine. Reduction potentials, which are in the range amenable to ET in vivo, lend credence to the theoretical framework. An alternate metabolic route entails hydrolysis of nicotine iminium to an open-chain ketoamine that, in turn, undergoes nitrosation to form a toxic nitrosamine. Subsequently, the nitrosamine serves as a DNA alkylator which can also generate conjugated iminiums by attack on certain nitrogens of DNA bases. During the past 14 years, the hypothesis has enjoyed substantial support. Increasing evidence points to a role for OS in toxicity by nicotine entailing major body organs, including the lung, cardiovascular system, central nervous system, liver, kidney, testes, ovary, pancreas, and esophagus. The mechanism of addiction is also addressed based on interaction of iminiums with normal electron transport chains or electrical phenomena in the brain. The process might occur with or without participation of reactive oxygen species. Evidence indicates that free radicals are widely involved in cell signaling entailing redox processes in the categories of ion transport, neuromodulation, and transcription. Low levels of radicals appear to participate since high concentrations are associated with toxicity. Various possibilities for future work based on the hypothetical approach are addressed, including some that may have practical utility in relation to health improvement, toxicity, and addiction. Insight should be gained from computational studies on the energetics of electron uptake by metabolic iminiums, and on stability of the resultant delocalized radicals. Additional large-scale investigations of antioxidant effects are needed in order to resolve prior conflicting reports. Other proposals are based on interference with metabolism to iminiums and nitrosamines, and destruction of harmful metabolites. Since the iminium entities are proposed to play crucial, adverse roles, it would be worthwhile to explore them with regard to receptors, physiological activities, possible generation of reactive oxygen species, and effect of antioxidants. PMID- 15533624 TI - Dendritic cells and the immunity/tolerance decision. AB - The role of dentritic cells in initiating the immune response has been well established. Recent studies point to an important role for DCs in the induction of peripheral tolerance as well. It was proposed that the role of DC in the immunity/tolerance decision could be associated simply with DC maturation states. However, it has been observed that immature DC do not process endocytosed antigens well to form MHC+peptide complexes and therefore self-specific T cells would not be able to recognize their ligands on immature DCs. Then how might immature DCs induce tolerance to self-antigens? Below it is discussed a new mechanism which might control whether the DCs behavior will be tolerogenic or immunogenic. The hypothesis proposed that DCs should have two maturation programs operating in absence/presence of Danger signals leading to mature-tolerogenic and mature-immunogenic phenotypes, respectively. PMID- 15533625 TI - The meaning of mammalian adult neurogenesis and the function of newly added neurons: the "small-world" network. AB - Adult neurogenesis has been observed in mammalian brain including human but a question remains: how do new neurons become functional in the adult brain? We propose that the random addition of only a few new neurons functions as a maintenance system for the brain's "small-world" networks. In popular parlance, the small-world network phenomenon is described by the concept of "six degrees of separation", which postulates that everyone in the world is connected to everyone else through a chain of at most six mutual acquaintances. Randomly added to an orderly network, new links enhance signal propagation speed and synchronizability. Newly generated neurons are ideally suited to become such links: they are immature, form more new connections compared to mature ones, and their number but not their precise location may be maintained by continuous proliferation and dying off. Similarly, we envisage that the treatment of brain pathologies by cell transplantation would also create new random links in small world networks and that even a small number of successfully incorporated new neurons may be functionally important. PMID- 15533626 TI - Bupropion for fatigue and as a tumor necrosis factor-alpha lowering agent in primary biliary cirrhosis. AB - Primary biliary cirrhosis (PBC) is a chronic cholestatic disease which can often be severe, progressive and necessitate liver transplantation. The cause of PBC is not known, and treatments other than liver transplantation are often not effective. Among the more common and troublesome symptoms of PBC is fatigue. The etiology of fatigue in PBC is not well-understood, and there is no known treatment for it. Here, we suggest that for a number of reasons that the safe and commonly used oral antidepressant bupropion might be effective for fatigue in PBC: (1) increased monoaminergic and dopaminergic tone to combat fatigue, (2) treatment of concomitant depression, (3) in general for PBC as a tumor necrosis factor-alpha (TNF) lowering agent, if TNF is eventually found to play a role in PBC pathogenesis. PMID- 15533627 TI - Flavopiridol: an antitumor drug with potential application in the treatment of neurodegenerative diseases. AB - Several lines of evidence show that cyclin-dependent kinases (CDKs) contribute to neurodegenerative disorders such as Alzheimer's and Parkinson's diseases, and amyotrophic lateral sclerosis. Given their role in the neuronal apoptosis, the inhibition of CDKs by specific drugs such as flavopiridol may be a valid therapeutic approach. Expression of CDKs was observed in rodent models of excitotoxicity and stroke, and CDK inhibitors showed neuroprotective effects. Flavopiridol may provide significant improvement in neurodegenerative diseases in humans. PMID- 15533628 TI - Digoxin may reduce the mortality rates in patients with congestive heart failure. AB - Digoxin has been used to treat congestive heart failure (CHF) for more than two centuries. It's clinical efficacy, however, has been under question in recent years because recent clinical trials showed that digoxin therapy in CHF patients was associated with no beneficial effects in mortality, but only a modest reduction in clinical symptoms and the frequency of heart failure related hospitalisation. Digoxin's effect on mortality seems closely related to its serum concentrations; high serum concentrations (e.g. >or=1.2 ng/ml) have been found to increase the risk of all-cause mortality in heart failure patients. Digoxin associated risk in mortality may be due to an increases in myocardial oxygen consumption and arrhythmogenesis at higher serum concentrations. We hypothesized that the serum concentration of digoxin is a major determinant factor of its efficacy on mortality rates in patients with congestive heart failure. The maintenance of digoxin's serum concentration at the lower end of the reference range, i.e., between 0.5 and 0.8 ng/ml, may reduce mortality rates as well as improve clinical symptoms. PMID- 15533629 TI - On mechanism of superoxide signaling under physiological and pathophysiological conditions. AB - It has been demonstrated that in various physiological and pathophysiological processes superoxide functions as a signaling molecule by the way different from those mediated by hydrogen peroxide, hydroxyl radicals, or peroxynitrite. However, until now the mechanism of superoxide signaling remains obscure. A well known role of superoxide as a precursor of reactive hydroxyl radicals by the superoxide-dependent Fenton reaction or the formation of peroxynitrite must result in the damage of the target molecules and lead to pathological disorders. However, this mechanism is unlikely in such processes as the stimulation by superoxide of enzymatic phosphorylation and dephosphorylation. But, not being a "super-oxidant", superoxide possesses the frequently forgotten "super" nucleophilic properties. Now, we propose a new mechanism for superoxide signaling depending on its nucleophilic reactions. Possible nucleophilic mechanisms of superoxide signaling in the hydrolysis of phosphatidylinositol to inositol 1,4,5 tris-phosphate and in the catalysis of phosphorylation by mitogen-activated protein kinases, phospholipase C and other enzymes are considered. PMID- 15533630 TI - Mammalian sleep may have no adaptive advantage over simple activity-rest cycles. AB - The adaptive value of sleep remains unknown in spite of the intense research performed throughout the last decades. However, few sleep researchers are aware of the difficulties posed by the blind acceptance of an extreme adaptationist viewpoint. Under this philosophy, every anatomical and functional detail present in a living being should have a positive adaptive value, a position that has been considered as rather doubtful. In this report, it is proposed that most of the physiological changes used for mammalian sleep definition could be mere by products of other true adaptations, such as the ontogenetic and phylogenetic development of the nervous system. As a result, complex mammalian sleep could have no adaptive value over that of the simplest forms of rest-activity cycles present in all living forms. In addition, it is proposed that the absence of adaptive value should, by default, be the first option regarding the function of sleep. Besides, the burden of the proof should be always charged over the proponents of every particular adaptive function. As this proof has not been reached, it is the absence of function for sleep which should be taken for granted. PMID- 15533631 TI - Mitochondrial signals to nucleus regulate estrogen-induced cell growth. AB - Classical genomic and non-genomic signaling pathways mediated by nuclear and cell membrane estrogen receptors are considered to contribute to estrogen-induced cell proliferation. Here we propose that mitochondrial signals to the nucleus regulate estrogen-induced progression of the cell cycle. The influence of estrogen on mitochondrial oxidative phosphorylation and mitochondrial gene transcription support the idea that mitochondria are significant targets of estrogen. Mitochondria are the major source of reactive oxygen species (ROS) in epithelial cells. Estrogen redox cycling within mitochondria also generates ROS. Antioxidants inhibit estrogen-induced cell growth. A-Raf, Akt, PKC, MEK, ERK, and transcription factors AP-1, NF-kappaB, and CREB are targets of both estrogen and ROS. We provide four lines of evidence in support of our hypothesis that estrogen induced mitochondrial ROS stimulate redox sensor kinase A-Raf, Akt or PKC, which, in turn, activate transcription factors NF-kappaB, CREB, or AP-1 via the MEK/ERK pathway. Thus, estrogen-induced mitochondrial ROS leading to the activation of cell cycle genes containing AP-1, NF-kappaB, or CREB response elements are involved in the progression of the cell cycle of the estrogen-dependent cells. Our novel concept will contribute to the development of new targets in the prevention and control of estrogen-induced disease including cancer. PMID- 15533632 TI - A "new-old" way of thinking about brain disorder, cerebral excitability--the fundamental property of nervous tissue. AB - Cerebral excitability is normally distributed, and pubertal age is a distinguishing factor. The final developmental event in CNS comprising selective pruning of excitatory synapses coincides with puberty. With early puberty, excess excitation and synaptic density, we have photic susceptibility, paroxysmal EEGs, disturbed circadian rhythms, paroxysmal disorders treated with drugs lowering excitation. Manic-depressive psychosis accords with this. Migraine with paroxysmal EEG, photophobia, hemianopsia, scintillating scotomas, excess excitation in the visual system, benefits from lowering excitation. With late puberty, attenuated CNS, we have disorders in need of raising excitation to avoid breakdown of circuitry, insufficient fill-in mechanism, silent spots, subjectively experienced only--objectively verifiable psychosis: i.e., schizophrenia treated with convulsant neuroleptics. By affecting pubertal age, we affect the distribution of excitation and of post-pubertal brain disorders in accordance with their level of excitation. Excitation is equally important in chronic disorders: l'dopa adversity in Parkinsonism could be due to further lowering of excitation in patients with a deficiency, a schizophrenia-like psychosis develops. Given unavoidable adversity of anti-psychotics, and a marked rise in suicide in schizophrenic and manic-depressive since their introduction, we want to prevent the occurrence of disorders at the extremes, whether very early or late puberty. DHA normalises excitability at all levels of excitation. An adequate daily intake of DHA, before puberty as well as after, might probably reduce or eliminate a development of psychopathology. Lithium is a robust neurotropic agent, and lithiation of the drinking water could be a way of reducing suicide, homicide, violent behaviour, and drug abuse. PMID- 15533633 TI - Nutraceutical resources for diabetes prevention--an update. AB - There is considerable need for safe agents that can reduce risk for diabetes in at-risk subjects. Although certain drugs--including metformin, acarbose, and orlistat--have shown diabetes-preventive activity in large randomized studies, nutraceuticals have potential in this regard as well. Natural agents which slow carbohydrate absorption may mimic the protective effect of acarbose; these include: soluble fiber--most notably glucomannan; chlorogenic acid--likely responsible for reduction in diabetes risk associated with heavy coffee intake; and legume-derived alpha-amylase inhibitors. There does not appear to be a natural lipase inhibitor functionally equivalent to orlistat, although there are poorly documented claims for Cassia nomame extracts. Metformin's efficacy reflects activation of AMP-activated kinase; there is preliminary evidence that certain compounds in barley malt have similar activity, without the side effects associated with metformin. In supraphysiological concentrations, biotin directly activates soluble guanylate cyclase; this implies that, at some sufficient intake, biotin should exert effects on beta cells, the liver, and skeletal muscle that favor good glucose tolerance and maintenance of effective beta cell function. Good magnesium status is associated with reduced diabetes risk and superior insulin sensitivity in recent epidemiology; ample intakes of chromium picolinate appear to promote insulin sensitivity in many individuals and improve glycemic control in some diabetics; calcium/vitamin D may help preserve insulin sensitivity by preventing secondary hyperparathyroidism. Although conjugated linoleic acid--like thiazolidinediones, a PPAR-gamma agonist--has not aided insulin sensitivity in clinical trials, the natural rexinoid phytanic acid exerts thiazolidinedione-like effect in animals and cell cultures, and merits clinical examination. Other natural agents with the potential to treat and possibly prevent diabetes include extracts of bitter melon and of cinnamon. Nutraceuticals featuring meaningful doses of combinations of these agents would likely have substantial diabetes-preventive efficacy, and presumably could be marketed legally as aids to good glucose tolerance and insulin sensitivity. PMID- 15533634 TI - The putative therapeutic value of high-dose selenium in proliferative retinopathies may reflect down-regulation of VEGF production by the hypoxic retina. AB - Two decades ago, Crary reported his anecdotal observation that a supplemental antioxidant regimen including high-dose selenium (500 mcg daily) appeared to slow the progression of visual loss in diabetic retinopathy and macular degeneration. Recently, selenium has been shown to down-regulate VEGF production by rodent cancer cells, both in vivo and in vitro; this effect is dose-dependent and, like the anticarcinogenic activity of selenium, is mediated by methyselenol. If increased selenium exposure can likewise suppress VEGF production in the hypoxic retina, a straightforward explanation for Crary's observation may be at hand. Since selenium is inexpensive and non-toxic in the dose employed by Crary, an effort to replicate his findings is warranted. PMID- 15533635 TI - The role of lipoprotein (a) in pregnancies complicated by pre-eclampsia. AB - Endothelial cell dysfunction is a key feature of the pathogenesis of pre eclampsia. The cause of the endothelial cell injury is probably multifactorial, but poor placenta perfusion plays a major role. In pre-eclampsia, characteristic pathological lesions in the placenta are fibrin deposits, acute atherosis and thrombosis. The similarity between the lesions of pre-eclampsia and atherosclerosis has led to speculations of a common pathophysiological pathway. An abnormal lipid profile is known to be strongly associated with atherosclerotic cardiovascular disease and has a direct effect on endothelial function. Abnormal lipid metabolism seems important in the pathogenesis of pre-eclampsia too. An elevated plasma lipoprotein (a) concentration is a known risk factor for atherosclerotic cardiovascular disease. In this paper, we discuss three hypotheses about the mechanisms by which lipoprotein (a) may be associated with pre-eclampsia: 1. Lp(a), as an acute-phase reactant, transporting cholesterol to sites of endothelial damage for reparation, temporarily increases during pregnancy and increases more during a pregnancy complicated by mild to moderate pre-eclampsia as compared to an uncomplicated pregnancy, in response to a greater extend of endothelial injury in pre-eclampsia. After delivery, pre-eclampsia subsides and Lp(a) concentrations return to baseline levels. 2. In cases of severe pre-eclampsia, there is even more extensive endothelial damage and consequently a higher consumption of Lp(a) in reparation of this vascular damage. These women will have lower concentrations of Lp(a). 3. High baseline concentrations of Lp(a), which are genetically determined, may induce or contribute to the development of pre-eclampsia by promoting endothelial dysfunction. In this line of reasoning one would expect to find higher concentrations of Lp(a) in women at risk for developing pre-eclampsia in a future pregnancy or with a history of pre-eclampsia. As discussed above, these women are also at increased risk for future cardiovascular disease as compared to women with a history of normal pregnancy. The pathophysiologic changes associated with cardiovascular disease may also be responsible for the increased incidence of pre eclampsia in these women. PMID- 15533636 TI - Accidentally created tension pneumothorax in patient with primary spontaneous pneumothorax--confirmation of the experimental studies, putting into question the classical explanation. AB - BACKGROUND: The widespread explanation of patophysiology of tension pneumothorax is that compression to the mediastinum by the progressively accumulating intrapleural air causes torsion at the atrio-caval junction, impaired filling of the right heart and circulatory arrest as potentially life-threatening complication. Some experimental studies on animals put into question such an explanation, suggesting that respiratory arrest due to hypoxia of the respiratory center, not a circulatory arrest, represents dominant life threatening feature. CASE REPORT: we present a patient with spontaneous pneumothorax in whom tension pneumothorax occurred accidentally, i.e., in whom air was insufflated under great pressure from the aspirating system into the pleural cavity, immediately after insertion of a chest tube. As the situation was recognized immediately, urgent reanimation was undertaken--endotracheal intubation, ventilation through the balloon, reconnection of the chest tube to another aspirating system. Lung reexpansion was achieved and the patient was discharged after an uneventful course. In this patient, it was possible to register the sequence of events before, during and after the incident. Dominant clinical finding during resuscitation of this apnoic, cyanotic and unconscious patient was respiratory arrest in presence of evident maintenance of peripheral circulation, that supports results of experimental studies. Dominant findings in experiments with creation of tension pneumothorax was that, although pressures rose throughout the right side of the circulation, no developing pressure gradient was found on this side of the circulation; furthermore, respiratory arrest preceded cardiac arrest in these animals. Hypoxia of the respiratory center, caused by the increasing portion of pulmonary blood flow being shunted through nonventilated or hypoventilated lung, was suggested as primary cause of death of experimental animals. The same factor seems to be a cause of respiratory arrest in our patient. CONCLUSION: respiratory arrest, preceding circulatory arrest, seems to be the principal life threatening condition in patients with progressive tension pneumothorax. PMID- 15533638 TI - Pleomorphic mammalian tumor-derived bacteria self-organize as multicellular mammalian eukaryotic-like organisms: morphogenetic properties in vitro, possible origins, and possible roles in mammalian 'tumor ecologies'. AB - Highly pleomorphic bacteria have regularly been isolated from mammalian tumors and leukemic bloods. Here, it is shown that highly pleomorphic, cell-wall deficient bacteria derived from a mammalian tumor self-organize in vitro into mammalian tissue-like morphogenetic patterns consisting of multicellular tissue like sheets and capillary-like networks. It is proposed that these pleomorphic mammalian tumor-derived (MTD) bacteria, during morphogenesis, express mammalian tissue morphogenesis-related genes that were acquired through eukaryote-to prokaryote DNA transfer. Similar pleomorphic MTD bacteria might play important roles as symbiotic multicellular mammalian eukaryotic-like organisms in mammalian 'tumor ecologies' that include malignant and nonmalignant mammalian eukaryotic cells. From a mammalian tumor ecology perspective, eradication of tumors in some mammalian hosts may depend upon the elimination of pleomorphic MTD bacteria self organized as symbiotic multicellular mammalian eukaryotic-like organisms. Further investigations of the extraordinary mammalian eukaryotic-like multicellularity of these bacteria may yield fundamental insights into the evolution of multicellularity and multicellular development and may challenge basic assumptions regarding cellular evolution. PMID- 15533637 TI - Immunosuppressive therapy for kidney transplant prevents vaso-occlusive crisis in a haemoglobin SC disease patient. AB - Although the molecular basis of sickle cell disease (SCD) is well established, the wide variability in clinical manifestations still puzzles haematologists and clinicians. Recently, SCD started to be considered by different groups as a chronic inflammatory condition, where the inflammatory tendency of each individual could drive more or less severe clinical features. Here we describe a haemoglobin SC disease patient (heterozygous to both HbS and HbC variants) that experienced several vaso-occlusive crises before underwent a successful kidney transplantation. Since then (16 years ago), she is on uninterruped immunosuppressive therapy, and do not experienced any severe vaso-occlusive crisis. Considering SCD associated morbidity as a result of exacerbated immune responses, we suggest that the immunosuppressive therapy directed to the kidney graft maintenance is actually also helping in the control of the chronic inflammatory responses associated to SCD. PMID- 15533639 TI - Abnormal proinsulin congeners as autoantigens that initiate the pathogenesis of Type 1 diabetes. AB - Exposure of concentrated and purified monomeric insulin solutions to inorganic oxidants as iodine and chlorine lead to the appearance of a minor peak on gel chromatography that was disulfide cross-linked. It exhibited a 20-fold reduction in bioactivity, a markedly decreased immunoreactivity and a different electrophoretic pattern as compared to the starting material. The covalent dimer was formed by way of aggregation. These findings were reproduced by incubating monomeric insulin in normal blood spiked to hyperglycemic levels. In contrast, normoglycemic blood used as incubating medium failed to induce dimerization. Since the conformation of proinsulin is similar to that of insulin, involving the exposure of the anterior A7-B7 disulfide bridge, the authors hypothesize that proinsulin dimers rather than insulin dimers might be formed in Type 1 diabetes (TD1), leading to the autoimmune destruction of pancreatic B-cells. Proinsulin is present in a soluble aggregate state in the coated granule and may further accumulate allowing disulfide exchange due to abnormalities of the processing enzymes. This might be caused by inborn errors in genetically susceptible children or perhaps secondary to viral infection, whereas crystallization of insulin in mature granules is unlikely to be conducive to dimerization. Dimeric proinsulin would then migrate to pancreatic B-cell membranes to be taken up by surface immunoglobulins on B-lymphocytes that would in turn present it to cytotoxic T lymphocytes. The abnormal configuration of this congener would not be recognized as self by the immune system, triggering a selective destruction of pancreatic B-cells in the early pre-clinical development of TD1, resulting eventually in clinical disease. Since proinsulin is continuously converted to insulin in the coated granules of Type 2 diabetics, dimerization is unlikely in this condition. As pointed out by earlier investigators, TD1 is not a homogeneous disease, since several of its clinical features are different in children up to 6 years of age as compared to older patients. It is thus conceivable that there are subsets of TD1 triggered by dissimilar autoantigens and we propose in the present hypothesis that a conformationally altered congener of native proinsulin might play such a role. PMID- 15533640 TI - Statin, like aspirin, should be given as early as possible in patients with acute coronary syndrome. AB - It is estimated that about 1 million patients are hospitalized for acute coronary events each years in the United States. An acceptable theory is that the acute coronary syndrome is caused by rupture of the atherosclerotic plaque with superimposed thrombus, which is a complex process and involving a number of different stages. Previous studies indicated that inflammation is one of the most important features of vulnerable plaque, and occurs in most vulnerable plaque, comprised of monocytes, macrophages, and lymphocytes in both the cap and in the adventitia. This is supported by evidence that reduction in serum inflammatory marker levels, such as C-reactive protein, significantly decreased coronary events in patients with acute coronary syndrome. A large number of investigations have demonstrated that administration of statin could modify C-reactive protein concentrations with a concurrent fall in cardiovascular events. Our recent data indicated that reduction of inflammatory markers could be achieved within 24 h following a single dose of statin administration after admission in patients with coronary artery disease. Based on the available evidence and in light of the new understanding that statins have pleiotropic effects, especially as a potent anti inflammatory agent, the statins, like aspirin, should be clinically given as early as possible in patients with acute coronary syndrome. PMID- 15533641 TI - Adenosine A2a receptor/dopamine D2 receptor hetero-oligomerization: a hypothesis that may explain behavioral sensitization to psychostimulants and schizophrenia. AB - The mechanisms underlying psychostimulant-induced behavioral sensitization and schizophrenia are yet to be fully elucidated. Evidence suggests that the dopamine D2 receptor (DRD2) as well as other neurotransmitter system receptors may be involved in the two conditions, and previous reports have hypothesized that oligomerization of dopamine receptors and analogs from other neurotransmitter systems may underlie the mechanism responsible. This paper describes findings which show co-localization of DRD2 and the adenosine A2a receptor (A2aAR) in the striatum, interaction between the two receptors and A2aAR/DRD2 hetero oligomerization in the neuronal cell. The possibility that A2aAR/DRD2 hetero oligomerization may be involved in the development of psychostimulant-induced behavioral sensitization or the pathogenesis of schizophrenia is explored. A2aAR/DRD2 hetero-oligomerization can enhance dopaminergic function, resulting in an increased behavioral response to psychostimulants or the development of psychotic symptoms. Receptor oligomerization can affect receptor degradation, which may affect the persistence of behavioral sensitization or psychotic symptoms. The proposed A2aAR/DRD2 hetero-oligomerization model may suggest new directions in the understanding and treatment of schizophrenia. PMID- 15533642 TI - Aging: gene silencing or gene activation? AB - According to the author's theory of gene silencing, the key process in aging involves reduced expression of a number of genes. Silencing of genes has a complex mechanism, which involves methylation of DNA, histone modification and chromatin remodeling. In addition to deacetylation of the histones and methylation of DNA, recently described RNAi mechanism could initiate formation of silenced chromatin. Hypermethylation of the promoter will silence the gene. Genome-wide hypomethylation will induce genomic instability, amplification of oncogenes and also silencing of the genes through RNAi mechanism. Studies by different groups, conducted in yeast, worms, flies and mice, confirmed substantial changes in gene expression in aging. Among them, the most important was silencing of tumor suppressors and other genes involved in the control of cell cycle, apoptosis, detoxification, and cholesterol metabolism. There was also increased expression of the smaller group of oncogenes and other genes which are associated with typical diseases of old age. Caloric restriction normalizes expression of a substantial percentage of these genes. Animal studies confirmed importance of caloric restriction, which decreases signaling through the IGF 1/AKT pathway and expression of gene p53. These studies, however, cannot be directly applied to human aging. It is proposed that age management therapy should attempt to normalize gene expression in the older population to the level typical for young adults. This would require activation of silenced genes and normalization of overexpressed genes. Caloric restriction and exercise are helpful in decreasing the activity of important oncogenes and activation of silenced tumor suppressors, and may have a positive impact, not only on aging, but also on prevention of cancer. Dietary supplements containing phytochemicals should normalize increased expression of oncogenes. Examples are: genistein and EGCG, which effect signaling through the IGF-1/AKT pathway and resveratrol and limonen, which do so through the RAS pathway. A group of amino acid derivatives and organic acids of animal and human origin should activate silenced tumor suppressor genes (Aminocare A10, Aminocare Extra). Among them 3-phenylacetylamino 2, 6-piperidinedione intercalates specifically with DNA and protects sequences of tumor suppressor genes, which are vulnerable to the effects of carcinogens. Phenylacetate activates p53 and p21 through inhibition of methyltransferase and farnesylation of the RAS protein. Phenylbutyrate activates tumor suppressor genes through inhibition of histone deacetylation. Phenylacetylglutamine decreases genomic instability and expression of oncogenes and promotes apoptosis. The application of DNA microarray techniques to human studies should provide more information about differences in gene expression in different age groups and help design more effective age management regimens. PMID- 15533643 TI - Two-plane symmetry in the structural organization of man. AB - Manifestations of symmetry in the human structural organization in ontogenesis and phylogenetic development are analysed. A concept of macrobiocrystalloid with inherent complex symmetry is proposed for the description of the human organism in its integrity. The symmetry can be characterized as two-plane radial (quadrilateral), where the planar symmetry is predominant while the layout of organs of radial symmetry is subordinated to it. Out of the two planes of symmetry (sagittal and horizontal), the sagittal plane is predominant: (a) the location of the organs is governed by two principles: in compliance with the symmetry planes and in compliance with the radial symmetry around cavities; (b) the location of the radial symmetry organs is also governed by the principle of two-plane symmetry; (c) out of the four antimeres of two-plane symmetry, two are paired while the other two have merged into one organ; (d) some organs which are antimeres relative to the horizontal plane are located at the cranial end of the organism (sensory organs, cerebrum-cerebellum, heart-spleen and others). The two plane symmetry is formed by two mechanisms--(a) the impact of morphogenetic fields of the whole crystalloid organism during embriogenesis and (b) genetic mechanisms of the development of chromosomes having two-plane symmetry. When comparing mineral and biological entities we should consider not the whole immobile crystal but only the active superficial part of a growing or dissolving crystal, the interface between the crystal surface and the crystal-forming environment which directly controls crystal growth and adapts itself to it, as well as crystal feed stock expressed in the structure of concentration flows. The symmetry of the chromosome, of the embrion at the early stages of cell cleavage as well as of some organs and systems in their phylogenetic development is described. PMID- 15533644 TI - Epialleles and common disease. PMID- 15533645 TI - Could changing the ideal anthropometric values of children help prevent starvation on Earth. PMID- 15533646 TI - Sudden death in sickle cell trait: could coexistent diabetes play a role? PMID- 15533647 TI - Vasopressin: a pre-requiste for some actions of renal nitric oxide? PMID- 15533648 TI - ECT treatment for autism: a viable alternative for a subgroup of patients? PMID- 15533649 TI - Founders of child neurology in Japan--Munenori Enjoji. PMID- 15533650 TI - Recent advances in the biochemistry and genetics of sphingolipidoses. AB - Sphingolipidoses are a subgroup of lysosomal storage diseases. They are defined as disorders caused by a genetic defect in catabolism of sphingosine-containing lipids. Catabolism of these lipids involves enzymes and activator proteins. After the discovery of lysosomes by de Duve and the demonstration of the first defective lysosomal enzyme by Hers in 1963, the first enzyme deficiency for sphingolipidoses was characterized in 1965 and all the defective enzymes were demonstrated in the last three decades. In 1984, the first activator protein was found and it expanded the concept of sphingolipidoses. In the following years, many researches have been undertaken to understand the molecular basis of these diseases, the mechanism of pathogenesis, the mechanism of lysosomal digestion of glycosphingolipids (GSLs) and the functional domains of lysosomal enzymes. New hypotheses and theories have been put forward for the mechanism of lysosomal digestion and pathogenesis. However, although much has been done, the pathogenesis of sphingolipidoses has not been fully elucidated. Mouse models of these diseases have facilitated the elucidation of pathogenesis and the development of therapeutic strategies for these diseases, which are not treatable at present except for Fabry and type 1 Gaucher disease. The purpose of this review is to collect information on the recent researches related to sphingolipidoses. The review includes the hydrolysis of GSLs in lysosome, mechanism of hydrolysis, pathogenesis and genetics of sphingolipidoses, a brief mouse model and therapeutic strategies of these diseases. PMID- 15533651 TI - Visual-perceptual impairment in children with periventricular leukomalacia. AB - We set out to define visuo-perceptual impairment related to periventricular leukomalacia (PVL) using the Developmental Test of Visual Perception (DTVP). Correlations were sought between visual-perceptual deficits and DTVP profile and neuroradiological and neurophthalmological findings. The DTVP was administered to 20 children (m/f: 10/10), aged between 5 and 8 years (mean: 6.95 years), presenting with: spastic diplegia; PVL documented by brain MRI; normal or mildly impaired visual acuity; mild-moderate upper limb functional impairment. The mean General Visual-Perceptual Quotient was impaired, showing a great variability among the patients. Despite this, an uneven DTPV profile, characterised by a significant difference between the VMIQ and the Non-Motor Visual-Perceptual Quotient (P < 0.001) and a poor result on the Closure subtest (identification of whole figures from incomplete visual information) was observed in all the subjects. This profile reflects a deficit in eye-hand coordination and in praxic constructional abilities and could be the expression of malfunctioning of the occipital-parietal pathway of visual integration, the so-called 'dorsal stream,' a hypothesis reinforced by the emergence of a statistically significant correlation between the neuroradiological data and the presence of visual perceptual impairment. PMID- 15533652 TI - EEG spectral analysis in children with febrile delirium. AB - Febrile delirium is defined as an acute and transient confusional state with high fever. There are very few reports on febrile delirium, although fever is one of the commonest symptoms in children. We previously found a posterior slowing in the electroencephalogram (EEG) of delirious patients with fever. The purpose of this study is to evaluate the features of occipital slow waves by spectral analysis and to find a parameter associated with clinical improvement. METHODS: Digital EEG tracings were investigated by Fourier analysis in 20 patients aged from 2 to 13 years. The fast Fourier transform (FFT) was computed for 20 s tracing from the P3-A1 and P4-A2 derivations. The spectral analysis of EEG was repeated in 7 patients. The tracings of 34 control subjects were also analyzed by FFT. EEG of a febrile, nine-year-old girl without delirium was also studied. RESULTS: Febrile delirium was seen during the first three days of fever. The episodes lasted up to 10 min. Four patients showed febrile delirium again after admission but they became conscious a few minutes later. The relative power in the delta frequency band was increased in 65% of patients with preservation of the occipital alpha rhythm. In addition, repeated febrile delirium did not cause worsening of the posterior slowing. The duration of abnormal EEG was only a few days and the decrease of relative power in the delta frequency band was the best parameter of clinical improvement. Posterior slowing was also found in a febrile patient without delirium. CONCLUSION: Febrile delirious children showed the characteristic clinical and spectral analytical features and the numerical data of EEG facilitate the comparison of the serial findings. PMID- 15533653 TI - Discordant mental and physical efforts in an autistic patient. AB - We investigated whether there was mental effort in response to verbal commands in a 16-year old girl with autism, a high degree of non-compliance with commands and symptoms of autonomic dysfunction by monitoring the brainstem autonomic tone during an attempt to perform isometric exercise. An index of cardiac vagal tone (CVT), cardiac sensitivity to baroreflex (CSB), heart rate (HR) and mean arterial blood pressure (MAP) were measured simultaneously. Physical non-compliance with our commands meant there was no force applied by the patient during the attempted exercise, but CVT and CSB were both reduced and sustained at very low levels throughout the attempt, while MAP and HR were increased concurrently to higher levels in the same period. This vagal withdrawal to allow concurrent increases in HR and MAP is an arousal response appropriate for isometric exercise, which is a sign of a positive mental effort to comply with our commands. These results demonstrate discordant mental and physical efforts in our patient. In this particular case, the physical inabilities in some instances could have been mislabelled as mental non-compliance due to autism. It would be worthwhile to investigate the prevalence of discordant mental and physical efforts in autism. PMID- 15533654 TI - A pilot study on lidocaine tape therapy for convulsions with mild gastroenteritis. AB - The aim of this study was to clarify the efficacy and safety of lidocaine tape therapy (LDT) in patients with convulsions with mild gastroenteritis (CwG). Twenty-one consecutive episodes of CwG were treated with LDT therapy. The dose of LDT was 36 mg in patients with body weights of <15 kg, 54 mg in those with body weights between 15 and 20 kg, and 72 mg in those with body weights of >20 kg. LDT was attached on the back of each patient every 12 h. Application of LDT was continued for 48 h. The serum levels of lidocaine (LD) were measured 4 h after the first attachment of LDT. The seizures completely ceased in 13 episodes (62%) after the application of LDT, while a recurrence of seizures was observed in the other 8 episodes. Drip infusion of LD was performed in 4 of these 8 episodes. The serum LD levels were only measurable in 6 infants (average, 0.4 microg/ml; range, 0.2-0.5 microg/ml). In the other patients, the serum LD levels were below the lower limit of measurement. Adverse effects of LDT were not observed in any patients. LDT therapy was safe and effective in patients with CwG. PMID- 15533655 TI - A major influence of CYP2C19 genotype on the steady-state concentration of N desmethylclobazam. AB - N-desmethylclobazam (N-CLB), the major metabolite of clobazam (CLB), exerts a large influence on therapeutic and adverse effects of CLB. A substantial inter individual variability has been observed in the ratios of N-CLB concentration/CLB dose and of the N-CLB/CLB concentration. We document here a genotype-phenotype correlation between CYP2C19 polymorphisms and those ratios. Patients with two mutated CYP2C19 alleles show significantly higher ratios than those with the wild type genotype: patients with one mutated allele exhibited intermediate trait. That is, the degree of elevation in the ratios was dependent on the number of mutated alleles of CYP2C19 (gene-dose effect). The N-CLB concentration/CLB dose ratio of patients with two mutated alleles was more than six fold higher than that of wild type patients. Thus, the serum N-CLB/CLB concentration ratio may be a valuable parameter to screen for patients at risk for side effects. Such precautions may be clinically relevant in populations where the mutant allele frequency is high, such as in Asian populations ( approximately 35%). Patients co medicated with CYP3A4 inducer showed lower CLB concentration/CLB dose ratios and higher N-CLB/CLB concentration ratios. The overall effect of CYP3A4 inducer on N CLB metabolism, however, was small and, thus, we conclude that the CYP2C19 genotype is the major determinant of the N-CLB concentration. For this reason it is crucial for the better management of epilepsy and other chronic illnesses in general to establish the correlation of genotype of CYP enzymes and pharmacokinetics/dynamics of drugs. PMID- 15533656 TI - MR imaging and 1H-MR spectroscopy in a case of cerebral infarction with transient cerebral arteriopathy. AB - A case of cerebral infarction with transient cerebral arteriopathy in a 12-year old female is described. The child showed with an acute onset of left hemiplegia, central facial palsy and aphasia. She was diagnosed with right internal carotid artery obstruction with no predisposing factors for cerebral infarction. A serial MR image and MR spectroscopy of the brain was performed. MR angiography revealed reversible vascular changes in the right middle cerebral artery after several months. The ratio of choline/creatine was elevated and that of N-acetylaspartate (NAA)/creatine was depressed in the cerebral lesion, however, they were normalized during the remission phase with long echo time-MR spectroscopy. The elevation of the lactate peak was clarified with short echo time-MR spectroscopy. The longitudinal follow up using MR angiography, and MR spectroscopy with long and short echo time was useful for the evaluation of reversible vascular change with cerebral infarction in children. PMID- 15533657 TI - Dystonia in a 13-year-old boy with secondary progressive multiple sclerosis. AB - We report a patient who developed relapsing-remitting multiple sclerosis (MS) at 8 years old, and then had a progressive clinical course and dystonia. Dystonia of the patient is probably due to a lesion of the basal ganglia. Abnormal posture or movement disorder is very rarely found in MS, and progressive clinical course is also rare in childhood. The patient is worthy of attention because of his childhood onset, progressive clinical course and dystonia. PMID- 15533658 TI - Acute dysautonomia: complete recovery after two courses of IVIg. AB - An 11-year-old boy presented with acute dysautonomia manifesting as severe orthostatic hypotension following fever. Serial orthostatic tests with measurement of the coefficient of variation in the R-R intervals showed improvement after one course and complete recovery after two courses of intravenous high-dose immunoglobulin therapy (IVIg). Repeated courses of IVIg should be considered to treat this disorder if spontaneous remission does not occur. PMID- 15533659 TI - An analytical perspective on favoured synthetic routes to the psychoactive tryptamines. AB - Many tryptamine derivatives are known to induce altered states of consciousness and are increasingly of interest in forensic and neurobiological studies. The analytical chemistry of certain synthetic routes to the tryptamines is discussed and likely side products and impurities identified, where literature reports are available. Recent examples from the authors' laboratory are presented to highlight future prospects and implications for analytical procedures. The aim of this review is to provide the analytical chemist with the foundation chemistry and some analytical targets to be able to undertake direct characterisation of products and intermediates. These might become available from interdiction of clandestine operations in a forensic environment or during the synthesis of the tryptamines for investigative neurobiological and clinical procedures. PMID- 15533660 TI - FI-chemiluminometric study of thiazides by on-line photochemical reaction. AB - The present manuscript deals with a simple and sensitive flow-injection method for the chemiluminescent determination of thiazides. The method is based on the on-line photodegradation and chemiluminescent determination of the resulting photo-fragments. The on-line photodegradation is performed in basic medium by using a photoreactor consisting of a 550cm long x 0.8mm ID piece of PTFE tubing helically coiled around an 8W low-pressure mercury lamp. The determination of the photo-irradiated thiazides is performed by a chemiluminescent oxidative reaction with Ce(IV) in sulphuric acid medium. A heterogeneous group of thiazides (indapamide, metolazone, hydroflumethiazide, chlorthalidone and bendroflumethiazide) has been studied. Hydrochlorothiazide was selected as a test substance. The "on-line" photochemical reaction approach allows the sensitive chemiluminescent determination of thiazides which do not present native chemiluminescence in the absence of sensitizers such as Rhodamine 6G. Linear calibration graphs were typically over the range 0.5-12 microgml(-1) (indapamide, metolazone, hydroflumethiazide and chlorthalidone); and over the range 0.5-5 microgml(-1) (hydrochlorothiazide). Limits of detection ranged between 0.005 microgml(-1) (hydrochlorothiazide) and 0.06 microgml(-1) (bendroflumethiazide). The relative standard deviation for the test substance was 2.0% for 2 microgl(-1) of the drug (n = 11) and the throughput was 65 h(-1) in all cases. The assessment of the photodegradation step on the molecular structure of thiazides was established by recording UV and fluorimetric spectra. The viability of the on line photoinduced fluorescent determination of hydroflumethiazide and bendroflumethiazide was confirmed. The method was also applied to the determination of hydrochlorothiazide in commercially available formulation. PMID- 15533661 TI - Ion selective electrodes for penicillin-G based on Mn(III)TPP-Cl and their application in pharmaceutical formulations control by sequential injection analysis. AB - The work describes the construction, evaluation and analytical application of ion selective electrodes sensitive to penicillin-G antibiotics for pharmaceutical products analysis. Different types of polymeric membranes based on PVC (poly(vinyl chloride)) and EVA (ethyl-vinyl-acetate), without internal reference solution, were prepared using 5,10,15,20-tetraphenylporphyrinate (TPP) manganese(III) (Mn(III)TPP-Cl) as electroactive material. Different additives such as tetra-n-octylammoniumbromide (cationic additive) and sodium tetraphenylborate (anionic additive) were incorporated into the membranes to evaluate their influence on electrodes performance. The comparison of the developed detectors was based on general analytical characteristics, selectivity and lifetime. To accomplish the analysis of real samples, two selective membranes composed of 33.0% (w/w) of PVC, 66.0% (w/w) of o-NPOE and 1.0% (w/w) of Mn(III)TPP-Cl (type A) and 33.0% (w/w) of PVC, 66.0% (w/w) of o-NPOE, 1.0% (w/w) of Mn(III)TPP-Cl and 10% mol (relative to the molar concentration of Mn(III)TPP Cl) of sodium tetraphenylborate (type B) were used. Type A electrode presented a linear response between 2 x 10(-5) and 10(-1) moll(-1) for penicillin-G, a slope of about -59 mVdec(-1) and a reproducibility of about +/-0.5 mVday(-1), while type B exhibited a linear response between 5 x 10(-5) and 10(-1) moll(-1) for penicillin-G, a slope of about -61 mVdec(-1) and a reproducibility of about +/ 0.3 mV day(-1). The potentiometric analysis of penicillin-G in pharmaceutical products was carried out by direct potentiometry and the results obtained were compared with those provided by the HPLC reference method. These membranes (type A and type B) were used to prepare tubular electrodes that were coupled to a sequential injection system (SIA) and presented a linear range between 2 x 10(-4) and 1 x 10(-2) moll(-1) and slopes of -59.3 +/- 0.8 and -57.3 +/- 1.2 mVdec(-1), respectively. The tubular electrode constructed using type B membrane (type TB) was used to carry out the potentiometric analysis of penicillin-G in pharmaceutical formulations. The proposed procedure enabled relative errors between 0.1% and 1.2% (n = 4) and a sampling-rate of about 25 samples per h. PMID- 15533662 TI - Plasmon-waveguide resonance spectroscopy applied to three potential drug targets: cyclooxygenase-2, hepatitis C virus RNA polymerase and integrin alpha V beta 3. AB - Plasmon-waveguide resonance (PWR) spectroscopy has been used to study the interactions between ligands that correspond to inhibitors, activators or substrates and three integral membrane proteins representing potential drug targets; cyclooxygenases 1 and 2 (COX-1 and -2), integrin alphaVbeta3, and hepatitis C virus RNA polymerase. The proteins were incorporated into an egg phosphatidylcholine bilayer deposited onto the surface of the PWR resonator, and changes in the amplitude and position of the PWR spectra due to mass density increases and conformational transitions have been used to characterize the kinetics and binding affinities corresponding to these interactions. Although the partition of COX-2 into the bilayer was not as efficient as was the case with the other two proteins, sufficient protein could be incorporated to allow ligand binding to be observed. It was also possible to incorporate COX-1 into a lipid bilayer by adding a suspension of microsomal membrane fragments containing this enzyme to a preformed bilayer, and to observe binding of an inhibitory ligand. The interactions between integrin alphaVbeta3 and two ligands with different in vivo efficacies could be distinguished by both spectral measurements and binding kinetics. In the case of the RNA polymerase, the kinetics of PWR spectral changes upon adding a substrate solution to an enzyme-template complex were consistent with those obtained from direct measurements of enzymatic turnover. These experiments demonstrate the utility of PWR spectroscopy to provide novel information regarding drug interactions with membrane proteins in a lipid environment and to distinguish conformational changes induced by binding of various drug molecules. PMID- 15533663 TI - Simultaneous determination of dysprosium and iron in urine by capillary zone electrophoresis coupled to cloud point extraction. AB - Automated preconcentration strategies are needed when analyzing metals in real samples by capillary electrophoresis (CE) with UV detection. The on-line incorporation of cloud point extraction (CPE) to flow injection analysis (FIA) associated with CE for simultaneously determining dysprosium and iron at ppb levels in urine is presented and evaluated for the first time. The preconcentration step is mediated by micelles of the non-ionic surfactant polyethyleneglycol-mono-p-nonylphenylether (PONPE 7.5) with 2-(5-bromo-2 pyridylazo)-5-diethylaminophenol. The micellar system containing the complex was loaded into the FIA manifold at a flow rate of 8 mlmin(-1), and the surfactant rich-phase was retained in a microcolumn packed with cotton, at pH 9.2. The surfactant-rich phase was eluted with 50 microl acetonitrile directly into the CE sample vial, allowing to reach an enrichment factor of 200-fold for a 10 ml sample urine. The type and composition of the background electrolytes (BGE) were investigated with respect to separation selectivity, reproducibility and stability. A BGE of 20mM sodium tetraborate buffer containing 13% acetonitrile, pH 9.0 was found to be optimal for the separation of metal chelates. Detection was performed at 585 nm. An enhancement factor of 200 was obtained for the preconcentration of 10 ml of sample solution. The detection limits for the preconcentration of 10 ml of urine were 0.20 microgl(-1) for Dy, and. 0.48 microgl(-1) for Fe. The calibration graphs using the preconcentration system were linear with a correlation coefficient of 0.9989 (Dy) and 0.9976 (Fe) at levels near the detection limits up to at least 500 microgl(-1). The method was successfully applied to the determination of dysprosium and iron in urine for monitoring the elimination of dysprosium-based pharmaceuticals. PMID- 15533664 TI - Separation and determination of synthetic impurities of difloxacin by reversed phase high-performance liquid chromatography. AB - A simple and rapid reversed-phase high-performance liquid chromatographic method for separation and determination of process-related impurities of difloxacin (DFL) was developed. The separation was achieved on a reversed-phase C(18) column using methanol-water-acetic acid (78:21.9:0.1, v/v/v) as a mobile solvent at a flow rate of 1.0 ml/min at 28 degrees C using UV detection at 230 nm. It was linear over a range of 0.03 x 10(-6) to 1.60 x 10(-6)g for process related impurities and 0.05 x 10(-6) to 2.40 x 10(-6)g for difloxacin. The detection limits were 0.009 x 10(-6) to 0.024 x 10(-6)g for all the compounds examined. The recoveries were found to be in the range of 97.6-102.0% for impurities as well as difloxacin. The precision and robustness of the method were evaluated. It was used for not only quality assurance, but also monitoring the synthetic reactions involved in the process development work of difloxacin. The method was found to be specific, precise and reliable for the determination of unreacted levels of raw materials, intermediates in the reaction mixtures and the finished products of difloxacin. PMID- 15533665 TI - Assay for the simultaneous determination of acetaminophen-caffeine-butalbital in human serum using a monolithic column. AB - A fast and sensitive high performance liquid chromatography (HPLC) assay was developed on a C18 monolithic column for the simultaneous determination of acetaminophen-caffeine-butalbital in human serum. Serum samples were treated with a solid phase extraction procedure. The analytes were separated using a mobile phase of 95:5 (v/v) 0.1M potassium phosphate monobasic (pH 2.41)-acetonitrile on the C18 monolithic column with detection at 220 nm. Benzoic acid was used as the internal standard (IS). The method was validated over the range of 1.25-100 microg/ml for each drug and found to be linear (r > 0.995, n = 12) with RSD less than 8.3%. The method proved to be accurate (percent bias for all calibration samples varied from -14.6 to -1.3%) and precise (ranged from 2.9 to 13.4%). The mean percent absolute recoveries from serum were 89.7 +/- 3.6 for acetaminophen, 95.5 +/- 4.5 for caffeine, 99 +/- 5.2 for butalbital and 83.4 +/- 3.9% for the internal standard. PMID- 15533666 TI - Direct determination of diclofenac in pharmaceutical formulations containing B vitamins by using UV spectrophotometry and partial least squares regression. AB - This work proposes a simple and rapid analytical procedure for determination of diclofenac (DCF) in the presence of B vitamins, based on UV measurements and partial least squares (PLS). The interference of thiamine (THI) and pyridoxine (PYR) were modelled using an experimental design constructed in the ranges of 10 50 micromoll(-1) for DCF and THI and 15-75 micromoll(-1) for PYR. The procedure was repeated at five different pH values (between 3 and 6) and the best results were observed at pH 5, presenting a root mean square error of prediction (RMSEP) of 0.80 micromoll(-1) for DCF. The procedure was successfully applied to simultaneous determination of DCF, THI and PYR in synthetic mixtures and in a pharmaceutical formulation that contains a simple excipient (lactose). For determination of a more complex formulation that contains 15 different substances in the excipient, including some UV absorbing ones, the procedure was only able to determine DCF, since the excipient interferences disturbed THI and PYR predictions. Figures of merit, such as selectivity, analytical sensitivity, limit of detection and precision were determined for the DCF prediction model and the determinations were verified by an independent method, HPLC. PMID- 15533667 TI - Development of a liquid chromatographic method for ear drops containing neomycin sulphate, polymyxin B sulphate and dexamethasone sodium phosphate. AB - Two liquid chromatographic methods were developed to analyse ear drops containing neomycin sulphate, polymyxin B sulphate and dexamethasone sodium phosphate. This formulation will be described in the Belgian National Formulary. Since neomycin, an aminoglycoside antibiotic, has no UV chromophore and pre or post column derivatization is complicated, pulsed electrochemical detection on a gold electrode was chosen to determine neomycin. Polymyxin B sulphate and dexamethasone sodium phosphate do have a UV chromophore. So, a single LC method with UV detection was developed for the determination of polymyxin B sulphate and dexamethasone sodium phosphate. The sample pretreatment is simply done by diluting the formulation with water. For each method, the influence of the different chromatographic parameters on the separation, the interference of other active compounds and excipients, the repeatability and the linearity were investigated. Finally, the content of the actives in the formulation was studied at 0, 2, 4, 6, and 8 weeks. PMID- 15533668 TI - Development and validation of a liquid chromatographic method for determination of related-substances of mosapride citrate in bulk drugs and pharmaceuticals. AB - An isocratic reversed-phase high-performance liquid chromatographic (RP-HPLC) method for determination and evaluation of purity of mosapride citrate in bulk drugs and pharmaceuticals has been developed using Waters Symmetry C(18) column with acetonitrile:0.024 M orthophosphoric acid (28:72, v/v) adjusted to pH 3.0 with triethylamine and photodiode array detector set at 276 nm. The method is simple, rapid, selective and capable of detecting all process related impurities at trace levels in the finished products with detection limits ranging in between 0.2 x 10(-8)g and 6.4 x 10(-8)g. The method has been validated with respect to accuracy, precision, linearity, ruggedness, and limit of detection and quantification. The linearity range was 125-1000 microg/ml. The percentage recoveries from pharmaceutical dosages were ranged from 95.53 to 100.7. The method was found to be suitable not only for monitoring the reactions during the process development but also quality assurance of mosapride citrate. PMID- 15533669 TI - ICH guidance in practice: establishment of inherent stability of secnidazole and development of a validated stability-indicating high-performance liquid chromatographic assay method. AB - The degradation behaviour of secnidazole was investigated under different stress degradation (hydrolytic, oxidative, photolytic and thermal) conditions recommended by International Conference on Harmonisation (ICH) using HPLC and LC MS. A stability-indicating HPLC method was developed that could separate drug from degradation products formed under various conditions. Secnidazole was found to degrade significantly in alkaline conditions, oxidative stress, and also in the presence of light. Mild degradation of the drug occurred in acidic and neutral conditions. The drug was stable to dry heat. Resolution of drug and the degradation products formed under different stress studies were successfully achieved on a C-18 column utilizing water-methanol in the ratio of 85:15 and at the detection wavelength of 310 nm. The method was validated with respect to linearity, precision (including intermediate precision), accuracy, selectivity and specificity. PMID- 15533670 TI - Near infrared spectroscopy for qualitative comparison of pharmaceutical batches. AB - Pharmaceuticals are produced according to current pharmacopoeias, which require quality parameters. Tablets of identical formulation, produced by different factories should have the same properties before and after storage. In this article, we analyzed samples having two different origins before and after storage (30 degrees C, 75% relative moisture). The aim of the study is to propose two approaches to understand the differences between origins and the storage effect by near infrared spectroscopy. In the first part, the main wavelengths are identified in transmittance and reflectance near infrared spectra in order to identify the major differences between the samples. In this paper, this approach is called fingerprinting. In the second part, principal component analysis (PCA) is computed to confirm the fingerprinting interpretation. The two interpretations show the differences between batches: physical aspect and moisture content. The manufacturing process is responsible for the physical differences between batches. During the storage, changes are due to the increase of moisture content and the decrease of the active content. PMID- 15533671 TI - Development and validation of a LC-method for determination of catechin and epicatechin in aqueous extractives from leaves of Maytenus ilicifolia. AB - A reverse phase-LC method was developed and validated for separation and quantification of catechin and epicatechin in aqueous extractives from leaves of Maytenus ilicifolia. The analysis was performed using a C(18) column with acetic acid-acetonitrile gradient elution. The detection was carried out by UV at 280 nm and the peak identification was based on the retention times and by co chromatography with reference substances. High coefficients of determination were achieved for both catechin and epicatechin peaks from the standard solutions (0.9996 and 0.9999), as well as from extractives (0.9981 and 0.9982, respectively). The method showed good repeatability (R.S.D. <1.5%), reproducibility (R.S.D. <5%) and good accuracy for both catechin and epicatechin peaks (101.4%, R.S.D. = 1.18% and 100.6%, R.S.D. = 2.07%, respectively). PMID- 15533672 TI - Investigation of radiosterilization and dosimetric features of sulfacetamide sodium. AB - In the present work, the spectroscopic and kinetic features of the radical species induced in gamma-irradiated sulfacetamide-sodium (SS) was studied at room and at different temperatures in the dose range of 5-50kGy by electron spin resonance (ESR) technique with the aim of investigating the possibility of radiosterilization and dosimetric features of SS. A model consisting of four radical species of different spectroscopic features denoted as I-IV was found to describe best whole experimental data derived throughout the present work. These species were quite stable at room temperature but relatively unstable above room temperature. Heights of the characteristic resonance peaks measured with respect to the base line were considered to monitor microwave, temperature, time dependent and kinetic features of the radical species contributing to ESR spectrum. Collected experimental data were used to characterize the radical species responsible from ESR spectra through simulation calculations. Possible changes in the IR bands of gamma irradiated SS was also investigated by FT-IR technique, but no definite difference was observed between unirradiated and irradiated IR spectra of SS. As in other sulfonamides, radiation yield of solid SS was found to be very low (G 0.9913) and low root mean square errors of prediction (RMSEP < 0.094%). Usually the main water bands are "cut out" of the spectra to improve the model, however this is associated with the loss of relevant spectroscopic information. Thus, the entire spectrum including the OH bands is used, as these are not only found in water but also in the kavapyrones. The use of this new strategy succeeds in overcoming the difficulties in NIRS and establishes NIRS as a valid alternative in the routine quality control of plant extracts. PMID- 15533681 TI - Development of a rapid RP-HPLC method for the determination of clonazepam in human plasma. AB - A rapid and sensitive high-performance liquid chromatography method with UV detection was developed for the determination of clonazepam in human plasma using 3-methylclonazepam, as internal standard. A one-step extraction of both compounds was performed with a mixture of hexane/ethyl acetate (90:10, v/v). The HPLC analysis was carried out on a Nova Pak((R)) C(18) reversed-phase column with a mobile phase of acetonitrile-0.01 M sodium acetate adjusted to pH 7 with dilute acetic acid (40:60, v/v). A linear response was observed over the concentration range 5-100 ng/mL. Intra- and inter-day assay precision and accuracy fulfilled the international requirements. The lower limit of quantification was 5 ng/mL without interference of endogenous components. For analytical purpose, the stability of clonazepam in bidistilled water and plasma has been studied. A rapid degradation was noticed when clonazepam was stored in bidistilled water at the daylight following a first-order kinetic rate with a 87 min half life whereas no significant degradation was observed in plasma. This method was applied to measure plasma concentrations of clonazepam either in patients receiving therapeutic doses or in poisoning cases. PMID- 15533682 TI - A high-performance liquid chromatographic method for determination of praziquantel in plasma. AB - A simple, sensitive, selective and reproducible method based on a reversed-phase chromatography was developed for the determination of praziquantel in human plasma. Praziquantel was separated from the internal standard (diazepam) on a Luna C18 column (250 mm x 4.6mm, 5 microm particle size), with retention times of 4.8 and 6.2 min, respectively. Ultraviolet detection was set at 21 7 nm. The mobile phase consisted of acetonitrile and distilled water (70:30, v/v), running through the column at a flow rate of 1.0 ml/min. The chromatographic analysis was operated at 25 degrees C. Sample preparation (1 ml plasma) was done by a single step liquid-liquid extraction with the mixture of methyl-tert-butylether and dichloromethane at the ratio of 2:1 (v/v). Calibration curves in plasma at the concentrations 0, 50, 100, 200, 400, 800 and 1600 ng/ml were all linear with correlation coefficients better than 0.999. The precision of the method based on within-day repeatability and reproducibility (day-to-day variation) was below 15% (relative standard deviation: R.S.D.). Good accuracy was observed for both the intra-day or inter-day assays, as indicated by the minimal deviation of mean values found with measured samples from that of the theoretical values (below +/ 15%). Limit of quantification (LOQ) was accepted as 5 ng using 1 ml samples. The mean recovery for praziquantel and the internal standard were greater than 90% for both praziquantel and internal standard. The method was free from interference from the commonly used antibiotic and antiparasitic drugs. The method appears to be robust and has been applied to a pharmacokinetic study of praziquantel in three healthy Thai volunteers following a single oral dose of 40 mg/kg body weight praziquantel. PMID- 15533683 TI - Simple and rapid determination of adenosine in human synovial fluid with high performance liquid chromatography-mass spectrometry. AB - A simple, fast, sensitive and selective reversed-phase high performance liquid chromatography-mass spectrometry coupling with an electrospray ionization (ESI) interface method is described for the determination of adenosine in human synovial fluid. This method involved the use of the [M + H](+)ions of adenosine and 2-chloroadenosine (internal standard for the assay) at m/z 268 and 302 in positive ion mode with selective ion monitoring (SIM). Separation was carried out on a 2.0 x 150 mm Shimadzu VP-ODS column by using an isocratic elution with a mobile phase consisting of water (94%),methanol (5%) and formic acid (1%). No interference with the components of the biological matrix was observed in the determination conditions. The calibration curve was linear in the range of 0.2 140 microgml(-1). The limits of quantification (LOQ) and detection (LOD) were 0.2 and 0.03 microgml(-1), respectively. The standard recoveries were between 93.3 and 104.0%. The method was successfully applied to determination of adenosine in some synovial fluids of patients affected by rheumatoid arthritis. PMID- 15533684 TI - A liquid chromatographic-tandem mass spectrometric method for the determination of YM466, a novel Factor Xa inhibitor, in rat plasma. AB - A sensitive and selective method for the determination of YM466, a novel Factor Xa inhibitor, in rat plasma was developed and validated using liquid chromatography-mass spectrometry with electrospray ionization. Plasma samples were pretreated by solid-phase extraction and chromatographed on a C18 column (75 mm x 4.6 mm i.d.) with a mobile phase consisting of 0.1% formic acid-methanol (70:30 (v/v)), and detected using selected reaction monitoring in the positive ion mode. The lower limit of quantification was 0.4 ng/ml, and good precision and accuracy were achieved. The validated method allowed analysis of samples for the determination of pharmacokinetic profiles of YM466 in rats. PMID- 15533685 TI - On-line assay of the S-enantiomers of enalapril, ramipril and pentopril using a sequential injection analysis/amperometric biosensor system. AB - A sequential injection analysis/amperometric biosensor system is proposed for the enantioselective analysis of the S-enantiomer of enalapril, ramipril and pentopril. The amperometric biosensor used as detector in the sequential injection analysis was designed by immobilization of l-amino acid oxidase in carbon paste. The proposed SIA system can be utilized reliably for the enantioanalysis of the S-enantiomer from the raw materials as well as from their pharmaceutical formulations, with a rate of 75 samples per hour and R.S.D. values better than 0.1% (n = 10). PMID- 15533686 TI - A validated cold vapour-AAS method for determining mercury in human red blood cells. AB - A cold vapour-atomic absorption spectrometry (CV-AAS) method for determining mercury in dental students and clinical teaching staff red blood cells at a dental school using amalgam as a restorative material has been validated. A number of blood samples (n = 122) from dental students in years I to V, clinical teachers in restorative dentistry and controls were collected and analysed. Accuracy, linearity, precision (repeatability and reproducibility) and robustness of the method have been determined, and detection and quantification limits have been calculated. Linearity of response was verified for concentrations ranging from 5 to 40 microgL(-1) of mercury. Correlation coefficient of the calibration straight lines was always >/=0.99. Intra-day precision of the method gave coefficient of variation (CV) of 5.51%. Inter-day precision of the method calculated after analysis of five different concentrations of mercury standard solutions by the same analyst in different days and by two different analysts in different days gave coefficient of variation 4.89 and 5.44%, respectively. The accuracy of the method was calculated a CRM NIST 966 (toxic metals in bovine blood) total amount of mercury was found a concentration of 28.83 +/- 2.2 microgL(-1). Recovery was 89.27%. Robustness of the method evaluated by changing different experimental conditions under which analyses performed, fractional factorial design was done for assessing robustness of the method. Root mean square error was found out as 1.56. Limits of detection and quantification were 1.84 and 4.03 microg of Hg per litre of sample, respectively. Results show the suitability of the method for direct measurement of mercury in red blood cells and the importance of the working conditions for people dealing with amalgam at a dental school. PMID- 15533687 TI - Adsorptive properties of cefpodoxime proxetil as a tool for a new method of its determination in urine. AB - It was found that the reduction of the cefpodoxime proxetil (CP) molecule is strongly influenced by the adsorption. The adsorptive properties of CP were investigated in order to achieve an increase sensitivity of its determination. Validated adsorptive stripping differential pulse voltammetry is applied for the determination of low concentration of CP at pH 3.5 and 9.0 where the best pronounced adsorption effects were observed. The linearity of the calibration curves were achieved from 1 x 10(-8) to 1 x 10(-7)M with limit of detection (LOD) of 6.3 x 10(-9) and 7.1 x 10(-9)M, and limit of quantification (LOQ) of 2.1 x 10( 8) and 2.3 x 10(-8)M, at pH 3.5 and 9.0, respectively. The proposed method was tested for CP determination in spiked urine samples, enabling determination of low concentrations of CP. PMID- 15533688 TI - Overestimation of rifampicin during colorimetric analysis of anti-tuberculosis products containing isoniazid due to formation of isonicotinyl hydrazone. AB - When present together in fixed-dose combinations (FDC) of anti-tuberculosis drugs, rifampicin (R) and isoniazid (H) interact with each other to form isonicotinyl hydrazone (HYD). In a preliminary study, this product was found to possess similar colorimetric spectrum to that of rifampicin. Therefore, an investigation was undertaken to determine interference of HYD during analysis of rifampicin in FDC products by colorimetry. For the purpose, standard plots were constructed for rifampicin and HYD at 475 nm, the wavelength maximum for both the compounds. The plots were linear in the range of 10-100 microg/ml. Molar absorptivity values for rifampicin and HYD were 15279 and 5034, respectively. It indicated that HYD possessed one-third absorptivity to that of rifampicin. The analysis of combinations of rifampicin and HYD revealed that rifampicin could be overestimated to a maximum extent of 33%, while interference varied at other relative ratios of the two compounds. This was also confirmed by colorimetric and HPLC analysis of a degraded marketed product and samples from a dissolution study. Thus this investigation suggests that any method devoid of interference of HYD should be preferred for analysis of rifampicin, whenever it is present along with isoniazid. PMID- 15533689 TI - Determination of rosiglitazone in coated tablets by MEKC and HPLC methods. AB - Micellar electrokinetic chromatographic (MEKC) and high-performance liquid chromatographic (HPLC) methods were developed and subsequently validated for the determination of rosiglitazone (RSG) in coated tablet, a potent new oral antihyperglicemic agent. The electrophoretic separation was performed in a fused silica capillary of total length 48.0 cm (effective length 39.5 cm, 75 microm i.d.) using 10 mM sodium tetraborate buffer (pH 9.0) containing 30 mM sodium dodecyl sulfate (SDS) as the background electrolyte (BGE). The separating voltage used was of 20 kV at 25 degrees C and the diode array detector was set at 247 nm. The MEKC method was compared with HPLC method using a RP-18 column (125 x 4.0mm i.d.) eluted with a mobile phase consisting of mixture of 25 mM potassium dihydrogen phosphate buffer and acetonitrile (55:45, v/v), adjusting the pH to 6.2 with dilute potassium hydroxide. Statistical analysis by Student's t-test showed no significant differences between the results obtained by two methods. The results indicated that MEKC can be used an alternative method to HPLC for the determination of rosiglitazone in pharmaceutical dosage form. PMID- 15533690 TI - Study of the interaction between monoammonium glycyrrhizinate and bovine serum albumin. AB - The interaction between monoammonium glycyrrhizinate (MAG) and bovine serum albumin (BSA) were studied by fluorescence and absorption spectroscopy. The quenching mechanism of fluorescence of bovine serum albumin by monoammonium glycyrrhizinate was discussed. The binding sites number n and apparent binding constant K were measured by fluorescence quenching method. The thermodynamic parameters DeltaH degrees , DeltaG degrees , DeltaS degrees at different temperatures were calculated. The distance r between donor (bovine serum albumin) and acceptor (monoammonium glycyrrhizinate) was obtained according to Forster theory of non-radiation energy transfer. The results of synchronous fluorescence spectra and UV-vis absorption spectra show that the conformation of bovine serum albumin has been changed. PMID- 15533691 TI - MMSPE-RP-HPLC method for the simultaneous determination of methimazole and selected metabolites in fish homogenates. AB - A simple and rapid high-performance liquid chromatography method with ultraviolet detection was developed for the determination of the thyreostatic compound methimazole (1-methyl-2-mercaptoimidazole, MET) and metabolites 2 mercaptoimidazol (SHMET), N-methylthiourea (MTU) and N-methylhydantoin (MEH) from zebrafish (D. rerio) whole body homogenates using mixed mode solid phase extraction technique for sample pre-treatment. The highly polar compounds were separated on a difunctionally bonded silica based reverse phase column using gradient elution. Retention factors ranged between 1.53 and 5.66. The method was linear between 0.1 and 30 microg/ml, the detection limits were 0.4 ng for MET and SHMET, 0.6 ng for MTU and 2.6 ng for MEH. Extraction method was exhibited average recovery rates of 85.2-97.6%. PMID- 15533692 TI - Determination and in-process control of zolpidem synthesis by high-performance liquid chromatography. AB - A high-performance liquid chromatographic assay with diode-array detection has been developed for the in-process control of zolpidem synthesis and for the analysis of the drug and its synthetic intermediates. The separation uses a 4.6mm i.d. reversed-phase Kromasil C(18) (150 mm) column, 5 microm particle size with a gradient elution mode of acetonitrile and 0.02 M NH(4)OAc (adjusted to pH 8.0) as the mobile phase (flow rate 1.0 mlmin(-1)). The analysis is performed in 12 min. The method is simple, rapid and highly specific. PMID- 15533693 TI - Investigation of profile difference between Alzheimer's disease patients declining at different rates: examination of baseline neuropsychological data. AB - The rate of cognitive decline in AD has been noted to vary significantly among patients. The ability to predict the probable rate of decline early in the disease process would be of great practical importance. Attempts to analyze early cognitive deficits to find patterns associated with rapid decline have met with limited success. This paper utilized a large sample of patients with a diagnosis of probable AD evaluated longitudinally in ongoing research at the ADRC at Baylor College of Medicine and a statistical procedure of profile analysis to assess the initial data for a pattern associated with rapid decline. The findings indicated that despite initial equality of MMSE scores, patients showing rapid MMSE decline at one year displayed significantly more impaired performance on neuropsychological measures at diagnosis. Discussion includes discussion on the use of the MMSE for tracking general cognitive function and the difficulties of ascertaining stable profiles for prediction. PMID- 15533694 TI - Nonpatient CPT performance varying target frequency and interstimulus interval on five response measures. AB - Studies of sustained attention using the Continuous Performance Test (CPT) often incorporate effortful processing conditions and different response measures. Difficulty of attentional effort and interrelationships among response measures, however, have not been well established particularly with normal populations. This study investigated three CPT tasks which varied in interstimulus interval and target frequency in 107 nonpatient adults. This report also examined five response measures (d', a measure of discriminability; In(beta), a measure of response bias; reaction time; and omission and commission errors). Findings indicated that a high target frequency condition was more effortful than a short interstimulus interval condition on all response measures. Further, d', omission errors, and commission errors differentiated the short interstimulus interval condition from baseline, but reaction time and In(beta) did not. Participants with faster reaction times in the high target frequency condition had fewer omission errors but more commission errors; these relationships may have been mediated by response bias. This suggested that response measures were interrelated but nonredundant, and that these relationships were influenced by attentional effort and response bias. PMID- 15533695 TI - Wisconsin Card Sorting Test with children: a meta-analytic study of sensitivity and specificity. AB - More and more frequently the presence of executive function deficits appears in the research literature in conjunction with disabilities that affect children. Research has been most directed at the extent to which executive function deficits may be implicated in specific disorders such as attention deficit hyperactivity disorder (ADHD); however, deficits in executive function have been found to be typical of developmental disorders in general. The focus of this paper is to examine the extent to which one frequently used measure of executive function, the Wisconsin Card Sorting Test (WCST), demonstrates sensitivity and specificity for the identification of those executive function deficits associated with ADHD as well as its use with other developmental disorders through meta-analytic methods. Evidence of sensitivity of the WCST to dysfunction of the central nervous system is reviewed. Effect sizes calculated for all studies compared groups of children on differing variables of the WCST. The results of this meta-analysis suggest that across all of the studies, individuals with ADHD fairly consistently exhibit poorer performance as compared to individuals without clinical diagnoses on the WCST as measured by Percent Correct, Number of Categories, Total Errors, and Perseverative Errors. Notably, other various clinical groups performed more poorly than the ADHD groups in a number of studies. Thus, while impaired performance on the WCST may be indicative of an underlying neurological disorder, most likely related to frontal lobe function, poor performance is not sufficient for a diagnosis of ADHD. Implications for further research are presented. PMID- 15533696 TI - The operating characteristics of the major HRNES-R measures. AB - The operating characteristics and base rate effects of tests and indexes in the Halstead Russell Neuropsychological Evaluation System-Revised (HRNES-R) were obtained to determine its accuracy in assessing brain damage. Since operating characteristics and base rate problems are not well understood they were discussed in some detail. The operating characteristics of Sensitivity, Specificity, Positive Predictive Power, Negative Predictive Power and Overall Predictive Power along with base rate effects were obtained for 2 HRNES-R indexes, 10 index tests and 3 other Halstead Reitan Battery (HRB) tests. The indexes were found to be as accurate as the most accurate indexes from other HRB studies. The accuracy of the various tests were high but varied according to their function and design. PMID- 15533697 TI - The relationship between motor coordination, executive functioning and attention in school aged children. AB - Given the high level of comorbidity of attention deficit hyperactivity disorder (ADHD) and developmental coordination disorder (DCD), deficits in executive function (EF), shown to be present in children with ADHD, may also be implicated in the motor coordination deficits of children with DCD. The aim of this study was to explore the relationship between EF and motor ability. A sample of 238 children, 121 girls and 117 boys, aged between 6 and 15 years was recruited for this project. Motor ability was assessed using the McCarron Assessment of Neuromuscular Development (MAND), level of inattention using the Child Behavior Checklist (CBCL), and Verbal IQ (VIQ) was estimated using subtests of the WISC III. A reaction time task and three EF tasks measuring response inhibition, working memory and the ability to plan and respond to goal-directed tasks were administered. It was found that motor ability significantly accounted for variance in tasks measuring speed of performance, whereas inattention appeared to influence performance variability. Despite past evidence linking poor motor ability with inattention, there was little overlap in the processes that are affected in children with motor coordination or attention problems. PMID- 15533698 TI - Validation of a Developmental Neuropsychological Assessment (NEPSY) through comparison of neurological, scholastic concerns, and control groups. AB - Validity of the standard, 13 subtest A Developmental Neuropsychological Assessment (NEPSY) was investigated by comparing scores for 30 children with neurological conditions, 35 children with scholastic concerns, and 39 controls. Overall differences were found among the groups with and without controlling for IQ (Lambda = .60, Lambda = .001; Lambda = .70, P < .001). Four of five NEPSY domain scores differed among the three groups. Language and Sensorimotor domain score differences were found even when IQ was controlled, and group status accounted for substantial variance in these domain scores. Regarding specific tasks, Phonological Processing and Fingertip Tapping were among the subtests that varied the most between groups, especially when children with scholastic concerns were compared with controls. Findings offer preliminary support for the validity of several NEPSY indexes. PMID- 15533699 TI - Commentary--neuropsychological evaluation of patients with traumatic brain injury: polarization versus holistic integration. PMID- 15533700 TI - Assumptions that underlie predicting premorbid IQ: a comment on the "evaluation of the accuracy of two regression-based methods for estimating premorbid IQ". PMID- 15533701 TI - Domains of life satisfaction in social anxiety disorder: relation to symptoms and response to cognitive-behavioral therapy. AB - A general sense of satisfaction with life has been shown to be discriminable from symptom levels and disability in clinical populations. The current study focused on the utility of identifying domains of life satisfaction in social anxiety disorder and differential changes in these domains following cognitive-behavioral group therapy (CBGT). An exploratory principal axis factor analysis of the items of the Quality of Life Inventory in clients with a principal diagnosis of social anxiety disorder (N=138) yielded four domains of life satisfaction: (1) Achievement, (2) Social Functioning, (3) Personal Growth, and (4) Surroundings. Prior to treatment, clients reported dissatisfaction in the Achievement and Social Functioning domains. Further, levels of satisfaction in these domains were significantly related to severity of social anxiety and depressive symptoms. Finally, analyses of a subsample of clients completing 12-weeks of cognitive behavioral group therapy revealed significant improvements in the Achievement and Social Functioning factors. These findings provide further support for the assertion that social anxiety disorder has important implications for clients' quality of life and that CBGT can successfully impact several domains of satisfaction. PMID- 15533702 TI - The looming maladaptive style predicts shared variance in anxiety disorder symptoms: further support for a cognitive model of vulnerability to anxiety. AB - Looming vulnerability pertains to a distinct cognitive phenomenology characterized by mental representations of dynamically intensifying danger and rapidly rising risk as one projects the self into an anticipated future [J. Pers. Soc. Psychol. 79 (2000) 837]. While looming appraisals can be experienced as state elicitation, some individuals are hypothesized to develop an enduring cognitive pattern of cross-situational looming appraisals, the looming maladaptive style (LMS), which functions as a cognitive vulnerability to anxiety. In the present study, we examined the extent to which the LMS predicts common variance in numerous anxiety disorder symptoms, independent of the potentially confounding effects of current depressive symptoms. Specifically, we hypothesized that controlling for depressive symptoms, LMS would predict shared variance in a latent factor comprised of indicators of five anxiety disorder symptoms: obsessive-compulsive disorder, post-traumatic stress disorder, generalized anxiety disorder, social phobia, and specific phobic fears. Measures of these anxiety disorder symptoms, depressive symptoms, and looming vulnerability were administered to unselected college student population. Structural equations modeling analyses provided support for our hypothesis that LMS predicts shared variance in anxiety disorder symptoms and suggest that this cognitive style may be an overarching dimension of vulnerability to anxiety. PMID- 15533703 TI - Covariation bias and its physiological correlates in panic disorder patients. AB - A covariation bias, i.e., the overestimation of random contingencies between fear relevant stimuli and aversive consequences, seems to characterize anxiety disorders. Panic patients (n=30) and healthy controls (n=25) were exposed to panic-relevant, neutral, and phobia-relevant but panic-irrelevant picture stimuli, followed randomly be aversive consequences (acoustic startle stimuli). While covariation estimates reflected objective contingencies in both groups, only panic patients revealed a more negative Contingent Negative Variation (CNV) to panic-relevant than to phobia-relevant and neutral pictures. For startle reflex, only main effects of picture category were found, indicating that valence effects of picture stimuli were not specifically distorted in panic patients. CNV presumably reflects a biased processing of disorder-relevant stimuli by panic patients, perhaps with the expectation that aversive consequences will follow these stimuli. PMID- 15533704 TI - Making sense of informant disagreement for overanxious disorder. AB - A community sample of 2798 8-17-year-old twins and their parents completed a personal interview about the child's current psychiatric history on two occasions separated by an average of 18 months. Parents also completed a personal interview about their own lifetime psychiatric history at entry to the study. Results indicate that informant agreement for overanxious disorder (OAD) was no better than chance, and most cases of OAD were based on only one informant's ratings. Disagreement about level of OAD symptoms or presence of another disorder (mostly phobias or depression) accounted for most cases of informant disagreement: 60% of cases based only on child interview, 67% of cases based only on maternal interview, and 100% of cases based only on paternal interview. OAD diagnosed only by maternal interview was also distinguished by an association with maternal alcoholism and increasingly discrepant parental reports of marital difficulties. Given the substantial overlap in case assignments for DSM-III-R OAD and DSM-IV GAD, these findings may identify sources of informant disagreement that generalize to juvenile GAD. PMID- 15533705 TI - Marital predictors of symptom severity in panic disorder with agoraphobia. AB - Twenty-six to forty percent of individuals suffering from panic disorder with agoraphobia (PDA) do not benefit significantly from cognitive-behavior therapy. Marital problems are among risk factors that may explain this limited impact. Some studies suggest that PDA treatment outcome is related to the couple's ability to communicate and solve problems during and after treatment. It may be also useful to further clarify the interplay of marital interpersonal variables with PDA severity before any intervention. This study aims at specifying the links between PDA symptom severity on the one hand and, on the other hand, marital adjustment, attachment style and personal problem-solving skills in both spouses. Results obtained from a group of 67 PDA patients (44 women and 23 men) and their partners showed that some PDA symptoms or comorbid depressive symptoms were more severe when both spouses independently scored low on problem-solving skills or marital adjustment, and when attachment style of PDA patients was insecure. Marital adjustment and difficulties in problem-solving, more specifically, avoidance of problem-solving activities in PDA patients, were the best predictors of PDA symptom severity. In light of these findings, a more complete program of problem-solving and acceptance strategies could be developed as part of a cognitive-behavior treatment of PDA. Other theoretical and clinical implications are discussed. PMID- 15533707 TI - Linkage disequilibrium maps constructed with common SNPs are useful for first pass disease association screens. AB - To develop an efficient strategy for mapping genetic factors associated with common diseases, we constructed linkage disequilibrium (LD) maps of human chromosomes 5, 7, 17, and X. These maps consist of common single nucleotide polymorphisms at an average intermarker distance of 100 kb. The genotype data from these markers in a panel of American samples of European descent were analyzed to produce blocks of markers in strong pair-wise LD. Power calculations were used to guide block definitions and predicted that high-level LD maps would be useful in initial genome scans for susceptibility alleles in case-control association studies of complex diseases. As anticipated, LD blocks on the X chromosome were larger and covered more of the chromosome than those found on the autosomes. PMID- 15533708 TI - Construction of representative transcript and protein sets of human, mouse, and rat as a platform for their transcriptome and proteome analysis. AB - The number of mammalian transcripts identified by full-length cDNA projects and genome sequencing projects is increasing remarkably. Clustering them into a strictly nonredundant and comprehensive set provides a platform for functional analysis of the transcriptome and proteome, but the quality of the clustering and predictive usefulness have previously required manual curation to identify truncated transcripts and inappropriate clustering of closely related sequences. A Representative Transcript and Protein Sets (RTPS) pipeline was previously designed to identify the nonredundant and comprehensive set of mouse transcripts based on clustering of a large mouse full-length cDNA set (FANTOM2). Here we propose an alternative method that is more robust, requires less manual curation, and is applicable to other organisms in addition to mouse. RTPSs of human, mouse, and rat have been produced by this method and used for validation. Their comprehensiveness and quality are discussed by comparison with other clustering approaches. The RTPSs are available at . PMID- 15533706 TI - A brief sleep scale for Posttraumatic Stress Disorder: Pittsburgh Sleep Quality Index Addendum for PTSD. AB - Sleep disturbances reflect a core dysfunction underlying Posttraumatic Stress Disorder (PTSD). Specifically, disruptive nocturnal behaviors (DNB) may represent PTSD-specific sleep disturbances. The Pittsburgh Sleep Quality Index Addendum for PTSD (PSQI-A) is self-report instrument designed to assess the frequency of seven DNB. The goal of this study was to examine the psychometric properties of the PSQI-A to characterize DNB in a group of participants with and without PTSD. Results indicate that the PSQI-A has satisfactory internal consistency and good convergent validity with two standard PTSD measures even when excluding their sleep-related items. A global PSQI score of 4 yielded a sensitivity of 94%, a specificity of 82%, and a positive predictive value of 93% for discriminating participants with PTSD from those without PTSD. The PSQI-A is a valid instrument for PTSD applicable to both clinical and research settings. PMID- 15533709 TI - Learnability-based further prediction of gene functions in Gene Ontology. AB - Currently the functional annotations of many genes are not specific enough, limiting their further application in biology and medicine. It is necessary to push the gene functional annotations deeper in Gene Ontology (GO), or to predict further annotated genes with more specific GO terms. A framework of learnability based further prediction of gene functions in GO is proposed in this paper. Local classifiers are constructed in local classification spaces rooted at qualified parent nodes in GO, and their classification performances are evaluated with the averaged Tanimoto index (ATI). Classification spaces with higher ATIs are selected out, and genes annotated only to the parent classes are predicted to child classes. Through learnability-based further predicting, the functional annotations of annotated genes are made more specific. Experiments on the fibroblast serum response dataset reported further functional predictions for several human genes and also gave interesting clues to the varied learnability between classes of different GO ontologies, different levels, and different numbers of child classes. PMID- 15533710 TI - Exploring the characteristics of sequence elements in proximal promoters of human genes. AB - Central to reconstruction of cis-regulatory networks is identification and classification of naturally occurring transcription factor-binding sites according to the genes that they control. We have examined salient characteristics of 9-mers that occur in various orders and combinations in the proximal promoters of human genes. In evaluations of a dataset derived with respect to experimentally defined transcription initiation sites, in some cases we observed a clear correspondence of highly ranked 9-mers with protein-binding sites in genomic DNA. Evaluations of the larger dataset, derived with respect to the 5' end of human ESTs, revealed that a subset of the highly ranked 9-mers corresponded to sites for several known transcription factor families (including CREB, ETS, EGR-1, SP1, KLF, MAZ, HIF-1, and STATs) that play important roles in the regulation of vertebrate genes. We identified several highly ranked CpG containing 9-mers, defining sites for interactions with the CREB and ETS families of proteins, and identified potential target genes for these proteins. The results of the studies imply that the CpG-containing transcription factor-binding sites regulate the expression of genes with important roles in pathways leading to cell-type-specific gene expression and pathways controlled by the complex networks of signaling systems. PMID- 15533711 TI - Genome physical mapping with large-insert bacterial clones by fingerprint analysis: methodologies, source clone genome coverage, and contig map quality. AB - Genome physical mapping with large-insert clones by fingerprint analysis is becoming an active area of genomics research. Here, we report two new capillary electrophoresis-based fingerprinting methods for genome physical mapping and the effects of different fingerprinting methods and source clone genome coverage on quality physical map construction revealed by computer simulations and laboratory experiments. It was shown that the manual sequencing gel-based two-enzyme fingerprinting method consistently generated larger and more accurate contigs, followed by the new capillary electrophoresis-based three-enzyme method, the new capillary electrophoresis-based five-enzyme (SNaPshot) method, the agarose gel based one-enzyme method, and the automatic sequencing gel-based four-enzyme method, in descending order, when 1% or fewer questionable clones were allowed. Analysis of clones equivalent to 5x, 8x, 10x, and 15x genomes using the fingerprinting methods revealed that as the number of clones increased from 5x to 10x, the contig length rapidly increased for all methods. However, when the number of clones was increased from 10x to 15x coverage, the contig length at best increased at a lower rate or even decreased. The results will provide useful knowledge and strategies for effective construction of quality genome physical maps for advanced genomics research. PMID- 15533712 TI - Timing of establishment of paternal methylation imprints in the mouse. AB - Imprinted genes are characterized by predominant expression from one parental allele and differential DNA methylation. Few imprinted genes have been found to acquire a methylation mark in the male germ line, however, and only one of these, H19, has been studied in detail. We examined methylation of the Rasgrf1 and Gtl2 differentially methylated regions (DMR) to determine whether methylation is erased in male germ cells at e12.5 and when the paternal allele acquires methylation. We also compared their methylation dynamics with those of H19 and the maternally methylated gene Snrpn. Our results show that methylation is erased on Rasgrf1, H19, and Snrpn at e12.5, but that Gtl2 retains substantial methylation at this stage. Erasure of methylation marks on Gtl2 appears to occur later in female germ cells to give the unmethylated profile seen in mature MII oocytes. In the male germ line, de novo methylation of Rasgrf1, Gtl2, and H19 occurs in parallel between e12.5 and e17.5, but the DMR are not completely methylated until the mature sperm stage, suggesting a methylation dynamic different from that of IAP, L1, and minor satellite sequences, which have been shown to become fully methylated by e17.5 in male germ cells. This study also indicates important differences between different imprinted DMR in timing and extent of methylation in the germ cells. PMID- 15533713 TI - Methylation at mouse Cdkn1c is acquired during postimplantation development and functions to maintain imprinted expression. AB - Monoallelic expression of imprinted genes is generally associated with differential methylation. Methylation may be inherited as the gametic imprinting mark or may be acquired postfertilization. Here, we characterize a differentially methylated region associated with the mouse Cdkn1c gene and find that it is confined to a CpG island that begins 600 bp 5' of the promoter and extends into the transcription unit. Our analysis indicates that methylation of this region is not inherited from sperm, is acquired specifically on the paternal allele following implantation, and is dependent on KvDMR1. We further demonstrate that although methylation is required for maintaining silencing of the paternal Cdkn1c allele, it is not a prerequisite for the establishment of monoallelic expression at this locus. Prior to the onset of differential methylation, additional epigenetic modifications must play a role in distinguishing the parental alleles of Cdkn1c and influencing their expression. PMID- 15533714 TI - Analysis of mouse germ-cell transcriptome at different stages of spermatogenesis by SAGE: biological significance. AB - The transcriptomes of mouse type A spermatogonia (Spga), pachytene spermatocytes (Spcy), and round spermatids (Sptd) were determined by sequencing the respective SAGE (Serial Analysis of Gene Expression) libraries. A total of 444,015 tags derived from one Spga, two Spcy, and one Sptd library were analyzed, and 34,619 different species of transcripts were identified, 5279 of which were novel. Results indicated the germ-cell transcriptome comprises of more than 30,000 transcripts. Virtual subtraction showed that cell-specific transcripts constitute 12-19.5% of the transcriptome. Components of the protein biosynthetic machinery are highly expressed in Spga. In Spcy transcription factors are abundantly expressed while transcripts encoding proteins involved in chromosome remodeling and testis-specific transcripts are prominent in Sptd. The databases generated by this work provide very useful resources for cellular localization of genes in silico. They are also extremely useful as sources for identification of splice variants of genes in germ cells. PMID- 15533715 TI - Expression analyses of human endogenous retroviruses (HERVs): tissue-specific and developmental stage-dependent expression of HERVs. AB - The evolutional and biological roles of human endogenous retroviruses (HERVs) are less recognized compared to those of L1. In the present study, we focused on the transcriptional activity of HERVs in normal human tissues and found five HERV loci that are actively expressed in normal tissues. All but one showed tissue specificity of expression: one was expressed in stomach and small intestine and three were in placenta. We subsequently examined by TaqMan-based RT-PCR assays the temporal expression profiles of the three placenta-specific HERVs along with syncytin and syncytin 2 and observed three patterns. Syncytin and HERV-Fb showed almost constant expression through gestations. Syncytin 2 gradually decreased as pregnancy proceeded. In contrast, expression from the HERV-H/F and HERV-K(HML-6) loci increased remarkably in term placentas. Term placentas in general showed larger interindividual differences in HERV expression levels. Our results suggest that HERVs might have more diverse effects than currently thought. PMID- 15533716 TI - Analysis of multiple Invs transcripts in mouse and MDCK cells. AB - Infantile nephronophthisis is associated with cystic kidneys, situs inversus, and INVS mutations. The function of the INVS product, inversin, is unknown but evidence suggests there are multiple inversin isoforms with differing molecular weights, cellular localization patterns, and binding partners. We used Northern blots, RT-PCR, and sequence analysis to identify alternative INVS transcripts. Northern blots probed with Invs cDNA detected four bands in normal mouse kidney. RT-PCR of mouse kidney RNA revealed Invs transcripts with skipping of exon 5, 11, or 13. We sequenced canine (MDCK-II cells) INVS and determined that the corresponding full-length protein shares identity with mouse (74%) and human (84%) inversin. Canine INVS produces a transcript that skips exon 12. Exon skips cause loss of inversin protein motifs, including ankyrin repeats, IQ domains, destruction boxes, and nuclear localization signals. Identification of INVS splice variants will help us determine which inversin protein motifs contribute to left-right asymmetry and kidney development. PMID- 15533717 TI - Patterns of coordinate down-regulation of ARE-containing transcripts following immune cell activation. AB - We evaluated the expression of over 900 AU-rich element (ARE)-containing transcripts in primary human T lymphocytes following stimulation with anti-CD3 and anti-CD28 antibodies and found that approximately 48% of these transcripts were regulated following T cell activation. We identified approximately 145 ARE containing transcripts that were rapidly induced and then rapidly disappeared within 1 h after activation. Another 250 ARE-containing transcripts expressed in resting T cells were rapidly turned off within 30 min after activation. The rates of transcript disappearance correlated well with rapid mRNA decay measured following transcriptional arrest with actinomycin D. We identified a subset of ARE-containing transcripts that were rapidly induced following T cell activation that were also induced following lipopolysaccharide stimulation of THP-1 monocytes, and these transcripts exhibited rapid decay in both cell types. Our results suggest that ARE-mediated mRNA decay plays an important role in the precisely coordinated down-regulation of gene expression following immune cell activation. PMID- 15533718 TI - Genes of glycolysis are ubiquitously overexpressed in 24 cancer classes. AB - Using NIH's public database dbEST for expression of genes and ESTs, genes of the glycolysis pathway have been found to be overexpressed in a set of 24 cancers representing more than 70% of human cancer cases worldwide. Genes can be classified as those that are almost ubiquitously overexpressed, particularly glyceraldehyde-3-phosphate dehydrogenase, enolase 1, and also pyruvate kinase, and those that are overexpressed in less than 50% of the investigated cancers. Cancers can be classified as those with overexpression of the majority of the glycolysis genes, particularly lymph node, prostate, and brain cancer, in which essentially all glycolysis genes are overexpressed, and those with only sporadic overexpression, particularly cancers of the cartilage or bone marrow. This classification may be useful when cancer therapies aimed at the Warburg effect are designed. PMID- 15533719 TI - IGF2 antisense transcript expression in porcine postnatal muscle is affected by a quantitative trait nucleotide in intron 3. AB - A paternally expressed quantitative trait locus (QTL) for muscle mass was mapped to the insulin-like growth factor II (IGF2) locus at the distal end of pig chromosome 2p. It was recently demonstrated that a G to A transition at position intron 3-nt 3072 is the quantitative trait nucleotide (QTN) underlying this QTL. In this study we report for the first time the existence of imprinted porcine IGF2 antisense transcripts (IGF2-AS) and demonstrate that their expression in postnatal muscle is also affected by the QTN. A coregulated expression of IGF2 and IGF2-AS RNAs in muscle and liver with decreasing transcription from fetal to adult age was demonstrated. Further, the significant difference found in expression of IGF2-AS in postnatal muscle between individuals with different QTL genotypes and the lack of significant differences in fetal muscle and liver reflect completely what has been found for IGF2 sense transcripts. PMID- 15533720 TI - Functional analysis of the murine Emr1 promoter identifies a novel purine-rich regulatory motif required for high-level gene expression in macrophages. AB - This study has investigated the transcriptional regulation of the Emr1 gene in murine macrophages and defined an enhancer element within the proximal promoter that is necessary for Emr1 expression in myeloid cells. This element consists of an extended purine-rich sequence (PuRS) of 83 consecutive purine residues containing 9 GGAA sequences, the core binding sequence for members of the Ets family of transcription factors. The Ets factor PU.1 associates with this PuRS element both in vitro and in vivo. Using a standard BLAST search we identified similar PuRS elements in other myeloid and nonmyeloid genes. All PuRS elements tested confer enhancer activity onto a heterologous promoter and chromatin immunoprecipitation experiments revealed that PU.1 associates in vivo with the PuRS elements from the genes expressed in myeloid cells. Our results provide evidence that extended purine-rich sequence elements may constitute a new transcription regulatory motif and that PU.1 association is a prerequisite for macrophage-specific expression. PMID- 15533721 TI - Mitochondrial DNA 3644T-->C mutation associated with bipolar disorder. AB - Mitochondrial dysfunction associated with mutant mitochondrial DNA (mtDNA) has been suggested in bipolar disorder, and comorbidity with neurodegenerative diseases was often noted. We examined the entire sequence of mtDNA in six subjects with bipolar disorder having comorbid somatic symptoms suggestive of mitochondrial disorders and found several uncharacterized homoplasmic nonsynonymous nucleotide substitutions of mtDNA. Of these, 3644C was found in 5 of 199 patients with bipolar disorder but in none of 258 controls (p = 0.015). The association was significant in the extended samples [bipolar disorder, 9/630 (1.43%); controls, 1/734 (0.14%); p = 0.007]. On the other hand, only 5 of 25 family members with this mutation developed bipolar disorder, of which 4 patients with 3644C had comorbid physical symptoms. The 3644T-->C mutation converts amino acid 113, valine, to alanine in the NADH-ubiquinone dehydrogenase subunit I, a subunit of complex I, and 113 valine is well conserved from Drosophila to 61 mammalian species. Using transmitochondrial cybrids, 3644T-->C was shown to decrease mitochondrial membrane potential and complex I activity compared with haplogroup-matched controls. According to human mitochondrial genome polymorphism databases, 3644C was not found in centenarians but was found in 3% of patients with Alzheimer disease and 2% with Parkinson disease. The result of modest functional impairment caused by 3644T-->C suggests that this mutation could increase the risk for bipolar disorder. PMID- 15533722 TI - A novel transmembrane MSP-containing protein that plays a role in right ventricle development. AB - We have identified and characterized a gene, Mospd3 on mouse chromosome 5 using gene trapping in ES cells. MOSPD3 is part of a family of proteins, including MOSPD1, which is defined by the presence of a major sperm protein (MSP) domain and two transmembrane domains. Interestingly Mospd3 is mammalian specific and highly conserved between mouse and man. Insertion of the gene trap vector at the Mospd3 locus is mutagenic and breeding to homozygosity results in a characteristic right ventricle defect and neonatal lethality in 50% of mice. The phenotypic defect is dependent on the genetic background, indicating the presence of genetic modifier loci. We speculate that the further characterization of Mospd3 will shed light on the complex genetic interactions involved in cardiac development and disease. PMID- 15533723 TI - X-linked hypoparathyroidism region on Xq27 is evolutionarily conserved with regions on 3q26 and 13q34 and contains a novel P-type ATPase. AB - X-linked hypoparathyroidism (HPT) has been mapped to a 988-kb region on chromosome Xq27 that contains three genes, MCF2/DBL, SOX3, and U7snRNA homologue, and a partial cDNA, AS6. We isolated the full-length AS6 cDNA, determined its genomic organization, and sought for abnormalities in HPT patients. AS6 was identified as the 3' UTR of ATP11C, a novel member of the P-type ATPases, which consists of 31 exons with alternative transcripts. The colocalization of ATP11C with SOX3 and MCF2/DBL on Xq27 mirrors that of ATP11A with SOX1 and MCF2L on 13q34 and ATP11B with SOX2 on 3q26. These colocalizations are evolutionarily conserved in mouse, and analyses indicate that SOX2 divergence likely occurred before the separation of SOX1 and SOX3. Analyses of ATP11C, MCF2, SOX3, and U7snRNA in HPT patients did not reveal mutations, implicating regulatory changes or mutation of an as yet unidentified gene in the etiology of X-linked hypoparathyroidism. PMID- 15533725 TI - Prognostic value of home heart rate for cardiovascular mortality in the general population: the Ohasama study. AB - BACKGROUND: Recently, the advantages of self-measurement of blood pressure (BP) at home have been recognized. The same advantages could also be applicable to resting heart rate (HR) values assessed at home using a device designed for home BP measurement. However, there have been no studies investigating whether home HR values predict the risk of cardiovascular disease mortality. We therefore investigated the usefulness of HR values in predicting cardiovascular mortality using a device that allowed self-measurement of BP and HR at home. METHODS: The association between the home-measured resting HR and the 10-year risk of cardiovascular mortality was examined in 1780 Japanese individuals >/=40 years of age who had no significant arrhythmias. A Cox proportional hazards model that adjusted for major risk factors was used. RESULTS: An increase of 5 beats/min in the morning home HR measurement was associated with a 17% increase in the risk of cardiovascular mortality (95% confidence interval 5% to 30%). This relationship was also statistically significant after adjustment for home BP values. Even when home-measured systolic BP was within the normal range (<135 mm Hg), subjects with HR >/=70 beats/min had a higher risk of cardiovascular mortality (relative hazard 2.16, 95% confidence interval 1.21 to 3.85) than those with normal systolic BP and HR values. CONCLUSIONS: Self-measurement of HR at home, together with self measurement of BP, is a simple method of providing useful clinical information for assessing cardiovascular risk. PMID- 15533724 TI - Structural evolution of the BRCA1 genomic region in primates. AB - Segmental duplications account for up to 6% of the human genome, and the resulting low-copy repeats (LCRs) are known to be associated with more than 20 genomic disorders. Many such duplication events coincided with the burgeoning of the Alu repeat family during the last 50 million years of primate evolution, and it has been suggested that the two phenomena might be causally related. In tracing the evolution of the BRCA1 17q21 region through the primate clade, we discovered the occurrence over the last 40 million years of a complex set of about eight large gene-conversion-mediated rearrangements in the approximately 4 Mb surrounding the BRCA1 gene. These have resulted in the presence of large and probably recombinogenic LCRs across the region, the creation of the NBR2 gene, the duplication of the BRCA1/NBR1 promoter, the bisection of the highly conserved ARF2 gene, and multiple copies of the KIAA0563 gene. The junctions lie within AluS repeats, members of an Alu subfamily which experienced massive expansion during the time that the rearrangements occurred. We present a detailed history of this region over a critical 40 million-year period of genomic upheaval, including circumstantial evidence for a causal link between Alu family expansion and the rearrangement-mediated destruction and creation of transcription units. PMID- 15533726 TI - Relationship between insulin resistance and end-organ damage in white coat hypertension. AB - BACKGROUND: End-organ damage is seen in some patients suffering from white coat hypertension (WCH). It remains unclear which patients elude the risk of end-organ damage. A relationship between end-organ damage and insulin resistance was hypothesized. METHODS: This hypothesis was tested by comparing two groups of patients with WCH: those presenting with end-organ damage and those without end organ damage. Both groups were tested for the presence of insulin resistance. The study was conducted in the outpatient clinics of the Internal and Family Medicine Departments of Cerrahpasa Medical School, Istanbul University. In this study 290 patients were screened for WCH, of which 75 were included in the study. The WCH was defined as an office blood pressure (BP) >/=140/90 mm Hg and mean daytime BP <135/85 mm Hg. The patients were divided into two groups according to the presence of end-organ damage. All patients received 24-h ambulatory BP monitoring. End-organ damage was diagnosed by revealing left ventricular hypertrophy on echocardiographic examination or retinopathy with fundoscopic examination. The homeostasis model assessment insulin resistance index (HOMA-IR) was used to assess insulin resistance. RESULTS: Among 75 patients with WCH, hypertensive retinopathy was detected in 25 patients, left ventricular hypertrophy in 15 patients, and both retinopathy and left ventricular hypertrophy simultanously in 6 patients. There were no significant differences between the groups regarding age, sex, lipid profile, and glucose levels. The HOMA-IR values in the patients with WCH and end-organ damage (4.2 +/- 1.7) were significantly higher (P < .0001) than those determined in patients with WCH but without end organ damage (2.6 +/- 1.8). In patients with WCH with HOMA-IR values above 3 the risk of end-organ damage was found to be higher. CONCLUSIONS: A possible relationship between end-organ damage and insulin resistance in patients with WCH exists. PMID- 15533727 TI - Self-measured home blood pressure in predicting ambulatory hypertension. AB - BACKGROUND: Physicians are commonly uncertain whether a person with office blood pressure (BP) around 140/90 mm Hg actually has hypertension. This is primarily because of BP variability. One approach is to perform self-measured home BP and determine if home BP is elevated. There is a general agreement that if home BP is >/=135/85 mm Hg, then antihypertensive therapy may be commenced. However, some persons with home BP below this cut-off will have ambulatory hypertension. We therefore prospectively study the role of home BP in predicting ambulatory hypertension in persons with stage 1 and borderline hypertension. METHODS: We studied in a cross-sectional way home and ambulatory BP in a group of 48 patients with at least two elevated office BP readings. The group was free of antihypertensive drug therapy for at least 4 weeks and performed 7 days of standardized self-BP measurements at home. We examined the relationships of the three BP methods and also defined a threshold (using receiver operating curves) for home BP that captures 80% of ambulatory hypertensives (awake BP >/=135/85 mm Hg). RESULTS: Office systolic BP (145 +/- 13 mm Hg) was significantly higher than awake (139 +/- 12 mm Hg, P = .013) and home (132 +/- 11 mm Hg, P < .001) BP. Office diastolic BP (88 +/- 4 mm Hg) was higher than home diastolic BP (80 +/- 8 mm Hg, P < .001) but not different from awake diastolic BP (88 +/- 8 mm Hg, P = .10). Home BP had a higher correlation (compared with office BP) with ambulatory BP. The home BP-based white coat effect correlated with ambulatory BP-based white coat effect (r = 0.83, P = .001 for systolic BP; r = 0.68, P = .001 for diastolic BP). The threshold for home BP of 80% sensitivity in capturing ambulatory hypertension was 125/76 mm Hg. CONCLUSIONS: Our preliminary data suggest that a lower self-monitored home BP threshold should be used (to exclude ambulatory hypertension) in patients with borderline office hypertension. PMID- 15533728 TI - Relationship of body composition to stress-induced pressure natriuresis in youth. AB - BACKGROUND: The contribution of stress to obesity-related cardiovascular disease is uncertain. The purpose of this study was to examine the influence of body composition on stress-induced pressure natriuresis. METHODS: Dual energy x-ray absorptiometry was performed in 127 African American and white youths to assess lean body mass (LBM), fat mass (FM), and total percentage of body fat (%BF). The stress protocol was comprised of a 2-h baseline period, 1-h video game competition stressor, and 2-h recovery period. Blood and urine samples were collected hourly and blood pressure (BP) was obtained at 15-min intervals. RESULTS: Both BP and urinary sodium excretion(U(Na)V) increased from baseline to stress and returned to prestress levels after stress (P = .001 for each). The BP levels and changes were positively correlated with LBM. In contrast, levels and changes in sodium excretion U(Na)V were inversely correlated with FM and %BM. Multiple regression analyses that included ethnicity, sex, angiotensin II (Ang II), and measures of body composition in the models indicated the following: a) LBM was the best predictor of stress systolic BP and independently contributed with ethnicity to stress diastolic BP; b) ethnicity was the only independent predictor of the stress-related change in systolic and diastolic BP; c) LBM was the only independent predictor of the change in BP from stress to recovery for both systolic and diastolic BP; and d) total percent body fat accounted for 11.2% of the variance of stress U(Na)V, with Ang II contributing an additional 6.1%. CONCLUSIONS: Based on the results of this study, ethnicity and body composition are related to stress-induced pressure natriuresis. PMID- 15533729 TI - Characteristics of hypertension in young adults with autosomal dominant polycystic kidney disease compared with the general U.S. population. AB - BACKGROUND: Patients with autosomal dominant polycystic kidney disease (ADPKD) often develop hypertension before any abnormalities in renal function are detected clinically. Therefore, standard screening (serum creatinine and urinalysis) of young individuals with unexplained hypertension to exclude renal parenchymal disease would rarely detect ADPKD. METHODS: Data from 516 subjects with ADPKD (217 male and 299 female), aged newbornto 55 years with a normal serum creatinine and no proteinuria based on urine dipstick, studied between 1985 and 2000, were compared with data from similar subjects from the National Health and Nutrition Examination Survey (NHANES) III (1988-1994) and NHANES IV (1999-2000) data, by gender. RESULTS: There was a highly significant occurrence of hypertension in young patients with ADPKD when compared to patients aged 20 to 34 years in the U.S. population. The hypertension in patients with ADPKD occurred in the absence of abnormal renal function or abnormal urinalysis. CONCLUSIONS: These data indicate that renal ultrasound screening of young hypertensive individuals (aged 20 to 34 years) should be considered when searching for causes of secondary hypertension. Identifying affected ADPKD individuals early in their disease will permit aggressive blood pressure treatment and early inhibition of the renin angiotensin-aldosterone system, which has been shown to reverse left ventricular hypertrophy, an important cardiovascular risk factor. In the present era of renal replacement therapy, cardiovascular complications are the main cause of death in patients with ADPKD. PMID- 15533730 TI - Prognostic value of left ventricular concentric remodeling in uncomplicated mild hypertension. AB - BACKGROUND: The prognostic value of left ventricular (LV) concentric remodeling in essential hypertension, and particularly in uncomplicated mild hypertension, is not yet completely clear. We investigated cardiovascular outcome in uncomplicated mild hypertensive patients with normal LV geometry and LV concentric remodeling. METHODS: The occurrence of fatal and nonfatal cardiovascular events was evaluated in 1088 uncomplicated mild hypertensive patients, 751 with normal LV geometry (LV mass index <125 g/m(2) in men and <110 g/m(2) in women and relative wall thickness <0.45) and 337 with LV concentric remodeling (LV mass index <125 g/m(2) in men and <110 g/m(2) in women and relative wall thickness >/=0.45). RESULTS: During follow-up (4.74 +/- 2.4 years, range 0.5 to 9.7 years) the event rates per 100 patient-years in subjects with normal LV geometry and LV concentric remodeling were 0.69 and 1.87, respectively. After adjustment for other covariates, Cox regression analysis showed that LV concentric remodeling (LV concentric remodeling versus normal LV geometry, RR 1.78, 95% CI 1.02 to 3.1, P < .05) was an independent predictor of cardiovascular events. CONCLUSION: In a population with uncomplicated mild hypertension, patients with LV concentric remodeling have a worse prognosis than those with normal LV geometry. PMID- 15533731 TI - Type 5 phosphodiesterase inhibition by sildenafil abrogates acute smoking-induced endothelial dysfunction. AB - BACKGROUND: Endothelial dysfunction is a key early event in the process of atherosclerosis and a risk factor for cardiovascular events. Sildenafil, an effective oral treatment for patients with erectile dysfunction, inhibits cGMP degradation by specific type 5 phosphodiesterase (PDE) inhibition. Sildenafil has been shown to improve vascular function, however, the effect of type 5 PDE inhibition on acute smoking-induced endothelial dysfunction is unknown. METHODS: We studied the effect of 50 mg of sildenafil on acute smoking-induced endothelial dysfunction in 14 male smokers according to a randomized, placebo-controlled, cross-over design. Endothelial function was evaluated with flow-mediated dilatation (FMD) of the brachial artery using high-resolution ultrasonography. RESULTS: Sildenafil abolishes the decrease in FMD of the brachial artery that is induced acutely by smoking (placebo/smoking session: from 4.56% +/- 0.60% to 2.80% +/- 0.43%, sildenafil/smoking session: from 3.83% +/- 0.64% to 4.33% +/- 0.47%, ie, improvement of 51%, P < .05). This was associated with no reversal effect of sildenafil on smoking-induced decrease in resting brachial artery diameter and with a partial reversal of the smoking-induced decrease in hyperemic brachial artery diameter (placebo/smoking session: from 4.68 +/- 0.13 mm to 4.53 +/- 0.15 mm, sildenafil/smoking session: from 4.72 +/- 0.12 mm to 4.64 +/- 0.13 mm, ie, improvement of 1.5%, P < .005). CONCLUSIONS: The present study shows, for the first time, that type 5 PDE inhibition with sildenafil abrogates the smoking induced acute decrease in FMD of the brachial artery. These findings may have clinical implications given the detrimental consequences of smoking and the strategic role of normal endothelial function. PMID- 15533732 TI - An association study in essential hypertension using functional polymorphisms in lymphotoxin-alpha gene. AB - BACKGROUND: Lymphotoxin-alpha (LTA), a member of the tumor necrosis factor family, is a cytokine produced by lymphocytes. The substance LTA mediates a wide variety of inflammatory, immunostimulatory, and antiviral responses. In 2002, LTA was identified as a major risk factor for myocardial infarction (MI) in Japanese individuals, in a large-scale case-control study using 92,788 gene-based single nucleotide polymorphism (SNP) markers in the whole human genome. Essential hypertension (EH) is thought to be a multifactorial disorder involved in endothelial dysfunction and atherosclerosis. Although hypertension is one of the greatest risk factors for MI, there have been no reports estimating the association between EH and LTA. The aim of the present study was to evaluate the association between EH and the LTA gene. METHODS: In the present study, we assessed the association between EH and SNP and haplotypes of the LTA gene in a case-control study of 202 EH patients and 217 age-matched normotensive control subjects. RESULTS: The overall distribution of genotypes for each SNP did not significantly differ between the two groups. Furthermore, the haplotype analysis revealed no association between the EH and normotensive groups. CONCLUSIONS: Polymorphisms of the LTA gene were not associated with EH. This finding suggests differences in genetic backgrounds between EH and MI. PMID- 15533733 TI - Effects of valsartan and nifedipine coat-core on systemic arterial stiffness in hypertensive patients. AB - BACKGROUND: The aim of this study was to compare the effects of long-term antihypertensive treatment on brachial-ankle pulse wave velocity (PWV), a marker of systemic arterial stiffness, between angiotensin II receptor blocker valsartan and long-acting calcium channel blocker nifedipine coat-core. METHODS: Forty-one patients (54 +/- 3 years of age, 20 men and 21 women) with essential hypertension (155 +/- 3/95 +/- 3 mm Hg) were randomly allocated to the treatment with valsartan (80 mg once daily) or nifedipine coat-core (20 mg once daily). Brachial ankle PWV and 24-h ambulatory blood pressures (BP) were measured before and 3 months after treatment. RESULTS: Baseline data did not differ between the valsartan and nifedipine groups. The PWV also was similar between the groups (1669 +/- 65 v 1622 +/- 64 cm/sec). Three months of treatment similarly reduced resting systolic and diastolic BP (nifedipine, -18.4 +/- 4.2/-11.9 +/- 2.7 mm Hg; valsartan, - 17.4 +/- 3.3/-9.8 +/- 2.1 mm Hg, all P < .001). The PWV was significantly reduced compared with baseline values in the valsartan group (-195 +/- 42 cm/sec, P < 0.001) but not in the nifedipine group (-69 +/- 40 cm/sec, NS). The 24-h mean heart rate increased in the nifedipine group but remained unchanged in the valsartan group, although BP were similarly lowered for 24 h. A tachycardic response was associated with an increase or lesser reduction in PWV in a group treated with nifedipine (r = 0.584, P < .01). CONCLUSIONS: These data suggest that long-term treatment with valsartan could reduce arterial stiffness better than nifedipine-coat core. The favorable vascular effect of valsartan was due in part to its nonhypotensive effect. The expected decrease in arterial stiffness may be offset by reflex sympathetic activation in some patients treated with nifedipine. PMID- 15533734 TI - A randomized, double-blind, placebo-controlled trial of casein protein hydrolysate (C12 peptide) in human essential hypertension. AB - BACKGROUND: Many patients seek complementary medicine treatments like neutraceuticals for common conditions such as hypertension. METHODS: We conducted a placebo-controlled prospective randomized crossover study in 10 hypertensive subjects to determine whether a single dose of a hydrolysate of bovine milk protein (designated C12 peptide; low and high dose), either alone or combined with alginic acid (low and high dose), reduced daytime blood pressure (BP), as determined by ambulatory BP monitoring. RESULTS: Within the five treatment regimens a significant reduction of 9.2 +/- 3.2 mm Hg in systolic BP at h 6 compared with h 2 occurred on the higher dose of alginic acid (1754 mg) combined with C12 (P = .02). The C12 peptide with the higher dose of alginic acid also showed a significant reduction of 6.0 +/- 2.0 mm Hg in diastolic BP at h 6 compared with h 2 (P = .015). CONCLUSIONS: These preliminary data are encouraging and deserve testing in a larger and longer treatment trial. PMID- 15533735 TI - Validation of the transfer function technique for generating central from peripheral upper limb pressure waveform. AB - Central aortic pressure waveforms can be calculated from the radial artery pressure waveform using a generalized transfer function to correct for pressure wave distortion in the upper limb. Although validated to standards conventionally applied, reservations are still expressed on use of this process, because of the relatively small number of patients from whom appropriate invasive data were obtained. The study described here supplemented such data with noninvasive data obtained from carotid and radial artery tonometry in 439 patients and normal subjects. The carotid-radial artery transfer function was similar to the aortic radial when allowance was made for wave travel from aorta to carotid artery. The carotid-radial transfer function was identical in male and female individuals, was similar at different arterial pressures and in mature adults. Differences are relatively small, are seen at frequencies where central pressure wave components are small and are similar to those seen with vasodilator agents in invasive studies. Findings provide further support for use of a generalized transfer function to calculate aortic from upper limb pressure and conform with previously established views on vascular impedance. PMID- 15533736 TI - Beyond hypertension toward guidelines for cardiovascular risk reduction. AB - BACKGROUND: Most current clinical guidelines focus primarily on the management of individual cardiovascular risk factors, such as high blood pressure (BP), hypercholesterolemia, or diabetes. A more appropriate clinical approach to reducing cardiovascular disease risk would be based on a comprehensive evaluation of risk profile, and accurate stratification of global (absolute) risk in individual patients. We propose that global risk should be used as the main determinant of whom to treat, how to treat, and how much to treat. METHODS: In this article we use a series of case studies to demonstrate the implications of replacing the traditional "single risk factor-based" approach to managing hypertension by one based on global risk assessment. In some situations patients with mildly elevated BP levels would not be recommended for antihypertensive drug treatment whereas others with lower BP would be treated, depending upon the entire risk profile. CONCLUSION: We propose to replace the single risk factor based approach with the assessment of global cardiovascular risk, both in the clinical management of individual patients and in guidelines. PMID- 15533737 TI - Blood pressure variability in hypertension: a possible cardiovascular risk factor. PMID- 15533738 TI - Plagiarism and scientific misconduct. PMID- 15533740 TI - "Reliability" of scientific reporting: a question of ethics. PMID- 15533742 TI - Strabismus following posterior segment surgery. AB - Persistent diplopia following posterior segment surgery is not uncommon. There are several causes of postoperative strabismus, some of which are preventable. Knowledge of the possible causes, proper evaluation, and appropriate treatment can eliminate or at least reduce the morbidity from this unwelcome postoperative surprise. PMID- 15533743 TI - Elevated intraocular pressure following vitreoretinal surgery. AB - Elevated intraocular pressure is a common occurrence following vitreoretinal surgery. The use of silicone oil or intravitreal gas in conjunction with pars plana vitrectomy increases the likelihood of postoperative intraocular pressure elevation. Fortunately, the pressure elevation is usually transient and managed medically. In certain instances, surgical intervention is warranted. Therapeutic intervention should be tailored to the individual patient based on the mechanism of pressure elevation. PMID- 15533744 TI - Corneal and conjunctival changes after posterior segment surgery. AB - Significant corneal compromise can occur subsequent to vitreoretinal surgery, especially in diabetic corneas. Associated factors include the operating lens systems used, irrigating solutions, preoperative lens and anterior capsule status, and the use of adjunctive agents such as intraocular gasses or silicone oil. Corneal, conjunctival, and ocular surface complications can also occur after scleral buckling procedures, often related to buckle extrusion or infection. PMID- 15533745 TI - Refractive changes after posterior segment surgery. AB - Retinal surgery can induce significant refractive errors. These errors include spherical changes caused by alterations in axial length after scleral buckle placement, astigmatic changes induced by a scleral buckle or pars plana vitrectomy, and focal alterations in corneal curvature that can significantly limit postoperative visual acuity. The adjunctive use of silicone oil can impose alterations directly, by the oil's interaction with the other refractive elements of the eye, and indirectly, through its effects on intraocular lens power calculations for subsequent cataract surgery. PMID- 15533746 TI - Anterior segment inflammation and hypotony after posterior segment surgery. AB - Recent advances in the instrumentation and surgical techniques for posterior segment surgery permit better anatomic and functional outcomes. Nevertheless, these procedures may be associated with complications involving the anterior segment, including postoperative inflammation and hypotony. One must differentiate infectious from noninfectious inflammation to initiate appropriate therapy promptly. Hypotony is a frustrating problem because it is difficult to reverse. The exclusion of eyes with other known causes of hypotony is important before establishing the putative mechanism of tractional ciliary body detachment caused by epiciliary proliferative tissue. PMID- 15533747 TI - Anterior ischemia after posterior segment surgery. AB - Anterior segment ischemia is a rare complication of posterior segment surgery with a broad spectrum of presentations. Most frequently, it follows a mild self limited course. Cases that are more prominent usually result from a co-incidence of precipitating factors. Recognition of precipitating factors and appropriate action can avoid anterior segment ischemia. Management includes nonspecific steps, such as topical medications (ie, corticosteroids and cycloplegics) and face-mask and eye-mask oxygen delivery, as well as specific steps related to the underlying cause (eg, removal of an encircling scleral buckle). PMID- 15533748 TI - Anterior segment complications associated with scleral buckling. AB - This article discusses the early and late postoperative complications of scleral buckling involving the anterior segment of the eye. Emphasis is placed on the incidence, pathogenesis, clinical presentation, and treatment of each complication. Preventive measures are also discussed. PMID- 15533749 TI - Cataracts associated with posterior segment surgery. AB - Although cataract is widely described among the complications of vitrectomy, the precise relationship between vitrectomy and the development of lens opacity is not completely understood. This article presents an extensive literature review and the authors' personal experience with this complication. PMID- 15533750 TI - Anterior segment complications related to vitreous substitutes. AB - Vitrectomy is the most common surgical technique performed in eyes with vitreoretinal disease. The development and widespread use of vitreous substitutes has revolutionized vitreoretinal surgery and improved anatomic and visual results. The three most common types of vitreous substitutes available in North America include silicone oil, intraocular gas, and perfluorocarbon liquid. Each of these agents has unique properties, allowing completely different roles of the agents in vitreoretinal surgery. The physical properties, clinical indications, and potential complications of these agents are described. PMID- 15533751 TI - Retinopathy of prematurity and anterior segment complications. AB - The management of retinopathy of prematurity (ROP) has changed over the last 20 years. Screening and treatment protocols have been established to provide timely therapy to infants at risk. Cryotherapy, laser photoablation, vitrectomy, and scleral buckling have proven effective but also have inherent risks of complications. The anterior segment complications related to ROP therapy are presented herein. PMID- 15533752 TI - Anterior segment complications following periocular and intraocular injections. AB - Significant anterior segment complications can occur following periocular or intraocular injections. These complications may be intraoperative or postoperative. They vary with different procedures and may be secondary to the surgical procedure or related to drug toxicity. Cataract and increased intraocular pressure are the most frequent complications. Other less commonly reported complications include hypotony, hyphema, corneal decompensation, strabismus, and cosmetic complications. PMID- 15533755 TI - Heparan sulfate proteoglycans and heparanase--partners in osteolytic tumor growth and metastasis. AB - This review summarizes a series of studies demonstrating that heparan sulfate proteoglycans act to promote the growth and metastasis of myeloma and breast tumors, two tumors that home to, and grow within, bone. Much of the growth promoting effect of proteoglycans in these tumors may reside in the shed form of syndecan-1 that acts to favorably condition the tumor microenvironment. Moreover, the interplay between heparan sulfate and the extracellular enzyme heparanase-1 also has important regulatory implications. Recent studies indicate that the activity of heparanase, which likely releases heparin sulfate-bound growth factors and generates highly active heparan sulfate fragments, also promotes growth and metastasis of myeloma and breast tumors. Understanding the role of heparan sulfate and heparanase in the regulation of tumor behavior may lead to new therapeutic approaches for treating cancer. PMID- 15533754 TI - The emergence of integrins: a personal and historical perspective. PMID- 15533756 TI - Lysophosphatidic acid inhibits TGF-beta-mediated stimulation of type I collagen mRNA stability via an ERK-dependent pathway in dermal fibroblasts. AB - Lysophosphatidic acid (LPA) is a serum-derived pleiotropic mediator with a potential role in wound repair. Since extracellular matrix (ECM) deposition is a critical part of wound healing, this study was designed to examine whether LPA is involved in ECM regulation. Using human dermal fibroblasts, we demonstrate that LPA counteracts transforming growth factor-beta (TGF-beta) stimulation of type I collagen mRNA and protein. This factor elicits its inhibitory effects at the posttranscriptional level via destabilization of type I collagen mRNA. Furthermore, using the mitogen-activated protein kinase kinase (MEK) inhibitor PD98059, we show that the extracellular signal-regulated kinase (ERK) pathway is a negative regulator of the TGF-beta-induced stabilization of type I collagen mRNA, and that the activation of the ERK pathway by LPA mediates their inhibitory effects on collagen production. In conclusion, this study describes a novel function for LPA as an antagonist of TGF-beta induced ECM deposition. These findings may be relevant to physiologic wound repair and may be useful in designing therapeutic agents to prevent excessive scarring. PMID- 15533757 TI - Osmotic stress regulates the anticoagulant efficiency of dermatan sulfate. AB - Glycosaminoglycans (GAGs) in pericellular and interstitial spaces help to maintain local water homeostasis and blood coagulation balance. This study explored whether dehydrating microenvironment conditions influence dermatan sulfate's (DS) anticoagulant activity. Water transfer during antithrombin activation by dermatan sulfate was measured using osmotic stress techniques. Anticoagulant activity was determined from the change in the rate of coagulation factor Xa (fXa) inhibition. Osmotic stress accelerated reaction rates, indicating water transfer from reactants to bulk. The net volume transferred, measured using osmotic probes similar in size to the reacting proteins, was approximately 2500 mol of water per mole of fXa inhibited. The reaction efficiency, V(sat)/K 1/2 (rate at saturation/concentration resulting in half-maximal rates), determined in titrations with monosulfated dermatan sulfate and disulfated dermatan sulfate (DDS), were 4x10(4) and 2x10(5) M-1 s-1 under osmotic stress and in the presence of calcium, corresponding to 34- and 81-fold increases over efficiency measured under standard conditions. These results indicate that dermatan sulfate can contribute significantly to antithrombin activation, and that in dehydrating environments and depending of ionic conditions, its anticoagulant efficiency can exceed that of heparan sulfate (HS). PMID- 15533758 TI - Colocalization of dentin matrix protein 1 and dentin sialoprotein at late stages of rat molar development. AB - Dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) are acidic proteins found in the extracellular matrices of bones and teeth. Recent data from gene knockouts, along with those of gene mutations, indicate that these two phosphoproteins are critical for bone and tooth development and/or maintenance. However, the precise functions of the two proteins have not been elucidated. In order to gain insights into their functions in tooth formation, we performed systematic, comparative investigations on the immunolocalization of DMP1 and dentin sialoprotein (DSP, a cleaved fragment of DSPP), using the rat first molar at different developmental stages as a model. Immunohistochemistry (IHC) was performed with specific, monoclonal antibodies against the COOH-terminal fragments of DMP1 and against DSP. In 1-day- and 1-week-old rats, weak immunoreactions for DMP1 were observed in dentinal tubules while stronger reactions for DSP were seen in the tubules and predentin. In rats older than 2 weeks, immunoreactions for DMP1 were found in dentinal tubules, predentin and odontoblasts. In 5-week- and 8-week-old rats, strong immunoreactions for DMP1 were widely distributed in odontoblasts and predentin. The distribution pattern of DSP was strikingly similar to that of DMP1 after 2 weeks and the localization of each was distinctly different from that of bone sialoprotein (BSP). The unique colocalization of DMP1 and DSPP in tooth development suggests that the two proteins play complementary and/or synergistic roles in formation and maintenance of healthy teeth. PMID- 15533759 TI - Altered patterns and synthesis of extracellular matrix macromolecules in early osteoarthritis. AB - The synthesis and contents of extracellular non-collagenous matrix macromolecules was studied in early and late human osteoarthritic (OA) cartilage obtained at surgery for sarcomas in the lower extremities (normal and early OA) or for total knee replacement (late stage OA). The early OA samples were those that had some fibrillation in the joint by visual examination. One group had fibrillation in the area sampled and the other group had no fibrillation. Cartilage was taken from the same topographical area on the medial femoral condyle in all the samples, labeled with [3H]leucine and [35S]sulfate for 4 h at 37 degrees C and extracted with 4 M guanidine-HCl. Analysis of the extracts showed that the total amount of proteoglycans relative to hydroxyproline content was higher in the early and late OA than in the normal cartilage. These proteoglycans showed a relatively lower [35S]sulfate incorporation into GAG chains and a higher [3H]leucine incorporation. The pattern of newly synthesized proteins was altered similarly in early and late OA. Notably, synthesis of cartilage oligomeric matrix protein (COMP), fibronectin, and cartilage intermediate layer protein (CILP) was increased, also reflected in their abundance as determined by enzyme-linked immunosorbent assay (ELISA). Collagen synthesis appeared significantly increased only in the late stage OA. The observed altered composition and pattern of biosynthesis indicate that the joint undergoes metabolic alterations early in the disease process, even before there is overt fibrillation of the tissue. The early OA samples studied appear to represent two distinct groups of early lesions in different stages of the process of cartilage deterioration as shown by their differences in relative rates of synthesis and abundance of proteins. PMID- 15533760 TI - Molecular mechanisms of platinum resistance: still searching for the Achilles' heel. AB - The platinum compounds cisplatin and carboplatin are commonly used in cancer chemotherapy. However, tumors frequently develop resistance to these compounds, significantly decreasing their usefulness in the clinic. In the past few years, basic research has unraveled novel and unexpected mechanisms for the development of platinum resistance. For example, it has been reported that MUC1 expression and particularly the localization of its C-terminal subunit to the mitochondria may affect cisplatin resistance. Another recent finding suggests that cisplatin damage may activate DNA-dependent protein kinase (DNA-PK) to initiate a death signal that can be transmitted to neighboring cells through gap junctions, adding to a growing belief that the interactions of cancer cells with their surroundings may be important to the outcome of chemotherapy. While most clinical efforts have focused on identifying alternative regimens for drug-resistant cancer, it might be possible to exploit our knowledge of the mechanism of platinum resistance to specifically reverse resistance and increase platinum efficacy. The strategy of drug resistance reversal therapy (DRRT) may have significant impact on our approaches to the treatment and management of drug-resistant tumors. PMID- 15533761 TI - Artemisinins: mechanisms of action and potential for resistance. AB - Artemisinins form the most important class of antimalarial currently available, particularly because they are effective against parasites resistant to almost all the other classes. Their mechanism of action is controversial. Some aspects of this controversy are reviewed here. Whilst there is no clinical resistance yet identified to artemisinins, the potential to examine the relationship between polymorphisms in PfATP6 (a target of artemisinins) in multidrug resistant isolates of Plasmodium falciparum, is also discussed. PMID- 15533762 TI - Prediction of drug sensitivity and drug resistance in cancer by transcriptional and proteomic profiling. AB - The oncologist's challenges, particularly with advanced cancers, are (a) how to predict tumor response to a given drug or regimen; (b) how to predict which tumors of identical histology will remain indolent and which will be likely to progress; and (c) how to determine the appropriate timing of the emergence of drug-resistant cancer cells and hence switch to appropriate therapy. These issues are still unresolved; current clinical practice is hampered by the complexity and heterogeneity of anti-tumor drug resistance where multiple cellular, tumor microenvironment and host factors operate simultaneously. The rapid accumulation of genomic and proteomic databases for complex biological systems, such as cancer, together with advances in technology platforms, have paved the way to an increased molecular understanding and prediction of antitumor drug response. The complex phenotype of drug resistance can now be dissected and specific, clinically relevant markers pinpointed. Several microarray studies of genetic patterns from untreated and pre-treated cancers have provided "fingerprints" that can predict response to therapeutics. Nevertheless, such approaches require further validation in experimental models and in large clinical trials before their routine clinical use. Moreover, comparative transcriptional profiling alone is unlikely to predict drug sensitivity/resistance, a dynamic process where protein phosphorylation, protein trafficking, and protein-protein interactions with secondary effectors play key roles in the fate of cancer cells following therapeutic stress. Functional proteomics is potentially more predictive, but still faces technical challenges with regards to sampling, tumor heterogeneity, and lack of standardized methodologies. These obstacles are surmountable with current concerted research efforts and availability of powerful high-throughput genomic and proteomic instrumentations, and thus approaches to predict and overcome drug resistance could be rationalized. PMID- 15533763 TI - Leishmaniasis: drugs in the clinic, resistance and new developments. AB - The control of Leishmania infections relies primarily on chemotherapy. The arsenal of drugs available for treating Leishmania infections is limited and includes pentavalent antimonials, pentamidine, amphotericin B, miltefosine, fluconazole and few other drugs at various stages of their development process. In this review, we will discuss the latest results regarding resistance mechanisms to drugs used in the clinic against Leishmania infections. PMID- 15533764 TI - Epigenetic silencing mediated by CpG island methylation: potential as a therapeutic target and as a biomarker. AB - Many genes become transcriptionally silenced during the development of cancer. As well as affecting disease progression, gene silencing has the potential to influence drug resistance and clinical outcome following therapy. In addition to silencing due to gene mutations, covalent epigenetic modifications such as DNA hypermethylation and histone post-translational modifications are associated with transcriptional inactivation of many genes and are an important early event during carcinogenesis and tumour development. Aberrant methylation of CpG islands in promoters is associated with transcriptional inactivation of genes involved in all aspects of tumour development. Genes involved in key DNA damage response pathways, such as cell cycle control, apoptosis signalling and DNA repair, can frequently become methylated and epigenetically silenced in tumours. This may lead to differences in intrinsic sensitivity of tumours to chemotherapy, depending on the specific function of the gene inactivated. Furthermore, it is proposed that chemotherapy itself can exert a selective pressure on epigenetically silenced drug sensitivity genes present in subpopulations of cells, leading to acquired chemoresistance. Since the DNA sequence of epigenetically inactivated genes are not mutated but rather subject to reversible modifications via DNA methyltransferases (DNMTs) or histone modification, it is possible to reverse silencing using small molecule inhibitors. Such compounds show anti-tumour activity and can increase the sensitivity of drug resistant preclinical tumour models. Clinical trials of epigenetic therapies are now underway. Epigenetic profiling, using DNA methylation and histone analysis, will provide guidance on optimisation of these therapies with conventional chemotherapy and will help identify patient populations who may particularly benefit from such approaches. PMID- 15533765 TI - Withdrawing antimalarial drugs: impact on parasite resistance and implications for malaria treatment policies. AB - Malaria continues to be a leading cause of death in the tropics, taking the heaviest toll on children in Africa, where drug resistant Plasmodium falciparum has led to rising malaria mortality. High rates of chloroquine resistance prompted many countries in Africa to switch to alternative therapies to treat malaria. Parasites carrying mutations that render them chloroquine resistant may lose their survival advantage with the removal of chloroquine drug pressure. Alternatively, organisms may have undergone compensatory mutation that provides a survival advantage even in the absence of drug pressure. Decreasing drug resistant malaria has been reported following discontinuation of antimalarial drugs. However, most such reports are limited by the incomplete removal of chloroquine drug pressure, unreliable in vitro susceptibility assays and/or small, poorly described study populations. In Africa, Malawi was the first country to switch from chloroquine to sulfadoxine-pyrimethamine for the first line treatment of malaria. An effective campaign to end chloroquine use provided an excellent opportunity to study the natural history of drug resistance following the reduction of drug pressure. The finding that drug resistance decreases with the removal of drug pressure could provide a new paradigm for malaria treatment policies in Africa. PMID- 15533766 TI - Vascular remodeling and clinical resistance to antiangiogenic cancer therapy. AB - When first conceived, antiangiogenic therapy for cancer offered the possibility of universal efficacy, low toxicity, and little possibility of resistance. Blockade of the vascular endothelial growth factor (VEGF) pathway has yielded the most promising results both in animal models and in patients. However, resistance to VEGF blockade has been found even when given in combination with chemotherapy or other antiangiogenic agents. This resistance is associated with remodeled vasculature and with increased expression of angiogenic factors, such as PDGF-B and angiopoietin-1, which may contribute to vessel stabilization. Future efforts must be directed towards the identification of factors associated with vascular remodeling in order to improve the efficacy of antiangiogenic therapy. PMID- 15533767 TI - Candida biofilm resistance. AB - Device-related infections in most nosocomial diseases can be traced to the formation of biofilms (microbial communities encased within polysaccharide-rich extracellular matrix) by pathogens on surfaces of these devices. Candida species are the most common fungi isolated from these infections, and biofilms formed by these fungal organisms are associated with drastically enhanced resistance against most antimicrobial agents. This enhanced resistance contributes to the persistence of this fungus despite antifungal therapy. Candida biofilms exhibit enhanced resistance against most antifungal agents, except echinocandins and lipid formulations of AmB. The expression of drug efflux pumps during the early phase of biofilm formation and alterations in membrane sterol composition contribute to resistance of these biofilms against azoles. Metabolic dormancy and ECM do not appear to contribute to resistance, although in a mixed-species biofilm, ECM does retard the diffusion of drugs across biofilm. These multifactorial mechanisms of resistance in fungal biofilms constitute a broad spectrum defense that is effective against many types of antifungal agents, and represent a common theme present across microbial biofilms. PMID- 15533769 TI - Clinical endocrinology and metabolism. Development of gut endocrine cells. AB - During development, the definitive endoderm differentiates into several gastrointestinal epithelial lineages, including enteroendocrine cells. The enteroendocrine lineage consists of at least 15 different cell types that are categorized based on their morphology, location and peptide hormone expression. The mechanisms regulating enteroendocrine cell differentiation are likely to be critical not only in embryonic development, but also during the constant renewal of gut epithelia in the adult. The identification of transcription factors and regulatory DNA elements required for cell type-specific gene expression in various endocrine cell types has broadened our understanding of the regulatory networks controlling the spatial and temporal activation of enteroendocrine differentiation programs. This chapter will review recent studies of transcription factors during enteroendocrine cell differentiation, with a focus on the central role for the Notch signaling pathway in enteroendocrine cell fate decisions. PMID- 15533770 TI - Clinical endocrinology and metabolism. Receptors for gut peptides. AB - Most gut peptides exert their effects through G protein-coupled receptors, a family of about 700 membrane proteins, 87 of which are presently known to have peptide ligands. Three additional gut peptide receptors are not G protein-coupled receptors but regulate intracellular cyclic GMP accumulation. The aim of this review is to illustrate how the sequencing of the human genome and other recent advances in genomics has contributed to our understanding of the role of peptides and their receptors in gastrointestinal function. Recent discoveries include the identification of receptors for the peptides motilin and neuromedin U, and new physiological ligands for the PTH2 receptor, the CRF(2) receptor and the growth hormone secretagogue receptor. Knockout mice lacking specific peptide receptors or their ligands provide informative animal models in which to determine the functions of the numerous peptide-receptor systems in the gut and to predict which of them may be the most fruitful for drug development. Some peptide receptor signalling systems may be more important in disease states than they are in normal physiology. For example, substance P, galanin, bradykinin and opioids play important roles in visceral pain and inflammation. Other peptides may have developmental roles: for example, disruption of endothelin-3 signalling prevents the normal development of the enteric nervous system and contributes to the pathogenesis of Hirschsprung disease. PMID- 15533771 TI - Clinical endocrinology and metabolism. Gut endocrine tumours. AB - Endocrine tumours of the gastrointestinal tract and pancreas may present at different disease stages with either hormonal or hormone-related symptoms/syndromes, or without hormonal symptoms. They may occur either sporadically or as part of hereditary syndromes. In the therapeutic approach to a patient with these tumours, excessive hormonal secretion and/or its effects should always be controlled first. A team approach is needed to achieve a balanced opinion on the use of the different therapeutic options in patients with these tumours. PMID- 15533772 TI - Clinical endocrinology and metabolism. Regulation of energy homeostasis by peripheral signals. AB - The increased incidence of obesity makes it imperative to understand the regulation of food intake and body weight. We review the signals that interact with the brain to control energy homeostasis, i.e. energy intake and expenditure. Three broad categories can be distinguished. Signals generated in the gastrointestinal tract during meals ('satiety' signals, e.g. cholecystokinin) elicit satiation and contribute to stopping the meal. The potency of these acutely acting signals must be increased if they are to be used therapeutically. Hormonal signals whose secretion is proportional to body fat (adiposity signals, leptin and insulin) robustly reduce food intake and body weight by directly stimulating receptors locally in the brain. Therapeutic applications will have to find ways to circumvent the systemic actions of these hormones, targeting only the brain. Satiety and adiposity signals interact with neuronal circuits in the brain that utilize myriad neurotransmitters to cause net catabolic or anabolic responses. Considerable effort is being directed towards finding ways to intervene in specific circuits to help accomplish weight loss. PMID- 15533773 TI - Clinical endocrinology and metabolism. Ghrelin, a novel growth-hormone-releasing and appetite-stimulating peptide from stomach. AB - Recent identification of novel appetite-regulating hormones has revealed the complex interactions of these humoral factors in the regulation of feeding behavior in mammals. One of these hormones, ghrelin, a natural ligand of the orphan receptor GHS-R, purified from stomach, is able to stimulate growth hormone release from pituitary cells. Ghrelin is a 28 amino acid peptide containing an n octanoylated serine 3 residue that is essential for its activity. Ghrelin stimulates appetite by acting on the hypothalamic arcuate nucleus, the region known to control food intake. As an orexigenic peptide, ghrelin is therefore an endogenous regulator of feeding behavior from the peripheral tissues to the central nervous system. PMID- 15533774 TI - Clinical endocrinology and metabolism. Glucagon-like peptide-1 and glucagon-like peptide-2. AB - The glucagon-like peptides (glucagon-like peptide-1 (GLP-1) and glucagon-like peptide-2 (GLP-2)) are released from enteroendocrine cells in response to nutrient ingestion. GLP-1 enhances glucose-stimulated insulin secretion and inhibits glucagon secretion, gastric emptying and feeding. GLP-1 also has proliferative, neogenic and antiapoptotic effects on pancreatic beta-cells. More recent studies illustrate a potential protective role for GLP-1 in the cardiovascular and central nervous systems. GLP-2 is an intestinal trophic peptide that stimulates cell proliferation and inhibits apoptosis in the intestinal crypt compartment. GLP-2 also regulates intestinal glucose transport, food intake and gastric acid secretion and emptying, and improves intestinal barrier function. Thus, GLP-1 and GLP-2 exhibit a diverse array of metabolic, proliferative and cytoprotective actions with important clinical implications for the treatment of diabetes and gastrointestinal disease, respectively. This review will highlight our current understanding of the biology of GLP-1 and GLP-2, with an emphasis on both well-characterized and more novel therapeutic applications of these peptides. PMID- 15533775 TI - Clinical endocrinology and metabolism. Gastrin. AB - The gastric hormone gastrin stimulates gastric acid secretion and epithelial cell proliferation. Multiple active products are generated from the precursor, preprogastrin, including the well-characterized amidated gastrins acting at the cholecystokinin-2 (CCK-2, or gastrin-CCK(B)) receptor, and others that may be growth factors in a range of cancers. Plasma concentrations of the amidated gastrins are elevated as a consequence of gastrin-secreting tumours (gastrinomas) and in conditions in which the normal inhibition of the antral G-cell by acid is depressed, for example chronic atrophic gastritis and prolonged treatment with proton pump inhibitors. There may also be increased gastrin release in Helicobacter pylori infection. Provocative tests for the diagnosis of gastrinoma include the secretin and calcium infusion tests. Hypergastrinaemia is associated with enterochromaffin-like (ECL) cell proliferation; the factors that determine progression to ECL cell dysplasia and gastric ECL cell carcinoid tumours are discussed. Several strategies for inhibiting the effects of gastrin are under evaluation, and their potential application is discussed. PMID- 15533776 TI - Clinical endocrinology and metabolism. Cholecystokinin. AB - Cholecystokinin (CCK) is a peptide hormone discovered in the small intestine. Together with secretin and gastrin, CCK constitutes the classical gut hormone triad. In addition to gallbladder contraction, CCK also regulates pancreatic enzyme secretion and growth, intestinal motility, satiety signalling and the inhibition of gastric acid secretion. CCK is, however, also a transmitter in central and intestinal neurons. Notably, CCK is the most abundant neuropeptide in the human brain. Owing to difficulties in developing accurate assays, knowledge about CCK secretion in disease is limited. Available data indicate, however, that proCCK is expressed in certain neuroendocrine tumours and sarcomas, whereas the secretion of CCK is impaired in celiac disease and bulimia nervosa. Stimulation with exogenous CCK has proved useful in diagnostic tests of gallbladder and pancreatic diseases, as well as medullary thyroid carcinomas. PMID- 15533777 TI - Clinical endocrinology and metabolism. Glucose-dependent insulinotropic polypeptide/gastric inhibitory polypeptide. AB - The 42 amino acid polypeptide glucose-dependent insulinotropic polypeptide/gastric inhibitory polypeptide (GIP) is released from intestinal K cells in response to nutrient ingestion. Based on animal studies, the peptide was initially assumed to act as an endogenous inhibitor of gastric acid secretion. Later it was found that GIP is capable of augmenting glucose-stimulated insulin secretion, and subsequent studies provided evidence that, in humans, the peptide predominantly acts as an incretin hormone. A role for GIP in the regulation of lipid homeostasis and in the development of obesity has been inferred from different animal studies. While GIP strongly stimulates insulin release in healthy humans, the peptide has almost completely lost its insulinotropic effect in patients with type 2 diabetes. This is different from the actions of glucagon like peptide 1, which stimulates insulin secretion even in the later stages of type 2 diabetes. This suggests that a diminished insulinotropic effect of GIP may contribute to the pathogenesis of type 2 diabetes. This review will summarize the actions of GIP in human physiology and discuss its role in the pathogenesis of type 2 diabetes, as well as the therapeutic options derived from these findings. PMID- 15533778 TI - Clinical endocrinology and metabolism. The somatostatin neuroendocrine system: physiology and clinical relevance in gastrointestinal and pancreatic disorders. AB - Somatostatin is produced in enteroendocrine D cells and intrinsic neurons of the stomach, intestines and pancreas. Its physiologic actions are mediated primarily by somatostatin receptors type 2 and 5, and include the inhibition of secretion of most endocrine and exocrine factors. Diseases directly attributable to somatostatin excess or deficiency are rare, although there is a complex pathogenic relationship between persistent Helicobacter pylori infection and reduced somatostatin in chronic gastritis. Abundant somatostatin receptors on many neoplastic and inflammatory cells are the basis for sensitive in vivo imaging with radiolabeled somatostatin analogs and provide a therapeutic target. Current indications for somatostatin therapy include hormone-expressing neuroendocrine tumors, intractable diarrhea and variceal bleeding secondary to portal hypertension. Exciting advances are being made in the development of high affinity nonpeptide analogs with receptor-subtype selectivity and increased bioavailability. Somatostatin analogs coupled to high-energy radionuclides show promise as novel cytotoxic agents for certain metastatic tumors. PMID- 15533779 TI - Clinical endocrinology and metabolism. Potential clinical applications of vasoactive intestinal peptide: a selected update. AB - Neuropeptides are expressed in neurons innervating endocrine cells or in endocrine cells and cancer cells, and are released on site to act as hormones and growth factors. Vasoactive intestinal peptide (VIP) was first discovered in the early 1970s and has since become the area of research for many laboratories. VIP has a neuroendocrine role as it is intimately involved with the synthesis, secretion and action of other neuroendocrine hormones as well as cytokines and chemokines. Major outcomes of VIP downregulation encompass developmental and behavioral dysfunctions, including impaired diurnal rhythms. Overexpression of VIP has been associated with diarrhea and cancer, and overexpression of VIP receptors is associated with cancerous growth. This short review outlines some of the recent progress made in VIP research. PMID- 15533781 TI - BAC libraries and comparative genomics of aquatic chordate species. AB - The bacterial artificial chromosome (BAC) system is useful for creating a representation of the genomes of target species. The system is advantageous in that it can accommodate exogenous inserts that are very large (>100 kilobases, kb), thereby allowing entire eukaryotic genes (including flanking regulatory regions) to be encompassed in a single clone. The interest in BACs has recently been spawned by vast improvements in high throughput genomic sequencing such that comparisons of orthologous regions from different genomes (comparative genomics) are being routinely investigated, and comprise a significant component, of all major sequencing centers. In this review, we discuss the general principles of BAC cloning, the resources that are currently available, and some of the applications of the technology. It is not intended to be an exhaustive treatise; rather our goal is to provide a primer of the BAC technology in order to make readers aware of these resources and how they may utilize them in their own research programs. PMID- 15533782 TI - Induction of reversible hemolytic anemia in living zebrafish using a novel small molecule. AB - We used zebrafish to screen and identify small molecules that affect the process of vertebrate hematopoietic development. Zebrafish embryos were exposed to a library of 5000 synthetic compounds and screened for defects in primitive erythropoiesis. Here, we present the characterization of hemolytic anemia induced in zebrafish by the small molecule 5115318 (3-[5-methyl-furan 2-yl]-propionic acid N'-phenyl-hydrazide). This compound is capable of generating hemoglobin aggregates and Heinz bodies in red cells in vivo only. The induced anemia is reversible and treated fish recover in about 4 days. This study shows the feasibility of using zebrafish to screen for small molecules that can modulate the specific process of erythropoiesis. PMID- 15533783 TI - Developmental expression of aquaporin-3 in zebrafish embryos (Danio rerio). AB - Fish embryos have never been successfully cryopreserved because of the low permeability of cryoprotectants into the yolk. Recently, we used aquaporin-3 fused with a green fluorescent protein (AQP3GFP) to modify the zebrafish embryo, and demonstrated that the pores functioned physiologically. This increased the water and cryoprotectant permeability of the membranes. We have continued our work on AQP3-modified embryos and here we report their developmental expression of AQP3, the success of various culture media on their survival and development, and their reproductive success. The AQP3GFP expression begins within 30 m after the mRNA AQP3GFP injection into the yolk of the 1- to 4-cell embryo. This expression is distributed in the membranes throughout the blastoderm and the yolk syncytial layer within 24 h. It diminishes after 96 h. We found no difference in the survival or normal development of embryos from AQP3GFP or wild-type adults. Additionally, zebrafish embryos did not require special culture medium to survive after AQP3GFP modification. In fact, they survived best in embryo medium (ca. 40 mOsm). Embryos reared entirely in embryo medium had a higher percent survival and a higher percent normal development than those exposed to a high osmolality sucrose culture medium (ca. 330 mOsm). The mechanism whereby these embryos can maintain their internal osmolality in a hypoosmotic solution with water channels in their membranes is unknown. PMID- 15533785 TI - Investigating the morphology, function and genetics of cytotoxic cells in bony fish. AB - Bony fish (teleosts) possess multiple cytotoxic cell lineages that recognize and destroy virally infected and transformed cells. In general, these lineages parallel their functional equivalents in mammals and include neutrophilic granulocytes, macrophages, cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. These four cell types have been morphologically identified in multiple fish species but only limited information is available about their function. In contrast, much work has gone into examining the function of a fifth cytotoxic cell lineage, termed nonspecific cytotoxic cells (NCC), that has been referred to as the bony fish equivalent of NK cells. However, evidence suggesting that NCC do not represent the NK lineage has come through the development of multiple cytotoxic catfish cell lines that are morphologically and functionally similar to human NK cells and are distinct from NCC. In addition to characterizing cytotoxic cells from fish, recent work has identified the novel immune-type receptors (NITR) and cichlid killer leukocyte receptors (cKLR) that are structurally related to mammalian NK receptors and likely play a role in cytotoxic function in fish. This review summarizes the morphological and functional evidence for cytotoxic cells within bony fish and discusses future directions for examining cytotoxicity through genomics and transgenics. PMID- 15533784 TI - Cloning of anthozoan fluorescent protein genes. AB - Many cnidarians display vivid fluorescence under proper lighting conditions. In general, these colors are due to the presence of fluorescent proteins similar to the green fluorescent protein (GFP) originally isolated from the hydrozoan medusa Aequorea victoria (Cnidaria: Hydrozoa). To optimize the search for new fluorescent proteins (FPs), a technique was developed that allows for the rapid cloning and screening of FP genes without the need for a prior knowledge of gene sequence. Using this method, four new FP genes were cloned, a green from Montastraea cavernosa (Anthozoa: Scleractinia: Faviidae), a cyan from Pocillopora damicornis (Anthozoa: Scleractinia: Pocilloporidae), a cyan from Discosoma striata (Anthozoa: Corallimorpharia), and a red from a second Discosoma species. Two additional green FPs were cloned, one from M. cavernosa and one from its congener Montastraea faveolata, from purified cDNA using PCR primers designed for the first M. cavernosa green FP. Each FP has recognizable amino acid sequence motifs that place them conclusively in the GFP protein family. Mutation of these products using a low-stringency PCR protocol followed by screening of large numbers of bacterial colonies allowed rapid creation of mutants with a variety of characteristics, including changes in color, maturation time, and brightness. An enhanced version of the new red FP, DspR1+, matures faster at 30 degrees C than the commercially available DsRed but matures slower than DsRed at 37 degrees C. One of the M. cavernosa green FPs, McaG2, is highly resistant to photobleaching and has a fluorescence quantum yield approximately twice that of EGFP-1. PMID- 15533786 TI - Genetic, biochemical and evolutionary facets of Xmrk-induced melanoma formation in the fish Xiphophorus. AB - Certain interspecific hybrids of the fish Xiphophorus spontaneously develop melanoma induced by the derepression of the Xmrk oncogene. Xmrk is a recent duplicate of an orthologue of the mammalian epidermal growth factor receptor gene Egfr. In addition to a specific overexpression in melanoma, amino-acid substitutions in the extracellular domain leading to ligand-independent dimerisation and constitutive autophosphorylation are responsible for the tumorigenic potential of Xmrk. The Xmrk receptor induces several signal transduction pathways mediating cell proliferation and resistance to apoptosis and initiating dedifferentiation. Moreover, Xmrk upregulates the expression of the secreted protein osteopontin, inducing an autocrine loop possibly allowing invasion and survival in the dermis as a first step in malignancy. Hence, Xmrk is able to induce pathways essential for a transformed phenotype. Some of these events are equivalent to those found downstream of the mammalian Egfr, but others have clearly evolved differently or are specific for pigment cells. Xmrk is potentially hazardous, nonessential and located in a very unstable genomic region. Nevertheless, Xmrk has been maintained under purifying selection in divergent Xiphophorus species. Hence, Xmrk has probably a beneficial function under certain conditions. The analysis of this function is a major challenge for future research in the Xiphophorus model. PMID- 15533787 TI - Structural organization, mapping, characterization and evolutionary relationships of CDKN2 gene family members in Xiphophorus fishes. AB - Xiphophorus fishes and their hybrids are used as models for the study of melanoma and other diseases. The cyclin-dependent kinase inhibitor gene family in humans is comprised of four members, including CDKN2A (P16), and dysregulation of this gene is implicated in numerous neoplasms including melanomas. We have investigated the status of the gene family in the southern platyfish X. maculatus. Xiphophorus harbors at least two such loci, which we now term CDKN2A/B and CDKN2D. Both loci map to Xiphophorus linkage group 5, a genomic area that has long been known to harbor the DIFF tumor suppressor locus. Within this report, we report on the complete cloning, genomic exon/intron boundary delineation, linkage mapping and expressional characteristics of Xiphophorus CDKN2D. We also compare and contrast this expression to that of the previously isolated CDKN2AB locus in normal and neoplastic tissues derived from non-hybrid and hybrid fishes. The hypothetical evolutionary relationships of gene family members and their involvement in melanoma is evaluated. In comparison to CDKN2A/B, the RNA expression of Xiphophorus CDKN2D differs in normal tissues and is not associated with melanotic/pathologic tissues, confirming functional divergence between obvious homologues. PMID- 15533788 TI - DNA repair in hybrid fish of the genus Xiphophorus. AB - The genus Xiphophorus is an important vertebrate model for investigating the etiology and genetics of both spontaneous and induced cancers. Xiphophorus are comprised of 23 species most of which can be crossed to produce fertile interspecies hybrid progeny. The Xiphophorus gene map is well developed and allows genetic associations to be studied among cohorts of progeny derived from backcrossing interspecies hybrid animals to one of the parental strains. In interspecies cross-progeny from select Xiphophorus backcrosses, ionizing radiation, ultraviolet light (UVB), and exposure to methylnitrosourea (MNU) have all been shown to induce tumors. Induced tumor types represented in various models include melanoma, fibrosarcoma, schwannoma, retinoblastoma, etc. The well established backcross hybrid genetics make Xiphophorus fish an excellent system to study the contribution of DNA repair capability to induced tumorigenesis. DNA repair pathways represent multigenic traits that must be tightly regulated to insure genome fidelity. Herein we review initial DNA repair studies that assess repair capacities among different Xiphophorus species and interspecies hybrids. Assessment of both base excision repair (BER) and nucleotide excision repair (NER) have yielded consistent results indicating reduced DNA repair function in hybrid fish tissues. These data provide molecular support for potential reduced fitness in hybrid fish under conditions of environmental stress and may present a plausible explanation for absence of interspecies hybridization in sympatric environments. In addition, they support the role of direct DNA damage and its repair in the initiation of tumors in Xiphophorus hybrids. PMID- 15533789 TI - Gene structure, purification and characterization of DNA polymerase beta from Xiphophorus maculatus. AB - Cloning of the Xiphophorus maculatus Polbeta gene and overexpression of the recombinant Polbeta protein has been performed. The organization of the XiphPolbeta introns and exons, including intron-exon boundaries, have been assigned and were found to be similar to that for human Polbeta with identical exon sizes except for exon XII coding for an additional two amino acid residues in Xiphophorus. The cDNA sequence encoding the 337-amino acid X. maculatus DNA polymerase beta (Polbeta) protein was subcloned into the Escherichia coli expression plasmid pET. Induction of transformed E. coli cells resulted in the high-level expression of soluble recombinant Polbeta, which catalyzed DNA synthesis on template-primer substrates. The steady-state Michaelis constants (Km) and catalytic efficiencies (kcat/Km) of the recombinant XiphPolbeta for nucleotide insertion opposite single-nucleotide gap DNA substrates were measured and compared with previously published values for recombinant human Polbeta. Steady-state in vitro Km and kcat/Km values for correct nucleotide insertion by XiphPolbeta and human Polbeta were similar, although the recombinant Xiphophorus protein exhibited 2.5-7-fold higher catalytic efficiencies for dGTP and dCTP insertion versus human Polbeta. In contrast, the recombinant XiphPolbeta displayed significantly lower fidelities than human Polbeta for dNTP insertion opposite a single-nucleotide gap at 37 degrees C. PMID- 15533790 TI - DNA polymerase beta mRNA and protein expression in Xiphophorus fish. AB - Herein we report Xiphophorus DNA polymerase beta (XiphPolbeta) mRNA and protein expression levels in brain, liver, gill, and testes tissues from Xiphophorus maculatus, Xiphophorus helleri, and Xiphophorus couchianus parental line fish and two different tumor-bearing Xiphophorus interspecies hybrids. Polymerase beta protein levels in the Xiphophorus tissues were measured by Western blot, and mRNA was measured with a quantitative real time RT-PCR method which employed cRNA construction to produce accurate calibration curves. We found significant differences in both mRNA and protein levels between the tumor-bearing hybrid animals and the three parental species. However, there were no significant differences in either mRNA levels or protein expression observed between the parental species. Thus, interspecies hybridization results in dysregulation of Polbeta expression and this may manifest a modulation in DNA repair capability and susceptibility to latent tumorigenesis. PMID- 15533791 TI - The use of mature zebrafish (Danio rerio) as a model for human aging and disease. AB - Zebrafish (Danio rerio) have been extensively utilized for understanding mechanisms of development. These studies have led to a wealth of resources including genetic tools, informational databases, and husbandry methods. In spite of all these resources, zebrafish have been underutilized for exploring pathophysiology of disease and the aging process. Zebrafish offer several advantages over mammalian models for these studies, including the ability to perform saturation mutagenesis and the capability to contain thousands of animals in a small space. In this review, we will discuss the use of mature zebrafish as an animal model and provide specific examples to support this novel use of zebrafish. Examples include demonstrating that clinical pathology can be performed in mature zebrafish and that age-associated changes in heat shock response can be observed in zebrafish. These highlights demonstrate the utility of zebrafish as a model for disease and aging. PMID- 15533792 TI - Using zebrafish to study the complex genetics of glaucoma. AB - The overall goal of this review is to highlight the power of zebrafish as a model system for studying complex diseases which involve multiple genetic loci. We are interested in identifying and characterizing genes implicated in the blinding condition of glaucoma. Glaucoma is a complex disease that often involves multiple genetic loci. Most disease causing and modifying genes for glaucoma remain unidentified. However, several genes that regulate various aspects of ocular development have been shown to associate with glaucoma. With zebrafish, forward and reverse genetic approaches can be combined in order to identify critical genetic interactions required for normal and pathological events in the development and maintenance of the eye. PMID- 15533793 TI - Microarray gene expression profiling during the segmentation phase of zebrafish development. AB - We analyzed 15,512 unique transcripts from wild-type Danio rerio using a long oligonucleotide microarray containing >16,000 65-mers probes. Total RNA was isolated from staged embryos at 2 h intervals over a 24-h period. On average, at any given time point, 27% of the probe set detected corresponding transcripts in embryonic RNA. There were two predominant patterns in the nearly 4000 genes that changed expression in at least one time point during the first 24 hpf. At 12 hpf, we detected 420 up-regulated and 386 down-regulated genes. By 24 hpf, the number of up- and down-regulated genes had increased to 954 and 766, respectively. While the majority of these genes maintained their new level of expression for the duration of the time course, we identified five genes with phasic regulation over the 24-h time course. Two of these genes, germ cell nuclear factor and mesogenin, have been identified as being expressed during gastrulation (5 1/4 to 10 h postfertilization) and subsequently repressed. A cluster containing 36 distinct ribosomal proteins was up-regulated at 12 h, indicating a capability for de novo protein synthesis during and after this stage. Twenty-three muscle-specific genes were up-regulated late during the initial 24 hpf, corresponding to the development and differentiation of the somites. PMID- 15533794 TI - The effects of temperature reduction on gene expression and oxidative stress in skeletal muscle from adult zebrafish. AB - Longevity is inversely proportional to ambient temperature in ectothermic organisms such as fish. However, the mechanism by which reducing temperature over a physiological range increases life span is not known and available data are derived primarily from invertebrates. With a rodent-like longevity and abundant biological resources, the zebrafish is an ideal vertebrate ectothermic model in which to investigate this phenomenon. As an initial approach, the effects of a year-long 10 degrees C reduction in water temperature on global gene expression in tail skeletal muscle from adult zebrafish were determined using an oligonucleotide microarray representing 15,512 genes. Expression levels for approximately 600 genes were up-regulated by 1.7-fold or greater by the reduction in temperature, while a similar number of transcripts were down regulated by more than 1.7-fold. Using gene ontology (GO) classifications for molecular function, two functional groups, "oxygen and reactive oxygen species metabolism" and "response to oxidative stress," were found to be overrepresented among up regulated genes. Transcripts levels for the genes in these two categories were increased by temperature reduction (TR). However, temperature reduction did not suppress lipid peroxidation potential, protein carbonyl content, or 8-oxoguanine level. Additional studies will be required to further delineate the role of altered gene expression and oxidative stress on the longevity-promoting effects of temperature reduction. PMID- 15533795 TI - Aryl hydrocarbon receptor (AhR)-independent effects of 2,3,7,8-tetrachlorodibenzo p-dioxin (TCDD) on softshell clam (Mya arenaria) reproductive tissue. AB - A high prevalence of germinomas has been observed in certain populations of Mya arenaria from eastern Maine. The etiology of these tumors is unknown. We are investigating the hypothesis that exposure to environmental contaminants, such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) contributes to gonadal carcinogenesis. Clams were exposed to TCDD with or without the initiating compound diethylnitrosamine (DEN) in an attempt to induce germinomas. A TCDD-dependent alteration in gametogenesis was observed in which 32.5+/-6.5% of individuals exhibited undifferentiated gonads. Analyses of AhR and p53 expression were carried out to identify similarities between naturally occurring neoplastic and TCDD (+/-DEN)-altered reproductive tissues. Neoplastic tissues had significantly less p53 protein than matched controls, whereas TCDD-induced undifferentiated samples exhibited no difference in p53 protein levels compared to controls. No gender-specific differences were observed in AhR mRNA, but there were significant differences in protein levels. AhR was undetectable in male gonadal tissue whereas females exhibited a significant positive relationship between AhR protein levels and stage of ovogenesis. Despite exhibiting some morphological similarity, we conclude the TCDD-induced pathology is not a germinoma. We further suggest the change in reproductive tissue is due to inhibition of cell differentiation and/or development by an AhR-independent mechanism. PMID- 15533796 TI - Mycobacteriosis in zebrafish (Danio rerio) research facilities. AB - The Zebrafish International Resource Center was established to support the zebrafish research community, and includes a diagnostic service. One of the most common diseases that we have diagnosed is mycobacteriosis, which represented 18% of the diagnostic cases submitted from November 1999 to June 2003. We describe here the severity of the disease and associated pathological changes of 24 diagnostic cases from 14 laboratories. Identifications of the bacteria are provided for seven of these cases. For two cases in which culture of the organism was not successful, these identifications were based on ribosomal DNA (rDNA) sequence analysis obtained directly from infected tissues. Biochemical characteristics and rDNA sequence analysis from cultures are reported for the other isolates. Two severe outbreaks from different facilities on different continents were associated with an organism identified as Mycobacterium haemophilum based on rDNA sequence from tissues. Another severe outbreak was associated with an organism most closely related to Mycobacterium peregrinum. These species are recognized pathogens of humans, but this is the first report of them from fish. Bacteria identified as Mycobacterium chelonae or M. abscessus were recovered from fish in cases categorized as moderate disease or as an incidental finding. These findings indicate that species of Mycobacterium previously undescribed from fish (i.e., M. haemophilum and M. peregrinum) may pose significant health problems in zebrafish research facilities, whereas species and strains that are already recognized as common in fish usually cause limited disease on a population basis in zebrafish. PMID- 15533797 TI - In vitro response of the striped bass natural resistance-associated macrophage protein, Nramp, to LPS and Mycobacterium marinum exposure. AB - Mycobacteriosis in Chesapeake Bay (USA) striped bass Morone saxatilis is an ongoing disease problem with important economic implications for a large commercial and recreational fishery. Additionally, striped bass serve as a reservoir of potential mycobacterial zoonoses. Recently, we described a striped bass gene homolog of the natural resistance-associated macrophage protein family (MsNramp), which is responsible for resistance to mycobacterial infections in mice. Striped bass MsNramp is strongly induced in peritoneal exudate cells (PE) in vivo after intraperitoneal injection with Mycobacterium spp. The purpose of the present study was to investigate short-term in vitro MsNramp expression and reactive oxygen intermediate (ROI) production in primary cultures of adherent PE after exposure to bacterial lipopolysaccharide (LPS), or live- or heat-killed (HK) Mycobacterium marinum. PE expressed significantly higher levels of MsNramp at 4 and 24 h post-treatment with live and HK M. marinum. MsNramp response to LPS was dose-dependent in these cells, with maximum expression at 4 h and 20 microg/ml LPS. Treatment of PE with LPS resulted in increased intracellular superoxide anion levels, whereas treatment with live M. marinum caused a significant depression. This study is the first report of induction of a teleost Nramp in vitro by mycobacteria, and supports findings of teleost Nramp induction by LPS. PMID- 15533798 TI - Patterns of transcription of a virus-like agent in tumor and non-tumor tissues in bicolor damselfish. AB - Damselfish neurofibromatosis (DNF) is a transmissible disease characterized by peripheral nerve sheath and pigment cell tumors which occurs in bicolor damselfish (Stegastes partitus) on Florida reefs. The damselfish virus-like agent (DVLA) is associated with the development of DNF and contains a 2.4-kb DNA genome which was found at high levels in tumors and tumor-derived cell lines and at lower levels in non-tumor tissues of both spontaneously diseased fish (TF) and fish with experimentally induced tumors (EF). An analysis of transcription patterns revealed up to five DVLA derived RNAs ranging in size from 300 to 1400 bp in these cell types. DNA was the most commonly distributed DVLA component in TF and EF followed by RNA. Prevalence of transcripts varied by tissue type. The smallest transcripts were the most common in all cell types and the most complete patterns, which included the larger transcripts, were observed primarily in tumors. The presence of viral RNAs in addition to DNA in non-tumor tissues suggested these tissues were infected by DVLA and indicated a wide tissue tropism for this agent. The high levels of DVLA nucleic acids found in tumors suggest that replication is occurring there. However, the potential for DVLA replication in other tissues where only a limited range of transcripts were present is not known. The mechanism of tumorigenesis by this agent is unknown. However, the association of the larger transcripts with most tumor tissues and their absence in most non-tumor tissues suggests that these RNAs may be involved in tumor formation. PMID- 15533801 TI - Interstitial brachytherapy alone after breast conserving surgery: interim results of a German-Austrian multicenter phase II trial. AB - PURPOSE: To evaluate the value of interstitial brachytherapy (iBT) alone in the treatment of low-risk breast cancer patients with regard to local control, side effects, and cosmesis. METHODS AND MATERIALS: From November 2000 to January 2004, 176 patients with low-risk breast cancer were treated with iBT only. Patients were eligible for entering the study if: the tumor size was <3 cm; resection margins were clear by at least 2 mm; there were no lymph node metastases or only one micrometastasis (pNo, pNmi); age was >35 years; steroid hormone receptor was positive; and histologic grade was 1 or 2. Seventy-five percent of patients received pulsed-dose-rate brachytherapy (D(ref) = 50 Gy); 25% of patients received high-dose-rate brachytherapy (D(ref) = 32.0 Gy). An interim analysis is presented for all patients after an interim follow-up of 12 months, and for half the patient population with an interim follow-up of 21 months. RESULTS: All patients remained disease-free on the date of analysis. A perioperative complication breast infection was recorded for 1/176 (0.6%) patients. Late toxicity i.e., hypersensation, hyperpigmentation, fibrosis, or teleangiectasia was observed in 1-12% of all patients. Grade I Fibrosis was registered in 7.6% (13/172) and grade II in 7.0% (12/172) of evaluable patients. Similarly, grade I teleangiectasia was observed in 4.7% (8/172), grade II in 0.6% (1/172), and grade III also in 0.6% (1/172) of evaluable patients. Excellent or good cosmetic results have been observed in 92-95% of patients. CONCLUSIONS: Brachytherapy as monotherapy in low-risk breast cancer patients after breast-conserving surgery is an effective, precise treatment modality without perioperative morbidity, low acute, mild late toxicity, and yields good to excellent cosmetic results. PMID- 15533802 TI - Is simulation necessary for each high-dose-rate tandem and ovoid insertion in carcinoma of the cervix? AB - PURPOSE: To evaluate the dose variation in high-dose-rate (HDR) intracavitary brachytherapy for cancer of the cervix when treatment planning is performed prior to each applicator insertion versus when the initial plan is used for each treatment. METHODS AND MATERIALS: Fourteen patients with carcinoma of the cervix were treated with chemoradiotherapy followed by five intracavitary tandem and ovoid insertions of 600 cGy/fraction. We modified the actual plans to calculate the dose each dose point would have received using only the treatment plan created for the initial fraction. RESULTS: An increase in the percent dose to the rectum, bladder, and vaginal surface of 5%, cGy (p = 0.038), 6% (p = 0.006), and 11%, respectively, were observed when the initial treatment plan was used versus using the optimized treatment plan for each insertion. The greatest single change resulted in a percent increase of 35%, 30%, and 45% to the rectum, bladder, and vaginal surface points, respectively. CONCLUSIONS: Increased dose to at-risk structures occurred when individualized treatment planning was not performed. Since a significant increase in dose to the rectum (p = 0.038) and bladder (p = 0.006) was obtained without customized treatment planning, we continue to advocate individualized treatment planning in HDR tandem and ovoid insertions for the treatment of cervix cancer. PMID- 15533803 TI - Factors predicting an increased dose to the penile bulb in permanent seed prostate brachytherapy. AB - PURPOSE: Erectile dysfunction following permanent seed brachytherapy for prostate cancer may be related to the dose to the penile bulb. We investigated anatomic and dosimetric factors that might contribute to an increased dose to the penile bulb. METHODS AND MATERIALS: One-month CT and MR images were examined for 50 consecutive patients treated with exclusive (125)I permanent seed prostate brachytherapy to a prescribed dose of 145 Gy. Implants were preplanned by transrectal ultrasound (TRUS). Postimplant dosimetry was performed at 1 month using an MRI-CT fusion. Spearman's correlation was used to establish a correlation between dosimetric parameters, anatomical factors, and the dose to the penile bulb. RESULTS: Penile bulb volumes ranged from 1.2-8.5 cc (median, 3.9 cc). The distance from the penile bulb to the prostate apex ranged from 5-33 mm (median, 15.5 mm). D50 of the penile bulb ranged from 13-121 Gy. The range for the V45 (65 Gy) was 0-87%; only 3% of patients had >2 cc covered by this isodose and in 16% of patients the V45 covered more than 50% of the penile bulb. About one-third of the patients received a dose to the bulb that would put them at a high risk of erectile dysfunction after external beam radiation, if the dose were radiobiologically equivalent. There was a significant inverse correlation between the distance between the prostate apex and the penile bulb, and the dosimetric parameters of the bulb: r = -0.548, -0.656, p = < 0.01. The further caudal the apex was from the symphysis, the closer it was to the penile bulb (r = -0.564, p = <0.01). We could not find a correlation between the dose to the prostate or its apex and the dose to the penile bulb. CONCLUSION: When the prostate apex is close to the penile bulb, care should be taken to limit the dose to the penile bulb, if possible. This may reduce the incidence of erectile dysfunction and urinary toxicity after permanent seed prostate brachytherapy. PMID- 15533804 TI - Factors predictive of rectal bleeding after 103Pd and supplemental beam radiation for prostate cancer. AB - PURPOSE: To evaluate the contribution of various clinical and radiation treatment parameters to the likelihood of late rectal bleeding after brachytherapy plus supplemental beam radiation (EB). METHODS: A total of 161 intermediate risk patients, with Gleason score 7 or higher and/or PSA 10-20 ng/ml randomized to implantation with (103)Pd (90 versus 115 Gy) with 44 versus 20 Gy EB (2 Gy/day) were studied. Beam radiation was delivered with a four-field arrangement designed to cover the prostate and seminal vesicles with a 2 cm margin (reduced to 1.0 cm posteriorly). Isotope implantation was performed by standard techniques, using a modified peripheral loading pattern. A postimplant CT scan (3 mm slice thickness) was obtained 1-4 h after implantation. Dose volume histograms of the prostate and rectum were calculated using the outer prostatic and rectal margins identified on CT scan by one investigator (KW). Rectal doses were expressed as the R100, R200, and R300, defined as the rectal volume (cc) that received at least 100%, 200%, or 300% of the prescription dose, respectively. External beam doses were expressed as EB75% (cc)-the volume of rectum that received 75% of the beam prescription dose. Treatment-related rectal morbidity was monitored by mailed questionnaires, using Radiation Therapy Oncology Group (RTOG) criteria, at 1, 3, 6, 12, 24, and 36 months. Patients who reported Grade 1 or higher RTOG morbidity were contacted by telephone to obtain more details regarding their rectal bleeding. RESULTS: In univariate analysis, rectal bleeding was statistically related to the R100, R200, and R300 values, with p-values of 0.0055, 0.0007, and 0.012, respectively. Bleeding was not related to gap times, prostate size, patient age, V100 or D90 values. The EB75% values were similar in 44 Gy patients with or without late bleeding. CONCLUSION: Considering the potential severity of rectal morbidities and their relationship to implant dose, we urge our colleagues to routinely monitor the rectal implant doses of their own patients to make sure that such doses are kept within an accepted range. PMID- 15533805 TI - Transperineal TRUS-guided prostate brachytherapy using loose seeds versus RAPIDStrand: a dosimetric analysis. AB - PURPOSE: An analysis of the effect of stranded (125)I and loose (predominantly (103)Pd) sources on dosimetric outcomes of brachytherapy of the prostate. METHODS AND MATERIALS: Between September 1998 and December 2003, 473 patients were treated with brachytherapy for biopsy-proven carcinoma of the prostate. Of these, 337 (71%) procedures were performed using free seeds placed with a Mick applicator. Beginning in April 2002, a program of stranded (125)I sources (RAPIDStrand) was implemented; 136 (29%) patients were treated via this approach. Dosimetric variables were collected, as were events of urinary retention. RESULTS: Mean V100 values for the stranded (125)I approach were greater than those for free seeds (p < 0.0005), whether (125)I or (103)Pd (p < 0.005). Use of the strand was the most significant determinant of V100 of all variables examined. The stranded (125)I approach was also associated with higher mean D90 values and lower V150-urethral doses. CONCLUSIONS: Use of stranded (125)I was associated with superior dosimetric outcomes in this group of patients. PMID- 15533806 TI - Temporal effect of neoadjuvant androgen deprivation therapy on PSA kinetics following permanent prostate brachytherapy with or without supplemental external beam radiation. AB - PURPOSE: To evaluate the effect of neoadjuvant androgen deprivation therapy on PSA kinetics following brachytherapy with or without supplemental external beam radiation therapy (XRT), to evaluate the magnitude and duration of the peak PSA, and to compare potential differences in PSA response curves between biochemically disease-free and failed patients. MATERIALS AND METHODS: From November 1995 through August 2000, 179 consecutive patients with clinical T1b-T3a NxM0 (2002 AJCC) prostate cancer received neoadjuvant androgen deprivation therapy (median, 4 months) prior to brachytherapy using (103)Pd or (125)I with or without supplemental XRT. The median follow-up was 56 months. Following brachytherapy, PSA determinations were obtained at 3 months and then every 6 months thereafter. A median and mean of 9 and 9.5 PSA determinations, respectively, were obtained per patient. Biochemical disease-free survival was defined by the ASTRO and Houston definitions with both definitions used to determine the rate of overestimation of early biochemical failure secondary to testosterone recovery. RESULTS: Although a trend for higher baseline and peak PSA was noted in (125)I monotherapy patients, the difference between the 4 groups was not statistically significant (p = 0.088). Changes in PSA over time, however, were statistically significant (p = 0.042). For all 4 groups, the peak PSA occurred approximately 15 21 months following brachytherapy. For biochemically disease-free patients, the median PSA increase above nadir was 0.1 ng/mL, and the median number of consecutive rises in PSA was 1. Using the ASTRO and Houston definitions, only 2.3% and 0.6% of patients would have been inadvertently scored as failures. CONCLUSIONS: In brachytherapy patients receiving 3-6 months of neoadjuvant androgen deprivation therapy, the median PSA increase above nadir was 0.1 ng/mL. The ASTRO and Houston definitions of biochemical failure inadvertently scored only 2.3% and 0.6% of patients as failures. PMID- 15533807 TI - Comparison of inverse planning simulated annealing and geometrical optimization for prostate high-dose-rate brachytherapy. AB - PURPOSE: An inverse planning simulated annealing (IPSA) algorithm for optimization of high-dose-rate (HDR) brachytherapy has been previously described. In this study, IPSA is compared with geometrical optimization (GO) for prostate brachytherapy. METHODS AND MATERIALS: Using CT data collected from 10 patients, treatment plans were prepared using GO and IPSA. The clinical target volume (CTV) and critical organs (CO) including bladder, rectum, and urethra were contoured using Plato Version 14.2.1 (Nucletron Corp., Veenendaal, The Netherlands). Implant catheters were digitized using the CT planning system. All dwell positions outside of the CTV were turned off. Two optimized plans were generated for each implant using GO and IPSA. The same set of dose constraints were used for all inverse planning calculations and no manual adjustment of the dwell weight was used. Two prescription methods were used. Using the first method, coverage was prioritized: the prescription dose was normalized to the isodose volume that covers 98% of the CTV (V100 = 98% of CTV). The dose volume histograms (DVH) of CO were generated for comparison. Using the second method, sparing was prioritized: the prescription dose was normalized such that no urethra volume received 150% of the prescription dose (V150-urethra = 0 cc). The DVH of CTV and CO were generated, and the homogeneity index (HI) and conformal index (COIN) were calculated for comparison and compared using the Wilcoxon matched-pairs test. RESULTS: Using the coverage-prioritized method, the difference in V80-bladder dose was not statistically significant (p = 0.09; median: IPSA = 0.62 cc, GO = 1.05 cc). The V80-rectum ranged from 0.20-4.8 cc, and 0.05-1.4 cc using GO and IPSA, respectively. IPSA's V80-rectum was significantly lower (p = 0.005; median: IPSA=0.38 cc, GO = 1.31 cc). V150-urethra ranged from 0.02-0.75 cc and 0.0-0.01 cc using GO and IPSA, respectively. The V150-urethra was significantly lower using IPSA (p = 0.005; median: IPSA = 0.00 cc, GO = 0.33 cc). Using the sparing prioritized method, the V100-prostate ranged from 30-97% and 95-100% using GO and IPSA, respectively. This difference was statistically significant (p = 0.008). The HI and COIN were statistically higher using IPSA (p = 0.005). CONCLUSION: Anatomy-based inverse optimization using IPSA is superior to dwell-position-based optimization using GO as it: (1) Improves target coverage and conformality while sparing normal structures, (2) Improves dose homogeneity within the target, and (3) Minimizes volume of non-contoured normal tissue irradiated. Routine application of three-dimensional brachytherapy planning and anatomy-based inverse dwell time optimization is recommended. PMID- 15533808 TI - Chronic pelvic pain following prostate brachytherapy: a case report. AB - PURPOSE: To alert physicians and potential patients that chronic postimplant pelvic pain syndromes can occur, and that dosimetric parameters (i.e., implant technique) may predispose patients to it. METHODS AND MATERIALS: The authors are currently following 3 prostate brachytherapy patients with what appear to be chronic radiation-related pelvic pain, variously exacerbated by urination or perineal pressure. The 3 patients were identified in the course of routine follow up, and do not represent a concerted attempt to identify such patients from a larger group of patients being followed by the authors. Three control groups of 10 patients each treated with (125)I, (103)Pd, or (103)Pd + external beam radiation and with no reported dysuria at 6 months postimplant were taken from two ongoing prospective trials. The 3 patients reported here were each administered a brief questionnaire regarding the effect of their urinary pain on daily activities. RESULTS: Patients with chronic pain tended to have high central prostatic doses, at least on some planes. Maximal, mean, and median urethral doses were higher for patients with chronic pain, but there was some overlap with control patients. The prostate V100s were similar between patients with chronic pain and controls, but there was a trend toward higher V200s and V300s in pain syndrome patients. CONCLUSION: Recalcitrant brachytherapy-related pelvic pain is an uncommon occurrence that may be partly related to higher central prostatic doses. PMID- 15533809 TI - Development of a rat solid tumor model for continuous low-dose-rate irradiation studies using 125I and 103Pd sources. AB - PURPOSE: To develop an experimental technique for studying the radiobiology of continuous low-dose-rate irradiation (CLDRI) using clinical brachytherapy sources emitting low energy photons for a rat solid tumor model. METHODS AND MATERIALS: BA1112 tumors were grown between the ears of 14-week-old male WAG/Rij rats by interdermal inoculation. A radioactive source afterloading system, which consists of a lightweight helmet sutured to the rat and a nine-source polystyrene applicator, was fabricated for in vivo tumor irradiation by (125)I and (103)Pd brachytherapy sources. This system has a 12 x 12 mm opening in the center to accommodate the tumor and its growth during irradiation (the diameter of a typical BA1112 tumor was about 6 mm when radiation was applied). The spatial locations of the nine sources were optimized to produce an as uniform as possible three-dimensional dose distribution to the central portion of the applicator for both the (125)I and (103)Pd sources. Absolute dose delivered by the applicator was verified by point dose measurements using calibrated TLD in a polystyrene phantom that mimics the scattering environment of the tumor on the rat. RESULTS: The feasibility of tumor cure experiments using the experimental technique presented in this work was demonstrated. The technique was used to study the influence of initial dose rate on the in vivo tumor cure probability of BA1112 tumors irradiated by (125)I and (103)Pd sources at dose rates varying from 8-20 cGy/h. The technique was also used for studying the in vitro tumor cell survival following in vivo CLDRI irradiation of the tumor. CONCLUSION: An experimental technique using an in vivo tumor model has been developed for studying the radiobiological effects of continuous low-dose-rate irradiations using (125)I sources alone, (103)Pd sources alone, or a mixture of (125)I and (103)Pd sources. PMID- 15533810 TI - Intravascular brachytherapy using 90Sr for saphenous vein grafts having diameters ranging from 2.0-5.0 mm. AB - PURPOSE: Symptomatic coronary artery disease is routinely treated with angioplasty and stenting. Unfortunately, treatment failure in the form of in stent restenosis (ISR) occurs relatively frequently. Intravascular brachytherapy (IVBT) is a safe and effective method proven to markedly reduce the rate of ISR in native coronary arteries. The commercially available devices for IVBT are not FDA-approved for treatment of saphenous vein grafts (SVG). This article presents calculated dosimetry for treatment of a wide range of SVG, in addition to further evaluating the dose homogeneity for native coronary arteries. METHODS AND MATERIALS: AAPM Task Group 43 and 60 formalisms permitted dose calculations for a wide range of vessel internal diameters (phi) in both native coronary arteries and SVG. Doses were analytically calculated for the Novoste Beta-Cath 5.0 French (F) treatment devices (30, 40, and 60 mm sourcetrains) when employed for the treatment of native vessels with 2.7 T and G --> C transversion mutations and polymerase arrest. These findings suggested that further oxidized lesions of 8-oxoG are more mutagenic and toxic than 8-oxoG itself. Repair of oxidatively damaged bases, including Sp and Gh/Ia, are initiated by the base excision repair (BER) system that involves the DNA glycosylases Fpg, Nei, and Nth in E. coli. Mammalian homologs of two of these BER enzymes, OGG1 and NTH1, have little or no affinity for Gh/Ia and Sp. Herein we report that two recently identified mammalian glycosylases, NEIL1 and NEIL2, showed a high affinity for recognition and cleavage of DNA containing Gh/Ia and Sp lesions. NEIL1 and NEIL2 recognized both of these lesions in single-stranded DNA and catalyzed the removal of the lesions through a beta- and delta elimination mechanism. NEIL1 and NEIL2 also recognized and excised the Gh/Ia lesion opposite all four natural bases in double-stranded DNA. NEIL1 was able to excise the Sp lesion opposite the four natural bases in double-stranded DNA, however, NEIL2 showed little cleavage activity against the Sp lesion in duplex DNA although DNA trapping studies show recognition and binding of NEIL2 to this lesion. This work suggests that NEIL1 and NEIL2 are essential in the recognition of further oxidized lesions arising from 8-oxoG and implies that these BER glycosylases may play an important role in the repair of DNA damage induced by carcinogenic metals. PMID- 15533837 TI - A role for Pms2 in the prevention of tandem CC --> TT substitutions induced by ultraviolet radiation and oxidative stress. AB - DNA mismatch repair (MMR) is important for preventing base-pair substitutions caused by spontaneous or damage-related DNA polymerase errors. We have used a reversion assay based on mouse Aprt to investigate the role of MMR in preventing ultraviolet radiation (UV) and oxidative stress induced tandem CC --> TT base pair substitutions in cultured mammalian cells. The reversion construct used for this assay can detect both C --> T and CC --> TT mutational events. Most spontaneous mutations in Pms2-deficient cells were single C --> T substitutions (88%), with the remainder being tandem CC --> TT substitutions (12%). The percentage of tandem CC --> TT substitutions rose to 64% and 94% for Pms2 deficient cells exposed to UV and a mixture of hydrogen peroxide and metals (Cu/Fe), respectively. Exposure to hydrogen peroxide alone or metals alone did not induce the tandem substitutions, nor did treatment of the cells with the alkylating agent ethylmethane sulfonate, which induces G --> A substitutions on the opposite strand. Tandem CC --> TT substitutions were also induced by UV irradiation and the hydrogen peroxide/metal mixture in Pms2-proficient cells, but at frequencies significantly lower than those observed in the Pms2-deficient cells. We conclude that mismatch repair plays an important role in preventing tandem CC --> TT substitutions induced by certain genotoxin exposures. PMID- 15533838 TI - The Saccharomyces cerevisiae PDS1 and RAD9 checkpoint genes control different DNA double-strand break repair pathways. AB - In response to DNA damage, the Saccharomyces cerevisiae securin Pds1 blocks anaphase promotion by inhibiting ESP1-dependent degradation of cohesins. PDS1 is positioned downstream of the MEC1- and RAD9-mediated DNA damage-induced signal transduction pathways. Because cohesins participate in postreplicative repair and the pds1 mutant is radiation sensitive, we identified DNA repair pathways that are PDS1-dependent. We compared the radiation sensitivities and recombination phenotypes of pds1, rad9, rad51 single and double mutants, and found that whereas pds1 rad9 double mutants were synergistically more radiation sensitive than single mutants, pds1 rad51 mutants were not. To determine the role of PDS1 in recombinational repair pathways, we measured spontaneous and DNA damage associated sister chromatid exchanges (SCEs) after exposure to X rays, UV and methyl methanesulfonate (MMS) and after the initiation of an HO endonuclease generated double-strand break (DSB). The rates of spontaneous SCE and frequencies of DNA damage-associated SCE were similar in wild type and pds1 strains, but the latter exhibited reduced viability after exposure to DNA damaging agents. To determine whether pds1 mutants were defective in other pathways for DSB repair, we measured both single-strand annealing (SSA) and non-homologous end joining (NHEJ) in pds1 mutants. We found that the pds1 mutant was defective in SSA but efficient at ligating cohesive ends present on a linear plasmid. We therefore suggest that checkpoint genes control different pathways for DSB repair, and PDS1 and RAD9 have different roles in recombinational repair. PMID- 15533840 TI - Mismatch repair protein Msh2 contributes to UVB-induced cell cycle arrest in epidermal and cultured mouse keratinocytes. AB - Nucleotide excision repair (NER), cell cycle regulation and apoptosis are major defence mechanisms against the carcinogenic effects of UVB radiation. NER eliminates UVB-induced DNA photolesions via two subpathways: global genome repair (GGR) and transcription-coupled repair (TCR). In a previous study, we found UVB induced accumulation of tetraploid (4N) keratinocytes in the epidermis of Xpc(-/ ) mice (no GGR), but not in Xpa(-/-) (no TCR and no GGR) or in wild-type (WT) mice. We inferred that this arrest in Xpc(-/-) mice is caused by erroneous replication past photolesions, leading to 'compound lesions' known to be recognised by mismatch repair (MMR). MMR-induced futile cycles of breakage and resynthesis at sites of compound lesions may then sustain a cell cycle arrest. The present experiments with Xpc(-/-)Msh2(-/-) mice and derived keratinocytes show that the MMR protein Msh2 indeed plays a role in the generation of the UVB induced arrested cells: a Msh2-deficiency lowered significantly the percentage of arrested cells in vivo (40-50%) and in vitro (30-40%). Analysis of calyculin A (CA)-induced premature chromosome condensation (PCC) of cultured Xpc(-/-) keratinocytes showed that the delayed arrest occurred in late S phase rather than in G(2)-phase. Taken together, the results indicate that in mouse epidermis and cultured keratinocytes, the MMR protein Msh2 plays a role in the UVB-induced S phase arrest. This indicates that MMR plays a role in the UVB-induced S-phase arrest. Alternatively, Msh2 may have a more direct signalling function. PMID- 15533839 TI - DNA glycosylase activities for thymine residues oxidized in the methyl group are functions of the hNEIL1 and hNTH1 enzymes in human cells. AB - Bacteria and eukaryotes possess redundant activities that recognize and remove oxidatively damaged bases from DNA through base excision repair. DNA glycosylases excise damaged bases to initiate the base excision repair pathway. hOgg1 and hNTH1, homologues of E. coli MutM and Nth, respectively, had been identified and characterized in human cells. Recent works revealed that human cells have three orthologues of E. coli Nei, hNEIL1, hNEIL2 and hNEIL3. In the present experiments, hNEIL1 protected the E. coli nth nei mutant from lethal effect of hydrogen peroxide and high frequency of spontaneous mutations under aerobic conditions. Furthermore, hNEIL1 efficiently cleaved double stranded oligonucleotides containing 5-formyluracil (5-foU) and 5-hydroxymethyluracil (5 hmU) in vitro via beta- and delta-elimination reactions. Similar activities were detected with hNTH1. These results indicate that hNEIL1 and hNTH1 are DNA glycosylases that excise 5-foU and 5-hmU as efficiently as Tg in human cells. PMID- 15533841 TI - DNA damage recognition and repair by the murine MutY homologue. AB - E. coli MutY excises adenine from duplex DNA when it is mispaired with the mutagenic oxidative lesion 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG). While E. coli MutY has been extensively studied, a detailed kinetic analysis of a mammalian MutY homologue has been inhibited by poor overexpression in bacterial hosts. This current work is the first detailed study of substrate recognition and repair of mismatched DNA by a mammalian adenine glycosylase, the murine MutY homologue (mMYH). Similar to E. coli MutY, the processing of OG:A substrates by mMYH is biphasic, indicating that product release is rate-limiting. Surprisingly, the intrinsic rates of adenine removal from both OG:A and G:A substrates by mMYH are diminished ( approximately 10-fold) compared to E. coli MutY. However, similar to E. coli MutY, the rate of adenine removal is approximately nine-fold faster with an OG:A- than a G:A-containing substrate. Interestingly, the rate of removal of 2-hydroxyadenine mispaired with OG or G in duplex DNA by mMYH was similar to the rate of adenine removal from the analogous context. In contrast, 2 hydroxyadenine removal by E. coli MutY was significantly reduced compared to adenine removal opposite both OG and G. Furthermore, dissociation constant measurements with duplexes containing noncleavable 2'-deoxyadenosine analogues indicate that mMYH is less sensitive to the structure of the base mispaired with OG or G than MutY. Though in many respects the catalytic behavior of mMYH is similar to E. coli MutY, the subtle differences may translate into differences in their in vivo functions. PMID- 15533842 TI - Genetic studies of recombining DNA in pneumococcal transformation. 1966. PMID- 15533843 TI - Methyl-directed repair of DNA base-pair mismatches in vitro. 1983. PMID- 15533844 TI - Intrapleural instillation of rituximab for the treatment of malignant pleural effusions in NHL. AB - A patient with CD20(+) leukaemic lymphoplasmacytic Non-Hodgkin's lymphoma (NHL) presented with bilateral malignant pleural effusions. Systemic chemotherapy, repeated percutaneous drainage and bilateral continous chest tube drainage were unable to control the effusions. Rituximab was instilled in a dose-escalating manner via the chest tubes into both pleural spaces, within two weeks the effusions resolved, and the patient has stayed free of symptoms for eight months ongoing. Rituximab may be a promising novel treatment option for malignant effusions in CD20(+) NHL. PMID- 15533845 TI - Crystalline inclusions in B monoclonal lymphocytes. PMID- 15533846 TI - Clinical and molecular complete remission in a case of variant hairy cell leukemia treated with DHAP followed by high-dose chemotherapy plus rituximab. PMID- 15533847 TI - Breakthrough zygomycosis during voriconazole treatment for invasive aspergillosis. PMID- 15533848 TI - Evaluation of gemcitabine in relapsed or refractory multiple myeloma. PMID- 15533849 TI - The apparent excess of acute promyelocytic leukemia in infant acute leukemias in Brazil. PMID- 15533850 TI - Hepatitis B and C viruses and Hodgkin lymphoma: a case-control study from Northern and Southern Italy. PMID- 15533851 TI - Combination of isosorbide dinitrate and hydralazine in blacks with heart failure. AB - BACKGROUND: We examined whether a fixed dose of both isosorbide dinitrate and hydralazine provides additional benefit in blacks with advanced heart failure, a subgroup previously noted to have a favorable response to this therapy. METHODS: A total of 1050 black patients who had New York Heart Association class III or IV heart failure with dilated ventricles were randomly assigned to receive a fixed dose of isosorbide dinitrate plus hydralazine or placebo in addition to standard therapy for heart failure. The primary end point was a composite score made up of weighted values for death from any cause, a first hospitalization for heart failure, and change in the quality of life. RESULTS: The study was terminated early owing to a significantly higher mortality rate in the placebo group than in the group given isosorbide dinitrate plus hydralazine (10.2 percent vs. 6.2 percent, P=0.02). The mean primary composite score was significantly better in the group given isosorbide dinitrate plus hydralazine than in the placebo group ( 0.1+/-1.9 vs. -0.5+/-2.0, P=0.01; range of possible values, -6 to +2), as were its individual components (43 percent reduction in the rate of death from any cause [hazard ratio, 0.57; P=0.01] 33 percent relative reduction in the rate of first hospitalization for heart failure [16.4 percent vs. 22.4 percent, P=0.001], and an improvement in the quality of life [change in score, -5.6+/-20.6 vs. 2.7+/-21.2, with lower scores indicating better quality of life; P=0.02; range of possible values, 0 to 105]). CONCLUSIONS: The addition of a fixed dose of isosorbide dinitrate plus hydralazine to standard therapy for heart failure including neurohormonal blockers is efficacious and increases survival among black patients with advanced heart failure. PMID- 15533852 TI - Race-based therapeutics. PMID- 15533853 TI - Nitroso-redox balance in the cardiovascular system. PMID- 15533854 TI - Incidence of hepatitis C virus and HIV among new injecting drug users in London: prospective cohort study. PMID- 15533855 TI - Placebo-controlled, randomized clinical trial of azimilide for prevention of ventricular tachyarrhythmias in patients with an implantable cardioverter defibrillator. AB - BACKGROUND: Although implanted cardioverter defibrillators (ICDs) effectively treat sustained ventricular tachyarrhythmias, up to 50% of ICD recipients eventually require concomitant antiarrhythmic drug therapy to prevent symptomatic arrhythmia recurrences and hence reduce the number of device therapies. METHODS AND RESULTS: A total of 633 ICD recipients were enrolled in a randomized, double blind, placebo-controlled study to evaluate the effect of daily doses of 75 or 125 mg of azimilide on recurrent symptomatic ventricular tachyarrhythmias and ICD therapies. Total all-cause shocks plus symptomatic ventricular tachycardia (VT) terminated by antitachycardia pacing (ATP) were significantly reduced by azimilide, with relative risk reductions of 57% (hazard ratio [HR]=0.43, 95% CI 0.26 to 0.69, P=0.0006) and 47% (HR=0.53, 95% CI 0.34 to 0.83, P=0.0053) at 75- and 125-mg doses, respectively. The reductions in all-cause shocks with both doses of azimilide did not achieve statistical significance. The incidence of all appropriate ICD therapies (shocks or ATP-terminated VT) was reduced significantly among patients taking 75 mg of azimilide (HR=0.52, 95% CI 0.30 to 0.89, P=0.017) and those taking 125 mg of azimilide (HR=0.38, 95% CI 0.22 to 0.65, P=0.0004). Five patients in the azimilide groups and 1 patient in the placebo group had torsade de pointes; all were successfully treated by the device. One patient taking 75 mg of azimilide had severe but reversible neutropenia. CONCLUSIONS: Azimilide significantly reduced the recurrence of VT or ventricular fibrillation terminated by shocks or ATP in ICD patients, thereby reducing the burden of symptomatic ventricular tachyarrhythmia. PMID- 15533856 TI - Restitution properties during ventricular fibrillation in the in situ swine heart. AB - BACKGROUND: Although restitution has been hypothesized to determine action potential duration (APD) during ventricular fibrillation (VF), cardiac memory may also be important. METHODS AND RESULTS: Transmembrane recordings were made with a floating microelectrode from the anterior right ventricular wall in 6 pigs during up to 60 seconds of VF. The recordings were divided into 5-second intervals, and APD60 and the diastolic interval (DI) were calculated for each activation cycle throughout each interval. Stepwise linear regression was used to determine how well each APD60 [APD60(n)] was predicted by the 4 previous DIs (n-1, n-2, n-3, n 4) and the 3 previous APD60s (n-1, n-2, n-3). A mean+/-SD of 3+/-1.5 of the variables entered the regression equation. DI(n-1) (70% of intervals) and APD60(n 1) (71% of intervals) appeared most frequently in the regression equations and were the first or second variables entered during the stepwise regression in 87% and 76% of the intervals in which they were present, respectively. The coefficients of DI(n-1) and APD60(n-1) were positive 89% and 98% of the time, respectively. R2 of the regression for all entered variables during all intervals was 0.39+/-0.05. CONCLUSIONS: The high incidence and positive coefficient of DI(n 1) indicate that restitution is important in determining APD during VF, whereas the similarly high incidence and positive coefficient of APD(n-1) indicate that cardiac memory is equally important. The finding that the regression equation accounts for only 39% of the variability of APD indicates that factors other than restitution and memory are also important in determining APD during VF. PMID- 15533857 TI - Left atrial tachycardia originating from the mitral annulus-aorta junction. AB - BACKGROUND: At the mitral annulus-aorta (MA-Ao) junction, the left atrium is continuous through the subaortic curtain with the musculature of the anterior mitral leaflet. Under experimental conditions, this region can generate abnormal electrical activity. In patients with left atrial tachycardia, we investigated whether this region could be the source of this arrhythmia. METHODS AND RESULTS: In 10 (28%) of 35 consecutive patients with left atrial tachycardia, the arrhythmia originated from the MA-Ao junction. Sustained, self-limited episodes of atrial tachycardia (cycle length, 340+/-56 ms; duration, 125+/-69 seconds) were repeatedly induced. Prematurity of the extrastimulus and time to first atrial tachycardia complex were directly correlated (R=0.66; P<0.001). During tachycardia, bipolar electrograms at the earliest site preceded onset of the P wave by 44+/-14 ms and were of longer duration and lower amplitude than those recorded from nearby left atrial sites (52+/-8 versus 24+/-4 ms, P<0.001; and 0.53+/-0.08 versus 3.45+/-0.96 mV, respectively; P<0.001). Ablation eliminated the tachycardia with no recurrence after a mean follow-up of 24+/-19 months. A comparative study in mouse embryos demonstrated the presence of the developing specialized conduction system in the MA-Ao region starting at embryonic age 11.5. CONCLUSIONS: The MA-Ao junction can be a frequent source of left atrial tachycardia. This previously unrecognized site of origin may explain why catheter ablation has been less successful in eliminating left versus right atrial tachycardias. Remnants of the developing specialized conduction system could be the underlying substrate of this arrhythmia. PMID- 15533858 TI - Upregulation of myocardial estrogen receptors in human aortic stenosis. AB - BACKGROUND: Estrogen receptor (ER)-mediated effects have been associated with the modulation of myocardial hypertrophy in animal models and in humans, but ER expression in the human heart and its relation to hypertrophy-mediated gene expression have not yet been analyzed. We therefore investigated sex- and disease dependent alterations of myocardial ER expression in human aortic stenosis together with the expression of hypertrophy-related genes. METHODS AND RESULTS: ER-alpha and -beta, calcineurin A-beta, and brain natriuretic peptide (BNP) mRNA were quantified by real-time polymerase chain reaction in left ventricular biopsies from patients with aortic valve stenosis (n=14) and control hearts with normal systolic function (n=17). ER protein was quantified by immunoblotting and visualized by immunofluorescence confocal microscopy. ER-alpha mRNA and protein were increased 2.6-fold (P=0.003) and 1.7-fold (P=0.026), respectively, in patients with aortic valve stenosis. Left ventricular ER-beta mRNA was increased 2.6-fold in patients with aortic valve stenosis (P<0.0001). ER-alpha and -beta were found in the cytoplasm and nuclei of human hearts. A strong inverse correlation exists between ER-beta and calcineurin A-beta mRNA in patients with aortic valve stenosis (r=-0.83, P=0.002) but not between ER-alpha or -beta and BNP mRNA. CONCLUSIONS: ER-alpha and -beta in the human heart are upregulated by myocardial pressure load. PMID- 15533859 TI - Mobilization of CD34/CXCR4+, CD34/CD117+, c-met+ stem cells, and mononuclear cells expressing early cardiac, muscle, and endothelial markers into peripheral blood in patients with acute myocardial infarction. AB - BACKGROUND: Adult stem cells can contribute to myocardial regeneration after ischemic injury. Bone marrow and skeletal muscles contain a population of CXCR4+ cells expressing genes specific for muscle progenitor cells that can be mobilized into the peripheral blood. The aims of the study were (1) to confirm the presence of early tissue-committed cells expressing cardiac, muscle, and endothelial markers in populations of mononuclear cells in peripheral blood and (2) to assess the dynamics and magnitude of the mobilization of CD34+, CD117+, CXCR4+, c-met+, CD34/CD117+, and CD34/CXCR4+ stem cells into peripheral blood in relation to inflammatory and hematopoietic cytokines in patients with ST-segment-elevation acute myocardial infarction (STEMI). METHODS AND RESULTS: Fifty-six patients with STEMI (<12 hours), 39 with stable angina, and 20 healthy control subjects were enrolled. Real-time reverse transcription-polymerase chain reaction (RT-PCR) was used for detection of tissue-specific markers. The number of the cells was assessed by use of a flow cytometer on admission, after 24 hours, and after 7 days. RT-PCR revealed increased expression of mRNA (up to 3.5-fold increase) for specific cardiac (GATA4, MEF2C, Nkx2.5/Csx), muscle (Myf5, Myogenin, MyoD), and endothelial (VE-cadherin, von Willebrand factor) markers in peripheral blood mononuclear cells. The number of CD34/CXCR4+ and CD34/CD117+ and c-met+ stem cells in peripheral blood was significantly higher in STEMI patients than in stable angina and healthy subjects, peaking on admission, without further significant increase after 24 hours and 7 days. CONCLUSIONS: The study demonstrates in the setting of STEMI a marked mobilization of mononuclear cells expressing specific cardiac, muscle, and endothelial markers as well as CD34/CXCR4+ and CD34/CD117+ and c-met+ stem cells and shows that stromal cell derived factor-1 is an important factor influencing the mobilization. PMID- 15533860 TI - Assessment of systemic right ventricular function in patients with transposition of the great arteries using the myocardial performance index: comparison with cardiac magnetic resonance imaging. AB - BACKGROUND: Assessment of systemic right ventricular (RV) function is a key point in the follow-up of patients with transposition of the great arteries (TGA). Current echocardiographic assessment of RV function is at best an estimate, and cardiac magnetic resonance (CMR) is considered the gold standard. However, this technique is expensive, has limited availability, and requires significant expertise to acquire and interpret the images. The myocardial performance index (MPI) has recently been studied for assessment of pulmonary RV function and shows promise as a simple yet powerful tool for assessing patients with RV dysfunction of various origins. We set out to compare MPI and CMR assessment of systemic RV function in patients with TGA. METHODS AND RESULTS: Data from patients with TGA (11 with congenitally corrected TGA, 18 with surgically corrected TGA) who had CMR within 6 months of their echocardiogram were reviewed. The average systemic RV ejection fraction (RVEF) by CMR was 39.4+/-11.4%, and the systemic RVMPI for this group was 0.56+/-0.21. There was a strong negative correlation between the systemic RVMPI and systemic RVEF by CMR (r=-0.82, P<0.01). The systemic RVEF can be estimated from this formula: RVEF=65%-(45.2xMPI). CONCLUSIONS: MPI can be used in patients with systemic RVs to assess global function and to estimate an EF with good accuracy. PMID- 15533861 TI - Sympathetic nerve activity in response to hypotensive stress in the postural tachycardia syndrome. AB - BACKGROUND: Increased central sympathetic activity and/or deficient peripheral sympathetic nerve function are among the proposed pathophysiological changes in patients with the postural tachycardia syndrome (POTS). Little is known about sympathetic nerve outflow and its role in hemodynamic control in this disorder. METHODS AND RESULTS: We recorded peroneal muscle sympathetic nerve activity in 9 POTS patients and 9 control subjects at rest and during a nitroprusside-induced hypotensive stimulus. Baseline blood pressure (BP) and heart rate were significantly higher in POTS patients than in controls. At rest, the burst frequency was similar in POTS patients and controls (18.1+/-6.2 and 20.1+/-7.9 bursts/min, respectively; P=NS), whereas the burst incidence was significantly lower (23.1+/-6.8 versus 32.2+/-11.4 bursts/100 heartbeats, P<0.05). Nitroprusside increased sympathetic outflow significantly more in POTS patients than in controls despite a similar BP decrease (burst frequency 20.4+/-7.5 versus 12.1+/-4.1 bursts/min, P=0.008, and burst incidence 21.8+/-8.4 versus 14.4+/-5.2 bursts/100 heartbeats, P=0.03). The change in mean burst area, a measure of the number of actively firing sympathetic neurons, was similar in patients and controls (117+/-15% versus 114+/-21%, P=NS). CONCLUSIONS: At rest, the tachycardia and normal burst frequency result in normal or even elevated BP in POTS patients. During a hypotensive stimulus, cardiovascular homeostasis is maintained by the increased sympathetic outflow and normal heart rate response despite the lack of concomitant increase in mean burst area that is most likely due to sympathetic denervation. PMID- 15533862 TI - Evaluation of venous and arterial conduit patency by 16-slice spiral computed tomography. AB - BACKGROUND: Computed tomography has been shown to be useful in the evaluation of aortocoronary bypass grafts (CABG). This is the first prospective study to evaluate the accuracy of a new-generation scanner in the detection of patency and significant stenoses (>50% decrease in diameter) of venous and arterial grafts in patients with previous CABG. METHODS AND RESULTS: In 96 patients (80 males, mean age 62 years) with previous CABG, a multislice computed tomography (MSCT) scan was performed (collimation 16x0.625 mm). Patients with atrial fibrillation, renal failure, severe respiratory disease, severe heart failure, heart rate >70 bpm despite therapy, or unstable angina were excluded. A total of 285 conduits implanted on the native coronary arteries at the time of CABG were evaluated. MSCT data were analyzed by 2 independent radiologists and compared with the results of conventional angiography. Three patients were excluded from analysis. All conduits were judged evaluable in 84 patients. Among these patients, MSCT correctly diagnosed 54 occluded grafts and 4 significant stenoses on the body of the grafts. Of the 17 significant anastomotic lesions, MSCT correctly diagnosed 15. For these 84 patients, diagnostic accuracy was 99%, sensitivity was 97%, and specificity was 100%. When all 93 patients were considered, the sensitivity of MSCT in diagnosing significant stenoses was 96%. CONCLUSIONS: MSCT with the new generation scanner allows for accurate assessment of venous and arterial conduits in patients with previous CABG with a high degree of sensitivity and specificity. Exclusion criteria and radiation exposure remain limitations of the method. PMID- 15533863 TI - Excessive tumor necrosis factor activation after infarction contributes to susceptibility of myocardial rupture and left ventricular dysfunction. AB - BACKGROUND: We investigated the potential contributions of tumor necrosis factor alpha (TNF-alpha) on the incidence of acute myocardial rupture and subsequent chronic cardiac dysfunction after myocardial infarction (MI) in TNF knockout (TNF /-) mice compared with C57/BL wild-type (WT) mice. METHODS AND RESULTS: Animals were randomized to left anterior descending ligation or sham operation and killed on days 3, 7, 14, and 28. We monitored cardiac rupture rate, cardiac function, inflammatory response, collagen degradation, and net collagen formation. We found the following: (1) within 1 week after MI, 53.3% (n=120) of WT mice died of cardiac rupture, in contrast to 2.5% (n=80) of TNF-/- mice; (2) inflammatory cell infiltration and cytokine expression were significantly higher in the infarct zone in WT than TNF-/- mice on day 3; (3) matrix metalloproteinase-9 and -2 activity in the infarcted myocardium was significantly higher in WT than in TNF-/ mice on day 3; (4) on day 28 after MI compared with sham, there was a significant decrease in LV developed pressure (74%) and +/-dP/dt(max) (68.3%/65.3%) in WT mice but a less significant decrease in +/-dP/dt(max) (25.8%/28.8%) in TNF-/- mice; (5) cardiac collagen volume fraction was lower in WT than in TNF-/- mice on days 3 and 7 but higher on day 28 compared with TNF-/- mice; and (6) a reduction in myocyte apoptosis in TNF-/- mice occurred on day 28 compared with WT mice. CONCLUSIONS: Elevated local TNF-alpha in the infarcted myocardium contributes to acute myocardial rupture and chronic left ventricle dysfunction by inducing exuberant local inflammatory response, matrix and collagen degradation, increased matrix metalloproteinase activity, and apoptosis. PMID- 15533864 TI - D-4F, an apolipoprotein A-I mimetic peptide, inhibits the inflammatory response induced by influenza A infection of human type II pneumocytes. AB - BACKGROUND: Evidence suggests that apolipoprotein A-I (apoA-I) and HDL play important roles in modulating inflammation. We previously reported that an apoA-I mimetic peptide, D-4F, reduced inflammatory responses to influenza virus in mice. To further define the antiinflammatory activity of D-4F, a human alveolar type II cell line, A549, was used. METHODS AND RESULTS: Cells were either uninfected or infected with influenza A in the presence or absence of D-4F. Cells treated with D-4F were more viable, and virus-induced cytokine production was suppressed by D 4F. Caspases associated with cytokine production were activated after infection but suppressed by D-4F treatment. Infected A549 cells showed dramatic increases in cellular phospholipid secretion into the media. When infected cells were incubated with D-4F, secretion of parent nonoxidized, noninflammatory phospholipids was unaltered, but production of proinflammatory oxidized phospholipids was inhibited. CONCLUSIONS: Type II pneumocytes respond to influenza A infection by activating caspases and secreting cytokines and cellular phospholipids into the extracellular environment, including oxidized phospholipids that evoke inflammatory responses. D-4F treatment inhibited these events. Our results suggest that apoA-I and apoA-I mimetic peptides such as D-4F are antiinflammatory agents that may have therapeutic potential. PMID- 15533865 TI - Effects of the acyl coenzyme A:cholesterol acyltransferase inhibitor avasimibe on human atherosclerotic lesions. AB - BACKGROUND: Inhibition of the acyl coenzyme A:cholesterol acyltransferase (ACAT) enzyme may prevent excess accumulation of cholesteryl esters in macrophages. The ACAT inhibitor avasimibe was shown to reduce experimental atherosclerosis. This study was designed to investigate the effects of avasimibe on human coronary atherosclerosis. METHODS AND RESULTS: This randomized, double-blind, placebo controlled trial assessed the effects of avasimibe at dosages of 50, 250, and 750 mg QD on the progression of coronary atherosclerosis as assessed by intravascular ultrasound (IVUS). All patients received background lipid-lowering therapy if necessary to reach a target baseline LDL level <125 mg/dL (3.2 mmol/L). IVUS and coronary angiography were performed at baseline and repeated after up to 24 months of treatment. Approximately equal percentages of patients across groups received concurrent statin therapy (87% to 89%). The mean total plaque volume at baseline was approximately 200 mm3, and the least squares mean change at end of treatment was 0.7 mm3 for placebo and 7.7, 4.1, and 4.8 mm3 for the avasimibe 50, 250, and 750 mg groups, respectively (adjusted P=0.17 [unadjusted P=0.057], 0.37, and 0.37, respectively). Percent atheroma volume increased by 0.4% with placebo and by 0.7%, 0.8%, and 1.0% in the respective avasimibe groups (P=NS). LDL cholesterol increased during the study by 1.7% with placebo but by 7.8%, 9.1%, and 10.9% in the respective avasimibe groups (P<0.05 in all groups). CONCLUSIONS: Avasimibe did not favorably alter coronary atherosclerosis as assessed by IVUS. This ACAT inhibitor also caused a mild increase in LDL cholesterol. PMID- 15533866 TI - Stromal cell-derived factor-1alpha plays a critical role in stem cell recruitment to the heart after myocardial infarction but is not sufficient to induce homing in the absence of injury. AB - BACKGROUND: After myocardial infarction (MI), bone marrow-derived cells (BMDCs) are found within the myocardium. The mechanisms determining BMDC recruitment to the heart remain unclear. We investigated the role of stromal cell-derived factor 1alpha (SDF-1) in this process. METHODS AND RESULTS: MI produced in mice by coronary ligation induced SDF-1 mRNA and protein expression in the infarct and border zone and decreased serum SDF-1 levels. By quantitative polymerase chain reaction, 48 hours after intravenous infusion of donor-lineage BMDCs, there were 80.5+/-15.6% more BDMCs in infarcted hearts compared with sham-operated controls (P<0.01). Administration of AMD3100, which specifically blocks binding of SDF-1 to its endogenous receptor CXCR4, diminished BMDC recruitment after MI by 64.2+/ 5.5% (P<0.05), strongly suggesting a requirement for SDF-1 in BMDC recruitment to the infarcted heart. Forced expression of SDF-1 in the heart by adenoviral gene delivery 48 hours after MI doubled BMDC recruitment over MI alone (P<0.001) but did not enhance recruitment in the absence of MI, suggesting that SDF-1 can augment, but is not singularly sufficient for, BDMC recruitment to the heart. Gene expression analysis after MI revealed increased levels of several genes in addition to SDF-1, including those for vascular endothelial growth factor, matrix metalloproteinase-9, intercellular adhesion molecule-1, and vascular cell adhesion molecule-1, which might act in concert with SDF-1 to recruit BMDCs to the injured heart. CONCLUSIONS: SDF-1/CXCR4 interactions play a crucial role in the recruitment of BMDCs to the heart after MI and can further increase homing in the presence, but not in the absence, of injury. PMID- 15533867 TI - Presence of left-to-right atrial frequency gradient in paroxysmal but not persistent atrial fibrillation in humans. AB - BACKGROUND: Recent studies have demonstrated spatiotemporal organization in atrial fibrillation (AF), with a left-to-right atrial frequency gradient during AF in isolated sheep hearts. We hypothesized that human AF would also manifest a left-to-right atrial frequency gradient. METHODS AND RESULTS: Thirty-one patients aged 56.7+/-10.5 years with a history of paroxysmal or persistent (>1 month) AF were included. Recordings were made at each pulmonary vein (PV) ostium and simultaneously from the coronary sinus (CS) and posterior right atrium (RA) during AF. Sequential fast Fourier transforms (FFTs) were performed. FFT profiles were analyzed to determine the dominant frequency (DF). There were 18 patients with paroxysmal AF and 13 with persistent AF. In the paroxysmal group, there was a significant left-to-right atrial DF gradient, with DF highest at the PV/left atrial (LA) junction, intermediate at the CS, and lowest in the RA (6.2+/-0.8, 5.5+/-0.7, and 5.1+/-0.6 Hz, respectively; P<0.001). There were no patients in whom DF was greater at the RA than the PV/LA junction. In the persistent group, there was no significant difference between DF recorded from the LA/PV junction, CS, and RA (6.1+/-0.7, 5.8+/-0.6, and 5.8+/-0.6 Hz, respectively; P=NS). CONCLUSIONS: In humans with paroxysmal AF, DFs are highest at the PV/LA junction, intermediate in the CS, and slowest in the posterior RA. These findings agree with animal models that suggest that the posterior LA may play an important role in maintaining paroxysmal AF. The role of the posterior LA in persistent AF requires further study. PMID- 15533868 TI - Right ventricular enlargement on chest computed tomography: a predictor of early death in acute pulmonary embolism. AB - BACKGROUND: In patients with acute pulmonary embolism (PE), rapid and accurate risk assessment is paramount in selecting the appropriate treatment strategy. Right ventricular (RV) enlargement on chest CT has previously been shown to correlate with an unstable hospital course, but its role as a predictor of death is unknown. METHODS AND RESULTS: We evaluated 431 consecutive patients (mean age, 59+/-16 years; 55% women) with acute PE confirmed by multidetector-row chest CT. With the use of multiplanar reformats of axial CT data, CT 4-chamber (4-CH) views were reconstructed and right and left ventricular dimensions (RV(D), LV(D)) were measured. RV enlargement, defined as RV(D)/LV(D) >0.9, was present in 276 (64.0%; 95% CI, 59.5% to 68.6%) patients. Thirty-day mortality rate was 15.6% (95% CI, 11.3% to 19.9%) in patients with and 7.7% (95% CI, 3.5% to 12.0%) without RV enlargement (log rank, P=0.018). The hazard ratio of RV(D)/LV(D) >0.9 for predicting 30-day death was 3.36 (95% CI, 1.13 to 9.97; P=0.029). On multivariable analysis, RV enlargement predicted 30-day death (hazard ratio, 5.17; 95% CI, 1.63 to 16.35; P=0.005) after adjusting for pneumonia (hazard ratio, 2.95; 95% CI, 1.19 to 3.83; P=0.002), cancer (hazard ratio, 2.13; 95% CI, 1.19 to 3.83; P=0.011), chronic lung disease (hazard ratio, 2.00; 95% CI, 1.04 to 3.86; P=0.039), and age (hazard ratio, 1.03; 95% CI, 1.01 to 1.05; P=0.005). CONCLUSIONS: In patients with acute PE, RV enlargement on reconstructed CT 4-CH view helps predict early death. PMID- 15533869 TI - N-terminal pro-B-type natriuretic peptide levels for dynamic risk stratification of patients with acute coronary syndromes. AB - BACKGROUND: Elevated baseline levels of B-type natriuretic peptide (BNP) and the N-terminal fragments of its prohormone, N-terminal-pro-BNP (NT-proBNP), have been associated with adverse long-term outcome in patients with acute coronary syndromes, whereas the prognostic implications of serial NT-proBNP measurements have not been investigated to date. METHODS AND RESULTS: NT-proBNP, troponin T, and C-reactive protein were measured at baseline and at 48 and 72 hours in 1791 patients with non-ST-elevation acute coronary syndromes. Death and myocardial infarction were recorded during 30 days of follow-up. After adjustment for independent predictors of cardiac risk, baseline NT-proBNP levels >250 ng/L were associated with higher event rates (adjusted OR, 3.7; 95% CI, 2.3 to 5.7; P<0.001). In troponin T-negative patients, NT-proBNP identified a subgroup of high-risk patients (OR, 5.9; 95% CI, 2.6 to 13.3; P<0.001). The risk in those patients (7.2%) did not significantly differ from that in troponin T-positive patients (9.8%; P=0.25). Importantly, clinical stabilization without refractory ischemia was associated with a rapid (as soon as 48 hours after onset of symptoms) and significant (48 hours; -24%; 72 hours, -49%; both P<0.001) decline in NT-proBNP levels. In patients with high NT-proBNP baseline levels, lack of a rapid decline in NT-proBNP levels (< or =250 ng/L) was linked to an adverse short term prognosis (OR, 33.7; 95% CI, 8.2 to 138.8; P<0.001). In patients with low NT proBNP baseline levels, a rise in NT-proBNP levels over 72 hours to >250 ng/L was also linked to an adverse 30-day prognosis (OR, 24.0; 95% CI, 8.4 to 68.5; P<0.001). CONCLUSIONS: Neurohumoral activation as evidenced by NT-proBNP appears as a unifying feature that is independent of other biochemical markers (myocardial necrosis, inflammation) and is a powerful and independent determinant of the short-term cardiac risk in patients with acute coronary syndromes. Whether serial measurements of NT-proBNP in patients with ACS may be used to more rapidly identify patients suitable for early discharge or more intensive therapy deserves future prospective studies. PMID- 15533870 TI - Weight-loss-associated induction of peroxisome proliferator-activated receptor alpha and peroxisome proliferator-activated receptor-gamma correlate with reduced atherosclerosis and improved cardiovascular function in obese insulin-resistant mice. AB - BACKGROUND: Weight loss in obese insulin-resistant but not in insulin-sensitive persons reduces coronary heart disease risk. To what extent changes in gene expression are related to atherosclerosis and cardiovascular function is unknown. METHODS AND RESULTS: We studied the effect of diet restriction-induced weight loss on gene expression in the adipose tissue, the heart, and the aortic arch and on atherosclerosis and cardiovascular function in mice with combined leptin and LDL-receptor deficiency. Obesity, hypertriglyceridemia, and insulin resistance are associated with hypertension, impaired left ventricular function, and accelerated atherosclerosis in those mice. Compared with lean mice, peroxisome proliferator-activated receptors (PPAR)-alpha and PPAR-gamma expression was downregulated in obese double-knockout mice. Diet restriction caused a 45% weight loss, an upregulation of PPAR-alpha and PPAR-gamma, and a change in the expression of genes regulating glucose transport and insulin sensitivity, lipid metabolism, oxidative stress, and inflammation, most of which are under the transcriptional control of these PPARs. Changes in gene expression were associated with increased insulin sensitivity, decreased hypertriglyceridemia, reduced mean 24-hour blood pressure and heart rate, restored circadian variations of blood pressure and heart rate, increased ejection fraction, and reduced atherosclerosis. PPAR-alpha and PPAR-gamma expression was inversely related to plaque volume and to oxidized LDL content in the plaques. CONCLUSIONS: Induction of PPAR-alpha and PPAR-gamma in adipose tissue, heart, and aortic arch is a key mechanism for reducing atherosclerosis and improving cardiovascular function resulting from weight loss. Improved lipid metabolism and insulin signaling is associated with decreased tissue deposition of oxidized LDL that increases cardiovascular risk in persons with the metabolic syndrome. PMID- 15533871 TI - Differentiation of intraplaque versus juxtaluminal hemorrhage/thrombus in advanced human carotid atherosclerotic lesions by in vivo magnetic resonance imaging. AB - BACKGROUND: Intraplaque hemorrhage and juxtaluminal hemorrhage/thrombus may differ in cause and clinical implications. This study tested the hypothesis that MRI can distinguish between intraplaque hemorrhage and juxtaluminal hemorrhage/thrombus and investigated the association between hemorrhage and underlying lesion types. METHODS AND RESULTS: Twenty-six patients scheduled for carotid endarterectomy were imaged with a 1.5-T GE scanner by a multicontrast weighted MRI technique. Hemorrhages were identified with previously established MRI criteria, and differentiations were made between intraplaque and juxtaluminal hemorrhage/thrombus. Corresponding histology was used to confirm the magnetic resonance findings. Tissues underlying areas of hemorrhage/thrombus were histologically categorized according to modified American Heart Association criteria. Of 190 matched sections, 140 contained areas of hemorrhage by histology, of which MRI correctly detected 134. The sensitivity and specificity for MRI to correctly identify cross sections that contained hemorrhage were 96% and 82%, respectively. Furthermore, MRI was able to distinguish juxtaluminal hemorrhage/thrombus from intraplaque hemorrhage with an accuracy of 96%. The distribution of lesion types underlying hemorrhages differed significantly (P=0.004). Intraplaque hemorrhage had an underlying lipid-rich type IV/V lesion in 55% of histological sections, whereas juxtaluminal hemorrhage/thrombus had an underlying calcified lesion type VII in 70% of sections. CONCLUSIONS: In vivo high-resolution MRI can detect and differentiate intraplaque hemorrhage from juxtaluminal hemorrhage/thrombus with good accuracy. The association of hemorrhage and lesion types suggests potential differences in origin. Noninvasive MRI therefore provides a possible tool for prospectively studying differences in origin of plaque hemorrhage and the association of plaque progression and instability. PMID- 15533872 TI - Platelet function testing in cardiovascular diseases. PMID- 15533873 TI - Images in cardiovascular medicine. Primary stenting of an anomalous left anterior descending coronary artery originating from the proximal right coronary artery in a patient with acute non-ST-elevation myocardial infarction. PMID- 15533874 TI - Letter regarding article by Yu et al, "Tissue Doppler imaging is superior to strain rate imaging and postsystolic shortening on the prediction of reverse remodeling in both ischemic and nonischemic heart failure after cardiac resynchronization therapy". PMID- 15533875 TI - Congestive heart failure: more bad news from exercising muscle? PMID- 15533876 TI - Cardiovascular effects of the 3 phosphodiesterase-5 inhibitors approved for the treatment of erectile dysfunction. PMID- 15533877 TI - Plasticity in light reactions of photosynthesis for energy production and photoprotection. AB - Plant photosynthesis channels some of the most highly reactive intermediates in biology, in a way that captures a large fraction of their energy to power the plant. A viable photosynthetic apparatus must not only be efficient and robust machinery, but also well integrated into the plant's biochemical and physiological networks. This requires flexibility in its responses to the dramatically changing environmental conditions and biochemical demands. First, the output of the energy-storing light reactions must match the demands of plant metabolism. Second, regulation of the antenna must be flexible to allow responses to diverse challenges that could result in excess light capture and subsequent photoinhibition. Evidence is presented for the interplay of two types of mechanistic flexibility, one that modulates the relative sensitivity of antenna down-regulation to electron flow, and the other, which primarily modulates the output ratio of ATP/NADPH, but also contributes to down-regulation. PMID- 15533878 TI - Co-ordinated gene expression of photosynthetic glyceraldehyde-3-phosphate dehydrogenase, phosphoribulokinase, and CP12 in Arabidopsis thaliana. AB - Photosynthetic glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) interact in the chloroplast stroma through the action of the small peptide CP12. This supramolecular complex concurs with the light dependent modulation in vivo of GAPDH and PRK activities. The expression patterns of several genes potentially involved in the formation of the complex have been studied. The genome of Arabidopsis thaliana includes seven genes for phosphorylating GAPDH isozymes, one PRK gene, and three genes for CP12. The expression of four GAPDH genes was analysed, i.e. GapA-1 and GapB for photosynthetic GAPDH of chloroplasts (NAD(P)-dependent), GapC-1 for cytosolic GAPDH, and GapCp-1 for plastid GAPDH (both NAD-dependent). A similar analysis was performed with PRK and two CP12 genes (CP12-1, CP12-2). The expression of GapA-1, GapB, PRK, and CP12-2 was found to be co-ordinately regulated with the same organ specificity, all four genes being mostly expressed in leaves and flower stalks, less expressed in flowers, and little or not expressed in roots and siliques. The expression of all these genes in leaves was terminated during prolonged darkness or following sucrose treatments, and their transcripts decayed with similar kinetics. At variance with CP12-2, gene CP12-1 appeared to be expressed more in flowers, it was totally insensitive to darkness, and less affected by sucrose. The expression of glycolytic GapC was strong and ubiquitous, insensitive to dark treatments, and unaffected by sucrose. GapCp transcripts were also found to be ubiquitous at lower levels, slowly decreasing in the dark and stable in sucrose treated leaves. The co-ordinated expression of genes GapA-1, GapB, PRK, and CP12 2 is consistent with their specific involvement in the formation of the photosynthetic regulatory complex of chloroplasts. PMID- 15533879 TI - Internal axial light conduction in the stems and roots of herbaceous plants. AB - In order to reveal any roles played by stems and roots of herbaceous plants in responding to the surrounding light environment, the optical properties of the stem and root tissues of 18 herbaceous species were investigated. It was found that light was able to penetrate through to the interior of the stem and was then conducted towards the roots. Light conduction was carried out within the internodes and across the nodes of the stem, and then in the roots from the tap root to lateral roots. Light conduction in both the stem and root occurred in the vascular tissue, usually with fibres and vessels serving as the most efficient axial light conductors. The pith and cortex in many cases were also involved in axial light conduction. Investigation of the spectral properties of the conducted light made it clear that only the spectral region between 710 nm and 940 nm (i.e. far-red and near infra-red light) was the most efficiently conducted in both the stem and the root. It was also found that there were light gradients in the axial direction of the stem or root, and the light intensity generally exhibited a linear attenuation in accord with the distance of conduction. These results revealed that tissues of the stem and root are bathed in an internal light environment enriched in far-red light, which may be involved in phytochrome mediated metabolic activities. Thus, it appears that light signals from above ground directly contribute to the regulation of the growth and development of underground roots via an internal light-conducting system from the stem to the roots. PMID- 15533880 TI - Selective transcriptional down-regulation of anther invertases precedes the failure of pollen development in water-stressed wheat. AB - Water deficit during male meiosis in wheat (Triticum aestivum L.) causes pollen sterility. With a view to identifying the internal trigger for this failure, it was found that water stress specifically impairs the activities of vacuolar and cell-wall invertases in anthers prior to the arrest of pollen development. The enzymes are affected only when water deficit occurs around meiosis. Three invertase cDNAs, two encoding the cell-wall (Ivr1, Ivr3) and one the vacuolar (Ivr5) isoform, were isolated from an anther cDNA library. RNA gel-blot analysis using floral organs of well-watered plants revealed that these genes were expressed preferentially, though not exclusively, in anthers. Semi-quantitative RT-PCR demonstrated that transitory water deficit during meiosis selectively down regulated the transcription of two of the three genes, one encoding the vacuolar (Ivr5) and the other a cell-wall (Ivr1) isoform, without affecting the Ivr3 message. Their expression did not recover upon resumption of watering. Another homologue of Ivr1 was also down-regulated, but only during the post-stress period. The stress effects on invertase transcripts were consistent with those on the developmental profiles of the corresponding enzyme activities. In situ hybridization revealed that the stress-sensitive invertase genes, unlike an insensitive one, were expressed within the microspores. No evidence for an invertase inhibitor under stress was found. Together the results show that the decline in invertase activity is probably regulated primarily at the transcriptional level in a gene- and cell-specific manner. PMID- 15533881 TI - Genotypic variation of the response to cadmium toxicity in Pisum sativum L. AB - This work evaluates the (cor-)relations between selected biochemical responses to toxic Cd and the degree of Cd sensitivity in a set of pea genotypes. Ten genotypes were analysed that differ in their growth response to Cd when expressed as root or shoot tolerance indices (TIs). Concentrations of non-protein thiols (NPTs) and malondialdehyde (MDA), activity of chitinase, peroxidase (POX), and catalase significantly increased in all pea genotypes treated with Cd. Cd sensitivity of genotypes was correlated with relative increases in MDA concentration as well as activities of chitinase and POX, suggesting similar Cd stress effects. Activities of ascorbate peroxidase (APX) decreased, but concentrations of glutathione (GSH) increased in the less Cd-sensitive genotypes. Differences in root and leaf contents of Cd revealed no correlation with TI, metabolic parameters, and enzyme activities in Cd-treated plants, respectively, except that shoot Cd concentration positively correlated with shoot chitinase activity. Toxic Cd levels inhibited uptake of nutrient elements such as P, K, S, Ca, Zn, Mn, and B by plants in an organ- and genotype-specific manner. Cd sensitivity was significantly correlated with decreased root Zn concentrations. The results show both similarities, as well as distinct features, in Cd toxicity expression in genotypes of one species, suggesting that independent and multi factorial reactions modulate Cd sensitivity on the low-tolerance level of plants. The study illustrates the biochemical basis of earlier detected genotypic variation in Cd response. PMID- 15533882 TI - Metabolic engineering of high carotenoid potato tubers containing enhanced levels of beta-carotene and lutein. AB - In order to enhance the carotenoid content of potato tubers, transgenic potato plants have been produced expressing an Erwinia uredovora crtB gene encoding phytoene synthase, specifically in the tuber of Solanum tuberosum L. cultivar Desiree which normally produces tubers containing c. 5.6 microg carotenoid g(-1) DW and also in Solanum phureja L. cv. Mayan Gold which has a tuber carotenoid content of typically 20 microg carotenoid g(-1) DW. In developing tubers of transgenic crtB Desiree lines, carotenoid levels reached 35 microg carotenoid g( 1) DW and the balance of carotenoids changed radically compared with controls: beta-carotene levels in the transgenic tubers reached c. 11 microg g(-1) DW, whereas control tubers contained negligible amounts and lutein accumulated to a level 19-fold higher than empty-vector transformed controls. The crtB gene was also transformed into S. phureja (cv. Mayan Gold), again resulting in an increase in total carotenoid content to 78 microg carotenoid g(-1) DW in the most affected transgenic line. In these tubers, the major carotenoids were violaxanthin, lutein, antheraxanthin, and beta-carotene. No increases in expression levels of the major carotenoid biosynthetic genes could be detected in the transgenic tubers, despite the large increase in carotenoid accumulation. Microarray analysis was used to identify a number of genes that were consistently up- or down-regulated in transgenic crtB tubers compared with empty vector controls. The implications of these data from a nutritional standpoint and for further modifications of tuber carotenoid content are discussed. PMID- 15533883 TI - Analysis of two alleles of the urease gene from potato: polymorphisms, expression, and extensive alternative splicing of the corresponding mRNA. AB - Globally, urea is the most widely used nitrogen fertilizer and is made accessible to plants via the urease reaction. However, sequence information for the plant enzyme is scarce. A cDNA encoding urease from soybean (Glycine max) has been cloned and sequence information has been obtained for two alleles (11 and 19 kbp, respectively) of the potato (Solanum tuberosum, cv. Desiree) urease gene and the corresponding cDNAs. It was found that urease is encoded by a single copy gene in several solanaceous species, and maps to chromosome V in potato. By contrast, the presence of two urease genes was reported for soybean. Comparative analysis of 11 kbp overlapping allelic DNA allowed the quantification of single nucleotide polymorphisms and revealed the presence of a truncated Ty1-copia retrotransposon in one of the alleles. Both alleles contained a copy of a terminal-repeat retrotransposon in miniature (TRIM). 25-50% of urease pre-mRNAs from both alleles were alternatively spliced in a variety of different ways. The retrotransposons had no effect on splicing. While urease is expressed in all tissues tested, its mRNA and protein are of low abundance. The TATA-less urease promoter appears to drive low-level housekeeping transcription. An in silico analysis showed that eukaryotic urease protein sequences are very similar to sequences from prokaryotic enzymes, conserving all residues of known functional importance. It is therefore likely that all known ureases are structurally similar, employing the same catalytic mechanism. PMID- 15533884 TI - Annual and seasonal variation of sap flow and conductance of pine trees grown in elevated carbon dioxide and temperature. AB - Measurements of sap flow, crown structure, and microclimate were used to estimate the transpiration of individual 30-year-old Pinus sylvestris L. trees grown in elevated temperature and CO2. The trees were enclosed in closed-top chambers and exposed either to current ambient conditions (CON), or elevated CO2 (+350 micromol mol(-1); EC), or elevated temperature (+2 to +6 degrees C; ET) or a combination of EC and ET (ECT) since 1996, and the measurements were made from 1999 to 2001. EC significantly increased annual sap flow per tree (Ft.m) by 14% in 1999, but reduced it by 13% in 2000 and 16% in 2001. The CO2-induced increase in Ft.m in 1999 was due to a large increase in foliage area of trees, which more than compensated for a small decrease in crown conductance (Gc). The CO2-induced decreases in Ft.m in 2000 and 2001 resulted from a pronounced decline in Gc, which was much greater than the increase in foliage area. The CO2-induced increase in sensitivity of Gc at high vapour pressure deficit (VPD) did not alter the general response of sap flow to CO2 enrichment, but it did affect the diurnal courses of sap flow on some days during the main growing season (days 150-240). ET increased Ft.m by 53%, 45%, and 57% in 1999, 2000, and 2001, respectively, attributable to the combined effects of greater foliage area and maximum crown conductance, lower stomatal sensitivity to high VPD, and higher transpiration demand relative to the control treatments. There was no significant interaction between CO2 and temperature on sap flow, because ECT entailed approximately similar patterns of sap flow to ET, suggesting that the temperature played a dominate role in the case of ECT under boreal climate conditions. PMID- 15533885 TI - Functional analysis of homologous and heterologous promoters in strawberry fruits using transient expression. AB - The isolation and characterization of fruit-specific promoters are critical for the manipulation of the nutritional value and quality of fruits by genetic engineering. The analysis of regulatory sequences of many ripening-related genes has remained elusive for many species due to their low transformation efficiency and/or lengthy regeneration of a small number of transgenic plants. Strawberry is an important crop and represents one of the most widely studied non-climacteric model systems. However, until recently, its difficult regeneration has limited the functional study of promoters by stable transformation. A protocol based on biolistic transient transformation has been developed in order to study the function of promoters in a fast and efficient manner in strawberry fruits. The protocol has been applied to the study of the GalUR promoter, a gene involved in the biosynthesis of vitamin C in this fruit. The activity of the GalUR promoter is restricted to the fruit, being strictly dependent on light. The analysis of deletion series revealed the presence of a minimum activation region 397 bp upstream of the gene with a putative G-box motif, and a negative regulatory region between -397 and -518 bp, where an I-box was identified. The transient assay has been used to study the activity of the tomato polygalacturonase and the pepper fibrillin promoters in strawberry fruits. Whereas slight activity was observed with the fibrillin promoter, no significant activity was found with the polygalacturonase promoter. The GalUR promoter in transiently transformed ripe tomato fruits showed no activity, indicating the presence of regulatory sequences specific for its function in strawberry fruit. PMID- 15533888 TI - Inhibition of ATP-sensitive potassium channels by haloperidol. AB - Chronic haloperidol treatment has been associated with an increased incidence of glucose intolerance and type-II diabetes mellitus. We studied the effects of haloperidol on native ATP-sensitive potassium (K(ATP)) channels in mouse pancreatic beta cells and on cloned Kir6.2/SUR1 channels expressed in HEK293 cells. The inhibitory effect of haloperidol on the K(ATP) channel was not mediated via the D2 receptor signaling pathway, as both D2 agonists and antagonists blocked the channel. K(ATP) currents were studied using the patch clamp technique in whole-cell and outside-out patch configurations. Addition of haloperidol to the extracellular solution inhibited the K(ATP) conductance immediately, in a reversible and voltage-independent manner. Haloperidol did not block the channel when applied intracellularly in whole-cell recordings. Haloperidol blocked cloned Kir6.2/SUR1 and Kir6.2DeltaC36 K(ATP) channels expressed in HEK cells. This suggests that the drug interacts with the Kir6.2 subunit of the channel. The IC(50) for inhibition of the K(ATP) current by haloperidol was 1.6 microM in 2 mM extracellular K(+) concentration ([K(+)](o)) and increased to 23.9 microM in 150 mM [K(+)](o). The Hill coefficient was close to unity, suggesting that the binding of a single molecule of haloperidol is sufficient to close the channel. Haloperidol block of K(ATP) channels may contribute to the side effects of this drug when used therapeutically. PMID- 15533889 TI - Platelet-leukocyte aggregation induced by PAR agonists: regulation by nitric oxide and matrix metalloproteinases. AB - Platelet-leukocyte aggregation (PLA) links haemostasis to inflammation. The role of nitric oxide (NO) and matrix metalloproteinases (MMP-1, -2, -3, -9) in PLA regulation was studied. Homologous human platelet-leukocyte suspensions were stimulated with thrombin (0.1-3 nM) and other proteinase activated receptor activating peptides (PAR-AP), including PAR1AP (0.5-10 microM), PAR4AP (10-70 microM), and thrombin receptor-activating peptide (1-35 microM). PLA was studied using light aggregometry with simultaneous measurement of oxygen-derived free radicals, dual colour flow cytometry, and phase-contrast microscopy. The release of NO was measured using a porphyrinic nanosensor, while MMPs were investigated by Western blot, substrate degradation assays, immunofluorescence microscopy, and flow cytometry. The levels of P-selectin and microparticles (MP) in PLA were measured by flow cytometry. PLA was also characterized using pharmacological agents: S-nitroso-glutathione (GSNO, 0.01-10 microM), 1H-Oxadiazole quinoxalin-1 one (ODQ, 1 microM), N(G)-L-nitro-L-arginine methyl ester (L-NAME, 100 microM) and compounds that modulate the actions of MMPs such as phenanthroline (100 microM), monoclonal anti-MMP antibodies, and purified MMPs. PAR agonists concentration-dependently induced PLA, an effect associated with the release of microparticles (MP) and the translocation of P-selectin to the platelet surface. NO and radicals were also released during PLA. Inhibition of NO bioactivity by the concomitant release of free radicals or by the treatment with L-NAME or ODQ stimulated PLA, while pharmacological administration of GSNO decreased PLA. PAR agonist-induced PLA resulted in the liberation of MMP-1, -2, -3, and -9. During PLA, MMPs were present on the cell surface, as shown by flow cytometry and immunofluorescence. PLA led to the activation of latent MMPs to active MMPs, as shown by Western blot and substrate degradation assays. Inhibition of MMPs actions by phenanthroline and by the antibodies attenuated PLA. In contrast, purified active, but not latent, MMPs amplified thrombin-induced PLA. It is concluded that NO and MMP-1, -2, -3, and -9 play an important role in regulation of PAR agonist-induced PLA. PMID- 15533890 TI - GW9662, a potent antagonist of PPARgamma, inhibits growth of breast tumour cells and promotes the anticancer effects of the PPARgamma agonist rosiglitazone, independently of PPARgamma activation. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma), a member of the nuclear receptor superfamily, is activated by several compounds, including the thiazolidinediones. In addition to being a therapeutic target for obesity, hypolipidaemia and diabetes, perturbation of PPARgamma signalling is now believed to be a strategy for treatment of several cancers, including breast. Although differential expression of PPARgamma is observed in tumours compared to normal tissues and PPARgamma agonists have been shown to inhibit tumour cell growth and survival, the interdependence of these observations is unclear. This study demonstrated that the potent, irreversible and selective PPARgamma antagonist GW9662 prevented activation of PPARgamma and inhibited growth of human mammary tumour cell lines. Controversially, GW9662 prevented rosiglitazone-mediated PPARgamma activation, but enhanced rather than reversed rosiglitazone-induced growth inhibition. As such, these data support the existence of PPARgamma independent pathways and question the central belief that PPARgamma ligands mediate their anticancer effects via activation of PPARgamma. PMID- 15533891 TI - Inhibition of calcium-calmodulin kinase restores nitric oxide production and signaling in submandibular glands of a mouse model of salivary dysfunction. AB - Nitric oxide is an intracellular and diffusible messenger of neurotransmitters involved in salivary secretion, as well as an inflammatory mediator in salivary gland diseases. It is synthesized by three different isoforms of nitric oxide synthase (NOS), each subject to a fine transcriptional, post-transcriptional and/or post-translational regulation. Our purpose was to study the possible mechanisms leading to NOS downregulation in submandibular glands of normal mice and in the nonobese diabetic (NOD) mouse model of salivary dysfunction with lower NOS activity. NOS activity and cGMP accumulation were determined by radioassays in submandibular glands of both mice in the presence of the protein kinase inhibitors KN-93 and bisindolylmaleimide. NOS I mRNA and protein expression and localization were assessed by RT-PCR, Western blot and immunohistochemistry. A downregulatory effect of calcium-calmodulin kinase II (CaMK II) on NOS activity in submandibular glands of both NOD and BALB/c mice was observed. Our results are consistent with a physiological regulation of NOS activity by this kinase but not by PKC in normal BALB/c mice. They are also supportive of a role for CaMK II in the lack of detectable NOS activity in submandibular glands of NOD mice. KN-93 also restored cGMP accumulation in NOD submandibular glands. The downregulation of NOS in NOD mice seems to be mainly mediated by this kinase rather than the result of a lower expression or different cellular localization of the enzyme. It was not related to different substrate or cofactors availability either. PMID- 15533893 TI - Genetics in clinical cancer care: a promise unfulfilled among minority populations. PMID- 15533892 TI - The intestinal anti-inflammatory effect of quercitrin is associated with an inhibition in iNOS expression. AB - Quercitrin, 3-rhamnosylquercetin, is a bioflavonoid with antioxidant properties, which exerts anti-inflammatory activity in experimental colitis. In the present study, different in vivo experiments were performed in order to evaluate the mechanisms of action involved in this effect, with special attention to its effects on proinflammatory mediators, including nitric oxide (NO). Experimental colitis was induced in female Wistar rats by incorporation of dextran sodium sulfate (DSS) in drinking water. Oral treatment of quercitrin (1 or 5 mg kg(-1) day(-1)) to colitic rats ameliorated the evolution of the inflammatory process induced when administered in a preventative dosing protocol. When quercitrin (1 mg kg(-1) day(-1)) was administered on established colitis, it facilitated the recovery of the inflamed mucosa. The beneficial effects exerted by quercitrin were evidenced both histologically and biochemically, and were associated with an improvement in the colonic oxidative status, altered as a consequence of the colonic insult induced by DSS. In addition, a reduction of colonic NO synthase activity was observed, probably related to a decreased expression in the inducible form of the enzyme via downregulation in the colonic activity of the nuclear factor-kappaB. Immunohistochemical studies showed that quercitrin treatment reduced macrophage and granulocyte infiltration in the inflamed tissue. PMID- 15533894 TI - Circulating endogenous retinoic acid concentrations among participants enrolled in a randomized placebo-controlled clinical trial of retinyl palmitate. AB - Retinoids have been studied extensively for their chemopreventive properties. The biological activity of retinoids is acquired through their conversion to retinoic acid (RA). Characterization of endogenous circulating RA concentrations after supplementation with vitamin A over longer time periods has not been done previously. Our investigation was conducted to determine whether vitamin A (retinyl palmitate) supplementation significantly increases circulating RA concentrations of all-trans-, 9-cis-, and 13-cis-RA. Using plasma samples from 41 participants enrolled in a randomized clinical trial of placebo, 25,000, 50,000, or 75,000 IU supplemental retinyl palmitate daily, high-performance liquid chromatography analyses were conducted for concentrations of three RA isomers. Seven plasma samples were analyzed for each participant over a 16-month period. Based on an intention-to-treat analysis, results obtained using linear mixed models showed that supplementation with retinyl palmitate statistically significantly increased concentrations of all three RA isomers from baseline levels. This study suggests that supplementation with retinyl palmitate is an effective means to increase circulating all-trans, 9-cis-, and 13-cis-RA concentrations among humans. PMID- 15533895 TI - Temporal variation and identification of factors associated with endogenous retinoic acid isomers in serum from Brazilian women. AB - OBJECTIVE: Retinoids (natural and synthetic derivatives of vitamin A) have cancer chemotherapeutic and chemopreventive activities. Retinoic acid (RA) treatment has been associated with significant regression of preneoplastic lesions. However, serious toxicity associated with some therapies has made long-term chemoprevention in healthy populations unfeasible. Recently, serum RA has been shown to increase in response to oral retinol (vitamin A) supplementation. Here, we assess the variability of circulating RA levels and the lifestyle, demographic, and nutritional factors that explain such variability. METHOD: Total RA concentration and the concentrations of RA isomers (all-trans-RA, 13-cis-RA, and 9-cis-RA) were measured by high-pressure liquid chromatography in serum samples obtained 4 months apart from 502 women participating in the Ludwig-McGill Cohort (Sao Paulo, Brazil). RESULTS: The relative abundance of the three RA isomers was similar for each visit (baseline and month 4), with 13-cis-RA having the highest concentrations followed by 9-cis-RA and all-trans-RA. The within person variability of total RA and individual isomers was low. Using multivariate logistic regression models (upper tertile versus low/middle tertile of serum RA), we found that age, race, oral contraceptive use, total number of pregnancies, and season of initial blood draw were significantly associated with at least one endogenous RA isomer level. All endogenous RA isomers were positively associated with serum retinol, beta-carotene, and beta-cryptoxanthin levels. CONCLUSION: These results have implications for the design of future epidemiologic studies focused on assessing RA-disease association and intervention studies aimed at modulating RA levels. PMID- 15533896 TI - Peroxisome proliferator-activated receptor-gamma suppresses cyclooxygenase-2 expression in human prostate cells. AB - Recent studies have found that cyclooxygenase-2 (COX-2) protein expression was low and inducible with cytokines in prostate cancer cells (in the absence of serum) and that, in contrast, COX-2 expression was high in normal prostate epithelial cells (EC). Peroxisome proliferator-activated receptor-gamma (PPAR gamma) was expressed at high levels in the prostate cancer cell line PC-3 but not in ECs. In contrast to previous findings by others, PPAR-gamma ligands did not induce PPAR-gamma expression in EC or PC-3. The present study examined the relationship between PPAR-gamma and COX-2 expression patterns in EC and PC-3 in the presence and absence of serum and/or the PPAR-gamma agonist 15-deoxy Delta12,14-prostaglandin J(2) (15d-PGJ(2)). We also evaluated the effects that the forced expression of PPAR-gamma1 and PPAR-gamma2 had on COX-2 in ECs. We found that expression of PPAR-gamma and COX-2 protein was inversely correlated in ECs and PC-3. Low COX-2 expression in PC-3 was up-regulated by serum, and 15d PGJ(2) blocked serum-induced COX-2 expression and activity in a dose-dependent manner. 15d-PGJ(2) had no effect on COX-2 expression in ECs or PPAR-gamma expression in either cell type. However, forced expression of PPAR-gamma1 or PPAR gamma2 in ECs suppressed the high level of endogenous COX-2. This effect was not isoform specific and was augmented by 15d-PGJ(2). The present study showed that PPAR-gamma activation can be an important regulator of COX-2 in prostate cells and may be an important target for prostate cancer chemoprevention. PMID- 15533897 TI - Differential peroxisome proliferator-activated receptor-gamma isoform expression and agonist effects in normal and malignant prostate cells. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is being studied intensively for its role in carcinogenesis and in mediating the effects of prostate cancer treatment and prevention drugs. Prostate cancers express abundant and higher constitutive levels of PPAR-gamma than do normal prostate cells and are growth inhibited by ligand activation of PPAR-gamma. However, little is known about the role of PPARs in tumorigenesis or in normal prostate epithelial cells (EC). We examined the expression, phosphorylation patterns, and functions of the human PPAR (hPPAR)-gamma1 and hPPAR-gamma2 isoforms in normal prostate ECs to determine if activation of the receptor is sufficient for PPAR-gamma ligand activity in prostate cells. We found that ECs did not express either PPAR-gamma1 or PPAR-gamma2 protein and were not sensitive to growth inhibition by the PPAR gamma ligand 15-deoxy-Delta12,14-prostaglandin J(2) (15d-PGJ(2)). In contrast, prostate cancer cells (PC-3), which express PPAR-gamma1 receptor isoform, are growth inhibited by PPAR-gamma ligand. Forced expression of hPPAR-gamma1 or hPPAR gamma2 made ECs sensitive to 15d-PGJ(2) and led to reduced cellular viability. The direct repeat-1 promoter containing PPAR response elements was transactivated in ECs expressing exogenous PPAR-gamma1 or PPAR-gamma2, indicating that either isoform can be active in these cells. 15-Lipoxygenase-2, expressed at high levels in ECs, was down-regulated by transfecting PPAR-gamma expression construct (either gamma1 or gamma2 isoform) into ECs. Addition of PPAR-gamma ligand 15 hydroxyeicosatetraenoic acid in the presence of PPAR-gamma expression caused further down-regulation of 15-lipoxygenase-2. Our data illustrate that a PPAR gamma ligand (15d-PGJ(2)) activates PPAR-gamma1 and selectively induces cell death in human prostate cancer cells but not in normal prostate ECs. These findings have important implications for the development of PPAR-gamma-targeting agents that prevent or treat prostate cancer and spare normal prostate cells. PMID- 15533898 TI - CD151 protein expression predicts the clinical outcome of low-grade primary prostate cancer better than histologic grading: a new prognostic indicator? AB - OBJECTIVE: CD151 is the first member of the tetraspanin family to be associated as a promoter of human tumor metastasis. However, its biological function and expression phenotype among different tumors has not been well investigated. METHOD: Tissue specimens from 76 primary prostate cancers and 30 benign prostate hyperplasia (BPH) controls were obtained from the Department of Anatomical Pathology at the Austin and Repatriation Medical Centre (now Austin Health) from 1984 to 1993. We used quantitative immunohistochemical analysis to measure CD151 protein expression. Analyses of differences among BPH and prostate cancer groups were done with one-way ANOVA and Newman-Keuls test. The Kaplan-Meier method and the log-rank test were used to estimate the overall survival. RESULTS: CD151 expression was found to be significantly higher in prostate cancer specimens compared with BPH specimens (P < 0.001). Poorly differentiated cancers expressed the strongest staining, whereas well-differentiated cancers expressed the weakest staining for CD151 (P < 0.001). The overall survival rate for cases in which CD151 expression was reduced was significantly higher than for cases in which CD151 expression was increased (P = 0.039) especially in well and moderately differentiated cancers (P = 0.014). This effect was independent of the patients' age or preoperative prostate-specific antigen values and superior in the predictive ability of the Gleason score. CONCLUSIONS: CD151 has an increasing expression pattern in prostate cancer progression, and higher levels of CD151 are associated with poorer prognosis. CD151 had better predicting value for the clinical outcome of prostate cancer patients than does the traditional histologic grading method (Gleason grading). PMID- 15533899 TI - Differential effects of Helicobacter pylori eradication on oxidative DNA damage at the gastroesophageal junction and at the gastric antrum. AB - BACKGROUND AND AIM: Helicobacter pylori-associated gastritis causes accumulation of reactive oxygen species in the mucosal compartment. This prospective study evaluates DNA oxidative damage in biopsy samples obtained from both the antrum and the gastroesophageal junction (GEJ) before and after H. pylori eradication. PATIENTS AND METHODS: Thirty-two consecutive H. pylori-positive patients underwent endoscopy with multiple biopsy sampling (i.e., antrum, incisura angularis, fundus, and cardia at the GEJ). After H. pylori eradication, 32 patients underwent a checkup endoscopy (mean interval, 5.7 months); in a subgroup of 13 subjects, a third endoscopy procedure was also performed (mean interval, 18 months). Additional biopsy samples (two from the antrum and two from the GEJ) were used to assess 8-hydroxydeoxyguanosine (8OHdG) levels using both high pressure liquid chromatography with electrochemical detector and ELISA. RESULTS: In the antral compartment, no significant modifications of 8OHdG levels were assessed after H. pylori eradication. Conversely, following eradication, 8OHdG levels significantly increased (high-pressure liquid chromatography with electrochemical detector, P = 0.04; ELISA method, P = 0.05) in biopsy samples taken from the GEJ, and a further increase was documented in the subgroup of patients who underwent a third endoscopy (P = 0.01). The increasing trend was more relevant in patients in whom H. pylori-cagA-positive strains were eradicated and in those affected by hiatal hernia. CONCLUSIONS: The levels of DNA adducts in the antral mucosa are not modified by H. pylori eradication; conversely, H. pylori eradication significantly increases the oxidative adducts at the GEJ. The clinical and biological importance of this situation and whether and how it relates to a higher risk of precancerous lesions is open to debate. PMID- 15533900 TI - Benzene exposure assessed by metabolite excretion in Estonian oil shale mineworkers: influence of glutathione s-transferase polymorphisms. AB - Measurement of urinary excretion of the benzene metabolites S-phenylmercapturic acid (S-PMA) and trans,trans-muconic acid (t,t-MA) has been proposed for assessing benzene exposure, in workplaces with relatively high benzene concentrations. Excretion of S-PMA and t,t-MA in underground workers at an oil shale mine were compared with the excretion in workers engaged in various production assignments above ground. In addition, possible modifying effects of genetic polymorphisms in glutathione S-transferases T1 (GSTT1), M1 (GSTM1), and P1 (GSTP1) on the excretion of S-PMA and t,t-MA were investigated. Fifty underground workers and 50 surface workers participated. Blood samples and three urine samples were collected from each worker: (a) a preshift sample collected the morning after a weekend, (b) a postshift sample 1 collected after the first shift, and (c) a postshift sample 2 collected after the last shift of the week. Personal benzene exposure was 114 +/- 35 mug/m(3) in surface workers (n = 15) and 190 +/- 50 mug/m(3) in underground workers (n = 15) in measurements made prior to the study. We found t,t-MA excretion to be significantly higher in underground workers after the end of shifts 1 and 2 compared with the corresponding surface workers. The same picture, although not significant, was seen for S-PMA excretion. Excretion of S-PMA and t,t-MA was found to increase significantly during the working week in underground workers but not in those employed on the surface. Both t,t-MA and S-PMA excretion were significantly higher in smokers compared with nonsmokers. Subjects carrying the GSTT1 wild-type excreted higher concentrations of S-PMA than subjects carrying the null genotype, suggesting that it is a key enzyme in the glutathione conjugation that leads to S-PMA. The results support the use of benzene metabolites as biomarkers for assessment of exposure at modest levels and warrant for further investigations of health risks of occupational benzene exposure in shale oil mines. PMID- 15533901 TI - Effects of a 2-year randomized soy intervention on sex hormone levels in premenopausal women. AB - OBJECTIVE: Several epidemiologic studies have described protective effects of soy consumption against breast cancer. The goal of this trial among premenopausal women was to examine the effect of soy foods on menstrual cycle length and circulating sex hormone levels. METHODS: This 2-year dietary intervention randomized 220 healthy premenopausal women. The intervention group consumed two daily servings of soy foods containing approximately 50 mg of isoflavones; the control group maintained their regular diet. Five blood samples (obtained in months 0, 3, 6, 12, and 24) were taken 5 days after ovulation as determined by an ovulation kit. The serum samples were analyzed for estrone, estradiol, sex hormone binding globulin, androstenedione, and progesterone by immunoassay. RESULTS: At baseline, both groups had similar demographic, anthropometric, and nutritional characteristics. The dropout rates of 15.6% (17 of 109) in the intervention group and 12.6% (14 of 111) in the control group did not differ significantly. According to soy intake logs, 24-hour recalls, and urinary isoflavone excretion, the women closely adhered to the study regimen. Menstrual cycles became slightly shorter in both groups but did not differ by group. Mixed general linear models indicated no significant intervention effect on any of the serum hormones. However, androstenedione and progesterone decreased significantly over time in both groups. CONCLUSIONS: The results of this study suggest that the preventive effects of soy on breast cancer risk in premenopausal women may not be mediated by circulating sex hormone levels. Different mechanisms of actions or effects of exposure earlier in life are alternate hypotheses that require further investigation. PMID- 15533902 TI - Lack of effect of celecoxib on prostaglandin E2 concentrations in nipple aspirate fluid from women at increased risk of breast cancer. AB - BACKGROUND: Cyclooxygenase enzymes (COX-1, COX-2, and COX-3) convert arachidonic acid to prostaglandins, prostacyclins, thromboxanes, and other hydroxy fatty acids. Among these, prostaglandin E(2) (PGE(2)) has tumor growth-promoting activity. The COX-2 isoform is the primary enzyme involved in PGE(2) production in cancerous tissue. OBJECTIVE/HYPOTHESIS: We administered the COX-2 inhibitor celecoxib (200 mg b.i.d.) to women at increased breast cancer risk. Our hypothesis was that PGE(2) would be secreted in breast nipple aspirate fluid (NAF), that levels in NAF would be higher than in corresponding plasma, and that celecoxib would decrease PGE(2) levels in NAF (reflecting a decreased breast tissue eicosanoid production) and plasma. SPECIFIC AIM: To determine if PGE(2) concentrations in NAF and plasma decrease after a 2-week course of celecoxib and then return to baseline 2 weeks after stopping the medication (washout). STUDY DESIGN: NAF and plasma were collected before celecoxib treatment, 2 weeks after taking celecoxib, and 2 weeks after washout. Each woman served as her own control. RESULTS: PGE(2) concentrations in NAF and plasma were detectable in samples using two measurement techniques. On average, NAF PGE(2) levels were 81 fold higher in NAF than in matched plasma. Technically, there were differences in PGE(2) concentrations measured in similar fluids depending on the assay technique used (RIA versus chemiluminescence immunoassay). There were no significant decreases in PGE(2) concentrations after celecoxib administration. CONCLUSIONS: PGE(2) can be measured in NAF. PGE(2) levels are concentrated in NAF when compared with matched plasma samples. Celecoxib 200 mg b.i.d. does not appear to significantly decrease PGE(2) concentrations in NAF and plasma. PMID- 15533903 TI - Risk factors for breast cancer associated with mammographic features in Singaporean chinese women. AB - BACKGROUND: Mammographic density has been found to be a strong risk factor for breast cancer and to be associated with age, body weight, parity, and menopausal status. Most studies to date have been carried out in Western populations. The purpose of the study described here was to determine in a cross-sectional study in a Singaporean Chinese population the demographic, menstrual, reproductive, and anthropometric factors that are associated with quantitative variations in age adjusted percentage mammographic densities and to examine the association of these factors with the dense and nondense areas of the mammogram. METHOD: We used mammograms and questionnaire data collected from subjects in the Singapore Breast Screening Project. Women ages 45 to 69 years participated and 84% of those screened were Chinese. Mammograms were digitized and percentage density was measured and analyzed in relation to the questionnaire data. RESULTS: Percentage mammographic density was associated with several risk factors for breast cancer, most of them also associated, in opposite directions, with the dense and nondense components of the image. Percentage density was associated with age and weight (both negatively), height and age at first birth (both positively), and number of births and postmenopausal status (both negatively). Percentage density was weakly associated with a previous breast biopsy but was not associated with age at menarche or menopause, with use of hormones, or with a family history of breast cancer. CONCLUSION: Percentage mammographic density in Singaporean Chinese women has similar associations with risk factors for breast cancer to those seen in Caucasians. PMID- 15533904 TI - Serum insulin-like growth factors, insulin-like growth factor binding proteins, and breast cancer risk in postmenopausal women. AB - BACKGROUND: Studies have shown a positive association between serum insulin-like growth factor (IGF)-I and breast cancer risk in premenopausal but not postmenopausal women. IGF-II and estrogen receptor (ER) status has never been investigated. We examined the association between IGF-I, IGF-II, IGF binding protein (IGFBP)-2, IGFBP-3, and IGFBP-3 protease activity and breast cancer risk in postmenopausal women, taking ER status of the breast cancer into consideration. METHODS: We conducted this case-cohort study within a Danish follow-up study based on 24,697 postmenopausal women. We identified 411 cases with breast cancer and a matched control group including 397 cohort members. We estimated breast cancer risk using Cox regression analysis with adjustment for known breast cancer risk factors. RESULTS: We observed no association for IGF-I but a positive association between levels of IGFBP-3 and breast cancer risk. Per 500 units higher levels of IGFBP-3, an incidence rate ratio [IRR; 95% confidence interval (95% CI)] of 1.14 (1.00-1.30) was estimated. For ER-positive breast cancer, the IRR (95% CI) was 1.18 (1.05-1.33). IGFBP-3 protease activity was not associated with breast cancer risk. Per 275 units higher levels of IGF-II, an IRR (95% CI) of 1.35 (1.10-1.66) was observed for ER-positive tumors, whereas IGFBP-2 was not associated with breast cancer risk. Adjustment for potential confounders did not change the risk estimate. There was no association between IGF-I, IGF-II, IGFBP-2, or IGFBP-3 and risk of ER-negative breast cancer. CONCLUSION: Serum IGFBP-3 and IGF-II levels were positively associated with ER-positive breast cancer risk. This may suggest an important relationship among IGFs, IGFBPs, the ER system, and breast cancer development in postmenopausal women. PMID- 15533905 TI - How strong is the association between CAG and GGN repeat length polymorphisms in the androgen receptor gene and prostate cancer risk? AB - OBJECTIVE: Although narrative reviews have suggested an association between (CAG)n and (GGN)n polymorphisms in the AR gene and prostate cancer, it has never been quantified systematically. The purpose of this meta-analysis was to provide relative and absolute quantitative summary estimates with sufficient power. METHOD: Publications were identified through database searches for epidemiologic studies published until February 2004. For each study, mean differences in repeat length between cases and controls were calculated as well as continuous odds ratios (OR) per one CAG or GGN repeat decrement and discrete ORs to compare prostate cancer risk in men with short CAG repeats (21 repeats) and short GGN repeats (16 repeats). The study-specific estimates were combined by random effects metaregression analyses. RESULTS: Nineteen case-control studies were included in this review comprising a total of 4,274 cases and 5,275 controls. Prostate cancer cases had on average 0.26 fewer CAG repeats and 0.09 fewer GGN repeats than controls. The continuous ORs of prostate cancer per one repeat decrement were 1.02 and 1.01 for CAG and GGN repeats, respectively. The summary discrete OR (95% confidence interval) were 1.19 (1.07-1.31) and 1.31 (1.06-1.61) for CAG and GGN repeat polymorphisms, respectively. CONCLUSION: Although the presence of shorter repeats seemed to be modestly associated with prostate cancer risk, the absolute difference in number of repeats between cases and controls is <1 repeat. We question whether such a small difference is enough to yield measurable biological impact in prostate carcinogenesis. PMID- 15533906 TI - Prediagnostic levels of serum micronutrients in relation to risk of gastric cancer in Shanghai, China. AB - Data on blood levels of specific carotenoids and vitamins in relation to gastric cancer are scarce. Little is known about the relationship between prediagnostic serum levels of carotenoids other than beta-carotene and risk of gastric cancer especially in non-Western populations. Prediagnostic serum concentrations of alpha-carotene, beta-carotene, beta-cryptoxanthin, lycopene, lutein/zeaxanthin, retinol, alpha-tocopherol, gamma-tocopherol, and vitamin C were determined on 191 cases and 570 matched controls within a cohort of 18,244 middle-aged or older men in Shanghai, China, with a follow-up of 12 years. High serum levels of alpha carotene, beta-carotene, and lycopene were significantly associated with reduced risk of developing gastric cancer (all Ps for trend /=3 drinks of alcohol per day; the odds ratios (95% confidence intervals) for the second, third, and fourth quartile categories were 0.69 (0.28-1.70), 0.36 (0.14-0.94), and 0.39 (0.15 0.98), respectively, compared with the lowest quartile of vitamin C (P for trend = 0.02). There were no statistically significant relationships of serum levels of beta-cryptoxanthin, lutein/zeaxanthin, retinol, alpha-tocopherol, and gamma tocopherol with gastric cancer risk. The present study implicates that dietary carotenes, lycopene, and vitamin C are potential chemopreventive agents for gastric cancer in humans. PMID- 15533907 TI - Serum oxidized low-density lipoprotein levels and risk of colorectal cancer: a case-control study nested in the Japan Collaborative Cohort Study. AB - Oxidative stress plays an important role in carcinogenesis, but few epidemiologic studies have examined associations with risk of colorectal cancer. Relationships between serum levels of oxidized low-density lipoprotein (oxLDL) and oxLDL antibody (oLAB) and colorectal cancer risk were investigated in a case-control study nested in the Japan Collaborative Cohort Study for Evaluation of Cancer Risk. Serum samples and lifestyle information were collected at baseline from 39,242 men and women between 1988 and 1990. Of these, 161 incidents and deaths from colorectal cancer were identified through 1999, and 395 controls were matched for gender, age, and study area. Measurements were taken of serum oxLDL levels in 119 cases and 316 controls and serum oLAB levels in 153 cases and 376 controls. Odds ratios (95% confidence intervals) across quartiles, adjusted for confounding factors, were 1.55 (0.70-3.46), 1.90 (0.84-4.28), and 3.65 (1.50 8.92) for oxLDL (P(trend) = 0.004) and 0.98 (0.54-1.80), 0.75 (0.39-1.48), and 1.68 (0.90-3.13) for oLAB (P(trend) = 0.140). Further adjustment for serum total cholesterol and alpha-tocopherol did not materially change these associations. Odds ratio (95% confidence interval) of the highest quartile of serum oxLDL compared with the lowest quartile was 3.40 (1.09-10.58; P(trend) = 0.045). Analyses restricted to colon cancer cases and corresponding controls yielded similar relationships between serum oxLDL and oLAB levels and risk. In conclusion, higher levels of serum oxLDL may increase risk of colorectal cancer. PMID- 15533908 TI - Poly (AT) polymorphism in intron 11 of the XPC DNA repair gene enhances the risk of lung cancer. AB - Reduced DNA repair capacity due to inherited polymorphisms may increase the susceptibility to smoking-related cancers. In this report, we investigate the relationship between xeroderma pigmentosum complementary group C poly (AT) insertion/deletion polymorphism (XPC-PAT) of the XPC gene and lung cancer risk in a hospital-based case-control study of 359 newly diagnosed lung cancer patients and 375 control subjects matched on age, sex, and catchment area. The XPC genotype was determined by PCR-RFLP, and the results were analyzed using logistic regression, adjusting for relevant covariates. We found that the frequency of the PAT+/+ genotype was higher in the cases (20.6%) than in the controls (14.1%; P = 0.057) and that the PAT+/+ subjects were at significantly increased risk for lung cancer [adjusted odds ratio (OR), 1.60; 95% confidence interval (95% CI), 1.01 2.55]. Stratified analysis revealed that the risk was higher in former smokers (OR, 2.15; 95% CI, 1.07-4.31) and older people (OR, 2.76; 95% CI, 1.02-7.51), although this probably occurs due to 63.4% of cases older than 73 years being ex smokers. When stratified by histologic type, the variant genotype was associated with statistically significant increased risk for squamous cell carcinoma (OR, 1.93; 95% CI, 1.06-3.51). In conclusion, our findings support the hypothesis that PAT and intron 11 C/A XPC polymorphisms are linked in the Spanish population and may contribute to the risk of developing lung cancer probably due to a higher frequency of deletion of exon 12 and reduced DNA repair capacity of the XPC protein. PMID- 15533909 TI - BRCA1 and BRCA2 mutations in a study of African American breast cancer patients. AB - The spectrum of mutations in BRCA1 and BRCA2 among African Americans has not been well characterized because most studies to date have been done in Caucasian families. According to Myriad Genetic Laboratories, Inc., only approximately 3% of individuals undergoing BRCA1/BRCA2 testing reported African American ancestry. Data from previous studies show that among African American women a greater proportion of breast cancer cases are diagnosed at age <45 years in comparison with Caucasians. Because breast cancer occurring at a young age is one of the hallmarks of high penetrance genes, the prevalence, spectrum, and effects of BRCA1/BRCA2 mutations may differ substantially between African Americans and Caucasians, and further investigation is warranted. We conducted a hospital-based study of African American breast cancer patients with early age at diagnosis (A). Our results support previous data in demonstrating that (a) the spectrum of mutations among African Americans is unique, (b) family history of breast cancer is an important predictor of hereditary cancer susceptibility among African Americans, and (c) empirical data may be useful in estimating mutation risk among African Americans. PMID- 15533910 TI - Smoking topography predicts abstinence following treatment with nicotine replacement therapy. AB - OBJECTIVE: Smoking topography refers to how a person smokes a cigarette and includes measures of the number of puffs and puff volume, duration, and velocity. This study examined the relationship between smoking topography and abstinence from cigarettes following nicotine replacement therapy. A secondary objective was to determine the relationship of smoking topography to carbon monoxide (CO) exposure. METHOD: Participants (n = 113) smoked one of their preferred brands of cigarette through a smoking topography device prior to participating in an open label trial of transdermal nicotine versus nicotine nasal spray. A subset of participants (n = 50) provided breath CO samples prior to and following smoking the cigarette. RESULTS: Mean V(max) [odds ratio (OR), 1.12; 95% confidence interval (95% CI), 1.02-1.24; P = 0.02], mean puff volume (OR, 0.95; 95% CI, 0.91 0.98; P = 0.01), mean interpuff interval (OR, 1.06; 95% CI, 1.00-1.11; P = 0.03), and cigarette type (full flavor versus light/ultralight; OR, 0.35; 95% CI, 0.14 0.89; P = 0.03) were significant predictors of abstinence in a model controlling for treatment group and nicotine dependence. Controlling for time since last cigarette and initial CO level, mean puff velocity (beta = 0.171; P = 0.01) was the only significant predictor of CO boost. CONCLUSION: These results suggest that smoking topography may be useful to predict abstinence after using nicotine replacement therapy and to assess harm from smoking. PMID- 15533911 TI - p53 codon 72 polymorphic variants, loss of allele-specific transcription, and human papilloma virus 16 and/or 18 E6 messenger RNA expression in squamous cell carcinomas of the head and neck. AB - A polymorphism at codon 72 of the human tumor suppressor p53 determines translation into either arginine or proline. Yet, the impact of this amino acid variability on the risk to develop malignant tumors, particularly carcinomas associated with human papilloma virus (HPV) infections, remains unresolved because of contradictory results. To address a potential correlation between the different genotypes and the manifestation of squamous cell carcinomas of the head and neck (SCCHN), we determined the p53 codon 72 in 193 healthy subjects and 122 unselected SCCHN with known HPV status. Furthermore, loss of allele-specific transcription was analyzed in p53 codon 72 heterozygous (Arg/Pro) SCCHN and correlated with HPV 16 and/or 18 E6 transcript expression. We found a moderately increased risk (odds ratio, 1.86; 95% confidence interval, 1.0-3.3) for individuals with germ line heterozygosity to develop SCC of the pharynx. On the other hand, p53 codon 72 polymorphic variants, most notably the Arg/Arg genotype, showed no association with the presence of HPV 16 and/or 18 E6 transcript. Moreover, there was no evidence for HPV-driven selection in SCCHN with allele specific loss of transcription. Our data suggest that the p53 codon 72 polymorphism has a minor impact on the development of SCCHN. PMID- 15533912 TI - Body mass index and risk of leukemia in older women. AB - Overweight [body mass index (BMI) 25.0-29.9 kg/m(2)] and obesity (BMI >/=30 kg/m(2)) are risk factors for several malignancies. The Iowa Women's Health Study was examined to determine whether increased BMI was associated with leukemia development. Over 40,000 Iowa women (ages 55-69 years) completed a self administered lifestyle and health questionnaire in 1986 that included current height and weight. Two hundred women developed leukemia during the period 1986 to 2001 including 74 acute myelogenous leukemia (AML) and 88 chronic lymphocytic leukemia. The risk of AML was increased among women who reported being overweight or obese (relative risk, 1.9; 95% confidence interval, 1.0-3.4; relative risk, 2.4; 95% confidence interval, 1.3-4.5; P(trend) = 0.006) compared with women of normal weight. There was little evidence of a positive association for chronic lymphocytic leukemia (P(trend) = 0.6). Given the prevalence of overweight and obesity in the United States, the population attributable risk of AML due to obesity could approach 30%. PMID- 15533913 TI - Occupational exposure to immunologically active agents and risk for lymphoma. AB - OBJECTIVES: We evaluated whether occupational exposure to high molecular weight agents that are associated with asthma and that act predominantly through an immunoglobulin E (IgE)-dependent hypersensitivity mechanism is also associated with risk for specific lymphoma types. METHODS: The Spanish lymphoma case-control study includes 519 newly diagnosed cases of lymphoid neoplasms and 554 hospital controls ages 20 to 80 years. Lymphomas were histologically or cytologically confirmed and classified according to the WHO classification. Lifetime occupational exposure to seven high molecular weight agents such as latex was evaluated through an asthma-specific job-exposure matrix and validated by an industrial hygienist. Odds ratios (OR) and 95% confidence intervals (95% CI) were adjusted for age, sex, hospital, and occupational exposures. RESULTS: Subjects exposed to high molecular weight agents had an increased risk for Hodgkin's lymphoma (OR, 2.27; 95% CI, 0.93-5.50), particularly nodular sclerosis (OR, 3.22; 95% CI, 1.14-9.09). ORs did not increase with duration of exposure to these agents. Exposure to high molecular weight agents was not associated with risk of other B-cell lymphomas; for most specific subcategories of B-cell lymphoma, ORs were below unity. A slightly increased OR (95% CI) was seen for mycosis fungoides [1.60 (0.53-4.84)], although overall there was no increased risk for T-cell lymphomas. CONCLUSIONS: Exposure to immunologically active agents among clinically immunocompetent subjects was associated with risk for Hodgkin's lymphoma. PMID- 15533914 TI - No association between polyomaviruses and primary central nervous system lymphomas of HIV-seronegative and HIV-positive patients. PMID- 15533915 TI - No association between GPX1 Pro198Leu and breast cancer risk. PMID- 15533916 TI - Wine consumption and epithelial ovarian cancer. PMID- 15533917 TI - Functional characterization of mammalian inositol 1,4,5-trisphosphate receptor isoforms. AB - Inositol 1,4,5-trisphosphate receptors (InsP3R) play a key role in intracellular calcium (Ca2+) signaling. Three mammalian InsP3R isoforms--InsP3R type 1 (InsP3R1), InsP3R type 2 (InsP3R2), and InsP3R type 3 (InsP3R3) are expressed in mammals, but the functional differences between the three mammalian InsP3R isoforms are poorly understood. Here we compared single-channel behavior of the recombinant rat InsP3R1, InsP3R2, and InsP3R3 expressed in Sf9 cells, reconstituted into planar lipid bilayers and recorded with 50 mM Ba2+ as a current carrier. We found that: 1), for all three mammalian InsP3R isoforms the size of the unitary current is 1.9 pA and single-channel conductance is 74-80 pS; 2), in optimal recording conditions the maximal single-channel open probability for all three mammalian InsP3R isoforms is in the range 30-40%; 3), in optimal recording conditions the mean open dwell time for all three mammalian InsP3R isoforms is 7-8 ms, the mean closed dwell time is approximately 10 ms; 4), InsP3R2 has the highest apparent affinity for InsP(3) (0.10 microM), followed by InsP3R1 (0.27 microM), and then by InsP3R3 (0.40 microM); 5), InsP3R1 has a high affinity (0.13 mM) ATP modulatory site, InsP3R2 gating is ATP independent, and InsP3R3 has a low-affinity (2 mM) ATP modulatory site; 6), ATP modulates InsP3R1 gating in a noncooperative manner (n(Hill) = 1.3); 7), ATP modulates InsP3R3 gating in a highly cooperative manner (n(Hill) = 4.1). Obtained results provide novel information about functional properties of mammalian InsP3R isoforms. PMID- 15533918 TI - Association of the Igamma and Idelta charge movement with calcium release in frog skeletal muscle. AB - Charge movement and calcium transient were measured simultaneously in stretched frog cut twitch fibers under voltage clamp, with the internal solution containing 20 mM EGTA plus added calcium and antipyrylazo III. When the nominal free [Ca2+]i was 10 nM, the shape of the broad I(gamma) hump in the ON segments of charge movement traces remained invariant when the calcium release rate was greatly diminished. When the nominal free [Ca2+]i was 50 nM, which was close to the physiological level, the I(gamma) humps were accelerated and a slow calcium dependent I(delta) component (or state) was generated. The peak of ON I(delta) synchronized perfectly with the peak of the calcium release rate whereas the slow decay of ON I(delta) followed the same time course as the decay of calcium release rate. Suppression of calcium release by TMB-8 reduced the amount of Q(delta) concomitantly but not completely, and the effects were partially reversible. The same simultaneous suppression effects were achieved by depleting the sarcoplasmic reticulum calcium store with repetitive stimulation. The results suggest that the mobility of Q(delta) needs to be primed by a physiological level of resting myoplasmic Ca2+. Once the priming is completed, more I(delta) is mobilized by the released Ca2+ during depolarization. PMID- 15533919 TI - Measuring single cardiac myocyte contractile force via moving a magnetic bead. AB - One of the biggest problems of heart failure is the heart's inability to effectively pump blood to meet the body's demands, which may be caused by disease induced alterations in contraction properties (such as contractile force and Young's modulus). Thus, it is very important to measure contractile properties at single cardiac myocyte level that can lay the foundation for quantitatively understanding the mechanism of heart failure and understanding molecular alterations in diseased heart cells. In this article, we report a novel single cardiac myocyte contractile force measurement technique based on moving a magnetic bead. The measuring system is mainly composed of 1), a high-power inverted microscope with video output and edge detection; and 2), a moving magnetic bead based magnetic force loading module. The main measurement procedures are as follows: 1), record maximal displacement of single cardiac myocyte during contraction; 2), attach a magnetic bead on one end of the myocyte that will move with myocyte during the contraction; 3), repeat step 1 and record contraction processes under different magnitudes of magnetic force loading by adjusting the magnetic field applied on the magnetic bead; and 4), derive the myocyte contractile force base on the maximal displacement of cell contraction and magnetic loading force. The major advantages of this unique approach are: 1), measuring the force without direct connections to the cell specimen (i.e., "remote sensing", a noninvasive/minimally invasive approach); 2), high sensitivity and large dynamic range (force measurement range: from pico Newton to micro Newton); 3), a convenient and cost-effective approach; and 4), more importantly, it can be used to study the contractile properties of heart cells under different levels of external loading forces by adjusting the magnitude of applied magnetic field, which is very important for studying disease induced alterations in contraction properties. Experimental results demonstrated the feasibility of proposed approach. PMID- 15533920 TI - Nanoscale features of fibronectin fibrillogenesis depend on protein-substrate interaction and cytoskeleton structure. AB - Cell-reorganized fibronectin layers on polymer films providing a gradation of the binding strength between protein and substrate were analyzed by combined fluorescence and scanning force microscopy. The nanoscale fibronectin patterns exhibited paired parallel fibrils with characteristic spacings of 156, 233, 304, and 373 nm. These spacings depend on the interaction of fibronectin with the substrate: at enhanced fibronectin-substrate anchorage the cells form larger stress fibers, which are assembled by alpha-actinin cross-linked pairs of actin filaments subunits at the focal adhesions. A ubiquitous repeating unit of approximately 71 nm was found within these characteristic distances. We conclude that the dimensions of the actin stress fibers reflect the binding strength of fibronectin to the polymer substrate and act--in turn--as a template for the reorganization of fibronectin into surface-bound nanofibrils with characteristic spacings. This explanation was confirmed by data showing the alpha actinin/fibronectin colocalization. PMID- 15533921 TI - Conformational states of the Rapana thomasiana hemocyanin and its substructures studied by dynamic light scattering and time-resolved fluorescence spectroscopy. AB - Hemocyanins are dioxygen-transporting proteins freely dissolved in the hemolymph of mollusks and arthropods. Dynamic light scattering and time-resolved fluorescence measurements show that the oxygenated and apo-forms of the Rapana thomasiana hemocyanin, its structural subunits RtH1 and RtH2, and those of the functional unit RtH2e, exist in different conformations. The oxygenated respiratory proteins are less compact and more asymmetric than the respective apo forms. Different conformational states were also observed for the R. thomasiana hemocyanin in the absence and presence of an allosteric regulator. The results are in agreement with a molecular mechanism for cooperative dioxygen binding in molluscan hemocyanins including transfer of conformational changes from one functional unit to another. PMID- 15533922 TI - Interpreting second-harmonic generation images of collagen I fibrils. AB - Fibrillar collagen, being highly noncentrosymmetric, possesses a tremendous nonlinear susceptibility. As a result, second-harmonic generation (SHG) microscopy of collagen produces extremely bright and robust signals, providing an invaluable tool for imaging tissue structure with submicron resolution. Here we discuss fundamental principles governing SHG phase matching with the tightly focusing optics used in microscopy. Their application to collagen imaging yields several biophysical features characteristic of native collagen structure: SHG radiates from the shell of a collagen fibril, rather than from its bulk. This SHG shell may correspond to the supporting element of the fibril. Physiologically relevant changes in solution ionic strength alter the ratio of forward-to backward propagating SHG, implying a resulting change in the SHG shell thickness. Fibrillogenesis can be resolved in immature tissue by directly imaging backward propagating SHG. Such findings are crucial to the design and development of forthcoming diagnostic and research tools. PMID- 15533923 TI - Mechanics of transient platelet adhesion to von Willebrand factor under flow. AB - A primary and critical step in platelet attachment to injured vascular endothelium is the formation of reversible tether bonds between the platelet glycoprotein receptor Ibalpha and the A1 domain of surface-bound von Willebrand factor (vWF). Due to the platelet's unique ellipsoidal shape, the force mechanics involved in its tether bond formation differs significantly from that of leukocytes and other spherical cells. We have investigated the mechanics of platelet tethering to surface-immobilized vWF-A1 under hydrodynamic shear flow. A computer algorithm was used to analyze digitized images recorded during flow chamber experiments and track the microscale motions of platelets before, during, and after contact with the surface. An analytical two-dimensional model was developed to calculate the motion of a tethered platelet on a reactive surface in linear shear flow. Through comparison of the theoretical solution with experimental observations, we show that attachment of platelets occurs only in orientations that are predicted to result in compression along the length of the platelet and therefore on the bond being formed. These results suggest that hydrodynamic compressive forces may play an important role in initiating tether bond formation. PMID- 15533924 TI - Molecular dynamics simulations of discoidal bilayers assembled from truncated human lipoproteins. AB - Human apolipoprotein A-1 (apo A-1) is the major protein component of high-density lipoproteins. The apo A-1 lipid-binding domain was used as a template for the synthesis of amphipathic helical proteins termed membrane scaffold proteins, employed to self-assemble soluble monodisperse discoidal particles called Nanodiscs. In these particles, membrane scaffold proteins surround a lipid bilayer in a belt-like fashion forming bilayer disks of discrete size and composition. Here we investigate the structure of Nanodiscs through molecular dynamics simulations in which Nanodiscs were built from scaffold proteins of various lengths. The simulations showed planar or deformed Nanodiscs depending on optimal length and alignment of the scaffold proteins. Based on mean surface area per lipid calculations, comparison of small-angle x-ray scattering curves, and the relatively planar shape of Nanodiscs made from truncated scaffold proteins, one can conclude that the first 17 to 18 residues of the 200-residue apo A-1 lipid-binding domain are not involved in formation of the protein "belts" surrounding the lipid bilayer. To determine whether the addition of an integral membrane protein has an effect on the overall structure of a Nanodisc, bacteriorhodopsin was embedded into a Nanodisc and simulated using molecular dynamics, revealing a planar disk with a slightly rectangular shape. PMID- 15533925 TI - Experimental validation of molecular dynamics simulations of lipid bilayers: a new approach. AB - A novel protocol has been developed for comparing the structural properties of lipid bilayers determined by simulation with those determined by diffraction experiments, which makes it possible to test critically the ability of molecular dynamics simulations to reproduce experimental data. This model-independent method consists of analyzing data from molecular dynamics bilayer simulations in the same way as experimental data by determining the structure factors of the system and, via Fourier reconstruction, the overall transbilayer scattering density profiles. Multi-nanosecond molecular dynamics simulations of a dioleoylphosphatidylcholine bilayer at 66% RH (5.4 waters/lipid) were performed in the constant pressure and temperature ensemble using the united-atom GROMACS and the all-atom CHARMM22/27 force fields with the GROMACS and NAMD software packages, respectively. The quality of the simulated bilayer structures was evaluated by comparing simulation with experimental results for bilayer thickness, area/lipid, individual molecular-component distributions, continuous and discrete structure factors, and overall scattering-density profiles. Neither the GROMACS nor the CHARMM22/27 simulations reproduced experimental data within experimental error. The widths of the simulated terminal methyl distributions showed a particularly strong disagreement with the experimentally observed distributions. A comparison of the older CHARMM22 with the newer CHARMM27 force fields shows that significant progress is being made in the development of atomic force fields for describing lipid bilayer systems empirically. PMID- 15533927 TI - Correlation of the O-intermediate rate with the pKa of Asp-75 in the dark, the counterion of the Schiff base of Pharaonis phoborhodopsin (sensory rhodopsin II). AB - Pharaonis phoborhodopsin (ppR), also called pharaonis sensory rhodopsin II, NpSRII, is a photoreceptor of negative phototaxis in Natronomonas (Natronobacterium) pharaonis. The photocycle rate of ppR is slow compared to that of bacteriorhodopsin, despite the similarity in their x-ray structures. The decreased rate of the photocycle of ppR is a result of the longer lifetime of later photo-intermediates such as M- (ppR(M)) and O-intermediates (ppR(O)). In this study, mutants were prepared in which mutated residues were located on the extracellular surface (P182, P183, and V194) and near the Schiff base (T204) including single, triple (P182S/P183E/V194T), and quadruple mutants. The decay of ppR(O) of the triple mutant was accelerated approximately 20-times from 690 ms for the wild-type to 36 ms. Additional mutation resulting in a triple mutant at the 204th position such as T204C or T204S further decreased the decay half-time to 6.6 or 8 ms, almost equal to that of bacteriorhodopsin. The decay half-times of the ppR(O) of mutants (11 species) and those of the wild-type were well correlated with the pK(a) value of Asp-75 in the dark for the respective mutants as spectroscopically estimated, although there are some exceptions. The implications of these observations are discussed in detail. PMID- 15533926 TI - Folding Trp-cage to NMR resolution native structure using a coarse-grained protein model. AB - We develop a coarse-grained protein model with a simplified amino acid interaction potential. Using this model, we perform discrete molecular dynamics folding simulations of a small 20-residue protein--Trp-cage--from a fully extended conformation. We demonstrate the ability of the Trp-cage model to consistently reach conformations within 2-angstroms backbone root-mean-square distance from the corresponding NMR structures. The minimum root-mean-square distance of Trp-cage conformations in simulations can be <1 angstroms. Our findings suggest that, at least in the case of Trp-cage, a detailed all-atom protein model with a molecular mechanics force field is not necessary to reach the native state of a protein. Our results also suggest that the success of folding Trp-cage in our simulations and in the reported all-atom molecular mechanics simulation studies may be mainly due to the special stabilizing features specific to this miniprotein. PMID- 15533928 TI - Static and dynamic errors in particle tracking microrheology. AB - Particle tracking techniques are often used to assess the local mechanical properties of cells and biological fluids. The extracted trajectories are exploited to compute the mean-squared displacement that characterizes the dynamics of the probe particles. Limited spatial resolution and statistical uncertainty are the limiting factors that alter the accuracy of the mean-squared displacement estimation. We precisely quantified the effect of localization errors in the determination of the mean-squared displacement by separating the sources of these errors into two separate contributions. A "static error" arises in the position measurements of immobilized particles. A "dynamic error" comes from the particle motion during the finite exposure time that is required for visualization. We calculated the propagation of these errors on the mean-squared displacement. We examined the impact of our error analysis on theoretical model fluids used in biorheology. These theoretical predictions were verified for purely viscous fluids using simulations and a multiple-particle tracking technique performed with video microscopy. We showed that the static contribution can be confidently corrected in dynamics studies by using static experiments performed at a similar noise-to-signal ratio. This groundwork allowed us to achieve higher resolution in the mean-squared displacement, and thus to increase the accuracy of microrheology studies. PMID- 15533929 TI - Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. AB - The consumption of food products containing high amounts of flavonoids has been reported to lower the risk of various cancers. The mechanisms underlying the cancer-protective effects of these naturally occurring polyphenolic compounds, however, remain elusive. Based on our previous finding that the cytotoxic effect of the flavanol epigallocatechin-3-gallate on prostate cancer cells correlates with its ability to inhibit fatty acid synthase (FAS, a key lipogenic enzyme overexpressed in many human cancers), we examined the anti-lipogenic effects of a panel of 18 naturally occurring polyphenolic compounds. In addition to epigallocatechin-3-gallate, five other flavonoids, more particularly luteolin, quercetin, kaempferol, apigenin, and taxifolin, also markedly inhibited cancer cell lipogenesis. Interestingly, in both prostate and breast cancer cells, a remarkable dose-response parallelism was observed between flavonoid-induced inhibition of fatty acid synthesis, inhibition of cell growth, and induction of apoptosis. In support for a role of fatty acid synthesis in these effects, the addition of exogenous palmitate, the end product of FAS, markedly suppressed the cytotoxic effects of flavonoids. Taken together, these findings indicate that the potential of flavonoids to induce apoptosis in cancer cells is strongly associated with their FAS inhibitory properties, thereby providing a new mechanism by which polyphenolic compounds may exert their cancer-preventive and antineoplastic effects. PMID- 15533930 TI - Targeting of enteropathogenic Escherichia coli EspF to host mitochondria is essential for bacterial pathogenesis: critical role of the 16th leucine residue in EspF. AB - The attachment of enteropathogenic Escherichia coli (EPEC) to host cells and the induction of attaching and effacing (A/E) lesions are prominent pathogenic features. EPEC infection also leads to host cell death and damage to the intestinal mucosa, which is partly dependent upon EspF, one of the effectors. In this study, we demonstrate that EspF is a mitochondrial import protein with a functional mitochondrial targeting signal (MTS), because EspF activity for importing into the mitochondria was abrogated by MTS deletion mutants. Substitution of the 16th leucine with glutamic acid (EspF(L16E)) completely abolished EspF activity. Infection of HeLa cells with wild type but not the espF mutant (DeltaespF) decreased mitochondrial membrane potential (DeltaPsi(m)), leading to cell death. The DeltaPsi(m) decrease and cell death were restored in cells infected with DeltaespF/pEspF but not DeltaespF/pEspF(L16E), suggesting that the 16th leucine in the MTS is a critical amino acid for EspF function. To demonstrate the impact of EspF in vivo, we exploited Citrobacter rodentium by infecting C3H/HeJ mice with DeltaespF(CR), DeltaespF(CR)/pEspF(CR), or DeltaespF(CR)/pEspF(L16E)(CR). These results indicate that EspF activity contributes to bacterial pathogenesis, as judged by murine lethality and intestinal histopathology, and promotion of bacterial colonization of the intestinal mucosa. PMID- 15533931 TI - Assignment of the binding site for haptoglobin on apolipoprotein A-I. AB - Haptoglobin (Hpt) was previously found to bind the high density lipoprotein (HDL) apolipoprotein A-I (ApoA-I) and able to inhibit the ApoA-I-dependent activity of the enzyme lecithin:cholesterol acyltransferase (LCAT), which plays a major role in the reverse cholesterol transport. The ApoA-I structure was analyzed to detect the site bound by Hpt. ApoA-I was treated by cyanogen bromide or hydroxylamine; the resulting fragments, separated by electrophoresis or gel filtration, were tested by Western blotting or enzyme-linked immunosorbent assay for their ability to bind Hpt. The ApoA-I sequence from Glu113 to Asn184 harbored the binding site for Hpt. Biotinylated peptides were synthesized overlapping such a sequence, and their Hpt binding activity was determined by avidin-linked peroxidase. The highest activity was exhibited by the peptide P2a, containing the ApoA-I sequence from Leu141 to Ala164. Such a sequence contains an ApoA-I domain required for binding cells, promoting cholesterol efflux, and stimulating LCAT. The peptide P2a effectively prevented both binding of Hpt to HDL-coated plastic wells and Hpt dependent inhibition of LCAT, measured by anti-Hpt antibodies and cholesterol esterification activity, respectively. The enzyme activity was not influenced, in the absence of Hpt, by P2a. Differently from ApoA-I or HDL, the peptide did not compete with hemoglobin for Hpt binding in enzyme-linked immunosorbent assay experiments. The results suggest that Hpt might mask the ApoA-I domain required for LCAT stimulation, thus impairing the HDL function. Synthetic peptides, able to displace Hpt from ApoA-I without altering its property of binding hemoglobin, might be used for treatment of diseases associated with defective LCAT function. PMID- 15533932 TI - HDQ (1-hydroxy-2-dodecyl-4(1H)quinolone), a high affinity inhibitor for mitochondrial alternative NADH dehydrogenase: evidence for a ping-pong mechanism. AB - Alternative NADH dehydrogenases (NADH:ubiquinone oxidoreductases) are single subunit respiratory chain enzymes found in plant and fungal mitochondria and in many bacteria. It is unclear how these peripheral membrane proteins interact with their hydrophobic substrate ubiquinone. Known inhibitors of alternative NADH dehydrogenases bind with rather low affinities. We have identified 1-hydroxy-2 dodecyl-4(1H)quinolone as a high affinity inhibitor of alternative NADH dehydrogenase from Yarrowia lipolytica. Using this compound, we have analyzed the bisubstrate and inhibition kinetics for NADH and decylubiquinone. We found that the kinetics of alternative NADH dehydrogenase follow a ping-pong mechanism. This suggests that NADH and the ubiquinone headgroup interact with the same binding pocket in an alternating fashion. PMID- 15533933 TI - Ataxia telangiectasia mutated (ATM) and ATM and Rad3-related protein exhibit selective target specificities in response to different forms of DNA damage. AB - The ataxia telangiectasia mutated (ATM) and ATR (ATM and Rad3-related) protein kinases exert cell cycle delay, in part, by phosphorylating Checkpoint kinase (Chk) 1, Chk2, and p53. It is well established that ATR is activated following UV light-induced DNA damage such as pyrimidine dimers and the 6-(1,2)-dihydro-2-oxo 4-pyrimidinyl-5-methyl-2,4-(1H,3H)-pyrimidinediones, whereas ATM is activated in response to double strand DNA breaks. Here we clarify the activation of these kinases in cells exposed to IR, UV, and hyperoxia, a condition of chronic oxidative stress resulting in clastogenic DNA damage. Phosphorylation on Chk1(Ser 345), Chk2(Thr-68), and p53(Ser-15) following oxidative damage by IR involved both ATM and ATR. In response to ultraviolet radiation-induced stalled replication forks, phosphorylation on Chk1 and p53 required ATR, whereas Chk2 required ATM. Cells exposed to hyperoxia exhibited growth delay in G1, S, and G2 that was disrupted by wortmannin. Consistent with ATM or ATR activation, hyperoxia induced wortmannin-sensitive phosphorylation of Chk1, Chk2, and p53. By using ATM- and ATR-defective cells, phosphorylation on Chk1, Chk2, and p53 was found to be ATM-dependent, whereas ATR also contributed to Chk1 phosphorylation. These data reveal activated ATM and ATR exhibit selective substrate specificity in response to different genotoxic agents. PMID- 15533934 TI - DNA topoisomerase II poison TAS-103 transactivates GC-box-dependent transcription via acetylation of Sp1. AB - Drug-induced modifications of transcription factors play important roles in both apoptosis and survival signaling. The data presented here show that the DNA topoisomerase II poison TAS-103 transactivated the SV40 promoter in a GC-box dependent manner and induced Sp1 acetylation in cells expressing p300. This activity was not observed in cells lacking p300. TAS-103 treatment also enhanced the p300 content of the nucleus and the interaction of p300 with Sp1. Cellular susceptibility to TAS-103 was correlated with p300 expression but not with topoisomerase II expression. Furthermore, the presence of p300 significantly sensitized cancer cells to TAS-103 but not to cisplatin. Taken together, these findings demonstrate novel genomic responses to anticancer agents that modulate Sp1 acetylation and Sp1-dependent transcription in an apoptotic pathway. PMID- 15533935 TI - Association of NASP with HSP90 in mouse spermatogenic cells: stimulation of ATPase activity and transport of linker histones into nuclei. AB - NASP (nuclear autoantigenic sperm protein) is a linker histone-binding protein found in all dividing cells that is regulated by the cell cycle (Richardson, R. T., Batova, I. N., Widgren, E. E., Zheng, L. X., Whitfield, M., Marzluff, W. F., and O'Rand, M. G. (2000) J. Biol. Chem. 275, 30378-30386), and in the nucleus linker histones not bound to DNA are bound to NASP (Alekseev, O. M., Bencic, D. C., Richardson R. T., Widgren E. E., and O'Rand, M. G. (2003) J. Biol. Chem. 278, 8846-8852). In mouse spermatogenic cells tNASP binds the testis-specific linker histone H1t. Utilizing a cross-linker, 3,3'-dithiobissulfosuccinimidyl propionate, and mass spectrometry, we have identified HSP90 as a testis/embryo form of NASP (tNASP)-binding partner. In vitro assays demonstrate that the association of tNASP with HSP90 stimulated the ATPase activity of HSP90 and increased the binding of H1t to tNASP. HSP90 and tNASP are present in both nuclear and cytoplasmic fractions of mouse spermatogenic cells; however, HSP90 bound to NASP only in the cytoplasm. In vitro nuclear import assays on permeabilized HeLa cells demonstrate that tNASP, in the absence of any other cytoplasmic factors, transports linker histones into the nucleus in an energy and nuclear localization signal-dependent manner. Consequently we hypothesize that in the cytoplasm linker histones are bound to a complex containing NASP and HSP90 whose ATPase activity is stimulated by binding NASP. NASP-H1 is subsequently released from the complex and translocates to the nucleus where the H1 is released for binding to the DNA. PMID- 15533936 TI - Enhanced dephosphorylation of cAMP-dependent protein kinase by oxidation and thiol modification. AB - The catalytic subunit of cAMP-dependent protein kinase (PKA) is phosphorylated at threonine 197 and serine 338. Phosphorylation of threonine 197, located in the activation loop, is required for coordinating the active site conformation and optimal enzymatic activity. However, this phosphorylation has not been widely appreciated as a regulatory site because of the apparent constitutive nature of the phosphorylation and the general resistance of the kinase to phosphatase treatment. We demonstrate here that the observed resistance of the catalytic subunit to dephosphorylation is due, in part, to the presence of the highly nucleophilic cysteine 199 located proximal to the phosphate on threonine 197. Experiments performed in vitro demonstrated that mutation (cysteine 199 to alanine), oxidation, such as by glutathionylation or internal disulfide bond formation, or alkylation of the C-subunit enhanced its ability to be dephosphorylated. Furthermore, rephosphorylation of reduced C-subunit by PDK1 created a cycle whereby the inactive kinase could be reactivated. To demonstrate that thiol modification of PKA can lead to enhanced dephosphorylation in vivo, PC12 cells were treated with N-ethylmaleimide (NEM). Such treatment resulted in complete PKA inactivation and dephosphorylation of threonine 197. This effect of NEM was contingent upon prior treatment of the cells with PKA activators, demonstrating the resistance of the holoenzyme to thiol alkylation-mediated dephosphorylation. Our results also demonstrated that NEM treatment of PC12 cells enhanced the dephosphorylation of the protein kinase Calpha activation loop, suggesting a common mechanism of regulation among members of the AGC family of kinases. PMID- 15533937 TI - CAG repeats containing CAA interruptions form branched hairpin structures in spinocerebellar ataxia type 2 transcripts. AB - Spinocerebellar ataxia type 2 (SCA2), one of the hereditary human neurodegenerative disorders, is caused by the expansion of the CAG tandem repeats in the translated sequence of the SCA2 gene. In a normal population the CAG repeat is polymorphic not only in length but also in the number and localization of its CAA interruptions. The aim of this study was to determine the structure of the repeat region in the normal and mutant SCA2 transcripts and to reveal the structural basis of its normal function and dysfunction. We show here that the properties of the CAA interruptions are major determinants of the CAG repeat folding in the normal SCA2 transcripts. We also show that the uninterrupted repeats in mutant transcripts form slippery hairpins, whose length is further reduced by the base pairing of the repeat portion with a specific flanking sequence. The structural organization of the repeat interruption systems present in other human transcripts, such as SCA1, TBP, FOXP2, and MAML2, are also discussed. PMID- 15533938 TI - Pivotal molecular determinants of peptidic and collagen triple helicase activities reside in the S3' subsite of matrix metalloproteinase 8 (MMP-8): the role of hydrogen bonding potential of ASN188 and TYR189 and the connecting cis bond. AB - The mechanism of triple helical collagen unwinding and cleavage by collagenases in the matrix metalloproteinase (MMP) family is complex and remains enigmatic. Recent reports show that triple helicase activity is initiated by the hemopexin C domain of membrane type 1-MMP, whereas catalytically inactive full-length interstitial collagenase (MMP-1) exhibits full triple helicase functionality pointing to active site determinants that are needed to complete the triple helicase mechanism. In MMP-8, the neutrophil collagenase, a conserved Gly at the S(3)' substrate specificity subsite is replaced by Asn(188) that forms a highly unusual cis bond with Tyr(189), a conserved active site residue in the collagenases. Only in MMP-1 is the S(3)' Gly also replaced, and there too a cis configured Glu-Tyr occurs. Thus, this high energy peptide bond coupled to the canonical Tyr may be important in the collagenolytic process. In a systematic mutagenesis investigation of the MMP-8 S(3)' subsite we found that introducing an S(3)' Gly(188) into MMP-8 reduced collagenolytic efficiency by approximately 30% with a corresponding reduction in cleavage of a synthetic peptide fluorescence resonance energy transfer substrate analogue of the alpha2(I) collagen chain cleavage site. The substitution of Asn(188) to Leu, a hydrophobic residue of similar size to the highly polar Asn and designed to retain the cis bond, revealed the importance of hydrogen bonding to bound substrate with both collagenolytic and peptidic activities reduced approximately 3-fold. In contrast, the specificity for type I collagen of the mutant Y189F dropped 3-fold without any significant alteration in general peptidase activity. Therefore, S(3)' and in particular the hydrogen bonding potential of Tyr(189) is a specific molecular determinant for MMP-8 triple helicase activity. The cis bond connection to Asn(188) juxtaposes these two side chains for closely spaced hydrogen bonding with substrate that improves collagenolytic and general catalytic efficiency that could be exploited for new collagenase-specific inhibitor drugs. PMID- 15533939 TI - Inositol hexakisphosphate kinase-2, a physiologic mediator of cell death. AB - Diphosphoinositol pentakisphosphate (InsP7) and bis-diphosphoinositol tetrakisphosphate contain pyrophosphate bonds. InsP7 is formed from inositol hexakisphosphate (InsP6) by a family of three inositol hexakisphosphate kinases (InsP6K). In this study we establish one of the InsP6Ks, InsP6K2, as a physiologic mediator of cell death. Overexpression of wild-type InsP6K2 augments the cytotoxic actions of multiple cell stressors in diverse cell lines, whereas transfection with a dominant negative InsP6K2 decreases cell death. During cell death, InsP6 kinase activity is enhanced, and intracellular InsP7 level is augmented. Deletion of InsP6K2 but not the other forms of InsP6K diminishes cell death, suggesting that InsP6K2 is the major InsP6 kinase involved in cell death. Cytotoxicity is associated with a translocation of InsP6K2 from nuclei to mitochondria, whereas the intracellular localization of the other isoforms of the enzyme does not change. The present study provides compelling evidence that endogenous InsP6K2, by generating InsP7, provides physiologic regulation of the apoptotic process. PMID- 15533940 TI - Huntingtin-interacting protein 1 (Hip1) and Hip1-related protein (Hip1R) bind the conserved sequence of clathrin light chains and thereby influence clathrin assembly in vitro and actin distribution in vivo. AB - Clathrin heavy and light chains form triskelia, which assemble into polyhedral coats of membrane vesicles that mediate transport for endocytosis and organelle biogenesis. Light chain subunits regulate clathrin assembly in vitro by suppressing spontaneous self-assembly of the heavy chains. The residues that play this regulatory role are at the N terminus of a conserved 22-amino acid sequence that is shared by all vertebrate light chains. Here we show that these regulatory residues and others in the conserved sequence mediate light chain interaction with Hip1 and Hip1R. These related proteins were previously found to be enriched in clathrin-coated vesicles and to promote clathrin assembly in vitro. We demonstrate Hip1R binding preference for light chains associated with clathrin heavy chain and show that Hip1R stimulation of clathrin assembly in vitro is blocked by mutations in the conserved sequence of light chains that abolish interaction with Hip1 and Hip1R. In vivo overexpression of a fragment of clathrin light chain comprising the Hip1R-binding region affected cellular actin distribution. Together these results suggest that the roles of Hip1 and Hip1R in affecting clathrin assembly and actin distribution are mediated by their interaction with the conserved sequence of clathrin light chains. PMID- 15533941 TI - Huntingtin interacting protein 1 (HIP1) regulates clathrin assembly through direct binding to the regulatory region of the clathrin light chain. AB - Huntingtin interacting protein 1 (HIP1) is a component of clathrin coats. We previously demonstrated that HIP1 promotes clathrin assembly through its central helical domain, which binds directly to clathrin light chains (CLCs). To better understand the relationship between CLC binding and clathrin assembly we sought to dissect this interaction. Using C-terminal deletion constructs of the HIP1 helical domain, we identified a region between residues 450 and 456 that is required for CLC binding. Within this region, point mutations showed the importance of residues Leu-451, Leu-452, and Arg-453. Mutants that fail to bind CLC are unable to promote clathrin assembly in vitro but still mediate HIP1 homodimerization and heterodimerization with the family member HIP12/HIP1R. Moreover, HIP1 binding to CLC is necessary for HIP1 targeting to clathrin-coated pits and clathrin-coated vesicles. Interestingly, HIP1 binds to a highly conserved region of CLC previously demonstrated to regulate clathrin assembly. These results suggest a role for HIP1/CLC interactions in the regulation of clathrin assembly. PMID- 15533942 TI - Characterization of Arabidopsis fluoroacetate-resistant mutants reveals the principal mechanism of acetate activation for entry into the glyoxylate cycle. AB - The toxic acetate analogue monofluoroacetic acid was employed to isolate Arabidopsis tDNA-tagged plants deficient in their ability to utilize or sense acetate. Several tDNA-tagged lines were isolated, including two that were determined to be allelic to an EMS-mutagenized line denoted acn1 for ac non utilizing. Following conventions, the tDNA-tagged mutants were designated acn1-2 and acn1-3. Both mutants displayed identical behavior to acn1-1 on a variety of fluorinated and nonfluorinated organic acids, indicating that resistance was specific to fluoroacetate. Thermal asymmetric interlaced PCR identified the sites of tDNA insertion in both mutants to be within different exons in a gene, which encoded a protein containing an AMP-binding motif. Reverse transcription-PCR confirmed that the gene was not expressed in the mutants, and quantitative reverse transcription-PCR showed that the gene is expressed in imbibed seeds and increases in amount during establishment. The wild type AMP-binding protein cDNA was cloned and expressed in Escherichia coli, and the expressed protein was purified by nickel chelate chromatography. The enzyme was identified as an acyl CoA synthetase that was more active with acetate than butyrate and was not active with fatty acids longer than C-4. The enzyme was localized to peroxisomes by enzymatic analysis of organellar fractions isolated by sucrose density gradient centrifugation. Labeling studies with [(14)C]acetate showed that acn1 seedlings, like those of the isocitrate lyase mutant icl-1 (isocitrate lyase), are compromised in carbohydrate synthesis, indicating that this enzyme is responsible for activating exogenous acetate to the coenzyme A form for entry into the glyoxylate cycle. PMID- 15533943 TI - Macropinocytosis is the endocytic pathway that mediates macrophage foam cell formation with native low density lipoprotein. AB - Previously, we reported that fluid-phase endocytosis of native LDL by PMA activated human monocytederived macrophages converted these macrophages into cholesterol-enriched foam cells (Kruth, H. S., Huang, W., Ishii, I., and Zhang, W. Y. (2002) J. Biol. Chem. 277, 34573-34580). Uptake of fluid by cells can occur either by micropinocytosis within vesicles (<0.1 microm diameter) or by macropinocytosis within vacuoles ( approximately 0.5-5.0 microm) named macropinosomes. The current investigation has identified macropinocytosis as the pathway for fluid-phase LDL endocytosis and determined signaling and cytoskeletal components involved in this LDL endocytosis. The phosphatidylinositol 3-kinase inhibitor, LY294002, which inhibits macropinocytosis but does not inhibit micropinocytosis, completely blocked PMA-activated macrophage uptake of fluid and LDL. Also, nystatin and filipin, inhibitors of micropinocytosis from lipid-raft plasma membrane domains, both failed to inhibit PMA-stimulated macrophage cholesterol accumulation. Time-lapse video phase-contrast microscopy and time lapse digital confocal-fluorescence microscopy with fluorescent DiI-LDL showed that PMA-activated macrophages took up LDL in the fluid phase by macropinocytosis. Macropinocytosis of LDL depended on Rho GTPase signaling, actin, and microtubules. Bafilomycin A1, the vacuolar H+-ATPase inhibitor, inhibited degradation of LDL and caused accumulation of undegraded LDL within macropinosomes and multivesicular body endosomes. LDL in multivesicular body endosomes was concentrated >40-fold over its concentration in the culture medium consistent with macropinosome shrinkage by maturation into multivesicular body endosomes. Macropinocytosis of LDL taken up in the fluid phase without receptor mediated binding of LDL is a novel endocytic pathway that generates macrophage foam cells. Macropinocytosis in macrophages and possibly other vascular cells is a new pathway to target for modulating foam cell formation in atherosclerosis. PMID- 15533944 TI - Interaction of checkpoint proteins Hus1/Rad1/Rad9 with DNA base excision repair enzyme MutY homolog in fission yeast, Schizosaccharomyces pombe. AB - The DNA glycosylase MutY homolog (MYH) is responsible for removing adenines misincorporated opposite DNA strands containing guanine or 7,8-dihydro-8 oxoguanine by base excision repair thereby preventing G:C to T:A mutations. MYH has been shown to interact with the proliferating cell nuclear antigen (PCNA) in both human and fission yeast Schizosaccharomyces pombe systems. Here we show that S. pombe (Sp) MYH physically interacts with all subunits of the PCNA-like checkpoint protein heterotrimer, SpRad9/SpRad1/SpHus1, in yeast extracts and when the individual subunits are expressed in bacteria. The SpHus1 and SpPCNA binding sites are located in discrete regions of SpMYH. Immunoprecipitation assays reveal that the interaction between SpHus1 and SpMYH increases dramatically after hydrogen peroxide treatment, and this increase in the SpHus1-SpMYH interaction correlates with the presence of SpHus1 phosphorylation. In contrast, the interaction between SpPCNA and SpMYH after hydrogen peroxide treatment remains nearly unchanged. SpMYH associates with SpHus1 in a complex of approximately 450 kDa, the reported native molecular mass of the SpRad9/SpRad1/SpHus1-MYC complex. A larger portion of SpMYH shifts to the 150-500-kDa regions after hydrogen peroxide treatment in comparison with untreated extracts. SpHus1 phosphorylation is substantially reduced in SpMYH Delta cells after hydrogen peroxide treatment. These data suggest that MYH may act as an adaptor to recruit checkpoint proteins to the DNA lesions. PMID- 15533945 TI - Regulation of upstream binding factor 1 activity by insulin-like growth factor I receptor signaling. AB - The upstream binding factor 1 (UBF1) is one of the proteins in a complex that regulates the activity of RNA polymerase I, which controls the rate of ribosomal RNA (rRNA) synthesis. We have shown previously that insulin receptor substrate-1 (IRS-1) can translocate to the nuclei and nucleoli of cells and bind UBF1. We report here that activation of the type I insulin-like growth factor receptor (IGF-IR) by IGF-I increases transcription from the ribosomal DNA (rDNA) promoter in both myeloid cells and mouse fibroblasts. The increased activity of the rDNA promoter is accompanied by increased phosphorylation of UBF1, a requirement for UBF1 activation. Phosphorylation occurs on a number of UBF1 peptides, most prominently on the highly acidic, serine-rich C terminus. In myeloid cells (but not in mouse embryo fibroblasts) IRS-1 signaling stabilizes the levels of UBF1 protein. These findings demonstrate that IGF-IR signaling can increase the activity of UBF1 and transcription from the rDNA promoter, providing one explanation for the reported effects of the IGF/IRS-1 axis on cell and body size in animals and cells in culture. PMID- 15533946 TI - Kinetics of nucleotide incorporation opposite DNA bulky guanine N2 adducts by processive bacteriophage T7 DNA polymerase (exonuclease-) and HIV-1 reverse transcriptase. AB - Six oligonucleotides with carcinogen derivatives bound at the N2 atom of deoxyguanosine were prepared, including adducts derived from butadiene, acrolein, crotonaldehyde, and styrene, and examined for effects on the replicative enzymes bacteriophage DNA polymerase T7- (T7-) and HIV-1 reverse transcriptase for comparison with previous work on smaller DNA adducts. All of these adducts strongly blocked dCTP incorporation opposite the adducts. dATP was preferentially incorporated opposite the acrolein and crotonaldehyde adducts, and dTTP incorporation was preferred at the butadiene- and styrene-derived adducts. Steady state kinetic analysis indicated that the reduced catalytic efficiency with adducted DNA involved both an increased Km and attenuated kcat. Fluorescence estimates of Kd and pre-steady-state kinetic measurements of koff showed no significantly decreased affinity of T7- with the adducted oligonucleotides or the dNTP. Pre-steady-state kinetics showed no burst phase kinetics for dNTP incorporation with any of the modified oligonucleotides. These results indicate that phosphodiester bond formation or a conformational change of the enzyme.DNA complex is rate-limiting instead of the step involving release of the oligonucleotide. Thio elemental effects for dNTP incorporation were generally relatively small but variable, indicating that the presence of adducts may sometimes make phosphodiester bond formation rate-limiting but not always. PMID- 15533947 TI - DNA methyltransferase gene dDnmt2 and longevity of Drosophila. AB - The DNA methylation program of the fruit fly Drosophila melanogaster is carried out by the single DNA methyltransferase gene dDnmt2, the function of which is unknown before. We present evidence that intactness of the gene is required for maintenance of the normal life span of the fruit flies. In contrast, overexpression of dDnmt2 could extend Drosophila life span. The study links the Drosophila DNA methylation program with the small heatshock proteins and longevity/aging and has interesting implication on the eukaryotic DNA methylation programs in general. PMID- 15533948 TI - Evaluation of shoulder integrity in space: first report of musculoskeletal US on the International Space Station. AB - Investigative procedures were approved by Henry Ford Human Investigation Committee and NASA Johnson Space Center Committee for Protection of Human Subjects. Informed consent was obtained. Authors evaluated ability of nonphysician crewmember to obtain diagnostic-quality musculoskeletal ultrasonographic (US) data of the shoulder by following a just-in-time training algorithm and using real-time remote guidance aboard the International Space Station (ISS). ISS Expedition-9 crewmembers attended a 2.5-hour didactic and hands-on US training session 4 months before launch. Aboard the ISS, they completed a 1-hour computer-based Onboard Proficiency Enhancement program 7 days before examination. Crewmembers did not receive specific training in shoulder anatomy or shoulder US techniques. Evaluation of astronaut shoulder integrity was done by using a Human Research Facility US system. Crew used special positioning techniques for subject and operator to facilitate US in microgravity environment. Common anatomic reference points aided initial probe placement. Real-time US video of shoulder was transmitted to remote experienced sonologists in Telescience Center at Johnson Space Center. Probe manipulation and equipment adjustments were guided with verbal commands from remote sonologists to astronaut operators to complete rotator cuff evaluation. Comprehensive US of crewmember's shoulder included transverse and longitudinal images of biceps and supraspinatus tendons and articular cartilage surface. Total examination time required to guide astronaut operator to acquire necessary images was approximately 15 minutes. Multiple arm and probe positions were used to acquire dynamic video images that were of excellent quality to allow evaluation of shoulder integrity. Postsession download and analysis of high-fidelity US images collected onboard demonstrated additional anatomic detail that could be used to exclude subtle injury. Musculoskeletal US can be performed in space by minimally trained operators by using remote guidance. This technique can be used to evaluate shoulder integrity in symptomatic crewmembers after strenuous extravehicular activities or to monitor microgravity-associated changes in musculoskeletal anatomy. Just-in-time training, combined with remote experienced physician guidance, may provide a useful approach to complex medical tasks performed by nonexperienced personnel in a variety of remote settings, including current and future space programs. PMID- 15533949 TI - Reduced cerebrospinal fluid production and intracranial pressure in mice lacking choroid plexus water channel Aquaporin-1. AB - Aquaporin-1 (AQP1) is a water channel expressed strongly at the ventricular facing surface of choroid plexus epithelium. We developed novel methods to compare water permeability in isolated choroid plexus of wild-type vs. AQP1 null mice, as well as intracranial pressure (ICP) and cerebrospinal fluid (CSF) production and absorption. Osmotically induced water transport was rapid in choroid plexus from wild-type mice and reduced by fivefold by AQP1 deletion. AQP1 deletion did not affect choroid plexus size or structure. By stereotaxic puncture of the lateral ventricle with a microneedle, ICP was 9.5 +/- 1.4 cm H2O in wild type mice and 4.2 +/- 0.4 cm H2O in AQP1 null mice. CSF production, an isosmolar fluid secretion process, was measured by a dye dilution method involving fluid collections using a second microneedle introduced into the cisterna magna. CSF production in wild-type mice was (in microl min(-1)) 0.37 +/- 0.04 (control), 0.16 +/- 0.03 (acetazolamide-treated), and 1.14 +/- 0.15 (forskolin-treated), and reduced by approximately 25% in AQP1 null mice. Pressure-dependent CSF outflow, measured from steady-state ICP at different ventricular infusion rates, was not affected by AQP1 deletion. In a model of focal brain injury, AQP1 null mice had remarkably reduced ICP and improved survival compared with wild-type mice. The reduced ICP and CSF production in AQP1 null mice provides direct functional evidence for the involvement of AQP1 in CSF dynamics, suggesting AQP1 inhibition as a novel option for therapy of elevated ICP. PMID- 15533950 TI - Establishing the background level of base oxidation in human lymphocyte DNA: results of an interlaboratory validation study. AB - Accurate measurement of low levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8 oxodGuo) in DNA is hampered by the ease with which guanine is oxidized during preparation of DNA for analysis. ESCODD, a consortium of mainly European laboratories, has attempted to minimize this artifact and to provide standard, reliable protocols for sample preparation and analysis. ESCODD has now analyzed 8 oxodGuo in the DNA of lymphocytes isolated from venous blood from healthy young male volunteers in several European countries. Two approaches were used. Analysis of 8-oxodGuo by HPLC with electrochemical detection was performed on lymphocytes from 10 groups of volunteers, in eight countries. The alternative enzymic approach was based on digestion of DNA with formamidopyrimidine DNA glycosylase (FPG) to convert 8-oxo-7,8-dihydroguanine (8-oxoGua) to apurinic sites, subsequently measured as DNA breaks using the comet assay (7 groups of volunteers, in six countries). The median concentration of 8-oxodGuo in lymphocyte DNA, calculated from the mean values of each group of subjects as determined by HPLC, was 4.24 per 10(6) guanines. The median concentration of FPG sensitive sites, measured with the comet assay, was 0.34 per 10(6) guanines. Identical samples of HeLa cells were supplied to all participants as a reference standard. The median values for 8-oxodGuo in HeLa cells were 2.78 per 10(6) guanines (by HPLC) and 0.50 per 10(6) guanines (by enzymic methods). The discrepancy between chromatographic and FPG-based approaches may reflect overestimation by HPLC (if spurious oxidation is still not completely controlled) or underestimation by the enzymic method. Meanwhile, it is clear that the true background level of base oxidation in DNA is orders of magnitude lower than has often been claimed in the past. PMID- 15533951 TI - Optimizing lung aeration in positive end-expiratory pressure. PMID- 15533952 TI - In the treatment of tuberculosis, you get what you pay for... PMID- 15533953 TI - Asthma, hypersensitivity, and coitus. PMID- 15533954 TI - I. Neurocritical care: has it come of age? PMID- 15533955 TI - II. Fast-tracking in day surgery. Is your journey to the recovery room really necessary? PMID- 15533956 TI - III. Preoperative assessment of the airway: should anaesthetists be making use of modern imaging techniques? PMID- 15533957 TI - Difficulty in advancing a tracheal tube over a fibreoptic bronchoscope: incidence, causes and solutions. PMID- 15533958 TI - Methicillin-resistant Staphylococcus aureus in the critically ill. PMID- 15533959 TI - Unrecognized malfunction in computerized patient simulators. PMID- 15533960 TI - Dexmedetomidine for resection of a large phaeochromocytoma with invasion into the inferior vena cava. PMID- 15533961 TI - Pacemakers and defibrillators: anaesthetic implications. PMID- 15533962 TI - Getting the dog moving after surgery. PMID- 15533963 TI - Clinical assessment of disease activity for canine inflammatory bowel disease. AB - Clinical indices, consisting of gastrointestinal signs and laboratory parameters, are widely used for assessing disease activity in human inflammatory bowel disease (IBD). The development of a standardized scoring index for use in dogs with IBD would be useful in the management of clinical cases, both at diagnosis and in response to medical therapy. This review provides a synopsis of current strategies used to assess IBD activity in humans and companion animals. The clinical application of a simple scoring index (e.g., canine IBD activity index [CIBDAI]) for measurement of canine IBD activity is also reviewed. PMID- 15533964 TI - Effect of a single plasma transfusion on thromboembolism in 13 dogs with primary immune-mediated hemolytic anemia. AB - Thirteen dogs with primary immune-mediated hemolytic anemia received fresh-frozen plasma within 12 hours of admission, in addition to unfractionated heparin and other therapies, such as prednisone, azathioprine, and packed red blood cell transfusion. Antithrombin activity was quantified prior to transfusion and at 30 minutes and 48 hours after transfusion. Plasma antithrombin activity did not change significantly after a single plasma transfusion. There were no deaths in the first 48 hours of treatment. Thromboembolism was identified at necropsy in six of 10 dogs that died within 12 months of admission. There was no significant difference in the incidence of thromboembolism between the current treatment group and a historical control group. PMID- 15533965 TI - Postanesthetic esophageal dysfunction in 13 dogs. AB - Thirteen dogs with postanesthetic esophageal dysfunction were identified; 10 of these animals had esophageal stricture. Regurgitation was noted in six dogs during the inciting anesthetic event. Clinical problems common to all dogs included vomiting/regurgitation and weight loss. Coughing was noted in six dogs, and aspiration pneumonia was present in four of these dogs. The associated mortality rate was 23%. The duration of symptoms ranged from 17 to 150 days, and the diagnosis was often delayed (up to 76 days from onset of clinical signs to diagnosis). Postanesthetic esophageal dysfunction was a debilitating and costly problem that developed in one dog despite current preventative treatment. PMID- 15533966 TI - Proximal radial and distal humeral osteosarcoma in 12 dogs. AB - Twelve dogs were diagnosed with osteosarcoma of the proximal radius or distal humerus from 1990 to 2002, representing 1.0% of all dogs diagnosed with appendicular osteosarcoma. The median body weight (29.8 kg) was significantly less than that of dogs with appendicular osteosarcoma at other sites. Ten dogs were treated with amputation and chemotherapy. These dogs had a metastatic rate of 60%, a median metastasis-free interval of 356 days, and a median survival time of 824 days. There were no significant differences in metastasis-free interval or survival time between dogs with osteosarcoma of the proximal radius or distal humerus and dogs with appendicular osteosarcoma at other sites. PMID- 15533967 TI - Transdermal fentanyl patches in small animals. AB - Fentanyl citrate is a potent opioid that can be delivered by the transdermal route in cats and dogs. Publications regarding transdermal fentanyl patches were obtained and systematically reviewed. Seven studies in cats and seven studies in dogs met the criteria for inclusion in this review. Dogs achieved effective plasma concentrations approximately 24 hours after patch application. Cats achieved effective plasma concentrations 7 hours after patch application. In dogs, transdermal fentanyl produced analgesia for up to 72 hours, except for the immediate 0- to 6-hour postoperative period. In cats, transdermal fentanyl produced analgesia equivalent to intermittent butorphanol administration for up to 72 hours following patch application. PMID- 15533968 TI - A simple continuous pattern using absorbable suture for perineal urethrostomy in the cat: 18 cases (2000-2002). AB - Closure with a simple continuous pattern using absorbable suture was evaluated in 18 cats that underwent perineal urethrostomy from 2000 to 2002. The perineal urethrostomy was performed in a similar manner in all the cats, and either 4-0 or 5-0 polydioxanone was used for closure. Cats were evaluated 2 weeks postoperatively, and long-term follow-up information was reviewed. In all cats, the perineal urethrostomy site was healed within 2 weeks. None of the cats developed a stricture postoperatively. Complications were not significantly different (P>0.50) from those found in a comparison group of 21 cats operated between 1997 and 2002, in which perineal urethrostomies were performed using nonabsorbable sutures that were removed postoperatively. PMID- 15533969 TI - Alapexy: an alternative technique for repair of stenotic nares in dogs. AB - Alapexy involves the creation and apposition of two small elliptical incisions, which fix the alae in an abducted position. Permanent alapexy was achieved in four of five dogs (eight of nine nostrils) that were operated. Four animals had significant improvement in their respiratory efforts because of reduced nasal obstruction. Although the alapexy procedure was somewhat more difficult to perform than simple wedge resection techniques, it may be a viable option in those animals where other techniques have failed or in animals that have dystrophic or flaccid alar cartilage. The alapexy technique may also improve the diameter of the nares compared to previously described techniques. PMID- 15533970 TI - Aplastic articular facets in a dog with intervertebral disk rupture of the 12th to 13th thoracic vertebral space. AB - A 6-year-old, female spayed Pomeranian was presented with acute hind-limb paraplegia with the presence of deep pain perception and urinary incontinence. Myelography showed a Hansen type I herniation of the12th to 13th thoracic intervertebral space (T(12-13)). Articular facets of the T(12-13) and T(13) to first lumbar vertebra (L(1)) were absent. The spinal cord was decompressed using a bilateral T(12-13) modified lateral hemilaminectomy (pediculectomy). The aplastic sites were associated with minimal instability of the vertebral column, and stabilization of the vertebral column was not required. Familiarity with this condition is important, because articular facet aplasia may cause vertebral instability and may require an adjusted surgical approach or vertebral reduction and fusion following decompression. PMID- 15533971 TI - Diagnosis of rapid eye movement sleep disorder with electroencephalography and treatment with tricyclic antidepressants in a dog. AB - A 9-month-old, female Labrador retriever mix was presented for two types of seizure-like episodes, one of which occurred only during sleep. The two types of episodes were morphologically distinct. An electroencephalogram (EEG) demonstrated that the sleep-associated episodes occurred during rapid eye movement (REM) sleep, supporting a diagnosis of a REM behavior disorder. Based on their morphology and response to antiseizure medications, the waking episodes were diagnosed as seizures. The animal was also diagnosed with an obsessive compulsive and generalized anxiety disorder. The REM behavior disorder and anxiety-related behaviors improved with tricyclic antidepressant therapy. PMID- 15533972 TI - Heart failure caused by toxoplasmosis in a fennec fox (Fennecus zerda). AB - A male fennec fox (Fennecus zerda) kit was examined for lethargy, inappetence, and weight loss. Clinical findings included respiratory distress, a gallop rhythm, and retinochoroiditis. Radiography indicated pleural effusion and cardiomegaly. Echocardiographic findings included left ventricular dilatation, low left ventricular ejection fraction, and atrioventricular valvular regurgitation. Necropsy findings were compatible with a diagnosis of congestive heart failure caused by myocarditis. Histopathology showed a disseminated infection with Toxoplasma gondii causing myocarditis, skeletal polymyositis, gastrointestinal myositis, and panuveitis. Toxoplasma-induced myocarditis should be included in the differential diagnosis of heart failure and retinochoroiditis in the fennec fox. PMID- 15533973 TI - Radiation exposure in dental radiology: a 1998 nationwide survey in Switzerland. AB - OBJECTIVES: To measure the frequencies of dental radiological examinations in Switzerland and to determine the associated collective radiation doses. METHODS: To evaluate the frequencies, a sample of 376 dental practitioners and other institutions performing dental radiology were requested to fill in questionnaires designed to measure, amongst others, frequencies of dental radiodiagnoses according to type of examination, patient age and gender, dental specialty and type of surgery. The associated collective radiation doses were determined by multiplying the relevant frequencies with dose estimates obtained from recent measurements and calculations. RESULTS: The total number of dental examinations performed in Switzerland in 1998 was 4.1 million (581 per 1000 population). Periapical, bitewing and panoramic radiographs were the most frequent types of dental examinations. The collective dose associated with dental radiology was 71 person.Sv. This amounts to an annual average effective dose to the population of 10 muSv per caput, which is in agreement with the figures reported for countries of similar healthcare level. Various features such as the age distribution of the radiographed patients, the forms of collimators used, film consumption and the use of digital imaging systems are presented. CONCLUSIONS: Several recommendations for dose reduction are made. These include the re-evaluation of the patterns and justification for prescribing some particular types of dental examinations as well as the avoidance of unnecessary irradiation by the use of rectangular collimation and high sensitivity F-speed films. PMID- 15533974 TI - Geometric alignment and chromatic calibration of serial radiographic images. AB - OBJECTIVE: To develop software for automated registration and intensity calibration of serial dental radiographs for the analysis of longitudinal changes in bone density. METHODS: Serial dental radiographs were acquired using a positioning device designed to minimize projection divergence. Each radiograph included an image of a standardized aluminium wedge. The radiographs were scanned on a flatbed scanner (AGFA Duo Scan) with a spatial resolution of 300 dpi, and pixel intensity coded in 16-bit grey scale. The intensity was calibrated using serial images of selected areas with defined thickness of the aluminium wedge. A robust B-splines multiresolution registration algorithm was implemented to overcome the acquisition misalignment. Radiographs, taken before and after periodontal therapy, were subtracted to assess bone density evolution. RESULTS: The intensity calibration decreased the maximum intensity variations between serial radiographs from 30+/-17% to 1+/-1% (mean+/-standard deviation), and improved the visual comparison between the radiographs. The registration stage allowed correcting the misalignment of the radiographs on the scanner screen and superimposing the radiography contents. The observed residual motion was about 0.02+/-0.01 mm. CONCLUSION: Very user-friendly software was developed. The manipulator needs to scan the radiographs only one time. The software performs all subsequent processing steps. PMID- 15533975 TI - Fusion of computed tomography data and optical 3D images of the dentition for streak artefact correction in the simulation of orthognathic surgery. AB - OBJECTIVE: To determine the limits of accuracy of fusion of optical three dimensional (3D) imaging and computed tomography (CT) with and without metal artefacts in an experimental setting and to show the application of this hybrid system in 3D orthognathic surgery simulation. METHODS: Ten plaster casts of dental arches were subjected to a CT scan and optical 3D surface imaging. Subsequently, the first molars in the plaster casts were supplied with metal restorations, bilaterally, and new CT scans and optical surface images were assessed. The registration of the surface data of the two imaging modalities of the study models without and with metal restorations was carried out. The mean distance between the two data sets was calculated. From a patient a CT scan of the skull as well as optical 3D images of plaster casts of the dental arches were acquired. Again the two imaging modalities were registered and virtual orthognathic surgery simulation was carried out. RESULTS: The mean distance between the corresponding data points of CT and optical 3D surface images was 0.1262+/-0.0301 mm and 0.2671+/-0.0580 mm, respectively, for the plaster casts without and with metal restorations. The differences between these data were statistically significant (P<0.0005). For the patient case a mean difference of 0.66+/-0.49 mm and 0.56+/-0.48 mm for mandible and maxilla, respectively, was calculated between CT and optical surface data. CONCLUSION: The accuracy of the fusion of 3D CT surface data and optical 3D imaging is significantly reduced by metal artefacts. However, it seems appropriate for virtual orthognathic surgery simulation, as post-operative orthodontics are performed frequently. PMID- 15533976 TI - Conventional and digital radiographic methods in the detection of simulated external root resorptions: a comparative study. AB - OBJECTIVES: The purpose of this study was to evaluate and to compare the efficacy of a conventional and a digital radiographic method in diagnosing simulated external root resorption cavities. METHODS: Human dry mandibles containing teeth were covered with bovine muscle slices in order to simulate the soft tissues. Nine teeth from each dental group were investigated. Three periapical radiographs of each tooth were taken in an orthoradial, mesioradial and distoradial aspect using conventional film (Insight Kodak F-speed; Eastman Kodak, Rochester, NY) and a digital sensor (DRS Gnatus System; Gnatus, Ribeirao Preto, Brasil). The teeth were extracted using a forceps and had 0.7 mm and 1.0 mm deep cavities prepared on their vestibular, mesial and distal surfaces at the cervical, middle and apical thirds. Following preparation, each tooth was replaced in its alveolus and new radiographs were taken. Three dental professionals, an endodontist, a radiologist and a general practioner, evaluated the images. RESULTS: A larger number of cavities (P<0.05) were detected by the digital method when compared with the conventional method, for all depths of lesions. CONCLUSIONS: The results of this study suggest that the digital radiographic method is more sensitive than conventional radiography to detect simulated external root resorption cavities. PMID- 15533977 TI - Accuracy of diagnosing occlusal caries using enhanced digital images. AB - OBJECTIVES: To compare the accuracy of detecting occlusal caries lesions on original images vs images digitally enhanced for sharpness, zoom and pseudocolour. METHODS: Four hundred images of 100 extracted third molars were examined for the presence or absence of occlusal caries by four observers, using a 5-point confidence scale. The observers had no prior knowledge of the distribution of the caries. The definitive diagnosis of caries was based on a histological assessment after sectioning the teeth. The diagnostic accuracy for each imaging mode was expressed by the area of the receiver operating characteristic (ROC) curve. Differences between the areas under the ROC curves were assessed using an analysis of variance. RESULTS: The original image had a mean ROC curve of 0.75 (95% confidence interval (CI) 0.65-0.85). The ROC curves for sharpness, zoom and pseudocolour were 0.73 (95% CI 0.63-0.83), 0.76 (95% CI 0.61-0.81) and 0.70 (95% CI 0.60-0.80), respectively. These differences were not statistically significant (P=0.96). The Cohen's kappa indicated good agreement among observers. CONCLUSIONS: In this in vitro study, the three enhanced images used did not affect the detection of occlusal caries. PMID- 15533978 TI - Quantitative analysis of MRI signal intensity as a tool for evaluating tooth pulp vitality. AB - OBJECTIVES: To assess whether it is possible to measure tooth vitality using magnetic resonance imaging (MRI). METHODS: Signal intensity measurements were conducted using T(1) and T(2) sequences at the region of interest in 211 teeth (35 patients). RESULTS: Clinical findings showed that 17.3% of the teeth were avital, whereas 82.7% were found to be vital. Neither the T(2) sequence nor the non-contrast-enhanced T(1) sequence showed significant differences between vital and avital teeth. However, the contrast-enhanced sequence and, in particular, a comparison of signal intensities between the non-contrast-enhanced T(1) sequence and the contrast-enhanced sequence showed a significant difference between vital and avital teeth. CONCLUSION: Contrast-enhanced MRI enables us to draw conclusions on pulpal perfusion in vivo. PMID- 15533979 TI - MRI characteristics of anterior disc displacement with and without reduction. AB - OBJECTIVES: The aim of this study was to investigate the differences between magnetic resonance imaging (MRI) characteristics of anterior disc displacement with reduction (ADDR) and without reduction (ADDWR). METHODS: A clinician and a radiologist, blinded to the results of the clinical examination, independently evaluated the MRI scans of 100 subjects with symptoms of temporomandibular joint (TMJ) disorders. The final sample included 88 subjects in whom there was consensus of disc displacement both between observers and between MRI and clinical examination. There were 130 joints with ADDR and 45 joints with ADDWR in the study. The MRI characteristics such as position, signal intensity and morphology of the disc, degenerative changes, effusion, scar tissue, osteonecrosis and condylar hypermobility were evaluated in the cases of ADDR and ADDWR. The chi(2) test was used to determine the differences between ADDR and ADDWR for these MRI characteristics. RESULTS: There were no significant differences between ADDR and ADDWR for effusion and degenerative changes. There were significant differences between ADDR and ADDWR for sideways displacement, disc deformation, signal intensity changes, scar tissue, osteonecrosis and condylar hypermobility. CONCLUSIONS: Degenerative changes and effusion did not appear to be markers of either ADDR or ADDWR. However, the severity of these abnormalities may be correlated with the type of internal derangement. The prevalence of sideways displacement, disc deformation, signal intensity changes, scar tissue, and osteonecrosis was greater in ADDWR than ADDR. These conditions may be considered to be indicators of more advanced and complicated stages of internal derangement. Because the percentage of subluxation was greater in ADDR, localized joint laxity and internal derangement may be correlated. PMID- 15533980 TI - The effect of quality of temporomandibular joint MR images on interrater agreement. AB - OBJECTIVES: Effects of calibration on interrater agreement in evaluating magnetic resonance (MR) images of the temporomandibular joint (TMJ) have already been examined. The objectives of the present study were to assess to what extent the quality of MR images of the TMJ influences interrater agreement and to evaluate interrater agreement with respect to image quality assessment. METHODS: Two non calibrated medical radiologists and two general dentists evaluated sagittal images of 100 TMJs for both a rating of the image quality and the performance of five diagnostic tasks. The agreement between these raters with respect to the diagnoses was evaluated. Additionally, two additional raters, calibrated during a 5 h training including the evaluation of 70 MR images, also evaluated the diagnostic aspects and the image quality, on the basis of objective criteria. The agreement between the subjective diagnoses of the non-calibrated raters and the objective diagnoses of the calibrated raters was evaluated. Afterwards, the subjective and the objective quality assessments were compared using kappa statistics. RESULTS: When good quality images were evaluated, higher kappa values were obtained for all diagnostic categories by the non-calibrated raters (mean Deltak for making diagnoses >0.1). This finding was confirmed by the value obtained for the agreement between the non-calibrated and the calibrated raters. The non-calibrated raters were in good agreement (k=0.67, standard error +/-0.09) with the calibrated raters for assessment of image quality. CONCLUSION: The present study shows that it is possible even without calibration to obtain a better interrater agreement when higher quality MR images of the TMJ are evaluated. PMID- 15533981 TI - Schwannoma in the sublingual space. AB - In this report, we present CT and MRI findings of a case of a schwannoma that developed in the floor of the oral cavity. A 49-year-old woman visited our hospital with a painless swelling in the oral floor. CT and MRI revealed a well circumscribed oval mass in the sublingual space, which showed cystic degeneration in most of the lesion. In addition, a thickened wall that strongly enhanced after injection of contrast medium and formation of fluid level were observed in the mass. The mass was removed and was histopathologically diagnosed as schwannoma. Only a few cases of schwannoma in the oral floor have been reported. However, when the characteristic findings are observed on CT and MRI, schwannoma should be added to the differential diagnosis. PMID- 15533982 TI - Craniometaphyseal dysplasia associated with obstructive sleep apnoea syndrome. AB - Craniometaphyseal dysplasia (CMD) is a genetic craniotubular bone disorder characterized by early progressive hyperostosis and sclerosis of the craniofacial bones, and abnormal modelling of the metaphyses of the tubular bones. We present the case of a patient with a confirmed history of the autosomal dominant form of CMD, associated with symptoms of obstructive sleep apnoea syndrome. Examination and imaging studies revealed several unusual features in addition to the common findings of CMD such as: bimaxillary retrusion with hyperostosis of the mental area, severe notching of the external occipital protuberance, huge occipital horn, decreased angle of the mandible with notching of the body and thickening of the areas of muscle attachment, and macrodontia. The literature and differential diagnoses are reviewed. PMID- 15533983 TI - Atypical hypercementosis versus cementoblastoma. AB - Hypercementosis is a non-neoplastic condition in which excessive cementum is deposited in continuation with the normal radicular cementum. Apart from the idiopathic nature of hypercementosis, this condition is associated with several local and systemic factors. Cementoblastoma is an odontogenic tumour characterized by the proliferation of functional cementoblasts that form a large mass of cementum or cementum-like tissue on the tooth root. Although hypercementosis and cementoblastoma are typical conditions with distinct clinical evolution, atypical cases may challenge their diagnosis. Because cementoblastoma is a neoplasm with unlimited growth potential, the usual treatment is complete surgical resection, whilst conservative treatment is recommended for hypercementosis. An atypical case of hypercementosis with similarities to cementoblastoma is reported and its differential diagnosis is discussed. PMID- 15533984 TI - Single air contrast arthrography for temporomandibular joint disorder using limited cone beam computed tomography for dental use. AB - MRI has the problem of artefacts caused by metal or body motion and is also incompatible for patients with claustrophobia. Arthrography is invasive and involves the risk of perforation or allergy to a contrast medium. This report discusses a patient with temporomandibular joint (TMJ) disorder who required soft tissue imaging of the TMJ. As she had claustrophobia and a reaction to iodine, air contrast arthrography and pumping manipulation therapy using limited cone beam computed tomography for dental use (3DX) was performed. We conclude that the 3DX examination method used in the study is practical as a diagnostic procedure and thus recommend this method to be used for patients with TMJ disorder in the presence of iodine contraindication. PMID- 15533985 TI - Bilateral bifid mandibular condyle. AB - Bilateral bifid mandibular condyles are rare and may appear as a congenital or developmental anomaly. A case of bilateral bifid mandibular condyles is reported. The patient had no history of trauma and no link was apparent with respect to the patient's medical history. In this case, the condition was an incidental panoramic radiographic finding. Magnetic resonance imaging findings revealed bilateral anterior disc displacement without reduction. The radiographic appearance of this anomaly and the literature on bilateral bifid condyles are reviewed. PMID- 15533986 TI - Bifid mandibular condyle: archaeological case report of a rare anomaly. AB - In this paper, an archaeological case of unilateral bifid mandibular condyle is presented. This uncommon anomaly is characterized by a division of the mandibular condylar head. In this case, the left condyle was divided into two articulating surfaces oriented mediolaterally; two articular facets on the anterior wall of the glenoid fossa for the double condyle were observed. The morphological and radiological analysis do not show any evidence of injuries or degenerative pathology. Taking into account the two main causes of bifid condyle suggested in the literature (traumatic and developmental), an embryopathy by teratogenic agents is proposed as a possible aetiology of the bifid condyle reported here. PMID- 15533987 TI - Negative regulation of TRPC3 channels by protein kinase C-mediated phosphorylation of serine 712. AB - TRPC3 is a nonselective cation channel member of the "canonical" transient receptor potential (TRPC) family whose members are activated by phospholipase C coupled receptors. TRPC3 can be activated by the diacylglycerol analog 1-oleoyl-2 acetyl-sn-glycerol (OAG) in a protein kinase C-independent manner. On the other hand, phorbol 12-myristate 13-acetate (PMA) inhibits OAG-mediated TRPC3 channel activation, suggesting that phosphorylation of TRPC3 by protein kinase C is a mechanism of receptor-mediated negative feedback. Here, we show PMA-induced phosphorylation of TRPC3 channels in vivo. We demonstrate by site-directed mutagenesis that a single site containing Ser(712) and conserved among all members of the TRPC family is essential for protein kinase C-mediated negative regulation of TRPC3. In human embryonic kidney 293 cells expressing a TRPC3 mutant in which Ser(712) was replaced by alanine (S712A), PMA failed to block channel activation, whereas wild-type TRPC3 activity was completely inhibited. Phosphorylation of the S712A TRPC3 mutant was not stimulated in response to PMA treatment. Furthermore, S712A TRPC3 mutant-mediated Ca(2+) entry after methacholine activation was significantly greater than that of wild-type TRPC3. These findings demonstrate a dual role for phospholipase C-generated diacylglycerol, which serves as a signal for TRPC3 activation as well as a signal for negative feedback via protein kinase C-mediated phosphorylation. PMID- 15533988 TI - Neuropsychosocial features of very mild Alzheimer's disease (CDR 0.5) and progression to dementia in a community: the Tajiri project. AB - The borderline condition between normal aging and dementia should be detected to predict further deterioration. The authors cross-sectionally analyzed neuropsychological data, memory complaints, and social activities for community dwelling older adults. The rate of decline from Clinical Dementia Rating (CDR) 0.5 to dementia during a 3-year interval was also analyzed. Short-term memory rather than long-term memory was found to be sensitive in distinguishing those with CDR 0 from those with CDR 0.5. Relatives' observations of memory decline rather than subjective memory complaints were significantly different. Participants with CDR 0.5 reported fewer problems with social activities than did their relatives. Ten of the 29 CDR 0.5 participants (34.5%) showed cognitive decline, the decliners showing lower scores on short-term memory and orientation at the baseline condition. The neuropsychological data showed CDR 0.5 to be similar to very mild Alzheimer's disease. It would be better if subjective complaints were excluded from the criteria of the borderline condition. PMID- 15533989 TI - The clock drawing test as a measure of executive dysfunction in elderly depressed patients. AB - The aims of this research were to determine whether performance on the Clock Drawing Test (CDT) could accurately distinguish between older patients with depression and older patients with depression and previously undocumented executive dysfunction and to determine if there was a correlation between CDT and depression severity. The authors studied 52 patients consecutively admitted to a geriatric psychiatry inpatient unit of a university hospital who met DSM-IV criteria for major depression or depression not otherwise specified but had no concurrent diagnosis of dementia. All the subjects completed the Mini-Mental State Examination (MMSE), Mattis Dementia Rating Scale (DRS), and the CDT, as well as the Geriatric Depression Scale (GDS). The patients were divided into 2 subgroups based on the DRS score: <129 (cognitive impairment) versus = 129. Results indicated that the depressed patients with a score of DRS <129 had significantly lower CDT scores than did patients with DRS = 129 and normal comparison subjects (P< .01). The results support the hypothesis that CDT score is lower in elderly depressed patients with executive dysfunction versus nondepressed seniors as well as depressed patients without executive dysfunction. PMID- 15533990 TI - Executive deficits in elderly patients with major unipolar depression. AB - Several studies have evaluated executive function in depressed patients, and the results vary from significant impairment relative to controls to virtually intact performances. To better comprehend executive impairment in elderly patients with major unipolar depression, the performance of 21 elderly depressed patients was compared with that of 19 elderly normal controls on executive tasks. The relationships between memory deficits and depression severity and between memory deficits and executive dysfunction were also examined. Depressed patients' performance was significantly worse than that of controls on almost all executive tasks. Their score for logical memory was significantly correlated with that for several executive tasks. Executive performance was also correlated with depression severity. Unipolar depressed patients present executive deficits. Memory failure in these patients may reflect impairment in retrieval processes, which in turn depends on executive function. Executive deficits are associated with depression severity. These results may be useful in the differential diagnosis between depression and early Alzheimer's disease. PMID- 15533991 TI - Methodology and preliminary results from the neurocognitive outcomes of depression in the elderly study. AB - A methodology is presented for following a cohort of older depressed patients to examine neurocognitive outcomes of depression. A total of 265 depressed individuals and 138 healthy, nondepressed controls age 60 and older who completed at least 1 year of follow-up data underwent periodic clinical evaluation by a geriatric psychiatrist. A subset of 141 patients and 137 controls had neuropsychological testing. A consensus panel of experts reviewed 63 depressed subjects with suspected cognitive impairment. Twenty-seven individuals in the depressed group were assigned diagnoses of dementia, including 11 with Alzheimer's disease, 8 with vascular dementia, and 8 with dementia of undetermined etiology. In addition, 25 individuals had other forms of cognitive impairment, and 11 were considered cognitively normal. Among elderly controls, 2 developed substantial cognitive impairment with clinical diagnoses of dementia. Among the depressed group, the incidence rates for dementia for this age are much higher than would be expected. These results are consistent with prior evidence linking depression and later dementia. Future studies are needed to examine neuroimaging and genetic, clinical, and social predictors of neurocognitive decline in depression. PMID- 15533992 TI - Screening for depression in nursing home palliative care patients. AB - The purpose of the this study was to evaluate the existing tools for assessing depression in nursing home patients referred to palliative care services. The patients referred to palliative care were assessed for depression by a geriatric fellow and a psychiatrist (gold standard). The questions asked by the fellow were derived from the existing validated screening scales and diagnostic tools. The psychiatrist's assessment had a strong agreement with the Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV; kappa = 0.400) and Geriatric Depression Scale (GDS; kappa = 0.462) and least agreement with the Cornell Scale for Depression in Dementia (CDS). However, depression in the most severe dementia patients (Mini-Mental State Examination = 0) was able to be assessed only by the CDS. Thus, although items from the DSM-IV and GDS may be used to ascertain depression in geriatric patients, to date there is no scale valid and reliable enough to effectively ascertain depression in the most severely demented patients. PMID- 15533993 TI - An open treatment trial of venlafaxine for elderly patients with dysthymic disorder. AB - Treatment response and side effects of venlafaxine were evaluated in an open label trial of elderly outpatients with dysthymic disorder (DD). Patients received flexible dose (up to 300 mg/d) venlafaxine (Effexor XR) for 12 weeks. Of 23 study patients, 18 completed the trial. Fourteen (60.9%) were responders in intent-to-treat analyses with the last observation carried forward, and 77.8% were responders in completer analyses. Nearly half the sample (47.8%) met criteria for remission. In the intent-to-treat sample, increased severity of depression at baseline was associated with superior response, and the presence of cardiovascular disease was associated with poorer response. Venlafaxine open label treatment was associated with fairly high response rates and generally good tolerability in elderly patients with DD. These results indicate that in elderly patients with DD, placebo-controlled trials of a dual reuptake inhibitor such as venlafaxine would be needed to assess its efficacy or to compare its efficacy to that of other antidepressants. PMID- 15533994 TI - Serum levels of soluble intercellular adhesion molecule-1 and soluble endothelial leukocyte adhesion molecule-1 in Alzheimer's disease. AB - Serum soluble intercellular adhesion molecule-1 (s-ICAM-1) and soluble E-selectin (s-ELAM-1) were evaluated in 25 patients with Alzheimer's disease (AD), 54 patients with noninflammatory neurological diseases (NIND), and 15 control subjects. Patients with AD had a higher s-ICAM-1 level compared with the NIND patients and the control subjects (P< .001 and P< .04, respectively). The presence of high s-ICAM-1 values may be related to immunological processes involved in pathogenetic mechanisms of AD. The not statistically significant values of (s-ELAM-1), a glycoprotein considered an exclusive marker of endothelial activation, compared with the NIND patients and healthy subjects (P< .47 and P< .17, respectively), seem to suggest the neural rather than the endothelial s-ICAM origin in patients with AD. PMID- 15533995 TI - Driving and dementia: a review of the literature. AB - The purpose of this article is to review the literature on the ability of individuals with dementia to drive an automobile. Based on a review of the literature, several factors were identified that may be useful in differentiating between people with dementia who presently remain safe drivers from those who have progressed to impaired driving. These factors include disease duration and severity, sex, patient self-assessment, family assessment, neuropsychological measures, findings on road evaluations, and driving simulator testing. The approach of the physician to driving and dementia is addressed, including in office screening, referral for on-road driving assessments, and the potential for physician reporting to state agencies. PMID- 15533996 TI - Balancing Akt with S6K: implications for both metabolic diseases and tumorigenesis. AB - Proper regulation of the phosphoinositide 3-kinase-Akt pathway is critical for the prevention of both insulin resistance and tumorigenesis. Many recent studies have characterized a negative feedback loop in which components of one downstream branch of this pathway, composed of the mammalian target of rapamycin and ribosomal S6 kinase, block further activation of the pathway through inhibition of insulin receptor substrate function. These findings form a novel basis for improved understanding of the pathophysiology of metabolic diseases (e.g., diabetes and obesity), tumor syndromes (e.g., tuberous sclerosis complex and Peutz-Jegher's syndrome), and human cancers. PMID- 15533997 TI - The two cytochrome c species, DC3 and DC4, are not required for caspase activation and apoptosis in Drosophila cells. AB - In Drosophila, activation of the apical caspase DRONC requires the apoptotic protease-activating factor homologue, DARK. However, unlike caspase activation in mammals, DRONC activation is not accompanied by the release of cytochrome c from mitochondria. Drosophila encodes two cytochrome c proteins, Cytc-p (DC4) the predominantly expressed species, and Cytc-d (DC3), which is implicated in caspase activation during spermatogenesis. Here, we report that silencing expression of either or both DC3 and DC4 had no effect on apoptosis or activation of DRONC and DRICE in Drosophila cells. We find that loss of function mutations in dc3 and dc4, do not affect caspase activation during Drosophila development and that ectopic expression of DC3 or DC4 in Drosophila cells does not induce caspase activation. In cell-free studies, recombinant DC3 or DC4 failed to activate caspases in Drosophila cell lysates, but remarkably induced caspase activation in extracts from human cells. Overall, our results argue that DARK-mediated DRONC activation occurs independently of cytochrome c. PMID- 15533998 TI - A polyalanine tract expansion in Arx forms intranuclear inclusions and results in increased cell death. AB - A growing number of human disorders have been associated with expansions of a tract of a single amino acid. Recently, polyalanine (polyA) tract expansions in the Aristaless-related homeobox (ARX) protein have been identified in a subset of patients with infantile spasms and mental retardation. How alanine expansions in ARX, or any other transcription factor, cause disease have not been determined. We generated a series of polyA expansions in Arx and expressed these in cell culture and brain slices. Transfection of these constructs results in nuclear protein aggregation, filamentous nuclear inclusions, and an increase in cell death. These inclusions are ubiquitinated and recruit Hsp70. Coexpressing Hsp70 decreases the percentage of cells with nuclear inclusions. Finally, we show that expressing mutant Arx in mouse brains results in neuronal nuclear inclusion formation. Our data suggest expansions in one of the ARX polyA tracts results in nuclear protein aggregation and an increase in cell death; likely underlying the pathogenesis of the associated infantile spasms and mental retardation. PMID- 15533999 TI - A malaria membrane skeletal protein is essential for normal morphogenesis, motility, and infectivity of sporozoites. AB - Membrane skeletons are structural elements that provide mechanical support to the plasma membrane and define cell shape. Here, we identify and characterize a putative protein component of the membrane skeleton of the malaria parasite. The protein, named PbIMC1a, is the structural orthologue of the Toxoplasma gondii inner membrane complex protein 1 (TgIMC1), a component of the membrane skeleton in tachyzoites. Using targeted gene disruption in the rodent malaria species Plasmodium berghei, we show that PbIMC1a is involved in sporozoite development, is necessary for providing normal sporozoite cell shape and mechanical stability, and is essential for sporozoite infectivity in insect and vertebrate hosts. Knockout of PbIMC1a protein expression reduces, but does not abolish, sporozoite gliding locomotion. We identify a family of proteins related to PbIMC1a in Plasmodium and other apicomplexan parasites. These results provide new functional insight in the role of membrane skeletons in apicomplexan parasite biology. PMID- 15534000 TI - Apoptotic pathways are selectively activated by granzyme A and/or granzyme B in CTL-mediated target cell lysis. AB - Purified cytolytic T lymphocyte (CTL) proteases granzyme (gzm)A and gzmB with sublytic dose of perforin (perf) initiate distinct proapoptotic pathways. Their physiological relevance in CTL-mediated target cell apoptosis is elusive. Using ex vivo virus-immune CD8(+) T cells from mice deficient in perf, gzmA and/or gzmB, and the Fas-resistant EL4.F15 tumor target cell, we show that (a) CTL from gzmA(-/-) or gzmB(-/-) mice similarly induced early proapoptotic features, such as phosphatidyl serine (PS) exposure on plasma membrane, Delta Psi(m) loss, and reactive oxygen radical generation, though with distinct kinetics; (b) CTL from gzmA(-/-) but not from gzmB(-/-) mice activate caspase 3 and 9; (c) PS exposure induced by CTL from gzmA(-/-) or gzmB(-/-) mice is prevented, respectively, by caspase inhibitors or by reactive oxygen scavengers without interfering with target cell death; and (d) all gzm-induced apoptotic features analyzed depend critically on perf. Thus, perf is the principal regulator in CTL-mediated and gzm facilitated intracellular processes. The ability of gzmA and gzmB to induce multiple independent cell death pathways may be the hosts response to circumvent evasion strategies of pathogens and tumors. PMID- 15534001 TI - The ERBB4/HER4 receptor tyrosine kinase regulates gene expression by functioning as a STAT5A nuclear chaperone. AB - In the lactating breast, ERBB4 localizes to the nuclei of secretory epithelium while regulating activities of the signal transducer and activator of transcription (STAT) 5A transcription factor essential for milk-gene expression. We have identified an intrinsic ERBB4 NLS (residues 676-684) within the ERBB4 intracellular domain (4ICD) that is essential for nuclear accumulation of 4ICD. To determine the functional significance of 4ICD nuclear translocation in a physiologically relevant system, we have demonstrated that cotransfection of ERBB4 and STAT5A in a human breast cancer cell line stimulates beta-casein promoter activity. Significantly, nuclear localization of STAT5A and subsequent stimulation of the beta-casein promoter requires nuclear translocation of 4ICD. Moreover, 4ICD and STAT5A colocalize within nuclei of heregulin beta 1 (HRG) stimulated cells and both proteins bind to the endogenous beta-casein promoter in T47D breast cancer cells. Together, our results establish a novel molecular mechanism of transmembrane receptor signal transduction involving nuclear cotranslocation of the receptor intracellular domain and associated transcription factor. Subsequent binding of the two proteins at transcription factor target promoters results in activation of gene expression. PMID- 15534002 TI - Localized Ras signaling at the leading edge regulates PI3K, cell polarity, and directional cell movement. AB - During chemotaxis, receptors and heterotrimeric G-protein subunits are distributed and activated almost uniformly along the cell membrane, whereas PI(3,4,5)P(3), the product of phosphatidylinositol 3-kinase (PI3K), accumulates locally at the leading edge. The key intermediate event that creates this strong PI(3,4,5)P(3) asymmetry remains unclear. Here, we show that Ras is rapidly and transiently activated in response to chemoattractant stimulation and regulates PI3K activity. Ras activation occurs at the leading edge of chemotaxing cells, and this local activation is independent of the F-actin cytoskeleton, whereas PI3K localization is dependent on F-actin polymerization. Inhibition of Ras results in severe defects in directional movement, indicating that Ras is an upstream component of the cell's compass. These results support a mechanism by which localized Ras activation mediates leading edge formation through activation of basal PI3K present on the plasma membrane and other Ras effectors required for chemotaxis. A feedback loop, mediated through localized F-actin polymerization, recruits cytosolic PI3K to the leading edge to amplify the signal. PMID- 15534003 TI - Live cell imaging of the assembly, disassembly, and actin cable-dependent movement of endosomes and actin patches in the budding yeast, Saccharomyces cerevisiae. AB - Using FM4-64 to label endosomes and Abp1p-GFP or Sac6p-GFP to label actin patches, we find that (1) endosomes colocalize with actin patches as they assemble at the bud cortex; (2) endosomes colocalize with actin patches as they undergo linear, retrograde movement from buds toward mother cells; and (3) actin patches interact with and disassemble at FM4-64-labeled internal compartments. We also show that retrograde flow of actin cables mediates retrograde actin patch movement. An Arp2/3 complex mutation decreases the frequency of cortical, nonlinear actin patch movements, but has no effect on the velocity of linear, retrograde actin patch movement. Rather, linear actin patch movement occurs at the same velocity and direction as the movement of actin cables. Moreover, actin patches require actin cables for retrograde movements and colocalize with actin cables as they undergo retrograde movement. Our studies support a mechanism whereby actin cables serve as "conveyor belts" for retrograde movement and delivery of actin patches/endosomes to FM4-64-labeled internal compartments. PMID- 15534004 TI - Recycling endosomes can serve as intermediates during transport from the Golgi to the plasma membrane of MDCK cells. AB - The AP-1B clathrin adaptor complex is responsible for the polarized transport of many basolateral membrane proteins in epithelial cells. Localization of AP-1B to recycling endosomes (REs) along with other components (exocyst subunits and Rab8) involved in AP-1B-dependent transport suggested that RE might be an intermediate between the Golgi and the plasma membrane. Although the involvement of endosomes in the secretory pathway has long been suspected, we now present direct evidence using four independent methods that REs play a role in basolateral transport in MDCK cells. Newly synthesized AP-1B-dependent cargo, vesicular stomatitis virus glycoprotein G (VSV-G), was found by video microscopy, immunoelectron microscopy, and cell fractionation to enter transferrin-positive REs within a few minutes after exit from the trans-Golgi network. Although transient, RE entry appears essential because enzymatic inactivation of REs blocked VSV-G delivery to the cell surface. Because an apically targeted VSV-G mutant behaved similarly, these results suggest that REs not only serve as an intermediate but also as a common site for polarized sorting on the endocytic and secretory pathways. PMID- 15534006 TI - Managing transitions and ensuring good care. PMID- 15534005 TI - Protein kinase C spatially and temporally regulates gap junctional communication during human wound repair via phosphorylation of connexin43 on serine368. AB - Phosphorylation of connexin43 (Cx43) on serine368 (S368) has been shown to decrease gap junctional communication via a reduction in unitary channel conductance. Examination of phosphoserine368 (pS368) in normal human skin tissue using a phosphorylation site-specific antibody showed relatively even distribution throughout the epidermal layers. However, 24 h after wounding, but not at 6 or 72 h, pS368 levels were dramatically increased in basal keratinocytes and essentially lost from suprabasal layers adjacent to the wound (i.e., within 200 microm of it). Scratch wounding of primary human keratinocytes caused a protein kinase C (PKC)-dependent increase in pS368 in cells adjacent to the scratch, with a time course similar to that found in the wounds. Keratinocytes at the edge of the scratch also transferred dye much less efficiently at 24 h, in a manner dependent on PKC. However, keratinocyte migration to fill the scratch required early (within <6 h) gap junctional communication. Our evidence indicates that PKC-dependent phosphorylation of Cx43 at S368 creates dynamic communication compartments that can temporally and spatially regulate wound healing. PMID- 15534007 TI - Improving psychiatric drug benefit management: IV. Experiences of a pharmacy advisory committee. PMID- 15534008 TI - Drug and psychosocial curricula for psychiatry residents for treatment of schizophrenia: part I. PMID- 15534009 TI - Using best practices to manage psychiatric medications under Medicaid. PMID- 15534010 TI - An outcomes-based approach to decisions about drug coverage policies in British Columbia. PMID- 15534012 TI - A systematic review of the effectiveness of community-based mental health outreach services for older adults. AB - OBJECTIVES: Psychiatric outreach services that provide mental health assessment and treatment to older adults in their homes or communities are widely promoted as improving access and outcomes for older adults. However, a systematic review of the efficacy of these services has not been done. This review evaluates the evidence base for the effectiveness of outreach services for older adults with mental illness in noninstitutional community settings. End points of interest include the ability of the outreach program to increase access to mental health services and improve psychiatric outcomes. METHODS: MEDLINE, CINAHL, PsycINFO, and Web-of-Science databases were searched for articles in English that were indexed through May 2004. Studies were included if they evaluated face-to-face psychiatric services provided to adults aged 65 and older with mental illness and if they were randomized controlled trials, quasi-experimental outcome studies, uncontrolled cohort studies, or comparisons of two or more interventions. Articles were excluded that evaluated interventions that were provided in institutional settings or that focused on persons with dementia or their caregivers. RESULTS: Fourteen studies matched all the inclusion criteria. Two studies (one controlled prospective study and one study that used a comparison group) found support for the use of gatekeepers-nontraditional referral sources in identifying socially isolated older adults with mental illness. Twelve studies (five randomized controlled trials, one quasi-experimental study, and six uncontrolled cohort studies) found that home and community-based treatment of psychiatric symptoms were associated with improved or maintained psychiatric status. All randomized controlled trials reported improved depressive symptoms, and one reported improved overall psychiatric symptoms. CONCLUSIONS: Limited data supported the effectiveness of outreach services in identifying isolated older adults with mental illness. A more substantial evidence base indicated that home based mental health treatment is effective in improving psychiatric symptoms. Studies are needed that apply more rigorous methods evaluating the efficacy of case identification models and subsequent treatment for older persons with a variety of psychiatric diagnoses. PMID- 15534013 TI - Prevalence, severity, and co-occurrence of chronic physical health problems of persons with serious mental illness. AB - OBJECTIVES: This study examined Medicaid claims forms to determine the prevalence, severity, and co-occurrence of physical illness within a representative sample of persons with serious mental illness (N=147). METHODS: Representativeness of health problems in the study sample was established through comparison with a larger sample of persons with serious mental illness enrolled in Medicaid within the same state. Standardized annual costs were then assigned to Medicaid claims diagnoses, and individual health problem severity was measured as the sum of estimated treatment costs for diagnosed conditions. RESULTS: Seventy-four percent of the study sample (N=109) had been given a diagnosis of at least one chronic health problem, and 50 percent (N=73) had been given a diagnosis of two or more chronic health problems. Of the 14 chronic health conditions surveyed, chronic pulmonary illness was the most prevalent (31 percent incidence) and the most comorbid. Persons with chronic pulmonary illness were second only to those with infectious diseases in average annual cost of treatment ($8,277). Also, 50 percent or more of participants in eight other diagnostic categories had chronic pulmonary illness. A regression analysis identified age, obesity, and substance use disorders as significant predictors of individual health problem severity. CONCLUSIONS: Risk adjustment for physical health is essential when setting performance standards or cost expectations for mental health treatment. Excluding persons with chronic health problems from mental health service evaluations restricts generalizability of research findings and may promote interventions that are inappropriate for many persons with serious mental illness. PMID- 15534014 TI - Smoking cessation services in U.S. methadone maintenance facilities. AB - OBJECTIVES: Most patients in drug treatment smoke cigarettes. This study established the prevalence and types of nicotine dependence services offered in methadone and other opioid treatment clinics in the United States. METHODS: A cross-sectional survey was conducted of all outpatient methadone maintenance clinics in the United States. One person in a leadership position from each clinic was surveyed. The 20-minute survey was collected by phone, fax, or mail, according to responder preference. RESULTS: Fifty-nine percent of the clinics (408 of 697 clinics) responded. The sample was very similar to all outpatient methadone maintenance clinics in the United States in size, region, and ownership. In the 30 days before the survey, respondents reported that their clinics provided the following services to at least one patient: 73 percent provided brief advice to quit, 18 percent offered individual or group smoking cessation counseling, and 12 percent prescribed nicotine replacement therapy. However, the services were provided to very few patients. Clinics with written guidelines that required them to address smoking were much more likely to provide services than those without guidelines. Private for-profit clinics were significantly less likely than public or private nonprofit clinics to treat nicotine dependence. Most respondents (77 percent) reported that their staff were interested in receiving training in nicotine dependence treatment, and more than half (56 percent) had at least one staff member ("champion") with a strong interest in treating nicotine dependence. CONCLUSIONS: A vast majority of methadone patients smoke; yet in the 30 days before the survey only one out of three facilities provided counseling to any patients and only one out of ten prescribed nicotine replacement therapy to any patients. A dual strategy of requiring clinics to provide comprehensive nicotine dependence services and training staff to provide these services may provide the incentive and support necessary for the widespread adoption of treatment for nicotine dependence in methadone facilities. PMID- 15534015 TI - Treatment of tobacco use in an inpatient psychiatric setting. AB - OBJECTIVE: Despite high rates of tobacco use among psychiatric patients, such patients are one of the least studied groups of smokers, and little is known about their access to cessation treatment. This study examined delivery of tobacco cessation services in a smoke-free inpatient psychiatric setting. METHODS: Medical records of 250 psychiatric inpatients who were admitted from 1998 to 2001 were randomly selected and systematically reviewed. RESULTS: A total of 105 patients (42 percent) were identified as current smokers; the mean+/-SD number of cigarettes that they smoked per day was 21+/-15. Smokers evidenced statistically greater agitation and irritability compared with nonsmokers. None of the smokers received a diagnosis of nicotine dependence or withdrawal, and smoking status was not included in treatment planning for any patient. Nicotine replacement therapy was prescribed for 59 smokers (56 percent); of these patients, 54 (92 percent) used it. Smokers who were not given a prescription for nicotine replacement therapy were more than twice as likely as nonsmokers and smokers who were given a prescription for this therapy to be discharged from the hospital against medical advice. Only one smoker was encouraged to quit smoking, referred for cessation treatment, or provided with nicotine replacement therapy on discharge. CONCLUSIONS: Psychiatric inpatients smoke at high rates, yet interventions to treat this deadly addiction are rare. Furthermore, nicotine withdrawal left unaddressed may compromise psychiatric care. PMID- 15534016 TI - American Association of Community Psychiatrists' principles for managing transitions in behavioral health services. AB - Continuous engagement in treatment and recovery services is one of the most important aspects of addressing acute episodes of severe behavioral health problems and the ongoing disabilities associated with them. Traditionally, fragmentation in systems of care has been common, and the transition from one provider, location, or intensity of service to another has not been prioritized in treatment planning. The authors describe a set of guidelines for maintaining continuity of care that was developed by the American Association of Community Psychiatrists. These guidelines embrace a progressive conceptualization of an integrated service system. For each element of the guidelines, a sample outcome indicator is presented that could be used to measure implementation. These guidelines can be used to help form transition plans, quality improvement initiatives, and program evaluations. PMID- 15534017 TI - One-year follow-up of a multiple-family-group intervention for Chinese families of patients with schizophrenia. AB - OBJECTIVES: This study tested the effectiveness of a mutual support multiple family-group intervention for schizophrenia in terms of improvements in patients' psychosocial functioning, use of mental health services, and rehospitalization compared with a psychoeducation intervention and standard care. METHODS: A controlled trial was conducted in a sample of 96 Chinese families who were caring for a relative with schizophrenia in Hong Kong. The families were randomly assigned to one of three groups: mutual support (N=32), psychoeducation (N=33), and standard care (N=31). The interventions were delivered at two psychiatric outpatient clinics over a six-month period. The mutual support and psychoeducation interventions consisted of 12 group sessions every two weeks, each lasting about two hours. The mutual support group was a peer-led group designed to provide information, emotional support, and coping skills for caregiving in stages. The psychoeducation group was a professional-led group designed to educate families about the biological basis of schizophrenia and treatment and to improve illness management and coping skills. The standard care group and the other two groups received routine psychiatric outpatient care during the intervention. Data analyses of multiple outcomes over one-year follow up were conducted on an intention-to-treat basis. RESULTS: Multivariate analyses of variance showed that the mutual support intervention was associated with consistently greater improvements in patients' functioning and rehospitalization and stable use of mental health services over the follow-up period compared with the other two interventions. CONCLUSIONS: The study provides evidence that mutual support groups can be an effective family intervention for Chinese persons with mental illness in terms of improving patients' functioning and hospitalization without increasing their use of mental health services. PMID- 15534018 TI - Forensic assertive community treatment: preventing incarceration of adults with severe mental illness. AB - OBJECTIVE: Persons with severe mental illness are overrepresented in jails and prisons in the United States. A national survey was conducted to identify assertive community treatment programs that have been modified to prevent arrest and incarceration of adults with severe mental illness who have been involved with the criminal justice system. METHODS: Members of the National Association of County Behavioral Health Directors (NACBHD) were surveyed to identify assertive community treatment programs serving persons with criminal justice histories and working closely with criminal justice agencies. Programs were identified that met three study criteria: all enrollees had a history of involvement with the criminal justice system, a criminal justice agency was the primary referral source, and a close partnership existed with a criminal justice agency to perform jail diversion. Senior representatives of each program were subsequently contacted, and a telephone survey was administered to gather information about the design and operation of the programs. RESULTS: A total of 291 of 314 NACBHD members (93 percent) responded to the survey. Sixteen programs that met the study criteria were identified in nine states. The primary referral sources for 13 of these programs (81 percent) were local jails. Eleven programs (69 percent) incorporated probation officers as members of their assertive community treatment teams. Eight programs (50 percent) had a supervised residential component, with five providing residentially based addiction treatment. Eleven of the 16 programs have begun operating since 1999. Only three programs have published outcome data on program effectiveness. CONCLUSIONS: Forensic assertive community treatment is an emerging model for preventing arrest and incarceration of adults with severe mental illness who have substantial histories of involvement with the criminal justice system. Further research is needed to establish the structure, function, and effectiveness of this developing model of service delivery. PMID- 15534019 TI - Clinical characteristics of community forensic mental health services. AB - OBJECTIVE: Community forensic mental health teams are a new service within the widening range of specialized community mental health services. The characteristics of these novel services are poorly defined. Two commonly described service models in the United Kingdom are the integrated model (forensic specialists working within community mental health teams) and the parallel model (forensic specialists working on a separate specialist team). The study reported here aimed to establish clear definitions of these service models. METHODS: A literature review and a focus group of ten service professionals were conducted to identify candidate characteristics of services in community forensic mental health teams. A total of 31 characteristics were identified and used to prepare the first-round questionnaire for the two rounds of a modified Delphi consultation, which is an expert opinion and consensus method, with a multidisciplinary panel of 32 mental health professionals experienced in community forensic work. RESULTS: Twenty-nine staff (91 percent) completed the two rounds of consultation. Thirteen service characteristics differentiated the integrated and parallel models. Key characteristics of parallel teams included having their own team base, separate referral meetings, a specialist management line, specialist supervision, protected funding, forensic psychology, good links with criminal justice systems, and capped caseloads. Integrated teams were distinguished by their close links with community mental health services and acceptance of more referrals from primary care. CONCLUSIONS: Integrated and parallel models of community forensic mental health teams differ on many service characteristics. Defining these characteristics will help in researching the pros and cons of each model in the treatment and risk management of mentally ill offenders in the community. PMID- 15534020 TI - Sexual and reproductive behaviors among persons with mental illness. AB - For this study, 200 women and men with a major mood disorder or schizophrenia were interviewed about their sexual and reproductive behaviors. The responses of the women and men were compared with those of persons from a national health survey who were matched for age and race. Compared with women from the national survey, women with mental illness had fewer pregnancies and live births but were more likely to have had a pregnancy that did not result in a live birth. Women with mental illness had more lifetime sexual partners. The findings suggest that clinicians should pay attention to patients' sexual and reproductive health. PMID- 15534021 TI - Trends in acute psychiatric inpatient care in Massachusetts. AB - This report presents the preliminary results of a longitudinal study of acute psychiatric hospitalization in the Commonwealth of Massachusetts for fiscal years 1994 to 2000. The study was a secondary analysis of data obtained through the Commonwealth's mandated case-mix reporting system, covering 42 acute psychiatric facilities and 119,284 patients. Results include a 58.4 percent increase in the patient population, accompanied by declines in both length of stay and readmission rates; increases in the number of diagnoses of depression and in the number of patient deaths; and shifts to an older population increasingly supported by Medicaid and Medicare, especially managed care programs. PMID- 15534022 TI - Risk factors for aggressive behavior among psychiatric inpatients. AB - The evaluation of a patient's potential for aggressive behavior is an important component of care on psychiatric units. In this study, the aggressive behavior of 111 new psychiatric inpatients in Taiwan was recorded by nursing staff using the Overt Aggression Scale (OAS) during the patients' hospitalization. Psychiatrists made diagnoses in accordance with DSM-IV criteria and collected information about psychosocial risk factors for aggression. On average, aggressive patients were hospitalized longer than nonaggressive patients. After duration of hospitalization was controlled for, it was found that patients with an earlier onset of illness were more likely to be aggressive than other patients. PMID- 15534023 TI - Life-course theory of readmission of substance abusers among homeless veterans. AB - This study examined outcomes of 310 female and 315 male homeless veterans who were admitted to a Department of Veterans Affairs inpatient program for dual diagnoses of a substance use disorder and another mental illness. Participants were surveyed to determine gender differences for types of transforming experiences and for types of abuse as predictors of readmission within two years. Predictors were selected primarily from life-course theory and were analyzed with Cox's proportional hazards model. Transforming experiences, such as enhanced ego identity and spiritual well-being, attenuated the effects of childhood abuses, combat exposure, and depression for both genders. Transforming experiences also reduced the risk of readmission that was associated with aggression for men and abuse that occurred either in the military or recently for women. PMID- 15534024 TI - Association between seclusion and restraint and patient-related violence. AB - This study assessed the effect of an intervention designed to reduce the use of seclusion and restraint on reported episodes of patient-related violence on an acute inpatient psychiatric service. Results showed a significant decrease in the total number of episodes of seclusion and restraint between the 12 months before and after the intervention. However, the number of episodes of assault on patients and staff increased significantly. Efforts to decrease seclusion and restraint may be accompanied by an increased risk of harm to psychiatric patients and staff, and intensive safety monitoring and staff training should accompany all such efforts. PMID- 15534025 TI - The effects of the September 11 World Trade Center attack on a man with a preexisting mental illness. AB - Research has demonstrated an association between direct exposure to traumatic events and the exacerbation of mental illness. This report describes the case of a man who had a preexisting mental illness with psychotic symptoms and no history of psychiatric hospitalization who was evacuated from the World Trade Center area after the terrorist attack of September 11, 2001. He did not receive any intensive, specialized treatment during the ensuing months as his mental state deteriorated, and he eventually required psychiatric hospitalization for a full blown psychotic episode. Given the continuing threat of terrorist attacks, recognition of preexisting psychiatric conditions and early, specialized interventions for those at risk are essential in providing effective treatment and preventing decompensation. PMID- 15534027 TI - Cognitive dissonance in the pages of Psychiatric Services. PMID- 15534028 TI - Computer-administered versus paper-and-pencil mental health surveys. PMID- 15534030 TI - Reducing assaults among hospitalized youths. PMID- 15534031 TI - Hallucinations with tolterodine. PMID- 15534033 TI - Cotard's syndrome and electroconvulsive therapy. PMID- 15534035 TI - Moving molecules: mRNA trafficking in Mammalian oligodendrocytes and neurons. AB - Numerous mRNA molecules are localized in regions of the dendrites of neurons, some moving along dendrites in response to synaptic activity. The proteins encoded by these RNAs have diverse functions, including participation in memory formation and long-term potentiation. Recent experiments have shown that a cytoplasmic RNA trafficking pathway described for oligodendrocytes also operates in neurons. Transported RNAs possess a cis-acting element that directs them to granules, which are transported along microtubules by the motor proteins kinesin and dynein. These RNA molecules are recruited to the cytoplasmic transport granules by cooperative interaction with a cognate trans-acting factor. mRNAs containing the 11-nucleotide A2RE11 or 21-nucleotide A2RE sequences bind heterogeneous nuclear ribonucleoproteins A2 and A3, which are abundant in the brain. Mutations in this cis-acting element that weaken its interaction with hnRNP A2 also interfere with RNA trafficking. Several dendritically localized mRNAs, including those encoding calcium-calmodulin-dependent protein kinase II alpha subunit and neurogranin, possess A2RE-like sequences, suggesting that they may be localized by interaction with these heterogeneous nuclear ribonucleoproteins. Calcium-calmodulin-dependent protein kinase II alpha subunit is of particular interest: Its RNA is transported in depolarized neurons, and the protein it encodes is essential for establishing long-term memory. Several other cis-acting sequences and trans-acting factors that participate in neuronal RNA localization have been discovered. PMID- 15534036 TI - Protease-activated receptors in neuronal development, neurodegeneration, and neuroprotection: thrombin as signaling molecule in the brain. AB - Protease-activated receptors (PARs) belong to the superfamily of seven transmembrane domain G protein-coupled receptors. Four PAR subtypes are known, PAR-1 to -4. PARs are highly homologous between the species and are expressed in a wide variety of tissues and cell types. Of particular interest is the role which these receptors play in the brain, with regard to neuroprotection or degeneration under pathological conditions. The main agonist of PARs is thrombin, a multifunctional serine protease, known to be present not only in blood plasma but also in the brain. PARs possess an irreversible activation mechanism. Binding of agonist and subsequent cleavage of the extracellular N-terminus of the receptor results in exposure of a so-called tethered ligand domain, which then binds to extracellular loop 2 of the receptor leading to receptor activation. PARs exhibit an extensive expression pattern in both the central and the peripheral nervous system. PARs participate in several mechanisms important for normal cellular functioning and during critical situations involving cellular survival and death. In the last few years, research on Alzheimer's disease and stroke has linked PARs to the pathophysiology of these neurodegenerative disorders. Actions of thrombin are concentration-dependent, and therefore, depending on cellular function and environment, serve as a double-edged sword. Thrombin can be neuroprotective during stress conditions, whereas under normal conditions high concentrations of thrombin are toxic to cells. PMID- 15534038 TI - Cognitive dysfunction in Parkinson's disease: the role of frontostriatal circuitry. AB - It has been known for many years that the classic motor symptoms of Parkinson's disease are accompanied by deficits of executive function that resemble those seen after frontal lobe damage in humans. What is less clear is how different components of frontostriatal circuitry contribute to these impairments. Recently, improved methods of clinical assessment and classification, combined with novel technical approaches, such as functional neuroimaging, have led to great advances in our understanding of the fundamental mechanisms that drive frontostriatal circuitry. As a direct result, it has been possible to redefine impairments of executive function in Parkinson's disease more precisely in terms of the specific neuropsychological, neuroanatomical, and psychopharmacological mechanisms involved. PMID- 15534037 TI - Plasticity in the olfactory system: lessons for the neurobiology of memory. AB - We are rapidly advancing toward an understanding of the molecular events underlying odor transduction, mechanisms of spatiotemporal central odor processing, and neural correlates of olfactory perception and cognition. A thread running through each of these broad components that define olfaction appears to be their dynamic nature. How odors are processed, at both the behavioral and neural level, is heavily dependent on past experience, current environmental context, and internal state. The neural plasticity that allows this dynamic processing is expressed nearly ubiquitously in the olfactory pathway, from olfactory receptor neurons to the higher-order cortex, and includes mechanisms ranging from changes in membrane excitability to changes in synaptic efficacy to neurogenesis and apoptosis. This review will describe recent findings regarding plasticity in the mammalian olfactory system that are believed to have general relevance for understanding the neurobiology of memory. PMID- 15534039 TI - Glucocorticoids: protectors of the brain during innate immune responses. AB - The innate immune response is a coordinated set of reactions involving cells of myeloid lineage and a network of signaling molecules. Such a response takes place in the CNS during trauma, stroke, spinal cord injury, and neurodegenerative diseases, suggesting that macrophages/microglia are the cells that perpetuate the progressive neuronal damage. However, there is accumulating evidence that these cells and their secreted proinflammatory molecules have more beneficial effects than detrimental consequences for the neuronal elements. Indeed, a timely controlled innate immune response may limit toxicity in swiftly eliminating foreign materials and debris that are known to interfere with recovery and regeneration. Each step of the immune cascade is under the tight control of stimulatory and inhibitory signals. Glucocorticoids (GCs) act as the critical negative feedback on all myeloid cells, including those present within the brain parenchyma. Because too little is like too much, both an inappropriate feedback of GCs on microglia and high circulating GC levels in stressed individuals have been associated with deleterious consequences for the brain. In this review, the authors discuss both sides of the story with a particular emphasis on the neuro protective role of endogenous GCs during immune challenges and the problems in determining whether GCs can be a good therapy for the treatment of neuropathological conditions. PMID- 15534040 TI - A neural circuit basis for spatial working memory. AB - The maintenance of a mental image in memory over a time scale of seconds is mediated by the persistent discharges of neurons in a distributed brain network. The representation of the spatial location of a remembered visual stimulus has been studied most extensively and provides the best-understood model of how mnemonic information is encoded in the brain. Neural correlates of spatial working memory are manifested in multiple brain areas, including the prefrontal and parietal association cortices. Spatial working memory ability is severely compromised in schizophrenia, a condition that has been linked to prefrontal cortical malfunction. Recent computational modeling work, in interplay with physiological studies of behaving monkeys, has begun to identify microcircuit properties and neural dynamics that are sufficient to generate memory-related persistent activity in a recurrent network of excitatory and inhibitory neurons during spatial working memory. This review summarizes recent results and discusses issues of current debate. It is argued that understanding collective neural dynamics in a recurrent microcircuit provides a key step in bridging the gap between network memory function and its underlying cellular mechanisms. Progress in this direction will shed fundamental insights into the neural basis of spatial working memory impairment associated with mental disorders. PMID- 15534041 TI - The synaptotagmins: calcium sensors for vesicular trafficking. AB - The synaptotagmin family of vesicle proteins is believed to mediate calcium dependent regulation of membrane trafficking. Detailed biochemical and in vivo studies of the most characterized isoform, synaptotagmin 1 (syt 1), have provided compelling evidence that it functions as a calcium sensor for fast neurotransmitter release at synapses. However, the function of the remaining isoforms is unclear, and multiple roles have been hypothesized for several of these. Recent evidence in Drosophila has given insight into the function of some of the remaining synaptotagmin family members. Of the five evolutionarily conserved isoforms in Drosophila, only two, syt 1 and syt 4, localize to most, if not all, synapses. The former is localized to presynaptic terminals, whereas the latter is predominantly postsynaptic. This suggests an intriguing possibility that syt 4 may mediate a postsynaptic vesicle trafficking pathway, providing a molecular basis for an evolutionarily conserved bidirectional vesicular trafficking communication system at synapses. PMID- 15534042 TI - 5-HT1A receptors, gene repression, and depression: guilt by association. AB - The serotonin system is implicated in major depression and suicide and is negatively regulated by somatodendritic 5-HT1A autoreceptors. Desensitization of 5-HT1A autoreceptors is implicated in the 2- to 3-week latency for antidepressant treatments. Alterations in 5-HT1A receptor levels are reported in depression and suicide, and gene knockout of the 5-HT1A receptor results in an anxiety phenotype, suggesting that abnormal transcriptional regulation of this receptor gene may underlie these disorders. The 5-HT1A receptor gene is negatively regulated in neurons by repressors including REST/NRSF, Freud-1, NUDR/Deaf-1, and Hes5. The association with major depression, suicide, and panic disorder of a new functional 5-HT1A polymorphism at C(-1019)G that selectively blocks repression of the 5-HT1A autoreceptor by NUDR further suggests a causative role for altered regulation of this receptor in predisposition to mental illness. The authors review evidence that altered transcription of the 5-HT1A receptor can affect the serotonin system and limbic and cortical areas, leading to predisposition to depression. PMID- 15534043 TI - Recruiting future neuroscientists: what asking the recruits can teach us. AB - Many different strategies are used to recruit students into scientific research careers, including neuroscience research. These strategies are rarely based on knowledge about students; instead, activities are selected based on their ease of implementation. The goal of the LEARN Project is to encourage high school students into mental health science research using the theme of learning and memory. One intervention the authors developed is five Web-based biographies introducing students to contemporary neuroscientist role models studying learning and memory. To guide the design of this intervention, the authors created a survey to determine where students obtain career information and who and what influences their career selection. In a convenience sample of 124 students, the authors found that almost all students use the Internet for information about careers, in addition to consulting family members and teachers. Students' career selections are influenced most by family members, teachers, and people already in the field. The most important factors students look for in their future career are money, fun, and a good match between current interests and future careers. The data affirm the value of outreach efforts that go beyond students to include a broader audience of parents and teachers who play a critical role in career selection. PMID- 15534045 TI - Noninvasive imaging of atherosclerosis: the biology behind the pictures. PMID- 15534046 TI - 18F-FDG PET for evaluation of the treatment response in patients with gastrointestinal tract lymphomas. AB - (18)F-FDG PET is highly sensitive and specific for evaluation of the treatment response of nodal and extranodal diseases in patients with malignant lymphomas. However, no data are available in the literature with regard to (18)F-FDG PET for evaluation of the treatment response in patients with lymphomas with gastrointestinal tract (GIT) involvement. This study was undertaken to investigate the usefulness of (18)F-FDG PET in monitoring the response to the treatment of lymphomas in this setting. METHODS: We retrospectively analyzed 19 patients with different types of lymphomas (10 diffuse large B-cell lymphomas, 4 follicular lymphomas, 3 mantle cell lymphomas, and 2 Hodgkin's disease) involving GIT. Among 19 patients, 4 had gastric involvement, 13 had small bowel involvement, and 2 had small bowel plus colon involvement by lymphomas. All patients underwent (18)F-FDG PET before and after the completion of therapy. The results of (18)F-FDG PET were compared with the results of CT and clinical outcome; the presence of relapse was determined on the basis of positive biopsy results or clinical follow-up data. RESULTS: Of the 19 posttreatment PET scans, 13 showed no pathologic (18)F-FDG uptake, whereas 6 showed persistent (18)F-FDG uptake. Among the 13 patients who had negative PET scans, only 1 patient (7.7%) relapsed, whereas all 6 patients (100%) who had persistent abnormal (18)F-FDG uptake on posttherapy PET scans relapsed. Posttreatment CT scans were negative for 10 patients but showed persistent disease in the remaining 9 patients. Among the 10 patients who had negative CT scans, 9 remained in remission and 1 (10%) relapsed. Of the 9 patients who showed persistent disease, 6 (67%) relapsed and 3 (33%) remained in remission after the mean follow-up of 20 mo. The sensitivity, specificity, positive and negative predictive values, and accuracy of posttherapy (18)F-FDG PET were 86%, 100%, 100%, 92%, and 95%, respectively. The corresponding values for CT were 67%, 75%, 75%, 90%, and 79%, respectively. Patients with positive (18)F-FDG PET results had statistically significantly lower disease-free survival (DFS) (0%) than did those with positive CT results (33%) (P = 0.04). There was no statistically significant difference in DFS between patients with negative (18)F-FDG PET results and patients with negative CT results. CONCLUSION: A positive (18)F-FDG PET scan after the completion of chemotherapy in patients with lymphomas with GIT involvement is a strong predictor of relapse. (18)F-FDG PET has higher diagnostic accuracy than CT in the detection of residual disease after therapy. Despite the mild physiologic (18)F-FDG uptake in the GIT, (18)F FDG PET has potential value in monitoring the response to treatment in patients with GIT lymphomas, particularly when pretreatment PET results are positive. PMID- 15534047 TI - Significance of incidental 18F-FDG accumulations in the gastrointestinal tract in PET/CT: correlation with endoscopic and histopathologic results. AB - This study was undertaken to identify the clinical value of incidentally detected lesions (IDLs) in the gastrointestinal tract (GIT) with (18)F-FDG PET/CT. METHODS: The reported database of 3,281 patients who underwent partial-body (18)F FDG PET/CT scans from April 2001 to September 2003 was reviewed. Patients with incidental (18)F-FDG accumulations in the GIT that were associated with concomitant abnormal soft-tissue density or wall thickening on the native CT were evaluated. Incidental PET/CT findings were correlated with endoscopic and histopathologic results. RESULTS: According to our selection criteria, 98 (3%) of the 3,281 patients had an IDL of the GIT on (18)F-FDG PET/CT. Correlative endoscopic findings were available in 69 (70%) of 98 patients. Of these, 13 patients (19%) were harboring newly occurring cancers of the GIT in addition to preexisting aerodigestive tract tumors (n = 12) and malignant melanoma (n = 1). Twenty-nine (42%) patients were identified with precancerous lesions, such as advanced colonic adenomas (n = 27), Barrett's esophagus (n = 1), and intestinal metaplasia of the gastric mucosa (n = 1). Inflammatory and other benign GIT lesions were detected in 12 (17%) and 6 (8%) patients, respectively. In 9 (13%) patients, PET/CT was false-positive, showing normal findings in subsequent endoscopic examinations. In 20 (28%) of 69 patients, PET/CT findings had a relevant impact on the clinical management. Twenty-nine (30%) of the 98 patients were not subject to a further endoscopic examination because of the extent and nature of the primary tumor (n = 17), loss to follow-up (n = 7), death shortly after PET (n = 3), and patient unwillingness (n = 2). CONCLUSION: Although IDLs of the GIT on (18)F-FDG PET/CT scans are found only in about 3% of cases, they are associated with a substantial risk of an underlying cancerous or precancerous lesion. Early identification of these occult lesions may have a major impact on the patients' management and outcome. PMID- 15534048 TI - Metabolic rates in small brain nuclei determined by high-resolution PET. AB - Identification of small nuclei in the brain by PET has been limited by the spatial resolution of conventional scanners. The new detector technology and advanced signal analysis of a high-resolution research tomograph (HRRT) has improved 3-dimensional spatial resolution to 2.2 mm at sufficient efficiency and permitted the quantification of tracer concentrations in small volumes. METHODS: In 9 healthy volunteers, cerebral glucose metabolism was investigated after intravenous injection of 370 MBq of (18)F-FDG, and regional cerebral metabolic rates for glucose (rCMRGlc) were determined in various structures of the brain identified on coregistered MR images using stereotactic and topographic anatomic information. RESULTS: rCMRGlc values (in mumol/100 g/min) were higher in the cerebral cortex (33.5 +/- 2.98), the basal ganglia (32.6 +/- 3.04 in the nucleus caudatus and 40.2 +/- 3.50 in the putamen), the thalamus (36.6 +/- 4.72), and the cerebellum (29.8 +/- 2.20) and were lower in the cerebral white matter (12.3 +/- 1.45) than those reported previously with conventional scanners. This resulted in an increased ratio of cortical values to white-matter values. Various nuclei in the basal frontal lobe (21.4 +/- 3.19 in the basal forebrain and 32.3 +/- 2.39 in the nucleus accumbens), the temporal lobe (22.2 +/- 1.74 in the corpus amygdalae), the hippocampus (25.7 +/- 2.11), the diencephalon (23.1 +/- 3.33 in the corpus geniculatum laterale, 20.2 +/- 2.87 in the corpus geniculatum mediale, and 25.2 +/- 3.29 in the nucleus subthalamicus), and the brain stem (24.4 +/- 2.47 in the colliculus superior, 31.4 +/- 3.63 in the colliculus inferior, 31.0 +/- 3.10 in the nucleus ruber, and 22.8 +/- 2.35 in the substantia nigra) could be identified, and the metabolic rate was assessed in these structures. The effect of improved spatial resolution on quantified metabolic rates could directly be demonstrated in a few cases investigated on scanners of different generations. CONCLUSION: The improved spatial resolution of the HRRT decreased partial-volume effects in the quantification of metabolic rates in the brain and increased the accuracy of rCMRGlc values in large structures. For the first time, this scanner has permitted the determination of metabolic rates in small nuclei that are involved in various neurodegenerative disorders. PMID- 15534049 TI - Evaluation of 18F-FDG uptake and arterial wall calcifications using 18F-FDG PET/CT. AB - Glucose metabolic activity expressed as (18)F-FDG uptake may be increased in active atherosclerotic plaque. Calcium depositions are often increased in mature atherosclerotic plaque. The purpose of the present study was to assess the patterns of vascular-wall (18)F-FDG uptake and CT calcifications using combined PET/CT. METHODS: One hundred twenty-two consecutive patients over the age of 50 (47 women and 75 men; mean age, 66 +/- 9 y) undergoing whole-body (18)F-FDG PET/CT for tumor assessment were retrospectively evaluated. PET, CT, and PET/CT slices were generated for review. Abnormal vascular findings in major arteries in the chest and abdomen were categorized as PET positive (PET+), PET negative (PET ), CT positive (CT+), or CT negative (CT-). The topographic relationship between increased vascular-wall (18)F-FDG uptake on PET and the presence of calcifications on CT was assessed on PET/CT fused images, with abnormal sites further classified as PET+/CT+, PET+/CT-, or PET-/CT+. The presence of CT calcifications and increased vascular-wall (18)F-FDG uptake was correlated with age, sex, presence of cardiovascular risk factors, and cardiovascular disease. RESULTS: Abnormal findings were identified at 349 sites. CT calcifications (CT+) were observed at 320 sites (92%) of 100 patients (82%), more commonly in men (P < 0.03), in older patients (P < 0.0001), in patients with hypertension (P < 0.003) or hyperlipidemia (P < 0.04), and in smokers (P < 0.008). Increased vascular-wall (18)F-FDG uptake (PET+) was observed at 52 sites (15%) of 38 patients (31%), more commonly in men (P < 0.02), in older patients (P < 0.0001), and in patients with hypertension (P < 0.02), and was borderline in patients with cardiovascular disease (P = 0.057). PET+ and CT+ findings correlated in 12 patients, a PET+/CT- pattern was found in 18 patients, and 8 patients had increased vascular-wall (18)F-FDG uptake in sites with and without calcifications (PET+/CT+, CT-). Twenty two patients (18%) had a PET-/CT- pattern. CONCLUSION: Hybrid PET/CT can be used to identify and to correctly localize vascular-wall (18)F-FDG activity. Increased vascular-wall (18)F-FDG activity was found in 15% of sites and CT calcifications were noted in 92% of sites, with congruent findings in 7%. The clinical significance of the relationship between vascular-wall (18)F-FDG uptake and CT calcifications needs to be assessed by further prospective studies with long-term follow up. PMID- 15534050 TI - Tissue-specific effects on uptake of 99mTc-sestamibi by breast lesions: a targeted analysis of false scintigraphic diagnoses. AB - The limited spatial resolution of gamma-cameras is commonly considered the main reason for the low sensitivity of scintimammography in the detection of small carcinomas. The present study assessed whether uptake of (99m)Tc-sestambi is affected by certain tissue-specific parameters besides the size of the tumor. METHODS: Surgical specimens from 75 patients (30 benign lesions, 8 of which had shown false-positive scintigraphic findings, and 45 carcinomas, 8 of which had shown false-negative scintigraphic findings) were subjected to a distinct histopathologic/immunohistochemical reevaluation. Tissue-specific parameters (lesion size, cellular density, vascularity, signs of inflammation, proliferative activity, multidrug resistance expression, and receptor status) were visually scored and correlated with the sestamibi uptake on scintimammograms. RESULTS: A clear relationship was found between sestamibi uptake and tumor size. As previously assumed, a lesion size of less than 1 cm in diameter was found to be one reason for false-negative scintigraphic diagnoses. In addition, a low cell count, low vascularity, and absence of inflammation in carcinomas had a negative effect on uptake of the radiopharmaceutical. The decisive factor for increased tracer uptake by benign lesions was the presence of inflammatory changes. No correlation could be found between sestamibi uptake and proliferative cellular activity, multidrug resistance expression, or the receptor status of the tumor. CONCLUSION: Because all mentioned findings were statistically significant only in part, it is to be supposed that uptake of (99m)Tc-sestambi by breast lesions is determined by various tissue parameters in interaction. PMID- 15534051 TI - 18F-FDG kinetics in locally advanced breast cancer: correlation with tumor blood flow and changes in response to neoadjuvant chemotherapy. AB - The aim of this study was to characterize the biologic response of locally advanced breast cancer (LABC) to chemotherapy using (15)O-water-derived blood flow measurements and (18)F-FDG-derived glucose metabolism rate parameters. METHODS: Thirty-five LABC patients underwent PET with (15)O-water and (18)F-FDG before neoadjuvant chemotherapy and 2 mo after the initiation of treatment. Kinetic analysis for (15)O-water was performed using a single tissue compartment model to calculate blood flow; a 2-tissue compartment model was used to estimate (18)F-FDG rate parameters K(1), k(2), k(3), and the flux constant, K(i). Correlations and ratios between blood flow and (18)F-FDG rate parameters were calculated and compared with pathologic tumor response. RESULTS: Although blood flow and (18)F-FDG transport (K(1)) were correlated before chemotherapy, there was relatively poor correlation between blood flow and the phosphorylation constant (k(3)) or the overall (18)F-FDG flux (K(i)). Blood flow and (18)F-FDG flux were more closely matched after chemotherapy, with changes in k(3) accounting for the increased correlation. These findings were consistent with a decline in both the K(i)/flow and k(3)/flow ratios with therapy. The ratio of (18)F-FDG flux to transport (K(i)/K(1)) after 2 mo of chemotherapy was predictive of ultimate response. CONCLUSION: The pattern of tumor glucose metabolism in LABC, as reflected by analysis of (18)F-FDG rate parameters, changes after therapy, even in patients with modest clinical responses. This may indicate a change in tumor "metabolic phenotype" in response to treatment. A low ratio of glucose metabolism (reflected by K(i)) to glucose delivery (reflected by K(1) and blood flow) after therapy is associated with a favorable response. Further work is needed to understand the tumor biology underlying these findings. PMID- 15534052 TI - Decreased 18F-FDG uptake 1 day after initiation of chemotherapy for malignant lymphomas. AB - Several recent reports have described the judgment of chemotherapeutic effects on malignant lymphomas by use of (18)F-FDG PET as early as a few courses after the initiation of chemotherapy. However, the optimal timing of (18)F-FDG PET has yet to be clarified. Earlier (18)F-FDG PET, such as day 1 after chemotherapy, may be affected by inflammation or chemotoxicity in addition to chemotherapeutic effects, but the ways in which uptake is changed are as yet unclear. We therefore examined changes in (18)F-FDG PET results on day 1 after the initiation of chemotherapy for malignant lymphoma. METHODS: Twelve patients with non-Hodgkin's lymphoma were enrolled in this study. (18)F-FDG PET was performed before therapy to determine baseline results and then was repeated at day 1 and day 20 after the initiation of chemotherapy (just before the initiation of the second course of chemotherapy) and at the end of chemotherapy. We selected 1-9 regions of interest (ROIs) from each patient and calculated the corrected standardized uptake value (SUV(cor)) by subtracting the SUV of surrounding normal tissue for a semiquantitative analysis. From the ROIs in each patient, the representative SUV(cor) with the highest SUV(cor) at baseline was selected, and the mean representative SUV(cor)s for all 12 patients at baseline, day 1, day 20, and the end of chemotherapy were evaluated. Changes in the representative SUV(cor) were compared by use of paired t tests (2-tailed P values of <0.05 were considered statistically significant). RESULTS: All representative SUV(cor)s for each patient were lower on day 1 than at baseline, and the mean +/- SD representative SUV(cor) for all patients was significantly decreased from 10.7 +/- 7.9 at baseline to 5.8 +/- 5.8 at day 1 (P = 0.0002; paired t test). On day 20, the mean +/- SD SUV(cor) was 0.7 +/- 1.0, showing a further decrease from the value at day 1 (P = 0.01). Although the mean +/- SD SUV(cor) tended to decrease again to 0.4 +/- 0.7 by the end of chemotherapy compared with the value at day 20, no significant difference was identified (P = 0.37). CONCLUSION: (18)F-FDG uptake decreased as early as day 1 after the initiation of chemotherapy, indicating that (18)F-FDG PET for initial diagnosis or staging must be performed before the onset of chemotherapy, as scan results might already be severely compromised after the first day. PMID- 15534053 TI - 18F-FDG PET in patients with esophageal squamous cell carcinoma undergoing curative surgery: prognostic implications. AB - We investigated whether the standardized uptake value (SUV) of the primary tumor, the tumor length measured on a PET image, the number of (18)F-FDG PET-positive nodes, and the PET stage were independent prognostic predictors over other clinical variables in patients with esophageal squamous cell carcinoma who were undergoing curative surgery. METHODS: Sixty-nine patients with newly diagnosed esophageal squamous cell carcinoma who underwent preoperative (18)F-FDG PET and curative esophagectomy were included. The events for survival analysis were defined as recurrence or metastasis and cancer-related death. The disease-free and overall survival rates of each variable were estimated by the Kaplan-Meier method. The Cox proportional hazards model was used to evaluate independent prognostic variables for multivariate survival analysis. RESULTS: Using univariate survival analysis, the presence of adjuvant therapy, pathologic stage, number of CT-positive nodes (0, 1, > or =2), tumor length on PET (cutoff: 3 cm, 5 cm), number of PET-positive nodes (0, 1, 2, > or =3), and PET stage (N0 M0, N1 M0, M1) were significant prognostic predictors for disease-free survival. However, only the number of PET-positive nodes was an independent significant prognostic predictor for disease-free survival in multivariate analysis (hazard ratio = 1.87, P < 0.001). In univariate survival analysis, the sex, presence of adjuvant therapy, clinical and pathologic stages, number of CT-positive nodes, maximum SUV of the primary tumor (cutoff: 6.3, 13.7), tumor length on PET, number of PET-positive nodes, and PET stage were significant prognostic predictors for overall survival. In contrast, the clinical stage (hazard ratio = 0.53, P < 0.05), pathologic stage (hazard ratio = 3.14, P < 0.005), tumor length by PET (hazard ratio = 2.74, P = 0.01), and number of PET-positive nodes (hazard ratio = 1.71, P < 0.05) were independent significant prognostic predictors for overall survival in multivariate analysis. CONCLUSION: In addition to the pathologic stage, (18)F-FDG PET provides noninvasively independent prognostic information using the number of positive lymph nodes and the tumor length on the PET image in preoperative esophageal squamous cell carcinoma. A revised TNM classification system for esophageal carcinoma may consider tumor length and the number of positive lymph nodes as important prognostic factors. PMID- 15534054 TI - Assessment of hypoxia and perfusion in human brain tumors using PET with 18F fluoromisonidazole and 15O-H2O. AB - Hypoxia predicts poor treatment response of malignant tumors. We used PET with (18)F-fluoromisonidazole ((18)F-FMISO) and (15)O-H(2)O to measure in vivo hypoxia and perfusion in patients with brain tumors. METHODS: Eleven patients with various brain tumors were investigated. We performed dynamic (18)F-FMISO PET, including arterial blood sampling and the determination of (18)F-FMISO stability in plasma with high-performance liquid chromatography (HPLC). The (18)F-FMISO kinetics in normal brain and tumor were assessed quantitatively using standard 2- and 3-compartment models. Tumor perfusion ((15)O-H(2)O) was measured immediately before (18)F-FMISO PET in 10 of the 11 patients. RESULTS: PET images acquired 150 170 min after injection revealed increased (18)F-FMISO tumor uptake in all glioblastomas. This increased uptake was reflected by (18)F-FMISO distribution volumes >1, compared with (18)F-FMISO distribution volumes <1 in normal brain. The (18)F-FMISO uptake rate K(1) was also higher in all glioblastomas than in normal brain. In meningioma, which lacks the blood-brain barrier (BBB), a higher K(1) was observed than in glioblastoma, whereas the (18)F-FMISO distribution volume in meningioma was <1. Pixel-by-pixel image analysis generally showed a positive correlation between (18)F-FMISO tumor uptake at 0-5 min after injection and perfusion ((15)O-H(2)O) with r values between 0.42 and 0.86, whereas late (18)F-FMISO images (150-170 min after injection) were (with a single exception) independent of perfusion. Spatial comparison of (18)F-FMISO with (15)O-H(2)O PET images in glioblastomas showed hypoxia both in hypo- and hyperperfused tumor areas. HPLC analysis showed that most of the (18)F-FMISO in plasma was still intact 90 min after injection, accounting for 92%-96% of plasma radioactivity. CONCLUSION: Our data suggest that late (18)F-FMISO PET images provide a spatial description of hypoxia in brain tumors that is independent of BBB disruption and tumor perfusion. The distribution volume is an appropriate measure to quantify (18)F-FMISO uptake. The perfusion-hypoxia patterns described in glioblastoma suggest that hypoxia in these tumors may develop irrespective of the magnitude of perfusion. PMID- 15534055 TI - Educational level influences regional cerebral blood flow in patients with Alzheimer's disease. AB - We tested the hypothesis that educational level influences regional cerebral blood flow (rCBF) in Alzheimer's disease (AD) patients. METHODS: The severity of AD was measured with the Cognitive Ability Screening Instrument (age and education adjusted). rCBF was assessed using (99m)Tc-hexamethylpropyleneamine oxime brain SPECT; differences in rCBF between groups with different educational levels were determined using statistical parametric mapping (SPM). RESULTS: In matched low-education (< or =6 y; n = 29) and high-education (>6 y; n = 29) groups, SPM revealed 2 statistically significant clusters of voxels with higher rCBF in the high-education group: one in the left lateral inferior, middle, and superior temporal gyrus; another in the left medial temporal area to the left inferior frontal gyrus. CONCLUSION: We provide biologic evidence that education may lead to relatively higher rCBF in specific areas in AD patients, which may explain the effects of education on clinical manifestations of AD. PMID- 15534056 TI - Diagnosing infection in the failed joint replacement: a comparison of coincidence detection 18F-FDG and 111In-labeled leukocyte/99mTc-sulfur colloid marrow imaging. AB - The objectives of this study were to investigate (18)F-FDG imaging, using a coincidence detection system, for diagnosing prosthetic joint infection and to compare it with combined (111)In-labeled leukocyte/(99m)Tc-sulfur colloid marrow imaging in patients with failed lower extremity joint replacements. METHODS: Fifty-nine patients--with painful, failed, lower extremity joint prostheses, 40 hip and 19 knee--who underwent (18)F-FDG, labeled leukocyte, and bone marrow imaging, and had histopathologic and microbiologic confirmation of the final diagnosis, formed the basis of this investigation. (18)F-FDG images were interpreted as positive for infection using 4 different criteria: criterion 1: any periprosthetic activity, regardless of location or intensity; criterion 2: periprosthetic activity on the (18)F-FDG image, without corresponding activity on the marrow image; criterion 3: only bone-prosthesis interface activity, regardless of intensity; criterion 4: semiquantitative analysis--a lesion-to background ratio was generated, and the cutoff value yielding the highest accuracy for determining the presence of infection was determined. Labeled leukocyte/marrow images were interpreted as positive for infection when periprosthetic activity was present on the labeled leukocyte image without corresponding activity on the marrow image. RESULTS: Twenty-five (42%) prostheses, 14 hip and 11 knee, were infected. The sensitivity, specificity, and accuracy of (18)F-FDG, by criterion, were as follows: criterion 1: 100%, 9%, 47%; criterion 2: 96%, 35%, 61%; criterion 3: 52%, 44%, 47%; criterion 4: 36%, 97%, 71%. The sensitivity, specificity, and accuracy of labeled leukocyte/marrow imaging were 100%, 91%, and 95%, respectively. WBC/marrow imaging, which was more accurate than any of the (18)F-FDG criteria for all prostheses, as well as for hips and knees separately, was significantly more sensitive than criterion 3 (P < 0.001) and criterion 4 (P < 0.001) and was significantly more specific than criterion 1 (P < 0.001), criterion 2 (P < 0.001), and criterion 3 (P < 0.001). CONCLUSION: Regardless of how the images are interpreted, coincidence detection based (18)F-FDG imaging is less accurate than, and cannot replace, labeled leukocyte/marrow imaging for diagnosing infection of the failed prosthetic joint. PMID- 15534057 TI - Constancy and variability of gallbladder ejection fraction: impact on diagnosis and therapy. AB - The main objective of this study was to test the constancy and variability of gallbladder (GB) ejection fraction (EF) in long-term studies to (a) determine whether EF ever becomes normal once it is low, (b) determine how long it takes for the EF to become abnormal once it is found to be normal, (c) explore the cause of low EF, and (d) define objective parameters for biliary and nonbiliary abdominal pain. METHODS: Fifty-two patients (42 women, 10 men) who underwent quantitative cholescintigraphy twice (total studies, 104), over a mean period of 38.54 mo between studies, were chosen for retrospective analysis. They were divided into the following groups: control (n = 13; nonbiliary abdominal pain), chronic acalculous cholecystitis (CAC) (n = 27; biliary abdominal pain), chronic calculous cholecystitis (CCC) (n = 6; biliary abdominal pain), and opioid (n = 6; nonbiliary abdominal pain). The last group had received an opioid before cholecystokinin-8 (CCK-8) infusion in one study but not in the other study. A GBEF value of > or =35% was considered normal with a 3-min infusion and > or =50% as normal with a 10-min infusion of CCK-8. RESULTS: The mean GBEF value was reproducible between the 2 sequential studies in the control group (66.0% +/- 20.5% vs. 73.9% +/- 17.7%), CAC group (24.4% +/- 22.3% vs. 16.9% +/- 10.9%), and CCC group (20.8% +/- 20.9% vs. 27.5% +/- 34.5%) but not in the opioid group (14.8% +/- 14.6% vs. 56.5% +/- 31.7%). The severity of GBEF reduction in CAC increased with time: 7.2% +/- 8.1% within 12 mo, 16.1% +/- 14.9% in 13-47 mo, and 23.5% +/- 21.3% in 48-168 mo. None of the 27 patients with CAC developed a gallstone as detected by ultrasound during the study period. In 5 patients with CAC, a mean period of 52.6 +/- 28.9 mo was required for conversion from normal to a low EF. CCK-induced cystic duct spasm is the etiology for low EF in both CAC and CCC. CONCLUSION: Normal and low GBEF values are reproducible in long-term studies. Once the EF reaches a low value, it does not return to normal, and a normal value requires many years to become abnormal. CCK-induced cystic duct spasm is the cause of low GBEF in CAC and CCC, and the severity of EF reduction is similar for both. Exclusion of opioid intake immediately before the study is critical before attributing a low GBEF value to an irreversible GB motor dysfunction. PMID- 15534058 TI - Dobutamine stress 99mTc-tetrofosmin quantitative gated SPECT predicts improvement of cardiac function after carvedilol treatment in patients with dilated cardiomyopathy. AB - We evaluated whether dobutamine stress (99m)Tc-tetrofosmin quantitative gated SPECT (D-QGS) could predict improvement of cardiac function by carvedilol therapy in patients with dilated cardiomyopathy (DCM). METHODS: The study included 30 patients with idiopathic DCM and a left ventricular ejection fraction (LVEF) of <45%. D-QGS was performed in all patients to measure LVEF at rest and during dobutamine infusion (10 microg/kg/min). LVEF and left ventricular end-diastolic volume (LVEDV) were determined by echocardiography, plasma brain natriuretic peptide (BNP) was measured, and the New York Heart Association (NYHA) functional class was estimated at baseline and after 1 y of combined treatment with an angiotensin-converting enzyme (ACE) inhibitor, diuretic, and the beta-blocker carvedilol. After treatment, the echocardiographic LVEF improved by >5% in 15 patients (group A) but did not improve in the remaining 15 patients (group B). RESULTS: The baseline LVEF, LVEDV, plasma BNP, and NYHA functional class were similar in both groups. However, there was a greater increase of LVEF (Delta LVEF) with dobutamine infusion during D-QGS in group A than that in group B (12.0% +/- 5.8% vs. 2.7% +/- 4.2%, P < 0.0001). When a cutoff value of 6.6% for Delta LVEF was used to predict the improvement of LVEF by carvedilol therapy, the sensitivity was 86.7%, the specificity was 86.7%, and the accuracy was 86.7%. LVEDV, plasma BNP, and NYHA functional class all showed superior improvement in group A compared with group B. CONCLUSION: Delta LVEF measured by D-QGS was significantly larger in patients who responded to carvedilol than that in nonresponders. These findings indicate that D-QGS can be used to predict improvement of cardiac function and heart failure symptoms by carvedilol therapy in patients with idiopathic DCM. PMID- 15534059 TI - Reduced oxidative metabolic response in dysfunctional myocardium with preserved glucose metabolism but with impaired contractile reserve. AB - The recovery of function in myocardium defined as viable by (18)F-FDG PET may differ from that defined by dobutamine stress echocardiography (DSE). The aim of this study was to investigate the difference in the oxidative metabolic response between myocardial segments with preserved contractile reserve (CR) and those without CR, in segments with and without preserved glucose metabolism (GM), using (11)C-acetate PET. METHODS: Twenty patients with previous myocardial infarction (left ventricular ejection fraction, 37.1% +/- 16.5%) underwent dynamic (11)C acetate PET at rest and during dobutamine (7.5 microg/kg/min) infusion. GM was evaluated using (18)F-FDG PET and CR was evaluated using DSE. Dysfunctional segments were divided into 3 groups: group A (n = 26) with preserved CR and GM, group B (n = 15) without CR but with preserved GM, and group C (n = 41) without CR and without preserved GM. RESULTS: Resting oxidative metabolism (k mono = monoexponential clearance rate) was preserved in group A and group B (0.052 +/- 0.011/min vs. 0.051 +/- 0.012/min, P = not significant) but was reduced in group C (0.040 +/- 0.015/min) (P < 0.03 vs. group A and group B). The change in k mono, as a measure of the metabolic response to low-dose dobutamine, was significantly higher in group A (0.018 +/- 0.012) than that in group B (0.0075 +/- 0.0096, P < 0.03) and group C (0.0080 +/- 0.012, P < 0.005). CONCLUSION: Viable segments based on (18)F-FDG PET have preserved resting oxidative metabolism. However, segments without CR but with preserved GM show a reduction in the oxidative metabolic response to low-dose dobutamine infusion. The decrease in CR may be related to the reduction in the metabolic response to inotropic stimulation despite preservation of tissue viability on (18)F-FDG PET. PMID- 15534060 TI - Metabolic response after intraarterial 90Y-glass microsphere treatment for colorectal liver metastases: comparison of quantitative and visual analyses by 18F-FDG PET. AB - Our aim was to assess the feasibility of using PET for quantifying metabolic response after intraarterial (90)Y-glass microspheres for metastatic colorectal cancer to the liver by comparing visual estimates with hepatic standardized uptake values (SUVs). METHODS: Twenty-seven patients (15 men, 12 women; age, 68 +/- 12 y [+/-SD]) with metastatic colorectal cancer to the liver, and tumor progression despite polychemotherapy, were included. All patients had baseline CT or MRI, (18)F-FDG PET, hepatic angiography, and intraarterial (99m)Tc-labeled macroaggregated albumin scanning. Patients were treated with (90)Y-glass microspheres and were monitored for 3 mo using PET and serum carcinoembryonic antigen. The average absorbed dose was 139 +/- 22 Gy. All treatments were performed on a lobar basis. For each case analyzed, regions of interest were drawn along the liver edge to measure SUVs on maximum-intensity-projection (MIP) and resliced axial images. Concomitantly, the visual estimate was graded as +1, 0, -1, -2, or -3 for progression, no change, and mild, moderate, and dramatic improvement at posttreatment PET. RESULTS: Visual estimates placed 20 patients in the response category (-3 to -1) and 7 patients in the nonresponse category (0 to +1). There was a significant drop in the median SUV on the resliced axial images from 10,455 at baseline to 9,075 after treatment (P = 0.011) for the entire group. The percentage of metabolic response was significantly greater in the response group compared with that of the nonresponse group (-26% +/- 25% vs. +6% +/- 15%, P = 0.004). This correlated significantly with the respective visual estimates (r = 0.75, P < 0.0001). Furthermore, the direction of change agreed in 85% of patients using both methods. There was no significant correlation when the SUV from the simplified MIP images were used in the coronal or sagittal manner. CONCLUSION: It is feasible to quantify reduction of hepatic tumor metabolism objectively after (90)Y treatment for unresectable metastatic disease to the liver. SUVs of the entire axial slices of liver agree well with subjective visual evaluations. Quantitative PET is a useful technique in the treatment response evaluation of patients after (90)Y-glass microspheres. PMID- 15534061 TI - Molecular and metabolic imaging of atherosclerosis. AB - Atherosclerosis is a systemic disease that affects most major arteries of the body and is the most common cause of premature death in the western world. It develops slowly and often asymptomatically, so that for many patients its first manifestation is sudden cardiac death, stroke, or myocardial infarction. The current gold standard for imaging atherosclerosis is x-ray angiography. However, recent advances in understanding of the pathobiology of atherosclerosis have highlighted the inadequacies of this technique and the need for better imaging approaches. The purpose of this article is to briefly outline the biology of atherosclerosis and to review the techniques available to image it, concentrating specifically on those that detect metabolic or inflammatory changes within the atherosclerotic plaque. PMID- 15534062 TI - Improvement of algorithm for quantification of regional myocardial blood flow using 15O-water with PET. AB - (15)O-Water and dynamic PET allow noninvasive quantification of myocardial blood flow (MBF). However, complicated image analyzing procedures are required, which may limit the practicality of this approach. We have designed a new practical algorithm, which allows stable, rapid, and automated quantification of regional MBF (rMBF) using (15)O-water PET. We designed an algorithm for setting the 3 dimensional (3D) region of interest (ROI) of the whole myocardium semiautomatically. Subsequently, a uniform input function was calculated for each subject using a time-activity curve in the 3D whole myocardial ROI. The uniform input function allows the mathematically simple and robust algorithm to estimate rMBF. METHODS: Thirty-six volunteers were used in the static (15)O-CO and dynamic (15)O-water PET studies. To evaluate the reproducibility of the estimates, a repeated (15)O-water scan was obtained under resting condition. In addition, to evaluate the stability of the new algorithm in the hyperemic state, a (15)O-water scan was obtained with adenosine triphosphate. This algorithm includes a procedure for positioning a 3D ROI of the whole myocardium from 3D images and dividing it into 16 segments. Subsequently, the uniform input function was calculated using time-activity curves in the whole myocardial ROI and in the LV ROI. The uniform input function allowed this simple and robust algorithm to estimate the rMBF, perfusable tissue fraction (PTF), and spillover fraction (Va) according to a single tissue compartment model. These estimates were compared with those calculated using the original method. A simulation study was performed to compare the effects of errors in PTF or Va on the MBF using the 2 methods. RESULTS: The average operating time for positioning a whole myocardial ROI and 16 regional myocardial ROIs was <5 min. The new method yielded less deviation in rMBF (0.876 +/- 0.177 mL/min/g, coefficient of variation [CV] = 20.2%, n = 576) than those with the traditional method (0.898 +/- 0.271 mL/min/g, CV = 30.1%, n = 576) (P < 0.01). In the hyperemic state, the new method yielded less deviation in rMBF (3.890 +/- 1.250 mL/min/g, CV = 32.1%) than those with the traditional method (3.962 +/- 1.762 mL/min/g, CV = 44.4%) (P < 0.05). This method yielded significantly higher reproducibility of rMBF (r = 0.806, n = 576) than the original method (r = 0.756, n = 576) (P < 0.05). Our new method yielded a better correlation in the repeated measurement values of rMBF and less variability among the regions in the myocardium than with the original theory of the (15)O-water technique. The simulation study demonstrated fewer effects of error in the PTF or Va on the MBF value with the new method. CONCLUSION: We have developed a technique for an automated, simplified, and stable algorithm to quantify rMBF. This software is considered to be practical for clinical use in myocardial PET studies using (15)O-water with a high reproducibility and a short processing time. PMID- 15534063 TI - PET of cardiac transgene expression: comparison of 2 approaches based on herpesviral thymidine kinase reporter gene. AB - PET of reporter gene expression holds promise for noninvasive monitoring of gene therapy. Previously, 2 approaches based on the herpes simplex virus type 1 thymidine kinase gene (HSV1-tk) have been successfully applied to the heart. Wild type HSV1-tk was imaged with (124)I-labeled 2'-fluoro-2'-deoxy-5-iodo-1-beta-D arabinofuranosyl-5-iodouracil (FIAU), and a mutant HSV1-tk (HSV1-sr39tk) was imaged with (18)F-labeled 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine (FHBG). The aim of this study was to compare these 2 combinations with regard to specificity, imaging contrast, and reporter probe kinetics using dynamic PET in small and large animals. METHODS: Similar titers of adenovirus-expressing wild-type HSV1-tk (Ad(tk)), mutant HSV1-sr39tk (Ad(sr39tk)), or control genes were directly injected into the myocardium of 24 rats and 8 pigs. Two days later, dynamic PET was performed with a clinical scanner during the 120 min after injection of (124)I-FIAU (Ad(tk) animals and controls) or (18)F-FHBG (Ad(sr39tk) animals and controls). Imaging with (13)N-ammonia was performed to identify cardiac regions of interest. RESULTS: In rats, significant cardiac (124)I-FIAU accumulation occurred in images obtained early (10-30 min) after Ad(tk) injection. Because of tracer washout, however, no difference between Ad(tk)-injected animals and controls was seen in the images obtained later. For (18)F-FHBG, specific myocardial accumulation greater than background levels was detected in Ad(sr39tk) injected animals at early imaging and, in contrast to (124)I-FIAU accumulation, increased over time until the latest imaging (105-120 min). At maximum, cardiac (18)F-FHBG concentration showed a 4.15 +/- 1.65-fold increase compared with controls (105-120 min), and cardiac (124)I-FIAU concentration reached a maximal increase of 1.34 +/- 0.38-fold compared with controls (10-30 min, P = 0.0014). Global cardiac reporter probe kinetics in rats were confirmed by regional myocardial analysis in pig hearts. Transgene expression was specifically visualized by both approaches. The highest target-to-background ratio of (124)I FIAU in Ad(tk)-infected pig myocardium was 1.50 +/- 0.20, versus 2.64 +/- 0.49 for (18)F-FHBG in Ad(sr39tk)-infected areas (P = 0.01). In vivo results were confirmed by ex vivo counting and autoradiography. CONCLUSION: Both reporter gene/probe combinations were feasible for noninvasive imaging of cardiac transgene expression in different species. Specific probe kinetics suggest different myocardial handling of pyrimidine (FIAU) and acycloguanosine (FHBG) derivatives. The results favor (18)F-FHBG with mutant HSV1-sr39tk because of continuous accumulation over time and higher imaging contrast. PMID- 15534064 TI - Quantitative myocardial perfusion and coronary reserve in rats with 13N-ammonia and small animal PET: impact of anesthesia and pharmacologic stress agents. AB - The purpose of this study was to evaluate the effects of 2 anesthetic agents on myocardial perfusion and coronary reserve in rats under resting and stress conditions with small animal PET. METHODS: Twenty-four rest/stress studies were performed in 6 rats. Each animal received all 4 possible combinations of anesthetic agents (propofol, isoflurane) and pharmacologic stress agents (dobutamine, adenosine) to increase myocardial perfusion. For each stress or rest study, a 10-min dynamic acquisition was performed in list mode with 185 MBq of (13)N-NH(3). Data analysis was performed according to a 3-compartment myocardial blood flow model. Pharmacologic stimulation by either dobutamine or adenosine was performed to increase myocardial perfusion. RESULTS: The perfusion values (mean +/- SD) for the various experimental conditions were as follows: propofol/dobutamine, 7.8 +/- 2.4 mL/g/min (rest, 3.7 +/- 0.8 mL/g/min; mean +/- SD); isoflurane/dobutamine, 9.3 +/- 3.1 mL/g/min (rest, 4.3 +/- 1.0 mL/g/min); propofol/adenosine, 6.8 +/- 1.7 mL/g/min (rest, 3.2 +/- 0.4 mL/g/min); and isoflurane/adenosine, 5.2 +/- 1.3 mL/g/min (rest, 3.7 +/- 0.7 mL/g/min). All perfusion data showed a significant increase after pharmacologic stimulation relative to baseline (P < 0.05). The coronary reserve (mean +/- SD) measured by PET was slightly lower with the combination of isoflurane and adenosine (1.4 +/- 0.5) than with propofol and adenosine (2.1 +/- 0.5). CONCLUSION: Noninvasive quantitative measurements of myocardial perfusion in small animals at rest and during stress are feasible using PET. Evaluation of the coronary reserve must take into account the initial state of the anesthetized animal. The coronary reserve could be measured with both anesthetic agents using either dobutamine or adenosine stimulation. PMID- 15534065 TI - Uptake of 18F-fluorocholine, 18F-fluoroethyl-L-tyrosine, and 18F-FDG in acute cerebral radiation injury in the rat: implications for separation of radiation necrosis from tumor recurrence. AB - Differentiation between posttherapy radiation necrosis and recurrent tumor in humans with brain tumor is still a difficult diagnostic task. The new PET tracers (18)F-fluoro-ethyl-l-tyrosine (FET) and (18)F-fluorocholine (N,N-dimethyl-N-(18)F fluoromethyl-2-hydroxyethylammonium [FCH]) have shown promise for improving diagnostic accuracy. This study assessed uptake of these tracers in experimental radiation injury. METHODS: In a first model, circumscribed lesions were induced in the cortex of 35 rats using proton irradiation of 150 or 250 Gy. After radiation injury developed, uptake of (18)F-FET, (18)F-FCH, and (18)F-FDG was measured using autoradiography and correlated with histology and disruption of the blood-brain barrier as determined with Evans blue. In a second model, uptake of the tracers was assessed in acute cryolesions, which are characterized by the absence of inflammatory cells. RESULTS: Mean (18)F-FET, (18)F-FCH, and (18)F-FDG standardized uptake values in the most active part of the radiation lesion and the contralateral normal cortex (in parentheses) were 2.27 +/- 0.46 (1.42 +/- 0.23), 2.52 +/- 0.42 (0.61 +/- 0.12), and 6.21 +/- 1.19 (4.35 +/- 0.47). The degree of uptake of (18)F-FCH and (18)F-FDG correlated with the density of macrophages. In cryolesions, (18)F-FET uptake was similar to that in radiation lesions, and (18)F-FCH uptake was significantly reduced. CONCLUSION: Comparison of tracer accumulation in cryolesions and radiation injuries demonstrates that (18)F-FET uptake is most likely due to a disruption of the blood-brain barrier alone, whereas (18)F-FCH is additionally trapped by macrophages. Uptake of both tracers in the radiation injuries is generally lower than the published uptake in tumors, suggesting that (18)F-FET and (18)F-FCH are promising tracers for separating radiation necrosis from tumor recurrence. However, the comparability of our data with the literature is limited by factors such as different species and acquisition protocols and modalities. Thus, more studies are needed to settle this issue. Nevertheless, (18)F-FCH and (18)F-FET seem superior to (18)F-FDG for this purpose. PMID- 15534066 TI - Tumor imaging with 2 sigma-receptor ligands, 18F-FE-SA5845 and 11C-SA4503: a feasibility study. AB - Our objective was to study 2 radioligands for visualization of sigma-receptors with PET. METHODS: Two radioligands-sigma(1)-selective (11)C-1-(3,4 dimethoxyphenethyl)-4-(3-phenylpropyl)piperazine ((11)C-SA4503) and nonsubtype selective 1-(4-2'-(18)F-fluoroethoxy-3-methoxyphenethyl)-4-(3-(4 fluorophenyl)propyl)piperazine ((18)F-FE-SA5845)-were evaluated for tumor imaging. RESULTS: Binding studies to rat glioma cells (C6) and human nonsmall cell lung cancer cells (N417) indicated interaction of (18)F-FE-SA5845 with 2 sites and interaction of (11)C-SA4503 with a single site. Specific binding of (18)F-FE-SA5845 was 93% +/- 2% and that of (11)C-SA4503 was 78% +/- 6% of the total cellular uptake of radioactivity. Uptake of the (18)F-labeled ligand, but not that of the (11)C-labeled ligand, appeared to be related to the growth phase of the cells. Biodistribution experiments in C6 tumor-bearing nude rats (Ham HSD RNU rnu) indicated tumor-to-plasma ratios of 13.3 for (11)C-SA4503 and 8.0 for (18)F-FE-SA5845 and tumor-to-muscle ratios of 5.0 for (11)C-SA4503 and 4.9 for (18)F-FE-SA5845, 60 min after injection, which were reduced to values ranging from 1.4 to 2.0 after pretreatment of animals with haloperidol (2 micromol/kg). Tumor uptake of (18)F-FE-SA5845 showed a negative correlation with tumor size (P < 0.0001), in contrast to that of (11)C-SA4503, suggesting that tissue binding of the former ligand is related to cellular proliferation. A study with (11)C-SA4503 in a human volunteer indicated high uptake in liver, kidney, and heart but relatively low background in thorax and lower abdomen. CONCLUSION: Both (18)F-FE SA5845 and (11)C-SA4503 demonstrate specific binding to sigma-receptors in vivo and may be useful for the detection of pulmonary and abdominal tumors. However, the (18)F-labeled compound may be better for tumor staging than the (11)C-labeled drug. PMID- 15534067 TI - Pharmacokinetic animal PET study of FK506 as a potent neuroprotective agent. AB - The immunosuppressive agent FK506 (tacrolimus) has neuroprotective properties not only in rodents but also in nonhuman primates. To improve the accuracy of clinical studies of acute stroke, clinical dose setting based on brain concentrations of agents in humans is very helpful. We have already established a rapid-synthesis method for (11)C-labeled FK506; therefore, in the present study, we aimed to establish a method to measure brain concentrations of FK506 using (11)C-FK506 PET in monkeys. METHODS: Studies were performed on 3 male cynomolgus monkeys (Macaca fascicularis). FK506 (0.1 mg/kg) containing (11)C-FK506 was intravenously injected into the monkeys, and dynamic PET images were acquired for 30 min afterward. Arterial blood samples were collected 5 and 15 min after injection, and their radioactivities were measured by a gamma-counter. FK506 concentrations in brain and blood were calculated in units of moles per liter using the specific activity of the injected FK506. The PET study data were validated using an enzyme-linked immunosorbent assay. RESULTS: Seven minutes after administration, the radioactivity in the brain became constant and was maintained up to 30 min. We succeeded in measuring the FK506 concentration in the brain using (11)C-FK506 PET. Fifteen minutes after FK506 (0.1 mg/kg) administration, the concentrations in the cortex and striatum were 20.0 +/- 1.7 ng/g and 14.1 +/- 1.7 ng/g, respectively. FK506 concentrations in the blood correlated significantly with those measured by enzyme-linked immunosorbent assay. CONCLUSION: We successfully measured FK506 concentrations in anesthetized monkey brain and blood using (11)C-FK506 PET. These results indicate a potential method to measure FK506 concentrations in human brain. Additionally, a potential use for the PET technique in drug development has been demonstrated. PMID- 15534068 TI - Kinetic analysis of 18F-fluorodihydrorotenone as a deposited myocardial flow tracer: comparison to 201Tl. AB - The goals of this investigation were to assess the accuracy of (18)F fluorodihydrorotenone ((18)F-FDHR) as a new deposited myocardial flow tracer and to compare the results to those for (201)Tl. METHODS: The kinetics of these flow tracers in 22 isolated, erythrocyte- and albumin-perfused rabbit hearts were evaluated over a flow range encountered in patients. The 2 flow tracers plus a vascular reference tracer ((131)I-albumin) were introduced as a bolus through a port just above the aortic cannula. Myocardial extraction, retention, washout, and uptake parameters were computed from the venous outflow curves with the multiple-indicator dilution technique and spectral analysis. RESULTS: The mean +/ SD initial extraction fractions for (18)F-FDHR (0.85 +/- 0.07) and (201)Tl (0.87 +/- 0.05) were not significantly different, although the initial extraction fraction for (18)F-FDHR declined with flow (P < 0.0001), whereas the initial extraction fraction for (201)Tl did not. The washout of (201)Tl was faster (P < 0.001) and more affected by flow (P < 0.05) than was the washout of (18)F-FDHR. Except for the initial extraction fraction, (18)F-FDHR retention was higher (P < 0.001) and less affected by flow (P < 0.05) than was (201)Tl retention. Reflecting its superior retention, the net uptake of (18)F-FDHR was better correlated with flow than was that of (201)Tl at both 1 and 15 min after tracer introduction (P < 0.0001 for both comparisons). CONCLUSION: The superior correlation of (18)F-FDHR uptake with flow indicates that it is a better flow tracer than (201)Tl in the isolated rabbit heart. Compared with the other currently available positron-emitting flow tracers ((82)Rb, (13)N-ammonia, and (15)O-water), (18)F-FDHR has the potential of providing excellent image resolution without the need for an on-site cyclotron. PMID- 15534069 TI - Evaluation of parameters influencing S values in mouse dosimetry. AB - Clinical radionuclide therapy studies are commonly preceded by studies with small animals. Reliable evaluation of therapeutic efficacy must be based on accurate dosimetry. This study was performed to evaluate the influence of the mass of organs, the shape of organs, and the distances between organs on S values for mice. METHODS: A voxel-based version of a geometric model of a mouse was developed for input in our Monte Carlo program based on EGS4. Simulations were made for each source organ separately to resolve the S values for each organ. For verification purposes, S values were calculated for spheres of different masses and compared with the S values in the MIRDOSE3.1 software and with the S values on the Radiation Dose Assessment Resource Web site. The variation in the mass of the organs was determined from dissected mice. The influence of the shape of an organ was investigated by successive elongation of a sphere into spheroids with a constant mass. The right kidney was moved in the phantom of the mouse to evaluate the effect of organ distances on S values. The absorbed fractions for the mouse model presented here were compared with the results from some previously published models. The radionuclides used were (90)Y, (131)I, (111)In, and (99m)Tc. RESULTS: The results showed that the organ mass for one animal can differ by up to 33% from the mean mass. If linear interpolation from S value tables is used to obtain an S value for the specific mass of an organ, then the S value can differ by up to 80% from its true value. The corresponding deviation obtained by scaling according to mass is 20%. The shape of an organ was found to be the least important parameter for the S value. The cross-absorbed S value is strongly dependent on the geometry and the emitted radiation. For example, a 9.2 mm movement of the kidney can cause the S value from the liver to the right kidney to decrease to 0.05% of its original value for (90)Y. CONCLUSION: We conclude that the mass and the shape of organs and their locations relative to each other have considerable effects on mouse dosimetry. PMID- 15534070 TI - PET-based radiation dosimetry in man of 18F-fluorodihydrotestosterone, a new radiotracer for imaging prostate cancer. AB - 16 beta-fluoro-5 alpha-dihydrotestosterone (FDHT) is a promising new PET radiopharmaceutical for the imaging of prostate cancer. A recent clinical trial provided the opportunity for refinement of normal-tissue radiation-absorbed dose estimates based on quantitative PET. The objective of the current study was to derive estimates of normal-tissue absorbed doses for (18)F-FDHT administered to patients with advanced prostate cancer. METHODS: Absorbed dose estimates were derived from 10 (18)F-FDHT PET studies (administered activity, 111-407 MBq) of 7 prostate cancer patients. Activity concentrations in plasma and red marrow (assuming a plasmacrit of 0.58, an extracellular fluid fraction of 0.40, and equilibration of activity between plasma and marrow extracellular fluid) were measured ex vivo from a peripheral blood sample. Liver, spleen, urinary bladder contents, and total-body activities were measured by region-of-interest analysis of quantitative whole-body studies acquired with a dedicated PET scanner. Total organ activities and residence times were calculated from the respective PET scan derived activity concentrations assuming standard (70 kg) man organ masses. Urinary excretion was corrected for hepatobiliary excretion (liver activity), and a first-order adjustment was made for the bladder-wall mass based on the patient's total-body mass. Mean organ absorbed doses were calculated with the MIRD formalism and the standard man model using the MIRDOSE3 software program. RESULTS: The absorbed doses (mean +/- SD) ranged from 0.00057 +/- 0.000281 cGy/MBq (to skin) to 0.00868 +/- 0.00481 cGy/MBq (to bladder wall) (voiding intervals, 1-2 h), and the effective dose equivalent was 0.00177 +/- 0.000152 cSv/MBq. CONCLUSION: The maximum absorbed dose among all tissues in all 10 studies, 0.0151 cGy/MBq, occurred for the urinary bladder wall (with hydration and 1- to 2-h voiding intervals). To ensure that the maximum normal-tissue absorbed dose is kept below the recommended maximum permissible dose of 5 cGy per single administration, a maximum administered activity of 331 MBq (5 cGy/[0.0151 cGy/MBq]) is recommended for (18)F-FDHT. PMID- 15534071 TI - Re: Fischman and Thrall editorial, "Who Should Read ... PET Studies". PMID- 15534072 TI - Diagnostic Value of PET in Cardiac Sarcoidosis. PMID- 15534073 TI - Angiotensin subtype-2 receptors inhibit renin biosynthesis and angiotensin II formation. AB - Renin is regulated by angiotensin subtype 1 (AT1) receptor, but it is unknown whether angiotensin subtype 2 (AT2) receptor contributes to this regulation. We hypothesized that AT2 receptors inhibit angiotensin II (Ang II) through inhibition of renin biosynthesis. We monitored changes in renal Ang II, renin mRNA and protein expression, and plasma renin concentration (PRC) in response to renal cortical administration of the AT1 receptor blocker valsartan or the AT2 receptor blocker PD 123319 (PD) in conscious rats administered low sodium intake (LS). Renal interstitial Ang II increased by 47-fold in response to LS and increased further in response to valsartan or PD by 67-fold and 61-fold from normal sodium diet (NS) and by 41% and 29% from LS, respectively. Renin mRNA increased 63% during LS, and either valsartan or PD increased it further by 600% and 250% from NS and 538% and 187% from LS, respectively. Similarly, renal renin content and PRC increased in response to LS and increased further in response to combined LS and valsartan or PD administration. Immunostaining for renal renin protein demonstrated an increase in its expression in juxtaglomular and tubular cells during LS and increased further during AT1 or AT2 receptor blockade. These data demonstrate for the first time to our knowledge that AT2 receptors regulate the renin-angiotensin system activity via inhibition of renin synthesis. PMID- 15534074 TI - Genome-wide scan for blood pressure suggests linkage to chromosome 11, and replication of loci on 16, 17, and 22. AB - Hypertension was one of the first complex traits to be studied and is thought to be influenced by polygenic and multiple environmental risk factors. Several genomic studies have found suggestive logarithm of odds (LOD) scores for either blood pressure or essential hypertension, but few loci have been replicated. In this study, we performed a genome-wide linkage analysis for systolic blood pressure (SBP) and diastolic blood pressure (DBP) on 1109 white female dizygotic twin pairs from the TwinsUK registry in London. Multipoint linkage analysis replicated the locations of 3 previously reported linkage peaks: on chromosome 16 at 65 cM (LOD 0.8 for SBP and 1.8 for DBP); on chromosome 17 at 70 cM (LOD 1.8 SBP); and at 35 cM on chromosome 22 (LOD 0.97 SBP and 0.99 DBP). Results from multipoint analysis showed 1 novel suggestive linkage for SBP (multipoint LOD 2.28; 2-point P=0.0007) at 35 cM on chromosome 11. Results were similar when those on blood pressure medication were excluded. These are encouraging results for hypertensive research and demonstrate that despite past disappointments, linkage studies can be used to replicate regions from other studies and potentially discover new genetic risk factors of moderate to large effect size. Considering the differences in selection and ascertainment of the previous linkage studies, these results also suggest that some quantitative trait loci are likely to influence the normal range of blood pressure and clinical hypertension, whereas others will be specific to each trait. Future studies should focus on the fine mapping of these replicated regions, which include potential candidate genes. PMID- 15534075 TI - Arterial stiffness and wave reflections in patients with sickle cell disease. AB - We tested the hypothesis that lower blood pressure and increased vasodilatation reported in sickle cell disease (SCD) patients with hemoglobin SS genotype (SS) are translated by lower arterial stiffness determined by pulse wave velocity (PWV) and wave reflections assessed by augmentation index (AI). We enrolled 20 SS (8 females; 12 male) patients closely matched for age, gender, height, and body mass index to 20 subjects with hemoglobin AA genotype (AA). Carotid-femoral PWV (PWV(CF)) and carotid-radial PWV (PWV(CR)) were recorded with the Complior device. Aortic AI was derived from pressure wave analysis (SphygmocoR). PWV(CF) and PWV(CR) were lower in SS than in AA (4.5+/-0.7 m/s versus 6.9+/-0.9 m/s, P<0.0001 and 6.6+/-1.2 m/s versus 9.5+/-1.4 m/s, P<0.0001, respectively). AI was lower in SS than in AA (2+/-14% versus 11+/-8%, P=0.02). Multivariate analysis revealed that both PWV(CF) and PWV(CR) were negatively associated with hemoglobin SS type and positively related to mean arterial pressure (MAP), whereas AI was positively associated with MAP and total cholesterol (all P<0.0001). Multivariate analysis restricted to SS indicated a positive association between PWV(CF) and PWV(CR) with age but a negative association with MAP (R2=0.57 and 0.51, respectively, both P<0.001), whereas MAP and heart rate were independently associated with AI (R2=0.65, P<0.001). This study provides the first evidence that SCD is associated with both lower arterial stiffness and wave reflections. SS patients have a paradoxical negative association between PWV and MAP, suggesting that low MAP does not protect them against arterial stiffness impairment. PMID- 15534076 TI - Job strain, ethnicity, and sympathetic nervous system activity. AB - Several studies have demonstrated that blacks have heightened pressor sensitivity in response to the alpha-agonist, phenylephrine. However, studies examining whether psychosocial factors contribute to this difference are scarce. We examined the effects of job strain on pressor sensitivity in 76 whites and 46 blacks who were enrolled in a study of stress, sleep, and blood pressure. Responses to phenylephrine were examined at an inpatient clinical research center. After a 3-minute baseline period, a 100-microgram phenylephrine bolus was administered to participants intravenously. To measure catecholamines, 24-hour urine samples were also collected from participants. There was a significant relationship between job strain and pressor sensitivity, such that individuals with low decisional control and high job demands experienced a greater increase in diastolic pressure after receiving phenylephrine. Low decisional control was also associated with decreased baroreflex sensitivity. There was an interaction between ethnicity and job control on blood pressure responses to phenylephrine and on 24-hour urinary norepinephrine levels. Blacks who perceived less control experienced a greater increase in diastolic pressure after receiving phenylephrine and had elevated norepinephrine levels. These findings suggest possible mechanisms by which job strain may be associated with cardiovascular disease. PMID- 15534077 TI - Systolic blood pressure during recovery from exercise and the risk of acute myocardial infarction in middle-aged men. AB - We prospectively assessed the association of systolic blood pressure (SBP) after exercise with the risk of an acute myocardial infarction. Limited information exists currently on the role of SBP during recovery period with the risk of acute myocardial infarction. SBP was measured every 2 minutes during and after a progressive cycle ergometer exercise test in a representative sample of 2336 men (aged 42 to 61 years). During an average follow-up period of 13.1 years, 358 acute myocardial infarctions occurred. An incremental rise of 10 mm Hg per minute in SBP at 2 minutes after exercise (relative risk, 1.07-fold; 95% confidence interval [CI], 1.03 to 1.12; P=0.001) was associated with the risk of acute myocardial infarction after adjustment for age, alcohol consumption, smoking, serum lipids, diabetes mellitus, body mass index, resting SBP, regular use of antihypertensive medications, physical fitness, heart rate, and ischemic ECG findings during exercise. Men with elevated SBP of >195 mm Hg after exercise had a 1.69-fold (95% CI, 1.24 to 2.30; P=0.001) risk for an acute myocardial infarction compared with those with SBP <170 mm Hg after adjustment for age, other risk factors, and resting SBP. SBP after exercise provides an incremental predictive value for acute myocardial infarction beyond that of resting SBP. This emphasizes the importance of SBP measurements after the exercise test because it provides additional valuable prognostic measure with regard to acute myocardial infarction. PMID- 15534078 TI - Osteopontin modulates myocardial hypertrophy in response to chronic pressure overload in mice. AB - Osteopontin (OPN) expression increases in the heart during hypertrophy and heart failure. Here, we studied the role of OPN in pressure overload-induced hypertrophy and analyzed the signaling pathways involved in hypertrophy. Aortic banding (AB) was performed in a group of wild-type (WT) and OPN knockout (KO) mice to induce pressure overload. Left ventricular (LV) structural and functional remodeling was studied 1 month after AB. AB increased OPN and beta1 integrin (a receptor for OPN) protein expression in WT-AB group. Hypertrophic response as measured by increased heart weight-to-body weight ratio and myocyte cross sectional area was significantly increased in WT-AB and KO-AB groups when compared with their respective shams. However, the increase was significantly higher in WT-AB. Re-expression of atrial natriuretic factor was only detected in WT-AB group. LV end-diastolic pressure-volume curve obtained using Langendorff perfusion analysis exhibited a leftward shift in WT-AB group, not in KO-AB. LV developed pressures measured over a range of LV volumes were significantly increased in WT-AB, not in KO-AB mice. Increased phosphorylation of c-Jun N terminal kinases, p38 kinase, Akt, and glycogen synthase kinase-3beta was significantly higher in WT-AB when compared with KO-AB group. Increased OPN expression may play an essential role in modulating compensatory cardiac hypertrophy in response to chronic pressure overload. PMID- 15534079 TI - Back to the basic: measuring QRS voltage-interval in left ventricular hypertrophy. PMID- 15534080 TI - Imaging molecular signatures in oligodendroglioma. PMID- 15534081 TI - Administration of CI-1033, an irreversible pan-erbB tyrosine kinase inhibitor, is feasible on a 7-day on, 7-day off schedule: a phase I pharmacokinetic and food effect study. AB - PURPOSE: To determine the maximum tolerated dose of administrating CI-1033, an oral 4-anilinoquinazoline that irreversibly inhibits the tyrosine kinase domain of all erbB subfamilies, on an intermittent schedule, and assess the interaction of CI-1033 with food on the pharmacokinetic behavior. EXPERIMENTAL DESIGN: Escalating doses of CI-1033 from a dose level of 300 mg/day for 7 days every other week were administered to patients with advanced solid malignancies. Plasma concentration-time data sets from all evaluable patients were used to develop a population pharmacokinetic model. Noncompartmental methods were used to independently assess the effect of a high-fat meal on CI-1033 absorption and bioavailability. RESULTS: Twenty-four patients were treated with 69 twenty-eight day courses. The incidence of unacceptable toxicity, principally diarrhea and skin rash, was observed at the 300 mg/day dose level. At the 250 mg/day level, toxicity was manageable, and protracted administration was feasible. A one compartment linear model with first-order absorption and elimination adequately described the pharmacokinetic disposition. CL/F, apparent volume of distribution (Vd/F), and ka (mean +/- relative SD) were 280 L/hour +/- 33%, 684 L +/- 20%, and 0.35 hour(-1)+/- 69%, respectively. Cmax values were achieved in 2 to 4 hours. Systemic CI-1033 exposure was largely unaffected by administration of a high-fat meal. At 250 mg, concentration values exceeded IC50 values required for prolonged pan-erbB tyrosine kinase inhibition in preclinical assays. CONCLUSIONS: The recommended dose on this schedule is 250 mg/day. Its tolerability and the biological relevance of concentrations achieved at the maximal tolerated dose warrant consideration of disease-directed evaluations. This intermittent treatment schedule can be used without regard to meals. PMID- 15534082 TI - The influence of androgen deprivation therapy on dihydrotestosterone levels in the prostatic tissue of patients with prostate cancer. AB - PURPOSE: The influence of androgen deprivation therapy on dihydrotestosterone levels in the prostatic tissue is not clearly known. Changes in dihydrotestosterone levels in the prostatic tissue during androgen deprivation therapy in the same patients have not been reported. We analyzed dihydrotestosterone levels in prostatic tissue before and after androgen deprivation therapy. EXPERIMENTAL DESIGN: A total of 103 patients who were suspected of having prostate cancer underwent prostatic biopsy. Sixty-nine patients were diagnosed as having prostate cancer whereas the remaining 34 were negative. Serum samples were collected before biopsy or prostatectomy. Dihydrotestosterone levels in prostatic tissue and serum were analyzed using liquid chromatography/electrospray ionization-mass spectrometry after polar derivatization. In 30 of the patients with prostate cancer, dihydrotestosterone levels in prostatic tissue were determined by performing rebiopsy or with prostate tissues excised after 6 months on androgen deprivation therapy with castration and flutamide. RESULTS: Dihydrotestosterone levels in prostate tissue after androgen deprivation therapy remained at approximately 25% of the amount measured before androgen deprivation therapy. Dihydrotestosterone levels in serum decreased to approximately 7.5% after androgen deprivation therapy. The level of dihydrotestosterone in prostatic tissue before androgen deprivation therapy was not correlated with the serum level of testosterone. Serum levels of adrenal androgens were reduced to approximately 60% after androgen deprivation therapy. CONCLUSIONS: The source of dihydrotestosterone in prostatic tissue after androgen deprivation therapy involves intracrine production within the prostate, converting adrenal androgens to dihydrotestosterone. Dihydrotestosterone still remaining in prostate tissue after androgen deprivation therapy may require new therapies such as treatment with a combination of 5alpha-reductase inhibitors and antiandrogens, as well as castration. PMID- 15534083 TI - A phase I open label study of the farnesyltransferase inhibitor CP-609,754 in patients with advanced malignant tumors. AB - PURPOSE: The purpose of this phase I clinical trial was to determine the maximum tolerated dose and toxicity of CP-609,754 in patients with solid tumors refractory to standard therapies, to determine the cellular effects of CP-609,754 on its molecular target (farnesyltransferase), and to determine the recommended phase II dose (RP2D) of this agent. EXPERIMENTAL DESIGN: Consenting patients with adequate bone marrow, liver, and renal function were enrolled with an accelerated dose strategy with single-patient parallel cohorts in whom the drug was given orally either once or twice daily. Once a dose-limiting toxicity was encountered or two patients developed Common Toxicity Criteria > or = grade 2 toxicities, a modified Fibonacci sequence was initiated. Blood samples were collected during cycle 1 for pharmacokinetic and pharmacodynamic analyses. RESULTS: A total of 68 cycles of CP-609,754 was administered to 21 patients enrolled in this study. The dose escalation was from 20 mg once daily to 640 mg twice per day, and at the highest dose level, one of six patients developed a dose-limiting toxicity of grade 3 neuropathy. The drug was otherwise well tolerated, and the maximum tolerated dose was not reached because of the large number of tablets that would have been required for additional dose escalation. Pharmacokinetic analyses showed a proportional increase in exposure with dose, rapid oral absorption, and a half-life of approximately 3 hours. Pharmacodynamic results predict a 95% maximal inhibition of peripheral blood mononuclear cell farnesyltransferase activity 2 hours postdose, on average, with a dose of 400 mg twice per day of CP 609,754. CONCLUSIONS: On the basis of the safety findings and the pharmacokinetic and pharmacodynamic analyses, the RP2D of CP-609,754 is > or =640 mg twice per day. PMID- 15534084 TI - Phase II feasibility and pharmacokinetic study of concurrent administration of trastuzumab and high-dose chemotherapy in advanced HER2+ breast cancer. AB - PURPOSE: To evaluate the safety of concurrent treatment with trastuzumab and high dose chemotherapy (HDC), using cyclophosphamide, cisplatin, and 1,3-bis(2 chloroethyl)-1-nitrosourea (BCNU), with autologous hematopoietic progenitor cells support, in patients with HER2+ advanced breast cancer. EXPERIMENTAL DESIGN: Patients with HER2-overexpressing high-risk primary breast cancer (HRPBC; defined as > or =4 involved nodes or inflammatory disease), or metastatic breast cancer (MBC) were eligible. Treatment consisted of a loading dose of trastuzumab at 4 mg/kg (day -5), HDC (days -5 to -2), autologous hematopoietic progenitor cells infusion on day 0, and weekly maintenance trastuzumab (2 mg/kg) from day +1 (minimum of 9 doses). Cardiac monitoring included serial left ventricular ejection fraction measurements before treatment and on days +20 and +65. RESULTS: Thirty-three patients were prospectively enrolled (13 HRPBC, 20 MBC). Toxicity seemed similar to that expected with this HDC regimen alone. Neutrophils and platelets engrafted promptly. There were no cases of grade 4 or 5 toxicity. One patient experienced symptomatic grade 3 acute cardiac failure on day -4, responsive to treatment. Trastuzumab did not alter the pharmacokinetics of HDC. Eleven of twelve MBC patients with measurable disease (nine of them refractory to previous chemotherapy) experienced an objective response (9 complete and 2 partial responses). At median follow-up of 34 (13-58) months, all HRPBC patients remain alive and free of disease; the MBC group has event-free survival and overall survival rates of 45 and 70%, respectively. CONCLUSIONS: Incorporation of trastuzumab into HDC (cyclophosphamide, cisplatin, and BCNU) is feasible, with no apparent increased toxicity or pharmacokinetic interactions. PMID- 15534085 TI - High LYVE-1-positive lymphatic vessel numbers are associated with poor outcome in breast cancer. AB - PURPOSE: The clinical significance of intratumoral or peritumoral lymph vessel density is not known. LYVE-1, a lymphatic endothelium-specific hyaluronan receptor, is a novel lymphatic vessel marker that is expressed on lymph vessel endothelial cells of both normal and neoplastic tissues. EXPERIMENTAL DESIGN: We investigated expression of LYVE-1 by immunohistochemistry in 180 unilateral, invasive ductal breast carcinomas and assessed the presence and density of lymph vessels within the tumor and at the tumor periphery. RESULTS: A minority (12%) of breast carcinomas had intratumoral lymph vessels, whereas peritumoral lymph vessels were identified in almost all cases (94%). No substantial association was found between the number of LYVE-1-positive vessels and the number of CD31 or vascular endothelial growth factor receptor-3-positive vessels, or vascular endothelial growth factor-C expression. The number of metastatic axillary lymph nodes increased in parallel with increasing lymph vessel counts (P = 0.033). A higher than the median lymph vessel count at the tumor periphery was significantly associated with unfavorable distant disease-free survival and overall survival. Women with high peritumoral lymph vessel density had only 58% (95% confidence interval, 46-70%) 5-year distant disease-free survival as compared with 74% (66-83%) among those with a low peritumoral lymph vessel density (P = 0.0088). In contrast, the presence of intratumoral lymph vessels was associated with neither axillary nodal status nor survival. Lymph vessel density was not an independent prognostic factor in a multivariate survival analysis. CONCLUSIONS: A high peritumoral lymph vessel density is associated with a poor outcome in ductal breast cancer. PMID- 15534086 TI - Genetic analysis of the RNASEL gene in hereditary, familial, and sporadic prostate cancer. AB - PURPOSE: The RNASEL gene has been proposed as a candidate gene for the HPC1 locus through a positional cloning and candidate gene approach. Cosegregation between the truncating mutation E265X and disease in a hereditary prostate cancer (HPC) family and association between prostate cancer risk and the common missense variant R462Q has been reported. To additionally evaluate the possible role of RNASEL in susceptibility to prostate cancer risk, we performed a comprehensive genetic analysis of sequence variants in RNASEL in the Swedish population. EXPERIMENTAL DESIGN: Using 1624 prostate cancer cases and 801 unaffected controls, the truncating mutation E265X and five common sequence variants, including the two missense mutations R462Q and D541E, were evaluated for association between genotypes/haplotypes and prostate cancer risk. RESULTS: The prevalence of E265X carriers among unaffected controls and prostate cancer patients was almost identical (1.9 and 1.8% in controls and cases, respectively), and evidence for segregation of E265X with disease was not observed within any HPC family. Overall, the analyses of common sequence variants provided limited evidence for association with prostate cancer risk. We found a marginally significant inverse association between the missense mutation D541E and sporadic prostate cancer risk (odds ratio, 0.77; 95% confidence interval, 0.59-1.00) and reduced risk of prostate cancer in carriers of two different haplotypes being completely discordant. CONCLUSIONS: Considering the high quality in genotyping and the size of this study, these results provide solid evidence against a major role of RNASEL in prostate cancer etiology in Sweden. PMID- 15534087 TI - Increased serum interleukin-8 in patients with early and metastatic breast cancer correlates with early dissemination and survival. AB - PURPOSE: The prognostic significance of serum interleukin (IL)-8 was evaluated in patients with metastatic breast cancer. The predictive value of serum IL-8 for the presence of occult metastatic tumor cells in bone marrow aspirates was evaluated in patients with operable and metastatic breast cancer. EXPERIMENTAL DESIGN: Serum IL-8 was measured in healthy controls, patients with operable breast cancer, and patients with untreated, progressive metastatic breast cancer. In 69 patients with either operable or advanced breast cancer, occult cytokeratin positive cells were counted in bone marrow aspirates. RESULTS: Serum IL-8 levels are increased in 67% (52 of 77) of patients with advanced breast cancer. Overall, these levels are significantly higher in patients with breast cancer compared with healthy volunteers (P < 0.001). The IL-8 levels increase significantly in patients with more advanced disease. An elevated serum IL-8 is related to an accelerated clinical course, a higher tumor load, and the presence of liver or lymph node involvement. A multivariate analysis indicates that serum IL-8 is an independent significant factor for postrelapse survival. There was a significant difference between serum IL-8 levels in patients with or without occult cytokeratin-positive bone marrow cells (P < 0.04). Serum IL-8 levels also showed an association with the number of these cells (P < 0.01). CONCLUSIONS: Serum IL-8 is increased in patients with breast cancer and has an independent prognostic significance for postrelapse survival. The observations on the relationship between occult cytokeratin-positive bone marrow cells corroborate the concept of IL-8 acting as a contributor to the process of tumor cell dissemination. Similarly, the relationship between serum IL-8 and nodal stage at presentation deserves further study. These results further expand the concept that inflammation and inflammatory cytokines are critical components of tumor progression. PMID- 15534088 TI - Delineation of brain tumor extent with [11C]L-methionine positron emission tomography: local comparison with stereotactic histopathology. AB - PURPOSE: Methyl-[11C]L-methionine ([11C]MET) positron emission tomography (PET) in brain tumors reflects amino acid transport and has been shown to be more sensitive than magnetic resonance imaging in stereotactic biopsy planning. It remains unclear whether the increased [11C]MET uptake is limited to solid tumor tissue or even detects infiltrating tumor parts. EXPERIMENTAL DESIGN: In 30 patients, a primary or recurrent brain tumor was suspected on magnetic resonance imaging. Patients were investigated with [11C]MET-PET before stereotactic biopsy. The biopsy trajectories were plotted into the [11C]MET-PET images with a newly designed C-based software program. The exact local [11C]MET uptake was determined within rectangular regions of interest of 4 mm in width and length aligned with the biopsy specimen. Individual histologic specimens were rated for the presence of solid tumor tissue, infiltration area, and nontumorous tissue changes. RESULTS: Receiver operating characteristics analysis demonstrated a sensitivity of 87% and specificity of 89% for the detection of tumor tissue at a threshold of 1.3-fold [11C]MET uptake relative to normal brain tissue. At this threshold, only 13 of 100 tumor positive specimen were false negative mainly in grade 2 astrocytoma. In grade 2 astrocytoma, mean [11C]MET uptake in the infiltration area was significantly higher than in solid tumor tissue (P < 0.003). CONCLUSIONS: [11C]MET-PET detects solid parts of brain tumors, as well as the infiltration area at high sensitivity and specificity. High [11C]MET uptake in infiltrating tumor of astrocytoma WHO grade 2 reflects high activity in this tumor compartment. Molecular imaging, with [11C]MET, will guide improved management of patients with brain tumors. PMID- 15534089 TI - Angiogenic profile of breast carcinoma determines leukocyte infiltration. AB - To study the relationship between the angiogenic profile and leukocyte infiltration of tumors, single cell suspensions of archival frozen medullary and ductal breast cancer tissues were analyzed by flow cytometry. The amount of leukocytes and endothelial cells was measured, as well as the expression of intercellular adhesion molecule-1 (ICAM-1) on the endothelial cell fraction. A significantly higher number (3.2-fold) of infiltrating leukocytes was observed in medullary carcinoma. The composition of this infiltrate was similar to that seen in ductal carcinomas. The more intense infiltrate was explained by the approximately 3-fold enhanced endothelial ICAM-1 expression in medullary carcinoma. The angiogenic profile of all tumors was assessed by quantitative real time reverse transcription-PCR analysis. Vascular endothelial growth factor (VEGF)-C and VEGF-D, but not VEGF-A, basic fibroblast growth factor, placental growth factor, and angiopoietins 1, 2, and 3 showed a relatively higher level of expression in ductal carcinoma than in medullary carcinoma. In vitro, both VEGF-C and VEGF-D were found to decrease endothelial ICAM-1 expression in the presence of basic fibroblast growth factor. These data suggest that in vivo angiogenic stimuli prevent the formation of an effective leukocyte infiltrate in tumors by suppressing endothelial ICAM-1 expression. PMID- 15534090 TI - Monoclonal immunoglobulin production is a frequent event in patients with mucosa associated lymphoid tissue lymphoma. AB - PURPOSE: Mucosa-associated lymphoid tissue (MALT) lymphoma comprises 7% of all newly diagnosed non-Hodgkin's lymphomas and is therefore among the most common lymphoma entities. Monoclonal gammopathy due to production of a monoclonal immunoglobulin by lymphoma cells is a well-known phenomenon associated with various types of B-cell non-Hodgkin's lymphomas. The objective of the present study was to evaluate the incidence and clinical relevance of paraprotein (PP) production in patients with MALT lymphoma. EXPERIMENTAL DESIGN: Fifty two patients were prospectively evaluated with regard to differentiation of the MALT lymphoma cells, t(11;18) translocation, monoclonal immunoglobulin production, Helicobacter pylori (HP) status, stage, treatment, and clinical outcome. RESULTS: Nineteen of 52 MALT lymphoma patients (36%) had PP (8 IgMkappa, 6 IgGkappa, 4 IgMlambda, and 1 IgAkappa). The histologic feature of plasmacytic differentiation correlated significantly with the production of PP (P = 0.001). No correlation was found between PP and clinical stage, HP status, and t(11;18) status. PP was, however, negatively associated with response to eradication of HP in gastric MALT lymphoma, and PP levels declined significantly in patients responding to chemotherapy or radiation. Importantly, both immunofixation and serum electrophoresis have to be performed to detect low PP levels. CONCLUSIONS: In conclusion, PP levels may probably be used as a potential prognostic tool for response to HP eradication, and serial measurements may also allow for noninvasive assessment of response to radiation or chemotherapy in patients with MALT lymphoma. PMID- 15534091 TI - Correlation of molecular genetics with molecular and morphological imaging in gliomas with an oligodendroglial component. AB - PURPOSE: Since the recognition that oligodendrogliomas may be chemosensitive, their diagnosis and clinical management has become highly controversial. Histopathology diagnosis remains challenging and new tools such as molecular genetics or molecular imaging require evaluation. EXPERIMENTAL DESIGN: In a single-center, population-based prospective study, allelic imbalance in chromosomes 1p36, 19q13, 17p13, 10p12-15, and 10q22-26 has been investigated in 19 oligodendroglioma WHO grade 2 (OII), 20 oligoastrocytoma WHO grade 2 (OAII), 8 oligodendroglioma WHO grade 3 (OIII), and 12 oligoastrocytoma WHO grade 3 (OAIII), and compared with pretherapy histopathology, computed tomography and/or magnetic resonance (CT and/or MR), [fluorine-18]fluoro-2-deoxyglucose (18F-FDG), and thallium-201 single-photon emission computed tomography (201Tl SPECT). RESULTS: In 50 cases, 18F-FDG uptake correlated with 201Tl uptake; however, 8 cases had increased 201Tl uptake but were hypometabolic for 18F-FDG, and 1 case was hypermetabolic with normal 201Tl uptake. Sixteen cases enhanced on CT/MR but failed to show 201Tl uptake; and 2 low-grade non-enhancing oligodendrogliomas had increased 201Tl uptake. Increased metabolism was more likely in high-grade cases, with 201Tl uptake more strongly correlated with grade than was 18F-FDG uptake. Tumors with 1p/19q loss were more likely to show increased 201Tl uptake and, to a lesser degree, increased 18F-FDG uptake than those without these losses. Elevated metabolism in 28% of low-grade tumors was significantly more common in tumors with 1p/19q loss, and increased uptake of both 18F-FDG and 201Tl in low-grade cases was found only in those with 1p/19q loss. CONCLUSIONS: In this study, dissociation of uptake of contrast agents and radiotracers suggests independent deregulation of the blood-brain barrier breakdown and metabolism during disease progression of oligodendroglial neoplasms, and the association of elevated metabolism with 1p/19q loss, particularly in low-grade tumors, may have implications for clinical management. PMID- 15534092 TI - 7-hydroxystaurosporine (UCN-01) inhibition of Akt Thr308 but not Ser473 phosphorylation: a basis for decreased insulin-stimulated glucose transport. AB - 7-hydroxystaurosporine (UCN-01) infused for 72 hours by continuous i.v. infusion induced insulin resistance during phase I clinical trials. To understand the mechanism for this observation, we examined the effect of UCN-01 on insulin stimulated glucose transport activity with 3-O-methylglucose in isolated rat adipose cells. UCN-01 inhibits glucose transport activity in a dose-dependent manner at all insulin concentrations. At the clinically relevant concentration of 0.25 mumol/L UCN-01, glucose transport is inhibited 66, 29, and 26% at insulin concentrations of 10, 50, and 100,000 (100K) microunits/mL respectively, thus shifting the dose-response curve to the right. Increasing concentrations of UCN 01 up to 2.5 mumol/L progressively shift the insulin dose-response curve even further. As Akt is known to mediate in part action initiated at the insulin receptor, we also studied the effect of UCN-01 on Akt activation in whole-cell homogenates of these cells. Decreased glucose transport activity directly parallels decreased Akt Thr308 phosphorylation in both an insulin and UCN-01 dose dependent manner, whereas Akt Ser473 phosphorylation is inhibited only at the lowest insulin concentration, and then, only modestly. UCN-01 also inhibits insulin-induced Thr308 but not Ser473 phosphorylation of Akt associated with the plasma membranes and low-density microsomes and inhibits translocation of GLUT4 from low-density microsomes to plasma membranes as expected from the glucose transport activity measurements. These data suggest that UCN-01 induces clinical insulin resistance by blocking Akt activation and subsequent GLUT4 translocation in response to insulin, and this effect appears to occur by inhibiting Thr308 phosphorylation even in the face of almost completely unaffected Ser473 phosphorylation. PMID- 15534093 TI - Impact of human neutralizing antibodies on antitumor efficacy of an oncolytic adenovirus in a murine model. AB - PURPOSE: The purpose of this study was to assess the impact of anti-adenovirus neutralizing antibodies (AdNAbs) on the distribution, tolerability, and efficacy of intravenously administered oncolytic adenovirus. A translational model was developed to evaluate the impact of humoral immunity on intravenous administration of oncolytic adenovirus in humans. EXPERIMENTAL DESIGN: Initially, severe combined immunodeficient (SCID)/beige mice were passively immunized with various amounts of human sera to establish a condition of preexisting humoral immunity similar to humans. A replication-deficient adenovirus encoding beta galactosidase (rAd-betagal) was injected intravenously into these mice. An AdNAb titer that mitigated galactosidase transgene expression was determined. A xenograft tumor-bearing nude mouse model was developed to assess how a similar in vivo titer would impact the activity of 01/PEME, an oncolytic adenovirus, after intravenous administration. RESULTS: In SCID/beige mice, there was a dose dependence between AdNAbs and galactosidase transgene expression; 90% of transgene expression was inhibited when the titer was 80. A similar titer reconstituted in the nude mice with human serum, as was done in the SCID/beige mice, did not abrogate the antitumor efficacy of the replicating adenovirus after intravenous administration. Viral DNA increased in tumors over time. CONCLUSIONS: In intravenous administration, preexisting AdNAb titer of 80 significantly attenuated the activity of a 2.5 x 10(12) particles per kilogram dose of nonreplicating adenovirus; the same titer had no affect on the activity of an equivalent dose of replicating adenovirus. Our results suggest that a majority of patients with preexisting adenovirus immunity would be candidates for intravenous administration of oncolytic adenovirus. PMID- 15534094 TI - In vitro stimulation with WT1 peptide-loaded Epstein-Barr virus-positive B cells elicits high frequencies of WT1 peptide-specific T cells with in vitro and in vivo tumoricidal activity. AB - The Wilms tumor protein (WT1) is overexpressed in most acute and chronic leukemias. To develop a practicable, clinically applicable approach for generation of WT1-specific T cells and to comparatively evaluate the immunogenicity of WT1 in normal individuals, we sensitized T cells from 13 HLA A0201+ and 5 HLA-A2402+ donors with autologous EBV-transformed B cells or cytokine-activated monocytes, loaded with the HLA-A0201-binding WT1 peptides (126 134)RMFPNAPYL or (187-195)SLGEQQYSV or a newly identified HLA-A2402-binding WT1 peptide (301-310)RVPGVAPTL. WT1-specific T cells were regularly generated from each donor. T cells sensitized with peptide-loaded EBV-transformed B cells generated higher numbers of WT1-specific T cells than peptide-loaded cytokine activated monocytes. Contrary to expectations, the frequencies of WT1 peptide specific T cells were equivalent to those generated against individual highly immunogenic HLA-A0201-binding EBV peptides. Each of these T-cell lines specifically killed WT1+ leukemias and solid tumors in an HLA-restricted manner but did not lyse autologous or HLA-matched normal CD34+ hematopoietic progenitor cells or reduce their yield of colony-forming unit-granulocyte-macrophage (CFU GM), burst-forming unit erythroid (BFU-E), or mixed colonies (CFU-mix). Furthermore, WT1 peptide-specific T cells after adoptive transfer into nonobese diabetic-severe combined immunodeficient mice bearing subcutaneous xenografts of WT1+ and WT1- HLA-A0201+ leukemias preferentially accumulated in and induced regressions of WT1+ leukemias that expressed the restricting HLA allele. Such cells are clinically applicable and may prove useful for adoptive cell therapy of WT1+ malignant diseases in humans. PMID- 15534095 TI - Phase I study of docetaxel in combination with the P-glycoprotein inhibitor, zosuquidar, in resistant malignancies. AB - PURPOSE: To determine the maximum tolerated dose, dose-limiting toxicity, and pharmacokinetics of docetaxel infused over 1 hour when given in combination with oral zosuquidar to patients with resistant solid tumors. EXPERIMENTAL DESIGN: In cycle 1, patients received docetaxel alone. In subsequent cycles, zosuquidar was administered with docetaxel, which was escalated from 75 to 100 mg/m2. Zosuquidar was escalated from 100 to 300 mg/m2 every 8 hours on days 1 to 3 for a total of 7 doses, or from 400 to 500 mg every 12 hours for 2 doses administered 2 hours before docetaxel. The pharmacokinetics of docetaxel with and without zosuquidar administration were obtained. RESULTS: Thirty-six of 41 patients completed at least one cycle of docetaxel and zosuquidar. The maximum tolerated dose was docetaxel 100 mg/m2 and zosuquidar 500 mg every 12 hours for 2 doses. The most common toxicity was neutropenia. In 35 patients, zosuquidar produced minimal increases in the docetaxel peak plasma concentrations and area under the curve. Dosing over 3 days with zosuquidar (7 doses) did not show benefit over the 1-day dosing. Of the 36 patients, one patient had a partial response, and 14 patients had disease stabilization. CONCLUSIONS: Docetaxel at 75 or 100 mg/m2 and zosuquidar 500 mg 2 hours before docetaxel and 12 hours later is well tolerated. Zosuquidar minimally alters the pharmacokinetics of docetaxel, allowing full dose docetaxel to be given with this P-glycoprotein modulator. A Phase II study with this combination in advanced breast carcinoma is underway. PMID- 15534096 TI - A phase I and pharmacokinetic study of exisulind and docetaxel in patients with advanced solid tumors. AB - PURPOSE: Exisulind (sulindac sulfone, FGN-1, Aptosyn) is a sulindac metabolite that induces apoptosis via inhibition of cyclic GMP-phosphodiesterase. This agent demonstrated tumor growth inhibition in rodent models of colon, breast, prostate, and lung carcinogenesis. In an orthotopic model of human non-small-cell lung cancer, the combination of exisulind and docetaxel prolonged survival in athymic nude rats, forming the basis of this phase I combination study. EXPERIMENTAL DESIGN: This study evaluated the toxicity and pharmacokinetics of combining exisulind (150-250 mg) given orally twice daily and docetaxel (30-36 mg/m2) administered intravenously on days 1, 8, and 15 of a 4-week cycle. RESULTS: Twenty patients with a range of advanced solid tumors (median age, 59 years; age range, 35-77 years; median performance status, 1) received a total of 70 courses. Observed adverse events were mild to moderate, and there was no dose-limiting toxicity at any level. Grade 3 gastrointestinal toxicities were present in 10 of the 70 cycles (10%) and included nausea, vomiting, dyspepsia, and elevated alkaline phosphatase. Neutropenia was present in four cycles in patients treated with a docetaxel dose of 36 mg/m2. Pharmacokinetic analysis did not demonstrate a clear effect of exisulind on docetaxel pharmacokinetics and vice versa. Relationships were evident between the plasma concentration of exisulind and the development of grade 2 or greater toxicities. One third of patients maintained stable disease for 3 to 12 cycles, but no objective responses were observed. CONCLUSIONS: The combination of docetaxel (36 mg/m2, weekly) and exisulind (500 mg/d) was reasonably well tolerated, and it is undergoing phase II testing in patients with non-small-cell lung cancer. PMID- 15534097 TI - Plasma coagulation markers in patients with solid tumors and venous thromboembolic disease receiving oral anticoagulation therapy. AB - PURPOSE: To correlate the concentration of plasma coagulation markers at baseline and during follow-up in patients with solid tumors and venous thromboembolic disease with the risk of recurrence and death. EXPERIMENTAL DESIGN: Patients (N = 223) with first episode of venous thromboembolic disease received oral anticoagulation with warfarin for a target international normalized ratio of 2 to 3. Plasma coagulation markers were measured before instituting warfarin and at 3 monthly intervals, thereafter. RESULTS: The median duration of oral anticoagulation was 6.7 months (range 2 weeks to 11 months). Major bleeding episodes occurred in 18 patients (8%), and minor hemorrhagic events occurred in 15 (6.7%) patients. Patients with advanced malignancy (P = 0.032), history of surgery (P = 0.057), and those with poor performance status (P = 0.001) were more likely to encounter major bleeding episodes. Recurrence of venous thromboembolic disease was diagnosed in 31 patients (14%). At univariate analysis, advanced stage of cancer (P = 0.03), performance status > 1 (P = 0.001), treatment with chemotherapy (P = 0.01), the presence of metastatic liver disease (P = 0.03), higher d-dimer (P = 0.001), and thrombin antithrombin complex levels (P = 0.01) were features predictive of recurrent venous thromboembolic disease. At multivariate analysis, poor performance status (P = 0.01) and d-dimer levels (P = 0.001) were predictors of recurrent venous thromboembolic disease. Persistent activation of coagulation as indicated by an upward trend in d-dimer (P = 0.001) and antithrombin (P = 0.001) was observed in patients who developed recurrent thrombosis. Similar upward trends in d-dimer (P = 0.001), antithrombin (P = 0.001), and prothrombin fragment F1 + 2 (P = 0.001) was observed in the 76 patients who died during the study period and in the patients who received chemotherapy. CONCLUSIONS: Successful oral anticoagulation with warfarin in patients with cancer and venous thromboembolic disease is more likely to be achieved in patients with early stage tumors and good performance status. The persistence of activation of hemostasis as shown by plasma coagulation markers is a strong predictor of recurrence and poor outcome. PMID- 15534098 TI - Delivery of a liposomal c-raf-1 antisense oligonucleotide by weekly bolus dosing in patients with advanced solid tumors: a phase I study. AB - PURPOSE: Rapid cleavage in vivo and inefficient cellular uptake limit the clinical utility of antisense oligonucleotides (AON). Liposomal formulation may promote better intratumoral AON delivery and inhibit degradation in vivo. We conducted the first clinical evaluation of this concept using a liposomal AON complementary to the c-raf-1 proto-oncogene (LErafAON). EXPERIMENTAL DESIGN: A dose escalation study was done to determine the maximum tolerated dose and to characterize the toxicities of LErafAON given as weekly intravenous infusion for 8 weeks to adults with advanced solid tumors. Pharmacokinetic analysis and evaluation of c-raf-1 target suppression in peripheral blood mononuclear cells were included. RESULTS: Twenty-two patients received LErafAON (median 7 infusions; range 1-27) at doses of 1, 2, 4, and 6 mg/kg/week. Across all dose cohorts patients experienced infusion-related hypersensitivity reactions including flushing, dyspnea, hypoxia, rigors, back pain, and hypotension. Prolonged infusion duration and pretreatment with acetaminophen, H1- and H2 antagonists, and corticosteroids reduced the frequency and severity of these reactions. Progressive thrombocytopenia was dose-limiting at 6 mg/kg/week. No objective responses were observed. Two patients treated at the maximum tolerated dose of 4 mg/kg/week had evidence of stable disease, with dosing extended beyond 8 weeks. Pharmacokinetic analysis revealed persistence of detectable circulating rafAON at 24 hours in 7 of 10 patients in the highest 2 dose cohorts. Suppression of c-raf-1 mRNA was noted in two of five patients analyzed. CONCLUSIONS: Dose independent hypersensitivity reactions and dose-dependent thrombocytopenia limited tolerance of LErafAON. Future clinical evaluation of this approach will depend on modification of the liposome composition. PMID- 15534099 TI - X-tile: a new bio-informatics tool for biomarker assessment and outcome-based cut point optimization. AB - The ability to parse tumors into subsets based on biomarker expression has many clinical applications; however, there is no global way to visualize the best cut points for creating such divisions. We have developed a graphical method, the X tile plot that illustrates the presence of substantial tumor subpopulations and shows the robustness of the relationship between a biomarker and outcome by construction of a two dimensional projection of every possible subpopulation. We validate X-tile plots by examining the expression of several established prognostic markers (human epidermal growth factor receptor-2, estrogen receptor, p53 expression, patient age, tumor size, and node number) in cohorts of breast cancer patients and show how X-tile plots of each marker predict population subsets rooted in the known biology of their expression. PMID- 15534100 TI - Increased levels of interleukin-10 in serum from patients with hepatocellular carcinoma correlate with profound numerical deficiencies and immature phenotype of circulating dendritic cell subsets. AB - Increased levels of interleukin (IL)-10 have been described as a negative prognostic indicator for survival in patients with various types of cancer. IL-10 exerts tolerogenic and immunosuppressive effects on dendritic cells, which are crucial for the induction of an antitumor immune response. Blood dendritic cell antigen (BDCA)-2 and BDCA-4 are specifically expressed by CD123(bright) CD11c- plasmacytoid dendritic cells; whereas BDCA-1 and BDCA-3 define 2 distinct subsets of CD11c+ myeloid dendritic cells. In this study, the T-helper cell (Th)1/Th2 cytokine serum profile of 65 hepatocellular carcinoma patients was assessed. We found that serum levels of IL-10 were substantially increased in hepatocellular carcinoma patients as compared with controls. Peripheral blood mononuclear cells from healthy volunteers were exposed to recombinant human (rh)IL-10 in vitro to additionally characterize its impact on distinct blood dendritic cell subsets. A dramatic decrease of all myeloid dendritic cell (MDC) and plasmacytoid dendritic cell (PDC) subsets was detectable after 24 hours of continuous rhIL-10 exposure. Moreover, the expression of HLA-DR, CD80 and CD86, was significantly reduced on rhIL-10-treated dendritic cell subsets. Direct ex vivo flow cytometric analysis of various dendritic cell subpopulations in peripheral blood from hepatocellular carcinoma patients revealed an immature phenotype and a substantial reduction of circulating dendritic cells that was associated with increased IL-10 concentrations in serum and with tumor progression. These findings confirm a predominantly immunosuppressive role of IL-10 for circulating dendritic cells in patients with hepatocellular carcinoma and, thus, may indicate novel aspects of tumor immune evasion. PMID- 15534101 TI - Autoantibody profiles and neurological correlations of thymoma. AB - PURPOSE: Determine muscle and neuronal autoantibody frequencies in patients with thymoma, with and without paraneoplastic neurological accompaniments. EXPERIMENTAL DESIGN: Analysis of IgG autoantibodies in stored serum collected between 1985 and 2003 from 201 patients with histologically diagnosed thymoma (including six with thymic carcinoma). Contemporary assays quantitated antibodies reactive with muscle and neuronal cation channels, muscle sarcomeric proteins and neuronal cytoplasmic, and nuclear proteins. RESULTS: Neurological diagnoses included myasthenia gravis (MG), myositis, encephalitis, neuromuscular hyperexcitability, autonomic neuropathy, and subacute hearing loss, a previously unrecognized accompaniment of thymoma. Muscle acetylcholine receptor (AChR) binding antibodies were found in all patients with a diagnosis of MG. Muscle autoantibodies (AChR-binding, AChR-modulating, or striational) were also found in 59% of patients without any neurological disorder. One or more neuronal autoantibodies were found in 41% of patients without any neurological disorder, 43% of patients with MG only, and 78% of patients with other neurological disorders. Neuronal autoantibody specificities were, in descending order of frequency, as follows: glutamic acid decarboxylase, voltage-gated potassium channel, collapsin response-mediator protein-5, ganglionic AChR, and antineuronal nuclear antibody-type 1 (ANNA-1). CONCLUSIONS: Neuronal autoantibodies complement skeletal muscle autoantibodies as serological markers of thymoma in patients with and without clinical evidence of a neurological disorder. The high prevalence of glutamic acid decarboxylase autoantibody, not previously considered a paraneoplastic marker, justifies its consideration as a marker of thymoma-related neurological autoimmunity. Serological evaluation of a patient's profile of neuronal and muscle autoantibodies may aid in preoperative identification of an indeterminate mediastinal mass. PMID- 15534102 TI - Molecular profiling and classification of sporadic renal cell carcinoma by quantitative methylation analysis. AB - PURPOSE: Preoperative histologic classification of solid renal masses remains limited with current technology. We determine the utility of molecular profiling based on quantitative methylation analysis for characterization of sporadic renal cell carcinoma. EXPERIMENTAL DESIGN: Primary renal cell carcinomas representing three different histologic subtypes were obtained from a total of 38 patients who underwent radical nephrectomy for suspected malignant disease. Genomic DNA was isolated from tumors and was subjected to sodium bisulfite modification. The normalized index of methylation (NIM) for each sample was determined by quantitative real-time methylation-specific PCR at 17 different gene promoters. Hierarchical cluster analysis was performed by using an unsupervised neural network with binary tree topology. RESULTS: The majority of gene promoters that were analyzed in this study demonstrated very low levels of methylation (NIM <1.0). The RASSF1A gene promoter, however, was methylated in 30 of 38 (79%) cases. The frequency of RASSF1A methylation in papillary, clear-cell, and oncocytoma subtypes was 100, 90, and 25%, respectively. The highest levels of RASSF1A methylation were observed in the papillary (mean NIM = 78.9) and clear cell (mean NIM = 13.4) subtypes. The vast majority of oncocytomas were completely unmethylated, and none demonstrated >1% methylation (mean NIM = 0.11). Hierarchical cluster analysis based on quantitative methylation levels resulted in stratification of sporadic renal cell carcinomas into their discrete histologic subtypes. CONCLUSIONS: Classification of sporadic renal cell carcinomas into histologic subtypes can be accomplished via multigene quantitative methylation profiling. Validation of this approach and selection of appropriate methylation markers may ultimately lead to use of this technology in the preoperative assessment of suspicious renal masses. PMID- 15534103 TI - Mouse mammary tumor virus-like RNA transcripts and DNA are found in affected cells of human breast cancer. AB - Identifiable risk factors for the development of breast cancer include age, diet, family history, and lifetime estrogen exposure. An infectious agent (mouse mammary tumor virus; MMTV) is known to cause murine breast tumors and may be involved in the pathogenesis of human disease. Multiple studies have detected MMTV-like sequences in 30 to 60% of breast cancer samples and up to 1.8% of samples from normal breast. Using in situ PCR of MMTV-like sequences of formalin fixed, paraffin-embedded breast tissue, viral sequences have been located in cancerous epithelial cells in breast acini of male and female breast tumors, but not in adjacent nonmalignant cells. MMTV-like sequences were also located in the epithelial cells of male gynecomastia samples. Using reverse transcriptase in situ PCR, RNA transcripts from the env gene were also detected within cancerous epithelial cells of 78% of DNA-positive tumors, 80% of gynecomastia samples, and 0% of normal tissues screened. This suggests the virus may be replicating in these cells. The epidemiologic and histopathological data are consistent with the association of an MMTV-like virus with breast cancers in men and women. The association with gynecomastia, a benign, possibly premalignant condition suggests hormonal influences, rather than cancer per se, may be the dominant factor in determining viral presence and replication. PMID- 15534104 TI - Loss of methylthioadenosine phosphorylase and elevated ornithine decarboxylase is common in pancreatic cancer. AB - PURPOSE: Loss of the methylthioadenosine phosphorylase (MTAP) gene at 9p21 is observed frequently in a variety of human cancers. We have shown previously that MTAP can act as a tumor suppressor gene and that its tumor suppressor function is related to its effect on polyamine homeostasis. Ornithine decarboxylase is a key enzyme in the regulation of polyamine metabolism. The aim of this study is to analyze MTAP and ornithine decarboxylase (ODC) expression in primary pancreatic tumor specimens. EXPERIMENTAL DESIGN: We measured MTAP and ODC activity in protein extracts derived from 30 surgically resected tumor samples and eight normal pancreas samples. In a subset of six samples, we also examined MTAP DNA using interphase fluorescence in situ hybridization. In addition, we examined the effect of the ODC inhibitor difluoromethylornithine on two pancreatic adenocarcinoma-derived cell lines. RESULT: MTAP activity was 2.8-fold reduced in adenocarcinomas and 6.3-fold reduced in neuroendocrine tumors compared with control pancreas. Conversely, ODC activity was 3.6-fold elevated in adenocarcinomas and 3.9-fold elevated in neuroendocrine tumors compared with control pancreas. Using interphase fluorescence in situ hybridization, we found in tumor samples that 43 to 75% of the nuclei had lost at least one copy of MTAP locus, indicating that loss of MTAP activity was at least partially because of deletion of the MTAP locus. We also show that inhibition of ODC by difluoromethylornithine caused decreased cell growth and increased apoptosis in two MTAP-deleted pancreatic adenocarcinoma-derived cell lines. CONCLUSIONS: MTAP activity is frequently lost, and ODC activity is frequently elevated in both pancreatic adenocarcinoma and neuroendocrine tumors. Inhibition of ODC activity caused decreased cell growth and increased apoptosis in pancreatic tumor-derived cell lines. These findings suggest that MTAP and polyamine metabolism could be potential therapeutic targets in the treatment of pancreatic cancer. PMID- 15534105 TI - Prognostic value of ERM gene expression in human primary breast cancers. AB - We measured the expression of ERM gene, a nuclear transcription factor belonging to the ets family, in a series of 364 unselected primary breast cancers from patients who underwent locoregional surgery in the Centre Oscar Lambret between May 1989 and December 1991. The expression of ERM was quantified with a real-time one-step reverse transcription-PCR assay based on the 5'-nuclease activity of the TaqDNA polymerase and with an Abi Prism 7700 Sequence Detector System (Applied Biosystems, Courtaboeuf, France). ERM was positively correlated (Spearman test) to epidermal growth factor receptor (EGFR; P < 0.001, r = 0.296) and to histoprognostic grading (P = 0.044, r = 0.112), whereas it was negatively correlated to estradiol receptors (P = 0.019, r = -0.124), HER3 (c-erbB-3; P = 0.01, r = -0.135), and HER4 (c-erbB-4; P = 0.003, r = -0.154). Using the chi2 test, a positive relationship was found between the expression of ERM and EGFR (chi2 = 7.795, P = 0.007). In overall survival studies, Cox univariate analyses demonstrated a prognostic value of ERM (P = 0.006; risk ratio, 2.95) besides the classical prognostic factors histoprognostic grading, node involvement, tumor size, estradiol receptors, progesterone receptors, EGFR, HER3, and HER4. In multivariate analyses, ERM preserved its prognostic value (P = 0.004; risk ratio, 3.779) together with histoprognostic grading, tumor size, estradiol receptors, and progesterone receptors. In relapse-free survival studies, univariate analyses demonstrated that histoprognostic grading, node involvement, tumor size, and HER4 were prognostic factors. These parameters, except histoprognostic grading, retained their prognostic value in multivariate analyses. This study demonstrates for the first time that ERM gene expression is an independent adverse prognostic factor for overall survival in breast cancer patients. PMID- 15534106 TI - Overexpression of Aurora-A contributes to malignant development of human esophageal squamous cell carcinoma. AB - PURPOSE: Aurora-A/STK15/BTAK, a centrosome-associated oncogenic protein, is implicated in the control of mitosis. Overexpression of Aurora-A has been shown to result in chromosomal aberration and genomic instability. Multiple lines of evidence indicate that Aurora-A induces cell malignant transformation. In the current study, we are interested in investigating the expression of Aurora-A in human esophageal squamous cell carcinoma (ESCC) and characterizing the association of Aurora-A with ESCCmalignant progression. EXPERIMENTAL DESIGN: Aurora-A protein expression was examined in 84 ESCC tissues and 81 paired normal adjacent tissues by either immunohistochemistry or Western blot analysis. In addition, a gene-knockdown small interfering RNA technique was used in ESCC cells to investigate whether Aurora-A contributes to the ability of a tumor to grow invasively. RESULTS: The amount of Aurora-A protein in ESCC was considerably higher than that in normal adjacent tissues. Overexpression of Aurora-A was observed in 57 of 84 (67.5%) ESCC samples. In contrast, <2% of normal adjacent tissue displayed high expression of Aurora-A. Interestingly, overexpression of Aurora-A seemed to correlate with the invasive malignancy of ESCC. Disruption of endogenous Aurora-A using small interfering RNA technique substantially suppressed cell migrating ability. CONCLUSION: The findings presented in this report show that Aurora-A expression is elevated in human esophageal squamous cell carcinoma and is possibly associated with tumor invasion, indicating that overexpression of Aurora-A may contribute to ESCC occurrence and progression. PMID- 15534107 TI - Dominant papillary subtype is a significant predictor of the response to gefitinib in adenocarcinoma of the lung. AB - PURPOSE: Gefitinib (IRESSA; AstraZeneca, Osaka, Japan) shows excellent antitumor activity against advanced non-small-cell lung cancer, especially for the treatment of adenocarcinoma. However, the predictive factors for the response to gefitinib are still controversial. The aim of this study was to identify the clinicopathological and immunohistochemical features that are favorable to the use of gefitinib in adenocarcinoma patients. EXPERIMENTAL DESIGN: Between June 2002 and October 2003, 36 adenocarcinoma patients who experienced a relapse after surgical resection were treated with gefitinib at our hospital. The histologic patterns of the tumors were divided into four distinctive subtypes according to the revised World Health Organization histologic classification, and the dominant histologic subtype for the maximum cut surface of each resected specimen was documented. Association between the response to gefitinib and the clinicopathological features or immunohistochemical expression of epidermal growth factor receptor (EGFR), phosphorylated EGFR, or c-erbB-2 were then investigated. RESULTS: A significant association between the response to gefitinib and dominant papillary subtype findings was observed (P = 0.0021); the survival time of papillary subtype patients was also significantly prolonged compared with that of non-papillary subtype patients (P = 0.03). No other clinicopathological features or the expression of EGFR, phosphorylated EGFR, or c erbB-2 were associated with the response to gefitinib. CONCLUSIONS: The results of the present study indicate that dominant papillary subtype findings of lung adenocarcinomas can be an important predictor of the response to gefitinib. Thus, this type of adenocarcinoma might be susceptible to postoperative adjuvant treatment with gefitinib. PMID- 15534108 TI - High expression of PRL-3 promotes cancer cell motility and liver metastasis in human colorectal cancer: a predictive molecular marker of metachronous liver and lung metastases. AB - PURPOSE: Overexpression of PRL-3 has been implicated in colorectal cancer metastases. We investigated the significance of PRL-3 expression in the progression and development of colorectal cancer. EXPERIMENTAL DESIGN: We transfected PRL-3-specific small interfering RNA into human colon cancer DLD-1 cells and analyzed its effect on proliferation, motility, and hepatic colonization. Using an in situ hybridization method, we examined the levels of PRL-3 expression in both primary (177 cases) and metastatic (92 cases) human colorectal cancers and elucidated the relationships with clinicopathological parameters including the incidence of metachronous liver and/or lung metastasis after curative surgery for primary tumor. RESULTS: Transient down-regulation of PRL-3 expression in DLD-1 cells abrogated motility (in vitro) and hepatic colonization (in vivo), but no effect on the proliferation of these cells was observed. In human primary colorectal cancers, the frequency of up-regulated PRL 3 expression in cases with liver (84.4%) or lung (88.9%) metastasis was statistically higher than that in cases without either type of metastasis (liver, 35.9%; lung, 42.3%). In metastatic colorectal cancer lesions, high expression of PRL-3 was frequently detected (liver, 91.3%; lung, 100%). Interestingly, metachronous metastasis was observed more frequently in the cases with high PRL-3 expression (P < 0.0001). CONCLUSIONS: These results indicate that PRL-3 expression in colorectal cancers may contribute to the establishment of liver metastasis, particularly at the step in which cancer cells leave the circulation to extravasate into the liver tissue. In addition, PRL-3 is expected to be a promising biomarker for identifying colorectal cancer patients at high risk for distant metastases. PMID- 15534109 TI - Absence of a correlation between the presence of a single nucleotide polymorphism in the matrix metalloproteinase 1 promoter and outcome in patients of chondrosarcoma. AB - PURPOSE: Increased levels of matrix metalloproteinase 1 (MMP-1) expression have been associated with poor outcome in chondrosarcoma. The existence of a single nucleotide polymorphism creating an Ets-binding site in the MMP-1 promoter may be one mechanism for elevated MMP-1 transcription. The aim of our study was to identify the prevalence of this single nucleotide polymorphism (SNP) in chondrosarcoma patients, to determine its correlation with disease outcome, and to discern whether it could serve as a prognostic marker in patients with chondrosarcoma. EXPERIMENTAL DESIGN: Sixty-seven chondrosarcoma specimens were selected sequentially from an established tumor bank with a median duration of 47 months follow-up (range, 24 to 179 months). DNA was extracted, amplified with PCR, and sequenced to determine presence (GG) or absence of the Ets-binding site created by the SNP. RESULTS: Eighteen (27%) samples were homozygous for the absence of the Ets site, 34 (51%) were heterozygous for the SNP, and 15 (22%) were homozygous for the SNP. The 5-year overall survival rate for patients was 78, 80, and 84%, respectively (P = 0.5527). The disease-free survival rate was 16, 63, and 76%, respectively (P = 0.0801). The 5-year disease-free survival rate for patients with the homozygous G/G genotype was 16%, compared with 71% for patients who were either homozygous or heterozygous for the GG allele (P = 0.0444). CONCLUSIONS: Despite a statistical correlation between MMP-1 gene expression and outcome in chondrosarcoma, this study demonstrates an absence of a correlation between the presence of the SNP and prognosis in patients with chondrosarcoma. PMID- 15534110 TI - Altered expression of metastasis-associated and regulatory molecules in effusions from breast cancer patients: a novel model for tumor progression. AB - PURPOSE: The aim of this study was to characterize phenotypic alterations along the progression of breast carcinoma from primary tumor to pleural effusion through analysis of the expression of proteases, laminin receptors (LRs), and transcription factors involved in invasion and metastasis. EXPERIMENTAL DESIGN: The material studied consisted of 60 malignant pleural effusions from breast cancer patients and 68 corresponding solid tumors (37 primary and 31 metastatic tumors). Expression of matrix metalloproteinases [MMPs (MMP-1, MMP-2, MMP-9, and MMP-14)], the MMP inhibitor tissue inhibitor of metalloproteinase-2, the MMP inducer EMMPRIN, the 67-kDa LR, the alpha6 integrin subunit, and the transcription factors AP-2, Ets-1, and PEA3 was studied using immunohistochemistry, mRNA in situ hybridization, reverse transcription polymerase chain reaction, zymography, and flow cytometry. Hormone receptor (estrogen receptor and progesterone receptor) status and c-erbB-2 status were also studied. RESULTS: Significantly reduced estrogen receptor (P < 0.001) and progesterone receptor (P = 0.001) expression was seen in effusions compared with primary tumors, with opposite findings for c-erbB-2 (P = 0.003). Tumor cell MMP-2 protein expression in effusions was higher than that in primary tumors (P < 0.001) and lymph node metastases (P = 0.01). In situ hybridization demonstrated higher MMP-2 (P = 0.007), PEA3 (P = 0.038), and EMMPRIN (P = 0.026) mRNA expression in effusions. The time to progression from primary tumor to effusion was significantly shorter for patients whose primary tumors expressed MMP-1 (P = 0.016) and who expressed the 67-kDa LR protein in primary tumor (P = 0.007) and effusion (P = 0.015). CONCLUSIONS: Our data provide documented evidence of molecular events that occur during the progression of breast carcinoma from primary tumor to effusion. The coordinated up-regulation of MMP-2 and Ets transcription factors in carcinoma cells in effusions is in full agreement with our previous reports linking these factors to poor prognosis in ovarian cancer. The rapid progression to effusion in cases showing MMP-1 and 67-kDa LR expression in primary tumor cells links aggressive clinical behavior with expression of metastasis-associated molecules in this setting. PMID- 15534111 TI - Mina53 as a potential prognostic factor for esophageal squamous cell carcinoma. AB - PURPOSE: We previously identified mina53, a novel Myc target gene. Here we investigated whether mina53 is related to esophageal squamous cell carcinoma (ESCC), a disease with poor prognosis. EXPERIMENTAL DESIGN: Mina53 expression was suppressed in ESCC cell lines by a RNA interference method to investigate whether Mina53 is involved in cell proliferation. Expression of Mina53 was investigated by Western blotting in tissue sections from patients with ESCC. Immunohistochemical analysis of Mina53 was carried out and compared with that using anti-Ki-67 antibody. Finally, the level of Mina53 expression was compared with the length of survival of patients with ESCC. RESULTS: Reduction of mina53 expression by RNA interference suppressed cell proliferation in ESCC cell lines. Western blot analysis of surgically resected ESCC specimens indicated that the expression of Mina53 in tumors was increased compared with that in adjacent nonneoplastic tissues in all four specimens examined. When formalin-fixed specimens from 52 patients with ESCC were stained immunohistochemically, it was found that Mina53 was highly expressed in 83% of specimens. Anti-Mina53 antibody stained tumors more efficiently than antibody against Ki-67, a cell proliferation biomarker, in some cancer specimens. Patients with high expression of Mina53 had shorter survival periods, whereas the expression level of Ki-67 in ESCC showed no relationship to patient outcome. CONCLUSIONS: Taken together, our results indicate that expression of Mina53 is a characteristic feature of ESCC and suggest that immunostaining by anti-Mina53 antibody may be useful as a potential prognostic indicator. PMID- 15534112 TI - Efficacy of the novel camptothecin gimatecan against orthotopic and metastatic human tumor xenograft models. AB - PURPOSE: Gimatecan, a novel oral lipophilic camptothecin characterized by favorable features at molecular/cellular level and by a promising profile of preclinical activity, is currently in clinical phase I/II. The aim of the study was to additionally investigate the therapeutic potential of the drug in human tumor xenografts growing in different organs as models representative of tumor growth in the clinical setting. EXPERIMENTAL DESIGN: The models include two orthotopic central nervous system tumors, two melanomas growing intracranially, and an ovarian carcinoma growing i.p. In addition, gimatecan was tested against experimental lung metastases of two tumor types (lung and ovarian carcinomas). Gimatecan was delivered by oral gavage according to various schedules (daily or intermittent). The time (in days) mice required to show evident signs of disease was used as end point for drug efficacy. RESULTS: Gimatecan was highly effective in delaying disease manifestations in all tumor systems investigated. In the intracranially growing tumors, a significant time increase (versus control mice) was achieved by the drug administered according to all of the schedules. In addition, almost all treated mice were alive and tumor-free at the end of the experiment in the metastatic models and in the ascitic ovarian tumor. The daily prolonged treatment schedule was the best one. CONCLUSIONS: In all tumor systems investigated, including orthotopic tumor growth models and lung metastases, the oral administration of gimatecan showed a therapeutic benefit in terms of survival increase. The good oral availability allowed a prolonged daily treatment regimen, which seems the most promising to exploit the therapeutic potential of the drug. PMID- 15534113 TI - Potent antitumor activity of novel iron chelators derived from di-2-pyridylketone isonicotinoyl hydrazone involves fenton-derived free radical generation. AB - PURPOSE: The development of novel and potent iron chelators as clinically useful antitumor agents is an area of active interest. Antiproliferative activity of chelators often relates to iron deprivation or stimulation of iron-dependent free radical damage. Recently, we showed that novel iron chelators of the di-2 pyridylketone isonicotinoyl hydrazone (PKIH) class have potent and selective antineoplastic activity (E. Becker, et al., Br. J. Pharmacol., 138: 819-30, 2003). In this study, we assessed the effects of the PKIH analogues on the redox activity of iron in terms of understanding their antitumor activity. EXPERIMENTAL DESIGN: We tested the PKIH analogues for their ability to promote iron-mediated ascorbate oxidation, benzoate hydroxylation, and plasmid degradation. Subsequent experiments assessed their ability to bind DNA, inhibit topoisomerase I, and cause DNA damage. To measure intracellular reactive oxygen species, we used the redox-sensitive probe, 2',7'-dichloro-fluorescein-diacetate, to measure intracellular PKIH-dependent redox activity. RESULTS: The PKIH analogues had relatively little effect on ascorbate oxidation in the presence of Fe(III) but stimulated benzoate hydroxylation and plasmid DNA degradation in the presence of Fe(II) and H2O2. These ligands could not inhibit DNA topoisomerase I or cause DNA damage in intact cells. PKIH markedly increased the intracellular generation of reactive oxygen species, and this was inhibited by catalase. This enzyme also decreased the antiproliferative effect of PKIH, indicating H2O2 played a role in its cytotoxic activity. CONCLUSIONS: Our results suggest that the antiproliferative effects of these chelators relates to intracellular iron chelation, followed by the stimulation of iron-mediated free radical generation via the so-formed iron complex. PMID- 15534114 TI - Effects of exemestane and tamoxifen in a postmenopausal breast cancer model. AB - PURPOSE: To optimize treatment strategies for postmenopausal breast cancer patients, we investigated the efficacy of the steroidal aromatase inhibitor exemestane alone or in combination with the antiestrogen tamoxifen in a xenograft model of postmenopausal breast cancer. We also determined the effects of these agents in sequential second-line therapy and the effect of the nonsteroidal aromatase inhibitor letrozole on tumors that progressed on the above treatments. EXPERIMENTAL: Aromatase-transfected human estrogen receptor-positive breast cancer cells (MCF-7Ca) were grown as tumors in ovariectomized athymic mice. Animals received subcutaneous injection with vehicle, tamoxifen, exemestane, tamoxifen plus exemestane, and letrozole. Tumor volumes were measured weekly. RESULTS: All treatments were effective initially in suppressing tumor growth as first-line therapy compared with vehicle treatment. Exemestane suppressed tumor growth to a greater extent than tamoxifen. However, the combination of tamoxifen plus exemestane was more effective than either drug alone. After tumor volumes doubled on initial treatment, the mice were crossed over to receive exemestane or tamoxifen. Tumor growth slowed briefly in mice treated with tamoxifen and crossed over to exemestane, but tumor growth continued unabated in those changed from exemestane to tamoxifen. However, letrozole was effective in both groups as third line therapy for a limited period. Letrozole as initial single agent was the best overall treatment in terms of the degree of tumor suppression and the length of effectiveness of treatment. CONCLUSION: Exemestane was more effective in controlling tumor growth than tamoxifen. In addition, the combination of exemestane plus tamoxifen was clearly more effective than sequential use of these agents in the tumor model. However, the nonsteroidal aromatase inhibitor letrozole as first-line therapy was overall the most effective treatment in controlling tumor growth. PMID- 15534115 TI - Antitumor activity of hydrophilic Paclitaxel copolymer prodrug using locoregional delivery in human orthotopic non-small cell lung cancer xenograft models. AB - Paclitaxel (Taxol) has demonstrated clinical activity in non-small-cell lung cancer (NSCLC), but its use has not led to marked improvements in survival. This ineffectiveness can in part be attributed to inadequate delivery of effective drug levels to the lung via systemic administration and to drug resistance mechanisms. Locoregional drug administration and the use of drug copolymers are possible approaches to address these issues. In this study, we evaluated the activity of a poly(L-glutamic acid)-paclitaxel (PGA-TXL) formulation administered by intratracheal injection to mice bearing orthotopic human NSCLC tumors (H460, H358). H460 cells were found to be sensitive to paclitaxel and PGA-TXL in vitro, in a time- and concentration-dependent manner. In preliminary acute toxicity studies, PGA-TXL administered by intratracheal injection was found to be much less toxic than paclitaxel, as anticipated. Mice into which H460 cells had been implanted by intratracheal injection were given single-dose intratracheal treatments with paclitaxel (1.2 or 2.4 mg/kg) or with PGA-TXL (15 mg/kg, paclitaxel equivalents) 1 week later. When the mice were sacrificed at up to 65 days after tumor implantation, they were evaluated grossly for tumor at bronchial, neck, and lung sites. Control mice had tumors in 60% of all three sites, and all of the control mice had tumors in at least one site. The low- and high-dose Taxol groups had fewer incidences at these three sites (27-33%) and 60 80% of these mice had tumors in at least one site. The PGA-TXL mice displayed a low (13%) incidence at these sites, and only 40% had detectable tumors. In a subsequent survival study with the intratracheal H358 model, control mice had a mean life span of 95 days, whereas both the intratracheal Taxol (2.5 mg/kg, every 7th day for three doses) and the intratracheal PGA-TXL (20 mg/kg, paclitaxel equivalents, every 7th day for three doses) groups had improved survival (mean life spans: 133.5 and 136.5 days, respectively). In pilot studies intended to compare the feasibility of the development of paclitaxel aerosols suitable for clinical application, based either on Cremophor solutions or on PGA backbones, only the latter gave acceptable particle size distributions and flow rates. These results encourage the development and application of Cremophor-free copolymer formulations of paclitaxel for locoregional treatment (e.g., as aerosol) of endobronchial malignant diseases. PMID- 15534116 TI - Androgen receptor is targeted to distinct subcellular compartments in response to different therapeutic antiandrogens. AB - PURPOSE: Antiandrogens are routinely used in the treatment of prostate cancer. Although they are known to prevent activation of the androgen receptor (AR), little is known about the mechanisms involved. This report represents the first study of the localization of wild-type AR following expression at physiologic relevant levels in prostate cells and treatment with androgen and antiandrogens. EXPERIMENTAL DESIGN: We have characterized a cellular model for prostate cancer using in situ cellular fractionation, proteomics, and confocal microscopy and investigated the effect of antiandrogens in clinical use on the subcellular localization of the AR. RESULTS: Different antiandrogens have diverse effects on the subcellular localization of the AR. Treatment with androgen results in translocation from the cytoplasm to the nucleoplasm, whereas the antiandrogens hydroxyflutamide and bicalutamide lead to reversible association with the nuclear matrix. In contrast, treatment with the antiandrogen cyproterone acetate results in AR association with cytoplasmic membranes and irreversible retention within the cytoplasm. In addition, we demonstrate that AR translocation requires ATP and the cytoskeleton, regardless of ligand. CONCLUSIONS: These results reveal that not all antiandrogens work via the same mechanism and suggest that an informed sequential treatment regime may benefit prostate cancer patients. The observed subnuclear and subcytoplasmic associations of the AR suggest new areas of study to investigate the role of the AR in the response and resistance of prostate cancer to antiandrogen therapy. PMID- 15534117 TI - Myeloma cells are highly sensitive to the granule exocytosis pathway mediated by WT1-specific cytotoxic T lymphocytes. AB - PURPOSE: Because WT1 is a universal tumor antigen, we examined the sensitivity of myeloma cells to WT1-specific cytotoxic T lymphocyte (CTL)-mediated cytotoxicity. EXPERIMENTAL DESIGN: WT1 expression in hematologic malignant cells was examined by quantitative reverse transcription-polymerase chain reaction. The cytotoxicity of a WT1-specific CTL clone against hematologic malignant cells, including myeloma cells, was examined by standard chromium-51 release assays. The extent of membrane damage induced by purified perforin was examined. Induction of WT1 specific CTLs from the patients with multiple myeloma (MM) was attempted, and we examined their function against myeloma cells. RESULTS: The expression levels of WT1 mRNA in myeloma and lymphoma cells were significantly lower than that in acute leukemia cells. Although the WT1 expression levels in myeloma and lymphoma cells were almost same, only myeloma cells were lysed efficiently by WT1-specific CTLs in a HLA-restricted manner. The amounts of interferon-gamma produced by WT1 specific CTLs in response to stimulation with myeloma cells and with lymphoma cells were almost the same, suggesting that WT1 protein is processed and expressed in the context of HLA class I molecules similarly on both myeloma and lymphoma cells. The extent of membrane damage induced by purified perforin appeared to be significantly higher in myeloma cells than in lymphoma cells. WT1 specific CTLs appeared to be present in patients with MM. CONCLUSIONS: The present study has shown that susceptibility of membranes to perforin is an important factor determining the sensitivity of target cells to CTL-mediated cytotoxicity and that WT1 is an ideal target antigen for cellular immunotherapy of MM. PMID- 15534118 TI - Therapeutic synergy of oral taxane BMS-275183 and cetuximab versus human tumor xenografts. AB - PURPOSE: Combination therapy consisting of an oral taxane, BMS-275183, and the anti-epidermal growth factor receptor monoclonal antibody, cetuximab, was assessed for enhanced therapeutic benefit in preclinical tumor models. EXPERIMENTAL DESIGN: Mice bearing human tumor xenografts, either L2987 lung or GEO colon carcinoma, were administered the aforementioned treatments singly or in combination regimens. Delays in tumor growth and tumor-free status were evaluated and combination treatments were assessed relative to optimal solo treatments. RESULTS: Combination therapies with the oral taxane plus cetuximab were tolerated and therapeutic synergistic outcomes obtained. The therapeutic enhancements were >1 log cell kill greater than the antitumor effect caused by either solo agent applied optimally. For example, at the maximum-tolerated dose of BMS-275183, 60 mg/kg/administration, given p.o. once every 3 days for a total of six administrations (q3dx6), 1.0 gross log cell kill was achieved in mice bearing well established (100 to 200 mg) s.c. implanted L2987 tumors. Cetuximab, at an optimal dose of 1 mg/mouse, given i.p. q3dx6, produced 1.3 log cell kill. When cetuximab, 1 mg/mouse, i.p., plus BMS-275183, 25 mg/kg/administration, p.o., were both given q3dx6, the result was 2.6 log cell kill with three of eight mice cured (P < 0.01). Similar efficacy benefits were obtained in the GEO tumor model. CONCLUSIONS: The combination of oral taxane BMS-275183 plus cetuximab provided therapeutically synergistic antitumor activity in two different human tumor xenograft models. Clinical evaluation of this combination is recommended. PMID- 15534119 TI - Antiproliferative effects of interferon-alphaCon1 on ovarian clear cell adenocarcinoma in vitro and in vivo. AB - PURPOSE: We examined the antiproliferative effect of IFN-alphaCon1 and its mechanism on ovarian clear cell adenocarcinoma in vitro and in vivo. EXPERIMENTAL DESIGN: (a) The effects of IFN-alphaCon1 on growth, morphology, cell cycle, and type I IFN-alpha receptor (IFNAR-2) expression were examined on two ovarian clear cell adenocarcinoma cell lines (KOC-5C and KOC-7C) in vitro. (b) KOC-5C or KOC-7C cells were transplanted into nude mice, and changes in tumor volume, tumor weight, apoptosis, necrosis, and microvessel density were investigated. The expression of angiogenesis factors was examined in the serum and the developed tumors. RESULTS: Both cell lines expressed IFNAR-2 mRNA, but its protein was detected only in KOC-7C. In KOC-7C cells, antiproliferative effects were observed in a time- and dose-dependent manner and cell division was blocked at the S phase. The KOC-7C tumors showed decreases in tumor volume and weight; a decreasing tendency in basic fibroblast growth factor (bFGF), vascular endothelial growth factor, and interleukin (IL)-8 protein expression in the tumor; a significant decrease in bFGF and IL-8 protein expression in the serum, and of microvessel density; and significant increase in apoptosis and necrosis in the tumor. In the KOC-5C tumors, these in vitro and in vivo changes were not apparent, and the antiproliferative effects of IFN-alphaCon1 were not obvious. CONCLUSIONS: IFN-alphaCon1 suppresses tumor proliferation by inducing apoptosis, blocking the cell cycle, and inhibiting tumor angiogenesis. Our findings show that the clinical efficacy of IFN-alphaCon1 can be predicted by examining IFNAR-2 expression on tumor cells, and the efficacy of IFN-alphaCon1 treatment can be evaluated by measuring serum bFGF and IL-8 levels. PMID- 15534120 TI - Type I collagen promotes the malignant phenotype of pancreatic ductal adenocarcinoma. AB - PURPOSE: The purpose of this study was to determine the role of functional interactions between pancreatic cancer cells and pancreatic stellate cells (PSCs) in the formation of the desmoplastic reaction (DR) in pancreatic cancer and to characterize the effect of type I collagen (the predominant component of the DR) on pancreatic cancer cell phenotype. EXPERIMENTAL DESIGN: PSCs and type I collagen were identified in sections of pancreatic cancer using immunohistochemistry, and their anatomic relationship was studied. Interactions among pancreatic cancer cell lines (MIA PaCa-2, Panc-1, and AsPC-1), primary cultures of human PSCs, and type I collagen were investigated in a series of tissue culture models. RESULTS: In vivo, the DR causes gross distortion of normal pancreas, bringing cancer cells into close contact with numerous PSCs and abundant type I collagen. In tissue culture models of pancreatic cancer, conditioned media from each cell line increased PSC [3H]thymidine incorporation up to 6.3-fold that of controls, and AsPC-1 cells also increased PSC collagen synthesis 1.3-fold. Type I collagen was observed to increase long-term survival of pancreatic cancer cells treated with 5-fluorouracil, by up to 62% in clonogenic assays. This was because type I collagen increased the proliferation of cancer cells ([3H]thymidine incorporation was up to 2.8-fold that of cells cultured on tissue culture plastic) and reduced apoptosis of AsPC-1 cells in response to 5-fluorouracil (by regulating mcl-1). CONCLUSIONS: These experiments elucidate a mechanism by which the DR in pancreatic cancer may form and, via the collagen within it, promote the malignant phenotype of pancreatic cancer cells, suggesting significant detriment to the host. PMID- 15534121 TI - Surgical removal vs observation for subfoveal choroidal neovascularization, either associated with the ocular histoplasmosis syndrome or idiopathic: I. Ophthalmic findings from a randomized clinical trial: Submacular Surgery Trials (SST) Group H Trial: SST Report No. 9. AB - OBJECTIVE: To present visual acuity findings and related outcomes from eyes of patients enrolled in a randomized trial conducted by the Submacular Surgery Trials (SST) Research Group (SST Group H Trial) to compare surgical removal vs observation of subfoveal choroidal neovascular lesions that were either idiopathic or associated with ocular histoplasmosis. METHODS: Eligible patients 18 years or older had subfoveal choroidal neovascularization (new or recurrent) that included a classic component on fluorescein angiography and best-corrected visual acuity of 20/50 to 20/800 in 1 eye ("study eye"). Patients were examined 3, 6, 12, and 24 months after enrollment to assess study outcomes and adverse events. Best-corrected visual acuity was measured by a masked examiner at the 24 month examination. A successful outcome was defined a priori as 24-month visual acuity better or no more than 1 line (7 letters) worse than at baseline. RESULTS: Among 225 patients enrolled (median visual acuity 20/100), 113 study eyes were assigned to observation and 112 to surgery. Forty-six percent of the eyes in the observation arm and 55% in the surgery arm had a successful outcome (success ratio, 1.18; 95% confidence interval, 0.89-1.56). Median visual acuity at the 24 month examination was 20/250 among eyes in the observation arm and 20/160 for eyes in the surgery arm. The prespecified subgroup of eyes with visual acuity worse than 20/100 at baseline (n = 92) had more successes with surgery; 31 (76%) of 41 eyes in the surgery arm vs 20 (50%) of 40 eyes in the observation arm examined at 24 months (success ratio, 1.53; 95% confidence interval, 1.08-2.16). Five (4%) of 111 eyes in the surgery arm subsequently had a rhegmatogenous retinal detachment. Twenty-seven (24%) of 112 initially phakic eyes in the surgery arm (none in the observation arm) had cataract surgery during follow-up, all among patients older than 50 years. Recurrent choroidal neovascularization developed by the 24-month examination in 58% of surgically treated eyes. CONCLUSIONS: Overall, findings supported no benefit or a smaller benefit to surgery than the trial was designed to detect. Findings support consideration of surgery for eyes with subfoveal choroidal neovascularization and best-corrected visual acuity worse than 20/100 that meet other eligibility criteria for the SST Group H Trial. Other factors that may influence the treatment decision include the risks of retinal detachment, cataract among older patients, and recurrent choroidal neovascularization and the possibility that additional treatment will be required after submacular surgery. PMID- 15534122 TI - Surgical removal vs observation for subfoveal choroidal neovascularization, either associated with the ocular histoplasmosis syndrome or idiopathic: II. Quality-of-life findings from a randomized clinical trial: SST Group H Trial: SST Report No. 10. AB - OBJECTIVE: To report findings regarding health-related quality-of-life outcomes by treatment arm, both overall and within subgroups defined by selected baseline characteristics, among patients who participated in the Submacular Surgery Trials (SST) randomized trial of observation vs surgical removal of subfoveal choroidal neovascular lesions that were either idiopathic or associated with ocular histoplasmosis (SST Group H Trial). DESIGN AND METHODS: Eligible patients were 18 years or older and had subfoveal choroidal neovascularization (including a classic component on fluorescein angiography) and visual acuity of 20/50 to 20/800 inclusive in the eye randomly assigned to surgery or observation. Trained interviewers, who were masked to treatment assignment, administered the National Eye Institute Visual Function Questionnaire (NEI-VFQ), the 36-Item Short-Form Health Survey (SF-36), and the Hospital Anxiety and Depression Scale (HADS) by telephone before enrollment and at 6, 12, and 24 months after enrollment; early enrollees also had interviews at 36 and 48 months. Baseline clinical examinations provided data regarding visual acuity, other aspects of vision, and presence of unilateral or bilateral choroidal neovascularization. RESULTS: Of 225 patients interviewed at baseline and enrolled, 201, 190, and 161 patients (respectively, 89%, 85%, and 88% of those eligible to be interviewed) were interviewed 12, 24, and 36 months later. The NEI-VFQ scores in both treatment arms improved from baseline (median score, 75) to the 24-month interview. Patients in the surgery arm had 4-point larger improvements, on average, than patients in the observation arm (95% confidence interval, 1-8 points). The largest differences between treatment arms for mean 24-month improvements from baseline were in the role difficulties subscale (9 points) and dependency subscale (8 points), with larger mean improvements in the surgery arm. Scores on the SF-36 worsened by approximately 1 point on the physical component summary and improved by 2 points or more on the mental component summary in both treatment arms by 24 months in comparison to baseline. The percentage of HADS-defined "definite cases" of both anxiety and depression declined from baseline in each treatment arm. CONCLUSIONS: Vision-targeted quality of life improved more after submacular surgery than with observation, supporting a possible small overall benefit of surgery suggested by the ophthalmic outcomes reported elsewhere. Ophthalmologists and patients similar to those who participated in the SST Group H Trial should be aware of the effects of submacular surgery on specific aspects of vision-targeted quality of life as well as on ophthalmic outcomes when considering this treatment approach. PMID- 15534123 TI - Risks of progression of retinopathy and vision loss related to tight blood pressure control in type 2 diabetes mellitus: UKPDS 69. AB - OBJECTIVE: To determine the relationship between tight blood pressure (BP) control and the different aspects of diabetic retinopathy in patients with type 2 diabetes mellitus (DM). SETTING: Nineteen hospital-based clinics in England, Scotland, and Northern Ireland. DESIGN: Outcome of retinopathy status assessed by 4-field retinal photography related to allocation within a randomized controlled trial comparing a tight BP control policy aiming for a BP less than 150/85 mm Hg with a less tight BP control policy aiming for a BP less than 180/105 mm Hg. SUBJECTS: One thousand one hundred forty-eight hypertensive patients with type 2 DM were studied. These patients had type 2 DM for a mean duration of 2.6 years at the inception of the Hypertension in Diabetes Study, had a mean age of 56 years; and had a mean BP of 160/94 mm Hg. Seven hundred fifty-eight patients were allocated to a tight BP control policy with angiotensin-converting enzyme inhibitor or beta-blockers as the main therapy; 390 were allocated to a less tight BP control policy. MAIN OUTCOME MEASURES: Deterioration of retinopathy (>/=2-step change on a modified Early Treatment Diabetic Retinopathy Study [ETDRS] final scale), together with end points (photocoagulation, vitreous hemorrhage, and cataract extraction) and analysis of specific lesions (microaneurysms, hard exudates, and cotton-wool spots). Visual acuity was assessed at 3-year intervals using ETDRS logarithm of the minimum angle of resolution charts. Blindness was monitored as an end point with the criterion of Snellen chart assessment at 6/60 or worse. RESULTS: By 4.5 years after randomization, there was a highly significant difference in microaneurysm count with 23.3% in the tight BP control group and 33.5% in the less tight BP control group having 5 or more microaneurysms (relative risk [RR], 0.70; P = .003). The effect continued to 7.5 years (RR, 0.66; P<.001). Hard exudates increased from a prevalence of 11.2% to 18.3% at 7.5 years after randomization with fewer lesions found in the tight BP control group (RR, 0.53; P<.001). Cotton-wool spots increased in both groups but less so in the tight BP control group which had fewer cotton-wool spots at 7.5 years (RR, 0.53; P<.001). A 2-step or more deterioration on the ETDRS scale was significantly different at 4.5 years with fewer people in the tight BP control group progressing 2 steps or more (RR, 0.75; P = .02). Patients allocated to tight BP control were less likely to undergo photocoagulation (RR, 0.65; P = .03). This difference was driven by a difference in photocoagulation due to maculopathy (RR, 0.58; P = .02). The cumulative incidence of the end point of blindness (Snellen visual acuity, >/=6/60) in 1 eye was 18/758 for the tight BP control group compared with 12/390 for less tight BP control group. These equate to absolute risks of 3.1 to 4.1 per 1000 patient years, respectively (P = .046; RR, 0.76; 99% confidence interval, 0.29-1.99). There was no detectable difference in outcome between the 2 randomized therapies of angiotensin-converting enzyme inhibition and beta-blockade. CONCLUSIONS: High BP is detrimental to each aspect of diabetic retinopathy; a tight BP control policy reduces the risk of clinical complications from diabetic eye disease. PMID- 15534124 TI - Detection of age-related macular degeneration using a nonmydriatic digital camera and a standard film fundus camera. AB - OBJECTIVE: To compare gradings of lesions associated with age-related macular degeneration (AMD) from digital and stereoscopic film images. DESIGN: Instrument validation study. PARTICIPANTS: Sixty-two subjects (124 eyes) with varying degrees of AMD, including no AMD. METHODS: Images of the optic disc and macula were taken using a 45 degrees digital camera (6.3 megapixels) through dark adapted pupils and pharmacologically dilated pupils. In addition, 30 degrees stereoscopic retinal film images were taken through pharmacologically dilated pupils of the same eyes. All images were graded for drusen size, type, and area; pigmentary abnormalities; geographic atrophy; and neovascular lesions using the modified Wisconsin Age-Related Maculopathy Grading System. Exact agreement and unweighted kappa scores were calculated for paired gradings resulting from digital and film images. MAIN OUTCOME MEASURE: Agreement between gradings obtained from stereoscopic slide transparencies and digital nonstereoscopic images. RESULTS: Exact agreement between gradings of digital and stereoscopic film images taken through pharmacologically dilated pupils was 91% (kappa = 0.85) for the categories of none, early AMD, and late AMD. Exact agreement for gradings of digital images taken through dark-adapted pupils compared with gradings of film images was 80% (kappa = 0.69). Exact agreement for gradings of digital images captured through dark-adapted and pharmacologically dilated pupils was 86% (kappa = 0.78). In addition, kappa scores for agreement between different approaches for individual lesions were moderate to almost perfect. CONCLUSIONS: Gradings resulting from high-resolution digital images, especially when the pupil is pharmacologically dilated, are comparable with those resulting from film-based images. We conclude that digital imaging of the retina is useful for epidemiological studies of AMD. PMID- 15534126 TI - Palladium 103 (103Pd) plaque radiation therapy for circumscribed choroidal hemangioma with retinal detachment. AB - OBJECTIVE: To describe clinical experience with palladium 103 ((103)Pd) ophthalmic plaque radiotherapy for choroidal hemangioma. METHODS: One course of (103)Pd ophthalmic plaque radiotherapy was used in each of 5 patients with circumscribed choroidal hemangioma who had progressive loss of vision due to subretinal exudation. A mean apex dose of 2900 cGy (2900 rad) was delivered. Functional tests of outcome included best-corrected visual acuity. Anatomic results included changes in tumor height and subretinal fluid documented by ophthalmoscopy, fluorescein angiography, and ultrasonography. RESULTS: All patients had complete resolution of subretinal fluid with reattachment of the retina. All tumors decreased in height (mean, 50%) after treatment. Three patients (60%) demonstrated improvement in visual acuity at the last follow-up, and in 1 patient vision remained stable with resolution of metamorphopsia. Twenty four months after treatment, 1 patient whose visual acuity had recovered from 20/160 to 20/32 had a loss of vision to 20/160 because of radiation maculopathy. For all patients, a mean visual acuity improvement of 2 lines was documented (95% confidence interval, 0.23-0.88). Mean follow-up was 18.6 months (range, 6-29 months). CONCLUSIONS: A single (103)Pd plaque radiation treatment was effective in decreasing tumor height, eliminating subretinal fluid, and improving visual acuity in patients with symptomatic circumscribed choroidal hemangiomas. PMID- 15534125 TI - Scanning laser entoptic perimetry for the detection of age-related macular degeneration. AB - OBJECTIVE: To determine the sensitivity and specificity of scanning laser entoptic perimetry for detecting visual function damage due to age-related macular degeneration (ARMD). METHODS: We measured the presence or absence of visual field disturbances by entoptic perimetry and determined the severity of ARMD based on masked readings of fundus photographs. A prospective masked study comparing the findings of entoptic perimetry with fundus photographs was performed. We recruited 91 patients with ARMD and 24 patients without ARMD during ophthalmologic visits. An appropriate institutional review board approval was obtained for the project. The main outcome measure was the detection of visual scotomata. RESULTS: Scanning laser entoptic perimetry had an overall sensitivity of 82% and a specificity of 100% for the detection of ARMD. The sensitivity for early stages of the disease is greater than 70%, and increases to above 90% for moderate to late stages. CONCLUSION: Scanning laser entoptic perimetry is a specific and sensitive test for detecting ARMD, even at the earliest stages when patients are typically asymptomatic. PMID- 15534127 TI - Health-related quality of life at age 10 years in very low-birth-weight children with and without threshold retinopathy of prematurity. AB - OBJECTIVE: To describe parental perspectives on health status and health-related quality of life (HRQL) at age 10 years in children with birth weights less than 1251 g who participated in the multicenter Cryotherapy for Retinopathy of Prematurity (CRYO-ROP) study. METHODS: In 244 participants in the randomized CRYO ROP trial and 102 CRYO-ROP participants who did not develop ROP, the Health Utilities Index (HUI) system was used to characterize health status for the following 8 attributes: vision, hearing, speech, ambulation, dexterity, emotion, cognition, and pain. Using a utility formula, HRQL was determined for each child on a scale from 0.0 (dead) to 1.00 (perfect health). RESULTS: The proportion of the ROP-randomized group with limitations in 4 attributes or more was 20.6% compared with 2.0% for the no-ROP group. Within the ROP-randomized group, the proportion of "sighted" children with limitations in 4 attributes or more was 6.4% vs 46.5% in the "blind/low vision" group. The median HRQL score for the ROP randomized children was lower than for the no-ROP children (0.72 vs 0.97, P<.001); the median HRQL score for the sighted-randomized children was 0.87 vs 0.27 for the blind/low vision children (P<.001). CONCLUSIONS: Threshold ROP was associated with functional limitations in health attributes and reduction in HRQL scores at age 10 years. Furthermore, among children who developed threshold ROP, a greater reduction in HRQL scores was found among children with a poor visual outcome compared with those with better sight. PMID- 15534128 TI - Safety and efficacy of 2% pirenzepine ophthalmic gel in children with myopia: a 1 year, multicenter, double-masked, placebo-controlled parallel study. AB - OBJECTIVE: To evaluate the safety and efficacy of the relatively selective M(1) antagonist pirenzepine hydrochloride in slowing the progression of myopia in school-aged children. METHODS: This was a parallel-group, placebo-controlled, double-masked study in healthy children, aged 8 to 12 years, with a spherical equivalent of -0.75 to -4.00 diopters (D) and astigmatism of 1.00 D or less. Patients underwent a baseline complete eye examination and regular examinations during a 1-year period. The setting was 13 US academic clinics and private practices. Patients were randomized in a 2:1 ratio to receive 2% pirenzepine ophthalmic gel or a placebo control twice daily for 1 year. RESULTS: At study entry, the spherical equivalent was mean +/- SD -2.098 +/- 0.903 D for the pirenzepine group (n = 117) and -1.933 +/- 0.825 D for the placebo group (n = 57, P = .22). At 1 year, there was a mean increase in myopia of 0.26 D in the pirenzepine group vs 0.53 D in the placebo group (P < .001). No patients in the placebo group and 13 (11%) of 117 patients in the pirenzepine group discontinued participation in the study because of adverse effects (5 [4%] of 117 due to excessive antimuscarinic effects). CONCLUSIONS: Pirenzepine is effective and relatively safe in slowing the progression of myopia during a 1-year treatment period. PMID- 15534129 TI - In vivo confocal microscopy in patients with central cloudy dystrophy of Francois. AB - OBJECTIVE: To report in vivo corneal confocal microscopic findings of patients with central cloudy dystrophy of Francois. METHODS: Two unrelated patients, a 78 year-old man and a 75-year-old woman, with central cloudy dystrophy of Francois were examined using routine slitlamp biomicroscopy and confocal microscopy. RESULTS: In both cases, slitlamp biomicroscopy showed bilateral polygonal opacities separated by clear spaces. The corneal opacities were most prominent centrally and were located in the deeper stromal layer immediately anterior to the Descemet membrane. By confocal microscopy, normal superficial and basal epithelial layers, midstromal layers, and endothelial layers were noted in both cases. However, small highly refractile granules and deposits were observed in the anterior stromal layer in both cases. Also, multiple dark striae among the extracellular matrix with increased intensities were observed in the posterior stroma adjacent to the corneal endothelial layer in both cases. CONCLUSIONS: Abnormal stromal deposits and multiple dark striae were observed in central cloudy dystrophy of Francois using in vivo corneal confocal microscopy. Use of confocal microscopy to investigate these abnormal stromal opacities may be helpful in differentiating various corneal stromal pathologic features. PMID- 15534130 TI - A study of iridectomy histopathologic features of latanoprost- and non latanoprost-treated patients. AB - OBJECTIVES: To examine the histopathologic features of iridectomy specimens from patients undergoing glaucoma surgery and to compare histologic abnormalities in a group of patients with a history of latanoprost therapy with those in a group of patients who had no history of prostaglandin therapy (controls). METHODS: Iridectomy specimens and patient history forms were submitted to the central Latanoprost Pathology Center. These were independently examined by 3 ophthalmic pathologists in a masked fashion. Specimens were evaluated for malignant, premalignant, and other changes including differences in levels of pigmentation, degrees of cellularity, inflammation, vascular abnormalities, and changes in the iris pigment epithelium. RESULTS: Specimens were received from 449 patients with a history of latanoprost treatment and 142 patients who had no history of treatment with latanoprost or other prostaglandin analogues. No evidence of malignant or premalignant changes was found. In latanoprost-treated irides, the prevalence of iris freckles was higher (P = .001) than in control irides, as was the combined number of stromal fibroblasts and melanocytes (P<.001). In a subgroup of specimens received through June 2002, there was no significant difference in mean melanocyte counts (P=.35) obtained by immunohistochemical staining techniques between the latanoprost-treated and control groups. CONCLUSIONS: These findings support previous studies indicating that latanoprost induced eye color changes are due to an increased amount of melanin within the iris stromal melanocytes. The increased numbers of freckles may be a focal manifestation of this effect. PMID- 15534132 TI - 193-nm excimer laser-induced fluorescence detection of fluoroquinolones in rabbit corneas. AB - OBJECTIVE: To measure the 193-nm excimer laser-induced fluorescence of fluoroquinolone-treated cadaver rabbit corneas. METHODS: Prior to ablation with a commercially available ophthalmic excimer laser (Nidek EC-5000; Nidek Technologies, Pasadena, Calif), 35 cadaver rabbit corneas were treated with topical sterile balanced salt solution, 0.3% tobramycin sulfate, or the fluoroquinolones-0.3% ofloxacin, 0.5% levofloxacin, 0.3% ciprofloxacin hydrochloride, or 0.3% gatifloxacin. The fluorescence generated from each ablated corneal layer was measured and used to identify the presence of antibiotic. This was achieved by training a partial least-squares model to discriminate between the fluorescence spectra of antibiotic-treated and antibiotic-free (healthy) cornea. Antibiotic concentrations down to 0.06 microg/mL were detected with high accuracy. Assuming a constant ablation rate of 0.3 microm per laser pulse, the number of corneal layers ablated to reach antibiotic-free cornea is used to calculate the penetration depth of the antibiotic. RESULTS: The mean +/- SD penetration to a detectable depth was as follows: 0.3% ofloxacin, 7.1 +/- 3.0 microm; 0.5% levofloxacin, 6.7 +/- 1.4 microm; 0.3% ciprofloxacin, 1.2 +/- 0.6 microm; and 0.3% gatifloxacin, 7.0 +/- 1.9 microm. The penetration depth of 0.3% tobramycin could not be determined because its fluorescence spectrum overlapped with that of the native cornea. CONCLUSIONS: Topical administration of fluoroquinolone-containing solutions results in measurable differences in laser induced corneal fluorescence. Under these experimental conditions, 0.3% ofloxacin, 0.5% levofloxacin, and 0.3% gatifloxacin all appear to penetrate the epithelium significantly more than 0.3% ciprofloxacin (P<.02). Clinical Relevance Monitoring of laser-induced fluorescence may be helpful in determining the penetration depths and concentrations of topically applied fluoroquinolones within the cornea. PMID- 15534131 TI - Intravitreal voriconazole: an electroretinographic and histopathologic study. AB - BACKGROUND: Voriconazole, a novel triazole antifungal agent, presents potent activity against a broad spectrum of yeast and molds. OBJECTIVE: To determine whether voriconazole could be safely used as an intravitreal agent in the treatment of fungal endophthalmitis. METHODS: Retinal toxicity of voriconazole was examined in a rodent animal model. Voriconazole solutions were serially diluted and injected intravitreally into the eyes of normal adult Sprague-Dawley rats so that the final intravitreal concentrations were 5 microg/mL, 10 microg/mL, 25 microg/mL, 50 microg/mL, and 500 microg/mL (n = 3 for each concentration group). Saline was injected into the fellow eyes of all animals as controls. Three weeks after injections, electroretinograms were measured, and eyes were subsequently enucleated for histologic examination. RESULTS: In electroretinographic studies, maximum scotopic b-wave, intensity needed for half saturation, and saturated a-wave amplitude were measured. There was no statistically significant difference in these parameters recorded between control eyes and voriconazole-injected eyes in any concentration groups. Histologic examination with light microscopy did not reveal any retinal abnormality in the eyes with 5 to 25 microg/mL of intravitreal voriconazole. In the eyes with 50 microg/mL and 500 microg/mL of voriconazole, small foci of retinal necrosis were occasionally observed in the outer retina, especially in the eyes with 500 microg/mL of voriconazole. CONCLUSIONS: Our results demonstrate that intravitreal voriconazole of up to 25 microg/mL [corrected] causes no electroretinographic change or histologic abnormality in rat retinas. This indicates that voriconazole is a safe antifungal agent for intravitreal injection in rodents and may be used in the treatment of human fungal endophthalmitis following further study. PMID- 15534133 TI - Cluster analysis of patients with ocular surface disease, blepharitis, and dry eye. AB - OBJECTIVE: To develop a classification system for blepharitis and dry eye based on a classification-tree model of a large group of subjects who were given a variety of objective physiologic tests. METHODS: We evaluated 513 subjects, some healthy and some with blepharitis and dry eye,with tests for tear volume, tear flow, and tear turnover and the Schirmer test for dry eye. Meibomian gland function was evaluated by meibomian gland lipid expression for lipid volume and lipid viscosity, evaporation, and eyelid transillumination for meibomian gland drop out. We subjected these data to cluster analysis and formulated a classification tree. MAIN OUTCOME MEASURE: The outcome measure of this study was the statistically valid groups of subjects with and without ocular surface symptoms identified by their physiologic characteristics. RESULTS: Cluster analysis most successfully grouped subjects by initially dividing them into 2 groups based on the presence or absence of gland drop out and then by lipid viscosity and volume, Schirmer test results, and evaporation. The analysis created 9 categories. This division created an objective classification system that was found to have clinical relevance. Normal subjects were distributed across several groups. CONCLUSIONS: Using a classification tree, blepharitis and dry eye can be classified with objective physiologic tests into clinically relevant groups that have common characteristics. The analysis establishes the central role of meibomian gland dysfunction in blepharitis and demonstrates the diverse characteristics of the normal population. PMID- 15534134 TI - Now that we have the results of the subretinal surgery trials, how do we manage the patient? PMID- 15534135 TI - Is intensive management of blood pressure to prevent visual loss in persons with type 2 diabetes indicated? PMID- 15534136 TI - Intraretinal calcification and osseous metaplasia in coats disease. PMID- 15534137 TI - Orbital ganglioneuroma in a patient with chronic progressive proptosis. PMID- 15534139 TI - Progression of familial exudative vitreoretinopathy after laser treatment. PMID- 15534138 TI - Primary epithelial-myoepithelial carcinoma of the lacrimal gland. PMID- 15534140 TI - Magnetic resonance imaging of diffuse cerebral vasculitis associated with acute retinal necrosis. PMID- 15534141 TI - Delayed-onset infantile cataracts in a case of Treacher Collins syndrome. PMID- 15534142 TI - Vesicular eruption in a child with trigeminal nerve palsy during topical therapy with substance p and insulinlike growth factor I for neurotrophic keratitis. PMID- 15534143 TI - Retinal hemorrhages as a complication of endoscopic sinus surgery. PMID- 15534144 TI - Bilateral cryptococcal choroiditis. PMID- 15534146 TI - Glaucoma care in a patient with previous anterior ciliary sclerotomy and scleral expansion procedure. PMID- 15534147 TI - Consistency between visual acuity scores obtained at different test distances. PMID- 15534150 TI - An argument against axial length-disc area ratio as the cause of esotropic amblyopia. PMID- 15534152 TI - Diffuse intraocular dispersion of triamcinolone particles as a cause of sterile endophthalmitis. PMID- 15534153 TI - Canaries in the coal mine: The noxious effect of the liability crisis on rural health care: the malpractice crisis becomes a public health crisis. PMID- 15534154 TI - Simple is beautiful: the neglected power of simple tests. PMID- 15534155 TI - Can we ensure that all research subjects give valid consent? AB - To ensure that research subjects provide valid consent, most commentators direct clinical investigators to formally assess potential subjects who are at increased risk for lacking the capacity to consent. Current data reveal, however, that subjects with no known cognitive impairments often fail to give valid consent. These data imply that the prevailing focus on individuals' capacity to consent is too narrow. To protect subjects, as well as the integrity of clinical research, the actual consent of all subjects should be formally assessed. Recent development of several preliminary consent assessment tools suggests that, in addition to being ethically preferable, with additional research this approach may be practically feasible. Future research should focus on developing a postdecision questionnaire that can be adapted to individual studies and used to assess the voluntariness and understanding of all research subjects. PMID- 15534156 TI - Cigarette smoking and infection. AB - BACKGROUND: Infectious diseases may rival cancer, heart disease, and chronic lung disease as sources of morbidity and mortality from smoking. We reviewed mechanisms by which smoking increases the risk of infection and the epidemiology of smoking-related infection, and delineated implications of this increased risk of infection among cigarette smokers. METHODS: The MEDLINE database was searched for articles on the mechanisms and epidemiology of smoking-related infectious diseases. English-language articles and selected cross-references were included. RESULTS: Mechanisms by which smoking increases the risk of infections include structural changes in the respiratory tract and a decrease in immune response. Cigarette smoking is a substantial risk factor for important bacterial and viral infections. For example, smokers incur a 2- to 4-fold increased risk of invasive pneumococcal disease. Influenza risk is severalfold higher and is much more severe in smokers than nonsmokers. Perhaps the greatest public health impact of smoking on infection is the increased risk of tuberculosis, a particular problem in underdeveloped countries where smoking rates are increasing rapidly. CONCLUSIONS: The clinical implications of our findings include emphasizing the importance of smoking cessation as part of the therapeutic plan for people with serious infectious diseases or periodontitis, and individuals who have positive results of tuberculin skin tests. Controlling exposure to secondhand cigarette smoke in children is important to reduce the risks of meningococcal disease and otitis media, and in adults to reduce the risk of influenza and meningococcal disease. Other recommendations include pneumococcal and influenza vaccine in all smokers and acyclovir treatment for varicella in smokers. PMID- 15534157 TI - Impact of the medical professional liability insurance crisis on access to care in Florida. AB - BACKGROUND: Almost half of the US states face serious problems with professional liability insurance (PLI). Despite this, little is known about how this crisis is affecting access to care, particularly in rural areas. METHODS: We surveyed physicians practicing in rural Florida in 2003. The primary assessment was on changes in health care delivery by service type and specialty. Secondary outcomes included changes in PLI premiums and the effect of changes in premiums on service delivery and practice satisfaction. RESULTS: Four hundred eleven (52.6%) of 781 physicians decreased or eliminated health care services during the past year. Overall, 73 (61.3%) of 119 decreased or eliminated vaginal deliveries; 60 (52.6%) of 114, cesarean sections; 186 (51.7%) of 360, hospital-based surgical procedures; 209 (46.4%) of 450, emergency department coverage; 103 (41.7%) of 247, endoscopic procedures; 187 (40.9%) of 457, office-based surgical procedures; and 105 (34.5%) of 304, mental health services. Elimination of services was highest for general surgeons (78.4%), surgical specialists (73.6%), and obstetricians/gynecologists (70.2%). Premiums for PLI rose a mean of 93.5%. Difficulty finding or paying for PLI was listed as an important factor by those reducing or eliminating services and by those planning to leave the community within the next 2 years. CONCLUSIONS: The current crisis in medical PLI in Florida has a major impact on the availability and delivery of health care services to rural areas. Given the number of states that are experiencing similar insurance market upheavals, adverse effects on access to care are likely occurring nationwide. PMID- 15534158 TI - "I wish I had seen this test result earlier!": Dissatisfaction with test result management systems in primary care. AB - BACKGROUND: Failure to review and follow up on outpatient test results in a timely manner represents a patient safety and malpractice concern. Therefore, we sought to identify problems in current test result management systems and possible ways to improve these systems. METHODS: We surveyed 262 physicians working in 15 internal medicine practices affiliated with 2 large urban teaching hospitals (response rate, 64%). We asked physicians about systems they used and the amount of time they spent managing test results. We asked them to report delays in reviewing test results and their overall satisfaction with their management of test results. We also asked physicians to rate features they would find useful in a new test result management system. RESULTS: Overall, 83% of respondents reported at least 1 delay in reviewing test results during the previous 2 months. Despite reporting that they spent on average 74 minutes per clinical day managing test results, only 41% of physicians reported being satisfied with how they managed test results. Satisfaction was associated with fewer self-reported delays in reviewing test results. Physicians who actively tracked their test orders to completion were also more likely to be satisfied. The most highly desired features of a test result management system were tools to help physicians generate result letters to patients, prioritize their workflow, and track test orders to completion. CONCLUSIONS: Delays in test result review are common, and many physicians are not satisfied with how they manage test results. Tools to improve test result management in office practices need to improve workflow efficiency and track test orders to completion. PMID- 15534159 TI - A randomized trial of nortriptyline combined with transdermal nicotine for smoking cessation. AB - BACKGROUND: Smoking cessation rates with current therapy are suboptimal. Tricyclic antidepressants improve cessation rates. We hypothesized that addition of nortriptyline hydrochloride to transdermal nicotine would enhance cessation rates. METHODS: We conducted a randomized, double-blind, placebo-controlled trial at a Department of Veterans Affairs medical center. Subjects were aged 18 to 65 years, smoked 10 or more cigarettes per day, and did not have current major depression. Nortriptyline hydrochloride or matched placebo was started at 25 mg 14 days before quit day, titrated to 75 mg/d as tolerated, and continued for 12 weeks after quit day. Transdermal nicotine (21 mg/d) was started on quit day and continued for 8 weeks. The behavioral intervention consisted of 12 brief, individual visits. Withdrawal symptoms were measured by means of a daily diary, and smoking cessation was defined as self-reported abstinence, expired carbon monoxide level of 9 ppm or less, and a 6-month urine cotinine level less than 50 ng/mL (284 nmol/L). RESULTS: A total of 158 patients were randomized (79 to nortriptyline and 79 to placebo). There was no significant reduction in withdrawal symptoms. The cessation rates at 6 months were 23% (18/79) and 10% (8/79), respectively (absolute difference, 13%; 95% confidence interval, 1.3% 24.5%; P = .052). Nortriptyline caused frequent side effects, including dry mouth (38%) and sedation (20%). CONCLUSIONS: Nortriptyline combined with transdermal nicotine resulted in an increased cessation rate with little effect on withdrawal symptoms. This combination may represent an option for smokers in whom standard therapy has failed. PMID- 15534160 TI - Dietary patterns, meat intake, and the risk of type 2 diabetes in women. AB - BACKGROUND: Although obesity is the most important risk factor for type 2 diabetes, evidence is emerging that certain foods and dietary factors may be associated with diabetes. To examine the association between major dietary patterns and risk of type 2 diabetes mellitus in a cohort of women. METHODS: We prospectively assessed the associations between major dietary patterns and risk of type 2 diabetes in women. Dietary information was collected in 1984, 1986, 1990, and 1994 from 69,554 women aged 38 to 63 years without a history of diabetes, cardiovascular disease, or cancer in 1984. We conducted factor analysis and identified 2 major dietary patterns: "prudent" and "Western." We then calculated pattern scores for each participant and examined prospectively the associations between dietary pattern scores and type 2 diabetes risks. RESULTS: The prudent pattern was characterized by higher intakes of fruits, vegetables, legumes, fish, poultry, and whole grains, while the Western pattern included higher intakes of red and processed meats, sweets and desserts, french fries, and refined grains. During 14 years of follow-up, we identified 2699 incident cases of type 2 diabetes. After adjusting for potential confounders, we observed a relative risk for diabetes of 1.49 (95% confidence interval [CI], 1.26-1.76, P for trend, <.001) when comparing the highest to lowest quintiles of the Western pattern. Positive associations were also observed between type 2 diabetes and red meat and other processed meats. The relative risk for diabetes for every 1 serving increase in intake is 1.26 (95% CI, 1.21-1.42) for red meat, 1.38 (95% CI, 1.23-1.56) for total processed meats, 1.73 (95% CI, 1.39-2.16) for bacon, 1.49 (95% CI, 1.04-2.11) for hot dogs, and 1.43 (95% CI, 1.22-1.69) for processed meats. CONCLUSION: The Western pattern, especially a diet higher in processed meats, may increase the risk of type 2 diabetes in women. PMID- 15534161 TI - Factors influencing the diagnosis of chronic fatigue syndrome. AB - BACKGROUND: Most of what is believed about chronic fatigue syndrome (CFS) is based on clinic-based studies. These studies may not reflect CFS cases in the population. METHODS: We used data from a population-based study of CFS to identify factors associated with receiving a CFS diagnosis. Wichita, Kan, residents were screened by random-digit dialing. Eligible individuals completed a telephone interview. Respondents meeting CFS criteria were invited for a clinical evaluation to confirm CFS. We analyzed all persons with confirmed CFS. The main outcomes of this study, prevalence and incidence of CFS, are published elsewhere. Herein, we present an exploratory analysis with previous CFS diagnosis as the outcome, predicted by demographic and symptom characteristics. RESULTS: We confirmed CFS in 90 subjects; 14 (16%) had been previously diagnosed as having CFS. Persons in the middle- vs the higher-income group were more likely to have been diagnosed as having CFS (9 [29%] of 31 subjects vs 3 [8%] of 39 subjects; P = .03), as were those with sudden vs gradual fatigue onset (7 [41%] of 17 subjects vs 4 [6%] of 64 subjects; P < .01), those reporting tender lymph nodes (7 [33%] of 21 subjects vs 7 [10%] of 69 subjects; P = .02), and those reporting a sore throat (6 [35%] of 17 subjects vs 8 [11%] of 73 subjects; P = .02). Only 17 (21%) of 81 subjects had sudden fatigue onset, and tender lymph nodes (reported in 21 [23%] of 90 subjects) and a sore throat (reported in 17 [19%] of 90 subjects) were the least common symptoms. CONCLUSION: Most cases of CFS in the population are unrecognized by the medical community; persons diagnosed as having CFS may be different from persons with CFS in the general population. PMID- 15534163 TI - Hormone therapy and the impact of estrogen intake on the risk of ovarian cancer. AB - BACKGROUND: The association between menopausal hormone therapy (HT) and risk of ovarian cancer is as yet equivocal, and the effect of estrogen and estrogen progestogen therapy, specifically the effect of the cumulative hormone intake, is unclear. METHODS: We conducted a nationwide population-based case-control study in Denmark. Cases were women aged 35 to 79 years with incident ovarian cancer diagnosed between January 1, 1995, and May 30, 1999. Controls were frequency age matched women from the Danish Central Population Register. The analyses included data on 376 cases who have not undergone hysterectomy and 1111 controls. RESULTS: The risk of ovarian cancer in relation to oral HT increased with the cumulative intake of the estrogen component of HT but not with the duration or the cumulative intake of the progestogen component when the 3 variables were mutually adjusted. A simple trend was found such that each additional gram of estrogen was associated with the same relative increase. The odds ratio was constant throughout the range of cumulative intake. After adjustment for established risk factors, the estimated odds ratio per each additional gram of cumulative estrogen was 1.056 (95% confidence interval, 1.003-1.112), corresponding to an odds ratio of 1.31 (95% confidence interval, 1.01-1.70) per 5 g of estrogen. CONCLUSIONS: Oral HT is associated with risk of ovarian cancer in women who have not undergone hysterectomy. Our results imply that the risk increases with cumulative oral estrogen intake but not with duration of HT, indicating that the increased ovarian cancer risk associated with oral HT may be diminished substantially by minimizing the daily dose of estrogen from oral HT. PMID- 15534162 TI - Relationship between obesity and B-type natriuretic peptide levels. AB - BACKGROUND: The relationships among B-type natriuretic peptide (BNP) levels, body mass index (BMI), and congestive heart failure (CHF) as an emergency diagnosis are unknown. METHODS: Of 1586 participants in the Breathing Not Properly Multinational Study who had acute dyspnea, 1369 (86.3%) had BNP values and self reported height and weight. Two independent cardiologists masked to the BNP results adjudicated the final diagnosis. RESULTS: Congestive heart failure was found in 46% of participants. Individuals with higher BMIs were younger and had more frequent edema on examination but were equally as likely to have CHF vs noncardiac sources of dyspnea. A nearly 3-fold difference was seen in mean +/- SD BNP values at the low and high extremes of the BMI groupings (516.7 +/- 505.9 vs 176.3 +/- 270.5 pg/mL, respectively; P< .001). The correlations between BMI and log BNP among those with and without CHF were r = -0.34 and r = -0.21, respectively (P< .001 for both). Multivariate analysis for the outcome of log BNP among a small subset with CHF (n = 62) found that Framingham score (P = .002), estimated glomerular filtration rate (P = .007), female sex (P = .03), New York Heart Association functional class (P = .09), and third heart sound (P = .08) were independent predictors. However, BMI was not found to be independently related to log BNP (P = .59). CONCLUSIONS: In patients with and without CHF, BNP levels are inversely related to BMI. When considering demographics, severity of disease, and renal function, BMI is not independently related to BNP levels in a small subgroup when detailed information about CHF severity is known. PMID- 15534164 TI - Venous thromboembolism according to age: the impact of an aging population. AB - BACKGROUND: With the aging of the US population, there is concern that the rate of venous thromboembolism will increase, thereby increasing the health burden. In this study we sought to determine trends in the diagnosis of deep venous thrombosis (DVT) and pulmonary embolism (PE) in the elderly as well as the use of diagnostic tests. METHODS: Data from the National Hospital Discharge Survey were used. These data are abstracted each year from a sample of records of patients discharged from non-federal short-stay hospitals in the entire United States. Main outcome measures were trends in rates of diagnosis of DVT and PE as well as trends in the use of diagnostic tests between 1979 and 1999. RESULTS: The rates of diagnosis of DVT and PE and of the use of diagnostic tests over 21 years were markedly higher in elderly than in younger patients (P<.001). Although the rate of diagnosed DVT in elderly patients strikingly increased over the past decade (P< .001), that of PE has been relatively constant. There was a proportionately greater use of venous ultrasonography, ventilation-perfusion lung scanning, and pulmonary angiography in elderly than in younger patients. CONCLUSIONS: Extensive use of diagnostic tests in elderly patients in the past decade has resulted in an increased diagnostic rate for DVT but not PE. The reason for this disparity is uncertain but may reflect early diagnosis and treatment of DVT. With the aging of the population, DVT will increase the health burden. PMID- 15534165 TI - Recurrence risk of oculorespiratory syndrome after influenza vaccination: randomized controlled trial of previously affected persons. AB - BACKGROUND: Oculorespiratory syndrome (ORS) after influenza vaccination has many features of an allergic reaction. METHODS: The objective of the study was to estimate the recurrence rate of ORS after receipt of either of 2 influenza vaccines available in Canada for the 2002-2003 influenza season in individuals who experienced ORS in 2000 or 2001. We designed a randomized, crossover, double blind, placebo-controlled trial in which patients received the vaccine and the placebo 7 days apart. Patients were contacted by telephone at 24 hours and seen at 7 days to collect information about the recurrence of ORS symptoms. The 146 patients belonged to 3 groups: group A (46 patients) had ORS in 2000 but were not revaccinated in 2001, group B (50 patients) had ORS in 2000 and were revaccinated in 2001, and group C (50 patients) had ORS in 2001 but not in 2000. Half of the participants received Fluviral S/F (Shire Biologics) and half received Vaxigrip (Aventis Pasteur). The main outcome measure was the risk difference in ORS symptoms in the 24 hours after receiving the vaccine and after receiving placebo. RESULTS: Recurrence attributable to the vaccine occurred in 34% (95% confidence interval, 21%-47%) of patients after receiving Fluviral S/F and in 15% (95% confidence interval, 2%-28%) after receiving Vaxigrip. The rate was twice as high in group A vs groups B and C. The risk of ORS was highest and most significant in group A patients vaccinated with Fluviral S/F. Most cases were mild, with 94% of patients with recurrence indicating that they would still be revaccinated the next year. CONCLUSIONS: Despite high recurrence rates, revaccination of persons previously affected by ORS seems to be safe. Oculorespiratory syndrome is not anaphylactic, and most recurrences are benign. Most patients remain willing to be revaccinated. PMID- 15534166 TI - Impact of diabetes on mortality in patients with myocardial infarction and left ventricular dysfunction. AB - BACKGROUND: Diabetes is a major risk factor for developing coronary heart disease. In patients with diabetes who survived myocardial infarction (MI), less is known about subsequent morbidity and mortality. We evaluated the effects of diabetes in post-MI patients with left ventricular dysfunction on cardiovascular events and death. METHODS: The Survival and Ventricular Enlargement, a randomized, double-blind, placebo-controlled multicenter trial, evaluated the efficacy of captopril vs placebo in 2231 patients following acute MI with left ventricular dysfunction defined as an ejection fraction less than or equal to 40%. Patients were randomly assigned to captopril or placebo 3 to 16 days following MI and were followed up for 2 to 5 years (mean, 3.5 years). RESULTS: Among the 2231, 496 (22.2%) were patients with a history of diabetes, of which 168 (33.9%) were treated with insulin. Patients with diabetes were significantly older; more likely to be women; have a history of prior MI or hypertension; be obese or manifest Killip class II or greater; and have higher systolic blood pressure, pulse pressure, and heart rate, as well as lower ejection fraction. During follow-up, 31.3% of patients with diabetes and 20.1% of nondiabetic patients died (P<.001). Furthermore, 50% of the patients with diabetes had at least 1 major cardiovascular event compared with 32.3% among the nondiabetic patients (P<.001). In multivariate analysis that adjusted for all significant differences in baseline characteristics, patients with diabetes had a 39% higher total mortality (P = .001) and 49% more cardiovascular events (P = .001). Among the patients with diabetes, baseline insulin treatment was associated with a greater risk of death (41.1% vs 26.2%; P = .001) and cardiovascular events (58.3% vs 45.7%; P = .008). CONCLUSIONS: In patients who survived MI with left ventricular dysfunction, diabetes increased risk of death from all causes even after controlling for differences in other risk factors. Patients with diabetes treated with insulin have a particularly higher mortality risk. Patients with diabetes who survived MI with left ventricular dysfunction, in particular those receiving insulin, are at high risk of subsequent mortality and cardiovascular events and thus require intensive risk factor modification, as well as evaluation for novel therapies. PMID- 15534168 TI - The danger of single-payer health insurance. PMID- 15534169 TI - A single-payer national health insurance: we gave twice at the office. PMID- 15534170 TI - Are cola drinkers at risk of hypovitaminosis C? PMID- 15534171 TI - Establishing an optimal therapeutic range for coumarins. PMID- 15534172 TI - Antisperm antibodies in inflammatory bowel disease. PMID- 15534173 TI - SARS, SIRS, and neurological disease. PMID- 15534174 TI - Tales from the neural genome: the lessons of homozygous porphyria. PMID- 15534175 TI - Meta-analysis of genetic studies in ischemic stroke: thirty-two genes involving approximately 18,000 cases and 58,000 controls. AB - Ischemic stroke is thought to have a polygenic basis, but identification of stroke susceptibility genes and quantification of associated risks have been hampered by conflicting results from underpowered case-control studies. We performed a meta-analysis of all candidate gene association studies in ischemic stroke. Electronic databases were searched up until January 2003 for all case control and nested case-control studies in English-language journals relating to the investigation of any candidate gene for ischemic stroke in humans. Cases were required to have neuroimaging evidence of the diagnosis. To maintain genetic homogeneity, only studies in white adults were included. Studies that evaluated quantitative traits or intermediate phenotypes were excluded. Data from 120 case control studies were included. Pooled odds ratios (ORs) with 95% confidence intervals (CIs) from random- and fixed-effects models were calculated. Of 32 genes studied, 15 polymorphisms were identified for which at least 3 studies had been conducted. Statistically significant associations with ischemic stroke were identified for factor V Leiden Arg506Gln (OR, 1.33; 95% CI, 1.12-1.58), methylenetetrahydrofolate reductase C677T (OR, 1.24; 95% CI, 1.08-1.42), prothrombin G20210A (OR, 1.44; 95% CI, 1.11-1.86), and angiotensin-converting enzyme insertion/deletion (OR, 1.21; 95% CI, 1.08-1.35). These were also the most investigated candidate genes, including 4588, 3387, 3028, and 2990 cases, respectively. No statistically significant association with ischemic stroke was detected for the 3 next most investigated genes (factor XIII, apolipoprotein E, and human platelet antigen type 1). There is a genetic component to common stroke. No single gene with major effect was identified; rather, common variants in several genes, each exerting a modest effect, contribute to the risk of stroke. These findings have important implications for the design of future genetic studies and for predictive genetic testing for stroke and other multifactorial diseases. PMID- 15534176 TI - Channels and disease: past, present, and future. PMID- 15534177 TI - Neuromuscular disorders in severe acute respiratory syndrome. AB - OBJECTIVE: To delineate and clarify neuromuscular disorders in patients with probable severe acute respiratory syndrome (SARS). DESIGN: Case series with follow-up ranging from 3 weeks to 2 months. SETTING: National Taiwan University Hospital, Taipei. PATIENTS: We investigated 4 patients with SARS who had concomitant neuromuscular problems. A diagnosis of SARS was based on the demonstration of serum coronavirus antibodies. Clinical presentations, laboratory results, electrophysiologic findings, and follow-up conditions were determined. RESULTS: Patients developed neuromuscular problems approximately 3 weeks after the onset of SARS. Two women experienced motor-predominant peripheral nerve disorders. A man developed myopathy and a third woman experienced neuropathy and myopathy. Cerebrospinal fluid obtained from 2 patients with neuropathy disclosed normal protein content and the absence of pleocytosis and SARS coronavirus antibodies. Both patients with myopathy had elevated serum creatine kinase levels. A rapid clinical and electrophysiologic improvement was evident during follow-up examinations, with a good prognosis. CONCLUSIONS: The neuromuscular problems in patients with SARS are considered to be critical-illness polyneuropathy or myopathy, possibly coexistent. Further pathological and microbiological studies are necessary to determine the relationship between SARS coronavirus and neuromuscular problems. PMID- 15534178 TI - Factor analysis of the National Institutes of Health Stroke Scale in patients with large strokes. AB - BACKGROUND: The National Institutes of Health Stroke Scale (NIHSS) was created to detect treatment-related differences in clinical trials and was designed to measure right- and left-sided cerebral hemispheric function. OBJECTIVE: To validate the original design in patients with very large strokes. DESIGN: A previously published factor structure was fit to the data. Then, a new analysis was conducted to explore the underlying structure of the scale in this population. Finally, NIHSS scores and infarction volumes were compared. SETTING: The Clomethiazole for Acute Stroke Study-Ischemic, conducted in academic and community hospitals. PATIENTS: Individuals with acute stroke seen within 12 hours of onset. Of 1191 records available, 98% had complete NIHSS scores. MAIN OUTCOME MEASURE: Goodness-of-fit statistic (Bentler) for each factor solution. RESULTS: Two factors were found underlying the NIHSS, corresponding to the left and right hemispheres (goodness of fit = 0.97), using the previously published factor analysis. The new exploratory analysis also suggested 2 factors representing left and right brain function. The median (range) NIHSS scores were 15 (5-25) for right brain strokes and 19 (6-32) for left brain strokes (P<.001). The median (range) infarction volumes were 56.2 mL (0.1-381.5 mL) for right brain strokes and 37.8 mL (0.2-255.1 mL) for left brain strokes (P<.001). The correlation coefficient between NIHSS score and lesion volume was 0.37 (P<.001). CONCLUSIONS: The underlying structure of the NIHSS conforms to cerebral hemispheric lateralization, confirming previous findings in a new population of large hemispheric strokes. Left- brain strokes score 4 points higher on the NIHSS than right brain strokes of larger volume. PMID- 15534179 TI - Factors affecting the angiographic recanalization and early clinical improvement in middle cerebral artery territory infarction after thrombolysis. AB - BACKGROUND: Factors affecting the angiographic recanalization (AR) and clinical improvement (CI) still remain unclear in patients receiving thrombolytic therapy. OBJECTIVES: To elucidate factors related to AR and early CI in patients with middle cerebral artery (MCA) or internal carotid artery (ICA) occlusion. DESIGNS: Retrospective study. SETTING: Department of Neurology, Asan Medical Center, Seoul, South Korea. PATIENTS: We studied 42 patients who (1) underwent diffusion weighted magnetic resonance (MR) imaging and MR angiography within 6 hours after onset, (2) had MCA territory infarction, (3) had nonvisualization of the MCA or the ICA on initial MR angiography, (4) were treated with thrombolytics, and (5) underwent follow-up MR imaging and MR angiography at day 2 or 3. RESULTS: Successful AR and CI were achieved in 31 and 16 patients, respectively. Angiographic recanalization was related to CI (P<.01), lower follow-up National Institutes of Health Stroke Scale scores (P<.05), the absence of a dominant ipsilateral posterior cerebral artery (P<.01) on initial MR angiography, and the sparing of the internal capsule on both initial (P<.05) and follow-up (P<.01) MR imaging. Clinical improvement was associated with the absence of ICA (vs MCA) flow signals (P<.05), the sparing of the internal capsule (P<.01), and marginally, with the infarct volume change (P = .06). CONCLUSIONS: In patients with MCA or ICA occlusion, CI after thrombolysis is related to the AR and the sparing of the critical motor pathway. The presence of a dominant ipsilateral posterior cerebral artery may predict poor AR after thrombolysis. PMID- 15534181 TI - Antiretroviral therapy in HIV infection: are neurologically active drugs important? AB - BACKGROUND: The effect on neuropsychological function of antiretroviral drugs that are able to penetrate into the brain in effective concentration (neuroactive drugs) remains unclear. OBJECTIVE: To investigate whether highly active antiretroviral therapy (HAART) containing neuroactive drugs is associated with better neuropsychological performance in patients with human immunodeficiency virus disease. DESIGN: Cross-sectional survey. SETTING: Tertiary referral hospital outpatient clinics. PATIENTS: The study population consisted of 97 individuals positive for human immunodeficiency virus (stage C3, 1993 Centers for Disease Control and Prevention classification) whose condition had been stable on their current HAART regimen for a mean +/- SD of 18.5 +/- 16.5 months and who were aged 48.14 +/- 9.38 years. The patient groups were analyzed according to whether their regimen contained 3 or more neuroactive drugs (neuroHAART group; n = 41) or not (HAART group; n = 56). Thirty seronegative men matched for age and education were recruited as controls. MAIN OUTCOME MEASURE: Neuropsychological performance on 7 cognitive domains. RESULTS: The neuroHAART and HAART groups did not differ from one another on neuropsychological performance, but both patient groups were impaired compared with controls. Impaired patients in each treatment group were compared, and the neuroHAART group showed significantly better memory performance, unrelated to plasma viral load, than the HAART group. CONCLUSION: No direct benefit of neuroactive HAART therapy was found in patients with advanced human immunodeficiency virus infection. However, in neuropsychologically impaired patients, there was a benefit in memory function. This suggests that a threshold of neuropsychological impairment is required for the benefit of neuroactive HAART. PMID- 15534180 TI - Attenuated central nervous system infection in advanced HIV/AIDS with combination antiretroviral therapy. AB - BACKGROUND: Before the introduction of combination antiretroviral therapy (CART), neurological disease correlated with cerebrospinal fluid (CSF) levels of human immunodeficiency virus (HIV) RNA. OBJECTIVE: To investigate the relationships among HIV RNA levels, immune activation markers, and neurological status in patients receiving CART. DESIGN: Multicenter cohort study. SETTING: Academic neurology departments. PATIENTS: A total of 371 patients unselected for neurological complaints and with CD4 cell counts less than 200/microL or with cognitive symptoms and CD4 cell counts less than 300/microL were enrolled into the Northeastern AIDS Dementia cohort in 1998-2002. Diagnoses of HIV-associated dementia (HIV-D) and minor cognitive-motor disorder (MCMD) were obtained with a computerized algorithm. Plasma and CSF levels of HIV RNA, monocyte chemotactic protein 1, macrophage colony-stimulating factor, and tumor necrosis factor alpha were quantified. RESULTS: The mean +/- SD age was 41.5 +/- 7.2 years, and the mean +/- SD educational level was 12.3 +/- 2.2 years. Seventy percent of the cohort was black, and 30% were women. The mean +/- SD CD4 cell count was 136.8 +/ 87.9/microL, and CART was used in 71%. Twenty-nine percent of the patients were unimpaired (n = 106), 36% had MCMD (n = 133), and 35% had HIV-D (n = 128). Mean log(10) CSF HIV RNA copies per milliliter was 2.6 +/- 0.8, with no differences among the neurological groups, even after adjustments for baseline CD4 cell counts and antiretroviral therapy. Cerebrospinal fluid HIV RNA was undetectable in 47% of unimpaired, 46% of MCMD, and 43% of HIV-D patients (P = .91). Plasma levels of monocyte chemotactic protein type 1 and tumor necrosis factor alpha correlated weakly with HIV RNA levels but did not distinguish those with neurological deficits. CONCLUSIONS: In contrast to observations in individuals not treated with CART, we found no relationship between CSF markers and neurological status in this CART-using cohort with advanced HIV/AIDS. This was not explicable by demographic differences or plasma virological control. CART may substantially attenuate the degree of central nervous system HIV infection and immune activation, and in CART users, CSF HIV RNA and immune activation markers may fail to discriminate milder degrees of HIV-D and MCMD. PMID- 15534182 TI - Role of dopaminergic treatment in dopamine receptor down-regulation in advanced Parkinson disease: a positron emission tomographic study. AB - BACKGROUND: In patients with advanced Parkinson disease (PD) who are undergoing long-term treatment with a dopaminergic medication, a down-regulation of striatal dopamine D2 receptor expression has been demonstrated and interpreted as a consequence of either the disease itself or dopaminergic drug administration. OBJECTIVE: To compare, using positron emission tomography, the striatal binding of raclopride carbon C 11, a dopamine D2 receptor ligand, in PD patients who completely discontinued dopaminergic therapy (off drug) with that in PD patients who continued receiving dopaminergic therapy (on drug) after undergoing subthalamic nucleus stimulation. MAIN OUTCOME MEASURES: The positron emission tomographic data were acquired in off-stimulation and, for 12 hours, off medication conditions. Five off-drug PD patients, 7 on-drug PD patients, and 8 healthy subjects participated. RESULTS: In off-drug PD patients, the putaminal raclopride C 11 binding was 24% higher than in on-drug PD patients. The same tendency was noted for the caudate nucleus, but was not significant (P=.07). Compared with control subjects, the putaminal raclopride C 11 binding was increased by 21% in off-drug and was normal in on-drug PD patients. Compared with controls, the caudate raclopride C 11 binding was reduced by 23% in on-drug and was normal in off-drug PD patients. Further analysis using statistical parametric mapping showed a significant increase of binding bilaterally in the caudate nucleus and putamen in off-drug compared with on-drug PD patients (P=.002 at cluster level). CONCLUSIONS: The down-regulation of dopamine D2 receptors probably relates to the long-term and intermittent administration of dopaminergic treatments rather than to disease progression. This phenomenon is reversed by the complete withdrawal of dopaminergic drugs. Furthermore, an up-regulation of putaminal dopamine D2 receptors is demonstrated in late-stage PD after dopaminergic drug withdrawal. PMID- 15534183 TI - Reduced brain delivery of homovanillic acid to cerebrospinal fluid during human aging. AB - BACKGROUND: Markers of human brain dopamine metabolism are reported to decline with age. However, the cerebrospinal fluid (CSF) concentration of homovanillic acid (HVA), a major dopamine metabolite, is reported to not change or to increase in elderly individuals. OBJECTIVE: To estimate the rate of delivery of HVA from the brain to CSF, taking into account the HVA concentration gradient in the spinal subarachnoid space and CSF flow. METHODS: Homovanillic acid concentrations were measured in 5 serial 6-mL aliquots of CSF removed from the L3-4 or L4-5 interspaces of 7 healthy young (mean +/- SD age, 28.7 +/- 4.6 years) subjects and 7 healthy elderly (mean +/- SD age, 77.1 +/- 6.3 years) subjects. Cisterna magna HVA concentrations were estimated from the slopes of the HVA concentrations along the spinal subarachnoid space. The products of cisternal HVA concentrations and published values for CSF flow were used to estimate lower limits for brain delivery of HVA to CSF, according to the Fick principle. RESULTS: The mean +/- SD HVA concentration in the initial lumbar CSF sample in the young subjects, 116 +/- 66 pmol/mL, did not differ significantly from 140 +/- 86 pmol/mL in the elderly subjects. Estimated cisternal HVA concentrations equaled 704 and 640 pmol/mL, respectively, in the young and elderly subjects. Multiplying these concentrations by CSF flow rates of 591 and 294 mL/d, respectively, gave lower limits for rates of delivery of HVA from the brain to CSF. These rates equaled 416 and 175 nmol/d, respectively. CONCLUSION: A 50% decline in the lower limit for the rate of HVA delivery from the brain to CSF in elderly individuals is consistent with other evidence that brain dopaminergic neurotransmission declines with age. PMID- 15534184 TI - Medial temporal hypoperfusion and aggression in Alzheimer disease. AB - BACKGROUND: It is not understood why some patients with Alzheimer disease (AD) display aggression and others do not. OBJECTIVE: To examine the relation between regional brain perfusion and aggression in AD. DESIGN: Single-photon emission computed tomographic scans were coregistered to a standardized template in Talairach space, generating mean ratios of uptake referenced to the cerebellum. PARTICIPANTS: Forty-nine outpatients (25 men and 24 women; mean +/- SD age, 74 +/ 11 years) with probable AD (Mini-Mental State Examination score, 17.7 +/- 5.0; 30 aggressive and 19 nonaggressive), comparable in age, sex, and severity of cognitive impairment. MAIN OUTCOME MEASURES: Regional perfusion ratios were determined for 5 bilateral regions of interest: orbitofrontal, middle medial temporal, inferior medial temporal, hypothalamus/thalamus, and anterior cingulate. RESULTS: Compared with nonaggressive patients, aggressive ones displayed hypoperfusion in the right and left middle medial temporal regions of interest (P = .02 for both), but not the others (all (t tests, unpaired, 2 tailed). On regression analyses, right middle temporal hypoperfusion (P = .001), younger age (P = .002), greater activity disturbances (P = .004), and higher Mini Mental State Examination scores (P = .04) independently predicted aggression, accounting for 44% of the total variance (F = 8.7; P<.001). Statistical parametric mapping analyses supported right middle medial temporal hypoperfusion in the aggressive group (P = .008). CONCLUSION: In this sample of patients with AD, the right middle medial temporal region emerged as an important neural correlate of aggression. PMID- 15534185 TI - The natural history of Alzheimer disease: a longitudinal presymptomatic and symptomatic study of a familial cohort. AB - BACKGROUND: Knowledge of the evolution of cognitive deficits in Alzheimer disease is important for our understanding of disease progression. Previous reports, however, have either lacked detail or have not covered the presymptomatic stages. OBJECTIVE: To delineate the onset and progression of clinical and neuropsychological abnormalities in familial Alzheimer disease. METHODS: Nineteen subjects with familial Alzheimer disease underwent serial clinical and neuropsychological assessments. Eight of these had undergone presymptomatic assessments. The follow-up period was 1 to 10 years (mean, 5 years). The relative timing of the occurrence of 3 markers of disease onset and progression (onset of symptoms, Mini-Mental State Examination score < or = 24, and impaired scores on a range of neuropsychological tests) were compared using the binomial exact test. RESULTS: Neurological abnormalities were not prominent, although myoclonus appeared early in some. Mini-Mental State Examination score was not sensitive to early disease. Memory and general intelligence deficits appeared at an earlier stage, in some patients when presymptomatic. Perceptual, naming, and especially spelling skills were preserved to a late stage. CONCLUSION: Familial Alzheimer disease may have a long prodromal phase of several years with subtle deficits initially of general intelligence and memory, while spelling, naming, and perception are relatively preserved until a late stage. PMID- 15534186 TI - Cognitive impairments in Machado-Joseph disease. AB - BACKGROUND: Cognitive function of Machado-Joseph disease (MJD) patients has not been clarified. OBJECTIVES: To determine the characteristics of cognitive dysfunction in MJD patients and to assess the relationship of dysfunction to age at onset, age at examination, disease duration, education, ataxia, depression, anxiety, and CAG repeat length. DESIGN: Case-control study. SETTING: Research oriented hospitals. PARTICIPANTS: Sixteen genetically confirmed MJD patients able to complete neuropsychological tests and 20 control subjects matched to patients by age and education. MAIN OUTCOME MEASURES: Neuropsychological tests, including general cognition, verbal and visual memory, working memory, visuospatial and constructional ability, language, executive function, depression, and anxiety. RESULTS: Machado-Joseph disease patients scored significantly lower than controls in verbal and visual memory, in visuospatial and constructional tasks, and in phonemic and semantic fluency tasks. None of these impairments correlated with CAG repeat length, age at onset, age at examination, disease duration, or education. Verbal fluency (words named in a category) correlated with the International Cooperative Ataxia Rating Scale score. CONCLUSION: Machado-Joseph disease patients have verbal and visual memory deficits, visuospatial and constructional dysfunction, and verbal fluency deficits, all unrelated to CAG repeat length. PMID- 15534187 TI - Acute intermittent porphyria: studies of the severe homozygous dominant disease provides insights into the neurologic attacks in acute porphyrias. AB - BACKGROUND: Acute intermittent porphyria (AIP), due to half-normal hydroxymethylbilane synthase activity,is characterized by acute life-threatening neurologic attacks whose etiology remains unclear. To date, only 3 patients confirmed to have homozygous dominant AIP (HD-AIP) have been described (hydroxymethylbilane synthase genotypes R167Q/R167Q and R167W/R173Q). OBJECTIVE: To investigate the genetic, biochemical, clinical, and neuroradiologic features of a severely affected infant with HD-AIP. DESIGN: Clinical, imaging, and genotype/phenotype studies were performed. RESULTS: The proband, homoallelic for hydroxymethylbilane synthase mutation R167W, had approximately 1% of normal hydroxymethylbilane synthase activity, elevated porphyrins and porphyrin precursors, severe psychomotor delay, and central and peripheral neurologic manifestations. When expressed in vitro, the R167W mutant enzyme had less than 2% of normal activity but was markedly unstable, consistent with the proband's severe phenotype. Mitochondrial respiratory chain enzymes were normal. Neuroradiologic studies revealed a unique pattern of deep cerebral white matter injury, with relative preservation of the corpus callosum, anterior limb of the internal capsule, cerebral gray matter, and infratentorial structures. CONCLUSIONS: This severely affected patient with HD-AIP expanded the phenotypic spectrum of HD-AIP. His brain magnetic resonance imaging studies suggested selective cerebral oligodendrocyte postnatal involvement in HD-AIP, whereas most structures developed prenatally were intact. These findings indicate that the neurologic manifestations result from porphyrin precursor toxicity rather than heme deficiency and suggest that porphyrin precursor toxicity is primarily responsible for the acute neurologic attacks in heterozygous AIP and other porphyrias. PMID- 15534188 TI - A novel presenilin-1 mutation (Leu85Pro) in early-onset Alzheimer disease with spastic paraparesis. AB - BACKGROUND: Early-onset familial Alzheimer disease is caused by mutations in the amyloid precursor protein (APP), presenilin-1 (PSEN1), or presenilin-2 (PSEN2) genes. Phenotypic diversity has been reported to be associated with various mutations in PSEN1. Various mutations of PSEN1 have been reported in cases of early-onset Alzheimer disease with spastic paraparesis. OBJECTIVE: To describe a novel mutation in the PSEN1 gene associated with early-onset Alzheimer disease with spastic paraparesis. PATIENT AND METHODS: The patient was a 27-year-old man who developed early-onset dementia with spastic paraparesis. We examined sequences of the PSEN1, PSEN2, and APP genes from the patient and his family. To detect a possible mutation effect on the production of amyloid-beta peptide (Abeta), transfected HEK293 cells were examined for Abeta42 and Abeta40 production. RESULTS: We found a novel mutation (Leu85Pro) in PSEN1. This mutation influenced the production of Abeta, resulting in a 2-fold elevation of Abeta42 production and of the Abeta42/40 ratio. CONCLUSION: To our knowledge, this is the first report of very early-onset Alzheimer disease with spastic paraparesis and with the visual variant form of the disease, which is associated with visuospatial cognitive disorder. The Leu85Pro mutation in PSEN1 was pathogenic. PMID- 15534189 TI - A novel polymerase gamma mutation in a family with ophthalmoplegia, neuropathy, and Parkinsonism. AB - BACKGROUND: Mutations in polymerase gamma cause progressive external ophthalmoplegia and a variety of associated symptoms and signs, including neuropathy, ataxia, hypogonadism, hearing loss, muscle weakness, and psychiatric problems. Extrapyramidal signs have been rarely described. OBJECTIVE: To describe a family with a novel polymerase gamma mutation and autosomal dominant transmission of progressive external ophthalmoplegia, neuropathy, hypogonadism, and parkinsonism. DESIGN: Case report. PATIENTS: The proband, a 49-year-old woman with incipient parkinsonism, and her 59-year-old brother with overt parkinsonian features. MAIN OUTCOME MEASURES: Mutation in the proband by sequencing the polymerase gamma gene and in affected relatives by restriction fragment length polymorphism analysis. RESULTS: We found multiple mitochondrial DNA deletions in the proband's muscle and a novel missense mutation in the polymerase gamma gene (A2492G) in the proband and in her affected siblings. CONCLUSION: Parkinsonism was a prominent clinical feature in this family with autosomal dominant ophthalmoplegia, multiple mitochondrial DNA deletions, and a novel mutation in the polymerase gamma gene. PMID- 15534191 TI - Ring enhancement after hemorrhagic stroke. PMID- 15534190 TI - Serial 18F-fluoro-2-deoxy-D-glucose positron emission tomography and magnetic resonance imaging of paraneoplastic limbic encephalitis. AB - OBJECTIVES: To review and expand the existing literature of magnetic resonance imaging (MRI) and positron emission tomography (PET) of paraneoplastic limbic encephalitis (PLE). METHODS: We performed serial MRI and 18F-fluoro-2-deoxy-D glucose (FDG)-PET in a patient with anti-Ma2-positive PLE. In addition, we reviewed the relevant literature by conducting a search in the Medline database. RESULTS: We found a total of 7 published patient studies of possible or probable PLE containing both MRI and PET data. In 1 of these reports, the diagnosis of PLE can be regarded as proven. The results of the previous studies are controversial. Epileptic activity and inflammation are assumed to be underlying mechanisms of increased FDG uptake. In our study, we found a focal tracer accumulation in the left medial temporal lobe, which increased during the first 9 months of follow-up and corresponded with an increase of serum anti-Ma2 antibody titers. The MRI findings showed a hyperintense signal change in the left medial temporal lobe without contrast enhancement, which remained unchanged over time. CONCLUSIONS: The results of functional and structural imaging in PLE may differ substantially. Results of FDG-PET can demonstrate focal hypermetabolism over a long time, which may indicate therapeutic potential. A prospective study with more patients will be needed to clarify the relevance of PET as a possible outcome measure in PLE. Future studies should include scalp or semi-invasive electroencephalographic recordings during PET acquisition. PMID- 15534192 TI - Multiple central nervous system lesions associated with rheumatoid arthritis. PMID- 15534193 TI - Psychiatric symptoms and brain tumors: a brief historical overview. PMID- 15534194 TI - Quality of life is not favorable for most patients with multiple sclerosis. PMID- 15534195 TI - Adaptation is favorable for most patients with multiple sclerosis. PMID- 15534196 TI - Positive potential of fetal nigral implants for Parkinson disease. PMID- 15534197 TI - Biography of Peter H. Quail. PMID- 15534198 TI - Adeno-associated viral vector delivers cardiac-specific and hypoxia-inducible VEGF expression in ischemic mouse hearts. AB - It has been shown that the adeno-associated virus (AAV) vector can deliver the VEGF gene efficiently into the ischemic mouse myocardium. However, the AAV genomes can be found in extracardiac organs after intramyocardial injection. To limit unwanted VEGF expression in organs other than the heart, we tested the use of the cardiac myosin light chain 2v (MLC-2v) promoter and the hypoxia-response element to mediate cardiac-specific and hypoxia-inducible VEGF expression. An AAV vector, MLCVEGF, with 250 bp of the MLC-2v promoter and nine copies of the hypoxia-response element driving VEGF expression, was constructed. Gene expression was studied in vitro by infection of rat cardiomyocytes, rat skeletal myocytes, and mouse fibroblasts with the vector and in vivo by direct injection of the vector into normal and ischemic mouse hearts. With MLCVEGF infection, VEGF expression was higher in cardiomyocytes than the other two cell lines and was hypoxiainducible. VEGF expression was also higher in ischemic hearts than in normal hearts. No VEGF expression was detectable in organs with detectable MLCVEGF vectors other than the heart. MLCVEGF-injected ischemic hearts had more capillaries and small vessels around the injection site, smaller infarct size, and better cardiac function than the negative controls. Hence, MLCVEGF can mediate cardiac-specific and hypoxia-inducible VEGF expression, neoangiogenesis, infarct-size reduction, and cardiac functional improvement. PMID- 15534199 TI - Long-term meditators self-induce high-amplitude gamma synchrony during mental practice. AB - Practitioners understand "meditation," or mental training, to be a process of familiarization with one's own mental life leading to long-lasting changes in cognition and emotion. Little is known about this process and its impact on the brain. Here we find that long-term Buddhist practitioners self-induce sustained electroencephalographic high-amplitude gamma-band oscillations and phase synchrony during meditation. These electroencephalogram patterns differ from those of controls, in particular over lateral frontoparietal electrodes. In addition, the ratio of gamma-band activity (25-42 Hz) to slow oscillatory activity (4-13 Hz) is initially higher in the resting baseline before meditation for the practitioners than the controls over medial frontoparietal electrodes. This difference increases sharply during meditation over most of the scalp electrodes and remains higher than the initial baseline in the postmeditation baseline. These data suggest that mental training involves temporal integrative mechanisms and may induce short-term and long-term neural changes. PMID- 15534200 TI - Reversible oxidation and inactivation of the tumor suppressor PTEN in cells stimulated with peptide growth factors. AB - Stimulation of cells with various peptide growth factors induces the production of phosphatidylinositol 3,4,5-trisphosphate (PIP3) through activation of phosphatidylinositol 3-kinase. The action of this enzyme is reversed by that of the tumor suppressor PTEN. With the use of cells overexpressing NADPH oxidase 1 or peroxiredoxin II, we have now shown that H2O2 produced in response to stimulation of cells with epidermal growth factor or platelet-derived growth factor potentiates PIP3 generation and activation of the protein kinase Akt induced by these growth factors. We also show that a small fraction of PTEN molecules is transiently inactivated as a result of oxidation of the essential cysteine residue of this phosphatase in various cell types stimulated with epidermal growth factor, platelet-derived growth factor, or insulin. These results suggest that the activation of phosphatidylinositol 3-kinase by growth factors might not be sufficient to induce the accumulation of PIP3 because of the opposing activity of PTEN and that the concomitant local inactivation of PTEN by H2O2 might be needed to increase the concentration of PIP3 sufficiently to trigger downstream signaling events. Furthermore, together with previous observations, our data indicate that peroxiredoxin likely participates in PIP3 signaling by modulating the local concentration of H2O2. PMID- 15534201 TI - Fly motion vision is based on Reichardt detectors regardless of the signal-to noise ratio. AB - The computational structure of an optimal motion detector was proposed to depend on the signal-to-noise ratio (SNR) of the stimulus: At low SNR, the optimal motion detector should be a correlation or "Reichardt" type, whereas at high SNR, the detector would employ a gradient scheme [Potters, M. & Bialek, W. (1994) J. Physiol. (Paris) 4, 1755-1775]. Although a large body of experiments supports the Reichardt detector as the processing scheme leading to direction selectivity in fly motion vision, in most of these studies the SNR was rather low. We therefore reinvestigated the question over a much larger SNR range. Using 2-photon microscopy, we found that local dendritic [Ca(2+)] modulations, which are characteristic of Reichardt detectors, occur in response to drifting gratings over a wide range of luminance levels and contrasts. We also explored, as another fingerprint of Reichardt detectors, the dependence of the velocity optimum on the pattern wavelength. Again, we found Reichardt-typical behavior throughout the whole luminance and contrast range tested. Our results, therefore, provide strong evidence that only a single elementary processing scheme is used in fly motion vision. PMID- 15534202 TI - Crystal structure of a human CD3-epsilon/delta dimer in complex with a UCHT1 single-chain antibody fragment. AB - The alpha/beta T cell receptor complex transmits signals from MHC/peptide antigens through a set of constitutively associated signaling molecules, including CD3-epsilon/gamma and CD3-epsilon/delta. We report the crystal structure at 1.9-A resolution of a complex between a human CD3-epsilon/delta ectodomain heterodimer and a single-chain fragment of the UCHT1 antibody. CD3 epsilon/delta and CD3-epsilon/gamma share a conserved interface between the Ig fold ectodomains, with parallel packing of the two G strands. CD3-delta has a more electronegative surface and a more compact Ig fold than CD3-gamma; thus, the two CD3 heterodimers have distinctly different molecular surfaces. The UCHT1 antibody binds near an acidic region of CD3-epsilon opposite the dimer interface, occluding this region from direct interaction with the TCR. This immunodominant epitope may be a uniquely accessible surface in the TCR/CD3 complex, because there is overlap between the binding site of the UCHT1 and OKT3 antibodies. Determination of the CD3-epsilon/delta structure completes the set of TCR/CD3 globular ectodomains and contributes information about exposed CD3 surfaces. PMID- 15534203 TI - Reversion of the Jun-induced oncogenic phenotype by enhanced synthesis of sialosyllactosylceramide (GM3 ganglioside). AB - In the mouse fibroblast cell line C3H 10T1/2 and the chicken fibroblast cell line DF1, the ganglioside GM3 is the major glycosphingolipid component of the plasma membrane. Expression of the viral oncoprotein Jun (v-Jun) induces transformed cell clones with greatly reduced levels of GM3 and GM3 synthase (lactosylceramide alpha2,3-sialyltransferase) mRNA in both 10T1/2 and DF1 cell cultures. Compared with nontransformed controls, v-Jun transfectants show enhanced ability of anchorage-independent growth, and their growth rates as adherent cells are increased. When the mouse GM3 synthase gene is transfected with the pcDNA vector into v-Jun-transformed 10T1/2 cells, the levels of GM3 synthase and corresponding mRNA are restored to those of control cells. Reexpression of GM3 correlates with a reduced ability of the cells to form colonies in nutrient agar. Similarly, when the newly cloned chicken GM3 synthase gene is transfected into v-Jun-transformed DF1 with the pcDNA vector, the GM3 synthase level is restored to that of control cells, and the ability of the cells to form agar colonies is reduced. The levels of GM3 in the cell also affect membrane microdomains. The complex of GM3 with tetraspanin CD9 and integrin alpha5beta1 inhibits motility and invasiveness. The amounts of this complex are greatly reduced in transformed cells. Expression of GM3 and consequent reversion of the transformed phenotype results in increased levels of that microdomain complex. PMID- 15534204 TI - Alerting dendritic cells to pathogens: the importance of Toll-like receptor signaling of stromal cells. PMID- 15534205 TI - Genes required for Drosophila nervous system development identified by RNA interference. AB - RNA interference was used to screen 3,314 Drosophila double-stranded RNAs, corresponding to approximately 25% of Drosophila genes, for genes that affect the development of the embryonic nervous system. RNA-interference-mediated gene silencing in Drosophila embryos resulted in loss-of-function mutant phenotypes for 43 genes, which is 1.3% of the genes that were screened. We found 18 genes that were not known previously to affect the development of the nervous system. The functions of some of the genes are unknown. Other genes encode protein kinases, transcription factors, and signaling proteins, as well as proteins with other functions. PMID- 15534206 TI - Evolving protein functional diversity in new genes of Drosophila. AB - The mechanism by which protein functional diversity expands is an important evolutionary issue. Studies of recently evolved chimeric genes permit direct investigation of the origin of new protein functions before they become obscured by subsequent evolution. Found in several African Drosophila species, jingwei (jgw), a recently evolved gene with a domain derived from the still extant short chain alcohol dehydrogenase (ADH) through retroposition, provides an opportunity to examine this previously undescribed process directly. We expressed JGW proteins in a microbial expression system and, after purification, investigated their enzymatic properties. We found that, unexpectedly, positive Darwinian selection for amino acid replacements outside the active site of JGW produced a novel dehydrogenase with altered substrate specificity compared with the ancestral ADH. Instead of detoxifying and assimilating ethanol like its Adh parental gene, we observe that JGW efficiently utilizes long-chain primary alcohols found in hormone and pheromone metabolism. These data suggest that protein functional diversity can expand rapidly under the joint forces of exon shuffling, gene duplication, and natural selection. PMID- 15534207 TI - Neurogenesis of corticospinal motor neurons extending spinal projections in adult mice. AB - The adult mammalian CNS shows a very limited capacity to regenerate after injury. However, endogenous precursors, or stem cells, provide a potential source of new neurons in the adult brain. Here, we induce the birth of new corticospinal motor neurons (CSMN), the CNS neurons that die in motor neuron degenerative diseases, including amyotrophic lateral sclerosis, and that cause loss of motor function in spinal cord injury. We induced synchronous apoptotic degeneration of CSMN and examined the fates of newborn cells arising from endogenous precursors, using markers for DNA replication, neuroblast migration, and progressive neuronal differentiation, combined with retrograde labeling from the spinal cord. We observed neuroblasts entering the neocortex and progressively differentiating into mature pyramidal neurons in cortical layer V. We found 20-30 new neurons per mm(3) in experimental mice vs. 0 in controls. A subset of these newborn neurons projected axons into the spinal cord and survived >56 weeks. These results demonstrate that endogenous precursors can differentiate into even highly complex long-projection CSMN in the adult mammalian brain and send new projections to spinal cord targets, suggesting that molecular manipulation of endogenous neural precursors in situ may offer future therapeutic possibilities for motor neuron degenerative disease and spinal cord injury. PMID- 15534208 TI - Integrated regulatory responses of fimB to N-acetylneuraminic (sialic) acid and GlcNAc in Escherichia coli K-12. AB - Bacterial-host attachment by means of bacterial adhesins is a key step in host colonization. Phase variation (reversible on-off switching) of the type 1 fimbrial adhesin of Escherichia coli involves a DNA inversion catalyzed by FimB (switching in either direction) or FimE (mainly on-to-off switching). fimB is separated from the divergent yjhATS operon by a large (1.4 kbp) intergenic region. Short ( approximately 28 bp) cis-active elements (regions 1 and 2) close to yjhA stimulate fimB expression and are required for sialic acid (Neu(5)Ac) sensitivity of its expression [El-Labany, S., Sohanpal, B. K., Lahooti, M., Akerman, R. & Blomfield, I. C. (2003) Mol. Microbiol. 49, 1109-1118]. Here, we show that whereas NanR, a sialic acid-response regulator, binds to region 1, NagC, a GlcNAc-6P-responsive protein, binds to region 2 instead. The NanR- and NagC-binding sites lie adjacent to deoxyadenosine methylase (Dam) methylation sites (5'-GATC) that are protected from modification, and the two regulators are shown to be required for methylation protection at regions 1 and 2, respectively. Mutations in nanR and nagC diminish fimB expression, and both fimB expression and FimB recombination are inhibited by GlcNAc (3- and >35-fold, respectively). Sialic acid catabolism generates GlcNAc-6-P, and whereas GlcNAc disrupts methylation protection by NagC alone, Neu(5)Ac inhibits the protection mediated by both NanR and NagC as expected. Type 1 fimbriae are proinflammatory, and host defenses enhance the release of both Neu(5)Ac and GlcNAc by a variety of mechanisms. Inhibition of type 1 fimbriation by these amino sugars may thus help balance the interaction between E. coli and its hosts. PMID- 15534209 TI - Resolution and evolution of the duck-billed platypus karyotype with an X1Y1X2Y2X3Y3X4Y4X5Y5 male sex chromosome constitution. AB - The platypus (2n = 52) has a complex karyotype that has been controversial over the last three decades. The presence of unpaired chromosomes and an unknown sex determining system especially has defied attempts at conventional analysis. This article reports on the preparation of chromosome-specific probes from flow-sorted chromosomes and their application in the identification and classification of all platypus chromosomes. This work reveals that the male karyotype has 21 pairs of chromosomes and 10 unpaired chromosomes (E1-E10), which are linked by short regions of homology to form a multivalent chain in meiosis. The female karyotype differs in that five of these unpaired elements (E1, E3, E5, E7, and E9) are each present in duplicate, whereas the remaining five unpaired elements (E2, E4, E6, E8, and E10) are absent. This finding indicates that sex is determined by the alternate segregation of the chain of 10 during spermatogenesis so that equal numbers of sperm bear either one of the two groups of five elements, i.e., five X and five Y chromosomes. Chromosome painting reveals that these X and Y chromosomes contain pairing (XY shared) and differential (X- or Y-specific) segments. Y differential regions must contain male-determining genes, and X differential regions should be dosage-compensated in the female. Two models for the evolution of the sex-determining system are presented. The resolution of the longstanding debate over the platypus karyotype is an important step toward the understanding of mechanisms of sex determination, dosage compensation, and karyotype evolution. PMID- 15534210 TI - A combinatorial model for the Macdonald polynomials. AB - We introduce a polynomial C(mu)[Z; q, t], depending on a set of variables Z = z(1), z(2),..., a partition mu, and two extra parameters q, t. The definition of C(mu) involves a pair of statistics (maj(sigma, mu), inv(sigma, mu)) on words sigma of positive integers, and the coefficients of the z(i) are manifestly in N[q,t]. We conjecture that C(mu)[Z; q, t] is none other than the modified Macdonald polynomial H(mu)[Z; q, t]. We further introduce a general family of polynomials F(T)[Z; q, S], where T is an arbitrary set of squares in the first quadrant of the xy plane, and S is an arbitrary subset of T. The coefficients of the F(T)[Z; q, S] are in N[q], and C(mu)[Z; q, t] is a sum of certain F(T)[Z; q, S] times nonnegative powers of t. We prove F(T)[Z; q, S] is symmetric in the z(i) and satisfies other properties consistent with the conjecture. We also show how the coefficient of a monomial in F(T)[Z; q, S] can be expressed recursively. maple calculations indicate the F(T)[Z; q, S] are Schur-positive, and we present a combinatorial conjecture for their Schur coefficients when the set T is a partition with at most three columns. PMID- 15534211 TI - A crucial arginine residue is required for a conformational switch in NifL to regulate nitrogen fixation in Azotobacter vinelandii. AB - NifL is an antiactivator that tightly regulates transcription of genes required for nitrogen fixation in Azotobacter vinelandii by controlling the activity of its partner protein NifA, a member of the family of sigma(54)-dependent transcriptional activators. Although the C-terminal region of A. vinelandii NifL shows homology to the transmitter domains of histidine protein kinases, signal transduction between NifL and NifA is conveyed by means of protein-protein interactions rather than by phosphorylation. Binding of the ligand 2-oxoglutarate to NifA plays a crucial role in preventing inhibition by NifL under conditions appropriate for nitrogen fixation. We have used a suppressor screen to identify a critical arginine residue (R306) in NifL that is required to release NifA from inhibition under appropriate environmental conditions. Amino acid substitutions at position 306 result in constitutive inhibition of NifA activity by NifL, thus preventing nitrogen fixation. Biochemical studies with one of the mutant proteins demonstrate that the substitution alters the conformation of NifL significantly and prevents the response of NifA to 2-oxoglutarate. We propose that arginine 306 is critical for the propagation of signals perceived by A. vinelandii NifL in response to the redox and fixed-nitrogen status and is required for a conformational switch that inactivates the inhibitory function of NifL under conditions appropriate for nitrogen fixation. PMID- 15534212 TI - Microarrays of small molecules embedded in biodegradable polymers for use in mammalian cell-based screens. AB - We developed a microarray-based system for screening small molecules in mammalian cells. This system is compatible with image-based screens and requires fewer than 100 cells per compound. Each compound is impregnated in a 200-microm-diameter disc composed of biodegradable poly-(D),(L)-lactide/glycolide copolymer. Cells are seeded on top of these discs, and compounds slowly diffuse out, affecting proximal cells. In contrast with microtiter-based screening, this system does not involve the use of wells or walls between each compound-treated group of cells. We demonstrate detection of the effects of a single compound in a large microarray, that diverse compounds can be released in this format, and that extended release over several days is feasible. We performed a small synthetic lethal screen and identified a compound (macbecin II) that has reduced activity in cells with RNA interference-mediated decrease in the expression of tuberous sclerosis 2. Thus, we have developed a microarray-based screening system for testing the effects of small molecules on mammalian cells by using an imaging based readout. This method will be useful to those performing small-molecule screens to discover new chemical tools and potential therapeutic agents. PMID- 15534214 TI - Associative learning shapes the neural code for stimulus magnitude in primary auditory cortex. AB - Since the dawn of experimental psychology, researchers have sought an understanding of the fundamental relationship between the amplitude of sensory stimuli and the magnitudes of their perceptual representations. Contemporary theories support the view that magnitude is encoded by a linear increase in firing rate established in the primary afferent pathways. In the present study, we have investigated sound intensity coding in the rat primary auditory cortex (AI) and describe its plasticity by following paired stimulus reinforcement and instrumental conditioning paradigms. In trained animals, population-response strengths in AI became more strongly nonlinear with increasing stimulus intensity. Individual AI responses became selective to more restricted ranges of sound intensities and, as a population, represented a broader range of preferred sound levels. These experiments demonstrate that the representation of stimulus magnitude can be powerfully reshaped by associative learning processes and suggest that the code for sound intensity within AI can be derived from intensity tuned neurons that change, rather than simply increase, their firing rates in proportion to increases in sound intensity. PMID- 15534213 TI - Remote hot spots mediate protein substrate recognition for the Cdc25 phosphatase. AB - Cdc25B is a phosphatase that catalyzes the dephosphorylation and activation of the cyclin-dependent kinases, thus driving cell cycle progression. We have identified two residues, R488 and Y497, located >20 A from the active site, that mediate protein substrate recognition without affecting activity toward small molecule substrates. Injection of Cdc25B wild-type but not the R488L or Y497A variants induces germinal vesicle breakdown and cyclin-dependent kinase activation in Xenopus oocytes. The conditional knockout of the cdc25 homolog (mih1) in Saccharomyces cerevisiae can be complemented by the wild type but not by the hot spot variants, indicating that protein substrate recognition by the Cdc25 phosphatases is an essential and evolutionarily conserved feature. PMID- 15534215 TI - Activation of Arp2/3 complex-dependent actin polymerization by plant proteins distantly related to Scar/WAVE. AB - The Arp2/3 complex, a highly conserved nucleator of F-actin polymerization, plays a key role in the regulation of actin dynamics eukaryotic cells. In animal cells and yeasts, Wiskott-Aldrich Syndrome protein (WASP)/suppressor of cAMP receptor (Scar)/WASP family verprolin homologous (WAVE) family proteins activate the Arp2/3 complex in response to localized cues. Like other eukaryotes, plants have an Arp2/3 complex, which has recently been shown to play an important role in F actin organization and cell morphogenesis. However, no activators of the Arp2/3 complex have been identified in plants, which lack obvious homologs of WASP/Scar/WAVE family proteins. Here, we identify a family of Scar/WAVE-related plant Arp2/3 activators. Like Scar/WAVE proteins, four proteins identified in Arabidopsis thaliana (AtSCAR1 to AtSCAR4) and one in maize (ZmSCAR1) have a C terminal WASP homology 2 (WH2)/acidic (WA)-verprolin homology/cofilin homology/acidic (VCA)-like domain, which we show can activate the bovine Arp2/3 complex. At their N termini, AtSCAR1 to ATSCAR4, along with a fifth protein lacking a VCA/WA-like domain at its C terminus (At4g18600), are related to the N terminal Scar homology domains of Scar/WAVE family proteins. Analysis of gene expression patterns suggests functional redundancy among members of the AtSCAR family. Full-length AtSCAR1 and ATSCAR3 proteins and their Scar homology domains bind in vitro to AtBRICK 1 (AtBRK1), the Arabidopsis homolog of HSPC300, a WAVE binding protein recently identified as a component of a complex implicated in the regulation of Scar/WAVE activity. Thus, AtSCAR proteins are likely to function in association with AtBRK1, and perhaps other Arabidopsis homologs of WAVE complex components, to regulate activation of the Arp2,3 complex in vivo. PMID- 15534217 TI - Molecular dynamics simulations of spontaneous fibril formation by random-coil peptides. AB - Assembly of normally soluble proteins into amyloid fibrils is a cause or associated symptom of numerous human disorders, including Alzheimer's and the prion diseases. We report molecular-level simulation of spontaneous fibril formation. Systems containing 12-96 model polyalanine peptides form fibrils at temperatures greater than a critical temperature that decreases with peptide concentration and exceeds the peptide's folding temperature, consistent with experimental findings. Formation of small amorphous aggregates precedes ordered nucleus formation and subsequent rapid fibril growth through addition of beta sheets laterally and monomeric peptides at fibril ends. The fibril's structure is similar to that observed experimentally. PMID- 15534216 TI - Proteins of purified Epstein-Barr virus. AB - Mature Epstein-Barr virus (EBV) was purified from the culture medium of infected lymphocytes made functionally conditional for Zta activation of lytic replication by an in-frame fusion with a mutant estrogen receptor. Proteins in purified virus preparations were separated by gradient gel electrophoresis and trypsin-digested; peptides were then analyzed by tandem hydrophobic chromatography, tandem MS sequencing, and MS scans. Potential peptides were matched with EBV and human gene ORFs. Mature EBV was mostly composed of homologues of proteins previously found in a herpes virion. However, EBV homologues to herpes simplex virus capsid associated or tegument components UL7 (BBRF2), UL14 (BGLF3), and EBV BFRF1 were not significantly detected. Instead, probable tegument components included the EBV and gamma-herpesvirus-encoded BLRF2, BRRF2, BDLF2 and BKRF4 proteins. Actin was also a major tegument protein, and cofilin, tubulin, heat shock protein 90, and heat shock protein 70 were substantial components. EBV envelope glycoprotein gp350 was highly abundant, followed by glycoprotein gH, intact and furin-cleaved gB, gM, gp42, gL, gp78, gp150, and gN. BILF1 (gp64) and proteins associated with latent EBV infection were not detected in virions. PMID- 15534219 TI - A versatile statistical analysis algorithm to detect genome copy number variation. AB - We have developed a versatile statistical analysis algorithm for the detection of genomic aberrations in human cancer cell lines. The algorithm analyzes genomic data obtained from a variety of array technologies, such as oligonucleotide array, bacterial artificial chromosome array, or array-based comparative genomic hybridization, that operate by hybridizing with genomic material obtained from cancer and normal cells and allow detection of regions of the genome with altered copy number. The number of probes (i.e., resolution), the amount of uncharacterized noise per probe, and the severity of chromosomal aberrations per chromosomal region may vary with the underlying technology, biological sample, and sample preparation. Constrained by these uncertainties, our algorithm aims at robustness by using a priorless maximum a posteriori estimator and at efficiency by a dynamic programming implementation. We illustrate these characteristics of our algorithm by applying it to data obtained from representational oligonucleotide microarray analysis and array-based comparative genomic hybridization technology as well as to synthetic data obtained from an artificial model whose properties can be varied computationally. The algorithm can combine data from multiple sources and thus facilitate the discovery of genes and markers important in cancer, as well as the discovery of loci important in inherited genetic disease. PMID- 15534218 TI - Thyroid hormone administration enhances remyelination in chronic demyelinating inflammatory disease. AB - Chronic disabilities in multiple sclerosis are believed to be due to neuron damage and degeneration, which follow remyelination failure. Due to the presence of numerous oligodendrocyte precursors inside demyelination plaques, one reason for demyelination failure could be the inability of oligodendrocyte precursor cells to turn into myelinating oligodendrocytes. In this study, we show that thyroid hormone enhances and accelerates remyelination in an experimental model of chronic demyelination, i.e., experimental allergic encephalomyelitis in congenic female Dark Agouti rats immunized with complete guinea pig spinal cord. Thyroid hormone, when administered during the acute phase of the disease, increases expression of platelet-derived growth factor alpha receptor, restores normal levels of myelin basic protein mRNA and protein, and allows an early and morphologically competent reassembly of myelin sheaths. Moreover, thyroid hormone exerts a neuroprotective effect with respect to axonal pathology. PMID- 15534220 TI - Higher-order assembly of microtubules by counterions: from hexagonal bundles to living necklaces. AB - Cellular factors tightly regulate the architecture of bundles of filamentous cytoskeletal proteins, giving rise to assemblies with distinct morphologies and physical properties, and a similar control of the supramolecular organization of nanotubes and nanorods in synthetic materials is highly desirable. However, it is unknown what principles determine how macromolecular interactions lead to assemblies with defined morphologies. In particular, electrostatic interactions between highly charged polyelectrolytes, which are ubiquitous in biological and synthetic self-assembled structures, are poorly understood. We have used a model system consisting of microtubules (MTs) and multivalent cations to examine how microscopic interactions can give rise to distinct bundle phases in biological polyelectrolytes. The structure of these supramolecular assemblies was elucidated on length scales from subnanometer to micrometer with synchrotron x-ray diffraction, transmission electron microscopy, and differential interference contrast microscopy. Tightly packed hexagonal bundles with controllable diameters were observed for large trivalent, tetravalent, and pentavalent counterions. Unexpectedly, in the presence of small divalent cations, we have discovered a living necklace bundle phase, comprised of 2D dynamic assemblies of MTs with linear, branched, and loop topologies. This new bundle phase is an experimental example of nematic membranes. The morphologically distinct MT assemblies give insight into general features of bundle formation and may be used as templates for miniaturized materials with applications in nanotechnology and biotechnology. PMID- 15534221 TI - Electron transfer between hemes in mammalian cytochrome c oxidase. AB - Fast intraprotein electron transfer reactions associated with enzymatic catalysis are often difficult to synchronize and therefore to monitor directly in non-light driven systems. However, in the mitochondrial respiratory enzyme cytochrome oxidase aa(3), the kinetics of the final electron transfer step into the active site can be determined: reverse electron flow between the close-lying and chemically identical hemes a(3) and a can be initiated by flash photolysis of CO from reduced heme a(3) under conditions where heme a is initially oxidized. To follow this reaction, we used transient absorption spectroscopy, with femtosecond time resolution and a time window extending to 4 ns. Comparison of the picosecond heme a(3)-CO photodissociation spectra under different redox states of heme a shows significant spectral interaction between both hemes, a phenomenon complicating the interpretation of spectral studies with low time resolution. Most importantly, we show that the intrinsic electron equilibration, corresponding to a DeltaG(0) of 45-55 meV, occurs in 1.2 +/- 0.1 ns. This is 3 orders of magnitude faster than the previously established equilibration phase of approximately 3 mus, which we suggest to reflect a change in redox equilibrium closely following CO migration out of the active site. Our results allow testing a number of conflicting predictions regarding this reaction between both experimental and theoretical studies. We discuss the potential physiological relevance of fast equilibration associated with this low-driving-force redox reaction. PMID- 15534222 TI - Interacting models of cooperative gene regulation. AB - Cooperativity between transcription factors is critical to gene regulation. Current computational methods do not take adequate account of this salient aspect. To address this issue, we present a computational method based on multivariate adaptive regression splines to correlate the occurrences of transcription factor binding motifs in the promoter DNA and their interactions to the logarithm of the ratio of gene expression levels. This allows us to discover both the individual motifs and synergistic pairs of motifs that are most likely to be functional, and enumerate their relative contributions at any arbitrary time point for which mRNA expression data are available. We present results of simulations and focus specifically on the yeast cell-cycle data. Inclusion of synergistic interactions can increase the prediction accuracy over linear regression to as much as 1.5- to 3.5-fold. Significant motifs and combinations of motifs are appropriately predicted at each stage of the cell cycle. We believe our multivariate adaptive regression splines-based approach will become more significant when applied to higher eukaryotes, especially mammals, where cooperative control of gene regulation is absolutely essential. PMID- 15534223 TI - Parametric inference for biological sequence analysis. AB - One of the major successes in computational biology has been the unification, by using the graphical model formalism, of a multitude of algorithms for annotating and comparing biological sequences. Graphical models that have been applied to these problems include hidden Markov models for annotation, tree models for phylogenetics, and pair hidden Markov models for alignment. A single algorithm, the sum-product algorithm, solves many of the inference problems that are associated with different statistical models. This article introduces the polytope propagation algorithm for computing the Newton polytope of an observation from a graphical model. This algorithm is a geometric version of the sum-product algorithm and is used to analyze the parametric behavior of maximum a posteriori inference calculations for graphical models. PMID- 15534224 TI - Tropical geometry of statistical models. AB - This article presents a unified mathematical framework for inference in graphical models, building on the observation that graphical models are algebraic varieties. From this geometric viewpoint, observations generated from a model are coordinates of a point in the variety, and the sum-product algorithm is an efficient tool for evaluating specific coordinates. Here, we address the question of how the solutions to various inference problems depend on the model parameters. The proposed answer is expressed in terms of tropical algebraic geometry. The Newton polytope of a statistical model plays a key role. Our results are applied to the hidden Markov model and the general Markov model on a binary tree. PMID- 15534225 TI - Groups of diverse problem solvers can outperform groups of high-ability problem solvers. AB - We introduce a general framework for modeling functionally diverse problem solving agents. In this framework, problem-solving agents possess representations of problems and algorithms that they use to locate solutions. We use this framework to establish a result relevant to group composition. We find that when selecting a problem-solving team from a diverse population of intelligent agents, a team of randomly selected agents outperforms a team comprised of the best performing agents. This result relies on the intuition that, as the initial pool of problem solvers becomes large, the best-performing agents necessarily become similar in the space of problem solvers. Their relatively greater ability is more than offset by their lack of problem-solving diversity. PMID- 15534226 TI - RGS9-2 modulates D2 dopamine receptor-mediated Ca2+ channel inhibition in rat striatal cholinergic interneurons. AB - Regulator of G protein signaling (RGS) proteins negatively regulate receptor mediated second messenger responses by enhancing the GTPase activity of Galpha subunits. We describe a receptor-specific role for an RGS protein at the level of an individual brain neuron. RGS9-2 and Gbeta(5) mRNA and protein complexes were detected in striatal cholinergic and gamma-aminobutyric acidergic neurons. Dialysis of cholinergic neurons with RGS9 constructs enhanced basal Ca(2+) channel currents and reduced D(2) dopamine receptor modulation of Cav2.2 channels. These constructs did not alter M(2) muscarinic receptor modulation of Cav2.2 currents in the same neuron. The noncatalytic DEP-GGL domain of RGS9 antagonized endogenous RGS9-2 activity, enhancing D(2) receptor modulation of Ca(2+) currents. In vitro, RGS9 constructs accelerated GTPase activity, in agreement with electrophysiological measurements, and did so more effectively at Go than Gi. These results implicate RGS9-2 as a specific regulator of dopamine receptor-mediated signaling in the striatum and identify a role for GAP activity modulation by the DEP-GGL domain. PMID- 15534227 TI - Induction of antiviral immunity requires Toll-like receptor signaling in both stromal and dendritic cell compartments. AB - Pattern recognition by Toll-like receptors (TLRs) is known to be important for the induction of dendritic cell (DC) maturation. DCs, in turn, are critically important in the initiation of T cell responses. However, most viruses do not infect DCs. This recognition system poses a biological problem in ensuring that most viral infections be detected by pattern recognition receptors. Furthermore, it is unknown what, if any, is the contribution of TLRs expressed by cells that are infected by a virus, versus TLRs expressed by DCs, in the initiation of antiviral adaptive immunity. Here we address these issues using a physiologically relevant model of mucosal infection with herpes simplex virus type 2. We demonstrate that innate immune recognition of viral infection occurs in two distinct stages, one at the level of the infected epithelial cells and the other at the level of the noninfected DCs. Importantly, both TLR-mediated recognition events are required for the induction of effector T cells. Our results demonstrate that virally infected tissues instruct DCs to initiate the appropriate class of effector T cell responses and reveal the critical importance of the stromal cells in detecting infectious agents through their own pattern recognition receptors. PMID- 15534228 TI - Guiding neuronal development with in situ microfabrication. AB - We report the ability to modify microscopic 3D topographies within dissociated cultures, providing a means to alter the development of neurons as they extend neurites and establish interconnections. In this approach, multiphoton excitation is used to focally excite noncytotoxic photosensitizers that promote protein crosslinking, such as BSA, into matrices having feature sizes >/=250 nm. Barriers, growth lanes, and pinning structures comprised of crosslinked proteins are fabricated under conditions that do not compromise the viability of neurons both on short time scales and over periods of days. In addition, the ability to fabricate functional microstructures from crosslinked avidin enables submicrometer localization of controllable quantities of biotinylated ligands, such as indicators and biological effectors. Feasibility is demonstrated for using in situ microfabrication to guide the contact position of cortical neurons with micrometer accuracy, opening the possibility for engineering well defined sets of synaptic interactions. PMID- 15534229 TI - Transgenic mice expressing a soluble form of porcine nectin-1/herpesvirus entry mediator C as a model for pseudorabies-resistant livestock. AB - An approach to genetically engineered resistance to pseudorabies virus (PRV) infection was examined by using a transgene encoding a soluble form of nectin-1, also known as herpesvirus entry mediator C. Nectin-1 is an alpha-herpesvirus receptor that binds to virion glycoprotein D. Nectin-1 mediates entry of PRV, herpes simplex virus types 1 and 2, and bovine herpesvirus type 1. To assess the antiviral potential of an ectopic expression of the nectin-1 ectodomain in vivo, six transgenic mouse lines expressing a soluble form of nectin-1, consisting of an extracellular domain of porcine nectin-1 and the Fc portion of human IgG1, were generated. All of the transgenic mouse lines showed nearly complete resistance to PRV infection by means of both i.p. and intranasal routes. These results suggest that the introduction into farm animals of a transgene encoding a soluble form of nectin-1 would offer a potent biological approach to generating alpha-herpesvirus-resistant livestock. PMID- 15534230 TI - Selenophosphate synthetase genes from lung adenocarcinoma cells: Sps1 for recycling L-selenocysteine and Sps2 for selenite assimilation. AB - A labile selenium donor compound monoselenophosphate is synthesized from selenide and ATP by selenophosphate synthetase (SPS). In the present study, Sps1 and Sps2 were cloned from a cDNA library prepared from human lung adenocarcinoma cells (NCIH441). The human lung Sps1 has been cloned as an ORF of 1,179 bp, identical in sequence to that of the recently revised human liver Sps1. The in-frame TGA codon of the lung Sps2 was genetically altered to TGT (Cys) to obtain the Sps2Cys gene. Expression of the recombinant plasmids containing Sps1 or Sps2Cys was highly toxic to Escherichia coli host cells grown aerobically. Accordingly, the human lung Sps homologs were characterized by an in vivo complementation assay using a selD mutant strain. An added selenium source and a low salt concentration (0.1-0.25% NaCl) in the medium were required for reproducible and sensitive in vivo complementation. Sps2Cys effectively complemented the selD mutant, and the resulting formate dehydrogenase H activity was as high as that of WT E. coli MC4100. In contrast, only a weak complementation of the selD mutant by the Sps1 gene was observed when cells were grown in selenite media. Better complementation with added l-selenocysteine suggested involvement of a selenocysteine lyase for mobilization of selenium. Based on this apparent substrate specificity of the Sps1 and Sps2 gene products we suggest that the Sps1-encoded enzyme depends on a selenium salvage system that recycles l-selenocysteine, whereas the Sps2 enzyme can function with a selenite assimilation system. PMID- 15534231 TI - Patient page. Attacks of immobility caused by diet or exercise? The mystery of periodic paralyses. PMID- 15534232 TI - Treating cognitive deficits in multiple sclerosis: are we there yet? PMID- 15534233 TI - Antisaccades: probing cognitive flexibility with eye movements. PMID- 15534234 TI - American Academy of Neurology policy on expert medical testimony. PMID- 15534235 TI - Expert medical testimony: Responsibilities of medical societies. AB - Expert witnesses for plaintiff and defense should be able to provide evidence based literature to support their testimony in depositions and in the courtroom. Specialty societies, such as the American Academy of Neurology, should be willing to review complaints about unsubstantiated testimony and, at times, publicly discipline offending experts. PMID- 15534236 TI - Neurocysticercosis in the United States: review of an important emerging infection. AB - OBJECTIVE: To review published clinical studies on neurocysticercosis (NCC) in the United States over the past two decades and comment on epidemiologic trends and treatment. METHODS: This review is based on a search of the literature citing NCC cases diagnosed in the United States utilizing PUBMED for the years 1980 through early 2004. Case series, case reports, epidemiologic studies, and treatment of NCC were evaluated. RESULTS: A total of 1,494 patients with NCC were reported in the United States among large case series (n > 20) between 1980 and early 2004. Common onset symptoms for these patients included seizures (66%), hydrocephalus (16%), and headaches (15%). The majority presented with parenchymal NCC (91%), with the remainder having ventricular cysts (6%), subarachnoid cysts (2%), and spinal cysts (0.2%). A total of 76 cases of NCC were likely acquired within the United States during the period of this review. A higher risk for acquiring NCC has been documented in patients who have traveled to endemic regions, are of Hispanic ethnicity, and have contact with Taenia solium tapeworm carriers. CONCLUSIONS: An increasing number of NCC cases have been reported in the US literature over the past 50 years, suggesting that the prevalence of this disease may be on the rise. Because neurologists are often involved with the diagnosis and management of NCC in the United States, it is important that they become familiar with this disorder, as they will play an important role in efforts to control the disease. PMID- 15534237 TI - Injuries due to seizures in persons with epilepsy: a population-based study. AB - BACKGROUND: Previous studies of injuries due to epileptic seizures predominantly involved patients with intractable epilepsy. These studies may have overestimated the risk of injuries in persons with epilepsy. METHODS: Patients consisted of 247 Rochester, MN, residents who were diagnosed with epilepsy between 1975 and 1984. Seizure-related injuries were defined as any injury, other than orolingual trauma, resulting from a seizure, sufficient for the patient to seek medical attention or for injury occurrence to be determined during the course of medical care. To identify risk factors for injury, characteristics of patients with seizure-related injury were compared with those without injury. RESULTS: During a total of 2,714 patient-years of follow-up, 62 seizure-related injuries were identified in 39 patients (16%, one injury in every 44 person-years). Most injuries involved cranial soft tissue contusions or lacerations (79%). The majority of seizure-related injuries (82%) occurred during generalized convulsive seizures. Univariate analyses identified five potential risk factors for seizure related injury: greater number of antiepileptic drugs used, less independent living situation, higher Rankin score, history of generalized convulsive seizures or drop attacks, and higher seizure frequency score. Seizure frequency, however, was the only significant risk factor identified by multivariate analysis (p < 0.001; relative risk, 1.33). CONCLUSIONS: This population-based study shows that seizure-related injuries are infrequent and generally of minor severity. In most epilepsy patients, excessive restriction of daily activities to avoid injury is unnecessary. Effective seizure control reliably reduces the risk of seizure related injuries. PMID- 15534238 TI - Neural substrate of antisaccades: role of subcortical structures. AB - BACKGROUND: Experimental and clinical studies suggest that the dorsolateral prefrontal cortex (DLPFC) and the superior colliculus (SC) are crucial for the cancellation of reflexive eye movements toward distracting stimuli. However, the contribution of subcortical structures remains unknown. The basal ganglia provide serial tonic inhibitory connections between the DLPFC and the SC, and could therefore be involved in preventing the triggering of unnecessary saccades. The DLPFC could also exert its inhibitory effect on the SC through direct prefronto tectal pathways that travel in the internal capsule (IC). Since thalamic dysfunction may be responsible for reduced DLPFC activation, it may be hypothesized that the thalamus could also participate in saccadic inhibition. METHODS: The authors recorded reflexive saccade triggering (prosaccade task) and inhibition (antisaccade task) in 29 patients with a single lesion affecting the striatum, the thalamus, or the IC, and compared these results to control subjects. RESULTS: A normal error rate in the antisaccade task was found in patients with 1) a basal ganglia lesion, 2) a thalamic lesion, or 3) a lesion restricted to the posterior half of the posterior limb of the IC. An increased error rate in the antisaccade task was found in patients with a lesion affecting the anterior limb, the genu, or the anterior half of the posterior limb of the IC. CONCLUSION: These results suggest that neither the basal ganglia nor the thalamus plays a major role in reflexive saccade suppression, but support the hypothesis of a direct DLPFC inhibitory control of saccade triggering on the SC. PMID- 15534239 TI - Donepezil improved memory in multiple sclerosis in a randomized clinical trial. AB - OBJECTIVE: To determine the effect of donepezil in treating memory and cognitive dysfunction in multiple sclerosis (MS). METHODS: This single-center double-blind placebo-controlled clinical trial evaluated 69 MS patients with cognitive impairment who were randomly assigned to receive a 24-week treatment course of either donepezil (10 mg daily) or placebo. Patients underwent neuropsychological assessment at baseline and after 24 weeks of treatment. The primary outcome was change in verbal learning and memory on the Selective Reminding Test (SRT). Secondary outcomes included other tests of cognitive function, patient-reported change in memory, and clinician-reported impression of cognitive change. RESULTS: Donepezil-treated patients showed significant improvement in memory performance on the SRT compared to placebo (p = 0.043). The benefit of donepezil remained significant after controlling for various covariates including age, Expanded Disability Status Scale, baseline SRT score, reading ability, MS subtype, and sex. Donepezil-treated patients did not show significant improvements on other cognitive tests, but were more than twice as likely to report memory improvement than those in the placebo group (p = 0.006). The clinician also reported cognitive improvement in almost twice as many donepezil vs placebo patients (p = 0.036). No serious adverse events related to study medication occurred, although more donepezil (34.3%) than placebo (8.8%) subjects reported unusual/abnormal dreams (p = 0.010). CONCLUSIONS: Donepezil improved memory in MS patients with initial cognitive impairment in a single center clinical trial. A larger multicenter investigation of donepezil in MS is warranted in order to more definitively assess the efficacy of this intervention. PMID- 15534240 TI - Neurofilament and glial fibrillary acidic protein in multiple sclerosis. AB - OBJECTIVE: To evaluate levels of neurofilament light (NFL) and glial fibrillary acidic protein (GFAP) in CSF from patients with multiple sclerosis (MS) in relation to clinical progress of the disease. METHODS: CSF levels of NFL and GFAP were determined by sensitive ELISAs in 99 patients with different subtypes of MS, classified in terms of "ongoing relapse" or "clinically stable disease," and 25 control subjects. Levels were compared with paraclinical data such as immunoglobulin G index and inflammatory cell count in the CSF, and the levels were related to Expanded Disability Status Scale score and progression index at clinical follow-up evaluations later in the disease course. RESULTS: NFL and GFAP levels were elevated in MS patients as compared with control subjects (p < 0.001). The NFL levels were higher at relapses, whereas GFAP levels were unaffected. High NFL levels correlated with progression in patients with an active relapse (r = 0.49; p < 0.01) and in clinically stable patients (r = 0.29; p < 0.05). GFAP correlated to progression in the total patient cohort (r = 0.24; p < 0.05). Moreover, a strong correlation between NFL levels and inflammatory cell counts was evident in the group of patients with an ongoing relapse (r = 0.52; p = 0.001). CONCLUSIONS: CSF levels of neurofilament light and glial fibrillary acidic protein may have prognostic value in multiple sclerosis. PMID- 15534241 TI - Stroke risk profile, brain volume, and cognitive function: the Framingham Offspring Study. AB - BACKGROUND: Mid-life stroke risk factors have been related to late-life cognitive impairment. This association may result not only from clinical strokes but also from subclinical brain injury, such as a global atrophy demonstrable on quantitative brain MRI. METHODS: The authors evaluated the community-based cohort of Framingham Offspring Study participants. A total of 1,841 subjects (mean age, 62 years; 857 men, 984 women) who underwent quantitative MRI and cognitive testing between 1999 and 2001 and were free of clinical stroke and dementia constituted our study sample. The authors used age- and sex-adjusted linear regression models to relate previous (1991 to 1995) and recent (1998 to 2001) Framingham Stroke Risk Profile (FSRP) scores to the total cerebral brain volume ratio (TCBVr) on follow-up MRI, and further to relate the TCBVr with education adjusted scores on neuropsychological tests administered at the time of imaging. RESULTS: There was an inverse association between FSRP scores and TCBVr. The TCBVr also showed a significant positive association with performance on tests of attention (Trails A), executive function (Trails B), and visuospatial function (visual reproduction, Hooper visual organization), but not with performance on tests of verbal memory or naming. CONCLUSIONS: The Framingham Stroke Risk Profile may identify subjects with smaller brains and poorer cognitive function among stroke- and dementia-free subjects, reinforcing the importance of managing stroke risk factors. PMID- 15534242 TI - Right spatial neglect after left hemisphere stroke: qualitative and quantitative study. AB - OBJECTIVES: Comparatively little research has been conducted on right neglect after left brain damage. The authors sought to assess contralateral neglect in subacute left hemisphere stroke patients using a comprehensive test battery validated in a large control group after right hemisphere stroke. METHODS: Seventy-eight left hemisphere stroke patients were assessed. The test battery included a preliminary assessment of anosognosia and visual extinction, a clinical assessment of gaze orientation and personal neglect, and paper-and pencil tests of spatial neglect in the peripersonal space. Only nonverbal tests were used. RESULTS: Drawing and cancellation tasks revealed neglect in 10 to 13% of patients. The combined battery was more sensitive than any single test alone. A total of 43.5% of patients showed some degree of neglect on at least one measure. Anatomic analyses showed that neglect was more common and severe when the posterior association cortex was damaged. CONCLUSIONS: The frequency of occurrence of right neglect was, as expected, much lower than that reported in a study using the same assessment battery in right brain damage stroke patients. Nevertheless, neglect was found in a substantial proportion of patients at a subacute stage, suggesting that it should be considered in the rehabilitation planning of left brain damage stroke patients. PMID- 15534243 TI - White matter lesions, cognition, and recurrent hemorrhage in lobar intracerebral hemorrhage. AB - BACKGROUND: Accumulating evidence suggests that white matter lesions are associated with vascular cognitive impairment. The authors investigated the relationships between white matter lesions, cognitive impairment, and risk of recurrent hemorrhage in a prospectively identified cohort of patients with lobar intracerebral hemorrhage (ICH). METHODS: The authors collected clinical and genetic information on 182 consecutive patients age > or = 55 who had CT scan at admission for lobar ICH. White matter disease was graded on CT in all subjects and on MRI in a subset of 82 patients. All scans were interpreted blinded to clinical information. Survivors were followed for recurrent ICH by telephone interview. RESULTS: White matter damage was common (present on CT in 77%) and severe (advanced CT grade in 32%). White matter damage was correlated with the total number of hemorrhages on gradient-echo MRI and with risk of recurrent ICH. Subjects with cognitive impairment prior to their index ICH were more likely to have severe white matter damage on CT (OR 3.6, 95% CI 1.6 to 8.1, p = 0.003) and more likely to have advanced periventricular hyperintensities on MRI. The relationships between white matter damage and cognitive impairment were similar in the subset of 88 subjects meeting criteria for probable or definite cerebral amyloid angiopathy and remained independent after adjustment for age, cortical atrophy, and APOE genotype. CONCLUSIONS: White matter damage in lobar ICH is common and is associated with cognitive impairment. These data support the possibility that an underlying vasculopathy in lobar ICH patients, possibly cerebral amyloid angiopathy, can cause clinically important vascular dysfunction. PMID- 15534244 TI - Hemorrhage is uncommon in new Alzheimer family with Flemish amyloid precursor protein mutation. AB - BACKGROUND: Most mutations in the amyloid precursor protein (APP) gene have been associated with familial Alzheimer disease (AD); however, some mutations within the Abeta-coding sequence have been described in families with recurrent cerebral hemorrhage. The APPAla692Gly (Flemish) mutation was reported in a family in which affected members developed hemorrhagic stroke, progressive dementia, or both. OBJECTIVE: To describe clinical, neuropathologic, and genetic features of a family of British origin with the Flemish APP mutation. METHODS: Clinical features of the proband and two affected relatives were obtained by history, examination, and medical record review. Some information on deceased affected relatives was obtained by informant interview. Neuropathologic examination was carried out on one case. DNA studies were carried out on three affected and three unaffected individuals. RESULTS: Presenile dementia was present in a pattern consistent with dominant inheritance, with the APP692 mutation being found in all affecteds and no unaffecteds. The proband also had a cerebral hemorrhage, but was the only one of five affecteds to have this complication. Neuropathologic examination confirmed AD, congophilic angiopathy, and hemorrhagic infarction. CONCLUSIONS: This expands the number of families reported with mutations in the coding region of the amyloid precursor protein gene. Cerebral hemorrhage appears to be less frequent in this family than in the previously reported Flemish pedigree with the same mutation. PMID- 15534245 TI - Progression of cognitive impairment in stroke patients. AB - OBJECTIVE: To examine the progression of neuropsychological deficits in stroke patients with and without cognitive impairment. METHODS: The authors assessed the Sydney Stroke Study cohort 1 year after index assessment with detailed neuropsychological and medical-psychiatric assessments. The neuropsychological tests were classified into cognitive domains, and composite z-scores adjusted for age and education. Changes in cognitive test scores were compared between groups and predictors of cognitive change examined. RESULTS: Patients (n = 128) had a mean decline of 0.83 (SD 2.2) points on the Mini-Mental State Examination (MMSE) compared to an increase of 0.76 (1.3) in controls (n = 78) (p < 0.0001), and a small but significant decline in informant ratings of function and cognition. The decline on a composite index of cognitive function was not significantly different in the groups after correction for age, education, and index assessment cognitive function. Stroke/transient ischemic attack patients, however, had greater decline in verbal memory and visuoconstructive function. The occurrence of an interval stroke (n = 14) significantly increased the cognitive decline to a mean 2.0 points on the MMSE. The rate of change had a significant correlation (r = 0.24) with white matter hyperintensity volume at index assessment. On regression analysis the only predictor of cognitive change was years of education, which had a protective function. CONCLUSIONS: Subjects with cerebrovascular disease have a slow decline in cognitive functioning in the absence of further cerebrovascular events, although the occurrence of such an event accentuates the dysfunction. Education plays a protective role. PMID- 15534246 TI - Statin therapy and risk of dementia in the elderly: a community-based prospective cohort study. AB - OBJECTIVE: To assess the association between statin therapy and risk of Alzheimer disease (AD) in a prospective cohort study with documented statin exposure and incident dementia. METHODS: This is a prospective, cohort study of statin use and incident dementia and probable AD. A cohort of 2,356 cognitively intact persons, aged 65 and older, were randomly selected from a health maintenance organization (HMO), and were assessed biennially for dementia. Statin use was identified using the HMO pharmacy database. A proportional hazards model with statin use as a time dependent covariate was used to assess the statin-dementia/AD association. RESULTS: Among 312 participants with incident dementia, 168 had probable AD. The unadjusted hazard ratios (HRs) with statin use were 1.33 (95% CI 0.95 to 1.85) for all-cause dementia and 0.90 (CI 0.54 to 1.51) for probable AD. Adjusted corresponding HRs were 1.19 (CI 0.82 to 1.75) and 0.82 (CI 0.46 to 1.46). A subgroup analysis of participants with at least one APOE-epsilon4 allele who entered the study before age 80 produced an adjusted HR of 0.33 (CI 0.10 to 1.04). CONCLUSION: Employing time-dependent proportional hazards modeling, the authors found no significant association between statin use and incident dementia or probable AD. In contrast, when the data were analyzed, inappropriately, as a case-control study, the authors found an OR of 0.55 for probable AD, falsely indicating a protective effect of statins. Study design and analytic methods may explain the discrepancy between the current null findings and earlier findings. PMID- 15534247 TI - Capturing the true burden of dystonia on patients: the Cervical Dystonia Impact Profile (CDIP-58). AB - OBJECTIVES: To develop a new rating scale for measuring the health impact of cervical dystonia (CD) that includes patients' perceptions and complements existing observer dependent clinician rating scales. METHODS: Scale development was in three stages. In Stage 1, a large pool of items was generated from patient interviews (n = 25), expert opinion, and literature review. In Stage 2, these items were administered by postal survey to people with CD. The resulting data were analyzed using Rasch item analysis to construct, from the item pool, a rating scale that satisfied criteria for rigorous measurement. In Stage 3, the measurement properties of this rating scale were examined in an independent sample of people with CD. RESULTS: In Stage 1, 150 items concerning the health impact of CD were generated. In Stage 2, 556 people completed questionnaires (87% response rate) and a 58-item rating scale measuring the health impact of CD in eight areas was constructed (CD Impact Profile, CDIP-58). In Stage 3, CDIP-58 data from 391 people (87% response rate) were received. Analyses supported the measurement of eight unidimensional constructs (infit mean square range 0.62 to 1.50), item calibration (33.37 to 67.56), and patient separation statistics (2.59 to 3.38). Items demonstrated stable calibrations in subgroups of people with CD supporting the stability of the CDIP-58. CONCLUSIONS: The CDIP-58 is a reliable and valid patient-based rating scale measuring the health impact of CD in eight health dimensions. PMID- 15534248 TI - Integrative cortical dysfunction and pervasive motion perception deficit in fragile X syndrome. AB - BACKGROUND: Fragile X syndrome (FXS) is associated with neurologic deficits recently attributed to the magnocellular pathway of the lateral geniculate nucleus. OBJECTIVE: To test the hypotheses that FXS individuals 1) have a pervasive visual motion perception impairment affecting neocortical circuits in the parietal lobe and 2) have deficits in integrative neocortical mechanisms necessary for perception of complex stimuli. METHODS: Psychophysical tests of visual motion and form perception defined by either first-order (luminance) or second-order (texture) attributes were used to probe early and later occipito temporal and occipito-parietal functioning. RESULTS: When compared to developmental- and age-matched controls, FXS individuals displayed severe impairments in first- and second-order motion perception. This deficit was accompanied by near normal perception for first-order form stimuli but not second order form stimuli. CONCLUSIONS: Impaired visual motion processing for first- and second-order stimuli suggests that both early- and later-level neurologic function of the parietal lobe are affected in Fragile X syndrome (FXS). Furthermore, this deficit likely stems from abnormal input from the magnocellular compartment of the lateral geniculate nucleus. Impaired visual form and motion processing for complex visual stimuli with normal processing for simple (i.e., first-order) form stimuli suggests that FXS individuals have normal early form processing accompanied by a generalized impairment in neurologic mechanisms necessary for integrating all early visual input. PMID- 15534249 TI - Aberrant auditory processing and atypical planum temporale in developmental stuttering. AB - OBJECTIVE: To learn if people with persistent developmental stuttering and atypical anatomy of their auditory temporal cortex have, when compared to control subjects, changes in fluency induced with delayed auditory feedback (DAF). BACKGROUND: DAF improves fluency in many individuals who stutter, and induces dysfluency in some normal people. The planum temporale (PT), a portion of auditory temporal cortex, is anatomically atypical in some adults who stutter and atypical anatomy might induce aberrant function. Thus, the people who demonstrate the paradoxical response to DAF might be those who have atypical anatomy. METHODS: Experimental subjects were adults with developmental stuttering (n = 14) and control subjects (n = 14) matched for age, sex, education, and handedness. Volumetric MRI scans of all subjects were obtained and the PT was measured in the right and left hemispheres. Based on these scans, subjects were classified as typical (leftward PT asymmetry) or atypical (rightward PT asymmetry). Prose passages were read at baseline, with non-altered feedback (NAF), and with DAF, and fluency was measured in these three conditions. RESULTS: At baseline the adults with developmental stuttering were significantly more dysfluent than controls (p < 0.0005). Controls' fluency did not significantly change with DAF, but DAF improved fluency in adults with developmental stuttering (p < 0.0005). In the stutter group enhanced fluency was associated with atypical (rightward) PT asymmetry, and the presence of typical (leftward) PT asymmetry was not associated with any significant change in fluency. The individuals with atypical PT asymmetry also had more severe stuttering at baseline compared to the experimental subjects with typical PT anatomy. CONCLUSIONS: In adults with persistent developmental stuttering and atypical PT anatomy, fluency is improved with DAF. These experimental subjects who showed improvement had more severe stuttering at baseline. Anomalous PT anatomy may be a neural risk for developmental stuttering in some individuals. Although a number of explanations are tenable, it may be that atypical rightward PT asymmetry may alter speech feedback, and treatment with DAF might allow these people to compensate. PMID- 15534250 TI - Correlating phenotype and genotype in the periodic paralyses. AB - BACKGROUND: Periodic paralyses and paramyotonia congenita are rare disorders causing disabling weakness and myotonia. Mutations in sodium, calcium, and potassium channels have been recognized as causing disease. OBJECTIVE: To analyze the clinical phenotype of patients with and without discernible genotype and to identify other mutations in ion channel genes associated with disease. METHODS: The authors have reviewed clinical data in patients with a diagnosis of hypokalemic periodic paralysis (56 kindreds, 71 patients), hyperkalemic periodic paralysis (47 kindreds, 99 patients), and paramyotonia congenita (24 kindreds, 56 patients). For those patients without one of the classically known mutations, the authors analyzed the entire coding region of the SCN4A, KCNE3, and KCNJ2 genes and portions of the coding region of the CACNA1S gene in order to identify new mutations. RESULTS: Mutations were identified in approximately two thirds of kindreds with periodic paralysis or paramyotonia congenita. The authors found differences between the disorders and between those with and without identified mutations in terms of age at onset, frequency of attacks, duration of attacks, fixed proximal weakness, precipitants of attacks, myotonia, electrophysiologic studies, serum potassium levels, muscle biopsy, response to potassium administration, and response to treatment with acetazolamide. CONCLUSIONS: Hypokalemic periodic paralysis, hyperkalemic periodic paralysis, and paramyotonia congenita may be distinguished based on clinical data. This series of 226 patients (127 kindreds) confirms some clinical features of this disorder with notable exceptions: In this series, patients without mutations had a less typical clinical presentation including an older age at onset, no changes in diet as a precipitant, and absence of vacuolar myopathy on muscle biopsy. PMID- 15534252 TI - Chronic immune sensory polyradiculopathy: a possibly treatable sensory ataxia. AB - BACKGROUND: Chronic inflammatory neuropathies can present with a sensory ataxia due to involvement of dorsal root ganglia (DRG) or sensory nerves. Selective inflammatory involvement of sensory nerve roots proximal to the DRG has been postulated. METHODS: The authors identified 15 patients with a sensory syndrome and normal nerve conduction studies. Sensory nerve root involvement was suggested by either somatosensory evoked potential (SSEP) or imaging abnormalities. CNS disease was excluded. RESULTS: All patients had gait ataxia, large fiber sensory loss, and paresthesias, and nine had frequent falls. The disease course was chronic and progressive (median duration 5 years, range 3 months to 18 years). Sural sensory nerve action potential amplitudes were preserved and SSEP abnormalities were consistent with sensory nerve root involvement. Five patients had enlargement of lumbar nerve roots on MRI with enhancement in three. The CSF protein was elevated in 13 of 14 patients tested. Three patients had lumbar sensory rootlet biopsies that showed thickened rootlets, decreased density of large myelinated fibers, segmental demyelination, onion-bulb formation, and endoneurial inflammation. Six patients who required aids to walk were treated with immune modulating therapy and all had marked improvement with four returning to normal ambulation. CONCLUSION: Based on the described clinical features, normal nerve conduction studies, characteristic somatosensory evoked potential (SSEP) abnormality, enlarged nerve roots, elevated CSF protein, and inflammatory hypertrophic changes of sensory nerve rootlet tissue, we suggest the term chronic immune sensory polyradiculopathy (CISP) for this syndrome. This condition preferentially affects large myelinated fibers of the posterior roots, may respond favorably to treatment, and may be a restricted form of chronic inflammatory demyelinating polyradiculoneuropathy. PMID- 15534251 TI - A clinical trial of creatine in ALS. AB - BACKGROUND: Mitochondrial dysfunction occurs early in the course of ALS, and the mitochondria may be an important site for therapeutic intervention. Creatine stabilizes the mitochondrial transition pore, and is important in mitochondrial ATP production. In a transgenic mouse model of ALS, administration of creatine prolongs survival and preserves motor function and motor neurons. METHODS: The authors conducted a randomized double-blind, placebo controlled trial on 104 patients with ALS from 14 sites to evaluate the efficacy of creatine supplementation in ALS. The primary outcome measure was maximum voluntary isometric contraction of eight upper extremity muscles, with secondary outcomes including grip strength, ALS Functional Rating Scale-Revised, and motor unit number estimates. Patients were treated for 6 months, and evaluated monthly. RESULTS: Creatine was tolerated well, but no benefit of creatine could be demonstrated in any outcome measure. CI analysis showed that the study, although powered to detect a 50% or greater change in rate of decline of muscle strength, actually made an effect size of greater than 23% unlikely. It was also demonstrated that motor unit number estimation was performed with acceptable reproducibility and tolerability, and may be a useful outcome measure in future clinical trials. CONCLUSION: Any beneficial effect of creatine at 5 g per day in ALS must be small. Other agents should be considered in future studies of therapeutic agents to address mitochondrial dysfunction in ALS. In addition, motor unit number estimation may be a useful outcome measure for future clinical trials in ALS. PMID- 15534253 TI - Decrement in surface-recorded motor unit potentials in amyotrophic lateral sclerosis. AB - BACKGROUND: A decrement to repetitive stimulation sometimes occurs in ALS. The ability to isolate individual surface-recorded motor unit potentials (SMUPs) indirectly allows the analysis of the decrement by comparing the effect of different rates of stimulation on the size of the isolated SMUP. METHODS: The median or ulnar nerve or both were studied in 11 patients with reduced compound muscle action potential (CMAP) amplitude. Isolated SMUPs were identified and studied using standard electrophysiologic techniques. One- and 3-Hz stimulation frequencies were compared in isolated SMUP by collecting > or =100 responses to a nonvarying stimulus. Changes in individual SMUP size and activation with change in stimulus frequency were compared with the size of the SMUP and with the CMAP decrement to a train-of-four stimulus. RESULTS: Fifteen studies were obtained from the 11 subjects. Eight studies showed a decrement of the total CMAP to a train-of-four stimulus. Twenty-nine well-defined, isolated SMUPs were studied. SMUP size reductions of 1 to 16% occurred with an increase in the stimulus frequency. No conduction block of individual SMUP occurred with the SMUP size reduction. No consistent change in variance was found. The SMUP size reduction was related to the size of the CMAP and the CMAP train-of-four decrement and less so to the size of the SMUP. CONCLUSION: The decrement of the total compound muscle action potential to repetitive stimulation appears to arise from individual surface-recorded motor unit potentials and does not result from loss of activation of the main motor axon. The probable mechanism lies at the nerve terminal and neuromuscular junction. No consistent change in variance occurs with different rates of stimulation. PMID- 15534254 TI - Ulnar neuropathy at the elbow: follow-up and prognostic factors determining outcome. AB - OBJECTIVE: To determine the outcome in patients with ulnar neuropathy at the elbow (UNE) treated surgically or conservatively, and the prognostic value of clinical, sonographic, and electrophysiologic features. METHODS: After a median follow-up of 14 months, 69 of 84 patients initially included in a prospective blinded study on the diagnostic value of sonography in UNE were re-evaluated. The patients underwent renewed systematic clinical and sonographic examination. Patients were scored as having a poor (stable or progressive symptoms) or favorable (complete remission of symptoms or improvement) outcome. RESULTS: Of the 74 initially affected arms, 12 (16%) had a complete remission, 21 (28%) improved, 25 (34%) remained stable, and 16 (22%) had progression. Surgically treated patients (28 arms) had a more favorable outcome than those treated conservatively (p = 0.03). After surgery, the mean ulnar nerve diameter decreased from 3.2 to 2.9 mm (p = 0.03), while this was not seen after conservative treatment. Multiple logistic regression analysis showed that more outspoken nerve enlargement found during sonography at the time of the diagnosis was associated with a poor outcome (OR: 2.9, p = 0.009). Furthermore, the presence of a motor conduction block (OR: 0.2, p = 0.03) and motor velocity slowing across the elbow (OR: 0.1, p = 0.01) were associated with a favorable outcome. CONCLUSION: More pronounced ulnar nerve thickening at the time of the diagnosis is associated with poor outcome at follow-up, especially in conservatively treated cases, while electrodiagnostic signs of demyelination on testing indicate favorable outcome. PMID- 15534255 TI - Evaluation of neuromuscular symptoms in UK Gulf War veterans: a controlled study. AB - OBJECTIVES: To determine whether Gulf War veterans with neuromuscular symptoms that included weakness and fatigue had either 1) objective correlates for muscle weakness or fatigue; or 2) any etiologic explanation for such symptoms; and if so, 3) whether such objective measures or etiologic mechanisms were specific to Gulf War service. METHODS: Forty-nine ill Gulf War veterans with more than four neuromuscular symptoms (Gulf-ill) were compared with 26 Gulf-well veterans, 13 symptomatic Bosnian veterans (Bosnia-ill), and 22 symptomatic troops who were not deployed to the Gulf (Era-ill). Quantitative myometry was used to objectively measure weakness and fatigue. Subjects had an ischemic forearm exercise test, a subanaerobic bicycle exercise test, and a muscle biopsy. RESULTS: Quantitative strength and fatigue measures did not correlate with self-perception of weakness or fatigue for any of our groups. No specific muscle biopsy abnormalities were found. There was no defect of adenylate deaminase or glycogenolysis found. Gulf ill subjects did find the subanaerobic bicycle exercise more effortful and generated significantly higher plasma lactate concentrations compared with Gulf well subjects. CONCLUSION: Because complaints of weakness and fatigue in unwell servicemen do not correlate with actual weakness or fatigue, explanations for these symptoms must lie outside of the neuromuscular system. Increased lactate production during subanaerobic bicycle exercise reflects mitochondrial inefficiency, but it is unclear whether this reflects mitochondrial damage sustained during Gulf War service or inactivity secondary to ill health. PMID- 15534256 TI - Late-onset Pompe disease primarily affects quality of life in physical health domains. AB - OBJECTIVE: To investigate quality of life in an international population of patients with late-onset Pompe disease. METHODS: Data on quality of life (SF-36), age, sex, disease duration, wheelchair use, and use of artificial ventilation were collected for 210 adults with Pompe disease from Australia, Germany, the Netherlands, the United Kingdom, and the United States. SF-36 scores were compared between countries and related to patient characteristics. In addition, for the Dutch subgroup (n = 51), comparisons with the general population and 1 year follow-up assessments were performed. RESULTS: No significant differences between countries were found for the four physical health scales. Mean scores on the vitality, role functioning-emotional, and mental health scale differed between countries, but these differences were not consistent. Wheelchair use was associated with lower physical and social functioning scores (B = -23.6 and 15.1, p < 0.001), and the use of artificial ventilation with lower physical functioning scores (B = -8.4, p = 0.004). Patients reported significantly poorer quality of life than the general population on the physical functioning, role functioning-physical, general health, vitality, and social functioning scales. No significant differences in SF-36 scores were found between the baseline and 1 year follow-up measurement. CONCLUSIONS: Patients with late-onset Pompe disease are, on average, markedly affected on the physical health domains of quality of life, but score only slightly lower than the general population on the mental health domains. PMID- 15534257 TI - Important observations prior to the description of the Hoover sign. AB - OBJECTIVE: To study the context in which Hoover described his sign for differentiating hysterical and organic hemiplegia. BACKGROUND: At the turn of the 20th century, many physicians were looking for signs to distinguish organic from hysterical hemiparesis. In 1908, Hoover described his sign of "complementary opposition." Other signs based on associated movements of the upper extremities, lower extremities, and trunk were also described during and before this period and might have contributed to Hoover's understanding of complementary opposition. METHODS: A complete literature review of the original relevant articles by Babinski, Bychowski, Grasset and Gaussel, and Hoover was performed. RESULTS: Several similar maneuvers were described before the Hoover sign. Babinski described the trunk-thigh test in 1897. Bychowski in Warsaw (performed since 1902, published in 1907) and Grasset and Gaussel in Paris (1905) independently described a phenomenon in which the separate elevation of each leg was performed more easily than simultaneous elevation. Moreover, Bychowski, although not emphasizing it as a sign, described what became known as Hoover sign. Hoover sign (1908) has been considered a further elaboration of these previous observations. Interestingly, Hoover had studied in Europe, and although no direct relationship between his discovery and these observations has been elucidated, the authors postulate that he was influenced by Babinski, Bychowski, and Grasset and Gaussel. CONCLUSION: Although Babinski, Bychowski, and Grasset and Gaussel described similar signs, only Hoover sign is still in wide use by practitioners. PMID- 15534258 TI - T2*-weighted MRI in cortical venous thrombosis. PMID- 15534259 TI - Different progression rates for deep white matter hyperintensities in elderly men and women. AB - The authors investigated the progression of white matter hyperintensities (WMHs) in a large population of elderly men and women. After 3 years of follow-up, women had accumulated approximately twice as much deep WMH (DWMH) as men. The progression of periventricular WMH was the same for men and women. Gender differences may affect the pathogenesis of DWMH, which in turn may result in different clinical consequences in women. PMID- 15534260 TI - A presenilin 1 R278I mutation presenting with language impairment. AB - Presenilin (PSEN)1 mutations are responsible for many cases of autosomal dominant Alzheimer disease (AD), although the clinical spectrum has not been fully defined. The authors describe two members of a kindred with a novel PSEN1 mutation (R278I) presenting with language impairment and relative preservation of memory. Screening for PSEN1 mutations may be appropriate in cases of familial dementia even where the clinical phenotype is not typical of AD. PMID- 15534261 TI - Impact of antioxidants, zinc, and copper on cognition in the elderly: a randomized, controlled trial. AB - Participants in the Age-Related Eye Disease Study were randomly assigned to receive daily antioxidants (vitamin C, 500 mg; vitamin E, 400 IU; beta carotene, 15 mg), zinc and copper (zinc, 80 mg; cupric oxide, 2 mg), antioxidants plus zinc and copper, or placebo. A cognitive battery was administered to 2,166 elderly persons after a median of 6.9 years of treatment. Treatment groups did not differ on any of the six cognitive tests (p > 0.05 for all). These results do not support a beneficial or harmful effect of antioxidants or zinc and copper on cognition in older adults. PMID- 15534262 TI - Cervical artery dissection: An atypical presentation with Ehlers-Danlos-like collagen pathology? AB - The authors took skin biopsies of the macroscopically normal skin of seven consecutive patients with spontaneous cervical artery dissection (SCAD). Histologically, alterations of the collagen and elastic fiber networks were found in six patients. In five, the histologic, immunohistochemical, and ultrastructural changes were similar to those usually found in Ehlers-Danlos syndrome (EDS). This suggests that SCAD is frequently associated with the dermal alterations seen in EDS. PMID- 15534263 TI - Alcohol dehydrogenase 2 variant is associated with cerebral infarction and lacunae. AB - The authors examined the association of the alcohol dehydrogenase 2 (ADH2) genotype with vascular events in community-dwelling Japanese (1,102 men/1,093 women). The allele ADH2*2 encodes an isozyme with a higher level of activity than ADH2*1. Here, the authors show that the ADH2*1 carriage is associated with high prevalence of cerebral infarction and lacunae in men. Multiple regression analyses confirmed that the risk of lacunae and cerebral infarction was increased by the ADH2*1 allele. PMID- 15534264 TI - Treatment of trigeminal neuralgia associated with multiple sclerosis: case report. AB - The incidence of trigeminal neuralgia (TN) in patients with multiple sclerosis is higher than in the general population. If pharmacologic therapy fails, invasive procedures in the gasserian ganglion are usually offered. Microvascular decompression is not routinely performed. The authors report a patient with persistent TN after repetitive trigeminal radiofrequency rhizotomy and finally successful microvascular decompression after demonstration of neurovascular compression with high-resolution MRI. PMID- 15534265 TI - Relation between objective and subjective measures of bladder dysfunction in multiple sclerosis. AB - The authors studied 297 patients with multiple sclerosis (MS), correlating urinary symptoms (bowel/bladder Functional System [FS] score of the Expanded Disability Status Scale [EDSS] and bladder dysfunction score of the Guy's Neurological Disability Scale [GNDS[) vs objective measurement of bladder dysfunction (postmicturition residual volume). EDSS and GNDS were of no value for predicting the presence of a clinically relevant postvoiding volume. Therefore, the authors recommend ultrasound scanning of residual volume in every patient with MS, even in the absence of subjective urinary symptoms. PMID- 15534266 TI - Post-infectious encephalomyelitis associated with St. Louis encephalitis virus infection. AB - Neurologic illness associated with acute St. Louis encephalitis, West Nile, and Japanese encephalitis virus infection includes acute aseptic meningitis, encephalomyelitis, and a poliomyelitis-like syndrome. Few post-infectious immune mediated neurologic events associated with flaviviral infection have been reported. The authors report on a woman with apparent post-infectious encephalomyelitis associated with recent St. Louis encephalitis virus infection, suggesting that neurologic illness from flaviviruses may also be seen in the post infectious period following mild clinical illness. PMID- 15534267 TI - Hypertrophic pachymeningitis as an initial and cardinal manifestation of microscopic polyangiitis. AB - The authors describe a 68-year-old man who developed hypertrophic pachymeningitis as an initial and cardinal manifestation of microscopic polyangiitis. The patient had a high titer of antineutrophil cytoplasmic antibody for myeloperoxidase. Biopsies revealed necrotizing glomerulonephritis in the kidney, small vessel vasculitis in the sural nerve, and infiltration of plasma cells and eosinophils in the thickened dura mater. PMID- 15534268 TI - Paroxysmal alien limb phenomena due to epileptic seizures and electrical cortical stimulation. AB - Alien limb phenomena (ALPs) are characterized by limb movements, which are subjectively experienced as involuntary or alien induced. ALPs regularly remain unchanged and occur as a consequence of frontal, callosal, or posterior cerebral lesions. The authors present two patients with paroxysmal ALP proved to be focal seizures by using video-EEG monitoring. In another two patients, ALP could experimentally be induced by electrical cortical stimulation. Based on the stimulation results, the authors suspect a functional disconnection of 1) sensory cortical areas providing information about the extrapersonal space; and 2) areas of the frontal and/or limbic cortex that are regularly involved in the processing and executing of intentional motor activity as a pathophysiologic substrate for ictal ALP. PMID- 15534269 TI - Short-term outpatient EEG video with induction in the diagnosis of psychogenic seizures. AB - To analyze the yield of short-term outpatient EEG video monitoring, the authors reviewed data on all patients who underwent this procedure at their center. All patients were suspected of having psychogenic nonepileptic seizures (PNES) on clinical grounds. The total number of cases of short-term outpatient EEG video monitoring was 74. In 49 (66%) cases, the suspected diagnosis of PNES could be confirmed, thereby obviating the need for prolonged inpatient EEG video monitoring. PMID- 15534270 TI - Diffuse neurodeficits in intravascular lymphomatosis with ADAMTS13 inhibitor. AB - Inhibitory antibody to von Willebrand factor (vWF)-cleaving protease (ADAMTS13) was detected in a patient with intravascular lymphomatosis. The increased serum level of the antibody paralleled an increase in the expression of uncleaved vWF, which might cause microvascular thrombosis and platelet consumption. Malignant cell proliferations with superimposed thrombosis within the lumina throughout the entire vasculature account for diffuse neurodeficits observed in the patient. PMID- 15534271 TI - Autoimmune etiology of complex regional pain syndrome (M. Sudeck). AB - Sera of 12 patients with complex regional pain syndrome (CRPS) were tested for the occurrence of autoantibodies against nervous system structures. Immunohistochemistry revealed autoantibodies against autonomic nervous system structures in 5 of 12 (41.6%) of the patients. Western blot analysis showed neuronal reactivity in 11 of 12 (91.6%) patients. The authors hypothesize that CRPS can result from an autoimmune process against the sympathetic nervous system. PMID- 15534272 TI - Idiopathic intracranial hypertension: risk of recurrences. AB - Long-term prognosis and visual outcome of 54 patients with idiopathic intracranial hypertension (IIH) was studied. Mean observation period was 6.2 years; 33 patients had two or more recurrences. Visual acuity was preserved in all patients without recurrence and in 86% of patients with recurrences. Recurrences did not occur while patients were maintained on acetazolamide. No statistical difference was found between IIH patients who had only one event, compared to the recurrent group. PMID- 15534273 TI - The autonomic and behavioral profile of emotional dysregulation. AB - The authors describe a patient with focal brain atrophy and emotional lability characterized by episodes of excessive crying and laughing. The patient was selectively impaired in the production of voluntary complex facial movements and was unable to regulate her emotional behavior and autonomic reactivity. She also displayed increased behavioral and autonomic changes when explicitly trying to suppress her responses to emotional stimuli (compared with when not trying to regulate her responses). This pattern of deficits supports a selective deficit in voluntary emotional control. PMID- 15534274 TI - Adult alpha-mannosidosis: clinical progression in the absence of demyelination. AB - Alpha-mannosidosis is an inherited lysosomal storage disease. The authors report three siblings (ages 38 to 47 years) with the rare adult variant. All three had late-onset ataxia and retinal degeneration, adding to hearing loss, cognitive impairment, and dysotosis multiplex. One sibling also had psychosis. MRI revealed cerebellar atrophy and predominantly parieto-occipital white matter changes. MR spectroscopy showed no evidence for demyelination. It appears that the disabling course of adult alpha-mannosidosis is caused by lysosomal accumulation rather than demyelination. PMID- 15534275 TI - The occurrence of Guillain-Barre syndrome within families. AB - This report describes 12 Dutch families of which at least two members have had Guillain-Barre syndrome (GBS). The authors observed an earlier onset of GBS in successive generations. The occurrence of GBS within families suggests a role for genetic factors in the pathogenesis of GBS. PMID- 15534276 TI - Sciatic neurolymphomatosis. PMID- 15534277 TI - Acquired generalized anhidrosis associated with lymphocytic infundibuloneurohypophysitis. PMID- 15534278 TI - Unilateral localized hyperhidrosis associated with frontal lobe meningioma. PMID- 15534279 TI - Acute neuropathy with multiple conduction blocks after TNFalpha monoclonal antibody therapy. PMID- 15534280 TI - Intervertebral disk and vertebral body infarction as a confirmatory sign of spinal cord ischemia. PMID- 15534281 TI - Early treatment of Aicardi syndrome with vigabatrin can improve outcome. PMID- 15534282 TI - Tinnitus evoked by cutaneous stimulation. PMID- 15534283 TI - Assessing the neurology clerkship: the value of student feedback. PMID- 15534285 TI - Percutaneous closure of patent foramen ovale reduces the frequency of migraine attacks. PMID- 15534284 TI - Brainstem involvement in hypertensive encephalopathy: clinical and radiological findings. PMID- 15534286 TI - Paraparesis due to capsular hemorrhages. PMID- 15534287 TI - Experience with immunomodulatory treatments in Rasmussen's encephalitis. PMID- 15534288 TI - Antiplatelet drug discontinuation is a risk factor for ischemic stroke. PMID- 15534289 TI - Global failures. PMID- 15534290 TI - Incomplete patient drug information still a problem. PMID- 15534292 TI - Catering to the customers. PMID- 15534295 TI - Care at for-profit hospitals. PMID- 15534296 TI - Care at for-profit hospitals. PMID- 15534298 TI - The gender gap in Canadian health care. PMID- 15534299 TI - Reforms mean 25 million Russians lose free health care. PMID- 15534300 TI - Mandatory error reporting in Saskatchewan. PMID- 15534301 TI - Isolate patients, screen staff to fight MRSA. PMID- 15534302 TI - Obesity epidemic migrates east. PMID- 15534303 TI - SARS silver lining: a renewal of public health. PMID- 15534304 TI - Obstacles to combatting cancer in the developing world. PMID- 15534305 TI - Varicella vaccine cost-effective: US study. PMID- 15534306 TI - Learning to care for those who can't be cured. PMID- 15534307 TI - Vagus nerve stimulation in the treatment of epilepsy. PMID- 15534309 TI - A 32-year-old woman with breast swelling and crepitant rales. PMID- 15534308 TI - Prevention of influenza in the general population: recommendation statement from the Canadian Task Force on Preventive Health Care. PMID- 15534310 TI - Optimal search strategies for detecting health services research studies in MEDLINE. AB - BACKGROUND: Evidence from health services research (HSR) is currently thinly spread through many journals, making it difficult for health services researchers, managers and policy-makers to find research on clinical practice guidelines and the appropriateness, process, outcomes, cost and economics of health care services. We undertook to develop and test search terms to retrieve from the MEDLINE database HSR articles meeting minimum quality standards. METHODS: The retrieval performance of 7445 methodologic search terms and phrases in MEDLINE (the test) were compared with a hand search of the literature (the gold standard) for each issue of 68 journal titles for the year 2000 (a total of 25,936 articles). We determined sensitivity, specificity and precision (the positive predictive value) of the MEDLINE search strategies. RESULTS: A majority of the articles that were classified as outcome assessment, but fewer than half of those in the other categories, were considered methodologically acceptable (no methodologic criteria were applied for cost studies). Combining individual search terms to maximize sensitivity, while keeping specificity at 50% or more, led to sensitivities in the range of 88.1% to 100% for several categories (specificities ranged from 52.9% to 97.4%). When terms were combined to maximize specificity while keeping sensitivity at 50% or more, specificities of 88.8% to 99.8% were achieved. When terms were combined to maximize sensitivity and specificity while minimizing the differences between the 2 measurements, most strategies for HSR categories achieved sensitivity and specificity of at least 80%. INTERPRETATION: Sensitive and specific search strategies were validated for retrieval of HSR literature from MEDLINE. These strategies have been made available for public use by the US National Library of Medicine at www.nlm.nih.gov/nichsr/hedges/search.html. PMID- 15534311 TI - Evidence of suboptimal management of cardiovascular risk in patients with type 2 diabetes mellitus and symptomatic atherosclerosis. AB - BACKGROUND: Given that most deaths among patients with diabetes mellitus are due to cardiovascular disease, we sought to determine the extent to which medications proven to reduce cardiovascular mortality are prescribed for patients with type 2 diabetes who have symptomatic atherosclerosis (i.e., coronary artery disease [CAD], cerebrovascular disease [CBVD] or peripheral arterial disease [PAD]). METHODS: Administrative records from Saskatchewan Health were used to evaluate the use of antiplatelet agents, statins and angiotensin-converting enzyme (ACE) inhibitors by people with treated type 2 diabetes with and without symptomatic atherosclerosis. CAD and CBVD were defined by International Classification of Diseases (ninth revision) codes, and PAD was defined on the basis of pentoxifylline use or lower limb amputation. Multivariate logistic regression analysis was used to compare medication use in patients with and without PAD, with adjustments for differences in age, sex and comorbidity. RESULTS: In this cohort of 12,106 patients with type 2 diabetes (mean age 64 years, 55% male, mean follow-up 5 years), fewer than 25% received an antiplatelet agent or statin, and fewer than 50% received an ACE inhibitor. Although patients with CAD were more likely to receive antiplatelet agents, statins or ACE inhibitors than people without CAD (p < 0.001 for all), the overall use of these medications was suboptimal (37%, 29% and 60% respectively among patients with symptomatic CAD). Similar patterns of practice were found for patients with symptomatic CBVD and PAD. All 3 proven efficacious therapies were prescribed for only 11% of patients with CAD, 22% with CBVD and 12% with PAD. Patients with PAD who had undergone lower limb amputation were no more likely to subsequently receive antiplatelet agents or statins than those without an amputation. INTERPRETATION: Diabetic patients with symptomatic atherosclerotic disease are undertreated with medications known to reduce cardiovascular morbidity and mortality, perhaps because of a "glucocentric" view of diabetes. Programs to improve the quality of cardiovascular risk reduction in these high-risk patients are needed. PMID- 15534312 TI - Predictive value of a self-reported history of varicella infection in determining immunity in adults. AB - The recent introduction of a vaccine for varicella has raised questions about whether, for adults, a patient's history of varicella infection is useful in determining if vaccination is necessary. We report findings on 184 family medicine patients aged 18 to 65 years who were asked if they had a history of varicella infection and were subsequently tested for varicella antibodies. A history of infection was positive for 114 (62%) of the participants and negative or uncertain for 70 (38%). All 114 subjects who reported a varicella infection history were immune. All 4 subjects who were not immune reported an uncertain or negative infection history. Except for people who are at increased risk of varicella infection or complications from infection, serologic testing may not be required for adults in the general population who have a history of varicella infection. PMID- 15534313 TI - Global health indicators: an overview. PMID- 15534314 TI - Health and economic benefits of an accelerated program of research to combat global infectious diseases. PMID- 15534315 TI - Prevention of influenza in the general population. AB - BACKGROUND: Although all jurisdictions in Canada offer annual influenza immunization to people at high risk of complications, only Ontario has provided universal annual immunization of healthy adults and children. Use of chemotherapy (amantidine, neuraminidase inhibitors) to prevent influenza varies among provinces. We sought to systematically review the evidence for the prevention of influenza infection in the general population. METHODS: The interventions reviewed were influenza vaccination and prophylactic use of neuraminidase inhibitors. The health outcomes of interest were rates of laboratory-confirmed influenza infection, clinical definitions of influenza-like illness and work absenteeism. MEDLINE and Cochrane databases were searched for relevant articles published between 1966 and March 2003. Only randomized controlled trials (RCTs) were selected. Evidence was appraised using the methodology of the Canadian Task Force on Preventive Health Care. RESULTS: Eighteen trials involving more than 33,000 healthy adults were identified that met the inclusion criteria; of these, 15 showed that influenza vaccination with either live-attenuated and inactivated vaccines was efficacious. Eleven trials were considered to be of "good" quality, and 7 were considered to be of "fair" quality. The relative risk reduction (RRR) associated with influenza immunization in adults ranged from 0% to 91%. Fifteen RCTs involving more than 45,000 healthy children aged 6 months to 19 years were identified, of which 9 were considered to contain "good" evidence and 6 "fair" evidence. Results from 12 of these trials showed protection against influenza. The RRR ranged from 0% to 93%. There were 6 RCTs of "good" quality showing that neuraminidase inhibitors are effective in preventing influenza infection. Side effects from both influenza vaccination and neuraminidase inhibitor administration were mild. INTERPRETATION: There are numerous RCTs of good quality in large populations that have consistently shown that influenza vaccination, using inactivated or live-attenuated vaccines, is moderately effective in preventing influenza in the general population (healthy adults and children over 6 months of age). There is good evidence that neuraminidase inhibitor prophylaxis in contacts given within 36 to 48 hours of symptom onset of the household index case is effective; appropriate use of this prevention method requires access to rapid diagnostic methods. Decisions about introduction of routine immunization programs must take into account the cost and cost-effectiveness of a universal program and the burden of illness associated with influenza in each jurisdiction. PMID- 15534316 TI - Transcription regulation within the circadian clock: the E-box and beyond. AB - The circadian oscillator is composed of transcriptional feedback loops in organisms ranging from cyanobacteria to humans. These transcriptional feedback loops are so named because transcriptional regulators accumulate to high levels and then feed back to control their own genes' transcription, thus generating a self-sustaining rhythm in gene expression. In insects and vertebrates, the genes that encode these feedback regulators are remarkably well conserved and function to control not 1 but 2 feedback loops. These feedback loops control rhythmic transcription in opposite phases of the circadian cycle, yet they are interlocked because they share a number of components. In this review, the author will compare transcriptional regulatory mechanisms within the Drosophila and mammalian feedback loops and outline remaining questions concerning transcriptional regulation within and downstream of these feedback loops. PMID- 15534317 TI - Posttranscriptional and posttranslational regulation of clock genes. AB - Circadian rhythms have been observed in diverse organisms, including plants, animals, bacteria, and fungi. In such organisms, the circadian clock is primarily composed of a cell-autonomous transcriptional feedback loop. In addition to transcriptional regulation, the modification of core clock transcripts and proteins can dramatically affect the circadian clock. In this review, the authors discuss some of the posttranscriptional and posttranslational modifications and their effects on the circadian clock. The combined outcome of these modifications is to adjust the timing of the clock to produce a circadian oscillator that takes approximately 24 h. PMID- 15534318 TI - It's all in the timing: many clocks, many outputs. AB - It is thought that circadian regulation of physiology and behavior imparts survival advantages to organisms that use clocks. In mammals, a master clock resident in the SCN synchronizes other central and peripheral oscillators to evoke this regulation. This master oscillator consists of interlocking transcriptional-translational feedback loops, and it regulates both core clock genes necessary for oscillator maintenance as well as specific output genes that directly or indirectly mediate physiology under circadian control. It is now clear that both neuroanatomic and molecular outputs of the clock are necessary for proper circadian clock function. Recent technology has improved our understanding of these processes, elucidating the anatomic outputs of the SCN, as well as the molecular outputs of both central and peripheral oscillators that mediate observed physiological changes. PMID- 15534319 TI - Clock genes in cell clocks: roles, actions, and mysteries. AB - Cellular events must be organized in the time dimension as well as in the space dimension for many proteins to perform their cellular functions effectively. The intracellular molecular oscillating loops that compose the cell's circadian clock coordinate the timing of the expression of a variety of genes with basic or specific cellular functions. In mammals, the temporal pattern of clock gene expression generated in each SCN neuron is coupled to those of other cells and, amplified, spreads its signals through the brain and then, via feeding behavior, glucocorticoids, and sympathetic nerves, to peripheral organs. These peripheral organs have their own circadian clocks. In some tissues, such as liver, there is also a clock-regulating cell cycle, which interacts strongly with the components and temporal organization of the circadian clock. Some tissues, however, such as testis, express clock genes whose function, if any, remains unclear. Furthermore, circadian clock function may be suspended in differentiating tissue. Thus, the prominence of circadian organization may not apply equally to all tissues under all conditions. PMID- 15534320 TI - Clock genes, oscillators, and cellular networks in the suprachiasmatic nuclei. AB - The mammalian SCN contains a biological clock that drives remarkably precise circadian rhythms in vivo and in vitro. Recent advances have revealed molecular and cellular mechanisms required for the generation of these daily rhythms and their synchronization between SCN neurons and to the environmental light cycle. This review of the evidence for a cell-autonomous circadian pacemaker within specialized neurons of the SCN focuses on 6 genes implicated within the pace making mechanism, an additional 4 genes implicated in pathways from the pacemaker, and the intercellular and intracellular mechanisms that synchronize SCN neurons to each other and to solar time. PMID- 15534321 TI - The neurospora circadian system. AB - The eukaryotic filamentous fungus Neurospora crassa has proven to be a durable and dependable model system for the analysis of the cellular and molecular bases of circadian rhythms. Pioneering genetic analyses identified clock genes, and beginning with the cloning of frequency (frq), work over the past 2 decades has revealed the molecular basis of a core circadian clock feedback loop that has illuminated our understanding of circadian oscillators in microbes, plants, and animals. In this transcription/translation-based feedback loop, a heterodimer of the White Collar-1 (WC-1) and WC-2 proteins acts both as the circadian photoreceptor and, in the dark, as a transcription factor that promotes the expression of the frq gene. FRQ dimerizes and feeds back to block the activity of its activators (making a negative feedback loop), as well as feeding forward to promote the synthesis of its activator, WC-1. Phosphorylation of FRQ by several kinases leads to its ubiquitination and turnover, releasing the WC-1/WC-2 dimer to reactivate frq expression and restart the circadian cycle. Light resetting of the clock can be understood through the rapid light induction of frq expression and temperature resetting through the influence of elevated temperatures in driving higher levels of FRQ. Several FRQ- and WC-independent, noncircadian FRQ less oscillators (FLOs) have been described, each of which appears to regulate aspects of Neurospora growth or development. Overall, the FRQ/white collar complex feedback loop appears to coordinate the circadian system through its activity to regulate downstream-target clock-controlled genes, either directly or via regulation of driven FLOs. PMID- 15534322 TI - The Arabidopsis thaliana clock. AB - A combination of forward and reverse genetic approaches together with transcriptome-scale gene expression analyses have allowed the elaboration of a model for the Arabidopsis thaliana circadian clock. The working model largely conforms to the expected negative feedback loop model that has emerged from studies in other model systems. Although a core loop has emerged, it is clear that additional components remain to be identified and that the workings of the Arabidopsis clock have been established only in outline. Similarly, the details of resetting by light and temperature are only incompletely known. In contrast, the mechanism of photoperiodic induction of flowering is known in considerable detail and is consistent with the external coincidence model. PMID- 15534323 TI - Circadian timing mechanism in the prokaryotic clock system of cyanobacteria. AB - Cyanobacteria are the simplest organisms known to exhibit circadian rhythms and have provided experimental model systems for the dissection of basic properties of circadian organization at the molecular, physiological, and ecological levels. This review focuses on the molecular and genetic mechanisms of circadian rhythm generation in cyanobacteria. Recent analyses have revealed the existence of multiple feedback processes in the prokaryotic circadian system and have led to a novel molecular oscillator model. Here, the authors summarize current understanding of, and open questions about, the cyanobacterial oscillator. PMID- 15534324 TI - Clock gene evolution and functional divergence. AB - In considering the impact of the earth's changing geophysical conditions during the history of life, it is surprising to learn that the earth's rotational period may have been as short as 4 h, as recently as 1900 million years ago (or 1.9 billion years ago). The implications of such figures for the origin and evolution of clocks are considerable, and the authors speculate on how this short rotational period might have influenced the development of the "protoclock" in early microorganisms, such as the Cyanobacteria, during the geological periodsin which they arose and flourished. They then discuss the subsequent duplication of clock genes that took place around and after the Cambrian period, 543 million years ago, and its consequences. They compare the relative divergences of the canonical clock genes, which reveal the Per family to be the most rapidly evolving. In addition, the authors use a statistical test to predict which residues within the PER and CRY families may have undergone functional specialization. PMID- 15534325 TI - Toward a national research agenda on violence against women: continuing the dialogue on research and practice. PMID- 15534326 TI - What is violence against women: defining and measuring the problem. AB - Violence against women (VAW) is a prevalent problem with substantial physical and mental health consequences throughout the world, and sound public policy is dependent on having good measures of VAW. This article (a) describes and contrasts criminal justice and public health approaches toward defining VAW, (b) identifies major controversies concerning measurement of VAW, (c) summarizes basic principles in identifying and measuring VAW cases, and (d) recommends changes to improve measurement of VAW. In addition to reviewing recommendations from the Centers for Disease Control and Prevention Workshop on Building Data Systems for Monitoring and Responding to Violence Against Women and the World Health Organization World Report on Violence and Health, the article concludes that changes are needed in the FBI Uniform Crime Reports and National Crime Victimization Survey to improve measurement of rape and sexual assault. PMID- 15534327 TI - Issues related to defining and measuring violence against women: response to Kilpatrick. AB - This paper asserts that although there is considerable agreement in the U.S. and internationally about the importance of uniform terminology and measurement related to violence against women, we need a strategy for choosing standardized definitions and measures. Responding to Kilpatrick's comments at the October 2003 national research conference on violence against women, the author stresses the importance of developing and using uniform terminology related to violence against women, and discusses the lack of a formal mechanism to achieve uniformity of definitions and measurement. Uncertainty about the impact of context on survey findings and the lack of agreement about the optimal scope of measurement are discussed. The author also comments on some difficulties associated with implementing Kilpatrick's proposed modifications to existing measures of rape and sexual assault. PMID- 15534328 TI - What is violence against women? Defining and measuring the problem: a response to Dean Kilpatrick. AB - This article discusses (a) the relationship between the reemergence of the women's movement in the 1970s and the current field of violence against women; (b) the role voluntary associations and social activism play in shaping social institutions, including the institution of academic scholarship; (c) the need to broaden our definition of violence against women to include nonviolent acts, such as stalking and emotional abuse; (d) the need to study the interrelationships among various forms of violence and abuse, including violence and abuse experienced as a minor and violence and abuse experienced as an adult; (e) the importance of using precise language when reporting research findings; (f) the importance of using a multiple definition/measurement approach when conducting research on violence against women; and (g) the need for comprehensive research on violence against women that takes into account the various types of violent and abusive behaviors experienced by women over their lifetimes. PMID- 15534329 TI - Violence against women: outcome complexity and implications for assessment and treatment. AB - This article reviews the major forms of violence against women, including sexual assault, intimate-relationship violence, and stalking and outlines the known psychological effects of such victimization. Also discussed are a number of variables that combine to determine the effects of such victimization, including type and characteristics of the assault; victim variables such as demographics, psychological reactions at the time of the trauma, previous victimization history, current or previous psychological difficulties, and general coping style; and sociocultural factors such as poverty, social inequality, and inadequate social support. The implications of this complexity are explored in terms of psychological assessment and the frequent need for multitarget, multimodal treatment approaches. PMID- 15534330 TI - Complexity of women's response to violence: response to Briere and Jordan. PMID- 15534331 TI - Beyond PTSD: mental health consequences of violence against women: a response to Briere and Jordan. AB - This article proposes that we move beyond posttraumatic stress disorder (PTSD) in our conceptualization of traumatic stress responses of victimized women exposed to serial forms of unrelenting violence, such as intimate partner violence and stalking. It is argued that the traditional PTSD framework is ill fitting in the context of some forms of violence against women (VAW), and these limits have consequences for developing appropriate interventions for some victimized women. The article further suggests going beyond PTSD by developing a more nuanced understanding of the ways in which PTSD and other mental health symptoms contribute to the vast array of deleterious personal, societal, and economic costs of VAW. PMID- 15534332 TI - A suggested research agenda on treatment-outcome research for female victims of violence. PMID- 15534333 TI - Intimate partner violence and physical health consequences: policy and practice implications. AB - Extensive research indicates that intimate partner violence (IPV) poses a significant risk to the physical health of women. IPV is associated with increased mortality, injury and disability, worse general health, chronic pain, substance abuse, reproductive disorders, and poorer pregnancy outcomes. IPV is also associated with an overuse of health services and unmet need for services, as well as strained relationships with providers. The body of IPV research has several critical gaps. There are almost no longitudinal studies of IPV and health. Most studies are clustered into a few specialties, with almost no research in the areas of allied health, dentistry, or management. A common definition of IPV is still not used. Finally, with some notable exceptions, there has been little success in moving the health care system to routinely screen women for IPV. PMID- 15534334 TI - Primary prevention of intimate partner violence for women's health: a response to Plichta. PMID- 15534335 TI - Intimate partner violence and physical health consequences: commentary on Plichta. PMID- 15534336 TI - Toward developing complex multivariate models for examining the intimate partner violence-physical health relationship: commentary on Plichta. PMID- 15534337 TI - Researcher-advocate collaborations to end violence against women: toward liberating methodologies for action research. AB - This article relies on literature from the field and on research conducted by the National Violence Against Women Prevention Research Center to describe the difficulties encountered in collaborations between researchers and advocates working on the prevention of violence against women. The article summarizes guidelines for collaboration that have been suggested by many researchers and advocates, and it makes the claim that a feminist approach to liberating methodologies for action research is needed to increase the likelihood of successful collaborations. PMID- 15534338 TI - Reader variability in mammography and its implications for expected utility over the population of readers and cases. AB - The multiple-reader, multiple-case (MRMC) approach to receiver operating characteristic (ROC) analysis is becoming the dominant assessment paradigm in medical imaging. Its most common version involves having many readers read every patient case in the study, a critical feature since differences among competing imaging modalities are often dominated by differences in reader performance. The present authors have carried out MRMC ROC analysis on a uniquely large data set for mammography. The analysis quantifies the great range of observed reader skill in that data set. It also demonstrates that the sample sizes are sufficiently large that the conclusions generalize to the populations sampled here with little uncertainty from the finite sample size. A schematic approach to bracketing the utility matrix is then used to study trends in the resulting expected utility functions that correspond to the range of observed ROC curves. This is done for both the screening and the diagnostic context. The results raise 2 hypotheses for further investigation. First, it is possible that the present ambiguity surrounding the effectiveness of mammography is due in part to the observed range of reader skills and corresponding expected utility functions. Second, it is possible that computer-assisted modalities for mammography may lead to improvements in the expected utility function not only for screening but also in the diagnostic context, especially for the lower performing readers. PMID- 15534339 TI - The application of the heuristic-systematic processing model to treatment decision making about prostate cancer. AB - The study investigated the utility of the Heuristic-Systematic Processing Model as a framework for the investigation of patient decision making. A total of 111 men recently diagnosed with localized prostate cancer were assessed using Verbal Protocol Analysis and self-report measures. Study variables included men's use of nonsystematic and systematic information processing, desire for involvement in decision making, and the individual differences of health locus of control, tolerance of ambiguity, and decision-related uncertainty. Most men (68%) preferred that decision making be shared equally between them and their doctor. Men's use of the expert opinion heuristic was related to men's verbal reports of decisional uncertainty and having a positive orientation to their doctor and medical care; a desire for greater involvement in decision making was predicted by a high internal locus of health control. Trends were observed for systematic information processing to increase when the heuristic strategy used was negatively affect laden and when men were uncertain about the probabilities for cure and side effects. There was a trend for decreased systematic processing when the expert opinion heuristic was used. Findings were consistent with the Heuristic-Systematic Processing Model and suggest that this model has utility for future research in applied decision making about health. PMID- 15534340 TI - A cost-utility analysis of neonatal circumcision. AB - A cost-utility analysis, based on published data from multiple observational studies, comparing boys circumcised at birth and those not circumcised was undertaken using the Quality of Well-being Scale, a Markov analysis, the standard reference case, and a societal perspective. Neonatal circumcision increased incremental costs by $828.42 per patient and resulted in an incremental 15.30 well-years lost per 1000 males. If neonatal circumcision was cost-free, pain free, and had no immediate complications, it was still more costly than not circumcising. Using sensitivity analysis, it was impossible to arrange a scenario that made neonatal circumcision cost-effective. Neonatal circumcision is not good health policy, and support for it as a medical procedure cannot be justified financially or medically. PMID- 15534341 TI - Decision making in a multidisciplinary cancer team: does team discussion result in better quality decisions? AB - To establish whether treatment recommendations made by clinicians concur with the best outcomes predicted from their prognostic estimates and whether team discussion improves the quality or outcome of their decision making, the authors studied real-time decision making by a lung cancer team. Clinicians completed pre and postdiscussion questionnaires for 50 newly diagnosed patients. For each patient/doctor pairing, a decision model determined the expected patient outcomes from the clinician's prognostic estimates. The difference between the expected utility of the recommended treatment and the maximum utility derived from the clinician's predictions of the outcomes (the net utility loss) following all potential treatment modalities was calculated as an indicator of quality of the decision. The proportion of treatment decisions changed by the multidisciplinary team discussion was also calculated. Insofar as the change in net utility loss brought about by multidisciplinary team discussion was not significantly different from zero, team discussion did not improve the quality of decision making overall. However, given the modest power of the study, these findings must be interpreted with caution. In only 23 of 87 instances (26%) in which an individual specialist's initial treatment preference differed from the final group judgment did the specialist finally concur with the group treatment choice after discussion. This study does not support the theory that team discussion improves decision making by closing a knowledge gap. PMID- 15534342 TI - An exploration of relative health stock in advanced cancer patients. AB - OBJECTIVE: The authors sought to empirically test whether relative health stock, a measure of patients' sense of loss in their health due to illness, influences the treatment decisions of patients facing life-threatening conditions. Specifically, they estimated the effect of relative health stock on advanced cancer patients' decisions to participate in phase I clinical trials. METHOD: A multicenter study was conducted to survey 328 advanced cancer patients who were offered the opportunity to participate in phase I trials. The authors asked patients to estimate the probabilities of therapeutic benefits and toxicity, their relative health stock, risk preference, and the importance of quality of life. RESULTS: Controlling for health-related quality of life, an increase in relative health stock by 10 percentage points reduced the odds of choosing to participate in a phase I trial by 16% (odds ratio = 0.84, 95% confidence interval = 0.72, 0.97). CONCLUSION: Relative health stock affects advanced cancer patients' treatment decisions. PMID- 15534343 TI - A comparison of 3 valuation methods for temporary health states in patients treated with oral anticoagulants. AB - BACKGROUND: The application of the time tradeoff (TTO) method in temporary health states may lead to less valid results because an unrealistic scenario is presented to patients. The chained TTO has been proposed to solve this problem. OBJECTIVES: To compare a chained TTO method with a conventional TTO method in the valuation of temporary health states, in terms of consistency and reliability. To compare both TTO methods with direct rating. PATIENTS AND METHODS: Eighty-four patients treated with oral anticoagulants were interviewed twice. During the 1st interview, values for 5 temporary health states were obtained with a rank ordering procedure, direct rating, and the chained TTO method. During the 2nd interview, either the 1st interview was repeated (n = 30) or health state values were obtained with the conventional TTO method (n = 54). Consistency was assessed by comparing the 3 valuation methods with the rank ordering procedure. Generalizability theory was used to assess reliability. RESULTS: The 3 methods produced significantly different valuations of health states. Chained TTO values were higher than values obtained with direct rating and the conventional TTO. Consistency and reliability did not differ across the 3 methods. CONCLUSION: The authors found no evidence for a difference in consistency and reliability between the chained TTO method and the conventional TTO method in the valuation of temporary health states. As direct rating is simpler to administer than both TTO methods, one could consider using direct ratings for the valuation of temporary health states. Biases associated with the conventional and the chained TTO method are discussed. PMID- 15534344 TI - A Bayesian approach to net health benefits: an illustration and application to modeling HIV prevention. AB - PURPOSE: To conduct a cost-effectiveness analysis of HIV prevention when costs and effects cannot be measured directly. To quantify the total estimation of uncertainty due to sampling variability as well as inexact knowledge of HIV transmission parameters. METHODS: The authors focus on estimating the incremental net health benefit (INHB) in a randomized trial of HIV prevention with intervention and control conditions. Using a Bernoulli model of HIV transmission, changes in the participants' risk behaviors are converted into the number of HIV infections averted. A sampling model is used to account for variation in the behavior measurements. Bayes's theorem and Monte Carlo methods are used to attain the stated objectives. RESULTS: The authors obtained a positive mean INHB of 0.0008, indicating that advocacy training is just slightly favored over the control condition for men, assuming a $50,000 per quality-adjusted life year (QALY) threshold. To be confident of a positive INHB, the decision maker would need to spend more than $100,000 per QALY. PMID- 15534345 TI - On uncertainty in medical testing. AB - There is confusion in the medical decision-making literature about how to handle uncertainty in medical tests. In this article, the authors consider the situation in which there is uncertainty about the pretest probability of a disease in a patient as well as uncertainty about the sensitivity and specificity of a diagnostic test for that disease. They discuss how to calculate posttest probabilities of a disease under such uncertainty and how to calculate a distribution for a posttest probability. They show that given certain independence assumptions, uncertainty about these parameters need not complicate the calculation of patient positive predictive values: One can simply use the expected values of the parameters in the standard Bayesian formula for posttest probabilities. The discussion on how to calculate distributions for positive predictive values corrects a common and potentially important error. PMID- 15534346 TI - Combining correlated diagnostic tests: application to neuropathologic diagnosis of Alzheimer's disease. AB - This article studies the problem of combining correlated diagnostic tests to maximize the discriminating power between the diseased population and the healthy population. The authors consider all possible linear combinations of multiple diagnostic tests and search for the one that achieves the largest area under the receiver operating characteristic (ROC) curve. They discuss the statistical estimation of the optimum linear combination test and the associated maximum area under the ROC curve. Their approach is based on the assumption of multivariate normal distribution of the multiple diagnostic tests. They also present the application of the proposed techniques to the neuropathologic diagnosis of Alzheimer's disease based on brain lesions from 5 different brain locations using a data set from the Washington University Alzheimer's Disease Research Center. PMID- 15534347 TI - Alternative tree-structured survival analysis based on variance of survival time. AB - Tree-structured survival analysis (TSSA) is a popular alternative to the Cox proportional hazards regression in medical research of survival data. Several methods for constructing a tree of different survival profiles have been developed, including TSSA based on log-rank statistics, martingale residuals, Lp Wasserstein metrics between Kaplan-Meier survival curves, and a method based on a weighted average of the within-node impurity of the death indicator and the within-node loss function of follow-up times. Lu and others used variance of restricted mean lifetimes as an index of degree of separation (DOS) to measure the efficiency in separations of survival profiles by a classification method. Like tree-based regression analysis that uses variance as a criterion for node partition and pruning, the variance of restricted mean lifetimes between different groups can be an alternative index to log-rank test statistics in construction of survival trees. In this article, the authors explore the use of DOS in TSSA. They propose an algorithm similar to the least square regression tree for survival analysis based on the variance of the restricted mean lifetimes. They apply the proposed method to prospective cohort data from the Study of Osteoporotic Fracture that motivated the research and then compare their classification rule to those rules based on the conventional TSSA mentioned above. A limited simulation study suggests that the proposed algorithm is a competitive alternative to the log-rank or martingale residual-based TSSA approaches. PMID- 15534348 TI - Theory about heuristic strategies based on verbal protocol analysis: the emperor needs a shave. AB - A research report in the present issue of Medical Decision Making raises questions about the definition of heuristic strategies, the validity of conclusions drawn from the analysis of verbal protocols, and the tradeoff between rigor and relevance in research on the psychology of patient decision making. PMID- 15534350 TI - Neonatal circumcision: cost-effective preventive measure or "the unkindest cut of all"? PMID- 15534357 TI - Phase I trial of iodine-131 tositumomab with high-dose chemotherapy and autologous stem-cell transplantation for relapsed non-Hodgkin's lymphoma. AB - PURPOSE: To determine the maximum outpatient dose of iodine-131 tositumomab (up to 0.75 Gy) combined with high-dose carmustine, etoposide, cytarabine, and melphalan (BEAM) followed by autologous stem-cell transplantation (ASCT) for the treatment of chemotherapy-resistant relapsed or refractory B-cell non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Twenty-three patients with chemotherapy refractory or multiply-relapsed B-cell NHL were treated in a phase I trial combining iodine-131 tositumomab (ranging from 0.30 to 0.75 Gy total-body dose [TBD]) with high-dose BEAM followed by ASCT. RESULTS: The complete response rate after transplantation was 57%, and the overall response rate was 65%. Short-term and long-term toxicities were similar to historical control patients treated with BEAM alone. With a median follow-up of 38 months (range, 27 to 60 months), the overall survival (OS) rate was 55%, and the event-free survival (EFS) rate was 39%. CONCLUSION: There were no significant added toxicities apparent with the addition of iodine-131 tositumomab up to a dose of 0.75 Gy TBD to high-dose BEAM chemotherapy followed by ASCT. The EFS and OS were encouraging in this group of chemotherapy-resistant or refractory B-cell NHL patients. A follow-up phase II trial with iodine-131 tositumomab at the dose of 0.75 Gy TBD with BEAM is currently ongoing. PMID- 15534356 TI - Outcome of induction and postremission therapy in younger adults with acute myeloid leukemia with normal karyotype: a cancer and leukemia group B study. AB - PURPOSE: Evaluate the outcome of induction and postremission therapy in adults younger than 60 years with normal cytogenetics acute myeloid leukemia (AML). PATIENTS AND METHODS: In 490 patients, induction included cytarabine and daunorubicin (AD) or cytarabine and escalated doses of daunorubicin and etoposide +/- PSC-833 (ADE/ADEP). Intensification included one cycle of high-dose cytarabine (HDAC) followed by etoposide/cyclophosphamide and mitoxantrone/diaziquone (group I), three HDAC cycles (group II), four intermediate-dose cytarabine (IDAC) or HDAC cycles (group III), or one HDAC/etoposide cycle and autologous stem-cell transplantation (ASCT; group IV). RESULTS: Of 350 patients receiving AD, 73% achieved complete remission (CR), compared with 82% of 140 receiving ADE/ADEP (P = .04). Splenomegaly was associated with a lower CR rate (P < .001), and ADE/ADEP, with a higher CR rate in younger patients (P = .005). The 5-year disease-free survival (DFS) rate was 28% each for intensification groups I and II, compared with 41% and 45% for groups III and IV, respectively (P = .02). The 5-year cumulative incidence of relapse (CIR) was 62% and 67% for groups I and II, respectively, compared with 54% and 44% for groups III and IV, respectively (P = .049). The type of postremission intensification remained significant for DFS and CIR in multivariable analysis. CONCLUSION: In younger adults with normal cytogenetics AML, splenomegaly predicts a lower CR rate, and the postremission strategies of either four cycles of I/HDAC or one cycle of HDAC/etoposide followed by ASCT are associated with improved DFS and reduced relapse compared with therapies that include fewer cycles of cytarabine or no transplantation. PMID- 15534358 TI - Autologous and allogeneic stem-cell transplantation as salvage treatment of acute promyelocytic leukemia initially treated with all-trans-retinoic acid: a retrospective analysis of the European acute promyelocytic leukemia group. AB - PURPOSE: To retrospectively determine the outcome of acute promyelocytic leukemia (APL) patients who underwent autologous or allogeneic stem-cell transplantation (SCT) during second complete remission. PATIENTS AND METHODS: Of 122 relapsing patients included in two successive multicenter APL trials who achieved hematological second complete remission (generally after a salvage regimen of all trans-retinoic acid [ATRA] combined with chemotherapy), 73 (60%) received allogeneic (n = 23) or autologous (n = 50) SCT. RESULTS: Seven-year relapse-free survival (RFS), event-free survival (EFS), and overall survival (OS) in the autologous SCT group were 79.4%, 60.6%, and 59.8%, respectively, with a transplant-related mortality (TRM) of 6%. Of the 28 and two patients autografted with negative and positive, respectively, reverse transcriptase-polymerase chain reaction before auto SCT, three (11%) and one relapsed, respectively. In the allogeneic SCT group, 7-year RFS, EFS, and OS were 92.3%, 52.2%, and 51.8%, respectively, with 39% TRM. OS was significantly better in the autologous SCT group than in the allogeneic SCT group (P = .04), whereas RFS and EFS did not differ significantly (P = .19 and P = .11, respectively). In patients not receiving transplantation, 7-year RFS, EFS, and OS were 38%, 30.4%, and 39.5%, respectively. CONCLUSION: These retrospective data suggest that autologous SCT is very effective in APL relapsing after treatment with ATRA if performed in molecular remission. Allogeneic SCT yields few relapses, but it is associated with high TRM when performed after salvage with very intensive chemotherapy. Salvage with arsenic trioxyde, which has lower toxicity, should further improve the outcome of relapsing APL, especially before allogeneic SCT. PMID- 15534359 TI - Biology and clinical development of vascular endothelial growth factor-targeted therapy in renal cell carcinoma. AB - PURPOSE: To review the biology of renal cell carcinoma (RCC) leading to vascular endothelial growth factor (VEGF) overexpression and the clinical results of VEGF blockade in metastatic RCC. METHODS: A review of relevant published literature regarding VEGF, von Hippel-Lindau (VHL) gene inactivation and VEGF overexpression in RCC was performed. Further, a review of the mechanism, toxicity, and clinical development of VEGF-targeted therapy in metastatic RCC was undertaken. RESULTS: VEGF is the major proangiogenic protein that exerts a biologic effect through interaction with cellular receptors. The majority of sporadic clear-cell RCC tumors are characterized by VHL tumor suppressor gene inactivation. The resulting VHL gene silencing leads to VEGF overexpression. An antibody to VEGF (bevacizumab) has demonstrated a significant prolongation of time to disease progression compared with placebo in patients with metastatic RCC. Small molecules with inhibitory effects against the VEGF receptor have undergone initial clinical testing in metastatic RCC with substantial objective response rates. CONCLUSION: Therapeutic targeting of VEGF in RCC has strong biologic rationale and preliminary clinical efficacy. Further investigation will determine the optimal timing, sequence, and utility of these agents in RCC. PMID- 15534360 TI - Concomitant cisplatin significantly improves locoregional control in advanced head and neck cancers treated with hyperfractionated radiotherapy. AB - PURPOSE: To determine whether the application of two courses of cisplatin simultaneously with hyperfractionated radiotherapy improves the outcome in locally advanced and/or node-positive nonmetastatic carcinomas of the head and neck, compared with hyperfractionated radiotherapy alone. PATIENTS AND METHODS: From July 1994 to July 2000, 224 patients with squamous cell carcinomas of the head and neck (excluding nasopharynx and paranasal sinus) were randomly assigned to hyperfractionated radiotherapy (median dose, 74.4 Gy; 1.2 Gy twice daily) or the same radiotherapy combined with two cycles of concomitant cisplatin (20 mg/m2 on 5 days of weeks 1 and 5). The primary end point was time to any treatment failure; secondary end points were locoregional failure, metastatic relapse, overall survival, and late toxicity. RESULTS: There was no difference in radiotherapy between both treatment arms (74.4 Gy in 44 days). The full cisplatin dose was applied in 93% and 71% of patients during the first and second treatment cycles, respectively. Acute toxicity was similar in both arms. Median time to any treatment failure was not significantly different between treatment arms (19 months for combined treatment and 16 months for radiotherapy only, respectively) and the failure-free rate at 2.5 years was 45% and 33%, respectively. Locoregional control and distant disease-free survival were significantly improved with cisplatin (log-rank test, P = .039 and .011, respectively). The difference in overall survival did not reach significance (log-rank test, P = .147). Late toxicity was comparable in both treatment groups. CONCLUSION: The therapeutic index of hyperfractionated radiotherapy is improved by concomitant cisplatin. PMID- 15534361 TI - Concurrent radiation chemotherapy for locally advanced head and neck carcinoma: are we addressing burning subjects? PMID- 15534362 TI - A method for cloning and sequencing long palindromic DNA junctions. AB - DNA sequences containing long adjacent inverted repeats (palindromes) are inherently unstable and are associated with many types of chromosomal rearrangements. The instability associated with palindromic sequences also creates difficulties in their molecular analysis: long palindromes (>250 bp/arm) are highly unstable in Escherichia coli, and cannot be directly PCR amplified or sequenced due to their propensity to form intra-strand hairpins. Here, we show that DNA molecules containing long palindromes (>900 bp/arm) can be transformed and stably maintained in Saccharomyces cerevisiae cells lacking a functional SAE2 gene. Treatment of the palindrome-containing DNA with sodium bisulfite at high temperature results in deamination of cytosine, converting it to uracil and thus reducing the propensity to form intra-strand hairpins. The bisulfite-treated DNA can then be PCR amplified, cloned and sequenced, allowing determination of the nucleotide sequence of the junctions. Our data demonstrates that long palindromes with either no spacer (perfect) or a 2 bp spacer can be stably maintained, recovered and sequenced from sae2Delta yeast cells. Since DNA sequences from mammalian cells can be gap repaired by their co-transformation into yeast cells with an appropriate vector, the methods described in this manuscript should provide some of the necessary tools to isolate and characterize palindromic junctions from mammalian cells. PMID- 15534363 TI - A FRET-based analysis of SNPs without fluorescent probes. AB - Fluorescence resonance energy transfer (FRET) is a simple procedure for detecting specific DNA sequences, and is therefore used in many fields. However, the cost is relatively high, because FRET-based methods usually require fluorescent probes. We have designed a cost-effective way of using FRET, and developed a novel approach for the genotyping of single nucleotide polymorphisms (SNPs) and allele frequency estimation. The key feature of this method is that it uses a DNA binding fluorogenic molecule, SYBR Green I, as an energy donor for FRET. In this method, single base extension is performed with dideoxynucleotides labeled with an orange dye and a red dye in the presence of SYBR Green I. The dyes incorporated into the extended products accept energy from SYBR Green I and emit fluorescence. We have validated the method with ten SNPs, which were successfully discriminated by end-point measurements of orange and red fluorescence intensity in a microplate fluorescence reader. Using a mixture of homozygous samples, we also confirmed the potential of this method for estimation of allele frequency. Application of this strategy to large-scale studies will reduce the time and cost of genotyping a vast number of SNPs. PMID- 15534364 TI - DNA condensation and self-aggregation of Escherichia coli Dps are coupled phenomena related to the properties of the N-terminus. AB - Escherichia coli Dps (DNA-binding proteins from starved cells) is the prototype of a DNA-protecting protein family expressed by bacteria under nutritional and oxidative stress. The role of the lysine-rich and highly mobile Dps N-terminus in DNA protection has been investigated by comparing the self-aggregation and DNA condensation capacity of wild-type Dps and two N-terminal deletion mutants, DpsDelta8 and DpsDelta18, lacking two or all three lysine residues, respectively. Gel mobility and atomic force microscopy imaging showed that at pH 6.3, both wild type and DpsDelta8 self-aggregate, leading to formation of oligomers of variable size, and condense DNA with formation of large Dps-DNA complexes. Conversely, DpsDelta18 does not self-aggregate and binds DNA without causing condensation. At pH 8.2, DpsDelta8 and DpsDelta18 neither self-aggregate nor cause DNA condensation, a behavior also displayed by wild-type Dps at pH 8.7. Thus, Dps self-aggregation and Dps-driven DNA condensation are parallel phenomena that reflect the properties of the N-terminus. DNA protection against the toxic action of Fe(II) and H2O2 is not affected by the N-terminal deletions either in vitro or in vivo, in accordance with the different structural basis of this property. PMID- 15534365 TI - Cobalt hexammine induced tautomeric shift in Z-DNA: the structure of d(CGCGCA)*d(TGCGCG) in two crystal forms. AB - We report here the crystal structure of the DNA hexamer duplex d(CGCGCA)*d(TGCGCG) at 1.71 A resolution. The crystals, in orthorhombic space group, were grown in the presence of cobalt hexammine, a known inducer of the left-handed Z form of DNA. The interaction of this ion with the DNA helix results in a change of the adenine base from the common amino tautomeric form to the imino tautomer. Consequently the A:T base pair is disrupted from the normal Watson-Crick base pairing to a 'wobble' like base pairing. This change is accommodated easily within the helix, and the helical parameters are those expected for Z-DNA. When the cobalt hexammine concentration is decreased slightly in the crystallization conditions, the duplex crystallizes in a different, hexagonal space group, with two hexamer duplexes in the asymmetric unit. One of these is situated on a crystallographic 6-fold screw axis, leading to disorder. The tautomeric shift is not observed in this space group. We show that the change in inter-helix interactions that lead to the two different space groups probably arise from the small decrease in ion concentration, and consequently disordered positions for the ion. PMID- 15534366 TI - Testing the conservation of the translational machinery over evolution in diverse environments: assaying Thermus thermophilus ribosomes and initiation factors in a coupled transcription-translation system from Escherichia coli. AB - Ribosomes from the extreme thermophile Thermus thermophilus are capable of translation in a coupled transcription-translation system derived from Escherichia coli. At 45 degrees C, T.thermophilus ribosomes translate at approximately 25-30% of the maximal rate of E.coli ribosomes, and synthesize full length protein. T.thermophilus and E.coli subunits can be combined to effect translation, with the spectrum of proteins produced depending upon the source of the 30S subunit. In this system, T.thermophilus ribosomes function in concert with E.coli translational factors and tRNAs, with elongation and release factors being supplied from the E.coli extract, and purified initiation factors (IFs) being added exogenously. Cloned and purified T.thermophilus IF1, IF2 and IF3 supported the synthesis of the same products in vitro as the E.coli factors, although the relative levels of some polypeptides were factor dependent. We conclude that, at least between these two phylogenetically distant species, translational factor function and subunit-subunit interactions are conserved. This functional compatibility is remarkable given the extreme and highly divergent environments to which these species have adapted. PMID- 15534367 TI - Replication-mediated instability of the GAA triplet repeat mutation in Friedreich ataxia. AB - Friedreich ataxia is caused by the expansion of a polymorphic and unstable GAA triplet repeat in the FRDA gene, but the mechanisms for its instability are poorly understood. Replication of (GAA*TTC)n sequences (9-105 triplets) in plasmids propagated in Escherichia coli displayed length- and orientation dependent instability. There were small length variations upon replication in both orientations, but large contractions were frequently observed when GAA was the lagging strand template. DNA replication was also significantly slower in this orientation. To evaluate the physiological relevance of our findings, we analyzed peripheral leukocytes from human subjects carrying repeats of similar length (8-107 triplets). Analysis of 9400 somatic FRDA molecules using small-pool PCR revealed a similar mutational spectrum, including large contractions. The threshold length for the initiation of somatic instability in vivo was between 40 and 44 triplets, corresponding to the length of a eukaryotic Okazaki fragment. Consistent with the stabilization of premutation alleles during germline transmission, we also found that instability of somatic cells in vivo and repeats propagated in E.coli were abrogated by (GAGGAA)n hexanucleotide interruptions. Our data demonstrate that the GAA triplet repeat mutation in Friedreich ataxia is destabilized, frequently undergoing large contractions, during DNA replication. PMID- 15534368 TI - Role of CCR8 and other chemokine pathways in the migration of monocyte-derived dendritic cells to lymph nodes. AB - Studying the influence of chemokine receptors (CCRs) on monocyte fate may reveal information about which subpopulations of monocytes convert to dendritic cells (DCs) and the migration pathways that they use. First, we examined whether prominent CCRs on different monocyte subsets, CCR2 or CX3CR1, mediated migration events upstream of the accumulation of monocyte-derived DCs in lymph nodes (LNs). Monocytes were labeled and traced by uptake of latex microspheres in skin. Unexpectedly, neither CCR2 nor CX3CR1 were required. However, absence of CCR2 led to an increased labeling of the minor Gr-1int monocyte population, and the number of latex+ DCs that emigrated to LNs was correspondingly increased. Characterization of Gr-1int monocytes revealed that they selectively expressed CCR7 and CCR8 mRNA in blood. CCR7 and CCR8 pathways were used by monocyte-derived DCs during mobilization from skin to LNs. The role of CCR8 in emigration from tissues also applied to human monocyte-derived cells in a model of transendothelial trafficking. Collectively, the data suggest that Gr-1int monocytes may be most disposed to become a lymphatic-migrating DCs. When these monocyte-derived DCs exit skin to emigrate to LNs, they use not only CCR7 but also CCR8, which was not previously recognized to participate in migration to LNs. PMID- 15534369 TI - Bartonella adhesin a mediates a proangiogenic host cell response. AB - Bartonella henselae causes vasculoproliferative disorders in humans. We identified a nonfimbrial adhesin of B. henselae designated as Bartonella adhesin A (BadA). BadA is a 340-kD outer membrane protein encoded by the 9.3-kb badA gene. It has a modular structure and contains domains homologous to the Yersinia enterocolitica nonfimbrial adhesin (Yersinia adhesin A). Expression of BadA was restored in a BadA-deficient transposon mutant by complementation in trans. BadA mediates the binding of B. henselae to extracellular matrix proteins and to endothelial cells, possibly via beta1 integrins, but prevents phagocytosis. Expression of BadA is crucial for activation of hypoxia-inducible factor 1 in host cells by B. henselae and secretion of proangiogenic cytokines (e.g., vascular endothelial growth factor). BadA is immunodominant in B. henselae infected patients and rodents, indicating that it is expressed during Bartonella infections. Our results suggest that BadA, the largest characterized bacterial protein thus far, is a major pathogenicity factor of B. henselae with a potential role in the induction of vasculoproliferative disorders. PMID- 15534370 TI - Galpha13 mediates a signal that is essential for proliferation and survival of thymocyte progenitors. AB - G protein signaling via the Galpha12 family (Galpha12 and Galpha13) has not been well studied in T cells. To investigate whether Galpha12 and Galpha13 are involved in thymopoiesis, we expressed the regulator of G protein signaling domain of p115RhoGEF to inhibit Galpha12 and Galpha13 during thymopoiesis. Fetal thymus organ cultures seeded with p115DeltaDH-expressing progenitor cells showed impaired thymopoiesis with a block at the CD4-CD8-CD44-CD25+ (DN3) stage. Using Galpha13 or Galpha12 minigenes, we demonstrated that Galpha13, but not Galpha12, is required for thymopoiesis. T progenitor cells expressing p115DeltaDH showed reduced proliferation and increased cell death. T cell receptor stimulation of the fetal thymus organ cultures did not rescue the block. Overexpression of the antiapoptotic gene Bcl2 rescued the defect in DN3 cells and partially rescued T cell development. Therefore, Galpha13-mediated signaling is necessary in early thymocyte proliferation and survival. PMID- 15534372 TI - T cell-specific inactivation of the interleukin 10 gene in mice results in enhanced T cell responses but normal innate responses to lipopolysaccharide or skin irritation. AB - Interleukin (IL)-10 is a regulator of inflammatory responses and is secreted by a variety of different cell types including T cells. T regulatory cells have been shown to suppress immune responses by IL-10-dependent, but also IL-10 independent, mechanisms. Herein, we address the role of T cell-derived IL-10 in mice with an inactivation of the IL-10 gene restricted to T cells generated by Cre/loxP-mediated targeting of the IL-10 gene. Splenocytes from this T cell specific mutant secrete increased amounts of proinflammatory cytokines after activation in vitro compared with show enhanced contact hypersensitivity reactions, and succumb to severe immunopathology upon infection with Toxoplasma gondii. Despite intact IL-10 genes in other cell types, the dysregulation of T cell responses observed in the T cell-specific IL-10 mutant closely resembles the phenotype in complete IL-10 deficiency. However, in contrast to complete IL-10 deficiency, sensitivity to endotoxic shock and irritant responses of the skin are not enhanced in the T cell-specific IL-10 mutant. Our data highlight the importance of T cell-derived IL-10 in the regulation of T cell responses and demonstrate that endotoxic shock and the irritant response of the skin are controlled by IL-10 from other cell types. PMID- 15534371 TI - Number of T reg cells that differentiate does not increase upon encounter of agonist ligand on thymic epithelial cells. AB - It has been reported that the differentiation of CD4+CD25+ regulatory T cells (T reg cells) can be induced by agonist peptide/major histocompatibility complex ligands in the thymus. Exploiting a transgenic mouse line wherein expression of a particular T cell epitope can be controlled temporally and quantitatively, we found that diversion of differentiating thymocytes into the FoxP3 T reg cell pathway by this agonist ligand was essentially nonexistent. However, CD4+CD25+ thymocytes were much less sensitive than their CD4+CD25- companions, by two to three orders of magnitude, to agonist-induced clonal deletion, such that their proportion increased, giving the false impression of induced differentiation. To account for these and prior observations, one can propose that differentiation along the CD4+CD25+ pathway is induced by cues other than recognition of self agonist cues, which are poorly read by thymocytes, whose T cell receptors are conducive to selection toward the conventional CD4+CD25- lineage. Thus, selective survival, rather than induced differentiation, may explain the apparent enrichment observed here and in previous studies. PMID- 15534373 TI - Electrophysiological characterization of volume-activated chloride currents in mouse cholangiocyte cell line. AB - Recent electrophysiological and radioisotope efflux studies have demonstrated various Cl(-) channels in cholangiocytes including volume-activated Cl(-) channels (VACC). Because VACCs play prominent roles in many vital cellular functions and physiology in cholangiocytes, we have examined their electrophysiological characteristics in mouse cholangiocytes to provide an important framework for studying in the future. The present study is to characterize VACCs expressed in the mouse bile duct cell (MBDC) line, conditionally immortalized by SV40 virus. Conventional whole cell patch-clamp techniques were used to study the electrophysiological characteristics of VACC in MBDC. When the MBDCs were exposed to hypotonic solution, they exhibited an outwardly rectified current, which was significantly inhibited by replacing chloride in the bath solution with gluconate or glutamate and by administration of classic chloride channel inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoate, glybenclamide, DIDS, and tamoxifen. These inhibitory effects were reversible with washing them out from the bath solution. Moreover, the ion selectivity of the volume-activated channel to different anions indicates that it is more permeable to SCN(-) > I(-) >/= Cl(-) > F(-) >/= acetate >/= glutamate >/= gluconate. These electrophysiological characteristics demonstrate that the volume-activated current observed is a VACC. In addition, the VACC in MBDC has electrophysiological characteristics similar to those of the VACC in human cholangiocarcinoma cell line. The present study is the first to characterize the VACC in mouse cholangiocyte and will provide an important framework for further studies to examine and understand the role of the VACC in biliary secretion and ion-transport physiology. PMID- 15534374 TI - Adrenergic modulation of Escherichia coli O157:H7 adherence to the colonic mucosa. AB - Enteric neurotransmitters can modulate the biodefensive functions of the intestinal mucosa, but their role in mucosal interactions with enteropathogens is not well defined. Here we tested the hypothesis that norepinephrine (NE) modulates interactions between enterohemorrhagic Escherichia coli O157:H7 (EHEC) and the colonic epithelium. Mucosal sheets from porcine distal colon were mounted in Ussing chambers. Drugs and an inoculum of either Shiga toxin-negative or positive EHEC were added to the contraluminal and luminal bathing medium, respectively. After 90 min, adherent bacteria were quantified by an adherence assay and by immunohistochemical methods; short-circuit current (I(sc)) was measured continuously to assess changes in active ion transport. NE-treated tissues exhibited concentration-dependent increases in I(sc) and EHEC adherence. NE did not alter adherence of a rodent-adapted, noninfectious E. coli strain or two porcine-adapted non-O157 E. coli strains. The actions of NE on EHEC adherence but not I(sc) were prevented by the alpha-adrenergic antagonist yohimbine and the PKA activator Sp-8-bromoadenosine-3',5'-cyclic monophosphorothioate. Like NE, the PKA inhibitor Rp-8-bromoadenosine-3',5'-cyclic monophosphorothioate or indirectly acting sympathomimetic agents increased EHEC adherence. Nerve fibers immunoreactive for the NE-synthesizing enzymes tyrosine hydroxylase and dopamine beta-hydroxylase appeared to innervate the colonic epithelium. EHEC-like immunoreactivity on the colonic surface had the appearance of bacterial microcolonies and increased after NE treatment by a phentolamine-sensitive mechanism. Through interactions with alpha(2)-adrenergic receptors, NE appears to increase EHEC adherence to the colonic mucosa. Changes in sympathetic neural outflow may alter intestinal susceptibility to infection. PMID- 15534375 TI - Novel tools for visualizing and exploring intermolecular interactions in molecular crystals. AB - A new way of exploring packing modes and intermolecular interactions in molecular crystals is described, using Hirshfeld surfaces to partition crystal space. These molecular Hirshfeld surfaces, so named because they derive from Hirshfeld's stockholder partitioning, divide the crystal into regions where the electron distribution of a sum of spherical atoms for the molecule (the promolecule) dominates the corresponding sum over the crystal (the procrystal). These surfaces reflect intermolecular interactions in a novel visual manner, offering a previously unseen picture of molecular shape in a crystalline environment. Surface features characteristic of different types of intermolecular interactions can be identified, and such features can be revealed by colour coding distances from the surface to the nearest atom exterior or interior to the surface, or by functions of the principal surface curvatures. These simple devices provide a striking and immediate picture of the types of interactions present, and even reflect their relative strengths from molecule to molecule. A complementary two dimensional mapping is also presented, which summarizes quantitatively the types of intermolecular contacts experienced by molecules in the bulk and presents this information in a convenient colour plot. This paper describes the use of these tools in the compilation of a pictorial glossary of intermolecular interactions, using identifiable patterns of interaction between small molecules to rationalize the often complex mix of interactions displayed by large molecules. PMID- 15534376 TI - The compression mechanism of CrF3. AB - The structure of CrF3 has been studied in the pressure range from ambient to 9.12 GPa by time-of-flight neutron powder diffraction. Rietveld refinements of the crystal structure were performed in the space group R3 c for all the recorded data sets. It was found that volume reduction is achieved through rotation of the CrF6 octahedra and that the Cr-F-Cr bond angle decreases from 144.80 (7) to 133.9 (4) degrees within the investigated pressure range. Furthermore, a small octahedral strain was found to develop during compression. The octahedral strain reflects an elongation of the CrF6 octahedra along the c-axis direction. The zero pressure bulk modulus Bo and its pressure derivative Bo' were determined to be Bo = 29.2 (4) GPa and Bo' = 10.1 (3). PMID- 15534377 TI - Octahedral tilting in cation-ordered perovskites--a group-theoretical analysis. AB - Group-theoretical methods are used to enumerate the structures of ordered perovskites, in which 1:2 and 1:3 ordering of B and B' cations is considered in combination with the ubiquitous BX6 (or B'X6) octahedral tilting. The cation ordering on the B-cation site is described by irreducible representations of the Pm3 m space group of the cubic aristotype: Lambda1 (k = 1/3,1/3,1/3) for the cation ordering pattern in the 1:2 compound A3BB2'X9 and M1+ (k = 1/2,1/2,0) for the cation ordering in the 1:3 compound A4BB3'X12. The octahedral tilting is mediated by the irreducible representations M3+ and R4+. Ten distinct structures have been identified in the 1:2 case and 11 structures for 1:3. PMID- 15534378 TI - Structures of stable and metastable Ge2Sb2Te5, an intermetallic compound in GeTe Sb2Te3 pseudobinary systems. AB - The most widely used memory materials for rewritable phase-change optical disks are the GeTe-Sb2Te3 pseudobinary compounds. Among these compounds, Ge2Sb2Te5 crystallizes into a cubic close-packed structure with a six-layer period (metastable phase) in the non-thermal equilibrium state, and a trigonal structure with a nine-layer period (stable phase) in the thermal equilibrium state. The structure of the stable phase has Ge/Sb layers in which Ge and Sb are randomly occupied, as does the structure of the metastable phase, while the conventionally estimated structure had separate layers of Ge and Te. The metastable and stable phases are very similar in that Te and Ge/Sb layers stack alternately to form the crystal. The major differences between these phases are: (i) the stable phase has pairs of adjacent Te layers that are not seen in the metastable phase and (ii) only the metastable phase contains vacancies of ca 20 at. % in the Ge/Sb layers. PMID- 15534379 TI - Structure refinements of Na0.8Ti1.2Ga4.8O10: X-ray diffraction analysis for the sodium ion distribution in a one-dimensional tunnel-like space. AB - The structure of Na0.8Ti1.2Ga4.8O10 was determined by means of single-crystal X ray diffraction at 173 and 123 K and reinvestigated at 299 K. The host structure containing one-dimensional tunnels was retained over the temperatures examined, while significant changes were observed in the probability density distribution of Na+ ions in the tunnel. The refinement based on the local structure model with the deviated Na+ ion neighboring the vacancy gave a similar result to that from the conventional model, but with reduced standard uncertainties of the structural parameters for the Na+ ions. The potential barrier for the Na+ ion hopping between adjacent cavities was estimated to be ca 30-40 meV from the joint probability density function of deviated Na+ ions. PMID- 15534380 TI - Transmission electron microscopy and electron diffraction study of the short range ordering structure of alpha-LiFeO2. AB - The basic structure of alpha-LiFeO2, lithium iron oxide, is a cubic NaCl-type structure with a lattice constant of 0.42 nm; some short-range ordering characterized by octahedral clusters exists. The local structure of the short range ordering was investigated by transmission electron microscopy and electron diffraction. A new short-range ordering structure was found in local areas. The local structure has a cubic lattice with a doubled lattice constant. The occupation factors of cations on Wyckoff sites 4(a) and 4(b) are different from those on 24(d) sites, but the stoichiometric composition in cubic clusters is the same as the macroscopic composition. The number of pairs in which iron cations exist in nearest-neighbor sites and next nearest-neighbor sites is reduced in the structure. This means that a magnetic interaction between the iron cations is reduced by cation ordering even without spin ordering at room temperature. PMID- 15534381 TI - Phase transitions in K2Cr2O7 and structural redeterminations of phase II. AB - Crystals of phase II K2Cr2O7, potassium dichromate, space group P1 , grown from aqueous solution undergo a first-order transition to phase I, space group reportedly P21/n, at a phase-transition temperature, TPT, of 544 (2) K on first heating; the corresponding transition on cooling is at 502 (2) K. The endotherm on subsequent heatings occurs reproducibly at TPT = 531 (2) K. Mass loss between ca 531 and 544 K, identified as included water, is rapid and continues more slowly to higher temperatures for a total loss of ca 0.20%. The higher TPT on first heating is associated with the increasing pressure of superheated water occupying inclusion defects. The latent diagonal glide plane in phase II allows the structure of phase I to be inferred. The triclinic structure at 296 K has been independently redetermined. Normal probability analysis shows high consistency between the resulting and previous atomic coordinates, but with uncertainties reduced by a factor of ca 2. The earlier uncertainties are systematically underestimated by a comparable factor. The structure of phase IIb, space group A2/a on transposing axes, was determined at ca 300 K by Krivovichev et al. [Acta Cryst. (2000), C56, 629-630]. The first-order transition between phases I and II arises from the ca 60 degrees relative rotation of terminal O atoms in each tetrahedron as the n glide plane is gained or lost. A transition between phases IIb and I, also of first order, is likely but not between phases II and IIb. An intermediate phase may exist between phases IIb and I. PMID- 15534382 TI - Structural chemistry of new lithium bis(oxalato)borate solvates. AB - Recently lithium bis(oxalato)borate, LiB(C2O4)2, has been proposed as an alternative lithium salt for the electrolyte in rechargeable batteries that do not contain explosive perchlorate, reactive fluoride or toxic arsenic. This lithium salt crystallizes in the form of solvates from such solvents as water, acetonitrile, acetone, dimethoxyethane, 1,3-dioxolane and ethylene carbonate. Their crystal structures were determined in order to explore the crystal chemistry of this lithium salt. It was found that most of the solvents consist of a lithium bis(oxalato)borate dimer in which the ligand acts as both a chelating and a bridging agent. Lithium has octahedral coordination that typically includes one or, less commonly, two solvent molecules. An exception to this rule is the ethylene carbonate solvate where the lithium is tetrahedrally surrounded exclusively by the solvent and bis(oxalato)borate plays the role of counter-ion only. The ethylene carbonate solvates were also studied for LiPF6 and LiAsF6 salts and they have similar structures to the bis(oxalato)borate tetrahedral complexes. PMID- 15534383 TI - Parameterization of the close packing of molecules in the unit cell. AB - The box model of crystal packing describes unit cells in terms of a limited number of arrangements, or packing patterns, of molecular building blocks. Cell dimensions have been shown to relate to molecular dimensions in a systematic way. The distributions of pattern coefficients (cell length/molecular dimension) for thousands of structures belonging to P21/c, P1 , P212121, P21 and C2/c are presented and are shown to be entirely consistent with the box model of crystal packing. Contributions to the form of the histograms from molecular orientation and molecular overlap are discussed. Gaussian fitting of the histograms has led to the parameterization of close packing within the unit cell and it is shown that molecular crystal structures are very similar to one another at a fundamental level. PMID- 15534384 TI - Di-p-bromophenyl ether, a redetermined crystal structure derived from low-quality diffraction data. AB - We show that the lack of good quality data, normally essential to successful crystal structure analysis, can in part be compensated for by measuring data from several crystals and merging the resulting data sets. The crystal structure of the flame retardant di-p-bromophenyl ether, C12H8Br2O, a twofold axially symmetric molecule, has been redetermined and refined from such a merged multi crystal diffraction data set to an acceptable conventional R factor (R1 = 0.06), a result which could not have been obtained from any one of our single-crystal diffraction data sets used alone in the normal manner. PMID- 15534385 TI - Structural investigations of phosphorus-nitrogen compounds. 6. Relationships between molecular parameters in per-X-substituted bridged spermine derivatives and basicity constants SigmaalphaR of substituents. AB - A systematic study is reported of the products of the nucleophilic substitution reactions of the spermine-bridged cyclotriphosphazene, [N3P3X4(NHCH2CH2CH2N)CH2CH2]2 [where X = Cl (2a)], to give a number of new structures [(2b)-(2g)] in which X = OPh, [spiro-O(CH2)3O]0.5, Ph, NHPh, NC4H8 and NHBut, respectively. A comparison has been made between the sum of the substituent basicity constants, Sigmaalpha(R), obtained in nitrobenzene solution, and ten molecular parameters of the N3P3 ring (the internal bond angles alpha, beta, gamma, delta and theta;, and the P-N bond lengths a, b, c, d and e) as well as the difference between the bond lengths a and b, Delta(P-N). It is found that the systematic change in molecular parameters of compounds (2a)-(2g) is in line with changes in alphaR values, indicating the similarity in relative electron releasing capacity of substituents X in the solid state and in solution. It is also found that the effect on molecular parameters of (2a)-(2g) with two X substituents in PX2 groups is greater than that for one X substituent in P(OR)X groups in an analogous series of compounds observed previously [Besli et al. (2002). Acta Cryst. B58, 1067-1073]. PMID- 15534386 TI - Geometry of the 2-aminoheterocyclic-carboxylic acid R2(2)(8) graph set: implications for crystal engineering. AB - The geometry of the R2(2)(8) graph set formed between a 2-aminoheterocyclic ring containing an Nsp2 atom (in the 1-position of the ring) and a carboxylic acid has been studied. Collating data from known co-crystal structures containing five- and six-membered heterocyclic rings from the Cambridge Structural Database revealed unexpected differences between two kinds of non-hydrogen contact distances, and between specific bond distances and angles of the heterocycle. Not only were the interatomic non-hydrogen distances between the N atoms (heterocycle) and O atoms (carboxylate) asymmetric, but also the 2-amino N atom (N21) to the heterocyclic C atom (C2) bond was shorter than the C2 to N1sp2 bond. However, this shortening of the C2-N21 bond was not observed in the examples where N21 was substituted with a non-H atom. For the six-membered rings the data also showed that as the C2-N21 bond shortened the N1-C2-N21 bond angle increased. PMID- 15534387 TI - Different forms of antiparallel stacking of hydrogen-bonded antidromic rings in the solid state: polymorphism with virtually the same unit cell and two dimensional isostructurality with alternating layers. AB - As a continuation of a systematic structural analysis of 2 hydroxycycloalkanecarboxylic acids and their carboxamide analogs, the effects of antidromic rings [Jeffrey & Saenger (1991). Hydrogen Bonding in Biological Structures. Berlin, Heidelberg: Springer Verlag] upon the layer stacking of cyclopentane and cycloheptane derivatives are compared. Determination of the structure of trans-2-hydroxycycloheptanecarboxylic acid (2) led to the discovery of two polymorphs with virtually the same unit cell [Kalman et al. (2003). J. Am. Chem. Soc. 125, 34-35]. (i) The layer stacking of the antidromic rings for the whole single crystal is antiparallel (2b). (ii) The antidromic rings and the 21 axis are parallel (2a), consequently the domains of the single crystal must be antiparallel. While their polymorphism is solvent-controlled, they illustrate a novel form of two-dimensional isostructurality. Antiparallel layer stacking is again demonstrated by trans-2-hydroxycycloheptanecarboxamide (3) (space group Pbca). It is built up from layers isostructural with those in the homologous trans-2-hydroxycyclopentanecarboxamide (4) [Kalman et al. (2001). Acta Cryst. B57, 539-550], but in this structure (space group Pca21) the layers are stacked in parallel mode. Similar to (2a) and (2b), the antiparallel layer stacking in (3) versus their parallel array in (4) illustrates the two-dimensional isostructurality with alternating layer orientations. Although (3) and (4) display isostructurality, they are not isomorphous. PMID- 15534389 TI - 2-[(E)-(4-Chlorophenyl)methyleneamino]-N-(X-methylphenyl)-4,5,6,7-tetrahydro-1 benzothiophene-3-carboxamide, where X = 2 and 3. AB - The title compounds, both C(23)H(21)ClN(2)OS, are isomeric, with (I) and (II) being the N-3-methylphenyl and N-2-methylphenyl derivatives, respectively. The dihedral angle between the 4-chlorophenyl group and the thiophene ring in (II) [38.1 (1) degrees ] is larger than that in (I) [7.1 (1) degrees ], indicating steric repulsion between the chlorophenyl and o-toluidine groups in (II). In both compounds, an intramolecular N-H...N hydrogen bond forms a pseudo-six-membered ring, thus locking the molecular conformation. In the crystal structures, molecules are connected via N-H...O hydrogen bonds, forming chains along the b axis in (I) and along the c axis in (II). Intermolecular C-H...O/S and pi-pi interactions are also observed in (II), but not in (I). PMID- 15534390 TI - Analysis of the secondary structure of the cys-less yeast mitochondrial citrate transport protein and four single-cys variants by circular dichroism. AB - Utilizing cysteine scanning mutagenesis, with functional Cys-less citrate transport protein (CTP) serving as the starting template, we previously demonstrated that four single-Cys mutants located in transmembrane domains III and IV, rendered the CTP nonfunctional. The present investigations assess and quantify the secondary structure of the Cys-less CTP and the four single-Cys mutants, both in the absence and presence of citrate, via circular dichroism (CD) spectroscopy. In detergent micelles, highly purified Cys-less CTP contained approximately 50% alpha-helix and approximately 20% beta-sheet. The CD spectra of the G119C, E122C, R181C, and R189C mutants in detergent micelles were virtually superimposable with that of the functional Cys-less CTP, thereby suggesting that the wild-type residues, rather than affecting structure, may assume important mechanistic roles. Exogenously added citrate caused a significant change in the CD spectra of all solubilized CTP samples. Analyses of the spectra of the Cys less CTP indicated an approximately 10% increase in its alpha-helical content in the presence of citrate. The conformational changes effected by the addition of substrate were less pronounced with the single-Cys mutants. Studies of the Cys less CTP reconstituted in liposomes indicated that while the CD spectra was red shifted, the net secondary structure of the reconstituted carrier is approximately equivalent to that of the transporter in detergent micelles, and displayed a response to added citrate. In combination, the above studies indicate that purified Cys-less CTP in either sarkosyl micelles or in liposomes, and the four inactive single-Cys mutants in sarkosyl micelles, retain native-like structure, and thus represent ideal material for detailed structural characterization. PMID- 15534391 TI - Different subunit location of the inhibition and transport sites in the mitochondrial calcium uniporter. AB - The mitochondrial calcium uniporter behaves as a cooperative mechanism, where the velocity is dependent on [Ca2+]ex. Transport kinetics follows a sigmoidal behavior with a Hill coefficient near 2.0, indicating the binding of at least two calcium molecules. Calcium transport in mitochondria is dependent on a negative inner membrane potential and is inhibited by policationic ruthenium compounds. In this study, calcium uptake activity was reconstituted into cytochrome oxidase vesicles by incorporating solubilized mitochondrial proteins. Calcium accumulation plotted against increasing Ca2+ concentrations followed a sigmoidal behavior with a Hill coefficient of 1.53. The uptake was sensitive to ruthenium policationic inhibitors, e.g. ruthenium red and Ru360. After mitochondrial proteins were separated by preparative isoelectrofocusing and incorporated into cytochrome oxidase vesicles, two peaks of calcium uptake activity were recovered. One of the activities was inhibited by Ru360, while the second activity was insensitive to Ru360 and was associated with proteins focused at very acidic isoelectric points. By using a thiol-group crosslinker and radiolabeled Ru360, we proposed a scheme of partial dissociation of the uniporter inhibitor-binding subunit under acidic conditions. PMID- 15534392 TI - Inhibitory and anchoring domains in the ATPase inhibitor protein IF1 of bovine heart mitochondrial ATP synthase. AB - The inhibitor protein IF1 is a basic protein of 84 residues which inhibits the ATPase activity of the mitochondrial FoF1-ATP synthase complex without having any effect on ATP synthesis. Results of cross-linking and limited proteolysis experiments are presented showing that in the intact FoF1 complex "in situ," in the inner membrane of bovine heart mitochondria, the central segment of IF1 (residues 42-58) binds to the alpha and beta subunits of F1 in a pH dependent process, and inhibits the ATPase activity. The C-terminal region of IF1 binds, simultaneously, to the OSCP subunit of Fo in a pH-independent process. This binding keeps IF1 anchored to the complex, both under inhibitory conditions, at acidic pH, and noninhibitory conditions at alkaline pH. PMID- 15534393 TI - The alternative respiratory pathway of euglena mitochondria. AB - Mitochondria, isolated from heterotrophic Euglena gracilis , have cyanide resistant alternative oxidase (AOX) in their respiratory chain. Cells cultured under a variety of oxidative stress conditions (exposure to cyanide, cold, or H2O2) increased the AOX capacity in mitochondria and cells, although it was significant only under cold stress; AOX sensitivity to inhibitors was also increased by cold and cyanide stress. The value of AOX maximal activity reached 50% of total respiration below 20 degrees C, whereas AOX full activity was only 10-30% of total respiration above 20 degrees C. The optimum pH for AOX activity was 6.5 and for the cytochrome pathway was 7.3. GMP, AMP, pyruvate, or DTT did not alter AOX activity. The reduction level of the quinone pool was higher in mitochondria from cold-stressed than from control cells; furthermore, the content of reduced glutathione was lower in cold-stressed cells. Growth in the presence of an AOX inhibitor was not affected in control cells, whereas in cold-stressed cells, growth was diminished by 50%. Cyanide diminished growth in control cells by 50%, but in cold-stressed cells this inhibitor was ineffective. The data suggest that AOX activity is part of the cellular response to oxidative stress in Euglena . PMID- 15534394 TI - Mitochondrial respiratory chain complex patterns from Acanthamoeba castellanii and Lycopersicon esculentum: comparative analysis by BN-PAGE and evidence of protein-protein interaction between alternative oxidase and complex III. AB - We have previously shown that a kinetic interplay exists between the cytochrome pathway and the alternative oxidase in mitochondria from amoeba Acanthamoeba castellanii . Native interaction analyses using blue native gel electrophoresis coupled to denaturating electrophoresis and immunodetection have indicated associations between alternative oxidase and oxidative phosphorylation complexes in both amoeba and tomato mitochondria. These associations are dependent on the expression level of alternative oxidase according to the physiological state in both organisms. Alternative oxidase associates broadly with large complexes of the respiratory chain when it is expressed in large amount, i.e., in ripe tomato and exponentially growing amoeba. On the contrary, alternative oxidase interacts specifically with complex III even if expression of the oxidase is low, i.e., in green tomato and stationary phase amoeba. This specific interaction represents a higher level of regulation driven by protein-protein interactions leading to a direct kinetic interplay between the cytochrome pathway and alternative oxidase in both plant and amoeba mitochondria. PMID- 15534395 TI - A transplasma membrane redox system in Phycomyces blakesleeanus: properties of a ferricyanide reductase activity regulated by iron level and vitamin K3. AB - Intact Phycomyces blaskesleeanus mycelia are capable of reducing extracellular ferricyanide and this transmembrane reduction is an enzymatic process, which is enhanced by the presence of 10 mM lactate. It is modulated in response to intracellular iron levels and negatively regulated by iron and copper. It is inhibited by NEM, p CMB, iodoacetate, Zn2+, Cd2+, dicumarol, and capsaicine analog, but not by cloroquine, and activated by Ca2+, Mg2+, Na+, and K+. Ferricyanide reduction was concomitant with proton release into the extracellular medium, both processes being greatly promoted by vitamin K3 following hyperbolic saturation kinetics with regard to ferricyanide concentration. No stoichiometric proton release was observed with regard to ferricyanide reduction in the absence or the presence of vitamin K3. Proton release coupled with ferricyanide reductase activity does not appear to be due to H+-ATPase. The relevance of these findings to the relationship between the two processes is discussed. PMID- 15534396 TI - Redox state of endogenous coenzyme q modulates the inhibition of linoleic acid induced uncoupling by guanosine triphosphate in isolated skeletal muscle mitochondria. AB - The skeletal muscle mitochondria contain two isoforms of uncoupling protein, UCP2 and mainly UCP3, which had been shown to be activated by free fatty acids and inhibited by purine nucleotides in reconstituted systems. On the contrary in isolated mitochondria, the protonophoretic action of muscle UCPs had failed to be demonstrated in the absence of superoxide production. We showed here for the first time that muscle UCPs were activated in state 3 respiration by linoleic acid and dissipated energy from oxidative phosphorylation by decreasing the ADP/O ratio. The efficiency of UCPs in mitochondrial uncoupling increased when the state 3 respiratory rate decreased. The inhibition of the linoleic acid-induced uncoupling by a purine nucleotide (GTP), was not observed in state 4 respiration, in uninhibited state 3 respiration, as well as in state 3 respiration inhibited by complex III inhibitors. On the contrary, the progressive inhibition of state 3 respiration by n -butyl malonate, which inhibits the uptake of succinate, led to a full inhibitory effect of GTP. Therefore, as the inhibitory effect of GTP was observed only when the reduced state of coenzyme Q was decreased, we propose that the coenzyme Q redox state could be a metabolic sensor that modulates the purine nucleotide inhibition of FFA-activated UCPs in muscle mitochondria. PMID- 15534397 TI - Quality of primary care guidelines for acute low back pain. AB - STUDY DESIGN: Systematic review of clinical guidelines. OBJECTIVES: To assess the methodologic quality of existing guidelines for the management of acute low back pain. SUMMARY OF BACKGROUND DATA: Guidelines are playing an increasingly important role in evidence-based practice. After publication of the Quebec Task Force in Canada in 1987 and the Agency for Health Care Policy and Research guidelines in the United States in 1994, guidelines for acute low back pain were developed in many other countries. However, little is known about the methodologic quality of these guidelines. METHODS: Guidelines were selected by electronically searching MEDLINE and the Internet and through personal communication with experts in the field of low back pain research in primary care. The methodologic quality of the guidelines was assessed by two authors independently using the AGREE instrument. RESULTS: A total of 17 guidelines were included. Overall, the quality of reporting of guidelines was disappointing. Most guidelines clearly described the aim of the guideline and its target population, and most guideline development committees were multiprofessional. However, many other methodologic flaws were identified. More than half of the guidelines did not take patients' preferences into account, did not perform a pilot test among target users, did not clearly describe the methods of study identification and selection, did not include an external review, did not provide a procedure for updating, were not supported with tools for application, did not consider potential organizational barriers and cost implications, did not provide criteria for monitoring and audit, did not include recommendations for implementation strategies, and did not adequately record editorial independence and conflict of interest of the members. The diagnostic and therapeutic recommendations of the guidelines were largely similar. CONCLUSIONS: The quality and transparency of the development process and the consistency in the reporting of primary care guidelines for low back pain need to be improved. PMID- 15534398 TI - The transpedicular approach in the management of thoracic spine tuberculosis: a short-term follow up study. AB - STUDY DESIGN: A retrospective descriptive assessment of the clinical and radiologic outcomes of 11 patients who underwent transpedicular decompression for thoracic spine tuberculosis. OBJECTIVES: To study the neurologic and radiologic outcomes in patients who underwent transpedicular decompression for thoracic spine tuberculosis. SUMMARY OF BACKGROUND DATA: Several approaches have been used in the management of thoracic spine tuberculosis to achieve the goals of decompression of the cord followed by immobilization and antituberculous therapy. These range from conservative regimens of computed tomography-guided biopsy followed by bed rest and drug therapy to radical surgeries that involve extensive debridement of the vertebral body followed by instrumentation. The authors report their experience with a "middle path" regimen of transpedicular decompression followed by external immobilization and antituberculous therapy. METHODS: The charts of 11 patients were reviewed retrospectively for clinical outcome, and kyphotic angle was measured on the follow-up radiographs to ascertain progression of kyphosis. RESULTS: There was no worsening of the neurologic status in any patient, and 10 of the 11 patients returned to functional activity. There was no significant progression of kyphosis. CONCLUSIONS: Our results show that the transpedicular approach is a viable and safe surgical option for ventral decompression in thoracic spine tuberculosis, followed by chemotherapy for 18 months and immobilization in an alkathene shell for 3 months. PMID- 15534399 TI - Cervical spine metastasis of multiple myeloma: a case report with 16 years of follow-up. AB - STUDY DESIGN: Presented is a case report of a 16-year clinical course of a patient with a multiple myeloma of the third cervical vertebra. OBJECTIVE: To describe a rare 16-year follow-up of a cervical multiple myeloma lesion that forced several surgical interventions. SUMMARY OF BACKGROUND DATA: Multiple myeloma is a systemic malignancy. It slowly causes bone destruction due to bone marrow infiltration. It is more common in the elderly and rarely affects the young. In patients with continuing neck pain and headache, one has to bear in mind the rare possibility of multiple myeloma or also other malignancies affecting the cervical spine. MATERIALS AND METHODS: Clinical case analysis, radiographs, magnetic resonance images, and histological sections of the lesion are discussed. Repeated surgical interventions were required due to progressive disease. RESULTS: Initial surgical treatment was aimed at palliation. However, this was in contrast to the slowly progressive clinical course. Thus, further surgical interventions were necessary. The patient died 16 years after the first procedure due to progression of the disease. CONCLUSION: Even in young patients presenting with neck pain and minor neurological symptoms, one should bear in mind the rare possibility of multiple myeloma. At the very least, a radiograph of the cervical spine and a routine blood test should be performed. Surgical interventions must take into account the relatively long period of survival in multiple myeloma patients compared to patients with other secondary bone tumors. PMID- 15534400 TI - Thoracic spondylitis from a mycotic (Streptococcus pneumoniae) aortic aneurysm: a case report. AB - STUDY DESIGN: We report on a 54-year-old man with chronic lower back pain after recent streptococcus pneumoniae pulmonary infection, resulting in a mycotic aortic aneurysm and spondylodiscitis of the eighth vertebrae 6 months later. Successful surgical treatment and recurrence-free survival after 4 years are described. SUMMARY OF BACKGROUND DATA: Osteomyelitis by Streptococcus pneumoniae of the spine combined with contained rupture of a mycotic aortic aneurysm into lung and spine has not been reported to date. Mycotic aneurysms with pulmonary fistulas are reported to carry a mortality rate of up to 100%. Few cases have been reported with different operative and conservative strategies. METHODS: The mycotic aortic aneurysm was excised using extracorporeal circulation and replaced by a Dacron graft. The spondylitic section of the eighth thoracic vertebrae was radically resected, and a tricortical bone block from the iliac crest was inserted into the defect. To keep compartments separated, collagen sponges with antibiotic supplementation were used. A triple antibiotic therapy (Metronidazol 3 x 0.5 g/day, Cefotaxim 3 x 2 g/day, and Flucloxacillin 3 x 2 g/day) was prescribed for 6 weeks and changed to Clindamycin for 1 year thereafter. RESULTS: The patient made a good recovery and is free of recurrence 4 years after surgery. CONCLUSIONS: Lower back pain might be a projected pain. Particularly in older patients or in the presence of comorbidities resulting in an immunocompromised status, an aggressive workup may be indicated. Radical resection of inflammatory tissues, sparse use of implant material, and prolonged administration of antibiotics proved a successful strategy in this patient. PMID- 15534401 TI - Solid variant of aneurysmal bone cyst of the cervical spine. AB - STUDY DESIGN: A case of the solid variant of aneurysmal bone cyst affecting the posterior component of the fourth cervical vertebra is reported. Imaging studies showed an expansile destructive lesion. After curettage, autologous iliac bone grafting with posterior fusion was performed. There was no sign of local recurrence 2 years after surgery. OBJECTIVES: To emphasize the occurrence of the solid variant of aneurysmal bone cyst in the cervical spine. SUMMARY OF BACKGROUND DATA: The solid variant of aneurysmal bone cyst is rare, and only 12 cases occurring in the vertebrae, including 3 in the cervical vertebrae, have been reported. The condition is difficult to diagnose radiologically before biopsy or surgery. METHODS: A 9-year-old girl presented with pain in the nape of the neck without any neurologic deficit. She was found to have the solid variant of aneurysmal bone cyst in the posterior component of the fourth cervical vertebra, which had destroyed the lamina and spinous process. Part of the posterior aspect of the C4 vertebral body was also involved. Curettage of the lesion was performed, and the defect in the posterior component of the vertebra was reconstructed using an autologous iliac bone graft with posterior fusion using a halo vest. RESULTS: Magnetic resonance imaging disclosed a homogeneous low intensity mass at the lamina, spinous process, and vertebral body of C4 on T1 weighted images. The mass showed heterogeneous high signal intensity on Gd enhanced images. Histologically, the resected specimen showed predominant fibroblastic proliferation, with minor foci of reactive osteoid formation and an area of osteoclast-like giant cells. Neither cellular atypia nor mitotic figures were evident. There was no sign of local recurrence 2 years after surgery. CONCLUSIONS: The solid variant of aneurysmal bone cyst should be included in the differential diagnosis of any lytic expansile lesion of the spine, even though it is a destructive lesion. Gd-enhanced magnetic resonance imaging may be helpful for distinguishing the solid variant from conventional aneurysmal bone cyst. PMID- 15534403 TI - Bioabsorbable interbody cages in a sheep cervical spine fusion model. AB - STUDY DESIGN: An experimental study using a sheep cervical spine interbody fusion model. OBJECTIVES: To compare interbody fusion of an autologous tricortical iliac crest bone graft with two bioabsorbable cages and to determine whether there are differences between the three interbody fusion techniques in 1) the ability to preserve postoperative distraction, 2) the biomechanical stability, and 3) the histologic characteristics of intervertebral bone matrix formation. SUMMARY AND BACKGROUND DATA: Bioabsorbable cages would be beneficial compared with metallic cages; however, currently no suitable bioabsorbable interbody fusion cage is available. METHOD: Twenty-four sheep underwent C3/C4 discectomy and fusion. The following stabilization techniques were used: Group 1) autologous tricortical iliac crest bone graft (n = 8); Group 2) bioabsorbable cage made of 70/30 poly(l lactide-co-d,l-lactide) (experimental) filled with autologous cancellous bone graft (n = 8); Group 3) bioabsorbable cage made of a polymer-calciumphosphate composite (Biomet Europe, Dordrecht, The Netherlands) filled with autologous cancellous bone graft (n = 8). Radiographic scans to determine disc space height were performed before and after surgery and after 1, 2, 4, 8, and 12 weeks, respectively. After 12 weeks, animals were killed, and fusion sites were evaluated using functional radiographic views in flexion and extension. Quantitative computed tomographic scans were used to assess bone mineral density, bone mineral content, and bony callus volume. Biomechanical testing was performed in flexion, extension, axial rotation, and lateral bending to determine stiffness, ROM, and neutral and elastic zone. Histomorphological and histomorphometrical analysis were performed to evaluate fusion and foreign body reactions associated with the bioabsorbable cages. RESULTS: Over a 12-week period, the polymer-calciumphosphate composite cage showed significantly higher values for disc space height compared with the bone graft and the poly(l-lactide co-d,l-lactide) cage. Additionally, the polymer-calciumphosphate composite cage demonstrated a significantly higher stiffness and lower ROM, neutral zone, and elastic zone in axial rotation and lateral bending than any other group. However, quantitative computed tomographic scans demonstrated cracks in six of the eight polymer-calciumphosphate composite cages after 12 weeks. Histologically, the highest bone volume/total volume ratio and the highest fusion rate were found in the polymer-calciumphosphate composite cage group. Although the poly(l-lactide-co d,l-lactide) cage showed grade I through III foreign body reactions in all fusion areas, only two animals developed grade I foreign body reactions with the polymer calciumphosphate composite cage. CONCLUSION: After 12 weeks, there was no significant difference between the bioabsorbable poly(l-lactide-co-d,l-lactide) cage and the tricortical bone graft. In comparison to the tricortical bone graft, the bioabsorbable polymer-calciumphosphate composite cage showed significantly better distractive properties, a significantly higher biomechanical stiffness, and an advanced interbody fusion; however, six of eight polymer-calciumphosphate composite cages cracked. Although the fate of the foreign body reactions and the cracks is currently unclear for both bioabsorbable cages, the early appearance of large osteolysis associated with use of the poly(l-lactide-co-d,l-lactide) cage allows skepticism regarding the value of this bioabsorbable implant. PMID- 15534405 TI - Infliximab attenuates immunoreactivity of brain-derived neurotrophic factor in a rat model of herniated nucleus pulposus. AB - STUDY DESIGN: The effect of infliximab, a chimeric monoclonal antibody to TNF alpha, on induction of brain-derived neurotrophic factor (BDNF) was examined using an experimental herniated nucleus pulposus (NP) model. OBJECTIVES: To investigate whether treatment of infliximab could attenuate an induction of BDNF, which functions as a modulator of pain, following NP application to the nerve root. SUMMARY OF BACKGROUND DATA: Evidence from basic scientific studies proposes that TNF-alpha is involved in the development of NP-induced nerve injuries. However, the therapeutic mechanisms of infliximab against pain have not been elucidated experimentally. METHODS: Twenty rats were used in this study. In the test groups, the animals underwent application of NP to the L4 nerve roots and received a single systemic (intraperitoneal) injection of infliximab at the time of surgery (Infli-0 group, n = 5) or at 1 day after operation (Infli-1 group, n = 5). As a control treatment, sterile water was administered intraperitoneally to 5 rats with NP application (NP group) and to 5 sham-operated rats (sham group). On day 3 after surgery, the L4 dorsal root ganglion (DRG) and L4 spinal segment were harvested and assessed regarding BDNF immunoreactivity. RESULTS.: Application of NP induced a marked increase of BDNF immunoreactivity in number in the DRG neurons and within the superficial layer in the dorsal horn compared with the sham group (P < 0.01). Infliximab treatment in the Infli-0 and Infli-1 groups reduced the BDNF induction in both DRG and spinal cord (P < 0.05). CONCLUSION: These findings indicate that infliximab attenuates the elevated BDNF levels induced by NP. The present study therefore further indicates the importance of TNF-alpha in sciatica due to disc herniation and the possible therapeutic use of a TNF-alpha inhibitor for this condition. PMID- 15534406 TI - Cerebrospinal fluid biomarkers in experimental spinal nerve root injury. AB - STUDY DESIGN: Cerebrospinal fluid biomarkers were evaluated in a setup using established pig models to mimic clinical disc herniation. OBJECTIVES: To investigate biomarkers for nerve tissue injury, inflammation, and pain in cerebrospinal fluid after mechanical compression and/or nucleus pulposus application to spinal nerve roots. SUMMARY OF BACKGROUND DATA: The association between mechanical compression, biochemical effects of nucleus pulposus, and nerve root injury in degenerative disc disorders is incompletely investigated. METHODS: The unilateral S1 nerve root was exposed in 20 pigs. The animals were divided into four groups (n = 5 each): 1) slow-onset mechanical compression with an ameroid constrictor; 2) autologous nucleus pulposus application; 3) mechanical compression plus nucleus pulposus; and 4) sham operation. After 1 week, 6 mL of cerebrospinal fluid was collected, and four structural nerve proteins, neurofilaments, S-100, glial fibrillary acidic protein, neuron-specific enolase, the proinflammatory cytokine interleukin-8, the neurotransmitter nociceptin, and substance P endopeptidase activity were analyzed using immunoassays. RESULTS: The concentration of neurofilament was increased in the mechanical compression group (17.0 microg/L +/- 5.0) and in the mechanical compression plus nucleus pulposus group (19.8 +/- 12.1 microg/L) compared with the sham group (0.9 +/- 0.9 microg/L) and the nucleus pulposus group (0.4 +/- 0.1 microg/L) (P < 0.01 for both). The concentration of nociceptin was increased significantly in the mechanical compression group (24.0 +/- 8.6 fm/mL) and in the mechanical compression plus nucleus pulposus group (31.2 +/- 6.6 fm/mL) compared with the sham group (7.0 +/- 1.3 fm/mL) (P < 0.05 and P < 0.01, respectively). A correlation was found between concentrations of neurofilament and nociceptin (r = 0.50, P < 0.05). There were no intergroup differences regarding glial fibrillary acidic protein, neuron-specific enolase, S-100, interleukin-8, or substance P endopeptidase activity. CONCLUSIONS: The present study demonstrates increased concentrations of neurofilament and nociceptin in cerebrospinal fluid after nerve root compression. A simultaneous application of nucleus pulposus did not increase the response. PMID- 15534407 TI - Biomechanical analysis of transpedicular screw fixation in the subaxial cervical spine. AB - STUDY DESIGN: An in vitro biomechanical study to compare 2 different dorsal screw fixation techniques in the cervical spine with respect to primary stability and stability after cyclic loading. OBJECTIVES: To investigate if the biomechanical stability is better in pedicle screw or in lateral mass fixation. SUMMARY OF BACKGROUND DATA: In patients with poor bone quality who require multisegmental fixations, the current dorsal stabilization procedures in the subaxial cervical spine using lateral mass screws are often insufficient. Cervical pedicle screw fixation has been suggested as an alternative procedure, but there are still limited data available on the biomechanical differences between pedicle screw and lateral mass fixation. METHODS: A severe multilevel discoligamentous instability was created in 8 human cervical spine specimens (C2-C7). Dorsal stabilization was performed with the assistance of computer navigation (SurgiGate, Medivison, Switzerland) using either lateral mass or pedicle screw fixation. In the first part of the study, primary stability was measured by means of a multidirectional flexibility test. Then, specimens were divided into 2 groups, randomized for bone mineral density. Cyclic loading was applied with sinusoidal loads in flexion/extension (1000 cycles, +/-1.5 Nm, 0.1 Hz). Mechanical behavior of the specimens was determined by a flexibility test before and after the application of cyclic loads. Data analysis was performed by calculating the ranges of motion, and statistical differences were determined with the t test for group comparison. RESULTS: Pedicle screw fixation showed a significantly higher stability in lateral bending (pedicle screw range of motion 0.86 +/- 0.31 degrees; lateral mass range of motion 1.43 +/- 0.62 degrees; P = 0.037). No significant differences were seen in flexion/extension and axial rotation. After cyclic loading, the decrease in stability was less with pedicle screw fixation in all load directions. Differences in the decrease of stability were statistically significant in flexion/extension (pedicle screw 95.4 +/- 9.4%; lateral mass 70.5 +/- 9.8%; P = 0.010) and lateral bending (pedicle screw 105.3 +/- 5.0%; lateral mass 84.2 +/- 13.6%; P = 0.046), whereas there was no significant difference in axial rotation. CONCLUSIONS: The major finding of the current study was the higher stability of pedicle screws over lateral mass fixation with respect to primary stability and stability after cyclic loading. From a biomechanical point of view the use of pedicle screws in the subaxial cervical spine seems justified in patients with poor bone quality and need for multisegmental fixation. PMID- 15534408 TI - Morphometric evaluation of subaxial cervical vertebrae for surgical application of transpedicular screw fixation. AB - STUDY DESIGN: A morphometric evaluation of pedicle projections in 29 cervical spinal columns (C3-C7) for three-dimensional surgical anatomy for safe surgery was proposed. OBJECTIVE: In this study, pedicles and intimate structures of the subaxial vertebrae from C3 to C7 were evaluated to provide some morphometric data for cervical transpedicular screw fixation. SUMMARY OF BACKGROUND DATA: Detailed knowledge of surgical anatomy and variation of the subaxial vertebrae is a must for safe and effective surgery of the region. Although there are several clinical studies of transpedicular fixation, few studies have been performed on cervical pedicle measurements and their projection. METHODS: In 29 dried bone cervical spinal columns (C3-C7), pedicle dimensions (pedicle height, width, length), measurements of lateral mass and pedicle length distance and pedicle axis length, investigation of distances of superior facet-midpedicle axis and inferior facet midpedicle axis, and transverse and sagittal angles of the pedicles were performed in linear and angular measurements. RESULTS: The obtained data from the series revealed that the mean values were approximately ranging from 6.7 to 7.2 mm for pedicle height, 4.4 to 4.9 mm for pedicle width, 5.3 to 6.2 mm for pedicle length, 15.3 to 16.0 mm for lateral mass and pedicle length, 22.2 to 27.7 mm for pedicle axis length, 3.8 to 5.3 mm for superior facet-midpedicle axis distance, 9.9 to 12.0 mm for inferior facet-midpedicle axis distance, 42.3 degrees to 51.5 degrees for transverse angle, and 5.2 degrees to 14.1 degrees for sagittal angle. CONCLUSIONS: Linear measurements of pedicle dimensions and also axial angles from horizontal and vertical planes may provide some anatomic limitations for subaxial cervical transpedicular screw fixation, and also contribute to the safety of the surgical procedure. One should also rely on tomographic data and computer assisted guidance systems. PMID- 15534409 TI - Case report: whiplash-associated disorder from a low-velocity bumper car collision: history, evaluation, and surgery. AB - STUDY DESIGN: Case report of a patient with a whiplash-associated disorder following a bumper car collision. Imaging studies failed to provide an anatomic explanation for the debilitating symptoms. OBJECTIVES: To report a chronic, debilitating pain syndrome after a low-velocity bumper car collision while using complex range-of-motion data for the diagnosis, prognosis, and surgical indication in whiplash-associated disorder. SUMMARY OF BACKGROUND DATA: The controversy of whiplash-associated disorder mainly concerns pathophysiology and collision dynamics. Although many investigations attempt to define a universal lesion or determine a threshold of force that may cause permanent injury, no consensus has been reached. METHODS: Eight years after a low-velocity collision, the patient underwent surgical excision of multiple painful trigger points in the posterior neck. Computerized motion analysis was used for pre- and postoperative evaluations. RESULTS: Surgical treatment resulted in an increase in total active range of motion by 20%, reduced intake of pain medication, doubled the number of work hours, and generally led to a dramatic improvement in quality of life. CONCLUSIONS: This case of whiplash-associated disorder after a low-velocity collision highlights the difficulty in defining threshold of injury in regard to velocity. It also illustrates the value of computerized motion analysis in confirming the diagnosis of whiplash-associated disorder and in the evaluation of prognosis and treatment. PMID- 15534410 TI - A pilot study evaluating the safety and efficacy of OP-1 Putty (rhBMP-7) as a replacement for iliac crest autograft in posterolateral lumbar arthrodesis for degenerative spondylolisthesis. AB - STUDY DESIGN: A prospective, randomized, controlled, multicenter clinical study was conducted. OBJECTIVE: To compare the clinical and radiographic outcomes of patients treated with OP-1 (BMP-7) Putty to autogenous iliac crest bone graft for one-level uninstrumented posterolateral fusion of the lumbar spine following decompressive laminectomy for the treatment of symptomatic degenerative spondylolisthesis with spinal stenosis. BACKGROUND: Preclinical studies have demonstrated that osteoinductive recombinant human osteogenic protein 1 in the form of OP-1 Putty is successful at achieving a posterolateral fusion in rabbits and dogs without any significant safety concerns. METHODS: Thirty-six patients with degenerative lumbar spondylolisthesis and symptoms of neurogenic claudication were randomized (2:1) to either OP-1 Putty (3.5 mg of OP-1 per side) or autogenous iliac crest bone graft for one-level uninstrumented posterolateral fusion following a decompressive laminectomy. Enrollment in the study was complete when 24 OP-1 Putty patients and 12 autograft patients had been randomized and treated. A patient administered Oswestry scale and SF-36 scale were used to determine clinical outcomes. Independent, blinded neuroradiologists reviewed both static and dynamic radiographs to determine fusion status. RESULTS: At the 1-year follow-up, 32 patients were available for clinical analysis and 29 patients were available for radiographic review. Clinical success as measured on the Oswestry scale was achieved by 18 of 21 (86%) OP-1 Putty patients and 8 of 11 (73%) autograft patients. SF-36 pain index scores showed similar results. Fourteen of 19 (74%) OP-1 Putty patients and 6 of 10 (60%) autograft patients achieved a successful posterolateral fusion fulfilling all fusion criteria. Of the 29 evaluable patients, 15 were both clinical and radiographic successes, 5 were radiographic successes but were clinical failures, 1 patient was both a radiographic and clinical failure, and 8 patients were radiographic failures but were clinical successes. No systemic toxicity, ectopic bone formation, recurrent stenosis, or other adverse events related to the OP-1 Putty implant were observed. CONCLUSION: Although the posterolateral spine is a challenging fusion environment in patients with degenerative spondylolisthesis, successful radiographic fusion was obtained using OP-1 Putty at a rate that was similar to autograft given the number of patients in this study. Importantly, there were no apparent adverse consequences related to the use of the OP-1 Putty implant in this patient population. PMID- 15534411 TI - Good outcome and restoration of lordosis after anterior lumbar interbody fusion with additional posterior fixation. AB - STUDY DESIGN: Fifty-two patients with degenerative disc disease underwent single- or double-level anterior lumbar interbody fusion with SynCage and additional posterior fixation as treatment for degenerative disc disease and were prospectively followed for 4 years. OBJECTIVES: To test the clinical performance of anterior lumbar interbody fusion with SynCage, with emphasis on the safety and efficacy of the surgical procedure and the ability to restore anatomy and fuse the motion segment. SUMMARY OF BACKGROUND DATA: Anterior lumbar interbody fusion using femoral allograft and/or autologous bone has a high complication rate. With cage technology, some of these complications can be avoided. The design characteristics of the SynCage offer advantages in restoring and maintaining intervertebral height and restoration of lumbar lordosis. METHODS: Thirty-three patients underwent single-level and 19 patients double-level anterior lumbar interbody fusion with SynCage and additional posterior fixation (translaminar screws, n = 32 or pedicle screws, n = 10). Radiologic and functional results (VAS and Oswestry score) were evaluated. RESULTS: Intervertebral height was corrected from an average of 8.7 to 17.6 mm. Lordosis of the fused segment was significantly increased (average 10.6 degrees for the fused segment and 8 degrees for lumbosacral lordosis). After 2 years, there was radiologic evidence for fusion in 70 of 71 (98.6%) levels. Functional scores showed a significant improvement in pain and function up to the 2-year follow-up observation. At the 4 year follow-up observation, there was some loss of the initial improvement in VAS and Oswestry scores. Despite this loss, they remained significantly better than the preoperative scores. CONCLUSIONS: Anterior lumbar interbody fusion with SynCage and additional posterior fixation is a safe and effective procedure. Intervertebral height is corrected, and lumbosacral lordosis is restored. An initial improvement in VAS and Oswestry scores is partly lost at the 4-year follow-up observation, but 4-year results are still significantly better than the preoperative scores. PMID- 15534413 TI - The management of acute thoracolumbar burst fractures with anterior corpectomy and Z-plate fixation. AB - STUDY DESIGN: A retrospective review of a consecutive series of patients with acute thoracolumbar burst fractures who were surgically treated with an anterior corpectomy and fusion with anterolateral Z-plate fixation. OBJECTIVES: To evaluate the clinical and radiographic success of the management of acute thoracolumbar burst fractures by corpectomy, structural grafting, and anterolateral internal fixation. SUMMARY OF BACKGROUND DATA: Burst fractures are frequently associated with instability or neurologic deficit. Modern surgical procedures for these fractures have been performed via both anterior and posterior approaches. Anterior surgical treatment allows direct decompression of the neural elements and correction of deformity. Newer anterior instrumentation devices, combined with a structural graft, allow a stable construct that may obviate a posterior procedure. An anterior procedure generally requires fusion of only two levels compared to posterior fusion, which generally requires more. METHODS: A retrospective review of a consecutive series of patients with thoracolumbar burst fractures treated with anterior surgery, strut graft, and fixation with a Z-plate was carried out. Fractures were considered acute if surgically treated within 30 days. Clinical and radiographic evaluation was performed on all 35 patients with acute thoracolumbar burst fractures. Surgical indications were incomplete neurologic deficit, segmental kyphotic deformity, or significant comminution. All patients with acute thoracolumbar burst fractures with spinal cord injury were treated with an intravenous steroid protocol and were operated on within 24 hours of admission unless medically precluded. Forty six percent (16 of 35) of patients with acute thoracolumbar burst fractures presented with a neurologic deficit. RESULTS: All 16 patients with neurologic deficit demonstrated at least one Frankel grade improvement on final observation, with 11 (69%) patients demonstrating complete neurologic recovery. Thirty-three patients were treated with anterolateral instrumentation only. Twenty-nine of thirty patients demonstrated radiographic healing. Five were lost to follow-up observation. One patient required subsequent posterior fusion for increasing kyphotic deformity. There were no instances of hardware failure. Sagittal alignment was improved from a mean preoperative kyphosis of 18 degrees to 6 degrees at final follow-up observation. CONCLUSIONS: Anterior corpectomy, strut graft, and Z-plate fixation is an effective treatment for thoracolumbar burst fractures. It allows direct decompression of the spinal cord in the acute setting and was associated with a high rate of neurologic improvement, no instances of neurologic worsening in any case, and a low complication rate. PMID- 15534415 TI - A new clinical evaluation for hysterical paralysis. AB - STUDY DESIGN: A cross-sectional study was performed to elucidate the usefulness of a new clinical evaluation, and a prospective study was performed to detect hysterical paralysis using this evaluation method. OBJECTIVES: To make a correct diagnosis of hysterical paralysis, a new clinical evaluation was developed. SUMMARY OF BACKGROUND DATA: Hysterical paralysis is a conversion disorder. Its diagnosis must be ruled out when encountering a patient with paralysis, therefore imaging and electrophysiological studies are often necessary, but costly. The principal salient diagnostic features for diagnosing hysterical paralysis are thought to be the preservation of a normal reflex pattern, normal rectal sensation, and normal bladder and bowel functions; however, these features are not always successfully identified. METHODS: A new clinical evaluation named the "Spinal Injuries Center" test was developed. The lower extremities of the patients were divided into two groups as follows: in group A, the patients were able to lift up the knee; in group B, the patients were unable to lift up the knee. The 96 legs of the 48 patients who had obvious myelomalacia were randomly chosen. All legs were investigated using the Spinal Injuries Center test, and the association between each group and the Spinal Injuries Center test was examined. The 28 legs of the 14 patients in whom hysterical paralysis was diagnosed were prospectively evaluated using the Spinal Injuries Center test, and the association between the groups and the Spinal Injuries Center test was examined. RESULTS: Forty-eight legs were classified as group A, and 48 legs were classified as group B. In group A, 45 legs were judged to be positive for the Spinal Injuries Center test, and 3 legs were negative. In group B, 1 leg was judged to be positive for the Spinal Injuries Center test, and 47 legs were negative. All legs of the patients with hysterical paralysis were classified as group B; however, all legs were positive for the Spinal Injuries Center test. CONCLUSIONS: The Spinal Injuries Center test is a new clinical evaluation method that can help make a correct diagnosis of hysterical paralysis. When a patient is unable to lift up his knees by himself, the result of the Spinal Injuries Center test is considered to be positive, and hysterical paralysis is diagnosed in such patients. The diagnosis of hysterical paralysis must be ruled out when encountering patients with paralysis, and as a result, imaging and electrophysiological studies are often necessary. Unfortunately, such tests are costly. Thus, a new clinical evaluation for the diagnosis of hysterical paralysis, named the Spinal Injuries Center test, was developed. When patients who are unable to lift up their knees by themselves test positive using the Spinal Injuries Center test, then they are considered to have ether hysterical or simulated paralysis. PMID- 15534417 TI - A possible methodological flaw in comparing dominant and nondominant sided lumbar spine muscle responses without simultaneously considering hand dominance. AB - STUDY DESIGN: Different factorial designs analyzing the same data were compared for consistent interpretation. OBJECTIVE: To assess hand dominance as an explanatory factor when comparing dominant and nondominant side back muscle response time delay and fatigability. SUMMARY OF BACKGROUND DATA: A number of studies have evaluated lumbar spinal muscle responses to sudden load and fatigue. Although the dominant side of the back is usually determined by considering hand dominance, few studies explicitly consider hand dominance as part of the explanatory model and assume that right- and left-handed patients are not unique cohorts. METHODS.: The erector spinae and multifidi lumbar muscle groups in patients with chronic low back pain (CLBP) were investigated. For the response time to a sudden load, the biceps brachii muscle was compared with erector spinae and multifidi muscles to understand any compensatory upper extremity movement patterns. For fatigue, the erector spinae and multifidi muscle median frequency (MF) and its slope (MFS) were measured during a 1-minute isometric back extension maneuver. RESULTS: Altogether, 46 patients with CLBP were studied. Right- and left-handed subjects did not differ in reported disability based on the Oswestry Disability Index, (F1, 44 = 2.11, P < 0.153). The dominant and nondominant side response times were significantly different for left-handed patients but not for right-handed patients. The nondominant side back muscles, located on the contralateral side of the dominant hand, were faster for the right-hand dominant patients and slower for the left-hand dominant patients. In the muscles on the nondominant side of the back, the left-hand dominant patients demonstrated significantly more fatigue than right-hand dominant patients. However, in the muscles on the dominant side of the back, neither set of patients demonstrated fatigability. CONCLUSIONS: When examining differential lumbar muscle responses, failing to consider hand dominance explicitly in conjunction with the side assessed resulted in confounding effects. The results of the study indicated a delayed back muscle response time on the nondominant side. This delayed response time raises the possibility that unbalanced muscle activity could prompt a decreased, uncoordinated, asymmetric bracing effect, thereby increasing the risk of lumbar segment buckling. Further studies are needed to investigate the characteristics of the back muscles on the same side of the dominant hand and the factors mediating neuromuscular differences in patients with CLBP. Understanding the effects of hand dominance on fatigue in certain back muscles should lead to better treatments thereby improving neuromuscular control in patients with CLBP. PMID- 15534418 TI - The core outcomes for neck pain: validation of a new outcome measure. AB - STUDY DESIGN: Patients enrolled in a randomized controlled trial were asked to complete various questionnaires, which were then compared to establish validity for a new neck pain questionnaire. OBJECTIVES: To validate a new and brief outcome measure for use with patients with mechanical neck pain. SUMMARY OF BACKGROUND DATA: Neck pain is a very common problem and one that clinicians will be required to treat with some regularity. In today's climate of evidence-based practice and the need to quantify and justify clinical intervention, a quick and easy method to evaluate progress is required. Such a measure has already been produced for those experiencing back pain, but as yet, there is no such measure for neck pain and this needs to be addressed. METHODS: The back pain measure was adapted to enable its use with patients with neck pain. Repeatability was assessed by using a 1-week test/retest on 104 patients who were enrolled in a neck pain trial. Validity was assessed by comparing the new questionnaire against other already well validated measures (i.e., the Neck Disability Index and a Visual Analogue Scale for pain) with 133 patients. RESULTS: The test/retest showed excellent repeatability with high intraclass correlations and P < 0.001 for each question tested. The Core Neck Pain Questionnaire also showed good validity, giving close agreement to the other comparison measures. CONCLUSION: The short Core Neck Pain Questionnaire has been demonstrated to be repeatable and valid as a brief outcome measure for use with patients with mechanical neck pain. PMID- 15534419 TI - The effect of previous low back surgery on general health status: results from the National Spine Network initial visit survey of patients with low back pain. AB - STUDY DESIGN: A cross-sectional study on 18,325 patients with back pain enrolled at first visit in the National Spine Network (NSN) database from January 1998 to April 2000. OBJECTIVES: To examine whether patients who had previous low back surgeries had poorer general health status than patients with no surgery. SUMMARY OF BACKGROUND DATA: Several studies have described the role of psychological abnormalities in patients with chronic low back pain. Some of these patients have had previous spinal surgeries performed. No study has examined the effects of previous low back surgery on the general health status. METHODS: The Short Form Health Survey 36 was administered to the initial visit NSN patients. Of the 18,325 patients enrolled, 3,632 had previous low back surgeries. RESULTS: Patients who had previous lumbar surgeries fared significantly poorly in all 10 scores of the SF-36 health survey, even after adjustment for confounding factors. Among these patients, decompression achieved significantly higher scores for General Health, Role-Physical, and Mental Component Summary scales. Patients who had decompression as their most recent surgery had higher scores for General Health, Role-Physical, Role-Emotional, and Mental Component Summary scales, when compared to those who had other surgeries. Patients who had instrumentation as their most recent surgery had higher scores for Bodily Pain and Physical Component Summary scores. There is a positive correlation between time since last surgery and the SF-36 outcomes. CONCLUSIONS: Previous back surgery is associated with significantly worse general health status than those without surgery. Among patients who had previous surgeries, decompression seems to exert better effects on SF-36 health status. There is a positive correlation between time since last surgery and the SF-36 outcomes, although the SF-36 scores are significantly lower than those without previous surgery. PMID- 15534420 TI - Adjacent segment disease after lumbar or lumbosacral fusion: review of the literature. AB - STUDY DESIGN: Review of the literature. OBJECTIVES: Review the definition, etiology, incidence, and risk factors associated with as well as potential treatment options. SUMMARY OF BACKGROUND DATA: The development of pathology at the mobile segment next to a lumbar or lumbosacral spinal fusion has been termed adjacent segment disease. Initially reported to occur rarely, it is now considered a potential late complication of spinal fusion that can necessitate further surgical intervention and adversely affect outcomes. METHODS: MEDLINE literature search. RESULTS: The most common abnormal finding at the adjacent segment is disc degeneration. Biomechanical changes consisting of increased intradiscal pressure, increased facet loading, and increased mobility occur after fusion and have been implicated in causing adjacent segment disease. Progressive spinal degeneration with age is also thought to be a major contributor. From a radiographic standpoint, reported incidence during average postoperative follow up observation ranging from 36 to 369 months varies substantially from 5.2 to 100%. Incidence of symptomatic adjacent segment disease is lower, however, ranging from 5.2 to 18.5% during 44.8 to 164 months of follow-up observation. The rate of symptomatic adjacent segment disease is higher in patients with transpedicular instrumentation (12.2-18.5%) compared with patients fused with other forms of instrumentation or with no instrumentation (5.2-5.6%). Potential risk factors include instrumentation, fusion length, sagittal malalignment, facet injury, age, and pre-existing degenerative changes. CONCLUSION: Biomechanical alterations likely play a primary role in causing adjacent segment disease. Radiographically apparent, asymptomatic adjacent segment disease is common but does not correlate with functional outcomes. Potentially modifiable risk factors for the development of adjacent segment disease include fusion without instrumentation, protecting the facet joint of the adjacent segment during placement of pedicle screws,fusion length, and sagittal balance. Surgical management, when indicated, consists of decompression of neural elements and extension of fusion. Outcomes after surgery, however, are modest. PMID- 15534421 TI - Early experiences with video-assisted thoracoscopic surgery: our first 70 cases. AB - STUDY DESIGN: Prospective consecutive series. OBJECTIVE: Analysis of the results and outcomes of patients treated with video-assisted thoracoscopic surgery for spinal pathology. SUMMARY OF BACKGROUND DATA: Video-assisted thoracoscopic surgery is an alternative to open thoracotomy. It has been suggested that the learning curve is substantial. The authors present their early experience in treating a variety of spinal pathologies with this technique. METHODS: Seventy cases were available at the 2-year follow-up. Video-assisted thoracoscopic surgery with the goal of anterior spinal release and fusion was carried out on patients with the following diagnoses: idiopathic scoliosis, neuromuscular spinal deformity, Scheuermann kyphosis, congenital and infantile scoliosis, neurofibromatosis, Marfan syndrome, postradiation scoliosis, and repair of pseudarthrosis. Three patients had excision of the first rib to treat thoracic outlet syndrome. Two patients had excision of intrathoracic neurofibroma and a benign rib tumor. One had anterior fusion following thoracic spine fracture dislocation. RESULTS: The average operative time for the thoracoscopic anterior release with discectomy and fusion procedure was 256 minutes (range 150-405 minutes). The average number of discs excised was 8 (range 4-11 discs). The average operative time per disc was 32.5 minutes (range 20-45 minutes). The average blood loss during the thoracoscopic anterior release with diskectomy and fusion was 285 mL (range 150-405 mL). Final postoperative scoliosis and kyphosis corrections were 68% (range 41-91%) and 90% (range 47-100%), respectively. Complications related to thoracoscopy occurred in 3 patients. All deformity patients had evidence of anterior fusion radiographically. CONCLUSION: Video assisted thoracoscopic surgery provides a safe and effective alternative to open thoracotomy in the treatment of thoracic pediatric spinal deformities. The procedure remains time consuming. PMID- 15534423 TI - Historical perspective: William Adams, the forward bending test, and the spine of Gideon Algernon Mantell. AB - William Adams described the Forward Bending Test for scoliosis in 1865. His understanding of the nature of the rotational element of scoliosis was given by a postmortem he performed on an eminent surgeon and geologist, Gideon Mantell. The clinical history of Dr. Mantell is well documented. PMID- 15534425 TI - Focus on the fire of life. PMID- 15534426 TI - Variability in energy expenditure and its components. AB - PURPOSE OF REVIEW: To review factors contributing to variation in total daily energy expenditure and its primary components: (1) resting metabolic rate; (2) diet-induced thermogenesis; and (3) activity thermogenesis, including exercise energy expenditure and nonexercise activity. For each component, the expected magnitude of intra-individual variability is also considered. We also reviewed studies that quantified the variability in 24 h energy expenditure. RECENT FINDINGS: In humans, the coefficient of variation in the components of total daily energy expenditure is around 5-8% for resting metabolic rate, 1-2% for exercise energy expenditure, and around 20% for diet-induced thermogenesis. The coefficient of variance for 24 h energy expenditure measured using a room calorimeter for resting metabolic rate is around 5-10%. Thus, these measures are all rather reproducible. Total daily energy expenditure varies several-fold in humans, not due to variation in resting metabolic rate, diet-induced thermogenesis, or exercise thermogenesis, but rather, due to variations in nonexercise activity. A variety of factors impact nonexercise activity, including occupation, environment, education, genetics, age, gender, and body composition, but little is known about the magnitude of effect. SUMMARY: Resting metabolic rate, diet-induced thermogenesis, exercise energy expenditure, and 24 h energy expenditure are highly reproducible. Coefficient of variation is smallest for exercise energy expenditure, followed by resting metabolic rate, 24 h energy expenditure, and diet-induced thermogenesis. There is considerable variability in total daily energy expenditure, largely due to variations in nonexercise activity. Although the factors that impact upon nonexercise activity are understood, their contribution to variation in total daily energy expenditure is unclear. PMID- 15534427 TI - Physical activity and human energy expenditure. AB - PURPOSE OF REVIEW: This is a review on the measurement of physical activity under daily life conditions. The focus is on the doubly labelled water method and accelerometry. Doubly labelled water is a gold standard and the reference for the validation of field methods to assess physical activity. Accelerometry is the most objective and precise technique to assess activity patterns in terms of frequency, duration and intensity. Applications of the two techniques are illustrated with the limits of physical activity and energy expenditure and with activity intensity as a determinant of the physical activity level. RECENT FINDINGS: The upper limit of the physical activity index (total energy expenditure as a multiple of basal metabolic rate) of 2.5, as derived from cross sectional data, is confirmed by training intervention studies. Exercise training, in which total energy expenditure was measured before and at the end of the training programme, showed no increase in physical activity index when training was combined with an energy restricted diet and in elderly subjects. In children, the distribution of time spent at activities with low and high intensity determines the physical activity index while in adults moderate-intensity activities are the main determinant. SUMMARY: In adults, within the normal physical activity index range, the distribution of time spent at activities with low and moderate intensity determines the physical activity level. High-intensity activity does not have much impact on daily energy expenditure. High-intensity activity is not required to increase the activity energy expenditure. PMID- 15534428 TI - Energy restriction and aging. AB - PURPOSE OF REVIEW: The focus of this review is on current research involving long term calorie restriction and the resulting changes observed in possible biomarkers of aging. Special emphasis will be given to the basic and clinical science studies which are currently investigating the effects of controlled, high quality energy-restricted diets on both biomarkers of longevity and on the development of chronic diseases related to age and obesity in humans. RECENT FINDINGS: Prolonged calorie restriction has been shown to extend both the median and maximal lifespan in a variety of lower species such as yeast, worms, fish, rats, and mice. Mechanisms of this lifespan extension via calorie restriction are not fully elucidated, but possibly involve significant alterations in energy metabolism, oxidative damage, insulin sensitivity, and functional changes in both the neuroendocrine and sympathetic nervous systems. Ongoing studies of prolonged energy restriction in humans are now making it possible to analyze changes in these aging biomarkers to unravel some of the mechanisms of its antiaging phenomenon. SUMMARY: With the incremental expansion of research endeavors in the area of energy or calorie restriction, data on the effects of calorie restriction in animal models and humans are becoming more accessible. Detailed analyses from controlled human trials involving long-term calorie restriction will allow investigators to link observed alterations in body composition down to changes in molecular pathways and gene expression, with their possible effects on the biomarkers of aging. PMID- 15534429 TI - Metabolic consequences of overfeeding in humans. AB - PURPOSE OF REVIEW: Overfeeding leads to obesity and metabolic disorders, including impaired glucose homeostasis, lipid disorders, and hepatic steatosis. The consequences of standardized overfeeding on body weight have shown, however, considerable interindividual variability, which suggests that it also leads to adaptative changes in energy expenditure, in some individuals at least. The present review is mainly focused on the recent developments regarding the effects of overfeeding on energy expenditure. RECENT FINDINGS: Individuals who gain the less body weight during overfeeding are those who experience a greater increase in total energy expenditure. This increase in energy expenditure has been attributed to stimulation of nonexercise physical activity. Recent developments regarding adaptative increases in physical activity are critically reviewed. Overfeeding also alters the pathways used for carbohydrate storage after a glucose load, by increasing de-novo lipogenesis in the liver and adipose tissue at the expense of glycogen storage. The sympathetic nervous system is a good candidate for energy expenditure increase during overfeeding. The increases in energy expenditure observed during acute stimulation of the sympathetic nervous system were however found to be unaltered by short-term overfeeding. SUMMARY: The mechanisms by which some individuals protect themselves against body weight gain remain poorly understood. Nonvoluntary physical activity may allow one to increase energy expenditure during overfeeding, and may therefore constitute a regulatory factor in body weight control. The biological determinant of spontaneous, nonvoluntary physical activity, however, remains to be investigated. PMID- 15534430 TI - Leptin and energy expenditure. AB - PURPOSE OF REVIEW: A fundamental advance in our understanding of endocrine control of energy balance and body weight came with the discovery of the adipocyte-derived hormone leptin. The leptin pathway appeared to be the long sought peripheral signal pathway from the adipose tissue to the brain involved in the regulation of feeding and energy balance. RECENT FINDINGS: Initially, leptin was considered to function as the long-sought antiobesity hormone. According to this hypothesis, rising concentrations of leptin with increasing adiposity would generate a signal to reduce food intake and increase energy expenditure in order to limit further weight gain. However, widespread resistance to the proposed antiobesity action of leptin is observed in humans, which might reflect the fact that the inability to store energy efficiently at times of abundance is evolutionarily disadvantageous. According to this alternative view, falling leptin concentrations observed during fasting act as a peripheral signal of starvation, which serves to conserve energy in the face of limited reserves. However, leptin administration failed to blunt the changes in energy expenditure during severe energy restrictions in several clinical studies. In addition, leptin therapy in several different human low-leptin states failed to affect energy expenditure in recent studies. SUMMARY: Increasing evidence from human studies suggests that leptin predominantly influences the human energy balance through appetite but appears not to be involved in regulating energy expenditure. None of the expected factors such as resting metabolic rate, total diurnal energy expenditure or dietary induced thermogenesis was related to blood leptin concentrations. PMID- 15534431 TI - Energy expenditure in chronic spinal cord injury. AB - PURPOSE OF REVIEW: Obesity is a common secondary complication of chronic spinal cord injury and is associated with adverse metabolic sequelae. Because positive energy balance is the fundamental cause of obesity, we herein review the current knowledge pertaining to total daily energy expenditure, including resting metabolic rate, the thermic effect of food, and physical activity, in the spinal cord injury population. RECENT FINDINGS: Commonly used equations to predict resting metabolic rate overestimate measured requirements in chronic spinal cord injury by 5-32%. Measured resting metabolic rate is 14-27% lower in persons with spinal cord injury versus those without, due to decreased fat-free mass and sympathetic nervous system activity in this population. However, preliminary evidence suggests that neither the metabolic activity of the fat-free body, nor the obligatory phase of the thermic effect of food is different between those with and without injury. Physical activity levels, especially in those with tetraplegia and complete lesions, are lower than recommended or lower than those of able-bodied persons. SUMMARY: New equations to predict resting metabolic rate should be validated and prospectively tested in a large sample of men and women with complete and incomplete paraplegia and tetraplegia. Whether the facultative phase of the thermic effect of food is different between those with and without SCI remains to be elucidated. Persons with chronic spinal cord injury, and perhaps those with tetraplegia and complete lesions especially, should be encouraged to engage in increased frequency, intensity and/or duration of physical activity. Future research efforts should explore the effects of level and completeness of neurological lesion on resting metabolic rate, thermic effect of food, and physical activity. PMID- 15534432 TI - Physical activity: the health benefits outweigh the risks. AB - PURPOSE OF REVIEW: This article will summarize the current findings on the effects of physical activity on human health and well-being. RECENT FINDINGS: Physical activity is associated with enhanced health and reduced risk of all cause mortality such as cardiovascular disease, hypertension, type 2 diabetes, obesity, osteoporosis, sarcopenia, cognitive disorders, and some forms of cancer. Nevertheless, the effects of exercise with respect to potential health consequences are complex. When untrained or previously sedentary persons undertake vigorous exertion suddenly, the undesired side effects of injuries, dehydration or cardiac arrest are amplified. SUMMARY: It is reasonable to conclude that the risk exposure through physical activity is outweighed by its overall benefits, and health authorities strongly encourage participation in moderate intensity physical activity on a daily basis. In the future, the identification and characterization of particularly inactive sub-groups of the population may facilitate and optimize the planning of public health interventions. PMID- 15534433 TI - Skeletal benefits of soy isoflavones: a review of the clinical trial and epidemiologic data. AB - PURPOSE OF REVIEW: Osteoporosis is a worldwide problem of immense magnitude that is expected to worsen in many countries with aging populations. Consequently, there is a need to identify ways to reduce the risk of developing this disease. This is especially true in light of clinical trial data showing the long-term harm of conventional hormone therapy outweighs the benefits. It is well established that many dietary components impact the skeletal system; in this regard there is particular interest in the possible skeletal benefits of soybean isoflavones. The purpose of this review is to evaluate the clinical and epidemiologic studies relevant to the hypothesis that isoflavones promote bone health. RESULTS: Fifteen clinical trials were identified that examined the effects of isoflavones or isoflavone-rich soy protein on bone mineral density. Most trials were conducted for 1 year or less and involved relatively few (<30) participants per group. The findings from these studies are inconsistent but generally suggest that isoflavones reduce bone loss in younger postmenopausal women. Similarly, the limited epidemiologic data generally show that among Asian populations isoflavone intake is associated with higher bone mineral density. The clinical data suggest that approximately 80 mg/day isoflavones are needed to derive skeletal benefits whereas the epidemiologic data suggest lower amounts are efficacious. SUMMARY: Until more definite data are available, although soy foods and isoflavones can not be viewed as substitutes for established anti osteoporotic medications health professionals can feel justified in encouraging postmenopausal women concerned about bone health to incorporate soyfoods into their diet. PMID- 15534434 TI - Folate and brain function in the elderly. AB - PURPOSE OF REVIEW: Over the past several decades, folate has emerged as an important nutrient in several key conditions of concern to the elderly. Subclinical levels of folate inadequacy can have significant negative impacts on health in older individuals. RECENT FINDINGS: Serum and red blood cell folate levels are associated with depression in younger individuals, but the relationship is less clear in older people. However, folate status does predict response to antidepressant treatment in older individuals. Cognitive decline and some forms of dementia, including Alzheimer's disease, are associated with lower folate levels. Supplementation with folic acid can provide cognitive benefits in some circumstances. Folic acid supplementation is generally regarded as safe; however, there remains some concern that high levels of folic acid may exacerbate the neurological consequences of a vitamin B12 deficiency. SUMMARY: Evidence for the role of folate in depression and dementia in the aged is increasing, although there remains much about mechanisms to be determined. PMID- 15534436 TI - Bibliography. Current world literature. Nutrition and physiological function. PMID- 15534435 TI - Intake of fruit and vegetables and risk of stroke: an overview. AB - PURPOSE OF REVIEW: This review presents recent scientific advances in understanding the role of fruit and vegetables in lowering stroke risk, based on epidemiological and clinical studies. RECENT FINDINGS: Recent studies indicate that high dietary intake of fruit and vegetables is associated with reduced risk of stroke and other cardiovascular outcomes. Both observational studies and interventional trials have consistently shown this inverse relationship. Multiple mechanisms may underlie the apparent beneficial effect. SUMMARY: Public health policy should promote increased dietary intake of fruit and vegetables, as this may have a substantial positive effect on stroke prevention. PMID- 15534438 TI - Recent advances in the management of placenta previa. AB - PURPOSE OF REVIEW: Despite the widespread and routine use of ultrasound to make the diagnosis of placenta previa, evidence-based classification and management strategies have failed to evolve over the years. The purpose of this review is to present the current evidence supporting the screening, diagnosis and management of placenta previa. RECENT FINDINGS: The prevalence of placenta previa is significantly overestimated due to the practice of routine mid-pregnancy scan, and many women currently undergo a repeat scan in late pregnancy for placental localization. Recent reports support limiting third-trimester scans to only those cases where the placental edge either reaches or overlaps the internal cervical os at 20-23 weeks of pregnancy. In some cases of mid-trimester placenta previa, the placental edge is more likely to "migrate" than others, and it appears that ultrasound may be useful to predict this process. At term, women with placental edge within 2 cm of the internal cervical os require a Caesarean section for delivery, whereas an attempt at vaginal birth is appropriate if this distance is more that 2 cm. Ultrasound also has a role in the diagnosis and management of both vasa previa and placenta accreta. SUMMARY: This review addresses screening for placenta previa. A simple and pragmatic ultrasound classification of placenta previa and low-lying placenta is proposed. Caesarean section is recommended for delivery in cases of placenta previa. Women with a low-lying placenta have at least 60% chance of a vaginal birth, but should be monitored for post-partum haemorrhage. Vasa previa is a rare complication but antenatal diagnosis is possible. It should particularly be suspected in in-vitro fertilization conceptions, and where the placental edge covers the os in mid-pregnancy but recedes later on. Prenatal diagnosis of placenta accreta should be based on the placental lacunae signs rather than the absence of retro-placental clear space. PMID- 15534439 TI - Thrombophilia and adverse pregnancy outcome. AB - PURPOSE OF REVIEW: Recent case-control studies and metaanalyses have attempted to quantify the risks associated with individual thrombophilic defects and adverse clinical events in pregnancy, including fetal loss, preeclampsia, placental abruption and intrauterine growth restriction. This review has examined the evidence. RECENT FINDINGS: The literature is in general agreement that thrombophilia increases the risk of venous thromboembolism and adverse pregnancy outcomes, including pregnancy loss, preeclampsia, placental abruption and intrauterine growth restriction in pregnancy. However, the size of the estimated risks varies between individual studies due to heterogeneity in study design. Low molecular-weight heparin has been shown to be the superior choice, on the grounds of safety and effectiveness, in preventing venous thromboembolism and improving pregnancy loss. Large-scale, randomized controlled studies are required, however, to confirm these findings. Although selective thrombophilia screening based on prior venous thromboembolism history has been shown to be marginally more cost effective than universal screening in pregnancy, the overall clinical and economic benefit of universal and selective screening is unsupported. SUMMARY: Despite the growing evidence in the literature, there are still gaps in our knowledge of thrombophilia and pregnancy. In particular, accurate estimates are required of the risks of venous thromboembolism and adverse pregnancy outcomes associated with some thrombophilias and the relative clinical and cost effectiveness of different anticoagulation therapies in the prevention of venous thromboembolism and pregnancy loss. More large-scale studies are required to better inform clinicians and help determine optimum management and prevention strategies of thrombophilia and associated adverse clinical events in pregnancy. PMID- 15534440 TI - Tocolysis and preterm labour. AB - PURPOSE OF REVIEW: There is persisting controversy about tocolytic treatment for preterm labour. This review addresses this controversy by appraising the recent clinical literature. RECENT FINDINGS: Surveys of obstetricians indicate a high usage of tocolysis for preterm labour, but evidence that this treatment confers overall benefit is still lacking. Betamimetics are now, correctly, being abandoned in favour of nifedipine, which has superior tocolytic properties and better neonatal outcomes. There is no evidence of effectiveness for magnesium sulphate as a tocolytic. Atosiban is a newer agent, which appears to be effective in delaying preterm birth with a favourable maternal safety profile, but there are persisting concerns about the lack of impact on perinatal mortality and morbidity. Current research is addressing the COX-2 inhibitor, rofecoxib, which has theoretical advantages with respect to fetal safety. SUMMARY: For the relatively small proportion of women in otherwise uncomplicated preterm labour prior to 34 weeks' gestation, there appears to be a place for short-term tocolysis to gain time so that corticosteroids can be administered to enhance fetal lung maturation and, if necessary, to transfer the woman to a facility with a neonatal intensive care unit. Nifedipine is an effective and cheap tocolytic agent. Atosiban appears to also be effective, but it is expensive and not universally available. Betamimetics and magnesium sulphate should be abandoned as tocolytic agents. There is a need for further clinical trials to establish an unequivocal evidence base for tocolysis, which requires placebo-controlled trials, and for comparative trials to identify the agent with superior characteristics. PMID- 15534441 TI - Sperm washing, use of HAART and role of elective Caesarean section. AB - PURPOSE OF REVIEW: Today, 50% of people living with HIV are women and most have been sexually infected. Highly active antiretroviral therapy (HAART) reduces the rates of both sexual and vertical infection, but maximum protection is achieved with sperm washing and elective Caesarean section. RECENT FINDINGS: Men taking HAART have lower seminal concentration of HIV, and sexual transmission may be reduced. However, a certain percentage of aviraemic men retain viral presence in semen, and unprotected intercourse to achieve fertilization must be discouraged as it carries the risk of sexual transmission of the virus. HIV-discordant couples should be informed that sperm washing can remove HIV from semen, allowing conception without the risk of infection for the seronegative female and eventually the child. In HIV-positive women, perinatal transmission of HIV can be curtailed to less than 2% by using HAART to decrease maternal viral load and offering prenatal preexposure prophylaxis of the fetus, and elective Caesarean section. Each intervention carries specific risks and benefits. The contribution of each preventive arm in achieving fetal protection can only be crudely measured and optimal obstetric management must involve discussion with the pregnant woman of the pros and cons of each strategy. SUMMARY: In HIV-positive men taking HAART, seminal viral load is decreased but not eliminated and fertilization should be achieved through sperm washing to offer maximum protection for the uninfected female. Pregnant HIV-positive women on antiretroviral medication have a reduced risk of transmitting the virus, but should still be counselled about the possibility to further limit the chances of infecting their infant through elective Caesarean section. PMID- 15534442 TI - HIV and pregnancy: is the outlook for mother and baby transformed? AB - PURPOSE OF REVIEW: Advances in antiretroviral regimens and specific obstetrical procedures have enabled HIV-positive women to have children, with a very low risk of transmitting the infection to the infant and with improved chances of seeing their children reach adulthood. New studies have given providers of care better information on how to assist women with HIV who want to have a child in the safest possible way. RECENT FINDINGS: Highly active antiretroviral therapy can effectively control viral replication and reduce the risk of vertical transmission. The benefit of treatment for the mother and the infant must be balanced against any negative effects on pregnancy, the embryo and the fetus. Potential long-term consequences of prenatal exposure to potent compounds should also be considered and monitored. The evidence suggests that even in women with undetectable viral load, Caesarean section reduces vertical transmission to the same degree as documented previously for all women. Although the absolute risk reduction is very low, no study can show whether or not this is statistically significant and therefore women should be helped to make their individual choice. Mothers with HIV should not breastfeed in countries where formula milk is easily available, however highly active antiretroviral therapy administered to mothers or infants may reduce the risk of postnatal HIV transmission. SUMMARY: Counselling and assistance to conceive, modification of the therapeutic regimens and options about delivery have changed dramatically since the beginning of the HIV epidemic. Nowadays, women with HIV, similarly to uninfected women, can discuss with their doctors which therapeutic and treatment options would best fit their expectations of care. PMID- 15534443 TI - Female genital mutilation. AB - PURPOSE OF REVIEW: The purpose of this review is to provide an up to date account of recent papers and attitude on female genital mutilation in the past year. It is aimed at all professionals caring and supporting women/girls with female genital mutilation, and to identify gaps. RECENT FINDINGS: Given the multidisciplinary complexity of the practice of female genital mutilation, it is surprising how little empirical research (both quantitative and qualitative) exists. There is a paucity of rigorous research into its prevalence, its health consequences for those girls/women experiencing the practice, the understanding of professionals who have to address the issue, the implementation of existing legislation and the impact of programme interventions intended to change people's attitudes to, and the practice of, female genital mutilation. The overwhelming majority of literature focuses on the same topics, often drawing upon the same less-than-rigorous research data. SUMMARY: Worldwide, there are approximately 13 million refugees and asylum seekers, and human rights violations are seen as contributing factors to people fleeing their homeland. In the United States, those seeking asylum or refugee status are asked about their life experiences. African women are asked about ritual genital surgery, as it may be an indicator of their gynaecological, obstetric and sexual health. Health and social care professionals in host countries, in Europe, Australia, Canada, the USA and the UK, for example, are increasingly encountering this vulnerable client group in their practice and are finding that they are ill-prepared to deal with presenting complex health needs and challenges. PMID- 15534444 TI - Metformin in polycystic ovary syndrome. AB - PURPOSE OF REVIEW: Metformin has become an established treatment for women with polycystic ovary syndrome, although controversy remains as to how effective it is and in which populations it should be used. This review examines the recent literature in order to ascertain the evidence for the benefits and disadvantages of using metformin in women with polycystic ovary syndrome. RECENT FINDINGS: A Cochrane systematic review and metaanalysis examined the evidence for metformin in treating polycystic ovary syndrome. Since this was published there have been a number of new trials, some of which have been reasonably large involving participants from many different countries. SUMMARY: Evidence shows that metformin is effective in inducing ovulation, has some marginal benefit in improving aspects of the metabolic syndrome, improves objective measures of hirsutism, and seems to be effective in both obese and lean individuals. However, it has significant side effects, and the high levels of effectiveness that were reported by some early trials have not been replicated. Metformin should always be used as an adjuvant to general lifestyle improvements, and not as a replacement for increased exercise and improved diet. PMID- 15534445 TI - Levonorgestrel-releasing intrauterine system in the treatment of heavy menstrual bleeding. AB - PURPOSE OF REVIEW: Menorrhagia is a frequent reason for women to seek medical care and an increasingly common health problem. The levonorgestrel-releasing intrauterine system is an effective medical treatment for menorrhagia. Emerging clinical and research evidence suggests that this new treatment modality has major health benefits. RECENT FINDINGS: The levonorgestrel-releasing intrauterine system is a cost-effective treatment modality for menorrhagia. The quality of life improves significantly which is comparable to that gained with hysterectomy. The costs are about half those of hysterectomy after 5 years of follow-up. Unscheduled breakthrough bleeding is the most common side effect of the treatment. There are different new theories about the mechanism underlying this problem. Women with endometriosis or fibroids also benefit from this treatment. SUMMARY: Because menorrhagia is often a reason for seeking medical attention, it is important to consider the outcomes and costs to provide the most appropriate care. The levonorgestrel-releasing intrauterine system improves health-related quality of life significantly at relatively low cost. It is the most effective medical treatment for menorrhagia and comparable to surgical interventions. The system is not associated with serious complications. Although not all women are successfully treated, about 60% avoid hysterectomy and are satisfied with the treatment. Thus, the levonorgestrel-releasing intrauterine system should be the first line of treatment for heavy menstrual bleeding. PMID- 15534446 TI - Cervical screening in England and Wales: an update. AB - PURPOSE OF REVIEW: The National Health Service Cervical Screening Programme (NHSCSP) has played a major role in reducing the mortality from cervical cancer in England and Wales. However, the current system has numerous shortcomings and it is likely that its success has reached a plateau. In light of this, significant changes have recently been made to the programme. These alterations, as well as further potential developments, are considered here. RECENT FINDINGS: The aim of any change to the programme is to improve its sensitivity and specificity whilst reducing patient morbidity and maintaining cost-effectiveness. Alterations to NHSCSP guidelines include the replacement of the Papanicolau smear with liquid-based cytology, the referral for colposcopy of women with a single dyskaryotic cytology result and the commencement of screening at the later age of 25. These changes appear to be beneficial overall. The role of newer technologies in the programme is being clarified and it is likely that human papillomavirus testing will be incorporated in the near future. Progress is being made in the field of human papillomavirus vaccines, particularly prophylactic, which may go on to have the most profound impact on the incidence of cervical cancer. The disease is now largely a burden of the developing world, where the use of these technologies is considered. SUMMARY: Changes currently being instituted in the NHSCSP should go some way towards improving the service. The importance of increasing coverage rates, reducing patient waiting times and the associated anxiety must not be overlooked. PMID- 15534447 TI - Bibliography. Current world literature. Women's health. PMID- 15534450 TI - Editorial. PMID- 15534451 TI - Are we headed for a shortage of burn surgeons? AB - Newly trained burn surgeons are in high demand but low supply. The purpose of this study was to quantitate the current need for burn surgeons and to identify the need in the future. A questionnaire was sent to the 159 burn care facilities in North America listed in the 1999 to 2000 American Burn Association Directory. Of the 159 centers, 9 surveys were returned stating the center did not exist. Eighty-four (56%) of the remaining 150 centers responded. Twenty-nine percent of responding burn centers are currently attempting to recruit another burn surgeon, and an additional 38% anticipate needing to recruit one within the next 5 years. Of these, 89% anticipate difficulty finding another burn surgeon. This survey identifies a need for burn surgeons now and warns of a severe shortage in the immediate future. Current burn surgeons need to ensure that any interested trainee be mentored appropriately toward a career in burn surgery. PMID- 15534452 TI - Editorial. PMID- 15534453 TI - Guidelines for burn fellowships. PMID- 15534455 TI - Electrical injuries: a 20-year review. AB - Electrical injuries continue to present problems with devastating complications and long-term socioeconomic impact. The purpose of this study is to review one institution's experience with electrical injuries. From 1982 to 2002, there were 700 electric injury admissions. A computerized burn registry was used for data collection and analysis. Of these injuries, 263 were high voltage (> or =1000 V), 143 were low voltage (<1000 V), 277 were electric arc flash burns, and 17 were lightning injuries. Mortality was highest in the lightning strikes (17.6%) compared with the high voltage (5.3%) and low voltage (2.8%) injuries, and mortality was least in electric arc injuries without passage of current through the patient (1.1%). Complications were most common in the high-voltage group. Mean length of stay was longest in this group (18.9 +/- 1.4 days), and the patients in this group also required the most operations (3 +/- 0.2). Work related activity was responsible for the majority of these high-voltage injuries, with the most common occupations being linemen and electricians. These patients tended to be younger men in the prime of their working lives. Electrical injuries continue to make up an important subgroup of patients admitted to burn centers. High-voltage injuries in particular have far reaching social and economic impact largely because of the patient population at greatest risk, that is, younger men at the height of their earning potential. Injury prevention, although appropriate, remains difficult in this group because of occupation-related risk. PMID- 15534454 TI - The use of haloperidol and associated complications in the agitated, acutely ill pediatric burn patient. AB - The use of haloperidol to induce sedation and control agitation in the acutely ill adult patient has been well documented. There are few reports, however, of the use of this neuroleptic agent to control the severe delirium and agitation that may occur in critically ill pediatric patients or acute pediatric patients suffering from burn wounds. We assessed the effectiveness and safety of the use of haloperidol by completing a retrospective chart review of 855 acutely ill children treated consecutively during the period from April 1999 to May 2002, during which time 26 children received haloperidol. The safe use of haloperidol was assessed by documenting any adverse effects or reactions observed after the administration of the drug. Of patients given haloperidol, 23% had adverse effects. This result suggests that the use of haloperidol to treat the acutely agitated and delirious pediatric burn patients is fraught with a number of difficulties and is not completely safe and effective. PMID- 15534456 TI - Telemedicine follow-up of burns: lessons learned from the first thousand visits. AB - Telemedicine is an evolving technology that is used for health education, health care administration, and health care distribution. The potential benefits of telemedicine include a decrease in travel expenses, improved continuity of care, and increased access to specialized consultants, thus meeting the needs of patients, practitioners, and communities. Telemedicine has many evolving applications, including improved access to health care in medically underserved and rural areas. Regions Burn Center assessed the efficacy and efficiency of burn visits via telemedicine and identified the barriers and benefits specific to burn care. Information regarding travel costs and financial data were evaluated from a total of 1000 burn follow-up visits with 294 patients via telemedicine during a 5 year interval. Our results indicate that telemedicine burn visits are a cost effective clinical alternative for the patient. However, telemedicine can be a financial burden to health care systems and inefficient for health care providers. PMID- 15534457 TI - Circulating leptin and cortisol after burn injury: loss of diurnal pattern. AB - Leptin, a hormone involved in appetite and metabolic energy expenditure, could have a role in the reduced appetite and/or energy expenditure after burn injury. In this study, the diurnal pattern of circulating leptin concentrations was compared with body mass index (BMI), sex, glucose, insulin, and the diurnal cortisol rhythm in burn patients. Plasma samples were collected at 12:00 pm and 02:00 am from severely burned adults and children. Circulating leptin, insulin, and cortisol were measured by radioimmunoassay. Results were compared with previously published data from healthy control subjects. Overall, plasma leptin levels were lower in burn patients (5.7 +/- 1.2 ng/mL) compared with control subjects (10.5 +/- 1.7 ng/mL, P = .02). The normal nocturnal increase of circulating leptin concentrations observed in control subjects was completely absent in burn patients. Cortisol levels were higher in burn patients (20.4 +/- 1.0 mg/dL) than in control subjects (9.8 +/- 1.6 mg/dL, P < .0001) and the normal circadian decrease of circulating cortisol levels was markedly blunted in burn patients. Plasma cortisol did not correlate with circulating leptin levels. Plasma insulin and plasma glucose levels were significantly elevated in burn patients and the insulin:glucose ratio was dramatically increased compared with control subjects. Patients with burn injuries exhibited significantly decreased circulating leptin levels. This decrease may be the result of marked insulin resistance, as suggested by the elevated insulin to glucose ratio in burn patients. The loss of the diurnal pattern in burn patients is likely to result from the continuous nutritional supplementation. Because low leptin levels should induce appetite, burn-related anorexia is probably controlled by other regulatory systems. PMID- 15534459 TI - Sandal burns and their treatment in children. AB - Sandal is an ancient, primitive heating device that is still in use by both poor and rich people in mountain areas of Middle Asia. Sandal burn injuries are a serious health problem. Characteristics of sandal burns include not only skin injuries of various depths but also injuries to underlying tissues: subcutaneous fat, fasciae, muscles, and even bones. Sandal burns are characterized by such severe deep injuries because of a close contact of the body with live coals or woods. The main goal of this work was to present the most complete information about sandal burns and discuss the most effective methods of treatment for sandal burns. This treatment is used to accelerate the rejection of necrotic tissue, to prepare the wound for early autodermoplastic surgery, to decrease the postburn contractures/deformities, and also to shorten hospital stay for the patients. PMID- 15534460 TI - The medical use of a nonmedical product: melted tar. AB - The use of hot tar in industry is widespread, and burns caused by this agent remain a problem. Instead of being classified as chemical injuries, these burns compose a unique class of thermal injury because of the difference of mechanisms of injury and the difficulties in removing the agent from the skin. Tar burns usually are occupational in nature and are observed mostly in male patients. This report presents a female burn case that had occurred after she applied hot tar to her skin for the treatment purposes of pain relief. PMID- 15534461 TI - Gas fireplace contact burns in young children. AB - Contact burns from domestic appliances are common in young children. Recently, gas fireplaces have been recognized as a potential cause of contact burns in young children. We sought to quantify the frequency of gas fireplace contact burns in young children, to identify the etiology of contact, to describe the clinical presentation, and to describe clinical outcomes. Children with gas fireplace contact burn injuries presenting to The Hospital for Sick Children in Toronto (1999-2002) were identified using three data sources: the Canadian Hospitals Injury Reporting and Prevention Program Database, the Burn Unit Registry, and the Rehabilitation Services Database. Demographic, clinical, and outcomes data were collected on all children. During the 4-year study period, 27 children presented to the hospital because of a gas fireplace contact burn (approximately 9% of all contact burns). The median age of the children was 14 months (range, 8-36 months), with 16 boys (59%). Most children were burned in their own home. With regard to etiology, 10 children (37%) lost their balance near the fireplace, 2 (7%) walked too close to the glass front, and 8 (30%) touched the glass front out of curiosity. Almost half (44%) of the children burned the palms and digits of both hands. The median total burn surface area was 1% (range, 0.2-2.5%). In total, 30% of children were admitted to hospital, and 11% required skin grafts. All children had full wound closure after 4 to 43 days. Given the etiology of these burns (loss of balance or curiosity), passive prevention, such as barriers or changes in the composition of glass panels, may be the most effective approach to combat them. PMID- 15534462 TI - OT/PT. PMID- 15534463 TI - A survey of current rehabilitation trends for burn injuries to the head and neck. AB - Burn injuries to the face and neck present a unique challenge to the burn team and must be treated with considerable care and vigilance by the rehabilitation team to prevent potential contractures that may occur. Standard guidelines for treatment of the head and neck have not been established. This article presents the results of a comprehensive survey that examined the similarities and differences in the rehabilitation techniques used in various burn centers for the treatment of these injuries. The results obtained from the survey responses demonstrate several similarities in treatment throughout the continuum of care, revealing the potential for the development of a uniform rehabilitation protocol for the treatment of burn injuries to the face and neck. PMID- 15534464 TI - Pronator quadratus flap for burn reconstruction. AB - Full-thickness skin loss in the distal volar forearm presents a challenge to the burn surgeon. Two cases are presented in which a pronator quadratus muscle flap with split-thickness skin was used to cover the underlying tendons and nerves. The unique vascular perfusion makes this a useful and predictable transfer. PMID- 15534465 TI - Immunologists ponder glaucoma: a few inflammatory remarks. PMID- 15534466 TI - Incidence of secondary glaucoma in behcet disease. AB - PURPOSE: To determine the incidence of secondary glaucoma in Behcet disease. METHODS: A total of 230 eyes of 129 patients with Behcet disease, were examined in uveitis and glaucoma clinics of Ankara Social Security Eye Hospital between January 1997 and September 2002. The data from all patients were investigated both retrospectively and prospectively. RESULTS: The mean age of 129 patients was 34.2 +/- 7.4 years (range, 18 to 55 years). In 22 patients (17%), the disease was diagnosed on the basis of the ocular findings, while in the remaining 107 patients (83%), the period between the diagnosis of Behcet disease and the onset of the ocular symptoms was 23.3 +/- 17 months (range, 1 month to 5.3 years); 122 eyes (53%) had the episodes of acute recurrent iridocyclitis, while 108 eyes (47%) developed chronic posterior uveitis, including vitreitis, retinitis, vasculitis, or optic nerve involvement. Secondary glaucoma was diagnosed in 25 eyes (10.9%); 11 eyes (44%) with steroid or inflammation induced open angle glaucoma, 6 eyes (24%) with partial angle-closure glaucoma and peripheral anterior synechiae, 5 eyes (20%) with angle closure glaucoma, peripheral anterior synechiae, and pupil block and 3 eyes (12%) with neovascular glaucoma. The treatments included YAG-laser iridotomy in 5 eyes, diode-laser cyclodestruction in 3 eyes, primary trabeculectomies with mitomycin-c in 4 eyes, secondary trabeculectomies with mitomycin-c in 2 eyes, Ahmed valve implantations in 2 eyes, and cyclocryotherapy in 3 eyes. CONCLUSIONS: We suggest that secondary glaucoma is a common and serious complication of Behcet disease. It develops as a result of multiple factors, generally triggered by recurrent intraocular inflammation. Early recognition and treatment of these factors have vital importance to avoid the visual morbidity. PMID- 15534467 TI - Goniosurgery for glaucoma secondary to chronic anterior uveitis: prognostic factors and surgical technique. AB - PURPOSE: To describe the preoperative prognostic factors that correlate with the surgical success of goniosurgery for glaucoma complicating chronic anterior uveitis and to describe the surgical technique. METHODS: The medical records of 31 patients with 31 eyes with glaucoma secondary to chronic anterior uveitis for which 38 goniotomy procedures were performed were reviewed. Uveitis etiology, gender, age of onset of iritis, duration of iritis before recognition of glaucoma, duration of iritis and duration of glaucoma until initial goniosurgery, preoperative gonioscopic findings, lens status, surgical outcome, age at initial goniosurgery, duration of postoperative observation, lens status, preoperative and postoperative intraocular pressures, topical steroid, and glaucoma medication use were reviewed. Complete success was defined by an intraocular pressure (IOP) < or = 21 mm Hg, and qualified success as IOP < or = 21 mm Hg with use of glaucoma medications. The surgical technique used to perform the goniosurgery was reviewed. RESULTS: Overall surgical success was achieved in 22 (71%) of 31 eyes. The mean age at surgery was 10.2 +/- 4.2 years (range, 4-21 years). All but 8 patients were female. Mean follow-up interval was 10.3 +/- 6.4 years (range, 1.5 22 years). Significant correlation was found between outcome and age at initial surgery, lens status, duration of glaucoma before goniosurgery, duration of iritis before glaucoma surgery, and hours of peripheral anterior synechia (PAS). The mean number of clock hours of preoperative (PAS) was 0.5 hours in eyes with success versus 2.5 hours in eyes with failure. Age of onset of iritis, duration of iritis before recognition of glaucoma, trabecular meshwork pigmentation (TM), TM opacification, and circumferential ciliary body band narrowing did not correlate with surgical outcome. CONCLUSION: Goniosurgery is an effective treatment of glaucoma secondary to chronic anterior uveitis. The outcome of surgery was unfavorably influenced by older age, longer duration of glaucoma, and evidence of more advanced preoperative filtration angle abnormalities secondary to uveitis. Goniosurgery for this secondary glaucoma can be successfully performed utilizing the standard goniotomy technique. PMID- 15534468 TI - Trabeculectomy with mitomycin-C in uveitic glaucoma associated with Behcet disease. AB - PURPOSE: To determine the effect of intraoperative application of Mitomycin-C (MMC) with trabeculectomy in uveitic glaucoma associated with Behcet disease. MATERIALS AND METHODS: Twenty-six eyes of 26 patients with uveitic glaucoma associated with Behcet disease who underwent trabeculectomy with MMC between 1996 and 2001 were reviewed in this retrospective, noncomparative study. Trabeculectomy + MMC in concentration of 0.4 mg/mL for 3 minutes was performed to all patients. Main outcome measures were control of IOP, the number of antiglaucoma medications required to achieve the desired IOP, visual acuity and complications. The surgical success was defined as IOP less than 22 mm Hg and greater than 5 mm Hg without additional further glaucoma surgery or loss of light perception. RESULTS: The cumulative probability of success was 83.3% at 1 year, 76.2% at 2 years, 70% at 3 years, 66.7% at 4 years and 62.5% at 5 years after surgery. The mean follow-up was 40.0+/-18.0 months. At last follow-up 23% of the patients required no antiglaucoma medications. Best-corrected visual acuity improved or remained within two lines of preoperative visual acuity in 19 eyes (73.1%). Glaucomatous (1 eye 3.8%) and nonglaucomatous optic atrophy (3 eyes 11.5%) was the most frequent reason for visual decrease (total 4 eyes 15.2%). The most common complications were cataract formation in 6 eyes (23.1%), bleb leakage in 4 eyes (15.3%) and choroidal effusion in 3 eyes (11.5%). Phthisis bulbi was found in one (3.8%) patient. CONCLUSION: Trabeculectomy and intraoperative application of MMC appears to provide long term safety and effectiveness in uveitic glaucoma associated with Behcet disease. PMID- 15534469 TI - A pilot study of a system for grading of drainage blebs after glaucoma surgery. AB - PURPOSE: To develop and evaluate a novel bleb grading scheme for clinical and photographic evaluation. METHOD: A system for grading bleb photographs using widely applicable parameters was designed, and reference color photographs printed. A prospective masked agreement study was undertaken comparing slit lamp examination with mono and stereo photographs; 36 eyes of 28 patients with previous glaucoma surgery were graded according to defined parameters on a 1 to 10 scale clinically at the slit lamp by four ophthalmologists and two optometrists. Standardized stereo and mono photographs of the blebs were taken on the same day. The photographs were graded at least one week later in a masked fashion by the same observers, with grading of mono and stereo photographs also separated by one week. Analysis was performed to determine the variability and agreement between slit lamp results and photographic results, and to identify the presence of systematic bias. RESULTS: High levels of agreement were found between slit lamp and both stereo and mono photographs for vascularity indices, bleb wall thickness, and bleb elevation. Lower levels of agreement were found for the relative components of demarcated versus diffuse areas of the bleb, and for the total width of the bleb. The interquartile range for the median difference between slit lamp and photograph grading was -1.0 to 1.0 for all criteria except diffuse component (-2.0 to 2.0), and the median difference for all scores was 0.0. The median interobserver difference for all criteria was 0.0; the quartile range for all scores was between -0.5 and 1.0 except for diffuse component and width assessments whose quartiles fell in the -1.75 to 1.0 range. Examiners agreed with photographic grading within +/- 1 in more than 80% of gradings for vascularity and bleb height, within +/- 1 in more than 75% of gradings for bleb wall thickness, within +/- 2 in 61% of bleb width assessments, and +/- 2 in 59% of diffuse component. CONCLUSION: This bleb grading system is reproducible clinically and photographically. High levels of agreement between scores for photographs versus slit lamp examination were found for most categories, with good interobserver agreement for both photograph and slit lamp grading. Further refinement of scoring and reference photographs is required for optimization, especially for grading of bleb morphology. PMID- 15534470 TI - Incidence of glaucoma in patients with uveitis. AB - PURPOSE: To evaluate the incidence of glaucoma and elevation of intraocular pressure (IOP) in patients with inflammatory eye disease. METHODS: Retrospective review of medical records of 391 consecutive patients with uveitis attending a uveitis clinic of an academic Department of Ophthalmology from January 1999 to August 2002. Demographic, ocular and systemic variables were recorded. The diagnosis and treatment of uveitis were recorded. Uveitis was classified according to standard anatomic, etiological and clinical criteria. "Glaucoma" was defined as elevated IOP (>21 mm Hg) or glaucomatous optic nerve damage requiring medical and/or surgical anti-glaucoma treatment. Kaplan-Maier analysis and log rank tests were used to evaluate and compare the incidence of glaucoma. RESULTS: The incidence of glaucoma as defined above at 3 and 12 months after acute uveitis was 7.6%. In patients with chronic uveitis (n = 337), the incidence of glaucoma at 1 and 5 years was 6.5% and 11.1%, respectively. There was no statistically significant difference in the incidence of glaucoma between different types of uveitis, idiopathic versus non-idiopathic, and among anterior, intermediate, posterior and panuveitis. Visual loss occurred more frequently in patients with glaucoma than in patients without glaucoma. CONCLUSION: In patients with chronic inflammatory eye disease, the presence of glaucoma was associated with an increasing risk of visual loss. The incidence of glaucoma increased with time and was similar among the different types of uveitis. PMID- 15534471 TI - Novel MYOC gene mutation, Phe369Leu, in Japanese patients with primary open-angle glaucoma detected by denaturing high-performance liquid chromatography. AB - PURPOSE: To screen for mutations in the MYOC gene in Japanese patients with primary open-angle glaucoma (POAG) using denaturing high-performance liquid chromatography (DHPLC). PATIENTS AND METHODS: Blood samples were collected from 171 patients with POAG and 100 controls from seven institutions in Japan. For high-throughput analysis, seven exonic regions were amplified by polymerase chain reaction using DNA pooled from three patients; each DNA pool was then analyzed chromatographically. For analysis of a small number of samples, 7 exonic regions were amplified separately but simultaneously with annealing at 58 degrees C in each patient and then chromatographed, using 7 wells of the same 96-well plate per sample. When chromatographic patterns were abnormal by either method, the PCR products of the individual samples were sequenced. RESULTS: Four glaucoma-causing mutations were identified in five POAG patients (2.9%). One missense mutation, Phe369Leu, is new; and three others, Ile360Asn, Ala363Thr, and Thr448Pro, have been reported in Japanese patients. Phe369Leu was associated with adult onset POAG. CONCLUSIONS: Mutations in the MYOC gene were demonstrated chromatographically in 2.9% of our Japanese POAG patients. The use of pooled DNAs with DHPLC analysis is a time- and labor-saving technique. All mutations detected appear to be specific to Japanese patients. PMID- 15534472 TI - Ultrasound biomicroscopy of intrascleral lake after viscocanalostomy and cataract surgery. AB - PURPOSE: To assess anatomic changes at the sclerectomy site after combined viscocanalostomy and cataract surgery associated with POAG or OH and cataract. METHODS: In a prospective, noncomparative, longitudinal study, we performed ultrasound biomicroscopy (UBM) of the sclerectomy site in 84 eyes at 3, 6, and 12 months after the surgery. Four parameters in UBM findings (area, radial longitudinal length, height of intrascleral space, and meridional length of window) were measured and the IOP level, the number of antiglaucoma medications, and bleb formation were observed for 12 months. RESULTS: Intraocular pressure decreased from 19.8 +/- 3.5 (+/- SD) mm Hg preoperatively to 15.5 +/- 3.0 mm Hg 12 months postoperatively (P < 0.0001). Blebs were formed in 2 of 84 eyes (2.4%) at 1 month, and 1 of 84 eyes (1.2%) after 3 months. The IOP decreased at 3, 6, and 12 months postoperatively by 5.8, 4.8, and 4.3 mm Hg, respectively (P = 0.0016). A significant difference was observed between 3 months and 12 months in area (0.17 and 0.13 mm, respectively, P = 0.0127 using the Bonferroni/Dunn test), longitudinal length (0.89 and 0.72 mm, P = 0.0059), and height (0.21 and 0.15 mm, P < 0.0001), but not in meridional length (0.41 and 0.33 mm). The lake was present in 66.7% of cases at 12 months. CONCLUSION: Combined viscocanalostomy and cataract surgery lowered IOP without bleb formation. Postoperatively, the size of the lake and IOP decreased, suggesting parallel reduction of the two. The lake was undetected ultrasonographically in one third of the cases 1 year postoperatively. PMID- 15534473 TI - Evaluation of scanning resolution on retinal nerve fiber layer measurement using optical coherence tomography in normal and glaucomatous eyes. AB - PURPOSE: To evaluate the effect of varying the scanning resolution of optical coherence tomography (OCT) retinal nerve fiber layer (RNFL) measurement on diagnostic sensitivity and functional correlation in glaucoma. PATIENTS AND METHODS: 314 eyes from 182 subjects including 107 normal eyes, 83 glaucoma suspect eyes, and 124 glaucoma eyes were included in this cross-sectional study. Standard automated perimetry and OCT measurement of RNFL thickness were performed. Each individual underwent two scanning protocols: (1) fast RNFL thickness (3.4) scan (with resolution of 256 scan points) and (2) RNFL thickness (3.4) scan (with resolution of 512 scan points). RNFL thickness was compared among the groups. Diagnostic sensitivity was evaluated with Receiver Operating Characteristic (ROC) Curve. Relationship between RNFL thickness and visual field mean deviation was examined using linear regression analysis. RESULTS: Measured RNFL thickness using fast RNFL thickness (3.4) scan was significantly higher compared with RNFL thickness (3.4) scan in average, superior, nasal and inferior RNFL in all diagnostic groups. Comparing normal and glaucoma groups, RNFL thickness (3.4) scan produced the largest area under the ROC curve (0.912) based on average RNFL thickness. A stronger correlation between average RNFL and visual field mean deviation was found in RNFL thickness (3.4) scan (R = 0.75, R = 0.56). CONCLUSIONS: Higher resolution RNFL scan provides better diagnostic sensitivity in glaucoma detection and a stronger correlation with visual function. PMID- 15534474 TI - Altered transcripts expression of matrix metalloproteinases and their tissue inhibitors in tenon capsule of patients with glaucoma. AB - PURPOSE: To determine the transcripts expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) in Tenon capsule of patients with primary glaucoma and non-glaucomatous patients who serve as the control. PATIENTS AND METHODS: Specimens of Tenon capsule were obtained intraoperatively and evaluated with the reverse transcriptase-polymerase chain reaction method. The transcripts levels of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 relative to that of glyceraldehyde phosphate dehydrogenase were determined. RESULTS: The transcripts levels of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 were not correlated with age in the control group. There were differences in the transcripts expression of MMP-2 (P < 0.001), MMP-9 (P = 0.017), TIMP-1 (P < 0.001), and TIMP-2 (P = 0.001) among the control, primary open angle glaucoma, and primary angle-closure glaucoma groups (Kruscal-Wallis H test). Comparing with the controls, open angle glaucoma had decreased transcripts levels of MMP-2 (P = 0.001) and TIMP-1 (P = 0.006) while angle-closure glaucoma had decreased transcripts levels of MMP-2 (P < 0.001), MMP-9 (P = 0.004), TIMP-1 (P < 0.001), and TIMP-2 (P < 0.001) (Mann Whitney U tests). CONCLUSION: The transcripts expression of certain MMPs and TIMPs is altered in Tenon capsule of glaucoma patients, which might result from long-term application of topical glaucoma medications. PMID- 15534475 TI - Investigation of the association between OPA1 polymorphisms and normal-tension glaucoma in Korea. AB - PURPOSE: OPA1, the gene responsible for autosomal dominant optic atrophy, represents a good candidate gene for normal-tension glaucoma (NTG). Single nucleotide polymorphisms on intervening sequence (IVS) 8 of the OPA1 gene (IVS8+4C>T; +32T>C) were recently found to be strongly associated with NTG in a Caucasian population. We investigated whether these polymorphisms in the OPA1 gene were associated with NTG in Korea. PATIENTS AND METHODS: Sixty-five Korean NTG patients and 101 healthy Korean subjects were enrolled. DNA from peripheral blood leukocytes was extracted and the genotypes of two polymorphisms (IVS8+4C>T; +32T>C) in the OPA1 gene were determined using the restriction fragment length polymorphism method. The genotype and allele frequencies of two polymorphism in patients with NTG and normal controls were compared using the Fisher exact test and the chi test. Frequencies of haplotypes and haplotypes groups were also analyzed to assess the combined effect of two polymorphisms. RESULTS: The frequencies of the CT genotype of IVS8+4C>T, CC genotype of IVS8+32T>C, and TT genotype of IVS8+32T>C were not significantly different between NTG patients and controls (4.6% versus 0.0%, P = 0.058 by the Fisher exact test; 10.8% versus 4.0%, P = 0.11 by the Fisher exact test; 61.5% versus 67.3%, P = 0.45 by the chi test, respectively). Any haplotype or haplotype group of IVS8+4C>T and IVS8+32T>C was not associated with NTG, and the C allele of IVS8+32T>C was not a significant modifier of IVS8+4C>T. CONCLUSIONS: There were no significant associations between IVS8+4C>T; +32T>C polymorphisms and NTG in the Korean population. These results do not support the results in Caucasians and indicate that ethnic differences may exist in the association between polymorphisms in the OPA1 gene and NTG. PMID- 15534476 TI - Neuronal nitric oxide synthase (nNOS) positive retinal amacrine cells are altered in the DBA/2NNia mouse, a murine model for angle-closure glaucoma. AB - PURPOSE: To characterize retinal amacrine cell changes in eyes of DBA/2NNia (DBA) mice that develop an inherited angle-closure glaucoma. METHODS: DBA and non glaucomatous C57BL/6J mice of different age groups (2 to 23 months of age) were studied and compared. Morphologic investigations included NADPH-diaphorase staining of retinal whole mounts and fluorescence immunohistochemistry of cryosections with antibodies against neuronal nitric oxide synthase (nNOS), tyrosin hydroxylase (TH), gamma aminobutyric acid (GABA), and vesicular acetylcholine transporter (VAChT). RESULTS: Immunohistochemistry of amacrine cell subpopulations in the retinae of DBA mice revealed no significant staining differences in the two mouse strains at all ages using antibodies against TH, GABA, and VAChT. However, staining with nNOS and NADPH diaphorase revealed significant differences between the DBA strain and the C57BL/6J mice. With the onset of elevated IOP and glaucoma beginning at around 6 months in the DBA mice, the total number of NOS positive amacrine cells continuously decreased from 1000 cells at 6 months of age down to 480 cells in animals older than 20 months of age, but did not decline in age-matched C57 mouse retinas. CONCLUSION: We previously described a parafoveal loss of nNOS positive amacrine cells in the monkey glaucoma model. The fact that there is also a significant decrease of nNOS amacrine cells in the glaucomatous mouse eye indicates a specific response of nNOS positive amacrine cells in glaucomatous retinopathy. PMID- 15534477 TI - Deep sclerectomy versus punch trabeculectomy with or without phacoemulsification: a randomized clinical trial. AB - PURPOSE: To compare the efficacy of non-penetrating deep sclerectomy without implant with Crozafon-De Laage punch trabeculectomy, and to evaluate the effect of simultaneous temporal approach phacoemulsification on both techniques. PATIENTS AND METHODS: SETTING: Department of Ophthalmology of the University of Palermo. DESIGN: Prospective randomized clinical trial. Patients and intervention procedures: Sixty-five patients (65 eyes) with primary open angle glaucoma (POAG) or pseudoexfoliative glaucoma (PEXG): 32 eyes underwent non-penetrating deep sclerectomy (NPDS), 17 as single procedure and 15 combined with phacoemulsification (phaco-NPDS), and 33 eyes underwent punch trabeculectomy (PT), 18 single and 15 with phaco (phaco-PT). The patients were randomly assigned to each procedure. No adjuvants, such as Nd: YAG laser goniopuncture, laser suturelysis, and antimetabolites were used. MAIN OUTCOME MEASURES: Postoperative complications, number of antiglaucoma medications, and IOP level were checked at each control. Complete success indicated the achievement of the target IOP without antiglaucoma medications, while qualified success indicated the same goal with or without medications. These categories were assessed at two target IOP levels, namely < or =21 mm Hg and < or =17 mm Hg in all four groups. RESULTS: The mean follow-up period was 22.5 +/- 2.5 months. The mean preoperative IOP was 30.2 mm Hg in NPDS eyes, 26.8 in phaco-NPDS eyes, 32.1 in PT eyes, and 27.0 in phaco PT ones, without significant intergroup difference. At the end point the mean IOP was 17.7 +/- 0.8, 15.7 +/- 0.9, 14.2 +/- 1.1, and 13.8 +/- 1.1 mm Hg respectively with postoperative IOP significantly lower (P = 0.005) than preoperative IOP in all groups. No difference was observed among groups at any observation time when simple and combined surgery were compared. Significant difference at the end point was found between NPDS and PT (P = 0.030). As for complete and qualified success with a < or =21 and < or =17 mm Hg target IOP no significant differences were noticed in all groups. Among postoperative complications, hypotony was significantly more frequent in both PT groups when compared with the NPDS groups. The same was true, but relating only to the single procedures, for shallow anterior chamber and choroidal detachment. The Kaplan-Meier cumulative survival curves relating to the qualified success rate in the four surgical groups for a < or =21 mm Hg target IOP (log rank, P = 0.564) and for a < or =17 mm Hg target IOP (log rank, P = 0.591) showed no significant intergroup differences. When the < or =21 mm Hg target IOP was considered, a mild positive trend in combined procedures (both phaco-NPDS and phaco-PT) was found in comparison with simple procedures. At lower IOP target (ie, < or =17 mm Hg) a better trend was found in favor of simple or combined PT procedure. CONCLUSIONS: Both techniques, NPDS and PT, without enhancements (ie, implants or antimetabolites) control IOP efficaciously at our end point. Phacoemulsification combined with penetrating and non-penetrating procedures does not seem to interfere with final results. When a lower target IOP and probability of success over time are considered, PT, single or combined, exhibits a better trend. PT, therefore, could be more suitable for higher IOP levels or longer life expectancies. PMID- 15534478 TI - Comparison of preoperative conjunctival bacterial flora in patients undergoing glaucoma or cataract surgery. AB - PURPOSE: To assess differences in conjunctival bacterial flora between patients undergoing glaucoma and cataract surgery. PATIENTS AND METHODS: A prospective study comparing conjunctival bacterial cultures obtained from 339 patients undergoing either cataract (n = 258) or glaucoma (n = 81) surgery. All cultures were acquired during the preoperative visit, approximately three to seven days prior to surgery. The culture samples were inoculated onto blood and chocolate agar, as well as blood culture broth media. All bacterial isolates were identified and statistical analyses were performed to determine if there were differences in flora between the eyes undergoing cataract versus glaucoma surgery. RESULTS: Two hundred fifteen of 258 eyes (83%) undergoing cataract surgery were found to have positive bacterial growth, compared with 62 of 81 eyes (77%) of those undergoing glaucoma surgery (P = 0.2246). Coagulase-negative Staphylococci, the most common bacterial isolate, was cultured from 167 eyes (65%) in the cataract group and 42 (52%) in the glaucoma group (P = 0.0514). Among all bacterial isolates, only Corynebacterium species was found to be statistically different between the two patient groups with 92 (36%) and 11 (14%) eyes testing positive in the cataract and glaucoma groups, respectively (P = 0.0003). CONCLUSIONS: There was no statistically significant difference in the proportion of conjunctival culture samples testing positive for bacterial growth in eyes undergoing glaucoma surgery compared with those undergoing cataract surgery. Glaucoma medications, or their preservatives, do not appear to significantly alter conjunctival flora. Techniques used for endophthalmitis prophylaxis prior to cataract surgery are likely appropriate for glaucoma surgery as well. PMID- 15534479 TI - Contact-topical plus intracameral lidocaine versus peribulbar anesthesia in combined surgery: a randomized clinical trial. AB - PURPOSE: To compare the efficiency and safety of contact-topical anesthesia versus peribulbar injection anesthesia for phacotrabeculectomy. MATERIALS AND METHODS: A total of 80 patients undergoing combined cataract and glaucoma surgery were randomly allocated to receive either contact or peribulbar anesthesia. No systemic sedatives were used in either group. Patients were asked to rate their pain level on a 5-point scale for 4 periods: during administration of the anesthetic agent; during surgery; immediately after surgery; and 24 hours postoperatively, while the surgeon recorded his subjective assessment of ease of surgery using a standardized template. The patients' general condition during surgery, as well as the results and short-term complications, were assessed. RESULTS: The injected anesthesia group showed higher rates of discomfort and pain, and 37 patients reported pain ranging from mild to severe during anesthetic administration. The difference between groups was statistically significant (P < 0.001). During surgery, there were no differences in vital signs, patients' subjective pain evaluation, or surgeon stress. We found no differences between pain rates after surgery. Complications included prolonged chemosis, and we also noted that conjunctival hemorrhage occurred more frequently in the peribulbar group than in the contact anesthesia group. CONCLUSION: Both anesthetic methods provide high levels of pain control without additional sedation during surgery. The use of contact-topical anesthesia avoids pain and reduces the possibility of complications during administration of anesthetics. PMID- 15534480 TI - Evaluation and comparison of indentation ultrasound biomicroscopy gonioscopy in relative pupillary block, peripheral anterior synechia, and plateau iris configuration. AB - PURPOSE: To evaluate and compare the findings and changes of the anterior chamber angle configuration with indentation ultrasound biomicroscopy (UBM) gonioscopy in relative pupillary block (RPB), peripheral anterior synechia (PAS), and plateau iris configuration (PIC). METHODS: This study included 73 eyes of 52 patients with RPB (n = 26), PAS (n = 21), or PIC (n = 26). First, a conventional UBM scan was performed using a normal size standard eye cup before indentation. Then, for indentation UBM gonioscopy, scans were performed using a new eye cup that we designed. For evaluation of the angle, angle opening distance 500 and angle recess area were recorded and evaluated with regard to the effect of expansion on the anterior chamber angle. RESULTS: Indentation UBM gonioscopy showed the characteristic images in each of the eyes. The angle of all examined eyes was significantly widened with indentation (P < 0.01). The angle changes in eyes with RPB were significantly greater than in eyes with PAS or PIC (P < 0.01). CONCLUSION: Indentation UBM gonioscopy is a very useful method for observing the angle and diagnosis of RPB, PAS, and PIC. PMID- 15534481 TI - Inferonasal placement of aqueous shunts. PMID- 15534485 TI - The generation and anti-myeloma activity of a chimeric anti-CD54 antibody, cUV3. AB - Despite new treatment options, including autologous and allogeneic stem cell transplants, multiple myeloma remains an incurable disease. The authors developed and characterized a murine anti-human ICAM-1 (CD54) monoclonal antibody, UV3, which is highly effective in SCID mice with advanced human myeloma xenografts (SCID/ARH-77). To improve the effector functions and pharmacokinetic parameters and to reduce its immunogenicity in humans, the authors engineered this monoclonal antibody into a mouse/human IgG1kappa chimeric (c) antibody, cUV3. Following coexpression and purification of the genetically spliced heavy and light chain constructs, the authors compared cUV3 and UV3 in various in vitro assays, including relative cell-binding affinities and effector functions, namely antibody-dependent cellular cytotoxicity and complement-dependent cytotoxicity. The authors compared their in vivo retention times and biodistribution patterns in normal mice. In each assay, the authors found that cUV3 was essentially equivalent to UV3. Finally, these antibodies were tested in a SCID/ARH-77 model of advanced myeloma, with daily treatments of 4 mug/g for 4 consecutive days commencing 14 days after tumor cell inoculation. cUV3 was at least as effective as UV3; 40% and 20% of the mice, respectively, were cured, with no sign of disease at day 150. The authors intend to evaluate the efficacy of cUV3 further in SCID/ARH-77 mice using other doses and dosing schedules to try to improve the cure rate. Eventually, they hope to test the efficacy of cUV3 in patients with multiple myeloma. PMID- 15534486 TI - Competition among peptides in melanoma vaccines for binding to MHC molecules. AB - The effectiveness of peptide-based cancer vaccines depends on the ability of peptides to bind to MHC molecules on the surface of antigen-presenting cells, where they reconstitute epitopes for cytotoxic T lymphocytes (CTLs). Multivalent vaccines have advantages over single-peptide vaccines; however, peptides may compete for binding to the same MHC molecules. In particular, it is possible that peptides with high affinity for MHC molecules prevent the binding of lower affinity peptides. However, only small numbers of peptide/MHC complexes per cell are required for CTL recognition. Thus, the authors hypothesized that competition of peptides for MHC binding would not significantly reduce CTL recognition of individual peptides within a multiple-peptide mixture, and this hypothesis was tested by a series of experiments performed in vitro. In multiple experiments, two peptides with different affinities for HLA-A*0201 molecules were mixed at various concentrations and pulsed onto HLA-A2 cells, which were then evaluated for susceptibility to lysis by HLA-A*0201-restricted CTLs. CTL recognition of the melanoma peptides gp100(154-162) (KTWGQYWQV), gp100(280-288) (YLEPGPVTA), and tyrosinase(369-377D) (YMDGTMSQV) was maintained even when target cells were co pulsed with equimolar concentrations of peptides with comparable or higher affinity for HLA-A2. In some cases, CTL recognition was maintained even when the higher-affinity peptide was present at concentrations several orders of magnitude higher than the target peptide. In addition, CTLs generated by in vitro stimulation with a peptide mixture developed reactivity to three different peptides, at a level comparable to that obtained by stimulation with each individual peptide separately. These data suggest that CTLs can respond to multiple peptides presented on the same antigen-presenting cells and justify further investigation, in clinical trials, of multiple-peptide cancer vaccines. PMID- 15534487 TI - Anti-tumor effect of an intratumoral administration of dendritic cells in combination with TS-1, an oral fluoropyrimidine anti-cancer drug, and OK-432, a streptococcal immunopotentiator: involvement of toll-like receptor 4. AB - The authors investigated the in vivo anti-tumor effect of intratumoral administration of bone marrow-derived dendritic cells (DCs) after chemotherapy using an oral fluoropyrimidine anti-cancer drug TS-1, and followed by immunotherapeutic agent OK-432, in two syngeneic tumor-bearing mouse models. Both in Meth-A fibrosarcoma-bearing BALB/c mice and in SCCVII-bearing C3H/HeN mice, 1 week of oral administration of TS-1 effected partial eradication of established tumors. Intratumoral injection of DCs and OK-432 caused only slight inhibition of the tumor growth. However, TS-1 administration followed by DCs and OK-432 resulted in a marked inhibition in the tumor growth and also contributed to a greater prolongation of survival. By the injection of DCs and OK-432 after TS-1 administration, a significant infiltration of immune cells, especially CD8+ T cells, was observed. Furthermore, the cytotoxic activities of tumor-infiltrating lymphocytes and draining lymph node cells against inoculated tumor cells were significantly increased by the therapy, while activities against nonspecific target cells were not. Cytotoxic memory T cells were also induced; the main effectors were MHC class I-restricted, CD8+ T cells. The same therapy was also applied to SCCVII-bearing C3H/HeJ mice in which the Toll-like receptor (TLR) 4 is mutated and its function impaired; no immunotherapeutic effect was observed in the TLR4-deficient mouse model. These findings suggest that the local DC therapy in combination with TS-1 and OK-432 may be a useful strategy for the treatment of solid tumors, and that TLR4 signaling is involved in the success of this therapy. PMID- 15534488 TI - Active specific immunotherapy of melanoma with a GM3 ganglioside-based vaccine: a report on safety and immunogenicity. AB - A novel cancer vaccine was obtained by combining GM3 ganglioside with Neisseria meningitidis outer membrane protein complex to obtain very-small-size proteoliposomes (GM3/VSSP). The authors report the results of a phase 1 study of intramuscular administration of GM3/VSSP/Montanide ISA 51 to patients with metastatic melanoma. Twenty-six patients were included in three dose-level cohorts of 120, 240, and 360 mug. The first five doses (induction phase) were given at 2-week intervals, and the remaining four doses were given monthly. Patients were evaluated for dose-related toxicities and antitumor effects. In addition, serum and peripheral blood mononuclear cells were obtained at baseline and throughout treatment to evaluate humoral and cellular immune responses. One episode of severe hypotension and fever was observed in a patient included at the highest dose level. Other toxicities consisted of local reactions at the site of injection and mild fever and chills. Five doses of GM3/VSSP induced an anti-GM3 IgM response in 44% of patients. Serum reactivity was also observed against melanoma cell lines and tumor biopsies. GM3/VSSP was shown to induce very strong in vitro IFNgamma secretion in all evaluated melanoma patients. Furthermore, in one patient IFNgamma secretion was shown to be GM3-specific. A 62% reduction of a mediastinal mass was documented in one patient (partial response), while a second patient benefited from initial disease stabilization followed by tumor reduction in nonmeasurable soft tissue lesions accompanied by vitiligo. PMID- 15534489 TI - Vaccination of glioma patients with fusions of dendritic and glioma cells and recombinant human interleukin 12. AB - Despite aggressive treatment, the median survival of patients with high-grade malignant astrocytoma is about 1 year. The authors investigated the safety and clinical response to immunotherapy using fusions of dendritic and glioma cells combined with recombinant human interleukin 12 (rhIL-12) for the treatment of malignant glioma. Fifteen patients with malignant glioma participated in this study. Dendritic cells were generated from peripheral blood. Cultured autologous glioma cells were established from surgical specimens in each case. Fusion cells were prepared from dendritic and glioma cells using polyethylene glycol. All patients received fusion cells intradermally on day 1. rhIL-12 was injected subcutaneously at the same site on days 3 and 7. Response to the treatment was evaluated by clinical observations and radiologic findings. No serious adverse effects were observed. In four patients, magnetic resonance imaging showed a greater than 50% reduction in tumor size. One patient had a mixed response. These results show that administration of fusion cells and rhIL-12 safely induces clinical antitumor effects in some patients with malignant glioma. PMID- 15534490 TI - Induction of systemic TH1-like innate immunity in normal volunteers following subcutaneous but not intravenous administration of CPG 7909, a synthetic B-class CpG oligodeoxynucleotide TLR9 agonist. AB - Subcutaneous injection of normal human volunteers with a B-class CpG oligodeoxynucleotide (ODN) TLR9 agonist, CPG 7909, induced a TH1-like pattern of systemic innate immune activation manifested by expression of IL-6, IL-12p40, IFN alpha, and IFN-inducible chemokines. Serum IP-10 was found to be the most sensitive assay for subcutaneous CPG 7909 stimulation; its level was significantly increased in all subjects at all dose levels, including the lowest tested dose of just 0.0025 mg/kg. This pattern of chemokine and cytokine induction was markedly different from that previously reported to be induced by TLR9 stimulation in rodents, most likely reflecting species-specific differences in the cell types expressing TLR9. Subcutaneous CPG 7909 injection induced transient shifts in blood neutrophils, lymphocytes, and monocytes, consistent with the increased chemokine expression. Levels of acute phase reactants such as C-reactive protein were also increased. A second subcutaneous CPG 7909 injection administered 2 weeks after the first elicited similar immune responses, showing little or no tolerance to the effects of repeated in vivo TLR9 stimulation. Subjects developed dose-dependent transient injection site reactions and flu-like symptoms but otherwise tolerated injection well, with no evidence of organ toxicity or systemic autoimmunity. The activation of innate immunity was dependent on the route of ODN administration, since intravenous injection caused no such effects. These studies indicate that in vivo activation of TLR9 by subcutaneous administration of CPG 7909 could be a well-tolerated immunotherapeutic approach for induction of TH1 innate immune activation. PMID- 15534491 TI - Immunization of HLA-A*0201 and/or HLA-DPbeta1*04 patients with metastatic melanoma using epitopes from the NY-ESO-1 antigen. AB - HLA class I-restricted peptides are often used in peptide vaccine regimens. There is strong evidence that many of these peptides can generate specific CD8 T-cell responses in vivo; however, only occasional objective clinical responses have been reported. To test whether provision of "help" would enhance antitumor immunity, the authors initiated a clinical trial in which patients with metastatic melanoma were immunized against the NY-ESO-1 tumor antigen, using an HLA-A2-restricted peptide (ESO-1:165V), an HLA-DP4-restricted peptide (NY-ESO 1:161-180), or both peptides given concomitantly. The first cohorts received only ESO-1:165V, using three vaccination schedules. Immunologically, most patients developed immune responses to the HLA-A2-restricted native ESO-1 epitope after vaccination. Peptide vaccine given daily for 4 days appeared to induce immunologic responses more rapidly than if given once a week or once every 3 weeks. In contrast, vaccination using the NY-ESO-1:161-180 peptide induced immune responses in only a few patients. Clinically, one patient who received NY-ESO 1:161-180 peptide alone had a partial response lasing 12 months. Concomitant vaccination with the HLA class II-restricted peptide did not alter the immune response to the HLA class I-restricted peptide form NY-ESO-1. However, vaccination with the HLA-A2-restricted epitope generated primarily T cells that did not recognize tumor after in vitro sensitization. This result raises questions about the use of synthetic peptides derived from NY-ESO-1 as a sole form of immunization. PMID- 15534492 TI - Cytotoxic T lymphocyte-associated antigen 4 blockade in patients with metastatic melanoma: a new cause of uveitis. AB - Cytotoxic T Lymphocyte-associated antigen 4 (CTLA-4) is an important costimultory receptor expressed on activated T cells. CTLA-4 blockade using a monoclonal antibody (mAb) in conjunction with tumor vaccines has improved tumor responses in animal models and enhanced numerous models of T cell-associated autoimmune diseases. Two patients with stage IV metastatic melanoma vaccinated with the gp 100 melanocyte/melanoma differentiation antigen either before or during anti-CTLA 4 mAb therapy developed uveitis. This is the first report of autoimmune disease involving the eye in patients treated with anti-CTLA-4 mAb. This suggests that CTLA-4 is an important regulatory molecule for maintenance of tolerance to melanosomal antigens and prevention of uveitis. PMID- 15534493 TI - Respiratory protection for mold remediation. PMID- 15534494 TI - Occupational asthma and rhinitis caused by milk proteins. PMID- 15534495 TI - Preplacement nerve testing for carpal tunnel syndrome. PMID- 15534497 TI - Factors affecting the frequency of value-focused health activities and policies by employers. AB - OBJECTIVE: We sought to gather employer perspectives about value-focused activities (VFAs), intentions to make decisions based on value, and other factors affecting decisions. METHODS: Health decision-makers (n = 174), both American College of Occupational and Environmental Medicine members and corporate HR/benefits directors, responded to an Internet-based questionnaire. RESULTS: Of a total of 32 listed VFAs, companies reported, on average, performing 5.2 activities currently and considering 2.6. Twenty-five percent of companies reported doing eight or more. The most common VFAs were providing access to flu shots, centers of excellence, and wellness programs. Greater access to detailed outcome data was associated with doing more VFAs, as was greater accountability for absence, disability, and productivity outcomes. CONCLUSIONS: Employers vary widely in the number of VFAs in which they participate. Decision-makers with more information about, and accountability for, value outcomes reported doing more VFAs. PMID- 15534498 TI - Do subject characteristics modify the effects of particulate air pollution on daily mortality among the elderly? AB - OBJECTIVE: Studies have reported associations between mortality and air pollution, but questions subsist on the identification of susceptible subgroups in the population. We studied individual characteristics that modify the relationship between particulate air pollution and mortality among elderly. METHOD: We examined 527 nonaccidental deaths (197 cardiorespiratory deaths) among the 1469 subjects from the Personnes Agees QUID cohort in Bordeaux between 1988 and 1997. Air pollution was measured as black smoke by urban monitoring background stations. We used a case crossover approach and calculated odds ratio by conditional logistic regression models. RESULTS: We observed associations between the third lag day and cardiorespiratory mortality for an increase of 10 microg/m3 of black smoke (odds ratio = 1.30, 95% confidence interval: 1.01-1.68). CONCLUSIONS: Our results provide insight into factors possibly conferring susceptibility to the acute effect of urban air pollution. PMID- 15534499 TI - Reliability and validity of the Stanford Presenteeism Scale. AB - OBJECTIVE: This study reports the reliability and validity of the 13-item Stanford Presenteeism Scale (SPS). The SPS differs from similar scales by focusing on knowledge-based and production-based workers. METHODS: Data were obtained from administrative and medical claims databases and from a survey that incorporated the SPS, SF-36, and the Work Limitations Questionnaire. RESULTS: Sixty-three percent (7797) of employees responded. Cronbach's alpha (0.83) indicates adequate reliability. Factor analysis identified two underlying factors, "completing work" and "avoiding distraction." Knowledge-based workers load on "completing work" (alpha = 0.97), whereas production-based workers load on "avoiding distraction" (alpha = 0.98). There were significant and positive relationships between the SPS, SF-36, and Work Limitations Questionnaire. CONCLUSIONS: The SPS demonstrates a high degree of reliability and validity and may be ideal for employers who seek a single scale to measure health-related productivity in a diverse employee population. PMID- 15534500 TI - How do job characteristics, family situation, domestic work, and lifestyle factors relate to sickness absence? A study based on Sweden Post. AB - OBJECTIVE: We sought to determine how do working life and private life characteristics relate to sickness absence. METHODS: Questionnaire data for 1557 female and 1913 male employees were related to registered sickness absence by multivariate analyses. RESULTS: Apart from health problems, clear associations with sickness absence were observed for complaints attributed to heavy, arduous work, and sickness presenteeism. The use of tranquilizers, occurrence of bullying, and the existence of a high total workload from paid and unpaid work were associated with sickness absence in women. In men, the use of alcohol as sedative, anxiety of reorganization, not holding a supervisor position, adverse life events, and divorce were related with sickness absence. CONCLUSIONS: Our study identified different areas at work and outside work for women and men that could be targets for actions aiming at preventing high sickness absence. PMID- 15534501 TI - Shift work, job stress, and late fetal loss: The National Birth Cohort in Denmark. AB - OBJECTIVE: The Danish National Birth Cohort (DNBC) was used to examine whether shift work or job stress correlate with late fetal loss. METHODS: We identified 33,694 pregnancies of daytime workers and 8,075 pregnancies of shift workers in women recruited to the DNBC between 1998 and 2001. Pregnancy outcomes were obtained by linkages to the national registers. Hazard ratios of fetal loss were calculated by using Cox regressions with left truncation. RESULTS: Fixed night work was associated with fetal loss (hazard ratio = 1.85, 95% confidence interval = 1.00-3.42). No high risk of fetal loss was seen for other types of shift work. Job stress, as measured in our study, was not associated with fetal loss. CONCLUSIONS: Our results suggest that fixed night work during pregnancy increases the risk of late fetal loss. PMID- 15534503 TI - Violence in healthcare facilities: lessons from the Veterans Health Administration. AB - GOALS: The authors examined assault frequency and risk factors in health care. METHODS: The authors conducted a cross-sectional questionnaire survey in 142 hospitals. Analyses are presented at the level of the individual and aggregated by facility. RESULTS: Thirteen percent of employees described at least 1 assault in the last year; the proportion assaulted per facility ranged from 1% to 26%. Patients were the most common assaulters. Working in geriatrics, mental health, and rehabilitation or in nursing represented a high risk for assault. Hours of work and work patterns represented major risk factors for assault, as were higher measures of organizational stress. The penetration of training in alternate dispute resolution strategies was associated with lower rates of assaults. CONCLUSIONS: Although work in health care is associated with high rates of assaults, closer scrutiny suggests specific possible intervention strategies. PMID- 15534502 TI - Respiratory disease in aggregate quarry workers related to risk factors and Pi phenotype. AB - OBJECTIVE: To evaluate the respiratory health and its relationship with exposure to silica, smoking, alpha 1-antitrypsin, and Pi phenotype in aggregate quarries workers. METHODS: In a prevalence study, 378 subjects were studied, x-ray opacities and pulmonary function was analyzed in relation to cumulative dust exposure, smoking, alpha1- antitrypsin, and Pi phenotype. RESULTS: The adjusted odds ratio (OR) of round opacities (> or =0/1) was significantly related to smoking (P <0.001). The predicted forced expiratory volume in 1 second (FEV1) percentage had a negative relationship with the product dust. pack-years (P = 0.005). The OR of FEV1 <90% was 3.03 for smokers exposed to dust versus nonexposed nonsmokers (P = 0.049). The criteria for dust-tobacco interaction are satisfied. A subgroup with the highest level of alpha1-antitrypsin presented more opacities and worse pulmonary function. CONCLUSIONS: Radiologic alterations are found in relation to tobacco. The FEV1 has a negative relation with the product dust-tobacco. No alterations related to Pi phenotype are found. A subgroup with the highest alpha1-antitrypsin levels had more radiologic alterations and worse function. PMID- 15534504 TI - Analysis of text from injury reports improves understanding of construction falls. AB - OBJECTIVE: We combined payroll data, coded workers' compensation (WC) data, and text descriptions of injuries from the construction of Denver International Airport to create a more comprehensive picture of falls from height (FFH) than is typically available from WC data. Text descriptions were coded to identify circumstances surrounding falls. Slips/trips preceded one third of FFH, often involving motor vehicles or heavy equipment. Another third involved movement or collapse of work surfaces, usually ladders or scaffolds. CONCLUSIONS: The significant contribution of motor vehicles and heavy equipment to FFH, particularly those preceded by slips/trips, was not apparent from coded data. Heavy equipment engineering modifications are called for and workers in street/roadway construction/site development need fall protection training. Text analyses allow exploration of factors not identified at the time of data collection and better understanding of the context in which injuries occur. PMID- 15534505 TI - Prevalence of respiratory symptoms in children and air quality by village in rural Indonesia. AB - AIM: This study compared prevalence of respiratory symptoms in three Indonesian villages and related this to air quality. METHODS: We interviewed caregivers of 382 children, using the International Study of Asthma and Allergies in Childhood (ISAAC) questionnaire, and monitored air quality during the survey period. RESULTS: Respiratory symptom prevalence was highest in Kerinci (40.5%), followed by SP7 (33.3%) and Pelalawan (19.8%). Compared with Pelalawan, adjusted odds ratios were 3.17 (95% confidence interval, 1.43-7.07) for Kerinci, and 2.03 (1.04 3.96) for SP7. Ambient air quality levels were highest in Kerinci for PM10 and hydrocarbon (means: 102.9 microg/m3, 10.5 microg/m3), followed by SP7 (73.7 microg/m3, 6.3 microg/m3) and Pelalawan (26.1 microg/m3, 4.7 microg/m3). CONCLUSIONS: The higher prevalence of respiratory symptoms in Kerinci and SP7 could be the result of higher PM10 and hydrocarbon levels in these locations. PMID- 15534506 TI - Biologic markers of oxidative stress and nephrotoxicity as studied in biomonitoring of adverse effects of occupational exposure to lead and cadmium. AB - OBJECTIVES: In this work, we studied impregnation levels of workers occupationally exposed to lead (Pb) and cadmium (Cd), usefulness of early urinary markers of nephrotoxicity, and occurrence of oxidative stress as the underlying mechanism involved in Pb- or Cd-induced adverse effects. Thirty-five men were recruited from a nonferrous metal smelter. Pb and Cd in blood (B-Pb, B-Cd) and urine (U-Pb, U-Cd) were measured. Relations between oxidative stress markers (malondialdehyde, superoxide dismutase, glutathione peroxidase, selenium, glutathione reductase, glutathione status, 8-hydroxy-2'-deoxyguanosine) and exposure levels, on the one hand, and early urinary markers (alpha-1 microprotein, beta-2-microglobulin, retinol binding protein, alpha and pi glutathione S-transferases) and exposure levels, on the other hand, were evaluated. RESULTS: Mean exposure levels were moderate (B-Pb = 395.71 microg Pb/L; U-Pb = 95.19 microg Pb/g creatinine; B-Cd = 5.83 microg Cd/L; U-Cd = 4.67 microg Cd/g creatinine). Changes in malondialdehyde, glutathione status, 8 hydroxy-2'-deoxyguanosine, and alpha-glutathione S-transferases were closely correlated with exposure levels and did not depend on tobacco consumption. We showed that these workers showed moderate Pb and Cd exposure levels. CONCLUSIONS: Taken together, the data suggests the use of alpha-glutathione S-transferases excretion in urine as a hallmark of early changes in the proximal tubular integrity that could later lead to clinical disease if exposure is not reduced. PMID- 15534513 TI - Replacement of the severely damaged arthritic knee by the ICLH (Freeman-Swanson) arthroplasty. 1977. PMID- 15534514 TI - Patella resurfacing versus nonresurfacing in total knee arthroplasty: results of a randomized controlled clinical trial at a minimum of 10 years' followup. AB - Patellar resurfacing in total knee arthroplasty remains controversial. This study evaluates the results of resurfacing and nonresurfacing the patella in a randomized controlled, clinical trial at a minimum of 10 years followup. One hundred knees (90 patients) with osteoarthritis were enrolled in a prospective randomized clinical trial using a posterior-cruciate-retaining total knee arthroplasty. Patients were randomized to receive resurfacing or retention of the patella. Evaluations were done preoperatively and yearly, up to a minimum of 10 years (range, 10.1-11.5 years) postoperatively. Disease-specific (Knee Society clinical rating score) and functional (stair climbing, flexion/extension torques, patellar examination) outcomes were measured. Patient satisfaction, anterior knee pain, and patellofemoral questionnaires were completed. Intraoperative grading of the articular cartilage was done. No patients were lost to followup; 45 patients remained alive. Nine revisions (in nine of 90 knees; 10%) were done in seven patients in the nonresurfaced group (15% of knees) and in two patients in the resurfaced group (5% of knees). No significant difference was found between the groups regarding revision rates, Knee Society clinical rating scores, and functional, patient satisfaction, anterior knee pain, patellofemoral, and radiographic outcomes. Intraoperative cartilage quality was not a predictor of outcome. This study currently is the longest followup of a randomized controlled, clinical trial that examines patellar resurfacing in total knee arthroplasty. The results showed no significant difference between the groups for all outcome measures at a minimum of 10 years of followup. PMID- 15534515 TI - Tibial component failure mechanisms in total knee arthroplasty. AB - The purpose of this study was to examine the failure mechanisms and factors associated with failure of a nonmodular metal backed cemented tibial component. Out of 3152 total knee replacements done for osteoarthritis, 41 tibial components had been revised (1.3%). Four distinct failure mechanisms were identified: 20 knees were revised for medial bone collapse, 13 for ligamentous imbalance, 6 for progressive radiolucencies, and 2 for pain. Factors associated with medial bone collapse were varus tibial component alignment more than 3.0 degrees , Body Mass Index higher than 33.7, and overall postoperative varus limb alignment. Ligamentous imbalance was more prevalent in knees with preoperative valgus deformity. There were no knees revised for tibial component polyethylene wear or osteolysis. We conclude that the dominant failure mechanisms for this component design are related to preoperative deformity, technical factors of component alignment, overall limb alignment, and ligamentous imbalance. PMID- 15534516 TI - Mid-term to long-term followup of two-stage reimplantation for infected total knee arthroplasty. AB - Between January 1989 and December 1994, 94 patients (96 knees) had a two-stage reimplantation for treatment of an infected total knee arthroplasty. All patients were treated with an interval antibiotic-loaded static cement spacer and had antibiotic-loaded bone cement for prosthesis fixation at the time of reimplantation. The purpose of this study was to assess the long-term risk of reinfection and the mechanical durability of these reimplantation arthroplasties. Patients were followed up for a median of 7.2 years (range, 2.5-13.2 years). At final followup, 15 knees (16%) had required reoperation. Nine knees (9%) had component removal for reinfection and six knees (6%) were revised for aseptic loosening. The median time to reoperation for reinfection was 1 year (range, 0.1 9.8 years). The risk of recurrent infection was not correlated with the type of organism, patient demographics, or method of prosthesis fixation at reimplantation. The survivorship free of implant removal for any reason was 90% (confidence intervals, 83.9-96.4%) at 5 years and 77.3% (confidence intervals, 65.5-89.6%) at 10 years. The survivorship free of implant removal for reinfection was 93.5% (confidence intervals, 88.5-98.7%) at 5 years and 85% (confidence intervals, 73.8-96.3%) at 10 years. Survival free of revision for mechanical failure (aseptic loosening or radiographic loosening) was 96.2% (confidence intervals, 92-100%) at 5 years and 91% (confidence intervals, 80.8-98.3%) at 10 years. These results suggest that the high likelihood of early success after two stage reimplantation of an infected TKA is well maintained throughout long-term followup, with a modest rate of late recurrent infection or mechanical implant failure. PMID- 15534517 TI - Factors influencing wear and osteolysis in press-fit condylar modular total knee replacements. AB - The purpose of this study was to determine the factors influencing wear and osteolysis in patients who have had total knee arthroplasty with the Press-Fit Condylar modular system. Two-thousand ninety-one primary total knee replacements in 1737 patients were done using the Press-Fit Condylar system at three centers. Radiographic and manufacturing data were obtained for 2016 of the 2091 implants (96.4%). For the 1287 of 2016 knees (64%) with more than 5 years of followup, the prevalence of wear-related failure was 8.3%. The 13-year survivorship for all patients was 82.6%. Cox hazards analysis revealed five variables that were correlated with wear-related failure: patient age, patient gender, polyethylene sheet vendor, polyethylene finishing method, and polyethylene shelf age. We were unable to identify one factor as the defining reason for these wear-related failures. The multiple changes in manufacturing methods during the life of this implant may have precluded such a determination. These results may be specific to inserts sterilized in air with gamma irradiation and should not be generalized to current manufacturing techniques. This study emphasizes the potential deleterious effects that small changes in the manufacturing process may have on the outcome of a prosthesis with an initially favorable survivorship. PMID- 15534518 TI - Minimally invasive knee arthroplasty. AB - Interest in minimal-incision surgery among physicians and patients has led to the investigation of whether minimal-incision concepts have a role in knee arthroplasty. Successful outcomes in knee arthroplasty have been traditionally measured by long-term implant performance and low revision rates, with less emphasis on the size of the incision or the length of time to complete recovery. There are two evolving lines of development in minimal-incision knee arthroplasty: the small-incision approach and the new technology approach. The small incision approach seeks to minimize the length of the incision required to implant standard total knee components by altering the approach and instrumentation. The new technology approach seeks to develop lower profile implants and computer-assisted techniques to fundamentally change the way knee arthroplasty is done. The burden of proof as to whether these new approaches will stand the test of time remains with the investigators. PMID- 15534519 TI - MIS unicondylar knee arthroplasty: surgical approach and early results. AB - Unicondylar arthroplasty of the knee has seen a resurgence of interest in the United States. The principles of unicondylar arthroplasty of the knee are different from those for total knee arthroplasty, allowing replacement of only the affected joint compartment with less bone loss. Minimally invasive surgery allows for less soft tissue dissection with the potential for less morbidity. The key question is: will the changes associated with the minimally invasive surgery procedure improve the clinical results of the standard unicondylar arthroplasty of the knee or will the changes make the procedure too difficult and lead to an increasing failure rate? This study reviews the surgical technique and presents the 2 to 4 year results of the minimally invasive unicondylar arthroplasty of the knee 47 knees in 41 patients. The average range of motion increased from 121 degrees -132 degrees . The Knee Society pain score improved from 45-80 and the function score improved from 47-78. Only one knee has been revised. With proper patient selection, minimally invasive unicondylar arthroplasty of the knee allows for results that are at least equal to those of the standard open procedure at 2 to 4 years after the surgery. PMID- 15534520 TI - Surgical approaches in mini-incision total knee arthroplasty. AB - Mini-incision total knee arthroplasty can be accomplished through versions of exposures used in standard total knee arthroplasty. Modifications of the medial parapatellar, subvastus, and midvastus approaches are presented, and potential advantages and disadvantages of each approach are reviewed. When making the transition to smaller incisions and arthrotomies, the medial parapatellar seems to be the most versatile. The ability to do a total knee arthroplasty through a mini incision, and ultimately the success of the procedure, will depend on appropriate patient selection. PMID- 15534521 TI - Minimally invasive total knee replacement through a mini midvastus approach: a comparative study. AB - Between September 2001 and September 2002, forty consecutive minimally invasive total knee replacements were done. A modified midvastus approach was used and the patella was subluxed, but not everted. We compared the results of this group with an age-matched and sex-matched cohort of total knee replacements done between June 2000 and September 2001 with a standard technique. A posterior-stabilized knee (Genesis II) was used in both groups. Patients achieved motion considerably faster in the minimally invasive total knee replacement group. Mean flexion for minimally invasive total knee replacement at 6 and 12 weeks was 114 degrees (range, 90-132 degrees ) and 122 degrees (range, 103-135 degrees ) respectively, compared with 95 degrees (range, 65-125 degrees ) and 110 degrees (range, 80-125 degrees ) for the control group. Improved range of motion was also seen at one year postoperatively. The average range of motion at one year postoperatively in the minimally invasive total knee replacement was 125 degrees (range, 110-135 degrees ) compared with 116 degrees (range, 95-130 degrees ) in the Control Group. Postoperative Knee Society scores were also higher in the minimally invasive total knee replacement group. There was no difference in xray alignment. There were no infections, extensor mechanism or neurovascular complications. The mini midvastus approach without patella eversion combined with a small incision was associated with a more rapid functional recovery and improved range of motion in total knee replacement without compromising implant positioning. PMID- 15534522 TI - Minimally invasive total knee replacement through a mini-midvastus incision: an outcome study. AB - Total knee replacement traditionally has been done through an anterior incision approximately 18 cm long, using a capsular incision that separates the interval between the rectus femoris and vastus medialis musculature. Although giving excellent exposure, this incision also disrupts the suprapatellar pouch and may lead to adhesions and difficulty with rapidly regaining flexion. It is hypothesized that, by using a more minimally invasive incision, there will be a more rapid return of flexion and the patient will require fewer narcotic medications postoperatively. This retrospective review compared 32 total knee replacements done through a minimally invasive mini-midvastus approach with 26 total knee replacements done through the standard medial parapatellar approach. Preoperative Knee Society scores and postoperative functional outcomes were compared. Postoperative flexion was measured daily during hospitalization and at a 6-week and 3-month followup. Pain was assessed by a visual analog scale and the amount of pain medication used during hospitalization. Implant position was measured. The MIS group had an average skin incision length of 12.8 cm. Passive flexion on a daily basis was significantly higher in the MIS group compared with the standard group. At 6 weeks postoperatively, the change in Knee Score was statistically higher in the MIS group and the average visual analog pain score and the total amount of pain medication was lower. The radiographic alignment and position of all the components was normal in all patients in both groups. The limited disruption of the extensor mechanism results in more rapid restoration of the quadriceps muscle control. PMID- 15534523 TI - Debate: bilateral simultaneous total knee arthroplasty. AB - Controversy persists regarding the value of doing bilateral simultaneous total knee arthroplasty. Advocates of this procedure focus on economic costs, patient satisfaction, and quicker return to function as compared with bilateral staged total knee arthroplasty. Those in opposition focus on increased complication rates that question the overall safety of this operative procedure. The purpose of this discussion is to review the orthopaedic literature, concentrating on the reported advantages and disadvantages of bilateral simultaneous total knee arthroplasty in hopes of providing the surgeon with information valuable in determining the safety and efficacy of this procedure. PMID- 15534524 TI - Debate: simultaneous bilateral knee replacements: the outcomes justify its use. AB - The purpose of this paper is to assess the morbidity, mortality, and clinical outcome of simultaneous bilateral total knee arthroplasty. We reviewed 4100 simultaneous bilateral total knee replacements. The knees were subjected to two Kaplan-Meier survival analyses, with failure equal to revision for aseptic loosening and failure equal to patient death. Complications and Knee Society scores were considered. The average Knee Society knee score was 90 points 3 years postoperatively and 87 points 10 years postoperatively. The complication rates were as follows: deep infection (0.8%), superficial infection (0.3%), cardiac (6 arrhythmia, 5 congestive heart failure, 1 cardiac insufficiency, 3 complete heart block, 2 myocardial infarction and cardiac arrest, and 14 myocardial infarction only) (1.5%), intestinal ileus (0.5%), gastrointestinal ulcer (0.4%), thrombophlebitis (0.9%), cerebrovascular accident (0.3%), and urinary (1 BPH obstruction, 4 renal failure, 2 transurethral resection of the prostate, 16 urinary tract infection, and 2 urinary retention/incontinence) (1.2%). The 10 year prosthesis survival probability was 98.3%. The 10-year patient survival probability was 78.6%. Twenty-five (1.2%) patients died within the first postoperative year. The patients who died within 1 year postoperatively were older than the rest of the group. Higher age and male gender were factors related to increased mortality. The complication rates and clinical outcomes were similar to unilateral total knee arthroplasty. With regard to death early in the postoperative course, simultaneous bilateral total knee arthroplasty may pose a greater risk to the patient than a unilateral procedure. However, the early deaths may be related to older age at the time of surgery. PMID- 15534525 TI - Bilateral total knee replacement using the same anesthetic is not justified by assessment of the risks. AB - The question of whether simultaneous bilateral knee replacement is associated with an increased risk of perioperative morbidity and mortality remains controversial. There are no true prospective randomized studies that address this question. There is general agreement that the risk is higher for older patients and for those with considerable medical comorbidities, particularly those with a history of cardiac disease. There is no general agreement on the appropriate threshold for age or comorbidities in order to minimize risk. Based on available retrospective data, there is evidence to suggest that there may be some patient selection bias favoring patients with simultaneous bilateral knee replacements; despite this bias, the perioperative risks are higher for these healthier patients. The highest risk for the patient having simultaneous bilateral knee replacement seems to be that of early postoperative mortality followed by the risk of a cardiac event. The risks of thromboembolic, gastrointestinal, and neurologic dysfunction also are increased. These patients also have a higher likelihood of requiring homologous blood transfusions and of requiring posthospitalization transfer to a rehabilitation center. Because of the absence of established patient selection criteria, thresholds for assessment of the increased risks, the patient having simultaneous bilateral replacement faces an increased risk of perioperative mortality and morbidity on a random basis. Patients should be aware of this information when deciding whether to proceed with simultaneous bilateral knee replacement. PMID- 15534526 TI - The progression of patellofemoral arthrosis after medial unicompartmental replacement: results at 11 to 15 years. AB - This study reports the 11-year to 15-year results of unicompartmental knee arthroplasty with an emphasis on failure mechanisms and progression of patellofemoral arthrosis. In a prospective study of 513 consecutive potential knee replacement candidates, 59 patients (12%) had medial unicompartmental arthroplasty of the knee. All 59 patients had isolated unicompartmental disease without clinical symptoms or radiographic evidence of patellofemoral arthritis. No patient was lost to followup. The average followup was 13 years (range, 11-15 years). The mean preoperative Hospital for Special Surgery knee score of 55 points (range, 30-79 points) improved to a mean of 90 points (range, 60-100 points) at final followup. Patellofemoral symptoms were present in 1.6% of patients at 10 years; this increased markedly to 10% of patients at 15 years (p < 0.01). Four patients (10%) had moderate or severe patellofemoral symptoms at final followup; two were revised to a primary total knee replacement at 7 and 11 years for progressive patellofemoral degeneration. No component was radiographically loose and no osteolysis was seen. The Kaplan-Meier survival with loosening or revision for any reason was 98.0% +/- 2.0% at 10 years and 95.7% +/- 4.3% at 15 years. At up to 15 years, unicompartmental knee arthroplasty yielded good clinical results; however, progressive patellofemoral arthritis was the primary mode of failure. PMID- 15534527 TI - Why are total knee replacements revised?: analysis of early revision in a community knee implant registry. AB - Since 1991, 5760 knee arthroplasty procedures done by 53 surgeons have been registered in a community joint implant registry and were reviewed regarding initial revision done within the healthcare system. The 168 revisions done represented 2.9% of the knee arthroplasties between September 1991 and December 2002. Survival was defined as the absence of revision surgery. Death was considered a censored event. Cumulative survival rates for the different total knee arthroplasty configurations were: cemented total knee arthroplasty with all polyethylene tibia, 99.2%; cemented total knee arthroplasty with metal-backed tibia, 96.3%; hybrid total knee arthroplasty, 89.3%; and unicondylar knee arthroplasty, 87.2%. Cemented total knee arthroplasty with metal-backed tibia had better survival than hybrid total knee arthroplasty, ingrowth total knee arthroplasty, and unicondylar knee arthroplasty. Cemented total knee arthroplasty with a metal-backed tibia did not have better survival than cemented total knee arthroplasty with an all-polyethylene tibia. Gender was not related to survival. Age was related to survival, with older patients' knees surviving longer. Aseptic loosening or wear was the cause of revision in 40.8% of patients having total knee arthroplasty and 46.6% of patients having unicondylar knee arthroplasty, whereas progression of arthritis necessitated unicondylar knee revision in 51.2% of patients having that procedure. This study presents further evidence of the value of and ongoing need for total joint registries. Cemented total knee arthroplasty with all-polyethylene tibia and with metal-backed tibia showed more than 95% 10-year cumulative survival. Hybrid total knee arthroplasty, ingrowth total knee arthroplasty, and unicondylar knee arthroplasties did not show such good results. PMID- 15534529 TI - Knee simulator wear of polyethylene tibias articulating against explanted rough femoral components. AB - Highly cross-linked and melted polyethylene tibial inserts have recently been introduced for clinical use to reduce fatigue damage and adhesive wear in tibial inserts. Other authors have studied the effect of counterface roughness on the wear behavior of polyethylene tibial inserts in knee simulators using femoral components that were roughened artificially. They reported a higher wear rate with highly cross-linked polyethylene than with unirradiated polyethylene tibial inserts. Artificial roughening of femoral components may not be clinically relevant. To evaluate this concern, we studied the wear behavior of highly cross linked and conventional polyethylene tibial inserts articulating in vitro against surgically retrieved femoral components that had become roughened in vivo. The wear rate of the highly cross-linked polyethylene (5.9 and 6.8 mg/1 million cycles with 100 and 50% serum) was 80% lower than the wear rate of the conventional polyethylene (33.5 and 32.2 mg/1 million cycles with 100 and 50% serum) tibial inserts after 2 million cycles of simulated gait. This study suggests that during in vivo use, scratches that are generated on the femoral components are likely to produce a higher wear rate with both cross-linked and conventional polyethylene than a smooth femoral component, but that this wear rate is likely to be higher with conventional polyethylene than with highly cross linked polyethylene tibial inserts. PMID- 15534530 TI - Wear, debris, and biologic activity of cross-linked polyethylene in the knee: benefits and potential concerns. AB - Cross-linked polyethylene currently is being introduced in knee prostheses. The wear rates, wear debris, and biologic reactivity of non cross-linked, moderately cross-linked, and highly cross-linked polyethylene have been compared in multidirectional wear tests and knee simulators. Multidirectional pin-on-plate wear studies of noncross-linked, moderately cross-linked (5 Mrad), and highly cross-linked (10 Mrad) polyethylene showed a 75% reduction in wear with the highly cross-linked material under kinematics found in the hip, but only a 33% reduction under wear in kinematics representative of the knee. In knee simulator studies, with the fixed-bearing press-fit, condylar Sigma cruciate-retaining knee under high kinematic input conditions, the wear of 5 Mrad moderately cross-linked polyethylene was 13 +/- 4 mm per 1 million cycles, which was lower (p < 0.05) than the wear of clinically used, gamma vacuum foil GUR 1020 polyethylene (23 +/- 6 mm/1 million cycles). For the low-contact stress mobile-bearing knee, the wear of moderately cross-linked polyethylene was 2 +/- 1 mm per 1 million cycles, which was lower (p < 0.05) than GVF GUR 1020 polyethylene (5 +/- 2 mm/1 million cycles). The wear debris isolated from the fixed-bearing knees showed the moderately cross-linked material had a larger percentage volume of particles smaller than 1 mum in size, compared with GVF GUR 1020 polyethylene. Direct cell culture studies of wear debris generated in sterile wear simulators using multidirectional motion showed a increase (p < 0.05) in tumor necrosis factor alpha levels and reactivity for GUR 1050 cross-linked polyethylene debris compared with an equivalent volume of noncross-linked GUR 1050 polyethylene. The use of cross-linked polyethylene in the knee reduces the volumetric wear rate. However, the clinical significance of reduced fracture toughness, elevated wear in abrasive conditions, and the elevated tumor necrosis factor-alpha release from smaller more reactive particles warrant further investigation. PMID- 15534531 TI - Oxidized zirconium femoral components reduce polyethylene wear in a knee wear simulator. AB - Polyethylene wear remains a major problem that can jeopardize the long-term durability of prostheses used in total knee arthroplasty. Oxidized zirconium is a material that combines the strength of a metal with the wear properties of a ceramic. This study evaluated the wear rates of polyethylene inserts against oxidized zirconium femoral components. Three oxidized zirconium femoral components and three Co-Cr femoral components of identical geometry used in total knee arthroplasty were articulated against standard tibial components with modular tibial inserts made of noncross-linked ultra-high molecular weight polyethylene. Gravimetric and volumetric polyethylene wear rates were calculated after 5 million gait cycles on an AMTI knee wear simulator. Oxidized zirconium reduced polyethylene wear by 42% compared with Co-Cr alloy. This study shows that oxidized zirconium can reduce polyethylene wear substantially when used for fixed bearing total knee arthroplasty. PMID- 15534532 TI - Soft tissue and intra-articular injection of bupivacaine, epinephrine, and morphine has a beneficial effect after total knee arthroplasty. AB - The purpose of this study was to determine if an intraoperative intraarticular and soft-tissue injection of local anaesthetic, epinephrine, and morphine has a beneficial effect for total knee arthroplasty. A control group of 138 patients (181 knees) received no intraoperative injection. The study group of 171 patients (197 knees) received intraoperative injection of 0.25% bupivacaine with epinephrine and morphine with 2/3 injected into the soft tissues and 1/3 injected into the joint. Patients having bilateral simultaneous procedures received a divided dose. The pain treatment protocol otherwise was identical. Pain, sedation, rescue narcotic usage, narcotic reversal and blood loss were examined. Pain levels during the immediate postoperative period, blood loss, and bleeding indices were reduced with injection. Considerably more control patients required rescue doses of narcotics. Preemptive analgesia with soft tissue and intra articular injection of long-acting local anesthetic with epinephrine and morphine provides better pain control in the immediate postoperative period, decreases blood loss, and decreases the need for rescue narcotics and reversal agents. This simple, inexpensive method provides an effective adjunct to a multimodal approach in improving the postoperative course of primary total knee arthroplasty. PMID- 15534533 TI - Image-based computer-assisted total knee arthroplasty leads to lower variability in coronal alignment. AB - In a prospective randomized controlled trial, one group of 50 patients had total knee arthroplasty using conventional instruments; another group of 50 patients had total knee arthroplasty using a fluoroscopy-based computer navigation system. The variability of postoperative alignment of the lower limb in the coronal and saggital plane, early clinical outcome scores, and morbidity of the procedure were compared. In addition, the image-based computer navigation system was used to assess the following presumptions: the accuracy of the calculation of the kinematic center of rotation of the hip and the reliability of full-leg standing xrays in determining overall coronal alignment of the lower limb. Variability in the coronal plane was significantly reduced in the computer-assisted surgery group compared with the conventional group (p < 0.0001). Early clinical outcome and complication rates were similar for both groups. The correlation between full leg standing xrays and the computer navigation system for determining the mechanical alignment of the lower limb in the coronal plane was high (r = 0.987). The maximum deviation between the calculated kinematic center of rotation of the hip and the fluoroscopically determined anatomic center of the hip was 5 mm (mean, 1.6 mm). An image based computer-assisted system can be safely used and leads to a consistent and perfect postoperative alignment in the coronal plane. PMID- 15534534 TI - Osteogenic protein-1 in knee arthritis and arthroplasty. AB - The use of graft materials to restore bone stock and promote healing and implant stabilization is a crucial part of total knee arthroplasty, especially in revision surgery. Recent research has centered on the use of osteoinductive materials to promote bone formation. Osteogenic proteins are members of a superfamily of proteins called transforming growth factor-beta that, either alone or in combination with other regulatory molecules, induce new bone formation. The cloning and genetic expression of recombinant human osteogenic proteins has led to production of quantities sufficient for their clinical use. Recombinant human osteogenic protein-1 has been combined with bone-derived Type I collagen for delivery to an implant site. Preclinical studies have shown that the osteoinductive capacity of autograft and allograft bone and bone graft substitute materials can be notably improved with the addition of osteogenic protein-1. The use of this protein consistently improved the amount and rate of new bone formation compared with graft alone, resulting in earlier graft incorporation and consolidation. In addition, because osteogenic proteins are chondrogenic, they also may have a role in the treatment of cartilage injury and degeneration. Osteogenic protein-1 has been shown to induce hyalinelike cartilage repair of full thickness osteochondral defects in animal models with no degradation of the tissue with time. Although no detailed clinical studies in knee surgery have been reported with the use of osteogenic protein-1, in anecdotal cases its use alone and with bone graft materials indicate results consistent with those obtained in preclinical studies. PMID- 15534535 TI - Long-term followup of posterior-cruciate-retaining TKR in patients with rheumatoid arthritis. AB - A consecutive series of 220 primary posterior-cruciate ligament-retaining total knee replacements were done in 148 patients with rheumatoid arthritis. From this group, 212 total knee replacements (141 patients) were followed up for an average of 10 years. Average knee scores at 5, 10, 15, and 20 years after operation improved to 86, 83, 88, and 89, respectively, and average function scores improved to 70, 72, 64, and 88, respectively. Five knees (2.4%) were revised for deep infection. Posterior instability, recurvaum, or mediolateral instability, combined or otherwise, occurred in 15% (32 knees; in 31 patients). Three tibial components (1.4%) were revised: one for suspected aseptic loosening and two for instability. Excluding infections and failed metal-backed patellas, Kaplan-Meier survival rates were 99.5%, 97.9%, and 96.5%, respectively. Favorable long-term results may be achieved with posterior-cruciate ligament-retaining total knee replacements in patients with rheumatoid arthritis. Concern remains, however, about rotational instability in certain cases associated with preoperative genu valgum and ipsilateral planovalgus deformity. PMID- 15534536 TI - A comparison of highly instrumented and minimally instrumented unicompartmental knee prostheses. AB - In this study, two specific implants are compared: the Oxford prosthesis, which uses a highly instrumented technique and was placed through a traditional arthrotomy; and the Repicci prosthesis, which uses a minimally instrumented technique and was placed with a minimally invasive surgical approach. The study looked specifically at limb alignment, because achieving appropriate alignment is correlated with the best long-term implant function and achieving alignment is a function of the implant instrumentation. The rationale of more instrumentation for a surgical technique is to add precision and reproducibility; less instrumentation requires less surgical exposure and therefore is more minimally invasive. The senior author presents his long-term experience with the Oxford implant (55 implants; average followup 10.4 years) and short-term experience with the Repicci implant (164 implants; average followup 1.3 years for the first 30 implants.). With available followup, the clinical results using both implants have been favorable. The average alignment of the Oxford prosthesis was 5.6 degrees valgus and the average alignment of the Repicci prosthesis was 4.5 degrees valgus. Side-to-side long-term comparison is not yet available, although the literature would suggest that both implants can have satisfactory long-term results. Specific implant selection probably is not as important as precise patient selection. PMID- 15534537 TI - Patellofemoral arthroplasty: pros, cons, and design considerations. AB - Patellofemoral arthroplasty is a worthy alternative to total knee arthroplasty or patellectomy in patients with arthritis localized to the anterior compartment of the knee, particularly when there is no considerable patellar malalignment or maltracking. The results can be optimized by accurately aligning the prosthesis and balancing the soft tissues to enhance patellar tracking. However, some designs are particularly vulnerable to patellofemoral complications. Postoperative patellofemoral dysfunction should be reduced by using a trochlear component that engages the patella within the trochlear groove and articulates with the patella completely in extension, but which is relatively unconstrained in extension and has a sagittal radius of curvature that fits well with the native distal femur. The incidence of patellofemoral complications was reduced in my series from 17% with a first generation implant to 4% with a second generation implant. Evolving designs likely will eliminate many of the complications of early generation implants, leaving tibiofemoral degeneration the major source of failure of patellofemoral arthroplasties. PMID- 15534538 TI - The consequences of not resurfacing the patella. AB - The decision of whether to resurface the patella during total knee arthroplasty remains controversial. In this study, a meta-analysis of national joint replacement registry data, bilateral total knee replacement studies, selective resurfacing reports, and randomized clinical trials was done Although the evidence seems to support patellar resurfacing, this issue remains inconclusive because of problems generalizing from one implant to another and the short-term nature of available studies. Based on existing data, patellar resurfacing seems reasonable in most total knee replacements. Not resurfacing the patella might be considered in selected younger patients (<60 years) with mild or no patellar arthritis, a well-tracking extensor mechanism, and particularly if a patella friendly femoral component is used. PMID- 15534539 TI - Is an intact anterior cruciate ligament needed in order to have a well functioning unicondylar knee replacement? AB - Controversy exists about whether a functional anterior cruciate ligament is necessary to achieve success with unicompartmental knee arthroplasty. When the anterior cruciate ligament was deficient, higher failure rates were reported with mobile-bearing implants and with the Lotus implant, a relatively flat, fixed bearing component. Most failures were secondary to wear. In contrast, the absence of an anterior cruciate ligament did not lead to failure with the St. Georg and Marmor implants. Theoretically, an absent anterior cruciate ligament would increase the sliding motion that caused accelerated polyethylene wear in laboratory studies. Hypothetically, such motion could lead to accelerated wear of unicompartmental arthroplasty in an ACL deficient knee. Currently, unicompartmental knee arthroplasties should not be done in patients with symptoms of anterior cruciate ligament instability and should judiciously be done in older patients without a functional anterior cruciate ligament but with no symptoms of instability. PMID- 15534540 TI - A high flexion total knee arthroplasty design replicates healthy knee motion. AB - Deep flexion affects both femorotibial contact pattern and the patellofemoral articulation. The purpose of this study was to compare the patellofemoral motion of nonimplanted and implanted knees and to analyze femorotibial kinematics after total knee replacement designed for deep flexion. Three-dimensional patellofemoral kinematics were evaluated during a deep knee bend using fluoroscopy for five control patients with a healthy knee, five patients with an anterior-cruciate-ligament-deficient knee, and 20 patients who had a high flexion total knee arthroplasty. Less translation of patellofemoral contact position was seen in patients who had knee replacements than in patients with healthy knees, but the average motion and the patella tilt angles were similar to the healthy knees. On average, patients who had a total knee arthroplasty had 4.9 degrees normal axial rotation, and all patients had at least -4.4 mm of posterior femoral rollback. The average weightbearing range of motion of the patients in the total knee arthroplasty group was 125 degrees . In this study, patients implanted with a high-flexion knee replacement design had kinematic patterns that were similar to the healthy knee. It can be hypothesized that forces acting on the patella were not substantially increased for patients who had a total knee arthroplasty compared with the control patients. PMID- 15534541 TI - A multicenter analysis of axial femorotibial rotation after total knee arthroplasty. AB - A multicenter analysis was done to determine in vivo femorotibial axial rotation magnitudes and patterns in 1,027 knees (normal knees, nonimplanted ACL-deficient knees, and multiple designs of total knee arthroplasty). All knees were analyzed using fluoroscopy and a three-dimensional computer model-fitting technique during a deep knee bend and/or gait. Normal knees showed 16.5 degrees and 5.7 degrees of internal tibial rotation during a deep knee bend and gait, respectively. Rotation magnitudes and the percent having normal axial rotation patterns decreased in all total knee arthroplasty groups during a deep knee bend. During gait, all knee arthroplasty groups had similar rotational patterns (limited magnitudes). Average axial rotational magnitudes in gait and a deep knee bend were similar among major implant categories (ie, fixed-bearing versus mobile-bearing, etc). Average values in normal knees and ACL-retaining total knee arthroplasty patients (16.5 degrees and 8.1 degrees , respectively) were higher than in groups in which the ACL was absent (< 4.0 degrees ). All total knee arthroplasty groups had at least 19% of patients have a reverse axial rotational pattern during a deep knee bend and at least 31% during gait. Normal axial rotation patterns are essential for good patellar tracking, reduction of patellofemoral shear forces, and maximization of knee flexion. PMID- 15534542 TI - Extended medial gastrocnemius rotational flap for treatment of chronic knee extensor mechanism deficiency in patients with and without total knee arthroplasty. AB - Nine patients with chronic extensor mechanism disruption were treated with an extended medial gastrocnemius rotational flap reconstruction of the extensor mechanism. Seven patients previously had total knee arthroplasty and two patients had chronic infection of nonreplaced, native knees. Four patients previously had failed Achilles' tendon allograft reconstruction after total knee arthroplasty and two were complicated by infection. Infected arthroplasty patients had a staged procedure with placement of an antibiotic spacer after debridement and extended medial gastrocnemius rotational flap, followed by total knee arthroplasty replant 8 weeks later. The four infected arthroplasty patients had medical comorbidities that included a patient with HIV and hemophilia, and two with diabetes mellitus. Another patient with rheumatoid arthritis was severely malnourished as a result of dumping syndrome. Of the four patients treated by this two-stage procedure, one died in the early postoperative period from chronic medical issues after the second stage and another patient elected to have above knee amputation after the first stage because of severe reflex sympathetic dystrophy. The final group of seven patients was studied at a mean followup of 21 months (range, 7-31 months), the average extensor lag was 13.5 degrees (range, 0 50 degrees ), and the average range of motion was 2 degrees to 93 degrees . The two patients with nonreplaced, native knees had extensor lags of 30 degrees and 10 degrees . All patients were able to regain sufficient extensor mechanism strength to return to independent ambulation, and all infections resolved after treatment. Two patients were able to ascend stairs foot over foot without support. In addition to the patient who had amputation, the other complication involved a wound breakdown that required a free flap at 13 months in a patient who had a failed Achilles' tendon allograft reconstruction after takedown of a knee fusion. Medial gastrocnemius flap reconstruction can provide successful salvage of a failed extensor mechanism allograft or an alternative to allograft reconstruction in patients with poor soft tissue coverage, previous infection, or a compromised immune system. PMID- 15534543 TI - Backside wear of Miller-Galante I and Insall-Burstein II tibial inserts. AB - The objective of this study was to determine if there was a difference in the amount, type, and location of backside wear in the Miller-Galante I and Insall Burstein II PE tibial inserts. A secondary objective was to determine if backside wear damage in these two designs was a function of clinical factors (patient height, weight, gender, age, and length of time of implantation), shelf life of the PE tibial insert, and tibial component thickness. Backside wear damage was assessed on 24 Miller-Galante I and 11 Insall-Burstein II tibial inserts (implantation time, 0.5-12.4 years). For both groups combined, implantation time was positively correlated to wear damage and to PE peg height into screw holes. The Miller-Galante I group had significantly larger PE pegs than the Insall Burstein II group. The Miller-Galante I group had significantly more burnishing and larger PE pegs posteriorly than anteriorly. There was no correlation between insert shelf life before initial surgery and backside wear. The thinner the component, the larger the total damage scores in the Miller-Galante I group. This study supports the hypothesis that backside wear of PE tibial inserts may be influenced by design and component thickness and by clinical factors. PMID- 15534544 TI - In vivo polyethylene bearing mobility is maintained in posterior stabilized total knee arthroplasty. AB - In vivo knee kinematics, including polyethylene bearing mobility, were determined in a group of nine patients implanted with a posterior stabilized, mobile-bearing total knee arthroplasty. Each patient, while under fluoroscopic surveillance, did a weightbearing deep knee bend and was analyzed using a 3-D computer model fitting technique. Patients were evaluated at three and 15 months postoperatively. All nine patients had polyethylene bearing rotation relative to the tibial tray at both times, with the maximum amount of polyethylene bearing rotation at any flexion interval averaging 8.5 (range, 5.2-15.5) and 9.8 (range, 4.8-14.2) at 3 and 15 months, respectively. Minimal rotation of the polyethylene bearing relative to the femoral component was observed, averaging only 1.9 and 1.0 of rotation from full extension to maximum knee flexion at three and 15 months, respectively. This study determined that the polyethylene bearing is primarily rotating relative to the tibia rather than the femoral component. Therefore, as the femoral component axially rotates, the polyethylene bearing is rotating a similar magnitude in the same direction. This should result in reduced shear stresses on the superior aspect of the polyethylene bearing, lessening polyethylene wear. PMID- 15534545 TI - The outcome of the cementless tibial component: a minimum 14-year clinical evaluation. AB - The long term survival of the Miller-Galante I cementless total knee arthroplasty was evaluated by studying prospectively for a minimum of 14 years (range, 14-17 years), 124 consecutive cementless total knee arthroplasties using a Miller Galante I prosthesis in 99 patients with a mean age of 62 years who had primary or secondary osteoarthritis. Knee function and roentgenograms were evaluated using the Knee Society criteria. Kaplan-Meier survivorship analysis was conducted. Five patients (five knees) were lost to followup; six patients died with six knees in place. Fifteen knees (15 patients) failed and were revised. Thirteen metal-backed patellas were revised. Eight of these revisions also required exchange of the femoral component, but only one tibial tray was revised. One knee (one patient) was revised for an infection. Of the 98 knees not revised (73 patients) observed throughout this study, the average preoperative knee score was 31 (range, 0-47); postoperatively, the average knee score was 91 (range, 72 100). The average function score improved from 28 (range, 10-45) to 84 (range, 50 100). Twenty-four tibial trays (21%) and twenty femoral components (17%) of the 113 knees studied showed osteolysis. This study indicates that osteointegration of cementless tibial components can be successful with screw fixation, although there is a worrisome incidence of tibial and femoral osteolysis. The overall knee survival rate was 87%; however, the tibial component had a survival of 99%. PMID- 15534546 TI - Rotating platform knees did not improve patellar tracking: a prospective, randomized study of 240 primary total knee arthroplasties. AB - Renewed interest in mobile-bearing total knee replacement designs has been generated by the concept of self alignment and the suggestion that those designs can accommodate small mismatches in the rotational position of the tibial and femoral components. Self alignment might improve patellar tracking, decrease the prevalence of lateral retinacular release and postoperative patellar tilt or subluxation, improve knee flexion, and improve patellofemoral function during daily activities such as stair climbing. This prospective randomized study of 240 patients used a single posterior-stabilized femoral component and included three groups of 80 patients: an all-polyethylene group, a modular metal-backed group, and a rotating platform tibia group. The prevalence of lateral retinacular release was 3.8% in each group. The prevalence of patellar tilt was 5% (all polyethylene group), 7% (modular metal-backed group), and 11% (rotating platform group). Preoperative motion was not significantly different and both the 3-month flexion (112 degrees , 110 degrees , and 108 degrees ) and 1-year flexion (116 degrees , 117 degrees , and 115 degrees ) were not significantly different among the all-polyethylene, modular metal-backed, and rotating platform groups, respectively. Preoperative stair climbing scores were not significantly different and both the 3-month (38, 41, and 35 points) and 1-year (44, 46, and 42 points) scores were not significantly different. In this prospective randomized study, the rotating platform knee design did not decrease the prevalence of lateral retinacular release or patellar tilt or subluxation and did not increase knee flexion or improve stair climbing ability at 3 months or at 1 year postoperatively when compared with a posterior-stabilized, fixed-bearing knee. PMID- 15534547 TI - Transfer of the long head of the triceps tendon for irreparable rotator cuff tears. AB - This study presents the clinical and anatomic findings of the use of the long head of the triceps tendon for treatment of massive, irreparable rotator cuff tears. The clinical study included 19 shoulders in 18 patients with a minimum followup of 2 years who were prospectively reviewed. All patients had preoperative and postoperative functional evaluations using the UCLA scoring system. The anatomic study included 20 upper extremities that had been injected with Microfil. All patients in the clinical study were satisfied with their outcome. There was an improvement in the UCLA pain and function scores. The long head of the triceps tendon is a myotendinous vascularized transfer that is a useful reconstructive procedure in patients with massive, irreparable rotator cuff tears that continue to be symptomatic despite conventional attempts at repair. PMID- 15534548 TI - Surgeons rarely discuss sexual activity with patients after THA: a survey of members of the American Association of Hip and Knee Surgeons. AB - A survey of members of the American Association of Hip and Knee Surgeons was done to document surgeons' attitudes and practices regarding discussion of return to sexual activity, timing of return, and safety of various sexual positions for patients after total hip arthroplasty. Surveys were returned by 254 of 821 surgeons (31%). Two hundred thirty-three (80%) surgeons reported they rarely or never discuss sexual activity with their patients who have had hip arthroplasty. Of surgeons who stated they did discuss this topic, 96% spent 5 minutes or less. Most respondents (67%) recommended delaying return to sexual activity until 1-3 months postoperatively. Fifty-one respondents (20%) reported knowledge of patients experiencing dislocation of the total hip replacement during sexual activity. Five specific sexual positions for men and three positions for women were considered acceptable by more than 90% of respondents. Most agreed that sexual activity could be resumed safely 1-3 months postoperatively if acceptable positions were used. PMID- 15534549 TI - The incorporation of morselized bone grafts in cementless acetabular revisions. AB - The treatment of large osteolytic lesions is a challenge during acetabular revisions. Periprosthetic bone loss can compromise the stability of new implants. The purpose of this study was to evaluate the 5- to 10-year clinical and radiographic results of morselized bone grafting for acetabular osteolysis during cementless acetabular revisions. Ninety-nine patients (108 hips) who had an acetabular revision hip arthroplasty for osteolysis were retrospectively reviewed. There were 44 men and 55 women who had a mean age of 66 years. At the index revision, the acetabular defects were debrided of granulomatous tissue and packed tightly with morselized cancellous femoral head allograft. All the revision acetabular components were implanted using cementless fixation. At a mean followup of 85 months (range, 60-118 months), the results of 103 of 108 hips (95%) were clinically and radiographically successful. The mean preoperative Harris hip score was 37 points, which improved to a mean of 91 points at the last followup. All cavitary defects had complete radiographic incorporation of the bone grafts except for two lesions. The results of this study indicate that morselized bone graft incorporates into cementless acetabular revisions. This treatment method provided a stable reconstruction in 98% of patients at a mean of 7 years followup. PMID- 15534550 TI - Retroacetabular osteolysis: when to operate? AB - Polyethylene liner exchange for retroacetabular osteolysis should be done before the shell becomes loose. The purpose of this study was to determine the radiographic quantity of osteolysis that will predict impending loosening of the cementless shell. Between 1992 and 2002, 46 cementless shells were revised at our institution for aseptic osteolysis. Radiographs and a computer-assisted technique were used to quantify osteolysis. Implant stability was confirmed intraoperatively. Of 26 stable and 20 loose shells, the average area of osteolysis on anteroposterior radiographs showed no significant difference, whereas lateral radiographs showed a difference. The percentage of shell circumference with associated osteolysis seen on anteroposterior and lateral radiographs showed a significant difference. Diagnostic criterion of 50% shell circumference associated with osteolysis on lateral films has a sensitivity of 0.84 and a specificity of 0.54, and on anteroposterior views, a sensitivity of 1.0 and a specificity 0.27 for predicting shell loosening. Percent of shell circumference with surrounding osteolysis seems to be more predictive of loosening than the area of osteolysis. When 50% of the shell circumference has osteolysis evident on anteroposterior or lateral radiographs, but preferably anteroposterior radiographs, liner exchange should be considered so that the exchange procedure is still possible, rather than allowing the osteolysis to increase and compromise shell fixation. PMID- 15534551 TI - Chemotherapy for bone sarcoma does not affect fertility rates or childbirth. AB - There have been unprecedented improvements in the survival of patients with bone sarcomas because of advances in chemotherapy during the past two decades. However, along with improved survival, there have been concerns of gonadal toxicity and fertility. These problems with chemotherapy are well documented for conditions like lymphomas and testicular cancers. There are few reports on fertility outcomes with chemotherapy for bone sarcomas. The purpose of this study was to review the rate of successful conceptions, pregnancy outcomes, incidence of birth defects, and fertility rates in young adults who had chemotherapy for high-grade bone sarcomas. A retrospective chart review of all eligible patients was done. No laboratory assessment of fertility was done. Fifteen of the 36 patients attempted conceptions, and all were successful [corrected] One miscarriage occurred and one medical termination of pregnancy was done because of a spinal metastasis. There were 13 successful full-term pregnancies with no birth defects. The conception rate was 1.6. Despite the common misconception of probable infertility, these patients can have high expectations to conceive, with uneventful childbirth and no birth defects in the newborns. Although counseling should include the possibility of infertility, patients also should be reminded of the high rate of success of having a normal conception and childbirth. PMID- 15534552 TI - 450 closed fractures of the distal third of the tibia treated with a functional brace. AB - Four hundred fifty closed fractures of the distal third of the tibial diaphysis, treated with a functional brace, are the subject of this study. Four (0.9%) of the fractures resulted in nonunion. The average healing time was 16.6 +/- 5.6 weeks, with a range from 10-40 weeks. The average final shortening was 5.1 +/- 4.8 mm with a range from 0-25 mm. Four hundred twenty four (94.2%) fractures healed with < 12 mm shortening. Initial shortening at the time of injury essentially was unchanged, from 4.4 +/- 4.5 mm to 4.4 +/- 3.9 mm final shortening. Axially unstable closed tibial fractures do not shorten beyond the initial shortening. Four-hundred five fractures (90.0%) healed with less than 8 degrees angular deformity in either the frontal or sagittal planes, and 302 (67.1%) healed with less than 5 degrees deformity in any plane. Overall, 391 fractures (87%) healed with shortening less than 12 mm and angulation in any plane less than 8 degrees . These degrees of angular deformity and shortening seem to compare favorably with those reported by other investigators using intramedullary nails. It seems that functional bracing is an effective method of treatment of a selected group of tibial fractures. PMID- 15534553 TI - Matrix metalloproteinases and their clinical applications in orthopaedics. AB - Imbalance in the expression of matrix metalloproteinases and their inhibitors contribute considerably to abnormal connective tissue degradation prevalent in various orthopaedic joint diseases such as rheumatoid arthritis and osteoarthritis. Matrix metalloproteinase expression has been detected in ligament, tendon, and cartilage tissues in the joint. They are known to contribute to the development, remodeling, and maintenance of healthy tissue through their ability to cleave a wide range of extracellular matrix substrates. Their role has been extended to cell growth, migration, differentiation, and apoptosis. In orthopaedics, their clinical applications constantly are being explored. The multiple steps in matrix metalloproteinase regulation offer potential targets for inhibition, useful in drug therapy. The correlation between matrix metalloproteinases and progression in joint erosion presents potential prognostic and diagnostic tools in rheumatoid arthritis. Matrix metalloproteinases also can be incorporated into scaffold design to control the degradation rate of engineered tissue constructs. This current review aims to summarize and emphasize the importance of matrix metalloproteinases and their natural inhibitors in the maturation of musculoskeletal tissue through matrix remodeling and, therefore, in the generation of a new clinical potential in orthopaedics. PMID- 15534554 TI - Wide contoured thigh cuffs and automated limb occlusion measurement allow lower tourniquet pressures. AB - We examined the amount of thigh tourniquet pressure that can be reduced from the typical 300 to 350 mm Hg by using a new automated plethysmographic limb occlusion pressure measurement technique. We also examined how much pressure could be reduced by using a wide contoured cuff compared with a standard cuff and if limb occlusion and systolic blood pressures were well correlated. Patients having surgery with a thigh tourniquet were randomized into two groups, one group having surgery with a standard cuff and the other with a wide cuff. Pressure was set at the automatically measured limb occlusion pressure plus a safety margin. Systolic blood pressure and quality of the bloodless field were recorded. The standard cuff maintained an acceptable bloodless field for 18 of 20 patients at an average pressure of 242 mm Hg, and the wide cuff was acceptable for 19 of 20 patients at an average of 202 mm Hg. One patient in each group had a poor bloodless surgical field at the initial pressure, and one patient in each group had a poor bloodless surgical field after a sharp rise in blood pressure during surgery. Systolic blood pressure was not correlated well enough to limb occlusion pressure to be used alone to set the optimum cuff pressure. The automated limb occlusion pressure technique and the wide contoured cuff reduced average pressure by 33-42% from typical pressures. PMID- 15534555 TI - The 2004 Marshall Urist award: delays until surgery after hip fracture increases mortality. AB - The objective of this study was to analyze whether a delay in time from admission until surgical treatment increased the mortality rate for patients with a closed hip fracture. We used the day of the week of admission as an instrumental variable to pseudorandomize patients. We analyzed 18,209 Medicare recipients who were 65 years of age or older and had surgical treatment for a closed hip fracture. Patients for whom the delay between admission and surgery was 2 days or more had a 17% higher chance of dying by Day 30. Using instrumental variables analysis, we found a similar 15% increased risk of mortality in patients with delays until surgery of 2 or more days. Based on these results, we found that a delay of 2 or more days significantly increased the mortality rate. This suggests that delay to surgery independently affects mortality, therefore additional study on the effect of smaller delays on outcome is needed. PMID- 15534556 TI - Forearm mass in an adolescent. PMID- 15534557 TI - A comparison of digital cameras: features essential for the orthopaedic surgeon. PMID- 15534559 TI - HIV clinical research in developing countries: challenges and priorities. PMID- 15534560 TI - Lessons for HIV from Tuskegee. AB - We are fortunate to live in an era in which research allows significant progress in the therapy of the major diseases that affect us. However, evaluating their efficacy and safety is expensive, laborious and involves the study of human subjects in an appropriate manner. The principles of such trials have a heterogeneous basis encompassing the principles of utilitarianism, distributive justice, the protection of individual rights and the investigators' commitment to patients. The Tuskegee study of untreated syphilis is the longest non-therapeutic experiment on human beings in medical history. It continues to cast its shadow on the relationship between black Americans and the broader biomedical community. Parallels and the relationship with the HIV pandemic are discussed. PMID- 15534561 TI - The European and developing countries clinical trails partnership. AB - The European and Developing Countries Clinical Trials Partnership (EDCTP) is a new venture between 14 states of the European Union (EU) and Norway to support the conduct of clinical trials of drugs and vaccines (and microbicides) against HIV/AIDS, malaria and tuberculosis in Africa, to develop the capacity to conduct such trials in African institutions, and to promote a more integrated approach to health research amongst European countries. It is funded for 5 years with a contribution of euros 200 million from the European Commission, matched by an equivalent sum that is spent directly by national research programmes of EU member states and Norway on activities that fall under the EDCTP remit. EDCTP seeks to be synergistic with other funding bodies supporting research on these diseases and will promote collaborative research, involving support from multiple funding agencies and harnessing and networking expertise across different African and European countries. PMID- 15534562 TI - Therapeutic vaccine trails in Thailand. AB - An effective therapeutic HIV vaccine will not only benefit the HIV-infected person but will also provide invaluable information on effective immune response in designing or selecting a preventive vaccine. A phase II double-blind controlled clinical trial using HIV-1 immunogen (Remune) was undertaken in Thailand in 1995 in conjunction with phase III approval in the USA. In most instances, immune response to the virus was induced. The results from this study led to the extension of further trials over the following 4 years. At present, the Thai Food and Drug Administration (FDA) is in the process of reviewing the files on all information to assess the effectiveness of this therapeutic vaccine as well as reviewing the proposal to conduct a phase III trial to further confirm previous efficacy results from the phase II trials. PMID- 15534563 TI - HIV and human leukocyte antigen association--the Indian scenario. AB - Genetic factors such as human leukocyte antigen (HLA) affect the infection and progression of HIV disease. The dependence of effective cytotoxic T lymphocyte (CTL) activity on HIV-specific T-helper responses has become increasingly evident and studies provide strong evidence that T-helper responses and the magnitude of CTL responses are closely linked. HLA haplotype A11-B35-Cw4 has been associated with rapid progression of AIDS. Our earlier studies had shown that HLA B*3520, B*1801 and Cw*1507 alleles were associated in western Indian HIV-1-infected AIDS patients. Further, it was observed that A*110101-B*3520-Cw*1507 haplotype was significantly increased among the AIDS patients. I compiled the frequency of these alleles and their molecular subtype distribution from 4333 individuals from India. The results showed that these high-risk alleles and their subtypes were significantly increased among selected castes or population groups (breeding isolates living in the plains) but lower in tribal groups (breeding isolates living in the hills), which revealed a high genetic diversity among Indians. My analysis shows that the genetic susceptibility and HLA allelic immune response among the HIV-infected AIDS individuals in India are different. PMID- 15534564 TI - Research as a path to wide-scale implementation of antiretroviral therapy in Africa. AB - Although some would deny the importance of research in resource-poor countries, the benefits of research to implementation of treatment for HIV infection are innumerable. These benefits include the development of infrastructure, training of staff, creation and validation of algorithms appropriate for the setting, and answering questions necessary for a safe and effective roll-out of therapy. This was true in the USA in 1986, 1 year after the antibody test for HIV was developed, and is true in Africa today. Shortly after the development of the HIV antibody test and before any antiretroviral therapy, few physicians or centres were willing to provide care for HIV patients and fewer had adequate facilities to do so. At that time it was not known how to make an adequate diagnosis of many of the opportunistic infections nor was there a clear idea of how to treat the patients. No-one knew either the best or most cost-effective method to prevent infections. Even as roll-out of therapy proceeded in early 1987 with the approval of zidovudine by the US Food and Drug Administration, physicians were clueless as to when to start treatment. With the addition of other medications in the armamentarium, clinicians began to make mistakes in their ignorance, adding on medications one at a time as they were approved, which led to accumulation of resistance mutations for a generation of patients. These mutations were transmitted to partners and children. What single-handedly helped advance treatment in the USA and Europe in the 1980s was the willingness of respective governing authorities to create clinical research groups not only to develop new drugs but to help create cost-effective ways to use them. All the current treatment guidelines were developed from that research. Over the years these research groups provided care, including medications, laboratory tests and physician and nurse time, for thousands of patients. Medical centres, where these indigent patients were receiving their care, were encouraged to open their doors, creating state of the art clinics and inpatient wards. A generation of clinicians was trained at these research centres where the bulk of US HIV patients were treated. They provided care as they were conducting research. The ability of resource-poor countries to deliver large-scale roll-out plans is dependent on the development of leadership and skills to implement the programmes. South Africa, despite a delay in initiating a national treatment programme, is an example of a country where the research conducted in the period 1996 to 2004 has enabled a skilled set of clinicians, pharmacists and paramedical staff to provide leadership in the scale up of antiretroviral therapy programmes. Guideline development, training and implementation have been led by treatment experts who learned their skills in the research arena. PMID- 15534565 TI - XV International AIDS Conference, Bangkok, Thailand, 11-16 July 2004: expanding research in developing countries. PMID- 15534566 TI - Tablet splitting. PMID- 15534567 TI - COX2-alternatives and GI protection. PMID- 15534568 TI - Systemic reactions to imiquimod (Aldara). PMID- 15534569 TI - The primary/secondary care interface in gastroenterology. PMID- 15534570 TI - Top down or bottom up?-How should GPs cultivate their higher professional education needs in mental healthcare? PMID- 15534571 TI - Complications of the intrauterine device in nulliparous and parous women. AB - OBJECTIVES: The intrauterine device (IUD) is still related to pelvic inflammatory disease (PID), pregnancy, expulsion, perforation and menstrual problems, particularly in nulliparous women. We aimed to study the complications and symptoms of the intrauterine device in general practice, particularly in nulliparous women. METHODS: We used a retrospective cohort study in four general practices participating in the Nijmegen Continuous Morbidity Registration. Selected women had one or more IUDs inserted between 1981 and 2000. Data on complications, symptoms and removal of the IUD were obtained from the medical records. RESULTS: 461 women were included, 129 nulliparous and 332 parous women. Users of copper IUDs had a rate of PID of 3.5 per 1000 women-years, rates of (ectopic) pregnancy of 0.6 to 1.1% per year and rates of expulsion of 0 to 1.2% per year. Rates of expulsion for the levonorgestrel-releasing IUDs were 0 to 0.2% per year. Nulliparous women did not show more complications than parous women. Menstrual problems were frequent among users of copper and levonorgestrel releasing IUDs. One third of the IUDs were removed within the first year after insertion. There was no significant excess of IUD removal among nulliparous women compared with parous women. Main reasons for removal were 'menstrual problems' and 'contraception no longer necessary'. CONCLUSION: Both copper and levonorgestrel-releasing IUDs are safe and highly effective contraceptives, which can adequately be inserted and monitored by general practitioners in nulliparous and parous women. PMID- 15534572 TI - Spanish primary healthcare physicians' knowledge of resistance of respiratory germs and antimicrobial treatment in acute bronchitis. AB - OBJECTIVES: To determine what Spanish primary healthcare physicians consider the rates of pneumococcal resistance to be, the type of antimicrobial treatment they would prescribe on suspicion of high resistance and ascertain in which cases of acute bronchitis these physicians would initiate antibiotic treatment. METHODS: A cross-sectional, observational study was performed with a questionnaire distributed to 1368 physicians on their opinion on the degree of resistance of pneumococci to penicillins and macrolides in Spain. They were also asked which treatment they would recommend in penicillin-resistant pneumococcal infection and to indicate their degree of agreement regarding the use of antibiotics in different clinical situations of acute bronchitis. RESULTS: 843 physicians completed and returned the questionnaire, a response rate of 59.1%. Of the 750 physicians who answered the question concerning pneumococcal resistance, 312 stated resistance to be >30% (41.6%) while only 90 felt it to be <15% (12%). 153 physicians reported resistance to macrolides to be >30% (23%). In cases with suspicion of penicillin-resistant pneumococci, the clinicians stated they would prescribe quinolones (321, 40.3%) and macrolides (251, 31.5%). Of the 784 physicians who answered the questions on bronchitis, 662 preferred antibiotic therapy in acute bronchitis with purulent sputum (84.6%), 610 on uncertain diagnosis (77.8%), 569 in patients with fever over 38 degrees C (72.6%) and 210 on patient request (26.8%). CONCLUSION: Primary healthcare physicians in Spain are more concerned about pneumococcal resistance to penicillins than to macrolides although 35% of the strains isolated are resistant to the latter while only 10% are resistant to aminopenicillins. Furthermore, many physicians believe macrolides to be effective against pneumococcal infections. Despite their lack of efficacy in the treatment of acute bronchitis, many physicians prescribe antibiotics, mainly in cases with purulent expectoration, thus justifying the high consumption of antimicrobial agents in this disease. PMID- 15534573 TI - Arriving at the postmodern medical consultation. AB - The analysis of the medical consultation is characterised by mainly prescriptive attempts to recommend 'best practice'. As the role of the individual in society has gained prominence, the power relationships in medical practice have had to change to reflect the increasing recognition of autonomy and self-determination. Medical discourse is at a junction, having to relinquish authoritarianism and grapple with the concept of sharing information and decisions in an area where complex and uncertain data exist, albeit often without full disclosure. The writing of Foucault and Lyotard and the concept of postmodernism fit well with the idea that the consultation between doctors and patients is increasingly becoming a contested space that occupies multiple voices, such as that of the media, the pharmaceutical industry, government-led guidelines and that of the profession. Creating the circumstances and the means for creating an effective dialogue in the postmodern consultation is the prime task for physicians. PMID- 15534574 TI - The long march: the development of academic general practice in the UK and Ireland. AB - Academic general practice and primary care is thriving in both the UK and Ireland, as judged by the content and vibrancy of the main annual scientific meetings of the discipline in the two countries. Problems and challenges abound, but the long-term trend is one of steady growth, with gradual increases in capacity and activity. Contributions to undergraduate medical education and to multidisciplinary professional development, and achievements in the UK University Research Assessment Exercise (RAE) show what the discipline is capable of. A major challenge remains the creation and funding of appropriate academic career structures in academic general practice and primary care, removing the current financial disincentives, uncertainties and problems in acquiring both clinical and academic experience. With increased numbers of clinical and non-clinical academics working in general practice and primary care, the discipline is set to play an important role in health service development in both the UK and Ireland. PMID- 15534575 TI - Panorama of diagnoses in the primary healthcare setting in Cyprus-data from a pilot study. PMID- 15534576 TI - Introducing general practice in urban Greece: focus on morbidity profile. PMID- 15534577 TI - Peripheral arterial disease: the gap between common practice and the Dutch College of General Practitioners guideline. PMID- 15534578 TI - Underuse of aspirin therapy among patients with type 2 diabetes mellitus attending primary care in Greece. PMID- 15534580 TI - Ever increasing need for family physicians! PMID- 15534581 TI - Great but unrealistic expectations. PMID- 15534579 TI - Control of vascular and renal risk factors in diabetic patients-results of the Barbanza Diabetes Educational Programme. PMID- 15534583 TI - Tissue specific expression of alternative splice forms of human cyclic nucleotide gated channel subunit CNGA3. AB - PURPOSE: The cyclic nucleotide gated cation channel subunit, CNGA3, (also known as the cone alpha CNG subunit), plays a vital role in signal transduction of human cone cells. Owing to its well established role in vision, the human CNGA3 isoform studied thus far was cloned from retinal tissue. However, non-human homologs of CNGA3 have been cloned from a variety of other tissues including kidney, heart, pineal gland, adrenal gland and testes. Among these, alternative splice forms of CNGA3 have been identified. The objective of this study was to explore alternative splice forms of human CNGA3 and determine their distribution among various human tissues. METHODS: RT-PCR was used to amplify full length open reading frames of CNGA3 from human testes RNA and to detect and distinguish among splice forms in 23 tissues. DNA sequencing was used to characterize full length splice forms and to confirm the identity of RT-PCR products from a number of tissues. RESULTS: Two new full length alternatively spliced forms of hCNGA3 (referred to as Variant 2 and 3) were isolated. These splice variants are different from the cone hCNGA3 (Variant 1) in that they lack exon 5. In addition, they differ from each other in that Variant 3 contains an extra exon that originates from the intron preceding exon 4. We demonstrate that CNGA3 transcripts are detectable in all 23 human tissues examined. In all tissues, except retina, Variant 2 specific PCR products had the brightest band intensity. In retina, the band from Variant 1 (containing exon 5) was most intense. In fact, among all tissues examined, Variant 1 can only be detected strongly in the retina. CONCLUSIONS: The extensive distribution of CNGA3 and the tissue specific expression of alternative splice forms indicate widespread and diverse roles for CNGA3. The unique expression of Variant 1 in the retina implies a significance to the amino acids encoded by exon 5 that may be necessary for the function of CNGA3 in human cone cells. PMID- 15534584 TI - The damaging effect of UV-C irradiation on lens alpha-crystallin. AB - PURPOSE: To evaluate the effect of UV-C irradiation on the structural properties of alpha-crystallin and its chaperone activity. METHODS: alpha- and betaL crystallins were isolated from bovine lenses using gel chromatography. The purified alpha-crystallin was subjected to UV-C irradiation (254 nm; 1, 2, 5, 10, 20, 50 J/cm2). We measured the tryptophan fluorescence, circular dichroism (CD) spectroscopy in the far UV, and the chaperone activity of both irradiated and non irradiated alpha-crystallin. RESULTS: The tryptophan fluorescence of alpha crystallin decreased, whereas the N-formylkynurenine fluorescence increased markedly with increasing doses of UV-C irradiation. Both the oxidation of Met1 and the racemization of Asp151 of alphaA-crystallin increased at a dose of 1-2 J/cm2 and then gradually decreased. The CD spectrum showed that the secondary structure of alpha-crystallin altered with increasing radiation dose, and almost all of the beta-sheet structure was lost at doses above 50 J/cm2. The chaperone activity of alpha-crystallin irradiated with doses under 5 J/cm2 remained intact. However, it was reduced to only 40% after irradiation at 10 J/cm2. CONCLUSIONS: Our study suggests that photo-oxidation of tryptophan residues in alpha crystallin may be one of the events that affects the three-dimensional packing array and chaperone activity of this lens protein. PMID- 15534585 TI - Genetic correlations among texture characteristics in the human iris. AB - PURPOSE: To estimate the magnitude of genetic correlations among five general textural characteristics of the human iris. METHODS: Color photographs of iris were available from 100 monozygotic and 99 dizygotic twin pairs. Comparative scales were constructed based on ratings of the subjects' left iris. To explore the genetic and environmental covariation among frequency of Fuchs' crypts, frequency of pigment dots, iris color, the extension, and distinction of Wolfflin nodules, and contraction furrows, a structural equation model with Cholesky decomposition was applied to variance-covariance matrices for monozygotic (MZ) and dizygotic (DZ) pairs. RESULTS: Significant genetic correlations fell between 0.22 and 0.44 and accounted almost entirely for the phenotypic correlations among the iris characteristics. No evidence for individual specific environmental effects in common to the characteristics was found. CONCLUSIONS: The modest genetic correlations indicate that there is little overlap in the genetic influence for these characteristics. Candidate genes with embryological and histological expression patterns in the eye could potentially influence the iris characteristics' variability. PMID- 15534587 TI - Primary tumours of the optic nerve and its sheath. AB - PURPOSE: To describe the clinical, neuroimaging, and pathologic features of primary tumours of the optic nerve and its sheath. METHODS: Review of published cases and personal series. RESULTS: The most common primary tumour of the optic nerve is the benign glioma. This low-grade astrocytoma usually can be followed without intervention. Progression of visual symptoms and signs may necessitate either surgery to remove the tumour or radiation therapy. The most common tumour of the optic nerve sheath is the meningioma. The optimum treatment for this lesion is stereotactic or three-dimensional conformal fractionated radiation therapy, which generally results in stabilization or improvement in vision. A variety of other primary tumours may mimic, in both manifestations and imaging appearance, the more common glioma or meningioma. In such cases, the correct diagnosis may not be made until a biopsy is performed or the nerve is removed. CONCLUSION: Primary tumours of the optic nerve and its sheath are not uncommon. Diagnosis can often but not always be made by the results of a complete examination combined with imaging studies, particularly CT scanning and MR imaging. Management depends on the presumed or histologically verified nature of the tumour. PMID- 15534588 TI - Congenital disorders of the optic nerve: excavations and hypoplasia. AB - The principal congenital abnormalities of the optic disc that can significantly impair visual function are excavation of the optic disc and optic nerve hypoplasia. The excavated optic disc abnormalities comprise optic disc coloboma, morning glory syndrome, and peripapillary staphyloma. Optic nerve hypoplasia manifests as a small optic nerve, which may or may not be accompanied by a peripapillary ring (the double ring sign). In addition, the optic disc cupping, which occurs as a sequel to some cases of periventricular leucomalacia, can arguably be classified as a type of optic nerve hypoplasia. All of these conditions can be unilateral or bilateral and can impair visual function mildly or severely. It is essential that children with poor vision due to any of these conditions are managed by treating refractive errors, giving occlusion therapy in selected cases, and optimising the conditions at home and at school in an attempt to ensure that impaired vision does not impede development or education. PMID- 15534589 TI - Gene therapy for optic nerve disease. AB - PURPOSE: There has been recent interest in the potential use of gene therapy techniques to treat ocular disease. In this article, we consider the optic nerve diseases that are potentially most amenable to gene therapy. METHODS: We discuss the recent success of gene transfer experiments in animal models of glaucoma, optic neuritis, Leber's hereditary optic neuropathy (LHON), and optic nerve transection, and we assess the possibility of using similar techniques to treat human disease in the future. RESULTS: We have achieved highly efficient transfection of retinal ganglion cells in a rat model of glaucoma following a single intravitreal injection of adeno-associated virus (AAV). In our model, we have found that AAV-mediated gene therapy with brain-derived neurotrophic factor has a significant neuroprotective effect compared to saline or control virus injections. Guy and co-workers have successfully used AAV-mediated gene therapy to replace the defective mitochondrial enzyme subunit in cells derived from human patients with LHON. Gene therapy techniques have also shown promise in animal models of optic neuritis and optic nerve trauma. CONCLUSIONS: Human diseases with single-gene defects such as LHON may soon be treated successfully by gene therapy, assuming that vectors continue to improve and are well tolerated in the human eye. Other optic nerve diseases such as glaucoma that do not have a single gene defect may also benefit from gene therapy to enhance RGC survival. In all cases, the risks of treatment will need to be balanced against the potential benefits. PMID- 15534590 TI - Intraocular surgery for optic nerve disorders. AB - This article discusses two current indications for intraocular surgery for optic nerve conditions. The first topic is macular detachment in association with congenital optic disc pit, and includes a review of the theories of pathogenesis and management options including our own experience of treatment with vitrectomy and gas tamponade. In addition, the histopathological findings of a new case of optic disc pit with serous macular detachment are presented, adding to the rare examples previously reported. The second subject is a radial optic neurotomy, a recently advocated treatment for central retinal vein occlusion, which, despite its controversial nature, is gaining in popularity. PMID- 15534591 TI - Animal models of optic nerve diseases. PMID- 15534592 TI - Optic nerve and neuroprotection strategies. AB - BACKGROUND: Experimental studies have yielded a wealth of information related to the mechanism of ganglion cell death following injury either to the myelinated ganglion cell axon or to the ganglion cell body. However, no suitable animal models exist where injury can be directed to the optic nerve head region, particularly the unmyelinated ganglion cell axons. The process of relating the data from the various animal models to many different types of optic neuropathies in man must, therefore, be cautious. RESULTS: Extensive studies on the isolated optic nerve have yielded valuable information on the way white matter is affected by ischaemia and how certain types of compounds can attenuate the process. Moreover, there are now persuasive data on how ganglion cell survival is affected when the ocular blood flow is reduced in various animal models. As a consequence, the molecular mechanisms involved in ganglion cell death are fairly well understood and various pharmacological agents have been shown to blunt the process when delivered before or shortly after the insult. CONCLUSIONS: A battery of agents now exist that can blunt animal ganglion cell death irrespective of whether the insult was to the ganglion cell body or the myelinated axon. Whether this information can be applied for use in patients remains a matter of debate, and major obstacles need to be overcome before the laboratory studies may be applied clinically. These include the delivery of the pharmacological agents to the site of ganglion cell injury and side effects to the patients. Moreover, it is necessary to establish whether effective neuroprotection is only possible when the drug is administered at a defined time after injury to the ganglion cells. This information is essential in order to pursue the idea that a neuroprotective strategy can be applied to a disease like glaucoma, where ganglion cell death appears to occur at different times during the lifetime of the patient. PMID- 15534593 TI - Stem cells to replace the optic nerve. AB - Methods that exist now and that might be developed are suggested to replace retinal ganglion cells and their axons in the optic nerve, ultimately to re establish functional vision in eyes blind from glaucoma. PMID- 15534594 TI - Circulation and axonal transport in the optic nerve. AB - Retinal ganglion cells are the output cells of the retina whose axons are under considerable metabolic stress in both health and disease states. They are highly polarised to ensure that mitochondria and enzymes involved in the generation of ATP are strategically concentrated to meet the local energy demands of the cell. In passing from the eye to the brain, axons are protected and supported by glial tissues and the blood supply of the optic nerve head is regulated to maintain the supply of oxygen and nutrients to the axons. In spite of this, the optic nerve head remains the point at which retinal ganglion cell axons are most vulnerable to the effects of increased intraocular pressure or ischaemia. Considerable work has been undertaken in this area to advance our understanding on the pathophysiology of axon damage and to develop new strategies for the prevention of retinal ganglion cell death. PMID- 15534595 TI - A history of the optic nerve and its diseases. AB - We will trace the history of ideas about optic nerve anatomy and function in the Western world from the ancient Greeks to the early 20th century and show how these influenced causal theories of optic nerve diseases. Greek and Roman humoral physiology needed a hollow optic nerve, the obstruction of which prevented the flow of visual spirit to and from the brain and resulted in blindness. Medieval physicians understood that the presence of a fixed dilated pupil indicated optic nerve obstruction, preventing the passage of visual spirit, and that cataract surgery in such cases would not restore sight. During the Renaissance, the organ of vision was transferred from the lens to the optic nerve, which was generally believed to be on the axis of the eye. The acuity of central vision (at the optic disc) was explained by the concentration of visual spirit where the optic nerve met the retina. The growth of anatomy and influence of mechanical philosophy from the 17th century led to visual spirit being replaced with the concept of nerve force, which later became associated with electricity travelling along nerve fibres. This coincided with discourse about the nature of the nervous system and a shift in orientation from understanding illness holistically in terms of an individual's humoral imbalance to the concept of organ-based diseases. Both the microscope and the ophthalmoscope allowed visualisation of the optic nerve, but problems of interpretation persisted until conceptual transformations in medical science were made. PMID- 15534596 TI - Functions of optic nerve glia: axoglial signalling in physiology and pathology. AB - An early concept of glial function envisaged them as passive and unexcitable structural elements, much like the connective tissues of organs in the periphery. It is now known that glia have a widespread range of physiological roles and react to all forms of pathological insult. This paper reviews the major functions of oligodendrocytes and astrocytes, the main types of glia in the optic nerve, and examines novel NG2-glia, otherwise known as oligodendrocyte progenitor cells (OPCs). The major function of oligodendrocytes is to produce the myelin sheaths that insulate CNS axons, but they also have important roles in the establishment of nodes of Ranvier, the sites of action potential propagation, and axonal integrity. Astrocytes have multiple physiological and pathological functions, including potassium homeostasis and metabolism, and reactive astrogliosis in response to CNS insults. The bulk of NG2-glia are postmitotic complex cells, distinct from OPCs, and respond to any insult to the CNS by a rapid and stereotypic injury response. This may be their primary unction, but NG2-glia, or a subpopulation of NG2-expressing adult OPCs, also provide remyelinating oligodendrocytes following demyelination. Oligodendrocytes, astrocytes, and NG2 glia all contact axons at nodes of Ranvier and respond to glutamate, ATP, and potassium released during axonal electrical activity. Glutamate and ATP evoke calcium signalling in optic nerve glia and have dual roles in physiology and pathology, coupling glial functions to axonal activity during normal activity, but enhanced activation induces an injury response, as seen following injury, demyelination, and ischaemia. PMID- 15534597 TI - Traumatic optic neuropathy. PMID- 15534598 TI - Molecular genetic basis of primary inherited optic neuropathies. AB - AIM: To review the molecular genetic basis of primary inherited optic neuropathies. METHODS: Medline and Embase search. RESULTS: Inherited optic neuropathies are a genetically diverse group of disorders that present with reduced visual acuity and the clinical appearance of optic atrophy. The inherited optic neuropathies may be sporadic or familial, in which case the mode of inheritance may be Mendelian (autosomal dominant, autosomal recessive, X-linked recessive) or non-Mendelian (mitochondrial). Two genes for dominantly inherited optic atrophy have been mapped (OPA1 and OPA4), of which the gene has been identified in one (OPA1). A gene for recessive optic atrophy (OPA3) has also been identified. X-linked optic atrophy (OPA2) has been mapped but to date no gene has been identified. Mutations in mitochondrial DNA have been identified in Leber's hereditary optic neuropathy. CONCLUSIONS: Mutations in genes from both the nuclear and mitochondrial genomes appear to be responsible. Mitochondrial dysfunction, in the broadest sense, is emerging as central to the pathogenesis of this group of conditions. PMID- 15534599 TI - Electrophysiological assessment of optic nerve disease. AB - The electrophysiological findings in optic nerve and primary ganglion cell dysfunction are reviewed. The value of the pattern reversal visual-evoked potential (VEP) in the diagnosis of optic nerve disease, and the pattern appearance VEP in the demonstration of the intracranial misrouting associated with albinism, are discussed. The pattern electroretinogram (PERG) is used in the direct assessment of ganglion cell function. The use of PERG or multifocal electroretinography (mfERG), to enable the distinction between VEP delay due to optic nerve disease and that due to macular dysfunction, is described. PMID- 15534600 TI - Hereditary optic neuropathies. AB - AIMS: To provide a clinical update on the hereditary optic neuropathies. METHODS: Review of the literature. RESULTS: The hereditary optic neuropathies comprise a group of disorders in which the cause of optic nerve dysfunction appears to be hereditable, based on familial expression or genetic analysis. In some hereditary optic neuropathies, optic nerve dysfunction is typically the only manifestation of the disease. In others, various neurologic and systemic abnormalities are regularly observed. CONCLUSION: The most common hereditary optic neuropathies are autosomal dominant optic atrophy (Kjer's disease) and maternally inherited Leber's hereditary optic neuropathy. We review the clinical phenotypes of these and other inherited disorders with optic nerve involvement. PMID- 15534601 TI - Clinical assessment of optic nerve disorders. AB - Optic nerve disorders range from the manifestations of life-threatening intracranial or systemic disease, to minor congenital anomalies. Careful clinical assessment, which relies upon a thorough evaluation of symptoms as well as signs, is essential for effective and timely investigation and treatment. Examination methods and pitfalls to be avoided are discussed. PMID- 15534602 TI - Optic nerve disorders: role of canal and nerve sheath decompression surgery. AB - Optic nerve sheath decompression (ONSD) maintains a role in the management of visual loss complicating papilloedema in selected patients primarily with idiopathic intracranial hypertension. The evidence base for this intervention is reviewed and audit data on visual outcomes for patients with acute, chronic, and atrophic forms of papilloedema are contrasted. Optic canal decompression has a role in the management of compressive optic neuropathies complicating mass lesions arising from paranasal sinuses and intracranially and can be achieved by transethmoidal, transbasal, and open craniotomy routes. The evidence base supporting this intervention in traumatic optic neuropathy and in primary bone disease causing canal stenosis (in particular fibrous dysplasia) is reviewed where the indications are more controversial. PMID- 15534603 TI - Pupil assessment in optic nerve disorders. AB - BACKGROUND: The normal pupillary constriction to light is an involuntary reflex that can be easily elicited and observed without specialized equipment or discomfort to the patient. Attenuation of this reflex in optic nerve disorders was first described 120 years ago. Since then, pupil examination has become a routine part of the assessment of optic nerve disease. CLINICAL TECHNIQUES: The original cover/uncover test compares pupillomotor drive in the two eyes, but requires two working pupils and is relatively insensitive. The swinging flashlight test is now the standard clinical tool to detect pupillomotor asymmetry. It requires only one working pupil, is easily quantified, and has high sensitivity in experienced hands, but interpretation of the results needs care. Measurement of the pupil cycle time is the only clinical test that does not rely on comparison with the fellow eye, but it can only be measured in mild to moderate optic nerve dysfunction, is more time consuming, and less sensitive. LABORATORY TECHNIQUES: Infrared video pupillography allows recordings to be made of the pupil responses to full-field or perimetric light stimulation under tightly controlled conditions with a high degree of accuracy. Frustratingly, there is a wide range in reflex gain in normal subjects limiting its usefulness unless comparison is made with the fellow eye or stimulation of unaffected adjacent areas of the visual field. CORRELATION WITH OTHER TESTS: In general, pupillomotor deficit shows good correlation with visual field deficit. However, some diseases of the optic nerve are associated with relative sparing either of pupil function or visual function implying that pupil tests and psychophysical tests may assess function in different subpopulations of optic nerve fibres. Less is known of the relationship between pupil measurements and electrodiagnostic tests. USES IN CLINICAL PRACTICE: Pupil assessment is invaluable when distinguishing functional from organic visual loss. Its usefulness in distinguishing between different causes of optic neuropathy and as a prognostic sign is gradually emerging. PMID- 15534604 TI - Mechanisms of axon-glial injury of the optic nerve. AB - The central concept underlying ideas on the pathogenesis of multiple sclerosis is that inflammatory events cause acute injury of axons and myclin. The phases of symptom onset, recovery, persistence, and progression in multiple sclerosis can be summarized as functional impairment with intact structure due to direct effects of inflammatory mediators; demyelination and axonal injury with recovery through plasticity and remyelination; and chronic axonal loss due to failure of enduring remyelination from loss of trophic support for axons normally provided by cells of the oligodendrocyte lineage. Cell death may occur in response to a state of injury from which protection would be anticipated under more favourable neurobiological conditions. Conversely, optimal growth factor environment may save cells from otherwise lethal events occurring at the cell membrane. Hence, in the context of brain inflammation, there is an inseparable interplay between immunological and neurobiological contributions to tissue injury. PMID- 15534605 TI - Posterior ischaemic optic neuropathy: clinical features, pathogenesis, and management. AB - PURPOSE: To investigate and present a comprehensive account of the clinical features, pathogenesis, and management of posterior ischaemic optic neuropathy (PION). METHODS: This retrospective study is based on 53 consecutive eyes of 42 patients with PION seen in my clinic since 1973, who fulfilled the inclusion criteria. They were systematically evaluated, treated, and followed by me. All patients had initially detailed ophthalmic evaluation of the anterior and posterior segments, including visual field with Goldmann perimeter and fluorescein fundus angiography. All patients aged 50 years and older were also investigated for giant cell arteritis (GCA). Every attempt was made to rule out other causes of visual loss. Follow-up evaluation was similar to the initial evaluation except angiography. Aetiologically, PION can be divided into three types: arteritic due to GCA, nonarteritic not due to GCA, and surgical following a surgical procedure. Steroid therapy was given to only those nonarteritic PION patients who opted to try that, but was given to all arteritic PION patients. RESULTS: PION was nonarteritic in 28 patients (35 eyes), arteritic in 12 (14 eyes), and surgical in three (four eyes). Visual acuity varied between 20/20 and no light perception--it was count fingers or less in 19 of 35 eyes with nonarteritic PION, four of 14 in arteritic, and all four with surgical PION. The most common visual field defect was central visual loss, alone or in combination with other types of visual field defects. Initially, optic disc and fundus showed no abnormality but the disc usually developed pallor in about 6-8 weeks. Aggressive treatment with high-dose systemic steroid during the very early stages of nonarteritic PION produced significant improvement of visual acuity as well as visual fields, but not so in arteritic or surgical PION. However, some spontaneous visual improvement also occurred in some untreated nonarteritic PION cases. CONCLUSIONS: PION is a distinct clinical entity but should be diagnosed only after exclusion of all other causes of visual loss. In all patients older than 50, GCA must be ruled out. There is usually marked visual loss, with central field defect being the most common. The study suggests that high-dose steroid therapy in nonarteritic PION, soon after the onset of visual loss, resulted in significant visual improvement compared to the untreated cases, but not in arteritic and surgical PION. PMID- 15534606 TI - Image analysis of optic nerve disease. AB - Existing methodologies for imaging the optic nerve head surface topography and measuring the retinal nerve fibre layer thickness include confocal scanning laser ophthalmoscopy (Heidelberg retinal tomograph), optical coherence tomography, and scanning laser polarimetry. For cross-sectional screening of patient populations, all three approaches have achieved sensitivities and specificities within the 60 80th percentile in various studies, with occasional specificities greater than 90% in select populations. Nevertheless, these methods are not likely to provide useful assistance for the experienced examiner at their present level of performance. For longitudinal change detection in individual patients, strategies for clinically specific change detection have been rigorously evaluated for confocal scanning laser tomography only. While these initial studies are encouraging, applying these algorithms in larger numbers of patients is now necessary. Future directions for these technologies are likely to include ultra high resolution optical coherence tomography, the use of neural network/machine learning classifiers to improve clinical decision-making, and the ability to evaluate the susceptibility of individual optic nerve heads to potential damage from a given level of intraocular pressure or systemic blood pressure. PMID- 15534607 TI - Long-term population-based risks of second malignant neoplasms after childhood cancer in Britain. AB - In a population-based, retrospective cohort study of 16 541 3-year survivors of childhood cancer treated in Britain up to the end of 1987, 278 second malignant neoplasms (SMNs) were identified against 39.4 expected giving a standardised incidence ratio (SIR) of 6.2. The overall cumulative risk of an SMN by 25 years from 3-year survival from childhood cancer was 4.2%. Analysis of the cohort of nonretinoblastoma childhood cancers combined revealed a significant decline in SIR of SMN with increasing duration of follow-up. There was a greater risk of developing a SMN, particularly secondary acute myeloid leukaemia, in those diagnosed with childhood cancer from 1980 onwards. However, on multivariate modeling, this was not an independent risk factor. There was significant heterogeneity (P<0.001) in SIR of SMN across different treatment groups, the greatest risk observed in the group exposed to both radiotherapy and chemotherapy. The risks of SMN observed were comparable with those in other population-based studies. While the decline in SIR with duration of follow-up and the small excess numbers of cancers observed over later decades after diagnosis are reassuring, the high excess risk, particularly of leukaemia, associated with recent more intense therapy is of concern. PMID- 15534608 TI - Psychological costs of inadequate cervical smear test results. AB - The purpose of the study is to investigate, for the first time, the psychological impact of an inadequate smear test result. A prospective questionnaire design was used, and the setting was a single English cervical screening laboratory. Two groups of women receiving either a normal test result (n=226) or either a first or nonconsecutive inadequate smear test result (n=180) participated. The main outcome measures included State anxiety (STAI) and concern about test result, assessed within 4 weeks of receipt of results, and attendance for a repeat cervical smear within 3 months. Compared to women with normal test results, women with inadequate smear test results had higher state anxiety (P=0.025), were more concerned about their results (P<0.001), perceived themselves to be at higher risk of cervical cancer (P=0.016), and felt less satisfied with the information they had received about their test results (P<0.001). The only predictor of attendance for a repeat smear test following an inadequate smear test result was state anxiety (P=0.011): nonattenders had higher levels of state anxiety in response to their initial test results. In conclusion, in this first study to assess the psychological impact of receiving an inadequate smear test result, we have shown that it raises state anxiety and concern to levels similar to those found in women with abnormal smear test results. Of particular concern is that anxious women are less likely to attend for a repeat smear test within the recommended time frame. Given the millions of women each year receiving this test result, research is now needed to ascertain how the anxiety associated with this result can be avoided. PMID- 15534609 TI - Epigenetic inactivation of SLIT3 and SLIT1 genes in human cancers. AB - In Drosophila, the Slit gene product, a secreted glycoprotein, acts as a midline repellent to guide axonal development during embryogenesis. Three human Slit gene orthologues have been characterised and recently we reported frequent promoter region hypermethylation and transcriptional silencing of SLIT2 in lung, breast, colorectal and glioma cell lines and primary tumours. Furthermore, re-expression of SLIT2 inhibited the growth of cancer cell lines so that SLIT2 appears to function as a novel tumour suppressor gene (TSG). We analysed the expression of SLIT3 (5q35-34) and SLIT1 (1q23.3-q24) genes in 20 normal human tissues. Similar to SLIT2 expression profile, SLIT3 is expressed strongly in many tissues, while SLIT1 expression is neuronal specific. We analysed the 5' CpG island of SLIT3 and SLIT1 genes in tumour cell lines and primary tumours for hypermethylation. SLIT3 was found to be methylated in 12 out of 29 (41%) of breast, one out of 15 (6.7%) lung, two out of six (33%) colorectal and in two out of (29%) glioma tumour cell lines. In tumour cell lines, silenced SLIT3 associated with hypermethylation and was re-expressed after treatment with 5-aza-2'-deoxycytidine. In primary tumours, SLIT3 was methylated in 16% of primary breast tumours, 35% of gliomas and 38% of colorectal tumours. Direct sequencing of bisulphite-modified DNA from methylated tumour cell lines and primary tumours demonstrated that majority of the CpG sites analysed were heavily methylated. Thus, both SLIT2 and SLIT3 are frequently methylated in gliomas and colorectal cancers, but the frequency of SLIT3 methylation in lung and breast cancer is significantly less than that for SLIT2. We also demonstrated SLIT1 promoter region hypermethylation in glioma tumour lines (five out of six; 83%), the methylation frequency in glioma tumours was much lower (two out of 20; 10%). Hence, evidence is accumulating for the involvement of members of the guidance cues molecules and their receptors in tumour development. PMID- 15534610 TI - Function and quality-of-life of survivors of pelvic and lower extremity osteosarcoma and Ewing's sarcoma: the Childhood Cancer Survivor Study. AB - Limb-sparing surgeries have been performed more frequently than amputation based on the belief that limb-sparing surgeries provide improved function and quality of-life (QOL). However, this has not been extensively studied in the paediatric population, which has unique characteristics that have implications for function and QOL. Using the Childhood Cancer Survivor Study, 528 adult long-term survivors of pediatric lower extremity bone tumours, diagnosed between 1970 and 1986, were contacted and completed questionnaries assessing function and QOL. Survivors were an average of 21 years from diagnosis with an average age of 35 years. Overall they reported excellent function and QOL. Compared to those who had a limb sparing procedure, amputees were not more likely to have lower function and QOL scores and self-perception of disability included general health status, lower educational attainment, older age and female gender. Findings from this study suggest that, over time, amputees do as well as those who underwent limb-sparing surgeries between 1970 and 1986. However, female gender, lower educational attainment and older current age appear to influence function, QOL and disability. PMID- 15534611 TI - Inherent growth advantage of (pre)malignant hepatocytes associated with nuclear translocation of pro-transforming growth factor alpha. AB - The pro-peptide of transforming growth factor alpha (proTGFalpha) was recently found in hepatocyte nuclei preparing for DNA replication, which suggests a role of nuclear proTGFalpha for mitogenic signalling. This study investigates whether the nuclear occurrence of the pro-peptide is involved in the altered growth regulation of (pre)malignant hepatocytes. In human hepatocarcinogenesis, the incidence of proTGFalpha-positive and replicating nuclei gradually increased from normal liver, to dysplastic nodules, to hepatocellular carcinoma. ProTGFalpha positive nuclei almost always were in DNA synthesis. Also, in rat hepatocarcinogenesis, proTGFalpha-positive nuclei occurred in (pre)malignant hepatocytes at significantly higher incidences than in unaltered hepatocytes. For functional studies unaltered (GSTp(-)) and premalignant (GSTp(+)) rat hepatocytes were isolated by collagenase perfusion and cultivated. Again, DNA synthesis occurred almost exclusively in proTGFalpha-positive nuclei. GSTp(+) hepatocytes showed an approximately 3-fold higher frequency of proTGFalpha-positive nuclei and DNA replication than GSTp(-) cells. Treatment of cultures with the mitogen cyproterone acetate (CPA) elevated the incidence of proTGFalpha-positive nuclei and DNA synthesis in parallel. Conversely, transforming growth factor beta1 (TGFbeta1) lowered both. These effects of CPA and TGFbeta1 were significantly more pronounced in GSTp(+) than in GSTp(-) hepatocytes. In conclusion, nuclear translocation of proTGFalpha increases in the course of hepatocarcinogenesis and appears to be involved in the inherent growth advantage of (pre)malignant hepatocytes. PMID- 15534612 TI - Preschool child care and risk of overweight in 6- to 12-year-old children. AB - OBJECTIVE: To determine if there is a relationship between center-based child care attendance from ages 3 to 5 y and future overweight at ages 6-12 y. DESIGN/METHODS: Longitudinal, observational study of child experience and future body mass index. SUBJECTS: A total of 1244 US children aged 6-12 y included in the 1997 Panel Study of Income Dynamics Child Development Supplement. MEASUREMENTS: Parent-reported child care attendance from ages 3 to 5 y, trichotomized as 'none', 'limited' (>0 but <15 h/week), and 'extensive' (> or =15 h/week). Overweight defined as a body mass index > or =95th percentile for age and gender. Candidate covariates (selected a priori): gender, race, age, poverty status, birth weight, hours of television per day, Behavior Problems Index score >90th percentile, and Home Observation for Measurement of the Environment-Short Form (HOME-SF) cognitive stimulation score. RESULTS: Of the potential confounding variables, race, HOME-SF cognitive stimulation score, and age significantly altered the relationship between child care attendance and overweight in the multiple logistic regression model. With these covariates in the final model, limited center-based child care attendance from ages 3 to 5 y was independently associated with a decreased risk of overweight at ages 6-12 y (adjusted odds ratio=0.56, 95% confidence interval 0.34, 0.93) relative to no child care attendance. Extensive center-based child care attendance was not associated with future overweight. CONCLUSIONS: Limited center-based child care attendance during the preschool years was independently associated with a decreased risk of future overweight relative to no child care attendance. Additional studies are needed to clarify these findings. PMID- 15534613 TI - Longitudinal changes in activity patterns, physical capacities, energy expenditure, and body composition in severely obese adolescents during a multidisciplinary weight-reduction program. AB - OBJECTIVE: To determine the longitudinal changes in body composition, physical capacities, and time and energy expenditure (EE) devoted to various activities in the course of a 9-month weight-reduction period. DESIGN: Longitudinal, clinical intervention including lifestyle education, moderate energy restriction, progressive training, and psychological follow-up. SUBJECTS: A total of 27 (13 boys and 14 girls) severely obese adolescents (mean BMI: 33.9 kg/m2; 41.5% fat mass (FM)), aged 12-16 y. MEASUREMENTS: Before the beginning and after the weight reduction program, body composition was assessed by dual-energy X-ray absorptiometry (DXA), physical capacities by multistage treadmill test, and EE both by whole-body calorimetry and in free-living conditions using the heart rate recording method. During 8 months of the weight-reduction period, type and duration of each activity were recorded using a daily controlled activity diary. RESULTS: One boy resigned after 5 months. Body weight (BW) and FM decreased (-19 and -42%, respectively, P<0.001) both in boys and in girls, but fat-free mass (FFM) decreased only in girls (-6%, P<0.001). VO2max (l/min) did not vary significantly, but strength and fitness were improved (P<0.001). Time and EE spent at sedentary activities decreased significantly (P<0.001) to the benefit of moderate (recreational) activities and total physical activities (P<0.001) at the institution during the weekdays, and at home during the weekends and holidays. CONCLUSION: The great BW and FM losses, preservation of FFM, and improvement of physical capacities of obese adolescents obtained under experimental conditions were associated with increases in leisure physical activities in free-living conditions at the expense of sleep and sedentary activities. PMID- 15534614 TI - Low physical activity levels of modern Homo sapiens among free-ranging mammals. AB - Obesity prevalence rates are increasing worldwide and one prevailing hypothesis is that physical activity levels of modern humans are markedly reduced compared to those of our Paleolithic ancestors. We examine this hypothesis by deriving relative activity energy expenditure from available doubly labeled water and indirect calorimetry data in free-ranging non-human mammals. Our results, given the constraints posed by limited data availability, suggest that a low physical activity level, much less than that observed in free-ranging non-human mammals or highly active humans, is present in modern adult humans living within advanced settings. Our observations lend support to the hypothesis that low activity related energy expenditure levels contribute to the rising worldwide prevalence of obesity. PMID- 15534615 TI - Physicians recommendations for and personal use of low-fat and low-carbohydrate diets. PMID- 15534616 TI - Dairy consumption and body mass index: an inverse relationship. AB - OBJECTIVE: To assess the relationship between consumption of dairy products and body mass index (BMI) in Tehranian adults. DESIGN: Cross-sectional study. SUBJECTS: A total of 462 healthy subjects (223 men and 239 women) aged over 16 y selected randomly from among participants of the Tehran Lipid and Glucose Study (TLGS). MEASUREMENTS: Dietary data were collected by means of a food frequency questionnaire for 1 y and two 24-h dietary recalls. Height and weight were measured and BMI was calculated. Physical activity was assessed by the Lipid Research Clinic (LRC) questionnaire. RESULTS: Consumption of dairy products was 3.7+/-1.0 and 2.9+/-1.2 servings per day in men and women, respectively. As the servings of dairy consumption increased per day, the proportion of normal-weight subjects rose and that of obese ones declined. As BMI increased, the proportion of subjects with lower consumption of dairy products increased, whereas that of those with higher consumption decreased. There was a significant inverse correlation between the servings of dairy consumption per day and BMI after controlling for the effect of age, physical activity, energy, carbohydrate, dietary fiber, protein and fat intake (r=-0.38, P<0.05). After adjustment for potential confounding variables, men and women in the top quartile of dairy consumption had lower chances for being overweight (OR=0.78, 95% CI=0.43-0.92 for men and OR=0.89, 95% CI=0.53-0.95 for women) and obese (OR=0.73, 95% CI=0.40-0.83 for men and OR=0.69, 95% CI=0.34-0.80 for women) compared to those in the first quartile. CONCLUSION: The results suggest an inverse relationship between dairy consumption and BMI. It is recommended that further studies address this issue by focusing on the dairy components responsible for this effect. PMID- 15534617 TI - Quantitative trait loci for abdominal fat and BMI in Hispanic-Americans and African-Americans: the IRAS Family study. AB - OBJECTIVE: To conduct linkage analysis for body mass index (BMI, kg/m2), waist-to hip ratio (WHR), visceral adipose tissue mass (VAT, cm2) and subcutaneous adipose tissue mass (SAT, cm2) using a whole genome scan. DESIGN: Cross-sectional family study. STUDY SUBJECTS: African-American families from Los Angeles (AA, n=21 extended pedigrees) and Hispanic-American families (HA) from San Antonio, TX (HA SA, n=33 extended pedigrees) and San Luis Valley, CO (HA-SLV, n=12 extended pedigrees), totaling 1049 individuals in the Insulin Resistance and Atherosclerosis (IRAS) Family Study. MEASUREMENTS: VAT and SAT were measured using a computed tomography scan obtained at the fourth and fifth lumbar vertebrae. All phenotypes were adjusted for age, gender, and study center. VAT, SAT, and WHR were analyzed both unadjusted and adjusted for BMI. RESULTS: Significant linkage to BMI was found at D3S2387 (LOD=3.67) in African-Americans, and at D17S1290 in Hispanic-Americans (LOD=2.76). BMI-adjusted WHR was linked to 12q13-21 (D12S297 (LOD=2.67) and D12S1052 (LOD=2.60)) in Hispanic-Americans. The peak LOD score for BMI-adjusted VAT was found at D11S2006 (2.36) in Hispanic families from San Antonio. BMI-adjusted SAT was linked to D5S820 in Hispanic families (LOD=2.64). Evidence supporting linkage of WHR at D11S2006, VAT at D17S1290, and SAT at D1S1609, D3S2387, and D6S1056 was dependent on BMI, such that the LOD scores became nonsignificant after adjustment of these phenotypes for BMI. CONCLUSIONS: Our findings both replicate previous linkage regions and suggest novel regions in the genome that may harbor quantitative trait locis contributing to variation in measures of adiposity. PMID- 15534618 TI - Netrin receptor deficient mice exhibit functional reorganization of dopaminergic systems and do not sensitize to amphetamine. AB - Netrins are guidance cues that play a fundamental role in organizing the developing brain. The netrin receptor, DCC (deleted in colorectal cancer), is highly expressed by dopaminergic (DA) neurons. DCC may therefore participate in the organization of DA circuitry during development and also influence DA function in the adult. Here we show that adult dcc heterozygous mice exhibit a blunted behavioral response to the indirect DA agonist amphetamine and do not develop sensitization to its effects when treated repeatedly. These behavioral alterations are associated with profound changes in DA function. In the medial prefrontal cortex, dcc heterozygotes exhibit increased tyrosine hydroxylase (TH) protein levels and dramatic increases in basal concentrations of DA and DA metabolites. In contrast, in the nucleus accumbens, dcc heterozygotes show no changes in either TH or DA levels, but exhibit decreased concentrations of DA metabolites, suggesting reduced DA activity. In addition, dcc heterozygous mice exhibit a small, but significant reduction in total number of TH-positive neurons in midbrain DA cell body regions. These results demonstrate for the first time that alterations in dcc expression lead to selective changes in DA function and, in turn, to differences in DA-related behaviors in adulthood. These findings raise the possibility that changes in dcc function early in life are implicated in the development of DA dysregulation observed in certain psychiatric disorders, such as schizophrenia, or following chronic use of drugs of abuse. PMID- 15534619 TI - Shared and specific susceptibility loci for schizophrenia and bipolar disorder: a dense genome scan in Eastern Quebec families. AB - The goal of this study was to identify susceptibility loci shared by schizophrenia (SZ) and bipolar disorder (BP), or specific to each. To this end, we performed a dense genome scan in a first sample of 21 multigenerational families of Eastern Quebec affected by SZ, BP or both (N=480 family members). This probably constitutes the first genome scan of SZ and BP that used the same ascertainment, statistical and molecular methods for the concurrent study of the two disorders. We genotyped 607 microsatellite markers of which 350 were spaced by 10 cM and 257 others were follow-up markers in positive regions at the 10 cM scan. Lander and Kruglyak thresholds were conservatively adjusted for multiple testings. We maximized the lod scores (mod score) over eight combinations (2 phenotype severity levels x 2 models of transmission x 2 analyses, affected/unaffected vs affected-only). We observed five genomewide significant linkages with mod score >4.0: three for BP (15q11.1, 16p12.3, 18q12-q21) and two for the shared phenotype, that is, the common locus (CL) phenotype (15q26,18q12 q21). Nine mod scores exceeded the suggestive threshold of 2.6: three for BP (3q21, 10p13, 12q23), three for SZ (6p22, 13q13, 18q21) and three for the CL phenotype (2q12.3, 13q14, 16p13). Mod scores >1.9 might represent confirmatory linkages of formerly reported genomewide significant findings such as our finding in 6p22.3 for SZ. Several regions appeared to be shared by SZ and BP. One linkage signal (15q26) appeared novel, whereas others overlapped formerly reported susceptibility regions. Despite the methodological limitations we raised, our data support the following trends: (i) results from several genome scans of SZ and BP in different populations tend to converge in specific genomic regions and (ii) some of these susceptibility regions may be shared by SZ and BP, whereas others may be specific to each. The present results support the relevance of investigating concurrently SZ and BP within the same study and have implications for the modelling of genetic effects. PMID- 15534620 TI - Use of bisphosphonates can dramatically improve pain in advanced hormone refractory prostate cancer patients. AB - INTRODUCTION: Approximately 85% of patients who die from prostate cancer present the spread of bone metastases. Even though the radiological appearance of such metastases is osteoblastic, it is now known that these lesions coexist in their microenvironment with blastic and lytic lesions. The process always begins with bone lysis by osteoclast proliferation, paralleling nearby bone deposition. The treatment options are palliative and have poor clinical response with short-lived improvement. We have studied the clinical effect of bisphosphonates (clodronate) in the treatment of skeletal complications from prostate cancer. MATERIALS AND METHODS: In an open prospective study, 58 patients with hormone-refractory prostate cancer with bone metastases were assessed from November 2000 to September 2003. The mean age was 70.3 y (range: 51-87 y). Bone scintigraphy, plain X-ray, assaying of prostate-specific antigen (PSA) and biochemical tests were requested before and following treatment. Patients were previously and subsequently assessed using the visual pain scale (0-10) and Karnofsky's index after the first and second intravenous (i.v.) infusions (administration of i.v. clodronate every 28 days) and every 4-6 months thereafter. Student's t-test was used for statistical analysis. RESULTS: A total of 53 patients (91.4%) showed improvement after the first and/or second cycle, which persisted for at least 4 months (average 6.3 months). The averages on the visual pain scale improved from 7.4 (range: 2-8) to 2.4 (0-7) and on Karnofsky's index from 43 (32-58) to 73 (50 82). The radiological appearance of the metastases improved in 27 patients (46.5%) and there were few relapses (six patients; 10.3%). CONCLUSIONS: Clodronate was effective in the treatment of skeletal complications from prostate cancer. There was an objective response in 91.4% of treated patients, with a marked improvement in the subjective visual pain scale evaluation as well as on Karnofsky's index, with low side effects. PMID- 15534621 TI - Long-term changes in the tibia and radius bone mineral density following spinal cord injury. AB - DESIGN: A prospective inception cohort study with an observational analytic design in a spinal cord injury (SCI) centre hospital. OBJECTIVE: To assess changes in trabecular and compact bone of the tibia and radius prospectively in subjects with SCI. SUBJECTS: In total, 10 individuals with an acute SCI. METHODS: Trabecular and compact bone density of the tibia and radius by peripheral quantitative computerised tomography. RESULTS: Analysis of the individual gradients of the curve coefficient showed changes in trabecular bone between 0.19 and -2.46 and in cortical bone between +0.07 and -0.93 in the tibia within 34 months after the SCI. Both trabecular and cortical bone showed a group mean loss of 99 mg/cm(3). No changes were observed in the radius. CONCLUSION: There is a major decrease in tibia mineral density over 3 years; however, no change is observed for the radius mineral content. Large interindividual differences existed in the patterns of loss in the tibia bone substance after SCI. These patterns indicate that there is no steady state of bone mineral density following 3 years of spinal cord injury. PMID- 15534622 TI - Participation and satisfaction after spinal cord injury: results of a vocational and leisure outcome study. AB - STUDY DESIGN: Survey. OBJECTIVES: Insight in (1) the changes in participation in vocational and leisure activities and (2) satisfaction with the current participation level of people with spinal cord injuries (SCIs) after reintegration in society. DESIGN: Descriptive analysis of data from a questionnaire. SETTING: Rehabilitation centre with special department for patients with SCIs, Groningen, The Netherlands. SUBJECTS: A total of 57 patients with traumatic SCI living in the community, who were admitted to the rehabilitation centre two to 12 years before the current assessment. MAIN OUTCOME MEASURES: Changes in participation in activities; current life satisfaction; support and unmet needs. RESULTS: Participation expressed in terms of hours spent on vocational and leisure activities changed to a great extent after the SCI. This was mainly determined by a large reduction of hours spent on paid work. While 60% of the respondents successfully reintegrated in work, many changes took place in the type and extent of the job. Loss of work was partially compensated with domestic and leisure activities. Sports activities were reduced substantially. The change in participation level and compensation for the lost working hours was not significantly associated with the level of SCI-specific health problems and disabilities. As was found in other studies, most respondents were satisfied with their lives. Determinants of a negative life satisfaction several years following SCI were not easily indicated. Reduced quality of life was particularly related to an unsatisfactory work and leisure situation. CONCLUSIONS: Most people with SCI in this study group were able to resume work and were satisfied with their work and leisure situation. PMID- 15534623 TI - High-dose methylprednisolone may cause myopathy in acute spinal cord injury patients. AB - STUDY DESIGN: Prospective cohort study. OBJECTIVE: Although Bracken et al have demonstrated a significant neuroprotective effect of high-dose intravenous (i.v.) methylprednisolone (MP) within 8 h post spinal cord injury (SCI), this practice has recently been challenged. We hypothesized it is possible that acute corticosteroid myopathy (ACM) may occur secondary to the MP. This pilot study was performed to test this hypothesis. SETTING: University of Miami School of Medicine/Jackson Memorial Hospital, Miami VA Medical Center, FL, USA. METHODS: Subjects included five nonpenetrating traumatic SCI patients, who received 24 h MP according to National Acute Spinal Cord Injury Studies (NASCIS) protocol, and three traumatic patients who suffered SCI and did not receive MP. Muscle biopsies and electromyography (EMG) were performed to determine if myopathic changes existed in these patients. RESULTS: Muscle biopsies from the SCI patients who received 24 h of MP showed muscle damage consistent with ACM in four out of five cases. EMG studies demonstrated myopathic changes in the MP-treated patients. In the three patients who had SCI but did not receive MP, muscle biopsies were normal and EMGs did not reveal evidence of myopathy. CONCLUSION: Our data suggest that MP in the dose recommended by the NASCIS may cause ACM. If this is true, part of the improvement of neurological recovery showed in NASCIS may be only a recording of the natural recovery of ACM, instead of any protection that MP offers to the injured spinal cord. PMID- 15534624 TI - Outcome of late surgical treatment in patients with incomplete paraplegia due to spinal degenerative diseases. AB - STUDY DESIGN: Retrospective analysis. OBJECTIVE: To assess the outcome of late surgical intervention in patients with incomplete paraplegia due to spinal degenerative diseases. SETTING: Three men and four women with cervical or thoracic spinal degenerative diseases, who preoperatively were unable to walk for more than 6 months in Mie prefecture, Japan. METHODS: Review of clinical records and questionnaire survey regarding the walking ability of patients 2 years after surgery. RESULTS: All seven patients were unable to walk postoperatively. CONCLUSION: A late surgical intervention may not lead to functional recovery in patients with spinal degenerative disease who were unable to walk for at least 6 months. PMID- 15534625 TI - CYP2A6 genetic polymorphisms and correlation with smoking status in Brazilians. AB - We investigated polymorphisms of cytochrome P450 2A6 (CYP2A6) and its association with smoking habits in 412 healthy Brazilians, self-recognized as white (n=147), black (n=123) and intermediate (n=142), and classified as smokers (n=205, including 61 ex-smokers) and nonsmokers (n=207). The frequencies of the variant alleles CYP2A6(*)1B, CYP2A6(*)2, CYP2A6(*)4 and CYP2A6(*)9 in the overall study population were 29.9, 1.7, 0.5 and 5.7%, respectively. Significant differences in the CYP2A6 allelic distribution were observed across the three population subgroups. There was a statistically significant trend for decreasing frequency of CYP2A6(*)1B from white to intermediate and to black persons. An association between CYP2A6 genotype and smoking dependence was detected, which could not be explained by the expected phenotypic activity of CYP2A6. In white and intermediate persons, the odds ratio (OR) of being smokers vs nonsmokers was 0.07 (95% CI 0.02-0.20; P<0.001) and 0.27 (95% CI 0.12-0.61; P<0.001), respectively, for genotypes including allele CYP2A6(*)1B, as compared to wild-type homozygous. In contrast, the corresponding OR in black Brazilians was 1.34 (95% CI 0.57-3.17; P=0.46). These data suggest that the CYP2A6(*)1B is associated with smoking dependence in white and intermediate, but not black Brazilians. PMID- 15534626 TI - Interethnic variability of ERCC2 polymorphisms. AB - Excision Repair Cross-Complementing Rodent Repair Group 2 (ERCC2) plays an important role in DNA repair by eliminating bulky DNA adducts produced by platinum agents during the nucleotide excision repair pathway. Several studies have associated polymorphisms in ERCC2 with response to platinum therapy, lung cancer risk, and DNA repair capacity. This study examined ERCC2 polymorphisms and haplotype structure across 18.9 kb in 95 European, 95 African, and 95 Asian individuals. Single-nucleotide polymorphisms (SNPs) (ERCC2 -9164 A>T, -1989 A>G, 516 G>A, 468 C>A [Arg156Arg], 1737 C>T [Val579Val], 2133 C>T [Asp711Asp], and 2251 T>G [Lys751Gln]) were mined and mapped using Golden Path, PolyMAPr, and Promolign. Genotyping was performed using PCR and pyrosequencing. Allele frequencies ranged from 0 to 0.47 (Europeans), 0.05 to 0.72 (Africans), and 0 to 0.47 (Asians). The synonymous cSNP at codon 579 could not be confirmed in our populations. There were significant differences in haplotype structure and frequency between populations. This information on ERCC2 genomic structure will allow the construction of definitive studies to clarify the clinical role of this important gene. PMID- 15534627 TI - [Dangerously tempting fellowships]. PMID- 15534628 TI - [When can generic preparations be used?]. PMID- 15534629 TI - [Occupational environment and health in oil industry]. PMID- 15534630 TI - [Sleep disorders in elderly patients who take hypnotics on a regular basis]. AB - BACKGROUND: Several studies have shown that the prevalence of sleep disorders such as sleep apnoea and periodic limb movements in sleep increases significantly with age. The assessment capacity is limited and there is a paucity of knowledge in this field among health professional, hence we hypothesise that these disorders are to a considerable extent under-diagnosed in the elderly, as the symptoms are often conceived of as insomnia. MATERIAL AND METHODS: Using ambulant clinical polysomnography, we investigated 19 patients aged 55 years or above who had been taking hypnotics on a daily basis (5 to 7 days a week) for at least one year. RESULTS: Among these 19 patients, the prevalence of sleep apnoea (AHI > 10) as well as periodic limb movements in sleep (PLM index > 10) was 42%; 74% had either sleep apnoea, periodic limb movements in sleep, or both. INTERPRETATION: Seen in relation to the fact that these elderly patients were given hypnotics for insomnia, the prevalence rates of sleep apnoea and periodic limb movements in sleep were high. We suggest that non-pharmacological alternatives should be considered and careful sleep assessment carried out before long-term prescription of hypnotics is instituted. PMID- 15534631 TI - [Physicians and drug industry: attitudes and practice]. AB - BACKGROUND: There is a need for more knowledge on how doctors assess the relationship between the pharmaceutical industry and the medical profession. METHOD: Postal questionnaires survey in 2002 among a representative sample of 1606 Norwegian doctors. The response rate was 73%. RESULTS: 70% of the respondents indicated that continued medical education would suffer without participation from the pharmaceutical industry, and 40 % that the industry was important for their own updating of skills. 52% agreed that doctors were influenced by the pharmaceutical industry, but 40% claimed that this was not the case for them personally. Specialists in internal medicine, surgery and family medicine were more likely to travel abroad sponsored by the pharmaceutical industry, whereas specialists in family medicine and internal medicine more often attended industry-sponsored meetings in Norway. Psychiatrists were more likely to see the relationship between doctors and industry as too regulated. Young doctors were more sceptical towards the pharmaceutical industry than their older colleagues. INTERPRETATION: Norwegian doctors find that the pharmaceutical industry is important to their own professional development and this may complicate attempts to regulate this relationship. It is, moreover, a problem that doctors probably underestimate the influence of the industry. PMID- 15534632 TI - [Cryobiology-- freeze preservation and storage of living cells and tissues]. AB - BACKGROUND: Freeze preservation and prolonged storage of living cells and tissue fragments have been met with increasing interest in medicine during the recent years, for diagnostic, therapeutic as well as research purposes. METHODS: The main methods: Programmed freezing, where a gradual decrease of temperature, usually 1 degrees C per minute is used, and vitrification: rapid freezing transforms cells and liquids to a solid state. In both cases chemical substances and macromolecules are added to reduce the risk of deleterious crystal formation in water. Such frozen cells can be stored for more than 10 to 15 years and retrieved in a viable condition. RESULTS AND INTERPRETATION: At present such techniques are used in reproductive medicine, with storage of human embryos after in vitro fertilization. In addition, there cells are increasing used for other therapeutic purposes, from haemopoietic stem cells to tissue fragments from skin and joint cartilage. Large biobanks of living, frozen cells are now being established in several countries. PMID- 15534633 TI - [Haemoglobin levels and transfusion practice among Norwegian anesthesiologists]. AB - BACKGROUND: Blood transfusions can be lifesaving, but definite transfusion triggers for different clinical situations need to be determined. MATERIAL AND METHODS: We carried out a survey among Norwegian anaesthesiologists regarding haemoglobin levels and transfusion triggers during surgery and in an intensive care setting. Blood conservation techniques were also explored. The results for surgical patients were compared with those from a similar survey in 1996. RESULTS: Compared with 1996, Norwegian anaesthesiologists now accept significantly lower transfusion triggers for surgical patients. Acceptable haemoglobin levels varied in the different patient examples in both the surgical and intensive care groups. With regard to surgery, junior doctors accept a lower transfusion trigger than do senior anaesthesiologists. A more liberal transfusion strategy is apparently used in patients with coronary disease. INTERPRETATION: Norwegian anaesthesiologists have changed their attitude to acceptable transfusion triggers over the 1996-2002 period. In an intensive care setting, they appear to have a more liberal transfusion policy compared to anaesthesiologists in other western European countries. PMID- 15534634 TI - [Lipid balance and cardiovascular risk in patients in a psychiatric long-term facility]. AB - Increase in weight and changes in lipid and glucose metabolism often occur in severe mental illness. It is established that this may hinge on the mental illness as such, and on lifestyle, resident living in institutions and on medication. We performed an explorative study of these factors in a general psychiatric long-term facility. MATERIAL: 30 resident patients gave fasting blood samples that were analysed for lipid and glucose variables. Body mass index, blood pressure and smoking habits were noted, as was medication history of neuroleptics. Patients were asked about family risk of coronary heart disease and diabetes. RESULTS: Body mass index was > 25 in 80% and > 30 in 40% of patients. Four patients had diabetes. Total cholesterol was > 6 mmol/l in 10 patients. The ratio of total cholesterol to HDL cholesterol was > 5 in 60%. An atherogenic index containing apolipoprotein measurements was above cut-off in 11 patients. The patients had experience with the use of a mean of 4.1 (SD = 2.1) different neuroleptics. 9 out of 12 patients who had used clozapin and/or olanzapin had a body mass index > 30. 25 patients smoked a mean of 17 cigarettes a day. INTERPRETATION: Most of our patients carry a high risk of coronary heart disease and diabetes. Even if this risk may partly be due to the mental illness itself, risk is also attributable to a combination of an unhealthy life style, medication and smoking. It is of utmost importance that health professionals, leisure time therapists and catering personnel do their best in order to reduce organisationally induced risk, as these patients cannot be expected to respond adequately to advice. PMID- 15534635 TI - [Side effects off allopurinol]. AB - Allopurinol is generally considered to be a safe and well tolerated drug. We report one patient with a serious effect from allopurinol, a serious exfoliative rash and signs of allopurinol hypersensitivity syndrome. Only after considerable diagnostic delay and patient morbidity, signs and symptoms were associated with the drug. The patient recovered when the medication was withdrawn. Data from the Norwegian reporting system for side effects of drugs for the period 1973-2003 show a wide range of side effects of allopurinol and even some fatalities. The indication for treatment must be carefully considered. Impaired renal function, ampicillin and thiazide diuretics increase the risk of serious hypersensitivity reactions. PMID- 15534637 TI - [Musculoskeletal disorders in the offshore oil industry]. AB - BACKGROUND: Musculoskeletal disorders are important causes of sick leave and disability among Norwegian offshore petroleum workers. More knowledge and interventions are needed in order to prevent this. In this review we consider prevalence and risk factors among offshore petroleum workers and point to the need for more research. METHOD: Literature searches on ISI Web of Science and PubMed were supplemented by reports from Norwegian offshore industry companies and the Norwegian Petroleum Directorate. RESULTS: Few studies were found on musculoskeletal disorders among offshore petroleum workers. The disorders are widespread, particularly among catering, construction and drilling personnel. It is not clear whether the prevalence is different from that among onshore workers. Risk factors are physical stressors and fast pace of work. Among catering personnel, these disorders are important causes of loss of the required health certificate but we could not identify any review of causes in the offshore industry generally. INTERPRETATION: More scientific studies are needed on musculoskeletal disorders as comparisons of prevalence and risk factors for offshore and onshore workers may point to more effective interventions. Better knowledge of the causes of loss of the health certificate may contribute to preventing early retirement. Interventions to prevent these disorders should be evaluated by controlled intervention studies. PMID- 15534636 TI - [Choice of proton pump inhibitors--does it play any role?]. PMID- 15534638 TI - [What do we know about chemical hazards in offshore work?]. AB - BACKGROUND: Norway has been an oil-producing nation for more than thirty years and a large number of Norwegians have been or are working on oil rigs. There are several chemical substances present on the oil platforms, and these factors may influence workers' health. MATERIAL AND METHODS: The international literature on offshore chemical exposure and health is summarised. RESULTS: The most important groups of chemical substances used on oil rigs are described: crude oil, production chemicals, asbestos and drilling chemicals. Different types of exposure during maintenance work are described as well. Very few exposure data are published. Acute, irritative health effects from chemical exposure are described, as well as chronic health effects like skin disorders and cancer. These workers seem to have a higher risk, that may be related to benzene exposure, of developing acute myelogenous leukemia. INTERPRETATION: Physicians who are treating patients working in the oil industry are advised to be aware of possible adverse health effects from the work environment on the rigs. Further exposure studies and research in this area are highly recommended, as the literature is scarce. PMID- 15534639 TI - [Return-to-work strategies in Statoil Forpleining]. AB - BACKGROUND: Since 1997, the catering department at Statoil, a Norwegian oil company, has worked systematically in order to facilitate employees' ability to avoid sick leaves and return to work after illness, and to improve their ability to cope with the challenges entailed in their work. MATERIAL AND METHODS: All employees in the catering department were sent a questionnaire about how they felt about their working environment. Sick leave data from Statoil and from Statistic Norway were collected. RESULTS: From 1997, sick leaves in catering have declined by 8%. 60% of employees and 90% of managers say that the physical as well as the psychosocial working environments have improved because of the programme described in this article. INTERPRETATION: Better support from co workers and supervisors has created a more positive culture in the workplace. PMID- 15534640 TI - [Patient-physician interaction over the internet]. AB - BACKGROUND: Nearly half of the Norwegian population claim that they would like to use the internet to communicate with their general practitioner. A web-based system complying with Norway's strict statutory requirements for the processing of personal data was developed and tested in an effort to assess the implications of this mode of communication. MATERIAL AND METHOD: The system was tested for one year in a group practice with six doctors. 200 patients were recruited and randomized into intervention and control groups. Data was collected through questionnaires, interviews and system logs. RESULTS: The 48 patients who used the system sent on average 3.3 messages, the six doctors sent between nine and 65 messages each. Traditional inquiries (visits, telephones) to the doctor averaged 3.2 and 4.5 for the intervention and control group respectively. 41% of the messages were inquiries about health issues, 22% were about renewals of prescriptions and sick leave notes, while 13% were requests for an appointment. Patients and doctors were both positive to this mode of communication. Patients who did not use the service said that they expected to use it in the future. INTERPRETATION: Electronic communication appears to replace some consultations and telephone inquiries. The study gives reason to expect that communication between patients and general practitioners over the internet will be more important in the future. PMID- 15534641 TI - [Challenges for professional development and research in general practice]. PMID- 15534642 TI - [Use of interdisciplinary experts groups in health technology assessment]. PMID- 15534643 TI - [Should the human smallpox virus (variola) be destroyed?]. AB - Smallpox, caused by variola virus, was a terror for civilizations around the world for more than 3000 years. Although the disease is eradicated, hundreds of variola virus isolates are kept in two WHO-collaborating facilities, one in USA and one in Russia. In spite of several agreements on destruction, it is now doubtful that these virus isolates will be destroyed. Variola virus may exist in other places and may be used as a biological weapon in war or for terror. Further research on variola virus is thus essential in order to achieve a better understanding of the pathogenicity of the virus and to develop new anti-variola virus vaccines and antiviral drugs. PMID- 15534646 TI - [The witness role of physicians]. PMID- 15534647 TI - [A group of physicians battling against]. PMID- 15534648 TI - [Physician for the patient--or a functionary of the system?]. PMID- 15534649 TI - [Research interpretation]. PMID- 15534650 TI - [Failing basis for the use of thiazides]. PMID- 15534652 TI - [Should I be treated with statins?]. PMID- 15534653 TI - [Children in facilities for asylum seeking persons--physician's role]. PMID- 15534654 TI - [Conflict of interest and therapy recommendations]. PMID- 15534656 TI - [Neonatal alloimmune thrombocytopenia]. PMID- 15534657 TI - [Atypical antipsychotics and dementia]. PMID- 15534659 TI - [Free access to information]. PMID- 15534660 TI - [Deliberate self-harm]. PMID- 15534661 TI - [Is top-level sport good for health?]. PMID- 15534662 TI - [Acute deliberate self-poisonings in the area of Trondheim, 1978-2002]. AB - BACKGROUND: Deliberate self-poisoning is a big health problem. We wanted to study if there had been changes in drug use, morbidity and mortality in this group over the last 25 years in our hospital's catchment area. MATERIAL AND METHODS: In this study, 924 patients admitted to our hospital after deliberate self-poisoning in 1978, 1987 and 2002 were studied prospectively. RESULTS: From 1978 to 1987, there was a significant increase in the incidence of self-poisoning followed by a decline from 1987 to 2002 among both men and women. The age distribution remained the same. Benzodiazepines were the most commonly used drugs during the whole period (20% of patients in 1978, 39% in 1987, and 30% in 2002). There has been a significant reduction in the use of acetylsalicylic acid, tricyclic antidepressants and a significant increase in paracetamol and selective serotonin reuptake inhibitor poisonings. The use of gastric lavage and activated charcoal declined. The main antidote in 1978 was physostigmine, in 1987 and 2002 n-acetyl cysteine. During the whole period, complications (usually minor) were recorded in approximately 10% of cases. In 1978, mortality was 1.3%, in 1987 0.9%; no patient died in 2002. INTERPRETATION: The incidence of deliberate self-poisoning has fallen over the last 25 years. Selective serotonin reuptake inhibitors have to a large extent replaced tricyclic antidepressants and paracetamol has replaced acetylsalicylic acid. PMID- 15534663 TI - [Surgical treatment of basal cell carcinoma]. AB - BACKGROUND: There are few prospective studies on the recurrence rates of basal cell carcinoma after different surgical modalities. MATERIAL AND METHODS: 263 primary basal cell carcinomas in as many patients were included from five dermatological departments in Norway. Patients on immunosuppressive medications were not included. All diagnoses were confirmed by histopathology. Primary end point was recurrence after five years, secondary end point cosmetic outcome. RESULTS: Observational data were available in 245 patients, in 160 at 5 years. Recurrence rate at 5 years was 7% after both surgical excision and cryosurgery. The cosmetic outcome was satisfactory in most cases. Tumour size was a risk factor for recurrence, localization was not. INTERPRETATION: Excision surgery and cryosurgery achieve low recurrence rates and acceptable cosmetic outcome in basal cell carcinoma. PMID- 15534664 TI - [Thought-provoking adaptation of otorhinolaryngologic surgery]. AB - BACKGROUND: Global budgeting was partially replaced by activity-based funding in 1999 in order to improve productivity and reduce waiting lists. The aim of this study is to estimate otolaryngologic surgery rates after the introduction of Diagnosis Related Groups funding. MATERIAL AND METHOD: National data on outpatient and inpatient otolaryngologic surgical procedures over the period 1999 to 2002 were provided by the Norwegian Patient Register; an estimate was made of the proportion undergoing uvuloplasty for sleep apnea or snoring. RESULTS: From 1999 to 2002, there was an overall increase in otolaryngologic procedures of 12%. Inter-procedural variation ranged from a 110% increase in uvuloplasties to a 12 % decrease in tonsillectomies. There was a particular increase in procedures requiring neither general anaesthesia nor a highly specialised surgeon. In spite of the increased rates of uvuloplasty, the proportion of patients selected for surgery remained unchanged. INTERPRETATION: The substantial increase in certain procedures may give rise to several interpretations: better medical technology, reallocation of surgical resources to disorders that had been inadequately covered, or a shift towards procedures for which marginal costs do not exceed treatment costs. PMID- 15534665 TI - [Hypertriglyceridemia--diagnostics, risk and treatment]. AB - BACKGROUND: Hypertriglyceridemia and low HDL cholesterol values are associated with adiposity, type 2 diabetes and metabolic syndrome. METHODS: This review article is based on literature studies, data from Rikshospitalet and Furst Medical Laboratory, sales figures for fibrates and clinical experience. RESULTS AND INTERPRETATION: Insulin resistance is a common contributory cause to hypertriglyceridaemia. In hypertriglyceridaemia, fasting glucose, HbA1c, TSH, creatinine, ALAT, ASAT, gamma GT, ALP and urine strips should be measured. The patient must be evaluated with regard to adiposity, use of alcohol, eating disorders and pregnancy. Direct measurement of LDL cholesterol gives a correct measure independent of the triglyceride level up to 13.0 mmol/L. LDL cholesterol levels may be low despite high total cholesterol levels and high triglyceride levels. Diet and lifestyle intervention is important. Blood sugar control is crucial. Statins are the first choice of drugs in combined hyperlipidaemia. Isolated hypertriglyceridaemia >10 mmol/L represents a large risk for pancreatitis. Purified omega-3 fatty acids, fibrates and eventually niacin are the drugs of choice in this condition. If LDL cholesterol levels are elevated, statins should also be considered. Combination of statin/fibrate or statin/niacin increase the risk of myopathy. PMID- 15534666 TI - [Arsenic poisoning]. AB - BACKGROUND: Arsenic poisoning has been discussed frequently in Norway during the past year on the background of a suspected crime case. There seem to be several uncertainties regarding this issue, also in the medical profession. MATERIAL AND METHODS: We have searched the literature and made a review based upon the present knowledge about arsenic and arsenic poisoning. RESULTS: Arsenic can be found in numerous chemical compounds with different properties. Inorganic arsenic compounds, like arsenic trioxide, are reactive and can cause damage to the body. Organic arsenic compounds, which are found in high concentrations in fish and shellfish, are not considered toxic. Ingestion of inorganic arsenic affects cellular energy production; lethal poisonings can occur. Common clinical features after acute intoxication with arsenic are dysphagia, nausea, vomiting, abdominal pain, diarrhoea, intense thirst, and muscle cramps. Clinical features of chronic poisoning are hyperkeratosis in the palms and foot soles, pigmentation and conjunctivitis. INTERPRETATION: Arsenic poisoning is a rare condition; its clinical features are uncharacteristic and the diagnosis must be confirmed by analysis of blood, urine and hair. PMID- 15534667 TI - [Epidural spinal cord stimulation in chronic refractory angina pectoris]. AB - BACKGROUND: Chronic refractory angina pectoris is defined as a condition with coronary insufficiency that cannot be controlled by a combination of medical therapy, angioplasty or bypass surgery. Different treatment options are evaluated in this patient group; spinal cord stimulation (SCS) is the one that is best documented. We have used this method since 1996 and present our experience. MATERIALS AND METHODS: From 1996 to 2001, spinal cord stimulators were implanted in 21 patients. A follow-up study was performed after 27 months (range 7-71) with a questionnaire. Preoperative clinical and angiographic data were retrieved from our records. RESULTS: 17 men and 4 women aged 55-88 years (median 73) were treated with SCS. Preoperatively all patients were in CCS (Canadian Cardiovascular Society) class III-IV. 81% had previously undergone coronary artery bypass surgery. At follow up, 81% were alive. Lead fracture occurred in two patients, displacement of the electrode in one. There were no other complications. 71% reported symptomatic improvement, mean CCS class was reduced from 3.5 prior to SCS to 2.2 at follow up, and use of nitroglycerin was reduced. INTERPRETATION: In chronic refractory angina pectoris, spinal cord stimulation is an alternative. We report "beginners' experience" that is positive including substantial symptomatic effect and a low complication rate. More hospitals in Norway should consider establishing SCS in order to offer this treatment option to a growing patient group. PMID- 15534668 TI - [Nine years' experience with endovascular treatment of abdominal aortic aneurysms]. AB - BACKGROUND: Endovascular repair of abdominal aortic aneurysm was started nine years ago at our institution. Our purpose was to evaluate the results over this period. MATERIAL AND METHODS: From 1995 to 2003, a total of 148 patients were treated for infrarenal abdominal aortic aneurysms by endovascular repair. The material includes 31 patients (21%) who were unfit for open surgery because of comorbidity or poor general condition. The patients have been followed up at regular intervals with clinical investigation, plain abdominal X-ray, and CT scans. Regional anaesthesia was used for 143 procedures; 5 were carried out under general anaesthesia. RESULTS: 30-day mortality following elective procedures was 2.3%. 31 endo-leaks were observed, 16 early and 15 late. Furthermore, 22 graft limb occlusions were diagnosed and thirteen of them repaired with femoro-femoral bypass. Secondary procedures were necessary in 42 (28.4%) of the patients, including 7 late conversions to open surgery. Some patients had more than one secondary procedure. With better stent grafts, the incidence of complications and secondary procedures has decreased significantly. During the last 4.5 years, only 4 secondary procedures (5.8%) have been necessary in 69 patients. In the total material the accumulated five-year survival rate was about 60%. INTERPRETATION: Results from endovascular repair of abdominal aortic aneurysm have improved in recent years. Ongoing prospective randomised studies could give us the information we need in order to establish what role this treatment should have. PMID- 15534669 TI - [Vacuum pack technique--a good method for temporal abdominal closure]. AB - Abdominal compartment syndrome(ACS) is a clinical entity characterised by increased intraabdominal pressure leading to multiple organ failure, fatal if left untreated. The treatment of abdominal compartment syndrome is surgical decompression with a temporary abdominal wall substitute. To avoid the development of abdominal compartment syndrome, temporary abdominal closure (TAC) should be considered after celiotomy for trauma. A new method for TAC was introduced at Ullevaal University Hospital in 2002, the "vac pac". METHOD: The "vac pac" technique is described and the patients treated with "vac pac" during the first year after introduction are presented. RESULTS: Five patients were treated using the "vac pac". One patient had acute pancreatitis and developed abdominal compartment syndrome. The other patients were severely injured and the indication for TAC was abdominal compartment syndrome in one patient, intestinal oedema in another, and damage control surgery with packing in two patients. Delayed primary closure was achieved within five days in all our patients. There were no fatalities and no complications related to the use of "vac pac" were registered. CONCLUSION: The "vac pac" technique seems to be a good method for TAC. PMID- 15534671 TI - [Kidney injuries and kidney failure caused by drugs]. PMID- 15534670 TI - [Results of surgical treatment of ventral hernia]. AB - BACKGROUND: The aim of this study was to examine the results of incisional hernia repair in our institution. MATERIAL AND METHODS: Repairs performed in from the start of 1998 to the end of 2001 were identified and the case notes reviewed. Some patients were also interviewed by telephone. Preoperative risk factors, type of repair, complications and recurrence were registered. RESULTS: 90 repairs were performed. The patients were followed up for 36 months on average. 58 (64%) patients had preoperative risk factors. 36 (40%) patients suffered complications, mostly seroma and wound infection. 39 (43%) patients experienced recurrence. DISCUSSION: Incisional hernia repair was associated with a high risk of complications and recurrence. PMID- 15534672 TI - [Are sleep difficulties in night work a problem for the offshore industry?]. AB - BACKGROUND: Shift work is increasingly used in industry and services; in the Norwegian offshore industry approximately 6750 employees work night shifts. We wanted to look into the implications of night shifts on sleep and circadian rhythm, with particularly focus on the offshore sector. MATERIAL AND METHODS: We conducted literature searches on Medline, covering the period from 1966 to 2003; only eight studies pertaining to the offshore activity in the North Sea with emphasis on sleep and night work were identified. RESULTS: Night shifts on oil platforms were associated with sleeping difficulties. The isolated setting on the installations in the North Sea probably facilitates biological adaptation to night work compared to work onshore. Light therapy has a documented effect for re adaptation to normal daytime functioning upon returning home. INTERPRETATION: Night work disturbs sleep. The level of functioning during the first night shifts is probably lower than the optimal level of functioning. Only a few studies of adaptation to night work in the offshore industry have been conducted and little research has been done concerning swing shifts in the North Sea. The field is obviously in need of more empirically based knowledge. PMID- 15534673 TI - [Shift work and accidents--relevance for the offshore industry]. AB - BACKGROUND: Increased use of shift work depends on keeping productivity and safety at acceptable levels. Approximately 7000 workers in the Norwegian offshore industry are working night shifts. MATERIAL AND METHODS: We have reviewed relevant literature on injuries in relation to shift work. RESULTS: The literature is inconclusive. The reason for this is most likely that all the studies are either observational or retrospective; no controlled interventions were found. However, shift work generally and night shifts in particular seem to increase the risk of injury. When newer data are taken into account, these clearly indicate that shift work is associated with reduced safety and productivity. INTERPRETATION: We conclude that if and when shift work is needed, it should be organised with certain patterns and rest breaks in order to reduce the risk of accidents and injuries. Focused and well-planned research is strongly needed in this field. Very few studies have been conducted on shift work and health effects offshore. PMID- 15534674 TI - [Can work-related eye injuries be avoided?]. AB - BACKGROUND: The eyes are among the organs most frequently hurt in occupational injuries. The characteristics of eye injuries were studied in order to suggest preventive measures. MATERIAL AND METHODS: Analyses were performed on work related eye injuries reported to the Norwegian Injury Surveillance System from a selection of emergency centres during the period 1990-2002, and on injuries reported by employers to the National Insurance Administration 1998-2001. RESULTS: The occurrence of injuries was stable over the period. Men sustained 94.4% of the injuries registered in injury surveillance system. The highest incidence was among those 20 to 24 years of age. Metal cutters and tools for polishing were involved in 25.7% of cases. Analyses of the injuries reported to the National Insurance Administration showed an odds ratio of 8.8 (95% CI 7.6 10.1) for injuries to workers in metal industry, 18.8 (95% CI 17.0-20.8) in automotive industry, and only 0.5 (95% CI 0.1-3.4) in oil refineries. Workers in oil refineries have a potentially hazardous work environment, but there, eye protection is mandatory. INTERPRETATION: There was no decline in the incidence of work-related eye injuries from 1990 to 2002. Workers in metal industry have a high risk of injuries and employers should consider requiring the use of eye protection. PMID- 15534675 TI - [Staffing and emergency service in Norwegian obstetrics and gynecology departments]. AB - BACKGROUND: We have made a survey of activities, skills, staffing and emergency services in Norwegian units for obstetrics and gynaecology. MATERIAL AND METHODS: A questionnaire was sent to all 43 units in February 2003. RESULTS: All units replied; 42 reported to have delivery service with emergency staffing. The reported annual number of births varied from 100 to 5600 (median 825, mean 1258). There was a large difference in the number of doctors on call in relation to annual number of births, with the larger units having a higher number of births per doctor on call. 60% of deliveries took place in units with a specialist always available. The numbers of doctors in each unit varied from 1 to 48 (median 8, mean 11). The outpatient activity was higher in local hospitals. Physicians with doctorates were mostly to be found in university hospitals. INTERPRETATION: The survey demonstrates a large variation in emergency services, distribution of posts and academic skills. This may reflect different functions and geographic conditions, though there are also large differences among comparable institutions. PMID- 15534676 TI - [Individual and group levels in population surveys and health services research]. PMID- 15534677 TI - [Injection centers--new international experiences and Norwegian political debate]. PMID- 15534679 TI - [New regulation fo autopsy]. PMID- 15534680 TI - [When the Parliament interfere with therapeutic methods]. PMID- 15534681 TI - [Between medicine, moral and moralism]. PMID- 15534682 TI - [Dietary supplements or not?]. PMID- 15534684 TI - [Overdiagnosis or wrong statement method?]. PMID- 15534685 TI - [Overdiagnosis and mammography screening]. PMID- 15534686 TI - [Chronic hepatitis C?]. PMID- 15534687 TI - [INTERHEART--a doubtful study]. PMID- 15534689 TI - [Is estrogen therapy of significance for the incidence of breast cancer?]. PMID- 15534691 TI - [Exhaustion--not fatigue]. PMID- 15534692 TI - Widespread A-to-I RNA editing of Alu-containing mRNAs in the human transcriptome. AB - RNA editing by adenosine deamination generates RNA and protein diversity through the posttranscriptional modification of single nucleotides in RNA sequences. Few mammalian A-to-I edited genes have been identified despite evidence that many more should exist. Here we identify intramolecular pairs of Alu elements as a major target for editing in the human transcriptome. An experimental demonstration in 43 genes was extended by a broader computational analysis of more than 100,000 human mRNAs. We find that 1,445 human mRNAs (1.4%) are subject to RNA editing at more than 14,500 sites, and our data further suggest that the vast majority of pre-mRNAs (greater than 85%) are targeted in introns by the editing machinery. The editing levels of Alu-containing mRNAs correlate with distance and homology between inverted repeats and vary in different tissues. Alu mediated RNA duplexes targeted by RNA editing are formed intramolecularly, whereas editing due to intermolecular base-pairing appears to be negligible. We present evidence that these editing events can lead to the posttranscriptional creation or elimination of splice signals affecting alternatively spliced Alu derived exons. The analysis suggests that modification of repetitive elements is a predominant activity for RNA editing with significant implications for cellular gene expression. PMID- 15534693 TI - Use of a dense single nucleotide polymorphism map for in silico mapping in the mouse. AB - Rapid expansion of available data, both phenotypic and genotypic, for multiple strains of mice has enabled the development of new methods to interrogate the mouse genome for functional genetic perturbations. In silico mapping provides an expedient way to associate the natural diversity of phenotypic traits with ancestrally inherited polymorphisms for the purpose of dissecting genetic traits. In mouse, the current single nucleotide polymorphism (SNP) data have lacked the density across the genome and coverage of enough strains to properly achieve this goal. To remedy this, 470,407 allele calls were produced for 10,990 evenly spaced SNP loci across 48 inbred mouse strains. Use of the SNP set with statistical models that considered unique patterns within blocks of three SNPs as an inferred haplotype could successfully map known single gene traits and a cloned quantitative trait gene. Application of this method to high-density lipoprotein and gallstone phenotypes reproduced previously characterized quantitative trait loci (QTL). The inferred haplotype data also facilitates the refinement of QTL regions such that candidate genes can be more easily identified and characterized as shown for adenylate cyclase 7. PMID- 15534695 TI - A historical study of structures for communication of organic chemistry information prior to 1950. AB - The evolution of the graphical structure diagram as a means of communication of chemical structure information is traced from its origins through to the mid-20th century. The impact of developments in structural theory on the representation of structures is discussed. A study of how structures were represented in the 19th and early 20th centuries in the Journal of the Chemical Society and its predecessors, the Journal of the American Chemical Society and United States patents was made, making use of electronic journal and patent archives. The problems associated with representing structures graphically are discussed. PMID- 15534696 TI - Representation and use of chemistry in the global electronic age. AB - We present an overview of the current state of public semantic chemistry and propose new approaches at a strategic and a detailed level. We show by example how a model for a Chemical Semantic Web can be constructed using machine processed data and information from journal articles. PMID- 15534694 TI - Conservation and evolution of cis-regulatory systems in ascomycete fungi. AB - Relatively little is known about the mechanisms through which gene expression regulation evolves. To investigate this, we systematically explored the conservation of regulatory networks in fungi by examining the cis-regulatory elements that govern the expression of coregulated genes. We first identified groups of coregulated Saccharomyces cerevisiae genes enriched for genes with known upstream or downstream cis-regulatory sequences. Reasoning that many of these gene groups are coregulated in related species as well, we performed similar analyses on orthologs of coregulated S. cerevisiae genes in 13 other ascomycete species. We find that many species-specific gene groups are enriched for the same flanking regulatory sequences as those found in the orthologous gene groups fromS. cerevisiae, indicating that those regulatory systems have been conserved in multiple ascomycete species. In addition to these clear cases of regulatory conservation, we find examples of cis-element evolution that suggest multiple modes of regulatory diversification, including alterations in transcription factor-binding specificity, incorporation of new gene targets into an existing regulatory system, and cooption of regulatory systems to control a different set of genes. We investigated one example in greater detail by measuring the in vitro activity of the S. cerevisiae transcription factor Rpn4p and its orthologs from Candida albicans and Neurospora crassa. Our results suggest that the DNA binding specificity of these proteins has coevolved with the sequences found upstream of the Rpn4p target genes and suggest that Rpn4p has a different function in N. crassa. PMID- 15534697 TI - Molecular similarity: a key technique in molecular informatics. AB - Molecular Informatics utilises many ideas and concepts to find relationships between molecules. The concept of similarity, where molecules may be grouped according to their biological effects or physicochemical properties has found extensive use in drug discovery. Some areas of particular interest have been in lead discovery and compound optimisation. For example, in designing libraries of compounds for lead generation, one approach is to design sets of compounds "similar" to known active compounds in the hope that alternative molecular structures are found that maintain the properties required while enhancing e.g. patentability, medicinal chemistry opportunities or even in achieving optimised pharmacokinetic profiles. Thus the practical importance of the concept of molecular similarity has grown dramatically in recent years. The predominant users are pharmaceutical companies, employing similarity methods in a wide range of applications e.g. virtual screening, estimation of absorption, distribution, metabolism, excretion and toxicity (ADME/Tox) and prediction of physicochemical properties (solubility, partitioning etc.). In this perspective, we discuss the representation of molecular structure (descriptors), methods of comparing structures and how these relate to measured properties. This leads to the concept of molecular similarity, its various definitions and uses and how these have evolved in recent years. Here, we wish to evaluate and in some cases challenge accepted views and uses of molecular similarity. Molecular similarity, as a paradigm, contains many implicit and explicit assumptions in particular with respect to the prediction of the binding and efficacy of molecules at biological receptors. The fundamental observation is that molecular similarity has a context which both defines and limits its use. The key issues of solvation effects, heterogeneity of binding sites and the fundamental problem of the form of similarity measure to use are addressed. PMID- 15534698 TI - BioSimGrid: towards a worldwide repository for biomolecular simulations. AB - BioSimGrid is a database for biomolecular simulations, or, a "Protein Data Bank extended in time" for molecular dynamics trajectories. We describe the implementation details: architecture, data schema, deposition, and analysis modules. We encourage the simulation community to explore BioSimGrid and work towards a common trajectory exchange format. PMID- 15534699 TI - Chemistry on the world-wide-web: a ten year experiment. AB - The server logs for access to the Cambridge Chemistry webserver show how use of the server has increased over the last ten years, with access doubling every year and a half. This growth has started to slow, and extrapolation of the data suggests that the current rate of access is close to a plateau of ten million downloads a year. The transition for chemists from no internet access to saturation coverage, therefore, appears almost complete. PMID- 15534700 TI - Enabling the exploration of biochemical pathways. AB - The Biochemical Pathways Wall Chart (http://www.expasy.org/tools/pathways/ref.1) has been converted into a molecule and reaction database. Major features of this database are that each molecule is represented by lists of all atoms and bonds (as connection tables), and in the reactions the reaction centre, the atoms and bonds directly involved in the bond rearrangement process, are marked. The information in the database has been enriched by a set of diverse 3D structure conformations generated by the programs CORINA and ROTATE. The web-based structure and reaction retrieval system C@ROL provides a wide range of search methods to mine this rich database. The database is accessible at http://www2.chemie.uni-erlangen.de/services/biopath/index.html and http://www.mol net.de/databases/biopath.html . PMID- 15534701 TI - Combined use of physicochemical data and small-molecule crystallographic contact propensities to predict interactions in protein binding sites. AB - Knowledge-based methods are a good alternative to force-field-based methods for the analysis of sites of interaction in protein binding cavities. Both the Protein Data Bank (PDB) and the Cambridge Structural Database (CSD) offer a good amount of data on non-covalent interactions. Although different from protein derived data, small-molecule crystal data from the CSD are worth looking at as they provide a much more abundant and diverse set of intermolecular contacts. CSD data, when properly corrected by use of octanol-water pi values, can be used to predict the type of ligand chemical group most likely to occupy a given position within a protein binding site. Comparison with observed positions of ligand groups shows that the success rates of these predictions vary from 23% to 84%. Often, the group predicted to be most preferred at a given position is similar but not identical to the observed ligand group; if these are considered successes, prediction success rates range from 71% to 94%. Using PDB data, the corresponding rates are 16% to 79%, and 61% to 96%. Specificity of prediction of NH groups is somewhat better when using PDB interaction data, but results of prediction of hydrophobic groups seem worse than those obtained with CSD data. We have analysed the importance of data selection by applying different filters to eliminate unwanted interactions from our knowledge-base. The presence of certain types of interactions can be undesirable if they are unrepresentative of biological situations (contact to solvent molecules in small-molecule crystal structures, secondary crystallographic contacts) or if they are likely to add noise to the data without conveying much new information (long-distance contacts, sparsely-populated data sets). The elimination of solvent contacts was found to have no effect on the prediction of ligand groups in our test set. Both secondary contact filtering and noise filtering were found to have a clear beneficial effect on predictive ability. PMID- 15534702 TI - A structure-odour relationship study using EVA descriptors and hierarchical clustering. AB - Structure-odour relationship analyses using hierarchical clustering were carried out on a diverse dataset of 47 molecules. These molecules were divided into seven odour categories: ambergris, bitter almond, camphoraceous, rose, jasmine, muguet, and musk. The alignment-independent descriptor EVA (EigenVAlue) was used as the molecular descriptor. The results were compared with those of another kind of descriptor, the UNITY 2D fingerprint. The dendrograms obtained with these descriptors were compared with the seven odour categories using the adjusted Rand index. The dendrograms produced by EVA consistently outperformed those from UNITY 2D in reproducing the experimental odour classifications of these 47 molecules. PMID- 15534703 TI - Comparison of topological descriptors for similarity-based virtual screening using multiple bioactive reference structures. AB - This paper reports a detailed comparison of a range of different types of 2D fingerprints when used for similarity-based virtual screening with multiple reference structures. Experiments with the MDL Drug Data Report database demonstrate the effectiveness of fingerprints that encode circular substructure descriptors generated using the Morgan algorithm. These fingerprints are notably more effective than fingerprints based on a fragment dictionary, on hashing and on topological pharmacophores. The combination of these fingerprints with data fusion based on similarity scores provides both an effective and an efficient approach to virtual screening in lead-discovery programmes. PMID- 15534704 TI - Predicting protein-ligand binding affinities: a low scoring game? AB - We have investigated the performance of five well known scoring functions in predicting the binding affinities of a diverse set of 205 protein-ligand complexes with known experimental binding constants, and also on subsets of mutually similar complexes. We have found that the overall performance of the scoring functions on the diverse set is disappointing, with none of the functions achieving r(2) values above 0.32 on the whole dataset. Performance on the subsets was mixed, with four of the five functions predicting fairly well the binding affinities of 35 proteinases, but none of the functions producing any useful correlation on a set of 38 aspartic proteinases. We consider two algorithms for producing consensus scoring functions, one based on a linear combination of scores from the five individual functions and the other on averaging the rankings produced by the five functions. We find that both algorithms produce consensus functions that generally perform slightly better than the best individual scoring function on a given dataset. PMID- 15534705 TI - New horizons in mouse immunoinformatics: reliable in silico prediction of mouse class I histocompatibility major complex peptide binding affinity. AB - Quantitative structure-activity relationship (QSAR) analysis is a main cornerstone of modern informatic disciplines. Predictive computational models, based on QSAR technology, of peptide-major histocompatibility complex (MHC) binding affinity have now become a vital component of modern day computational immunovaccinology. Historically, such approaches have been built around semi qualitative, classification methods, but these are now giving way to quantitative regression methods. The additive method, an established immunoinformatics technique for the quantitative prediction of peptide-protein affinity, was used here to identify the sequence dependence of peptide binding specificity for three mouse class I MHC alleles: H2-D(b), H2-K(b) and H2-K(k). As we show, in terms of reliability the resulting models represent a significant advance on existing methods. They can be used for the accurate prediction of T-cell epitopes and are freely available online ( http://www.jenner.ac.uk/MHCPred). PMID- 15534706 TI - The semantic smart laboratory: a system for supporting the chemical eScientist. AB - One goal of eScience is to enable the end-to-end publication of experiments and results. In the Combechem project we have developed an innovative human-centred system which captures the process of a chemistry experiment from plan to execution. The system comprises an electronic lab book replacement, which has been successfully trialled in a synthetic organic chemistry laboratory, and a flexible back-end storage system. Working closely with the users, we found that a light touch and a high degree of flexibility was required in the user interface. In this paper, we concentrate on the representation and storage of human-scale experiment metadata, introducing an ontology to describe the record of an experiment, and a storage system for the data from our lab book software. Just as the interfaces need to be flexible to cope with whatever a chemist wishes to record, so the back end solutions need to be similarly flexible to store any metadata that may be created. The storage system is based on Semantic Web technologies, such as RDF, and Web Services. It gives a much higher degree of flexibility to the type of metadata it can store, compared to the use of rigid relational databases. PMID- 15534707 TI - Chemical documents: machine understanding and automated information extraction. AB - Automatically extracting chemical information from documents is a challenging task, but an essential one for dealing with the vast quantity of data that is available. The task is least difficult for structured documents, such as chemistry department web pages or the output of computational chemistry programs, but requires increasingly sophisticated approaches for less structured documents, such as chemical papers. The identification of key units of information, such as chemical names, makes the extraction of useful information from unstructured documents possible. PMID- 15534708 TI - Optimising the EVA descriptor for prediction of biological activity. AB - EVA is a multivariate molecular descriptor for use in QSAR studies. It is constructed from vibrational eigenvalues derived from either a quantum theoretical or molecular mechanical treatment of molecular structure. This paper applies the method to biological-activity data using measures of the inotropic potential of a range of Calcium channel agonists. The performance of the descriptor, as both an explanatory and a predictive tool, is analysed in relation to the way in which it is constructed using a rigorous statistical treatment. Its capabilities are examined in relation to those of previously published methodology which used a composite descriptor. It is shown to have improved performance and several procedural advantages, such as ease of calculation and operation. It is a 3-D structural descriptor which does not require prior co alignment of structures for a QSAR study. PMID- 15534709 TI - Meerwein-Ponndorf-Verley alkynylation of aldehydes: essential modification of aluminium alkoxides for rate acceleration and asymmetric synthesis. AB - A novel carbonyl alkynylation has been accomplished based on utilization of the Meerwein-Ponndorf-Verley (MPV) reaction system. The success of the MPV alkynylation crucially depends on the discovery of the remarkable ligand acceleration effect of 2,2'-biphenol. For example, the alkynylation of chloral (2c) with the aluminium alkoxide 6(R = Ph), prepared in situ from Me(3)Al, 2,2' biphenol and 2-methyl-4-phenyl-3-butyn-2-ol (1a) as an alkynyl source, proceeded smoothly in CH(2)Cl(2) at room temperature to give the desired propargyl alcohol 3ca in almost quantitative yield after 5 h stirring. The characteristic feature of this new transformation involving no metal alkynides can be visualized by the fact that the alkynyl group bearing keto carbonyl was transferred successfully to aldehyde carbonyl without any side reactions on keto carbonyl. Although the use of (S)-1,1[prime or minute]-bi-2-naphthol and its simple analogues was found to be unsuitable for inducing asymmetry in this reaction, design of new chiral biphenols bearing a certain flexibility of the biphenyl axis led to satisfactory results in terms of enantioselectivity as well as reactivity. PMID- 15534710 TI - Differentially-protected steroidal triamines; scaffolds with potential for medicinal, supramolecular, and combinatorial chemistry. AB - Cholic acid 2a has been converted into two new orthogonally-protected triamino scaffolds, 13 and 14. The synthesis proceeds via the bis-Boc-NH-substituted azide 10, for which an improved preparation is described. After removal of the Boc groups, the two axial amines are differentiated through a novel monoprotection employing 1-(2-nitrobenzenesulfonyloxy)-benzotriazole 29. Regioselectivity of > or 50 : 1 is achieved, presumably reflecting an exceptional sensitivity to steric hindrance. Protection of the remaining amino group as Boc or Alloc gives the scaffolds in approximately 40% overall yield from cholic acid. Scaffold 13 has been sequentially deprotected and derivatised with N-carbamoyl amino acids, to give a model for tripodal peptide libraries. PMID- 15534711 TI - Synthesis and stability of small molecule probes for Pseudomonas aeruginosa quorum sensing modulation. AB - The human pathogen Pseudomonas aeruginosa uses N-butyryl-L-homoserine lactone (BHL) and N-(3-oxododecanyl)-L-homoserine lactone (OdDHL) as small molecule intercellular signals in a phenomenon known as quorum sensing (QS). QS modulators are effective at attenuating P. aeruginosa virulence; therefore, they are a potential new class of antibacterial agent. The lactone in BHL and OdDHL is hydrolysed under physiological conditions. The hydrolysis proceeds at a rate faster than racemisation of the alpha-chiral centre. Non-hydrolysable, non racemic analogues (small molecule probes) were designed and synthesised, replacing the lactone with a ketone. OdDHL analogues were found to be relatively unstable to decomposition unless they were difluorinated between the beta-keto amide. Stability studies on a non-hydrolysable, cyclohexanone analogue indicated that racemisation of the alpha-chiral centre was relatively slow. This analogue was assayed to show that the L-isomer is likely to be responsible for the QS autoinducing activity in P. aeruginosa and Serratia strain ATCC39006. PMID- 15534712 TI - Kinetic resolution of tert-butyl (RS)-3-alkylcyclopentene-1-carboxylates for the synthesis of homochiral 3-alkyl-cispentacin and 3-alkyl-transpentacin derivatives. AB - High levels of stereocontrol are observed in the conjugate addition of lithium dibenzylamide to tert-butyl (RS)-3-alkylcyclopentene-1-carboxylates (alkyl = Et, Bn), with addition occurring exclusively anti- to the 3-alkyl substituent. Treatment of a range of tert-butyl (RS)-3-alkylcyclopentene-1-carboxylates (alkyl = Et, Bn, (i)Pr, (t)Bu) with lithium (RS)-N-benzyl-N-[small alpha] methylbenzylamide indicates that good enantiorecognition is observed (E > 80) in their mutual kinetic resolution. In these reactions, conjugate addition of the lithium amide occurs exclusively anti- to the 3-alkyl substituent, with subsequent C(1)-protonation occurring preferably anti- to the 2-amino group in the 3-Et, 3-Bn and 3-(i)Pr cases, giving predominantly the corresponding 1,2-syn 2,3-anti-diastereoisomers. Conjugate addition to (RS)-3-tert-butyl cyclopentene-1 carboxylate results in exclusive 2,3-anti -addition and a reversal in C(1) protonation selectivity, giving predominantly the 1,2-anti-2,3-anti diastereoisomer. Furthermore, the kinetic resolution of the tert-butyl (RS)-3 alkylcyclopentene-1-carboxylates (alkyl = Et, Bn, (i)Pr, (t)Bu) with lithium (S) N-benzyl-N-alpha-methylbenzylamide proceeds efficiently, giving, at between 47 and 51% conversion, the resolved 3-alkylcyclopentene-1-carboxylates in >85 to >98% ee and the beta-amino ester products of conjugate addition in high de, consistent with E > 80 in each case. Subsequent deprotection of the 1,2-syn-2,3 anti-3-alkyl-beta-amino esters (alkyl = Et, Bn, (i)Pr) by hydrogenolysis and ester hydrolysis gives the corresponding 1,2-syn-2,3-anti-3-alkylcispentacins in >98% de and 98 +/- 1% ee. Selective epimerisation of the 1,2-syn-2,3-anti-3-alkyl beta-amino esters (alkyl = Et, Bn, (i)Pr, (t)Bu) by treatment with KO(t)Bu in (t)BuOH gives the corresponding 1,2-anti-2,3-anti-3-alkyl-beta-amino esters in quantitative yield and in >98% de, with subsequent deprotection by hydrogenolysis and ester hydrolysis giving the corresponding 1,2-anti-2,3-anti-3 alkylcispentacin hydrochlorides in >98% de. PMID- 15534713 TI - Parallel kinetic resolution of tert-butyl (RS)-3-alkyl-cyclopentene-1 carboxylates for the asymmetric synthesis of 3-alkyl-cispentacin derivatives. AB - The double mutual kinetic resolution of tert-butyl (RS)-3-benzyl-cyclopentene-1 carboxylate with a 50 : 50 mixture of lithium (RS)-N-benzyl-N-alpha methylbenzylamide and lithium (RS)-N-3,4-dimethoxybenzyl-N-alpha methylbenzylamide gives, after protonation with 2,6-di-tert-butylphenol, a 50 : 50 mixture of the readily separable N-benzyl-(1SR,2RS,3RS,alphaRS)- and N-3,4 dimethoxybenzyl-(1SR,2RS,3RS,alphaRS)-beta-amino esters in >98% de in each case. This product distribution indicates that these amides react at very similar rates and with no mutual interference to furnish readily separable products, and are thus ideal for parallel kinetic resolution. The efficient parallel kinetic resolution (E > 65) of a range of tert-butyl (RS)-3-alkyl-cyclopentene-1 carboxylates with a pseudoenantiomeric mixture of homochiral lithium (S)-N-benzyl N-alpha-methylbenzylamide and lithium (R)-N-3,4-dimethoxybenzyl-N-alpha methylbenzylamide gives, after separation and N-deprotection, a range of carboxylate protected 3-alkyl-cispentacin derivatives in >98% de and >95% ee. PMID- 15534714 TI - Ethynyl pi-extended 2,5-diphenyl-1,3,4-oxadiazoles and 2-phenyl 5-(2-thienyl) 1,3,4-oxadiazoles: synthesis, X-ray crystal structures and optical properties. AB - 2-(4-tert-Butylphenyl)-5-(4-ethynylphenyl)-1,3,4-oxadiazole reacts with a series of heteroaryl iodides under standard Sonogashira cross-coupling conditions (Pd[PPh(3)](2)Cl(2), CuI, triethylamine, THF) to yield products 2a-g in 40-79% yields (heteroaryl = 2-pyridyl, 3-pyridyl, 4-pyridyl, 2-pyrazyl, 5-bromo-2 pyrimidyl, 2-thienyl and 3-thienyl, respectively). Compound 2f was lithiated followed by electrophilic iodination (BuLi, perfluorohexyl iodide) to give 3, which by a two-step sequence gave the terminal ethynylthienyl derivative 5. Conversion of 5 into the terminal ethynylaldehyde derivative 7, via acetal derivative 6, proceeded in high yield. Starting from 2-iodo-5 methoxycarbonylthiophene, a five-step sequence afforded 2-(4-tert-butylphenyl)-5 (4-ethynylthienyl)-1,3,4-oxadiazole 13 (13% overall yield). Reactions of 13 gave terminal pyridyl, pyrazyl, pyrimidyl and thienyl derivatives, analogous to those obtained from 1. Two-fold reaction of 13 with 2,5-diiodothiophene gave the bis(ethynylthienyl)thiophene derivative 15 (30% yield). Solution UV-Vis absorption and photoluminescence spectra establish that replacement of the phenyl ring in the 2,5-diphenyl-1,3,4-oxadiazole series 2a-g by a thienyl ring [i.e. the 2-phenyl-5-(2-thienyl)-1,3,4-oxadiazole series 14a-g] leads to a red shift in the lowest energy band in both the absorption spectra and emission spectra. The X-ray crystal structures of compounds 2d, 2g, 5 and 14d.CHCl(3) reveal that the molecular structures are approximately planar although there are substantial differences in the conformations. PMID- 15534715 TI - Synthesis of ring-oxidized retinoids as substrates of mouse class I alcohol dehydrogenase (ADH1). AB - Ring-oxidized retinoids have been synthesized stereoselectively using the Stille cross-coupling reaction. Kinetic constants of mouse class I alcohol dehydrogenase (ADH1) with these retinoids were determined. PMID- 15534716 TI - Enantioselective synthetic thalidomide receptors based upon DNA binding motifs. AB - A series of molecularly imprinted polymer (MIP) synthetic receptors selective for the sedative thalidomide (5) have been designed and synthesized based upon the functional monomer 9-(2'-methacryloyloxyethyl)adenine (2). (1)H-NMR studies were used to establish the existence of DNA-like binding interactions between 2 and the template (5). A series of ethylene glycol dimethacrylate cross-linked copolymers was synthesized using either 2 or methacrylic acid, or a combination of these functional monomers. Zonal HPLC studies demonstrated enantioselectivity (alpha = 2.11) and ligand selectivity which could be attributed to the interaction of 2 with the imide moiety of 5. Compound 2 provided a more significant contribution to the binding of 5 than methacrylic acid, though a combination of these two functional monomers resulted in improved enantioselectivity. Frontal chromatographic and batch binding studies confirmed the observed differences in affinity of the imprinted and reference polymers for the template. PMID- 15534717 TI - Sequential hydroformylation/aldol reactions: versatile and controllable access to functionalised carbocycles from unsaturated carbonyl compounds. AB - Three different modes of hydroformylation/aldol reaction sequences involving either acid-catalysed aldol reactions, Mukaiyama aldol addition of pre-formed enolsilanes or aldol addition of in situ generated boron enolates can be applied to unsaturated ketones and ketoesters to afford the corresponding carbocyclic aldol adducts in good yields proceeding through the intermediate activated ketoaldehydes. In selected cases, complimentary, synthetically useful diastereoselectivities were observed in the products. PMID- 15534718 TI - Double diastereoselective SuperQuat glycolate aldol reactions: application to the asymmetric synthesis of polyfunctionalised lactones. AB - Polyfunctionalised lactones with up to five contiguous stereocentres may be prepared with high stereocontrol by a double diastereoselective aldol protocol with protected homochiral alpha,beta-dihydroxy- or alpha,beta-gamma trihydroxyaldehydes and a chiral glycolate oxazolidinone, followed by subsequent O-desilylation and lactonisation. PMID- 15534719 TI - Enantioselective catalysis in water: Mukaiyama-aldol condensation promoted by copper complexes of bisoxazolines supported on poly(ethylene glycol). AB - (S)-3-Phenyl-2-aminopropanol-derived bisoxazolines supported on a modified poly(ethylene glycol) were shown to be effective Cu(II) ligands for the enantioselective Mukaiyama-aldol condensation of various aldehydes with the trimethylsilyl keteneacetal of methyl isobutyrate carried out in water. Enantiomeric excesses comparable to those obtained with nonsupported ligands in the same solvent were observed. The solubility of the ligand in water, ensured by the presence of the polymeric support, allowed a very convenient catalyst recycling procedure involving simple removal of the reaction product by extraction in Et(2)O and addition of fresh reagents to the catalyst-containing aqueous solution. The chemical and stereochemical efficiency of the catalyst was only marginally eroded over its use in three reaction cycles. PMID- 15534720 TI - Beta-blocker therapy failures in symptomatic probands with genotyped long-QT syndrome. AB - Beta-blocker therapy is one of the principal therapies for congenital long-QT syndrome (LQTS). However, breakthrough cardiac events occur while being treated with beta-blockers. We sought to determine the frequency of and clinical correlates underlying beta-blocker therapy failures in genotyped, symptomatic LQTS probands. The medical records were analyzed only for genotyped LQTS probands who presented with a LQTS-attributable clinical event and were receiving beta blocker therapy. The study cohort comprised 28 such patients: 18 KCNQ1/KVLQT1(LQT1), 7 KCNH2/HERG (LQT2), and 3 SCN5A (LQT3). The prescribed beta blocker was atenolol (12), propranolol (10), metoprolol (4), and nadolol (2). Beta-blocker therapy failure was defined as breakthrough cardiac events including syncope, aborted cardiac arrest (ACA), appropriate implantable cardioverter defibrillator (ICD) therapy, or sudden death occurring while on beta-blocker therapy. During a median follow-up of 46 months, 7/28 (25%) LQTS probands experienced a total of 15 breakthrough cardiac events. Their initial presentation was ACA (3), bradycardia during infancy (2), and syncope (2). The underlying genotype was KVLQT1 (6) and HERG (1). Two breakthroughs were attributed to noncompliance. Of the 13 breakthroughs occurring while compliant, 10 occurred with atenolol and 3 with propranolol (p = 0.03). In this study cohort, one-fourth of genotyped LQTS probands failed beta-blocker therapy. Treatment with atenolol, young age at diagnosis, initial presentation with ACA, KVLQT1 genotype, and noncompliance may be important factors underlying beta-blocker therapy failures. PMID- 15534721 TI - Systematic-to-pulmonary collaterals: a source of flow energy loss in Fontan physiology. AB - Patients with Fontan-modified, single-ventricle heart frequently have systemic collaterals that increase pulmonary blood flow. Competitive flow from these auxiliary vessels can also elevate pulmonary artery pressure, a process leading to erosion of flow's mechanical energy. An analogous analytical description of mixing fluid streams was used to provide insight into flow energetics associated with systemic-to-pulmonary collaterals in Fontan-type circulation. We find that theoretical pressure increases and flow energy losses due to mixing vary quadratically as the velocity differences of the interacting fluid streams. Moreover, the predicted flow energy loss is shown to depend directly on the resultant pressure increase. Based on studies of aortopulmonary collaterals in patients with Fontan anatomy, we provide an estimate of pulmonary artery pressure elevation and flow energy loss, factors that are of considerable clinical importance. PMID- 15534722 TI - Paroxysmal atrioventricular block in young patients. AB - We describe 17 patients (8 girls, and 9 boys), aged 9.6 +/- 5.7 years, with paroxysmal atrioventricular block (PAVB), a condition rarely described in children. Holter monitoring documented the PAVB in 15 patients, and tilt test was performed in 4 patients (positive in 1). The electrocardiograph (ECG) was normal in 7 patients. Two patients had acquired and 11 patients had congenital heart disease (CHD). Syncope or presyncope were present in 7 patients. A normal ECG was significantly more frequent in symptomatic patients. Pauses were significantly longer in girls and in children <5 years. PAVB was recorded only during nocturnal hours in 6 patients and throughout the day in the others. The sinus rate decreased during PAVB in 6 patients and increased in 4 (generally younger girls with symptoms). Permanent pacemakers were implanted in 13 patients, including 7 asymptomatic patients with CHD and severe bradycardia. During follow-up (3.7 +/- 2.5 years), 1 patient developed complete AVB. Although PAVB was still present in 91% of paced patients, symptoms did not recur because pacing prevented the pauses. In conclusion, PAVB is a rare arrhythmia. Autonomic nervous system dysfunction seems to play an etiological role and permanent pacing was an effective treatment. PMID- 15534723 TI - High energy external cardioversion for refractory atrial fibrillation in postoperative tetralogy of fallot. AB - Long-term complications of surgical repair of Tetralogy of Fallot include atrial arrhythmias. These can be difficult to treat, and loss of sinus rhythm can lead to profound hemodynamic consequences in the presence of residual structural abnormalities. We describe the first report of high-energy external cardioversion in a 46-year-old man with repaired tetralogy of Fallot with atrial fibrillation refractory to conversion with normal energy. This represents an alternative to internal cardioversion or rate control for these patients. PMID- 15534724 TI - Amplatzer closure of atrial septal defect and da Vinci robot-assisted repair of vascular ring. AB - Technology for minimally invasive approaches to congenital heart disease is a rapidly evolving field. This case report reviews a novel approach to combining two of the newer technologies available to treat a pediatric patient with an atrial septal defect (ASD) and a vascular ring. This report is the first to describe the use of the da Vinci surgical system to assist in a thoracoscopic procedure for a pediatric patient. The da Vinci assisted division of the vascular ring, joined with an Amplatzer closure of the ASD, demonstrates how maximum benefit can be obtained for patients by combining emerging technologies. PMID- 15534725 TI - Aspirin treatment for neonatal infectious endocarditis. AB - We describe a term female neonate with Serratia marcescens endocarditis. Despite adequate antibiotic therapy for 8 days, the bacteremia persisted and there was an increase in vegetation size. Treatment with aspirin was initiated, with resolution of the bacteremia and a gradual decrease in vegetation size. We conclude that in neonatal endocarditis, aspirin may be beneficial additional treatment. PMID- 15534726 TI - Anomalous origin of pulmonary artery from the innominate artery. PMID- 15534727 TI - The effect of basic and advanced pediatric life support maneuvers. PMID- 15534728 TI - Descending thoracic and thoracoabdominal aortic aneurysm in patients with Takayasu's disease. AB - From June 1974 to December 2001 we performed operative treatment on 33 patients with descending thoracic or thoracoabdominal aortic aneurysm in association with Takayasu disease. There were 25 men and 8 women with a mean age of 40.2A years (range 16-64A years). Nineteen patients came from North Africa, 6 were from France, and 8 were from various locations in the world. The revealing symptom was hypertension in 12 cases, thoracic or abdominal pain in 7, isolated inflammatory syndrome in 5, neurologic or ocular manifestations in 3, rupture in 3, and embolization to the lower extremity in 1. In the remaining two cases discovery was coincidental. The aneurysm was confined to the thoracic aorta in 10 cases and involved both the thoracic and abdominal aorta in 23 cases. There were 8 type I, 6 type II, 4 type III, and 5 type IV aneurysms according to Crawford's classification. Two patients had undergone previous repair of the thoracoabdominal aorta. Four patients required first-stage treatment of a renal artery lesion to control hypertension. Six patients had associated aneurysms of the proximal aorta, including five treated via the distal elephant trunk technique in first-stage procedures. Aneurysm repair consisted of prosthetic replacement of the thoracoabdominal aorta in 31 cases, exclusion bypass in 1 case, and stent graft placement in 1 case. The procedure was performed with cross clamping alone in 13 cases, distal perfusion in 17 cases, and deep hypothermic circulatory arrest in 3 cases. Twenty patients (61%) had associated renal and/or intestinal artery lesions that were treated during the same procedure as that for the thoracoabdominal aorta in 19 patients (58%). A total of 24 procedures were performed on renal arteries (17 revascularizations, 7 nephrectomies). Associated supraaortic trunks lesions were present in 15 patients (45%) and were treated in 12 patients, including 8 in first-stage procedures prior to thoracoabdominal aortic aneurysm repair. Three patients died of multiple organ failure, after reoperation in two cases and infection in one case involving prior long-term corticosteroid therapy. Three patients developed paraplegia, including one who had undergone emergency treatment following rupture. Two patients required reoperation, for hematoma in one case and bowel necrosis in one. Four patients developed respiratory complications requiring artificial ventilation for more than 48 hr. During follow-up, two patients died from complications after repair of the proximal aorta and one patient required nephrectomy. Despite the extent of aneurysmal lesions and high frequency of association with visceral and supraaortic vessel lesions, the outcome of surgery in patients presenting with descending thoracic or thoracoabdominal aortic aneurysm in association with Takayasu disease was satisfactory. PMID- 15534729 TI - Ten years experience of thoracic and thoracoabdominal aortic aneurysm surgical repair: lessons learned. AB - In the last few years, advances in surgical techniques and in organ protection adjuncts have improved outcomes in thoracic (TAA) and thoracoabdominal aortic aneurysm (TAAA) surgical repair, although mortality and morbidity are still noteworthy. The aim of the current retrospective study is to determine whether the use of adjuncts influenced mortality and morbidity rates. From 1993 to 2003 we performed 353 procedures for TAA (175 cases) and TAAA (178 cases). This series has been divided into two consecutive groups: in group I (from 1993 to 1997), distal aortic perfusion with left atriofemoral bypass and cerebrospinal fluid drainage were used selectively, and in group II (from 1998 to 2003), the adjuncts were used routinely (together with surgical techniques of less invasive approach in selected cases). Total in-hospital mortality rates were significantly different ( p < 0.05): 15.9% in group I and 8.6% in group II. The overall incidence of paraplegia or paraparesis in group I was 8.3% and in Group II it was 5.1%. Renal failure occurred in 9.6% of group I and in 4.1% of group II. The incidence of respiratory failure in group I was 28%, and was 17.9% in group II. Respiratory failure was significantly lower ( p < 0.05) in group II. The reduction in the incidence of renal failure and paraplegia in the two groups was nonsignificant. In conclusion, the use of adjuncts and our improved experience allowed us to achieve a significant improvement in mortality and major morbidity rates in the group of patients operated on after 1998. PMID- 15534730 TI - Aortoiliac stent graft infection: current problems and management. AB - Aortic stent graft infection is uncommon. Most cases have been described anecdotaly in single-case reports. After observing one case in our experience, we decided to review the literature and contact centers performing endovascular aortic repair to determine the frequency, risk factors, and current treatment of stent graft infection. The literature was reviewed and the authors of identified articles were contacted for further information. In addition, 40 centers specializing in endovascular treatment were contacted by means a dedicated questionnaire. A total of 65 aortic stent graft infections were identified, including 43 reported cases and 22 previously unpublished cases that were observed at specialized centers. Stent grafts were implanted in the aorta in 50 cases and in the iliac artery in 15 cases. The frequency of infection was 0.43%. The gender ratio was 4:1 (M:F). Twenty-three percent of patients had immunodeficiency factors. Placement was performed in an interventional radiology suite in 62.5% of cases and in a sterile operating theater in 37.5%. Also, 35.5% of patients underwent other vascular procedures during the course of study and 29.2% stent grafts benefited from adjuvant endovascular procedures. Infection was classified as low grade in 35.4% of patients and high grade in 64.6%. Thirty-one percent of infections were associated with aortoenteric fistula. The offending microorganism was Staphylococcus aureus in 54.5% of cases. Treatment was conservative in 18% of cases and surgical in 82%. Surgical treatment consisted of stent graft removal followed by either extraanatomical bypass (59.5%) or in situ prosthetic reconstruction (40.5%). Mortality was 18% overall, 36.4% after conservative treatment and 14% after surgical treatment ( p = 0.083). Mortality was 16% after surgical treatment with extraanatomical bypass vs. 5.8% surgical treatment with in situ reconstruction. From these results we conclude that stent graft infection is an uncommon occurrence associated with poorly defined risk factors. Surgical treatment with complete excision of the infected stent graft followed by in situ reconstruction provides the best outcome. Establishment of a multicenter register to record such complications is needed to confirm the findings of this study. PMID- 15534731 TI - Carotid endarterectomy: experience in 5425 cases. AB - From 1992 to December 2002, 3967 patients (2619 males; 1348 females) with a mean age of 68.4A years (range 32-92) underwent 5425 carotid endarterectomy (CE) procedures at our institute. Neurological history was positive for stroke in 1130 cases (21%) and for transient ischemic attack (TIA) in 2121 cases (39%). In 2174 cases (40%) patients were neurologically asymptomatic or presented nonspecific symptoms. Our current clinical protocol has been designed to optimize resources and reduce complications. Some of the major features, along with the respective percentages in this series, are as follows. Duplex scanning was performed at a validated laboratory as the principal preoperative exam (86.9%). Locoregional anesthesia and neurological monitoring were performed during carotid cross clamping (96.3%). Selective shunting was carried out with a Javid shunt (10.7%). The choice of surgical technique was made according to carotid anatomy and cerebral tolerance of cross-clamping. Those used were a standard technique (now abandoned, 12.1%), synthetic patching (46.4%), and eversion endarterectomy (41.5%). Intraoperative completion arteriography was routinely performed for eversion endarterectomy and only in dubious cases with other techniques. The option of staying in an postoperative intensive care unit (ICU) was available (selective use, 2%). In uncomplicated cases, early discharge (after 1.5 postoperative days) was considered safe. The overall perioperative mortality was 0.37% (20/5425). Causes of death were myocardial infarction in seven cases, ischemic stroke in six cases, hemorrhagic stroke in five cases, respiratory failure caused by cervical hematoma in one case, and wound infection in one case. Perioperative neurological morbidity was 1.31% (71/5425); there were 43 major and 28 minor strokes. In conclusion, CE is effective for stroke prevention when there is significant symptomatic and asymptomatic carotid stenosis, as low mortality and morbidity may be achieved in an experienced center. At our institute, the reduction of costs did not have negative consequences on the quality of the surgical care. PMID- 15534732 TI - The bypass flap: an innovative technique of distal revascularization--anatomical study and clinical application. AB - Combined surgery for placement of a distal venous bypass and a free flap enables successful treatment of tissue loss caused by ischemia. This complex surgery has limited indications. The multiple anastomoses on the same arterial axis increase the risk of thrombosis and a certain number of venous grafts are likely to undergo mid-term deterioration. Because of these difficulties, we propose a new concept: the bypass flap (BF), which is based on the harvesting of an arterial axis to provide an arterial graft and a free flap supplied by a collateral branch of the graft. The aim of the anatomic part of this study was to evaluate the length and diameter of the arterial graft and its tissue branch and to study the feasibility of the BF. Thirty-two anatomic preparations were made by intraarterial injection of Rhodosil in 16 cadavers. The arterial graft included the subscapular artery and the thoracodorsal artery. The flap consisted of the anterior serrate muscle supplied by the branch of that graft. The distribution, length, and diameter of the arteries were examined. The mean length of the arterial graft line maintaining diameter above 2 mm was 12.5 cm (8.5-15.5). Three clinical applications of the BF based on the thoracodorsal artery axis were performed on three patients with tissue loss caused by severe ischemia of the lower limb. No occlusion of the BF occurred and healing of the tissue loss was achieved after 4, 7, and 10 months, respectively. This technique has the advantage of decreasing vascular distal resistance, which may contribute to improvement of vessel reconstruction patency. It is simpler because the anastomoses are fewer and it presents the advantage of requiring only autologous arterial material of an appropriate diameter. PMID- 15534733 TI - Duplex arteriography prior to femoral-popliteal reconstruction in claudicants: a proposal for a new shortened protocol. AB - The standard preoperative duplex arteriography (DA) from the aorta to the pedal vessels is time consuming and may be unnecessary in patients presenting with calf claudication alone. The feasibility of a shortened protocol was evaluated. Of 286 femoral-popliteal reconstruction based on DA during the last 4A years, 79 (28%) were primary operations for calf claudication. Eliminating the aortoiliac portion of the test except for the distal external iliac artery and limiting the scanning of the infrapopliteal vessels to one or two arteries in the leg would significantly shorten the exam. To confirm the adequacy of the inflow tract, we relied on the common femoral artery Doppler waveform analysis and the intraoperative graft pressure upon completion of the bypass. Of the 79 primary femoral-popliteal bypasses, 53 (67%) had triphasic common femoral artery waveform and the remaining 26 had monophasic or biphasic waveforms. Three (6%) of the 53 femoral-popliteal bypasses in the former group had significant pressure gradients measured intraoperatively and were treated with iliac angioplasties and stents for unsuspected stenoses in 2 cases and a covered stent for a common iliac aneurysm in 1 case. Three, two, and one infrapopliteal vessel runoff was observed in 24 (45%), 16 (30%), and 9 (17%) extremities, respectively. Four patients (8%) had significant stenoses (>50%) or occlusion of all three infrapopliteal arteries. Eighty-one percent of the patients would have completed the short protocol had we scanned the peroneal artery initially. An additional 8% would have required scanning of a second vessel (anterior tibial) and only 11%, scanning of all three infrapopliteal vessels. The time interval for completion of short-protocol DA was significantly less than the time for the standard DA (16.2 A+/- 5.2A min vs. 35.1 A+/- 10.6 min) ( p < 0.01). We believe that the proposed short DA protocol combined with intraoperative graft pressure measurements can be used in 94% of the patients who have a patent popliteal artery (>/= 7 cm). It is a totally noninvasive approach that is particularly suitable for vascular technologists and surgeons who wish to start utilizing DA instead of contrast arteriography prior to infrainguinal reconstructions. However, the short protocol does not avert the need for completion arteriography of the inflow arteries and readiness to perform endovascular procedures to correct lesions not suspected by common femoral artery waveform analysis. PMID- 15534734 TI - Characterization and probability of upper extremity deep venous thrombosis. AB - The objective of this study was to characterize patient demographics, risk factors, and anatomic distribution of upper extremity deep venous thrombosis (UEDVT) to develop a probability model for diagnosis. A retrospective review of all patients who underwent color-flow duplex scanning (CDS) for clinically suspected acute UEDVT over a 5-year period was performed. Patient risk factors and clinical symptoms were evaluated as predictors. Technically adequate complete CDS of 177 upper extremities (UEs) of arms were reviewed. CDS scanning identified acute UE venous thrombosis in 53 (30%) of the arms examined with deep system involvement in 40 (23%). Of the UEs affected, the subclavian was involved in 64%, the axillary in 25%, the internal jugular in 32%, the brachial in 36%, the cephalic in 32%, and the basilic in 47%. Multivariate analysis identified limb tenderness (odds ratio 9.3), history of central venous catheterization (odds ratio 7.0), and malignancy (odds ratio 2.9) as positive predictors for UEDVT. Erythema (odds ratio 0.12) and suspected pulmonary embolism (odds ration 0.06) were identified as negative predictors. A predictive model was designed from these variables. The anatomic distribution of UEDVT obtained from this study is consistent with previous reviews. Potential positive and negative risk factors can be identified from which a predictive model can be designed. Use of this model can help focus clinical suspicion, improve color-flow duplex utilization, and provide timely treatment with anticoagulation. PMID- 15534735 TI - Thoracic outlet syndrome surgery: long-term functional results. AB - The treatment of thoracic outlet syndrome (TOS) is controversial and long-term results are poorly documented. This retrospective study was carried out to assess clinical outcome 2 years after TOS surgery and to determine predictive factors of outcome. Between 1979 and 1999, 155 patients underwent TOS surgery. Of these patients, 140 (90.3%) had a minimum follow-up of 2 years. Thirty-six (25.7%) patients underwent bilateral procedures. A total of 176 procedures were reviewed and served as a basis for study. Presenting symptoms were neurologic in 15 cases (8.5%), arm or hand ischemia occurred in 38 cases (21.6%), and venous compression or thrombosis in 27 cases (15.4%). In 96 cases (54.5%), symptoms were mixed. A transaxillary approach (107 cases) was chosen to address venous symptoms and minor arterial dysfunction. A supraclavicular approach (69 cases) was used when there were large abnormal bony structures, neurologic symptoms, and/or severe limb ischemia. The first rib was resected either extensively from its neck to the sternal attachment (54 cases) or partially, including the mid-rib and neck or the mid-rib and sternal attachment (121 cases), depending on symptoms, approach, and surgeon's choice. Whenever present, cervical ribs, anterior scalenus muscle, and various fibrous or muscular compressive structures were also removed. The 2-year functional outcome according to Derkash's classification was assessed by means of a phone survey. Pre-, per-, and immediate postoperative data were recorded and compared to functional outcomes. Mean follow-up was 7.5 A+/- 3.4 years (2a"19 years). Functional results were excellent, good, fair, and poor in 87 (49.4%), 61 (34.6%), 14 (8%),and 14 (8%) procedures respectively. Predictive factors of negative outcomes were acute ischemia ( p < 0.01), sensory or motor deficit ( p < 0.01), and poorly systematized neurological symptoms as presenting symptoms ( p < 0.05), and extended resection of the first rib ( p < 0.01) and severe postoperative complications ( p < 0.01). This series showed that the 2 year results of TOS surgery were satisfactory in the majority of cases. Patients suffering from poorly systematized neurological symptoms in the arm had poor results. This subset of patients should be denied surgery or at least informed that postoperative results might be disappointing. Partial first-rib resection and a careful technique avoiding postoperative complications were also factors in long-term success. PMID- 15534736 TI - Subclavian stenosis/occlusion in patients with subclavian steal and previous bypass of internal mammary interventricular anterior artery: medical or surgical treatment? AB - There are only a few published studies on the association between subclavian steal syndrome and ischemic heart disease. The objective of this report is to evaluate the efficacy of subclavian steno-occlusion treatment in patients with subclavian steal syndrome (SSS) and previous coronary bypass. Over the last 8A years we observed 207 patients who underwent left internal mammary artery intraventricular artery (LIMA-IVA) bypass graft. Of these, 31 patients were affected by steno-occlusion of the homolateral subclavian artery. Ten patients (group 1) showed latent vertebral-SSS and were pharmacologically treated. Seven patients (group 2) had an intermittent vertebral-SSS; four patients were treated with angioplasty and stent application and three were pharmacologically treated. Fourteen patients (group 3) with complete vertebral-SSS were treated with angioplasty and stent application or carotid-subclavian bypass graft. All patients were followed up every 3 months for a period of 5A years after the diagnosis. The first group of patients showed no angina and no sign of subclavian restenosis. In the second group only two patients, who were affected by angina, showed subclavian restenosis at angiography. In the third group only one patient underwent further angioplasty for restenosis. The results of this study show that the SSS may be an adverse event in patients with a LIMA-IVA bypass graft. Identification of the steal is essential to choose an appropriate therapeutic approach. PMID- 15534737 TI - Coagulopathy and hyperfibrinolysis in ruptured abdominal aortic aneurysm repair. AB - Perioperative hemorrhage is one of the principal causes of death in patients with ruptured abdominal aortic aneurysm (AAA). This study examines perioperative coagulation and fibrinolysis in patients undergoing ruptured AAA repair complicated by coagulopathy. Eight patients (8 men of median age 74, range 69-87, years) who developed clinical and laboratory evidence of coagulopathy during attempted repair of ruptured infrarenal AAA were prospectively studied. Platelet count, fibrinogen, clotting times, prothrombin fragment (PF) 1+2, and tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI) activities were measured preoperatively, immediately before, and 5 min and 24 hr after aortic declamping. Six patients died, three intraoperatively, one within 24 hr, and two in the late postoperative period. All patients had thrombocytopenia and prolonged clotting times intraoperatively with evidence of increased thrombin generation (as demonstrated by elevated PF 1+2). Five patients had increased systemic fibrinolysis (as demonstrated by elevated t-PA activity) preoperatively and/or before aortic declamping and all of these patients died. Three patients had perioperative inhibition of systemic fibrinolysis (as demonstrated by elevated PAI activity) and two survived. These data demonstrate that coagulopathy in ruptured AAA repair may be associated with a hyperfibrinolytic state. Further research is required to determine if (a) a causal relationship exists between hyperfibrinolysis and coagulopathy and (b) whether antifibrinolytic agents can improve outcome if targeted at this group of patients. PMID- 15534738 TI - Does post-stenotic dilatation enhance collateral flow? AB - Our goal was to investigate whether post-stenotic dilatation (PSD) enhances collateral blood flow. In vitro experiments and computer modeling analysis were used to study the flow through stenotic segments and through collateral channels in the presence and absence of PSD. Pulsatile blood flow was provided by a left heart simulator primed with glycerol or normal saline. Pressure gradients across the stenosis were measured at different "cardiac" outputs. Computer models were constructed to simulate the experiments. Flow patterns and pressure drop across the stenosis were determined for a steady flow of 3 L/min. We observed that PSD was associated with a larger pressure drop across the stenosis than the absence of PSD when the flow was occurring through the stenosis only. There was, however, no difference in the pressure drop between the two geometries when the flow was occurring through both the stenotic orifice and the collateral channels when saline solution was used, but a small pressure difference prevailed for glycerol. At all different geometries there was considerable turbulence at PSD, and PSD geometry was found to be either at a disadvantage or at no advantage when compared to the tapered geometry for the total flow past the stenosis. The PSD geometry, however, enhanced the flow through the collateral while the flow through the orifice decreased concomitantly, resulting in no net increase in the total flow. This was true for any proportion of the total flow going through the collateral channels. For the total flow past the stenosis, PSD does not offer a benefit over tapered geometry. PMID- 15534739 TI - Vena cubiti media for creation of a loop-shaped arteriovenous fistula at the elbow region. AB - Hemodialysis access is one of the major routes, for patients who need long-term hemodialysis. In this report, we tried a new secondary arteriovenous fistula technique. Between January 2000 and June 2003, hemodialysis access was created in more than 650 patients with end-stage renal failure at our institution. In 15 cases a loop-shaped hemodialysis access was created between the brachial artery and vena cubiti media as a second choice. The youngest patient was 12 years old, whereas the oldest was 75. The mean follow-up period (A+/-SD) for these patients was 10.4 A+/- 2.53A months. Twelve patients are still using these fistulas successfully for hemodialysis (86.6%), whereas one patient died 7A months after the operation (6.7%). In one patient, the arteriovenous fistula failed 6A months after the operation (6.7%). Thrombosis due to hypotension occurred in two cases (13.3%) and this was resolved by simple thrombectomy. One patient had an aneurysm over the vena cubiti media (6.6%). None of the patients had an infection. These results suggest that an end-to-side anastomosis between the brachial artery and, vena cubiti media can be used as a second choice for hemodialysis access in patients with end-stage renal disease. PMID- 15534740 TI - Use of autologous superficial femoral artery in surgery for aortic prosthesis infection. AB - The surgical management of aortic prosthesis infection remains an enormous challenge for the vascular surgeon and a critical issue for the patient's well being and life. Several techniques for its management have been devised and employed over the years, but none of them has been totally satisfactory. Thus new alternatives are needed. We present the clinical case of an infected aortic prosthesis in a 41-year-old man that was complicated by duodenal and ureteral erosion. The autologous superficial femoral artery was used successfully in the treatment of this most demanding situation. PMID- 15534741 TI - Common origins of carotid and subclavian arterial systems: report of a rare aortic arch variant. AB - An aberrant right subclavian artery (aSA) arising from the proximal descending aorta is one of the most common anomalies of the aortic arch. We present our experience with an asymptomatic atypical aSA variant found during routine anatomic dissection. This aortic arch variant had two branches, the first being a bicarotid trunk and the second being a common trunk for both subclavian arteries. The right subclavian artery traveled behind the esophagus to reach the right upper extremity, thus forming an incomplete vascular ring around the trachea and the esophagus. The literature has been silent about the existence of this exact aSA variation. A plausible embryologic explanation is provided. An aSA is rarely symptomatic, but when symptoms do occur and intervention is warranted, it is important for surgeons and radiologists alike to be aware of the vascular anomalies that may potentially coexist with this entity. The surgical and endovascular options associated with this unique vascular anomaly are also discussed. PMID- 15534742 TI - Extraanatomic ascending-to-descending aortic bypass graft for recurrent, diffuse aortic coarctation. AB - Surgical correction of recurrent coarctation of the aorta presents a formidable technical challenge. If the lesion is diffuse or has long-segment aortic hypoplasia, it can be more difficult to manage. We present a successful case of repair using an extraanatomic bypass graft from the innominate artery to descending aorta. This technique can be an effective and easy approach for selected cases of complex or reoperative aortic obstruction. PMID- 15534743 TI - Connection between a long-standing traumatic arteriovenous fistula and development of aneurysmal disease. AB - Long-standing peripheral arteriovenous fistulas (AVFs) are always accompanied by ectasia of the proximal arteries. In the literature, traumatic fistulas of the lower limbs are frequently reported to be associated with iliac and even infrarenal aortic aneurysms; however, no study dealing with associated visceral aneurysms has been published. We report a case in which a traumatic AVF was accompanied by the late development of not only an infrarenal aortic aneurysm but also both superior mesenteric and right renal artery aneurysm. No causal relationship may be inferred between the tibial fistula and the other aneurysms, but this previously unreported finding does raise the question of a possible connection. PMID- 15534744 TI - Recurrent hemarthrosis of the knee caused by pseudoaneurysm of a composed femoropopliteal bypass graft. AB - We report on a case of recurrent hemarthrosis of the knee caused by a false aneurysm of a composed femoropopliteal bypass graft. Treatment consisted of resection and interposition grafting. The pathogenesis and treatment options of anastomotic false aneurysms are also reviewed briefly. PMID- 15534745 TI - Open versus endovascular repair of abdominal aortic aneurysms: what does each really cost? AB - Endovascular repair (EVAR) of abdominal aortic aneurysm (AAA) has emerged as an alternative to open repair (OR). The aim of this study was to compare the clinical outcomes and specific costs of these procedures since commercialization.A retrospective analysis of 119 consecutive infrarenal AAA repaired via an EVAR or an OR between July 2000 and September 2001 was performed. Patient charts were reviewed. Diagnostic-related group (DRG) classification and payer mix were identified. The hospital cost accounting system was accessed to obtain actual variable direct cost (AVDC) for the two groups. Percentages of the mean AVDC for the two groups were compared in the following cost categories: graft, operating room, radiology procedures and supplies, pharmacy, respiratory therapy, clinical laboratories, surgical floor, and monitored unit. Hospital profit margins were determined. Fifty-five patients underwent EVAR and 64 patients underwent OR. Mean aneurysm size was 5.5A cm (EVAR) and 6.1A cm (OR). Mean intensive care unit (ICU) stay was 0.09 days for EVAR vs. 3.5 days for OR ( p < 0.05). Mean length of stay (LOS) was 1.96 days for EVAR vs. 7.3 days for OR ( p < 0.05). Reimbursement was based on DRG 110, 47.3% in the EVAR and 79.7% in the OR group ( p < 0.05), and DRG 111, 50.9% in the EVAR group and 12.5% in the OR group ( p < 0.05). The payer mix showed no significant differences between the two groups. Mean AVDC for EVAR was 1.74 times that of OR. Significant differences in the distribution of costs were found in the following: graft costs (58% vs. 6.3%, p < 0.05), radiology procedures and supplies (3.9% vs. 0.1%, p < 0.05), pharmacy (1.9% vs. 10.5%, p < 0.05), and monitored unit (7.3% vs. 24.65%, p < 0.05) comparing EVAR vs. OR, respectively. Median cost of an endovascular graft was 22.4 times that of the standard graft for OR. Average hospital profit margins for an EVAR case was 49.5% vs. 88.6% for OR. Despite significant differences in monitored unit utilization, pharmacy services, and respiratory therapy services by the OR group, the cost of EVAR is appreciably more expensive. Furthermore, increased DRG reimbursement, and decreased ICU use and LOS do not compensate for the cost of EVAR. The main cost of EVAR is the cost of the graft itself. Hospital profit margins are acceptable with both the EVAR and OR procedures at this time; however, with proposed reductions in reimbursement, the ability to cover the cost of this new technology may be threatened. PMID- 15534746 TI - Robotics in vascular surgery. PMID- 15534747 TI - An enabling procedure: laparoscopic cholecystectomy. PMID- 15534748 TI - Best of the hotline. PMID- 15534749 TI - The tail shouldn't wag the dog. PMID- 15534750 TI - SCAR R&D Symposium 2003: comparing the efficacy of 5-MP CRT versus 3-MP LCD in the evaluation of interstitial lung disease. AB - The efficacy of two medical-grade, self-calibrating, gray scale displays were compared with regard to impact on sensitivity and specificity for the detection of interstitial lung disease (ILD) on computed radiographs (CR). The displays were a 5-megapixel (MP) cathode ray tube (CRT) device and a 3-MP liquid crystal display (LCD). A sample consisting of 230 anteroposterior (AP), posteroanterior (PA), and lateral views of the chest with CT-proven findings characteristic for ILD as well as 80 normal images were compared. This double-blinded trial produced a sample sufficient to detect if the sensitivity of the LCD was 10% or more reduced (one-sided) from the "gold standard" CRT display. Both displays were calibrated to the DICOM gray scale standard and the coefficient of variation of the luminance function varied less than 2% during the study. Five board-certified radiologists specializing in thoracic radiology interpreted the sample on both displays and the intraobserver Az (area under the ROC curve) showed no significant correlation to the display used. In addition, an interobserver kappa analysis showed that the relative disagreement between any observer pair remained relatively constant between displays, and thus was display invariant. This study demonstrated there is no significant change in observer performance sensitivity on 5-MP CRT versus 3-MP LCD displays for CR examinations demonstrating ILD of the chest. PMID- 15534751 TI - A review of the automated detection of change in serial imaging studies of the brain. AB - Serial imaging is frequently performed on patients with diseases of the brain, to track and observe changes. Magnetic resonance imaging provides very detailed and rich information, and is therefore used frequently for this application. The data provided by MR can be so plentiful; however, that it obfuscates the information the radiologist seeks. A system which could reduce the large quantity of primitive data to a smaller and more informative subset of data, emphasizing change, would be useful. This article discusses motivating factors for the production of an automated process to this effect, and reviews the approaches of previous authors. The discussion is focused on brain tumors and multiple sclerosis, but many of the ideas are applicable to other disease processes, as well. PMID- 15534752 TI - Multidimensional analysis: a management tool for monitoring HIPAA compliance and departmental performance. AB - Most RIS and PACS systems include extensive auditing capabilities as part of their security model, but inspecting those audit logs to obtain useful information can be a daunting task. Manual analysis of audit trails, though cumbersome, is often resorted to because of the difficulty to construct queries to extract complex information from the audit logs. The approach proposed by the authors uses standard off-the-shelf multidimensional analysis software tools to assist the PACS/RIS administrator and/or security officer in analyzing those audit logs to identify and scrutinize suspicious events. Large amounts of data can be quickly reviewed and graphical analysis tools help explore system utilization. While additional efforts are required to fully satisfy the demands of the ever-increasing security and confidentiality pressures, multidimensional analysis tools are a practical step toward actually using the information that is already being captured in the systems' audit logs. In addition, once the work is performed to capture and manipulate the audit logs into a viable format for the multidimensional analysis tool, it is relatively easy to extend the system to incorporate other pertinent data, thereby enabling the ongoing analysis of other aspects of the department's workflow. PMID- 15534753 TI - OsiriX: an open-source software for navigating in multidimensional DICOM images. AB - A multidimensional image navigation and display software was designed for display and interpretation of large sets of multidimensional and multimodality images such as combined PET-CT studies. The software is developed in Objective-C on a Macintosh platform under the MacOS X operating system using the GNUstep development environment. It also benefits from the extremely fast and optimized 3D graphic capabilities of the OpenGL graphic standard widely used for computer games optimized for taking advantage of any hardware graphic accelerator boards available. In the design of the software special attention was given to adapt the user interface to the specific and complex tasks of navigating through large sets of image data. An interactive jog-wheel device widely used in the video and movie industry was implemented to allow users to navigate in the different dimensions of an image set much faster than with a traditional mouse or on-screen cursors and sliders. The program can easily be adapted for very specific tasks that require a limited number of functions, by adding and removing tools from the program's toolbar and avoiding an overwhelming number of unnecessary tools and functions. The processing and image rendering tools of the software are based on the open-source libraries ITK and VTK. This ensures that all new developments in image processing that could emerge from other academic institutions using these libraries can be directly ported to the OsiriX program. OsiriX is provided free of charge under the GNU open-source licensing agreement at http://homepage.mac.com/rossetantoine/osirix. PMID- 15534754 TI - A mobile phone integrated health care delivery system of medical images. AB - With the growing computing capability of mobile phones, a handy mobile controller is developed for accessing the picture archiving and communication system (PACS) to enhance image management for clinicians with nearly no restriction in time and location using various wireless communication modes. The PACS is an integrated system for the distribution and archival of medical images that are acquired by different imaging modalities such as CT (computed tomography) scanners, CR (computed radiography) units, DR (digital radiography) units, US (ultrasonography) scanners, and MR (magnetic resonance) scanners. The mobile controller allows image management of the PACS including display, worklisting, query and retrieval of medical images in DICOM format. In this mobile system, a server program is developed in a PACS Web server which serves as an interface for client programs in the mobile phone and the enterprise PACS for image distribution in hospitals. The application processing is performed on the server side to reduce computational loading in the mobile device. The communication method of mobile phones can be adapted to multiple wireless environments in Hong Kong. This allows greater feasibility to accommodate the rapidly changing communication technology. No complicated computer hardware or software is necessary. Using a mobile phone embedded with the mobile controller client program, this system would serve as a tool for heath care and medical professionals to improve the efficiency of the health care services by speedy delivery of image information. This is particularly important in case of urgent consultation, and it allows health care workers better use of the time for patient care. PMID- 15534755 TI - Integrating RIS/PACS: the web-based solution at University Hospital of Ribeirao Preto, Brazil. AB - Integration of a Radiology Information System (RIS) and a Picture Archiving and Communication System (PACS) reduces the risk of inconsistent data by reducing interfaces among databases that contain largely redundant information. Furthermore, RIS/PACS integration is the basis for a filmless radiology facility. Web technology is based on international standards and supplies the main features for the RIS/PACS integration task based on a client/server model. This article presents a web-based system developed to manage and distribute images and diagnostic information within the University Hospital of Ribeirao Preto (HCRP) at the University of Sao Paulo. PMID- 15534757 TI - Pilot trial of salbutamol in central core and multi-minicore diseases. AB - Several studies have documented positive effects of beta-adrenergic agonists on human skeletal muscle with regard to muscle mass and strength. The aim of this pilot study was to evaluate the effect of the beta2-agonist salbutamol (albuterol) in a group of children with central core disease and multi-minicore disease. Thirteen patients, 8 with central core disease (mean age 17.5 years) and 5 with minicore disease (mean age 13.6 years) received oral salbutamol at a dose of 2 mg four times a day. Measures of efficacy were the change from baseline at 3 and 6 months in muscle strength, assessed by MRC score, myometry, functional measures and forced vital capacity. Statistical analysis was performed using repeated measures ANOVA (significance level < 0.05). Two patients with central core disease stopped the medication after one month because they did not notice any improvement and another one with minicore disease after 4 months because of increased tremors and palpitations. The remaining ten (6 with central core and 4 with minicore disease) completed the course of salbutamol without any significant adverse effects. There were significant increases in myometry, MRC scores and forced vital capacity between baseline and the six-month assessments. For both myometry and MRC the difference was already significant at 3 months and this was associated with a significant increase in functional abilities assessed with a structured functional scale. Our results suggest that salbutamol was overall well tolerated and might be beneficial in both central core and minicore patients. Larger prospective randomised, double-blind, placebo-controlled trials with salbutamol will be needed to confirm these preliminary findings. PMID- 15534758 TI - Acute peripheral facial palsy in Lyme disease -- a distal neuritis at the infection site. AB - AIM: Children with acute peripheral facial palsy have often suffered tick bites and/or erythema migrans in the head/neck region on the same side. With respect to the pathogenesis of neuroborreliosis this topographical association was investigated in an animal model. METHODS: A Borrelia garinii strain, isolated from the CSF of a child with acute facial palsy, was injected in 9 rats intracutaneously in the right subauricular region. Infected rats were examined for clinical symptoms of Lyme disease, the spread of the spirochetes was investigated by PCR of necropsies (facial nerves, trigeminus nerves, heart, brain, skin) up to 47 days after infection. The nerve tissues were investigated by histology, immunohistochemistry and electron microscopy. RESULTS: None of the rats developed a facial palsy or other symptoms of Lyme disease. Borrelia DNA was found in the heart after 5 days and in the brain after 7 days of infection up to the end of investigation (47 days), as well as in the ipsilateral peripheral nerves after 7 to 33 days. Borrelia was detected by electron microscopy near endoneural vessels of the facial nerve. Peri-, epi-, and endoneural infiltrations of macrophages, plasma cells and B cells characterized an inflammation of the facial and trigeminus nerves ipsilateral to the infection site. CONCLUSION: An infection with Borrelia garinii in the subauricular region induces an ipsilateral neuritis of peripheral nerves. The particular vulnerability of the human facial nerve may be a result of its long intraosseus course. Thus, an inflammatory edema may injure the nerve in the canalis facialis. PMID- 15534759 TI - The evaluation of autonomic nervous function in a patient with hereditary sensory and autonomic neuropathy type IV with novel mutations of the TRKA gene. AB - We report on a 10-year-old girl with anhidrosis and insensibility to pain, but no severe mental retardation or self-mutilation, diagnosed as hereditary sensory and autonomic neuropathy type IV (HSAN IV). Genetic analysis of her TRKA gene, which is responsible for HSAN IV, revealed two novel missense mutations in the tyrosine kinase domain. Cardiovascular autonomic nervous system function tests showed normal muscle sympathetic nerve activity associated with arterial baroreflex, reduced skin sympathetic nerve activity in the second and fifth fingers and palms, and abnormal circadian rhythm of cardiovascular autonomic nervous system. These findings have never before been reported in HSAN IV and may provide a clue to the neurological pathophysiology of this disease. PMID- 15534760 TI - Diffusion-weighted imaging findings in juvenile metachromatic leukodystrophy. AB - Magnetic resonance (MR) imaging has an important role in the diagnosis of metachromatic leukodystrophy (MLD). We report diffusion-weighted MR imaging (DWI) findings of four cases of juvenile type MLD. DWI showed restricted diffusion lines with greater areas of increased diffusion in three patients and widespread increased diffusion in one patient. This variability in DWI findings can be related to the histological stage of the disease at the time of imaging, ranging from intracellular metachromatic material accumulation to breakdown of myelin membranes. PMID- 15534761 TI - Hyperechogenicity of the thalamus and basal ganglia in very preterm infants: radiological findings and short-term neurological outcome. AB - Cerebral ultrasound of preterm infants may show diffuse, bilateral, hyperechogenic "haze" over the thalami and basal ganglia (hyperechogenicity BGT). We explored whether this could be a pathological phenomenon. All cerebral ultrasound examinations performed in 2001 on infants < 35 weeks of age were reviewed. This resulted in a hyperechogenicity and non-hyperechogenicity group. The character of the hyperechogenicity BGT and the presence of concomitant brain lesions were noted. Detailed clinical and follow-up data from a selected group of infants < 32 weeks were reviewed and compared between the 2 groups. The incidence of hyperechogenicity BGT was 11 % (39/359) in infants < 35 weeks and 26 % (37/143) in infants < 32 weeks. Birth weight and gestational age were significantly lower and clinical course was more complicated in the hyperechogenicity group. Concomitant brain lesions were always present. In 12/39 infants with hyperechogenicity BGT, MRI (always performed for other reasons) was available, showing signal intensity changes in thalamic region in 5 infants. The neurological outcome at term was less favorable in the hyperechogenicity group, but similar at 1 year. Thus hyperechogenicity BGT mainly occurred in very small, sick infants and was always associated with cerebral pathology. MRI did not consistently show abnormalities in the thalamic region. It was not associated with a poorer outcome at 1 year. PMID- 15534762 TI - Hypovitaminosis D-related myopathy in immigrant teenagers. AB - INTRODUCTION: Vitamin D deficiency resulting in a limb-girdle muscle weakness was diagnosed in three veiled immigrant teenage girls. PATIENTS: Three girls had a progressive muscle weakness and pain during a period varying from 6 months to two years. On examination limb girdle muscle weakness, predominantly of the lower extremities, without other neurological abnormalities was found. Serum examination showed a decreased level of vitamin D and phosphate and an increased alkaline phosphatase, and in two girls decreased calcium and increased parathyroid hormone levels were found. After supplementation with vitamin D, the pain subsided and muscle strength increased within weeks. Serum examination of the female relatives revealed eight persons with hypovitaminosis D, without any complaints. CONCLUSIONS: Vitamin D deficiency can result in a limb-girdle myopathy in veiled immigrant teenagers in the Netherlands. Vitamin D supplementation leads to rapid recovery of the muscle strength. The female relatives of these patients should be examined too. PMID- 15534763 TI - Alternating hemiplegia of childhood: no mutations in the second familial hemiplegic migraine gene ATP1A2. AB - Alternating hemiplegia of childhood (AHC) is a rare disorder mainly characterised by attacks of hemiplegia and mental retardation. AHC has often been associated with migraine. Previously, we have excluded the involvement of the familial hemiplegic migraine (FHM) CACNA1A gene in four patients with AHC. A second gene for FHM was discovered recently: the ATP1A2 gene on chromosome 1q23, coding for the alpha 2 subunit of Na+,K+-ATPase. We performed a mutation analysis of the ATP1A2 gene in six patients, using direct sequencing, but found no mutations in any of the 23 exons. Other cerebral ion channel genes remain candidate genes for AHC. PMID- 15534764 TI - Late infantile onset krabbe disease in siblings with cortical degeneration and absence of cerebral globoid cells. AB - Krabbe disease, a disorder caused by the deficiency of lysosomal galactosylceramidase, is typically associated with cerebral white matter degeneration, cortical sparing, accumulation of macrophages ("globoid cells"), and ultrastructural needle-shaped inclusions. Two sisters presented with progressive neurological deterioration beginning before the age of 2.5 years. The first, who died at the age of 9 years, exhibited profound destruction of cerebral white matter with sparing of subcortical fibers but no globoid cells. The brain of the second, who died at the age of 15 years and who had a proven galactosylceramidase deficiency, exhibited white matter destruction, previously undescribed circumscribed spongiform cortical degeneration (postcentral, inferior temporal, cingulate), and cerebellar atrophy, but no globoid cells. The peripheral nerve biopsies from both girls exhibited typical needle-shaped inclusions in Schwann cells. These observations confirm the rare reports that Krabbe disease is not always associated with globoid cells in the brain. Psychosine, which accumulates in the brain, might be toxic to cortical neurons following prolonged survival. The reason for the regional susceptibility in the cerebral cortex is unknown. PMID- 15534765 TI - Nemaline rods and complex I deficiency in three infants with hypotonia, motor delay and failure to thrive. AB - Three infants are described who had nemaline rods on muscle biopsy and isolated deficiency of complex I of the respiratory chain on biochemical analysis. They all manifested failure to thrive from birth, and hypotonia and muscle weakness within the first three months of life. Different genetic defects leading to isolated complex I deficiency have been described associated with a variety of morphological changes on muscle biopsy, but rods have not been described. Nemaline rods have been secondary phenomena in a number of conditions, as well as being the primary abnormality in nemaline myopathy. However, the combination of nemaline rods and complex I deficiency is an association not previously reported. PMID- 15534766 TI - Large-cell medulloblastoma in Aicardi syndrome. Case report and literature review. AB - An eight-year-old girl with Aicardi syndrome (AIC) developed signs of increased intracranial pressure. A clinical and radiological investigation revealed a tumor in the posterior fossa, which was resected. The histopathological diagnosis was large-cell medulloblastoma. Eight months later, she died of a local recurrence, despite treatment with chemotherapy and radiotherapy according to a PNET protocol. In addition to the growth of a large-cell medulloblastoma at the location of the primary tumor and the meningeal spread of the tumor, the autopsy revealed major cortical and subcortical malformations of the brain. Various benign (e.g., plexus papillomas) and malignant tumors (angiosarcoma, embryonic carcinoma, and hepatoblastoma) have been reported in connection with Aicardi syndrome. A genetic analysis of AIC suggests that the mutation is localized on the distal part of the short arm of the X chromosome, an area that may be of importance for tumor development. This is the first report of a primary malignant brain tumor -- large-cell medulloblastoma -- in a patient with Aicardi syndrome. PMID- 15534767 TI - Short-chain acyl-CoA dehydrogenase deficiency associated with early onset severe axonal neuropathy. AB - Two unusual cases of axonal neuropathy associated with short-chain acyl-CoA dehydrogenase (SCAD) deficiency are described. These two unrelated infants presented with profound generalised weakness, particularly affecting the upper limbs. Clinical examination revealed generalised peripheral hypotonia and weakness, with absent deep tendon reflexes. An axonal polyneuropathy was confirmed on electromyogram (EMG) and nerve conduction studies (NCS) and, following an extensive metabolic screen, an acylcarnitine and organic acid profile consistent with a short-chain fatty acid beta-oxidation defect was found. In both cases, SCAD deficiency was confirmed by enzyme analysis. Genetic analysis showed the presence of common gene variations in the SCAD gene. SCAD deficiency is a rare disorder with a wide clinical phenotype. SCAD deficiency associated with axonal neuropathy has not previously been reported. As highlighted in these cases, it may be necessary to include axonal neuropathy as a presenting feature of SCAD. PMID- 15534768 TI - [Avian influenza--transmitted to people more often than expected]. PMID- 15534769 TI - [Inhaled corticoids--standardized procedures make possible better comparisons]. PMID- 15534770 TI - [Cystic fibrosis--additional non-invasive ventilation improves breath function]. PMID- 15534771 TI - [Pulmonary embolism: subcutaneous treatment with low-molecular-weight heparin?]. PMID- 15534772 TI - [Endobronchial lipoma: successful therapy by bronchoscopical laser resection vs. surgery]. AB - BACKGROUND: Endobronchial lipoma is a very rare benign lung tumor. Therapeutic alternatives are the interventional bronchoscopic laser resection or surgical treatment. METHODS: From 1996 to 2002 we diagnosed 16 patients with endobronchial lipoma in our institution. We examined the clinical data of the patients and their influence on the individual patient treatment. RESULTS: 14 (88 %) out of the 16 patients (60 +/- 14 y.) were male. Clinical symptoms were cough (81 %), dyspnea (75 %), pulmonary infiltration (63 %) and fever (31 %). In the right lung 11 lipomas (69 %) were located, in the left lung 5 lipomas (31 %). The distribution to main bronchus, upper lobe, middle lobe and lower lobe bronchus were 2 (12.5 %), 6 (37.5 %), 2 (12.5 %) und 6 (37.5 %). 14 (88 %) lipomas were found in the central airways, 2 were located in the periphery of the lung. All lipomas in the central airways were successfully treated by bronchoscopic laser therapy with a mean of 1.1 +/- 0.3 laser sessions. One patient died of concomitant bronchial carcinoma before therapy. Surgical treatment was only necessary in a very peripherally located huge lipoma and a lipoma with extensive parenchym destruction. CONCLUSIONS: Bronchoscopic laser resection should be considered as first line therapy in cases of endobronchial lipoma. It is safe, successful, rapid and less expensive compared to surgical resection. Surgery should only be indicated in patients with lipomas with very peripherally localisation or parenchym destruction, extrabronchial growth, suspected malignancy or technical problems at bronchoscopic resection. PMID- 15534773 TI - [Tuberculosis induced by Bacillus Calmette-Guerin immuno-prophylaxis -- case study]. AB - We describe a case of miliary tuberculosis induced by Bacillus Calmette-Guerin (BCG) as a complication of an infection after BCG-instillation therapy into the bladder because of bladder carcinoma. Bacilli surely entered blood circulation via an urethral lesion because of a difficult catheterisation. The 60 year old patient was administered to the hospital with septic temperature 4 four days after BCG instillation. CT showed a miliary patten and the diagnosis was confirmed by bronchoscopy: transbronchial biopsy showed granulomatous infiltration and an acid-fast rod-bacterium was detected in bronchial slime. Symptoms vanished after a consequent antituberculous triple therapy regime and the patient left hospital in a good general state of health. PMID- 15534774 TI - [Current diagnostic approach to pleural effusion]. AB - Pleural effusion is a common pneumologic and interdisciplinary problem. Transudate/exsudate discrimination of the pleural fluid by thoracentesis remains the diagnostic basic algorithm. Regardless of a number of new markers, classical LIGHT's criteria comprising the pleural fluid protein- and LDH-values (or their serum ratio respectively) reveal the highest potency with an overall accuracy of 95 %. Expansion to cholesterol-determination (triplet test) may be helpful to identify transudates in indeterminate cases. The need for further local diagnostic evaluation is then usually restricted to exudates. Bacterial pleurisy, malignant and tuberculous effusion are the principal differential diagnoses. With the use of a variety of conventional biochemical, cytologic, immunologic and microbiologic investigations, thoracentesis will allow- or substantially narrow diagnosis of exudates in about 70 %, with novel cell biological markers in some conditions up to 90 %. In bacterial pleurisy thoracentesis provides information directly relevant to management in terms of local interventions. It also constitutes a platform for more invasive imaging- or endoscopy-guided investigations with a focus on medical thoracoscopy (pleuroscopy). Blind needle biopsy is diagnostic in a range of 40 - 70 % both in malignancy and inflammatory disease, thoracoscopy may clarify exudative conditions in about 95 %. Thus malignancy may be specifically diagnosed in 97 % of cases, tuberculous effusion in virtually 100 %. The value of thoracoscopy is augmented by interventional options including complete evacuation of the pleural cavity, eventually followed by talc pleurodesis ("poudrage") in recurrent effusions or adhesiolysis, irrigation and fibrinolysis protocols in certain inflammatory conditions. These combined features as accomplished in local anesthesia on a remarkably high safety level characterise medical thoracoscopy as a gold standard tool for the management of pleural disease even in comparison to more elaborate surgical procedures. PMID- 15534775 TI - [80 years "the magic mountain by Thomas Mann"]. PMID- 15534776 TI - [In Memoriam Prof. Dr. med. Friedrich Trendelenburg]. PMID- 15534777 TI - Combined reconstruction of the diabetic foot including revascularization and free tissue transfer. AB - Diabetic patients, presenting with both peripheral vascular disease and large soft-tissue defects, are too often treated by primary amputation. A combined revascularization and free-tissue transfer procedure can extend limb salvage in these patients. The authors report their experience over 4 years with 19 diabetic patients with peripheral vascular disease and large soft-tissue defects of the foot requiring free-tissue transfer. Although there was a 100 percent flap survival, early local wound problems occurred in three patients (16.6 percent). The recurrence rate was about 18.7 percent, but no complementary flap procedures were mandatory. With a mean follow-up of 38 months (range: 23 to 55 months), the limb salvage rate was 94.4 percent. Although there was one limb loss and one patient with ambulation difficulties, 16 patients (84.2 percent) were fully rehabilitated and were able to function independently. Despite a rather small series, this study confirms that in selected diabetic patients, a combined approach of vascular and reconstructive surgeons can reduce the limb amputation rate with acceptable complication rates. This combined approach offers major benefits to these patients, especially stable coverage and preservation of ambulation, and should always be considered before amputation. PMID- 15534778 TI - Deep circumflex iliac artery (DCIA) free flap without DCIA: report of a unique case. AB - The iliac crest free flap is a reliable source of cancellous bone, muscle, and skin. The vascularization of this flap arises from the deep circumflex iliac artery (DCIA) which allegedly is always present. The authors report a unique case of successful microvascular transplantation of an iliac crest osteomyocutaneous free flap in a patient in whom the DCIA and DCIV were absent. PMID- 15534779 TI - Advantages of using intercostal vessels as the recipients for free flaps covering lumbar defects. AB - Anatomic conditions in the lumbar region can complicate procedures for covering defects. In particular, a free flap is often required when the defect is large, in which case suitable recipient vessels must be found to insure revascularization. This report concerns a 50-year-old woman with multiple basal cell carcinomas in the lumbar radiodermatitis zone, who underwent a large resection from D10 to S2. The defect was repaired using a free latissimus dorsi flap revascularized by microvascular anastomosis to the 8(th) intercostal pedicle. The advantages of using these recipient vessels are then considered relative to reports in the literature. PMID- 15534780 TI - Successful correction of severe contracture of the palm using arterialized venous flaps. AB - Severe contracture of the palm causes great difficulty in the performance of the tasks required in daily life. Resurfacing with skin grafts may be sufficient for treating mild and small contractures; but severe and extensive contractures involving the joints, tendons, and neurovascular bundles should be covered with thin flaps. The authors present four cases of successful reconstruction of severe and extensive contractures of the palm using large arterialized venous flaps. PMID- 15534781 TI - A novel model for the study of peripheral-nerve regeneration following common nerve injury paradigms. AB - Recent advances in molecular neurobiology include the development of transgenic mice that express genes encoding fluorescent proteins under neuron-specific promoters (XFP mice). These mice have been used in the field of developmental neurobiology, but use has expanded to include the study of peripheral-nerve axonal regeneration subsequent to crush or unrepaired transection injuries. This report presents a transgenic mouse, which differs from previously reported and commercially available mice, in that enhanced yellow fluorescent protein expression (EYFP) is driven by the human thy1 promoter (hThy1). Motor and sensory peripheral nerves in these mice appear a bright yellow-green under fluorescent microscopy. This study tracks nerve regeneration in live animals using a serial imaging system. It also introduces a novel model for examining the clinically relevant nerve-injury paradigms of tibial nerve transection repaired with primary neurorrhaphy or graft, and end-to-side neurorrhaphy. Live-animal serial nerve imaging is compared with wet-mount fluorescent microscopy and histomorphometry in the same nerve specimens. The use of transgenic mice that strongly express EYFP in their peripheral neurons, coupled with serial nerve imaging, provide an important methodology for studying the heterogeneous nature of axonal elongation following peripheral-nerve injuries. PMID- 15534782 TI - Prevention of denervation atrophy by nerve implantation. AB - Prevention of denervation atrophy in skeletal muscle by nerve implantation to a muscle belly was studied in a rat model. In the animals' legs, the tibial nerve branch innervating the lateral head of the gastrocnemius muscle was transected, and subsequently repaired by either simple nerve suture(n = 20) or interpositional nerve grafting (n = 20). The tibial nerve branch innervating the medial head was dissected out. The peroneal nerve was transected and implanted directly to the lateral head of the gastrocnemius muscle on one side and, as a control, the nerve on the other side was cut. After 6 months, isometric tension of the gastrocnemius muscle was greater on the implantation side than on the control side in the nerve-suture group ( p < 0.05), but no tension difference between the two sides was observed in the nerve-graft group. The effects of neurotization by an implanted nerve may compete with those by a repaired nerve, when neurotization of the latter is delayed. Only when the time interval between neurotization by an implanted nerve and reinnervation by a repaired original nerve is short, does nerve implantation attenuate denervation atrophy. PMID- 15534783 TI - Flap prefabrication and prelamination with tissue-engineered cartilage. AB - In reconstructive surgery, the integration of tissue-engineered cartilage in a prefabricated free flap may make it possible to generate flaps combining a variety of tissue components, to meet the special requirements of particular defects. One aim of the present study was to investigate prefabrication of a microvascular free flap by implanting a vessel loop under a skin flap in a rabbit model. A second aim was to report on the authors' preliminary experiences in prelaminating prefabricated flaps with autologous tissue-engineered cartilage, in terms of matrix development, inflammatory reaction, and host-tissue interaction. The flap was prefabricated by implanting a vessel loop under a random-pattern abdominal skin flap. The tissue-engineered cartilage constructs were made by isolating chondrocytes from auricular biopsies. Following a period of amplification, the cells were seeded onto a non-woven scaffold made of a hyaluronic-acid derivative and cultivated for 2 weeks. One cell-biomaterial construct was placed beneath the prefabicated flap, and two additional constructs were placed subcutaneously and intramuscularly. In addition, a biomaterial sample without cells was placed subcutaneously to provide a control. All implanted specimens were left in position for 6 or 12 weeks. Neovascularization in the prefabricated flap and biomaterial construct was analyzed by angiography. After explantation, the specimens were examined by histologic and immunohistochemical methods. The prefabricated flaps showed a well-developed network of blood vessels between the implanted vessel loop and the original random-pattern blood supply. The tissue-engineered constructs remained stable in size and showed signs of tissue similar to hyaline cartilage, as evidenced by the expression of cartilage specific collagen type II and proteoglycans. No inflammatory reactions were observed. The physiologic environment of the autologous rabbit model provided favorable conditions for matrix deposition and maturation of the cell-biomaterial constructs. These initial results demonstrated the potential of prefabricating an axial perfused flap, combined with tissue-engineered cartilage, thus creating functionally competent tissue components for reconstructive surgery with minimal donor-site morbidity. PMID- 15534784 TI - Ultrastructural analysis of peripheral-nerve regeneration within a nerve conduit. AB - The purpose of this study was to observe the cellular components of regenerating peripheral nerves within a nerve conduit. Rat sciatic nerves were placed in a silicone conduit with a 5-mm gap between nerve endings. At weekly intervals for 6 weeks, 70-nm sections of nerve tissue from the conduit were obtained for ultrastructural observation. The principal cellular components by the end of the first week were macrophages and fibroblasts. By the end of the second week, both myelinated and unmyelinated nerve fibers began to pass through the entire conduit. By the end of the fifth week, nerve fibers were present at various levels of maturity, with no evidence of inflammatory or immunologic response. By the end of the sixth week, the percentage of nerve fibers was 86 percent of the cellular components. This analysis provides cellular data on which to base additional research regarding functional outcomes when using nerve conduits. PMID- 15534786 TI - Molecular basis of congenital and acquired long QT syndromes. PMID- 15534787 TI - Preclinical strategies to assess QT liability and torsadogenic potential of new drugs: the role of experimental models. AB - The recognition of QT prolongation and torsade de pointes (TdP) in humans has resulted in the re-labeling of some drugs and the removal of others from the market in the past decade. Recent regulatory guidelines have recommended a battery of preclinical tests to assess a new drug for the QT liability in humans. The assessment includes the effect of a drug on: 1) the ionic current in stable cell lines expressing hERG channel; 2) action potential duration (APD) measured in isolated ventricular tissues; 3) the QTc interval and TdP in animals in vivo; and 4) APD, the QT interval, transmural dispersion of repolarization (TDR) and TdP potential in the isolated arterially-perfused ventricular wedge preparation. Because a noncardiac drug with an incidence of TdP even less than 0.1% can be potentially removed from the market, the experimental models used for preclinical testing have to be high sensitive and specific to the signals related to TdP. Among available experimental models, the rabbit left ventricle wedge preparation exhibits a high sensitivity and a high specificity in the identification of compounds positive and negative for QT prolongation and TdP. This is attributed to the fact that the preparation demonstrates strong signals related to QT prolongation in response to even a weaker QT prolonging agent. Signals specifically pertinent to the development of TdP, ie, early afterdepolarization (EAD) and an increase in TDR can be detected as well. The preclinical data obtained from the wedge preparation correlate well with clinical outcomes. PMID- 15534788 TI - Arrhythmogenic mechanisms of QT prolonging drugs: is QT prolongation really the problem? AB - Torsade de Pointes (TdP) is an atypical ventricular arrhythmias associated with the acquired and congenital forms of the long QT syndrome. The substrate for the arrhythmia develops as a consequence of the amplification of electrical heterogeneities intrinsic to the ventricular myocardium. These heterogeneities exist because of differences in the time course of repolarization of the three predominant cell types that make up the ventricular myocardium, giving rise to transmural voltage gradients and a dispersion of repolarization responsible for the inscription of the electrocardiographic T wave. A wide variety of drugs are capable of reducing net repolarizing current and thus amplifying the intrinsic spatial dispersion of repolarization, so as to create the substrate for the development of re-entry. The result is a prolongation of the QT interval, abnormal T waves, and development of polymorphic re-entrant ventricular tachycardia displaying characteristics of TdP. Recent studies demonstrate that prolongation of the QT interval is not the sole determinant of the potential of a drug to cause TdP; agents that do not increase transmural dispersion of repolarization have little or no potential to induce the arrhythmia despite their ability to prolong the QT interval. Moreover, drugs such as sodium pentobarbital, which reduce transmural dispersion of repolarization, can diminish the likelihood of TdP, despite their ability to prolong the QT interval. PMID- 15534789 TI - Errors and misconceptions in ECG measurement used for the detection of drug induced QT interval prolongation. AB - Drug-induced changes in cardiac repolarisation receive substantial attention by regulatory agencies. Since there are presently no established accurate possibilities of testing the propensity to torsade induction during clinical drug development, the regulators require drug-related QT interval changes to be thoroughly investigated with almost all new pharmacological agents. Small QT interval changes are easy to miss and the regulators therefore expect the relevant studies to be very precise. Such a precision is not easy to achieve and different strategies have been proposed. The purpose of this article is to review the most frequent misconceptions and errors in the electrocardiogram handling and measurements related to the detection to drug-induced QT interval changes. Specifically, the article discusses (a) the possibilities of automatic measurement by standard electrocardiographic equipment, (b) the danger of casual measurement by central laboratories handling the electrocardiograms, (c) the selection of recording leads for QT interval measurement, and (d) the problem of the so-called heart rate hysteresis of the QT interval adaptation. Suggestions are made for drug developers of what study design and quality control aspects are needed to avoid the most frequent imprecisions. PMID- 15534790 TI - A mechanistic approach to assess the proarrhythmic risk of QT-prolonging drugs in preclinical pharmacologic studies. AB - Drugs with diverse structures and from several therapeutic classes are reported to increase the risk that a patient will experience ventricular tachyarrhythmias (e.g., torsades de pointes [TdP]) during drug therapy. This review discusses the use of preclinical assays to assess the risk that a QT-prolonging drug will cause TdP. The mechanisms underlying the development of TdP and the factors that increase the risk of TdP are described and applied to the design of preclinical experimental models for detection of proarrhythmic drug actions. Recommended assays, conditions, and preparations for preclinical assessment of the drug induced risk to TdP are given. No single preparation can simulate all conditions that cause TdP in patients. However, the assays described herein are capable of detecting the proarrhythmic effects of currently used drugs, even when these effects are reported to be extremely rare in clinical practice. PMID- 15534791 TI - Regulatory advice on evaluation of the proarrhythmic potential of drugs. PMID- 15534798 TI - Monolayer cell cultures as model systems for studying paroxysmal atrial fibrillation. PMID- 15534799 TI - Potential role of body surface ECG mapping for localization of atrial fibrillation trigger sites. AB - Catheter ablation has revolutionized the clinical management of atrial fibrillation (AF) by offering a curative treatment option for this highly prevalent arrhythmia. Ablation therapy is aimed at electrical isolation of the pulmonary veins (PVs) as a means to prevent rapidly firing focal activation within the PVs from penetrating into the left atrium (LA) and initiate reentrant wavelet propagation. However, non-PV AF trigger sites may be present and lead to unsuccessful ablation or post-ablation AF recurrences. Infrequent trigger firing and the difficulty or inability to induce focal trigger activity in the electrophysiology laboratory limits invasive catheter-based mapping of non-PV trigger sites. Identification of AF trigger sites using the surface electrocardiogram (ECG) P wave morphology is feasible but conventional 12-lead scalar recordings do not offer the resolving power to provide discrete regional localization to potentially target catheter ablation. The present paper includes a review of preliminary clinical data on the use of a 65-lead ECG mapping system (Resolution Medical, Inc) for the non-invasive localization of AF trigger sites. This method utilizes a unique previously developed reference database of 34 mean paced P wave integral map patterns which are each specific to activation arising from a discrete segment in the LA and right atrium (RA). Trigger site localization is obtained by matching the P wave integral map morphology of a premature atrial contraction (PAC) with the reference database of 34 mean paced P wave integral map patterns. PMID- 15534800 TI - Wireless remote monitoring for patients with atrial tachyarrhythmias. PMID- 15534801 TI - Ablation of atrial fibrillation: energy sources and navigation tools: a review. PMID- 15534802 TI - Annotated ECG waveform data at FDA. PMID- 15534803 TI - Advances in ambulatory monitoring: regulatory considerations. AB - Conventional ambulatory electrocardiogram (ECG) (Holter) monitoring involves 2 or 3 surface leads recorded with electrode positions and signal characteristics that are different from diagnostic quality 12-lead ECGs due to the limitations imposed by technology on the ambulatory recorders. The rapid pace of technological development for medical devices, particularly electrocardiography, has now enabled the recording of diagnostic quality 12-lead ECG waveforms for extended time periods. This capability allows Holter recording to become another source for diagnostic 12-lead ECG records on a par with other modalities such as resting ECG and exercise stress testing. Additionally, other diagnostic techniques such as S-T segment analysis and Q-T interval analysis that rely on diagnostic quality waveforms can now be applied. All of these enhancements to the traditional Holter modality have altered the regulatory perspective of these devices, since the enhancements may represent a new intended use for the device. PMID- 15534804 TI - Permanently implantable diagnostic computerized devices: where is the regulatory rar? PMID- 15534805 TI - The triangular wave test for electrocardiographic devices: a historical perspective. AB - Baseline wander makes interpretation ECG recordings difficult, especially the assessment of ST deviation. Eliminating baseline wander without distorting the ST segment is a problem. The traditional high pass filter with a 0.5 Hz low frequency cutoff effectively suppresses baseline but introduces considerable distortion in the level of the ST segment. This distortion results from phase nonlinearities that occur when frequency content and wave amplitude change abruptly, as occurs where the end of the QRS complex meets the ST segment. Since the 1980s nonlinear, digital filters have been designed that can increase the low frequency cutoff without the introduction of phase distortion. The triangular wave test, first described in the 1990 AHA Recommendations, is an objective method for measuring the ability to suppress baseline wander without affecting the ST segment. This methodology was adopted in 3 American National Standards. PMID- 15534806 TI - Regulatory issues for computerized electrocardiographic devices. AB - Computerized electrocardiogram (ECG) devices are regulated in the U.S. by the FDA Center for Devices and Radiological Health (CDRH). This article aims to highlight the salient points of the FDA regulatory review process, including the important distinction between a "tool" claim and a "clinical" claim in the intended use of a computerized ECG device. Specifically, a tool claim relates to the ability of the device to accurately measure a certain ECG parameter, such as T-wave alternans (TWA), while a clinical claim imputes a particular health hazard associated with the identified parameter, such as increased risk of ventricular tachyarrhythmia or sudden death. Given that both types of claims are equally important and receive the same regulatory scrutiny, the manufacturer of a new ECG diagnostic device should consider the distinction and regulatory pathways for approval between the two types of claims discussed in this paper. PMID- 15534815 TI - A comparison of commonly used QT correction formulae: the effect of heart rate on the QTc of normal ECGs. AB - The corrected QT interval (QTc) is widely used in pharmaceutical studies and clinical practice. Bazett's QT correction formula is still the most popular, despite Simonson's warning in 1961 that it could not be recommended. Other QTc formulae, e.g. Fridericia, Framingham, and Hodges, are also used. This study compares these four formulae using 10,303 normal ECGs recorded from four US hospitals. QT intervals were measured by the same computer program on ECGs confirmed by physicians. The distributions of QTc based on Fridericia, Framingham, and Hodges formulae were similar but Bazett's was significantly wider. The global group QTc-heart rate (HR) correlation coefficients were calculated as Bazett 0.33, Fridericia 0.24, Framingham 0.26, and Hodges 0.11, with the uncorrected QT-HR correlation being 0.82. Overall by far, Hodges QTc is significantly less correlated with HR compared to the others. Certain subgroup correlations of gender and low, mid, or high HR show that one individual formula can out-perform the others, whereby automated selection of QT correction formula based on the patient's HR and gender could be implemented as another option in products. The upper normal limits of corrected QTc were determined by excluding the top 2% from the global distribution charts as follows: Bazett 483 ms, Fridericia 460 ms, Framingham 457 ms, and Hodges 457 ms. Whether for males and/or females, the middle range of HR from 60 to 99 bpm has similar upper normal limits of QTc for all formulae except Bazett. Numerous references recommend 420 to 440 ms as the threshold for reporting prolonged QTc when using Bazett's formula. Based on this database, 30% of apparently normal ECGs would be reported as having abnormal QT intervals for the 440 ms threshold, or 10% if 460 ms is chosen, compared to <2% for the other formulae. It was also noted that QT has a linear trend with HR but not with RR. PMID- 15534816 TI - Surface ECG vector characteristics of organized and disorganized atrial activity during atrial fibrillation. AB - BACKGROUND: The aim of this study was to examine atrial organization from vectorcardiograms (VCGs) derived from the surface ECG of atrial fibrillatory waves. METHODS: We retrieved ECGs recorded during ventricular asystole from 22 patients with AF undergoing ablation of the AV junction. The synthesized VCG of each f-wave cycle of each ECG and its plane of best fit, described by azimuth and elevation angles relative to the frontal plane, were computed. RESULTS: Fifteen of the 22 ECGs had at least 30% of the planes in a single 30-degree region of azimuth angles. Of these 15, 12 had the greatest percentage of planes with azimuth angles within 30 degrees of the sagittal plane; two were near the frontal plane; and one near the right anterior oblique plane. CONCLUSIONS: Varying degrees of organization were observed from VCGs of fibrillatory waves with the more organized examples having planes predominately near the sagittal plane. PMID- 15534817 TI - Modification of ACC/ESC criteria for acute myocardial infarction. AB - The American College of Cardiology (ACC) and European Society of Cardiology (ESC) recently proposed criteria for acute ST elevation myocardial infarction (STEMI). These criteria were based on STj >0.1 mV in limbs leads and V4-V6, or STj >0.2 mV in V1 to V3 with criteria being met in two contiguous leads. The criteria were neither age nor sex dependent and the aim of the present study was to evaluate whether or not improved STEMI criteria that were age and gender dependent could be developed. A training set of 789 ECGs from patients presenting with chest pain due to cardiac and other causes was available for study. Revised criteria for STEMI were developed using these data as well as ECGs from a normal adult population of 859 males and 637 females. A test set of ECGs was available in the form of 1220 ECGs recorded from a separate hospital from patients presenting with chest pain. 248 patients had an acute myocardial infarction on the basis of conventional clinical criteria while 972 did not. There was an improvement in sensitivity using the new criteria compared to the old criteria from 41.5% to 46.7% while specificity improved from 96.0% to 98.5%. Specificity in normals improved from 92.6% to 99.8%. The conclusion drawn is that while the ACC/ESC criteria are simple to apply, they are not particularly specific and can be improved by being supplemented by other ECG measures and optimized for age and sex. PMID- 15534818 TI - Microelectrode arrays: a new tool to measure embryonic heart activity. AB - The analysis of the sequential excitation of cardiac tissue is of high relevance, both for clinical pathophysiological purposes, eg, detection of sustained ventricular arrhythmias, as well as for experimental electrophysiology. Clinically, different technical approaches such as single electrode measurements and bipolar mapping electrode catheters have been used. In experimental setups several techniques to record cardiac activity have been proposed. Beside the well established intracellular current-clamp recordings of action potentials, recent studies have performed extracellularly activation sequence mapping or simultaneous multichannel action potential electrode array measurements. Measurement of extracellularly recorded field potentials (FPs) hereby especially provides detailed information about the origin and spread of excitation in the heart. A similar analytical approach for cardiac FPs advanced the analysis of excitation spread and arrhythmic activity in multicellular preparations like developmental differentiation tissue of mouse embryonic stem cells, multicellular preparations of isolated native embryonic cardiomyocytes or the embryonic heart in toto. The use of substrate-integrated Microelectrode Arrays (MEAs, Multi Channel Systems, Reutlingen, Germany) with 60 electrodes of 10-30 microm diameters on a 100-200 microm grid, coated with porous titanium nitride to minimize the impedance allows recording of FPs at a high signal to noise ratio. The possibility to electrically stimulate the tissue further expands the range of applications and bioassays. It may thus facilitate the evaluation of drug research providing detailed information about the interplay of the complex cardiac network, and might improve the predictability of physiological and pathophysiological conditions or drug effects in embryonic heart tissue. PMID- 15534820 TI - Recent advances in magnetocardiography. AB - New developments in instrumentation, in clinical application, as well as in data analysis and visualization have provided new momentum to magnetocardiography (MCG). On one hand robust, easy to use and budget-priced MCG-systems entered the market and are applied to a multi-centred clinical study. On the other hand highly sophisticated vectormagnetometer systems with >300 SQUID sensors are opening new perspectives in electrocardiology research. Several parameters have recently been introduced to evaluate MCG-signals in order to support diagnosis, therapy follow-up and risk stratification. Particularly interesting is the renaissance of the Hosaka-Cohen-transformation which allows to visualize so called pseudo current density (PCD) maps. A few examples are given to emphasise the value of these maps. PMID- 15534821 TI - MCG to ECG source differences: measurements and a two-dimensional computer model study. AB - In this work we combine body surface potential map (BSPM) and magnetocardiogram (MCG) measurements with computer simulations in order to elucidate a recent thesis that claims the orthogonality of the main sources of MCG and ECG. Body surface currents and MCG pseudo currents are calculated from measured BSPM and MCG data, respectively. In contrast to the MCG-ECG source orthogonality thesis, we observe the main orientation of the BSPM currents and MCG pseudo currents to have similar axis during most of the depolarization R wave. In an attempt to explain such measurements we simulate a 2D transmural slice of the left ventricle in contact to a volume conductor. The main magnetic source currents along the wave front are indeed orthogonal to the extracellular electric current. However, fiber orientation inhomogeneity through the ventricular wall, volume conductor interface, wave front shape and extra- and intracellular potential distributions, all distort the symmetry of the current loops that contain the wave front currents. The resulting asymmetry rotates the main axis of the pseudo MCG currents away from the orthogonal axis of the body surface currents. Thus, the simulation results could solve the apparent contradiction between the orthogonal source theory and the observed similar ECG and MCG main current axis. PMID- 15534819 TI - Determination of electrical properties of ES cell-derived cardiomyocytes using MEAs. AB - Pluripotent embryonic stem cells (ES cells) provide a novel tool to study cardiomyogenesis under in vitro conditions. This overview article focuses on the technical properties of extracellular recordings of ES cell-derived cardiomyocytes using Microelectrode Arrays (MEAs). It reviews recent experimental observations, in an effort to describe basic characteristics of field potentials (FPs) in the ES cell model of developing myocardium. ES cells kept in permanent culture are differentiated within aggregates ("embryoid bodies", EBs) in which among other cell types cardiomyocytes appear 3-4 days after plating. These form spontaneously beating clusters mostly consisting of expanded regions of cardiac cells connected with narrow tissue strands. To record the electrical activity of these contracting areas substrate-integrated MEAs consisting of 60 substrate integrated electrodes can be used. We previously investigated the influence of Na+-, K+- and Ca2+ channel blockers on the electrical signal generation and propagation as well as on the shape of FPs. We also used ES cell-derived cardiac myocytes as a multicellular in vitro model for cardiac development. Long-term recordings with a MEA enabled the examination of electrophysiological properties during the ongoing differentiation process. During time in culture the beating aggregate of cardiac myocytes differentiating from ES cells increased in size (7 fold). This change was accompanied by an increase of the beating frequency from 1 to 5 Hz and a decrease of the FP duration. Furthermore a shortening of the FP upstroke velocity could be observed concomitant with a functional segregation of slow upstroke velocities in the area of the pacemaker. Our data indicate a functional differentiation and segregation of the cells into pacemaker and myocard-like regions. This in vitro development of a three-dimensional heart like structure closely follows the development known from mouse embryonic heart. The preparation thus forms an ideal model to monitor the development of electrical activity in embryonic cardiac myocytes for wild type and genetically modified ES cells, thereby taking into account the functional differentiation of the tissue. Our data suggest that EBs plated on MEAs provide a suitable tool for pre screening of cardioactive substances. PMID- 15534822 TI - Optical mapping of propagation changes induced by elevated extracellular potassium ion concentration in genetically altered mouse hearts. AB - Diabetes is associated with high rates of cardiovascular disease and sudden death. Therefore, dissecting specific mechanisms, such as the effects of impaired insulin signaling on cardiac electrophysiology may lead to better diagnosis and treatment. Lack of insulin receptors in mouse myocytes has been shown to reduce repolarizing potassium currents and prolong action potential duration. We hypothesized that these changes would manifest as rate-related effects on electrical propagation in the intact heart. This study employed optical mapping to characterize propagation changes in intact mouse hearts with cardiomyocyte restricted knock out of insulin receptors (CIRKO). METHODS: Fluorescent signals emitted from excited Di-4-ANEPPS in isolated Langendorff perfused mouse hearts were recorded from the left ventricular epicardium using an 8 by 8 photo diode array. The study included hearts from 8 CIRKO mice and 8 wild type (WT) littermate controls. Hearts were stimulated from the right atrium or the left ventricle at basic cycle lengths ranging from 160 to 280 ms under normal conditions and then after 5 minutes of perfusion with elevated potassium ion concentration (9.4 mM). RESULTS: None of the 8 CIRKO hearts maintained regular responses to atrial stimulation at the 160 ms cycle length under normal conditions; however, all of the WT hearts were captured at this rate. Total activation time for a 4 mm by 4 mm area was longer for CIRKO hearts when compared with WT. Average epicardial conduction velocity was slower for the CIRKO when compared to WT. Propagation delay due to the presence of high [K+]e was significant in both CIRKO and WT mice, but significantly longer for the CIRKO hearts. CONCLUSIONS: These results show that in addition to reducing repolarization currents, impaired myocardial insulin signaling leads to impaired electrical impulse propagation particularly at increased heart rates. These data suggest a link between impaired myocardial insulin signaling and the increased risk of arrhythmia and sudden death in patients with diabetes. PMID- 15534824 TI - The ISCE ECG genome pilot challenge: a 2004 progress report. AB - The International Society for Computerized Electrocardiography (ISCE) "genome project" began in 2000 as an open-ended discussion of ECG database needs and opportunities. Cooperation within ISCE led to a "pilot challenge" of the database concept, which called for establishment of methodology for transmission, storage, and integrated re-analysis of digitized waveforms of three different ECG manufacturers. The present report documents the early implementation of that goal. PMID- 15534823 TI - Does cardiac pacing reproduce the mechanism of focal impulse initiation? AB - Stimulation of myocardium by either a native pacemaker or an artificial stimulus requires the initiation of a self-propagating wave of depolarization originating from the site of initial activation. In the present study we perform artificial stimulation at a site of focal discharge with the aim to compare the two mechanisms of impulse formation. High resolution epicardial mapping in senescent rat hearts provided examples of focal discharge during sinus rhythm at a single epicardial breakthrough (BKT) point emerging from an isolated Purkinje ventricular muscle junction (PMJ) site. Stimulation was also performed at the same BKT point and potential distributions recorded during spontaneous and artificial stimulation were compared. During excitation latency, the negative potential pattern was elongated perpendicularly to fiber direction at both pacing and BKT point, in agreement with virtual cathode membrane polarization predicted by the bidomain model during point stimulation. During impulse initiation, activation wave fronts were initially circular around pacing site or BKT point and then elongated along local fiber direction. The similarity between impulse initiation during focal discharge and point stimulation in cardiac muscle suggests that high resolution pace mapping studies can help to elucidate the mechanism of abnormal impulse formation. PMID- 15534833 TI - What's old, what's new--in nonarrhythmia clinical electrocardiography: plenary address. PMID- 15534834 TI - Sympathovagal imbalance in pediatric patients with neurocardiogenic syncope during asymptomatic time periods. AB - The pathophysiology of neurocardiogenic syncope (NCS) is only poorly understood. Several studies indicate that NCS is associated with an imbalance of the autonomic nervous system (ANS). We hypothesized that pediatric patients with NCS exhibit alterations of the neurovegetative status also during asymptomatic time periods. To test this hypothesis the non-invasive method of Heart Rate Variability (HRV)-Analysis was used. METHOD: Holter records (12 channel, 180 Hz; obtained during an asymptomatic 24-hour period) of 32 patients (18 male, 14 female, mean age 14.6 yrs) with a history of syncope and a positive Head-Up tilt (HUT) were compared to the recordings of 33 healthy control subjects (19 male, 14 female, mean age 13.5 yrs) with negative history of syncope and HUT. Time domain and frequency domain features were calculated. Analysis segments were divided in different signal duration groups (1, 3, 6, 24 hours). RESULTS: For all time scales the standard deviation of wavelet coefficients yields the best discrimination properties. Analysis of the total time duration (24h) shows only moderate discrimination (sensitivity 84%, specificity 45%) between patient and controls. However, analysis of 3 and 6 hour segments (starting and 6 a.m.) showed significant discrimination: sigma wave scale 13 (6-9 a.m.) sensitivity 78%, specificity 71%., sigma wave scale 11 (6-12 a.m.) sensitivity 84%, specificity 61%. The best combination of two parameters is denoted by SDNN and sigma wave scale 11 (6-12 a.m.) with sensitivity 70%, specificity 75%. CONCLUSION: The results indicate that during an asymptomatic 24-hour-period patients with NCS exhibit an imbalance of the ANS especially in the morning (6-12 a.m.). The findings suggest a time-dependent increased sympathetic or reduced vagal activity in pediatric patients with NCS. Patients could benefit from a time-adjusted medical therapy with beta-blocking agents. PMID- 15534835 TI - Simultaneous comparison of three derived 12-lead ECGs with standard ECG at rest and during percutaneous coronary occlusion. PMID- 15534837 TI - The feasibility of ST-segment monitoring with a subcutaneous device. AB - We have developed a subcutaneous cardiac monitor/alarm for patients at high risk that alerts companions or bystanders if cardiac arrest occurs. The leadless device has 4 electrodes mounted on its corners. An added feature of the implantable monitor/alarm is the ability to detect ST-segment deviations present in acute ischemia. We obtained acute ischemia electrocardiogram (ECG) data of 30 patients from the Utah 192-site body surface maps and determined the left precordial region where our device would be implanted. (The device is implanted at the location where its electrodes yield maximum QRS amplitude.) Unipolar body surface potentials from the chosen area were further interpolated so that closely spaced bipolar electrocardiograms can be derived by performing algebraic subtraction of these "unipolar" ECGs. Results show that the summation of the absolute values of ST-deviations from a pair of diagonal bipolar leads from the optimal subarea (3 x 6 cm rectangle) provides a measure comparable to the maximum ST-deviation among the 4 corner unipolar leads of the same area. This suggests that ST-segment deviations seen at neighboring precordial sites are sufficiently different that significant cancellation of these deviations does not occur in forming close bipolar ECGs. Our conclusion is that during acute ischemia, bipolar ECG recordings between neighboring precordial sites can present ST-segment deviations that are large enough for automatic detection in many patients. Furthermore, the same electrodes used by our subcutaneous device in monitoring R waves in arrhythmia detection can also be used in monitoring ST-segment deviations for acute ischemia detection. PMID- 15534838 TI - Tracking repolarization dynamics in real-life data. AB - Ambulatory (Holter) electrocardiographic recordings provide the tools for tracking temporal instabilities of repolarization during various daily activities. However, analysis of low-amplitude repolarization changes in this setting is challenging due to the presence of multiple artifacts, variable activity levels, and other uncontrolled factors. Here we compare performance of different methods for continuous analysis of repolarization dynamics using simulated signals and real-life Holter recordings. Selection of relatively stable segments with a low baseline drift and accurate correction of baseline wander constitute the first step in repolarization analysis. We describe application of adaptive filtering, which yields more accurate results than non-adaptive techniques. Because small (microvolt-level) residual baseline drifts can be a source of error in tracking repolarization changes, stability of isoelectrical segment has to be controlled. To compare robustness of spectral and time-domain techniques for tracking temporal repolarization instabilities (T-wave alternans, TWA), we used simulated signals with changing heart rate, variable levels of TWA, noise, phase shifts, spurious artifacts, and period-four oscillations. In addition, we compared performances of the inter-beat and intra-beat averaging techniques for tracking dynamics of T-wave alternans. Using the simulated signals and real-life Holter data, we showed that analysis of information both in time and frequency domains combined with control of baseline drifts (surrogate analysis) gives a more reliable estimate of the low-amplitude repolarization dynamics than each of these techniques alone. To summarize, dynamic tracking of low-amplitude repolarization changes in ambulatory recordings is possible during most of the recording time but requires accurate control of baseline wander and stability of isoelectrical segments. Analysis of time-frequency distributions embedded in repolarization dynamics facilitates detection of abrupt and transient repolarization instabilities, including changes in the level of T-wave alternans and slower periodicities. PMID- 15534839 TI - Magnetocardiography for pharmacology safety studies requiring high patient throughput and reliability. AB - Recent guideline drafts of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) underline the necessity to test nonantiarrhythmic drugs for their potential to prolong the QT or the corrected QT (QTc) interval. The implementation of these guidelines requires a large amount of ECG measurements on animals and humans in preclinical and clinical phases of the drug development process. We propose the use of magnetocardiography (MCG) as a complementary method with particular advantages in high-throughput studies, where signal quality and reliability are key factors. Our proposal is based on a review of recent MCG studies investigating the repolarization phase and results of methodological work assessing QT interval parameters from the MCG. The applicability of MCG for pre clinical in-vivo studies is demonstrated by the ease of measurement in unrestrained non-anesthetized rabbits, guinea pigs, and hamsters.. PMID- 15534840 TI - The dominant T wave. AB - This contribution discusses the dominant T wave, a concept providing an overall view on the repolarization currents of the ventricular myocardium. The dominant T wave is a weighted mean of the T waves on the body surface. It can be estimated from the matrix of sampled observed potentials. The timing of its peak reveals the mean of the repolarization times of the involved transmembrane potentials. Its amplitude is independent of the volume conductor properties of the tissues surrounding the heart. PMID- 15534841 TI - Spatial repolarization parameters for predicting cardiac death in the elderly. PMID- 15534842 TI - Post infarction risk stratification using the 3-D angle between QRS complex and T wave vectors. AB - Present experience with prospective identification of patients who might benefit from prophylactic antiarrhythmic intervention is restricted to risk stratification using left ventricular ejection fraction (LVEF). The precision of LVEF-based identification of high risk patients is neither highly sensitive nor highly specific. This study investigated risk stratification of 466 survivors of acute myocardial infarction (86 women, mean age 57.5 years) for whom a 5-year follow-up was available. During the follow-up 67 patients died and 24 of these events were sudden arrhythmic deaths. In addition to LVEF, patients were stratified by mean heart rate, heart rate variability and the slope of heart rate turbulence, all derived from 24-hour Holter recording obtained before hospital discharge, and by the 3D angle between QRS complex and T wave vectors (total cosine R-to-T, TCRT) obtained from digital resting electrocardiogram before hospital discharge. Individual risk characteristics and their combinations were evaluated by calculating the areas under the receiver operator characteristics (ROC). The bootstrap technology was used to investigate these statistically. For the stratification of both all cause mortality and sudden arrhythmic death, TCRT was the strongest risk stratifier (area under ROC of 0.6857 +/- 0.0367, and 0.7275 +/- 0.0544, respectively) that compared very favourably to LVEF (area under the ROC of 0.6610 +/- 0.0362 and 0.6346 +/- 0.0595, for all cause and arrhythmic death prediction, both P < 10(-10) for the comparison with TCRT). TCRT was also stronger in combination with other stratifiers, eg, TCRT + LVEF (area under ROC of 0.7631 +/- 0.0325 and 0.8057 +/- 0.0473, for all cause and arrhythmic death prediction) was stronger than mean heart rate + LVEF (area under ROC of 0.7396 +/- 0.0298 and 0.7673 +/- 0.0445, respectively, both P < 10(-10) for comparison with TCRT + LVEF). Hence the 3D QRS-T angle is a very powerful risk stratifier especially suited for the prediction of sudden arrhythmic death. It should be prospectively investigated in future trials of prophylactic antiarrhythmic interventions. PMID- 15534843 TI - Looking for prognostic information in the ST-T segment--is it really worth it? AB - The West of Scotland Coronary Prevention Study (WOSCOPS) provided baseline electrocardiograms (ECGs) on 6,595 men without a previous myocardial infarction who were followed for a mean of 4.9 years during which time all events, cardiovascular or otherwise, were recorded. Half of the study group was treated with a lipid lowering drug while the other half was randomly assigned to placebo. This study cohort afforded the opportunity to look at ECG morphology as a marker of risk. All 12-lead ECGs in the study were processed by the Glasgow Program and automated Minnesota Coding was also undertaken. All computer outputs were reviewed to exclude errors due to technically unsatisfactory recordings. Multiple variables were studied. Univariate and multivariate logistic regression analyses were undertaken to determine those electrocardiographic and clinical parameters of predictive value with respect to the primary endpoint of fatal or non fatal myocardial infarction. Those ST-T variables with additional prognostic value in the multivariate analysis, which included the clinical parameters, were used to develop a risk score. Although many ECG measures were of prognostic value in a univariate analysis, only rate, frontal T axis and T+ amplitude in lead I were of significance in a multivariate analysis which included clinical data. With respect to QT dispersion, while it was shown that there was an increased risk for those with QT dispersion exceeding 44 ms (RR 1.38, CI 1.02 - 1.81 P = 0.034) the receiver operating characteristic curve was virtually a straight line. The risk equation also demonstrated that there was as much prognostic value in the use of age and smoking history alone as there was in ECG plus age combined. The conclusion drawn is that the prognostic value of ECG variables has to be considered carefully in the light of other available data. PMID- 15534844 TI - Pre-hospital synthesized 12-lead ECG ischemia monitoring with trans-telephonic transmission in acute coronary syndromes: pilot study results of the ST SMART trial. AB - INTRODUCTION: The aim of the ST SMART trial is to determine whether prehospital ST monitoring with telephone transmission to the target hospital will improve hospital time to treatment in acute coronary syndromes. The present analysis reports results of the feasibility pilot study. METHODS: All patients calling 911 for chest pain in Santa Cruz County California were monitored with a synthesized 12-lead ECG. Prehospital ECGs were printed for clinical use in the experimental group; control group patient care used only ECGs recorded after hospital arrival. RESULTS: Five patients with non-ST elevation myocardial infarction or unstable angina had normal ECGs upon hospital arrival but evidence of ischemia in their prehospital ECGs. Three patients with ST elevation myocardial infarction were treated with primary percutaneous coronary intervention, with "door to balloon" times of 47 and 65 minutes in 2 experimental group patients and 148 minutes in the one control group patient. CONCLUSION: Prehospital ST monitoring appears feasible. Its potential to improve hospital time to diagnosis and treatment in acute coronary syndromes, and the clinical benefits of such improvement will be studied in the larger, ongoing ST SMART trial. PMID- 15534846 TI - Prehospital 80-LAD mapping: does it add significantly to the diagnosis of acute coronary syndromes? AB - Early detection of acute myocardial infarction (MI) is vital in the management of acute coronary syndromes (ACS). Hence we compared the diagnostic capability of the standard 12-lead electrocardiogram (ECG) with the 80-lead ECG body surface map (BSM) prehospital. METHODS: Consecutive patients (n = 294) presenting prehospital with ischemic type chest pain were included. All had an ECG and BSM pretreatment and a baseline and 12-hour cardiac troponin-T or I (cTnT or cTnI). Acute MI was defined as cTnT > 0.09 or cTnI > 0.1 ng/mL. Acute MI on the BSM was defined as ST elevation measured at the J-point, > or = 1 mm inferior/right ventricular/high right anterior/lateral regions, > or = 2 mm anterior region, > or = 0.5 mm posterior region. RESULTS: Acute MI occurred in 182/294 (62%) based on cTnT or I. ST elevation on the standard ECG predicted acute MI in 103 (sensitivity 57%, specificity 94%; c-statistic 0.73). The optimal model for the standard ECG included ST elevation, summed ST depression and past history of MI (c-statistic 0.82; Chi-square (Wald) 120.7, 3df). The BSM predicted acute MI in 146 (sensitivity 80%, specificity 92%; c-statistic 0.86). The optimal model for the BSM included BSM criteria for acute MI and past history of MI (c-statistic 0.91; Chi-square (Wald) 180.3, 2df). CONCLUSION: The 80-lead BSM is superior to the standard 12-lead ECG in predicting acute MI prehospital. PMID- 15534847 TI - Added value of new acute coronary syndrome computer algorithm for interpretation of prehospital electrocardiograms. AB - A new computerized acute coronary syndrome (ACS) computer algorithm has been developed with the aim of improving the electrocardiographic detection of acute myocardial ischemia and infarction in the emergency department (ED). The purpose of this study was to determine the added value of the new ACS algorithm in assisting ED physicians to obtain a more accurate diagnosis in patients with ACS. The new algorithm combines a rule-based decision tree, which uses well-known clinical criteria and a data-centered neural network model for more robust pattern recognition. Input parameters of the neural network model consist of morphology features of derived Frank X, Y, Z waveforms and the patient's gender and age. The neural network model was trained with electrocardiograms obtained from documented acute myocardial infarction patients at the Mayo Clinic who were a part of a research ACS database, which includes electrocardiograms (ECGs) of more than 5,000 individuals at hospital admission (1st ECG in the ED). The test set portion of the study was conducted in 2 steps: 1) One emergency physician and 1 cardiologist classified 1,902 clinically correlated out-of-hospital ECGs without seeing the interpretation statement from the algorithm into 1 of the following categories: 1) acute myocardial infarction, acute ischemia, or nonischemic; 2) After 9 months, the same 2 physicians classified the same group of ECGs but with the interpretation statement of the algorithm printed on the tracing. The results demonstrated that with the assistance of the new algorithm, the emergency physician and cardiologist improved their sensitivity of interpreting acute myocardial infarction by 50% and 26%, respectively, without a loss of specificity. The new algorithm also improved the emergency physician's acute ischemia interpretation sensitivity by 53% and still maintained a reasonable specificity (91%). The new ACS algorithm provides added value for improving acute ischemia and infarction detection in the ED. PMID- 15534848 TI - Optimal leads, estimation, and continuous monitoring improve detection of acute MI and transient ischemia. PMID- 15534851 TI - The molecular mechanism of fetal hemoglobin reactivation. AB - Increased levels of fetal hemoglobin (HbF) are clinically beneficial in patients with sickle cell disease. Hydroxurea fails to increase HbF in at least 25% of patients, and therefore, better drugs are needed. Recent clinical studies have shown that the DNA methyltransferase (DNMT) inhibitor decitabine effectively increased HbF in hydroxyurea-refractory patients. The rational use of DNMT inhibitors as therapeutic agents to reactivate HbF expression in patients with sickle cell disease is based on nearly 25 years of experimental evidence, reviewed in this article, that supports a fundamental role of DNA methylation in the silencing of gamma-globin gene expression in adults. PMID- 15534852 TI - Clinical studies with fetal hemoglobin-enhancing agents in sickle cell disease. AB - Fetal hemoglobin (HbF, alpha2gamma2) decreases polymerization of sickle hemoglobin, and high levels correlate with decreased morbidity and mortality in sickle cell disease (SCD). Therefore, a therapeutic goal for patients with SCD is pharmacologic reactivation of HbF. Decreased HbF production is associated with DNA methylation (by DNA methyltransferase [DNMT]) at the gamma-globin (HbF) gene promoter. The cytosine analogs 5-azacytidine and 5-aza-2'-deoxycytidine (decitabine) hypomethylate DNA by inhibiting DNMT. In early studies, 5 azacytidine produced significant HbF elevations in patients with thalassemia and SCD, but clinical development of this class of agent was halted after a poorly controlled animal study suggested that 5-azacytidine might be carcinogenic. However, the majority of preclinical studies with decitabine have suggested a chemopreventive rather than carcinogenic effect. Furthermore, decitabine, unlike 5-azacytidine, does not incorporate into RNA and is a more directed DNA hypomethylating agent. Therefore, we have pursued studies of decitabine to pharmacologically reactivate HbF in patients with SCD. In phase I/II studies, decitabine at DNA-hypomethylating, but noncytotoxic, doses was well tolerated and effective at increasing HbF and total hemoglobin levels both in patients who had and had not responded to prior hydroxyurea therapy. In treated patients, there were marked improvements in a range of surrogate clinical endpoints measuring red blood cell adhesion, endothelial damage, and coagulation pathway activity. Pharmacologic reactivation of HbF through DNA hypomethylation holds promise as an effective disease-modifying intervention for patients with SCD. Larger studies are required to confirm the safety and effectiveness of decitabine with chronic use, and to more clearly establish its role in patients with SCD. PMID- 15534853 TI - The role of fetal hemoglobin-enhancing agents in thalassemia. AB - Augmentation of fetal hemoglobin (HbF) synthesis can reduce the severity of beta thalassemia by improving the imbalance between alpha- and non-alpha-globin chains. However, previous clinical trials of pharmacologic induction of HbF in thalassemia produced inconsistent results. Striking responses in HbF and total hemoglobin synthesis were occasionally observed, but in most patients, the increase in gamma-globin synthesis was inadequate to influence the clinical course of thalassemia. A small number of patients treated with azacytidine demonstrated a consistent response, but the development of this drug was abandoned due to concerns over toxicity. Decitabine, an analog of azacytidine, is a more potent inhibitor of DNA methyltransferase, and may be less toxic in clinical use. Trials in sickle cell anemia have confirmed that decitabine is effective in patients refractory to hydroxyurea. The clinical trials of decitabine in thalassemia must carefully evaluate the dose and route of administration, and address concerns about long-term side effects. PMID- 15534854 TI - Issues regarding study design for initial clinical trials using decitabine. AB - Decitabine offers a reasonable treatment alternative to hydroxyurea for individuals with sickle cell disease who have experienced multiple painful episodes. Despite the promise that the use of decitabine holds, several issues linger with regards to early trial design, such as what age group should be included and what trial design should be used in studies assessing its safety and clinical efficacy. Other issues that must be considered include determination of which primary clinical outcomes should be evaluated, what route of administration should be tested, and to how to assess toxicity of this novel therapeutic agent. This article will discuss issues regarding the factors that must be considered prior to initiating a phase III trial of decitabine for individuals with sickle cell anemia. PMID- 15534856 TI - Management of chronic myeloid leukemia in France: a multicentered cross-sectional study on 538 patients. AB - PURPOSE: Little is known about the actual management and treatment of chronic myeloid leukemia (CML) in clinical practice, although there have been many recent changes, such as the introduction of imatinib. PATIENTS AND METHODS: A two-phase cross-sectional observational study with retrospective data collection was conducted in France. In the first phase information regarding health services treating patients with CML was collected. In the second phase, centers caring for 10 or more patients were asked to provide data regarding patients diagnosed with CML that had had a consultation or been hospitalized in the last 3 months. RESULTS: All French departments of hematology (n=218) were contacted by phone. The median number of patients followed per center is 6 (range 0--200). The median number of new patients seen during the last 12 months was 2 (range 0--60). In the second phase 538 patients were included, the sex ratio being 1.14 and median age 55. At the time of diagnosis, 96.8% (n=519) were in chronic phase, 2.2% (n=12) in accelerated phase and 0.9% (n=5) in blastic phase. Eighty-two percent (n=443) of the patients have been treated by interferon (IFN). Sixteen point 3% (n=87) of the patients received a bone marrow transplantation (BMT). Forty-six percent (n=236) of the patients were treated with imatinib. CONCLUSIONS: This is the first study providing detailed descriptive data concerning the use of medications and procedures in a large population of patients from the medical centers involved in treating CML patients in France. Further observational studies are needed to assess the impact of different treatment strategies and economic impact of CML care in France. PMID- 15534857 TI - A pilot study to build a database on seven anti-hypertensive drugs. AB - PURPOSE: In Japan, all patients are able to see freely any clinics or hospitals. So clinical data of all patients have been stored at clinics, hospitals and medical institutes respectively. These patients' clinical course data stocks have not been combined with one another. There is no large-scale database, which has been available and has played its role in complementing spontaneous adverse drug reaction (ADR) reporting system. We tried to build an original database using anti-hypertensive drugs' data from Drug Use Investigation conducted for the Japanese Drug Re-examination application by every pharmaceutical manufacturer in conformity with Japanese Pharmaceutical Affairs Law and Related Regulation. METHODS: The 43 565 case data of seven anti-hypertensive drugs (one Ca antagonist, one alpha-blocker, two beta-blockers, three ACE inhibitors) were kindly offered from seven manufacturers who were members of RAD-AR Council, Japan. After examining the data items and categories, they were standardized into common codes based on Japanese Drug Category Classification (JDCC), International Classification of Diseases 9 (ICD-9) and Japanese Adverse Drug Reaction Terminology (J-ART). As each manufacturer had a different coding method in accordance to manufacturer's own practice of data management, the original forms were divided into several datasets. The data processing and statistical analysis were conducted using Statistical Analysis System (SAS). RESULTS: (1) Technology and know-how to combine data coded by different methods were established for building a database that had never been tried in Japan. (2) The following are the by-products of the study: a) Onset of ADR concentrated in the early stage but onset of some disorders prevailed equally throughout the investigation period. b) Although the number of collected cases of anti-hypertensive drugs was 43 565, total number of administrated anti-hypertensive drugs reached to 70 714 because additional anti-hypertensive drugs were often used with subject drugs. CONCLUSION: There is no large-scale database of patients' clinical course in Japan. However, since the Japanese Drug Re-examination System started in 1979, almost eight million cases of Drug Use Investigation on about 700 drugs have been collected with enormous human power and huge expenditure for Japanese Drug Re examination application by pharmaceutical manufacturers. New and more appropriate information will be detected by the database, built using Drug Use Investigation data that were collected only for the Japanese Drug Re-examination application. PMID- 15534858 TI - Changes in cell shape and anchorage in relation to the restriction point. AB - The restriction point (R) separates the G1 phase of continuously cycling cells into two functionally different parts. The first part, G1-pm, represents the growth factor dependent post-mitotic interval from mitosis to R, which is of constant length (3-4 h). The second part, G1-ps, represents the growth factor independent, pre-S phase interval of G1 that lasts from R to S and that varies in time from 1 to 10 h. G1-pm cells rapidly exit (within 1 h) from the cell cycle and enter G0 as a response to serum withdrawal. The finding that R occurs at a set time after mitosis indicates that R may be related to the metabolic and/or structural changes that the cell underwent during the previous mitosis. We have recently shown that phosphorylation of the retinoblastoma tumor suppressor protein (pRb) is not the molecular mechanism behind R, as has been suggested previously. Here, we present an alternative explanation for R. In the present study, we applied a single cell approach using time-lapse analysis, which revealed that upon serum starvation the G1-pm cells rapidly underwent a transient change in cell shape from flat to spherical before exiting to G0. Platelet derived growth factor (PDGF) counteracted this change in shape and also prevented exit to G0 to the same extent. Furthermore epidermal growth factor (EGF) and insulin like growth factor (IGF-1), which only partially counteracted this change, only partially counteracts exit to G0. These data clearly indicate a direct link between change in cell shape and exit to G0 in G1-cells that have not passed R. PMID- 15534859 TI - Matricellular protein SPARC is translocated to the nuclei of immortalized murine lens epithelial cells. AB - The matricellular glycoprotein, secreted protein acidic and rich in cysteine (SPARC), has complex biological activities and is important for lens epithelial cell function and regulation of cataract formation. To understand how SPARC influences lens epithelial cell activity and homeostasis, we have studied the subcellular distribution of SPARC in murine lens epithelial cells in vitro. We demonstrate that endogenous SPARC is located in the cytoplasm of either quiescent or dividing lens epithelial cells in culture. However, cytoplasmic SPARC was translocated into the nuclei of immortalized lens epithelial cells upon a significant reduction of intracellular SPARC in these cells. Recombinant human (rh) SPARC added to the culture media was quickly and efficiently internalized into the cytosol of SPARC-null lens epithelial cells. Moreover, cytoplasmic rhSPARC was also translocated into the nucleus after exogenous rhSPARC was removed from the culture media. The translocation of SPARC into the nucleus was therefore triggered by the reduction of SPARC protein normally available to the cells. A mouse SPARC-EGFP chimeric fusion protein (70 kDa) was expressed in lens epithelial cells and 293-EBNA cells, and was observed both in the cytoplasm and culture medium, but not in the nucleus. SPARC does not appear to have a strong nuclear localization sequence. Alternatively, SPARC might pass through the nuclear pore complex by passive diffusion. SPARC therefore functions not only as an extracellular protein but also potentially as an intracellular protein to influence cellular activities and homeostasis. PMID- 15534861 TI - Different cation sensitivities and binding site domains of Na+-Ca2+-K+ and Na+ Ca2+ exchangers. AB - We examined inhibitory effects of external multivalent cations Ni(2+), Co(2+), Cd(2+), La(3+), Mg(2+), and Mn(2+) on reverse-mode exchange of the K(+)-dependent Na(+)/Ca(2+) exchanger NCKX2 and the K(+)-independent exchanger NCX1 expressed in CCL-39 cells by measuring the rate of Ca(2+) uptake with radioisotope tracer and electrophysiological techniques. The apparent affinities for block of Ca(2+) uptake by multivalent cations was higher in NCKX2 than NCX1, and the rank order of inhibitory potencies among these cations was different. Additional experiments also showed that external Li(+) stimulated reverse-mode exchange by NCX1, but not NCKX2 in the presence of 5 mM K(+). Thus, both exchangers exhibited differential sensitivities to not only K(+) but also many other external cations. We attempted to locate the putative binding sites within the alpha motifs for multivalent cations by site-directed mutagenesis experiments. The cation affinities of NCKX2 were altered by mutations of amino acid residues in the alpha-1 motif, but not by mutations in the alpha-2 motif. These results contrast with those for NCX1 where mutations in both alpha-1 and alpha-2 motifs have been shown previously to affect cation affinities. Susceptibility tests with sulfhydryl alkylating agents suggested that the alpha-1 and alpha-2 motifs are situated extracellularly and intracellularly, respectively, in both exchangers. A topological model is proposed in which the extracellular-facing alpha-1 motif forms an external cation binding site that includes key residues N203, G207C, and I209 in NCKX2, while both alpha-1 and alpha-2 motifs together form the binding sites in NCX1. PMID- 15534860 TI - Hepatocyte growth factor enhances protein phosphatase Cdc25A inhibitor compound 5 induced hepatoma cell growth inhibition via Akt-mediated MAPK pathway. AB - We have previously shown that Compound 5 (Cpd 5), an inhibitor of protein phosphatase Cdc25A, inhibits Hep3B human hepatoma cell growth. We now show that hepatocyte growth factor (HGF), a hepatocyte growth stimulant, can strongly enhance Cpd 5-induced growth inhibition in Hep3B cells, and this enhancement in cell growth inhibition is correlated with a much stronger ERK phosphorylation when compared to cells treated with Cpd 5 or HGF separately. We found that HGF/Cpd 5-induced ERK phosphorylation and cell growth inhibition were mediated by Akt (protein kinase B) pathway, since combination HGF/Cpd 5 treatment of Hep3B cells inhibited Akt phosphorylation at Ser-473 and its kinase activity, which led to the suppression of Raf-1 phosphorylation at Ser-259. The suppression of Raf-1 Ser-259 phosphorylation caused the induction of Raf-1 kinase activity, as well as hyper-ERK phosphorylation. Transient transfection of Hep3B cells with dominant negative Akt c-DNA further enhanced both Cpd 5- and HGF/Cpd 5-induced ERK phosphorylation, while over-expression of wild-type Akt c-DNA diminished their effects. In contrast, HGF antagonized the growth inhibitory actions of Cpd 5 on normal rat hepatocytes, thus showing a selective effect on tumor cells compared to normal cells. Our data suggest that Akt kinase negatively regulates MAPK activity at the Akt-Raf level. Suppression of Akt activity by either combination HGF/Cpd 5 treatment or by dominant negative Akt c-DNA transfection antagonizes the Akt inhibitory effect on Raf-1, resulting in an enhancement of Cpd 5-induced MAPK activation and cell growth inhibition. PMID- 15534862 TI - DNA adduct formation by 7H-dibenzo[c,g]carbazole and its tissue- and organ specific derivatives in Chinese hamster V79 cell lines stably expressing cytochrome P450 enzymes. AB - The cytochrome P4501A subfamily (CYP1A) is involved in the metabolic activation of 7H-dibenzo[c,g]carbazole (DBC) and its tissue- and organ-specific derivatives, N-methyldibenzo[c,g]carbazole (MeDBC)and 5,9-dimethyldibenzo[c,g]carbazole (diMeDBC). In this study, we have evaluated the relationship between the tissue specificity and (32)P-postlabeled adduct patterns produced by these compounds by using a panel of Chinese hamster V79 cell lines stably expressing human CYP1A1 and CYP1A2 and/or N-acetyltransferase. Treatment of the parental cell lines V79MZ and V79NH, which are devoid of any CYP activity, with DBC and its derivatives did not result in detectable adducts. The highest DNA adduct levels were found in CYP1A1-expressing V79MZh1A1 cells after DBC and MeDBC treatment (24.5 +/- 7.2 and 16.2 +/- 3.6 adducts/10(8) nucleotides, respectively). Exposure of this cell line to DBC resulted in five distinct spots, while six spots with different chromatographic mobilities were detected in MeDBC-treated cells. DiMeDBC produced only very low levels of DNA adducts in V79MZh1A1 cells. DBC and MeDBC formed relatively low levels of DNA adducts in CYP1A2-expressing V79MZh1A2 cells (0.7 +/ 0.2 and 2.1 +/- 1.2 adducts/10(8) nucleotides, respectively). DBC formed three weak spots and MeDBC five spots in V79MZh1A2 cells, and all the spots had different chromatographic mobilities. In contrast, diMeDBC did not induce any DNA adducts in these cells, although diMeDBC induced a significant dose-dependent increase in micronucleus frequency under similar treatment conditions (r = 0.76; P < 0.001). The significant increase in DNA damage in the Comet assay following incubation of exposed cells with a repair-specific endonuclease (Fpg protein) suggests that base modifications such as 8-oxodG or Fapy-adducts might be responsible for the genotoxicity of diMeDBC in V79MZh1A2 cells. The similarities between the DNA adduct patterns produced by DBC and MeDBC in V79MZh1A1 and V79MZh1A2 cells suggest that biotransformation mediated via CYP1A1 and CYP1A2 might depend on a PAH-type pathway involving the aromatic ring system. PMID- 15534863 TI - Thrombin peptide (TP508) treatment of rat growth plate cartilage cells promotes proliferation and retention of the chondrocytic phenotype while blocking terminal endochondral differentiation. AB - A synthetic peptide representing the receptor-binding domain of human thrombin (TP508, also known as Chrysalin) accelerates fracture repair in rats via endochondral ossification and promotes repair of rabbit cartilage defects. To understand how this peptide might stimulate cartilage and bone formation, we employed an established in vitro model of growth plate cartilage regulation. Rat costochondral cartilage resting zone and growth zone chondrocytes were treated with 0, 0.07, 0.7, or 7 microg/ml TP508 or a scrambled peptide, TP508-SP. Proliferation ([3H]-thymidine incorporation) was examined in pre-confluent cultures; effects on cell number, alkaline phosphatase activity, [35S]-sulfate incorporation, and responsiveness to vitamin D metabolites were tested using confluent cultures. TP508 did not affect proliferation of resting zone cells but it caused a dose-dependent increase in cell number and DNA synthesis of growth zone cells. Alkaline phosphatase specific activity of resting zone cells was reduced by TP508, whereas [35S]-sulfate incorporation was increased. Neither parameter was affected in growth zone cell cultures. TP508 treatment for 24 h did not induce resting zone cells to respond to 1alpha,25(OH)2D3, either with respect to alkaline phosphatase activity or proteoglycan production. In contrast, TP508 treatment reduced the stimulatory effect of 24R,25(OH)2D3 on alkaline phosphatase but it did not alter the stimulatory effect of 24R,25(OH)2D3 on [35S]-sulfate incorporation. In cultures treated for 48, 72, or 140 h with TP508, 1alpha,25(OH)2D3 restored alkaline phosphatase activity to control levels but did not stimulate activity over levels observed in untreated control cultures. The stimulatory effect of TP508 on [35S]-sulfate incorporation was evident up to 48 h post-confluence but at later time points, proteoglycan production was comparable to that seen in control cultures, control cultures challenged with 1alpha,25(OH)2D3, and cultures treated with TP508 followed by 1alpha,25(OH)2D3. TP508-SP had no effect on any of the parameters tested. These results indicate that TP508 exerts maturation specific effects on chondrocytes in the endochondral lineage, promoting cartilage extracellular matrix synthesis over endochondral differentiation in resting zone cells and proliferation over differentiation of growth zone cells. PMID- 15534864 TI - DNA damage in bronchial epithelial and mesothelial cells with and without associated crocidolite asbestos fibers. AB - Mesothelioma is induced almost exclusively by exposure to asbestos fibers. We have investigated whether the induction of DNA damage in human bronchial epithelial BEAS 2B cells and human mesothelial MeT 5A cells by crocidolite asbestos (2 microg/cm2) requires the presence of asbestos fibers in the cells. DNA damage was measured microscopically by the Comet assay, and the presence of fibers in the same cells was assessed using bright-field illumination. After treatment times of 6-72 hr, damage levels were, on the average, two times higher in cells with fibers than in cells without fibers. It was further found that DNA damage decreased with time in BEAS 2B cells both with and without fibers. No decrease in damage with time was seen in MeT 5A cells, suggesting that these mesothelial cells repair the initial damage poorly, lack induction of protective systems, or constantly produce high levels of damaging species. Our results indicate that crocidolite-treated human mesothelial MeT 5A and bronchial epithelial BEAS 2B cells show an elevated level of DNA damage if they contain a fiber. In comparison with epithelial BEAS 2B cells, mesothelial MeT 5A cells have more DNA damage after the crocidolite treatment and the damage is more persistent. PMID- 15534865 TI - PTCH polymorphism is associated with the rate of increase in basal cell carcinoma numbers during follow-up: preliminary data on the influence of an exon 12-exon 23 haplotype. AB - After first presentation with a basal cell carcinoma (BCC), patients demonstrate interindividual diversity in the rate of development of further BCCs (number/year of follow-up). The mechanism for this variation is unknown. In this study, we evaluated whether PTCH variants mediate this phenomenon. We used negative binomial regression analysis to identify associations between BCC numbers/year and host characteristics, parameters of exposure to ultraviolet radiation (UVR), and PTCH exon 12(1686) C/T, intron 15(2560+9) G/C, and exon 23(3944) C/T genotypes and haplotypes in 279 BCC cases who presented with an initial tumor on the head/neck. PTCH genotypes were not significantly associated with BCCs/year, although cases with two copies of the C1686-C3944 haplotype developed significantly fewer BCCs/year than those without this haplotype (rate ratio = 0.44; 95% CI = 0.27-0.71). Cases with one copy of T1686-T3944 developed more BCCs/year (rate ratio = 2.46; 95% CI = 1.27-3.97) than those without the haplotype. We found no significant associations between BCCs/year and the other PTCH haplotypes studied. We reexamined the association of C1686-C3944 with BCCs/year in a model that included UVR exposure parameters (sunburning in childhood, sunbathing score, intermittency of exposure between 40 and 60 years of age, exposure in hours/year) and skin type, gender, and age at first presentation. The association between C1686-C3944 and BCCs/year remained significant (rate ratio = 0.44; 95% CI = 0.26-0.73 for two copies of the haplotype). The data show that allelic variation in PTCH contributes to the rate of development of BCC. PMID- 15534866 TI - Corneal epithelial adhesion strength to tethered-protein/peptide modified hydrogel surfaces. AB - In this study, we investigated the suitability of microjet impingement for use on hydrogel materials to determine the cellular adhesion strength of corneal epithelial cells grown on novel hydrogels with extracellular matrix proteins (laminin and/or fibronectin) or a peptide sequence (fibronectin adhesion promoting peptide, FAP) tethered to their surface with poly(ethylene glycol) chains. The deformation of the hydrogel surface in response to the force of the microjet was analyzed both visually and mathematically. After the results of these experiments and calculations determined that no deformation occurred and that the pressure required for indentation (1.25 x 10(6) Pa) was three factors of 10 greater than the maximum pressure of the microjet, the relative mean adhesion strength of primary rabbit corneal epithelial cells grown on the novel poly(2 hydroxyethyl methacrylate-co-methacrylic acid) hydrogels was determined and compared with that of the same type of cells grown on control glass surfaces. Only confluent cell layers were tested. Cells grown on control glass surfaces adhered with a mean relative adhesion strength of 488 +/- 28 dynes/cm2. Under identical conditions, cells grown on laminin- and FAP-tethered hydrogel surfaces were unable to be removed, indicating an adhesion strength greater than 516 dynes/cm2. Cells grown on fibronectin- and fibronectin/laminin (1:1)-tethered surfaces showed significantly lower relative adhesion strengths (201 +/- 50 and 189 +/- 11 dynes/cm2, respectively), compared with laminin- and FAP-tethered surfaces (p = 0.001). Our results demonstrate that the microjet impingement method of cell adhesion analysis is applicable to hydrogel substrates. Additionally, analysis of our test surfaces indicates that fibronectin tethered to this hydrogel in the quantity and by the method used here does not induce stable ligand/receptor bonding to the epithelial cell membrane to the same degree as does laminin or FAP. PMID- 15534867 TI - Poly(dimethylsiloxane) thin films as biocompatible coatings for microfluidic devices: cell culture and flow studies with glial cells. AB - Oxygen plasma treatment of poly(dimethylsiloxane) (PDMS) thin films produced a hydrophilic surface that was biocompatible and resistant to biofouling in microfluidic studies. Thin film coatings of PDMS were previously developed to provide protection for semiconductor-based microoptical devices from rapid degradation by biofluids. However, the hydrophobic surface of native PDMS induced rapid clogging of microfluidic channels with glial cells. To evaluate the various issues of surface hydrophobicity and chemistry on material biocompatibility, we tested both native and oxidized PDMS (ox-PDMS) coatings as well as bare silicon and hydrophobic alkane and hydrophilic oligoethylene glycol silane monolayer coated under both cell culture and microfluidic studies. For the culture studies, the observed trend was that the hydrophilic surfaces supported cell adhesion and growth, whereas the hydrophobic ones were inhibitive. However, for the fluidic studies, a glass-silicon microfluidic device coated with the hydrophilic ox-PDMS had an unperturbed flow rate over 14 min of operation, whereas the uncoated device suffered a loss in rate of 12%, and the native PDMS coating showed a loss of nearly 40%. Possible protein modification of the surfaces from the culture medium also were examined with adsorbed films of albumin, collagen, and fibrinogen to evaluate their effect on cell adhesion. PMID- 15534868 TI - Chiral recognition by mass-resolved laser spectroscopy. AB - Chiral recognition is a fundamental phenomenon in life sciences, based on the enantioselective complexation of a chiral molecule with a chiral selector. The diastereomeric aggregates, formed by complexation, are held together by a different combination of intermolecular forces and are therefore endowed with different stability and reactivity. Determination of these forces, which are normally affected in the condensed phase by solvent and supramolecular interactions, requires the generation of the diastereomeric complexes in the isolated state and their spectroscopic investigation. This review deals with chiral recognition in the gas phase through the application of laser-resolved mass spectrometric techniques (R2PI-TOF and RET-MS). The measurement of the fragmentation thresholds of diastereomeric clusters by these techniques allows the determination of the nature of the intrinsic interactions, which control their formation and affect their stability and reactivity. PMID- 15534869 TI - Paradigmatic identification of MMP-2 and MT1-MMP activation systems in cardiac fibroblasts cultured as a monolayer. AB - Activations of MMP-2 and membrane type 1-matrix metalloproteinase (MT1-MMP) have been correlated with cell migration, a key cellular event in the wound healing and tissue remodeling. We have previously demonstrated furin-dependent MMP-2 and MT1-MMP activations induced by type I collagen in cardiac fibroblasts. To understand mechanistic aspects of the regulation of MMP-2 and MT1-MMP activations by potential non-matrix factor(s) in cardiac fibroblasts, in the present study, we examined the effects of various agents including concanavalin A (ConA), a proteolytic phenotype-producing agent. We showed that treatment of cells with ConA activated pro-MMP-2, and that this activation concurred with elevated levels of cellular MT1-MMP and TIMP-2. The presence of active MT1-MMP and 43 and 36 kDa processed forms of MT1-MMP in a fraction of intracellular proteins prepared from ConA-treated cells suggests the possible internalization of differential forms of MT1-MMP. The appearance of 36 kDa processed form of MT1-MMP in conditioned media prepared from ConA-treated cells indicates the possible extracellular release of the further processed MT1-MMP fragment. Inhibition of furin in ConA-treated cells attenuated pro-MT1-MMP processing and the cellular TIMP-2 level, plus it reduced cell-released active MMP-2 in a time-dependent manner. These results suggest the involvement of furin in the ConA-induced activations of MT1-MMP and MMP-2. Furthermore, the existence of furin inhibitor-insensitive pro- and active MMP-2 species associated with ConA-treated cells implies that a mechanism independent of furin may perhaps account for the binding of the MMP-2 species to the cells. Supplementary material for this article can be found at http://www.mrw.interscience.wiley.com/suppmat/0730 2312/suppmat/94/suppmat_guo.tif. PMID- 15534870 TI - TNFalpha mediates sepsis-induced impairment of basal and leucine-stimulated signaling via S6K1 and eIF4E in cardiac muscle. AB - Decreased translation initiation adversely impacts protein synthesis and contributes to the myocardial dysfunction produced by sepsis. Therefore, the purpose of the present study was to identify sepsis-induced changes in signal transduction pathways known to regulate translation initiation in cardiac muscle and to determine whether the stimulatory effects of leucine can reverse the observed defects. To address this aim, sepsis was produced by cecal ligation and puncture (CLP) in anesthetized rats and the animals studied in the fasted condition 24 h later. Separate groups of septic and time-matched control rats also received an oral gavage of leucine. To identify potential mechanisms responsible for regulating cap-dependent mRNA translation in cardiac muscle, several eukaryotic initiation factors (eIFs) were examined. Under basal conditions, hearts from septic rats demonstrated a redistribution of the rate limiting factor eIF4E due to increased binding of the translational repressor 4E BP1 with eIF4E. However, this change was independent of an alteration in the phosphorylation state of 4E-BP1. The phosphorylation of mTOR, S6K1, the ribosomal protein (rp) S6, and eIF4G was not altered in hearts from septic rats under basal conditions. In control rats, leucine failed to alter eIF4E distribution but increased the phosphorylation of S6K1 and S6. In contrast, in hearts from septic rats leucine acutely reversed the alterations in eIF4E distribution. However, the ability of leucine to increase S6K1 and rpS6 phosphorylation in septic hearts was blunted. Sepsis increased the content of tumor necrosis factor (TNF)-alpha in heart and pre-treatment of rats with a TNF antagonist prevented the above mentioned sepsis-induced changes. These data indicate that oral administration of leucine acutely reverses sepsis-induced alterations eIF4E distribution observed under basal conditions but the anabolic actions of this amino acid on S6K1 and rpS6 phosphorylation remain blunted, providing evidence for a leucine resistance. Finally, TNFalpha, either directly or indirectly, appears to mediate the sepsis induced defects in myocardial translation initiation. PMID- 15534871 TI - Lysosomal and mitochondrial pathways in H2O2-induced apoptosis of alveolar type II cells. AB - Increasing evidence suggests a role for apoptosis in the maintenance of the alveolar epithelium under normal and pathological conditions. However, the signaling pathways modulating alveolar type II (AT II) cell apoptosis remain poorly defined. Here we investigated the role of lysosomes as modulators of oxidant-mediated AT II cell apoptosis using an in vitro model of H(2)O(2)-stress. H(2)O(2) stress led to time-dependent increases in intracellular oxidants, mitochondrial membrane polarization, cytochrome c release, lysosomal rupture, and AT II cells apoptosis. Increased apoptosis was prevented by specific inhibition of the caspase cascade using the broad-spectrum caspase inhibitor z-VAD-fmk or a caspase 3 inhibitor, or by using functional inhibitors for cathepsin D (pepstatin A) or cathepsin B. Inhibition of cathepsin D also prevented mitochondrial permeabilization and cythocrome c release suggesting that lysosomal rupture precedes and is necessary for the activation of the mitochondrial pathway of cell death. PMID- 15534872 TI - A review of current applications of mass spectrometry for biomarker/molecular tracer elucidation. AB - Mass spectrometry, especially coupled with gas chromatography or tandem, is the analytical method of choice in elucidation of biomarker compounds present in organic mixtures extracted from geological, environmental, or biological samples. This review describes the biomarker concept; i.e., the precursor natural products to the geological/environmental derivatives, and their application as tracers in the geosphere and ambient environment. The mass spectrometric methods currently utilized for such analyses are reviewed, and typical examples of applications are described with a general key to the literature. PMID- 15534873 TI - Mild phenotype in two unrelated patients with a partial deletion of 21q22.2-q22.3 defined by FISH and molecular studies. AB - We describe two unrelated patients with cytogenetically visible deletions of 21q22.2-q22.3 and mild phenotypes. Both patients presented minor dysmorphic features including thin marfanoid build, facial asymmetry, downward-slanting palpebral fissures, depressed nasal bridge, small nose with bulbous tip, and mild mental retardation (MR). FISH and molecular studies indicated common deleted areas but different breakpoints. In patient 1, the breakpoint was fine mapped to a 5.2 kb interval between exon 5 and exon 8 of the ETS2 gene. The subtelomeric FISH probe was absent on one homologue 21 indicating a terminal deletion spanning approximately 7.9 Mb in size. In patient 2, the proximal breakpoint was determined to be 300-700 kb distal to ETS2, and the distal breakpoint 2.5-0.3 Mb from the 21q telomere, indicating an interstitial deletion sized approximately 4.7-7.3 Mb. The 21q- syndrome is rare and typically associated with a severe phenotype, but different outcomes depending on the size and location of the deleted area have been reported. Our data show that monosomy 21q of the area distal to the ETS2 gene, representing the terminal 7.9 Mb of 21q, may result in mild phenotypes comprising facial anomalies, thin marfanoid build, and mild MR, with or without signs of holoprosencephaly. PMID- 15534874 TI - Multisystem study of 20 older adults with Williams syndrome. AB - To address the natural history of Williams syndrome (WS), we performed multisystem assessments on 20 adults with WS over 30 years of age and documented a high frequency of problems in multiple organ systems. The most striking and consistent findings were: abnormal body habitus; mild-moderate high frequency sensorineural hearing loss; cardiovascular disease and hypertension; gastrointestinal symptoms including diverticular disease; diabetes and abnormal glucose tolerance on standard oral glucose tolerance testing; subclinical hypothyroidism; decreased bone mineral density on DEXA scanning; and a high frequency of psychiatric symptoms, most notably anxiety, often requiring multimodal therapy. Review of brain MRI scans did not demonstrate consistent pathology. The adults in our cohort were not living independently and the vast majority were not competitively employed. Our preliminary findings raise concern about the occurrence of mild accelerated aging, which may additionally complicate the long-term natural history of older adults with WS. We provide monitoring guidelines to assist in the comprehensive care of adults with WS. PMID- 15534875 TI - Expression and characterization of a serine protease that preferentially cleaves insulin-like growth factor binding protein-5. AB - Insulin-like growth factor binding proteins (IGFBPs) play important roles in regulating the functions of insulin-like growth factors (IGFs). Because IGFBPs have very high affinity for IGF-I and IGF-II, they can regulate the amount of each growth factor that is able to bind to cell surface receptors, therefore, factors that alter IGFBP affinity have the capacity to regulate IGF actions. Protease activities that are present in cell culture systems and physiologic fluids have been shown to degrade IGFBP-5. Previously, a region of sequence in a serine protease was identified that was homologous with the N-terminal 90 amino acids of members of the IGFBP family and with members of the CCN family of proteins. In a prior study, the protease was expressed in human kidney cultured cells and the cell culture supernatants were shown to cleave IGFBP-5, however, it is unknown whether the purified protease would cleave IGFBP-5 and whether it would also cleave other specific forms of IGFBPs. In this study, we expressed this protease in an insect cell expression system, purified it to homogeneity and tested its capacity to cleave IGFBP-5. The expressed protease preferentially cleaved IGFBP-5, and it had minimal activity toward other forms of IGFBPs. The proteolytic activity of this IGFBPase is inhibited by serine protease inhibitors including PMSF and 3,4-dichloroisocoumarin, as well as by divalent metal ions such as, Zn and Cu. Mutation of the active site serine resulted in a major reduction in IGFBP-5 cleavage. The protease binds to heparin and its ability to degrade IGFBP-5 is blocked in the presence of heparin. Inhibition of the activity of the protease following its secretion by B104 cells resulted in inhibition of IGFBP-5 proteolysis and IGF-I stimulation of protein synthesis. Northern blotting revealed that the transcript was expressed in multiple human tissues, including placenta, uterus, prostate, testis, spinal cord, brain, liver, small intestine, thyroid, and spleen. The highest expression was in uterus and placenta, suggesting a possible role of sex steroids in regulating its expression. Understanding the mechanism of how cleavage of IGFBP-5 by this protease alters its activity will help to further our understanding of the biologic actions of the IGFs. PMID- 15534876 TI - High hydrostatic pressure inhibits the biosynthesis of eukaryotic elongation factor-2. AB - High continuous hydrostatic pressure is known to inhibit the total cellular protein synthesis. In this study, our goal was to identify pressure-regulated proteins by using two dimensional gel electrophoresis and mass spectrometry. This analysis showed that under 30 MPa continuous hydrostatic pressure the biosynthesis of eukaryotic elongation factor-2 (eEF-2) was inhibited both in HeLa carcinoma and T/C28a4 chondrocytic cell lines. Western blot analysis of HeLa cells revealed that the cellular protein level of eEF-2 decreased by 40%-50% within 12 h of the pressure treatment. However, the steady-state mRNA level of eEF-2 was not affected by the pressure. Cycloheximide addition after 4 h-pressure treatment suggested that the half-life of eEF-2 protein was shorter in pressurized cells. eEF-2 is responsible for the translocation of ribosome along the specific mRNA during translation, and its phosphorylation prevents the ribosomal translocation. Therefore, increased phosphorylation of eEF-2 was considered as one mechanism that could explain the reduced level of protein synthesis in pressurized HeLa cell cultures. However, Western blot analysis with an antibody recognizing the Thr56-phosphorylated form of eEF-2 showed that phosphorylation of eEF-2 was not elevated in pressurized samples. In conclusion, the inhibition of protein synthesis under high pressure occurs independent of the phosphorylation of eEF-2. However, this inhibition may result from the decrease of cellular eEF-2 protein. PMID- 15534877 TI - Exposure of human lymphocytes and lymphoblastoid cells to simulated microgravity strongly affects energy metabolism and DNA repair. AB - Exposure of freshly drawn lymphocytes and lymphoblastoid cells (LB and COR3) to simulated microgravity decreased the intracellular ATP concentration to 50%-40% of the value found in normal growth conditions. The decrease was reversible although recovery to normal values occurred only slowly both in lymphocytes and in lymphoblastoid cells. Poly(ADP-ribose) polymerase (PARP ) activity was increased indicating that cells exposed to conditions of reduced gravitation experience stress. Exposure to microgravity forces cells into a condition of metabolic quiescence in which they appear to be particularly sensitive to subsequent exposures to a genotoxic agent. Thus, treatment of cells with the strong redox agent potassium bromate under microgravity conditions, indicated an impairment in repair of DNA 8-hydroxy-2'-deoxyguanosine (8-OHdG), an oxidized derivative of deoxyguanosine. We conclude that gravitational modulation of the kind routinely obtained under laboratory conditions and during spaceflights is a stressful process to which cells appear to be extremely sensitive. These effects may reflect the physiological alterations observed in astronauts and in animals following spaceflights or exposure to conditions of simulated microgravity. PMID- 15534878 TI - La3+-promoted proliferation is interconnected with apoptosis in NIH 3T3 cells. AB - Lanthanum ion (La(3+)) has been reported to affect proliferation or apoptosis of different cells. In the present study, La(3+) was confirmed to promote both proliferation and apoptosis of NIH 3T3 cells at the same concentrations. La(3+) was shown to promote proliferation by helping the cells to pass through the G1/S restriction point and enter S phase, however, the proliferating cells induced by incubation with La(3+) eventually underwent apoptosis. The proliferation and apoptosis of NIH 3T3 cells induced by La(3+) were well correlated with cell cycle alterations. La(3+) caused the phosphorylation of extracellular signal-regulated kinase (ERK) 1/2; while inhibition of ERK phosphorylation by 2'-amino-3' methoxyflavone (PD98059) suppressed both proliferation and apoptosis induced by La(3+). Based on the above experimental results, we postulated that La(3+) promoted proliferation of NIH 3T3 cells could be interconnected with the cell apoptosis, possibly through cell cycle machinery. Our results thus support the recent hypothesis that proliferation and apoptosis of cell are intrinsically coordinated. PMID- 15534879 TI - beta(2)-glycoprotein I protects J774A.1 macrophages and human coronary artery smooth muscle cells against apoptosis. AB - beta(2)-Glycoprotein I (beta(2)-GPI) is a plasma glycoprotein with multifactorial relevance to clinical consequences. It was previously indicated that beta(2)-GPI can selectively bind to apoptotic cells. This study was designed to determine the role of beta(2)-GPI in apoptosis. Using an immunohistochemical study, we observed that beta(2)-GPI was co-localized with the apoptotic macrophages and smooth muscle cells (SMCs) of human coronary arteries. The contribution of beta(2)-GPI to apoptotic death was then investigated in vascular cells. Two nitric oxide (NO) donors, S-nitrosoglutathione (GSNO) and S-nitroso-N-acetyl penicillamine (SNAP) were used in this study to trigger apoptosis in J774A.1 macrophages and human coronary artery smooth muscle cells (HCASMC). Cell viability was significantly improved in beta(2)-GPI-treated cells. It was also possible to detect a remarkable inhibitory effect by beta(2)-GPI on the NO-induced apoptosis by preventing nuclear shrinkage. Furthermore, the NO-induced apoptosis was associated with increase in caspase-3 activity and in the protein levels of caspase-3, c-Fos, and c-Jun. However, all these apoptosis-related events were inhibited in vascular cells treated with 200 microg/ml beta(2)-GPI. This is the first study to show that beta(2)-GPI may be important in the prevention of apoptosis in vascular cells. PMID- 15534880 TI - Risk factors associated with earlier age of onset in familial pancreatic carcinoma. AB - BACKGROUND: An estimated 5-10% of all pancreatic adenocarcinomas have a hereditary association. The objective of the current study was to characterize the clinical and pathologic features of familial pancreatic carcinoma and to determine potential differences in demographics, risk factors, and outcomes between familial and sporadic pancreatic carcinoma populations. METHODS: A retrospective review was performed to identify patients diagnosed with pancreatic carcinoma who had an associated familial disposition. Demographic analyses and assessment of clinical features and treatment outcomes were performed for the familial subgroup, and the results were compared with observations made in the nonfamilial, or 'sporadic', population. RESULTS: Thirty of 826 patients (3.6%) had familial pancreatic carcinoma. Baseline demographics, resectability, and metastases were similar in both the familial cohort and the sporadic cohort. The mean age of onset was slightly lower in the familial cohort (57.6 years, compared with 61 years in the sporadic cohort). However, the familial population had a significantly greater proportion of patients who were diagnosed at age <50 years compared with the sporadic population (36.7% vs. 18.3%; P=0.017). A positive smoking history was more commonly associated with familial pancreatic carcinoma (87% vs. 66%; P=0.06). The overall median survival durations were 7 months and 6 months for the familial group and the sporadic group, respectively. CONCLUSIONS: Patients with familial pancreatic carcinoma present at an earlier age compared with their counterparts who have nonfamilial disease. Smoking may play a significant role in the risk or promotion of pancreatic carcinoma in patients with an inherited predisposition. PMID- 15534881 TI - Acute myeloid leukemia and Down syndrome evolution of modern therapy--state of the art review. AB - Over the past decade, a series of clinical reports have described the experience of Down syndrome (DS) children treated for acute myeloid leukemia (AML). Whereas prior to the first reports in the early 1980's it was felt that DS children with AML had poor outcomes, these clinical trials concluded that DS had a better outcome than non-DS (NDS) children with AML. With these recent reports, it is clear that DS children have a better outcome utilizing less intensive chemotherapy regimens. They also tolerate the more intensive regimens less well than the NDS children. This review focuses upon the six multi-institutional reports that described the DS and AML experience in order to better ascertain the chemotherapy combinations that may be useful in the future for these children. Regimens of varying intensity have all had similar outcomes. In general, the remission rates are approximately 90% with event-free survival (EFS) approximating 70-80%. Most recently, the clinical trials have been large enough to explore prognostic factors specifically in the DS children. This has identified that the younger DS children fair significantly better than the older children. AML in DS is unique and these differences in comparison to NDS children are highlighted. The significantly better outcomes for DS children likely represents a combination of the unique AML seen in DS children and the heightened sensitivity to cytarabine that DS AML cells have. Future trials should focus on age-stratified approaches that exploit the greater sensitivity of DS AML to cytarabine. PMID- 15534882 TI - Palliative radiation therapy for painful vertebral metastases: a practice survey. PMID- 15534883 TI - The SOD2 Val/Val genotype enhances the risk of nonsmall cell lung carcinoma by p53 and XRCC1 polymorphisms. AB - BACKGROUND: Exogenous reactive oxygen species (ROS) induces DNA damage. Manganese superoxide dismutase (SOD2) catalyzes the dismutation of superoxide radicals, a major type of ROS, into hydrogen peroxide. p53 is a tumor suppressor gene, and X ray cross-complementing group 1 (XRCC1) is involved in the base-excision repair of ROS-induced DNA damage. METHODS: The authors investigated whether the SOD2 Ala16Val polymorphism modifies the associations between p53 Arg72Pro and XRCC1 Arg399Gln polymorphisms and the risk of nonsmall cell lung carcinoma (NSCLC) in a case-control study of 935 Caucasian patients with NSCLC and 1233 healthy control participants. The results were analyzed using logistic regression models that were adjusted for possible confounding variables. RESULTS: There was no association between p53 or XRCC1 polymorphism and NSCLC risk for individuals with SOD2 Ala/Ala or Ala/Val genotype. For individuals with the SOD2 Val/Val genotype, greater risks were found in association with p53 (variant Pro allele vs. Arg/Arg), XRCC1 (variant Gln allele vs. Arg/Arg), and the combination of the two polymorphisms ("double variant" vs. "double wild type"), with the adjusted odds ratios (ORs) of 1.84 (95% confidence interval [95% CI], 1.20-2.82), 1.39 (95% CI, 0.98-2.21), and 2.54 (95% CI, 1.38-4.68), respectively. Furthermore, the greater risk for the double variant of p53 and XRCC1 in the SOD2 Val/Val genotype group was specific only for patients with adenocarcinoma and not for patients with squamous cell carcinoma, with adjusted ORs of 3.31 (95% CI, 1.68-6.51) and 0.69 (95% CI, 0.24-2.02), respectively. CONCLUSIONS: The SOD2 Val/Val genotype may increase the risk of NSCLC carried by XRCC1 and p53 polymorphisms, particularly for adenocarcinoma. PMID- 15534884 TI - Successful human ovarian autotransplantation to the upper arm. AB - In young patients with cervical carcinoma, the standard surgical treatment is often followed by postoperative radiotherapy. This treatment strategy, although resulting in a significant increase in cure rates, often causes infertility and premature ovarian failure. Ovarian autotransplantation outside the field of radiotherapy is a new technique to preserve gonadal function. One ovary was transplanted to the left upper arm during the surgical treatment for cervical carcinoma. Vascular anastomoses were performed by microsurgery. After transplantation, the ovary showed adequate arterial and venous blood flow. By clinical examination and ultrasound monitoring, the ovarian cycles remained regular and follicle growth occurred normally for > 1 year. Heterotopic autotransplantation of the ovary to the upper arm is a promising surgical technique to maintain the ovarian function in women who become menopausal due to cancer treatment regimens. PMID- 15534885 TI - Understanding of an aggregate probability statement by patients who are offered participation in Phase I clinical trials. AB - BACKGROUND: There is concern that patients with poor numeracy may have difficulty understanding the information necessary to make informed treatment decisions. The authors sought to characterize a special form of numeracy among patients with advanced cancer who were offered participation in Phase I oncology clinical trials. METHODS: Surveys were administered to 328 cancer patients who were considering Phase I trials. Their frequency-type numeracy was assessed using a multiple-choice question involving a hypothetical scenario in which a physician stated that an experimental treatment would control cancer in "40% of cases like yours." In univariate and multivariable analyses, patient characteristics that were associated with better numeracy were identified. RESULTS: The correct frequency-type interpretation was selected by 72% of respondents. Fourteen percent of respondents incorrectly selected a belief-type answer, "The doctor is 40% confident that the treatment will control my cancer." In a multivariable model, patients who answered incorrectly tended to have less formal education and less experience with experimental therapies. CONCLUSIONS: Because the misunderstandings some patients demonstrated may influence their treatment decision making adversely, it is critical to identify such patients and to give them special consideration when communicating information about potential risks and benefits of treatment. PMID- 15534886 TI - Comparison of stratification and adaptive methods for treatment allocation in an acute stroke clinical trial by Christopher J. Weir and Kennedy R. Lees, Statistics in Medicine 2003; 22:705-726. PMID- 15534888 TI - Case-sibling gene-association studies for diseases with variable age at onset. AB - Studies which compare cases to disease-free siblings are useful for assessing association between a genetic locus and a phenotypic trait, as they eliminate the possibility of confounding by population stratification. Many analytic methods for such family-based studies are based on a binary disease model. However, complex diseases have variable age at onset. Consequently, binary-outcome methods can be inefficient or biased. We review methods for analysing censored age-at onset data from family studies, including stratified Cox regression and genotype decomposition regression, an unstratified procedure which regresses age-at-onset on between- and within-family genotype components. We also introduce a retrospective likelihood for censored age-at-onset data, which requires an external estimate of the baseline hazard. Stratified Cox regression does not use controls who have not attained the age of their case sibling(s), potentially leading to a loss of efficiency. Both genotype-decomposition regression and the retrospective likelihood use these younger controls. We assess the performance of these methods via simulation studies. Stratified Cox regression and the retrospective likelihood have appropriate type I error rates in almost all situations studied; genotype-decomposition regression is often anti-conservative. Away from the null, confidence intervals for the relative risk derived from stratified Cox regression are anti-conservative when the disease is rare and case rich families are sampled. The retrospective likelihood is more efficient than stratified Cox regression and its confidence intervals have correct coverage when the disease is rare or the estimate of the baseline hazard is reasonably accurate. These results suggest that when estimating genotype relative risks is the principal analytic goal, stratified Cox regression is appropriate as long as the disease is common; when the disease is rare, the retrospective likelihood may be more appropriate. PMID- 15534890 TI - Box-Cox transformation of left-censored data with application to the analysis of coronary artery calcification and pharmacokinetic data. AB - Box-Cox transformation is investigated for regression models for left-censored data. Examples are provided using coronary calcification data from the Multi Ethnic Study of Atherosclerosis and pharmacokinetic data of a nicotine nasal spray. PMID- 15534891 TI - Evaluation of log-rank tests for infrequent observations from a multi-state process, with application to HPV vaccine efficacy. AB - Genital infection by human papillomavirus (HPV) is a common sexually transmitted disease, with over 25 per cent prevalence among young women in the US. Infections are usually without symptoms and transient (or reversible), but a small proportion of infections persist and are believed to be responsible for nearly all cervical cancers and precursor lesions such as cervical intraepithelial neoplasia (CIN). Therefore, successful vaccines against persistent HPV infections could have a great impact in preventing cervical cancers. In trials being planned, ongoing, and recently completed, a log-rank or a similar test may be employed to assess a vaccine effect in comparison to placebo, with an infection 'event' defined to capture persistent but not transient infections. However, it is not clear how best to define such an event, because (1) diagnostic tests cannot distinguish a persistent from a transient infection, (2) participants are only examined periodically, and (3) there can be misclassification errors in the detection of infections. This paper evaluates several definitions of persistent infection that are based on periodically observed infection statuses by postulating a multi-state model for persistent and transient infections. The type I error and the power of tests on vaccine efficacy based on these operational definitions are then examined under various scenarios of how a vaccine might affect the infection-disease process. We find that none of the candidates performs satisfactorily, thus raising concerns that clinical trials based only on infection endpoints will not be reliable. PMID- 15534892 TI - Modelling neuromotor ratings with floor-effects. AB - Associated movements (AMs) are a classical diagnostic tool to assess differences between normal children and children with some motor dysfunction. This paper presents a methodology to produce age- and gender-dependent reference-curves for AMs of normal children, for various tasks of a test battery. Data available consist of separate ratings of duration and extent of AMs, which are ordinal quantities with few levels. Other problems are severe age- and gender-dependent floor-effects (as well as some ceiling-effects), leaving little information for analysis at older ages. To get a better scale, we combined the two ordinal ratings into one meaningful and quasi-continuous quantity referred to as intensity of AMs. In order to solve problems due to floor-effects, ceiling effects and discreteness, we assumed left- , right- and interval-censored values, respectively. We considered a censored regression problem and postulated a truncated normal distribution for the non-censored values (after an appropriate transformation of the data). Using Wei and Tanner's poor man's data augmentation algorithm, together with the technique of linear mixed effects modelling, useful reference-curves could be produced. In contrast to the cumulative probabilities approach for ordinal data, our methodology allows the calculation of individual age- and gender-standardized values, which puts us in a position to investigate numerous scientific questions. PMID- 15534893 TI - A Welch-type test for homogeneity of contrasts under heteroscedasticity with application to meta-analysis. AB - A common problem that arises in the meta-analysis of several studies, each with independent treatment and control groups, is to test for the homogeneity of effect sizes without the assumptions of equal variances of the treatment and the control groups and of equal variances among the separate studies. A commonly used test statistic, frequently denoted as Q, is the weighted sum of squares of the differences of the individual effect sizes from the mean effect size, with weights inversely proportional to the variances of the effect sizes. The primary contributions of this article are the presentation of improved and very accurate approximations to the distributions of the Q statistic when the effect size is a linear contrast such as the difference between the treatment and control means. Our improved approximation to the distribution of Q under the null hypothesis is based on a multiple of an F-distribution; its use yields a substantial reduction in the type I error rate of the homogeneity test. Our improved approximation to the distribution of Q under an alternative hypothesis is based on a shift of a chi-square distribution; its use allows for much greater accuracy in the computation of the power of the homogeneity test. These two improved approximate distributions are developed using the Welch methodology of approximating the moments of Q by the use of multivariate Taylor expansions. The quality of these approximations is studied by simulation. A secondary contribution of this article is a study of how best to combine the variances of the treatment and control groups (needed for the calculation of weights in the Q statistic). Our conclusion, based on simulations, is that use of pooled variances can result in substantially erroneous conclusions. PMID- 15534894 TI - Analysis of clustered and interval censored data from a community-based study in asthma. AB - Many authors in recent years have proposed extensions of familiar survival analysis methodologies to apply in dependent data settings, for example, when data are clustered or subject to repeated measures. However, these extensions have been considered largely in the context of right censored data. In this paper, we discuss a parametric frailty model for the analysis of clustered and interval censored failure time data. Details are presented for the specific case where the underlying time to event data follow a Weibull distribution. Maximum likelihood estimates will be obtained using commercially available software and the empirical efficiency of these estimators will be explored via a simulation study. We also discuss a score test to make inferences about the magnitude and significance of over-dispersion in clustered data settings. These methods will be illustrated using data from the East Boston Asthma Study. PMID- 15534895 TI - Likelihood-based modelling of age-related normal ranges for ordinal measurements: changes in visual acuity through early childhood. AB - This paper investigates the construction of age-related standards for ordinal outcome data. Asymmetric logistic models are used to describe the age-related changes in the cumulative probabilities for each ordinal level. Maximum likelihood estimation of model parameters allows the use of likelihood ratio tests to ascertain the appropriate model complexity. In contrast to methodologies for constructing age-related standards where the outcome is continuous, we show how the methodology leads directly to centile estimation for individuals. The method is illustrated using visual acuity measurements collected from 2968 children between 2 and 9 years of age made on a 30-point ordinal scale. We show how, in this instance, smoothing of parameters across ordinal categories leads to reduced validity of the centiles, justifying the need for specialized methodology for non-continuous outcomes. PMID- 15534896 TI - Score tests for dose effect in the presence of non-responders. AB - When only a certain proportion of subjects respond to treatment ('responders') or may never experience an event of interest (thus 'cured'), mixture models often lead to increased understanding of the treatment or disease process. This paper focuses on hypothesis testing in a dose-response framework and shows that increased power is possible by using a mixture model where both the logit of the response rate and the response mean are linear functions of the dose level. Three score tests are developed for testing an overall effect and permutation methods are used to control the type I error. Extensive simulations establish the power properties of the tests and show that our proposed score test has the best performance. The approach is illustrated by a multi-country clinical trial of rapid acting Intramuscular Olanzapine. PMID- 15534897 TI - On simultaneous assessment of sensitivity and specificity when combining two diagnostic tests. AB - Diagnostic tests are seldom adopted in isolation. Few tests have high sensitivity and specificity simultaneously. In these cases, one can increase either the sensitivity or the specificity by combining two component tests under either the 'either positive' rule or the 'both positive' rule. However, there is a tradeoff between sensitivity and specificity when these rules are applied. We propose three statistical procedures to simultaneously assess the sensitivity and specificity when combining two component tests. Measurements of interest include rate difference and rate ratio. Our empirical results demonstrate that (i) the asymptotic test procedures for both measurements and approximate test procedure for rate difference possess inflated type I error rate; (ii) the exact test procedures for both measurements possess deflated type I error rate; and (iii) the approximate (unconditional) test procedure for rate ratio becomes an reliable alternative and nicely controls the actual type I error rate in small to moderate sample sizes. Moreover, the approximate (unconditional) test procedure is computationally less intensive than the exact (unconditional) test procedure. We illustrate our methodologies with a real example from a residual nasopharyngeal carcinoma (RNP) study. PMID- 15534898 TI - Comment on: Estimate of standard deviation for a log-transformed variable using arithmetic means and standard deviations by Quan Hui and Zhang Ji, Statistics in Medicine 2003; 22: 2723-2736. PMID- 15534899 TI - Estimating treatment effect in the presence of non-compliance measured with error: precision and robustness of data analysis methods. AB - Non-compliance with the nominal prescribed dosage causes unintended variability in actual drug exposure during clinical trials. In the ideal case that compliance is not a confounder, and it is known--hence actual dosage is known--true dose response can be validly estimated. Measuring compliance presents a challenge, however. A simulation study of the case that dosage history questionnaires (C(Q)- usually over-optimistic estimates of actual compliance) are available in all subjects enrolled in a clinical trial, but accurate compliance measurements (C- e.g. from electronic medication event monitors), are only available in a (random) fraction of subjects is reported. It reveals that a 'Maximum Penalized Marginal Likelihood' (MPML) method which uses all compliance data, effectively calibrating C(Q) to C, is superior to other methods which use only one compliance measure, or both, or neither (neither = ITT, intention to treat, which assumes actual dosage equals nominal dosage), but do not calibrate. MPML yields the most precise estimates of dose-response over widely varying clinical trial designs, extremes in quality and quantity of compliance information, and a range of drug effect sizes. It is most beneficial when compliance data are sparse and maintains good performance even when its key assumptions are somewhat violated. PMID- 15534901 TI - Evaluation of an antibiotic intravenous to oral sequential therapy program. AB - AIM: This study was designed to analyse the drug consumption difference and economic impact of an antibiotic sequential therapy focused on quinolones. METHOD: We studied the consumption of quinolones (ofloxacin/levofloxacin and ciprofloxacin) 6 months before and after the implementation of a sequential therapy program in hospitalised patients. It was calculated for each antibiotic, in its oral and intravenous forms, in defined daily dose (DDD/100 stays per day) and economical terms (drug acquisition cost). At the beginning of the program ofloxacin was replaced by levofloxacin and, since their clinical uses are similar, the consumption of both drugs was compared during the period. RESULTS: In economic terms, the consumption of intravenous quinolones decreased 60% whereas the consumption of oral quinolones increased 66%. In DDD/100 stays per day, intravenous forms consumption decreased 53% and oral forms consumption increased 36%. CONCLUSIONS: Focusing on quinolones, the implementation of a sequential therapy program based on promoting an early switch from intravenous to oral regimen has proved its capacity to alter the utilisation profile of these antibiotics. The program has permitted the hospital a global saving of 41420 dollars for these drugs during the period of time considered. PMID- 15534902 TI - Determinants of medicine use in a Swedish primary health care practice population. AB - BACKGROUND: Pharmaceuticals represent an increasing share of private and public health care expenditures. The aim of this study was to characterise users and to determine the pattern of uses of prescribed medicines and to identify determinants of medicine use in a multi-ethnic Swedish general practice population. METHODS: The study was performed in 1055 of 1442 consecutive adult patients visiting the Jordbro Health Centre (JHC) in Stockholm, Sweden. RESULTS: In a regression analysis adjusted for the influence of confounders, subjects reporting 10-30 complaint symptoms, subjects with chronic disease and subjects that had a cost limitation card all reported a higher number of used medicines than subjects with a few (0-10) symptoms and those without chronic disease and a cost limitation card. In a Poisson's regression analysis, high age, female gender, not working, more than 10 symptoms, chronic disease and having a cost limitation card were related to use of prescribed medicines. All included variables predicted 19% of the explanation. Marital status, multi-symptomatology, chronic disease and having a cost limitation card were related to polypharmacy in a logistic regression analysis. CONCLUSION: A high number of complaint symptoms, chronic disease and having a cost limitation card were all significantly and independently related to use of medicines and polypharmacy. PMID- 15534903 TI - Hepatocellular carcinoma treated with interventional procedures: CT and MRI follow-up. AB - In the past decade, a variety of interventional procedures have been employed for local control of hepatocellular carcinoma (HCC). These include transcather arterial chemoembolization (TACE) and several tumour ablation techniques, such as percutaneous ethanol injection (PEI), radio-frequency ablation (RFA), or percutaneous microwave coagulation therapy (PMC), laser-induced interstitial thermotherapy (LITT), etc. For a definite assessment of the therapeutic efficacy of interventional procedures, histological examination using percutaneous needle biopsy may be the most definite assessment of the therapeutic efficacy of interventional therapy, however, it is invasive and the specimen retrieved does not always represent the entire lesion owing to sampling errors. Therefore, computed tomography (CT) and magnetic resonance imaging (MRI) play a crucial role in follow-up of HCC treated by interventional procedures, by which the local treatment efficacy, recurrent disease and some of therapy-induced complications are evaluated. Contrast enhanced axial imaging (CT or MR imaging) may be the most sensitive test for assessing the therapeutic efficacy. The goal of the review was to describe the value of CT and MRI in the evaluation of interventional treatments. PMID- 15534904 TI - Lymph node metastasis in early gastric cancer with submucosal invasion: feasibility of minimally invasive surgery. AB - AIM: To explore the feasibility of performing minimally invasive surgery (MIS) on subsets of submucosal gastric cancers that are unlikely to have regional lymph node metastasis. METHODS: A total of 105 patients underwent radical gastrectomy with lymph node dissection for submucosal gastric cancer at our hospital from January 1995 to December 1995. Besides investigating many clinicopathological features such as tumor size, gross appearance, and differentiation, we measured the depth of invasion into submucosa minutely and analyzed the clinicopathologic features of these patients regarding lymph node metastasis. RESULTS: The rate of lymph node metastasis in cases where the depth of invasion was <500 microm, 500 2000 microm, or >2000 microm was 9% (2/23), 19% (7/36), and 33% (15/46), respectively (P<0.05). In univariate analysis, no significant correlation was found between lymph node metastasis and clinicopathological characteristics such as age, sex, tumor location, gross appearance, tumor differentiation, Lauren's classification, and lymphatic invasion. In multivariate analysis, tumor size (>4 cm vs <=2 cm, odds ratio = 4.80, P = 0.04) and depth of invasion (>2000 microm vs <=500 microm, odds ratio = 6.81, P = 0.02) were significantly correlated with lymph node metastasis. Combining the depth and size in cases where the depth of invasion was less than 500 microm, we found that lymph node metastasis occurred where the tumor size was greater than 4 cm. In cases where the tumor size was less than 2 cm, lymph node metastasis was found only where the depth of tumor invasion was more than 2000 microm. CONCLUSION: MIS can be applied to submucosal gastric cancer that is less than 2 cm in size and 500 microm in depth. PMID- 15534905 TI - Microarray-based method for detecting methylation changes of p16(Ink4a) gene 5' CpG islands in gastric carcinomas. AB - AIM: Aberrant DNA methylation of CpG site is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation of the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. However, large-scale analysis of candidate genes has so far been hampered by the lack of high-throughput approach for analyzing DNA methylation. The aim of this study was to describe a microarray-based method for detecting changes of DNA methylation in cancer. METHODS: This method used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Therefore, the amplified product might contain a pool of DNA fragments with altered nucleotide sequences due to differential methylation status. Nine sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of p16 gene CpG islands in gastric carcinomas. The results were further validated by methylation-specific PCR (MSP). RESULTS: The experimental results showed that the microarray assay could successfully detect methylation changes of p16 gene in 18 gastric tumor samples. Moreover, it could also potentially increase the frequency of detecting p16 methylation from tumor samples than MSP. CONCLUSION: Microarray assay could be applied as a useful tool for mapping methylation changes in multiple CpG loci and for generating epigenetic profiles in cancer. PMID- 15534906 TI - Alteration of p53 and p21 during hepatocarcinogenesis in tree shrews. AB - AIM: To investigate p53 mutation and p21 expression in hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B(1) (AFB(1)) in tree shrews, and to reveal the role of these genes in hepatocarcinogenesis. METHODS: Tree shrews were divided into four groups: group A, those infected with HBV and fed with AFB(1) (n = 39); group B, those infected with HBV alone (n = 28); group C, those fed with AFB(1) alone (n = 29); and group D, normal controls (n = 20). The tree shrews underwent liver biopsies once every 15 wk. Expression of p53 and p21 proteins and genes in the biopsies and tumor tissues of the experimental tree shrews was detected, respectively, by immunohistochemistry, and by Southern blotting and reverse transcription-polymerase chain reaction and sequencing. RESULTS: The incidence of hepatocellular carcinomas (HCC) was higher in group A (66.7%) than that in group B (3.57%) and C (30%). The time of HCC occurrence was also earlier in group A than that in group C (120.0+/-16.6 wk vs 153.3+/-5.8 wk, respectively, P<0.01). p53 protein was not detected by immunohistochemistry in all groups before the 75(th) wk of the experiment. At the 105(th) wk, the positive rates fo p53 were 78.6%, 60% and 71.4% in groups A, B and C, respectively, which were significantly higher than that in group D (10%) (all P<0.05). An abnormal band of p53 gene was observed in groups A and C. The mutation points of p53 gene in tree shrews with HCC were at codons 275, 78 and 13. The nucleotide sequence and amino acid sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homologies with those of human p53, respectively. The immunopositivity for p21 was found before HCC development. The incidence of HCC was significantly higher in tree shrews that were positive for p21 than those negative for p21 (80.0% vs 11.0%, P<0.001). The incidence of HCC in p21 positive animals in group A was significantly higher than those positive for p21 in group C (P<0.05). CONCLUSION: A remarkable synergistic effect on HCC development exists between HBV and AFB(1). p53 mutation promotes the development of HCC. HBV and AFB(1) may synergistically induce p53 gene mutation, and stimulate ras gene expression. ras gene is activated at the earlier stage during hepatocarcinogenesis. p21 protein may be an early marker, and the alterations of p53 may be a late event in the development of HCC. PMID- 15534907 TI - A novel, rapid strategy to form dendritomas from human dendritic cells and hepatocellular carcinoma cell line HCCLM3 cells using mature dendritic cells derived from human peripheral blood CD14+ monocytes within 48 hours of in vitro culture. AB - AIM: Dendritomas formed by fusing cancer cells to dendritic cells have already been applied to clinical treatment trial of several types of cancers. Dendritic cells for the fusion in most trials and experiments were from blood monocytes in standard 7-d protocol culture, which requires 5-7 d of culture with granulocyte macrophage-colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4), followed by 2-3 d of activation with a combination of proinflammatory mediators such as tumor necrosis factoralpha (TNFalpha), interleukin-1beta (IL-1beta), interleukin 6 (IL-6) and prostaglandin E(2) (PGE(2)). One study showed that mature monocyte derived dendritic cells could be obtained within 48 h of in vitro culture with the same protocol as standard 7-d culture and referred to as FastDCs. Here we aimed to fuse human hepatocellular carcinoma cell line HCCLM3 cells with mature monocyte-derived dendritic cells within 48 h of in vitro culture (FastDC). METHODS: HCCLM3 cells were cultured in RPMI 1640 with 150 mL/L fetal calf serum (FCS). CD14+monocytes from healthy human peripheral blood were purified with MACS CD14 isolation kit and cultured in six-well plates in fresh complete DC medium containing RPMI-1640, 20 mL/L heat inactivated human AB serum, 2 mmol/L L glutamine, 100 microg/mL gentamicin, 1 000 U/mL GM-CSF and 500 U/mL IL-4 for 24 h, then proinflammatory mediators such as TNFalpha (1 000 U/mL), IL-1beta (10 ng/mL), IL-6 (10 ng/mL) and PGE(2) (1 microg/mL) were supplemented for another 24 h, and thus mature FastDCs were generated. HCCLM3 cells and FastDCs were labeled with red fluorescent dye PKH26-GL and green fluorescent dye PKH67-GL respectively. After the red fluorescent-stained HCCLM3 cells were irradiated with 50 Gy, FastDCs and irradiated HCCLM3 cells were fused in 500 mL/L polyethylene glycol(PEG)+100 mL/L dimethyl sulfoxide (DMSO) to generate novel dendritomas. The FastDCs and novel dendritomas were immunostained with anti-CD80, anti-CD86, anti CD83, anti-HLA-DR mAbs and analyzed by fluorescence-activated cell sorting (FACS). Novel dendritomas were nucleus-stained with Hoechst 33258 and analyzed by confocal laser scanning microscopy. RESULTS: Mature FastDCs with highly expressed surface markers CD80, CD86, CD83 and HLA-DR were generated within 48 h in vitro. Novel dendritomas with dual red-green fluorescence were constructed fast and successfully, and FACS analysis showed that the fusion efficiency was 24.27% and the novel dendritomas expressed the same activation markers as FastDCs. Confocal laser scanning microscopy analysis showed representative images of dendritomas. CONCLUSION: Dendritomas can be formed fast with mature FastDCs from healthy human peripheral blood monocytes (PBMC) by incubation with GM-CSF and IL-4 for 24 h and by activation with proinflammatory mediators for an additional period of 24 h. Owing to shorter time required for in vitro DCs development, the generation of these novel dendritomas reduced labor and cost. This rapid method for formation of dendritomas may represent a new strategy for immunotherapy of hepatocellular carcinoma. PMID- 15534908 TI - Differentially expressed genes between solitary large hepatocellular carcinoma and nodular hepatocellular carcinoma. AB - AIM: To study the difference in gene expression between solitary large hepatocellular carcinoma (SLHCC) and nodular hepatocellular carcinoma (NHCC). METHODS: Polymerase chain reaction (PCR) products of 8464 human genes were spotted on a chip in array. DNAs were then fixed on a glass plate. Total RNA was isolated from freshly excised human SLHCC (n = 7) and NHCC (n = 15) tissues, and was reversely transcribed to cDNAs with the incorporation of fluorescent dUTP for preparation of hybridization probes. The mixed probes were then hybridized to the cDNA microarray. After highly stringent washing, cDNA microarray was scanned for the fluorescent signals to display the difference between the two kinds of HCC. In addition, the expression of RhoC and protocadherin LKC was also detected with the reverse transcriptase polymerase chain reaction (RT-PCR) method. RESULTS: Among the 8464 human genes, 668 (7.89%) genes were expressed differentially at the mRNA levels between SLHCC and NHCC. Three hundred and fifty five (4.19%) genes, including protocadherin LKC, were up-regulated, whereas 313 (3.70%) genes, including RhoC, were down-regulated. The mRNA expression levels of RhoC and protocadherin LKC were confirmed by RT-PCR. Analysis of differentially expressed genes confirmed that our molecular data obtained by cDNA microarray were consistent with the published biochemical and clinical observations of SLHCC and NHCC. CONCLUSION: cDNA microarray is an effective technique in screening the difference in gene expression between SLHCC and NHCC. Many of these differentially expressed genes are involved in the invasion and metastasis of HCC. Further analysis of these genes will help to understand the different molecular mechanisms of SLHCC and NHCC. PMID- 15534909 TI - End-of-treatment virologic response does not predict relapse after lamivudine treatment for chronic hepatitis B. AB - AIM: Attaining hepatitis B e antigen (HBeAg) seroconversion during lamivudine treatment is associated with fewer relapses in HBeAg-positive patients. In HBeAg negative patients, predictors for post-treatment relapse remain largely unknown. We therefore studied whether end-of-treatment virologic response correlated with relapse after lamivudine treatment. METHODS: We prospectively analyzed 12 HBeAg negative patients and 14 HBeAg-positive patients with chronic hepatitis B, who received at least 9 mo of lamivudine treatment and were followed up for 12 mo post-treatment. Relapse of hepatitis B activity was defined by an elevation of serum ALT level above twice the upper limit of normal as well as reappearance of serum HBV DNA by the branched DNA assay or HBeAg during the follow-up period. The serum viral loads during and at the end of treatment were further determined by a quantitative real-time polymerase chain reaction assay. RESULTS: Relapse occurred in 6 (50.0%) HBeAg-negative patients within 12 mo post-treatment. Two relapsers had end-of-treatment serum viral load <1 000 copies/mL, the proportion was not significantly different from that in the 6 non-relapsers (33.3% vs 16.7%; P = 1.00). Hepatitis B virus (HBV) DNA levels did not correlate with post-treatment relapse in HBeAg-positive patients either. However, genotype C patients tended to have a lower relapse rate than genotype B patients (14.3% vs 57.9%, P = 0.08). CONCLUSION: Our results suggest that end-of-treatment virologic response cannot predict post-treatment relapse in patients with HBeAg-negative or -positive chronic hepatitis B. The impact of HBV genotype on the response to lamivudine treatment awaits further studies. PMID- 15534910 TI - Forecasting model for the incidence of hepatitis A based on artificial neural network. AB - AIM: To study the application of artificial neural network (ANN) in forecasting the incidence of hepatitis A, which had an autoregression phenomenon. METHODS: The data of the incidence of hepatitis A in Liaoning Province from 1981 to 2001 were obtained from Liaoning Disease Control and Prevention Center. We used the autoregressive integrated moving average (ARIMA) model of time series analysis to determine whether there was any autoregression phenomenon in the data. Then the data of the incidence were switched into [0,1] intervals as the network theoretical output. The data from 1981 to 1997 were used as the training and verifying sets and the data from 1998 to 2001 were made up into the test set. STATISTICA neural network (ST NN) was used to construct, train and simulate the artificial neural network. RESULTS: Twenty-four networks were tested and seven were retained. The best network we found had excellent performance, its regression ratio was 0.73, and its correlation was 0.69. There were 2 input variables in the network, one was AR(1), and the other was time. The number of units in hidden layer was 3. In ARIMA time series analysis results, the best model was first order autoregression without difference and smoothness. The total sum square error of the ANN model was 9 090.21, the sum square error of the training set and testing set was 8 377.52 and 712.69, respectively, they were all less than that of ARIMA model. The corresponding value of ARIMA was 12 291.79, 8 944.95 and 3 346.84, respectively. The correlation coefficient of nonlinear regression (R(NL)) of ANN was 0.71, while the R(NL) of ARIMA linear autoregression model was 0.66. CONCLUSION: ANN is superior to conventional methods in forecasting the incidence of hepatitis A which has an autoregression phenomenon. PMID- 15534911 TI - The effect of adenovirus expressing wild-type p53 on 5-fluorouracil chemosensitivity is related to p53 status in pancreatic cancer cell lines. AB - AIM: There are conflicting data about p53 function on cellular sensitivity to the cytotoxic action of 5-fluorouracil (5-FU). Therefore the objective of this study was to determine the combined effects of adenovirus-mediated wild-type (wt) p53 gene transfer and 5-FU chemotherapy on pancreatic cancer cells with different p53 gene status. METHODS: Human pancreatic cancer cell lines Capan-1(p53mut), Capan 2(p53wt), FAMPAC(p53mut), PANC1(p53mut), and rat pancreatic cancer cell lines AS(p53wt) and DSL6A(p53null) were used for in vitro studies. Following infection with different ratios of Ad-p53-particles (MOI) in combination with 5-FU, proliferation of tumor cells and apoptosis were quantified by cell proliferation assay (WST-1) and FACS (PI-staining). In addition, DSL6A syngeneic pancreatic tumor cells were inoculated subcutaneously in to Lewis rats for in vivo studies. Tumor size, apoptosis (TUNEL) and survival were determined. RESULTS: Ad-p53 gene transfer combined with 5-FU significantly inhibited tumor cell proliferation and substantially enhanced apoptosis in all four cell lines with an alteration in the p53 gene compared to those two cell lines containing wt-p53. In vivo experiments showed the most effective tumor regression in animals treated with Ad-p53 plus 5 FU. Both in vitro and in vivo analyses revealed that a sublethal dose of Ad-p53 augmented the apoptotic response induced by 5-FU. CONCLUSION: Our results suggest that Ad-p53 may synergistically enhance 5-FU-chemosensitivity most strikingly in pancreatic cancer cells lacking p53 function. These findings illustrate that the anticancer efficacy of this combination treatment is dependent on the p53 gene status of the target tumor cells. PMID- 15534912 TI - Enhanced anti-apoptosis and gut epithelium protection function of acidic fibroblast growth factor after cancelling of its mitogenic activity. AB - AIM: Mitogenic and non-mitogenic activities of fibroblast growth factor (FGF) are coupled to a range of biological functions, from cell proliferation and differentiation to the onset of many diseases. Recent reports have shown that acidic fibroblast growth factor (aFGF) has a powerful anti-apoptosis function, which may have potentially therapeutical effect on gut ischemia and reperfusion injuries. However, whether this function depends on its mitogenic or non mitogenic activity remains unclear. In this study, we identified the source of its anti-apoptosis function with a mutant, aFGF28-154 and observed its effect on reducing gut ischemia and reperfusion injury. METHODS: aFGF28-154 was generated by amplification of appropriate DNA fragments followed by subcloning the products into pET-3c vectors, then they were expressed in BL21 (DE3) cells and purified on an M2 agarose affinity column. This mutant aFGF28-154 maintained its non mitogenic activity and lost its mitogenic activity. With a dexamethasone (DEX) induced mouse thymocyte apoptosis model in vitro and in vivo, we studied the anti apoptotic function of aFGF28-154. Also, in vivo study was performed to further confirm whether aFGF28-154 could significantly reduce apoptosis in gut epithelium after gut ischemia-reperfusion injury in rats. Based on these studies, the possible signal transduction pathways involved were studied. RESULTS: With a dexamethasone (DEX)-induced mouse thymocyte apoptosis model in vitro and in vivo, we found that the anti-apoptotic function of aFGF28-154 was significantly enhanced when compared with the wild type aFGF. In vivo study further confirmed that aFGF28-154 significantly reduced apoptosis in gut epithelium after gut ischemia-reperfusion injury in rats. The mechanisms of anti-apoptosis function of aFGF28-154 did not depend on its mitogenic activity and were mainly associated with its non-mitogenic activities, including the intracellular calcium ion balance protection, ERK1/2 activation sustaining and cell cycle balance. CONCLUSION: These findings emphasize the importance of non-mitogenic effects of aFGF, and have implications for its therapeutic use in preventing apoptosis and other injuries in tissues and internal organs triggered by ischemia-reperfusion injury. PMID- 15534913 TI - Genetic alterations and reduced expression of tumor suppressor p33(ING1b) in human exocrine pancreatic carcinoma. AB - AIM: To detect the expression of p33(ING1b) protein and the change of p33(ING1b) gene in pancreatic carcinoma and to evaluate the significance of p33(ING1b) in pancreatic cell carcinogenesis. METHODS: Pathological specimens from pancreatic carcinoma and matched non-tumor pancreatic tissues were examined for p33(ING1b) expression and mutation by immunohistochemistry, polymerase chain reaction single strand conformation polymorphisms (PCR-SSCP) and loss of heterozygosity (LOH). RESULTS: The rate of p33(ING1b) protein expression was 85% (34/40). A single germline missense mutation was detected in 1 of 40 tumors located at codon 215:TGC-TCC (Cys-Ser). Fourteen (60.9%) of 23 tumor samples showed LOH in all of the informative markers tested, but no mutation was detected in these tumors and only two of the informative tumors lacked expressions of p33(ING1b) protein. CONCLUSION: Mutation and loss of expression are not the main reasons for the disfunction of p33(ING1b) in pancreatic carcinoma, an abnormality at the level of chromosome and/or transcription may inhibit their normal functions, potentially contributing to pancreatic cell carcinogenesis. PMID- 15534914 TI - High-yield expression of recombinant SARS coronavirus nucleocapsid protein in methylotrophic yeast Pichia pastoris. AB - AIM: Nucleocapsid (N) protein plays an important role in reproduction and pathological reaction of severe acute respiratory syndrome (SARS) coronavirus (SCoV), the antigenicity of the protein is better than spike (S) protein. This study was to find a highly specific and antigenic recombinant SCoV nucleocapsid (rSCoVN) protein, and to provide a basis for further researches on early diagnosis of SARS. METHODS: Full length cDNA of SCoV nucleocapsid (SCoVN) protein was amplified through polymerase chain reaction (PCR) and cloned into yeast expression vector pPIC3.5K to construct plasmid of pPIC3.5K-SCoVN. The plasmid was linearized and then transformed into Pichia pastoris (P.pastoris) GS115 (His Mut+) by electroporation. His(+)Mut(+) recombinant strains were identified by PCR and cultivated on MM/MD plates. The influence of different factors on biomass and rSCoVN protein production during induction phase, such as various induction media, dissolved oxygen (DO) and different final concentrations of methanol, was subsequently studied. The expression level and activation were detected by SDS PAGE and Western-blot respectively. RESULTS: All of the recombinants were His(+)Mut(+) after transformation of P.pastoris with linearized plasmids. The BMMY medium was optimal for recombinant ScoVN (rSCoVN) protein expression and growth of the recombinant strains. The final optimal concentration of methanol was 20 mL/L, the DO had a significant effect on rSCoVN protein expression and growth of recombinant strains. The rSCoVN protein expressed in recombinant strains was about 8% of the total cell protein, 520 mg/L of rSCoVN protein was achieved, and a maximum cell A at 600 nm of 62 was achieved in shake flask culture. The rSCoVN protein had a high specificity against mouse-anti-SARS-CoVN mAb and SARS positive sera, but had no cross-reaction with normal human serum. The biological activity of rSCoVN expressed in P.pastoris was about 4-fold higher than that expressed in E.coli when the same rSCoVN protein quantity was used. CONCLUSION: Active recombinant severe acute respiratory syndrome (SARS) coronavirus nucleocapsid (rSCoVN) protein can be successfully expressed in recombinant methylotrophic yeast P.pastoris GS115. The rSCoVN protein has a high specificity against SARS-CoVN-mAb and SARS positive sera, but has no cross reaction with normal human serum. This provides a basis for further researches on the early diagnosis of SARS and the mechanism of SCoV. PMID- 15534915 TI - Characteristic expression of gamma-aminobutyric acid and glutamate decarboxylase in rat jejunum and its relation to differentiation of epithelial cells. AB - AIM: To investigate the expression between gamma-aminobutyric acid (GABA) and glutamate decarboxylase and its relation with differentiation and maturation of jejunal epithelial cells in rat jejunum. METHODS: Immunohistochemical expression of GABA and glutamate decarboxylase (GAD, including two isoforms, GAD65 and GAD67) was investigated in rat jejunum. Meanwhile, double staining was performed with GAD65 immunohistochemistry, followed by lectin histochemistry of fluorescent wheat germ agglutinin. Furthermore, evaluation of cell kinetics in jejunum was conducted by (3)H-thymidine autoradiography and immunohistochemistry using a monoclonal antibody to proliferating cell nuclear antigen (PCNA). RESULTS: The cells showing positive immunoreactivity GABA and GAD65 were mainly distributed in the villi in rat jejunum, while jejunal epithelial cells were negative for GAD67. Positive GABA or GAD65 staining was mainly located in the cytoplasm and along the brush border of epithelial cells in the middle and upper portions. In addition, a few GABA and GAD65 strongly positive cells were scattered in the upper two thirds of jejunal villi. Double staining showed that GAD65 immunoreactivity was not found in goblet cells. (3)H-thymidine-labeled nuclei were found in the lower and middle portions of jejunal crypts, which was consistent with PCNA staining. Therefore, GABA and GAD65 were expressed in a maturation or functional zone. CONCLUSION: The characteristic expression of GABA and GAD suggests that GABA might be involved in regulation of differentiation and maturation of epithelial cells in rat jejunum. PMID- 15534916 TI - Inhibition of small-intestinal sugar absorption mediated by sodium orthovanadate Na3VO4 in rats and its mechanisms. AB - AIM: To investigate the inhibitory effects of sodium orthovanadate on small intestinal glucose and maltose absorption in rats and its mechanism. METHODS: Normal Wistar rats were lavaged with sodium orthovanadate (16 mg/kg, 4 mg/kg and 1 mg/kg) for 6 d. Blood glucose values were measured after fasting and 0.5, 1, 1.5 and 2 h after glucose and maltose feeding with oxidation-enzyme method. alpha glucosidase was abstracted from the upper small intestine, and its activity was examined. mRNA expression of alpha-glucosidase and glucose-transporter 2 (GLUT2) in epithelial cells of the small intestine was observed by in situ hybridization. RESULTS: Sodium orthovanadate could delay the increase of plasma glucose concentration after glucose and maltose loading, area under curve (AUC) in these groups was lower than that in control group. Sodium orthovanadate at dosages of 10 micromol/L, 100 micromol/L and 1000 micromol/L could suppress the activity of alpha-glucosidase in the small intestine of normal rats, with an inhibition rate of 68.18%, 87.22% and 91.91%, respectively. Sodium orthovanadate reduced mRNA expression of alpha-glucosidase and GLUT2 in epithelial cells of small intestine. CONCLUSION: Sodium orthovanadate can reduce and delay the absorption of glucose and maltose. The mechanism may be that it can inhibit the activity and mRNA expression of alpha-glucosidase, as well as mRNA expression of GLUT2 in small intestine. PMID- 15534917 TI - Gastroprotective effect and mechanism of amtolmetin guacyl in mice. AB - AIM: To investigate the gastroprotective effect and mechanism of amtolmetin guacyl (AMG, MED15) in mice. METHODS: Male and female Kunming strain mice, weighing 18-22 g, were utilized in the experiment. Normal or ethanol-induced gastric mucosal damage models in mice were successfully established to investigate the gastroprotective effect and mechanism of AMG. In the experiment of gastric mucosal damage after repeated treatment with AMG, the mice were randomly divided into 5 groups: normal group, 3 AMG groups receiving (75, 150 and 300 mg/kg), and tolmetin group receiving 90 mg/kg. The mice were randomly divided into 6 groups as follows: normal group, model group, AMG groups with doses of 75, 150 and 300 mg/kg, respectively, and tolmetin group with a dose of 90 mg/kg in ethanol-induced gastric mucosal damage experiment. The severity of gastric mucosal lesions was scored from 0 to 5. Gastric tissue sections were stained with hematoxylin and eosin (HE) and examined under light microscopy. Also gastric tissue sections were stained with uranyl acetate and lead citrate, and examined under electron microscopy. In addition, nitric oxide (NO) and malondialdehyde (MDA) contents, and nitric oxide synthase (NOS) and superoxide dismutase (SOD) activities in the stomach tissue homogenates were measured by biochemical methods. RESULTS: Repeated treatment with AMG (75, 150 and 300 mg/kg) for 7 d did not induce any appreciable mucosal damage, and the average score was not significantly different from that of normal mice. In contrast, tolmetin (90 mg/kg) produced significant gastric mucosal lesions compared with the normal group (P<0.01). AMG (75, 150 and 300 mg/kg) significantly reduced the severity of gastric lesions induced by ethanol in a dose-dependent manner as compared with the model group (P<0.05, AMG 75 and 150 mg/kg vs model; P<0.01, AMG 300 mg/kg vs model). Light and electron microscopy revealed that AMG (150 and 300 mg/kg) induced minimal changes in the surface epithelium layer, without vascular congestion or leucocyte adherence. AMG (75,150 and 300 mg/kg) demonstrated dose dependent gastroprotective effects on mice in our study. AMG (75, 150 and 300 mg/kg) could significantly increase NO content and NOS level in the stomach homogenates of mice compared with the model group (P<0.05, AMG 75 mg/kg and 150 mg/kg groups vs model group; P<0.01, AMG 300 mg/kg vs model group) respectively. Moreover, AMG (150 and 300 mg/kg) not only significantly increased SOD activities but also obviously decreased the MDA content in the stomach homogenates of mice. PMID- 15534918 TI - Dynamic changes in the expression of matrix metalloproteinases and their inhibitors, TIMPs, during hepatic fibrosis induced by alcohol in rats. AB - AIM: To determine the dynamic changes in the expression of matrix metalloproteinases (MMPs) and the endogenous tissue inhibitors of MMPs inhibitors (TIMPs) during hepatic fibrosis induced by alcohol. METHODS: Male Sprague-Dawley rats were randomly divided into normal, 4 d, 2 wk, 4 wk, 9 wk and 11 wk groups, and the model rats were fed with a mixture of alcohol by gastric infusion at the designed time, respectively, then decollated and their livers were harvested for the examination of MMP-2, MMP-3, MMP-9, MMP-13, TIMP-1 and TIMP-2 by immunohistochemistry, zymograghy and Western blotting, respectively. RESULTS: Normal rats had moderate expression of MMP-2, which was decreased in the model rats except in the 11 wk group, where MMP-2 expression slightly increased. MMP-3 had the similar changing pattern to MMP-2 despite weaker expression. MMP-9 expression decreased in the 4 d and 2 wk groups, rose in the 4 wk group, decreased again in the 9 wk group and returned to normal levels in the 11 wk group. MMP-13 expression decreased in the 4 d and 2 wk groups, and returned to normal levels in the 4 wk, 9 wk and 11 wk groups. TIMP-1 expression decreased in the 4 d and 2 wk groups, but sharply increased in the 4 wk group and sustained at a high level even after modeling was stopped for 2 wk. In normal rats TIMP-2 expression was strong. However, it decreased as soon as modeling began, and then gradually rose, but remained to a level lower than that in normal rats even after modeling was stopped for 2 wk. CONCLUSION: MMP-2 may not always expresses at a high level during hepatic fibrosis. MMP-13 and MMP-3 are acutely affected by TIMP 1. In this model TIMP-1 is the most powerful factor imposed on capillarization and peri-sinusoidal fibrosis. TIMP-2 is the most effective regulator on the metabolism of type IV collagen located in the basement of sinus. PMID- 15534919 TI - Scintigraphic detection of carcinoid tumors with a cost effectiveness analysis. AB - AIM: To evaluate the diagnostic sensitivity and accuracy and the cost effectiveness of this technique in the detection of gastroenteropancreatic carcinoid tumors and their metastases in comparison with conventional imaging methods. METHODS: Somatostatin receptor scintigraphy (SRS) was performed in 24 patients with confirmed carcinoids and 7 under investigation. The results were compared with those of conventional imaging methods (chest X-ray, upper abdominal ultrasound, chest CT, upper and lower abdominal CT). Also a cost-effectiveness analysis was performed comparing the cost in Euro of several combinations of SRS with conventional imaging modalities. RESULTS: SRS visualized primary or metastatic sites in 71.0% of cases and 61.3% of conventional imagings. The diagnostic sensitivity of the method was higher in patients with suspected lesions (85.7% vs 57.1%). SRS was less sensitive in the detection of metastatic sites (78.9% vs 84.2%). The undetectable lesions by SRS metastatic sites were all in the liver. Between several imaging combinations, the combinations of chest X ray/upper abdominal CT/SRS and chest CT/upper abdominal CT/SRS showed the highest sensitivity (88.75%) in terms of the number of detected lesions. The combinations of chest X-ray/upper abdominal US/SRS and chest CT/upper abdominal ultrasound /SRS yielded also a quite similar sensitivity (82%). Compared to the cost of the four sensitive combinations the combination of chest X-ray/upper abdominal ultrasound/SRS presented the lower cost, 1183.99 Euro vs 1251.75 Euro for chest CT/upper abdominal ultrasound/SRS, 1294.93 Euro for chest X/ray/upper abdominal CT/SRS and 1362.75 Euro for chest CT/upper abdominal CT/SRS. CONCLUSION: SRS imaging is a very sensitive method for the detection of gastroenteropancreatic carcinoids but is less sensitive than ultrasound and CT in the detection of liver metastases. Between several imaging combinations, the combination of chest X ray/upper abdominal CT/SRS shows the highest sensitivity with a cost of 1294.93 Euro. PMID- 15534921 TI - Monocyte chemotactic protein-1 and soluble adhesion molecules as possible prognostic markers of the efficacy of antiviral treatment in chronic hepatitis C. AB - AIM: To explain the role of Monocyte chemotactic protein-1 (MCP-1) and soluble adhesion molecules in chronic hepatitis C during the treatment of interferon alpha (IFNalpha) 2 b and ribavirin (RBV). METHODS: Concentrations of MCP-1, soluble adhesion molecules intercellular adhesion molecule-1 (sICAM-1), sP selectin, interleukin (IL) 6, and IL10 in serum were estimated in the group of 40 patients with chronic hepatitis C treated with IFNalpha2 b and RBV in 0, 16, 32, 48 wk of the therapy. RESULTS: In chronic hepatitis C, before and during the treatment, the serum levels of MCP-1 and sP-selectin in responders were similar to those of healthy subjects. In non-responders (NR), MCP-1 increased in the course of IFNalpha+RBV treatment, differences were statistically significant as compared to responders. MCP-1 correlated statistically with the activity of periportal inflammation (r = 0.35, P<0.05) but not with staging of liver fibrosis. sICAM-1 positively correlated with inflammatory activity and fibrosis in NR. sP-selectin did not correlate with histological findings in the liver. The MCP-1 correlated with the soluble form of sP-selectin concentrations (r = 6, P<0.001) and with IL-10 level in NR (r = 0.4, P<0.05). There was no correlation observed between the concentration of MCP-1 and sICAM-1, IL-6 during the treatment. CONCLUSION: MCP-1 concentration may be a prognostic marker of the efficacy of IFN+RBV therapy in patients with chronic hepatitis C. PMID- 15534920 TI - Intra-tumor injection of H101, a recombinant adenovirus, in combination with chemotherapy in patients with advanced cancers: a pilot phase II clinical trial. AB - AIM: H101, an E1B 55 kD gene deleted adenovirus, has been shown to possess oncolysis activity experimentally and proved to be safe in preliminary phase I study. The current study was designed to evaluate its anti-tumor activity and toxicity in combination with chemotherapy in patients with late stage cancers. METHODS: H101 5.0x10(11) virus particles were given by intra-tumor injection daily for five consecutive days at every three-week cycle, combined with routine chemotherapy, to one of the tumor lesions of 50 patients with different malignant tumors. Tumor lesions without H101 injection in the same individuals were used as controls. The efficacy and toxicity were recorded. RESULTS: Forty-six patients were evaluable with a 30.4% response rate. H101 injection in combination with chemotherapy induced three complete response (CR) and 11 partial response (PR), giving an overall response rate of 28.0% (14/50) among intention-to-treat patients. The response rate for the control lesions was 13.0%, including one case with CR and five cases with PR, which was significantly lower than that for the injected lesions (P<0.05). Main side effects were fever (30.2%) and pain at the injected sites (26.9%). Grade 1 hepatic dysfunction was found in four patients, grade 2 in one patient, and grade 4 in one patient. Hematological toxicity (grade 4) was found in four patients. CONCLUSION: Intra-tumor injection of the genetically engineered adenovirus H101 exhibits potential anti-tumor activity to refractory malignant tumors in combination with chemotherapy. Low toxicity and good tolerance of patients to H101were observed. PMID- 15534922 TI - Interleukin (IL) 5 levels and eosinophilia in patients with intestinal parasitic diseases. AB - AIM: Intestinal parasitic diseases are commonly accompanied with diarrhoeal symptoms and allergic reactions. Eosinophilia occurs as a result of IL-5 synthesized from Th2 cells during allergic reactions. IL-5 acts as a factor activating eosinophils. The aim of this study was to compare the IL-5 cytokine measurements in serum samples and cell cultures. And also to compare eosinophilia observed in helminth infections and protozoon infections accompanied with allergy. METHODS: Twenty-three patients who presented with diarrhoeal symptoms and allergic complaints were tested positive for intestinal parasites, as well as 21 controls with allergic complaints who did not have any intestinal parasites were included in this study. IL-5 production in in vitro cell cultures prepared by using phytohemaglutinin (PHA) to stimulate peripheral blood mononuclear cells (PBMC) obtained from the blood samples taken from these patients were compared with the IL-5 level in serum. Furthermore, the IL-5 production in protozoon and helminth infections was also compared. Absolute eosinophil values in 1 mm(3) of blood were calculated by means of peripheral smear in both groups within the scope of the study. RESULTS: Parasites such as helminth detected in 15 (65.2%) and protozoon in 8 (34.8%) of the patients were included in this study. As regards the values of the sera in both patients with parasite infection and controls, the IL-5 production was found to be higher in the cell culture supernatant (P<0.001 and P<0.05). When the IL-5 level of the patients with helminth parasites was compared with that of those with protozoon, it was determined that the IL-5 level in serum was more significant in the patients with protozoon than in those with helminth (P<0.05). In the study group, the patients were found to have parasites, the percentage of eosinophil was 7.0% compared to 6.5% in the control group. Thus, there was no significant difference between the eosinophil values (P>0.05). CONCLUSION: It was found that IL-5 cytokine levels in serum samples from the patients with helminth and protozoon displayed more measurable values as compared to the IL-5 levels after stimulation with mitogen. It is concluded that IL-5 acts as a triggering factor in the toxiallergic complaints commonly seen in helminth and protozoon infections. PMID- 15534923 TI - Diagnosis of abdominal tuberculosis: experience from 11 cases and review of the literature. AB - AIM: To analyze the experience within our hospital and to review the literature so as to establish the best means of diagnosis of abdominal tuberculosis. METHODS: The records of 11 patients (4 males, 7 females, mean age 39 years, range 18-65 years) diagnosed with abdominal tuberculosis in Harran University Hospital between January 1996 and October 2003 were analyzed retrospectively and the literature was reviewed. RESULTS: Ascites was present in all cases. Other common findings were weight loss (81%), weakness (81%), abdominal mass (72%), abdominal pain (72%), abdominal distension (63%), anorexia (45%) and night sweat (36%). The average hemoglobin was 8.2 g/dL and the average ESR was 50 mm/h (range 30-125). Elevated levels of cancer antigen CA-125 were determined in four patients. Abdominal ultrasound showed abnormalities in all cases: ascites in all, tuboovarian mass in five, omental thickening in 3, and enlarged lymph nodes (mesenteric, para-aortic) in 2. CT scans showed ascites in all, pelvic mass in 5, retroperitoneal lymphadenopathy in 4, mesenteric stranding in 4, omental stranding in 3, bowel wall thickening in 2 and mesenteric lymphadenopathy in 2. Only one patient had a chest radiograph suggestive of a new TB lesion. Two had a positive family history of pulmonary TB. None had acid-fast bacilli (AFB) in the sputum and the tuberculin test was positive in only two. Laparotomy was performed in 6 cases, laparoscopy in 4 and ultrasound-guided fine needle aspiration in 2. In those patients subjected to operation, the findings were multiple diffuse involvement of the visceral and parietal peritoneum, white 'miliary nodules' or plaques, enlarged lymph nodes, ascites, 'violin string' fibrinous strands, and omental thickening. Biopsy specimens showed granulomas, while ascitic fluid showed numerous lymphocytes. Both were negative for acid-fast bacilli by staining. PCR of ascitic fluid was positive for Mycobacterium tuberculosis (M. tuberculosis) in all cases. CONCLUSION: Abdominal TB should be considered in all cases with ascites. Our experience suggests that PCR of ascitic fluid obtained by ultrasound-guided fine needle aspiration is a reliable method for its diagnosis and should at least be attempted before surgical intervention. PMID- 15534924 TI - Serum concentration of sFas and sFasL in healthy HBsAg carriers, chronic viral hepatitis B and C patients. AB - AIM: To estimate the amount of apoptosis among healthy HBsAg carriers, patients with chronic HBV infection treated with lamivudine and patients with chronic HCV infection treated with interferon alpha and ribavirin. Activity of apoptosis was evaluated by serum sFas/sFasL concentration measurement. Moreover dependence between apoptosis and HBV-DNA or HCV-RNA levels was studied. METHODS: Eighty-six persons were included into study: 34 healthy HBsAg carriers, 33 patients with chronic HBV infection and 19 patients with chronic HCV infection. Serum levels of sFas/sFasL were measured by ELISA assay. HBV-DNA and HCV-RNA were measured by RT PCR assay. Levels of sFas/sFasL were determined before and 2 and 12 wk after therapy in patients with chronic hepatitis B and C infection. HBV-DNA or HCV-RNA was detected before treatment and 6 mo after treatment. RESULTS: Twenty-four (71%) healthy HBsAg carriers showed HBV-DNA over 10(5)/mL, which was comparable to the patients with chronic hepatitis B. Independently from HBV-DNA levels, the concentration of sFas among healthy HBsAg carriers was comparable to healthy persons. Among patients with chronic hepatitis B and C, the concentration of sFas was significantly higher in comparison to healthy HBsAg carriers and healthy persons. In chronic hepatitis B patients the concentration of sFas was decreased during lamivudine treatment. Among chronic hepatitis C patients the concentration of sFas was increased during IFN alpha and ribavirin treatment. sFasL was not detected in control group. Furthermore sFasL occurred more frequently in chronic hepatitis C patients in comparison to chronic hepatitis B patients. CONCLUSION: There are no correlations between apoptosis and HBV-DNA levels. However ther is an association between apoptosis and activity of inflammation in patients with chronic HBV infection. Apoptosis can be increased in patients with chronic hepatitis C by effective treatment which may be a result of apoptosis stimulation by IFN-alpha. PMID- 15534925 TI - In vivo suppressive effect of nuclear factor-kappaB inhibitor on neutrophilic inflammation of grafts after orthotopic liver transplantation in rats. AB - AIM: To investigate the effect of pyrrolidine dithiocarbamate (PDTC), a novel nuclear factor-kappaB (NF-kappaB) inhibitor, on expression of multiple inflammatory mediators and neutrophilic inflammation of cold preserved grafts after rat liver transplantation and its significance. METHODS: Orthotopic liver transplantation (OLT) was performed after 24 h of cold storage using University of Wisconsin solution with varied concentrations of PDTC. We determined the time course of NF-kappaB activation and expression of multiple inflammatory signals, such as tumor necrosis factor-alpha (TNF-alpha), cytokine-inducible neutrophil chemoattractant (CINC), and intercellular adhesion molecule-1 (ICAM-1) by ELISA methods. Serum alanine aminotransferase (ALT), intrahepatic myeloperoxidase (MPO)/WBC (a measure of neutrophil accumulation) and Mac-1 expression (a measure of circulating neutrophil activity) were also evaluated. RESULTS: PDTC decreased NF-kappaB activation induced by prolonged cold preservation in a dose dependent manner (from 20 mmol/L to 60 mmol/L), diminished TNF-alpha, CINC, ICAM-1 proteins in the grafts, and reduced the expression of increases in plasma TNF-alpha levels induced by prolonged cold preservation. Neutrophilic inflammation of the graft was significantly suppressed after preservation with PDTC (P<0.05). The total neutrophil accumulation in PDTC (40 mmol/L) group (7.04+/-0.97) was markedly reduced compared to control group (14.07+/-1.31) (P<0.05). Mac-1 expression was significantly reduced in PDTC (40 mmol/L) group (181+/-11.3%) compared with the control group (281+/-13.2%) (P<0.05) at 6 h after reperfusion. Furthermore, PDTC inhibited the increased serum ALT levels after liver transplantation. CONCLUSION: PDTC can inhibit B NF-kappaB activation and expression of the inflammatory mediators, which are associated with improved graft viability via inhibiting intrahepatic neutrophilic inflammation. Our study suggests that a therapeutic strategy directed at inhibition of NF-kappaB activation in the transplanted liver might be effective in reducing intrahepatic neutrophilic inflammation, and would be beneficial to cold preserved grafts. PMID- 15534926 TI - Clinical analysis of eight kindreds of familial adenomatous polyposis. AB - AIM: To study the early diagnosis and management of familial adenomatous polyposis (FAP). METHODS: Eight pedigrees of FAP were collected and their pedigree trees were protracted. Clinical characteristics and treatment outcomes of FAP patients in these kindreds were analysed. RESULTS: A total of 157 members were investigated in eight kindreds and 25 patients with FAP were diagnosed. The ratio of male patients and female patients was 16:9 and the average age at onset was 38 years. Among them, six patients died of cancer with a mortality rate of 28%, and 36% (9/25) FAP patients were diagnosed as synchronous colorectal cancer on the basis of FAP. A proband was diagnosed as synchronous colorectal cancer with liver metastasis and died 11 mo later after partial colectomy and hepatic metastatic lesion biopsy. The other seven probands received total abdominal colectomy and rectal mucosectomy with ileal pouch-anal anastomosis (IPAA), and one of them was diagnosed as synchronous colon cancer on the basis of FAP and was still alive after 7.5 years follow-up. Among the other seven patients with synchronous colorectal cancer on the basis of FAP underwent total abdominal colectomy with ileorectal anastomosis (IRA), one underwent total remnant rectum resection and ileostomy for recurrent carcinoma in the retained rectum 2.5 years later after the IRA and was still alive, while the others all died of recurrence with a median survival time of 4.6 years. Through close follow-up and termly endoscopic surveillance, three FAP patients were detected before presenting symptoms at the age of 18, 20 and 23 years, respectively. Prophylactic IPAA was performed and results were satisfactory after the patients were followed-up for 6, 1, and 8 years, respectively. CONCLUSION: Pedigree investigation, close follow up and termly endoscopic surveillance are very important for early detection of FAP. Prophylactic IPAA can give satisfactory results to FAP patients. PMID- 15534927 TI - Effects of c-myb antisense RNA on TGF-beta1 and beta1-I collagen expression in cultured hepatic stellate cells. AB - AIM: To investigate the effects of c-myb antisense RNA on cell proliferation and the expression of c-myb, TGF-beta1 and beta1-I collagen in cultured hepatic stellate cells (HSC) from rats. METHODS: Recombinant retroviral vector of c-myb antisense gene (pDOR-myb) was constructed, and then transfected into retroviral package cell line PA317 by means of DOTAP. The pseudoviruses produced from the resistant PA317 cells were selected with G418 to infect HSCs isolated from rat livers. The cell proliferation was measured by 3-[4, 5-Dimethylthiazolzyl]-2, 5 diphenyl tetrazo-dium bromide(MTT) method. The expression of c-myb, alpha (1)-I collagen and TGF-beta1 mRNA, and c-myb protein in HSCs was detected with semi quantitive reverse transeription-polymerase chain reaction (RT-PCR) and Western blot respectively. RESULTS: HSCs from rats were isolated successfully with the viability >98%. In the pDOR-myb infected HSCs, the c-myb protein expression, cell proliferation,and alpha (1)-I collagen and TGF-beta1 mRNA expression were repressed significantly compared with their corresponding control groups (P<0.01). CONCLUSION: c-myb plays a key role in activation and proliferation of HSC. c-myb antisense RNA can inhibit cell proliferation, alpha (1)-I collagen and TGF-beta1 mRNA expression, suggesting that inhibition of c-myb gene expression might be a potential way for the treatment of liver fibrosis. PMID- 15534928 TI - Intravenous pantoprazole versus ranitidine for prevention of rebleeding after endoscopic hemostasis of bleeding peptic ulcers. AB - AIM: The role of intravenous pantoprazole in treatment of patients with high-risk bleeding peptic ulcers following endoscopic hemostasis remains uncertain. We therefore conducted the pilot prospective randomized study to assess whether intravenous pantoprazole could improve the efficacy of H(2)-antagonist as an adjunct treatment following endoscopic injection therapy for bleeding ulcers. METHODS: Patients with active bleeding ulcers or ulcers with major signs of recent bleeding were treated with distilled water injection. After hemostasis was achieved, they were randomly assigned to receive intravenous pantoprazole or ranitidine. RESULTS: One hundred and two patients were enrolled in this prospective trial. Bleeding recurred in 2 patients (4%) in the pantoprazole group (n = 52), as compared with 8 (16%) in the ranitidine group (n = 50). The rebleeding rate was significantly lower in the pantoprazole group (P = 0.04). There were no statistically significant differences between the groups with regard to the need for emergency surgery (0% vs 2%), transfusion requirements (4.9+/-5.9 vs 5.7+/-6.8 units), hospital days (5.9+/-3.2 vs 7.5+/-5.0 d) or mortality (2% vs 2%). CONCLUSION: Pantoprozole is superior to ranitidine as an adjunct treatment to endoscopic injection therapy in high-risk bleeding ulcers. PMID- 15534929 TI - Inhibition of transfected PTEN on human colon cancer. AB - AIM: To study the inhibitory effect of transfected PTEN on LoVo cells. METHODS: Human PTEN cDNA was transferred into LoVo cells via lipofectin and PTEN mRNA levels and its expression were analyzed by Western blot and flow cytometry. Before or after transfection, the effects of 5-Fu on inhibiting cell proliferation and inducing apoptosis were measured by flow cytometry, DNA bands and MTT. RESULTS: PTEN transfection significantly up-regulated PTEN expression in LoVo cells. 5-Fu inhibited cell proliferation and induced apoptosis in transfected LoVo cells. CONCLUSION: Transfected PTEN can remarkably up-regulate PTEN expression in LoVo cells and promote the apoptosis. PTEN transfection is associated with 5-Fu treatment effect and has a cooperatively cytotoxic effect. PMID- 15534930 TI - Serum sIL-2R, TNF-alpha and IFN-gamma in alveolar echinococcosis. AB - AIM: to approach the relationship between alveolar echinococcosis (AE) pathology and level of sIL-2R,TNF-alpha and IFN-gamma in sera and the significance of cytokines in development of AE. METHODS: After 23 patients with AE were confirmed by ELISA and ultrasound, their sera were collected and the concentrations of sIL 2R,TNF-alpha and IFN-gamma were detected by double antibody sandwich. Twelve healthy adults served as controls. According to the status of livers of AE patients by ultrasound scanning, they were divided into 4 groups: P(2), P(3), P(4) groups and C group (control). Average of concentrations of sIL-2R,TNF-alpha and IFN-gamma in homologous group was statistically analyzed by both ANOV and Newman-Keuls, respectively. RESULTS: The mean of sIL-2R in P(2) group was 97+/ 29, P(3): 226+/-80, P(4): 194+/-23 and control group (111+/-30)X10(3) u/L (P<0.01). The mean of TNF-alpha in P(2) group was 1.12+/-0.20, P(3): 3.67+/-1.96, P(4): 1.30+/-0.25 and control group 0.40+/-0.19 mug/L (P<0.01). The mean of IFN gamma in P(2) group was 360+/-20, P(3): 486+/-15, P(4): 259+/-19 and control group: 16+/-2 ng/L (P<0.01). Judged by ANOV and Newman-Keuls, the mean concentrations of sIL-2R, TNF-alpha and IFN-gamma had a significant difference among groups. Except for P(2) group, the mean sIL-2R between other groups of AE patients had a significant difference (P<0.05). The mean of TNF-alpha concentration in P(3) group was the highest (P<0.01). The mean of IFN-gamma concentration in all patients was higher than that in control group (P<0.01), but there was no difference between AE groups (P>0.05). CONCLUSION: Low sIL-2R level indicates an early stage of AE or stable status, per contra, a progression stage. Higher level of TNF-alpha might be related to the lesion of liver. The role of single IFN-gamma is limited in immunological defense against AE and it can not fully block pathological progression. PMID- 15534931 TI - Effect of arsenic trioxide on human hepatocarcinoma in nude mice. AB - AIM: To study the effect of arsenic trioxide (As(2)0(3)) on human hepatoma cell line BEL-7402 in vivo. METHODS: Human hepatoma cell line BEL-7402 cultured in vitro was inoculated into nude mice and arsenic trioxide, 5-Fu and saline were injected into abdominal cavity of the nude mice respectively. The volumes of tumor and general conditions of the nude mice and structural changes of the liver and kidney were observed. Morphologic changes were studied under electron microscope. Expression of AFP was investigated by immunohistochemical method. RESULTS: As(2)O(3) could inhibit the growth of tumor. The tumor growth inhibitory rate in mice treated with 2.5 mg/kg As(2)O(3) was 53.42% on the tenth day. The tumor growth inhibitory rate in mice treated with 5 mg/kg As(2)O(3) was 79.28% on the fifth day and 96.58% on the tenth day respectively. As(2)O(3) did not damage the liver and kidney of nude mice, or affect the blood system. Typical apoptotic morphological changes were found under electron microscope, and the change of mitochondria was obvious. The expression rate of AFP declined after treatment. CONCLUSION: Arsenic trioxide can induce apoptosis of human hepatoma cells, and inhibit proliferation of tumor with no obvious side effects on liver and kidney. PMID- 15534932 TI - Primary small cell carcinoma of esophagus: report of 9 cases and review of literature. AB - AIM: To analyze the clinical manifestations, pathological features and treatment of primary small cell carcinoma (SCC) of the esophagus and to review the literature on this entity. METHODS: The records of 9 patients with primary esophageal small cell carcinoma were examined and the demographic data, presenting symptoms, methods of tumor diagnosis, and types of treatment given, response to treatment, pathologic findings, and clinical outcome were reviewed. Features of mixed patterns of histological differentiation and lymph node metastases were specifically sought. RESULTS: All the patients reported dysphagia, weight loss and chest pain as the initial symptoms. In 5 cases the tumors were located in the mid-esophagus, 3 cases in the lower third of the esophagus and 1 case in the upper third. The average length of esophageal involvement was 5 cm. They underwent radical resection, regional lymph node clearance and esophageal-stomach anastomosis in thorax or at neck. Two patients had a stage IIa disease, five had a stage IIb disease, and the other two had a stage III disease of International Union Contrele Cancer (UICC). All of them were histologically and immunohistochemically confirmed SCC of esophagus. Immunohistochemical staining for neuron-specific enolase (NSE), synaptophsin (Syn) and chromogranin A exhibited strong immunoreactivity in all specimens. Three of the nine resected specimens showed foci of squamous cell carcinoma in situ. Metastasis was present in 7 of 9 adjacent lymph nodes. All the patients survived the operations and made an uneventful postoperative recovery. They received adjuvant systemic chemotherapy and local radiation therapy after discharge. During follow-up, three patients developed multiple liver, brain, lung and bone metastases and died between 5 and 18 mo after the diagnosis. Three patients developed widespread metastasis disease and died between 18 and 37 mo after the diagnosis. There was no local tumor recurrence in these 6 patients. The other three patients were lost during follow-up. CONCLUSION: Primary small cell carcinoma of the esophagus is a rare but very malignant tumor. Radical resection combined with chemotherapy and radiotherapy is helpful in limited stage cases. PMID- 15534933 TI - Integration of E. coli aroG-pheA tandem genes into Corynebacterium glutamicum tyrA locus and its effect on L-phenylalanine biosynthesis. AB - AIM: To study the effect of integration of tandem aroG-pheA genes into the tyrA locus of Corynebacterium glutamicum (C. glutamicum) on the production of L phenylalanine. METHODS: By nitrosoguanidine mutagenesis, five p fluorophenylalanine (FP)-resistant mutants of C.glutamicum FP were selected. The tyrA gene encoding prephenate dehydrogenase (PDH) of C.glutamicum was amplified by polymerase chain reaction (PCR) and cloned on the plasmid pPR. Kanamycin resistance gene (Km) and the P(BF) -aroG-pheA-T (GA) fragment of pGA were inserted into tyrA gene to form targeting vectors pTK and pTGAK, respectively. Then, they were transformed into C.glutamicum FP respectively by electroporation. Cultures were screened by a medium containing kanamycin and detected by PCR and phenotype analysis. The transformed strains were used for L-phenylalanine fermentation and enzyme assays. RESULTS: Engineering strains of C.glutamicum (Tyr(-)) were obtained. Compared with the original strain, the transformed strain C. glutamicum GAK was observed to have the highest elevation of L-phenylalanine production by a 1.71-fold, and 2.9-, 3.36-, and 3.0-fold in enzyme activities of chorismate mutase, prephenate dehydratase and 3-deoxy-D-arabinoheptulosonate-7 phosphate synthase, respectively. CONCLUSION: Integration of tandem aroG-pheA genes into tyrA locus of C. glutamicum chromosome can disrupt tyrA gene and increase the yield of L-phenylalanine production. PMID- 15534934 TI - Co-mutation of p53, K-ras genes and accumulation of p53 protein and its correlation to clinicopathological features in rectal cancer. AB - AIM: To determine the accuracy of p53 gene mutations predicted by overexpression of p53 protein immunohistochemically, and to investigate the co-mutation of p53 and K-ras genes in rectal cancer and its effect on promoting malignant biologic behaviors of tumors. METHODS: Ninety-seven specimens of rectal cancer were surgically resected in our hospital from August 1996 to October 1997. The hot mutation areas of p53 gene (in exons 5-8) and K-ras gene (in codon 5/12 and 13) were detected with polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), and overexpression of p53 protein was detected with immunohistochemistry (IHC) in the 97 specimens of rectal cancer. Correlation between gene mutations and tumor clinicopathologic factors was studied, and survival analysis was penfomed as well. RESULTS: There were 36 cases of p53 gene mutations in 61 p53 protein positive cases, and 21 cases of p53 gene non-mutation in 36 p53 protein negative cases respectively. The coincidence rate of p53 gene mutation by IHC method with PCR-SSCP method was 58.8% (57/97). The mutation rate of p53 gene was 52.6% (51/97), while K-ras gene mutation was observed in codons 12 and 13 in 61 cases with a mutation rate of 62.9% (61/97). Single gene mutation of p53 or K-ras was found in 32 cases. Both p53 and K-ras gene mutation were found in 48 cases. Statistical analysis showed that p53 and K-ras gene mutations were not related to the clinicopathologic factors, including tumor size, gross tumor type, histological classification, differentiation, invasion to intestinal veins, lymphatics and nerves, invasive depth to wall, lymph node metastasis, and Dukes' stages (P>0.05). The survival in patients with no gene mutation, single gene mutation and both gene mutations were similar (P>0.05). CONCLUSION: IHC has a certain false positive and false negative rate in detecting p53 gene mutations. Malignant biological behaviours of rectal cancer are not enhanced by p53 and K ras gene mutations. Co-mutation of p53 and K-ras gene has neither synergic carcinogenesis-promoting effect, nor prognostic effect on rectal cancer. PMID- 15534935 TI - Diagnostic evaluation of acute pancreatitis in two patients with hypertriglyceridemia. AB - We present two diagnostically challenging cases of acute pancreatitis with hypertriglyceridemia accompanied with chylomicronemia caused with a deficiency of lipoprotein lipase and with the presence of type V hyperlipidemia. Both cases suffered from acute abdomen following the ingestion of fatty food and revealed the increase in parameters of inflammation without significant elevation of serum amylase levels. The imaging examination of ultrasonography could not detect significant findings of acute pancreatitis and a computer tomography scan eventually confirmed the findings of acute pancreatitis. Both cases responded to a low fat diet and administration of a cholecystokinin receptor antagonist, exhibiting a relief of abdominal symptoms. As in the present cases with acute abdomen following the ingestion of fatty food, the identification of serum hypertriglyceridemia and an abdominal computer tomography scan might be useful in establishing the diagnosis of acute pancreatitis and in developing the therapeutic regimen, when hypertriglyceridemia interferes with the evaluation of pancreatic enzyme activities and ultrasound examination provides poor pancreatic visualization. PMID- 15534936 TI - Antiretroviral rollout in South Africa fails to meet demands. PMID- 15534937 TI - WHO's tuberculosis control strategy said to be insufficient. PMID- 15534938 TI - MMR debate fall-out continues. PMID- 15534939 TI - USA to manufacture two million doses of pandemic flu vaccine. PMID- 15534940 TI - Melioidosis strikes Singapore. PMID- 15534941 TI - Nanowires show potential as virus detectors. PMID- 15534943 TI - New vaccines to fight killer rotavirus. PMID- 15534944 TI - Disgruntled doctors. PMID- 15534945 TI - Dealing in drugs. PMID- 15534946 TI - [Implication of genes of the serotonergic system on vulnerability to suicidal behavior]. AB - There are many risk factors associated with vulnerability to suicidal behaviour, and the results of family studies, twin studies and adoption studies suggest that they include a genetic predisposition. Moreover, this gentic susceptibility may be specific and independent of the genetic susceptibility to psychiatric disorders associated with suicidal behaviour (e.g., bipolar disorders, schizophrenia, alcoholism). Several groups have carried out association studies using a "candidate gene strategy", with the goal of identifying the genes involved in susceptibility to suicidal behavior. There is compelling evidence from research in biological psychiatry that abnormalities in the functioning of the central serotonergic system are involved in the pathogenesis of suicidal behavior, and the results of association studies suggest that the gene coding for tryptophan hydroxylase, which is the serotonin synthesis enzyme, and the serotonin transporter gene are involved in susceptibility to suicidal behavior. Furthermore, these genes may influence the suicidal phenotype through different gene-gene interactions and gene-early environment interactions. Current studies aim to identify either the precise phenotypes associated with genes for vulnerability to suicidal behaviour or the intermediate phenotypes (e.g., impulsivity, anger dyscontrol) associated with these genes. PMID- 15534947 TI - [Interdisciplinarity in reconstructive surgery of the extremities]. AB - From the perspective of trauma and orthopedic surgery interdisciplinary reconstructive surgery of the extremities encompasses different indications ranging from IIIB/Copen fractures with major segmental loss of bone and soft tissue, to arterial vessel in-jury necessitating vascular repair and to biological, plastic reconstructions following resections of musculoskeletal tumors. The interdisciplinary treatment concept including trauma-orthopedic surgery combined with vascular, plastic and neuro- as well as microsurgery has significantly decreased amputation rates and functional deficits thereby improving quality of life and long-term oncological outcome The multisdisciplinary management of both complex trauma and malignant bone/ soft tissue sarcoma of the extremity is an integral task of the surgical trauma or tumor center. Close interdisciplinary communication and expierence are the precondition for performance of a priority-adapted therapeutic strategy, low complication rates and improvement of overall prognosis. PMID- 15534949 TI - Advances in colon cancer surgery reported. PMID- 15534948 TI - Enlarged prostate. Know when to seek treatment. PMID- 15534951 TI - Secondary hypertension. High blood pressure with a specific cause. PMID- 15534950 TI - Bone health and phosphorus intake studied. PMID- 15534952 TI - Lupus. Usually manageable. PMID- 15534953 TI - Dengue fever. World travelers beware. PMID- 15534954 TI - I stopped taking hormone replacement therapy a year and a half ago after reports that there might be health risks associated with it. Now that I've been off hormones, do my health risks improve? PMID- 15534955 TI - I've been battling frequent constipation and have been taking a stool softener a few times a week. Is it safe to use them this often? PMID- 15534956 TI - Publishing and getting published. PMID- 15534957 TI - Utilising clinical expertise in postgraduate nursing education. PMID- 15534958 TI - Keeping track: electronic health records. PMID- 15534959 TI - Biological monitoring of exposure to toluene diisocyanate. AB - OBJECTIVES: Toluene diisocyanate (TDI) is used in the manufacture of polyurethane and is a potent inducer of diseases of the airways. In this study, 2,4- and 2,6 toluenediamine in hydrolyzed urine and plasma were evaluated as biomarkers of exposure to 2,4- and 2,6-TDI, respectively. METHODS: For 81 exposed workers from nine different plants, the personal 8-hour time-weighted-average exposure to TDI was monitored by a filter method with 1-(2-methoxyphenyl)piperazine. In parallel, urinary samples (U1) were collected during the last 4 hours of the workshift. On a different occasion, blood samples and additional urinary samples (U2) were collected from the exposed workers, and also from a reference group consisting of 121 unexposed workers. The biomarker levels were determined in urine and plasma by the use of alkaline hydrolysis. RESULTS: There were strong associations between the personal air and biomarker levels, with correlation coefficients in the range of 0.75-0.88 for the U1 samples and in the range of 0.50-0.78 for the plasma samples. By weighted linear regression, the relations were calculated between the air and biomarker levels. The slopes of the obtained regression curves ranged from 1.8 to 2.7 m3/1 for air-urine and from 2.2 to 2.9 m3/1 for air plasma, and the intercepts were all close to the origin of the coordinates. Through the extrapolation of these regression curves, biological exposure limits were calculated. CONCLUSIONS: The biological monitoring methods and strategies presented in this report are useful for assessing exposure to TDI in practice. PMID- 15534960 TI - [Patterns of clinical course in persons with mental disorders: which impact on social disability and quality of life? A longitudinal study]. AB - AIMS: To investigate in persons with mental disorders 1) the patterns of clinical course and their frequencies, 2) the impact of clinical course on two social dimensions of outcome, such as disability and quality of life. METHODS: Study conducted with a longitudinal design in the "real world" of community mental health services. Clinical course was retrospectively assessed by using an instrument developed by our group, taking into account previous literature in this area; disability and quality of life were measured, respectively, with the WHO-Disability Assessment Schedule and the Lancashire Quality of Life Profile. RESULTS: In patients with non affective psychosis, continuous and episodic course showed the same tendency to occur, whereas in subjects with affective disorders (either psychotic or neurotic) episodic course was more frequent. Continuous course was associated with higher levels of disability and lower quality of life in psychotic patients, while a poorer quality of life in some areas was associated with episodic course in patients with non psychotic disorders. CONCLUSIONS: The impact of clinical course on social disability and quality of life is different depending upon the specific diagnostic category. This suggests that specific and individualised interventions should be provided in order to prevent the negative impact of clinical course on life conditions of persons with mental disorders. PMID- 15534961 TI - Necrotizing soft tissue infections: improved outcomes with modern care. AB - The outcome of necrotizing soft tissue infections (NSTI) remains unchanged despite advances in care. Reasons cited are changing patterns of causative factors, delays in diagnosis, and inadequate antibiotic coverage and surgical treatment. To document outcomes of NSTI after aggressive management and to identify risk factors of mortality and prolonged hospital stay, we reviewed all our patients with NSTI admitted from January 2000 to January 2002. Causative factors, comorbid conditions, site of NSTI, physiologic parameters, symptoms, diagnostic tests, therapeutic interventions, and outcomes were analyzed. Patients were treated aggressively with antibiotics, admission to ICU, and frequent surgical debridements. Of 46 patients identified, 28 (61%) were admitted in ICU, and eight (17%) died. The patients who died had higher admission white blood cell counts (46 +/- 22 vs 22 +/- 10 x 10(3)/mm3, P = 0.01), higher admission pain score (8 +/- 1 vs 5 +/- 3, P = 0.02), longer intervals from admission to antibiotic administration (16 +/- 20 vs 6 +/- 12 hours, P = 0.02), and fewer surgical debridements (2.6 +/- 1.1 vs 3.6 +/- 1.7, P = 0.04). No independent risk factors of mortality or ICU admission were identified. We concluded that severe local pain and a significantly elevated white blood cell count on admission should alert the physician to the presence of severe infection and prompt the initiation of expeditious aggressive treatment. PMID- 15534962 TI - Time-resolved spectrophotometer for turbid media based on supercontinuum generation in a photonic crystal fiber. AB - We describe an instrument for the time-resolved spectroscopy of turbid media that is based on supercontinuum generation in a photonic crystal fiber. The light injected into the sample consists of subpicosecond pulses that cover 550-1000 nm at 85 MHz at an average power of as much as 40 mW. A spectrometer coupled to a multianode photomultiplier tube is used to detect the light simultaneously in 16 wavelength channels, with a resolution of 5-20 nm/channel, depending on the grating. Time-correlated single-photon counting is used to produce time dispersion curves, which one fits to the diffusion equation to determine absorption and reduced scattering coefficients. We tested the instrument by measuring the time-resolved diffuse reflectance of epoxy phantoms and by performing in vivo measurements on volunteers. The results were similar to those obtained with previous discrete wavelength systems, whereas the full spectrum (610-810 nm) acquisition time was as short as 1 s owing to the parallel acquisition. PMID- 15534963 TI - The effect of surface agitation on ultrasound-mediated gene transfer in vitro. AB - This article reports the effect of surface agitation of culture medium on ultrasound-mediated gene transfection in vitro and its possible mechanisms. The possibility of active induction of bubbles without using contrast microbubbles for effective gene transfer was also demonstrated. Cultured HeLa cells mixed with green fluorescent protein plasmid were exposed to 1.0 MHz ultrasound in 24-well culture plates. Up to 26% transfection efficiency in the survival cell population was achieved in samples exposed to 0.44 MPa ultrasound pulses with the presence of surface agitation. Inertial cavitation and bubble generation were observed throughout the ultrasound exposure. When surface agitation was suppressed by covering the medium surface with a thin membrane, bubble generation and gene transfection were significantly suppressed. Interestingly, transfection efficiency could be partially resumed by adding a small amount of culture medium onto the covering membrane to rebuild the surface agitation and bubble generation. Pressure fluctuation and transient high-pressure loci were found in samples with surface agitation. Numerical simulations of bubble dynamics showed that transient high pressures above the inertial cavitation threshold could generate bubbles, which might be subsequently stabilized at lower pressures by rectified diffusion, and exert strong shear forces that might create transient pores on cell membranes to facilitate gene transfer. PMID- 15534964 TI - [Abstracts from the 65th SIMLII (Italian Society of Occupational Medicine and Industrial Health) National Congress. Messina, Italy, 11-14 September 2002]. PMID- 15534965 TI - Proceedings of the 7th Congress of the International Society for Organ Donation and Procurement and the 4th Congress of the International Transplant Coordinators Society. Warsaw, Poland, November 27-December 1, 2003. PMID- 15534966 TI - Proceedings of the 7th Congress of the International Society for Organ Donation and Procurement and the 4th Congress of the International Transplant Coordinators Society. Warsaw, Poland, November 27-December 1, 2003. PMID- 15534967 TI - Public policy statement on parity in insurance benefit structure between general medical services and services for the evaluation, management and prevention of behavioral health conditions, including addictive disorders. PMID- 15534968 TI - Proceedings and abstracts of the 7th National Conference on Anticoagulant Therapy. San Francisco, California, USA. 8-10 May 2003. PMID- 15534969 TI - Public policy statement on buprenorphine for opiate dependence and withdrawal. PMID- 15534971 TI - Public policy statement on hepatitis C: supplemental information for physicians. PMID- 15534970 TI - Public policy statement on discrimination on the basis of sexual orientation. PMID- 15534972 TI - Public policy statement on the rights and responsibilities of healthcare professionals in the use of opioids for the treatment of pain. PMID- 15534973 TI - Young Scientist Award 2nd ISPRM (International Society of Physical and Rehabilitation Medicine) World Congress. Prague, Czech Republic, May 2003. PMID- 15534974 TI - [Proceedings of the Consensus Conference on Management of Drug-Resistant Partial Epilepsy. 3-4 March 2004]. PMID- 15534975 TI - Advocacy Day preview--meeting with success. PMID- 15534976 TI - Small bowel photography: the jury needs evidence. PMID- 15534977 TI - Effective therapy for advanced gastrointestinal stromal tumors. PMID- 15534979 TI - Itchin' for a diversion. PMID- 15534978 TI - Truth is not determined by a majority vote. PMID- 15534980 TI - Hepatitis C: beyond the liver. PMID- 15534981 TI - Endoscopic blockade to prevent resurfacing of esophageal varices. PMID- 15534982 TI - Leptin signaling and obesity: weight and see. PMID- 15534983 TI - Is it HELLPful to consider the hanging LCHAD in pregnancy-associated liver disease? PMID- 15534984 TI - Suppression of pathogenesis: bacteria lead the way. PMID- 15534985 TI - Terlipressin and albumin for HRS: an advance in therapy? PMID- 15534987 TI - Paul Farmer's grand challenges. Interview by Sue Thomas Hegyvary. PMID- 15534986 TI - Iron and neoplasia: serum transferrin receptor and ferritin in prostate cancer. PMID- 15534988 TI - Breadboard testing of a phase-conjugate engine with an interferometric wave-front sensor and a microelectromechanical systems-based spatial light modulator. AB - Laboratory breadboard results of a high-speed adaptive-optics system are presented. The wave-front sensor for the adaptive-optics system is based on a quadrature interferometer, which directly measures the turbulence-induced phase aberrations. The spatial light modulator used in the phase-conjugate engine was a microelectromechanical systems-based piston-only correction device with 1024 actuators. Laboratory experiments were conducted with this system utilizing Kolmogorov phase screens to simulate atmospheric phase distortions. The adaptive optics system achieved correction speeds in excess of 800 Hz and Strehl ratios greater than 0.5 with the Kolmogorov phase screens. PMID- 15534989 TI - Fabrication and characterization of a three-dimensional feed-horn infrared antenna for an infrared detector. AB - A three-dimensional feed-horn antenna for the 10-microm-wavelength infrared region has been suggested, characterized, and fabricated. It is applied to an infrared detector for efficient collection of infrared radiation and to reduce background noise. The optimum size of the horn antenna was designed for maximum antenna directivity. The three-dimensional feed-horn antenna mold was fabricated by rotating and tilting illumination, whereas the antenna plate was acquired through electroplating. Antenna characteristics were measured by coupling of the antenna with a microbolometer. Measurement results show that the directivity of the antenna is 16.1 dB and the background noise is reduced by a factor of approximately 2 compared with an open-structure infrared detector. PMID- 15534990 TI - Pulse-laser electroholography by use of interference fringe patterns captured by a CCD. AB - Digital holography is combined with a pulse-laser electroholographic system for a real-time three-dimensional display. Owing to the one-dimensional characteristics of the Bragg-regime acousto-optic spatial-light modulator, vertical parallax cannot be generated from the acoustic signal propagating along the fan-shaped beam direction of the incident laser. To obtain a proper interference pattern, we attach a horizontal slit to the confocal lens system for recording the fringe data, significantly reducing the bandwidth of the vertical fringe data. When the bandwidth-reduced fringe data are displayed by use of a pulse-laser electroholographic system, the clarity and the quality of the image are found to be appreciably improved. PMID- 15534991 TI - Development of a diffraction-type optical triangulation sensor. AB - We propose a diffraction-type optical triangulation sensor based on the diffraction theorem and the laser triangulation method. The advantage of the proposed sensor is that it obtains not only the linear displacement of a moving object but also its three angular motion errors. The developed sensor is composed mainly of a laser source, two quadrant detectors, and a reflective diffraction grating. The reflective diffraction grating can reflect the incident laser beam into several diffractive rays, and two quadrant detectors were set up for detecting the position of 0- and + 1-order diffraction rays. According to the optical triangulation relationship between the spatial incident angles of a laser beam and the output coordinates of two quadrant detectors, the displacement and the three angular motion errors of a moving object can be obtained simultaneously. PMID- 15534992 TI - Optical memory effect in a deformed helix ferroelectric liquid crystal. AB - Optical memory in a deformed-helix ferroelectric liquid crystal is proposed by deforming the helix under the application of a square-voltage pulse of known magnitude and frequency. This effect is based on the electromechanical effect of helix deformation due to the electric field. When the interaction between the electric field and the dipole is sufficiently strong, all of the dipoles align along the electric field. In such a situation the interlayer dipole-dipole interaction is strong enough to balance the elastic deformation energy. When the electric field is switched off, the molecules remain in a static, balanced state owing to the dipole-dipole interaction and hence the memory effect. PMID- 15534993 TI - Achromatic hybrid refractive-diffractive lens with extended depth of focus. AB - A method for designing achromatic hybrid refractive-diffractive elements that can produce beams with long focal depths while they preserve the entire aperture for capture of light and high transverse resolution is presented. Its working principle is based on the combination of a diffractive optical element that generates a long range of pseudonondiffractive rays and a refractive lens of opposite dispersion to form an achromatic hybrid lens. A hybrid lens with a fast f-number (f/1) that works in the entire visible wave band (400-700 nm) was designed and fabricated. Simulation results demonstrate a factor-of-10 improvement in depth of focus compared with that of a conventional f/1 lens, with matching 1-microm lateral resolution. Experimental results confirm the effectiveness of the proposed method through demonstration of an achromatic hybrid lens with better than a factor-of-7 improvement in depth of focus and 1 microm transverse resolution. PMID- 15534994 TI - Coupling of light from an LED into a thin light guide by diffractive gratings. AB - Ring-shaped and radial diffractive gratings are designed with rigorous diffraction theory to couple light of a nearly monochromatic LED into a thin planar light guide on the bottom side. The theoretical coupling efficiencies for ring-shaped and radial gratings are 41% and 66%, respectively. Optimized diffractive elements are manufactured with direct electron-beam lithography and reactive-ion-etching into SiO2 substrates. Good agreement between experimental and theoretical results for selected radial gratings is reached. Furthermore, the mass production tests using injection molding are carried out with good replicability. PMID- 15534995 TI - Terahertz imaging system based on a backward-wave oscillator. AB - We present an imaging system designed for use in the terahertz range. As the radiation source a backward-wave oscillator was chosen for its special features such as high output power, good wave-front quality, good stability, and wavelength tunability from 520 to 710 GHz. Detection is achieved with a pyroelectric sensor operated at room temperature. The alignment procedure for the optical elements is described, and several methods to reduce the etalon effect that are inherent in monochromatic sources are discussed. The terahertz spot size in the sample plane is 550 microm (nearly the diffraction limit), and the signal to-noise ratio is 10,000:1; other characteristics were also measured and are presented in detail. A number of preliminary applications are also shown that cover various areas: nondestructive real-time testing for plastic tubes and packaging seals; biological terahertz imaging of fresh, frozen, or freeze-dried samples; paraffin-embedded specimens of cancer tissue; and measurement of the absorption coefficient of water by use of a wedge-shaped cell. PMID- 15534996 TI - Generalization of the Jared and Ennis method of complex transmittance objects for the generation of synthetic discriminant function filters. AB - We present a simple method of constructing synthetic discriminant function filters optimized to take into account the modulation of liquid-crystal devices. This relaxation algorithm, a generalization of the Jared and Ennis method, is an iterative method that includes arbitrary modulations for both scene and filter, extending the problem to the complex plane. Simulated and experimental results obtained in a VanderLugt correlator are presented for a two-class recognition problem. The optimal number of images needed to describe an object in a filter generated in this way is discussed, and the influence of the spatial light modulation resolution on the correlation is studied. PMID- 15534997 TI - Two-dimensional phase unwrapping with a multichannel least-mean-square algorithm. AB - This paper presents a computationally efficient two-dimensional phase-unwrapping method based on a multichannel least-mean-square algorithm. The performance of the proposed method is evaluated by applying phase unwrapping to several simulated very noisy images and to a genuine noisy interferometrical image taken from a five-step phase-shift interferogram obtained from a surface plasmon resonance imaging biosensing experiment. The results confirm that the proposed method is more widely applicable, more computationally efficient, and more robust in the presence of noise than the representative methods presented in this paper. PMID- 15534998 TI - Light scattering on oceanic turbulence. AB - Turbulent inhomogeneities of fluid flow have the effect of scattering light in near-forward angles, thus providing an opportunity to use optics to quantify turbulence. Here we report measurements of the volume-scattering function in the range of 10(-7) to 10(-3) rad using a wave-front sensing technique. The total scattering coefficient b, due to scattering on turbulent inhomogeneities, is between 1 and 10 m(-1) under typical oceanographic conditions. The numerical calculations of turbulent volume-scattering functions compare well with the laboratory measurement. These results suggest that optical measurements at small angles are affected by turbulence-related scattering, and their effects can be well modeled with numerical calculations. PMID- 15534999 TI - Quantitative imaging through a spectrograph. 1. Principles and theory. AB - Laser-based optical diagnostics, such as planar laser-induced fluorescence and, especially, Raman imaging, often require selective spectral filtering. We advocate the use of an imaging spectrograph with a broad entrance slit as a spectral filter for two-dimensional imaging. A spectrograph in this mode of operation produces output that is a convolution of the spatial and spectral information that is present in the incident light. We describe an analytical deconvolution procedure, based on Bayesian statistics, that retrieves the spatial information while it avoids excessive noise blowup. The method permits direct imaging through a spectrograph, even under broadband illumination. We introduce the formalism and discuss the underlying assumptions. The performance of the procedure is demonstrated on an artificial but pathological example. In a companion paper [Appl. Opt. 43, 5682-5690 (2004)] the method is applied to the practical case of fuel equivalence ratio Raman imaging in a combustible methane air mixture. PMID- 15535000 TI - Quantitative imaging through a spectrograph. 2. Stoichiometry mapping by Raman scattering. AB - The Bayesian deconvolution algorithm described in a preceding paper [Appl. Opt. 43, 5669-5681 (2004)] is applied to measurement of the two-dimensional stoichiometry field in a combustible methane-air mixture by Raman imaging through a spectrograph. Stoichiometry (fuel equivalence ratio) is derived from the number density fields of methane and nitrogen, with a signal-to-noise ratio of approximately 10 in a 600-laser-shot average. Prospects for single-shot Raman imaging are discussed. PMID- 15535001 TI - Intrinsic properties of the optical coupling between axisymmetric Gaussian beams. AB - On the basis of the overlap integral method, an approximate analytical model is derived to estimate the coupled optical power between axisymmetric Gaussian beams when transverse, axial, and angular misalignments simultaneously exist in three dimensions. Seven optical properties are derived from a detailed analysis of the model. Because the model is an approximate analytical solution to the overlap integral method, the existence of each property is also investigated by a numerical solution. Results show that all seven properties are intrinsic to the optical coupling phenomenon between Gaussian beams. Because numerous single-mode device-to-fiber coupling systems can be well described by use of Gaussian beams, the seven properties provide a solid basis to develop model-based algorithms for single-mode device-to-fiber alignment automation. PMID- 15535002 TI - Experimental study of bent multimode optical waveguides. AB - Bend losses of bent multimode waveguides have been experimentally studied. The results experimentally verify a simple analytical expression based on ray optics that can be used to determine bend losses in bent multimode waveguides. We have also shown that bend-induced losses in such multimode optical waveguides are wavelength independent. PMID- 15535003 TI - Spectral airglow temperature imager (SATI): a ground-based instrument for the monitoring of mesosphere temperature. AB - The spectral airglow temperature imager is a two-channel, Fabry-Perot spectrometer with an annular field of view and a cooled CCD detector. The detected fringe pattern contains spectral information in the radial direction and azimuthal spatial information from the annular field of view. The instrument measures the rotational temperature from the O2 atmospheric (0,1) nightglow emission layer at 94 km and from the Q branch of the OH Meinel (6,2) band emission layer at 87 km. The method for temperature derivation is based on the temperature dependence of the line-emission rates. This dependence allows a determination of the temperature by a least-squares fit of the measured spectrum to a set of synthetic spectra, an approach that minimizes the effect of noise from the sky background and the detector. The spectral airglow temperature imager was developed to meet a need for monitoring the role of the mesosphere in climate variability through long-term observation of the mean temperature and the gravity waves from a single station, as well as large-scale wave perturbations through the use of multiple stations. PMID- 15535004 TI - [Immigrant population and communicable diseases]. PMID- 15535005 TI - [Methodology for the implementation of e-Health services for chronic patient monitoring and control]. AB - A methodology is presented for a smooth, orderly implementation of specific e Health services for monitoring chronic patients outside of the hospital setting. Identified as a stage-gate model for the management of the overall implementation process, this methodology is presented formally structured into three steps: a) exploratory examination (pilot project stage); 2) in-depth evaluation (clinical trial stage); and 3) deployment (guided use stage). In the first stage, controlled by the R+D team, the predominant criiteria are the functionality and usability of the technologies involved. In the second stage, controlled by an associated Health Technology Evaluation Agency, the predominant criterion is the scientific aspect related to the results obtained in the clinical testing. The third stage is controlled through decisions made by the health administrations as to the implementation of new technologies and the financing thereof. A description is provided as to the requirements of the technological platform designed to serve its the medium for the projects and tests from stage 1 and 2. As an example of what is done in stage 2, a description is given of a trial related to hypertension. PMID- 15535006 TI - [Occupational accident injuries in Spain: a comparison among Autonomous Communities in 1989, 1993 and 2000]. AB - BACKGROUND: Despite the incidence of occupational accident injuries in Spain being one of the highest in the European Union, this health problem has been researched very little to date. This study was aimed at comparing the incidence of fatal and non-fatal occupational accident injuries by Autonomous Communities in 1989, 1993 and 2000. METHODS: The occupational accident injury data was obtained from the Ministry of Labor and Social Affairs, and the denominators from the Spanish National Institute of Statistics. The incidence was calculated for fatal and non-fatal occupational accident injuries, adjusting separately by sex, age, business activity and type of contract, taking the Spanish average as a reference. RESULTS: Catalunya and the Baleric Islands showed the highest relative risks of non-fatal occupational accident injuries. Regarding fatal occupational accident injuries, the Asturias and the Basque Autonomous Communities were the Communities where the relative risk was highest. Madrid and Catalunya, on the contrary, having showed the lowest RR for fatal injuries. CONCLUSIONS: The excessive risk of occupational accident injuries found in some Autonomous Communities can be explained, after partially ruling out the differences in the business activities and type of contract, either in terms of differences in the reporting of the occupational accident injuries and/or of the population at risk, or rather, in terms of differences in the working conditions at the companies in these Autonomous communities, which is the most plausible hypothesis. These differences must be studied further in greater detail. PMID- 15535007 TI - [Prevalence of tuberculosis infection among immigrants in the Toledo Health District, Spain]. AB - BACKGROUND: The rise in the frequency of tuberculosis is due to several factors: resistance to treatment, human immunodeficiency virus infection, unemployment, poverty and migratory movements. This study is aimed at ascertaining the prevalence of tuberculosis infection among the immigrant population in the Toledo Health District. METHODS: Descriptive, cross-sectional, multi-center study conducted at the Primary Care level. The immigrants were attracted and recruited on the based of their health cards within the December 2002-September 2003 period. Mantoux intradermal reaction test was performed with 0.1 ml (2UT) of PPD RT-23, following informed consent. Other variables were collected: age, sex, weight, country of origin, work situation, BCG vaccination, number of individuals living with them, length of time residing in Europe. RESULTS: A total of 344 immigrants were included in the study, averaging 28.8 +/- 12.7 years of age. Fifty percent were females (172). They had been living in Europe for an average of 3.4 +/- 3.7 years. A total 78.8% (271) of the immigrants completed the study. The intradermal reaction was positive in 75 of them [27.7% (CI 95%: 23-32.5)]. Those immigrants showing a positive Mantoux were older (age 32.5 +/- 8.5 years) than those showing a negative result (age 27.7 +/- 13.8 years), p=0.006. Originally being from the sub-Saharan area increased the probability of showing a positive intradermal reaction. CONCLUSIONS: The immigrant population of the Toledo Health District shows a high prevalence of tuberculosis infection, which is higher than the general Spanish population. Differences exist depending upon the country of origin. These results indicate the importance of considering the Mantoux test among the preventive activities addressed toward this immigrant population. PMID- 15535009 TI - [Evaluation of the SF-36 health index applied to methadone maintenance program users. Reference values for the Basque Autonomous Community, Spain]. AB - BACKGROUND: The questionnaires that measure perceived health are used in clinical practice to determine the impact of illnesses and the benefits of treatments. Population-based norms have been proposed to increase their interpretability. The aim of this paper was to obtain reference values for users of Methadone Maintenance Programs (MMP) in the Basque Autonomous Community (BAC). METHODS: Cross-sectional study carried out in 2000 on a stratified random sample of users included in MMP of BAC (n=726). SF-36 Health Survey was used; central trend, dispersion and percentile data were estimated for each of the SF-36 dimension scores to generate reference values according the dwell time in the methadone program. RESULTS: All Cronbach's alpha coefficients were higher than 0.7, and the percentage or non-responds were lower 1%. The MMP users had lower punctuations in all dimensions of SF-36 than population values of reference (p<0.001). The health status of the participants was worse among those than they began the treatment that among those that took more than six months in the MMP; the differences in all SF-36 dimensions scores were statistically significant (p<0.001). CONCLUSIONS: The SF-36 as a measure of health status in MMP population shows a high internal consistence, and is able to discriminate patients with different stages of treatment evolution. Results presented should be considered the population-based norms for monitoring the improvement progression or response to treatment in patients included in MMP. PMID- 15535008 TI - [Factors related to experiencing respiratory failure in cases of opiate overdose for which care was provided in an open setting. Barcelona, Spain]. AB - BACKGROUND: To determine the factors associated with respiratory arrest in opiate overdoses (coma, pupillary miosis, respiratory depression, and response to naloxone) among injecting drug users in the Can Tunis quarter of Barcelona. METHODS: We ran a transversal observational study where all overdoses assisted between March, 2001 and June, 2002. After overdose treatment, data were collected using a standard questionnaire, including: patients' sociodemographic data, opiate and other substances' use prior to overdose, clinical signs and symptoms presented, and medical intervention received, by way of a standardised questionnaire. Logistic regression was used as a tool for analysis. RESULTS: Of 222 opiate overdose cases, 60.8% showed respiratory arrest. Of all risk factors tested, only prior abstinence heroin abstinence for 2 weeks or longer days (OR=1.893; p=0.04), and no previous consumption of benzodiazepines (OR:0.462; p=0.017), proved to have a statistically significant association with suffering a respiratory arrest. Concomitant use of alcohol, cocaine or methadone appeared not associated with suffering respiratory arrest in opiate overdose. CONCLUSIONS: The main risk factor for respiratory arrest in opiate overdoses was a prior abstinence period of more than 2 weeks. Benzodiazepines use was associated with absence of respiratory arrest in overdose cases. Alcohol or methadone use, as well as the use of larger quantities of heroin, was not associated with suffering respiratory arrest in opiate overdoses. A study of other factors, not included in this study, and that could interfere with our results, should be considered for their possible relationship to benzodiazepine use as well as to absence of respiratory arrest in overdose cases. PMID- 15535010 TI - [Screening of microfilariasis in blood (Loa Loa) among the immigrant population in endemic areas]. AB - BACKGROUND: Given the increasing flux of immigrant population from high-risk areas to our country, the need of screening for loasis arises, despite systematic screening being a debated and poorly evaluated practice to which there is no protocol. This study is aimed at identifying the population to which loasis screening would be most appropriate, by drawing a comparison among four alternatives. METHODS: Case and control group study, involving 30 cases (all those who came to our unit for treatment and who tested positive for Loa Loa in a microfilariae in the blood detection test) and 90 control cases (three controls per case from among the subjects having tested negative for microfiliariae in the blood chosen at random without any pairing criteria). RESULTS: Of the 1,638 subjects on whom the microfilariae blood test was performed, 30 tested positive (1.8%; 1.2-2.6%). Of these 30 cases of loasis, 76.7% (23; 57.7-90.1%) had eosinophilia (OR 8.8; 3.3-23.1; p<0.0001) and 30.0% (9; 14.7-49.4%) compatible clinical symptoms (OR 2.8; 1.0-7.5; p=0.04). If we were to apply the screening test to the entire immigrant population coming from endemic areas, we would have to perform 54.6 tests to detect one case. If we were to perform the test on patients showing eosinophilia and/or compatible clinical symptoms, we would have to perform a smaller number of tests for every case detected (NNS=29; IC=21-48), but there would be 16.7% (5; 5.7-34.7%) false negatives. CONCLUSIONS: Conducting a screening test with determination of microfiliariae in the blood on the immigrant population coming from Central and West Africa, independently of the presence of eosinophilia or compatible clinical symptoms, would be indicated, provided that the necessary resources are available. PMID- 15535012 TI - The 2004 state elections: let me be perfectly Blunt! PMID- 15535011 TI - [Trends in the utilization of antidepressant drugs in the Rioja and Zamora health districts throughout the 1997-2001 period]. AB - BACKGROUND: Due to the high increase in the utilization of antidepressant drugs and the change in the prescription profile within this group, this study is aimed at discovering the evolution of the utilization of antidepressant drugs (NO6A subgroup) outside of the hospital setting in the Rioja and Zamora health districts throughout the 1997-2001 period and to evaluate the impact of the new drugs. METHODS: A study was made of the utilization of drugs included in the NO6A treatment subgroup under the Anatomical Therapeutic Chemical (ATC) classification. The data on the utilization of each specialty was taken from the pharmacy management (SIFAR) software applications and has been stated for each active ingredient in Daily Dose per Inhabitant (DDI). RESULTS: In the period under study, there was an increase in the total utilization of antidepressants in the two areas (Zamora 55% and Rioja 93%) due mainly to the increase in selective serotonin reuptake inhibitors (SSRI) (Zamora 80% and Rioja 143%) and in the new heterocyclic antidepressants (Zamora 82% and Rioja 78%). CONCLUSIONS: There has been an increase in the utilization of antidepressants, all of which has taken place in conjunction with a change in the prescription profile. There is a clear shift toward the utilization of the SSRI's and some heterocyclic antidepressants (venlafaxine, trazodone, reboxetine), the utilization of tricyclic antidepressants and IMAO's having decreased. PMID- 15535013 TI - Workable medical malpractice solutions brought forth by experience. PMID- 15535014 TI - A prescription for Missouri. PMID- 15535015 TI - Elizabeth Blackwell, MD--the first female physician in the United States. PMID- 15535016 TI - While you are busy practicing medicine. PMID- 15535017 TI - Future of American medicine 2005-2010: questions, answers & opinions. PMID- 15535018 TI - Medical education in the twenty-first century: the dean's perspective. PMID- 15535019 TI - Unique medical education and curricular reform at the Kansas City University of Medicine and Biosciences. PMID- 15535020 TI - Defibrillators require training. PMID- 15535021 TI - Thank you to Missouri physicians. PMID- 15535022 TI - Sutton's law. PMID- 15535023 TI - Biomedical education in the 21st century. AB - The extraordinary discoveries of 20th century medicine and technology have created a scientific renaissance. This explosion of new knowledge and the tantalizing potential it holds for altering the course of human health and disease will change the practice of medicine and require the education of a new generation of translational/clinical scientists and physician-scientists as well as an accelerated evolution of the teaching paradigms for the training of physicians. PMID- 15535024 TI - The clinical experience in medical education: past, present, future. AB - In the nineteenth century, clinical education in the United States was entirely didactic. Medical students attended lectures all day and were expected to commit the many details to rote memory. In the modern era, the clinical clerkship transformed students from passive observers to active participants in the learning process. In addition, the internship and residency provided learners the opportunity to assume responsibility in patient care. The strength of this clinical experience depends not only on the will of medical faculties but on the quality of the hospital learning environment. PMID- 15535025 TI - The Washington University Medical Scientist Training Program. AB - Washington University's Medical Scientist Training Program (MSTP) is the largest and among the most successful programs in the country for training physician scientists. Due to the evolution of biomedicine and as a result of recent initiatives at the National Institutes of Health (NIH), exciting new opportunities are arising for physician-scientists. Interdisciplinary research will be increasingly required to tackle challenging scientific and medical problems, and MSTP students able to negotiate between the physical, biological and medical sciences will be poised to take advantage of this. PMID- 15535026 TI - Proteomics, genomics and the future of medical education. AB - The completion of the human genome project in 2003 ushered in the era of genomics, the systematic study of our DNA sequence. Proteomics, the study of the full complement of proteins present in a cell, is a natural extension of genomics. Together, the information obtainable through genomics and proteomics has tremendous potential to change clinical practice. The application of such information to medical diagnosis and treatment will require significant changes in the training of physicians. All students and physicians in training will need to acquire enough knowledge of the underlying science, including medical genetics, epidemiology, bioinformatics and statistics, so they will intuitively understand the technology and recognize the strengths and limitations of genomic/proteomic tests. Because genomic or proteomic testing may yield extensive information about a person's genetic makeup and disease risks, consideration will need to be given throughout the medical curriculum to the ethical issues raised by the application of this new technology to the diagnosis and treatment of patients. PMID- 15535027 TI - Integrating social and ethics-based aspects of medicine into physician education. AB - Ethics plays an important, formative role in medical education, stimulating the transformation of a physiologist-scientist into a compassionate and competent clinician. Our clinical ethics program challenges students to observe and recognize inherent value conflicts and understand how such issues must be resolved in their day-to-day work. Advances in biomedical research that hold potential to enhance human health and treat disease are provoking controversial questions of current practices in medical ethics and spawning new, unprecedented ethical dilemmas. PMID- 15535028 TI - Stroke: a cardiologist's perspective. AB - This review is an attempt to give the primary care physician an overview of the risk factors for stroke in 2004. In addition, the available treatments both medical and interventional were reviewed. PMID- 15535029 TI - Comparing practice differences of residency graduates: implications for curricular change. AB - OBJECTIVE: To document and assess differences in the practice experiences and practice patterns of graduates from two similar sized but regionally separated community-based internal medicine residency programs. RESULTS: Sixty percent responded to the survey. Responses from graduates practicing general internal medicine were used in the analyses (Wichita n = 20; Baystate n = 23). Only graduates from Wichita were in solo practice (15%). Baystate graduates had a significantly higher percentage of HMO patients (32% vs. 17%, p < .05). A higher percentage of Kansas respondents worked in a community with a population of less than 50,000 (55% vs. 26%, p = .052). Of 28 components of a residency curriculum, 7 were considered significantly more important to daily practice by Kansas respondents than by Baystate respondents: Critical Care, Dermatology, Gastroenterology, Nephrology, Neurology, Occupational Medicine, and Rheumatology. Kansas respondents performed on average significantly more procedures than Massachusetts respondents in the last year in 6 of 16 procedures: bone marrow biopsy, exercise stress tests, flexible sigmoidoscopy, liquid nitrogen, skin biopsy, and thoracentesis. The procedures of skin biopsy, stress testing, and the curricula of dermatology, nephrology, neurology and rheumatology remained significantly different when controlled for the size of the community population (<50,000). Wichita graduates scored higher on the practice intensity measure than Baystate graduates. CONCLUSIONS: We have documented differences in the importance of particular curricula, procedures, and practice intensity likely related to the community population in which residency graduates practice. Understanding the needs of graduates and incorporating this information into existing rotations or new initiatives is integral to the ongoing development of residency curricula. PMID- 15535030 TI - Toll-like receptors in rat brains injured by hypoxic-ischaemia or exposed to staphylococcal alpha-toxin. AB - Some data suggest that the central nervous system (CNS) is the main target of Staphylococcus alpha-toxin. Since this pathogen cannot penetrate the blood-brain barrier (BBB), the exact mechanism by which alpha-toxin affects the CNS remains unclear. Recent studies on the role of the innate immune system have shed light on how bacterial infections initiate inflammatory responses within the CNS. The aim of this study was to investigate the immunoexpression of Toll-like receptors (TLR 2, TLR 4) in brains of young rats systemically exposed to Staphylococcus alpha-toxin or injured by neonatal hypoxia-ischaemia. The study was carried out on 6-week-old Wistar rats. A group of 6-week-old rats with severe brain injury caused by neonatal hypoxia-ischaemia was also studied separately. In all control rats, the immunoexpression of TLR 2 and TLR 4 was not detected. However, the expression of both TLRs was evident in all brains injured by HI or exposed to alpha-toxin. The immunoexpression was localised in the wall of the small brain vessels, cells of ependyma and leptomeninges. In such vessels the spectrum of ultrastructural lesions was found. The presence of TLR4 detected in the nerve cells of the subcortical gray matter of the brain is particularly of interest, but requires further studies. The presence of TLR 4 antigen in the nerve cells of the subcortical gray matter is particularly of interest. In conclusion, the results show that brain microvessels through TLRs may participate in the immune response of brain affected by bacterial infection as well as injured by non infection insults. PMID- 15535031 TI - Cerebrospinal fluid free-radical peroxidation products and cognitive functioning patterns differentiate varieties of normal pressure hydrocephalus. AB - The aims of the study were as follows: first, to verify the hypothesis that free radical peroxidation may be one of the factors implicated in pathophysiology of normal pressure hydrocephalus (NPH) and, second, to find out whether these biochemical characteristics together with neuropsychological cognitive deficits can differentiate between various types of NPH. This provides prognostic criteria for selection of patients for shunt surgery. Lipid peroxidation was measured in terms of thiobarbituric acid-reactive material (TBAR) and protein sulphydryl (SH) groups were measured as CSF content. Cognitive deficits were assessed using a number of neuropsychological tests. In the sample of NPH patients (n = 24), three categories were distinguished using these criteria: idiopathic active hydrocephalus (A), arrested hydrocephalus (AH), and post-traumatic hydrocephalus (PT). TBAR levels for NPH patients were higher than that of controls without CNS pathology (n = 2). Moreover, NPH patients had increased levels of total and soluble protein groups, and decreased levels of protein SH groups, which suggests the occurrence of processes that activate peroxidation of free radicals in normal pressure hydrocephalus. Levels of these indicators varied across NPH types. Two categories of NPH patients, with active (A) or posttraumatic (PT) hydrocephalus differed significantly from the controls (C)--their TBAR levels were 0.58, 0.56 and 0.28 nmol/mg protein, respectively; soluble SH levels: 41.5; 58.15 and 11.3 nmol/mg protein, and protein SH levels: 34.3, 21.8 and 57.5 nmol/mg protein. In PT group, many individual differences were noticed. These findings seem promising because the studied biochemical indicators may serve as additional diagnostic criteria for selection of NPH patients for shunting. PMID- 15535032 TI - Limbic neuropathology in idiopathic Parkinson's disease with concomitant dementia. AB - To study pathological background of dementia in idiopathic Parkinson's disease (PD), 41 autopsy brains (31 cases with and 10 cases without dementia) were investigated. The severity of degenerative changes was evaluated in selected limbic regions (trans- and entorhinal cortex, hippocampus, and amygdala). The densities of Lewy bodies (LBs), Lewy neurites (LNs), neurofibrillary tangles (NFTs), and amyloid neuritic plaques (NPs) were determined on immunohistochemically stained sections using antibodies against alpha-synuclein, tau-protein, and amyloid-beta. Precisely defined modern criteria for selecting study cohort (Newcastle, CERAD and Braak et al.) ensured homogeneity of the study sample and reliability of the results. Comparisons between the cases of Parkinson's disease with dementia (PDD) and those without (PD-only) revealed that the former were characterised by significantly higher densities of LBs and LNs in transentorhinal and entorhinal cortices as well as in the CA2-3 region of the hippocampus and cortical complex of amygdala. In the PDD sub-set we found statistically significant correlation of LBs with LNs counts in CA2-3 region of hippocampus as well as of LBs counts in transentorhinal cortex with LNs counts in CA2-3 hippocampal region. The relationship was also observed between LBs counts in CA2-3 region of the hippocampus and LNs counts in cortical complex of amygdala. Our studies suggest that dementia in PD may be associated with the presence of degenerative changes of PD-type in leading limbic structures, without co-existent Alzheimer's disease (AD). They also imply that LBs and LNs may appear to be morphological hallmarks of the pathological process associated with dementia in PD. LBs and LNs distribution pattern and correlations of LBs with LNs counts in limbic regions observed in our study suggest the cumulative patomechanism of changes dependent on transsynaptic alpha-syn pathology and indicate the spread of the pathological process via axonal transport. The coexistence of the small number of changes of AD-type may exacerbate cognitive deficits in PDD. PMID- 15535033 TI - Oxidative damage to proteins in the spinal cord in amyotrophic lateral sclerosis (ALS). AB - Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease, which has been linked to the generation of free radicals and oxidative stress. Oxidative damage to spinal cord proteins is suggested to be a contributory factor to neuronal death in ALS. Since proteins are the major targets for free radicals and the so-called "reactive species", therefore the objective of our study was to identify oxidatively damaged spinal cord proteins. The material consisted of spinal cords of 8 sporadic ALS cases and 5 controls. We estimated the level of protein carbonyl moieties, which react quantitatively with 2,4 dinitrophenylhydrazine (DNPH). Afterwards proteins were separated by SDS polyacrylamide gel electrophoresis and the protein bound DNPH moieties were detected immunochemically. We also morphologically examined spinal cords after immune staining against DNPH. The protein carbonyl content of the ALS spinal cords significantly increased in all examined cases. In most ALS patients, proteins with 125 kDa, 70 kDa and 36kDa were highly oxidized. The 70-kDa protein was identified immunochemically to be neurofilament 68. The morphological examination of ALS spinal cords indicated a pronounced anti-DNPH immune reaction in neurones of the anterior horns; the reaction in the posterior horns was less intense. Microglia in the white matter was immunoreactive; astroglia was DNPH negative. Although the exact mechanism by which reactive oxygen species induce motor neurones to die is not known yet, the presented data indicate that they affect spinal cord cellular proteins, including neurofilament 68. In this study, we successfully examined the neurochemical features accompanying motor neuron injury in ALS, and the results may help to develop a rationale anti-oxidative neuroprotective strategy. PMID- 15535034 TI - Degeneration of microglial cells in frontal and temporal lobes of chronic schizophrenics. AB - Schizophrenia is a social disease that occurs in 0.5-1% of the population. It shows a high variability in both clinical picture and theory of its pathogenesis. Its clinical manifestations are accompanied by biochemical, immunological and structural changes. A pivotal role in the development of psychotic disorders is attributed to the impaired limbic system. The aim of this study was to find out whether, and if so, to what extent immunocompetent cells of the central nervous system (microglia) are involved in the process of degeneration occuring in these structures. The study was carried out on 12 brains of female chronic schizophrenics. Sections of frontal and temporal cortex were subjected to ultrastructural as well as histochemical and immunohistochemical examinations by light microscopy. In the structures under study, a large number of ramified microglial cells showing on their surface the expression of the major histocompatibility complex class II (MHC II) was observed. Most cells showed degenerative traits (cytoplasm shrinkage, thinning, shortening and fragmentation of their processes) up to apoptotic changes. Perivascular microglia displayed the lowest intensity of degenerative changes. Ultrastructurally, some damaged microglial cells contained phagosomes and/or degenerated mitochondria. Most abnormal microglia showed morphological signs of the former normal function of immunocompetent and phagocytosing cells. Degeneration of microgial cells, resulting most likely from the primary impairment of the neuron-glia communication that damages their immunocompetent function, may lead to the exacerbation of structural damage and psychotic symptoms. Treatment of chronic schizophrenics should involve the supply of agents to prevent degeneration of microglia and/or long-term immunotherapy. PMID- 15535035 TI - Morphometry of synaptophysin immunoreactive ganglion cells in Auerbach plexus in patients with colorectal cancer. Is this a new prognostic factor? AB - The aim of our study was to estimate morphometric parameters of synaptophysin (Syn-38) immunoreactive ganglion cells in colorectal cancer (within and at various distances from neoplastic infiltration) in postoperative material from 60 patients. We analysed the intensity of Syn-38 expression in Auerbach ganglion neurones, mean number of these cells in the ganglion, and their longitudinal and transverse diameters. The results showed a statistically significant reduction in the number of neurones in intramural ganglia of the large intestine located in neoplastic infiltration and in its close proximity. The size of ganglion cells was directly proportional to the distance from cancer infiltration and inversely proportional to Syn-38 content, which may be explained by degenerative changes and dysfunction of these cells. This correlation was significant in the case of cells with the cytoplasmatic Syn-38 immunoreactivity pattern, but did not refer to the cells with perimembranous pattern, which seemed to be undamaged. Morphometric analysis of synaptophysin immunoreactive ganglion cells in Auerbach plexus in colorectal cancer may be a new useful marker for the description of changes in the intestinal nervous system as well as a prognostic factor for colorectal cancer. PMID- 15535036 TI - Brachial plexus tumours--own experience in diagnostics and surgical treatment. AB - Brachial plexus tumours are rather rare peripheral nerve tumours. In this study, we have presented the results of surgical treatment of cases with neoplasmatic process in the brachial plexus structures. Clinical material consisted of 9 patients (8 women, 34-72 years old; and 1 boy, 7 years old) on which the following procedures were performed: a) excision of the tumour with immediate nerve grafting of the sural nerve, b) extirpation of the tumour, c) excision of the tumour and direct suture reconstruction, d) brachial plexus neurolysis, e) tumour biopsy. We evaluated the pre- and post-operative motor and sensory deficit and established the three point scale of deficit intensity from + to +++. We diagnosed the following tumours in the clinical material: neurofibroma (diagnosed in 4 cases), plexiform neurofibroma (diagnosed in 2 cases), neurilemmoma (diagnosed in 1 case), and malignant tumours (diagnosed in 2 cases), which were found at the plexus area; they were metastatic or spreading continuously from the surrounding tissue. After surgical treatment, the sensory and motor function improved significantly in 4 cases (44,4%). The surgical treatment results depended on the size, localization and histological pattern of the tumour. PMID- 15535037 TI - Adrenal Schwannoma. Report of two cases. AB - Adrenal tumours are frequently incidental discoveries, and their therapy is a subject of controversial discussions. Herein we describe two cases of adrenal schwannoma discovered during autopsy. The accidental observation of two benign schwannomas of the adrenal gland on asymptomatic patients led to the hypothesis that the real frequency of this type of lesion was underestimated. Furthermore, the adrenal origin of the superior retro-peritoneum schwannomas represents a diagnostic hypothesis to be considered when the original structure of the neoplasia can't be established. PMID- 15535038 TI - Alzheimer lesions after ischemia-reperfusion brain injury. AB - For now the best-established and accepted theory in Alzheimer's disease (AD) etiology by most scientists is the "amyloid theory", as the main molecular factor of neurodegeneration in AD. We critically review these observations and highlight inconsistencies between the predictions of the "amyloid hypothesis" and the published data. The research of neurobiology of AD, now more than ever, needs an infusion of new concepts. Handful researchers now recognize brain ischemia as a prominent feature in AD and a potential target for therapy aimed at treatment and prevention of disease. The "ischemia-reperfusion theory" was primarily aimed at stimulating study and redirecting the focus of investigations towards ischemic cellular mechanisms of AD. To accommodate the recent progress of study in AD there is a need to synthesize all the divergent pieces of data into a coherent story. This review provides a synopsis of current information about ischemic cellular and molecular mediators involved in Alzheimer's neuropathology as well as interactions between these mediators that influence pathology. In this paper, current knowledge on the close relation between vascular disease factors and Alzheimer's type dementia will be reviewed. We will summarize the data with a special focus on Alzheimer lesions in experimental brain ischemia. Taken all together, evidence presented in this review suggests a scheme for Alzheimer's pathogenesis with ischemia playing a crucial role in influencing and linking beta amyloid deposition to neuronal damage and clinical disease. PMID- 15535039 TI - Mutations in the neurofilament light chain gene (NEFL)--a study of a possible pathogenous effect. AB - Neurofilaments (NFs) have been shown to be involved in the molecular pathology of numerous neurode-generative human disorders. Recently a set of mutations in the neurofilament light gene (NF-L) was reported in patients suffering from axonal and demyelinating forms of Charcot-Marie-Tooth disease (CMT1 and CMT2). Although a few of the NEFL gene sequence variants have been shown to be rather pathogenous mutations than harmless polymorphisms, the status of some of these variants remains unclear. The aim of this study was to analyse a potential pathogenous effect of the mutations in the NEFL gene identified in CMT affected patients. PMID- 15535040 TI - [Comparative studies on activities of antimicrobial agents against causative organisms isolated from patients with urinary tract infections (2002). III. Secular changes in susceptibility]. AB - The bacteria (Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa) isolated from patients diagnosed as urinary tract infections (UTIs) in 13 institutions in Japan were supplied between August 2002 and July 2003. The susceptibilities of these bacteria to various antimicrobial agents were examined. The bacteria were divided into 2 groups consisting of uncomplicated UTIs and complicated UTIs (with and without indwelling catheter) based on their isolation origins. The results were compared with those obtained between 1993 and 2001. The drug sensitivity of S. aureus in this year was similar to those in up to the previous year and S. aureus showed the best susceptibility to vancomycin. The drug sensitivity of E. faecalis in this year also was similar to those in up to the previous year. The drug sensitivity of E. coli in this year was generally good except penicillins and was similar to those in up to the previous year. Among cephems, cefozopran (CZOP) and cefpirome (CPR) showed the highest potency (MIC90: < or = 0.125 microg/mL). An antibacterial activity of cefotiam (CTM) was similar to it in 10 years ago and was fine (MIC90: < or = 1 microg/mL). The sensitivity of E. coli to carbapenems and carumonam (CRMN) also was good like to CZOP. However, the sensitivity of the complicated UTIs group to quinolones decreased after 2000 and was suggested to develop the resistance to the drug. The drug sensitivity of Klebsiella spp. in this year also was similar to those in up to the previous year. The bacteria showed good susceptibility (MIC: < or = 0.125 microg/mL) to cefmenoxime (CMX), CPR, cefixime (CFIX), flomoxef (FMOX), and CZOP among cephems. The drug sensitivity of P. aeruginosa was generally low. Most of the bacteria were little sensitive to cephems except CZOP and ceftazidime (CAZ). The sensitive bacteria to CZOP and ceftazidime (CAZ) were observed to be 26.8% (15/56 strains) and 39.3% (22/56 strains) in complicated UTIs group, respectively. The sensitivity profile of P. aeruginosa to the other tested drugs was not much different from that in up to the previous year. However, the sensitivity of the bacteria to carbapenems tended to decrease after 2000, and the low sensitive strains (MIC: > or = 256 microg/mL) were detected at 22.2% (2/9 strains) in the uncomplicated UTIs group and 3.6% (2/56 strains) in the complicated UTIs group. PMID- 15535041 TI - [Effect of pazufloxacin mesilate, a new quinolone antibacterial agent, for intravenous use on QT interval]. AB - The potential for QT interval prolongation of pazufloxacin mesilate (PZFX mesilate), a new quinolone antibacterial agent for intravenous use, was investigated by in vitro and in vivo electrophysiology studies. Following results were obtained. In vitro electrophysiology study using guinea pig papillary muscles: PZFX mesilate (30-300 microM) had no effects on resting membrane potential (RMP), action potential amplitude (APA) and action potential duration (APD). Reference quinolones, sparfloxacin (3-30 microM) and moxifloxacin (10-100 microM), had no effects on RMP and APA, but significantly prolonged APD at more than 3 and 10 microM, respectively, while ciprofloxacin (10-100 microM) had no effect on each parameter. In vivo electrophysiology study using anesthetized dogs: PZFX mesilate had no effects on electrocardiograph parameter (PR interval, QRS interval, QT interval and QTc) after intravenous administration of 3-30 mg/kg. These results suggest that PZFX mesilate has low potential for QT interval prolongation. PMID- 15535042 TI - [Development of software for data analysis by therapeutic drug monitoring of teicoplanin, a glycopeptide antibiotic]. AB - We developed a new software named TEICTDM based on the Bayesian estimation utilized in the therapeutic drug monitoring (TDM) of teicoplanin, a glycopeptide antibiotic, for the estimation of individual pharmacokinetic parameters. Therefore, it is necessary to input more than one plasma concentration(s) determined in individual patient. Individual pharmacokinetic parameters were calculated by a least squares methods, MULTI2 (BAYES). Two-compartment model was applied to determine individual pharmacokinetic parameters in male healthy volunteers in Japan, and the relationship between clearance of teicoplanin and creatinine clearance in adult patients with various degrees of renal impairment in Europe was used. A series of work from data input to graph drawing or printing of results could efficiently carried out with the best of use of this software, suggesting that this software is now available in clinical practice. PMID- 15535043 TI - [Antifungal prophylaxis in allogenic hematopoietic stem-cell transplant recipients]. PMID- 15535044 TI - Antiviral research at the Rega Institute (KU leuven), now 50 years old. PMID- 15535045 TI - Recent advances in the synthesis of the carbocyclic nucleosides as potential antiviral agents. AB - Compared with 4'-oxonucleosides, there have been far fewer systematic structure activity relationship studies on carbocyclic nucleosides as antiviral and antitumour agents. This is mainly because of the synthetic problems in preparing the carbasugars. However, the recent discovery of the ring-closing metathesis (RCM) (a powerful tool for the preparation of 5-membered carbasugar via C-C bond formation) has made it possible to synthesize the key carbasugars to a preparative scale. This review summarizes the asymmetric syntheses of carbasugars and carbocyclic nucleosides, using an RCM reaction as a key step. Furthermore, the review includes valuable information for designing and synthesizing novel carbocyclic nucleosides. PMID- 15535046 TI - Recent clinical experience with famciclovir--a "third generation" nucleoside prodrug. AB - The herpesviruses continue to produce considerable morbidity in man. Once infected with herpes simplex (HSV), the virus remains dormant within the nervous system and may reactivate if provoked by stress, trauma and/or other factors. To date, there is no cure, but antiviral medication can reduce duration and severity of symptoms and prophylaxis can suppress recurrent episodes of disease. The second-generation guanosine nucleosides, acyclovir and penciclovir, are effective inhibitors with low toxicity; both, however, have relatively low oral bioavailability. Subsequently, the orally bioavailable prodrugs valaciclovir and famciclovir have been introduced. These compounds offer high oral bioavailabilty and deliver acyclovir and penciclovir, respectively, to the target cells by means of more convenient dosing schedules. This short review points to recent experience with famciclovir in the management of HSV and varicella-zoster virus. PMID- 15535047 TI - Variability in the PR and RT genes of HIV-1 isolated from recently infected subjects. AB - To study variability in reverse transcriptase (RT) and protease (PR) genes of HIV 1 isolated from recently infected patients identified between 1997 and 2003, sequences were obtained on the RT and PR genes of viruses harvested from plasma of 121 non-treated subjects who had undergone primary HIV infection. The degree of dissimilarity between the viruses studied and a reference HIV-1 subtype B strain (LAV-1) was calculated for each of RT and PR by counting all of the nucleotide substitutions that could be identified. Mutations associated with drug resistance were excluded from analysis. We observed a mean percentage of variation in the RT and PR genes of the viruses analysed of 0.42% between the years 1997--2003 (P<0.01). In PR, the mean variation was 0.71% (P<0.05), while that in RT was 0.3% (P<0.05). Increased diversity was also observed among nucleotides conferring amino-acid changes, although no significant differences in patterns of nucleotide substitutions were apparent over the period of analysis. In conclusion, variability in the RT and PR genes of viruses from recently infected patients has increased over time in a manner that is independent of variability attributable to HIV drug resistance. PMID- 15535048 TI - Treatment of mannan-enhanced influenza B virus infections in mice with oseltamivir, ribavirin and viramidine. AB - Mannan, a polysaccharide preparation from Saccharomyces cerevisiae, has previously been shown to enhance influenza virus replication in mice by inhibiting host defense collectins. The use of mannan in infections may serve to broaden the types of influenza viruses that can be studied in rodent infection models. When mannan was co-administered with influenza B/Sichuan/379/99 virus to mice, the animals died from the infection, whereas mice infected with only virus survived. Three types of influenza A (H1N1) and another influenza B (Hong Kong/330/01) virus infection were also enhanced by mannan, but not four types of influenza A (H3N2) viruses. Mannan was used at 0.16 or 0.5 mg/mouse for optimal disease-enhancing activity using influenza B/Sichuan/379/99 virus. Using this model, influenza B/Sichuan/379/99 infections were treated with oseltamivir, ribavirin or viramidine (the carboxamidine derivative of ribavirin). When oral gavage treatments started 4 h before virus and mannan challenge, oseltamivir was effective at 2.5, 5 and 10 mg/kg/day. Ribavirin was active at 20, 40 and 80 mg/kg/day. Viramidine was effective at 80 and 160 mg/kg/day but not at 40 mg/kg/day. Active drug doses improved lung consolidation scores and lung weights, with decreases in lung virus titres also noted. Arterial oxygen saturation values in treated groups were significantly better than those of the placebo group on days 7-11 of the infection. Oseltamivir (5 mg/kg/day) and ribavirin (40 mg/kg/day) were used alone and in combination to determine how late after infection they could be beneficially administered. Ribavirin alone was very effective (90-100% survival of mice) when treatments started as late as 3 days after infection. Forty percent survival was evident even when treatments started 4 days post-infection. Oseltamivir was active starting treatments 1 day after virus exposure, but lost considerable efficacy when treatments began after that time. The combination of ribavirin and oseltamivir appeared to be no better than ribavirin alone, due to the stronger beneficial effect of ribavirin in this model. The overall results demonstrate that mannan can be used to enhance certain non-lethal influenza virus infections sufficiently to allow antiviral studies. PMID- 15535049 TI - Synthesis, physicochemical and pharmacokinetic studies of potential prodrugs of beta-L-2'-deoxycytidine, a selective and specific anti-HBV agent. AB - beta-L-2'-Deoxycytidine (beta-L-dC) is a potent, selective and specific anti hepatitis B virus (HBV) agent. To improve its oral bioavailability, several derivatives involving sugar or base acylation, as well N4-derivatization with an N,N-(dimethylamino)methylene function, were synthesized. The physicochemical characteristics (including chemical stabilities, solubilities and distribution coefficient values) and pharmacokinetics of these compounds were determined and compared with those of the parent drug, beta-L-dC. PMID- 15535050 TI - Therapeutic basis of vidarabine on adenovirus-induced haemorrhagic cystitis. AB - When adenovirus causes haemorrhagic cystitis in immunocompromised patients, vidarabine is used for its treatment because therapeutic choice is limited. Although vidarabine has been reported to be effective for these patients, its therapeutic basis has not yet been established. Vidarabine dose-dependently inhibited viral replication as assessed by a yield reduction assay. Viral protein synthesis was dose-dependently inhibited by vidarabine but not at all by acyclovir, and the degree of inhibition by vidarabine was different for each of the viral proteins, ranging from 0-40% of the untreated control. These results indicated the specificity and mechanism of action of vidarabine against adenovirus. The concentration of vidarabine and its metabolite in the bladder is suggested to exhibit effective anti-adenoviral activity in suppressing the replication of adenovirus. Thus, our results support vidarabine therapy as a possible candidate for adenovirus-induced haemorrhagic cystitis in immunocompromised patients. PMID- 15535051 TI - A model for mechanotransduction in cardiac muscle: effects of extracellular matrix deformation on autocrine signaling. AB - We present a computational model and analysis of the dynamic behavior of epidermal growth factor receptor (EGFR) signaling in cardiac muscle tissue, with the aim of exploring transduction of mechanical loading into cellular signaling that could lead to cardiac hypertrophy. For this purpose, we integrated recently introduced models for ligand dynamics within compliant intercellular spaces and for the spatial dynamics of intracellular signaling with a positive feedback autocrine circuit. These kinetic models are here considered in the setting of a tissue consisting of cardiomyocytes and blood capillaries as a structural model for the myocardium. We show that autocrine EGFR signaling can be induced directly by mechanical deformation of the tissue and demonstrate the possibility of self organization of signaling that is anisotropic on the tissue level and can reflect anisotropy of the mechanical deformation. These predictions point to the potential capabilities of the EGFR autocrine signaling circuit in mechanotransduction and suggest a new perspective on the cardiac hypertrophic response. PMID- 15535052 TI - Estimation of distance between a unipolar recording electrode and a myocardial bundle based on signal characteristics. AB - The aim of the present paper, is the estimation of the distance between an electrode used as a recording site of the extracellular potential field and a surviving myocardial bundle. The importance of the reliable solution of this problem lies among others in controlling ablation. For our purposes one dimensional propagation is considered and current sources are activated along a cable simulating the propagating waves with constant velocity. Different models of current sources are explored. By use of these models, the corresponding functions expressing extracellular potentials are calculated, using the volume conductor equation. This way, extracellular potentials are modeled as parametric functions of longitudinal distance, while perpendicular distance, current source strength, and other factors related to the propagated wave are parameters of the functions. Simulated annealing is applied for model parameter estimation and appropriate Time Domain and Wavelet Domain cost functions are investigated. Different combinations of model and cost function are evaluated regarding the accuracy of distance estimation. A continuous source model function with a wavelet cost function was found to be the most accurate combination. The accuracy of distance estimation is related to the selected source model and to the actual distance of recording in a nonmonotonic way. PMID- 15535053 TI - Assessment of chaotic parameters in nonstationary electrocardiograms by use of empirical mode decomposition. AB - This study addressed the issue of assessing chaotic parameters from nonstationary electrocardiogram (ECG) signals. The empirical mode decomposition (EMD) was proposed as a method to extract intrinsic mode functions (IMFs) from ECG signals. Chaos analysis methods were then applied to the stationary IMFs without violating the underlying assumption of stationarity. Eight ECG data sets representing normal and various abnormal rhythms were obtained from the American Heart Associate Ventricular Arrhythmia database. The chaotic parameters including Lyapunov exponent, entropy, and correlation dimension were computed. The results consistently showed that the 10th IMF (IMF-10) was stationary and preserved sufficient nonlinearity of the ECG signals. Each IMF-10 from the data sets (n = 8) gave a positive dominate Lyapunov exponent (0.29-0.64, p < 0.0001), a positive entropy (0.039-0.061, p < 0.0001), and a noninteger correlation dimension (1.1 1.9). These were evidences of a chaotic dynamic system. We therefore concluded that the original ECG signals must also have chaotic properties. The chaotic parameters did not show significant differences among the eight data sets representing normal sinus rhythm and various abnormalities. This study has demonstrated an effective way to characterize nonlinearities in nonstationary ECG signals by combining the empirical mode decomposition and the chaos analysis methods. PMID- 15535054 TI - Elastic behavior of porcine coronary artery tissue under uniaxial and equibiaxial tension. AB - The aim of this study was to characterize the nonlinear anisotropic elastic behavior of healthy porcine coronary arteries under uniaxial and equibiaxial tension. Porcine coronary tissue was chosen for its availability and similarity to human arterial tissue. A biaxial test device previously used to test human femoral arterial tissue samples (Prendergast, P. J., C. Lally, S. Daly, A. J. Reid, T. C. Lee, D. Quinn, and F. Dolan. ASME J. Biomech. Eng., Vol. 125, pp. 692 699, 2003) was further developed to test porcine coronary tissue specimens. The device applies an equal force to the four sides of a square specimen and therefore creates a biaxial stretch that demonstrates the anisotropy of arterial tissue. The nonlinear elastic behavior was marked in both uniaxial and biaxial tests. The tissue demonstrated higher stiffness in the circumferential direction in four out of eight cases subjected to biaxial tension. Even though anisotropy is demonstrated it is proposed that an isotropic hyperelastic model may adequately represent the properties of an artery, provided that an axial stretch is applied to the vessel to simulate the in vivo longitudinal tethering on the vessel. Isotropic hyperelastic models based on the Mooney-Rivlin constitutive equation were derived from the test data by averaging the longitudinal and circumferential equibiaxial data. Three different hyperelastic models were established to represent the test specimens that exhibited a high stiffness, an average stiffness, and a low stiffness response; these three models allow the analyst to account for the variability in the arterial tissue mechanical properties. These models, which take account of the nonlinear elastic behavior of coronary tissue, may be implemented in finite element models and used to carry out preclinical tests of intravascular devices. The errors associated with the hyperelastic models when fitting to both the uniaxial and equibiaxial data for the low stiffness, average stiffness, and high stiffness models were found to be 0.836, 5.206, and 2.980, respectively. PMID- 15535055 TI - Adaptive predictive control of arterial blood pressure based on a neural network during acute hypotension. AB - In acute hypotension, an automated drug infusion system to control mean arterial blood pressure (MAP) has not been previously studied, though many investigations have examined the use of vasodilating drugs to control MAP in postoperative hypertension. Therefore, we examined an automated control of MAP during acute hypotension using a neural network (NN) approach. A proportional-integral derivative (PID) control, an adaptive predictive control using a NN (APC(NN)), a combined control of APC(NN) and PID (APC(NN-PID)), a fuzzy control, and a model predictive control were tested in computer simulation based on the MAP response to norepinephrine (NE) of 25 microg ml(-1). In six anesthetized rabbits, using the NE of 25 microg ml(-1), the PID control, APC(NN), and APC(NN-PID) prevented severe hypotension compared to an uncontrolled condition. Under PID control, four of the six animals showed MAP oscillation. Using NE of 50 microg ml(-1), the rabbits recovered from acute hypotension for all systems tested but showed sustained MAP oscillation during PID control. In conclusion, utilization of a NN for adaptive predictive control systems could facilitate the development of an automated drug infusion apparatus because it provides robust control even when acute or large perturbations and inter-individual differences in the sensitivity to therapeutic agents occur. PMID- 15535056 TI - In situ thermal denaturation of proteins in dunning AT-1 prostate cancer cells: implication for hyperthermic cell injury. AB - The in situ thermal protein denaturation and its correlation with direct hyperthermic cell injury in Dunning AT-1 prostate tumor cells were investigated in this study. The in situ thermal protein denaturation was studied using both Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). The FTIR spectra at different temperatures show changes in protein secondary structure (from alpha helix to extended beta sheet) during in situ thermal protein denaturation within AT-1 cells. Calorimetric studies using DSC show that endothermic heat release is associated with the in situ thermal protein denaturation. Furthermore, both the secondary structure changes detected by FTIR and the calorimetric changes detected by DSC were quantified and the kinetics of the overall in situ thermal protein denaturation was derived under different heating conditions. The onset temperature where the overall in situ thermal protein denaturation is first detectable was found to be scanning rate dependent (approximately 41 degrees C at 2 degrees C min(-1) and approximately 44 degrees C at 5 degrees C min(-1)). The kinetics of the overall in situ thermal protein denaturation was derived from both DSC and FTIR measurements and was fit using kinetic and statistical models. The kinetic data determined by FTIR and DSC under the same heating conditions match well with each other. The activation energy of the overall in situ thermal protein denaturation is found to be strongly dependent on the temperature range considered (the activation energy ranges from approximately 110 kJ mol(-1) between 44 and 90 degrees C to approximately 750 kJ mol(-1) between 44 and 50 degrees C). However, its dependence on heating rate is negligible. Several denaturation peaks, including a dominant one between approximately 62 and 65 degrees C, are identifiable from both the DSC and the FTIR results. To investigate directly the relationship between thermally induced cell injury and the in situ thermal protein denaturation, both acute (propidium iodide dye exclusion, assessed 3-h postthermal treatment) and chronic (clonogenics, assessed 7-day postthermal treatment) cell injury were quantified using AT-1 cells prepared under the same conditions as for the DSC protein studies. Comparisons of the results from the cell injury studies and the DSC protein denaturation studies show that the overall in situ thermal protein denaturation correlates well with both the acute and the chronic cell injury, which suggests that overall in situ thermal protein denaturation is an important mechanism of direct hyperthermic cell injury in AT-1 cells at the macromolecular level. PMID- 15535057 TI - Multi-modal medical image integration to optimize radiotherapy planning in lung cancer treatment. AB - This work presents a method for CT and PET image registration, and multi-modal analysis, to optimize radiotherapy planning in lung cancer treatment. The method relies on an image registration technique based on fiducial external markers to realign, spatially, PET images with the CT spatial reference system. The method was set up for clinical use in radiotherapy, allowing minimal modifications to be introduced in the management of patients undergoing radiation treatment. The accuracy of the registration technique was evaluated on patient studies in terms of Target Registration Error and was found to be less than 6.40 mm. The method was applied in the treatment planning of five patients affected by non-small-cell lung cancer, revealing the usefulness of PET/CT integration in delineating the extension of both the tumor mass and the tissues involved in the neoplastic process. Moreover, the functional information provided by PET often led to alterations in the treatment planning, changing the size and/or direction of radiation portals. The proposed method for PET/CT integration has been confirmed as being useful for optimizing radiotherapy planning in lung cancer treatment. PMID- 15535058 TI - Changes in the biaxial viscoelastic response of the urinary bladder following spinal cord injury. AB - In order to gain a deeper understanding of bladder function, it is necessary to study the time-dependent response of the bladder wall. The present study evaluated and compared the viscoelastic behaviors of normal and spinal cord injured (SCI) rat bladder wall tissue using an established rat model and planar biaxial stress relaxation tests. Bladders from normal and spinalized (3 weeks) rats were subjected to biaxial stress (either 25 or 100 kPa in each loading direction) rapidly (in 50 ms) and subsequently allowed to relax at the constant stretch levels in modified Kreb's solution (in the absence of calcium; with no smooth muscle tone) for 10,000 s. We observed slower and therefore less stress relaxation in the SCI group compared to the normal group, which varied with the stress-level. These experimental results were fitted (r2 > 0.98) to a reduced relaxation function. Furthermore, biochemical assays revealed that the collagen content of SCI rat bladders was significantly (p < 0.05) lower by 43%, while the elastin content was significantly (p < 0.001) higher by 260% than that of normal bladders. These results suggest that SCI and the associated urologic functional changes induce profound tissue remodeling, which, in turn, provided the structural basis for the alterations in the complex, time-dependent mechanical behavior of the urinary bladder wall observed in the present study. PMID- 15535059 TI - Thoracic gas volume measurements in paralyzed mice. AB - We have previously measured thoracic gas volume (VTG) in spontaneously breathing mice using a whole body plethysmograph and have now extended our technique to allow for V(TG) measurements during paralysis. BALB/c mice were anesthetized and placed in a body-box and ventilated via a tracheostomy cannula through the box wall. Box pressure (Pb) and tracheal pressure (Pao) were measured during spontaneous breathing, and again after paralysis while mechanically compressing the chest. V(TG) was much larger after paralysis (0.49+/-0.06 ml, positive end expiratory pressure = 2 cmH2O) when compared with spontaneous breathing (0.31+/ 0.01 ml). External chest compression produced looping in the plots of Pb versus Pao that was attributable to gradual changes in Pb upon release of the mechanical chest compression and had the character of thermal transients. Under the assumption that the rate of heating of the air in the chamber was proportional to the pressure applied to the animal's chest, and that any increase in air temperature was dissipated by heat absorption by the chamber walls, we developed an algorithm that corrected for the thermal events. This yielded similar results for V(TG) (0.30+/-0.02 ml) as obtained during spontaneous efforts. Our method may prove particularly useful when paralysis is required for the precise measurement of lung mechanics. PMID- 15535060 TI - Effects of tetraethylammonium on Kx channels and simulated light response in rod photoreceptors. AB - Rod photoreceptors express a unique type of noninactivating potassium channels, Kx channels, which play an important role in setting the dark potential and participate in shaping the light response. Biophysical studies of Kx channels are limited. For example, the effects of a conventional blocker of potassium channels, tetraethylammonium (TEA), on Kx channels have not been extensively studied. Here we demonstrate that TEA blocks Kx channels, one molecule of TEA being sufficient to block the channel. Half of the Kx current was inhibited at K0.5 = 5.6 mM. The TEA-induced block of Kx channels depended on extracellular potassium: the higher the potassium concentration, the stronger the block. Using TEA, we blocked potassium channels to reveal their role in shaping the simulated light response (SLR) of rods. We showed that TEA slowed down SLR and sometimes caused generation of action potentials. We developed a complete computer model of the rod, which accurately reproduced the main features of the light response and allowed us to demonstrate that it was suppression of Kx channels that was essential for slowing SLR and increasing excitability of rods. The results reported in this work further establish the importance of Kx channels in rod photoreceptor function. PMID- 15535061 TI - Anisotropic, three-dimensional deformation of single attached cells under compression. AB - Quantifying three-dimensional deformation of cells under mechanical load is relevant when studying cell deformation in relation to cellular functioning. Because most cells are anchorage dependent for normal functioning, it is desired to study cells in their attached configuration. This study reports new three dimensional morphometric measurements of cell deformation during stepwise compression experiments with a recently developed cell loading device. The device allows global, unconfined compression of individual, attached cells under optimal environmental conditions. Three-dimensional images of fluorescently stained myoblasts were recorded with confocal microscopy and analyzed with image restoration and three-dimensional image reconstruction software to quantify cell deformation. In response to compression, cell width, cross-sectional area, and surface area increased significantly with applied strain, whereas cell volume remained constant. Interestingly, the cell and the nucleus deformed perpendicular to the direction of actin filaments present along the long axis of the cell. This strongly suggests that this anisotropic deformation can be attributed to the preferred orientation of actin filaments. A shape factor was introduced to quantify the global shape of attached cells. The increase of this factor during compression reflected the anisotropic deformation of the cell. PMID- 15535063 TI - The phylogeny of Porinaceae (Ostropomycetidae) suggests a neotenic origin of perithecia in Lecanoromycetes. AB - The family Porinaceae (Trichotheliales) is characterized by perithecial ascomata of ascohymenial origin. The phylogenetic position of this family in the system of ascomycetes has been uncertain and is investigated using mtSSU rDNA sequences. The dataset consists of lichenized representatives of major ascomycete lineages, including those that were previously suspected as relatives of Porinaceae, such as Pyrenulaceae. The dataset was subjected to a Bayesian phylogenetic analysis implementing a Metropolis Coupled Markov Chain Monte Carlo method. The analysis confirms previous classification of the apothecial Gomphillaceae near to Graphidaceae, and suggests that the pyrenocarpous Porinaceae are also close to Graphidaceae, Gyalectaceae, and Stictidaceae. The subclass Ostropomycetidae is here suggested to include the families Gomphillaceae, Gyalectaceae, Graphidaceae (incl. Thelotremataceae), Porinaceae, and Stictidaceae. A special type of hemiangiocarpous ontogeny of the ascomata is shared throughout the Ostropomycetidae, and the closed fruit bodies of Porinaceae are apparently a result of a neotenic ontogeny. This is associated with special hymenial characters: rather thin-walled narrow asci, and a different consistency of the hymenial gels. PMID- 15535062 TI - A rapid, reliable method to isolate high quality endothelial RNA from small spatially-defined locations. AB - Phenotypic heterogeneity of the endothelium has important implications for cell sourcing for cardiovascular tissue engineered devices and is fundamental to many cardiovascular diseases. A critical first step to identifying genetic regulators associated with particular endothelial phenotypes is reliable isolation of pure RNA from the cell subpopulations of interest. We present here a rapid method for the isolation of endothelial RNA from small spatially-defined locations, illustrated for two sides of the porcine aortic valve. Endothelial cells were retrieved from fresh tissue by freezing them to a glass substrate, from which they were lysed in guanidine thiocyanate buffer for RNA isolation. Valve endothelial cells isolated by this technique stained positively for CD31 and von Willebrand factor, consistent with an endothelial phenotype, with no evidence of contamination by alpha-smooth muscle actin-positive valve interstitial cells or CD45-positive leukocytes. RNA integrity was excellent in 80% of the samples, with over 100 ng of total RNA typically obtained from each side of the valve. This rapid method yields high quality pure endothelial RNA in sufficient quantities for amplification and subsequent use in device-, cell-, and location-specific transcriptional profiling by microarray technologies, and thus facilitates studies of spatial gene regulation. PMID- 15535064 TI - A synopsis and re-circumscription of Neurospora (syn. Gelasinospora) based on ultrastructural and 28S rDNA sequence data. AB - Neurospora and Gelasinospora are traditionally distinguished by the ornamentation pattern of the surface of their ascospores, which are ribbed in the former and pitted in the latter. However, a detailed examination of the morphology of numerous strains of most of the species of both genera confirm the hypothesis that there are not enough criteria to distinguish them from each other. The names Neurospora and Gelasinospora are synonymized and the circumscription of the genus Neurospora amended. Partial sequences of the 28S rDNA gene from 27 species of both genera were analysed to infer their phylogenetic relationships. Species of the two genera were interspersed in the different clades and confirmed that they are genetically very similar. The grouping obtained demonstrates that the morphology of the episporial-layer of the ascospores is an informative phylogenetic character. Two recent isolates from soils of Nigeria and Spain, which could not be classified as any known species of Neurospora are described, illustrated, and recognized as new: N. nigeriensis and N. uniporata spp. nov. A synopsis and key to the 49 species of Neurospora now recognized in the genus is presented, and the new genus Pseudogelasinospora described to accommodate P. amorphoporcata (syn. Gelasinospora amorphoporcata comb. nov.). PMID- 15535065 TI - Two novel kickxellalean fungi, Mycoemilia scoparia gen. sp. nov. and Ramicandelaber brevisporus sp. nov. AB - Mycoemilia scoparia gen. sp. nov. is described as a new member of Kickxellales. It is characterized by lageniform sporocladia produced acrogenously in mass and bears wet and fusiform spores on the sporocladia. Ramicandelaber brevisporus sp. nov. is distinguished from the type species of the genus, R. longisporus, by producing much shorter asexual spores, (3-)8(-13) long fertile branches arising from a globose body, and lateral branches. PMID- 15535066 TI - Variations in rDNA ITS of Czech Armillaria species determined by PCR and HPLC. AB - We analysed 40 isolates of species Armillaria. borealis, A. cepistipes, A. gallica, A. mellea, A. ostoyae and A. tabescens, mostly collected in the Czech Republic, by PCR-RFLP of the ITS rRNA genes using the restriction endonucleases AluI, HinfI and MboI. Restriction fragments were analysed by ion-exchange high performance liquid chromatography which proved to be more useful informative, and less time-consuming than classical electrophoresis on agarose gel. The HPLC method enabled detection of some heterozygous strains. HinfI discriminated between all six species. Ten isolates were sequenced to confirm changes in restriction sites found by restriction analysis. Cluster analysis based on the restrictions patterns of restriction endonucleases AluI and HinfI divided the analysed species into three groups. The first and the most distant group contained all A. mellea isolates, the second group was formed by A. tabescens and the third group contained species A. borealis, A. cepistipes, A. gallica and A. ostoyae. The A. tabescens group was very homogenous regardless of the origin of isolates (Czech Republic, France and Finland). PMID- 15535067 TI - Uniting Tricholoma sulphureum and T. bufonium. AB - The taxonomic status and relationship of Tricholoma sulphureum and the similar T. bufonium were investigated using different sets of characters. These included morphological data on fruit bodies, ecological and chorological data, and analysis of the sequence data obtained for the ITS of basidiomes of different ecological and geographic origin. Moreover, the ectomycorrhizas formed by T. bufonium on Abies alba and Quercus sp. were characterised, and anatomical features compared with those of T. sulphureum mycorrhizas on coniferous and broad leaved host trees. Our results revealed extensive ITS variation in members of the T. sulphureum group, but this variation was not correlated with morphology, ecology, or geographical distribution. We conclude that T. bufonium cannot be maintained as an autonomous taxon and should be treated as an infraspecific variant of T. sulphureum. PMID- 15535068 TI - Phytophthora alni sp. nov. and its variants: designation of emerging heteroploid hybrid pathogens spreading on Alnus trees. AB - In 1993 a destructive new Phytophthora pathogen of riparian Alnus trees was discovered in the UK and subsequently shown to be present in other parts of Europe. The new Phytophthora comprised a group of emergent heteroploid hybrids, probably between P. cambivora and a species related to P. fragariae. These included a common, near tetraploid standard hybrid, the presumptive allopolyploid; and four scarcer major variant types with chromosome numbers intermediate between diploid and tetraploid, named the Swedish, Dutch, German and UK variants. The standard hybrid type is formally designated here as Phytophthora alni subsp. alni. The Swedish variant is designated as P. alni subsp. uniformis; and the Dutch, German and UK variants collectively as P. alni subsp. multiformis. The properties of the Dutch, German and UK variants within subsp. multiformis are informally described. The problems of designating emergent species hybrids under the International Code of Botanical Nomenclature and the reasons for the taxonomic choices made are discussed. PMID- 15535069 TI - Streptopodium caricae sp. nov., with a discussion of powdery mildews on papaya, and emended descriptions of the genus Streptopodium and Oidium caricae. AB - A new powdery mildew infecting papaya (Carica papaya) in Brazil, Streptopodium caricae sp. nov., is described. The species is compared with other anamorphic Erysiphales known to infect papaya: Oidiopsis sicula, Ovulariopsis papayae, Oidium caricae, O. papayae, O. caricicola, O. indicum, O. caricae-papayae, Podosphaera (syn. Sphaerotheca) spp., and Erysiphe spp. An emended description Streptopodium and a key to the anamorphs of powdery mildews on papaya are also presented. A re-examination of the type material of Phyllactinia caricaefolia showed that conidia in this material are dimorphic, indicating that its anamorph does not belong to Ovulariopsis and that the teleomorph is not conspecific with Phyllactinia guttata. Oidium caricae, the common powdery mildew of papaya, was re examined, recognized as a member of subgenus Pseudoidium, an emended description was prepared, and a new type was indicated. O. papayae was recognized as a synonym of O. caricae, and many of the records of this fungus are considered to be doubtful or incorrect, either omitting a description of the fungus or including a description or illustration of an euodium conidiophore morphology. PMID- 15535070 TI - Identification and characterization of Eutypa leptoplaca, a new pathogen of grapevine in Northern California. AB - Although Eutypa lata is the main agent of Eutypa dieback of grapevine, another species of Eutypa has been isolated from diseased grapevines in Northern California. Stromata of this recently discovered Eutypa were also collected from Acer macrophyllum, A. negundo, and Fraxinus latifolia in the vicinity of vineyards, and appeared commonly on Umbellularia californica in some mixed evergreen forests of Napa and Sonoma counties. This second species of Eutypa was distinguished from E. lata because of the sulcate ostiole of the perithecium and smaller ascospores. A morphological comparison with type specimens revealed identical features between the Californian isolates and E. leptoplaca sensu Rappaz (1987). This identification was confirmed through phylogenetic analyses of Eutypa spp. based on the complete sequence of the internal transcribed spacer (ITS) of the rDNA and partial sequence of the beta-tubulin gene. These analyses also separated collections of E. maura, E. sparsa, E. lejoplaca, E. tetragona, E. leptoplaca and E. lata, confirming the previously proposed species concepts. The pathogenicity of E. leptoplaca on grapevine was established using isolates collected from Vitis vinifera, U. californica, and A. macrophyllum. The importance of E. leptoplaca in relation to Eutypa dieback and its role as a necrotrophic pathogen are discussed. PMID- 15535071 TI - Low genetic diversity among isolates of the nematode-trapping fungus Duddingtonia flagrans: evidence for recent worldwide dispersion from a single common ancestor. AB - The genetic variation of Duddingtonia flagrans, which has become a promising biocontrol agent of animal parasitic nematodes, was investigated in a worldwide collection of 22 isolates. We analysed the sequence variation in four nuclear genes, tubA (beta-tubulin), CMD1 (calmodulin), EF1alpha (translation elongation factor 1alpha), and PII (extracellular serine protease). 1428 aligned base pairs (bp) were analysed from the four genes, including 709 bp of introns. In addition, the variations in three anonymous genomic regions comprising 1155 bp were examined. Three single nucleotide polymorphisms (SNPs) were detected in the seven loci, none of them in the protein encoding genes. The genetic variation was significantly higher in the nematode-trapping fungus Arthrobotrys oligospora, the closest evolutionary relative to D. flagrans. Analysis of 12 isolates of A. oligospora revealed 30 SNPs in tubA, CMD1, EF1alpha and PII. The genetic variation in the isolates of D. flagrans was further examined using AFLP analysis. Five primer combinations were used to detect 159 bands, of which 94 (59.1%) were polymorphic. A neighbour-joining tree based on the AFLP data showed no clear association between genotype and geographical origin. Furthermore, the AFLP data suggest that D. flagrans is mainly clonal and no recombination could be detected, not even within the same country. The low genetic variation in D. flagrans suggests that this fungus has recently diverged from a single progenitor. Based on estimations of mutation rates, it was calculated that this most recent common ancestor lived about 16,000-23,000 years ago. PMID- 15535072 TI - Methods for the isolation, culture and assessment of the status of anaerobic rumen chytrids in both in vitro and in vivo systems. AB - Anaerobic fungi were isolated from both the rumen and faeces of nine sheep and a cow. A reliable and simple method for the isolation of anaerobic fungi using 24 h rumen incubated milled straw as the inoculum source was developed. We also evaluate the use of chitin measurements as an assay of rumen fungal biomass. Chitin levels were determined from various sample sources (milled barley straw used as the fungal culture substrate in vitro; plant particulate digests from the rumen (PLP) and centrifuged strained rumen fluid (CSRF) using both HPLC and colorimetric methods. Both methods were highly correlated and consequently the simpler colorimetric method was adopted for subsequent studies. There was also a high degree of correlation between anaerobic fungal cellulase activities with the assayed chitin content of milled barley straw cultures over 12 d of an in vitro experiment. The colorimetric chitin assay protocol was then used to assess the diurnal variation and abundance of rumen fungi in in vivo assays. We assessed the distribution of chitin (mg g(-1) dry matter) in various fractions of the strained rumen fluid (SRF) and PLP samples from the rumen of sheep. Chitin was detected in all fractions of strained rumen fluid but the main source of chitin in the samples may be attributed to the fungal biomass. We did not detect any significant differences in chitin levels over a 24 h sampling period. Finally, an SEM study on subsamples of milled straw and plant particulate matter used in the chitin assays, revealed that the pattern of the fungal development on substrate material differs from the culture medium to the rumen. PMID- 15535073 TI - Distribution and estimation of anaerobic zoosporic fungi along the digestive tracts of sheep. AB - The status of anaerobic zoosporic (Chytridiomycota) fungi along the entire digestive tract of sheep was assessed both analytically and microscopically. Digest samples were taken from different segments of the digestive tracts of three newly killed sheep that previously had been used in experimental dietary studies. These digest samples were tested for the presence of rumen fungi by assessing the recovery of live fungi from the samples, direct observation of digested plant remains under the scanning electron microscope (SEM), and using a chitin assay as an estimation of fungal biomass. Live anaerobic fungi were recovered from the abomasum, small and large intestine, caecum and faeces of sheep, but not from the digest samples of rumen and omasum. However, SEM examination of the samples confirmed the presence of fungal structures from all of these organs. In the large intestine and caecum samples the observed sporangial structures were rounded and showed conspicuous surface pitting. Results of the chitin assay indicated that the anaerobic fungi might account for up to 20% of the total microbial biomass in the rumen of sheep. The results of this study support the view that anaerobic fungi may be present as a resistant stage in the lower reaches of the digestive tract. PMID- 15535074 TI - Characteristics and risk factors associated with high-cost Medicaid recipients. AB - Characteristics and risk factors of fee-for-service Medicaid patients (age < 65 yr) with high health care costs were assessed by analyzing Ohio's Medicaid claims database. High-cost recipients were defined as those with average monthly Medicaid expenses at or above the 90th percentile. The records of 10,582 high cost patients and 11,045 comparison patients were examined for clinical comorbidity, mortality status, enrollment, and demographic factors, using logistic and logarithmic multiple regression. Researchers found that a Medicaid recipient had the greatest odds of being in the following groups: dying (odds ratio = 4.0), disabled (2.2), urban resident (1.8), and male (1.3). They concluded that state and federal efforts to control Medicaid expenditures should focus on those high-costrecipients and examine their service utilization. PMID- 15535075 TI - Better health care through better competitive incentives. PMID- 15535076 TI - Improving disease management in the United Kingdom: what can be learned from U.S. experience? AB - Chronic diseases are a large and growing burden for health services. The British National Health Service is set up in a way that has many advantages in the management of chronic diseases, but lessons from U.S. managed care plans show that several changes could be implemented to improve the efficiency and effectiveness of care for patients with chronic conditions. These include the introduction of limited market competition; financial incentives, particularly across primary and secondary care; increased patient self-help; and improved clinician-manager relationships. PMID- 15535077 TI - Comparing specialists' performance to improve quality. AB - The health care community must address criticisms that the quality of health care in the United States is inadequate. This article describes one successful program in a large, not-for-profit health plan in which key aspects of physician specialty groups' performance are evaluated by both patients and the generalists who refer them. The results of the evaluations, which include comparisons among practices, are provided to the specialist groups, who use the information to improve their quality of care. The authors also discuss features that are essential to the success of such quality-improvement programs. PMID- 15535078 TI - Designating drug classes: model guidelines for Medicare. PMID- 15535079 TI - The role of the Employee Retirement Income Security Act of 1974. AB - On June 21, 2004, the Supreme Court ruled on Aetna Health Inc. v. Davila, the latest in a long line of decisions regarding the scope of Employee Retirement Income Security Act of 1974 (ERISA) preemption and the remedies available to participants of ERISA plans. After taking what many observers perceived to be a detour over the last few years, the Davila decision returned the Court to a more expansive interpretation of ERISA's preemption clause. PMID- 15535080 TI - The cost effectiveness of universal diabetes screening. PMID- 15535081 TI - Arctic rabies--a review. AB - Rabies seems to persist throughout most arctic regions, and the northern parts of Norway, Sweden and Finland, is the only part of the Arctic where rabies has not been diagnosed in recent time. The arctic fox is the main host, and the same arctic virus variant seems to infect the arctic fox throughout the range of this species. The epidemiology of rabies seems to have certain common characteristics in arctic regions, but main questions such as the maintenance and spread of the disease remains largely unknown. The virus has spread and initiated new epidemics also in other species such as the red fox and the racoon dog. Large land areas and cold climate complicate the control of the disease, but experimental oral vaccination of arctic foxes has been successful. This article summarises the current knowledge and the typical characteristics of arctic rabies including its distribution and epidemiology. PMID- 15535082 TI - Sperm morphology in Estonian and Tori breed stallions. AB - The standard procedure for assessing the breeding potential of a stallion includes the parameter total number of spermatozoa classified as morphologically normal. This study investigated sperm morphology of fresh semen in randomly chosen Estonian (E, n = 8) and Tori (T, n = 7) breed stallions with proven fertility. Two ejaculates were examined from each stallion. An aliquot from each ejaculate was fixed in 1 mL formol-saline immediately after collection and examined with phase-contrast microscope at a magnification 1000x for all types of morphological abnormalities. Furthermore smears were prepared and stained according to Williams (carbolfuchsin-eosin) for a more detailed examination of the sperm heads with light microscope at a magnification 1000x. Analysis of variance was applied to the data, and results are presented as LSmeans (+/- SE). One T stallion that had a disturbance in the spermatogenesis and one 22-year-old E stallion were not included in the analyses. The T stallions had on average 57.5 +/- 4.1% and the E-stallions 74.4 +/- 3.8% morphologically normal spermatozoa (p = 0.012). In 4 of 7 T stallions and 7 of 8 E stallions both ejaculates had > 50% morphologically normal spermatozoa. There was a significant difference between breeds in mean percentage of proximal droplets (17.3 +/- 2.7% and 2.9 +/- 2.5% for T and E stallions, respectively; p = 0.003). PMID- 15535083 TI - Bacteriological investigation of infectious keratoconjunctivitis in Norwegian sheep. AB - Contagious keratoconjunctivitis is a rather common disease in Norwegian sheep. Since the knowledge of its aetiology is limited, the present study was performed to determine the microorganisms involved. Local veterinarians throughout the country collected conjunctival swabs from both sick (n = 43) and healthy (n = 42) sheep on 15 farms with outbreaks of ovine keratoconjunctivitis, and further from healthy sheep (n = 50) on 17 farms not showing any signs of conjunctival disease. All samples were cultivated for bacteria and mycoplasma. Listeria monocytogenes was isolated from 3 cases (1%) in one single herd. Staphylococcus aureus (5%), Corynebacterium spp. (2%) and Escherichia coli (4%) were isolated only in herds with keratoconjunctivitis, but from both sick and healthy animals. Moraxella (Branhamella) ovis was isolated from 28% of sampled animals in affected herds and from 10% of sampled animals in healthy herds. The corresponding numbers for Moraxella spp. were 9%/12%, for Pseudomonas spp. 7%/8%, for Staphylococcus spp. 22//22%, for Bacillus spp. 12%/14%, for Micrococcus spp. 6%/2% and for Streptococcus/Enterococcus spp. 2%/2%. Mycoplasma conjunctivae was isolated from 16 animals with keratoconjunctivitis (37%) and from 3 animals without clinical signs (7%) in farms with keratoconjunctivitis. In farms without clinical signs of keratoconjunctivitis, M. conjunctivae was isolated in 4 animals (8%). To our knowledge, this is the first time M. conjunctivae has been isolated in Norway. Other predisposing agents found were Moraxella (Branhamella) ovis and Listeria monocytogenes. The etiological importance of different microorganisms in ovine keratoconjunctivitis seems to vary; some are probably only present as secondary invaders. Other possible causes of ovine keratoconjunctivitis in Norway, such as Chlamydia psittaci, remain to be investigated. PMID- 15535084 TI - Studies on calf diarrhoea in Mozambique: prevalence of bacterial pathogens. AB - The prevalence of diarrhoea in calves was investigated in 8 dairy farms in Mozambique at 4 occasions during 2 consecutive years. A total of 1241 calves up to 6 months of age were reared in the farms, and 63 (5%) of them had signs of diarrhoea. Two farms had an overall higher prevalence (13% and 21%) of diarrhoea. Faecal samples were collected from all diarrhoeal calves (n = 63) and from 330 healthy calves and analysed for Salmonella species, Campylobacter jejuni and enterotoxigenic Escherichia coli (ETEC). Salmonella spp. was isolated in only 2% of all calves. Campylobacter was isolated in 11% of all calves, irrespective of health condition, and was more frequent (25%) in one of the 2 diarrhoeal farms (p = 0.001). 80% of the isolates were identified as C. jejuni. No ETEC strains were detected among the 55 tested strains from diarrhoeal calves, but 22/55 (40%) strains from diarrhoeal calves and 14/88 (16%) strains from healthy calves carried the K99 adhesin (p = 0.001). 6,757 E. coli isolates were typed with a biochemical fingerprinting method (the PhenePlate) giving the same E. coli diversity in healthy and diarrhoeal calves. Thus it was concluded: i) the overall prevalence of diarrhoea was low, but 2 farms had a higher prevalence that could be due to an outbreak situation, ii) Salmonella did not seem to be associated with diarrhoea, iii) Campylobacter jejuni was common at one of the 2 diarrhoeal farms and iv) ETEC strains were not found, but K99 antigen was more prevalent in E. coli strains from diarrhoeal calves than from healthy, as well as more prevalent in one diarrhoeal farm. PMID- 15535085 TI - Antimicrobial resistance in Staphylococcus spp., Escherichia coli and Enterococcus spp. in dogs given antibiotics for chronic dermatological disorders, compared with non-treated control dogs. AB - The aim of this study was to evaluate antimicrobial resistance in canine staphylococci, Escherichia coli and enterococci, which were isolated from 22 dogs with pyoderma and a history of previous antibiotic treatment, compared to bacterial isolates from 56 non-treated control dogs. Two isolates of each bacterial species per dog were investigated, if detected. Staphylococcal isolates from dogs with pyoderma (35 isolates) were more resistant to sulphatrimethoprim than the isolates from controls (56 isolates) (57% vs. 25%, p < 0.004). Multiresistance in staphylococci was also more common in dogs with pyoderma (29% vs. 9%, p = 0.02). A similar trend among isolates of E. coli was detected (24 and 74 isolates from treated and control dogs, respectively), but the differences were not significant. Resistance for macrolide-lincosamides was approximately 20% among staphylococci in both groups. Resistance to ampicillin among enterococci was 4%-7%. The age of the dogs might have an impact on resistance: multiresistance among staphylococcal isolates from younger dogs (< or = 5 years) was more common than in older dogs (26 years) (24%, vs. 0%, 63 and 27 isolates, respectively, p = 0.02). Staphylococci in younger dogs were more resistant to tetracycline (48% vs. 11%, p < 0.001) and sulphatrimethoprim (48% vs. 15%, p < 0.01) than those in older dogs. In contrast, the isolates of E. coli from older dogs tended to be more resistant, although a significant difference was detected only in resistance to tetracycline (13% vs. 2% of 40 and 50 isolates respecthely, p = 0.04)). The results of this small study indicate that resistance in canine staphylococci in the capital area of Finland is comparable with many other countries in Europe. Resistance in indicator bacteria, E. coli and enterococci, was low. PMID- 15535086 TI - Endocrine profiles, haematology and pregnancy outcomes of late pregnant Holstein dairy heifers sired by bulls giving a high or low incidence of stillbirth. AB - The high incidence of stillbirth in Swedish Holstein heifers has increased continuously during the last 15 years to an average of 11% today. The pathological reasons behind the increased incidence of stillbirth are unknown. The present experiment was undertaken to investigate possible causes of stillbirth and to study possible physiological markers for predicting stillbirth. Twenty Swedish Holstein dairy heifers sired by bulls with breeding values for a high risk of stillbirth (n = 12) (experimental group) and a low risk of stillbirth (n = 8) (control group, group B) were selected based on information in the Swedish AI-data base. The experimental group consisted of 2 subgroups of heifers (groups A1 and A2) inseminated with 2 different bulls with 3.5% and 9% higher stillbirth rates than the average, and the control group consisted of heifers pregnant with 5 different bulls with 0%-6% lower stillbirth rates than the average. The bull used for group A1 had also calving difficulties due to large calves as compared to the bull in group A2 showing no calving difficulties. The heifers were supervised from 6-7 months of pregnancy up to birth, and the pregnancies and parturitions were compared between groups regarding hormonal levels, haematology, placental characteristics and calf viability. In group A1, 1 stillborn, 1 weak and 4 normal calves were recorded. In group A2, 2 stillborn and 4 normal calves were registered. All animals in the control group gave birth to a normal living calf without any assistance. The weak calf showed deviating profiles of body temperature, saturated oxygen and heart rates, compared with the normal living calves. No differences of the placentome thickness, measured in vivo by ultrasonography were seen between the groups. The number of leukocytes and differential cell counts in groups A1 and A2 followed the profiles found in the control group. In group A1, a slight decrease of oestrone sulphate (E1SO4) levels was found in the animal delivering a stillborn calf from the first 24-h blood sampling at 6 weeks to the second at 3 weeks prior to delivery, while the levels of E1SO4 at both periods in the animal delivering a weak calf followed the profile in animals delivering a normal living calf. During late pregnancy and at the time of parturition, the levels of E1SO4 and PAGs in animals delivering a stillborn or weak calf (from group A1) followed the normal profiles found in animals delivering a normal living calf. In group A2, low levels of E1SO4 and pregnancy associated glycoproteins (PAGs) over 24 h at both 3 and 6 weeks prior to parturition (< 1.5 nmol/L) were recorded in animals delivering a stillborn calf. During late pregnancy and parturition, the levels of E1SO4 and PAGs were slightly lower during 30-50 days prior to delivery and increased with a lower magnitude at the time of parturition. In conclusion, our results indicate that the aetiology behind stillbirth varies depending on the AI-bulls used and is associated with dystocia or low viability of the calves. Deviating profiles of oestrone sulphate (E1SO4) and pregnancy associated glycoproteins (PAGs) in animals delivering a stillborn calf not caused by dystocia were observed, suggesting placental dysfunction as a possible factor. The finding suggests that the analyses of E1SO4 and PAGs could be used for monitoring foetal well-being in animals with a high risk of stillbirth at term. PMID- 15535087 TI - Hepatic cobalt and copper levels in lambs in Norway. AB - Cobalt and copper concentrations were measured in 599 lamb livers collected at slaughter from 58 sheep flocks in 6 different parts of Norway in 1993. Information about pasture, additional feeding and mineral supplements in the flocks was obtained through a questionnaire. Average hepatic levels of cobalt in the lamb flocks varied from < 0.003 to 0.22 microg/g ww, and of copper from 5 to 240 microg/g ww. Flocks with deficient or marginal cobalt status were found in all parts of southern Norway, but primarily in the west and south-west. Some flocks with marginal copper status were found in the south-west, while flocks with signs of excessive hepatic copper concentrations were found mainly in inner parts of central and northern Norway. Hepatic copper concentrations were significantly higher in lambs that had grazed mountain pastures than in those that had grazed lowland pastures in the summer. PMID- 15535088 TI - Acute phase proteins in response to Dictyocaulus viviparus infection in calves. AB - Three experiments were carried out to examine the acute phase response, as measured by the acute phase proteins (APP) haptoglobin, serum amyloid A (SAA) and fibrinogen, in calves infected with lungworm, Dictyocaulus vivparus. In addition, eosinophil counts were analysed. Three different dose models were used in 3 separate experiments: 1) 250 D. viviparus infective third stage larvae (L3) once daily for 2 consecutive days, II) 100 D. viviparus L3 once daily for 5 consecutive days, and III) 2000 L3 once. All 3 dose regimes induced elevated levels of haptoglobin, SAA and fibrinogen, although there was considerable variation both between and within experiments. A significant increase was observed in all 3 APP at one or several time points in experiment I and III, whereas in experiment II, the only significant elevation was observed for fibrinogen at one occasion. The eosinophil numbers were significantly elevated in all 3 experiments. The results show that lungworm infection can induce an acute phase response, which can be monitored by the selected APP. Elevated APP levels in combination with high numbers of eosinophils in an animal with respiratory disease may be used as an indicator of lung worm infection, and help the clinician to decide on treatment. However, high numbers of eosinophils and low levels of APP do not exclude a diagnosis of lungworm. Thus, lungworm infection may not be detected if measurements of APP are used to assess calf health in herds or individual animals. PMID- 15535089 TI - Influence of cereal non-starch polysaccharides on ileo-caecal and rectal microbial populations in growing pigs. AB - The effect of cereal non-starch polysaccharides (NSP) on the gut microbial populations was studied in 5 growing pigs between 39-116 kg body weight according to a Latin square design. The diets were composed to contain different NSP levels. The control diet had a normal NSP content (139 g/kg dry matter (DM)), 2 diets had a low total amount of NSP (95 and 107 g/kg DM) and 2 diets had a high amount of total NSP (191 and 199 g/kg DM). Furthermore, one of the diets within each category had a content of insoluble NSP similar to the control diet and one had a high content of insoluble NSP. Samples were collected from the ileum, via intestinal post valve T-caecum (PVTC) cannulas surgically inserted at the ileo caecal ostium, and from the rectum. The total microbial flora of the ileal samples were analysed for by defining base pair length with terminal restriction fraction length polymorphism (T-RFLP). The microbial diversity of the coliform flora of the ileal and rectal samples were defined by biochemical fingerprinting. It was observed that many terminal restriction fragments (TRFs) disappeared when new diets were introduced and that some characteristic TRFs were found in the high and low NSP diets, respectively. Both the total gut microflora and the coliform flora were influenced by the dietary NSP content. PMID- 15535090 TI - Commercially available antibodies to human tumour necrosis factor-alpha tested for cross-reactivity with ovine and bovine tumour necrosis factor-alpha using flow cytometric assays. AB - A thorough understanding of the immune system, including the role of different cytokines, during inflammatory diseases in ruminants could lead to the development of new diagnostic methods and treatments. Tumour necrosis factor alpha (TNF-alpha) is an important cytokine in the onset of the inflammatory responses. Unfortunately, the number of studies on cytokines, like TNF-alpha, in ruminants is limited due to a lack of species-specific reagents. As cytokines have remained rather conserved during evolution, cross-reactivity between animal species may occur. Therefore, the aim of the present study was to investigate 5 commercially available antibodies against human TNF-alpha for their ability to cross-react with ovine and/or bovine TNF-alpha, using a bead-based flow cytometric method. Two of the antibody clones (Mab 11 and 6401.1111) showed cross reactivity with ovine recombinant TNF-alpha in concentrations above 2.5 ng/ml. However, none of the antibodies detected TNF-alpha in bovine milk, or serum containing known concentrations of bovine TNF-alpha, as earlier determined with ELISA. The results could be due to inability of the antibodies to cross-react between species, but quenching of the signal by matrix proteins might also have lowered the response. PMID- 15535091 TI - Evaluation of clinical and laboratory variables as prognostic indicators in hospitalised gastrointestinal colic horses. AB - The present prospective study included 106 horses referred to the Department of Large Animal Sciences, The Norwegian School of Veterinary Science, as non responders to the initial colic treatment in general practise. In 14 of these cases a required surgical treatment was not performed due to economical or other reasons and were excluded from the study. Clinical and laboratory data were obtained at the arrival in the hospital. The outcome for all analyses was survival/ non-survival. A multivariable logistic regression was performed. The analyses were used in medically (46 horses) and surgically treated cases (46 horses) separately. The same analyses were also run for all 92 horses in a simulated "field" situation, where only clinical variables and D-dimer values were included. The fraction of survivors was 78% in the medical and 48% in the surgical cases. In total 63% of the horses survived. In the final multivariable logistic regression model packed cell volume (PCV) was the only important predictor for medically treated cases, and heart rate and presence of hyperaemic or cyanotic mucous membranes were the predictors in the surgically treated cases as well as in the simulated "field" situation. In conclusion, traditional variables as heart rate, mucous membranes and PCV were the important predictors for the outcome in hospitalised colic cases. PMID- 15535092 TI - Prediction of vitamin A, vitamin E, selenium and zinc status of periparturient dairy cows using blood sampling during the mid dry period. AB - Vitamins A and E, and the trace elements selenium (Se) and zinc (Zn) are essential for the health and performance of dairy cows. Their concentrations often decrease around calving and extra supplementation is sometimes recommended at that time. However, the need for this varies, for example depending on quantity and quality of feedstuffs in the diet. The aim of this study was to measure the concentrations of serum vitamin A (S-vit A) and vitamin E (S-vit E), plasma Se (P-Se) and serum Zn (S-Zn) in blood samples taken at several time points from one month before to one month after calving, and to evaluate if a blood sample taken during the mid dry period can accurately predict the blood concentration at calving and early lactation. Dairy cows on 3 different feeding regimens during the dry period were included in the study. A significant decrease in the concentrations of S-vit A and S-vit E, and S-Zn, was observed at calving, and P-Se was significantly lower during the dry period and at calving than in early lactation. The blood concentrations of S-vit E and P-Se in the mid dry period significantly predicted the occurrence of values considered marginal or deficient at the time of calving. The data indicate that a mid dry period concentration of > or = 5.4 mg/l of S-vit E and > or = 0.09 mg/l of P-Se will result in a 90% chance that the cow stays above marginal levels at calving given that a feed of the same quality is offered. PMID- 15535093 TI - The parasitological quality of the sandboxes of public playgrounds in Oulu, Finland. PMID- 15535094 TI - The dental facility of the future. PMID- 15535095 TI - Spa dentistry--an accessory not an identity. PMID- 15535096 TI - The integral dental practice; building a "custom" office; managing part-time employees. PMID- 15535097 TI - The magic of effective communication. PMID- 15535098 TI - Business overhead expense insurance. PMID- 15535099 TI - A brief history of the Indiana Dental Practice Act and jury duty exemption. PMID- 15535100 TI - Indiana Medicaid web-based services. PMID- 15535102 TI - 1979-2004: what happened? PMID- 15535101 TI - S. Edith Davis, DDS (1890-1984): pioneer Indiana periodontist. PMID- 15535103 TI - [Aortic distensibility indices evaluated by transoesophageal echocardiography in patients with coronary artery disease or aortic valve stenosis]. AB - INTRODUCTION: The authors examined the aortic distensibility indices evaluated by transoesophageal echocardiography (TEE) in patients with ischaemic heart disease and in aortic stenosis cases with normal epicardial coronary arteries. PATIENTS AND METHODS: 126 consecutive patients with chest pain were enrolled into the study with the following results: 17 patients showed normal epicardial coronary arteries, 23 patients had non-significant coronary artery disease, 44 patients had significant one-vessel disease and 42 patients had significant multivessel disease. Their results were compared to 16 aortic stenosis cases but with negative coronary angiograms. All patients underwent transthoracic and transoesophageal echocardiography and coronary angiography. During a complete TEE the following data were recorded: systolic and diastolic blood pressure and heart rate, systolo-diastolic aortic diameters and intimo-medial thickness. According to these data elastic and Young's moduli were calculated. RESULTS: The indices of aortic distensibility were significantly increased (the aortic distensibility was decreased) in patients with coronary artery disease (CAD) as compared to cases with normal epicardial coronary arteries independently the number of affected vessels. In cases of aortic stenosis but with normal epicardial coronary arteries, the aortic distensibility indices were similarly increased as in patients with CAD. CONCLUSIONS: There is a considerable stiffness of the descending aorta of patients with CAD than in cases with normal epicardial coronary arteries. The aortic distensibility of aortic stenosis patients with negative coronary angiograms are similarly decreased that in patients with CAD. PMID- 15535104 TI - [Oxidative stress and liver disease]. AB - Several papers have been published on the important role of oxidative stress on living cells and cell responses e.g. apoptosis and necrosis, which leads to cell death. At the same time mild oxidative stress can modulate signal transduction cascades and redirect gene expression, and influence many cellular responses, e.g. proliferation, differentiation, reproduction. Regulations of the cell cycle depend on intracellular redox state. Critical steps in the signal transduction cascade are sensitive to oxidative stress and antioxidants. Heavy metal accumulation in higher concentration may inhibit enzyme activities influence the acute phase protein synthesis and gene expression, as well as the pro-oxidant and antioxidant forms of scavenger molecules. Polyphenols and flavonoid type antioxidants may influence the signal transduction routes as well. Ethanol inducible cytochrome P450 2EI isoenzyme oxidise ethanol and acetaldehyde and numerous potentially toxic xenobiotic and produce toxic oxygen free radicals, which are implicated in the pathogenesis of alcoholic liver diseases. Natural preparations, e.g. tea infusions contains trace elements and polyphenol type antioxidants in high concentration, therefore may influence the redox homeostasis, and especially dangerous with interaction of other medicines and alcohol. PMID- 15535105 TI - [The allergic and other side effects of non-steroid antiinflammatory drugs and gold salts]. AB - The wider usage of non steroid antiinflammatory drugs (NSAIDs) raises the significance of their side effects. The discovery of the two different cyclo oxygenases (COX 1 and COX-2) led to the incorporation of more selective enzyme inhibitors into the therapeutic tools against disorders with pain and inflammation, in order to minimize the frequency of the side effects. Selective COX-2 inhibitors are well tolerated by most of the patients with a history of sensitivity against classical NSAIDs. The well-known gastrointestinal side effects (ulcers, bleedings) are much less frequent in the case of selective COX-2 inhibitors in comparison with non-selective COX inhibitors. However, the lack of "healing" prostaglandins as an effect of COX-2 antagonism may prevent the improvement of existing ulcers. In addition almost all other organs have been found to be affected in COX-2 knockout mice (COX-2 paradoxon). Hepatotoxicity is usually rare, its reason is most probably idiosyncrasy. Persistent nephropathy can be worsened by the inhibition of COX-2, however normal renal functions have not been changed in humans using selective COX-2 inhibitors. Authors' registry consists of 1000 patients with a history of suspected drug-allergy, during a 15 years' period. Approximately 30% of the cases have been connected with NSAIDs and with antirheumatic drugs. Because of functional similarities gold salts, proved suitable for the treatment of juvenile rheumatoid arthritis (JRA) and of osteoarthritis (OA) were included as well. Besides rheumatologic applications the second most common indication for these drugs was pain and/or fever. Among cutaneous symptoms intolerance was present at a relatively low frequency--as salicylates had not been taken into consideration. Next to salicylates the most frequent side effects were caused by pyrazolon derivates. Urticaria and angioedema were the most frequently observed symptoms on the skin--our observations are in accordance with other publications. CONCLUSIONS: Non selective NSAIDs show a mixed effect of inhibitions of COX-1 and COX-2. The most important indications of modern selective COX-2 inhibitors are: 1. Prevention of the gastrointestinal side effects 2. Avoidance of cross-sensitivity against non selective NSAIDs. PMID- 15535106 TI - [Beta-adrenergic receptor blockers in clinical practice]. AB - Discovery of beta receptor system and the class of beta-adrenergic receptor antagonists in the last century has been one of the most important developments in cardiovascular pharmacotherapy. A non-cardioselective beta blocker (propranolol) was introduced in the therapy in the early 1960s. Later, the second generation beta1 selective blockers and finally the third generation blockers (with vasodilator property) appeared on the market. Beta-adrenergic receptor antagonists have become widely used in cardiovascular therapy for the last decades. They were introduced for the treatment of angina pectoris and arrhythmias and later hypertension and there has been a continuing progress in the treatment of several heart diseases (including congestive heart failure which was contraindicated earlier). The recent clinical indications and practical considerations of beta-blockers are summarized in this review. PMID- 15535107 TI - [Treatment of neuralgias]. PMID- 15535108 TI - [A. Ullmann, M. Kaposi and J. A. Fournier]. PMID- 15535109 TI - [Hungarian physicians in the USA]. PMID- 15535110 TI - Antitussive effect of diltiazem in experimental conditions. AB - BACKGROUND: Cough is the most important defence reflex of the airways. Little information exists about participation of the ion channels in the mechanism of the cough reflex. OBJECTIVES: The aim of the study was to investigate the effect of diltiazem administration on the sensitivity of the mechanically induced cough reflex in experimental conditions. METHODS: Mechanical stimulation of the laryngopharyngeal and tracheobronchial area of the airways in non-anaesthetized cats was used in this experiment. The number of cough efforts, and the frequency and intensity of the cough attack were utilized as evaluated parameters. RESULTS: Perorally administered diltiazem (30 mg/kg b.w.) caused a statistically significant decrease of measured cough parameters. Cough - suppressing activity of diltiazem was lower than codeine, but higher than dropropizine and prenoxdiazine. CONCLUSION: Experimental results showed antitussive activity of the calcium-channel blocker diltiazem and suggested the role of voltage-gated calcium channels of type L in mechanism of the cough reflex. (Fig. 4, Ref. 15.). PMID- 15535111 TI - Metformin versus sibutramine in the treatment of hyperinsulinemia in chronically anovulating women. AB - Aim of this study was to compare the effects of metformin and a body weight reduction regimen using sibutramine on insulinemia, insulin sensitivity, and ovarian function in women with anovulatory cycles or infertility. 30 women with anovulatory cycles and hyperinsulinemia were treated with metformin and 15 anovulating women with obesity were treated with sibutramine in combination with a caloric restriction diet and physical exercise. In the metformin group there was a mild decrease of the body mass index (BMI), a decrease of fasting and stimulated insulinemia (I0, p < 0.05, I120, p < 0.01), a significant reduction of insulin resistance calculated as index FIRI (p < 0.05), serum LH (p < 0.05) and testosterone levels (p < 0.05). There was an improvement of menstrual cycles in 21 (70 %) of women, and 6 of them became pregnant. In the sibutramine group we found a significant decrease of BMI (p < 0.01), waist circumference (p < 0.01), fasting and stimulated insulinemia (p < 0.05, p < 0.01) and a significant improvement of insulin sensitivity (FIRI, p < 0.01). However, the levels of FSH, LH, and testosterone were not significantly changed. There was a significantly greater reduction of insulin levels and FIRI after sibutramine treatment compared with metformin treatment, while the changes of LH were not signifcantly different. Testosterone was changed more after metformin therapy. We conclude that although the body weight reduction using sibutramine has a more pronounced effect on insulinemia and insulin sensitivity, metformin may be more effective in the prompt restoration of ovarian function. (Tab. 3, Ref. 24.). PMID- 15535112 TI - Calcitriol load test in postmenopausal osteoporosis: a pilot study. AB - BACKGROUND: Calcium and vitamin D balance is of critical importance in both, prevention and treatment of osteoporosis. The evaluation of this balance is difficult under steady state conditions, therefore the effect of a single oral calcitriol load in postmenopausal women with osteopenia/osteoporosis was used. METHODS: Mineral and hormone concentrations were determined under basal conditions and after the administration of a single 0.5 microg calcitriol dose (Rocaltrol, Roche). RESULTS: Single oral calcitriol dose was administered to 36 postmenopausal women. Increased calciuria (p < 0.001) and calcium fractional excretion (p < 0.001) 24 hours after drug administration indicated kidney participation in calcium homeostasis. Urinary calcium excretion after the calcitriol load correlated with basal urinary calcium excretion (r = 0.772; p < 0.001). On the basis of calciuric response, women could be separated to responders and non-responders; in responders, increase in calciuria was >1.0 mmol/d, while in non-responders < 1.0 mmol/d (p < 0.001). Neither plasma calcium nor plasma 25-hydroxycholecalciferol concentrations increased after the calcitriol load. Both basal (r = -0.361; p < 0.03) and calcitriol stimulated urinary calcium excretion correlated inversely with age (r = -0.425; p < 0.01). No significant changes in measured mineral and hormone parameters were found in women with osteopenia in comparison to women with osteoporosis, with the exception of plasma intact parathormone concentration, which was significantly higher in osteoporotic women (p < 0.05) and significantly decreased (p < 0.001) after the calcitriol load. CONCLUSION: Postmenopausal women suffered from calcium and vitamin D imbalance which could be elucidated by a simple calcitriol test. The reference range, significance and differentiation from other abnormalities are to be defined in an extensive study. (Tab. 4, Ref. 19.). PMID- 15535113 TI - Is there any role for prevention strategies for colorectal cancer other than population-based screening? AB - Colorectal carcinoma is a growing medical problem. Prevention represents the most effective approach for reducing the incidence and mortality. Population-based screening with subsequent colonoscopic polypectomy is the most common strategy. There are, however, other approaches increasing the portfolio of available measures. These include the special care of groups with an increased risk for CRC based on family or medical history and various chemopreventive strategies. Family history is a simple method to identify a person with increased risk of CRC. There are two groups of familial CRC: monogenetic hereditary syndromes such as FAP and HNPCC and hereditary predispositions with sporadic CRC. Recent advances in genetic tests and tailored surveillance strategies are able to decrease the morbidity and mortality in these groups. Therefore a wider recognition of family history as risk indicator of CRC should be encouraged. Chemoprevention is a very promising concept for both primary and secondary prevention of CRC. Although definite evidence is difficult to provide a number of studies suggest a role for nutritional interventions and/or chemoprevention with plant phytosterols, fiber, selenium, calcium, probiotics or COX2 inhibitors as putative chemopreventive strategies. (Tab. 4, Ref. 70.). PMID- 15535114 TI - New progression in the study of protective properties of resveratrol in anticardiovascular disease. AB - Phytoantitoxin resveratrol, which belongs to polyphenol, is a kind of phytoestrogen, and protects the cardiovascular system against ischemic reperfusion injury and promotion of vasorelaxation, protects and maintains the intact endothelium, exhibits antiatherosclerotic properties inhibits the LDL oxidation, suppress the platelet aggregation and exhibits estrogen like action. The purpose of this article is to review the mechanisms of about mentioned effects. (Ref. 30.). PMID- 15535115 TI - Metallothioneins and zinc: their functions and interactions. AB - The impact of environmental and occupational contamination on living organisms has been an interesting topic for scientific and lay public for a long time. The synthesis of metallothioneins is one of the protection strategies, by which organisms protect themselves from metal-induced toxicity. There are four known isoforms of metallothioneins. This article summarizes the role of the widely expressed mammalian metallothionein-1 and metallothionein-2 isoforms in zinc homeostasis, apoptosis, and as novel potential marker of ageing. The metallothionein-3 and metallothionein-4 have been discovered to possess distinct functional properties. The properties of metallothioneins and their interplay are likely to be essential for the still elusive biological function of these proteins. (Ref. 49.). PMID- 15535116 TI - Stereotactic evacuation of spontaneous infratentorial hemorrhage with monitoring of intracerebral pressure. AB - INTRODUCTION: Based on our experience with stereotactic evacuation of spontaneous supratentorial hematomas this method has also been used for evacuation of spontaneous infratentorial hematoma by the transtentorial approach. MATERIAL AND METHOD: The authors present a series of 6 patients with spontaneous intracerebral hematomas evacuated by the frame-based stereotactic technique, with monitoring of intracranial pressure (ICP) during the stereotactic evacuation. This method was indicated in patients with stable neurological status according to Glasgow Coma Scale (GCS), more than 10. The frame-based stereotaxy with the Riechert-Mundinger apparatus with CT localisation of target and optimal trajectory was used. RESULTS: The presented values after stereotactic evacuation show reduction of the initial intraparenchymal ICP in all patients to values less than 20 mmHg. CONCLUSION: The measuring of the ICP and the analysis of dynamic changes during stereotactic evacuation suggest that this procedure can significantly reduce the ICP performed in connection with ICH and we believe that our results can improve management of patients with spontaneous infratentorial and supratentorial intracerebral hematoma. (Fig. 1, Ref. 21.). PMID- 15535117 TI - Venous thrombosis in the surgical department. AB - Currently, there are no standardized guidelines aimed to the prevention of DVT. Thus we recommend the introduction of a standardized scoring system that would ensure the effective prevention of DVT. (Tab. 6, Ref. 5.). PMID- 15535118 TI - Does exogenous glycinebetaine affect antioxidative system of rice seedlings under NaCl treatment? AB - The effect of exogenously applied glycinebetaine (GB) on the alleviation of damaging effects of NaCl treatment was studied in view of relative water content (RWC), malondialdehyde content, and the activity of some antioxidant enzymes in two rice (Oryza sativa L.) cultivars differing in salt tolerance (salt-tolerant Pokkali and--sensitive IR-28), comparatively. Both cultivars took up exogenously applied GB through their roots and accumulated it to considerable levels. Leaf RWC of both cultivars under salt treatment showed an increase with GB application. The activities of superoxide dismutase (SOD), ascorbate peroxidase (AP), catalase (CAT), and glutathione reductase (GR) increased in leaves of Pokkali, but peroxidase (POX) activity decreased under salinity. In IR-28, the activities of SOD, AP and POX increased, whereas CAT and GR decreased upon exposure to salt treatment. When compared to the salt-treated group alone, GB application decreased the activities of SOD, AP, CAT, and GR in Pokkali, whereas it increased the activities of CAT and AP in IR-28 under salinity. However, the activity of POX in IR-28 under salinity showed a decrease with GB application compared to the NaCl group. In addition, lipid peroxidation levels of both cvs. under salt treatment showed a decrease with GB treatment. Therefore, we conclude that GB protects both rice seedlings from salinity-induced oxidative stress. PMID- 15535119 TI - Light dependency of resistance to ionizing radiation in Euglena gracilis. AB - The resistance of Euglena gracilis strains Z (wild type) and SM-ZK (chloroplast deficient mutant) to ionizing radiation was investigated. The colony forming ability of E. gracilis strain Z was higher than that of strain SM-ZK after 60Cogamma-irradiation. For both strains, the resistance of light-grown cells was higher than that of dark-grown cells, suggesting that the light conditions during culture contribute to the radiation resistance of E. gracilis. The comet assay showed that the ability of rejoining DNA double-strand breaks (dsb) was much higher in the light-grown cells. These results suggest that E. gracilis possesses a light-induced repair system to cope with DNA dsb. PMID- 15535120 TI - Developing embryos of Sesbania sesban have unique potential to photosynthesize under high osmotic environment. AB - This study has been carried out to investigate the photosynthetic activities in developing embryos of Sesbania sesban under a highly osmotic environment. In S. sesban, the embryo turns green/chlorophyllous at the early heart shape stage. Interestingly, despite being deeply embedded within the supporting tissues (several layers of pod wall, seed coat and endosperm) and developing in a highly osmotic environment, the growing embryo of the developing seed showed the presence of various components of photosynthetic machinery besides being chlorophyllous. The shade-adaptive nature of the photosynthetic machinery of the embryo is evident from (a) low chlorophyll a/b ratio, (b) photosystem (PS) II attaining maximal activity at low photon flux density and (c) lesser plastoquinone pool. The photosynthetic potential of the growing embryo seems to contribute towards seed filling as it has the potential not only to harvest light energy but also to fix CO2 as efficiently as other photosynthetic parts of S. sesban. In fact, ribulose-1,5 bisphosphate carboxylase purified from embryos manifested subunit composition similar to that of leaves. The PS II activity in leaves, cotyledonary leaves and pod wall declined sharply with increase in the level of NaCl and sucrose above 150 and 300 mmol, respectively. Amazingly, PS II activity in developing embryos was maximal in the presence of 250 mmol NaCl or 500 mmol sucrose and remained high even when NaCl and sucrose levels were increased to 500 and 1000 mmol, respectively. We hypothesize that the developing embryos have some factor(s) which protect(s) the photosynthetic machinery in an environment of high osmotic strength. PMID- 15535121 TI - Causes and consequences of high osmotic potentials in epiphytic higher plants. AB - Past reports of the water relations of epiphytes, particularly bromeliads, indicate that tissue osmotic potentials in these tropical and subtropical plants are very high (close to zero) and are similar to values for aquatic plants. This is puzzling because several ecophysiological studies have revealed a high degree of drought stress tolerance in some of these epiphytes. The goal of this study was two-fold: (1) to increase the number of epiphytic taxa sampled for tissue osmotic potentials; and (2) to explain the apparent discrepancy in the significance of the tissue water relations and tolerance of drought stress in epiphytes. Tissue osmotic potentials of 30 species of epiphytic ferns, lycophytes, and orchids were measured in a subtropical rain forest in northeastern Taiwan. Nearly all values were less negative than -1.0 MPa, in line with all previous data for epiphytes. It is argued that such high osmotic potentials, indicative of low solute concentrations, are the result of environmental constraints of the epiphytic habitat on productivity of these plants, and that low rates of photosynthesis and transpiration delay the onset of turgor loss in the tissues of epiphytes such that they appear to be very drought stress tolerant. Maintenance of photosynthetic activity long into drought periods is ascribed to low rates of transpiration and, hence, delayed tissue desiccation, and hydration of the photosynthetic tissue at the expense of water from the water storage parenchyma. PMID- 15535122 TI - Modulation of pigment profiles of Calothrix elenkenii in response to environmental changes. AB - Cyanobacteria are versatile tetrapyrrole synthesizers that can regulate their tetrapyrrole content and composition in response to environmental signals. The present investigation analyses the interplay between light and dark regimes (continuous light, light-dark cycles (16:8) and continuous darkness) and aerobic, air-tight, and anaerobic environments (argon-enriched), on the relative composition of various pigments and growth attributes of Calothrix elenkenii as a prelude to exploiting C. elenkenii's bioindustrial potential as a source of pigments. Incubation in an anaerobic environment stimulated hormogonia formation and induced colouration/thickening of cells. Aerobically grown cultures of Calothrix, under continuous illumination produced the maximum amount of total phycobiliproteins and sugars, although chlorophyll accumulation and nitrogenase activity were highest in the light-dark environment. However, the beta-carotene content was observed to vary under anaerobic conditions with different light-dark regimes. This C. elenkenii strain can be a valuable source of pigments under optimized environmental conditions. PMID- 15535123 TI - Effects of irrigation and air humidity preconditioning on water relations, growth and survival of Rosmarinus officinalis plants during and after transplanting. AB - The effect of different irrigation and air humidity conditioning treatments on the morphological and physiological responses of Rosmarinus officinalis in nursery conditions was investigated in order to evaluate the degree of hardening resulting from these conditions. Rosmarinus officinalis seedlings were pot-grown during 4 months in two greenhouses (nursery period), in which two irrigation treatments were used (control and deficit). In one of these greenhouses, air humidity was controlled using a dehumidifying system (low humidity), in the other greenhouse the air conditions were not artificially modified (control humidity). After the nursery period, the plants of all treatments were transplanted and well watered (100% water holding capacity for 1 month, transplanting period). After this period, they received no water (establishment period). At the end of the nursery period it was seen that deficit irrigation had altered the morphology of the R. officinalis plants by reducing plant height, stem diameter, leaf area, total dry weight, and root length, while humidity influenced the parameters related with plant water relations. Low air humidity and deficit irrigation induced tissue dehydration and lower stomatal conductance values (gs). The plants subjected to deficit irrigation developed leaf osmotic adjustment, which was maintained during the transplanting period. At that time, the plants that had been exposed to deficit irrigation and low humidity showed efficient stomatal regulation (lower gs values). After transplanting and during the establishment period, these plants showed a better water status (higher psil and gs values). Their post-planting survival rate improved as a result of acclimation processes. PMID- 15535124 TI - Reactive oxygen species and nitric oxide affect cell wall metabolism in tobacco BY-2 cells. AB - The effects of hydrogen peroxide (H2O2), nitric oxide (NO), and a combination of both on the metabolism of cell wall polysaccharides were studied in tobacco (Nicotiana tabacum L.) cv Bright Yellow 2 (BY-2) suspension cultured cells in the presence of D-[U-14C]glucose or D-[U-14C]galactose as radioactive tracers. We found that the radiolabelling of newly synthesised total cell wall polysaccharides (pectins, hemicelluloses and alpha-cellulose), buffer-soluble polysaccharides, and membrane-associated polysaccharides decreased under the influence of exogenous systems generating H2O2 and NO. However, when the total amount of newly synthesised cell wall polysaccharides was calculated as a percentage of the total cellular radioactivity (ethanol-soluble pool plus the homogenate of ethanol-insoluble material), all treatments showed negligible effects in the presence of D-[U-14C]glucose or D-[U-14C]galactose as tracers. This occurred because the treatments generating H2O2, NO and H2O2 plus NO caused a marked decrease in the concentration of the ethanol-soluble pool as well as in the total radioactivity found in the homogenate of the ethanol-insoluble material. Most of the radioactivity taken up by the cells was evolved as 14CO2 during the respiratory processes. A qualitative and quantitative characterisation of the ethanol-soluble pool showed that radioactive UDP-sugars in BY-2 suspension cultured cells were differentially reduced by all treatments. Therefore, the decrease of the newly synthesised cell wall polysaccharides seems to be strictly dependent on the reduction of the UDP-sugars pool. PMID- 15535125 TI - Senescence-dependent degradation of Lhcb3 is mediated by a thylakoid membrane bound protease. AB - A novel proteolytic activity integrally associated with barley thylakoid membranes has been discovered and characterized. This enzymatic activity mediates senescence-dependent degradation of Lhcb3, one of the apoproteins of the major light-harvesting chlorophyll a/b protein complex of photosystem II. Once senescence of barley leaves is initiated by detachment and dark incubation, the degradation of Lhcb3 can proceed and be followed in vitro in an experimental system composed of thylakoids isolated from senescing leaves incubated in darkness in suitable medium at 25 degrees C. The protease involved is present in its active form and Lhcb3 is susceptible for proteolytic attack already in fresh leaves, although Lhcb3 degradation does not take place unless undefined extrinsic membrane proteins protecting Lhcb3 are removed in a senescence-dependent manner. It is thus concluded that senescence-dependent Lhcb3 degradation is regulated at the substrate availability level. The protease involved is ATP stimulated, has an optimum activity at pH 7.8, and requires 3 mM added Mg2+ (replicable by micromolar doses of Zn2+) for its proper activity. Studies using typical inhibitors of various classes of proteases indicate that the enzyme is a metalloprotease with disulfide linkage essential for its activity. Micromolar doses of Zn2+ were demonstrated to restore the activity of Lhcb3-degrading enzymes abolished by an ethylenediaminetetraacetic acid pretreatment of the thylakoids and it is inferred that the protease involved is a zinc-binding metalloprotease. Mg2+ was shown to be able to partially replace zinc as the bound ion. PMID- 15535126 TI - Expression of a sweet cherry DREB1/CBF ortholog in Arabidopsis confers salt and freezing tolerance. AB - Dehydration responsive element binding protein 1 (DREB1)/C-repeat binding factor (CBF) induces the expression of many stress-inducible genes in Arabidopsis. We have previously reported the identification of three DREB1/ICBF homologs from sweet cherry (Prunus avium). To identify the function of these homologs, one of the genes, CIG-B, was transformed into Arabidopsis. In one of the transgenic plant lines, the DREB1/CBF target gene cor15a was induced in the absence of stress treatment. The cor15a-overexpressing transgenic plant exhibited mild growth retardation and had greater salt and freezing tolerance than did the wild type and the transgenic lines in which cor15a was not induced. These results suggest that this sweet cherry DREB1/CBF homolog has a function similar to that of DREB1/CBF. PMID- 15535127 TI - Engineered sorbitol accumulation induces dwarfism in Japanese persimmon. AB - A cDNA encoding sorbitol-6-phosphate dehydrogenase (S6PDH), which is a key enzyme in sorbitol biosynthesis in Rosaceae, was introduced into the Japanese persimmon (Diospyros kaki) to increase the environmental stress tolerance. Resultant transformants exhibited salt-tolerance with dwarfing phenotypes. Therefore, we studied two transgenic lines to understand the physiological mechanism of this dwarfism: lines PS1 and PS6 accumulated high and moderate levels of sorbitol, respectively. The average length of shoots was significantly shorter as compared with the wild-type in line PS1, while no such decrease was observed in line PS6. The myo-inositol and glucose 6-phosphate (G6P) contents were measured in the transgenic lines because previous work with tobacco transformed with S6PDH had suggested that growth inhibition was due to depletion of these metabolites. Although the myo-inositol content was decreased in PS1 plants, the decrease was much smaller than that observed in transgenic tobacco that accumulates sorbitol. The G6P contents were the same in PS1 plants and phenotypically normal PS6 plants. The level of indole-3-acetic acid (IAA), which affects stem elongation, in line PS1 was similar to the levels in the other lines. A decrease in gibberellin (GA) content generally induces dwarfism in plants. However, GA was not decreased in PS1 plants compared with wild-type or control plants. Therefore, we focused on sorbitol accumulation as the most remarkable feature of PS1 plants. As one possibility, the observed growth inhibition was likely caused by an osmotic imbalance between the cytosol and vacuole. PMID- 15535128 TI - DNA chip technology in brain banks: confronting a degrading world. AB - DNA microarray technology is based on the principle of hybridization between 2 complementary strands of nucleic acids, one being fixed into a solid membrane, the other being the sample to analyze. This has resulted in a very powerful method to examine differential gene expression between samples, and has been widely used in the study of tumors. The application of DNA microarray technology to the study of the nervous system has to consider several properties of the nervous tissue: composition of various neuronal types, as well as astrocytes, oligodendrocytes, and microglia; regional and area differences; developmental and age-dependent variations; and functional and pathological status. Moreover, human samples are usually obtained postmortem following variable agonal periods and postmortem delays between death and tissue preservation, which are accompanied by variable RNA degradation. Yet human postmortem nervous tissue stored in brain banks offers a unique opportunity to facilitate material for the study of diseases of the nervous system and to gain direct understanding on the mechanisms of disease. This review analyzes the application of DNA microarray technology to current practice using brain-banked tissues in order to recognize and minimize sub-optimal processing of brain samples and to correct pitfalls due to inadequate procedures. Also discussed are RNA preservation and RNA degradation effects on expression pattern assessments, analysis of individual versus pooled samples, array normalization, types of DNA chip platforms, whole genomic analysis versus specialized chips, and microgenomics. Minimizing RNA degradation and improving detection of resistant RNA in postmortem brain has been considered in detail in order to improve the efficiency and reliability of DNA microarray technology employed in the study of human postmortem nervous tissue. PMID- 15535129 TI - Loss of tumor suppressor in lung cancer-1 (TSLC1) expression in meningioma correlates with increased malignancy grade and reduced patient survival. AB - Meningiomas represent the second most common central nervous system tumor affecting adults. Two of the most frequent early events in meningioma tumorigenesis involve loss of expression of the neurofibromatosis 2 (NF2) and 4.1B genes. Recently, 4.1B was shown to interact with the tumor suppressor in lung cancer-1 (TSLC1) protein, prompting us to examine the expression of TSLC1 in meningiomas. We developed specific anti-TSLC1 antibodies to examine TSLC1 expression in normal human leptomeninges, human meningioma cell lines, and human meningiomas of different pathological grades by Western blot (n = 10) and immunohistochemistry (n = 123). Whereas TSLC1 was expressed in normal human leptomeninges by immunohistochemistry, TSLC1 expression was absent in 3 human malignant meningioma cell lines and markedly reduced or absent in 30% of benign meningiomas by Western blot. Restoration of TSLC1 expression in a TSLC1-deficient human meningioma cell line resulted in reduced cell proliferation. In a series of 123 meningiomas (98 adult and 25 pediatric), TSLC1 expression was absent in 48% of benign (WHO grade I), 69% of atypical (grade II), and 85% of anaplastic (grade III) meningiomas. Moreover, TSLC1 loss was associated with decreased patient survival, within the overall group, and in the atypical meningiomas. Collectively, these results suggest that TSLC1 plays an important role in meningioma pathogenesis. PMID- 15535130 TI - Neuropathologic criteria for diagnosing Alzheimer disease in persons with pure dementia of Alzheimer type. AB - Universally accepted neuropathologic criteria for differentiating Alzheimer disease (AD) from healthy brain aging do not exist. We tested the hypothesis that Bielschowsky silver stained total, cored, and neuritic senile plaques (TSPs, CSPs, and NSPs, respectively), rather than neurofibrillary tangles (NFTs), best discriminate between the 2 conditions using rigorously defined nondemented (n = 7) and AD (n = 35) subjects with no known co-morbidities. We compared lesions in 3 neocortical regions, in hippocampal CA1, and in entorhinal cortex in 19 men and 13 women between 74 and 86 years at death. The Clinical Dementia Rating (CDR) was used to assess degree of cognitive impairment within a year of demise. Neocortical TSP measures provided the highest correlation with expiration CDR: area under the curve (AUC) = 0.986 with 97.8% sensitivity at 90% specificity with an estimated cut-point of 6.0 TSP/ mm2. All SP measures yielded higher estimated AUC and sensitivity for 90% specificity compared to NFTs. Derived TSP cut-points applied to 149 persons with clinical AD regardless of their neuropathologic diagnosis yielded a sensitivity of 97% and specificity of 84% for TSPs in the 3 neocortical areas. Thus cut-points based on both diffuse and neuritic SP in neocortical regions distinguished nondemented and AD subjects with high sensitivity and specificity. PMID- 15535131 TI - Altered P-glycoprotein expression in AIDS patients with HIV encephalitis. AB - Penetrance of anti-retroviral drugs into the CNS depends partly on the activity of P-glycoprotein (P-gp), an ATP-dependent efflux pump involved in restricting entry of lipophilic drugs into the brain. The present study characterizes the patterns of P-gp expression in the brains of AIDS patients and examines its relationship with clinical and neuropathological indicators of HIV encephalitis (HIVE). For this purpose, brain tissue collected at autopsy from 26 subjects with a history of HIV (9 without HIVE; 17 with HIVE) was analyzed. Immunocytochemical staining and Western blot analyses for regional P-gp expression were performed and levels were correlated with neuropathological indicators and with HIV RNA. Double labeling experiments were performed with antibodies against astroglial (GFAP), endothelial (CD31), microglial (CD45) and neuronal (MAP2) cell markers. In the HIVE-negative cases, P-gp immunoreactivity was associated primarily with endothelial cells. HIVE-positive cases showed extensive immunolabeling of astroglial and microglial cells, but relatively less endothelial cell immunolabeling. No neuronal P-gp immunostaining was detected in brain tissue from any cases in the study. In the HIVE-positive cases with extensive astroglial labeling, the most intense immunoreactivity was detected in white matter. A subset of HIVE-positive cases displayed intense P-gp immunostaining of astrocytes closely associated with blood vessels in the cortex. Both the immunocytochemical and Western blot analyses showed a significant correlation between P-gp expression and HIV RNA levels. In conclusion, P-gp immunoreactivity was detected largely in glial cells in tissue from HIVE-positive patients. Furthermore, in HIVE-positive patients, brain viral burden and P-gp levels were significantly higher than those in HIVE-negative patients. Taken together, our data suggest that P-gp may be part of a central pathway mediating viral compartmentalization in the brains of HIV-infected individuals and may play a significant part in HIV disease progression in the brain. PMID- 15535132 TI - Brainstem and basal ganglia lesions in xeroderma pigmentosum group A. AB - Xeroderma pigmentosum group A (XPA) is a hereditary disorder characterized by cutaneous symptoms and progressive neurodegeneration. Since XPA patients exhibit peripheral neuropathy, neuronal deafness, rigidity, dysphagia, and laryngeal dystonia, it is indispensable for investigation of the neurodegeneration to analyze brainstem and basal ganglia lesions clinically and pathologically; we have previously shown the role of oxidative stress in the development of basal ganglia lesions. Here we immunohistochemically examined the expression of neurotransmitters, calcium-binding proteins, and neuropeptides in the brainstem, basal ganglia, and thalamus in 5 XPA autopsy cases. In the brainstem, immunoreactivity for tyrosine hydroxylase, tryptophan hydroxylase, and calbindin D28K was severely reduced throughout the brainstem in all the XPA cases. Nevertheless, the expressions of parvalbumin, substance P, and methionine enkephalin in the brainstem were comparatively preserved; the exception being reduced immunoreactivity for them in the cochlear and dorsal column nuclei in 3 cases. The large cell neurons in the putamen were preferentially reduced, the immunoreactivity for tyrosine hydroxylase reflecting the dopaminergic afferent and efferent pathways was severely affected, and the expression of 3 calcium binding proteins (i.e. parvalbumin, calbindin-D28K, and calretinin) was disturbed in various ways. The expression of substance P and methionine-enkephalin, which are involved in the efferent pathways in the basal ganglia, in the globus pallidus and substantia nigra was spared. It is speculated that the selective damage to the dopamine system in the basal ganglia and the disturbed monoaminergic expression in the brainstem could be related to clinical abnormalities such as the rigidity, laryngeal dystonia, and several neurophysiological changes. Functional analysis of autopsy brains will facilitate clarification of the pathogenesis of the neurodegeneration in XPA. PMID- 15535133 TI - Expression of excitatory amino acid transporter-1 (EAAT-1) in brain macrophages and microglia of patients with prion diseases. AB - The mechanisms of neuronal apoptosis in prion diseases are unclear. Experimental studies suggest that it may result from 2 associated mechanisms: glutamate mediated excitotoxicity and oxidative stress. Recent studies showed that activated macrophages/microglia (AMM) express excitatory amino acid transporters (EAATs) in HIV infection, suggesting that they may play a neuroprotective role by clearing extra-cellular glutamate and producing anti-oxidant glutathione. In order to test this hypothesis in prion diseases, samples from cerebral cortex, striatum, thalamus, and cerebellum from 14 patients with Creutzfeldt-Jakob disease (8 sporadic, 2 familial, 2 iatrogenic, and 2 variant), and 4 with fatal familial insomnia (3 homozygous Met/Met at codon 129 of the PRNP gene, 1 heterozygous Met/Val), and 3 controls were immunostained for EAAT-1, GFAP, HLA DR, CD68, IL-1, caspase 3, and PrP. In prion diseases, EAAT-1 immunopositivity was found in affected areas. Only AMM, interstitial, perivascular, perineuronal (sometimes around apoptotic neurons), or close to reactive astrocytes, expressed EAAT-1. Astrocyte EAAT-1 expression was scarcely detectable in controls and was not detected in prion disease cases. The proportion of AMM expressing EAAT-1 did not correlate with the severity of neuronal apoptosis, spongiosis, astrocytosis, microgliosis, or PrP deposition, but only with disease duration. Occasional EAAT 1 expressing AMM were found in patients with short survival, whereas diffuse EAAT 1 expression by AMM was observed in cases with long survival (24 to 33 months) that most often were heterozygous for Met/Val at codon 129 of the PRNP gene. Our findings suggest that AMM may develop a partial neuroprotective function in long lasting prion diseases, although it does not seem to efficiently prevent neurological and neuropathological deterioration. Whether this neuroprotective function of microglia is the cause or the effect of longer survival needs to be clarified. PMID- 15535134 TI - Loss of heterozygosity at 6q is frequent and concurrent with 3p loss in sporadic and familial capillary hemangioblastomas. AB - Capillary hemangioblastoma is a benign tumor, occurring sporadically or as a manifestation of von Hippel-Lindau (VHL) disease. Inactivation of the VHL gene at 3p25-26 has been demonstrated in all VHL-associated hemangioblastomas. However, the VHL gene has been found to be inactivated in only 20% to 50% of sporadic tumors. So far, no other gene has been reported to be involved in the development of hemangioblastomas. DNA losses at 6q are frequent alterations in hemangioblastomas, as shown by comparative genomic hybridization. We therefore analyzed 15 hemangioblastomas for loss of heterozygosity (LOH) on chromosome 3p and 6q to reveal the frequency of allelic losses and to determine minimal deleted areas. We detected LOH at 6q for one or more markers in 11 (73%) out of 15 cases (in 9 of 11 sporadic and in 2 of 4 VHL-associated tumors). The analyses revealed a minimal 3-megabase (Mb) deleted region at 6q23-24, where 9 of 11 (82%) informative cases showed LOH. LOH at 3p was seen in 14 out of 15 tumors. LOH occurred concurrently at 6q and 3p in 67% of cases. Our data strongly suggests that a tumor suppressor gene located at 6q23-24 is involved in tumorigenesis of hemangioblastomas, in addition to the VHL gene. PMID- 15535135 TI - Downregulation of protein phosphatase 2A carboxyl methylation and methyltransferase may contribute to Alzheimer disease pathogenesis. AB - ABalphaC, a major protein phosphatase 2A (PP2A) heterotrimeric enzyme, binds to and regulates the microtubule cytoskeleton and tau. We have shown that ABalphaC protein expression levels are selectively reduced in Alzheimer disease (AD). Notably, the carboxyl methylation of PP2A catalytic subunit (PP2A(C)) is critically required for ABalphaC holoenzyme assembly, and catalyzed by a specific methyltransferase (PPMT). Here, we provide the first analysis of human PPMT and methylated PP2A(C) in brain regions from AD, non-AD demented, and aged control autopsy cases. Immunoblotting analyses revealed that PPMT protein expression and PP2A(C) methylation levels were quantitatively decreased in AD-affected brain regions. Immunohistochemical studies showed that PPMT was abundant in neurons throughout the cortex in normal control and non-AD demented cases. However, in AD, there was a regional loss of PPMT immunoreactivity that closely paralleled the severity of tau pathology, but not amyloid plaque burden. We propose that the deregulation of PPMT and PP2A methylation/demethylation cycles contributes to AD pathogenesis, by inducing changes in PP2A heteromultimeric composition and substrate specificity. In turn, PP2A dysfunction compromises the mechanisms that control tau, neuronal plasticity, and survival. PMID- 15535136 TI - CGRP peptide and regenerating sensory axons. AB - CGRP peptide, a widely expressed constituent of sensory neurons, plays important roles in nerve function and repair when axons are severed. CGRP synthesis declines, yet peptide nonetheless accumulates in severed axon endbulbs. In this work we explore an apparent selective and ongoing expression of CGRP peptide in regenerative sensory axon sprouts. Following sural nerve crush in rats out to 14 days, regenerating and branching sensory axons had intense and selective expression of CGRP, not associated with endbulbs. Parent L4 and L5 perikarya and axons in the sural nerve proximal to crush, however, did not exhibit such heightened CGRP presence. Instead, back labeling of regenerating axons with fluorogold or diamidino yellow labeled perikarya with reduced CGRP expression. Similarly, ATF-3, a robust marker of axotomized neurons, was associated with reduced, rather than elevated expression of alphaCGRP mRNA. Unexpectedly, however, we identified an enlarged secondary population of intact uninjured neurons, frequently smaller and projecting to the dorsal horn with new and heightened intense CGRP expression but not ATF-3- or tracer-labeled. Distal regenerating sensory axons selectively express CGRP peptide despite reduced perikaryal content, a phenomenon not explained by simple accumulation. Having an injured neighbor neuron, however, may also paradoxically alter how CGRP is expressed in intact neurons. PMID- 15535137 TI - In vivo and in vitro dysferlin expression in human muscle satellite cells. AB - Dysferlin is a protein of the sarcolemma that is mutated in patients with limb girdle muscular dystrophy 2B, Miyoshi myopathy, and distal anterior myopathy. It has been implicated in muscle signaling and sarcolemma repair. To further understand its functional role we studied dysferlin expression in satellite cells (SCs) in normal and pathological human muscle biopsies, as well as in primary cultures of human skeletal muscle. Using immunohistochemistry we detected dysferlin-positive (Dysf+) SCs. Double staining with c-met+, a total SC marker, showed that the number of Dysf+ SCs ranged from 33.7% +/- 4.4% in normal muscle to 68.0% +/- 6.2% in pathological muscles, whereas double staining with MyoD/Dysf showed that all activated SC (MyoD+) were also Dysf+. These results indicate that dysferlin is upregulated in activated SCs. In vitro, immunohistochemistry, semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), and real-time PCR showed that both dysferlin mRNA and protein expression were higher in multinucleated myotubes than in the myoblast stage (p < 0.05). Furthermore, experiments of inhibition of myoblast fusion with amiloride, a type T calcium channel antagonist, showed that dysferlin levels were lower in treated than in non-treated cultures (p < 0.001), demonstrating that dysferlin expression reached peak levels upon differentiation into myotubes. These results and the in vivo findings of dysferlin expression when SCs are activated confirm the involvement of dysferlin in human muscle regeneration/repair and its possible role in fusion events during muscle development. PMID- 15535138 TI - Functional outcomes of reduced hyaluronan in acute vocal fold scar. AB - To examine the functional effects of hyaluronan and collagen alterations in acute vocal fold scar, we injured 15 pig larynges by vocal fold mucosa stripping. At 3, 10, and 15 days after operation, we performed excised larynx experiments to measure phonation threshold pressure (PTP) and vocal economy (an acoustic output cost ratio; OCR), and then performed hyaluronan and collagen assays. Five uninjured larynges were used as excised controls. Hyaluronan was reduced in the scarred vocal folds through 15 days of wound healing. Collagen was increased at day 15. The PTP was increased and OCR was decreased in scarred larynges, indicating decreased vocal efficiency and ease of phonation. Thus, PTP and OCR were sensitive to the biomolecular changes in acute vocal fold scar. Hyaluronan was more susceptible than collagen to acute tissue ultrastructural alterations. These findings may provide a rationale for increasing hyaluronan in acute vocal fold scar to improve postoperative vocal outcomes. PMID- 15535139 TI - Growth factor therapy for vocal fold scarring in a canine model. AB - Vocal fold scarring remains a therapeutic challenge. Hepatocyte growth factor (HGF) has strong antifibrotic activity and has proved to have therapeutic potential in restoration of scar tissues such as liver cirrhosis and lung fibrosis. The present study aimed to clarify the effects of HGF injection into scarred vocal folds in a canine model. Canine vocal folds were stripped unilaterally and treated with intracordal injection of saline solution (sham group), HGF (HGF group), or HGF with cultured autologous normal vocal fold fibroblasts (Fb/HGF group) 1 month after injury. The larynges were harvested 6 months after the initial injury and then subjected to vibratory and histologic examination. The results of vibratory examinations in the excised larynx setup revealed that phonation threshold pressure significantly increased and vocal efficiency was significantly reduced in all treated groups as compared to normal data obtained from normal canine larynges. However, the HGF group presented much better results than both the sham and Fb/HGF groups in terms of mucosal wave amplitude and incidence of vocal fold bowing, glottal incompetence, and phase asymmetry. The histologic data indicated a significant increase of collagen in both the sham and Fb/HGF groups, while normal levels of collagen were found in the HGF group. Tissue contraction of the lamina propria was also observed in both the sham and Fb/HGF groups, but was barely detectable in the HGF group. Although the HGF-treated vocal folds appeared to require more driving forces for vibration, HGF might prevent excessive collagen deposition and tissue contraction and thus reduce the effects of scarring on the vibratory properties of the vocal folds. From these data it is concluded that HGF has considerable potential in the treatment of vocal fold scarring. PMID- 15535140 TI - Tracheal and laryngeal rupture in neonates: complication of delivery or of intubation? AB - Neonatal tracheal or laryngeal rupture is a rare but life-threatening condition that is attributable to traumatic endotracheal intubation or traumatic delivery. We present a review of the literature and 6 new cases of laryngeal or tracheal rupture following complicated delivery. High-risk groups were identified as 1) low-birth weight neonates, for intubation trauma, and 2) extremely high-birth weight neonates with shoulder dystocia, for trauma due to delivery. Two specific types of ruptures can be distinguished. The less-rare type involves a partial anterior rupture in the subglottic area, which can occur after traumatic intubation or traumatic delivery. The rarer type involves a distal circumferential tracheal rupture that gives rise to ventilatory problems, usually after a delay of several days. This type of rupture was only observed following traumatic delivery. Early diagnosis, optically guided orotracheal intubation, and timely treatment can reduce the risks of mortality and morbidity. PMID- 15535141 TI - Management of synchronous subglottic stenosis and bronchial stenosis with a modified endotracheal tube. AB - We present the case of a tracheostomy tube-dependent 6-month-old infant with synchronous airway lesions, including grade III subglottic stenosis and left main bronchomalacia with recurrence of granulation tissue and stenosis. Because the infant was at high risk for complications from the tracheostomy tube, decannulation was the desired outcome. The airway lesions were managed by single stage laryngotracheal reconstruction and short-term stenting of the subglottis and left main bronchus. The greatest challenge was management of progressive stenosis caused by recurrence of granulation tissue in the left main bronchus. A novel modification of a soft silicone elastic endotracheal tube was devised to provide continued ventilation of both lungs while the corrected lesions healed. The role for this type of stent and the benefits and difficulties associated with its use are discussed. PMID- 15535142 TI - Effect of fibrin matrix and vascular endothelial growth factor on reepithelialization of orthotopic murine tracheal transplants. AB - To determine whether the administration of vascular endothelial growth factor (VEGF) alone and in combination with fibrin matrix accelerates murine tracheal allograft reepithelialization, we randomly assigned 40 age-matched mice to 5 experimental groups. BALB/c tracheal grafts were transplanted orthotopically into allogeneic C57BL/6 recipients. The recipients were immunosuppressed with cyclosporine (25 mg/kg per day) and treated with a single topical dose of fibrin matrix, a single topical dose of VEGF, or a single topical dose of a combination of VEGF and fibrin matrix. Thirty-five and 50 days after transplantation, a mixed lymphocyte reaction was performed to assess adequate immunosuppression and the grafts were assessed for rejection, rate and quality of allograft reepithelialization, and cartilage viability. The administration of a combination of fibrin matrix and VEGF to tracheal allografts demonstrated an increased rate of reepithelialization and increased density (37% +/- 2.9%) of morphologically normal ciliated pseudostratified epithelium when compared with an immunosuppressed control group (29.3% +/- 9.1%) 35 days after transplantation. The treated allografts demonstrated no significant change in cartilage viability or rejection. We conclude that the administration of fibrin matrix and VEGF to immunosuppressed tracheal allografts improves the rate and density of tracheal allograft reepithelialization. Carrier-bound growth factors may represent a novel approach to accelerating tracheal allograft reepithelialization and decreasing the need for prolonged immunosuppression following tracheal transplantation. PMID- 15535143 TI - Aberrant recurrent laryngeal nerve reinnervation as a cause of stridor and laryngospasm. AB - Unilateral vocal fold paralysis rarely presents with symptoms of stridor, laryngospasm, and dyspnea. Abnormal reinnervation of abductor nerve branches into adductor fibers may be one cause. Four patients have been positively identified by laryngeal electromyography. Two patients presented after thyroid surgery for thyroid neoplasm. One presented after cervical disk surgery. One patient presented after herpes simplex infection with multiple cranial nerve involvement. All patients had new onset of dyspnea with exertion many months after the recurrent nerve injury. Fiberoptic laryngoscopy showed the affected vocal fold to be immobile. However, with hyperventilation and deep inspiration, there was paradoxical adductor motion of the paretic vocal fold. Laryngeal electromyography showed evidence of reduced but intact voluntary motor units in the thyroarytenoid muscle. These motor units fired on inspiration and not on phonation. This pattern was not seen on the normal side. These findings are consistent with paradoxical innervation and/or synkinesis. Each patient was managed by Botox injection into the adductor muscle. Periodic reinjections may be necessary to manage the condition. PMID- 15535144 TI - Laryngeal electromyographic, cardiovascular, and respiratory effects of neuropeptide injections into the nucleus tractus solitarius of rats. AB - Identifying central neurotransmitters that mediate laryngeal adductor activity may aid in managing pathological laryngeal adduction as occurs in laryngospasm or apparent life-threatening events in infants. We studied the effect on cricothyroid (CT) and thyroarytenoid (TA) electromyography (EMG) and on cardiovascular parameters of neurotransmitter injections into the rat nucleus tractus solitarius (NTS), the primary sensory relay center of the larynx. Twenty nanoliters of vasoactive intestinal peptide (VIP; 2.5 pmol), neurokinin B (NKB, 16 pmol), calcitonin gene-related peptide (3.0 pmol), neurokinin A (NKA; 35 pmol), or artificial cerebrospinal fluid (control solution) was stereotactically injected into the region of the NTS and the control nucleus gracilis in 119 studies performed in 24 mature, anesthetized Sprague-Dawley rats. Changes in diaphragm, CT, and TA EMG activity and blood pressure (BP) were compared. Injection sites were verified histologically. Injections of both VIP and NKB into the region of the NTS, but not the nucleus gracilis, induced life-threatening changes, including apnea, a marked decline in BP (p < .05), and increases in EMG activity of the CT and TA adductor muscles ranging from sustained contraction to mild phasic increases during inspiration. Calcitonin gene-related peptide injection also decreased BP (p < .05), but caused only mild increases in CT and TA EMG activity. NKA and control injections into the NTS did not alter respiration, BP, or CT or TA EMG activity. VIP and NKB may play important roles in modulating EMG activity of the CT and TA muscles. This information may prove useful in evaluating pharmacological targets of central reflex activity to manage life-threatening laryngeal adduction. PMID- 15535145 TI - Cytokeratin-17 as a potential marker for squamous cell carcinoma of the larynx. AB - To assess cytokeratin-17 (CK17) as an immunohistochemical marker for squamous cell carcinoma of the larynx, we stained 63 tissue samples from 63 consecutive patients who were believed or suspected to have squamous cell carcinoma of the larynx for CK17 and analyzed them by computerized histomorphometry. The mean staining intensity for CK17 was significantly stronger (p < .01) in cancerous cells, dysplasia, and normal epithelium proximal to the tumor than in distal normal epithelium and polyps. The percentage of stained area, within samples taken from a single patient, was significantly higher in malignancy and dysplasia as compared to distal normal epithelium and in malignancy as compared to dysplasia and proximal normal epithelium (p < .001). The integrated optical density was significantly higher in the malignant epithelium, dysplasia, polyps, and proximal normal epithelium than in distal normal epithelium (p < .0001). We conclude that CK17 is a highly sensitive and specific immunohistochemical marker for premalignant and malignant transformation in the larynx. Further investigation is warranted in order to assess the role of CK17 in determining safe resection borders. PMID- 15535146 TI - Delayed diagnosis of tracheoesophageal prosthesis aspiration. AB - In 1980, Singer and Blom published the results of their study on use of the tracheoesophageal puncture prosthesis for restoration of voice after total laryngectomy. Since then, the placement of tracheoesophageal puncture prostheses has been an integral part of rehabilitation after laryngectomy. Complications of this procedure have been recognized and are usually minimal. Inadvertent aspiration of the prosthesis is rare. Usually, patients seek help immediately after the incident. We report a case of unnoticed aspiration of a Blom-Singer prosthesis in a patient with a laryngectomy. PMID- 15535147 TI - Role of anaerobic bacteria in infections following tracheostomy, intubation, or the use of ventilatory tubes in children. AB - Colonization of the tracheobronchial tree with microorganisms almost always follows tracheal intubation, tracheostomy, or the use of ventilatory tubes. Infection of the tracheostomy wound site frequently occurs after prolonged use of the tracheostomy. The long-term-ventilated child is at high risk for developing tracheobronchitis or nosocomial pneumonia, generally involving aerobic gram negative or gram-positive bacteria. Several studies have illustrated the role of anaerobic bacteria in lower respiratory tract and tracheostomy wound site infection in intubated children. The predominant anaerobic bacteria were Peptostreptococcus spp and pigmented Prevotella and Porphyromonas spp. Most of these infections are due to polymicrobial aerobic-anaerobic bacterial flora. Appropriate management of mixed pulmonary aerobic and anaerobic infections requires the administration of antimicrobials that are effective against both the aerobic and anaerobic components of the infection. PMID- 15535148 TI - Indications and outcomes of adenoidectomy in infancy. AB - Adenoid hypertrophy is one of the most common causes of pediatric obstructive sleep apnea. Although adenoidectomy is the only effective treatment for adenoid hypertrophy, it is rarely performed in infants less than 1 year old. This study reports on the successful use of adenoidectomy in 24 infants less than 1 year old with a triad of upper airway obstruction symptoms, findings of obstructing adenoids, and obstructive sleep apnea (but no other anomalies). This is a retrospective case series reviewing each infant's clinical data, including presenting symptoms (with special emphasis on apnea episodes), physical examination findings, and results of other investigations such as polysomnography, endoscopy, pH-metry, and echocardiography. With careful preoperative and postoperative monitoring, the 24 infants underwent adenoidectomy without complications. After the procedure, all showed marked improvement with complete disappearance of symptoms of upper airway obstruction, failure to thrive, and gastroesophageal reflux disease. Adenoidectomy was found to be sufficient and curative for such infants. PMID- 15535149 TI - Prevention of hanging columella in open rhinoplasty. AB - In this article we discuss the anatomy of the nasal base and explain the causes of postoperative hanging columella, distinguishing this entity from similar deformities with which it can be confused. We suggest measures that could be helpful in preventing this deformity. We performed external-approach rhinoplasty with the dome suturing method in 82 consecutive patients (64 women and 18 men) from 17 to 44 years of age (mean age, 22 years). Comparative measurements of the alar-columellar complex were made before and after operation. Among the 48 patients with conventional tip suturing (38 women and 10 men), 19 true hanging columellae were detected. In the 34 patients with a modified dome approximation (26 women and 8 men), no more than 5 cases of true deformity were diagnosed. PMID- 15535150 TI - Bezold's abscess following chronic mastoiditis in a newborn. AB - Despite the availability of antibiotics, acute otitis media can still lead to major complications. Bezold's abscesses are very rare complications and are usually found only in adults with well-pneumatized mastoid bones. We present the case of a 10-week-old newborn with a Bezold's abscess. It is stressed that acute otitis media can be silent and undetected in infants and can lead to major complications such as Bezold's abscess. PMID- 15535151 TI - Factors associated with clinical outcomes in acute otitis media. AB - Acute otitis media (AOM) is a common disease in childhood. If predictors of outcome in AOM were known, it would be possible to individualize therapy. Our aim was to identify factors that predict the outcome in AOM. We enrolled 368 children with AOM (ages, 10 to 86 months). The severity of symptoms and the severity of tympanic membrane changes were graded with a scoring system. Nasopharyngeal colonization with middle ear pathogens was determined on day 1. Three outcomes were assessed: persistence of symptoms at day 5, persistence of tympanic membrane changes at day 28, and recurrence of acute symptoms prior to day 28. Persistence of symptoms at day 5 was associated with younger age (35 versus 44 months; p < .001), higher symptom score on day 1 (3.5 versus 2.9; p < .05), and colonization with Streptococcus pneumoniae (61% versus 41%; p < .05). Persistence of tympanic membrane changes at day 28 was associated with younger age (39 versus 45 months; p < .01), higher tympanic membrane score on day 1 (4.1 versus 3.6; p < .01), and nasopharyngeal colonization with S. pneumoniae, especially drug-resistant S. pneumoniae (33% versus 13%; p < .05). Recurrence of acute symptoms prior to day 28 occurred in 14% of the children. Streptococcus pneumoniae was the only pathogen associated with an increased recurrence rate (23%) as compared to the group without pathogens (7%; p < .05). Age, severity of disease at presentation, and nasopharyngeal colonization patterns were proven to be important determinants of outcome in AOM. PMID- 15535152 TI - Formulary restrictions sometimes harm patients. PMID- 15535153 TI - DM industry jumps for joy over Medicare's leap of faith. PMID- 15535154 TI - 'Dr. Jeeves' gives rich medical star treatment. PMID- 15535155 TI - Does payment method drive procedure rates? PMID- 15535156 TI - Not so much of a reach: let sick pay less for drugs. PMID- 15535157 TI - Consumer-directed care bets against human nature. PMID- 15535158 TI - Evaluating new therapies for psoriasis. PMID- 15535160 TI - Plan lets ombudsman go to bat for members. PMID- 15535159 TI - Formulary standards key to E-prescribing success. PMID- 15535161 TI - More plans discover value of health risk assessments. PMID- 15535162 TI - Wound healing promoted with living-skin substitutes. PMID- 15535163 TI - Medical, prescription drug cost rates to drop in 2005. PMID- 15535164 TI - The effects of age on the neural correlates of successful episodic retrieval: an ERP study. AB - The neural correlates of successful episodic retrieval (recollection), as reflected in event-related potentials (ERPs), were investigated in young (ca 20 years; n = 18) and older (ca 70 years; n = 16) healthy individuals. Subjects classified a series of pictures according to whether each item was new or had been encountered at study in the context of an animacy or a size judgment task. By manipulating the number of times items were presented for study, subsets of test items were formed for which source accuracy did not differ according to age. Relative to ERPs elicited by unstudied pictures, ERPs elicited by items attracting equivalent levels of source accuracy showed marked age-related differences. Those from younger subjects demonstrated the positive-going left parietal and right frontal old/new effects described in several previous studies of source memory. By contrast, analogous ERPs from older subjects contained a large left-lateralized negative effect that overshadowed the positive-going effects evident in the young. No age-related differences in either parietal or frontal ERP old/new effects were detected at electrode sites overlying the right hemisphere. It is possible that the age-related ERP differences observed in this task primarily reflect the use of different kinds of information as a basis for source judgments. PMID- 15535165 TI - How arousal modulates memory: disentangling the effects of attention and retention. AB - Emotion may influence memory both by altering attention and perception during encoding and by affecting memory retention. To date, studies have focused on the enhancement of memory consolidation by arousal. However, they have failed to rule out a role for attention. To specifically link memory enhancement of arousing material to modulation of memory retention, we examined recognition of neutral and arousing words at two time points and under conditions that manipulate attention during encoding. Participants were briefly presented with an arousing or neutral word at the periphery, while fixating on a central word. Recognition of peripheral words was assessed either immediately or after 24 h. Whereas recognition of neutral words became worse over time, recognition of arousing words remained the same and was better than neutral word recognition at delay. The results indicate that arousal supports slower forgetting even when the difference in attentional resources allocated to stimuli is minimized. PMID- 15535166 TI - The impact of orbital prefrontal cortex damage on emotional activation to unanticipated and anticipated acoustic startle stimuli. AB - Damage to the orbital prefrontal cortex has been implicated in selectively diminishing electrodermal autonomic nervous system responses to anticipated punishing stimuli (e.g., losing money; Bechara, Damasio, & Damasio, 2000), but not to unanticipated punishing stimuli (e.g., loud noises; Damasio, Tranel, & Damasio, 1990). We extended this research by examining the effects of orbitofrontal damage on emotional responses to unanticipated and anticipated acoustic startles and collecting a more extensive set of physiological measures, emotional facial behavior, and self-reported emotional experience. Consistent with previous research, patients showed intact physiology to an unanticipated startle but failed to show appropriate anticipatory cardiovascular responses (patients' heart rates decreased, controls' increased). In addition, patients displayed more surprise facial behavior and reported marginally more fear than did controls in response to the unanticipated startle. Thus, orbitofrontal damage may compromise the ability to anticipate physiologically the onset of aversive stimuli, despite intact or enhanced emotional responses when such stimuli occur unexpectedly. PMID- 15535167 TI - Amygdala and hippocampal activity during acquisition and extinction of human fear conditioning. AB - Previous functional magnetic resonance imaging (fMRI) studies have characterized brain systems involved in conditional response acquisition during Pavlovian fear conditioning. However, the functional neuroanatomy underlying the extinction of human conditional fear remains largely undetermined. The present study used fMRI to examine brain activity during acquisition and extinction of fear conditioning. During the acquisition phase, participants were either exposed to light (CS) presentations that signaled a brief electrical stimulation (paired group) or received light presentations that did not serve as a warning signal (control group). During the extinction phase, half of the paired group subjects continued to receive the same treatment, whereas the remainder received light alone. Control subjects also received light alone during the extinction phase. Changes in metabolic activity within the amygdala and hippocampus support the involvement of these regions in each of the procedural phases of fear conditioning. Hippocampal activity developed during acquisition of the fear response. Amygdala activity increased whenever experimental contingencies were altered, suggesting that this region is involved in processing changes in environmental relationships. The present data show learning-related amygdala and hippocampal activity during human Pavlovian fear conditioning and suggest that the amygdala is particularly important for forming new associations as relationships between stimuli change. PMID- 15535168 TI - Enhanced occipital and anterior cingulate activation in men but not in women during exposure to angry and fearful male faces. AB - Blood-oxygenation-level-dependent signal was measured with functional magnetic resonance imaging in 24 healthy young subjects (12 men and 12 women) during viewing of angry, fearful, and neutral male and female face pictures. Exposure to angry male as opposed to angry female faces activated the visual cortex and the anterior cingulate gyrus significantly more in men than in women. A similar sex differential brain activation pattern was present during exposure to fearful but not neutral faces. Previous behavioral studies indicate enhanced physiological arousal in men but not in women during exposure to angry male asopposed to female faces, and brain imaging studies have shown that the occipital cortex and the anterior cingulate gyrus are influenced by activity in the autonomic nervous system as well as by visualattention. Hence, we suggest that the elevated occipital and anterior cingulate activation in men during confrontation with other angry and fearful males may reflect enhanced vigilance in a potentially dangerous situation. PMID- 15535169 TI - Not all synaesthetes are created equal: projector versus associator synaesthetes. AB - In synaesthesia, ordinary stimuli elicit extraordinary experiences. When grapheme color synaesthetes view black text, each grapheme elicits a photism-a highly specific experience of color. Importantly, some synaesthetes (projectors) report experiencing their photisms in external space, whereas other synaesthetes (associators) report experiencing their photisms "in the mind's eye." We showed that projectors and associators can be differentiated not only by their subjective reports, but also by their performance on Stroop tasks. Digits were presented in colors that were either congruent or incongruent with the synaesthetes' photisms. The synaesthetes named either the video colors of the digits or the colors of the photisms elicited by the digits. The results revealed systematic differences in the patterns of Stroop interference between projectors and associators. Converging evidence from first-person reports and third-person objective measures of Stroop interference establish the projector/ associator distinction as an important individual difference in grapheme-color synaesthesia. PMID- 15535170 TI - Why aren't all deep superior colliculus neurons multisensory? A Bayes' ratio analysis. AB - Multisensory neurons in the deep superior colliculus (SC) show response enhancement to cross-modal stimuli that coincide in time and space. However, multisensory SC neurons respond to unimodal input as well. It is thus legitimate to ask why not all deep SC neurons are multisensory or, at least, develop multisensory behavior during an organism's maturation. The novel answer given here derives from a signal detection theory perspective. A Bayes' ratio model of multisensory enhancement is suggested. It holds that deep SC neurons operate under the Bayes' ratio rule, which guarantees optimal performance-that is, it maximizes the probability of target detection while minimizing the false alarm rate. It is shown that optimal performance of multisensory neurons vis-a-vis cross-modal stimuli implies, at the same time, that modality-specific neurons will outperform multisensory neurons in processing unimodal targets. Thus, only the existence of both multisensory and modality-specific neurons allows optimal performance when targets of one or several modalities may occur. PMID- 15535171 TI - Is detecting prospective cues the same as selecting targets? An ERP study. AB - Prospective memory represents our ability to realize intentions that must be delayed for some period of time. In this study, we examined modulations of the event-related brain potentials (ERPs) associated with target selection in visual working memory and prospective-cue detection in prospective memory. Targets and prospective cues elicited an N2pc, indicating that a common neural mechanism supports selection in working memory and prospective memory. Partial least squares analysis revealed that the N300 and prospective positivity were associated with a latent variable that contrasted the ERPs elicited by prospective-cue trials with those elicited by target-present and target-absent trials, in agreement with the idea that these modulations of the ERPs are uniquely related to prospective memory. PMID- 15535172 TI - Cortical regions associated with different aspects of object recognition performance. AB - In the present object recognition study, we examined the relationship between brain activation and four behavioral measures: error rate, reaction time, observer sensitivity, and response bias. Subjects perceptually matched object pairs in which structural similarity (SS), an index of structural differentiation, and exposure duration (DUR), an index of task difficulty, were manipulated. The SS manipulation affected the fMRI signal in the left anterior fusiform and parietal cortices, which in turn reflected a bias to respond same. Conversely, an SS-modulated fMRI signal in the right middle frontal gyrus reflected a bias to respond different. The DUR manipulation affected the fMRI signal in occipital and posterior fusiform regions, which in turn reflected greater sensitivity, longer reaction times, and greater accuracy. These findings demonstrate that the regions most strongly implicated in processing object shape (SS-modulated regions) are associated with response bias, whereas regions that are not directly involved in shape processing are associated with successful recognition performance. PMID- 15535173 TI - The neural bases of the effects of item-nonspecific proactive interference in working memory. AB - We reanalyzed the behavioral and fMRI data from seven previously published studies of working memory in order to assess the behavioral and neural effects of item-nonspecific proactive interference (PI; attributable to the accrual of antecedent information independent of the repetition of particular items). We hypothesized that item-nonspecific PI, implicated in age-related declines in working memory performance, is mediated by the same mechanism(s) that mediate item-specific PI (occurring when an invalid memory probe matches a memorandum from the previous trial). Reaction time increased across trials as a function of position within the block, a trend that reversed across the duration of each multiblock experiment. The fMRI analyses revealed sensitivity to item-nonspecific PI during the probe epoch in the left anterior inferior frontal gyrus and the left dorsolateral prefrontal cortex(PFC). They also revealed a negative trend, across trials, in the transient probe-evoked component of the global signal. A common PFC-based mechanism may mediate many forms of PI. PMID- 15535175 TI - Cerebrospinal fluid dynamics. AB - Hydrocephalus is far more complicated than a simple disorder of CSF circulation. Historically, it has been diagnosed using clinical and psychomotor assessment plus brain imaging. The role of physiological measurement to aid diagnosis becomes more appreciated in current clinical practice. This has been reflected by recently formulated guidelines for the management of normal pressure hydrocephalus. Clinical measurement in hydrocephalus is mainly related to intracranial pressure (ICP) and cerebral blood flow. This review lists and discusses most common forms of the methods: CSF infusion study, overnight ICP monitoring, assessment of slow ICP waves, testing pressure reactivity, cerebral autoregulation, CO2 reactivity and PET-CBF studies combined with MRI co registration. The basics of CSF dynamics modelling are presented and the principles of the assessment of functioning of the implanted hydrocephalus shunts are also discussed. The descriptions of multiple forms of measurement along with clinical illustrations are mainly based on in-house experience of a multidisciplinary group of scientists and clinicians from Cambridge, UK. PMID- 15535174 TI - Mild hypoxia disrupts recollection, not familiarity. AB - Yonelinas et al. (2002) found that hypoxic patients exhibited deficits in recollection that left familiarity relatively unaffected. In contrast, Manns, Hopkins, Reed, Kitchener, and Squire (2003) studied a group of hypoxic patients who suffered severe and equivalent deficits in recollection and familiarity. We reexamine those studies and argue that the discrepancy in results is likely due to differences in the hypoxic groups that were tested (i.e., differences in amnestic severity, subject sampling methods, and patient etiology). Yonelinas et al. examined memory in 56 cardiac arrest patients who suffered a brief hypoxic event, whereas Manns et al. examined a group of severely amnesic patients that consisted of 2 cardiac arrest patients, 2 heroin overdose patients, 1 carbon monoxide poisoning patient, and 2 patients with unknown etiologies. We also consider an alternative explanation proposed by Wixted and Squire (2004), who argued that the two patient groups suffered similar deficits, but that statistical or methodological artifacts distorted the results of each of Yonelinas et al.'s experiments. A consideration of those results, however, indicates that such an explanation does not account for the existing data. All of the existing evidence indicates that recollection, but not familiarity, is disrupted in mild hypoxic patients. In more severe cases of hypoxia, or those with more complex etiologies such as heroin overdose, more profound deficits may be observed. PMID- 15535176 TI - Effect of electrode locations and respiration in the characterization of myocardial tissue using a transcatheter impedance method. AB - Our objective is to evaluate whether it is possible to characterize the passive electrical properties of myocardial tissue in contact with the electrocatheters used in arrhythmia diagnosis or radio frequency ablation techniques. To characterize the tissue, we propose the use of electrical impedance spectroscopy to measure the impedance between the catheter tip and an external electrode, assuming a three-electrode method. We constructed a 3D finite-element model of the thorax to estimate the impedance as measured in different situations. We defined an area on the anterior wall of the left ventricle in which we simulated three tissue states: healthy, acute ischaemic and scar. We studied the effect of the following parameters on the measured impedance spectrum: the position of the external electrode, the position and orientation of the catheter tip and the overall effect of the subject's respiration. Results show that the highest frequency phase (around 300 kHz) yields the best differentiation of tissue states and that it is less sensitive to respiration than the impedance magnitude. The phase is also less influenced by the catheter tip position (either touching the wall or floating) and the orientation of the catheter inside the left ventricle. The best position for the external electrode is on the chest; this position is less affected by breathing and is more sensitive to tissue changes. One can still distinguish between tissue states if the external electrode is placed on the back, but the effect of respiration is higher. PMID- 15535177 TI - Instability and frequency-domain variability of heart rates in fetuses with or without growth restriction affected by severe preeclampsia. AB - This study investigated how the instability and frequency-domain variability in heart rates differ between fetuses affected only by severe preeclampsia and fetuses affected by both severe preeclampsia and growth restriction. From their antepartum fetal heart rates and those of control fetuses, the very short-term intermittency (C1alpha) and the spectral powers were calculated to evaluate the instability and frequency-domain variability, respectively. The fetuses affected only by severe preeclampsia showed abnormally high C1alpha and low- and high frequency power. The fetuses affected by severe preeclampsia and growth restriction showed even higher C1alpha than that of the fetuses affected by severe preeclampsia and abnormally reduced low-frequency power. Conclusively, when compared to the heart rates of fetuses affected only by severe preeclampsia, the heart rates of fetuses affected by severe preeclampsia and growth restriction showed a greater abnormal instability and an abnormally reduced variability at low-frequency range. PMID- 15535178 TI - Agreement between three commercially available instruments for measuring short term heart rate variability. AB - Numerous instruments are commercially available to measure heart rate variability, yet little is known regarding the agreement between such instruments. The objective of this study is to assess agreement between measures of heart rate variability in three commercially available instruments. Thirty subjects (20 males) of median age 27.5 (range 19-59 years) underwent simultaneous ECG recordings, under three different resting conditions: supine, standing and supine with controlled breathing, using three commercially available analysers. Intraclass correlation coefficients tended to show excellent agreement (lower 95% C.I., R > 0.75) between all instruments under all conditions. However, further analysis of selected measurements using the limits of agreement method revealed large variation in values generated by all instruments. There was also an evidence of systematic bias between one instrument and the remaining two. The latter finding was due to discrepant ECG recording protocols that were unrelated to consistent operator timing. This study demonstrates that measures of HRV generated by the three instruments did not agree well in all cases. Discrepancies were due to the recording protocols of the systems. This may lead to incomparable results between instruments. It is therefore recommended that: (a) if different instruments are used in the same study or (b) multi-centre study designs are planned or (c) heart rate variability results are discussed with reference to studies using other instruments, levels of agreement need to be reported to ensure comparability. PMID- 15535179 TI - Validity of measurements for cycle-by-cycle variability of jaw movements: variability of chewing cycles in cases of prognathism. AB - Objective assessments of masticatory jaw movements for patients with a high degree of occlusal abnormalities are critical to elucidate the relationship between occlusal function and morphology. This study aimed to test the reliability and validity of measuring specific cycle-by-cycle variability of chewing jaw movements of patients with mandibular prognathism. The lower incisor point movements were monitored using a 3D tracking device. Analyses consist of evaluations for the planarity of each cycle, cycle-by-cycle variances in orientations of the best-fit planes and instantaneous curvature of the trajectories. These spatio-temporal variabilities of chewing cycles for 12 adult females with good occlusion were compared with those for 12 female adult patients with mandibular prognathism. Five of the control subjects performed chewing on two separate occasions. Measurement outcomes were repeatable within each subject. For both groups, each single chewing cycle was characterized as reasonably planar. Orientations of the best-fit planes and curvatures of the trajectories during closing were more variable for patients than those for the control subjects. These findings suggest certain effectiveness of the present measurements of orientations of the best-fit planes and movement curvatures for scaling the variable nature of patients' masticatory jaw movements. PMID- 15535180 TI - Comprehensive analysis of cardiac health using heart rate signals. AB - The electrocardiogram is a representative signal containing information about the condition of the heart. The shape and size of the P-QRS-T wave, the time intervals between its various peaks, etc may contain useful information about the nature of disease affecting the heart. However, the human observer cannot directly monitor these subtle details. Besides, since bio-signals are highly subjective, the symptoms may appear at random in the time scale. Therefore, the heart rate variability signal parameters, extracted and analyzed using computers, are highly useful in diagnostics. Analysis of heart rate variability (HRV) has become a popular noninvasive tool for assessing the activities of the autonomic nervous system. The HRV analysis is based on the concept that fast fluctuations may specifically reflect changes of sympathetic and vagal activity. It shows that the structure generating the signal is not simply linear, but also involves nonlinear contributions. These signals are essentially nonstationary; may contain indicators of current disease, or even warnings about impending diseases. The indicators may be present at all times or may occur at random in the time scale. However, to study and pinpoint abnormalities in voluminous data collected over several hours is strenuous and time consuming. This paper deals with the analysis of eight types of cardiac abnormalities and presents the ranges of linear and nonlinear parameters calculated for them with a confidence level of more than 90%. PMID- 15535181 TI - Filtering artefacts in measurements of forced oscillation respiratory impedance in young children. AB - Respiratory mechanical impedance measured at a single frequency is of great interest to describe change in airways dimensions with time in young children with asthma. Adequate filtering is mandatory in order to eliminate spurious values at rapid flow transient or during glottis closure. The aim of the study was to test a new filtering procedure. Respiratory impedance was measured at 12 Hz using the head generator technique in 33 asthmatic children aged 3.5-7.5 years. Two separate measurements were obtained at baseline and one 10 min after salbutamol inhalation. Assuming corrupted data would correspond to major distortion in flow oscillation, the tested procedure calculated an index expressing departure of each flow oscillation from an ideal sinusoid (flow shape index filter (FSIF)). Cut-offs ranging from 5 to 20% were compared to the visual identification and 'by hand' elimination of corrupted data (visual filtering). Sensitivity and specificity of FSIF versus visual filtering indicated an optimal range of cut-off values between 10 and 15% (FSIF10 and FSIF15). Compared with visual filtering, respiratory conductance in inspiration (Grs) was marginally but significantly lower with FSIF10 (p < 0.01) but not FSIF15 and had slightly larger (within measurement) coefficient of variation with either FSIF10 or FSIF15 (p < 0.01). Between-measurement repeatability of Grs and Grs change induced by salbutamol were similarly estimated by FSIF10, FSIF15 or visual filtering. It is concluded that FSIF appears as a valid alternative to visual filtering. The 10% cut-off is a good compromise considering sensitivity (72%), amount of rejected data (<20%) and effect on Grs and variability. PMID- 15535182 TI - Real time detection of ventricular fibrillation and tachycardia. AB - The automatic external defibrillator (AED) is a lifesaving device, which processes and analyses the electrocardiogram (ECG) and delivers a defibrillation shock to terminate ventricular fibrillation or tachycardia above 180 bpm. The built-in algorithm for ECG analysis has to discriminate between shockable and non shockable rhythms and its accuracy, represented by sensitivity and specificity, is aimed at approaching the maximum values of 100%. An algorithm for VF/VT detection is proposed using a band-pass digital filter with integer coefficients, which is very simple to implement in real-time operation. A branch for wave detection is activated for heart rate measurement and an auxiliary parameter calculation. The method was tested with ECG records from the widely recognized databases of the American Heart Association (AHA) and the Massachusetts Institute of Technology (MIT). A sensitivity of 95.93% and a specificity of 94.38% were obtained. PMID- 15535183 TI - Defibrillation shock success estimation by a set of six parameters derived from the electrocardiogram. AB - It is well known that in some cases defibrillator shocks cannot terminate ventricular fibrillation (VF). Repeated failed shocks often may worsen subsequent response to therapy. This study assesses the ability of six parameters derived from the surface electrocardiogram (ECG) to predict defibrillation shock outcome. Using stepwise discriminant analysis, we obtained several discriminant functions, yielding different combinations of sensitivity and specificity for detection of pre-shock ECG segments corresponding to successful versus unsuccessful shocks. The study was performed consecutively for 3, 4 and 5 s ECG time intervals. The prediction accuracy of 72.3% (61.8% sensitivity and 79.6% specificity) with five parameters and 3 s VF segment analysis prior to defibrillation shock could be considered acceptable for possible practical application in automatic external defibrillators. PMID- 15535184 TI - Measurement reliability of highly variable physiological responses to experimentally-manipulated gas fractions. AB - Ventilatory and cardiac responses to changing inhaled gas fractions are notoriously variable within individuals. Such variation can confound clinical diagnoses and hypotheses about human adaptation. In this study we use a cardiac (HHR) and a ventilatory (HVR) measure of physiological sensitivity to an experimentally manipulated oxygen concentration (8% O2), to compare variation (a) within and between individuals, (b) within and between days and (c) within and between physiological parameters. To explore the sources of variation, we use the coefficient of variation (CV, %), repeatability (R, intraclass correlation coefficient, %) and repeated-measures analyses of variance. Both the HVR and the HHR are significantly repeatable (HVR: R = 0.76-0.92; HHR: R = 0.35-0.76) and equally variable within and between days. Its high R suggests that the HVR displays greater between-individual variation relative to within-individual variation than does the HHR. The HVR is thus a more reliable measure of physiological sensitivity to hypoxia than is the HHR. We suggest how these results may inform experimental design, and suggest how to avoid stochastic and experimental artefacts when investigating ventilatory and cardiac physiological responses to hypoxia. PMID- 15535185 TI - Modulated photoacoustic spectroscopy study of an artificial tanning on human skin induced by dihydroxyacetone. AB - A modulated photoacoustic spectroscopy study on the effect of dihydroxyacetone, commonly used for artificial tan, is presented. The study was carried out in the presence and absence of dimethylisosorbide (a solvent for dihydroxyacetone) on fresh human skin, obtained from the breast region of recent autopsy cases (two females), at a frequency of 25 Hz, which enabled us to study the effect at a depth of 30 microm in the stratum corneum and beneath. By monitoring the photoacoustic signal intensity with time in the region of 300-400 nm, which is a specific region for melanin pigment, it is demonstrated that dihydroxyacetone in combination with dimethylisosorbide enhances the process of tanning. Dihydroxyacetone also has an effect on the amino acids and nucleic acids which is bad for the skin. PMID- 15535186 TI - Torque properties of a rat oesophagus for physiological and diabetic conditions. AB - In this paper the torque of an oesophagus is studied for physiological and diabetic conditions. Since the function of the oesophagus is mainly mechanical, this work is focused on providing quantitative measurement of the passive biomechanical properties of the oesophagus torque. The oesophagus was treated as a membrane when calculating the stress and strain. The torque versus twist-angle relation was approximated to be linear at a specified pressure and longitudinal stretch ratio. Thus, the shear modulus can be computed by the torque, twist angle and polar moment of inertia in this state. The shear modulus varies greatly with the changing inflation pressure and longitudinal stretch ratio. When the longitudinal stretch ratio or transmural pressure is constant, the shear modulus is increased after 28 days of diabetes. PMID- 15535187 TI - A novel implantable cardiac telemetry system for studying atrial fibrillation. AB - Atrial fibrillation (AF) is the most common arrhythmia in clinical practice. Most in vivo experimental research on AF is performed in a surgical setting, on animals instrumented by external devices, or using commercial implantable pacemakers. This paper describes a novel implantable cardiac telemetry system, which allows the study of AF remotely in conscious and ambulatory animals over a few month period. To validate this concept, the system was built and implanted in a sheep for 3 months. During this period, the system was used to deliver chronic rapid atrial pacing for AF induction, and to record and measure atrial electrograms and atrial effective refractory period (AERP) daily. During the course of AF induction the AERP decreased, confirming the progression of the electrical remodeling process in the atria. Episodes of paroxysmal AF were successfully induced in the animal. Burst pacing therapy was delivered with the system, however, no AF termination was observed. Result shows that this telemetry based pacing and monitoring system can be used to study AF in a conscious animal non-invasively for an extended period of time, making this system a unique research tool. PMID- 15535188 TI - Bilateral comparison of blackbody cavities for calibration of infrared ear thermometers between NPL and FE/LMK. AB - The paper describes the comparison method and analyses the results of comparison in terms of agreement between the blackbody of the National Physical Laboratory (NPL), United Kingdom and four different blackbody cavities of the Laboratory of Metrology and Quality (LMK), at the University of Ljubljana, Faculty of Electrical Engineering (FE), Slovenia. Three cavity shapes are suggested in different standards as suitable for calibration of infrared ear thermometers (IRETs), while one cavity shape was proposed by the LMK. The agreement between blackbody cavities was determined with the help of platinum resistance thermometers. Two reference IRETs were used to check their stability and level of agreement between calibration results at different institutes and against different blackbody cavities. Measurements were performed with two IRETs, at the NPL in one cavity and at the LMK in four different cavities. The comparison was initiated to solve the problem of assuring proper traceability for IRETs and to present the solution to their users. PMID- 15535189 TI - A probe for immittance spectroscopy based on the parallel electrode technique. AB - This paper presents the construction of a probe for immittance spectroscopy. The probe is based on the four-electrode technique. The current application electrodes are long parallel strips and have been partitioned into three segments to measure the immittance of relatively small volumes. The probe constant has been calculated assuming anisotropy of the materials examined. The experiments, based on the measurements of KCl solutions and numerical simulations (finite element method--FEM), have shown a high degree of consistency with the theoretical value. Experimental in vivo measurements on swine gluteal tissue were performed and the conductivity and permittivity reconstructed from these measurements are presented. PMID- 15535190 TI - Gastric activity studies using a magnetic tracer. AB - A magnetic pulse generator has been set up in order to study gastric activity. Two coils 1.05 m in diameter, arranged in a Helmholtz configuration, were used. The system generated magnetic field pulses higher than 15 mT, of duration 17.3+/ 1.2 ms. Measurements were performed in 11 male volunteers, with average age 29.3+/-6.4 years and body mass index 26.0+/-4.8 kg m(-2). Magnetite (Fe3O4) particles with diameters from 75 to 125 microm were used as magnetic tracers, which were mixed in 250 ml of yogurt in concentrations from 2 to 5 g. Signals were registered by using a high speed 3 axis fluxgate digital magnetometer and processed to determine the relaxation of the magnetic tracers by fitting a first order exponential function to the data, a mean relaxation constant K = 116+/-40 s(-1) was obtained. Also, an average gastric peristaltic frequency was measured; a value of 3.2+/-0.3 cpm was determined. PMID- 15535191 TI - Bioelectrical impedance vector analysis in haemodialysis patients: relation between oedema and mortality. AB - In this work, bioelectrical impedance vector analysis (BIVA) method is used in a sample of haemodialysis patients in stable (without oedema) and critical (hyperhydrated and malnutrition) states, in order to establish the relation between hyperhydration (oedema) and mortality. The measurements obtained were single frequency (50 kHz), tetrapolar (hand-foot) complex impedance measurements (vector components are: resistance R and reactance Xc). The impedance components were standardized by the height H of the subjects, (R/H and Xc/H) to obtain de impedance vector Z/H, that is represented in the RXc plot (abscise R/H, ordinate Xc/H). Measurements were performed on a sample of 74 patients (30 men and 44 women, 18-70 year, body mass index (BMI), 19-30 kg m(-2)) at the Saturnino Lora University Hospital in Santiago de Cuba. The 46 stable patients comprised 28 men and 18 women; the 28 critical patients 16 men and 12 women. The reference population consisted of 1196 healthy adult subjects living in Santiago de Cuba (689 men and 507 women, 18-70 year, BMI 19-30 kg m(-2)). We used the RXc plot with the BIVA method to characterize the reference population using the 50%, 75% and 95% tolerance ellipses. Student's t-test and Hotelling's T2-test were used to analyse the separation of groups obtained by means of clinical diagnosis and those obtained by BIVA. We obtained a significant difference (P < 0.05) in R/H, Xc/H and phase angle (PA) in men as in women between the location of Z/H vectors in the RXc graph and the separation made by the doctors between stable and critical patients. Critical (hyperhydrated) patients were located below the inferior pole of the 75% tolerance ellipse, whereas stable patients were within the tolerance ellipses. Some cases classified as stable by the clinic were classified as hyperhydrated by BIVA with 100% sensitivity and 48% specificity. In conclusion, the BIVA method could be used to classify patients by hydration state and to predict survival. Advantages of the method are its simplicity, objectivity and that it does not require the definition of patient dry weight. PMID- 15535192 TI - Ranking of pattern recognition parameters for premature ventricular contractions classification by neural networks. AB - Detection and classification of ventricular complexes from a limited number of ECG leads is of considerable importance in critical care or operating room patient monitoring. Beat-to-beat detection allows the heart rhythm evolution to be followed and various types of arrhythmia to be recognized. A quantitative analysis is proposed of pattern recognition parameters for classification of normal QRS complexes and premature ventricular contractions (PVC). Twenty-six parameters have been defined: the width of the QRS complex, three vectorcardiogram parameters and 11 from two ECG leads. These parameters include: amplitudes of positive and negative peaks, area of positive and negative waves, various time-interval durations, amplitude and angle of the QRS vector, etc. They are measured for all QRS complexes annotated as 'normals' and 'PVCs' from the 48 ECG recordings of the MIT-BIH arrhythmia database. Neural networks (NN) are shown to be a useful instrument for the analysis of large quantities of parameters. Separate ranking of any parameter and homogeneous group ranking (amplitude, area, interval, slope and vector) were performed. From the two ECG leads, the first three ranked parameter groups for clustering of PVCs are amplitude, slope and interval, while for N clustering they are vector, amplitude and area. Considering the entire parameter set, we obtained N = 99.7% correct detection of normal QRS complexes and PVC = 98.5% of premature ventricular complexes. The study also shows that simultaneous analysis of two ECG channels yields better accuracy compared to using a single channel: the improvement is 0.1% in the classification of N beats and 4.5% for PVC beats. PMID- 15535193 TI - Activity-based sleep-wake identification in infants. AB - Actigraphy offers one of the best-known alternatives to polysomnography for sleep wake identification. The advantages of actigraphy include high accuracy, simplicity of use and low intrusiveness. These features allow the use of actigraphy for determining sleep-wake states in such highly sensitive groups as infants. This study utilizes a motion sensor (accelerometer) for a dual purpose: to determine an infant's position in the crib and to identify sleep-wake states. The accelerometer was positioned over the sacral region on the infant's diaper, unlike commonly used attachment to an ankle. Opposed to broadly used discriminant analysis, this study utilized logistic regression and neural networks as predictors. The accuracy of predicted sleep-wake states was established in comparison to the sleep-wake states recorded by technicians in a polysomnograph study. Both statistical and neural predictors of this study provide an accuracy of approximately 77-92% which is comparable to similar studies achieving prediction rates of 85-95%, thus validating the suggested methodology. The results support the use of body motion as a simple and reliable method for determining sleep-wake states in infants. Nonlinear mapping capabilities of the neural network benefit the accuracy of sleep-wake state identification. Utilization of the accelerometer for the dual purpose allows us to minimize intrusiveness of home infant monitors. PMID- 15535194 TI - Time course reconstruction of fetal cardiac signals from fMCG: independent component analysis versus adaptive maternal beat subtraction. AB - M-mode and pulsed Doppler echocardiography, cardiotocography and transabdominal fetal ECG are available in clinical practice to monitor fetal cardiac activity during advancing gestation, but none of these methods allows the direct measurement of morphological and temporal parameters for fetal rhythm assessment. Fetal magnetocardiograms (fMCGs) are noninvasive recordings of magnetic field variations associated with electrical activity of the fetal heart obtained with superconducting sensors positioned over the maternal abdomen inside a shielded room. Because of maternal cardiac activity, fMCGs are contaminated by maternal components that need to be eliminated to reconstruct fetal cardiac traces. The aim of the present work was to use two methods working in the time domain, an independent component analysis algorithm (FastICA) and an adaptive maternal beat subtraction technique (AMBS), for the retrieval of fetal cardiac signals from fMCGs. Detection rates of both methods were calculated, and FastICA and AMBS performances were compared in the context of clinical applications by estimating several temporal and morphological characteristics of the retrieved fetal traces, such as the shape and duration P-QRS-T waves, arrhythmic beat detection and classification, and noise reduction. Quantitative and qualitative comparison produced figures that always suggested that FastICA was superior to AMBS from the perspective of clinical use of the recovered fetal signals. PMID- 15535195 TI - The effect of contacting force on photoplethysmographic signals. AB - Photoplethysmography (PPG) has been used in oxygen saturation measurement, heart rate monitoring, and the assessment of peripheral circulation and large artery compliance. However, the waveform of the photoplethysmographic signal may be affected by the contacting force between the sensor and the measurement site. The aim of this study is to investigate the change in pulse amplitude (AC), DC amplitude, ratio of AC/DC and normalized pulse area of the reflective photoplethysmographic signals with increasing contacting force, from 0.2 N to 1.8 N. Signals were recorded from the fingers of fifteen healthy subjects. With increasing contacting force, the DC amplitude increased and the normalized pulse area decreased, whereas the pulse amplitude and the ratio of AC/DC increased first and then decreased. For different subjects, the pulse amplitude and the ratio of AC/DC peaked at different contacting forces, from 0.2 N to 1.0 N, and most of the subjects achieved their maximum pulse amplitude within 0.2-0.4 N. Over the range of contacting force between 0.2 N and 0.8 N, the DC amplitude and the normalized pulse area had significant changes (p < 0.001). The results suggest that the effects of contacting force should be carefully examined in the design of photoplethysmography-based health care devices. PMID- 15535196 TI - [Recommendations for basal cell carcinoma: beware of aggressive types!]. PMID- 15535197 TI - [Recommendations for basal cell carcinoma]. PMID- 15535198 TI - [Cancer of the female genital tract--natural history, epidemiology, and biostatistics]. PMID- 15535199 TI - [Cancer prevention]. PMID- 15535200 TI - [History of cancer therapy]. PMID- 15535201 TI - [Diagnosis and treatment of gynecological cancers]. PMID- 15535202 TI - [History and practice of gynecological cancer registration]. PMID- 15535203 TI - [Registration of gynecologic malignancy in Japan]. PMID- 15535204 TI - [Survival of gynecological cancer patients in Osaka, Japan]. PMID- 15535205 TI - [Epidemiology of uterine cervix cancer]. PMID- 15535206 TI - [Molecular carcinogenesis of the uterine cervix]. PMID- 15535207 TI - [Genetic abnormality in cervical cancer]. PMID- 15535208 TI - [Abnormal fragile histidine triad and cervical cancer]. PMID- 15535209 TI - [Molecular mechanism in carcinogenesis of the uterine cervix]. PMID- 15535210 TI - [Mechanism of metastasis in cervical cancer]. PMID- 15535211 TI - [Pathology of cervical cancer]. PMID- 15535212 TI - [Management of dysplasia of the uterine cervix]. PMID- 15535213 TI - [General rules for clinical and pathological management of uterine cervical cancer]. PMID- 15535214 TI - [Clinical practice guidelines according to clinical stage of cervical cancer]. PMID- 15535215 TI - [Genetic diagnosis of cervical cancer]. PMID- 15535216 TI - [Tumor markers for cancer of the uterine cervix]. PMID- 15535217 TI - [Diagnosis of cervical neoplasm--cytology]. PMID- 15535218 TI - [Topics in histopathological diagnosis of cancer of the uterine cervix]. PMID- 15535219 TI - [Diagnostic imaging of uterine cervical carcinoma]. PMID- 15535220 TI - [Treatment for cervical cancer]. PMID- 15535221 TI - [Surgical treatment for cervical cancer]. PMID- 15535222 TI - [Radical hysterectomy]. PMID- 15535224 TI - [Loop electrosurgical excision procedure]. PMID- 15535223 TI - [Preservation of organic function in surgery for cervical cancer]. PMID- 15535225 TI - [Laser conization]. PMID- 15535226 TI - [Photodynamic therapy for early cervical cancer]. PMID- 15535227 TI - [Chemotherapy for high-risk cervical carcinoma]. PMID- 15535229 TI - [Taxol in advanced cervical cancer]. PMID- 15535228 TI - [Irinotecan hydrochloride (CPT-11) for advanced cervical cancer]. PMID- 15535230 TI - [Chemotherapy using nedaplatin for advanced gynecologic cancers]. PMID- 15535231 TI - [BOMP (BLM, VCR, MMC, and CDDP) therapy for advanced cervical cancer]. PMID- 15535232 TI - [Neoadjuvant chemotherapy for cervix neoplasms]. PMID- 15535233 TI - [Intraarterial chemotherapy for cervix neoplasms]. PMID- 15535234 TI - [Chemotherapy with mitomycin C, etoposide and cisplatin for adenocarcinoma of the uterine cervix]. PMID- 15535235 TI - [Response evaluation criteria in chemotherapy for cervix neoplasm]. PMID- 15535236 TI - [Radiotherapy for cervical cancer]. PMID- 15535237 TI - [Para-aortic lymphadenectomy for cervix cancer]. PMID- 15535238 TI - [Lymphadenectomy for cervical cancer: indications and limits]. PMID- 15535239 TI - [Hemoglobin level and recurrent rate of uterine cervical cancer]. PMID- 15535240 TI - [Treatment of cervical carcinoma. Prognostic factors in patients with surgical treatment]. PMID- 15535241 TI - [Recurrence of uterine cervical cancer more than 5 years after initial therapy]. PMID- 15535243 TI - [Current status, problems and answers in cervical cancer screening]. PMID- 15535242 TI - [Risk factors and prevention of uterine cervical cancer]. PMID- 15535244 TI - [Epidemiology of endometrial carcinoma]. PMID- 15535245 TI - [Molecular mechanisms in endometrial carcinoma]. PMID- 15535246 TI - [Genetic abnormalities in endometrial cancer]. PMID- 15535247 TI - [Topics on gene abnormality in endometrial cancer]. PMID- 15535248 TI - [Biology of endometrial cancer]. PMID- 15535249 TI - [Metastasizing mechanisms in endometrial cancer]. PMID- 15535250 TI - [Pathology of endometrial carcinoma]. PMID- 15535252 TI - [General rules for clinical and pathological management of uterine corpus cancer]. PMID- 15535251 TI - [Borderline lesions in the endometrium]. PMID- 15535253 TI - [Stage classification and clinical practice guidelines in endormetrial carcinomas]. PMID- 15535254 TI - [Genetic diagnosis of endometrial cancer]. PMID- 15535255 TI - [Tumor markers for carcinoma of the endometrium]. PMID- 15535256 TI - [Cytology of endometrial carcinoma--practice and actual problems]. PMID- 15535257 TI - [Histologic diagnosis of endometrial cancer]. PMID- 15535258 TI - [Endometrial cancer diagnosis by MR imaging]. PMID- 15535259 TI - [The value of ultrasonography in preoperative assessment of endometrial cancer]. PMID- 15535260 TI - [Outline of treatment for endometrial cancer]. PMID- 15535261 TI - [Surgical treatment of endometrial cancer]. PMID- 15535262 TI - [Surgical treatment for endometrial cancer]. PMID- 15535263 TI - [Individualization of surgery in endometrial cancer]. PMID- 15535264 TI - [Omission surgery in endometrial cancer]. PMID- 15535265 TI - [Recent outline of chemotherapy for uterine endometrial cancer]. PMID- 15535266 TI - [Update on postoperative chemotherapy for endometrial cancer]. PMID- 15535267 TI - [Adjuvant chemotherapy for high-risk endometrial carcinoma]. PMID- 15535268 TI - [Activity of paclitaxel, epirubicin and carboplatin in patients with advanced or recurrent endometrial cancer]. PMID- 15535269 TI - [Efficacy of cancer chemotherapy on patients with recurrent endometrial carcinoma]. PMID- 15535270 TI - [Hormone therapy for uterine corpus cancer--introduction]. PMID- 15535271 TI - [Conservative hormonal therapy for endometrial cancer]. PMID- 15535272 TI - [Postoperative hormone (endocrine) therapy in endometrial carcinoma]. PMID- 15535273 TI - [Radiotherapy for endometrial carcinoma]. PMID- 15535274 TI - [Sentinel lymph node detection in endometrial cancer]. PMID- 15535275 TI - [Para-aortic lymphadenectomy in endometrial carcinoma with cervical invasion]. PMID- 15535276 TI - [Prediction of nodal metastasis in endometrial carcinoma using positron emission tomography]. PMID- 15535278 TI - [Clinicopathological prognostic factors in endometrial carcinomas]. PMID- 15535277 TI - [PTEN expression in endometrial cancer and the prognosis]. PMID- 15535279 TI - [New pathologic staging which accurately predicts the prognosis of endometrial cancer]. PMID- 15535280 TI - [Long-time survival cases of endometrial cancer]. PMID- 15535281 TI - [Risk factors and prevention of uterine corpus cancer]. PMID- 15535282 TI - [Present status and problems in screening for endometrial cancer]. PMID- 15535283 TI - [Epidemiology of ovarian cancer]. PMID- 15535284 TI - [Molecular genetics of ovarian cancer]. PMID- 15535285 TI - [Genetic abnormalities in ovarian tumors]. PMID- 15535286 TI - [Overview of key genes implicated in ovarian carcinogenesis]. PMID- 15535287 TI - [Biological behavior of ovarian cancer]. PMID- 15535288 TI - [Mechanism of ovarian cancer metastasis--changes in E-cadherin expression]. PMID- 15535290 TI - [Pathology of ovarian cancer]. PMID- 15535289 TI - [Peritoneal dissemination of ovarian cancer and tumor angiogenesis]. PMID- 15535291 TI - [Borderline tumors of the ovary]. PMID- 15535292 TI - [General rules for management of ovarian cancers]. PMID- 15535293 TI - [Staging and management for ovarian cancer]. PMID- 15535294 TI - [Recent developments in the molecular genetic understanding of ovarian carcinoma]. PMID- 15535295 TI - [Tumor markers for ovarian cancers]. PMID- 15535296 TI - [Cytology of ovarian epithelial tumors]. PMID- 15535297 TI - [Pathological diagnosis of ovarian cancer]. PMID- 15535298 TI - [Diagnostic imaging for ovarian carcinoma]. PMID- 15535299 TI - [Therapy for ovarian cancer--general review]. PMID- 15535300 TI - [Surgical treatment for ovarian cancer]. PMID- 15535301 TI - [Wedge resection of the opposite ovary in conservative surgery of malignant ovarian tumor: is there any value?]. PMID- 15535302 TI - [Combined organ resection in cytoreductive surgery for ovarian cancer]. PMID- 15535303 TI - [Maximal cytoreductive surgery in patients with advanced ovarian cancer for improving the prognosis]. PMID- 15535304 TI - [Chemotherapy for ovarian cancer--introduction]. PMID- 15535305 TI - [Combination chemotherapy with docetaxel and carboplatin for epithelial ovarian cancer]. PMID- 15535306 TI - [Combination chemotherapy with nedaplatin and paclitaxel for ovarian cancer]. PMID- 15535307 TI - [Treatment of platinum-resistant ovarian cancer]. PMID- 15535308 TI - [Radiation therapy effects in ovarian cancer--review]. PMID- 15535309 TI - [Treatment of ovarian cancer with pelvic and paraaortic lymph node metastasis]. PMID- 15535310 TI - [Validity of complete paraaortic and pelvic lymphadenectomy in apparent stage I (T1) epithelial ovarian carcinoma]. PMID- 15535311 TI - [Gene expression and prognosis in ovarian cancer]. PMID- 15535312 TI - [Relationship between overexpression of HER-2/neu protein and prognosis in epithelial ovarian cancer]. PMID- 15535313 TI - [Prognostic importance of rupture of stage I ovarian cancer]. PMID- 15535314 TI - [Prognostic factors in advanced ovarian cancer patients]. PMID- 15535315 TI - [Prevention of ovarian cancer development]. PMID- 15535316 TI - [Mass-screening for ovarian cancer]. PMID- 15535317 TI - [Recent trends in multiple primary cancers associated with gynecological malignancy]. PMID- 15535318 TI - [Urinary, bowel, and sexual symptoms after surgery in gynecologic cancer]. PMID- 15535319 TI - [Ovarian cancers complicated by endometriosis]. PMID- 15535320 TI - [Kampo therapy for side effects of gynecologic carcinomatous chemotherapy]. PMID- 15535321 TI - [Palliative medicine for advanced gynecological malignancy]. PMID- 15535322 TI - [Clinical characteristics and therapeutic variations in elderly patients with ovarian cancer]. PMID- 15535323 TI - [Assessment of anxiety in cancer patients]. PMID- 15535324 TI - [Leg lymphedema after surgery for gynecologic cancer]. PMID- 15535325 TI - [Treatment of venous thromboembolism in cancer patients]. PMID- 15535326 TI - [Prophylaxis for post-operative thrombosis]. PMID- 15535327 TI - Tuberculosis in Malaysia: combating the old nemesis. PMID- 15535328 TI - Knowledge and practice of complementary medicine amongst public primary care clinic doctors in Kinta district, Perak. AB - The knowledge and practice of doctors (n=40) towards complementary medicine (CM) in 16 health clinics in the Kinta District were assessed by questionnaire. Thirty four (85%) responded. More than half felt that acupuncture (73.50), homeopathy (59%) and herbal medicine (59%) were occasionally harmful. Forty-four percent felt manipulative therapy was frequently harmful. Relaxation technique (79%) and nutritional therapy (44%) were considered most frequently useful. 59% used some form of CM. There were no significant differences found in usage rates by gender, age group and exposure to CM during undergraduate training. Sixty-seven percent had encouraged patients to seek CM. Seventy-three percent perceived an increasing demand for CM. Eighty-eight percent were in favour of a hospital based CM referral center. Only 6% were trained in CM. PMID- 15535329 TI - Coarctation of aorta repair at the National Heart Institute (1983-1994). AB - This retrospective study illustrates our approach to this problem over the years, from performing subclavian flap aortoplasty initially to the more accepted procedure now, which is resection and end-to-end anastomosis. Coarctation of aorta in our population is seen in a varying age groups and are also associated with other cardiac anomalies including both acyanotic and cyanotic congenital cardiac defects. Therefore a wide variety of surgical procedures were performed including resection of the coarcted segment and end-to-end anastomosis, subclavian flap aortoplasty, patch aortoplasty and synthetic tube graft interposition. Subclavian flap aortoplasty is not widely practised anymore in favour of resection with end-to-end anastomosis. Fifty four point four percent of patients had isolated coarctation, 10.5% had associated valvular defects, 28.1% had other simple congenital defects and 7.0% had associated complex cyanotic congenital defects. Perioperative mortality was 5.26% and is correlated with the younger age of patients at time of surgery and severity of cardiac failure at time of presentation. We did not see any difference in mortality for patients with complex congenital disease or between the different surgical procedures. However, we did find that in the early period when resection with end-to-end anastomosis was performed, there was a significantly higher incidence of morbidities. PMID- 15535330 TI - External fixation versus internal fixation for closed unstable intra-articular fracture of the distal radius. Early results from a prospective study. AB - This is a prospective study of a series of 26 patients with closed unstable comminuted intra-articular fracture distal end of the radius treated with two different methods of treatment to compare their anatomical and functional results. The external fixation group consisted of 12 patients and internal fixation group consisted of 14 patients. The anatomical and functional assessments were performed at six months and one year. The anatomical results at six months and one year showed that the internal fixation group was effective in maintaining the reduction compared to the external fixation group. The radial height, volar tilt and radial inclination were well maintained. However, the functional results at six months and one year showedno differences between these two types of fixation. The complication rate was higher in external fixation group. PMID- 15535331 TI - An evaluation of the tuberculosis control programme of Selangor State, Malaysia for the year 2001. AB - In the year 2001, 1459 Tuberculosis (TB) cases (43.1/100,000 population) were notified in Selangor. The highest age specific incidence rate was among those aged above 60 years and foreigners accounted for 15% of the cases notified. Fifteen percent of the TB cases were treated in the private sector where treatment efficacy and compliance could not be evaluated. Co- infection of Human Immunodeficiency Virus (HIV) infection with TB accounted for 51% of the TB deaths notified. Screening programmes in prisons and drug rehabilitation centres had detected 11.7% of HIV/TB coinfection among HIV positive inmates screened in these institutions. PMID- 15535332 TI - Inappropriate utilization of emergency department services in Universiti Sains Malaysia hospital. AB - Inappropriate utilization of Emergency Departments (ED) services may result in compromised management of patients requiring true emergency treatment. Significant attendance of non-emergency cases in ED was found in several countries. A cross-sectional study was conducted in Universiti Sains Malaysia Hospital (HUSM) to determine the proportion of the inappropriate cases and the utilization pattern by time (over 24 hours and within a week) and by diagnoses. A sample of 350 cases was randomly selected from ED-HUSM register of the year 2000. A decision flowchart, which was adopted from 4 guidelines, was applied to classify appropriate and inappropriate cases. There were 55% inappropriate cases in this study. The inappropriate cases increased considerably in early morning, late evening, during the weekend and early part of the week. Most common diagnoses of inappropriate cases were upper respiratory tract infections, mild acute gastroenteritis and urinary tract infections. Considerable attendance of inappropriate cases calls for interventions. PMID- 15535333 TI - Appropriateness of colonoscopy in a university hospital. AB - An open access endoscopy system allows for the direct scheduling of endoscopies by non-gastroenterologist physicians without prior gastroenterology consultation. The aim of our study was to examine our practice of open access endoscopy by evaluating the appropriateness of referrals for colonoscopy and to determine whether there were differences depending on the specialty of the referring clinician. The indication for colonoscopy was assessed in 499 consecutive outpatients referred for colonoscopy at University Hospital, Kuala Lumpur over a 12-month period. The American Society of Gastrointestinal Endoscopy (ASGE) guidelines were used to determine the appropriateness of referrals. 80.6% of colonoscopies requested by the gastroenterologist were performed for accepted indications compared to 50.6% of referrals by the primary care physician (p<0.001) and 67.0% of referrals by the surgeon (p=0.006). The rate of colonoscopies generally not indicated was 2.1% for the gastroenterologist, 25.0% for the internist (p=0.002) and 7.5% for the surgeon (p=0.04). The rate of indications not listed in the ASGE guidelines was significantly lower for requests made by gastroenterologists (17.3%) than those requested by primary care physicians (44.2%; p<0.001). Patients who have had prior consultation with the gastroenterologist were significantly more likely to undergo colonoscopy for appropriate indications than among patients who were referred through an open access system. The rate of inappropriate indications for colonoscopy was also significantly lower when the gastroenterologist made the referral. A substantial proportion of colonoscopies (25.4%) was performed for indications not listed in the ASGE guidelines. PMID- 15535334 TI - Does lifestyle increase the incidence of pregnancy-induced hypertension? AB - A case-control study was carried out in Alor Gajah to determine the socio economic, dietary and lifestyle factors and the occurrence of pregnancy-induced hypertension. There were a total of 30 cases who were selected from antenatal mothers attending 3 selected health centers in 1998. The control group consisted of 30 antenatal mothers who were matched according to health centre, race and age. The results showed that pregnancy-induced hypertension was significantly associated with obesity (P < 0.05) and being a housewife (P < 0.05). PMID- 15535335 TI - The prevalence of depression among the elderly in Sepang, Selangor. AB - OBJECTIVE: To determine the prevalence of depression among the elderly and to identify its associated factors. METHODS: A cross sectional study design was used. Stratified proportionate cluster sampling method was used to select the respondents. A 30 item Geriatric Depression Scale questionnaire was used as a screening instrument. RESULTS: 7.6% of the elderly were found to have depression. Only employment status was found to be significantly associated with depression. DISCUSSION: The prevalence of depression in the elderly was 7.6%. Primary Care providers need to be vigilant when treating elderly patients as depression is commonly found in this age group. PMID- 15535336 TI - An audit of intraoperative frozen section in Johor. AB - A 4-year-review was carried out on intraoperative frozen section consultations in Sultanah Aminah Hospital, Johor Bahru. Two hundred and fifteen specimens were received from 79 patients in the period between January 1999 and December 2002. An average of 2.72 specimens per patient was received. The overall diagnostic accuracy was high, 97.56%. The diagnoses were deferred in 4.65% of the specimens. False positive diagnoses were made in 3 specimens (1.46%) and false negative diagnoses in 2 specimens (0.98%). This gave an error rate of 2.44%. The main cause of error was incorrect interpretation of the pathologic findings. In the present study, frozen sections showed good sensitivity (97.98%) and specificity (97.16%). Despite its limitations, frozen section is still generally considered to be an accurate mode of intraoperative consultation to assist the surgeon in deciding the best therapeutic approach for his patient at the operating table. The use of frozen section with proper indications was cost-effective as it helped lower the number of reoperations. An audit of intraoperative frozen section from time to time serves as part of an ongoing quality assurance program and should be recommended where the service is available. PMID- 15535337 TI - Pulmonary tuberculosis--a review of clinical features and diagnosis in 232 cases. AB - The diagnosis of pulmonary tuberculosis is often delayed due to atypical clinical features and difficulty in obtaining positive bacteriology. We reviewed 232 cases of pulmonary tuberculosis diagnosed in Kedah Medical Centre, Alor Setar from January 1998 to December 2002. All age groups were affected with a male predominance (Male:Female ratio = 60:40). Risk factors include underlying diabetes mellitus (17.7%), positive family history (16.8%) and previous tuberculosis (5.2%). Nearly half (45.3%) of patients had symptoms for more than one year. Only 22% of patients had typical symptoms of tuberculosis (prolonged recurrent fever, cough, anorexia and weight loss), whilst others presented with haemoptysis, chronic cough, COPD, bronchiectasis, general ill-health, pyrexia of unknown origin or pleural effusion without other systemic symptoms. Fifteen percent of the patients presented with extrapulmonary diagnosis. Ninety percent of the patients had previous medical consultations but 40% had no chest radiograph or sputum examination done. The chest radiographs showed 'typical' changes of tuberculosis in 62% while in the other 38% the radiological features were 'not typical'. Sputum direct smear was positive for acid-fast bacilli in only 22.8% of patients and 11.2% were diagnosed base on positive sputum culture. Sputum may be negative even in patients with typical clinical presentations and chest radiograph changes. Bronchial washing improved the diagnosis rate being positive in 49.1% of cases (24.1% by direct smear and the other 25.0% by culture). In 16.8% of cases, the diagnosis was based on a good response to empirical anti-tuberculosis therapy in patients with clinical and radiological features characteristic of tuberculosis. In conclusions, the clinical and radiological manifestations of pulmonary tuberculosis may be atypical. Sputum is often negative and bronchoscopy with washings for Mycobacterium culture gives a higher yield for diagnosis. In highly probable cases, empirical therapy with antituberculosis drugs should be considered because it is safe and beneficial. PMID- 15535338 TI - Appropriate utilisation of emergency upper gastrointestinal endoscopy in a tertiary referral centre. AB - A retrospective cross-sectional study was carried out in a tertiary referral centre to determine the appropriateness of usage of emergency upper gastrointestinal endoscopy (EUGIE) with reference to the guidelines set by the American Society of Gastrointestinal Endoscopy (ASGE). EUGIE was defined as early, non-elective endoscopy performed for in-patients within 48 hours of acute hospital admission. The median age of the 668 patients was 55 years (age range 12 90), 31% of whom had a previous upper gastrointestinal endoscopy. Bleeding in the form of haematemesis, melaena or anaemia was the most common indication (40.7%) for EUGIE. Eighty one percent of the procedures were judged appropriate by the ASGE guidelines. There was a statistically significant relationship between appropriateness and significant diagnostic yield (P<0.05). Procedures performed for melaena, symptomatic anaemia and haemetemesis led to greater significant diagnostic yield (P<0.05) and there was no difference in the yield between working-hours and after-hours EUGIE. PMID- 15535339 TI - Risk factors for anti tuberculous drugs induced hepatitis: a prospective survey from a chest clinic in a general hospital. AB - A prospective survey on 14 consecutive cases with tuberculous drug induced hepatitis was done at our chest clinic in a state general hospital over a period of 15 months. There were 30 controls chosen randomly from the chest clinic register. The cases had lower mean body mass index (P<0.008), serum albumin (P<0.005) and higher serum globulin (P<0.04). Serum liver transaminases and total bilirubin rose significantly during the acute episode of drug induced hepatitis. Among the risk factors studied, only chronic hepatitis B carrier status was found to be more prevalent among the cases. There was one death (7.1%) over the whole study period. PMID- 15535340 TI - Assessment of knowledge, attitude and exposure to oncology and palliative care in undergraduate medical students. AB - We conducted a questionnaire survey among 261 year-4 and year-5 medical students containing 27 questions related to cancer, radiotherapy, general oncology and palliative care to assess their knowledge, understanding, and exposure to oncology and palliative care in our medical school. Out of 261 students, 139 students returned their questionnaire for analysis. Twenty nine percent (29%) of the students had rarely visited the Radiotherapy and Oncology unit. There were profound deficiencies in the basic knowledge of cancer (46%), principles of radiotherapy treatment (59%), palliative care (64%), and cancer prevention (48%). They reported no specific teaching about early detection of common malignancies and cancer prevention. The main input of instruction about cancer came from surgery (46%) and pathology (28%) teachings. This study revealed that there is deficiency in cancer education in the undergraduate teaching program in our institution. PMID- 15535341 TI - Developing the Malaysian health system to meet the challenges of the future. AB - The Malaysian health care system is a success story among countries of equivalent socio-economic status. However there are numerous challenges faced by the nation, which create the need for changes and reform. There is rising consumer demands and expectations for high technology and high cost medical care due to improved standards of living, changing disease patterns and demographic changes, inadequate integration of health services, maldistribution of resources and the threats as well as opportunities of globalisation and liberalisation. The changes in health policy, priorities and planning for the country are guided by the country's development policies, objectives and the challenges of Vision 2020, Vision for Health and the goals of the health system in ensuring universal access, improving equity and efficiency and the quality of life of the population. The essential services in the health system of the future are information and education of individuals to empower support for the wellness paradigm. There is also a need to restructure the national health care financing and the health care delivery system. The present roles and responsibilities of MOH also need to be reviewed. PMID- 15535342 TI - Folic acid in the prevention of neural tube defect--a programme for Malaysia? AB - There is good evidence that folic acid is safe and efficacious for reducing neural tube defect (NTD). All women capable of becoming pregnant should take 400 microgram daily. This can be given in the form of a daily vitamin supplement, by food fortification, or by increasing natural dietary folates. Compulsory grain fortification has been shown to reduce NTD in a population but supplementation and dietary advice have not. Malaysia should work towards a programme of grain fortification and use alternative strategies to reach sections of the population that would not be covered but several research questions need to be answered before a programme could be implemented in Malaysia. PMID- 15535343 TI - Familial hemophagocytic lymphohistiocytosis in two brothers. AB - Familial hemophagocytic lymphohistiocytosis is a disorder which presents with fever, pancytopenia, liver dysfunction and also an increase in non-malignant histiocytes with prominent hemophagocytosis in various organs. It is usually difficult to distinguish from other hemophagocytic syndrome in the absence of family history. It rarely manifests in adults. Chemotherapy is usually indicated. Here, we report the occurrence of this disorder in two brothers in their twenties. PMID- 15535344 TI - Intravenous immunoglobulin in the treatment of acute disseminated encephalomyelitis. AB - We describe a case of acute disseminated encephalomyelitis in a child. This case is unusual in that the illness was characterised by recurrent episodes rather than a monophasic course and that the choice of treatment was intravenous immunoglobulin over corticosteroids. The rapid and remarkable recovery is highlighted and a review of the treatment for this rare condition is discussed. PMID- 15535345 TI - Concurrent mycobacterial infection and non-Hodgkin's lymphoma at the same site in an AIDS patient. AB - A 31 years old Chinese male with acquired immunodeficiency syndrome (AIDS) presented with concurrent mycobacterial infection and a synchronous non-Hodgkin's lymphoma of the nose. The diagnoses were made over a period of two months. Treatment for the mycobacterial infection was administered but he succumbed to the disease shortly after the diagnosis of NHL was established. This was an unusual case where two pathologies occurred in the same site in a patient with AIDS. PMID- 15535346 TI - Pulmonary resection for metastatic breast cancer. AB - The management of pulmonary metastasis from breast carcinoma is challenging and often consists of palliation of symptoms. Surgical resection of pulmonary metastasis is considered inappropriate in view of the disseminated nature of the disease and limited life expectancy. It can however be a worthwhile option if imaging, including bone scans rule out metastatic disease in other part of the body. We report a patient with pulmonary metastasis from breast carcinoma who was successfully treated with pulmonary wedge resection of the metastatic lesion. PMID- 15535347 TI - Rare cause of Horner's syndrome: pseudoaneurysm of right subclavian artery in an intravenous drug user. AB - Pseudoaneurysms of the subclavian artery are extremely rare lesions despite the overall increase in the frequency of septic pseudoaneurysms caused by illicit parenteral drug abuse. A case of subclavian artery pseudoaneurysm presenting with Horner's syndrome in an intravenous drug user is discussed. PMID- 15535348 TI - Tuberculous orchitis in chronic renal failure. AB - Tuberculosis confined to the testes with no epididymal involvement is uncommon. Chronic renal failure patients requiring hemodialysis have increased risk for developing tuberculosis. We report a 47-year old chronic renal failure man presenting with right testicular tuberculous orchitis. A high index of suspicion is required to recognize the unusual presentation of tuberculosis in this group of patients, and routine screening for tuberculosis may be recommended in patients undergoing hemodialysis. PMID- 15535349 TI - Spontaneous closure of a persistent post-traumatic pancreatic fistula in a child: incidental therapeutic effect of ERCP. AB - A child with a post-tramatic pancreatic pseuodocyst developed pancreatic fistula, following percutaneous drainage of the cyst. The fistula output was about 120ml/day and lasted for a month. A diagnostic ERCP revealed main pancreatic duct disruption. The fistula output ceased over the next two days following the ERCP. The patient has been well for the past four years. This patient's immediate recovery after ERCP could possibly be due to dislodgement of a mucus plug by the contrast injected during ERCP. Such a mechanism can explain the previous sporadic reports of spontaneous cure of persistent pancreatic fistulae in children. PMID- 15535350 TI - Giant aneurysm or pituitary macroadenoma: a diagnostical misconstrue. AB - We describe a case of a giant thrombosed intrasellar internal carotid artery aneurysm initially diagnosed as pituitary macroadenoma. The differentiating neuroimaging features were discussed. PMID- 15535351 TI - Neonatal post-intubation subglottic stenosis. AB - Acquired subglottic stenosis is a compication or neonatal endotracheal intubation. Although it is rare, it contributes significantly to the morbidity and physical well being of post extubated neonates. A 20-day old neonate, ventilated for meconium aspiration syndrome and persistent fetal circulation, presented with marked stridor and respiratory embarrassment. A stenosed subglottic area was found on rigid bronchoscopy. Anterior cricoid split was performed to relieve the obstruction. He is asymptomatic post operatively. PMID- 15535352 TI - Are doctors in the university-based primary care clinic more evidence-based? PMID- 15535353 TI - What is obesity? PMID- 15535354 TI - A needed specialty. PMID- 15535355 TI - The smallest sufferers: caregivers and parents must truly understand children's pain to treat it effectively. PMID- 15535356 TI - Back in the game: a four-phase return-to-sport program for athletes with problem ACLS. PMID- 15535357 TI - Only the best: today's best seating and mobility practices may be tomorrow's standards. AB - In the area of seating and mobility evaluations, best practices can best be depicted and illustrated by a wheel similar to one found on a wheelchair. In the center or axle of the wheel is the concept of best practices. Spreading out from this center are the spokes that stabilize and support the wheel. The wheel requires that all of the spokes be in good working order for it to function properly. These spokes of best practices include experience, hands-on techniques, skills, technology, resources, self-directed learning, and the consumer. All the spokes must be strong and in place in order for the wheel to run well. Not only does a wheel have spokes, it also has a brake. This brake has the potential to slow or even stop the wheel. However, if the brake is not applied, it has no effect on the wheel. When the wheel is functioning properly, it can be put into motion. PMID- 15535358 TI - Hands-on experience: specific hand conditions affect the aging population. PMID- 15535359 TI - Ultrasound: underused? PMID- 15535360 TI - The value of percentages. PMID- 15535362 TI - Starry night: a texture devoid of depth cues. AB - From a modern Bayesian point of view, the classic Julesz random-dot stereogram is a cue-conflict stimulus: Texture cues specify an unbroken, unslanted surface, in conflict with any variation in depth specified by binocular disparity. We introduce a new visual texture-the starry night texture (SNT)--that is incapable of conveying slant, depth edges, or texture boundaries, in a single view. For SNT, changing density is equivalent to changing intensity, so an instance of the texture is characterized (up to the random locations of the texture elements) by what we call its densintensity. We consider deviations from the ideal that are needed to realize the texture in practice. In three experiments with computer generated stimuli we examined human perception of SNT to show that (1) the deviations from the ideal that were needed to realize SNT do not affect the invariance of its appearance across changes in distance of several orders of magnitude; (2) as predicted, observers match SNT across changes in distance better than other textures; and (3) the use of SNT in a slant perception experiment did not reliably increase observers' reliance on stereoscopic slant cues, as compared with the sparse random-dot displays that have been commonly employed to study human perception of shape from binocular disparity and motion. PMID- 15535361 TI - Back to back: a comparison of the health care proposals of the presidential candidates. PMID- 15535363 TI - On the optical theory of underwater vision in humans. AB - Defocus changes the visual contrast sensitivity function, thereby creating a complex curve with local dips and peaks. Since underwater vision in humans is severely defocused, we used optical theory and the phenomenon of spurious resolution to predict how well humans can see in this environment. The values obtained correspond well with experimental measurements of underwater human acuity from earlier studies and even point to an opportunity for humans with exceptional contrast sensitivity to see better underwater than the children in those studies. The same theory could be useful when discussing the visual acuity of amphibious animals, as they may use pupil constriction as a means of improving underwater vision. PMID- 15535364 TI - Gaussian weighting of ocular wave-front measurements. AB - The measurement of ocular wave-front error gives insight into the optical performance of the eye and possibly a means for assessing visual performance. The visual system responds not only to the quality of the optical image formed on the retina but also to the processing that occurs in the retina and the brain. To develop a metric of visual performance based on wave-front error measurements, these latter processes must somehow be incorporated. In representing the wave front error in terms of Zernike polynomials, it appears that terms with lower angular frequency have a greater deleterious effect on visual performance than higher-angular-frequency terms. A technique for weighting the pupil function of the eye with a Gaussian filter is demonstrated. It is further demonstrated that the variance of the Gaussian-weighted wave-front error is well correlated with visual performance. PMID- 15535365 TI - Fisher-information-based analysis of a phase-diversity-speckle imaging system. AB - The question of just how much to defocus the second image in the conventional two channel phase-diversity speckle imaging technique may be addressed in a number of ways. Fisher information furnishes a useful metric for optimizing the choice of defocus as a functional of the object class, operating conditions, and the imaging task. Approximate closed-form expressions for the Fisher information relative to object parameters, rather than the pupil phase, are derived and discussed for phase-diversity-speckle imaging under conditions of strong turbulence and additive Gaussian noise. As an application of our general information-theoretic approach, the optimization of defocus when the imaging task is to estimate the midfrequency power spectrum of the object is discussed. PMID- 15535366 TI - Intrinsic orbital angular momentum of paraxial beams with off-axis imprinted vortices. AB - We investigate the orbital angular momentum (OAM) of paraxial beams containing off-axis phase dislocations and put forward a simple method to calculate the intrinsic orbital angular momentum of an arbitrary paraxial beam. Using this approach we find that the intrinsic OAM of a fundamental Gaussian beam with a vortex imprinted off axis has a Gaussian dependence on the vortex displacement, implying that the expectation value of the intrinsic OAM of a photon can take on a continuous range of values (i.e., integer and noninteger values in units of h). Finally, we investigate, both numerically and experimentally, the far-field profiles of beams carrying half-integer OAM per photon, these beams having been created by the method of imprinting off-axis vortices. PMID- 15535367 TI - Spatial correlation properties of focused partially coherent light. AB - We investigate the spatial coherence properties in the focal region of a converging, spatially partially coherent wave field. In particular, we find that, depending on the effective coherence length of the field in the aperture, the longitudinal and transverse coherence lengths in the focal region can be either larger or smaller than the corresponding width of the intensity distribution. Also, the correlation function is shown to exhibit phase singularities. PMID- 15535368 TI - Phase singularities of the longitudinal field components in the focal region of a high-aperture optical system. AB - We study the behavior of the longitudinal components of strongly focused electromagnetic fields. These components possess phase singularities and phase saddles that can be annihilated when the aperture angle of the lens is changed. PMID- 15535369 TI - Physical properties of wave scattering by a chiral grating. AB - Wave scattering by a chiral grating is studied in this paper. Numerical results are given, and physical properties are discussed, including the influence of frequency, angle of incidence, and aspect ratio. At high frequencies we find anomalous coupling regions known as Wood's anomalies, which are explained by the excitation and reradiation of leaky waveguide modes in the periodic layer. The chiral grating can possess both frequency-selection and mode-conversion properties. PMID- 15535371 TI - Entropy of partially polarized light and application to statistical processing techniques. AB - We have analyzed entropy properties of coherent and partially polarized light in an arbitrary number of spatial dimensions. We show that for Gaussian fields, the Shannon entropy is a simple function of the intensity and of the Barakat degree of polarization. In particular, we provide a probabilistic interpretation of this definition of the degree of polarization. Using information theory results, we also deduce some physical properties of partially polarized light such as additivity of the entropy and depolarization effects induced by mixing partially polarized states of light. Finally, we demonstrate that entropy measures can play an important role in segmentation and detection tasks. PMID- 15535370 TI - Overall coherence and coherent-mode expansion of spectrally partially coherent plane-wave pulses. AB - The modal theory for spectrally partially coherent nonstationary plane waves is introduced. The theory is first developed in the space-frequency domain and then extended to the space-time domain. Propagation properties of the coherent modes are analyzed. The concept of the overall degree of coherence is extended to the domain of nonstationary fields, and it is shown that the overall degree of coherence of partially coherent plane-wave pulses is the same in the space frequency and space-time domains. The theory is applied to the recently introduced concept of spectrally Gaussian Schell-model plane-wave pulses. PMID- 15535372 TI - Fast method for physical optics propagation of high-numerical-aperture beams. AB - A method is presented that expands the scheme of physical optics propagation beyond the Fresnel approximation to include beams that are nonparaxial. The formalism retains most of the calculation advantages of the Fresnel approach; i.e., it is based on a single Fourier transform step. The kernel of the new transformation is no longer separable in Cartesian coordinates; thus the formalism can account for astigmatic coupling effects originating purely from diffraction. The validity limits of the proposed algorithm are explored. Analytical expressions, numerical simulation results, and experimental data are compared. PMID- 15535373 TI - Generalized optical ABCD theorem and its application to the diffraction integral calculation. AB - We generalize the transfer matrix ABCD theorem for paraxial rays of the optical system to skew rays propagated off axis, whether or not the system possesses rotational symmetry. Furthermore, we apply the generalized ABCD theorem to evaluate the diffraction integral matrix elements A-D expressed in terms of the angle eikonal T, with the primary aberrations included. Finally, analysis and numerical calculation are given for propagation of a light beam through the optical system in the case in which spherical aberration and coma are present. PMID- 15535374 TI - A variational principle in optics. AB - We derive a new variational principle in optics. We first formulate the principle for paraxial waves and then generalize it to arbitrary waves. The new principle, unlike the Fermat principle, concerns both the phase and the intensity of the wave. In particular, the principle provides a method for finding the ray mapping between two surfaces in space from information on the wave's intensity there. We show how to apply the new principle to the problem of phase reconstruction from intensity measurements. PMID- 15535375 TI - Phase-space interpretation of deterministic phase retrieval. AB - Deterministic phase retrieval is reinterpreted in terms of phase-space optics. A novel derivation of the transport-of-intensity equation is presented based on the Wigner distribution function and the ambiguity function. The phase retrieval problem is formulated as estimating the local first-order moment of the Wigner function from intensity information. A comparison with phase-space tomography suggests a generalization of deterministic phase retrieval that provides larger flexibility for signal recovery. In addition, one particular numerical implementation of generalized deterministic phase retrieval is presented. Simulated intensity data are used to validate the method. PMID- 15535376 TI - Ultracompact splitter for submicrometer silicon-on-insulator rib waveguides. AB - An ultracompact and efficient 1 x 2 splitter for submicrometer silicon-on insulator rib waveguides using a star coupler is reported. The structure proposed here is decidedly smaller than the usual splitters such as multi-mode interference or Y-branch devices and much less sensitive to technological fluctuations. Design of the compact splitter is optimized at lambda = 1.31 microm with the effective-index method and a two-dimensional beam-propagation method. The excess losses are lower than 0.15 dB, and the dependence of the losses on wavelength between 1.23 and 1.63 microm is almost flat (variation less than 0.04 dB), which makes the device very interesting for coarse wavelength-division multiplexing applications within the silicon photonic technology. PMID- 15535377 TI - Comparative study of three methods for the simulation of two-dimensional photonic crystals. AB - Three methods for the efficient simulation of two-dimensional photonic crystal structures are compared, namely, a semianalytical multiple-scattering technique; a vectorial eigenmode expansion technique; and a FDTD-ROM technique. The basic principles of each method are presented. For the semianalytical technique and for the vectorial eigenmode expansion technique, we show how reflections coming from abruptly terminated waveguides can be avoided. The main advantages and disadvantages of each method are discussed. Results from use of the three methods are compared for several photonic crystal structures. PMID- 15535378 TI - Explicit analysis of anisotropic planar waveguides by the analytical transfer matrix method. AB - The propagation properties of light in anisotropic optical planar waveguides with different index distributions are investigated with the analytical transfer matrix method. Dispersion equations are analytically deduced by the method in terms of different index profiles. It is shown by examples that this method exhibits good accuracy compared with numerical methods while still holding physical insight. PMID- 15535379 TI - Theory of partially coherent electromagnetic fields in the space-frequency domain. AB - We construct the coherent-mode representation for fluctuating, statistically stationary electromagnetic fields. The modes are shown to be spatially fully coherent in the sense of a recently introduced spectral degree of electromagnetic coherence. We also prove that the electric cross-spectral density tensor can be rigorously expressed as a correlation tensor averaged over an appropriate ensemble of strictly monochromatic vectorial wave functions. The formalism is demonstrated for partially polarized, partially coherent Gaussian Schell-model beams, but the theory applies to arbitrary random electromagnetic fields and can find applications in radiation and propagation and in inverse problems. PMID- 15535380 TI - Nonradiating surface sources. AB - The class of three-dimensional (3D) nonradiating sources to the inhomogeneous Helmholtz equation is generalized to include nonradiating surface sources that are localized on closed or infinite surfaces sigma and that generate no radiation throughout one of the two space regions whose common boundary is sigma. The class of non-radiating surface sources is shown to be identical to the class of so called secondary surface sources that are generated from boundary values of a field radiated by a 3D source. In particular, it is proven that any secondary surface source radiating into the interior (exterior) of sigma is a nonradiating surface source relative to the exterior (interior) of sigma, and vice versa. Examples of nonradiating surface sources are included, and a discussion is given of possible applications of such sources. PMID- 15535381 TI - Optimization of photonic crystal structures. AB - We report on the numerical structural optimization of two-dimensional photonic crystal (PhC) power dividers by using two different classes of optimization algorithms, namely, a modified truncated Newton (TN) gradient search as deterministic local optimization scheme and an evolutionary optimization representing the probabilistic global search strategies. Because of the severe accuracy requirements during optimization, the proper PhC device has been simulated by using the multiple-multipole program that is contained in the MaX-1 software package. With both optimizer classes, we found reliable and promising solutions that provide vanishing power reflection and perfect power balance at any specified frequency within the photonic bandgap. This outcome is astonishing in light of the discrete nature inherent in the underlying PhC structure, especially when the optimizer is allowed to intervene only within a very small volume of the device. Even under such limiting constraints structural optimization is not only feasible but has proven to be highly successful. PMID- 15535382 TI - Wigner functions for nonparaxial, arbitrarily polarized electromagnetic wave fields in free space. AB - New representations are defined for describing electromagnetic wave fields in free space exactly in terms of rays for any wavelength, level of coherence or polarization, and numerical aperture, as long as there are no evanescent components. These representations correspond to tensors assigned to each ray such that the electric and magnetic energy densities, the Poynting vector, and the polarization properties of the field correspond to simple integrals involving these tensors for the rays that go through the specified point. For partially coherent fields, the ray-based approach provided by the new representations can reduce dramatically the computation times for the physical properties mentioned earlier. PMID- 15535383 TI - Interferometric imaging polarimeter. AB - We describe a new imaging polarimeter that is based on a modified Sagnac interferometer and allows full polarimetric description of complex random electromagnetic beams from only two images as compared with the four images needed in conventional polarimetry. The procedure is analyzed in terms of the interference laws of Fresnel and Arago, and very good agreement with standard Stokes polarimetry is demonstrated. In certain cases the new technique provides the degree of polarization and the retardance from only one image. PMID- 15535384 TI - National study of adolescent mental health points to missing role of nurses. PMID- 15535385 TI - Childhood externalizing behavior: theory and implications. AB - TOPIC: Childhood externalizing behavior PURPOSE: To analyze the construct of externalizing behavior (aggression, delinquency, and hyperactivity), illustrate the biosocial model of childhood externalizing, and draw clinical implications for nursing research and practice. SOURCES: A review of the literature based on psychological, psychiatric, and nursing journals. CONCLUSIONS: A better understanding of childhood externalizing behavior problems and the risk factors underlying them are essential to prevent them. The employment of an integrative biosocial perspective is argued to be important in understanding this behavior. PMID- 15535386 TI - Early reactions to frustration: developmental trends in anger, individual response styles, and caregiving risk implications in infancy. AB - PROBLEM: In an effort to examine early negative emotional response patterns, infant responses to frustration were studied with specific regard to anger. METHODS: Forty infants, 7-9 months (n = 20) and 12-14 months (n = 20) old, were observed in laboratory play situations. Infant affective response as well as maternal behavior were observed. Maternal interviews assessed maternal impressions of infant behavior and affect. FINDINGS: Findings revealed increased displays of anger in older infants, evidence of cognitive-affective linkages in response, and individual differences in emotional response style and maternal infant processes. CONCLUSIONS: Findings relate to the emergence and interplay of negative emotional response, maternal-infant patterns, and socioemotional risk. PMID- 15535387 TI - Level systems: inpatient programming whose time has passed. AB - TOPIC: Structuring of inpatient behavioral programming in child-adolescent psychiatric, residential treatment, and juvenile justice settings. PURPOSE: To review the underlying theory underpinning current practices and recommend remedies to the uncovered problems. SOURCES: A review of the literature from 1965 to 2001 from selected nursing and medical psychiatric and mental health publications. CONCLUSIONS: Intensive professional and staff education and greater precision in communication about patients' behaviors are needed in many settings. There is also a need to move away from generic treatment approaches and return to individual treatment planning based on individual assessments and the unique needs of an increasingly volatile and complex in-patient population. PMID- 15535388 TI - Coping with the pediatric anogenital exam. AB - PURPOSE: To focus on children's and adult guardians' anticipatory and acute attempts to cope with the pediatric anogenital exam. METHODS: A sample of 40 girls (ages 8-15) and a separate sample of 109 adult guardians who accompanied children to the medical clinic rated their use of specific coping strategies on a 5-point Likert scale both pre- and postexam. FINDINGS: Overall, participants reported relatively high levels of active coping strategies and relatively low levels of passive coping responses throughout the medical encounter. Effects of situational and dispositional variables on the use of specific coping strategies also were found. CONCLUSIONS: Data provided indirect evidence of the inherent strengths and self-righting tendencies of those dealing with the exceptional circumstances surrounding allegations of child sexual abuse. Such data suggest the value of identifying individuals' coping resources and options to facilitate successful completion of the medical evaluation and provide therapeutic benefit to patients and families. PMID- 15535389 TI - A simple numerical procedure for estimating nonlinear uncertainty propagation. AB - Accurately determining the effect of the propagation of uncertainty in nonlinear applications can be awkward and difficult. The Monte Carlo approach requires statistically significant numbers of function evaluations (typically 10(5) or more) and analytical methods are intractable for all but the simplest cases. This paper derives and demonstrates a method to estimate the propagation of uncertainty in nonlinear cases by representing the function in a piecewise fashion with straight line segments. The probability density function of the result can be calculated from the transformation of the line segments. The mean and confidence intervals of the result can then be calculated from the probability density function. For the special case of a normal distribution in the independent variable, calculation of the mean and confidence intervals requires evaluation of only the error function (erf). A simple example is presented to demonstrate the technique. Variations on the basic approach are presented and discussed. PMID- 15535390 TI - Solution of low-dimensional constrained model predictive control problems. AB - Large benefits are possible by utilizing the solution of the constrained optimization problem involved in model predictive control. For a special case of these problems, the solution can be obtained relatively easily from its relationship with the unconstrained optimum. In this paper, a visualization of the relationship between the constrained and unconstrained optimum is presented. Based upon this relationship, a method for finding the constrained optimum is proposed that is suitable for low-dimensional control systems. A comparison with a linear programming formulation on 2 x 2 and 3 x 3 problems shows that the computational effort can be 10-35 times lower. For such processes, the proposed approach may allow one to avail the benefits of optimization by using the small process control systems already present in many plants. PMID- 15535391 TI - Comparison of some modeling and control issues for a flexible two link manipulator. AB - The nonlinear dynamics of a flexible two link manipulator presents a challenging modeling and control problem with its lumped actuator dynamics coupled with distributed link dynamics. Issues related to modeling uncertainty and control robustness as well as a good understanding of the experimental issues are essential for advanced modeling and control for such systems. This paper reports some of the advanced modeling, control, and experimental issues for a flexible two link manipulator, including some novel approaches developed by the authors. PMID- 15535392 TI - Position tracking control of saturated LSRM. AB - This paper deals with the tracking control design of a linear synchronous reluctance motor (LSRM) drive. An extended nonlinear dynamic LSRM model with magnetic saturation included is used in the control design and practical realization, in order to improve tracking performances at very low speeds of motion. Iron core saturation is included in the extended model with the experimentally determined flux linkages given as functions of the direct and quadrature axes currents. Experimental results show that the proposed input output linearizing tracking control with included saturation behaves considerably better than the one without saturation, introducing smaller position and speed errors and better motor stiffness, on account of the increased computational complexity. PMID- 15535393 TI - Low order robust controller design for preserving Hinfinity performance: genetic algorithm approach. AB - This paper investigates the design of low order robust controllers based on an Hinfinity performance index using a real-code genetic algorithm. In Hinfinity controller design, the major disadvantage of the existing methods is that they lead to high-order controllers. This is the gap between theory and practice. Therefore the purpose of this paper is to design a low order controller with similar performance to the Hinfinity optimal controllers, which can find sufficiently wide use in engineering practice. We first design the Hinfinity optimal controller using Glover and Doyle's results, and obtain the corresponding performance index gamma. Second, the desired low order controller with several parameters is chosen, e.g., a first-order controller, or a PID controller. Finally, we use the real-code genetic algorithm to find the optimal controller parameters that preserve the performance index y. Computational simulations illustrate the effectiveness of the proposed approach. PMID- 15535394 TI - Intelligent on-line fault tolerant control for unanticipated catastrophic failures. AB - As dynamic systems become increasingly complex, experience rapidly changing environments, and encounter a greater variety of unexpected component failures, solving the control problems of such systems is a grand challenge for control engineers. Traditional control design techniques are not adequate to cope with these systems, which may suffer from unanticipated dynamic failures. In this research work, we investigate the on-line fault tolerant control problem and propose an intelligent on-line control strategy to handle the desired trajectories tracking problem for systems suffering from various unanticipated catastrophic faults. Through theoretical analysis, the sufficient condition of system stability has been derived and two different on-line control laws have been developed. The approach of the proposed intelligent control strategy is to continuously monitor the system performance and identify what the system's current state is by using a fault detection method based upon our best knowledge of the nominal system and nominal controller. Once a fault is detected, the proposed intelligent controller will adjust its control signal to compensate for the unknown system failure dynamics by using an artificial neural network as an on-line estimator to approximate the unexpected and unknown failure dynamics. The first control law is derived directly from the Lyapunov stability theory, while the second control law is derived based upon the discrete-time sliding mode control technique. Both control laws have been implemented in a variety of failure scenarios to validate the proposed intelligent control scheme. The simulation results, including a three-tank benchmark problem, comply with theoretical analysis and demonstrate a significant improvement in trajectory following performance based upon the proposed intelligent control strategy. PMID- 15535395 TI - Tuning of PID controllers for boiler-turbine units. AB - A simple two-by-two model for a boiler-turbine unit is demonstrated in this paper. The model can capture the essential dynamics of a unit. The design of a coordinated controller is discussed based on this model. A PID control structure is derived, and a tuning procedure is proposed. The examples show that the method is easy to apply and can achieve acceptable performance. PMID- 15535396 TI - An approach to predictive control of multivariable time-delayed plant: stability and design issues. AB - In this paper, a new method of multivariable predictive control is presented. The main advantage of a predictive approach is that multivariable plants with time delays can be easily handled. The proposed control algorithm also introduces a compact and simple design in the case of higher-order and nonminimal phase plants, but it is limited to open-loop stable plants. The algorithm of the proposed multivariable predictive control is developed, designed, and implemented on an air-conditioned system. The stability of the proposed control law is discussed. PMID- 15535397 TI - Analysis and performance of the control systems of the NASA 70-meter antennas. AB - The control systems of the NASA 70-m antennas include the antenna control system, the Master Equatorial (ME) control system, and their combinations (called modes). The Master Equatorial is a small telescope mounted on the top of a tower located inside the antenna structure. In the Antenna Encoder mode antenna encoders are used to close the feedback loop. In the Autocollimator mode the Master Equatorial is a master that follows a target, and the antenna is a slave that follows the Master Equatorial. In the Master Equatorial Encoder mode the "master-slave" relationship is reversed. In the paper the analysis begins with the description of the open-loop models of the antenna and of the Master Equatorial. We obtained the models by using field test data and system identification techniques. Next, we analyzed and evaluated the performance of the three modes of the antenna control system. The analysis showed that the Autocollimator and Master Equatorial Encoder tracking modes are feasible for high-rate tracking, and that the latter mode has the smallest tracking error. Finally, we analyzed the switching between antenna modes, necessary while tracking near the keyhold. We showed that switching causes jerks of magnitudes within the acceptable threshold. The contribution of this paper includes the development of the antenna model using field data and system identification procedures, the development of the LQG control algorithm for the 70-meter antenna, the development of two control cooperating systems (antenna and ME), identifying the more appropriate, and analyzing of switching between the two control systems. PMID- 15535398 TI - Comparison of two approaches to automated PI controller tuning for an industrial weigh belt feeder. AB - In this paper, two advanced PI controller tuning methods, unfalsified control and fuzzy control, are applied to an industrial weigh belt feeder that has significant nonlinearities. Both methods do not require an explicit plant model. The advantage of the unfalsified PI control design method is that it is able to directly incorporate multiple performance criteria, while the advantage of fuzzy logic is that it is able to directly incorporate human reasoning in the design process. Experimental results exhibit the effectiveness of both control methods. A detailed comparison of the two approaches is given in the areas of allowed design specifications, process knowledge requirements, computational requirements, controller development effort, transient performance, and the ability to handle motor saturation. PMID- 15535399 TI - Worst-case response time of aperiodc message in WorldFIP and its improvement in real-time capability. AB - How to support real-time traffic of aperiodic messages using WorldFIP is investigated in this paper. Transmission of aperiodic message within WorldFIP is a very complicated procedure because it is concerned with left time in a microcycle and must successively experience three different stages, indicating, identifying, and transmitting aperiodic message. This paper presents an integrated approach to analyze the worst-case responding time (WCRT) of aperiodic message, through integrating transmission of periodic message and these three stages of aperiodic message together. To decrease pessimistic factors in WCRT of aperiodic message, priority of aperiodic message is considered. Detail analysis of the WCRT of aperiodic message indicates that the current scheduling method cannot effectively utilize priority to guarantee important aperiodic messages. Then improved scheduling methods, one adding priority and another adjusting the schedule table, are proposed. The improved methods are finally validated by case studies. PMID- 15535400 TI - Graphical enhancement to support PCA-based process monitoring and fault diagnosis. AB - Principal component analysis (PCA) for process modeling and multivariate statistical techniques for monitoring, fault detection, and diagnosis are becoming more common in published research, but are still underutilized in practice. This paper summarizes an in-depth case study on a chemical process with 20 monitored process variables, one of which reflects product quality. The analysis is performed using the PLS_Toolbox 2.01 with MATLAB, augmented with software which automates the analysis and implements a statistical enhancement that uses confidence limits on the residuals of each variable for fault detection rather than just confidence limits on an overall residual. The newly developed graphical interface identifies and displays each variable's contribution to the faulty behavior of the process; and it aids greatly in analyzing results. The case study analyzed within shows that using the statistical enhancement can reduce the fault detection time, and the automated graphical interface implements the enhancement easily. PMID- 15535401 TI - Clinical outcome to interferon therapy in chronic hepatitis C: insights from mechanistic studies. PMID- 15535402 TI - HIV drug resistance testing: is the evidence really there? AB - OBJECTIVES: To assess the strength of evidence supporting the routine use of HIV drug resistance testing. DESIGN: A critical review of all studies relating to the clinical utility of HIV resistance testing, with a focus on randomized trials. RESULTS: Two cohort studies found no evidence of a difference in virological response in patients who had resistance testing compared with matched controls. We identified nine published randomized trials that were specifically designed to assess the clinical utility of drug resistance testing. In a meta-analysis of these trials, resistance testing increased the proportion of patients who achieved undetectable viral load by an average of 7% (95% confidence interval: 3 11%). However, this may be an over-estimate of the impact of resistance testing in clinical practice because of the idealized design and analytical approaches used in most of the studies. CONCLUSIONS: The available evidence does not clearly demonstrate that HIV drug resistance testing is clinically effective. To optimize their value for health decision-making, future trials of HIV resistance testing should be carefully designed to mimic the circumstances of routine clinical practice. PMID- 15535403 TI - Management of dyslipidaemia in HIV-infected patients receiving antiretroviral therapy. AB - Dyslipidaemia associated with the treatment of HIV infection, particularly with the use of protease inhibitors (PIs), can raise cholesterol and triglyceride (TG) levels to the thresholds indicated for intervention. Recent evidence from epidemiological studies has shown that there are correlations between antiretroviral drug use and increased risks for, and incidences of, cardiovascular disease, including myocardial infarction and coronary heart disease. The primary goals of dyslipidaemia therapy for HIV patients are reductions of both low-density lipoprotein cholesterol (LDL-C) and markedly elevated TG levels. Dietary strategies and exercise programs may be tried, although these have shown inconsistent results. The two options for drug therapy are switching antiretroviral agents and using lipid-lowering drugs. Each approach is associated with advantages and limitations, and the need to maintain viral suppression must be balanced with the need to treat abnormal lipid levels. Most drug switches replace the PI component with drugs from another antiretroviral class. Selection of drug therapy for lipid lowering depends on the type of dyslipidaemia predominating and the potential for drug interactions. The use of the statins pravastatin and atorvastatin is recommended for the treatment of patients with elevated LDL-C levels and gemfibrozil or fenofibrate for patients with elevated TG concentrations. Development of new PIs with more favourable effects on the lipid profile should be of benefit. PMID- 15535404 TI - HPV proteins as targets for therapeutic intervention. AB - Human papillomaviruses (HPV) are the aetiological agents of several types of anogenital tumours, particularly cervical carcinoma. Recent evidence also suggests a role for HPV in the development of squamous cell carcinomas of the skin, especially in immunocompromised individuals. HPV infection also produces a number of non-malignant, but nonetheless cosmetically unpleasant lesions. Therefore, any effective therapeutic treatment for HPV-induced diseases would be extremely beneficial both on humanitarian grounds as well as being economically very attractive. In this review, we will discuss the functions of the viral proteins that appear to be the most appropriate for the development of therapeutics aimed at the treatment of viral infection and virus-induced cancers. PMID- 15535405 TI - Management of antiviral resistance in patients with chronic hepatitis B. AB - A meeting of physicians and scientists involved in the management of chronic hepatitis B (CHB) was held to review current scientific data regarding antiviral resistance in hepatitis B virus (HBV) infection. The goals of the meeting were to describe current treatments for CHB, discuss emerging issues in HBV drug resistance and to delineate patient monitoring, including markers for resistance, during administration of antiviral therapy. The aim of this review article is to provide treating physicians with a framework for the management of CHB in the context of antiviral resistance. Definitions of primary and secondary antiviral treatment failure can be used to aid monitoring and early diagnosis of drug resistance before disease progression occurs as a consequence of viral breakthrough. Primary antiviral treatment failure is defined as failure of a drug to reduce HBV DNA levels by > or = 1 x log10 IU/ml within 3 months following initiation of therapy, and secondary antiviral treatment failure as a rebound of HBV replication of > or = 1 x log10 IU/ml from nadir in patients with an initial antiviral treatment effect (> or = 1 x log10 IU/ml decrease in serum HBV DNA). Confirmation of antiviral drug failure can be established by sequencing the HBV DNA polymerase and identifying specific genetic markers of antiviral drug resistance. In addition to virological assays, HBV resistance can be assessed from a clinical perspective including increased serum alanine aminotransferase levels and the development of systemic symptoms or signs of liver failure. Potential strategies to prevent the emergence of resistance and how to manage drug-resistant HBV once it emerges are discussed. PMID- 15535406 TI - Current patterns in the epidemiology of primary HIV drug resistance in North America and Europe. AB - Despite numerous studies in recent years, it is still difficult to draw general conclusions about the extent to which drug-resistant HIV-1 is transmitted. In addition to the highly stratified nature of primary resistance itself, true epidemiological surveillance has been rare and studies to date have suffered from wide variability in their designs, definitions and datasets. In the absence of consensus standards, this has resulted in a large number of isolated 'snapshots' with little scope for data-pooling and comparison. This brief review examines some of the major confounding factors that restrict the utility of individual studies and prevent the combination of studies to increase statistical power. Despite these limitations, data from North America and Europe lead to the tentative conclusion that transmission rates in these areas have generally fallen or remained stable in the past 2-3 years. However, data for the UK seem to indicate an ongoing rise in the transmission of drug resistance mutations, currently present in up to 20% of new infections. Transmission of resistant HIV represents a clinically important phenomenon, although the scale and relevance are being obscured by methodological variations and non-clinical definitions of resistance. Those of us with an interest in the epidemiology of drug resistance, whether in primary transmission or on-treatment, must learn to speak the same language if we are to establish meaningful correlations between survey datasets and the HIV-infected population as a whole. PMID- 15535407 TI - Antivirogram or phenosense: a comparison of their reproducibility and an analysis of their correlation. AB - The Antivirogram and PhenoSense assays are widely used phenotypic tests for HIV drug resistance. There are limited data on the reproducibility of each assay, and little is known about the correlation between the two. Using data from the Stanford HIV drug resistance database, we performed a comprehensive analysis of the reproducibility of each assay, and calculated the correlation and concordance of the two assays using both general IC50 fold change cutoff values and drug specific cutoff values. Although the within-assay correlations were high (rank correlation coefficients r=0.94 and r=0.95 for the Antivirogram and PhenoSense assays, respectively), the between-assay correlation was considerably lower (r=0.36). Using drug-specific cutoff values for viruses classified as resistant by the Antivirogram or PhenoSense assays, respectively, only 71.4% [95% confidence intervals (95% CI): 58.7-82.1%] and 57.0% (95% CI: 45.3-68.1%) of the samples were classified as resistant using the other assay. The poor agreement between the assays was primarily due to the extremely poor correlation between these assays for samples with low resistance values (r=0.02 and r=0.61 for samples with the Antivirogram measurements lower or higher than 2.0, respectively). Since the cutoff values for both assays are relatively low, our analysis suggests that one should be very careful when interpreting measurements that are near the cutoff values for drug resistance. PMID- 15535408 TI - Initiating highly active antiretroviral therapy and continuity of HIV care: the impact of incarceration and prison release on adherence and HIV treatment outcomes. AB - OBJECTIVE: To examine the effect of incarceration within 12 months of initiating highly active antiretroviral therapy (HAART) on non-adherence and HIV-1 RNA suppression. METHODS: We compared the adherence and virological outcomes among participants in a population-based HIV/AIDS Drug Treatment Program in British Columbia, Canada, by history of incarceration in a provincial prison. Participants who were HIV-infected, naive to HAART and who were prescribed treatment between 1 July 1997 and 1 March 2002 were eligible for this study. Logistic regression was used to determine the factors associated with non adherence and Cox proportional hazards modelling was used to determine the factors associated with HIV-1 RNA suppression adjusting for age, gender, history of drug use, baseline HIV-1 RNA, baseline CD4 cell count, type of antiretroviral regimen [two nucleosides + protease inhibitor (PI) vs two nucleosides + non nucleoside reverse transcriptase inhibitor (NNRTI)], physician's HIV-related experience for each subject and adherence as measured by pharmacy refill compliance. RESULTS: There were 1746 subjects (101 incarcerated/1645 non incarcerated) who started antiretroviral therapy between 1 July 1997 and 1 March 2002. Of those incarcerated, 50 initiated HAART while in prison and 27 subjects were released but returned to prison in the follow-up period. Subjects received antiretroviral therapy while incarcerated for a median number of 4 months [interquartile range (IQR): 2-10]. Multiple logistic regression results showed that a history of incarceration within 12 months of initiating HAART independently increased the odds of non-adherence [adjusted odds ratio (AOR): 2.40; 95% confidence interval (95% CI): 1.54-3.75]. A history of injected drug use was also associated with non-adherence (AOR: 1.49; 95% CI: 1.17-1.90). The following factors were negatively associated with non-adherence: older age (AOR: 0.81; 95% CI: 0.72-0.91), male sex (AOR: 0.50; 95% CI: 0.38-0.65) and higher physician HIV-related experience (AOR: 0.97; 95% CI: 0.96-0.98). In addition, a history of incarceration within 12 months of initiating HAART reduced the odds of achieving HIV-1 RNA suppression [adjusted hazards ratio (AHR): 0.68; 95% CI: 0.51 0.89]. Other factors negatively associated with viral suppression included a history of drug injection (AHR: 0.79; 95% CI: 0.69-0.91), two nucleosides + PI vs two nucleosides + NNRTI (AHR: 0.77; 95% CI: 0.69-0.87), higher baseline HIV-1 RNA (AHR: 0.66; 95% CI: 0.62-0.70). Higher adherence was positively associated with viral suppression (AHR: 1.38; 95% CI: 1.34-1.42). Among the 101 subjects who were incarcerated in the first year of starting HAART, the time spent in jail was positively associated with HIV-1 RNA suppression (HR: 1.06; 95% CI: 1.02-1.10). CONCLUSION: HIV-infected subjects with a history of incarceration within 12 months of initiating HAART have higher odds of non-adherence and, consequently, lower probability of achieving HIV-1 RNA suppression. The longer their sentence, however, the higher the probability of virological suppression. The British Columbian provincial prison system provided a structured setting for HAART but subjects are unable to continue this level of adherence upon release. Strategies to ensure continuation of HIV/AIDS care for HIV-infected individuals leaving the criminal justice system must be a public health priority. PMID- 15535409 TI - Quality of life in patients treated with first-line antiretroviral therapy containing nevirapine and/or efavirenz. AB - OBJECTIVE: To assess whether differences in safety profiles between nevirapine (NVP) and efavirenz (EFV), as observed in the 2NN study, translated into differences in 'health related quality of life' (HRQoL). DESIGN: A sub-study of the 2NN study, with antiretroviral-naive patients randomly allocated to NVP (once or twice daily), EFV or NVP+EFV, in addition to stavudine and lamivudine. METHODS: Comparing differences in changes of HRQoL over 48 weeks as measured with the Medical Outcomes Study HIV Health Survey (MOS-HIV) questionnaire, using analysis of variance. RESULTS: The 2NN study enrolled 1216 patients. No validated questionnaires were available for 244 patients, and 55 patients had no HRQoL data at all, leaving 917 patients eligible for this sub-study. A total of 471 (51%) had HRQoL measurements both at baseline and week 48. The majority (69%) of patients without HRQoL measurements did, however, complete the study. The change in the physical health score (PHS) was 3.9 for NVP, 3.4 for EFV and 2.4 for NVP+EFV (P=0.712). For the mental health score (MHS) these values were 6.1, 7.0 and 3.9, respectively (P=0.098). A baseline plasma HIV-1 RNA concentration (pVL) > or = 100,000 copies/ml and a decline in pVL (per log10) were independently associated with an increase of PHS. An increase of MHS was only associated with pVL decline. Patients experiencing an adverse event during follow-up had a comparable change in PHS but a significantly smaller change in MHS, compared with those without an adverse event. CONCLUSIONS: First-line ART containing NVP and/or EFV leads to an improvement in HRQoL. The gain in HRQoL was similar for NVP and EFV, but slightly lower for the combination of these drugs. PMID- 15535410 TI - Effect of long-term lamivudine in chronic hepatitis B virus-infected children. AB - OBJECTIVE: To evaluate, retrospectively, biochemical, serological and histological responses in chronic hepatitis B (CHB)-infected children who received combination therapy and continued with prolonged treatment with lamivudine (3TC). PATIENTS AND METHODS: CHB infection was defined as the presence of hepatitis B surface antigen (HBsAg), hepatitis Be antigen (HBeAg) and hepatitis B virus (HBV) DNA in serum screened at 3-month intervals for at least 1 year, serum alanine aminotransferase (ALT) levels >1.5 times the normal limit and CHB with histological activity index (HAI) >5 by liver biopsy. A total of 99 children with CHB infection were treated with IFN-alpha (three times a week, 5 MU/m2) and 3TC (4 mg/kg/d) orally for 6 months. End of therapy response (CR) was defined as ALT normalization, HBV-DNA clearance and e seroconversion. Partial responders (PR) were defined as patients who had ALT normalization and HBV-DNA clearance, but who had not had e seroconversion. Forty-five children with PR at the end of the sixth month continued to receive 3TC alone thereafter. Breakthrough infection was determined as re-emergence of HBV DNA in serum after its clearance. The response rate, side effects and the breakthrough infection rate were examined on prolonged 3TC treatment. Liver biopsy was held in 29 patients at median 32 (14-66) months of 3TC; pre- and post-treatment liver histology was compared. RESULTS: Pre- and post-treatment evaluation was carried out in 45 children [mean age: 11+/-4.2 years, 31 males (69%), 14 females (31%)] with PR at the end of the sixth month of combination therapy. The initial mean ALT values and HAI scores were 75.6+/-60 IU/l and 8+/-3.3, respectively. 3TC was continued for median 33 (14-66) months and CR was achieved in 15.6% (7/45) and 5.6% (2/36) at the end of first and second year, and 0% (none) at the end of third and fourth year, respectively. Breakthrough incidence was detected in six (13.3%) cases at 12 months and increased to 69.4% (n=25) and 82.4% (n=14) at the end of the second and third years, respectively. Patients with breakthrough continued to receive 3TC. Seroconversion and CR of the mutant virus was achieved in one patient (2.9%) at month 46 of treatment with 3TC. Liver biopsy was held in 29 cases at median 32 (14-66) months of 3TC. Pre- and post-treatment mean HAI scores were 8+/-3.3 and 3.9+/-2.1, respectively (P=0.000). Mean necrosing scores were not different at the beginning and end of therapy (P=1.0). Inflammation, bridging and fibrosis scores decreased to 0.8+/-0.6, 1.3+/-1.2 and 0.6+/-0.8, respectively (P=0.000, P=0.002, P=0.000). CONCLUSION: The long-term 3TC usage in children with PR does not induce complete response and is associated with high breakthrough incidence. However, histological improvement is achieved and/or sustained even in children with HBV DNA breakthrough. PMID- 15535411 TI - Superior therapeutic efficacy of alphavirus-mediated immunization against human papilloma virus type 16 antigens in a murine tumour model: effects of the route of immunization. AB - In our efforts to develop a strong, effective immune response against cervical carcinoma and premalignant disease, we study the use of recombinant Semliki Forest virus (SFV) encoding the oncoproteins E6 and E7 from high-risk human papilloma viruses (HPVs). Optimal immunization conditions are required for immunotherapeutic treatment of cervical cancer as it has been postulated that cervical cancer patients are immune-suppressed and/or immunologically tolerant for HPV. We previously generated an optimized construct encoding a fusion protein of HPV16 E6 and E7 and a translational enhancer (enhE6,7). Immunization of mice with SFV-enhE6,7 was shown to induce cytoxic T cell (CTL) responses and resulted in the eradication of established tumours. We now demonstrate, using HPV16 specific MHC class I tetramers, that high pCTL frequencies can be induced. However, this induction is strongly influenced by the route of immunization applied. Whilst in bulk CTL assays, requiring in vitro restimulation, CTL activity can be observed upon s.c., i.p., i.v. and i.m. immunization, detectable pCTL frequencies, without in vitro restimulation, are only induced upon i.m. and i.v. immunization. The route of immunization also strongly influences the dose of viral vector needed to induce CTLs and tumour therapy. As few as 5x10(4) SFV enhE6,7, primed and boosted i.v., are needed to eradicate tumours in six out of seven mice treated. Furthermore, exponentially growing tumours of approximately 500 mm3 in size were seen to completely resolve and even tumours as large as 1500 mm3 decreased to one-third of their size. Apart from this potency, SFV vectors can safely be used for the expression of oncoproteins such as E6 and E7, since the viral RNA is not integrated in the host genome. Thus SFV-enhE6,7 meets with the criteria that a vaccine against cervical cancer should be safe and induce a very strong, long-lasting CTL response, strong enough to eradicate existing tumours. PMID- 15535412 TI - Variability in the interpretation of transmitted genotypic HIV-1 drug resistance and prediction of virological outcomes of the initial HAART by distinct systems. AB - High level HIV-1 drug resistance in recently infected treatment-naive individuals correlates with sub-optimal virological responses to highly active antiretroviral therapy (HAART). To determine whether genotypic HIV-1 drug resistance in chronic naive patients, as interpreted by various systems, could predict the virological outcomes of HAART, isolates from patients enrolled in a prospective observational cohort (ICoNA) prior to treatment start were genotyped. Genotypic susceptibility scores (GSS) assigned to the initial HAART regimens using the interpretations of pre-therapy resistance mutations by 13 systems were related to virological outcomes. Of 415 patients, 42 (10%) had at least one major resistance mutation. According to the different interpretations, 1.9-20.5% of patients had some level of resistance to at least one drug in the initial regimen. In multivariable analysis, GSS from two systems significantly predicted the time to virological success: Rega 5.5, for each unit increase in GSS adjusted relative hazard (RH) 1.86 [95% confidence intervals (95% CI): 1.15-3.02] and hivresistanceWeb v3, RH 1.87 (95% CI: 1.00-3.48). With three other systems, GSS showed a trend towards a significant prediction of success: Retrogram 1.6, RH 2.33 (95% CI: 0.98-5.53), Menendez 2002, RH 2.36 (95% CI: 0.97-5.72) and Stanford hivdb, RH 2.06 (95% CI: 0.94-4.49). Genotypic resistance testing coupled with adequate interpretation in chronic naive patients can usefully identify those at risk of sub-optimal virological response to HAART. This work was presented in part at the First European Workshop on HIV Drug Resistance. Luxembourg, 4-6 March 2003 (Abstract 44); and at the 9th European Conference on CIinical AIDS Therapy. Warsaw, 25-28 October 2003 (Abstract F6/7). PMID- 15535415 TI - Comparison of dual-energy X-ray absorptiometry machines for measuring fat distribution changes of HIV-associated lipodystrophy. AB - BACKGROUND: Dual-energy X-ray absorptiometry (DXA) is regarded as a useful objective method for diagnosis of HIV-associated lipodystrophy (LD). OBJECTIVE: To determine whether the use of different DXA machines may affect the diagnosis of LD. METHODS: Body composition in 24 HIV-infected patients, 12 of whom had clinically diagnosed LD, was measured on two DXA machines (Lunar and Hologic). RESULTS: Hologic gave significantly higher values than Lunar DXA for total body fat percent (22.9 vs 20.5%, P<0.001), arm fat percent (27.9 vs 14.5%, P<0.001) and leg fat percent (23.4 vs 17.5%, P<0.001), and significantly lower values than Lunar for trunk fat percent (22.3 vs 24.7%, P<0.001) and trunk-to-limb fat percent ratio (0.89 vs 1.62, P<0.001). When measured by Lunar, patients with LD had significantly lower leg fat percent (13.5 vs 21.6%, P=0.04) and higher trunk to-limb fat percent ratio (1.85 vs 1.38; P=0.012) than those without LD. When measured by Hologic, patients with LD had values of leg fat percent (20.1 vs 26.8%, P=0.11) and trunk-to-limb fat percent ratio (0.95 vs 0.85, P=0.43) that did not differ significantly from those without LD. There was a significant difference in the contribution to a composite LD score between Lunar and Hologic measurements. CONCLUSION: DXA machines from different manufacturers give major differences in measurements of body fat content and distribution, and this may affect the ability to distinguish patients with LD from those without LD. Further standardization of DXA technology is needed before widespread application in the clinical diagnosis of LD. PMID- 15535413 TI - The correlation between plasma concentrations of protease inhibitors, medication adherence and virological outcome in HIV-infected patients. AB - BACKGROUND: Although adherence clearly influences response to antiretroviral therapy (ART), accurate assessment of adherence is problematic. The objective of this analysis was to assess the independent predictive value of protease inhibitor (PI) concentrations as a supplement to self-report as markers of medication adherence. METHODS: This retrospective analysis was conducted from a prospective clinical trial designed to compare the outcomes of frequent versus infrequent HIV RNA measurement used to manage antiretroviral therapy. For 131 patients, self-reported medication adherence, HIV RNA levels, CD4 counts and PI concentrations (unannounced, random samples) were measured at baseline (when patients changed to a new regimen) and every 2 months thereafter. The change in HIV RNA from baseline to month 6 (area-based measure) was used to evaluate overall response. The proportion of measured PI concentrations below the detection limit was used as an alternative marker of adherence. An undetectable concentration would be expected after missing a single dose. RESULTS: The mean baseline CD4 count was 125 cells/mm3 and the mean HIV RNA level was 4.7 log10 copies/ml. The mean change in log10 HIV RNA was -0.73 copies/ml. The mean percentage of self-reported adherence was 91% (range: 15-100%) and the mean proportion of undetectable PI concentrations was 27% (range: 0-100%, mean 2.5 samples/patient). The correlation between the two measures was -0.23 (P=0.009). In a multivariate model, percentage of visits with undetectable PI concentrations (P=0.02), percentage of medication adherence (P=0.02), baseline HIV RNA level (P=0.005), prior PI use (P=0.0004), prior lamivudine (3TC) use (P=0.0009) and randomization to the frequent HIV RNA measurement group (P<0.0001) were all related to change in HIV RNA. After accounting for adherence, patients who always had detectable PI concentrations had an average of 0.4 log10 additional HIV RNA reduction compared with those who had no detectable concentrations. CONCLUSIONS: Repeated, random PI concentration values are independently predictive of virological response and may add to self-report of adherence in understanding the response to ART. PMID- 15535414 TI - PKR gene expression and response to pegylated interferon plus ribavirin therapy in chronic hepatitis C. AB - Pegylated interferon (PEG-IFN) alpha combined with ribavirin is the current standard treatment for hepatitis C, but around 50% of patients do not respond for reasons that are not fully understood. To explore the regulation of IFN-inducible protein kinase (PKR), we have measured PKR mRNA levels in peripheral blood mononuclear cells (PBMCs) and in liver biopsies from patients with chronic hepatitis C. PBMCs were also analysed after in vitro incubation with IFN and during antiviral therapy. Non-responders to PEG-IFN plus ribavirin had pre treatment PKR mRNA levels in PBMCs (0.1+/-0.0074) and in liver (0.102+/-0.051) that were significantly higher than those of responders (PBMCs: 0.023+/-0.014, P=0.0005; liver: 0.034+/-0.020; P=0.0002). On the other hand, PKR mRNA levels in PBMCs were similar in non-responders and in responders after in vitro exposure to IFN (0.434+/-0.301 vs 0.403+/-0.222; P=NS) and during therapy (0.31+/-0.10 vs 0.30+/-0.12; P=NS). These results indicate that in hepatitis C, non responsiveness to IFN-alpha is associated with pre-treatment up-regulation of the PKR gene, evidence that the infecting hepatitis C virus is able to stimulate endogenous IFN production, being resistant to its antiviral effect. On the other hand, the PKR gene response to exogenous IFN was similar in responders and non responders, at least in PBMCs, suggesting that variations in its activation are not major determinants of the outcome of antiviral treatment. PMID- 15535416 TI - The plasma and intracellular steady-state pharmacokinetics of lopinavir/ritonavir in HIV-1-infected patients. AB - Therapeutic drug monitoring of protease inhibitors (PIs) is usually performed on plasma samples although their antiretroviral effect takes place inside cells. Little is known, however, about the intracellular accumulation and related plasma pharmacokinetics of PIs such as lopinavir/ritonavir (LPV/RTV). Therefore, we studied the plasma and intracellular (cell-associated) steady-state pharmacokinetics of this PI combination in a dosage of 400/100 mg twice daily in a non-randomized cohort of HIV-1-infected individuals. Plasma (0-12 h) and peripheral blood mononuclear cell (PBMC; 0-8 h) samples were drawn during a 12-h dosing interval in 11 subjects. The plasma concentrations versus time curves of LPV and RTV were characterized by an irregular absorption phase showing double peaks (Cmax) in most subjects and single-peaks in the remaining patients between 1 and 3 h after drug intake. Pre-dose concentrations of both agents in plasma were significantly higher than the concentrations at the end of the dosing interval indicating the presence of a circadian rhythm in their pharmacokinetics. The course of the intracellular concentrations versus time curves appeared to be similar to the plasma concentration curves, with the highest intracellular concentration measured 3 h after drug intake. The intracellular RTV concentrations were higher than reported in vitro EC50 values and might therefore contribute to the antiretroviral effect of LPV/RTV. The median intracellular-to plasma concentration ratios (interquartile range) were 1.18 (0.74-2.06) and 4.59 (3.20-7.70) for LPV and RTV, respectively. In conclusion, both LPV and RTV accumulate to potential therapeutic concentrations in PBMCs. Irregular absorption and circadian plasma clearance patterns were observed for the PI combination LPV/RTV. PMID- 15535417 TI - Baseline resistance and virological outcome in patients with virological failure who start a regimen containing abacavir: EuroSIDA study. AB - OBJECTIVES: To investigate the ability of several HIV-1 drug-resistance interpretation systems, as well as the number of pre-specified combinations of abacavir-related mutations, to predict virological response to abacavir containing regimens in antiretroviral therapy-experienced, abacavir-naive patients starting an abacavir-containing regimen in the EuroSIDA cohort. PATIENTS AND METHODS: A total of 100 HIV-infected patients with viral load (VL) >500 copies/ml who had a plasma sample available at the time of starting abacavir (baseline) were included. Resistance to abacavir was interpreted by using eight different commonly used systems that consisted of rules-based algorithms or tables of mutations. Correlation between baseline abacavir-resistance mutations and month 6 virological response was performed on this population using a multivariable linear regression model accounting for censored data. RESULTS: The baseline VL was 4.36 log10 RNA copies/ml [interquartile range (IQR): 3.65-4.99 log10 RNA copies/ml] and the median CD4 cell count was 210 cells/microl (IQR: 67 305 cells/microl). Our patients were pre-exposed to a median of seven antiretrovirals (2-12) before starting abacavir therapy. The median (range) number of abacavir mutations (according to the International AIDS Society-USA) detected at baseline was 3.5 (0-8). Overall, the Kaplan-Meier estimate of the median month 6 VL decline was 0.86 log10 RNA copies/ml [95% confidence intervals (95% CI): 0.45-1.24]. The VL in those patients (n=31) who intensified treatment by adding only abacavir decreased by a median 0.20 log10 RNA copies/ml (95% CI: 0.18; +0.94). The proportion of patients who harboured viruses fully resistant to abacavir among the eight genotypic resistance interpretation algorithms ranged from 12% [Agence Nationale de Recherches sur le SIDA (ANRS)] to 79% [Stanford HIV RT and PR Sequence Database (HIVdb)]. Some interpretation systems showed statistically significant associations between the predicted resistance status and the virological response while others showed no consistent association. The number of active drugs in the regimen was associated with greater virological suppression (additional month 6 VL reduction per additional sensitive drug=0.51, 95% CI: 0.15-0.88, P=0.006); baseline VL was also weakly associated (additional month 6 VL reduction per log10 higher=0.30, 95% CI: -0.02; +0.62, P=0.06). In contrast, the number of drugs previously received was associated with diminished viral reduction (additional month 6 VL reduction per additional drug=-0.14, 95% CI: -0.28; 0.00, P=0.05). CONCLUSIONS: Our results revealed a high degree of variability among several genotypic resistance interpretation algorithms currently in use for abacavir. Therefore, the interpretation of genotypic resistance for predicting response to regimens containing abacavir remains a major challenge. PMID- 15535418 TI - Effects of zidovudine, stavudine and beta-aminoisobutyric acid on lipid homeostasis in mice: possible role in human fat wasting. AB - OBJECTIVE: Although mitochondrial DNA (mtDNA) depletion could play a role in nucleoside reverse transcriptase inhibitor-induced lipoatrophy, poor correlations between fat mtDNA levels and lipoatrophy suggest additional mechanism(s). Stavudine (d4T), zidovudine (AZT) and the thymine catabolite, beta aminoisobutyric acid (BAIBA), but not zalcitabine (ddC) or didanosine (ddI), can increase fatty acid oxidation in liver mitochondria and plasma ketone bodies in mice. Since fat oxidation in non-adipose tissue can influence body adiposity, we sought to determine whether d4T, AZT and BAIBA can cause lipoatrophy in mice by this catabolic mechanism. METHODS: Lean or obese ob/ob mice were treated for 6 weeks with d4T, AZT or BAIBA, and lean mice with ddC or ddI. Body fat mass was assessed by dual energy X-ray absorptiometry, and mtDNA by Slot blot hybridization in epididymal fat. RESULTS: Whereas ddC or ddI did not change plasma beta-hydroxybutyrate and body fat mass, d4T, AZT and BAIBA increased plasma beta-hydroxybutyrate in lean mice suggesting increased hepatic fatty acid oxidation and ketogenesis. Despite unchanged food consumption, a supra pharmacological dose of d4T tended to decrease, whilst AZT and BAIBA decreased body fat mass. Fat mtDNA and plasma triglycerides, cholesterol, glucose, insulin, leptin and adiponectin levels were unchanged. In obese mice, d4T, AZT and BAIBA did not increase plasma beta-hydroxybutyrate, and only AZT decreased body fat mass without reducing fat mtDNA. CONCLUSIONS: d4T and AZT can enhance hepatic fat oxidation and cause fat wasting, without decreasing adipose tissue mtDNA and without causing insulin resistance in mice. BAIBA, a thymine catabolite, reproduces these effects. These catabolic effects could play a role in the lipoatrophy, which can occur in AZT- or d4T-treated patients. PMID- 15535419 TI - The management of hepatitis B virus/HIV-1 co-infected patients starting their first HAART regimen. Treating two infections for the price of one drug? AB - We examined the impact of a lamivudine-containing highly active antiretroviral therapy (HAART) regimen on 164 hepatitis B virus/HIV co-infected individuals starting their first HAART. Lamivudine-treated patients (accounting for 73% of the study population) showed a significantly lower level of alanine aminotransferase over follow-up [-81.1 mU/ml mean difference; 95% confidence intervals (95% CI): -30.3; -131.7, P=0.003] and a significantly reduced risk of liver-related morbidity/mortality [Relative hazard (RH)=0.07; 95% CI: 0.01-0.38, P=0.002] than those starting a lamivudine sparing-regimen. PMID- 15535420 TI - Expression of pregnane-X-receptor transcript in peripheral blood mononuclear cells and correlation with MDR1 mRNA. AB - P-glycoprotein (P-gp) limits bioavailability and accumulation of HIV protease inhibitors (PIs). PIs are ligands for the pregnane-X-receptor (PXR), which regulates P-gp expression. This occurs when ligands activate the receptor, initiating binding to response elements in the MDR1 promoter. PXR also activates cytochrome P4503A4 (CYP3A4) and a correlation between hepatic PXR and CYP3A4 mRNA has been reported. We have examined the relationship between MDR1 and PXR mRNA in peripheral blood cells and demonstrate a significant correlation in 18 volunteers (R2=0.4; P<0.005). PXR was approximately 250-fold lower in peripheral blood mononuclear cells than in liver (1.6+/-1.2 vs 450+/-298; n=6; P<0.01). PMID- 15535421 TI - Cidofovir on HHV-8 in BCBL-1 cells. PMID- 15535422 TI - Increasing prevalence of HIV-1 reverse transcriptase mutation K65R correlates with tenofovir utilization. PMID- 15535423 TI - Testosterone/progestin regimens: a realistic option for male contraception? AB - Exogenously administered testosterone functions as a contraceptive in the male by suppressing the secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) from the pituitary. After 2 to 3 months of treatment, low levels of FSH and LH lead to markedly decreased sperm concentrations. This approach to contraceptive development appears safe and fully reversible, however, sperm concentrations are not suppressed to zero in all men. Therefore, researchers have combined testosterone with progestins to further suppress pituitary gonadotropins and optimize contraceptive efficacy. Current testosterone/progestin combinations using newer, long-acting forms of testosterone combined with the progestins such as desogestrel, etonorgestrel, norethisterone, medroxyprogesterone and levonorgestrel suppress spermatogenesis-to azoospermia without causing severe side effects in close to 90% of men. Significant suppression of spermatogenesis is observed in the remainder of individuals. These findings may soon result in the marketing of a safe, reversible and effective hormonal contraceptive for men. PMID- 15535424 TI - Use of estrogen antagonists and aromatase inhibitors in breast cancer and hormonally sensitive tumors of the uterine body. AB - The new anti-estrogens, including the selective estrogen receptor modulators (e.g., toremifene, droloxifene, idoxifene, raloxifene and arzoxifene), the selective estrogen receptor downregulators (e.g., fulvestrant), and the new steroidal (e.g., exemestane) and non-steroidal (anastrozole and letrozole) aromatase inhibitors have been investigated in the treatment of breast cancer and hormonally sensitive tumors of the uterine body. In postmenopausal women, anastrozole, letrozole and exemestane appear to have superior efficacy and an improved toxicity profile than tamoxifen, and fulvestrant seems to be at least as effective as anastrozole in patients in whom tumors have progressed after prior endocrine treatment. Preliminary data showed a high response rate and a favorable toxicity profile for arzoxifene in endometrial cancer, whereas letrozole could represent a new interesting therapeutic tool for endometrial cancer as well as for low-grade endometrial stromal sarcoma. PMID- 15535425 TI - Peroxisome proliferator activated receptor ligands for the treatment of insulin resistance. AB - Peroxisome proliferator activated receptors (PPARs) are a group of ligand activated transcription factors that play critical roles in the regulation of energy metabolism. Synthetic agonists for these receptors are used in the treatment of type 2 diabetes mellitus and dyslipidemia. Current data linking PPAR and/or PPAR ligands with the treatment of insulin resistance will be critically reviewed, with the focus on important advances over the past year. PMID- 15535426 TI - G protein-coupled receptors as therapeutic targets for obesity and type 2 diabetes. AB - The prevalence of obesity and type 2 diabetes, two strongly correlated disorders, is increasing worldwide. Weight loss can reduce the risk of developing type 2 diabetes and the pharmacological treatments normally required to manage this disorder. Even though dietary and lifestyle changes may eventually reduce obesity for some individuals, new safe and more efficacious drugs are required for successful weight reduction and treatment of type 2 diabetes in a large proportion of obese individuals. In addition to targeting known G protein-coupled receptors (GPCRs), several orphan GPCRs expressed in central nervous system areas known to regulate feeding may provide new targets for the treatment of obesity. Similarly, the pancreas contains numerous islet GPCRs as well as an abundance of orphan GPCRs that potentially could emerge as targets for future antidiabetic compounds. One of the major challenges facing the pharmaceutical industry is how to rapidly establish the function and therapeutic relevance of orphan GPCRs, some of which may represent novel targets for the discovery of the next generation of drugs to effectively treat obesity and type 2 diabetes. This review will focus on the significant potential of known and orphan GPCRs as targets for the discovery of new drugs to successfully treat these serious disorders. PMID- 15535427 TI - Melanocortin-4 receptor agonists for the treatment of obesity. AB - The identification of a highly efficacious anti-obesity agent remains an illusive goal. While many avenues of investigation have been pursued, none of the existing compounds claim much more than a 10% reduction in weight in humans (over a one year period with diet and exercise). Nonetheless, the potential reward for fulfilling this unmet medical need has kept interest levels high in the research community. The recent explosion of genetic information has identified numerous potential targets for drug screening. The melanocortin-4 receptor is a promising target and is currently being intensively investigated by many companies and academic research groups. PMID- 15535428 TI - Insights from growth hormone receptor blockade. AB - Pegvisomant is a genetically manipulated growth hormone (GH) that disables signal transduction through the GH receptor and thus functions as a GH receptor antagonist. In a series of studies of the metabolic effects of GH in healthy male individuals, pegvisomant has been used to create a model of acute GH deficiency but without the typical alterations in body composition. This review summarizes studies of the effects of GH receptor blockade and fasting, either alone or in combination, on determinants of GH release. Based on these and other data we present a model to explain the somatotroph hyperactivity of fasting PMID- 15535429 TI - Calcimimetics and hyperparathyroidism. AB - The calcium-sensing receptor (CaSR) is a key regulator of parathyroid hormone (PTH) secretion and parathyroid proliferation. This review discusses the role of the CaSR in hyperparathyroidism (HPT), and the exploitation of the CaSR as a therapeutic target for the treatment of HPT. Calcimimetics act as CaSR agonists or allosteric activators and thereby potentiate the effects of extracellular Ca2+ on parathyroid cell function. Results from clinical trials using calcimimetics to treat primary and secondary HPT, as well as primary HPT from parathyroid carcinoma, suggest that calcimimetic compounds could provide an effective alternative or additional therapeutic approach for various forms of HPT. PMID- 15535430 TI - Bazedoxifene (Wyeth). AB - Bazedoxifene is a tissue-specific selective estrogen receptor modulator being developed by Wyeth Pharmaceuticals (formerly Wyeth-Ayerst Laboratories) to be used alone for the prevention and treatment of osteoporosis in postmenopausal women and in combination with Premarin for menopausal symptoms. As of March and June 2004, worldwide phase III trials for bazedoxifene, and bazedoxifene in combination with Premarin, were ongoing. PMID- 15535431 TI - LAF-237 (Novartis). AB - Novartis is developing LAF-237, one of a series of orally active dipeptidylpeptidase IV inhibitors, as a potential once daily therapeutic agent for type 2 diabetes. Phase III trials began in the first quarter of 2004. PMID- 15535432 TI - alpha-glucosidase (CHO) (Genzyme). AB - Genzyme General is developing recombinant human alpha-glucosidase, produced in mammalian cell culture, as a potential treatment for Pompe disease. By July 2004, enrollment was completed in two clinical trials and an observational study in adults. Genzyme was planning to file for regulatory approval in Europe during 2004, followed by filings in the US and Japan in mid-2005. PMID- 15535433 TI - Aryplase (Biomarin). AB - BioMarin is developing Aryplase (BM-102), a recombinant form of the enzyme N acetylgalactosamine 4-sulfatase, for the potential treatment of mucopolysaccharidosis type VI (Maroteaux-Lamy Syndrome). By November 2003 enrollment for a pivoltal phase III trial was complete. PMID- 15535434 TI - Fourth International Conference on Innovations and Challenges in Prostate Cancer: Prevention, Detection and Treatment. PMID- 15535435 TI - The role of inflammation in the pathogenesis of prostate cancer. AB - PURPOSE: A new hypothesis for the etiology of prostate cancer is that chronic or recurrent prostate inflammation may initiate and promote prostate cancer development. MATERIALS AND METHODS: We reviewed the current direct and indirect evidence from epidemiology, genetics, molecular biology and histopathology implicating inflammation in the pathogenesis of prostate cancer. RESULTS: The case for prostate inflammation as a cause of prostate cancer is compelling. Epidemiology data have correlated prostatitis and sexually transmitted infections with increased prostate cancer risk and intake of anti-inflammatory drugs and antioxidants with decreased prostate cancer risk. Genetic studies have identified RNASEL, encoding an interferon inducible ribonuclease, and MSR1, encoding subunits of the macrophage scavenger receptor, as candidate inherited susceptibility genes for familial prostate cancer. Somatic silencing of GSTP1, encoding a glutathione S-transferase capable of defending against oxidant cell and genome damage, has been found in almost all prostate cancer cases. Proliferative inflammatory atrophy lesions containing activated inflammatory cells and proliferating epithelial cells appear likely to be precursors to prostatic intraepithelial neoplasia lesions and prostatic carcinomas. CONCLUSIONS: Emerging hints that prostate inflammation may contribute to prostatic carcinogenesis will provide opportunities for the discovery and development of new drugs and strategies for prostate cancer prevention. PMID- 15535436 TI - The relative impact and future burden of prostate cancer in the United States. AB - PURPOSE: In the last 2 decades there has been increasing awareness of and research on prostate cancer. We considered prostate cancer in the context of other major causes of morbidity and mortality. MATERIALS AND METHODS: Surveillance, Epidemiology, and End Results, and other government databases were used to 1) compare the incidence and mortality burden of prostate cancer with that of other cancers and major causes of death; 2) examine the changing trends in prostate cancer mortality using joinpoint analysis and 3) consider the future burden of prostate cancer in the United States due to aging. To project the future burden of prostate cancer year 2000 mortality rates were applied to age specific population projections from the United States Census Bureau. RESULTS: Prostate cancer continues to have the highest incidence rate and the second highest mortality rate of any cancer in American men and it ranks among the top 10 overall causes of death. The mortality rate of prostate cancer has been decreasing steadily at approximately 4% yearly since 1994, while the incidence rate has been increasing at slightly less than 2% yearly since 1995. CONCLUSIONS: The overall decrease in the mortality rate of prostate cancer since 1994 may be due to improvements in screening, diagnosis and treatments. Despite the decreasing mortality rate the aging of the American population and the exponential increase in prostate cancer incidence and mortality rates by age are important to consider when assessing the future morbidity and mortality burden of this disease and the appropriate allocation of health care resources. PMID- 15535437 TI - Using molecular markers to predict outcome. AB - PURPOSE: Developing molecular tests to predict prostate cancer progression requires first defining meaningful clinical end points and defining strategies to take advantage of emerging technology. MATERIALS AND METHODS: The select relevant literature was reviewed concerning clinical trials, clinical prostate cancer nomograms, molecular biomarker development and molecular prostate cancer imaging. RESULTS: There is controversy regarding the use of prostate specific antigen or biochemical failure following prostatectomy or radiation therapy for clinically localized prostate cancer as a marker of progression. As a consequence, advances in prostate cancer biomarker development may require using population based cohorts or cases from clinical trials to identify meaningful associations. Whereas the discovery of novel candidate biomarkers was slow 5 to 10 years ago and often resulted from serendipity, advances in high throughput technologies have led to the identification of a large number of candidate genes. Strategies to identify candidate genes include the use of expression array analysis, single nucleotide polymorphism arrays (single nucleotide polymorphism chips), proteomics and bioinformatics. Monitoring the progression of prostate cancer has been limited to standard approaches such as computerized tomography or magnetic resonance imaging, which in general do not delineate the extent of disease. By carefully selecting novel prostate cancer biomarkers future work should allow in vivo monitoring of prostate cancer. This will represent a revolutionary advance in our ability to monitor prostate cancer progression and ultimately it may be one of the most important applications of cancer biomarkers. CONCLUSIONS: Emerging technology should allow us to analyze clinical prostate cancer trials with sufficient followup to help develop meaningful markers of prostate cancer progression. PMID- 15535438 TI - Expression of genes and proteins specific for prostate cancer. AB - PURPOSE: We reviewed the pertinent literature and discuss high throughput approaches to the identification of molecular markers characteristic of prostate cancer. MATERIALS AND METHODS: A PubMed search was performed for expression array studies comparing prostate cancer tissues with nonmalignant tissues and serum proteomic studies comparing patients with prostate cancer and controls. RESULTS: The expression of several genes may help distinguish prostate cancer cells from nonmalignant prostatic epithelial cells. A few genes, such as those coding for hepsin, LIM protein and alpha-methylacyl-coenzyme A racemase, have consistently been identified as being over expressed in cancer cells compared with nonmalignant cells. Proteomic analysis using mass spectroscopy has identified spectral patterns that have a significant correlation with the presence of prostate cancer. The optimal presentation of the spectra may depend highly on technical factors and experimental protocols have yet to be standardized among laboratories. Moreover, to our knowledge the serum proteins that comprise the mass spectral patterns of prostate cancer have yet to be identified. CONCLUSIONS: High throughput techniques have the potential to aid in prostate cancer diagnosis, early detection and developmental therapies. Markers with the potential to aid in diagnosis have been identified and targets for imaging or therapeutic approaches appear to be at hand. Mass spectral analysis of serum is in the early stages and it requires standardization of technical parameters among laboratories. In addition, the identification of target proteins will contribute to ease of detection and cross-laboratory verification. PMID- 15535439 TI - Predicting prostate cancer behavior using transcript profiles. AB - PURPOSE: Prostate cancer represents a disease with diverse clinical outcomes. Treatment strategies that optimize benefit and minimize morbidities depend on accurate estimates of disease status and likelihood of progression. Emerging technologies capable of qualitatively and quantitatively profiling genes expressed by neoplastic tissues may provide insights into tumor behavior. This review discusses the use of microarray based transcript expression profiling to stratify human cancers into risk categories. MATERIALS AND METHODS: MEDLINE was used to perform a comprehensive literature review of reports describing the assessment of gene expression profiles in malignant diseases. Particular emphasis was placed on studies developing models using individual genes or gene cohorts as predictors of prostate cancer outcome. RESULTS: Alterations in the expression of individual genes identified by microarray analyses have been used in studies of outcome in cancers of the prostate and other tissue types. Profiles of expressed genes have been used to develop prediction models that stratify cancers into prognostic categories based on relapse rates or responses to therapy. CONCLUSIONS: Gene expression profiles offer an opportunity for acquiring molecular determinants correlating with clinical outcome. With rare exceptions these profiles have yet to be validated or used in prospective studies. Future research will benefit from assessments of intratumor heterogeneity and host factors such as the immune response and hormonal milieu. The prospective validation of predictive models will serve to prove usefulness in the clinical arena. PMID- 15535440 TI - An overview of chemotherapy trials in localized and recurrent nonmetastatic prostate cancer. AB - PURPOSE: High risk localized and recurrent prostate cancers remain difficult problems for clinicians. Definitive local treatments such as surgery and radiation therapy cure only a minority of these patients. As a result, efforts are being made to decrease the risk of recurrence and delay progression to symptomatic hormone refractory disease using chemotherapy prior to, during or after definitive local therapy. MATERIALS AND METHODS: A MEDLINE literature review was performed of studies evaluating chemotherapy and other systemic therapies for localized prostate cancer. In addition, ongoing randomized clinical trials were reviewed. RESULTS: Chemotherapy in the management of localized and recurrent disease is evolving. Early studies demonstrated the feasibility of administering chemotherapy to patients prior to and immediately after local therapy, and in the setting of an increasing prostate specific antigen state. Although chemotherapy has established activity and acceptable toxicity in patients with hormone refractory prostate cancer, more extensive use in patients with androgen dependent disease will require data from randomized studies to determine its efficacy. At least 5 ongoing, randomized, phase III trials currently are evaluating the role of chemotherapy in various prostate cancer disease states, including nonmetastatic, hormone refractory disease, the increasing prostate specific antigen state, and adjuvant and neoadjuvant settings in patients at high risk. CONCLUSIONS: Chemotherapy is being investigated actively in randomized trials of earlier states of prostate cancer. Enrollment onto clinical trials should be the highest priority for investigators and clinicians. PMID- 15535441 TI - Optimizing hormone therapy in localized prostate cancer: focus on external beam radiotherapy. AB - PURPOSE: The use of hormone therapy combined with radiotherapy (RT) and prior to radical prostatectomy was explored as part of the management of localized prostate cancer. MATERIALS AND METHODS: Recent randomized studies and ongoing clinical trials of hormone therapy were reviewed to help identify patients who will benefit from combined treatment. RESULTS: Although short-term androgen ablation improves cause specific survival in low Gleason score, bulky prostatic neoplasms, long-term androgen ablation is required for high grade prostate cancer. Whole pelvic RT may be important for maximizing the biochemical interaction between RT and androgen ablation in patients at high risk for pelvic lymph node involvement. Ongoing studies will contribute to our overall understanding of combined modality treatment. CONCLUSIONS: Androgen ablation is an important part of the management of prostate cancer when external beam RT is used. Long-term androgen ablation should be performed in bulky, high Gleason score cases, while short-term androgen ablation should be used in bulky, low Gleason score cases. Patients at intermediate risk are candidates for short-term androgen ablation but there are as yet no definitive randomized trials assessing an overall treatment benefit. Patients with low risk prostate cancer should probably not receive androgen ablation unless additional data support a substantial clinical benefit. The lack of advantage observed in patients undergoing preoperative androgen ablation compared with the advantages seen in patients who undergo androgen ablation and RT seems to indicate that at least in some situations there is an advantageous biological interaction between RT and androgen ablation. This mechanism remains to be elucidated. PMID- 15535442 TI - Prostate specific antigen doubling time as a surrogate end point for prostate cancer specific mortality following radical prostatectomy or radiation therapy. AB - PURPOSE: A short posttreatment prostate specific antigen (PSA)-doubling time (DT) following radical prostatectomy or radiation therapy was evaluated as a surrogate end point for prostate cancer specific mortality (PCSM). MATERIALS AND METHODS: Baseline, treatment and followup information was compiled on a cohort of 8,669 patients with prostate cancer treated with surgery (5,918) or radiation (2,751) from January 1, 1988 to January 1, 2002 for clinical stage T1c-4NxMo prostate cancer, forming the study cohort. Cox regression analysis was used to test whether Prentice criteria were violated in this cohort. RESULTS: After PSA defined recurrence PSA-DT less than 3 months and the specific value of PSA-DT at 3 months or greater were statistically significantly associated with time to PCSM and with time to all cause mortality after PSA defined recurrence (each Cox p <0.001). Treatment received was not statistically significant associated with time to PCSM following PSA defined recurrence in patients with PSA-DT less than 3 months (Cox p = 0.90) and in patients with PSA-DT 3 months or greater (Cox p = 0.28). Furthermore, after PSA defined recurrence PSA-DT less than 3 months was statistically significantly associated with PCSM (HR 19.6, 95% CI 12.5 to 30.9). CONCLUSIONS: Posttreatment PSA-DT appears to be a surrogate end point for PCSM following surgery or radiation therapy. We recommend that consideration should be given to enrollment onto a clinical trial and/or initiating androgen suppression therapy at the time of PSA defined recurrence when PSA-DT is less than 3 months to delay the imminent sequelae of metastatic bone disease. PMID- 15535443 TI - Active surveillance with selective delayed intervention: using natural history to guide treatment in good risk prostate cancer. AB - PURPOSE: This article reviews the data supporting an approach of active surveillance with selective delayed intervention for good risk localized prostate cancer. The challenge is to identify those patients who are not likely to experience significant progression, while offering radical therapy to those who are at risk. MATERIALS AND METHODS: A prospective phase 2 study of active surveillance with selective delayed intervention was initiated in 1995. Patients were treated initially with surveillance, while those who had a prostate specific antigen (PSA) doubling time (DT) of 2 years or less, or grade progression on re biopsy were offered radical intervention. The remainder were closely monitored. RESULTS: The cohort consisted of 299 patients with good risk prostate cancer or intermediate risk prostate cancer in men older than 70 years. Median PSA DT was 7.0 years and 35% of the men had a PSA DT of greater than 10 years. The majority of patients remain on surveillance. At 8 years overall actuarial survival was 85% and disease specific survival was 99%. CONCLUSIONS: Most men with favorable risk prostate cancer will die of unrelated causes. The approach of active surveillance with selective delayed intervention based on PSA DT represents a practical compromise between radical therapy in all, which results in overtreatment in patients with indolent disease, and watchful waiting with palliative therapy only, which results in under treatment in those with aggressive disease. Results at 8 years are favorable. Longer followup will be required to confirm the safety of this approach in men with long (greater than 15-year) life expectancy. PMID- 15535444 TI - Osteoporosis and obesity in men receiving hormone therapy for prostate cancer. AB - PURPOSE: A presentation on osteoporosis, obesity and obesity related disease at the Conference on Innovations and Challenges in Prostate Cancer: Prevention, Detection and Treatment is summarized. MATERIALS AND METHODS: A focused literature review was done. RESULTS: Gonadotropin-releasing hormone (GnRH) agonists decrease bone mineral density and increase fracture risk. GnRH agonists also increase weight and fat mass, and decrease lean body mass. Treatment related changes in body composition may contribute to fatigue and fracture risk. The phenotype of men with GnRH agonist shares some features with the insulin resistance syndrome, raising the possibility that GnRH may also increase the risk of diabetes mellitus and cardiovascular disease. CONCLUSIONS: The routine use of GnRH agonists in men with long life expectancy increases the importance of understanding and preventing the unintended adverse effects of treatment. Some adverse effects have the potential to impact not only quality of life, but also noncancer mortality. Additional research is needed to characterize better the unintended effects of androgen deprivation therapy and develop optimal strategies to prevent osteoporosis, obesity and obesity related disease. PMID- 15535445 TI - The impact of patient advocacy: the University of California-San Francisco experience. AB - PURPOSE: Although the principal goals of health care advocacy are to raise awareness of specific issues and make them national priorities, it is important that physicians should recognize that advocacy is a multidimensional phenomenon. In 1997 a group of scientists, administrators, clinicians and patients met to discuss individual goals, needs and areas of mutual interest in the areas of prostate cancer care and research. A formal advocacy group was established and advocates were integrated into the administration and planning process in specific areas of the clinical and research program. MATERIALS AND METHODS: In an effort to quantitate and communicate the impact of this group we assessed outcomes in the last 5 years in 4 areas matched to group interests and priorities, namely research, clinical trials, patient services and philanthropy. RESULTS: Improvements in all assessed metrics were noted (18% to 1,796%), most substantially in the areas of research grants (286%) and philanthropy (1,796%). The highlight of the collaborative effort was the awarding of the University of California-San Francisco Prostate Cancer SPORE, which included a highly rated, formal advocacy core. Advocates initiated at least 25 new initiatives to improve patient services in the areas of access to care, patient education, psychosocial support, outreach, etc. CONCLUSIONS: Since the initiation of a prostate cancer advocacy effort at our institution, substantial and quantitative improvements in several key metrics have been identified. Although an exact cause and effect cannot be stated with certainty, it seems likely. The successful collaboration was based on a multidimensional patient advocacy effort. PMID- 15535446 TI - On metaphor and melancholy. PMID- 15535447 TI - Daptomycin: overview of an innovative antibiotic. PMID- 15535448 TI - Damage control for blunt hepatic trauma: case presentation and historical review. AB - A 15-year old female with a Grade V liver injury from blunt trauma underwent therapeutic packing for uncontrolled hemorrhage. Pack removal in 72 hours following transfer to a major trauma center showed complete hemostasis. Six years later, CT scans revealed minimal liver sequella. Damage control surgery with initial therapeutic packing in blunt hepatic trauma is a valuable option for the community surgeon. PMID- 15535449 TI - The epidemiology of alcoholic liver disease. AB - This article describes the various forms of alcoholic liver disease (ALD), with particular emphasis on cirrhosis, the form of liver disease that often is most associated with alcohol abuse and about which the most information is available. Epidemiological research has evaluated the prevalence of ALD and the factors that often contribute to the disease. Although the most potent factor in ALD is the excessive consumption of alcoholic beverages, gender and ethnic differences also account for some important variations in rates of liver disease. Mortality rates from cirrhosis have declined in the United States and some other countries since the 1970s. A number of factors may have contributed to this decline, including increased participation in treatment for alcohol problems and Alcoholics Anonymous membership, decreases in alcohol consumption, and changes in the consumption of certain types of alcoholic beverages. PMID- 15535450 TI - Relationships between nutrition, alcohol use, and liver disease. AB - Many alcoholics are malnourished, either because they ingest too little of essential nutrients (e.g., carbohydrates, proteins, and vitamins) or because alcohol and its metabolism prevent the body from properly absorbing, digesting, and using those nutrients. As a result, alcoholics frequently experience deficiencies in proteins and vitamins, particularly vitamin A, which may contribute to liver disease and other serious alcohol-related disorders. Furthermore, alcohol breakdown in the liver, both by the enzyme alcohol dehydrogenase and by an enzyme system called the microsomal ethanol-oxidizing system (MEOS), generates toxic products such as acetaldehyde and highly reactive, and potentially damaging, oxygen-containing molecules. These products can interfere with the normal metabolism of other nutrients, particularly lipids, and contribute to liver cell damage. Nutritional approaches can help prevent or ameliorate alcoholic liver disease. For example, a complete balanced diet can compensate for general malnutrition. Administration of antioxidants (e.g., precursors of the endogenous antioxidant glutathione) can help the body eliminate reactive oxygen molecules and other reactive molecules generated from abnormal lipid breakdown. New agents currently are being studied as promising nutritional supplements for alcoholics with liver disease. PMID- 15535451 TI - Hepatitis C and alcohol. AB - Patients infected with the hepatitis C virus (HCV) who drink heavily are likely to suffer more severe liver injury, promoting disease progression to cirrhosis and increasing their risk for liver cancer. Some research, although not conclusive, suggests that even moderate drinking may spur liver damage in HCV infected patients. Research areas that have the greatest potential for developing more effective treatment options include HCV virology, immunology, animal models, and the mechanisms of liver injury. PMID- 15535452 TI - Hepatic encephalopathy. AB - Hepatic encephalopathy (HE) is a brain disorder caused by chronic liver failure, particularly in alcoholics with cirrhosis, which results in cognitive, psychiatric, and motor impairments. In these patients, the number of functional liver cells is reduced, and some blood is diverted around the liver before toxins are removed. As a result, toxins such as ammonia and manganese can accumulate in the blood and enter the brain, where they can damage nerve cells and supporting cells called astrocytes. Positron emission tomography analyses have determined that ammonia levels are elevated in the brains of HE patients; ammonia accumulation can alter the expression of various important brain genes. Magnetic resonance images show that manganese is deposited in a brain area called the globus pallidus; manganese deposits may be responsible for structural changes in the astrocytes that are characteristic of HE. Treatment of patients with HE involves measures to lower ammonia levels in the blood, medications to counteract ammonia's effects on brain cell function, devices to compensate for liver dysfunction, and liver transplantation. PMID- 15535453 TI - Diagnosis and treatment of alcoholic liver disease and its complications. AB - Alcoholic liver disease (ALD) is a serious and potentially fatal consequence of alcohol use. The diagnosis of ALD is based on drinking history, physical signs and symptoms, and laboratory tests. Treatment strategies for ALD include lifestyle changes to reduce alcohol consumption, cigarette smoking, and obesity; nutrition therapy; and pharmacological therapy. The diagnosis and management of the complications of ALD are important for alleviating the symptoms of the disease, improving quality of life, and decreasing mortality. PMID- 15535454 TI - Liver transplantation for alcoholic liver disease. AB - In many patients, long-term heavy drinking leads to chronic liver disease, liver failure, and even death. Orthotopic liver transplantation (OLT) is the only definitive treatment for end-stage liver disease, including alcoholic liver disease (ALD). Because of a shortage of donor organs, OLT for ALD patients remains controversial out of concerns that patients may resume drinking, thereby harming the transplanted organ. Therefore, transplant centers conduct careful screening procedures that assess patients' coexisting medical problems and psychosocial status to identify those patients who are medically most suited for the procedure and who are most likely to remain abstinent after OLT. Studies assessing the outcomes of ALD patients after OLT found that the survival rates of the transplanted organ and the patient were comparable to those of patients with nonalcoholic liver disease and that relapse rates among the ALD patients were low. Similarly, ALD patients and patients with other types of liver disease had comparable rates of compliance with complex medication regimens after OLT. Enhanced efforts to identify risk factors for relapse among OLT candidates with ALD and to target interventions specifically to those patients who are at high risk of relapse may further improve patient outcome and enhance the acceptance of OLT for alcoholic patients in the general population. PMID- 15535455 TI - Abo blood groups and completed reproductive performance of rural Haryanavi couples: analysing measures of selection intensities. AB - The possible differential effects of ABO blood group materno-paternal (fetal) incompatibility on completed reproductive performance were investigated on a sample of 100 couples (100 fathers and 100 mothers) from three villages in the Jind district of Haryana state, India. The average number of live births per mating couple was slightly higher for the incompatible matings (5.32) than the compatible ones (5.05). This advantage was offset by higher postnatal mortality in the former. Consequently, the average number of living children in the compatible matings (4.64) was higher than in the incompatible ones (4.18). With reference to individual ABO matings, the index of relative fertility (Irf) was the highest in A x AB followed by B x A type of incompatible matings. No decrease in live births in O x A and O x B incompatible matings was observed compared with their reciprocal compatible ones, i.e. A x O and B x O matings, as has been hypothesized in previous studies. The total pregnancy wastage was substantially higher in ABO-incompatible matings (24.59%) than compatible matings (8.45%). About 71% of the postnatal deaths took place within one year of the birth in the case of incompatible matings compared with 50% in the case of compatible matings. The study supports the hypothesis that selection is operative at the ABO locus as revealed by the measures of selection intensity. The loss of fitness in the present sample was associated with differential mortality. There were no differences in the proportions of average number of male live births in the compatible (0.55) and incompatible matings (0.58). However, in the individual mating types, there was some evidence of higher or lower proportions of male live births. PMID- 15535456 TI - Oral contraceptive non-compliance in rural Bangladesh. AB - This paper examines incorrect use of oral contraceptives (OCs) in rural Bangladesh by using data from an OC compliance survey. Of the 1031 current users of OCs interviewed, about 13% took their pills out of sequence, while 17% left incorrect intervals between pill packs. Forty per cent of the women reported missing one active pill during the 6 months prior to the survey, and 74% of them took correct action with the missed pill. Of the women who missed two active pills (16%), only 9% took correct action. Multivariate analyses revealed that women's education and their husbands' support helped protect against taking incorrect action with a missed pill. The fieldworker's contact was found to protect against leaving an incorrect interval between pill packs. Women who had membership of non-government organizations were less likely to interrupt their pill use, and more likely to take their pill out of sequence. The present study underscores the need for providing women with more support in their pill use, and advocates that service providers should be the focal point of efforts. Husbands' support is essential to improve the pill-taking behaviour of Bangladeshi women. PMID- 15535457 TI - Parental perceptions of costs and benefits of children as correlates of fertility in Kuwait. AB - Kuwait is a high fertility country where the average number of desired children still exceeds 5. However, fertility behaviour is beginning to show a noticeable change and the current TFR is about 4.2 children. In order to understand the decline in fertility, the impact of perceived benefits and costs of children on Kuwaiti women's desired and achieved fertility is analysed. Data from a nationally representative survey of Kuwaiti households held in 1999 are used. Bivariate analyses show that the mean desired number of children is significantly higher among those who rank higher on the perceived value of children, and lower among those who are concerned about the cost of raising children. Achieved fertility (children ever born) shows a similar pattern. In the multivariate analysis, however, large family values emerge as the only significant predictor of (higher) desired as well as achieved fertility, net of the respondent's demographic and socioeconomic characteristics. The sociocultural, economic and political contexts that shape the mother's perceptions of the benefits and costs of children are analysed and it is concluded that the need for children as social and national capital is currently the most important driving force behind fertility desires and behaviour. PMID- 15535458 TI - Female sterilization in Latin America: cross-national perspectives. AB - Fertility levels have dropped substantially in Latin America in recent decades, fuelled by increased contraceptive use and notably a method mix skewed towards female sterilization. This study examined choice of female sterilization in four Latin American countries: Brazil, Colombia, the Dominican Republic and Peru. Data were drawn from national Demographic and Health Surveys conducted in 1995--1996. Discrete-time hazard models were used on the five-year calendar modules of women's reproductive histories to consider the effects of a number of sociodemographic and contextual determinants as they pertained to status at the moment of the event. The results revealed that the likelihood of a woman's having undergone contraceptive sterilization was increasing over time in Brazil and Peru, suggesting that the potential for future growth of this method remains strong. A consistent pattern of increased probability of sterilization with higher education was seen across all countries, seemingly dispelling certain controversial claims that the procedure may have been disproportionately performed on the poor. At the same time, the multilevel analytical approach pointed to significant cluster-level random effects, suggesting that there were additional unmeasured contextual influences on women's propensity to choose sterilization. PMID- 15535459 TI - The role of social factors and weight status in ideal body-shape preferences as perceived by Arab women. AB - This study investigated the social factors associated with body-shape preferences for females and males as perceived by Arab women living in Qatar, and correlated the current weight status of women studied with these preferences. The subjects were 535 non-pregnant Arab women aged 20-67 years, who attended heath centres in Doha City, the capital of the State of Qatar. Illustrations of male and female body shapes ranging from very thin to very obese using the 9-figure Silhouettes scale were shown to women, and they were asked to select their preferred figure. Body mass index (BMI) was used to determine the weight status of women studied. Age, educational level and employment status were found to be significantly associated with ideal body-shape preference for both males and females, whereas marital status and current weight status had no significant association. In general, the Arab women studied selected a more mid-range of body fatness for males than for females. It is concluded that attention should be given to sociocultural factors, such as body-shape preferences, in any programmes to promote ideal body weight for the public. PMID- 15535460 TI - Recent trends and components of change in fertility in Nepal. AB - The objectives of this article are, first, to provide improved estimates of recent fertility levels and trends in Nepal and, second, to analyse the components of fertility change. The analysis is based on data from Nepal's 1996 and 2001 Demographic and Health Surveys. Total fertility rates (TFR) are derived by the own-children method. They incorporate additional adjustments to compensate for displacement of births, and they are compared with estimates derived by the birth-history method. Fertility is estimated not only for the whole country but also by urban/rural residence and by woman's education. The own-children estimates for the whole country indicate that the TFR declined from 4.96 to 4.69 births per woman between the 3-year period preceding the 1996 survey and the 3 year period preceding the 2001 survey. About three-quarters of the decline stems from reductions in age-specific marital fertility rates and about one-quarter from changes in age-specific proportions currently married. Further decomposition of the decline in marital fertility, as measured by births per currently married woman during the 5-year period before each survey, indicates that almost half of the decline in marital fertility is accounted for by changes in population composition by ecological region, development region, urban/rural residence, education, age at first cohabitation with husband, time elapsed since first cohabitation, number of living children at the start of the 5-year period and media exposure. With these variables controlled, another one-third of the decline is accounted for by increase in the proportion sterilized at the start of the 5 year period before each survey. PMID- 15535461 TI - The growth of IQ among Estonian schoolchildren from ages 7 to 19. AB - The Standard Progressive Matrices test was standardized in Estonia on a representative sample of 4874 schoolchildren aged from 7 to 19 years. When the IQ of Estonian children was expressed in relation to British and Icelandic norms, both demonstrated a similar sigmoid relationship. The youngest Estonian group scored higher than the British and Icelandic norms: after first grade, the score fell below 100 and remained lower until age 12, and after that age it increased above the mean level of these two comparison countries. The difference between the junior school children and the secondary school children may be due to schooling, sampling error or different trajectories of intellectual maturation in different populations. Systematic differences in the growth pattern suggest that the development of intellectual capacities proceeds at different rates and the maturation process can take longer in some populations than in others. PMID- 15535462 TI - Diet effects on female reproduction in high growth (hg/hg) mice that are deficient in the Socs-2 gene. AB - The detrimental effect of larger body size on reproductive performance has been well documented in mouse models of overgrowth, such as Growth Hormone (GH) transgenics. This study describes the reproductive performance of the High Growth (HG) mouse model of overgrowth. The HG mouse model exhibits overgrowth due to a partially recessive autosomal mutation that increases growth rate and mature body size. The HG phenotype results from the lack of expression of Socs-2, which negatively regulates GH signaling. C57BL/6J (C57) and congenic C57BL/6J-hg/hg (HG) female mice were fed four diets differing in protein and energy content, starting at 8 weeks of age. A complete reproductive cycle from mating to weaning was evaluated. HG mice were 40% larger than C57 and had a higher feed intake throughout the experiment. Significant genotype x diet interactions were detected for growth, body composition and reproductive traits. HG females showed poor reproductive performance compared to controls as demonstrated by their lower fertility during mating, which was not overcome by changes in the diet. No differences were detected in litter size, but HG animals exhibited a longer gestation length and heavier pup weaning weights compared to controls. Reproductive impairment in HG seems to be the consequence of the lack of Socs-2 independent of the effects of increased body size on reproduction. PMID- 15535463 TI - Effect of anaerobic fungi on in vitro feed digestion by mixed rumen microflora of buffalo. AB - Five strains of anaerobic fungi isolated from the faeces of wild (hog deer, Cervus porcinus; blackbuck, Antelope cervicapra; spotted deer, Axis axis; nilgai, Baselophus tragocamelus) and rumen liquor of domestic (sheep, Ovies aries) ruminants showing high fibrolytic enzyme producing ability were added to mixed rumen microflora of buffalo to study their effect on the digestibility of lignocellulosic feed (wheat straw and wheat bran in the ratio of 80:20), enzyme production and fermentation end products in in vitro conditions. Among the 5 isolates studied, FNG5 (isolated from nilgai) showed the highest stimulating effect on apparent digestibility (35.31 +/- 1.61% vs. 28.61 +/- 1.55%; P < 0.05), true digestibility (43.64 +/- 1.73% vs. 35.37 +/- 1.65%; P < 0.01), neutral detergent fiber digestibility (29.30 +/- 2.58% vs. 18.47 +/- 2.12; P < 0.01) of feed 24 h after inoculation compared to the control group. The production of carboxymethyl cellulase, xylanase, acetyl esterase and beta-glucosidase was significantly (P < 0.05) higher in the FNG5 inoculated incubation medium. There was no improvement in the digestibility and enzyme production on the addition of the other 4 isolates. Total volatile fatty acid levels as well as the concentration of acetate, propionate, isobutyrate and valerate were significantly higher in the FNG5 added group as compared to the control group. The fungal isolate FNG5 from nilgai, a wild ruminant, was found to be superior to the other isolates tested and appears to have a potential to be used as a feed additive for improving fiber degradation in domestic ruminants. PMID- 15535464 TI - Effect of feeding program during rearing and age at first insemination on performances during subsequent reproduction in young rabbit does. AB - An experiment was performed to study the effect of the feeding program and age at first mating on body growth, feed intake, reproductive performance, and culling of rabbit does over three parities, using 155 does of a strain of New Zealand white rabbits. Three treatments were applied. Ad libitum feeding until first insemination at 14.5 wk (AL-14.5) or 17.5 wk of age (AL-17.5), and restrictive feeding from five wk of age until first insemination at 17.5 wk of age (R-17.5). At first insemination, the BW of AL-14.5 and R-17.5 was similar (3 907 vs. 3 791 +/- 46 g, respectively), whereas AL-17.5 does were heavier (4 390 +/- 46 g, P < 0.001). During reproduction, performance of AL-17.5 was not improved compared to AL-14.5 and R-17.5 does. Al-17.5 does showed a lower feed intake during the first gestation (-25%) and first parity (-10%) than R- 17.5, resulting in weight loss ( 6%) during the first gestation and decreased litter weights (-19%) and litter growth (-14%) in the first parity. Extended first mating by three wk (17.5 vs. 14.5 wk) but similar BW at first mating did not affect feed intake and BW development during the first three parities. However, the number of live born kits and weight at first kindling, and litter growth in the first parity were improved in R-17.5 (+23%, +18%, and +14%, respectively). Reproductive performance can be improved by restricted feeding during rearing and extended first insemination to 17.5 wk of age. However, the culling rate was not affected by the rearing strategy. PMID- 15535465 TI - Ovulation rate, litter size and prenatal losses in hair sheep of the French West Indies. AB - Ovulation rate (OR) and litter size (LS) were recorded in local hair sheep of Martinique and in local x Lacaune-viande crossbreeds, in order to assess prenatal losses. Local hair sheep LS and OR were 1.91 and 2.41, respectively. Prenatal losses increased with OR, from 0.22 (OR = 2) up to 1.50 (OR > 3). The ewes (69.7%) with LS = 1 had actually lost at least one ovum, against 25.9% of the ewes with LS > 1. Ewes grazing Cynodon nlemfuensis (Stargrass) had lower LS than those grazing Digitaria decumbens (Pangola), in relation with a lower nutritional level and, maybe, more parasitism effects. No seasonal or age at lambing effect was shown. The OR and LS were dramatically decreased on local x Lacaune-viande crossbred ewes (minus 1.32 and 0.67 unit, respectively, P < 0.05). This suggests a possible negative effect of the tropical environment on the crossbreeds' reproductive function. PMID- 15535466 TI - Regulation of seasonality in the migratory male blackheaded bunting (Emberiza melanocephala). AB - The present study was carried out on a Palearctic-Indian migratory species, the blackheaded bunting (Emberiza melanocephala), to understand the importance of photoperiodism and circannual rhythms in determining seasonality in changes in body mass and testis size in birds. An initial experiment determined the effects of duration and intensity of light on photoperiodic induction. The birds were exposed to different photoperiods (hours of light:hours of darkness; 11.5L:12.5D, 12L:12D, 12.5L:11.5D and 13L:11D) at the same (approximately 450 lux) light intensity, and to 13L:11D at different light intensities (50-, 100-, 400-, 800- and 1000-lux). The induction and subsequent regression of photoperiodic responses were dependent upon duration and intensity of the light period until these reached threshold. A second experiment investigated if an endogenous seasonal rhythm underlies photoperiodism in buntings. Birds maintained since February on a 8L: 16D photoperiod (a non-inductive short day length invariably used to ensure photosensitivity in photoperiodic species) were subjected periodically to 16L:8D (a long day length), one group every month from mid-March to mid-August. The magnitude of long day response in body mass and testes decreased as the duration of the short days progressed, but testicular response was restored in birds that were exposed to long days in July and August. The birds exposed simultaneously to short, long, and natural day lengths for 32 weeks underwent an induction regression cycle under long days and natural day lengths, but not under short days in which a decrease in body mass occurred after about 20 weeks. The last experiment examined the importance of latitudinal migration on photoperiodism, by comparing the response to long days of three groups which included birds from populations those were held in the outdoor aviary for 1 or 2 years at 27 degrees N and those immediately arrived from their breeding grounds (approximately 40 degrees N). There was no difference in the photoperiodic induction among the three groups, indicating that neither experience to changing photoperiods during a migratory journey, nor to long photoperiods at breeding grounds, were critical for a subsequent response (initiation-termination-reinitiation) cycle. Taken together, these findings suggest that (1) the blackheaded bunting has its own endogenous timing program, which is regulated by the photoperiod, and (2) the photoperiodic programs of bunting are flexible enough to accommodate variations in the amplitude of environmental cycles. Thus, it appears that photoperiodism has evolved independently of the evolution of migration in this species. PMID- 15535467 TI - Short-term effects of spermine ingestion on the small intestine: a comparison of suckling and weaned rats. AB - We have previously shown that spermine, shortly after its ingestion, can induce the alteration of the morphology of the small intestine of suckling rats. It was proposed that this alteration is due to polyamine accumulation inside the epithelial cells. This could also be related to the fact that the intestine of the suckling rat is in an immature state. To shed light on this issue, disaccharidase and alkaline phosphatase activity assays, protein, DNA and RNA content measurements and polyamine concentration analysis were performed on the small intestine of suckling and weaned Wistar rats treated with spermine. Spermine did not induce the same intestinal alterations in weaned rats compared to suckling animals. Indeed, in sucklings, spermine administration induced a decrease of the protein, DNA, putrescine and spermidine intestinal content, suggesting a cell loss. The cell loss impaired the activity of intestinal enzymes: lactase, maltase and alkaline phosphatase. In weaned rats, the same treatment did not alter these parameters. Exogenous spermine by itself is not sufficient to induce the alterations described here and previously. The maturity degree of the small intestine could be the basis of this process. PMID- 15535468 TI - Photosensitivity in body mass and testicular activity of brahminy myna, Sturnus pagodarum. AB - This study analyzed photoperiodic sensitivity of brahminy myna (Sturnus pagodarum), which is a seasonally breeding bird species. During regression phase of the reproductive cycle (in early September), groups of myna were exposed to artificial photoperiods that were either close to or longer than those brahminy myna experiences at this time in wild at 29 degrees N. Following a 14-week exposure to such photoperiods (hours of light: hours of darkness; 13L:11D, 12L:12D, 11.5L:12.5D and 11L:13D), the birds were subjected to a longer day length (16L:8D) for another 9 weeks to test whether pre-treatment with varying photoperiods had an effect on subsequent long day photostimulation. There was a progressive increase in body mass under different pre-treatment photoperiods, with a faster increase in 11L:13D and 11.5L:12.5D than in 12L:12D and 13L:11D. When subjected to 16L: 8D, however, all groups showed decline in body mass. By contrast, the testes were not stimulated under 11L:13D and 11.5L:12.5D, and only one individual of the groups under 12L:12D and 13L: 11D showed a small testis recrudescence. All birds except one individual of the 13L:11D group, however, showed testis recrudescence when subjected to 16L:8D. These results suggest that body mass and testes have a different profile of photoperiodic response and appear to have two different threshold photoperiods for dissipating the post reproductive refractoriness. PMID- 15535469 TI - Dose-effect of dietary oleic acid: oleic acid is conditionally essential for some organs. AB - The minimum dietary intake of oleic acid that is indispensable to maintain a normal content of this fatty acid in several tissues (heart, muscle, kidney and testis) was determined in the rat. For this purpose, a dose-effect study was conducted using an experimental protocol with 7 groups of rats who received a diet in which the oleic acid level varied from 0 to 6000 mg per 100 g diet, but the other ingredients were identical (in particular the essential fatty acids, linoleic and alpha-linolenic acid). Female rats were fed the diets from two weeks before mating, and their pups were killed aged either 21 or 60 days. When the level of oleic acid in the diet was increased, the main modifications observed in 21-day-old deficient pups were as follows: (i) for 18:1n-9, in the liver, muscle, heart, kidney, and testis, a plateau was reached at about 4 g oleic acid per 100 g diet. Below this level, the higher the dose the greater the response; (ii) for 16:1n-7, the concentration decreased in the liver, muscle, heart, kidney and testis; (iii) the concentration of 18:1n-7 decreased in the kidney, muscle, and testis; (iv) some minor modifications were noted for the other fatty acids. In mother's milk at 14 days of lactation, when dietary oleic acid increased, the levels of 18:1(n-9) also increased; the increase was regular and did not reach a plateau. In 60-day-old rats, the results were generally similar to those in 21 day-old rats, but with some differences, in particular a slight decrease in oleic acid concentration in the liver and kidney at the highest dietary oleic acid level. PMID- 15535470 TI - Hormones, IgG and lactose changes around parturition in plasma, and colostrum or saliva of multiparous sows. AB - Blood, colostrum and saliva samples were serially taken from 6 multiparous sows from day 109 of gestation until day 3 postpartum. Plasma was assayed for oestradiol-17beta (E2), progesterone (P4), prolactin (PRL), cortisol, immunoglobulin G (IgG) and lactose. Colostrum was assayed for E2, P4, IgG and lactose. Lactoserum, obtained after ultra centrifugation of colostrum, was assayed for PRL. Saliva was assayed for cortisol. Time-related variations in hormone, IgG and lactose concentrations measured in plasma were parallel to those measured in colostrum, lactoserum or saliva. However, the concentrations were higher in colostrum or lactoserum and lower in saliva than in plasma. Ratios of concentrations of cortisol in saliva and PRL in lactoserum over those in plasma did not vary with time and averaged 0.2 and 1.6, respectively. Conversely, the ratios of concentrations of E2 and P4 in colostrum over those in plasma varied with time (P < 0.05) but were quite constant before the end of parturition, averaging 2.7 and 3.6, respectively. The ratios of concentrations of IgG and lactose in colostrum over those in plasma also varied with time (P < 0.05). The concentrations of hormones in plasma on the one hand and in colostrum, lactoserum or saliva on the other hand were significantly correlated but correlations varied with time (PRL across periods: r = 0.31; cortisol across periods: r = 0.60; E2 during parturition: r = 0.83; P4 before parturition: r = 0.82; P4 during parturition: r = 0.67). The present results indicate that around parturition, assays of hormones in colostrum or saliva can be used to study the hormonal status of sows. Furthermore, variations in colostrum and plasma concentrations of IgG and lactose are good indicators of the transition from colostrum to milk synthesis. PMID- 15535471 TI - A commentary on the history of social psychiatry and psychotherapy in twentieth century Germany, Holland and Great Britain. PMID- 15535472 TI - Between institutional psychiatry and mental health care: social psychiatry in The Netherlands, 1916-2000. PMID- 15535473 TI - Psychotherapy in The Netherlands after the Second World War. PMID- 15535474 TI - Social psychiatry in Germany in the twentieth century: ideas and models. PMID- 15535475 TI - Psychotherapy between medicine, psychoanalysis, and politics: concepts, practices, and institutions in Germany, c. 1945-1992. PMID- 15535476 TI - War and the practice of psychotherapy: the UK experience 1939-1960. PMID- 15535477 TI - The validity of board exams. PMID- 15535478 TI - Special care dentistry: it's time to get in the ring. PMID- 15535479 TI - Special needs patients--almost 107,000 children with disabilities in Pennsylvania. AB - The 2000 Census data indicating the numbers of children with disabilities in Pennsylvania is presented and broken down by city, metropolitan area and congressional districts. There are numerous difficulties associated with the delivery of dental care to these youngsters with special needs. Nevertheless, if all dentists were willing to help, each practitioner would need to care for only 14 of these youngsters. PMID- 15535480 TI - Service on the road less traveled. PMID- 15535481 TI - New overtime rules: payroll/human resources (HR) priority. PMID- 15535482 TI - Local anaesthetics--the origins of lignocaine. PMID- 15535483 TI - The role of microvascular thrombosis in sepsis. AB - The acute inflammatory response to sepsis gives rise to significant morbidity and mortality. The mechanisms underlying this form of tissue injury are poorly understood. This review examines the evidence that tissue ischaemia due, to generalized microvascular thrombosis may play an important role. Microvascular thrombosis is probably an adaptive response that prevents bacteria in the tissues reaching the systemic circulation via the capillaries. In time, a definitive response by leucocytes removes the bacteria and repairs the damaged tissues. There is however evidence that if microvascular thrombosis becomes generalized, then extensive tissue ischaemia may precipitate organ failure and death. Post mortem studies of patients with sepsis demonstrate microvascular thrombi in many organs including the kidney, liver, lung, gut, adrenals and brain, and the degree of organ injury is related to the quantity of thrombi. Furthermore studies in human and animal models of sepsis demonstrate therapies that inhibit coagulation or promote fibrinolysis reduce organ failure and mortality. In view of the personal and economic burdens that tissue injury associated with the acute inflammatory response places on the community, further studies to establish the role of microvascular thrombosis are clearly required. Such studies may lead to new therapies to limit or prevent this form of injury. PMID- 15535484 TI - Delayed epidural catheter removal: the impact of postoperative coagulopathy. AB - A retrospective analysis of 413 patients who received postoperative epidural analgesia under a standardized protocol found that 84 (20%) had a duration of epidural catheterization of greater than four days. The most common reasons were significant pain (n=64, 15%) and coagulopathy (n=26, 6%). Risk factor analysis for coagulopathy showed an odds ratio of 10.1 (95% confidence interval 4.2-24.5) for prolonged epidural catheterization among patients undergoing hepatectomy. Magnetic resonance imaging, performed in four patients with clinical signs suggestive of epidural haematoma, was negative for a space-occupying lesion in all cases. Eleven patients developed fever and clinical signs suggestive of epidural catheter-related infection, necessitating early catheter removal. Sixteen patients had persistent lower limb weakness at 24 hours after catheter removal. The signs soon resolved in all except two, one of whom had neuropathy related to intraoperative positioning and the other preoperative weakness. Accidental epidural catheter dislodgement occurred in 29 patients (7%) and is potentially hazardous if coagulopathy is unresolved. The risk-benefit ratio and factors complicating catheter removal, especially coagulopathy, should be considered when deciding whether to use epidural techniques. PMID- 15535485 TI - Monitoring of acid-base and regulating variables during abdominal lavage. AB - This prospective clinical study was performed to determine acid-base regulating variables during abdominal lavage treatment for patients with severe peritonitis or after abdominal surgery. Arterial blood was sampled from twelve patients with secondary peritonitis and nine patients after abdominal surgery without peritonitis at three time points: immediately before, immediately after and 15 minutes after abdominal lavage with normal saline solution. The total amount of irrigant fluid, the strong ion difference [(Na+ + K+)-(Cl- + lactate-)], and total protein concentrations were determined and standard bicarbonate, standard base excess were calculated from pH and PaCO2. Peritonitis patients developed a moderate alkalaemia (pH 7.440-7.485). The alkalaemia was unmasked after optimization of mild hypoventilation, but was supported by a decrease in protein concentration of about 3.4 mEql/l in the first 15 minutes after the lavage. There was no marked increase in chloride concentration in either the peritonitis or the control group. The data indirectly exclude major fluid absorption during abdominal lavage with 3000 to 6000 ml normal saline, given that we found no clinically relevant electrolyte and acid-base changes that might be expected after rapid fluid absorption. The factors of major influence in acid-base regulation were ventilation and protein loss in the course of abdominal lavage. Monitoring of the Stewart variables is an easily applicable method of monitoring acid-base regulating variables in the perioperative course of patients undergoing abdominal lavage therapy. PMID- 15535486 TI - The effects on arterial haemoglobin oxygen saturation and on shunt of increasing cardiac output with dopamine or dobutamine during one-lung ventilation. AB - Theoretically, if the cardiac output were increased in the presence of a given intrapulmonary shunt, the arterial saturation should improve as the venous oxygen extraction per ml of blood decreases if the total oxygen consumption remains constant. Previous work demonstrated that this was not achieved with adrenaline or isoprenaline as increased shunting negated any benefit from improved cardiac output and mixed venous oxygen content. However pharmacological stimulation of cardiac output and venous oxygen without any increase in shunt should achieve the goal of improved arterial oxygenation. To test this hypothesis, seven pigs were subjected to one-lung ventilation and infused on separate occasions, with dopamine and with dobutamine in random order to increase the cardiac output. The mixed venous oxygen content, shunt fraction, oxygen consumption and arterial oxygen saturation were measured. With both dopamine and dobutamine there was a consistent rise in venous oxygen content. However, with dopamine, the mean shunt rose from 28% to 42% and with dobutamine, the mean shunt rose from 45% to 59% (both changes P<0.01). With dopamine, the mean arterial oxygen saturation fell by 4.7%, and with dobutamine by 2.9%, but neither fall was statistically significant. It is concluded that any benefit to arterial saturation which might occur from a dopamine- or dobutamine-induced increase in mixed venous oxygen content during one-lung ventilation is offset by increased shunting. During one lung anaesthesia, there would appear to be no benefit to arterial saturation in increasing cardiac output with an infusion of either dopamine or dobutamine. PMID- 15535487 TI - Early and late reversal of rocuronium with pyridostigmine during sevoflurane anaesthesia in children. AB - This study investigated the effect of pyridostigmine administered at different levels of recovery of neuromuscular function after rocuronium during sevoflurane anaesthesia in children. Fifty-one patients aged 3 to 10 years, ASA physical status 1 or 2 were randomized to 4 groups: a spontaneous recovery group; or, reversal with pyridostigmine 0.25 mg/kg with glycopyrrolate 0.01 mg/kg at one of three times: 5 minutes after rocuronium administration; at 1% twitch height (T1) recovery; or at a 25% twitch height (T25) recovery. Anaesthesia was induced with thiopentone (5-7 mg/kg) and maintained with 2-3% sevoflurane and 50% nitrous oxide. Atropine (0.015 mg/kg) and, after calibrating the TOF-Watch, rocuronium (0.6 mg/kg) were then administered. Maximal block occurred 1.1+/-0.5 min (mean, SD) after rocuronium administration. In the spontaneous recovery group, the clinical duration (recovery to T25) was 40.1+/-8.8 min and the recovery index (time between T25 and T75) 19.9+/-9.8 min. Recovery to TOF >0.9 from the time of rocuronium administration was reduced by approximately 30% in the pyridostigmine groups compared to the spontaneous recovery group. There was no significant difference among the three pyridostigmine groups. When pyridostigmine was given at T1 or T25, the time from pyridostigmine administration to TOF >0.9 was shorter than for the group receiving pyridostigmine 5 minutes after rocuronium. PMID- 15535488 TI - Effect of metoclopramide on pain on injection of propofol. AB - We undertook a randomized, double-blind, placebo-controlled study to examine the efficacy of metoclopramide at three different doses (2.5 mg, 5 mg, 10 mg) for reducing pain on injection of propofol in 100 patients scheduled for elective surgery. Patients received intravenously the study drug, with venous occlusion for one minute, followed by propofol 2 mg/kg into a dorsal hand vein. The incidence of pain was significantly less in patients receiving metoclopramide 5 mg (32%) or 10 mg (28%) than in patients receiving placebo (80%) (P<0.01). No difference between metoclopramide 2.5 mg and the placebo groups was found. We conclude that pretreatment of a dorsal hand vein with metoclopramide in a dose of 5 or 10 mg, with venous occlusion for one minute, effectively decreases the incidence of pain caused by propofol injection. PMID- 15535489 TI - Pain during injection of propofol: the effect of prior administration of ephedrine. AB - Propofol causes pain on intravenous injection in 28 to 90% of patients. A number of techniques have been tried to minimize propofol-induced pain, with variable results. In a randomized, double-blind, placebo-controlled trial, we compared the efficacy of ephedrine 30 microg/kg pretreatment to lignocaine 40 mg for prevention of propofol-induced pain. Ninety-three adult patients, ASA 1 and 2, undergoing elective laparoscopic cholecystectomy were randomly assigned to three groups of 31 each. Group 1 received normal saline, group 2 received lignocaine 2% (40 mg) and group 3 received 30 microg/kg ephedrine. All pretreatment drugs were made up to 2 ml. Pain at the time of propofol injection was assessed on a four point scale: 0=no pain, 1 =mild pain, 2=moderate pain, and 3=severe pain. Twenty seven patients (87%) of ephedrine pretreatment patients had pain during intravenous injection of propofol as compared to 24 (77%) in the normal saline group. In the lignocaine group, propofol-induced pain was observed in only 13 (42%) when compared with other study groups (P<0.05). Pretreatment with ephedrine 30 microg/kg did not attenuate pain associated with intravenous injection of propofol, nor did it improve haemodynamic stability during induction. However, pretreatment with 2% lignocaine (40 mg) was effective in attenuating propofol associated pain. PMID- 15535490 TI - Adjustment of anaesthesia depth using bispectral index prolongs seizure duration in electroconvulsive therapy. AB - Electroconvulsive therapy (ECT) under propofol anaesthesia induces relatively shorter seizures compared to barbiturate anaesthesia. Since significant correlation between seizure duration and bispectral index (BIS) value immediately before electrical stimulus has been reported among patients, adjustment of anaesthesia depth as determined by BIS may be effective in obtaining a longer seizure length. In the present study, we examined this hypothesis in those patients whose muscular seizure duration was less than 40s. ECT was prescribed to 20 patients suffering from endogenous depression. General anaesthesia was induced with propofol 1 mg/kg. Succinylcholine chloride 1 mg/kg was then given. The efficacy of electrical stimulation was determined using a tourniquet technique, electromyogram, and electroencephalography. When a patient had a seizure less than 40s in their second ECT treatment, the subsequent treatment was modified such that the electrical stimulus was given after waiting for a higher BIS value (+10-20). Intensity of electrical stimulus and anaesthesia conditions were identical in the two treatments. All 20 patients had longer seizures as determined by the electromyogram and/or electroencephalography when the stimulus was delivered at the higher BIS value. Seizure duration measured by muscle movement was 31+/-5 s when the stimulus was delivered without waiting and 46+/-10 s when delivered after waiting. There was a significant difference in seizure duration between the two treatments (P<0.01). Waiting for a recovery in BIS value before electrical stimulation can prolong seizure duration. PMID- 15535491 TI - Early experience with magnesium administration in Irukandji syndrome. AB - The administration of magnesium sulphate is a proposed novel therapy for Irukandji syndrome'. In this non-randomized, unblinded case series, data from ten patients who received magnesium salts are reviewed. Magnesium sulphate boluses of 10 to 20 mmol, in the six patients for which there was adequate data, reduced pain scores immediately after administration from 8.7+/-1.5 to 2.8+/-2.8 (Wilcoxon rank-sum test, P=0.03). In ten patients blood pressure decreased with a mean difference of -18 mmHg in mean arterial pressure. Magnesium requirements in individual patients varied markedly. Pain on injection occurred in four patients, three of whom had received peripherally administered magnesium chloride, and one patient reported transient ptosis after administration of magnesium sulphate 166 mmol over 18 hours in the setting of severe Irukandji syndrome. Magnesium sulphate administration appears to attenuate pain and hypertension in Irukandji syndrome and warrants further evaluation in this setting. PMID- 15535492 TI - Nasopharyngeal oxygen in adult intensive care--lower flows and increased comfort. AB - Nasopharyngeal oxygen therapy, the delivery of supplementary oxygen into the nasopharynx via a fine catheter placed through the nose, is a simple technique used in postoperative anaesthetic care units and paediatric intensive care, but never described in the setting of adult intensive care. In a prospective crossover design, we compared nasopharyngeal oxygen therapy with semi-rigid plastic mask (Hudson Mask) in 50 unintubated adult patients receiving supplemental oxygen. We measured oxygen flow rate to achieve cutaneous saturations 93 to 96%, and patient comfort by visual analogue score. Nasopharyngeal oxygen therapy consumed significantly less oxygen than mask administration (3.0+/-0.9 vs 6.7+/-2.1 l/min, P<0.001) and was associated with significantly higher comfort than the mask (7.5+/-1.6 cm vs 5.2+/-1.8, P<0.001). PMID- 15535493 TI - The use of end-tidal carbon dioxide monitoring to confirm endotracheal tube placement in adult and paediatric intensive care units in Australia and New Zealand. AB - The use of end-tidal carbon dioxide monitoring to assist in confirming endotracheal tube placement is currently not mandatory in intensive care units (ICUs) in Australia and New Zealand. Early detection of failed tracheal intubation is vital to optimize management and to prevent complications. Questionnaires were sent to the lead clinician/head of department of all 66 intensive care units approved for training purposes by the Joint Faculty of Intensive Care Medicine in Australia and New Zealand. The methods used to confirm correct endotracheal tube placement, the availability of end-tidal carbon dioxide monitoring and its role in confirming endotracheal tube placement in the intensive care unit were explored. Completed questionnaires were received from 61 of the 66 centres (92.4%). Wide variation in the method of confirmation of endotracheal tube position was demonstrated, with 23 (37.7%) of units using sub optimal methods. Sixty (98.3%) of units had end-tidal carbon dioxide monitoring available. Thirty-eight (62%) units shared monitors between several beds; and 22 (36%) had one monitor per bed. End-tidal carbon dioxide monitoring was used routinely to confirm endotracheal tube placement in 42 (68.8%) units. Fifty-two respondents (83.3%) felt that end-tidal carbon dioxide monitoring was superior to other methods for confirming endotracheal tube placement in critically ill patients. Thirty-eight respondents (62.3%) thought that end-tidal carbon dioxide monitoring should be mandatory to confirm tracheal intubation in the intensive care unit. If it were available, 42 respondents (68.8%) would use end-tidal carbon dioxide monitoring for confirmation of every intubation. Mandatory end tidal carbon dioxide confirmation of endotracheal tube placement was policy in 33 (54.1%) of the intensive care units. PMID- 15535494 TI - Suggestion, hypnosis and hypnotherapy: a survey of use, knowledge and attitudes of anaesthetists. AB - Clinical hypnosis is a skill of using words and gestures (frequently called suggestions) in particular ways to achieve specific outcomes. It is being increasingly recognised as a useful intervention for managing a range of symptoms, especially pain and anxiety. We surveyed all 317 South Australian Fellows and trainees registered with ANZCA to determine their use, knowledge of, and attitudes towards positive suggestion, hypnosis and hypnotherapy in their anaesthesia practice. The response rate was 218 anaesthetists (69%). The majority of respondents (63%) rated their level of knowledge on this topic as below average. Forty-eight per cent of respondents indicated that there was a role for hypnotherapy in clinical anaesthesia, particularly in areas seen as traditional targets for the modality, i.e. pain and anxiety states. Nearly half of the anaesthetists supported the use of hypnotherapy and positive suggestions within clinical anaesthesia. Those respondents who had experience of clinical hypnotherapy were more likely to support hypnosis teaching at undergraduate or postgraduate level when compared with those with no experience. PMID- 15535495 TI - Gum elastic bougie-guided insertion of the ProSeal Laryngeal Mask Airway. AB - We tested the hypothesis that gum elastic-bougie-guided insertion of the ProSeal Laryngeal Mask Airway is more frequently successful than introducer tool guided insertion after failed digital insertion. One hundred anaesthetized patients (ASA 1-2, aged 18 to 80 years) were randomized for the second insertion attempt using either the gum elastic bougie-guided or introducer tool techniques. The bougie guided technique involved priming the drain tube with the bougie, placing the bougie in the oesophagus using laryngoscope guidance, digital insertion along the palato-pharyngeal curve, and bougie removal. The introducer tool technique involved attaching the introducer tool, single-handed rotation along the palatopharyngeal curve, and introducer tool removal. Failed insertion was classified as (i) failed passage into the pharynx, (ii) malposition, or (iii) ineffective ventilation. Any blood staining was documented. Insertion was more frequently successful (50/50 vs 15/50, P=0.0002) and faster (35+/-17 s vs 54+/-45 s, mean+/-SD, P=0.006) with the bougie-guided technique. All failed insertions with the introducer tool technique were successful with the bougie-guided technique. The aetiology of failed insertion was similar for the digital and introducer tool techniques in 94% (33/35) of patients. There was no blood staining on the bougie, laryngoscope or introducer tool at removal, but blood staining was more common on the ProSeal Laryngeal Mask Airway with the introducer tool technique (9/50 vs 2/50, P=0.03). We conclude that the gum elastic bougie guided insertion has a higher success rate and causes less trauma than the insertion tool insertion technique after failed digital insertion of the ProSeal Laryngeal Mask Airway. PMID- 15535496 TI - The single-connector technique for initial placement of double-lumen tubes. AB - Due to the presence of major lung or extra-pulmonary pathology, which may be unilateral or bilateral, the initial placement of a double-lumen tube is not always straightforward. Although fibreoptic bronchoscopy is often used to confirm "correct" placement, a "blind" technique is frequently used for the initial insertion. The currently widely taught blind technique involves tracheal cuff inflation and ventilation of both lungs as a first manoeuvre, with a subsequent assessment of single-lung ventilation by clamping off in turn, the two limbs of the double-lumen tube double-connector: An alternative approach involves the bronchial cuff being inflated first, and then using a single-connector to transfer ventilation from one lung to the other. In this paper this technique is described and compared to the more traditional method. On a purely "number of steps" basis, the single-connector approach has several advantages. Furthermore, use of a technique that involves bronchial cuff inflation and single-lung ventilation as a first manoeuvre may reduce the risk of a temporarily malplaced double-lumen tube creating a potentially harmful ball-valve effect in a partially obstructed lobe or lung. PMID- 15535497 TI - An audit of the Single Use Portex Laryngeal Mask. AB - We performed an audit of the insertion of the Single Use Portex Laryngeal Mask in 400 patients. Insertion was successful at first attempt in 335 out of 400 patients (83.8%). However in 15 patients (3.8%), the Portex laryngeal mask could not be placed despite repeated attempts. In 12 of these 15 patients (80%), a standard Laryngeal Mask Airway (LMA) was successfully placed. After the completion of the audit, 22 out of 29 anaesthetists (75.9%), who had inserted > or = 5 Portex laryngeal masks, considered it inferior to the standard LMA. It would appear to us that the Portex laryngeal mask might need some design modifications to be a real alternative to the standard LMA. PMID- 15535498 TI - Night-shift discharge from intensive care unit increases the mortality-risk of ICU survivors. AB - Intensive Care (ICU) survivors discharged from ICU to the general ward at night have a higher mortality. We sought to clarify which factors, including night shift discharge, influence outcome following ICU discharge in a metropolitan hospital, using a cohort study of critically-ill patients between 1/1/1999 30/4/2003. Patients were excluded from analysis if they (a) died in ICU, (b) were transferred to another hospital, (c) had an ICU length of stay <8 hours, or (d) age <16 years. Logistic regression was used to derive a predictive model based on the following variables: patient demographics, severity of illness following ICU admission (APACHE II mortality-risk, p(m)), final diagnosis, discharge timing including premature or delayed (>4 hours) ICU discharge, and "limitation of medical treatment" orders. The outcome measures were patient status at hospital discharge and ICU readmission rate. Of the 1870 ICU survivors, 92 (4.9%) died after discharge from ICU. Patients discharged to the ward during the night-shift (2200-0730 hours) had a higher APACHE II score and crude mortality. The difference in APACHE II p(m) did not reach statistical significance. No significant calendar or seasonal pattern was identified. Logistic regression identified night-shift discharge (RR=1.7; 95% CI 1.03-2.9; P=0.03), limited medical treatment order (RR=5.1; 95% CI 2.2-12) and admission APACHE II p(m) (RR=3.3; 95% CI 1.3-7.6) as independent predictors of patient outcome following ICU transfer to the ward. CONCLUSION: At the time of ICU discharge to the ward three factors are predictive of hospital outcome: timing of ICU discharge, limited medical treatment orders and initial illness severity. PMID- 15535499 TI - Metropolitan audit of appropriate referrals refused admission to intensive care. AB - We undertook a three-month prospective cohort study of critically ill adult patients referred to the Intensive Care Units (ICUs) of public hospitals in metropolitan Melbourne and Geelong, Victoria. The aim was to ascertain the prevalence and immediate consequences of "refused" admission amongst patients appropriately referred to the ICU of first choice. Between August 1 and October 31, 1999, 10 (out of 12) public hospitals collected data. Three thousand and four patients were referred to these ICUs, and "refusals" were reported by all hospitals. A total of 282 (9.4%) patients were unable to be admitted to the ICU of first choice, giving a rate of 3.1 "refusals" per day. The reasons for "refusal" were limited staffing (52%) and shortage of beds (46%.) Acute inter hospital transfer (1.7 per day) was the most common immediate triage outcome (57%). These rates are higher than previously reported figures. We conclude that refused admission to the ICU of first choice, and acute inter-hospital transfer in this region and time period, were common events. PMID- 15535500 TI - Successful use of ECMO in adults with life-threatening infections. AB - Two cases of critically ill patients who received extracorporeal membrane oxygenation (ECMO) using different forms of circuitry and for different indications are presented. Both patients had life-threatening infections with septic shock and were not able to be supported by conventional means. The first patient had staphylococcal septicaemia and received venoarterial ECMO for circulatory failure. The second patient had psittacosis and received venovenous ECMO for respiratory failure. We discuss the expanding indications for this technology and the role it has to play in adult intensive care. PMID- 15535501 TI - Use of Sengstaken-Blakemore tube to stop massive upper gastrointestinal bleeding from Dieulafoy's lesion in the lower oesophagus. AB - Massive upper gastrointestinal bleeding from Dieulafoy's lesion in the gastrointestinal tract is uncommon. The use of the Sengstaken-Blakemore tube in acute gastrointestinal bleeding has become less common since endoscopic sclerotherapy and banding procedures have become widely available. The successful use of a Sengstaken-Blakemore tube to control acute massive upper gastrointestinal bleeding from a Dieulafoy's lesion in the lower oesophagus in an elderly man with severe coronary artery disease and heart failure is described. PMID- 15535502 TI - Rate-dependent left bundle branch block during anaesthesia. AB - Rate-dependent left bundle branch block during general anaesthesia is rare. Its occurrence makes electrocardiographic diagnosis of acute myocardial ischaemia or infarction difficult. It can also be confused with a slow rate ventricular tachycardia. We present a case of rate-dependent left bundle branch block in a patient with no previous history of ischaemic heart disease. Carotid sinus massage resulted in a decrease in heart rate and reversion to normal sinus rhythm. PMID- 15535503 TI - Safety warning with Datex-Ohmeda S/5 anaesthetic delivery unit design. PMID- 15535504 TI - The use of Storz bronchoscope in prevention of airway fire. PMID- 15535505 TI - Bispectral index monitoring. PMID- 15535506 TI - Awareness and hypoxia risk with Drager Cato and Fabius anaesthesia machines. PMID- 15535507 TI - Refractory hypotension in a patient chronically treated with a long acting angiotensin-converting enzyme inhibitor. PMID- 15535508 TI - Nitrous oxide in neonates and infants. PMID- 15535509 TI - Right of access to health information. AB - The access to health information is one of the rights of the user of the public health care system, but it is little respected in health services. This study investigates the problem from the perspective of the female clientele. A qualitative study was carried out with a sample of seventeen low-income mothers, which reside in the outskirts of the Federal District. The mothers are frequent users of the health care services, but their adherence to prescribed conducts depends on social and economic factors and how they perceive the various alternatives at their hand. Access to health information is hampered by the mother's low level of instruction, use of folklore medicine, faulty communication and lack of receptive/nurturing environment, indifference to social rights on the part of health providers. PMID- 15535510 TI - [Chilean's nursing knowledge organization and their tendencies]. AB - A Quantitative, descriptive and retrospective research that explored the Chilean Nursing knowledge organization an their tendencies. The universe was composed by scientists reports published in the Chilean nursing journals between 1965 and 2003 (N=214). Data were collected by an instrument based on the CIPE's and Nogueira's classification and cienciometria indications. Statistics measures of central tendency analysis was managed with SPSS. Some of the results obtained were: the more frequent study subject's was the professional nursing and the tendency is to be focused at the mature people's health necessities and their risk of being ill. It was found: little theoretical nursing sustenance at the reports. The "Ciencia y Enfermeria" is the journal that exhibits the best scientific quality of alls. PMID- 15535511 TI - [Items sterilized in humid heat: validation of the storage system]. AB - We aimed at identifying the period of validity of the sterilization of items by humid heat, taking into account the conditions of sterilization and storage. The validation was carried out by means of microbiological tests performed with instruments in the same load and evaluated in 0, 7, 10, 15, and 25 days. 30% of the instruments in each package were analysed and put in a Mueller Hinton solution and incubated at 37 degrees C for 72 hours. The reading was accomplished taking into consideration how dark the solution was. Out of the samples evaluated in nine loads, none showed microbiological growth. It was concluded that, although sterilization and storage conditions of the material were not fully in accordance with parameters recommended by the literature, the sterilization happened and was maintained for a period of 25 days. PMID- 15535512 TI - [Breast cancer: how women think and what they do]. AB - Changes brought about by the diagnosis and treatment of cancer always cause psychological, social, cultural, economic and spiritual disorders. We have tried to understand the significance of breast cancer for women and to find out what they do in order to detect it in an early stage. We have randomly chosen thirty four women who went to a cancer prevention centre in the town of Fortaleza. We have carried out clinical examinations of the breasts, taught the circular self examination technique and observed how the technique was learnt. We have analysed the results by means of an interactionist approach, and they have been organised according to their significance and to the development of early detection actions. We have concluded that women perceive cancer as a possibility of loss and death and make associations between people or past events and real or imaginary causes. Some of them perform early detection procedures, but the lack of information on the disease is evident, which makes it unfeasible to control breast cancer. PMID- 15535513 TI - [Knowledge and practice of breast self-examination by nurses]. AB - A qualitative study, with eleven nurses studying obstetric nursing in Teresina PI, the objectives of which were to describe their knowledge and practice of measures to control breast cancer and to discuss the practice of self-examination by nurses. A focus group was used as technique. The results show death or mutilations, followed by a bad prognosis, such as significances associated with breast cancer. They mentioned control measures such as healthy habits and examinations, especially self-examination, mammography, ultrasonography, clinical, static and dynamic examination, and biopsy in the confirmation of malignancy. Among the guidelines provided to customers, self-examination, healthy food habits, physical activity and no addictions were the most important ones. It can be concluded that these professionals did not perform self-examination very often, but the majority had healthy food habits. PMID- 15535514 TI - [Popular knowledge and health education in nursing training]. AB - From the point of view of including the health reform in the pedagogical projects of nursing courses, in accordance with curriculum guidelines, it contextualises the formation of a nurse for health education and the importance of bringing practice and care together, as well as includes the results of research with nurses in seven towns in rural communities in the state of Rio Grande do Sul. It is a qualitative survey with data collection by means of participatory observation, interviews, and document-based content analysis. These data were obtained in activities observed in the work of nurses in health education. It concludes that the work of nurses was full of educational actions, both collective and individual ones. On the other hand, undergraduate courses do not sufficiently prepare students to perform this role. PMID- 15535515 TI - [Diagnoses of fear and anxiety: content validation for a burned patient]. AB - This study aimed at identifying and performing content validation of the defining characteristics of the diagnoses of fear and anxiety for a burned patient. An instrument made up of manifestations of both diagnoses presented by Taxionomy I of the North American Nursing Diagnoses Association was elaborated, as well as others that were found in the literature. Sixteen nurses were interviewed. Out of the 97 characteristics presented in the instrument, 31 were excluded for the diagnoses "fear", and 27 for "anxiety". Eight characteristics had a score average above 0.70 for "fear" and 18 for "anxiety". The characteristics with the highest score were "expressed concern due to changes in life events" for anxiety and "insomnia referred or perceived" for fear. PMID- 15535516 TI - [Pain in children/adolescents with cancer: care dimensions]. AB - Focusing on the pain of a child/adolescent with cancer as a complex phenomenon, this study aims at describing the experiences of health professionals involved in caring for these patients in hospitalisations and analysing them, looking for a foundation in Edgar Morin's complexity ideas. Empirical data were collected in interviews, allowing for the visualisation of possibilities, such as a formation of fixed and integrated teams and the family's involvement in the care. Evidently, it is necessary to distinguish the professionals' multiple focuses and compose a care unit in which these focuses are articulated by means of a common project, to provide caring for the multiple dimensions of these patients' care. PMID- 15535517 TI - [The mother's role in breasfeeding her primiparous daughter: "the togetherness"]. AB - The authors' disquietudes are related to the structure for supporting women to breastfeed within their family environment. It is a qualitative study aiming at understanding the significance of breastfeeding among mothers and primiparous daughters, as well as identifying how the mother perceive herself as a means of support for her primiparous daughter and vice versa. The historic social construction of women for maternity has been used as a theoretical referential. The sample was made up of 10 women--five primiparous daughters and their mothers. The participation of the mother in her daughter's maternity was "to be along with her", sharing knowledge and life experiences. PMID- 15535518 TI - [People's participation in local health committees: showing lives, telling fights]. AB - This study aimed at analysing people's participation in Local Health Committees and understanding the participatory action of its members by identifying potentialities and limits in the process. We used the qualitative research approach, with interviews and observation as data collection methods. By applying Habermas's theoretical referential, we visualised the people's participation as a social and historical phenomenon marked by many fights. We found a communication that is very focused on strategic actions. However, we visualised possibilities to build an autonomous public space. We concluded that the people's participation can potentially act in the transformation of healthcare practices and, as such, it should be included by municipal administrators as a tool to elaborate and implement healthcare policies. PMID- 15535519 TI - [Perspectives for a new model of organisation in nursing]. AB - In this study, based on historical and dialectic materialism, I try to reflect on strategies to reorganize the work of nurses, aiming at minimizing the workers' process of physical and psychical fatigue. The collection of empirical data was carried out along with nurses of two health institutions in the south of the country, by means of interviews, observation and document-based analysis. Based on the indicatives, I present a model of organization inspired by the democracy of relationships, focusing on the workers' expression as multidimensional subjects, social actors, and not mere executors of delegated tasks, such as the social and technical division of work has established. A model aiming at establishing interpersonal relations that are more harmonic and horizontal and at sharing knowledge and tasks. PMID- 15535520 TI - [Prostitution in adolescence: interfaces with the family institution]. AB - Research carried out aiming at learning and analysing the context and significance of the dynamics in the family institution and its interfaces with the prostitution of adolescents. Data collection was carried out by means of life story interviews, and the content of the speeches underwent thematic analysis. We found a wide gap caused by the lack of dialogue and by how situations of abandonment, mother's death and life with family members being not a biological daughter of one of the parents have been experienced, breaking up with trust, respect and affect relationships, thus causing the loss of individual and collective identity. Violence within the family was identified through situations of sexual abuse, negligence and physical violence, thus making evident a true school of disrespect, abuse of power and gender/age hierarchy. PMID- 15535521 TI - [The significance of tubal ligation for people living in low-end neighborhoods in Porto Alegre]. AB - Health care access in the several social strata increases the demand for methods that ensure unflawed contraception. In this sense, the demand for tubal ligation by women older than 25 and with little schooling has increased. It is an exploratory and descriptive study with a qualitative approach aiming at: getting to know the significance of tubal ligation among sterilized women in the Vila Pinto, Porto Alegre; understanding the reasons for choosing this method and; finding out whether this surgical procedure entails any changes in sexual relationship. The participants in the study are 12 women who undertook tubal ligation within the period from July 2002 to June 2003. Data collection occurred by means of a semi-structured interview. The contents of the reports were analysed, wherefrom three topics emerged: decision to have tubal ligation; occurrence of tubal ligation and; experiences after tubal ligation. PMID- 15535522 TI - [Tendencies of life quality in the scientific nursing production]. AB - This study is characterized by bibliographical research of life quality, undertaken the BDENF and LILACS databases, at the BIREME Virtual Library. Nursing magazines rated as B and C international and B national were also researched by QUALIS/CAPES. The results obtained indicated that the production of knowledge on the topic being discussed is limited to situations of health problems, especially in specific groups. The production is still restricted to graduate courses, which limits the circulation among its peers. PMID- 15535523 TI - [The work of SUS (Unified Health System) nurses in Campinas: 1970s and 80s]. AB - We had the aim of analyzing the introduction and the practices developed by nurses in Health department of Campinas County in the period of 1978 and 1989. Among the practices developed by nurses we highlighted the supervision as instrument of evaluation/training and supervising/control of work; the training of aides for public health; the appointment and the nursing practices; the house calls; the ruling and developing of nursing technical procedures and the nursing registers. We reached the conclusion that there were transformations in the nursing working process, and a giant step took by them was very important for the establishment of the Public Health Network, the reorganization of model geared to health. PMID- 15535524 TI - [Analysis of nursing dissertations and theses on adolescence--Brazil, 1979-2000]. AB - This study aimed at checking the nursing knowledge produced about adolescence in Brazil from 1979 to 2000. The document-based research listed abstracts of dissertations and theses contained on the CD-ROM of the Centro de Estudos e Pesquisas em Enfermagem (CEPEN), the topics of which were adolescence/adolescent. The results, discussed in accordance with the concepts of Meleis (1997) and the relevant literature, show 46 works, 37 of them dissertations, defended in the Southeast Region (32), developed mostly between 1995 and 1999. The theoretical and methodological tendencies showed prevalence of clinical studies (22) related to the adolescent, of a qualitative nature (34). We concluded that there is a need for studies that include the adolescent in his/her family and community context. Also, greater incentive is necessary for the development of theses in poorer regions of the country. PMID- 15535525 TI - [Childhood cancer from the point of view the sick children's siblings: bibliographic review]. AB - The study aims at carrying out a bibliographic review of research on topics related to siblings of children with cancer; from 1998 to 2003, in order to identify the knowledge available in the field and provide subsidies for nursing care for siblings of children with cancer. We used the following databases: MEDLINE, LILACS, CINAHL, and PsycINFO. We also used full online articles, through the publishing house OVID, with the following keyword combinations: cancer and siblings; cancer, child and siblings; cancer, adolescent and siblings. The analysis of publications allowed the identification of four topics: the impact of the disease on healthy siblings; the significance of the disease for the healthy sibling; social support and care and intervention needs. The data indicated that research focusing on siblings of children with cancer has been growing, especially in the fields of nursing and psychology. They also show the concern with transforming clinical practice in order to effectively intervene in the care provided to the siblings' psychosocial needs. PMID- 15535526 TI - [Contribuction of nursing theories to build knowledge in the area]. AB - In this essay some aspects concerning the phenomenon of knowledge and of the activity of knowing are taken into consideration, serving to the authors'purpose to point out the perspective from which they understand which have been the contribution of the nursing theories for the construction of the knowledge of the area. The nursing theories select, define and interrelate concepts representative of phenomena that are in the nursing domain of interest. To guarantee the understanding and the recognition of the meaning of the concepts included in these theoretical frameworks is a basic aspect, therefore it raises their potential of practical applicability, either in education, research or nursing care. PMID- 15535527 TI - [Bandage to fix a peripheral intravenous catheter: an integrative literature review]. AB - This integrative literature review aimed at identifying, in the national and international literature, from 1992 to 2002, the publications about bandages to fix a peripheral intravenous catheter and the implications of its usage. Seven scientific articles were identified in the Medline database by using the following keywords: peripheral catheterisation and bandage. The results showed that nurses made 85% of the publications; 42.8% used descriptive comparative research as their methodology and; 71.4% investigated the occurrence of complications. Two studies showed a decrease in the occurrence of complications during the usage of transparent bandage coupled with a safety device. It was concluded that there is not enough basis to subsidise the selection of a bandage to fix a peripheral intravenous catheter. PMID- 15535528 TI - [Communicative action for building nursing knowledge]. AB - It is a theoretical essay aiming at proposing the usage of the communicative action theory, by philosopher Jurger Habermas, as a mediator of educational technologies in building nursing knowledge. Based on the contributions by the philosopher and authors such as Demo, Morin, Assman, Perrenoud, Waldow, and Ramos, we reflect on technological advances and the necessary innovations in the field of nursing education. We believe that this proposal can contribute to implementing the national curriculum guidelines in undergraduate courses. PMID- 15535529 TI - [Education as a means to face challenges posed to the elderly]. AB - This article discusses education as a means to face challenges imposed to the elderly by age and by society, thus allowing them to acquire new knowledge and obtain new opportunities to achieve physical and emotional well-being. Educational programmes for the elderly try to address these needs by working with several pedagogical procedures in order to awaken critical awareness in the search for a successful ageing. By means of continued education, these programmes have allowed the elderly to get up-to-date, acquire knowledge and participate in cultural, social, political, and leisure activities. PMID- 15535530 TI - [Educating a nurse for basic health care]. AB - This work aims at describing the experience of the 1st grade class in the Nursing Course at the Faculty of Medicine of Marilia (FAMEMA) in 2002. In 1998, a new Political-Pedagogical Project (PPP) was started aiming at educating a critical and reflexive professional, capable of teamwork, by promoting nursing care in a humane way. An integrated curriculum and the problematisation methodology were adopted as strategies to implement the PPP, tailoring education to the job market. Among the competencies to be developed toward basic health care, they employed a strategy to integrate 1st and 2nd grade classes in a microarea by applying the MAPP technique. PMID- 15535531 TI - [Nursing care of child: short retrospection]. AB - This study focuses on the overview of the childhood health policy, highlighted in the context of a hospital, in the city of Rio de Janeiro, aiming at describing the nursing assistance provided to children in hospitals in the first half of the century. It is a historical study, the primary written sources of which correspond to articles published in the Revista Brasileira de Enfermagem during the period under scrutiny. The secondary sources are made up of literature relevant to the topic. In the beginning of the 20th century, health care loses its social and philanthropic character to be subsidized by the government. Concerning childhood assistance, numerous hospitals opened in the 30's and 40's, even though the national healthcare policy was based on preventive actions. It may be concluded that childhood assistance up to the 50's was characterized by technical and scientific transformations in the hospital scenario, encouraging the construction of nursing knowledge in the area of paediatrics. PMID- 15535532 TI - It's survey time again! PMID- 15535533 TI - The PARIHS framework--a framework for guiding the implementation of evidence based practice. PMID- 15535534 TI - Baccalaureate nursing students' perceptions of the roles of registered nurses in the promotion of healthy aging. PMID- 15535535 TI - Web-based nurse executive dashboard. PMID- 15535536 TI - The first national reports on United States healthcare quality and disparities. AB - In the Healthcare Research and Quality Act of 1999 (Public Law 106-129), Congress mandated that the Agency for Healthcare Research and Quality (AHRQ) produce annual reports on healthcare quality and disparities in the United States. The National Healthcare Quality Report and the National Healthcare Disparities Report were first released in 2003 by the AHRQ. These reports include broad sets of performance measures to portray the nation's progress toward improving the quality of care provided to all Americans. This article provides an overview of the framework, development, and future uses of the reports by consumers, practitioners, researchers, and policy makers. PMID- 15535537 TI - Patient satisfaction and pain management: an educational approach. AB - The importance of assessing and managing pain has become paramount in today's hospital environment. Poor pain management is associated with impaired health, decreased patient satisfaction, and increased healthcare costs. This quality improvement project on an internal medicine unit at an urban teaching hospital examined the impact of pain education on patient satisfaction with pain management. Although pain scores did not improve, there were improvements made with respect to patient assessment, patient satisfaction, and nursing knowledge. PMID- 15535538 TI - Ventilator-associated pneumonia: improved clinical outcomes. AB - Nosocomial pneumonia is the second most costly infection occurring in US hospitals. It is associated with high mortality and morbidity and is considered one of the most difficult infections to diagnose and prevent. In 1999, St Joseph Hospital formed a multidisciplinary Ventilator-Associated Pneumonia Performance Improvement Team to tackle the myriad of issues involved in the prevention of ventilator-associated pneumonia and the care of this high-risk patient population. The accomplishments of the team were significant, with a 95% reduction in ventilator-associated pneumonia. PMID- 15535539 TI - Quality focus of the first line nurse manager and relationship to unit outcomes. AB - The purpose of this article was to examine the relationship between the quality focus of the first line nurse manager and patient satisfaction, job satisfaction of the nursing personnel, unit effectiveness, staff perceptions of quality, and nursing personnel turnover. Of the unit outcomes that were tested, quality focus was found to be a significant predictor variable only for staff nurse job satisfaction (R2 = 0.218). Managers need to be aware of their vision for quality of patient care and how this translates into expectations for nursing care quality and overall quality of the organization. PMID- 15535540 TI - Snoezelen activity: the Good Shepherd Nursing Home experience. AB - Care of the resident with dementia can be both challenging and unpredictable. Activities provided for nursing home residents often have rules and may be a source of frustration for residents with advancing dementia. Snoezelen, or multisensory therapy, offers a failure-free activity in an enabling environment that can both stimulate and relax the resident with dementia. Good Shepherd Nursing Home in Versailles, Mo, undertook a 1-year outcome-based quality improvement project to find if use of Snoezelen therapy could reduce the number of behavioral symptoms that residents were suffering from. While there are still barriers to the use Snoezelen therapy, employees at Good Shepherd Nursing Home believe that the use of Snoezelen therapy has been a successful and rewarding experience for both residents and staff members. PMID- 15535541 TI - Nonpharmacological strategies for improving heart failure outcomes in the community: a systematic review. AB - The purpose of this systematic review was to synthesize current evidence regarding nonpharmacological approaches to heart failure (HF) management. Following a literature search, identified studies were coded using the Heart Failure Study Assessment Scale (HFSAS) developed by the authors. Results included high-quality ratings, a predominance of multidisciplinary disease management studies, decreased readmission rates, and some improvement in quality of life. Implications for practice, leadership, education, and research are described. PMID- 15535542 TI - Down the rabbit hole: examining outcomes of nurse midwifery care. AB - Care of the laboring woman and subsequent birth interventions have generally been based on tradition rather than a systematic examination of the cost-effectiveness of the interventions. This retrospective study examined the outcomes of nurse midwifery care at a large metropolitan university clinic setting. Findings suggest that more sensitive cost and quality indicators of nurse midwifery care need to be developed, and the effect of these on outcomes needs to be elucidated. PMID- 15535543 TI - Measuring healthcare outcomes to improve quality of care across post--acute care provider settings. AB - Post-acute care (PAC) occurs in a variety of settings-skilled nursing facilities (nursing homes), rehabilitation facilities, and home health agencies. To evaluate the impact of care processes on clinical outcomes and implement changes designed to improve outcomes, one must begin by measuring outcomes in a valid, reliable manner that allows for comparisons to reference or benchmarking data. Currently, several data sets exist in PAC settings for the purpose of outcome measurement. However, there is a need for comparable information across settings to ensure the quality and continuity of care. This article reviews various existing data sets used in PAC settings, examines ongoing projects to create a single set of measures, and suggests some directions for future research. PMID- 15535544 TI - Molecule-to-metal bonds: electrografting polymers on conducting surfaces. AB - Electrografting is a powerful and versatile technique for modifying and decorating conducting surfaces with organic matter. Mainly based on the electro induced polymerization of dissolved electro-active monomers on metallic or semiconducting surfaces, it finds applications in various fields including biocompatibility, protection against corrosion, lubrication, soldering, functionalization, adhesion, and template chemistry. Starting from experimental observations, this Review highlights the mechanism of the formation of covalent metal-carbon bonds by electro-induced processes, together with major applications such as derivatization of conducting surfaces with biomolecules that can be used in biosensing, lubrication of low-level electrical contacts, reversible trapping of ionic waste on reactive electrografted surfaces as an alternative to ion exchange resins, and localized modification of conducting surfaces, a one-step process providing submicrometer grafted areas and which is used in microelectronics. PMID- 15535545 TI - Nanotechnology and single molecules. PMID- 15535546 TI - Pulsed-laser production and detection of spin-polarized hydrogen atoms. AB - Spin-polarized hydrogen (SPH) atoms have traditionally been produced and detected using complex experimental methods with poor time resolution. Recently, SPH has been produced by pulsed-laser photodissociation of HCl using circularly polarized light. In combination with the proposed detection of SPH via polarized fluorescence, this approach should allow the production and spatially resolved detection of SPH with a higher sensitivity than that currently available, and with a time resolution in the nanosecond regime. This represents an improvement of several orders of magnitude over the existing methods. PMID- 15535547 TI - Protonated water clusters: the third dimension. PMID- 15535548 TI - The Jahn-Teller effect of the TiIII ion in aqueous solution: extended ab initio QM/MM molecular dynamics simulations. AB - Combined ab initio quantum mechanical/molecular mechanical (QM/MM) molecular dynamics simulations, including only the first and the first and second hydration shells in the QM region, were performed for TiIII in aqueous solution. The hydration structure of TiIII is discussed in terms of radial distribution functions, coordination-number distributions and several angle distributions. Dynamical properties, such as librational and vibrational motions and TiIII-O vibrations, were evaluated. A fast dynamical Jahn-Teller effect of TiIII(aq) was observed in the QM/MM simulations, in particular when the second hydration shell was included into the QM region. The results justify the computational effort required for the inclusion of the second hydration shell into the QM region and show the importance of this effort for obtaining accurate hydration-shell geometries, dynamical properties, and details of the Jahn-Teller effect. PMID- 15535549 TI - Two-photon dissociation spectroscopy of state-selected HCl+ and DCl+ ions. AB - HCl+ and DCl+ ions were formed via the R(1) pump line of the f3delta2(v'=0)<- sigma+(v''=0) REMPI process. For these ions, the two-photon dissociation spectroscopy, resonance-enhanced via the A2sigma+(v')<--pi3/2(v''=0) transition, was investigated for various intermediate states of HCl+ (v'=4,5,6) and DCl+ (v'=6,7,8,9). From the analysis of the data, spectroscopic parameters of the X and the A states were derived (including the lambda-doubling in the X state and the spin-rotation coupling in the A state). Some of the parameters deviate considerably from literature data. The spectra provide clear evidence that the REMPI process employed for forming the ions has a very high rotational selectivity. PMID- 15535550 TI - Accurate pKa determination for a heterogeneous group of organic molecules. AB - Single-molecule studies that allow to compute pKa values, proton affinities (gas phase acidity/basicity) and the electrostatic energy of solvation have been performed for a heterogeneous set of 26 organic compounds. Quantum mechanical density functional theory (DFT) using the Becke-half&half and B3LYP functionals on optimized molecular geometries have been carried out to investigate the energetics of gas-phase protonation. The electrostatic contribution to the solvation energies of protonated and deprotonated compounds were calculated by solving the Poisson equation using atomic charges generated by fitting the electrostatic potential derived from the molecular wave functions in vacuum. The combination of gas-phase and electrostatic solvation energies by means of the thermodynamic cycle enabled us to compute pKa values for the 26 compounds, which cover six distinct chemical groups (carboxylic acids, benzoic acids, phenols, imides, pyridines and imidazoles). The computational procedure for determining pKa values is accurate and transferable with a root-mean-square deviation of 0.53 and 0.57 pKa units and a maximum error of 1.0 pKa and 1.3 pKa units for Becke half&half and B3LYP DFT functionals, respectively. PMID- 15535551 TI - Photon counting histogram: one-photon excitation. AB - The photon counting histogram (PCH) analysis is a fluorescence fluctuation method that is able to characterize the brightness and concentration of different fluorescent species present in a liquid sample. We find that the PCH model using a three-dimensional Gaussian observation volume profile is inadequate for fitting experimental data obtained from a confocal setup with one-photon excitation. We propose an imoroved model, which is based on the correction to the observation volume profile for the out-of-focus emission. We demonstrate that this model is able to resolve different species present under a wide range of conditions. Attention is given to how this model allows the examination of the effects of different instrumental setups on the resolvability. PMID- 15535552 TI - Determination of the heterogeneous association constants of metal ions to omega mercaptoalkanoic acids by using double-layer capacity measurements. AB - The binding of metal ions to ligands in homogeneous solutions and that to the same ligands anchored to metallic surfaces through self-assembled monolayers (SAMs) are expected to differ primarily due to the difference in the degree of freedom of the ligands and the surface potential. We studied the heterogeneous binding of CdII ions to omega-mercaptoalkanoic-acid SAMs on Au. This was accomplished by adding metal ions at a constant pH and following the changes in the double-layer capacity. A mathematical treatment, which is based on calculating the electrochemical-potential differences at the double layer solution interface, has been developed. Our approach follows that proposed by White et al. and Kakiuchi, who used the acid-base equilibrium at the monolayer electrolyte interface as a means of calculating the pK of ionizable SAMs. Experimentally, SAMs of omega-mercaptoalkanoic acids, HS(CH2)nCO2H, with different chain lengths (i.e., n=2, 5, and 10) in 0.1 M sodium perchlorate were assembled on Au surfaces and studied. The capacity was measured first in the absence of CdII at different pH values, and then at a constant pH while increasing the concentration of CdII in the solution. We found that the interfacial capacity decreased as the concentration (of either protons or CdII) increased. The results matched the model fairly well, which allowed the extraction of the thermodynamic equilibrium constant that is established at the monolayer-electrolyte interface. The suggested mathematical treatment of this model system is simple and yet very useful for estimating the heterogeneous association constants of metal ions by SAMs. PMID- 15535553 TI - Gaseous ion activation dynamics: the role of the bulk gas in the racemization of chiral oxonium ions. AB - The kinetics of the inversion of configuration of a family of chiral oxonium ions, that is, O-protonated 1-aryl-1-methoxyethanes [YMe+], were investigated in two different gaseous media (in CH3X with X=F and X=Cl) at 720 torr of pressure and in the temperature range: 25-140 degrees C. The activation parameters of the [YMe+] inversion reaction were found to obey two different isokinetic relationships (IKR), depending on the nature and the position of the substituents in the oxonium ions and on the nature of the bulk gas employed. The observation of two IKR for the same family of reactions was related to a switchover in the resonant vibrational energy exchange between the reactants' critical mode, active in the transition state (omega), and the discrete vibrational levels v of the bulk gas. In CH3F, this vibrational-vibrational coupling switchover concerns the out-of-plane C-F...H-O bending) the phi family) and the H3C-F stretching (the gamma family) modes in the proton-bound [CH3F.YMe+] complex. In CH3Cl, the coupling switchover concerns the out-of-plane C-Cl...H-O bending (the phi family) and the H3C-Cl methyl group rocking (the gamma family) modes in the proton-bound [CH3Cl.YMe+] complex. The [YMe+] activation dynamics also determine the inversion dynamics. The [YMe+]ret<==>[YMe+]inv isomerization for the phi family involves the same "thermodynamically most favorable" transition state in both the CH3F and the CH3Cl media, whereas the same process for the gamma family proceeds through different, dynamically favored transition states. PMID- 15535554 TI - Revision of the dissociation energies of mercury chalcogenides--unusual types of mercury bonding. AB - Mercury chalcogenides HgE (E=O, S, Se, etc.) are described in the literature to possess rather stable bonds with bond dissociation energies between 53 and 30 kcal mol(-1), which is actually difficult to understand in view of the closed shell electron configuration of the Hg atom in its ground state (...4f(14)5d(10)6s(2)). Based on relativistically corrected many body perturbation theory and coupled-cluster theory [IORAmm/MP4, Feenberg-scaled IORAmm/MP4, IORAmm/CCSD(T)] in connection with IORAmm/B3LYP theory and a [17s14p9d5f]/aug-cc-pVTZ basis set, it is shown that the covalent HgE bond is rather weak (2-7 kcal mol(-1)), the ground state of HgE is a triplet rather than a singlet state, and that the experimental bond dissociation energies have been obtained for dimers (or mixtures of monomers, dimers, and even trimers) Hg2E2 rather than true monomers. The dimers possess association energies of more than 100 kcal mol(-1) due to electrostatic forces between the monomer units. The covalent bond between Hg and E is in so far peculiar as it requires a charge transfer from Hg to E (depending on the electronegativity of E) for the creation of a single bond, which is supported by electrostatic forces. However, a bonding between Hg and E is reduced by strong lone pair-lone pair repulsion to a couple of kcal mol(-1). Since a triplet configuration possesses somewhat lower destabilizing lone pair energies, the triplet state is more stable. In the dimer, there is a Hg-Hg pi bond of bond order 0.66 without any a support. Weak covalent Hg-O interactions are supported by electrostatic bonding. The results for the mercury chalcogenides suggests that all experimental dissociation energies for group-12 chalcogenides have to be revised because of erroneous measurements. PMID- 15535555 TI - Azulene-to-naphthalene rearrangement: the Car-Parrinello metadynamics method explores various reaction mechanisms. AB - We studied the thermal intramolecular and radical rearrangement of azulene to naphthalene by employing a novel metadynamics method based on Car-Parrinello molecular dynamics. We demonstrate that relatively short simulations can provide us with several possible reaction mechanisms for the rearrangement. We show that different choices of the collective coordinates can steer the reaction along different pathways, thus offering the possibility of choosing the most probable mechanism. We consider herein three intramolecular mechanisms and two radical pathways. We found the norcaradiene pathway to be the preferable intramolecular mechanism, whereas the spiran mechanism is the favored radical route. We obtained high activation energies for all the intramolecular pathways (81.5-98.6 kcal mol( 1)), whereas the radical routes have activation energies of 24-39 kcal mol(-1). The calculations have also resulted in elementary steps and intermediates not yet considered. A few attractive features of the metadynamics method in studying chemical reactions are pointed out. PMID- 15535556 TI - On the quantum nature of an excess proton in liquid hydrogen fluoride. AB - Liquid hydrogen fluoride consists of chains of hydrogen-bonded molecules. The nature of an excess proton in liquid HF, which is of interest not only for its own sake, but also in relation to super-acid chemistry and to its behavior in water, has been studied using computer simulations. The methodology employed is the density-functional-theory-based path-integral Car-Parrinello ab initio molecular dynamics. The excess proton, which formally exists as a H2F+ or a H2F2+ defect in an HF chain, is found to strongly perturb the chain to which it is attached. Moreover, due to large zero-point energy, the charge defect is largely delocalized over several HF molecules. PMID- 15535557 TI - Experimental and theoretical study of the photophysics and structures of europium cryptates incorporating 3,3'-bi-isoquinoline-2,2'-dioxide. AB - A detailed photophysical study of [Eu within (biqO2.2.2)(CF3SO3)](CF3SO3)2. CH3CN.H2O (Eu within 1) and two other types of cryptates incorporating three 3,3' biisoquinoline-2,2'-dioxide units has been performed. Structural crystallographic data of Eu within 1, electronic structure calculations and theoretical models were used to obtain the intramolecular energy transfer rates and the appropriate set of rate equations, which was solved numerically. Quantum yields and decay lifetimes were obtained from these results and compared to the experimental data. The role of the ligand-to-metal charge transfer (LMCT) states was ascertained. A theoretical ligand field and intensity analysis was carried out and the results agree very well with the emission spectra. The molecular structures of the lanthanide cryptates were successfully modelled by the YIII ion using the restricted Hartree-Fock (RHF) method, with the advantage of dealing with closed shell systems. These molecular structures were used to explain the drastic differences in the photophysics of the three EuIII cryptates. PMID- 15535558 TI - Visual observation of contact-induced intercrystalline migration of aromatic species adsorbed in zeolites by fluorescence microscopy. AB - Intercrystalline migration and a migration-assisted chemical reaction of adsorbed aromatic species between zeolite particles in physical contact were visualized by fluorescence microscopy coupled with a particle manipulation technique. The luminescence color characteristics of particular zeolite particles originating from the specific photochemistry of adsorbed species was exploited to follow the migration of the molecules. Two examples are shown that are relevant to the visualization of the time-dependent migration process: A one guest-two sets of zeolite crystals system: chrysene (Chry)-loaded zeolite Na+ -X (the sodium form of zeolite X) crystals were placed in contact with unloaded Tl+ -X (thallium exchanged X) crystals and allowed to stand at room temperature. Initially, the blue fluorescence of Chry was detected only from the Na+ -X particles, but later, the development of green phosphorescence emission was discernible from the Tl+ -X which suggests that Chry migrated from the Na+ -X to the Tl+ -X crystals. A two guest-species systems: Electron-donating Chry-loaded Na+ -X crystals were placed in contact with electron-accepting 1,2,4,5-tetracyanobenzene (TCNB)-loaded Na+ -X or Na+ -Y crystals. With time, the former system (Chry/Na+ -X and TCNB/Na+ -X) gave rise to the emission of Chry-TCNB charge-transfer complexes resulting mainly from the migration of Chry while the latter system (Chry/Na+ -X and TCNB/Na+ -Y) afforded the same emission resulting largely from the migration of TCNB. The present investigation reveals that there is a certain direction for guest migration depending on the zeolite host and the nature of host-guest or guest guest interaction. PMID- 15535559 TI - Monolayers of zeolite A containing luminescent silver sulfide clusters. PMID- 15535560 TI - Diffusion in protein crystals--a computer simulation. PMID- 15535561 TI - In-situ observation of nanowire growth from luminescent CdTe nanocrystals in a phosphate buffer solution. PMID- 15535562 TI - Double-system-size resonance for spiking activity of coupled Hodgkin-Huxley neurons. PMID- 15535563 TI - Anisotropic translational diffusion of single fluorescent perylene molecules in a nematic liquid crystal. PMID- 15535564 TI - Size-dependent spectroscopic properties and thermochromic behavior in poly(substituted thiophene) nanoparticles. PMID- 15535565 TI - Low-pressure organometallic chemical vapor deposition of indium nitride on titanium dioxide nanoparticles. PMID- 15535566 TI - A new cubic phase containing DNA and a surfactant. PMID- 15535567 TI - Intramolecular electrostatics: Coulomb's law at sub-nanometers. PMID- 15535568 TI - Comparison of sample units for estimating population abundance and rates of change of adult horn fly (Diptera: Muscidae). AB - This study compared the reliability of population estimates of adult horn fly, Haematobia irritans (L.), obtained using different sample units. Mean-variance relationships were similar for abundance estimates obtained by counting flies on the sunny sides of cattle, on the upper body, and on the whole animal. Precision varied among the sample units, and was lowest for estimates obtained using the sunny side. Abundance estimates obtained using the sunny side and upper body sample units were related to estimates obtained using the whole body sample unit. However, the proportion of flies in the upper body and sunny side sample units declined with increasing fly density. Seasonal movement toward the belly accounted for this decline. This movement resulted in bias in estimating rates of change based on counting flies on the sunny side and upper body sample units. Rates of change based on sampling the sunny side were more biased than estimates based on the upper body sampling unit. Bias in estimating rates of change was examined using an analytical model compared with field data, and resulted from changes in the proportion of flies occupying the sample unit. Bias also increased with increasing actual rates of change. The implication of these findings for studying horn fly populations are discussed. PMID- 15535569 TI - Evaluation of Stomoxys calcitrans (Diptera: Muscidae) behavioral response to human and related odors in a triple cage olfactometer with insect traps. AB - A triple cage olfactometer provided with insect traps was used for evaluating behavioral responses of Stomoxys calcitrans (L.) females to human skin and breath, CO2, and L-lactic acid analogs. After demonstrating there were no significant differences caused by cage location or time of day, 3 sets of 3 olfactometer tests were performed in a day, every 2 h beginning at 0900 hours. When a human hand was used as attractant, the attraction (expressed as percentage of trapped flies) increased as a function of the time; an inverted U-shaped relationship between attractancy and air speed was observed; and variation in fly density in the range 25-75 per cage did not affect the attraction response. When human breath was used as attractant the attraction increased linearly as a function of time and it was exhalation frequency dependent; when air flow was absent the highest response was observed; and 24- to 38-h-old flies were more attracted than younger and older. When CO2 was tested, activation and orientation and probing behavior were concentration dependent with flows ranging between 0.0001 and 0.038 liter s(-1), but attraction was not. No attraction was observed with 10, 100, or 1,000 microg of compounds related to L-lactic acid and several synthetic human odors and related compounds, although orientation was often observed. PMID- 15535570 TI - Oviposition and maintenance of Forcipomyia (Lasiohelea) townsvillensis (Diptera: Ceratopogonidae) in the laboratory. AB - Fecundity, oogenesis, oviposition, and percentage egg hatch were quantified for the blood-feeding midge Forcipomyia (Lasiohelea) townsvillensis (Taylor). Data are similar to that reported for other species of blood-feeding Forcipomyia. Eggs rarely developed from a partial blood meal but invariably developed after a single, complete blood meal. Results suggest that this species is anautogenous. Oviposition media were investigated and a successful medium and holding chamber type identified. Longevity of adults in the laboratory was studied and indicates the possibility for >1 gonotrophic cycle to occur. Adult survival at different relative humidities showed midges can survive 35-98% RH. Rearing of larvae in the laboratory and culture media are discussed. The data supplied in this paper provide the basis for the laboratory culture of F. (L.) townsvillensis. PMID- 15535571 TI - Photosynthesis modulates the plant feeding of Phlebotomus papatasi (Diptera: Psychodidae). AB - Phlebotomus papatasi (Scopoli), the vector of Leishmania major (Yakimoff & Schokhor), feeds on plants in desert habitats in the Jordan Valley. At the end of the dry summer, the life span of sand flies is short and the amount of sugars in their guts is small. In this season the plants are under the stress of heat and dehydration. This stress arrests the photosynthesis and decreases the amounts of the main end products, sucrose and starch. We presumed that the paucity of sugars in the sand fly plant tissue diet resulted from the arrest of photosynthesis. To test this assumption, we compared the feeding of sand flies on branches of Capparis spinosa (L.) that had been kept for 24 h in darkness and on branches cut after a normal day of photosynthesis. In darkness, the branches had lost more than half of their sugar content. Afterward they were fed upon overnight by 45.2% of female and 14.3% of male sand flies. A higher proportion of 81.0% females and 38.7% males fed on branches from natural conditions and these fed flies were significantly heavier. Laboratory experiments also showed that plant tissue meals of P. papatasi often include starch grains. Such grains were found also in 50% of field-caught males and females. The nutritive potential of plant tissues was demonstrated by the 33-d median survival of P. papatasi series that had been maintained on fresh C. spinosa branches and water. PMID- 15535572 TI - Genetic divergence among Venezuelan populations of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae). AB - Lutzomyia longipalpis (Lutz & Neiva) is the primary vector of visceral leishmaniasis in Venezuela. An analysis of alleles at seven enzyme-encoding loci among four populations from different geographic and epidemiological regions revealed strong genetic substructuring. Isozyme analysis indicated that L. longipalpis in Venezuela is a complex of at least two subspecies. Possible differences in population size during their evolutionary histories, varying colonization histories and geological events may explain discrepancies in the patterns of variation observed at genetic markers between these four populations. PMID- 15535573 TI - Susceptibility of three laboratory strains of Anopheles albimanus (Diptera: Culicidae) to coindigenous Plasmodium vivax circumsporozoite protein phenotypes in southern Mexico. AB - The susceptibility to two coindigenous Plasmodium vivax Grassi & Feletti phenotypes VK210 and VK247 of three colonized Anopheles albimanus Wiedemann strains (white-striped, green and brown) from southern Mexico was investigated. Mosquitoes of the three strains were simultaneously fed with P. vivax-infected patient blood and examined 1 wk later for the presence of oocysts. The circumsporozoite protein phenotype type (VK210 and VK247) was determined by immunoflorescence of salivary gland sporozoites using monoclonal antibodies. The proportions of specimens infected and the number of oocyst per mosquito indicated that all mosquito strains were more susceptible to the phenotype VK210 than to VK247, but the white-striped strain was more susceptible to both parasite phenotypes than the other two strains. PMID- 15535574 TI - Methodology for sampling questing nymphs of Ixodes ricinus (Acari: Ixodidae), the principal vector of Lyme disease in Europe. AB - To assess the Lyme borreliosis vector population density we set up a methodology for sampling the Ixodes ricinus L. population host questing on the vegetation. We focused on the collection of the nymphal stage, which is the principal stage of disease transmission to humans. This study was carried out in Rambouillet forest (Yvelines, France) where seven study areas were demarcated. These areas are maximally homogeneous for plant species using a finer scale than the phytosociological classification as defined by the method of landscape diagnostics. Out of 23 collections performed from March 1997 to May 1998, 2,906 I. ricinus nymphs were collected. The sampling technique chosen was the cloth lure technique. The technical parameters were studied and fixed (cloth type, cloth size, sample size, researcher position). It appeared that toweling was the best cloth type to optimize the number of ticks collected; the position of the researcher had no effect on tick samples. To satisfy the criteria for correct sampling, we studied representativity, randomness, and nonselectivity of our methodology. The spatial distribution of nymphs in a homogeneous area was close to random and thus very few subsamples were needed to obtain a relative density which was representative. No significant differences were found between random samples and following transect samples; and nonselectivity was totally satisfied because we only worked on questing nymphs. We grouped the samples that presented no significant differences to attribute a density index, which varied from 0 to 5. This methodology, applied with the same parameters, offers potential for producing comparable results from studies in different geographical areas and at different times of the years. PMID- 15535575 TI - Distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes (Diptera: Culicidae). AB - Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are associated with a variety of reproductive alterations in their hosts, the best known being cytoplasmic incompatability. The Wolbachia pipientis assemblage has been divided into two major groups (A and B) and 12 subgroups. We report herein the first systematic survey of Wolbachia in mosquitoes, and the first survey classifying Wolbachia infections by subgroup. Wolbachia were detected in 28.1% of 89 wild-caught mosquito species, based on a polymerase chain reaction assay using ftsZ and wsp gene primers. Infections were found in all major disease vector genera except Anopheles. Nine of the 12 Wolbachia subgroups were represented. Group B Wolbachia strains showed more phylogenetic concordance with their host taxa than group A strains. Of the 25 positive mosquito species, five were superinfected with group A bacteria strains (AA), eight were superinfected with A and B strains (AB), and one was superinfected with group B strains (BB). The widespread distribution of Wolbachia among mosquito species further supports their potential importance in the genetic control of disease vectors. PMID- 15535576 TI - Colonization of Phlebotomus neglectus (Diptera: Psychodidae), the major vector of visceral leishmaniasis in Greece. AB - Colonization of Phlebotomus neglectus Tonnoir, the major vector of visceral leishmaniasis, in Greece is reported for the first time. Starting with wild caught specimens, a small closed colony was established that was maintained for 17 mo or 10 generations. Gonotrophic discordance, stenogamic mating behavior, low fecundity, and dormancy because of low temperature were the most important findings that characterized the colony. PMID- 15535577 TI - Ehrlichia chaffeensis (Rickettsiales: Ehrlichieae) infection in Amblyomma americanum (Acari: Ixodidae) at Aberdeen Proving Ground, Maryland. AB - Human monocytic ehrlichiosis (HME) is a sometimes fatal, emerging tick-borne disease caused by the bacterium Ehrlichia chaffeensis. It is frequently misdiagnosed because its symptoms mimic those of the flu. Current evidence indicates that Amblyomma americanum (L.), the lone star tick, is the major vector of HME. To determine if E. chaffeensis is present in ticks at Aberdeen Proving Ground, MD, questing A. americanum ticks were collected from 33 sites. Nucleic acid was extracted from 34 adult and 81 nymphal pools. Sequences diagnostic for E. chaffeensis from three different loci (16S rRNA, 120-kDa protein, and a variable-length polymerase chain reaction [PCR] target, or VLPT) were targeted for amplification by the PCR. Fifty-two percent of the collection sites yielded pools infected with E. chaffeensis, confirming the presence and widespread distribution of E. chaffeensis at Aberdeen Proving Ground. Analysis with the both the 120-kDa protein primers and the VLPT primers showed that genetic variance exists. A novel combination of variance for the two loci was detected in two tick pools. The pathogenic implications of genetic variation in E. chaffeensis are as yet unknown. PMID- 15535578 TI - Estimating population size and drag sampling efficiency for the blacklegged tick (Acari: Ixodidae). AB - Estimates of absolute density were determined over a 5-yr period (1990-1994) for a population of Ixodes scapularis Say located in Westchester County, NY, by mark release-recapture (nymphs and adults) and removal (larvae) methods. Density estimates for larvae ranged from 5.2 to 16.5/m2 and averaged 11.5/m2. Values for nymphs varied as much as fourfold among successive years, ranging from 0.5 to 2.3/m2 and averaging 1.2/m2, whereas adult density ranged from 0.3 to 0.4/m2, averaging 0.33/m2. Natural mortality of nymphs and adults was measured in experimental cages during population estimation periods, and indicated that survival declined linearly over the short-term and did not significantly influence estimates. Drag sampling efficiency, the proportion of the estimated population obtained in a single sample, averaged 6.3% among all stages. Efficiency was not significantly different among stages and was independent of tick density within a given life stage. The population estimation techniques employed in this study are well suited for use with I. scapularis and can provide data that offer insights into mortality patterns in individual populations. PMID- 15535579 TI - Invertebrate carcasses as a resource for competing Aedes albopictus and Aedes aegypti (Diptera: Culicidae). AB - Terrestrial invertebrate carcasses are an important resource for insects developing in pitcher plants. However, little is known of the role of these carcasses in other containers, which also receive leaf fall and stemflow inputs. This experiment investigated effects of accumulated invertebrate carcasses as a resource for two competing mosquitoes, Aedes albopictus (Skuse) and Aedes aegypti (L.), whether either species differentially benefited from accumulated carcasses, and if such a benefit affected interspecific competition. First, we measured accumulation of invertebrate carcasses in standard containers at a field site. We then used a replacement series with five different species ratios at the same total density, and varied the input of invertebrate carcasses [dead Drosophila melanogaster (Meigen) ] in three levels: none, the average input from our field site, or the maximum input recorded at our field site. Survivorship, development time, and mass were measured for each mosquito species as correlates of population growth, and were used to calculate a population performance index, lambda'. There were strong positive effects of invertebrate carcass additions on all growth correlates and lambda'. Differences in performance between species were pronounced in small or no carcass additions and absent in large inputs of invertebrate carcasses, but there was little evidence that inputs of invertebrate carcasses altered the competitive advantage in this system. These results suggest that terrestrial invertebrate carcasses may be an important resource for many types of container communities, and large accumulations of dead invertebrates may reduce resource competition between these mosquitoes, thus favoring coexistence. We propose that the total amount of resource, including accumulated invertebrate carcasses, may explain observed patterns of replacement involving these mosquitoes. PMID- 15535580 TI - Influence of the blood meal source on the development of Triatoma infestans, Triatoma brasiliensis, Triatoma sordida, and Triatoma pseudomaculata (Heteroptera, Reduviidae). AB - The objective of this study was to determine the influence of the blood meal source on the life cycle and reproductive development of female Triatoma infestans (Klug, 1834), Triatoma brasiliensis Neiva, 1911, Triatoma sordida (Stal, 1859), and Triatoma pseudomaculata Corrga & Espfinola, 1964. In all triatomine species studied the life cycle was shorter for the groups fed on mice than for those fed on pigeons, the range of differences being between 1.5 times (T pseudomaculata and T. infestans) and 2.4 times (T brasiliensis). The mortality rate of nymphs during the life cycle tended to be greater in insects fed on pigeons than in those fed on mice, the differences for T. brasiliensis being statistically significant. Females of T sordida and T pseudomaculata had a greater fecundity than those of T. infestans and T. brasiliensis independently of the blood meal source. The differences of fecundity observed probably reflect differences in the availability of blood in the silvatic ecotopes of these species, meals being more frequent for T. infestans and T brasiliensis, which live at high densities in association with rodents in highly stable ecotopes. Because T. sordida and T. pseudomaculata live in more unstable ecotopes with fewer sources of blood they form small sparse colonies and invest more energy in reproduction than maintenance. PMID- 15535581 TI - Aedes notoscriptus (Diptera: Culicidae) survival and dispersal estimated by mark release-recapture in Brisbane, Queensland, Australia. AB - The survival and dispersal of adult Aedes notoscriptus (Skuse) were estimated in Brisbane, Queensland, Australia, where this species has been identified as a vector of Ross River and Barmah Forest viruses and shown to be orally susceptible to dengue virus types 1-4. Standard mark-release-recapture methods were used. Before the field study, laboratory trials showed that marking Ae. notoscriptus with fluorescent powder had no effect on survivorship. Female recapture rates of 10.6 and 2.3% over an 8-d period were achieved for two cohorts of marked Ae. notoscriptus released at the same field site. No males were recaptured over the 8 d period. The probability of daily survival was calculated using the Saul model as 0.77 and 0.79 for blue- and pink-marked females, respectively. For blue- and pink-marked females, the mean distance traveled was 105.2 and 179.9 m, and the maximum distance traveled was 195 and 238 m, respectively. The results indicate that protected harborage sites and shade influenced the distribution of Ae. notoscriptus within the study site. PMID- 15535582 TI - Life tables of Toxorhynchites rutilus (Diptera: Culicidae) in nature in southern Florida. AB - Stage-specific survivorship curves were constructed for the immature stages of a native, predatory mosquito, Toxorhynchites rutilus (Coquillet), by regular censuses in the summer and fall of water-holding treeholes and tires. Survival from egg to adult ranged from 1.8 to 5.6%, and survivorship patterns were significantly heterogeneous between seasons and container types. The probability of death was highest in the first and fourth larval instars. Rainfall and drought were relatively unimportant risk factors during this study, but the probabilities of disappearance of eggs and first and fourth instars were significantly higher in the presence of large conspecifics, suggesting that cannibalism is a major source of mortality. PMID- 15535583 TI - Effects of permethrin on the salivary glands and neuroendocrine organs of unfed female Hyalomma (Hyalomma) dromedarii (Ixodoidea: Ixodidae). AB - Permethrin-impregnated fabric has been shown to be an effective repellent against various tick species. However, some tick species are not repelled by this chemical. In Hyalomma dromedarii (Koch), permethrin exposure is reported to actually enhance the tick's attachment behavior. This study evaluated the histological effects of permethrin exposure on the salivary glands and neuroendocrine organs of unfed, virgin H. dromedarii ticks of uniform age. Three fabric treatments consisting of unwashed-untreated (control), washed after treatment (0.125 mg [AI] / cm2) and unwashed-treated were used after 5- and 10 min exposure times for unfed, unmated females. For all of the organs examined, the cellular structure of treated ticks differed from controls as evidenced by increases in cellular activity, as well as significant increases in the size of the cells of the organs under study (P < 0.05). These data conclusively demonstrate that an unexpected enhanced attachment response observed in this tick species after permethrin exposure is the direct result of increased neurosecretory and salivary gland activity induced by that exposure. PMID- 15535584 TI - Cyt1A from Bacillus thuringiensis restores toxicity of Bacillus sphaericus against resistant Culex quinquefasciatus (Diptera: Culicidae). AB - The 2362 strain of Bacillus sphaericus, which produces a binary toxin highly active against Culex mosquitoes, has been developed recently as a commercial larvicide. It is being used currently in operational mosquito control programs in several countries including Brazil, France, India, and the United States. Laboratory studies have shown that mosquitoes can develop resistance to B. sphaericus, and low levels of resistance have already been reported in field populations in Brazil, France, and India. To develop tools for resistance management, the Cyt1A protein of Bacillus thuringiensis subsp. israelensis De Barjac was evaluated for its ability to suppress resistance to B. sphaericus in a highly resistant population of Culex quinquefasciatus Say. A combination of B. sphaericus 2362 in a 10:1 ratio with a strain of B. thuringiensis subsp. israelensis that only produces Cyt1A reduced resistance by >30,000-fold. Resistance was suppressed completely when B. sphaericus was combined with purified Cyt1A crystals in a 10:1 ratio. Synergism was observed between the Cyt1A toxin and B. sphaericus against the resistant mosquito population and accounted for the marked reduction in resistance. However, no synergism was observed between the toxins against a nonresistant mosquito population. These results indicate that Cyt1A could be useful for managing resistance to B. sphaericus 2362 in Culex populations, and also provide additional evidence that Cyt1A may synergize toxicity by enhancing the binding to and insertion of toxins into the mosquito microvillar membrane. PMID- 15535585 TI - Populations of Ixodes scapularis (Acari: Ixodidae) are modulated by drought at a Lyme disease focus in Illinois. AB - From 1990 through 1997, Ixodes scapularis Say larvae and nymphs were sampled between May and October along a 400-m segment of a nature trail in a Lyme disease endemic site in northern Illinois. Ticks were removed from Peromyscus leucopus mice and collected via tick drags at approximately 3-wk intervals. Mouse population estimates along the trail varied from 2, in the spring of 1996 following a year of drought, to > 200 in 1993, the wettest year on record. During the 8-yr period, there were major droughts during the summers of 1991 and 1995. Cumulative degree-days were positively correlated with the number of ticks collected on drags in the same year and negatively correlated with larval tick populations for the following year (P < 0.05). Cumulative rainfall was positively correlated with larval tick abundance for the following year. This was most readily apparent by examination of the larval density on captured mice. In the year following each of two drought years, larval densities were significantly depressed compared with the 8-yr average at the site. PMID- 15535586 TI - Horn fly (Diptera: Muscidae) saliva targets thrombin action in hemostasis. AB - The horn fly, Hematobia irritans (L.), is an important pest of livestock because the adult stage of both sexes are aggressive blood-feeders. Remarkably, even though horn fly adults feed recurrently on their hosts as ectoparasites, these flies lack the ADP-responsive antiplatelet aggregation and vasodilatory antihemostatic systems described for other blood-feeding Diptera. Horn fly salivary gland extracts do interfere with the normal coagulation process as demonstrated by the recalcification time assay. Using this as a baseline, the effects of saliva on recalcification time, activated partial thromboplastin time, prothrombin time, and thrombin time were measured to determine which arm(s) of the coagulation cascade might be impacted. Factor-deficient plasma assays also were used to measure possible perturbations in clotting. Gland-free saliva delayed the recalcification time as well as the activated partial thromboplastin time, prothrombin time, and thrombin time. Saliva also further delayed clotting times of plasmas deficient in factor V, factor VIII, and factor XIII, indicating that other factors in the coagulation cascade were inhibited. Although horn fly saliva did not alter the ability of deficient plasma reconstituted with factor X to clot, it did inhibit deficient plasma reconstituted with factor II (thrombin). Antithrombin activity in saliva was confirmed by its ability to interfere with thrombin hydrolysis of fibrinogen, its normal substrate, and by its inhibition of thrombin action on a chromagenic substrate that mimics the hydrolytic site of fibrinogen. Thus, horn fly saliva contains a factor that specifically targets thrombin, a key component in the coagulation cascade. While the biochemical mechanisms of inhibition may vary, this antihemostatic characteristic is shared with other zoophilic Diptera such as black flies, Simulium spp., and tsetse, Glossina morsitans morsitans Westwood, that feed on ungulates. PMID- 15535588 TI - Integration of repellents, attractants, and insecticides in a "push-pull" strategy for managing German cockroach (Dictyoptera: Blattellidae) populations. AB - "Push-pull" is a behavior manipulation strategy in which behavior-modifying stimuli are integrated with a pest control agent. We evaluated the efficacy of an insecticide bait in combination with attractants ("pull"), repellents ("push"), or both ("push-pull") using a hydramethylnon-based bait, feces-contaminated surfaces as an attractant, and methyl neodecanamide-treated surfaces to repel cockroaches. Both adult males and first-instar German cockroaches, Blattella germanica (L.), chose shelters nearest the attractant-treated surfaces and farthest from the repellent-treated surfaces. Food consumption was highest from food nearest the preferred shelters, and mortality was highest when the insecticide bait was near the preferred shelter. These patterns were more apparent in first instars than in adults. Our results from large arena studies in the laboratory show that the push-pull strategy can be used to displace pests from resources or commodities that are to be protected, and simultaneously lure the pest to an attractant source coupled with a pest control agent. Concentrating cockroaches into a limited area should facilitate the precision-targeting of the pest population and promises to reduce insecticide use. PMID- 15535587 TI - Quantities of Yersinia pestis in fleas (Siphonaptera: Pulicidae, Ceratophyllidae, and Hystrichopsyllidae) collected from areas of known or suspected plague activity. AB - We used a quantitative competitive polymerase chain reaction (PCR) (QC-PCR) to determine bacterial loads in 669 fleas collected in areas of confirmed and suspected plague epizootics. Fleas were collected out of rodent burrows (67.9%) and off of captured animals (24.1%) and rodent carcasses (8.1%). An initial PCR screening assay indicated that 12.1% (81/669) of all fleas were positive for Yersinia pestis. Fleas collected from burrows had significantly lower (chi2 = 264.9, P < 0.0001) infection rates (6.8%) but significantly higher (Student t test, P < 0.0001) bacterial loads (mean = 10(5.6) Y. pestis per flea) than fleas collected off of rodent carcasses (infection rate = 92.6%; mean bacterial load = 10(4.8) Y. pestis per flea). None of the fleas collected off of captured animals were positive for Y. pestis by PCR, although seven of the 176 captured animals were serologically positive for Y. pestis. PMID- 15535589 TI - Laboratory transmission of Rift Valley fever virus by Phlebotomus duboscqi, Phlebotomus papatasi, Phlebotomus sergenti, and Sergentomyia schwetzi (Diptera: Psychodidae). AB - We examined the potential for Phlebotomus papatasi (Scopoli), Phlebotomus duboscqi (Neveu-Lemarie), Phlebotomus sergenti (Parrot), and Sergentomyia schwetzi (Adler, Theodor, & Parrot) to transmit Rift Valley fever (RVF) virus. After feeding on hamsters that had been inoculated with RVF virus, P. papatasi, P. sergenti, and S. schwetzi became infected and developed disseminated infections. All P. papatasi and P. duboscqi inoculated with RVF virus developed high-titer infections. In contrast, only 41% of the inoculated S. schwetzi contained detectable virus, and infected individuals contained significantly less virus than the two Phlebotomus species. Although 50% of the inoculated P. duboscqi transmitted RVF virus to hamsters, only 14% of P. papatasi and none of the S. schwetzi transmitted this virus. Additional studies are needed to determine the role of sand flies as vectors of RVF virus. PMID- 15535590 TI - Seasonal distribution and abundance of ticks (Acari: ixodidae) in northwestern Florida. AB - A 2-yr study was conducted in a northwestern Florida state park and recreation area to determine tick species composition, seasonal abundance, and spatial distribution. Risk of tick attachment to park visitors was also assessed relative to tick abundance from several habitats. Tick collections consisted of weekly cloth drag samples obtained from ground and emergent vegetation along a deer trail, a walking trail located in a 3-mo-old control burned area of forest, a nonburned walking trail, picnic area, and campsites. Sampling was conducted from February 1994 through February 1996. Ticks collected during the study were as follows in descending order of abundance: Ixodes scapularis Say, Amblyomma americanum (L.), A. maculatum Koch, and Dermacentor variabilis (Say). Significantly more (<0.05) adult ticks were recovered from vegetation compared with ground samples only for I. scapularis. Adult questing I. scapularis were collected from October through May (peak December). A. americanum adults were collected from March through August (peak May). Adult A. maculatum were collected during August and September and adult D. variabilis were collected July and August. Larval and nymphal stages of questing A. americanum were collected from June through November (peak July) and February through October (peak September), respectively. A. maculatum nymphs were collected from February through April, June, and September (peak March). No nymphs of I. scapularis were collected. Only four larvae of D. variabilis were collected during the study, all during February 1995. No I. scapularis or A. maculatum larvae were collected from vegetation or ground drags. The greatest risk for tick attachment was in campsites where approximately 60% of all adult and nymphal host-seeking ticks were collected. The next greatest risk area was the walking trail located in the control burned area where an additional 28% of ticks were obtained. PMID- 15535591 TI - Ectoparasites of neonate Indiana bats, Myotis sodalis (Chiroptera: Vespertilionidae), with description of male of Paraspinturnix globosa (Acari: Spinturnicidae). AB - A total of 145 ectoparasites was found on five pups of the Indiana bat, Myotis sodalis, from a roost in Warren County, in southwestern Ohio. Most were Macronyssus crosbyi, the common mite of this host, but three males, two nymphs and a nonengorged female of Paraspinturnix globosa were found. Males and nymphs of this species have not previously been reported. In addition, two female adults of Paraspinturnix globosa were found in the anus of one adult female. The male, female deutonymph and nongravid engorged female of P. globosa are described for the first time. PMID- 15535592 TI - Overwintering and establishment of Aedes albopictus (Diptera: Culicidae) in an urban La Crosse virus enzootic site in Illinois. AB - In 1997, Aedes albopictus (Skuse) was discovered in Peoria, IL, a known focus of La Crosse (LAC) virus transmission. This accidental introduction provided an opportunity to determine whether Ae. albopictus would reemerge in the spring or summer and, if successful overwintering occurred, to follow changes in the geographic range of Ae. albopictus, and to compare its distribution to that of the local treehole mosquito and LAC vector Aedes triseriatus (Say). In 1998, 25 oviposition traps were placed in and around the area of the initial finding of Ae. albopictus, with adult collections by aspirators and larval collections from water-holding containers used to identify areas of additional activity. Ae. albopictus successfully survived the mild 1997-1998 El Nino winter, and expanded its range during 1998. By September 1998, Ae. albopictus oviposited in all 25 traps, including traps near and in wooded sites. Intensity of oviposition activity (number of eggs per positive trap) ranged from 20-40, lower than the range for Ae. triseriatus, which was as high as 175 eggs per infested trap in mid August. Prevalence of Ae. albopictus increased through September, where as the prevalence of Ae. triseriatus declined starting in mid-July. Although direct competition between the two mosquito species cannot be inferred based on this surveillance effort, a gradual range expansion was observed, and Ae. albopictus eggs were collected in traps where initially only Ae. triseriatus oviposition activity was detected. PMID- 15535593 TI - Ovipositional behavior of the horn fly (Diptera: Muscidae) in the field. AB - In the field, the female horn fly obtained blood meals from the bovine host before oviposition. The female flies moved to the lower portion of the hind legs as the legs were spread, and the tail was raised before excretion. Females deposited their eggs on the manure after excretion. The fly flew onto the manure pat for oviposition as the cow walked forward after excretion. It appeared that flies located on the belly received some stimuli for oviposition from the cow just before excretion. The horn fly deposited most of its eggs during the day with occasional deposition at night. Egg deposition increased at 10 degrees C. Fewer flies were observed on manure pats that contained >90% or <84% water. The location of horn flies on the cow and environmental factors associated with oviposition in the field are discussed. PMID- 15535594 TI - Experimental detection of Rift Valley fever virus by reverse transcription polymerase chain reaction assay in large samples of mosquitoes. AB - A reverse transcription-polymerase chain reaction (RT-PCR) was assessed in laboratory tests to detect the presence of single Aedes aegypti (L.) or Eretmapodites quinquevittatus Theobald mosquitoes infected with Rift Valley fever virus in pools of mosquitoes, 50-600 in size, from laboratory colonies or mixed field collections. The viral RNA was detected in all pools containing infected mosquitoes and was shown to be as sensitive as infant mice but more sensitive than Vero cell cultures for virus detection. Pools diluted down to the equivalent of 1:16 000 mosquitoes were also positive by RT-PCR. RNAs from 4 other phleboviruses were negative, there were no false positives and the procedure followed, with the 2 particular primers chosen, gave consistently clear bands of the PCR products on agarose gels without nested PCR being necessary. PMID- 15535595 TI - Responses of adult Ixodes scapularis (Acari: Ixodidae) to urine produced by white tailed deer of various reproductive conditions. AB - The responses of adult female blacklegged ticks, Ixodes scapularis Say, to urine from white-tailed deer, Odocoileus virginianus (Zimmermann), belonging to 4 reproductive categories (doe in estrous, doe out of season, reproductive [dominant] buck, young buck) and to a mixture of urine from nondominant bucks in rut, young bucks out of rut, and nonestrous does were studied in laboratory behavioral bioassays. In high humidity (approximately 95% RH) in a glove box there were no statistically significant arrestment responses to any of the 5 types of urine, but an avoidance response was observed to urine from dominant reproductive bucks. When ticks were tested at approximately 50% RH, with samples of all 5 types of urine in the glove box, significant arrestant responses by the ticks were elicited by urine from does in estrous and by dominant reproductive bucks. When tested without other types of urine in the glove box, the urine mixture elicited an arrestant response at 50% RH. In some circumstances, adult I. scapularis may possibly use deer urine as a chemical cue in selecting host-ambush sites. PMID- 15535596 TI - Evidence of the specific status of Anopheles flavirostris (Diptera: Culicidae). AB - Anophelesflavirostris (Ludlow) from Lombok Island, Indonesia, was crossed with An. minimus species A (CM strain) from Thailand and species E (ISG strain), a new sibling species, from Japan, to determine genetic compatibility. We also compare the scanning micrographs of female cibarial armature of these three species. Both An. minimus CM and ISG females crossed with An. flavirostris males produced eggs without embryos. One An. flavirostris female crossed with an An. minimus CM male deposited unhatchable eggs with an 18% embryonation rate. The scanning micrographs of the cibarial armature clearly show significant differences among An. flavirostris and the two sibling species of the An. minimus complex. These results support the specific status of An. flavirostris. Previous records of An. minimus Theobald in this country are also discussed. PMID- 15535597 TI - Limited transmission of turkey coronavirus in young turkeys by adult Alphitobius diaperinus (Coleoptera: Tenebrionidae). AB - We examined the role of lesser mealworm, Alphitobius diaperinus (Panzer), in the transmission of an enteric disease of turkeys caused by a coronavirus. Turkey coronavirus (TCV) from two sources was studied, one isolate (NC95) was embryo propagated, the second was TCV infected material from turkeys diagnosed with poult enteritis mortality syndrome (PEMS). Beetles were fed virus-infected feces mixed with chicken feed. Transmission of virus was effectively halted by surface sterilization of the beetles. Turkey poults administered beetle homogenates infected with TCV+ PEMS that had not been surface sterilized had reduced weight gains and 50% mortality. Mortality and weight gains were not effected in the NC95 group. Virus isolation procedures were performed to determine NC95 viability at varying time intervals. Beetles were dissected and the guts removed 1, 12, and 24 h after the initial viral feeding. Whole beetles were also examined for comparison. Whole beetles and beetle guts were homogenized and injected into turkey eggs for embryo propagation. Direct immunofluorescence was used to determine the presence of TCV. A. diaperinus were capable of mechanical transmission of TCV. However, only turkey embryos receiving whole beetle and beetle gut homogenates within 1 h of feeding on the virus were positive for TCV. Laboratory studies demonstrating PEMS transmission by A. diaperinus are continuing. PMID- 15535598 TI - Efficiency of drag sampling for estimating population sizes of Ixodes pacificus (Acari: Ixodidae) nymphs in leaf litter. AB - Drag sampling is a commonly used method to obtain relative estimates of the density of questing nymphal Ixodes scapularis Say and I pacificus Cooley & Kohls ticks, which are primary vectors of Lyme disease spirochetes to humans in North America. However, the efficiency of drag sampling in determining absolute population densities of questing nymphs has not been evaluated previously. Therefore, we assessed the efficiency of a single drag-sampling occasion to estimate the total population size of questing I pacificus nymphs in a leaf litter habitat in California. Repeated daily removal sampling was carried out in four areas, each covering 300 m2, on 17 occasions over a 23-d period in the spring of 1999. In total, 573 I. pacificus nymphs were collected, of which 55 (9.6%) were collected on the initial sampling occasion and 20 (3.5%) on the last occasion. The total population size of questing nymphs, i.e., the intersection with the horizontal axis of a linear regression of daily nymphal catch rates on the number of nymphs caught previously, was estimated to be 936. Thus, the efficiency of the initial sampling occasion to estimate the total population size was 5.9% (4.8, 5.0, 5.8, and 9.1%, respectively, for the four individual sampling areas). Further, the overall mean efficiencies of the two, five, and 10 first removal sampling occasions to estimate the absolute nymphal density was 5.2, 4.7, and 4.3%, respectively, and 13 sampling occasions were required to collect 50% of the estimated total nymphal population. PMID- 15535599 TI - [CME radiology 2. Solution. Small cell bronchial carcinoma, adenoid type with infiltrative growth]. PMID- 15535600 TI - [Management of COPD: a review]. AB - COPD is an increasing health problem and one of the leading causes of morbidity and mortality worldwide, associated with immense economic costs. With the recent understanding of the pathogenesis of COPD, primary preventions should provide the best hope to control the rise of this disease and smoking cessation remains the key area to address. The early use of spirometry for diagnosis is of importance, many patients are underdiagnosed until the disease is far advanced. For patients who have developed COPD, a variety of treatment options are available and much can be done to improve the well being of patients with COPD. A review of the management of COPD is presented. PMID- 15535601 TI - [Acute thromboembolism]. AB - The diagnosis of acute pulmonary embolism is based on the assessment of risk factors, consistent symptoms and signs, and the lack of an alternative clinical explanation for the condition of the patient. If the probability of pulmonary embolism is low or intermediate, the diagnosis can be reliably excluded by a negative D-dimer test. A positive D-dimer test requires further evaluation by spiral angio computed tomography technique to confirm or rule out pulmonary embolism. The angiocoexisting pulmonary diseases. If the clinical probability for pulmonary embolism is high an angio-CT rather then a D-dimer should be performed at the first place. If there are signs of deep leg vein thrombosis, massive pulmonary embolism with hemodynamic instability and in other special circumstances the diagnostic approach has to be adapted accordingly. A timely diagnosis and treatment with heparin and oral anticoagulation significantly reduces the acute and chronic morbidity and mortality of pulmonary embolism. PMID- 15535602 TI - [Immunophaenotyping in the diagnosis of lymphoma]. AB - The detection of specific disease markers by immunologic methods is becoming more and more important in lymphoma diagnosis. This has led to the incorporation of immunophaenotypic findings into the new WHO-classification. In comparison with conventional immunohistochemistry, flow cytometry bears the advantage of simultaneous demonstration of several antigens on the same cell. The information obtained allows reliable lineage determination, often demonstration of clonality and it helps in classifying lymphoproliferative diseases. It must be stressed, that also with the new classification, correct diagnoses can only be obtained by a synthesis of morphologic, immunophaenotypic and in certain cases cytogenetic findings. PMID- 15535603 TI - [A systematic approach to the bleeding patient]. AB - As there is no single, simple and reliable laboratory method to test the entire hemostasis the evaluation of patients with hemorrhagic complications remains a multistep process. A detailed history and clinical examination is central in the planning and interpretation of laboratory tests. The present work emphasizes on history taking and guides general practitioners in choosing and interpreting appropriate screening tests. PMID- 15535604 TI - [Paul Ehrlich as clinical pathologist]. PMID- 15535605 TI - [Seasick]. PMID- 15535606 TI - Commentary to, "Phlebotomine sand flies (Diptera: Psychodidae) of the Palestinian West Bank: potential vectors of leishmaniasis," by Samir S. Sawalha, Muhamad S. Shtayeh, Haroun M. Khanfar, Alon Warburg, and Ziad A. Abdeen, J. Med. Entomol. 40: 321-328. PMID- 15535607 TI - The macrocyclic lactone "spinosad," a promising insecticide for tsetse fly control. AB - The susceptibility of tsetse flies (Diptera: Glossinidae), Glossina palpalis gambiensis (Vanderplank), and G. m. morsitans (Westwood) to topically applied spinosad, a mixture of insecticidal molecules from the actinomycete Saccharopolyspora spinosa, is almost as high as to deltamethrin. However, susceptibility to spinosad does not differ significantly between teneral and gravid flies, contrary to deltamethrin. Spinosad might be a promising candidate for future tsetse control by the sequential aerial technique. PMID- 15535608 TI - Patterns of exochorion ornaments on eggs of seven South American species of Lutzomyia sand flies (Diptera: Psychodidae). AB - The patterns of exochorion ornaments on eggs of seven South American Lutzomyia sand fly species were analyzed by scanning electron microscopy (SEM): Lutzomyia (Lutzomyia) cruzi (Mangabeira 1938), Lutzomyia (Micropygomyia) evandroi (Costa Lima and Antunes 1936), L. (Nyssomyia) intermedia (Lutz and Neiva 1912), L. longipalpis (Lutz and Neiva 1912), L. migonei (Franca 1920), L. (Nyssomyia) neivai (Pinto 1926), and L. renei (Martins, Falcao, and Silva 1957). Different patterns were observed, which showed the distinction between some species. Egg ornaments in L. cruzi and L. longipalpis appear as single, parallel, unconnected ridges, whereas eggs of L. migonei appear as single, parallel, connected ridges. Eggs of L. (Nyssomyia) intermedia and L. (N.) neivai present a new variation of the single, unconnected, parallel ridges pattern: small tubercles are present, distributed between the ridges. Eggs of L. renei present an elliptical pattern, with most structures connected by straight ridges. Eggs of L. (M.) evandroi present a polygonal pattern, with alternate rows of small and large hexagons. Our data emphasize the advantages of the SEM approach in the study of the exochorion patterns of Lutzomyia eggs and in the distinction of the sand fly species. PMID- 15535609 TI - Description of a cryptic species, Spinturnix bechsteini n. sp. (Acari, Mesostigmata, Spinturnicidae), parasite of Myotis bechsteinii (Kuhl, 1817) (Chiroptera, Vespertilionidae) by using ecoethology of host bats and statistical methods. AB - Using ecoethology of bats and statistical methods, we anticipate the evidence of specific species of spinturnicids associated with Myotis myotis and Myotis bechsteinii (Chiroptera, Vespertilionidae) mixed colonies are unknown. Traditional description of this new crpytic species Spinturnix bechsteini, parasite of M. bechsteinii, is done. Statistical approach is based on analysis of variance and principal component analysis. The current knowledge on parasites of the genus Myotis in the West Palearctic subregion is presented. PMID- 15535610 TI - Taxonomy and distribution of the series pia of the Lutzomyia verrucarum group (Diptera: Psychodidae), with a description of Lutzomyia emberai n. sp. AB - A new species of phlebotomine sand fly, Lutzomyia emberai n. sp, is described and illustrated from specimens collected in a dwelling of the Embera Indian community, situated in the foothills of the Serrania del Baudol on the Colombian Pacific coast. The morphological characteristics of L. emberai n. sp. suggest that it belongs to the series pia of the group verrucarum, easily differentiated from the other members of this group by diagnostic characters on the palps, labro pharynx, thorax, and spermathecal ducts. The discovery of this new phlebotomine raises to seven the number of species in the series pia, including Lutzomyia pia (Fairchild & Hertig, 1961); Lutzomyia reclusa Fernandez & Rogers, 1991; Lutzomyia suapiensis Le Pont, Torrez-Espejo & Dujardin, 1997; Lutzomyia tihuiliensis Le Pont, Torrez-Espejo & Dujardin, 1997; Lutzomyia tocaniensis Le Pont, Torrez Espejo & Dujardin, 1997; Lutzomyia limafalcaoae (Wolff & Galati, 2002); and Lutzomyia emberai Bejarano, Duque & Velez, 2004, n. sp. The taxonomy, distribution, and medical importance of this series are reviewed. PMID- 15535611 TI - A dispersal model for the range expansion of blacklegged tick (Acari: Ixodidae). AB - The blacklegged tick, Ixodes scapularis Say, a vector for the agents of Lyme borreliosis and other diseases, has expanded its range dramatically over the past 20 yr. However, the relative contributions of different vertebrate host species to this expansion have remained largely unexplored. To address this issue, we simulated the expansion of a theoretical tick population across a simple landscape by using a deterministic, spatially explicit, cellular automata model. The model incorporates the ecology of ticks and three vertebrate hosts: white tailed deer, Odocoileus virginianus Zimmermann; white-footed mouse, Peromyscus leucopus Rafinesque; and American robin, Turdus migratorius L. Host contribution to tick dispersal is modeled as a function of tick burden, home range size, and population density. These parameters were determined using published and unpublished data. Our results suggest that 1) hosts with high tick burdens and large home ranges (e.g., deer) play a critical role in I. scapularis range expansion; 2) hosts with small home ranges (e.g., mice) can limit range expansion if they divert a sufficient number of ticks from feeding on more mobile hosts; and (3) birds that migrate annually (e.g., robins) can play a crucial role in tick range expansion. PMID- 15535612 TI - Diel feeding periodicity of larval anopheline mosquitoes on microorganisms and microinvertebrates: a spatial and temporal comparison of Anopheles quadrimaculatus (Diptera: Culicidae) diets in a Michigan pond. AB - Diel feeding activity of third and fourth instars of Anopheles quadrimaculatus Say were studied in a Michigan permanent pond. This field study examined the consumption of microbial and microinvertebrate food resources over a diel (24-h) period between two habitats (open water and vegetated areas). A fluorochromatic stain (4,6-diamidino-2-phenylindole) was used to quantify microbial dietary components within larval guts and habitats. Microbial analyses show that bacteria were the most abundant food type, followed by detritus, algae, and invertebrate parts/protozoans (IPP). Larval consumption of cladocerans was significantly greater at midnight than noon. Larval gut analyses examined every hour over a 24 h period provided significant evidence as to the approximate time larvae switch microinvertebrate dietary resources. Habitat had a significant effect on microinvertebrate consumption by An. quadrimaculatus larvae. Larvae consumed more water mites and rotifers in the open water areas than vegetated zones. We found that An. quadrimaculatus larvae do not preferentially feed on microinvertebrates over a diel period, however, larvae may feed selectively on rotifers in open water habitats. Knowledge of the "feeding area" or microhabitats, with respect to where and when larvae optimally forage as well as particle sizes and food types consumed in the natural habitats will enhance the success of bacteria and other particulate larvicides. PMID- 15535613 TI - Lack of spatial autocorrelation in fine-scale distributions of Ixodes scapularis (Acari: Ixodidae). AB - Spatial patterns of Ixodes scapularis Say, the vector of the Lyme disease agent, have been examined at various geographic scales, demonstrating that distributions of these ticks are spatially autocorrelated at both national and state scales. We tested the hypothesis that distributions of nymphal I scapularis ticks at the fine scale of an endemic community also are spatially autocorrelated. Nymphal tick densities were determined by collecting ticks from 51 and 47 wooded residential properties in a southern Rhode Island town in 2002 and 2003, respectively. The average tick density at residences during 2002 was 51.17 ( +/- 46.04) nymphs per hour, with a range of 3-297 and median of 40.82. In 2003, the average tick density was 44.48 (+/- 38.31) nymphs per hour, with a range of 3-153 and median of 36. Semivariance analysis revealed no spatial autocorrelation in tick densities between residences, likely due to the high variability of tick distributions at this scale. Further analysis of drag-sampling data at individual residences by using Lloyd's patchiness index (m*/m) demonstrated a patchy distribution of nymphs. High variability of nymphal I. scapularis densities may greatly affect predicting spatial patterns of ticks at a fine scale. PMID- 15535614 TI - Influence of heat and vibration on the movement of the northern fowl mite (Acari: Macronyssidae). AB - Heat and vibration are common host-generated cues that ectoparasites use to orient to hosts. Three experiments evaluated effects of heat and vibration on the movement of northern fowl mite, Ornithonyssus sylviarum (Canestrini & Fanzago). Individual arrested mites in an isolation chamber always initiated movement (walking) after substrate vibration (7.8-min walking duration), but only initiated movement 50% of the time (2.8-min walking duration) upon exposure to a 3 degrees C heat fluctuation. Heat fluctuation in combination with vibration extended the period of activity by approximately 50% (11.6-min walking duration) compared with activity initiated by vibration alone. Mites with longer time off host moved for shorter durations. In a choice test, individual mites consistently moved closer to a 35 degrees C heat source 1 or 6 mm away, but not to a heat source 11 mm away. In a circular arena, mites were able to orient accurately to a 35 degrees C heat source and reached the arena edge almost 4 times faster (11.2 s) than mites without a heat source (41.2 s). These results suggest that northern fowl mite is capable of directed thermo-orientation, as well as modulation of activity depending on the type of sensory information perceived. The adaptive significance of this orientation for a "permanent" ectoparasite is discussed. PMID- 15535615 TI - Photoelectric sensing device for recording mosquito host-seeking behavior in the laboratory. AB - A new automatic device for recording the host-seeking behavior of mosquitoes was designed using photoelectric sensors. The host-seeking rhythms of several species of mosquitoes were recorded under laboratory conditions. Use of CO2, in addition to heating and black color to activate mosquito flight, proved to be important for evaluation of the present recording device. The diel rhythms of nonblood-fed Aedes albopictus (Skuse), Aedes aegypti (L.), Culex quinquefasciatus Say, Culex tritaeniorhynchus Giles and Anopheles stephensi Liston females recorded by the device corresponded well with known flight and activity rhythms in field for the same mosquito species. This simple automatic recording device provided accurate information on the flight behaviors of colonized and field collected mosquitoes. PMID- 15535616 TI - A preliminary assessment of genetic differentiation of Triatoma dimidiata (Hemiptera: Reduviidae) in Guatemala by random amplification of polymorphic DNA polymerase chain reaction. AB - The population genetics of Triatoma dimidiata (Latreille, 1811) from five different provinces in Guatemala, including three sylvan and three domestic populations, was investigated by random amplification of polymorphic DNA polymerase chain reaction. There is a high degree of genetic variation in all of the T. dimidiata populations as evidenced by high levels of average expected heterozygosity and polymorphism. Domestic populations are more closely related to each other (D = 0.05-0.085, Nei's genetic distance) than are the sylvan (D = 0.121-0.189). Within the limited sample size of three populations, there was a correlation with geographic and genetic distance for the domestic populations, but not for the sylvan. Surprisingly, one of the sylvan populations was genetically very similar to the domestic populations. The FST demonstrated a high degree of differentiation at the country-wide level (FST = 0.175) and a moderate degree of differentiation within the sylvan (FST = 0.135) or domestic (FST = 0.097) populations. Although these results demonstrated that gene flow is limited between different provinces in Guatemala, hierarchical analysis showed that barriers between the Atlantic and Pacific drainage slopes were not biologically significant limiters of gene flow. PMID- 15535617 TI - Analysis of reproductive isolation between sibling species anopheles Albitarsis sensu stricto and Anopheles deaneorum, two malaria vectors belonging to the Albitarsis complex (Diptera: Culicidae). AB - Complexes of sibling species are common among mosquitoes, and their existence within vector species can have important epidemiological consequences. Anopheles albitarsis sensu stricto and Anopheles deaneorum Rosa-Freitas are two putative vectors of malaria parasites belonging to the Albitarsis species complex (Diptera: Culicidae). Using an induced mating technique, we studied the reproductive isolation between these two closely related species and their reciprocal hybrids. Evidence for hybrid male sterility consistent with Haldane's rule was found. The results indicate that male hybrids show very low insemination rates, probably due to abnormalities in their reproductive organs. In addition, the data show that hybrid males carrying an X chromosome derived from An. deaneorum perform significantly worse than hybrid males carrying an An. albitarsis s.s. X chromosome. PMID- 15535618 TI - Characterization of Wolbachia infections and interspecific crosses of Aedes (Stegomyia) polynesiensis and Ae. (Stegomyia) riversi (Diptera: Culicidae). AB - Prior studies have identified a complicated pattern of interspecific hybridization between members of the Aedes (Stegomyia) scutellaris (Walker) mosquito group, which includes medically important vectors of bancroftian filariasis and dengue. Here, we report that two members of the group, Aedes polynesiensis Marks and Aedes riversi Bohart & Ingram, are both infected with intracellular Wolbachia bacteria. Sequencing of the Wolbachia wsp gene demonstrates that the infections differ from each other and from Wolbachia infections previously reported in mosquitoes. Aedes polynesiensis is the first mosquito identified with a wMel Wolbachia type. Intraspecific crosses of infected and aposymbiotic lines generated via antibiotic treatment show that the Wolbachia infections in both species cause high levels of cytoplasmic incompatibility. Interspecific crosses show that the two species are reproductively isolated. However, repeating the interspecific crosses with aposymbiotic mosquito strains demonstrates that the Wolbachia infections play a role in preventing hybrid offspring. We discuss Wolbachia infections in relation to better defining the evolutionary relationships and causes of speciation within the group, understanding the basis for the observed east-to-west gradient in filarial refractoriness, and developing novel genetic control measures. PMID- 15535619 TI - High malaria transmission intensity due to Anopheles funestus (Diptera: Culicidae) in a village of savannah-forest transition area in Cameroon. AB - An entomological survey was conducted on vectors of malaria in a village of the forest-savannah transition area in Cameroon from February 1999 to October 2000. A total of 2,050 anopheline mosquitoes belonging to eight species were caught 1) after landing on human volunteers, 2) by using pyrethrum spray collections in human dwellings, and 3) in resting sites outdoors. Anopheles funestus Giles was the most abundant species (accounting for 91% of anophelines caught) followed by Anopheles gambiae Giles (7%). Applying polymerase chain reaction led to the identification of all specimens of the An. funestus group as An. funestus sensu stricto and mosquitoes from the An. gambiae complex were mostly An. gambiae sensu stricto of the S molecular form. Malaria transmission was perennial with an entomological inoculation rate estimated at 172 infective bites per person during the period of study. An. funestus was responsible for 88% of the total malaria transmission, with a Plasmodium falciparum circumsporozoite rate of 6.8% and an anthropophilic rate of 99.3%. These results confirm that in high agricultural activity areas, An. funestus can be, by far, the major malaria vector. PMID- 15535621 TI - Efficacy of permethrin-treated uniforms in combination with DEET topical repellent for protection of French military troops in Cote d'Ivoire. AB - In 2000, 22,000 French military personnel were deployed overseas. The French military health service implemented a vector control strategy including personal protection by the use of permethrin preimpregnated battlefield uniforms (BFUs) and the application on the skin of a topical repellent (50% DEET). In 2000, French forces used an industrial process to impregnate cloth with permethrin by soaking it before cut-out of the BFU. A study was implemented in four experimental huts in Cote d'Ivoire to assess the field efficacy of the impregnated BFUs and their resistance to washing. Taking into account the systematic variations in each variable in the field and using a modeling based on logistic regression and discriminant analysis, this study showed that after 6 h without reapplication, the protective effects of the use of DEET as skin repellent was not significant, perhaps due to the high density of Anopheles mosquitoes during the night catching sessions and an average time of effective repellency of < 2 or 3 h in the field. The analysis also showed that the French process of industrial impregnation of permethrin of the BFU offered in 2000 some protection from mosquito bites but not enough to reduce significantly the incidence of malaria among nonimmune troops. No positive or negative interaction was noted when DEET and the impregnated BFUs were used together. PMID- 15535620 TI - Analysis of the copulatory courtship songs of Lutzomyia longipalpis in six populations from Brazil. AB - The sand fly Lutzomyia longipalpis Lutz & Neiva (Diptera: Psychodidae: Phlebotominae), the main vector of Leishmania infantum in the Americas, is believed to be a species complex, although the status of different Brazilian populations is still somewhat unclear. Preliminary analysis of the acoustic signals that are produced during copulation by L. longipalpis males has suggested the existence of three sibling species in Brazil. In the current report, we analyze in more detail a number of parameters of the copulatory courtship songs of L. longipalpis males from four allopatric populations from different parts of the country (Marajo Island, Natal, Jacobina, and Lapinha Cave) and from two sympatric populations from the locality of Sobral, where two types of males can be differentiated by the number of pale spots (one or two pairs) found on the abdomen. We show that males from the localities of Natal, Marajo, and Sobral (two spot morph) have very similar songs composed of successive bursts, which are modulated in frequency and amplitude. No significant differences were found in the song parameters of these three populations. In contrast, one-spot males from Sobral and males from Jacobina and Lapinha produce songs that are made of pulses but with distinct patterns for each population and significant differences in all song parameters studied. The results suggest that the L. longipalpis complex in Brazil is composed of four sibling species and that the differences in song patterns between the populations are consistent with the level of divergence found in the period gene. PMID- 15535622 TI - Sublethal effects of Metarhizium anisopliae (Deuteromycetes) on engorged larval, nymphal, and adult Ixodes scapularis (Acari: Ixodidae). AB - Ixodes scapularis Say adults, nymphs, and larvae were treated with the entomopathogenic fungus Metarhizium anisopliae in a combination of field and laboratory experiments to assess sublethal effects of the fungus on I. scapularis fecundity and body mass. Postengorgement and egg mass weights were 33 and 50% lower, respectively, in adult females treated with M. ansiopliae in the field before engorging on laboratory rabbits. M. anisopliae did not significantly reduce egg mass weight, conversion efficiency, or oviposition period in I. scapularis females treated with the fungus after engorging on white-tailed deer, although only 33% of treated females oviposited. Engorged nymphs and larvae treated with M. anisopliae converted significantly lower percentages of their engorged weight to their molted adult and nymphal weights. This study indicates that M. anisopliae reduces fitness (fecundity and body mass) in all active I. scapularis stages and indicates that its impact as a biocontrol agent might be higher than that suggested by direct mortality alone. PMID- 15535623 TI - Investigation of negative cross-resistance as a resistance-management tool for insecticide-treated nets. AB - Resistance management for insecticide-treated nets (ITNs) remains a challenge. Options are limited, because a safe and highly active insecticide with a persistence of several months is required. These criteria have only been met by pyrethroids, although organophosphates (OPs) and carbamates have been considered as alternatives for impregnation of eave curtains. It has been observed that some pyrethroid-resistant mosquito strains show increased OP susceptibility over pyrethroid-susceptible strains (i.e., negative cross-resistance). The current study investigated whether this phenomenon applies to a range of mosquito species and strains, because a mixture or rotation strategy for resistance management could then be envisaged. Adult female mosquitoes from laboratory strains of Anopheles stephensi Liston, Anopheles gambiae Giles, and Culex quinquefasciatus Say were tested in World Health Organization susceptibility test kits. For An. stephensi, the highly pyrethroid-resistant DUB 234 strain showed the same level of resistance to malathion as the pyrethroid-susceptible DUB S. The malathion resistant ST MAL strain was as susceptible to pyrethroids as the insecticide susceptible BEECH. For An. gambiae, the malathion tolerance of the previously pyrethroid-resistant RSP strain was significantly higher than that of the insecticide-susceptible KWA. For Cx. quinquefasciatus, selection of the QUINQ strain with permethrin abolished preexisting resistance to the OP malathion as pyrethroid resistance increased, rendering the strain more susceptible to malathion than PEL SS. Some indication of negative cross-resistance to malathion was found for the permethrin-resistant MUHEZA strain. The occurrence of negative cross-resistance seems dependent on the history of insecticide selection and is not generally applicable. Resistance management for ITNs will need to use mechanisms other than negative cross-resistance to be effective. PMID- 15535624 TI - Synergy between toxins of Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus. AB - Synergistic interactions among the multiple endotoxins of Bacillus thuringiensis subsp. israelensis de Barjac play an important role in its high toxicity to mosquito larvae and the absence of insecticide resistance in populations treated with this bacterium. A lack of toxin complexity and synergism are the apparent causes of resistance to Bacillus sphaericus Neide in particular Culex field populations. To identify endotoxin combinations of the two Bacillus species that might improve insecticidal activity and manage mosquito resistance to B. sphaericus, we tested their toxins alone and in combination. Most combinations of B. sphaericus and B. t. subsp. israelensis toxins were synergistic and enhanced toxicity relative to B. sphaericus, particularly against Culex quinquefasciatus Say larvae resistant to B. sphaericus and Aedes aegypti (L.), a species poorly susceptible to B. sphaericus. Toxicity also improved against susceptible Cx. quinquefasciatus. For example, when the CytlAa toxin from B. t. subsp. israelensis was added to Bin and Cry toxins, or when native B. t. subsp. israelensis was combined with B. sphaericus, synergism values as high as 883-fold were observed and combinations were 4-59,000-fold more active than B. sphaericus. These data, and previous studies using cytolytic toxins, validate proposed strategies for improving bacterial larvicides by combining B. sphaericus with B. t. subsp. israelensis or by engineering recombinant bacteria that express endotoxins from both strains. These combinations increase both endotoxin complexity and synergistic interactions and thereby enhance activity and help avoid insecticide resistance. PMID- 15535625 TI - Depletion rate of doramectin from blood serum of penned female white-tailed deer (Artiodactyla: Cervidae). AB - Female white-tailed deer, Odocoileus virginianus (Zimmerman), were held in small pens and administered doramectin by free choice of doramectin-coated whole kernel corn, Zea mays L., fed ad libitum with 19% protein deer pellets also being fed ad libitum in a separate container. The mean concentration of doramectin in the serum during treatment was 72.8 ppb. The mean doramectin concentration in the serum decreased to <2 ppb, the lower limit of detection by high-pressure liquid chromatography, by day 14 after termination of treatment after withdrawal of doramectin-treated corn from the diet. PMID- 15535626 TI - Insecticide resistance in Alabama and Florida mosquito strains of Aedes albopictus. AB - The susceptibility of four strains of Aedes albopictus (Skuse) to permethrin, deltamethrin, resmethrin, chlorpyrifos, malathion, propoxur, fipronil, imidacloprid, spinosad, and Bacillus thuringiensis variety israelensis (Bti) was determined. The HAmAal and MAmAal strains were collected in 2002 and 2003, respectively, from Huntsville and Mobile, AL, and the VBFmAal and SFmAal strains were collected in 1998 from Vero Beach and southern Florida, respectively. The HAmAal strain showed a 22-fold elevated level of resistance to deltamethrin compared with the susceptible Ikaken laboratory strain, whereas the VBFmAal strain showed a six-fold lower sensitivity to deltamethrin compared with Ikaken. However, comparison of resistance ratios for deltamethrin at LC50 and LC90 (21 fold) and the gradual slopes of dose-response curves indicated that the field population of this mosquito strain was heterogenous in response to deltamethrin. All four mosquito strains showed elevated levels of resistance to chlorpyrifos, with resistance ratios from 10 to 33. Nevertheless, except for the relatively low resistance to deltamethrin and chlorpyrifos, all mosquito strains showed a similar susceptibility or lower tolerance to the remaining insecticides tested compared with the susceptible Ikaken strain, even though some, such as permethrin, resmethrin, malathion, and Bti, have been used in the field for a long time, especially in Alabama. These results indicate that the development of resistance to insecticides in Ae. albopictus is slow and conventional insecticides, such as permethrin, resmethrin, malathion, and Bti, and relatively new insecticides, such as fipronil, imidacloprid, and spinosad, may all be valuable for the management of this important mosquito. PMID- 15535627 TI - Gene expression and tissue distribution of the major human allergen Bla g 1 in the German cockroach, Blattella germanica L. (Dictyoptera: Blattellidae). AB - Exposure and sensitization to cockroach allergens is an important risk factor for allergic disease in humans. Despite a recent burgeoning of clinical and socioeconomic studies regarding environmental pervasiveness and human exposure to cockroach allergens, little is known about the basic biology of these proteins. The purpose of this study was to ascertain gene expression patterns and the tissue distribution of Blattella germanica allergen 1 (Bla g 1), a perennial indoor environmental allergen, thought to be involved in digestion in cockroaches. We also investigated the relative potential contribution of different life stages of the German cockroach to environmental Bla g 1. Enzyme linked immunosorbent assay was used to quantify the Bla g 1 contents of feces and various anatomical tissues, and Northern blot analysis was used to elucidate tissue-specific expression of Bla g 1. Results showed that the Bla g 1 protein is most prevalent in the midgut, and the Bla g 1 gene is exclusively expressed by midgut cells. Although Bla g 1 is produced by both sexes and all life stages of the German cockroach, adult females produce and excrete significantly more Bla g 1 in their feces than males or nymphs, even when corrected for body mass or mass of voided feces. Our results show that the concentration of Bla g 1 in feces of adult females is 6- to 7- and 30-fold higher than in adult males and nymphs, respectively, probably because females process more food than other life stages of the German cockroach. PMID- 15535628 TI - Susceptibility of urban and rural populations of Aedes albopictus from Sao Paulo State, Brazil, to infection by dengue-1 and -2 viruses. AB - In Brazil, dengue viruses (DENV) are transmitted by Aedes aegypti (L.). However, the recent introduction of Aedes albopictus (Skuse) has caused concern, because some populations of this species also are capable of DENV transmission. Ae. albopictus from rural and urban localities of Sao Paulo State, Brazil, were artificially exposed to infection with DENV-1 and -2 to evaluate their susceptibility. The rates of infection (6.2 and 5.7% for DENV-1; 5.3 and 12% for DENV-2) and dissemination (22.2 and 40% for DENV-1; 0 and 20% for DENV-2) were low in both urban and rural populations, with no significant differences detected between these mosquito populations. PMID- 15535629 TI - Role of nestling mourning doves and house finches as amplifying hosts of St. Louis encephalitis virus. AB - Nestling mourning doves and house finches produced elevated viremias after inoculation with 2-3 log10 plaque-forming units (PFU) of St Louis encephalitis (SLE) virus and infected 67 and 70% of Culex tarsalis Coquillett that engorged upon them, respectively. Mosquito infection rates as well as the quantity of virus produced after extrinsic incubation increased as a function of the quantity of virus ingested and peaked during days 3-5 postinoculation in mourning doves and days 2-4 in house finches. Only female Cx. tarsalis with body titers > or = 4.6 log10 PFU were capable of transmitting virus. Overall, 38% of females infected by feeding on mourning doves and 22% feeding on house finches were capable of transmission. The quantity of virus expectorated was variable, ranging from 0.8 to 3.4 log10 PFU and was greatest during periods when avian viremias were elevated. Our data indicated that nestling mourning doves and house finches were competent hosts for SLE virus and that the quantity of virus ingested from a viremic avian host varies during the course of the infection and determines transmission rates by the mosquito vector. PMID- 15535630 TI - Experimental transmission of Karshi and Langat (tick-borne encephalitis virus complex) viruses by Ornithodoros ticks (Acari: Argasidae). AB - Selected species of mosquitoes and Ornithodoros ticks were evaluated for their potential to transmit Karshi and Langat (tick-borne encephalitis virus complex) viruses in the laboratory. Although there was no evidence of replication of Karshi virus in either of the two mosquito species tested [Ochlerotatus taeniorhynchus (Wiedemann) or Culex pipiens (L.)], Karshi virus replicated in and was transmitted by all three species of Ornithodoros ticks tested (Ornithodoros parkeri Cooley, Ornithodoros sonrai Sautet & Witkowski, and Ornithodoros tartakovskyi Olenev). When inoculated with Karshi virus, 90% of Ornithodoros ticks (44/49) transmitted this virus by bite to suckling mice, and transmission continued to occur for at least 1 yr, the longest extrinsic incubation tested. After feeding on a suckling mouse with a viremia of approximately 10(5) suckling mouse subcutaneous lethal dose. units of Karshi virus per milliliter of blood, all three species of Ornithodoros tested became infected with and transmitted Karshi virus both trans-stadially and horizontally by bite to suckling mice. In addition, female O. tartakovskyi transmitted Karshi virus vertically to their progeny. In a continuation of a previous study, O. sonrai, orally exposed to Langat virus, were able to transmit this virus after >3 yr, the longest interval tested. Therefore, Ornithodoros spp. should be considered as potential vectors and as possible long-term maintenance hosts for Karshi virus and other members of the tick-borne encephalitis virus complex. PMID- 15535631 TI - Effect of dose on house finch infection with western equine encephalomyelitis and St. Louis encephalitis viruses. AB - House finches, Carpodacus mexicanus, were experimentally infected with high and standard doses of western equine encephalomyelitis virus (WEEV) or St. Louis encephalitis virus (SLEV) to determine whether high doses would produce an elevated viremia response and a high frequency of chronic infections. Finches inoculated with approximately100,000 plaque forming units (PFU) of WEEV or SLEV produced viremia and antibody responses similar to those in finches inoculated with approximately 100 PFU of WEEV or 1000 PFU of SLEV, the approximate quantities of virus expectorated by blood-feeding Culex tarsalis Coquillett. Infected finches were held through winter and then necropsied. Only one finch inoculated with the high dose of SLEV developed a chronic infection. Our data indicated that elevated infectious doses of virus may not increase the viremia level or the frequency of chronic infection in house finches. PMID- 15535632 TI - Western equine encephalomyelitis virus infection affects the life table characteristics of Culex tarsalis (Diptera: Culicidae). AB - The life table attributes of Culex tarsalis Coquillett females infected experimentally by feeding on 4 and 6 log10 plaque-forming units (PFU) of western equine encephalomyelitis virus (WEEV) per milliliter of heparinized chicken blood were compared with an uninfected control group. Females continually were offered 10% sucrose and an oviposition substrate and daily a blood meal through a biomembrane feeder. Mortality (dead females) and fecundity (female eggs per female) were monitored daily until all females died. Overall, 94% of 198 females in the two virus-infected groups were positive for WEEV at death when tested by plaque assay; the average body virus titer at death did not differ between groups. WEEV infection significantly altered the life table characteristics of Cx. tarsalis. Life expectancy at infection in days (ex), reproductive effort in female eggs per female per generation (Ro), and generation time (T) in days for the infected cohorts were significantly lower than for the uninfected controls, whereas the reproductive rate (rc) in female eggs per female per day was higher for infected than uninfected cohorts. In agreement with the WEEV infection data that showed similar body titers, there were few differences between the life table parameters for the 4 and 6 log10 PFU treatment groups. Greatest differences were observed for survivorship between days 17-40 when virus titers in infected dying females were greatest. Our data extend recent studies that indicate mosquito infection with encephalitis viruses has a cost of reduced life expectancy and fitness. PMID- 15535633 TI - Susceptibility of Ochlerotatus taeniorhynchus (Diptera: Culicidae) to infection with epizootic (subtype IC) and enzootic (subtype ID) Venezuelan equine encephalitis viruses: evidence for epizootic strain adaptation. AB - To test the hypothesis that adaptation to epizootic mosquito vectors mediates emergence of Venezuelan equine encephalitis virus (VEEV) from enzootic progenitors, experimental infection studies were conducted to determine the susceptibility of Ochlerotatus taeniorhynchus (Wiedemann) to epizootic and enzootic strains. Artificial blood meals containing epizootic subtype IC strains isolated during the 1962-1964, 1992-1993, and 1995 Venezuelan/Colombian epizootics and closely related Venezuelan enzootic subtype ID strains were used to compare infectivity and transmission potential. Their greater infectivity and replication suggested that adaptation of epizootic strains to Oc. taeniorhynchus may have enhanced epizootic transmission during the 1962-1964 and 1995 IC coastal epizootics. However, strains from the small 1992-1993 Venezuelan outbreak that did not extend to coastal regions do not seem to infect this species better than closely related subtype ID strains. Adaptation of VEEV to epizootic vectors such as Oc. taeniorhynchus mosquitoes may be a determinant of some but not all VEE emergence events and may influence spread into coastal regions. PMID- 15535634 TI - First report on high-degree endophilism in Culex tritaeniorhynchus (Diptera: Culicidae) in an area endemic for Japanese encephalitis. AB - An unusually high occurrence of Culex tritaeniorhynchus Giles resting indoors during daytime was observed in a Japanese encephalitis (JE) endemic area, Bellary district, Karnataka, India. Cx. tritaeniorhynchus, the primary vector of JE in this area, was the dominant species collected, and its abundance showed two peaks corresponding to rice crops in the district. More specimens (n = 20,966) were found resting in indoor habitats than in outdoor vegetation (n = 383), despite the availability of outdoor resting sites. This resting behavior is opposite to the Cx. tritaeniorhynchus behavior reported in the majority of JE-affected areas, where this species was captured in small numbers from indoor resting sites. Increased endophilic resting behavior indicates that indoor residual insecticides may provide an effective control method in this area. PMID- 15535635 TI - Anopheles albitarsis s.l. (Diptera: Culicidae) survivorship and density in a rice irrigation area of the state of Sao Paulo, Brazil. AB - Collections of Anopheles albitarsis Arribalzaga s.l. were conducted from May 1996 to June 2000 to estimate the human biting rate, and mark-release-recapture experiments were carried out to estimate the daily survival probability. The density of An. albitarsis females showed a marked seasonal variation, with peaks as high as 629 individuals per human per night in the wet season. The low daily survival probability of 0.61 is an important factor limiting the vectorial capacity of An. albitarsis s.l. in southeastern Brazil. PMID- 15535637 TI - Tetrathiafulvalenes, oligoacenenes, and their buckminsterfullerene derivatives: the brick and mortar of organic electronics. PMID- 15535638 TI - Organic conductors with unusual band fillings. PMID- 15535639 TI - Charge-transfer and energy-transfer processes in pi-conjugated oligomers and polymers: a molecular picture. PMID- 15535640 TI - Toward systematic understanding of diversity of electronic properties in low dimensional molecular solids. PMID- 15535641 TI - Theoretical framework for quasi-one dimensional systems. PMID- 15535642 TI - New trends in the synthesis of pi-electron donors for molecular conductors and superconductors. PMID- 15535643 TI - Bi-TTF, bis-TTF, and related TTF oligomers. PMID- 15535644 TI - Tetrathiafulvalene cyclophanes and cage molecules. PMID- 15535645 TI - Synthesis strategies and chemistry of nonsymmetrically substituted tetrachalcogenafulvalenes. PMID- 15535646 TI - Highly functionalized tetrathiafulvalenes: riding along the synthetic trail from electrophilic alkynes. PMID- 15535647 TI - Dithiadiazafulvalenes: promising precursors of molecular materials. PMID- 15535648 TI - Conducting organic radical cation salts with organic and organometallic anions. PMID- 15535649 TI - Single-component molecular metals with extended-TTF dithiolate ligands. PMID- 15535650 TI - Organic metals and superconductors based on BETS (BETS = bis(ethylenedithio)tetraselenafulvalene). PMID- 15535651 TI - Bis(ethylenethio)tetrathiafulvalene (BET-TTF) and related dissymmetrical electron donors: from the molecule to functional molecular materials and devices (OFETs). PMID- 15535652 TI - Conducting metal dithiolene complexes: structural and electronic properties. PMID- 15535653 TI - Molecular conductors and superconductors based on trihalides of BEDT-TTF and some of its analogues. PMID- 15535654 TI - Activation of hydrogen- and halogen-bonding interactions in tetrathiafulvalene based crystalline molecular conductors. PMID- 15535655 TI - Magnetic molecular conductors. PMID- 15535657 TI - Conducting and magnetic Langmuir-Blodgett films. PMID- 15535656 TI - Magnetic TTF-based charge-transfer complexes. PMID- 15535658 TI - Phthalocyanines-versatile components of molecular conductors. PMID- 15535659 TI - N,N'-dicyanoquinone diimines (DCNQIs): versatile acceptors for organic conductors. PMID- 15535660 TI - Organic conductors: from charge density wave TTF-TCNQ to superconducting (TMTSF)2PF6. PMID- 15535661 TI - Control of electronic properties of molecular conductors by uniaxial strain. PMID- 15535662 TI - Study of molecular conductors by X-ray diffuse scattering. PMID- 15535663 TI - NMR studies on two-dimensional molecular conductors and superconductors: Mott transition in kappa-(BEDT-TTF)2X. PMID- 15535664 TI - Electron spin resonance: a major probe for molecular conductors. PMID- 15535665 TI - Optical properties of two-dimensional organic conductors: signatures of charge ordering and correlation effects. PMID- 15535666 TI - Muon-spin rotation studies of electronic properties of molecular conductors and superconductors. PMID- 15535667 TI - High magnetic fields: a tool for studying electronic properties of layered organic metals. PMID- 15535668 TI - Catalytic signal amplification using a Heck reaction. An example in the fluorescence sensing of CuII. AB - Catalytic signal enhancement using an organometallic reaction is demonstrated. The reactivity of a Heck cross-coupling reaction that creates a fluorophore is modulated by the addition of a polyazacyclam inhibitor. The inhibitor will complex with Cu(II), which restores the activity of the Pd(II). The addition of Cu(II) therefore leads to the generation of fluorescence, thereby creating a very sensitive assay for Cu(II). The rate of the Heck reaction is followed by monitoring emission as a function of time. The rate is proportional to the Cu(II) concentration and correlates to the affinity of the inhibitor to various metals. This strategy represents a general technique that can be exploited with other catalytic organometallic reactions. PMID- 15535669 TI - Complete thermal-unfolding profiles of oxidized and reduced cytochromes C. AB - The complete thermal-unfolding profiles of both oxidized and reduced forms of cytochrome c551 (PA) from mesophilic Pseudomonas aeruginosa and cytochrome c552 (HT) from thermophilic Hydrogenobacter thermophilus were obtained by the newly developed pressure-proof cell compartment installed in a circular dichroic spectrometer, which facilitates protein thermal-unfolding experiments up to 180 degrees C. The thermodynamic cycle, which relates protein stability and redox function, indicated that the redox potentials of PA and HT in the native state are regulated by the stability of the oxidized proteins rather than by that of the reduced ones. PMID- 15535670 TI - Two-state folding observed in individual protein molecules. AB - The folding dynamics of small proteins are often described in terms of a simple two-state kinetic model. Within this notion, the behavior of individual molecules is expected to be stochastic, with a protein molecule residing in either the unfolded or the folded state for extended periods of time, with intermittent rapid jumps across the free energy barrier. However, a direct observation of this bistable behavior has not been made to date. Rather, previous reports of folding trajectories of individual proteins have shown an unexpected degree of complexity. This raises the question whether the simple kinetic properties derived from classical experiments on large ensembles of molecules are reflected in the folding paths taken by individual proteins. Here we report single-molecule folding/unfolding trajectories observed by fluorescence resonance energy transfer for a protein that meets all criteria of a two state-system. The trajectories, measured on molecules immobilized in lipid vesicles, demonstrate the anticipated bistable behavior, with steplike transitions between folded and unfolded conformations. They further allow us to put an upper bound on the barrier crossing time. PMID- 15535671 TI - Dinitrogen activation by titanium sandwich complexes. AB - The activation of dinitrogen by titanium sandwich complexes of the general form (eta5-C5Me4R)2Ti (R = CHMe2, CMe3, SiMe3) has been systematically investigated. Low-temperature, in situ, solution infrared spectroscopy has allowed detection of monomeric bis-dinitrogen complexes of titanium that are isostructural with more familiar dicarbonyl derivatives. One example, (eta5-C5Me4CHMe2)2Ti(N2)2, has also been characterized by X-ray diffraction and reveals weakly activated dinitrogen ligands. From the solution IR data, the relative azophilicity of the titanium sandwich complexes has been established and increases with smaller cyclopentadienyl substituents. PMID- 15535672 TI - Stretched poly(dimethylsiloxane) gels as NMR alignment media for apolar and weakly polar organic solvents: an ideal tool for measuring RDCs at low molecular concentrations. AB - The measurement of residual dipolar couplings (RDCs), meanwhile a standard method for obtaining structural information in biomolecular NMR, requires partial alignment of the sample. Special demands on alignment media so far limit the applicability of this approach to small molecules in organic solvents. Major limitations are the free scalability of alignment and the suppression of residual signals of the alignment medium to allow effective measurement of low concentration samples. Here, we present stretched poly(dimethylsiloxane) (PDMS) cross-linked by beta-rays as an alignment medium with no visible impurities in 1H NMR spectra but a single signal at approximately 0.1 ppm that can easily be removed by slightly modified water suppression methods. Besides the free scalability, its applicability to the measurement of RDCs in small molecules at low concentration is demonstrated on a approximately 12 mM sample of spiroindene. The induced alignment tensor in this case can be predicted reasonably well by a simplified model on the basis of steric interactions only. PMID- 15535673 TI - Bistability and the phase transition in 1,3,2-dithiazolo[4,5-b]pyrazin-2-yl. AB - The molecular radical 1,3,2-dithiazolo[4,5-b]pyrazin-2-yl (PDTA) exhibits magnetic bistability just above room temperature, undergoing a well-defined hysteretic phase change with TC downward arrow = 297(1) K and TC upward arrow = 343(1) K. The crystal structures of the two phases of PDTA have been determined by single-crystal X-ray diffraction at 323(2) K. LT-PDTA consists of diamagnetic (S = 0) nearly superimposed pi-dimer stacks, while that of HT-PDTA comprises slipped stacks of pi-radicals (S = 1/2). The structural interconversion is suggested to proceed via the cooperative breaking and making of intermolecular S- - -N interactions and an inversion symmetry-preserving "domino cascade" of the pi stacked rings. PMID- 15535674 TI - Not ions alone: barriers to ion permeation in nanopores and channels. AB - A hydrophobic pore of subnanometer dimensions can appear impermeable to an ion even though its radius is still much wider than that of the ion. Pores of molecular dimensions can be found, for instance, in carbon nanotubes, zeolites, or ion channel proteins. We quantify this barrier to ion permeation by calculating the potential of mean force from umbrella-sampled molecular dynamics simulations and compare them to continuum-electrostatic Poisson-Boltzmann calculations. The latter fail to describe the ion barrier because they do not account for the properties of water in the pore. The barrier originates from the energetic cost to desolvate the ion. Even in wide pores, which could accommodate an ion and its hydration shell, a barrier of several kT remains because the liquid water phase is not stable in the hydrophobic pore. Thus, the properties of the solvent play a crucial role in determining permeation properties of ions in confinement at the molecular scale. PMID- 15535675 TI - In situ dual-beam coincidence second harmonic generation as a probe of spatially resolved dynamics at electrochemical interfaces. AB - In situ dual-beam coincidence second harmonic generation was used to monitor spatially resolved dynamics at the Pt(111)/CO-saturated 0.1 M HClO4 interface. The results obtained showed that, for potential steps from 30 to about 870 mV vs RHE, the full electrooxidation of CO as evidenced from SHG occurred at different times for the two areas of the Pt(111) surface probed by the beams. On this basis, the diffusional rates of adsorbed CO cannot be assumed to be generally large enough as to render the entire surface in a homogeneous state, as invoked by the mean field approximation model. PMID- 15535676 TI - OMP decarboxylase: an experimental test of electrostatic destabilization of the enzyme-substrate complex. AB - 6-Methylaminouridine 5'-phosphate (MAUMP) inhibits OMP decarboxylase (Ki = 3 x 10 6 M) maximally at pH values where its amino group is uncharged. Comparison of the chemical shift of free [7-13C]-MAUMP in solutions of varying pH, with that of the enzyme-bound species confirms that this inhibitor is bound with its amino group uncharged. This enzyme's apparent lack of affinity for a cationic substituent, located near the position that would ordinarily be occupied by the scissile carboxylate group of the substrate, does not appear to support the view that the E-S complex is destabilized by electrostatic repulsion in the ground state. PMID- 15535677 TI - Formation of benzynes from 2,6-dihaloaryllithiums: mechanistic basis of the regioselectivity. AB - The key elimination step for the formation of 3-chloro- and 3-fluorobenzyne from 2-chloro-6-fluorophenyllithium displays a pronounced solvent-dependent regioselectivity. 6Li and 13C NMR spectroscopic studies on 2-chloro-6 fluorophenyllithium reveal a single monomeric aryllithium, suggested by DFT computational studies to be a trisolvate. Rate studies indicate that the elimination of LiCl and LiF proceeds via trisolvated and disolvated monomers, respectively. PMID- 15535678 TI - Cation pool-initiated controlled/living polymerization using microsystems. AB - The "cation pool" of an N-acyliminium ion was found to serve as an effective initiator of cationic polymerization of vinyl ethers in a microsystem consisting of two micromixers and a microtube reactor. The polymerization led to very narrow molecular weight distribution (Mw/Mn = 1.14). The molecular weight (Mn) increased linearly with an increase in the amount of the monomer. The carbocationic polymer end was effectively trapped by allyltrimethylsilane. The present observations illustrate the potential of microsystems, in conjunction with the cation pool, to effect polymerization in a highly controlled manner without the deceleration inherent in the dynamic equilibrium between active and dormant species. PMID- 15535679 TI - Nickel(II)-catalyzed highly enantioselective hydrophosphination of methacrylonitrile. AB - The dicationic Ni(II) complex [Ni(Pigiphos)(THF)](ClO4)2, [1](ClO4)2 ((R)-(S) Pigiphos = bis-{(R)-1-[(S)-2 (diphenylphosphino)ferrocenyl]ethyl}cyclohexylphosphine), catalyzes the addition of bulky aliphatic secondary phosphines to methaacrylonitrile. This hydrophosphination reaction reaches TON = 900 and enantioselectivities up to 94%. A catalytic cycle involving 1,4-conjugate addition of R2PH to methacrylonitrile is supported by the isolation and characterization of a catalytically active N coordinated, methacrylonitrile Ni complex. PMID- 15535680 TI - Molecule-bridged mixed-valent intermediates involving the Ru I oxidation state. AB - The diruthenium(II) complexes {(mu-L)[RuCl(Cym)]2}(PF6)n, Cym = p-cymene = 4 isopropyltoluene, L = 2,2'-azobispyridine = abpy and n = 1, or L = 2,5-bis(1 phenyliminoethyl)pyrazine = bpip and n = 2, were synthesized and characterized by NMR (n = 2) or EPR spectroscopy (n = 1). Whereas the (n = 1) species are ligand radical-bridged RuIIRuII complexes, the three-electron reduction under loss of both chloride ions produces the ions {(mu-L)[Ru(Cym)]2}+, which could be identified as RuI(4d7)-containing mixed-valent species (Ru0RuI or RuIRuII) through UV-vis-NIR spectroelectrochemistry (intervalence charge-transfer bands around 1500 nm) and EPR (rhombic g tensor anisotropy). The weak metal-metal interaction of the dsigma electrons from the eg set is responsible for the small electrochemical coupling with comproportionation constants Kc approximately 102. PMID- 15535681 TI - Ordered mesoporous ceramics stable up to 1500 degrees C from diblock copolymer mesophases. AB - In the present study, a poly(isoprene-block-dimethylamino ethyl methacrylate) diblock copolymer (PI-b-PDMAEMA) is used to structure-direct a polysilazane pre ceramic polymer, commercially known as Ceraset. To the polymer was added a 2-fold excess in weight of the silazane oligomer (Ceraset). The resulting composite was cast into films, and after cooperative self-assembly of block copolymer and Ceraset, the structure was permanently set in the hexagonal columnar morphology, as evidenced by small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). Cross-linking of the silazane oligomer was achieved with a radical initiator at 120 degrees C. Upon heating of the composite to 1500 degrees C under nitrogen, the structure is preserved and a mesoporous ceramic material is obtained, as demonstrated by SAXS and TEM. The pores are open and accessible, as evidenced by nitrogen sorption/desorption measurements indicating a surface area of about 51 m2 g-1 and a pore diameter of 13 nm, consistent with TEM analysis. These results suggest that the use of block copolymer mesophases may provide a simple, easily controlled pathway for the preparation of various high-temperature ceramic mesostructures. PMID- 15535682 TI - Acidity of secondary hydroxyls in ATP and adenosine analogues and the question of a 2',3'-hydrogen bond in ribonucleosides. AB - As sugar-modified nucleosides, nucleotides, and oligonucleotides are much used in pharmacology and enzymatic studies, knowledge on the relative pKa values of the secondary hydroxyls can be of great help in design and interpretations. To obtain this, we have determined the acidity constants for ATP, dATP, 2'-F-dATP, 2'-NH2 ATP, and 2'-O-methyl ATP in aqueous solution. The influence of the relative acidities seem to be mainly from inductive effects since a good correlation between pKa values and group electronegativity is found. There is no clear energetic contribution from a much suggested H-bond between the 2'-OH and 3' oxyanion in adenosine. To clarify if this kind of H-bond would make a more prominent energetic contribution in less-polar solvents, we also determined the acidity of secondary hydroxyls in adenosine, 2'-O-methyladenosine, and 3'-O methyladenosine in water, methanol, and DMSO. The relative differences in pKa values were, however, quite similar in all solvents, suggesting that no major energetic contribution is made by an intramolecular H-bond in adenosine. PMID- 15535683 TI - Ethylene tetramerization: a new route to produce 1-octene in exceptionally high selectivities. AB - Linear alpha-olefins, such as 1-hexene and 1-octene, are important comonomers in the production of linear low-density polyethylene (LLDPE). The conventional method of producing 1-hexene and 1-octene is by oligomerization of ethylene, which yields a wide spectrum of linear alpha-olefins (LAOs). While there exists several processes for producing 1-hexene via ethylene trimerization, a similar route for the selective production of 1-octene has so far been elusive. We now, for the first time, report an unprecedented ethylene tetramerization reaction that produces 1-octene in selectivities exceeding 70%, using an aluminoxane activated chromium/((R2)2P)2NR1 catalyst system. PMID- 15535684 TI - Structural models of the bimetallic subunit at the A-cluster of acetyl coenzyme a synthase/CO dehydrogenase: binuclear sulfur-bridged Ni-Cu and Ni-Ni complexes and their reactions with CO. AB - The Ni(II)-dicarboxamido-dithiolato complexes (Et4N)2[Ni(NpPepS)] (1) and (Et4N)2[Ni(PhPepS)] (2) were used as Nid metallosynthons in the construction of higher nuclearity dinuclear Ni-Cu and Ni-Ni species to model the bimetallic Mp Nid site of the A-cluster of acetyl coenzyme A synthase/CO dehydrogenase (ACS/CODH). Reaction of 1 with [Cu(neo)Cl] and [Ni(terpy)Cl2] in MeCN affords the dinuclear complexes (Et4N)[Cu(neo)Ni(NpPepS)] (3) and [Ni(terpy)Ni(NpPepS)] (4), respectively. Reaction of 2 with [Ni(dppe)Cl2] in MeCN yields [Ni(dppe)Ni(PhPepS)] (6). The Ni-Cu complex 3 exhibits no redox chemistry at the Nid site and no reaction with CO. In contrast, the Nip sites in 4 and 6 are readily reduced (characterized by their Ni(I) EPR spectra) and bind CO, exhibiting nuco bands at 2044 and 1997 cm-1, respectively, indicating terminal CO binding. The present Ni-Ni systems replicate the structural and chemical properties of the A-cluster site in ACS/CODH and support the presence of Ni at Mp in the catalytically active enzyme. PMID- 15535685 TI - Design of catalytic carbonylative polymerizations of heterocycles. Synthesis of polyesters and amphiphilic poly(amide-block-ester)s. AB - The combination of CH3C(O)Co(CO)3P(o-tolyl)3/m-methoxypyridine serves as a versatile catalyst for both the carbonylative polymerizations of epoxides and N alkylaziridines under closely resembling mechanistic hypotheses. Diblock poly(beta-alanoid-b-beta-hydroxypropionate)s can be synthesized even when the aziridine and epoxide comonomers are added all together at the beginning of the reaction. PMID- 15535686 TI - Bromoporphyrins as versatile synthons for modular construction of chiral porphyrins: cobalt-catalyzed highly enantioselective and diastereoselective cyclopropanation. AB - 5,10-Bis(2',6'-dibromophenyl)porphyrins bearing various substituents at the 10 and 20 positions were demonstrated to be versatile synthons for modular construction of chiral porphyrins via palladium-catalyzed amidation reactions with chiral amides. The quadruple carbon-nitrogen bond formation reactions were accomplished in high yields with different chiral amide building blocks under mild conditions, forming a family of D2-symmetric chiral porphyrins. Cobalt(II) complexes of these chiral porphyrins were prepared in high yields and shown to be active catalysts for highly enantioselective and diastereoselective cyclopropanation under a practical one-pot protocol (alkenes as limiting reagents and no slow addition of diazo reagents). PMID- 15535687 TI - Total synthesis of kendomycin: a macro-C-glycosidation approach. AB - Kendomycin, also known as (-)-TAN 2162, is a novel polyketide-derived ansamycin isolated from Streptomyces sp., which exhibits potent antagonist and agonist activities at the endothelin and calcitonin receptors, respectively. This bacterial metabolite also possesses a strong antibiotic activity against a range of gram-positive and -negative bacteria and cytostatic effects on the growth of human cancer cell lines. When a novel macroglycosidation reaction is employed as the key step, the first enantioselective total synthesis of kendomycin has been accomplished. A Friedel-Crafts-type ring closure of the acyclic precursor containing tetrahydropyran and benzofuran moieties produces the macrocycle as a single stereoisomer in good yield, thus establishing the aryl C-glycosidic linkage of the ansa core. This reaction requires a phenolic glycosyl acceptor and appears to proceed through a rapid O-glycosidation followed by a slow rearrangement to an aryl C-glycoside. The requisite secomacrocycle is prepared by the Pd(0)-catalyzed B-alkyl Suzuki-Miyaura cross-coupling of two subunits, which in turn can be expeditiously assembled from readily available building blocks in a modular fashion. PMID- 15535688 TI - A trimeric HIV-1 fusion peptide construct which does not self-associate in aqueous solution and which has 15-fold higher membrane fusion rate. AB - A peptide construct (FPtr) was synthesized which mimics the biologically relevant topology of fusion peptide (FP) domains of the trimeric HIV-1 gp41 envelope protein. The FP domains play a critical role in gp41-catalyzed fusion of viral and host cell membranes which is a key step in viral infection. The FPtr construct contains three FP strands chemically bonded at their C-termini through lysine side chains. Analytical ultracentrifugation demonstrated that FPtr does not self-associate in aqueous solution and therefore models the expected FP topology of gp41. Comparative functional fusion assays were carried out using FPtr, FPdm (a cross-linked FP dimer construct), and FPmn (FP monomer). The derived fusion rate constants order ktr > kdm > kmn, and the ratio ktr/kmn has values in the range of 15-40. These results suggest that there is strong correlation of the fusion rate with the biologically relevant trimeric FP topology. PMID- 15535689 TI - Enantioselective formal hydration of alpha,beta-unsaturated imides by Al catalyzed conjugate addition of oxime nucleophiles. AB - The (salen)Al-catalyzed asymmetric conjugate addition of salicylaldoxime to alpha,beta-unsaturated imides is the key step in an efficient and highly enantioselective two-step formal hydration of these electron-deficient olefins. This reaction constitutes the first example of an enantioselective conjugate addition of an oxygen-centered nucleophile to alpha,beta-unsaturated carboxylic acid derivatives. Application of this method to chiral, nonracemic substrates revealed a high level of catalyst-induced diastereoselectivity, underscoring its potential utility for polyketide natural product synthesis. PMID- 15535690 TI - Gas-phase IR spectroscopy of anionic iron carbonyl clusters. AB - The first gas-phase vibrational spectra are presented for several anionic iron carbonyl clusters, ranging in size from Fe(CO)4- to Fe5(CO)14- in the CO stretching region (1600-2100 cm-1). The experimental spectra provide some immediate structural information about the clusters in the form of low-wavenumber (1750-1850 cm-1) bands marking the presence of bridging carbonyl ligands (mu2 COs). Supporting DFT calculations are presented for the smaller clusters (<3 Fe atoms) and give good agreement with the experimental data, allowing structural assignments for these cases. The Fe2(CO)7- spectrum suggests a structure lacking bridging carbonyl ligands, in agreement with the DFT results. For the case of Fe2(CO)8-, there are two possible structures based on the calculations, both with and without bridging carbonyls. The presence of a low-frequency band ( approximately 1770 cm-1) in the experimental spectrum conclusively demonstrates the existence of the bridged form. The ramifications of these data for metal metal bonding in the clusters are also considered. PMID- 15535691 TI - Tuning the DNA reactivity of cis-platinum: conjugation to a mismatch-specific metallointercalator. AB - A novel bimetallic conjugate combining a rhodium intercalator that selectively binds DNA mismatches and a reactive cis-platinum analogue that targets DNA by coordination has been prepared. The site-selectivity of the bimetallic complex in forming coordination adducts is examined using mismatched and well-matched oligonucleotides of different sequences. The results indicate that through the bifunctional complex, the platinum center can be targeted near mismatched sites. Interestingly, with mismatched, DNA both intrastrand and the less common interstrand cross-linked adducts are formed. The recognition of a DNA mismatch by the bulky Rh intercalator appears to direct the Pt unit, depending upon steric contraints, to react preferentially with mismatched DNA at a site that may or may not be the preferred site of Pt coordination. Thus, the presence of a permanent link to a site-specific intercalator is able to tune the reactivity of the cis platinum analogue. PMID- 15535692 TI - Chemoselective attachment of biologically active proteins to surfaces by expressed protein ligation and its application for "protein chip" fabrication. AB - The present work describes a general method for the selective attachment of proteins to solid surfaces through their C-termini that can be used for the efficient creation of protein chips. Our method is based in the chemoselective reaction between a protein C-terminal alpha-thioester and a modified surface containing N-terminal Cys residues. alpha-Thioester proteins can be obtained using standard recombinant techniques by using expression vectors containing engineered inteins. This new method was used to immobilize two fluorescent proteins and a functional SH3 domain using a protein microarrayer. PMID- 15535693 TI - Photostimulated hole transport through a DNA duplex immobilized on a gold electrode. AB - Photostimulated hole transport through DNA duplexes immobilized on gold electrodes has been investigated. By modifying a gold electrode with a DNA duplex containing a photosensitizer, we have observed a sequence-dependent cathodic photocurrent. DNA acts as a good mediator for cathodic photocurrent when appropriate sequences are selected. PMID- 15535694 TI - Effects of H2O and D2O on polyproline II helical structure. AB - The interaction of solvent with a polypeptide chain is one of the primary factors controlling protein folding and stability. In biologically relevant systems, this solvent is most often water. Experimental estimates of the role of water in peptide folding can be obtained from solvent perturbation experiments. The simplest perturbant for H2O water is its isotopic D2O form. The solvation of peptides known to form PII helices with D2O versus H2O increases their propensity to adopt the PII conformation. PMID- 15535695 TI - Enantioselective sensing of chiral carboxylic acids. AB - A chiral 1,8-diacridylnaphthalene-derived fluorosensor has been prepared and used for enantioselective sensing of a broad variety of chiral carboxylic acids including amino acids, aliphatic acids, arylalkanoic acids, and halogenated carboxylic acids. Fluorescence titration experiments in acetonitrile gave linear Stern-Volmer plots for 1:1 and 1:2 complexes and enantioselectivities up to 4.5. PMID- 15535696 TI - Formation of a supramolecular hyperbranched polymer from self-organization of an AB2 monomer containing a crown ether and two paraquat moieties. AB - An AB2 monomer containing a bis(m-phenylene)-32-crown-10 and two paraquat moieties was designed and prepared. It self-organized to form a supramolecular hyperbranched polymer in solution. PMID- 15535697 TI - Relationship of stereochemical and skeletal diversity of small molecules to cellular measurement space. AB - Systematic and quantitative measurements of the roles of stereochemistry and skeleton-dependent conformational restriction were made using multidimensional screening. We first used diversity-oriented synthesis to synthesize the same number (122) of [10.4.0] bicyclic products (B) and their corresponding monocyclic precursors (M). We measured the ability of these compounds to modulate a broad swath of biology using 40 parallel cell-based assays. We analyzed the results using statistical methods that revealed illuminating relationships between stereochemistry, ring number, and assay outcomes. Conformational restriction by ring-closing metathesis increased the specificity of responses among active compounds and was the dominant factor in global activity patterns. Hierarchical clustering also revealed that stereochemistry was a second dominant factor; whereas the stereochemistry of macrocyclic appendages was a determinant for bicyclic compounds, the stereochemistry of the carbohydrates was a determinant for the monocyclic compounds of global activity patterns. These studies illustrate a quantitative method for measuring stereochemical and skeletal diversity of small molecules and their cellular consequences. PMID- 15535698 TI - Probing molecular interfaces using 2D magic-angle-spinning NMR on protein mixtures with different uniform labeling. AB - A general NMR strategy to directly study molecular interfaces under magic angle spinning is introduced. The approach is based on the spectroscopic analysis of uniformly, but heterogeneously, labeled molecular mixtures containing the spin species X and Y (X:Y). For the case of an ((15)N:(13)C) labeled sample, the use of NC, NHC, and NHHC transfers is demonstrated. Applied to the ((13)C:(15)N) labeled dimeric form of the 85 amino acid protein Crh, the NHHC approach reveals a variety of monomer-monomer interactions in the microcrystalline state. PMID- 15535699 TI - Kinetic studies of AMP-dependent phosphorolysis of amylopectin catalyzed by phosphorylase b on a 27 MHz quartz-crystal microbalance. AB - Catalytic cleavage reactions of phosphorylase b were monitored directly on an amylopectin-immobilized 27 MHz quartz-crystal microbalance (QCM). When the inactivated phosphorylase b was injected into a phosphate buffer solution of amylopectin-immobilized QCM (method A), the binding of the enzyme to amylopectin was observed as a frequency decrease (mass increase). Then, when AMP (adenosine monophosphate) was added to activate the enzyme, the frequency gradually increased (mass decreased) due to the phosphorolysis of amylopectin in the presence of phosphates as buffers. When the AMP-activated phosphorylase b was employed (method B), the continuous reaction was observed which includes both the mass increase due to the enzyme binding to amylopectin at first and then the following mass decrease due to the phosphorolysis by the AMP-activated enzyme. All kinetic parameters for the enzyme binding to the substrate (binding and dissociation rate constants, k(on) and k(off), and dissociation constant, K(d)), the AMP binding to the enzyme as activator (K(AMP)), the catalytic rate constant (k(cat)) were obtained from curve fittings of time-courses of frequency (mass) changes. The obtained kinetic parameters were compared with those from Michaelis Menten kinetics. PMID- 15535700 TI - HIV fusion inhibitor peptide T-1249 is able to insert or adsorb to lipidic bilayers. Putative correlation with improved efficiency. AB - T-1249 is a HIV fusion inhibitor peptide under clinical trials. Its interaction with biological membrane models (large unilamellar vesicles) was studied using fluorescence spectroscopy. A gp41 peptide that includes one of the hydrophobic terminals of T-1249 was also studied. Both peptides partition extensively to liquid-crystalline POPC (1-palmitoyl-2-oleyl-sn-glycero-3-phosphocholine) (DeltaG = -7.0 kcal/mol and -8.7 kcal/mol, for T-1249 and terminal peptide, respectively) and are located at the interface of the membrane. T-1249 is essentially in a random coil conformation in this lipidic medium, although a small alpha-helix contribution is present. When other lipid compositions are used (DPPC, POPG + POPC, and POPC + cholesterol) (DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) and POPG (1-palmitoyl-2-oleyl-sn-glycero-3-[phospho-rac-(1-glycerol)), partition decreases, the most severe effect being the presence of cholesterol. Partition experiments and fluorescence resonance energy transfer analysis show that T-1249 adsorbs to cholesterol-rich membranes. The improved clinical efficiency of T-1249 relative to enfuvirtide (T20) may be related to its bigger partition coefficient and ability to adsorb to rigid lipidic areas on the cell surface, where most receptors are inserted. Moreover, adsorption to the sterol-rich viral membrane helps to increase the local concentration of the inhibitor peptide at the fusion site. PMID- 15535701 TI - Dynamics and mechanisms of the multiphoton gated photochromic reaction of diarylethene derivatives. AB - The cycloreversion (ring-opening) process of one of the photochromic diarylethene derivatives, bis(2-methyl-5-phenylthiophen-3-yl)perfluorocyclopentene, was investigated by means of picosecond and femtosecond laser photolysis methods. The drastic enhancement of the reaction yield was observed only by the picosecond laser exposure. The excitation intensity effect of the reaction profiles revealed that the successive multiphoton absorption process leading to higher excited states opened the efficient cycloreversion process with a reaction yield of (50 +/- 10)%, while the one-photon absorption directly pumped to a higher excited state did not lead to the efficient cycloreversion reaction. These results indicate that not the energy of the excitation but the character of the electronic state takes an important role in the enhancement of the cycloreversion reaction. PMID- 15535702 TI - Recognition-induced transformation of microspheres into vesicles: morphology and size control. AB - Polystyrene functionalized with diamidopyridine (DAP) recognition units self assembles in nonpolar media to form thermally reversible micrometer-scale spherical aggregates. The size and the thermal stability of these microspheres can be controlled by the molecular weight of the polymer. The addition of thymine functionalized polymer to these self-assembled microspheres converted them into vesicular aggregates with a controlled size. The morphology change was reversible: the addition of DAP-functionalized polymer converted the vesicles back to microspheres. PMID- 15535703 TI - Crankshaft motion in a highly congested bis(triarylmethyl)peroxide. AB - Crankshaft motion has been proposed in the solid state for molecular fragments consisting of three or more rotors linked by single bonds, whereby the two terminal rotors are static and the internal rotors experience circular motion. Bis-[tri-(3,5-di-tert-butyl)phenylmethyl]-peroxide 2 was tested as a model in search of crankshaft motion at the molecular level. In the case of peroxide 2, the bulky trityl groups may be viewed as the external static rotors, while the two peroxide oxygens can undergo the sought after internal rotation. Evidence for this process in the case of peroxide 2 was obtained from conformational dynamics determined by variable-temperature (13)C and (1)H NMR between 190 and 375 K in toluene-d(8). Detailed spectral assignments for the interpretation of two coalescence processes were based on a correlation between NMR spectra obtained in solution at low temperature, in the solid state by (13)C CPMAS NMR, and by GIAO calculations based on a B3LYP/6-31G structure of 2 obtained from its X-ray coordinates as the input. Evidence supporting crankshaft rotation rather than slippage of the trityl groups was obtained from molecular mechanics calculations. PMID- 15535704 TI - Why are proton transfers at carbon slow? Self-exchange reactions. AB - When the quantum character of proton transfer is taken into account, the intrinsic slowness of self-exchange proton transfer at carbon appears as a result of its nonadiabatic character as opposed to the adiabatic character of proton transfer at oxygen and nitrogen. This difference is caused by the lesser polarity of C-H bonds as compared to that of O-H and N-H bonds. Besides solvent and heavy atom intramolecular reorganizations, the kinetics of the reaction are consequently governed at the level of a pre-exponential term by proton tunneling through the barrier. These contrasting behaviors are illustrated by an analysis of the CH(3)H + (-)CH(3), H(2)O + OH(-), and (+)NH(4) + NH(3) self-exchange reactions. The effect of electron-withdrawing substituents and the case of cation radicals are discussed within the same framework taking the O(2)NCH(2)H + CH(2)=NO(2)(-) and (+.)H(2)NCH(2)H + (.)CH(2)NH(2) as examples. Illustrated by the CH(2)=CH-CH(2)H + (-)CH(2)-CH=CH(2) couple, it is shown that the "imbalanced character of the transition state" is related to heavy-atom intramolecular reorganization. Combination of these various effects is finally analyzed, taking the O(2)N-CH(2)=CH-CH(2)H + CH(2)=CH-CH=NO(2)(-) and (+.)H(2)N-CH(2)=CH-CH(2)H + (.)CH(2)-CH=CH(2)-NH(2) couples as examples. PMID- 15535705 TI - The mechanism of aluminum-catalyzed Meerwein-Schmidt-Ponndorf-Verley reduction of carbonyls to alcohols. AB - The mechanistic details of the Meerwein-Schmidt-Ponndorf-Verley (MSPV) reduction of ketones to the corresponding alcohols were investigated both experimentally and computationally. Density functional theory (DFT) was used to assess the energetics of several proposed pathways (direct hydrogen transfer, hydridic, and radical). Our results demonstrate that a direct hydrogen transfer mechanism involving a concerted six-membered ring transition state is the most favorable pathway for all calculated systems starting from a small model system and concluding with the experimentally investigated BINOLate/Al/(i)PrOH/MePhC=O system. Experimental values for the activation parameters of acetophenone reduction using the BINOLate/Al/(i)PrOH system (DeltaG# = 21.8 kcal/mol, DeltaH# = 18.5 kcal/mol, DeltaS# = -11.7 au) were determined on the basis of kinetic investigation of the reaction and are in good agreement with the computational findings for this system. Calculated and experimental kinetic isotope effects support the concerted mechanism. PMID- 15535706 TI - Synthesis and reactivity of the hydrido- and alkylrhenium methylidene complexes Cp*(PMe3)2(R)Re=CH2 (R = H, CH3). AB - Protonolysis of the dimethylrhenium(III) compound Cp(PMe(3))(2)Re(CH(3))(2) (3) led to formation of the highly reactive hydridorhenium methylidene compound [Cp(PMe(3))(2)Re(CH(2))(H)][OTf] (4), which was characterized spectroscopically at low temperature. Although 4 decomposed above -30 degrees C, reactivity studies performed at low temperature indicated it was in equilibrium with the coordinatively unsaturated methylrhenium complex [Cp(PMe(3))(2)Re(CH(3))][OTf] (2). Methylidene complex 4 was found to react with PMe(3) to afford [Cp(PMe(3))(3)Re(CH(3))][OTf] (6) and with chloride anion to give Cp(PMe(3))(2)Re(Me)Cl (7). When BAr(f) anion was added to 4, the thermally stable methylrhenium methylidene complex [Cp(PMe(3))(2)Re(CH(2))(CH(3))][BAr(f)] (8) was isolated upon warming to room temperature. The mechanisms of formation of both 4 and 8 are discussed in detail, including DFT calculations. The novel carbonyl ligated complex Cp(CO)(2)Re(CH(3))OTf (12) was prepared, isolated, and found to not undergo migration reactions to form methylidene complexes. PMID- 15535707 TI - Perfect orientation ordered in-situ one-dimensional self-assembly of Mn-doped PbSe nanocrystals. AB - We demonstrate a novel approach for the large-scale, shape-controlled synthesis of one-dimensional (1D) corrugated nanoarrays of Pb(1-x)Mn(x)Se nanocrystals (0.002 < or = x < or = 0.008) through an in-situ self-assembly without using either capping polymer or ionic surfactant. The one-step-prepared 1D nanoarrays exhibit a well-defined morphology, single-crystal orientation, and clean surface without amorphous contamination. The average diameter of the 1D nanoarrays can be controlled and varied from <10 to 80 nm by finely tuning the assembly temperature and the growth time. Four growth models were suggested to explain the in-situ self-assembly processes based on the fundamental building blocks of octahedral nanocrystals by sharing [111] facets. PMID- 15535708 TI - Strong electronic coupling between dimolybdenum units linked by the N,N' dimethyloxamidate anion in a molecule having a heteronaphthalene-like structure. AB - Very strong electronic communication, manifested in a K(C) value of ca. 10(9), has been found in a system of three compounds separated by one electron oxidation where each compound contains two metal-metal bonded Mo(2)(n+) units linked by a dimethyloxamidate anion, CH(3)N(O)C-C(O)NCH(3)(2)(-). The Mo-Mo distances increase as the oxidation state increases and the bond order decreases, while the diamagnetic, doubly oxidized species becomes essentially planar and resembles a naphthalene molecule. Calculations at the DFT level indicate that a strong transition in the near-IR region, shown by the singly oxidized and paramagnetic species, is best described as a HOMO-1 --> SOMO transition. PMID- 15535709 TI - "Inverse-electron-demand" ligand substitution: experimental and computational insights into olefin exchange at palladium(0). AB - The mechanism of olefin substitution at palladium(0) has been studied, and the results provide unique insights into the fundamental reactivity of electron-rich late transition metals. A systematic series of bathocuproine-palladium(0) complexes bearing trans-beta-nitrostyrene ligands (ns(X) = X-C(6)H(4)CH=CHNO(2); X = OCH(3), CH(3), H, Br, CF(3)), (bc)Pd(0)ns(X) (3(X)), was prepared and characterized, and olefin-substitution reactions of these complexes were found to proceed by an associative mechanism. In cross-reactions between (bc)Pd(ns(CH)()3) and ns(X) (X = OCH(3), H, Br, CF(3)), more-electron-deficient olefins react more rapidly (relative rate: ns(CF)()3 > ns(Br) > ns(H) > ns(OCH)()3). Density functional theory calculations of model alkene-substitution reactions at a diimine-palladium(0) center reveal that the palladium center reacts as a nucleophile via attack of a metal-based lone pair on the empty pi orbital of the incoming olefin. This orbital picture contrasts that of traditional ligand substitution reactions, in which the incoming ligand donates electron density into an acceptor orbital on the metal. On the basis of these results, olefin substitution at palladium(0) is classified as an "inverse-electron-demand" ligand substitution reaction. PMID- 15535710 TI - Digital photoswitching of fluorescence based on the photochromism of diarylethene derivatives at a single-molecule level. AB - Photochromic reactions of diarylethene derivatives were detected at a single molecule level by using a fluorescence technique. Fluorescent photoswitching molecules in which photochromic diarylethene and fluorescent bis(phenylethynyl)anthracene units are linked through an adamantyl spacer were synthesized, and switching of fluorescence upon irradiation with UV and visible light was followed in solution as well as on polymer films at the single-molecule level. Although in solution the fluorescence intensity gradually changed upon irradiation with UV and visible light, digital on/off switching between two discrete states was observed at the single-molecule level. The "on"- and "off" times were dependent on the power of UV and visible light. When the power of UV and visible light was increased, the average on- and off-times became short in proportion to the reciprocal power of the light. The response-times were found to show distribution. The distribution of the on- and off-times is considered to reflect the difference in the micro-environment as well as conformation of the molecules. PMID- 15535711 TI - Selective dispersion of single-walled carbon nanotubes in the presence of polymers: the role of molecular and colloidal length scales. AB - Dimensionality is known to play a key role in the solution behavior of nano- and mesoparticles. In particular, the shape and the range of the attractive van der Waals interparticle potential are determined by the number of microscopic versus mesoscopic dimensions. For single-walled nanotubes (SWNTs), where two of the dimensions are nanoscopic and one is mesoscopic, the intertube attraction is relatively short ranged, albeit very steep. The very large attraction (compared to the thermal energy, K(b)T) among long SWNTs leads to aggregation at different levels and constitutes a major barrier for manipulation and utilization of SWNTs. This study demonstrates that it is possible to shape the intertube potential by decorating SWNTs with end-tethered polymers. In good solvent conditions for the polymers, entropic repulsion among the tethered chains generates a free energy barrier that prevents SWNTs from approaching the attractive part of the intertube potential. Consequentially, stable dispersions of individual, well separated SWNTs can be prepared. Investigation of different chain lengths and tethering densities of the polymers as well as the interparticle potentials for nanometric versus mesoscopic particles suggests that polymer-induced steric stabilization provides a generic method for separation of SWNTs from mixtures of colloidal species, as demonstrated experimentally. PMID- 15535712 TI - Marked effect of aromatic solvent on unfolding rate of helical ethynylhelicene oligomer. AB - Optically active acyclic ethynylhelicene oligomers were synthesized in high yields by a two-directional method involving Sonogashira coupling and deprotection. Their CD spectra in chloroform exhibited large differences between the oligomers with less than seven helicenes and their higher homologues, which indicated the formation of helical structures for the latter and random coil structures for the former. The helical heptamer gradually unfolded to a random coil structure in chloroform at room temperature. The unfolding rate was examined by CD in several aromatic solvents as well, and the rate constant k was found to be highly dependent on the type of aromatic substituent: k differed by seven orders of magnitude between iodobenzene and trifluoromethylbenzene. Several features of the rates are notable: The reaction rates in halobenzenes were in the order of iodobenzene > bromobenzene > chlorobenzene > benzene > fluorobenzene > m difluorobenzene, those in alkylbenzenes were styrene > phenylacetylene > ethylbenzene > toluene > benzene, and those in heteroatom-substituted arenes were thioanisole > benzonitrile > anisole > ethyl benzoate > benzene > trifluoromethylbenzene. The log k values exhibited good correlation with the absolute hardness, eta, of the arenes, and higher unfolding rates were observed in the soft arenes. Vapor pressure osmometry studies indicated that the helical structure of the heptamer is dimeric in benzene, fluorobenzene, and trifluoromethylbenzene, while the random coil structure of the heptamer is monomeric in chloroform and toluene. When a chloroform solution of the random coil structure was concentrated to a small volume, the helical structure could be regenerated. PMID- 15535713 TI - Foliacenes: ab initio modeling of metallocomplexes exhibiting a unique form of 16 electron, metal-induced aromaticity. AB - Ab initio calculations predict that the cyclic trefoilenes 2 can be stabilized by formation of a complex 4 with early transition metals. The metal atom within the complex is nested within the carbon ring and is considerably closer to the ring centroid than in traditional metallocene complexes. Stabilization is explained by a unique form of 16-electron delocalization involving the metal atom, for which we suggest the name "foliate aromaticity". The aromaticity of various polyfoliate systems such as 9 suggests this 16-electron motif is more robust than Clar-like aromatic 6pi-sextets. The open hemisphere of the metal in such foliacene complexes is predicted to coordinate a variety of ligands. PMID- 15535714 TI - Properties of fullerene[50] and D5h decachlorofullerene[50]: a computational study. AB - Stimulated by the recent preparation and characterization of the first [50]fullerene derivative, decachlorofullerene[50] (Science 2004, 304, 699), we have performed a systematic density functional study on the electronic and spectroscopic properties of C(50), its anions and derivatives such as C(50)Cl(10) and C(50)Cl(12). The ground state of C(50) has D(3) symmetry with a spheroid shape, and is highly aromatic; the best D(5h)C(50) singlet is nonaromatic. Both D(3)() and D(5h)() isomers of C(50) have high electron affinities and can be reduced easily. Due to the unstable fused pentagon structural features, C(50) is chemically labile and subject to addition reactions such as chlorination, dimerization and polymerization. The equatorial pentagon-pentagon fusions of D(5h)C(50) are active sites for chemical reactions; hence, D(5h)C(50) may behave as a multivalent group. The computed IR, Raman, (13)C NMR and UV-vis spectra of the D(5h)C(50)Cl(10) molecule agree well with the experimental data. Finally, D(5h)C(50)Cl(10) is predicted to have a high electron affinity and, hence, might serve as an electron-acceptor in photonic/photovoltaic applications. The geometry and (13)C NMR chemical shifts of C(50)Cl(12) were computed to assist further isolation experiments. PMID- 15535715 TI - Catalytic mechanism of yeast cytosine deaminase: an ONIOM computational study. AB - The complete path for the deamination reaction catalyzed by yeast cytosine deaminase (yCD), a zinc metalloenzyme of significant biomedical interest, has been investigated using the ONIOM method. Cytosine deamination proceeds via a sequential mechanism involving the protonation of N(3), the nucleophilic attack of C(4) by the zinc-coordinated hydroxide, and the cleavage of the C(4)-N(4) bond. The last step is the rate determining step for the generation of the zinc bound uracil. Uracil is liberated from the Zn atom by an oxygen exchange mechanism that involves the formation of a gem-diol intermediate from the Zn bound uracil and a water molecule, the C(4)-O(Zn) cleavage, and the regeneration of the Zn-coordinated water. The rate determining step in the oxygen exchange is the formation of the gem-diol intermediate, which is also the rate determining step for the overall yCD-catalyzed deamination reaction. PMID- 15535716 TI - What makes the trifluoride anion F3 - so special? A breathing-orbital valence bond ab initio study. AB - The ground states of the F(3)(-) and H(3)(-) hypercoordinated anions are investigated and analyzed in terms of valence bond structures by means of the breathing-orbital valence bond method. While H(3)(-) is described reasonably well as the interplay of two major Lewis structures, H(2) + H(-) and its mirror image, the description of F(3)(-) requires a further structure, of the type F(*)F( )F(*), which strongly stabilizes the trimer relative to the dissociation products, and endows the F(3)(-) ground state with a predominant three-electron bond character. It follows that the simple picture that is closest to the true nature of F(3)(-) is a resonating combination of F(2)(-) + F(*) and its mirror image. This peculiarity of the F(3)(-) electronic structure is at the origin of its preferred dissociation channel leading to F(2)(-) + F(*) rather than to the most stable product F(2) + F(-), at high collision energies. The three-electron bond character of F(3)(-) is also the root cause for the failure of the Hartree Fock and density functional methods for this species, and for its strong tendency to artifactual symmetry-breaking. As an alternative to the Rundle-Pimentel model, the origins of the stability of F(3)(-), as opposed to the instability of H(3)( ), CH(5)(-), and other S(N)2 transition states, are analyzed in the framework of valence bond state correlation diagrams [Shaik, S.; Shurki, A. Angew. Chem., Int. Ed. 1999, 38, 586]. It is found that a fundamental factor of stability for X(3)( ) is the presence of lone pairs on the X fragment. The explanation carries over to other trihalide anions, and to isoelectronic 22-valence electron hypercoordinated anions. PMID- 15535717 TI - Electrochemical potential-step investigations of the aerobic interconversions of [NiFe]-hydrogenase from Allochromatium vinosum: insights into the puzzling difference between unready and ready oxidized inactive states. AB - Dynamic electrochemical studies, incorporating catalytic voltammetry and detailed potential-step manipulations, provide compelling evidence that the oxidized inactive state of [NiFe]-hydrogenases termed Unready (or Ni-A) contains a product of partial reduction of O(2) that is trapped in the active site. PMID- 15535718 TI - Redox inactivation of human 15-lipoxygenase by marine-derived meroditerpenes and synthetic chromanes: archetypes for a unique class of selective and recyclable inhibitors. AB - The selective inhibition of human 15-lipoxygenase (15-hLO) could serve as a promising therapeutic target for the prevention of atherosclerosis. A screening of marine sponges revealed that crude extracts of Psammocinia sp. exhibited potent 15-hLO inhibitory activity. Bioassay-guided fractionation led to the isolation of chromarols A-E (8-12) as potent and selective inhibitors of 15-hLO. An additional 22 structurally related compounds, including meroditerpenes from the same Psammocinia sp. (3, 4, 13-16) and our pure compound repository (17, 18), commercially available tocopherols (19-24), and synthetic chromanes (25-32), were evaluated for their ability to inhibit human lipoxygenases. The 6-hydroxychromane moiety found in chromarols A-D was identified as essential for the selective redox inhibition of 15-hLO. Furthermore, the oxidized form of the 6 hydroxychromane could be reduced by ascorbate, suggesting a potential regeneration pathway for these inhibitors in the body. This pharmacophore represents a promising paradigm for the development of a unique class of recyclable 15-hLO redox inhibitors for the treatment of atherosclerosis. PMID- 15535719 TI - FeCl3-activated oxidation of alkanes by [Os(N)O3]-. AB - Although the ion [Os(VIII)(N)(O)(3)](-) is a stable species and is not known to act as an oxidant for organic substrates, it is readily activated by FeCl(3) in CH(2)Cl(2)/CH(3)CO(2)H to oxidize alkanes efficiently at room temperature. The oxidation can be made catalytic by using 2,6-dichloropyridine N-oxide as the terminal oxidant. The active intermediates in stoichiometric and catalytic oxidation are proposed to be [(O)(3)Os(VIII)N-Fe(III)] and [Cl(4)(O)Os(VIII)N Fe(III)], respectively. PMID- 15535720 TI - Direct chemical synthesis of the beta-D-mannans: the beta-(1-->2) and beta-(1- >4) series. AB - The direct syntheses of a beta-(1-->2)-mannooctaose and of a beta-(1-->4) mannohexaose are reported by means of 4,6-O-benzylidene-protected beta-mannosyl donors. The synthesis of the (1-->2)-mannan was achieved by means of the sulfoxide coupling protocol, whereas the (1-->4)-mannan was prepared using the analogous thioglycoside/sulfinamide methodology. In the synthesis of the (1-->4) mannan, the glycosylation yields and stereoselectivities remain approximately constant with increasing chain length, whereas those for the (1-->2)-mannan consist of two groups with the formation of the tetra- and higher saccharides giving yields and selectivities consistently lower than those of the lower homologues. The decrease in yield after the trisaccharide in the (1-->2)-mannan synthesis is attributed to steric interference by the n-3 residue and is consistent with the collapsed, disordered structure predicted by early computational work. The consistently high yields and selectivities seen in the synthesis of the (1-->4)-mannan are congruent with the more open, ordered structure originally predicted for this polymer. The lack of order in the structure of the (1-->2)-mannan, as compared to the high degree of order in the (1-->4)-mannan, is also evident from a comparison of the NMR spectra of the two polymers and even from their physical nature: the (1-->2)-mannan is a gum and the (1-->4)-mannan is a high melting solid. PMID- 15535721 TI - Synthesis of Eu3+-doped core and core/shell nanoparticles and direct spectroscopic identification of dopant sites at the surface and in the interior of the particles. AB - A variety of redispersible Eu(3+)-doped LaPO(4) nanoparticles have been prepared in a high-boiling coordinating solvent mixture, and the Eu(3+) lattice sites of these materials have been investigated by luminescence line-narrowing measurements. In this spectroscopic method, Eu(3+) ions occupying different lattice sites are selectively excited with a tunable narrow-bandwidth laser system and distinguished by their luminescence spectra ("site-selective spectroscopy"). Depending on the concentration of the dopant, up to three different lattice sites could be identified in the interior of the LaPO(4) nanoparticles. These sites correspond to those known from bulk LaPO(4). In addition, a variety of surface sites is observed, which could be converted completely into bulk sites by overgrowing the nanoparticles with a shell of pure LaPO(4). The surface sites are identical to those obtained by reacting Eu(3+) with the surface of pure LaPO(4) nanoparticles. The spectroscopic properties of Eu(3+)-doped LaPO(4) nanoparticles differ from those of pure EuPO(4) nanoparticles, which were also investigated. Remarkably, the core/shell synthesis investigated in this paper allows one to prepare doped nanoparticles that contain no other dopant sites than those known from the corresponding bulk material. PMID- 15535722 TI - Highly efficient dye-sensitized solar cells with a titania thin-film electrode composed of a network structure of single-crystal-like TiO2 nanowires made by the "oriented attachment" mechanism. AB - In this study, single-crystal-like anatase TiO(2) nanowires were formed in a network structure by surfactant-assisted self-assembling processes at low temperature. The crystal lattice planes of the nanowires and networks of such wires composed of many nanoparticles were almost perfectly aligned with each other due to the "oriented attachment" mechanism, resulting in the high rate of electron transfer through the TiO(2) nanonetwork with single-crystal-like anatase nanowires. The direction of crystal growth of oriented attachment was controlled by changing the mole ratio of acetylacetone to Ti, that is, regulating both the adsorption of surfactant molecules via control of the reaction rate and the surface energy. A single-crystalline anatase exposing mainly the [101] plane has been prepared, which adsorbed ruthenium dye over 4 times higher as compared to P 25. A high light-to-electricity conversion yield of 9.3% was achieved by applying the titania nanomaterials with network structure as the titania thin film of dye sensitized solar cells. PMID- 15535723 TI - Hypervalency avoided: simple substituted BrF3 and BrF5 molecules. Structures, thermochemistry, and electron affinities of the bromine hydrogen fluorides HBrF2 and HBrF4. AB - Five different pure density functional theory (DFT) and hybrid Hartree-Fock/DFT methods have been used to search for the molecular structures, thermochemistry, and electron affinities of the bromine hydrogen fluorides HBrF(n)/HBrF(n)(-) (n = 2, 4). The basis sets used in this work are of double-zeta plus polarization quality in conjunction with s- and p-type diffuse functions, labeled as DZP++. Structures with Br-F and Br-H normal bonds, that is, HBrF(2)/HBrF(2)(-) with C(2v) or C(s) symmetry and HBrF(4)/HBrF(4)(-) with C(4v) or C(s) symmetry, are genuine minima. However, unlike the original BrF(3) and BrF(5) molecules, the global minima for HBrF(n)/HBrF(n)(-) (n = 2, 4) species are predicted to be complexes, some of which contain hydrogen bonds. The demise of the hypervalent structures is due to the availability of favorable dissociation products involving HF, which has a much larger dissociation energy than F(2). Similar reasoning suggests that PF(4)H, SF(3)H, SF(5)H, ClF(2)H, ClF(4)H, AsF(4)H, SeF(3)H, and SeF(5)H will all be hydrogen bond structures incorporating diatomic HF. The most reasonable theoretical values of the adiabatic electron affinities (EA(ad)) are 3.69 (HBrF(2)) and 4.38 eV (HBrF(4)) with the BHLYP method. These electron affinities are comparable to those of the analogous molecules: Br(2)F(n), ClBrF(n), and BrF(n)(+1) systems. The first F-atom dissociation energies for the neutral global minima are 60 (HBrF(2)) and 49 kcal/mol (HBrF(4)) with the B3LYP method. The first H-atom dissociation energies for the same systems are 109 (HBrF(2)) and 116 kcal/mol (HBrF(4)). The large Br-H bond energies are not sufficient to render the hypervalent structures energetically tenable. The dissociation energies for the complexes to their fragments are relatively small. PMID- 15535724 TI - Site-isolation effects in a dendritic nickel catalyst for the oligomerization of ethylene. AB - Dendrimers, specifically suited to construct site-isolated groups due to their well-defined hyperbranched structure, have been used as a ligand design element for the construction of nickel catalysts for ethylene oligomerization. The dendritic P,O ligand indeed suppresses the formation of inactive bis(P,O)Ni complexes in toluene, as is evident from NMR studies, and, as a consequence, outperforms the parent ligand in catalysis in this solvent. The dendritic effect observed in methanol is more subtle because both the dendritic ligand 1 and the parent 2 form bis(P,O)nickel complexes in solution according to NMR spectroscopy. Unlike the parent complex 8, the dendritic bis(P,O)Ni complex 7 derived from dendrimer ligand 1 is able to dissociate to a mono-ligated species under catalytic conditions, that is, 40 bar ethylene and 80 degrees C, which can enter the catalytic cycle. Indeed, dendritic ligand 1 gives much more active nickel catalysts for the oligomerization in methanol than does 2. PMID- 15535725 TI - Luminescence quenching of a phosphorescent conjugated polyelectrolyte. AB - The photophysical and luminescence quenching properties of a platinum(II) acetylide-based conjugated polyelectrolyte, Pt-p, which features carboxylic acid solubilizing groups are reported. The Pt-acetylide polymer is water soluble, and it exhibits phosphorescence from a triplet pi,pi exciton based on the conjugated backbone. The phosphorescence from Pt-p is quenched by viologens with different charges (MV(+), MV(2+), and MV(4+)), and in each case the quenching is dominated by a dynamic (diffusional) mechanism. Comparison of the Stern-Volmer quenching properties of Pt-p with those of a structurally analogous fluorescent organic polyelectrolyte leads to the conclusion that the amplified quenching effect, which is commonly observed for fluorescent conjugated polyelectrolytes, is not important for the platinum acetylide phosphorescent conjugated polyelectrolyte. PMID- 15535726 TI - Supramolecular polymer formation by metal-ligand complexation: Monte Carlo simulations and analytical modeling. AB - Equilibrium metal-ligand complexation leading to formation of linear or ringlike supramolecular polymers is studied by means of Monte Carlo (MC) simulations and theoretical analysis. We found that in most of the cases high-molecular-weight polymers are formed over a rather narrow composition range (near the 2:1 ligand metal ratio). Besides the imbalance in the number of metals and ligands, the molecular weight decrease in the metal-rich area is caused by an increase in 1:1 ligand-metal complex formation. The results of simulations and theoretical modeling show that the fraction of 1:1 complexes considerably decreases for metal ligand pairs with a high cooperativity of complexation. On the basis of our analytical model, we suggest a simple criterion for choosing the metal/ligand pair to achieve high molecular weight complexes in a broad range of metal-rich compositions. Dilution of a solution of metallosupramolecular polymers is found to decrease the average molecular weight and to enhance ring formation, which otherwise is very limited. PMID- 15535727 TI - Kinetics of radical heterolysis reactions forming alkene radical cations. AB - Rate constants for heterolytic fragmentation of beta-(ester)alkyl radicals were determined by a combination of direct laser flash photolysis studies and indirect kinetic studies. The 1,1-dimethyl-2-mesyloxyhexyl radical (4a) fragments in acetonitrile at ambient temperature with a rate constant of k(het) > 5 x 10(9) s( 1) to give the radical cation from 2-methyl-2-heptene (6), which reacts with acetonitrile with a pseudo-first-order rate constant of k = 1 x 10(6) s(-1) and is trapped by methanol in acetonitrile in a reversible reaction. The 1,1-dimethyl 2-(diphenylphosphatoxy)hexyl radical (4b) heterolyzes in acetonitrile to give radical cation 6 in an ion pair with a rate constant of k(het) = 4 x 10(6) s(-1), and the ion pair collapses with a rate constant of k < or = 1 x 10(9) s(-1). Rate constants for heterolysis of the 1,1-dimethyl-2-(2,2-diphenylcyclopropyl)-2 (diphenylphosphatoxy)ethyl radical (5a) and the 1,1-dimethyl-2-(2,2 diphenylcyclopropyl)-2-(trifluoroacetoxy)ethyl radical (5b) were measured in various solvents, and an Arrhenius function for reaction of 5a in THF was determined (log k = 11.16-5.39/2.3RT in kcal/mol). The cyclopropyl reporter group imparts a 35-fold acceleration in the rate of heterolysis of 5a in comparison to 4b. The combined results were used to generate a predictive scale for heterolysis reactions of alkyl radicals containing beta-mesyloxy, beta-diphenylphosphatoxy, and beta-trifluoroacetoxy groups as a function of solvent polarity as determined on the E(T)(30) solvent polarity scale. PMID- 15535728 TI - Enantioselective separation on a naturally chiral surface. AB - Kinked-stepped, high Miller index surfaces of metal crystals are chiral and, therefore, exhibit enantiospecific properties. Previous temperature-programmed desorption (TPD) spectra have shown that the desorption energies of R-3 methylcyclohexanone (R-3-MCHO) on the chiral Cu(643)(R) and Cu(643)(S) surfaces are enantiospecific (J. Am. Chem. Soc. 2002, 124, 2384). Here, a comparison of the TPD spectra from Cu(111), Cu(221), Cu(533), Cu(653)(R&S), and Cu(643)(R&S) surfaces reveals that the enantiospecific desorption occurs from the chiral kink sites on the Cu(643) surfaces. Titration of the chiral kink sites with I atoms confirms this assignment of desorption features in the TPD spectra. Finally, the enantiospecific difference in the desorption energies of R- and S-3-MCHO has been used as the basis for demonstration of an enantioselective, kinetic separation of racemic 3-MCHO into its purified components during adsorption and desorption on the Cu(643)(R&S) surfaces. PMID- 15535733 TI - Kinetics of protoporphyrin IX formation in rat oral mucosa and skin after application of 5-aminolevulinic acid and its methylester. AB - The kinetics of accumulation of protoporphyrin IX (PpIX) after topical application of 5-aminolevulinic acid (ALA) and its methylester (5-aminolevulinic acid methylester [ALA-Me]) was studied on rat oral mucosa. The accumulation of PpIX in mucosa and skin after intravenous injection of ALA and ALA-Me was also studied. The elimination rate of PpIX was dependent on drug and dose as well as on administration route. Application of ALA on rat oral mucosa and skin caused a systemic effect with PpIX building up in remote skin sites not exposed to the drugs. No such systemic effect was seen after application of ALA-Me either in mucosa or on skin. Intravenous injection of the drugs (0.2 g/kg) leads to more fluorescence in the skin than topical application of the drug (20%). For mucosa, the opposite is true. Maximal PpIX fluorescence appeared later after application of high concentrations of the drugs (around 8 h for 5% and 20% wt/wt) than after application of low concentrations (around 3-5 h for 1% and 2% wt/wt). PMID- 15535734 TI - UV exposure, genetic targets in melanocytic tumors and transgenic mouse models. AB - The genetic changes and corruption of kinase activity in melanomas appear to revolve around a central axis: mitogenic signaling along the RAS pathway down to transcription regulation by pRB. Epidemiological studies point to the importance of ultraviolet (UV) radiation in the etiology of melanoma, but where and how UV radiation is targeted to contribute to the oncogenic signaling remains obscure. Animal models of melanoma genesis could serve to clarify this issue, but many of these models are not responsive to UV exposure. Most interesting advances have been made by using transgenic mice that carry genetic defects that are known to be relevant to human melanoma: specifically, dysfunction in the tumor suppressive action of p16INK4a or a receptor tyrosine kinase/RAS pathway, that is constitutively activated in melanocytes. The latter types of mice appear to be most responsive to (neonatal) UV exposure. Whether this is due to a general increase in target cells by melanocytosis and a paucity or complete lack of pigment, or a possible UV-induced response of the promoter-enhancer of the transgene or a genuinely independent and additional genetic alteration caused by UV exposure needs to be established. Importantly, the full effect of UV radiation needs to be ascertained in mice with different pigmentation by varying the wavelengths, UV-B versus UV-A1, and the exposure schedules, i.e. neonatal versus adult and chronic versus intermittent overexposure. Intermittent UV-B overexposure deserves special attention because it most strongly evokes proliferative responses in melanocytes. PMID- 15535735 TI - Photochemistry and phototoxicity of fluocinolone 16,17-acetonide. AB - Fluocinolone 16,17-acetonide is a corticosteroid used topically to treat various inflammatory skin diseases. Its photoreactivity was studied under UV-A and UV-B light in aqueous buffer in the presence of oxygen. This drug is photolabile under UV-B light and, to a lesser extent, under UV-A light, which is absorbed far less. In phosphate buffer, approximately 80% of fluocinolone acetonide decomposes after 5 J/cm2 of UV-B irradiation, whereas under 30 J/cm2 of UV-A light approximately only 20% decomposes. Both the drug and its photoproducts have been evaluated through a battery of in vitro studies and found to cause photohemolysis and induce photodamage to proteins (erythrocyte ghosts, bovine serum albumin) and linoleic acid. In addition, one of the photoproducts (the 17-hydroperoxy derivative) is highly toxic in the dark. Therefore, both loss of therapeutic activity and light-induced adverse effects may be expected when patients expose themselves to sunlight after drug administration. A major mechanism for phototoxicity involves radicals forming from drug breakdown, at least under UV-B, although reactive oxygen species may play a role, particularly under UV-A. PMID- 15535737 TI - Enhancement of laser cancer treatment by a chitosan-derived immunoadjuvant. AB - A chitosan derivative, glycated chitosan (GC), has been used as an immunostimulant for cancer treatment in laser immunotherapy. The function of GC is to enhance the host immune response after direct cancer cell destruction by a selective laser photothermal interaction. To further test its effects, laser immunotherapy was extended to include several different adjuvants for immunological stimulation and to include photodynamic therapy (PDT) as a different tumor-destruction mechanism. Complete Freund (CF) adjuvant, incomplete Freund (IF) adjuvant and Corynebacterium parvum (CP) were selected for treatment of metastatic mammary tumors in rats, in combination with a selective photothermal interaction. The solution of the immunoadjuvants admixed with indocyanine green (ICG), a light-absorbing dye, was injected directly into the tumors, followed by noninvasive irradiation of an 805 nm laser. Combined with PDT, in the treatment of tumors in mice, GC was administered peritumorally immediately after laser irradiation. The survivals of treated animals were compared with untreated control animals. In the treatment of rat tumors, CF, IF and CP raised the cure rates from 0% to 18%, 7% and 9%, respectively. In comparison, GC resulted in a 29% long-term survival. In the treatment of EMT6 mammary sarcoma in mice, GC of 0.5% and 1.5% concentrations increased the cure rates of Photofrin-based PDT treatment from 38% to 63% and 75%, respectively. In the treatment of Line 1 lung adenocarcinoma in mice, a 1.67% GC solution enabled a noncurative meso-substituted tetra(meta-hydroxy-phenyl)chlorin-based PDT to cure 37% of the tumor-bearing mice. The experimental results of this study confirmed our previous studies, showing that immunoadjuvants played an active role in laser-related cancer treatment and that GC significantly enhanced the efficacy of laser cancer treatment. PMID- 15535736 TI - Optical properties of human prostate at 732 nm measured in mediated photodynamic therapy. AB - Characterization of the tissue light penetration in prostate photodynamic therapy (PDT) is important to plan the arrangement and weighting of light sources so that sufficient light fluence is delivered to the treatment volume. The optical properties (absorption [mu(a)], transport scattering [mu(s)'] and effective attenuation [mu(eff)] coefficients) of 13 patients with locally recurrent prostate cancer were measured in situ using interstitial isotropic detectors. Measurements were made at 732 nm before and after motexafin lutetium (MLu) mediated PDT in four quadrants. Optical properties were derived by applying the diffusion theory to the fluence rates measured at several distances (0.5-5 cm) from a point source. mu(a) and mu(s)' varied between 0.07 and 1.62 cm(-1) (mean 0.37 +/- 0.24 cm(-1)) and 1.1 and 44 cm(-1) (mean 14 +/- 11 cm(-1)), respectively. mu(a) was proportional to the concentration of MLu measured by an ex vivo fluorescence assay. We have observed, on average, a reduction of the MLu concentration after PDT, presumably due to the PDT consumption of MLu. mu(eff) varied between 0.91 and 6.7 cm(-1) (mean 2.9 +/- 0.7 cm(-1)), corresponding to an optical penetration depth (delta = 1/micro(eff)) of 0.1-1.1 cm (mean 0.4 +/- 0.1 cm). The mean penetration depth at 732 nm in human prostate is at least two times smaller than that found in normal canine prostates, which can be explained by a four times increase of the mean value of mu(s)' in human prostates. The mean light fluence rate per unit source strength at 0.5 cm from a point source was 1.5 +/- 1.1 cm(-2), excluding situations when bleeding occurs. The total number of measurements was N = 121 for all mean quantities listed above. This study showed significant inter- and intraprostatic differences in the optical properties, suggesting that a real-time dosimetry measurement and feedback system for monitoring light fluences during treatment should be considered for future PDT studies. PMID- 15535738 TI - Endogenous fluorescence spectroscopy of cell suspensions for chemopreventive drug monitoring. AB - Cancer chemopreventive agents such as N-4-(hydroxyphenyl)retinamide (4HPR) are thought to prevent cancers by suppressing growth or inducing apoptosis in precancerous cells. Mechanisms by which these drugs affect cells are often not known, and the means to monitor their effects is not available. In this study endogenous fluorescence spectroscopy was used to measure metabolic changes in response to treatment with 4HPR in ovarian and bladder cancer cell lines. Fluorescence signals consistent with nicotinamide adenine dinucleotide (NADH), flavin adenine dinucleotide (FAD) and tryptophan were measured to monitor cellular activity through redox status and protein content. Cells were treated with varying concentrations of 4HPR and measured in a stable environment with a sensitive fluorescence spectrometer. Results suggest that redox signal of all cells changed in a similar dose-dependant manner but started at different baseline levels. Redox signal changes depended primarily on changes consistent with NADH fluorescence, whereas the FAD fluorescence remained relatively constant. Similarly, tryptophan fluorescence decreased with increased drug treatment, suggesting a decrease in protein production. Given that each cell line has been shown to have a different apoptotic response to 4HPR, fluorescence redox values along with changes in tryptophan fluorescence may be a response as well as an endpoint marker for chemopreventive drugs. PMID- 15535740 TI - Consolidation of long-term memory: evidence and alternatives. AB - Memory loss in retrograde amnesia has long been held to be larger for recent periods than for remote periods, a pattern usually referred to as the Ribot gradient. One explanation for this gradient is consolidation of long-term memories. Several computational models of such a process have shown how consolidation can explain characteristics of amnesia, but they have not elucidated how consolidation must be envisaged. Here findings are reviewed that shed light on how consolidation may be implemented in the brain. Moreover, consolidation is contrasted with alternative theories of the Ribot gradient. Consolidation theory, multiple trace theory, and semantization can all handle some findings well but not others. Conclusive evidence for or against consolidation thus remains to be found. PMID- 15535741 TI - Developmental dyslexia and specific language impairment: same or different? AB - Developmental dyslexia and specific language impairment (SLI) were for many years treated as distinct disorders but are now often regarded as different manifestations of the same underlying problem, differing only in severity or developmental stage. The merging of these categories has been motivated by the reconceptualization of dyslexia as a language disorder in which phonological processing is deficient. The authors argue that this focus underestimates the independent influence of semantic and syntactic deficits, which are widespread in SLI and which affect reading comprehension and impair attainment of fluent reading in adolescence. The authors suggest that 2 dimensions of impairment are needed to conceptualize the relationship between these disorders and to capture phenotypic features that are important for identifying neurobiologically and etiologically coherent subgroups. PMID- 15535742 TI - Conscientiousness and health-related behaviors: a meta-analysis of the leading behavioral contributors to mortality. AB - Previous research has established conscientiousness as a predictor of longevity (H. S. Friedman et al., 1993; L. R. Martin & H. S. Friedman, 2000). To better understand this relationship, the authors conducted a meta-analysis of conscientiousness-related traits and the leading behavioral contributors to mortality in the United States (tobacco use, diet and activity patterns, excessive alcohol use, violence, risky sexual behavior, risky driving, suicide, and drug use). Data sources were located by combining conscientiousness-related terms and relevant health-related behavior terms in database searches as well as by retrieving dissertations and requesting unpublished data from electronic mailing lists. The resulting database contained 194 studies that were quantitatively synthesized. Results showed that conscientiousness-related traits were negatively related to all risky health-related behaviors and positively related to all beneficial health-related behaviors. This study demonstrates the importance of conscientiousness' contribution to the health process through its relationship to health-related behaviors. PMID- 15535743 TI - Timing of pubertal maturation in girls: an integrated life history approach. AB - Life history theory provides a metatheoretical framework for the study of pubertal timing from an evolutionary-developmental perspective. The current article reviews 5 middle-level theories--energetics theory, stress-suppression theory, psychosocial acceleration theory, paternal investment theory, and child development theory--each of which applies the basic assumptions of life history theory to the question of environmental influences on timing of puberty in girls. These theories converge in their conceptualization of pubertal timing as responsive to ecological conditions but diverge in their conceptualization of (a) the nature, extent, and direction of environmental influences and (b) the effects of pubertal timing on other reproductive variables. Competing hypotheses derived from the 5 perspectives are evaluated. An extension of W. T. Boyce and B. J. Ellis's (in press) theory of stress reactivity is proposed to account for both inhibiting and accelerating effects of psychosocial stress on timing of pubertal development. This review highlights the multiplicity of (often unrecognized) perspectives guiding research, raises challenges to virtually all of these, and presents an alternative framework in an effort to move research forward in this arena of multidisciplinary inquiry. PMID- 15535744 TI - The role of representative design in an ecological approach to cognition. AB - Egon Brunswik argued that psychological processes are adapted to environmental properties. He proposed the method of representative design to capture these processes and advocated that psychology be a science of organism-environment relations. Representative design involves randomly sampling stimuli from the environment or creating stimuli in which environmental properties are preserved. This departs from systematic design. The authors review the development of representative design, examine its use in judgment and decision-making research, and demonstrate the effect of design on research findings. They suggest that some of the practical difficulties associated with representative design may be overcome with modern technologies. The importance of representative design in psychology and the implications of this method for ecological approaches to cognition are discussed. PMID- 15535745 TI - Birth weight and cognitive ability in childhood: a systematic review. AB - Individual differences in cognitive ability may in part have prenatal origins. In high-risk (low birth weight/premature) babies, birth weight correlates positively with cognitive test scores in childhood, but it is unclear whether this holds for those with birth weights in the normal range. The authors systematically reviewed literature on the relationship between normal birth weight (more than 2,500 g) and childhood intelligence in term (37-42-week gestation) deliveries. Six studies met the inclusion criteria, and the authors present a comprehensive narrative review of these studies. There was a small, consistent, positive association between birth weight and childhood cognitive ability, even when corrected for confounders. Parental social class accounted for a larger proportion of the variance than birth weight, and these 2 variables were largely independent. PMID- 15535746 TI - Caregiving representations of mothers of behaviorally inhibited and uninhibited preschool children. AB - This study examined differences in the caregiving representations of mothers of 3 to 4-year-old behaviorally inhibited and uninhibited children with secure or insecure attachments. Mothers of inhibited children perceived their children as more vulnerable than did mothers of uninhibited children, and they acknowledged difficulties associated with their children's inhibited temperament. However, mothers of insecure inhibited children were less likely than mothers of secure inhibited children to validate their children's emotional experiences and to be aware of their children's internal states and perspectives, and they showed higher levels of boundary violation and defense against negative affect. Implications of the more problematic caregiving representations of mothers of insecure inhibited children for parent-child relationships are discussed. PMID- 15535748 TI - In the eye of the beholder: subjective and observer ratings of middle-class African American mother-adolescent interactions. AB - Middle-class African American mothers and adolescents (n=81) participated in a dyadic interaction task in early adolescence (M=13.06 years, SD=1.27) and then again 2 years later (M=15.01 years, SD=1.27). Following the task, mothers and adolescents rated their own and their partner's support and involvement in the task; observers rated videotaped interactions for the same dimensions. African American mothers perceived adolescents' behavior in the interactions more positively than did observers, and both mothers and observers viewed mothers' behavior as more positive than adolescents' behavior. With observers' ratings controlled, early adolescents' more negative perceptions of their mothers' interactions led to less trust and communication with mothers 2 years later, but subjective understanding did not influence later ratings of conflict intensity. PMID- 15535747 TI - Prediction of children's empathy-related responding from their effortful control and parents' expressivity. AB - In this study, the linear and interactive relations of children's effortful control and parents' emotional expressivity to children's empathy-related responses were examined. Participants were 214 children, 4.5 to 8 years old. Children's effortful control was negatively related to their personal distress and was positively related to their sympathy. Parents' positive expressivity was marginally negatively related to children's personal distress and was marginally positively related to children's dispositional sympathy. Parents' negative expressivity was positively related to children's personal distress, but primarily at high levels of children's effortful control. Moreover, parents' negative expressivity was negatively related to children's situational sympathy at low levels of effortful control but was positively related to children's dispositional sympathy at high levels of effortful control. There were also quadratic relations between the measures of parents' expressivity and children's empathy-related responses. PMID- 15535749 TI - An experimental analysis of dynamic hypotheses about cognitive abilities and achievement from childhood to early adulthood. AB - This study examined the dynamics of cognitive abilities and academic achievement from childhood to early adulthood. Predictions about time-dependent "coupling" relations between cognition and achievement based on R. B. Cattell's (1971, 1987) investment hypothesis were evaluated using linear dynamic models applied to longitudinal data (N=672). Contrary to Cattell's hypothesis, a first set of findings indicated that fluid and crystallized abilities, as defined by the Woodcock-Johnson Psycho-Educational Battery-Revised (WJ-R; R. W. Woodcock & M. B. Johnson, 1989-1990), were not dynamically coupled with each other over time. A second set of findings provided support for the original predictions and indicated that fluid ability was a leading indicator of changes in achievement measures (i.e., quantitative ability and general academic knowledge). The findings of this study suggest that the dynamics of cognitive abilities and academic achievement follow a more complex pattern than that specified by Cattell's investment hypothesis. PMID- 15535750 TI - Acting and planning on the basis of a false belief: its effects on 3-year-old children's reasoning about their own false beliefs. AB - This study examined 3-year-olds' explanations for actions of theirs that were premised on a false belief. In Experiment 1, children stated what they thought was inside a crayon box. After stating "crayons," they went to retrieve some paper to draw on. Children were then shown that the box contained candles and were asked to (a) state their initial belief and (b) explain their action of getting paper. Children who were unable to retrieve their false belief were unable to correctly explain their action. Experiments 2 and 3 ruled out several alternative interpretations for these findings. In Experiment 4, children planned and acted on their false belief. Again, children who were unable to retrieve their false belief were unable to correctly explain their action. PMID- 15535751 TI - Maternal affection moderates the impact of psychological control on a child's mathematical performance. AB - This study investigated the extent to which mothers' psychological control predicts their children's mathematical performance during the children's transition from preschool to primary school over and above the impact of maternal affection and behavioral control. Also investigated was the extent to which maternal affection and behavioral control moderate the impact of mothers' psychological control. Children 5-6 years old at baseline (N=196) were followed up 6 times to measure their performance in mathematics over a 3-year period from preschool to 2nd grade. Mothers were asked to fill in a questionnaire measuring their parenting styles once every year over the 3-year period. A high level of psychological control exercised by mothers predicted their children's slow progress in mathematics. However, this impact was particularly evident among those children whose mothers reported a high level of affection. No evidence was found that children's mathematical performance had any effect on their mothers' parenting styles. PMID- 15535752 TI - Mother-child discourse in two contexts: links with child temperament, attachment security, and socioemotional competence. AB - The goal of this study was to examine whether attachment security and child temperament predicted differences in the elaboration and emotional content of mother-child discourse in 2 contexts and whether those differences were related to a child's socioemotional development. Fifty-one preschool children and their mothers were videotaped reading a storybook and discussing the child's previous behavior. These conversations were coded for maternal elaboration and emotional content. Mothers also completed reports of child temperament, attachment security, and social behavior. Children completed measures of emotional understanding, behavioral internalization, and representations of relationships. The findings suggested that both aspects of mother-child discourse were related to attachment, temperament, and socioemotional competence, although the findings varied depending on the context of the discourse. PMID- 15535753 TI - A meta-analytic review of gender variations in children's language use: talkativeness, affiliative speech, and assertive speech. AB - Three sets of meta-analyses examined gender effects on children's language use. Each set of analyses considered an aspect of speech that is considered to be gender typed: talkativeness, affiliative speech, and assertive speech. Statistically significant average effect sizes were obtained with all three language constructs. On average, girls were slightly more talkative and used more affiliative speech than did boys, whereas boys used more assertive speech than did girls. However, the average effect sizes were either negligible (talkativeness, d=0.11; assertive speech, d=0.11) or small (affiliative speech, d=0.26). Larger effect sizes were indicated for some language constructs depending on either the operational definition of the language measure, the method of recording, the child's age level, the interaction partner (adult or peer), group size, gender composition, observational setting, or type of activity. The results are interpreted in relation to social-developmental and social-constructionist approaches to gender; these views are presented as complementary--rather than competing--meta-theoretical viewpoints. PMID- 15535754 TI - Child personality and parental behavior as moderators of problem behavior: variable- and person-centered approaches. AB - Parenting x Child Personality interactions in predicting child externalizing and internalizing behavior were investigated in a variable-centered study and a person-centered study. The variable-centered study used data from a 3-year longitudinal study of 600 children 7 to 15 years old at Time 1 and 512 children 10 to 18 years old at Time 2. Parents rated child personality (five factor model), negative control, positive parenting, and child problem behavior, whereas children rated parental behavior. Hierarchical moderated regression analyses showed significant Parenting x Child Personality (benevolence and conscientiousness) interactions, principally for externalizing behavior. The interactions were largely replicable across informants and across time. The person-centered study, which classified participants into 3 types, showed that negative parental control was more related to externalizing behavior for undercontrollers than for resilients. Negative parental control enhanced internalizing behavior for overcontrollers. PMID- 15535755 TI - The limits of child effects: evidence for genetically mediated child effects on corporal punishment but not on physical maltreatment. AB - Research on child effects has demonstrated that children's difficult and coercive behavior provokes harsh discipline from adults. Using a genetically sensitive design, the authors tested the limits of child effects on adult behavior that ranged from the normative (corporal punishment) to the nonnormative (physical maltreatment). The sample was a 1994-1995 nationally representative birth cohort of 1,116 twins and their families who participated in the Environmental Risk Longitudinal Study. Results showed that environmental factors accounted for most of the variation in corporal punishment and physical maltreatment. However, corporal punishment was genetically mediated in part, and the genetic factors that influenced corporal punishment were largely the same as those that influenced children's antisocial behavior, suggesting a child effect. The authors conclude that risk factors for maltreatment are less likely to reside within the child and more likely to reside in characteristics that differ between families. PMID- 15535756 TI - More gestures than answers: children learning about balance. AB - This research extends the range of domains within which children's gestures are found to play an important role in learning. The study involves children learning about balance, and the authors locate children's gestures within a relevant model of cognitive development--the representational redescription model (A. Karmiloff Smith, 1992). The speech and gestures of children explaining a balance task were examined. Approximately one third of the children expressed one idea in speech and another in gesture. These children made significantly more learning gains than children whose gestures and speech matched. Children's gestures were an indicator, at pretest, of readiness to learn and of cognitive gains. The authors conclude that children's gestures provide crucial insight into their cognitive state and illuminate the process of learning and representational change. PMID- 15535757 TI - Infants' vagal regulation in the still-face paradigm is related to dyadic coordination of mother-infant interaction. AB - The authors investigated relations between mother-infant dyadic coordination and infants' physiological responses. Mothers (N=73) and 3-month-old male and female infants were observed in the still-face paradigm, and mothers' and infants' affective states were coded at 1-s intervals. Synchrony and levels of matching between mother-infant affective states were computed, and infants' heart rate and vagal tone were measured. Infants showed increased negative affect and heart rate and decreased vagal tone during mothers' still-face, indicating physiological regulation of distress. Infants who did not suppress vagal tone during the still face (nonsuppressors) showed less positive affect, higher reactivity and vagal suppression in normal play and reunion episodes, and lower synchrony in normal play with mothers. The results indicate that infants' physiological regulation in social interaction differs in relation to dyadic coordination of affective behaviors. PMID- 15535758 TI - Maternal preconceptions about parenting predict child temperament, maternal sensitivity, and children's empathy. AB - This study examined the influence of maternal preconceptions on child difficult temperament at 6 months and maternal sensitivity at 12-15 months and whether all 3 variables predicted children's empathy at 21-24 months. Within a low-income, ethnically diverse sample of 175 mother-child dyads, path models were tested with 3 empathy indices (prosocial, indifference, inquisitive) as outcomes. Results indicated that maternal preconceptions significantly predicted child difficult temperament, maternal sensitivity, and children's empathy. Temperament mediated the link between maternal preconceptions and inquisitiveness, and maternal sensitivity mediated the link between preconceptions and prosocial responses. Group modeling techniques revealed no significant differences across gender or ethnicity. Correlations suggested contextual effects based on the familiarity of the person in distress. The implications and utility of developing parenting interventions are discussed. PMID- 15535759 TI - Are eyes windows to a deceiver's soul? Children's use of another's eye gaze cues in a deceptive situation. AB - Three experiments examined 3- to 5-year-olds' use of eye gaze cues to infer truth in a deceptive situation. Children watched a video of an actor who hid a toy in 1 of 3 cups. In Experiments 1 and 2, the actor claimed ignorance about the toy's location but looked toward 1 of the cups, without (Experiment 1) and with (Experiment 2) head movement. In Experiment 3, the actor provided contradictory verbal and eye gaze clues about the location of the toy. Four- and 5-year-olds correctly used the actor's gaze cues to locate the toy, whereas 3-year-olds failed to do so. Results suggest that by 4 years of age, children begin to understand that eye gaze cues displayed by a deceiver can be informative about the true state of affairs. PMID- 15535760 TI - Executive function and theory of mind: stability and prediction from ages 2 to 3. AB - Several studies have demonstrated a relation between executive functioning (EF) and theory of mind (ToM) in preschoolers, yet the developmental course of this relation remains unknown. Longitudinal stability and EF-ToM relations were examined in 81 children at 24 and 39 months. At Time 1, EF was unrelated to behavioral measures of ToM but was significantly related to parent report of children's internal-state language, independent of vocabulary size. At Time 2, behavioral batteries of EF and ToM were significantly related (r=.50, p<.01). Furthermore, EF (Time 1) significantly predicted ToM (Time 2), independent of several controls. A reciprocal relation (internal-state language --> EF at Time 2) was nonsignificant with the controls included. Individual differences in EF were relatively stable. PMID- 15535761 TI - Infant and maternal behaviors regulate infant reactivity to novelty at 6 months. AB - Three issues were investigated: (a) the regulatory effects of presumed infant and maternal regulation behaviors on infant distress to novelty at 6 months, (b) stability of infant regulatory effects across contexts that vary in maternal involvement, and (c) associations and temporal dynamics between infant and maternal regulation behaviors. Participants were 87 low-risk infants and their mothers, observed at 6 months postpartum during infant exposure to novel toys. Contingencies derived from sequential analyses demonstrate that, by 6 months, some infants reduce their own distress to novelty by looking away from the novel toy or self-soothing, maternal engagement and support have comparable effects, and certain infant and maternal behaviors co-occur. Moreover, infants whose mothers engaged contingently when they looked away from the novel toy expressed less distress than comparable infants whose mothers did not. These findings implicate both infants and mothers in the development of emotion regulation during the infant's first year. PMID- 15535762 TI - Parent-infant synchrony and the social-emotional development of triplets. AB - To study the social-emotional development of triplets, 23 sets of triplets, 23 sets of twins, and 23 singleton infants (N=138) were followed from birth to 2 years. Maternal depression and social support were assessed in the postpartum period, mother-infant and father-infant interaction and the home environment were observed at 3 months, a separation-reunion episode and a maternal interview were conducted at 12 months, and infant behavior problems were evaluated at 24 months. Lower parent-infant synchrony was observed for triplets. Triplets showed less distress during maternal separation and less approach at reunion. Mothers reported lower adjustment and differentiation among siblings for triplets than for twins. Higher internalizing problems were reported for triplets, and the triplet with intrauterine growth retardation showed the poorest outcomes. Behavior problems were predicted by medical risk, maternal depression, parent infant synchrony, infant approach, and mother adjustment. Discussion focuses on developmental risk when the exclusivity of the parent-infant relationship is compromised. PMID- 15535763 TI - Adolescents' performance on the Iowa Gambling Task: implications for the development of decision making and ventromedial prefrontal cortex. AB - Healthy adolescents (79 girls, 66 boys), ages 9-17, completed the Iowa Gambling Task (IGT; A. Bechara, A. R. Damasio, H. Damasio, & S. W. Anderson, 1994) as well as working memory (digit span) and behavioral inhibition (go/no-go) tasks. Cross sectional age-related changes were seen on all 3 tasks. Gender differences were seen in IGT deck preference and attentional variables (i.e., go/no-go hit rate and forward digit span). After age, gender, and general intellectual abilities were controlled for, IGT performance was not predicted by working memory or behavioral inhibition scores. Findings suggest that the ventromedial prefrontal cortex or its connections are functionally maturing during adolescence in a manner that can be distinguished from maturation of other prefrontal regions. Development of these functions may continue into young adulthood. PMID- 15535764 TI - Beyond the individual: the impact of ethnic context and classroom behavioral norms on victims' adjustment. AB - With a sample of 1,630 sixth-grade students from 77 classrooms, the authors used hierarchical linear modeling to examine how ethnicity within context and classroom social disorder influenced the association between peer victimization and social-psychological adjustment (loneliness and social anxiety). Victimized students in classrooms where many classmates shared their ethnicity reported feeling the most loneliness and social anxiety. Additionally, classroom-level social disorder served as a moderator such that the association between victimization and anxiety was stronger in classrooms with low social disorder. Both findings were interpreted as evidence that problem behavior deviating from what is perceived as normative in a particular context heightens maladjustment. The authors discuss implications for studying ethnicity and classroom behavioral norms as context variables in peer relations. PMID- 15535765 TI - The characteristics and correlates of fantasy in school-age children: imaginary companions, impersonation, and social understanding. AB - Past research with 152 preschoolers found that having an imaginary companion or impersonating an imaginary character was positively correlated with theory of mind performance. Three years later, 100 children from this study were retested to assess the developmental course of play with imaginary companions and impersonation of imaginary characters and how these types of role play were related to emotion understanding, self-perception, and personality. The results showed that school-age children interact with imaginary companions and impersonate imaginary characters as much as preschoolers. Overall, 65% of children up to the age of 7 had imaginary companions at some point during their lives. School-age children who did not impersonate scored lower on emotion understanding. Theory of mind at age 4 predicted emotion understanding 3 years later. PMID- 15535766 TI - Genetic and environmental influences on pubertal development: longitudinal data from Finnish twins at ages 11 and 14. AB - To study sources of individual differences in pubertal development, the authors fit a sex-limitation common factor model to data reported, at ages 11 and 14 years, by 1,891 twin pairs on items that comprise the Pubertal Development Scale (PDS; A. C. Petersen, L. Crockett, M. Richards, & A. Boxer, 1988). The model divides variation into a general pubertal factor and item-specific variation and, in addition, decomposes it into constituent sources. In both boys and girls, genetic influences made the largest contribution to variance common to PDS items. Genetic and nonshared environmental factors accounted for variation specific to PDS items in boys, whereas for girls, common environmental influences were added for growth spurt and menarcheal status. For both common and item-specific variation, genetic effects were partially sex specific. Subsidiary analyses found accelerated maturation in both boys and girls who at age 14 were reared in father absent homes. PMID- 15535767 TI - On the development of the mental number line: more, less, or never holistic with increasing age? AB - Magnitude is assumed to be represented along a holistic mental number line in adults. However, the authors recently observed a unit-decade compatibility effect for 2-digit numbers that is inconsistent with this "holisticness" assumption (H. C. Nuerk, U. Weger, & K. Willmes, 2001). This study used the compatibility effect to examine whether the mental number line representation of magnitude changes toward greater or less holisticness in children from Grades 2-5. The results indicate that decades and units of 2-digit numbers are processed separately rather than holistically from Grade 2 on. However, this separate processing seems to develop from a more sequential (left-to-right) to a more parallel processing mode. Moreover, children may use different strategies depending on task demands. The results are interpreted in the framework of Siegler's overlapping waves model. PMID- 15535768 TI - The affective structure of supportive parenting: depressive symptoms, immediate emotions, and child-oriented motivation. AB - This study investigated the maternal concerns and emotions that may regulate one form of sensitive parenting, support for children's immediate desires or intentions. While reviewing a videotape of interactions with their 1-year-olds, mothers who varied on depressive symptoms reported concerns and emotions they had during the interaction. Emotions reflected outcomes either to children (child oriented concerns) or to mothers themselves (parent-oriented concerns). Child oriented concerns were associated with fewer negative emotions and more supportive behavior. Supportive parenting was high among mothers who experienced high joy and worry and low anger, sadness, and guilt. However, relations depended on whether emotions were child or parent oriented: Supportive behavior occurred more when emotions were child oriented. In addition, as depressive symptoms increased, mothers reported fewer child-oriented concerns, fewer child-oriented positive emotions, and more parent-oriented negative emotions. They also displayed less supportive behavior. Findings suggest that support for children's immediate intentions may be regulated by parents' concerns, immediate emotions, and depressive symptoms. PMID- 15535769 TI - Developmental change in feedback processing as reflected by phasic heart rate changes. AB - Heart rate was recorded from 3 age groups (8-10, 12, and 20-26 years) while they performed a probabilistic learning task. Stimuli had to be sorted by pressing a left versus right key, followed by positive or negative feedback. Adult heart rate slowed following negative feedback when stimuli were consistently mapped onto the left or right key (response-dependent condition) but not when responses to stimuli were always followed by negative feedback (uninformative condition). Young children's heart rate showed slowing following both response-dependent and uninformative negative feedback. These findings suggest that the ability to assess the relevance of performance feedback improves with age, resulting in improved adjustment to dynamical changes in the task environment. PMID- 15535770 TI - Concordance in attachment states of mind and styles with respect to fathers and mothers. AB - It is believed that by adulthood, independent attachments to the mother and the father coalesce into a single state of mind with respect to attachment. If true, states of mind with respect to mothers and fathers should be concordant. Fifty six young adults were administered two versions of the Adult Attachment Interview, each of which asked about their relationship with one parent. State of mind with respect to the father was significantly related to state of mind with respect to the mother, as were attachment styles regarding the two parents. Perceptions of attachment styles were not very related to corresponding states of mind but were related to inferred loving from a parent. PMID- 15535771 TI - Liking is for doing: the effects of goal pursuit on automatic evaluation. AB - Findings from 3 experiments suggest that participants who were actively engaged in goal pursuit, compared with those who were not pursuing the goal, automatically evaluated goal-relevant objects as relatively more positive than goal-irrelevant objects. In Experiment 3, participants' automatic evaluations also predicted their behavioral intentions toward goal-relevant objects. These results suggest the functional nature of automatic evaluation and are in harmony with the classic conceptualization of thinking and feeling as being in the service of "doing" (e.g., S. T. Fiske, 1992; W. James, 1890; K. Lewin, 1926) as well as with more recent work on the cognitive mechanics of goal pursuit (e.g., G. B. Moskowitz, 2002; J. Y. Shah & A.W. Kruglanski, 2002). PMID- 15535772 TI - Strategies for social inference: a similarity contingency model of projection and stereotyping in attribute prevalence estimates. AB - Most models of how perceivers infer the widespread attitudes and qualities of social groups revolve around either the self (social projection, false consensus) or stereotypes (stereotyping). The author suggests people rely on both of these inferential strategies, with perceived general similarity moderating their use, leading to increased levels of projection and decreased levels of stereotyping. Three studies featuring existing individual differences in perceived similarity as well as manipulated perceptions supported the predictions, with similarity yielding increased projection to, and decreased stereotyping of, various in groups and out-groups. Evidence that projection and stereotyping may serve as inferential alternatives also emerged. The model and accompanying results have implications for research on social comparison and projection, stereotyping and prejudice, and social inference. PMID- 15535773 TI - Think different: the merits of unconscious thought in preference development and decision making. AB - The role of unconscious and conscious thought in decision making was investigated in 5 experiments. Because of the low processing capacity of consciousness, conscious thought was hypothesized to be maladaptive when making complex decisions. Conversely, unconscious thought was expected to be highly effective. In Experiments 1-3, participants were presented with a complex decision problem in which they had to choose between various alternatives, each with multiple attributes. Some participants had to make a decision immediately after being presented with the options. In the conscious thought condition, participants could think about the decision for a few minutes. In the unconscious thought condition, participants were distracted for a few minutes and then indicated their decision. Throughout the experiments, unconscious thinkers made the best decisions. Additional evidence obtained in Experiments 4 and 5 suggests that unconscious thought leads to clearer, more polarized, and more integrated representations in memory. PMID- 15535774 TI - Cognitive processes in stereotype formation: the role of correct contingency learning for biased group judgments. AB - Three studies investigated contingency learning and stereotype formation in a scenario about group membership and behavior with a confounding context factor. The studies tested predictions from theoretical accounts of biased group judgments in terms of simplistic reasoning, parallel distributed memory, and pseudocontingencies. Study 1 revealed a positive correlation between erroneous stereotype formation and learning of the true contingencies with the confounding factor. Study 2 showed that a focus manipulation during encoding moderated the correlation between stereotype formation and contingency learning but not the strength of the erroneous stereotype. Study 3 used a quasiexperimental comparison between participants with biased versus unbiased group judgments and extended the findings of a positive relation between stereotype formation and contingency learning. The results support an explanation of biased group judgments by pseudocontingencies; that is, unwarranted inferences from accurately perceived bivariate correlations in complex environments. PMID- 15535775 TI - Inferences about the authentic self: when do actions say more than mental states? AB - Three studies involving 478 undergraduates examined the perceived importance of observable actions versus mental states in revealing the "true self"-the authentic and fundamental nature of a target person. Results suggest that when people have only limited information about a target, they believe that an action is more diagnostic of the individual's true self than the accompanying mental state. When participants have knowledge concerning chronic dispositional tendencies of the target, however, they judge that a chronic mental state is more diagnostic of the true self than a chronic action tendency. Considered together, the findings suggest that people conceptualize the true self as a relatively private entity but nevertheless believe that an action of a little-known person may be particularly informative about that individual. Perceived diagnosticity of the true self was partially mediated by inferences concerning the relative stability of actions versus states but not by inferences of volition. PMID- 15535776 TI - A secure base: responsive support of goal strivings and exploration in adult intimate relationships. AB - A theoretical framework is proposed for examining the interpersonal processes involved in the support of a relationship partner's goal strivings, personal growth, and exploratory behavior, and for examining consequences of receiving either responsive or unresponsive support in this domain. These processes were examined using both observational and experimental methods. In Phase 1, couples were videotaped as they discussed personal goals for the future. In Phase 2, support behavior was experimentally manipulated to examine immediate effects on the recipient. Results indicated that responsive (nonintrusive) support of a relationship partner's goal strivings and explorations have important implications for the recipient's happiness, self-esteem, and perceived likelihood of achieving specific goals. The importance of research examining this type of support is discussed. PMID- 15535777 TI - Put your money where your mouth is! Explaining collective action tendencies through group-based anger and group efficacy. AB - Insights from appraisal theories of emotion are used to integrate elements of theories on collective action. Three experiments with disadvantaged groups systematically manipulated procedural fairness (Study 1), emotional social support (Study 2), and instrumental social support (Study 3) to examine their effects on collective action tendencies through group-based anger and group efficacy. Results of structural equation modeling showed that procedural fairness and emotional social support affected the group-based anger pathway (reflecting emotion-focused coping), whereas instrumental social support affected the group efficacy pathway (reflecting problem-focused coping), constituting 2 distinct pathways to collective action tendencies. Analyses of the means suggest that collective action tendencies become stronger the more fellow group members "put their money where their mouth is." The authors discuss how their dual pathway model integrates and extends elements of current approaches to collective action. PMID- 15535778 TI - How do I love thee? Let me count the Js: implicit egotism and interpersonal attraction. AB - From the perspective of implicit egotism people should gravitate toward others who resemble them because similar others activate people's positive, automatic associations about themselves. Four archival studies and 3 experiments supported this hypothesis. Studies 1-4 showed that people are disproportionately likely to marry others whose first or last names resemble their own. Studies 5-7 provided experimental support for implicit egotism. Participants were more attracted than usual to people (a) whose arbitrary experimental code numbers resembled their own birthday numbers, (b) whose surnames shared letters with their own surnames, and (c) whose jersey number had been paired, subliminally, with their own names. Discussion focuses on implications for implicit egotism, similarity, and interpersonal attraction. PMID- 15535779 TI - Interoceptive sensitivity and self-reports of emotional experience. AB - People differ in the extent to which they emphasize feelings of activation or deactivation in their verbal reports of experienced emotion, termed arousal focus (AF). Two multimethod studies indicate that AF is linked to heightened interoceptive sensitivity (as measured by performance on a heartbeat detection task). People who were more sensitive to their heartbeats emphasized feelings of activation and deactivation when reporting their experiences of emotion over time more than did those who were less sensitive. This relationship was not accounted for by several other variables, including simple language effects. Implications for the role of interoception in experienced emotion and the validity of self reported emotion are discussed. PMID- 15535780 TI - Night and day: are siblings as different in temperament as parents say they are? AB - Twin studies suggest that parent ratings of temperament exaggerate differences between twins. The present study examined whether such contrast effects also operate for nontwin siblings. The activity level (AL) and shyness of 95 nontwin sibling pairs (ages 3 to 8 years) were assessed via parent ratings and objective measures (actigraph and observer ratings). Siblings showed no resemblance in either parent-rated AL or shyness; however, sibling resemblance for actigraph AL and observer-rated shyness was substantial. Thus, parents do contrast their nontwin siblings when rating these 2 temperament dimensions. Moreover, the importance of sibling differences in temperament to the sibling relationship and differential maternal treatment varied across the different measures of AL and shyness, suggesting that parent perceptions may play a role in these associations. PMID- 15535781 TI - A hierarchical analysis of 1,710 English personality-descriptive adjectives. AB - The structure of the English personality lexicon was investigated using self ratings (N = 310) on a set of 1,710 personality-trait adjectives. The 5-factor solution resembled the Big Five structure, but included rotational variants of Agreeableness and Emotional Stability similar to those of other languages. In the 6-factor solution an additional factor, defined by terms such as unpretentious versus sly, resembled an Honesty-Humility factor observed in other languages. The 6-factor solution also produced an especially clear 5th factor, defined by Intellect, Imagination, and Unconventionality content. The hierarchical emergence of factors from 1 to 7 was explored, and the 7-factor solution yielded a Religiosity factor, adding to the diverse array of 7th factors observed in other languages. PMID- 15535782 TI - Trajectories of alcohol and drug use and dependence from adolescence to adulthood: the effects of familial alcoholism and personality. AB - This study describes trajectories of substance use and dependence from adolescence to adulthood. Identified consumption groups include heavy drinking/heavy drug use, moderate drinking/experimental drug use, and light drinking/rare drug use. Dependence groups include alcohol only, drug only, and comorbid groups. The heavy drinking/heavy drug use group was at risk for alcohol and drug dependence and persistent dependence and showed more familial alcoholism, negative emotionality, and low constraint. The moderate drinking/experimental drug use group was at risk for alcohol dependence but not comorbid or persistent dependence and showed less negative emotionality and higher constraint. Familial alcoholism raised risk for alcohol and drug use and dependence in part because children from alcoholic families were more impulsive and lower in agreeableness. PMID- 15535784 TI - Identification of tone duration, line length, and letter position: an experimental approach to timing and working memory deficits in schizophrenia. AB - Patients with schizophrenia display numerous cognitive deficits, including problems in working memory, time estimation, and absolute identification of stimuli. Research in these fields has traditionally been conducted independently. We examined these cognitive processes using tasks that are structurally similar and that yield rich error data. Relative to healthy control participants (n = 20), patients with schizophrenia (n = 20) were impaired on a duration identification task and a probed-recall memory task but not on a line-length identification task. These findings do not support the notion of a global impairment in absolute identification in schizophrenia. However, the authors suggest that some aspect of temporal information processing is indeed disturbed in schizophrenia. PMID- 15535783 TI - Associations in the course of personality disorders and Axis I disorders over time. AB - In this study, the authors examined time-varying associations between schizotypal (STPD), borderline (BPD), avoidant (AVPD), or obsessive-compulsive (OCPD) personality disorders and co-occurring Axis I disorders in 544 adult participants from the Collaborative Longitudinal Personality Disorders Study. The authors tested predictions of specific longitudinal associations derived from a model of crosscutting psychobiological dimensions (L. J. Siever & K. L. Davis, 1991) with participants with the relevant Axis I disorders. The authors assessed participants at baseline and at 6-, 12-, and 24-month follow-up evaluations. BPD showed significant longitudinal associations with major depressive disorder and posttraumatic stress disorder. AVPD was significantly associated with anxiety disorders (specifically social phobia and obsessive-compulsive disorder). Two of the four personality disorders under examination (STPD and OCPD) showed little or no association with Axis I disorders. PMID- 15535785 TI - Psychopathy, risk taking, and attention: a differentiated test of the somatic marker hypothesis. AB - A. R. Damasio's (1994) somatic marker hypothesis relates psychopathy to deficits in the ventromedial prefrontal cortex. Using the gambling task (A. Bechara, A. R. Damasio, H. Damasio, & S. Anderson, 1994), the authors tested this premise and the role of attention as a moderator. Forty-nine male prison inmates were assessed with the Psychopathy Checklist--Revised (R. D. Hare, 1991), the gambling task, and standardized tests on attention-concentration, and intelligence. Results revealed no general relation between psychopathy and gambling task performance. However, psychopathic inmates with low attention scores gambled worse than did the attentive ones. They also had more previous convictions. In nonpsychopathic individuals, attention had no impact. Different processing strategies for psychopathic and nonpsychopathic individuals are proposed to explain these findings. PMID- 15535786 TI - Do college students drink more than their non-college-attending peers? Evidence from a population-based longitudinal female twin study. AB - The association of college attendance with alcohol use and alcohol use disorders was examined in a population-based young adult female twin sample identified from a systematic search of birth records. College-attending women consumed a larger overall volume of alcohol than did their non-college-attending peers, but they were not more likely to be diagnosed with an alcohol use disorder. Significant associations between college attendance and alcohol involvement were probed using 3 different complementary research designs: multivariate cross-sectional analyses, longitudinal analyses of the precollege and college years, and cotwin control analyses of twin pairs discordant for attending college. Although demographic and lifestyle characteristics accounted for most or all of the association between college attendance and alcohol involvement, there was 1 aspect of drinking behavior, occasionally consuming large quantities of alcohol, that remained significantly associated with college attendance even after controlling for these characteristics or for genetic and family background factors. These results are consistent with the conclusion that some aspect of the college experience may be an important environmental risk factor for this pattern of drinking among young adults. PMID- 15535787 TI - Executive functioning and alcohol-related aggression. AB - The primary goal of this investigation was to determine whether executive functioning (EF) would moderate the alcohol-aggression relation. Participants were 310 (152 men and 158 women) healthy social drinkers between 21 and 35 years of age. EF as well as non-EF skills were measured with 13 validated neuropsychological tests. Following the consumption of either an alcoholic or a placebo beverage, participants were tested on a modified version of the Taylor Aggression Paradigm (S. Taylor, 1967), in which mild electric shocks were received from, and administered to, a fictitious opponent. Aggressive behavior was operationalized as the shock intensities administered to the fictitious opponent. EF was negatively related to aggressive behavior for men, regardless of beverage group, even when controlling for non-EF skills. Furthermore, alcohol increased aggression only for men with lower EF scores. Finally, the mere belief that alcohol was consumed suppressed aggression for women but not for men. PMID- 15535788 TI - Context-processing deficits in schizotypal personality disorder. AB - Research suggests that schizotypal personality disorder (SPD) is a part of the spectrum of schizophrenia-related illnesses. This article hypothesizes that a deficit in the representation and maintenance of context is a core cognitive disturbance in schizophrenia and that SPD individuals should demonstrate context processing deficits. To test this hypothesis, the authors administered 3 versions of their AX-CPT task, designed to assess context processing, to 35 healthy controls and 26 individuals with DSM-IV SPD. They also administered working memory and selective attention tasks. SPD individuals displayed context representation deficits similar to those found in schizophrenia but did not show the same additional deficits in context maintenance. Context processing was strongly associated with working memory and selective attention performance in the SPD individuals. PMID- 15535789 TI - Cognitive ability and risk for alcoholism: short-term memory capacity and intelligence moderate personality risk for alcohol problems. AB - This study tested the hypothesis that short-term memory (STM) capacity moderates the effect of social deviance on alcohol problems. Personality, cognitive ability, and alcohol use and abuse were assessed in the adult offspring of alcoholics (FHPs; n = 153) and the adult offspring of nonalcoholics (FHNs; n = 150). The results revealed that STM capacity moderated the effect of social deviance on alcohol problems, independent of intelligence. High social deviance and high-STM participants had fewer alcohol problems than did high social deviance and low-STM participants. Intelligence also moderated the effect of social deviance on alcohol problems in the same way, independent of STM capacity. FHPs had lower IQs, lower verbal ability, and more response perseveration than FHNs. The results suggest that working memory capacity moderates the risk for alcoholism associated with disinhibited traits. PMID- 15535790 TI - Panic disorder and suicide attempt in the National Comorbidity Survey. AB - Clinical and some epidemiological data conflict concerning the likelihood of suicide attempt in individuals with panic disorder (PD). The purpose of this study was to illuminate the panic disorder-suicide attempt association in the National Comorbidity Survey (NCS; R. C. Kessler et al., 1994). Specifically, suicide attempt histories of those 5,872 respondents answering "yes" or "no" to the suicide attempt question were regressed against lifetime diagnostic histories. Lifetime PD history, in the presence of other disorders, was unrelated to elevated risk of suicide attempt and did not account for additional variance. PD respondents who had made a suicide attempt were characterized by Comorbidity. Epidemiological respondents with lifetime histories of PD alone are not at heightened risk for self-reported suicide attempt. PMID- 15535791 TI - Causal attributions about schizophrenia in families in China: expressed emotion and patient relapse. AB - Previous studies have indicated a robust link between relatives' causal attributions and levels of expressed emotion (EE). However, these studies have primarily been conducted in Western cultures. The current study, conducted in China, examined the spontaneous causal attributions made by 54 relatives of schizophrenia patients during the Camberwell Family Interview. Chinese relatives made few controllable and personal attributions overall. Yet as predicted, highly critical and/or hostile EE relatives attributed patients' negative behaviors to more controllable and personal factors. High EE and controllable attributions positively predicted relapse, whereas personal attributions unexpectedly protected against relapse. EE mediated the effect of controllable, but not personal, attributions on relapse. Relatives' use of a particular Chinese characteristic (narrow-mindedness) was integral to the personal dimension's protective effect. PMID- 15535792 TI - Cognitive-neuropsychological function in chronic physical aggression and hyperactivity. AB - Histories of violence and of hyperactivity are both characterized by poor cognitive-neuropsychological function. However, researchers do not know whether these histories combine in additive or interactive ways. The authors tested 303 male young adults from a community sample whose trajectories of teacher-rated physical aggression and motoric hyperactivity from kindergarten to age 15 were well defined. No significant interaction was found. In a 1st model, both histories of problem behavior were independently associated with cognitive neuropsychological function in most domains. In a second model controlling for IQ, General Memory, and test motivation, none of the three Working Memory tests (relevant to executive function) remained associated with physical aggression or hyperactivity. These results support an additive model but no specificity to executive function [corrected]. PMID- 15535793 TI - Evaluating the endophenotype model of ADHD neuropsychological deficit: results for parents and siblings of children with ADHD combined and inattentive subtypes. AB - Neurogenetic models predict neuropsychological weaknesses in the relatives of children with attention-deficit/ hyperactivity disorder (ADHD). The authors examined executive and regulatory measures in 386 relatives (307 parents, 79 siblings) of children with ADHD combined type, ADHD inattentive type, and controls. Predicted deficits were seen on trailmaking (relatives of ADHD combined type only), stop-signal reaction times (relatives of girls only), and response variability (mothers only) but not on naming or output speed. Effects generally held, even with relatives' ADHD status controlled. A neuropsychologically impaired subgroup of children with ADHD had relatives with clear neuropsychological weaknesses. The authors conclude that a neurogenetic model of ADHD etiology is supportable only for a subset of executive functions and that neuropsychological heterogeneity warrants more examination in ADHD. PMID- 15535794 TI - A taxometric analysis of psychopathic personality. AB - Although a common assumption throughout much of the professional and popular literature is that psychopaths are qualitatively different from others, very few studies have examined the latent structure of psychopathy. Whether psychopathy is a discrete category or a continuous dimension may have implications for research on the assessment, etiology, and treatment of this disorder. This study examined the latent structure of psychopathy in a sample of 309 jail and prison inmates. Three taxometric procedures were used to analyze indicators drawn from the Psychopathic Personality Inventory (S. O. Lilienfeld & B. P. Andrews, 1996), a self-report instrument. Consistent with prior studies that used other measures of psychopathy, none of the analyses was consistent with a taxonic solution, suggesting that psychopathic personality may be best understood as existing on a continuum. PMID- 15535795 TI - Externalizing and internalizing subtypes of combat-related PTSD: a replication and extension using the PSY-5 scales. AB - This study replicated and extended prior findings of internalizing and externalizing subtypes of posttraumatic response (M. W. Miller, J. L. Greif, & A. A. Smith, 2003). Cluster analyses of the Minnesota Multiphasic Personality Inventory--2 Personality Psychopathology--Five (MMPI-2 PSY-5; A. R. Harkness, J. L. McNulty, Y. S. Ben-Porath, 1995) profiles obtained from 736 veterans with posttraumatic stress disorder (PTSD) partitioned the sample into a low pathology cluster defined by personality scores in the normal range, an externalizing cluster characterized by low constraint and high negative emotionality, and an internalizing cluster with high negative emotionality and low positive emotionality. Externalizers showed the highest rates of alcohol-related and antisocial personality disorders; internalizers, the highest rates of panic and major depressive disorder. These findings support the development of a personality-based typology of posttraumatic response designed to account for heterogeneity in the expression of PTSD and associated psychopathology. PMID- 15535796 TI - Covariation bias for ambiguous social stimuli in generalized social phobia. AB - The authors investigated whether the negative interpretation bias in generalized social phobia (GSP) reflects and is maintained by illusory correlations. Participants were exposed to descriptions of ambiguous social events, situations involving fear-relevant animals and nature scenes that were randomly paired with negative, positive, or neutral emotional facial expressions. Prior to the experiment, the GSP participants overestimated the contingency social situations negative outcome, whereas the controls judged negative outcomes as least likely. A posteriori, the GSP participants exhibited an illusory correlation specifically between social cues and negative outcomes. During the experiment, only the controls showed distorted outcome predictions for social situations. Hence, illusory correlations--possibly resulting from acquired associations between social cues and negative consequences--may contribute to a negative interpretation bias in GSP. PMID- 15535797 TI - Categorical and dimensional reports of experienced affect to emotion-inducing pictures in depression. AB - Self-reported affect to positive and negative emotional pictures was contrasted in people with major depressive disorder (MDD) and never-depressed control participants (n = 25 in each group). The results revealed significant differences in response to positive images (reduced arousal, less pleasant valence, decreased happiness, increased sadness) in MDD but no clear group differences in response to negative stimuli. Extending earlier findings of reduced responsiveness to positive, but not negative, stimuli in MDD (D. M. Sloan, M. E. Strauss, S. W. Quirk, & M. Satajovik, 1997; D. M. Sloan, M. E. Strauss, & K. L. Wisner, 2001), the data indicate that blunted response to positive stimuli is found when both categorical and dimensional ratings are elicited. Further, the data replicate earlier findings of elevated sadness reports to positive stimuli (J. Rottenberg, K. L. Kasch, J. J. Gross, & I. H. Gotlib, 2002), which may reflect broader difficulties in regulating emotions in MDD. PMID- 15535798 TI - Catalytic mechanism of alpha-retaining glucosyl transfer by Corynebacterium callunae starch phosphorylase: the role of histidine-334 examined through kinetic characterization of site-directed mutants. AB - Purified site-directed mutants of Corynebacterium callunae starch phosphorylase in which His-334 was replaced by an alanine, glutamine or asparagine residue were characterized by steady-state kinetic analysis of enzymic glycosyl transfer to and from phosphate and studies of ligand binding to the active site. Compared with wild-type, the catalytic efficiencies for phosphorolysis of starch at 30 degrees C and pH 7.0 decreased approx. 150- and 50-fold in H334Q (His334-->Gln) and H334N mutants, and that of H334A was unchanged. In the direction of alpha glucan synthesis, selectivity for the reaction with G1P (alpha-D-glucose 1 phosphate) compared with the selectivity for reaction with alpha-D-xylose 1 phosphate decreased from a wild-type value of approximately 20000 to 2600 and 100 in H334N and H334Q respectively. Binding of G1P to the free enzyme was weakened between 10-fold (H334N, H334Q) and 50-fold (H334A) in the mutants, whereas binding to the complex of enzyme and alpha-glucan was not affected. Quenching of fluorescence of the pyridoxal 5'-phosphate cofactor was used to examine interactions of the inhibitor GL (D-gluconic acid 1,5-lactone) with wild-type and mutant enzymes in transient and steady-state experiments. GL binding to the free enzyme and the enzyme-phosphate complex occurred in a single step. The 50-fold higher constant (K(d)) for GL dissociation from H334Q bound to phosphate resulted from an increased off-rate for the ligand in the mutant, compared with wild-type. A log-log correlation of catalytic-centre activity for phosphorolysis of starch with a reciprocal K(d) value established a linear free-energy relationship (slope=1.19+/-0.07; r2=0.991) across the series of wild-type and mutant enzymes. It reveals that GL in combination with phosphate has properties of a transition state analogue and that the His-334 side chain has a role in selectively stabilizing the transition state of the reaction. PMID- 15535800 TI - Determination of in vivo protein synthesis in human palatine tonsil. AB - The palatine tonsils are constantly exposed to ingested or inhaled antigens which, in turn, lead to a permanent activation of tonsillar immune cells, even in a basic physiological state. The aim of the present study was to investigate if the immunological activation of the human palatine tonsil is reflected by a high metabolic activity, as determined by in vivo measurement of protein synthesis. The protein synthesis rate of the tonsil was also compared with that of the circulating T-lymphocytes, the total blood mononuclear cells and the whole population of blood leucocytes. Phenotypic characterization of immune-competent cells in tonsil tissue and blood was performed by flow cytometry. Pinch tonsil biopsies were taken after induction of anaesthesia in healthy adult patients (n=12) scheduled for ear surgery, uvulopalatopharyngoplasty or nose surgery. Protein synthesis was quantitatively determined during a 90-min period by a flooding-dose technique. The in vivo protein synthesis rate in the palatine tonsils was 22.8+/-5.7%/24 h (mean+/-S.D.), whereas protein synthesis in the circulating T-lymphocytes was 10.7+/-3.4%/24 h, in mononuclear cells was 10.8+/ 2.8%/24 h and in leucocytes was 3.2+/-1.2%/24 h. CD3+ lymphocytes were the most abundant cell population in the tonsil. The in vivo protein synthesis rate in human tonsils was higher compared with the circulating immune cells. This high metabolic rate may reflect the permanent immunological activity present in human tonsils, although cell phenotypes and activity markers do not explain the differences. PMID- 15535799 TI - Regulation of human methylthioadenosine phosphorylase gene by the CBF (CCAAT binding factor)/NF-Y (nuclear factor-Y). AB - hMTAP (human 5'-deoxy-5'-methylthioadenosine phosphorylase) is a key enzyme in the methionine salvage pathway and is frequently inactivated in human tumour cells. To understand the mechanism of the transcriptional regulation of the MTAP gene, we have cloned the 1.29 kb fragment of the hMTAP promoter and identified cis-acting regulatory sequences using a luciferase reporter gene assay. Maximal promoter activity was associated with sequences between -446 and -152, where two CCAAT elements were located. Electrophoretic mobility-shift assay reveals binding of specific complexes at both CCAAT motifs within the MTAP promoter, although more prominent bands were associated with the distal motif (-372 to -368). Supershift experiments and chromatin immunoprecipitation assays indicate that both the proximal and distal complexes bind CBF (CCAAT-binding factor; also known as nuclear factor-Y), and that the distal CCAAT motif has increased levels of CBF binding. We have mapped seven different transcriptional start sites between -135 and -58. Our results show that the hMTAP expression is regulated by a CBF and that the distal one of two CCAAT motifs plays a major role in the transcriptional activation of hMTAP gene. PMID- 15535801 TI - Characterization of carnitine and fatty acid metabolism in the long-chain acyl CoA dehydrogenase-deficient mouse. AB - In the present paper, we describe a novel method which enables the analysis of tissue acylcarnitines and carnitine biosynthesis intermediates in the same sample. This method was used to investigate the carnitine and fatty acid metabolism in wild-type and LCAD-/- (long-chain acyl-CoA dehydrogenase-deficient) mice. In agreement with previous results in plasma and bile, we found accumulation of the characteristic C14:1-acylcarnitine in all investigated tissues from LCAD-/- mice. Surprisingly, quantitatively relevant levels of 3 hydroxyacylcarnitines were found to be present in heart, muscle and brain in wild type mice, suggesting that, in these tissues, long-chain 3-hydroxyacyl-CoA dehydrogenase is rate-limiting for mitochondrial beta-oxidation. The 3 hydroxyacylcarnitines were absent in LCAD-/- tissues, indicating that, in this situation, the beta-oxidation flux is limited by the LCAD deficiency. A profound deficiency of acetylcarnitine was observed in LCAD-/- hearts, which most likely corresponds with low cardiac levels of acetyl-CoA. Since there was no carnitine deficiency and only a marginal elevation of potentially cardiotoxic acylcarnitines, we conclude from these data that the cardiomyopathy in the LCAD-/ mouse is caused primarily by a severe energy deficiency in the heart, stressing the important role of LCAD in cardiac fatty acid metabolism in the mouse. PMID- 15535802 TI - Perturbation of actin dynamics induces NF-kappaB activation in myelomonocytic cells through an NADPH oxidase-dependent pathway. AB - Although several reports showed the effect of compounds disrupting microtubules on NF-kappaB (nuclear factor kappaB) activation, nothing is known about agents perturbing actin dynamics. In the present study, we have shown that actin cytoskeleton disruption induced by actin-depolymerizing agents such as cytochalasin D and latrunculin B and actin-polymerizing compounds such as jasplakinolide induced NF-kappaB activation in myelomonocytic cells. The transduction pathway involved the IkappaB (inhibitory kappaB) kinase complex and a degradation of IkappaBalpha. We have shown that NF-kappaB activation in response to the perturbation of actin dynamics required reactive oxygen species, as demonstrated by the effect of antioxidants. Actin cytoskeleton disruption by cytochalasin D induced O2- release from human monocytes, through the activation of the NADPH oxidase, as confirmed by the phosphorylation and by the membrane translocation of p47phox. NF-kappaB activation after actin cytoskeleton disruption could be physiologically relevant during monocyte activation and/or recruitment into injured tissues, where cellular attachment, migration and phagocytosis result in cyclic shifts in cytoskeletal organization and disorganization. PMID- 15535804 TI - Are multi-layer backpropagation networks catastrophically amnesic? AB - Connectionist models with a backpropagation learning rule are known to have a serious problem. Such models exhibit catastrophic interference (or forgetting) with sequential training. Having learned a set of patterns, if the model is trained on another set of patterns, its performance on the first set can dramatically deteriorate very rapidly. The present study reconsiders this issue with four simulations. The model learned arithmetic facts sequentially, but the interference was only modest with random (hence approximately orthogonal) inputs. Essentially the same result was obtained when the inputs are made less orthogonal by adding irrelevant elements. Reducing the number of hidden units did not have major effects. This study suggests that the interference problem has been somewhat overstated. PMID- 15535803 TI - Conformation and concerted dynamics of the integrin-binding site and the C terminal region of echistatin revealed by homonuclear NMR. AB - Echistatin is a potent antagonist of the integrins alpha(v)beta3, alpha5beta1 and alpha(IIb)beta3. Its full inhibitory activity depends on an RGD (Arg-Gly-Asp) motif expressed at the tip of the integrin-binding loop and on its C-terminal tail. Previous NMR structures of echistatin showed a poorly defined integrin recognition sequence and an incomplete C-terminal tail, which left the molecular basis of the functional synergy between the RGD loop and the C-terminal region unresolved. We report a high-resolution structure of echistatin and an analysis of its internal motions by off-resonance ROESY (rotating-frame Overhauser enhancement spectroscopy). The full-length C-terminal polypeptide is visible as a beta-hairpin running parallel to the RGD loop and exposing at the tip residues Pro43, His44 and Lys45. The side chains of the amino acids of the RGD motif have well-defined conformations. The integrin-binding loop displays an overall movement with maximal amplitude of 30 degrees . Internal angular motions in the 100-300 ps timescale indicate increased flexibility for the backbone atoms at the base of the integrin-recognition loop. In addition, backbone atoms of the amino acids Ala23 (flanking the R24GD26 tripeptide) and Asp26 of the integrin-binding motif showed increased angular mobility, suggesting the existence of major and minor hinge effects at the base and the tip, respectively, of the RGD loop. A strong network of NOEs (nuclear Overhauser effects) between residues of the RGD loop and the C-terminal tail indicate concerted motions between these two functional regions. A full-length echistatin-alpha(v)beta3 docking model suggests that echistatin's C-terminal amino acids may contact alpha(v)-subunit residues and provides new insights to delineate structure-function correlations. PMID- 15535805 TI - Residence arrangements and well-being: a study of Norwegian adolescents. AB - The purpose of this study was to assess any differences in psychosocial problems among adolescents living with both parents, or with their mother or their father. Any benefits of living with a same-sex parent compared to a parent of the opposite sex, was also analysed. A total of 1,686 adolescents aged 14-15 years participated from 29 schools in Hordaland county, including schools in downtown Bergen and more rural areas. The findings revealed significantly more psychosocial problems among the adolescents living with one parent compared to both parents. Significant differences were also observed between adolescents living in mother custody compared to father custody, indicating more problems among the latter group. Furthermore, girls living with their father had significantly higher levels of psychological symptoms, compared to boys in father custody. Similarly, boys living with their father were involved in more stealing behavior than girls in father custody. However, residence arrangement accounted for only a limited proportion of the variance in the adolescents' psychosocial problems, indicating large within-group variance and overlap between the different custody groups. PMID- 15535806 TI - Effects of an indoor plant on creative task performance and mood. AB - In this study, we investigated the effect of an indoor plant on task performance and on mood. Three room arrangements were used as independent variables: a room with (1) a plant, or (2) a magazine rack with magazines placed in front of the participants, or (3) a room with neither of these objects. Undergraduate students (M= 35, F= 55) performed a task of associating up to 30 words with each of 20 specified words in a room with one of the three room arrangements. Task performance scores showed that female participants performed better in view of the plant in comparison to the magazine rack (p < 0.05). Moreover, mood was better with the plant or the magazine rack in the room compared to the no object condition (p < 0.05). However, the difference in task performance was highly influenced by the evaluation about the plant or the magazine rack. It is suggested that the compatibility between task demand and the environment is an important factor in facilitating task performances. PMID- 15535807 TI - Climate for work group creativity and innovation: Norwegian validation of the team climate inventory (TCI). AB - The present study assessed the psychometric properties and the validity of the Norwegian translation of the Team Climate Inventory (TCI). The TCI is a measure of climate for innovation within groups at work and is based on the four-factor theory of climate for innovation (West, 1990). Cronbach's alpha revealed satisfactory reliabilities and exploratory factor analysis successfully extracted the four original factors as well as a fifth factor that has also been reported in other studies (N = 195 teams from a wide range of professions). Results from confirmatory factor analysis, using a different sample (N = 106 teams from the Norwegian public postal service), suggested that the five-factor solution had the most parsimonious fit. Criterion validity was explored by correlating TCI scores from 92 post offices and 395 postal distribution teams with customer satisfaction scores. Significant positive relationships were found between three of four TCI scales and customer satisfaction. PMID- 15535808 TI - The effects of meaningful irrelevant speech and road traffic noise on teachers' attention, episodic and semantic memory. AB - The aim of the present experiment was to examine the effects of meaningful irrelevant speech and road traffic noise on attention, episodic and semantic memory, and also to examine whether the noise effects were age-dependent. A total of 96 male and female teachers in the age range of 35-45 and 55-65 years were randomly assigned to a silent or the two noise conditions. Noise effects found in episodic memory were limited to a meaningful text, where cued recall contrary to expectations was equally impaired by the two types of noise. However, meaningful irrelevant speech also deteriorated recognition of the text, whereas road traffic noise caused no decrement. Retrieval from two word fluency tests in semantic memory showed strong effects of noise exposure, one affected by meaningful irrelevant speech and the other by road traffic noise. The results implied that both acoustic variation and the semantic interference could be of importance for noise impairments. The expected age-dependent noise effects did not show up. PMID- 15535809 TI - The effects of noise and gender on children's episodic and semantic memory. AB - The main objectives in the present study were to examine meaningful irrelevant speech and road traffic noise effects on episodic and semantic memory, and to evaluate whether gender differences in memory performance interact with noise. A total of 96 subjects, aged 13-14 years (n = 16 boys and 16 girls in each of three groups), were randomly assigned to a silent or two noise conditions. Noise effects found were restricted to impairments from meaningful irrelevant speech on recognition and cued recall of a text in episodic memory and of word comprehension in semantic memory. The obtained noise effect suggests that the meaning of the speech were processed semantically by the pupils, which reduced their ability to comprehend a text that also involved processing of meaning. Meaningful irrelevant speech was also assumed to cause a poorer access to the knowledge base in semantic memory. Girls outperformed boys in episodic and semantic memory materials, but these differences did not interact with noise. PMID- 15535810 TI - Differentiating in-group favoritism from shared reality in intergroup perception. AB - Two basic factors influence mutual ratings of social groups: in-group favoritism (related to the evaluative aspects of a rating) and the perception of shared reality (related to the descriptive aspects). In two studies, we examine the usefulness of Peabody's (1968) method of separating evaluative and descriptive aspects of rating in intergroup judgments. In Study 1, Latvian and Russian students made different evaluations of both groups, but the same groups agreed on the descriptive ratings. In Study 2, male and female psychology students rated each other from own, in-group, and out-group perspectives. The participants did not show any in-group favoritism in their own ratings, but they expected their fellow students to be in-group biased. The participants agreed on the descriptive ratings of both groups. The results demonstrate that shared reality influences intergroup ratings, despite differences in evaluations. PMID- 15535811 TI - Psychometric properties of the Multidimensional Anxiety Scale for Children (MASC) among Icelandic schoolchildren. AB - A psychometric assessment of the Icelandic version of the Multidimensional Anxiety Scale for Children (MASC) was undertaken among Icelandic schoolchildren, between 10 and 15 years of age. In a first study 625 children between 10 and 15 years of age filled in the MASC. A confirmatory factor analysis indicated that the four-factor structure of the instrument was justified. Further, means and standard deviations of the total scale and the subscales were similar to the original normative data reported by March (1997). In a second study the MASC, the Children Depression Inventory (CDI) and the Revised Children's Manifest Anxiety Scale (RCMAS) were administered to a sample of 249 schoolchildren between 10 and 15 years of age. The results supported the convergent and divergent validity of the MASC. Together the two studies show satisfactory psychometric properties of the Icelandic version of the MASC in a normal population. PMID- 15535812 TI - Self-concept and perception of early mother and father behavior in normal and antisocial adolescents. AB - This study examines the self-concept and perceptions of early parental behavior in a group of 277 normal adolescents with respect to differences in age (12-18 years) and gender. A group of 30 antisocial adolescents of similar age (13-19 years) were included as a comparison group. The study was a cross-sectional questionnaire study and the Swedish translation of the Structural Analysis of Social Behavior (SASB) introject and mother/father questionnaires were used as measures of self- and parental concepts. Multivariate analyses of variance show that the normal adolescent self-concept and perception of early parental behavior are positive and that there are no age or gender differences. The early behavior of the mother is perceived more positively than that of the father. A positive self-concept is related to a positive perception of the parent's early behavior. The antisocial group of adolescents shows a more autonomous and negative self concept and more negative perception of early parental behaviors. This was true especially in the group of antisocial girls. The conclusion from the study is that there is no evidence of a chaotic self-concept or tumultuous changes in the perceptions of early parental behavior during normal adolescence, but that an antisocial way of life might be reflected in a more negative and autonomous self concept and in more negative perceptions of early parental behavior during adolescence. PMID- 15535813 TI - Analysis of the decision-making process leading to appendectomy: a grounded theory study. AB - The aim was to develop a theoretical understanding of the decision-making process leading to appendectomy. A qualitative interview study was performed in the grounded theory tradition using the constant comparative method to analyze data. The study setting was one county hospital and two local hospitals in Sweden, where 11 surgeons and 15 surgical nurses were interviewed. A model was developed which suggests that surgeons' decision making regarding appendectomy is formed by the interplay between their medical assessment of the patient's condition and a set of contextual characteristics. The latter consist of three interacting factors: (1) organizational conditions, (2) the professional actors' individual characteristics and interaction, and (3) the personal characteristics of the patient and his or her family or relatives. In case the outcome of medical assessment is ambiguous, the risk evaluation and final decision will be influenced by an interaction of the contextual characteristics. It was concluded that, compared to existing, rational models of decision making, the model presented identified potentially important contextual characteristics and an outline on when they come into play. PMID- 15535814 TI - The pharmaceutical industry's 'wonder drug'. PMID- 15535815 TI - Leisure-time physical activity among pregnant women in the US. AB - Few population-based data exist that describe leisure-time physical activity among pregnant women. The purpose of this study was to characterise the prevalence of leisure activity and to examine characteristics associated with participation in leisure activity during pregnancy. Using data collected from the year 2000 Behavioral Risk Factor Surveillance System, information on leisure activity was collected during telephone interviews from 1979 pregnant women and 44,657 non-pregnant women 18-44 years of age, representative of the US population. The prevalence of any leisure activity in the past month was 65.6%, 95% confidence interval [95% CI] 62.0, 69.1, among pregnant women and 73.1% [95% CI 72.4, 73.9] among non-pregnant women. The prevalence of recommended activity was also lower among pregnant women than non-pregnant women (15.8% vs. 26.1%). The most common leisure activity for pregnant women was walking, followed by activities such as swimming laps, weight lifting, gardening, and aerobics. Among pregnant women, any leisure activity was significantly higher for those with higher education, younger age, and excellent or very good health than for those with fair or poor health. Pregnant women reported less leisure activity than non pregnant women of the same age. Studies to understand the enablers and barriers to participating in leisure activity during pregnancy are needed. PMID- 15535816 TI - Residential mobility during pregnancy. AB - In epidemiological studies of environmental exposures and adverse pregnancy outcomes, maternal residence at delivery is often used to assign an exposure level, based on routinely collected data. In order to examine the potential for exposure misclassification due to residential mobility, we examined maternal mobility according to changes in residence overall, as well as changes in municipality and county. The potential for mobility to be related to pregnancy outcomes was considered by examining the relationship between mobility and risk factors commonly included in investigations of adverse pregnancy outcomes. Previously collected data were studied from 398 population-based control subjects from a case-control study of stillbirths. We compared demographic, lifestyle, medical, pregnancy and environmental factors of women who moved during pregnancy with those who did not. Bivariable and multivariable log binomial regressions were used to identify risk factors that were associated with mobility during pregnancy. Twelve per cent of subjects moved at least once during their pregnancy. Among women who moved, the majority (62%) moved within the same municipality. In bivariable analyses, we found that low family income, lower maternal age, unmarried status and tobacco use were associated with an increased likelihood of moving during pregnancy, whereas women who used folic acid before conception and who had a higher prepregnancy body mass index (BMI) were less likely to move during pregnancy. In multivariable analyses, only family income, age and prepregnancy BMI were independently predictive of mobility. These results indicate that in studies using maternal residence at delivery to assign environmental exposures, mobility during pregnancy is likely to be prevalent enough to introduce exposure misclassification. The potential for differential exposure misclassification should be considered should any of the risk factors for moving identified by this study also be risk factors for the outcome under study. PMID- 15535817 TI - Descriptive epidemiology of selected congenital heart defects, Hawaii, 1986-1999. AB - Congenital heart defects are the most common type of birth defect and contribute the most to infant mortality due to birth defects. This study examined the relationship between several demographic factors and selected congenital heart defects among the unique multiethnic population in Hawaii during 1986-99, using data from a population-based birth defects registry. Rates were significantly higher in 1993-99 than in 1986-92 for transposition of the great arteries and Ebstein's anomaly, and significantly lower for tetralogy of Fallot. Significantly elevated rates were found with maternal age of > or =35 years for ventricular septal defect, atrial septal defect, endocardial cushion defect, and hypoplastic left heart syndrome. When cases with a known chromosomal abnormality were excluded, elevated rates among the older maternal age group remained for ventricular septal defect, atrial septal defect, and hypoplastic left heart syndrome. Whites had significantly higher rates than one or more of the other racial/ethnic groups for Ebstein's anomaly and coarctation of aorta, and significantly lower rates for tetralogy of Fallot, atrial septal defect, pulmonary valve atresia/stenosis, tricuspid valve atresia/stenosis, and anomalous pulmonary venous return. Significantly higher rates were found among males for transposition of great arteries, aortic valve stenosis, and interrupted aortic arch and, among females, for ventricular septal defect, endocardial cushion defect, and anomalous pulmonary venous return. Some of these differences were consistent with the literature while others were not. PMID- 15535818 TI - Risk factors in term children for visual impairment without a known prenatal or postnatal cause. AB - Risk factors were studied for visual impairment in children without known pre- or postnatal cause, for a decrease of visual acuity. Children born at term 1979-98 and with a visual impairment were identified from the Swedish Register of Visually Impaired Children and data were linked with the Swedish Medical Birth Registry. Maternal characteristics such as maternal age, parity, maternal smoking habits in early pregnancy, maternal education, nationality, and subfertility were studied as well as maternal diagnoses such as pre-eclampsia, prolonged second stage of labour, abruptio placentae, and placenta praevia. Mode of delivery was analysed as well as birthweight, and birthweight in relation to gestational age. Abruptio placentae turned out to be the strongest risk factor (OR = 8.24 [95% CI 5.01, 13.51]). Smoking did not give a statistically significant increased risk. There is an increased risk with breech delivery (OR = 2.01 [95% CI 1.28, 3.17]). Pre-eclampsia was associated with an increased risk (OR = 2.22 [95% CI 1.46, 3.38]). There is also an increase in risk at low birthweight and small-for gestational-age as well as birthweight > 4 kg and large-for-gestational-age. In this study we found that risk factors particularly worth noticing in term children with a presumed perinatal cause of visual impairment are abruptio placentae, pre-eclampsia, excessively low as well as excessively high birthweight, and breech delivery, a fact worth noticing in current discussion on risks, advantages or excessive exploitation of caesarean section. PMID- 15535819 TI - The German study on sleep-disordered breathing in primary school children: epidemiological approach, representativeness of study sample, and preliminary screening results. AB - In order to study the prevalence of habitual snoring (HS), obstructive sleep apnoea (OSA), other aspects of sleep-disordered breathing (SDB), and their determinants, we enrolled a cross-section of children from the population of third graders attending regular primary schools in Hannover, Germany. The recruitment strategy followed a multistage, probability-clustered design. Selected schools were compared with eligible schools regarding the percentage of children with low socio-economic status (SES). The recruitment process was analysed and response proportions calculated. Study subjects were compared with their school enrolment cohort based on anonymous data provided by school authorities. Habitual snoring and OSA were investigated using parental questionnaires and all children underwent nocturnal home pulse oximetry. There was a good overall representativeness of sampled schools with regard to the percentage of children with low SES. In total, 1760 children were contacted and 1144 (65.0%) agreed to participate. The whole process of self-selection of study subjects was not differential by gender. Compared with their school enrolment cohort, study subjects had slightly higher educated parents. The prevalence of HS was estimated to be 10.1% [95% CI 8.3, 11.9%]; and 3.2% [2.2, 4.2%] and 3.9% [2.8, 4.2%] had an abnormally high OSA or SDB score respectively. Based on pulse oximetry, 2.4% [95% CI 1.5, 3.4%] had 3 D(90)and 3 D(C), 1.0% [0.4, 1.6%] had the DI(90) > 0.6, and 3.3% [2.2, 4.4%] had the DI(4) > 3.9 and the DI(C) > 0.4. This study sample appears to be highly representative with regard to all available data from the population, and therefore suitable to study the prevalence of HS and OSA. Our prevalence estimates are in the range of other studies. PMID- 15535820 TI - Association of early-onset otitis media in infants and exposure to household mould. AB - Otitis media is one of the most common infections of early childhood. Children who first experience acute otitis media at an early age (before 6 months) are at increased risk for recurrent otitis media. This prospective study investigated exposure to measured levels of airborne household mould and the risk of early otitis media in the first 6 months of life among a cohort of infants at high risk for asthma. Between September 1996 and December 1998, women were invited to participate if they had at least one other child with physician-diagnosed asthma. Mothers were given a standardised questionnaire within 4 months of their infant's birth. Airborne mould samples were also taken at this time, and culturable fungi were categorised into four levels according to the report of the Commission of European Communities: 0 (undetectable), 1-499 colony forming units (CFU)/m(3) (low), 500-999 CFU/m(3) (medium), > or =1000 CFU/m(3) (high). Infant respiratory symptoms were collected during quarterly telephone interviews at 6, 9 and 12 months of age. Of the 806 children in the study, 27.8% experienced otitis media before six months of age. Household levels of Penicillium and Cladosporium were modestly associated with the number of otitis media episodes (P = 0.056 and 0.081 respectively). After controlling for potential confounders, Penicillium and Cladosporium were not associated with early otitis media. High levels of 'other' mould (defined as total spore count minus counts for Penicillium, Cladosporium, and yeast) were associated with early otitis media (OR 3.49; 95% CI [1.38, 8.79]). We also found associations between day-care outside of the home and birth during the summer or fall season with early otitis media. This study is suggestive of a relationship between otitis media and mould that warrants further study. PMID- 15535821 TI - Methods for identifying pregnancy-associated deaths: population-based data from Finland 1987-2000. AB - To find maternal and pregnancy-related deaths, it is important that all pregnancy associated deaths are identified. This article examines the effect of data linkages between national health care registers and complete death certificate data on pregnancy-associated deaths. All deaths among women of reproductive age (15-49 years) in Finland during the period 1987-2000 (n = 15 823) were identified from the Cause-of-Death Register and linked to the Medical Birth Register (n = 865 988 births), the Register on Induced Abortions (n = 156 789 induced abortions), and the Hospital Discharge Register (n = 118 490 spontaneous abortions) to determine whether women had been pregnant within 1 year before death. The death certificates of the 419 women thus identified were reviewed to find whether the pregnancy or its termination was coded or mentioned. In total, 405 deaths (96.7%) were identified in registers other than the Cause-of-Death Register. Without data linkages, 73% of all pregnancy-associated deaths would have been missed; the percentage after induced and spontaneous abortions was even higher. Data linkages to national health care registers provide better information on maternal deaths and pregnancy-associated deaths than death certificates alone. If possible, pregnancies not ending in a live birth should be included in the data linkages. PMID- 15535822 TI - Artificial neural networks as statistical tools in epidemiological studies: analysis of risk factors for early infant wheeze. AB - Artificial neural networks (ANNs) are being used increasingly for the prediction of clinical outcomes and classification of disease phenotypes. A lack of understanding of the statistical principles underlying ANNs has led to widespread misuse of these tools in the biomedical arena. In this paper, the authors compare the performance of ANNs with that of conventional linear logistic regression models in an epidemiological study of infant wheeze. Data on the putative risk factors for infant wheeze have been obtained from a sample of 7318 infants taking part in the Avon Longitudinal Study of Parents and Children (ALSPAC). The data were analysed using logistic regression models and ANNs, and performance based on misclassification rates of a validation data set were compared. Misclassification rates in the training data set decreased as the complexity of the ANN increased: h = 0: 17.9%; h = 2: 16.2%; h = 5: 14.9%, and h = 10: 9.2%. However, the more complex models did not generalise well to new data sets drawn from the same population: validation data set misclassification rates: h = 0: 17.9%; h = 2: 19.6%; h = 5: 20.2% and h = 10: 22.9%. There is no evidence from this study that ANNs outperform conventional methods of analysing epidemiological data. Increasing the complexity of the models serves only to overfit the model to the data. It is important that a validation or test data set is used to assess the performance of highly complex ANNs to avoid overfitting. PMID- 15535823 TI - Multiple sudden infant deaths--coincidence or beyond coincidence? PMID- 15535827 TI - Acoustic stiffness and change in plug cartilage over time after autologous osteochondral grafting: correlation between ultrasound signal intensity and histological score in a rabbit model. AB - We investigated quantitative changes over time in ultrasound signal intensity (an index of stiffness), signal duration (an index of surface irregularity), and interval between signals (an index of thickness) of plug cartilage in an animal model of autologous osteochondral grafting. A full-thickness osteochondral plug was surgically removed and replaced in male Japanese white rabbits (n = 22). Specimens obtained at day 0 and weeks 2, 4, 8, 12 and 24 postoperatively were assessed using an ultrasound system and by macroscopic and histological evaluation (modified Mankin's score). Histology revealed that the plug sank until 2 weeks postoperatively, and that newly formed cartilage-like tissue covered the plug, but at 24 weeks the tissue detached. The plug itself survived well throughout the period of observation. Although the signal intensity at the plug site was same as that in the sham operated contralateral knee at day 0, from 2 to 24 weeks postoperatively it was less than that in the sham knee. At 8 weeks, this difference was significant (P < 0.05). Modified Mankin's score revealed early degenerative changes at the site, but macroscopic examination did not. Signal intensity correlated significantly with score (both at day 0 and at the five postoperative time points [P < 0.05, r = -0.91] and as a whole [P < 0.05, r = 0.36]). Signal intensity also significantly correlated with the individual subscores for 'cartilage structure' (P < 0.05, r = -0.32) and 'cartilage cells' (P < 0.05, r = -0.30) from the modified Mankin's score, but not significantly with subscores for 'staining' and 'tidemark'. Signal duration correlated significantly with total score (as a whole [P < 0.05, r = 0.34]), but not significantly with the score for cartilage structure (P = 0.0557, r = 0.29). The interval between signals reflected well the actual thickness of the plug site. The significant relationships between ultrasound signal intensity and scores suggest that early degenerative changes in plug cartilage and cartilage-like tissue, especially in the superficial layer, are detectable by high-frequency ultrasound assessment. PMID- 15535828 TI - Percentile benchmarks in patients with rheumatoid arthritis: Health Assessment Questionnaire as a quality indicator (QI). AB - Physicians are in need of a simple objective, standardized tool to compare their patients with rheumatoid arthritis, as a group and individually, with national standards. The Disability Index of the Health Assessment Questionnaire (HAQ-DI) is a simple, robust tool that can fulfill these needs. However, use of this tool as a quality indicator (QI) is hampered by the unavailability of national reference values or benchmarks based on large, multicentric, heterogenous longitudinal patient cohorts. We utilized the 20-year longitudinal prospective data from 11 data banks of Arthritis Rheumatism and Aging Medical Information to calculate reference values for HAQ-DI. Overall, 6436 patients with rheumatoid arthritis were longitudinally followed for 32,324 person-years over the 20 years from 1981 to 2000. There were 64,647 HAQ-DI measurements, with an average of 19 measurements per person. Overall, 75% of patients were women and 89% were Caucasian; the median baseline age was 58.4 years and the median baseline HAQ-DI was 1.13. Few patients were treated with biologics. The HAQ-DI values had a Gaussian distribution except for the approximately 10% of observations showing no disability. Percentile benchmarks allow disability outcomes to be compared and contrasted between different patient populations. Reference values for the HAQ DI, presented here numerically and graphically, can be used in clinical practice as a QI measure to track functional disability outcomes and to measure response to therapy, and by arthritis patients in self-management programs. PMID- 15535829 TI - Expression analysis of three isoforms of hyaluronan synthase and hyaluronidase in the synovium of knees in osteoarthritis and rheumatoid arthritis by quantitative real-time reverse transcriptase polymerase chain reaction. AB - Hyaluronan is a major molecule in joint fluid and plays a crucial role in joint motion and the maintenance of joint homeostasis. The concentration and average molecular weight of hyaluronan in the joint fluids are reduced in osteoarthritis and rheumatoid arthritis. To elucidate the underlying mechanism, we analyzed the message expression of three isoforms of hyaluronan synthase and hyaluronidase from knee synovium, using real-time reverse transcriptase polymerase chain reaction. Synovia were obtained from 17 patients with osteoarthritis, 14 patients with rheumatoid arthritis, and 20 healthy control donors. The message expression of hyaluronan synthase-1 and -2 in the synovium of both types of arthritis was significantly less than in the control synovium, whereas that of hyaluronidase-2 in the synovium of both arthritides was significantly greater than in the control synovium. The decreased expression of the messages for hyaluronan synthase-1 and 2 and/or the increased expression of the message for hyaluronidase-2 may be reflected in the reduced concentration and decreased average molecular weight of hyaluronan in the joint fluids of patients with osteoarthritis and rheumatoid arthritis. PMID- 15535830 TI - Use of HLA-B27 tetramers to identify low-frequency antigen-specific T cells in Chlamydia-triggered reactive arthritis. AB - Reports of the use of HLA-B27/peptide tetrameric complexes to study peptide specific CD8+ T cells in HLA-B27+-related diseases are rare. To establish HLA-B27 tetramers we first compared the function of HLA-B27 tetramers with HLA-A2 tetramers by using viral epitopes. HLA-B27 and HLA-A2 tetramers loaded with immunodominant peptides from Epstein-Barr virus were generated with comparable yields and both molecules detected antigen-specific CD8+ T cells. The application of HLA-B27 tetramers in HLA-B27-related diseases was performed with nine recently described Chlamydia-derived peptides in synovial fluid and peripheral blood, to examine the CD8+ T cell response against Chlamydia trachomatis antigens in nine patients with Chlamydia-triggered reactive arthritis (Ct-ReA). Four of six HLA B27+ Ct-ReA patients had specific synovial T cell binding to at least one HLA B27/Chlamydia peptide tetramer. The HLA-B27/Chlamydia peptide 195 tetramer bound to synovial T cells from three of six patients and HLA-B27/Chlamydia peptide 133 tetramer to synovial T cells from two patients. However, the frequency of these cells was low (0.02-0.09%). Moreover, we demonstrate two methods to generate HLA B27-restricted T cell lines. First, HLA-B27 tetramers and magnetic beads were used to sort antigen-specific CD8+ T cells. Second, Chlamydia-infected dendritic cells were used to stimulate CD8+ T cells ex vivo. Highly pure CD8 T cell lines could be generated ex vivo by magnetic sorting by using HLA-B27 tetramers loaded with an EBV peptide. The frequency of Chlamydia-specific, HLA-B27 tetramer binding CD8+ T cells could be increased by stimulating CD8+ T cells ex vivo with Chlamydia-infected dendritic cells. We conclude that HLA-B27 tetramers are a useful tool for the detection and expansion of HLA-B27-restricted CD8+ T cells. T cells specific for one or more of three Chlamydia-derived peptides were found at low frequency in synovial fluid from HLA-B27+ patients with Ct-ReA. These cells can be expanded ex vivo, suggesting that they are immunologically functional. PMID- 15535831 TI - Infliximab therapy in rheumatoid arthritis and ankylosing spondylitis-induced specific antinuclear and antiphospholipid autoantibodies without autoimmune clinical manifestations: a two-year prospective study. AB - Treatment of rheumatoid arthritis (RA) with infliximab (Remicade) has been associated with the induction of antinuclear autoantibodies (ANA) and anti-double stranded DNA (anti-dsDNA) autoantibodies. In the present study we investigated the humoral immune response induced by infliximab against organ-specific or non organ-specific antigens not only in RA patients but also in patients with ankylosing spondylitis (AS) during a two-year followup. The association between the presence of autoantibodies and clinical manifestations was then examined. The occurrence of the various autoantibodies was analyzed in 24 RA and 15 AS patients all treated with infliximab and in 30 RA patients receiving methotrexate but not infliximab, using the appropriate methods of detection. Infliximab led to a significant induction of ANA and anti-dsDNA autoantibodies in 86.7% and 57% of RA patients and in 85% and 31% of AS patients, respectively. The incidence of antiphospholipid (aPL) autoantibodies was significantly higher in both RA patients (21%) and AS patients (27%) than in the control group. Most anti-dsDNA and aPL autoantibodies were of IgM isotype and were not associated with infusion side effects, lupus-like manifestations or infectious disease. No other autoantibodies were shown to be induced by the treatment. Our results confirmed the occurrence of ANA and anti-dsDNA autoantibodies and demonstrated that the induction of ANA, anti-dsDNA and aPL autoantibodies is related to infliximab treatment in both RA and AS, with no significant relationship to clinical manifestations. PMID- 15535832 TI - Collagen type II (CII)-specific antibodies induce arthritis in the absence of T or B cells but the arthritis progression is enhanced by CII-reactive T cells. AB - Antibodies against type II collagen (anti-CII) are arthritogenic and have a crucial role in the initiation of collagen-induced arthritis. Here, we have determined the dependence of T and B cells in collagen-antibody-induced arthritis (CAIA) during different phases of arthritis. Mice deficient for B and/or T cells were susceptible to the CAIA, showing that the antibodies induce arthritis even in the absence of an adaptive immune system. To determine whether CII-reactive T cells could have a role in enhancing arthritis development at the effector level of arthritis pathogenesis, we established a T cell line reactive with CII. This T cell line was oligoclonal and responded to different post-translational forms of the major CII epitope at position 260-270 bound to the Aq class II molecule. Importantly, it cross-reacted with the mouse peptide although it is bound with lower affinity to the Aq molecule than the corresponding rat peptide. The T cell line could not induce clinical arthritis per se in Aq-expressing mice even if these mice expressed the major heterologous CII epitope in cartilage, as in the transgenic MMC (mutated mouse collagen) mouse. However, a combined treatment with anti-CII monoclonal antibodies and CII-reactive T cells enhanced the progression of severe arthritis. PMID- 15535833 TI - Intrathecal levels of matrix metalloproteinases in systemic lupus erythematosus with central nervous system engagement. AB - Symptoms originating from the central nervous system (CNS) occur frequently in patients with systemic lupus erythematosus (SLE), and CNS involvement in lupus is associated with increased morbidity and mortality. We recently showed that neurones and astrocytes are continuously damaged during the course of CNS lupus. The matrix metalloproteinases (MMPs) are a group of tissue degrading enzymes that may be involved in this ongoing brain destruction. The aim of this study was to examine endogenous levels of free, enzymatically active MMP-2 and MMP-9 in cerebrospinal fluid from patients with SLE. A total of 123 patients with SLE were evaluated clinically, with magnetic resonance imaging of brain and cerebrospinal fluid (CSF) analyses. Levels of free MMP-2 and MMP-9 were determined in CSF using an enzymatic activity assay. CSF samples from another 22 cerebrally healthy individuals were used as a control. Intrathecal MMP-9 levels were significantly increased in patients with neuropsychiatric SLE as compared with SLE patients without CNS involvement (P < 0.05) and healthy control individuals (P = 0.0012). Interestingly, significant correlations between MMP-9 and intrathecal levels of neuronal and glial degradation products were noted, indicating ongoing intrathecal degeneration in the brains of lupus patients expressing MMP-9. In addition, intrathecal levels of IL-6 and IL-8--two cytokines that are known to upregulate MMP-9--both exhibited significant correlation with MMP-9 levels in CSF (P < 0.0001), suggesting a potential MMP-9 activation pathway. Our findings suggest that proinflammatory cytokine induced MMP-9 production leads to brain damage in patients with CNS lupus. PMID- 15535834 TI - Analysis of HLA DR, HLA DQ, C4A, FcgammaRIIa, FcgammaRIIIa, MBL, and IL-1Ra allelic variants in Caucasian systemic lupus erythematosus patients suggests an effect of the combined FcgammaRIIa R/R and IL-1Ra 2/2 genotypes on disease susceptibility. AB - Dysfunction in various parts of immune defence, such as immune response, immune complex clearance, and inflammation, has an impact on pathogenesis in systemic lupus erythematosus (SLE). We hypothesised that combinations of common variants of genes involved in these immune functions are associated with susceptibility to SLE. The following variants were analysed: HLA DR3, HLA DQ2, C4AQ0, Fcgamma receptor IIa (FcgammaRIIa) genotype R/R, Fcgamma receptor IIIa (FcRgammaIIIa) genotype F/F, mannan-binding lectin (MBL) genotype conferring a low serum concentration of MBL (MBL-low), and interleukin-1 receptor antagonist (IL-1Ra) genotype 2/2. Polymorphisms were analysed in 143 Caucasian patients with SLE and 200 healthy controls. HLA DR3 in SLE patients was in 90% part of the haplotype HLA DR3-DQ2-C4AQ0, which was strongly associated with SLE (odds ratio [OR] 2.8, 95% CI 1.7-4.5). Analysis of combinations of gene variants revealed that the strong association with SLE for HLA DR3-DQ2-C4AQ0 remained after combination with FcgammaRIIa R/R, FcgammaRIIIa F/F, and MBL-low (OR>2). Furthermore, the combination of the FcgammaRIIa R/R and IL-1Ra 2/2 genotypes yielded a strong correlation with SLE (OR 11.8, 95% CI 1.5-95.4). This study demonstrates that certain combinations of gene variants may increase susceptibility to SLE, suggesting this approach for future studies. It also confirms earlier findings regarding the HLA DR3-DQ2-C4AQ0 haplotype. PMID- 15535835 TI - Resistance to IL-10 inhibition of interferon gamma production and expression of suppressor of cytokine signaling 1 in CD4+ T cells from patients with rheumatoid arthritis. AB - IL-10 has been shown to block the antigen-specific T-cell cytokine response by inhibiting the CD28 signaling pathway. We found that peripheral blood CD4+ T cells from patients with active rheumatoid arthritis (RA) were able to produce greater amounts of interferon gamma after CD3 and CD28 costimulation in the presence of 1 ng/ml IL-10 than were normal control CD4+ T cells, although their surface expression of the type 1 IL-10 receptor was increased. The phosphorylation of signal transducer and activator of transcription 3 was sustained in both blood and synovial tissue CD4+ T cells of RA, but it was not augmented by the presence of 1 ng/ml IL-10. Sera from RA patients induced signal transducer and activator of transcription 3 phosphorylation in normal CD4+ T cells, which was mostly abolished by neutralizing anti-IL-6 antibody. Preincubation of normal CD4+ T cells with IL-6 reduced IL-10-mediated inhibition of interferon gamma production. Blood CD4+ T cells from RA patients contained higher levels of suppressor of cytokine signaling 1 but lower levels of suppressor of cytokine signaling 3 mRNA compared with control CD4+ T cells, as determined by real-time PCR. These results indicate that RA CD4+ T cells become resistant to the immunosuppressive effect of IL-10 before migration into synovial tissue, and this impaired IL-10 signaling may be associated with sustained signal transducer and activator of transcription 3 activation and suppressor of cytokine signaling 1 induction. PMID- 15535836 TI - Ethics of randomised controlled trials--not yet time to give up on equipoise. AB - In this commentary on Fries and Krishnan's argument that 'design bias' undermines the status of equipoise as the ethical justification for randomised controlled trials, it is argued that their argument is analogous to Bayesian arguments for the use of informative priors in trial design, but that this does not undermine the importance of equipoise. In particular, mismatches between the outcomes of interest to industrial sponsors of research and outcomes of interest to patients and clinicians ensure that in many cases industry-sponsored trials can fail to reflect the reasonable equipoise of working clinicians. PMID- 15535837 TI - Biology of recently discovered cytokines: interleukin-17--a unique inflammatory cytokine with roles in bone biology and arthritis. AB - IL-17 and its receptor are founding members of an emerging family of cytokines and receptors with many unique characteristics. IL-17 is produced primarily by T cells, particularly those of the memory compartment. In contrast, IL-17 receptor is ubiquitously expressed, making nearly all cells potential targets of IL-17. Although it has only limited homology to other cytokines, IL-17 exhibits proinflammatory properties similar to those of tumor necrosis factor-alpha, particularly with respect to induction of other inflammatory effectors. In addition, IL-17 synergizes potently with other cytokines, placing it in the center of the inflammatory network. Strikingly, IL-17 has been associated with several bone pathologies, most notably rheumatoid arthritis. PMID- 15535840 TI - What can chronic arthritis pain teach about developing new analgesic drugs? AB - Chronic pain remains an important public health need with greater impact on the US economy than most other chronic conditions. Current pain management is largely limited to opioids and non-steroidal anti-inflammatory drugs, indicating a gap in the translation of new knowledge to the development of improved pain treatments. Strategies suggested include the re-evaluation of current drug screening methods, a recognition that molecular-genetic events occurring acutely contribute to the development of pain chronicity, the validation of analgesic targets in the intended patient population, consideration of the unique genetic profile that varies between individuals, and the introduction of individual response measures to improve the capture of outcomes in clinical trials. PMID- 15535838 TI - The tandem CCCH zinc finger protein tristetraprolin and its relevance to cytokine mRNA turnover and arthritis. AB - Tristetraprolin (TTP) is the best-studied member of a small family of three proteins in humans that is characterized by a tandem CCCH zinc finger (TZF) domain with highly conserved sequences and spacing. Although initially discovered as a gene that could be induced rapidly and transiently by the stimulation of fibroblasts with growth factors and mitogens, it is now known that TTP can bind to AU-rich elements in mRNA, leading to the removal of the poly(A) tail from that mRNA and increased rates of mRNA turnover. This activity was discovered after TTP deficient mice were created and found to have a systemic inflammatory syndrome with severe polyarticular arthritis and autoimmunity, as well as medullary and extramedullary myeloid hyperplasia. The syndrome seemed to be due predominantly to excess circulating tumor necrosis factor-alpha (TNF-alpha), resulting from the increased stability of the TNF-alpha mRNA and subsequent higher rates of secretion of the cytokine. The myeloid hyperplasia might be due in part to increased stability of granulocyte-macrophage colony-stimulating factor (GM-CSF). This review highlights briefly the characteristics of the TTP-deficiency syndrome in mice and its possible genetic modifiers, as well as recent data on the characteristics of the TTP-binding site in the TNF-alpha and GM-CSF mRNAs. Recent structural data on the characteristics of the complex between RNA and one of the TTP-related proteins are reviewed, and used to model the TTP-RNA binding complex. We review the current knowledge of TTP sequence variants in humans and discuss the possible contributions of the TTP-related proteins in mouse physiology and in human monocytes. The TTP pathway of TNF-alpha and GM-CSF mRNA degradation is a possible novel target for anti-TNF-alpha therapies for rheumatoid arthritis, and also for other conditions proven to respond to anti-TNF-alpha therapy. PMID- 15535839 TI - Oxidation in rheumatoid arthritis. AB - Oxygen metabolism has an important role in the pathogenesis of rheumatoid arthritis. Reactive oxygen species (ROS) produced in the course of cellular oxidative phosphorylation, and by activated phagocytic cells during oxidative bursts, exceed the physiological buffering capacity and result in oxidative stress. The excessive production of ROS can damage protein, lipids, nucleic acids, and matrix components. They also serve as important intracellular signaling molecules that amplify the synovial inflammatory-proliferative response. Repetitive cycles of hypoxia and reoxygenation associated with changes in synovial perfusion are postulated to activate hypoxia-inducible factor-1alpha and nuclear factor-kappaB, two key transcription factors that are regulated by changes in cellular oxygenation and cytokine stimulation, and that in turn orchestrate the expression of a spectrum of genes critical to the persistence of synovitis. An understanding of the complex interactions involved in these pathways might allow the development of novel therapeutic strategies for rheumatoid arthritis. PMID- 15535841 TI - Autoantibodies in normals--the value of predicting rheumatoid arthritis. PMID- 15535842 TI - Role of Notch signaling in cell-fate determination of human mammary stem/progenitor cells. AB - INTRODUCTION: Notch signaling has been implicated in the regulation of cell-fate decisions such as self-renewal of adult stem cells and differentiation of progenitor cells along a particular lineage. Moreover, depending on the cellular and developmental context, the Notch pathway acts as a regulator of cell survival and cell proliferation. Abnormal expression of Notch receptors has been found in different types of epithelial metaplastic lesions and neoplastic lesions, suggesting that Notch may act as a proto-oncogene. The vertebrate Notch1 and Notch4 homologs are involved in normal development of the mammary gland, and mutated forms of these genes are associated with development of mouse mammary tumors. METHODS: In order to determine the role of Notch signaling in mammary cell-fate determination, we have utilized a newly described in vitro system in which mammary stem/progenitor cells can be cultured in suspension as nonadherent 'mammospheres'. Notch signaling was activated using exogenous ligands, or was inhibited using previously characterized Notch signaling antagonists. RESULTS: Utilizing this system, we demonstrate that Notch signaling can act on mammary stem cells to promote self-renewal and on early progenitor cells to promote their proliferation, as demonstrated by a 10-fold increase in secondary mammosphere formation upon addition of a Notch-activating DSL peptide. In addition to acting on stem cells, Notch signaling is also able to act on multipotent progenitor cells, facilitating myoepithelial lineage-specific commitment and proliferation. Stimulation of this pathway also promotes branching morphogenesis in three dimensional Matrigel cultures. These effects are completely inhibited by a Notch4 blocking antibody or a gamma secretase inhibitor that blocks Notch processing. In contrast to the effects of Notch signaling on mammary stem/progenitor cells, modulation of this pathway has no discernable effect on fully committed, differentiated, mammary epithelial cells. CONCLUSION: These studies suggest that Notch signaling plays a critical role in normal human mammary development by acting on both stem cells and progenitor cells, affecting self-renewal and lineage-specific differentiation. Based on these findings we propose that abnormal Notch signaling may contribute to mammary carcinogenesis by deregulating the self-renewal of normal mammary stem cells. PMID- 15535843 TI - The Na+-H+ exchanger-1 induces cytoskeletal changes involving reciprocal RhoA and Rac1 signaling, resulting in motility and invasion in MDA-MB-435 cells. AB - INTRODUCTION: An increasing body of evidence shows that the tumour microenvironment is essential in driving neoplastic progression. The low serum component of this microenvironment stimulates motility/invasion in human breast cancer cells via activation of the Na+-H+ exchanger (NHE) isoform 1, but the signal transduction systems that underlie this process are still poorly understood. We undertook the present study to elucidate the role and pattern of regulation by the Rho GTPases of this serum deprivation-dependent activation of both NHE1 and subsequent invasive characteristics, such as pseudopodia and invadiopodia protrusion, directed cell motility and penetration of normal tissues. METHODS: The present study was performed in a well characterized human mammary epithelial cell line representing late stage metastatic progression, MDA MB-435. The activity of RhoA and Rac1 was modified using their dominant negative and constitutively active mutants and the activity of NHE1, cell motility/invasion, F-actin content and cell shape were measured. RESULTS: We show for the first time that serum deprivation induces NHE1-dependent morphological and cytoskeletal changes in metastatic cells via a reciprocal interaction of RhoA and Rac1, resulting in increased chemotaxis and invasion. Deprivation changed cell shape by reducing the amount of F-actin and inducing the formation of leading edge pseudopodia. Serum deprivation inhibited RhoA activity and stimulated Rac1 activity. Rac1 and RhoA were antagonistic regulators of both basal and stimulated tumour cell NHE1 activity. The regulation of NHE1 activity by RhoA and Rac1 in both conditions was mediated by an alteration in intracellular proton affinity of the exchanger. Interestingly, the role of each of these G-proteins was reversed during serum deprivation; basal NHE1 activity was regulated positively by RhoA and negatively by Rac1, whereas RhoA negatively and Rac1 positively directed the stimulation of NHE1 during serum deprivation. Importantly, the same pattern of RhoA and Rac1 regulation found for NHE1 activity was observed in both basal and serum deprivation dependent increases in motility, invasion and actin cytoskeletal organization. CONCLUSION: Our findings suggest that the reported antagonistic roles of RhoA and Rac1 in cell motility/invasion and cytoskeletal organization may be due, in part, to their concerted action on NHE1 activity as a convergence point. PMID- 15535844 TI - Frequency of CHEK2 mutations in a population based, case-control study of breast cancer in young women. AB - INTRODUCTION: The cell-cycle checkpoint kinase (CHEK)2 protein truncating mutation 1100delC has been associated with increased risk for breast or prostate cancer. Multiple studies have found an elevated frequency of the 1100delC variant in specific stratifications of breast cancer patients with a family history of the disease, including BRCA1/BRCA2 negative families and families with a history of bilateral disease or male breast cancer. However, the 1100delC mutation has only been investigated in a few population-based studies and none from North America. METHODS: We report here on the frequency of three CHEK2 variants that alter protein function--1100delC, R145W, and I175T--in 506 cases and 459 controls from a population based, case-control study of breast cancer conducted in young women from western Washington. RESULTS: There was a suggestive enrichment in the 1100delC variant in the cases (1.2%) as compared with the controls (0.4%), but this was based on small numbers of carriers and the differences were not statistically significant. The 1100delC variant was more frequent in cases with a first-degree family history of breast cancer (4.3%; P = 0.02) and slightly enriched in cases with a family history of ovarian cancer (4.4%; P = 0.09). CONCLUSION: The CHEK2 variants are rare in the western Washington population and, based on accumulated evidence across studies, are unlikely to be major breast cancer susceptibility genes. Thus, screening for the 1100delC variant may have limited usefulness in breast cancer prevention programs in the USA. PMID- 15535845 TI - The progesterone receptor Val660-->Leu polymorphism and breast cancer risk. AB - BACKGROUND: Recent evidence suggests a role for progesterone in breast cancer development and tumorigenesis. Progesterone exerts its effect on target cells by interacting with its receptor; thus, genetic variations, which might cause alterations in the biological function in the progesterone receptor (PGR), can potentially contribute to an individual's susceptibility to breast cancer. It has been reported that the PROGINS allele, which is in complete linkage disequilibrium with a missense substitution in exon 4 (G/T, valine-->leucine, at codon 660), is associated with a decreased risk for breast cancer. METHODS: Using a nested case-control study design within the Nurses' Health Study cohort, we genotyped 1252 cases and 1660 matched controls with the use of the Taqman assay. RESULTS: We did not observe any association of breast cancer risk with carrying the G/T (Val660-->Leu) polymorphism (odds ratio 1.10, 95% confidence interval 0.93-1.30). In addition, we did not observe an interaction between this allele and menopausal status and family history of breast cancer as reported previously. CONCLUSION: Overall, our study does not support an association between the Val660 ->Leu PROGINS polymorphism and breast cancer risk. PMID- 15535846 TI - Over-expression of lysophosphatidic acid receptor-2 in human invasive ductal carcinoma. AB - INTRODUCTION: Lysophosphatidic acid (LPA) is a bioactive phospholipid with diverse effects on various cells. It interacts with at least three G-protein coupled transmembrane receptors, namely LPA1, LPA2 and LPA3, whose expression in various tumours has not been fully characterized. In the present study we characterized the expression profile of LPA receptors in human breast cancer tissue and assessed the possible roles of each receptor. METHODS: The relative expression levels of each receptor's mRNA against beta-actin mRNA was examined in surgically resected invasive ductal carcinomas and normal gland tissue using real time RT-PCR. LPA2 expression was also examined immunohistochemically using a rat anti-LPA2 monoclonal antibody. RESULTS: In 25 cases normal and cancer tissue contained LPA1 mRNA at similar levels, whereas the expression level of LPA2 mRNA was significantly increased in cancer tissue as compared with its normal counterpart (3479.0 +/- 426.6 versus 1287.3 +/- 466.8; P < 0.05). LPA3 was weakly expressed in both cancer and normal gland tissue. In 48 (57%) out of 84 cases, enhanced expression of LPA2 protein was confirmed in carcinoma cells as compared with normal mammary epithelium by immunohistochemistry. Over-expression of LPA2 was detected in 17 (45%) out of 38 premenopausal women, as compared with 31 (67%) out of 46 postmenopausal women, and the difference was statistically significant (P < 0.05). CONCLUSION: These findings suggest that upregulation of LPA2 may play a role in carcinogenesis, particularly in postmenopausal breast cancer. PMID- 15535847 TI - Genetic polymorphism in the manganese superoxide dismutase gene, antioxidant intake, and breast cancer risk: results from the Shanghai Breast Cancer Study. AB - INTRODUCTION: It has been suggested that oxidative stress and mitochondrial DNA damage play important roles in breast cancer carcinogenesis. Manganese superoxide dismutase (MnSOD) is a major enzyme that is responsible for the detoxification of reactive oxygen species in the mitochondria. A T --> C substitution in the MnSOD gene results in a Val --> Ala change at the -9 position of the mitochondrial targeting sequence (Val-9Ala), which alters the protein secondary structure and thus affects transport of MnSOD into the mitochondria. METHODS: We evaluated this genetic polymorphism in association with breast cancer risk using data from the Shanghai Breast Cancer Study, a population-based case-control study conducted in urban Shanghai from 1996 to 1998. The MnSOD Val-9Ala polymorphism was examined in 1125 breast cancer cases and 1197 age-frequency-matched control individual. RESULTS: Breast cancer risk was slightly elevated in women with Ala/Ala genotype (odds ratio [OR] 1.3, 95% confidence interval [CI] 0.7-2.3), particularly among premenopausal women (OR 1.8, 95% CI 0.9-3.7), as compared with those with Val/Val genotype. The increased risk with the Ala/Ala genotype was stronger among premenopausal women with a higher body mass index (OR 2.5, 95% CI 0.9-7.0) and more years of menstruation (OR 2.6, 95% CI 0.8-8.0). The risk among premenopausal women was further increased twofold to threefold among those with a low intake of fruits, vegetables, vitamin supplements, selenium, or antioxidant vitamins, including carotenes and vitamins A, C, and E. However, the frequency of the Ala allele was low (14%) in the study population, and most of the ORs provided above were not statistically significant. CONCLUSION: The present study provides some evidence that genetic polymorphism in the MnSOD gene may be associated with increased risk of breast cancer among Chinese women with high levels of oxidative stress or low intake of antioxidants. Studies with a larger sample size are needed to confirm the findings. PMID- 15535848 TI - Birthweight, parental age, birth order and breast cancer risk in African-American and white women: a population-based case-control study. AB - INTRODUCTION: Much recent work has focused on hypotheses that very early life exposures influence adult cancer risk. For breast cancer it has been hypothesized that high in utero estrogen exposure may increase risk. METHODS: We used data from the Carolina Breast Cancer Study, a population-based case-control study of incident breast cancer in North Carolina, to examine associations for three possible surrogates of high prenatal estrogen exposure: weight at birth, maternal age, and birth order. We also examined paternal age. Birthweight analyses were conducted for white and African-American women born in North Carolina on or after 1949 (196 cases, 167 controls). Maternal age was analyzed for US born participants younger than 49 years of age (280 cases, 236 controls). RESULTS: There was a weak inverse association between birthweight in the highest tertile and breast cancer overall (odds ratio [OR] 0.7, 95% confidence interval [CI] 0.4 1.2), although associations differed by race (OR 0.5, 95% CI 0.2-1.0, and OR 1.0, 95% CI 0.5-2.1 for African-American and white women, respectively). For maternal age there was an approximately threefold increase in risk in women whose mothers were older than 22 years of age, relative to 19-22 years of age, when the women were born. After adjustment for maternal age, older paternal age increased risk in the oldest and youngest age categories (relative to 23-27 years of age at the woman's birth: OR 1.6, 95% CI 0.8-3.1 for age 15-22 years; OR 1.2, 95% CI 0.7-2.2 for age 28-34 years; and OR 1.5, 95% CI 0.7-3.2 for age 35-56 years). There was no association with older paternal age for white women alone. After adjustment for maternal age (265 cases, 224 controls), a birth order of fifth or higher relative to first had an inverse association with breast cancer for women younger than 49 years old (OR 0.6, 95% CI 0.3-1.3). CONCLUSION: Although the CIs are wide, these results lend support to the possibility that the prenatal period is important for subsequent breast cancer risk, but they do not support the estrogen hypothesis as a unifying theory for the influence of this period. PMID- 15535849 TI - Proteotypic classification of spontaneous and transgenic mammary neoplasms. AB - INTRODUCTION: Mammary tumors in mice are categorized by using morphologic and architectural criteria. Immunolabeling for terminal differentiation markers was compared among a variety of mouse mammary neoplasms because expression of terminal differentiation markers, and especially of keratins, provides important information on the origin of neoplastic cells and their degree of differentiation. METHODS: Expression patterns for terminal differentiation markers were used to characterize tumor types and to study tumor progression in transgenic mouse models of mammary neoplasia (mice overexpressing Neu (Erbb2), Hras, Myc, Notch4, SV40-TAg, Tgfa, and Wnt1), in spontaneous mammary carcinomas, and in mammary neoplasms associated with infection by the mouse mammary tumor virus (MMTV). RESULTS: On the basis of the expression of terminal differentiation markers, three types of neoplasm were identified: first, simple carcinomas composed exclusively of cells with a luminal phenotype are characteristic of neoplasms arising in mice transgenic for Neu, Hras, Myc, Notch4, and SV40-TAg; second, 'complex carcinomas' displaying luminal and myoepithelial differentiation are characteristic of type P tumors arising in mice transgenic for Wnt1, neoplasms arising in mice infected by the MMTV, and spontaneous adenosquamous carcinomas; and third, 'carcinomas with epithelial to mesenchymal transition (EMT)' are a characteristic feature of tumor progression in Hras-, Myc-, and SV40 TAg-induced mammary neoplasms and PL/J and SJL/J mouse strains, and display de novo expression of myoepithelial and mesenchymal cell markers. In sharp contrast, EMT was not detected in papillary adenocarcinomas arising in BALB/cJ mice, spontaneous adenoacanthomas, neoplasms associated with MMTV-infection, or in neoplasms arising in mice transgenic for Neu and Wnt1. CONCLUSIONS: Immunohistochemical profiles of complex neoplasms are consistent with a stem cell origin, whereas simple carcinomas might originate from a cell committed to the luminal lineage. In addition, these results suggest that the initiating oncogenic events determine the morphologic features associated with cancer progression because EMT is observed only in certain types of neoplasm. PMID- 15535850 TI - Ratios of involved nodes in early breast cancer. AB - INTRODUCTION: The number of lymph nodes found to be involved in an axillary dissection is among the most powerful prognostic factors in breast cancer, but it is confounded by the number of lymph nodes that have been examined. We investigate an idea that has surfaced recently in the literature (since 1999), namely that the proportion of node-positive lymph nodes (or a function thereof) is a much better predictor of survival than the number of excised and node positive lymph nodes, alone or together. METHODS: The data were abstracted from 83,686 cases registered in the Surveillance, Epidemiology, and End Results (SEER) program of women diagnosed with nonmetastatic T1-T2 primary breast carcinoma between 1988 and 1997, in whom axillary node dissection was performed. The end point was death from breast cancer. Cox models based on different expressions of nodal involvement were compared using the Nagelkerke R2 index (R2N). Ratios were modeled as percentage and as log odds of involved nodes. Log odds were estimated in a way that avoids singularities (zero values) by using the empirical logistic transform. RESULTS: In node-negative cases both the number of nodes excised and the log odds were significant, with hazard ratios of 0.991 (95% confidence interval 0.986-0.997) and 1.150 (1.058-1.249), respectively, but without improving R2N. In node-positive cases the hazard ratios were 1.003-1.088 for the number of involved nodes, 0.966-1.005 for the number of excised nodes, 1.015 1.017 for the percentage, and 1.344-1.381 for the log odds. R2N improved from 0.067 (no nodal covariate) to 0.102 (models based on counts only) and to 0.108 (models based on ratios). DISCUSSION: Ratios are simple optimal predictors, in that they provide at least the same prognostic value as the more traditional staging based on counting of involved nodes, without replacing them with a needlessly complicated alternative. They can be viewed as a per patient standardization in which the number of involved nodes is standardized to the number of nodes excised. In an extension to the study, ratios were validated in a comparison with categorized staging measures using blinded data from the San Jose Monterey cancer registry. A ratio based prognostic index was also derived. It improved the Nottingham Prognostic Index without compromising on simplicity. PMID- 15535851 TI - Menopausal status dependence of the timing of breast cancer recurrence after surgical removal of the primary tumour. AB - INTRODUCTION: Information on the metastasis process in breast cancer patients undergoing primary tumour removal may be extracted from an analysis of the timing of clinical recurrence. METHODS: The hazard rate for local-regional and/or distant recurrence as the first event during the first 4 years after surgery was studied in 1173 patients undergoing mastectomy alone as primary treatment for operable breast cancer. Subset analyses were performed according to tumour size, axillary nodal status and menopausal status. RESULTS: A sharp two-peaked hazard function was observed for node-positive pre-menopausal patients, whereas results from node-positive post-menopausal women always displayed a single broad peak. The first narrow peak among pre-menopausal women showed a very steep rise to a maximum about 8-10 months after mastectomy. The second peak was considerably broader, reaching its maximum at 28-30 months. Post-menopausal patients displayed a wide, nearly symmetrical peak with maximum risk at about 18-20 months. Peaks displayed increasing height with increasing axillary lymph node involvement. No multi-peaked pattern was evident for either pre-menopausal or post-menopausal node-negative patients; however, this finding should be considered cautiously because of the limited number of events. Tumour size influenced recurrence risk but not its timing. Findings resulting from the different subsets of patients were remarkably coherent and each observed peak maintained the same position on the time axis in all analysed subsets. CONCLUSIONS: The risk of early recurrence for node positive patients is dependent on menopausal status. The amount of axillary nodal involvement and the tumour size modulate the risk value at any given time. For pre-menopausal node-positive patients, the abrupt increase of the first narrow peak of the recurrence risk suggests a triggering event that synchronises early risk. We suggest that this event is the surgical removal of the primary tumour. The later, broader, more symmetrical risk peaks indicate that some features of the corresponding metastatic development may present stochastic traits. A metastasis development model incorporating tumour dormancy in specific micro-metastatic phases, stochastic transitions between them and sudden acceleration of the metastatic process by surgery can explain these risk dynamics. PMID- 15535852 TI - The changing global patterns of female breast cancer incidence and mortality. AB - One in ten of all new cancers diagnosed worldwide each year is a cancer of the female breast, and it is the most common cancer in women in both developing and developed areas. It is also the principal cause of death from cancer among women globally. We review the descriptive epidemiology of the disease, focusing on some of the key elements of the geographical and temporal variations in incidence and mortality in each world region. The observations are discussed in the context of the numerous aetiological factors, as well as the impact of screening and advances in treatment and disease management in high-resource settings. PMID- 15535853 TI - The origins of estrogen receptor alpha-positive and estrogen receptor alpha negative human breast cancer. AB - Current hormonal therapies have benefited millions of patients with breast cancer. Their success, however, is often temporary and limited to a subset of patients whose tumors express estrogen receptor alpha (ER). The therapies are entirely ineffective in ER-negative disease. Recent studies suggest that there are many biological pathways and alterations involved in determining whether ER is expressed and how it is regulated during breast cancer evolution. Improving hormonal therapies, in addition to perfecting current strategies, will also target these newly discovered pathways and alterations, and others yet to be found. The present commentary will briefly highlight a few important observations and unanswered questions regarding ER status and growth regulation during breast cancer evolution, which hopefully will help to stimulate new thinking and progress in this important area of medial research. PMID- 15535855 TI - An early peak of relapse after surgery for breast cancer. AB - There is great interest among oncologists concerning what we might learn by examining the pattern of relapse after breast cancer surgery. What you see depends upon how hard you look. Up to now, investigators have examined the hazard ratio for relapse every 6-12 months. In a research paper, published in this issue of Breast Cancer Research, the Milan group have looked at the hazard ratio every three months and have found, for the first time, a distinct, very early peak of relapse in a group of premenopausal, node-positive patients not given chemotherapy or hormone therapy. What is now needed is for other groups to repeat this observation and, if found, to examine the characteristics of the tumours producing this phenomenon in order to develop hypotheses about its cause and possible treatments. PMID- 15535854 TI - Metabolic inactivation of estrogens in breast tissue by UDP glucuronosyltransferase enzymes: an overview. AB - The breast tissue is the site of major metabolic conversions of estradiol (E2) mediated by specific cytochromes P450 hydroxylations and methylation by catechol O-methytransferase. In addition to E2 itself, recent findings highlight the significance of 4-hydroxylated estrogen metabolites as chemical mediators and their link to breast cancer development and progression, whereas, in opposition, 2-methoxylated estrogens appear to be protective. Recent data also indicate that breast tissue possesses enzymatic machinery to inactivate and eliminate E2 and its oxidized and methoxylated metabolites through conjugation catalyzed by UDP glucuronosyltransferases (UGTs), which involves the covalent addition of glucuronic acid. In opposition to other metabolic pathways of estrogen, the UGT mediated process leads to the formation of glucuronides that are devoid of biologic activity and are readily excreted from the tissue into the circulation. This review addresses the most recent findings on the identification of UGT enzymes that are responsible for the glucuronidation of E2 and its metabolites, and evidence regarding their potential role in breast cancer. PMID- 15535856 TI - Bone marrow micrometastases and circulating tumor cells: current aspects and future perspectives. AB - Early tumor cell dissemination at the single-cell level can be revealed in patients with breast cancer by using sensitive immunocytochemical and molecular assays. Recent clinical studies involving more than 4000 breast cancer patients demonstrated that the presence of disseminated tumor cells in bone marrow at primary diagnosis is an independent prognostic factor. In addition, various assays for the detection of circulating tumor cells in the peripheral blood have recently been developed and some studies also suggest a potential clinical relevance of this measure. These findings provide the basis for the potential use of disseminated tumor cells in bone marrow or blood as markers for the early assessment of therapeutic response in prospective clinical trials. PMID- 15535857 TI - New targets for therapy in breast cancer: farnesyltransferase inhibitors. AB - Current systemic therapies for breast cancer are often limited by their nonspecific mechanism of action, unwanted toxicities on normal tissues, and short term efficacy due to the emergence of drug resistance. However, identification of the molecular abnormalities in cancer, in particular the key proteins involved in abnormal cell growth, has resulted in development of various signal transduction inhibitor drugs as new treatment strategies against the disease. Protein farnesyltransferase inhibitors (FTIs) were originally designed to target the Ras signal transduction pathway, although it is now clear that several other intracellular proteins are dependent on post-translational farnesylation for their function. Preclinical data revealed that although FTIs inhibit the growth of ras-transformed cells, they are also potent inhibitors of a wide range of cancer cell lines that contain wild-type ras, including breast cancer cells. Additive or synergistic effects were observed when FTIs were combined with cytotoxic agents (in particular the taxanes) or endocrine therapies (tamoxifen). Phase I trials with FTIs have explored different schedules for prolonged administration, and dose-limiting toxicities included myelosuppression, gastrointestinal toxicity and neuropathy. Clinical efficacy against breast cancer was seen for the FTI tipifarnib in a phase II study. Based on promising preclinical data that suggest synergy with taxanes or endocrine therapy, combination clinical studies are now in progress to determine whether FTIs can add further to the efficacy of conventional breast cancer therapies. PMID- 15535858 TI - Endocrinology and hormone therapy in breast cancer: aromatase inhibitors versus antioestrogens. AB - Endocrine therapies act by either blocking or downregulating the oestrogen receptor or by reducing oestrogen concentrations around and within the cancer cell. In postmenopausal women, oestrogen suppression is achieved by inhibition of the enzyme aromatase by aromatase inhibitors (AIs). Modern AIs (anastrozole, letrozole and exemestane) are more potent than earlier ones and suppress oestradiol levels in plasma to virtually undetectable concentrations. Recent comparisons of AIs with the most widely used oestrogen receptor blocking drug tamoxifen indicate that, in general, AIs result in increased response rates and greater durations of response. Here, we summarize data supporting the difference between the two types of treatment and attempt to account for the underlying mechanisms that favour AIs. PMID- 15535859 TI - American Society for Clinical Oncology 40th annual meeting, New Orleans, LA, 5-8 June 2004: breast cancer highlights. PMID- 15535860 TI - Location, location, location: regulation of breast cancer progression by the microenvironment. PMID- 15535861 TI - Transcriptome analysis of haploid male gametophyte development in Arabidopsis. AB - BACKGROUND: The haploid male gametophyte generation of flowering plants consists of two- or three-celled pollen grains. This functional specialization is thought to be a key factor in the evolutionary success of flowering plants. Moreover, pollen ontogeny is also an attractive model in which to dissect cellular networks that control cell growth, asymmetric cell division and cellular differentiation. Our objective, and an essential step towards the detailed understanding of these processes, was to comprehensively define the male haploid transcriptome throughout development. RESULTS: We have developed staged spore isolation procedures for Arabidopsis and used Affymetrix ATH1 genome arrays to identify a total of 13,977 male gametophyte-expressed mRNAs, 9.7% of which were male gametophyte-specific. The transition from bicellular to tricellular pollen was accompanied by a decline in the number of diverse mRNA species and an increase in the proportion of male gametophyte-specific transcripts. Expression profiles of regulatory proteins and distinct clusters of coexpressed genes were identified that could correspond to components of gametophytic regulatory networks. Moreover, integration of transcriptome and experimental data revealed the early synthesis of translation factors and their requirement to support pollen tube growth. CONCLUSIONS: The progression from proliferating microspores to terminally differentiated pollen is characterized by large-scale repression of early program genes and the activation of a unique late gene-expression program in maturing pollen. These data provide a quantum increase in knowledge concerning gametophytic transcription and lay the foundations for new genomic-led studies of the regulatory networks and cellular functions that operate to specify male gametophyte development. PMID- 15535862 TI - Spatial patterns of transcriptional activity in the chromosome of Escherichia coli. AB - BACKGROUND: Although genes on the chromosome are organized in a fixed order, the spatial correlations in transcription have not been systematically evaluated. We used a combination of genomic and signal processing techniques to investigate the properties of transcription in the genome of Escherichia coli K12 as a function of the position of genes on the chromosome. RESULTS: Spectral analysis of transcriptional series revealed the existence of statistically significant patterns in the spatial series of transcriptional activity. These patterns could be classified into three categories: short-range, of up to 16 kilobases (kb); medium-range, over 100-125 kb; and long-range, over 600-800 kb. We show that the significant similarities in gene activities extend beyond the length of an operon and that local patterns of coexpression are dependent on DNA supercoiling. Unlike short-range patterns, the formation of medium and long-range transcriptional patterns does not strictly depend on the level of DNA supercoiling. The long range patterns appear to correlate with the patterns of distribution of DNA gyrase on the bacterial chromosome. CONCLUSIONS: Localization of structural components in the transcriptional signal revealed an asymmetry in the distribution of transcriptional patterns along the bacterial chromosome. The demonstration that spatial patterns of transcription could be modulated pharmacologically and genetically, along with the identification of molecular correlates of transcriptional patterns, offer for the first time strong evidence of physiologically determined higher-order organization of transcription in the bacterial chromosome. PMID- 15535863 TI - Genomic transcriptional response to loss of chromosomal supercoiling in Escherichia coli. AB - BACKGROUND: The chromosome of Escherichia coli is maintained in a negatively supercoiled state, and supercoiling levels are affected by growth phase and a variety of environmental stimuli. In turn, supercoiling influences local DNA structure and can affect gene expression. We used microarrays representing nearly the entire genome of Escherichia coli MG1655 to examine the dynamics of chromosome structure. RESULTS: We measured the transcriptional response to a loss of supercoiling caused either by genetic impairment of a topoisomerase or addition of specific topoisomerase inhibitors during log-phase growth and identified genes whose changes are statistically significant. Transcription of 7% of the genome (306 genes) was rapidly and reproducibly affected by changes in the level of supercoiling; the expression of 106 genes increased upon chromosome relaxation and the expression of 200 decreased. These changes are most likely to be direct effects, as the kinetics of their induction or repression closely follow the kinetics of DNA relaxation in the cells. Unexpectedly, the genes induced by relaxation have a significantly enriched AT content in both upstream and coding regions. CONCLUSIONS: The 306 supercoiling-sensitive genes are functionally diverse and widely dispersed throughout the chromosome. We propose that supercoiling acts as a second messenger that transmits information about the environment to many regulatory networks in the cell. PMID- 15535864 TI - Phylogenomic evidence supports past endosymbiosis, intracellular and horizontal gene transfer in Cryptosporidium parvum. AB - BACKGROUND: The apicomplexan parasite Cryptosporidium parvum is an emerging pathogen capable of causing illness in humans and other animals and death in immunocompromised individuals. No effective treatment is available and the genome sequence has recently been completed. This parasite differs from other apicomplexans in its lack of a plastid organelle, the apicoplast. Gene transfer, either intracellular from an endosymbiont/donor organelle or horizontal from another organism, can provide evidence of a previous endosymbiotic relationship and/or alter the genetic repertoire of the host organism. Given the importance of gene transfers in eukaryotic evolution and the potential implications for chemotherapy, it is important to identify the complement of transferred genes in Cryptosporidium. RESULTS: We have identified 31 genes of likely plastid/endosymbiont (n = 7) or prokaryotic (n = 24) origin using a phylogenomic approach. The findings support the hypothesis that Cryptosporidium evolved from a plastid-containing lineage and subsequently lost its apicoplast during evolution. Expression analyses of candidate genes of algal and eubacterial origin show that these genes are expressed and developmentally regulated during the life cycle of C. parvum. CONCLUSIONS: Cryptosporidium is the recipient of a large number of transferred genes, many of which are not shared by other apicomplexan parasites. Genes transferred from distant phylogenetic sources, such as eubacteria, may be potential targets for therapeutic drugs owing to their phylogenetic distance or the lack of homologs in the host. The successful integration and expression of the transferred genes in this genome has changed the genetic and metabolic repertoire of the parasite. PMID- 15535865 TI - Assembly and characterization of heterochromatin and euchromatin on human artificial chromosomes. AB - BACKGROUND: Human centromere regions are characterized by the presence of alpha satellite DNA, replication late in S phase and a heterochromatic appearance. Recent models propose that the centromere is organized into conserved chromatin domains in which chromatin containing CenH3 (centromere-specific H3 variant) at the functional centromere (kinetochore) forms within regions of heterochromatin. To address these models, we assayed formation of heterochromatin and euchromatin on de novo human artificial chromosomes containing alpha-satellite DNA. We also examined the relationship between chromatin composition and replication timing of artificial chromosomes. RESULTS: Heterochromatin factors (histone H3 lysine 9 methylation and HP1alpha) were enriched on artificial chromosomes estimated to be larger than 3 Mb in size but depleted on those smaller than 3 Mb. All artificial chromosomes assembled markers of euchromatin (histone H3 lysine 4 methylation), which may partly reflect marker-gene expression. Replication timing studies revealed that the replication timing of artificial chromosomes was heterogeneous. Heterochromatin-depleted artificial chromosomes replicated in early S phase whereas heterochromatin-enriched artificial chromosomes replicated in mid to late S phase. CONCLUSIONS: Centromere regions on human artificial chromosomes and host chromosomes have similar amounts of CenH3 but exhibit highly varying degrees of heterochromatin, suggesting that only a small amount of heterochromatin may be required for centromere function. The formation of euchromatin on all artificial chromosomes demonstrates that they can provide a chromosome context suitable for gene expression. The earlier replication of the heterochromatin-depleted artificial chromosomes suggests that replication late in S phase is not a requirement for centromere function. PMID- 15535866 TI - Reconstruction of regulatory and metabolic pathways in metal-reducing delta proteobacteria. AB - BACKGROUND: Relatively little is known about the genetic basis for the unique physiology of metal-reducing genera in the delta subgroup of the proteobacteria. The recent availability of complete finished or draft-quality genome sequences for seven representatives allowed us to investigate the genetic and regulatory factors in a number of key pathways involved in the biosynthesis of building blocks and cofactors, metal-ion homeostasis, stress response, and energy metabolism using a combination of regulatory sequence detection and analysis of genomic context. RESULTS: In the genomes of delta-proteobacteria, we identified candidate binding sites for four regulators of known specificity (BirA, CooA, HrcA, sigma-32), four types of metabolite-binding riboswitches (RFN-, THI-, B12 elements and S-box), and new binding sites for the FUR, ModE, NikR, PerR, and ZUR transcription factors, as well as for the previously uncharacterized factors HcpR and LysX. After reconstruction of the corresponding metabolic pathways and regulatory interactions, we identified possible functions for a large number of previously uncharacterized genes covering a wide range of cellular functions. CONCLUSIONS: Phylogenetically diverse delta-proteobacteria appear to have homologous regulatory components. This study for the first time demonstrates the adaptability of the comparative genomic approach to de novo reconstruction of a regulatory network in a poorly studied taxonomic group of bacteria. Recent efforts in large-scale functional genomic characterization of Desulfovibrio species will provide a unique opportunity to test and expand our predictions. PMID- 15535867 TI - Genome-wide patterns of carbon and nitrogen regulation of gene expression validate the combined carbon and nitrogen (CN)-signaling hypothesis in plants. AB - BACKGROUND: Carbon and nitrogen are two signals that influence plant growth and development. It is known that carbon- and nitrogen-signaling pathways influence one another to affect gene expression, but little is known about which genes are regulated by interactions between carbon and nitrogen signaling or the mechanisms by which the different pathways interact. RESULTS: Microarray analysis was used to study global changes in mRNA levels due to carbon and nitrogen in Arabidopsis thaliana. An informatic analysis using InterAct Class enabled us to classify genes on the basis of their responses to carbon or nitrogen treatments. This analysis provides in vivo evidence supporting the hypothesis that plants have a carbon/nitrogen (CN)-sensing/regulatory mechanism, as we have identified over 300 genes whose response to combined CN treatment is different from that expected from expression values due to carbon and nitrogen treatments separately. Metabolism, energy and protein synthesis were found to be significantly affected by interactions between carbon and nitrogen signaling. Identified putative cis acting regulatory elements involved in mediating CN-responsive gene expression suggest multiple mechanisms for CN responsiveness. One mechanism invokes the existence of a single CN-responsive cis element, while another invokes the existence of cis elements that promote nitrogen-responsive gene expression only when present in combination with a carbon-responsive cis element. CONCLUSION: This study has allowed us to identify genes and processes regulated by interactions between carbon and nitrogen signaling and take a first step in uncovering how carbon- and nitrogen-signaling pathways interact to regulate transcription. PMID- 15535868 TI - Sparse graphical Gaussian modeling of the isoprenoid gene network in Arabidopsis thaliana. AB - We present a novel graphical Gaussian modeling approach for reverse engineering of genetic regulatory networks with many genes and few observations. When applying our approach to infer a gene network for isoprenoid biosynthesis in Arabidopsis thaliana, we detect modules of closely connected genes and candidate genes for possible cross-talk between the isoprenoid pathways. Genes of downstream pathways also fit well into the network. We evaluate our approach in a simulation study and using the yeast galactose network. PMID- 15535869 TI - How biologically relevant are interaction-based modules in protein networks? AB - By applying a graph-based algorithm to yeast protein-interaction networks we have extracted modular structures and show that they can be validated using information from the phylogenetic conservation of the network components. We show that the module cores, the parts with the highest intramodular connectivity, are biologically relevant components of the networks. These constituents correlate only weakly with other levels of organization. We also discuss how such structures could be used for finding targets for antimicrobial drugs. PMID- 15535870 TI - Consensus clustering and functional interpretation of gene-expression data. AB - Microarray analysis using clustering algorithms can suffer from lack of inter method consistency in assigning related gene-expression profiles to clusters. Obtaining a consensus set of clusters from a number of clustering methods should improve confidence in gene-expression analysis. Here we introduce consensus clustering, which provides such an advantage. When coupled with a statistically based gene functional analysis, our method allowed the identification of novel genes regulated by NFkappaB and the unfolded protein response in certain B-cell lymphomas. PMID- 15535871 TI - Integrating transcriptional controls for plant cell expansion. AB - The plant hormones auxin and brassinosteroid promote cell expansion by regulating gene expression. In addition to independent transcriptional responses generated by the two signals, recent microarray analyses indicate that auxin and brassinosteroid also coordinate the expression of a set of shared target genes. PMID- 15535872 TI - Human molecular chronotyping in sight? AB - Recent research on mouse models has taken us closer to deciphering the molecular clock mechanism that defines an individual's 'body time'. How feasible will it be to create a molecular timetable that allows determination of individual body time from tissue harvested at a single time point? PMID- 15535873 TI - Uncovering the complexities of Kaposi's sarcoma through genome-wide expression analysis. AB - Gene-expression profiling of endothelial cells infected with Kaposi's sarcoma associated herpesvirus has led to a greater understanding of the histogenesis of Kaposi's sarcoma and cellular reprogramming events that occur as a result of viral infection and that may play important roles in viral pathogenesis. PMID- 15535874 TI - The 3-hydroxy-3-methylglutaryl coenzyme-A (HMG-CoA) reductases. AB - The enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase catalyzes the conversion of HMG-CoA to mevalonate, a four-electron oxidoreduction that is the rate-limiting step in the synthesis of cholesterol and other isoprenoids. The enzyme is found in eukaryotes and prokaryotes; and phylogenetic analysis has revealed two classes of HMG-CoA reductase, the Class I enzymes of eukaryotes and some archaea and the Class II enzymes of eubacteria and certain other archaea. Three-dimensional structures of the catalytic domain of HMG-CoA reductases from humans and from the bacterium Pseudomonas mevalonii, in conjunction with site directed mutagenesis studies, have revealed details of the mechanism of catalysis. The reaction catalyzed by human HMG-CoA reductase is a target for anti hypercholesterolemic drugs (statins), which are intended to lower cholesterol levels in serum. Eukaryotic forms of the enzyme are anchored to the endoplasmic reticulum, whereas the prokaryotic enzymes are soluble. Probably because of its critical role in cellular cholesterol homeostasis, mammalian HMG-CoA reductase is extensively regulated at the transcriptional, translational, and post translational levels. PMID- 15535875 TI - The dosage-compensation complex in flies and humans. PMID- 15535876 TI - High society (of nematologists). PMID- 15535877 TI - From genomes to systems. PMID- 15535878 TI - Genetic diversity of Plasmodium vivax isolates from Azerbaijan. AB - BACKGROUND: Plasmodium vivax, although causing a less serious disease than Plasmodium falciparum, is the most widespread of the four human malarial species. Further to the recent recrudescence of P. vivax cases in the Newly Independent States (NIS) of central Asia, a survey on the genetic diversity and dissemination in Azerbaijan was undertaken. Azerbaijan is at the crossroads of Asia and, as such, could see a rise in the number of cases, although an effective malaria control programme has been established in the country. METHODS: Thirty-six P. vivax isolates from Central Azerbaijan were characterized by analysing the genetic polymorphism of the circumsporozoite protein (CSP) and the merozoite surface protein 1 (MSP-1) genes, using PCR amplifications and amplicons sequencing. RESULTS: Analysis of CSP sequences showed that all the processed isolates belong to the VK 210 type, with variations in the alternation of alanine residue (A) or aspartic acid residue (D) in the repeat motif GDRA(A/D)GQPA along the sequence. As far as MSP-1 genotyping is concerned, it was found that the majority of isolates analysed belong to Belem and Sal I types. Five recombinant isolates were also identified. Combined analysis with the two genetic markers allowed the identification of 19 plasmodial sub-types. CONCLUSION: The results obtained in the present study indicate that there are several P. vivax clones circulating in Azerbaijan and, consequently, a careful malaria surveillance could be of paramount importance to identify, at early stage, the occurrence of possible P. vivax malaria outbreaks. PMID- 15535880 TI - Random allocation software for parallel group randomized trials. AB - BACKGROUND: Typically, randomization software should allow users to exert control over the different aspects of randomization including block design, provision of unique identifiers and control over the format and type of program output. While some of these characteristics have been addressed by available software, none of them have all of these capabilities integrated into one package. The main objective of the Random Allocation Software project was to enhance the user's control over different aspects of randomization in parallel group trials, including output type and format, structure and ordering of generated unique identifiers and enabling users to specify group names for more than two groups. RESULTS: The program has different settings for: simple and blocked randomizations; length, format and ordering of generated unique identifiers; type and format of program output; and saving sessions for future use. A formatted random list generated by this program can be used directly (without further formatting) by the coordinator of the research team to prepare and encode different drugs or instruments necessary for the parallel group trial. CONCLUSIONS: Random Allocation Software enables users to control different attributes of the random allocation sequence and produce qualified lists for parallel group trials. PMID- 15535879 TI - Effect of dietary palm olein oil on oxidative stress associated with ischemic reperfusion injury in isolated rat heart. AB - BACKGROUND: Palm olein oil (PO), obtained from refining of palm oil is rich in monounsaturated fatty acid and antioxidant vitamins and is widely used as oil in diet in many parts of the world including India. Palm oil has been reported to have beneficial effects in oxidative stress associated with hypertension and arterial thrombosis. Oxidative stress plays a major role in the etiopathology of myocardial ischemic-reperfusion injury (IRI) which is a common sequel of ischemic heart disease. Antioxidants have potent therapeutic effects on both ischemic heart disease and ischemic-reperfusion injury. Information on the effect of PO on ischemic-reperfusion injury is, however, lacking. In the present study, the effect of dietary palm olein oil on oxidative stress associated with IRI was investigated in an isolated rat heart model. Wistar rats (150-200 gm) of either sex were divided into three different groups (n = 16). Rats were fed with palm olein oil supplemented commercial rat diet, in two different doses [5% v / w (PO 5) and 10% v / w (PO 10) of diet] for 30 days. Control rats (C) were fed with normal diet. After 30 days, half the rats from each group were subjected to in vitro myocardial IRI (20 min of global ischemia, followed by 40 min of reperfusion). Hearts from all the groups were then processed for biochemical and histopathological studies. One way ANOVA followed by Bonferroni test was applied to test for significance and values are expressed as mean +/- SE (p < 0.05). RESULTS: There was a significant increase in myocardial catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities with no significant change in myocardial thiobarbituric acid reactive substances (TBARS) only in group PO 5 as compared to group C. There was no light microscopic evidence of tissue injury. A significant rise in myocardial TBARS and depletion of myocardial endogenous antioxidants (SOD, CAT and GPx) along with significant myocyte injury was observed in control rats subjected to ischemia-reperfusion (C IR). Hearts from palm olein oil fed rats subjected to ischemia-reperfusion (PO 5 IR and PO 10 IR) were protected from increase in TBARS and depletion of endogenous antioxidants as compared to C IR group. No significant myocyte injury was present in the treated groups. CONCLUSIONS: The present study demonstrated for the first time that dietary palm olein oil protected rat heart from oxidative stress associated with ischemic-reperfusion injury. PMID- 15535881 TI - Rationale and design of a multicenter randomized controlled trial on a 'minimal intervention' in Dutch army personnel with nonspecific low back pain [ISRCTN19334317]. AB - BACKGROUND: Researchers from the Royal Netherlands Army are studying the potential of isolated lumbar extensor training in low back pain in their working population. Currently, a randomized controlled trial is carried out in five military health centers in The Netherlands and Germany, in which a 10-week program of not more than 2 training sessions (10-15 minutes) per week is studied in soldiers with nonspecific low back pain for more than 4 weeks. The purpose of the study is to investigate the efficacy of this 'minimal intervention program', compared to usual care. Moreover, attempts are made to identify subgroups of different responders to the intervention. METHODS: Besides a baseline measurement, follow-up data are gathered at two short-term intervals (5 and 10 weeks after randomization) and two long-term intervals (6 months and one year after the end of the intervention), respectively. At every test moment, participants fill out a compound questionnaire on a stand-alone PC, and they undergo an isometric back strength measurement on a lower back machine. Primary outcome measures in this study are: self-assessed degree of complaints and degree of handicap in daily activities due to back pain. In addition, our secondary measurements focus on: fear of movement/(re-) injury, mental and social health perception, individual back extension strength, and satisfaction of the patient with the treatment perceived. Finally, we assess a number of potential prognostic factors: demographic and job characteristics, overall health, the degree of physical activity, and the attitudes and beliefs of the physiotherapist towards chronic low back pain. DISCUSSION: Although a substantial number of trials have been conducted that included lumbar extension training in low back pain patients, hardly any study has emphasized a minimal intervention approach comparable to ours. For reasons of time efficiency and patient preferences, this minimal sports medicine approach of low back pain management is interesting for the population under study, and possibly for comparable working populations with physical demanding job activities. PMID- 15535882 TI - Conservative treatment in patients with an acute lumbosacral radicular syndrome: design of a randomised clinical trial [ISRCTN68857256]. AB - BACKGROUND: The objective is to present the design of randomised clinical trial (RCT) on the effectiveness of physical therapy added to general practitioners management compared to general practitioners management only in patients with an acute lumbosacral radicular syndrome (also called sciatica). METHODS/DESIGN: Patients in general practice diagnosed with an acute (less than 6 weeks) lumbosacral radicular syndrome and an age above 18 years are eligible for participation. The general practitioners treatment follows their clinical guideline. The physical therapy treatment will consist of patient education and exercise therapy. The primary outcome measure is patients reported global perceived effect. Secondary outcome measures are severity of complaints, functional status, health status, fear of movement, medical consumption, sickness absence, costs and treatment preference. The follow-up is 52 weeks. DISCUSSION: Treatment by general practitioners and physical therapists in this study will be transparent and not a complete "black box". The results of this trial will contribute to the decision of the general practitioner regarding referral to physical therapy in patients with an acute lumbosacral radicular syndrome. PMID- 15535883 TI - A genomic timescale of prokaryote evolution: insights into the origin of methanogenesis, phototrophy, and the colonization of land. AB - BACKGROUND: The timescale of prokaryote evolution has been difficult to reconstruct because of a limited fossil record and complexities associated with molecular clocks and deep divergences. However, the relatively large number of genome sequences currently available has provided a better opportunity to control for potential biases such as horizontal gene transfer and rate differences among lineages. We assembled a data set of sequences from 32 proteins (approximately 7600 amino acids) common to 72 species and estimated phylogenetic relationships and divergence times with a local clock method. RESULTS: Our phylogenetic results support most of the currently recognized higher-level groupings of prokaryotes. Of particular interest is a well-supported group of three major lineages of eubacteria (Actinobacteria, Deinococcus, and Cyanobacteria) that we call Terrabacteria and associate with an early colonization of land. Divergence time estimates for the major groups of eubacteria are between 2.5-3.2 billion years ago (Ga) while those for archaebacteria are mostly between 3.1-4.1 Ga. The time estimates suggest a Hadean origin of life (prior to 4.1 Ga), an early origin of methanogenesis (3.8-4.1 Ga), an origin of anaerobic methanotrophy after 3.1 Ga, an origin of phototrophy prior to 3.2 Ga, an early colonization of land 2.8-3.1 Ga, and an origin of aerobic methanotrophy 2.5-2.8 Ga. CONCLUSIONS: Our early time estimates for methanogenesis support the consideration of methane, in addition to carbon dioxide, as a greenhouse gas responsible for the early warming of the Earths' surface. Our divergence times for the origin of anaerobic methanotrophy are compatible with highly depleted carbon isotopic values found in rocks dated 2.8-2.6 Ga. An early origin of phototrophy is consistent with the earliest bacterial mats and structures identified as stromatolites, but a 2.6 Ga origin of cyanobacteria suggests that those Archean structures, if biologically produced, were made by anoxygenic photosynthesizers. The resistance to desiccation of Terrabacteria and their elaboration of photoprotective compounds suggests that the common ancestor of this group inhabited land. If true, then oxygenic photosynthesis may owe its origin to terrestrial adaptations. PMID- 15535884 TI - Omega-3 fatty acids and major depression: a primer for the mental health professional. AB - Omega-3 fatty acids play a critical role in the development and function of the central nervous system. Emerging research is establishing an association between omega-3 fatty acids (alpha-linolenic, eicosapentaenoic, docosahexaenoic) and major depressive disorder. Evidence from epidemiological, laboratory and clinical studies suggest that dietary lipids and other associated nutritional factors may influence vulnerability and outcome in depressive disorders. Research in this area is growing at a rapid pace. The goal of this report is to integrate various branches of research in order to update mental health professionals. PMID- 15535885 TI - Methodology for evaluating Insite: Canada's first medically supervised safer injection facility for injection drug users. AB - Many Canadian cities are experiencing ongoing infectious disease and overdose epidemics among injection drug users (IDUs). In particular, Human Immunodeficiency Virus (HIV) and hepatitis C Virus (HCV) have become endemic in many settings and bacterial and viral infections, such as endocarditis and cellulitis, have become extremely common among this population. In an effort to reduce these public health concerns and the public order problems associated with public injection drug use, in September 2003, Vancouver, Canada opened a pilot medically supervised safer injecting facility (SIF), where IDUs can inject pre obtained illicit drugs under the supervision of medical staff. The SIF was granted a legal exemption to operate on the condition that its impacts be rigorously evaluated. In order to ensure that the evaluation is appropriately open to scrutiny among the public health community, the present article was prepared to outline the methodology for evaluating the SIF and report on some preliminary observations. The evaluation is primarily structured around a prospective cohort of SIF users, that will examine risk behavior, blood-borne infection transmission, overdose, and health service use. These analyses will be augmented with process data from within the SIF, as well as survey's of local residents and qualitative interviews with users, staff, and key stakeholders, and standardised evaluations of public order changes. Preliminary observations suggest that the site has been successful in attracting IDUs into its programs and in turn helped to reduce public drug use. However, each of the indicators described above is the subject of a rigorous scientific evaluation that is attempting to quantify the overall impacts of the site and identify both benefits and potentially harmful consequences and it will take several years before the SIF's impacts can be appropriately examined. PMID- 15535887 TI - Psychometric performance of an assessment scale for strain in nursing care: the M NCAS. AB - BACKGROUND: Multiple instruments exist to measure dementia behaviors, but the nursing staff perspective on those behaviors and their level of burden has not been well measured. The goal of this study was to examine the psychometric performance of the Modified Nursing Care Assessment Scale (M-NCAS), a 28-item nurse rating of burden associated with care for institutionalized individuals with dementia. Nurses rate items in terms of extent to which the behavior or characteristic is present ("attitude" domain), and extent to which it is a burden ("strain" domain). METHODS: Data from 282 patients enrolled in a 12-week, double blind, randomized clinical trial comparing risperidone treatment to placebo was used to evaluate M-NCAS item performance, internal consistency reliability, and construct validity. Empirical subscales were identified via exploratory factor analysis (EFA). RESULTS: Four poorly-performing items were deleted from further analyses. EFA identified 3 "attitude" subscales and 5 "strain" subscales. Cronbach's alphas were 0.65 and above. Correlation with the Cohen-Mansfield Agitation Inventory and the BEHAVE-AD, clinical ratings of dementia behaviors, were low to moderate. CONCLUSION: The M-NCAS provides a valid and reliable means of obtaining care burden ratings from formal caregivers in long-term care, and provides a method for evaluating dementia interventions from the perspective of nursing staff. PMID- 15535886 TI - Role of tyrosine phosphorylation in sperm capacitation / acrosome reaction. AB - Capacitation is an important physiological pre-requisite before the sperm cell can acrosome react and fertilize the oocyte. Recent reports from several laboratories have amply documented that the protein phosphorylation especially at tyrosine residues is one of the most important events that occur during capacitation. In this article, we have reviewed the data from our and other laboratories, and have constructed a heuristic model for the mechanisms and molecules involved in capacitation/acrosome reaction. PMID- 15535888 TI - The development of a new measure of quality of life for young people with diabetes mellitus: the ADDQoL-Teen. AB - BACKGROUND: This study evaluated the psychometric properties of the ADDQoL-Teen, an innovative individualised, patient-centred questionnaire measuring perceived impact of diabetes mellitus on quality of life (QoL) of teenagers. Respondents rate all 30 life domains for frequency, and personally applicable domains for 'bother'. Two overview items measure present QoL and diabetes-dependent QoL. ADDQoL-Teen design was based on the ADDQoL (for adults with diabetes). METHODS: Interviews and discussion groups were conducted with 23 teenagers aged 13-16 years, during work to design the ADDQoL-Teen. The new questionnaire was then completed by 152 young people, (mean age 16.4 +/- 2.4 years), attending diabetes clinics at six UK centres. RESULTS: Five domains detracted from the measure's reliability and factor structure, four of which were analysed separately and one deleted. The 25-domain ADDQoL-Teen had high internal consistency reliability [Cronbach's alpha = 0.91, (N = 133)] and could be summed into an overall Average Weighted Impact score. There were two subscales: a 10-item Impact-Self subscale (measuring impact of diabetes and its treatment on the individual) and a 15-item Impact-Other subscale (measuring impact on interactions with others and the external world). Both subscales had good internal consistency reliability, [Cronbach's alpha coefficients of 0.82 (N = 142) and 0.88 (N = 138) respectively]. Domains reported as most severely (and negatively) impacted by diabetes were (mean weighted impact +/- SD): lie in bed (-3.68 +/- 3.41), interrupting activities (-3.5 +/- 3.23), worry about the future (-3.45 +/- 3.28), career (-3.43 +/- 3.15) and sweets (-3.24 +/- 3.24), (maximum range -9 to +3). Analysis of the overview items showed that although 72.5% considered that their present QoL was good or brilliant, 61.8% felt that having diabetes had a negative impact on QoL, but 35.6% reported no impact and 2.6% reported a positive impact on QoL. CONCLUSIONS: The ADDQoL-Teen is a new measure of perceived impact of diabetes and its treatment on QoL of teenagers. It will help healthcare professionals and parents consider QoL issues as well as medical outcomes when caring for young people with diabetes. It may be used in clinical trials and for routine clinical monitoring in a context of continuing evaluation. PMID- 15535889 TI - The role of d-dimer as first marker of thrombophilia in women affected by sterility: implications in pathophysiology and diagnosis of thrombophilia induced sterility. AB - BACKGROUND: D-dimer is considered a marker of hypercoagulable state and of endogenous fibrinolysis, so increased d-dimer is detectable in patients affected by thrombosis. Yet, several studies showed that also infertility, in particular secondary infertility due to recurrent fetal losses, has been often related to thrombotic events, in particular in women carrying thrombotic risk factors such as inherited thrombophilia (MTHFRC677T, PTHRA20210G, Factor V Leiden polimorphisms and/or inhAfter this screening we selected 39erited protein C, protein S, AT III deficiency) or acquired thrombophilia (primary antiphospholipid syndrome, acquired protein C, protein S, AT III deficiency, drugs induced thrombophilia). However, because its high predictive negative value in case of suspected thrombosis, increased d-dimer has been often associated to subclinical thrombophilia. The aim of this study is to investigate the role of d-dimer as first marker of thrombophilia in women affected by unexplained infertility and subsequently to search the cause of increased d-dimer, such as inherited and/or acquired thrombophilia. PATIENTS AND METHODS: We selected 79 patients with unexplained primary or secondary infertility. We excluded 40 patients affected by hydrosalpinx, uterine fibroids, uterine malformations, endocrinological and immunological diseases, luteal insufficiency, cytogenetical alterations. All remaining 39 patients were tested for d-dimer and divided in two groups: the patients of group A (25 patients) showed increased plasma d-dimer, in group B were included 14 patients with normal plasma level of d-dimer. After this step all 39 patients were screened for MTHFRC677T, PTHRA20210G, Factor V Leiden polimorphisms, protein C, protein S, AT III, anticardiolipin IgM and IgG, lupus anticoagulant. In the control group were included 15 age matched women without sterility problems referred to our outpatient's section of vascular medicine for suspected deep venous thrombosis.Statistical analysis was based on chi2 test, differences were considered to be significant if p < 0.05. RESULTS: D-dimer was increased in 25/39 and 20/25 showed inherited/acquired thrombophilia while patients with normal d-dimer showed inherited/acquired thrombophilia in 7/14 (p: < 0.05, s). DISCUSSION: D-dimer is a well known marker of hypercoagulable state, in particular its high predictive negative value in case of suspected thrombosis has been recognised by several reports. Yet, increased d-dimer has been identified also for subclinical thrombophilia besides for vascular thrombosis. Our data, in fact, for the first time suggest an interesting role of d-dimer to identify women affected by unexplained primary or secondary infertility and thrombophilia. So, probably there is a role for d-dimer in these subjects for its predictive positive value. Of course, further data on large based population are needed to confirm our results, because these findings may speed up a diagnostic screening in these patients also for a good cost/effectiveness of this test. PMID- 15535890 TI - Rate of first recorded diagnosis of autism and other pervasive developmental disorders in United Kingdom general practice, 1988 to 2001. AB - BACKGROUND: There has been concern that the incidence of autism and other pervasive developmental disorders (PDDs) is increasing. Previous studies have been smaller, restricted to autism (excluding other pervasive developmental disorders such as Asperger's syndrome), included boys only, or have not been based on a national sample. We investigated time trends in the rates of diagnosis of pervasive developmental disorders. METHODS: We analysed the rates of first diagnosis of pervasive developmental disorders among people registered with a practice contributing to the United Kingdom General Practice Research Database during the period 1988 to 2001. We included 1410 cases from over 14 million person-years of observation. The main outcome measures were rates of diagnosis of pervasive developmental disorders by year of diagnosis, year of birth, gender and geographical region. RESULTS: The rate increased progressively from 0.40/10,000 person-years (95% CI 0.30 to 0.54) in 1991 to 2.98/10,000 (95% CI 2.56 to 3.47) in 2001. A similar change occurred in the age standardised incidence ratios, from 35 (95% CI: 26-47) in 1991 to 365 (95% CI: 314-425) in 2001. The temporal increase was not limited to children born during specific years nor to children diagnosed in a specific time period. The rate of diagnosis of PDDs other than autism rose from zero for the period 1988-1992 to 1.06/10,000 person-years in 2001. The rate of diagnosis of autism also increased but to a lesser extent. There was marked geographical variation in rates, with standardised incidence ratios varying from 66 for Wales to 141 for the South East of England. CONCLUSIONS: Better ascertainment of diagnosis is likely to have contributed to the observed temporal increase in rates of diagnosis of PDD, but we cannot exclude a real increase. PMID- 15535892 TI - [Mechanical ventilation strategies in patients with acute cardiogenic pulmonary edema]. PMID- 15535891 TI - Review on "Atkins Diabetes Revolution: The Groundbreaking Approach to Preventing and Controlling Type 2 Diabetes" by Mary C. Vernon and Jacqueline A. Eberstein. PMID- 15535893 TI - [Application of recombinant human activated protein C with blood product administration and deep vein thrombosis in severe sepsis]. PMID- 15535894 TI - [Clinical analysis of multiple organ dysfunction syndrome in patients suffering from SARS]. AB - OBJECTIVE: To analyze the features of organ dysfunction in severe acute respiratory syndrome (SARS) and the factors inducing multiple organ dysfunction syndrome (MODS) in these patients. METHODS: The clinical data of 135 SARS patients and 13 patients with severe SARS complicated by MODS were studied retrospectively. RESULTS: SARS could result in organ dysfunction. Its incidence rate was high, the patients' condition was severe, and the mortality was higher. SARS could end in MODS and the mortality was 92.3x. The main dysfunctioning organs were the lung (76.9x), the immune system (92.3x), the cardiovascular system (30.8x), the blood system (61.5x), the liver (38.5x), and the kidney (53.8x). CONCLUSION: Dysfunction of organs are closely correlated with prognosis of SARS. The causes of MODS in SARS are probably injury to the immune system and systemic inflammatory response syndrome. PMID- 15535896 TI - [Effect of intestinal ischemia/reperfusion injury on protein and mRNA levels of Leptin in rats]. AB - OBJECTIVE: To investigate the effect of intestinal ischemia/reperfusion injury on Leptin levels in serum and adipose tissue, and evaluate the potential role of Leptin in acute inflammatory response. METHODS: An intestinal ischemia/reperfusion injury model of rats was reproduced. Serum and adipose tissue Leptin concentrations and Leptin mRNA expression in adipose tissue were determined by a highly-sensitive murine Leptin radioimmunoassay and reverse transcription-polymerase chain reaction (RT-PCR), respectively. RESULTS: Compared with animals prior to injury, serum Leptin level decreased significantly in ischemia for 60 minutes followed by reperfusion for 30 minutes (I60R30), but it tended to increase in I60R150 and significantly increased in I60R360. Compared with sham group, serum Leptin level tended to increase in I60R240 and elevated significantly in I60R360; Leptin levels in adipose tissue significantly decreased in I60R30 and I60R90, while they increased significantly in I60R360. Compared with sham group, Leptin mRNA levels elevated significantly in I60R30, I60R240 and I60R360, but lowered significantly in I60R150. CONCLUSION: Leptin has a time dependent response to acute inflammatory stimuli such as intestinal ischemia/reperfusion injury, and it migh play a role as an inflammatory cytokine. PMID- 15535897 TI - [Protective effect and mechanism of morphine on acute myocardial ischemia/reperfusion injury in rats]. AB - OBJECTIVE: To explore the protective effect of morphine and its mechanism on acute myocardial ischemia/reperfusion (AMIR) injury in rats, by the method of detecting calcitonin gene-related peptide (CGRP) and endothelin-1 (ET-1) levels, as well as myocardial infarct size. METHODS: Forty SD rats were randomly divided into four groups: ischemia/reperfusion group (n=10), morphine preconditioning group (n=10), morphine and naloxone hydrochloride group (n=10), and normal controls (n=10). The animal model of AMIR was established in rats. The left anterior descending branch (LAD) of rat coronary was tied and un-tied. Animals were then sacrificed and hearts were harvested to determine myocardial infarct size by 2, 3, 5-triphenyl tetrazolium chloride (TTC). Radioimmunoassay was used to detect CGRP and ET-1 levels in plasma, and routine method was used to measure creatine kinase isoenzyme (CK-MB) in serum. RESULTS: Plasma ET-1 and CGRP levels were increased significantly than that in normal controls in acute myocardial infarction (AMI) at 10 minutes of LAD tied (all P<0.01). Plasma ET-1 and CK-MB levels in morphine preconditioning group in AMIR at 45 hours of reperfusion were decreased significantly as compared with that in the same group in AMI at 10 minutes, and myocardial infarct size decreased significantly (all P<0.01), while, plasma CGRP levels were markedly increased. Significant differences in those parameters were found between morphine preconditioning group and morphine combined with naloxone hydrochloride group (all P<0.01). CONCLUSION: Intravenous morphine has protective effects on AMI by increased plasma CGRP level, decreased plasma ET-1 level, and reduced myocardial infarct size. PMID- 15535899 TI - [Effect of SARS-associated coronavirus on peripheral blood picture and liver function]. AB - OBJECTIVE: To study peripheral blood picture and liver function of severe acute respiratory syndrome (SARS), and to accumulate more experience in prevention and treatment this syndrome. METHODS: The peripheral blood picture and liver function of 169 cases with SARS were analyzed. These patients were admitted to five hospitals in Taiyuan from March to May, 2003. RESULTS: Above 90.00% patients had normal or low white blood cell (WBC) count on the first day. From then on, during the second to third week, about 40.00% of the patients showed high WBC count average: (13.42+/-2.87)x10(9)/L at 15 th day and (13.68+/-3.26)x10(9)/L at 21 st day) and neutrophil (0.809+/-0.063 at 15 th day and 0.805+/-0.061 at 21 st day), and 21.97%-52.15% cases with low lymphocyte count (0.111+/-0.044-0.134+/-0.040). About 4.44%-12.76% of patients developed low platelet and 4.20%-33.33% of cases with low hemoglobin level. And 32.76%-62.50% of cases were found to have high alanine aminotransferase (above 40 U/L), 13.04%-40.00% cases with high aspartate aminotransferase (above 40 U/L) and 40.35%-72.00% cases with low albumin content. But serum total protein remained normal during the whole clinical course. CONCLUSION: SARS cases may develop leucocytosis, low lymphocyte count, low platelet count, low hemoglobin level and abnormal liver function. PMID- 15535901 TI - [Comparison of serum biochemical features between SARS and other viral pneumonias]. AB - OBJECTIVE: To identify blood chemistry changes in coronavirus of severe acute respiratory syndrome (SARS). METHODS: Biochemical changes in SARS patients were summarized and compared with other viral pneumonias. Serum total protein (TP), albumin (Alb), total cholesterol (TC), triglyceride (TG), calcium (Ca), ferrum (Fe), lactate dehydrogenase (LDH), creatine kinase (CK), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) between SARS and other viral pneumonias were examined by Roche Diagnostics assay, HITACHI7600 automatic analyzer. Clinically confirmed SARS patients, patients with other viral pneumonias, and healthy controls were included in the study. RESULTS: Compared with healthy person, the levels of serum TC, Fe, Ca, Alb were significantly lowered (P<0.05 or P<0.01), while the activity of LDH, CK, ALT, AST were elevated, the increase of CK and the decrease of Fe were the most significant (P<0.05 or P<0.01), the changes of TP and TG were not obvious. In the other viral pneumonias patients, ALT, AST, LDH were elevated slightly than those of healthy person, while Fe, Ca, Alb, TC, CK were a little reduced, but there was no significant difference between the two groups. In convalescent stage, all the tests were returned to normal ranges except ALT, AST were still elevated in SARS patients. CONCLUSION: The changes in serum biochemistry are more marked in SARS patients compared with patients suffering from other viral pneumonias, the decrease of Fe as well as the inhibition of TC may be caused by the treatment of anti-virus. PMID- 15535903 TI - [A preliminary report of SARS coronavirus specific RNA in SARS convalescents and medical person]. AB - OBJECTIVE: To observe the method of fluorescent-polymerase chain reaction (F-PCR) and gene-hip technique in detecting coronavirus in severe acute respiratory syndrome (SARS) and its value for clinical application. METHODS: Serum of 60 SARS patients, 20 samples of serum and 20 samples of gargling fluid of medical staff of the "Fever Clinic", and one cDNA specimen obtained from one SARS suspect patient were examined with F-PCR diagnosis kit and gene-chip technique for SARS coronavirus. RESULTS: Three methods, including DR Chip and two F-PCR kit from Zhongshan College Da'an gene stock company and Shanghai Fortune industrial joint stock Co, Ltd, were used, and the results were all negative for all specimens, except one cDNA specimen, which was obtained from one SARS suspect patient, virus could be amplified by F-PCR. CONCLUSION: Special RNA fragment of SARS virus has not been detected in gargling fluid and serum of SARS convalescents and medical staff. PMID- 15535905 TI - [Clinical study on respiratory mechanics of synchronized intermittent mandatory ventilation in premature infants with respiratory failure]. AB - OBJECTIVE: To evaluate the clinical significance of synchronized intermittent mandatory ventilation (SIMV) in prematures infants necessitating assistant ventilation. METHODS: Forty-two premature infants were studied in whom SIMV was used in 24 and intermittent mandatory ventilation (IMV) was used in 18. The parameters of fractional concentration of inspired oxygen (FiO(2)), peak inspiratory pressure (PIP), expiratory tidal volume (V(Texp)), oxygen index (OI), compliance of the respiratory system (Crs) and airway resistance (Raw) were compared. The frequency of administration of sedatives and incidence rate of complications were also compared between two groups. RESULTS: In SIMV group, PIP, V(Texp), Raw and Crs were significant better after 2 hours ventilation (all P<0.05). There were significant difference in FiO(2) after 12 hours and OI at 24 hours in two groups. They were better in SIMV group. Sedatives were less used in SIMV group than IMV group (P<0.05). CONCLUSION: In premature infants, SIMV could reduce FiO(2), PIP, OI, Raw and improve V(Texp), Crs more rapidly when assistant ventilation is necessary. The sedatives are used less in patients on SIMV. PMID- 15535906 TI - [Analysis of prognosis assessment with acute physiology and chronic health evaluation III, multiple organ dysfunction score and sequential organ failure assessment for the postoperative patients in cardiovascular surgery]. AB - OBJECTIVE: To analyze the clinical significance of the prognosis assessment with acute physiology and chronic health evaluation III (APACHEIII), multiple organ dysfunction score (MODS) and sequential organ failure assessment (SOFA) for postoperative patients in cardiovascular surgery. METHODS: Prognosis of 68 patients undergoing cardiovascular operation from February 2004 to June 2004 in our ward was assessed by APACHE III, MODS and SOFA. Then the scores were calculated and compared everyday while they stayed in Cardiac Surgery Intensive Care Unit (CSICU). RESULTS: The scores of three methods showed no significant difference from one another in the first three postoperative days, but were significantly higher than all the scores calculated just after the entrance to the CSICU (A0, M0, S0; all P<0.01). There were no differences in the changes of APACHE III, MODS and SOFA(DeltaA, DeltaM and DeltaS) in the first three postoperative days respectively, although they all showed a decreasing tendency. APACHE III scores were positively correlated with MODS although the correlation were diminishing (P<0.01 at first day but P<0.05 at third day), while they were positively correlated with SOFA only in the first two days (both P<0.01). The MODS was positively correlated with SOFA at various time points (P<0.001). The length of stay in CSICU was positive correlated with A0 and maximum of APACHEIII (Amax, P<0.05), and M0, maximum of MODS (Mmax) as well as S0, maximum of SOFA (Smax, P<0.001), respectively. CONCLUSION: For the patients who have undergone cardiovascular operation, A0 could assess the prognosis fairly precisely, but MODS and SOFA assessment seem to be better than APACHE III. Individual Smax and kinetic DeltaS might be the most suitable indexes for cardiovascular surgery. PMID- 15535907 TI - [Study on pulmonary fibrosis and state of angiogenesis in acute lung injury]. PMID- 15535908 TI - [Effects of infused intravenous ice-cold normal saline on the cerebral perfusion pressure of rabbits with focal cerebral ischemia]. PMID- 15535909 TI - [Influence of macrophage colony-stimulating factor and neopterin on acute coronary syndrome]. PMID- 15535910 TI - [Intra-arterial microorganism infection in patients with acute myocardial infarction and unstable angina pectoris]. PMID- 15535912 TI - [Detection of bacteria producing extended -spectrum beta-lactamases and the analysis of drug resistance]. PMID- 15535914 TI - [Value of intra-abdominal pressure monitoring in the prevention and treatment of multiple organ dysfunction syndrome]. PMID- 15535923 TI - [Two year follow-up of inpatients with alcohol-related diseases]. AB - BACKGROUND AND OBJECTIVE: Our objective was to study the outcome and compliance of both medical and alcoholism treatment in alcoholics discharged from a general hospital. PATIENTS AND METHOD: This was a 2 years prospective follow up study of a cohort of 100 inpatients referred to the Alcohol Unit during the first semester of 1998. RESULTS: After 2 years, 9 patients had died and 14 had been referred to other hospitals. 15% of patients refused medical treatment from the beginning and alcoholism treatment was initially refused by 31% of alcoholics. At 2 years of follow up, compliance rates were 46% for medical treatment and 10% for alcoholism treatment. All patients under treatment for alcoholism were also good compliers of medical treatment. Survival analysis for alcoholism treatment at 2 years was 19%, with higher rates in patients with psychosocial problems and previous alcohol consultations. The probability of being alive was 89%. CONCLUSIONS: Few alcoholics are detected. Most of them display high levels of severity and poor prognosis, with high mortality rates. Compliance with follow up treatment is low. There is a clear correlation between compliance of alcoholism and medical treatment. PMID- 15535924 TI - [Changes in CD4+ and CD8+ T-cell subsets in HIV-infected children differently correlate with viral replication and thymic function]. AB - BACKGROUND AND OBJECTIVE: We intended to study the relationship between T-cell subsets with plasmatic detectable viral load (VL) and T-receptor excision circles (TREC). PATIENTS AND METHOD: Twenty HIV-infected children on highly active antiretroviral therapy (HAART) were recruited in a 1-year longitudinal retrospective study. We analyzed the relationship between changes in peripheral blood T-cell subsets, VL and TREC markers by lineal regression. RESULTS: Memory and activated CD4+ T-cells increases had a negative association with log10 TRECs increases. However, naive CD4+ T-cells increases had a positive association with log10 TRECs increases. In contrast, memory, activated and effector CD8+ T-cells increases positively correlated with log10 VL increases. On the other hand, naive CD8+ T-cells increases had a negative association with log10 VL increases. CONCLUSIONS: CD4+ and CD8+ T-cells subsets change in a different way as a response to the changes produced by HAART in HIV vertically infected children. CD4+ T-cells are more dependent on thymic function and CD8+ T-cells are more dependent on viral replication. Thus, the decline in cellular activation would allow the production of more naive T-cells by the thymus. PMID- 15535925 TI - [Influenza vaccination in healthcare workers. Strategies to achieve compliance in a general hospital]. AB - BACKGROUND AND OBJECTIVE: Since healthcare workers can transmit the influenza virus to high risk patients, they should be vaccinated prior to the influenza season. In this study we describe the influenza vaccination coverage in healthcare workers in a general hospital and study its evolution over the last years. SUBJECTS AND METHOD: Descriptive epidemiological study to determine the anti-influenza vaccination coverage in healthcare workers in a hospital during 2002-2003 and 2003-2004 seasons. The variables studied in each season were sex, age and professional category. RESULTS: During each of the campaigns, 1,215 and 2,287 workers were vaccinated. The vaccination coverage increased significantly (p < 0.01) (20.2% in 2002-2003 and 38% in 2003-2004). Men were vaccinated more than women although the coverage increase was significant in both sexes (p < 0.01). During the 2002-2003 season, workers older than 50 years (24.4%) had the greatest coverage, while during 2003-2004 those older than 31 years (48.7%) were the ones with more coverage. By professional categories, the medical staff (33.8% in 2002-2003 and 59.9% in 2003-2004) had a greater vaccination acceptance. The coverage increase during these two years was significant in all healthcare workers (p < 0.01). CONCLUSIONS: A significant increase in the vaccination coverage has been observed during the two seasons studied. This fact can be related to the changes introduced in the informative strategies and the performance of more active vaccination campaigns which connect vaccination to employment areas. PMID- 15535926 TI - [Atorvastatin lowers C-reactive protein in dislipemic patients with type 2 diabetes mellitus]. AB - BACKGROUND AND OBJECTIVE: Type 2 diabetes mellitus is associated with an augmented risk for cardiovascular disease. The levels of C-reactive protein (CRP), the prototypic marker of inflammation, are associated with an increased risk for cardiovascular events. The statins have direct anti-inflammatory effects. Thus, we tested the effects of atorvastatin on levels of CRP on patients with type 2 diabetes. PATIENTS AND METHOD: We evaluated CRP in baseline and 6 months after onset of 20 mg daily atorvastatin therapy of 30 patients with type 2 diabetes with hyperlipidemia. Clinical and biochemical data were obtained. RESULTS: CRP-levels were significantly decreased after treatment with atorvastatin compared with baseline (median change: -4,99 mg/l; p < 0.001). We observed an correlation between CRP baseline with body mass index (r = 0.429; p = 0.018), serum fibrinogen (r = 0.607; p = 0.001) and microalbuminuria (r = 0.470; p = 0.01). Conversely, there was no significant correlation between CRP baseline with LDL cholesterol. The CRP reduction was significantly correlated with fasting glucose (r = -0.457; p = 0.019) and glycosylated hemoglobin at 6 months (r = 0.421; p = 0.03). CONCLUSIONS: These results confirm findings from previous studies that atorvastatin reduce CRP levels in a largely LDL cholesterol independent manner. PMID- 15535927 TI - [Alcoholism: a subject in standby]. PMID- 15535928 TI - [Therapeutical perspectives in systemic lupus erythematosus]. PMID- 15535929 TI - [Haplotype of the BRCA2 6857delAA mutation in 4 families with breast/ovarian cancer]. AB - BACKGROUND AND OBJECTIVE: It is estimated that 5-10% of all breast cancers are hereditary, mainly are due to germline mutations in the BRCA1 and BRCA2 genes. PATIENTS AND METHOD: A BRCA2 screening was carried out in familial breast/ovarian cancer at two centres in Spain and Chile. The 6857delAA mutation was identified in 3 Spanish families and one Chilean, all of them with Spanish ancestors. The BRCA2 haplotype of the 6857delAA carriers was analyzed using five microsatellite markers flanking the BRCA2 gene, spanning a region of 6 cM: cen-D13S260, D13S1698, (BRCA2), D13S171, D13S310 and D13S267-tel. RESULTS: Two families shared the allelic variants of the 5 microsatellites studied. Markers D13S260 and D13S267 differed in one allele in two families, respectively. The defined haplotype was absent in non-carriers from these families, and was not detected in 100 control chromosomes without the mutation. CONCLUSIONS: Our results suggest the existence of a common ancestry with the mutation originating in the Northeast of Spain. Given the migratory movements from Spain to Latin America, the screening of recurrent Spanish mutations can be useful in establishing a more rational and cost-effective analysis in such populations. PMID- 15535930 TI - [Is it ethical conducting placebo-controlled clinical trials as part of the development of new drugs for the treatment of major depressive disorder? (I). Considerations on the value and scientific validity of the study]. PMID- 15535931 TI - [Value of troponins in acute coronary syndrome in patients with renal failure]. AB - Patients with renal insufficiency can have elevations of serum troponin without suspected clinical coronary ischemia. Although cardiovascular disease is the main cause of death in patients with renal failure, the process of elevation of serum troponin is not well known. Troponin T is more frequently elevated than troponin I in these patients which leads to uncertainty in the clinical interpretation of results. There are studies suggesting that troponin elevations are associated with a higher risk and increased mortality. To explain the process leading to troponin increases in this kind of pathology and to confirm its usefulness in the diagnosis, evolution and prognosis it would be necessary to carry out more clinical studies monitoring troponin and studying the stratification of risk. PMID- 15535932 TI - [Disseminated Geotrichum capitatum infection in neutropenic patient with acute leukaemia]. PMID- 15535933 TI - [A suicide attempt by subcutaneous injection of mercury]. PMID- 15535934 TI - [Susac syndrome. Differential diagnosis]. PMID- 15535935 TI - [Periodic alternating gaze deviation in hepatic coma]. PMID- 15535936 TI - [Relationship between the peripheral arterial occlusive disease and the infection by Chlamydophila pneumoniae]. AB - BACKGROUND AND OBJECTIVE: The relationship between peripheral arterial occlusive disease (PAOD) and Chlamydophila pneumoniae infection was studied by analyzing clinical samples from 95 patients with PAOD (cases) and 100 controls. PATIENTS AND METHOD: The following investigations were conducted: IgG and IgA against lipopolysaccharide (LPS) and against purified C. pneumoniae-specific antigens from elementary bodies (EB) with ELISA; anti-EB IgG, with MIF; C. pneumoniae DNA in arterial biopsy and peripheral blood leukocyte cells (PBLCs) with heminested PCR; LPS with ELISA; and bacteria culture in HEp-2 cells from arterial biopsy. RESULTS: The percentage of positive results in cases and controls groups for anti LPS IgG was: 21% and 14%, respectively, with no differences; nor were there any differences with IgA (22 and 21%, respectively). However, differences were seen in the anti-EB IgG between cases (74% and 72%, for ELISA and MIF, respectively) and controls (31% and 34%). There were no differences in anti-EB IgA. Bacterial DNA was detected in 67% of atheromatous plaques (cases) vs. 12% of pudendal arteries (controls) (p = 0.0001). No C. pneumoniae DNA and LPS was detected in PBLCs and biopsic samples, respectively; and no C. pneumoniae strain could be recovered by cell culture from cases. CONCLUSIONS: On the basis of our results, PAOD is significantly associated with C. pneumoniae infection through the detection of anti-EB IgG from serum and bacterial DNA from arterial biopsy. PMID- 15535937 TI - [Hypertriglyceridemic acute pancreatitis. Is its clinical course different from lithiasic acute pancreatitis?]. AB - BACKGROUND AND OBJECTIVE: Although hypertriglyceridemia is an accepted cause of acute pancreatitis, its clinical course is not well defined. What is more, lipemic serum may interfere with an accurate pancreatic enzyme measurement and may hinder the diagnosis of acute pancreatitis. The objective was to analyze the clinical performance of hypertriglyceridemic acute pancreatitis and the diagnosis value of the serum measurement of pancreatic enzymes. PATIENTS AND METHOD: 31 demographic, clinical, analytical and radiological data of 19 hypertriglyceridemic acute pancreatitis were retrospectively analyzed and their results were compared with data of 19 lithiasic acute pancreatitis without hypertriglyceridemia. Diagnosis of acute pancreatitis was based on clinical, radiological and/or laparotomical findings. Serum levels of triglycerides higher than 1,000 mg/dl were considered the cause of acute pancreatitis when other causes were excluded. Acute pancreatitis was considered lithiasic when gallstones were identified by ultrasonography, serum levels of triglycerides were lower than 200 mg/dl and there was no history of alcohol consumption. RESULTS: Compared with lithiasic acute pancreatitis, hypertriglyceridemic acute pancreatitis showed more personal and family hypertriglyceridemia (9 vs 0), more previous episodes of pancreatitis (13 vs 2), pancreatitis was more severe (13 vs 5) and had more complications (29 vs 5). There was no mortality in either group. The serum levels of amylase and lipase supported the diagnosis in 26% and 58% of patients with hypertriglyceridemic acute pancreatitis compared with 58% and 79% of patients with lithiasic acute pancreatitis, respectively. Hospital stay was longer in hypertriglyceridemic than in lithiasic acute pancreatitis (24 [45] vs 7.6 [3.1] days; p = NS). Demographic and analytical factors which could permit to predict the severity of acute pancreatitis could not be identified. CONCLUSIONS: Hypertriglyceridemic acute pancreatitis is relapsing and its clinical course is more severe than lithiasic acute pancreatitis. The measurement of amylase and lipase levels is less useful in the diagnosis of hypertriglyceridemic than in lithiasic acute pancreatitis. PMID- 15535938 TI - [Relationship between pesticide exposure and low-grade superficial bladder urothelial carcinoma]. AB - BACKGROUND AND OBJECTIVE: Few studies have been published analyzing the association between pesticides use and the increased risk of developing urothelial cancer of the bladder (UCB). The aim of this work was to investigate, in a geographical area with a high prevalence of UCB (Axarquia, Malaga province, southern Spain) if a) subjects with occupational exposure to pesticides have greater risk of developing UCB and b) there are histopathological differences with regard to UCB in patients without any exposure. PATIENTS AND METHOD: Case control study. During two years (1994-1996) 96 cases of UCB were included. The histologic grade and the depth of the invasion as well as the associated inflammatory infiltrate were analyzed. A questionnaire was answered by the patients. RESULTS: 58.3% of the case group were working with or had history of exposure to pesticides, compared to 40.6% in the control group (OR = 2.04; 95% CI, 1.1-3.6). The risk increased with a greater time of exposure. In exposed subjects, low grade tumors (OR = 2.6; 95% CI, 1.3-5.2) as well as superficial tumors (OR = 2.3; 95% CI, 1.2-4.4) were more frequent. They were more frequently accompanied by a chronic inflammatory infiltrate (OR = 4.5; 95% CI, 1.8-11.1). CONCLUSIONS: The subjects with occupational exposure to pesticide have greater risk of developing UCB, which is directly proportional to the exposure time. In comparison with the population in the zone without exposure, patients who use pesticides present low grade UCB and less invasive tumors more frequently. These facts were not modified when they were adjusted for tobacco consumption. Exposed patients have UCB with chronic (moderate) inflammatory reaction more frequently than the tumors in non-exposed subjects. PMID- 15535939 TI - [Risk factors of mortality in invasive pneumococcal disease]. AB - BACKGROUND AND OBJECTIVE: To analyze the risk factors associated with mortality in invasive pneumococcal disease in the university hospital Fundacion Jimenez Diaz (Madrid, Spain) during 11 years. PATIENTS AND METHOD: We performed a retrospective study of patients attending the emergency service of the hospital from January 1993 to August 2003. We registered data on mortality, clinical and microbiological evolution and relapses. RESULTS: We studied 263 patients with pneumococcal baceteremia and invasive disease caused by Streptococcus pneumoniae (pneumonia, meningitis, sepsis, bacteremia of unknown origin and oligoarthritis). Mortality was 12.5%. Variables associated with mortality in a logistic regression analysis included absence of leukocytosis (p = 0.04), acidosis (p < 0.01), respiratory signs and symptoms (tachypnea, pleuritic pain) (p = 0.02), and neurologic manifestations (decreased consciousness level; (p < 0.01). CONCLUSION: Patients at highest risk of death because of invasive pneumococcal disease are critically ill, with no leukocytosis, with severe respiratory or neurological symptoms and undergoing invasive procedures such as mechanical ventilation and tracheostomy. PMID- 15535940 TI - [Mortality in pneumococcal disease]. PMID- 15535941 TI - [Diagnosis of bronchial eosinophilic inflammation: an important step]. PMID- 15535942 TI - [Legionnaires' disease and HIV infection: an opportunistic infection?]. AB - BACKGROUND AND OBJECTIVE: Legionella infections are not frequent in HIV-infected patients, although clinical manifestations and outcome are particularly severe in this subset. This manuscript analyzes the clinical features and immunological situation of HIV-infected patients with Legionnaires' disease (LD). PATIENTS AND METHOD: The clinical files of HIV-infected patients diagnosed with LD from 1983 to December 2003 were reviewed. The incidence of hospital-acquired Legionella pneumonia (HALP) from 1997-2000 in HIV-infected patients was compared with that of non infected patients. RESULTS: Eighteen patients were included. 72.2% were diagnosed by the Legionella urinary antigen assay. The incidence of HALP in HIV infected and non infected patients was 0.3 and 0.25/1000 admissions/year, respectively (p = 0.42). 83.3% received appropriate antibiotic treatment at the Emergency department. The mean lymphocyte CD4 count was 348.1/microl, 53.8% had an undetectable viral load and 64.7% were on antiretroviral therapy. 72.2% were smokers, 38.8% had cancer and 16.7% were on chemotherapy. 93.8% had cough, 75% dyspnea, 62.5% extrarespiratory symptoms, 76.5% increased AST, 50% increased CK and 56.3% hyponatremia. Moreover, 50% developed bilateral pulmonary infiltrates, 83.3% respiratory failure and 22.2% died. CONCLUSIONS: Although LD is not more frequent in HIV-infected than in non infected patients, its clinical severity suggests that it is an opportunistic infection. PMID- 15535943 TI - [Is it ethical conducting placebo-controlled clinical trials as part of the development of new drugs for the treatment of major depressive disorder? (and II). Benefit/risk ratio and informed consent. Conclusions]. PMID- 15535944 TI - [Histiocytic sarcoma]. PMID- 15535945 TI - [Advance directives and medical practice]. PMID- 15535946 TI - [Hemolytic-uremic syndrome secondary to acute pancreatitis with spontaneous spleen rupture]. PMID- 15535947 TI - [Pulmonary sequestration with systemic coronary arterial supply]. PMID- 15535948 TI - [Cutaneous plasmacytomas following treatment of IgA kappa multiple myeloma]. PMID- 15535949 TI - [Systemic lupus erythematosus syndrome induced by interferon therapy]. PMID- 15535950 TI - [Angina with ST segment elevation during resting phase of dobutamine stress]. PMID- 15535951 TI - [Pulmonary actinomycosis secondary bacteremia in a HIV-infected patient]. PMID- 15535952 TI - Javelin diagrams: applications in veterinary medical decision analysis. AB - This paper introduces javelin diagrams as an innovative way for depicting the results of medical decision analyses. The methods were used to determine whether, and at which values, blood lactate in Belgian White and Blue or maximum tidal volumes in Holstein calves should be measured before deciding to treat or not a calf suffering from the bovine respiratory disease complex. The different alternatives depended upon the probabilities of survival with and without treatment and upon the costs associated with a possible death, the test and the treatment. The chosen alternative was the one with the lowest expected costs. From data collected on the treated calves, the expected costs of measuring lactate (198.01 euro) and tidal volumes (27.38 euro) before deciding to treat or not were lower than the expected costs of directly treating sick Belgian Blue (215.39 euro) and Holstein (51.55 euro) calves, respectively. The treatment should be applied to sick Belgian Blue calves with blood lactate < or = 7.8 mmol/L and to Holstein calves with a maximum tidal volume > or = 1.81 L. At such test values, the treatment expected costs were lower than the expected costs associated with no treatment of calves with other test values. Probabilistic sensitivity analyses showing benefits in treating animals with a positive test (over not treating the animals with a negative test) were mostly invariant to changes in any cost value but were sensitive to uncertainties in probabilities of survival with or without treatment. The javelin diagrams provided a clear visual indication of such results. They depicted how and by how much the benefits were affected by uncertainties in probabilities, they proposed different information values associated with the chosen alternative, and identified directions for further research. PMID- 15535953 TI - In vitro effect of classical swine fever virus on a porcine aortic endothelial cell line. AB - The effect of classical swine fever (CSF) virus on some phenotypic and functional features of an established porcine aortic endothelial cell (AOC) line was investigated. AOC cells show most of the characteristics of primary endothelial cells, avoiding the alterations and senescence that these cells undergo after a few passages in culture. AOC cells were susceptible to CSF virus infection to a high degree, reaching 90% of CSF virus positive cells after 24 h of infection; however as with other porcine susceptible cells, no cytopathic effect could be observed. In these conditions none of the surface molecules studied, including SLA-II MHC antigens, adhesion or co-stimulatory molecules, were altered by virus infection after 24 or 48 h. Functionally CSF virus infection induced a decrease in the pro-coagulant activity of the AOC cells, determined by the increase in the clot formation time shown by the lysates of these cells. This contrasts with the increase observed in the expression of mRNA corresponding to IL-1 alpha and IL-6, two proinflammatory and pro-coagulant cytokines, in CSF virus-infected AOC cells. PMID- 15535954 TI - Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants. AB - DNA amplification techniques offer considerable promise for the identification of Mycoplasma mycoides cluster members. They avoid antigenic cross-reactivity and variability that hamper serological methods. Many sets of primers, specific of these different members and of Mycoplasma putrefaciens, have been proposed. To assess the reliability of some of these PCR tests in routine laboratory diagnostic use, 230 field strains supposed to belong to this group were simultaneously identified by PCR and an antigenic method. The results were well correlated to antigenic identification for M. putrefaciens, but PCR failed to identify respectively 74% and 52% of M. mycoides subsp. mycoides Large Colony type and M. capricolum subsp. capricolum strains. Any identification of M. mycoides subsp. mycoides Small Colony type must be confirmed by two different tests. Difficulties in defining the M. species bovine serogroup 7 were also encountered with both the PCR and immunological methods. The occurrence of putative variable antigen(s) on the mycoplasma surface may explain part of the identification difficulties encountered with the immunological methods. PMID- 15535955 TI - Serum amyloid A and TNF alpha in serum and milk during experimental endotoxin mastitis. AB - A cross-over study was conducted to investigate the effect of intramammarily infused lipopolysaccharide (LPS) on the acute phase reaction in early (EL) and in late (LL) lactation. Nine cows received intramammary injections of 100 microg of Escherichia coli 0111:B4 LPS during EL and LL. The severity of each cows systemic and local signs and change in milk appearance were recorded and scored throughout the experiment. Systemic and local signs were found to be more serious in EL cows. Tumor necrosis factor alpha (TNF alpha) was detected in milk but not in serum. Serum amyloid A (SAA) concentrations increased both in serum and in milk. The milk TNF alpha concentrations peaked at 8 h post-challenge (PC). SAA concentrations started to increase at 8 h PC, and peak concentrations were seen at 32 and 48 h PC in milk and serum, respectively. The milk TNF alpha and SAA seemed to be correlated, being on average higher in EL. Serum SAA concentration was not correlated with milk TNF alpha or SAA, nor with the severity of local or systemic signs, but was correlated with changes in milk appearance. PMID- 15535956 TI - The comparative pathogenicity of strains of eight serovars and untypable strains of Mannheimia haemolytica in experimental pneumonia of sheep. AB - The experimental induction of pneumonic pasteurellosis in groups of conventionally reared lambs by 8 serovars (A1, A2, A6, A7, A8, A9, T10, and A11) and untypable (UT) strains of Mannheimia haemolytica (Mh) were examined and compared. The groups of lambs were inoculated intratracheally with 1.4 x 10(8) +/ 0.6 x 10(8) (mean +/- SD) colony-forming units of the Mh serovars or UT isolates in the 6-hour log phase of growth. The variables measured as indicators of disease severity were clinical score, percentage lung consolidation and microbiological re-isolation. The clinical parameters for each group were computed daily for 6 days post infection and the lambs which died were necropsied while the remaining lambs were killed on day 7 pi and the extent of lung consolidation was measured. Clinically, the mean scores for the M. haemolytica serovars were A1 (6.1), A2 (18.8), A6 (0.5), A7 (17.4) and A9 (8.5). The mean percent lung lesion scores for M. haemolytica serovars were A1 (12.5), A2 (66.3), A6 (5.0), A7 (51.3), A9 (33.8) and A11 (2.5). The percent mean pneumonic lung lesions recorded for groups inoculated with A2, A7 and A9 were significantly (P < 0.05) higher than the extent of lung lesions in the other groups. A statistically significant correlation was observed between clinical scores and the severity of the lung lesions (r = 0.96, P < 0.01). High titres of M. haemolytica were recovered from lung lesions, with 10 to 100 times the number of organisms inoculated being present in the lung lesions of lambs inoculated with serovars A2 and A7. These data indicate that although M. haemolytica serovars A1, A2, A6, A7, A9 and A11 are important primary lung pathogens of lambs, serovars A2, A7, and A9 are to be regarded as highly virulent strains that have a greater predilection than the other serovars for causing pneumonia in lambs. PMID- 15535957 TI - An automated spectrophotometric method for measuring canine ceruloplasmin in serum. AB - An automated method for the determination of ceruloplasmin activity was developed and validated in canine serum. The method is based on the in vitro oxidase activity that this protein shows with substances such as p-phenylenediamine. In order to determine optimum assay conditions, the effects of the substrate concentration, buffer pH, reaction time and EDTA on the reaction were evaluated. The precision of the assay was good with within-run and between-run coefficients of variation lower than 10%. The method measured the ceruloplasmin values in a proportional and linear manner (r = 0.99) with a limit of detection of 0.0007 +/- 0.0001 Delta Abs/min. A temperature of -20 degrees C kept the reagent stable for 30 days. The method is cheap and easy to adapt to any automated biochemical analyser, considerably decreasing the processing time required with the manual method. Additionally it allows to differentiate dogs with pyometra and trauma from clinically healthy dogs. PMID- 15535958 TI - Innate immune response to intramammary infection with Serratia marcescens and Streptococcus uberis. AB - Streptococcus uberis and Serratia marcescens are Gram-positive and Gram-negative bacteria, respectively, that induce clinical mastitis. Once initial host barrier systems have been breached by these pathogens, the innate immune system provides the next level of defense against these infectious agents. The innate immune response is characterized by the induction of pro-inflammatory cytokines, as well as increases in other accessory proteins that facilitate host recognition and elimination of the pathogens. The objective of the current study was to characterize the innate immune response during clinical mastitis elicited by these two important, yet less well-studied, Gram-positive and Gram-negative organisms. The pro-inflammatory cytokine response and changes in the levels of the innate immune accessory recognition proteins, soluble CD14 (sCD14) and lipopolysaccharide (LPS)-binding protein (LBP), were studied. Decreased milk output, induction of a febrile response, and increased acute phase synthesis of LBP were all characteristic of the systemic response to intramammary infection with either organism. Infection with either bacteria similarly resulted in increased milk levels of IL-1 beta, IL-8, IL-10, IL-12, IFN-gamma, TNF-alpha, sCD14, LBP, and the complement component, C5a. However, the duration of and/or maximal changes in the increased levels of these inflammatory markers were significantly different for several of the inflammatory parameters assayed. In particular, S. uberis infection was characterized by the sustained elevation of higher milk levels of IL-1 beta, IL-10, IL-12, IFN-gamma, and C5a, relative to S. marcescens infection. Together, these data demonstrate the variability of the innate immune response to two distinct mastitis pathogens. PMID- 15535959 TI - Biotin-avidin amplified ELISA for detection of antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.). AB - Scabies is a major threat to the well being of mountain-dwelling Bovid hosts, Rupicapra rupicapra and Rupicapra pyrenaica. Severe outbreaks are in progress over a significant part of their distribution area and resource managers demand improved methods to monitor, analyse and possibly forecast the spread and effects of scabies at the population level. An amplified capture enzyme-linked immunosorbent assay was developed to detect antibodies to Sarcoptes scabiei in chamois (Rupicapra spp.) serum. The method used the biotin-avidin amplification system and was validated on a panel of 144 serum samples, of which 40 were obtained from scabietic and 104 from healthy unexposed individuals originating from a scabies-free area. The antigen, a whole body extract of the various developmental stages of S. scabiei, was prepared from mites actively leaving the skin lesions of naturally infested red foxes (Vulpes vulpes). The resulting LAB ELISA was characterised by 93% sensitivity, 97% specificity and a high degree of repeatibility. A single seroreactor was found amongst 32 chamois affected with skin pathologies other than scabies, including infestations by other Acarina (Trombicula spp. and Ixodid ticks). Antibodies to S. scabiei were present in 26 out of 169 sera (15.4%) obtained by clinically healthy chamois within a scabies outbreak area, indicating that asymptomatic infestations by S. scabiei can be revealed by serological methods in the studied Caprinae hosts. PMID- 15535960 TI - Determination of an efficient and reliable method for DNA extraction from ticks. AB - Molecular detection of pathogenic microorganisms in ticks is based on DNA amplification of the target pathogen; therefore, extraction of DNA from the tick is a major step. In this study, we compared three different tick DNA extraction protocols based on an enzymatic digestion by proteinase K followed by DNA extraction by a commercial kit (method 1), or on mortar crushing, proteinase K digestion and phenol/chloroform DNA extraction (method 2) and fine crushing with a beads beater, proteinase K digestion and DNA extraction using a commercial kit (method 3). The absence of PCR inhibitors and the DNA quality were evaluated by PCR amplification of the tick mitochondrial 16S rRNA gene using tick-specific primers. With method 1, 23/30 (77%) of the samples were extracted; with method 2, 30/31 (97%) of the samples were extracted and with method 3, 30/30 (100%) of the samples were extracted. DNA extraction efficiency using method 3 is significantly higher than DNA extraction efficiency using method 1 (100% versus 77%, P < 0.05). There was no significant difference between methods 2 and 3. Method 3 was however more adapted to cohort studies than method 2. This technique was validated for cohort tick DNA extraction and applicable to the treatment of small samples such as nymphs and soft ticks with 100% efficiency. PMID- 15535961 TI - Bovine herpesvirus 1 glycoprotein D expression in bovine upper respiratory tract mediated by a human adenovirus type 5. AB - Bovine herpesvirus 1 glycoprotein D (gD) gene expression by recombinant replication defective human adenovirus type 5 (HAdV-5) was investigated in calves using indirect immunofluorescence microscopy (IIFM), confocal laser scanning microscopy (CLSM) and RT-PCR. One fold intranasal instillation of HAdV-5 expressing gD in the cattle upper respiratory tract showed a short term expression of at least 5 days, but not 10 days, limited only to epithelial cells localised in the epithelium of the nasal mucosa in one out of six calves. Observed limited gene transfer into well differentiated cattle airway epithelial cells must be taken into consideration in order to enhance transfection efficiency, and consequently the vaccine potential of this vector. PMID- 15535962 TI - Mechanism to study 1:1 stoichiometry of NADPH and alkoxyphenoxazones metabolism spectrophotometrically in subcellular biological preparations. AB - Our prior studies have shown that pentoxyresorufin-O-dealkylation (PROD) can be measured spectrophotometrically with simultaneous monitoring of stoichiometry of NADPH/substrate and NADP/product as 10:1:10:1 [Rastogi et al. FEBS Letters 512 (2002) 121-124]. In the present investigation, mechanism of action of other enzymes in modulating the stoichiometry of alkoxyphenoxazones metabolism to 1:1 for electron donor/substrate and oxidized electron donor/product in the same incubation mixture was studied. The spectrophotometric analysis reveals 10:1 ratio between NADPH and pentoxyresorufin (PRF)-ethoxyresorufin (ERF) in microsomal system. The high ratio of electron donor to substrate is due to the presence of the other forms of P-450, which may participate in endogenous metabolism of compounds, thereby reducing the ratio to 4:1 and 7:1 for NADPH/PRF ERF. Incubation of dicumarol in the microsomal PROD or ethoxyresorufin-O dealkylase (EROD) assay led to significant decrease in the consumption of NADPH with a ratio of 4:1 and 7:1 for NADPH/PRF-ERF which is due to inhibition of NADPH cytochrome c (P-450) reductase. In post mitochondrial fraction (S-9), the ratio of 11:1 and 15:1 is seen for NADPH/PRF-ERF. The addition of dicumarol in S-9 fraction showed enhanced rate of alkoxyphenoxazone utilization, suggesting the possibility of reduced resorufin product as a feedback inhibitor. Equating the ratio of NADPH/substrate(s) derived after endogenous utilization of NADPH with the ratio after accounting for NADPH consumption following dicumarol addition in either S-9 or microsomal fraction, a 1:1 mol of NADPH/substrate(s) and oxidized electron donor/product is obtained. The results further suggest that cytosolic fraction may interfere in monitoring the formation of resorufin during dealkylation of alkoxyphenoxazones making dicumarol a mandatory cofactor. PMID- 15535963 TI - Further development of Axisymmetric Drop Shape Analysis-captive bubble for pulmonary surfactant related studies. AB - The methodology combining Axisymmetric Drop Shape Analysis (ADSA) with a captive bubble (ADSA-CB) facilitates pulmonary surfactant related studies. The accuracy of ADSA-CB is crucially dependent on the quality of the bubble profile extracted from the raw image. In a previous paper, an image analysis scheme featuring a Canny edge detector and a Axisymmetric Liquid Fluid Interfaces-Smoothing (ALFI-S) algorithm was developed to process captive bubble images under a variety of conditions, including images with extensive noise and/or lack of contrast. A new version of ADSA-CB based on that image analysis scheme is developed and applied to pulmonary surfactant and pulmonary surfactant-polymer systems. The new version is found to be highly noise-resistant and well self-adjusting. PMID- 15535964 TI - Physicochemical characterization of cross-linked human serum albumin dimer and its synthetic heme hybrid as an oxygen carrier. AB - The recombinant human serum albumin (rHSA) dimer, which was cross-linked by a thiol group of Cys-34 with 1,6-bis(maleimido)hexane, has been physicochemically characterized. Reduction of the inert mixed-disulfide of Cys-34 beforehand improved the efficiency of the cross-linking reaction. The purified dimer showed a double mass and absorption coefficient, but unaltered molar ellipticity, isoelectric point (pI: 4.8) and denaturing temperature (65 degrees C). The concentration dependence of the colloid osmotic pressure (COP) demonstrated that the 8.5 g dL(-1) dimer solution has the same COP with the physiological 5 g dL( 1) rHSA. The antigenic epitopes of the albumin units are preserved after bridging the Cys-34, and the circulation lifetime of the 125I-labeled variant in rat was 18 h. A total of 16 molecules of the tetrakis[(1 methylcyclohexanamido)phenyl]porphinatoiron(II) derivative (FecycP) is incorporated into the hydrophobic cavities of the HSA dimer, giving an albumin heme hybrid in dimeric form. It can reversibly bind and release O2 under physiological conditions (37 degrees C, pH 7.3) like hemoglobin or myoglobin. Magnetic circular dichroism (CD) revealed the formation of an O2-adduct complex and laser flash photolysis experiments showed the three-component kinetics of the O2-recombination reaction. The O2-binding affinity and the O2-association and dissociation rate constants of this synthetic hemoprotein have also been evaluated. PMID- 15535965 TI - Dietary magnesium depletion does not promote oxidative stress but targets apical cells within the mouse caput epididymidis. AB - It is well documented that a dietary deficiency in magnesium can induce oxidative stress and an inflammatory response in animal models. In our study, we have investigated these responses in the mouse epididymis after mice had been fed a magnesium-deficient diet for a 2-week duration. The extracellular and intracellular concentrations of magnesium where shown to be depleted on this diet. This was followed, however, only in the liver of the Mg-deficient animals, by an increase in both alpha 2-macroglobulin (alpha-2m), an acute phase marker, and interleukin-6 transcripts suggesting that an inflammatory response had been initiated. These changes were correlated with a decrease in circulating neutrophils. To address the question of whether or not peroxidation was induced in mouse epididymis following hypomagnesia, we have monitored the level of endogenous peroxidation, their ability to respond to induced peroxidation as well as the expression and activity of the enzymatic glutathione peroxidase (GPX) antioxidant family. To evaluate if the epididymis had evolved specific protections against peroxidation, other organs such as the liver and the kidney were monitored in parallel. We detected no evidence for increased peroxidation in any of the mouse organs tested. However, GPX activity was found to be significantly lower in the liver and the kidney of Mg-deficient animals while it was unchanged in the epididymides of the same animals during the deficiency. Histological analysis of the epididymis showed no major difference in the overall cytological aspect of the organ. Segment 2 of the caput, however presented a significant increase in the number of apically located cells or blebbing cells. Immunohistochemical analysis proved that these cells were epididymal apical cells and not infiltrated leukocytes. These observations suggested that the mouse caput epididymidis segment 2 specifically responded to Mg deficiency via the apical cells. Finally, a comparative analysis of stress response genes was conducted in control and magnesium-deficient caput epididymidis samples. It brought forward some genes that might be involved in the peculiar response of the caput epithelium following hypomagnesia. PMID- 15535966 TI - Reevaluating the role of 1,10-phenanthroline in oxidative reactions involving ferrous ions and DNA damage. AB - It is widely believed that the iron chelator 1,10-phenanthroline (phen) is able to fully block the Fenton reaction by forming a complex (Fe(phen)3(2+), also known as ferroin) that cannot react with H2O2. We observed that phen cannot fully prevent 2-deoxyribose (5 mM) degradation induced by Fenton reagents (30 microM Fe(II) plus 100-500 microM H2O2); protection varied from 55% to 66% when the phen/Fe(II) ratio was 3:1 to 20:1. Inhibition of 2-deoxyribose damage was nearly unchanged if phen was pre-incubated with Fe(II). Moreover, preformed Fe(phen)3(2+) complex added to the solution containing H2O2 was able to induce 2 deoxyribose degradation and methane sulfinic acid formation from the oxidation of 5% DMSO. The partially protective effect of phen was unchanged with the use of either phosphate or HEPES as buffers (5 mM, pH 7.2), or in unbuffered media (pH 5.1). Both DMSO oxidation and 2-deoxyribose degradation correlated with the increase in Fe(phen)3(2+) concentration. Strand breaks in plasmid pTARGETtrade mark DNA induced by Fenton reagents (1 microM Fe(II) plus 25 microM H2O2) in HEPES buffer could only be partially prevented by phen, even when the chelator was 16 times more concentrated than Fe(II). In these experiments, Fe(phen)3(2+) and DNA were pre-incubated from 1 to 10 min before addition of H2O2. Moreover, a high level of DNA strand breakage was observed when iron and phen are added to the reaction immediately before H2O2. On the other hand, phen fully prevented 2 deoxyribose degradation induced by the autoxidation of 30 microM Fe(II) in phosphate-buffered (3 to 30 mM) media. Our data provide evidence that the Fe(phen)3(2+) complex induces in vitro oxidative damage in the presence of H2O2 (possibly by means of Fe(phen)3(2+) dissociation into Fe(phen)2(2+)), but they show that the complex cannot undergo autoxidation. PMID- 15535967 TI - Structural and serological characterization of the major glycolipid from Rothia mucilaginosa. AB - Structural studies on the major glycolipid isolated from Rothia mucilaginosa were carried out utilising specific chemical degradation, NMR spectroscopy and matrix assisted laser-desorption/ionization time of flight mass spectrometry (MALDI TOF MS). The glycolipid was found to be a dimannosylacylmonoglyceride in which the carbohydrate part was the glycerol-linked dimannoside alpha-D-Manp-(1-->3)-alpha D-Manp-(1-->3)-sn-Gro (Man A-Man B-Gro), of which Man B was esterified at O-6 by a fatty acid residue. A second fatty acid substituted the secondary methylene position of the glycerol residue, in contrast to the glycolipid previously found in R. dentocariosa and Saccharopolyspora strains, in which the second fatty acid esterified the primary methylene position of glycerol. Results of the ELISA experiment with rabbit specific antibacterial sera indicate that these two major glycolipids are antigenic, and the patterns of serological reactivity are similar but not identical. PMID- 15535968 TI - Expression of an immunologically reactive merozoite surface protein (MSP-1(42)) in E. coli. AB - The 42-kDa carboxyl-terminal processing fragment of Plasmodium falciparum merozoite surface protein-1 (PfMSP-1(42)) is one of the anti-malarial vaccine candidate antigens. In the present study, recombinant MSP-1(42) was expressed as a fusion protein in a novel E. coli host. The average yield of the recombinant protein was 48 mg/l of bacterial culture. The antigenicity and immunogenicity of the purified protein were evaluated by comparing the results with those obtained from a well-characterized recombinant MSP-1(42) (Bmp42) expressed in the baculovirus expression system previously described from our laboratory. We observed that there is a high degree of similarities between the two recombinant proteins. Based on the results from T and B cell response, in vitro parasite growth inhibition, as well as cross-reactivities with several well-characterized MSP-1 specific Mabs, the bacterial expressed protein is apparently comparable to Bmp42 in terms of immunoreactivities. Our results suggest that the bacterial expression system could be employed to express immunologically active recombinant MSP-1(42) at elevated levels. This system may be an attractive alternative for producing a protective vaccine for human use at lower cost. PMID- 15535969 TI - LacdiNAc (GalNAcbeta1-4GlcNAc) is a major motif in N-glycan structures of the chicken eggshell protein ovocleidin-116. AB - The avian eggshell matrix protein ovocleidin-116 (OC-116) contains two N glycosylation sites in its sequence. One of them, 293N-D-S, is modified only marginally while the second one, 62N-Q-T, is completely occupied by N-linked glycans. The glycopeptide bearing the modified site was isolated by size exclusion chromatography and reversed phase HPLC after cleavage of the protein with lysyl endopeptidase. The carbohydrate structures attached to Asn62 were determined by carbohydrate compositional analysis, methylation analysis and electrospray MS/MS. We identified 17 different oligosaccharide structures. Four of them were of the high-mannose type, eight were hybrid type and five were complex type structures. Both, hybrid and complex type glycans comprised core fucosylated and peripherally fucosylated structures. Most of the antennae contained the relatively rare lacdiNAc (GalNAcbeta1-4GlcNAc) motif, which was fucosylated in 9 out of 15 structures. The lacNAc (Galbeta1-4GlcNAc) motif, which is the more frequent motif in mammals, only occurred in 3 of the 17 glycoforms. This is the first detailed study of N-glycan structures occurring in an avian shell-specific protein and, to our knowledge, the first description of fucosylated lacdiNAc structures present in avian glycoproteins. PMID- 15535970 TI - Assay and subcellular localization of pyrroline-5-carboxylate dehydrogenase in rat liver. AB - Delta1-pyrroline-5-carboxylate dehydrogenase (P5CDh) catalyzes the conversion of Delta1-pyrroline-5-carboxylate to glutamate in a reaction requiring NADP+ as a cofactor. Delta1-pyrroline-5-carboxylate is formed in liver from proline by proline oxidase (EC number not assigned) or from ornithine via ornithine aminotransferase. A spectrophotometric assay for P5CDh was shown to be valid if rotenone was included in the assay to prevent reoxidation of NADH. Using this new assay, liver was fractionated using differential centrifugation and the distribution of P5CDh was compared to that of appropriate marker enzymes. P5CDh is enriched only in the mitochondrial fractions, as are the mitochondrial enzymes, succinate cytochrome c reductase, proline oxidase, glutaminase, and ornithine aminotransferase. Thus, it can be concluded that P5CDh occurs only in mitochondria, not in both mitochondria and cytoplasm, as had previously been reported. PMID- 15535971 TI - A fibronectin fragment induces tumor necrosis factor production of rat basophilic leukemia cells. AB - Proteolytic digest of fibronectin (FN), but not intact FN, induced TNF-alpha secretion of rat basophilic leukemia (RBL-2H3) cells. As a result of the identification of FN fragment responsible for TNF-alpha secretion, a 30-kDa fragment derived from the carboxyl-terminal heparin-binding (Hep 2) domain of FN was isolated from the FN digest. The TNF-alpha secretion was abrogated by treatment of RBL-2H3 cells with cycloheximide, indicating the de novo synthesis of TNF-alpha, but not with polymyxin B, excluding the possible TNF-alpha induction by some contaminated lipopolysaccharides. A 22-mer synthetic peptide originated from the Hep 2 domain, termed FNIII14, which has been found to negatively modulate the beta1 integrin activation, had the ability to induce TNF alpha production, whereas this activity of FNIII14 disappeared by shuffling a YTIYVIAL sequence essential for the integrin-inactivating activity. FNIII14 suppressed the spreading of RBL-2H3 cells on FN substrate, wherein RBL-2H3 cell proliferation was inhibited with FNIII14 in a dose-dependent manner. Thus, it appears that FN fragments containing the YTIYVIAL anti-adhesive site affect the activation status of RBL-2H3 mast cells, characterized by the stimulation of TNF alpha production and growth suppression, probably due to negative regulation of beta1 integrin activity. PMID- 15535972 TI - Deletion of the cytoplasmic domain of human alpha3/4 fucosyltransferase III causes the shift of the enzyme to early Golgi compartments. AB - The transmembrane domain (TM) and flanking regions of glycosyltransferases (GTs) have been implicated in the localization of these proteins in the Golgi apparatus (GA). alpha3/4 Fucosyltransferase III (FT3wt) (EC 2.4.1.65) is localized in the trans-Golgi and trans-Golgi network (TGN) of baby hamster kidney (BHK) cells and synthesizes Lewis determinants associated with cell adhesion events. We have evaluated the effect of removing the cytosolic domain on the localization of the enzyme and its capacity for synthesizing the Lewis A (Le A) determinant. The mutant where the cytoplasmic domain (Asp-2 to Trp-13) of FT3wt has been deleted (FT3dc) was localized in the Golgi but it was shifted to earlier compartments than FT3wt. The mutant was not detected on the plasma membrane (PM) and glycosylation analysis indicated that FT3dc was transported beyond the endoplasmic reticulum (ER) since complex type glycosylation was observed. Cells expressing FT3dc showed a significantly lower efficiency to synthesize Le A when compared with cells expressing FT3wt, in vivo. This reduction was not due to lower specific activity because both enzyme forms had a similar specific activity in vitro. Therefore, removal of FT3 cytosolic tail caused a shift in enzyme distribution to earlier Golgi compartments concomitant to the decrease of its biosynthetic capacity. PMID- 15535973 TI - Hemin/nitrite/H2O2 induces brain homogenate oxidation and nitration: effects of some flavonoids. AB - Oxidative injury has been implicated in the pathogenesis of numerous neurodegenerative diseases. Recently, it has been found that with the existence of hydrogen peroxide and nitrite, hemin catalyzes protein nitration. We hypothesize under certain pathological conditions, hemin catalyzed protein nitration may happen in the brain. In this paper, the effects of three flavonoids, i.e. quercetin, catachin and baicalein on hemin/nitrite/H2O2 induced brain homogenate oxidation and nitration were studied. The results showed that hemin/nitrite/H2O2 system could effectively induce brain homogenate protein oxidation and nitration. Quercetin, catachin and baicalein dose-dependently inhibited hemin/nitrite/H2O2 system-induced protein nitration in a dose-dependent manner, the inhibition of protein nitration was in the order of quercetin>catachin>baicalein. These compounds also inhibited hemin/H2O2 system induced lipid peroxidation, the inhibition order was baicalein >quercetin>catachin. However, these flavonoids showed marginal effect on hemin/nitrite/H2O2 system caused protein oxidation and thiol oxidation. The inhibition activities of flavonoids on hemin/nitrite/H2O2 system-induced protein nitration may closely relate to their radical scavenging activities, since the inhibition order of protein nitration is the same as the radical scavenging order. These results indicate hemin/nitrite/H2O2 system induces different types of oxidative assault on bio-molecules. Flavonoids could act as antioxidants inhibiting ROS and RNS caused brain damage. PMID- 15535974 TI - Fucoxanthin induces apoptosis and enhances the antiproliferative effect of the PPARgamma ligand, troglitazone, on colon cancer cells. AB - The effect of fucoxanthin, from the edible seaweed Undaria pinnatifida on viability of colon cancer cells and induction of apoptosis was investigated. Fucoxanthin remarkably reduced the viability of human colon cancer cell lines, Caco-2, HT-29 and DLD-1. Furthermore, treatment with fucoxanthin induced DNA fragmentation, indicating apoptosis. The DNA fragmentation in Caco-2 cells treated with 22.6 microM fucoxanthin for 24 h was 10-fold higher than in the control. Fucoxanthin suppressed the level of Bcl-2 protein. Also, DNA fragmentation induced by fucoxanthin was partially inhibited by a caspase inhibitor Z-VAD-fmk. Moreover, combined treatment with 3.8 microM fucoxanthin and 10 microM troglitazone, which is a specific ligand for peroxisome proliferator activated receptor (PPAR) gamma, effectively decreased the viability of Caco-2 cells. However, separate treatments with these same concentrations of fucoxanthin nor troglitazone did not affect cell viability. These findings indicate that fucoxanthin may act as a chemopreventive and/or chemotherapeutic carotenoid in colon cancer cells by modulating cell viability in combination with troglitazone. PMID- 15535975 TI - Carboxyl nonsteroidal anti-inflammatory drugs are efficiently glucuronidated by microsomes of the human gastrointestinal tract. AB - Limited studies have been carried out on the biotransformation of carboxyl nonsteroidal anti-inflammatory drugs (NSAIDs) in the liver. However, the role of the intestine in NSAID metabolism has not been investigated. In this report, the contribution of UDP-glucuronosyltransferases (UGTs) in the human gastrointestinal (GI) tract from five donors to the glucuronidation of the NSAIDs, RS-ketoprofen, S-naproxen, RS- and S-etodolac, was investigated. UGT activity and, for some donors, mRNA levels were evaluated. All NSAIDs were glucuronidated throughout the GI tract; however, glucuronidation was low in stomach and duodenum as compared to the remainder of the intestine. RT-PCR analysis demonstrated that the UGT1A isoforms, UGT1A3, 1A8, and 1A10, and UGT2B7 were expressed in the GI tract. Human recombinant UGT1A3, 1A9, 1A10 and 2B7 were actively involved in the glucuronidation of all NSAIDs while UGT1A7 and the intestine-specific UGT1A8 had no glucuronidating activity towards those compounds. Despite interindividual variations in both the levels of mRNA and the distribution of activity through the intestine, UGTs in the GI tract may contribute significantly to the first pass metabolism of orally administered NSAIDs. PMID- 15535976 TI - Salivary thiocyanate/nitrite inhibits hydroxylation of 2-hydroxybenzoic acid induced by hydrogen peroxide/Fe(II) systems under acidic conditions: possibility of thiocyanate/nitrite-dependent scavenging of hydroxyl radical in the stomach. AB - Formation of OH radicals in the stomach is possible by Fenton-type reactions, as gastric juice contains ascorbic acid (AA), iron ions and H2O2. An objective of the present study is to elucidate the effects of salivary SCN- and NO2- on the hydroxylation of salicylic acid which was induced by H2O2/Fe(II) and AA/H2O2/Fe(II) systems. Thiocyanate ion inhibited the hydroxylation of salicylic acid by the above systems in acidic buffer solutions and in acidified saliva. The inhibition by SCN- was deduced to be due to SCN- -dependent scavenging of OH radicals. Nitrite ion could enhance the SCN- -dependent inhibition of the hydroxylation induced by AA/H2O2/Fe(II) systems. The enhancement was suggested to be due to scavenging of OH radicals by NO which was formed by the reactions among AA, HNO2 and SCN- contained in the reaction mixture. The concentrations of SCN- and NO2-, which were effective for the inhibition, were in ranges of their normal salivary concentrations. These results suggest that salivary SCN- can cooperate with NO2- to protect stomach from OH radicals formed by AA/H2O2/Fe(II) systems under acidic conditions. PMID- 15535977 TI - Substrate share in the suicide inactivation of mushroom tyrosinase. AB - To address the real cause of the suicide inactivation of mushroom tyrosinase (MT), under in vitro conditions, cresolase and catecholase reactions of this enzyme were investigated in the presence of three different pairs of substrates, which had been selected for their structural specifications. It was showed that the cresolase activity is more vulnerable to the inactivation. Acetylation of the free tyrosyl residues of MT did not cure susceptibility of the cresolase activity, but clearly decreased the inactivation rate of MT in the presence of 4 [(4-methylbenzo)azo]-1,2-benzenediol (MeBACat) as a catecholase substrate. Considering the results of the previous works and this research, some different possible reasons for the suicide inactivation of MT have been discussed. Accordingly, it was proposed that the interruption in the conformational changes in the tertiary and quaternary structures of MT, triggered by the substrate then mediated by the solvent molecules, might be the real reason for the suicide inactivation of the enzyme. However, minor causes like the toxic effect of the ortho-quinones on the protein body of the enzyme or the oxidation of some free tyrosyl residues on the surface of the enzyme by itself, which could boost the inactivation rate, should not be ignored. PMID- 15535978 TI - Granulin-like peptide in the mid-gut gland of the bivalve mollusk, Patinopecten yessoensis. AB - A cysteine-rich polypeptide, termed CRP1, with a molecular mass of 5829 Da was found to occur in the mid-gut gland of the scallop Patinopecten yessoensis. CRP1 was purified by reverse phase and cation-exchange chromatographies. The amino acid sequence of CRP1 was deduced from its N-terminal amino acid sequence, amino acid composition and the sequence of a partial cDNA, indicating that CRP1 is a 57 amino-acid polypeptide containing 12 cysteine residues with a calculated molecular mass of 5841 Da (5829 Da when oxidized to form six disulfide bridges). A homology search of databases revealed that the deduced amino acid sequence of CRP1 displays significant similarity to those of granulin/epithelins, a family of growth-modulating factors; all cysteine residues in CRP1 are located at the same positions as those conserved characteristically in other known granulin/epithelins. Purified CRP1 inhibited the proliferation of mouse embryo cells. The results suggest that CRP1 functions as a growth-modulating factor in the scallop, and that granulin/epithelin family polypeptides and their precursors play physiologically important roles in invertebrates. PMID- 15535979 TI - Differences in amino acid sequences of mistletoe lectin I and III B-subunits determining carbohydrate binding specificity. AB - Toxic lectins of European mistletoe Viscum album L.--MLI (viscumin), MLII and MLIII--are present in water extracts of this plant. Earlier we have cloned the full-length gene of MLIII precursor [A.G. Tonevitsky, I.I. Agapov, I.B. Pevzner, N.V. Maluchenko, M.M. Mojsenovich, U. Pfueller, M.P. Kirpichnikov, (2004) Biochemistry (Mosc.), 69 (6), 790-800, in press]. Here for the first time we report the cloning and expression in Escherichia coli cells of MLIII gene fragment encoding the carbohydrate-binding subunit. We have proved with our panel of monoclonal antibodies against ML toxins that the cloned fragment encoded MLIII B-subunit. The immunochemical and sugar-binding activities of renatured recombinant MLIII B-subunit were demonstrated in ELISA and ELLA, respectively. The comparative analysis of amino acid sequences of the cloned rMLIIIB and the B subunits of other type II RIPs--MLI, ricin, abrin and nigrin b--was performed, revealing the main differences in primary structure of MLI and MLIII B-chains, which could determine their sugar specificity. The antigenicity analysis of MLI and MLIII B-subunits showed one epitope 25RDDDFRDGNQ34 in MLIB that is absent in MLIIIB sequence. The role of the toxic lectins and their subunits in immunological properties of mistletoe extracts is discussed. PMID- 15535980 TI - Suppression of cell proliferation, DNA protein cross-links, and induction of apoptosis by vanadium in chemical rat mammary carcinogenesis. AB - Vanadium, a dietary micronutrient, has recently been considered as an important pharmacological agent. The present investigation was carried out to ascertain its anticarcinogenic potential against an experimental rat mammary carcinogenesis. Female Sprague-Dawley rats were treated with 7,12dimethylbenz(alpha)anthracene (DMBA) (0.5 mg/100 g body weight) by a single tail vein injection in an oil emulsion. Vanadium (ammonium monovanadate) at a concentration of 0.5 ppm (4.27 micromol/L) was supplemented in drinking water and given ad libitum to the experimental group. The present study was an attempt to assess the effect of vanadium (ammonium monovanadate) on cell proliferation, apoptosis and histopathology in the mammary tissue. We also have examined DNA fragmentation and DNA protein cross-links (DPC) in the liver of rats as well. Immunohistochemical analysis indicated that early neoplasia in mammary tissue proceeds by a decrease in apoptotic cell death (ACD), which was also examined with TUNEL assay, rather than an increase in cell proliferation (P<0.01). DPC in liver were reduced by vanadium treatment (ANOVA, F=13.7, P<0.01). Agarose gel electrophoresis revealed DNA fragmentation in the vanadium-treated group, confirming apoptosis further. Results of the study indicate that the mammary preneoplasia is sensitive to vanadium intervention whereas normal proliferating cells are not. PMID- 15535981 TI - Identification of the 23-kDa peptide derived from the precursor of Gly m Bd 28K, a major soybean allergen, as a new allergen. AB - One of the major soybean allergens, Gly m Bd 28K, is suggested to be biosynthesized as a preproprotein form, which would be composed of a signal peptide, Gly m Bd 28K and the C-terminal peptide (the 23-kDa peptide). However, the 23-kDa peptide has never been characterized. In the present study, we prepared a monoclonal antibody (mAb) against a recombinant 23-kDa peptide expressed in Escherichia coli to detect the 23-kDa peptide in soybean. Several proteins were detected by immunoblotting with the mAb. All of the proteins were shown to have the identical N-terminal amino acid sequence, suggesting that the proteins correspond to the C-terminal part of the Gly m Bd 28K precursor. Furthermore, Gly m Bd 28K and the 23-kDa peptide were observed to come out at the 21st day after flowering and to locate in the crystalloid part of protein storage vacuoles in growing cotyledons. Some of the 23-kDa peptides were shown to be glycoproteins with an N-linked glycan moiety and exhibited the binding to IgE antibodies in the sera of patients sensitive to soybean. The binding of the peptides to IgE antibodies was suggested to be predominantly dependent on their glycan moiety. This study proves the occurrence of the 23-kDa peptide in soybean and that it is a new allergen. PMID- 15535982 TI - Response to acute changes in salinity of two different muscle type creatine kinase isoforms, from euryhaline teleost (Oreochromis mossambicus) gills. AB - Two CKM isoforms (CKM1 and CKM2) from the gills of tilapia (Oreochromis mossambicus) were obtained after transfer from freshwater (FW) to seawater (SW, 25 ppt). Based on the 5' and 3' RACE, the identity of CKM1 and CKM2 was determined to be 59% in the 5'-untranslated region (5'-UTR) and 41.9% in the 3' UTR. Using Northern blot hybridization with the CKM1 and CKM2 3'-UTR probes, CKM1 and CKM2 were found to be expressed in muscle, heart and gill. The levels of these two different CK isoforms (CKM1 and CKM2) were shown to be different in FW than after acute SW transfer, showing that CKM isoforms respond to changes in salinity. PMID- 15535983 TI - Dictyostelium nucleomorphin is a member of the BRCT-domain family of cell cycle checkpoint proteins. AB - A search of the Dictyostelium genome project database (http://dictybase.org/db/cgi-bin/blast.pl) with nucleomorphin, a protein that regulates the nuclear number, predicted it to be encoded by a larger gene containing a putative breast cancer carboxy-terminus domain (BRCT). Using RT-PCR, Northern and Western blotting we have identified a differentially expressed, 2318 bp cDNA encoding a protein isoform of Dictyostelium NumA with an apparent molecular weight of 70 kDa that we have called NumB. It contains a single amino terminal BRCT-domain spanning residues 125-201. Starvation of shaking cultures reduces NumA expression by approximately 88+/-5.6%, whereas NumB expression increases approximately 35+/-3.5% from vegetative levels. NumC, a third isoform that is also expressed during development but not growth, remains to be characterized. These findings suggest NumB may be a member of the BRCT-domain containing cell cycle checkpoint proteins. PMID- 15535984 TI - An evaluation of the P450 inhibition and induction potential of daptomycin in primary human hepatocytes. AB - Two in vitro studies assessed the potential of daptomycin (Cubicin), a newly marketed antibiotic, to affect the cytochrome P450 (CYP450) isoforms in primary cultured human hepatocytes. Both induction and inhibition of isoforms 1A2, 2A6, 2C9, 2C19, 2D6, 2E1, and 3A4 were evaluated. The highest concentrations of daptomycin used in both the induction and inhibition assays were approximately eight-fold higher than the peak total drug concentration (50-60 microg/mL), or the peak free drug concentration (estimated 5-6 microg/mL), in plasma at the clinical dose regimen of 4 mg/kg qd. Results in primary human hepatocytes indicate that daptomycin, at concentrations up to 400 microg total drug/mL, demonstrated no biologically significant induction of any of the CYP450 isoform activities in comparison with the negative control or known inducers. At daptomycin concentrations up to 40 microg free drug/mL, no biologically significant inhibition of the activities of these CYP450 isoforms was observed as compared with known inhibitors. The human hepatocyte results demonstrate that daptomycin has no effects on hepatic CYP450-mediated drug metabolism and, therefore, suggest that daptomycin is unlikely to show potential for pharmacokinetic interactions with concomitantly administered drugs that are metabolized by CYP450 isoforms. PMID- 15535986 TI - Lack of antagonism of 2,3,7,8-tetrachlorodibenzo-p-dioxin's (TCDDs) induction of cytochrome P4501A1 (CYP1A1) by the putative selective aryl hydrocarbon receptor modulator 6-alkyl-1,3,8-trichlorodibenzofuran (6-MCDF) in the mouse hepatoma cell line Hepa-1c1c7. AB - Regulation of gene expression by the aryl hydrocarbon (AHR) receptor is a much studied pathway of molecular toxicology. Activation of AHR by the xenobiotic ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is hypothesized as the mechanism by which TCDD exerts its toxic and carcinogenic effects. Paradoxically, some studies have shown that TCDD acts as an antiestrogen. This has led to the hypothesis that so-called selective aryl hydrocarbon receptor modulators (SAhRMs), AHR ligands that retain the antiestrogenic effects but lack the transcriptional effects of TCDD associated with toxicity, may be utilized as cancer chemotherapeutics in conjunction with other antiestrogenic compounds such as tamoxifen. The present study attempts to further define the molecular mechanism of action of the putative SAhRMs, 6-alkyl-1,3,8-trichlorodibenzofuran (6-MCDF), and diindolylmethane (DIM), focusing particularly on the former. We tested 6-MCDF and DIM for the recruitment of AHR and RNA polymerase II (pol II) to the regulatory region of the AHR responsive gene, cytochrome P4501A1 (CYP1A1), using the chromatin immunoprecipitation (ChIP) assay in the mouse hepatoma cell line Hepa-1c1c7 (Hepa-1). We also tested the level of CYP1A1 induction in Hepa-1 cells using quantitative real-time PCR. We show no difference in the recruitment of AHR or pol II to the regulatory region of CYP1A1 in response to TCDD, 6-MCDF, or co-treatment with both TCDD and 6-MCDF. Our results also show no antagonism of CYP1A1 induction with co-treatment of Hepa-1 cells with TCDD and 6-MCDF. These data suggest that 6-MCDF exhibits agonist activity with respect to induction of CYP1A1 in the Hepa-1 cell line. PMID- 15535985 TI - Functional analysis of human microsomal epoxide hydrolase genetic variants. AB - Human microsomal epoxide hydrolase (EPHX1) is active in the metabolism of many potentially carcinogenic or otherwise genotoxic epoxides, such as those derived from the oxidation of polyaromatic hydrocarbons. EPHX1 is polymorphic and encodes allelic variation at least two amino acid positions, Y113H and H139R. In a number of recent molecular epidemiological investigations, EPHX1 polymorphism has been suggested as a susceptibility factor for several human diseases. To better evaluate the functional contribution of EPHX1 genetic polymorphism, we characterized the enzymatic properties associated with each of the respective variant proteins. Enzymatic profiles were evaluated with cis-stilbene oxide (cSO) and benzo[a]pyrene-4,5-epoxide (BaPO), two prototypical substrates for the hydrolase. In one series of experiments, activities of recombinant EPHX1 proteins were analyzed subsequent to their expression using the pFastbac baculovirus vector in Spodoptera frugiperda-9 (Sf9) insect cells, and purification by column chromatography. In parallel studies, EPHX1 activities were evaluated with human liver microsomes derived from individuals of known EPHX1 genotype. Using the purified protein preparations, rates of cSO and BaPO hydrolysis for the reference protein, Y113/H139, were approximately 2-fold greater than those measured with the other EPHX1 allelic variants. However, when activities were analyzed using human liver microsomal fractions, no major differences were evident in the reaction rates generated among preparations representing the different EPHX1 alleles. Collectively, these results suggest that the structural differences encoded by the Y113H and H139R variant alleles exert only modest impact on EPHX1 specific enzymatic activities in vivo. PMID- 15535987 TI - CNPase activity in myelin from adult rat brains after prolonged lead exposure in vivo. AB - The present study investigated the sensitivity of rats cerebral myelin to prolonged toxicity of lead (Pb) that imitates environmental exposure to this metal. The results indicated that 90 days exposure of young adult rats to lead in drinking water affects the morphology of myelin sheaths, expressed in disintegration of its multilamellar structure. Both, the protein content and the activity of the myelin-specific enzyme CNPase (2',3'-cyclic nucleotide 3 phosphodiesterase), were lowered. The Michaelis-Menten kinetic for CNPase in myelin obtained from control and Pb-treated rats was different. Km increased and Vmax decreased when compared to controls. Observed disturbances in enzyme activity may be one of the potential reasons of the ultrastructural changes. It is thus tempting to speculate that Pb may be considered as a one of the factors contributing to demyelinating diseases. PMID- 15535988 TI - Inhibition of human organic anion transporting polypeptide OATP 1B1 as a mechanism of drug-induced hyperbilirubinemia. AB - OATP1B1 (a.k.a. OATP-C, OATP2, LST-1, or SLC21A6) is a liver-specific organic anion uptake transporter and has been shown to be a higher affinity bilirubin uptake transporter than OATP1B3. Using human embryonic kidney (HEK 293) cells stably transfected with OATP1B1, we have studied the effects of indinavir, saquinavir, cyclosporin A, and rifamycin SV on human OATP1B1 transport function. These drugs are potent inhibitors of OATP1B1 transport activity in vitro. We further provide evidence that the calculated fraction of OATP1B1 inhibited at the clinical exposure level correlated very well with the observed hyperbilirubinemia outcome for these drugs in humans. Our data support the hypothesis that inhibition of OATP1B1 is an important mechanism for drug-induced unconjugated hyperbilirubinemia. Inhibition of OATPs may be an important mechanism in drug drug and drug-endogenous substance interactions. PMID- 15535989 TI - Effects of perfluorocarboxylic acids on the activities of acyl-CoA elongations in vivo and in vitro. AB - Effects of perfluorocarboxylic acids (PFCAs) on proportions of oleic acid and cis vaccenic acid through acyl-CoA chain elongation systems have been studied in the liver of rats. Administration of PFCAs caused a significant increase in palmitoyl CoA chain elongation activity while these chemicals did not affect palmitoleoyl CoA chain elongation activity in vivo. Condensation for both palmitoyl-CoA and palmitoleoyl-CoA were inhibited by PFCAs in vitro at the concentrations, which were physiologically found in the liver of rats treated with the PFCAs. Delta9 Desaturase, which catalyzes both stearoyl-CoA desaturation and palmitoyl-CoA desaturation, was induced by the treatments of rats with the PFCAs. The administration of the PFCAs to rats caused a marked increase in proportion of oleic acid, while that of cis-vaccenic acid was not affected at all. These results strongly suggest that the induced palmitoyl-CoA chain elongation by PFCAs, which exist in the liver, effectively produces oleic acid in concert with the induced stearoyl-CoA desaturase, but the inhibitory effects of PFCAs on either palmitoyl-CoA chain elongation or palmitoleoyl-CoA chain elongation are not crucial for the formation of the elongated fatty acids in vivo. PMID- 15535990 TI - Protective role of zinc in nickel induced hepatotoxicity in rats. AB - This study was planned to determine the protective role of zinc, if any, in attenuating the toxicity induced by nickel sulfate in rat liver. Female Sprague Dawley (SD) rats received either nickel alone in the dose of 800 mg/l in drinking water, zinc alone in the dose of 227 mg/l in drinking water, and nickel plus zinc or drinking water alone for a total duration of eight weeks. The effects of different treatments were studied on various parameters in rat liver which include antioxidant enzymes, levels of nickel and zinc and histoarchitecture at the light microscopic level. Further, the activities of hepatic marker enzymes AST and ALT were also studied in rat serum. Nickel treatment to the normal control animals, resulted in a significant increase in lipid peroxidation and enzyme activities of catalase and glutathione-S-transferase. On the contrary, nickel treatment to normal rats caused a significant inhibition in the levels of reduced glutathione. Superoxide dismutase activity was found to be decreased which however was not significant. Interestingly, when Zn was supplemented to nickel treated rats, the activities of catalase, and glutathione-S-transferase and the levels of GSH and lipid peroxidation came back to within normal limits. Activities of serum AST and ALT were increased significantly following nickel treatment to normal rats. Simultaneous zinc administration to nickel treated rats tended to restore the altered levels of AST and ALT. Normal control and zinc treated animals revealed normal histology of liver. On the other hand, nickel treated animals showed alterations in normal hepatic histoarchitecture which comprise of vacuolization of the hepatocytes and dilatation of sinusoids as well as increase in the number of bi-nucleated cells. Administration of zinc to nickel treated rats resulted in marked improvement in the structure of hepatocytes, thus emphasizing the protective potential of zinc in restoring the altered hepatic histoarchitecture. The nickel administration to normal rats indicated increased concentrations of nickel and decreased concentrations of zinc. However, zinc effectively brought the altered levels of nickel and zinc to within normal range. The study concludes that zinc has the potential in alleviating the toxic effects of nickel in rat liver because of its property to induce metallothionein (S-rich protein) as a free radical scavenger, or its indirect action in reducing the levels of oxygen reactive species. PMID- 15535991 TI - The mouse visual system: from photoreceptors to cortex. Proceedings of a meeting. April 23-24, 2004. Ft. Lauderdale, Florida, USA. PMID- 15535992 TI - From candelas to photoisomerizations in the mouse eye by rhodopsin bleaching in situ and the light-rearing dependence of the major components of the mouse ERG. AB - To quantify the rate at which light in a ganzfeld produces photoisomerizations in mouse rods in situ, we measured the rate of rhodopsin bleaching in eyes of recently euthanized mice with fully dilated pupils. The amount of rhodopsin declined as a first-order (exponential) function of the duration of the exposure at the luminance of 920 scot cd m(-2): the rate constants of bleaching were 8.3 x 10(-6) and 2.8 x 10(-5) s(-1) (scot cd(-1)m2)(-1) for C57B1/6 and 129P3/J mice, respectively. When the approximately 3-fold difference in effective areas of the pupils of the mice are taken into consideration, the bleaching rates for both strains become essentially the same, 2.6 x 10(-6) fraction rhodopsin (scot Td s)( 1). Assuming 7 x 10(7) rhodopsin molecules per rod, this bleaching rate yields the result that a flash of 1 scot Td s produces 181 photoisomerizations per rod, a value close to that derived from analysis of the collecting area of the rod for axially propagating light. We measured the electroretinograms of mice of the two strains reared under controlled illumination conditions (2 and 100 lux), and compared their properties, using the calibrations to determine the absolute sensitivities of the b-wave and a-waves. The intensity that produces a half saturating rod b-wave response is 0.3-0.6 photoisomerizations rod(-1), and the amplification constant of the rod a-wave is 5-6 s(-2) photoisomerization(-1), with little dependence on the strain. PMID- 15535993 TI - In vivo studies of signaling in rod pathways of the mouse using the electroretinogram. AB - PURPOSE: (a) To examine the possibility that there is a threshold in the synaptic mechanism linking rods to rod bipolar cells that can reduce the transmission of continuous noise from the rods without blocking the transmission of any significant proportion of single-photon responses. (b) To estimate the level of this threshold and the amplitude of the continuous noise which it can serve to reduce. (c) To identify the location of the threshold mechanism in the rod to rod bipolar cell pathway. METHODS: Corneal electroretinogram recordings were made from dark-adapted mice anesthetized with ketamine/xylazine after inner-retinal components had been suppressed to isolate PII, the response of depolarizing bipolar cells. Suppression was achieved by intravitreal injections of GABA, TTX, or in Cx36 KO animals by crushing the optic nerve and waiting for ganglion cells to degenerate. RESULTS: All energy-scaled records of isolated PII obtained with ganzfeld stimuli that gave rise to much less than one photoisomerization (R*) per rod (0.01-0.2 R*/rod), had an essentially identical waveform. Stronger stimuli caused a reduction in the peak amplitude of energy-scaled records (saturation) and stimuli strong enough to produce multiple isomerizations in individual rods resulted in a shortening of the response latency and an increase of the energy scaled amplitude at early times (supralinearity). The shape of the rising edge of isolated PII changed with flash energy in a way that was consistent with the existence of a synaptic threshold whose level was less than one tenth of the amplitude of single-photon signals and a continuous noise whose rms amplitude was even less than this. However, when measured at the time of the peak, the amplitude of PII increased linearly in proportion to stimulus energy from the very lowest levels up to the point where there was, on average, 0.2 R*/rod. CONCLUSIONS: There is a threshold nonlinearity operating at the output of the rod to rod bipolar cell synapse that can usefully reduce the transmission of continuous rod noise without significantly affecting the transmission of single photon signals. This nonlinearity does not affect the overall linear function of the rod pathway at levels at which it is effectively operating in a photon counting mode. PMID- 15535994 TI - Transmission of scotopic signals from the rod to rod-bipolar cell in the mammalian retina. AB - Mammals can see at low scotopic light levels where only 1 rod in several thousand transduces a photon. The single photon signal is transmitted to the brain by the ganglion cell, which collects signals from more than 1000 rods to provide enough amplification. If the system were linear, such convergence would increase the neural noise enough to overwhelm the tiny rod signal. Recent studies provide evidence for a threshold nonlinearity in the rod to rod bipolar synapse, which removes much of the background neural noise. We argue that the height of the threshold should be 0.85 times the amplitude of the single photon signal, consistent with the saturation observed for the single photon signal. At this level, the rate of false positive events due to neural noise would be masked by the higher rate of dark thermal events. The evidence presented suggests that this synapse is optimized to transmit the single photon signal at low scotopic light levels. PMID- 15535995 TI - Synaptic circuitry mediating light-evoked signals in dark-adapted mouse retina. AB - Light-evoked excitatory cation current (DeltaIC) and inhibitory chloride current (DeltaICl) of rod and cone bipolar cells and AII amacrine cells (AIIACs) were recorded from slices of dark-adapted mouse retinas, and alpha ganglion cells were recorded from flatmounts of dark-adapted mouse retinas. The cell morphology was revealed by Lucifer yellow fluorescence with a confocal microscope. DeltaIC of all rod depolarizing bipolar cells (DBCRs) exhibited similar high sensitivity to 500 nm light, but two patterns of DeltaICl were observed with slightly different axon morphologies. At least two types of cone depolarizing bipolar cells (DBCCs) were identified: one with axon terminals ramified in 70-85% of IPL depth and DBCR like DeltaIC sensitivity, and the other with axon terminals ramified in 55-75% of IPL depth and much lower DeltaIC sensitivity. The relative rod/cone inputs to DBCs and AIIACs were analyzed by comparing the DeltaIC and DeltaICl thresholds and dynamic ranges with the corresponding values of rods and cones. On average, the sensitivity of a DBCR to the 500 nm light is about 20 times higher than that of a rod. The sensitivity of an AIIAC is more than 1000 times higher than that of a rod, suggesting that AIIAC responses are pooled through a coupled network of about 40 AIIACs. Interactions of rod and cone signals in dark-adapted mouse retinas appear asymmetrical: rod signals spread into the cone system more efficiently than cone signals into the rod system. The mouse synaptic circuitry allows small rod signals to be highly amplified and effectively transmitted to the cone system via rod/cone and AIIAC/DBCC coupling. Three types of alpha ganglion cells (alphaGCs) were identified. (1) ONGCs exhibits no spike activity in darkness, increased spikes in light, sustained inward DeltaIC, sustained outward DeltaICl of varying amplitude, and large soma (20-25 microm in diameter) with an alpha-cell-like dendritic field about 180-350 microm stratifying near 70% of the IPL depth. (2) Transient OFFalphaGCs (tOFFalphaGCs) exhibit no spike activity in darkness, transient increased spikes at light offset, small sustained outward DeltaIC in light, a large transient inward DeltaIC at light offset, a sustained outward DeltaICl, and a morphology similar to the ONalphaGCs except for that their dendrites stratified near 30% of the IPL depth. (3) Sustained OFFalpha GCs (sOFFalphaGCs) exhibit maintained spike activity of 5-10 Hz in darkness, sustained decrease of spikes in light, sustained outward DeltaIC, sustained outward DeltaICl, and a morphology similar to the tOFFalphaGCs. By comparing the response thresholds and dynamic ranges of alphaGCs with those of the pre-ganglion cells, our data suggest that the light responses of each type of alphaGCs are mediated by different sets of bipolar cells and amacrine cells. PMID- 15535996 TI - GABAC receptor-mediated inhibition in the retina. AB - Inhibition at bipolar cell axon terminals regulates excitatory signaling to ganglion cells and is mediated, in part, by GABAC receptors. We investigated GABAC receptor-mediated inhibition using pharmacological approaches and genetically altered mice that lack GABAC receptors. Responses to applied GABA showed distinct time courses in various bipolar cell classes, attributable to different proportions of GABAA and GABAC receptors. The elimination of GABAC receptors in GABAC null mice reduced and shortened GABA-activated currents and light-evoked inhibitory synaptic currents (L-IPSCs) in rod bipolar cells. ERG measurements and recordings from the optic nerve showed that inner retinal function was altered in GABAC null mice. These data suggest that GABAC receptors determine the time course and extent of inhibition at bipolar cell terminals that, in turn, modulates the magnitude of excitatory transmission from bipolar cells to ganglion cells. PMID- 15535997 TI - Function and plasticity of homologous coupling between AII amacrine cells. AB - The AII amacrine cells are critical elements in the primary rod pathway of the mammalian retina, acting as an obligatory conduit of rod signals to both on- and off-center ganglion cells. In addition to the chemical synaptic circuitry they subserve, AII cells form two types of electrical synapses corresponding to gap junctions formed between neighboring AII cells as well as junctions formed between AII cells and on-center cone bipolar cells. Our recent results indicate that coupling between AII cells and cone bipolar cells forms an obligatory synapse for transmission of scotopic visual signals to on-center ganglion cells. In contrast, AII-AII cell coupling acts to maintain the sensitivity of the primary rod pathway by allowing for summation of synchronous activity and the attenuation of asynchronous background noise. Further, the conductance of AII-AII cell gap junctions is highly dynamic, regulated by ambient light conditions, thereby preserving the fidelity of rod signaling over the scotopic operating range from starlight to twilight. PMID- 15535998 TI - Visual experience and maturation of retinal synaptic pathways. AB - The retinal synaptic network continues its maturational refinement after eye opening in mammals. This synaptic refinement is reflected in changes of retinal neuron synaptic activity and connectivity. In mature retina, the dendrites of retinal ganglion cells (RGCs) in the inner plexiform layer (IPL) of retina are separated into ON or OFF sublamina. At early developmental stage, however, the dendrites of most RGCs are ramified throughout the IPL. Recently we found that the postnatal maturational processes converting bistratified ON-OFF responsive RGCs to monostratified ON and OFF responsive RGCs depend upon visual stimulation after eye opening. PMID- 15535999 TI - Localization of zinc transporter-3 (ZnT-3) in mouse retina. AB - Studies of the central nervous system have localized the zinc-transporter-3 (ZnT 3) protein to synaptic vesicles containing glutamate and zinc. We have examined the distribution of the ZnT-3 protein in the light-adapted mouse retina using immunohistochemical techniques. Light microscopic analysis of 15-30-microm retinal sections revealed a rich band of ZnT-3 protein in the region of the outer limiting membrane and photoreceptor inner segments. ZnT-3 reactivity was also present in the outer plexiform, inner nuclear, inner plexiform, and ganglion cell layers. The outer nuclear layer and photoreceptor outer segments did not exhibit ZnT-3 immunoreactivity. In the light-adapted murine retina, ZnT-3 appears localized in regions which have been found reactive for ionic zinc. PMID- 15536000 TI - Developmental defects in Rb-deficient retinae. AB - We recently found that the Rb protein is important for the regulation of retinal progenitor cell proliferation and rod photoreceptor development in the mouse retina. These two functions are separate for Rb and in this study we further characterize the role of Rb in retinal development. At postnatal day 12 in the retinae of Chx10-Cre;RbLox/- mice, immature cells are found in the outer nuclear layer where rods normally are differentiating. This results in alternating patches of the outer nuclear layer (ONL) that are lacking rod inputs. At this stage of development, horizontal cell processes at the outer plexiform layer do not mature appropriately and they extend into the outer nuclear layer. These disruptions in horizontal cell differentiation can persist for several weeks into the adult stage. While there are several secondary effects of the loss of Rb on retinal development including, limited cell death in the ONL, Muller glial cell activation, persistence of immature cells in the ONL, and altered nuclear morphology of cells in the ONL, we suggest that the defect in horizontal cell synapse formation at the OPL results from fewer rod inputs. Mice with other developmental defects in photoreceptor cell fate specification or glial cell activation do not exhibit a similar alteration in horizontal cell differentiation. Therefore, the retinae from Chx10-Cre;RbLox/- mice represent a unique model to study the role of rod photoreceptor inputs in horizontal cell differentiation and synapse formation. PMID- 15536002 TI - L-type calcium channel agonist induces correlated depolarizations in mice lacking the beta2 subunit nAChRs. AB - Retinal waves are mediated in part by activation of nicotinic receptors containing the beta2 subunit. Mice deficient in beta2 containing nAChRs have maintained firing of action potentials but do not support correlated waves. As a result, beta2-/- mice have inhibited refinement of circuits within the retina as well as retinal projections to the CNS. Previously, we observed that correlated increases in calcium reminiscent of retinal waves could be induced in beta2-/- retina by pharmacological application of the L-type calcium channel agonist, FPL 64176. Here, we characterize FPL-induced activity patterns in beta2-/- retina using both whole cell and multielectrode array recordings. FPL-induced strong depolarizations in previously non-spiking beta2-/- retinal ganglion cells. Though these strong depolarizations were likely to underlie the FPL-induced calcium transients, they led to highly variable effects on the spiking of individual retinal ganglion cells. In addition, induced spiking activity had significantly weaker nearest-neighbor correlations than WT mice. Initial attempts of intraocular injections of FPL in beta2-/- mice did not rescue eye-specific layer formation. These findings indicate that activity induced by FPL is not sufficient for driving eye-specific segregation in beta2-/- mice. PMID- 15536001 TI - Results from screening over 9000 mutation-bearing mice for defects in the electroretinogram and appearance of the fundus. AB - Random mutagenesis combined with phenotypic screening using carefully crafted functional tests has successfully led to the discovery of genes that are essential for a number of functions. This approach does not require prior knowledge of the identity of the genes that are involved and is a way to ascribe function to the nearly 6000 genes for which knowledge of the DNA sequence has been inadequate to determine the function of the gene product. In an effort to identify genes involved in the visual system via this approach, we have tested over 9000 first and third generation offspring of mice treated with the mutagen N ethyl-N-nitrosourea (ENU) for visual defects, as evidenced by abnormalities in the electroretinogram and appearance of the fundus. We identified 61 putative mutations with this procedure and outline the steps needed to identify the affected genes. PMID- 15536003 TI - Adenylate Cyclase 1 dependent refinement of retinotopic maps in the mouse. AB - Development of the retino-collicular pathway has served as an important model system for examining the cellular mechanisms responsible for the establishment of neuronal maps of the sensory periphery. A consensus has emerged that molecular or chemical cues are responsible for the initial establishment of gross topography in this map, and that activity dependent factors sharpen this initial rough topography into precision. However, there is little evidence available concerning the biochemical signaling mechanisms that are responsible for topographic map refinement in the retino-collicular system. Using a combination of anatomical and biochemical techniques in normal and mutant mice, we provide evidence that Ca2+/Calmodulin regulated Adenylate Cyclase 1 (AC1), which is strongly expressed in the superficial layers of the colliculus, is an important downstream signaling agent for activity dependent map refinement in the superior colliculus. PMID- 15536004 TI - Biochemical and anatomical subdivision of the dorsal lateral geniculate nucleus in normal mice and in mice lacking the beta2 subunit of the nicotinic acetylcholine receptor. AB - The cytoarchitectonically-uniform dorsal lateral geniculate nucleus (dLGN) can be biochemically and anatomically subdivided in wild-type mice: The nucleus' dorsolateral 'shell' region contains the majority of cells positive for the calcium-binding protein calbindin-D28k, and receives the strongest concentration of inputs from the superior colliculus. This subdivision remains normal in mice lacking the beta2 subunit of the nicotinic acetylcholine receptor. Although in these animals the dLGN contains fewer calbindin-positive cells, those cells are predominantly situated in the dorsolateral portion of the nucleus, and this region remains preferentially targeted by the colliculogeniculate projection. PMID- 15536005 TI - LTP and LTD vary with layer in rodent visual cortex. AB - Mechanisms of plasticity in the visual cortex have been studied with long-term potentiation (LTP), long-term depression (LTD) and ocular dominance plasticity (ODP). It is now possible to compare results from these three forms of plasticity using knockout mice, and also by pharmacological manipulations. A review of the literature shows that if both LTP and LTD are completely abolished, then ODP will also be abolished. In other situations, there is little correlation. We hypothesize that this lack of correlation occurs because the mechanisms for LTP and LTD vary with layer in the visual cortex, and results show that they do. PMID- 15536006 TI - Swept contrast visual evoked potentials and their plasticity following monocular deprivation in mice. AB - The swept contrast visual evoked potential technique is a quasi-psychophysical method that can help bridge the gap between cell biology and visual performance in studies of ocular dominance plasticity. In mice we found that four days of monocular deprivation diminished the amplitude of evoked potentials from the deprived eye relative to the non-deprived eye. This ocular dominance plasticity was nearly as great in adult mice as in juveniles. The monocular deprivation effect was mediated, at least in part, by enhancement of responses evoked from the non-deprived eye, rather than by depression of responses from the deprived eye. PMID- 15536007 TI - Experience-dependent development of feedforward and feedback circuits between lower and higher areas of mouse visual cortex. AB - Using whole cell recordings, we studied excitatory and inhibitory postsynaptic currents (EPSCs, IPSCs) in feedforward (FF) and feedback (FB) circuits between areas V1 and LM (lateromedial) in developing mouse visual cortex. We found that in mice reared with normal visual experience, FF and FB synapses onto layer 2/3 pyramidal neurons develop equal but submaximal strengths whose EPSCs are increased by monocular lid suture. In contrast, the development and experience dependence of FF- and FB-IPSCs is pathway-specific. The difference develops during the critical period by strengthening FF-IPSCs, while keeping FB-IPSC amplitudes constant. Monocular lid suture increases FB-IPSCs but does not affect FF-IPSCs. PMID- 15536008 TI - Using eye movements to assess brain function in mice. AB - Examining eye movements is an important part of the neurological evaluation of humans; the distribution of the neural circuits that control these movements is such that they are disrupted--often in highly characteristic fashions--by many disease processes. Technical advances have made it possible to measure accurately the eye movements of mice, so it is now possible to use the detective power of eye movement recording to characterize neurological dysfunction in genetically altered strains. Here we introduce analytical tools used in ocular motor research and demonstrate their ability to reveal disorders of the visual pathways, inner ear, and cerebellum. PMID- 15536009 TI - Characterization of mouse cortical spatial vision. AB - Little is known about the spatial vision of mice or of the role the visual cortex plays in mouse visual perception. In order to provide baseline information upon which to evaluate the spatial vision of experimentally and genetically altered mice, we used the visual water task to assess the contrast sensitivity and grating acuity of normal C57BL/6 mice. We then ablated striate cortex (V1) bilaterally and re-measured the same visual functions. Intact mice displayed an inverse "U"-shaped contrast sensitivity curve with a maximum sensitivity near 0.2 cycles/degree (c/d). Grating acuity, measured either by discriminating a sine wave grating from an equiluminant gray, or vertical from horizontal sine wave gratings, was near 0.55 c/d. Grating acuity and contrast sensitivity were reduced significantly following aspiration of V1. The mouse visual system exhibits fundamental mammalian characteristics, including the feature that striate cortex is involved in processing visual information with the highest sensitivity and spatial frequency. PMID- 15536010 TI - Comparison of plasticity and development of mouse optokinetic and vestibulo ocular reflexes suggests differential gain control mechanisms. AB - Image stability during self-motion is achieved via a combination of the optokinetic and vestibulo-ocular reflexes (OKR and VOR). To determine whether distinct neuronal mechanisms are used to calibrate eye movements driven by visual and vestibular signals, we examined the developmental maturation and adaptive plasticity of the OKR and VOR in mice. The combined performance of the OKR and VOR, measured with infrared video oculography, produces nearly perfect gaze stability both in adult mice and in juveniles (postnatal days 21-26). Analyses of the OKR and VOR in isolation, however, indicate that VOR gains in juveniles are lower than in adult mice, while OKR gains are higher, indicating that juveniles rely more strongly on vision to stabilize gaze than do adults. Adaptive plasticity in the mouse OKR and VOR could be induced by 30 min of visual vestibular mismatch training. Examination of the effects of training on the OKR and VOR revealed differential mechanisms and persistence of adaptive plasticity. Increases in VOR gain induced by rotating mice in the opposite direction to the visual surround were short-lasting and were accompanied by long-lasting increases in OKR gain. In contrast, decreases in VOR gain induced by rotating mice in the same direction as the visual surround were persistent and were accompanied by long-lasting increases in OKR gain. Vestibular training had little effect on either the OKR or VOR, while visual training induced robust and long-lasting increases in the OKR but had no effect on the VOR. These data indicate that multiple mechanisms of plasticity operate over distinct time courses to optimize oculomotor performance in mice. PMID- 15536012 TI - Early events in the DNA damage response. AB - The ability to sense DNA damage and activate response pathways that coordinate cell cycle progression and DNA repair is essential for the maintenance of genomic integrity and the viability of organisms. During the last couple of years, several proteins have been identified that participate very early in the DNA damage response. Here we review the current understanding of the mechanisms by which mammalian cells detect DNA lesions, especially double-strand breaks, and mediate the signal to downstream transducers. PMID- 15536013 TI - Afrotherian origins and interrelationships: new views and future prospects. PMID- 15536014 TI - The role of antisense transcription in the regulation of X-inactivation. PMID- 15536015 TI - The genetics of hiding the corpse: engulfment and degradation of apoptotic cells in C. elegans and D. melanogaster. PMID- 15536016 TI - Beginning and ending an actin filament: control at the barbed end. AB - Dynamic actin filaments contribute to cell migration, organelle movements, memory, and gene regulation. These dynamic processes are often regulated by extracellular and?or cell cycle signals. Regulation targets, not actin itself, but the factors that determine it's dynamic properties. Thus, filament nucleation, rate and duration of elongation, and depolymerization are each controlled with regard to time and?or space. Two mechanisms exist for nucleating filaments de novo, the Arp23 complex and the formins; multiple pathways regulate each. A new filament elongates rapidly but transiently before its barbed end is capped. Rapid capping allows the cell to maintain fine temporal and spatial control over F-actin distribution. Modulation of capping protein activity and its access to barbed ends is emerging as a site of local regulation. Finally, to maintain a steady state filaments must depolymerize. Depolymerization can limit the rate of new filament nucleation and elongation. The activity of ADF?cofilin, which facilitates depolymerization, is also regulated by multiple inputs. This chapter describes (1) mechanism and regulation of new filament formation, (2) mechanism of enhancing elongation at barbed ends, (3) capping proteins and their regulators, and (4) recycling of actin monomers from filamentous actin (F-actin) back to globular actin (G-actin). PMID- 15536017 TI - Life extension in the dwarf mouse. AB - Ames dwarf mice and Snell dwarf mice lack growth hormone (GH), prolactin (PRL), and thyroid-stimulating hormone (TSH), live much longer than their normal siblings, and exhibit many symptoms of delayed aging. "Laron dwarf mice," produced by targeted disruption of the GH receptor/GH-binding protein gene (GHR KO mice), are GH resistant and also live much longer than normal animals from the same line. Isolated GH deficiency in "little" mice is similarly associated with increased life span, provided that obesity is prevented by reducing fat content in the diet. Long-lived dwarf mice share many phenotypic characteristics with genetically normal (wild-type) animals subjected to prolonged caloric restriction (CR) but are not CR mimetics. We propose that mechanisms linking GH deficiency and GH resistance with delayed aging include reduced hepatic synthesis of insulin like growth factor 1 (IGF-1), reduced secretion of insulin, increased hepatic sensitivity to insulin actions, reduced plasma glucose, reduced generation of reactive oxygen species, improved antioxidant defenses, increased resistance to oxidative stress, and reduced oxidative damage. The possible role of hypothyroidism, reduced body temperature, reduced adult body size, delayed puberty, and reduced fecundity in producing the long-lived phenotype of dwarf mice remains to be evaluated. An important role of IGF-1 and insulin in the control of mammalian longevity is consistent with the well-documented actions of homologous signaling pathways in invertebrates. PMID- 15536018 TI - Quantitative measurement of retinal thickness in patients with diabetic macular edema is useful for evaluation of therapeutic agents. AB - The effect of lisinopril (an angiotensin-converting enzyme inhibitor) on diabetic macular edema (DME) was investigated by quantitative measurement of macular thickness. In a nonrandomized clinical trial, 19 normotensive type 2 diabetic patients with DME prospectively received oral lisinopril therapy for 2 months. Another 10 normotensive type 2 diabetic patients with similar DME were prospectively followed for two months without treatment. Central macular thickness was measured with a retinal thickness analyzer (RTA). In the lisinopril group, visual acuity improved by two lines or more in two out of 19 eyes (11%), was unchanged in 15 eyes (78%), and deteriorated by two lines or more in two eyes (11%). The mean central macular thickness was significantly reduced after 2 months of treatment (381.3 +/- 121.1 microm) compared with that before administration (475.2 +/- 171.0 microm, P = 0.0093). In the control group, central macular thickness was not significantly decreased after 2 months (458.5 +/- 113.7 microm, P = 0.2178) compared with the baseline value (464.7 +/- 152.2). Fluorescein angiography showed that macular leakage was decreased in 10 patients from the lisinopril group (53%) and was unchanged in nine patients (47%). There was a significant difference of central macular thickness between the patients with and without improvement of macular leakage (P = 0.0040). Lisinopril therapy may reduce macular thickness in patients with DME, as shown by this quantitative study. In addition, quantitative measurement of retinal thickness is useful when evaluating therapeutic agents for DME. PMID- 15536019 TI - LDL particle size and lipid composition are risk factors for microalbuminuria in normotensive and normocholesterolemic patients with type 2 diabetes. AB - The study was to evaluate the influence of particle size and lipid composition of low-density lipoprotein (LDL) on urinary albumin excretion and oxidative susceptibility of LDL, and to define association between LDL particle size and alpha-tocopherol content in LDL from normotensive and normocholesterolemic patients with type 2 diabetes. Twenty-three patients with type 2 diabetes (13 males, 10 females) were studied, and none of these patients had hypertension, hypercholesterolemia and overt proteinuria. The baseline body mass index of all patients was less than 28 kg/m2. All patients were hospitalized in Hirosaki University Hospital and took dietary therapy whose total intake was restricted to less than 30 kcal/kg of ideal body weight for 3 weeks. Their plasma glucose levels were controlled within fasting plasma glucose <140 mg/dl and 2-h postprandial plasma glucose <200 mg/dl. LDL particle size was evaluated by using high-resolution polyacrylamide gel electrophoresis (Lipoprint LDL System) and expressed by Rf value. LDL was incubated with 0.25 microM CuSO4 for 20 h, and the degree of LDL oxidation was determined by malondialdehide analysis. Twenty-four hour urinary C-peptide excretion and plasma triglyceride concentration in patients with microalbuminuria were significantly higher than those in normoalbuminuric patients. Rf values in microalbuminuric patients were significantly greater than those in normoalbuminuric patients. There were significantly inverse correlations between Rf value and alpha-tocopherol content in LDL, and between Rf value and LDL-free cholesterol/LDL-total cholesterol. Thiobarbituarte-reactive substance level in LDL had a tendency to correlate with Rf value and significantly inverse correlation to alpha-tocopherol content in LDL. In type 2 diabetics without hypertension, hypercholesterolemia and obvious obesity, smaller LDL particle size, accompanied by mild hyperinsulinemia and mild hypertriglyceridemia seems to be one of the important factors responsible for microalbuminuria. In addition, the present study suggests that the decrease of alpha-tocopherol content in small LDL particle is associated with oxidative susceptibility to Cu2+-induced oxidation. PMID- 15536020 TI - Daily stress and glycaemic control in Type 1 diabetes: individual differences in magnitude, direction, and timing of stress-reactivity. AB - The aim of this study was to investigate the relationships between daily stress and glycaemic control in 54 people with Type 1 diabetes over 21 days. Measures included daily reports of stress (hassles), four-times-daily blood glucose measurements, and HbA1c levels. Time-series analyses revealed considerable variation between individuals in the nature and extent of blood glucose response to stress (stress-reactivity). In approximately one-third of the sample, stress was significantly associated with either same- or next-day blood glucose levels (r-range: -0.79 to 0.58). The majority of stress-reactive individuals (20.4% of the sample) demonstrated a positive association between hassles and same-day blood glucose levels. A much less common effect was found in two individuals (3.7%), where hassles were related to decreased same-day blood glucose. 'Stress reactive' individuals tended to have high HbA1c values at baseline (t(52) = 2.2; P < 0.05), and significant relationships between emotion-focused coping and blood glucose levels (r = 0.93; P < 0.01). In conclusion, although a significant majority of this sample was resistant to the effects of stress, marked individual differences were found in the nature and extent of stress-reactivity. Our study goes beyond other published results as it is longitudinal, uses time-series analyses and includes a relatively larger sample. Clinicians need to be aware of these individual differences in order to advise patients about anticipating and preventing stress-related disruptions of glycaemic control. PMID- 15536021 TI - Association between LDL particle size and postprandial increase of remnant-like particles in Japanese type 2 diabetic patients. AB - Small, dense LDL, as well as chylomicron- and VLDL-remnant lipoproteins, are known to be important risk factors for coronary heart disease in patients with type 2 diabetes mellitus. The aim of this study was to clarify the relationship between LDL particle size and postprandial remnant lipoprotein levels in Japanese type 2 diabetic patients. Forty-six patients with type 2 diabetes mellitus were divided into tertiles according to LDL particle size. The peak LDL particle diameter was <26.30 nm in tertile 1, 26.30-26.85 nm in tertile 2, and >26.85 nm in tertile 3. After a test meal, tertile 1 had a significantly greater increment of triglycerides (TG), remnant-like particle (RLP)-TG, and RLP-cholesterol (RLP C) than tertiles 2 and 3. There was a negative correlation between LDL particle size and the postprandial increases of TG, RLP-TG, and RLP-C. These results indicate that smaller sized LDL particles may be a marker of fasting state for an exaggerated postprandial increase of remnant lipoproteins as well as an increase of TG-rich lipoproteins. PMID- 15536022 TI - The insulin sensitivity, glucose effectiveness and acute insulin response to glucose load of non-obese adolescent type 2 diabetes. AB - To determine the clinical characteristics in adolescent type 2 diabetes (young diabetes, YDM) in Taiwan, we enrolled 11 males who were diagnosed with YDM before 19 years of age into our study. Another 11 patients with adult type 2 diabetes (mature age diabetes, MADM) who were diagnosed after the age of 40 were enrolled as compare group. Subjects from both groups were being treated with oral hypoglycemic agents only at the time of enrollment, and none of the subjects had a history of diabetic ketoacidosis. Plasma lipid levels were measured from the fasting plasma sample. A homeostasis model assessment was used to estimate insulin sensitivity (HOMA-S) and beta-cell function (HOMA-B). Frequent-sampled intravenous glucose tolerance test was also performed to measure the insulin sensitivity (S(I)), glucose effectiveness (E(G)), and acute insulin response after glucose load (AIR). After adjusting for age and BMI, fasting plasma glucose, total cholesterol, HDL-cholesterol, triglycerides, and HOMA-B levels were similar between two groups. The fasting plasma insulin and HOMA-S were significantly higher in YDM. However, the S(I), E(G) and AIR in both groups were also not significantly different. In conclusion, the early onset of diabetes in YDM may be due to the early deterioration of the S(I), E(G) and AIR with similar severity compared with MADM. The role of E(G) might be more important than previously thought in these patients. Finally, the YDM might be a subtype of type 2 diabetes. PMID- 15536023 TI - Treatment satisfaction after commencement of insulin in Type 2 diabetes. AB - Fifty patients with Type 2 diabetes were recruited for a study assessing treatment satisfaction and well-being, following commencement on insulin during routine clinical care. A month after starting insulin, participants completed a Diabetes Treatment Satisfaction Questionnaire (DTSQ) and a Well-being Questionnaire. At seven months, the same questionnaires and clinical measurements were repeated and, in addition, patients completed a Diabetes Treatment Satisfaction Questionnaire (change) (DTSQc). Mean HbA1c decreased from 9.69 to 8.26% (P < 0.001), and the lipid profile showed a corresponding improvement. BMI increased from 30.9 to 32.0 kgm(-2) (P < 0.001). Perceived frequency of hyperglycaemia decreased (P = 0.021), correlating with the change in HbA1c (r = 0.399, P = 0.006). The DTSQc showed improvements at seven months in scores for treatment satisfaction, convenience, flexibility, understanding of diabetes, willingness to recommend the treatment to others, and satisfaction to continue current treatment (all P < 0.001). There was no relationship between weight gain, HbA1c and total or subscale scores. In summary, after seven months of insulin treatment, patients reported positive changes in their perception of treatment satisfaction. These findings, in conjunction with the known health benefits of improved glycaemic control, should encourage practitioners to consider insulin early in the management of Type 2 diabetes. PMID- 15536024 TI - Ankle brachial pressure index and carotid intima-media thickness as atherosclerosis markers in Japanese diabetics. AB - The aim of this study was to assess the clinical significance of ankle brachial pressure index (ABI) and carotid intima-media thickness (IMT) in Japanese patients with type 2 diabetes. ABI and ultrasonographic carotid IMT measurements were made in 1311 patients and the relationships between ABI, IMT, and cardiovascular diseases were examined. Patients were assigned to one of three groups depending on their ABI index: (i) ABI > or = 1.0; (ii) ABI from > or = 0.9 to < 1.0; and (iii) ABI < 0.9. The odds ratios (ORs) for groups (ii) and (iii) compared with (i) for the prevalence of coronary heart disease (CHD), cerebrovascular disorder (CVD), and carotid atherosclerosis (mean carotid IMT > or = 1.1mm) demonstrated that these conditions were inversely related to ABI. As the combined ABI/carotid IMT score increased, each OR for the prevalence of CHD and CVD increased significantly. This suggests that a lower ABI is associated with generalized atherosclerosis. Measurements of the ABI and carotid IMT might provide a good prognostic indicator, and both should be assessed during screening for atherosclerosis in Japanese patients with type 2 diabetes. PMID- 15536025 TI - Correlates of satisfaction for the relationship with their physician in type 2 diabetic patients. AB - In the context of an Italian nation-wide outcomes research program on type 2 diabetes, we investigated the contribution of both patient and setting-related factors to patient satisfaction with their relationship with their physicians. The level of patient satisfaction was measured using the American Board of Internal Medicine (ABIM) 14 patient satisfaction questionnaire. The main results were obtained using multilevel analysis, a statistical technique that takes into account the clustered nature of our data. Overall, 3563 patients were recruited by 101 diabetologists and 103 general practitioners (GPs). Information on patients' satisfaction was available for 2515 patients (71% of the whole sample). Patients' satisfaction was related to patient characteristics and attitudes, but not with physician's sex, age, speciality, and setting of care. In particular, patients who were less likely to delegate to physicians responsibility for diabetes management and those perceiving a lower degree of involvement in disease management showed lower levels of satisfaction. Lower satisfaction scores were also related to lower levels of school education, more severe clinical conditions, and lower psychological adaptation to diabetes. However, patients reporting higher levels of diabetes related worries and more frequent encounters with health care providers showed higher levels of satisfaction. In conclusion, patient satisfaction with physicians' humanness and communication skills is strongly related to personal characteristics, attitudes, expectations, and perceived health. In deciding the best decision-making approach to adopt in individual patients, it is of primary importance to measure how the patient perceives and engages in relationships. PMID- 15536026 TI - Is parity associated with earlier diagnosis of type 2 diabetes? AB - AIM: Whether parity promotes the development of diabetes has been a subject of controversy. Earlier population studies have produced discrepant findings. The aim of the current study was to investigate this question by examining the relationship between parity and the age of diagnosis of diabetes. METHODS: A retrospective analysis from a diabetes database was conducted. Data for 2102 women with type 2 diabetes (T2DM), including date of birth, age at diagnosis, parity, body mass index (BMI), and family history were extracted. A multivariate linear regression model was applied to ascertain if there was an association between parity and the age of diagnosis of diabetes. RESULTS: With correction for the year of birth, there was a significant association between parity and the age of diagnosis of diabetes (beta = -0.25 +/- 0.08, P = 0.002, where beta is the regression coefficient). That is, for an increase in parity by one, the age of diagnosis decreased by 0.25 years. When further adjustments were made for the other variables in the data set, the effect on the overall cohort disappeared, but it persisted in subjects born before 1950 (beta = -0.17 +/- 0.1, P = 0.09), or with a parity >5 (beta = -0.60 +/- 0.25, P = 0.02). CONCLUSIONS: There is an association between parity and the age of diagnosis of diabetes, suggesting that pregnancy may promote the development of diabetes. However, the magnitude of the effect is small, and unlikely to be of great clinical significance in society today. PMID- 15536027 TI - The burden of diabetes and impaired fasting glucose in India using the ADA 1997 criteria: prevalence of diabetes in India study (PODIS). AB - This random multistage cross sectional population survey was undertaken to determine the prevalence of diabetes mellitus (DM) and impaired fasting glycemia/glucose (IFG) in subjects aged 25 years and above in India. The study was carried out in 108 centers (49 urban and 59 rural) to reflect the size and heterogeneity of the Indian population. 41,270 (20,534 males and 20,736 females) subjects were studied. 21,516 (10,865 males and 10,651 females) were from urban areas and 19,754 (9669 males and 10,085 females) from rural areas. Blood samples were taken after a fast of 10-12h and the subjects were categorized as having IFG or DM using the 1997 American Diabetes Association criteria. The age and gender standardized prevalence rate for DM and IFG in the total Indian population was 3.3 and 3.6% respectively (P < 0.001). The standardized prevalence of DM and IFG in urban areas was significantly higher than that for the rural population (urban DM prevalence 4.6% versus rural DM prevalence 1.9%, P < 0.001; urban IFG prevalence 4.8% versus rural IFG prevalence 2.5%, P < 0.001). There was no statistically significant difference in the prevalence between DM (4.6%) and IFG (4.8%) in the urban population. The rural prevalence of IFG (2.5%) was significantly (P <0.001) more than the rural prevalence of DM (1.9%). Type 2 diabetes is a major health problem is India. PMID- 15536028 TI - Comparing the ADA 1997 and the WHO 1999 criteria: Prevalence of Diabetes in India Study. AB - This random multistage cross-sectional population survey was undertaken to determine the prevalence of type 2 diabetes mellitus (DM) in subjects aged 25 years and above in India. The study was carried out in 77 centres (42 urban and 35 rural) to reflect the size and heterogeneity of the Indian population. 18,363 (9008 male and 9355 female) subjects were studied. 10,617 (5379 males and 5238 females) were from urban areas and 7746 (3629 males and 4117 females) from rural areas. Blood samples were taken after a fast of 10-12 and 2 h after 75 g of oral glucose. Subjects were categorized as having impaired fasting glycemia (IFG) or DM using the 1997 ADA or having impaired glucose tolerance (IGT) or DM using the 1999 WHO criteria. The age- and gender-standardized prevalence rate for DM using the ADA criteria was 3.6% whilst that using the WHO criteria was 4.3% (P < 0.001). The respective standardized prevalence of DM, using the two criteria was, 4.7 and 5.6%, respectively (P < 0.001) in the urban Indian population and 2.0 and 2.7% (P < 0.02) in the rural Indian population. Using the WHO criteria, 581 subjects were newly diagnosed whilst the ADA criteria newly diagnosed 437 subjects. The respective numbers for the urban population were 425 and 323, and for the rural population were 146 and 114, respectively. The ADA criteria could diagnose 75.2, 76.0 and 73.0% of the subjects who had DM as per the WHO criteria. Of 739 Indian subjects who had IFG, 106 (14.3%) were diagnosed as having DM by the WHO criteria whilst 505 (68.3%) had values compatible with a diagnosis of IGT. Of the 536 urban subjects with IFG, 74 (13.8%) had DM and 350 (65.3%) had IGT using the WHO criteria. Of the 302 rural subjects with IFG, 32 (15.8%) had DM and 155 (76.3%) had IGT using the WHO criteria. 505 (49.9%) of 1012 Indian subjects with IGT as per the WHO criteria had IFG. 350 (47.7%) of 733 urban subjects and 155 (55.5%) of 279 rural subjects with IGT had values compatible with IFG as per the ADA criteria. Type 2 diabetes is a major health problem is India. The use of the ADA criteria would underestimate the prevalence of DM by not diagnosing subjects showing a poor response to a glucose challenge. This along with the discrepancies between subjects showing IGF or IGT could be a challenge to any prevention program. PMID- 15536029 TI - Synthesis of standing-up trajectories using dynamic optimization. AB - Dynamic optimization as a tool to compute standing-up trajectories was investigated. Sit-to-stand manoeuvres in five intact persons and five trans femoral amputees were measured. Movements and ground reaction forces acting on the body were recorded. A five-segment 3D dynamic model of standing-up was developed. In each particular subject, the optimization criterion which yielded trajectories that best resemble the measured standing-up movement was determined. Since the intact persons used considerably different criteria in choosing the standing-up trajectories than the amputees, the optimal trajectories were computed by minimizing cost functionals (CF) with distinctive structures for each group of individuals. In intact persons, a unique cost functional was found which yielded realistic standing-up manoeuvres. In amputees, subject-specific sets of parameters indicating slightly different preferences in optimizing the effort of particular muscle groups were used. PMID- 15536030 TI - Kinetics of compensatory gait in persons with myelomeningocele. AB - This study investigated the kinetic strategy and compensatory mechanisms during self-ambulatory gait in children with lumbo-sacral myelomeningocele. Thirty-one children with mid-lumbar to low-sacral myelomeningocele who walked without aids and 21 control children were evaluated by three-dimensional gait analysis. Joint moments in all planes at the hip and knee and sagittal moments at the ankle, as well as joint power and work done at all three joints, were analyzed. Joint moment capacity lost due to plantarflexor and dorsiflexor weakness was provided instead by orthotic support, but other joints were loaded more to compensate for the weakness at the ankles and restricted ankle motion. Subjects with total plantarflexor and dorsiflexor paresis and strength in the hip abductors had more knee extensor loading due to plantarflexor weakness and dorsiflexion angle of the orthotic ankle joint. The subjects with orthoses also generated more power at the hip to supplement the power generation lost to plantarflexor weakness and fixed ankles. The most determinant muscle whose paresis changes gait kinetics was the hip abductor. Hip abductor weakness resulted in a characteristic pattern where the hips displayed an eccentric adduction moment, mediating energy transfer into the lower limbs, and the hips replaced the knees as power absorbers in early stance. Joint moment, power and work analyses complement a kinematic analysis to provide a complete picture of how children who have muscle paresis recruit stronger muscle groups to compensate for weaker ones. PMID- 15536031 TI - Ground reaction forces on stairs: effects of stair inclination and age. AB - The goals of the study were to compare data of vertical ground reaction force (GRF) parameters during level walking, stair ascent and descent on three different stair inclinations and three different age groups. Twenty healthy subjects of three age groups (young 33.7 years; middle 63.6 years; old 76.5 years) were tested during the seven test conditions with 8-10 repetitions. Vertical forces were measured during two consecutive steps with force plates embedded in the walkway and the staircase. The results showed that during level walking the vertical GRF curves were very regular and repetitive, the trail-to trial variability and left-right asymmetry of defined test parameters being around 2-5% and 3-5%. During stair ascent the vertical GRF force pattern was found to change slightly compared to level gait, but considerably compared to stair descent. On the steep stair the average vertical load increased up to 1.6 BW, and variability (5-10%) and asymmetry (5-15%) were increased significantly. The steep stair descent condition was found to be the most demanding test showing the largest variability and asymmetry and thus, the least stable gait pattern. Age was found to be a factor which should be considered, because the young age group walked faster and produced larger vertical GRF maxima during level walking and on stair ascent than the middle and old age group. Differences between the middle and old age group were found to be small. The present investigation is the first to provide normative data of GRF parameters on gait variability and symmetry of two consecutive steps during level gait and stair ambulation. It is the intention that the results of this study may be used as a basis for comparison with patient data. PMID- 15536032 TI - Direct visualisation of gaze and hypometric saccades in cerebellar patients during visually guided stepping. AB - Four patients suffering from primary cerebellar degeneration and healthy matched controls undertook a test of functional mobility that demanded precise foot placement at each step. Vertical and horizontal eye movements were measured (using a head mounted eye tracking system) together with footfall patterns. Healthy subjects stepped accurately onto all targets and produced a clear pattern of saccadic eye movements, fixating each target in the sequence just prior to footlift. Still video frames, showing direction of gaze while walking, provide direct visual confirmation that these saccades serve to transfer gaze between successive targets in the walkway sequence. The planning of the saccade to the next target probably provides the locomotor control system with information useful for planning the corresponding (and shortly following) step. Cerebellar patients showed characteristic locomotor and oculomotor deficits. Dysmetric saccades to fixate footfall targets were seen in 39% of steps. Analysis confirms that these multi-saccadic eye movements include an initial hypometric saccade, which undershoots the target, followed by one or more additional saccades in the same direction. Direct visualisation of gaze at the end of a saccadic sequence confirms that these additional saccades are indeed corrective resulting in a foveal image of the footfall target. PMID- 15536033 TI - The assessment of body sway and the choice of the stability parameter(s). AB - This methodological study aims at comparison of the practical usefulness of several parameters of body sway derived from recordings of the center of pressure (CoP) with the aid of a static force platform as proposed in the literature. These included: mean displacement velocity, maximal range of movement along x- and y-co-ordinates, movement area, planar deviation, phase plane parameter of Riley and the parameters of the diffusion stabilogram according to Collins. They were compared in over 850 experiments in a group of young healthy subjects (n = 10, age 21-45 years), a group of elderly healthy (n = 38, age 61-78 years) and two groups of elderly subjects (n = 10 and n = 21, age 65-89 years) with stability problems under different conditions known to interfere with stability as compared to standing with open eyes fixing a visual anchoring point: closing the eyes, standing on plastic foam in stead of a firm surface and performing a cognitive task: the modified stroop test. A force platform (Kistler) was used and co-ordinates of the body's center of pressure were recorded during 60 s of quiet barefoot standing with a sampling frequency of 10 Hz. In general, the results show important overlapping among groups and test conditions. Mean displacement velocity shows the most consistent differences between test situations, health conditions and age ranges, but is not affected by an extra cognitive task in healthy old people. Mean maximal sideways sway range is different among groups and test conditions except for the cognitive task in young and elderly subjects. Standardised displacement parameters such as standard deviations of displacements and planar deviation discriminate less well than the actual range of motion or the velocity. The critical time interval derived from the diffusion stabilogram according to Collins et al. seems to add a specific type of information since it shows significant influence from addition of a cognitive task in old subjects standing on a firm surface but not when standing on plastic foam. The critical time interval shows no consistent relation to any other parameter. The influence of cognitive activity on balance merits further exploration. A new parameter, sum of maximal deviation time (SDT) was proposed showing complete discrimination between frail elderly and other old subjects when obtained while visual input was suppressed. It was concluded that mean displacement velocity seems to be the most informative parameter in most situations. PMID- 15536034 TI - Does head extension and flexion increase postural instability in elderly subjects when visual information is kept constant? AB - The present study determined the effects of flexing and extending the head on the postural stability and mean anterior-posterior (A-P) center of mass (CM) position during upright stance in the elderly. To ensure visual input to stability was not a confounding variable, visual information was kept as constant as possible for all head positions. Twelve healthy elderly subjects (72.3 +/- 4.7 years) were asked to stand stationary on a single force-platform. Postural stability (assessed using the rms A-P excursion of the center of pressure (CP)) was determined for standing with the head erect, and with the head flexed and extended. The vestibular contribution to postural stability becomes increasingly important under challenging conditions, so to highlight the effects of vestibular system input, measurements of postural stability under conditions where visual and somatosensory inputs were disrupted were included. Changes in the mean A-P CM position when tilting the head were assessed by determining changes in the mean A P location of the CP from standing with the head erect. Compared to standing with the head erect and looking straight ahead, postural stability was reduced when the head was flexed or extended (P < 0.01). Changes in mean A-P CM position were only significant when standing with the head flexed (P < 0.05). This suggests that increases in postural instability with the head tilted from the erect position may be in part due to mechanical perturbation rather than solely vestibular disruption. PMID- 15536035 TI - The effects of blurred vision on the mechanics of landing during stepping down by the elderly. AB - Visual impairment is an important risk factor for falls. However, relatively little is known about how visual impairment affects stair or step negotiation. The aim of the present study was to determine the effects of blurred vision on the mechanics of landing during stepping down by the elderly. Twelve elderly subjects (72.3 +/- 4.7 year) stepped down from three levels (7.2 cm, 14.4 cm and 21.6 cm). Step execution time, ankle and knee joint angular displacements at the instance of ground contact, and vertical landing stiffness and the amount of bodyweight supported by the contralateral (support) limb during the initial contact period were recorded. Measurements were repeated with vision blurred by light scattering lenses. With blurred vision, step execution time increased (P < 0.05), knee flexion and ankle plantar-flexion increased (P < 0.05), vertical stiffness decreased (P < 0.01), and the amount of bodyweight being supported by the contralateral leg increased (P < 0.05). These findings suggest that under conditions of blurred vision, subjects were more cautious and attempted to 'feel' their way to the floor rather than 'drop' on to it. This may have been an adaptation to increase the kinaesthetic information from the lower limb to make up for the unreliable or incomplete visual information. Correcting common visual problems such as uncorrected refractive errors and cataract may be an important intervention strategy in improving how the elderly negotiate stairs. PMID- 15536036 TI - Familiarisation to treadmill walking in unimpaired older people. AB - We studied the amount of time required for treadmill familiarisation in older people and also whether familiarised treadmill walking could be generalised to overground walking. Sixteen healthy volunteers over 65 years of age walked on a level overground walkway and on a treadmill at the same speed for up to 15 min. A motion measurement system was used to measure the sagittal-plane kinematics of the knee and cadence during overground walking and after 0, 2, 4, 6, 8, 10, 12 and 14 min of treadmill walking. Older adults had not familiarised to the treadmill within 15 min as many participants continued to hold the treadmill's handrails and as reliability and absolute difference scores were still changing. Participants were most familiarised after 14 min on the treadmill. Furthermore, treadmill walking after 14 min was not closely related to overground walking in older adults, with measures on the treadmill only being able to predict knee angles during overground to within 8.0 degrees , or cadence to within 16.6 steps/min with 95% confidence. Treadmill walking in older adults after a single 15-min training session could not be generalised to overground walking. PMID- 15536037 TI - Within- and between-day stability of treadmill walking VO2 in children with hemiplegic cerebral palsy, stability of walking VO2 in children with CP. AB - Within- and between-day stability in locomotor energy use was quantified in 13 children with hemiplegic cerebral palsy (CP). During testing, subjects were familiarized with the laboratory environment (Session 1), performed three 5 min level treadmill walks (Trials 1-3) at 0.67 m*s(-1) (Session 2), and completed a single 5 min walk (Trial 4) at 0.67 m*s(-1) (Session 3). In Sessions 2 and 3, heart rate (HR) was assessed and expired air was collected and analyzed to determine VO2. Data analyses revealed no significant difference (P > 0.05) in either net VO2 (ml kg(-1)*min(-1)) or EEI(HR) (b*m(-1)) across the three trials performed in Session 2 and between average measures of net VO2 and EEI(HR) quantified in Session 2 and those obtained in Session 3. Mean within-day coefficient of variation (CV) values for net VO2 and EEI(HR) were 8.6% +/- 8.5% and 13.9% +/- 7.8%, respectively. Analysis of between-day variability and energy expenditure revealed a between-day CV value of 13.1% for net VO2 and 24.0% for EEI(HR). In addition, significant inverse relationships between Gross Motor Function Measure (GMFM) scores and within- (r = -0.61) and between-day (r = 0.58) CV values for net VO2 were detected. Viewed in concert, these data suggest that fairly stable within- and between-day measures of locomotor energy expenditure during level treadmill walking can be achieved in subjects with hemiplegic CP if testing is preceded by a short period of treadmill accommodation. However, children with greater motor dysfunction may require an extended period of treadmill accommodation to reduce trial-to-trial variability in walking energy use. PMID- 15536038 TI - Review of motor control mechanisms underlying impact absorption from falls. AB - The absorption of impacts resulting from contact with a landing surface during gait, running and drop landings has received considerable attention in the literature. This research has important clinical relevance as failure to appropriately plan and control impact absorption may lead to injuries to the musculo-skeletal system. This review attempts to summarize evidence gathered by studies on the motor control aspects of impact absorption during landing movements. Although this review focuses primarily on the control of landings from self-initiated falls or 'drop landings', an understanding of the motor control mechanisms underlying impact absorption is essential to understand common anticipatory and reflex mechanisms involved in a broader variety of movements such as running and jumping. The review is structured in three parts: the first two parts examine the preparatory muscle activity occurring during the fall (Part I) and after touch down (Part II). Part III explores the proposed sensorimotor mechanisms underlying the control of landing. The review concludes with as yet unresolved questions and directions for future research. PMID- 15536039 TI - The natural history of human gait and posture. Part 1. Spine and pelvis. AB - The human fossil record is one of the most complete for any mammal. A basal ancestral species, Australopithecus afarensis, exhibits a well-preserved postcranium that permits reconstruction of important events in the evolution of our locomotor skeleton. When compared with those of living apes and modern humans, this species provides a number of insights into the origin and design of the modern human frame as well as the selective agencies that have guided its evolution during the past three million years. Evolutionary aspects of the human spine and pelvis are reviewed, including their impact on several clinically relevant aspects of human gait and posture. PMID- 15536040 TI - The natural history of human gait and posture. Part 2. Hip and thigh. AB - The human fossil record is one of the most complete for any mammal. A basal ancestral species, Australopithecus afarensis, exhibits a well-preserved postcranium that permits reconstruction of important events in the evolution of our locomotor skeleton. When compared to those of living apes and humans, it provides insights into the origin and design of the modern human frame. Evolutionary aspects of the human hip and thigh are reviewed, including the unusual corticotrabecular structure of the human proximal femur, and our markedly elongated lower limb. It is postulated that the latter may be more related to birthing capacity than to locomotion. PMID- 15536041 TI - Gait analysis system for assessment of dynamic loading axis of the knee. AB - The purpose of this study was (1) to demonstrate a computer-assisted gait analysis system that can visualize the locus of the dynamic loading axis on the proximal tibia joint surface, and (2) to assess the accuracy of this system in a patient with bilateral knee osteoarthritis (OA). This system uses force plate data, CT skeletal structure data and motion capture data obtained from an infrared position sensor. The relative positions between bones and markers were used to calculate skeletal model movement based on movement of the markers. The locus of the dynamic loading axis on the knee joint was defined as the point on the proximal tibia joint surface that intersected with the loading axis of the lower limb, which passed through the centre of the femoral head and the centroid of multiple points surrounded by the distal tibia joint surface contour. To assess the accuracy of this system, open MRI was used to evaluate positions of skin markers against bones in six healthy volunteers. The locus in a patient was affected by differences between the varus knee with medial compartment OA on the non-operative side and the knee treated with high tibial osteotomy (HTO) on the opposite side. At knee flexion angles of 0 degrees, 15 degrees and 30 degrees, the mean value of measurement error for point locations on the locus was within 5.6% of joint width in the lateral direction (JWLD) on the proximal tibia joint. This system can provide clinically useful information for evaluation of the dynamic loading axis on the knee joint surface. PMID- 15536042 TI - Assessment of the viral safety of antivenoms fractionated from equine plasma. AB - Antivenoms are preparations of intact or fragmented (F(ab')2 or Fab) immunoglobulin G (IgG) used in human medicine to treat the severe envenomings resulting from the bites and stings of various animals, such as snakes, spiders, scorpions, or marine animals, or from the contact with poisonous plants. They are obtained by fractionating plasma collected from immunized horses or, less frequently, sheep. Manufacturing processes usually include pepsin digestion at acid pH, papain digestion, ammonium sulphate precipitation, caprylic acid precipitation, heat coagulation and/or chromatography. Most production processes do not have deliberately introduced viral inactivation or removal treatments, but antivenoms have never been found to transmit viruses to humans. Nevertheless, the recent examples of zoonotic diseases highlight the need to perform a careful assessment of the viral safety of antivenoms. This paper reviews the characteristics of equine viruses of antivenoms and discusses the potential of some manufacturing steps to avoid risks of viral contamination. Analysis of production parameters indicate that acid pH treatments and caprylic acid precipitations, which have been validated for the manufacture of some human IgG products, appear to provide the best potential for viral inactivation of antivenoms. As many manufacturers of antivenoms located in developing countries lack the resources to conduct formal viral validation studies, it is hoped that this review will help in the scientific understanding of the viral safety factors of antivenoms, in the controlled implementation of the manufacturing steps with expected impact on viral safety, and in the overall reinforcement of good manufacturing practices of these essential therapeutic products. PMID- 15536043 TI - Potency of pertussis component in the DTP vaccine--an overview of three decade study in Poland. AB - In Poland, similar to many highly immunized Western countries, a recent increase in cases of pertussis has been observed. This study aims to evaluate the level of potency fluctuations of the pertussis component of Polish-produced DTP vaccine due to the changes having occurred in production and potency testing procedures. We compared the potency of the pertussis component of DTP vaccine lots produced and evaluated in similar periods and with similar production and testing procedures. Records of Kendrick test results performed over a 30-year period were available for analysis. This study confirms the role of different manufacturers, changes in vaccine strain compositions, in-house reference preparations used as reference vaccines in the Kendrick tests, and in mice of single strain sources in the potency values obtained. In addition, the comparisons performed revealed a downtrend in potency levels since 1992. Potency decrease in vaccine lots produced during 1992-1997 has been positively correlated to the lowering of the number of IOU/dose. Strain compositions of the DTP vaccine pertussis component and in-house references have been found to be associated with the fluctuation in potency estimations, and confirmed their crucial role in ensuring vaccine efficacy. Our study reveals that relative efficacy of the DTP vaccine produced in 1992-1997 might be lower than that of vaccines produced in other periods. This might in turn explain the increase in pertussis cases among children aged 5-15 years which is presently being observed in Poland. PMID- 15536044 TI - An improved method for detection of replication-competent retrovirus in retrovirus vector products. AB - Contamination by replication-competent retrovirus (RCR) is one of the most important safety issues of retrovirus vector products for gene therapy clinical research. To improve the sensitivity of RCR detection and to shorten the assay period, we have developed a novel RCR detection method (infectivity RT-PCR method) based on real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) in combination with virus infection and a novel virus concentration method using polyethyleneimine (PEI)-conjugated magnetic beads. In this method, permissive cells were infected with RCR samples, and amplified RCR in the culture supernatants was adsorbed by PEI-beads. Then RCR RNA extracted from PEI-beads was quantified by real-time RT-PCR. We demonstrated that 1 infectious unit (iu) of RCR spiked in 10(6) cfu/ml of vector products could be detected within 3 days, and the sensitivity for viral detection was increased 3- to 10-fold compared with the direct S+L- assay. By this method, the presence of retroviral vector interfered with RCR detection only slightly. In conclusion, infectivity RT-PCR conducted in conjunction with virus concentration using PEI beads can detect RCR more sensitively and rapidly than the conventional infectivity assay. PMID- 15536045 TI - Neurovirulence of mumps virus: intraspinal inoculation test in marmosets. AB - An intraspinal inoculation test of mumps virus using marmosets was performed in order to develop a neurovirulence test of mumps vaccines. In the group inoculated with non-neurovirulent Jeryl Lynn vaccine strain at 10(2.0) pfu/dose, there were only minimal histopathological changes in 3 of the 5 marmosets. In contrast, all marmosets inoculated with neurovirulent Urabe and NK-M46 vaccine strains developed extensive encephalitis and meningitis. Thus, this marmoset model, which can distinguish between non-neurovirulent and neurovirulent vaccine strains, is useful for evaluating neurovirulence of vaccine strains and elucidating the molecular pathogenesis of mumps. PMID- 15536046 TI - Validation of binary ethyleneimine (BEI) used as an inactivant for foot and mouth disease tissue culture vaccine. AB - The complete inactivation of Foot and Mouth Disease (FMD) virus is a critical requirement in the production of FMD vaccine to ensure the safety of the product. Binary ethyleneimine (BEI) is an aziridine compound, produced from bromoethylamine hydrobromide (BEA) commonly used for the inactivation of FMD virus during vaccine manufacturing. The validation of BEI, when used as an inactivant, is essential to ensure the quality of the inactivating agent and the validity of the process. In the present study, the inactivation kinetics of Foot and Mouth Disease virus (O, A and Asia-1 serotypes) were determined for different concentrations of BEI (0.4 mM, 0.8 mM, 1.2 mM, 1.6 mM and 2.0 mM). Statistically significant differences in the inactivation kinetics were observed between 0.4 mM and 1.6 mM of BEI. The results indicated that BEI at 1.6 mM was able to inactivate the FMD virus within 8-10 h. Increasing the concentration of BEI beyond 1.6 mM did not appreciably improve the inactivation process. No differences in the inactivation kinetics were found between the various serotypes studied. This study can be used as a guideline for routine procedures for validating the quality of BEA and the inactivation process. PMID- 15536047 TI - Validation of an anti-PA-ELISA for the potency testing of anthrax vaccine in mice. AB - The potency test for the anthrax vaccine currently licensed for human use in the United States (Anthrax Vaccine Adsorbed) involves the protection of actively immunized guinea pigs from a lethal challenge with a virulent strain of Bacillus anthracis. Lethal challenge tests entail the use of specialized containment facilities for the safe and secure handling of the challenge strain. This potential difficulty, plus humane considerations, have prompted us to investigate non-lethal, alternative immunogenicity assays that could be considered as potency tests not only for the current vaccine, but also for vaccines under development. Immunogenicity tests will require suitable measurement of an antibody response to relevant antigens, by methods such as enzyme linked immunosorbent assay (ELISA) or a toxin neutralization assay. Any assay chosen for this purpose should be adequately validated and reproducible by other laboratories. Validation of an analytical procedure requires the demonstration that the assay is suitable for its intended purpose. The objective of this work was to study the performance of an anti-PA-ELISA designed to assess the antibody response to anthrax vaccines in mice. Validation studies were performed according to the guidelines of the International Conference of Harmonization (ICH), and we have established the working range of the assay (37-1159 EU/mL) on the bases of the following parameters: linearity (20-1159 EU/mL; r2=0.99; p-value=0.21), accuracy (91-118% recovery), precision (< or =20%CV, repeatability; < or =9 and < or =21%CV, intermediate precision per day and per analyst, respectively), detection limit (5 EU/mL), and quantification limit (37 EU/mL). We believe that assay specificity and the above characteristics are adequate to allow this ELISA to be considered for use in a mouse immunogenicity (potency) test of anthrax vaccines, and for the standardization of reagents. PMID- 15536048 TI - Impact of manufacturing, irradiation and filtration steps to bacterial contamination of autologous fibrin sealant. AB - BACKGROUND: Preoperative production of autologous fibrin sealant has become a routine procedure during the last years. As a certain percentage of blood products is contaminated with bacteria, contamination of plasma used for the production of fibrin sealant cannot be excluded. Especially in the orthopaedic setting, application of contaminated fibrin sealant can cause severe infections. MATERIALS AND METHODS: We contaminated plasma with Staphylococcus epidermidis, Corynebacterium striatum, Bacillus subtilis or Escherichia coli and produced fibrin sealant by cryoprecipitation and alcohol precipitation. Additionally, the products were gamma-irradiated at a dose of 30 Gy, frozen at -55 degrees C and filtered through a 0.2 microm filter after thawing. After each preparation step, samples were drawn and numbers of colony forming units were counted after incubation on agar plates. RESULTS: Cryoprecipitation, irradiation, freezing at 55 degrees C, and alcohol precipitation have only little impact on numbers of colony forming units. Filtration through a bacterial filter results in a sterile product. CONCLUSION: Bacteria in plasma as a starting material for production of fibrin sealant survive all routine steps of production, including gamma irradiation and freezing. Filtration of the product through a qualified bacterial filter is the only safe means to provide a sterile product. PMID- 15536049 TI - Seasonal variations of a battery of biomarkers and physiological indices for the mussel Mytilus galloprovincialis transplanted into the northwest Mediterranean Sea. AB - Seasonal variations of six mussel (Mytilus galloprovincialis) biomarkers at two sites in the Mediterranean Sea were compared with physiological indices (condition, growth and gonad maturation), environmental parameters (temperature, salinity and turbidity), and chemical contamination levels. The basal levels of acetylcholinesterase (AChE), DNA adducts, benzo[a]pyrene hydroxylase (BPH), heat shock proteins (HSP70), metallothioneins (MT) and P-glycoprotein (P-gp)-mediated multixenobiotic resistance (MXR) were estimated as early warning signals in caged mussels sampled at Carteau (native site) and La Fourcade (transplantation site) over a 2-year period. The Carteau and La Fourcade mussels have specific chemical contamination profiles but a similar range of values. For example, both are highly contaminated by heavy metals (201 and 258.4 mg kg(-1) dw, respectively) and considered as moderately impacted for polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). However, contamination levels at Carteau are twice as high for PAHs (101.5 mg g(-1) dw) and PCBs (90.2 mg g(-1) dw) than La Fourcade. The seasonal contamination trend at Carteau showed six-fold higher levels of pyrolytic pollutants in winter. Although few tissue lesions were detected in individuals studied at either site, greater parasitic infestation was observed at Carteau. The results of findings from the two Mediterranean pilot studies support the adaptability of transplanted mussels to be used as biomarkers and to establish physiological endpoints for chemical contaminant exposure. PMID- 15536050 TI - Effect of crotapotin on the biological activity of Asp49 and Lys49 phospholipases A(2) from Bothrops snake venoms. AB - Myonecrosis, in addition to edema and other biological manifestations, are conspicuous effects of Bothrops snake venoms, some of them caused by phospholipases A(2) (PLA(2)s). Asp49-PLA(2)s are catalytically active, whereas Lys49-PLA(2)s, although highly toxic, have little or no enzymatic activity upon artificial substrates, due to a substitution of lysine for aspartic acid at position 49. Crotapotin (CA), the acidic counterpart of crotoxin PLA(2) (CB), is a PLA(2)-like protein from Crotalus durissus terrificus snake venom, and is considered a chaperone protein for CB, able to increase its lethality about ten fold, but to inhibit the formation of the rat paw edema induced by carrageenin and by snake venoms. In this study, we demonstrate that CA significantly inhibits the edema induced by BthTX-I (23% inhibition), BthTX-II (27%), PrTX-I (25%), PrTX III (35%) and MjTX-II (10%) on the mouse paw. CK levels evoked by isolated Asp49 or Lys49-PLA(2)s were reduced by 40% to 54% in the presence of CA and, in all cases, the membrane damaging activity of the toxins was also reduced. Circular dichroism spectra of the PLA(2)s in the presence and absence of CA showed that there was not any detectable secondary structural modification due to association between CA and the myotoxins. However, Fourier Transformed Infrared (FT-IR) analysis indicated that ionic and hydrophobic contacts contributed to stabilize this interaction. PMID- 15536051 TI - Macrocyclic lactones: distribution in plasma lipoproteins of several animal species including humans. AB - We studied the in vitro distribution of macrocyclic lactones (MLs), lipophilic anthelmintic drugs, in the plasma of several animal species including humans. First, in vitro spiking of goat plasma was performed with ivermectin, moxidectin, abamectin, doramectin, or eprinomectin. In parallel, goats were treated with subcutaneous injection of ivermectin. Then, cow, sheep, rabbit, pig, and human plasma were spiked with moxidectin. Four fractions were separated using KBr density gradient ultracentrifugation: very-low-density lipoprotein (VLDL), low density lipoprotein (LDL), high-density lipoprotein (HDL), and lipoprotein deficient fraction. Cholesterol was analyzed by enzymatic assay and MLs by high performance liquid chromatography. An average of 96% of MLs was associated with lipoproteins. The five MLs studied distributed similarly into goat plasma fractions with a preferential association with HDL (80-90%). Ivermectin partitioning in goat plasma was similar after in vitro spiking and in vivo treatment. In species displaying various lipoprotein profiles, moxidectin was also mainly associated with HDL. However, in human plasma, moxidectin was associated with a lesser extent to HDL (70%) and more to LDL (22%) when compared to other animal species. A relation between the plasma cholesterol content and pharmacokinetics of the drug is suggested. Our finding will allow further exploration of intestinal lymphatic absorption and milk elimination of these compounds-mechanisms in which lipoproteins are involved. In addition, possible improvements of new drug delivery systems are suggested. PMID- 15536052 TI - Influence of beta-carotene on lysosomal hydrolases and their natural substrates in major salivary glands of hamsters treated with 7,12-dimethylbenzanthracene (DMBA). AB - We evaluated the effects of beta-carotene, a precursor of vitamin A, on the activity of some lysosomal hydrolases and on the levels of their natural substrates in hamster major salivary glands during experimental oral 7,12 dimethylbenzanthracene (DMBA) carcinogenesis. Sixty-four hamsters (Cricetus auratus) were divided into four groups--group 1: untreated control; group 2: DMBA was painted three times a week in the left buccal pouch; group 3: beta-carotene was painted three times a week in the left buccal pouch; group 4: DMBA and beta carotene were painted alternatively in the left buccal pouch. After 16 weeks, the animals were sacrificed and the activities of some lysosomal hydrolases and their natural substrates in the major salivary glands were measured. beta-Carotene when administered topically in DMBA treated animals (group 4) reduced the levels of the majority of enzymes and substrates closer to those of the untreated control group, thus outlining a mild protective effect of beta-carotene towards the DMBA carcinogenic stress. Nevertheless, the presence of some enzymes which responded negatively to the combined administration of DMBA and beta-carotene suggests the necessity for future studies on the effect of beta-carotene at different concentrations, the systemic administration and the possibility to combine the topical beta-carotene administration with other chemopreventive drugs. PMID- 15536053 TI - Evaluation of PAH bioaccumulation and DNA damage in mussels (Mytilus galloprovincialis) exposed to spilled Prestige crude oil. AB - We analyzed the hydrocarbon composition of the Prestige oil as it reached the shores, its solubility in sea water, its bioaccumulation, and the genotoxic damage associated to oil exposure, using Mytilus galloprovincialis as sentinel organism. Mussels were exposed to two oil volumetric ratios (1:500 and 2:500) for 12 days. Great concentrations of total polycyclic aromatic hydrocarbons (TPAH) have been obtained, being in general higher in the samples from the dose of 1:500, both in sea water (55.14 vs. 41.96 microg/l) and mussel tissue (16,993.80 vs. 17,033.00 microg/kg), probably due to the great tendency of these compounds to link to particles in water. Comet assay results reflected an increase in the DNA damage associated to oil exposure, higher in the mussels exposed to the higher aqueous TPAH content. In the view of our results, the importance of the evaluation of biodisponibility, bioaccumulation and DNA damage in the assessment of the effects of xenobiotic pollutants to marine environments could be highlighted. PMID- 15536054 TI - 2,3,7,8-Tetrachlorodibenzo-p-dioxin elicits aryl hydrocarbon receptor-mediated apoptosis in the avian DT40 pre-B-cell line through activation of caspases 9 and 3. AB - The halogenated aromatic hydrocarbon 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is known to induce immunotoxicity, but relatively little is known regarding its effects on B-lymphocytes, and on avian B-cells in particular. In this study, the avian bursal pre-B-cell line DT40 was exposed to TCDD ranging from 1 to 500 nM for 1 and 6 h. At 100 nM, TCDD caused a significant increase in the number of apoptotic cells, as assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) assay, and induced the expression of the chicken cytochrome P450 1A4 (CYP1A4) mRNA, a hallmark of TCDD exposure. TCDD induced transient upregulation of aryl hydrocarbon receptor (AhR) mRNA. At 100 nM, both caspase 3 and caspase 9 were transiently upregulated after 1 h, but returned to normal levels after 6 h of exposure. Challenge with TCDD after AhR blockade with resveratrol, a competitive AhR antagonist, prevented changes in caspases 3 and 9 and in the AhR message itself, suggesting that the effects of TCDD were mediated via the AhR. TCDD did not cause significant changes in the relative gene expression of caspase 8, Bcl-2 and Bcl-xL. We conclude that avian DT40 pre-B cells exposed to TCDD are susceptible to apoptosis, likely through activation of executioner caspase 3. PMID- 15536055 TI - The serotonergic system is involved in feeding inhibition by pymetrozine. Comparative studies on a locust (Locusta migratoria) and an aphid (Myzus persicae). AB - Pymetrozine inhibits feeding in aphids immediately after application without producing visible neurotoxic effects, as previously reported. In the present work, Locusta migratoria, though not a plant-sucking insect, was found to respond to pymetrozine by displaying unique symptoms, which were lifting and stretching of the hindlegs, in addition to inhibition of feeding. In locust, pymetrozine enhanced spontaneous spike discharge of the metathoracic and suboesophageal ganglia in situ at nanomolar concentrations. Similarly, pymetrozine increased the spontaneous rhythmic contractions of the isolated foregut with maximal effects also in the nanomolar range. The actions of pymetrozine were counteracted by biogenic amine receptor antagonists and mimicked by serotonin, not by dopamine and octopamine. Moreover, pymetrozine and serotonin strongly potentiated the effects of each other. Pymetrozine was inactive at all neurotransmitter receptors present on isolated locust neuronal somata, and at all other examined neuronal sites. In Myzus persicae, electrical penetration graph experiments revealed that serotonin, like pymetrozine, inhibited stylet penetration, and strongly enhanced the action of pymetrozine, comparable to the locust. Amine receptor antagonists were not specifically active in the aphid. We conclude from the present results that pymetrozine acts via a novel mechanism that is linked to the signalling pathway of serotonin. PMID- 15536056 TI - Feeding behavior and venom toxicity of coral snake Micrurus nigrocinctus (Serpentes: Elapidae) on its natural prey in captivity. AB - The feeding behavior and venom toxicity of the coral snake Micrurus nigrocinctus (Serpentes: Elapidae) on its natural prey in captivity were investigated. Coral snakes searched for their prey (the colubrid snake Geophis godmani) in the cages. Once their preys were located, coral snakes stroke them with a rapid forward movement, biting predominantly in the anterior region of the body. In order to assess the role of venom in prey restraint and ingestion, a group of coral snakes was 'milked' in order to drastically reduce the venom content in their glands. Significant differences were observed between snakes with venom, i.e., 'nonmilked' snakes, and 'milked' snakes regarding their behavior after the bite. The former remained hold to the prey until paralysis was achieved, whereas the latter, in the absence of paralysis, moved their head towards the head of the prey and bit the skull to achieve prey immobilization by mechanical means. There were no significant differences in the time of ingestion between these two groups of coral snakes. Susceptibility to the lethal effect of coral snake venom greatly differed in four colubrid species; G. godmani showed the highest susceptibility, followed by Geophis brachycephalus, whereas Ninia psephota and Ninia maculata were highly resistant to this venom. In addition, the blood serum of N. maculata, but not that of G. brachycephalus, prolonged the time of death of mice injected with 2 LD(50)s of M. nigrocinctus venom, when venom and blood serum were incubated before testing. Subcutaneous injection of coral snake venom in G. godmani induced neurotoxicity and myotoxicity, without causing hemorrhage and without affecting heart and lungs. It is concluded that (a) M. nigrocinctus venom plays a role in prey immobilization, (b) venom induces neurotoxic and myotoxic effects in colubrid snakes which comprise part of their natural prey, and (c) some colubrid snakes of the genus Ninia present a conspicuous resistance to the toxic action of M. nigrocinctus venom. PMID- 15536057 TI - Differential responses of stress proteins, antioxidant enzymes, and photosynthetic efficiency to physiological stresses in the Florida red tide dinoflagellate, Karenia brevis. AB - This study identifies stress proteins and antioxidant enzymes that may play a role in the survival strategies of the Florida red tide dinoflagellate, Karenia brevis. Heat shock protein 60 (Hsp 60), mitochondrial small heat shock protein (mitosHsp), chloroplastic small heat shock protein (chlsHsp), Mn superoxide dismutase (SOD), and Fe SOD were first identified by Western blotting. The induction of these proteins in laboratory cultures in response to elevated temperatures, hydrogen peroxide, lead, or elevated light intensities was next assessed. In parallel, F(V)/F(M), a measurement of photosynthetic efficiency and common proxy of cellular stress, was determined. Hsp 60, Fe SOD, and Mn SOD were induced following exposure to elevated temperatures, hydrogen peroxide, or lead. MitosHsp responded only to heat, whereas chlsHsp responded only to H(2)O(2) induced stress. The expression of stress proteins and antioxidant enzymes appears to be a more sensitive indicator of heat or chemically induced stresses than F(V)/F(M). However, F(V)/F(M) decreased significantly in response to elevated light intensities that did not induce the expression of stress proteins. These results identify for the first time stress proteins and antioxidant enzymes in K. brevis, provide evidence for differential sensitivity of cellular organelles to various sources of stress, and confirm the presence of conserved stress responses observed across phyla in a dinoflagellate. PMID- 15536058 TI - Arsenobetaine in Atlantic salmon (Salmo salar L.): influence of seawater adaptation. AB - Glycine betaine has been suggested to improve the maintenance of ionic and osmotic homeostasis during seawater adaptation in teleost fish. Arsenobetaine may also behave as an osmolyte, due to its structural similarity to glycine betaine. The influence of seawater adaptation on intestinal uptake and muscle accumulation of arsenobetaine in the teleost Atlantic salmon (Salmo salar L.) was investigated. Atlantic salmon (freshwater and seawater adapted) were given a single oral dose of arsenobetaine, which was absorbed over the intestine within 6 h after exposure. Seawater adapted Atlantic salmon had significantly higher levels of accumulated arsenobetaine in blood compared to the freshwater adapted salmon. However, seawater adaptation had no effect on the levels of accumulated arsenobetaine in muscle tissue. Similar retention of the administered dose was found in muscle tissue in both freshwater and seawater adapted salmon, with 49+/ 6% and 50+/-10% retention after 144 h, respectively. Results indicate that muscle retention was not influenced by salinity in seawater adapting teleosts. PMID- 15536059 TI - Shell protein characteristics and vitellogenin-like proteins in brine shrimp Artemia franciscana exposed to municipal effluent and 20-hydroxyecdysone. AB - We developed biomarkers to monitor the endocrine-disrupting potential of contaminants and municipal effluents in aquatic arthropods. Artemia fransciscana shrimp were cultured and exposed to increasing concentrations of 20 hydroxyecdysone (20HE) and solid phase (C-8) municipal effluent extract (MEE) for 48 h at 20 degrees C. The levels of vitellogenin (Vtg)-like proteins, alkali labile phosphates in total proteins and acetylcholinesterase (AChE) activity were determined in soft tissues. The levels of acid-soluble, alkali-soluble (sclerotin), neutral-soluble (arthropodin) proteins and chitin were determined to characterize the maturation state of shells. Both 20HE and the municipal effluent extract readily increased the total activity of acetylcholinesterase, alkali labile phosphates in proteins and vitellogenin-like proteins in brine shrimp. In shells, 20HE and the effluent extract increased the proportion of chitin in shells and acid-soluble proteins but were not statistically significant for the latter. The proportion of sclerotin was increased by 20HE but was not changed by the effluent extract. The proportion of arthropodin was decreased by both 20HE and the effluent extract. Correlation analysis revealed that, as expected, the proportion of acid-soluble protein for biomineralisation and chitin levels were positively correlated and the proportion of arthropodin and sclerotin were negatively correlated in shells exposed to 20HE. Principal component analysis revealed that the proportion of chitin, arthropodin, sclerotin and acid-soluble proteins explained 63% of the responses. Shell protein and carbohydrate contents, controlled by the molting hormone 20HE, could be used as efficient markers for ecdysial properties of various contaminants. The municipal effluent appears to contain ecdysial-like chemicals that are capable of producing changes in shell protein composition that are similar to 20HE. PMID- 15536060 TI - Free radical scavengers in susceptible/resistant Biomphalaria alexandrina snails before and after infection. AB - The activities of catalase (Cat), superoxide dismutase (SOD), glutathione peroxidase (GSHPx), glutathione transferase (GST), glucose-6-phosphate dehydrogenase (G6PD) and glyceraldehyde3-phosphate dehydrogenase (G3PD) were studied in tissue and hemolymph of susceptible (S) (EgBS(2)) and resistant (R) (EgBR(2)) Biomphalaria alexandrina snails. The results showed that CAT and GST were higher in the hemolymph of snails susceptible to Schistosoma mansoni than in that of snails resistant to infestation, while SOD and G3PD were lower in the susceptible snails. The role of these enzymes as free radical scavengers was traced 1 and 24 h after infection of the two snail lines with S. mansoni. Moreover, the activities of SOD and G3PD were also measured 2 and 4 weeks post infection. The results revealed that the overall enzymatic activities were higher in susceptible than in resistant snail tissues. After 1 h of infection, all enzymes were increased in R and S snails except GST and G6PD which decreased in S snails. After 24 h of infection, GST increased in S snails and G3PD decreased in both S and R snails while other enzymes reached normal levels. PMID- 15536061 TI - ELISAs for detecting vitellogenin in the fathead minnow (Pimephales promelas)-a critical analysis. Response to Mylchreest et al., Comp Biochem Physiol C 134: 251 257, 2003. PMID- 15536064 TI - Association between pain in the hip region and radiographic changes of osteoarthritis: results from a population-based study. AB - OBJECTIVES: The relationship between hip pain and radiographic change in the population is unclear due to lack of agreed definition for hip pain and difficulties in obtaining radiographs from asymptomatic random samples. Our objective was to assess the relationship between hip pain and radiographic change in osteoarthritis (OA) in a population sample aged over 45. METHODS: One thousand and seventy-one responders to a postal questionnaire using a recently validated approach to defining hip pain were stratified into hip pain-positive and negative groups and samples of each were X-rayed and scored for OA using both minimum joint space and the Croft score. The association between pain and X-ray score was estimated, weighting back to the age and gender distribution of the original population. RESULTS: Hip pain prevalence was 7% in males and 10% in females. Severe OA was present in 16% of those with and 3% of those without pain. Adjusting for age and gender, there was a very strong association of pain with severe OA [odds ratio (OR) 17.4, 95% confidence interval (CI) 3.0-102], but no association with mild/moderate OA (OR 1.4, 95% CI 0.4-4.7). By contrast, only 22% of men aged 45-54 with severe OA had current pain, though in older age groups the proportions with pain were higher (54-70%). CONCLUSIONS: Hip pain is relatively infrequent in the general population compared with the published reports of other regional pain syndromes. Mild/moderate radiographic change is very frequent and not related to pain, whereas severe change is rare but strongly related. In younger males, severe radiographic change is much less likely to be associated with pain. PMID- 15536063 TI - Finding new clock components: past and future. AB - The molecular mechanism of circadian clocks has been unraveled primarily by the use of phenotype-driven (forward) genetic analysis in a number of model systems. We are now in a position to consider what constitutes a clock component, whether we can establish criteria for clock components, and whether we have found most of the primary clock components. This perspective discusses clock genes and how genetics, molecular biology, and biochemistry have been used to find clock genes in the past and how they will be used in the future. PMID- 15536065 TI - Maternal and sibling microchimerism in twins and triplets discordant for neonatal lupus syndrome-congenital heart block. AB - OBJECTIVE: Neonatal lupus syndrome-congenital heart block (NLS-CHB) is an acquired autoimmune disease in which maternal autoantibodies are necessary but not sufficient for disease. Maternal myocardial cells have been found in the hearts of patients with NLS-CHB, suggesting that maternal microchimerism may also play a role. In this study we asked whether levels of microchimerism in the blood are associated with NLS-CHB in discordant twins and triplets. METHODS: Human leucocyte antigen (HLA)-specific and Y-chromosome-specific real-time quantitative polymerase chain reaction (PCR) was used to quantitatively assay maternal and sibling microchimerism in peripheral blood. Because of HLA allele sharing in families, it was not always possible to distinguish between multiple sources of microchimerism. RESULTS: In one family, maternal and/or sibling microchimerism was detected in two triplets who had CHB, but not in the triplet with transient hepatitis. Levels ranged from 4 to 948 genome-equivalents of foreign deoxyribonucleic acid per million host genome-equivalents (gEq/million). Over the first year levels of sibling microchimerism decreased in the triplet with complete CHB and increased in the triplet who progressed from first- to second degree CHB. In a second family, maternal and/or sibling microchimerism was detected in the healthy twin (1223 gEq/million) but not in the twin with CHB. CONCLUSIONS: Maternal and/or sibling microchimerism was detectable in the blood of infant twins and triplets discordant for NLS. Microchimerism in the blood was not specific for NLS-CHB, although in one family levels correlated with disease. Thus, microchimerism in the blood and/or tissues may be involved in the pathogenesis or progression of NLS-CHB, but additional factors must also contribute. Further investigation is warranted. PMID- 15536066 TI - Role for protein kinase Ctheta (PKCtheta) in TCR/CD28-mediated signaling through the canonical but not the non-canonical pathway for NF-kappaB activation. AB - NF-kappaB is a family of essential transcription factors involved in both embryonic development and inflammatory responses of the immune system. NF-kappaB can be activated by two pathways, i.e. the canonical (NF-kappaB1) pathway, which acts through the catalytic components of the IkappaB kinase complex and leads to IkappaB phosphorylation, degradation, and subsequent NF-kappaB nuclear translocation, or the non-canonical (NF-kappaB2) pathway, which involves NF kappaB-induced kinase-dependent proteolytic processing of p100/p52 to yield translocation-competent p52-containing NF-kappaB complexes. We examined the relative roles of the NF-kappaB1 and NF-kappaB2 pathways in TCR/CD28 costimulation. We found that TCR/CD28 costimulation activates the canonical but not the non-canonical NF-kappaB pathway and that the serine/threonine kinase protein kinase C (PKC) is essential for TCR/CD28-mediated canonical NF-kappaB activation in T cells. Importantly, TCR/CD28 costimulation induces higher p52 protein levels in T cells, but this effect is secondary to enhanced de novo synthesis of p100, not to enhanced processing of extant p100; PKC deficiency impairs signal-dependent p52 accumulation because of defects in p100 production. Finally, we found that TCR/CD28 costimulation induces IkappaBalpha, IkappaBbeta, and IkappaBepsilon degradation, and PKC is required for IkappaBalpha and IkappaBepsilon but not IkappaBbeta degradation. PKC acts solely within the canonical pathway to activate NF-kappaB, and PKC deficiency impacts upon p100/p52 processing in a manner that is independent of NF-kappaB-induced kinase. PMID- 15536067 TI - Pro-angiogenic signaling by the endothelial presence of CEACAM1. AB - Here, we demonstrate the expression of carcinoembryonic antigen-related cell adhesion molecule-1 (CEACAM1) in angiogenic sprouts but not in large mother blood vessels within tumor tissue. Correspondingly, only human microvascular endothelial cells involved in in vitro tube formation exhibit CEACAM1. CEACAM1 overexpressing versus CEACAM1-silenced human microvascular endothelial cells were used in migration and tube formation assays. CEACAM1-overexpressing microvascular endothelial cells showed prolonged survival and increased tube formation when they were stimulated with vascular endothelial growth factor (VEGF), whereas CEACAM1 silencing via small interfering RNA blocks these effects. Gene array and LightCycler analyses show an up-regulation of angiogenic factors such as VEGF, VEGF receptor 2, angiopoietin-1, angiopoietin-2, tie-2, angiogenin, and interleukin-8 but a down-regulation of collagen XVIII/endostatin and Tie-1 in CEACAM1-overexpressing microvascular endothelial cells. Western blot analyses confirm these results for VEGF and endostatin at the protein level. These results suggest that constitutive expression of CEACAM1 in microvascular endothelial cells switches them to an angiogenic phenotype, whereas CEACAM1 silencing apparently abrogates the VEGF-induced morphogenetic effects during capillary formation. Thus, strategies targeting the endothelial up-regulation of CEACAM1 might be promising for antiangiogenic tumor therapy. PMID- 15536068 TI - Serine racemase modulates intracellular D-serine levels through an alpha,beta elimination activity. AB - Mammalian brain contains high levels of d-serine, an endogenous co-agonist of N methyl D-aspartate type of glutamate receptors. D-Serine is synthesized by serine racemase, a brain enriched enzyme converting L- to D-serine. Degradation of D serine is achieved by D-amino acid oxidase, but this enzyme is not present in forebrain areas that are highly enriched in D-serine. We now report that serine racemase catalyzes the degradation of cellular D-serine itself, through the alpha,beta-elimination of water. The enzyme also catalyzes water alpha,beta elimination with L-serine and L-threonine. alpha,beta-Elimination with these substrates is observed both in vitro and in vivo. To investigate further the role of alpha,beta-elimination in regulating cellular D-serine, we generated a serine racemase mutant displaying selective impairment of alpha,beta-elimination activity (Q155D). Levels of D-serine synthesized by the Q155D mutant are several fold higher than the wild-type both in vitro and in vivo. This suggests that the alpha,beta-elimination reaction limits the achievable D-serine concentration in vivo. Additional mutants in vicinal residues (H152S, P153S, and N154F) similarly altered the partition between the alpha,beta-elimination and racemization reactions. alpha,beta-Elimination also competes with the reverse serine racemase reaction in vivo. Although the formation of L- from D-serine is readily detected in Q155D mutant-expressing cells incubated with physiological D-serine concentrations, reversal with wild-type serine racemase-expressing cells required much higher D-serine concentration. We propose that alpha,beta-elimination provides a novel mechanism for regulating intracellular D-serine levels, especially in brain areas that do not possess D-amino acid oxidase activity. Extracellular D-serine is more stable toward alpha,beta-elimination, likely due to physical separation from serine racemase and its elimination activity. PMID- 15536069 TI - A novel repressive E2F6 complex containing the polycomb group protein, EPC1, that interacts with EZH2 in a proliferation-specific manner. AB - The transcriptional repressor E2F6 has been identified as a component of two distinct polycomb group protein (PcG)-containing complexes, suggesting a mechanism for the recruitment of repressive complexes to target sequences in DNA. Whereas one complex is involved in the repression of classic E2F target genes in G0, a role for E2F6 within the cell cycle has yet to be defined. We searched for novel E2F6-binding proteins using a yeast two-hybrid screen and identified the PcG protein, EPC1. We showed that, both in vitro and in vivo, E2F6, DP1, and EPC1 form a stable core complex with repressive activity. Furthermore, we identified the proliferation-specific PcG, EZH2, as an EPC1-interacting protein. Using affinity purification, we showed that E2F6, DP1, EPC1, EZH2, and Sin3B co-elute, suggesting the identification of a novel E2F6 complex that exists in vivo in both normal and transformed human cell lines. EZH2 is required for cellular proliferation and consistent with this, EZH2 elutes with the E2F6-EPC1 complex only in proliferating cells. Thus we have identified a novel E2F6-PcG complex (E2F6-EPC1) that interacts with EZH2 and may regulate genes required for cell cycle progression. PMID- 15536070 TI - Heparin-binding epidermal growth factor-like growth factor links hepatocyte priming with cell cycle progression during liver regeneration. AB - The mechanisms that regulate the transition between the initial priming phase and DNA replication in liver regeneration are poorly understood. To study this transition, we compared events occurring after standard two-thirds partial hepatectomy, which elicits full regeneration, with response to a reduced hepatectomy, one-third partial hepatectomy (1/3PH), which leads to little DNA replication. Although the initial response to partial hepatectomy at the priming phase appeared to be similar between the two procedures, cell cycle progression was significantly blunted in 1/3PH mice. Among the main defects observed in 1/3PH mice were an almost complete deficiency in retinoblastoma phosphorylation and the lack of increase in kinase activity associated with cyclin E. We report that, in two-thirds partial hepatectomy mice, the expression of heparin-binding epidermal growth factor-like growth factor (HB-EGF) preceded the start of DNA replication and was not detectable in 1/3PH animals. Injection of HB-EGF into 1/3PH mice resulted in a >15-fold increase in DNA replication. Moreover, we show that hepatocyte DNA replication was delayed in HB-EGF knock-out mice. In summary, we show that HB-EGF is a key factor for hepatocyte progression through G(1)/S transition during liver regeneration. PMID- 15536071 TI - On the competition for available zinc. AB - Extended x-ray absorption fine structure (EXAFS) spectroscopy was combined with thermodynamic and kinetic approaches to investigate zinc binding to a zinc finger (C2H2) and a tetrathiolate (C4) peptide. Both peptides represent structural zinc sites of proteins and rapidly bind a single zinc ion with picomolar dissociation constants. In competition with EDTA the transfer of peptide-bound zinc ions proved to be 6 orders of magnitude faster than predicted for a dissociation association mechanism thus requiring ligand exchange mechanisms via peptide-zinc EDTA complexes. EXAFS spectra of C2H2 showed the expected Cys2His2-ligand geometry when fully loaded with zinc. For a 2-fold excess of peptide, however, the existence of zinc-bridged peptide-peptide complexes with dominating sulfur coordination could be clearly shown. Whereas zinc binding kinetics of C2H2 appeared as a simple second order process, the suggested mechanism for C4 comprises a zinc-bridged Zn-(C4)2 species as well as a Zn-C4 species with less than 4 metal-bound thiolates, which is supported by EXAFS results. A rapid equilibrium of bound and unbound states of individual ligands might explain the kinetic instability of zinc-peptide complexes, which enables fast ligand exchange during the encounter of occupied and unoccupied acceptor sites. Depending on relative concentrations and stabilities, this results in a rapid transfer of zinc ions in the virtual absence of free zinc ions, as seen for the zinc transfer to EDTA, or in the formation of zinc-bridged complexes, as seen for both peptides with excess of peptides over available zinc. PMID- 15536072 TI - The plasma membrane Q-SNARE syntaxin 2 enters the zymogen granule membrane during exocytosis in the pancreatic acinar cell. AB - During exocytosis in the pancreatic acinar cell, zymogen granules fuse directly with the apical plasma membrane and also with granules that have themselves fused with the plasma membrane. Together, these primary and secondary fusion events constitute the process of compound exocytosis. It has been suggested that the sequential nature of primary and secondary fusion is a consequence of the requirement for plasma membrane soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors, such as syntaxin 2, to enter the membrane of the primary fused granule. We have tested this possibility by determining the location of syntaxin 2 in unstimulated and stimulated pancreatic acini. Syntaxin 2 was imaged by confocal immunofluorescence microscopy. Fused granules were detected both through their filling with the aqueous dye lysine-fixable Texas Red dextran and through the decoration of their cytoplasmic surfaces with filamentous actin. In unstimulated cells, syntaxin 2 was exclusively present on the apical plasma membrane. In contrast, after stimulation, syntaxin 2 had moved into the membranes of fused granules, as judged by its location around dye-filled structures of 1-mum diameter that were coated with filamentous actin. At long times of stimulation (5 min), the majority (85%) of dye-filled granules were also positive for syntaxin 2. In contrast, at shorter times (1 min), more dye-filled granules (29%) were syntaxin 2-negative. We conclude that syntaxin 2 enters the membrane of a fused zymogen granule after the opening of the fusion pore, and we suggest that this movement might permit the onset of secondary fusion. PMID- 15536073 TI - The hyperglycemia-induced inflammatory response in adipocytes: the role of reactive oxygen species. AB - Hyperglycemia is a major independent risk factor for diabetic macrovascular disease. The consequences of exposure of endothelial cells to hyperglycemia are well established. However, little is known about how adipocytes respond to both acute as well as chronic exposure to physiological levels of hyperglycemia. Here, we analyze adipocytes exposed to hyperglycemia both in vitro as well as in vivo. Comparing cells differentiated at 4 mm to cells differentiated at 25 mm glucose (the standard differentiation protocol) reveals severe insulin resistance in cells exposed to 25 mm glucose. A global assessment of transcriptional changes shows an up-regulation of a number of mitochondrial proteins. Exposure to hyperglycemia is associated with a significant induction of reactive oxygen species (ROS), both in vitro as well as in vivo in adipocytes isolated from streptozotocin-treated hyperglycemic mice. Furthermore, hyperglycemia for a few hours in a clamped setting will trigger the induction of a pro-inflammatory response in adipose tissue from rats that can effectively be reduced by co infusion of N-acetylcysteine (NAC). ROS levels in 3T3-L1 adipocytes can be reduced significantly with pharmacological agents that lower the mitochondrial membrane potential, or by overexpression of uncoupling protein 1 or superoxide dismutase. In parallel with ROS, interleukin-6 secretion from adipocytes is significantly reduced. On the other hand, treatments that lead to a hyperpolarization of the mitochondrial membrane, such as overexpression of the mitochondrial dicarboxylate carrier result in increased ROS formation and decreased insulin sensitivity, even under normoglycemic conditions. Combined, these results highlight the importance ROS production in adipocytes and the associated insulin resistance and inflammatory response. PMID- 15536074 TI - Inhibition of integrin-mediated cell adhesion but not directional cell migration requires catalytic activity of EphB3 receptor tyrosine kinase. Role of Rho family small GTPases. AB - Genetic studies have shown that Eph receptor tyrosine kinases have both kinase dependent and kinase-independent functions through incompletely understood mechanisms. We report here that ephrin-B1 stimulation of endogenous EphB kinases in LS174T colorectal epithelial cells inhibited integrin-mediated adhesion and HGF/SF-induced directional cell migration. Using 293 cells stably transfected with wild type (WT)- or kinase-deficient (KD-EphB3), we found that inhibition of integrin-mediated cell adhesion and induction of cell rounding was kinase dependent. Unexpectedly, in two independent assays, both KD- and WT-EphB3 significantly inhibited directional cell migration. Upon ephrin-B1 stimulation, the activities of Rac1 and Cdc42 were reduced in both WT- and KD-EphB3-expressing cells that were induced to migrate. Pharmacological evidence demonstrates that a relative increase in RhoA signaling as a result of decreased Rac1/Cdc42 activities contributes to the inhibitory effects. Furthermore, EphB3-mediated inhibitory effect on cell adhesion but not migration was abolished by the integrin activating antibodies, suggesting that the inhibition of cell migration is not because of down-regulation of integrin function. These results uncover a differential requirement for EphB3 catalytic activity in the regulation of cell adhesion and migration, and suggest that while catalytic activity of EphB3 is required for inhibition of integrin-mediated cell adhesion, a distinct signaling pathway to Rho GTPases shared by WT- and KD-EphB3 receptor mediates inhibition of directional cell migration. PMID- 15536075 TI - A novel peptide isolated from a phage display peptide library with trastuzumab can mimic antigen epitope of HER-2. AB - Trastuzumab, a humanized antibody to HER-2, has been shown to be effective in the treatment of breast cancer in which HER-2 overexpression and metastasis occurs. In our search for an effective mimic epitope of HER-2 binding with trastuzumab and to develop HER-2 peptide vaccine, we screened a phage display 12-mer peptide library with trastuzumab as the target. A mimetic peptide (mimotope) H98 (LLGPYELWELSH) that could specifically recognize trastuzumab was isolated. The DNA encoding peptide H98 was cloned and expressed as the fusion protein GST-H98 in Escherichia coli BL21. The purified GST-H98 could specifically bind to trastuzumab and block the binding of trastuzumab to HER-2 protein. Moreover, H98 could significantly block the function of trastuzumab inhibiting the growth of cancer cells. Mice that were immunized with GST-H98 made specific antibody to H98 as well as to HER-2. In addition, T-cell proliferation occurred in mice immunized with GST-H98. Although no sequence homology was found between H98 and HER-2, through the use of structure analysis we were able to determine that peptide H98 contributed to a conformational epitope of HER-2. Furthermore, we determined that the last two amino acids at the C terminus, and the third together with the fourth amino acid at the N terminus of peptide H98 are critical to the binding of H98 to trastuzumab. As a result, we conclude that peptide H98 has potential for being developed as a HER-2 vaccine for biotherapy of cancer with HER-2 overexpression. PMID- 15536076 TI - cAMP has distinct acute and chronic effects on aquaporin-5 in lung epithelial cells. AB - Aquaporin-5 (AQP5) is present on the apical membrane of epithelial cells in various secretory glands as well as on the apical membrane of the airway epithelium, airway submucosal glands, and type 1 pneumocytes, where it can participate in respiratory tract water homeostasis. We examined the effects of cAMP on AQP5 distribution and abundance. When AQP5-expressing mouse lung epithelial cells were treated with cAMP or the beta-adrenergic agonist terbutaline, a biphasic AQP5 response was observed. Short term (minutes) exposure to cAMP produced internalization of AQP5 off of the membrane and a decrease in protein abundance. Both of these responses were blocked by inhibition of protein kinase A and the decrease in abundance was blocked by chloroquine, indicating lysosome-mediated degradation. Sustained cAMP exposure (hours) produced an increase in membrane localization and increased abundance; these effects were also blocked by protein kinase A inhibition. The beta-adrenergic agonist terbutaline produced changes in AQP5 abundance in mouse trachea and lung, consistent with our findings in cultured epithelial cells. Purified AQP5 protein was phosphorylated by protein kinase A but not protein kinase C or casein kinase II, and aquaporin-5 was phosphorylated in cultured cells after long term (but not short term) exposure to cAMP. These studies indicate that cAMP and beta adrenergic agonists produce distinct short and long term effects on AQP5 distribution and abundance that may contribute to regulation of lung water homeostasis. PMID- 15536077 TI - Bile acids induce a cationic current, depolarizing pancreatic acinar cells and increasing the intracellular Na+ concentration. AB - Biliary disease is a major cause of acute pancreatitis. In this study we investigated the electrophysiological effects of bile acids on pancreatic acinar cells. In perforated patch clamp experiments we found that taurolithocholic acid 3-sulfate depolarized pancreatic acinar cells. At low bile acid concentrations this occurred without rise in the cytosolic calcium concentration. Measurements of the intracellular Na(+) concentration with the fluorescent probe Sodium Green revealed a substantial increase upon application of the bile acid. We found that bile acids induce Ca(2+)-dependent and Ca(2+)-independent components of the Na(+) concentration increase. The Ca(2+)-independent component was resolved in conditions when the cytosolic Ca(2+) level was buffered with a high concentration of the calcium chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). The Ca(2+)-dependent component of intracellular Na(+) increase was clearly seen during stimulation with the calcium-releasing agonist acetylcholine. During acetylcholine-induced Ca(2+) oscillations the recovery of cytosolic Na(+) was much slower than the recovery of Ca(2+), creating a possibility for the summation of Na(+) transients. The bile-induced Ca(2+)-independent current was found to be carried primarily by Na(+) and K(+), with only small Ca(2+) and Cl(-) contributions. Measurable activation of such a cationic current could be produced by a very low concentration of taurolithocholic acid 3-sulfate (10 microm). This bile acid induced a cationic current even when applied in sodium- and bicarbonate free solution. Other bile acids, taurochenodeoxycholic acid, taurocholic acid, and bile itself also induced cationic currents. Bile-induced depolarization of acinar cells should have a profound effect on acinar fluid secretion and, consequently, on transport of secreted zymogens. PMID- 15536078 TI - Regulation of outside-in signaling in platelets by integrin-associated protein kinase C beta. AB - Studies with inhibitors have implicated protein kinase C (PKC) in the adhesive functions of integrin alpha(IIb)beta(3) in platelets, but the responsible PKC isoforms and mechanisms are unknown. Alpha(IIb)beta(3) interacts directly with tyrosine kinases c-Src and Syk. Therefore, we asked whether alpha(IIb)beta(3) might also interact with PKC. Of the several PKC isoforms expressed in platelets, only PKC beta co-immunoprecipitated with alpha(IIb)beta(3) in response to the interaction of platelets with soluble or immobilized fibrinogen. PKC beta recruitment to alpha(IIb)beta(3) was accompanied by a 9-fold increase in PKC activity in alpha(IIb)beta(3) immunoprecipitates. RACK1, an intracellular adapter for activated PKC beta, also co-immunoprecipitated with alpha(IIb)beta(3), but in this case, the interaction was constitutive. Broad spectrum PKC inhibitors blocked both PKC beta recruitment to alpha(IIb)beta(3) and the spread of platelets on fibrinogen. Similarly, mouse platelets that are genetically deficient in PKC beta spread poorly on fibrinogen, despite normal agonist-induced fibrinogen binding. In a Chinese hamster ovary cell model system, adhesion to fibrinogen caused green fluorescent protein-PKC beta I to associate with alpha(IIb)beta(3) and to co-localize with it at lamellipodial edges. These responses, as well as Chinese hamster ovary cell migration on fibrinogen, were blocked by the deletion of the beta(3) cytoplasmic tail or by co-expression of a RACK1 mutant incapable of binding to beta(3). These studies demonstrate that the interaction of alpha(IIb)beta(3) with activated PKC beta is regulated by integrin occupancy and can be mediated by RACK1 and that the interaction is required for platelet spreading triggered through alpha(IIb)beta(3). Furthermore, the studies extend the concept of alpha(IIb)beta(3) as a scaffold for multiple protein kinases that regulate the platelet actin cytoskeleton. PMID- 15536079 TI - Molecular dissection of interactions between components of the alternative pathway of complement and decay accelerating factor (CD55). AB - The complement regulatory protein decay accelerating factor (DAF; CD55), inhibits the alternative complement pathway by accelerating decay of the convertase enzymes formed by C3b and factor B. We show, using surface plasmon resonance, that in the absence of Mg(2+), DAF binds C3b, factor B, and the Bb subunit with low affinity (K(D), 14 +/- 0.1, 44 +/- 10, and 20 +/- 7 microm, respectively). In the presence of Mg(2+), DAF bound Bb or the von Willebrand factor type A subunit of Bb with higher affinities (K(D), 1.3 +/- 0.5 and 2.2 +/- 0.1 microm, respectively). Interaction with the proenzyme C3bB was investigated by flowing factor B across a C3b-coated surface in the absence of factor D. The dissociation rate was dependent on the time of incubation, suggesting that a time-dependent conformational transition stabilized the C3b-factor B interaction. Activation by factor D (forming C3bBb) increased the complex half-life; however, the enzyme became susceptible to rapid decay by DAF, unlike the proenzyme, which was unaffected. A convertase assembled with cobra venom factor and Bb was decayed by DAF, albeit far less efficiently than C3bBb. DAF did not bind cobra venom factor, implying that Bb decay is accelerated, at least in part, through DAF binding of this subunit. It is likely that DAF binds the complex with higher affinity/avidity, promoting a conformational change in either or both subunits accelerating decay. Such analysis of component and regulator interactions will inform our understanding of inhibitory mechanisms and the ways in which regulatory proteins cooperate to control the complement cascade. PMID- 15536080 TI - Molecular regulation of membrane resealing in 3T3 fibroblasts. AB - Membrane resealing in mammalian cells after injury depends on Ca(2+)-dependent fusion of intracellular vesicles with the plasma membrane. When cells are wounded twice, the subsequent resealing is generally faster. Physiological and biochemical studies have shown the initiation of two different repair signaling pathways, which are termed facilitated and potentiated responses. The facilitated response is dependent on the generation and recruitment of new vesicles, whereas the potentiated response is not. Here, we report that the two responses can be differentially defined molecularly. Using recombinant fragments of synaptobrevin 2 and synaptotagmin C2 domains we were able to dissociate the molecular requirements of vesicle exocytosis for initial membrane resealing and the facilitated and potentiated responses. The initial resealing response was blocked by fragments of synaptobrevin-2 and the C2B domain of synaptotagmin VII. Both the facilitated and potentiated responses were also blocked by the C2B domain of synaptotagmin VII. Although the initial resealing response was not blocked by the C2AB domain of synaptotagmin I or the C2A domain of synaptotagmin VII, recruitment of new vesicles for the facilitated response was inhibited. We also used Ca2+ binding mutant studies to show that the effects of synaptotagmins on membrane resealing are Ca(2+)-dependent. The pattern of inhibition by synaptotagmin C2 fragments that we observed cannot be used to specify a vesicle compartment, such as lysosomes, in membrane repair. PMID- 15536081 TI - Calcium-binding crystallins from Yersinia pestis. Characterization of two single betagamma-crystallin domains of a putative exported protein. AB - Betagamma-crystallin is a superfamily with diverse members from vertebrate lens to microbes. However, not many members have been identified and studied. Here, we report the identification of a putative exported protein from Yersinia pestis as a member of the betagamma-crystallin superfamily. Even though calcium has been known to play an important role in the physiology and virulence of the Yersinia genus, calcium-binding proteins have not yet been identified. We have studied the calcium-binding properties of two of the three crystallin domains present in this putative exported protein designated "Yersinia crystallin." These two domains (D1 and D2) have unique AA and BB types of arrangement of their Greek key motifs unlike the domains of other members of the betagamma-crystallin superfamily, which are either AB or BA types. These domains bind two calcium ions with low and high affinity-binding sites. We showed their calcium-binding properties using various probes for calcium and the effect of calcium on their secondary and tertiary structures. Although both domains bind calcium, D1 underwent drastic changes in secondary and tertiary structure and hydrodynamic volume upon calcium binding. Domain D1, which is intrinsically unstructured in the apo form, requires calcium for the typical betagamma-crystallin fold. Calcium exerted an extrinsic stabilization effect on domain D1 but not on D2, which is also largely unstructured. We suggest that this protein might be involved in calcium-dependent processes, such as stress response or physiology in the Yersinia genus, similar to its microbial relatives and mammalian lens crystallins. PMID- 15536082 TI - Human polyserase-2, a novel enzyme with three tandem serine protease domains in a single polypeptide chain. AB - We have cloned a human cDNA encoding a new serine protease that has been called polyserase-2 (polyserine protease-2) because it is the second identified human enzyme with several tandem serine protease domains in its amino acid sequence. The first serine protease domain contains all characteristic features of these enzymes, whereas the second and third domains lack one residue of the catalytic triad of serine proteases and are predicted to be catalytically inactive. This complex domain organization is also present in the sequences of mouse and rat polyserase-2 and resembles that of polyserase-1, which also contains three serine protease domains in its amino acid sequence. However, polyserase-2 lacks additional domains present in polyserase-1, including a type II transmembrane motif and a low-density lipoprotein receptor A module. Enzymatic analysis demonstrated that both full-length polyserase-2 and its first serine protease domain hydrolyzed synthetic peptides used for assaying serine proteases. Nevertheless, the activity of the isolated domain was greater than that of the entire protein, suggesting that the two catalytically inactive serine protease domains of polyserase-2 may modulate the activity of the first domain. Northern blot analysis showed that polyserase-2 is expressed in fetal kidney; adult skeletal muscle, liver, placenta, prostate, and heart; and tumor cell lines derived from lung and colon adenocarcinomas. Finally, analysis of post translational processing mechanisms of polyserase-2 revealed that, contrary to those affecting to the membrane-bound polyserase-1, this novel polyprotein is a secreted enzyme whose three protease domains remain as an integral part of a single polypeptide chain. PMID- 15536083 TI - CIL-102 interacts with microtubule polymerization and causes mitotic arrest following apoptosis in the human prostate cancer PC-3 cell line. AB - There have been no therapeutic agents that provide a survival advantage in hormone-refractory prostate cancer. Recently, the Food and Drug Administration approved docetaxel combined with prednisone for the treatment of patients with advanced metastatic prostate cancer, and it does show a survival benefit. Hence, anti-microtubule drugs might be of benefit in chemotherapy of hormone-refractory prostate cancer. We used metastatic hormone-refractory prostate cancer PC-3 cells to investigate potential molecular mechanisms for CIL-102, a semisynthetic alkaloid derivative. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide and sulforhodamine B assays indicated that CIL-102 inhibits cell growth dose-dependently. Immunofluorescence microscopy and in vitro tubulin assembly assays indicated that CIL-102 binds to tubulin and disrupts microtubule organization. Flow cytometry showed that CIL-102 causes cells to accumulate in G(2)/M phase and sub-G(0)/G(1) phase. CIL-102-induced apoptosis was also characterized by immunofluorescence microscopy. Western blotting and kinase assays showed that CIL-102 exposure induced up-regulation of cyclin B1 and p34(cdc2) kinase activity and olomoucine, a p34(cdc2) inhibitor, profoundly reduced the number of cells accumulated in mitotic phase. Moreover, Bcl-2 phosphorylation, Cdc25C phosphorylation, and survivin expression were increased. CIL-102-induced apoptosis was associated with activation of caspase-3, but a noncaspase pathway may also be involved, since benzyloxycarbonyl-VAD-fluoromethyl ketone, a pancaspase inhibitor, only partially inhibited the apoptosis, and apoptosis-inducing factor was translocated from mitochondria to cytosol. We conclude that CIL-102 induces mitotic arrest and apoptosis by binding to tubulin and inhibiting tubulin polymerization. CIL-102 causes mitotic arrest, at least partly, by modulating cyclin-dependent kinases and then apoptosis executed by caspase and noncaspase pathways. PMID- 15536085 TI - Epitope mapping of a monoclonal antibody specific to bovine dry milk: involvement of residues 66-76 of strand D in thermal denatured beta-lactoglobulin. AB - beta-Lactoglobulin (beta-LG) is a bovine milk protein sensitive to thermal denaturation. Previously, we demonstrated that such structural change can be detected by a monoclonal antibody (mAb) specific to denatured beta-LG. In the present study, we show a dramatic increase in beta-LG immunoreactivity when heating raw milk between 70 and 80 degrees C. To map out the specific epitope of beta-LG recognized by this mAb, we used a combined strategy including tryptic and CNBr fragments, chemical modifications (acetylation and carboxymethylation), peptide array containing in situ synthesized peptides, and a synthetic soluble peptide for immunoassays. The antigenic determinant we defined was exactly located within the D strand (residues 66-76) of beta-LG. Circular dichroic spectral analysis shows that carboxymethylation on beta-LG not only resulted in a substantial loss of beta-configuration but also exerted a 10 times increase in immunoreactivity as compared with heated beta-LG. The result suggests that a further disordered structure occurred in beta-LG and thus rendered the mAb recognition. Mutations on each charged residue (three Lys and one Glu) revealed that Lys-69 and Glu-74 were extremely essential in maintaining the antigenic structure. We also show an inverse relationship between the immunoreactivity in heated beta-LG and its binding to retinol or palmitic acid. Most interestingly, pH 9-10, which neutralizes the Lys groups of beta-LG, not only reduced its immunoreactivity but also its binding to palmitic acid implicating a role of Lys 69. Taken together, we concluded that strand D of beta-LG participated in the thermal denaturation between 70 and 80 degrees C and the binding to retinol and palmitic acid. The antigenic and biochemical roles of mAb specific to D strand are discussed in detail. PMID- 15536084 TI - Molecular basis for a direct interaction between the Syk protein-tyrosine kinase and phosphoinositide 3-kinase. AB - After engagement of the B cell receptor for antigen, the Syk protein-tyrosine kinase becomes phosphorylated on multiple tyrosines, some of which serve as docking sites for downstream effectors with SH2 or other phosphotyrosine binding domains. The most frequently identified binding partner for catalytically active Syk identified in a yeast two-hybrid screen was the p85 regulatory subunit of phosphoinositide 3-kinase. The C-terminal SH2 domain of p85 was sufficient for mediating an interaction with tyrosine-phosphorylated Syk. Interestingly, this domain interacted with Syk at phosphotyrosine 317, a site phosphorylated in trans by the Src family kinase, Lyn, and identified previously as a binding site for c Cbl. This site interacted preferentially with the p85 C-terminal SH2 domain compared with the c-Cbl tyrosine kinase binding domain. Molecular modeling studies showed a good fit between the p85 SH2 domain and a peptide containing phosphotyrosine 317. Tyr-317 was found to be essential for Syk to support phagocytosis mediated by FcgammaRIIA receptors expressed in a heterologous system. These studies establish a new type of p85 binding site that can exist on proteins that serve as substrates for Src family kinases and provide a molecular explanation for observations on direct interactions between Syk and phosphoinositide 3-kinase. PMID- 15536086 TI - RGS12 interacts with the SNARE-binding region of the Cav2.2 calcium channel. AB - Activation of GABAB receptors in chick dorsal root ganglion (DRG) neurons inhibits the Cav2.2 calcium channel in both a voltage-dependent and voltage independent manner. The voltage-independent inhibition requires activation of a tyrosine kinase that phosphorylates the alpha1 subunit of the channel and thereby recruits RGS12, a member of the "regulator of G protein signaling" (RGS) proteins. Here we report that RGS12 binds to the SNARE-binding or "synprint" region (amino acids 726-985) in loop II-III of the calcium channel alpha1 subunit. A recombinant protein encompassing the N-terminal PTB domain of RGS12 binds to the synprint region in protein overlay and surface plasmon resonance binding assays; this interaction is dependent on tyrosine phosphorylation and yet is within a sequence that differs from the canonical NPXY motif targeted by other PTB domains. In electrophysiological experiments, microinjection of DRG neurons with synprint-derived peptides containing the tyrosine residue Tyr-804 altered the rate of desensitization of neurotransmitter-mediated inhibition of the Cav2.2 calcium channel, whereas peptides centered about a second tyrosine residue, Tyr 815, were without effect. RGS12 from a DRG neuron lysate was precipitated using synprint peptides containing phosphorylated Tyr-804. The high degree of conservation of Tyr-804 in the SNARE-binding region of Cav2.1 and Cav2.2 calcium channels suggests that this region, in addition to the binding of SNARE proteins, is also important for determining the time course of the modulation of calcium current via tyrosine phosphorylation. PMID- 15536087 TI - Translational control of beta2-adrenergic receptor mRNA by T-cell-restricted intracellular antigen-related protein. AB - Cellular expression of the beta(2)-adrenergic receptor (beta(2)-AR) is suppressed at the translational level by 3'-untranslated region (UTR) sequences. To test the possible role of 3'-UTR-binding proteins in translational suppression of beta(2) AR mRNA, we expressed the full-length 3'-UTR or the adenylate/uridylate-rich (A+U rich element (ARE)) RNA from the 3'-UTR sequences of beta(2)-AR in cell lines that endogenously express this receptor. Reversal of beta(2)-adrenergic receptor translational repression by retroviral expression of 3'-UTR sequences suggested that ARE RNA-binding proteins are involved in translational suppression of beta(2)-adrenergic receptor expression. Using a 20-nucleotide ARE RNA from the receptor 3'-UTR as an affinity ligand, we purified the proteins that bind to these sequences. T-cell-restricted intracellular antigen-related protein (TIAR) was one of the strongly bound proteins identified by this method. UV-catalyzed cross-linking experiments using in vitro transcribed 3'-UTR RNA and glutathione S transferase-TIAR demonstrated multiple binding sites for this protein on beta(2) AR 3'-UTR sequences. The distal 340-nucleotide region of the 3'-UTR was identified as a target RNA motif for TIAR binding by both RNA gel shift analysis and immunoprecipitation experiments. Overexpression of TIAR resulted in suppression of receptor protein synthesis and a significant shift in endogenously expressed beta(2)-AR mRNA toward low molecular weight fractions in sucrose gradient polysome fractionation. Taken together, our results provide the first evidence for translational control of beta(2)-AR mRNA by TIAR. PMID- 15536088 TI - Ubiquitination of the peroxisomal targeting signal type 1 receptor, Pex5p, suggests the presence of a quality control mechanism during peroxisomal matrix protein import. AB - PEX genes encode proteins (peroxins) that are required for the biogenesis of peroxisomes. One of these peroxins, Pex5p, is the receptor for matrix proteins with a type 1 peroxisomal targeting signal (PTS1), which shuttles newly synthesized proteins from the cytosol into the peroxisome matrix. We observed that in various Saccharomyces cerevisiae pex mutants disturbed in the early stages of PTS1 import, the steady-state levels of Pex5p are enhanced relative to wild type controls. Furthermore, we identified ubiquitinated forms of Pex5p in deletion mutants of those PEX genes that have been implicated in recycling of Pex5p from the peroxisomal membrane into the cytosol. Pex5p ubiquitination required the presence of the ubiquitin-conjugating enzyme Ubc4p and the peroxins that are required during early stages of PTS1 protein import. Finally, we provide evidence that the proteasome is involved in the turnover of Pex5p in wild type yeast cells, a process that requires Ubc4p and occurs at the peroxisomal membrane. Our data suggest that during receptor recycling a portion of Pex5p becomes ubiquitinated and degraded by the proteasome. We propose that this process represents a conserved quality control mechanism in peroxisome biogenesis. PMID- 15536089 TI - CTP: Phosphocholine cytidylyltransferase: paving the way from gene to membrane. PMID- 15536090 TI - The Ca2+ channel alpha2delta-1 subunit determines Ca2+ current kinetics in skeletal muscle but not targeting of alpha1S or excitation-contraction coupling. AB - Auxiliary channel subunits regulate membrane expression and modulate current properties of voltage-activated Ca(2+) channels and thus are involved in numerous important cell functions, including muscle contraction. Whereas the importance of the alpha(1S), beta(1a), and gamma Ca(2+) channel subunits in skeletal muscle has been determined by using null-mutant mice, the role of the alpha(2)delta-1 subunit in skeletal muscle is still elusive. We addressed this question by small interfering RNA silencing of alpha(2)delta-1 in reconstituted dysgenic (alpha(1S) null) myotubes and in BC3H1 skeletal muscle cells. Immunofluorescence labeling of the alpha(1S) and alpha(2)delta-1 subunits and whole cell patch clamp recordings demonstrated that triad targeting and functional expression of the skeletal muscle Ca(2+) channel were not compromised by the depletion of the alpha(2)delta 1 subunit. The amplitudes and voltage dependences of L-type Ca(2+) currents and of the depolarization-induced Ca(2+) transients were identical in control and in alpha(2)delta-1-depleted muscle cells. However, alpha(2)delta-1 depletion significantly accelerated the current kinetics, most likely by the conversion of slowly activating into fast activating Ca(2+) channels. Reverse transcription-PCR analysis indicated that alpha(2)delta-1 is the exclusive isoform expressed in differentiated BC3H1 cells and that depletion of alpha(2)delta-1 was not compensated by the up-regulation of any other alpha(2)delta isoform. Thus, in skeletal muscle the Ca(2+) channel alpha(2)delta-1 subunit functions as a major determinant of the characteristic slow L-type Ca(2+) current kinetics. However, this subunit is not essential for targeting of Ca(2+) channels or for their primary physiological role in activating skeletal muscle excitation-contraction coupling. PMID- 15536091 TI - Fyn phosphorylates human MAP-2c on tyrosine 67. AB - The Src homology 3 (SH3) domain of Fyn binds to a conserved PXXP motif on microtubule-associated protein-2. Co-transfections into COS7 cells and in vitro kinase assays performed with Fyn and wild-type, or mutant MAP-2c, determined that Fyn phosphorylated MAP-2c on tyrosine 67. The phosphorylation generated a consensus sequence for the binding of the SH2 domain of Grb2 (pYSN). Pull-down assays with SH2-Grb2 from human fetal brain homogenates, and co immunoprecipitation of Grb2 and MAP-2 confirmed the interaction in vivo, and demonstrated that MAP-2c is tyrosine-phosphorylated in human fetal brain. Filter overlay assays confirmed that the SH2 domain of Grb2 binds to human MAP-2c following incubation with active Fyn. Enzyme-linked immunosorbent assays confirmed the interaction between the SH2 domain of Grb2 and a tyrosine phosphorylated MAP-2 peptide spanning the pY(67)SN motif. Thus, MAP-2c can directly recruit multiple signaling proteins important for central nervous system development. PMID- 15536092 TI - An intermediate form of ADP-F-actin. AB - With yeast actin, contrary to other actins, filament formation, ATP hydrolysis, and Pi release are concurrent at low actin concentrations, the condition usually employed to assess actin polymerization. This observation leads to a question concerning the conformation of the filament barbed end that might be recognized by specific actin-binding proteins. To try to detect possible new actin polymer conformations that might be intermediate in the pathway leading to mature F actin, we monitored the change in intrinsic tryptophan fluorescence of yeast and muscle actins polymerized at pH 6 to accelerate the rate of filament formation. This allowed temporal resolution of the Pi release process from the slower process of polymerization. With both actins, we detected a biphasic instead of the usual monophasic fluorescence change, a rapid decrease that tracks with filament formation followed by a slower rebound (the second phase). This second phase postpolymerization conformational change requires Pi release and occurs nearly coincident with its release. The addition of Pi causes this second phase response to disappear, and the inclusion of Pi during polymerization prevents its appearance. At pH 7.5, with higher yeast actin concentrations to accelerate polymerization, a two-phase fluorescence change is also observed. In this case, the second phase change lags substantially behind Pi release. Pi release could also be resolved from polymer formation. V159N yeast actin, hypothesized previously as remaining in a postpolymerization ATP-like state, exhibits the same two-phase intrinsic tryptophan fluorescence behavior as wild-type yeast actin. Together, these observations demonstrate the presence of an intermediate filament state between ADP-Pi and mature ADP-F-actin. PMID- 15536093 TI - Syncope Evaluation in the Emergency Department Study (SEEDS): a multidisciplinary approach to syncope management. AB - BACKGROUND: The primary aim and central hypothesis of the study are that a designated syncope unit in the emergency department improves diagnostic yield and reduces hospital admission for patients with syncope who are at intermediate risk for an adverse cardiovascular outcome. METHODS AND RESULTS: In this prospective, randomized, single-center study, patients were randomly allocated to 2 treatment arms: syncope unit evaluation and standard care. The 2 groups were compared with chi2 test for independence of categorical variables. Wilcoxon rank sum test was used for continuous variables. Survival was estimated with the Kaplan-Meier method. One hundred three consecutive patients (53 women; mean age 64+/-17 years) entered the study. Fifty-one patients were randomized to the syncope unit. For the syncope unit and standard care patients, the presumptive diagnosis was established in 34 (67%) and 5 (10%) patients (P<0.001), respectively, hospital admission was required for 22 (43%) and 51 (98%) patients (P<0.001), and total patient-hospital days were reduced from 140 to 64. Actuarial survival was 97% and 90% (P=0.30), and survival free from recurrent syncope was 88% and 89% (P=0.72) at 2 years for the syncope unit and standard care groups, respectively. CONCLUSIONS: The novel syncope unit designed for this study significantly improved diagnostic yield in the emergency department and reduced hospital admission and total length of hospital stay without affecting recurrent syncope and all-cause mortality among intermediate-risk patients. Observations from the present study provide benchmark data for improving patient care and effectively utilizing healthcare resources. PMID- 15536095 TI - When cost is an adverse drug effect, patients cut corners and risk health. PMID- 15536101 TI - Glycoprotein IIb/IIIa inhibition. PMID- 15536102 TI - Glycoprotein IIb/IIIa inhibition. PMID- 15536103 TI - Health care delivery in the Texas prison system. PMID- 15536104 TI - Health care delivery in the Texas prison system. PMID- 15536105 TI - Small intestinal bacterial overgrowth and irritable bowel syndrome. PMID- 15536106 TI - Augmented-reality-assisted laparoscopic adrenalectomy. PMID- 15536107 TI - Stimulants and attention-deficit/hyperactivity disorder. PMID- 15536108 TI - Effect of antihypertensive agents on cardiovascular events in patients with coronary disease and normal blood pressure: the CAMELOT study: a randomized controlled trial. AB - CONTEXT: The effect of antihypertensive drugs on cardiovascular events in patients with coronary artery disease (CAD) and normal blood pressure remains uncertain. OBJECTIVE: To compare the effects of amlodipine or enalapril vs placebo on cardiovascular events in patients with CAD. DESIGN, SETTING, AND PARTICIPANTS: Double-blind, randomized, multicenter, 24-month trial (enrollment April 1999-April 2002) comparing amlodipine or enalapril with placebo in 1991 patients with angiographically documented CAD (>20% stenosis by coronary angiography) and diastolic blood pressure <100 mm Hg. A substudy of 274 patients measured atherosclerosis progression by intravascular ultrasound (IVUS). INTERVENTIONS: Patients were randomized to receive amlodipine, 10 mg; enalapril, 20 mg; or placebo. IVUS was performed at baseline and study completion. MAIN OUTCOME MEASURES: The primary efficacy parameter was incidence of cardiovascular events for amlodipine vs placebo. Other outcomes included comparisons of amlodipine vs enalapril and enalapril vs placebo. Events included cardiovascular death, nonfatal myocardial infarction, resuscitated cardiac arrest, coronary revascularization, hospitalization for angina pectoris, hospitalization for congestive heart failure, fatal or nonfatal stroke or transient ischemic attack, and new diagnosis of peripheral vascular disease. The IVUS end point was change in percent atheroma volume. RESULTS: Baseline blood pressure averaged 129/78 mm Hg for all patients; it increased by 0.7/0.6 mm Hg in the placebo group and decreased by 4.8/2.5 mm Hg and 4.9/2.4 mm Hg in the amlodipine and enalapril groups, respectively (P<.001 for both vs placebo). Cardiovascular events occurred in 151 (23.1%) placebo-treated patients, in 110 (16.6%) amlodipine-treated patients (hazard ratio [HR], 0.69; 95% CI, 0.54-0.88 [P = .003]), and in 136 (20.2%) enalapril-treated patients (HR, 0.85; 95% CI, 0.67-1.07 [P = .16]. Primary end point comparison for enalapril vs amlodipine was not significant (HR, 0.81; 95% CI, 0.63-1.04 [P = .10]). The IVUS substudy showed a trend toward less progression of atherosclerosis in the amlodipine group vs placebo (P = .12), with significantly less progression in the subgroup with systolic blood pressures greater than the mean (P = .02). Compared with baseline, IVUS showed progression in the placebo group (P<.001), a trend toward progression in the enalapril group (P = .08), and no progression in the amlodipine group (P = .31). For the amlodipine group, correlation between blood pressure reduction and progression was r = 0.19, P = .07. CONCLUSIONS: Administration of amlodipine to patients with CAD and normal blood pressure resulted in reduced adverse cardiovascular events. Directionally similar, but smaller and nonsignificant, treatment effects were observed with enalapril. For amlodipine, IVUS showed evidence of slowing of atherosclerosis progression. PMID- 15536109 TI - Metabolic effects of carvedilol vs metoprolol in patients with type 2 diabetes mellitus and hypertension: a randomized controlled trial. AB - CONTEXT: Beta-blockers have been shown to decrease cardiovascular risk in patients with hypertension and type 2 diabetes mellitus (DM); however, some components of the metabolic syndrome are worsened by some beta-blockers. OBJECTIVE: To compare the effects of beta-blockers with different pharmacological profiles on glycemic and metabolic control in participants with DM and hypertension receiving renin-angiotensin system (RAS) blockade, in the context of cardiovascular risk factors. DESIGN, SETTING, AND PARTICIPANTS: A randomized, double-blind, parallel-group trial (The Glycemic Effects in Diabetes Mellitus: Carvedilol-Metoprolol Comparison in Hypertensives [GEMINI]) conducted between June 1, 2001, and April 6, 2004, at 205 US sites that compared the effects of carvedilol and metoprolol tartrate on glycemic control. The 1235 participants were aged 36 to 85 years with hypertension (>130/80 mm Hg) and type 2 DM (glycosylated hemoglobin [HbA1c], 6.5%-8.5%) and were receiving RAS blockers. Participants were followed up for 35 weeks. INTERVENTIONS: Participants were randomized to receive a 6.25- to 25-mg dose of carvedilol (n = 498) or 50- to 200 mg dose of metoprolol tartrate (n = 737), each twice daily. Open-label hydrochlorothiazide and a dihydropyridine calcium antagonist were added, if needed, to achieve blood pressure target. MAIN OUTCOME MEASURES: Difference between groups in mean change from baseline HbA1c following 5 months of maintenance therapy. Additional prespecified comparisons included change from baseline HbA1c in individual treatment groups, treatment effect on insulin sensitivity, and microalbuminuria. RESULTS: The 2 groups differed in mean change in HbA1c from baseline (0.13%; 95% confidence interval [CI], -0.22% to -0.04%; P = .004; modified intention-to-treat analysis). The mean (SD) HbA1c increased with metoprolol (0.15% [0.04%]; P<.001) but not carvedilol (0.02% [0.04%]; P = .65). Insulin sensitivity improved with carvedilol (-9.1%; P = .004) but not metoprolol (-2.0%; P = .48); the between-group difference was -7.2% (95% CI, -13.8% to 0.2%; P = .004). Blood pressure was similar between groups. Progression to microalbuminuria was less frequent with carvedilol than with metoprolol (6.4% vs 10.3%; odds ratio, 0.60; 95% CI, 0.36-0.97; P = .04). CONCLUSIONS: Both beta blockers were well tolerated; use of carvedilol in the presence of RAS blockade did not affect glycemic control and improved some components of the metabolic syndrome relative to metoprolol in participants with DM and hypertension. The effects of the 2 beta-blockers on clinical outcomes need to be compared in long term clinical trials. PMID- 15536110 TI - The metabolic syndrome, inflammation, and risk of cognitive decline. AB - CONTEXT: Several studies have reported an association between the metabolic syndrome and cardiovascular disease. Despite an increasing awareness that cardiovascular risk factors increase risk of cognitive decline and dementia, there are few data on the metabolic syndrome and cognition. OBJECTIVE: To determine if the metabolic syndrome is a risk factor for cognitive decline and if this association is modified by inflammation. DESIGN AND SETTING: A 5-year prospective observational study conducted from 1997 to 2002 at community clinics at 2 sites. PARTICIPANTS: A total of 2632 black and white elders (mean age, 74 years). MAIN OUTCOME MEASURES: Association of the metabolic syndrome (measured using National Cholesterol Education Program guidelines) and high inflammation (defined as above median serum level of interleukin 6 and C-reactive protein) with change in cognition (Modified Mini-Mental State Examination [3MS]) at 3 and 5 years. Cognitive impairment was defined as at least a 5-point decline. RESULTS: Compared with those without the metabolic syndrome (n = 1616), elders with the metabolic syndrome (n = 1016) were more likely to have cognitive impairment (26% vs 21%, multivariate adjusted relative risk [RR], 1.20; 95% confidence interval [CI], 1.02-1.41). There was a statistically significant interaction with inflammation and the metabolic syndrome (P = .03) on cognitive impairment. After stratifying for inflammation, those with the metabolic syndrome and high inflammation (n = 348) had an increased likelihood of cognitive impairment compared with those without the metabolic syndrome (multivariate adjusted RR, 1.66; 95% CI, 1.19-2.32). Those with the metabolic syndrome and low inflammation (n = 668) did not exhibit an increased likelihood of impairment (multivariate adjusted RR, 1.08; 95% CI, 0.89-1.30). Stratified multivariate random-effects models demonstrated that participants with the metabolic syndrome and high inflammation had greater 4-year decline on 3MS (P = .04) compared with those without the metabolic syndrome, whereas those with the metabolic syndrome and low inflammation did not (P = .44). CONCLUSION: These findings support the hypothesis that the metabolic syndrome contributes to cognitive impairment in elders, but primarily in those with high level of inflammation. PMID- 15536111 TI - Effect of DHEA on abdominal fat and insulin action in elderly women and men: a randomized controlled trial. AB - CONTEXT: Dehydroepiandrosterone (DHEA) administration has been shown to reduce accumulation of abdominal visceral fat and protect against insulin resistance in laboratory animals, but it is not known whether DHEA decreases abdominal obesity in humans. DHEA is widely available as a dietary supplement without a prescription. OBJECTIVE: To determine whether DHEA replacement therapy decreases abdominal fat and improves insulin action in elderly persons. DESIGN AND SETTING: Randomized, double-blind, placebo-controlled trial conducted in a US university based research center from June 2001 to February 2004. PARTICIPANTS: Fifty-six elderly persons (28 women and 28 men aged 71 [range, 65-78] years) with age related decrease in DHEA level. INTERVENTION: Participants were randomly assigned to receive 50 mg/d of DHEA or matching placebo for 6 months. MAIN OUTCOME MEASURES: The primary outcome measures were 6-month change in visceral and subcutaneous abdominal fat measured by magnetic resonance imaging and glucose and insulin responses to an oral glucose tolerance test (OGTT). RESULTS: Of the 56 men and women enrolled, 52 underwent follow-up evaluations. Compliance with the intervention was 97% in the DHEA group and 95% in the placebo group. Based on intention-to-treat analyses, DHEA therapy compared with placebo induced significant decreases in visceral fat area (-13 cm2 vs +3 cm2, respectively; P = .001) and subcutaneous fat (-13 cm2 vs +2 cm2, P = .003). The insulin area under the curve (AUC) during the OGTT was significantly reduced after 6 months of DHEA therapy compared with placebo (-1119 muU/mL per 2 hours vs +818 muU/mL per 2 hours, P = .007). Despite the lower insulin levels, the glucose AUC was unchanged, resulting in a significant increase in an insulin sensitivity index in response to DHEA compared with placebo (+1.4 vs -0.7, P = .005). CONCLUSION: DHEA replacement could play a role in prevention and treatment of the metabolic syndrome associated with abdominal obesity. PMID- 15536112 TI - First-trimester placentation and the risk of antepartum stillbirth. AB - CONTEXT: Preterm birth and low birth weight are determined, at least in part, during the first trimester of pregnancy. However, it is unknown whether the risk of stillbirth is also determined during the first trimester. OBJECTIVE: To determine whether the risk of antepartum stillbirth varies in relation to circulating markers of placental function measured during the first trimester of pregnancy. DESIGN, SETTING, AND PARTICIPANTS: Multicenter, prospective cohort study (conducted in Scotland from 1998 through 2000) of 7934 women who had singleton births at or after 24 weeks' gestation, who had blood taken during the first 10 weeks after conception, and who were entered into national registries of births and perinatal deaths. MAIN OUTCOME MEASURES: Antepartum stillbirths and stillbirths due to specific causes. RESULTS: There were 8 stillbirths among the 400 women with levels of pregnancy-associated plasma protein A (PAPP-A) in the lowest fifth percentile compared with 17 among the remaining 7534 women (incidence rate per 10,000 women per week of gestation: 13.4 vs 1.4, respectively; hazard ratio [HR], 9.2 [95% confidence interval [CI], 4.0-21.4]; P<.001). When analyzed by cause of stillbirth, low level of PAPP-A was strongly associated with stillbirth due to placental dysfunction, defined as abruption or unexplained stillbirth associated with growth restriction (incidence rate: 11.7 vs 0.3, respectively; HR, 46.0 [95% CI, 11.9-178.0]; P<.001), but was not associated with other causes of stillbirth (incidence rate: 1.7 vs 1.1, respectively; HR, 1.4 [95% CI, 0.2-10.6]; P = .75). There was no relationship between having a low level of PAPP-A and maternal age, ethnicity, parity, height, body mass index, race, or marital status. Adjustment for maternal factors did not attenuate the strength of associations observed. There was no association between maternal circulating levels of the free beta subunit of human chorionic gonadotropin and stillbirth risk. CONCLUSION: The risk of stillbirth in late pregnancy may be determined by placental function in the first 10 weeks after conception. PMID- 15536113 TI - A 67-year-old man who e-mails his physician. PMID- 15536114 TI - A 46-year-old man with anxiety and nightmares after a motor vehicle collision, 2 years later. PMID- 15536115 TI - Human monocytic ehrlichiosis. AB - A 56-year-old man with a history of Wegener granulomatosis presented with 6 days of sinus congestion, fever, malaise, myalgias, episcleritis, and a morbilliform rash. An exacerbation of Wegener granulomatosis was the principal concern because of the frequency of flares in that disease. The patient developed acute renal failure, thrombocytopenia, transaminitis, and, finally, severe myocarditis that led to congestive heart failure. Additional history-taking and the evolution of his clinical features led to empirical treatment with doxycycline for human monocytic ehrlichiosis (HME). The diagnosis of HME was confirmed by both a polymerase chain reaction assay for Ehrlichia chaffeensis and by the demonstration of morulae within peripheral blood mononuclear cells. The patient improved promptly following institution of doxycycline, and his cardiac function returned to normal over a period of 4 months. PMID- 15536116 TI - What is the optimal blood pressure and drug therapy for patients with coronary artery disease? PMID- 15536117 TI - Health care information technology: progress and barriers. PMID- 15536118 TI - JAMA patient page. Ehrlichiosis. PMID- 15536119 TI - Scribble protein domain mapping reveals a multistep localization mechanism and domains necessary for establishing cortical polarity. AB - The Drosophila tumor suppressor protein Scribble is required for epithelial polarity, neuroblast polarity, neuroblast spindle asymmetry and limiting cell proliferation. It is a member of the newly described LAP protein family, containing 16 leucine rich repeats (LRRs), four PDZ domains and an extensive carboxyl-terminal (CT) domain. LRR and PDZ domains mediate protein-protein interactions, but little is know about their function within LAP family proteins. We have determined the role of the LRR, PDZ and CT domains for Scribble localization in neuroblasts and epithelia, and for Scribble function in neuroblasts. We found that the LRR and PDZ domains are both required for proper targeting of Scribble to septate junctions in epithelia; that the LRR domain is necessary and sufficient for cortical localization in mitotic neuroblasts, and that the PDZ2 domain is required for efficient cortical and apical localization of Scribble in neuroblasts. In addition, we show that the LRR domain is sufficient to target Miranda protein to the neuroblast cortex, but that LRR+PDZ will exclude Miranda from the cortex. Our results highlight the importance of both LRR and PDZ domains for the proper localization and function of Scribble in neuroblasts. PMID- 15536120 TI - Formation of multivesicular endosomes in Dictyostelium. AB - Multivesicular endosomes are present in virtually every eucaryotic cell, where they arise by intra-endosomal budding of the limiting endosomal membrane. Some genetic diseases such as Chediak-Higashi syndrome are characterized by enlarged membrane-filled endosomes. The same altered endosomal morphology can be observed in cells exposed to certain drugs, for example U18666A. The mechanisms involved are still poorly characterized, partially because this atypical budding event is particularly difficult to observe in mammalian cells. Taking advantage of the simplicity of the endosomal structure in Dictyostelium discoideum, we could visualize intraendosomal budding at the ultrastructural level. In this model organism, the drug U18666A was shown to stimulate intra-endosomal budding, while an inhibitor of PI 3-kinase activity was found to have no effect on this process. Inactivation of a Dictyostelium gene with similarity to the gene responsible for Chediak-Higashi syndrome did not alter the intra-endosomal budding or the accumulation of intra-endosomal membranes. Thus, although treatment with U18666A and inactivation of the Chediak-Higashi gene cause similar morphological defects in mammalian cells, observations in a different model reveal that their respective modes of action are different. PMID- 15536121 TI - betagamma subunits of heterotrimeric G-proteins contribute to Ca2+ release at fertilization in the sea urchin. AB - A cytoplasmic Ca2+ transient is required for egg activation at fertilization in all animals. The pathway leading to release of Ca2+ from the endoplasmic reticulum in echinoderms includes activation of a SRC homolog, followed by phospholipase Cgamma activation, and formation of inositol trisphosphate. However, the upstream activators or modulators of this signaling pathway are not known. We recently identified four Galpha subunits of heterotrimeric G-proteins present in the sea urchin egg, and here we find that activation of G-proteins of the Galphas and Galphaq type, but not Galphai or Galpha12 type, is required for normal Ca2+ dynamics at fertilization. The effects of these G-proteins are mediated by the Gbetagamma subunits, occur upstream of the cytoplasmic Ca2+ release, and influence both the amplitude of Ca2+ release and the duration of the lag phase. We propose integration of the G-protein input into the framework of signaling at sea urchin fertilization. PMID- 15536122 TI - Distribution of Can1p into stable domains reflects lateral protein segregation within the plasma membrane of living S. cerevisiae cells. AB - Recently, lipid-raft-based subdomains within the plasma membrane of living Saccharomyces cerevisiae cells were visualized using green fluorescent protein fusions, and non-overlapping subdomains containing either Pma1p or Can1p were distinguished. In this study, the long-term stability of the subdomains was investigated. Experiments with latrunculin A and nocodazole ruled out the involvement of cytoskeletal components in the stabilization of the subdomains. Also a putative role of the cell wall was excluded, because protoplasting of the cells changed neither the pattern nor the stability of the subdomains. By contrast, the expected inner dynamics of the membrane subdomains was documented by FRAP experiments. Finally, two other proteins were localized within the frame of the Can1p/Pma1p plasma-membrane partition. We show that Fur4p (another H+ symporter) and Sur7p (a protein of unknown function) occupy the Can1p subdomain. PMID- 15536123 TI - Requirements for the destruction of human Aurora-A. AB - The mitotic kinase Aurora A (Aur-A) is overexpressed in a high proportion of human tumors, often in the absence of gene amplification. In somatic cells, Aur-A protein levels fall following mitosis or upon overexpression of Cdh1, an activator of the ubiquitin ligase APC/C. Thus, mutations that reduce or block the rate of Aur-A destruction might also be expected to contribute to its oncogenic potential. Previous work had defined two short sequences of Xenopus Aur-A that are required for its Cdh1-inducible destruction in extracts of Xenopus eggs, an N terminal A box and a C-terminal D box, and a serine residue within the A box whose phosphorylation might inhibit destruction. Here, we show that these same sequences are required for the destruction of human Aur-A during mitotic exit and G1 in the somatic cell cycle. Expression of a dominant negative Cdh1 protein leads to accumulation of Aur-A, further indicating that the Cdh1-activated form of the APC/C is responsible for destruction of Aur-A during the somatic cell cycle in vivo. During the course of this work, we found some previously unsuspected problems in commonly used in vitro destruction assays, which can result in misleading results. Potentially confounding factors include: (i) the presence of D-box- and A-box-dependent destruction-promoting activities in the reticulocyte in vitro translation mix that is used to produce radiolabeled substrates for destruction assays; and (ii) the ability of green-fluorescent protein tags to reduce the destruction rate of Aur-A substantially. These findings have direct relevance for studies of Aur-A destruction itself, and for broader approaches that use in vitro translation products in screens for additional APC/C targets. PMID- 15536124 TI - Characterization of a novel ATR-dependent, Chk1-independent, intra-S-phase checkpoint that suppresses initiation of replication in Xenopus. AB - In most eukaryotes, replication origins fire asynchronously throughout S-phase according to a precise timing programme. When replication fork progression is inhibited, an intra-S-phase checkpoint is activated that blocks further origin firing and stabilizes existing replication forks to prevent them undergoing irreversible collapse. We show that chromatin incubated in Xenopus egg extracts displays a replication-timing programme in which firing of new replication origins during S phase depends on the continued activity of S-phase-inducing cyclin-dependent kinases. We also show that low concentrations of the DNA polymerase inhibitor aphidicolin, which only slightly slows replication-fork progression, strongly suppress further initiation events. This intra-S-phase checkpoint can be overcome by caffeine, an inhibitor of the ATM/ATR checkpoint kinases, or by neutralizing antibodies to ATR. However, depletion or inhibition of Chk1 did not abolish the checkpoint. We could detect no significant effect on fork stability when this intra-S-phase checkpoint was inhibited. Interestingly, although caffeine could prevent the checkpoint from being activated, it could not rescue replication if added after the timing programme would normally have been executed. This suggests that special mechanisms might be necessary to reverse the effects of the intra-S-phase checkpoint once it has acted on particular origins. PMID- 15536125 TI - Balance of proliferation and cell death between thyrocytes and myofibroblasts regulates thyroid fibrosis in granulomatous experimental autoimmune thyroiditis (G-EAT). AB - Severe granulomatous experimental autoimmune thyroiditis (G-EAT), which progresses to fibrosis, is induced in DBA/1 mice by adoptive transfer of mouse thyroglobulin-primed and -activated spleen cells. There is extensive destruction of thyrocytes and inflammatory cell infiltration including T cells, macrophages, neutrophils, and myofibroblasts (myofbs). Suppression of transforming growth factor-beta (TGF-beta) and deficiency of interferon-gamma (IFN-gamma) inhibit fibrosis, and inflammation eventually resolves. Thyrocyte destruction in wild type (WT) mice was a result of apoptosis, as many deoxynucleotide triphosphate nick-end labeling + apoptotic thyrocytes were present in these thyroids. The balance of apoptosis and proliferation between thyrocytes and myofbs may be important factors determining the outcome of inflammation to fibrosis versus resolution. Apoptosis and proliferation in thyrocytes versus myofbs were evaluated by dual-staining of cell-proliferating marker (Ki-67) or in situ cell death and cytokeratin or alpha-smooth muscle actin and were analyzed by confocal microscopy. Apoptotic and antiapoptotic molecules in G-EAT thyroids were detected by immunostaining. In WT thyroids, which develop fibrosis, only a few myofbs were apoptotic, and many myofbs were Ki-67+, Fas-associated death domain protein-like interleukin-1beta-converting enzyme-like inhibitory protein (FLIP)+, and Bcl-XL+. In contrast, proliferation was predominant on thyrocytes of IFN-gamma-/- mice or anti-TGF-beta-treated WT mice. These results indicate that apoptosis of inflammatory cells and regeneration of thyrocytes in IFN-gamma-/- mice and anti TGF-beta-treated WT mice may limit development of fibrosis, whereas excessive proliferation of myofbs and loss of thyrocytes in WT mice may contribute to fibrosis. PMID- 15536126 TI - Stages of change for physical activity in a community sample of adolescents. AB - The aims of the present study were to investigate (1) the proportion of adolescents in each of the stages of change, (2) the differences in psychosocial factors and in physical activity between the stages, and (3) the classification accuracy using several reference criteria. A random sample of 38 schools from the Flemish community in Belgium resulted in a sample of 5931 adolescents (mean age=14.8, range 12-18, 61% females). All adolescents completed a computerized questionnaire assessing demographic variables, physical activity, physical activity determinants and stages of change. Distribution across the stages was: precontemplation, n=684 (11.5%); contemplation, n=948 (16.0%); preparation, n=818 (13.8%); action, n=492 (8.3%); and maintenance, n=2989 (50.4%). Analyses revealed gender and age differences (P<0.001), with girls and older adolescents being more prevalent in the first three stages. Differences between the stages of change were related to higher levels of physical activity of different intensity and within different contexts (P<0.001), together with more favorable psychosocial determinants related to physical activity (P<0.001). No differences between stages were found for sedentary activities. Three subgroups of adolescents were identified based on psychosocial determinants and physical activity levels. About 71% of the adolescents placed themselves in a stage which was in accordance with their self-reported level of physical activity. PMID- 15536127 TI - Distinct roles of IL-12 and IL-15 in human natural killer cell activation by dendritic cells from secondary lymphoid organs. AB - Dendritic cells (DCs) are known to induce the growth and function of natural killer (NK) cells. Here, we address the capacity of DCs to interact with NK cells in human lymphoid organs and identify the role of specific DC-derived cytokines. We demonstrate that DCs colocalize with NK cells in the T cell areas of lymph nodes. In culture, DCs from either blood or spleen primarily stimulate the CD56(bright)CD16- NK cell subset, which is enriched in secondary lymphoid tissues. Blocking of IL-12 abolished DC-induced IFN-gamma secretion by NK cells, whereas membrane-bound IL-15 on DCs was essential for NK cell proliferation and survival. Maturation by CD40 ligation promoted the highest IL-15 surface presentation on DCs and led to the strongest NK cell proliferation induced by DCs. These results identify secondary lymphoid organs as a potential DC/NK cell interaction site and identify the distinct roles for DC-derived IL-12 and IL-15 in NK cell activation. PMID- 15536128 TI - Alpha-synuclein structures from fluorescence energy-transfer kinetics: implications for the role of the protein in Parkinson's disease. AB - Parkinson's disease is associated with the deposition and accumulation of alpha synuclein fibrils in the brain. A30P and A53T mutations have been linked to the early-onset familial disease state. Time-resolved tryptophan fluorescence energy transfer measurements have been used to probe the structures of pseudo-wild-type and mutant (A30P) alpha-synucleins at physiological pH (7.4), in acidic pH (4.4) solutions, and in the presence of SDS micelles, a membrane mimic. Fluorescent donor-energy acceptor (DA) distance distributions for six different tryptophan/3 nitro-tyrosine pairs reveal the presence of compact, intermediate, and extended conformations of the protein. CD spectra indicate that the protein develops substantial helical structure in the presence of SDS micelles. DA distributions show that micelles induce compaction in the N-terminal region and expansion of the acidic C terminus. In acidic solutions, there is an increased population of collapsed structures in the C-terminal region. Energy-transfer measurements demonstrate that the average DA distances for the W4-Y19 and Y19-W39 pairs are longer in one of the two disease-related mutants (A30P). PMID- 15536129 TI - Human cytomegalovirus encodes a highly specific RANTES decoy receptor. AB - The human cytomegalovirus pUL21.5 protein is a small, secreted glycoprotein whose mRNA is packaged into virions. We demonstrate that pUL21.5 protein is a soluble CC chemokine receptor that functions as a decoy to modulate the host immune response to infection. In contrast to other viral chemokine-binding proteins, which interact promiscuously with multiple chemokines, pUL21.5 selectively binds RANTES (regulated upon activation, normal T cell expressed and secreted) with high affinity. By binding RANTES, pUL21.5 blocks RANTES interaction with its cellular receptors. We propose that human cytomegalovirus directs the synthesis of a secreted, virus-coded protein that modulates the host antiviral response even before the newly infecting viral genome becomes transcriptionally active. PMID- 15536130 TI - Greenhouse gas growth rates. AB - We posit that feasible reversal of the growth of atmospheric CH(4) and other trace gases would provide a vital contribution toward averting dangerous anthropogenic interference with global climate. Such trace gas reductions may allow stabilization of atmospheric CO(2) at an achievable level of anthropogenic CO(2) emissions, even if the added global warming constituting dangerous anthropogenic interference is as small as 1 degrees C. A 1 degrees C limit on global warming, with canonical climate sensitivity, requires peak CO(2) approximately 440 ppm if further non-CO(2) forcing is +0.5 W/m(2), but peak CO(2) approximately 520 ppm if further non-CO(2) forcing is -0.5 W/m(2). The practical result is that a decline of non-CO(2) forcings allows climate forcing to be stabilized with a significantly higher transient level of CO(2) emissions. Increased "natural" emissions of CO(2), N(2)O, and CH(4) are expected in response to global warming. These emissions, an indirect effect of all climate forcings, are small compared with human-made climate forcing and occur on a time scale of a few centuries, but they tend to aggravate the task of stabilizing atmospheric composition. PMID- 15536131 TI - The influence of large-scale wind power on global climate. AB - Large-scale use of wind power can alter local and global climate by extracting kinetic energy and altering turbulent transport in the atmospheric boundary layer. We report climate-model simulations that address the possible climatic impacts of wind power at regional to global scales by using two general circulation models and several parameterizations of the interaction of wind turbines with the boundary layer. We find that very large amounts of wind power can produce nonnegligible climatic change at continental scales. Although large scale effects are observed, wind power has a negligible effect on global-mean surface temperature, and it would deliver enormous global benefits by reducing emissions of CO(2) and air pollutants. Our results may enable a comparison between the climate impacts due to wind power and the reduction in climatic impacts achieved by the substitution of wind for fossil fuels. PMID- 15536132 TI - Dynamic histone modifications mark sex chromosome inactivation and reactivation during mammalian spermatogenesis. AB - Based on the formation of the XY body at pachytene and expression studies of a few X-linked genes, the X and Y chromosomes seem to undergo transcriptional inactivation during mammalian spermatogenesis. However, the extent and the mechanism of X and Y inactivation are not known. Here, we show that both the X and Y chromosomes undergo sequential changes in their histone modifications beginning at the pachytene stage of meiosis. These changes usually are associated with transcriptional inactivation in somatic cells, and they coincide with the exclusion of the phosphorylated (active) form of RNA polymerase II from the XY body. Both sex chromosomes undergo extensive deacetylation at histones H3 and H4 and (di)methylation of lysine (K)9 on histone H3; however, there are no changes in H3-K4 methylation. These changes persist even when the XY body disappears in late pachytene, and the X and Y chromosomes segregate from one another after the first meiotic division. By the spermatid stage, histone modifications of the X and Y chromosomes revert to those of active chromatin and RNA polymerase II reengages with both chromosomes. Our observations indicate that X and Y inactivation is extensive and persists even when the X and Y chromosomes are separated in secondary spermatocytes. These findings provide insights into epigenetic programming and chromatin dynamics in the male germ line. PMID- 15536133 TI - Lhx2-/- mice develop liver fibrosis. AB - Liver fibrosis is a wound-healing response to chronic injury of any type and is characterized by a progressive increase in deposition of extracellular matrix (ECM) proteins, the major source of which are activated hepatic stellate cells (HSCs). Because the LIM homeobox gene Lhx2 is expressed in HSCs and liver development in Lhx2(-/-) mice is disrupted, we analyzed liver development in Lhx2(-/-) embryos in detail. Lhx2(-/-) embryos contain numerous activated HSCs and display a progressively increased deposition of the ECM proteins associated with liver fibrosis, suggesting that Lhx2 inhibits HSC activation. Transfection of Lhx2 cDNA into a human HSC line down-regulates expression of genes characteristic of activated HSCs. Moreover, the Lhx2(-/-) liver display a disrupted cellular organization and an altered gene expression pattern of the intrahepatic endodermal cells, and the increased deposition of ECM proteins precedes these abnormalities. Collectively these results show that Lhx2 negatively regulates HSC activation, and its inactivation in developing HSCs appears therefore to mimic the signals that are triggered by the wound-healing response to chronic liver injury. This study establishes a spontaneous and reproducible animal model for hepatic fibrosis and reveals that Lhx2 expression in HSCs is important for proper cellular organization and differentiation of the liver. PMID- 15536134 TI - Recruitment of IkappaBalpha to the hes1 promoter is associated with transcriptional repression. AB - The NF-kappaB pathway plays a pivotal role in proliferation, differentiation, apoptosis, and immune responses in mammals. The NF-kappaB inhibitor, IkappaB, has classically been characterized for its ability to sequester NF-kappaB transcription factors in the cytoplasm. Nevertheless, a nuclear fraction of IkappaBalpha has consistently been detected and associated with repression of nuclear NF-kappaB. Now we show that IkappaBalpha physically associates with different repression elements such as nuclear corepressors and histone acetyltransferases and deacetylases (HDACs). More remarkably, chromatin immunoprecipitation experiments demonstrate that IkappaBalpha is recruited to the promoter regions of the Notch-target gene, hes1, together with HDAC1 and -5, whereas we did not detect IkappaBalpha associated with classical NF-kappaB target genes such as IL6 and RANTES. TNF-alpha treatment results in a temporary release of IkappaBalpha from the hes1 promoter that correlates with increased histone acetylation and transcriptional activation. In addition, we demonstrate that both IkappaB kinase-alpha and -beta are simultaneously recruited to the hes1 promoter in response to TNF-alpha, coinciding with a maximum of IkappaBalpha release and gene activation. Moreover, TNF-alpha-dependent histone H3 acetylation, release of IkappaBalpha from the hes1 promoter, and hes1 mRNA synthesis are affected in IKK alpha(-/-) mouse embryonic fibroblasts. We propose that IkappaBalpha plays a previously undescribed role in regulating the recruitment of repression elements to specific promoters. Recruitment of IKKs to the nucleus in response to TNF alpha may induce chromatin-associated IkappaBalpha release and gene activation. These findings provide additional insight in the cross-talk between NF-kappaB and other signaling pathways. PMID- 15536135 TI - Data mining, drug safety, and molecular pharmacology: potential for collaboration. PMID- 15536136 TI - Anaphylaxis to intravenous sinistrin. PMID- 15536137 TI - Updated clinical risk factor analysis of suspected hypersensitivity reactions to abacavir. PMID- 15536140 TI - Role of low-molecular-weight heparin in invasive management of non-ST-elevation acute coronary syndromes. AB - OBJECTIVE: To review the available literature addressing the role of low molecular-weight heparin (LMWH) as an alternative to unfractionated heparin (UFH) in percutaneous coronary intervention (PCI) for treatment of non-ST-elevation acute coronary syndromes (NSTEACS). DATA SOURCES: A MEDLINE search (1966-March 2004) identified pertinent articles using the key words acute coronary syndromes, unstable angina, non-ST-elevation myocardial infarction, low-molecular-weight heparin, enoxaparin, dalteparin, glycoprotein IIb/IIIa receptor antagonists, abciximab, tirofiban, eptifibatide, percutaneous transluminal coronary angioplasty, and percutaneous coronary intervention. The references of these articles were reviewed for additional pertinent references. STUDY SELECTION AND DATA EXTRACTION: All human trials of LMWH in PCI for treatment of NSTEACS were evaluated. All pertinent studies were included in the review. DATA SYNTHESIS: Administration of LMWH with or without a glycoprotein IIb/IIIa inhibitor during PCI appears to be similar to UFH in terms of efficacy. LMWH, especially in combination with a glycoprotein IIb/IIIa inhibitor, may increase risk of bleeding compared with UFH. CONCLUSIONS: Available clinical trials do not provide definitive evidence to suggest superiority of LMWH over UFH when managing NSTEACS during PCI; however, dosing strategies are available if an LMWH is to be used in this setting. PMID- 15536141 TI - Intraperitoneal cefazolin and ceftazidime effects on human peritoneal mesothelial cell release of cancer antigen-125. AB - BACKGROUND: Intraperitoneal (IP) cefazolin and ceftazidime are recommended as empiric treatment for peritoneal dialysis (PD)-associated peritonitis. Human peritoneal mesothelial cells (HPMCs) may be affected by high IP cefazolin and ceftazidime concentrations. Peritoneal dialysate cancer antigen-125 (CA-125) appearance rate can be used to measure HPMC damage. OBJECTIVE: To determine whether IP cefazolin and ceftazidime increase peritoneal CA-125 appearance rate. METHODS: The study consisted of 2 phases. In phase I, no antibiotic was administered, and in phase II, patients received IP cefazolin and ceftazidime (15 mg/kg rounded to nearest 100 mg). Phase II occurred immediately after phase I. Each phase used a 4-hour dwell time with 2 L of dextrose 2.5% dialysate. Dialysate samples were collected at 0, 0.5, 1, 2, and 4 hours during each phase. Samples were assayed for CA-125, and CA-125 appearance rate was calculated. RESULTS: Thirteen patients were recruited (7 men; aged 44.0 +/-16.0 y). The mean +/- SD (range) CA-125 dialysate concentration after phases I and II were 6.6 +/- 3.7 U/mL (2.3-15.0) and 6.4 +/-3.8 U/mL (1.6-13.8), respectively (p = 0.46). The CA-125 appearance rate after phases I and II were 51.9 +/- 31.3 U/min/1.73 m(2) (13.8-113.0) and 50.5 +/- 32.9 U/min/1.73 m(2) (11.0-104.0), respectively (p = 0.57). The slopes of the regression lines of CA-125 appearance rate were not significantly different between phases I and II. CONCLUSIONS: These findings demonstrate that concurrently administered IP cefazolin and ceftazidime have no effect on HPMC release of CA-125 in non-infected PD patients. PMID- 15536142 TI - Association of aspirin with eosinophilia in peripheral blood. PMID- 15536143 TI - Acute renal failure and intravenous immune globulin: occurs with sucrose stabilized, but not with D-sorbitol-stabilized, formulation. AB - OBJECTIVE: To report 2 cases of acute renal failure (ARF) following administration of sucrose-stabilized intravenous immune globulin (IVIG), one of which did not recur following subsequent doses of d-sorbitol-stabilized formulation, and review the relevant literature. CASE SUMMARIES: A 44-year-old white man awaiting heart transplantation developed ARF requiring hemodialysis following administration of sucrose-stabilized IVIG for high alloreactivity to population human leukocyte antigens. Following a return of renal function to baseline, subsequent doses of d-sorbitol-stabilized IVIG were administered without incident. A 90-year-old white man developed ARF after administration of sucrose-stabilized IVIG for monoclonal gammopathy. Renal function returned to baseline, and no subsequent IVIG doses were administered. An objective causality assessment revealed that sucrose-stabilized IVIG was the probable cause of the adverse drug event for both cases. DISCUSSION: Several case reports of ARF secondary to IVIG have been published. Recent publications note that sucrose stabilized IVIG products have a disproportionately high rate of ARF occurrence (approximately 88%) versus non-sucrose-stabilized formulations. Recent market data for IVIG products indicate that sucrose-stabilized products account for approximately 40% of the total IVIG market. When administered intravenously, sucrose is excreted unchanged in the urine. ARF has been reported in patients receiving large doses of intravenous sucrose. CONCLUSIONS: ARF secondary to IVIG may be more likely to occur with sucrose-stabilized formulations. Before prescribing IVIG, clinicians should consider other nephrotoxic medications, preexisting renal function, age, diabetes mellitus, and rate of infusion. In patients at risk, it may be best to avoid sucrose-stabilized formulations. PMID- 15536144 TI - Analysis of the receptor-ligand interactions in the natural killer-mediated lysis of freshly isolated myeloid or lymphoblastic leukemias: evidence for the involvement of the Poliovirus receptor (CD155) and Nectin-2 (CD112). AB - On the basis of recent clinical and experimental data, natural killer (NK) cells appear to play a crucial role in eradication of acute myeloid leukemias. In the present study, by exploiting our current knowledge on NK receptors and their ligands on target cells, we investigated the interactions between NK and leukemic cells. We show that the size of the NK cell subset expressing the killer immunoglobulin-like receptor (KIR) not engaged by the HLA-class I alleles of the patient parallels the degree of NK cytotoxicity against leukemic cells. A sharp down-regulation of HLA-class I molecules has been detected in various leukemias and it was more frequent in myeloid than in lymphoblastic leukemias. Analysis of the ligands for triggering NK receptors revealed the consistent expression of Poliovirus receptor (PVR) and Nectin-2 in myeloid leukemias. In contrast, major histocompatibility complex class I-related chain molecules A/B (MICA/B) and UL1b binding protein (ULBPs) were either absent or weakly expressed. Accordingly, NK mediated lysis of these leukemias was dependent on DNAM-1 but not NKG2D. The major role of NKp46 and NKp30 was also confirmed. The expression of PVR and/or Nectin-2 was less frequent in lymphoblastic leukemias. In most leukemias, both CD48 and NTBA were down-regulated. The correlation found between marker expression and susceptibility to lysis may reveal useful information for NK-based immunotherapy. PMID- 15536145 TI - NK cells that are activated by CXCL10 can kill dormant tumor cells that resist CTL-mediated lysis and can express B7-H1 that stimulates T cells. AB - Tumor dormancy is a phenomenon where small numbers of tumor cells persist in the host for months or years. We previously showed in the DA1-3b/C3H mouse model of acute myeloid leukemia that dormant tumor cells resist cytotoxic T-lymphocyte (CTL)-mediated killing because they overexpress B7-H1. Here, we vaccinated mice with DA1-3b cells transduced with CXCL10. Vaccinated mice developed a strong systemic immunity that led to the cure of established leukemia without persistence of dormant tumor cells. In vivo depletion of natural killer (NK) cells from the mice abrogated the protective effect of the vaccine. Long-term persistent leukemic cells resist CTL-mediated lysis but were killed by NK cells from mice vaccinated with DA1-3b/CXCL10. These NK cells expressed B7-H1. Recombinant CXCL10, CXCL9, CXCL11, and CXCL12 chemokines induced expression of B7 H1 on mouse and human NK cells in vitro. Mouse and human B7-H1+ NK cells induced proliferation of T cells and production of interferon gamma and tumor necrosis factor alpha in vitro, and in vivo blocking of B7-H1 inhibited the protective effect of vaccination. Thus, CXCL10 induces antileukemic immunity, at least partially by stimulating NK cells to express B7-H1+. This antitumor effect is in contrast to the effect of B7-H1 when expressed on tumor cells because it stops cytotoxic lymphocytes from killing those tumor cells. PMID- 15536146 TI - TRAIL identifies immature natural killer cells in newborn mice and adult mouse liver. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a key effector molecule expressed by natural killer (NK) cells and has been shown to prevent tumor initiation, growth, and metastasis. Here we demonstrate that TRAIL is the dominant cytotoxic effector molecule expressed by NK cells in fetal mice. On birth and with age, NK cells develop full functional capacity, including the ability to secrete interferon gamma (IFN-gamma) and interleukin 13 (IL-13) and mediate perforin- and Fas ligand-mediated cytotoxicity. However, interestingly, a phenotypically immature TRAIL+ NK cell subpopulation is retained in the liver of adult mice, and its retention is dependent on IFN-gamma but not dependent on host IL-12, IL-18, or endogenous host pathogens. Adoptive transfer of either adult liver or neonatal TRAIL+ NK cells resulted in the appearance of TRAIL- NK cells with a mature phenotype, suggesting that these TRAIL+ NK cells were indeed a precursor. Although inducers of IFN-gamma stimulated TRAIL expression on mature NK cells, our data indicated that constitutive TRAIL expression was a hallmark of immature cytotoxic NK cells. This study is the first to describe the concomitant maturation of NK cell effector function with surface phenotype in vivo and implies an important defense role for NK cell TRAIL in the developing immune system. PMID- 15536147 TI - Molecular transfer of CD40 and OX40 ligands to leukemic human B cells induces expansion of autologous tumor-reactive cytotoxic T lymphocytes. AB - Clinical benefits from monoclonal antibody therapy for B-chronic lymphocytic leukemia (B-CLL) have increased interest in developing additional immunotherapies for the disease. CD40 ligand is an accessory signal for T-cell activation and can overcome T-cell anergy. The OX40-OX40 ligand pathway is involved in the subsequent expansion of memory antigen-specific T cells. We expressed both CD40L and OX40L on B-CLL cells by exploiting the phenomenon of molecular transfer from fibroblasts overexpressing these ligands. We analyzed the effects of the modified B-CLL cells on the number, phenotype, and cytotoxic function of autologous T cells in 7 B-CLL patients. Transfer of CD40L and OX40L was observed in all and was followed by the up-regulation of B7-1 and B7-2. The culture of CD40L/OX40L expressing B-CLL cells with autologous T cells generated CD4+/CD8+ cytotoxic T cell lines, which secreted interferon-gamma (IFN-gamma) and granzyme-B/perforin in response to autologous, but not to allogeneic, B-CLL cells or to autologous T cell blasts. CD40L or OX40L alone was insufficient to expand tumor-reactive T cells. The combination of CD40L and OX40L on B-CLL cells may allow the generation of therapeutic immune responses to B-CLL, either by active immunization with modified tumor cells or by adoptive immunotherapy with tumor-reactive autologous T cells. PMID- 15536148 TI - Genotypic inhibitory killer immunoglobulin-like receptor ligand incompatibility enhances the long-term antileukemic effect of unmodified allogeneic hematopoietic stem cell transplantation in patients with myeloid leukemias. AB - It remains controversial whether alloreactive donor-derived natural killer (NK) cells display graft-versus-leukemia reactions after unmodified allogeneic hematopoietic stem cell transplantation (HSCT). The present study evaluated the role of inhibitory killer immunoglobulin-like receptor (KIR) ligand incompatibility using a well-defined and uniform setting of unmodified allogeneic HSCT in 374 patients with myeloid leukemias. The most striking finding was a significant heterogeneity in the 5-year estimates of hematologic leukemic relapse after human leukocyte antigen (HLA)-identical (n = 237; 22%), HLA class I disparate (n = 89; 18%), and KIR ligand-incompatible transplantations (n = 48; 5%) (P < .04). Multivariate analysis confirmed that the relative relapse risk (RR) was influenced by HLA class I disparity alone (RR 0.49), but was lowest after HLA class I-disparate, KIR ligand-incompatible transplantations (RR 0.24) (P < .008). The primary graft failure rates, however, increased from 0.4% after HLA class I-identical to 2.3% after HLA class I-disparate, and to 6.3% after KIR ligand-incompatible transplantations, respectively (P < .02). Unlike some other reports, no beneficial effect of KIR ligand incompatibility on other major endpoints of allogeneic HSCT (transplantation-related mortality, and overall and event-free survival) was detectable in the present study. In conclusion, unmodified allogeneic HSCT from KIR ligand-incompatible donors provides a superior long-term antileukemic efficacy in patients with myeloid malignancies. PMID- 15536149 TI - RGS2 is an important target gene of Flt3-ITD mutations in AML and functions in myeloid differentiation and leukemic transformation. AB - Activating fetal liver tyrosine kinase 3 (Flt3) mutations represent the most common genetic aberrations in acute myeloid leukemia (AML). Most commonly, they occur as internal tandem duplications in the juxtamembrane domain (Flt3-ITD) that transform myeloid cells in vitro and in vivo and that induce aberrant signaling and biologic functions. We identified RGS2, a regulator of G-protein signaling, as a gene specifically repressed by Flt3-ITD. Here we demonstrate an important role of RGS2 in Flt3-ITD-mediated transformation. RGS2 was repressed after forced expression of activating Flt3 mutations in 2 myeloid cell lines (32Dcl3 and NB4). Furthermore, RGS2 was repressed in Flt3-mutation-positive AML cases in comparison to Flt3-mutation-negative cases, especially in Flt3-ITD-positive cases with a high ITD-to-wild-type (WT) ratio. Coexpression of RGS2 with Flt3-ITD inhibited Flt3-ITD-induced autonomous proliferation and clonal growth of 32D cells. RGS2 also inhibited Flt3-ITD-induced phosphorylation of Akt and glycogen synthase kinase beta (Gsk3-beta) without influencing signal transducer and activator of transcription 5 (STAT5) activation. In addition, RGS2 reinduced the expression of Flt3-ITD-repressed CCAAT/enhancer-binding protein alpha (c/EBPalpha) and antagonized the Flt3-ITD-induced differentiation block in 32D cells. Expression analyses in myeloid cell lines revealed induction of RGS2 during granulocytic but not during monocytic differentiation. Taken together, RGS2 is a novel mediator of myeloid differentiation, and its repression is an important event in Flt3-ITD induced transformation. PMID- 15536150 TI - Development and validation of a clinical prediction rule for bone marrow involvement in patients with Hodgkin lymphoma. AB - We developed a clinical prediction rule for bone marrow involvement (BMI) in Hodgkin lymphoma based on 826 patients and validated it in 654 additional patients. Independent prognostic factors for BMI were x1, B symptoms; x2, stage III/IV prior to bone marrow biopsy; x3, anemia; x4, leukocytes fewer than 6 x 10(9)/L; x5, age 35 years or older; and x6, iliac/inguinal involvement. Each factor was graded as x(i)=1, if present, or x(i)=0, if absent. A simplified score Zs=8x1+6x2+5x3+5x4+3x5+3x6-8 was assigned to each patient. The sensitivity, specificity, and positive and negative predictive value of this prediction rule was 97.8%, 51.5%, 10.6%, and 99.8%, respectively. In the validation group, they were 98.1%, 40.3%, 12.7%, and 99.6%. According to Zs value, 3 risk groups for BMI were defined: low risk (Zs<0, 44% of patients, 0.3% risk), standard risk (Zs, 0 9; 37% of patients; 4.2% risk), and high risk (Zs>or=10, 20% of patients, 25.5% risk). Patients with low risk (stage IA/IIA without anemia and leukopenia; stage IA/IIA, younger than 35 years, with either anemia or leukopenia but no inguinal/iliac involvement; and stage IIIA/IVA without any of these 4 risk factors) do not need bone marrow (BM) biopsy. Patients with standard risk should be staged with unilateral biopsy, but patients with high risk may benefit from bilateral biopsy. PMID- 15536151 TI - The Corfu deltabeta thalassemia deletion disrupts gamma-globin gene silencing and reveals post-transcriptional regulation of HbF expression. AB - The 7.2 kilobase (kb) Corfu deltabeta thalassemia mutation is the smallest known deletion encompassing a region upstream of the human delta gene that has been suggested to account for the vastly different phenotypes in hereditary persistence of fetal hemoglobin (HPFH) versus beta thalassemia. Fetal hemoglobin (HbF) expression in Corfu heterozygotes and homozygotes is paradoxically dissimilar, suggesting conflicting theories as to the function of the region on globin gene regulation. Here, we measure gamma- and beta-globin gene transcription, steady-state mRNA, and hemoglobin expression levels in primary erythroid cells cultured from several patients with Corfu deltabeta thalassemia. We show through RNA fluorescence in situ hybridization that the Corfu deletion results in high-level transcription of the fetal gamma genes in cis with a concomitant reduction in transcription of the downstream beta gene. Surprisingly, we find that elevated gamma gene transcription does not always result in a corresponding accumulation of gamma mRNA or fetal hemoglobin, indicating a post transcriptional regulation of gamma gene expression. The data suggest that efficient gamma mRNA accumulation and HbF expression are blocked until beta mRNA levels fall below a critical threshold. These results explain the Corfu paradox and show that the deleted region harbors a critical element that functions in the developmentally regulated transcription of the beta-globin genes. PMID- 15536152 TI - Lymphatic dysfunction in transgenic mice expressing KSHV k-cyclin under the control of the VEGFR-3 promoter. AB - Kaposi sarcoma-associated herpesvirus (KSHV) infects endothelial cells within KS tumors, and these cells express the KSHV latent-cycle gene k-cyclin (kCYC) as well as vascular endothelial growth factor receptor 3 (VEGFR-3), a marker for lymphatic endothelium. To further understand KSHV-mediated pathogenesis, we generated transgenic mice expressing kCYC under the control of the VEGFR-3 promoter. kCYC mRNA and functional protein expression within tissue correlated with VEGFR-3 expression and were most abundantly detected within lung tissue. Clinically, most transgenic mice died within 6 months of age secondary to progressive accumulation of chylous pleural fluid. In skin, edema was detected by magnetic resonance imaging and mice demonstrated persistent erythema of the ears following trauma. Histologically, erythematous skin showed extravasation of erythrocytes and accumulation of erythrocytes within lymphatic lumens. In addition, lymphatic drainage of injected contrast dyes was markedly impaired in transgenic mice. Karyomegaly, a feature observed in kCYC-expressing cells in vitro, was detected in many tissues, and selectively occurred within lymphatic endothelial cells expressing kCYC mRNA by in situ hybridization. In summary, kCYC expression within VEGFR-3+ cells of mice causes marked impairment of lymphatic function. kCYC may contribute to the development of certain clinical and histologic features of KS, including localized edema and retention of extravasated erythrocytes within KS tumors. PMID- 15536153 TI - WHIM syndromes with different genetic anomalies are accounted for by impaired CXCR4 desensitization to CXCL12. AB - The WHIM syndrome is a rare immunodeficiency disorder characterized by warts, hypogammaglobulinemia, infections, and myelokathexis. Dominant heterozygous mutations of the gene encoding CXCR4, a G-protein-coupled receptor with a unique ligand, CXCL12, have been associated with this pathology. We studied patients belonging to 3 different pedigrees. Two siblings inherited a CXCR4 mutation encoding a novel C-terminally truncated receptor. Two unrelated patients were found to bear a wild-type CXCR4 open reading frame. Circulating lymphocytes and neutrophils from all patients displayed similar functional alterations of CXCR4 mediated responses featured by a marked enhancement of G-protein-dependent responses. This phenomenon relies on the refractoriness of CXCR4 to be both desensitized and internalized in response to CXCL12. Therefore, the aberrant dysfunction of the CXCR4-mediated signaling constitutes a common biologic trait of WHIM syndromes with different causative genetic anomalies. Responses to other chemokines, namely CCL4, CCL5, and CCL21, were preserved, suggesting that, in clinical forms associated with a wild-type CXCR4 open reading frame, the genetic anomaly might target an effector with some degree of selectivity for the CXCL12/CXCR4 axis. We propose that the sustained CXCR4 activity in patient cells accounts for the immune-hematologic clinical manifestations and the profusion of warts characteristic of the WHIM syndrome. PMID- 15536154 TI - Gonadotropin-releasing hormone antagonist cetrorelix down-regulates proliferating cell nuclear antigen and epidermal growth factor expression and up-regulates apoptosis in association with enhanced poly(adenosine 5'-diphosphate-ribose) polymerase expression in cultured human leiomyoma cells. AB - The objective of this study was to elucidate the effects of GnRH antagonist Cetrorelix on proliferation and apoptosis in human leiomyoma cells cultured in vitro. Isolated leiomyoma cells were subcultured in phenol red-free DMEM supplemented with 10% fetal bovine serum for 120 h and then stepped down to serum free conditions in the presence or absence of graded concentrations of Cetrorelix (10(-5) to 10(-8) mol/liter) for 6 d. Cultured leiomyoma cells were used for semiquantitative RT-PCR, immunocytochemistry, Western blot analysis, and terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling assay. RT-PCR analysis revealed the presence of mRNAs encoding for GnRH receptor and epidermal growth factor (EGF) in cultured leiomyoma cells. The number of viable cultured leiomyoma cells was significantly (P < 0.01) decreased by treatment with Cetrorelix compared with untreated control cultures. Immunocytochemical examination demonstrated that treatment with Cetrorelix attenuated the expression of proliferating cell nuclear antigen (PCNA) and EGF in cultured leiomyoma cells. Western blot analysis revealed that treatment with 10( 5) mol/liter Cetrorelix significantly (P < 0.01) decreased PCNA expression. In addition, treatment with 10(-5) mol/liter Cetrorelix remarkably increased the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick end labeling-positive rate and poly(ADP-ribose) polymerase expression at 24 h of treatment compared with untreated control cultures (P < 0.01). Furthermore, treatment with 10(-5) mol/liter Cetrorelix decreased immunoreactive EGF protein and EGF mRNA expression in cultured leiomyoma cells at 4 d of treatment. GnRH antagonist Cetrorelix may directly inhibit leiomyoma cell growth by down regulating proliferation in association with a decrease in EGF mRNA expression and by up-regulating apoptosis in those cells. PMID- 15536155 TI - Regulation of matrix metalloproteinase-9/gelatinase B expression and activation by ovarian steroids and LEFTY-A/endometrial bleeding-associated factor in the human endometrium. AB - Various matrix metalloproteinases (MMPs) participate in the menstrual breakdown of the human endometrium. MMP-9/gelatinase B is proposed as a major factor because it degrades many extracellular matrix constituents, including in the vasculature. Although globally under ovarian steroids control, endometrial MMP-9 seems expressed differently than other MMPs, and conflicting publications prevent a clear understanding of its regulation. We therefore quantified MMP-9 expression in the cycling human endometrium, defined its localization, and analyzed its regulation by estradiol and progesterone and by LEFTY-A/endometrial bleeding associated factor in explant cultures. In fresh tissues, a major increase in MMP 9 mRNA expression occurred at menstruation, after a larger increase in LEFTY-A mRNA. MMP-9 was immunodetected in all cell types throughout the cycle, especially in foci of stromal cells during menstruation. MMP-9 synthesis by these cells was confirmed in cultured explants. In proliferative explants, ovarian steroids slightly decreased MMP-9 mRNA. They had no consistent effect on MMP-9 release in culture medium but strongly inhibited proMMP-9 activation. Addition of recombinant LEFTY-A to explants induced MMP-9 in most samples, a response prevented by ovarian steroids. We propose that endometrial MMP-9 activity is overall controlled by the ovarian steroids and locally adjusted through a network of modulators, including LEFTY-A. PMID- 15536156 TI - Correlating androgen and estrogen steroid receptor expression with coronary calcification and atherosclerosis in men without known coronary artery disease. AB - BACKGROUND: Accumulating data emphasize the gender specificity of key components of the atherosclerotic process and the importance of gonadal steroids on the human vasculature. Steroid receptors, including the androgen receptor (AR) and estrogen receptors (ERs) alpha and beta are expressed in key vascular tissues, including endothelial cells and vascular smooth muscle cells. However, the relative abundance and importance of these receptors in the coronary artery are not well defined, particularly in men. We therefore examined AR, ER alpha, and ER beta expression as a function of key components of atherosclerosis, namely plaque and calcium area, in male human coronary arteries. METHODS: Coronary arteries were obtained at autopsy from 24 men without known coronary artery disease. Coronary calcification was measured by contact microradiography, and atherosclerotic plaque area was quantified histologically. Coronary artery cross sections were immunostained for AR, ER alpha, and ER beta and then measured semiquantitatively in each arterial wall layer (intima, adventitia, and media). RESULTS: AR, ER beta, and ER alpha were expressed in all artery wall layers but most avidly in the media (P < 0.001). ER beta exceeded ER alpha expression (P < 0.0005). AR expression in the media correlated negatively with plaque area (P = 0.006, R = -0.55), whereas intimal ER beta expression correlated positively with plaque area (P = 0.012, R = 0.50). CONCLUSIONS: We conclude that both AR and ER beta are important in relatively early coronary atherosclerosis, but inversely so, because decreasing AR and increasing ER beta expression correlate with more extensive atherosclerosis. ER beta seems to be the predominate ER in coronary arteries harvested from men without known coronary artery disease. Interventional studies are required to assess the functional significance of these observations. PMID- 15536157 TI - Ghrelin stimulates, whereas des-octanoyl ghrelin inhibits, glucose output by primary hepatocytes. AB - Ghrelin exerts various metabolic activities, including regulation of glucose levels in humans. To verify whether the glucose response to ghrelin reflects a modulation of an insulin-independent hepatic phenomenon, we studied glucose output by primary porcine hepatocytes in suspension culture, after incubation with acylated ghrelin (AG), unacylated ghrelin (UAG), and hexarelin (HEX). AG induced glucose output dose dependently after 20 min of incubation (P < 0.001), whereas HEX, a GH secretagogue receptor type 1a (GHS-R1a) agonist, had no effect. UAG inhibited glucose release also dose dependently and after 20 min (P < 0.001). Moreover, UAG completely reversed AG-induced glucose output (P < 0.01). Using real-time PCR, GHS-R1a gene expression was undetectable in all the hepatocyte preparations studied. The lack of efficacy of HEX, the efficacy of UAG, and the absence of GHS-R1a expression indicate the involvement of a yet uncharacterized ghrelin receptor type. In conclusion, glucose output by primary hepatocytes is time- and dose-dependently stimulated by AG and inhibited by UAG. Moreover, UAG counteracts the stimulatory effect of AG on glucose release. These actions might be mediated by a different receptor than GHS-R1a, and apparently, we must consider AG and UAG as separate hormones that can modify each other's actions on glucose handling, at least in the liver. PMID- 15536158 TI - The metabolic syndrome and smoking in relation to hypogonadism in middle-aged men: a prospective cohort study. AB - In men, hypoandrogenism is associated with features of the metabolic syndrome. It is not known whether men with the metabolic syndrome are at a higher risk of developing hypogonadism. We therefore assessed whether the metabolic syndrome predicts development of hypogonadism 11 yr later in 651 middle-aged Finnish men participating in a population-based cohort study. Men with the metabolic syndrome at baseline as defined by the World Health Organization (n = 114, 20%) had a 2.6 fold increased risk of developing hypogonadism as defined by total testosterone levels less than 11 nmol/liter at the 11-yr follow-up independent of age, smoking, and other potential confounders. Further adjustment for body mass index (OR, 2.0; 95% CI, 1.1-3.8) or baseline total testosterone levels (OR, 1.9; 95% CI, 1.0-3.4) attenuated the association. The association of the metabolic syndrome with hypogonadism as defined by calculated free testosterone levels less than 225 pmol/liter was similar, but weaker. The adjusted decrease in testosterone concentrations during the 11-yr follow-up was also greater in men with than without the metabolic syndrome. Smokers had a nonsignificantly lower risk of developing hypogonadism during follow-up, whereas a decrease in smoking increased the risk of hypogonadism. The metabolic syndrome predisposes to development of hypogonadism in middle-aged men. Prevention of abdominal obesity and the accompanying metabolic syndrome in middle age may decrease the risk of hypogonadism in men, especially in those who quit smoking. PMID- 15536159 TI - Detection of monoclonality of the immunoglobulin heavy chain gene in thyroid malignant lymphoma by vectorette polymerase chain reaction. AB - Distinguishing between thyroid malignant lymphoma and lymphocytic thyroiditis (Hashimoto's thyroiditis) is quite difficult and problematic. B cell lymphomas display clonal Ig heavy-chain (IgH) gene rearrangement, and Southern blot hybridization (SBH) is often used for detection of the monoclonality of the IgH gene. However, SBH is often problematic because it requires a large volume of samples. We examined the efficiency in detecting the monoclonality of IgH gene in thyroid malignant lymphomas by vectorette PCR, which we started with only 200 ng of genomic DNA. Monoclonality was detected in 36 of 47 (76.6%) malignant lymphomas, whereas it was not detected in 10 samples of tissue of Hashimoto's thyroiditis. The sensitivity was almost the same as that with SBH in which monoclonality was detected in 33 of 45 (73.3%) malignant lymphomas. These results suggest that vectorette PCR may be a substitute for SBH, and because it requires only a small volume of samples, it may be used in the analysis of aspiration biopsies. PMID- 15536160 TI - Decreased muscle capillary permeability surface area in type 2 diabetic subjects. AB - Capillary recruitment in muscles, induced by insulin, has been proposed to be impaired in insulin-resistant states. To elucidate the mechanisms regulating capillary transport of insulin and glucose in type 2 diabetes, we directly calculated the permeability-surface area product (PS) for glucose and insulin in muscle. Intramuscular microdialysis in combination with the forearm model and blood flow measurements was performed in type 2 diabetic male subjects and age- and weight-matched controls during a euglycemic-hyperinsulinemic clamp. During steady-state hyperinsulinemia, arterial plasma glucose was 5.8 +/- 0.1 and 5.9 +/ 0.1 mmol/liter [not significant (NS)] in the obese and type 2 diabetic subjects, respectively. Venous glucose was significantly lower in the obese group compared with the type 2 diabetic subjects, 4.3 +/- 02 vs. 4.9 +/- 0.2 mmol/liter (P < 0.05). Arterial insulin was 1494 +/- 90 and 1458 +/- 132 pmol/liter (NS) in the obese and type 2 diabetic subjects, respectively. The glucose infusion rate during steady-state hyperinsulinemia was 10.8 +/- 0.8 and 7.2 +/- 0.4 mg/kg.min in the obese and diabetic subjects, respectively (P < 0.01). Interstitial arterial lactate difference was significantly higher in the obese subjects. During steady-state hyperinsulinemia, PS for glucose was significantly higher in the obese subjects (1.1 +/- 0.2 vs. 0.5 +/- 0.1 ml/min.100 g, P < 0.05). Glucose uptake was also significantly higher in the obese subjects (3.0 +/- 0.4 vs. 1.8 +/- 0.3 mumol/min.100 g, P < 0.05). During steady-state hyperinsulinemia, PS for insulin was 0.4 +/- 0.1 and 0.3 +/- 0.1 ml/min.100 g in the obese and diabetic subjects, respectively (NS), and insulin uptake was 258 +/- 54 vs. 168 +/- 24, respectively (NS). When both subject groups were pooled together, a significant correlation was found between PS for glucose and glucose uptake during steady state hyperinsulinemia. Skeletal muscle blood flow during steady-state hyperinsulinemia was 1.9 +/- 0.2 and 2.3 +/- 0.4 ml/100 g.min in the obese and diabetic subjects, respectively (NS). Blood flow did not increase during hyperinsulinemia in either of the two groups. The present data clearly show that PS for glucose is subnormal during steady-state hyperinsulinemia in insulin resistant type 2 diabetic subjects. Furthermore, there was a close correlation between glucose uptake and PS for glucose but not between blood flow and PS. We suggest that PS is a more sensitive marker for insulin resistance during hyperinsulinemia than limb flow. The lower capacity for transcapillary passage found in the type 2 diabetic subjects is suggested to further aggravate insulin resistance. PMID- 15536161 TI - Testicular anti-mullerian hormone secretion is stimulated by recombinant human FSH in patients with congenital hypogonadotropic hypogonadism. AB - Serum anti-Mullerian hormone (AMH), a prepubertal Sertoli cell marker, declines during puberty as an early sign of testicular testosterone (T) production. When T synthesis or action is impaired, serum AMH is abnormally high in the first months after birth and at puberty but normal between these two periods. We postulated that FSH might be responsible for AMH up-regulation in the absence of androgen inhibition. To test this hypothesis, we administered recombinant human (rh) FSH to eight patients aged from 18-31 yr with untreated congenital hypogonadotropic hypogonadism. This situation is ideal to study the effect of FSH on AMH production because it avoids interference by endogenous gonadotropins and T. The patients received daily sc injections of 150 IU rhFSH for 1 month, followed in seven of them by a combined treatment of rhFSH plus human chorionic gonadotropin (hCG; 1500 UI im, twice a week) for 2 months. Gonadotropins, T, AMH, and inhibin B were measured in plasma before treatment every 10 d during rhFSH treatment and every month during combined rhFSH and hCG treatments. All hormones were at prepubertal levels before treatment. Although LH and T did not vary, AMH and inhibin B levels gradually increased after 20 d of FSH administration. However, in contrast to rhFSH alone, the combined rhFSH plus hCG stimulation of the testis dramatically suppresses the secretion of AMH and induced a modest but significant reduction of circulating inhibin B levels. We conclude that FSH stimulates AMH production in the testis when it is at a prepubertal stage. In addition, the decrease of serum AMH during combined rhFSH and hCG testicular stimulation is in agreement with the concept that during pubertal development and in adult life, the suppressive effect of LH-driven testicular androgens outweighs the stimulating effect of FSH on AMH production by Sertoli cells. Finally, the hCG induced decrease in inhibin B suggests that in humans, as previously demonstrated in monkeys, testicular T is also able to inhibit inhibin B secretion. PMID- 15536162 TI - Orlistat is as beneficial as metformin in the treatment of polycystic ovarian syndrome. AB - The objective of this study was to evaluate and compare the effect of treatment with orlistat vs. metformin on the hormonal and biochemical features of patients with polycystic ovarian syndrome (PCOS). Twenty-one Caucasian women with PCOS [mean (+/-SEM) age 27 +/- 0.9 yr and body mass index 36.7 +/- 3.3 kg/m(2)] participated in this prospective, randomized, open-labeled study. All subjects had an 8-wk run-in period of dietary modification and then randomized to receive either metformin (500 mg three times daily) or orlistat (120 mg three times daily) for 3 months. Weight, blood pressure, and fasting blood samples were taken at screening, randomization, and on completion. Insulin resistance (IR) was calculated using the homeostasis model of assessment (HOMA)-IR method [HOMA-IR = (insulin x glucose)/22.5]. The results are expressed as mean +/- SEM. When compared with baseline, treatment with both orlistat [93.5 +/- 11.5 ng/dl (3.24 +/- 0.4 nmol/liter) vs. 114.5 +/- 11.5 ng/dl (3.97 +/- 0.4 nmol/liter), P = 0.039] and metformin [97.2 +/- 11.5 ng/dl (3.37 +/- 0.4 nmol/liter) vs. 120.0 +/- 8.7 ng/dl (4.16 +/- 0.3 nmol/liter), P = 0.048] produced a significant reduction in total testosterone. Treatment with orlistat produced a 4.69% reduction in weight (99.0 +/- 6.0 vs. 94.6 +/- 6.1 kg, P = 0.002), and this reduction was more significant than the reduction produced by metformin (4.69 vs. 1.02%, P = 0.006). There was no significant reduction seen after either treatment group for fasting insulin, HOMA-IR, SHBG, or any of the lipid parameters studied. In this study, orlistat produced a significant reduction in weight and total testosterone. The reduction in total testosterone was similar to that seen after treatment with metformin. Therefore, orlistat may prove to be a useful adjunct in the treatment of PCOS. PMID- 15536163 TI - A novel C-terminal growth hormone receptor (GHR) mutation results in impaired GHR STAT5 but normal STAT-3 signaling. AB - GH insensitivity (GHI) is an autosomal recessive disorder caused by defects in the GH receptor (GHR). In a 17-yr-old female with severe short stature and biochemical features of GHI, sequencing of GHR gene revealed a compound heterozygosity for two novel mutations: C83X and a G deletion at position 1776 (1776del). 1776del is predicted to result in GHR truncation to 581 amino acids with a nonsense sequence of residues 560-581. To clarify the effect of 1776del on GHR function, wild-type GHR, GHR-1776del, and two additional GHR mutants, GHR L561X (stop codon at site of the 1776del) and GHR-I582X (translation termination in GHR-1776del) were transiently expressed in CHO cells. After incubation with recombinant human GH, GHR-1776del showed lower signal transducer and activator of transcription 5 (STAT5)-mediated transcriptional activation ( approximately 50%, P < 0.05), as well as STAT5 Tyr694 phosphorylation (P < 0.05) compared with wild type GHR, whereas GHR-L561X and GHR-I582X showed normal STAT5 phosphorylation and transcriptional activity. In contrast, all vectors produced similar effects on STAT3-mediated transcriptional activation. In conclusion, this novel GHR-1776del mutation in a classical GHI patient illustrates an important mechanism of impaired GHR-STAT5 but intact GHR-STAT3 signaling. This effect might result from interference of C-terminal nonsense sequence in mutated GHR with STAT5 docking to upstream tyrosine residues. PMID- 15536164 TI - Up-regulation of the extracellular matrix remodeling genes, biglycan, neutrophil gelatinase-associated lipocalin, and matrix metalloproteinase-9 in familial amyloid polyneuropathy. AB - Familial amyloid polyneuropathy (FAP) is characterized by extracellular deposition of transthyretin (TTR) aggregates and amyloid fibrils, particularly in the peripheral nervous system (PNS) and is accompanied with changes in connective tissue. Given the invasiveness of nerve biopsy, FAP salivary glands (SGs) were used in microarray analysis; biglycan and neutrophil gelatinase-associated lipocalin (NGAL), two genes related to extracellular matrix (ECM) remodeling were overexpressed in FAP. Results were validated by RT-PCR and immunohistochemistry both in SG and in nerve biopsies of different stages of disease progression. Matrix metalloproteinase-9 (MMP-9), which exists as a complex with NGAL, was also increased in FAP and in vitro degraded TTR aggregates and fibrils; however in the presence of serum amyloid P, a universal amyloid component, TTR fibrils became resistant to MMP-9 proteolysis. Biglycan, NGAL, and MMP-9 are transcriptionally up-regulated by NF-kappaB, a transcription factor that is activated in FAP nerves and SG. Given the relationship between inflammation and ECM remodeling, and the increase of proinflammatory cytokines in FAP, IL-10 expression in FAP nerves was investigated; IL-10 increased after fibril deposition, suggesting a balance between proinflammatory and anti-inflammatory mechanisms. Changes in ECM-related proteins and inflammatory events may be relevant for therapy in FAP and other neurodegenerative disorders. PMID- 15536165 TI - Transforming growth factor-beta in acute renal failure: receptor expression, effects on proliferation, cellularity, and vascularization after recovery from injury. AB - Transforming growth factor (TGF)-beta1 and a number of TGF-beta-responsive genes are transiently enhanced following induction of ischemic acute renal failure (ARF) in the rat. The mRNA and protein expression of TGF-beta receptors were analyzed in postischemic rat kidneys by ribonuclease protection, in situ hybridization, and immunohistochemistry. TGF-betaRI and -RII were enhanced within 3 days of ischemia-reperfusion (I/R) injury and remained elevated for up 7 days post-I/R; TGF-beta receptor expression was localized primarily in regenerating tubules within the outer medulla. A neutralizing TGF-beta antibody exacerbated cellular proliferation observed on day 3 postischemia but had no effect on day 1 or 2. TGF-beta antibody treatment had no measurable effect on loss of renal function or the restoration of renal function during the recovery response for up to 35 days postsurgery. However, ischemic injury resulted in modest renal hypertrophy that is due, in part, to in an increase in the number of interstitial cells in the postischemic kidney. Immunohistochemistry showed that several of these cells stained positively for the fibroblast-specific marker, S100A4 positive. Anti-TGF-beta treatment substantially attenuated the renal hypertrophy, interstitial cellularity, and S100A4-positive cells present at 35 days post-I/R. Finally, TGF-beta immunoneutralization attenuated the loss of renal vascular density following recovery from I/R injury. These data suggest that the TGF beta/TbetaR system is enhanced in the postischemic kidney. However, the current study failed to identify a prominent role for this system in the repair of proximal tubules following ARF. In contrast, the activation of this system may play an important role in the long-term structure of the postischemic kidney by influencing microvascular structure and interstitial cellularity. PMID- 15536166 TI - The hyperbilirubinemic Gunn rat is resistant to the pressor effects of angiotensin II. AB - ANG II induces vasoconstriction, at least in part, by stimulating NADPH oxidase and generating reactive oxygen species. ANG II also induces heme oxygenase activity, and bilirubin, a product of such activity, possesses antioxidant properties. We hypothesized that bilirubin, because of its antioxidant properties, may reduce the pressor and prooxidant effects of ANG II. Our in vivo studies used the hyperbilirubinemic Gunn rat which is deficient in the enzyme uridine diphosphate glucuronosyl transferase, the latter enabling the excretion of bilirubin into bile. ANG II (0.5 mg x kg(-1) x day(-1)) or saline vehicle was administered by osmotic minipump to control and Gunn rats for 4 wk. The rise in systolic blood pressure induced by ANG II, as observed in control rats, was markedly reduced in Gunn rats, the latter approximately 50% less at 3 and 4 wk after the initiation of ANG II infusion. The chronic administration of ANG II also impaired endothelium-dependent relaxation responses in control rats but not in Gunn rats. As assessed by the tetrahydrobiopterin/dihydrobiopterin ratio, ANG II induced oxidative stress in the aorta in control rats but not in Gunn rats. Heightened generation of superoxide anion in aortic rings in ANG II-infused rats and by vascular smooth muscle cells exposed to ANG II was normalized by bilirubin in vitro. We conclude that the pressor and prooxidant effects of ANG II are attenuated in the hyperbilirubinemic Gunn rat, an effect which, we speculate, may reflect, at least in part, the scavenging of superoxide anion by bilirubin. PMID- 15536167 TI - Gene regulation of ENaC subunits by serum- and glucocorticoid-inducible kinase-1. AB - Aldosterone is a key regulator of epithelial Na+ channels (ENaC) in renal cortical collecting ducts (CCD). The goal of this study was to examine whether serum- and glucocorticoid-inducible kinase-1 (SGK1), an aldosterone-induced gene, is vital to the delayed effect of aldosterone by increasing the gene expression of ENaC subunits. To test this hypothesis, we compared the levels of ENaC mRNA in mouse CCD cells that stably express either full-length (FL)-SGK1 or a kinase-dead dominant negative (K127M)-SGK1. Our results revealed that SGK1 regulates gene expression of ENaC, whether cells are maintained in steroid-free media or in the presence of corticosteroids (CS) and/or other growth factors. Under all conditions, the loss of function of SGK1 caused a significant decrease in the expression of alpha- and beta-ENaC, but not gamma-ENaC. Compared with cells expressing FL-SGK1, K127M-SGK1 decreased the expression of alpha- and beta subunit mRNA by approximately 45 and approximately 90%, respectively. Next, to determine whether SGK1 is one of the proteins mediating the induction of alpha ENaC mRNA by CS, we compared steroid induction of alpha-ENaC in cells expressing K127M-SGK1 vs. FL-SGK1. The maximum level of alpha-ENaC mRNA levels following CS was significantly (approximately 45%) higher in FL-SGK1- vs. K127M-SGK1 expressing cells, although the fold-induction by CS was similar in both FL-SGK1- and K127M-SGK1-expressing cells. In summary, we report for the first time that SGK1 regulates transcription of ENaC subunits. We propose that the effect of SGK1 on ENaC transcription is mediated by the activation of unidentified transcription factors. PMID- 15536168 TI - Glomerular and tubular damage in normotensive and hypertensive rats. AB - Tubular cell damage is an important mediator of interstitial fibrosis in chronic renal diseases. Glomerular and tubular damage in genetic hypertension was therefore studied. Tubular and glomerular damage was investigated in 10-, 40-, and 70-wk-old spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) and compared with glomerular capillary pressure (P(GC)) and glomerulosclerosis in superficial (OC) and juxtamedullary (JMC). Tubular vimentin was used as criterion of tubular damage. Variation in tubular diameter was measured during change in perfusion pressure, and ureter ligation was used to demonstrate the relationship between tubular pressure and appearance of vimentin-positive cells. Tubular and glomerular damage was most pronounced in JMC and greater in SHR than in WKY. It was absent in 10-wk-old WKY and significantly higher in JMC of SHR compared with WKY at 70 wk of age. Numbers of vimentin-positive segments were 18 +/- 9 vs. 38 +/- 7% in JMC of 70-wk-old WKY and SHR (P < 0.02), and glomerulosclerosis was seen in 8 +/- 3 vs. 19 +/- 5% of glomeruli in JMC of 70-wk-old WKY and SHR, respectively (P < 0.01). P(GC) was 45 +/- 3 mmHg in JMC of WKY and 57 +/- 3 mmHg in JMC of 70-wk-old SHR (P < 0.001). Tubular diameter variation was greatest in SHR (P < 0.05) during pressure variation. Proteinuria was present only in 40- and 70-wk-old SHR and did not correlate with tissue damage. Tubular and glomerular damage in both strains develops in parallel and may be caused by a common mechanism, which may be glomerular capillary and tubular wall stretch during acute blood pressure variation which is greatest in JMC in SHR. PMID- 15536169 TI - Regulation of the energy sensor AMP-activated protein kinase in the kidney by dietary salt intake and osmolality. AB - The AMP-activated protein kinase (AMPK) is a key controller of cellular energy metabolism. We studied its expression and regulation by salt handling in the kidney. Immunoprecipitation and Western blots of protein lysates from whole rat kidney using subunit-specific antibodies showed that the alpha1-catalytic subunit is expressed in the kidney, associated with the beta2- and either gamma1- or gamma2-subunits. Activated AMPK, detected by immunohistochemical staining for phospho-Thr172 AMPK (pThr172), was expressed on the apical surface of the cortical thick ascending limb of the loop of Henle, including the macula densa, and some parts of the distal convoluted tubule. Activated AMPK was also expressed on the basolateral surface of the cortical and medullary collecting ducts as well as some portions of the distal convoluted tubules. AMPK activity was increased by 25% in animals receiving a high-salt diet, and this was confirmed by Western blotting for pThr172. Low-salt diets were associated with reduced levels of the alpha-subunit of AMPK, which was highly phosphorylated on Thr172. Surprisingly, both low- and high-salt media transiently activated AMPK in the macula densa cell line MMDD1, an effect due to changes in osmolality, rather than Na+ or Cl- concentration. This study, therefore, demonstrates regulation of AMPK by both a high- and a low-salt intake in vivo and suggests a role for the kinase in the response to changes in osmolality within the kidney. PMID- 15536170 TI - Integrated actions of transforming growth factor-beta1 and connective tissue growth factor in renal fibrosis. AB - Matrix accumulation in the renal tubulointerstitium is predictive of a progressive decline in renal function. Transforming growth factor-beta(1) (TGF beta(1)) and, more recently, connective tissue growth factor (CTGF) are recognized to play key roles in mediating the fibrogenic response, independently of the primary renal insult. Further definition of the independent and interrelated effects of CTGF and TGF-beta(1) is critical for the development of effective antifibrotic strategies. CTGF (20 ng/ml) induced fibronectin and collagen IV secretion in primary cultures of human proximal tubule cells (PTC) and cortical fibroblasts (CF) compared with control values (P < 0.005 in all cases). This effect was inhibited by neutralizing antibodies to either TGF-beta or to the TGF-beta type II receptor (TbetaRII). TGF-beta(1) induced a greater increase in fibronectin and collagen IV secretion in both PTC (P < 0.01) and CF (P < 0.01) compared with that observed with CTGF alone. The combination of TGF beta(1) and CTGF was additive in their effects on both PTC and CF fibronectin and collagen IV secretion. TGF-beta(1) (2 ng/ml) stimulated CTGF mRNA expression within 30 min, which was sustained for up to 24 h, with a consequent increase in CTGF protein (P < 0.05), whereas CTGF had no effect on TGF-beta(1) mRNA or protein expression. TGF-beta(1) (2 ng/ml) induced phosphorylated (p)Smad-2 within 15 min, which was sustained for up to 24 h. CTGF had a delayed effect on increasing pSmad-2 expression, which was evident at 24 h. In conclusion, this study has demonstrated the key dependence of the fibrogenic actions of CTGF on TGF-beta. It has further uniquely demonstrated that CTGF requires TGF-beta, signaling through the TbetaRII in both PTCs and CFs, to exert its fibrogenic response in this in vitro model. PMID- 15536171 TI - Relative contributions of Ca2+ mobilization and influx in renal arteriolar contractile responses to arginine vasopressin. AB - Experiments addressed the hypothesis that afferent and efferent arterioles differentially rely on Ca2+ influx and/or release from intracellular stores in generating contractile responses to AVP. The effect of Ca2+ store depletion or voltage-gated Ca2+ channel (VGCC) blockade on contractile responsiveness to AVP (0.01-1.0 nM) was assessed in blood-perfused juxtamedullary nephrons from rat kidney. Depletion of intracellular Ca2+ stores by 100 microM cyclopiazonic acid (CPA) or 1 microM thapsigargin treatment increased afferent arteriolar baseline diameter by 14 and 21%, respectively, but did not significantly alter efferent arteriolar diameter. CPA attenuated the contractile response to 1.0 nM AVP by 34 and 55% in afferent and efferent arterioles, respectively (P = 0.013). The impact of thapsigargin on AVP-induced afferent arteriolar contraction (52% inhibition) was also less than its effect on the efferent arteriolar response (88% inhibition; P = 0.046). In experiments probing the role of the Ca2+ influx through VGCCs, 10 microM diltiazem evoked a 34% increase in baseline afferent arteriolar diameter and attenuated the contractile response to 1.0 nM AVP by 45%, without significantly altering efferent arteriolar baseline diameter or responsiveness to AVP. Combined treatment with both diltiazem and thapsigargin prevented AVP-induced contraction of both vascular segments. We conclude that Ca2+ release from the intracellular stores contributes to the contractile response to AVP in both afferent and efferent arterioles but is more prominent in the efferent arteriole. Moreover, the VGCC contribution to AVP-induced renal arteriolar contraction resides primarily in the afferent arteriole. PMID- 15536172 TI - N-ethylmaleimide causes aquaporin-2 trafficking in the renal inner medullary collecting duct by direct activation of protein kinase A. AB - The antidiuretic hormone arginine vasopressin increases the osmotic water permeability of the renal collecting ducts by inducing the shuttling of aquaporin 2 (AQP2) water channels from intracellular vesicles to the apical plasma membrane of the principal cells. This process has been demonstrated to be dependent on the cytoskeleton and protein kinase A (PKA). Previous studies in the toad urinary bladder, a functional homologue of the renal collecting duct, have demonstrated that the sulfhydryl reagent N-ethylmaleimide (NEM) is also able to activate the vasopressin-sensitive water permeability pathway in this tissue. The aim of the present study was to investigate the effects of NEM on AQP2 trafficking in a mammalian system. We show that NEM causes translocation of AQP2 from the cytosol to the plasma membrane in rat inner medullary collecting ducts; like the response to arginine vasopressin, this action was also dependent on an intact cytoskeleton and PKA. This effect is not mediated by cAMP but results from direct activation of PKA by NEM. PMID- 15536173 TI - In vivo imaging of oxidative stress in ischemia-reperfusion renal injury using electron paramagnetic resonance. AB - Oxidative stress during ischemia-reperfusion acute renal failure (IR-ARF) was noninvasively evaluated with in vivo electron paramagnetic resonance (EPR) imaging. Female ICR mice underwent left nephrectomy and 30-min ischemia reperfusion of the right kidney. Oxidative stress was evaluated as organ reducing activity with the half-lives of the spin probe 3-carbamoyl-2,2,5,5 tetramethylpyrrolidine-1-oxyl (carbamoyl-PROXYL) using 1) conventional L-band EPR, which showed organ-reducing activity in the whole abdominal area; and 2) EPR imaging, which showed semiquantitative but organ-specific reducing activity. The results were compared with the reducing activity of organ homogenate and phosphatidylcholine hydroperoxide (PC-OOH) concentrations. Half-lives of carbamoyl-PROXYL in the whole upper abdominal area, measured by L-band EPR, were prolonged on day 3 after ischemia-reperfusion and recovered to the level of nontreated mice on day 7. This trend resembled closely that of serum creatinine and blood urea nitrogen concentration. The EPR imaging-measured carbamoyl-PROXYL half-life was also prolonged on day 3 in both the kidney and the liver. However, in the kidney this showed only partial recovery on day 7. In the liver, this convalescence was more remarkable. The ex vivo studies of organ reducing activity and PC-OOH agreed with the results from EPRI, but not with those from L-band EPR. These results indicate that renal reducing activity shows only partial recovery on day 7 after ischemia-reperfusion, when serum creatinine and blood urea nitrogen have recovered. EPR imaging is an appropriate and useful method for the noninvasive evaluation of oxidative stress in the presence of renal injury. PMID- 15536174 TI - Serial analysis of gene expression during elongation of the peri-implantation porcine trophectoderm (conceptus). AB - Conceptus loss during the preimplantation and early postimplantation period hinders the efficiency of swine reproduction. Significant conceptus loss occurs during trophectoderm elongation between gestational day 11 (D11) and day 12 (D12). Elongation of the porcine conceptus is a key stage of development during which maternal recognition of pregnancy, initial placental development, and preparation for implantation occurs. The objective of this study was to establish comparative transcriptome profiles of D11 ovoid and D12 filamentous conceptuses and thereby identify temporally regulated genes essential for developmental progression during conceptus elongation. Serial analysis of gene expression (SAGE) libraries were constructed from in vivo derived ovoid and filamentous swine conceptuses to yield a total of 42,389 tags (ovoid) and 42,391 tags (filamentous) representing 14,464 and 13,098 putative unique transcripts, respectively. Statistical analysis of tag frequencies revealed the differential expression of 431 tags between libraries (P < 0.05). Nucleotide sequence alignment searches on public databases provided SAGE tag annotation and gene ontology assignments. Comparisons between the SAGE profiles of ovoid and filamentous conceptuses revealed increased expression of key genes in the steroidogenesis [cytochrome P-450(scc) (CYP11A1), aromatase (CYP19A), and steroidogenic acute regulatory protein (STAR)] and oxidative stress response pathways [microsomal glutathione S-transferase 1 (MGST1) and copper-zinc superoxide dismutase (SOD1)]. Differential expression of these genes in the steroidogenic and oxidative stress response pathways was confirmed by real-time PCR. These results validate the utility of SAGE in the pig and establish an initial model linking gene expression profiles at the pathway level with phenotypic progression from ovoid to filamentous stages of conceptus development. PMID- 15536175 TI - Detecting ATP release by a biosensor method. AB - Cells release adenosine 5'-triphosphate (ATP) into the extracellular space in response to various stimuli. This released ATP plays an important physiological role in cell-to-cell signal transduction. The bulk ATP concentration can be detected using a conventional luciferin-luciferase assay. However, the ATP concentration in the vicinity of the cell surface is often different from the bulk concentration because of its rapid degradation by ecto-ATPases and because of delayed diffusion due to unstirred layer effects. Here, we describe a simple biosensor method to measure the local ATP concentration on the cell surface in real time. The method is based on the ATP-dependent opening of ligand-gated cation channels of purinergic P2X receptors expressed in undifferentiated pheochromocytoma (PC12) cells or in human embryonic kidney 293 (HEK293) cells stably transfected with recombinant P2X2 purinergic receptors. Under the whole cell configuration of patch-clamp, a sensor PC12 cell or HEK293 is positioned within the proximity of a target cell, and the P2X-mediated currents induced by ATP released from a given site on the target cell surface is measured. The ATP release is quantified by a calibration procedure utilizing local puff applications of ATP at preset concentrations. PMID- 15536176 TI - Spectral analysis of calcium oscillations. AB - Calcium (Ca2+) oscillations are universal signals exploited by cells to regulate a vast number of cellular processes. Frequency and amplitude, the key features of oscillating waves, can yield information that can be decoded by intracellular processes. Analysis and quantification of Ca2+ oscillations are crucial for understanding the general concept of Ca2+ signaling. This protocol presents a method for performing spectral analysis of Ca2+ oscillations using MATLAB software. PMID- 15536179 TI - Transient expression of Olig1 initiates the differentiation of neural stem cells into oligodendrocyte progenitor cells. AB - In order to develop an efficient strategy to induce the in vitro differentiation of neural stem cells (NSCs) into oligodendrocyte progenitor cells (OPCs), NSCs were isolated from E14 mice and grown in medium containing epidermal growth factor and fibroblast growth factor (FGF). Besides supplementing the medium with oligodendrogenic factors such as Sonic Hedgehog (Shh), FGF-2, and PDGF, we attempted to initiate the gene transcription program for OPC differentiation by transfection of the Olig1 gene, a transcription factor known to be involved in the induction of oligodendrocyte lineage formation during embryogenesis. Whereas addition of Shh, FGF-2, and PDGF could induce OPC differentiation in 12% of the NSCs, the transient expression of Olig1 by use of Nucleofector gene transfection initiated OPC differentiation in 55% of the NSCs. Our results show that nonviral transfection of genes encoding for oligodendrogenic transcription factors may be an efficient way to initiate the in vitro differentiation of NSCs into OPCs. PMID- 15536180 TI - Presence of functional gap junctions in human embryonic stem cells. AB - Gap junctions are intercellular channels that allow both chemical and electrical signaling between two adjacent cells. Gap junction intercellular communication has been implicated in the regulation of various cellular processes, including cell migration, cell proliferation, cell differentiation, and cell apoptosis. This study aimed to determine the presence and functionality of gap junctions in human embryonic stem cells (hESCs). Using reverse transcription--polymerase chain reaction and immunocytochemistry, we demonstrate that human ES cells express two gap junction proteins, connexin 43 and connexin 45. Western blot analysis revealed the presence of three phosphorylated forms (nonphosphorylated [NP], P1, and P2) of connexin 43, NP being prominent. Moreover, scrape loading/dye transfer assay indicates that human ES cells are coupled through functional gap junctions that are inhibited by protein kinase C activation and extracellular signal regulated kinase inhibition. PMID- 15536181 TI - Differential effects of culture conditions on the migration pattern of stromal cell-derived factor-stimulated hematopoietic stem cells. AB - Evidence is mounting that hematopoietic stem cells (HSCs) play a critical role in bone marrow regeneration and tissue renewal, for which migration is an obvious prerequisite. Computer-aided analysis and a three-dimensional collagen matrix assay enabled us to analyze single-cell migratory characteristics of stromal cell derived factor-1 alpha (SDF-1 alpha)-stimulated cord blood-derived HSCs. We defined and resolved specific migratory parameters in spontaneous and SDF-1 alpha induced migration of these cells. The addition of interleukin 6 to the culture medium led to differential SDF-1 alpha-stimulated migratory response, which comprised a recruitment of nonmoving cells and an increase in speed and frequency of pauses but a decrease in pause duration. We were thus able to decipher the exact parameters that result in an increase in the migration of HSCs and demonstrate that extensive analysis of single-cell behavior is elementary in the study of stem cell migration. PMID- 15536182 TI - Potential of hematopoietic stem cell therapy in hepatology: a critical review. AB - Adult stem cell plasticity raised expectations regarding novel cellular therapies of regenerative medicine after findings of unexpected plasticity were reported. In this review, reports of hematopoietic stem cells (HSCs) contributing to hepatocytic lineages are critically discussed with reference to rodent and human models. In particular, the role of liver injury and the potential contribution HSCs make to hepatic regeneration in both injury and physiological maintenance is reviewed. The relative contributions of genomic plasticity and cell fusion are studied across different model systems, highlighting possible factors that may explain differences between often conflicting reports. Insights from experimental studies will be described that shed light on the mechanisms underlying the migration, engraftment, and transdifferentiation of HSCs in liver injury. Although it appears that under differing circumstances, macrophage fusion, HSC fusion, and HSC transdifferentiation can all contribute to hepatic epithelial lineages, a much greater understanding of the factors that regulate the long-term efficacy of such cells is needed before this phenomenon can be used clinically. PMID- 15536183 TI - Fas ligand as a tool for immunosuppression and generation of immune tolerance. AB - The role of Fas ligand (FasL) in physiologically limiting immune responses and maintaining immune-privileged sites has led to a body of research aiming to confer protection to allogeneic grafts by expressing FasL on the allogeneic tissue or by administrating FasL-transduced donor dendritic cells. In addition, several studies have used FasL to abrogate autoimmune responses. This review presents the results of these studies and discusses the problems associated with FasL usage. PMID- 15536184 TI - Dopaminergic differentiation of human embryonic stem cells. AB - In this manuscript we report that human embryonic stem cells (hESCs) differentiated into dopaminergic neurons when cocultured with PA6 cells. After 3 weeks of differentiation, approximately 87% of hES colonies contained tyrosine hydroxylase (TH)-positive cells, and a high percentage of the cells in most of the colonies expressed TH. Differentiation was inhibited by exposure to BMP4 or serum. TH-positive cells derived from hESCs were postmitotic, as determined by bromodeoxyurindine colabeling. Differentiated cells expressed other markers of dopaminergic neurons, including the dopamine transporter, aromatic amino acid decarboxylase, and the transcription factors associated with neuronal and dopaminergic differentiation, Sox1, Nurr1, Ptx3, and Lmx1b. Neurons that had been differentiated on PA6 cells were negative for dopamine-beta-hydroxylase, a marker of noradrenergic neurons. PA6-induced neurons were able to release dopamine and 3,4-dihydroxphe-hylacetic acid (DOPAC) but not noradrenalin when depolarized by high K(+). When transplanted into 6-hydroxydopamine-treated animals, hES-derived dopaminergic cells integrated into the rat striatum. Five weeks after transplantation, surviving TH-positive cells were present but in very small numbers compared with the high frequency of TH-positive cells seen in PA6 coculture. Larger numbers of cells positive for smooth muscle actin, but no undifferentiated ES cells, were present after transplantation. Therefore, hESCs can be used to generate human dopaminergic cells that exhibit biochemical and functional properties consistent with the expected properties of mature dopaminergic neurons. PMID- 15536185 TI - Nuclei of embryonic stem cells reprogram somatic cells. AB - The restricted potential of a differentiated cell can be reverted back to a pluripotent state by cell fusion; totipotency can even be regained after somatic cell nuclear transfer. To identify factors involved in resetting the genetic program of a differentiated cell, we fused embryonic stem cells (ESCs) with neurosphere cells (NSCs). The fusion activated Oct4, a gene essential for pluripotency, in NSCs. To further identify whether cytoplasmic or nuclear factors are responsible for its reactivation, we fused either karyoplasts or cytoplasts of ESCs with NSCs. Our results show that ESC karyoplasts could induce Oct4 expression in the somatic genome, but cytoplasts lacked this ability. In addition, mitomycin C-treated ESCs, although incapable of DNA replication and cell division, could reprogram 5-azacytidine-treated NSCs. We therefore conclude that the Oct4 reprogramming capacity resides in the ESC karyoplast and that gene reactivation is independent of DNA replication and cell division. PMID- 15536186 TI - Effects of type IV collagen and laminin on the cryopreservation of human embryonic stem cells. AB - Previous reports have indicated that extracellular matrices (ECMs) affect the developmental fate of human embryonic stem cells (hESCs). Specially, type IV collagen and laminin, which belong to a group of macromolecular proteins with a substantial proportion of ECMs, are known to influence the proliferation and differentiation of hES cells. In this study, we evaluated the effects of type IV collagen and laminin in freezing medium on the survival and differentiation rates of hES cells after slow freezing and rapid thawing. The addition of type IV collagen (1 microg/ml) to the freezing medium significantly increased the survival rate of hES cells after thawing compared with that of a control group. The spontaneous differentiation rates of groups treated with type IV collagen (1 microg/ml) or laminin (1 microg/ml) were significantly lower than those of the control group. Frozen-thawed hES cells have currently been cultured for more than 70 passages and retain key properties of hES cells such as morphological characteristics, normal karyotype, marker expression (alkaline phosphatase, SSEA 1, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, Rex-1, and Oct-4), basement membrane related gene expression, and the potential to differentiate into derivatives of all three germ layers. This new slow freezing method by ECM treatment is a reliable and effective cryopreservation method for pluripotent hES cells. PMID- 15536187 TI - Stress defense in murine embryonic stem cells is superior to that of various differentiated murine cells. AB - A very small number of embryonic stem (ES) cells gives rise to all tissues of the embryo proper. This means that ES cells should be equipped with highly efficient mechanisms to defend themselves against various stresses and to prevent or repair DNA damage. One of these mechanisms is a high activity of a verapamil-sensitive multidrug efflux pump. Because reactive oxygen species are a major source of DNA damage, we further tested the idea that murine ES cells might differ from their more differentiated counterparts by high levels of antioxidant defense and good DNA strand break repair capacity. This was confirmed by comparing cellular peroxide levels, total antioxidant capacity, and activity of radiation-induced strand break repair between murine ES cells and embryoid bodies or embryonic fibroblasts. Using microarrays and confirmation by reverse transcription polymerase chain reaction, we identified several candidate antioxidant and stress resistance genes that become downregulated during differentiation of ES cells into embryoid bodies. PMID- 15536188 TI - Immortalized fibroblast-like cells derived from human embryonic stem cells support undifferentiated cell growth. AB - Human embryonic stem cells (hESCs) have the potential to generate multiple cell types and hold promise for future therapeutic applications. Although undifferentiated hESCs can proliferate indefinitely, hESC derivatives significantly downregulate telomerase and have limited replication potential. In this study we examine whether the replicative lifespan of hESC derivatives can be extended by ectopic expression of human telomerase reverse transcriptase (hTERT), the catalytic component of the telomerase complex. To this end, we have derived HEF1 cells, a fibroblast-like cell type, differentiated from hESCs. Infection of HEF1 cells with a retrovirus expressing hTERT extends their replicative capacity, resulting in immortal human HEF1-hTERT cells. HEF1-hTERT cells can be used to produce conditioned medium (CM) capable of supporting hESC growth under feeder free conditions. Cultures maintained in HEF1-CM show characteristics similar to mouse embryonic fibroblast CM control cultures, including morphology, surface marker and transcription factor expression, telomerase activity, differentiation, and karyotypic stability. In addition, HEF1-hTERT cells have the capacity to differentiate into cells of the osteogenic lineage. These results suggest that immortalized cell lines can be generated from hESCs and that cells derived from hESCs can be used to support their own growth, creating a genotypically homogeneous system for the culture of hESCs. PMID- 15536189 TI - Human umbilical cord blood cells differentiate into muscle in sjl muscular dystrophy mice. AB - Limb girdle muscular dystrophy type 2B form (LGMD-2B) and Miyoshi myopathy (MM) are both caused by mutations in the dysferlin (dysf) gene. In this study, we used dysferlin-deficient sjl mice as a mouse model to study cell therapy for LGMD-2B and MM. A single-blind study evaluated the therapeutic potential of human umbilical cord blood (HUCB) as a source of myogenic progenitor stem cells. Three groups of donor cells were used: unfractionated mononuclear HUCB cells, HUCB subfractionated to enrich for cells that were negative for lineage surface markers (LIN(-)) and substantially enriched for the CD34 surface marker (CD34(+)), and irradiated control spleen cells. We administrated 1 x 10(6) donor cells to each animal intravenously and euthanized them at different time points (1-12 weeks) after transplantation. All animals were immunosuppressed (FK506 and leflunomide) from the day before the injection until the time of euthanasia. Immunohistochemical analyses documented that a small number of human cells from the whole HUCB and LIN(-)CD34(+/-)-enriched HUCB subgroups engraft in the recipient muscle to express both dysferlin and human-specific dystrophin at 12 weeks after transplantation. We conclude that myogenic progenitor cells are present in the HUCB, that they can disseminate into muscle after intravenous administration, and that they are capable of myogenic differentiation in host muscle. PMID- 15536190 TI - Derivation of functional endothelial progenitor cells from human umbilical cord blood mononuclear cells isolated by a novel cell filtration device. AB - Endothelial progenitor cells (EPCs) can differentiate from mononuclear cells (MNCs) of adult human peripheral blood, bone marrow, and cord blood during culture. Although MNCs are usually isolated by a Ficoll gradient centrifuge method, this method is time-consuming, and blood is easily contaminated. We developed a novel cell filtration device (StemQuickE, Asahi Kasei Medical, Oita, Tokyo, Japan) to isolate MNCs from human cord blood and examined whether functional EPCs could differentiate from MNCs isolated by this device. Recovery rates of MNCs, CD34(+) and CD133(+) progenitor cells, were significantly greater in the StemQuickE method than in the Ficoll method. During MNC culture, spindle shaped attaching cells developed, and most of these cells incorporated DiI acetylated low-density lipoprotein and showed positive binding to fluorescein isothiocyanate-lectin. Reverse transcription-polymerase chain reaction analysis revealed that attaching cells expressed various progenitor and endothelial lineage markers such as KDR, CD31, endothelial cell nitric oxide synthase, CD133, and LOX-1. Culture-expanded EPCs were isolated and labeled with a green fluorescent dye, PKH2-GL, and cocultured with human umbilical vein endothelial cells (HUVECs). EPCs formed angiogenesis-like networks together with HUVECs in 3D matrix gel. Finally, EPCs (3 x 10(5)) were implanted into ischemic hindlimb of nude rats (n = 3), and laser Doppler blood flowmetry (LDBF) revealed that the ratio of ischemic to normal limb LDBF was significantly greater in EPC transplanted animals compared with controls receiving saline. In conclusion, the novel cell filtration device, StemQuickE, is a useful tool to isolate MNCs from human cord blood. Moreover, MNCs obtained by this filter system can give rise to functional EPCs. PMID- 15536191 TI - Comparative proteomic analysis of human CD34+ stem/progenitor cells and mature CD15+ myeloid cells. AB - Human CD34(+) cells, highly enriched for hematopoietic stem and progenitors, and CD15(+) cells, more terminally differentiated myeloid cells in blood, represent distinct maturation/differentiation stages. A proteomic approach was used to identify proteins differentially present in these two populations from human cord blood. Cytosolic proteins were extracted and subjected to two-dimensional gel electrophoresis followed by mass spectrometry. On average, 460 protein spots on each gel were detected; 112 and 15 proteins, respectively, were found to be differentially expressed or post-translationally modified in CD34(+) and CD15(+) cells. This suggests that CD34(+) cells have a relatively larger proteome than mature CD15(+) myeloid cells and production of many stem/progenitor cell associated proteins ceases or is dramatically down-regulated as the CD34(+) cells undergo differentiation. Of approximately 140 protein spots, 47 different proteins were positively identified by mass spectrometry and database search; these proteins belong to several functional categories, including cell signaling, transcription factors, cytoskeletal proteins, metabolism, protein folding, and vesicle trafficking. Multiple heat shock proteins and chaperones, as well as proteins important for intracellular trafficking, were predominantly present in CD34(+) cells. Most of the identified proteins in CD34(+) cells are expressed in germ cell tumors, as well as in embryonal carcinoma and neuroblastoma. Approximately eight novel proteins, whose functions are unknown, were identified. This study presents, for the first time, global cellular protein expression patterns in human CD34(+) and CD15(+) cells, which should help to better understand intracellular processes involved in myeloid differentiation and add insight into the functional capabilities of these distinct cell types. PMID- 15536192 TI - Intracellular localization and constitutive endocytosis of CXCR4 in human CD34+ hematopoietic progenitor cells. AB - CXCR4, the stromal cell-derived factor-1 receptor, plays an important role in the migration of hematopoietic progenitor/stem cells. The surface and cytoplasmic expression of CXCR4 on human hematopoietic CD34(+) cells was investigated. We show that its surface expression is low, whereas a large part of CXCR4 protein is sequestered intracellularly. Using confocal microscopy, we demonstrated that CXCR4 is colocalized with EEA-1, Rab5, Rab4, and Rab11, which are localized in early and recycling endosomes. No significant colocalization of CXCR4 with lysosomal markers CD63 and Lamp-1 was detected. Using antibody feeding experiments, we report a role for CXCR4 constitutive endocytosis in subcellular localization in stably transduced UT7-CXCR4-GFP and CD34(+) cells. Agonist independent endocytosis of CXCR4 occurs through clathrin-coated vesicles. These data implicate a constitutive endocytosis in the regulation of CXCR4 membrane expression and suggest that constitutive endocytosis may be involved in the regulation of trafficking the human hematopoietic progenitor/stem cells to and in the bone marrow microenvironment. PMID- 15536193 TI - A specific heptapeptide from a phage display peptide library homes to bone marrow and binds to primitive hematopoietic stem cells. AB - Phage display peptide libraries have enabled the discovery of peptides that selectively target specific organs. Selection of organ-specific peptides is mediated through binding of peptides displayed on phage coat protein to adhesion molecules expressed within targeted organs. Hematopoietic stem cells selectively home to bone marrow, and certain adhesion receptors critical to this function have been demonstrated. Using a phage display library, we identified a specific peptide that trafficked to murine bone marrow in vivo. We independently isolated exactly the same heptapeptide from the entire library by in vitro biopanning on primitive lineage-depleted, Hoechst 33342(dull)/rhodamine 123(dull) murine bone marrow stem cells and confirmed peptide binding to these cells by immunofluorescence studies. We demonstrated bone marrow-specific homing of the peptide by an in vivo assay in which the animals were injected with the phage displaying peptide sequence, and immunofluorescence analysis of multiple organs was performed. We also showed that the peptide significantly decreased the homing of stem cells to the bone marrow but not to the spleen 3 hours after transplantation using fluorescently labeled Lin(-)Sca(+) hematopoietic cells in an in vivo homing assay. The peptide sequence has a partial (5/7) amino acid sequence homology with a region of CD84. This discovery represents the first application of the phage display methodology to the bone marrow and stem cells and led to the identification of a specific heptapeptide that homes to bone marrow, binds to primitive stem cells, and plays a role in stem cell homing. PMID- 15536194 TI - CD45 congenic bone marrow transplantation: evidence for T cell-mediated immunity. AB - CD45 congenic mice have been used to study stem cell engraftment in the absence of alloreactivity. Recently, impaired engraftment was reported in this model and attributed to weak immune reactivity. We have confirmed that there is indeed low level reactivity mediated by CD8(+) cells and alpha beta-TCR(+) T cells. B6 (CD45.2) recipients were conditioned with total body irradiation (TBI) and transplanted with increasing doses of B6 (CD45.1) bone marrow cells (BMCs). Although chimerism was present at 1 month in all recipients, durable engraftment did not occur with <150 cGy of TBI, emphasizing the importance of long-term follow-up in evaluating nonmyeloablative conditioning approaches. A single dose of cyclophosphamide on day 2 also significantly enhanced engraftment. When B6 TCR beta(-/-), TCR delta(-/-), or TCR beta(-/-)/delta(-/-) (CD45.2) mice were transplanted with CD45.1 bone marrow, significantly enhanced engraftment occurred in recipients lacking alpha beta-TCR(+) T cells (p < .00005). Similarly, removal of alpha beta-TCR(+) host T cells in wild-type recipients resulted in enhanced engraftment. Engraftment was also significantly increased in CD8(-/-) and CD4(-/ )/8(-/-) recipients (p < .0005). Taken together, these results demonstrate that alpha beta-TCR(+) and CD8(+) T cells play a critical role in regulating engraftment of CD45 congenic marrow and suggest that these cells are the main effector cells in low-level alloreactivity to the CD45 disparity. PMID- 15536195 TI - Bone marrow progenitors are not the source of expanding oval cells in injured liver. AB - Liver progenitor/oval cells differentiate into hepatocytes and biliary epithelial cells, repopulating the liver when the regenerative capacity of hepatocytes is impaired. Recent studies have shown that hematopoietic bone marrow (BM) stem/progenitor cells can give rise to hepatocytes in diseased/damaged liver. One study has reported that BM cells can transdifferentiate into liver progenitor/oval cells, but it has not been proven that the latter can repopulate the liver. To answer this question, we have lethally irradiated female DPP4(-) mutant F344 rats and transplanted them with 50 million wild-type male F344 BM cells. One month after transplantation, the recipient BM was reconstituted with male hematopoietic cells, determined by quantitative polymerase chain reaction using primers for Y chromosome-specific sry gene. In addition, DPP4(+) cells, single or in clusters and predominantly in the periportal region, were detected in all liver sections of recipient rats. Animals were subjected to the following three different liver injury protocols for activation and expansion of oval cells: D-galactosamine, retrorsine/partial hepatectomy (Rs/PH), and 2 acetylaminofluorene/partial hepatectomy (2-AAF/PH). In all three models, prominent expansion and accumulation of cytokeratin 19-positive (CK-19(+)) oval cells was observed. However, most of the DPP4(+) clusters dispersed over time, and their total number decreased. Very few oval cells (less than 1%) showed double DPP4/CK-19 labeling. None of the small hepatocytic clusters in the Rs/PH or 2-AAF/PH model were comprised of DPP4(+) cells. These data demonstrate that the sources of oval cells and small hepatocytes in the injured liver are endogenous liver progenitors and that they do not arise through transdifferentiation from BM cells. PMID- 15536196 TI - Comparison of retroviral transduction efficiency in CD34+ cells derived from bone marrow versus G-CSF-mobilized or G-CSF plus stem cell factor-mobilized peripheral blood in nonhuman primates. AB - Hematopoietic stem cells (HSCs) are ideal targets for genetic manipulation in the treatment of several congenital and acquired disorders affecting the hematopoietic compartment. Although G-CSF-mobilized peripheral blood CD34(+) cells are the favored source of hematopoietic stem cells in clinical transplantation, this source of stem cells does not provide meaningful engraftment levels of genetically modified cells compared with G-CSF + stem cell factor (SCF)-mobilized cells in nonhuman primates. Furthermore, the use of G-CSF mobilization can have disastrous consequences in patients with sickle cell disease, a long-held target disorder for HSC-based gene therapy approaches. We therefore conducted a study to compare the levels of genetically modified cells attainable after retroviral transduction of CD34(+) cells collected from a bone marrow (BM) harvest with CD34(+) cells collected from a leukapheresis product after mobilization with G-CSF (n = 3) or G-CSF in combination with SCF (n = 3) in the rhesus macaque autologous transplantation model. Transductions were performed using retroviral vector supernatant on fibronectin-coated plates for 96 hours in the presence of stimulatory cytokines. BM was equal to or better than G-CSF mobilized peripheral blood as a source of HSCs for retroviral transduction. Although the highest marking observed was derived from G-SCF + SCF-mobilized peripheral blood in two animals, marking in the third originated only from the BM fraction. These results demonstrate that steady-state BM is at least equivalent to G-CSF-mobilized peripheral blood as a source of HSCs for retroviral gene transfer and the only currently available source for patients with sickle cell disease. PMID- 15536197 TI - Adult pancreas generates multipotent stem cells and pancreatic and nonpancreatic progeny. AB - Strategies designed to produce functional cells from stem cells or from mature cells hold great promise for treatment of different cell-degenerative diseases. Type 1 and type 2 diabetes are examples of such diseases. Although different in origin, both involve inadequate cell mass of insulin-producing beta cells, the most abundant cell type of pancreatic islets of Langerhans. Practical realization of such strategies is highly dependent on the elucidation of physiological mechanisms responsible for generation of new beta cells in the pancreas, which at this time are poorly defined. The in vitro differentiation systems allowing generation of new beta cells provide a valuable experimental tool for studying these mechanisms. Few such systems are currently available. In this work, we present an in vitro differentiation system, derived from adult mouse pancreas, capable of generating insulin-producing beta-like cells, which self-organize into islet-like cell clusters (ILCCs) during the course of the culture. Surprisingly, we found that along with the ILCCs, multiple cell types with phenotypic characteristics of embryonic central nervous system and neural crest are also generated. Moreover, several embryonic stem cell-specific genes are induced during the course of these cultures. These results suggest that the adult pancreas may contain cells competent to give rise to new endocrine and neural cells. PMID- 15536198 TI - Tumor necrosis factor promotes human T-cell development in nonobese diabetic/severe combined immunodeficient mice. AB - A major problem after clinical hematopoietic stem cell transplantations is poor T cell reconstitution. Studying the mechanisms underlying this concern is hampered, because experimental transplantation of human stem and progenitor cells into nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice usually results in low T-lymphocyte reconstitution. Because tumor necrosis factor alpha (TNFalpha) has been proposed to play a role in T-lineage commitment and differentiation in vitro, we investigated its potential to augment human T-cell development in vivo. Administration of TNF to irradiated NOD/SCID mice before transplantation of human mononuclear cells from either cord blood or adult G-CSF mobilized peripheral blood (MPBL) led 2-3 weeks after transplantation to the emergence of human immature CD4(+)CD8(+) double-positive T-cells in the bone marrow (BM), spleen, and thymus, and in this organ, the human cells also express CD1a marker. One to 2 weeks later, single-positive CD4(+) and CD8(+) cells expressing heterogenous T-cell receptor alpha beta were detected in all three organs. These cells were also capable of migrating through the blood circulation. Interestingly, human T-cell development in these mice was associated with a significant reduction in immature lymphoid human CD19(+) B cells and natural killer progenitors in the murine BM. The human T cells were mostly derived from the transplanted immature CD34(+) cells. This study demonstrates the potential of TNF to rapidly augment human T lymphopoiesis in vivo and also provides clinically relevant evidence for this process with adult MPBL progenitors. PMID- 15536199 TI - Absence of major histocompatibility complex class I on neural stem cells does not permit natural killer cell killing and prevents recognition by alloreactive cytotoxic T lymphocytes in vitro. AB - Potential applications of neural stem cells (NSCs) for transplantation requires understanding myosin heavy chain (MHC) expression and the ability of T cells and natural killer (NK) cells to recognize this progenitor population. Cells from the cortices of day-13 embryonic (E13) B6 (H-2(b)) mice were explanted and cultured to expand NSCs. Analysis of P2-P17-cultured cells using anti-MHC class I/II monoclonal antibodies (mAbs) showed marginal expression of both products. Although recombinant murine interferon-gamma (rmIFN gamma) exposure did not alter the multipotential capacity of these stem cells, titration of mrIFN gamma NSC cultures demonstrated that MHC molecules could be strongly upregulated after addition of 3 ng/ml rmIFN gamma for 60 hours. To assess the susceptibility of NSCs with low or absent versus high levels of MHC expression to lysis by cytotoxic T lymphocyte (CTL) and NK populations, untreated and rmIFN gamma treated NSC target cells were examined. Untreated NSCs were not recognized by BALB/c (H-2(d)) allospecific anti-H-2(b) CTL, consistent with the mAb findings; however, upregulation of MHC products on both early and later passaged NSCs resulted in their efficient lysis by CTL. NK cells were prepared from syngeneic B6 or allogeneic BALB/c mice. Although NK cells effectively killed control YAC-1 target cells, these effectors did not kill MHC-deficient (or expressing) NSC targets. Thus, similar to hematopoietic, embryonic, and mesenchymal stem cell populations, unmanipulated NSCs are not readily killed by T and NK cells. These findings suggest that following transplant into syngeneic or allogeneic recipients, NSCs may exhibit diminished susceptibility to clearance by host T- and NK-cell populations. PMID- 15536200 TI - Cell division rates of primary human precursor B cells in culture reflect in vivo rates. AB - Bone marrow stroma-based cultures provide a powerful model for studying cell division and apoptosis of primary human precursor B cells. Studies using this model are elucidating the mechanisms by which stromal cells inhibit apoptosis of cultured normal precursor B cells and have demonstrated that the apoptotic rate of cultured leukemic precursor B cells can predict clinical outcome in acute lymphoblastic leukemia. In contrast to apoptosis, cell division in this model has not been well characterized. In this study, we quantified the rates of cell division in cultured primary human normal and leukemic precursor B cells by labeling precursor B cells with the fluorescent dye carboxyfluorescein diacetate, succinimyl ester. Based on the rate of decreasing fluorescent signal over 3 weeks, normal CD19(+), CD10(+) precursor B cells divided once every 90.5 hours, a number that correlates well with the known in vivo rate of 65.5 hours. The division rates were similar among different cultures and constant throughout the 3 weeks of culture, suggesting that the variable expansions of precursor B cells seen among different samples and culture durations are not secondary to different cell division rates. Unlike normal cells, cultured leukemic B cells had a heterogeneous division rate that ranged from once every 26-240 hours. These rates correlated well with their respective in vivo proliferation index. These findings indicate that the stroma-based cultures faithfully replicate in vivo cell division rates and can be used to elucidate the pathways that regulate cell division of primary human precursor B cells. PMID- 15536201 TI - The molecular perspective: cyclins. PMID- 15536202 TI - Isolating bone marrow stem cells using sieve technology. PMID- 15536203 TI - Comparative effects of DHEA vs. testosterone, dihydrotestosterone, and estradiol on proliferation and gene expression in human LNCaP prostate cancer cells. AB - Serum levels of the adrenal androgen dehydroepiandrosterone (DHEA) peak in men and women in the third decade of life and decrease progressively with age. Increasing numbers of middle-aged and older individuals consume over-the-counter preparations of DHEA, hoping it will retard aging by increasing muscle and bone mass and strength, decreasing fat, and improving immunologic and neurobehavioral functions. Because DHEA can serve as a precursor to more potent androgens and estrogens, like testosterone (T), dihydrotestosterone (DHT), and 17beta-estradiol (E2), supplemental DHEA use may pose a cancer risk in patients with nascent or occult prostate cancer. The steroid-responsive human LNCaP prostate cancer cells, containing a functional but mutated androgen receptor (AR), were used to compare effects of DHEA with those of T, DHT, and E2 on cell proliferation and protein and/or gene expression of AR, prostate-specific antigen (PSA), IGF-I, IGF-I receptor (IGF-IR), IGF-II, IGF-binding proteins-2, -3, and -5, (IGFBPs-2, -3, and -5), and estrogen receptor-beta (ERbeta). Cell proliferation assays revealed significant stimulation by all four steroids. DHEA- and E2-induced responses were similar but delayed and reduced compared with that of T and DHT. All four hormones increased gene and/or protein expression of PSA, IGF-IR, IGF-I, and IGFBP-2 and decreased that of AR, ERbeta, IGF-II, and IGFBP-3. There were no significant effects of hormone treatment on IGFBP-5 mRNA. DHEA and E2 responses were similar, and distinct from those of DHT and T, in time- and dose-dependent studies. Further studies of the mechanisms of DHEA effects on prostate cancer epithelial cells of varying AR status, as well as on prostate stromal cells, will be required to discern the implications of DHEA supplementation on prostatic health. PMID- 15536204 TI - Hepatic glycogen breakdown is implicated in the maintenance of plasma mannose concentration. AB - D-mannose is an essential monosaccharide constituent of glycoproteins and glycolipids. However, it is unknown how plasma mannose is supplied. The aim of this study was to explore the source of plasma mannose. Oral administration of glucose resulted in a significant decrease of plasma mannose concentration after 20 min in fasted normal rats. However, in fasted type 2 diabetes model rats, plasma mannose concentrations that were higher compared with normal rats did not change after the administration of glucose. When insulin was administered intravenously to fed rats, it took longer for plasma mannose concentrations to decrease significantly in diabetic rats than in normal rats (20 and 5 min, respectively). Intravenous administration of epinephrine to fed normal rats increased the plasma mannose concentration, but this effect was negated by fasting or by administration of a glycogen phosphorylase inhibitor. Epinephrine increased mannose output from the perfused liver of fed rats, but this effect was negated in the presence of a glucose-6-phosphatase inhibitor. Epinephrine also increased the hepatic levels of hexose 6-phosphates, including mannose 6 phosphate. When either lactate alone or lactate plus alanine were administered as gluconeogenic substrates to fasted rats, the concentration of plasma mannose did not increase. When lactate was used to perfuse the liver of fasted rats, a decrease, rather than an increase, in mannose output was observed. These findings indicate that hepatic glycogen is a source of plasma mannose. PMID- 15536205 TI - Topiramate is an insulin-sensitizing compound in vivo with direct effects on adipocytes in female ZDF rats. AB - We have studied the in vivo and in vitro effects of Topiramate (TPM) in female Zucker diabetic fatty (ZDF) rats. After weight matching, drug treatment had a marked effect to lower fasting glucose levels of relatively normoglycemic animals as well as during an oral glucose tolerance test. The glucose clamp studies revealed a approximately 30% increased glucose disposal, increased hepatic glucose output (HGO) suppression from approximately 30 to 60%, and an increased free fatty acid suppression from 40 to 75%. Therefore, TPM treatment led to enhanced insulin sensitivity at the level of tissue glucose disposal (increased ISGDR), liver (increased inhibition of HGO), and adipose tissue (enhanced suppression of lipolysis). When soleus muscle strips of control or TPM-treated ZDF rats were studied ex vivo, insulin-stimulated glucose transport was not enhanced in the drug-treated animals. In contrast, when isolated adipocytes were studied ex vivo, a marked increase (+55%) in insulin-stimulated glucose transport was observed. In vitro treatment of muscle strips and rat adipocytes showed no effect on glucose transport in muscle with a 40% increase in insulin-stimulated adipocyte glucose transport. In conclusion, 1) TPM treatment leads to a decrease in plasma glucose and increased in vivo insulin sensitivity; 2) insulin sensitization was observed in adipocytes, but not muscle, when tissues were studied ex vivo or in vitro; and 3) TPM directly enhances insulin action in insulin-resistant adipose cells in vitro. Thus the in vivo effects of TPM treatment appear to be exerted through adipose tissue. PMID- 15536206 TI - Burn injury impairs insulin-stimulated Akt/PKB activation in skeletal muscle. AB - The molecular bases underlying burn- or critical illness-induced insulin resistance still remain unclarified. Muscle protein catabolism is a ubiquitous feature of critical illness. Akt/PKB plays a central role in the metabolic actions of insulin and is a pivotal regulator of hypertrophy and atrophy of skeletal muscle. We therefore examined the effects of burn injury on insulin stimulated Akt/PKB activation in skeletal muscle. Insulin-stimulated phosphorylation of Akt/PKB was significantly attenuated in burned compared with sham-burned rats. Insulin-stimulated Akt/PKB kinase activity, as judged by immune complex kinase assay and phosphorylation status of the endogenous substrate of Akt/PKB, glycogen synthase kinase-3beta (GSK-3beta), was significantly impaired in burned rats. Furthermore, insulin consistently failed to increase the phosphorylation of p70 S6 kinase, another downstream effector of Akt/PKB, in rats with burn injury, whereas phosphorylation of p70 S6 kinase was increased by insulin in controls. The protein expression of Akt/PKB, GSK-3beta, and p70 S6 kinase was unaltered by burn injury. However, insulin-stimulated activation of ERK, a signaling pathway parallel to Akt/PKB, was not affected by burn injury. These results demonstrate that burn injury impairs insulin-stimulated Akt/PKB activation in skeletal muscle and suggest that attenuated Akt/PKB activation may be involved in deranged metabolism and muscle wasting observed after burn injury. PMID- 15536207 TI - Beyond the signal sequence: protein routing in health and disease. AB - Receptors, hormones, enzymes, ion channels, and structural components of the cell are created by the act of protein synthesis. Synthesis alone is insufficient for proper function, of course; for a cell to operate effectively, its components must be correctly compartmentalized. The mechanism by which proteins maintain the fidelity of localization warrants attention in light of the large number of different molecules that must be routed to distinct subcellular loci, the potential for error, and resultant disease. This review summarizes diseases known to have etiologies based on defective protein folding or failure of the cell's quality control apparatus and presents approaches for therapeutic intervention. PMID- 15536208 TI - The impact of cost-sharing on appropriate utilization and health status: a review of the literature on seniors. AB - This article provides a review of research that has addressed the impact of patient cost sharing on the use of services and resulting health status impacts, among the population age 65 and older. Nearly all of the 22 relevant studies examined that have been published since 1990-16 focusing on cost-sharing for prescription drugs and 6 on cost-sharing for medical services--conclude that increased cost-sharing reduces either or both the utilization and health status of seniors. Most of the studies, however, rely on cross-sectional and self reported data. Further research, employing stronger study designs as well as clinical and administrative data, is necessary before drawing more definitive conclusions. PMID- 15536209 TI - Translating research into practice: are physicians following evidence-based guidelines in the treatment of hypertension? AB - Despite the widespread availability of evidence-based guidelines for treating hypertension, recent evidence suggests that physicians may not be prescribing first-line drugs for their patients with high blood pressure. Using administrative claims data from 1998 through 2000, this study investigates whether drug treatment provided to 6,736 hypertensives in a privately insured, non-HMO population follows practice guidelines. The authors also examine physician and patient-related factors associated with guideline adherence in a subset of patients with newly diagnosed hypertension. Among members with high blood pressure alone, only 38 percent were on a diuretic, while less than a third were prescribed a beta-blocker, the JNC VI recommended first-line antihypertensives for essential hypertension. Approximately half of individuals with high blood pressure and certain comorbidities received non-first-line interventions. Such findings indicate the need to reconsider how guidelines are communicated and shared with medical practitioners and patients, particularly in light of the drug industry's promotion of newer, more expensive drugs. PMID- 15536210 TI - Low-Income workers with employer-sponsored insurance: who's at risk when employer coverage is no longer an option? AB - A firm's decision to drop the offer of employer-sponsored insurance (ESI), reduce eligibility for ESI, or significantly increase employee costs would have serious implications for the health insurance status of currently covered low-income workers. The authors find that at least a third of currently covered low-income workers do not have affordable insurance options outside of the group market. Furthermore, a simulation analysis shows that 54 percent of those workers would become uninsured if their employers were to drop ESI. This would result in an additional 1 million uninsured adults if 10 percent of low-income workers lost their ESI offer, and at least 350,000 uninsured adults if 10 percent of workers in firms with fewer than 100 employees (the firms most likely to drop coverage) lost their ESI. The authors also find that expanding public programs to cover low income workers would reduce the high uninsurance rate by half, but substantial minorities would remain uninsured. PMID- 15536211 TI - Value for taxpayers' dollars: what VA care would cost at medicare prices. AB - Critics charge that Veterans Health Administration (VA) medical centers are inefficient and the cost of veteran health care would be reduced if VA purchased care for its patients directly from private-sector providers. This analysis compares VA medical care expenditures with estimates of total payments under a hypothetical Medicare fee-for-service payment system reimbursing providers for the same counts of each service VA medical centers provided in fiscal 1999. At six study sites, hypothetical payments were more than 20 percent greater than actual budgets. Nationally, this represented more than 3 billion US dollars in 1999 and more than 5 billion US dollars in 2003. Data limitations suggest the estimate is conservative. Less than half of the difference is due to VA's low pharmacy costs. The study demonstrates the potential savings to patients and taxpayers of the VA health care system. PMID- 15536216 TI - Psychological considerations in pediatric weight management. PMID- 15536217 TI - Preventing and treating obesity in girls and young women to curb the epidemic. AB - Obesity and its serious comorbidities, type 2 diabetes, coronary heart disease, hypertension, and dyslipidemia, have reached epidemic proportions in adults and children. Female obesity is more prevalent and, thus, has greater epidemiological importance: mothers transmit the disease epigenetically and genetically. Maternal obesity affects maternal health, pregnancy outcome, and fetal, neonatal, childhood, and ultimately adult morbidity and mortality. Obesity is easy to diagnose, as are most of its risk factors, yet very little progress has been made in preventing the disease. During a brief period of rapid early growth, there is imprinting of antecedents of adult obesity and obesity-related disease. Because of the rapidity of this early growth and the relative brevity of the critical period, early recognition and prompt intervention are necessary and possibly sufficient to prevent the development of obesity. Identification of inappropriate rapid weight gain through frequent weighing should trigger immediate adjustment of energy intake, a simple intervention in bottle-fed infants, the ones at greatest risk for becoming obese. This review presents a step-care strategy with fail-safe action levels starting with maternal education and diet, exercise, and behavior modification for mother and child and progressing to drug treatment and, in selected cases, laparoscopic surgery for young women of childbearing age in whom other measures have failed. This approach is predicated on the assumption that careful monitoring and responsive supplementation of potential deficiencies is easier to achieve, more cost-effective, and safer than effectively treating manifest obesity and its comorbidities in adults. PMID- 15536218 TI - Obesity research in Canada: literature overview of the last 3 decades. AB - OBJECTIVE: To identify research published on obesity in Canada, to explore the range of areas studied, and to identify gaps and areas that merit future research attention. RESEARCH METHODS AND PROCEDURES: Medline and International Pharmaceutical Abstracts databases were searched from 1970 onwards. Original articles were identified and categorized by areas of interest. RESULTS: A total of 1186 relevant articles were identified: 17, 136, 687, and 346 articles during the 1970s, 1980s, 1990s, and 2000 to 2003, respectively. Of the articles, 816 were considered original studies and accepted for this analysis. Twelve research areas were identified: basic science involving animal experiments (29%), human experiments (16%), populations surveys (14%), obesity-related comorbidities (13%), diagnostic/surgical issues (11%), nonpharmacological approaches (7%), drug related issues (4%), anthropometrics (2%), impact of weight loss (2%), cost/healthcare use (1%), attitudes/perceptions (0.9%), and models/procedures (0.5%). Two-thirds of all research was conducted in Quebec (34%) and Ontario (33%). DISCUSSION: Given the multifactorial nature of obesity, Canadian obesity research covers a broad range of areas with a predominance of basic science but lesser emphasis on community and primary care studies. Furthermore, there was a paucity of research on either clinical management of medical conditions in obese patients or clinical aspects that go beyond weight loss. Thus, although Canada appears well represented in basic research, more attention to exploration of clinical issues and healthcare delivery for obese patients appears warranted. PMID- 15536219 TI - Do psychosocial variables predict weight loss or mental health after obesity surgery? A systematic review. AB - OBJECTIVE: The objective of this study was to present a systematic review of psychological and psychosocial predictors of weight loss and mental health after bariatric surgery. This systematic review included all controlled and noncontrolled trials of the last 2 decades with either a retrospective or prospective design and a follow-up period of at least 1 year. RESEARCH METHODS AND PROCEDURES: The relevant literature was identified by a search of computerized databases. All articles published in English and German between 1980 and 2002 were reviewed. RESULTS: Using the above inclusion/exclusion criteria, 29 articles were identified focusing on psychosocial predictors of weight loss and mental health after obesity surgery. DISCUSSION: Personality traits have no predictive value for the postoperative course of weight or mental state. Apart from serious psychiatric disorders including personality disorders, psychiatric comorbidity seems to be of more predictive value for mental and physical well being as two essential aspects of quality of life than for weight loss postsurgery. However, depressive and anxiety symptoms as correlates of psychological stress with regard to obesity seem to be positive predictors of weight loss postsurgery. The severity of the symptoms or the disorder is more relevant for the outcome of obesity surgery than the specificity of the symptoms. It is also not solely the consumption of distinct "forbidden" foods, such as sweets or soft drinks, but rather a general hypercaloric eating behavior, either as an expression of the patient's inadequate compliance or a dysregulation in energy balance, which is associated with a poor weight loss postsurgery. PMID- 15536220 TI - Molecular screening of the 11beta-HSD1 gene in men characterized by the metabolic syndrome. AB - Adipose tissue type 1 11beta-hydroxysteroid dehydrogenase (11beta-HSD1), which generates hormonally active cortisol from inactive cortisone, has been shown to play a central role in adipocyte differentiation and abdominal obesity-related metabolic complications. The objective was to investigate whether genetic variations in the human 11beta-HSD1 gene are associated with the metabolic syndrome among French-Canadian men. We sequenced all exons, the exon-intron splicing boundaries, and 5' and 3' regions of the human 11beta-HSD1 gene in 36 men with the metabolic syndrome, as defined by the National Cholesterol Education Program-Adult Treatment Panel III, and two controls. Three intronic sequence variants were identified: two single-nucleotide polymorphisms in intron 3 (g.4478T>G) and intron 4 (g.10733G>C) and one insertion in intron 3 (g.4437 4438insA). The relative allele frequency was 19.6%, 22.1%, and 19.6% for the g.4478G, g.10733C, and g.4438insA alleles, respectively. One single-nucleotide polymorphism was identified in exon 6 (c.744G>C or G248G). The frequency of the c.744C allele was only 0.46% in a sample of 217 men. Variants were not associated with components of the metabolic syndrome except for plasma apolipoprotein B levels. In conclusion, molecular screening of the 11beta-HSD1 gene did not reveal any sequence variations that can significantly contribute to the etiology of the metabolic syndrome among French-Canadians. PMID- 15536221 TI - Psychological adjustment of obese youth presenting for weight management treatment. AB - OBJECTIVE: To determine the prevalence of psychological maladjustment in clinic based treatment-seeking obese children and adolescents (BMI > or = 95th percentile) and the degree to which maternal, demographic, and youth factors correlate to the youths' psychological adjustment. RESEARCH METHODS AND PROCEDURES: Anthropometrics, demographics (race, sex, insurance status), measures of youth psychological adjustment (self- and mother-report; Behavior Assessment System for Children), and maternal self-report of psychological distress (Symptom Checklist 90-Revised) were collected from 121 obese children and adolescents (55% white, 45% black) and their mothers. RESULTS: Approximately one-third of youths self-reported some psychological maladjustment, but two-thirds of youth were described by their mothers as experiencing some degree of psychological maladjustment. Adjustment difficulties were specific to social functioning, low self-esteem, and internalizing symptoms. Forty-one percent of mothers of child participants and 56% of mothers of adolescent participants reported clinically significant psychological distress. Youth self-report and mother-report of youths' psychological difficulties were often most strongly associated with mothers' level of psychological distress and/or family socioeconomic status rather than to youth characteristics (e.g., percent overweight, race). DISCUSSION: Psychological maladjustment levels among obese youth and their mothers were higher in this clinic-based sample than in treatment research-based samples. Present correlate findings extended to obese adolescents and reaffirm a potent association between youth self-report of their own psychological adjustment and their mother's level of psychological distress. Demographic characteristics and youth weight status were not consistent correlates of youths' psychological functioning. Findings have implications for the translation of empirically supported pediatric obesity interventions to clinic-based treatment samples. PMID- 15536222 TI - Bariatric surgery patients' views of their physicians' weight-related attitudes and practices. AB - OBJECTIVE: A prior study found that nearly 80% of bariatric surgery patients felt that they were treated disrespectfully by members of the medical profession. This study assessed patient-physician interactions in a group of bariatric surgery patients and in a group of less obese patients who sought weight loss by other means. RESEARCH METHODS AND PROCEDURES: A total of 105 bariatric surgery candidates (mean BMI, 54.8 kg/m(2)) and 214 applicants to a randomized controlled trial of the effects of behavior modification and sibutramine (mean BMI, 37.8 kg/m(2)) completed a questionnaire that assessed patient-physician interactions concerning weight. RESULTS: Only 13% of bariatric surgery patients reported that they were usually or always treated disrespectfully by members of the medical profession, a percentage substantially lower than that found in the previous study. Surprisingly, surgery patients were significantly more satisfied than nonsurgery patients with the care they received for their obesity. Surgery patients also reported significantly more interactions with physicians concerning obesity and weight loss compared with nonsurgery patients. A substantial percentage of both groups, however, reported that their physician did not discuss weight control with them. DISCUSSION: These and other findings suggest that doctor-patient interactions concerning weight may have improved in the past decade; however, there is still much room for improvement. Increased efforts are needed to help physicians discuss, assess, and potentially treat obesity in primary care practice. PMID- 15536223 TI - S 23521 decreases food intake and body weight gain in diet-induced obese rats. AB - OBJECTIVE: To investigate the effect of S 23521, a new glucagon-like peptide-1-(7 36) amide analogue, on food intake and body weight gain in obese rats, as well as on gene expression of several proteins involved in energy homeostasis. RESEARCH METHODS AND PROCEDURES: Lean and diet-induced obese rats were treated with either S 23521 or vehicle. S 23521 was given either intraperitoneally (10 or 100 microg/kg) or subcutaneously (100 microg/kg) for 14 and 20 days, respectively. Because the low-dose treatment did not affect food intake and body weight, the subcutaneous treatment at high dose was selected to test the effect on selected end-points. RESULTS: Treated obese rats significantly decreased their cumulative energy intake in relation to vehicle-treated counterparts (3401 +/- 65 vs. 3898 +/- 72 kcal/kg per 20 days; p < 0.05). Moreover, their body weight gain was reduced by 110%, adiposity was reduced by 20%, and plasma triglyceride levels were reduced by 38%. The treatment also improved glucose tolerance and insulin sensitivity of obese rats. Regarding gene expression, no changes in uncoupling protein-1, uncoupling protein-3, leptin, resistin, and peroxisome proliferator activated receptor (PPAR)-gamma were observed. DISCUSSION: S 23521 is an effective glucagon-like peptide-1-(7-36) amide analogue, which induced a decrease in energy intake, body weight, and adiposity in a rat model of diet-induced obesity. In addition, the treatment also improved glucose tolerance and insulin sensitivity of obese rats. These results strongly support S 23521 as a putative molecule for the treatment of obesity. PMID- 15536224 TI - Quantitative body composition analysis in awake mice and rats by magnetic resonance relaxometry. AB - OBJECTIVE: Magnetic resonance (MR) relaxometry has recently been introduced for noninvasive body composition analysis in awake mice. The purpose of the present study was to extend the method to rats and to introduce calibration procedures that render MR relaxometry fully quantitative. RESEARCH METHODS AND PROCEDURES: Proton T(2) MR relaxometry at 4.7 Tesla was used for body composition analyses in 700 awake mice and 400 rats of different strains and conditions. Relaxograms calculated from the signal decays observed with multi-spin-echo acquisition provided well-separated contributions of tissue water and fat. Analysis of fat composition was carried out in vivo using (13)C-MR spectroscopy. Evolution of body composition in rats was assessed during drug treatment. RESULTS: MR relaxometry for noninvasive body composition analysis in laboratory rodents was implemented on a standard MR scanner, and a throughput of >30 animals per hour was achieved. Excellent linearity and reproducibility with coefficients of variance as low as 2.5% and 1.7% were obtained in mice and rats, respectively. The lean mass-to-water ratio (mice, 1.35 +/- 0.03; rats, 1.39 +/- 0.04) and the proton density of fat (mice, 8.1 +/- 0.2; rats, 8.9 +/- 0.2 g/mol) were determined from cross-sectional data. Fat composition analysis by (13)C-MR spectroscopy corroborated these findings and yielded information on the average acyl chain length (16.3 +/- 1.6) and contributions of saturated (27 +/- 3%), monounsaturated (22 +/- 2%), and polyunsaturated (51 +/- 3%) fatty acids. Longitudinal assessments in rats treated with sibutramine and dexfenfluramine showed dose-related changes in body composition. DISCUSSION: T(2) MR relaxometry backed by solid calibration provides a powerful means for rapid quantitative body composition analysis in awake mice and rats that is suitable for serial investigations in pharmaceutical research. PMID- 15536225 TI - Adapted changes in left ventricular structure and function in severe uncomplicated obesity. AB - OBJECTIVE: A massive amount of fat tissue, as that observed in obese subjects with BMI over 50 kg/m(2), could affect cardiac morphology and performance, but few data on this issue are available. We sought to evaluate cardiac structure and function in uncomplicated severely obese subjects. RESEARCH METHODS AND PROCEDURES: We studied 55 uncomplicated severely obese patients, 40 women, 15 men, mean age 35.5 +/- 10.2 years, BMI 51.2 +/- 8.8 kg/m(2), range 43 to 81 kg/m(2), with a history of fat excess of at least 10 years, and 55 age-matched normal-weight subjects (40 women, 15 men, mean BMI 23.8 +/- 1.2 kg/m(2)) as a control group. Each subject underwent an echocardiogram to evaluate left ventricular (LV) mass and geometry and systolic and diastolic function. RESULTS: Severely obese subjects showed greater LV mass and indexed LV mass than normal weight subjects (p < 0.01 for all parameters). Nevertheless, LV mass was appropriate for sex, height(2.7), and stroke work in most (77%) uncomplicated severely obese subjects. In addition, no significant difference in LV mass indices and LV mass appropriateness between obese subjects with BMI > or = 50 kg/m(2) and those with BMI < or = 50 kg/m(2) was found. Obese subjects also showed higher ejection fraction and midwall shortening than normal-weight subjects (p = 0.05 and p < 0.01, respectively), suggesting a hyperdynamic systolic function. No significant difference in systolic performance between obese subjects with BMI > or = 50 kg/m(2) and those with BMI < or = 50 kg/m(2) was seen. DISCUSSION: Our data show that uncomplicated severe obesity, despite the massive fat tissue amount, is associated largely with adapted and appropriate changes in cardiac structure and function. PMID- 15536226 TI - Trends in the association between obesity and socioeconomic status in U.S. adults: 1971 to 2000. AB - OBJECTIVE: To study the secular trends in the disparity of obesity across socioeconomic status (SES) groups among U.S. adults. RESEARCH METHODS AND PROCEDURES: We used national representative data collected in the National Health and Nutrition Examination Surveys conducted in 1971 to 1974, 1976 to 1980, 1988 to 1994, and 1999 to 2000 from 28,543 adults 20 to 60 years old. Obesity was defined based on BMI calculated using measured weight and height. Trends in the relationship between obesity and education levels were analyzed controlling for age, gender, and ethnicity. RESULTS: The disparity in obesity across SES (less than high school, high school, and college or above to indicate low, medium, and high SES, respectively) has decreased over the past 3 decades. In National Health and Nutrition Examination Surveys I (1971 to 1974), there was as much as a 50% relative difference in the obesity prevalence across the three groups, but by 1999 to 2000, it decreased to 14%. This trend was more pronounced in women. The trends of diminishing disparities in obesity were also revealed by our logistic and linear regression analyses. The odds ratio converged to 1 from the 1970s to 2000. In most sociodemographic groups, the relationship between BMI and SES (coefficients) has been weakened over time. DISCUSSION: The association between SES and obesity has been weakened over the past 3 decades, when the prevalence of obesity increased dramatically. There are considerable variations in the changes in the associations across gender and ethnic groups. Our findings suggest that individual characteristics are not likely the main cause of the current obesity epidemic in the U.S., whereas social-environmental factors play an important role. Strategies for obesity prevention and management should target all SES groups from a societal perspective. PMID- 15536227 TI - Assessing body composition among 3- to 8-year-old children: anthropometry, BIA, and DXA. AB - OBJECTIVE: To examine the inter-relationships of body composition variables derived from simple anthropometry [BMI and skinfolds (SFs)], bioelectrical impedance analysis (BIA), and dual energy x-ray (DXA) in young children. RESEARCH METHODS AND PROCEDURES: Seventy-five children (41 girls, 34 boys) 3 to 8 years of age were assessed for body composition by the following methods: BMI, SF thickness, BIA, and DXA. DXA served as the criterion measure. Predicted percentage body fat (%BF), fat-free mass (FFM; kilograms), and fat mass (FM; kilograms) were derived from SF equations [Slaughter (SL)1 and SL2, Deurenberg (D) and Dezenberg] and BIA. Indices of truncal fatness were also determined from anthropometry. RESULTS: Repeated measures ANOVA showed significant differences among the methods for %BF, FFM, and FM. All methods, except the D equation (p = 0.08), significantly underestimated measured %BF (p < 0.05). In general, correlations between the BMI and estimated %BF were moderate (r = 0.61 to 0.75). Estimated %BF from the SL2 also showed a high correlation with DXA %BF (r = 0.82). In contrast, estimated %BF derived from SFs showed a low correlation with estimated %BF derived from BIA (r = 0.38); likewise, the correlation between DXA %BF and BIA %BF was low (r = 0.30). Correlations among indicators of truncal fatness ranged from 0.43 to 0.98. DISCUSSION: The results suggest that BIA has limited utility in estimating body composition, whereas BMI and SFs seem to be more useful in estimating body composition during the adiposity rebound. However, all methods significantly underestimated body fatness as determined by DXA, and, overall, the various methods and prediction equations are not interchangeable. PMID- 15536228 TI - Development of a culturally relevant body image instrument among urban African Americans. AB - OBJECTIVE: To validate a culturally relevant body image instrument among urban African Americans through three distinct studies. RESEARCH METHODS AND PROCEDURES: In Study 1, 38 medical practitioners performed content validity tests on the instrument. In Study 2, three research staff rated the body image of 283 African-American public housing residents (75% women, mean age = 44 years), with the residents completing body image, BMI, and percentage body fat measures. In Study 3, 35 African Americans (57% men, mean age = 42) completed body image measures and evaluated their cultural relevance. RESULTS: In Study 1, 97% to 100% of practitioners sorted the jumbled figures into the correct ascending order. The correlation between the body image figures and the practitioners' weight classifications of the figures was high (r = 0.91). In Study 2, observers arrived at similar ratings of body size with excellent consistency (alpha = 0.95). Ratings of body image were strongly correlated with participant BMI (r = 0.89 to 0.93 across observers and 0.81 for all participants) and percentage of body fat (r = 0.77 to 0.89 across observers and 0.76 for all participants). In Study 3, body image ratings with the new scale were positively correlated with other validated figural scales. The majority of participants reported that figures in the new body image scale looked most like themselves and other African Americans and were easiest to identify themselves with. DISCUSSION: The instrument displayed strong psychometric performance and cultural relevance, suggesting that the scale is a promising tool for examining body image and obesity among African Americans. PMID- 15536229 TI - Quantitative genetic analysis of glucose transporter 4 mRNA levels in baboon adipose. AB - OBJECTIVE: Glucose transporter 4 (GLUT4) is an insulin-responsive glucose transporter expressed in adipose tissue. A decrease of the mRNA abundance of GLUT4 in adipose tissue has been observed in conditions of insulin resistance. The objective was to conduct quantitative genetic analyses using GLUT4 mRNA levels in omental adipose tissue of baboons as a novel phenotype. RESEARCH METHODS AND PROCEDURES: A blood sample and a biopsy of omental adipose tissue were collected from 418 adult, pedigreed baboons. Total RNA was isolated from adipose tissue biopsies, and GLUT4 mRNA abundance was assayed by quantitative, real-time reverse transcription-polymerase chain reaction. Insulin and glucose were determined in fasting plasma by standard methods. Quantitative genetic analyses were conducted using GLUT4 mRNA, insulin, and glucose as quantitative traits. RESULTS: GLUT4 mRNA expression in omental adipose tissue was heritable (h(2) = 0.23, p = 0.001). Bivariate genetic analyses revealed a significant genetic correlation (rho(G)) between GLUT4 mRNA abundance and both body weight (rho(G) = 0.63, p = 0.007), BMI (rho(G) = 0.59, p = 0.02) and insulin (rho(G) = 0.72, p = 0.04). A genome scan was conducted, and a quantitative trait locus was detected on chromosome 10p12 with a logarithm of the odds ratio score of 1.1. DISCUSSION: GLUT4 mRNA abundance in omental adipose tissue has a significant genetic component. These findings suggest that expression of GLUT4 mRNA, plasma insulin levels, and body weight may be regulated by common genes. PMID- 15536230 TI - Topiramate: long-term maintenance of weight loss induced by a low-calorie diet in obese subjects. AB - OBJECTIVE: To examine the safety and efficacy of topiramate (TPM) for maintaining weight following a low-calorie diet. RESEARCH METHODS AND PROCEDURES: Obese subjects (30 < or = BMI < 50 kg/m(2)) 18 to 75 years old received a low-calorie diet for 8 weeks. Those who lost > or =8% of their initial weight received TPM (96 or 192 mg/d) or placebo; all were on a lifestyle modification plan. Sixty weeks of medication were planned. Sponsor ended study early to develop a new controlled-release formulation with the potential to enhance tolerability and simplify dosing in this patient population. Efficacy was analyzed in subjects who completed 44 weeks of treatment before study termination. RESULTS: Of the 701 subjects enrolled, 80% lost > or =8% of their initial body weight and were randomized; 293 were analyzed for efficacy. Most withdrawals were due to premature termination of the study. Subjects receiving TPM lost 15.4% (96 mg/d) and 16.5% (192 mg/d) of their enrollment weight by week 44, compared with 8.9% in the placebo group (p < 0.001). Subjects on TPM continued to lose weight after the run-in, whereas those on placebo regained weight. Significantly more TPM subjects lost 5%, 10%, or 15% of their randomization weight than placebo. Most adverse events were related to the central nervous system. DISCUSSION: During a treatment period of 44 weeks, TPM was generally well tolerated, and subjects maintained weight loss initially achieved by a low-calorie diet-and produced additional clinically significant weight loss beyond that achieved by a low-calorie diet. PMID- 15536231 TI - Physical, behavioral, and body image characteristics in a tri-racial group of adolescent girls. AB - OBJECTIVES: The purpose of this study was to evaluate physical characteristics, nutrient intake, physical activity level, and body image in white (CC), African American (AA), and Hispanic-American (HA) female adolescents. RESEARCH METHODS AND PROCEDURES: High school volunteers were solicited for this study. Self reported information was used to determine subject characteristics, family income, physical activity, body image, and nutrient intake. Physical evaluations were used to determine body mass index, percent body fat, fat distribution, resting heart rate, and blood pressure (BP). RESULTS: Results showed that AA girls displayed significantly higher diastolic BP than HA girls (p = 0.029). CC adolescents showed greater physical activity (p = 0.010) and lower adiposity than HA adolescents (p = 0.048), as well as lower subscapular skinfold than AA adolescents (p = 0.018). AA adolescents selected a higher ideal body size than CC girls (p = 0.038). There was also a significant difference in percentage carbohydrates (p < 0.034) and cholesterol consumed (p < 0.016) among groups, with CC girls showing the highest values for carbohydrates and lowest values for cholesterol intake among groups. DISCUSSION: Given our findings of higher adiposity and lower physical activity levels in HA adolescents and greater diastolic BP levels and subscapular skinfold in AA adolescents, more interventions should be targeted toward improving health-related variables among minority populations. PMID- 15536232 TI - Obesity and awareness of obesity as risk factors for breast cancer in six ethnic groups. AB - OBJECTIVE: To document BMI and knowledge regarding obesity as a risk factor for breast cancer among subpopulations of African-, Caribbean-, and European-American women and to consider the variables predicting obesity in these diverse groups. RESEARCH METHODS AND PROCEDURES: A stratified cluster-sampling plan was used to recruit 1364 older women from Brooklyn, NY, during 2000-2002. Two groups were born in the United States (African Americans and European Americans), whereas others were from the English-speaking Caribbean, Haiti, the Dominican Republic, and Eastern Europe. Participants provided demographics, height and weight measures, and estimates of the risk obesity posed for breast cancer. RESULTS: Women from all groups were significantly overweight (BMI > 25 kg/m(2)), although European Americans were lowest, followed by Dominicans and Haitians; African American and English-speaking Caribbean women fell into the obese range, even when background variables were controlled. Knowledge of obesity as a breast cancer risk factor was also poor across groups, but Dominicans and Haitians had the lowest scores on knowledge. Importantly, knowledge was not associated with BMI in the overall sample, even when controlling for demographics and ethnicity, although logistic regressions comparing normal weight women with overweight and obese groupings suggested some knowledge of breast cancer risk in the overweight, but not the obese, group. DISCUSSION: The findings remind health professionals of the need to consider more specific ethnic groupings than has hitherto been the case, as well as consider how ethnic and cultural variables may influence perceptions of obesity and its relation to cancer risk. PMID- 15536233 TI - Ucp3 expression during weight gain and loss, cold exposure, and fasting in the collared lemming. AB - OBJECTIVE: To determine the gene sequence and tissue distribution of uncoupling protein 3 (Ucp3) in the collared lemming, we quantified mRNA expression of Ucp3 under known states of altered energy expenditure (photoperiod-induced weight gain and loss, cold exposure, and fasting) and measured mitochondrial oxygen consumption to assess possible functional changes in energy expenditure. RESEARCH METHODS AND PROCEDURES: The Ucp3 gene sequence information was obtained using the reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends methods. Northern blots were used to determine mRNA expression levels. Respirometry was used to measure oxygen consumption rates in isolated mitochondria. RESULTS: The lemming Ucp3 gene has a 97% sequence similarity with other published Ucp3 sequences at the amino acid level. Ucp3 mRNA is expressed in muscle, heart, and brown adipose tissue of collared lemmings. Long-photoperiod lemmings have a higher expression of Ucp3 mRNA than short-photoperiod lemmings (p < 0.001) in both muscle and brown adipose tissue. Transferring lemmings from long to short photoperiods (inducing weight gain) significantly decreased Ucp3 mRNA expression (p < 0.01), whereas transferring lemmings from short to long photoperiods (inducing weight loss) significantly increased Ucp3 expression (p < 0.001). Muscle Ucp3 mRNA expression was significantly decreased by 10 days of mild (10 degrees C) cold exposure (p < 0.001). Muscle Ucp3 mRNA expression was significantly increased by fasting (p < 0.01) and was correlated to plasma free fatty acid levels (r = 0.7). Photoperiod transfer did not alter mitochondrial coupling. DISCUSSION: These data suggest that UCP3 may not be involved in energy expenditure in the collared lemming. PMID- 15536234 TI - Preventing overestimation of pixels in computed tomography assessment of visceral fat. AB - OBJECTIVE: Computed tomography (CT) is a common research procedure for measuring abdominal fat distribution, but little is written about the software used to analyze images. Our objective was to compare in-house and commercially available software for quantitative measurement of abdominal fat distribution. In the process, we encountered some unexpected problems. RESEARCH METHODS AND PROCEDURES: A total of 123 volunteers had single-slice abdominal CT images taken that were used to evaluate various aspects of the commercial image analysis program. RESULTS: The agreement between the commercial and in-house programs was excellent (r = 0.996, p < 0.00,001) for both total and intraabdominal fat, and we were able to reduce between-observer variability in measured fat areas through the use of statistical handling of region of interest information. We also noted that intracolonic contents sometimes had the same Hounsfield units as adipose tissue. We analyzed single-slice CT images from 50 volunteers to determine the potential impact of this effect on visceral fat area; the overestimate of visceral fat area was 19 +/- 22% (maximum, 112% overestimate). The commercial program could prevent this error, whereas our in-house program could not. DISCUSSION: We concluded that a readily available commercial image analysis program compares well with a previously validated in-house program and that it offers some advantages with respect to preventing overestimation of pixels as visceral fat. PMID- 15536235 TI - Are sexual dissatisfaction and sexual abuse associated with obesity? A population based study. AB - OBJECTIVE: To investigate whether there is any association between obesity and sexual satisfaction and sexual abuse in a normal population. RESEARCH METHODS AND PROCEDURES: A representative sample of 2810 subjects from a population study was interviewed about sexual satisfaction, sexual abuse, and life satisfaction. The answers from normal weight, overweight, and obese participants were compared. Univariate and multivariate analyses were performed. RESULTS: Data were presented separately for two age groups, 18 to 49 and 50 to 74 years, and gender. The older group of obese men reported a greater decrease of sexual desire compared with 5 years prior than normal weight men [odds ratios (OR), 2.44; 95% confidence interval (CI), 1.4 to 4.3]. The older group of overweight men reported involuntary participation in sexual activities more often than normal weight men (OR, 2.06; 95% CI, 1.1 to 3.8). Although older overweight and obese women were diagnosed with a lingering disease (defined as >1 month) more often than normal weight women (overweight: OR, 2.41; 95% CI, 1.3 to 4.4; obese: OR, 4.45; 95% CI, 1.7 to 11.5), there was no difference between BMI groups in satisfaction with physical health. DISCUSSION: Overweight and obese groups seem to be heterogeneous with respect to sexual satisfaction and experiences of sexual abuse. No significant differences were detected between BMI groups, which does not exclude the possibility of significant differences between BMI groups among patients seeking medical attention. PMID- 15536236 TI - Interference and facilitation among personal goals: differential associations with subjective well-being and persistent goal pursuit. AB - Three studies demonstrate that mutual facilitation and interference among personal goals are distinct characteristics rather than mutually exclusive opposites and have different functions for psychological well-being and goal pursuit. The three studies vary in design (cross-sectional, short termlongitudinal) and follow a multimethod approach using questionnaires, diaries, and objective behavioral information. Results show that interference among goals (resulting from resource constraints and incompatible goal attainment strategies) is negatively associated with trait and state well-being, whereas mutual facilitation among goals (resulting from instrumental goal relations and overlapping goal attainment strategies) is positively associated with involvement in goal pursuit. PMID- 15536237 TI - Ageism and death: effects of mortality salience and perceived similarity to elders on reactions to elderly people. AB - The present research investigated the hypotheses that elderly people can be reminders of our mortality and that concerns about our own mortality can therefore instigate ageism. In Study 1, college-age participants who saw photos of two elderly people subsequently showed more death accessibility than participants who saw photos of only younger people. In Study 2, making mortality salient for participants increased distancing from the average elderly person and decreased perceptions that the average elderly person possesses favorable attitudes. Mortality salience did not affect ratings of teenagers. In Study 3, these mortality salience effects were moderated by prior reported similarity to elderly people. Distancing from, and derogation of, elderly people after mortality salience occurred only in participants who, weeks before the study, rated their personalities as relatively similar to the average elderly person's. Discussion addresses distinguishing ageism from other forms of prejudice, as well as possibilities for reducing ageism. PMID- 15536239 TI - Every cloud has a silver lining: interpersonal and individual differences determinants of anger-related behaviors. AB - Two studies examined the effect of status and liking of the anger target on anger behavior and individual differences in anger-related behavior. Participants recalled anger instances in which the anger target was of higher/equal/lower status and/or liked/ unfamiliar/disliked; subsequently, they indicated which behaviors they had displayed. In both studies, anger behaviors could be grouped into behaviors that imply approaching the target (anger-out, assertion, reconciliation) and behaviors that reflect avoidance/anger-in or social sharing. The results demonstrated that approach behaviors more likely occur toward lower status or liked targets; avoidance behaviors and social sharing more likely occur when the target is of higher status or disliked. On an individual differences level, an approach and an avoid/social sharing person class were identified. The findings suggest that anger may motivate prosocial behavior or social sharing, depending on the individual and type of relation with the target. Only few gender differences were found. PMID- 15536238 TI - Perspective and prejudice: antecedents and mediating mechanisms. AB - The present work investigated mechanisms by which Whites' prejudice toward Blacks can be reduced (Study 1) and explored how creating a common ingroup identity can reduce prejudice by promoting these processes (Study 2). In Study 1, White participants who viewed a videotape depicting examples of racial discrimination and who imagined the victim's feelings showed greater decreases in prejudice toward Blacks than did those in the objective and no instruction conditions. Among the potential mediating affective and cognitive variables examined, reductions in prejudice were mediated primarily by feelings associated with perceived injustice. In Study 2, an intervention designed to increase perceptions of a common group identity before viewing the videotape, reading that a terrorist threat was directed at all Americans versus directed just at White Americans, also reduced prejudice toward Blacks through increases in feelings of injustice. PMID- 15536240 TI - The role of defensive confidence in preference for proattitudinal information: how believing that one is strong can sometimes be a defensive weakness. AB - This series of studies identified individuals who chronically believe that they can successfully defend their attitudes from external attack and investigated the consequences of this individual difference for selective exposure to attitude incongruent information and, ultimately, attitude change. Studies 1 and 2 validated a measure of defensive confidence as an individual difference that is unidimensional, distinct from other personality measures, reliable over a 2-week interval, and organized as a trait that generalizes across various personal and social issues. Studies 3 and 4 provided evidence that defensive confidence decreases preference for proattitudinal information, therefore inducing greater reception of counterattitudinal materials. Study 5 demonstrated that people who are high in defensive confidence are more likely to change their attitudes as a result of exposure to counterattitudinal information and examined the perceptions that mediate this important phenomenon. PMID- 15536241 TI - When what you know is not enough: expertise and gender dynamics in task groups. AB - This study investigates how the contribution, identification, and consideration of expertise within groups are affected by gender differences. The authors examined the effects of member expertise and gender on others' perceptions of expertise, actual and own perceptions of influence, and group performance on a decision-making task. The authors' findings are consistent with social role theory and expectation states theory. Women were less influential when they possessed expertise, and having expertise decreased how expert others perceived them to be. Conversely, having expertise was relatively positive for men. These differences were reflected in group performance, as groups with a female expert underperformed groups with a male expert. Thus, contrary to common expectations, possessing expertise did not ameliorate the gender effects often seen in workgroups. The findings are discussed in light of their implications for organizational workgroups in which contribution of expertise is critical to group performance. PMID- 15536242 TI - Gender, social class, and the subjective experience of aging: self-perceived personality change from early adulthood to late midlife. AB - This study explored the applicability of previous research (obtained with groups of college-educated women) about the subjective experience of aging in midlife to men and less-educated people. Two-hundred fifty-nine men and women who graduated from a public high school in 1955-1957 retrospectively assessed their feelings of identity certainty, confident power, generativity, and concern about aging for their 60s, 40s, and 20s. Participants reported higher levels of identity certainty, confident power, and concern about aging at each age, and a leveling off of generativity in their 60s. There were some gender and social class differences. Although men and women recalled the same trajectory of these feelings, men reported higher levels of identity certainty and confident power across age. Non-college-educated men recalled the highest levels of concern about aging across age. We discuss how these findings add to our understanding of the experience of aging in these domains. PMID- 15536243 TI - A threat in the computer: the race implicit association test as a stereotype threat experience. AB - Three experiments test whether the threat of appearing racist leads White participants to perform worse on the race Implicit Association Test (IAT) and whether self-affirmation can protect from this threat. Experiments 1 and 2 suggest that White participants show a stereotype threat effect when completing the race IAT, leading to stronger pro-White scores when the test is believed to be diagnostic of racism. This effect increases for domain-identified (highly motivated to control prejudice) participants (Experiment 2). In Experiment 3, self-affirmation inoculates participants against stereotype threat while taking the race IAT. These findings have methodological implications for use of the race IAT and theoretical implications concerning the malleability of automatic prejudice and the potential interpersonal effects of the fear of appearing racist. PMID- 15536244 TI - Cognitions associated with attempts to empathize: how do we imagine the perspective of another? AB - Although the theoretical importance of perspective taking has long been recognized, surprisingly little work has documented the cognitions associated with attempts to imagine another's point of view. To explore this issue and to determine whether perspective taking increases the likelihood of self-related thoughts, two experiments were carried out. In the first, a thought-listing procedure was used to assess observer cognitions; in the second, a less reactive measure was used. Instructions to imagine the self in the target's position and instructions simply to imagine the target's perspective produced increased levels of self-related cognition relative to a traditional control condition; the imagine-self condition also produced more self-thoughts and fewer target thoughts than did the imagine-target condition. The control condition produced thoughts suggesting that the observers were distancing themselves from the target. Observers receiving no instructions at all reported cognitions that closely resembled those of observers who received imagine-target instructions. PMID- 15536245 TI - Mea culpa: predicting stock prices from organizational attributions. AB - People's causal attributions for events in their lives have been shown to relate to individual and interpersonal outcomes. Groups and organizations also make causal attributions, and this article examines whether their publicly communicated attributions predict organizational-level outcomes. By content analyzing attributions contained in corporate annual reports from 14 companies during a 21-year period, the authors found that organizations that made "self disserving" attributions- internal and controllable attributions for negative events-had higher stock prices 1 year later. The authors argue that claiming personal responsibility for negative events made the organizations appear more in control, leading to more positive impressions. PMID- 15536246 TI - Do normative transgressions affect punitive judgments? An empirical test of the psychoanalytic scapegoat hypothesis. AB - According to psychoanalytic theory, punitiveness is based on a projection of one's own immoral desires and the moral conflict they cause (scapegoat hypothesis). This hypothesis implies that transgressors impose harsher punishment onto comparable wrongdoers. This effect should be amplified by strength of decision conflict. An alternative hypothesis based on blameavoidance motivation is derived. Participants (N = 291) were asked to indicate whether they would commit an unlawful act in a moral temptation situation and how conflicted they felt in making their decision. Later, they had to judge convicts in criminal cases that were similar to the previous temptation situations. Authoritarianism was assessed as covariate. In contrast to the scapegoat but consistent with the blame-avoidance interpretation, transgressors were more lenient than nontransgressors. Authoritarianism had main effects on punitiveness. Decision conflict was neither directly nor indirectly related to punitiveness. The findings challenge the validity of the scapegoat hypothesis. PMID- 15536247 TI - Essentialist beliefs about personality and their implications. AB - Two studies examine implicit theories about the nature of personality characteristics, asking whether they are understood as underlying essences. Consistent with the hypothesis, essentialist beliefs about personality formed a coherent and replicable set. Personality characteristics differed systematically in the extent to which they were judged to be discrete, biologically based, immutable, informative, consistent across situations, and deeply inherent within the person. In Study 1, the extent to which characteristics were essentialized was positively associated with their perceived desirability, prevalence, and emotionality. In Study 2, essentialized characteristics were judged to be particularly important for defining people's identity, for forming impressions of people, and for communicating about a third person. The findings indicate that people understand some personality attributes in an essentialist fashion, that these attributes are taken to be valued elements of a shared human nature, and that they are particularly central to social identity and judgment. PMID- 15536250 TI - Genetically engineered monogamy in voles lends credence to the modus operandi of behavioural ecology. PMID- 15536251 TI - Benefits of being biased! PMID- 15536252 TI - Chance in our strands? PMID- 15536253 TI - Sex determining signal in Drosophila melanogaster. PMID- 15536254 TI - The distribution of sex-factors in the X-chromosome of Drosophila melanogaster. 1934. PMID- 15536255 TI - REML estimates of genetic parameters of sexual dimorphism for wing and thorax length in Drosophila melanogaster. AB - Restricted maximum likelihood was used to estimate genetic parameters of male and female wing and thorax length in isofemale lines of Drosophila melanogaster, and results compared to estimates obtained earlier with the classical analysis of variance approach. As parents within an isofemale line were unknown, a total of 500 parental pedigrees were simulated and mean estimates computed. Full and half sibs were distinguished, in contrast to usual isofemale studies in which animals were all treated as half sibs and hence heritability was overestimated. Heritability was thus estimated at 0.33, 0.38, 0.30 and 0.33 for male and female wing length and male and female thorax length, respectively, whereas corresponding estimates obtained using analysis of variance were 0.46, 0.54, 0.35 and 0.38. Genetic correlations between male and female traits were 0.85 and 0.62 for wing and thorax length, respectively. Sexual dimorphism and the ratio of female to male traits were moderately heritable (0.30 and 0.23 for wing length, 0.38 and 0.23 for thorax length). Both were moderately and positively correlated with female traits, and weakly and negatively correlated with male traits. Such heritabilities confirmed that sexual dimorphism might be a fast evolving trait in Drosophila. PMID- 15536256 TI - Genetic and genotype x environment interaction effects for the content of seven essential amino acids in indica rice. AB - It is necessary for rice breeders to understand the genetic basis of nutrient quality traits of rice. Essential amino acids are most important in determining the nutrient quality of rice grain and can affect the health of people who depend on rice as a staple food. In view of the paucity of genetic information available on essential amino acids in indica rice, we estimated the genetic main effects and genotype x environment (G x E) interaction effects on the content of essential amino acids. Nine cytoplasmic male sterile lines as females and five restorer lines as males were introduced in a North Carolina II design across environments. Estimates of the content of the essential amino acids valine, methionine, leucine and phenylalanine showed that they were mainly controlled by genetic main effects, while the contents of threonine, cysteine and isoleucine were mainly affected by G x E effects. In the case of genetic main effects, both cytoplasmic and maternal genetic effects were predominant for all essential amino acids, indicating that selection for improving essential amino acid content based on maternal performance would be more effective than that based on seeds. The total narrow-sense heritabilities were high and ranged from 0.72 to 0.83. Since general heritabilities for these essential amino acids (except for cysteine) were found to be much larger than G x E interaction heritability, the improvement of content of most essential amino acids under selection would be expected under various environments. Rice varieties such as Zhenan 3, Yinchao 1, T49, 26715, 102 and 1391 should be selected as optimal parents for increasing the content of most essential amino acids, while the total genetic effects from Zhexie 2, Xieqingzao, Gangchao 1, V20, Zuo 5 and Zhenshan 97 were mainly negative and these parents could decrease the contents of most essential amino acids. PMID- 15536257 TI - Variation in the relative magnitude of intraspecific and interspecific competitive effects in novel versus familiar environments in two Drosophila species. AB - Models of competitor coevolution, especially the genetic feedback hypothesis, suggest that a negative correlation between intraspecific and interspecific competitive effects may be important in sustaining competitor coexistence, and can give rise to oscillatory dynamics with repeated reversals of competitive superiority. I reanalyzed previously published census data from an experiment in which populations of Drosophila melanogaster and D. simulans underwent competitive coevolution in one familiar and two novel environments, to specifically look for any evidence of a negative relationship between intraspecific and interspecific competitive effects on population growth rates, and for any indication of short period cycling in the relative magnitude of intraspecific and interspecific competitive effects. While there was considerable variation in the relative magnitude of intraspecific and interspecific competitive effects over generations, among both populations and environments, there was no clear evidence supporting the genetic feedback hypothesis. Intraspecific and interspecific competitive effects on population growth rates were strongly positively correlated in novel environments, and uncorrelated in the familiar environment. Data from the familiar environment indicated that indices of competition of populations of the initially superior competitor, D. melanogaster, might be showing some cyclic behaviour, but I argue that this is likely to be transient, and not suggestive of sustained oscillatory dynamics predicted by the genetic feedback model. I discuss the results in the context of the importance of the genetic architecture of intraspecific and interspecific competitive abilities in determining the coevolutionary trajectory of competitive interactions. PMID- 15536259 TI - PCR-based detection of allergenic mackerel ingredients in seafood. PMID- 15536258 TI - Cytochrome P450c17alpha 5'-untranslated region *T/C polymorphism in endometriosis. AB - Estrogen plays a role in the pathogenesis of endometriosis. The CYP17 gene codes for the cytochrome P450c17alpha enzyme that is involved in the estrogen biosynthesis. We aimed to investigate if CYP17 polymorphism could be used as marker to predict the susceptibility of endometriosis. Women were divided into two groups: (1) severe endometriosis (n=119); (2) non-endometriosis groups (n=128). A 169-bp fragment encompassing the T/C polymorphic site in 5' untranslated promoter region (5'-UTR) of the CYP17 was amplified by the polymerase chain reaction, treated with restriction enzyme MspA1I, and electrophoresis. The polymorphism was divided into restriction-enzyme indigestible (T homozygote), T/C heterozygote, and digestible (C homozygote). Genotypes and allelic frequencies for this polymorphism in both groups were compared. We observed a higher but non-significant percentage of T homozygote in the endometriosis women compared with the non-endometriosis women. Proportions of T homozygote / heterozygote / C homozygote for CYP17 in both groups were: (1) 26.1/46.2/27.7% and (2) 17.2/45.3/37.5% (p-value=0.131). T allele was related with higher susceptibility of endometriosis. T and C allele frequencies in both groups were: (1) 49.2/50.8%; (2) 39.8/60.2% (p-value=0.046). Despite the CYP17* T allele appearing to be associated with a trend of increased risk of endometriosis, CYP17 5'-UTR gene polymorphism might not be a useful marker for prediction of endometriosis susceptibility. PMID- 15536260 TI - Short tandem repeat (STR) polymorphisms in Turkish population. PMID- 15536261 TI - From fruit flies to fallout: Ed Lewis and his science. PMID- 15536264 TI - Introductory keynote. The state of the art in animal experimentation. AB - This issue of Annali dell'Istituto Superiore di Sanita critically evaluates the progression in the ethical relationship between humans and animals (i.e., between biomedical researchers and laboratory animals). The main goal of this publication is in fact to provide a basis for an in-depth discussion of highly debated topics in the contemporary bioethics of animal experimentation, with particular focus on ethological issues and the question of minimal sample size and its contribution to reducing the number of experimental animals. The ethological issues addressed in this publication cover all behavioural patterns specifically moulded by the phylogenetic and ontogenetical history of a given species, a perspective that is often neglected when discussing the promotion of animal welfare. PMID- 15536265 TI - Biostatistical approaches to reducing the number of animals used in biomedical research. AB - For ethical reasons, the least number of animals possible should be used in biomedical research, though not so few as to fail to detect biologically important effects or to necessitate the repetition of experiments. We describe biostatistical approaches that can contribute to either reducing the number of animals in single experiments or to increasing the quality of studies so that fewer subsequent studies (and thus animals) will be needed. The described approaches regard different phases of experimentation, specifically: planning the experimental design and calculating the sample size, controlling variability, choosing the response variable, postulating the statistical hypothesis to be tested, choosing the procedure for analysing data, and interpreting and suitably presenting the results. PMID- 15536266 TI - Detecting assumption violations in mixed-model analysis of variance. AB - Parametric analysis of variance (ANOVA) is frequently used to analyse experimental data, yet for the results to be considered as accurate, certain assumptions must be respected: the normality of the distribution of the sampled data, the homogeneity of variance among the groups being compared (i.e., homoscedasticity), and, in certain cases, sphericity. The present work focuses on the methods for detecting violations of these assumptions and provides an example of the application of these methods. PMID- 15536267 TI - Accounting for historical information in designing experiments: the Bayesian approach. AB - Two of the most important statistical problems in human and animal experimentation are the selection of an appropriate number of units to include in a given study and the allocation of these units to the various treatments. Properly addressing these issues allows the units to be used as efficiently as possible, which can contribute to addressing the overall issue of reducing the number of subjects in experimentation. To do so, reliable historical information is of particular importance. In the present paper, we describe the Bayesian approach to determining sample size and allocating units, with particular regard to how the use of historical data can be optimised. The paper focuses on two specific problems: the comparison of normal means and of binomial proportions. PMID- 15536268 TI - Environmental enrichment techniques in non-human primates. The case of Callitrichids. AB - In this paper, we discuss issues concerning the welfare of non-human primates used in laboratory research from an eco-ethological standpoint and suggest means of improving welfare. Following a brief review of the use of non-human primates in European countries and of the legislation that governs this use, we illustrate how a thorough eco-ethological knowledge of the species being studied can play a vital role in improving both its conditions and the quality of the experimental protocols, arguing that the animal's quality of life is closely linked to the quality of data. As a model for describing environmental enrichment techniques, we have used the common marmoset (Callithrix jacchus). PMID- 15536269 TI - Health-promoting factors and animal welfare. AB - The notion of "health" as the absence of disease has been replaced by a more holistic view based on the concept of "total wellbeing" and its interrelated physiological, psychological, and social components. This approach has radically changed biomedical research, both in terms of experimental methodology and in addressing ethical issues such as animal welfare. The present work focuses on the entire range of factors that can affect the wellbeing of experimental animals, including the physical, social, and psychological conditions of the housing environment, and the animal's behavioural and biological reaction to this environment. PMID- 15536270 TI - Ethological and methodological considerations in the use of newborn rodents in biomedical research. AB - The use of newborn or immature animals in biomedical research poses certain challenges, in that the ecological niche of immature animals differs from that of adults, and the stimuli provided to immature animals can have profound physiological and behavioural effects through adulthood, even at the cerebral level. In particular, the newborn's behaviour and physiology are regulated by olfactory, thermal, and tactile stimulation supplied by the mother in the nest environment. Thus any disturbance to the mother-offspring relationship in the first two weeks of life can profoundly change the physiological and psychological state of the young. For this reason, research experiments must be adapted to the specific characteristics of the immature animal, to improve not only the quality of the data obtained but also the welfare of the animals. To this end, the present work provides some ethological and methodological considerations in the maintenance, handling, and testing of newborn rats and mice in biomedical research. PMID- 15536271 TI - Rules of good practice in the care of laboratory animals used in biomedical research. AB - In recent years, the use of laboratory animals has decreased as a result of the adoption of alternative methods such as in vitro experiments and simulation studies. Nonetheless, animal models continue to be necessary in many fields of biomedical research, giving rise to ethical issues regarding the treatment of these animals. In the present work, a general overview of the rules of good practise in caring for laboratory animals is provided, focussing on housing conditions and the proper means of handling animals, including the importance of the relationship or "bond" between the researcher and the animal. PMID- 15536272 TI - Animal experimentation in Italy. Legislation and the authorization of research protocols. AB - In Italy, the European Directive 86/609/EEC ("The Council Directive on the approximation of laws, regulations and administrative provisions of the Member States regarding the protection of animals used for experimental and other scientific purposes") has been given effect with Legislative Decree 116/92, whose main objective is to guarantee the welfare of animals used in research and to prevent unnecessary experiments on animals from being carried out. The regulatory authority for controlling the use of laboratory animals in Italy is the Ministry of Health, which requires that researchers requesting authorization to perform experiments on animals submit not only a copy of the experimental protocol but also a detailed application form that focuses on how the animals will be used. In the evaluation process, the Istituto Superiore di Sanita (Italy's National Institute of Health), in particular, the Service for Biotechnology and Animal Welfare, plays a key decision-making role. The evaluation is conducted by experts in the given area of research and by a veterinarian specifically trained in evaluating experimental protocols involving animal use. In the present work, the evaluation process is explained and a point-by-point description of the application form is provided. PMID- 15536273 TI - Non-invasive techniques for analysing hormonal indicators of stress. AB - The growing concern over animal welfare has led to an increased awareness of the need to monitor and reduce stress in laboratory, zoo, and farm animals. To do so, valid and reliable methods are necessary. In the present work, we discuss non invasive techniques for analysing hormonal indicators of stress, particularly glucocorticoids. Specifically, we describe methods for analysing samples of saliva, urine, and faeces, the collection of which, unlike blood sampling, does not represent a source of stress and thus a potential source of bias. We also address species-specific responses to stress and inter-individual variability, the reliability of specific hormonal assays, and the use of indirect measures of circulating stress hormones. PMID- 15536274 TI - Refinement of pain evaluation techniques. The formalin test. AB - The formalin test is an important means of assessing pain evoked-behaviours induced by moderately intense and long-lasting noxious stimuli, yet the procedures for performing the test have not been standardised, hindering comparisons of data. In rodents, the level of sensitivity to the test and the evoked response have been observed to change depending on a number of variables. Among them, gender is the most important, although many other variables can influence the behavioural response (e.g., the animal species, the strain of mice and rats, housing conditions, and early social experience). In the present work, we provide information on the procedures used to perform the formalin test and the effects of both these procedures and the conditions and manipulation prior to the test on the animal and its behaviour, with the ultimate goal of improving these procedures and conditions and decreasing the variability among laboratories. PMID- 15536275 TI - Refining learning and memory assessment in laboratory rodents. An ethological perspective. AB - In neuroscientific research, the importance of a multi-level approach in studying behaviour, ranging from the molecular to the behavioural level, has been increasingly recognised. In fact, behaviour represents the ultimate output of the brain, and behavioural phenotyping may provide functional information that may not be detectable using molecular, cellular, or histological evaluations. To correctly exploit the study of behaviour in the field of neuroscience, the principles and strategies of ethology must be carefully considered, so as to design appropriate experiments and accurately interpret data. In particular, ethological-type scoring, which entails both the close observation of the studied behaviour and the taking into account of its possible evolutionary history and adaptive significance, can contribute to standardising experimental protocols, so as to improve the quality of data and the welfare of the experimental animal. PMID- 15536276 TI - Refinement techniques in experimental protocols involving Callitrichids. AB - The invasiveness of biomedical experiments on laboratory animals should be limited to the greatest extent possible yet without sacrificing the quality of the data collected. To this end, refinement techniques can be used. In the present work, we describe some of these techniques, focussing on the familiarity of the experimental environment, alternative sampling techniques (including the use of positive training), telemetry, and methods for improving ethological experiments. As a model, we have chosen the common marmoset (Callithrix jacchus), which is frequently used in biomedical research. PMID- 15536277 TI - Metabolic and cardiovascular disorders in highly inbred lines for intensive pig farming: how animal welfare evaluation could improve the basic knowledge of human obesity. AB - Intensive pig farming is currently performed with highly inbred lines of animals characterised not only by fast growing and low proportion of back-fat tissue, but also by an impaired development of muscular mass and cardiocirculatory system, and higher susceptibility to oxidative stress. This picture of genetically determined metabolic alterations suggests the use of these lines as a nice tool for conducting comparative studies with human populations affected by high incidence of obesity and their sequelae, e.g. type 2 diabetes, arteriosclerosis, and cardiovascular diseases. The animal model offers, in fact, the advantages of low genetic variance, homogeneous feeding regime, and the absence of confounding factors typical of humans, such as smoking, alcohol drinking, etc. The similarity of pathological response to high caloric intakes between the two species supports the use of pig model for identifying genes and their variants associated with energy storage defects through the activation of both hormonal and biochemical pathways. To this purpose, the evaluation of animal welfare constitutes a nice opportunity to deepen the matter, through the evaluation of both ethical and physiological needs. PMID- 15536278 TI - [An experience with health education for high school students]. AB - In Italy, measures for the prevention of drug addiction have often been directed solely towards providing information and creating greater awareness. Fisher and Fisher (1992), instead, propose a model based on three factors, Information Motivation-Behavioural Skills, for the planning of programs for the prevention of health risk behaviour. This model provided the basis for the pilot experience of a health education course for the prevention of drug use, which involved 417 students in two high schools in Rome. This experience shows that efforts to prevent drug use in young people, if focused only on content, risk failure because young people already seem to possess the basic information concerning the risks connected to substance abuse. For this reason, there is a need for the organization of encounters for confrontation and debate in order to favour the acquisition of the capabilities needed to confront a potential problem of substance abuse. PMID- 15536280 TI - Risk assessment of external events in nuclear facilities. AB - After the Chernobyl accident, a National Emergency Plan of protective measures for radiological emergencies has been set up in Italy to cope with nuclear risks which require actions at national level. Since the Italian nuclear power plants are, at present, not operational, the most relevant nuclear risk source identified in the National Emergency Plan is related to an accident occurring in a nuclear power plant near the Italian borders. However, risks related to severe accidents to other nuclear facilities present in Italy, such as provisional radioactive waste deposits or research centers are not taken into account in the Plan. In this paper the hypothetical radiological impact of a severe external event in a spent fuel storage pool has been evaluated, as this event appears to be one of those with the most severe consequences. PMID- 15536279 TI - [Pleural mesothelioma and environmental exposure to mineral fibres: the case of a rural area in the Basilicata region, Italy]. AB - The main aspects of the sites characterized by environmental exposure to mineral asbestiform fibres are described. Several adverse health effects including high incidence of pleural mesothelioma are reported. The average concentration of airborne fibres is generally low but it rises significantly in association with mechanical disturb of materials with fibres. Multiple sources of exposure have been identified, fibres can be found in the soil and in many materials locally used, mainly in buildings. Three mesothelioma cases were observed in a small rural area of the Basilicata region (Italy). Two of them had a possible occupational exposure to asbestos, the third had a proved environmental exposure to tremolite. This fibre, found in the area, is the same observed in two of the three biological samples analysed. PMID- 15536281 TI - Role of incretins in pancreas growth and development. PMID- 15536282 TI - The functional angiotensin converting enzyme gene I/D polymorphism does not alter susceptibility to chronic pancreatitis. AB - CONTEXT: Alterations of the renin-angiotensin system have been implicated in the pathogenesis of various diseases. The angiotensin converting enzyme is a key enzyme in the renin-angiotensin system. A deletion polymorphism of a 287-bp fragment of intron 16 of the angiotensin converting enzyme gene allele results in higher levels of circulating enzyme. ACE deletion genotype has been linked to heart diseases, sarcoidosis and liver fibrosis. The pancreatic renin-angiotensin system plays a role in the development of pancreatic fibrosis and ACE inhibitors decrease pancreatic fibrosis in experimental models. OBJECTIVES: We investigated the frequency of the ACE gene insertion/deletion polymorphism in chronic pancreatitis patients and controls. PATIENTS: Subjects with familial pancreatitis (n=51), sporadic chronic pancreatitis (n=104), and healthy controls (n=163) were evaluated. MAIN OUTCOME MEASURE: The presence of ACE insertion/deletion polymorphism. RESULTS: The frequency of the ACE gene deletion allele was similar in familial pancreatitis (49.0%) sporadic pancreatitis (51.0%) and controls (55.8%). Furthermore, there was no significant difference in clinical features between patients with ACE-insertion or insertion/deletion genotypes vs. patients with ACE-deletion genotype. CONCLUSION: We conclude that the ACE deletion genotype does not make a significant contribution to the pathogenesis and the progression of chronic pancreatitis. PMID- 15536283 TI - Cholecystokinin octapeptide: a potential growth factor for pancreatic beta cells in diabetic rats. AB - CONTEXT: Diabetes is associated with the reduction of beta cell mass and activity. Cholecystokinin (CCK) is known to induce growth of the exocrine pancreas and to stimulate insulin secretion. OBJECTIVE: We investigated the possible role of CCK-octapeptide (CCK-8) in generating islet cell proliferation in type 1 and type 2 diabetic rats. METHODS: Streptozotocin-induced type 1 diabetic rats, streptozotocin/nicotinamide-induced type 2 diabetic rats and non diabetic rats were subjected to CCK-8 (1, 2 and 4 microg/kg) or saline injections (for the control group), three times daily for 8 successive days. MAIN OUTCOME MEASURES: The islets of Langerhans were analyzed morphometrically; the beta-cell function was evaluated by an oral glucose tolerance test, and plasma basal glucose and insulin concentrations. RESULTS: In type 1 diabetic rats, CCK-8 induced an increase in beta cell surface associated with a marked increase in the mitotic index; this effect appeared at a concentration of 1 microg/kg CCK-8 and was the highest at a concentration of 4 microg/kg CCK-8. In addition, pancreatic- and plasma-insulin concentrations increased while fasting blood glucose concentrations were reduced when compared to saline-treated rats but the glycemic response to an oral glucose challenge did not significantly improve. In type 2 diabetic rats and in non-diabetic rats, CCK-8 treatment did not significantly affect either the structure or the functional state of beta-cells. CONCLUSIONS: CCK-8 could improve blood glucose concentrations in type 1 diabetic rats correlated with an increase in beta cell mass probably potentiated by the chronic hyperglycemic state. PMID- 15536284 TI - Intussusception caused by a heterotopic pancreas. Case report and literature review. AB - CONTEXT: Intussusception in adults is rare, accounting for 0.1% of adult hospital admissions. In contrast to this, it is the leading cause of obstruction in children. In up to 90% of adults a cause can usually be found, but in children this is rarely the case. CASE REPORT: We report the case of a 27-year-old lady with a subacute bowel obstruction caused by a jejunal heterotopic pancreas and describe its successful surgical management. CONCLUSION: The causes of intussusception in adults are discussed in the literature review. PMID- 15536286 TI - An unusual case of gastric heterotopic pancreas. AB - CONTEXT: Submucosal lesions of the gastrointestinal tract represent a diagnostic challenge for the physician. Endoscopic ultrasonography may provide useful information before deciding on therapeutic strategy. CASE REPORT: We report on a case of a young female presenting with a large gastric submucosal mass, 32 mm in size. Endoscopic ultrasonography identified a non-homogeneous lesion, with three cystic spaces suggesting a degenerated gastrointestinal stromal tumor. An exploratory laparoscopy was performed. Surprisingly, the final diagnosis was gastric heterotopic pancreas. CONCLUSION: Heterotopic pancreas should always be kept in mind when facing extramucosal gastric masses, especially in young people. A perioperative biopsy is recommended to prevent unnecessary extensive surgery. PMID- 15536285 TI - Discrepancy between pancreatographic and histopathological findings in the ventral pancreas of pancreas divisum. AB - CONTEXT: Pancreas divisum is a congenital anomaly in which the ventral and dorsal pancreatic ducts do not communicate. CASE REPORT: Autopsy case of pancreas divisum with a history of heavy consumption of alcohol was presented. Pancreatography via the major duodenal papilla showed a short ventral pancreatic duct of 1 cm in length, and hypoplasia of the ventral pancreas was suspected. As large cysts were confined to the dorsal pancreas, isolated dorsal pancreatitis was also suspected. At autopsy, extensive fibrosis was detected in both the ventral and dorsal pancreas. Immunohistochemical examination revealed that PP rich islets were scattered in the fibrotic area between the ventral pancreatic parenchyma and the neck of the pancreas, suggesting that this fibrotic area originated from the ventral pancreas. These facts suggest that the short ventral pancreatic duct was not induced by hypoplasia of the ventral pancreas but was shortened secondarily by the alcohol-induced fibrosis. CONCLUSIONS: In pancreas divisum, a short ventral pancreatic duct resulting from secondary factors may be confused with that originating from hypoplasia of the ventral pancreas. PMID- 15536287 TI - Pitfalls of MRCP in the diagnosis of pancreaticobiliary maljunction. AB - CONTEXT: Magnetic resonance cholangiopancreatography (MRCP) is useful for examining the pancreatic duct system in patients with acute pancreatitis instead of using endoscopic retrograde cholangiopancreatography (ERCP), as ERCP-induced pancreatitis represents a serious problem. However, we present here a case of idiopathic acute pancreatitis in which MRCP suggested pancreaticobiliary maljunction, but ERCP indicated normal pancreaticobiliary union. CASE REPORT: A 22-year-old male was urgently admitted complaining of upper abdominal and back pain. He had no history of alcohol or drug intake. Serum amylase levels were elevated to 880 U/mL (reference value: less than 158 U/mL). Abdominal ultrasound demonstrated only a slight swelling of the pancreas, but no abnormal findings for the bile duct or gallbladder. Symptoms and hyperamylasemia improved with supportive therapy. Coronal heavily T2-weighted single-shot rapid acquisition with relaxation enhancement MRCP indicated a markedly long common channel, and pancreaticobiliary maljunction without biliary dilatation was diagnosed. Under the diagnosis of idiopathic acute pancreatitis associated with pancreaticobiliary maljunction without biliary dilatation, prophylactic laparoscopic cholecystectomy was planned. However, ERCP demonstrated a narrow main pancreatic duct and a normal common bile duct without the formation of a common channel. In a supine position, after withdrawal of the scope, the narrow main pancreatic duct at the head of the pancreas overlapped the lower common bile duct, giving the appearance of a long common channel as indicated by MRCP. CONCLUSIONS: In MRCP of cases with a narrow main pancreatic duct, there is a possibility for false-positive indications of pancreaticobiliary maljunction. MRCP with secretin stimulation or ERCP should be performed in such cases. PMID- 15536288 TI - Peripancreatic Castleman disease. AB - CONTEXT: Castleman disease or giant lymph node hyperplasia is a rare disorder of the lymphoid tissue, usually occurring in the mediastinum. We report a case of localized peripancreatic plasma cell type Castleman disease, which is an unusual site. CASE REPORT: A 45-year-old woman with a history of mild epigastric pain radiating to the back for the previous year was admitted and her physical examination was normal. A raised erythrocyte sedimentation rate, C-reactive protein and hypergammaglobulinemia were found. Abdominal ultrasonography and magnetic resonance imaging demonstrated a homogeneously hypoechoic solid mass having a smooth-surface, 6 cm in diameter, located between the head of the pancreas, the lower margin of the antrum and the left lobe of the liver. The mass was totally excised surgically with repair of the pancreas capsule. Histopathological examination of the tumor showed plasma-cell type Castleman disease. CONCLUSION: Clinicians should be aware that Castleman disease may involve peripancreatic tissue which leads to difficulties in arriving at a differential diagnosis. Surgical excision is both a diagnostic and a curative method for management of the disease. PMID- 15536289 TI - Signet-ring-cell carcinoma of the Vater's ampulla. AB - CONTEXT: Most tumors affecting Vater's Ampulla are adenocarcinomas and other histological variants are less frequent. A review of the literature revealed only seven previously reported cases of signet ring cell carcinoma of the ampulla of Vater. The presence of this kind of tumor has no clear histological explanation. Two possible theories have been proposed: the presence of gastric heterotopia in the ampulla of Vater or the existence of a perivaterian duodenal heterotopia of ulcerous etiology as the origin of a signet ring cell tumor which secondarily invades the ampulla of Vater. CASE REPORT: We performed a pancreatoduodenectomy in a 67-year-old woman with a T2N0M0 ampulla tumor. A histologic study revealed a signet ring cell neoplasm. CONCLUSION: Etiology and survival of signet ring cell carcinoma of Vater's ampulla is not well-defined in the literature due to the extreme rarity of this disease. Duodenopancreatectomy with pylorus preservation is the treatment of choice. PMID- 15536290 TI - Pseudocyst formation in gastric ectopic pancreas. AB - CONTEXT: It is rare for ectopic pancreatic tissue to cause symptoms or require treatment, however diseases of normal pancreas may also occur in ectopic pancreas tissue. CASE REPORT: This report describes the clinical, endoscopic, radiologic and histologic features of a pseudocyst occurring in gastric ectopic pancreas in a 19-year-old man. The difficulty and implications of making an accurate pre operative diagnosis are highlighted. CONCLUSION: Ectopic pancreatic tissue, although rare, should be considered in the differential diagnosis of a submucosal gastric mass. PMID- 15536291 TI - Recurrent acute pancreatitis possibly induced by atorvastatin and rosuvastatin. Is statin induced pancreatitis a class effect? AB - CONTEXT: Few data exist about the incidence of drug-induced pancreatitis in the general population. Drugs are related to the etiology of pancreatitis in about 1.4-2% of cases. While statins are generally well tolerated they have been known to be associated with pancreatitis. Acute pancreatitis has been reported in a few cases treated with atorvastatin, fluvastatin, lovastatin, simvastatin and pravastatin. CASE REPORT: We report the case of a 77-year-old patient who developed acute pancreatitis after treatment with rosuvastatin, which resolved on withdrawal of the medication. She had a similar episode of pancreatitis a year ago precipitated by atorvastatin, which resolved on withdrawal. Extensive workup on both occasions failed to reveal any other etiology for the pancreatitis. CONCLUSION: To our knowledge this is the first report of rosuvastatin-induced pancreatitis. The occurrence of pancreatitis with two different statins in our patient argues that statins induced pancreatitis may be a class-effect of statins. With statin prescriptions on the rise clinicians need to be aware of this complication of statin treatment and remember that the newest statin, rosuvastatin is not dissimilar to the other statins in causing pancreatitis. PMID- 15536292 TI - Pancreatic pseudopapillary tumor in a male child. AB - CONTEXT: Solid-pseudopapillary tumors are exceedingly rare in males. They are almost exclusively encountered in young females (mean age 26 years) and have a female predominance. It is most commonly detected incidentally, but may occasionally present with sudden pain or symptoms related to compression of adjacent organs. CASE REPORT: We report the case of a 12-year-old boy having a solid-pseudopapillary tumor of the pancreas presenting with a tender upper abdominal mass following a slight trauma. Radiological investigation showed the lesion to be a cystic mass arising from the body and the tail of the pancreas. The child underwent emergency distal pancreatectomy and has remained free of recurrence for 3 years. CONCLUSION: In the pediatric age group, solid pseudopapillary tumors may present acutely with a tender abdominal mass following a slight trauma. Awareness of this fact will allow appropriate and prompt management to be undertaken. PMID- 15536293 TI - Anaplastic pancreatic carcinoma. A case report and review of literature . AB - CONTEXT: Anaplastic pancreatic carcinoma is an aggressive neoplasm with survival measurable in weeks. It presents as a large cystic mass with loco-regional and distant spread. Three histological types have been described: pleomorphic, spindle cell and sarcomatoid. CASE REPORT: We describe the case of a 74-year-old woman with pleomorphic anaplastic carcinoma of the pancreas diagnosed after laparoscopic biopsy. The patient had a rapid downhill course with progression of the disease and demise within 4 weeks after diagnostic laparoscopy. CONCLUSION: Due to the rapid spread of the disease, no effective cure exists for these tumors. A brief review of the histological and radiological findings and the possible mechanisms of the pathogenesis of anaplastic tumors is included in the discussion. PMID- 15536294 TI - Metronidazole-induced pancreatitis. A case report and review of literature. AB - CONTEXT: Pancreatitis is a very rare adverse effect of metronidazole with only six cases of metronidazole-induced pancreatitis reported in the English literature so far. CASE REPORT: We report a case of recurrent acute pancreatitis in a 46-year-old female associated with oral metronidazole therapy and review the literature with regards to metronidazole-induced pancreatitis. We are also highlighting the fact that the time lag between metronidazole exposure and development of pancreatitis is very variable. CONCLUSION: High degree of suspicion is warranted on the part of physicians to diagnose metronidazole induced pancreatitis in patients presenting with gastrointestinal symptoms after metronidazole exposure. If metronidazole is suspected as the causative agent then it should be discontinued and rechallenge should be avoided. PMID- 15536295 TI - Pancreatic schwannoma. A case report and review of the literature. AB - CONTEXT: Pancreatic schwannomas are rare neoplasms. These tumors vary in size and two thirds are partially cystic which grossly mimic pancreatic cystic lesions. Computed tomography and magnetic resonance imaging are the primary initial imaging modalities. Definitive diagnosis is typically made at the time of laparotomy following biopsy. Surgical resection is the mainstay of treatment. CASE REPORT: A 69-year-old woman presented with abdominal pain in the epigastric and left upper quadrant. The patient had no systemic symptom and laboratory results including tumor markers were negative. A CT scan of the abdomen showed a 5 cm mass arising from the head of the pancreas. Needle biopsy revealed a mass consistent with schwannoma. At laparotomy, a large pancreatic head mass was found to encase the superior mesenteric artery, and portal vein confluence. Frozen biopsy showed schwannoma. Curative resection was deferred due to extensive vascular involvement and favorable tumor biology. A gastrojejunostomy was performed and radiation therapy was instituted post-operatively. CONCLUSIONS: Only 24 cases of pancreatic schwannoma had been previously reported. Definitive diagnosis is obtained with routine histology. Most tumors are benign and surgical resection is curative. The role of radiation therapy in the management of unresectable tumors is still unclear. PMID- 15536297 TI - IgG4 as a serological marker of autoimmune pancreatitis: the latest news. PMID- 15536296 TI - Autoimmune pancreatitis. A case mimicking carcinoma. PMID- 15536298 TI - Case reports and case studies: an endangered species. PMID- 15536299 TI - The unknown source of John Hughlings Jackson's early interest in aphasia and epilepsy. AB - The National Hospital for Paralysis and Epilepsy in London (founded 1859) was the scene of great discoveries in the new specialty of neurology, carried out in great part by John Hughlings Jackson (1834-1911). The clinicians Jonathan Hutchinson and Charles Edward Brown-Sequard are typically identified as Jackson's mentors. This paper discusses the previously neglected role of Jabez Spence Ramskill (1824-1897), founding physician of the National Hospital. Ramskill appears to have been significant in providing the opportunity and context that led Jackson to develop his theories concerning higher cerebral function disorders. As assistant physician to Ramskill at the National Hospital, Jackson was provided with a vast caseload of epileptic, hemiplegic, and aphasic patients. Ramskill and Jackson both published papers on aphasia in the London Hospital Reports in 1864. Consideration of the similarities and differences between these 2 papers highlight significant issues in the clinical and theoretical development of understanding language organization in the brain. The early writings of Jackson and case notes of Ramskill document a close link between the 2 and indicate the debt that Jackson had to Ramskill for providing him with the opportunities to develop his original ideas on epilepsy and aphasia. PMID- 15536300 TI - Alterations of the sense of "humanness" in right hemisphere predominant frontotemporal dementia patients. AB - OBJECTIVE: To evaluate the sense of "humanness" in frontotemporal dementia (FTD) patients with right hemispheric involvement. BACKGROUND: Early in the course, FTD is often asymmetric, and those with predominant right frontotemporal disease have disproportionate disturbances in social behavior and empathy. A disruption in a sense of humanness may underlie these behavioral disturbances. METHOD: Sixteen patients with asymmetric FTD on functional neuroimaging underwent recognition tests of facial masking, human-animal morphing, and facial distortion. Additional tests evaluated facial discrimination and the recognition of famous faces, facial emotions, and animate-inanimate differences. RESULTS: On the distorted and morphed face tasks, 8 FTD patients with predominant right hemisphere involvement were significantly more likely to call morphed and distorted faces "human" as compared with both 8 FTD patients with predominant left hemisphere involvement and normal controls. The FTD groups did not differ on thresholds for recognizing masked faces or on other face recognition measures. CONCLUSIONS: In FTD, right hemispheric involvement may alter the threshold for judging someone as human independent of the recognition of faces or facial affect. These results suggest that a specific sense of humanness facilitates a person recognition network in the right frontotemporal region of the brain. PMID- 15536301 TI - Prevalence and clinical correlates of disinhibition in dementia. AB - OBJECTIVE AND METHODS: The phenomenology, main clinical correlates, and long-term evolution of disinhibition in dementia are not well known. To examine this issue, we studied a consecutive series of 272 patients with probable Alzheimer disease using a comprehensive psychiatric and neuropsychological evaluation that included the Disinhibition Scale. A subset of patients was reexamined with the same instruments between 1 and 4 years after the initial evaluation. RESULTS: A factor analysis of the Disinhibition Scale demonstrated 4 factors: (1) abnormal motor behavior, (2) hypomania, (3) loss of insight and egocentrism, and (4) poor self care. Disinhibition was significantly associated with major and dysthymic depression, more severe negative symptoms, and loss of awareness. Most patients with disinhibition at the initial evaluation still showed disinhibition at follow up, whereas 23% of patients without disinhibition at the initial evaluation developed disinhibition at follow-up. CONCLUSIONS: Disinhibition is a frequent and long-lasting problem in dementia. Our study demonstrates that the construct of disinhibition consists of 4 independent subsyndromes, each of which may have specific underlying mechanisms. PMID- 15536302 TI - Self-reported memory symptoms with coronary artery disease: a prospective study of CABG patients and nonsurgical controls. AB - OBJECTIVE: Subjective memory complaints are common after coronary artery bypass grafting (CABG), but previous studies have concluded that such symptoms are more closely associated with depressed mood than objective cognitive dysfunction. We compared the incidence of self-reported memory symptoms at 3 and 12 months after CABG with that of a control group of patients with comparable risk factors for coronary artery disease but without surgery. METHODS: Patients undergoing CABG (n = 140) and a demographically similar nonsurgical control group with coronary artery disease (n = 92) were followed prospectively at 3 and 12 months. At each follow-up time, participants were asked about changes since the previous evaluation in areas of memory, calculations, reading, and personality. A Functional Status Questionnaire (FSQ) and self-report measure of symptoms of depression (CES-D) were also completed. RESULTS: The frequency of self-reported changes in memory, personality, and reading at 3 months was significantly higher among CABG patients than among nonsurgical controls. By contrast, there were no differences in the frequency of self-reported symptoms relating to calculations or overall rating of functional status. After adjustment for a measure of depression (CES-D rating score), the risk for self-reported memory changes remained nearly 5 times higher among the CABG patients than among control subjects. The relative risk of developing new self-reported memory symptoms between 3 and 12 months was 2.5 times higher among CABG patients than among nonsurgical controls (CI 1.24-5.02), and the overall prevalence of memory symptoms at 12 months was also higher among CABG patients (39%) than controls (14%). CONCLUSIONS: The frequency of self-reported memory symptoms 3 and 12 months after baseline is significantly higher among CABG patients than control patients with comparable risk factors for coronary and cerebrovascular disease. These differences could not be accounted for by symptoms of depression. The self reported cognitive symptoms appear to be relatively specific for memory and may reflect aspects of memory functioning that are not captured by traditional measures of new verbal learning and memory. The etiology of these self-reported memory symptoms remains unclear, but our findings, as well as those of others, may implicate factors other than cardiopulmonary bypass itself. PMID- 15536303 TI - Persistent severe depressive episode with mood-congruent psychotic features associated with left temporal ischemia. AB - OBJECTIVE: The authors describe the case of a patient suffering from a persistent severe depressive episode with mood-congruent psychotic features following left temporal ischemia. BACKGROUND: Depression is a frequent disorder that may stem from psychosocial factors or occur after brain damage. Cases of poststroke depression with associated psychotic symptoms are rarely described in the literature, and, to our knowledge, no case of depression with psychotic symptoms in the presence of a focal lesion has yet been reported. METHOD: History included psychiatric, neurologic, and neuropsychological as well as EEG data and CT scan and magnetic resonance images. RESULTS AND CONCLUSIONS: Neurologic and psychiatric dimensions may be associated and difficult to disentangle, even when the patient presents the classic symptoms of a depressive disorder. This case underlines the importance of investigating possible neurologic signs or symptoms, even when the psychiatric presentation is prototypical of a determined mental disorder. This may be crucial for the patient's evolution. PMID- 15536304 TI - A high resolution assessment of the repeatability of relative location and intensity of transcranial magnetic stimulation-induced and volitionally induced blood oxygen level-dependent response in the motor cortex. AB - OBJECTIVE: Using functional magnetic resonance imaging, we assessed variation in location and intensity of blood oxygen level-dependent contrast associated with movements induced by transcranial magnetic stimulation or volition. BACKGROUND: Anatomic location and within-subject repeatability of blood oxygen level dependent responses induced by transcranial magnetic stimulation comprise critical information to the use of interleaved transcranial magnetic stimulation/functional magnetic resonance imaging as a neuroscience tool. METHODS: Eleven healthy adults were scanned 3 times each at 1.5 T. Interleaved with functional magnetic resonance imaging, 1-Hz transcranial magnetic stimulation was applied over motor cortex. VOL was alternated with transcranial magnetic stimulation over the scans. RESULTS: Intra-subject standard deviations in blood oxygen level-dependent locations ranged between 3 and 6 millimeters, allowing localization to subregions of the motor strip. Coil placement relative to blood oxygen level-dependent location varied more than blood oxygen level dependent location (sdx = 9.5mm, sdy = 8.7 mm, sdz = 9.0mm) with consistent anterior displacement (dy = 21.8 mm, P = <0.025). Analysis of variance did not detect significant differences between transcranial magnetic stimulation and VOL blood oxygen level-dependent locations or intensities, in contrast to significant intensity differences detected in auditory blood oxygen level dependence. CONCLUSION: The high repeatability of location of transcranial magnetic stimulation-induced blood oxygen level-dependent activation suggests that transcranial magnetic stimulation/functional magnetic resonance imaging stimulation can be used as a precise tool in investigation of cortical mechanisms. The similarity between VOL and transcranial magnetic stimulation suggests that transcranial magnetic stimulation may act through natural brain movement circuits. PMID- 15536305 TI - Pseudoneglect in solid-line versus character-line bisection tasks: a test for attention dominance theory. AB - OBJECTIVE: Normal subjects tend to bisect lines slightly to the left of the true midpoint, a phenomenon termed pseudoneglect. To test whether pseudoneglect relates to the right hemisphere's dominance for spatial attention or to the hemispheric difference in processing global-local stimulus properties, we administered conventional solid-line (SBT) and novel character-line (CBT) bisection tasks to normal subjects of different ages. METHODS: Normal subjects, consisting of 40 young and 40 older individuals, received 3 experimental tasks, a standard SBT and 2 types of CBT. Each subject completed 10 consecutive trials of each task presented in counterbalanced order between subjects. RESULTS: Across age groups, deviations on CBT were further to the right than those of SBT, and the leftward bias (pseudoneglect) was significant only in SBT. CONCLUSION: These results indicate that the bisection errors in normal subjects depend on the characteristics demanded by the specific task. Thus, our findings argue against the attention dominance theory and support a "task specificity" theory for pseudoneglect. PMID- 15536306 TI - Cognitive recovery in idiopathic normal pressure hydrocephalus after shunt. AB - OBJECTIVE: Evaluate long-term changes in cognition after ventriculoperitoneal shunt insertion in patients with idiopathic normal pressure hydrocephalus. BACKGROUND: Although studies have indicated that treatment of normal pressure hydrocephalus with ventriculoperitoneal shunt improves gait, effects of treatment on cognition have been inconsistent. Several studies have reported no significant improvement on neuropsychological tests in patients with idiopathic normal pressure hydrocephalus. Selection of candidates for shunt surgery using a trial of controlled spinal fluid drainage via lumbar catheter may result in more consistent improvement in cognition. DESIGN/METHODS: This was a prospective, longitudinal study of performance on neuropsychological tests before and 6 to 12 months after insertion of a ventriculoperitoneal shunt in 10 patients with idiopathic normal pressure hydrocephalus who showed improvement in at least 1 clinical symptom with temporary lumbar drainage. Nonparametric tests were used to evaluate change. RESULTS: Significant group improvements at follow-up were demonstrated on tests of verbal memory as well as in 1 test of psychomotor speed. Eight of 10 patients showed improvement by more than 1 standard deviation on at least 1 memory test. Six of 10 patients improved significantly on more than 50% of the tests administered. CONCLUSIONS: Patients with idiopathic normal pressure hydrocephalus who show clinical improvement (in any symptom) after lumbar drainage are likely to show significant long-term improvement in memory after ventriculoperitoneal shunt. PMID- 15536307 TI - Free transverse rectus abdominis musculocutaneous flap breast reconstruction in patients with prior abdominal suction-assisted lipectomy. PMID- 15536308 TI - Current concepts in aesthetic upper blepharoplasty. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to: 1. Discuss nomenclature and anatomy associated with upper blepharoplasty. 2. Perform preoperative assessment, decision-making, and counseling of patients. 3. Describe current surgical planning, eyelid marking, and various techniques used in upper blepharoplasty, including lasers. 4. Recognize and treat postoperative complications from blepharoplasty. Traditional blepharoplasty has often involved the excision of both lax skin and muscle and excessive removal of fat, leaving patients long term with a hollow orbit and a harsh, operated appearance that accelerates the aging process. Current methods of periorbital rejuvenation are more conservative, are based on concise preoperative evaluation, and involve the limited resection of the coveted soft tissue from the eye to restore a youthful appearance. The authors describe anatomy, preoperative assessment, decision making and counseling of patients, surgical planning, eyelid marking, and various techniques, including lasers, along with postoperative complications associated with current concepts in aesthetic upper blepharoplasty. PMID- 15536326 TI - Comparison of 34betaE12 and P63 in 100 consecutive prostate carcinoma diagnosed by needle biopsies. AB - P63, a homologue of p53, was recently identified as a useful basal cell-specific marker. We compared the sensitivity and specificity of p63 with the widely used high-molecular-weight keratin 34betaE12 for the diagnosis of prostate carcinoma in needle biopsies. We selected 100 consecutive prostate carcinoma diagnosed by needle biopsies with an adequate number of cancerous glands on the slide. We chose 1 representative hematoxylin and eosin-stained slide from each case and gave it a Gleason score. The same paraffin block was retrieved for 34betaE12 and p63 stains. We compared staining patterns of 34betaE12 and p63 on both malignant glands and benign glands and recorded basal cell density (percentage of basal cells with positive staining in the benign glands). The cases were divided into 3 groups according to the Gleason score: 5 to 6 (31 cases), 7 (46 cases), and 8 to 10 (23 cases). In 20 cases, focal and patchy staining in a basal cell distribution in malignant glands (range, 1%-20%; mean, 6.6%) was demonstrated (19 by both stains and 1 by 34betaE12 only). In 1 case with a Gleason score of 9, the cancer cells, not the basal cells, were stained focally by p63 but not by 34betaE12. Higher-grade tumors demonstrated higher numbers of malignant glands with basal cell staining (1.65% for Gleason 7, 1.26% for Gleason 8-10, compared with 0.42% for Gleason 5-6). The overall specificity of the absence of basal cell staining in the malignant glands for 34betaE12 and p63 was 98.63% and 98.60%, respectively. In 17 cases, both stains revealed total absence of basal cell staining in some benign glands (range, 1%-10%; mean, 3.5%). The overall sensitivity in identifying basal cells in benign glands was 99.48% and 99.44% for 34beta12 and p63, respectively. Basal cell density was higher for 34betaE12 in comparison with p63 (92% vs. 87%). For diagnosing prostate carcinoma in the needle biopsies, p63 is as specific and sensitive Hospital as 34betaE12 and therefore can be used as a complementary basal cell-specific stain for 34betaE12 in difficult cases. PMID- 15536327 TI - MIB1 labeling index as an indicator of chemoresponse in carcinoma of the breast. AB - In this study, we determined the extent of variation in proliferative markers and hormone receptor status in breast carcinoma between core biopsies and subsequent resections, and determined the impact of clinical and histologic parameters on such variation. We performed a paired comparison of biomarkers in 87 core biopsies and subsequent resections of breast carcinomas in patients with and without preoperative chemotherapy. The markers included estrogen receptor, progesterone receptor, Her2/neu, DNA ploidy (diploidy versus nondiploidy), DNA index (difference of > or = 0.5), and MIB1 labeling index (<15% vs. >15%). The tumor biomarkers were evaluated with standard IHC and scored by automated cellular imaging systems. The number of patients showing prominent changes were as follows: ploidy, 12; DI, 15; ER, 6; PR, 15; MIB1, 17; and Her2/neu, 15. Seventeen of 87 patients sustained a significant change in MIB1 index (above or below 15%). The patients who received chemotherapy had a larger proportion with a change in MIB1 status (P < 0.05). DI showed a similar trend, although it was not statistically significant. Of the patients with MIB1 values higher in biopsy specimens, a majority showed values to decrease below 15%. MIB1 index reduction by greater than 25% was seen in 8 of 16 (50%) cases with chemotherapy, compared with only 3 of 32 (9%) without chemotherapy. MIB1 and DNA index values postchemotherapy can be a useful measure of chemoresponse. Our study underscores the need to perform prognostic markers on both biopsy and subsequent resections, especially in the setting of preoperative chemotherapy. PMID- 15536328 TI - Analysis of survivin expression in a spectrum of benign to malignant lesions of the breast. AB - Survivin, a novel inhibitor of apoptosis, is expressed in a variety of human cancers, with reports of prognostic significance in some neoplasms. The authors' aim was to evaluate survivin expression in a spectrum of breast lesions to determine differential expression in malignant versus benign lesions and its potential role as a diagnostic or prognostic marker. The authors found that survivin is expressed in breast tissue in the full spectrum of normal to invasive carcinoma. It is predominantly nuclear with a faint cytoplasmic blush. Survivin expression was independent of patient age and tumor size. Benign breast tissue showed survivin expression in a lower percentage of cells (45%) than malignant lesions. The median values for the percentage of cells that stained for survivin were statistically different among the categories of invasive carcinoma, DCIS, LCIS, and benign breast tissue (P < or = 0.001). The highest percentage of positive-staining cells was seen in high-grade DCIS (95%). The authors found a trend toward a higher percentage of cells staining for survivin in breast carcinoma cases that were ER negative, PR negative, or Her2/neu positive, although this was not statistically significant. Survivin expression was preserved in biopsies from recurrent tumors without loss of nuclear survivin expression. In conclusion, survivin is overexpressed in malignant breast lesions relative to benign lesions or normal breast tissue and in high-grade DCIS relative to nonhigh-grade DCIS. Therefore, survivin may have a role, albeit a limited one, as a prognostic marker in breast lesions. PMID- 15536329 TI - Morphologically similar epithelial and stromal cells in primary bilateral breast tumors display different genetic profiles: implications for treatment. AB - The morphologic features of primary bilateral breast carcinoma have been well elucidated, but it is not known whether tumors at two sides share a common genetic profile and undergo the same clinical course. To address this issue, morphologically comparable epithelial and stromal cells in 18 paired primary bilateral breast tumors were microdissected and subjected to comparisons for the frequency and pattern of loss of heterozygosity (LOH) and microsatellite instability (MI), as well as the profiles of comparative genomic hybridization. Of 18 paired bilateral epithelial samples assessed with 10 DNA markers at five chromosomes, 78 altered loci were found; of these, 23 (29.5%) displayed concurrent and 55 (70.5%) showed independent LOH, MI, or both. Of 18 paired bilateral stromal samples assessed with the same markers, 70 altered loci were seen; of these, 9 (12.9%) displayed concurrent and 61 (87.1%) showed independent LOH, MI, or both. Collectively, all the markers and 30 (83.3%) of 36 paired bilateral epithelial and stromal cells displayed significantly more (P < 0.01) independent than concurrent LOH, MI, or both. In contrast, the epithelial cells of a pulmonary small cell carcinoma metastasized to both breasts displayed concurrent LOH at each of the four altered loci. Of seven selected cases for comparative genomic hybridization, six (86%) displayed chromosomal changes, but none showed an identical pattern and frequency of changes in both breasts. The significantly higher rate of independent genetic alterations in morphologically comparable cells of paired bilateral primary breast tumors supports the notion that the development and clinical course of tumors in two sides differ substantially; consequently, different interventions might be needed for the optimal management of bilateral breast tumors. PMID- 15536330 TI - CYP2E1 polymorphism, cigarette smoking, p53 expression, and survival in non-small cell lung cancer: a long term follow-up study. AB - The expression of selected gene products involved in cell differentiation and cell growth and genetic polymorphism of detoxifying genes was examined in 105 surgically resected nonsmall cell lung cancer (NSCLC) patients, and the relationship of these factors was correlated with cigarette smoking and patient survival. Genotyping of peripheral blood lymphocytes from 87 patients was performed for CYP2E1, GSTM1, GSTT1, mEH, and MPO detoxifying genes using polymerase chain reaction. Formalin-fixed, paraffin-embedded tissue was immunostained with antibodies to p53, p27, phospho-AKT, and bcl-2 using the avidin-biotin-peroxidase method and tissue microarray technique. Tumors were assigned a positive or negative score based on more than 10% of tumor cells staining positive with the antibody. The subtypes of NSCLC included 48 adenocarcinomas, 47 squamous cell carcinomas, and 10 large cell undifferentiated carcinomas. A total of 54 tumors were pathologic stage I, 23 were stage II, and 26 were stage III. All subjects smoked (range, 10-175 pack-years; mean, 60 pack years). The mean overall survival was 112 weeks (median, 129 weeks). Patients with p53-positive tumors had significantly fewer pack-years of smoking (52 pack years vs 72 pack-years; P = 0.021), smoked fewer years (34 years vs 40 years; P = 0.018), and had significantly better survival compared with those with p53 negative tumors (P = 0.045). When smoking history was further analyzed, the authors found that p53 expression was associated with the number of years smoked and not the number of packs smoked per day. Patients with squamous cell carcinoma had smoked longer compared with those with adenocarcinoma (P = 0.011). Significant association was seen between the CYP2E1 wild-type allele and better survival (P = 0.016). Patients with stage I tumors had better survival compared with stages II and III (P = 0.032). No association was found between survival and tumor type; tumor differentiation; expression of phospho-AKT, p27, and bcl-2; and polymorphic metabolizing genes other than CYP2E1. The significant association of long duration of smoking (>40 years) with loss of p53 expression and poor survival suggests inactivation of the protective p53 pathway in those who had a history of more than 40 years of smoking. PMID- 15536331 TI - Prognostic significance of dysadherin expression in tongue cancer: immunohistochemical analysis of 91 cases. AB - The E-cadherin-mediated cell adhesion system is frequently inactivated by multiple mechanisms and is involved in tumor progression in many types of cancer. Recently we have reported a novel cell membrane glycoprotein, dysadherin, which has an anti-cell-cell adhesion function and downregulates E-cadherin. Expressions of dysadherin and E-cadherin were investigated immunohistochemically in 91 patients with squamous cell carcinoma of the tongue to determine the correlation between the 2 molecules and their associations with the clinicopathologic features of the tumors and with patient survival. Dysadherin was expressed at the cell membranes of many cancer cells. Twenty-five percent of the tumors showed dysadherin immunopositivity in more than 50% of the cancer cells. Sixty-nine percent of the tumors showed reduced E-cadherin immunopositivity. There was an inverse correlation between dysadherin expression and E-cadherin expression (P = 0.0001). Increased dysadherin expression was significantly correlated with an infiltrative type of growth pattern (P = 0.001), high tumor-node-metastasis (TNM) stage (P = 0.024), and poor patient survival (P = 0.003). After adjusting for growth pattern, TNM stage, and other clinicopathologic features, increased dysadherin expression and reduced E-cadherin expression were both significant predictors of poor survival (P = 0.0006). Increased dysadherin expression is a significant indicator of poor prognosis in patients with squamous cell carcinoma of the tongue. PMID- 15536332 TI - Pathologic study and clinical significance of Hurthle cell papillary thyroid carcinoma. AB - Hurthle cell papillary thyroid carcinoma is a variant of papillary thyroid carcinoma (PTC). Its pathologic and clinical significance has not been well documented. The authors studied the relative incidence of Hurthle cell PTC and the relationship of Hurthle cell PTC to other variants of thyroid carcinoma. Three hundred eighty consecutive cases of thyroid carcinoma were reviewed to identify cases with focal or extensive areas of Hurthle cell PTC, classic PTC, Hurthle cell carcinoma (ie, non-Hurthle cell PTC), and follicular carcinoma. In addition, the status of lymphoid infiltrate in the tumor, stromal invasion with desmoplastic reaction, vascular invasion, and distant and lymph node metastasis were noted by microscopic examination, review of clinical charts, or both. A total of 24 (HCs) and 42 PTCs with Hurthle cells were identified. The latter category was divided into pure Hurthle cell PTC or extensive Hurthle cell (HPTC) (28 cases) and PTC or Hurthle cell carcinoma with focal areas of Hurthle cell PTC (14 cases). The Hurthle cell PTC/Hurthle cell carcinoma ratio was lower than that of PTC/follicular carcinoma (39:289) (P = 0.001). Follicular or solid structures were present in all HPTCs. HPTCs were associated with frequent stromal intrathyroid and extrathyroid invasion, but they tended to have a lower rate of lymph node metastasis (8/28) compared with classic PTC with stromal invasion (108:200) (P = 0.12) and a lower rate of distant metastasis (2:28) compared with Hurthle cell carcinoma (15:24) (P = 0.02) or follicular carcinoma (13:39) (P = 0.04). Warthin-like Hurthle cell PTC (10 cases) was associated with extrathyroid invasion in five cases. In Hurthle cell PTC associated with tall cell variant (10 cases), areas of gradual transition between Hurthle cell PTC and tall cell variant were identified. The latter variant showed the highest rate of extrathyroid stromal and vascular invasion with distant metastasis and patient death compared with all Hurthle cell PTCs and classic PTCs. In conclusion, Hurthle cell PTC is frequently associated with tall cell variant. It has a higher potential for extrathyroid invasion than classic PTC and has vascular invasion and distant metastasis characteristics intermediate between those of classic PTC and Hurthle cell carcinoma with or follicular carcinoma. Hurthle cell PTC tends to show a greater likelihood of extrathyroid invasion when associated with Warthin-like features and tall cell variant PTC, and higher vascular invasion and distant metastasis when associated with tall cell variant. PMID- 15536333 TI - Estrogen and progesterone receptor expression in uterine and extrauterine leiomyosarcomas: an immunohistochemical study. AB - The authors have noted anecdotal cases of extrauterine leiomyosarcomas (LMS) with estrogen receptor (ER) and progester-one receptor (PR) immunoreactivity. However, there are few studies that have compared ER and PR immunoexpression in LMS of uterine and extrauterine origin. The authors obtained a representative formalin fixed, paraffin-embedded tissue block from cases of uterine LMS (n = 15) and extrauterine LMS (n = 16) from the archives of the Cleveland Clinic Foundation and performed immunohistochemical staining for ER and PR. Staining was evaluated by 2 observers in a semiquantitative manner using the following scale: 0, no nuclear staining; 1+, 1 to 25% of nuclei stained; 2+, 26 to 50% of nuclei stained; 3+, 51 to 75% of nuclei stained; 4+, 76 to 100% of nuclei stained. The majority of uterine LMS stained for ER (13 of 15, 87%), PR (12 of 15, 80%), or both ER and PR (12 of 15, 80%), with most cases showing 3+ or 4+ positive staining. For the extrauterine LMS cases, staining for ER was seen in 4 of 16 cases (25%), staining for PR was observed in 2 of 16 cases (13%), and staining for both ER and PR was seen in 2 of 16 cases (13%). One extrauterine LMS showed 4+ coexpression of ER and PR, but the remaining extrauterine cases showed only 1+ ER and/or PR immunoreactivity. These data suggest that most uterine LMS coexpress ER and PR, and most extrauterine LMS do not stain for these antigens. However, a subset of extrauterine LMS are ER and/or PR immunoreactive. This raises the possibility that hormonal manipulation may be beneficial in the treatment of these therapeutically recalcitrant tumors. PMID- 15536334 TI - Focal adhesion kinase overexpression in endometrial neoplasia. AB - Focal adhesion kinase (FAK) is a protein tyrosine kinase that is a critical mediator of signaling events between cells and their extracellular matrix. Elevations in FAK mRNA and protein overexpression have been linked to tumor cell capacity for invasion and metastasis. FAK expression has been shown to be elevated in a variety of solid tumors. The purpose of this study was to evaluate for FAK upregulation in endometrial neoplasia. Tissue microarray blocks were made from formalin-fixed, paraffin-embedded archival tissue, including 115 carcinoma (100 endometrioid, 10 serous, and 5 clear cell), 28 hyperplasia, and 38 normal specimens using 1-mm punches. The tissue was immunostained with monoclonal antibody for FAK and p53. Immunoreactivity was scored by intensity (0-4+ scale) and percent positive staining. FAK overexpression was categorized as 4+ cytoplasmic intensity in more than 90% of neoplastic cells. Positive p53 was categorized at least 2+ nuclear intensity in more than 10% of neoplastic cells. Higher rates of FAK upregulation were identified in endometrial hyperplasia (P = 0.025) and carcinoma (P < 0.001) versus normal endometrium. FAK overexpression in carcinoma correlated with higher FIGO grade (P = 0.025) and p53 overexpression (P < 0.001). FAK was consistently overexpressed in high-grade tumors regardless of subtype, including 8 of 10 serous tumors, 4 of 5 clear cell tumors, and 16 of 23 grade 3 endometrioid tumors. In conclusion, upregulation of FAK is seen in both endometrial hyperplasia and carcinoma, implying that FAK may play an important role in endometrial carcinogenesis. FAK overexpression in endometrial carcinoma correlates with higher FIGO grade and p53 overexpression. PMID- 15536335 TI - Cytokeratin 20 confirms merkel cell metastasis to stomach. AB - Merkel cell carcinoma (MCC) is an uncommon, aggressive neuroendocrine carcinoma of the skin. Its prognosis is poor because of involvement of regional lymph nodes and metastases to distant sites such as bone, liver, lung, and brain. Very few metastases are reported to the gastrointestinal tract. Several studies have shown the utility of cytokeratin 20 to differentiate MCC from other small round blue cell tumors, particularly those with neuroendocrine differentiation. In this case report, we present an unusual metastasis to the stomach and the utility of cytokeratin 20 immunohistochemistry to confirm MCC. PMID- 15536336 TI - Expression of cellular adhesion proteins and abnormal glycoproteins in human aberrant crypt foci. AB - Aberrant crypt foci (ACFs) may be the earliest recognizable histologic precursor lesion for colon cancer. ACF may develop from a complex of events, including the development of cryptal hyperproliferation, defects in the rate of apoptosis, and abnormalities in cellular adhesion. In this study, we hypothesized that human ACF would exhibit discrete differences in cell adhesion proteins compared with normal mucosa of biologic markers associated with colon cancer. ACFs were isolated from resected colon mucosa from 45 patients undergoing surgery for colon cancer. We evaluated the protein expression of 3 biologic markers that may be related to the progression of aberrant crypt foci to tumors: carcinoembryonic antigen, E cadherin, and sialyl Tn antigen. In general, ACFs located near cancers in the right colon were more often hyperplastic than dysplastic; this was more noticeable in the left colon. Carcinoembryonic antigen expression was found to be more intense in apical portions of ACF crypts, with sialyl Tn antigen moderately increased, whereas E-cadherin diffusely stained throughout crypts within ACFs. There are significant biologic changes in potential tumor markers that accompany the early transformation of the normal glandular epithelium, some of which are expressed very early in the colon at the stage of appearance of ACF. PMID- 15536337 TI - Histiocytic subpopulations in the gastrointestinal tract: distribution and possible relationship to function. AB - The distribution of specific histiocyte subsets within the human gastrointestinal tract has not been extensively characterized. Our goal was to immunohistochemically evaluate the distribution and location of CD1a-positive, CD68-positive, and Factor XIIIa (FXIIIa)-positive histiocyte subsets within the normal gastrointestinal tract and attempt to relate distribution to possible function. Twenty-nine samples of normal esophagus, stomach, small bowel, large bowel, and anus were routinely processed and immunohistochemically stained with antibodies to CD68, CD1a, and FXIIIa. The distribution and histologic location of histiocyte subsets were qualitatively analyzed. CD1a-positive cells were seen exclusively within anal and esophageal squamous mucosa. CD68 positive histiocytes were present in lamina propria and submucosa throughout the gastrointestinal tract and in Peyer patches. FXIIIa-positive histiocytes were also abundant in lamina propria and submucosa throughout the gastrointestinal tract, particularly around pericryptal sheaths and in parafollicular regions surrounding Peyer patches. Our results showed that there are distinct subpopulations of gastrointestinal histiocytes, and that distribution varies according to both cell type and site. Because Langerhans cells are epidermal antigen processing/presentation cells, their exclusive presence in squamous mucosa suggests an analogous function there. The prominence of both CD68 and FXIIIa positive cells surrounding glandular pericryptal sheaths suggests that they are important to immune function at this mucosal interface and may play a role in communication between glands and lamina propria. In addition, the presence of specific histiocyte subsets within Peyer patches and para-follicular regions suggests that they are involved in different aspects of antigen processing associated with gut lymphoid tissue. Further studies are needed to explore the relation between specific histiocyte subsets and gastrointestinal disease processes. PMID- 15536338 TI - Immunohistochemical recognition of the epidermal growth factor receptor by the h R3 antibody in the skin of experimental animals. AB - The h-R3 is a humanized monoclonal antibody (Mab) that binds to the external domain of the human epidermal growth factor receptor (EGF-R). It has being used for the treatment of head and neck tumors. Since it is known that different animal species have different EGF-R amino acid sequences, it raises the handicap that, a priori, the animal species relevant for the pharmacodynamic, pharmacokinetic, and toxicologic studies of this Mab are unknown. To elucidate relevant laboratory animal species, the authors investigated the immunohistochemical recognition by h-R3 Mab of EGF-R in small skin biopsy samples obtained from NMRI nu/nu, C57Bl/6, and OF-1 mice; Sprague-Dawley rats; Hartley guinea pigs; New Zealand rabbits; and Cercopithecus aethiops and Macaca fascicularis monkeys. Additionally, three human skin biopsies were obtained from trauma victims. The immunolocalization of EGF-R in different tissue sections was performed using the avidin-biotin peroxidase complex (ABC) technique. The skin sections from different tissues were tested with the biotinylated h-R3 Mab or with a biotinylated irrelevant Mab, used as negative control. The staining intensity was qualified as:-, no staining; +, weak staining; ++, moderate staining; +++, strong staining. Macaca fascicularis monkey, New Zealand rabbit, and OF-1 mouse skins had a strong staining intensity; Cercopithecus aethiops monkey and Sprague-Dawley rat skins showed a moderate to strong staining intensity; C57Bl/6 mouse skins showed a moderate staining intensity. No staining was observed in the skins from Hartley guinea pigs and MNRI nu/nu mice. The fact that h-R3 Mab recognizes other animal EGF receptors in addition to the human EGF R makes it possible to perform relevant pharmacodynamic, pharmacokinetic, and toxicologic studies in some laboratory animals. PMID- 15536339 TI - Immunohistochemical analysis of tyrosine phosphorylation and AP-1 transcription factors c-Jun, Jun D, and Fos family during early ovarian follicle development in the mouse. AB - The growth control mechanism of early-stage ovarian follicles is unknown. Tyrosine phosphorylation of signaling molecules and changes in expression and activation of AP-1 transcription factors have been implicated in growth regulation of numerous cell types. In this study, we used immunohistochemistry to analyze tyrosine phosphorylation patterns and expression and activation of selected AP-1 transcription factors in mouse ovarian follicles. The ovaries were collected from B62F1/J mice in estrus. Representative sections were immunostained for phosphotyrosine, phospho-c-Jun, Jun D, and c-Fos. Phosphotyrosine staining was perioocytic from the transitional stage until approximately 5 to 7 layers of granulosa cells had formed. Perioocytic staining was then replaced by scattered stippled staining in granulosa cells of larger follicles. Phospho c-Jun was exclusively expressed in mitotic granulosa cells of follicles from transitional to antral stages. Jun D was expressed in the oocytes of primordial, primary, or transitional follicles and disappeared at the 2-layer preantral stage. Fos was present in corpora lutea and theca cells but not in granulosa cells. Collectively, these data indicate that phosphotyrosine signaling and AP-1 transcription factors are intimately involved in early stages of ovarian follicle growth. PMID- 15536340 TI - An easy method for manual construction of high-density tissue arrays. AB - Tissue array technique is a powerful tool for high-throughput in situ analysis on a large cohort of cases. This report describes an easy and effective method for manual construction of high-density tissue array containing 88 (11 x 8) samples, each of which measures 2 mm in diameter. No extraneous device is needed except a conventional 16-gauge bone marrow biopsy trephine apparatus to puncture the paraffin blocks. The authors constructed 563 cases of epithelial neoplasm into 7 blocks. The sectioning is smooth and does not require adhesive tape. They performed immunohistochemical staining for cytokeratins 7 and 20 on these tissue array slides. The samples rarely fell off during the antigen retrieval and staining procedure. The results were generally in agreement with those in previous reports. The authors offer a satisfactory alternative method for building custom arrays for any laboratory that is unable to afford a tissue array apparatus. PMID- 15536341 TI - Assessment of T-cell clonality via T-cell receptor-gamma rearrangements in cutaneous T-cell-dominant infiltrates using polymerase chain reaction and single stranded DNA conformational polymorphism assay. AB - Discerning the pathologic significance of cutaneous T-cell infiltrates can pose a diagnostic challenge for dermatopathologists. Reactive conditions such as drug associated lymphomatoid hypersensitivity and lymphomatoid lupus erythematosus can demonstrate lymphoid atypia and a phenotype resembling cutaneous T-cell lymphoma (CTCL). Further, lymphoid dyscrasias such as pityriasis lichenoides chronica, large plaque parapsoriasis, and atypical pigmentary purpura confuse the picture because they not only mimic CTCL but also represent prelymphomatous states with inherent malignant potential. Although the emergence of a dominant clone has been considered a clue indicative of a T-cell dyscrasia, there are reports concerning the identification of monoclonality in biopsies of reactive lymphoid infiltrates. We have conducted a modified single-stranded DNA conformational polymorphism (SSCP) assay using paraffin-embedded, formalin-fixed tissue on 92 T-cell-rich biopsies to determine the relative specificity and sensitivity of this methodology. In addition, laser capture microdissection (LCM) was performed on 22 of the 92 samples to isolate the area of interest and to compare its specificity and sensitivity with those SSCP assays performed without LCM. We found that monoclonality or oligoclonality is 86% specific for preneoplastic and neoplastic states, whereas the finding of polyclonality appears to be relatively specific for a reactive process. Some cases of reversible T-cell dyscrasia produced a molecular profile mimicking lymphoma or prelymphomatous states by virtue of monoclonality or oligoclonality. Although LCM appears to improve the sensitivity for detecting preneoplastic conditions, the relative specificity appears to be the same as that encountered with routine SSCP. PMID- 15536343 TI - Primary meningeal Epstein-Barr virus-related leiomyosarcoma in a man infected with human immunodeficiency virus: review of literature, emphasizing the differential diagnosis and pathogenesis. AB - We describe the clinical, radiologic, surgical, and pathologic findings of a 29 year-old Peruvian human immunodeficiency virus-infected man with a primary parasellar meningeal leiomyosarcoma involving the left lesser esphenoidal wing and the cavernous sinus. Over a period of 13 months, he developed headache, vomiting, insomnia, and diplopia. Magnetic resonance imaging revealed a left parasellar extra-axial mass that was isointense in T1, hypointense in T2, and gadolinium-enhanced. The patient underwent subtotal resection of the tumor. The neoplasm was composed of spindle cells with smooth-muscle features. It showed moderate atypia, inconspicuous nucleoli, and scanty mitosis. No tumor necrosis was detected. The immunohistochemistry revealed strong positivity for vimentin, desmin, and smooth-muscle alpha-actin. A low-grade leiomyosarcoma was diagnosed. The in situ hybridization showed positive nuclear reactivity for Epstein-Barr virus-encoded RNA. The immunohistochemistry was negative for Epstein-Barr virus latent membrane protein 1. The main differential diagnosis of primary meningeal smooth-muscle tumors includes meningioma and peripheral nerve sheath tumors. Epstein-Barr virus has been demonstrated in most smooth-muscle tumors associated with acquired immune deficiency syndrome (AIDS). Primary meningeal smooth-muscle tumors, exceedingly rare neoplasms, remarkably affect young adults with AIDS. Comparatively, most AIDS-related visceral (nonmeningeal) smooth-muscle tumors have been reported in children. The permissiveness and tumorigenesis associated with Epstein-Barr virus may depend on the age of human immunodeficiency virus infection. PMID- 15536342 TI - p53 alterations in colon tumors: a comparison of SSCP/sequencing and immunohistochemistry. AB - This study compares single-strand conformation polymorphism (SSCP)/sequencing and immunohistochemistry (IHC) in a population-based colon cancer study to determine the best methods to evaluate p 53 alterations in tumors. Epidemiologic data collected from the Utah portion of a multicenter case-control study of colon cancer (n = 268) was used to compare somatic p53 mutations detected using SSCP/sequencing of exons 5 through 8 with those with p53 protein overexpression detected by IHC. A total of 136 tumors (51%) had p 53 mutations identified using SSCP/sequencing. IHC detected 164 tumors (61%) with protein overexpression (using a cut point of > or =20% positive cells) and 142 tumors (53%) when > or =50% positive cells were used. Sensitivity of IHC (> or =20% level) using SSCP/sequencing as the reference method was 85%. Specificity of IHC (> or =20% level) using SSCP/sequencing as reference was 63%. When > or =50% positive cells were used, specificity increased to 77%. Associations with age, gender, tumor site, stage, and Ki-ras were similar for both methods. An inverse relationship between microsatellite instability and p 53 was detected with the higher threshold for IHC positivity and SSCP/sequencing. SSCP/sequencing was able to discriminate between mutated p 53 and wild-type p 53 when evaluating dietary associations whereas IHC was not able to discriminate between these tumor types. Using a level of 50% or more positive cells increases specificity relative to sensitivity in comparison with lower staining levels, and is comparable with sequencing in its ability to detect an inverse relationship with the MSI. Advantages gained by sequencing are its ability to examine specific mutations and the improved ability to discriminate between cases with p 53 mutation and wild type when evaluating associations. PMID- 15536345 TI - [Occupational lung cancers]. AB - Bronchopulmonary cancers are the most prevalent of all occupational cancers. Many work-involved cancerous agents have been identified, the first in line being asbestos. These cancers have no clinical, radiological or histological specificity. Only a standardised, systematic enquiry in all the patients suffering from lung cancers will reveal an eventual occupational exposure. Twelve tables of occupational diseases in the general regime of the national health scheme permit the declaration of an occupational disease and the compensation of patients and entitled persons. Patients exhibiting lung cancers related to the inhalation of asbestos, recognized as an occupational disease, benefit from particular dispositions. PMID- 15536346 TI - [Screening for lung cancer]. AB - Bronchial cancer is the ideal candidate for the application of screening strategies: it is a serious, frequent disease and its diagnosis is often delayed. Moreover, only the early stages of the disease offer high hopes of being cured. However, proof of the impact of screening has not been supplied either by X-ray or by cytological examination of expectorations, despite numerous open and subsequent randomised trials. Recently, the low dose scan has shown to be a technique far more sensitive than X-ray and capable of detecting early stage tumours. Nevertheless, it is also a barely specific examination revealing a great number of benign abnormalities and perhaps leading to futile examinations. Because of this, the eventual demonstration of the impact of the scan on the lung cancer mortality warrants the development of randomised trials. PMID- 15536347 TI - [18Fluoro-deoxy-glucose positron emission tomography (18FGT-PET) and lung cancer]. AB - In pulmonary oncology, positron emission tomography provides metabolic imaging with a sensitivity and specificity of around 90%. Such results are of particular interest in the diagnosis of the benigness or malignancy of a pulmonary nodule, for the staging and the follow-up of lung cancers. Studies are ongoing to assess its interest in the appreciation of the prognosis and response to chemo- and radiotherapy. PMID- 15536348 TI - [Cardiorespiratory limits to the surgical treatment of lung cancer]. AB - Cardiorespiratory co-morbidity is a predictive factor of post-surgical mortality and morbidity. In the case of lung cancer, the pre-therapeutic work-up must assess the post-surgical risks by integrating such co-morbidity. In view of this, predictive scores and decisional algorithms have been developed. However, such tools were developed and assessed only to predict post-surgical risks during the first or second month following resection. Till now, prediction of long term quality of candidates for pulmonary resection has not be studied, although the question has often been raised by the patient and the medical and surgical teams, notably in the case of cardiorespiratory limitation prior to the intervention. A study on the predictability of the quality of life at 6 months following pulmonary resection was conducted in the Tenon hospital in a cohort of 81 patients, candidates for resection, initially selected on an MMFR lesser than 80% of the reference. Out of the 43 patients who finally underwent resection and were analysed at 6 months, there were few predictive factors for the alteration in quality of life at 6 months following pulmonary resection, other than the extension of the surgical act. Conversely, this alteration did not significantly depend on the immediate post-surgical events and can be explained by the deterioration in respiratory function. PMID- 15536349 TI - [Should lymph node extension be handled by curage or sampling?]. AB - Between 10 and 20% of patients exhibiting normal mediastinum during surgery will present, on histology, invaded lymph nodes when complete curage is performed and, in 25% of cases the mediastinal lymph nodes may be invaded without involvement of the with the hilar lymph nodes. Moreover, such involvement of the mediastinum not uncommon in peripheral tumours. Hence it is clear that multiple mediastinal node samples should be taken. Should simple "sampling" be performed as is recommended by the Lung Cancer Study Group or lymph node curage? The morbidity inherent to curage and the fact that, despite several randomised studies, the impact of curage on survival has not been clearly demonstrated, explains the continued debate. PMID- 15536350 TI - [Pre- and peri-surgical chemotherapy of stage I and II resectable non-small cell lung cancers]. AB - Chemotherapy (CT) combined with surgery in non-small cell lung cancers has been studied for a number of years. It can be used prior to or following surgery (adjuvant). A long rather unfruitful period ended with the meta-analysis of the Non-Small Cell Cancer Collaborative Group, published in the British Medical Journal in 1995 that suggested an increase in survival of 5% at 5 years with the addition of adjuvant chemotherapy to surgery. Since this publication, arguments have accumulated in favour of this combination. Phase II studies have shown the feasibility of pre-surgical CT. A randomised trial in France showed a near 10% improvement in survival at 5 years, approaching statistical significance, and that this beneficial effect was further enhanced in the early stages of cancer. Excess post-surgical morbidity and mortality, even though non-significant, emphasizes the need for an effective but less toxic CT than the mitomycine ifosfamide-cisplatin combination initially selected. In the field of adjuvant CT, the arguments in favour of the association have accumulated with the positive results of 3 studies, the IALT trial, the BR10 trial of the Canadian National Cancer Institute and the 9633 trial of the CALGB, with the latter two studies presented this year at the American Society of Clinical Oncology meeting. Four other pre-surgery CT trials are ongoing, but their results will not be available for several years. When choosing optimal timing among the various CT administration methods, before or after surgery, the arguments are in favour of pre-surgery CT: the possibility of assessing the chemosensitivity of the tumor, permitting the early withdrawal of treatment if it fails (presently in 40% of patients), the enhanced acceptability of CT by the patients, and the increase in resectability of the tumours. Conversely, however, one must note the greater difficulty for staging and the increase in post-surgical risks, basically in N2 patients, which will gradually lead to its replacement by the use of 3rd generation CT. PMID- 15536351 TI - [Which treatments should be proposed in resectable stage III cancers?]. AB - The results of surgery alone or associated with an adjuvant treatment, chemo- and/or radio-therapy, in the management of stage III resectable non-small cell lung cancer (NSCLC), are disappointing; with survival rates at 5 years generally lesser than 20% and many local and distant relapses. They justified the development of phase II trials that showed the feasibility of the chemotherapy and neo-adjuvant chemo-radiotherapy with high rates of response and acceptable toxicity. The three phase III studies with induction chemotherapy gave contradicting results; two of them were on small cohorts and presented methodological biases. Regarding pre-surgery chemo-radiotherapy, there is presently no published phase II trial and a recent phase II trial was in favour of a concomitant mode with bifractioned irradiation. The new targeted therapeutic agents do not yet have an indication in this field. In the particular case of resectable tumours of the apex invading the wall, so-called Pancoast, neo adjuvant chemoradiotherapy followed by surgical resection has become the treatment of choice. The place of chemotherapy, radiotherapy and even surgery in the treatment of resectable stage III NSCLC remains to be defined and the inclusion of patients in randomised clinical trials is recommended. PMID- 15536352 TI - [Treatment of non-resectable stage III non-small cell lung cancer]. AB - The prognosis of locally advanced and non-resectable non-small cell lung cancers (NSCLC) remains poor, despite the therapeutic progress made over the past few years. The best therapeutic strategy is currently the concomitant association of chemotherapy and radiotherapy. Such treatment must be reserved for patients in general state of good health. Oesophagitis is the principle limiting toxicity. The objectives for the years to come are to improve the results and to reduce the toxicity by optimising radiotherapy, using new chemotherapy drugs, and by pinpointing the therapeutic strategy. The sequential association of chemotherapy and radiotherapy still retains its place for the other patients. PMID- 15536353 TI - [The place of interventional endoscopy in the treatment of lung cancer]. AB - Interventional endoscopy, by eliminating an obstacle compromising the survival of a patient, has its place in the treatment of bronchial cancer whatever its stage of evolution. Forty percent of patients have not been treated yet, 30 still have therapeutic possibilities at the time of their relapse and for 30% all the possible treatments have been administered and interventional endoscopy remains their only chance of survival. A strict operational protocol ensures safety: operating theatre, general anaesthesia, rigid bronchoscopy, jet-ventilation, post surgery ICU, and systematic post-surgery fibroscopy. Eighty percent of the patients retrieve normal ventilation. 12% are not sufficiently improved and 3% die either from uncontrollable haemorrhage or from the impossibility of repermeation. The nature of the tumour (primary bronchial cancer, metastasis at distance, invasion from a surrounding cancer) does not enter into the decision to intervene. The benefits of the latter are assessed by the pneumologist or oncologist who is in charge of the patient, but the final decision to intervene naturally remains with the endoscopist. PMID- 15536354 TI - [First-line treatment of metastatic non-small cell lung cancers]. AB - The first-line treatment of non-small cell bronchial cancers at the metastatic stage relies on chemotherapy, the impact of which, in terms of survival in comparison with palliative care alone, has been confirmed by several meta analyses. The recent development of new drugs (vinorelbine, gemcitabine, paclitaxel, docetaxel) that benefit from an improved efficacy-toxicity ratio, has led to the individualisation, as treatment standard in patients in a general state of good health, of bitherapies based on the combination of a platinum salt and recent active drugs. The latter exhibit similar efficacy, in terms of response and survival, providing PS 0 or 1 patients with a median survival of around 8 months and, at 1 year, nearing 35%. The choice of the first-line therapy in a given patient will be based essentially on the differences in toxicity profiles among the therapeutic regimens, taking into account the frequent co morbidities in this population, the modalities of administration and the perspectives of second-line therapy. The patients with an altered general state of health (PS 2) will benefit from adapted treatment based on active monotherapy or bitherapy without cisplatin. The prolongation of the first-line therapy over and above 4 cycles following maximal reduction of the tumour does not provide enhanced survival and considerably increases toxicity. The recent cytotoxic agents appear to provide a real progress, although modest in the treatment of metastatic NSCBC, progress which is visible considering the proportion of patients still living in the long term. PMID- 15536355 TI - [Second-line chemotherapy in non-small cell lung cancer]. AB - The progress in first-line chemotherapy in non-small cell lung cancer with efficient and often well-tolerated combinations has progressively led to the proposal of second-line chemotherapy for the patients. Docetaxel provides significantly enhanced survival compared with palliative treatments. Various phase II studies with vinorelbine, gemcitabine and paclitaxel have been reported. Pemetrexed has demonstrated identical enhanced survival as docetaxel, but with improved tolerance. An epothilone B analog is under study in clinical trials. PMID- 15536356 TI - [Do combinations without cisplatin represent an alternative to conventional chemotherapy in non-small cell lung cancers?]. AB - Chemotherapy of advanced non-small cell lung cancers is based on cisplatin, which prolongs survival. However, the high toxicity of double-type combinations including cisplatin warrants the search for alternatives. New anti-cancer cytotoxic agents exhibit improved efficacy-toxicity ratio. Bitherapy, without cisplatin and with new molecules, provides hope of improvement in survival without deterioration in quality of life. Several randomised trials, already published or presented with mature results, suggest that certain bitherapies with new cytostatic agents are an alternative to the classical cisplatin-based bitherapies. The recent modifications in the guidelines of the American Association of Clinical Oncology take into account these recent results by admitting that "chemotherapy without cisplatin can be used as an alternative to the first-line platin-based chemotherapies". This brief review of the literature underlines the methodological questions raised by the publication of randomised studies essentially assessing gemcitabine-vinorelbine or gemcitabine-taxane combinations, by confronting them with either mono-chemotherapies or reference chemotherapy based on cisplatin. The assessment criteria selected in such studies are discussed. PMID- 15536357 TI - [The place of targeted treatments: receptor tyrosine kinase inhibitors]. AB - Targeted biological treatments are aimed at correcting one or several abnormalities inherent to cancer cells by specifically affecting one or several identified abnormal cell mechanisms. Many signalisation pathways involve membrane receptors of the tyrosine kinase receptor family, notably the HER receptors (with, first-line, EGFR or epidermal growth factor receptor) and the VEGF receptors. These receptors can be blocked either by a monoclonal antibody directed against the ligand (i.e., bevacizumab or anti-VEGF Avastin) or against the extra-cellular segment of the receptor (i.e., cetuximab or anti-EGFR Erbitux) or by tyrosine kinase inhibitors of the intracellular segment of the receptor (i.e., gefitinib or Iressa and erlotinib or Tarceva). The preliminary studies with these new treatments in pulmonary oncology are presented. PMID- 15536358 TI - [Methodology for the assessment of new targeted treatments in clinical trials. Future perspectives]. AB - Because targeted treatments with cytostatic effect do not usually lead to the reduction in tumoral mass, assessment of their efficacy in terms of response should differ from that of chemotherapy. The notion of "controlling the disease" and the "duration of clinical benefits" have recently been introduced and englobe response and stability. These criteria, which have not been officially validated, are frequently found, notably in studies on metastatic non-small cell lung cancers. The new targeted treatments therefore provide the occasion for us, clinicians and assisted by statisticians, to review our strategy and to progress in the construction of adapted clinical trials. PMID- 15536359 TI - [Malignant germinal tumours of the mediastinum: diagnosis and treatment]. AB - Mediastinal germinal tumours are composed of tissues resembling those that follow one another during embryo development, by differentiation of the primordial and extraembryonic layers. Such practice separates the mature teratomas (benign), seminomas and non-seminomatous germinal tumours (NSGT). Platin-based chemotherapy has shattered the prognosis of such tumours. PMID- 15536360 TI - [Which chemotherapy for small cell lung cancers?]. AB - Small cell lung cancers were a clinical entity of severe prognosis until the nineteen eighties, time at which their frequent chemosensitivity was demonstrated. Around 90% of objective responses, in the case of complete response, lead to remission. Although still rare, remission depends on the ability of the treatment administered rather than on the extent of the disease itself. The use of high-dose polychemotherapeutic protocols is a fundamental requirement but exposes the patient to limiting toxicity, notably haematological. The duration of treatments should be shortened to around six cycles in the case of complete response with, whenever possible, closer periodicity. Such efficient chemotherapy can only be applied in the context of a wider strategy, notably in combination with other therapeutic means: thoracic radiotherapy of the localised forms, curative or prophylactic cranial irradiation and adjuvant medical treatments. In the case of relapse, second-line chemotherapies are proposed but their efficacy is lesser and limited in time. PMID- 15536361 TI - [Modalities of radiotherapy in small cell lung cancer: thoracic radiotherapy and prophylactic cerebral irradiation]. AB - Small cell lung cancers (SCC) represent 20% of all lung cancers. After the initial control of the extension, only one third of the patients with SCC will finally have limited disease. The treatment of limited SCC currently relies on chemo-radiotherapeutic combinations that have improved overall survival and survival without metastases over the last few years. Nevertheless, even in limited forms, survival at 5 years varies from 10 to 15% and rarely exceeds 25% in the best series. The risk of relapse is high: although around 70% of patients with a limited form will have complete response, only 15 to 20% of them will exhibit prolonged survival. Indeed, most patients relapse, and the risk of cerebral dissemination for example is particularly high, reaching 50% at 2 years even in complete responders. After the results of a meta-analysis evaluating prophylactic cranial irradiation (PCI) among SCC complete responders, demonstrating 5% enhancement of survival at 3 years, PCI is part of the standard management of SCC in complete response. Despite the improvement in overall survival with the combined treatments, the mediocre results observed in terms of long-term survival warrant further clinical trials in order to define the optimal polychemotherapeutic and radiotherapeutic modalities, the best means of combining these two therapies and the place for new therapies. PMID- 15536362 TI - [The place for therapeutic intensification in small cell lung cancer]. AB - Therapeutic intensification of small cell lung cancer (SCLC) relies on either the simultaneous and repeated administration of a greater number of drugs without cross-resistance, or to the use of classical combinations with greater intensity, either by increasing the doses in each cycle or by reducing the intervals between cycles. The authors review the trials concerning disseminated SCLC: eight randomised studies have compared a standard regimen with an intensified regimen: only one has demonstrated a benefit in terms of survival with the intensified regimen. They will then discuss the problem of intensification of the chemotherapy in the particular case of localised SCLC, which requires the combination of chemotherapy and thoracic radiotherapy: two studies were in favour of the intensified regimen. Lastly, intensification, associating massive doses of polychemotherapy with autologous bone marrow graft or re-injection of hematopoietic stem cells, increases the response rate at the price of a significant increase in toxicity and costs, without benefit in terms of survival. PMID- 15536364 TI - [Nice lung, good lung? The morphometric basis of lung function]. PMID- 15536363 TI - [What are the questions concerning small cell lung cancer? How should research be orientated?]. AB - Spectacular therapeutic progress for small cell lung cancer was obtained some thirty years ago with the introduction of polychemotherapy. Since when, except that which concerns radio-chemotherapy in the limited forms, the development of treatments has been relatively poor. Based on the analysis of published literature, the aim of this review is to discuss the research routes. The problems can be summarised in eleven principle axes: identification of new active drugs, development of new combinations with new chemotherapeutic agents, the place of analogs, the role of thoracic surgery, optimal radio-chemotherapeutic regimens, intensity of chemotherapy, the place of haematological growth factors, the duration of chemotherapy and maintenance treatments, biological treatments, prevention and treatment of cerebral metastases and salvage chemotherapy. PMID- 15536365 TI - [Tobacco smoking: world scourge, worldly answer]. PMID- 15536366 TI - [Home-based pulmonary rehabilitation in the 21st century]. PMID- 15536367 TI - [Chronic cough in children]. PMID- 15536368 TI - [Prevention of COPD exacerbations: conducting a clinical trial evaluating the benefits of inhaled steroids]. PMID- 15536369 TI - [Mandibular advancement devices for obstructive sleep apneas: need for the cooperation of specialists]. PMID- 15536370 TI - [Tobacco consumption of students ages 8 to 22 in Abidjan, 2002]. AB - BACKGROUND: To update full time educated youth data on smoking prevalence related to gender and to assess perception and behaviour related to smoking, we have led a self-administrated question-study from January to June 2002 in Abidjan with students from 8 to 22 years old. METHODS: Our population was divided in 3 groups: T1 (812 years), T2 (13-17 years) and T3 (18-22 years). The size of each group has been determined using the smoking estimated rate in each of them. RESULTS: 2742 students had returned a well-full questionnaire, with the following rates, expressed by mean and standard deviation in%: a total smoking rate at 7.9 +/- 0.5, with statistic difference between boys and girls (11 +/- 0,8 versus 3,7 +/- 0,5; p<0.001). This rate increased with age: 3.7 +/- 0.8 in T1; 12.9 +/- 1.0 in T2 and 17.1 +/- 1,1% in T3. Globally, the rate was 0.7 +/- 0.2 for regular smokers (>or=1 cigarette/day) (10% of C.D.T.) and 1.7 +/- 0.2% for ex-smokers. 99.2% of current smokers used cigarettes only. On average, the tobacco consumption was 3.4 cigarettes/day. In non-smokers group, 88% had respiratory symptoms linked to smoke exposure, with 17% of major dyspnea. Non smokers reported public places as the most frequent (66.7 +/- 1.3%) and the highest long time exposure (44.3 +/- 1.9%) places to second hand smoke. DISCUSSION: A similar smoking prevalence between asthmatics and non-asthmatics subjects, and between sporty type and non sporty type subjects, suggested that students had superficial knowledge or under assessment on smoking detrimental effect. However, their perception of smoking health hazard was sufficient to give 95 +/- 1% favourable opinions on necessary smoking place regulations and 85 +/- 1% favourable opinions on tobacco product advertising ban. CONCLUSION: The epidemiological and behaviour information given by this study might contribute to the development of a national youth tobacco control program, provided a complementary national survey would be led including non educated youth. PMID- 15536371 TI - [Usefulness of biological markers in the evaluation of smoking at the first visit of a smoking cessation program]. AB - INTRODUCTION: We have studied the relationship between urinary cotinine and other tobacco markers to assess the relevance of urinary cotinine in the dosage of nicotine substitutes. METHODS: One hundred and fifty-five patients, who were regular cigarette smokers not taking any nicotine substitutes took part in the study. The parameters examined were daily consumption, nicotine contents of the cigarettes, the Fagerstrom test score, levels of carbon monoxide (CO) in exhaled breath air and urinary cotinine. RESULTS: The most significant correlation was between urinary cotinine and exhaled CO (Spearman rs=0,59 p<0,001). The correlation between urinary cotinine, the Fagerstrom test (rs=0,32 p<0,001) and daily cigarette consumption (rs=0,32 p<0,001) was less significant. The nicotine content was not related to urinary cotinine (rs =-0,10NS). CONCLUSIONS: For economic reasons measuring urinary cotinine is not necessary when the CO level is or=35 ppm. It becomes useful if the CO level is between 15 and 34 ppm, as CO level in itself is insufficient to determine the dosage of nicotine substitutes. PMID- 15536372 TI - [Home pulmonary rehabilitation: results in a cohort of 37 patients with respiratory handicap]. AB - INTRODUCTION: The purpose of this open, prospective, non-controlled study is to evaluate the feasibility and effect of 10 weeks of home-based exercise retraining on a cyclo-ergometer in respiratory patients. METHODS: 75 patients are consecutively referred. 37 patients, (75% COPD), took part in the programme, 38 (51%) did not. Nine dropped out, five had missing data, therefore the analysis is of 23 patients. RESULTS: There was a significant improvement in Quality of life (CRQ) (p=0,02), in endurance and in work rate on the bicycle (p<0.001). There is a correlation between the increase in work rate on the bicycle and the improvement in quality of life (p=0.01, r=0.5). When the patients are divided according to the level of improvement in work rate on the cycle-ergometer (mild 0 to 10% improvement; moderate 11 to 49% and strong > 50%) we find that only mild improvement is found in those with a low initial work rate. CONCLUSION: We confirm the effectiveness of home based training, this being feasible in one out of two patients referred. Co-morbidity and the psycho-social situation were the main exclusion factors. The improvement in effort capacity could be an important determinant of quality of life. Cyclo-ergometer could required too great an effort for training more severe patients. PMID- 15536373 TI - [Evaluation of advanced non-small-cell lung cancer (ANSCLC) chemotherapy in routine practice]. AB - BACKGROUND: Patients in ANCCLC chemotherapy-trials are not representative of the general lung cancer population. The purpose of this study was to describe the ANCCLC population routinely treated by chemotherapy in a general hospital, and to assess the results of chemotherapy in this population. METHODS: All newly diagnosed IIIB/IV ANCCLC chemotherapy-treated patients over a three-year period were prospectively assessed for response rate, toxicity and survival. RESULTS: Seventy seven patients (70% stage IV, 69% PS 1/0, 30% with cerebral metastases, 60% ineligible for major lung trials) received first-line chemotherapy (cisplatine or paraplatine with vinorelbine) with tumor control in 31 (40%) and symptom improvement in 20 (26%) patients. 17 (22%) patients experienced febrile neutropenia. 33 (43%) patients received second line chemotherapy (gemcitabine) with tumor control in 12 (36%) and symptom improvement in 9 (27%) patients. Overall median survival was 7 months and 30% patients were alive at 1 year. CONCLUSIONS: Routinely ANCCLC chemotherapy-treated patients in our center have poor prognostic factors and many comorbidities. Chemotherapy results in tumor control in 43% patients, - of whom two thirds have symptom improvement -, with a high rate of febrile neutropenia. PMID- 15536374 TI - [Home-based pulmonary rehabilitation: the evidence, practical issues and medico economics]. AB - INTRODUCTION: Pulmonary rehabilitation is an established part of the management of chronic obstructive airway disease. For longer-term effects, extending rehabilitation into domiciliary care settings may be necessary. There are few studies evaluating precisely this modality of management in a home setting. STATE OF THE ART: This review analyses the current literature on home based rehabilitation. The benefits of pulmonary rehabilitation, the practical aspects of its application, and its advantages and limitations are detailed. The few data on economic aspects of home rehabilitation are also discussed. PERSPECTIVES: Home based pulmonary rehabilitation is effective with positive short-term effects on quality of life, breathlessness and effort tolerance. Its practical application needs to be defined. CONCLUSION: Home-based pulmonary rehabilitation appears to be an effective intervention in patients with COPD but more studies are necessary to evaluate it fully. PMID- 15536375 TI - [The role of the small airways in childhood asthma]. AB - INTRODUCTION: Asthma, the most common chronic disease in paediatrics, is characterised by an inflammatory process involving the proximal and distal airways, with remodelling of the airways and changes in respiratory function. STATE OF THE ART: Recent studies in children have provided evidence of damage to the small airways identical to that occurring in adults. In fact inflammatory cells as well as structural alterations of the airways have been found in both bronchial biopsies and broncho-alveolar lavage fluid in infants and young children. At the functional level this damage is manifest by a reduction in maximum expiratory flow rates between 25-75% of the vital capacity. It also shows itself by the demonstration of thick walled bronchi and hypodense bands on the high resolution CT scan. It appears that conventional inhaled treatments have no effect on the small airways. Finally the main prognostic factors for this distal damage are respiratory infections and exposure to tobacco in utero. PERSPECTIVES: Damage to the small airways in childhood asthma indicates the need for new therapies aimed at this part of the respiratory system. CONCLUSION: Inflammation and remodelling of the small airways develops early in an asthmatic child whose respiratory system is growing rapidly and persists throughout life. PMID- 15536376 TI - [Chronic cough in childhood]. AB - INTRODUCTION: Although less frequent than viral induced recurrent cough; chronic cough remains a sometimes difficult to resolve diagnostic tool. STATE OF THE ART: Most authors estimate that a cough can by considered as chronic after three weeks of duration. Few papers have been published concerning etiologic diagnosis of chronic cough in childhood but these indicate the same main causes as in adults: cough variant asthma, postnasal drip syndrome, gastro-esophageal reflux. Nevertheless, each age bracket presents specific diagnosis: malformations between zero and one year, psychogenic cough in adolescents. PERSPECTIVES: New techniques as induced sputum studies helps to refine chronic cough diagnosis in childhood (after 7 years). Eosinophilic bronchitis, associated or not to bronchial hyperresponsiveness has important therapeutic consequences because associated with a favourable response to corticosteroids. Other techniques will be developed in the future (exhaled NO for example). CONCLUSIONS: Chronic cough in childhood must be investigated from an anatomic point of view and on frequency arguments. Control and removal of the cough will only be obtained if a precise diagnosis and a suitable treatment are reached. PMID- 15536377 TI - [Mechanisms of chronic cough pathophysiology]. AB - INTRODUCTION: In some situations such as post-virus or post whooping cough, a non productive subacute cough may occur without apparent local inflammation, epithelium abnormalities or bronchoconstriction. This subacute or chronic cough represents a real syndrome (cough disease) due to the central nervous system (CNS) and its ortho and parasympathic outputs. At the CNS level, functional disturbancies and neosynaptogenesis can be described, with the intervention of the NMDA-type glutamatergic receptors. STATE OF ART: The neurons located in the expiratory area of the breathing center (Pre-Boetzinger complex of the lower brainstem) present exagerated responses to stimuli, due to the repetitive stimulation of the NMDA receptors; this phenomenon is similar to long-term potentiation (LTP), the molecular basis of learning, memory and neosynaptogenesis. The cough reflex is thus amplified and rapidly chronic and would justify any pharmacological intervention at the NMDA-receptors level. PERSPECTIVES: More recently 5TH4 receptors have been implied in the control of respiration; an overexpression of these receptors in the Pre-Boetzinger area could contribute to an increase of the cough reflex. CONCLUSION: The present review aims at summarizing the main rationale target to pharmacologically block the chronic cough. PMID- 15536378 TI - [Clinical and immunopathological aspects of hypersensitivity pneumonitis]. AB - INTRODUCTION: Hypersensitivity pneumonitis (HP) is a pulmonary disease with symptoms of dyspnoea and cough resulting from the inhalation of an antigen to which the patient has been previously sensitized. STATE OF ART: Acute and subacute HP represent the most active forms of the disease which may become chronic while remaining progressive. HP may also evolve to end-stage lung disease. Clinical symptoms and signs tend to be non-specific and the diagnosis of HP often relies on the clinical context. The immune response is initiated when the alveolar macrophage phagocytoses the antigen, provoking the expansion of lymphocytes T and B that reach the pulmonary parenchyma through the systemic circulation. This reaction is amplified by the expression of a number of inflammatory mediators. PERSPECTIVE AND CONCLUSION: This article summarizes our current understanding of the diagnostic approach and immunological mechanisms related to HP. PMID- 15536379 TI - [Initiation and management of home mechanical ventilation]. AB - INTRODUCTION: Effective initiation and optimal monitoring of treatment are essential elements for successful home mechanical ventilation (HMV). STATE OF KNOWLEDGE: Ventilation is best initiated in a hospital setting. There is an enormous range of equipment available but there is little evidence pointing to the superiority of one device over another. Many problems can arise that lead to an interruption in HMV but often these can be resolved simply as discussed in this article. PERSPECTIVES: Unfortunately a number of pitfalls in management remain, the main one being the absence of accurate data about the sleep of patients on ventilators, which hinders optimal nocturnal management. Another problem is the lack of adaptation of equipment for use by handicapped patients. Finally the transfer of responsibility and workload to home carers including family members with disengagement by the hospital can also be a major milestone. CONCLUSION: Successful initiation and supervision is the key to effective home mechanical ventilation. Treatment is often abandoned because of pitfalls, mistakes and lack of knowledge. This article proposes means to improve these two important areas. PMID- 15536380 TI - [The COPD-PE study: prevalence and prediction of pulmonary embolism in acute exacerbations of chronic obstructive pulmonary disease]. AB - INTRODUCTION: Patients suffering from chronic obstructive pulmonary disease (COPD) are often admitted to hospital with an exacerbation of their disease that manifests itself with an increase in cough and/or sputum, increased dyspnoea and sometimes chest pain. These episodes are most often attributed to an acute exacerbation of bronchial infection but the real cause is often unknown. COPD is considered a risk factor for pulmonary embolism (PE) and PE is part of the differential diagnosis of an acute exacerbation of COPD. However, the symptoms of these two conditions overlap to a considerable extent and the investigation of PE is often ignored in these patients. Therefore the true prevalence of PE in this situation is unknown. Nevertheless several small series suggest that up to 30% of patients seen as emergencies with exacerbations of COPD may have a deep vein thrombosis or PE. The aims of this study are therefore: 1. To evaluate the prevalence of PE in patients admitted as emergencies with exacerbations of COPD; and 2. To attempt to develop a prediction guide for PE in this population in order to identify the patients in whom PE should be investigated. MATERIALS AND METHODS: 600 consecutive patients admitted as emergencies with exacerbations of COPD will be included in a cross sectional study in three university hospitals (Geneva and Lausanne in Switzerland, and Amiens in France). Patients fulfilling the inclusion criteria will be investigated by a diagnostic algorithm appropriate for the study of PE, including D-dimer levels and, in those with abnormal results, ultrasonic scan of the deep veins of the legs and spiral CT scan. The patient characteristics on admission will be incorporated in a multivariate regression analysis in an attempt to identify the predictive factors for PE in these patients. The expected duration of the study is 24 months. EXPECTED RESULTS: This study should determine the prevalence of PE in patients admitted as emergencies with exacerbations of COPD and therefore help decide when, and in which patients, a systematic search for PE should be undertaken. PMID- 15536381 TI - [Analysis of variance, part 1 (the t-test and ANOVA)]. PMID- 15536382 TI - [Analysis of variance, part 2 (linear regression)]. PMID- 15536383 TI - [Sleep apnea syndrome after total laryngectomy]. AB - INTRODUCTION: Tracheostomy was the treatment of choice for sleep apnoea syndrome before nasal continuous positive airway pressure was developed. The occurrence of sleep apnea syndrome after total laryngectomy is unusual. CASE REPORT: The authors report the case of a 73 year-old man presenting with the symptomatology of sleep apnoea syndrome, several years after a total laryngectomy for cancer. The polysomnographic recording confirmed severe sleep apnoea syndrome with an apnea-hypopnea index of 89. Clinical examination showed a marked reduction in the diameter of the tracheostomy orifice during forced inspiration whilst lying supine. The combination of negative airway pressure and muscle hypotonia during sleep could explain this closure. Use of a silver tracheostomy cannula reversed the symptoms and normalised the apnea-hypopnea index. CONCLUSION: Sleep apnea syndrome is an unusual complication of total laryngectomy, resulting from collapse of the tracheostomy walls during sleep, which can be treated by the insertion of a silver tracheostomy tube. PMID- 15536384 TI - [Use of talc and sarcoidosis - pathogenic role of cutaneous talc exposure in sarcoidosis]. AB - INTRODUCTION: We report on two patients with sarcoidosis with disseminated nodes, who used talc on irritated cutaneous areas. CASE REPORT: A histologic examination with intense polarised light showed up cristalline bi-refringent particles within vessels in contact with granulomatous areas. Microdissection followed by an electronic microscopy study and microanalysis was realised. In situ microanalysis allowed us to identify bi-refringent particles with a size of roughly 0.25microm as silica or silicate coming possibly from talc. We consequently studied a brand name talc. The diffraction spectrum showed that this product not only contained talc but also chlorite and quartz. Electron microscopy examination showed particles of all sizes even smaller than 0.25microm. These infra-microscopic particles, visible in a vessel only when agglomerated, could be invisible under optic microscopy (resolution: roughly 0.5microm) inside the granuloma even though they are responsible for it. Moreover, at this level of size of particles, they may escape mineralogic analyses which use methods involving the destruction of organic material, the mineral residue collecting on cellulose filter with a diameter generally of 0.45microm. CONCLUSION: Two recent epidemiologic studies confirm the possible role of mineral exposure in sarcoidosis. Some sarcoidosis could be caused by mineral overload on genetically predisposed patients. Some cases could be related to mineral powder application. Among different types of mineral exposure, applications of cosmetic products may induce disseminated granulomatous reaction on genetically predisposed patients. Such applications have to be considered in epidemiologic studies. PMID- 15536385 TI - [Interstitial fibrosis in cryptogenic organizing pneumonia could be a predictive factor of relapses]. AB - INTRODUCTION: Cryptogenic organizing pneumonia (COP) is a clinicopathologic syndrome characterized by a good prognosis with steroid therapy, but frequent relapses when the dose of corticosteroid is reduced or stopped. CASE REPORT: The purpose of this study was to identify histopathologic features related to the relapse of COP. Six cases of COP that had been diagnosed using open lung biopsy were selected for evaluation. The 6 cases were put into two groups composed of 3 patients who relapsed and 3 who did not relapse. Their pathologic features were examined and compared. CONCLUSIONS: Interstitial fibrosis of the lung parenchyma could correspond to histopathologic characteristics of relapses in COP. PMID- 15536386 TI - [Renal AA amyloidosis secondary to bronchiectasis: a report of two cases (including one with Mounier-Kuhn syndrome)]. AB - INTRODUCTION: We describe 2 patients with AA renal amyloidosis secondary to bronchiectasis (one patient had tracheobronchomegaly or Mounier-Kuhn syndrome). CASE REPORTS: The time period between the diagnosis of bronchiectasis and the development of renal amyloidosis was 40 years and 30 years respectively. Both patients evolved to end-stage renal failure requiring dialysis. CONCLUSION: Although the incidence of renal amyloidosis secondary to bronchectasis has declined, it remains a dreadful complication. PMID- 15536387 TI - [Acute eosinophilic pneumonia due to recent cigarette smoking]. PMID- 15536388 TI - [Home pulmonary rehabilitation in chronic respiratory insufficiency]. PMID- 15536389 TI - [Upper airway testing: an update]. PMID- 15536390 TI - [Respiratory muscle fatigue: an update]. PMID- 15536391 TI - [Chronic obstructive pulmonary disease. Evidence based respiratory medicine: 2nd update workshop of the SPLF]. PMID- 15536392 TI - [Biologically active compounds of edible mushrooms and their beneficial impact on health]. AB - Edible mushrooms are valuable a source of biologically active compounds. Some are used in the prophylaxis and therapy of such diseases as cancer and cardiovascular disease. Their antitumor mechanism is complex. The biologically active substances in mushrooms decrease DNA damage, reduce carcinogen concentrations and their activation, inhibit the growth of cancer cells by scavenging free radicals, stimulate the immune system, and induce tumor cell apoptosis. The stimulation of the immune system by the biologically active compounds in edible mushrooms protects against cold, flu, infections, well as AIDS by inhibition of viral replication. Mushrooms contain effective substances which decrease the LDL fraction of cholesterol in blood. They also prevent the accumulation of serum triaclyglycerols, thus decreasing the risk of developing cardiovascular disease. The therapeutic properties of mushrooms result from the specific polysaccharides, such as beta-glucans and chitosans, that are present in the fructification of fungi. PMID- 15536393 TI - [The formation of new blood vessels in coronary artery disease: where we are now]. AB - New blood vessel formation is of great importance to clinicians and researchers because of its participation in many diseases. Scientists are trying to exploit the mechanisms of vasculogenesis, angiogenesis, and arteriogenesis in the treatment of coronary artery disease. These mechanisms are employed to improve myocardium vascularization in patients where both pharmacological therapy and available revascularization procedures (PTCA, CABG) are not effective. In such cases, factors including vascular endothelial growth factor (VEGF), acid fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), granulocyte/macrophage colony-stimulating factor (GM-CSF), and granulocyte colony stimulating factor (G-CSF), as well as stem cells, have been applied so far. This article is a review of the current knowledge about new blood vessel formation, especially in ischemic myocardium. We have also analyzed alternative strategies of treating coronary artery disease using the mechanisms of angiogenesis, arteriogenesis, and vasculogenesis. PMID- 15536394 TI - [Involvement of apoptosis and proinflammatory cytokines in the pathogenesis of anemia in multiple myeloma]. AB - Multiple myeloma is a neoplasmatic disease of the hematopoietic system which constitutes about 10% of all hematological proliferations. Anemia is a common symptom of myeloma, especially in patients with advanced disease, and its severity correlates with the clinical stage of myeloma. There are several factors involved in the pathogenesis of anemia in multiple myeloma: infiltration of the bone marrow with monoclonal plasma cells, inadequate secretion of erythropoietin, shortened erythrocyte survival time, dysregulated iron metabolism, impaired marrow function due to proinflammatory cytokine secretion, and interaction between erythroblasts and malignant plasma cells. Recent findings indicate an important function of apoptosis in regulating physiological erythropoiesis. In physiological conditions some erythroblasts undergo apoptosis, which is induced by proteins belonging to the TNF family, i.e. Fas(CD95), FasL(CD95L),and TRAIL (TNF-related apoptosis inducing-ligand) with its receptors--DR4 (Death Receptor 4), and DR5 (Death Receptor 5). Expression of Fas, DR4, and DR5 is detected on the cell membrane of erythroblasts in all stages, whereas FasL and TRAIL are present only in more mature erythroblasts. Interaction of mature erythroblast FasL+/TRAIL+ with immature erythroblast FasL-/TRAIL--results in apoptosis of the immature cell, which contributes to the down-regulation of physiological erythropoiesis. The expression of proteins involved in erythropoiesis regulation is controlled by erythropoietin (EPO), which decreases erythroblast susceptibility to FasL and TRAIL stimulation and prevents apoptosis. On the other hand interferon gamma (IFN-gamma) and tumor necrosis factor (TNF) increase Fas expression on erythroid cells and enhance their apoptosis. Malignant plasma cells show increased expression of FasL and TRAIL and decreased expression of Fas, which make them more resistant to apoptotic signals. FasL+/TRAIL+ plasmocytes are involved in anemia pathogenesis in multiple myeloma patients by inducing apoptosis of erythroid cells. Monoclonal plasmocytes also secrete numerous cytokines involved in plasma cell growth, bone marrow neovascularisation and anemia. PMID- 15536395 TI - [Beta2-agonists--what does their chemical structure determine?]. AB - The paper presents the structure and functional relationship of beta2-agonists and the beta2-adrenoceptor. The human beta2-adrenoceptor is a member of the 7 transmembrane family of receptors. Most of the actions of the beta2-receptor are mediated through the Gs protein and the cAMP-dependent PKA system. Alternative cAMP-independent pathways affected following beta2-receptor activation have also been described. Beta2-agonists have been used as bronchodilator agents in the treatment of asthma since the development of inhaled isoprenaline preparations in 1961. Over the years, these agents have been markedly improved through the development of short-acting beta2 selective agents followed by long-acting beta2 selective agents with prolonged duration of action. Efforts directed towards improving the pharmacodynamic and pharmacokinetic properties, including the long acting and selective beta2-adrenoceptor agonists, included two major pathways of modifying the basic structure of the catecholamines, which are described in this paper. The pharmacological activity usually resides in the (R)-enantiomer. The kinetics of beta2-agonist-stimulated bronchodilation in asthma is determined by the differences in the molecular mechanism of beta2-agonists action. Regulation of the beta2-receptor is influenced by its desensitization following exposure to high concentrations of or repeated challenges with agonists, and up-regulation which can be induced by glucocorticoids and thyroid hormones. PMID- 15536396 TI - [Homocysteine--an underestimated atheromatosis risk factor. Do sex hormones influence homocysteine concentrations?]. AB - Recent data on the atherogenic effect of elevated concentrations of homocysteine (Hcy) and a review of published results on the influence of sex hormones, especially estrogens, on plasma Hcy levels are presented. Epidemiological data show that hyperhomocysteinemia as well as a mild elevation in plasma Hcy levels significantly increase cardiovascular risk. Some results do not support this relationship. Recently it was established that onset of menopause and decreases in serum estrogen levels probably increase serum Hcy concentrations. The majority of investigation, based on observational studies and the results of one randomized, double-blind placebo-controlled study, showed that use of estrogen and hormonal replacement therapy lowered HCY concentration. It has been observed that the decrease in Hcy level was greater the higher its concentrations before treatment. These results are in contrast with those of another randomized double blind placebo-controlled study which showed that the Hcy concentration does not depend on menopause and does not change after hormonal treatment. Individual data showed that dehydroepiandrosterone, an adrenal steroid, probably lowers Hcy level. There were no correlations between serum Hcy concentration and insulin concentration, body mass, and type of obesity. The hypotheses about the influence of sex hormones on Hcy concentration are not clear and need further investigation. PMID- 15536397 TI - [Potential role of the Epstein-Barr virus in the pathogenesis of systemic lupus erythematosus and kidney diseases]. AB - Epstein-Barr virus (EBV) is mainly regarded as a tumorigenic agent widely distributed in the adult population. Recent results also indicate its potential role in systemic autoimmune diseases as well as in renal disorders. This is thought to be due to the virus's capacity to invade B cells and change from the lytic linear form to the circular latent one, which allows restriction in its gene expression array. In this way the virus avoids recognition by CD4+ and CD8+ cells and escapes immunological control. Defective control of lytic and/or latent EBV infection may contribute to the development of systemic or renal pathology. PMID- 15536399 TI - The role of protease activation of inflammation in allergic respiratory diseases. AB - Extracellular endogenous proteases, as well as exogenous proteases from mites and molds, react with cell-surface receptors in the airways to generate leukocyte infiltration and to amplify the response to allergens. Stimulation leads to increased intracellular Ca ++ and gene transcription. The most thoroughly investigated receptors, protease-activated receptors (PARs), are 7-transmembrane proteins coupled to G proteins. PARs are widely distributed on the cells of the airways, where they contribute to the inflammation characteristic of allergic diseases. PAR stimulation of epithelial cells opens tight junctions, causes desquamation, and produces cytokines, chemokines, and growth factors. They degranulate eosinophils and mast cells. Proteases contract bronchial smooth muscle and cause it to proliferate. PARs also promote maturation, proliferation, and collagen production of fibroblast precursors and mature fibroblasts. PAR-2, apparently the most important of the 4 PARs that have been characterized, is increased on the epithelium of patients with asthma. Trypsin, a product of injured epithelial cells, and mast cell tryptase are potent activators of PAR-2. Mast cell chymase activates PAR-1. Proteases from mites and molds appear to act through similar receptors. They amplify IgE production to allergens, degranulate eosinophils, and can generate inflammation, even in the absence of IgE. Proteases produced by Aspergillus species to support its growth are presumably responsible for the exuberant IgE, IgG, and granulomatous response of allergic bronchopulmonary aspergillosis. Similar proteases from molds germinating on the respiratory mucosa have been recently been implicated in the pathogenesis of chronic hyperplastic rhinitis and polyps and, by extension, of intrinsic asthma. Finally, proteases from mites and fungi growing in damp, water-damaged buildings might be the basis for the increased prevalence in these buildings of rhinitis, asthma, and other respiratory diseases. Future research promises to promote our understanding of the pathogenesis of allergic respiratory diseases and point the way to new therapies. PMID- 15536403 TI - New series: Perspectives in asthma. PMID- 15536401 TI - Mechanistic actions of the risks and adverse events associated with vaccine administration. AB - Vaccine-preventable disease levels in the United States are at or near record lows. Most parents today have never seen a case of diphtheria, measles, or other once commonly encountered infectious diseases now preventable by vaccine administration. As a result, some parents wonder why their children must receive shots for diseases that do not seem to exist. Myths and misinformation about vaccine safety abound and can confuse parents who are trying to make sound decisions about their children's health care. However, we cannot take continued high immunization coverage levels for granted. A successful vaccination program, like a successful society, depends on the cooperation of every individual to ensure the good of all. This review outlines for clinical allergists immunologists the molecular basis for the risks and adverse events associated with vaccine administration so that they can be better informed as experts on vaccine-associated adverse reactions. PMID- 15536404 TI - Viral infections and asthma inception. AB - Respiratory tract infections caused by viruses have been implicated in the pathogenesis of asthma. Of these respiratory pathogens, viruses have been demonstrated to be associated with asthma epidemiologically in at least 3 ways ( Fig 1 ). First, during infancy, certain viruses have been implicated in the inception of the asthmatic phenotype. Genetic susceptibility, particularly genes coding for atopic phenotypic characteristics, might differentiate, at least in part, those children who are destined to have persistent wheezing, asthma, or both later in childhood. Second, repeated exposure to infectious viruses in daycare centers or in households with multiple older siblings increases the number of respiratory infections, but in doing so, it might paradoxically reduce the long-term risk of allergies and asthma through either pre-existing or newly formed alterations in cytokine response profiles. Third, in patients with established asthma, particularly children, viral upper respiratory tract infections play a significant role in producing acute exacerbations of airway obstruction that might result in frequent outpatient visits or in hospitalizations. This review will highlight available data on respiratory syncytial virus infections and their relationship to asthma inception in childhood. PMID- 15536405 TI - Systemic and pulmonary effects of fluticasone administered through a metered-dose inhaler in rats. AB - BACKGROUND: Metered-dose inhalers (MDIs) are convenient, simple, inexpensive, and reproducible devices for administering aerosolized drugs through the pulmonary route, but methods have not been available for use of these devices in small animals. OBJECTIVE: We sought to test the efficacy of delivery of fluticasone through an MDI to rats with a rodent-adapted spacer chamber and to compare this treatment with systemic dexamethasone for the acute pulmonary allergic inflammatory response. METHODS: Changes in body and thymus weights were used as indicators for systemic steroid effects. Rats were sensitized to ragweed pollen extract 2 weeks before the experiment, and pulmonary allergic responses were evaluated 48 hours after a single aerosolized antigen challenge on the basis of bronchoalveolar leukocytes, lung tissue sections, total lung capacity, and forced expiratory volumes. RESULTS: Inhaled fluticasone caused dose-related systemic effects, indicating successful pulmonary drug delivery. Inhaled fluticasone was more effective than placebo but less effective than systemic dexamethasone in attenuating the increase in lung eosinophils and inflammatory infiltrates and the decrease in total lung capacity associated with the allergic inflammatory response. Inhaled fluticasone prevented airway obstruction and proximal inflammation, as did dexamethasone, but it appeared to have less effect in areas of lung served by the most distal airways. CONCLUSION: This is an effective method for use of MDIs to deliver inhaled drugs to small laboratory animals, and it should be valuable for investigations of treatment effects, as well as for in vivo testing of delivery devices. PMID- 15536406 TI - Geographic variations in the effect of atopy on asthma in the European Community Respiratory Health Study. AB - BACKGROUND: Atopy has long been related to asthma. The prevalences of both atopy and asthma have shown substantial variation. OBJECTIVE: We sought to assess geographic variations in the fraction of asthma attributable to IgE sensitization to specific allergens in the European Community Respiratory Health Survey. METHODS: A cross-sectional study was undertaken during the years 1991 and 1992 on 13,558 individuals in 36 centers in 16 countries. Asthma was defined in several ways, variously incorporating reported symptoms, bronchial responsiveness to methacholine, and physician diagnosis. Specific IgE against house dust mite (Dermatophagoides pteronyssinus), cat, timothy grass, Cladosporium herbarum , and a local allergen (birch, Parietaria judaica , or ragweed) were measured. RESULTS: The overall attributable fraction (AF) of asthma symptoms caused by atopy was 30% but varied widely between centers, ranging from 4% to 61%. The overall AF increased to 43% when asthma was based on wheezing and bronchial responsiveness, to 45% with a physician diagnosis of asthma, and to 48% when the patient reported more than 12 attacks in the last year. Between centers, the AF for atopy was significantly correlated with the prevalence of atopy among the asthmatic patients ( r = 0.91) and with the sensitization to house dust mite ( r = 0.64), as well as with the prevalence of asthma among atopic individuals ( r = 0.43) and the prevalence of asthma among nonatopic individuals ( r = -0.51). CONCLUSION: The effect of atopy on the prevalence of asthma varies widely between centers, probably because of variations in factors related to the expression of asthma and to the prevalence of sensitization, particularly to house dust mite. PMID- 15536407 TI - Relationship of asthma, atopy, and bronchial responsiveness to serum eosinophil cationic proteins in early childhood. AB - BACKGROUND: The relationship between atopy, asthma, and eosinophilic inflammation is less clear in early childhood than later in life. OBJECTIVE: We sought to determine the relationships between asthma, atopy, and serum eosinophil cationic protein (ECP), a biomarker of eosinophil activation, in 6-year-old children. METHODS: Serum ECP levels were available from 968 six-year-old children who were part of a longitudinal birth cohort being assessed for asthma and atopy. Detailed clinical history and examination, lung function testing, methacholine challenge, and skin prick testing to 4 common allergens were undertaken. Subgroups of the children were compared by using t tests, ANOVA, chi 2 tests, and regression analysis. RESULTS: One hundred ninety-one (19.7%) children had current asthma, with 114 (59.7%) of these being atopic. The mean serum ECP level for the entire group was 18.0 mug/L (range, 2.0-146.0 mug/L), with no difference between male and female patients. Serum ECP was higher in atopic children (20.5 +/- 18.4), those with asthma (22.4 +/- 19.6), and those with asthma and atopy (26.6 +/- 22.4; all P < .001 compared with children with no asthma or atopy [16.1 +/- 15.9]). Serum ECP levels were highest in children with severe asthma ( P < .001), especially in those with concurrent atopy. Severity of atopy, judged on the basis of wheal size or derived variables combining wheal size and the number of positive skin tests, was a major determinant of serum ECP. Heightened methacholine responsiveness was not associated with increased serum ECP levels. CONCLUSION: The higher serum ECP levels seen in 6-year-old children with current asthma and more severe atopy suggest that atopy and eosinophilic inflammation are important in driving this clinical phenotype and that this might represent asthma that persists. PMID- 15536408 TI - Parental history of atopic disease: disease pattern and risk of pediatric atopy in offspring. AB - BACKGROUND: Family history is an important risk factor for atopic disease. However, most studies assess only limited information on family history. Because atopic disease can exhibit transient or persistent patterns, it may be useful to assess information on patterns of disease within families. This approach has been applied in other diseases, such as cancer, to discriminate between predominantly inherited versus environmentally caused (sporadic) cases. OBJECTIVE: In a cohort of children who were followed from birth until age 6 to 7 years, we examined the relationship between parental onset (ie, childhood and adulthood) and duration of atopic disease (ie, persistent disease) and the risk of pediatric atopic disease. Our hypothesis was that different parental disease patterns would be important to pediatric risk of disease. METHODS: Data from 476 families in the ongoing Childhood Allergy Study in Detroit, Mich, were analyzed by using logistic regression. We examined the association between parental patterns of disease and disease onset in their children. Results Father's disease history, particularly asthma history, was more strongly related to pediatric outcomes than mother's history. Asthma status in the fathers, whether it was childhood-only, adulthood only, or persistent, was associated with current asthma in the children. Childhood-only and persistent asthma in fathers conferred a higher risk of atopy in the study children, whereas adulthood-only disease did not. There was also a significant relationship between persistent allergy in the father and atopy in the study children. CONCLUSION: Our data support the hypothesis that there are complex inheritance patterns for allergy and asthma. Therefore, a detailed family history of atopy, including childhood and adulthood experiences, is critical to identifying and classifying risk and disease phenotypes. PMID- 15536409 TI - Can bacterial endotoxin exposure reverse atopy and atopic disease? AB - Studies have shown that endotoxin exposure in childhood is associated with a reduced risk of atopy and atopic asthma. It is commonly assumed that these effects only occur in early life. However, recent epidemiologic studies suggest that immune deviation might take place throughout life. Assuming that the immune system is not fixed after the first years of life, we hypothesize that endotoxin exposure might not only inhibit the development of atopic sensitization and disease at any time throughout life but might also reverse this process. This novel extension of the hygiene hypothesis is primarily based on the indirect evidence of several epidemiologic observations showing a reduction in atopy in adults highly exposed to endotoxin that is unlikely to be explained by protective effects alone. In addition, some animal studies demonstrated the potential of endotoxin to downregulate pre-existing airway eosinophilia and hyperreactivity. However, there is currently little direct evidence that endotoxin might reverse atopy and allergic diseases. Observational studies and randomized trials to test this hypothesis could ultimately lead to the development of novel treatments for atopic diseases, such as allergic asthma, hay fever, and eczema. PMID- 15536410 TI - Mitogen-activated protein kinase pathways: therapeutic targets in steroid resistance? PMID- 15536411 TI - Superantigen-induced corticosteroid resistance of human T cells occurs through activation of the mitogen-activated protein kinase kinase/extracellular signal regulated kinase (MEK-ERK) pathway. AB - BACKGROUND: Microbial superantigens induce human T-cell resistance to corticosteroids. OBJECTIVE: Understanding the molecular pathways resulting in corticosteroid-resistant T cells is important because this condition can complicate the treatment of inflammation. METHODS: The response of human PBMCs to steroids was assessed by using proliferation assays after stimulation with superantigens or anti-CD3 in the presence of various kinase inhibitors. Glucocorticoid receptor alpha (GCRalpha) localization was defined on the basis of intracellular staining. Protein phosphorylation was measured by means of Western blotting. RESULTS: In the current study we found that PBMCs stimulated with superantigen, but not anti-CD3, induced corticosteroid-resistant T cells. However, the purified T cells stimulated either with staphylococcal enterotoxin B (SEB) or anti-CD3 are susceptible to corticosteroid inhibition. These results imply that signals on antigen-presenting cells might act in concert with the T cell receptor to cause steroid resistance. Blockade of CD40-CD40 ligand interaction had no effect on superantigen-induced corticosteroid resistance. However, CD28 costimulation with T-cell receptor activation induced corticosteroid resistance of human T cells in a dose-dependent manner. Superantigen stimulation, compared with anti-CD3 stimulation, was found to induce a more rapid and sustained phosphorylation of mitogen-activated extracellular signal-regulated kinase (ERK). Treatment with PD98059 and UO126 (specific mitogen activated protein kinase kinase [MEK]/ERK inhibitors), but not a p38 inhibitor or a c-Jun N-terminal kinase inhibitor, restored the response to steroids, as indicated by proliferation assays. Furthermore, purified ERK1 and ERK2 were able to phosphorylate recombinant human GCRalpha directly in an in vitro kinase assay. Of note, superantigen-induced corticosteroid resistance was associated with abrogation of GCRalpha nuclear translocation. This effect could be reversed by treatment with MEK/ERK pathway inhibitors. CONCLUSIONS: These data are compatible with the hypothesis that superantigen-induced corticosteroid resistance involves the Raf-MEK-ERK1/ERK2 pathway of T-cell receptor signaling, which leads to GCRalpha phosphorylation and inhibition of dexamethasone-induced GCRalpha nuclear translocation. PMID- 15536412 TI - Genetic basis of the latex-fruit syndrome: association with HLA class II alleles in a Spanish population. AB - BACKGROUND: The latex-fruit syndrome is a well-defined disorder whose genetic background has not been elucidated. OBJECTIVE: To study the genetic basis of the latex-fruit syndrome. METHODS: In a case-control study, we have investigated a carefully selected group of patients allergic to latex, searching for association between latex-fruit allergy and HLA class I and II genes, HLA-DR functional groups, and markers IL4-R1 and FcepsilonRI-betaca . RESULTS: Seventy-eight patients allergic to latex without spina bifida, 33% of them also allergic to fruits, were included in our protocol. Skin prick test results with both a commercial latex extract and purified hevein were significantly greater in patients allergic to latex and fruit than in patients allergic to latex and not fruit. A cutoff point of >7 mm for commercial latex skin prick test diagnosed latex-fruit allergy with a sensitivity of 66.7% (95% CI, 41.0-86.6) and a specificity of 83.3% (95% CI, 68.6-93.0) in our series of patients. No significant differences were found regarding HLA class I, IL4-R1 , or FcepsilonRI betaca allele distributions. However, comparison of HLA class II allelic frequencies between patients allergic to latex and fruit and patients allergic to latex and not fruit showed significant associations of latex-fruit allergy with DQB1 *0201 (corrected P value, .001; odds ratio, 7.3; 95% CI, 2.6-20.0), as well as with HLA-DR functional group E (corrected P value, .028; odds ratio, 16.0; 95% CI, 1.9-134.1). When comparing allelic distribution among different subgroups of patients allergic to latex, additional significant associations of latex-fruit allergy with DRB1 *0301 and *0901, and of latex and not fruit allergy with DQB1 *0202, DRB1 *0701 and *1101, were demonstrated. CONCLUSIONS: Latex-fruit allergy is associated with HLA-DQB1 *0201, DRB1 *0301, and *0901, as well as with HLA-DR functional group E, whereas latex-not-fruit allergy is associated with DQB1 *0202, and with both DRB1 *0701 and *1101 alleles. PMID- 15536413 TI - Expression and function of the angiopoietin receptor Tie-2 in human eosinophils. AB - BACKGROUND: Increased vascularity of bronchial mucosa is closely related to the expression of angiogenic factors, which contribute to the pathogenesis of diseases such as asthma bronchiale. OBJECTIVE: Here we examine the effects of the angiogenic growth factors angiopoietin 1 and angiopoietin 2 on eosinophil function in vitro and possible involvement of the angiopoietin receptor Tie-2. METHODS: Eosinophil migration was studied by micropore filter assays. Signaling mechanisms required for angiopoietin-dependent migration were tested by using signaling enzyme blockers. Tie-2 mRNA and receptor expression on the cell surface of eosinophils was demonstrated in RT-PCR and by fluorescence-activated cell sorting analysis. RESULTS: Angiopoietin 1 significantly stimulated eosinophil chemotaxis via activation of phosphodiesterase, phosphatidylinositol 3'-kinase, and tyrosine kinases. The effect on eosinophil migration of angiopoietin 1 was reversed by an antibody against the Tie-2 receptor and by angiopoietin 2. Incubation of eosinophils with angiopoietin 1 abolished the chemotactic effects of vascular endothelial growth factor on human eosinophils via the Tie-2 receptor. Finally, Tie-2 expression by human eosinophils was demonstrated on the transcriptional and protein level. CONCLUSIONS: Data suggest that angiopoietin 1 stimulates directed migration and possibly inhibits vascular endothelial growth factor-induced eosinophil chemotaxis via its Tie-2 receptor, which is expressed by eosinophils. Thus, angiopoietin 1 may play an important role in the modulation of eosinophilic inflammation. PMID- 15536414 TI - Protein kinase Cdelta functions downstream of Ca2+ mobilization in FcepsilonRI signaling to degranulation in mast cells. AB - BACKGROUND: Mobilization of Ca 2+ plays an important role in the degranulation of mast cells. Although events upstream of Ca 2+ mobilization in the regulation of degranulation are relatively well characterized, the downstream mediators of Ca 2+ remain largely unknown. OBJECTIVE: We sought to characterize the downstream signaling mechanism by which Ca 2+ mobilization mediates degranulation in antigen stimulated mast cells. METHODS: The effect of various inhibitors was examined in the antigen-induced or Ca 2+ ionophore A23187-induced degranulation process, and the effect of inhibitors on histamine release was tested in the mouse model of asthma. RESULTS: The delta isoform of protein kinase C (PKC) functions downstream of Ca 2+ in the signaling pathway from FcepsilonRI to degranulation in RBL-2H3 mast cells. Stimulation of cells with either antigen or the Ca 2+ ionophore A23187 induced a rapid translocation of PKC-delta from the cytosol to the cellular membranes, and either treatment with the PKC-delta-specific inhibitor rottlerin or infection with an adenovirus encoding a dominant negative mutant of PKC-delta markedly inhibited degranulation induced with antigen or A23187. Furthermore, both the translocation of PKC-delta and degranulation induced by A23187 were inhibited by prevention of the accumulation of reactive oxygen species normally elicited by the ionophore. Finally, intraperitoneal injection of rottlerin prevented the increase in the concentration of histamine in bronchoalveolar lavage fluid induced by means of antigen challenge in a mouse model of allergic asthma. conclusion: PKC-delta plays an essential downstream mediatory role in the degranulation elicited by Ca 2+ mobilization, and reactive oxygen species mediate the activation of PKC-delta by Ca 2+ in the regulation of degranulation. PMID- 15536415 TI - Cysteinyl leukotriene receptor antagonist regulates vascular permeability by reducing vascular endothelial growth factor expression. AB - BACKGROUND: Inflammation of the asthmatic airway is usually accompanied by increased vascular permeability and plasma exudation. Cysteinyl leukotrienes (cysLTs) potently elicit increased vascular permeability in airways, leading to airway edema. Vascular endothelial growth factor (VEGF) is 1 of the most potent proangiogenic cytokines and also increases vascular permeability so that plasma proteins can leak into the extravascular space. However, the mechanisms by which cysLTs induce increased vascular permeability are not clearly understood. OBJECTIVE: An aim of the current study was to determine the role of the cysLTs, more specifically in the increase of vascular permeability. METHODS: We used a BALB/c mouse model of allergic asthma to examine effects of cysLT receptor antagonists on bronchial inflammation and airway hyperresponsiveness, more specifically on the increase of vascular permeability. RESULTS: These mice develop the following typical pathophysiological features of asthma in the lungs: increased numbers of inflammatory cells of the airways, airway hyperresponsiveness, increased vascular permeability, and increased levels of VEGF. Administration of cysLT receptor antagonists markedly reduced plasma extravasation and VEGF levels in allergen-induced asthmatic lungs. CONCLUSION: These results indicate that cysLT receptor antagonists modulate vascular permeability by reducing VEGF expression and suggest that cysLT receptor may regulate the VEGF expression. PMID- 15536416 TI - A signal transducer and activator of transcription 6 haplotype influences the regulation of serum IgE levels. AB - BACKGROUND: Because of its central role in the IL-4/IL-13 pathway, the intracellular signaling molecule signal transducer and activator of transcription 6 ( STAT6 ) may be crucial for IgE production in asthma and allergy. OBJECTIVE: We analyzed the association between polymorphisms in the STAT6 gene and the regulation of serum IgE levels. Methods In a population of 1120 German schoolchildren (age 9-11 years), we genotyped 6 previously identified polymorphisms spanning the STAT6 gene by using the matrix-assisted laser desorption ionization-time of flight mass spectrometry method. Haplotypes were estimated and population-derived IgE percentiles (50% IgE > 60 IU/mL, 66% IgE > 115 IU/mL, and 90% IgE > 457 IU/mL) were modeled as outcome variables in haplotype-trend regression analysis. RESULTS: Polymorphisms located in intron 2 (C2892T) and the 3' untranslated region (T12888C) significantly and consistently contributed to elevated total serum IgE levels. One STAT6 haplotype showed increased odds ratios of 1.58 (95% CI, 1.08-2.32; P = .020), 1.82 (95% CI, 1.19 2.77; P = .006), and 3.92 (95% CI, 1.93-7.96; P = .0002) for elevated IgE levels at percentiles 50%, 66%, and 90%, respectively. Because C2892T is located within a nuclear factor kappaB transcription factor binding site, a functional role of this polymorphism is very likely. CONCLUSION: The data indicate that within the IL-4/IL-13 pathway, genetic variants in the STAT6 gene significantly contribute to the regulation of serum IgE levels. PMID- 15536417 TI - Sputum and bronchial submucosal IL-13 expression in asthma and eosinophilic bronchitis. AB - BACKGROUND: Nonasthmatic eosinophilic bronchitis is a condition characterized by the presence of eosinophilic airway inflammation in the absence of airflow obstruction or airway hyperresponsiveness. In asthma, the T H 2-type cytokine IL 13 has been implicated in the development of airway inflammation and hyperresponsiveness. Whether the expression of IL-13 is different between these 2 conditions is unknown. OBJECTIVE: We sought to investigate whether IL-13 expression is increased in asthma compared with eosinophilic bronchitis. METHODS: Sputum samples from subjects with mild asthma (n = 30) and eosinophilic bronchitis (n = 15) and normal controls (n = 16) were dialyzed, and IL-13 concentration was measured by ELISA. In a subgroup of these patients, IL-13 protein expression in bronchial biopsies was assessed by immunohistochemistry. RESULTS: The concentration of sputum IL-13 was higher in patients with mild asthma than in normal controls ( P = .03) and in patients with eosinophilic bronchitis ( P = .03). The median (interquartile range) number of IL-13 + cells/mm 2 submucosa was significantly higher in asthma 4 (8) than eosinophilic bronchitis 1.7 (1.9) and normal controls 0.5 (1.1; P = .004). Eighty-three percent of the cells expressing IL-13 in the submucosa were eosinophils, and 8% were mast cells. The median (interquartile range) proportion of eosinophils that expressed IL-13 was higher in the subjects with asthma, 16 (10)%, than those with eosinophilic bronchitis, 7 (3)% ( P = .02). CONCLUSION: The increased expression of IL-13 in asthma compared with eosinophilic bronchitis supports the concept that IL-13 may play a critical role in the pathophysiology of asthma. PMID- 15536418 TI - Similar allergic inflammation in the middle ear and the upper airway: evidence linking otitis media with effusion to the united airways concept. AB - BACKGROUND: Otitis media with effusion (OME) is a chronic inflammatory disease of the middle ear space characterized by the accumulation of fluid. Previous investigations have suggested that the immunopathologic mechanism underlying the development of middle ear effusion in patients with allergy is largely due to the effects of T(H)2 mediators. The composition of the inflammatory substrate in the effusions of allergic otitis media is similar to the late-phase allergic response seen elsewhere in the respiratory tract, such as in asthma and in allergic rhinitis. OBJECTIVE: To determine whether the middle ear compartment may be a component of the united airways in allergic disease by comparing the inflammatory profiles of the middle ear to the upper airway. METHODS: Middle ear effusions, torus tubaris (Eustachian tube mucosa at the nasopharyngeal orifice), and adenoidal tissue biopsies were obtained from 45 patients undergoing simultaneous tympanostomy tube placement for OME and adenoidectomy for adenoid hypertrophy. The cellular and cytokine profiles of each site were investigated by using immunocytochemistry (elastase, CD3, major basic protein) and in situ hybridization (IL-4, IL-5, IFN-gamma mRNA). Atopic status was determined for each patient by using skin prick testing. RESULTS: Eleven of the 45 patients with OME (24%) were atopic. The middle ear effusions of atopic patients had significantly higher levels of eosinophils, T lymphocytes, and IL-4 mRNA + cells ( P < .01) and significantly lower levels of neutrophils and IFN-gamma mRNA + cells ( P < .01) compared with nonatopic patients. The nasopharyngeal tissue biopsies revealed similar cellular and cytokine profiles. CONCLUSION: In atopic patients with OME, the allergic inflammation occurs on both sides of the Eustachian tube, both in the middle ear and in the nasopharynx. The results of this study support the concept that the middle ear may be part of the united airway in atopic individuals. PMID- 15536419 TI - Health effects of air pollution. AB - The general public, especially patients with upper or lower respiratory symptoms, is aware from media reports that adverse respiratory effects can occur from air pollution. It is important for the allergist to have a current knowledge of the potential health effects of air pollution and how they might affect their patients to advise them accordingly. Specifically, the allergist-clinical immunologist should be keenly aware that both gaseous and particulate outdoor pollutants might aggravate or enhance the underlying pathophysiology of both the upper and lower airways. Epidemiologic and laboratory exposure research studies investigating the health effects of outdoor air pollution each have advantages and disadvantages. Epidemiologic studies can show statistical associations between levels of individual or combined air pollutants and outcomes, such as rates of asthma, emergency visits for asthma, or hospital admissions, but cannot prove a causative role. Human exposure studies, animal models, and tissue or cellular studies provide further information on mechanisms of response but also have inherent limitations. The aim of this rostrum is to review the relevant publications that provide the appropriate context for assessing the risks of air pollution relative to other more modifiable environmental factors in patients with allergic airways disease. PMID- 15536420 TI - Characterization of natural Dac g 1 variants: an alternative to recombinant group 1 allergens. AB - BACKGROUND: Production of soluble correctly folded recombinant group 1 allergens has proven to be difficult. Purified natural group 1 allergens could be an alternative for application in immunotherapy. OBJECTIVE: Cloning and expression of recombinant Dac g 1; purification of natural Dac g 1 variants and immunochemical characterization of these molecules. METHODS: Dac g 1 was cloned and expressed in the yeast Pichia pastoris. Hydrophobic interaction (HIC), size exclusion, and/or affinity chromatography were used to purify Dac g 1 from Dactylis glomerata pollen extract. Dac g 1 variants were analyzed by N-terminal sequencing. Immune reactivity was assessed by sandwich ELISA, competitive RIA, RAST (inhibition), and in vitro basophil histamine release tests. RESULTS: Dac g 1 was cloned, revealing up to 98% amino acid sequence homology to other group 1 allergens. Purification of natural Dac g 1 revealed at least 3 variants, with an apparent molecular mass (Mr) on SDS-PAGE of 33 kd (HMr), 30 kd (IMr) and 28 kd (LMr). Extraction of IMr Dac g 1 required 0.9% saline, whereas the other 2 variants were also extractable in water. The N-terminus of HMr and IMr Dac g 1 differs at 2 positions, and LMr Dac g 1 was shown to be N-terminally truncated, lacking the first 30 amino acids. The nonretarded fraction of HIC commonly used in group 1 purification protocols does not contain this LMr molecule. IMr Dac g 1 was poorly recognized in 2 of 3 sandwich ELISAs and competitive RIA but demonstrated similar biological activity compared with HMr Dac g 1. CONCLUSIONS: Natural Dac g 1 variants can be separated by extraction of pollen in the presence or absence of saline followed by HIC and size exclusion chromatography. Thus, purified Dac g 1 is an alternative to recombinant group 1 allergens. PMID- 15536421 TI - Effects of wintertime ambient air pollutants on asthma exacerbations in urban minority children with moderate to severe disease. AB - BACKGROUND: Urban minority children with asthma are at higher risk for severe exacerbations leading to hospitalizations and deaths. Because multiple studies have reported associations between air pollution and asthma worsening, elevated levels of air pollution are cited as a possible trigger for increased asthma morbidity in urban areas. Few studies have prospectively followed panels of urban children with asthma to determine whether air pollution levels are associated with clinically relevant outcomes such as asthma exacerbations. OBJECTIVE: To determine the association between levels of ambient air pollutants and asthma exacerbations in urban poor children with moderate to severe asthma. METHODS: A school-based panel of children with difficult-to-control disease was followed over a period of 3 consecutive winters in Denver, Colo. The panel consisted of predominantly urban African American children with moderate to severe asthma. Levels of Environmental Protection Agency criteria air pollutants were measured on a daily basis with concurrent monitoring of lung function, bronchodilator use, symptoms, and asthma exacerbations. RESULTS: After controlling for time-varying factors such as upper respiratory infections and meteorologic factors, a weak association was found between ambient carbon monoxide levels and bronchodilator use. Ozone levels were associated with daytime symptoms only. No association was observed between daily air pollution concentrations and daily levels of FEV 1 , peak flow, nighttime symptom scores, or asthma exacerbations over the 3-year period. CONCLUSION: Ambient levels of Environmental Protection Agency criteria air pollutants in Denver do not lead to clinically significant asthma worsening in urban children with moderate to severe asthma during winter months when children are primarily indoors. PMID- 15536422 TI - Perception of risk associated with asthma research procedures among adolescents, parents, and pediatricians. AB - BACKGROUND: Little empirical data exist about how adolescents with asthma, their parents, and pediatricians view the risks and benefits associated with asthma clinical research. OBJECTIVE: Two studies examined similarities and differences in the perception of risks and benefits associated with asthma research. METHODS: In study I questionnaires were completed by adolescents with asthma and parents at the end of an audio and written presentation of a hypothetical research vignette. In study II adolescents with asthma, their parents, and pediatricians rated the risks and benefits associated with discreet asthma research procedures. RESULTS: In study I adolescents and parents made distinctions in riskiness among the asthma research procedures ( P < .001 and P < .001, respectively). With the exception of venipuncture, rank orders of risk ratings among procedures were similar for both groups. In study II significant differences in risk and benefit ratings for individual procedures were found among respondent groups, including experimental medication, placebo, and venipuncture. Overall, asthma research procedures were viewed as more beneficial than risky ( P < .001). CONCLUSION: Participants generally viewed asthma research procedures as more beneficial than risky. Overall, the relative risk rankings among all respondents were similar. However, there were between-group difference in ratings of risk associated with venipuncture and experimental medication. Parents and adolescents rated the benefit of placebo significantly higher than did pediatricians. PMID- 15536423 TI - Current approach to the diagnosis and management of adverse reactions to foods. PMID- 15536424 TI - Identification and characterization of linear B-cell epitopes of beta-globulin, a major allergen of sesame seeds. AB - BACKGROUND: The increased consumption of foods containing sesame seeds is paralleled by an increase in reported sesame-induced allergic reactions. OBJECTIVE: This study aimed at identifying and characterizing the linear B-cell epitopes of the 14-kd beta-globulin, the major allergen of sesame seed. METHODS: A peptide containing 71 amino acids (peptide B) was previously identified by us as the IgE-binding region on beta-globulin. To determine the amino acid sequence of the IgE-binding sites on peptide B, we synthesized overlapping peptides 20 and 10 amino acid residues long that span the entire length of peptide B, which were offset from each other by 10 and 2 amino acid residues, respectively. Sera from 20 subjects given diagnoses of allergy to sesame beta-globulin served to identify the epitopes by using the dot-blot test. RESULTS: At least 9 different IgE recognition sites were identified on peptide B. Three of them, numbers 2, 3, and 13 (corresponding to amino acids 46-55, 48-57, and 76-86, respectively, in the beta-globulin sequence), appeared to be immunodominant IgE-binding epitopes. Also, these peptides were best recognized in terms of intensity of response. There was no obvious sequence motif shared by the 9 different IgE-binding epitopes of beta-globulin. However, approximately 60% of the amino acids represented in the epitopes are hydrophobic residues. CONCLUSION: Identification of the IgE-binding epitopes might provide a better understanding of the functional role the allergens play in the disease and might have implications for immunodiagnosis and probably immunotherapy. PMID- 15536425 TI - Development of a questionnaire to measure quality of life in families with a child with food allergy. AB - BACKGROUND: Food allergy is potentially severe, affects approximately 5% of children, and requires numerous measures for food avoidance to maintain health. The effect of this disease on health-related quality of life (HRQL) has been documented by using generic instruments, but no disease-specific instrument is available. OBJECTIVE: To create a validated, food allergy-specific HRQL instrument to measure parental burden associated with having a child with food allergy: the Food Allergy Quality of Life-Parental Burden questionnaire. METHODS: After identification of 74 items affecting families with children with food allergy, 88 families were approached for effect scoring. Final items were generated by score results, elimination of redundancies, and content review. Resulting high-effect areas were queried for validation with a 7-point Likert scale. A final instrument including 17 items and 2 expectation of outcome questions was distributed to 352 families for validation. RESULTS: Areas of effect included family/social activities (restaurant meals, social activities, child care, vacation), school, time for meal preparation, health concerns, and emotional issues. Validation steps showed strong internal validity (Cronbach alpha, 0.95) and good correlation with expectation of outcome questions ( r = 0.412; P < .01) and scores on a generic HRQL instrument, the Children's Health Questionnaire-PF50 ( r = -0.36 to -0.4; P < .01). The instrument showed the ability to discriminate by disease burden: parents whose children had multiple (>2) food allergies were more affected than parents whose children had fewer allergies (scores, 3.1 vs 2.6; P < .001). CONCLUSIONS: The Food Allergy Quality of Life-Parental Burden demonstrates strong internal and cross-sectional validity. Its discriminative ability suggests that it will be a useful tool to measure outcomes in treatment studies of food allergy for children. PMID- 15536426 TI - Risk of oral food challenges. AB - BACKGROUND: Oral food challenges are essential to the diagnosis of food allergy; however, little has been reported regarding the risks of performing food challenges in children with suspected food allergy. OBJECTIVE: To examine the risk and reaction severity of failed oral food challenges. METHODS: A retrospective chart review was performed on children who underwent food challenges to milk, egg, peanut, soy, and/or wheat in a university-based pediatric allergy clinic over a 7-year period. RESULTS: Of the 584 challenges completed, 253 (43%) resulted in an allergic reaction. There were 90 milk, 56 egg, 71 peanut, 21 soy, and 15 wheat failed challenges. Of patients who failed, there were 197 (78%) cutaneous, 108 (43%) gastrointestinal, 66 (26%) oral, 67 (26%) lower respiratory, and 62 (25%) upper respiratory reactions. No patients had cardiovascular symptoms. There was no difference between foods in the severity of failed challenges or the type of treatment required to reverse symptoms. All reactions were reversible with short-acting antihistamines +/- epinephrine, beta-agonists, and/or corticosteroids. No children required hospitalization, and there were no deaths. CONCLUSIONS: There are risks associated with food challenges, and the risks are similar for each of the foods studied. Given the benefits that result from a negative challenge, these risks are reasonable when challenges are performed under the guidance of an experienced practitioner in a properly equipped setting. PMID- 15536427 TI - IgE sensitization profiles toward green and gold kiwifruits differ among patients allergic to kiwifruit from 3 European countries. AB - BACKGROUND: Kiwifruits have become a major elicitor of plant food allergy. Until recently, the only species of kiwifruit grown commercially was the common green fleshed Actinidia deliciosa cv Hayward. In 1999, the yellow-fleshed cultivar Actinidia chinensis cv Hort16A was introduced into the international market. OBJECTIVE: We compared the allergen compositions of green and gold kiwifruits and assessed the sensitization patterns of patients with kiwifruit allergy toward both varieties. METHODS: Sera from 90 patients with kiwifruit allergy from Austria, central Italy, and the Netherlands were tested for IgE binding to green and gold kiwifruit protein extracts and to purified actinidin, the major kiwifruit allergen, by ELISA. In addition, ELISA inhibitions and immunoblots were performed with selected sera. Relevant allergens were identified by N-terminal sequencing and immunoblotting with allergen-specific antibodies. RESULTS: IgE immunoblotting showed marked differences in the allergen compositions of green and gold kiwifruit extracts. Phytocystatin, a novel plant food allergen, and a thaumatin-like protein were identified as allergens common for both cultivars. Two allergens with homologies to chitinases were found in gold kiwifruits, whereas actinidin was detected exclusively in green kiwifruits. Patients from Central Europe and central Italy showed distinct sensitization profiles toward green and gold kiwifruit extracts as well as actinidin. Whereas sera from Austrian and Dutch patients mainly recognized green kiwifruit extract and actinidin, almost all Italian sera showed IgE binding to both kiwifruit species, but only half of them contained actinidin-specific IgE. Green and gold kiwifruit extracts were shown to be highly cross-reactive as determined by IgE ELISA inhibition. CONCLUSION: The presence of common allergens and the IgE cross reactivity to green kiwifruit qualifies gold kiwifruit as a potential new allergen source for patients allergic to green kiwifruits. PMID- 15536428 TI - H1 histamine receptor mediates inflammatory responses in human keratinocytes. AB - BACKGROUND: Keratinocytes participate in initiation and amplification of T-cell mediated skin diseases. During these disorders, histamine can be released from both residents skin cells and immigrating leukocytes, and can affect the functions of dendritic cells, monocytes, and lymphocytes. Little information is available on the effects of histamine on keratinocytes. OBJECTIVE: To examine the presence of functional H1 receptors on human keratinocytes and the capacity of histamine to modulate the expression of inflammatory molecules in these cells. METHODS: Primary cultures of resting and cytokine-activated keratinocytes from healthy subjects were analyzed for histamine H1 receptor expression and the production of inflammatory mediators after exposure to histamine receptor agonists and antagonists. RESULTS: Cultured keratinocytes constitutively expressed the H1 receptor mRNA and protein, which was not influenced by IFN gamma, TNF-alpha, or IL-4. H1 but not H2 agonists induced calcium fluxes in keratinocytes. Treatment of keratinocytes with histamine (10 -7 to 10 -4 mol/L) or beta-histine increased the IFN-gamma-induced expression of membrane intercellular adhesion molecule 1 and MHC class I but not MHC class II molecules. Moreover, H1 stimulation promoted basal CC chemokine ligand (CCL)-5/RANTES and GM CSF secretion and augmented IFN-gamma-induced release of the chemokines CCL2/monocyte chemoattractant protein 1, CCL5/RANTES, CCL20/macrophage inflammatory protein 3alpha, and CXC chemokine ligand 10/IFN-induced protein of 10 kd, as well as GM-CSF. Administration of the H1 antihistamine levocetirizine, but not of the H2 antihistamine cimetidine, abolished histamine-dependent expression of all of the investigated proinflammatory molecules in a dose dependent manner (0.01-10 mumol/L). Levocetirizine at higher doses also reduced intercellular adhesion molecule 1, CCL5/RANTES, and GM-CSF release induced solely by IFN-gamma. CONCLUSION: Histamine exerts proinflammatory effects on keratinocytes via the H1 receptor, and these effects are efficiently inhibited by levocetirizine. PMID- 15536429 TI - Onset of action of pimecrolimus cream 1% in the treatment of atopic eczema in infants. AB - BACKGROUND: Data on the efficacy of pimecrolimus cream 1% within the first days of treatment are scarce, as in previous studies, the first postbaseline assessment was performed only after 1 week. OBJECTIVE: We sought to investigate the onset of action of pimecrolimus cream 1% in infants with mild to very severe atopic eczema. METHODS: We used pimecrolimus cream 1% (n = 129) or vehicle cream (n = 66) administered in a double-blind manner for 4 weeks and then open-label pimecrolimus cream 1% for 12 weeks, with a 4-week follow-up period. RESULTS: Pimecrolimus cream 1% reduced the mean Eczema Area and Severity Index at 4 weeks by 71.5% compared with an increase of 19.4% with vehicle ( P < .001). The reduction in the Eczema Area and Severity Index with pimecrolimus cream 1% was significant at day 4 (38.5% vs 17.6% increase with vehicle). Significant improvements in caregivers' assessments of pruritus and sleep loss were observed with pimecrolimus cream 1% by day 2 ( P < .03) and day 3 ( P = .002), respectively, compared with vehicle. Responses to pimecrolimus cream 1% were sustained during the open-label phase, and pimecrolimus cream 1% was well tolerated. Symptoms of atopic eczema returned gradually after discontinuation. CONCLUSION: Pimecrolimus cream 1% was well tolerated and effective in patients with mild to very severe atopic eczema, with rapid onset of action and no disease rebound after discontinuation. PMID- 15536430 TI - Immune responses to mosquito saliva in 14 individuals with acute systemic allergic reactions to mosquito bites. AB - BACKGROUND: Mosquito bite-induced acute systemic allergic reactions are an increasing clinical concern and have not been optimally characterized immunologically. OBJECTIVE: We wanted to study the immunologic basis of these reactions. METHODS: Sera were received from 14 individuals with a history of acute systemic allergic reactions to mosquito bites, defined as the presence of one or more of the following: urticaria, angioedema, wheezing, dyspnea, hypotension, and decrease or loss of consciousness. Ten individuals were from the United States and one each was from Canada, Germany, Japan, and Switzerland. An indirect ELISA was developed to measure specific IgE and IgG antibodies to saliva from 5 common mosquito species with different geographic distributions: Aedes aegypti, Aedes vexans, Aedes albopictus, Anopheles sinensis, and Culex quinquefasciatus. Twenty-nine individuals with negative bite test results from laboratory-reared mosquitoes served as control subjects. RESULTS: Mosquito saliva specific IgE levels to all 5 species were significantly increased in the individuals with systemic allergic reactions compared with the control subjects ( P < .061 for Aedes vexans and P < .008 for the remaining 4 species). By using the mean of the control subjects plus 1 SD as a cut-off level, 11 individuals had positive results to Aedes albopictus and up to 4 additional species; 3 individuals had positive results to only one species. Saliva-specific IgG levels were not significantly increased in the individuals with systemic allergic reactions compared with levels seen in the control subjects ( P > .05). CONCLUSION: Acute systemic allergic reactions to mosquito bites involve mosquito saliva-specific IgE and can be characterized immunologically. Aedes albopictus is the most common species associated with systemic allergic reactions to mosquito bites. PMID- 15536431 TI - Peanut allergy: recurrence and its management. AB - BACKGROUND: Although peanut allergy may recur, the frequency with which this occurs is unknown. OBJECTIVE: The goals of this study were to determine the rate of peanut allergy recurrence, identify risk factors for recurrent peanut allergy, and develop specific recommendations for the treatment of patients with resolved peanut allergy. METHODS: Children who outgrew peanut allergy were evaluated with questionnaires, skin tests, and peanut-specific IgE levels. Patients were invited to undergo a double-blind, placebo-controlled food challenge (DBPCFC) unless the history of a possible recurrence reaction was so convincing that a challenge would be potentially dangerous. RESULTS: Sixty-eight patients were evaluated. Forty-seven patients continued to tolerate peanut, of whom 34 ingested concentrated peanut products at least once per month and 13 ate peanut infrequently or in limited amounts but passed a DBPCFC. The status of 18 patients was indeterminate because they ate peanut infrequently or in limited amounts and declined to have a DBPCFC. After excluding 12 patients originally diagnosed with peanut allergy based solely on a positive skin prick test or peanut-specific IgE level, 3 of 15 patients who consumed peanut infrequently or in limited amounts had recurrences, compared with no recurrences in the 23 patients who ate peanut frequently ( P = .025). The recurrence rate was 7.9 (95% CI, 1.7% to 21.4%). CONCLUSION: Children who outgrow peanut allergy are at risk for recurrence, and this risk is significantly higher for patients who continue largely to avoid peanut after resolution of their allergy. On the basis of these findings, we now recommend that patients eat peanut frequently and carry epinephrine indefinitely until they have demonstrated ongoing peanut tolerance. PMID- 15536432 TI - HLA-DR expression on neonatal monocytes is associated with allergen-specific immune responses. AB - BACKGROUND: The specific mechanisms regulating priming of T-cell immunity to common allergens during early childhood remain to be elucidated, though increasing evidence indicates that antigen-presenting cell function is impaired in childhood. OBJECTIVE: Examine the relationship between HLA-DR expression on monocytes and B cells, allergen-specific T-cell responses at birth, and clinical outcomes at 2 years of age. METHODS: Blood mononuclear cells were obtained from 36 healthy neonates who were followed up clinically to the age of 2 years. Expression of HLA-DR by monocytes and B cells was determined at baseline and after in vitro exposure to IFN-gamma, a cytokine that is known to upregulate the expression of HLA-DR. Mononuclear cells were stimulated with endotoxin or a panel of inhalant and food allergens, and cytokine responses and lymphoproliferation were determined after 1 and 5 days, respectively. RESULTS: The magnitude of HLA DR upregulation on IFN-gamma-stimulated cord blood CD14 + monocytes was consistently correlated with allergen-induced, but not mitogen-induced, lymphoproliferation at birth. HLA-DR upregulation on monocytes was also positively associated with endotoxin-induced IL-12 p70 synthesis (tau = 0.46; P < .001) but inversely related to mite- and ovalbumin-induced IL-13 synthesis ( P = .0006 and P < .003, respectively). HLA-DR expression on unstimulated cord blood monocytes was inversely associated with symptoms of atopic disease at the 2-year follow-up ( P = .015). In contrast, HLA-DR expression on B cells was not associated with these parameters of immune function. CONCLUSIONS: These findings suggest that the maturity of neonatal monocytes and their responsiveness to external stimuli are linked to differing patterns of immune reactivity at birth and to the risk of allergic symptoms in early childhood. PMID- 15536433 TI - Toxic epidermal necrolysis: effector cells are drug-specific cytotoxic T cells. AB - BACKGROUND: Toxic epidermal necrolysis (TEN) is a very rare but extremely severe drug reaction characterized by widespread apoptosis of epidermis with extensive blisters. We previously found drug-specific cytotoxic CD8 T lymphocytes in the blisters of a single patient. OBJECTIVE: To confirm the role of drug specific cytotoxic lymphocytes in a larger series, to test the cytotoxicity on keratinocytes, and to look for cross-reactivity between chemically related drugs. METHODS: The phenotype of lymphocytes present in the blister fluids of 6 patients with TEN was analyzed by flow cytometry. Cytotoxic functions were tested by chromium release assay on a variety of target cells (autologous or MHC class I matched EBV-transformed lymphocytes, autologous keratinocytes) after nonspecific (CD3 monoclonal antibody) or specific (suspected and potentially cross-reactive drugs) activation. RESULTS: Blister lymphocytes were CD8 + HLA-DR + CLA + CD56 + . In all 6 cases, they were cytotoxic after nonspecific activation. A drug specific cytotoxicity was observed in 4 cases (3 related to cotrimoxazole and 1 to carbamazepine) toward lymphocytes. Blister cells also killed IFN-gamma activated autologous keratinocytes in the presence of drug in the 2 patients tested. Blister cells showed a strong immunoreactivity for granzyme B, and cytotoxicity was abolished by EGTA, but not by anti-Fas/CD95, suggesting perforin/granzyme-mediated killing. By using several sulfonamides for testing the specificity of the drug T-cell receptor interaction, we observed cross-reactivity only between 4 structurally closely related medications. CONCLUSION: These results strongly suggest that drug-specific, MHC class I-restricted, perforin/granzyme-mediated cytotoxicity probably has a primary role in TEN. PMID- 15536434 TI - Synergistic effects of fluticasone propionate and salmeterol on in vitro T-cell activation and apoptosis in asthma. AB - BACKGROUND: In asthma T cells are characterized by an increased activation state and by reduced apoptosis. OBJECTIVE: Because the clinical efficacy of inhaled corticosteroids combined with long-acting beta 2 -agonists has been widely demonstrated in asthma, we studied, in vitro, the effect of fluticasone propionate (FP) and salmeterol alone and in combination on the activation and apoptosis of peripheral blood T cells (PBTs), on the expression of phosphorylated nuclear factor kappaB inhibitor (IkappaBalpha), and on the nuclear translocation of glucocorticoid receptor (GR) in PBTs from asthmatic subjects. METHODS: Apoptosis was evaluated on the basis of annexin V binding, whereas the expression of caspases 8 and 3 and phosphorylated IkappaBalpha, as well as the nuclear translocation of the GR, were evaluated by means of Western blot analysis. RESULTS: FP alone increases and salmeterol alone does not affect T-cell apoptosis. The combination of FP and salmeterol significantly increases PBT apoptosis in comparison with FP alone. FP at the lower concentration, when combined with salmeterol, is equivalent to FP at the higher concentration in inducing PBT apoptosis. The synergy in the induction of cell apoptosis is associated with more efficient activation of caspases 8 and 3. FP plus salmeterol is also able to synergistically reduce the expression of phosphorylated IkappaBalpha, thus limiting nuclear factor kappaB activation. The synergy was related to an increased nuclear translocation of the GR. CONCLUSION: This study shows that the combination of FP and salmeterol is able to control PBT activation in asthmatic patients more efficiently than FP alone and with a lower concentration of steroids. PMID- 15536435 TI - Peltier effect-based temperature challenge: an improved method for diagnosing cold urticaria. PMID- 15536436 TI - Lack of association of GPRA (G protein-coupled receptor for asthma susceptibility) haplotypes with high serum IgE or asthma in a Korean population. PMID- 15536437 TI - Occupational asthma caused by shark cartilage dust. PMID- 15536438 TI - Oren C. Durham. PMID- 15536439 TI - A history of pollen mapping and surveillance: the relations between natural history and clinical allergy. PMID- 15536440 TI - The National Allergy Bureau: pollen and spore reporting today. PMID- 15536442 TI - The Aerocrine exhaled nitric oxide monitoring system NIOX is cleared by the US Food and Drug Administration for monitoring therapy in asthma. AB - The Aerocrine exhaled nitric oxide (NO) monitoring system NIOX was cleared by the US Food and Drug Administration for clinical application in patients with asthma in May 2003. The fractional concentration of exhaled NO has been extensively researched as a marker of airway inflammation in asthma and other diseases and is now poised to enter clinical application. The American Thoracic and European Respiratory Societies' current guidelines recommend measurement at constant expiratory flow, which is difficult for some adults and children. The NIOX NO monitoring system was designed to facilitate standardized measurement according to guidelines. A clinical study was performed together with in vitro testing to obtain clearance. Exhaled NO levels were measured in unstable steroid-naive adult and pediatric asthmatic subjects and again after a 2-week treatment with inhaled corticosteroids. Exhaled NO levels decreased highly significantly, with 95% confidence limits for the decrease of -40% to -60% accompanied by clinical improvement. This trial, together with extensive in vitro testing, led to the clearance of NIOX by the US Food and Drug Administration. This article in the journal's "New products" feature section will describe background material regarding exhaled NO, special features of the NIOX NO monitoring system, and how this tool can be incorporated into clinical asthma management. PMID- 15536443 TI - Introduction: Stuffy is also related to Sleepy and Grumpy--the link between rhinitis and sleep-disordered breathing. PMID- 15536444 TI - Inflammatory mediators in allergic rhinitis. AB - Allergic rhinitis (AR) is part of a systemic disease complex. There is a close relationship between AR and asthma, which has led to the "one airway, one disease" concept. Both conditions share common immunopathology and pathophysiology. In patients with AR, allergen-triggered early and late responses are mediated by a series of inflammatory cells. Within minutes of contact with allergen, IgE-sensitized mast cells degranulate, releasing both preformed and newly synthesized mediators. Immunologic processes in both nasal and bronchial tissue involve T H 2 lymphocytes and eosinophils. Eosinophils are the predominant cell in the chronic inflammatory process characteristic of the late-phase allergic response. Eosinophils release an array of proinflammatory mediators, including cysteinyl leukotrienes, cationic proteins, eosinophil peroxidase, and major basic protein, and might serve as a major source of IL-3, IL-5, GM-CSF, and IL-13. Neuropeptides also appear to contribute to the pathophysiology of AR symptoms. Both AR and asthma exhibit marked day-night variation in symptom severity. Acknowledging both the chronobiology of AR and circadian rhythm dependent attributes of antiallergy medications might enhance the beneficial effects of allergy therapies. PMID- 15536445 TI - The correlation between allergic rhinitis and sleep disturbance. AB - Nasal congestion, a common symptom related to allergic rhinitis (AR), often is associated with poor sleep quality, leading to decreased learning ability, decreased productivity at work or school, and a reduced quality of life. The release of inflammatory mediators and activation of inflammatory cells results in nasal congestion, causing disrupted sleep and subsequent daytime somnolence. Therefore it is important to treat AR with medications that improve congestive symptoms without exacerbating sedation. Second-generation antihistamines and anticholinergic drugs are well tolerated but have little effect on congestion and therefore are limited in their ability to reduce AR-associated daytime somnolence. However, intranasal corticosteroids reduce congestion, improve sleep and sleep problems, and reduce daytime sleepiness, fatigue, and inflammation. Recently, montelukast, a leukotriene receptor antagonist, has joined the approved therapies for AR. Montelukast significantly improves both daytime and nighttime symptoms. AR treatment should endeavor to improve daytime and nighttime symptoms, sleep, and productivity, thereby improving quality of life. PMID- 15536446 TI - Pharmacologic approaches to daytime and nighttime symptoms of allergic rhinitis. AB - Allergic rhinitis is associated with sleep disturbances, daytime somnolence, and fatigue. The exact relationship between rhinitis and sleep disturbance is unknown; however, both the symptoms and underlying pathology of allergic rhinitis can interfere with sleep quality. Nasal congestion, which has been shown to cause sleep-disordered breathing, is thought to be primarily responsible for rhinitis related sleep disorders. The severity of nasal congestion follows a circadian rhythm, being worst at night and in the early morning. Chronotherapy is the study of the effects of administration time on the safety and efficacy of drug therapy based on circadian influences on the pharmacokinetics and pharmacodynamics of medications. Chronotherapy studies in allergic rhinitis suggest there are benefits to nighttime dosing of antiallergy medications. For example, the antihistamine mequitazine has shown improved efficacy when administered in the evening compared with morning dosing. More study is needed to determine whether this is a class effect. Leukotriene receptor antagonists are indicated for evening administration; these drugs significantly improve nighttime rhinitis symptoms. Intranasal corticosteroids administered in the morning have demonstrated efficacy in improving nighttime symptoms; however, it is unknown whether evening administration would improve their effects on nocturnal rhinitis symptoms. Because of the significant detrimental effects of nocturnal rhinitis symptoms on quality of life, allergic rhinitis therapies should be evaluated for efficacy in ameliorating nighttime symptoms. PMID- 15536453 TI - Common congenital heart disorders in adults. PMID- 15536454 TI - St John's wort, depression, and catecholamines. PMID- 15536455 TI - Effects of systemic endothelin A receptor antagonism in various vascular beds in men: in vivo interactions of the major blood pressure-regulating systems and associations with the GNB3 C825T polymorphism. AB - OBJECTIVE: We used the orally available endothelin A (ETA) receptor antagonist darusentan to characterize interactions between the major blood pressure regulating systems in healthy men. Mediators of the endothelin system, the sympathetic nervous system, and the renin-angiotensin system act via G protein coupled receptors with a possible involvement of the G-protein beta3 subunit (GNB3) C825T polymorphism. We studied the influence of this polymorphism on the responses to ETA antagonism in the presence of endothelin 1 (ET-1), norepinephrine (NA), and angiotensin II (ANGII). METHODS: Thirty-seven individuals were included in a randomized, double-blind, placebo-controlled, crossover trial with 100 mg darusentan. Systemic hemodynamics and plasma ET-1, NA, and ANGII concentrations were assessed. Local studies were performed in the dorsal hand veins (n=18) and skin microcirculation (n=12), respectively. RESULTS: Darusentan lowered systolic and diastolic blood pressure ( P <.001 versus placebo) without any differences according to genotype (mean maximum Delta systolic blood pressure, -7 +/- 2 mmHg for CT/TT versus -5 +/- 3 mmHg for CC, P=.37; mean maximum Delta diastolic blood pressure, -3 +/- 2 mmHg for CT/TT versus -4 +/- 2 mmHg for CC, P=.96). Venoconstriction to ET-1 and NA was not affected by ET A blockade in either group; however, carriers of the 825T allele demonstrated a markedly enhanced venoconstriction to ET-1 and NA (median effective concentration [ED50] for ET-1 after darusentan [placebo]: 2.5 +/- 0.2 pmol/min for CT/TT [2.7 +/- 0.3 pmol/min], P=.42; 3.9 +/- 0.6 pmol/min for CC [4.6 +/- 0.3 pmol/min], P=.42; P=.046 [P=.0005] for CT/TT versus CC) (ED50 for NA after darusentan [placebo]: 5.2 +/- 1.2 ng/min for CT/TT [7.3 +/- 1.2 ng/min], P=.20; 32.9 +/- 7.1 ng/min for CC [19.7 +/- 5.5 ng/min], P=.75; P=.0008 [P=.026] for CT/TT versus CC). Darusentan dilated veins at baseline in CC homozygous subjects (+0.21 +/- 0.05 mm, P=.004 versus placebo). Systemic ET A antagonism inhibited constriction to ET-1 and also to NA and ANGII in the skin microcirculation without differences according to genotype (ET-1, P=.017 for all individuals versus placebo; NA, P=.0005; and ANGII, P=.002). CONCLUSION: GNB3 C825T allele carrier status did not influence systemic hemodynamic or local vascular responses to ET A blockade with darusentan in young, healthy men. However, it determined venoconstriction to exogenous ET-1 and NA. Darusentan markedly inhibited not only ET-1-induced but also NA-induced and ANGII-induced vasoconstriction in the skin microcirculation. In contrast, it had no effects on either ET-1-mediated or NA-mediated venoconstriction, indicating that, in the presence of high local ET-1 concentrations, constrictive endothelin B receptors may be of greater importance in the venous vasculature than has been recognized so far. PMID- 15536456 TI - Effects of cytochrome P450 2C9 polymorphisms on phenprocoumon anticoagulation status. AB - OBJECTIVE: Our objective was to assess whether there is an association between the presence of allelic variants of the gene for cytochrome P450 (CYP) 2C9 and anticoagulation problems during the initial phase of phenprocoumon treatment. METHODS: A prospective follow-up study was performed at 2 anticoagulation clinics in The Netherlands. Included subjects started phenprocoumon during the study period, had their first check of the international normalized ratio (INR) on the third or fourth day of therapy, and had an indication for the low therapeutic range (INR, 2.0-3.5). CYP2C9 genotypes ( CYP2C9*1 , CYP2C9*2 , and CYP2C9*3 ) were assessed, and data on indication, INR checks, comedication, and comorbidity were collected. RESULTS: After genotyping, 284 subjects were available for analysis. Of these, 186 (65.5%) were homozygous carriers of the CYP2C9 wild-type allele ( CYP2C9*1/*1 ), 61 (21.5%) were carriers of the CYP2C9*2 allele, and 37 (13.0%) were carriers of the CYP2C9*3 allele. Compared with homozygous CYP2C9*1/*1 subjects, carriers of CYP2C9*2 or *3 had an increased risk of severe overanticoagulation (INR >6.0). The hazard ratio for CYP2C9*2 versus CYP2C9*1/*1 was 3.09 (95% confidence interval [CI], 1.56 to 6.13; P=.001), and the hazard ratio for CYP2C9*3 versus CYP2C9*1/*1 was 2.40 (95% CI, 1.03 to 5.57; P=.042). Carriers of CYP2C9*2 also had a lower chance to achieve stability in the follow up period. The hazard ratio for CYP2C9*2 versus CYP2C9*1/*1 was 0.61 (95% CI, 0.43 to 0.85; P=.003). Carriers of the CYP2C9*2 or *3 allele needed a significantly lower phenprocoumon dosage compared with homozygous CYP2C9*1/*1 subjects. CONCLUSION: The presence of at least 1 CYP2C9*2 or *3 allele in phenprocoumon users is associated with an increased risk of severe overanticoagulation. Similar to warfarin and acenocoumarol, phenprocoumon had a lower dosage requirement in carriers of CYP2C9*2 or *3 compared with that in CYP2C9 wild-type subjects. PMID- 15536457 TI - A variant 2677A allele of the MDR1 gene affects fexofenadine disposition. AB - BACKGROUND AND OBJECTIVES: There have been considerable disagreements regarding the influence of MDR1 (ABCB1) polymorphisms on the disposition of P-glycoprotein (P-gp) substrates. We speculated that the unknown function of the A allele of exon 21 G2677T/A (Ala893Ser/Thr) provides one of the reasons for the contradictory results. This study was performed to clarify the effects of major MDR1 gene polymorphisms, including a variant A allele in exon 21, on fexofenadine pharmacokinetics. METHODS: We investigated the occurrence of 3 high-frequency single-nucleotide polymorphisms (SNPs) in exons 12 (C1236T), 21 (G2677T/A), and 26 (C3435T) of the MDR1 gene in 232 healthy Koreans, using a polymerase chain reaction-restriction fragment length polymorphism method, and performed haplotype analysis on these 3 SNPs. A single oral dose of 180 mg fexofenadine hydrochloride was administered to 33 healthy Korean male volunteers, who were divided into 6 groups based on the MDR1 genotype for the G2677T/A polymorphism in exon 21 and the C3435T polymorphism in exon 26. RESULTS: A strong linkage disequilibrium was observed among the 3 SNPs. The frequencies of the 4 major haplotypes, 1236C-2677A 3435C, C-G-C, T-G-C, and T-T-T, were 16.4%, 18.6%, 21.6%, and 32.2%, respectively. Fexofenadine disposition varied considerably among the groups. In the 2677AA/3435CC genotype group (n=3), the values of area under the concentration-time curve from time 0 to 24 hours [AUC(0-24)] were significantly lower (P=.014) than those of the other 5 genotype groups (GG/CC, GT/CT, TT/TT, GA/CC, and TA/CT). As compared with the 2677GG/3435CC subjects, the AUC(0-24) values were 17% lower in the 2677AA/3435CC subjects and 47% higher in the 2677TT/3435TT subjects (GG/CC versus AA/CC versus TT/TT, 4017 +/- 1137 ng . h/mL versus 3315 +/- 958 ng . h/mL versus 5934 +/- 2,064 ng . h/mL; P=.018). By stratification for genotypes at position 3435, homozygous 3435TT subjects were found to have significantly higher AUC(0-24) (P=.024) and maximum plasma concentration (P=.040) values than CC subjects [AUC(0-24), 5934 +/- 2064 ng . h/mL versus 3998 +/- 1241 ng . h/mL; maximum plasma concentration, 958 +/- 408 ng/mL versus 673 +/- 242 ng/mL]. CONCLUSIONS: The plasma concentrations of fexofenadine after a single oral administration were lower in 2677AA/3435CC subjects than in subjects with the other 5 genotype combinations of the SNPs of G2677T/A and C3435T. These findings confirm the importance of analyzing MDR1 haplotypes and provide a plausible explanation for the conflicting results regarding the effect of MDR1 polymorphisms on the disposition of P-gp substrates. PMID- 15536458 TI - In vivo assessment of botanical supplementation on human cytochrome P450 phenotypes: Citrus aurantium, Echinacea purpurea, milk thistle, and saw palmetto. AB - OBJECTIVES: Phytochemical-mediated modulation of cytochrome P450 (CYP) activity may underlie many herb-drug interactions. Single-time point phenotypic metabolic ratios were used to determine whether long-term supplementation of Citrus aurantium , Echinacea purpurea , milk thistle (Silybum marianum), or saw palmetto (Serenoa repens) extracts affected CYP1A2, CYP2D6, CYP2E1, or CYP3A4 activity. METHODS: Twelve healthy volunteers (6 women, 6 men) were randomly assigned to receive C aurantium , E purpurea , milk thistle, or saw palmetto for 28 days. For each subject, a 30-day washout period was interposed between each supplementation phase. Probe drug cocktails of midazolam and caffeine, followed 24 hours later by chlorzoxazone and debrisoquin (INN, debrisoquine), were administered before (baseline) and at the end of supplementation. Presupplementation and postsupplementation phenotypic trait measurements were determined for CYP3A4, CYP1A2, CYP2E1, and CYP2D6 by use of 1-hydroxymidazolam/midazolam serum ratios (1 hour sample), paraxanthine/caffeine serum ratios (6-hour sample), 6 hydroxychlorzoxazone/chlorzoxazone serum ratios (2-hour sample), and debrisoquin urinary recovery ratios (8-hour collection), respectively. The content of purported "active" phytochemicals was determined for each supplement. RESULTS: Comparisons of presupplementation and postsupplementation phenotypic ratios suggested that these particular supplements had no significant effect on CYP1A2, CYP2D6, CYP2E1, or CYP3A4 activity. Phytochemical profiles indicated that C aurantium was devoid of the CYP3A4 inhibitor 6',7'-dihydroxybergamottin. Quantities of fatty acids, flavonolignans, and cichoric acid were consistent with label claims for saw palmetto, milk thistle, and E purpurea , respectively. CONCLUSIONS: Botanical supplements containing C aurantium , milk thistle, or saw palmetto extracts appear to pose a minimal risk for CYP-mediated herb-drug interactions in humans. Although the effects of E purpurea on CYP activity were minor, further study into the interaction potential of this botanical is merited. PMID- 15536459 TI - Population one-compartment pharmacokinetic analysis with missing dosage data. AB - OBJECTIVE: Our objective was to develop a population 1-compartment pharmacokinetic (PK) method of analysis to deal with suspect or missing prior dosage history. METHODS: Population PK data from a 1-compartment model with first order elimination and absorption, described by PK parameters clearance, volume of distribution, and absorption rate constant, are simulated. A PK sample is drawn just before a test dose (Dt), followed by a (varying) number of additional samples over 1 interdose interval (tau). For 60% of the subjects, the true history of the scheduled dose (Ds) preceding Dt differs from that prescribed, whereas doses taken before Ds do not. Two settings are evaluated: considerable accumulation of drug in the body (typical drug half-life t1/2 approximately equal to tau) and very little such accumulation (t1/2 approximately equal to tau/5). Precision and bias of several PK analysis methods--Missing Dose Method (MDM), Missing Dose Mixture Method (MDMM) and Extrapolation-Subtraction Method (ESM), all of which essentially do not use prior dose history--are compared with those of the Prescribed Dose Method (PDM), which assumes nominal dosage, and an Ideal Method (IDM), which uses true (but unknown) pre-test dose history. RESULTS: At t1/2 approximately equal to tau, MDM and MDMM are the most precise methods. The accuracy of ESM and PDM is poor. At t1/2 approximately equal to tau/5, no significant differences, in terms of precision or bias, are observed between methods. Misspecification of the structural or statistical model seems not to influence these results. The results of analysis of a real (caffeine) data set are compatible with the findings from the simulations. CONCLUSION: When a test dose is given and a predose baseline observation is taken as part of an "intensive" PK study during outpatient therapy of a 1-compartment drug, an analysis that assumes that the nominal dose history is correct is not robust to past dosage history misspecification, whereas methods that do not do this are robust and reliable. PMID- 15536460 TI - Intravenous and oral alfentanil as in vivo probes for hepatic and first-pass cytochrome P450 3A activity: noninvasive assessment by use of pupillary miosis. AB - INTRODUCTION: Systemic clearance of intravenous (IV) alfentanil (ALF) is an in vivo probe for hepatic cytochrome P450 (CYP) 3A activity, miosis is a surrogate for plasma ALF concentrations, and IV ALF miosis is a noninvasive probe for hepatic CYP3A. This investigation characterized the bioavailability and first pass metabolism of oral ALF and tested the hypotheses that (1) first-pass ALF clearance reflects first-pass CYP3A activity, (2) miosis after oral ALF will reflect intestinal and hepatic CYP3A activity, and (3) miosis can approximate plasma concentration-based pharmacokinetic measures for IV and oral ALF as a noninvasive in vivo probe for hepatic and first-pass CYP3A activity and drug interactions. Results were compared with those for midazolam (MDZ), an alternative CYP3A probe. METHODS: Ten volunteers were studied by use of a randomized, 9-way, crossover design after administration of rifampin (INN, rifampicin) (hepatic and intestinal CYP3A induction), troleandomycin (TAO) (hepatic and intestinal CYP3A inhibition), grapefruit juice (selective intestine CYP3A inhibition), or nothing (control). For each condition, they received 1 mg IV MDZ and then 15 microg/kg IV ALF, as well as 3 mg oral MDZ and then oral ALF (23 or 60 microg/kg) on another day. Plasma concentrations were determined by liquid chromatography-mass spectrometry. Dark-adapted pupil diameters were measured coincident with blood sampling. ALF effect was analyzed similarly to concentration to yield an effect "clearance" (Dose/Area under the pupil diameter change versus time curve). RESULTS: Bioavailability (Foral), hepatic extraction (EH), and intestinal availability (FG) were 0.26 +/- 0.08, 0.52 +/- 0.09, and 0.56 +/- 0.20, respectively, for MDZ and 0.42 +/- 0.15, 0.28 +/- 0.09, and 0.56 +/- 0.18, respectively, for ALF. Oral clearance (CL/F) was 34.7 +/- 12.8 and 10.9 +/- 3.5 mL.kg -1.min -1 , respectively, for MDZ and ALF. After rifampin, TAO, and grapefruit juice, ALF F oral was 0.04 +/- 0.02 (P <.05, versus control), 0.99 +/- 0.18 (P <.05, versus control), and 0.62 +/- 0.18 (P <.05, versus control), respectively; E H was 0.69 +/- 0.14 (P < .05, versus control), 0.04 +/- 0.01 (P <.05, versus control), and 0.26 +/- 0.08, respectively; F G was 0.16 +/- 0.10 (P <.05, versus control), 1.0 +/- 0.2 (P <.05, versus control), and 0.85 +/- 0.30 (P <.05, versus control), respectively; CL/F was 339 +/- 233 (P <.05, versus control), 0.62 +/- 0.26 (P <.05, versus control), and 6.7 +/- 2.5 (P <.05, versus control), respectively, and effect clearance was 2.1 +/- 1.1 (P <.05, versus control), 0.087 +/- 0.056 (P <.05, versus control), and 0.54 +/- 0.30 (0.73 +/- 0.43 mg.mm -1.h -1 in controls), respectively. There were significant correlations between ALF and MDZ systemic clearances (r2= 0.92), EH (r2=0.93), and CL/F (r2= 0.97), as well as between oral ALF effect (miosis) clearance and oral clearance (r2=0.59). CONCLUSIONS: ALF and MDZ have similar intestinal extraction but low and intermediate hepatic extraction, respectively. Systemic and oral clearances of ALF are excellent in vivo probes for hepatic and first pass CYP3A activities and drug interactions. Miosis was an acceptable surrogate for plasma ALF. ALF miosis may be a suitable noninvasive in vivo probe for both hepatic and first-pass CYP3A. PMID- 15536461 TI - Age and gender effects on the pharmacokinetics and pharmacodynamics of triazolam, a cytochrome P450 3A substrate. AB - Sixty-one healthy men and women, aged 20 to 75 years, received single 0.25-mg doses of triazolam, a cytochrome P450 (CYP) 3A substrate benzodiazepine, and placebo in a double-blind crossover study. Among women, age had no significant effect on area under the triazolam plasma concentration curve (AUC) (Spearman r=0.14, P=.44) or clearance (r =-0.09, P=.62). Among men, AUC increased (r=0.43, P <.02) and clearance declined (r=-0.42, P <.02) with increasing age. Gender differences in triazolam kinetics were not apparent. Compared with placebo, triazolam impaired digit-symbol substitution test performance, increased observer rated sedation, impaired delayed recall of information learned at 1.5 hours after dosing, and increased electroencephalographic beta amplitude. Among men, mean values of relative digit-symbol substitution test decrement (P <.002) and observer-rated sedation (P <.05) were significantly greater in elderly subjects compared with young subjects. Age-dependent differences among women reached significance for observer-rated sedation (P <.02). A combination of higher plasma levels and increased intrinsic sensitivity explained the greater pharmacodynamic effects of triazolam in elderly subjects. Although the findings are consistent with reduced clearance of triazolam in elderly men, individual variability was large and was not explained by identifiable demographic or environmental factors. PMID- 15536462 TI - Influence of St John's wort on catecholamine turnover and cardiovascular regulation in humans. AB - BACKGROUND: St John's wort (Hypericum perforatum) is a popular over-the-counter antidepressant. Its antidepressive effect has been attributed in part to inhibition of monoamine transporters and monoamine oxidase, on the basis of in vitro studies. METHODS: In a double-blind, randomized, placebo-controlled, crossover study, 16 healthy subjects (11 men and 5 women; mean age, 31 +/- 5 years) ingested either St John's wort (300 mg three times daily) or placebo for 7 days. Imipramine treatment (50 mg three times daily) in 7 subjects served as a positive control. After treatment, physiologic and biochemical tests included cardiovascular reflex testing, graded head-up tilt testing, and plasma catecholamine determinations. RESULTS: St John's wort had no effect on blood pressure, heart rate, heart rate variability, or blood pressure variability, regardless of the test condition. St John's wort had no effect on plasma concentrations of norepinephrine and its main metabolite, dihydroxyphenylglycol, whereas plasma dihydroxyphenylacetic acid (DOPAC; the main metabolite of dopamine) concentrations increased in every subject (1661 +/- 924 pg/mL versus 1110 +/- 322 pg/mL with placebo, P=.04). In contrast, imipramine increased resting blood pressure (124 +/- 10 mmHg/71 +/- 5 mmHg versus 110 +/- 8 mmHg/61 +/ 6 mmHg with placebo, P=.005 for systolic values and P=.003 for diastolic values) and heart rate (74 +/- 7 beats/min versus 62 +/- 6 beats/min with placebo, P=.005) and elicited a marked orthostatic tachycardia (increase in heart rate of 43 +/- 17 beats/min versus 26 +/- 8 beats/min with placebo, P=.006). CONCLUSIONS: Our findings challenge the concept that St John's wort elicits a major change in norepinephrine uptake or monoamine oxidase activity in vivo. The consistent increase in plasma DOPAC concentrations might suggest a novel mode of action or an inhibitory effect on dopamine beta-hydroxylase that should be followed up. We propose that a combination of physiologic and biochemical profiling may help better define the mode of action and potential side effects of herbal remedies. PMID- 15536463 TI - A phase I trial of pharmacologic modulation of irinotecan with cyclosporine and phenobarbital. AB - The anticancer agent irinotecan has been demonstrated to improve the survival rate in patients with metastatic colorectal cancer. Its usage has been limited by severe toxicity. To modulate irinotecan pharmacokinetics and reduce the prevalence of severe toxicity, patients were treated with cyclosporine (INN, ciclosporin) and the irinotecan dose was increased from 25 to 72 mg/m2 weekly. Phenobarbital was then added, allowing dose escalation to 144 mg/m2. Dose limiting toxicities were neutropenia and diarrhea. Irinotecan was well tolerated at the recommended phase II dose of 120 mg/m2, with a 6% prevalence of grade 4 neutropenia and an 18% prevalence of grade 3 diarrhea. Cyclosporine increased 7 ethyl-10-hydroxycamptothecin (SN-38) area under the concentration-time curve (AUC) by 23% to 630% and reduced irinotecan clearance by 39% to 64% when compared with historical controls. Phenobarbital increased irinotecan clearance by 27% (P < or =.001) and reduced SN-38 AUC by 75% (P < or =.001) when compared with patients treated with cyclosporine alone. Five partial responses were observed. Pharmacokinetic modulation of irinotecan with cyclosporine and phenobarbital has been demonstrated; further studies are necessary to evaluate whether this strategy improves the therapeutic index. PMID- 15536464 TI - Postmarketing surveillance for drug safety. PMID- 15536465 TI - Inhibition of acetylcholinesterase in patients receiving irinotecan (camptothecin 11). PMID- 15536466 TI - Tumor-based pharmacokinetics has greater significance for anticancer drugs than does blood-based pharmacokinetics. PMID- 15536467 TI - Drug interaction of tizanidine and fluvoxamine. PMID- 15536471 TI - Physical activity questionnaire for adolescents validated against doubly labelled water. AB - OBJECTIVE: To validate a physical activity questionnaire for adolescents (PAQA) adapted from the International Physical Activity Questionnaire (IPAQ). DESIGN: Energy expenditure was measured during a 14-day period with doubly labelled water (DLW). PAQA was administered as an interview at the end of the period, asking for physical activity in school, during transportation and leisure-time, during a habitual week. Energy expenditure (EE(PAQA)) was calculated as the product of total physical activity + sleep and predicted resting metabolic rate, and was compared to energy expenditure from DLW (EE(DLW)), thermic effect of feeding excluded. SETTING: Participants were recruited from grade 9 in a compulsory school in Goteborg, Sweden. All data were collected at school, and distribution of DLW and measuring of resting metabolic rate were performed at Sahlgrenska University Hospital. SUBJECTS: A total of 33 adolescents (16 girls, 17 boys) 15.7 (0.4) y performed all measurements. RESULTS: : For the whole group, PAQA underestimated energy expenditure by 3.8 (1.7) MJ (P<0.001). There was a strong correlation (r = 0.62, P<0.001) between EE(PAQA) and EE(DLW), but not for boys (r = 0.42, P = 0.090) and girls (r = 0.33, P = 0.22) separately. CONCLUSIONS: PAQA is not able to predict energy expenditure in Swedish adolescents, largely explained by the amount of unreported time. The ability to rank adolescents energy expenditure is questioned because of the gender effect, although we found a strong correlation for the whole group. PMID- 15536472 TI - Effects of pinitol isolated from soybeans on glycaemic control and cardiovascular risk factors in Korean patients with type II diabetes mellitus: a randomized controlled study. AB - OBJECTIVE: To assess the effects of soybean-derived pinitol on glycaemic control and cardiovascular risk factors in Korean patients with type II diabetes mellitus. DESIGN: Randomized, double-blind, placebo-controlled, parallel-group trial. SETTING: Pusan Paik Hospital, Pusan, Republic of Korea. INTERVENTIONS: A total of 30 patients with type II diabetes received an oral dose of 600 mg soybean-derived pinitol or placebo twice daily for 13 weeks. RESULTS: Pinitol significantly decreased mean fasting plasma glucose, insulin, fructosamine, HbA1c, and the homeostatic model assessment insulin resistance index (HOMA-IR, P<0.001). Pinitol significantly decreased total cholesterol, LDL-cholesterol, the LDL/HDL-cholesterol ratio, and systolic and diastolic blood pressure and increased HDL-cholesterol (P<0.05). CONCLUSIONS: These data suggest that soybean derived pinitol may be beneficial in reducing cardiovascular risk in Korean type II diabetes. PMID- 15536473 TI - Whole-grain consumption and the metabolic syndrome: a favorable association in Tehranian adults. AB - BACKGROUND: Although dietary guidelines recommend increased intake of grain products to prevent chronic diseases, epidemiologic data regarding whole-grain intake association with metabolic syndrome are sparse. OBJECTIVE: To evaluate the relationship between whole-grain intakes, metabolic syndrome and metabolic risk factors in Tehranian adults. DESIGN: Population-based cross-sectional study. SETTING: Tehran, the capital of Iran. SUBJECTS: A representative sample of 827 subjects (357 men and 470 women) aged 18-74 y. METHODS: Usual dietary intake was assessed using a food frequency questionnaire. The procedure developed by Jacobs et al was used to classify grain products into whole and refined grains. Weight and height were measured according to standard protocols and body mass index was calculated. Fasting blood samples were taken for biochemical measurements and blood pressure was assessed according to standard methods. Hypertriglyceridemia, hypercholestrolemia, high LDL, low HDL and metabolic syndrome were defined according to ATP III guidelines and hypertension based on JNC VI. Diabetes was defined as fasting plasma glucose level of > or = 126 mg/dl or 2-h postchallenge blood glucose level of > or = 200 mg/dl. Subjects were categorized based on quartile cut-points of whole- and refined-grain intake. RESULTS: Mean (+/-s.d.) consumptions of whole and refined grains were 93+/-29 and 201+/-57 g/day, respectively. Both men and women reported higher intakes of refined grain than of whole grains. Compared with subjects in the lower quartile category, those in the upper category of whole-grain intake had lower prevalence of metabolic risk factors. Conversely, those in the higher category of refined-grain intake had higher prevalence of metabolic risk factors, except for diabetes. After controlling for confounders, a significantly decreasing trend was observed for the risk of having hypertriglyceridemia (odds ratios among quartiles: 1.00, 0.89, 0.74, 0.61, respectively), hypertension (1.00, 0.99, 0.93, 0.84) and metabolic syndrome (1.00, 0.84, 0.76, 0.68). Higher consumption of refined grains was associated with higher odds of having hypercholestrolemia (1.00, 1.07, 1.19, 1.23), hypertriglyceridemia (1.00, 1.17, 1.49, 2.01), hypertension (1.00, 1.22, 1.48, 1.69) and metabolic syndrome (1.00, 1.68, 1.92, 2.25). CONCLUSION: Whole grain intake is inversely and refined-grain intake is positively associated with the risk of having metabolic syndrome. Recommendations to increase whole-grain intake may reduce this risk. PMID- 15536474 TI - Pancreatic size in protein energy malnutrition: a predictor of nutritional recovery. AB - BACKGROUND: Pancreatic exocrine dysfunction has been frequently recorded in protein energy malnutrition (PEM) because the pancreas requires optimal nutrition for enzyme synthesis. This pancreatic enzyme insufficiency may play a role in the continuation of PEM. OBJECTIVE: This study was designed to assess the pancreatic head size and exocrine pancreatic functions, namely serum amylase and lipase, in PEM and its subtypes and correlate any defect present with the various clinical and laboratory data of the PEM patients with special emphasis on the effect of nutritional rehabilitation. PATIENTS AND METHODS: A total of 33 cases of PEM; 15 marasmus, 10 kwashiorkor (KWO) and eight marasmic kwashiorkor (MKWO) were recruited from Ain Shams University children's hospital, together with 12 matched controls. The mean age of patients was 11.87 +/- 7.8 months and that of the controls was 14.83 +/- 7.7 months. Detailed history taking and thorough clinical examination with special emphasis on anthropometric measurements were taken for each studied infant as well as laboratory investigations which included; complete blood count, liver and kidney functions and serum amylase and lipase. Ultrasonographic assessment of pancreatic head size was performed for the cases and controls. Nutritional rehabilitation program was carried out for 3-6 months followed by reassessment of the cases. RESULTS: The pancreatic head size values were significantly lower in all subtypes of PEM (1.52 +/- 0.6, 2.73 +/- 0.12 and 3.00 +/- 0.54 cm(3) in the marasmus, KWO and MKWO respectively) compared to the controls (5.13 +/- 2.33 cm(3)). The serum amylase and lipase were also significantly lower in all subgroups of PEM when compared to the controls with significant improvement following nutritional rehabilitation coupled by a significant increase in pancreatic head size too. No significant differences were recorded when we compared the subgroups together except for a significant higher rate of change in serum amylase in edematous patients compared to nonedematous ones. The length of nutritional rehabilitation period, age of the patient, weight and serum albumin were the most determinant factors for pancreatic head size as evident from the multiple regression analysis study. CONCLUSION: The potentially correctable exocrine pancreatic insufficiency in cases of PEM should be carefully thought of when planning the nutritional rehabilitation program for such patients as it could be responsible for the serious continued morbidity issues that they face. We thus recommend that estimation of pancreatic head size and exocrine function should be included in the evaluation of PEM patients and they could also be used as a prognostic parameter. SPONSORSHIP: The patients enrolled were among those admitted and managed freely in the children's Hospital Faculty of Medicine Ain Shams University, including the use of the ultrasound apparatus, while the kits have been purchased by authors who contributed in financing the study. PMID- 15536475 TI - Quantification of allele-specific G-protein beta3 subunit mRNA transcripts in different human cells and tissues by Pyrosequencing. AB - The G-protein 825T allele is associated with altered drug responses while the underlying mechanism is not fully understood. Differential expression of transcripts from the C and T alleles could contribute to this process. The C825T polymorphism located in exon 10 is in close linkage disequilibrium with the A( 350)G promoter single nucleotide polymorphism (SNP) and the C1429T SNP and could therefore serve as a marker for allele-specific expression resulting from the promoter SNP. However, alternative splicing of exon 10 in 825T allele carriers may result in under-represented mRNA transcripts. We, therefore, established a novel method based on the Pyrosequencing technology to quantify allele-specific transcript expression and quantified the allelic variance of the C1429T polymorphism located in the 3'-untranslated region of GNB3. Validation of the method was performed using linear regression analysis of measured versus expected ratios of DNA mixed at different known concentrations as well as determining allele-specific mRNA expression of the partially imprinted IGF-2 gene. We genotyped the C1429T polymorphism of 83 samples derived from six different human tissues and cell lines and quantified mRNA transcripts from different alleles using heterozygous samples. No significantly different transcript amounts from the two alleles were found. There were also no significantly different transcript amounts associated with different G(-350)A genotypes (P>0.05). As a result, we could show that Pyrosequencing provides a sensitive tool to quantify allele specific transcript expression. Our data do not support the hypothesis that differential G-protein activity associated with the C825T SNP results from different transcript amounts associated with specific GNB3 genotypes. PMID- 15536476 TI - A genomewide linkage analysis for prostate cancer susceptibility genes in families from Germany. AB - Prostate cancer is a complex disease with a substantial genetic contribution involved in the disease risk. Several genomewide linkage studies conducted so far have demonstrated a strong heterogeneity of susceptibility. In order to assess candidate regions that are particularly relevant for the German population, we performed a genomewide linkage search on 139 prostate cancer families. A nonparametric method (Zlr scores), using GENEHUNTERPLUS, was applied at 500 markers (panel P1400, deCODE), with an average spacing of 7.25 cM. In the entire family collection, linkage was most evident at 8p22 (Zlr=2.47, P=0.0068), close to the previously identified susceptibility gene MSR1. Further local maxima with Zlr>2 (P<0.025) were observed at 1q, 5q and 15q. In a subgroup of 47 families, which matched the Johns Hopkins criteria of hereditary prostate cancer, suggestive linkage was found on 1p31 (Zlr=3.37, P=0.00038), a previously not described candidate region. The remaining 92 pedigrees, with no strong disease history, revealed a maximum Zlr=3.15 (P=0.00082) at 8q13, possibly indicating a gene with reduced penetrance or recessive inheritance. Our results suggest pronounced locus heterogeneity of prostate cancer susceptibility in Germany. In the present study population, the MSR1 gene could play a significant role. Other conspicuous loci, like 1p31 and 8q13, need further investigation in order to verify their relevance and to identify candidate genes. PMID- 15536477 TI - Polymorphic landscape of the human genome. PMID- 15536478 TI - Evolutionary path to the heart. PMID- 15536479 TI - MVK mutations and associated clinical features in Italian patients affected with autoinflammatory disorders and recurrent fever. AB - Autosomal recessive autoinflammatory disorder caused by mutations of the mevalonate kinase gene (MVK), leading to mild, incomplete MK enzyme deficiency (MKD), has been known so far as Hyper-IgD and periodic fever syndrome (HIDS) and regarded as mostly occurring in Northern Europe. Here we report the results of the molecular characterization of the first Italian series of patients affected with autoinflammatory disorders and periodic fever. A total of 13 different mutations, scattered throughout the MVK coding region, were identified in either homozygous or compound heterozygous state in 15 patients. The mutation leading to the V377I amino-acid change, already described also in other series, resulted the most common with a frequency of 50% of all MKD alleles. Among the other mutations, eight had never been described before, including an interstitial deletion of 19 nucleotides in exon 2. In addition to these nucleotide changes, private and polymorphic MVK variants have been detected in the patients under analysis and checked also in a set of control individuals. Clinical features are reported for each of the 15 MKD patients, and life-threatening infections and systemic amyloidosis presented as unexpected MKD-related complications. Our study demonstrates that MKD is a common cause of recurrent fever also in the Italian population, where it is associated with both a wide spectrum of previously unreported MVK mutations and peculiar phenotypic features. PMID- 15536480 TI - A large-scale study of the random variability of a coding sequence: a study on the CFTR gene. AB - Coding single nucleotide substitutions (cSNSs) have been studied on hundreds of genes using small samples (n(g) approximately 100-150 genes). In the present investigation, a large random European population sample (average n(g) approximately 1500) was studied for a single gene, the CFTR (Cystic Fibrosis Transmembrane conductance Regulator). The nonsynonymous (NS) substitutions exhibited, in accordance with previous reports, a mean probability of being polymorphic (q > 0.005), much lower than that of the synonymous (S) substitutions, but they showed a similar rate of subpolymorphic (q < 0.005) variability. This indicates that, in autosomal genes that may have harmful recessive alleles (nonduplicated genes with important functions), genetic drift overwhelms selection in the subpolymorphic range of variability, making disadvantageous alleles behave as neutral. These results imply that the majority of the subpolymorphic nonsynonymous alleles of these genes are selectively negative or even pathogenic. PMID- 15536481 TI - Darwin's finch beaks, Bmp4, and the developmental origins of novelty. PMID- 15536482 TI - Genetic variation of Ginkgo biloba L. (Ginkgoaceae) based on cpDNA PCR-RFLPs: inference of glacial refugia. AB - Ginkgo biloba, a famous living fossil, is the sole survivor of the genus Ginkgo. To make inferences about the glacial refugia that harbored G. biloba, we examined the genetic structure of eight potential refugial populations and plantations using chloroplast DNA (cpDNA) with eight size variants in the trnK1-trnK2 fragment. The data consist of haplotypes from 158 trees collected from eight localities. The majority of the cpDNA haplotypes are restricted to minor portions of the geographical range. Our results suggest that refugia of G. biloba were located in southwestern China. This area is a current biodiversity hotspot of global importance, and may have been protected from the extremes of climatic fluctuations during the Pleistocene. The Ginkgos on West Tianmu Mountain, which were previously considered to be wild by many researchers, may, instead, have been introduced by Buddhist monks. PMID- 15536483 TI - A quantitative genetics model for viability selection. AB - Viability selection will change gene frequencies of loci controlling fitness. Consequently, the frequencies of marker loci linked to the viability loci will also change. In genetic mapping, the change of marker allelic frequencies is reflected by the departure from Mendelian segregation ratio. The non-Mendelian segregation of markers has been used to map viability loci along the genome. However, current methods have not been able to detect the amount of selection (s) and the degree of dominance (h) simultaneously. We developed a method to detect both s and h using an F2 mating design under the classical fitness model. We also developed a quantitative genetics model for viability selection by proposing a continuous liability controlling the viability of individuals. With the liability model, mapping viability loci has been formulated as mapping quantitative trait loci. As a result, nongenetic systematic environmental effects can be easily incorporated into the model and subsequently separated from the genetic effects of the viability loci. The quantitative genetic model has been verified with a series of Monte Carlo simulation experiments. PMID- 15536484 TI - Mating patterns in a hybrid zone of fire-bellied toads (Bombina): inferences from adult and full-sib genotypes. AB - We present two novel methods to infer mating patterns from genetic data. They differ from existing statistical methods of parentage inference in that they apply to populations that deviate from Hardy-Weinberg and linkage equilibrium, and so are suited for the study of assortative mating in hybrid zones. The core data set consists of genotypes at several loci for a number of full-sib clutches of unknown parentage. Our inference is based throughout on estimates of allelic associations within and across loci, such as heterozygote deficit and pairwise linkage disequilibrium. In the first method, the most likely parents of a given clutch are determined from the genotypic distribution of the associated adult population, given an explicit model of nonrandom mating. This leads to estimates of the strength of assortment. The second approach is based solely on the offspring genotypes and relies on the fact that a linear relation exists between associations among the offspring and those in the population of breeding pairs. We apply both methods to a sample from the hybrid zone between the fire-bellied toads Bombina bombina and B. variegata (Anura: Disco glossidae) in Croatia. Consistently, both approaches provide no evidence for a departure from random mating, despite adequate statistical power. Instead, B. variegata-like individuals among the adults contributed disproportionately to the offspring cohort, consistent with their preference for the type of breeding habitat in which this study was conducted. PMID- 15536485 TI - Population genetic differentiation in taxa of Lotus (Fabaceae: Loteae) endemic to the Gran Canarian pine forest. AB - A survey of allozyme variation at 17 loci in 14 populations representing four taxonomically problematic Gran Canarian pine forest endemic taxa of Lotus (L. genistoides, L. holosericeus, L. spartioides and some taxonomically uncertain populations collected under the designation Lotus sp.) was conducted to examine their diversification and systematic relationships. All groups exhibited high values of genetic variation, although inbreeding was common within populations. Considerable among-population genetic homogeneity was detected, as inferred from low values of Gst within each of the groups. The high population sizes of these taxa and a lack of evidence for isolation by distance or genetic bottlenecks indicate that diversity has accumulated over a long period of environmental stability. The association of high genetic distances with low linear distances, and the substantial increase in the values of Gst when the taxa considered were merged in different combinations hint at an incipient (yet probably taxonomically insufficient) reproductive isolation. The genetic similarity between L. genistoides, L. holosericeus and L. spartioides, together with the different behaviour of the populations collected under the designation Lotus sp., may have important implications for the restructuring of the taxonomy of this group when the ongoing morphological studies are completed. PMID- 15536486 TI - Inheritance of apomeiosis (diplospory) in fleabanes (Erigeron, Asteraceae). AB - Unreduced egg formation (apomeiosis) in flowering plants is rare except when it is coupled with parthenogenesis to yield gametophytic apomixis via apospory or diplospory. Results from genetic mapping studies in diverse apomictic taxa suggest that apomeiosis and parthenogenesis are genetically linked, a finding that is compatible with the conventional rationale that apomeiosis is unlikely to evolve independently because of deleterious fitness consequences. An Erigeron annuus (apomictic) x E. strigosus (sexual) genetic mapping population, however, included a high proportion of plants that were highly apomeiotic (diplosporous) but nonapomictic; that is, they lacked autonomous seed production. To evaluate the function and inheritance of diplospory in Erigeron, a diplosporous triploid (2n=3x=27) seed parent was crossed with a sexual diploid (2n=2x=18) E. strigosus pollen parent to produce an F1 of 31 plants. Chromosome numbers and molecular markers (AFLPs) document the inheritance of the maternal genome through unreduced eggs resulting in recombinant but predominantly (77%) tetraploid F1s (2n=4x=36; 2n+n, B(III)). Quantitative evaluation shows continuous variation in the proportion of diplosporous (vs meiotic) ovules (41-89%) in tetraploid F1s despite the presumed equal genetic contribution from the diplosporous mother. These findings demonstrate the functional independence of diplospory and suggest that variation in the trait in F1s is likely due to segregating paternal modifiers. In addition, of six aneuploid (4x-1, 4x-2) F1s, three lack a subset of maternal AFLP markers. These plants likely arose from aberrant megagametogenesis resulting in the loss of maternal chromatin prior to fertilization. PMID- 15536487 TI - A new approach to the study of genetic variability of complex characters. AB - A new approach to multivariate genetic analysis of complex organismal traits is developed. It is based on examination of the distribution of parental strains and the F1 and F2 hybrids in a multidimensional space, and the determination of the directions corresponding to heterozygosity, epistatic and additive gene effects. The effect of heterozygosity includes variability produced by interaction between and within heterozygous loci. The additive gene effects and the remaining epistatic interactions between the homozygous loci can be visualized separately from the effects of heterozygosity by an appropriate projection of the points in multidimensional space. In all, 20 morphological, physiological and behavioural characters and 21 craniometric measures were studied in crosses between two laboratory rat strains. Linear combinations of craniometric and of morphophysiological characters with a high narrow-sense heritability could be identified. These combinations characterized the organismal stress response, which had been selected for in one of the strains. The prospects for the practical application of the new approach and also for the evaluation of the contribution of the genetic diversity to phenotypic variability in animals in natural populations are discussed. PMID- 15536488 TI - Determining the frequency of open windows in residences: a pilot study in Durham, North Carolina during varying temperature conditions. AB - Air pollution exposures in the residential microenvironment can be significantly affected by air exchange rate (AER). A number of studies have shown that AER in residences is significantly affected by the number and location of open windows and doors. A pilot study was conducted in Durham, North Carolina, to determine whether useful data on open windows and doors could be acquired through a visual survey. The study consisted of 72 2-h survey sessions conducted between October 24, 2001 and March 13, 2003. During the first hour of each session, a technician selected a set of corner residences in one of 48 census tracts and completed a survey form and meteorological measurements for each residence. During the second hour, the technician revisited each residence surveyed during the first hour. The resulting database included data on 2200 "residential visits" (1100 residences times two visits per residence). The technician observed one or more open windows during 20.0 percent of the residential visits. One or more open doors were observed during 13.4 percent of the residential visits; 28.2 percent of the residential visits were associated with at least one open window or door. A series of stepwise linear regression analyses were performed on the data to identify factors associated with open windows and doors. Results of these analyses indicated that the likelihood of one or more windows being opened tended to increase under the following conditions: occupancy at time of visit; session during April, May, or June; high population or housing density; window air conditioning (AC) units; absence of AC; large number of doors; and wind speed above 2 mph. The likelihood of open doors tended to increase under the following conditions: occupancy at time of visit; residence within city limits; session during April, May, or June; detached one-story residence; large number of doors; high housing density; school out; and residence within 10 m of road. Transition probabilities (closed to open and open to closed) were determined for windows and doors by time of day. PMID- 15536489 TI - Association of criteria pollutants with plasma hemostatic/inflammatory markers: a population-based study. AB - To elucidate the health effects of air pollution, the short-term association of criteria pollutants (particles <10 microm in diameter [PM(10)], O(3), CO, NO(2), and SO(2)) with hemostatic and inflammatory markers were examined using a population-based sample of 10,208 middle-age males and females of the biracial cohort of Atherosclerosis Risk in Communities (ARIC) study. For each participant, we calculated the following pollutant exposures 1-3 days prior to the randomly allocated cohort examination date: PM(10), CO, NO(2), and SO(2) as 24-h averages, and O(3) as an 8-h average of the hourly measures. The hemostatic/inflammatory factors included fibrinogen, factor VIII-C, von Willebrand factor (vWF), albumin, and white blood cell count (WBC). Linear regression models were used to adjust for cardiovascular disease (CVD) risk factors, demographic and socioeconomic variables, and relevant meteorological variables. One standard deviation (SD) increment of PM(10) (12.8 microg/m(3)) was significantly (P < 0.05) associated with 3.93% higher of vWF among diabetics and 0.006 g/dl lower of serum albumin among persons with a history of CVD. One SD increment of CO (0.60 p.p.m.) was significantly (P < 0.01) associated with 0.018 g/dl lower of serum albumin. Significant curvilinear associations, indicative of threshold effects, for PM(10) with factor VIII-C, O(3) with fibrinogen and vWF, and SO(2) with factor VIII-C, WBC, and serum albumin were found. This population-based study suggest that the hemostasis/inmflammation markers analyzed, which are linked to higher risk of CHD, are associated adversely with environmentally relevant ambient pollutants, with the strongest associations in the upper range of the pollutant distributions, and in persons with a positive history of diabetes and CHD. PMID- 15536490 TI - Microarray analysis of differentially expressed genes in rat frontal cortex under chronic risperidone treatment. AB - Long-term administration of antipsychotic drugs can induce differential expression of a variety of genes in the brain, which may underscore the molecular mechanism of the clinical efficacy and/or side effects of antipsychotic drugs. We used cDNA microarray analysis to screen differentially expressed genes in rat frontal cortex under 4 weeks' treatment of risperidone (1 mg/kg). Using real-time quantitative PCR, we were able to verify eight genes, whose expression were significantly upregulated in rat frontal cortex under chronic risperidone treatment when compared with control animals. These genes include receptor for activated protein kinase C, amida, cathepsin D, calpain 2, calcium-independent receptor for alpha-latrotoxin, monoamine oxidase B, polyubiquitin, and kinesin light chain. In view of the physiological function of these genes, the results of our study suggest that chronic risperidone treatment may affect the neurotransmission, synaptic plasticity, and proteolysis of brain cells. This study also demonstrates that cDNA microarray analysis is useful for uncovering genes that are regulated by chronic antipsychotic drugs treatment, which may help bring new insight into the molecular mechanism of antipsychotic drugs. PMID- 15536492 TI - Medical comorbidity in a bipolar outpatient clinical population. AB - The presence of medical illnesses among inpatients with bipolar disorder is known to complicate treatment and lengthen hospital stay. However, except for a few specific diseases, little is known about prevalence of medical illnesses in bipolar outpatients and the effect it may have on treatment. The authors sought to assess the presence of medical illnesses in a large outpatient clinical sample of bipolar patients, and the effect that medical illnesses may have on the clinical assessment and treatment of the underlying bipolar disorder. Using the Duke University Medical Center clinical database, the authors categorized the medical diagnoses of 1379 patients who were treated with bipolar disorder from 2001 to 2002 through outpatient psychiatric clinics. The prevalence of medical comorbidities was examined, as well as the effect their presence had on the clinician's assessment of disease severity and time to improvement. As expected, medical comorbidities increased with age. The most common systemic illnesses in bipolar outpatients were Endocrine and Metabolic Diseases (13.6% of the sample), Diseases of the Circulatory System (13.0%), and Diseases of the Nervous System and Sense Organs (10.7%). Significant specific diseases included cardiovascular diseases/hypertension (10.7%), COPD/asthma (6.1%), diabetes (4.3%), HIV infection (2.8%), and hepatitis C infection (1.9%). Clinicians assessed greater severity of illness in patients with increasing numbers of comorbid conditions; however, the time to recovery was not significantly effected by the presence of medical comorbidity. In conclusion, comorbid medical illnesses are common in bipolar outpatients, increasing with age. HIV rates may be increased relative to population norms. Their presence compounds the severity of the illness at time of presentation. PMID- 15536491 TI - Serotonin modulates the suppressive effects of corticosterone on proliferating progenitor cells in the dentate gyrus of the hippocampus in the adult rat. AB - This series of experiments explores the interaction between corticosterone and serotonin (5-HT) in the regulation of cell proliferation in the dentate gyrus of the adult rat. Intracerebroventricular 5,7-DHT (5,7-dihydroxytryptamine) (either 200 or 300 microg) resulted in highly significant depletion of 5-HT as measured by high performance liquid chromatography in the frontal cortex but had no effect on the number of proliferating cells in the dentate gyrus by measuring 5-bromo-2' deoxyuridine (BrdU) and Ki-67 cytochemistry. Treatment with PCPA (p chlorophenylalanine: a tryptophan hydroxylase inhibitor: 300 mg/kg initially followed by 100 mg/kg/day) resulted in reduced proliferation as measured by Ki-67 after 3 days treatment, but not by BrdU uptake, and not after 14 days treatment by either method. In addition, injection of corticosterone (10-40 mg/kg/day) for 8 days significantly reduced proliferation in the dentate gyrus, as expected, measured by both BrdU uptake and Ki-67 immunostaining. Adrenalectomized (ADX) rats with a replacement subcutaneous pellet of corticosterone showed reduced proliferation when given additional corticosterone (10 mg/kg/day for 8 days), but this was prevented by 5-HT depletion (i.c.v. 5,7-DHT). Finally, a dose-response study showed that progressive doses of corticosterone (0-40 mg/kg/day) in ADX rats resulted in diminished suppression of proliferation in 5-HT-depleted compared with 5-HT-intact rats. These results strongly suggest that 5-HT regulates the sensitivity of proliferating cells in the dentate gyrus to corticosterone. PMID- 15536493 TI - Release of oxytocin in the rat central amygdala modulates stress-coping behavior and the release of excitatory amino acids. AB - Previous experiments have indicated that the release of oxytocin (OXT) occurs in various hypothalamic and extrahypothalamic brain areas. In the present study, we investigated in male rats whether swim stress triggers the release of OXT in the central amygdala (CeA), a key area in processing emotions and stress responses. Further, we examined the physiological significance of OXT released within the CeA for behavioral responses during forced swimming as well as effects on the local release of selected amino acids including glutamate, aspartate, arginine, taurine, and GABA, which are thought to modulate processing of emotions. Exposure to a 10-min forced swimming session caused a significant increase in OXT release (200%, p<0.01) within, but not outside, the CeA as monitored by microdialysis. Administration of the OXT receptor antagonist des-Gly-NH2d(CH2)5(Tyr(Me)2Thr4)OVT via inverse microdialysis into the amygdala before and during exposure to swimming reduced the floating time by 55% (p<0.05) and increased the swimming time by 29% (p<0.05) indicative of a more active stress-coping strategy. Simultaneously, local administration of the OXT receptor antagonist caused a significant increase in the stress-induced release of the excitatory amino acids glutamate and aspartate, whereas the basal release of these amino acids remained unchanged. Taken together, these findings demonstrate a significant activation of the oxytocinergic system in the CeA in response to swim stress. Furthermore, our data indicate that OXT receptor-mediated mechanisms within the amygdala are involved in the generation of passive stress-coping strategies, which might be mediated at least in part via its inhibitory influence on the local release of excitatory amino acids during stress. PMID- 15536494 TI - Inhibition of Jun N-terminal kinase (JNK) enhances glucocorticoid receptor mediated function in mouse hippocampal HT22 cells. AB - Mitogen-activated protein kinases (MAPKs), including Jun N-terminal kinase (JNK), promote inflammatory and proliferative responses to infection and other environmental stimuli including stress. Relevant to negative regulation of inflammatory pathways by glucocorticoids and the development of glucocorticoid resistance (observed in inflammatory disorders as well as certain neuropsychiatric disorders such as major depression), activation of JNK has been reported to inhibit glucocorticoid receptor (GR) function. In this study, the role of JNK pathways in modulating GR function was further investigated. Treatment of mouse hippocampal (HT22) cells with the selective JNK inhibitor, SP 600125 (0.1-10 microM), resulted in dose-dependent induction of GR-mediated MMTV luciferase activity. SP-600125 also significantly enhanced dexamethasone-induced MMTV-luciferase activity, while increasing GR binding to the glucocorticoid responsive element, both in the presence and absence of Dex. Similar effects were observed in mouse fibroblast cells (LMCAT), and in HT22 cells treated with a JNK specific antisense oligonucleotide. The induction of GR-mediated function by SP 600125 was not due to altered cytosolic GR binding or GR protein expression or enhancement of GR nuclear translocation as determined by Western blot. Taken together, the data indicate that constitutive expression of JNK plays a tonic inhibitory role in GR function, which is consistent with findings that activation of JNK pathways inhibits GR. The data also identify potential pathways involved in the pathogenesis of the glucocorticoid resistance found in certain chronic immune/inflammatory diseases and subgroups of patients with major depression. Moreover, JNK pathways may represent a therapeutic target for normalization of GR function in these disorders. PMID- 15536495 TI - Single and repeated stress-induced modulation of phospholipase C catalytic activity and expression: role in LH behavior. AB - PI-PLC, a critical enzyme of the phosphoinositide (PI) signaling pathway, mediates many physiological functions in the brain, including cellular plasticity. Stress-induced learned helplessness (LH) in animals serves as a model of behavioral depression. Recently, we observed that repeated stress prolongs the duration of LH behavior in rats, enabling us to compare neurobiologic abnormalities in acute and chronic depression. Here we examine whether LH behavior is associated with alterations in phospholipase C (PLC), and whether repetition of inescapable shock has similar or dissimilar effects on PLC to those of the single-stress paradigm. Rats were exposed to inescapable shock either once on day 1, or twice, on days 1 and 7. Rats were tested for escape latency on days 2 and 4 after day 1 inescapable shock or on days 2, 8, and 14 after day 1 and 7 inescapable shock. PI-PLC activity and mRNA and protein expression of three different PLC isozymes were determined in the frontal cortex and hippocampus. Higher escape latencies were observed in LH rats tested on day 2 after single inescapable shock and on day 14 after repeated inescapable shock. Single inescapable shock reduced PI-PLC activity in the frontal cortex and hippocampus of LH rats. On the other hand, repeated inescapable shock not only reduced PI-PLC activity in these brain areas of LH rats but also selectively decreased the expression of PLC beta1 and PLC gamma1 isozymes. Our results suggest different responsiveness at the level of PI-PLC after single vs repeated stress, and that reductions in PLC may be critical in the pathophysiology of depression and other stress-related disorders. PMID- 15536496 TI - Enhanced Cdk5 activity and p35 translocation in the ventral striatum of acute and chronic methamphetamine-treated rats. AB - The cyclin-dependent kinase Cdk5 and DARPP-32 (dopamine- and cAMP-regulated phosphoprotein of Mr 32 kDa)-dependent signaling have been implicated in the regulation of dopaminergic neurotransmission after chronic cocaine treatment. In this study, we examined if Cdk5 signaling participates in the behavioral and biochemical effect of acute and chronic methamphetamine (METH) treatment. We found that Cdk5 activity and the membrane fraction of p35 protein, a Cdk5 activator, in the ventral striatum increased transiently after an injection of 4 mg/kg METH, while intra-accumbens treatment with a Cdk5 inhibitor, roscovitine, prevented the acute METH-induced locomotor activation. The phosphorylation of DARPP-32 at both Thr75 and Thr34 was differentially regulated after acute METH treatment, but the levels of total Cdk5, p35, and DARPP-32 remained the same. To determine if Cdk5 signaling was associated with behavior sensitization to METH, rats that received repetitive injections of METH (4 mg/kg) for 14 consecutive days were analyzed at withdrawal day 7. The results indicate that Cdk5 activity and p35 translocation in the ventral striatum were upregulated in METH-sensitized rats; treatment with roscovitine in the nucleus accumbens effectively suppressed the 1 mg/kg METH-induced behavioral sensitization. Concomitantly, a decrease in the amount of PP-2A and DARPP-32 phosphorylation at Thr34, but an increase in phosphorylation of DARPP-32/Thr75, was observed in the ventral striatum of sensitized rats. The overall results demonstrate that Cdk5/p35 and downstream signaling in the ventral striatum play a critical role in the effects of acute METH treatment as well as the development of behavioral METH sensitization. PMID- 15536497 TI - Human cocaine-seeking behavior and its control by drug-associated stimuli in the laboratory. AB - Second-order schedules of drug self-administration were developed to incorporate the effects of drug-related environmental stimuli into an animal model of drug abuse, making it more similar to human situations. Ironically, little is known about how human subjects behave under these schedules. In this study, human volunteers with a history of cocaine use worked on a second-order schedule in which every 100th lever response produced an auditory-visual brief stimulus (2 s). The first stimulus produced after 1 h was extended to 10 s and paired with an intravenous injection of cocaine (25 mg). Up to three injections were allowed per session. In different phases of the experiment, presentation of the brief stimulus was discontinued and/or saline solution (placebo) was injected instead of cocaine. Injections of cocaine were found to maintain responding even when the brief stimulus was not presented. Placebo injections alone did not maintain responding. In contrast, the brief stimulus maintained high levels of responding under placebo conditions, even though self-reports indicated that subjects could clearly discriminate that they were not receiving cocaine. These results demonstrate that drug-related environmental stimuli can maintain persistent drug seeking during periods of drug unavailability. As this procedure directly measures the effects of stimuli on drug seeking, it may provide a valuable complement to indirect measures, such as self-reports of craving, that are often used with human subjects. The similarity of the response patterns in humans and animals also supports the use of second-order schedules in animals as a valid model of human drug seeking. PMID- 15536498 TI - Results of a technology demonstration project to compare rapid aquatic toxicity screening tests in the analysis of industrial effluents. AB - The results of a 'BioWise' demonstration project to assess the comparative sensitivity and practicality of seven new assays for the direct assessment of ecotoxicity in industrial effluents are presented. In addition the aim of the project was to validate the results of the new assays against benchmark data generated from non-proprietary, rapid, microplate screening assays using the regulatory species; freshwater crustacean Daphnia magna and green algae Selenastrum capricornutum, chosen in view of their environmental relevance. The new commercial test assays were: Daphnia magna, Selenastrum capricornutum and Thamnocephalus platyurus Toxkits supplied by Vickers Laboratories Ltd, containing dormant, immobilised life stages of the test species; GreenScreen EM, a yeast based assay for genotoxicity and general acute toxicity supplied by Gentronix Ltd; and CellSense a mediated, amperometric whole cell biosensor based on immobilised activated sludge and E. coli. 38 effluent samples supplied by members of SOCSA (Specialised Organic Chemicals Sector Association) were examined over a period of 13 months, in the project co-ordinated by the AstraZeneca Brixham Environmental Laboratory, and part funded by BioWise via the UK Government Department of Trade and Industry. PMID- 15536499 TI - Intervention study of airborne fungal spora in homes with portable HEPA filtration units. AB - The concentrations and composition of airborne fungal spores in homes fitted with portable HEPA filtration units were examined to provide information to evaluate the importance of varying levels of fungal spores in residential environments in Perth, Australia. A novel method for simulating activity/impaction on carpeted environments was also investigated. Reductions in fungal (35%) and particulate (38%) levels were achieved in the air filter homes. Penicillium, Cladosporium and yeasts were the most common and widespread fungi recovered indoors and outdoors. Fungal range decreased over the study period but this could be due to an overall reduced dissemination of spores (less spores in the air). PMID- 15536501 TI - A positive chemical ionization GC/MS method for the determination of airborne ethylene glycol and propylene glycols in non-occupational environments. AB - An analytical method for ethylene glycol and propylene glycols has been developed for measuring airborne levels of these chemicals in non-occupational environments such as residences and office buildings. The analytes were collected on charcoal tubes, solvent extracted, and analyzed by gas chromatography-mass spectrometry using a positive chemical ionization technique. The method had a method detection limit of 0.07 microg m(-3) for ethylene glycol and 0.03 microg m(-3) for 1,2- and 1,3-propylene glycols, respectively, based on a 1.44 m3 sampling volume. Indoor air samples of several residential homes and other indoor environments have been analyzed. The median concentrations of ethylene glycol and 1,2-propylene glycol in nine residential indoor air samples were 53 microg m(-3) and 13 microg m(-3) respectively with maximum values of 223 microg m(-3) and 25 microg m(-3) detected for ethylene glycol and 1,2-propylene glycol respectively. The concentrations of these two chemicals in one office and two laboratories were at low microg m(-3) levels. The maximum concentration of 1,3-propylene glycol detected in indoor air was 0.1 microg m(-3). PMID- 15536500 TI - Personal air sampling and analysis of polybrominated diphenyl ethers and other bromine containing compounds at an electronic recycling facility in Sweden. AB - Brominated flame retardants (BFRs) have been used extensively and are considered ubiquitous contaminants. To evaluate exposure to brominated flame retardants within an electronic recycling facility personal air monitoring was done during a two year period. A total of 22 polybrominated diphenyl ethers (PBDEs) and 2 other bromine containing organic compounds have been analysed and evaluated in 17 personal air samples. The most abundant congeners of PBDE was #209 (<0.7-61 ng m( 3)), #183 (<0.1-32 ng m(-3)) indicating the use of the commercial octaBDE mixture, followed by PBDE #99 and #47 (<1.3-25 and <0.9-16 ng m(-3), respectively). The second most abundant peak in the chromatogram from all samples was identified as 1,2-bis(2,4,6-tribromophenyxy)ethane (BTBPE) in the concentration range <0.6-39 ng m(-3) (semi-quantitatively calculated against PBDE #191). A second bromine containing compound was also detected, structurally similar to decabromodiphenyl ethane (DeBDethane), however no definite identification could be made. The air samples were also evaluated on a work exposure category basis. The workers represented three different categories: dismantlers, other workers and unexposed. There was a significant difference (p < 0.05 with the Mann-Whitney test) among the dismantlers and the unexposed categories for PBDE congeners #47, #100, #99, #154; #153, #183, #209 and BTBPE. Another observation was that the air concentrations of PBDEs and BTBPE in the breathing zone were negatively correlated (p < 0.05) to the amount of recycled material (in kg). The present work shows that the exposure to brominated flame retardants varied within the electronic facility and that further research is needed to evaluate how the exposure differs with different products being dismantled as well as how the bioavailability of the different BFRs to humans is related to particle exposure. PMID- 15536502 TI - Effects of soil copper and nickel on survival and growth of Scots pine. AB - The contribution of soil Cu and Ni pollution to the poor vitality and growth rate of Scots pine growing in the vicinity of a Cu-Ni smelter was investigated in two manipulation experiments. In the first manipulation, Cu-Ni smelter-polluted soil cores were transported from a smelter-pollution gradient to unpolluted greenhouse conditions. A 4-year-old pine seedling was planted in each core and cultivated for a 17-month period. In the second manipulation, pine seedlings from the same lot were cultivated for the same 17-month period in a quartz sand medium containing increasing doses of copper sulfate, nickel sulfate, and a combination of both. The variation in the biomass growth of the seedlings grown in the smelter-polluted soil cores was very similar to that of mature pine stands growing along the same smelter-pollution gradient in the field. In addition, the rate of Cu and Ni exposure explained a high proportion of the biomass growth variation, and had an effect on the Ca, K, and Mg status of the seedlings. According to the lethal threshold values determined on the basis of the metal sulfate exposure experiments, both the Cu and Ni content of the 0.5 km smelter polluted soil cores were high enough to cause the death of most of the seedlings. The presence of Cu seemed to increase Ni toxicity. PMID- 15536504 TI - Total mercury determination in sand boxes from Montreal. AB - Direct mercury analysis was successfully applied to determine trace levels of total mercury in samples from sand boxes in Montreal (Quebec, Canada). Twenty sand boxes were sampled from across the city and divided into two size fractions, a fine fraction (<100 microm) and a whole fraction. The concentrations of mercury ranged from 1.6 to 35 microg Hg kg(-1) dry soil for the fine fraction and from 0.7 to 6 microg Hg kg(-1) dry soil for the whole fraction. The mercury concentrations correlated with the soil organic carbon content (R2= 0.67) in the sand. The ratio of the concentration of mercury in the fine over the whole fraction varied from 2.2 to 18. Using published soil ingestion rates for children, the calculated daily intake values varied from 0 to 0.5 ng Hg kg(-1) bw d(-1) with an estimated oral ingestion of 200 mg of sand and from 0.2 to 4.7 ng Hg kg(-1) bw d(-1) with an ingestion of 1750 mg of sand. None of the sand boxes contain sufficient amounts of mercury so as to exceed the currently accepted daily intake threshold of 0.105 microg Hg kg(-1) bw d(-1) established by Health Canada. PMID- 15536503 TI - Simultaneous determination of 2-phenylbenzotriazole-type mutagens, PBTA-1 through -8, in river water by liquid chromatography-tandem mass spectrometry. AB - We describe a method for the simultaneous determination of eight kinds of phenylbenzotriazole-type mutagens (PBTA-1, -2, -3, -4, -5, -6, -7 and -8) in river water based on liquid chromatography-tandem mass spectrometry (LC/MS/MS). The application of dopant-assisted atmospheric pressure photoionization (APPI) for the detection of the PBTAs was studied. The APPI technique provided higher PBTA signal intensities than those obtained with an electrospray ionization (ESI) source, and the APPI method was used for the determination of the PBTAs. A solid phase extraction procedure was used for the extractions of PBTA-1 through -8 from river water. The procedure was rapid and the relative standard deviations were below 15%. The detection limits of PBTA-1 through -8 in river water using the proposed method were found to range from 0.04 to 0.5 ng L(-1) and PBTAs were successfully detected in river water at sub-ng L(-1) levels. PMID- 15536505 TI - The impact of a disused mine on uranium transport in the River Fal, South West England. AB - Unfiltered and filtered (0.45 and 0.2 microm) water samples and sediment samples (sieved to <180 microm and 180-1000 microm) were collected along an approximately 15 km transect of the River Fal, Cornwall, UK, to examine the impact of the disused South Terras uranium mine on the uranium concentrations of the river water and underlying sediments. The uranium concentration of the water samples fluctuated along the river, with the 0.45 microm filtered water showing the largest, seven-fold, difference between minimum (0.19 microg L(-1)) and maximum (1.34 microg L(-1)) concentrations. The historical uranium mine and spoil heaps were not a significant source of uranium to the river water, as water concentrations were low next to the site, but a highly elevated uranium concentration (1000 mg kg(-1)) was found in sediment below an outflow pipe from this mine. Operationally defined "colloidal" (0.2-0.45 microm) and "dissolved" (<0.2 microm) uranium were the predominant forms of the element in the river water (35 and 45% respectively). The uranium concentration in the dissolved phase showed a correlation coefficient of 0.83 (n= 9) with the total cation concentration, suggesting that the uranium concentration in this fraction is directly linked to weathering of rocks and minerals. The observation that weathering is the dominant mechanism delivering uranium to the river water explains the low uranium concentrations in the river water close to South Terras mine, despite the proximity of the spoil heaps, and the maximum uranium concentrations close to a china clay mining area. PMID- 15536506 TI - PBDEs in marine and freshwater sediments from Belgium: levels, profiles and relations with biota. AB - Sediments from the Belgian North Sea (BNS), the Western Scheldt Estuary (SE) and freshwater watercourses from the Scheldt basin were analysed for eight PBDE congeners, namely BDEs 28, 47, 99, 100, 153, 154, 183 and 209. Previously analysed biological samples from the same locations in the BNS and the SE have been shown to contain large amounts of PBDEs. Surprisingly, PBDE concentrations in the sediments were below the LOQ for samples from the BNS (except BDE 209), while in those from the SE the sum of PBDEs (not including BDE 209) were higher and ranged from 0.20 to 0.41 ng g(-1) dw. BDE 209 could be detected in 83% of the samples from the BNS and in all the samples from the SE. Concentrations up to 1200 ng g(-1) were hereby measured in the SE. Compared to the marine and estuarine locations, the sediments from the freshwater watercourses were relatively more polluted with the lower brominated PBDEs (<0.20-19 ng g(-1) dw). BDE 209 concentrations up to 320 ng g(-1) dw were measured in those sediments. However, the contribution of BDE 209 to the total amount of PBDEs varied much more at the freshwater locations than in the SE, which suggests a different input of pollutants. PBDE profiles observed in biological samples do not match the profiles of the sediments. BDE 183 and 209 could not be quantified in biota, although these congeners were undoubtedly present in the sediments. This raises questions about the bioavailability of these congeners in the environment. PMID- 15536507 TI - Field uptake rates of hydrophobic organic contaminants by semipermeable membrane devices: environmental monitoring considerations. AB - The uptake rates of selected hydrophobic organic contaminants (HOCs) by semipermeable membrane devices (SPMDs)--a polyethylene layflat containing the lipid triolein--were investigated under natural conditions. SPMDs were exposed in three sampling sites (industrial, urban, and agricultural areas) in the Tajo River (Toledo, Spain) for 5, 11 and 20 d. The organochlorine compounds 4,4'-DDT, 4,4'-DDE, alpha-HCH, gamma-HCH, pentachlorobenzene, hexachlorobenzene, and polychlorinated biphenyls (PCBs), and the 16 priority pollutant polycyclic aromatic hydrocarbons (PAHs) were detected in the SPMDs deployed in the three sampling sites. A linear uptake rate was found for DDTs and for 4-Cl- and 5-Cl substituted PCB congeners in all sampling sites. Concentrations of HCHs (80.3 ng g(-1) SPMD for alpha-HCH and 109 ng g(-1) SPMD for gamma-HCH after 20 d of exposure) increased according to a linear uptake rate in the SPMDs deployed in the sampling site located in the agricultural area. Likewise, a marked increase of total PAH concentration (up to 300 ng g(-1) SPMD after 20 d of exposure) was solely found in the sampling site situated near a thermoelectric power station. Examination of individual PAHs revealed that PAHs with log K(OW) between 4.2 and 5.7 displayed a linear uptake rate over the 20 d of exposure. Water concentrations (ng L(-1)) of HCB (0.80-2.48), lindane (1.30-11.5), 4,4'-DDT (0.61 2.02), 4,4'-DDE (6.89-11.6) and total PAHs (12.0-26.7) estimated by a linear uptake kinetic model were found to be high in comparison with other polluted aquatic systems, and similar to concentrations in other Spanish rivers. Our results suggest that SPMD kinetic uptake studies in the natural environment are recommended for identifying point-pollution sources, and that shorter times of SPMD exposure (approximately 1 week) are desirable to minimize one of the main problems of field SPMD deployment, i.e., the biofouling, which negatively affects the estimation of the dissolved HOC concentrations. PMID- 15536508 TI - Probabilistic human health risk of PCDD/F exposure: a socioeconomic assessment. AB - The aim of the present study was to assess the human health risks (carcinogenic and non-carcinogenic) of PCDD/F exposure for the adult population living in two different areas (industrial and residential) of Tarragona (Catalonia, Spain), as well as to compare these risks according to the socioeconomic status of the population. Two pathways were considered for PCDD/F exposure: direct (air inhalation, dermal contact and ingestion from soils and dust) and dietary. Although environmental exposure was higher for the population living in the neighborhood of the industrial area (mainly lower socioeconomic group subjects), it only accounted 1.05%. On the other hand, PCDD/F exposure through dietary intake was more important for those individuals of an upper socioeconomic level. This was due to the fact that their diet is richer in those foodstuffs containing higher PCDD/F concentrations. Consequently, taking together direct and dietary exposure to PCDD/Fs, the result is that upper socioeconomic group populations are more exposed to these toxic pollutants. In any case, the current levels of PCDD/F exposure would not mean significant carcinogenic and non-carcinogenic risks for the population living in the industrial and residential areas of Tarragona. PMID- 15536509 TI - Peak exposures in aluminium potrooms: instrument development and field calibration. AB - Aluminium smelter potrooms are unique in that workplace exposures to hydrogen fluoride (HF), sulfur dioxide (SO2), and particulate matter occur simultaneously for some tasks. The peak exposures to these contaminants are of increasing interest in discovering the etiology of respiratory health effects. While a variety of direct-reading instruments are available for sulfur dioxide and particulate matter, only a few exist for hydrogen fluoride. The sensors in these HF instruments have a cross-sensitivity to sulfur dioxide making it difficult to monitor HF in an environment that also contains SO2. To overcome this problem, we assessed the simultaneous use of two electrochemical instruments: one with a SO2 sensor that does not respond to HF and the second with a hydrogen fluoride sensor that responds to both HF and SO2 in a 1 : 1 ratio, termed 'total acid gas'. The difference in the response between the two instruments should indicate the HF concentration: [HF + SO2] minus SO2 equals HF. The performance characteristics of this sampling train were evaluated in the laboratory through the generation of both HF and SO2 with permeation tubes. The response and recovery times for the SO2 only instrument were acceptable (6 and 15 s, respectively), but the "total acid gas" instrument exhibited both slow response and slow recovery approaching three and six min. The association between the traditional integrated filter sampling method and the direct-reading instrument for SO2 is 0.80 (Spearman's rho). The use of the digital filter strengthens the association between the HF direct-reading instrument and the integrated samples from 0.41 to 0.68. PMID- 15536510 TI - New horizons in molecular informatics. PMID- 15536512 TI - [Vascular injuries in extremities]. AB - Vascular injuries of the extremities account for most instances of vascular trauma (ca. 70%), and they entail a risk of amputation about 10-20%. According to the kind of force that has acted, arterial trauma is classified as direct or indirect. The scale of hemorrhage and peripheral ischemia depend on the nature and severity of the arterial lesion. In patients with multiple injuries, routine use of Doppler sonography and duplex sonography can facilitate early diagnosis and treatment of vascular injuries. With great certainty, clinical examination and an AB or WB index of >1.0 can rule out the presence of vascular injury that requires treatment. After excluding further life-threatening injuries, surgery should be performed immediately when there is critical ischemia, squirting hemorrhage, or a rapidly expanding hematoma. Angiography or duplex sonography findings determine the further procedure in vascular injuries that do not require immediate treatment. Occlusion of a reconstructed artery, manifestation of a compartment syndrome, and insufficient anticoagulation are the main factors affecting the risk of amputation. PMID- 15536515 TI - [Pruritus in pregnancy. A frequent diagnostic and therapeutic challenge]. AB - Pruritus is the leading dermatological symptom during pregnancy. Besides preexisting or acquired dermatoses, there are a number of pregnancy-specific dermatological diseases such as PEP (polymorphic eruption of pregnancy, previously named PUPPP), pemphigoid (herpes) gestationis, and pruritus gravidarum that are accompanied by severe itching and scratching. Because of potential effects on the fetus, the treatment of pruritus in pregnancy requires prudent consideration. The use of topical and systemic treatments depends on the underlying aetiology of pruritus and the stage and status of the skin. In general, emollients, topical anti-pruritics and topical corticosteroids appear to be the safest options for localised forms of pruritus in pregnancy whereas systemic treatments and/or UV phototherapy are adequate for generalized pruritus. Systemic corticosteroids and a restricted number of antihistamines may be administered in severe cases. This paper highlights the major aetiologies of pruritus during pregnancy and points out the cornerstones of antipruritic therapy in recognition of our own clinical experiences and the current literature. PMID- 15536516 TI - [Oral diabetes treatment. Which substance is indicated at which time?]. AB - The prevalence of type 2 diabetes continues to show a clear upward trend in Germany. In earlier days it was considered the "harmless diabetes of old age," but has become increasingly recognized as a disease carrying a high risk of vascular sequelae as well as shortening the diabetic's remaining life expectancy if adequate therapy is not initiated. In addition to correcting hyperglycemia, treatment consists in effective management of concomitant risk factors such as hypertension, dyslipidemia, and adiposity resulting from faulty nutrition and lack of exercise. In the large majority of overweight type 2 diabetics, metformin is the oral antidiabetic agent of first choice provided the patient does not exhibit renal insufficiency, which represents the most important contraindication. This recommendation for monotherapy of overweight type 2 diabetics is supported by an endpoint study. In contrast, no equivalent evidence is available on any of the possible options for oral combination therapy. PMID- 15536517 TI - [Shaping the future of diabetic medicine together]. PMID- 15536518 TI - Antisense oligonucleotides and short interfering RNAs silencing the cyclin dependent kinase inhibitor p21 improve proliferation of Duchenne muscular dystrophy patients' primary skeletal myoblasts. AB - Increased levels of the cyclin-dependent kinase inhibitor p21 associated with decreased myoblast proliferation may be involved in the dystrophic process in Duchenne muscular dystrophy (DMD). Therefore we are interested to improve the proliferation of primary myoblasts of DMD patients by a reduction in p21 using either antisense oligonucleotides (ASO) or short interfering RNAs (siRNA). After transient transfection of myoblasts in cell culture proliferation was analyzed using a 5-bromo-2'-deoxyuridine assay comparing specific transfected cells with untransfected cells and cells transfected with scrambled ASO and luciferase siRNA, respectively. Four of five Dystrophin-deficient (Dys(-)) cell culture samples revealed an increase in proliferation between 7% and 18% compared to untransfected cells and between 8% and 36% compared to cells transfected with scrambled ASO. Transfection with siRNA was performed for selected samples to determine whether siRNA is more effective in gene silencing than ASO. The increase in proliferation using luciferase siRNA as reference was comparable to or less than ASO data using scrambled ASO as reference. Using untransfected cells as reference, the increase in proliferation was higher for siRNA than ASO (20-47% vs. 7-18%), but the data must be carefully interpreted with respect to nonspecific effects on gene expression by siRNA. Our findings of transient p21 gene silencing represent a basis for viral vector-mediated drug-inducible p21 shRNA expression in Dys(-) myoblasts which might enhance, prolong and regulate the proliferation effect. PMID- 15536519 TI - Tamoxifen-resistant breast cancers show less frequent methylation of the estrogen receptor beta but not the estrogen receptor alpha gene. AB - No reports have examined the association between tamoxifen resistance and the methylation of the estrogen receptor (ER) alpha and beta genes. Therefore we investigated the methylation patterns of the ER genes in the tamoxifen-resistant tumors. We used bisulfite genomic sequencing and reverse transcriptase PCR to determine the methylation patterns and mRNA expression of the two ER genes from control (n = 68) and tamoxifen-resistant tissues (n = 34) chosen by an age matched sampling method. Bisulfite genomic sequencing allowed us to reveal the methylation of the ER alpha gene in 15 of the control tumors (22.1%) and in 11 tumors of the resistant group (32.4%). The methylation of ER beta was observed in 40 control tumors (58.8%) and in 8 recurrent tumors (23.5%). The methylation rate of the ER beta but not the ER alpha in the control group was significantly higher than in its counterpart (ER alpha, P = 0.261; ER beta, P = 0.001). Among the methylated tumors mean methylation density of ER alpha and ER beta in the resistant cases was significantly elevated (ER alpha, P = 0.014; ER beta, P < 0.001). Furthermore, the expression rate of ER beta mRNA was higher among the tumor in the resistant group than in the control with marginal significance (77.8% vs. 38.1%, P = 0.109). Additionally, in the cancers from the resistant cases, the cells showed a higher percentage of positive staining for Ki67 than those from the control group (P = 0.001). Our study indicates that there is an inverse relationship between the methylation rate of the ER beta gene and tamoxifen resistance. The tamoxifen-resistant tumors showed more dense methylation of the ER beta gene than control tumors. Although the number of case samples was limited, our results support the hypothesis that hypermethylation of the ER beta gene negatively affects the development of tamoxifen resistance. PMID- 15536520 TI - Association of polymorphisms of the androgen receptor and klotho genes with bone mineral density in Japanese women. AB - Genetic variants of the androgen receptor and klotho protein may contribute to variation in bone mass as well as to predisposition to osteoporosis. The relationship of a CAG repeat polymorphism of the androgen receptor gene (AR) and of a -395G-->A polymorphism of the klotho gene (KL) to bone mineral density (BMD) in Japanese women was examined in a population-based study. The subjects (1,101 and 1,110 women for AR and KL polymorphisms, respectively) were aged 40-79 years and were randomly recruited to a population-based prospective cohort study of aging and age-related diseases. BMD for the total body, lumbar spine, right femoral neck, right trochanter, and right Ward's triangle was measured by dual energy X-ray absorptiometry. Genotypes for the AR and KL polymorphisms were determined by polymerase chain reaction based assays. The number of CAG repeats of AR was inversely correlated with BMD for the lumbar spine in premenopausal women but not in postmenopausal women. The (CAG)(n/=23) alleles were designated S and L, respectively. Among premenopausal women, BMD for the total body was significantly lower in subjects with the LL genotype than in those with the SS genotype or those in the combined group of SS and SL genotypes. In contrast, BMD was not associated with AR genotype in postmenopausal women. Among all women, BMD for the lumbar spine was significantly lower in subjects with the GG genotype of the -395G-->A polymorphism of KL than in those with the AA genotype. BMD was not associated with -395G-->A genotype among premenopausal women. In postmenopausal women, BMD for the total body or lumbar spine tended to be lower in subjects with the GG genotype than in those with the AA genotype or those in the combined group of GA and AA genotypes. These results suggest that AR is a susceptibility gene for reduced BMD in premenopausal Japanese women, and that KL is a susceptibility gene for reduced BMD in all women. PMID- 15536521 TI - The first haramiyoid mammal from Asia. AB - A lower molar of a haramiyoid mammal is described from the Toutunhe Formation of Liuhuanggou near Urumqi in the southern Junggar Basin, Xinjiang, China. It is referred to Eleutherodon sp., otherwise exclusively known from the Upper Bathonian of England. It is the first record of the order Haramiyida from Asia and the first Mesozoic mammal described from the southern Junggar Basin. Apart from the English specimens of Eleutherodon and Staffia from the Upper Jurassic of East Africa, it is the geologically youngest haramiyoid known. It is the first vertebrate taxon from the Toutunhe Formation that is probably not endemic and lends some support to the dating of the Formation as late Middle Jurassic, probably Bathonian. PMID- 15536522 TI - [The syndrome of genital retraction from a transcultural psychiatric point of view. Chinese suo yang, Indonesian koro and non-Asian forms (koro-like symptoms)]. AB - The present article reviews the syndrome of genital retraction (SGR) from the perspective of transcultural psychiatry. It is best known as a culture-bound syndrome in Asia, e. g. koro in Indonesia or suo yang in Southern China, where it can be also observed in so-called epidemics. The syndrome is characterised by the sensation that the genitals are shrinking and being pulled back into the abdomen; this perception is associated with fear of death, because the subjects expect to die as soon as the genital has disappeared completely. Particularly in the case of suo yang, the involvement of local traditional concepts of illness can be established. In the Western world, however, SGR is observed only in single cases. The symptoms are less dramatic and do not show a particular cultural context. This form of SGR is called "koro-like" (koro-like symptoms, KLS). In contrast to the Asian form, KLS are not observed in healthy subjects, but as an unspecific syndrome related to other neuropsychiatric illnesses. In this review, we compare both forms of the SGR in terms of symptoms, cultural aspects and therapeutic strategies. PMID- 15536524 TI - Length of weaning from mechanical ventilation. PMID- 15536523 TI - Genetic diversity associated with variation in silage corn digestibility for three O-methyltransferase genes involved in lignin biosynthesis. AB - Polymorphisms within three candidate genes for lignin biosynthesis were investigated to identify alleles useful for the improvement of maize digestibility. The allelic diversity of two caffeoyl-CoA 3-O-methyltransferase genes, CCoAOMT2 and CCoAOMT1, as well as that of the aldehyde O-methyltransferase gene, AldOMT, was evaluated for 34 maize lines chosen for their varying degrees of cell wall digestibility. Frequency of nucleotide changes averaged one SNP every 35 bp. Ninety-one indels were identified in non-coding regions and only four in coding regions. Numerous distinct and highly diverse haplotypes were identified at each locus. Numerous sites were in linkage disequilibrium that declined rapidly within a few hundred bases. For F4, an early flint French line with high cell wall digestibility, the CCoAOMT2 first exon presented many non synonymous polymorphisms. Notably we found an 18-bp indel, which resembled a microsatellite and was associated with cell wall digestibility variation. Additionally, the CCoAOMT2 gene co-localized with a QTL for cell wall digestibility and lignin content. Together, these results suggest that genetic diversity investigated on a broader genetic basis could contribute to the identification of favourable alleles to be used in the molecular breeding of elite maize germplasm. PMID- 15536525 TI - Measurement of the liver tissue oxygenation by near-infrared spectroscopy. AB - OBJECTIVE: To study the relation between the liver tissue oxygenation index (TOI), transcutaneously measured with spatially resolved spectroscopy (a new method of near-infrared spectroscopy or NIRS), the mixed venous oxygen saturation and the blood flow in the different parts of the splanchnic circulation in newborn piglets. DESIGN: Tissue oxygenation index of the liver was measured in six newborn piglets at 33 degrees C, 35 degrees C, 37 degrees C and after a decrease in arterial carbon dioxide pressure (PaCO(2)). MEASUREMENTS: Mixed venous oxygen saturation, blood gas analysis and peripheral oxygen saturation were measured at each step. Gastric, proximal jejunal, midgut, distal ileal, splenic and hepatic arterial blood flow were measured by injection of coloured microspheres into the left atrium. NIRS optodes were attached to the skin over the liver and TOI was calculated. RESULTS: No significant changes of TOI of the liver were seen during the increase in temperature or change in PaCO(2). TOI correlated well with mixed venous oxygen saturation (r=0.85), the mid-ileal blood flow (r=0.57) and the distal ileal blood flow (r=0.72). CONCLUSIONS: Measurement of the TOI of the liver might be a non-invasive way to measure the distal ileal blood flow. PMID- 15536526 TI - The neuroendocrine stress response and severity of acute respiratory syncytial virus bronchiolitis in infancy. AB - OBJECTIVE: Neuroendocrine hormones have profound effects on the immune system. The immune response is a major factor in the pathogenesis of acute respiratory syncytial virus (RSV) infection. We hypothesised that there is a relationship between the neuroendocrine response in acute RSV infection, the severity of illness, and the degree of lymphopenia. DESIGN: Prospective, non-randomised cohort study of infants hospitalised for RSV infection requiring mechanical ventilation or managed conservatively. The study assessed the effect of age, gender, birth gestation, and severity of illness on stress hormone profile and its relationship to lymphocyte count. SETTING: Regional Paediatric Intensive Care Unit (PICU) and children's wards. PATIENTS: Thirty-two consecutive infants with RSV infection were enrolled, of which thirteen were mechanically ventilated on PICU (study subjects) and nineteen treated on the ward (comparison group). Twenty three children (72%) returned for follow-up. MEASUREMENTS AND MAIN RESULTS: A specific neuroendocrine profile was found in PICU patients compared to ward patients (Wilks Lambda = 0.36, F = 9.05, P =.03). PICU patients had significantly higher prolactin and growth hormone, and significantly lower leptin and IGF-1. Cortisol levels were the same. PICU patients were more lymphopenic compared to ward patients (P =.0001). On multiple regression analysis, prolactin and leptin levels accounted for 57% of the variation in lymphocyte count. CONCLUSIONS: Whereas the effect of intensive care (mechanical ventilation and medication) could not be controlled for, our results suggest that there is an association between the neuroendocrine hormone response, severity of illness and degree of lymphopenia. PMID- 15536527 TI - Influence of positive end-expiratory pressure (PEEP) on histopathological and bacteriological aspects of pneumonia during low tidal volume mechanical ventilation. AB - OBJECTIVE: Ventilatory strategies combining low tidal volume (V(T)) with positive end-expiratory pressure (PEEP) are considered to be lung protective. The influence of the PEEP level was investigated on bacteriology and histology in a model of ventilator-associated pneumonia. SUBJECTS: Nineteen New Zealand rabbits. INTERVENTIONS: The animals were mechanically ventilated with a positive inspiratory pressure of 15 cmH(2)O and received either a zero end-expiratory pressure (ZEEP, n=6), a 5 cmH(2)O PEEP (n=5) or a 10 cmH(2)O PEEP (n=4). An inoculum of Enterobacter aerogenes was then instilled intrabronchially. The non ventilated pneumonia group (n=4) was composed of spontaneously breathing animals which received the same inoculum. Pneumonia was assessed 24 h later. MAIN RESULTS: The lung bacterial burden was higher in mechanically ventilated animals compared with spontaneously breathing animals. All animals from the latter group had negative spleen cultures. The spleen bacterial concentration was found to be lower in the 5 cmH(2)O PEEP group when compared to the ZEEP and 10 cmH(2)O PEEP groups (3.1+/-1.5 vs 4.9+/-1.1 and 5.0+/-1.3 log(10) cfu/g, respectively; p<0.05). Lung weight and histological score values were lower in the spontaneously breathing animals as well as in the 5 cmH(2)O PEEP group compared with the ZEEP and 10 cmH(2)O groups. CONCLUSIONS: Mechanical ventilation substantially increased the lung bacterial burden and worsened the histological aspects of pneumonia in this rabbit model. Variations in terms of lung injury and systemic spreading of infection were noted with respect to the ventilatory strategy. PMID- 15536528 TI - Antibiotic pharmacokinetic and pharmacodynamic considerations in critical illness. AB - BACKGROUND: Many factors over which there may be little control may influence the response of a patient to therapy. However, therapy with antibiotics can be readily optimised. DISCUSSION: Concentration-dependent agents such as aminoglycosides appear effective and to entail fewer side effects when given in large, infrequent doses. There is also evidence that time-dependent antibiotics often fail to reach adequate concentrations throughout the treatment period. To date no randomised controlled prospective trial has demonstrated improvement in clinical outcome following infusion rather than intermittent boluses of time dependent antibiotics. Critical illness alters antibiotic pharmacokinetics principally through increases in volume of distribution. Other than glycopeptides and aminoglycosides, antibiotic blood concentrations are rarely monitored and therefore adequate concentrations can only be inferred from clinical response. CONCLUSIONS: Failure to respond within the first few days of empirical treatment may be due to antibiotic resistance or inadequate doses. Therefore the same rigor should be applied to achieving adequate antibiotic concentrations as is applied to inotropes, which are titrated to achieve predetermined physiological targets. PMID- 15536535 TI - An osteoporosis screening tool for Chinese men. AB - Several osteoporosis risk instruments have been proposed to select women for bone densitometry, but no validated instruments are currently available for men. This study aims to address this deficiency by developing and validating a Male Osteoporosis Screening Tool (MOST) for Chinese men. Two thousand ambulatory men, aged 65 and above, were recruited from the general community in Hong Kong, and a cohort of 1,970 men with valid total hip and lumbar spine dual-energy X-ray absorptiometry (DXA) measurements was included in the current analysis. A 60% random sample was selected as the training sample for developing the screening tool, and the remaining 40% constituted the validation sample. Logistic regression and receiver operating characteristic (ROC) analysis were used to identify the simplest combination of risk factors to be included in the screening tool for predicting osteoporosis at the femoral neck, total hip, or lumbar spine. Body weight and quantitative ultrasound index (QUI) were found to contribute significantly to the area under the ROC curve (AUC), yielding an AUC of 0.823 in the training sample. The resulting MOST had a sensitivity of 94% and a specificity of 46% when using a cutoff score of 3. MOST had an AUC of 0.839 in the validation sample. The risk of osteoporosis was 1% among those with MOST scores < or = 2, but 72% among those with MOST scores > 7. Using a cutoff of 3, the negative predictive value was 97.5% which suggests that the 42% with MOST scores < or = 3 may be accurately screened out as being without osteoporosis, thus saving two fifths of our DXA resources. The positive predictive value was 72% when using a cutoff of 7, implying that MOST cannot replace DXA for case finding purposes. Nevertheless, for resource allocation and patient satisfaction, it is prudent and economical to offer DXA screening first to the 6% with MOST scores > 7. PMID- 15536536 TI - The challenge: fracture treatment in osteoporotic bone. PMID- 15536537 TI - New implant designs for fracture fixation in osteoporotic bone. AB - Screws are one of the limiting factors for fixation of implants, particularly in poor bone quality. A class of new implants with an implant-bone-interface optimized regarding load transition by increasing the peripheral area might improve the anchorage of implants in osteoporotic bone. However, the shape of these implants requires new technologies for insertion. The goal of the work presented here was to analyze the relevant parameters regarding implant geometry and to demonstrate the effect of new procedures for their insertion. The investigation was divided into three parts: 1) implant design optimisation, 2) efficiency of cortical bone ablation, and 3) implant insertion technology. Finite element analysis (FEA) was performed to investigate the influence of the number of lobes, the radius of the outer curvature and additional milling to remove any sharp changes of section around the lobe. Opening of the cortical bone with an Er:YAG laser was studied using calf cortex from 2 to 7 mm thickness. The effect of a) pulse energy and pulse duration, b) cortical thickness, c) wet or dry boundary conditions on volume and geometry of ablated bone, time required to penetrate the cortical bone and local bone tissue damage was quantified. Pneumatic and ultrasound based insertion were compared in the third experiment. The cortical bone was prepared in the following ways: a) no opening, b) predrilling of three holes (1 mm diameter each) and c) exact pre-cutting of the whole contour. Increasing the radius of the outer curvature from 2 to 5 mm reduces the peak stresses during loading in all planes in the implant as well as in the adjacent cortical bone by about 30-40%. An increase in the number of lobes from two to three decreases the mean peak stress by about 46% (alpha < 0.001) and the range between the minimal and maximal peak stresses for different loading directions by about 83%. Penetration of cortical bone with an Er:YAG laser was possible up to a cortical thickness of 6 mm with fewer than 100 pulses. The ablation rate per pulse increased more with increasing duration than with increasing energy. Signs of bone damage such as melting were only visible when high pulse energies and durations were used. Insertion of the prototype was possible with all devices, but only when the whole contour was cut out of the cortical bone. However, the use of the ultrasound vibrator led to heating up of the tissue fluid and subsequently to water evaporation and tissue damage. Insertion of the prototype was possible with both pneumatic vibrators, but only when the whole contour was cut out of the cortical bone. New implant designs may lead to reduced stress peaks in the surrounding bone and might be inserted with the help of new insertion technologies, namely laser cutting of cortical bone and pneumatic vibration. Further studies are required to optimize these technologies prior to clinical use. PMID- 15536538 TI - Fixation principles in metaphyseal bone--a patent based review. AB - Osteoporotic changes start in cancellous bone due to the underlying pathophysiology. Consequently, the metaphyses are at a higher risk of "osteoporotic" fracture than the diaphysis. Furthermore, implant purchase to fix these fractures is also affected by the poor bone quality. In general, researchers and developers have worked on three different approaches to address the problem of fractures to osteoporotic bone: adapted anchoring techniques, improved load distribution as well as transfer with angular stable screws, and augmentation techniques using bone substitutes. A patent-based review was performed to evaluate which ideas were utilized to improve fixation in osteoporotic, metaphyseal bone, especially in the proximal femur, and to analyze whether the concept had entered clinical use. Anchoring devices that are either extramedullary or intramedullary have a long clinical history. However, demanding surgical techniques and complications, especially in poor quality bone, are justification that such implants and their corresponding surgical techniques need to be improved upon. Expanding elements have been evaluated in the laboratory. The results are promising and the potential of this approach has yet to be fully exploited in the clinics. Internal fixators with angular stable screws open the door for many new anchorage ideas and have great potential for further optimization of load distribution and transfer. Augmentation techniques may improve anchorage in osteoporotic bone. However, the properties of bone substitute materials will need to be modified and improved upon in order to meet the demanding requirements. If we summarise the development process and the clinical use of implants to date, we have to clearly state that more factors than simply biomechanical advantage will determine the clinical success of a new fixation principle or a new implant. Instead, fracture treatment of patients with osteoporosis really needs an interdisciplinary approach! PMID- 15536539 TI - Physical training and hormone replacement therapy reduce the decrease in bone mineral density in perimenopausal women: a pilot study. AB - The effects of physical training and hormone replacement therapy (HRT) on bone mineral density in perimenopausal women were studied. Sixty perimenopausal women were randomized to either physical training (n = 20), HRT (n = 20), or control group (n = 20). The study period was 18 months. Bone mineral density (BMD) in the femoral neck and lumbar spine was measured using dual-energy X-ray absorptiometry (DXA). DXA was performed before treatment and after 6 and 18 months. Blood samples for analysis of the bone markers U-deoxypyridinoline and osteocalcin were collected at the same time points. After 18 months, BMD in the spine had not decreased in either the training group or in the HRT group. In the control group, spine BMD had significantly decreased (p = 0.0014). U-Deoxypyridinoline and osteocalcin were increased significantly in the control group (p = 0.0198, p = 0.0295, respectively). No significant changes in bone marker levels were found in the training group or the HRT group. We found that both HRT and physical training can prevent loss of spine BMD in perimenopausal women over a period of 18 months. HRT remains a cornerstone in the treatment of vasomotor symptoms and preservation of BMD. However, HRT can only be used for limited periods of time due to the potential serious adverse effects. This study indicates a beneficial effect of physical activity on spine BMD in the perimenopausal period, and highlights its potential as an alternative to HRT during this period. PMID- 15536540 TI - Appropriateness of internal digital phantoms for monitoring the stability of the UBIS 5000 quantitative ultrasound device in clinical trials. AB - In bone status assessment, proper quality assurance/quality control is crucial since changes due to disease or therapeutic treatment are very small, in the order of 2-5%. Unlike for dual X-ray absorptiometry, quality control procedures have not been extensively developed and validated for quantitative ultrasound technology, limiting its use in longitudinal monitoring. While the challenge of developing an ideal anthropometric phantom is still open, some manufacturers use the concept of the internal digital phantom mimicking human characteristics to check the stability of their device. The objective of the study was to develop a sensitive model of quality control suitable for the correction of QUS patient data. In order to achieve this goal, we simulated a longitudinal device lifetime with both correct and malfunctioning behaviors. Then, we verified the efficiency of digital phantoms in detecting those changes and subsequently established the in vitro/in vivo relationship. This is the first time that an attempt to validate an internal digital phantom has made, and that this type of validation approach is used. The digital phantom (DP) was designed to mimic normal bone (BUAP2) and osteoporotic bone (BUAP1) properties. The DP was studied using the UBIS 5000 ultrasound device (DMS, France). Diverse malfunctions of the UBIS-5000 were simulated. Several series of measurements were performed on both BUAP1 and 2 and on 12 volunteers at each grade of malfunction. The effect of each simulated malfunction on in vivo and in vitro results was presented graphically by plotting the average BUA values against the percentage change from baseline. The change from baseline in BUA was modeled using linear regression, and the in vivo/in vitro ratio was obtained from the model. All experimentations influenced the measure of BUAP1 and 2 as well as the measure of our 12 volunteers. However, the degree of significance varied as a function of the severity of the malfunction, and the results also differed substantially in magnitude between in vivo and in vitro. Indeed, the DP was about 10 times more sensitive to variations of the transfer function than was the in vivo measurement, which is very reassuring. The sensitivity of the digital phantoms was reliable in the determination of simulated malfunctions of the UBIS-5000. The digital phantoms provided an accurate evaluation of the acoustic performance of the scanner, including the fidelity of transducers. In light of these results, the QC approach of the UBIS 5000 will be extremely simple to implement compared with other devices. Indeed, since the digital phantom was automatically measured during every patient measurement, the QC approach could be built on an individual level basis rather than on an average basis. PMID- 15536542 TI - Phosphorylation of c-Jun N-terminal Kinases (JNKs) is involved in the preventive effect of xanthorrhizol on cisplatin-induced hepatotoxicity. AB - Cisplatin is a potent anti-cancer chemotherapeutic agent but has the undesirable side effect of hepatotoxicity at high doses. In a previous study, abrogation of cisplatin-induced hepatotoxicity by pretreatment with xanthorrhizol was observed in mice, but the mechanism has not yet been studied. We therefore investigated whether the protective effect of xanthorrhizol on cisplatin-induced hepatotoxicity is associated with the mitogen-activated protein (MAP) kinase signaling pathway. Cisplatin caused phosphorylation of both c-Jun N-terminal kinases 1/2 (JNK1/2) and the extracellular signal-regulated kinase 1/2 (ERK1/2), but not that of p38. However, cisplatin-induced phosphorylation of JNKs, especially JNK1, was highly attenuated by pretreatment with xanthorrhizol in a dose-dependent manner. This study suggested that the phosphorylation of JNKs could be involved in the protective effect of xanthorrhizol on cisplatin-induced hepatotoxicity and it also affects gene transcription by regulating the expression of transcription factor subunits such as c-fos and p50 in part. In addition, considering that the expression of both cytochrome c and caspase-9 were not changed in this model, its mechanism might be independent of mitochondria related apoptosis. This is the first report giving evidence that the physiological function of xanthorrhizol is linked to regulation of the phosphorylation of JNK(s). PMID- 15536543 TI - Mutagenicity of the mycotoxin patulin in cultured Chinese hamster V79 cells, and its modulation by intracellular glutathione. AB - Because the ability of the mycotoxin patulin (PAT) to cause gene mutations in mammalian cells is still ambiguous, we have studied the mutagenicity of PAT at the hypoxanthine-guanine phosphoribosyltransferase (HPRT) gene locus in cultured Chinese hamster V79 cells with normal, depleted, and elevated glutathione (GSH) levels. PAT was more toxic to GSH-depleted cells than to normal cells and caused an increase of the intracellular GSH level in normal and GSH-depleted cells. It also caused synchronization of the cell cycle due to a temporary accumulation of cells in the G2/M phase; this G2/M arrest was more persistent in GSH-depleted than in normal cells. PAT gave rise to a clear and concentration-dependent induction of HPRT mutations at non-cytotoxic concentrations in V79 cells with normal GSH level; the lowest PAT concentration causing a significant number of mutant cells was 0.3 micromolar, and the mutagenic potency of PAT equaled that of the established mutagen 4-nitroquinoline-N-oxide. The mutagenicity of PAT was again more pronounced, by a factor of about three, in GSH-depleted V79 cells. Elevated GSH levels abolished all observed effects of PAT. These data support the notion that PAT is a mutagenic mycotoxin, in particular in cells with low GSH concentration. The ability of PAT to cause gene mutations in mammalian cells might have a bearing on its carcinogenicity. PMID- 15536544 TI - Olfactory mucosal toxicity screening and multivariate QSAR modeling for chlorinated benzene derivatives. AB - The olfactory mucosa (OM) is an important target for metabolism-dependent toxicity of drugs and chemicals. Several OM toxicants share a 2,6-dichlorinated benzene structure. The herbicides dichlobenil (2,6-dichlorobenzonitrile) and chlorthiamide (2,6-dichlorothiobenzamide) and the environmental dichlobenil metabolite 2,6-dichlorobenzamide all induce toxicity in the OM following covalent binding in the Bowman's glands. In addition, we have shown that 2,6 dichlorophenyl methylsulfone targets the Bowman's glands and is probably the most potent OM toxicant so far described. These findings suggest that the 2,6 positioning of chlorines in combination with an electron-withdrawing group in the primary position of the benzene ring is an arrangement that facilitates OM toxicity. This study examined the physicochemical characteristics of the 2,6 dichlorinated OM toxicants. A number of 2,6-dichlorinated benzene derivatives with various types of substituents in primary position were tested for OM toxicity in mice. In addition, some other 2,6- and 2,5-substituted benzene derivatives were examined. Two novel OM toxicants, 2,6-dichlorobenzaldehyde oxime and 2,6-dichloronitrobenzene, were identified. By the use of partial least squares projection to latent structures with discriminant analysis (PLS-DA) a preliminary quantitative structure-activity relationship (QSAR) model was built also using reported OM toxicity data. Physicochemical properties positively correlated with olfactory mucosal toxicity were identified as molecular dipolar momentum and the electronic properties of the substituent. Inversely correlated descriptors were variables describing the hydrophobicity, electronic properties of the molecule such as electron affinity and the electronic charge on the primary carbon. In conclusion, this preliminary PLS-DA model shows that a 2,6 dichlorinated benzene derivative with a large, polar, and strong electron withdrawing substituent in the primary position has the potential of being a potent OM toxicant in mice. PMID- 15536546 TI - Quantitative analysis of immobilized metalloenzymes by atomic absorption spectroscopy. AB - A new, sensitive assay for the quantitative determination of immobilized metal containing enzymes has been developed using atomic absorption spectroscopy (AAS). In contrast with conventionally used indirect methods the described quantitative AAS assay for metalloenzymes allows more exact analyses, because the carrier material with the enzyme is investigated directly. As an example, the validity and reliability of the method was examined by fixing the iron-containing enzyme catalase on cotton fabrics using different immobilization techniques. Sample preparation was carried out by dissolving the loaded fabrics in sulfuric acid before oxidising the residues with hydrogen peroxide. The iron concentrations were determined by flame atomic absorption spectrometry after calibration of the spectrometer with solutions of the free enzyme at different concentrations. PMID- 15536547 TI - A direct quantitative analysis method for monitoring biogenic volatile organic compounds released from leaves of Pelargonium hortorum in situ. AB - A direct quantitative method is presented that is based upon the use of multiple headspace solid phase microextraction (HS-SPME) to monitor biogenic volatile organic compounds (BVOCs) released from a living leaf of Pelargonium hortorum in situ. Seventeen BVOCs were detected by GC-MS after a single SPME extraction using a CAR/DVB/PDMS fibre. An internal standard was employed to determine the absolute amounts of seven terpenoid compounds released from a P. hortorum leaf. The quantitative analysis was performed over two days, with extraction preformed for 20 min every 3 h. The amount of volatiles extracted varied with the time of day, with two maxima recorded at 14:00 (day 1) and 17:00 (day 2), corresponding to 236 and 277 ng of the seven terpenoids recorded, respectively. These results indicate that multiple HS-SPME in combination with an internal standard is a simple, quick, and quantitative technique for analysising BVOC emissions from a live plant sample. PMID- 15536545 TI - Mapping dopamine D2/D3 receptor function using pharmacological magnetic resonance imaging. AB - RATIONALE: Regulation of dopamine release and synthesis occurs via pre-synaptic dopamine (DA) D2/D3 autoreceptors (DARs). Mapping of DAR function in vivo is difficult and is usually best assessed using invasive measures of DA release, such as microdialysis at discrete sites. We wished to show that pharmacological magnetic resonance imaging (phMRI) may prove useful for this purpose. OBJECTIVE: To demonstrate that the relative cerebral blood volume (rCBV) changes induced by amphetamine can be modulated by DA D2 receptor antagonists and agonists in a manner consistent with modulation of DAR function and to compare these effects with microdialysis. METHODS: We used phMRI with iron oxide contrast agents to map changes in rCBV in response to an amphetamine challenge, pre-treatment and post treatment with varying doses of the D2 antagonist eticlopride and the D2 agonist quinpirole. We also compared the effects of D2 antagonism using microdialysis measurements of DA release. RESULTS: Antagonism of D2 receptors with eticlopride potentiated rCBV changes induced by amphetamine in the nucleus accumbens and caudate putamen in a dose-dependent manner. The amphetamine-induced increase in rCBV in the accumbens in animals pre-treated with eticlopride was paralleled by a similar percentage increase in DA release measured by means of microdialysis. Conversely, agonism of D2 receptors using quinpirole reduced amphetamine-induced rCBV changes in the caudate putamen and nucleus accumbens. The effects of both quinpirole and eticlopride on amphetamine-induced rCBV changes were largest in the nucleus accumbens. CONCLUSIONS: These results suggest that phMRI may potentially prove useful to map DAR function non-invasively in multiple brain regions simultaneously. PMID- 15536548 TI - Enhanced signal generation for use in the analysis of synthetic pyrethroids using chemical ionization tandem quadrupole ion trap mass spectrometry. AB - Synthetic pyrethroids fragment extensively under electron ionization (EI) conditions to give low mass ions, most of them with the same m/z ratios. This fragmentation is primarily due to the labile ester linkage found in these compounds. In this research we established the best gas chromatography (GC) conditions in the EI mode that served as a benchmark in the development of a chemical ionization (CI) protocol for ten selected synthetic pyrethroids. Based on proton affinity data, several reagent gases were evaluated in the positive CI ionization mode. Methanol was found to produce higher average ion counts relative to the other gases evaluated, which led to the development of an optimized method consisting of selective ejection chemical ionization (SECI) and MS/MS. Standard stainless steel ion trap electrodes produced significant degradation of chromatographic performance on late eluting compounds, which was attributed to electrode surface chemistry. A dramatic improvement in signal-to-noise (S/N) ratios was observed when the chromatographically inert Silcosteel coated electrodes were used. The resulting method, that has significant S/N ratio improvements resulting from a combination of septum programmable injections (SPI), optimized CI and inert Silcosteel-coated electrodes, was used to determine instrument detection limits. PMID- 15536549 TI - Determination of thiol compounds in rat striatum microdialysate by HPLC with a nanosized CoHCF-modified electrode. AB - A cobalt hexacyanoferrate (CoHCF) nanoparticle (size ca. 60 nm) chemically modified electrode (CME) was fabricated and the electrochemical behavior of thiols at this nanosized CoHCF CME was studied. In comparison with a bare glassy carbon (GC) electrode and with a general CoHCF CME which was electrode-posited in the traditional manner, the present nanosized CoHCF CME efficiently performed electrocatalytic oxidation for glutathione (GSH) and L-Cysteine (L-Cys) with relatively high sensitivity, outstanding stability, and long-life. Combined with high-performance liquid chromatography (HPLC), the nanosized CoHCF CME was used for electrochemical determination (ECD) of GSH and L-Cys. The peak currents were a linear function of concentrations in the range 2.0 x 10(-7) to 2.0 x 10(-4) mol L(-1) for both GSH and L-Cys, with detection limits of 1.2 x 10(-7) and 1.0 x 10( 7) mol L(-1), respectively. Coupled with microdialysis sampling, the HPLC-ECD system has been successfully used to assess the GSH and L-Cys content of rat striatum. PMID- 15536550 TI - Characterisation of the interface of sputter-deposited copper coatings on nitrogen plasma-treated carbon substrates. AB - The adhesion of copper coatings to carbon substrates is very poor, because of lack of diffusion or reaction between the constituents. Because there is technological interest in enhancing the adhesion and improving the interface between copper and carbon, plasma treatment of the carbon substrate was employed in this study. For modification of the carbon surface a nitrogen plasma was used. It was confirmed by pull-off tests that the strength of adhesion of the copper coating can be improved by a factor of more than 10 by plasma pretreatment, even after treatment for a very short time (1 min). To obtain more information about the mechanisms of the processes occurring at the interface SIMS investigations were performed on samples which had been treated for different times (between 1 and 60 min). These measurements confirmed that nitrogen is located on the interface. With increasing pretreatment time the amount of nitrogen detected on the interface increased. Besides characterisation of as-deposited samples, another focus was to study the mechanisms of diffusion of nitrogen if the samples are heat treated at 500 degrees C. PMID- 15536551 TI - Consequences of increased neuromotor noise for reaching movements in persons with stroke. AB - Understanding stroke-induced changes to the motor control of the more affected arm of people with stroke may lead to more effective rehabilitation interventions that improve function. Reaching movements of the more affected arm in persons with stroke are slow, segmented, and indirect. Such changes may be related to a reduced capacity to transmit motor commands in the presence of neuromotor noise. In tasks requiring both speed and accuracy, transmission capacity can be characterized by the linear relationship between movement time and task difficulty (Fitts' law). This study quantified Fitts' slope and intercept coefficients in stroke during reaching tasks and their relationship to kinematic measures of path accuracy (directness), trajectory corrections (segmentation), and planning strategy (skewness). We compared Fitts' slope and intercept and kinematics among the more and the less affected arm of 20 persons with stroke and the nondominant arm of ten healthy persons. Slope and intercept were significantly increased in the more affected arm of the group with stroke and related to clinical measurements of motor impairment and tone. For both the more and the less affected arm of the group with stroke, increased slopes and intercepts were correlated to more indirect, segmented, and positively skewed movement. Our findings suggest that stroke results in greater neuromotor noise, which has consequences for both motor execution and planning. Individuals with stroke demonstrate substantially more deviation from straight-line paths than do controls, despite using more conservative strategies (i.e., leftward shift of velocity profile) and extensive feedback control (i.e., segmentation). PMID- 15536552 TI - Group II excitations from plantar foot muscles to human leg and thigh motoneurones. AB - Projections of group II afferents from intrinsic foot muscles to lower limb motoneurones were investigated in humans after electrical stimuli were applied to the tibial nerve (TN) at ankle level, using modulation of the quadriceps H reflex, on-going EMG of the quadriceps and peroneus brevis, and PSTHs of single quadriceps, biceps, semitendinosus, tibialis anterior, and peroneus brevis motor units. TN stimulation evoked late and high-threshold excitation in all leg and thigh muscles investigated. The mean latency of the late excitation was 13.5+/ 0.4 ms longer than that of the heteronymous monosynaptic Ia excitation, and the more caudal the motor nucleus the longer the central delay of the late effect, suggesting mediation through interneurones located rostral to motoneurones. The electrical threshold and conduction velocity of the largest diameter fibres evoking the late excitation were estimated to be approximately 2 and 0.67 times, respectively, those of the fastest Ia afferents, i.e. consistent with a mediation by group II afferents. Stimulation of the skin areas innervated by TN did not evoke late excitations. Further support for mediation through group II afferents was provided by the findings that: 1. the latency of the TN-induced late and high threshold excitation in Per brev units was more delayed by cooling the nerve than that of the excitation evoked by group I afferents, and 2. tizanidine intake (known to depress selectively transmission of group II effects) suppressed the TN induced late and high-threshold excitation whereas the group I facilitation was not modified. PMID- 15536553 TI - Discrete functional contributions of cerebral cortical foci in voluntary swallowing: a functional magnetic resonance imaging (fMRI) "Go, No-Go" study. AB - Brain-imaging studies have shown that visually-cued, voluntary swallowing activates a distributed network of cortical regions including the precentral and postcentral gyri, anterior cingulate cortex (ACC), insula, frontoparietal operculum, cuneus and precuneus. To elucidate the functional contributions of these discrete activation foci for swallowing, a "Go, No-Go" functional magnetic resonance imaging (fMRI) paradigm was designed. Brain activation associated with visually-cued swallowing was compared with brain activation evoked by a comparable visual cue instructing the subject not to swallow. Region-of-interest analyses performed on data from eight healthy subjects showed a significantly greater number of activated voxels within the precentral gyrus, postcentral gyrus, and ACC during the "Go" condition compared to the "No-Go" condition. This finding suggests that the precentral gyrus, postcentral gyrus, and ACC contribute primarily to the act of swallowing. In contrast, the numbers of activated voxels within the cuneus and precuneus were not significantly different for the "Go" and "No-Go" conditions, suggesting that these regions mediate processing of the cue to swallow. Together these findings support the view that the discrete cortical foci previously implicated in swallowing mediate functionally distinct components of the swallowing act. PMID- 15536554 TI - Dural enhancement with primary calvarial lesions. AB - The purpose of this study was to relate the pathological and imaging features of dural enhancement and meningeal sign ("dural tail") on contrast-enhanced T1 weighted magnetic resonance (MR) images from patients with primary calvarial lesions as well to assess the accuracy of MR imaging in predicting dural invasion. Thirty-two calvarial tumors studied with contrast-enhanced MR imaging and histopathological examination of the dural specimens were reviewed. Sixteen patients presented dural enhancement, eight with tumor invasion. Tumoral invasion of the dura was observed in one case without enhancement. Malignant lesions showed enhanced dura more commonly than benign lesions (P=0.02). Nodular and discontinuous dural enhancement was statistically associated with dural invasion (P=0.05). Dural tail did not show a specific pathological association. Meningeal enhancement is a nonspecific reaction to calvarial lesions unless nodular and discontinuous. False-negative and -positive cases of dural invasion imply some limitation of contrast-enhanced MR imaging in predicting dural invasion by calvarial neoplasms. PMID- 15536555 TI - Sensitivity and reproducibility of a new fast 3D segmentation technique for clinical MR-based brain volumetry in multiple sclerosis. AB - Fast, reliable and easy-to-use methods to quantify brain atrophy are of increasing importance in clinical studies on neuro-degenerative diseases. Here, ILAB 4, a new volumetry software that uses a fast semi-automated 3D segmentation of thin-slice T1-weighted 3D MR images based on a modified watershed transform and an automatic histogram analysis was evaluated. It provides the cerebral volumes: whole brain, white matter, gray matter and intracranial cavity. Inter- and intra-rater reliability and scan-rescan reproducibility were excellent in measuring whole brain volumes (coefficients of variation below 0.5%) of volunteers and patients. However, gray and white matter volumes were more susceptible to image quality. High accuracy of the absolute volume results (+/-5 ml) were shown by phantom and preparation measurements. Analysis times were 6 min for processing of 128 slices. The proposed technique is reliable and highly suitable for quantitative studies of brain atrophy, e.g., in multiple sclerosis. PMID- 15536556 TI - Morphologic changes in neo-intimal proliferation in an experimental aneurysm after coil embolization: effect of factor XIII administration. AB - We investigated histological changes in aneurysmal orifices after embolization with Guglielmi detachable coils (GDCs) and determined the effects of the wound healing factor, factor XIII, on promoting intimal proliferation by scanning electron microscopy (SEM). GDC embolization was performed in an experimental model of aneurysm in swine. In the control group (17 aneurysms), the aneurysms were resected immediately after surgery, at 1 and 3 weeks after the procedure. In the factor XIII-administered group (13 aneurysms), the swine received factor XIII postoperatively, and the aneurysms were excised at 1 and 3 weeks. The endothelial cell proliferation changes in the aneurysm orifices in both groups were evaluated by SEM. The histological changes at the orifices began immediately after the procedure, and endothelialization was observed at 1 week. One week after the procedure, the rate of endothelial cell proliferation was significantly higher in the factor XIII group (P<0.05). But no difference was observed at 3 weeks, when endothelialization of the orifices was essentially completed. The process of intimal proliferation after coil embolization was similar to the wound-healing process after vascular intimal injury. Administration of the wound-healing factor, factor XIII, would contribute rapid intimal proliferation and may be effective to facilitate complete obliteration of aneurysms after coil embolization. PMID- 15536557 TI - Atypical manifestations of reversible posterior leukoencephalopathy syndrome: findings on diffusion imaging and ADC mapping. AB - Typically, reversible posterior leukoencephalopathy syndrome (RPLS) involves the parieto-occipital lobes. When regions of the brain other than the parieto occipital lobes are predominantly involved, the syndrome can be called atypical RPLS. The purpose of this study is to find radiological and pathophysiological features of atypical RPLS by using diffusion-weighted imaging (D-WI). We retrospectively reviewed seven patients (two with eclampsia, one with cyclosporine neurotoxicity, and four with hypertensive encephalopathy) with atypical MR manifestations of RPLS. Changes in signal intensity on T2-weighted imaging (T2-WI) and D-WI, and ADC ratio, were analyzed. In patients with atypical manifestation of RPLS, high signal intensities on T2-WI were noted in the frontal lobe, basal ganglia, thalamus, brainstem, and subcortical white matter in regions other than the parieto-occipital lobes. These areas of increased signal intensities on T2-WI showed increased ADC values, representing vasogenic edema in all seven patients. This result should be very useful in differentiating atypical RPLS from other metabolic brain disorders that affect the same sites with cytotoxic edema. PMID- 15536559 TI - Transvenous injection of Onyx for casting of the cavernous sinus for the treatment of a carotid-cavernous fistula. AB - A complex case of carotid-cavernous fistula was treated transvenously by injection of ethyl vinyl alcohol co-polymer into the cavernous sinus after an unsuccessful embolization attempt with detachable coils and liquid adhesive agents. There were no complications. At 3 months the patient's symptoms had resolved completely, and a control angiogram revealed persistent occlusion. The physical properties of ethyl vinyl alcohol polymer justify further investigation of this agent for the treatment of carotid-cavernous fistula. PMID- 15536560 TI - American Board of Radiology update. PMID- 15536561 TI - Malrotation with midgut volvulus: CT findings of bowel infarction. AB - Midgut volvulus, the most common serious complication of malrotation, can be diagnosed using conventional contrast fluoroscopy, US or CT. CT is a quick and comprehensive examination in the evaluation of complex acute abdominal pathology in children. Contrast-enhanced CT can readily help the radiologist recognize perfusion abnormalities of the bowel, which is vital for reducing morbidity and mortality in affected children. Our case emphasizes and demonstrates additional CT features of bowel infarction in a child with a proven malrotation with midgut volvulus. PMID- 15536562 TI - Disorders of the pediatric pancreas: imaging features. AB - The purpose of this manuscript is to provide an overview of the normal development of the pancreas as well as pancreatic pathology in children. Diagnostic imaging plays a major role in the evaluation of the pancreas in infants and children. Familiarity with the range of normal appearance and the diseases that commonly affect this gland is important for the accurate and timely diagnosis of pancreatic disorders in the pediatric population. Normal embryology is discussed, as are the most common congenital anomalies that occur as a result of aberrant development during embryology. These include pancreas divisum, annular pancreas, agenesis of the dorsal pancreatic anlagen and ectopic pancreatic tissue. Syndromes that can manifest pancreatic pathology include: Beckwith Wiedemann syndrome, von Hippel-Lindau disease and autosomal dominant polycystic kidney disease. Children and adults with cystic fibrosis and Shwachman Diamond syndrome frequently present with pancreatic insufficiency. Trauma is the most common cause of pancreatitis in children. In younger children, unexplained pancreatic injury must always alert the radiologist to potential child abuse. Pancreatic pseudocysts are a complication of trauma, but can also be seen in the setting of acute or chronic pancreatitis from other causes. Primary pancreatic neoplasms are rare in children and are divided into exocrine tumors such as pancreatoblastoma and adenocarcinoma and into endocrine or islet cell tumors. Islet cell tumors are classified as functioning (insulinoma, gastrinoma, VIPoma and glucagonoma) and nonfunctioning tumors. Solid-cystic papillary tumor is probably the most common pancreatic tumor in Asian children. Although quite rare, secondary tumors of the pancreas can be associated with certain primary malignancies. PMID- 15536563 TI - L-type calcium channel gating is modulated by bradykinin with a PKC-dependent mechanism in NG108-15 cells. AB - Bradykinin (BK) excites dorsal root ganglion cells, leading to the sensation of pain. The actions of BK are thought to be mediated by heterotrimeric G protein regulated pathways. Indeed there is strong evidence that in different cell types BK is involved in phosphoinositide breakdown following activation of G(q/11). In the present study we show that the Ca(2+) current flowing through L-type voltage gated Ca(2+) channels in NG108-15 cells (differentiated in vitro to acquire a neuronal phenotype), measured using the whole-cell patch clamp configuration, is reversibly inhibited by BK in a voltage-independent fashion, suggesting a cascade process where a second messenger system is involved. This inhibitory action of BK is mimicked by the application of 1,2-oleoyl-acetyl glycerol (OAG), an analog of diacylglycerol that activates PKC. Interestingly, OAG occluded the effects of BK and both effects were blocked by selective PKC inhibitors. The down modulation of single L-type Ca(2+) channels by BK and OAG was also investigated in cell attached patches. Our results indicate that the inhibitory action of BK involves activation of PKC and mainly shows up in a significant reduction of the probability of channel opening, caused by an increase and clustering of null sweeps in response to BK. PMID- 15536564 TI - Studying the effect of a charged surface on the interaction of bleomycin with DNA using an atomic force microscope. AB - The cleavage of DNA caused by the antitumoral drug bleomycin has been investigated using atomic force microscopy (AFM). This work deals with the effect that adsorbing DNA onto a positively- or negatively-charged surface has on the double-strand cleavage of DNA by Fe(III)/bleomycin. Quantitative analysis of the number of breaks per DNA molecule, in bulk and at the surface of the mica substrate, has been performed by analyzing AFM images. It turns out that the cleavage of DNA is strongly inhibited by a positively-charged surface. Our experiments can be interpreted using a simple electrostatic model. This paper is a first step in the study of DNA accessibility to ligand such as bleomycin, using AFM in liquids. PMID- 15536565 TI - Influence of protein flexibility on the electrostatic energy landscape in gramicidin A. AB - We describe an electrostatic model of the gramicidin A channel that allows protein atoms to move in response to the presence of a permeating ion. To do this, molecular dynamics simulations are carried out with a permeating ion at various positions within the channel. Then an ensemble of atomic coordinates taken from the simulations are used to construct energy profiles using macroscopic electrostatic calculations. The energy profiles constructed are compared to experimentally-determined conductance data by inserting them into Brownian dynamics simulations. We find that the energy landscape seen by a permeating ion changes significantly when we allow the protein atoms to move rather than using a rigid protein structure. However, the model developed cannot satisfactorily reproduce all of the experimental data. Thus, even when protein atoms are allowed to move, the dielectric model used in our electrostatic calculations breaks down when modeling the gramicidin channel. PMID- 15536566 TI - Polymer-cushioned lipid bilayers in porous alumina. AB - Using small-angle neutron scattering (SANS) and cyclic voltammetry (CV), we show that model biological membranes can be deposited on a polymer cushion confined in highly regular porous alumina. The thicknesses of the dilute polymer cushion chemically bound to the alumina and of the supported bilayer are obtained for two polyethylene glycol cushions (PEG(5000) and PEG(20000)) and for a cushion made of chains bearing a lipid anchor at their free end (DSPE-PEG(3400)). The bilayers are studied well below and well above the chain melting temperature of the lipid mixture (DMPC/DMPE: 80/20), using a coenzyme (Ubiquinone, UQ(10)) as a redox probe for the voltammetry experiments. Analysis of the SANS form factor of the bilayers shows that the bilayer thickness can be extracted in this particular geometry. Using PEG chains grafted at a low surface density (D < 2R(g)), the thickness of the complete molecular construction is obtained by CV, which shows (after subtracting the bilayer thickness) that the polymer cushion thickness can be varied from 50 to 150 Angstroms. The values obtained with three different chain lengths, are in perfect agreement with the radius derived from the Flory theory. PMID- 15536568 TI - [Pain treatment in burn patients]. AB - Burn injuries can be causing the most intense and prolonged types of pain. Pain in such patients can also cause psychologic and functional difficulties, and is difficult to predict from wound depth. The fundamental problem of the burn pain treatment is insufficient analgesia. The pain management of the burn patient can be very difficult and necessitate enough experience because of individual differences of the patients and pain that involve different components. The psychological support and treatment is also as important as pharmacological treatment. The success rate of the pain treatment of the burn patient can be increased with the multidisciplinary approach based on the decision of the most appropriate pain treatment modalities for individual patient and the principles of the pain treatment. PMID- 15536567 TI - Detection of a new cerebral malaria susceptibility locus, using CBA mice. AB - Human cerebral malaria (CM) during acute Plasmodium falciparum infection is a serious neurological complication that leads to coma and death. P. berghei ANKA infection of CBA mice is a useful experimental model of CM. To identify host susceptibility loci, we performed chromosomal mapping in crossbred populations of both CM-susceptible CBA and CM-resistant DBA/2 mice. One significant region for a CM-susceptible locus in CBA mice was mapped to H2 region on Chromosome 17, tentatively designated cmsc. cmsc was mapped to a different chromosomal region from that previously reported in the C57BL/6 mouse model of CM. It is possible that different loci contribute to CM in CBA and C57BL/6 mouse strains. Comparison of the function of CM susceptibility loci between CBA and C57BL/6 mice could have important implications for the study of the complex pathogenesis of CM in humans. PMID- 15536569 TI - [Trigeminal autonomic cephalgias: diagnosis, therapy, atypical forms and pathophysiology]. AB - Trigeminal autonomic cephalgias; cluster headache, paroxysmal hemicrania, hemicrania continua and SUNCT (short-lasting unilateral neuralgiform headache attacks with conjuctival injection and tearing) syndrome are characterized by unilateral trigeminal distribution of pain and accompanying ipsilateral autonomic symptoms. Other than cluster headache, all of these syndromes have been described within last twenty years, and are found relatively less frequent and less known forms. Diagnosis of paroxysmal hemicrania and hemicrania continua, especially atypical forms, essentially depends on indomethacin responsiveness. For SUNCT syndrome, there is not such a drug which provides a practical approach to both diagnosis and therapy and its diagnosis depends on clinical features. So far, case reports from various countries helped us realize the existence of secondary forms of these syndromes and the necessity of imaging techniques, especially for recently described autonomic cephalgias. PMID- 15536570 TI - Hypnic headache syndrome: excessive periodic limb movements in polysomnography. AB - We report a patient with hypnic headache syndrome associated with excessive periodic limb movements in sleep, which is a unique finding for this syndrome recorded in polysomnography. She had had daily hypnic headache attacks history for 10 years. Her headache attacks ceased immediately after lithium carbonate therapy and she has been headache-free for 5 months. PMID- 15536571 TI - [Cancer pain management in children]. AB - Pain is one of the most common and frightening symptoms of children with cancer and their families. If current pain management techniques are utilized, the majority of children can achieve adequate analgesia. It is the rare pediatric patients who develop intractable pain. The World Health Organization (WHO) has established principles of pain management for children with cancer. PMID- 15536572 TI - Intrathecal melatonin increases the mechanical nociceptive threshold in the rat. AB - The aim of this investigation was to determine whether intrathecal (i.t.) administration of monodose melatonin provides an increase in mechanical nociceptive threshold in the rat. Twenty-four male Wistar rats were randomly assigned to four groups. Each animal was anesthetized, and a catheter was placed intrathecally via the cisterna magna. The study groups were: untreated controls (n=6); melatonin only (MEL, n=6); melatonin + luzindole (MEL+LZN, n=6); and melatonin + naloxone (MEL+NLX, n=6). Measurements of mechanical nociceptive threshold were made using an electronic algometer. Each animal was tested prior to injection and at 10, 20, 30 and 40 min after injection. In the MEL group, the mean nociceptive thresholds at all post-injection time points were significantly higher than the baseline value (p<0.05 for all). In the control and MEL+LZN groups, none of the four mean nociceptive thresholds recorded after i.t. injection was significantly different from the baseline value (p>0.05 for all). In the MEL+NLX group, the mean nociceptive thresholds at 20, 30 and 40 min post injection were all significantly lower than the baseline value (p<0.05 for all). Comparison among the group nociceptive thresholds at baseline revealed no significant differences, and the same was true at 10 min after i.t. injection. At the 20, 30 and 40 min stages, the threshold in the MEL group was significantly higher than the threshold in the control group. The results indicate that i.t. injection of melatonin produces a time-dependent increase in mechanical nociceptive threshold in the rat and that the mechanism that underlies these effects involves both melatonin and opioid receptors. PMID- 15536573 TI - Percutaneous annuloplasty in the treatment of discogenic pain: retrospective evaluation of one year follow-up. AB - In this study, we have evaluated clinical results of the discTRODE system, in 15 patients with discogenic pain. All procedures were performed under sterile conditions with fluoroscopic guidance. After identifying the disc space under fluoroscopy, the introducer of the discTRODE was introduced using a tunneled vision. After inserting the introducer; navigable, semi-rigid discTRODE catheter was advanced through the introducer and directed medially and contralaterally along the posterior nuclear-annular interface. The SMK Thermocouple Electrode was placed in the outer-annulus on the contralateral side so as to monitor local tissue temperature. The treatment temperature was manually increased in a step wise progression from 50 degrees C to 65 degrees C. Patient outcomes were evaluated during follow-up visits at 1, 3, 6 and 12th months post-procedure. Before the procedure and at each visit during the follow-up period, patients completed Visual Analogue Pain Scale (VAS) and Short Form-36 Health Status Questionnaire (SF-36). Annuloplasty was performed without difficulty in all patients, there were no complications associated with disc puncture such as discitis or disc rupture. Symptoms improved in 10 of 15 (66.6 %) of the patients on the SF-36 Physical Function subscale, in 9 of 15 (60 %) on the SF-36 Bodily Pain subscale and in 9 of 15 (60 %) on the VAS scores. 5 of 15 of the patients did not show improvement on any scale. This technique seems to be a reliable method for patients complaining of discogenic pain. However, prospective randomized controlled studies comparing different approaches are needed. PMID- 15536574 TI - [Blockade of ganglion impar through sacrococcygeal junction for cancer-related pelvic pain]. AB - Impar ganglion block provides pain relief in patients who suffer from sympathetically mediated pain arising from disorders of viscera and somatic structures within the pelvis and perineum. We performed impar ganglion blockade through sacrococcygeal junction instead of anococcygeal ligament in 9 patients who had localized perineal pain of visceral origin. All the blocks which were performed through sacrococcygeal junction without bending the needle under fluoroscopic guidance were easy to perform without any complication. Pain intensity by Visual Analogue Scale, daily opioid requirements, and complications related to opioids were evaluated before the procedure, and for 2 months after the procedure. The intensity of pain, daily opioid requirement and the complication related to the opioids were significantly decreased in 8 patients. We suggest that impar ganglion block through sacrococcygeal junction appears to be safe and effective procedure in the management of perineal pain related to malignancy. PMID- 15536575 TI - [Beneficial effects of single dose multimodal epidural analgesia on relief of postoperative microdiscectomy pain]. AB - We aimed to assess the efficacy of multimodal epidural analgesia in decreasing postoperative pain after microdiscectomy. Fourty patients, ASA physical status I or II, undergoing microsurgical lumbar discectomy were enrolled in this prospective, randomised, controlled, double-blinded study. 10 ml study solution consisting of 2 mg of morphine, 15 mg of bupivacaine, 80 mg of methylprednisolone, and 0.05 mg of adrenaline was prepared for epidural administration. At the end of the procedure but prior to wound closure, the surgeon inserted an 18-gauge epidural catheter into the epidural space. After closure of incision, patients were assigned to receive either study solution (Group E) or saline (Group C). The epidural catheter was then removed. Patient controlled analgesia with morphine was used for postoperative analgesia. Visual Anologue Scale (VAS) pain scores and morphine consumptions were lower in Group E. Time to first ambulation was shorter in Group E. Patients in Group E were more satisfied with their analgesic regimen. Single dose multimodal epidural analgesia administered after wound closure provided better postoperative analgesia after lumbar microdiscectomy. PMID- 15536576 TI - Comment on: Isin Unal Cevik: Postherpetic neuralgia. Agri 2004; 16(3): 17-24. PMID- 15536578 TI - [Advances in hypertension research]. PMID- 15536579 TI - [Information deficits concerning blood pressure self-measurement]. AB - BACKGROUND AND OBJECTIVE: Blood pressure values resulting from measurements by the patients themselves have greater diagnostic and prognostic significance than those measured by the physician, assuming, however, that measurement is taken correctly. As self-measurement of blood pressure has meanwhile become a widespread practice. The question arises to what extent the rules on exact measurement of blood pressure are known to the patient. PATIENTS AND METHODS: The answers to questions on five rules on self-measurement of blood pressure (resting period prior to measurement, time of measurement and intake of morning medication, left or right arm for measurement, level of measuring point and width of cuff) given by 500 hypertensive patients (373 men, 127 women, mean age 59.2 +/ 10.2 years) were evaluated. RESULTS: Three patients only were in the position to answer all five questions correctly. 19 % were unable to give any right answer. Patients showed highest information levels concerning the resting period before measurement. Approximately half of them (51 %) knew that a pause of three minutes has to be kept. Only one in three, however, was familiar with the fact that morning measurement has to be taken prior to the intake of antihypertensives (32 %) choosing the arm that shows higher blood pressure values. 28 % were informed that the measuring point has to lie at the height of the heart. Only 4 % were able to answer the question on the width of the cuff correctly. CONCLUSION: Our findings show that the rules on correct self-measurement of blood pressure are unknown to most of the hypertensive patients. This may be due to the fact that they are insufficiently informed and trained. PMID- 15536580 TI - [Recurrent hemispheric symptoms without proof of ischemia]. AB - ANAMNESIS AND CLINICAL FINDINGS: Over the last 6 years, a 48-year-old patient recurrently experienced passing neurological left hemispheric symptoms of varying strength. Because of the patient's distinctive cardiovascular risk profile, these symptoms were diagnosed as transitory ischaemic episodes. On inquiry, the patient now reported frequent headaches in close temporal connection to the events. EXAMINATIONS: Repeated imaging of the neocranium using various techniques (CCT, cMRT, MRT-angiography) revealed no conspicuous features. Duplex sonography of the brain feeding arteries, echocardiography, EEG and 24h-ECG also gave findings within the normal range for the patient's age group. Clinical chemistry showed increased values for cholesterol and triglycerides. Incidentally, a moderate upper airway resistance syndrome was identified during polysomnography. DIAGNOSIS: Suspected migraine with aura in terms of a sporadic hemiplegic migraine. TREATMENT AND COURSE: On administration of ASS and rizatriptane, the neurological hemispheric symptoms disappeared completely over the course of a few hours. Under prophylactic treatment with metoprolol and acetazolamide over two months, no further events have been observed up to now. CONCLUSION: Recurrent, temporary neurological deficits, together with the presence of several cardiovascular risk factors, can easily be misinterpreted to be the result of an underlying cerebral ischemia. Even in cases with typical cardiovascular risk profile, a migraine with aura must be considered as one of the possible causes for hemispheric neurological symptoms. PMID- 15536581 TI - [Cardiovascular risk and renal failure]. PMID- 15536582 TI - [24-hour blood pressure and ECG monitoring in patients with arterial hypertonia - importance of silent myocardial ischemia]. PMID- 15536583 TI - [The importance of systemic hypertension for eye diseases]. PMID- 15536584 TI - [Old and new drugs to block aldosterone and ameliorate cardiovascular disease]. PMID- 15536585 TI - [Role of stem- and progenitor cells in coronary artery disease]. PMID- 15536586 TI - Factors associated with American Indian teens' self-rated health. AB - Factors related to American Indian (AI) high school students' self-rated health were examined. Self-rated health was measured as a single-item with a four-point response option ranging from poor to excellent health. Of the 574 participants, 19% reported "fair" or "poor" health, a percentage more than twice that for U.S. high school students in general. Gender (related to family finances), school achievement, social competence, and cannabis use were significantly associated with the AI teens' self-rated health. In comparative examination, factors associated with AI teens' health ratings were found to be similar and dissimilar to ratings of other teens in important ways. PMID- 15536587 TI - Comparing three measures of depressive symptoms among American Indian adolescents. AB - This study examined the measurement of depressive symptoms among American Indian adolescents as assessed by the Center for Epidemiologic Studies Depression Scale (CES-D), Youth Self Report (YSR), and the Tri-Ethnic Center's for Prevention Research Depression Scale (TEDS). This analysis demonstrated that the TEDS had good internal consistency, demonstrated construct validity, and shared a commonality with the other two measures, but had limited predictive validity. The YSR had strong predictive validity but, like the CES-D, showed weakness in construct validity. Unexpectedly, the CES-D and YSR revealed enculturated youth were at risk of depressive symptoms. These findings do not generate unequivocal support for any one measure, but suggest that their strengths and potential shortcomings should be taken into account when assessing depressive symptoms among American Indian youth. PMID- 15536588 TI - Antidepressant medication use among First Nations peoples residing within British Columbia. AB - Very little is known about antidepressant medication use among First Nations people in Canada. This information would be useful to begin estimating the prevalence of conditions treated with this class of medications and planning appropriate programs. Antidepressant medication claims for First Nations people residing within British Columbia were extracted from the Non-Insured Health Benefits pharmacy database. During 2001, 9.8% (95% CI = 9.81, 9.79) of the population filled a prescription for antidepressant pharmacotherapy, claimant mean age was 40.3 years and the female:male ratio was approximately 3:1. The most common medications were Paxil, Apo-Amitriptyline, Effexor, and Celexa. Use of this medication class is common and more research is needed in this area of study. PMID- 15536589 TI - Intimate partner violence in American Indian and/or Alaska Native communities: a social ecological framework of determinants and interventions. AB - This essay synthesizes the research on intimate partner violence (IPV) in American Indian and/or Alaska Native communities using a social ecological framework. The review of literature demonstrates that American Indian and/or Alaska Native women are at an elevated risk for IPV compared to non-American Indian women and thus this essay describes multi-level interventions that are culturally appropriate for American Indian and/or Alaska Native communities. The interventions address a variety of determinants including gender, age, socioeconomic status, alcohol, European colonization, and infrastructure. PMID- 15536590 TI - Cardiac dysfunction in the euglycemic diabetic-prone BB Wor rat. AB - The purpose of this study was to examine cardiac function in the diabetic-prone BB Wor rat. The study involved 2 groups: diabetic resistant control littermates of BB rats and diabetic-prone BB rats that had yet to demonstrate overt signs of diabetes. Hearts from these animals were isolated and cardiac function examined in response to incremental increases in left atrial filling pressure. Hearts were also perfused at an increased aortic afterload resistance with buffer consisting of glucose alone or glucose in the presence of palmitate. Hearts from diabetic prone rats exhibited depressed contractility and ventricular relaxation at high filling pressures. Ventricular function, expressed as cardiac output, was also depressed in diabetic-prone rats perfused at increased afterload resistance, but only in the presence of palmitate. Our results indicate that hearts from diabetic prone BB Wor rats demonstrate abnormalities in contractile performance and thus may be a useful model for the study of cardiac function in the prediabetic state. PMID- 15536591 TI - Increased resting metabolic rate in patients with type 2 diabetes mellitus accompanied by advanced diabetic nephropathy. AB - Thirty-three patients with type 2 diabetes mellitus (16 men, 17 women) were divided into 3 groups based on urinary excretion of albumin (U-Alb)--group A: U Alb < 30 mg/d; group B: 30 mg/d < or = U-Alb < or = 300 mg/d; and group C: 300 mg/d < U-Alb. Serum creatinine levels were lower than 2.0 mg/dL in all the subjects. There was no difference in age, sex, therapy, body weight, body mass index (BMI), lean body mass (LBM), or hemoglobin A(1c) (HbA(1c)) levels among the 3 groups. Resting metabolic rate (RMR) (kJ/h/m(2)) and adjusted RMR for lean body mass (kJ/h/m(2)) were significantly increased in group C compared with groups A and B. Hb concentrations, serum albumin levels, and creatinine clearance were much lower in group C than in groups A and B (P < .001). There were no difference in serum urea nitrogen, total cholesterol, cholinesterase and free thyroxine, or plasma insulin-like growth factor I (IGF-I) levels among the 3 groups. Linear regression analysis revealed an inverse correlation between RMR and serum albumin levels, correlation between RMR and U-Alb, and inverse correlation between RMR and Hb concentrations, respectively, in these patients. In conclusion, RMR in diabetic patients correlated directly with U-Alb and inversely with serum albumin and Hb concentration. These findings suggest that RMR is related with urinary albumin loss and anemia in patients with type 2 diabetes mellitus accompanied by diabetic nephropathy. PMID- 15536592 TI - Stability over time of glycohemoglobin, glucose, and red blood cell survival in hematologically stable people without diabetes. AB - We previously have shown that an affinity, high-performance liquid chromatography (HPLC) method is a highly reproducible and sensitive method for determining percent total glycohemoglobin (tGHb) in people with diabetes. In this study we extended the use of this method to a determination of the correlation of percent tGHb with the fasting plasma glucose concentration in people without known diabetes. We also determined the correlation of the tGHb with the reticulocyte count, as an index of red blood cell (RBC) survival, and with a carbon monoxide (CO) method for determining RBC survival. In addition, the stability of the tGHb, glucose, RBC mass, hemoglobin, and reticulocyte counts over a 1-year period was evaluated. Total glycohemoglobin, overnight fasting plasma glucose concentration, hemoglobin, RBC and reticulocyte count, and the calculated percentage of RBC count represented by reticulocytes were determined monthly for at least 12 months (range, 12 to 26 months) in 48 adults (mean age, 51 years; range, 31 to 82 years). In 37 of the subjects, RBC survival using a CO method also was determined. There was a highly significant linear correlation between the fasting glucose concentration and the tGHb. There was only a weak correlation between the percent reticulocytes or with the RBC survival determined by the CO method. The tGHb, plasma glucose, RBC count, hemoglobin, and percent reticulocytes were very stable over a 12-month or greater period. We conclude that there is a good correlation between the tGHb and plasma glucose concentration in a population without known diabetes. Variations in RBC survival as indicated by a reticulocyte count within the reference range is not likely to have a clinically significant effect on interpretation of tGHb data in the context of an integrated glucose concentration. Nevertheless, this remains to be proven using RBC survival methods that are more precise than those currently available. PMID- 15536593 TI - Cilostazol, a phosphodiesterase inhibitor, reduces microalbuminuria in the insulin-resistant Otsuka Long-Evans Tokushima Fatty rat. AB - We evaluated association between hyperinsulinemia/insulin resistance and microalbuminuria in the insulin-resistant Otsuka Long-Evans Tokushima Fatty (OLETF) rat. OLETF rats showed glomerular hyperfiltration (an increase in creatinine clearance and a decrease in fractional excretion of Na) and microalbuminuria at the insulin-resistant prediabetic stage, and both were related to expression of transforming growth factor (TGF)-beta(1) and extracellular matrix protein such as fibronectin and collagen (a(1)) IV. Cilostazol, a selective type III cyclic nucleotide phosphodiesterase (PDE) inhibitor, normalized glomerular hyperfiltration and microalbuminuria with a parallel decline of TGF-beta(1) and extracellular matrix protein mRNA expression. Cilostazol may be beneficial to lessen early glomerular nephropathy in a state of hyperinsulinemia/insulin resistance. PMID- 15536595 TI - Adaptive thermogenesis is intact in B6 and A/J mice studied at thermoneutrality. AB - To investigate mechanisms of resistance to obesity, the physiologic responses to short-term moderate fat feeding were studied at ambient temperature (T(a)) = 23 degrees C and thermonuetrality (T(a) = 30 degrees C) in mice susceptible (B6) or resistant (A/J) to obesity. We hypothesized that A/J mice would exhibit greater adaptive thermogenic responses to consumption of moderate-fat diets, and that this response would be attenuated in thermoneutral conditions due to reduced activity of brown adipose tissue (BAT). B6 and A/J mice were adapted to either T(a) = 23 degrees C or T(a) = 30 degrees C, implanted with telemetry devices, housed in metabolic chambers for measurement of food intake, oxygen consumption (Vo(2)), and heart rate (HR), and studied before and during 1 week of consuming a diet containing 32% of calories from fat. Access to 32% fat diet resulted in increased caloric intake in both strains, but caloric intake for A/J mice returned to baseline levels within 72 hours, while B6 mice remained hyperphagic. Both strains exhibited increased light-phase Vo(2) indicative of adaptive thermogenesis; however, there was no strain difference in light-phase Vo(2) during the 1-week feeding trial. Surprisingly, T(a) had no effect on diet-induced thermogenesis in either mouse strain. Moderate high-fat feeding produced mild tachycardia that was similar in B6 and A/J mice and more clearly evident at thermonuetrality. We conclude that adaptive thermogenic responses are intact in both mouse strains studied at thermoneutrality, suggesting a minimal role for BAT in the initial metabolic response to hyperphagia. Furthermore, the results suggest that differences in control of caloric intake, rather than capacity for adaptive thermogenesis, may contribute to the relative susceptibility to obesity in A/J and B6 mice. PMID- 15536594 TI - The Arg64 allele of the beta 3-adrenoceptor gene but not the -3826G allele of the uncoupling protein 1 gene is associated with increased leptin levels in the Spanish population. AB - To determine whether there are variations in leptin levels according to the beta(3)-adrenoceptor (beta(3)-AR) Trp64Arg and uncoupling protein 1 (UCP1) -3826A ->G polymorphisms, given the regulatory role of catecholamines through the beta(3)-AR in leptin production and the previously reported association of the UCP1 -3826A-->G variant with obesity. A total of 160 men and 172 women randomly chosen from a nationwide population-based obesity cross-sectional survey in Spain were studied. Body mass index (BMI), waist-to-hip ratio (WHR), leptin, insulin, fasting and 2-hour post-glucose load glycemia, high-density lipoprotein (HDL)-, low-density lipoprotein (LDL)-, and total cholesterol, and triglyceride plasma levels were measured. beta(3)-AR Trp64Arg and UCP1 -3826A-->G genotypes were determined by restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR). UCP1 -3826G allele frequency was higher in men than in women (0.31 v 0.22, P = .015) and in obese women than in non-obese women (0.31 v 0.17, P = .008). Women carriers of the Arg64 or the alleles also showed higher leptin levels than noncarriers. Multiple linear regression analysis showed that the Arg64 allele is associated with higher leptin levels after the adjustment for gender, age, WHR, and the degree of glucose tolerance. In conclusion, the beta(3) AR Trp64Arg polymorphism might have an impact on the mechanisms involved in leptin release from adipose tissue. Furthermore, our results agree with the previously reported association between UCP1 -3826G allele and obesity and point to a gender-related effect. PMID- 15536596 TI - Desensitization of the inhibitory effect of norepinephrine on insulin secretion from pancreatic islets of exercise-trained rats. AB - The effect of exercise training (9 weeks of running) on norepinephrine-induced inhibition of insulin secretion was examined in rat islets. Insulin secretions from islets in the presence of glucose (> or =5.5 mmol/L) were significantly lower in trained (TR) than in control rats (CR). Norepinephrine inhibited 5.5 mmol/L glucose-stimulated insulin secretions and cyclic adenosine monophosphate (cAMP) contents in a dose-dependent manner in CR. Norepinephrine (10 micromol/L) induced inhibition of insulin secretion was reversed by the blockade of the alpha(2)-adrenergic receptor in CR, but not in TR. Exercise training substantially shifted the dose-dependent curve for clonidine-induced inhibition of insulin secretions and that of cAMP contents to the right. Exercise training did not alter the density of the alpha(2)-adrenergic receptor either per islet or per protein of islet crude membrane. However, exercise training significantly reduced the protein expression of G alpha i-2 without change in G alpha i-2 mRNA. In CR but not in TR, norepinephrine significantly inhibited insulin secretions elicited by a combination of high glucose, a protein kinase C activator, and an adenylate cyclase activator under Ca(2+)-free conditions. Thus, exercise training appears to provoke a decreased expression of G alpha i-2 protein. This, at least in part, results in loss of the inhibitory effect of norepinephrine either on cAMP content or on insulin secretion at the post-calcium events in stimulus secretion coupling, which, in turn, leads to the blunted inhibitory effects of norepinephrine on insulin secretion. PMID- 15536598 TI - Association between diet, lifestyle, metabolic cardiovascular risk factors, and plasma C-reactive protein levels. AB - Increased C-reactive protein (CRP) levels have been associated with several of the components of the metabolic syndrome, but the direct influence of diet and lifestyle factors on CRP levels remains largely unknown. The purpose of the present study was to investigate the association between CRP and diet and lifestyle factors. Plasma CRP levels were determined by a highly sensitive enzyme linked immunosorbent assay (ELISA) in 760 participants in the beta-Blocker Cholesterol-Lowering Asymptomatic Plaque Study (BCAPS). In accordance with previous findings, increased levels of CRP were associated with high body mass index (BMI) (P = .012), triglycerides (P = .001), systolic blood pressure (P = .019), cholesterol/high-density lipoprotein (HDL) ratio (P = .009), and low HDL cholesterol (P = .001). CRP was also increased in smokers (P = .023) and in subjects with a low vitamin C intake (P = .018). When men and women were analyzed together, there were no significant associations between CRP and dietary intake of total calories, total fat, saturated fat, monounsaturated fat, polyunsaturated fat, n-3 polyunsaturated fatty acids, n-6 polyunsaturated fatty acids, fiber, vitamin E, carotene, or selen, or in physical activity. However, in the female subgroup weak inverse relations were observed between CRP and the intake of total fat (r = -0.13, P = .011), saturated fat (r = -0.13, P = .011), monounsaturated fat (r = -0.13, P = .010), polyunsaturated fat (r = -0.14, P = .007), and n-3 PUFA (r = -0.14, P = .004). Stratified factor analyses in smoking subgroups, obese, and in under-reporters of energy, largely confirmed the results although in male never-smokers a combination of high fiber vitamin C/beta carotene intake was associated with low CRP levels. These observations suggest that CRP levels are only marginally associated with individual dietary and lifestyle factors. Surprisingly, a higher intake of fat tended to be associated with lower CRP values among women. PMID- 15536597 TI - Is there any link between insulin resistance and inflammation in established preeclampsia? AB - Both insulin resistance and inflammation may contribute to the onset of preeclampsia. They also could be interrelated. We studied the relationship between inflammatory cytokines and markers of insulin resistance. During their third trimester, 22 proteinuric preeclamptic women and 16 normotensive controls underwent intravenous glucose tolerance test (minimal model). Preeclamptic women were more insulin-resistant (P = .009), and they had higher levels of serum soluble tumor necrosis alpha receptor II (TNFalpha RII) (P = .002), triglycerides (P = .006), uric acid (P = .001), and leptin (P = .002) than did the controls. However, the study groups did not differ in serum TNFalpha, C-reactive protein (CRP), interleukin-6 (IL-6), sex hormone-binding globulin (SHBG), and high density lipoprotein-2 (HDL(2))-cholesterol. In multiple regression analysis only SHBG (P = .01) and triglycerides (P = .0036) were associated with insulin sensitivity independently of body mass index (BMI), weight gain, HDL(2) cholesterol, CRP, TNFalpha, and TNFalpha RII, IL-6, and leptin. We conclude that insulin resistance and the inflammatory markers studied were not associated in established preeclampsia. PMID- 15536599 TI - Stimulatory short-term effects of free fatty acids on glucagon secretion at low to normal glucose concentrations. AB - While free fatty acids (FFA) are well known as insulin secretagogues, their effects on pancreatic alpha cells have been mostly neglected. In the present study we therefore systematically analyzed the glucagon metabolism of rat pancreatic islets under the influence of FFA. Primary islets were incubated in the presence or absence of 200 micromol/L albumin-complexed palmitate or oleate at 2.8 mmol/L versus 16.7 mmol/L glucose and glucagon secretion was monitored over 8 hours. In addition to these time-course experiments, dose dependency of palmitate-induced effects was tested by a 2-hour incubation with 50 to 300 micromol/L albumin-complexed palmitate at 2.8 mmol/L and 5.6 mmol/L glucose. Apart from glucagon secretion, intracellular immunoreactive glucagon and cellular preproglucagon-mRNA (PPG-mRNA) content were determined from the remaining cell lysates. FFA, especially palmitate, induced a significant and dose-dependent increase of glucagon secretion (in average 2-fold above control) during the first 120 minutes of incubation at low to normal glucose (2.8 and 5.6 mmol/L). There was no significant glucagonotropic effect of FFA at concomitant 16.7 mmol/L glucose. Intracellular glucagon as well as cellular PPG-mRNA content were found to be dose-dependently diminished by palmitate when compared with untreated controls at 5.6 mmol/L glucose. The present analysis therefore points to a new role for FFA as a nutritient secretagogue and a modulator of alpha-cellular glucagon metabolism. PMID- 15536600 TI - Preserved circadian rhythm of serum insulin concentration at low plasma glucose during fasting in lean and overweight humans. AB - Circadian rhythms in glucose metabolism are well documented. Most studies, however, evaluated such variations under conditions of continuous glucose supply, either via food intake or glucose infusion. Here we assessed in 30 subjects circadian variations in concentrations of plasma glucose, serum insulin, and C peptide during a 72-hour fasting period to evaluate rhythms independent from glucose supply. Furthermore we assessed differences in these parameters between normal-weight (n = 20) and overweight (n = 10) subjects. Blood was sampled every 4 hours. During fasting, plasma glucose, serum insulin, and C-peptide levels gradually decreased (all P < .001). While there was no circadian variation in plasma glucose levels after the first day of fasting, serum levels of insulin were constantly higher in the morning (8.00 h) than at night (0.00 h) (P < .001), although the extent of this morning-associated rise in insulin levels decreased with the time spent fasting (P = .001). Also, morning C-peptide concentrations were higher compared to the preceding night (P < .001). The C-peptide/insulin ratio (CIR) decreased during prolonged fasting (P = .030), suggesting a decrease in hepatic insulin clearance. Moreover, CIR was significantly lower in the morning than at the night of day 1 and day 2 of fasting (P = .010 and P = .004, respectively). Compared to normal-weight subjects, overweight subjects had higher plasma glucose, as well as serum insulin and C-peptide levels (all P < .03). Data indicate preserved circadian rhythms in insulin concentrations in the presence of substantially decreased glucose levels in normal-weight and overweight subjects. This finding suggests a central nervous system contribution to the regulation of insulin secretion independent of plasma glucose levels. PMID- 15536601 TI - Adiponectin and C-reactive protein in obesity, type 2 diabetes, and monodrug therapy. AB - To learn more about the factors that regulate adipokines in diabetes, we examined fasting plasma concentrations of adiponectin and C-reactive protein (CRP) in well characterized groups of age-matched individuals classified as: (1) type 2 diabetes; (2) impaired fasting glucose or mild diabetes (IFG/mild DM); (3) obese, matched for body mass index (BMI); and (4) non-obese. Diabetic subjects were also studied on no phamacologic treatment, after 3 months randomization to metformin or glyburide, and after 3 months crossover to the opposite drug. CRP decreased and adiponectin increased progressively between subjects in groups 1 through 4. CRP was significantly associated with percent (r = 0.45) and total (r = 0.50) fat, insulin sensitivity as S(I) (r = -0.39) or homeostasis model assessment of insulin resistance [HOMA (IR)] (r = -0.36), and hemoglobin A(1c) (HbA(1c)) (r = 0.41). The relationship of CRP to percent fat appeared to be logarithmic and log CRP varied with percent fat independent of gender. Adiponectin concentration was significantly associated with insulin sensitivity as S(I) (r = 0.55) or HOMA (IR) (r = -0.46). Adiponectin concentrations were higher among women overall (all groups included) but not in women classified as type 2 diabetes. Although mean adiponectin was higher in subjects classified as non-obese compared to obese, adiponectin, in sharp contrast to leptin (previously reported data) and to CRP, varied markedly when expressed as a function of adiposity. Multiple regression models confirmed the strong relationship of adiponectin to insulin sensitivity, as well as the relationships of CRP to adiposity and insulin sensitivity. Glyburide treatment of diabetes decreased CRP and did so even though body weight increased. We conclude that both CRP and adiponectin correlate strongly to S(I). CRP, in contrast to adiponectin, is far more dependent on adiposity. The relationship between CRP (like leptin) and gender depends on how CRP is expressed relative to adiposity. Our data raise the possibility that gender differences in adiponectin may be lost in diabetes. Finally, pharmacologic treatment of diabetes may modulate CRP independent of adiposity. PMID- 15536602 TI - Analysis of the relationship between body mass index, insulin resistance, and beta-cell function: a cross-sectional study using the minimal model. AB - The objective of our research was to identify the mathematical model that would best define the relationship between obesity, insulin resistance (IR), and beta cell function. Eighty-seven healthy subjects with a wide range of body mass index (BMI) were studied. Insulin sensitivity (IS) was calculated using Bergman's minimal model. Acute insulin response (AIRg) was calculated as the secretion of insulin during the first 10 minutes following a glucose bolus. IS x AIRg was used as an index of insulin-mediated glucose uptake (IMGU). The relationships among BMI, IS, fasting plasma insulin (FPI), and AIRg were studied in linear relationship terms and in terms of the hyperbolic function. Where the best fit was linear, the Jones and Molitoris method was used to investigate whether the 2 line fit was significantly better. The division of the population into BMI quartiles shows that from the third quartile, IS (12.4 +/- 6.0 v 11.0 +/- 6.4 v 4.8 +/- 1.8 v 3.2 +/- 2.0 E-5 min(-1)[pmol/L](-1), P < .01) diminishes. Nevertheless, a plateau was established between the last 3 quartiles for IS x AIRg. AIRg related to BMI via a breakpoint of 29.3 kg . m(-2). The best fits for both the BMI/IS and BMI/FPI relationships were hyperbolic. Our data indicate that obesity represents a continuum of IR, with severity increasing as BMI increases. Nevertheless, above a value of 29 kg . m(-2) and despite great increases in adiposity, IS tends to descend slowly. Moreover, there seems to be an IMGU threshold at a BMI value of approximately 27 kg . m(-2), above which an increase in adiposity leads to a greater fall in IS x AIRg. Furthermore, this threshold also appears to affect pancreatic response to a glucose stimulus. PMID- 15536603 TI - Insulin regulation of free fatty acid kinetics in adult cystic fibrosis patients with impaired glucose tolerance. AB - Cystic fibrosis (CF) patients are insulin-resistant with regards to suppression of hepatic glucose production and proteolysis, but the effect of insulin on adipose free fatty acid (FFA) release has not been studied. [9,10-(3)H]palmitate kinetics were measured in 11 stable adult CF patients with impaired glucose tolerance (IGT) and 9 normal control subjects. Baseline plasma palmitate concentrations [CF = 99 +/- 13 (median 74, range 65 to 187); control = 88 +/- 9 (88, 46 to 138) micromol/L, P = .9] and palmitate flux [CF = 114 +/- 11 (100, 72 to 171); control = 105 +/- 12 (106, 54 to 182) micromol/min, P = 0.9] were not different between CF patients and controls. During a euglycemic clamp with infusion of insulin to physiologic postprandial levels, however, palmitate concentrations tended to be higher in CF patients: CF = 18 +/- 3 (13, 10 to 47), control = 12 +/- 1 (11, 8 to 18) micromol/L, P = 0.08. The higher palmitate concentrations during hyperinsulinemia appeared to be due to reduced suppression of adipose tissue palmitate release, because mean palmitate flux was 33% greater in CF subjects [32 +/- 5 (26, 17 to 66) micromol/min] than controls: [24 +/- 2 (23, 17 to 34) micromol/min], P = .20. There was considerably greater heterogeneity in insulin-induced suppression of plasma palmitate concentration and flux in CF patients compared to normal control subjects. In summary, a defect in insulin suppression of lipolysis was seen in clinically stable CF patients with IGT, similar to what has been described in CF for amino acid and glucose metabolism. This quantitative difference in lipolysis may account for inadequate insulin-induced suppression of hepatic glucose production in CF, and may be a metabolic adaptation to increased energy needs. PMID- 15536604 TI - Effect of pioglitazone on the early stage of type 2 diabetic nephropathy in KK/Ta mice. AB - Pioglitazone (PIO) has preventive effects on impaired glucose tolerance (IGT) and urinary albumin excretion in diabetes. These effects in the early stage of diabetic nephropathy have not been fully described. Endothelial constitutive nitric oxide synthase (ecNOS) might be one of the mechanisms of glomerular hyperfiltration. The objective of the present study was to evaluate the effect of PIO, including the role of ecNOS on the early stage of diabetic nephropathy in KK/Ta mice. KK/Ta mice were given PIO (10 mg/kg/d) started at 12 or 16 weeks of age for 8 or 4 weeks, respectively. They were divided into 3 groups as follows: early treatment (n = 8), late treatment (n = 8), and control group (n = 12). The urinary albumin/creatinine ratio (ACR), fasting and casual blood glucose levels, ratio of glomerular and Bowman's capsule volume (GB ratio), and systemic blood pressure were measured as phenotypic characterizations. The ecNOS and iNOS protein expression in glomeruli were evaluated by immunofluorescence. PIO, especially early treatment, improved the ACR and the GB ratio, and ecNOS protein expression was decreased in the endothelium of glomerular vessels. The iNOS protein was not detectable. There were no significant changes in the levels of fasting and casual blood glucose and systemic blood pressure among all groups. We conclude that the effect of PIO on microalbuminuria might not be due to changing systemic blood pressure and blood glucose levels. It appears that the decrease of urinary albumin excretion might be related to improvement of glomerular enlargement, including hyperfiltration, since the levels of ecNOS protein were reduced by PIO in the glomerular vessels. PMID- 15536605 TI - Homocysteine remethylation and trans-sulfuration. AB - The plasma homocysteine (Hcy) concentration represents the balance between its entry and removal from the circulation. This understanding has stimulated efforts to elucidate the causes of hyperhomocysteinemia by measuring plasma Hcy turnover. However, these studies have been performed under steady-state conditions, which do not allow for conclusions about the type and severity of the metabolic blocks that cause metabolites to accumulate. Failure to appreciate this has led to some confusion in the literature dealing with whole body Hcy metabolism. PMID- 15536606 TI - Interstitial glucose kinetics in subjects with type 1 diabetes under physiologic conditions. AB - We investigated the dynamic relationship between interstitial glucose (IG) in the subcutaneous adipose tissue and plasma glucose (PG) during physiologic conditions in type 1 diabetes mellitus (T1DM). Nine subjects with T1DM (5/4 M/F; age, 33 +/- 13 years; body mass index, 26.6 +/- 4.3 kg/m(2); glycosylated hemoglobin [HbA(1c)], 8.6% +/- 0.9%; mean +/- SD) treated by continuous subcutaneous insulin infusion (CSII) with insulin lispro were studied over 12 hours after a standard meal (40 g carbohydrate [CHO]) and prandial insulin. IG was measured by open flow microperfusion. Nine compartment models were postulated to account for temporal variations in the IG/PG ratio. The models differed in the inclusion of physiologically motivated alterations of pathways entering/leaving the IG compartment in the adipose tissue. The best model included zero order (constant) glucose disposal from the interstitial fluid (ISF) and insulin-stimulated glucose transfer from plasma to the ISF. The former effect is expressed by a positive association between the IG/PG ratio and PG, eg, a decrease in PG from 9 to 3.3 mmol/L lowers the IG/PG ratio by 0.1. The latter effect results in the IG/PG ratio to be increased by 0.03 per 10 mU/L of plasma insulin. We were not able to detect the stimulatory effect of insulin on glucose disappearance from the ISF. In conclusion, we developed and quantified a model of IG kinetics in the adipose tissue applicable to physiologic conditions in subjects with T1DM. PMID- 15536607 TI - Ionomycin, but not physiologic doses of epinephrine, stimulates skeletal muscle interleukin-6 mRNA expression and protein release. AB - It has been hypothesized that epinephrine may stimulate interleukin (IL)-6 gene expression in skeletal muscle. The aim of the present study was to examine the effect of epinephrine on IL-6 gene expression within, and protein release from, skeletal muscle. We hypothesized that physiologic epinephrine would neither result in an increase in IL-6 mRNA nor protein release from skeletal muscle. Soleus muscle was excised from 4-week-old anesthetized Sprague Dawley rats and incubated in a Krebs buffer with the addition of either saline (CON), epinephrine, at concentrations of 1,000 nmol/L (EPI 1,000), 100 nmol/L (EPI 100), or 10 nmol/L (EPI 10), or the calcium ionophore, ionomycin (IONO), a positive control. After a 1-hour incubation, muscle was collected and extracted for RNA, reverse transcribed, and IL-6 gene expression was determined by real-time polymerase chain reaction (PCR). An aliquot of incubation medium was also collected and analyzed for IL-6 protein by enzyme-linked immunosorbent (ELISA). EPI 1,000 and IONO increased (P < .05) IL-6 mRNA, whereas EPI 100 and EPI 10 were without effect. IL-6 protein release from skeletal muscle was increased in IONO (P < .05), but not in CON or EPI at any concentration. These data demonstrate that while pharmacologic concentrations of epinephrine activate IL-6 mRNA, supraphysiologic and high-physiologic doses appear to have little, if any, effect on IL-6 gene transcription in skeletal muscle. In addition, ionomycin can stimulate IL-6 gene expression and protein release after only 1 hour of exposure. PMID- 15536608 TI - Hemoglobin A1c can be analyzed in blood kept frozen at -80 degrees C and is not commonly affected by hemolysis in the general population. AB - The ability of glycated hemoglobin A(1c) (HbA(1c)) to predict diabetes is unknown, but could be evaluated by analyses on samples stored in biobanks. The stability of HbA(1c) in long-term stored samples is, however, unknown. Moreover, the effect of hemolysis on HbA(1c) in the general population is not assessed. To explore these questions HbA(1c) was determined in 3 groups (n = 717) of samples with storage times at -80 degrees C differing between 10 years and 2 months. The results were compared with HbA(1c) analyzed in fresh blood samples (n = 174). The subjects were free from diabetes and aged 40 to 60 years. HbA(1c) was analyzed by cation exchange high-performance liquid chromatography (HPLC). The mean HbA(1c) results for the fresh and long-term stored samples were 4.25% +/- 0.39 and 4.19% +/- 0.43, respectively (P = not significant [NS]). The HbA(1c) levels in fresh and 2-month stored samples were essentially equal. There was no correlation between HbA(1c) in the fresh samples and the hemolysis parameters reticulocytes, haptoglobin, or bilirubin. HbA(1c) is apparently stable in samples long-term stored at -80 degrees C and is not commonly affected by hemolysis in the general population. HbA(1c) analyzed on biobank samples could be used to assess the predictive value for future diabetes and relationship to other morbidity and mortality. PMID- 15536609 TI - Identification of a new point mutation in hypoxanthine phosphoribosyl transferase responsible for hyperuricemia in a female patient. AB - A 29-year-old woman was referred to our department because of gout. Routine laboratory data showed hyperuricemia, a high level of plasma oxypurines, increased urinary uric acid excretion, and increased urinary oxypurine excretion, with decreased hypoxanthine phosphoribosyl transferase (HPRT) activity in the erythrocytes. From these findings, the patient was diagnosed with a partial deficiency of HPRT. To determine its properties, a cDNA sequence encoding HPRT and the androgen receptor AR XIST minimal promoter gene, as well as methylation of the AR gene were investigated. The HPRT cDNA sequence revealed a point mutation of G to A in nucleotide 40, which changed codon 14 from GAA (Glu) to AAA (Lys) in the mutant gene. In addition, the HPRT genomic DNA sequence, including the mutation site, revealed the same point mutation, indicating that the patient was heterozygote. Further analysis of the AR gene on the X chromosome suggested nonrandom X-chromosome inactivation, whereas the AR XIST minimal promoter gene was normal. Such results have not been previously reported in a female with partial HPRT deficiency. PMID- 15536610 TI - Lifestyle behaviors associated with lower risk of having the metabolic syndrome. AB - The metabolic syndrome is a cluster of risk factors that predisposes individuals to cardiovascular disease (CVD) and diabetes and is present in almost one fourth of adult Americans. Risk factors involved with the metabolic syndrome can be altered via modifiable lifestyle factors, such as diet, physical activity, and smoking and drinking habits. The objective of this study was to examine the extent to which these modifiable lifestyle behaviors are associated with the risk of having the metabolic syndrome. Data from the Third National Health and Nutrition Examination Survey (NHANES III), conducted between 1988 and 1994, were used to measure the risk of having the metabolic syndrome in healthy adult Americans who follow certain lifestyle behaviors, such as dietary practices, levels of physical activity, smoking and drinking habits. Low physical activity level, high carbohydrate (CHO) intake, and current smoking habits were all significantly associated with an increased risk of having the metabolic syndrome, even after adjusting for other related covariates. Relative to physically inactive subjects, being physically active was associated with lower odds ratio (OR) (0.36, confidence interval [CI] 0.21 to 0.68, P < .01) in overweight men and in normal weight (0.36, CI 0.18 to 0.70, P < .01) and overweight (0.61, CI 0.38 to 0.97, P < .05) women. Although the type of CHO could not be distinguished, relative to a high CHO diet, men having a low or moderate CHO intake had a lower risk of having the metabolic syndrome with respective ORs of 0.41 (CI 0.24 to 0.67, P < .01) and 0.44 (CI 0.25 to 0.77, P < .01); no effect of dietary CHO was observed in women. Moderate alcohol consumption was not significantly related to the risk of having the metabolic syndrome in men, but was associated with a lower OR in women (0.76, CI 0.61 to 0.95, P < .05). Regression models indicate a reduced risk of having the metabolic syndrome when selected low-risk lifestyle factors are present in combination, particularly in subjects with body mass index (BMI) < 30 kg/m(2). According to our cross-sectional logistic models, the risk of having the metabolic syndrome is substantially lower in individuals who are physically active, nonsmoking, have a relatively low CHO intake and moderate alcohol consumption, and who maintain a BMI in the non-obese range. These observations have potentially important value for public health recommendations. PMID- 15536611 TI - Emerging cardiovascular risk factors in subclinical hypothyroidism: lack of change after restoration of euthyroidism. AB - Subclinical hypothyroidism (SH) is a frequent condition that may be associated with increased cardiovascular risk. There is current interest in determining the effect, if any, of substitutive therapy with l-thyroxine (L-T4) on cardiovascular risk factors in SH and, particularly, on those associated with emerging cardiovacular risk, such as apolipoprotein (apo) B, lipoprotein (Lp) (a), total homocysteine (t-Hcy), and C-reactive protein (CRP). Thus, the aim of this study was to assess the impact of euthyroidism restoration on these emerging risk factors in SH. Forty-two patients diagnosed with SH were consecutively recruited before treatment. These patients were treated with L-T4 for 3 to 6 months with the dose necessary to restore euthyroidism. Lp(a), fasting and postmethionine (n = 28) t-Hcy, and CRP did not change with substitutive therapy, regardless of the respective baseline values, and the decrease in apo B paralleled that of low density lipoprotein (LDL) cholesterol. Similarly, no treatment effect was observed on homocysteine or CRP in patients with thyrotropin-stimulating hormone (TSH) >10 mIU/L. Monitoring of emerging risk factors did not offer additional arguments for treating patients with SH and, thus, is not justified in their clinical management. PMID- 15536612 TI - Myotonia and muscle contractile properties in mice with SIX5 deficiency. AB - Myotonic dystrophy (DM1) is an autosomal-dominant multisystem disease characterized by progressive skeletal muscle weakness, myotonia, cataracts, cardiac arrhythmias, mild mental retardation, and endocrinopathies. Heterozygous loss of SIX5 in mice causes cataracts and cardiac conduction disease, and homozygous loss also leads to sterility and decreased testicular mass, reminiscent of DM1 in humans. The effect of SIX5 deficiency in muscle is unknown. In this study, we found that muscle contractile properties, electromyographic insertional activity, and muscle histology were normal in SIX5 deficient mice. The implications of these findings for the pathogenesis of DM1 are discussed. PMID- 15536613 TI - Lewis-Sumner syndrome and multifocal motor neuropathy. AB - We compared the clinical, electrophysiological, laboratory, and pathological features of 13 patients with Lewis-Sumner syndrome (LSS) with those of 20 patients with multifocal motor neuropathy (MMN). LSS and MMN patients have several common clinical features: age at onset, weakness in the distribution of individual peripheral nerves, mild wasting, cramps and fasciculations, partial areflexia, and frequent stepwise disease course. Cerebrospinal fluid protein level was normal or slightly elevated, but always less than 100 mg/dl. Conduction blocks are the electrophysiological hallmarks of these two neuropathies, and no differences in distribution and number of blocks were found. Contrary to MMN, lower-limb involvement at onset was frequent in LSS but extension to the upper limbs was a frequent later feature of the disease. Cranial nerve involvement was noted in 4 LSS patients during relapses and absent in all MMN patients. The major distinguishing features were the clinical and electrophysiological sensory involvement in LSS, and the lack of anti-GM1 antibodies in LSS, whereas IgM anti GM1 were found in 40% of MMN patients. Some LSS patients responded to steroid therapy, whereas this was ineffective in MMN. From these features, LSS can be considered an entity distinct from MMN, with its own clinical, laboratory, and electrophysiological characteristics, and as an intermediate link between chronic inflammatory demyelinating polyneuropathy and MMN. PMID- 15536614 TI - Needle EMG in certain uncommon clinical contexts. AB - It is the position of the AANEM that, based on the literature, there are no contraindications to needle electromyography (EMG) in patients with lymphedema or prosthetic joints. In patients with lymphedema, clinical judgment in each individual circumstance should be used in deciding whether the risk of complication is greater than the value of the information to be obtained from the needle electrode examination. PMID- 15536615 TI - Muscular amyloid angiopathy with amyloidgenic transthyretin Ser50Ile and Tyr114Cys. AB - Among patients with familial amyloid polyneuropathy (FAP), those with transthyretin Val30Met mainly show distally predominant weakness and atrophy, whereas some FAP patients, including those with transthyretin Ser50Ile and Tyr114Cys, show muscle weakness and atrophy that is dominant proximally, simulating myopathy. To clarify the cause of proximally dominant muscular atrophy in patients with FAP transthyretin Ser50Ile and Tyr114Cys, we investigated the distinctive features of muscle specimens of patients with FAP, 3 of who had Val30Met, 2 Ser50Ile, and 2 Tyr114Cys transthyretin. All specimens showed transthyretin amyloid around blood vessels and perimysium, and neurogenic denervation patterns. The amount of amyloid around the vessels was much greater in patients with FAP Ser50Ile and Tyr114Cys than in Val30Met patients. Muscular amyloid angiopathy may contribute to motor nerve injury that, in turn, may lead to amyotropic changes in patients with FAP Ser50Ile and Tyr114Cys. PMID- 15536616 TI - Myocytes from congenital myotonic dystrophy display abnormal Na+ channel activities. AB - Na(+) currents were measured in myocytes from a fetus with congenital myotonic dystrophy type 1 (DM1) using the patch-clamp whole-cell technique. Steady-state activation and inactivation properties of Na(+) channels were not substantially different between these cells and age-matched control cells. However, a decrease in Na(+) channel density and a faster rate of recovery from inactivation were found in myocytes from congenital DM1 suggesting that changes in functional Na(+) channels may affect cell excitability of muscle cells of patients with this disorder. PMID- 15536617 TI - Impact of functional support on health-related quality of life in patients with colorectal cancer. AB - BACKGROUND: It has been shown that social integration and the availability of social support influence quality of life. However, little is known about the relation between social support and mental and physical health in patients with colorectal cancer. In the current study, the authors examined the effects of social network size, as well as emotional and instrumental support, on health related quality of life (HRQOL) in patients with colorectal cancer. METHODS: Six hundred thirty-six veterans with colorectal cancer were asked to complete a telephone interview, which included a measure of social support (the Berkman-Syme Index) and the Medical Outcomes Study Short Form 12-Item Survey. Mean physical composite scale (PCS) and mental composite scale (MCS) scores were compared across groups. RESULTS: No difference in mean PCS or MCS scores was found between patients who had larger social networks and patients who had smaller social networks. The availability of emotional and instrumental support was associated with higher MCS scores, whereas the availability of instrumental support was associated with lower PCS scores. CONCLUSIONS: Irrespective of network size, the availability of emotional support and instrumental support had an impact on HRQOL in patients with colorectal cancer. More emphasis needs to be placed on understanding how various types of social support, individually and collectively, influence physical and mental health in patients with colorectal cancer. PMID- 15536618 TI - alpha1-acid glycoprotein fucosylation as a marker of carcinoma progression and prognosis. AB - BACKGROUND: Serum alpha1-acid glycoprotein (AGP), an acute-phase protein secreted by the liver, carries alpha(1,3)-fucosylated structures on its 5 highly branched, N-linked sugar chains. METHODS: Serum AGP levels in patients with various types of malignancies (n=214 patients) were measured using an enzyme-linked immunosorbent assay with anti-AGP antibody. To investigate glycoforms that differed in their degree of branching and extent of fucosylation, serum AGP samples were analyzed by crossed affinoimmunoelectrophoresis (CAIE) with concanavalin A, and Aleuria aurantia lectin (AAL), and anti-AGP antibody. RESULTS: A significant difference (P <0.001) in serum AGP levels was observed in preoperative patients compared with levels in the healthy control group, but the levels in individual patients did not reflect their clinical status. Conversely, it was found not only that the patterns of AGP glycoforms differed widely in the patient group compared with the healthy control group, but they also changed depending on each patient's clinical status. Furthermore, AGP glycoforms seemed to be appropriate markers of disease progression and prognosis according to follow-up studies of 45 patients during prolonged preoperative and postoperative periods. CONCLUSIONS: Patients with advanced malignancies who had AGP glycoforms that contained highly fucosylated triantennary and tetraantennary sugar chains for long periods after surgery were likely to have a poor prognosis. However, patients who had AGP glycoforms without such changes were expected to have a good prognosis. PMID- 15536619 TI - Longitudinal study of smoking patterns in relation to the development of smoking related secondary primary tumors in patients with upper aerodigestive tract malignancies. AB - BACKGROUND: The authors set out to assess the correlation between smoking-related second primary tumor (SPT) development and cigarette smoking habits after diagnosis and definitive treatment in patients with early-stage head and neck squamous cell carcinoma who were enrolled in a placebo-controlled randomized chemoprevention trial of 13-cis-retinoic acid. METHODS: Longitudinal data collected for 10 years after the index diagnosis are presented for 1190 patients. Cox proportional hazards regression models were used to examine the effects of changes in smoking behavior on smoking-related SPT development. RESULTS: One third of all patients who quit smoking within 12 months before randomization experienced recurrence, compared with 6.9% and 10.4% of all never-smokers and former smokers, respectively. Approximately 16% of all current smokers stopped smoking, and nearly 22% of current smokers developed SPTs, compared with 14.5%, 13.2%, and 8.8% of all recent smokers, former smokers, and never-smokers, respectively. The probability of developing a smoking-related SPT was highest among patients who were current smokers at randomization. These patients, regardless of whether they ceased smoking during follow-up, were nearly three times more likely than patients who had never smoked to develop a smoking-related SPT. In contrast, former smokers and recent quitters who continued to abstain from smoking during follow-up were approximately 1.5 times more likely to develop an SPT compared with patients who had never smoked. CONCLUSIONS: Patients who continue to smoke after the successful treatment of their index head and neck malignancies have a substantially higher risk of developing smoking-related SPTs. PMID- 15536620 TI - Differential diagnosis of chronic dysimmune demyelinating polyneuropathies with and without anti-MAG antibodies. AB - The distinction between chronic demyelinating polyneuropathies associated with IgM paraproteinemia and anti-myelin-associated glycoprotein (MAG) antibodies (MAG PN) and chronic inflammatory demyelinating polyneuropathies (CIDPs) relies on the anti-MAG antibodies assay. The aim of the study was to identify clinical and electrophysiological features suggesting a diagnosis of MAG-PN. Fourteen patients with MAG-PN and 35 with CIDP were included, and a discriminant analysis was performed to identify the clinical and electrophysiological features suggestive of MAG-PN. Pure sensory clinical phenotype, low median and ulnar terminal latency index, and absence of M responses in the lower limbs were significantly associated with the diagnosis of MAG-PN, and indicate a moderate to large increase in probability of this diagnosis in patients with chronic dysimmune demyelinating polyneuropathies. PMID- 15536621 TI - Rhabdomyosarcoma: value of myogenin expression analysis and molecular testing in diagnosing the alveolar subtype: an analysis of 109 paraffin-embedded specimens. AB - BACKGROUND: Identification of the alveolar subtype of rhabdomyosarcoma (ARMS) is important, because the poor prognosis associated with this subtype necessitates a modified therapeutic regimen. At present, ARMS diagnoses are made on the basis of histologic findings and the extent of myogenin immunopositivity. Nonetheless, the absence of an alveolar pattern in the solid variant, the low degree of differentiation in certain embryonal rhabdomyosarcomas (ERMS), and the increasing use of microbiopsy samples make the diagnosis of ARMS somewhat difficult. Two specific translocations have been found in ARMS, and fusion transcripts can be detected by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of paraffin-embedded tissue (PET). METHODS: To assess the value of myogenin staining and molecular testing in the diagnosis of rhabdomyosarcoma, the authors examined 109 rhabdomyosarcoma samples (45 ARMS samples and 64 ERMS samples). Real-time RT PCR analysis of PET was performed in all 109 rhabdomyosarcomas, and RT-PCR analysis of frozen material was performed in 24 cases. RESULTS: PAX fusion transcripts were present in 44 cases (39 ARMS and 5 ERMS) and absent in 52 cases (2 ARMS and 50 ERMS). In 13 cases (4 ARMS and 9 ERMS), the results were not interpretable. Results were concordant between paired frozen and fixed tumor samples. All 35 interpretable ERMS samples that contained < 50% myogenin-positive cells failed to yield detectable PAX fusion transcripts. Of the 61 interpretable tumor samples (41 ARMS and 20 ERMS) that contained > 50% myogenin-positive cells, 44 (39 ARMS and 5 ERMS) yielded detectable PAX fusion transcripts. CONCLUSIONS: The current study demonstrates that molecular detection of PAX fusion transcripts via real-time RT-PCR analysis of PET is a sensitive and specific method for the diagnosis of ARMS and that immunohistochemical analysis of myogenin expression can be used to select cases for such molecular testing. Although RT-PCR analysis appears not to possess diagnostic value in tumors with < 50% tumor cell immunopositivity, it is strongly recommended for the diagnosis of tumors containing > 50% myogenin-positive cells. PMID- 15536622 TI - Effects of evoked pain on the electromyogram and compound muscle action potential of the brachial biceps muscle. AB - Muscle pain is often accompanied by a feeling of muscle fatigue and weakness. We examined the effect of experimental muscle pain on the electromyogram (EMG) during maximal voluntary contraction (MVC), and on the compound muscle action potential (CMAP) of the brachial biceps muscle. Twenty-one healthy subjects were injected intramuscularly with 0.1 ml/30 microg of the vanilloid receptor agonist (capsaicin) or 0.2 ml of 5% hypertonic saline. A Teflon-coated cannulated EMG needle was used to record the EMG interference pattern (IP) at MVC. The CMAP of the brachial biceps muscle was obtained by stimulation of the musculocutaneous nerve at the axilla using surface electrodes. Amplitude, mean frequency of the power spectrum, and turns/s of the interference pattern were reduced after pain induced by capsaicin or hypertonic saline. Latency, amplitude, and area-under curve of the CMAP did not change after injection of either substance. Acute stimulation of muscle nociceptors thus produced a fatiguelike change in the interference pattern during MVC, possibly due to a decrease in motoneuron firing rate and increased muscle fatigability. PMID- 15536623 TI - Human ovarian carcinoma cells: histone deacetylase inhibitors exhibit antiproliferative activity and potently induce apoptosis. AB - BACKGROUND: Histone deacetylase inhibitors (HDACIs) can inhibit proliferation, stimulate apoptosis, and induce cell cycle arrest in malignant cells. METHODS: The authors investigated the effects of four HDACIs on nine ovarian carcinoma cell lines in vitro and in vivo. Ovarian carcinoma cells were treated with a variety of HDACIs, and their effects on cell growth, the cell cycle, apoptosis, and related events were investigated. The ability of valproic acid (VPA) to inhibit the growth of ovarian tumors in immunodeficient mice was also assessed. RESULTS: Clonogenic assays showed that all ovarian carcinoma cell lines were sensitive to the growth-inhibitory effects of the HDACIs. Cell cycle analysis indicated that their exposure to HDACIs decreased the proportion of cells in S phase and increased the proportion of cells in the G0/G1 and/or G2/M phases of the cell cycle. Terminal deoxynucleotidyltransferase-mediated uridine triphosphate end-labeling assays demonstrated that HDACIs induced apoptosis, which occurred in concert with alterations in the expression of genes related to apoptosis, cell growth, and malignant phenotype, including the activation of caspase-9 and caspase-3. Chromatin immunoprecipitation analysis revealed a notable increase in levels of acetylated histones associated with the p21 promoter after treatment with suberoylanilide bishydroxamine. In addition, in experiments involving nude mice, VPA significantly inhibited human ovarian tumor growth without toxic side effects. CONCLUSIONS: The results of the current study suggest that HDACIs may be particularly effective in the treatment of ovarian tumors. PMID- 15536624 TI - Distal symmetrical polyneuropathy: definition for clinical research. AB - The objective of this report was to develop a case definition of "distal symmetrical polyneuropathy" to standardize and facilitate clinical research and epidemiological studies. A formalized consensus process was employed to reach agreement after a systematic review and classification of evidence from the literature. The literature indicates that symptoms alone have relatively poor diagnostic accuracy in predicting the presence of polyneuropathy; signs are better predictors of polyneuropathy than symptoms; and single abnormalities on examination are less sensitive than multiple abnormalities in predicting the presence of polyneuropathy. The combination of neuropathic symptoms, signs, and electrodiagnostic findings provides the most accurate diagnosis of distal symmetrical polyneuropathy. A set of case definitions was rank ordered by likelihood of disease. The highest likelihood of polyneuropathy (useful for clinical trials) occurs with a combination of multiple symptoms, multiple signs, and abnormal electrodiagnostic studies. A modest likelihood of polyneuropathy (useful for field or epidemiological studies) occurs with a combination of multiple symptoms and multiple signs when the results of electrodiagnostic studies are not available. A lower likelihood of polyneuropathy occurs when electrodiagnostic studies and signs are discordant. For research purposes, the best approach for defining distal symmetrical polyneuropathy is a set of case definitions rank ordered by estimated likelihood of disease. The inclusion of this formalized case definition in clinical and epidemiological research studies will ensure greater consistency of case selection. PMID- 15536625 TI - Thromboembolic events with estramustine phosphate-based chemotherapy in patients with hormone-refractory prostate carcinoma: results of a meta-analysis. AB - BACKGROUND: Combinations of cytotoxic chemotherapy with estramustine phosphate (EMP), a nornitrogen mustard-estrogen conjugate, are used to treat patients with hormone-refractory prostate carcinoma (HRPC). However, thromboembolic events (TE), including deep venous thrombosis (DVT), pulmonary embolism, stroke, myocardial infarction, and arterial thrombosis, are significant toxicities of these regimens. The current study sought to establish the rate of TE and to determine risk factors for TE. METHODS: A MEDLINE-based search identified EMP based clinical trials published in the English-language peer-reviewed literature after 1990 in which > or = 20 patients with HRPC were enrolled and TE were clearly documented. Patient characteristics and the dose of EMP given were analyzed to determine their association with the rate of TE. RESULTS: Twenty three studies, enrolling a total of 896 patients, were included in the analysis. The overall risk of TE was 0.07 (95% confidence interval [95% CI], 0.05-0.11). The risk of DVT was 0.06 (95% CI, 0.04-0.09). The risks of all other types of TE were <0.01. Using univariate logistic regression analysis, the dose of EMP administered, baseline patient age, and baseline prostate-specific antigen level were not found to be associated with the total risk of TE. The rates of total TE and DVT may be inflated because one of the analyzed studies initially had a very high rate of DVT (25%) when compared with the others. CONCLUSIONS: The rate of TE in men with HRPC who are treated with EMP-based regimens is significant, but it does not appear to be related to the dose of EMP. Whether TE can be prevented with anticoagulant prophylaxis remains to be determined. PMID- 15536626 TI - Fragmentation of N-linked glycans with a matrix-assisted laser desorption/ionization ion trap time-of-flight mass spectrometer. AB - N-Linked glycans were ionized from several matrices with a Shimadzu-Biotech AXIMA QIT matrix-assisted laser desorption/ionization quadrupole ion trap time-of flight mass spectrometer. [M+Na]+ ions were produced from all matrices and were accompanied by varying amounts of in-source fragmentation products. The least fragmentation was produced by 2,5-dihydroxybenzoic acid and the most by alpha cyano-4-hydroxycinnamic acid and 6-aza-2-thiothymine. Sialic acid loss was extensive but could be prevented by formation of methyl esters. Fragmentation produced typical low-energy-type spectra dominated by ions formed by glycosidic cleavages. MS(n) spectra (n = 3 and 4) were used to probe the pathways leading to the major diagnostic ions. Thus, for example, an ion that was formed by loss of the core GlcNAc residues and the 3-antenna was confirmed as being formed by a B/Y rather than a C/Z mechanism. The proposed structures of several cross-ring cleavage ions were confirmed and it was shown that MS3 spectra could be obtained from as little as 10 fmol of glycan. PMID- 15536627 TI - Site-specific carbohydrate profiling of human transferrin by nano-flow liquid chromatography/electrospray ionization mass spectrometry. AB - Glycopeptides derived from a lysylendopeptidase digest of commercially available human transferrin were analyzed by nano-flow liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS), which permitted the carbohydrate profiles at Asn432 and Asn630 to be determined. Both are located in a well-known motif for N-glycosylation, Asn-Xaa-Ser/Thr. The contents of the carbohydrates at each site were significantly different from each other, and consisted of a variety of minor types of oligosaccharides in addition to the major one, a biantennary complex-type oligosaccharide. Nano-flow ESI tandem mass spectrometry (MS/MS) of the glycopeptides (Cys421-Lys433 and Ile619-Lys646) containing these two sites yielded predominantly ions originating from the non-reducing termini (oxonium ions) and reducing terminus, resulting from cleavage of the glycosidic bonds of the carbohydrate moieties; this permitted the structural read-out of a small minority of the carbohydrate moieties. In particular, the observation of oxonium ions at m/z 512.2 and 803.2 is useful for probing outer non-reducing terminal fucosylation, which represented carbohydrate structures consisting of Hex, dHex, and HexNAc, and NeuNAc, Hex, dHex, and HexNAc, respectively, from which the Lewis X structure (Galbeta1-4(Fucalpha1-3)GlcNAc) was readily deduced. Moreover, fucosylation at the reducing-terminal GlcNAc (Fucalpha1-6GlcNAc) specifically occurred at Asn630, as demonstrated by treatment of the glycopeptides with alpha1-3/4-L-fucosidase. PMID- 15536628 TI - Production and collision-induced dissociation of gas-phase, water- and alcohol coordinated uranyl complexes containing halide or perchlorate anions. AB - Electrospray ionization was used to generate mono-positive gas-phase complexes of the general formula [UO2A(S)n]+ where A = OH, Cl, Br, I or ClO4, S = H2O, CH3OH or CH3CH2OH, and n = 1-3. The multiple-stage dissociation pathways of the complexes were then studied using ion-trap mass spectrometry. For H2O-coordinated cations, the dissociation reactions observed included the elimination of H2O ligands and the loss of HA (where A = Cl, Br or I). Only for the Br and ClO4 versions did collision-induced dissociation (CID) of the hydrated species generate the bare, uranyl-anion complexes. CID of the chloride and iodide versions led instead to the production of uranyl hydroxide and hydrated UO2+. Replacement of H2O ligands by alcohol increased the tendency to eliminate HA, consistent with the higher intrinsic acidity of the alcohols compared to water and potentially stronger UO2-O interactions within the alkoxide complexes compared to the hydroxide version. PMID- 15536629 TI - Removal of sodium and potassium adducts using a matrix additive during matrix associated laser desorption/ionization time-of-flight mass spectrometric analysis of peptides. AB - Monovalent cations often associate with peptides and proteins under mass spectrometry (MS) conditions, resulting in a discernable, but often misleading, adduct cluster pattern. These adduct cluster peaks reduce the signal intensity of specific peptide species by splitting the ion population into multiple mass peaks, suppressing the ionization of neighboring low-abundance peaks, and interfering with identification of post-translational modifications. Further, monovalent contaminants tend to form a distribution of matrix cluster peaks in matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) spectra causing interference and suppression in the mass range below 1400 Da. The most common method for reduction or elimination of adduct clusters is solid-phase extraction via a pipette tip or spin column, which often leads to loss of low abundance peptide components. In this study we describe the use of a commercially available surfactant blend that markedly reduces the adduction of monovalent cations during peptide analysis by MALDI-TOFMS. PMID- 15536630 TI - Isotope-coded N-terminal sulfonation of peptides allows quantitative proteomic analysis with increased de novo peptide sequencing capability. AB - Recently various methods for the N-terminal sulfonation of peptides have been developed for the mass spectrometric analyses of proteomic samples to facilitate de novo sequencing of the peptides produced. This paper describes the isotope coded N-terminal sulfonation (ICenS) of peptides; this procedure allows both de novo peptide sequencing and quantitative proteomics to be studied simultaneously. As N-terminal sulfonation reagents, 13C-labeled 4-sulfophenyl[13C6]isothiocyanate (13C-SPITC) and unlabeled 4-sulfophenyl isothiocyanate (12C-SPITC) were synthesized. The experimental and reference peptide mixtures were derivatized independently using 13C-SPITC and 12C-SPITC and then combined to generate an isotopically labeled peptide mixture in which each isotopic pair differs in mass by 6 Da. Capillary reverse-phase liquid chromatography/tandem mass spectrometry experiments on the resulting peptide mixtures revealed several immediate advantages of ICenS in addition to the de novo sequencing capability of N terminal sulfonation, namely, differentiation between N-terminal sulfonated peptides and unmodified peptides in mass spectra, differentiation between N- and C-terminal fragments in tandem mass spectra of multiply protonated peptides by comparing fragmentations of the isotopic pairs, and relative peptide quantification between proteome samples. We demonstrate that the combination of N terminal sulfonation and isotope coding in the mass spectrometric analysis of proteomic samples is a viable method that overcomes many problems associated with current N-terminal sulfonation methods. PMID- 15536631 TI - Coupling of nanoflow liquid chromatography to matrix-assisted laser desorption/ionization mass spectrometry: real-time liquid chromatography run mapping on a MALDI plate. AB - The major obstacle in the use of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) instruments in the analysis of complex proteome samples is the lack of a direct coupling of a highly resolving separation technique with the mass spectrometer itself. To overcome this drawback, a spotting device for capillary and nanoflow liquid chromatography (LC) with a special liquid deposition principle for lowest volumes was developed. The instrument is able to perform MALDI spotting in real time in order to deposit the LC run on the MALDI plate, and therefore couples the high resolution power of nano-RP-HPLC separation directly with MALDI-MS. This work describes the development and optimization of a method for spotting with online matrix addition, and illustrates its use in the analysis of a complex proteome sample. PMID- 15536632 TI - A liquid-phase microextraction method, combining a dual gauge microsyringe with a hollow fiber membrane, for the determination of organochlorine pesticides in aqueous solution by gas chromatography/ion trap mass spectrometry. AB - A liquid-phase microextraction (LPME) method has been demonstrated for the extraction and determination of organochlorine pesticides (OCPs) in aqueous solution. The method combines a dual gauge microsyringe with a hollow fiber membrane (LPME/DGM-HF) followed by detection by gas chromatography/ion trap mass spectrometry (GC/ITMS). The advantages include speed, low solvent and sample consumption, simplicity and ease of use. The extraction time, solvent selection, salt concentration and sample stirring rate have been investigated in order to optimize extraction efficiency. The viability is evaluated by measuring the linearity and detection limit of the five OCPs in aqueous solution. Detection linearity for the OCPs has been achieved over a range of concentrations between 1 and 500 microg/L (r2 > 0.930), with a detection limit of 0.1 microg/L for each OCP. PMID- 15536633 TI - Electrospray tandem mass spectrometry of 2H-chromenes. AB - Several 2H-chromenes derived from carbazoles were analyzed by electrospray tandem mass spectrometry. The 2H-chromenes constitute an important class of compounds that exhibit photochromic activity. The fragmentation pathways of the protonated molecular species [M+H]+ were studied, and main fragmentation pathways of these compounds were identified. Fragmentation pathways of [M+D]+ ions were also studied in order to obtain information about the location of the ionizing proton or deuteron. It was found that the proton is not preferentially located on the nitrogen atom. The charge is preferentially located as a tertiary carbocation, resulting from the uptake of the proton (or deuteron) by the zwitterionic open structure of the chromenes. The major fragmentation occurred by cleavage of the gamma-bond relative to the carbocation center, leading to a fragment at m/z 191 (C5H11+ or C14H9N+), which are the most abundant fragment ions for almost all compounds. The presence of substituents in the chromene ring does not change this behavior. Other observed common fragmentation pathways included loss of CH3* (15 Da), loss of CO (28 Da), combined loss of CO and CH3 (43 Da), and loss of the phenyl ring via combined loss of C6H4 and CH3* (-91 Da) and combined loss of C6H6 and CO (-106 Da). PMID- 15536634 TI - Heterogeneous cell response to topotecan in a CFSE-based proliferation test. AB - BACKGROUND: Carboxyfluorescein diacetate succinimidyl ester (CFSE) is currently used to investigate migration and proliferation of hemopoietic cells. In principle, CFSE is retained by the cells and is shared by the daughter cells at each division, resulting in multimodal flow cytometric CFSE histograms, with each cell generation clustering around half the fluorescence intensity of the previous one. However, intercell variability of CFSE loading results in overlapping peaks, thereby limiting its use with cancer cell lines. METHODS: We used IGROV1 ovarian cancer cells loaded with CFSE at the time of seeding; 24 h later cells were treated with an anticancer drug (topotecan). Potential pitfalls of the analysis were examined, and a procedure of evaluation of CFSE efflux was applied to fix the peak positions with good approximation in advance. Histograms were fitted by a series of gaussians, with each representing cells in a given generation. RESULTS: Effects of topotecan on IGROV1 cells were analyzed in terms of the time course of the percentage of cells that remained undivided or entered the second, third, and subsequent division cycles. A simple algorithm, which combined flow cytometric data with the absolute cell number independently measured by Coulter counter, provided an estimate of the 96-h outcome of the starting cell population by quantifying cells that remained undivided, those able to divide at least once, or those that had died. CONCLUSIONS: We assessed experimental and data analytic procedures for a CFSE-based measurement of antiproliferative activity of drugs in cancer cell lines. A quantitative level was achievable but required a strict procedure for control of the experimental data, which was not straightforward. PMID- 15536635 TI - Partial characterization of glycosphingolipids of Agelas sponges in their peracetylated form by liquid secondary ionization mass spectrometry and high performance liquid chromatography combined with direct electrospray ionization mass spectrometric detection. AB - Electrospray ionization (ESI) and liquid secondary ionization (LSI) mass spectrometry were applied for characterization of glycosphingolipids (GSLs) isolated in their peracetylated form from four Agelas marine sponge species. Since peracetylated glycosphingolipids are not soluble in solvents traditionally used for ESI, lithium chloride was added to the samples in order to obtain lithium cationized molecules. Although the preferred fragmentation seems to be the sequential loss of acetic acid molecules, it was found that tandem mass spectra obtained from peracetylated diglycosyl ceramides might provide direct information about the structure of the long-chain base (formation of W''/Z0 fragments). The utility of ESI and LSI in the analysis of these compounds has also been compared. It was found that the tandem mass spectra obtained by LSI MS/MS experiments could provide information about the chain-length (carbon atom number) variations within a certain ceramide mass. Thus, from one of our samples, 25 different ceramide compositions have been identified from 8 precursor (Z0) ions. Comparison of the two ionization modes (LSI and ESI) highlights the fact that molecular mass distributions obtained by LSI-MS, especially the presence of unsaturated species, have to be interpreted carefully. For the first time a direct high-performance liquid chromatography (HPLC)/ESI-MS method was used for characterization of complex mixtures of peracetylated GSLs. The results demonstrate that HPLC/ESI-MS is able to analyze mono- and diglycosylated GSLs, and other kinds of glycolipids that are actually present in the sample. PMID- 15536636 TI - A sensitive, rapid and specific determination of midazolam in human plasma and saliva by liquid chromatography/electrospray mass spectrometry. AB - A rapid, sensitive and selective liquid chromatography/electrospray mass spectrometry (LC/ES-MS) method was developed for the quantitative determination of the anaesthetic benzodiazepine midazolam (MID) in human saliva and plasma from patients undergoing anesthesia procedures. Biological samples spiked with diazepam-d5, the internal standard, were extracted into diethyl ether. Compounds were separated on a Xterra RP18 column using a mobile phase of acetonitrile/formic acid 0.1% at a flow rate of 0.25 mL/min under a linear gradient. Column effluents were analyzed using MS with an ES source in the positive ionization mode. Calibration curves were linear in the concentration ranges of 1-250 and 0.2-25 ng/mL in plasma and saliva, respectively. The limits of detection were 0.5 ng/mL in plasma and 0.1 ng/mL in saliva, using a 0.5-mL sample volume. The recoveries of the spiked samples were above 65%. The method was applied to ten real samples from patients undergoing midazolam treatment. PMID- 15536637 TI - Variation of mitochondrial size during the cell cycle: A multiparameter flow cytometric and microscopic study. AB - BACKGROUND: Changes in mitochondrial structure and size are observed in response to alterations in cell physiology. Flow cytometry provides a useful tool to study these changes in intact cells. We have used flow cytometry and digital fluorescence microscopy to analyze the variations in mitochondrial size in relation to specific phases of the cell cycle. METHODS: Supravital staining of rat fibroblasts was done with Hoechst 33342 and rhodamine 123, and cells were analyzed in a dual-laser flow cytometer. Synchronized cells at various stages of the cell cycle were analyzed for changes in mitochondrial size. These cells were also examined by electron microscopy, digital fluorescence microscopy and computerized image analysis to compare the lengths of the mitochondria. RESULTS: By using fluorescence pulse width analysis, we observed two populations of mitochondria in intact cells. The percentage of cells with small and large mitochondria at specific stages of the cell cycle indicated that mitochondrial size increases during the cell cycle; early G1 phase cells had the smallest mitochondria and the mitotic phase cells had the largest mitochondria. These results were confirmed by microscopic analysis of cells. CONCLUSIONS: Flow cytometry can distinguish the relative mitochondrial size in intact cells, and in combination with digital microscopy it can be used to study mitochondrial variation during the cell cycle. PMID- 15536638 TI - Quantitative analysis by flow cytometry of interstitial cells of Cajal, pacemakers, and mediators of neurotransmission in the gastrointestinal tract. AB - BACKGROUND: Interstitial cells of Cajal (ICCs) are mesenchymal cells that play critical roles in gastrointestinal motility as electrical pacemakers and mediators of neuromuscular neurotransmission. Although depletions of ICCs have been implicated in several gastrointestinal motor disorders, quantification of these cells has been difficult due to their varied morphology, regionally changing network density, and overall scarcity. Our goal was to evaluate flow cytometry (FCM) for the enumeration of ICCs. METHODS: We identified murine ICCs in live gastrointestinal muscles or primary cell cultures grown in the presence or absence of stem cell factor (SCF)-expressing STO fibroblasts with fluorescent Kit (CD117) antibodies. Because this technique also labels resident macrophages nonspecifically, we identified the latter with additional fluorescent antibodies. Dispersed cells were analyzed by FCM. RESULTS: ICCs represented 1.63 +/- 0.17% of the total cell count in the distal stomach (n = 18 mice) and 5.85 +/- 0.84% in the proximal colon and 6.28 +/- 0.61% in the distal colon (n = 3 mice). In fundic muscles of W/WV mice (n = 5) that virtually lack ICCs, very few Kit+ cells were detected. FCM identified approximately 2.6- to 7.3-fold more Kit+ ICCs in small intestinal cell cultures grown on STO fibroblasts expressing membrane-bound SCF (n = 6) than in cultures stimulated with soluble SCF (n = 6). CONCLUSIONS: FCM is a sensitive and specific method for the unbiased quantification of ICCs. PMID- 15536639 TI - Lectin-based three-color flow cytometric approach for studying cell surface glycosylation changes that occur during apoptosis. AB - BACKGROUND: Changes in cell surface glycosylation that accompany apoptosis are thought to be involved in the recognition and removal of apoptotic cells by phagocytes, but in most instances these changes are ill defined. To improve our understanding of this phenomenon, we designed a trivariate flow cytometry procedure that allows direct comparison of cell surface glycosylation in apoptotic and viable cells. METHODS: The annexin V/propidium iodide assay has been adapted for cell surface glycosylation analysis by combining the use of these two reagents with biotinylated lectins, and this has been used to investigate camptothecin-induced apoptosis in U-937 cells. RESULTS: Although numerous lectins are potent inducers of apoptosis, we found that it is possible to determine lectin concentrations that produce interpretable data without inducing significant cytotoxicity even when using apoptogenic lectins. That apoptosis is associated with a marked decrease in cell surface sialylation was confirmed by using the sialic acid-specific lectins Maackia amurensis agglutinin and Sambucus nigra agglutinin. These observations were corroborated by lectin blotting analysis with the same lectins. CONCLUSIONS: Species- and cell-dependent altered glycosylation patterns are likely to be associated with different modes of apoptosis. The easy and versatile method described in this report should be useful for exploring this field. PMID- 15536640 TI - Lack of beneficial effects of clonidine in the treatment of premenstrual dysphoric disorder: results of a double-blind, randomized study. AB - OBJECTIVES: To test the effects of clonidine in comparison with active placebo on premenstrual symptoms, mood scores and norepinephrine (NE) concentration, in women with premenstrual dysphoric disorder (PMDD). METHODS: Twelve women with prospectively confirmed PMDD were randomly assigned to oral 0.3 mg/day clonidine, as an active treatment, or 10 mg/day loratadine, as an active placebo, for 2 months each using a double-blind, cross-over design. NE concentration, premenstrual symptom ratings and mood scales were measured on three occasions: at pretreatment, after clonidine treatment and after placebo treatment. All patients were free of current psychiatric co-morbidity and medication use. RESULTS: There were no significant differences between clonidine and placebo for mood scales or premenstrual symptom ratings, though clonidine significantly suppressed NE concentration and produced more side effects in comparison with placebo. CONCLUSION: Compared with an active placebo clonidine demonstrated no beneficial changes in mood and premenstrual symptoms in women with PMDD. PMID- 15536641 TI - Regulation of lung cancer cell growth and invasiveness by beta-TRCP. AB - Beta-transducin-repeat-containing protein (beta-TRCP) serves as a substrate recognition subunit of Skp1/Cullin/F-box (SCF)(beta-TRCP) E3 ligases, involved in regulation of several important signaling molecules. SCF(beta-TRCP) E3 ligases play a critical role in cell mitosis as well as in various signaling pathways. Here, we provide evidence to support that beta-TRCP negatively regulates cell growth and motility of lung cancer cells. With specific antibodies, we detect loss of beta-TRCP1 protein in several lung cancer cell lines. One cell line contains an inactivated mutation of the beta-TRCP1 gene. Loss of beta-TRCP1 protein is also found in subsets of lung cancer specimens. We observe that retrovirus-mediated stable expression of beta-TRCP1 in beta-TRCP1 negative cells inhibits cell growth in soft-agar and tumor formation in nude mice. Furthermore, expression of beta-TRCP1 alters cell motility, as indicated by morphological changes and a reduced level of active matrix metalloproteinase (MMP)11. Conversely, inactivation of beta-TRCP1 by specific siRNA accelerates cell invasion. Of the 10 known substrates of SCF(beta-TRCP) E3 ligases, the protein level of cell division cycle 25 (CDC25)A is clearly affected in these lung cancer cells. Cells treated with CDC25A inhibitors become less invasive. Thus, loss of beta-TRCP1 may promote both growth and cell motility of lung cancer cells, possibly through regulation of CDC25A and the MMP11 level. PMID- 15536642 TI - Selecting fluorochrome conjugates for maximum sensitivity. PMID- 15536643 TI - Adhesive and mechanical properties of hydrogels influence neurite extension. AB - Photopolymerizable polyethylene glycol (PEG) hydrogels conjugated with bioactive ligands were examined for their use as scaffolds in peripheral nerve regeneration applications. The bioactivity and mechanical properties of PEG hydrogels can be tailored through the integration of bioactive factors (adhesion ligands, proteolytic sites, growth factors) and the alteration of PEG concentrations, respectively. For peripheral nerve regeneration, it will be important to determine the type and concentration of the bioactive molecules required to improve neurite extension. In this study, cell adhesion ligands (RGDS, IKVAV, and YIGSR) were covalently attached to PEG hydrogels. Both the type and concentration of cell adhesion ligand used affected neurite extension. Extension from PC12 cells was greater on hydrogels with RGDS incorporated than IKVAV, and the optimal concentration for each ligand was different. Cells adhered to but did not extend neurites on hydrogels with YIGSR. Cells did not adhere to hydrogels containing RGES. Furthermore, different combinations of these ligands affected neurite extension to different degrees. The mechanical properties of the hydrogels also significantly affected neurite extension. PC12 cells grown on more flexible hydrogels exhibited the greatest degree of neurite extension. PEG hydrogels have thus been developed with varying biochemical and mechanical properties that may enhance nerve regeneration. PMID- 15536644 TI - Ultrastructure of mesoderm in embryos of Opisthopatus roseus (Onychophora, Peripatopsidae): revision of the "long germ band" hypothesis for Opisthopatus. AB - In previous studies, an unusual pattern of development which resembles the "long germ band" development of some insects has been described in the onychophoran Opisthopatus cinctipes. This pattern has been proposed to be a characteristic of the genus Opisthopatus. To test this assumption, the ultrastructure of embryos of O. roseus, the sister species of O. cinctipes, was examined. Two kinds of paired, segmentally arranged coelomic cavities were found in the embryos studied: 1) dorsolateral coelomic cavities lined by extremely thin epithelia, and 2) ventral coelomic cavities situated within the anlagen of ventrolateral body appendages. Only the dorsolateral coelomic cavities can be considered "somites," since they occur earlier during embryogenesis. This is in contrast with the previous view that suggested a ventral position of "somites" in O. cinctipes. In addition, an anterior-to-posterior gradient occurs in the development of O. roseus. Based on our findings, we reevaluated the previous data on O. cinctipes. From this survey, no evidence in support of a "long germ band" hypothesis in Opisthopatus was found. Instead, the embryogenesis in representatives of Opisthopatus is more similar to that in other onychophorans than expected. PMID- 15536645 TI - Skeletal development of Macrochelys temminckii (Reptilia: Testudines: Chelydridae). AB - Few descriptions of the development and sequence of chondrification and ossification of the entire skeleton of turtles exist, particularly compared to other groups of reptiles. In this study, the embryonic skeleton and its ontogenesis are described for the Alligator Snapping Turtle, Macrochelys temminckii (Chelydridae). Morphological descriptions utilize cleared and double stained embryonic specimens and form the basis of comparison of the ontogenesis of the skeleton between this species and its extant sister taxon, Chelydra serpentina. The embryonic chondrocranium, as well as the sequences of formation and ossification of the entire skeleton, are compared between these closely related species, and afford a unique opportunity to examine differences in their patterns of skeletal formation. In M. temminckii, the first elements to ossify (Stage 17) are associated with the dermatocranium and upper jaw, followed by elements of the palate, lower jaw, and long bones of the limbs. In both species the majority of endochondral braincase elements (prootic, opisthotic, supraoccipital, and exoccipital) ossify after the majority of dermal elements of the skull. The sequences of formation of the chondral primordia of the limb elements, as well as ossification of autopodial elements, are generally congruent between these species. PMID- 15536646 TI - Ultrastructure of testicular macrophages in aging mice. AB - Testicular macrophages of aging mice were studied by TEM. Testicular macrophages retained with Leydig cells the close morphological relationships observed in the adult young animals, but digitations were not found. Lipofuscin granules like those of the Leydig cells from aging mice were observed in the cytoplasm. These organelles were generally absent in the testicular macrophages of young adult mice. Testicular macrophages did not display phagocytosis of the lipofuscin granules. In addition, the latter were not found in the intercellular spaces. These observations indicated that lipofuscin granules were formed, at least in a great part, within testicular macrophages as a consequence of metabolic changes occurring with age. Fine lamellar organization was seen in the lipofuscin granules of both Leydig cells and testicular macrophages. Frequently, lipofuscin granules originated from secondary lysosomes containing lipidic vacuoles only. Together with accumulation of the lipofuscin granules, changes of testicular macrophage fine morphology were observed. Endoplasmic reticulum and Golgi apparatus became poorly developed, and coated vesicles were rarely found. Fewer mitochondria were encountered, but their ultrastructure was not altered. These results suggest that in testicular macrophages lipofuscin accumulation is associated with a functional involution. PMID- 15536647 TI - Morphological evolution of the lizard skull: a geometric morphometrics survey. AB - Patterns of diversity among lizard skulls were studied from a morphological, phylogenetic, and functional perspective. A sample of 1,030 lizard skulls from 441 species in 17 families was used to create a lizard skull morphospace. This morphospace was combined with a phylogeny of lizard families to summarize general trends in the evolution of the lizard skull. A basal morphological split between the Iguania and Scleroglossa was observed. Iguanians are characterized by a short, high skull, with large areas of attachment for the external adductor musculature, relative to their sister group. The families of the Iguania appear to possess more intrafamilial morphological diversity than families of the Scleroglossa, but rarefaction of the data reveals this to be an artifact caused by the greater number of species represented in Iguanian families. Iguanian families also appear more dissimilar to one another than families of the Scleroglossa. Permutation tests indicate that this pattern is real and not due to the smaller number of families in the Iguanidae. Parallel and convergent evolution is observed among lizards with similar diets: ant and termite specialists, carnivores, and herbivores. However, these patterns are superimposed over the more general phylogenetic pattern of lizard skull diversity. This study has three central conclusions. Different clades of lizards show different patterns of disparity and divergence in patterns of morphospace occupation. Phylogeny imposes a primary signal upon which a secondary ecological signal is imprinted. Evolutionary patterns in skull metrics, taken with functional landmarks, allow testing of trends and the development of new hypotheses concerning both shape and biomechanics. PMID- 15536648 TI - Anatomy and histochemistry of spread-wing posture in birds. 3. Immunohistochemistry of flight muscles and the "shoulder lock" in albatrosses. AB - As a postural behavior, gliding and soaring flight in birds requires less energy than flapping flight. Slow tonic and slow twitch muscle fibers are specialized for sustained contraction with high fatigue resistance and are typically found in muscles associated with posture. Albatrosses are the elite of avian gliders; as such, we wanted to learn how their musculoskeletal system enables them to maintain spread-wing posture for prolonged gliding bouts. We used dissection and immunohistochemistry to evaluate muscle function for gliding flight in Laysan and Black-footed albatrosses. Albatrosses possess a locking mechanism at the shoulder composed of a tendinous sheet that extends from origin to insertion throughout the length of the deep layer of the pectoralis muscle. This fascial "strut" passively maintains horizontal wing orientation during gliding and soaring flight. A number of muscles, which likely facilitate gliding posture, are composed exclusively of slow fibers. These include Mm. coracobrachialis cranialis, extensor metacarpi radialis dorsalis, and deep pectoralis. In addition, a number of other muscles, including triceps scapularis, triceps humeralis, supracoracoideus, and extensor metacarpi radialis ventralis, were found to have populations of slow fibers. We believe that this extensive suite of uniformly slow muscles is associated with sustained gliding and is unique to birds that glide and soar for extended periods. These findings suggest that albatrosses utilize a combination of slow muscle fibers and a rigid limiting tendon for maintaining a prolonged, gliding posture. PMID- 15536649 TI - Heart inflow tract of the African lungfish Protopterus dolloi. AB - We report a morphologic study of the heart inflow tract of the African lungfish Protopterus dolloi. Attention was paid to the atrium, the sinus venosus, the pulmonary vein, and the atrioventricular (AV) plug, and to the relationships between all these structures. The atrium is divided caudally into two lobes, has a common part above the sinus venosus, and appears attached to the dorsal wall of the ventricle and outflow tract through connective tissue covered by the visceral pericardium. The pulmonary vein enters the sinus venosus and runs longitudinally toward the AV plug. Then it fuses with the pulmonalis fold and disappears as an anatomic entity. However, the oxygenated blood is directly conveyed into the left atrium by the formation of a pulmonary channel. This channel is formed cranially by the pulmonalis fold, ventrally by the AV plug, and caudally and dorsally by the atrial wall. The pulmonalis fold appears as a wide membranous fold which arises from the left side of the AV plug and extends dorsally to form the roof of the pulmonary channel. The pulmonalis fold also forms the right side of the pulmonary channel and sequesters the upper left corner of the sinus venosus from the main circulatory return. The AV plug is a large structure, firmly attached to the ventricular septum, which contains a hyaline cartilaginous core surrounded by connective tissue. The atrium is partially divided into two chambers by the presence of numerous pectinate muscles extended between the dorsal wall of the atrium and the roof of the pulmonary channel. Thus, partial atrial division is both internal and external, precluding the more complete division seen in amphibians. The present report, our own unpublished observations on other Protopterus, and a survey of the literature indicate that not only the Protopterus, but also other lungfish share many morphologic traits. PMID- 15536650 TI - Wear particles and surface topographies are modulators of osteoclastogenesis in vitro. AB - Prosthetic and osteosynthetic implants from metal alloys will be indispensable in orthopedic surgery, as long as tissue engineering and biodegradable bone substitutes do not lead to products that will be applied in clinical routine for the repair of bone, cartilage, and joint defects. Therefore, the elucidation of the interactions between the periprosthetic tissues and the implant remains of clinical relevance and several factors are known to affect the longevity of implants. Within this study, the effects of metal particles and surface topography on the recruitment of osteoclasts was investigated in vitro in a coculture of osteoblasts and bone marrow cells. The cells were grown in the presence of particles of different sizes and chemical composition or on metal discs with polished or sandblasted surfaces, respectively. At the end of the culture, newly formed osteoclasts were counted. Osteoclastogenesis was reduced when particles were added directly to the coculture. The effect depended on the size of the particles, small particles exerting stronger effects than larger ones. The chemical composition of the particles, however, did not affect the development of osteoclasts. In cocultures grown on sandblasted surfaces, osteoclasts developed at higher rates than they did in cultures on polished surfaces. The data demonstrate that wear particles and implant surfaces affect osteoclastogenesis and thus may be involved in the induction of local bone resorption and the formation of osteolytic lesions, leading eventually to the loosening of orthopedic implants. PMID- 15536651 TI - Dorsal fin in the white shark, Carcharodon carcharias: a dynamic stabilizer for fast swimming. AB - Transverse sections of the skin in the dorsal fin of the white shark, Carcharodon carcharias, tiger shark, Galeocerdo cuvier, and spotted raggedtooth shark, Carcharias taurus, show large numbers of dermal fiber bundles, which extend from the body into the fin. The bundles are tightly grouped together in staggered formation (not arranged in a straight line or in rows). This arrangement of dermal fibers gives tensile strength without impeding fiber movement. Tangential sections indicate that the fibers in all three species are strained and lie at angles in excess of 60 degrees . Of the three species investigated the dermal fibers in C. carcharias are the most densely concentrated and extend furthest distally along the dorsal fin. The overall results indicate that the dorsal fin of C. carcharias functions as a dynamic stabilizer and that the dermal fibers are crucial to this role. The fibers work like riggings that stabilize a ship's mast. During fast swimming, when the problems of yaw and roll are greatest, hydrostatic pressure within the shark increases and the fibers around the body, including in the dorsal fin, become taut, thereby stiffening the fin. During slow swimming and feeding the hydrostatic pressure is reduced, the fibers are slackened, and the muscles are able to exert greater bending forces on the fin via the radials and ceratotrichia. In C. carcharias there is a trade-off for greater stiffness of the dorsal fin against flexibility. PMID- 15536652 TI - Surgical treatment of patent ductus arteriosus: a new historical perspective. AB - The conventional surgical history of ligation of a patent ductus arteriosus (PDA) dates from August 26, 1938, when Robert E. Gross of Boston, Massachusetts, successfully ligated a PDA. It is largely unknown that in the same year and before Gross, Emil Karl Frey, a Surgeon at the Medizinische Akademie in Dusseldorf, Germany, already ligated a PDA successfully. Assuming that he would soon perform more ligations, Frey did not publish his findings, and this historic ductal operation escaped attention. PMID- 15536653 TI - Expression and localization of steroid receptors in human nasal mucosa. AB - OBJECTIVE: To investigate the expression of glucocorticoid receptor (GR), oestrogen receptor (ER), progesterone receptor (PR) and androgen receptor (AR) in nasal mucosa. MATERIAL AND METHODS: Human turbinates were obtained after turbinectomy from seven patients. The expression and localization of steroid receptors were examined using reverse transcriptase polymerase chain reaction (RT PCR) and immunohistochemistry. RESULTS: Using RT-PCR, GR and ER alpha mRNA were detected in all cases. In contrast, ER beta, PR and AR mRNA were found in five, four and six cases, respectively. Using immunohistochemistry, antibodies to GR showed the presence of GR within all cells of nasal mucosa, with the highest quantities of GR being localized in epithelial cells, submucosal glands and inflammatory leukocytes. Immunohistochemical analysis of sex steroid receptor revealed that anti-ER alpha antibody labelled mainly mast cells and anti-ER beta antibody labelled submucosal glands, and that no PR or AR expression was detected in any of the samples tested. CONCLUSIONS: The role of ER in mast cells and submucosal glands has not been well clarified. However, precise knowledge of the identity and distribution of sex steroid receptor should be of considerable interest in understanding the role of sex hormones in upper airway diseases such as allergic and non-allergic rhinitis. PMID- 15536654 TI - Influence of the hydrophobic and hydrophilic characteristics of sliding and slider surfaces on friction coefficient: in vivo human skin friction comparison. AB - INTRODUCTION: The objective of this study was to investigate whether hydrophilic/hydrophobic balance (Hi/Ho) of the skin surface strongly modifies the friction coefficient (mu). The Hi/Ho balance is determined using the relationship between the critical surface tension gammac (Zisman's principle:which delimits the wetting capacity) and the surface tension of water gammaH2O (water: reference element of Hi/Ho balance). METHOD: Critical surface tension gammac was determined (according to Zisman's principle) through the measurement of advancing contact angle theta of a series of ethanol/water dilutions. Friction coefficient depends on several parameters: types of probe motions (rotational vs. linear), surface roughness and physicochemical parameters of surfaces in contact). In this study, the wettability parameters for six surfaces (human skin forearm, Teflon, silicone impression material 'Silflo', vinyl polysiloxane impression material 'resin' steel and glass) were measured and their influences were compared to friction coefficient mu. RESULTS: This study shows that the higher hydrophobia tendency of the surfaces, the lower friction coefficient. The use of three sliding materials (Teflon, steel and glass) of different Hi/Ho balance confirms the importance of these physicochemical parameters in mu. For example, Teflon with high hydrophobia has a low mu. Friction coefficient increased when hydrophobia of sliding and slider surfaces decreased. CONCLUSION: Friction coefficient value depends on the type of slider surface and its physicochemical properties. In vivo,the friction coefficient may quantify the influence of lubrificant/emolients/moisturizers. For example, the friction coefficient of hydrated skin (through the action of moisturizing products) is higher than the friction coefficient of dry skin. The relationship between the friction coefficient and the Hi/Ho balance can be reversed in the presence of water and sebum on forehead, for example. PMID- 15536655 TI - Objective measurements of radiotherapy-induced erythema. AB - BACKGROUND/AIMS: The development of acute radiation erythema is a common phenomenon among patients under-going radiotherapy treatment. Because of the absence of reliable objective classification methods, the degree of skin reaction can at present mainly be judged subjectively in the clinic. This has motivated the present preliminary study,concerning the first steps in the development of an objective method for skin reaction classification. METHODS: Three non-invasive techniques were used:near-infrared (NIR) spectroscopy, laser Doppler perfusion imaging and digital photography. The NIR spectra were analysed with principal component analysis (PCA), and the results from the other two with traditional univariate methods. Measurements were made on breast cancer patients who had been exposed to different irradiation doses. A total of 28 breast cancer patients participated one to three times each; 12 were treated with photons at 4 or 6MeV and 16 were treated with high-energy electrons between 10 and 20 MeV to a maximum dose of 50 Gy. RESULTS: PCA of NIR spectra shows that information on radiation dose lies mainly in the first principal component. It is observed that the higher the dose the higher the score value. The results from the laser Doppler measurements show that in 79% of the cases the perfusion increases significantly with radiation dose. Analysis of the digital photography shows that a proposed skin redness index(SRI), increases with a higher radiation dose. However,the increase in most cases is not significant. By combining all data, correlation to radiation doses was seen for 74% of the patients who participated more than once. CONCLUSION: All three non-invasive methods correlate with the radiation dose but to various degrees. NIR spectroscopy, laser Doppler and a combination of the three techniques are the most promising methods for characterising erythema. PMID- 15536656 TI - Ptosis of submental skin: objective measurements and effect of age. AB - BACKGROUND/PURPOSE: Sagging of the face skin is a visible and unattractive characteristic of skin ageing. This phenomenon, due to both gravitational force and age-related alterations in the mechanical properties of skin, has never been quantitatively studied. The aim of this paper was, first, to define a method and a precise procedure allowing the objective measurement of this phenomenon and, second,to study how it is related to age. METHODS: We used an electronically controlled bed that allows the comparison of submental skin profiles in both the reclined and the upright positions. A parameter that characterises the sagging amplitude for an individual was defined and the reproducibility of measurement of this parameter was verified. RESULTS: Results, obtained from 66 female volunteers, demonstrated a statistically significant positive correlation between this new parameter and age, and also between this new parameter and the elastic recovery parameter of skin, as measured by torquemeter. CONCLUSION: Age-related changes of the skin's mechanical properties and of the various underlying skin tissues would explain this phenomenon. This new method and procedure should allow objective evaluation of the efficacy of any treatments (cosmetic, medical or surgical) proposed for rendering the appearance of people younger. PMID- 15536657 TI - Exploring the influence of skin perfusion on transepidermal water loss. AB - BACKGROUND: Eventual relationships between the vascular function and transepidermal water loss (TEWL), in vivo,have not been entirely explored. By promoting local perfusion alterations through a well-known challenge test, the'tourniquet-cuff occlusion' manoeuvre, the present study searches for other dynamical factors influencing the cutaneous barrier, further exploring the applicability of these flow-related variables in dermatological research. METHODS: By applying the tourniquet-cuff manoeuvre to a group of healthy volunteers (n= 20), transcutaneous (tc)gases (pO2-pCO2), LDF (laser doppler flowmetry) and TEWL were considered as representing the dynamical aspects under study and measured non-invasively. RESULTS AND CONCLUSION: An haemodynamical relationship between tcpO2 and LDF in the post-occlusive period was clearly identified, defining the autoregulatory index as a numerical descriptor of the local metabolic-flow adjustment under stress. TEWL was also significantly affected by the manoeuvre, especially during the post-occlusive period,although no significant relationships between TEWL and other tc variables could be found. The present findings seem to suggest that, under the present experimental conditions, local haemodynamics may also influence TEWL measurements and the skin barrier. PMID- 15536658 TI - Towards objective evaluation of the skin aspect: principles and instrumentation. AB - BACKGROUND/AIMS: The aim of this paper is to propose an objective procedure for the evaluation of the skin aspect. This work is based on light scattering phenomena and in-vivo measurements of the skin profile and of the skin Bi directional Reflection Distribution Function (BRDF). METHODS: We consider the basic mechanisms of light reflection and scattering by a surface to extract the key sur-face-parameters of aspect. It appears that the surface relief is determinant for the local surface orientation while the light scattering properties of the skin are responsible for the visibility of that skin relief. The first part presents the principles of skin-image formation on the retina and de-scribes the actual roles of the BRDF and of the skin relief. The second part is dedicated to experimental measurements on the skin in vivo. The 3D relief is reconstructed by a fringe projection method and numerical processing. The angular distribution of light reflected by the skin is also measured by means of a different set-up. RESULTS: The knowledge of both the skin relief obtained by 3D optical profilometry and the skin scattering behavior measured for regularly distributed set angle values, allows an objective aspect evaluation as well as a prediction of the skin images that an observer will receive. CONCLUSION: This paper presents both the principles and instrumentation elements suitable for the setting up of an objective evaluation procedure of the skin aspect. PMID- 15536659 TI - Experiments on peeling adhesive tapes from human forearms. AB - BACKGROUND/PURPOSE: Peeling of adhesive tape from hu-man skin can cause pain and trauma. The effects of the peel angle, peel rate, dwell time, subject, and order of testing were examined. METHODS: Adhesive tape was peeled from the volar forearms of four human subjects. The tape was 25.4-mm-wide Durapore by 3M. The rates ranged from 100 to 10,000 mm/min, the peel angle from 90 degrees to 180 degrees, and the dwell time from one to 15 min. The tensile testing machines'recorded the peel force and the displacement of the end of the tape. RESULTS: A range of maximum and average peel force values was observed. The peel force tended to be minimum for a peel angle around 150 degrees in most cases. As the peel rate was increased, the peel force generally tended to increases lightly, but the relationship was erratic. As the dwell time increased, the peel force increased and then settled down. Of the subjects tested, the skin of the oldest subject experienced the most displacement and highest peel forces(the maximum force over all tests was 3.6 N). The peel force tended to increase with repeated peeling from the same skin site. Compared with peeling from a rigid substrate such as steel, the peel force from skin was lower. CONCLUSION: The results improve understanding of the factors affecting the peel force, which has been correlated previously with trauma caused by peeling adhesive tape from skin. PMID- 15536660 TI - Coping with mild inflammatory catamenial acne: a clinical and bioinstrumental split-face assessment. AB - BACKGROUND: Acne is a multifactorial disease exhibiting distinct clinical presentations. Among them, the catamenial type is a matter of concern for young women. Some oral contraceptives may help without, however, clearing the skin condition. AIM: The present open study aimed at evaluating the effect of overnight applications of a paste made of petrolatum,15% zinc oxide and 0.25% miconazole nitrate. METHOD: The split-face trial was conducted in 35 women. A non medicated cream was used as control. Clinical evaluations and biometrological assessments on cyanoacrylate follicular biopsies were performed monthly for 3 months. Comedometry and the density in autofluorescent follicular casts were used as analytical parameters. In addition, the five most severe cases at inclusion were tested at the completion of the study for follicular bacterial viability using dual flow cytometry. RESULTS: Compared with baseline and to the control hemi-face, the medicated paste brought significant improvement of acne. The number of papules and their redness were reduced beginning with the first treatment phase. A reduction in the follicular fluorescence was yielded beginning with the second treatment phase. The ratios between injured and dead bacteria, on the one hand, and live bacteria, on the other hand were significantly increased at completion of the study. CONCLUSION: A miconazole paste applied for 1 week at the end of the ovarian cycle has a beneficial effect on catamenial acne. PMID- 15536661 TI - Assessment of spray application of Saint GERVAIS water effects on skin wettability by contact angle measurement comparison with bidistilled water. AB - BACKGROUND: The skin is responsible for protecting the body from physical, chemical and microbial injuries. The stratum corneum is the top layer of the epidermis and it plays a key role in helping to contain moisture. When the skin becomes damaged, its ability to perform these functions is compromised. Dry skin is a common form of skin damage. Contact angle 0 between a surface and water is a good indicator of hydrophobic or hydrophilic tendency of surfaces. OBJECTIVE: To quantify the influence of spray application of an isotonic mineral water (Saint GERVAIS water) on the hydrophobic tendency of dry skin and to compare spreading of both types of water on the skin through 0 measurement of each water drop on the skin before and after mineral spray application. MATERIALS AND METHODS: Contact angles theta between skin surface and the two types of water (bidistilled and mineral water) were measured before and after application of mineral water by a system allowing the measurement of theta in vivo and in vitro. The cutaneous hydration, the temperature and pH were also measured before and after mineral water application. RESULTS: As for the application of mineral water, the cutaneous pH significantly increased so the strong skin acidity decreased. The cutaneous temperature was modified (a decrease of 2 degrees C was noted) and the cutaneous hydration was also increased (+47AU). The hydro-phobic tendency of the dry skin tested in this study was strongly decreased after mineral water application. This parameter was illustrated by a decrease of contact angle theta value. CONCLUSION: Saint GERVAIS water application allowed a regulation of skin pH, a fresh sensation on the skin and an increase in the cutaneous hydration. Moreover, the contact angle measurement with bidistilled water illustrated the increased wettability of the skin. PMID- 15536662 TI - High-yield single-photon source using gated spontaneous parametric downconversion. AB - The construction of a single-photon source by use of gated parametric fluorescence is reported with the measurement results of the photon number distribution. A beamlike twin-photon method is used in order to achieve high collection efficiency. The estimated probability P(1) to find a single photon in a collimated output pulse is 26.5% at a repetition rate of 10 kHz when the effective quantum efficiency of 27.4% in the detection setup is compensated. PMID- 15536663 TI - Diffraction theory in TM polarization: application of the fast Fourier factorization method to cylindrical devices with arbitrary cross section. AB - The diffraction of an electromagnetic wave by a cylindrical object with arbitrary cross section is studied by taking advantage of recent progress in grating theories. The fast Fourier factorization method previously developed in Cartesian coordinates is extended to cylindrical coordinates thanks to the periodicity of both the diffracting object and the incident wave with respect to the polar angle theta. Thus Maxwell equations in a truncated Fourier space are derived and separated in TE and TM polarization cases. The new set of equations for TM polarization is resolved numerically with the S-matrix propagation algorithm. Examples of elliptic cross sections and cross sections including couples of nonconcentric circles show fast convergence of the results, for both dielectric and metallic materials, as well as good agreement with previous published results. Thus the method is suitable for an extension to conical (out-of-plane) diffraction, which will allow studying mode propagation along microstructured fibers. PMID- 15536664 TI - [Cardiac pacemaker in the catheter laboratory?]. PMID- 15536665 TI - Prevalence of hypertrophic cardiomyopathy in China. PMID- 15536666 TI - Physical exposures in work commonly done by women. AB - The North American work force is still highly sex-segregated, with most members of each sex in jobs composed primarily of workers of the same sex. This division is accentuated when jobs involve physical demands. Women have traditionally been assigned to tasks whose physical demands are considered to be light. Nevertheless, these tasks can have biological effects, sometimes serious. Phenomena related to physical demands of women's work can be considered in three categories: (a) musculoskeletal and cardiovascular demands of tasks often assigned to women in factories and service work; (b) sex- and gender-specific effects of toxic substances found in the workplace; and (c) interactions between work and the domestic responsibilities of many women. These phenomena are described, using examples recently gathered from workplaces. Effects of biological sex are distinguished, as far as possible, from effects of gender (social roles). PMID- 15536667 TI - Physical activity and atherosclerosis: which animal model? AB - Atherosclerosis is a progressive disease that is the most important single contributor to human cardiovascular morbidity and mortality. Epidemiologic studies show that physical activity, or routine exercise, reduces the risk of developing cardiovascular disease. The mechanisms through which exercise may function in primary or secondary prevention of atherosclerosis remain largely to be established. Most studies in humans are performed after the onset of clinical signs when disease is well advanced and the prescription of exercise is based on empirical evidence of benefit in secondary prevention. Animal models per-mit the study of the initiation and progression of preclinical stages of atherosclerosis. In order to provide information relevant to treatment and prevention, these models should mimic human disease and interactions of physical activity with disease processes as closely as possible. The purpose of this review is to compare animal models of atherosclerosis and to summarize the available data in those models in regard to the effects of exercise. PMID- 15536669 TI - Stem cells: current usage and future potentials. PMID- 15536668 TI - Fluorescence and MALDI on a single chip. PMID- 15536670 TI - Economic cost and epidemiological characteristics of fibromyalgia. PMID- 15536671 TI - The treatment of enthesitis in psoriatic arthritis. PMID- 15536672 TI - Casemix in Singapore - 5 years on. PMID- 15536673 TI - Flap prefabrication - the bridge between conventional flaps and tissue-engineered flaps. AB - Flap prefabrication is one of the most exciting areas in Plastic Surgery because of its bridging role between conventional reconstructive surgery and tissue engineering. Using this technique,tissues such as bone, cartilage, skin and muscle can be preassembled to form precise composites that will fit any defect. In pre-lamination, for example, an ear may be created by burying cartilage underneath forearm skin and later harvested as a skin-cartilage composite free flap to replace the missing part. Vascular induction is yet another means of customising flaps where new blood supply is introduced to create transplantable tissue. For example, bone chips wrapped in avascular carrier such as muscle can become vascularised grafts. Our experiment describes jejunal prefabrication in a rat model using the same technique. Intestinal segments wrapped in muscle flaps become independent of their mesenteric blood supply by "parasitising" on the muscle's blood supply. This idea arose from our initial observations that intestinal segments transferred to the neck to reconstruct the oesophagus could survive accidental disruption of the pedicle if sufficient time had elapsed. Clearly, the bowel had picked up new blood supply from its bed. Subsequently, jejunal prefabrication was used to reconstruct the oesophagus in a patient in whom there were no recipient vessels for free jejunal transfer. The pedicled latissimus dorsi muscle flap was used as a carrier for the jejunum. Another application of this idea could be in the area of allogeneic trachea or pancreatic transplantation, since present-day techniques have yet to overcome problems such as insufficient vascularity and unpredictable transplant survival. Future applications incorporating biomaterials and cultured cells will usher in the era of tissue-engineered flaps. PMID- 15536674 TI - The role of public health and occupational physicians in the new millennium. PMID- 15536675 TI - Reflections of a septuagenarian - radiology: past, present and future. PMID- 15536676 TI - Katharina Dorothea Dalton. PMID- 15536677 TI - Pituitary in psychosis. PMID- 15536678 TI - CBT for psychosis. PMID- 15536679 TI - The trouble of transporting radioactive materials. PMID- 15536680 TI - New UK guidelines for use of immunosuppressive agents. PMID- 15536681 TI - Shine the magnetic resonance light on neurosurgery. PMID- 15536682 TI - WHO puts cancer on global health agenda. PMID- 15536683 TI - Progress in cancer fight for developing world. PMID- 15536684 TI - New type of effective antifungal treatment identified. PMID- 15536685 TI - Cancer survivors receive less care for other medical conditions. PMID- 15536686 TI - Second-generation antisense drug for prostate cancer. PMID- 15536687 TI - ERBB2 kinase mutations in lung-cancer tumours. PMID- 15536688 TI - Rash correlates with tumour response after cetuximab. PMID- 15536689 TI - Cottonseed-oil byproduct as anticancer radiation sensitiser. PMID- 15536690 TI - Siberia's shrouded legacy. AB - Victoria Dvornichenko is director of Irkutsk Regional Oncology Centre and an oncologist for the Irkutsk region in eastern Siberia, Russian Federation. She has worked as an oncologist for 32 years and was a senior consultant during the break up of the Soviet Union and the transition from central planning to a market economy. This transition has been more painful in Russia than in the central European countries that joined the European Union in May, 2004, and the 1990s in Russia were associated with a severe economic decline. But healthcare was given a high priority under the Soviet system, and this priority has remained throughout the transition. Investment in cancer care and research is still substantially high. PMID- 15536691 TI - You, me, and cancer: BBC three cancer season. PMID- 15536692 TI - Graves' disease with unilateral radioisotope uptake. PMID- 15536693 TI - [Uncomplicated pregnancy and birth of a normal child in thyroid cancer after high dose 68 Gbq iodine 131]. PMID- 15536694 TI - Staining sections with Giemsa. PMID- 15536695 TI - Donkey: hero or villain of the parasite world? Past, present and future. AB - This collection of articles provides an account of five presentations delivered at the 19th International Conference of the World Association for the Advancement of Veterinary Parasitology(WAAVP) (held in New Orleans, Louisiana, USA, from 10 to 14 August 2003) in a symposium session on Donkey parasites, organised by Andrew Trawford and Catherine Morriss and moderated by Dr. Elisabeth Svendsen. The focus was on the role of the donkey in the advancement of parasitology, diagnosis and identification, methods of control and anthelmintic resistance. PMID- 15536696 TI - [Myocarditis. A misdiagnosed common illness?]. PMID- 15536697 TI - [Cardiovascular risk: a class effect of coxibs? How to manage patients taking Vioxx?]. PMID- 15536698 TI - [Controlling symptoms as early as possible. Palliative medicine is not the final anointment]. PMID- 15536699 TI - [Acute manifestations due to allergen contact. Apples and nuts can be poisons in neurodermatitis]. PMID- 15536700 TI - [2004 Nobel Prize for Medicine. A good sense of smell for the entire body (interview by Dr. T. Meissner)]. PMID- 15536701 TI - [Plastic surgery is more than a means to achieve a perfect figure]. PMID- 15536702 TI - [The role of plastic surgery in remodeling the body image]. AB - On the basis of comprehensive information on the realistic possibilities and outcomes of esthetic corrective surgery, such operations can contribute to an effective and lasting improvement in the patient's quality of life and self esteem. However, the esthetic surgeon's scalpel must never be considered an alternative to necessary psychotherapy. Exaggerated expectations, obsessions, and an absent sense of reality coupled with the projection of psychological and social problems on a certain part of the body must be interpreted as dysmorphophobia with a disordered personality structure. For a reputable plastic surgeon they must be considered a contraindication to esthetic surgery. Against a background of constantly changing social conditions and attitudes, too, surgical body sculpting should no longer be uncritically accepted as a legitimate field for esthetic surgery. Currently, the question as to where the lines are to be drawn for the performance of body-modeling surgical interventions remains unanswered. PMID- 15536703 TI - [Breast-reduction surgery--a long-term survey of indications and outcomes]. AB - Between 1986 and 2003, breast-reduction surgery was performed in a total of 814 women. The indication was established on the basis of physical complaints, chronic back pain, stiff neck or recurrent intertrigo in the foldbeneath the breasts. A proportion of the patients were interviewed postoperatively using a questionnaire, to determine the impact of the operation on their quality of life. 91% of those surveyed reported a postoperative improvement in the perception of their own body, and 80% were satisfied with the reduced size of their breasts. In conclusion, in the hands of an experienced breast surgeon, breast-reduction surgery for the proper indication results in a reliable and safe diminishment in breast size and tightening of slack tissue, leading to a significant enhancement in the patient's quality of life. PMID- 15536704 TI - [Possiblities for breast reconstruction following cancer surgery]. AB - Cancer of the breast is the most common malignant disease of women, and currently affects approximately 10% of all women. In the large majority of cases, mammary carcinoma can be treated without having to sacrifice the breast. In some 30% of the cases, however, amputation continues to be needed, in particular when a relatively large tumor presents, or when its removal with an adequate safety margin in a relatively small breast would result in appreciable deformation. Reconstruction of the breast is often an important psychological factor, since, as a visible organ, the breast is a determining feature of a woman's physical appearance and her effect on the opposite sex. The availability of various procedures and modern techniques of breast reconstruction make it possible to comply with the individual wishes of the woman concerned. PMID- 15536705 TI - [Diagnosis and treatment of panic disorder]. AB - Panic disorder is defined as, recurrent, unexpected panic attacks. Panic attack means a period of intensive fear or discomfort, accompanied by a range of physical or psychological symptoms. Subsequently, anticipatory anxiety and avoidance behavior often develop. Panic disorder may or may not be accompanied by agoraphobia. Panic disorder can be treated both by psychotherapeutic and pharmacological measures. An ideal approach is combination therapy in the sense of a multimodal concept; with regard to cognitive behavioral therapy in particular, its effectiveness in the treatment of panic disorder is well documented. For pharmacotherapy, antidepressive agents, in particular selective serotonin reuptake inhibitors, are the drugs of first choice. Benzodiazepines should be given for only a few weeks until the antipanic effect of the antidepressants kicks in. With appropriate treatment, the prognosis is favorable. PMID- 15536706 TI - [Diabetes and Nephropathy: ACE inhibitor for type 1, AT1 blocker for type 2]. PMID- 15536707 TI - [Patient dissatisfied with treatment outcome. Increasingly physicians are blamed]. PMID- 15536708 TI - [Family physicians group strives for autonomy. Family physicians want to manage reimbursement independently]. PMID- 15536709 TI - [Long-term oral anticoagulation. Evidence favors self management]. PMID- 15536710 TI - [Calculate stroke risk for the patient! Concrete numbers are better motivation for patients]. PMID- 15536711 TI - [Show tests of stroke risk: "There can not be enough patient education"]. PMID- 15536712 TI - [Diagnosis at a glance? Nodule of the ear]. PMID- 15536713 TI - [An unappealing thing: half miniature duffel bag, half inflated mutton tube]. PMID- 15536714 TI - Applicability of Rosenstock-Hochbaum health behaviour model to prevention of periodontal disease in Enugu students. AB - From a survey of seven hundred students whose ages ranged from fourteen to twenty one, three hundred were selected on the basis of regular brushing and having seen a dentist for 'routine' check up. Their responses were analysed to see if their knowledge and behaviour conform to the Rosenstock-Hochbaum health belief model. It was found that they had little knowledge of the causes of dental diseases. Nevertheless they were those that brushed regularly and claimed to have gone for routine check up. This then casts some doubt about the applicability of Rosenstock-Hochbaum model of health behaviour to prevention of death diseases. PMID- 15536715 TI - Association between temporo mandibular disorders and oral parafunctions in Saudi children. AB - The purpose of the present investigation was to determine the relationship between oral parafunctions and the development of temporomandibualr disorders (TMD). A group of 1,976 children aged 3-15 years old, were randomly selected and underwent an examination that consisted of palpation and assessment of the temporomandibular joints and associated muscles for tenderness and joint sounds. Maximum vertical opening and deviation during jaw opening were also recorded. The parents were requested to complete a questionnaire contained questions regarding symptoms of TMD and history of oral parafunctions. Results showed significant correlations between cheek biting, nail biting, bruxism, thumb sucking and most of TMD signs and symptoms. The results of the study support the concept that oral parafunctions have a significant role in the aetiology of TMD and should be tackled by the paediatric dentists to minimise their effects on the stomatological system. PMID- 15536716 TI - [Quantification of initial malocclusion according to the mode of breathing in black African children]. AB - The relations between the mode of breathing and the development of the malocclusions were the subject of many studies causing polemic sometimes (2, 3, 7, 9, 20). In fact the impact of the mode of breathing on occlusion is not clarified yet. The goal of this study is to quantify the dental characteristics, which constitute the malocclusion according to the mode of breathing. 100 African melanoderme children old from 6 to 15 years were subjected to a rhinologic evaluation based on the nostril reflex of GUDIN and the test of ROSENTHAL (12). Of this examination these children were left again in a group of 50 nasal respirators and in another group of 50 mouth breathers. Each child underwent a radiographic examination which was used to make a cephalometric analysis and a meeting of catch of dental prints. The statistical analysis of the data recorded on the dental casts and the layouts cephalometric (test t of student) indicate that the mode of breathing is not associated standard initial malocclusion. But, when the facial divergence, which is characteristic of mouth breathing increases, the initial malocclusion becomes significant. PMID- 15536717 TI - Oral health knowledge and sources of information among male Saudi school children. AB - The purpose of present study was to determine the oral health knowledge and sources of information in male Saudi school children. The required information was collected through a especially designed questionnaire. A total of 130 children completed the questionnaire with the mean age of 13.3 (SD 1.9) years. There was no significant difference in oral health knowledge or sources of information in relation to age and educational level. Less than half (44.6%) of the children actually had heard about fluoride, one-third (34.6%) correctly identified the action of fluoride as preventing tooth decay. Almost all (97.2%) the children thought that sweets (chocolates/candies) could cause tooth decay. However, a large number of children were not aware of cariogenic potential of soft drinks (31.5%). More than half (53.1%) of the children reported that their dentist taught them how to brush properly. However, 11.5% children were not taught by any one about proper tooth-brushing. A large number (40.0%) of children thought that one must visit the dentist only in case of pain in the teeth. Dentists were the most popular (61.5%) source of oral health information. It can be concluded that the children need further oral health education in areas of caries prevention, and there was a need to utilise parents, schoolteachers and media to enhance their oral health knowledge. PMID- 15536718 TI - [Hypersialorrhea in mental retardation: interest of salivary denervation]. AB - Hypersialorrhea with mental retardation is a handicapping clinical situation with a greater psychoaffective impact and a risk of dyspneic attack by a pharyngeal salivary stasis and a disappointing treatment. We report an observation about a 22 years old child; as for as this child is concerned, the treatment has been a salivary denervation. This method is simple, efficient and without risk. PMID- 15536719 TI - Extrinsic tooth discolouration in 11-16 year-old Nigerian children. AB - This study was done to ascertain the prevalence and aetiology of extrinsic stains among 11-16 year-old school children in Surulere, which is one of the Local Government Areas (LGAs) in Lagos, Nigeria. 600 school children were randomly selected from 6 schools, 3 primary and 3 secondary, in the LGA. About 100 children were examined from each age category of 11 to 16 years for extrinsic stains, and information on their dietary and oral habits was obtained. A 64.0% prevalence of extrinsic stains was found among the study population (boys =64.5%, girls = 63.4%). The colour of the extrinsic stains ranged from yellow, brown, green, orange, black to red and were mainly due to poor oral hygiene. The extrinsic stains were derived mainly from food consumption and the action of chromogenic bacteria. The high level of extrinsic stains seen in this study is also a reflection of the low utilisation of dental services and subsequently of professional oral prophylaxis among the subjects as 91.5% had never visited a dentist and 94.8% had never had a professional oral prophylaxis. The prevalence of stains seen in the study population is higher than would be expected in adolescents who should be rather self-conscious at that age, the older adolescents even having a significantly higher prevalence of stains than younger adolescents (p < 0.05). No statistical difference in extrinsic stains prevalence was found among the sexes (p = 0.7) or among social classes (p> 0.05). The establishment of a school oral health care programme in the Local Government Area is advocated, as this may be the only source of oral care for these children. Dental health education and promotion services should be the main thrust of such services, which may be in form of fixed or mobile dental clinics. PMID- 15536720 TI - Prevalence of intrinsic tooth discolouration among 11-16 year-old Nigerians. AB - 600 children aged 12 to 16 years were examined in the Surulere Local Government Area of Lagos State, Nigeria to determine the prevalence and types of intrinsic tooth discoloration. 306 boys and 294 girls were selected from six schools (3 primary and 3 secondary schools) by stratified random sampling. The teeth of the children were examined in a wet state for intrinsic stains, however, no attempt was made at aetiologic diagnosis of enamel hypoplasia and enamel opacities, diagnosis were based on clinical presentations. A 37.7% (226) prevalence of intrinsic tooth discolouration was recorded among the subjects. 36.0% (110) of the boys and 39.5% (116) of the girls had intrinsic tooth discolourations. The differences in prevalence of intrinsic tooth discolouration among the sexes and among the social classes were not statistically significant (p = 0.375 and p = 0.272). There was, however, a statistical difference in prevalence of intrinsic tooth discolouration among the age categories (p = 0.0013), with the older adolescents showing a higher prevalence of stains. This is linked to the greater possibility of presenting with acquired intrinsic tooth discolouration as one gets older. The commonest types of intrinsic tooth discoloration were enamel opacities (23.0%), tetracycline staining (9.2%) and enamel hypoplasia (7.3%). Other intrinsic tooth discolourations were those due to dental caries (4.3%), pulp necrosis/ haemorrhage (0.8%), toothwear (0.3%), and discoloured restorations (0.3%). A very low level of utilisation of dental services was noted, with 91.5% (549) of the subjects reporting that they had never visited a dentist. The common intrinsic tooth discolorations seen in the study are those usually associated with childhood malnutrition, preventable childhood diseases and the irrational use of drugs (tetracyclines). There is a need to intensify child immunization efforts in the community as well as health education and promotion activities geared toward proper nutrition and the rational use of drugs. The establishment of school meals in primary and secondary schools in the LGA will go a long way in improving nutrition and prevent the tooth discolouration types associated with poor nutrition. Dental services may be made more available, accessible and affordable for the children by the establishment of a school oral healthcare programme. PMID- 15536721 TI - [The place of chemical products in oral hygiene for the prevention and treatment of periodontal disease]. AB - The mechanical elimination of the bacteria plaque is the basis of the prevention and the treatment of gingivitis and periodontitis. Chemicals products used in the control of supra and subgingival plaque show a great importance because of individual and professional difficulties which constitute an impediment to the effective elimination of the bacteria plaque. However, by this time, there's no miracle product for oral hygiene capable at long term to take over from the tooth brush and paste. The use at long term of chemicals in the bacteria plaque control can't be recommended because of their following effects and their slight effectivity-cost ratio. Nevertheless, oral hygiene chemicals combined with mechanical treatment provide very good results. Particularly, when used in the form of subgingival irrigation or slow liberation materials set in the periodontal pocket. PMID- 15536722 TI - Identifying the real issue. PMID- 15536723 TI - Solovy's insights. PMID- 15536724 TI - Designed to fail. PMID- 15536725 TI - The spoken vs. the written. PMID- 15536726 TI - The case for electronic records. PMID- 15536727 TI - Finance. Risky business. PMID- 15536728 TI - Patient care. Shortening the wait. PMID- 15536729 TI - Payment. Inside cuts. PMID- 15536730 TI - Data Page. Fix the spine and the bottom line. PMID- 15536731 TI - Legal. Fraud crackdown. PMID- 15536732 TI - Facilities. Take a left on oak. PMID- 15536733 TI - Legislation. Pushing for safety. PMID- 15536734 TI - Patient care. A helping hand. PMID- 15536735 TI - Hurting for healing. PMID- 15536736 TI - The open organization. Interview by Mary Grayson. AB - Richard Umbdenstock, chair-elect of the AHA, believes that the advent of benchmarking and comparative databases is one of the best things to happen to hospitals. And sharing that data and other information with the public will strengthen hospitals' relationships with their communities, says the CEO and president of Providence Services, Spokane, Wash. PMID- 15536737 TI - 2004 AHA environmental assessment. PMID- 15536738 TI - 5 years after IOM ... the evolving state of patient safety. AB - How much progress has the health care field made in its patient safety endeavors in the five years since "To Err Is Human" sparked major headlines and soul searching? PMID- 15536739 TI - From the storeroom to the boardroom. AB - As the nature of health care risk has changed, risk management has moved from an afterthought to major prominence in strategic, operational and financial planning. PMID- 15536740 TI - Dump the mumbo-jumbo. AB - Health literacy isn't just a problem for recent immigrants or people with little education. The arcane language providers use verbally and in writing confuses nearly everybody. PMID- 15536741 TI - The CEO and the future of technology. Panel discussion. AB - CEOs play a pivotal role in driving the selection and implementation of clinical and information technology within their organizations. Lack of standardization is viewed as the biggest obstacle to clinical transformation. PMID- 15536742 TI - Health insurance premiums climb five times faster than workers' pay and inflation. PMID- 15536743 TI - Chartbook maps women's health, shows state-by-state patterns for key indicators. PMID- 15536744 TI - Five chronic conditions account for one-third of health care spending growth. PMID- 15536745 TI - Gene discovery can lead to premature use of tests; shortage of genetic counselors likely, HealthTech warns. PMID- 15536746 TI - Injuries from medical devices sent more than 450,000 Americans to ED. PMID- 15536747 TI - Recasting IT strategy. Taking information technology out of the black box is a key to improving overall performance. PMID- 15536748 TI - Gradient elution in liquid column chromatography-prediction of retention and optimization of separation. PMID- 15536749 TI - Supercritical fluids for off-line sample preparation in food analysis prior to chromatography. PMID- 15536750 TI - Correspondence between chromatography, single-molecule dynamics, and equilibrium: a stochastic approach. AB - We report a detailed study concerning the correspondence between separations by chromatography, dynamic quantities coming from single-molecule measurements at the interfaces, and phase partition equilibrium by using the unifying approach of the stochastic description. The fundamental hypotheses allowing establishing the proper links between the three experimental techniques are discussed, and the full correspondence between the different quantities is determined from basic principles. The expressions of the errors on the retention factor which are intrinsically linked to the separation process, and which arise from peak splitting, peak tailing, stochastic bias, injection step, and number of the analyte molecules, are derived under general conditions and discussed in detail. Reference is made to the growing area of microsystems or nanosystems and chip technology, with numerical examples. How to determine the impact of single molecule dynamics observations on the chromatographic peak shape of the experimentally observed sorption time distribution and, in general, of the behavior of the species at the stationary phase (surfaces, interfaces) is pointed out. PMID- 15536751 TI - Solid-phase microextraction: a new tool in contemporary bioanalysis. PMID- 15536752 TI - Polyelectrolytes as stationary phases in liquid chromatography. PMID- 15536753 TI - Story teller or story analyst? How useful is the storied narrative for a critical sociology of CAM and nursing? AB - Despite recent developments in the sociology of complementary and alternative medicine (CAM), a critical analysis of the apparent affinity between CAM and nursing has, to date, remained essentially undeveloped. An empirical project is currently being conducted as an initial step to address the absence of such important critical research. A total of 30 written life history narratives were obtained from nurses working with and using CAM to explore such matters as professional boundaries and nurses' authentication strategies and conceptualisation and operationalisation of CAM. This paper addresses questions and conflicts that arose as the analytical tools were considered for these narrative accounts. Specifically, the paper explores whether the storied narrative sits easily with a critically oriented sociology of CAM; the differences between the role of "storyteller" or "story analyst"; and ask whether there is potential for developing a critical sociology of CAM nursing that retains the essence of personal stories. PMID- 15536754 TI - An empirically-based model for clinician-managers' behavioural routines. AB - Numerous past articles, many of which consist of idealised prescriptions for success or the occasional case study or practitioner's contribution, have commented on the role of hospital clinician-managers. Prior work is circumscribed, however, in that it tends to be normative and a priori (how clinician-managers in principle should manage) rather than descriptive and a posteriori (how clinician-managers in situ do manage). In addition, it is apparent that an empirically-grounded, testable model is lacking for the way clinician-managers work. This paper sets out to balance past normative prescriptive accounts with a descriptive-analytic one, and presents an empirically-based conceptual model of the behavioural routines of hospital clinician-managers. The model, based on multiple studies of clinician-managers' activities, conjectures five major modes of operating and four primary and five secondary pursuits. The paper advances accounts of how clinician-management work is conducted and the time frames for it, and hypothesises about clinician managers' relationships, and how power and control is experienced and exercised. It also briefly discusses some of the implications of both the research program and the findings. However, following Popper, researchers ought to invite attempts to improve rigor through a systematic critique of their findings. Critical analysis of this work under falsification processes is consequently welcomed PMID- 15536756 TI - Implementation of hospital restructuring and nursing staff perceptions of hospital functioning. AB - Hospital restructuring and downsizing has taken place in most developed countries during the past ten years. A small but growing body of research findings has identified aspects of these changes that serve as sources of stress for nursing staff, as well as features of the restructuring process that contribute to more effective transitions. This research reports results of a longitudinal study of hospital restructuring and downsizing on nursing staff perceptions of hospital effectiveness. Data were collected in November 1996 and again in November 1999 from nursing staff using questionnaires. Would the way hospital restructuring was implemented and managed in 1996 be related to nursing staff perceptions of hospital functioning in 1999? The findings showed positive but moderate relationships between more favorable restructuring processes and perceptions of hospital functioning. PMID- 15536755 TI - Medical managers' beliefs about reduced-hour physicians. AB - This paper examines medical managers' beliefs about the impact reduced-hour career paths for physicians has on organizational effectiveness. The findings of this exploratory inductive study of 17 medical managers at nine medical organizations in the Boston area suggest that managers believe the benefits of reduced-hour physicians (RHPs) far outweigh the disadvantages. However, many of their reasons appear to be exploitative of RHPs. In particular, managers believe that employing RHPs results in increased managerial control and that RHPs should: work more than they are compensated for; do a disproportionate share of the undesirable work; and remain extra flexible and available to the organization. An interpretation of the findings based on psychological contract theory is offered, and may help to illuminate other results reported in the literature, including some controversial findings that reduced-hour workers tend to have poor health outcomes. PMID- 15536757 TI - Accountability framework for managing stakeholders of health programs. AB - Understanding the incentives of stakeholders and employing effective management practices with various stakeholder groups is essential for program sustainability. This paper offers a conceptual model that depicts four different types of stakeholder interests that are relevant to health service organizations. The study identified the major stakeholders of telehealth programs, compared the influence of stakeholders by organizational ownership and investigated the practices used to manage these stakeholders. Quantitative and qualitative analyses demonstrated empirical support for the stakeholder model. The paper suggests that the model can be used as a stakeholder management tool in healthcare organizations and offers an assessment of the utility of the stakeholder framework in organizational research PMID- 15536758 TI - Perception of risk: a study of A&E nurses and NHS managers. AB - This exploratory study investigates perceptions of two groups of NHS staff, of a range of risks, topical and relevant to accident and emergency (A&E). Literature suggests that the effectiveness of a risk management strategy is lessened if staff exposed to risk, and managers, have different perceptions of the nature and severity of the risk. A self-administered questionnaire was distributed to registered A&E nurses and to Trust managers at directorate level and above, working in five large inner-city NHS Trusts. Managers and nurses were asked to quantify their perceptions of nine defined adverse events. They were also asked to identify the jobholder with day-to-day responsibility for managing four specific risks. Completed forms were obtained from 38 per cent of managers and nurses. Results showed broadly that managers tended to quantify risks of violence and stress to A&E nurses less highly than did nurses. By contrast, they tended to quantify risks to patients more highly than did nurses. Overall, a coherent pattern of difference in risk perception between the two groups was identified. It is argued that identification of differences of perception is an essential part of the risk management process. Illustrations are given of ways in which these differences can undermine the effectiveness of a risk management strategy. PMID- 15536759 TI - Talking to health managers about change: heroes, villains and simplification. AB - This paper presents contextual and empirical analyses of the pressures on senior health managers in the NHS through an examination of the documents produced by the NHS National Co-ordinating Centre for Service Delivery and Organisation on "Managing change". The result is a discussion and advocacy of the means through which "knowledge" with "knowing" might be brought together in the NHS to move it more towards an evidence-based approach to management. PMID- 15536760 TI - Evidence-based management in the NHS: is it possible? AB - The rise of evidence-based medicine and more recently evidence-based policy reflect the increasing importance of evidence as a basis for the organisation and delivery of health care. Evidence-based practice is central to the "modernisation" of health care in current UK policy. The latest manifestation of this process is the emergence of evidence-based management in health care. This paper examines the development of evidence-based approaches in health care and questions the appropriateness of such an approach to management. The problems inherent in applying the principles of EBP to management are explored and alternative apporoach based on the notion of craft is suggested as more practical and realistic. PMID- 15536761 TI - The impact of capitation on physician organizations' cost structures: an exploratory analysis. AB - This paper aims to evaluate the relationship between capitation payment methodology and the physician organization cost function. The paper provides evidence supporting a positive relationship between overhead rates and the level of capitation. Based on sample data, US medical practices whose net medical revenue consists of 11 percent or more capitation payments have significantly higher overhead costs per physician FTE. PMID- 15536762 TI - A performance improvement programme at a public hospital in Sri Lanka: an introduction. AB - Five-S, a simple tool was utilized as the initial step towards total quality management activities at a public hospital in Sri Lanka. This paper introduces the system improvement activities at the hospital which won several awards for quality of service at national level. Though there are multiple reasons for the significant improvement of performance at the hospital, the study team observes that Five-S has contributed heavily towards the success. The unique feature observed was that Five-S activity reorganizes the system radically compared to most of the continuous quality improvement (CQI) approaches which depend on problem solving. In the hospitals of developing countries, in which even the basic processes are unsatisfactory, Five-S approach may be suited for the initiation of the CQI process. Further research is needed to evaluate the quality improvement activity based on standardized criteria and to assess the factors which influenced the process. PMID- 15536763 TI - Organizational and management strategies in response to U.S. market trends. AB - This paper describes major trends in the health care market. They include increased health care costs, the growth of managed care, emphasis on quality of care, consumer choice and the growth of the elderly and uninsured populations. The relationship between cost, quality, managed care and choice are explored in the Medicare and Medicaid programs. A clearer understanding of these trends enables managers in health care organizations to make strategic decisions resulting in organizations' survival and growth. PMID- 15536764 TI - Acetaldehyde: deja vu du jour. AB - OBJECTIVE: The possibility that acetaldehyde is responsible for some of the central nervous system effects of ethanol has been a popular hypothesis for many years. This review examines the evidence of a role for acetaldehyde in the actions of ethanol in the brain. METHOD: The literature review was confined primarily to effects of acetaldehyde in the central nervous system in the realization that a great deal of information is also available on the actions of acetaldehyde in the periphery. The emphasis is on more recent findings, with only occasional references to older work. RESULTS: There are studies implicating acetaldehyde in nearly every central nervous system effect of ethanol that has been studied. With a few exceptions, the evidence for most of these effects is conflicting. For many years the dogma was that the brain did not metabolize ethanol. Any effects of acetaldehyde were therefore held to be due to acetaldehyde diffusing in from the blood. Recently, however, it has been established that ethanol is metabolized to acetaldehyde (primarily by catalase) and then to acetate (by aldehyde dehydrogenase) in the brain. These findings remove the problem that acetaldehyde does not penetrate the brain very well but leave questions as to what it does there. Almost invariably, the concentrations of acetaldehyde in the brain, under normal conditions of ethanol intoxication, are in the low micromolar range. Inhibition of aldehyde dehydrogenase will lead to increases of both peripheral and central acetaldehyde and usually to increases in the effects of ethanol or to behaviorally aversive effects. Stimulation of catalase should lead to increased levels of acetaldehyde in the brain, but this has not been directly demonstrated. Inhibition of catalase should lead to decreased acetaldehyde concentrations in vivo, but, again, this has not been directly demonstrated. Various effects of the direct application of acetaldehyde to the brain have been noted, but in most studies the concentration of acetaldehyde resulting from such manipulations has not been determined, and it is probably higher than that occurring during ethanol intoxication. These experiments tell us what acetaldehyde is capable of doing, not what it does after administration of ethanol. Still, this is a first step. CONCLUSIONS: Acetaldehyde is a product of ethanol metabolism in the brain. It clearly has central nervous system effects in its own right. The jury is still out as to whether it has effects under normal conditions of ethanol intoxication. This will remain the case until direct measurement of acetaldehyde concentrations in the brain is routinely accomplished under conditions in which behavioral effects of ethanol are also measured. PMID- 15536765 TI - Absinthe: attention performance and mood under the influence of thujone. AB - OBJECTIVE: The aim of this study was to determine whether the impacts of absinthe on attention performance and mood were different from those experienced with beverages that contain only alcohol. The ingredient causing absinthe's toxicity is believed to be thujone. METHOD: A total of 25 healthy subjects participated in the study. An attention performance test and two questionnaires testing different mood dimensions were used. Three drinks with an identical amount of alcohol but with different amounts of thujone were offered. RESULTS: The results of the present study showed that the simultaneous administration of alcohol containing a high concentration of thujone had a negative effect on attention performance. Under this condition, the subjects tended to direct their attention to signals in the central field of attention and to neglect peripheral signals; the number of correct reactions decreased significantly in the peripheral field of attention, and reaction time and the number of "false alarm" reactions increased significantly. The effects were most prominent at the time of the first measurement. When the subjects were under the influence of alcohol or were administered both alcohol and a low thujone concentration, these effects were not observed. The assessment of mood state dimensions showed that the anxiolytic effect of alcohol was temporarily counteracted by a high thujone concentration. CONCLUSIONS: As they are apparently opposed to the effect of alcohol, the reactions observed here can be explained by the antagonistic effect of thujone on the gamma-aminobutyric acid receptor. Similar alterations were observed for the other mood state dimensions examined. PMID- 15536766 TI - Personality traits and drinking to cope as predictors of hazardous drinking among medical students. AB - OBJECTIVE: The purpose of this study was to describe the prevalence and development of drinking to cope and hazardous drinking among medical students and to examine whether various personality traits and drinking to cope predict hazardous drinking. METHOD: In a 6-year prospective study of a nationwide sample of medical students (N = 421) assessments were made by questionnaire at the beginning (T1) and at the end (T2) of each participant's tenure at medical school. A cohort of 272 medical students (56% women) from all medical schools in Norway participated at both T1 and T2. The questionnaires encompassed measures of personality characteristics (Basic Characteristic Inventory) and alcohol-use (hazardous drinking and drinking to cope). RESULTS: The levels of drinking to cope and hazardous drinking were not significantly different between T1 and T2, rising from 9.2% to 11.8% and from 17.7% to 19.2%, respectively. Hazardous drinking at T1 (odds ratio [OR] = 7.0; 95% confidence interval [CI]: 3.2-15.4) and level of control (personality trait) at T1 (OR = 0.7; 95% CI: 0.6-0.9) predicted hazardous drinking at T2 among all the students. Hazardous drinking (OR = 3.5; 95% CI: 1.4-9.0), control (OR = 0.7; 95% CI: 0.6-0.9) and drinking to cope at T1 (OR = 5.0; 95% CI: 1.0-24.1) independently predicted hazardous drinking at T2 among the men, whereas the only predictor among the women was hazardous drinking at T1 (OR = 42.1; 95% CI: 8.1-218.2). CONCLUSIONS: Drinking to cope should be targeted for preventive measures against hazardous drinking, particularly among men. The effects of personality and drinking to cope differ by gender and should be studied further. PMID- 15536767 TI - Alcohol outlets and child physical abuse and neglect: applying routine activities theory to the study of child maltreatment. AB - OBJECTIVE: The purpose of this study is to examine whether or not alcohol access in neighborhood areas is differentially related to substantiated reports of child physical abuse and neglect. METHOD: This cross-sectional ecological study uses spatial regression procedures to examine the relationship between the number of bars, restaurants and off-premise outlets per population and rates of child physical abuse and neglect in 940 census tracts in California, while controlling for levels of social disorganization, population density and county of residence. RESULTS: The number of off-premise outlets per population was positively associated with rates of child physical abuse (b = 3.34, SE = 1.14), and the number of bars per population was positively related to rates of child neglect (b = 1.89, SE = 0.59). CONCLUSIONS: These results suggest that alcohol access is differentially related to type of child maltreatment, with higher densities of bars being related to higher rates of child neglect, and higher rates of off premise outlets related to higher rates of child physical abuse. The findings suggest there is a spatial dynamic of neighborhoods that can result in child maltreatment and underscore the importance of examining the alcohol environment when developing programs to prevent child maltreatment. PMID- 15536768 TI - Ethnic differences in the estimates of children exposed to alcohol problems and alcohol dependence in the United States. AB - OBJECTIVE: This study was undertaken to estimate the number of children nationwide exposed to an adult who has alcohol problems (AP) or alcohol dependence (AD) and to examine ethnic differences in the number of children exposed. METHOD: The study used survey data from the 1995 National Alcohol Survey, which constituted a nationally representative sample of household members 18 years of age and older in the 48 contiguous states, were randomly selected, with oversamples of blacks and Hispanics. A total of 4925 subjects were interviewed, with a response rate of 77%. The questionnaire asked about AP and AD in the previous 12 months. Rates from the survey and data from the U.S. 2000 Census were used to calculate the estimates. RESULTS: Overall, 11.6 million children (16%) were exposed to one or more AP and 2.1 million children (3%) were exposed to AD. A higher proportion, nearly 19%, of both black and Hispanic children as compared with 14.5% of white children were exposed to AP. Significantly higher proportions of black (5.2%) and Hispanic (4.9%) compared with white (2.0%) children were exposed to AD. CONCLUSIONS: A large number of children are being exposed to at least one adult suffering from alcohol problems or dependence. Black and Hispanic children are disproportionately affected by these problems. PMID- 15536769 TI - Drinking on campus: self-reports and breath tests. AB - OBJECTIVE: Concern about excessive alcohol consumption by college students has been raised by surveys indicating that more than 40% of students are "heavy" drinkers. This definition is based on students' reports of consuming five or more drinks (four or more for women) on an occasion sometime during the past 2 weeks. The present survey examines the degree to which this 2-week 5+/4+ drink criterion characterizes a student's pattern of alcohol use, and whether a 5+/4+ criterion for a drinking occasion is a valid indicator of high blood alcohol concentration (BAC). METHOD: Students (N = 856, 70% male) were interviewed as they returned home between 10 PM and 3 AM. Students reported their drinking of the past 2 weeks and of the night they were interviewed, then provided breath samples to determine their BAC. RESULTS: Among the students in the sample classified as "heavy" drinkers on the basis of self-reports, 49% had zero BAC on the night they were interviewed. Those who reported consuming 5+/4+ drinks the evening of the interview had a mean BAC <0.08%. The distribution of BACs in the entire sample showed 74.4% of students had a BAC of zero and 11.8% had a BAC <0.05%. Very high BACs (i.e., > or =0.15%) were rare (1.3%). CONCLUSIONS: Self-reports of consuming 5+/4+ drinks on at least one occasion during the previous 2 weeks did not reliably identify a pattern of heavy drinking. Moreover, reports of 5+/4+ drinks on an occasion were not necessarily associated with high BACs. PMID- 15536770 TI - Emergency room injury presentations as an indicator of alcohol-related problems in the community: a multilevel analysis of an international study. AB - OBJECTIVE: This study describes and examines the development of surrogate measures of acute alcohol-related injury for use in the evaluation of community based prevention initiatives. METHOD: An international collaborative study of alcohol and injury, the Emergency Room Collaborative Alcohol Analysis Project (ERCAAP), provided a subset of data on 8580 emergency room (ER) presentations from five countries and 28 ER facilities. RESULTS: Presentations most likely to be alcohol related were those made between 12:00 AM and 4:59 AM (56%), on Fridays, Saturdays or Sundays (26%); and those among injured persons who were male (28%), who were aged between 18 and 45 years (24%) or who were unmarried (24%). Multilevel logistic regression models confirmed the significance of the above variables as predictors of alcohol involvement prior to the injury event. The strongest predictor variable was presentation between 12 midnight and 4:59 AM with an odds ratio of 4.92 (Wald Test chi2 = 397.6, p < .001). Being male had an odds ratio of 3.01 (Wald Test chi2 = 247.25, p < .001), and presenting on a Friday, Saturday or Sunday night had an odds ratio of 1.50 (Wald Test chi2 = 49.6, p < .001), whereas being under 45 (odds ratio [OR] = 1.20, p < .05) and being unmarried (OR = 1.2, p < .01) were less strong predictors. Combining all these values for variables raised the probability of prior alcohol involvement in such injury presentations to 0.65, although only 3.37% of all cases met these criteria, limiting applicability of this combined variable as a surrogate measure for intervention studies. Probabilities of prior alcohol involvement are presented with other combinations of values for the predictor variables. CONCLUSIONS: Frequency of nighttime injury presentations to ER facilities, particularly by men, can be used as a reliable surrogate measure of alcohol related injuries for various epidemiological and evaluation purposes. PMID- 15536771 TI - Patterns of drug-related disorders in a prospective study of men chosen for their family history of alcoholism. AB - OBJECTIVE: Alcohol use disorders (AUDs) and abuse or dependence on illicit substances often co-exist. Most studies evaluating these phenomena have focused on less well-educated individuals, using cross-sectional designs. This study controls for several factors that might contribute to a propensity toward both alcohol and substance-related conditions in a prospective follow-up of middle class men. METHOD: The subjects (N = 249) were white sons of alcoholics and controls, followed for 20 years using structured interviews. They were evaluated at baseline and again 10, 15 and 20 years later. RESULTS: One in five subjects developed an illicit substance-related disorder by the 20-year follow-up, most commonly abuse or dependence on cannabinoids and/or stimulants. The men with these diagnoses were more likely to be recent smokers, to also fulfill criteria for alcoholism and to have a family history of a stimulant use disorder. They were not more likely to have family histories of alcoholism, did not have a lower response to alcohol at age 20 and had no enhanced risk for independent psychiatric disorders, although they did have relatives with manic depressive disease. CONCLUSIONS: Disorders related to illicit substances were common in this well-educated, highly functional population. Substance-related comorbidity with alcoholism did not seem to reflect several factors known to predict AUDs (e.g., family history of alcoholism and a low response to alcohol). The diagnostic overlap between AUDs and illicit substance-related disorders might reflect concomitant family histories of alcoholism and substance-related conditions and, perhaps, family histories of manic depressive disease. PMID- 15536772 TI - Alcohol-related advice for Veterans Affairs primary care patients: Who gets it? Who gives it? AB - OBJECTIVE: Most patients who misuse alcohol do not receive alcohol counseling from their providers. This study evaluated primary care patient and provider characteristics associated with receipt of alcohol-related advice and whether patients were advised to drink less or to abstain. METHOD: Outpatients from seven Veterans Affairs (VA) general medicine clinics were eligible if they screened positive for alcohol misuse, completed the Alcohol Use Disorders Identification Test (AUDIT) and answered questions about alcohol-related treatment and advice. Hierarchical logistic regression was used to evaluate patient and provider characteristics associated with patient reports of alcohol-related advice from a primary care provider in the past year. RESULTS: Among 5191 patients with alcohol misuse in the past year, 1554 (30%) reported receiving alcohol-related advice from their primary care provider during that time. Of patients advised, 73% reported advice to abstain. The likelihood of reporting advice increased as AUDIT scores increased: from 13% of patients with AUDIT scores <8 to 71% of those with scores > or =20. After adjustment for important confounders, measures reflecting the severity of alcohol misuse were most strongly associated with receipt of alcohol-related advice. Adjusted analyses also revealed increased odds of receiving advice among patients who reported liver disease, hypertension, current smoking or continuity of care. No measured provider characteristic was associated with giving advice in the fully adjusted model. CONCLUSIONS: This multisite VA study found that most patients with alcohol misuse did not receive alcohol counseling from a primary care provider. Moreover, providers predominantly offered advice to abstain, and they appeared to focus on patients with the most severe problems due to drinking or medical contraindications to drinking. PMID- 15536773 TI - The efficacy of a brief alcohol intervention combined with %CDT feedback in patients being treated for type 2 diabetes and/or hypertension. AB - OBJECTIVE: Alcohol biomarkers are being developed to improve a physician's ability to identify and intervene with patients with chronic medical problems adversely affected by heavy alcohol use. This article reports the findings of a brief intervention trial which included feedback to patients of their carbohydrate-deficient transferrin (CDT) test results. METHOD: A pilot study was conducted to test the efficacy of brief clinician advice to reduce alcohol use and improve health status in a sample of 151 patients being treated for Type 2 diabetes and hypertension. The intervention included informing patients of their CDT levels. The patients were randomized to a usual care or brief intervention group. RESULTS: There were no significant differences at baseline between the two groups in alcohol use, CDT levels, addiction rates, age, gender, socioeconomic status or health status measures. Following brief intervention, significant differences were observed in the intervention group in alcohol use and CDT: The proportion of heavy drinkers at the 12-month follow-up compared with baseline decreased from 35.8% to 24.7% in the intervention group, with no change in the control group (p < .044). CDT levels decreased as well from 2.79% to 2.41% (16% change) in the control group and 3.05% to 2.35% (28% change) in the intervention group, with significantly more intervention-group patients reducing their CDT level by at least 25% (p < .006). CONCLUSIONS: The study provides new information suggesting brief intervention, combined with feedback on CDT levels, can reduce alcohol use and %CDT in a sample of primary care patients being treated for Type 2 diabetes and hypertension. PMID- 15536774 TI - Gender differences in treatment outcomes for alcohol dependence among older adults. AB - OBJECTIVE: This study examined clinical characteristics and treatment outcomes of older alcohol-dependent men and women in a mixed-age private outpatient chemical dependency program. METHOD: The sample comprised 92 patients aged 55 to 77 (63 men and 29 women). The measures consisted of demographic characteristics, alcohol and drug use and dependence, drinking history, health status, psychiatric symptoms, length of stay in treatment, use of Alcoholics Anonymous and 6-month treatment outcomes. RESULTS: The women reported later initiation of heavy drinking (5+ drinks per occasion) than the men, but had similar drinking levels at the treatment intake interview. At the 6-month follow-up, 79.3% of women reported abstinence from alcohol and drugs in the prior 30 days versus 54.0% of men (p = .02). Greater length of stay in treatment predicted abstinence at 6 months. Among those who were not abstinent, none of the women reported heavy drinking in the 30 days prior to follow-up, whereas nonabstinent men reported a mean (SD) of 4.0 (9.2) heavy drinking days (p = .025). CONCLUSIONS: The results suggest that older women may have better drinking outcomes compared with older men, following treatment for alcohol dependence. PMID- 15536775 TI - Outcomes of substance use disorder treatment in suicidal and nonsuicidal male patients. AB - OBJECTIVE: Numerous studies report high rates of substance-related problems and psychopathology in substance use disorder (SUD) patients with a history of a suicide attempt. However, little is known about the response of suicidal SUD patients to treatment. This study examined the treatment outcomes of suicidal and nonsuicidal SUD patients who were followed for 5 years. METHOD: A total of 2099 male SUD patients were recruited from 15 Department of Veterans Affairs residential alcohol and drug treatment programs and were assessed at four points (treatment entry, discharge, and 1 and 5 years later). Approximately 7% (n = 156) of the patients reported a suicide attempt in the 3 months prior to the start of treatment. RESULTS: Although patients with a recent suicide attempt reported severe patterns of alcohol use and elevated psychiatric symptoms at baseline, they showed significant improvements in both of these domains at discharge from residential treatment, and these improvements were still evident at 1-year and 5 year follow-ups. Suicidal SUD patients were no more likely to leave treatment early than were nonsuicidal patients, and they received slightly longer and more individualized treatment. CONCLUSIONS: Despite a more severe pattern of alcohol use and psychiatric symptoms at baseline, suicidal SUD patients benefitted substantially from residential SUD treatment. These findings imply that suicidal SUD patients can be treated effectively within SUD treatment settings. PMID- 15536776 TI - Disulfiram therapy in patients with hepatitis C: a 12-month, controlled, follow up study. AB - OBJECTIVE: Although abstinence slows liver injury in alcoholic Hepatitis C (HCV) infected patients, few clinicians prescribe disulfiram because of concern over its hepatotoxic effect. Finding no controlled studies on this effect, we investigated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) patterns in seropositive (HCV[+]) and seronegative (HCV[-]) patients who received supervised disulfiram over 12 months. METHOD: We recorded retrospective aminotransferase measurements from medical records of 26 HCV(+) and 20 HCV(-) cases receiving 1500 mg disulfiram weekly in divided doses. Within groups, paired mean AST and ALT levels at 3, 6, 9 and 12 months were compared with baseline; between groups, nonpaired mean comparisons were used. RESULTS: There were no statistically or clinically significant elevations for the HCV(+) group at any time point. Between-group means were identical at all time points. CONCLUSIONS: Although sample size and retrospective design invite replication, the data suggest that disulfiram may be useful for HCV(+) alcohol-dependent patients in slowing hepatic injury by eliminating alcohol use and thereby removing the purported alcohol-HCV hepatotoxic synergy. It may also help to establish the abstinence criteria necessary to qualify for antiviral treatment. If disulfiram is used in HCV treatment, AST and ALT must be monitored closely. PMID- 15536777 TI - The role of behavioral undercontrol in the relation between alcohol use and partner aggression. AB - OBJECTIVE: Recent studies suggest that pre-existing aggressive personality traits moderate the relationship between alcohol use and intimate partner violence. The current study extends this literature by examining the relationship between behavioral undercontrol, patterns of alcohol use and relationship aggression in the context of a large, high-risk sample. METHOD: Participants were drawn from a cohort of 489 young adults taking part in an 11-year longitudinal study. Participants completed self-report measures of behavioral undercontrol, alcohol use and relationship aggression at Years 7 and 11 of the study when most were 25 and 29 years old, respectively. RESULTS: Results revealed that a composite measure of behavioral undercontrol predicted relationship aggression cross sectionally. Although the composite behavioral undercontrol measure did not interact with alcohol use to predict relationship aggression, one of the measures that comprised the composite (the Agreeableness scale from the Neuroticism Extraversion-Openness to Experience Five Factor Inventory) did. Longitudinal analyses revealed that behavioral undercontrol measured at Year 7 did not predict relationship aggression measured at Year 11. CONCLUSIONS: Results suggest that behavioral undercontrol is related to relationship aggression outside the laboratory. Individuals with low levels of agreeableness also appear to be at heightened risk for intoxicated aggression. Potential explanations for these findings are discussed. PMID- 15536778 TI - Exploring variability and sources of measurement error in alcohol expectancy questionnaire reliability coefficients: a meta-analytic reliability generalization study. AB - OBJECTIVE: Reliability Generalization, a relatively new meta-analytic score reliability technique, was used to examine reliability coefficients for Alcohol Expectancy Questionnaire (AEQ) subscales in published research. Specifically, the present study identified the typical reliability coefficients of scores on AEQ subscales across published studies and examined sources of measurement error across AEQ subscales and studies. METHOD: Based on literature reviews of PsyclNFO and PubMed, a total of 71 studies were analyzed and coded on 10 different dimensions believed to affect score reliability. RESULTS: Only 37% of the studies surveyed reported reliability coefficients for the data in hand. Analysis of these studies revealed that the average score reliability across studies varied considerably in subscales and samples, with the Arousal/Interpersonal Power and Sexual Enhancement subscales evidencing the greatest variability. Gender homogeneity and racial homogeneity were found to be the two most important predictors of the magnitude of score reliability coefficients across subscales and studies. CONCLUSIONS: Results of the study indicate that the AEQ tends to generate reliable scores, with some noted exceptions. Because the Arousal/Interpersonal Power and Sexual Enhancement subscales tend to generate unacceptably low reliability coefficients and because the AEQ holds promise in furthering prevention and treatment outcome research, it is suggested that future research with the AEQ explore instrument factor structure and item consistency. Because reliability estimates can influence substantive statistical analyses, it is recommended that authors using the AEQ report reliability information in their published work. PMID- 15536779 TI - Discriminating between substance-induced and independent depressive episodes in alcohol dependent patients. AB - OBJECTIVE: The purpose of this study is to examine the accuracy with which the depressive episodes of alcoholics undergoing treatment are classified as substance-induced depression (SID) or independent major depressive disorder (MDD). METHOD: Participants were 95 patients (64 men) undergoing partial hospital treatment for alcohol dependence. Affective disorders were assessed at baseline and 6 weeks, 3 months, 6 months and 12 months later. RESULTS: Over one quarter of the depressive episodes originally diagnosed as SID were reclassified as MDD during the 1-year follow-up, due to a continuation of the depressive episode despite a period of prolonged abstinence from alcohol and drugs. Both history of MDD and lower levels of alcohol dependence were associated with diagnostic change from SID to MDD. CONCLUSIONS: These findings support the diagnostic complexity of these two types of depressive disorders and point to clinical features that may differentiate them. PMID- 15536780 TI - Perspectives. Flu worries highlight more than one public-health quandary. PMID- 15536781 TI - CMS stresses tight timelines for MMA implementation. PMID- 15536782 TI - The microbiological case for adjunctive therapy for periodontitis. AB - That chronic periodontitis is an infectious disease is now firmly established, and the primary role of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola is generally accepted. Treatment by mechanical means such as scaling and root planing or surgery generally results in significant clinical improvement but may not arrest the progress or recurrence of disease. Several studies have shown that the probability of achieving lasting stability as measured by the arrest of progressive attachment loss and bone loss by primary mechanical therapy is a function, in major part, of whether pathogenic microorganisms are still present at local subgingival sites at the completion of active therapy. The infecting bacterial species are susceptible to killing by several antibiotics including, among others, tetracycline-class drugs, amoxicillin and metronidazole as well as by local exposure to chlorhexidine. Randomized clinical trials have shown that use of systemically administered antibiotics as an adjunct to mechanical therapies significantly enhances clinical outcomes and stability. Several slow-release devices that deliver anti-microbial drugs directly into periodontal pockets have been developed and are now on the market. Use of these devices permits local delivery of long-lasting, high concentrations of doxycycline (Atridox) minocycline (Arestin), and chlorhexidine (PerioChip) directly into periodontal pockets. Although these devices differ with regard to ease of use, concentration of drug delivered and length of time high drug concentrations can be maintained, randomized clinical trials have shown that their use as an adjunctive treatment to scaling and root planing results in a significantly greater reduction of periodontal pocket depth and an average increase in clinical periodontal attachment level of about 0.8 mm. Gain in clinical attachment is greater in deeper pockets than in shallower pockets. Locally delivered adjunctive anti-microbial therapy is an effective means to enhance therapeutic outcomes. PMID- 15536783 TI - A post-marketing study of 2805 patients treated for periodontal disease with Arestin. AB - Recent studies have demonstrated the effectiveness of locally delivered antibiotics adjunctive with scaling and root planing (SRP) in patients with chronic periodontitis. Specifically, several studies have demonstrated the efficacy of minocycline microspheres (Arestin). The objective of the current study was to evaluate the use of Arestin in a study using private practices all over the United States and adopting a predefined protocol. Eight hundred ninety five dentists and 2805 patients participated in the largest reported trial in chronic periodontal disease in private practice. The protocol outlined that patients were to have SRP at baseline with one application of Arestin in all pockets > or = 5 mm, a recall visit three months later that included a second application of Arestin, and a final assessment after 6 months. One thousand ninety five patients were treated in accordance with the protocol, and 1710 patients returned for a second assessment but only received one therapeutic intervention. Mean pocket depth reduction from baseline in the 1710 patients was 1.82 mm (p < 0.0001) and for the 1095 patients at 6 months it was 1.94 mm (p < 0.0001). Similar results were obtained in smokers, diabetes patients and patients with a history of cardiovascular disease. After 1 treatment 62% of sites had decreased to less than 5 mm and after two treatments the corresponding number was 67%. There were no serious adverse events in the study. This study demonstrated that a large study could be conducted in a private practice setting, that Arestin and SRP were effective in reducing pocket depth, and that adherence to the protocol yielded additional benefits. PMID- 15536784 TI - Treatment of incipient peri-implant infections using topical minocycline microspheres versus topical chlorhexidine gel as an adjunct to mechanical debridement. AB - This report presents the clinical results three months after application of minocycline microspheres as an adjunct to mechanical treatment of incipient peri implant infections compared to adjunctive treatment employing 1% chlorhexidine gel application. Sixteen patients in the minocycline group and 14 in the chlorhexidine group completed the study. Each patient had one or more implants with probing depth > or = 4 mm combined with bleeding and/or exudate on probing and presence of putative pathogenic bacteria. At baseline, patients were randomly assigned to minocycline or chlorhexidine treatment. Follow-up examinations were carried out after 10, 30, 60 and 90 days. The combined mechanical/antimicrobial treatment for the chlorhexidine group did not result in any reduction in probing depth and only limited reduction of bleeding scores. The adjunctive use of minocycline microspheres, on the other hand, resulted in improvements in both probing depths and bleeding scores. For the deepest sites of the treated implants, mean probing depth was reduced from 5.0 mm to 4.1 mm. The reductions in bleeding scores, although greater than for the chlorhexidine group, were modest. Thus, the question as to what extent the combined mechanical/minocycline treatment could be considered adequate for the treated lesions remains to be answered. The present short-term findings, however, encourage further studies with longer observation intervals on adjunctive use of minocycline microspheres in the treatment of periimplant lesions. PMID- 15536786 TI - Quality of care and performance improvement: an important new emphasis whose time has come. PMID- 15536785 TI - Periodontal treatment by Arestin and its effects on glycemic control in type 1 diabetes patients. AB - Studies indicate that a dual pathway between diabetes mellitus and periodontal disease exists. Elimination of periodontal infection by using systemic antibiotics in conjunction with scaling and root planing (SRP) improved metabolic control in diabetic patients, as defined by reduction in glycated haemoglobin or reduction in insulin requirements (Grossi and Genco, 1998). The aim of this randomised pilot clinical trial was to determine if type 1 diabetes patients with periodontitis will experience a reduction in HbA1c levels when treated with locally delivered minocycline microspheres (Arestin) as an adjunct to scaling and root planing. Twenty adult patients with poorly controlled diabetes (HbA1c 7.5%) and adult periodontitis, as determined by the presence of four teeth with 5 mm periodontal pockets, two of which had 6-9 mm pockets and bleeding on probing, were included in the study. All patients received full mouth SRP at baseline. Arestin was administered to all pockets > or => or = 5 mm at baseline and again at 12 weeks in the test group. Probing depth (PD), clinical attachment level (CAL), plaque index (PI), gingival index (GI), and HbA1c were evaluated at baseline and at weeks 6, 12, 18 and 24. The results demonstrated that local administration of Arestin as an adjunct to scaling and root planing is significantly more effective in reducing probing depths and providing a gain in clinical attachment levels than scaling and root planing alone in type 1 diabetic patients. Hb1Ac was reduced in all patients; however the difference between the test and control groups was not significant. PMID- 15536787 TI - Quality of care and health system performance: where are we now? Where should we be going? Who can take charge and lead us there? PMID- 15536788 TI - The practice of quality: incorporating high-yield strategies into the daily reality of medical practice. PMID- 15536789 TI - Data and information requirements for healthcare performance monitoring and improvement. PMID- 15536790 TI - Evidence-based medicine: ready for prime time? New and better. PMID- 15536791 TI - Evidence-based medicine: ready for prime time? Approach with caution. PMID- 15536792 TI - Disease management approaches to quality improvement. PMID- 15536793 TI - Educating a new generation of healthcare professionals with a lifelong commitment to quality of care. PMID- 15536794 TI - Making healthcare 'patient-centered': the centerpiece of quality improvement. PMID- 15536795 TI - Quality of care and performance improvement: two ideas that go hand-in-hand. PMID- 15536796 TI - Remembering Cecil. PMID- 15536797 TI - New directions in end-of-life and palliative care in North Carolina. PMID- 15536798 TI - Stem cell division. PMID- 15536799 TI - A job for Superman. PMID- 15536800 TI - Bad medicine. PMID- 15536801 TI - HSV-1 vs. HSV-2 demographics. PMID- 15536802 TI - Statistics puzzle. PMID- 15536803 TI - Diagnostic tests for HPV infection. AB - Human papillomavirus is one of the most common sexually transmitted infections. HPV-associated neoplasms, such as cervical cancer, are significant causes of morbidity and mortality worldwide. Recent advances in molecular biology have facilitated testing methods for HPV infection. Of the various methods, the only test cleared by the FDA for HPV testing of the cervix is the hybrid capture second-generation assay that uses RNA probes specific for the identification of certain high-risk or low-risk HPV types. Currently, the principal cervical-cancer screening modality is the Pap test, but given its higher sensitivity, HPV testing is being incorporated into algorithms for cervical-cancer screening. The development of HPV therapeutic and prophylactic immunization may expand the need for systematic HPV testing to help define eligible subgroups for intervention. PMID- 15536804 TI - The chemical pathology of insulin resistance and the metabolic syndrome. AB - In this review, we have examined the phenomenon of insulin resistance, a central manifestation of the metabolic syndrome. While it is by no means clear-cut, many new and exciting hypotheses have been proposed to explain this puzzling and enigmatic phenomenon. These studies have also led to a new way of looking at adipose tissue--it is no longer a passive repository of fat. It now actually appears to be a very active endocrine organ. A disturbance in this endocrine function helps contribute to the metabolic syndrome. PMID- 15536805 TI - Benchmark to a more successful lab operation. PMID- 15536806 TI - Create a no-blame lab environment. PMID- 15536807 TI - Misys' HIT unites care settings: CEO Tom Skelton tells how. PMID- 15536808 TI - Investigational test protocols must include informed consent. PMID- 15536809 TI - Petting zoo-associated Escherichia coli 0157:h7--secondary transmission, asymptomatic infection, and prolonged shedding in the classroom. PMID- 15536810 TI - No second chances. PMID- 15536811 TI - The midwife dismissed from Peterborough and Stamford Hospitals NHS Foundation Trust. PMID- 15536812 TI - The Discovery Health programmes, regarding birth and the care of the newborn. PMID- 15536813 TI - The Normal Birth issue. PMID- 15536814 TI - Blueprint for raising maternity standards NSF broadly welcomed, but midwives urge culture change. PMID- 15536815 TI - Midwifery basics: postnatal care: perineal repair. PMID- 15536816 TI - Let's hear it for the boys: male circumcision. PMID- 15536817 TI - Evaluating preterm breastfeeding training. PMID- 15536818 TI - Immunisation: a new approach. PMID- 15536819 TI - Developing a maternity unit visiting policy. AB - Southern Derbyshire Acute Hospitals NHS Trust has a policy of open visiting for people who wish to visit ward-based patients. The maternity unit, however, has adopted a slightly different approach, in which the visiting policy loosely consists of set hours with a degree of flexibility for mothers and visitors who wish to have extended visiting or want visitors outside the set hours. This article describes research undertaken to formulate a policy within the trust. PMID- 15536820 TI - Postnatal home visits: the parents' views. PMID- 15536821 TI - Hepatitis B vaccination: whose right? PMID- 15536822 TI - Independent at last. PMID- 15536823 TI - Pinning down history. PMID- 15536824 TI - What it takes to be a physician I.T. champion. PMID- 15536825 TI - Patient safety driving point-of-care I.T. plans. PMID- 15536826 TI - The metamorphosis of medical records. PMID- 15536827 TI - Mobile tech a hit with groups. PMID- 15536828 TI - CEOs talk shop: I.T. a strategic asset. PMID- 15536829 TI - Digital technology puts pen to computer. PMID- 15536830 TI - A little automation goes a long way. PMID- 15536831 TI - Sub-webs anything but sub-par. PMID- 15536832 TI - Readers perspective: "health care information technology consulting firms provide services that are worth the investment. Do you agree or disagree?". PMID- 15536833 TI - Microcracks in the alveolar bone following orthodontic tooth movement: a morphological and morphometric study. AB - Microcracks and microdamage have been associated with bone remodelling. The aim of this study was to investigate the role of microcracks as a trigger for alveolar bone remodelling after the application of an orthodontic load. In 25 3 month-old male Danish land-race pigs, the lower right first molar was moved buccally with a force of 130 cN. The contralateral molar was not treated and was used as an internal control. After 1, 2, 4, 7 and 15 days of treatment, the regions containing the right and left molars were excised and en bloc stained in basic fuchsin. The presence of microcracks on the buccal and lingual sides of both treated and untreated teeth was detected and expressed as crack density (number/mm2). The buccal treated side showed significantly more cracks than the buccal untreated side at day 1. This difference was significantly larger than that observed at days 2, 7 and 15. The same side showed significantly more microcracks than the lingual treated side at day 1, and this difference was larger compared with that observed at days 4 and 15. The presence of more microcracks on the treated side than on the untreated side suggests a role for microcracks in the initiation of bone remodelling after orthodontic loading. The increased presence of microcracks on the side towards which the tooth was moved, and where bone resorption is usually observed, suggests that microcracks could represent the first damage induced by orthodontic force that has to be repaired by bone remodelling. Finally, the strain levels in the alveolar bone during the orthodontic load transfer in the experiment were examined by finite element (FE) analysis. Although this showed that the strains were very low (10-100 microstrain), it should be noted that occlusal loading was not taken into account. In addition, high-resolution microtomography of the alveolar bone/periodontal ligament (PDL) interface revealed that the actual surface of the alveolar bone was very rough, predisposing it to high local stress/strain peaks. PMID- 15536834 TI - Effects of local administration of clodronate on orthodontic tooth movement and root resorption in rats. AB - Clodronate, a non-N-containing bisphosphonate, strongly inhibits bone resorption and also has anti-inflammatory properties. The purpose of this study was to examine the effect of the local administration of clodronate on orthodontic tooth movement. Both the right and left upper first molars of 26 7-week-old male Wistar rats were moved buccally with a standardized expansion spring. Clodronate solution was injected into the sub-periosteum area adjacent to the left upper molar every third day during the experimental period. The right first molar served as the control. Local injection of clodronate caused a significant (P < 0.001) and dose-dependent reduction in tooth movement in the rats. The number of osteoclasts on the clodronate-injected side was significantly less (P < 0.01) than on the control side. Local clodronate also inhibited root resorption incident to tooth movement. These results suggest that localized use of clodronate could be a useful therapeutic adjunct in orthodontic treatment. PMID- 15536835 TI - The efficacy of various alloplastic bone grafts on the healing of rat calvarial defects. AB - The objective of this study was to determine the relative efficacy of currently available alloplastic bone repair materials in the healing of rat calvarial defects histologically, histomorphometrically and biochemically. A representative material was selected from six major classes of bone repair materials and placed in 4 mm diameter calvarial defects of 6-week-old male Sprague-Dawley rats (five animals in the control and each of the six experimental groups). The outcomes were assessed after 2 months for alkaline phosphatase (ALP) activity and after 4 months of healing for histomorphometry. The tested alloplastic implant materials did not significantly increase ALP activity or the amount of new bone formation in the healing of rat calvarial defects relative to controls (P > 0.05). However, when the implant material itself was included in the analysis, significant differences were observed (P < 0.05). Additionally, the tested materials varied in their ability to bridge the bony defect. These data suggest that the rate of bone formation cannot be increased beyond control levels, rather the advantage of implant materials may be in their efficiency in filling the defect through incorporation of the material into the healing site and rapidly bridging the wound. PMID- 15536836 TI - The rat as a model for orthodontic tooth movement--a critical review and a proposed solution. AB - The aims of this study were to perform a systematic review of the use of rats as a model for experimental tooth movement, to give a critical evaluation of the use of elastics as a force delivery system, and to describe a newly designed well defined model for tooth movement in rats. The literature from 1981 to 2002 indicates that in 57 per cent of animal studies on orthodontic tooth movement, rats were used, but in many of these investigations the experimental set-up was poorly documented. Only three of the 159 studies fulfilled the inclusion criteria for a good model: a force magnitude of less than 20 cN; moving molar(s) mesially; an experimental duration longer than 2 weeks; and no extra experimental condition such as drug intervention. As more than one-quarter of the studies on tooth movement in rats used elastics to produce an orthodontic force, and as the forces they produced and their force decay during decompression are unknown, their mechanical characteristics on decompression were tested. Elastics stored under dry conditions or in water showed significant force decay from around 45 N to almost 0 N within the first 0.2 mm of decompression. With regard to the above mentioned shortcomings of using rats as a model for tooth movement, a newly designed experimental appliance for tooth movement in rats was evaluated. It proved to be stable and simple and able to deliver a continuous and constant force as low as 10 cN on all three molars together during an experimental period of 12 weeks without interference in animal welfare, and was able to compensate for the effects of molar distal drift and continuous incisor eruption. PMID- 15536837 TI - Root-crown ratios of permanent teeth in a healthy Finnish population assessed from panoramic radiographs. AB - An unfavourable root-crown (R/C) ratio caused by short dental roots may result from a developmental deficiency, root resorption after orthodontic treatment, or dental trauma. In the assessment of root shortening, subjective grading has often been used. For objective tooth measurements, varying materials and methods may make the results impossible to compare. This study used a simple, objective method to assess the R/C ratio (relative root length) of mature permanent teeth from panoramic radiographs (PRGs), tested its reproducibility and calculated the mean values of R/C ratios and their variations in a healthy Caucasian (Finnish) population. Two thousand seven hundred and seventy-nine teeth were measured on 108 PRGs. The intra- and inter-examiner reproducibility of the assessment method was good (Pearson correlation coefficients 0.87 and 0.83, respectively; P < 0.001) and the mean R/C ratios did not differ between the repeated measurements (P > 0.05). The biological variance in all cases exceeded the error variance for each tooth. These facts suggest that the method reported in this study can be used in the assessment of the relative root length of 'normal' teeth and its alterations in teeth with developmental or acquired aberrations of dental roots. Males, overall, tended to have higher R/C ratios than females; P-values varied from non-significant to less than 0.01. With the exception of the permanent lateral incisors in males and the permanent second molars in both genders, the ratios of the antagonist teeth were significantly greater in the mandible than in the maxilla (P < 0.05 for the lateral incisors of females; P < 0.001 for all other teeth). Consequently, in quantifying root shortening in developmentally short-rooted teeth, tooth- and gender-specific reference values should be employed. The Finnish R/C data reported here for all teeth except third molars could be used for comparison with other populations, patient groups or individuals where crown-root aberrations are suspected. PMID- 15536838 TI - Which factors influence willingness-to-pay for orthognathic treatment? AB - The aims of this interview-based questionnaire study were to establish which factors influence willingness-to-pay (WTP) for orthognathic treatment and to compare WTP values, from both members of the general public and orthognathic patients, with the actual cost of treatment, the hypothesis being that the more highly valued the intervention, the higher the WTP value. Data were collected from 88 orthognathic patients and a convenience sample of 100 adults using the so called 'payment card' method. Demographic data were recorded, as well as ability to pay, incisor relationship, occupation, and level of education. In addition, the resources used in orthognathic treatment were estimated for five patients who participated in the study. The results showed that there was a significant difference between the mean WTP values for the public and patient groups. Patients were prepared to pay [see symbol in tex]2750 more than members of the general public. In addition, a significant relationship was found between WTP and incisor relationship in the patient group, with Class II division 1 patients prepared to pay [see symbol in text]3130 more than those with Class III malocclusions. Ability to pay did not significantly affect WTP. The mean total costs estimated for orthognathic treatment were lower than the mean patient WTP value and similar to the mean WTP value for the public group. In terms of cost benefit, it appears that orthognathic treatment provides 'good value for money'. This study also showed that both patients and the general public were prepared to place a monetary value on the correction of dentofacial deformity and that this form of economic evaluation is a useful tool in monitoring health care in the UK. PMID- 15536839 TI - Dental aesthetics, self-awareness, and oral health-related quality of life in young adults. AB - The aim of the present study was to explore the putative relationship between dental aesthetics and oral health-related quality of life (OHRQoL), taking into consideration the potential direct and moderating influence of private and public self-consciousness. The subjects of this cross-sectional survey were 148 university students. Dental aesthetics were assessed by means of the aesthetic component (AC) of the Index of Orthodontic Treatment Need (IOTN). OHRQoL was estimated using a modification of the scales 'social appearance concern' and 'appearance disapproval', and a novel dental self-confidence scale. In addition, the private and public self-consciousness scales were used. Two-factor analyses of variance were carried out with high and low levels of dental aesthetics and private and public self-consciousness as the independent variables and the OHRQoL scales as the dependent variables. It was found that dental aesthetics had a direct effect on all OHRQoL scale values. Private self-consciousness was related to social appearance concern, while public self-consciousness was associated with both social appearance concern and appearance disapproval. An interaction effect was identified which showed that the impact of dental aesthetics on social appearance concern was stronger in respondents with high private and public self consciousness than in low scoring subjects. The findings of the study suggest that minor differences in dental aesthetics may have a significant effect on perceived OHRQoL. This effect was more significant in subjects with high self consciousness. PMID- 15536840 TI - Adaptive condylar growth and mandibular remodelling changes with bionator therapy -an implant study. AB - The purpose of this study was to describe condylar growth and mandibular remodelling changes associated with bionator therapy. Twenty-five patients (15 males and 10 females) between 6.9 and 11.2 years of age with Class II division 1 malocclusions were randomly allocated to either control (n = 11) or treatment (n = 14; bionator only) groups and followed longitudinally for approximately 1 year. Treatment consisted of a bionator only, constructed to clear the buccal dentition by 2 mm and to position the mandible into an edge-to-edge incisor relationship. Using metallic implants for superimposition, mandibular growth, displacement, and true rotation were evaluated cephalometrically. The results showed significant changes in the direction (more posterior) but not in the overall amount of condylar growth. The bionator appliance produced greater than expected posterior drift of landmarks in the condylar and gonial regions. Cranial base superimposition showed greater than expected anterior mandibular displacement, but little or no true mandibular forward rotation with bionator therapy. The bionator appliance alone produced changes in condylar growth direction and remodelling changes associated with mandibular rotation and displacement. PMID- 15536841 TI - A modified monobloc for the treatment of obstructive sleep apnoea in paediatric patients. AB - The aims of this study were to determine the differences in craniofacial morphology between children with obstructive sleep apnoea (OSA) and control subjects, and to investigate the effects of modified monobloc (MM) appliance treatment in OSA children. The treatment group consisted of 20 OSA Caucasian subjects (10 boys and 10 girls) with ages ranging from 4 to 8 years (mean 5.91 years). The control group comprised 20 healthy Caucasian subjects without OSA (10 boys and 10 girls) with ages ranging from 5 to 7 years (mean 6 years). Polysomnography was used to establish the diagnosis of OSA and to evaluate the effects of MM treatment in the test group. Cephalometric radiographs and study models were obtained for all subjects. A number of statistically significant differences were detected in craniofacial morphology between the treatment group and controls. The treatment group demonstrated a skeletal Class II pattern (P= 0.04), with a reduced mandibular length (P= 0.03) and a corresponding increase in overbite. The hyoid bone was located superiorly in the OSA group (P = 0.04). Analysis of the dental arches demonstrated a significantly smaller distance between the first and second inter-molar primary mandibular regions (P = 0.03 and P = 0.04, respectively) in OSA patients. Repeat polysomnography, with the MM in situ, after 6 months of wear, revealed a significant reduction in the apnoea hypopnoea index in children with OSA (P= 0.0003). The MM was found to reduce daytime sleepiness and to subjectively improve assessed sleep quality. The patients and their parents reported good compliance with MM treatment. PMID- 15536842 TI - Comparison of initial shear bond strengths of plastic and metal brackets. AB - The aims of this study were to compare initial and 24 hour shear bond strengths of plastic brackets with a mechanical base (Spirit MB) and metal brackets, using two different adhesives (System 1+ and Enlight), and to examine the modes of failure after debonding. Eighty extracted human premolars were used. After bonding, shear bond strengths in half the sample were tested within 30 minutes. The remaining 50 per cent were placed in a 37 degrees C distilled water bath for 24 hours before testing. The results showed that the effects of the two adhesives and the interaction of the two time intervals and the two bracket types on shear bond strength were significantly different (P < 0.05 and P < 0.0001, respectively). Six of the 10 groups were found to have less than 50 per cent of the adhesive remaining on the tooth surface after debonding in the 24 hour metal bracket-System 1+ group, but most specimens in the other seven groups had more than 50 per cent of the adhesive left. It is concluded that System 1+ cannot provide sufficient initial bond strengthfor Spirit MB and may increase the risk of enamel fracture for metal brackets. PMID- 15536843 TI - A comparative in vitro study of the strength of directly bonded brackets using different curing techniques. AB - The aim of this study was to compare, by shear testing, the bond strengths after 1 and 24 hours of a light-cured resin (Enlight) and a light-cured glass ionomer cement GIC (Fuji Ortho LC) using various polymerization lamps (halogen, high performance halogen, xenon, and diode) for the direct bonding of brackets. The self-curing resin (Concise) was used as the control. The analysis was carried out using the SPSS program. For group comparison purposes, the single factor variance analysis (ANOVA) and the post-hoc test (Tukey's HSD) were used. The level of significance was established at P < 0.05. When comparing two mean values the t test for independent random samples was employed. All polymerization lamps achieved the minimum bond strength of 5-8 MPa. With Enlight LV, bond strength was dependent on curing time (the halogen lamp achieved the highest bond strength of 10.0 MPa, P < 0.001, with a curing time of 40 seconds. The other lamps showed similar results) and on the mode of cure (the highest bond strength values were achieved by four-sided curing, P= 0.04). Fuji Ortho LC, on the other hand, was independent of the duration of light curing and the type of lamp used. The bond strengths of the resin-modified glass ionomer cement (RMGIC) were similar to or somewhat higher than those achieved with light-cured composite resin (P = 0.039) when lamps with short polymerization times were used, but were significantly lower (P< 0.001) when compared with the self-curing composite adhesive. After 24 hours, the bond strengths of all adhesives showed a significant increase: Enlight 19 per cent, Fuji Ortho LC 6.6 per cent, Concise 16 per cent. Bond failure occurred for Enlight at the bracket-composite resin adhesive interface in 90 per cent and with Concise in 57 per cent. However, Fuji Ortho LC showed far more cohesive and mixed failures, indicating an improved bond between bracket and cement. PMID- 15536844 TI - A comparative study of polymerization lamps to determine the degree of cure of composites using infrared spectroscopy. AB - The aim of this study was to compare polymerization lamps of various technologies (halogen, high performance halogen, xenon, i.e. plasma arc, as well as diode lamps) to determine the degree of polymerization of composite probes using Fourier-transform infrared (FTIR) spectroscopy. To observe the light-induced curing reaction under a bracket, an attenuated total reflection (ATR)-FTIR was used. The results showed that the recommended curing times for all the polymerization lamps tested were adequate for successful polymerization of the composite used (degree of conversion approximately 60 per cent, which did not exceed 66 per cent after longer curing times). However, even after very long curing times, delayed curing of the composite must be taken into consideration as there was a decrease of approximately 10 per cent in FTIR band intensities (1637 and 3104 cm(-1)). ATR-FTIR spectroscopy showed that when bonding brackets with materials cured by light, uncured areas remain in the centre of the lowest layer of the adhesive. PMID- 15536845 TI - The effects of argon laser curing of a resin adhesive on bracket retention and enamel decalcification: a prospective clinical trial. AB - A prospective clinical trial was carried out to compare argon laser-curing of a traditional light-activated composite resin with conventional visible light curing in terms of bond failure rate and incidence of enamel decalcification. Forty-five patients with a total of 742 metal brackets bonded to the upper and/or lower teeth in a modified split-mouth design participated in the study. The adhesive (Transbond XT) on the control teeth was cured by conventional visible light for 40 seconds, and the experimental teeth were exposed to 10 seconds of 250 mW argon laser irradiation. The patients were monitored for a period of 14 months. Intraoral photographic slides of the maxillary anterior teeth (212 in total) were taken at the beginning of treatment, after 12 months of treatment, following application of a plaque disclosing agent, and at the end of the observation period, following temporary debonding. Seven dentists used standardized rating systems to evaluate decalcification and plaque accumulation. The results of this study indicated that there were no significant differences between curing methods for the incidence of decalcification and plaque accumulation. However, the 10 second argon laser-curing method showed a statistically lower bond failure rate (2.4 per cent, P < 0.05) than the 40 second conventional visible light-curing method (5.7 per cent). It is concluded that the use of argon laser curing is superior to that of conventional light-curing with respect to bond failure and chairside time. However, the incidence of decalcification seems to be similar. PMID- 15536846 TI - Effect of sodium hypochlorite treatment on the molecular composition and morphology of human coronal dentin. AB - PURPOSE: The aim of the present study was to evaluate the deproteination potential of 5% aqueous NaOCl solution applied by rubbing action on the molecular composition and morphology of smear-layer covered and acid-etched human coronal dentin surfaces. MATERIALS AND METHODS: Paired specimens (n = 4 x 2 per group) of acid-etched (Group A) and smear-layer covered (Group B) human coronal dentin surfaces were sequentially treated with the NaOCl solution for time intervals ranging from 5 s to 120 s and analyzed by reflectance FTIR microspectroscopy and tapping mode atomic force microscopy. The changes in the mineral (v4 P-O stretching vibrations) to matrix (C = O stretching vibrations of amide I) peak area ratios and in the Ra roughness parameter were used to quantify the effect of treatment. Two-way ANOVA and Dunn's tests were used to assess the differences within each group and between groups (a = 0.05). RESULTS: In both groups, NaOCl treatment reduced organic matrix (amide I, II, III peaks), but did not affect carbonates and phosphates. In group A, the rate of deproteination was slow, and reached a peak value after 120 s. Tubule orifices became visible after 40 s of treatment; after 120 s, excessive porosity was detected, with Ra values presenting no statistically significant difference from group B. In group B after 10 s, the extent of deproteination was enhanced, reaching a plateau between 30 s and 60 s, and attaining a maximum after 120 s. Tubule diameter, intertubular porosity, and Ra were increased; intertubular dentin area was reduced. For both groups after 40 s of treatment, the mineral to matrix ratio recorded was similar to smear-layer-free sectioned dentin. CONCLUSION: The results of the present study imply that deproteination of mineralized or acid-etched dentin surfaces within a clinically relevant time frame may provide methods for bonding to dentin alternative to conventional technique-sensitive dentin hybridization. PMID- 15536847 TI - Ultrastructure of the dentin-adhesive interface after acid-base challenge. AB - PURPOSE: To observe the ultrastructure of the dentin-adhesive interface after in vitro sequential challenge by acidic and basic chemicals around adhesive restorations. MATERIALS AND METHODS: Box-shaped cavities were prepared in bovine root dentin and restored as follows: Reactmer Bond and Reactmer Paste (RB/RP), Clearfil SE Bond and Clearfil AP-X (SE/APX), ABF (experimental) and APX (ABF/APX), Single Bond and APX (SB/APX). After the specimens were stored in water for 1 week, the integrity of the bonds tested by sequential immersion were placed in an artificial demineralizing solution (pH 4.5) for 20 min and in 5% NaOCl for 20 min. The specimens were sectioned, polished, then argon-ion etched for 7 min, and gold sputter coated for SEM examination of the dentin-adhesive interface. RESULTS: The morphological results indicated tight bonding between the cavity wall dentin and the adhesive. For the fluoride-releasing restoration, RB/RP, a thick acid-resistant zone was clearly observed adjacent to the restoration; however, the bonding resin, RB, was partially degraded by the acid-base challenge. For SE/APX, ABF/APX and SB/APX, the bonding resins were resistant to the acid-base challenge. With the self-etching adhesive systems, SE and ABF, a thin band of acid-base resistant dentin less than 1 microm thick was observed beneath the hybrid layer. With the wet bonding system, SB, the hybrid layer was partially degraded by the acid-base challenge. CONCLUSION: SEM observation is useful for observing the reactions of adhesives to acid-base challenge. Prevention of secondary caries around a restoration may be influenced by the physical properties of the restorative and adhesive materials, quality of the hybrid layer, and fluoride-release. PMID- 15536848 TI - Influence of load cycling and tubule orientation on ultimate tensile strength of dentin. AB - PURPOSE: The purpose of this study was to evaluate the effect of mechanical load cycling and tubule orientation on the ultimate tensile strength of crown and root dentin. MATERIALS AND METHODS: Twenty bovine teeth were cleaned and their incisal surfaces reduced, resulting in a flat surface 4 mm above the cementoenamel junction. The teeth were divided into 4 groups: G1 = control (no mechanical load cycling); G2 = 1,000,000 load cycles at 50 N; G3 = 1,000,000 load cycles at 100 N; G4 = 1,000,000 load cycles at 200 N. Sections of 0.5+/-0.1 mm thick were cut mesiodistally and the slabs further trimmed to 0.5+/-0.1 mm in an hourglass shape, according to the tubule orientation (parallel and/or perpendicular to the long axis of the tooth) in the root and crown, with a fine diamond bur to produce a cross-sectional surface area of 0.25 mm2. All specimens were then subjected to ultimate tensile strength testing at a crosshead speed of 1 mm/min. Means and standard deviations were expressed in MPa. The bond strength data were analyzed with three-way ANOVA and Fisher's PLSD test (p < 0.05). RESULTS: Ultimate tensile strength values did not differ statistically significantly between the loaded group and the control. However, statistical differences existed between root and coronal dentin (p < 0.001) and between parallel and perpendicular orientation of tubules (p < 0.001), where root and sections parallel to the tubules presented higher values than coronal and perpendicular, respectively. These differences were not related to mechanical loading. CONCLUSION: There was no effect of in vitro load cycling on ultimate tensile strength of dentin. Differences presented between ultimate tensile strength of crown vs root dentin and parallel vs perpendicular to the tubule orientation can be influenced by differences in the amount of organic and inorganic content as well as composition of the dentin matrix. It is important to consider these aspects when predicting the quality and durability of restorations according to the types of dentin and tubule orientation to be restored. PMID- 15536849 TI - Effect of bur type on microtensile bond strengths of self-etching systems to human dentin. AB - PURPOSE: To compare the microtensile bond strength (microTBS) of five adhesives to human dentin prepared with 600-grit SiC abrasive paper (SiC), a diamond rotary instrument, or a carbide bur. The null hypothesis was that different cavity preparation instruments do not affect adhesion of resin adhesives. MATERIALS AND METHODS: Human molars (n = 45) were randomly divided into three groups according to surface treatment. Each group was bonded using a total-etch adhesive (Single Bond, 3M ESPE), one of three self-etching primer systems (Clearfil SE Bond or ABF, Kuraray; Imperva Fluorobond, Shofu), or a self-etching adhesive (One-Up Bond F, Tokuyama). A 4-mm composite crown was built over the bonded surface. Specimens were stored in water for 24 h at 37 degrees C. They were sectioned into 0.7-mm thick slabs, trimmed to a cross-sectional area of 1 mm2, and loaded to failure at a crosshead speed of 1 mm/min using a tabletop tester (EZ-Test, Shimadzu). Microtensile bond strength data were analyzed using analysis of variance and Fisher's PLSD test. RESULTS: Surface preparation using a carbide bur generally yielded higher bond strengths than preparation using either a diamond rotary instrument or SiC abrasive paper. SE Bond had the highest mean microTBS of the five adhesives tested. CONCLUSION: Resin-dentin bond strengths can be affected by the type of instrument used to prepare the tooth. Specifically, higher bond strengths might be achieved by using carbide burs rather than diamond cutting instruments. PMID- 15536850 TI - The effect of nonvital bleaching on the shear bond strength of composite resin using three adhesive systems. AB - PURPOSE: To evaluate the effect of nonvital bleaching on the shear bond strength of composite resin using three adhesive systems. MATERIALS AND METHODS: Two hundred seventy bovine teeth were assigned to 3 groups (n = 90): SP, paste of sodium perborate and water; CP, 37% carbamide peroxide; CO, control group (no treatment). After the bleaching treatment, the teeth in each group were cut into enamel (E) and dentin (D) sections. The teeth were embedded in polyester resin and polished in order to obtain a flat E or D surface. Each group was divided into 6 subgroups (n = 15) according to substrate (E and D) and adhesive system: SB, Single Bond (solvent: water/alcohol); PB, Prime & Bond NT (solvent: acetone); CLF, Clearfil SE Bond (solvent: water). The adhesive system was applied on each flat surface according to the manufacturer's instructions and a cylinder of composite resin Z-250 was overlaid. The specimens were stored in distilled water for 7 days at 37 degrees C. The SBS test was performed in a universal testing machine with crosshead speed of 0.5 mm/min. The data obtained (in MPa) were statistically analyzed with two-way ANOVA and Tukey's test (p < or = 0.05). RESULTS: The bleaching treatment influenced the SBS, regardless of the adhesive system used. On enamel, the SP and CP bleaching treatments reduced the SBS values. On dentin, the SP bleaching agent reduced the SBS values, but the CP bleaching did not. CONCLUSION: Nonvital bleaching treatment with sodium perborate may adversely affect shear bond strength of composite resin for both enamel and dentin. Similar effects can be expected from 37% carbamide peroxide bleaching agent on enamel but not dentin bonding strength. The use of water/alcohol and acetone-based adhesive systems cannot reverse the effects of bleaching treatments on bond strengths. PMID- 15536851 TI - In vitro sealing performance of two one-step adhesive systems in cervical cavities. AB - PURPOSE: To evaluate the effects of 1) wall grinding status, 2) thermal stress, and 3) time of polishing on sealing performance of two one-step bonding systems in cervical cavities. MATERIALS AND METHODS: Semicircular cavities with dimensions of 3 x 3 x 1.5 mm were prepared along the CEJ of extracted human premolars. In experiment 1, the cavity walls were additionally ground with round steel diamond burs (regularly ground) or finished with superfine diamond burs. The cavities were treated with AQ Bond (AQ) or One-up Bond F (OB) and filled with a resin composite. Then thermocycling (5 degrees C to 55 degrees C, 1000 x) was performed. In experiment 2, the regularly ground cavities were used. After restoration, the teeth were subjected to thermocycling (1,000 to 5,000 x). In experiment 3, the restorations in the regularly ground cavities were polished immediately or 24 h after filling. No thermocycling was carried out in this group. All specimens were immersed in a dye solution for 2 h. Microleakage at coronal and apical walls was evaluated on the longitudinal sections of the restorations. RESULTS: For OB, the cavities finished with superfine burs exhibited more microleakage at the coronal wall than the regularly ground cavity (p < 0.05). The leakage at the coronal walls increased with the number of thermal cycles for both systems (p < 0.05). The delayed polishing generated better adaptation at the apical wall of AQ and at the coronal wall of OB (p < 0.05). CONCLUSION: Grinding status, thermocycling, and time of polishing had influences on the sealing performance of the two one-step adhesive systems. PMID- 15536852 TI - Effect of thermal and mechanical load cycling on nanoleakage of Class II restorations. AB - PURPOSE: To evaluate the effect of thermal and mechanical cycling on the degree and pattern of nanoleakage on cervical margins of Class II restorations. MATERIALS AND METHODS: Forty box-type Class II cavities were prepared on bovine incisors. The cavities were restored with Single Bond and Z-250 composite resin (3M-ESPE) according to manufacturer's instructions. The teeth were randomly assigned to 4 groups: G1, control; G2, thermal cycling (2000 cycles, 5 to 55 degrees C); G3, mechanical load cycling (100,000 cycles, 50 N); G4, thermal and mechanical load cycling group (2000 cycles 5 to 5 degrees C/100,000 cycles, 50 N). The specimens were then sealed leaving a 1 mm window around the cervical margin interface. Samples were immersed in a 50% w/v ammoniacal silver nitrate solution for 24 h, and exposed to a photodeveloping solution for 8 h. Specimens were sectioned longitudinally, embedded in epoxy resin, polished and mounted on stubs, gold sputter coated, and examined under SEM using backscattered electron mode. Silver particle penetration length was measured directly on the SEM monitor and calculated as the percentage of the total length of cut dentin surface that was penetrated by silver nitrate. The data were analyzed with ANOVA and Fisher's PLSD test (p < 0.05). RESULTS: The degree of nanoleakage significantly increased when thermal and mechanical cycling was performed on the same specimens, as compared to the other groups (p < 0.05). No differences were observed between the control, thermal cycling, and mechanical cycling groups. No difference in nanoleakage pattern was observed between the groups. CONCLUSION: Thermal and mechanical cycling combined adversely affected nanoleakage values. Simulation of the oral condition might be crucial to better evaluate and understand the performance of adhesive materials. PMID- 15536853 TI - Influence of adhesive system and placement technique on microleakage of resin based composite restorations. AB - PURPOSE: To analyze the influence of two placement techniques and two adhesive systems on the marginal microleakage of Class II resin-based composite restorations using different resin composite materials. MATERIALS AND METHODS: Two hundred standardized Class II cavities (5 mm x 3 mm x 1.5 mm) were cut in human premolars and restored using five resin composite materials (A.L.E.R.T.-AL, Solitaire-SO, SureFil-SU, Filtek A110-A, and Z100-Z), two adhesive systems (Bond 1-B1 and Etch & Prime 3.0-EP), and two placement techniques (incremental or bulk). Specimens were thermocycled (700 cycles, 5 degrees - 55 degrees C, 1 min dwell time), immersed in 0.5% methylene blue solution for 4 h, and sectioned longitudinally to evaluate the extent of microleakage. Data were submitted to statistical analysis using the Kruskal-Wallis test. RESULTS: It was observed that both adhesive systems presented similar results (p > 0.05), and neither achieved complete marginal sealing. The bulk placement technique led to greater microleakage than the incremental technique (p < 0.001). Restorations prepared using SO resin composite showed greater microleakage than those performed using Z or AL materials. However, SO obtained similar values of microleakage when compared to SU and A materials. CONCLUSION: The use of incremental placement technique resulted in less microleakage for Class II resin-based composite restorations. No difference was observed among adhesive systems regarding prevention of marginal dye penetration. Furthermore, the type of resin composite material employed for restoration may have an influence on the extent of microleakage. PMID- 15536855 TI - Stabilization effects of CAD/CAM ceramic restorations in extended MOD cavities. AB - PURPOSE: Using extended, standardized MOD preparations, it was the aim of this in vitro study to examine the performance of CAD/CAM ceramic inlays in comparison to composite inlays after mechanical and thermal fatigue loading in terms of marginal quality and stabilization of the remaining tooth structure. MATERIALS AND METHODS: Standardized cavities with different wall thicknesses were prepared in 90 extracted premolars; 10 additional premolars remained untreated. Composite inlays (Tetric) and CAD/CAM restorations (Cerec II; Vita Mark II) were adhesively placed in the cavities. After loading in a chewing simulator, quantitative and qualitative marginal gap examinations were conducted and fracture resistance determined. RESULTS: The results show that ceramic inlays provide significantly greater stabilization and better marginal quality than do composite inlays. CONCLUSION: Chairside-fabricated ceramic inlays inserted using adhesive technology are able to stabilize weakened cusps. In the case of very thin remaining walls (about 1.3 mm), however, the marginal quality and the cusp stabilizing effect are also reduced. PMID- 15536854 TI - Effects of light intensity through resin inlays on the bond strength of dual cured resin cement. AB - PURPOSE: To evaluate the influence of the light intensity, irradiation time, and thickness of the indirect restoration composite on the bond strength of dual cured resin cement immediately after cementation, applying the resin coating technique. MATERIALS AND METHODS: Three hundred forty composite blocks as an adhesive surface and four thicknesses of indirect restoration composite disks were prepared. The surface of the composite blocks was coated with low-viscosity resin composite and immersed in water for 24 h. After immersion, dual-cured resin cement was applied to the resin-coated surface and the indirect restoration composite disks were placed on it. Light irradiation was performed through four thicknesses of indirect restoration composite disks with conventional halogen (40 s) and high intensity (10, 20, 40 s) light units. The specimens were stored in water at 37 degrees C for 10 min or 24 h, and the tensile bond test was performed. RESULTS: For the 1-mm composite thickness, there were no significant differences in the bond strengths between 10 min and 24 h after cementation with the four light curing methods. For the composite thicknesses of 3 or 4 mm, the bond strength 24 h after cementation was significantly higher than that at 10 min after cementation with conventional halogen (40 s) and high-intensity light units at 10 s and 20 s, but there was no significant difference with the high-intensity light unit applied for 40 s. CONCLUSION: The bond strength of dual-cured resin cement immediately after cementation could be greatly influenced by the irradiation time and the light intensity penetrating the indirect restoration composite. PMID- 15536856 TI - Clinically controlled study on the quality of class III, IV and V composite restorations after two years. AB - PURPOSE: The quality of fillings consisting of a hybrid composite was compared with fillings consisting of two different microfilled composites in a clinical examination. MATERIAL AND METHODS: Within the design of a clinically controlled two-year study conducted under practice-relevant conditions, the composites C Fill MH (Megadenta), Helio Progress (Vivadent), and Visio-Dispers (ESPE) were examined in anterior-tooth cavities in 134 patients by means of the split-mouth technique. The clinical examinations were performed based on modified Ryge criteria (marginal integrity, anatomical form, secondary caries, color, marginal discoloration, surface roughness) after 12 and 24 months. RESULTS: Immediately following filling application, all evaluation criteria except color were assessed as level A. After 12 and 24 months, C-Fill MH was found to be significantly superior to the other materials with regard to marginal integrity, color, and marginal discoloration. CONCLUSION: The hybrid composite C-Fill MH seems to be superior to the microfilled composites and should therefore be preferred as a restorative filling material. PMID- 15536857 TI - Clinical/photographic evaluation of a single application of two sealants after eleven years. AB - The clinical behavior of Concise and Prisma Shield sealants for pit and fissure was analyzed by clinical/photographic evaluation. A hematoxylin-based staining solution was applied to the occlusal surface 7 days, 18 months, 36 months, and 11 years after the occlusal sealing, thus allowing the sealant material on the surface to be checked. At each analysis time, each occlusal surface was photographed, and the photographs corresponding to each time were submitted to clinical/photographic evaluation. Although the sealant materials showed marginal alterations over time, they did not require reapplication because the region composed of the pit and fissure remained covered. PMID- 15536858 TI - Colonization by Porphyromonas gingivalis and Prevotella intermedia from teeth to osseointegrated implant regions. AB - Colonization by periodontopathic bacteria is a risk factor for peri-implantitis. The purpose of this study was to investigate the colonization by black-pigmented anaerobic bacteria that occurs between the time before fixture installation and 6 months after inserting superstructures in implant treatment in partial edentulous cases. Dental plaque was serially collected from around the natural teeth and implants in 12 patients in whom a dental implant was indicated, and Porphyromonas gingivalis and Prevotella intermedia were detected using polymerase chain reaction (PCR). One month after connecting the abutment, the detection rate of P. gingivalis per site from around the implants was 63.7% and that of P. intermedia was 50.8%. Six months after superstructure setting, the detection rate per site of P. gingivalis from around the implants was 56.8% and that of P. intermedia was 41.1%. When chromosomal DNA segmentation patterns in the isolated P. gingivalis and P. intermedia were compared using pulsed field gel electrophoresis (PFGE), the patterns in the natural teeth were in accordance with those in the implants in 3 of 4 cases (75.0%) in P. gingivalis and all cases in P. intermedia. This finding suggested that bacterial colonization around implants occurred early after the implant region was exposed to the intraoral cavity and that the bacteria were transmitted from the area around the natural teeth. PMID- 15536859 TI - A case of Antley-Bixler syndrome with severe skeletal Cl. III malocclusion. AB - Antley-Bixler syndrome is a disorder characterized by craniosynostosis, midface hypoplasia, choana blockade, and radiohumeral synostosis. However, the features of occlusion remain unclear. In this paper, we report a case of Antley-Bixler syndrome, a 7-year-old boy, from the viewpoint of orthodontics. From lateral cephalometric head film analysis, remarkable retardation of the anterior subcranial base, infraorbitale, and maxilla were notable, as was vertical growth restriction of the maxilla. The choana blockade tendency was also recognized. Moreover, although reverse occlusion was present, a mandibular retrognathic tendency was also present, and a short ramus mandible, remarkable mandibular vertical growth pattern, and skeletal open bite were present. In the dentition, two of the lower incisors were missing, and the present lower incisors were large. Maxillary and mandibular first molars were delayed in eruption. For treatment, the solutions to such remarkable skeletal problems were limited by the insufficiency of recovery of cranial formation after the operation. We planned a non-surgical treatment to expand the maxilla. It will be necessary to continually consider the treatment of his malocclusion as he continues to grow. PMID- 15536860 TI - Acinic cell carcinoma found by recurrence of a mucous cyst in the sublingual gland. AB - This case report describes an acinic cell carcinoma found by a recurrence of a ranula in the sublingual gland. A 42-year-old male was admitted to the hospital of the Tokyo Dental College with a swelling in his right oral floor but without pain. The lesion was treated by windowing the same day under the diagnosis of a ranula, but the swelling appeared again at the same area eight months after the first operation. A resection was performed, and the specimen was sent to the clinical laboratory for pathological diagnosis. Proliferating serous cells were seen in part of the wall of an exudative mucous cyst. PAS staining was partially positive, and immunohistochemical staining for S-100 protein, lactoferrin, and amylase were also positive in cytoplasmic granules. This report concludes that the pathological diagnosis is beneficial in clarifying the reasons for the recurrence of a benign lesion. PMID- 15536861 TI - Application of "prescale" as an aid to clinical diagnosis in orthodontics. AB - We examined the feasibility of applying the Dental Prescale Occluzer System to the evaluation of occlusion in orthodontic patients. The patients were classified according to the type of malocclusion or facial pattern. The values of occlusal contact area and occlusal force obtained from these individuals differed depending on the type of occlusion or facial pattern. These values were higher in individuals with normal occlusion than in patients with malocclusion and higher in patients with the Brachyo facial pattern than in those with the Dolico facial pattern. Because this system can very easily analyze the occlusal contact area, the mean pressure on each occlusal contact point, and the occlusal force during the process of orthodontic treatments, it can be useful as an aid for the initial diagnosis of occlusal problems, the observation of the changes in occlusion during treatment, and the establishment of the final goal of orthodontic treatments. PMID- 15536862 TI - On the use of human tissue after death. PMID- 15536863 TI - Supporting physicians through standards for appraisal and revalidation. PMID- 15536864 TI - Nutrition in health and disease. PMID- 15536865 TI - Nutrition and common health problems. PMID- 15536866 TI - Nutrition in acute care. PMID- 15536867 TI - Nutrition and mental health. PMID- 15536868 TI - Nutrition and older people: special considerations relating to nutrition and ageing. PMID- 15536869 TI - The slow death of the clinical post-mortem examination: implications for clinical audit, diagnostics and medical education. AB - The adult clinical post-mortem examination has seriously declined in Norwich recently, with only 34 of them (representing 1.4% of deaths in hospital) having been undertaken in 2003. Moreover, the next-of-kin are increasingly restricting the extent of the examination when they give consent. Analogous but less severe changes have occurred in the post-mortem examination of stillbirths and perinates. Many clinicians are unaware of these events, which may come to have wide-ranging detrimental effects. One possible cause is the lack of training of junior medical staff in obtaining consent for post-mortem examination, though other factors are also important. PMID- 15536870 TI - Influence of guidelines on CPR decisions: an audit of clerking proforma. AB - We audited documentation rates and implementation of cardiopulmonary resuscitation (CPR) decisions for patients admitted under the Department of Elderly Care Medicine, Mayday University Hospital, Croydon, as new guidelines and a proforma were introduced. For the first audit, data were collected from 75 departmental discharges. Following introduction of a proforma, six point prevalence audits were performed of all elderly care inpatients. Consultant documentation improved from 27/75 (36%) to 102/109 (94%), 135/148 (91%), 133/140 (95%), 96/119 (81%), 148/157 (94%) and 167/169 (98%) in audits 2, 3, 4, 5, 6 and 7 respectively. The percentages of decisions that were Do Not Attempt Resuscitation (DNAR) were 64% 72%, 45%, 68% and 62% in audits 3 to 7 respectively. For audit 5 our guidelines required discussion with patient before making a DNAR order, whereas the guidelines applicable for the other audits did not stipulate discussion. The fall in documentation rates and proportion of CPR decisions that were DNAR in audit 5 were statistically significant. There was no significant difference in age, diagnosis, cognitive function or disability between patients in those audits (3-7) when these parameters were recorded. Introducing a proforma significantly improved CPR decision documentation. Obligatory discussion with a patient before issuing a DNAR order was associated with a fall in documentation of decisions. PMID- 15536871 TI - Achieving compliance with the European Working Time Directive in a large teaching hospital: a strategic approach. AB - This paper describes the strategy which achieved European Working Time Directive (EWTD) compliance at the Royal Free Hampstead NHS Trust in medicine and surgery. Compliance with EWTD regulations was assessed by diary card exercise, clinical care assessed through critical incident reports, electronic handover documents and nursing reports, training opportunities assessed by unit training directors, cost controls assessed by finance department analysis, and workload assessed by staff attendance on wards, in casualty and in theatres. There was a change in focus of care to a consultant-led, specialist registrar- (SpR-)driven service extending into evenings and on weekends, coupled with a move to a multi-skilled team for night cover, and to a move from traditional on-call shifts to a full shift system across both medicine and surgery. Compliance with the EWTD was achieved whilst maintaining good standards of clinical care, ensuring training opportunities for doctors in training, controlling payroll costs, removing the need for locums, and reducing workload for both junior doctors and consultants. PMID- 15536872 TI - The prevention and treatment of glucocorticoid-induced osteoporosis in clinical practice. AB - Glucocorticoids are associated with increased risk of bone loss and fracture. This study compared the prescribing of bone protective agents by rheumatologists in clinical practice with the standards recommended in the 1998 UK Consensus guidelines. All glucocorticoid users who attended rheumatology outpatients during a four-week period were eligible. Notes were audited according to a predefined proforma. Among the 1290 rheumatology outpatients seen in the study period, 189 (15%) were taking glucocorticoids. 63% of glucocorticoid patients were taking calcium and 46% vitamin D. In total, 124 (71%) of the 175 patients available for review were at high risk of osteoporotic fracture, of whom 76 (61%) were taking appropriate prophylaxis. In 26 (15%) patients, insufficient information was available to be able to quantify the risk of fracture. The study showed that the audit standard was not met in 39% of cases. A better strategy for the monitoring of clinical risk factors is therefore required. PMID- 15536873 TI - Growth hormone--from molecule to mortality. AB - Growth hormone (GH) acts predominantly via insulin-like growth factor-I (IGF-I) expression. Acromegaly is associated with an increased mortality which can be reversed by optimal treatment. Somatostatin analogues are effective adjunctive therapies in patients treated with surgery and/or radiotherapy and result in tumour shrinkage in many patients. Pegvisomant is a GH analogue which inhibits functional dimerization of GH receptors, inhibits GH activity and normalizes IGF I in over 90% of subjects. Adult GH deficiency is associated with changes in body composition, insulin status, lipid profile and Quality of Life measures. Hypopituitarism is associated with an increased mortality. Replacement with GH has clinically beneficial effects but there are no data on effects on mortality. Taller individuals are at a 20-60 percent increased risk of a range of cancers, an effect that may be mediated via IGFs. These observations suggest that there is an optimal level of circulating GH and IGF-I required to maintain normal health. PMID- 15536874 TI - Preventing type 1 diabetes mellitus: hype and hope. AB - Autoimmune diseases affect 10% or more of the UK population. In organ-specific autoimmune diseases a particular tissue is targeted by the aggressive immune response. Type 1 diabetes is due to destruction of insulin-secreting islet cells. Both genetic and environmental factors cause type 1 diabetes and environmental events can operate very early, even in utero. The early induction of diabetes associated autoantibodies and the long prediabetic period means that autoimmune diabetes in children can be predicted by detecting these autoantibodies. The purpose of prediction is prevention and initial studies suggest that it is possible to modify the disease process. However, the impact of such therapy on the disease is as yet extremely modest. PMID- 15536875 TI - El Medico (the doctor). PMID- 15536876 TI - Chronic peripheral oedema: the critical role of the lymphatic system. PMID- 15536877 TI - Excessive daytime sleepiness and driving: regulations for road safety. AB - Individuals who fall asleep at the wheel usually do so because they are sleep deprived. It is likely that they are aware of the circumstances leading to sleepiness and of feeling sleepy before the event. Nevertheless, sleepiness sufficient to cause or contribute to an accident may involve a disorder of sleep, and little attention has been given to such disorders in the consideration of accident prevention. In this context, the Department for Transport brought together a group to explore the potential significance of sleep disorders in accidents. The Driver and Vehicle Licensing Agency has clarified existing regulations, particularly those that concern vocational drivers. PMID- 15536878 TI - The Renal National Service Framework: a step in the right direction. AB - Part one of the National Service Framework (NSF) for Renal Services was published early in 2004. The document covers the treatment of patients with end stage renal failure with dialysis and transplantation. Five standards to be achieved by 2014 are proposed. These cover access to information, timely preparation for dialysis, quality planned access surgery, patient choice for dialysis modality, and improvements in the quantity and success of renal transplants. These standards are underpinned by five early actions to be achieved by 2006. These include improved information flow about current practice and outcomes, increases in haemodialysis capacity and adherence to National Institute for Clinical Excellence guidelines on immunosuppression. The Renal NSF is welcome as a significant boost to the profile of renal services in England. Implementation will be difficult and will require the sustained pressure of the 'Kidney Alliance' representing patients, clinicians and supporting organisations. PMID- 15536879 TI - Academy of Medical Sciences: promoting advances in health science and biomedical research. AB - The Academy of Medical Sciences has matured quickly and found a distinctive niche amongst leading opinion formers and policy makers in healthcare and biomedical research. The Academy's 800 Fellows are the UK's leading medical scientists from hospitals, academia, public service and industry, and it is their expertise that gives the Academy its authority. The Academy campaigns vigorously to put science at the heart of the UK's public policy agenda in health, and to ensure that rapid advances in knowledge are translated as quickly as possible into benefits for patients. This paper briefly describes the work of the Academy and highlights the key achievements of the early years, particularly the Academy's role in drawing Government attention to the plight of academic medicine. The failure of clinical research to keep pace with scientific advances is now fully recognised and steps are being taken to establish new structures and to rebuild capacity in the UK. PMID- 15536880 TI - Human remains: objects to study or ancestors to bury? PMID- 15536881 TI - Maintaining a professional approach to life. PMID- 15536882 TI - Combination long-acting beta2-agonists and inhaled corticosteroids in chronic obstructive pulmonary disease. PMID- 15536883 TI - An audit into consultation in 'do not attempt resuscitation' decisions. PMID- 15536884 TI - Innocent beyond reasonable doubt? PMID- 15536885 TI - Effect of shift work on mental state of factory workers. AB - This paper examines the effects of shift work on the mental state of factory workers. As an indicator of the workers' mental condition, the authors used a scoring system (referred to below as the 'depression tendency score') based on the SRQ-D investigative report. The depression tendency score of the men was higher among the shift worker group than among the regular day worker group (p < 0.01). The depression tendency score of the male back-to-back shift workers was higher than that of the male regular day workers among skilled workers (p < 0.05). Among the women, no notable difference in depression tendency score was observed between the regular day worker group and the shift worker group. However, the depression tendency score of the female two-shift workers was higher than that of the female regular day workers among skilled workers (p < 0.05). We conclude that the mental health of men is easily affected by back-to-back shift work and that of women is affected by two-shift work because of the difference in modern societal/home role between man and woman. PMID- 15536886 TI - Anatomic and clinical investigation of a low signal peripheral line (black line) around the lumbar herniated nucleus pulposus on magnetic resonance imaging. AB - It has been reported that a low signal peripheral line (black line) around the lumbar herniated nucleus pulposus (HNP) on magnetic resonance imaging (MRI) can be used for the evaluation of the interruption of the posterior longitudinal ligament (PLL). However, the previous reports have showed that different rates of agreement between MRI and surgical findings. In order to clarify this matter, the black line on MRI was assessed by a combined anatomic and clinical investigation. Three cadavers were used for the anatomic part of the study. For the clinical study, 11 patients with the diagnosis as HNP and 5 healthy volunteers were subjected to MRI to compare with the results from the anatomic study. The lumbo sacral spine was dissected en bloc from the cadavers. The first imaging on MRI of the specimens was performed with the dural sac; the second imaging was performed after the dural sac and the nerve roots have been removed but with the PLL left; the third imaging was performed after the PLL was completely removed. After completion of imaging, the specimens were cut in sagittal and horizontal planes for histological evaluation. In the cadavers after removing the PLL histologically, the black lines were still shown on MRI. Finally, after changing encoding, the black lines were interrupted at some disc levels in the cadaver specimen, the patients with HNP, and healthy volunteers. Therefore the black lines could be interpreted as a chemical shift artifact. These results indicate that the continuity or the discontinuity of the black line is not appropriate sign to diagnose whether disruption of the PLL has occurred or not. PMID- 15536887 TI - Polymyositis associated with urinary bladder cancer: an autopsy case. AB - A 56-year-old man suffered from muscle weakness with elevated serum creatine kinase. Under diagnosis of polymyositis, the patient was treated with corticosteroid, methotrexate and cyclosporin A. Eleven months after the first signs of muscle weakness, the patient suffered an abrupt onset of anuria and underwent hemodialysis. The patient died of respiratory insufficiency 14 months after the first signs of muscle weakness. Autopsy findings revealed associated urinary bladder cancer with histological indications of adenosquamous cell carcinoma, liver metastasis and cancerous lymphangitis of the lung. PMID- 15536888 TI - A surgical case of aortic valve replacement in patient with chronic idiopathic thrombocytopenic purpura. AB - A 64-year-old man was presented to another hospital with bleeding from the upper jaw with a platelet count of 0.1 x 10(4)/microl one year ago, and idiopathic thrombocytopenic purpura complicated with aortic stenosis and regurgitation was diagnosed. Corticosteroid administration was initiated and the patient was admitted to our hospital for surgery two months after confirmation of symptoms associated with cerebral ischemia. Corticosteroid was administered for one year until the day of surgery, and aortic valve replacement was performed with a platelet count of 8.4 x 10(4)/microl. During surgery, bleeding tendency with decreased platelets was confirmed, which was corrected with intraoperative platelet transfusion, postoperative gamma-globulin administration, and postoperative oral corticosteroid administration. Caution must be exercised against perioperative bleeding tendency in open heart surgery, even when platelet count is maintained using small amounts of preoperative corticosteroid. PMID- 15536889 TI - Target controlled infusions: targeting the effect site while limiting peak plasma concentration. AB - BACKGROUND: Target controlled infusion (TCI) systems are available commercially worldwide, except in North America. Existing systems target the plasma drug concentration, which may be less than ideal because the plasma is not the site of drug effect. The commonly accepted existing algorithm to target the site of drug effect results in high plasma concentrations, which may increase the acute hemodynamic effects of intravenous anesthetic drugs. METHODS: A novel mathematical algorithm is proposed for controlling the effect site concentration using a TCI device. The algorithm limits the peak plasma concentration, thereby slowing the onset of anesthetic drug effect but potentially ameliorating side effects. Simulations are used to examine the delay in time to peak effect for fentanyl, alfentanil, sufentanil, remifentanil, and propofol when the peak plasma concentration is limited by the algorithm. RESULTS: The plasma overshoot required of the previously proposed algorithm to control the site of drug effect can be reduced by 60% with only about a 20% delay in the onset of drug effect. CONCLUSIONS: Concerns about the high plasma concentrations that result from targeting the effect site with a TCI device can be addressed by reducing the peak plasma concentration by as much as 60% with only a very modest increase in time to peak effect. PMID- 15536890 TI - On the use of multivariable piecewise-linear models for predicting human response to anesthesia. AB - The standard modeling paradigm used to describe the relationship between input anesthetic agents and output patient endpoint variables are single-input single output pharmacokinetic-pharmacodynamic (PK-PD) compartment models. In this paper, we propose the use of multivariable piecewise-linear models to describe the relations between inputs that include anesthesia, surgical stimuli and disturbances to a variety of patient output variables. Subspace identification methods are applied to clinical data to construct the models. A comparison of predicted and measured responses is completed, which includes predictions from PK PD models, and piecewise-linear time-invariant models. PMID- 15536891 TI - A mathematical model for breast cancer lesion estimation: electrical impedance technique using TS2000 commercial system. AB - We present a mathematical model to analyze transadmittance data for the detection of breast cancer using TransScan TS2000 commercial system. The model was constructed based on the assumption that a lesion exists near the surface of a breast region. The breast region that is considered as a background is assumed to be homogeneous at least near the surface where we attach a planar array of electrodes. Based on the model, we developed a lesion estimation algorithm utilizing single- or multifrequency transadmittance data. The approximate ratio of two conductivity values for the lesion and background needs to be known to estimate the size of the lesion even though the location estimate does not require this ratio. From the results of numerical simulations with added noise, we suggest better ways of interpreting TS2000 transadmittance images for the detection of breast cancer with improved accuracy. Since this study provides a rigorous mathematical modeling of TS2000 commercial system, it will be possible to apply the technique to lesion estimation problems based on more realistic models of breast regions in future studies. PMID- 15536892 TI - Peripheral nerve stimulation by gradient switching fields in magnetic resonance imaging. AB - A heterogeneous model of the human body and the scalar potential finite difference method are used to compute electric fields induced in tissue by magnetic field exposures. Two types of coils are considered that simulate exposure to gradient switching fields during magnetic resonance imaging (MRI). These coils producing coronal (y axis) and axial (z axis) magnetic fields have previously been used in experiments with humans. The computed fields can, therefore, be directly compared to human response data. The computed electric fields in subcutaneous fat and skin corresponding to peripheral nerve stimulation (PNS) thresholds in humans in simulated MRI experiments range from 3.8 to 5.8 V/m for the fields exceeded in 0.5% of tissue volume (skin and fat of the torso). The threshold depends on coil type and position along the body, and on the anatomy and resolution of the human body model. The computed values are in agreement with previously established thresholds for neural stimulation. PMID- 15536893 TI - A theoretical model of the high-frequency arrhythmogenic depolarization signal following myocardial infarction. AB - Theoretical body-surface potentials were computed from single, branching and tortuous strands of Luo-Rudy dynamic model cells, representing different areas of an infarct scar. When action potential (AP) propagation either in longitudinal or transverse direction was slow (3-12 cm/s), the depolarization signals contained high-frequency (100-300 Hz) oscillations. The frequencies were related to macroscopic propagation velocity and strand architecture by simple formulas. Next, we extended a mathematical model of the QRS-complex presented in our earlier work to simulate unstable activation wavefront. It combines signals from different strands with small timing fluctuations relative to a large repetitive QRS-like waveform and can account for dynamic changes of real arrhythmogenic micropotentials. Variance spectrum of wavelet coefficients calculated from the composite QRS-complex contained the high frequencies of the individual abnormal signals. We conclude that slow AP propagation through fibrotic regions after myocardial infarction is a source of high-frequency arrhythmogenic components that increase beat-to-beat variability of the QRS, and wavelet variance parameters can be used for ventricular tachycardia risk assessment. PMID- 15536894 TI - Extraction and quantification of left ventricular deformation modes. AB - We have developed a method that decomposes the deformation of the left ventricle (LV) between end diastole (ED) and end systole (ES) into separate deformation modes such as longitudinal shortening, wall thickening, and twisting. The deformation was initially found from the motion of an LV finite-element mesh that was fitted to clinically obtained magnetic resonance (MR) tagged images. A mode coefficient was calculated for each deformation mode to quantify the different modes and, thus allowing for discrimination of normal and abnormal deformation patterns. We applied the method to 13 normal subjects and 13 diabetes patients. By using the ED mesh as reference and adding the extracted deformation modes multiplied by their mode coefficients, an approximate ES mesh was calculated and compared with the "true" ES mesh found from the MR images. For the 26 subjects the average Euclidean distance was less than 1.7+/-0.9 mm between the nodes of the approximated and true ES meshes. The coefficient values for the patient group showed significantly less longitudinal shortening, less wall thickening, more longitudinal twisting and also more bulging of the septum into the LV when compared with the normal subjects. We conclude that the developed method successfully quantifies the deformation into several modes of deformation and is capable of distinguishing the deformation of a group of patients from a group of normal subjects. PMID- 15536895 TI - Nonlinear modeling of the dynamic effects of arterial pressure and CO2 variations on cerebral blood flow in healthy humans. AB - The effect of spontaneous beat-to-beat mean arterial blood pressure fluctuations and breath-to-breath end-tidal CO2 fluctuations on beat-to-beat cerebral blood flow velocity variations is studied using the Laguerre-Volterra network methodology for multiple-input nonlinear systems. The observations made from experimental measurements from ten healthy human subjects reveal that, whereas pressure fluctuations explain most of the high-frequency blood flow velocity variations (above 0.04 Hz), end-tidal CO2 fluctuations as well as nonlinear interactions between pressure and CO2 have a considerable effect in the lower frequencies (below 0.04 Hz). They also indicate that cerebral autoregulation is strongly nonlinear and dynamic (frequency-dependent). Nonlinearities are mainly active in the low-frequency range (below 0.04 Hz) and are more prominent in the dynamics of the end-tidal CO2-blood flow velocity relationship. Significant nonstationarities are also revealed by the obtained models, with greater variability evident for the effects of CO2 on blood flow velocity dynamics. PMID- 15536896 TI - Adaptive denoising of event-related functional magnetic resonance imaging data using spectral subtraction. AB - A new adaptive signal-preserving technique for noise suppression in event-related functional magnetic resonance imaging (fMRI) data is proposed based on spectral subtraction. The proposed technique estimates a parametric model for the power spectrum of random noise from the acquired data based on the characteristics of the Rician statistical model. This model is subsequently used to estimate a noise suppressed power spectrum for any given pixel time course by simple subtraction of power spectra. The new technique is tested using computer simulations and real data from event-related fMRI experiments. The results show the potential of the new technique in suppressing noise while preserving the other deterministic components in the signal. Moreover, we demonstrate that further analysis using principal component analysis and independent component analysis shows a significant improvement in both convergence and clarity of results when the new technique is used. Given its simple form, the new method does not change the statistical characteristics of the signal or cause correlated noise to be present in the processed signal. This suggests the value of the new technique as a useful preprocessing step for fMRI data analysis. PMID- 15536897 TI - The effects of maturation on early and late phases of phrenic neurogram during eupnea. AB - In this paper, we investigate the effects of maturation on the early and late phases of the phrenic neurogram. We have used the matching pursuit (MP) method to examine the effects of maturation on breathing patterns in both time and frequency domains. The MP was chosen since the wavelet transform method may not represent signals whose Fourier transforms have a narrow high-frequency support. The phrenic neurogram was recorded from 25 piglets (3-35 days) during eupnea (normal breathing) at three postnatal age groups: young (3-7 days (n = 9)), (middle) 10-21 days (n = 6), and old (29-35 days (n = 10)). The energy percentage of atoms representing the nonperiodic neural activities (NPNAs) significantly decreased from young age to middle age groups (p<0.01) and from young age to old age groups (p<0.01), and from middle age to old age groups (p<0.055) in the early phase (the first half) of the phrenic neurogram, but these changes were not statistically significant in the late phase (the second half) of the phrenic neurogram as maturation proceeded. However, the energy percentage of atoms representing the periodic neural activities (PNAs) decreased with maturation, but these changes were not statistically significant in the early phase of the phrenic neurogram. The energy percentage of (PNAs) increased in the late phase of the phrenic neurogram as maturation proceeded although these changes were only significant between young and old age groups (p<0.01). These results suggest that the significant decrease of the NPNAs in the early phase and the increase in the late phase of the phrenic neurogram could be a sign of maturation in piglets. PMID- 15536898 TI - Parallel multigrid preconditioner for the cardiac bidomain model. AB - The bidomain equations are widely used for the simulation of electrical activity in cardiac tissue but are computationally expensive, limiting the size of the problem which can be modeled. The purpose of this study is to determine more efficient ways to solve the elliptic portion of the bidomain equations, the most computationally expensive part of the computation. Specifically, we assessed the performance of a parallel multigrid (MG) preconditioner for a conjugate gradient solver. We employed an operator splitting technique, dividing the computation in a parabolic equation, an elliptical equation, and a nonlinear system of ordinary differential equations at each time step. The elliptic equation was solved by the preconditioned conjugate gradient method, and the traditional block incomplete LU parallel preconditioner (ILU) was compared to MG. Execution time was minimized for each preconditioner by adjusting the fill-in factor for ILU, and by choosing the optimal number of levels for MG. The parallel implementation was based on the PETSc library and we report results for up to 16 nodes on a distributed cluster, for two and three dimensional simulations. A direct solver was also available to compare results for single processor runs. MG was found to solve the system in one third of the time required by ILU but required about 40% more memory. Thus, MG offered an attractive tradeoff between memory usage and speed, since its performance lay between those of the classic iterative methods (slow and low memory consumption) and direct methods (fast and high memory consumption). Results suggest the MG preconditioner is well suited for quickly and accurately solving the bidomain equations. PMID- 15536899 TI - Analyzing the dynamic wavefront aberrations in the human eye. AB - The optics of the human eye are not static in steady viewing conditions and exhibit microfluctuations. Previous methods used for analyzing dynamic changes in the eye's optics include simple Fourier-transform-based methods, which have been used in studies of the eye's accommodation response. However, dedicated tools for the analysis of dynamic wavefront aberrations have not been reported. We propose a set of signal processing tools, the combination of which uncovers aspects of the dynamics of eye's optical aberrations which were hidden from conventional analysis techniques. The methodology includes extraction of artifacts from potentially significant eye movements, filtering, optimal parametric signal modeling, and frequency and time-frequency representations. The exposition of the techniques and their advantages over traditional techniques is illustrated for real dynamic eye wavefront aberration measurements. PMID- 15536900 TI - Maximum-likelihood estimation of low-rank signals for multiepoch MEG/EEG analysis. AB - A maximum-likelihood-based algorithm is presented for reducing the effects of spatially colored noise in evoked response magneto- and electro-encephalography data. The repeated component of the data, or signal of interest, is modeled as the mean, while the noise is modeled as the Kronecker product of a spatial and a temporal covariance matrix. The temporal covariance matrix is assumed known or estimated prior to the application of the algorithm. The spatial covariance structure is estimated as part of the maximum-likelihood procedure. The mean matrix representing the signal of interest is assumed to be low-rank due to the temporal and spatial structure of the data. The maximum-likelihood estimates of the components of the low-rank signal structure are derived in order to estimate the signal component. The relationship between this approach and principal component analysis (PCA) is explored. In contrast to prestimulus-based whitening followed by PCA, the maximum-likelihood approach does not require signal-free data for noise whitening. Consequently, the maximum-likelihood approach is much more effective with nonstationary noise and produces better quality whitening for a given data record length. The efficacy of this approach is demonstrated using simulated and real MEG data. PMID- 15536901 TI - Registration and real-time visualization of transcranial magnetic stimulation with 3-D MR images. AB - This paper describes a method for registering and visualizing in real-time the results of transcranial magnetic stimulations (TMS) in physical space on the corresponding anatomical locations in MR images of the brain. The method proceeds in three main steps. First, the patient scalp is digitized in physical space with a magnetic-field digitizer, following a specific digitization pattern. Second, a registration process minimizes the mean square distance between those points and a segmented scalp surface extracted from the magnetic resonance image. Following this registration, the physician can follow the change in coil position in real time through the visualization interface and adjust the coil position to the desired anatomical location. Third, amplitude of motor evoked potentials can be projected onto the segmented brain in order to create functional brain maps. The registration has subpixel accuracy in a study with simulated data, while we obtain a point to surface root-mean-square error of 1.17+/-0.38 mm in a 24 subject study. PMID- 15536902 TI - Object detection by correlation coefficients using azimuthally averaged reference projections. AB - A method of computing correlation coefficients for object detection that takes advantage of using azimuthally averaged reference projections is described and compared with two alternative methods-computing a cross-correlation function or a local correlation coefficient versus the azimuthally averaged reference projections. Two examples of an application from structural biology involving the detection of projection views of biological macromolecules in electron micrographs are discussed. It is found that a novel approach to computing a local correlation coefficient versus azimuthally averaged reference projections, using a rotational correlation coefficient, outperforms using a cross-correlation function and a local correlation coefficient in object detection from simulated images with a range of levels of simulated additive noise. The three approaches perform similarly in detecting macromolecular views in electron microscope images of a globular macrolecular complex (the ribosome). The rotational correlation coefficient outperforms the other methods in detection of keyhole limpet hemocyanin macromolecular views in electron micrographs. PMID- 15536903 TI - Image enhancement in the JPEG domain for people with vision impairment. AB - An image enhancement algorithm for low-vision patients was developed for images compressed using the JPEG standard. The proposed algorithm enhances the images in the discrete cosine transform domain by weighting the quantization table in the decoder. Our specific implementation increases the contrast at all bands of frequencies by an equal factor. The enhancement algorithm has four advantages: 1) low computational cost; 2) suitability for real-time application; 3) ease of adjustment by end-users (for example, adjusting a single parameter); and 4) less severe block artifacts as compared with conventional (post compression) enhancements. Experiments with visually impaired patients show improved perceived image quality at moderate levels of enhancement but rejection of artifacts caused by higher levels of enhancement. PMID- 15536904 TI - Performance of large-size superconducting coil in 0.21T MRI system. AB - A high-temperature superconductor (HTS) was used on magnetic resonance imaging (MRI) receiver coils to improve image quality because of its intrinsic low electrical resistivity. Typical HTS coils are surface coils made of HTS thin-film wafers. Their applications are severely limited by the field of view (FOV) of the surface coil configuration, and the improvement in image quality by HTS coil is also reduced as the ratio of sample noise to coil noise increases. Therefore, previous HTS coils are usually used to image small in vitro samples, small animals, or peripheral human anatomies. We used large-size HTS coils (2.5-, 3.5-, and 5.5-in mean diameter) to enhance the FOV and we evaluated their performance through phantom and human MR images. Comparisons were made among HTS surface coils, copper surface coils, and cool copper surface coils in terms of the signal to-noise ratio (SNR) and sensitivity profile of the images. A theoretical model prediction was also used to compare against the experimental result. We then selected several human body parts, including the wrist, feet, and head, to illustrate the advantage of HTS coil over copper coil when used in human imaging. The results show an SNR gain of 200% for 5.5-in HTS coil versus same size copper coils, while for 2.5- and 3.5-in coils it is 250%. We also address the various factors that affect the performance of large size HTS coils, including the coil to-sample spacing due to cryogenic probe and the coil-loading effect. PMID- 15536905 TI - Enhancing interference rejection of preamplified electrodes by automated gain adaption. AB - The use of preamplified electrodes in electrocardiography (ECG) or electroencephalography (EEG) has a numbe of benefits, most importantly, the noise reduction through preamplification and the interference reduction through buffering (Fernandez and Pallas-Areny, 1997). Normally, preamplified electrodes are not used because the mismatch in amplification due to component tolerances leads to a reduced common mode rejection ration (CMRR) (Pallas-Areny and Webster, 1991). In this paper, we introduce a new technique to restore the CMRR. We do this by adapting the gain of the differential amplifier (DA) following the preamplified electrodes. The technique is based on measuring the transfer function of a chosen common mode signal. The gain of the DA is adjusted by a closed loop controller to maximize the CMRR. With this technique, there is no adverse effect on the ECG or EEG. In this way, the control loop can be operated continuously if needed. PMID- 15536906 TI - Development and evaluation of a new body-seat interface shape measurement system. AB - A new system has been developed to capture the body-seat interface shape. It can repeatedly and accurately measure interface deformation. The shape sensing array system uses optical fiber technology and is noninvasive. The system can cover an interface as large as 400 x 480 mm and the shape is measured over a 10 x 12 array of sensors laminated on ribbon substrates. The accuracy and repeatability of this system were assessed. Measurement errors were evaluated by comparing the shape with a reference shape obtained by a mechanical digitizer. The root-mean-square error in the Z direction for the system was 3.79 mm. The repeatability of the system was within 0.38 mm under controlled conditions. Different interface materials noticeably affected measurements. With the development of this interface shape measurement device, the basic information gathered through its use may prove to be fundamental in the successful design of generic-shape contoured support surfaces. Furthermore, we expect that the new shape measurement device will provide a quick and effective tool for cushion evaluation and clinical guidelines for cushion prescription. PMID- 15536907 TI - Long-term characterization of firing dynamics of spontaneous bursts in cultured neural networks. AB - Extracellular action potentials were recorded from developing dissociated rat neocortical networks continuously for up to 49 days in vitro using planar multielectrode arrays. Spontaneous neuronal activity emerged toward the end of the first week in vitro and from then on exhibited periods of elevated firing rates, lasting for a few days up to weeks, which were largely uncorrelated among different recording sites. On a time scale of seconds to minutes, network activity typically displayed an ongoing repetition of distinctive firing patterns, including short episodes of synchronous firing at many sites (network bursts). Network bursts were highly variable in their individual spatio-temporal firing patterns but showed a remarkably stable underlying probabilistic structure (obtained by summing consecutive bursts) on a time scale of hours. On still longer time scales, network bursts evolved gradually, with a significant broadening (to about 2 s) in the third week in vitro, followed by a drastic shortening after about one month in vitro. Bursts at this age were characterized by highly synchronized onsets reaching peak firing levels within less than ca. 60 ms. This pattern persisted for the rest of the culture period. Throughout the recording period, active sites showed highly persistent temporal relationships within network bursts. These longitudinal recordings of network firing have, thus, brought to light a reproducible pattern of complex changes in spontaneous firing dynamics of bursts during the development of isolated cortical neurons into synaptically interconnected networks. PMID- 15536908 TI - Model neural prostheses with integrated microfluidics: a potential intervention strategy for controlling reactive cell and tissue responses. AB - Model silicon intracortical probes with microfluidic channels were fabricated and tested to examine the feasibility of using diffusion-mediated delivery to deliver therapeutic agents into the volume of tissue exhibiting reactive responses to implanted devices. Three-dimensional probe structures with microfluidic channels were fabricated using surface micromachining and deep reactive ion etching (DRIE) techniques. In vitro functional tests of devices were performed using fluorescence microscopy to record the transient release of Texas Red labeled transferrin (TR-transferrin) and dextran (TR-dextran) from the microchannels into 1% w/v agarose gel. In vivo performance was characterized by inserting devices loaded with TR-transferrin into the premotor cortex of adult male rats. Brain sections were imaged using confocal microscopy. Diffusion of TR-transferrin into the extracellular space and uptake by cells up to 400 microm from the implantation site was observed in brain slices taken 1 h postinsertion. The reactive tissue volume, as indicated by the presence of phosphorylated mitogen activated protein kinases (MAPKs), was characterized using immunohistochemistry and confocal microscopy. The reactive tissue volume extended 600, 800, and 400 microm radially from the implantation site at 1 h, 24 h, and 6 weeks following insertion, respectively. These results indicate that diffusion-mediated delivery can be part of an effective intervention strategy for the treatment of reactive tissue responses around chronically implanted intracortical probes. PMID- 15536909 TI - A laser speckle imaging technique for measuring tissue perfusion. AB - Laser Doppler imaging (LDI) has become a standard method for optical measurement of tissue perfusion, but is limited by low resolution and long measurement times. We have developed an analysis technique based on a laser speckle imaging method that generates rapid, high-resolution perfusion images. We have called it laser speckle perfusion imaging (LSPI). This paper investigates LSPI output and compares it to LDI using blood flow models designed to simulate human skin at various levels of pigmentation. Results show that LSPI parameters can be chosen such that the instrumentation exhibits a similar response to changes in red blood cell concentration (0.1%-5%, 200 microL/min) and velocity (0-800 microL/min, 1% concentration) and, given its higher resolution and quicker response time, could provide a significant advantage over LDI for some applications. Differences were observed in the LDI and LSPI response to tissue optical properties. LDI perfusion values increased with increasing tissue absorption, while LSPI perfusion values showed a slight decrease. This dependence is predictable, owing to the perfusion algorithms specific to each instrument, and, if properly compensated for, should not influence each instrument's ability to measure relative changes in tissue perfusion. PMID- 15536910 TI - A mathematical basis for the application of the modified geometric method to maximum frequency estimation. AB - The application of ultrasound in assessing the fetal cardiovascular system often requires the accurate estimation of maximum blood flow velocity waveforms using Doppler measurements. The modified geometric method estimates the maximum Doppler frequency as the frequency at which the vertical distance between the integrated spectrum and the reference line that connects the origin to the maximum value of the integrated spectrum is the largest. This paper presents a mathematical formulation for a class of maximum blood flow velocity estimation algorithms that includes the modified geometric method. The analysis provides a rationale for the continued use of the modified geometric method for estimating the maximum frequency envelopes of Doppler signals. This paper also contains experimental results demonstrating the superiority of the modified geometric method over a commonly used threshold crossing method. PMID- 15536911 TI - Prescriptions sans frontieres (or how I stopped worrying about Viagra on the Web but grew concerned about the future of healthcare delivery). PMID- 15536912 TI - Genetic diagnosis and intellectual property rights: a proposal to amend "the physician immunity statute". PMID- 15536913 TI - Cloning matters: how Lawrence v. Texas protects therapeutic research. PMID- 15536914 TI - Putting international research ethics guidelines to work for the benefit of developing countries. PMID- 15536915 TI - How do international trade agreements influence the promotion of public health?- An introduction to the issue. PMID- 15536916 TI - The "good old days" of TRIPS: the U.S. trade agenda and the extension of pharmaceutical test data protection. PMID- 15536917 TI - The interaction of increased trade and the decentralization of health care delivery in Nepal: a suggestion for reform. PMID- 15536918 TI - Cambodia's membership in the WTO and the implications for public health. PMID- 15536919 TI - Barrier to trade or barrier to profit? Why Australia's Pharmaceutical Benefits Scheme worries U.S. drug companies. PMID- 15536920 TI - Australia-United States Free Trade Agreement and the Australian Pharmaceutical Benefits Scheme. PMID- 15536921 TI - Trade agreements, intellectual property, and the role of the World Bank in improving access to medicines in developing countries. PMID- 15536922 TI - Born too soon. PMID- 15536923 TI - How to prepare for a winter season of vaccine shortage. PMID- 15536924 TI - Drugs in labor. A new hope. PMID- 15536925 TI - Marion's message. Another good reason to prevent cesarean section. PMID- 15536926 TI - How do we educate women about the use and effects of drugs in labor? PMID- 15536927 TI - Adverse events following misoprostol induction of labor. PMID- 15536928 TI - Drugs in labor: an overview. PMID- 15536929 TI - Knitting midwives for drugless childbirth? PMID- 15536930 TI - Evidence supports the obvious. PMID- 15536931 TI - HIV & pregnancy--what more can we do for HIV-positive women? PMID- 15536932 TI - A determined life. PMID- 15536933 TI - Saving our endangered midwifery allies. PMID- 15536934 TI - The nesting instinct. PMID- 15536935 TI - Meet the practitioner. PMID- 15536936 TI - Preserving the breastfeeding relationship: hypoglycemia in the breastfed infant- fact or fiction? PMID- 15536937 TI - The past, present & future of midwifery. PMID- 15536938 TI - Did immunizations kill my babies? PMID- 15536939 TI - Compassionate C-section. PMID- 15536940 TI - The business of midwifery. Managed care--the philosophy. PMID- 15536941 TI - Maiden midwives. PMID- 15536942 TI - ICM and hemorrhage. PMID- 15536943 TI - Midwifery in Israel. AB - Socialized medicine provides free and easily accessible prenatal and intrapartum care in Israel. Mothers also receive the equivalent of dollars 100 to dollars 300 after delivering in hospital. The infant mortality rate is 5.5 per 1000 live births. Uncomplicated births (80%) are attended by midwives. Midwives routinely rupture membranes, attach electronic fetal monitoring, give intravenous fluids and deliver in lithotomy position. Approximately 15% of women undergo induction, 20% augmentation, 40% epidural, 30% episiotomy, 17% cesarean section. In 2003, 1% of births were cesareans by maternal request, rather than medical necessity, and 0.1% of births were planned homebirths. The system processes 140,000 births per year with good medical outcomes, especially considering Israel is only 56 years old. PMID- 15536944 TI - "There is a foot!". PMID- 15536945 TI - Placental rituals, placental medicine. PMID- 15536946 TI - Antonina Sanchez Mendez. PMID- 15536947 TI - Marti's homebirth in Hungary. PMID- 15536948 TI - [Mutational analysis of the vhl gene in the sporadic renal cell carcinoma]. AB - OBJECTIVES: To assess and characterize mutations in the vhl gene in tumor tissue samples from 30 patients undergoing surgery for renal cell carcinoma in our department. METHODS: Descriptive, observational and analytical study of 30 patients undergoing surgery for RCC, analyzing the vhl gene sequence in tumor tissue, and using healthy renal tissue from the same patients as controls. Tissues were processed by DNA extraction, PCR amplification of the three exons that conform the gene, and ulterior automatic sequence analysis of the amplified exons between intronic primers previously designed. The sequence is compared with the corresponding exons included in the GeneBank. Alterations were checked by backwards sequence analysis. RESULTS: 9 mutations (30%) were found in the tumoral samples analyzed. 7 of them were punctual (one of them intronic); the other two were deletions. Mutations were distributed among the three exons: 3 in exon 1, 4 in exon 2, 1 in exon 3 and 1 intronic. One of the samples showed 2 mutations. Control tissue was free of mutations. CONCLUSIONS: Sporadic RCC shows mutations in the vhl gene which mainly appear in the clear cell subtype. Such alterations result in severe disturbances in the protein, disturbing its tumor suppressing function. PMID- 15536949 TI - [Radical prostatectomy as monotherapy for locally advanced prostate cancer (T3a): 12 years follow-up]. AB - OBJECTIVES: To evaluate the oncological and functional results of radical prostatectomy as monotherapy for stage T3a prostate cancer. METHODS: We include our initial and consecutive series of 83 patients with prostate cancer (studied by digital rectal examination and transrectal ultrasound) who had not received neoadjuvant treatment undergoing radical prostatectomy from July 1988 to December 2003. No patient received adjuvant treatment, and deferred intermittent androgen blockade was used when patients with biochemical progression exceeded a PSA of 4ng/ml. Up RESULTS: After a mean follow-up of 68.7 (1-139) months: overall and specific survival 97.6% and 100% respectively; biochemical progression 36.1% (22 pT2 (0%), 41 p T3a (36.6%), 13 pT3b (61.5%) and 7pT4a (100%)). Positive margins 61.4% (41.2% unifocal with a progression rate of 23.8%). 96.4% achieved continence and 39.6% recovered potency. Among 30 patients with biochemical progression, 19 required treatment with deferred intermittent androgen blockade (one cycle in 10 patients, two cycles in six, and three cycles in the remaining three). CONCLUSIONS: Our results support the indication of radical prostatectomy as single therapy without neoadjuvant treatment as a curative indication for locally advanced prostate cancer (T3a) whenever complete excision is expected: Gleason 7 T3 tumors without diffuse extension on ultrasound. 26.5% of these T3a patients were overstaged and resulted to be organ-confined (pT2). Ten-year probability of biochemical progression-free survival was 100% for pT2 and 81.9% for the lower risk pT3a (well or moderately-differentiated with negative surgical margins or unifocal). Functional results for T3a were similar to the ones of the clinically-localized (T2) series for both retropubic and perineal approaches. 30 patients had developed biochemical progression at the time of study closure and were free of hormonal treatment during 81.6% of the total follow-up time with our deferred intermittent androgen blockade treatment line, so that we consider we can offer it as the first treatment option for progression providing a maximal quality of life and allowing ulterior second line therapies. Patients who mainly benefited were those on progression who have recovered sexual function: 41.7% of potent patients after radical prostatectomy recovered potency again over the second phase (no treatment) of the deferred intermittent androgen blockade. PMID- 15536950 TI - [Usefulness of bone-specific alkaline phosphatase for bone metastases detection in prostate cancer]. AB - OBJECTIVES: To assess the clinical usefulness of bone-specific alkaline phosphatase (B-AKP) to detect or rule out bone metastases in patients with prostate cancer. METHODS: Between November 2001 and July 2003 111 patients with the diagnosis of prostate adenocarcinoma were evaluated to rule out bone metastases. Bone metastases were studied after clinical suspicion, PSA value higher than 10 ng/ml, or Gleason score higher than 7. Bone scan was used as detection test. Patients were tested for serum B-AKP with a reference value equal or higher than 30ng/ml. The sensitivity, specificity, positive predictive value and negative predictive value of B-AKP were measured in comparison to bone scan as the gold standard. RESULTS: 111 patients with the diagnosis of prostate adenocarcinoma were evaluated to rule out bone metastases. Mean age was 69.70 years (50-83 years). Average B-AKP value was 29.28 ng/ml (9-56 ng/ml). Bone scan was positive in 31 patients. Among these 31, 26 patients had positive B-AKP and five negative. AP patients had negative bone scan. From this AP patients, B-AKP was positive in 17 and negative in 63. The sensitivity of the test was 83.8%. The specificity was 78%. Positive predictive value was 60% and negative predictive value was 92%. CONCLUSIONS: Bone alkaline phosphatase is a greatly useful test for the study of patients with prostate cancer in whom bone metastases are suspected either to confirm them or to rule out their presence. PMID- 15536951 TI - [Bladder augmentation and urinary diversion in kidney transplant candidates]. AB - OBJECTIVES: Kidney transplantation (KT) is the treatment of choice for patients with end stage renal disease (ESRD). 6% of the patients develop ESRD due to congenital or acquired lower urinary tract anomalies, which sometimes imply the need of an additional surgical procedure to make the patient suitable for transplantation. METHODS: We review 6 cases of KT receptors (three of them pediatric) to whom some kind of reconstruction of the lower urinary tract with bowel was performed over the last 10 years. RESULTS: Most frequent etiologies: neurogenic bladder (3), small and contracted bladder after genitourinary tuberculosis, urethral valves, and transitional cell carcinoma with radical cystoprostatectomy and bilateral nephrectomy. Surgical techniques: Bladder augmentation with colon (3), Bricker's defunctionalized ileal loop (2), and Goodwin 's ileal bladder augmentation; all of them were performed between 8 and 147 months before transplant. COMPLICATIONS: UTI in 2 patients. Recurrent stenosis of the ileal loop in one patient who required endoscopic balloon dilation and stent placement in the stenotic segment with poor results and finally requiring loop reconstruction. Another patient developed stenosis of the ureteroneocystostomy anastomosis and reimplant was performed. All of them had good outcome. One case had a subacute kidney rejection episode with good response to steroids. No graft was lost. Current serum creatinine values are between 0.69 and 2.6. CONCLUSIONS: The use of bowel in patients with pathologic bladders is as safe method to allow these patients to receive a kidney transplant when bladder rehabilitation has not been possible with conservative measures. PMID- 15536952 TI - [Urologic damage of the primary antiphospholipid syndrome]. AB - OBJECTIVES: To perform a review and update of the antiphospholipid syndrome summarizing its urological presentations. METHODS: A complete bibliographic search was performed through PubMed MEDLINE and articles were reviewed with special attention to those bibliographic references about urological presentations. We document the unique and unpublished case of a patient with neurogenic bladder secondary to antiphospholipid syndrome. RESULTS/CONCLUSIONS: The antiphospholipid syndrome is an acquired autoimmune systemic disease generating a permanent hypercoagulability status with recurrent multiorgan thrombotic events due to circulating antiphospholipid antibodies. It may be secondary to a heterogeneous group of diseases (mainly lupus) and drugs, or primary if it appears isolated without any demonstrable systemic disease or concomitant medication. It is mainly characterized by venous or arterial recurrent thrombosis, recurrent abortion, thrombocytopenia, and circulating antiphospholipid auto-antibodies. Treatment with anticoagulants and correction of the hypercoagulable status contributing factors, arterial or venous thrombosis, and vascular risk aim to avoid new thrombosis episodes. Genitourynary system may be affected in any of its parts, generally by arterial or venous thrombosis. Kidney is the most frequently affected organ, in addition to transplanted kidney grafts, adrenal glands, bladder and testicles. There is a relationship between antiphospholipid syndrome and infertility. For the first time, we describe bladder involvement presenting as hyperreflexic neurogenic bladder with detrusor sphincter dyssynergia after spontaneous spinal cord thrombosis in an asymptomatic adolescent with primary antiphospholipid syndrome which was unknown before. PMID- 15536953 TI - [Current indications of percutaneous nephrostomy associated with extracorporeal shock wave lithotripsy in the treatment of renoureteral stones]. AB - OBJECTIVES: To evaluate the usefulness of PN as urinary diversion in the treatment of urinary stones by ESWL. METHODS: Between April 1996 and June 2003 9554 ESWL were performed at the lithiasis unit of the Fundacion Jimenez Diaz; 0.91% required previous insertion of a PN. We performed a retrospective analysis of the 49 patients with the diagnosis of upper urinary tract stones who require treatment by ESWL associated with PN. Indications for PN were: ureteral obstruction by fragments 6%, treatment of residual stones after percutaneous nephrolithectomy 6%, ureteral obstruction by a calcified double J stent 5%, urinary sepsis 75%, and obstructive anuria in a solitary kidney 8%. RESULTS: Overall, 87 sessions were performed in 49 patients. Number of shock waves 3996, Kv 7.69. Results where comparable in terms of stone size and composition. 57% of the patients were stone-free after one session, 24% after 2, and 19% required more than 2 sessions. There were two failures requiring surgery. CONCLUSIONS: Although in-situ ESWL is the treatment of choice for renoureteral lithiasis, the PN is a complementary procedure when ureteral obstruction requires treatment; it is non invasive and may be successfully associated to ESWL. PMID- 15536954 TI - [Treatment of a cavernous hemangioma of the bladder by endoscopical resection]. AB - OBJECTIVES: We report three cases of bladder hemangioma. METHODS: Three cases of cavernous hemangioma of the bladder with pathologic confirmation treated by endoscopical resection. RESULTS: There is no evidence of recurrence in either patient after resection. CONCLUSIONS: The endoscopical treatment of small bladder hemangiomas is an effective treatment. PMID- 15536955 TI - [The role of CT scan in the diagnosis of renal cell carcinoma]. AB - OBJECTIVES: To establish the role of CT scan as a reliable and effective method in the diagnosis of renal cell carcinoma (RCC) and to report the experience with this radiological test at the Manuel Fajardo Hospital. METHODS: 29 patients with the diagnosis of RCC who underwent CT scan during the period between January 1992 and December 2002 were evaluated. All patients were preoperatively classified as stages I to IV by CT scan. Tumor stage was confirmed by surgical findings and pathology report, and matched with preoperative CT scan evaluation in all cases. RESULTS: 29 patients with the diagnosis of RCC were treated, 65.5% were males and 34.5% females between 48 and 73 years of age. In 100% of the tumors were confirmed by ultrasound (US) and CT scan, and their limits were well defined. CT scan detected regional lymph nodes in 3.4% of the patients, distant lung metastases in two patients (6.8%) and bone metastases in one (3.4%). Preoperative study showed a tumor limited within the kidney in 72.3% of the patients, not exceeding Gerota's fascia in 17.2%, and extending outside Gerota's fascia in 10.3%. All patients underwent radical nephrectomy through an anterior approach and CT scan precision was confirmed, being quite similar to that of the surgical procedure. CONCLUSIONS: CT scan with or without contrast is the most precise method of diagnostic confirmation and staging currently available, achieving 100 percent of the diagnosis in comparison with other diagnostic modalities. PMID- 15536956 TI - [Spontaneous recanalization of the vas deferent after vasectomy: report of 2 new cases. Bibliographic review]. AB - OBJECTIVES: Vasectomy is a low morbidity and high efficacy surgical method. A small percentage of cases may have paternity after surgery as a complication. METHODS: We reviewed all patients who underwent vasectomy at our department from 1995 to 1999 (n = 1492), defining technical failure as pregnancy or no disappearance of spermatozoids in the follow-up sperm analysis. RESULTS: We found 2 cases; one of them did not have a negative sperm test after three months, the other one's wife became pregnant after previous azoospermia. Both patients showed granulomas in the pathologic report after a second operation. CONCLUSIONS: Vasectomy is a surgical technique that may have a failure rate close to 2%. Paternity after the operation may appear even after several years. Several authors report the existence of granulomas in those cases in which spontaneous recanalization appears. In our series it was a constant. PMID- 15536957 TI - [Multilocular cystic nephroma: diagnosis by imaging tests]. AB - OBJECTIVES: To describe the presenting signs of multilocular cystic nephroma (MCN) in various imaging tests. METHODS: We report a new case of MCN and describe the radiological characteristics of this neoplasia on intravenous urography (IVU), pyelography , CT scan and MRI. RESULTS: CT scan and MRI findings are highly suggestive of multilocular cystic nephroma. CONCLUSIONS: Various radiological tests may allow us to establish a working diagnosis for this kind of neoplasia, but CT scan and MRI are the main ones. PMID- 15536958 TI - [Penile hematoma secondary to rupture of the superficial dorsal vein of the penis]. AB - OBJECTIVES: To report a very unfrequent case of penile hematoma secondary to rupture of the superficial dorsal vein of the penis during intercourse. METHODS: 35-year-old patient presenting at the emergency department with a half moon shape hematoma in the dorsal part of the penis, not much painful, developed when initiating intercourse that we believe was a rupture of the superficial dorsal vein of the penis. RESULTS: In spite of the few cases of rupture of the superficial dorsal vein of the penis described, and after differential diagnosis with other causes of acute penis, conservative attitude with anti-inflammatory drugs and compressive bandage was undertaken. CONCLUSIONS: The rupture of the superficial dorsal vein of the penis is an unfrequent pathology which should be taken into differential diagnosis in a patient consulting with acute penile edema, the attitude is conservative with complete recovery without sequel. PMID- 15536959 TI - [Adrenal gland cyst. Report of one case and review of the literature]. AB - OBJECTIVES: To report one case of adrenal gland cyst in a patient presenting with lower urinary tract symptoms. The diagnosis and therapeutic aspects are discussed. METHODS: Diagnostic studies included ultrasound, KUB x-ray, CT scan and fine needle aspiration. RESULTS: Conservative treatment was chosen after the diagnosis of uncomplicated adrenal gland cyst with oligosymptomatic obstructive uropathy. CONCLUSIONS: The reliability of diagnostic tests (radiological and cytological) for adrenal cysts leave surgical treatment for symptomatic tumors and complicated cases; an incidental finding does not justify surgery. PMID- 15536960 TI - [Renal angiomyolipoma with arteriovenous fistula causing retroperitoneal hematoma. Case report]. AB - OBJECTIVES: To report the case of a female patient with ultrasound diagnosis of great left retroperitoneal hematoma admitted to the hospital for diagnostic workup and treatment. METHODS: 39-year-old patient presenting with intense epigastric pain irradiated to both flanks. Hematuria was not present. CT scan and arteriography led to the diagnosis of angiomyolipoma with arteriovenous fistula and subcapsular hematoma. Selective embolization was carried out. CONCLUSIONS: Hamartomas account for 3% of kidney tumors in adults; they are composed of 3 mesodermal tissues, including blood vessels. The lack of elastic membrane in their arteries favours aneurysm development. PMID- 15536961 TI - [Segmental renal infarction presenting as acute flank pain]. AB - OBJECTIVES: To report the clinical case of a male with acute onset right flank pain and significant deterioration of his general status. METHODS: 40-year-old male patient with acute onset of flank pain, progressive worsening, and fever up to 40 degrees C. Blood analysis (hemogram, coagulation tests, and serum ions), urine analysis, and abdominal x-rays, ultrasound and CT scan were performed. After the diagnosis of segmental renal infarction treatment with low molecular weight heparin was started. RESULTS: Patient improved clinically and was discharged from hospital 10 days later and received final discharge as outpatient after 6-month follow-up visit. CONCLUSIONS: Renal infarction should be taken into consideration in the differential diagnosis of acute flank pain, when more frequent causes as renal colic or inflammatory-infectious diseases are ruled out, mainly in patients with cardiovascular diseases because the most frequent cause is embolism. PMID- 15536962 TI - [Arterial priapism in a prepuberal patient. Conservative treatment]. AB - OBJECTIVES: To report the case of a 7-year-old patient with high flow priapism solved with conservative treatment. METHODS: We performed history and physical examination, complementary tests including general blood tests (serum ions, hemogram and coagulation tests), peripheral blood smear, Doppler ultrasound and selective arteriography which led to the diagnosis of high flow priapism. Due to the absence of arteriovenous fistula conservative treatment was chosen. RESULTS: Patient improved clinically after five days, with penile detumescence and spontaneous erections were preserved. CONCLUSIONS: Conservative treatment is a valid option in patients with arterial priapism, mainly in those cases in whom performing embolization of a small fistula seen on arteriography is not possible or when such AV communication is not demonstrated. PMID- 15536963 TI - Orthotopic bladder substitution in women with an ileocolonic pouch: functional and oncological outcome. AB - OBJECTIVES: In select female patients, bladder reconstruction following cystectomy will provide a better quality of life and self-image. We reviewed our results with ileocolonic orthotopic neobladders in female patients undergoing cystectomy for bladder cancer. Impact of pathologic stage on disease outcome, urinary continence results and surgical technique are described. Because of the relative paucity in the number of female neobladders performed, reporting of our results appear warranted. METHODS: Radical cystectomy with ileocolonic neobladder was performed in 22 consecutive women with bladder cancer over a five year period. Our technique spares the urethral support mechanism and innervation of the rhabdosphincter. Patients were selected based on renal function, ability to perform self-catheterization, organ confined disease and evidence of a disease free urethra, trigone and bladder neck. A retrospective review of the functional and cancer outcome was conducted. RESULTS: Postoperatively total diurnal urinary continence was achieved by 86% of the patients. Seventy-three percent of the patients had diurnal and nocturnal continence. Hypercontinence developed in 13.6% of patients. Only three patients developed advanced metastatic disease. None of the patients had evidence of local pelvic recurrence. The pathologic stage in the three patients that progressed were pT3a, pT3b, and pT2. CONCLUSIONS: Orthotopic neobladder substitution in female patients with bladder cancer is an alternative to a non-continent diversion. We report oncologic outcomes similar to the traditional anterior pelvic exenteration and non-continent urinary diversion. Surgical outcomes and complications are comparable with a significant improvement in the quality of life. PMID- 15536964 TI - A cry for help. PMID- 15536965 TI - Posting elder abuse alerts in physician offices. PMID- 15536966 TI - Physicians in politics: how hard can it be? PMID- 15536967 TI - The way things should go. PMID- 15536968 TI - Communication in the medical marriage. Part 2: Talking to your spouse. PMID- 15536969 TI - What every doctor should know about long-term care insurance, Part II. PMID- 15536970 TI - Health savings accounts--a revolutionary method of funding medical care expenses. AB - Health Savings Accounts coupled with a High-Deductible Health Care plan offer many eligible individuals the opportunity to reduce their insurance premiums and create a tax-deductible account to pay medical, dental, and vision care expenses. PMID- 15536971 TI - Health care decisions act--problems with consents, surrogacy, and end-of-life choices addressed by the new Tennessee Health Care Decisions Act. PMID- 15536972 TI - Monitoring quality of flexible endoscope disinfection by microbiologic surveillance cultures. AB - BACKGROUND: The purpose of this study is to assess the adequacy of currently approved high-level disinfection (HLD) for flexible endoscopes, by reviewing retrospectively the results of environmental cultures on the endoscopes, which were ready to use. METHODS: Cultures of all of our gastrointestinal endoscopes that were ready for use were sent to the laboratory for environmental cultures. Thirty milliliters of sterile water was passed through the biopsy channel of the endoscopes and the flushed water was collected in a sterile container. The specimen was plated onto blood agar, incubated at 37C, and examined for growth at 24 and 48 hours. RESULTS: Cultures were positive during the initial part of the study repeatedly with the same organism; the source of contamination was traced to be the source water and scope cleaner, after they were suspected and sent for cultures. In addition to Pseudomonas, there was contamination of the endoscopes with Micrococcus and Staphylococcus during the study period. CONCLUSIONS: Using environmental cultures for monitoring effectiveness of standard reprocessing procedures on a regular basis in clinical practice is not only inexpensive but also valuable to identify the source of possible contamination and rectify reprocessing methods to prevent transmission of infection secondary to endoscopy. PMID- 15536973 TI - Barriers to EMR adoption in internal medicine and pediatric outpatient practices. AB - BACKGROUND: Although electronic medical records (EMRs) are widely regarded as valuable tools in patient care, physicians in outpatient practices have been slow to adopt them. We sought to determine the current use of EMRs in area practices and identify physician attitudes related to their adoption. METHODS: Fax and mail survey of randomly selected physician representatives of all outpatient practices of Internal Medicine (n=51) and Pediatrics (n=24) in Shelby County, Tenn. Scores on eight physician attitudes regarding barriers to EMR adoption were obtained using a Likert scale. RESULTS: Survey response rate was 55%, with 18% reporting current EMR use. This corresponds to an EMR penetration of 20% for Shelby County. Current users were significantly less likely (P=0.005) than non-users to feel that an EMR interferes with doctor-patient interaction and less likely (P=0.019) to have EMR privacy concerns. While differences noted in other attitudes did not reach statistical significance, a trend was seen toward EMR users being less concerned (P=.0502) about reliability of an EMR. Large practices were no more likely than smaller ones to be using an EMR. Internal Medicine and Pediatric participants responded similarly to all items. The number of years in practice had no demonstrable impact on physician responses to these survey items. CONCLUSIONS: In this West Tennessee physician population, EMR user and non-user attitudes markedly differed about impact on doctor-patient interaction and patient privacy. If such concerns could be addressed to the satisfaction of physicians considering EMRs in their practice, adoption rates might be increased. PMID- 15536974 TI - Dangerousness in the home--violence against women. PMID- 15536975 TI - SAVE marks 10th anniversary. PMID- 15536977 TI - Compact multiphoton/single photon laser scanning microscope for spectral imaging and fluorescence lifetime imaging. AB - An inverted fluorescence microscope was upgraded into a compact three-dimensional laser scanning microscope (LSM) of 65 x 62 x 48 cm dimensions by means of a fast kHz galvoscanner unit, a piezodriven z-stage, and a picosecond (ps) 50 MHz laser diode at 405 nm. In addition, compact turn-key near infrared femtosecond lasers have been employed to perform multiphoton fluorescence and second harmonic generation (SHG) microscopy. To expand the features of the compact LSM, a time correlated single photon counting unit as well as a Sagnac interferometer have been added to realize fluorescence lifetime imaging (FLIM) and spectral imaging. Using this unique five-dimensional microscope, TauMap, single-photon excited (SPE), and two-photon excited (TPE) cellular fluorescence as well as intratissue autofluorescence of water plant leaves have been investigated with submicron spatial resolution, <270 ps temporal resolution, and 10 nm spectral resolution. PMID- 15536976 TI - Atomic force microscopy imaging of retroviruses: human immunodeficiency virus and murine leukemia virus. AB - Retroviruses are membrane-enveloped, RNA-containing viruses that produce a wide range of threatening diseases in higher animals. Among these are human immunodeficiency virus (HIV), which produces acquired immune deficiency syndrome (AIDS) in humans, and murine leukemia virus (MuLV), which produces leukemias in rodents. We have obtained the first atomic force microscopy (AFM) images of these two retroviruses, both isolated from culture media and emerging from infected cell surfaces. The HIV virions are 127 nm diameter on average, and those of MuLV are 145 nm, although there are wide distributions about the means. The AFM images show the arrangement of the envelope protein, responsible for host cell entry, on the surfaces of both virions. Disruption of the viruses using detergents or physical means allowed us to visualize interior structures, including the outer shells of both MuLV and HIV, the cores of MuLV, and the nucleic acid of HIV complexed with core proteins. Using immunolabeling techniques borrowed from electron microscopy, we were able to demonstrate the binding of gold-labeled antibodies directed against the envelope protein of MuLV. The AFM images are revealing, not only in terms of surface topology, but in terms of interior features as well, and they reveal the eccentricities and uniqueness of individual virus particles rather than yielding the average member of the population. Further application of AFM to viruses associated with other pathologies may ultimately have a significant impact on the diagnosis and treatment of virus promoted diseases. PMID- 15536978 TI - A novel technique for visualizing electron beam induced charging. AB - Charging is one of the most important problems encountered in scanning electron microscopy and as a result this phenomenon it has received a lot of both theoretical and experimental attention. Despite this, many questions remain about the nature and behavior of charging because of the limitations of the experimental techniques available to study it. For example, although it is now straightforward to determine in situ the surface potential of a sample that is charging during irradiation, it is difficult to measure the lateral extent of the charging, or its persistence once the incident beam is switched off. We describe here a simple technique which provides a rapid way of visualizing the temporal and spatial behavior of charging phenomena. PMID- 15536979 TI - Composition and stress analysis in Si structures using micro-raman spectroscopy. AB - Strained silicon (Si) technology enables improvements in complementary metal oxide semiconductor (CMOS) performance and functionality via replacement of the bulk, cubic-crystal Si substrate with an Si substrate that contains a tetragonally distorted, biaxially strained Si thin film at the surface. Here we use Raman spectroscopy to allow us to characterise growth processes of strained si, and to characterise the resulting level of strain/stress in the si and the effect it has on the underlying layer of graded SiGe. PMID- 15536980 TI - Secondary and backscattered electron imaging of weathered chromian spinel. AB - Secondary (SE) and backscattered electron (BSE) imaging as well as x-ray microanalysis have demonstrated that the weathering of chromian spinel occurs as a progressive form of alteration. Numerous chemical discriminant analysis methods based on the composition of chromian spinel are used to locate valuable deposits of minerals. These methods will be misleading if the correct interpretation of the weathering of chromian spinel and the subsequent pattern of changes in its mineral chemistry are not properly assessed using scanning electron microscopy. This assessment is vital in understanding the geological processes involved and the economic potential of any indicated deposit. Minerals such as chromian spinel, pyrope garnet, and picroilmenite are considered to be highly resistant to weathering and abrasion and are therefore useful in the search for associated valuable deposits of diamond, nickel, platinum, and gold. Known as indicator minerals, they are usually present in relatively large concentrations compared with the target mineral (e.g., diamond) and form large and often subtle dispersion anomalies adjacent to the deposit. Chromian spinel has long been regarded as a stable indicator mineral; however, detailed SE and BSE imaging indicates that many of the chromian spinels that are routinely examined using scanning electron microscopes (SEM) and microprobes are extensively altered. Secondary electron and BSE imaging of weathered chromian spinel in a normal SEM provides valuable data on the form and chemical style of the alteration. Secondary electron imaging of weathered chromian spinel in the environmental SEM (ESEM) not only enhances the difference in atomic number between unaltered and altered areas but also allows high-resolution imaging of these very fine replacement textures. PMID- 15536981 TI - Mechanical scanning of the specimen in the scanning electron microscope. AB - A scanning electron microscope (SEM) system equipped with a motor drive specimen stage fully controlled with a personal computer (PC) has been utilized for obtaining ultralow magnification SEM images. This modem motor drive stage works as a mechanical scanning device. To produce ultra-low magnification SEM images, we use a successful combination of the mechanical scanning, electronic scanning, and digital image processing techniques. This new method is extremely labor and time saving for ultra-low magnification and wide-area observation. The option of ultra-low magnification observation (while maintaining the original SEM functions and performance) is important during a scanning electron microscopy session. PMID- 15536982 TI - A simple method for checking the illumination profile in a laser scanning microscope and the dependence of resolution on this profile. AB - One of the conditions for a laser scanning microscope to reach its optimal performance is for it to operate at its full numerical aperture (NA). In most commonly used systems, the illumination intensity at the back focal plane of the objective lens is apodized. This paper presents a simple method using a photodiode for checking the actual illumination intensity profile. We show as an example the measured profiles of a laser beam when working with two high-NA immersion objectives in two different confocal systems, and also show that in theoretical studies of the point-spread function, the assumption of a flat compared with a truncated Gaussian beam profile gives rise to severe discrepancies. The measured profiles also serve as an indication of the necessity of a realignment of the optical system. PMID- 15536983 TI - Start up of deammonification process in one single SBR system. AB - A process for autotrophic nitrogen removal named aerobic/anoxic deammonification wherein NH4+ is oxidized by nearly 50% to NO2- and subsequently the ammonia is converted together with the nitrite to molecular nitrogen (N2 gas), has come to full-scale application within the last few years. In this research, sludge from a biological rotation disk located at a landfill leachate plant at Mechernich, Germany, which is capable of performing the deammonification process, was used as seed sludge for acclimating deammonification activities in laboratory scale batch reactors. In parallel, the same tests were performed with normal activated sludge. Research results indicated that deammonification activities could be obtained from the seeded reactor and also, with limited performance, from normal activated sludge in a single SBR system after several months acclimation. It was also seen that oxygen is an important factor that influences the deammonification from both the acclimatization process and process running. Further results were approved that report an impact of nitrite as a process intermediate on the closely related process of anaerobic ammonia oxidation ("Anammox"). However, limiting concentrations on a bacteria population performing deammonification were found to be different to those reported for a pure Anammox-culture. Also the influence of another intermediate, hydrazine, was tested for speeding up the acclimating process by inducing the deammonification activities and recovering the activities of deammonification from nitrite inhibition. PMID- 15536984 TI - Biological removal of NOx from flue gas. AB - BioDeNOx is a novel integrated physico-chemical and biological process for the removal of nitrogen oxides (NOx) from flue gas. Due to the high temperature of flue gas the process is performed at a temperature between 50-55 degrees C. Flue gas containing CO2, O2, SO2 and NOx, is purged through Fe(II)EDTA2- containing liquid. The Fe(II)EDTA2- complex effectively binds the NOx; the bound NOx is converted into N2 in a complex reaction sequence. In this paper an overview of the potential microbial reactions in the BioDeNOx process is discussed. It is evident that though the process looks simple, due to the large number of parallel potential reactions and serial microbial conversions, it is much more complex. There is a need for a detailed investigation in order to properly understand and optimise the process. PMID- 15536985 TI - Use of industrial wastewaters for the optimization and control of nitrogen removal processes. AB - In this experimental study the characterization of 2 industrial wastewaters, coming from an ice cream production industry (IW1) and a beet-sugar factory (IW2), with respect to their readily biodegradable fraction and denitrification potential, has been performed. To this end physical-chemical and biological characterization methods, both anoxic and aerobic, were used. Moreover a pilot scale SBR fed with municipal wastewater was started to verify the effect of the gradual addition of the concentrated organic wastewaters during the anoxic phase. The SBR was initially fed only with a primary municipal wastewater, then the organic load was increased by adding to the feed, during the anoxic phase, a small amount of the IW1 (second period). Once the initial conditions were restored the load was again raised using the second industrial wastewater (IW2) (third period). With those additions the nitrogen removal efficiency increased from 26% to 50%, in the case of the IW1 and from 23% to 53% in the case of the wastewater IW2, without any negative effect on the global performance of the system. In addition, periodical kinetic studies of denitrification and nitrification in the SBR, were performed. PMID- 15536986 TI - Nitrogen removal from pharmaceutical manufacturing wastewater via nitrite and the process optimization with on-line control. AB - In this study, laboratory scale experiments were conducted to investigate the nitrogen removal from pharmaceutical manufacturing wastewater. The results indicate that by selective inhibition of free ammonia on oxidizers, nitrogen removal can be achieved by nitritation and denitritation process. The nitrite ratio was above 98% in the aerobic stage and the nitrogen removal efficiency was about 99%. The complete ammonia removal corresponded exactly to the "Ammonia Valley" in the pH versus time graphic and the anoxic reaction was completed when the "Nitrite Knee" appeared in the ORP versus time graphic. Optimization of the SBR cycle by step-feed and on-line control with pH and ORP strategy allowed the carbon and energy saving. The easy operation and the low cost make the SBR system an interesting option for the biological nitrogen removal from the pharmaceutical manufacturing wastewater. PMID- 15536987 TI - Nitrogen removal from pharmaceutical manufacturing wastewater with high concentration of ammonia and free ammonia via partial nitrification and denitrification. AB - In this study, laboratory-scale experiments were conducted applying a Sequencing Batch Reactor (SBR) activated sludge process to a wastewater stream from a pharmaceutical factory. Nitrogen removal can be achieved via partial nitrification and denitrification and the efficiency was above 99% at 23 degrees C+/-1. The experimental results indicated that the nitrite oxidizers were more sensitive than ammonia oxidizers to the free ammonia in the wastewater. The average accumulation rate of nitrite was much higher than that of nitrate. During nitrogen removal via the nitrite pathway, the end of nitrification and denitrification can be exactly decided by monitoring the variation of pH. Consequently, on-line control for nitrogen removal from the pharmaceutical manufacturing wastewater can be achieved and the cost of operation can be reduced. PMID- 15536988 TI - Phosphorus removal under anoxic conditions in a continuous-flow A2N two-sludge process. AB - The Anaerobic-Anoxic/Nitrification (A2N) system is a continuous-flow, two-sludge process in which Poly-P bacteria are capable of taking up phosphate under anoxic conditions using nitrate as an electron acceptor. The process is very efficient because it maximizes the utilization of organic substrate for phosphorus and nitrogen removal. An experimental lab-scale A2N system fed with domestic sewage was tested over a period of 260 days. The purpose of the experiment was to examine phosphorus removal capacity of a modified A2N two-sludge system. Factors affecting phosphorus and nitrogen removal by the A2N system were investigated. These factors were the influent COD/TN ratio, Sludge Retention Time (SRT), Bypass Sludge Flow rate (BSF) and Return Sludge Flow rate (RSF). Results indicated that optimum conditions for phosphorus and nitrogen removal were the influent COD/TN ratio around 6.49, the SRT of 14 days, and the BSF and RSF were fixed at about 26 33% of influent flow rate. PMID- 15536989 TI - Nitrogen removal during leachate treatment: comparison of simple and sophisticated systems. AB - Membrane bioreactors (MBR) have become common in treating municipal wastewaters. Applied to leachates treatment MBR were also successful with pilot scale experiments and full-scale facilities as well. We succeeded previously in designing an efficient nitrification-denitrification process with an ethylene glycol byproduct as carbon source for denitrification. Moreover, an unexpectedly high inert COD removal efficiency was also observed in the full-scale MBR facility thereby making it possible to increase the operating time of the final GAC (Granulated Activated Carbon) adsorber. Since MBR are very sophisticated systems. Simpler and "lower" cost systems can also be considered. For example it is possible to nitrify leachates from sanitary landfill using a simple infiltration-percolation technique with a low energy cost. To validate previously published laboratory experiments, a semi industrial-scale pilot installation was installed at the Montzen landfill site (Belgium). The process is based on infiltration-percolation through a granular bed. This well known process was modified to increase the load, notably by changing the support medium, adding an electric fan that is run intermittently and maintaining temperatures greater than 15 degrees C. The new material is a type of granular calcium carbonate with a large specific surface area. These technical improvements enabled the system to nitrify up to 0.4 kg NH4+-N/m3 of reactor bed per day at a hydraulic load of 0.35 m.d(-1), with an ammonia removal rate in the range of 80 to 95%. Despite the high ammonia nitrogen inlet concentrations, this system exhibits remarkable nitrification efficiency. Moreover, these performances are achieved in a batch mode system without recirculation or dilution processes. If complete nitrification is needed, it can be obtained in a second in series of bioreactors. The system can be classified as a low cost process. An international patent is pending. Possible performances of those systems were compared with the usual methods for leachates treatment. PMID- 15536990 TI - Modeling response of nitrifying biofilm to inhibitory shock loads. AB - To study the response of nitrifying biofilm to inhibitory shock loads, a lab scale nitrifying biofilm reactor was operated in ambient conditions. Shock loads of various concentrations of inhibitory compound were applied to the biofilm. Aniline was used as an inhibitory compound. The experimental results were utilized to develop a model for predicting the variation of effluent nitrate concentration from the biofilm reactor for given shock loads of aniline concentration and exposure time both in exposure as well as in recovery phase. Close agreement between model and experimental observation of bulk aniline concentration and effluent nitrate concentration was obtained which indicates the usefulness of the model to estimate bulk aniline concentration and to predict the response of inhibitory shock loads on nitrifying biofilm. PMID- 15536991 TI - Freeze concentration for enrichment of nutrients in yellow water from no-mix toilets. AB - Separately collected urine ("yellow water") can be utilized as fertilizer. In order to decrease storage volumes and energy consumption for yellow water transport to fields, enrichment of nutrients in yellow water has to be considered. Laboratory-scale batch freeze concentration of yellow water has been tested in ice-front freezing apparatus: a stirred vessel and a falling film freeze concentrator (coolant temperatures: -6 to -16 degrees C). With progressing enrichment of the liquid concentrate, the frozen ice was increasingly contaminated with yellow water constituents (ammonia, total nitrogen, total phosphorus, TOC, and salts determined as conductivity). The higher the initial salinity of the yellow water and the lower the mechanical agitation of the liquid phase contacting the growing ice front, the more the frozen ice was contaminated. The results indicate, that in ice-front freezing devices multistage processes are necessary, i.e. the melted ice phase has to be purified (and the concentrates must be further enriched) in a second or even in a third stage. Energy consumption of this process is very high. However, technical scale suspension freeze concentration is reasonable in centralized ecological sanitation schemes if the population exceeds 0.5 million and distance of yellow water transportation to fields is more than 80 km. PMID- 15536992 TI - Biological nutrient removal model No.1 (BNRM1). AB - This paper presents the results of the work carried out by the CALAGUA Group on Mathematical Modelling of Biological Treatment Processes: the Biological Nutrient Removal Model No.1. This model is based on a new concept for dynamic simulation of wastewater treatment plants: a unique model can be used to design, simulate and optimize the whole plant, as it includes most of the biological and physico chemical processes taking place in all treatment operations. The physical processes included are: settling and clarification processes (flocculated settling, hindered settling and thickening), volatile fatty acids elutriation and gas-liquid transfer. The chemical interactions included comprise acid-base processes, where equilibrium conditions are assumed. The biological processes included are: organic matter, nitrogen and phosphorus removal; acidogenesis, acetogenesis and methanogenesis. Environmental conditions in each operation unit (aerobic, anoxic or anaerobic) will determine which bacterial groups can grow. Thus, only the model parameters related to bacterial groups able to grow in any of the operation units of a specific WWTP will require calibration. One of the most important advantages of this model is that no additional analysis with respect to ASM2d is required for wastewater characterization. Some applications of this model have also been briefly explained in this paper. PMID- 15536993 TI - Agricultural reuse of the secondary effluent polished by an algal pond system coupled with constructed wetland. AB - In this paper, reuse potentials of the secondary effluents as irrigation water was evaluated through field sampling trips. Water quality parameters significantly deviating from its guidelines were Total coliform bacteria and organic matter. Even though nutrients are not regulated in Korea, their removal would be required because they have been a barrier for secondary effluent irrigation through governmental intervention. The results of treatment study with aquatic ponds show that in spite of its poor biodegradability, organic matter in soluble form was reduced by 20% during 10 days of HRT. On the other hand, reduction of nutrients was remarkable. On average 85% of total nitrogen and 89% of total phosphorus were removed, respectively, which abates the worries of farmers for overgrowth or reduction in crop yield. However, coliform bacteria, although slightly reduced due to their propagation at the constructed wetlands still need further treatment. PMID- 15536994 TI - Calibration and simulation of two large wastewater treatment plants operated for nutrient removal. AB - Control and optimisation of plant processes has become a priority for WWTP managers. The calibration and verification of a mathematical model provides an important tool for the investigation of advanced control strategies that may assist in the design or optimization of WWTPs. This paper describes the calibration of the ASM2d model for two full scale biological nitrogen and phosphorus removal plants in order to characterize the biological process and to upgrade the plants' performance. Results from simulation showed a good correspondence with experimental data demonstrating that the model and the calibrated parameters were able to predict the behaviour of both WWTPs. Once the calibration and simulation process was finished, a study for each WWTP was done with the aim of improving its performance. Modifications focused on reactor configuration and operation strategies were proposed. PMID- 15536995 TI - Simultaneous organic stabilization and nitrogen removal in multi-stage biodrum system. AB - This paper presents the performance of a multi-stage biodrum system applied to domestic wastewater treatment. The organic stabilization and nitrogen removal efficiency in the system was investigated at different hydraulic retention times (HRT) of 12, 6 and 3 hours. The rotational speed of the biodrum was examined at 2,4 and 8 rpm. Average organic removal efficiencies in the system at different HRTs of 12, 6 and 3 hours were 96.3, 94.4 and 90.9%. Simultaneously, average nitrogen removal efficiencies were 91.5, 90.6 and 81.0%. The effect of rotational speed on nitrogen removal efficiencies in the system was clearly observed at a low HRT of 3 hours. The experimental results suggested that optimum HRT in the system was 6 hours. Moreover, they revealed that nitrogen removal efficiencies in the reactors operated at different rotational speed were in the same degree when considering the effluent nitrogen concentration. However, the reactors operated at lower rotational speed needed to employ higher numbers of biodrums (4 stages) than the others with higher rotational speed (3 and 2 stages at 4 and 8 rpm.) in order to achieve similar effluent qualities. At a rotational speed of 2 rpm, maximum nitrogen removal rate was found to be 0.2 kg/m3/d. PMID- 15536996 TI - Two-stage anaerobic digestion process for complete inactivation of enteric bacterial pathogens in human night soil. AB - Anaerobic digestion offers a good alternative for human waste treatment. However, the fate of enteric bacterial pathogens present in human night soil (HNS) remains a major concern for hygienic safety of the process. A two-stage anaerobic digestion process, consisting of separate acidogenic and methanogenic digesters, was designed and its efficacy in the inactivation of Salmonella typhi was compared to a single-stage digestion process. In a single-stage digestion, complete pathogen inactivation was achieved only in the digesters with high levels of volatile fatty acids (VFA approximately equal to 18,000 mg/l) and acidic pH (approximately equal to 6.0). These digesters, however, showed drastic reduction in methane yield. In the two-stage digestion process, S. typhi was completely inactivated in the acidogenic digester and the methanogenic digester was free from the pathogen even after receiving a daily dose of the pathogen. The process also achieved complete inactivation of other enteric pathogens, viz., Shigella dysenteriae and Vibrio cholerae. The two-stage process was efficient in biogas generation from HNS. Thus, the two-stage process ensures complete hygienic safety in anaerobic digestion of human night soil. PMID- 15536997 TI - Application of Methanobrevibacter acididurans in anaerobic digestion. AB - To operate anaerobic digesters successfully under acidic conditions, hydrogen utilizing methanogens which can grow efficiently at low pH and tolerate high volatile fatty acids (VFA) are desirable. An acid tolerant hydrogenotrophic methanogen viz. Methanobrevibacter acididurans isolated from slurry of an anaerobic digester running on alcohol distillery wastewater has been described earlier by this lab. This organism could grow optimally at pH 6.0. In the experiments reported herein, M. acididurans showed better methanogenesis under acidic conditions with high VFA, particularly acetate, than Methanobacterium bryantii, a common hydrogenotrophic inhabitant of anaerobic digesters. Addition of M. acididurans culture to digesting slurry of acidogenic as well as methanogenic digesters running on distillery wastewater showed increase in methane production and decrease in accumulation of volatile fatty acids. The results proved the feasibility of application of M. acididurans in anaerobic digesters. PMID- 15536998 TI - Separation efficiency in a whirlpool surface tension separator, separating faeces and toilet paper for nutrient recovery--pilot-scale study. AB - The main proportion of the plant nutrients in waste from society can be recycled in two unpolluted fractions if the urine and the faeces are collected separately. By using urine-diverting toilets combined with a whirlpool surface tension faecal separator, it is possible to achieve this. If the separator is installed correctly, with a gradual bend to minimise disintegration of the particles, it is possible to collect approximately 92% nitrogen, 86% phosphorus and 76% potassium of the content excreted in the faeces in a small separated fraction that only contains 10% of the flushwater used. The faecal separation is a robust system with no moving parts, which is not significantly affected by the flushwater volume, and almost no water is separated to the separated solids if neither toilet paper nor faeces are flushed. PMID- 15536999 TI - The large PAO cells in full-scale EBPR biomass samples are not yeast spores but possibly novel members of the beta-Proteobacteria. AB - Large, homogenous clusters of coccobacilli were found to be abundant in the biomasses from a conventional plant at Rosebud, Victoria, Australia. The identity and the in situ physiology of these dominant microorganisms were investigated in this study. These large clustered cells were revealed to be neither Gram positive nor Gram negative bacteria and contain polyP granules. Cells with similar features were also observed in some enhanced biological phosphate removal (EBPR) systems and reported as yeast spores and Rhodocyclus-related polyphosphate accumulating organisms (PAOs). In this study, fluorescent in situ hybridization (FISH) probing showed these cells were prokaryotic and members of the beta Proteobacteria. However, these large clustered cells did not respond to the PAO mix FISH probes. The in situ physiology of these large cells was studied with FISH in combination with microautoradiography (MAR) in order to understand their substrate assimilation abilities under different conditions as well as their phosphate uptake ability. These cells were able to take up acetate, glutamate and aspartate, but not glucose under both aerobic and anaerobic conditions. Nile Blue A staining in combination with MAR showed that cells incubated under anaerobic conditions contained polyhydroxyalkanoates (PHA) granules. In addition, MAR showed aerobic 33Pi assimilation with all these substrates, consistent with them supporting an EBPR capacity in these large cells. As well as raising doubts about a role for yeasts in EBPR, this study suggests that much still needs to be learned about the identity and level of biodiversity of the PAO in EBPR systems, and emphasizes the benefits of using techniques like FISH/MAR and PHA staining/MAR to resolve the in situ physiology of the populations of interest there. PMID- 15537000 TI - Optimising design, operation and energy consumption of biological aerated filters (BAF) for nitrogen removal of municipal wastewater. AB - The Biofiltration process in wastewater treatment combines filtration and biological processes in one reactor. In Europe it is meanwhile an accepted technology in advanced wastewater treatment, whenever space is scarce and a virtually suspended solids-free effluent is demanded. Although more than 500 plants are in operation world-wide there is still a lack of published operational experiences to help planners and operators to identify potentials for optimisation, e.g. energy consumption or the vulnerability against peakloads. Examples from pilot trials are given how the nitrification and denitrification can be optimised. Nitrification can be quickly increased by adjusting DO content of the water. Furthermore carrier materials like zeolites can store surplus ammonia during peak loads and release afterwards. Pre-denitrification in biofilters is normally limited by the amount of easily degradable organic substrate, resulting in relatively high requirements for external carbon. The combination of pre-DN, N and post-DN filters is much more advisable for most municipal wastewaters, because the recycle rate can be reduced and external carbon can be saved. Exemplarily it is shown for a full scale preanoxic DN/N/postanoxic-DN plant of 130,000 p.e. how 15% energy could be saved by optimising internal recycling and some control strategies. PMID- 15537001 TI - Treatment of landfill leachate by a pilot-scale modified Ludzack-Ettinger and sulfur-utilizing denitrification process. AB - Nitrogen removal efficiency of a pilot-scale system consisted of Modified Ludzack Ettinger (MLE) followed by sulfur-utilizing denitrification (SUDNR) process was evaluated with a landfill leachate. For SUDNR, a down-flow mode sulfur packed bed reactor (SPBR) filled with sulfur and limestone particles was used. Although total nitrogen removal efficiency of the MLE process was about 80% at the recycle ratio of 4, effluent contained 350-450 mg/L NO(3-)-N. Up to a loading rate of 1.2 kg NO(3-)-N/m3-day, the SPBR could achieve complete removal of nitrate, and nitrate removal rate was kept to that level even at higher loading rate. When a COD/N ratio of MLE process was maintained at 2 instead of 4, more organics with molecular weight less than 500 were utilized for heterotrophic denitrification although denitrification was not complete with the lack of electron donors. Clogging in the SPBR, mainly by the accumulation of nitrogen gas in the pores, could easily be removed by introducing the effluent in an upward direction for 1 min at 1 hr intervals. The proposed treatment system could achieve nitrate free effluent with a slight increase in chemical cost. Furthermore, depending on further COD removal requirement after biological treatment, the proposed treatment system can be an economical solution. PMID- 15537002 TI - Removal of ammonium from human urine through ion exchange with clinoptilolite and its recovery for further reuse. AB - Ammonium, from separately collected human urine, had been removed through transfer onto the ammonium selective natural zeolite, clinoptilolite, through ion exchange. In the subsequent treatment steps of washing with tap water, ammonium removed from urine was eluted from the surface of the clinoptilolite to be recovered for further reuse. Different quantities of clinoptilolite were used for a survey of the capacity of the zeolite for the process and to identify removal efficiencies based on initial ammonium loads. The highest surface concentration attained under experimental conditions employed was 15.44 mg ammonium per gram of clinoptilolite for an initial concentration of 110 mg ammonia per litre, and the highest removal was 98%, obtained for a loading of 1 mg ammonium per gram clinoptilolite. In the subsequent elution process, better removals were observed as pH was increased and the highest removal was attained at pH 13. The recovery was calculated as 9.73 mg ammonium per gram of clinoptilolite, corresponding to an efficiency of 63% only through washing with tap water. The results have given positive indications for the possibility of using ion exchange with clinoptilolite for the removal of ammonium from human urine and an incentive for improving methods of elution for its recovery for further reuse. PMID- 15537003 TI - Enhanced aerobic floc-like granulation and nitrogen removal in a sequencing batch reactor by selection of settling velocity. AB - In order to enhance the solid-liquid separation properties and nitrogen removal efficiency of SBR, the aerobic floc-like granules were cultivated under temporal alternating aerobic and anoxic conditions without the presence of carrier material in a SBR having 15 H/D (height/diameter) ratio. Two different effluent port positions were applied to the SBR for different selection of minimum settling velocities (over 0.6 and 0.7 m/h) of granules retained in the SBR during aerobic floc-like granule formation. The effect of different minimum settling velocities as an operational parameter on the size and solid-liquid separation properties of floc-like granules and also the COD and nitrogen removal of SBR were evaluated. The reactor was operated 6 hours per cycle (aerobic 4.75 hours, anoxic 1.25 hours) under chemical oxygen demand (COD) loading rate of 2.5 kg/m3 x d (1.3 kg acetate-COD and 1.2 kg glucose-COD). When increasing the minimum settling velocity by 0.1 m/h, the following results were observed at steady state. The nitrification efficiency was not changed at about 97% but the denitrification efficiency was improved from 78 to 97%. The COD removal efficiency was improved from 82 to 97% and the concentration of biomass in the reactor was retained at lower level at about 3,000 mg MLSS/L. The average sludge volume index (SVI) value of granules was decreased about 85 to 50 mL/g and the granule sizes were increased 0.1-0.5 mm to 1.0-2.0 mm. The required time to form granules and reach steady state was significantly shortened. Based on the results, the selection of the minimum settling velocity had a significant effect on both the physical properties of granules and the SBR performance, so it is suggested to use the minimum settling velocity as an operational parameter. PMID- 15537004 TI - Application of a sponge media (BioCube) process for upgrading and expansion of existing caprolactam wastewater treatment plant for nitrogen removal. AB - For the upgrade and expansion of an existing caprolactam wastewater treatment plant, a freely floating sponge media (BioCube) process was selected based on extensive pilot-plant tests, due to extreme space constraints. In order to protect nitrifier inhibition caused by high strength organics in caprolactam wastewater, the pilot plant consisted of an organics removal reactor, which functioned as a pretreatment for nitrification, and followed the nitrogen removal reactor. The suspended MLSS was 1,800-4,000 and the media attached MLSS was maintained at 22,000-26,000 mg/L. The final effluent COD was noticeably low, around 20.4-37 mg/L, even with fairly large fluctuations in the feed levels, between 1,400-6,770 mg/L. The removal of total nitrogen with the system, when denitrification was close to completion, was approximately 97.6%. For the entire run, complete nitrification of 99.6% was achieved, which might have been due to well-acclimatized nitrifiers attached in the BioCube media. Specifically, after adaptation, the nitrification continuously increased in the organics removal reactor, even under high residual organics conditions. From the numerous experimental results, the BioCube process seemed to be an effective method for the upgrading and expansion of the existing wastewater treatment plant, with minimum reactor enlargement. PMID- 15537005 TI - Control of nutrients after discharge to lakes through wastewater. AB - Control of nutrients as nitrogen and phosphorus after discharge into lakes is necessary since it is difficult and costly to control within wastewater plants currently in China. This paper studied the cycling of phosphorus and nitrogen with water and sediments from two lakes in China. It is found that oxygen plays a critical role in regulating phosphorus and nitrogen cycling within water and sediments. Three different oxygenation methods including aeration, calcium peroxide and hydrogen peroxide were studied to control phosphorus and nitrogen in overlying water. In anoxic conditions, the P concentration in water increased from an average 14 microg/L to 115.2 microg/L for Xili Lake, and from an average 24 microg/L to 1,000 microg/L for Jinchun Lake. The concentration of ammonia increased under anoxic conditions, while the concentration of nitrate increased under oxic conditions. In anoxic conditions, the nitrate concentration decreased probably through denitrification. Both N and P accumulation processes can be controlled under the three treatments. The phosphorus removal efficiency from the water body was in the order of CaO2 addition > aeration > H2O2 addition, while controlling effectiveness for ammonia was in the order of aeration > CaO2 addition > H2O2 addition. PMID- 15537006 TI - BICT biological process for nitrogen and phosphorus removal. AB - An updated biological nitrogen and phosphorus removal process--BICT (Bi-Cyclic Two-Phase) biological process--is proposed and investigated. It is aimed to provide a process configuration and operation mode that has facility and good potential for optimizing operation conditions, especially for enhancing the stability and reliability of the biological nutrient removal process. The proposed system consists of an attached-growth reactor for growing autotrophic nitrifying bacteria, a set of suspended-growth sequencing batch reactors for growing heterotrophic organisms, an anaerobic biological selector and a clarifier. In this paper, the fundamental concept and operation principles of BICT process are described, and the overall performances, major operation parameters and the factors influencing COD, nitrogen and phosphorus removal in the process are also discussed based on the results of extensive laboratory experiments. According to the experimental results with municipal sewage and synthetic wastewater, the process has strong and stable capability for COD removal. Under well controlled conditions, the removal rate of TN can reach over 80% and TP over 90% respectively, and the effluent concentrations of TN and TP can be controlled below 15 mg/L and 1.0 mg/L respectively for municipal wastewater. The improved phosphorus removal has been reached at short SRT, and the recycling flow rate of supernatant between the main reactors and attached growth reactor is one of the key factors controlling the effect of nitrogen removal. PMID- 15537007 TI - Nitrogen removal and sludge reduction in a symbiotic activated sludge system between anaerobic archaea and bacteria. AB - The possible symbiosis between bacteria and anaerobic archaea was investigated in intermittent aeration (I/A) systems. Archaea solution added to I/A reactor might play an important role in biological activities as well as in improvement of mineralization of organic matter. I/A reactor with archaea solution (I/A-arch) could increase both nitrification and denitrification rate and also reduce the sludge yield remarkably. These results indicate the possibility of the symbiotic activated sludge system with anaerobic archaea by controlling the DO level in the aeration tank. In this study, DO was controlled by intermittent aeration schemes and a successful symbiotic activated sludge system was achieved to reach the following conclusions. 1) SOUR of I/A-arch system was 2.9 mg-O2/g-VSS x min. SOUR and nitrification rate of the sludge from I/A-arch was higher than those from the I/A and A/S reactors. 2) Removal efficiencies of organic matter (TCOD(Cr)) in I/A arch, I/A and conventional activated sludge (A/S) reactors were 93, 90 and 87%, respectively. 3) Nitrification occurred successfully in each reactor, while denitrification rate was much higher in the I/A-arch reactor. Efficiencies of TN removal in A/I-arch, I/A and A/S reactors were 75, 63 and 33%, respectively. 4) Observed yield coefficients of I/A-arch, I/A and A/S reactors were 0.28, 0.41 and 0.37 g-VSS/g-COD. PMID- 15537008 TI - Automatic control and remote monitoring system for biological nutrient removal on small wastewater treatment plants in Korea. AB - Many small-size wastewater treatment plants in Korea's rural communities are designed to remove organic and suspended matter only, and they generally show a large fluctuation in the influent loading compared to municipal wastewater treatment plants (MWWTPs). They also have no professional engineers stationed for efficient operation against mechanical breakdown. For those reasons, the wastewater treatment plants have low efficiency in treatment of nitrogen and phosphorus as well as organic matter. In order to solve those problems, this study developed an automatic control system and RMS (remote monitoring system), which can keep efficiency stable despite any change in the small plants' loading rates and are capable of removing nutrient materials such as nitrogen or phosphorus. According to the results of the Experimental SBR system of the automatic control program, complete nitrification was made under oxic conditions and denitrification occurred as NO3-N concentration decreased by 0.5 mg/l in anoxic conditions and excellent nitrogen removal efficiency was seen generally. The Experimental SBR system created "phosphate release and uptake" effectively and displayed phosphate-removing efficiency up to more than 80% as the concentration of effluent was kept low by 0.4 mg/l. RMS developed in this study transmits a plant's data and operation states to clients in remote locations in real-time interval through the Internet. Therefore, although you are in a remote location, it allows you to see if a plant is properly operated or there is any breakdown. PMID- 15537009 TI - Nutrient removal and microbial granulation in an anaerobic process treating inorganic and organic nitrogenous wastewater. AB - The sustainable anaerobic nitrogen removal and microbial granulation were investigated by using a laboratory anaerobic granular sludge bed reactor, treating synthetic (inorganic and organic) wastewater and piggery waste. From inorganic synthetic wastewater, lithoautotrophic ammonium oxidation to nitrite/nitrate was observed by an addition of hydroxylamine. Also, the results revealed that the Anammox intermediates (particularly, hydrazine) contents in the substrate would be one of the important parameters for success of the anaerobic nitrogen removal process. The results from organic synthetic wastewater show that if the Anammox organism were not great enough in the startup of the process, denitritation and anaerobic ammonification would be a process prior to the Anammox reaction. The anaerobic ammonium removal from the piggery waste was performed successfully, probably due to the Anammox intermediates contained in the substrate. This reactor shows a complex performance including the Anammox reaction and HAP crystallization, as well as having partial denitritation occurring simultaneously. From the activity test, the maximum specific N conversion rate was 0.1 g NH4-N/g VSS/day (0.77 g T-N/g VSS/day), indicating that potential denitritation is quite high. The NO2-N/NH4-N ratio to Anammox is 1.17. The colour of the biomass treating the piggery waste changed from black to dark red. It was also observed that the red-colored granular sludge had a diameter of 1-2 mm. The settleability assessment of the granular sludge revealed that the granular sludge had a good settleability even though it was worse than that of seed granular sludge. PMID- 15537010 TI - Removal of nitrogen and phosphorus from industrial wastewaters by phytoremediation using water hyacinth (Eichhornia crassipes (Mart.) Solms). AB - This paper elucidates the phytoremediation potential of water hyacinth (Eichhornia crassipes [Mart.] Solms) for TN and TP rich industrial wastewaters determined for 15 weeks under different set-ups of 2-fold (56 TN mg/l and 15.4 TP mg/l), 1-fold, 1/2-fold, 1/4-fold and 1/8-fold and a control with no nutrients in duplicate. A mass balance was conducted to evaluate the phytoremediation efficiencies and to identify the key mechanisms of nutrient removal from the wastewaters. Our results manifested that water hyacinth is a promising candidate for a batch removal of TN and TP from wastewaters. 100% removal of both TN and TP was observed at the end of the 9th week in all the set-ups mainly due to assimilation and the period between 6-9weeks became the optimum period after which complete harvesting is recommended. Plants having an age of 6 weeks are ideal to commence the free-floating wetland and 21 days hydraulic retention time (HRT) is recommended for optimum removal of TN and TP. Assimilation and denitrification were the key mechanisms of TN removal while assimilation and sorption became the prominent mechanisms in the removal of TP from wastewaters. PMID- 15537011 TI - Simultaneous removal of nitrate and phosphate using cross-flow micellar-enhanced ultrafiltration (MEUF). AB - The feasibility of cross-flow micellar-enhanced ultrafiltration (MEUF) was investigated to remove nitrate and phosphate simultaneously. At the above critical micelle concentration (CMC), a cationic surfactant added in wastewater forms micelles, which have positive charge on their surface. Anionic contaminants such as nitrate and phosphate can be bound on the micelles by electrostatic interaction, and the micelle-pollutants complex is removed effectively by ultrafiltration. In this study, a cross-flow MEUF system was designed and investigated the feasibility of MEUF for field application. A cationic surfactant, cetylpyridinium chloride (CPC), was used, and the synthetic wastewater was treated by the polyacrylonitrile membranes with molecular weight cut-off (MWCO) of 30,000 Da and 10,000 Da. With the molar ratio of CPC to total pollutants of > 3, > 86% of nitrate and > 91% of phosphate were removed, respectively, and > 97% of CPC was also rejected. The flux was maintained 20-30% of the flux of distilled water. Therefore, it is feasible to remove nitrate and phosphate simultaneously using the cross-flow MEUF system. PMID- 15537012 TI - Enhanced granulation in UASB reactor treating low-strength wastewater by natural polymers. AB - Reetha (Sapindus trifoliata) seed extract and Chitosan were used as additives in the sludge bed of a UASB reactor treating low strength wastewater to enhance granulation. Five parallel laboratory scale UASB reactors were operated for 250 days with synthetic wastewater feed containing COD in the range of 600-800 mg/L. The reactors were seeded with spent sludge from a full-scale 5MLD UASB treatment plant at Jajmau, Kanpur, India. The seed sludge contained little or no granules. Different additives in the five reactors were as follows: control with no additive, cationic part of Reetha extract as additive, anionic part of Reetha extract as additive, bulk Reetha extract as additive and Chitosan as additive. The granulation rapidly increased in all the reactors beyond the 90th day of operation. The mean granule sizes as well as the fraction of granular sludge (particle size > or = 100 microm) were more in the presence of some of the additives compared to the control reactor. Chitosan significantly enhanced granulation followed by the cationic and anionic fractions of the Reetha extract. The bulk Reetha extract did not show enhancement of granulation. The ESEM/EDAX results showed that the bigger granules (3-4 mm) had porous structure and appeared as conglomerates of smaller granules. PMID- 15537013 TI - Modelling and calibration of phosphate and glycogen accumulating organism competition for acetate uptake in a sequencing batch reactor. AB - This paper improves the modeling of enhanced biological phosphate removal, by introducing the mechanistic description of a mixed culture of glycogen and phosphate accumulating organisms competing for acetate, and glycogen metabolism of the latter. A new process stoichiometry is defined for the mixed culture based upon previously developed metabolic concepts and models. The proposed model is tested with and used for the calibration of experimental data generated by a sequencing batch reactor operated with acetate feeding at a P/Ac ratio of 0.05 mgP mg(-1)COD. The calibration yielded acceptable numerical results for all the model coefficients tested and supported the validity of selected metabolic relationships for process stoichiometry. It also defined conditions that allow the system to sustain glycogen accumulating organisms with significant impact on process behaviour and performance. PMID- 15537014 TI - Model-based evaluation of nitrogen removal in a tannery wastewater treatment plant. AB - Computer modelling has been used in the last 15 years as a powerful tool for understanding the behaviour of activated sludge wastewater treatment systems. However, computer models are mainly applied for domestic wastewater treatment plants (WWTPs). Application of these types of models to industrial wastewater treatment plants requires a different model structure and an accurate estimation of the kinetics and stoichiometry of the model parameters, which may be different from the ones used for domestic wastewater. Most of these parameters are strongly dependent on the wastewater composition. In this study a modified version of the activated sludge model No. 1 (ASM 1) was used to describe a tannery WWTP. Several biological tests and complementary physical-chemical analyses were performed to characterise the wastewater and sludge composition in the context of activated sludge modelling. The proposed model was calibrated under steady-state conditions and validated under dynamic flow conditions. The model was successfully used to obtain insight into the existing plant performance, possible extension and options for process optimisation. The model illustrated the potential capacity of the plant to achieve full denitrification and to handle a higher hydraulic load. Moreover, the use of a mathematical model as an effective tool in decision making was demonstrated. PMID- 15537015 TI - The effect of anoxic selectors on sludge bulking. AB - The effect of the substrate feeding pattern on sludge settleability in denitrifying systems was investigated. The feeding pattern was simulated in two sequencing batch reactors (SBR). SBR1 simulates a system with a limiting substrate (acetate) and a non-limiting electron acceptor (nitrate), while SBR2 simulates the opposite condition. Sludge volume index (SVI) was the main observed parameter. The stoichiometry, substrate uptake rate, storage capacity as well as sludge morphology and mixed culture population dynamics were also determined. Results show that independent of the feeding pattern, the settleability of the sludge remained good (SVI approximately 50 mL/g) and stable. Neither the concentration of the acetate nor of the nitrate offered any limitations on sludge settleability, assuring that a good bio-P activity is present in nitrate limiting systems. Filamentous bacteria belonging to the TM group were usually present, although in low numbers. Despite the diffused, irregularly shaped and open floc structure, good settleability was observed. The results obtained were in line with the hypothesis on diffusion based selection. PMID- 15537016 TI - Treatment of nitrogen and phosphorus in highly concentrated effluent in SBR and SBBR processes. AB - Various sludge treatment processes produced supernatant with high ammonia concentration from 500 to 2,000 mgN/L and generally high phosphate concentration. Conversion of ammonia into nitrite via partial nitrification has proven to be an economic way, reducing oxygen and external COD requirements during the nitrification/denitrification process. Two processes with biomass retention are studied simultaneously: the sequencing batch reactor (SBR) and the sequencing batch biofilm reactor (SBBR). At a temperature of 30 degrees C, the inhibition of nitrite-oxidizing bacteria due to high ammonia concentration has been studied in order to obtain a stable nitrite accumulation. This work has confirmed the effect of pH and dissolved oxygen on nitrite accumulation performance. During a two month starting period, both processes led to nitrite accumulation without nitrate production when pH was maintained above 7.5. From a 500 mgN/L effluent, the performance of the SBR, and the SBBR, reached respectively about 0.95gN-NO2-/gN NH4+, and 0.4gN-NO2-/gN-NH4+. The SBBR appears to be more stable facing disturbances in dissolved oxygen conditions. Finally, the maximal phosphate removal rates obtained in the SBR reached 90%, and 70% in the SBBR, depending on ammonium accumulation in the reactor. Ammonium phosphate precipitation is likely to occur, as was suggested by crystals observation in the reactor. PMID- 15537017 TI - Monitoring toxicity of industrial wastewater and specific chemicals to a green alga, nitrifying bacteria and an aquatic bacterium. AB - Treatment plants may be exposed to a whole range of toxic organic and inorganic compounds that may inhibit the removal of organic matter and nitrogen. In order to secure maximum treatment efficiency, the plant manager has to monitor the toxicity of the influent sewage. With regard to the receiving water the manager also has to make sure that toxicity in the influent is significantly reduced during treatment. Because a whole range of chemicals may be present, chemical analysis may be insufficient and expensive as a control instrument. Instead, direct toxicity measurements are preferable to capture the complexity of the wastewater. The monitoring methods have to be relevant and sensitive for the processes in the treatment plant, i.e. removal of organic matter and nutrients. The methods also have to be simple and inexpensive. The paper reports on recent results from the application of nitrification, algae and Biotox tests, and summarises the experience with monitoring of toxicity. Although the sensitivity of the tests varies with respect to individual chemicals or group of chemicals, the application of a combination of the tests gives a high likelihood of detecting toxic impacts on treatment plants and receiving waters. PMID- 15537019 TI - Hybrid membrane systems for secondary effluent polishing for unrestricted reuse for agricultural irrigation. AB - Field experiments are in progress for secondary wastewater upgrading for unrestricted utilization for agricultural irrigation. The integrative approach of secondary effluent polishing is based on using a hybrid UltraFiltration (UF) and Reverse Osmosis (RO) membrane pilot system with a capacity of around 1 m3/hr. The UF effluent is used to feed the RO membranes. The RO permeate is subsequently applied for vegetable irrigation. Field results indicate the importance of the UF component in the removal of the organic matter and the pathogens that are still contained in the secondary effluent. Under specific conditions, when the dissolved solids content is relatively low, regarding sanitary and health aspects, the UF effluent can be applied for unrestricted irrigation. During the RO stage most nutrients are removed, allowing application of the effluent without jeopardizing the soil fertility and the aquifers. Preliminary economic assessment indicates that the extra cost for effluent polishing via the UF stage only is in the range of 5 to 15 US cents/m3. The extra cost for the RO stage is also assessed at 10 to 25 US cents/m3. The additional cost depends to a large extent on the quality of the incoming raw secondary effluent and local requirements of the command region. PMID- 15537018 TI - Experiences with a large-size WWTP based on activated sludge-biofiltration processes: 25 months of operation. AB - The South-Budapest Wastewater Treatment Plant (SBWWTP) had been operated as a high-load activated sludge (AS) plant since the middle of the 60s. According to the requirements proposed by the water authorities the treatment process had to be upgraded into nutrient (phosphorus and nitrogen) removal. The upgrade of the plant comprised implementation of BIOFOR type nitrifying (NP) and post denitrifying (DN) biofilters downstream of the AS stage. Phosphorus removal was obtained by chemical precipitation that can be done at five different points for feeding ferric-sulfate (Fe2(SO4)3). Partial flow recirculation was administered from the nitrifying BIOFOR unit ahead of the AS basin for pre-denitrification utilizing raw wastewater as carbon source. The plant performance was monitored since the test operation period for 25 months. Experience revealed that significant nitrification occurs in the high-load activated sludge basin originally designed for carbon removal. During the summer period (characterized by temperature of 20-25 degrees C) about 37-42% ammonium conversion rate was observed in the reactor. The decreasing temperature in the wintertime resulted in lower nitrification rates, of about 6-10%. The combined activated sludge biofiltration process proved its viability in the removal of organic matter, nitrogen and phosphorus. In this special configuration the AS system plays a key role in the nitrogen and organic matter removal. PMID- 15537020 TI - Nitrogen removal in a fluidized bed bioreactor by using mixed culture under oxygen-limited conditions. AB - Nitrogen removal involving nitrification and denitrification was investigated in a fluidized bed bioreactor by using mixed culture sludge under oxygen-limited conditions. Methane was used as a sole carbon source for denitrification. In this study, optimal nitrification and denitrification rates were examined by varying methane and oxygen gas dissolution flow rates, 90 ml/min, 400 ml/min and 650 ml/min, in each. Simultaneously nitrification and denitrification was achieved. The total nitrogen removal rate was 15-mg N/g VSS. d, 21-mg N/g VSS. d and 26.4 mg N/g VSS. d at gas dissolution flow rate 90 ml/min, 400 ml/min and 650 ml/min, respectively. No significant accumulation of nitrite was found in this experiment. Nitrogen removal rates depend on gas dissolution flow rates. DO concentration was at 0.5-2 mg/L. PMID- 15537021 TI - Nutrient removal from shrimp farm effluent by aquatic plants. AB - Duckweed was used to treat effluent from a shrimp farm in a batch wise and recirculation system. The result showed that duckweed could efficiently remove nutrients in the effluent, especially ammonia, which seemed to be the preferred nitrogen source of the plant. Red tilapia was used in the recirculation system and showed high potential in removal of uneaten shrimp feed. PMID- 15537022 TI - Organic and detergent degradation in combined O3/UF for domestic laundry wastewater reclamation. AB - This paper focuses on the evaluation of organic and detergent degradation in a combined Ozone/UF system for domestic laundry wastewater reclamation. Formation of by-product was investigated by GC/MS for the reclaimed water. Ozone was injected into the raw wastewater in a 10 L contact tank and the wastewater was circulated through the membrane module for inner pressurized cross-flow filtration. The concentrate was returned back to the contact tank. The membrane used in this experiment was hollow fiber polysulfone UF membrane with MWCO 10,000. It has an effective filtration area of 0.06 m2. The experiment was carried out with intermittent ozone injection, 5 min injection and 10 min idling. Ozone was dosed at the concentration of 1.5 mg/L. The flux of the UF could be maintained at 0.24 m/d under filtration pressure 40-45 kPa and water temperature, 20-22 degrees C. The organic removal efficiency by the system was 90% in terms of COD. Ozone was considerably effective to degrade organics in the wastewater. Molecular weight of organics in the raw waste was mostly greater than 10,000 (72% of 950 mgCOD/L). However 86% of effluent COD (94-100 mg/L) was composed of organics smaller than MWCO 500 by ozone injection. No harmful by-products by ozone contact were detected from the analysis of treated water using GC/MS. It was identified that residual organics in the treated water were 1,1' Oxybisbenzene, Octadecanoic acid, Squalene and Benzenmethanol, etc., which were additives contained originally in the detergent. Consequently the reclaimed water quality could be estimated safe enough to recycle for the rinsing cycle in a washing machine. PMID- 15537023 TI - Re-use of wastewater: preventing the recovery of pathogens by using medium pressure UV lamp technology. AB - Ultraviolet (UV) light has become widely accepted as an alternative to chlorination or ozonation for wastewater disinfection. There are now over 2,000 wastewater treatment plants worldwide using either low- or medium-pressure UV technology. Recent studies investigating UV lamp technology, configuration, cleaning requirements and ageing, as well as long-term performance tests, have demonstrated beyond any doubt the effectiveness of UV in inactivating pathogens in wastewater. Research has also shown that, to ensure permanent inactivation and prevent the recovery of microorganisms following exposure to UV, a broad, "polychromatic" spectrum of UV wavelengths is necessary. These wavelengths inflict irreparable damage not only on cellular DNA, but on other molecules, such as enzymes, as well. Only medium-pressure UV lamps produce the necessary broad range of wavelengths; low-pressure lamps emit a single wavelength peak which only affects DNA. Polychromatic medium-pressure UV light is so effective because of the lamp's exceptionally high UV energy output at specific wavelengths across the UV spectrum. It has been shown, for example, that pathogenic E. coli O175:H7 was able to repair the damage caused by low-pressure UV, but no repair was detected following exposure to UV from medium-pressure lamps. PMID- 15537024 TI - Energy saving processes for nitrogen removal in organic wastewater from food processing industries in Thailand. AB - Nitrogen removal from organic wastewater is becoming a demand in developed communities. The use of nitrite as intermediate in the treatment of wastewater has been largely ignored, but is actually a relevant energy saving process compared to conventional nitrification/denitrification using nitrate as intermediate. Full-scale results and pilot-scale results using this process are presented. The process needs some additional process considerations and process control to be utilized. Especially under tropical conditions the nitritation process will round easily, and it must be expected that many AS treatment plants in the food industry already produce NO2-N. This uncontrolled nitrogen conversion can be the main cause for sludge bulking problems. It is expected that sludge bulking problems in many cases can be solved just by changing the process control in order to run a more consequent nitritation. Theoretically this process will decrease the oxygen consumption for oxidation by 25% and the use of carbon source for the reduction will be decreased by 40% compared to the conventional process. PMID- 15537025 TI - The editorial, SADJ June 2004 and the CPD system. PMID- 15537026 TI - [Economic impact]. PMID- 15537027 TI - Scientific surface roughness values for resin based materials. AB - INTRODUCTION: Surface roughness of dental restorative materials is most often established with the Ra value obtained using profilometry or by assessing surface topography with the scanning electron microscope (SEM). Both methods should validate each other in confirming surface roughness. AIMS AND OBJECTIVES: The purpose of this study was to compare surface roughness values obtained with a profilometer to the SEM appearance of 6 resin-based restorative materials and assess whether Ra was appropriate as a sole surface roughness measure. Methods Six 5mm diameter specimen discs of Prodigy (Pr); Z100 (Z); Compoglass F (C); Hytac Aplitip (H); Photac-Fil (Pf) and Vitremer (V) were prepared against Mylar strips and stored in distilled water for 14 days. One side of each disc was sequentially polished with Soflex discs to super fine state, the other side remained unpolished. Three surface roughness measurements were made on each surface (n=18) recording Ra, Rv, Rp and Rt values, this data was subjected to a four way ANOVA and Tukey's Studentised Range Test (p=0.05). Two unpolished and two polished discs per material were prepared for SEM, evaluated and visually grouped for surface roughness. RESULTS: Approximate ascending order of roughness was Z, Pr, H, C, V, Pf for Ra, Rv, Rp and Rt and un/polished treatment. Polishing increases surimens into a "bland" (Pr, H, Z, C) and "textured" group (Pf and V). The polished specimens gave four groups: (Pr), (Z and C), (H) and (V and Pf) of increasing surface complexity. Polishing caused surface scratching, removed the matrix, reduced or removed filler particles and exposed voids within the material. CONCLUSIONS: This study emphasises the importance of using more than one technique to assess surface roughness. Rv and Rp values should be utilised to better understand polish induced surface feature changes. Rv maximum is a better measure to identify surface defects which could affect restoration longevity. PMID- 15537028 TI - Anti-bacterial efficacy of new self-etching dentine bonding agent. AB - Since a perfect seal is not easily achieved during bonding procedures, any product which will limit or delay bacterial proliferation in the resulting micro leakage spaces should extend the life span of bonded restorations. This study compared the antibacterial properties of an experimental, self-etching, fluoride releasing, antibacterial bonding system (ABF) with two standard bonding systems (SE Bond and Scotchbond MP). Spread plates of three different bacteria (Streptococcus mutans, Lactobacillus paracasei and Actinomyces naeslundii) were prepared on Brain-Heart Infusion agar. Standardised, sterilised filtration paper disks were infiltrated with 20 microl of primer and then placed on the inoculated agar and incubated at 37 degrees C for 48 hours. The extent of the inhibition zones were measured at different positions and data were analysed using the Student t-test to determine significant differences. All three primers showed zones of inhibition for all three bacteria tested. Inhibition zones for ABF primer against S. mutans were significantly larger (p<0.05) compared to that of Scotchbond MP and SE Bond. In general the antibacterial activity of the three primers against the three bacteria tested varied, and the inhibitory effect for the experimental primer was significantly superior against S. mutans only. PMID- 15537029 TI - Aspirin, other salicylates and non-steroidal anti-inflammatory agents. PMID- 15537030 TI - Alcohol: use, abuse and dependence. AB - As a drug, alcohol has grave bio-psychosocial effects on patients, their families and society in general. It is very important that clinicians be able to recognise, admit and thoroughly treat alcoholism. Subsequent to the indicated treatment of intoxication and withdrawal, the clinician should outline sound incentives (external initially, becoming internal in time) for rehabilitation and continued sobriety. A variety of alcohol-induced medical and psychiatric disorders should be recognised by doctors, and should be treated or referred appropriately. The multidisciplinary team should be involved, and doctors should not hesitate to seek help and advice. Alcoholism is potentially quite treatable. PMID- 15537031 TI - Paracetamol (acetaminophen) overdose. PMID- 15537032 TI - Oral health knowledge, attitudes and behaviour among nursing staff in Lesotho. AB - The decreasing prevalence of oral diseases in industrialised countries is attributed to a paradigm shift of oral health services from being mainly curative to preventative care. In under-resourced countries, the use of non-oral health care workers in the promotion of oral health, can contribute substantially to improving oral health and the adoption of a multidisciplinary team approach in oral health is highly recommended. Owing to the inadequate number of oral health personnel in Lesotho, the use of health care workers other than oral health care workers has been investigated. Nurses are one of the most suitable cadres of health care workers to take on this role. The aim of this study was to determine oral health knowledge, attitudes and behaviour among nurses in Lesotho. A descriptive cross-sectional study, using a random sampling method was used. A self-administered questionnaire with open and closed-ended questions was used to collect the data. The response rate was 87%. Knowledge of aetiology and prevention of dental caries, periodontal disease and the oral manifestations of HIV-infection was found to be adequate, while knowledge of oral cancer was found to be satisfactory. Nurses in Lesotho reported positive attitudes towards the provision of oral health education and oral hygiene practices. Most nurses themselves were found to be symptom-orientated in their utilisation of the oral health services. It is recommended that oral health be given greater attention in the nursing curriculum, with more clinical hands-on training in oral examination and diagnosis of oral diseases. PMID- 15537033 TI - Digital radiography: update for oral health care workers. AB - Digital Radiography is an imaging system that does away with the use of films. It constitutes an advance in computer technology and has made a significant impact on the field of Maxillofacial- and Dental Radiology. This paper presents an overview of the basic concepts and applications of dental digital radiography and compares it with conventional film-based imaging. In addition, it provides a thorough understanding of the direct, semi-direct and indirect dental digital imaging systems with their advantages and disadvantages. Universal acceptance of digital radiographic imaging as a diagnostic tool makes it important for oral health care workers to understand the principles thereof and to master the techniques involved in acquiring a diagnosable digital radiographic image. PMID- 15537034 TI - Essentials oils for implant maintenance: an evidence based appraisal. PMID- 15537035 TI - General practitioner's radiology case 24. Macrodontia of a single tooth. PMID- 15537038 TI - In praise of a pioneer: Franca Tantalo, RN, CNeph(C). PMID- 15537039 TI - Thirst distress and interdialytic weight gain: how do they relate? AB - Thirst is a frequent and stressful symptom experienced by hemodialysis patients. Several studies have noted a positive relationship between thirst and interdialytic weight gain (IDWG). These factors prompted us to consider ways that we could intervene to reduce thirst and IDWG through an educative, supportive nursing intervention. This paper presents the results of a pilot research project, the purpose of which was to: examine the relationship between thirst distress (the negative symptoms associated with thirst) and IDWG in a sample of our patients, describe patients' strategies for management of thirst, and establish the necessary sample size for the planned intervention study. The pilot research project results showed that in a small sample of 20, there was a mildly positive, though not statistically significant, correlation between thirst distress and IDWG (r = 0.117). Subjects shared a wide variety of thirst management strategies including: limiting salt intake, using ice chips, measuring daily allotment, performing mouth care, eating raw fruits and vegetables, sucking on hard candy and chewing gum. This pilot research project showed that given an alpha of 0.05 and a power of 80%, we will require a sample of 39 subjects to detect a 20% change in IDWG. We will employ these results to plan our intervention study, first by establishing the appropriate sample size and second by incorporating identified patient strategies into an educational pamphlet that will form the basis of our intervention. PMID- 15537040 TI - Diabetes mellitus Canadian clinical practice guidelines update. AB - Data from the National Diabetes Surveillance Strategy (NDSS) found the prevalence of physician-diagnosed diabetes in adults to be 4.8% (Health Canada Database, 2002). Population-based studies have found the prevalence rates to be much higher. The Canadian Diabetes Association has developed new clinical practice guidelines. In this article, I will attempt to highlight some of the major changes from the previously published guidelines. PMID- 15537041 TI - Evidence-based nursing practice. PMID- 15537042 TI - Research and the nephrology nurse. PMID- 15537043 TI - The nurse educator and research. PMID- 15537044 TI - Musings on nursing research. PMID- 15537045 TI - Profile of a new certified nephrology technologist: Meenakshi Sudarshan, CTech, CNephT. PMID- 15537046 TI - [Inequalities in health in Italy]. AB - Socioeconomic inequality and its impact on health is a growing concern in the European public health debate. In many countries, the issue is moving away from description towards the identification of the determinants of inequalities and the development of policies explicitly aimed at reducing inequalities in health. In Italy, ten years after the publication of the first report on inequalities in health, this topic is seldom present on the agenda of public policy makers. The purpose of this report is to update the Italian profile of social variation in health and health care in order to stimulate the debate on ways to tackle inequalities in health that are preventable. In the first section of this book, the threefold objective is to describe the principal mechanisms involved in the generation of social inequalities in health (Introduction); to report Italian data on the distribution and magnitude of this phenomenon in the last decade; and to evaluate policies and interventions in both the social (chapter 1.9, Section I) and the health sector (chapter 2.3, Section I), which are potentially useful to reduce health inequalities. It is intended for anyone who is in a position to contribute t o decision-making that will benefit the health of communities. For this reason, chapters are organized by specific determinants of inequalities on which interentions may have an impact. The methodological approach in the second section focuses on the best methods to monitor social inequalities including recommendations on social indicators, sources of information and study models, based on European guidelines revised for the Italian situation. According to data from national and local studies, mortality increases linearly with social disadvantage for a wide range of indicators at both the individual (education, social class, income, quality of housing) and the geographical level (deprivation indexes computed at different levels of aggregation). This positive correlation is evident for both sexes, with the steepest gradient observed among adults of working age, although differences persist also among the elderly. The causes of death found to be most highly correlated with social inequality, and largely responsible for the increasing inequality over the last decade, are those associated with addiction and exclusion (drug, alcohol and violence related deaths), with smoking (lung cancer) and with safety in the workplace and on the roads (accidents). Similar gradients and trends have been observed with different outcomes, such as self-reported morbidity, disability and cancer incidence (chapter 1.1, Section I). Reproductive outcomes confiirm this picture: compared to women belonging to the upper classes, those women in low conditions experience more spontaneous abortions and their children suffer from higher infant mortality and low birth weight. This is a critical issue since poor infant health, particularly for metabolic and respiratory pathologies, affects health in adult life. There is now substantive evidence showing that also socioeconomic circumstances at birth or during adolescence may have a strong impact on adult health (chapter 1.2, Section I). Differences in harmful lifestyles, such as smoking, heavy drinking, drug use, unhealthy diet, obesity and physical inactivity, have a similar effect. The only exception is smoking among women, which is positively correlated with socioeconomic status; however, since women in the upper classes have a greater tendency to quit smoking, the gradient will soon be reversed (chapter 1.7, Section I). On the other hand, most of these behaviours do not follow from free and conscious individual choice; they are a form of adaptation to chronic stress originating in the work-place (chapter 1.4, Section I), or to particularly unfavourable events and conditions, such as unemployment (chapter 1.5, Section I) or lack of family and social support (chapter 1.6, Section I). Poor socioeconomic circumstances are the threshold of absolute poverty and may lead to social exclusion, a condition with a heavy impact on health, which in Italy includes marginal groups of the native population and broader classes of immigrants (chapter 1.3, Section I). Finally, there is recent and consistent evidence on the existence of a "contextual" effect on health, as opposed to the "compositional" effect given solely by the aggregation of individual processes. According to this hypothesis, characteristics of the infrastructure, and the physical and socioeconomic environment of an area would have an impact on individual health independent from the cultural and economic resources personally available to people living in that area (chapter 1.8, Section I). With respect to the health care system, various studies are in agreement in demonstrating that poor and less educated people have inadequate access both to primary prevention and early diagnosis (chapter 2.1, Section I), and to early and appropriate care (chapter 2.2, Section I). They also experience higher rates of hospitalization, particularly in emergencies and with advanced levels of severity. PMID- 15537047 TI - Directed mobilization of endogenous neural progenitor cells: the intersection of stem cell biology and gene therapy. AB - Multipotential neural stem cells, which are capable of giving rise to both neurons and glia, line the cerebral ventricles of all adult animals, including humans. These cells may be mobilized and induced to undergo neuronal differentiation in vivo, by stimulating resident progenitor cells with both delivered and virally expressed growth factors. This strategy may be particularly efficacious in striatal neurodegenerative conditions such as Huntington's disease, in which lost medium spiny striatal neurons may be replenished through directed induction of progenitor cells lining the striatal ventricular wall. More broadly, our increasing understanding of the molecular control of progenitor cell mobilization and differentiation will likely afford many new opportunities for using induced neuronal replacement as a therapeutic strategy for neurodegenerative diseases. PMID- 15537048 TI - Safety of viral vectors for neurological gene therapies. AB - Viral-based gene therapy for neurological diseases provides a daunting challenge. Many neurological diseases, especially neurodegenerative conditions, have unknown etiologies and current viral platforms are largely inadequate for human therapy. The challenges appear surmountable through systematic definition and solution. This review highlights three problem areas for gene therapy; genomic integration, immunogenic responses and regulated gene expression, as well as current progress in those areas. Successful neurological gene therapy will require efficacy along with clear biosafety. Different vector platforms manifest characteristic properties that affect their suitability for human gene therapy and among these are their propensity to integrate, elicit immune responses and enable regulated gene expression. PMID- 15537049 TI - Clinical trials in neurological disorders using AAV vectors: promises and challenges. AB - Currently, there are five phase I clinical trials of recombinant adeno-associated viral vectors for the treatment of neurological disorders that are approved or likely to be approved shortly. Two trials are testing different strategies to treat Parkinson's disease (PD), the third trial is aimed at treating Canavan's disease, a pediatric leukodystrophy, the fourth trial targets Alzheimer's disease (AD), and the fifth will attempt to target the lysosomal storage disorder, Batten's disease. All four clinical trials rely on the de novo expression of an enzyme or a trophic factor to correct neuropathology. Ironically, the theories used to choose enzymes for the two PD trials were widely divergent, whereas the enzymatic strategy used for one of the PD trials and the Canavan's trial have remarkable similarities. Other gene therapy treatment strategies for PD and other disorders, such as amyotrophic lateral sclerosis, are also on the horizon. PMID- 15537050 TI - Viral vectors for gene transfer to striated muscle. AB - Recent progress has generated exciting results that are increasing the prospects for gene therapy of a variety of disorders of striated muscle, including the muscular dystrophies and myopathies, acquired and inherited diseases of cardiac muscle, and aging-associated muscle wasting. Numerous viral vector systems are being employed to transfer therapeutic genes to striated muscles, and advances in vector technology are leading to improved gene transfer efficiencies coupled with reduced immunological responses. The current technology in the field of viral vectors as tools for gene delivery to striated muscle is summarized, and recent developments related to gene therapies for skeletal and cardiac muscle disorders are discussed. PMID- 15537051 TI - Naked plasmid DNA for the treatment of muscular dystrophy. AB - Muscular dystrophy presents a formidable challenge for gene therapy. Major hurdles include the need to correct large masses of tissue (40% of the body weight) with minimal damage to the already inflamed and necrotic muscles, absence of immune rejection of the therapeutic protein, and sustained (if possible, life long) expression. Plasmid DNA has long been neglected as a candidate vector for this devastating disease, due to a low in vivo transfection efficiency. It, nevertheless, meets many of the prerequisites for a clinically viable treatment: ease of manufacturing, low toxicity, immunologically innocuous (to allow repeated administrations and insensitivity to pre-existing immunity), and accommodation of the large 11-kb dystrophin cDNA. Over the past year, interest has increased with two major breakthroughs: the first gene-based clinical trial for Duchenne muscular dystrophy that involved a human dystrophin plasmid, and a new method of intravascular delivery showing widespread transfection of limb muscles in large animals, including non-human primates. This opens a new avenue for the treatment of Duchenne dystrophy and many applications using gene delivery to skeletal muscle. PMID- 15537052 TI - Development of anthrax DNA vaccines. AB - Over 120 years ago, Pasteur and Greenfield developed an in vitro procedure for producing a live-attenuated Bacillus anthracis bacterial culture capable of protecting livestock from anthrax disease. Since then, anthrax has become one of the best characterized bacterial pathogens with regard to mechanism of toxicity and vaccine development. Most developments have used live-attenuated strains, bacterial supernatants or protein subunit approaches. Recently, novel plasmid DNA (pDNA) approaches to a safe and effective anthrax vaccine have been proposed. This review summarizes the history of anthrax, the need for new vaccines and recent developments in pDNA-based vaccines, leading to the initiation of a human phase I clinical trial in a significantly shorter timeframe than in traditional vaccine development. PMID- 15537053 TI - Gene therapy strategies to enhance the effectiveness of cancer radiotherapy. AB - Altering the genetic makeup of a cancer cell by gene transfer is a potentially powerful strategy for treating human cancer. However, a low efficiency of gene delivery in vivo and poor tumor specificity has prevented the widespread implementation of this technology in the clinic. Despite these formidable obstacles, the first successful application of gene therapy in the treatment of cancer could occur when it is combined with local modalities such as radiation therapy. A small number of gene therapy strategies have been evaluated in clinical trials in combination with external-beam radiation therapy. Combined therapy has been well tolerated and has not exacerbated the side effects of radiation therapy. Gene transfer and tumor cell destruction have been demonstrated in vivo. Although the results await confirmation in larger, prospective phase III trials, there is suggestive evidence that combined therapies may be demonstrating better than expected antitumor activity. The vast knowledge of the molecular defects that drive the cancer process, coupled with expanding the understanding of the genes responsible for tumor cell radioresistance, have led to the development of rational, targeted gene therapies designed to increase tumor cell radiosensitivity. Results of the clinical trials conducted to date are reviewed, followed by a description of new approaches currently under development. PMID- 15537054 TI - Gene therapy for autoimmune diseases. AB - Autoimmune diseases are threatening an increasing number of patients in developed countries, representing one of the major causes of disability and an enormous social cost. Current therapies mainly treat the symptoms of autoimmune diseases and are only partially able to interfere with disease evolution, and therefore decrease the degree of physical impairment. Thus, the development of new therapeutic strategies is imperative. This review focuses on gene therapy, as one possible alternative approach to the treatment of autoimmune disorders. The potential of gene therapy to specifically target tissues affected by autoimmune aggression, and its ability to interfere with the destructive pathogenic process while providing functional replacement and fostering reparative mechanisms will be emphasized. Gene therapy studies in experimental models of diabetes, rheumatoid arthritis and multiple sclerosis are reviewed. PMID- 15537055 TI - Inflammation-responsive promoters for fine-tuned gene therapy in rheumatoid arthritis. AB - The inflamed joints of rheumatoid arthritis (RA) patients are ideally suited for gene therapy applications that induce local production of potent anti inflammatory biologicals. The precise and absolute targeting needed when treating cancer is not necessary in RA. However, the challenge is to regulate transgene expression to meet variable physiological demands during the intermittent course of the disease in RA patients. Thus, a biosensing system with an inducible transcriptional switch that allows robust but adjustable transgene expression is required. Inflammation-inducible promoters are likely candidates to achieve precise control of transgene expression by physiologically driven processes. Acute-phase proteins, pro-inflammatory cytokines, heatshock proteins and hypoxia responsive genes are all related to the pathogenesis of RA, and their promoters can be exploited for disease-inducible transgene expression. With this, gene therapy enters a new era, that of temporal control of the therapeutic transgene expression. In addition to the reversible transcriptional switch, the ideal expression system also contains an amplification loop for high transgene expression and a drug-controllable switch to allow intervention by the physician. The merging of these modalities may provide a flexible system to fine-tune transgene expression, which is a prerequisite for the implementation of gene therapy in RA. PMID- 15537056 TI - Overview of target validation and the impact of oligonucleotides. AB - During target validation, researchers attempt to modulate the activity of potential drug targets in relevant cell and/or animal disease models in order to identify those that might be expected to have therapeutic benefit in human patients. This has become increasingly important with the large expansion of potential targets identified by the human genome project and as a result of the spiralling costs of bringing drugs to market. This review will present an overview of the target-validation mechanism and examine the strengths and weaknesses of using oligonucleotide-based technologies such as antisense, short interfering RNA and aptamers. PMID- 15537057 TI - Technology evaluation: AVI-4126, AVI BioPharma. AB - AVI BioPharma is developing AVI-4126, an antisense oligonucleotide targeted to c myc mRNA for the potential treatment of restenosis, cancer and polycystic kidney disease. AVI-4126 is currently undergoing phase II clinical trials. PMID- 15537058 TI - Technology evaluation: EMD-273063, EMD Lexigen. AB - EMD-273063 is an antibody/cytokine fusion protein comprised of a humanized version of the murine anti-GD2 antibody 14.18 coupled to two molecules of IL-2. It is being developed by EMD Lexigen Research Center Corp (a subsidiary of Merck KGaA) and its sister company, EMD Pharmaceuticals Inc, for the potential treatment of neuroblastoma and melanoma, for which it is in preclinical and phase I clinical trials, respectively. PMID- 15537059 TI - Technology evaluation: desmoteplase, PAION/Forest. AB - PAION GmbH, under license from Schering AG, is developing desmoteplase, a recombinant form of the anticlotting salivary plasminogen activator of the vampire bat Desmodus rotundus (DSPAalpha-1), for the potential treatment of acute ischemic stroke. The drug is currently undergoing phase II clinical trials. PMID- 15537060 TI - JCAHO and CMS to align quality measures: PI efforts will benefit. AB - Quality measures cover heart attack, heart failure, pneumonia, and surgical infection prevention. Using common metrics, data definitions will strengthen comparative data efforts. Some are concerned that tracking could affect decisions concerning sicker patients. PMID- 15537061 TI - MPEGs deliver message in patient safety program. AB - Video clips, each two to three minutes in length, address key goal or issue. All medical center staff were expected to embrace a culture of safety. Having commander endorse, introduce series proved an effective strategy. PMID- 15537062 TI - PASTE plus teams at core of award-winning program. AB - PASTE is a customized version of the Six Sigma DMAIC model. Best practices should not be taken at face value; you must have your own data to analyze. Develop a specific measurement plan for each project to accurately determine level of success. PMID- 15537063 TI - CA hospitals continue closing: a quality crisis? AB - The trend of hospital closings, now at eight, is expected to continue for the rest of the year. Uninsured patients, unfunded mandates are among key causes behind closings. Organization estimates more than half of all hospitals in California are operating in the red. PMID- 15537064 TI - WebM&M teaches by example with case studies. PMID- 15537065 TI - Shifts go up for bid: hospitals see boost in patient care, staff morale. AB - Having hospital staff fill all shifts leads to consistency of care, greater quality. Nurses can earn much more, while hospital outlays are lower without agency fees. Productivity also improves when in-house staff fill positions up for bid. PMID- 15537066 TI - Study: top performers also can be top improvers. AB - More than one-fourth of the variability on benchmark performance may be accounted for by sustained performance improvement. Hospitals were compared over several years using eight different performance measures. Most successful performers develop a culture of performance improvement. PMID- 15537067 TI - Study finds much higher rate of annual errors. AB - Institute of Medicine's use of health care worker's comments about charts may help explain discrepancies. Study indicates nearly dollars 3 billion in excess Medicare costs every year. Superior hospitals had five deaths per 1,000 less than the hospitals in the bottom 10th percentile. PMID- 15537068 TI - 'Most wired' hospitals widen gap over others. AB - More than 90% of most wired conduct return-on-investment analyses to justify expenditures. Computerized physician order entry is 10 times more likely to be used at most wired organizations than at least wired facilities. Information technology education of staff takes high priority at hospitals ranking high in survey. PMID- 15537069 TI - Tool locates alternative sites during bioterrorism. AB - Following a bioterror event, hospitals may be overwhelmed by an influx of patients. An Agency for Healthcare Research and Quality tool includes 30 different attributes by which to compare and rank facilities. The tool was made available to representatives at Olympics venues in Athens, Greece. PMID- 15537070 TI - Fast GC analysis with a 50 microm ID column: theory, practical aspects, and application to a highly complex sample. AB - This research focuses on the minimization of GC analysis times through the use of a 5 m x 0.05 mm ID x 0.05 microm (film thickness) column. Experimental minimum plate height (Hmin) and optimum linear velocity values were derived from standard compound applications, under various analytical conditions, and then related to classical chromatographic theory. Deviations from the latter are measured and discussed. Practical aspects linked to the use of such capillaries, such as column sample capacity and detector acquisition rates, are also considered. Furthermore, a fast, and what can be considered a very fast method, were applied to the separation of a fuel sample. Coefficients of variation of elution times and relative peak areas were calculated in the very fast application. All analytical results are compared with those obtained by conventional 0.25 mm ID column applications. PMID- 15537071 TI - Large volume splitless injection with concurrent solvent recondensation: keeping the sample in place in the hot vaporizing chamber. AB - An injector liner packed with a plug of glass wool is compared with a laminar and a mini laminar liner for large volume (20-50 microL) splitless injection with concurrent solvent recondensation (CSR-LV splitless injection). Videos from experiments with perylene solutions injected into imitation injectors show that glass wool perfectly arrested the sample liquid and kept it in place until the solvent had evaporated. The sample must be transferred from the needle to the glass wool as a band, avoiding 'thermospraying' by partial solvent evaporation inside the needle. The liquid contacted the liner wall when the band was directed towards it, but from there it was largely diverted to the glass wool. In the laminar liners, part of the liquid remained and evaporated at the entrance of the obstacle, while the other proceeded to the center cavity. Vapors formed in the center cavity drove liquid from the entrance of the obstacle upwards, but the importance of such problems could not be verified in the real injector. Some liquid split into small droplets broke through the obstacle and entered the column. Breakthrough through the laminar liners was confirmed by a chromatographic experiment. An improved design of a laminar liner for large volume injection is discussed as a promising alternative if glass wool causes problems originating from insufficient inertness. PMID- 15537072 TI - Separation, pre-concentration, and HPLC analysis of methylxanthines in urine samples. AB - An SPE method, using RP18 phases, for the simultaneous extraction of caffeine, theobromine, theophylline, paraxanthine, 1-methylxanthine, 3-methylxanthine, 7 methylxanthine, 1-methyluric acid, 1,3-dimethyluric acid, 1,7-dimethyluric acid, and 1,3,7-trimethyluric acid from urine has been developed. Besides a gradient HPLC system for the analysis of the compounds of interest on a LiChrosorb RP-18 (7 microm) column with mobile phase containing 0.05% aq. solution of trifluoroacetic acid and acetonitrile has been elaborated. The procedure has been successfully applied to the analysis of methylxanthines and methyluric acids in urine of patients with chronic asthma treated with theophylline and in urine of healthy subjects. PMID- 15537073 TI - Coupled-column liquid chromatography method with photochemically induced derivatization for the direct determination of benzoylureas in vegetables. AB - The coupled-column (LC-LC) system, consisting of a first column packed with internal surface reversed phase (ISRP) (50 x 4.6mm ID) and a Chrompack C18 (100 x 4.6 mm ID) as second column, allowed the simultaneous determination of five benzoylurea insecticides in dichloromethane (CH2Cl2) extracts of vegetable samples without any clean-up step. This system was combined with a photochemically induced fluorescence (PIF) post-column derivatization in order to provide strongly fluorescent photoproducts from the non-fluorescent benzoylureas. Limits of detection ranged from 0.21 to 0.98 microg L(-1) of pesticide (equivalent to 0.14-0.65 microg kg(-1) in vegetable samples) and limits of determination ranged from 4.0 to 10.0 microg L(-1) (equivalent to 2.7-6.7 microg kg(-1)). Linearity of the method was established between 2 and 1800 microg L(-1), depending upon the compound. Validation of the total method was performed by randomly analyzing recoveries of four vegetable samples (aubergine, cucumber, green bean, and tomato) spiked at two levels of concentration (10.0 and 33.3 microg kg(-1)). The combination of the LC-LC system with PIF detection provides a sensitive, selective, and rapid method for the determination of pesticides in vegetable samples at levels lower than the maximum residue levels (MRLs) established for these compounds by Spanish legislation. PMID- 15537074 TI - Development of a validated HPLC method for the determination of B-complex vitamins in pharmaceuticals and biological fluids after solid phase extraction. AB - An HPLC method was developed for the simultaneous determination of seven water soluble vitamins, viz. thiamine, riboflavin, nicotinic acid, nicotinamide, pyridoxine, cyanocobalamin, and folic acid, in multivitamin pharmaceutical formulations and biological fluids (blood serum and urine). Separation was achieved at ambient temperature on a Phenomenex Luna C18 (150 x 4.6 mm) analytical column. Gradient elution was performed starting at a 99:1 A:B v/v composition, where A: 0.05 M CH3COONH4/CH3OH (99/1) and B: H2O/CH3OH (50/50), at a flow rate of 0.8 mL/min. After a 4-min isocratic elution the composition was changed to 100% of B in 18 min and elution continued isocratically for 8 min. Detection was performed with a photodiode array detector at 280 nm. Each vitamin was quantitatively determined at its maximum wavelength. Spectral comparison was used for peak identification in real samples. Detection limits were in the range of 1.6-3.4 ng, per 20-microL injection, while linearity held up to 25 ng/microL. Accuracy, intra-day repeatability (n = 6), and inter-day precision (n = 7) were found to be satisfactory. Theobromine (2 ng/microL) was used as internal standard. Sample preparation of biological fluids was performed by SPE on Supelclean LC-18 cartridges with methanol-water 85/15 v/v as eluent. Extraction recoveries from biological matrices ranged from 84.6% to 103.0%. PMID- 15537075 TI - Optimization of a high-performance liquid chromatography system by artificial neural networks for separation and determination of antioxidants. AB - A high-performance liquid chromatography (HPLC) system was used to determine the antioxidants tert-butyl-hydroquinone (TBHQ), tert-butylhydroxyanisole (BHA), and 3,5-di-tert-butylhydroxytoluene (BHT) simultaneously in oils. The paper presents a new methodology for the optimized separation of antioxidants in oils based on the coupling of experimental design and artificial neural networks. The orthogonal design and the artificial neural networks with extended delta-bar delta (EDBD) learning algorithm were employed to design the experiments and optimize the variables. The response function (Rf) used was a weighted linear combination of two variables related to separation efficiency and retention time, according to which the optimized conditions were obtained. The above-mentioned antioxidants in rapeseed oils were separated and determined simultaneously under optimized conditions by HPLC with UV detection at 280 nm. Linearity was obtained over the range of 10-200 microg/mL with recoveries of 98.3% (TBHQ), 98.1% (BHT), and 96.2% (BHA). PMID- 15537076 TI - Enantiomer separation of dimethyl dicarboxy alpha-biphenyl (DDB) and its analogues on a covalently bonded cellulose tris-(3,5-dimethylphenylcarbamate) CSP. AB - Dimethyl dicarboxy alpha-biphenyl (DDB) and its analogues represent atropisomers which have been resolved on the covalently bonded cellulose tris-(3,5 dimethylphenylcarbamate) (CDMPC) CSP. Different kinds of alcohols, tetrahydrofuran (THF), and chloroform were employed as mobile phase modifiers (MPMs), and their influence on the retention and separation of the enantiomers was investigated. Ternary mobile phases (hexane/2-propanol/THF, hexane/2 propanol/chloroform) were employed to investigate the separation of the five enantiomers. The advantages of the broader choice of solvents offered by the covalently bonded CDMPC CSP were discussed. The effect of structural variation of the enantiomers on their retention and separation was investigated. PMID- 15537077 TI - On-line separation for the speciation of mercury in natural waters by flow injection-cold vapour-atomic absorption spectrometry. AB - Inorganic mercury and methylmercury are determined in natural waters by injecting the filtered samples onto a low cost commercial flow injection system in which an anion exchange microcolumn is inserted after the injection loop (FIA-IE). If hydrochloric acid is used as the carrier solution, the HgCl4(2-) species (inorganic mercury) will be retained by the anion exchanger while the CH3HgCI species (methylmercury) will flow through the resin with negligible retention. Four anion exchangers and seven elution agents were checked, in a batch mode, to search for the best conditions for optimal separation and elution of both species. Dowex M-41 and L-cysteine were finally selected. Mercury detection was performed by cold vapour-electrothermal atomic adsorption spectrometry (HG ETAAS). Both systems were coupled to perform the continuous on-line separation/detection of both inorganic mercury and methylmercury species. Separation and detection conditions were optimized by two chemometric approaches: full factorial design and central composite design. A limit of detection of 0.4 microg L(-1) was obtained for both mercury species (RSD < 3.0% for 20 microg L( 1) inorganic and methylmercury solutions). The method was applied to mercury speciation in natural waters of the Nerbioi-lbaizabal estuary (Bilbao, North of Spain) and recoveries of more than 95% were obtained. PMID- 15537078 TI - Micellar electrokinetic chromatography with laser-induced fluorescence detection for sensitive determination of ephedrine and pseudoephedrine. AB - A selective and sensitive micellar electrokinetic chromatography method with laser-induced fluorescence detection was developed for the quantification of ephedrine (E) and pseudoephedrine (PE) derivatized with 4-chloro-7-nitrobenzo-2 oxa-1,3-diazole. After conducting a series of optimizations, a running buffer of 10 mM sodium borate + 16 mM SDS was used for separation of the derivatives. A linear relationship for E and PE was obtained in the range of 0.044-6.6 microg mL(-1) (correlation coefficient: 0.9943 for E, 0.9946 for PE), and the detection limits for E and PE were 0.70 and 0.30 ng mL(-1), respectively. The sensitivity of E and PE was improved by several multiples of ten over those of CZE-LIF method. The method was applied to the analysis of the two alkaloids in ephedra herbal medicine and preparations with recoveries in the range of 98.3-107.1%. PMID- 15537079 TI - Immobilized betulinic acid column and its interactions with phospholipase A2 and snake venom proteins. AB - Betulinic acid (BA) is a plant-derived pentacyclic triterpenoid. Although BA has been found to have diverse pharmacological effects, including anti-tumor and anti inflammatory actions and potential as inhibitor of phospholipase A2 (PLA2), its cellular targets remain unclear. In this study, BA was immobilized onto an acrylamide matrix. The immobilized-BA column could retain the purified PLA2 of bovine pancreas or the PLA2 of snake venom from Naja nigricollis. The bound PLA2 were not eluted by high salt concentrations but were eluted by either acid or calcium free buffer. Besides the PLA2, a group of basic proteins of snake venom with molecular weights of about 7 kDa were also strongly bound by immobilized BA. One of these proteins was identified as gamma-cardiotoxin. The usefulness of immobilized BA for exploring the cellular targets of BA is discussed. PMID- 15537080 TI - Liquid chromatography analysis of monosubstituted sulfobutyl ether-beta cyclodextrin isomers on porous graphitic carbon. AB - The retention behaviour of the three positional isomers of monosubstituted sulfobutyl ether-beta-cyclodextrin was investigated on a porous graphitic carbon (PGC) column. The influence of the mobile phase composition (nature and concentration of organic and electronic modifiers) was studied as well as the effect of column temperature. These hydrophilic and anionic analytes were highly retained on the PGC stationary phase compared to octadecyl bonded phases. The retention is mainly governed by a reversed-phase mechanism with electronic interaction playing a secondary role. An increase in solute retention and efficiency with temperature was observed. Successful isocratic separation with satisfactory baseline resolution of the three isomers of monosubstituted sulfobutyl ether-beta-cyclodextrin was achieved at 75 degrees C on a Hypercarb column by using ammonium acetate as electronic modifier in water-acetonitrile (83:17). The chromatographic methodology developed can be easily used for relative quantification of each isomer within a mixture and can be applied for semi-preparative purification of each one. The evaporative light scattering detector allows the detection of these non UV-visible absorbing molecules. PMID- 15537081 TI - Packing capilllary electrochromatography columns using vacuum--a preliminary study. AB - This paper introducesa novel method for packing Capillary Electrochromatography Columns (CEC). Using vacuum packing methodology, silica particles as small as 1 microm were successfully packed into the capillary columns with 75 microm inner diameter. The columns are verystable and show no noticeable loss in efficiency after 200 sample injections. The performance of these vacuum packed capillary columns was evaluated with a mixture of aromatic and non-aromatic compounds. A 24 cm long capillary column can produce peak efficiencies of around 45,000 plates for benzene. PMID- 15537082 TI - Introductory remarks: bacterial endosymbionts or pathogens of free-living amebae1. AB - Free-living amebae are ubiquitous in the environment and can be isolated from a variety of habitats including water, soil, air, hospital water systems, dental units, contact lens cases, and cooling towers. The interaction of amebae with other microorganisms in their environment is varied. Bacteria are a major food source for free-living amebae. However, some bacteria have established a stable symbiotic relationship with amebae. Recent reports indicate an association of amebae with intracellular bacterial pathogens. Such amebae may serve as reservoirs for maintaining and dispersing pathogenic bacteria in the environment or as vectors of bacterial disease in humans. PMID- 15537083 TI - Genetic and physiological interactions in the amoeba-bacteria symbiosis. AB - Amoebae of the xD strain of Amoeba proteus that arose from the D strain by spontaneous infection of Legionella-like X-bacteria are now dependent on their symbionts for survival. Each xD amoeba contains about 42,000 symbionts within symbiosomes, and established xD amoebae die if their symbionts are removed. Thus, harmful infective bacteria changed into necessary cell components. As a result of harboring X-bacteria. xD amoebae exhibit various physiological and genetic characteristics that are different from those of symbiont-free D amoebae. One of the recent findings is that bacterial symbionts control the expression of a host's house-keeping gene. Thus, the expression of the normal amoeba sams gene (sams1) encoding one form of S-adenosylmethionine synthetase is switched to that of sams2 by endosymbiotic X-bacteria. Possible mechanisms for the switching of sams genes brought about by endosymbionts and its significance are discussed. PMID- 15537084 TI - Bacterial endosymbionts of free-living amoebae. AB - The occurrence of bacterial endosymbionts in free-living amoebae has been known for decades, but their obligate intracellular lifestyle hampered their identification. Application of the full cycle rRNA approach, including 16S rRNA gene sequencing and fluorescence in-situ hybridization with 16S rRNA-targeted oligonucleotide probes, assigned the symbionts of Acanthamoeba spp. and Hartmannella sp. to five different evolutionary lineages within the Proteobacteria, the Bacteroidetes, and the Chlamydiae, respectively. Some of these bacterial symbionts are most closely related to bacterial pathogens of humans, and it has been suggested that they should be considered potential emerging pathogens. Complete genome sequence analysis of a chlamydia-related symbiont of Acanthamoeba sp. showed that this endosymbiont uses similar mechanisms for interaction with its eukaryotic host cell as do the well-known bacterial pathogens of humans. Furthermore, phylogenetic analysis suggested that these mechanisms have been evolved by the ancestor of these amoeba symbionts in interplay with ancient unicellular eukaryotes. PMID- 15537085 TI - Ancient weapons for attack and defense: the pore-forming polypeptides of pathogenic enteric and free-living amoeboid protozoa. AB - Pore-forming polypeptides have been purified from several amoeboid protozoans that are well-known human pathogens. Obligate enteric parasites, such as Entamoeba histolytica, and free-living but potentially highly pathogenic species, such as Naegleria fowleri, contain these cytolytic molecules inside cytoplasmic granules. Comprehensive functional and structural studies have been conducted that include isolation of the proteins from their natural sources, monitoring of their biological activity towards different targets, and molecular cloning of the genes of their precursors. In the case of the most prominent member of the protein family, with respect to protozoans, the three-dimensional structure of amoebapore A was solved recently. The amoebic pore-forming polypeptides can rapidly perforate human cells. The antibacterial activity of amoebapores and of related polypetides from free-living protozoa points to a more vital function of these molecules: inside the digestive vacuoles they combat growth of phagocytosed bacteria which are killed when their cytoplasmic membranes are permeabilized. The concommitant activity of these proteins towards host cells may be due to a coincidental selection for an efficient effector molecule. Nonetheless, several lines of evidence indicate that these factors are involved in pathogenesis of fatal diseases induced by amoeboid protozoa. PMID- 15537086 TI - Modulation of a "CD59-like" protein in Naegleria fowleri amebae by bacteria. AB - Found in soil and freshwater habitats, Naegleria fowleri are free-living amebae that cause a fatal disease in humans called Primary Amebic Meningoencephalitis. In the natural environment, amebae feed on bacteria. In the infected host, the amebae lyse and ingest nerve tissue. Recently, we have established that N. fowleri expresses a "CD59-like" surface protein, but the function of this protein in the ameba has not been elucidated. In mammalian cells, CD59 is a complement regulatory protein that inhibits complement-mediated lysis of cells expressing this protein. In the present study, expression of the "CD59-like" protein in response to bacteria and bacterial toxins was investigated by Western immunoblot analysis. Co-culture of N. fowleri with log phase Escherichia coli or Pseudomonas aeruginosa resulted in differential expression of the "CD59-like" protein. Co cultures of amebae and bacteria were examined by electron microscopy. The results of our study implicate a possible protective role of the "CD59-like" protein in response to bacterial predators and bacterial toxins, because amebae remained intact after co-culture with bacteria. PMID- 15537087 TI - Plastid-targeting peptides from the chlorarachniophyte Bigelowiella natans. AB - Chlorarachniophytes are marine amoeboflagellate protists that have acquired their plastid (chloroplast) through secondary endosymbiosis with a green alga. Like other algae, most of the proteins necessary for plastid function are encoded in the nuclear genome of the secondary host. These proteins are targeted to the organelle using a bipartite leader sequence consisting of a signal peptide (allowing entry in to the endomembrane system) and a chloroplast transit peptide (for transport across the chloroplast envelope membranes). We have examined the leader sequences from 45 full-length predicted plastid-targeted proteins from the chlorarachniophyte Bigelowiella natans with the goal of understanding important features of these sequences and possible conserved motifs. The chemical characteristics of these sequences were compared with a set of 10 B. natans endomembrane-targeted proteins and 38 cytosolic or nuclear proteins, which show that the signal peptides are similar to those of most other eukaryotes, while the transit peptides differ from those of other algae in some characteristics. Consistent with this, the leader sequence from one B. natans protein was tested for function in the apicomplexan parasite, Toxoplasma gondii, and shown to direct the secretion of the protein. PMID- 15537088 TI - Assessing the effectiveness of coding and non-coding regions in antisense ribosome inhibition of gene expression in Tetrahymena. AB - In Tetrahymena thermophila, an "antisense ribosome" technology has been developed for inhibiting gene expression and generating novel mutants. Short segments of genes are inserted in antisense orientation into an rDNA vector in a region corresponding to an external loop of the folded rRNA. DNA segments derived from the 5'-ends of genes have proven most effective in reducing cognate gene expression. To investigate the efficacy of other genic regions, we generated Tetrahymena cell lines with antisense ribosome constructs containing 100-bp DNA segments derived from the 5'-ends, 3'-ends, and internal coding regions of two non-essential genes, granule lattice protein 1 and macronuclear histone H1. The 5'- and 3'-end constructs inhibited gene expression, but antisense ribosomes derived exclusively from coding regions had little effect. PMID- 15537089 TI - Identification of amoebae implicated in the life cycle of Pfiesteria and Pfiesteria-like dinoflagellates. AB - This study was undertaken to assess whether amoebae commonly found in mesohaline environments are in fact stages in the life cycles of Pfiesteria and Pfiesteria like dinoflagellates. Primary isolations of amoebae and dinoflagellates were made from water and sediment samples from five tributaries of the Chesapeake Bay. Additional amoebae were also cloned from bioassay aquaria where fish mortality was attributed to Pfiesteria. Electron microscopy and small subunit (SSU) rRNA gene sequence analysis of these isolates clearly demonstrated that the commonly depicted amoeboid form of Pfiisteria is very likely a species of Korotnevella and is unrelated to Pfiesteria or Pfiesteria-like dinoflagellates. We have determined that the Pfiesteria and Pfiesteria-like dinoflagellates examined in this study undergo a typical homothallic life cycle without amoeboid stages. Furthermore, we have demonstrated that cloned amoebae sharing morphological characteristics described for stages in the life cycle of Pfiesteria do not transform into dinozoites. The strict clonal isolation and cultivation techniques used in this study substantially support the conclusion that the amoebae and some of the flagellates depicted in the life cycle of Pfiesteria are environmental contaminants of the Pfiesteria culture system and that the Ambush Predator Hypothesis needs to be rigorously reevaluated. PMID- 15537090 TI - Irreversible UV inactivation of Cryptosporidium spp. despite the presence of UV repair genes. AB - Ultraviolet light is being considered as a disinfectant by the water industry because it appears to be very effective for inactivating pathogens, including Cryptosporidium parvum. However, many organisms have mechanisms for repairing ultraviolet light-induced DNA damage, which may limit the utility of this disinfection technology. Inactivation of C. parvum was assessed by measuring infectivity in cells of the human ileocecal adenocarcinoma HCT-8 cell line, with an assay targeting a heat shock protein gene and using a reverse transcriptase polymerase chain reaction to detect infections. Oocysts of five different isolates displayed similar sensitivity to ultraviolet light. An average dosage of 7.6 mJ/cm2 resulted in 99.9% inactivation, providing the first evidence that multiple isolates of C. parvum are equally sensitive to ultraviolet disinfection. Irradiated oocysts were unable to regain pre-irradiation levels of infectivity, following exposure to a broad array of potential repair conditions, such as prolonged incubation, pre-infection excystation triggers, and post-ultraviolet holding periods. A combination of data-mining and sequencing was used to identify genes for all of the major components of a nucleotide excision repair complex in C. parvum and Cryptosporidium hominis. The average similarity between the two organisms for the various genes was 96.4% (range, 92-98%). Thus, while Cryptosporidum spp. may have the potential to repair ultraviolet light-induced damage, oocyst reactivation will not occur under the standard conditions used for storage and distribution of treated drinking water. PMID- 15537092 TI - Infection of mouse dermal fibroblasts by the monoxenous trypanosomatid protozoa Crithidia deanei and Herpetomonas roitmani. AB - Traditionally, monoxenous trypanosomatid protozoa are not believed to infect vertebrate cells. Using light and electron microscopy, we show that the monoxenous trypanosomatids Crithidia deanei and Herpetomonas roitmani are able to infect dermal mouse fibroblasts in vitro. We present experimental evidence of phagocytosis of these trypanosomatids, and demonstrate their survival in vertebrate cells. This paper raises the question about the role of C. deanei and H. roitmani, and perhaps other monoxenous trypanosomatid species, in opportunistic infections of immunocompromised individuals and cutaneos lesions in vertebrate hosts. PMID- 15537091 TI - Mixotrophy in the phototrophic harmful alga Cochlodinium polykrikoides (Dinophycean): prey species, the effects of prey concentration, and grazing impact. AB - We first reported here that the harmful alga Cochlodinium polykrikoides, which had been previously known as an autotrophic dinoflagellate, was a mixotrophic species. We investigated the kinds of prey species and the effects of the prey concentration on the growth and ingestion rates of C. polykrikoides when feeding on an unidentified cryptophyte species (Equivalent Spherical Diameter, ESD = 5.6 microm). We also calculated grazing coefficients by combining field data on abundances of C. polykrikoides and co-occurring cryptophytes with laboratory data on ingestion rates obtained in the present study. Cocholdinium polykrikoides fed on prey cells by engulfing the prey through the sulcus. Among the phytoplankton prey offered, C. polykrikoides ingested small phytoplankton species that had ESD's < or = 11 microm (e.g. the prymnesiophyte Isochrysis galbana, an unidentified cryptophyte, the cryptophyte Rhodomonas salina, the raphidophyte Heterosigma akashiwo, and the dinoflagellate Amphidinium carterae). It did not feed on larger phytoplankton species that had ESD's > or = 12 microm (e.g. the dinoflagellates Heterocapsa triquetra, Prorocentrum minimum, Scrippsiella sp., Alexandrium tamarense, Prorocentrum micans, Gymnodinium catenatum, Akashiwo sanguinea, and Lingulodinium polyedrum). Specific growth rates of C. polykrikoides on a cryptophyte increased with increasing mean prey concentration, with saturation at a mean prey concentration of approximately 270 ng C ml(-1) (i.e. 15,900 cells ml(-1)). The maximum specific growth rate (mixotrophic growth) of C. polykrikoides on a cryptophyte was 0.324 d(-1), under a 14:10 h light-dark cycle of 50 microE m(-2) s(-1), while its growth rate (phototrophic growth) under the same light conditions without added prey was 0.166 d(-1). Maximum ingestion and clearance rates of C. polykrikoides on a cryptophyte were 0.16 ng C grazer( 1)d(-1) (9.4 cells grazer(-1)d(-1)) and 0.33 microl grazer(-1)h(-1), respectively. Calculated grazing coefficients by C. polykrikoides on cryptophytes were 0.001-0.745 h(-1) (i.e. 0.1-53% of cryptophyte populations were removed by a C. polykrikoides population in 1 h). The results of the present study suggest that C. polykrikoides sometimes has a considerable grazing impact on populations of cryptophytes. PMID- 15537093 TI - Molecular phylogenetic analysis places Percolomonas cosmopolitus within Heterolobosea: evolutionary implications. AB - Percolomonas cosmopolitus is a common free-living flagellate of uncertain phylogenetic position that was placed within the Heterolobosea on the basis of ultrastructure studies. To test the relationship between Percolomonas and Heterolobosea, we analysed the primary structure of the actin and small-subunit ribosomal RNA (SSU rRNA) genes of P. cosmopolitus as well as the predicted secondary structure of the SSU rRNA. Percolomonas shares common secondary structure patterns of the SSU rRNA with heterolobosean taxa, which, together with the results of actin gene analysis, confirms that it is closely related to Heterolobosea. Phylogenetic reconstructions based on the sequences of the SSU rRNA gene suggest Percolomonas belongs to the family Vahlkampfiidae. The first Bayesian analysis of a large taxon sampling of heterolobosean SSU rRNA genes clarifies the phylogenetic relationships within this group. PMID- 15537094 TI - Soil ciliate species richness and abundance associated with the rhizosphere of different subtropical plant species. AB - Soil protozoa, and ciliates in particular, represent a microbial group abundant in the rhizosphere with an influential role on nutrient cycling. Under laboratory conditions, ciliates regulate the size and the composition of bacterial communities, and appear to stimulate ammonification and nitrification. In spite of their important ecological role, our understanding about the factors that control their diversity and abundance in natural forest ecosystems is still rudimentary. Plant species-specific interactions have been demonstrated between plants and soil bacteria and mycorrhizal fungi, due in part to the release of phytohormones and C- and N-rich exudates. We tested the hypothesis that the rhizosphere environments of different plant species also influence the species richness and abundance of soil ciliates. Plant effect, soil pH, moisture content, microbial biomass C, and inorganic nitrogen were measured among five plant species to determine the best predictor variables for soil ciliate species richness and total abundance in a subtropical moist forest in Puerto Rico. Based on an analysis of variance, we rejected the hypothesis that there was a plant species-specific effect on soil ciliates, unlike other microbial groups mentioned above. Using multiple regression analysis, we demonstrated that the flush of total inorganic nitrogen was the best predictor variable for both species richness and abundance of ciliates. PMID- 15537095 TI - [Laws (patterns) and principles of the Armed Forces medical support]. AB - The modern ideas of methodological, philosophic and subject aspects of military medicine laws and troops medical support principles are described. Periodization of occurrence and application of the Russian Armed Forces medical support organizational principles is presented. The classification of organizational principles according to 5 signs is given. PMID- 15537096 TI - [The peculiarities of treatment-and-evacuation measures in the troops during military operations conducted in the hilly country]. PMID- 15537097 TI - [Modern ideas about the mechanisms of gunshot injury formation (Report 2)]. AB - The results of experimental study of circulatory acute phase response to the gunshot trauma have shown that triggering of pathologic cascade defined as the traumatic shock is attributed to the free radical concentration created by the wounding projectile in the damaged tissues due to injury sphere formation. So the further study of etiology and pathogenesis of combat pathology is required, the results of which will permit to improve considerably the quality of medical aid rendered to the wounded and casualties. PMID- 15537098 TI - [The implantable cardioverter-defibruillators used in the prophylaxis of sudden cardiac death]. PMID- 15537099 TI - [Influence of cortexin on memory and attention]. AB - The authors present the review of works devoted to one of peptide bioregulators (the biologically active agent) known as cortexin, which is used in memory and attention recovery. The cerebral cortex drug cortexin was created in Military Medical Academy in 1986. The cortexin favourable effect on cerebral mnestic and cognitive functions in different patient groups was noted. The possible mechanisms involved in recovery of these higher nervous activity functions are discussed. PMID- 15537100 TI - [Andrological aspects of varicocele surgery in teenagers]. PMID- 15537101 TI - [Arterial hypertension and cerebrovascular disorders in veterans belonging to special risk subunits]. PMID- 15537102 TI - [Arbidol used in the prophylaxis of acute respiratory viral infections and their complications in servicemen]. AB - The prophylactic action of arbidol to prevent the acute respiratory viral infections and their complications (extra-hospital pneumonia) was studied under conditions of two military collectives during winter and summer time. The data obtained confirm the prophylactic activity of the drug in respect of ARVI. Regardless of the degree of disease epidemic rise among the servicemen who didn't take arbitol the minimal threshold of grippe and other ARVI incidence (10-15%) remained in the experimental group. The incidence of pneumonia decreased. It was connected with decrease in viral-and-bacterial pneumonia. The number of patients with bacterial (generally pneumococcal) pneumonia didn't change. PMID- 15537103 TI - [Age-related changes in operators' activities and brain bioelectrical activity in the Navy specialists]. AB - Psycho- and neurophysiologic investigation of 66 Navy specialists aged 18-71 has revealed the consecutive age-related worsening of different components of operators' activities with predominant and more early disorders in the processes of sensomotor coordination and attention. Apparently the age-related changes in functional state of certain cerebral areas underlie the observed disorders in operators' activities. It is supported by the results of correlation analysis between neurophysiologic and psychophysiologic indices. PMID- 15537104 TI - [The department of medical service organization and tactics of the S M Kirov Military Medical Academy is 75 years]. PMID- 15537105 TI - [An outstanding figure of military and civil health services]. PMID- 15537106 TI - [The military rest homes (70 years since the day of creation)]. PMID- 15537107 TI - NSF gives 'green light' to improve services. PMID- 15537108 TI - The journey to accepting support: how parents of profoundly disabled children experience support in their lives. AB - Advances in medical knowledge and care have extended the lives of children with profound and multiple disabilities. In most cases it is the parents who meet the often complex and continual needs of their child with disabilities in their own home. This study explored the experience of support in the lives of such parents. The interpretive, hermeneutic phenomenology of Heidegger was employed to create a detailed and authentic account of the parents' experiences of support. Five interrelated themes emerged from data from in-depth interviews with six parents randomly selected from a purposive sample in a special school setting. The themes were: parents' feelings about support, the journey to accepting support, support as a loss, disability and the parent and the supportive relationship. Understanding the experience of support from the parent's perspective may lead to a consideration of flexible systems that challenge practice to ensure that supporters listen, learn, develop and deliver support in ways that are helpful. PMID- 15537109 TI - Early warning tools to identify children at risk of deterioration: a discussion. PMID- 15537110 TI - Statistics made simple. Part 4. Choosing the right statistical test. PMID- 15537111 TI - Smacking: a family perspective. PMID- 15537112 TI - 'Children are unbeatable' a nurse's perspective. PMID- 15537113 TI - The NSF for children, young people and maternity services: an overview. AB - Unlike many of the other national service frameworks for England which are disease or condition specific, the children's national service framework encompasses all aspects of children's and young people's health care. The other major difference is that standards do not have detailed targets with set timescales but centre upon principles and examples of good practice. The overriding principle is to ensure quality and safety of care. The implementation of the NSF will form part of the wider programme aimed at improving standards and outcomes for all children. The publication Every Child Matters (DfES 2003) sets out a vision of the outcomes and commitment to support every child to: be healthy, stay safe, enjoy and achieve, make a positive contribution, achieve economic well-being. The NSF standards will feature within the new integrated inspection framework around children's services. Local champions will be crucial to ensuring that the standards are fully implemented at local level. Nurses, midwives and health visitors should take every opportunity to influence developments and ensure that they become actively involved in local discussions and decision-making forums. PMID- 15537114 TI - 'Now wash your hands please': teaching health concepts to very young children. PMID- 15537115 TI - Culture, ethnicity, race: what's the difference? AB - Culture can be seen as a dynamic, ever-changing process. When caring for children and families from diverse backgrounds nurses need to be able to accomodate differences willingly and competently. Do not be afraid to ask--you cannot know everything about someone else's culture. Every unit/team should have contact details of advocates, translators, and local leaders whose support the family may desire, and have information available in other languages which explain the terminology used by healthcare professionals. Nurses can show respect by being available and accessible and by not imposing their own beliefs or agendas. PMID- 15537116 TI - Was hospital liable for sexual assault of female pt. by male nurse? PMID- 15537117 TI - Nurse's expert opinion on 'causation' o.k. for pre suit purpose. Case on point: Apostolico v Orlando Regional Health Care System, 871 So.2d 283--FL (2004). PMID- 15537118 TI - KY: Nurse killed in crash of employer's helicopter: was workers' compensation 'exclusive' remedy? PMID- 15537119 TI - Did medical board exceed authority in limiting CRNA privileges? Case on point: Ortiz v. Dept. of Health, Board of Medicine, 2004 WL 1621440 So.2d--FL. PMID- 15537120 TI - Pulmonary embolism. AB - Pulmonary embolism is a commonly suspected but underdiagnosed condition of clinical significance. Preventable deaths continue to occur. We begin this article with an overview of prognosis, clinical evidence, signs and symptoms, and risk factors, followed by an in-depth evaluation of diagnostic techniques and treatment modalities. The greatest improvement in mortality from pulmonary embolism is likely to come from improved and aggressive prevention and prophylaxis by the critical care team. PMID- 15537121 TI - Adult respiratory distress syndrome. AB - ARDS or acute respiratory distress syndrome continues to be a considerable critical care challenge. Mortality has not decreased significantly over the last more than 30 years. This article presents an overview of origin, evaluation, and treatment of ARDS. Recent findings relative to onset and precipitators of ARDS have led to changes in evaluation and treatment plans. Clinical and radiologic descriptors in assessment of the patient with ARDS are discussed. Ventilatory modes and nursing interventions to optimize patient outcomes are identified. The challenges of outcomes issues presented offer opportunities for further study. PMID- 15537122 TI - Acute exacerbation of COPD: nursing application of evidence-based guidelines. AB - Nurses in acute care settings play a vital role in caring for individuals during an acute exacerbation of chronic obstructive pulmonary disease (COPD), the fourth leading cause of death in the United States. In addressing this health concern, the Global Initiative for Chronic Obstructive Lung Disease Report summarized the goals for COPD management and recommended treatment supported by current data and research. It is imperative that our clinical nursing practice is based upon research-supported interventions: use of appropriate medications, monitoring acid base status, administering controlled oxygen therapy, assessing the need for mechanical ventilation, and close monitoring of comorbid illnesses. Health promotion includes patient and family education on early recognition of symptoms, smoking cessation strategies, and participation in pulmonary rehabilitation that can reduce long-term morbidity from this chronic disease. PMID- 15537123 TI - Respiratory failure: an overview. AB - Respiratory failure is defined as a failure in gas exchange due to an impaired respiratory system--either pump or lung failure, or both. The hallmark of respiratory failure is impairment in arterial blood gases. This review describes the mechanisms leading to respiratory failure, the indices that can be used to better describe gas exchange abnormalities and the physiologic and clinical consequences of these abnormalities. The possible causes of respiratory failure are then briefly mentioned and a quick reference to the clinical evaluation of such patients is made. Finally treatment options are briefly outlined for both acute and chronic respiratory failure. PMID- 15537124 TI - IT matters, but ideas matter more. PMID- 15537125 TI - New stuff. PMID- 15537126 TI - Information security strategies for healthcare--defining the roadmap: part 3. PMID- 15537127 TI - JHIM quick study: clinical decision support systems. PMID- 15537128 TI - Physicians and digital medicine: some thoughts on the future. PMID- 15537129 TI - Beating the system--pitfalls of bar code medication administration. AB - Beating the system to maximize efficiency will yield no positive return on a bar code investment, in terms of reducing medical errors. The critical success factor in the bar coding scenario is that of operational effectiveness. By shifting the nurses' focus from that of efficiency to that of operational effectiveness, the use of bar coding medication technology will reduce medication errors. CIOs looking for a quick fix to reducing medication errors through the use of bar coding technology need to understand that the technology alone won't solve their problem. Rather, it's the balance between operational efficiency and effectiveness among multiple departments working together, both vertically and horizontally, which will yield the greatest ROI. PMID- 15537130 TI - The power of picture archiving and communication systems: strategic hospital considerations. AB - As healthcare information becomes more digitized, hospitals are adopting critical diagnostic tools to improve patient care decisions, while reducing errors and increasing overall efficiency. According to Radiology (2000), 75 percent of the imaging procedures performed in 1995 used devices that were not available in 1970. Today, imaging applications continue to move from research and development into common use each year With regard to digital imaging, Picture Archiving and Communication Systems (PACS) perhaps have changed radiology more than any other application in the past few decades. Once regarded as "nice-to-have-but-too expensive" devices, PACS are rapidly moving into the mainstream of "must-have" solutions for quality-conscious and forward-thinking hospital CEOs. However, before widespread implementation of PACS can occur, several critical factors should be taken into consideration. PMID- 15537131 TI - The Office of Advanced Technology: providing focus on the piloting and implementation of new technology. AB - Rarely does an organization have the luxury of employing full-time staff whose sole purpose is to seek out, select, pilot, and recommend new technology. If such an effort is not a dedicated activity, an organization will have a hard time keeping abreast of technological development with an eye towards successful deployment to the benefit of the business. This paper summarizes the role and activities of the Office of Advanced Technology (OAT) at the Mayo Clinic in Scottsdale, AZ. The OAT has been in existence since 2001 and is staffed on a part time basis by three individuals: a member of the physician consulting staff, a nurse informaticist, and a member of the allied health Information Technology staff. The office has successfully conducted pilot projects in several new technology areas, advocating the adoption of some technologies and deferring on others. Moreover, they have been instrumental serving as a spokesperson and single point of contact for all new technology initiatives of this multi specialty practice. PMID- 15537132 TI - RFID and bar codes--critical importance in enhancing safe patient care. AB - Medication administration recording and supply management are complex and interrelated processes. The integration of bar codes and radio frequency identification tags are viewed as critical in achieving effective and safe patient care. However, these systems are complex; all parts need to be aligned, and the systems must work together to produce the desired outcomes. In healthcare, automation using bar coding and RFID capabilities is of growing importance because of the Institutes of Medicine study and the integrated electronic medical record. Healthcare systems today are increasingly complex, and while bar codes and RFID technologies provide opportunities for enhanced patient care, systems using these capabilities must be carefully planned to achieve optimal outcomes. PMID- 15537133 TI - Adding user-friendliness and ease of implementation to continuous speech recognition technology with speech macros: case studies. AB - Continuous Speech Recognition Technology implementation is expensive, and the failure of leading companies in this niche can hamper usefulness. C-SRT, if deployed and used with speech macros, experiences vastly improved implementations and drastically reduces medical transcription costs.A speech macro is a short phrase that is automatically translated into a block of text or a graphic display. A more powerful form of speech macro can bring up predefined templates and insert spoken text into the proper position automatically, based on its interpretation. Cases from the author's consulting experiences and from medical journals emphasize the need for speech macros from a cost-benefit standpoint. A prototype program is introduced that facilitates the process of creating macros. The need for macro management software is reconciled with current research on speech recognition and technology adoption. A planned experiment is discussed. PMID- 15537134 TI - The application of volume-outcome contouring in data warehousing. AB - Despite a compelling body of published research on the nature of provider volume and clinical outcomes, healthcare executives and policymakers have not managed to develop and implement systems that are useful in directing patients to higher volume providers via selective referral or avoidance. A specialized data warehouse application, utilizing hospital discharge data linked to physician biographical information, allows detailed analysis of physician and hospital volume and the resulting pattern (contour) of related outcomes such as mortality, complications, and medical errors. The approach utilizes a historical repository of hospital discharge data in which the outcomes of interest, important patient characteristics and risk factors used in severity-adjusting of the outcomes are derived from the coding structure of the data. PMID- 15537135 TI - Electronic implementation of national nursing standards--NANDA, NOC and NIC as an effective teaching tool. AB - With the increased interest in evidence-based medicine, Internet access and the growing emphasis on national standards, there is an increased challenge for teaching institutions and nursing services to teach and implement standards. At the same time, electronic clinical documentation tools have started to become a common format for recording nursing notes. The major aim of this paper is to ascertain and assess the availability of clinical nursing tools based on the NANDA, NOC and NIC standards. Faculty at 20 large nursing schools and directors of nursing at 20 hospitals were interviewed regarding the use of nursing standards in clinical documentation packages, not only for teaching purposes but also for use in hospital-based systems to ensure patient safety. A survey tool was utilized that covered questions regarding what nursing standards are being taught in the nursing schools, what standards are encouraged by the hospitals, and teaching initiatives that include clinical documentation tools. Information was collected on how utilizing these standards in a clinical or hospital setting can improve the overall quality of care. Analysis included univariate and bivariate analysis. The consensus between both groups was that the NANDA, NOC and NIC national standards are the most widely taught and utilized. In addition, a training initiative was identified within a large university where a clinical documentation system based on these standards was developed utilizing handheld devices. PMID- 15537136 TI - The case for regulatory compliance of electronic medical records about human tissue intended for transplantation. AB - It is vital that patients receiving human tissue by transplantation be able to trust in the suitability of the product. In short, patient safety must be ensured to the highest degree possible. To this end, firms operating tissue banks and those providing tissue donor coordination services are inspected by the U.S. Food and Drug Administration (FDA). The suitability of tissue for transplantation is objectively documented at all phases, beginning with donor screening. This process often involves the use of custom software applications that produce electronic medical records (EMRs) and databases that store the results. When a firm elects to keep the medical records relevant to human tissue products in electronic form, the record system must comply with applicable codified regulations. Unfortunately for the patient, many firms have yet to achieve regulatory compliance, and FDA enforcement is only beginning to approach the intensity appropriate to regulate this fast-growing industry. This article discusses current FDA trends in enforcement and rule-making related to human tissue intended for transplantation, as well as expectations the FDA has during an inspection. PMID- 15537137 TI - Patient Education and Recovery Learning System (PEARLS) pathway: a tool to drive patient centered evidence-based practice. AB - Myocardial infarction is a significant problem for Americans and individuals in developed countries. According to the American Heart Association, coronary heart disease is the single largest killer of Americans. Variation in practice is recognized as a key barrier to promoting positive outcomes in patients with cardiac illness. Implementation of clinical practice guidelines has been identified as one way to promote evidence-based practice, reduce practice variation, and ultimately improve the quality of care and outcomes of cardiac patients. Improving patient knowledge and understanding of guideline recommendations and the ways in which evidence-based recommendations are linked to recovery is recognized as one means of facilitating guideline adherence. Automated patient pathways are proposed as a means to improve patient access to guideline-based information at the point of care. Within this context, the prototype development of a Patient Education And Recovery Learning System, or PEARLS, is discussed along with the potential benefits of the use of automated patient pathways as one means for overcoming challenges associated with acute care patient education and facilitating patient adherence to clinical practice guideline recommendations. PMID- 15537138 TI - Applications of ecological interface design in supporting the nursing process. AB - Today's nursing environment is complex, with many sources of data that are often poorly displayed. Ecological interface design (EID) is a systematic approach to designing interfaces to complex systems. EID has been used to design interfaces for aviation displays, power plant monitoring and control, human hemodynamic monitoring, anesthesia monitoring, and neonatal intensive care monitoring and diagnosis. EID makes critical relationships easily visible, eliminating the mental workload of integrating, calculating, or remembering multiple values. This paper reviews past experimental studies of EID in healthcare applications and discusses the application of EID to a decision-support tool for diabetic patients using personal digital assistants. The authors also discuss other contributions that EID could make to the nursing process in the areas of physiological monitoring, decision support, database design, and the measurement and analysis of nurse-sensitive outcomes, including patient safety outcomes. PMID- 15537139 TI - Need for more evidence on vaccination. PMID- 15537141 TI - Is veterinary education too broad? PMID- 15537140 TI - Preparing for a new Veterinary Surgeons Act: making sure the objectives are clear. PMID- 15537142 TI - Prevalence of BSE in cattle found dead, euthanased or emergency slaughtered on farms in western France in 2000, 2001 and 2002. AB - The overall trend and the trend within birth cohorts of the prevalence of bovine spongiform encephalopathy (BSE) in cattle found dead, euthanased or emergency slaughtered on farms in the Bretagne, Basse Normandie and Pays de la Loire regions of France, during the periods from August 7 to December 22 in 2000, 2001 and 2002, were analysed by non-conditional logistic regression, adjusted for the region and for the type of animals. The overall prevalence of BSE during these three periods decreased from 2.71 per 1000 in 2000 to 1.41 per 1000 in 2001 and 0.42 per 1000 in 2002. The prevalence within birth cohorts started to decrease for the cohort born between July 1, 1995 and June 30, 1996 (cohort 95/96) and the trend was reinforced for cohort 96/97, suggesting that the exposure of animals to the BSE agent had started to decrease for animals born after July 1995, that is, one year before the ban on specified risk materials in meat and bone meal was implemented in France. However, considering that most of the animals would have been infected at between six and 18 months of age, the decrease may have been due, at least partly, to this control measure. PMID- 15537143 TI - Septic tenosynovitis of the tarsal sheath of an Arab gelding and suspected sepsis of the lateral digital flexor tendon subsequent to bacterial peritonitis. AB - A 21-year-old Arab gelding with clinical signs of acute peritonitis had a perforating ulcer on the mesenteric border of the jejunum which resulted in localised contamination of the abdomen with ingesta. The affected segment of jejunum was resected and the abdomen was lavaged extensively. Postoperatively, the gelding was treated with broad-spectrum antibiotics, non-steroidal anti inflammatory drugs and intravenous fluids, but after four days it became acutely non-weight bearing on its right hindlimb, and a tendonitis of the lateral digital flexor tendon within the tarsal sheath was identified ultrasonographically. The septic tendonitis was treated with broad-spectrum antibiotics but progressed proximodistally within the tendon until it involved both the tarsal sheath and the associated tarsocrural joint, necessitating the euthanasia of the gelding. PMID- 15537144 TI - Prevalence of porcine circovirus type 2 in aborted fetuses and stillborn piglets. AB - A retrospective study of natural cases of abortion, recorded between October 2000 and September 2002, was made to determine the prevalence of abortions associated with porcine circovirus type 2 (PCV-2). The virus was detected by PCR, immunohistochemistry and in situ hybridisation. A total of 46 (13.1 per cent) of 350 aborted fetuses and stillborn piglets were positive for PCV-2 by PCR, and the virus was detected in fetuses at all stages of gestation. Viral antigen was detected in macrophages from the aborted fetuses and stillborn piglets by immunohistochemistry, and viral DNA was detected by in situ hybridisation. PMID- 15537145 TI - Postal questionnaire survey of scrapie in sheep flocks in Ireland. PMID- 15537146 TI - Clinical findings in a Swiss Braunvieh heifer with a cerebellar abscess. PMID- 15537147 TI - Tracing the regional source of rabies infection in an Israeli dog by viral analysis. PMID- 15537148 TI - Squamous cell carcinoma of the frontal sinus and nasal conchae in an addax (Addax nasomaculatus). PMID- 15537149 TI - Spontaneous parthenogenesis and development of camel (Camelus dromedarius) oocytes. PMID- 15537150 TI - Standards for international trade. PMID- 15537151 TI - Total elbow replacement in dogs. PMID- 15537152 TI - Bendiocarb poisoning in a green woodpecker. PMID- 15537153 TI - Genetic influence on the expression of PCV disease. PMID- 15537154 TI - The chemistry of exploding ants, Camponotus spp. (cylindricus complex). AB - A detailed comparative analysis of the exocrine chemistry of nine Bruneian Camponotus species in the cylindricus complex is reported. Workers of these species are known to have hypertrophied mandibular glands and release their glandular contents suicidally from the head by rupturing the intersegmental membrane of the gaster. All of the species produce mixtures of polyacetate derived aromatics, including hydroxyacetophenones, which display pH-dependent color changes, and aliphatic hydrocarbons and alcohols. In addition, three species contained (6R)-2,6-dimethyl-(2E)-octen-1,8-dioic acid (9) or (3S)-8 hydroxycitro-nellic acid (10a), previously unreported from insects. These compounds were characterized from their spectral data, and confirmed by comparison with synthetic samples. The allomonal role of these compounds is based on numerous field observations, and their chemotaxonomic value is presented. PMID- 15537155 TI - Male-produced aggregation pheromone of the cerambycid beetle Neoclytus acuminatus acuminatus. AB - This is the first fully verified report of an aggregation pheromone produced by a cerambycid beetle species. Field bioassays with adult Neoclytus acuminatus acuminatus (F.) (Coleoptera: Cerambycidae) revealed that males produce a pheromone that attracts both sexes. Extracts of odors from males contained a single major male-specific compound, (2S,3S)-hexanediol. Field trials determined that both sexes were attracted by the racemic blend of (2S,3S)- and (2R,3R) hexanediols and that activity was similar to enantiomerically enriched (2S,3S) hexanediol (e.e. 80.2%). However, the blend of all four 2,3-hexanediol stereoisomers attracted few beetles, indicating inhibition by one or both of the (2R*,3S*)-stereoisomers. Females of the cerambycid Curius dentatus Newman were attracted to traps baited with the four component blend, suggesting that a male produced sex pheromone for this species may contain (2R,3S)-hexanediol and/or (2S,3R)-hexanediol. The pheromone of N. a. acuminatus, and presumed pheromone of C. dentatus, bear structural similarities to those produced by males of six other species in the Cerambycinae (straight chains of 6, 8, or 10 carbons with hydroxyl or carbonyl groups at C2 and C3). It is likely that males of other species in this large subfamily produce pheromones that are variations on this structural motif. PMID- 15537156 TI - Investigation of long-range female sex pheromone of the European tarnished plant bug, Lygus rugulipennis: chemical, electrophysiological, and field studies. AB - The European tarnished plant bug, Lygus rugulipennis, is an important pest of agricultural and horticultural crops throughout Europe. Adult male L. rugulipennis were previously shown to be attracted to traps baited with live virgin females, which suggests the females produce a sex pheromone. Volatiles produced by virgin female L. rugulipennis were shown to contain three components, hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal which elicited electroantennographic (EAG) responses from males in analyses by linked gas chromatography-electroantennography (GC-EAG). They were produced in 1.5:1:0.08 ratio, respectively, by single females. Collections from 1, 2, or 4 virgin females showed the proportions of hexyl butyrate and (E)-4-oxo-2-hexenal to increase relative to that of (E)-2-hexenyl butyrate with increasing number of females. Although these compounds were found in body extracts of both male and female L. rugulipennis, they were not detected in volatiles released by virgin males. EAG dose-response studies showed that both males and females responded to these chemicals with minimal differences in sensitivity between the sexes or to the three components, except that males were more responsive than females to (E) 4-oxo-2-hexenal at the two highest doses tested. Release rates of the compounds from rubber septa, polyethylene vials, and polyethylene sachets were measured under laboratory conditions. Four field tests were carried out using sticky traps baited with all possible binary and tertiary combinations of the three chemicals using different combinations of dispensing systems. Catches of male L. rugulipennis in baited traps were similar to those in unbaited traps. Significantly fewer females were caught on traps baited with blends containing hexyl butyrate than on traps without hexyl butyrate or unbaited traps in one test and overall. The roles of the three compounds and possible reasons for their failure to attract males are discussed. PMID- 15537157 TI - Identification of a sex pheromone component for the blueberry leafminer, Caloptilia porphyretica. AB - Coupled gas chromatographic-electroantennographic detection (GC-EAD) of both gland extracts and effluvial collections from female blueberry leafminer, Caloptilia porphyretica Braun (Lepidoptera: Gracillariidae), showed that females produced a single EAD-active compound. The amount of the compound collected from virgin female C. porphyretica was below GC and mass spectrometry (MS) detection thresholds, even with highly concentrated gland extracts (approximately 150 female equivalent). (E)-11-Hexadecenal (E11-16:Ald) was determined to be a sex pheromone component mainly by comparison of retention times with authentic standards on both polar and nonpolar capillary columns, microreaction-GC-EAD analyses, and field trapping tests. GC-EAD experiments showed that synthetic E11 16:Ald exhibited extraordinarily high electrophysiological activity, stimulating significant male antennal responses at as low as 10 fg. Traps baited with E11 16:Ald alone were attractive to males. Addition of 1 or 3% of its geometric isomer, Z11-16:Ald, to E11-16:Ald did not significantly increase trap captures, but an inhibitory effect was observed at the 10% level. The influence of two kinds of rubber septa on attraction was also evaluated. Male moth captures were higher in traps baited with red rubber septa than with gray rubber septa at 30 300-microg doses. Monitoring of adult flight activity with 3-microg doses of E11 16:Ald indicated at least three distinct flight periods throughout the 2003 season. PMID- 15537158 TI - Behavioral response of Lygus hesperus to conspecifics and headspace volatiles of alfalfa in a Y-tube olfactometer. AB - The western tarnished plant bug, Lygus hesperus Knight, feeds and develops on a variety of weeds in the spring, with later generations moving to alfalfa and cotton where severe damage to reproductive structures can occur. A synthetic attractant for monitoring or mass-trapping L. hesperus, or the identification of potential attractants for natural enemies, would be useful tools for integrated pest management programs. Studies investigated the response of naive and experienced fifth-instar and adult L. hesperus to odors associated with conspecifics and alfalfa, Medicago sativa L. Fifth-instar L. hesperus responded to all plant/insect combinations, whereas female L. hesperus only responded preferentially to vegetative and flowering alfalfa where conspecifics had fed for 24-72 hr, and to vegetative alfalfa where conspecifics were added approximately 30 min before the test began. Males were not attracted to headspace volatiles from any of the alfalfa treatments. Analysis of headspace volatiles showed that (E)-2-hexanal, (Z)-3-hexen-l-ol, alpha-pinene, (Z)-3-hexenyl acetate, (E)-2 hexenyl acetate, limonene, (Z)-ocimene, (E)-beta-ocimene, linalool, (3E)-4,8 dimethyl-1,3,7-nonatriene, and (E, E)-alpha-farnesene are emitted from both vegetative and flowering alfalfa. Indole and (3E, 7E)-4,8,12-trimethyl-1,3,7,11 tridecatetraene were only detected in flowering alfalfa. Damage to alfalfa by L. hesperus increased emissions of (Z)-ocimene, (E)-beta-ocimene, (E)-beta caryophyllene, and (E, E)-alpha-farnesene, while beta-pinene, myrcene, methyl salicylate, and (3E, 7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene were only detected from damaged plants. Thus, individual or mixtures of these alfalfa volatiles may be useful as attractants for capturing nymphs and adult females of L. hesperus in the field. PMID- 15537159 TI - Detection and discrimination of conspecific scents by the anguid slow-worm Anguis fragilis. AB - We tested the ability of male slow-worms, Anguis fragilis, a limbless anguid lizard with secretive, semifossorial habits, to detect chemical associated with conspecifics by using a T-maze in the laboratory. Male slow-worms discriminated conspecific male and female scent deposits. Males selected the arm with female scent, suggesting that scent deposits may be used to locate potential mates. Also, male slow-worms did not avoid the chemicals of other males, suggesting that they are not territorial. However, males discriminated their own scent from those of other males, and spent more time exploring the arm with other male scent, which suggests that scent marks may bear information that could be used in future intrasexual social contexts. We conclude that discrimination of conspecifics based on scents may be more widespread than previously expected among lizards inhabiting visually restricted environments. PMID- 15537160 TI - Statistical problems encountered in trapping studies of scolytids and associated insects. AB - Traps baited with semiochemicals are often used to investigate the chemical ecology of scolytids and associated insects. One statistical problem frequently encountered in these studies are treatments that catch no insects and, thus, have zero mean and variance, such as blank or control traps. A second problem is the use of multiple comparison procedures that do not control the experimentwise error rate. We conducted a literature survey to determine the frequency of these two statistical problems in Journal of Chemical Ecology for 1990-2002. Simulations were then used to examine the effects of these problems on the validity of multiple comparison procedures. Our results indicate that both statistical problems are common in the literature, and when combined can significantly inflate both the experimentwise and per comparison error rate for multiple comparison procedures. A possible solution to this problem is presented that involves confidence intervals for the treatment means. Options to increase the statistical power of trapping studies are also discussed. PMID- 15537161 TI - Benzoic acid derivatives in a hypogastrurid collembolan: temperature-dependent formation and biological significance as deterrents. AB - Two phenolic acids were identified in the collembolan Ceratophysella denticulata: 3-hydroxy-4,5 dimethoxy benzoic acid and 4-hydroxy-3,5-dimethoxybenzoic acid (syringic acid). These are localized on or in the integument of the springtail, in field-collected animals, in a ratio of 47:100 (v/v). Springtails kept under different temperature regimes showed differences in production and ratio of the benzoic acid derivatives. At 20 degrees C, C. denticulata produced only syringic acid, whereas at 10 degrees C both isomers in a ratio of 100:61 (v/v) were detected. Bioassays with C. denticulata as well as with the specialized collembolan predator Stenus comma (Staphylinidae) were carried out. Staphylinid beetles topically treated with the acids try to clean their mouthparts by rubbing them on the ground significantly more often than do control beetles. Both compounds individually and as a natural mixture have deterrent effects towards the predator S. comma. PMID- 15537162 TI - Avian exocrine secretions. I. Chemical characterization of the volatile fraction of the uropygial secretion of the green woodhoopoe, Phoeniculus purpureus. AB - Using gas chromatography-mass spectrometry in conjunction with auxiliary techniques such as solid phase microextraction and determination of double bond positions by means of dimethyl disulfide derivatization, 45 constituents of the uropygial secretion of the green woodhoopoe, Phoeniculus purpureus, have been identified. The majority of these constituents are long-chain branched and unbranched alkanes, and (Z)-alkenes such as (Z)-9-tricosene, and a number of unidentified wax esters. The more volatile fraction of the secretion contained short-chain fatty acids, aldehydes, aliphatic and heterocyclic aromatic amines, ketones, and dimethyl sulfides. This group of volatile compounds is responsible for the obnoxious odor of the secretion and also for its defensive action against predators. PMID- 15537163 TI - Field-testing of methyl salicylate for recruitment and retention of beneficial insects in grapes and hops. AB - Evidence for recruitment and retention of beneficial insects in grapes and hops using controlled-release dispensers of methyl salicylate (MeSA), a component of herbivore-induced volatile blends, is presented. In a replicated experiment conducted in a juice grape vineyard, sticky cards in blocks baited with MeSA captured significantly greater numbers of five species of predatory insects (Chrysopa nigricornis, Hemerobius sp., Deraeocoris brevis, Stethorus punctum picipes, Orius tristicolor) than unbaited blocks. Four insect families (Syrphidae, Braconidae, Empididae, Sarcophagidae) were also significantly more abundant in the MeSA-baited blocks, as indicated by sticky card captures. Canopy shake samples and sticky card monitoring conducted in a MeSA-baited, unsprayed hop yard indicated development and maintenance of a beneficial arthropod population that was nearly four times greater than that present in an unbaited reference yard. Four times as many S. punctum picipes and six times as many O. tristicolor were sampled in the MeSA yard. Similar contrasts in abundance of these predators and others were apparent when compared with levels recorded in the yard in previous years. The large population of predatory insects in the MeSA baited hop yard was associated with a dramatic reduction in spider mite numbers, the major arthropod pest of hops, in late June, and subeconomic populations were maintained for the rest of the season. The evidence presented here is highly suggestive that the use of controlled-release MeSA in a crop could increase recruitment and residency of populations of certain beneficial insects. This strategy may have the potential to enhance the efficacy and reliability of conservation biological control in crop pest management. PMID- 15537164 TI - Responses of Teretrius nigrescens toward the dust and frass of its prey, Prostephanus truncatus. AB - Teretrius nigrescens is considered to be a specialized predator of Prostephanus truncatus, a serious pest of stored maize and dried cassava roots. Using a bait bag behavioral bioassay, this investigation found T nigrescens to be strongly arrested by dust and frass produced by P. truncatus on maize, whereas responses to maize flour were weak. Attempts to increase the arrestiveness of flour by altering its physical properties (coarseness or particle size range) were unsuccessful. The arrestive property of dust/frass did not degrade with long-term storage nor did it volatilize or degrade with oven baking up to 150 degrees C. However, extraction with methanol and reapplication was successful in transferring the arrestive property from the dust/frass onto maize flour. The prey dust/frass also induced more oviposition than did maize flour, as judged by production of F1 offspring. The results provide strong evidence for the existence of a high boiling point compound(s) in P. truncatus dust/frass that acts as a contact kairomone for T nigrescens. This, or another compound(s) also seems to act as an oviposition stimulant for female T. nigrescens. PMID- 15537165 TI - Identification and quantitation of compounds in a series of allelopathic and non allelopathic rice root exudates. AB - An investigation of the chemical basis for rice allelopathy to the rice weed arrowhead (Sagittaria montevidensis) was undertaken using GC/MS and GC/MS/MS techniques. Twenty-five compounds were isolated and identified from the root exudates of both allelopathic and non-allelopathic rice varieties. Phenolics, phenylalkanoic acids, and indoles were among the chemical classes identified. Two indoles previously unreported in rice were detected in the exudates, 5-hydroxy-2 indolecarboxylic acid and 5-hydroxyindole-3-acetic acid. Several other compounds identified in this study have not previously been reported in rice root exudates, namely mercaptoacetic acid, 4-hydroxyphenylacetic acid, and 4-vinylphenol. The levels of 15 compounds present in the exudates were quantified using GC/MS/MS. Six of the seven most abundant compounds were phenolic acids. Significant differences exist between the allelopathic and non-allelopathic cultivars in their production of three of these six compounds. Greater amounts of trans ferulic acid, p-hydroxybenzoic acid, and caffeic acid were detected in the exudates of allelopathic cultivars. The seventh compound, abietic acid, was significantly higher in the non-allelopathic cultivars. PMID- 15537166 TI - Evaluation of putative allelochemicals in rice root exudates for their role in the suppression of arrowhead root growth. AB - In previous studies, 15 putative allelopathic compounds detected in rice root exudates were quantified by GC/MS/MS. In this study, multiple regression analysis on these compounds determined that five selected phenolics, namely caffeic, p hydroxybenzoic, vanillic, syringic, and p-coumaric acids, from rice exudates were best correlated with the observed allelopathic effect on arrowhead (Sagittaria montevidensis) root growth. Despite this positive association, determination of the phenolic acid dose-response curve established that the amount quantified in the exudates was much lower than the required threshold concentration for arrowhead inhibition. A similar dose-response curve resulted from a combination of all 15 quantified compounds. Significant differences between the amounts of trans-ferulic acid, abietic acid, and an indole also existed between allelopathic and non-allelopathic rice cultivars. The potential roles of these three compounds in rice allelopathy were examined by chemoassay. Overall, neither the addition of trans-ferulic acid nor 5-hydroxyindole-3-acetic acid to the phenolic mix significantly contributed to phytotoxicity, although at higher concentrations, trans-ferulic acid appeared to act antagonistically to the phytotoxic effects of the phenolic mix. The addition of abietic acid also decreased the inhibitory effect of the phenolic mix. These studies indicate that the compounds quantified are not directly responsible for the observed allelopathic response. It is possible that the amount of phenolic acids may be indirectly related to the chemicals finally responsible for the observed allelopathic effect. PMID- 15537167 TI - Modulatory influence of noscapine on the ethanol-altered hepatic biotransformation system enzymes, glutathione content and lipid peroxidation in vivo in rats. AB - The modulatory potential of noscapine, an opium alkaloid was assessed on the ethanol-induced changes in hepatic drug metabolizing enzyme systems, glutathione content and microsomal lipid peroxidation. Noscapine was administered orally to male Wistar rats at a dose level of 200 mg/kg bw alone as well as in combination with 50% ethanol (v/v) for 5 days. Noscapine administration was associated with a approximately 91% decrease in hepatic microsomal cytochrome P-450 content. A decline of approximately 36% was observed in the NADPH-cytochrome c reductase activity on noscapine administration. The lowering of cytochrome P-450 levels on noscapine administration was accompanied by a concomitant increase in heme oxygenase activity as well as serum bilirubin levels. Our results indicate that the combination dosage of noscapine and ethanol antagonised the ethanol-induced elevation of cytochrome P-450 levels. Noscapine fed rats had decreased glutathione (GSH) content and enhanced lipid peroxidation compared to control rats as indexed by MDA method. Further, noscapine and ethanol coexposure produced a more pronounced elevation in lipid peroxidation and the glutathione levels also decreased significantly. We speculate on the basis of our results that the significant enhancement of lipid peroxidation on combination dosage of noscapine and ethanol is a consequence of depletion of glutathione to certain critical levels. The inhibition of glutathione-S-transferase (GST) as well as lowering of cytochrome P-450 suggests that the biotransformation of noscapine and ethanol is significantly altered following acute coexposures. PMID- 15537168 TI - In vitro formation of selegiline-N-oxide as a metabolite of selegiline in human, hamster, mouse, rat, guinea-pig, rabbit and dog. AB - It is well established in the litrature, that selegiline is metabolised to its N dealkylated metabolites, N-desmethylselegiline, methamphetamine and amphetamine. However, most studies on selegiline metabolism did not characterize the species differences in the formation of the metabolites. Therefore, in this study, we investigated the in vitro metabolism of selegiline in liver microsomes of different species. In addition, to the previously well-characterized metabolites, selegiline-N-oxide (selegiline-NO) was found to be formed as a metabolite of selegiline in rat liver microsomal preparation. The results of experiments with liver microsomes from other species indicated species differences in the rate and extent of formation of selegiline-NO. The dog and hamster liver microsomal preparations were the most active in terms of selegiline-NO production, whereas little selegiline was metabolized to its N-oxide in human liver microsomes. When selegiline-NO was incubated with rat liver microsomes, no metabolism occurred. When a short incubation time was applied in selegiline expriments no increase in the amount of selegiline-NO was detected. Accordingly, it was clear that selegiline was not metabolized to the N-dealkylated or N,N-bis-dealkylated compounds via selegiline-NO. Studies with different isoenzyme inhibitors indicated that the formation of selegiline-NO might be catalyzed at least partly by cytochrome P450 (CYP) 2D6 and CYP3A4. With the exception of hamster microsomes in the microsomal preparations in vitro, the formation of the R,S-stereoisomer of selegiline-NO was preferred. PMID- 15537169 TI - The in vitro metabolism of desglymidodrine, an active metabolite of prodrug midodrine by human liver microsomes. AB - The human cytochrome P450 (CYP) isoforms catalyzing the oxidation metabolism of desglymidodrine (DMAE), an active metabolite of midodrine, were studied. Recombinant human CYP2D6, 1A2 and 2C19 exhibited appreciable catalytic activity with respect to the 5'-O-demethylation of DMAE. The O-demethylase activity by the recombinant CYP2D6 was much higher than that of other CYP isoforms. Quinidine (a selective inhibitor of CYP2D6) inhibited the O-demethylation of DMAE in pooled human microsomes by 86%, while selective inhibitors for other forms of CYP did not show any appreciable effect. Although the activity of CYP2D6 was almost negligible in the PM microsomes, the O-demethylase activity of DMAE was found to be maintained by about 25% of the pooled microsomes. Furafylline (a selective inhibitor of CYP1A2) inhibited the M-2 formation in the PM microsomes by 57%. The treatment of pooled microsomes with an antibody against CYP2D6 inhibited the formation of M-2 by about 75%, whereas that of the PM microsomes did not show drastic inhibition. In contrast, the antibody against CYP1A2 suppressed the activity by 40 to 50% in the PM microsomes. These findings suggest that CYP2D6 have the highest catalytic activity of DMAE 5'-O-demethylation in human liver microsomes, followed by CYP1A2 to a small extent. PMID- 15537170 TI - Lipid kinetics in obese patients undergoing laparoscopy. The impact of cortisol inhibition by etomidate. AB - The aim of this study was to investigate the response of cortisol, insulin and lipid parameters [serum Lipoprotein Lipase activity, choleseryl-ester transfer protein, triglycerides, total Cholesterol, High Density Lipoprotein, Free Fatty Acids] during the perioperative period in obese patients undergoing laparoscopic cholecystectomy. Twenty obese patients were included and divided in two groups. In group A (n=10) patients were anaesthetized with propofol and group B (n=10) with etomidate. Blood samples were collected before induction in anaesthesia, just after the end of the operation and at one, two and three hours postoperatively. According to our results, in both groups serum LPL activity showed a significant decrease whereas serum Free Fatty Acids a potent increase over time. Likewise, both groups did not demonstrate significant changes over time in choleseryl-ester transfer protein activity, total cholesterol, triglycerides, High Density Lipoprotein or insulin concentrations in serum. Furthermore, cortisol release was significantly inhibited in the etomidate group while substantially enhanced in propofol group. Additionally, apart of triglycerides, no difference was found between the two groups in all the lipid parameters and insulin concentrations. In conclusion, serum Free Fatty Acids levels and Lipoprotein Lipase activity demonstrated significant alterations in obese patients underwent laparoscopic cholecystectomy and this result did not seem to be related with the anaesthetic agent used for induction in anaesthesia. PMID- 15537171 TI - Alteration of oral salivary pharmacokinetics of paracetamol by an investigational anti-malarial phytomedicine, in healthy human volunteers. AB - Effect of an investigational anti-malarial phytomedicine (AM1) on the salivary pharmacokinetics of paracetamol (PCM), in healthy human volunteers was examined. The drug salivary level of paracetamol was determined using spectrophotometric method. Lower concentrations of PCM was observed in concomitant administration of 1000mg PCM tablets with 250mg AM1 capsule compared with administration of 1000mg paracetamol tablets alone (control). The area under the curve (AUC), time for maximum concentration (t(max)), maximum concentration (Cmax) were generated using the GRAPHPAD PRISM software version 2.0. The pharmacokinetic parameters were calculated from the semi-logarithmic plots of concentration-time data for PCM. The absorption kinetics parameters kab, lag-time, t1/2ab for both test (0.03min( 1), 9min, 18min.) and control (0.03min(-1), 9min, 22.8min.) were identical. There were statistically significant reductions in the bioavailability parameters AUC, Cmax, between control (1856microg/ml/min, 8.08microg/ml) and test (847.87microg/ml/min, 4.59microg/ml/) experiments. There were also insignificant but definite alterations in the elimination phase kinetic parameters kel, t1/2el, between control (0.1172min(-1), 69min.) and test (0.01027min(-1), 75min). The study shows that the botanical anti-malarial remedy (AM1) reduced the systemic availability of PCM when used concomitantly. PMID- 15537172 TI - Pharmacokinetics and tissue distribution of gentiopicroside following oral and intravenous administration in mice. AB - The pharmacokinetics and tissue distribution of Gentiopicroside (GPS), one of the major active components of the Gentiana species of medicinal plants, was studied following oral and intravenous administration in mice. The distribution of GPS in mice after oral and intravenous doses could be fitted to a two-compartments open model. The serum half-life of GPS was 6.1 h and 2.8 h for intravenous and oral administration, respectively. The Tmax of GPS after oral administration was 0.50 h, and the bioavailability was 39.6%. The AUC gradient in individual tissues following intravenous administration was kidney >serum >liver >spleen >lung >thymus >fat >heart >muscle >stomach >intestinal >brain. The MRT gradient was muscle >serum >lung >spleen >lung >intestinal>heart >stomach >brain >liver >thymus >kidney >fat. Overall the data show that GPS could be absorbed rapidly in mice, but with a low bioavailability, and could distribute to tissues extensively, but was generally cleared quickly with short MRTs. The study demonstrates the need for repeated dosage, or better, a slow release formulation as an ideal means of administering GPS. PMID- 15537174 TI - The flu snafu. PMID- 15537173 TI - Classical Michaelis-Menten and system theory approach to modeling metabolite formation kinetics. AB - When single doses of drug are administered and kinetics are linear, techniques, which are based on the compartment approach and the linear system theory approach, in modeling the formation of the metabolite from the parent drug are proposed. Unlike the purpose-specific compartment approach, the methodical, conceptual and computational uniformity in modeling various linear biomedical systems is the dominant characteristic of the linear system approach technology. Saturation of the metabolic reaction results in nonlinear kinetics according to the Michaelis-Menten equation. The two compartment open model with Michaelis Menten elimination kinetics is theorethicaly basic when single doses of drug are administered. To simulate data or to fit real data using this model, one must resort to numerical integration. A biomathematical model for multiple dosage regimen calculations of nonlinear metabolic systems in steady-state and a working example with phenytoin are presented. High correlation between phenytoin steady state serum levels calculated from individual Km and Vmax values in the 15 adult epileptic outpatients and the observed levels at the third adjustment of phenytoin daily dose (r=0.961, p<0.01) were found. PMID- 15537176 TI - Social work readers describe "useful knowledge". PMID- 15537175 TI - Click to get sick? People who get medical advice from the Web actually get worse, a study shows. PMID- 15537177 TI - Comprehensive community initiatives: a rural perspective. AB - This article challenges the notion that the comprehensive community initiative (CCI) is a singularly urban intervention strategy by comparing a rural comprehensive initiative with the literature on urban CCIs. Characteristics of CCIs in urban settings are discussed and compared with Warren Family Institute (WFI), a demonstration project in rural North Carolina. Findings support the thesis that the CCI has been implemented and found effective in rural settings. The author presents a comparison of the common complications faced by urban CCIs with those encountered during the planning, implementation, and evaluation of WFI. Differences between these two CCIs were rooted in demographic, geographic, and sociohistoric distinctions between rural and urban settings. Substantial similarities were seen between rural and urban CCIs, which suggests the need to broaden the discussion on CCIs to include research on rural initiatives in the CCI literature. PMID- 15537178 TI - A study of community guides: lessons for professionals practicing with and in communities. AB - A study of 35 nonprofessional helpers, identified as community "guides," focused on the contribution each made to helping marginalized individuals and families become a part of their communities. The lessons learned through these lay helpers can inform a postmodern social work practice that promotes the use of indigenous practice principles appropriate for work with and in culturally distinct communities. The practice wisdom of these guides demonstrates a need for professionals to reposition themselves in the associational life of a community, and to make their practice less visible. It is shown that an effective community building practice that respects community solutions to individual and community problems requires permeable boundaries on the part of intervening professionals. PMID- 15537179 TI - Privatized management in urban public housing: a comparative analysis of social service availability, utilization, and satisfaction. AB - This article presents the results of a study that assessed the effect of privatized management on social service availability, utilization, and resident satisfaction in public housing communities. The respondents were heads of household who lived in public housing "projects" in Miami, Florida-more than 90 percent of whom were African American women. A quasi-experimental design with nonequivalent control groups compared respondent data from privately and publicly managed sites. Publicly managed sites reported more availability of social services, but privately managed sites reported higher levels of use. Implications for welfare reform, public housing demolition or renewal policies, the importance of social services with self-sufficiency efforts, and the need for culturally sensitive social work practices in ethnic minority public housing communities are discussed. PMID- 15537180 TI - A user-friendly approach to program evaluation and effective community interventions for families at risk of homelessness. AB - This article demonstrates how a user-friendly evaluation of a federally funded homeless prevention program using an action research approach--and using a logic model as the analytic framework--informed multiple stakeholders, including members of Congress, other decision makers, and Family Center practitioners. The program's target population was very low-income families at risk of being evicted from public housing. The authors discuss the methods used, the application of the logic model, and the study's findings as they unfolded in four phases: (1) logic modeling as program planning, (2) conceptualizing the intervention, (3) delineating implementation processes, and (4) determining the range of client outcomes. Implications for social policy, social work practice, and evaluation research are discussed. PMID- 15537181 TI - The Kosovo family support project: offering psychosocial support for families with missing persons. AB - This article presents a case that illustrates the vital role that social workers and other mental health and public health workers provided to support families following the 1999 conflict between Serb forces and Kosovar Albanians in Kosovo. A public health and social work team from Boston-based Physicians for Human Rights responded to an emergency postconflict situation by planning and implementing a community-based intervention that mobilized local mental health providers and established collaboration among national and international agencies, including the International War Crimes Tribunal for the Former Yugoslavia, to provide a support network for families with missing family members. The intervention included assessment of local resources, training, fieldwork, professional development of local staff, and the establishment of long term local responsibility and autonomy. PMID- 15537182 TI - Guidelines for ethical practice in community organization. AB - Community organizers often encounter ethical dilemmas in practice. Most organizers engage on a regular basis with community residents, constituency groups, local institutions, and government decision makers. Consequently, most practice activities occur outside traditional agency settings and are not directly addressed in the Code of Ethics of the National Association of Social Workers. Although community practice principles such as self-determination, informed consent, and protection of confidentiality are identified in the Code, situational factors make their application different than in direct practice. This article identifies the values inherent in community practice, describes ethical issues encountered by organizers, and examines tools available to organizers for resolving common ethical dilemmas. PMID- 15537183 TI - Innovations in the management of child protection workers: building worker resilience. AB - CPS staff work in a fishbowl. Parents, teachers, advocates, counselors, doctors, and lawyers frequently see themselves as experts in the field of CPS. Yet, few of them have had the experience of a CPS worker telling them how to do their job. The true potential for CPS work will not be realized until our communities decide to increase worker pay and reduce worker caseloads. Yet, if we choose to work under these conditions, it is our responsibility to find ways to survive them. The CPS workers unanimously reported that the overall impact of these progressive management strategies was to significantly reduce their stress. Given the serious threats, criticisms, and outrage thrown daily at CPS workers, it is imperative that managers act first to protect their worker's "best interests." It is my belief that all of the policy deviations described above are ethical. None of them put children at risk. I hope that policymakers will someday change the old school, management policies that are pervasive within our bureaucracies. I do not promote a wanton disregard for policy compliance; however, I do promote change where change is due. In light of the enormous impact that CPS workers have on children and their families, it is crucial that CPS managers be allowed to create an atmosphere where workers can successfully do their work. PMID- 15537184 TI - Social work misconduct may lead to liability. PMID- 15537185 TI - Stereotactic radiation treatment planning and follow-up studies involving fused multimodality imaging. AB - OBJECT: Innovative new software solutions may enable image fusion to produce the desired data superposition for precise target definition and follow-up studies in radiosurgery/stereotactic radiotherapy in patients with intracranial lesions. The aim is to integrate the anatomical and functional information completely into the radiation treatment planning and to achieve an exact comparison for follow-up examinations. Special conditions and advantages of BrainLAB's fully automatic image fusion system are evaluated and described for this purpose. METHODS: In 458 patients, the radiation treatment planning and some follow-up studies were performed using an automatic image fusion technique involving the use of different imaging modalities. Each fusion was visually checked and corrected as necessary. The computerized tomography (CT) scans for radiation treatment planning (slice thickness 1.25 mm), as well as stereotactic angiography for arteriovenous malformations, were acquired using head fixation with stereotactic arc or, in the case of stereotactic radiotherapy, with a relocatable stereotactic mask. Different magnetic resonance (MR) imaging sequences (T1, T2, and fluid attenuated inversion-recovery images) and positron emission tomography (PET) scans were obtained without head fixation. Fusion results and the effects on radiation treatment planning and follow-up studies were analyzed. The precision level of the results of the automatic fusion depended primarily on the image quality, especially the slice thickness and the field homogeneity when using MR images, as well as on patient movement during data acquisition. Fully automated image fusion of different MR, CT, and PET studies was performed for each patient. Only in a few cases was it necessary to correct the fusion manually after visual evaluation. These corrections were minor and did not materially affect treatment planning. High-quality fusion of thin slices of a region of interest with a complete head data set could be performed easily. The target volume for radiation treatment planning could be accurately delineated using multimodal information provided by CT, MR, angiography, and PET studies. The fusion of follow-up image data sets yielded results that could be successfully compared and quantitatively evaluated. CONCLUSIONS: Depending on the quality of the originally acquired image, automated image fusion can be a very valuable tool, allowing for fast (approximately 1-2 minute) and precise fusion of all relevant data sets. Fused multimodality imaging improves the target volume definition for radiation treatment planning. High-quality follow-up image data sets should be acquired for image fusion to provide exactly comparable slices and volumetric results that will contribute to quality contol. PMID- 15537186 TI - Significance of different conformity indices for evaluation of radiosurgery treatment plans for vestibular schwannomas. AB - OBJECT: There are various kinds of conformity parameters currently in use, although several of them are limited and reflect only target volume coverage or normal tissue overdosage. Indices are reviewed with the goal of determining those that are most significant for the evaluation of radiosurgery treatment plans for patients with vestibular schwannoma, based on the authors' experience at the Novalis Shaped Beam Surgery Center. METHODS: Fifty-five radiosurgery plans for patients with vestibular schwannomas (VSs) have been evaluated. In this paper the conformation number (CN) and dose-related CN (dCN) are evaluated, and a penalty for underdosed target volumes and overdosed normal tissue is incorporated. A strategy is discussed to apply these indices (CN and dCN) to define the optimal prescription isodose (PI). For a given radiosurgery treatment plan, permitting partial target underdosage may offer an improvement of the CN. Variations of different conformation indices have been calculated for varying prescription levels--for example, an isodose plan. The resulting graph for the CN is discussed in detail to illustrate its use in defining the optimal PI level. For the 55 cases of VSs reported on, the median CNmax result was 0.78. CONCLUSIONS: It is possible to achieve highly conformal dose distributions with Novalis radiosurgical system. The CN is the parameter of choice when evaluating radiosurgery treatment plans and scoring possible treatment plans. It takes into account both target underdosage and normal tissue overdosage and offers a valuable scoring parameter while avoiding false-perfect scores. PMID- 15537187 TI - First experience in using Novalis shaped beam radiosurgery in Korea. AB - OBJECT: The role of radiosurgery has become increasingly important in the treatment of intracranial lesions. In this study the authors evaluated the efficacy of the Novalis shaped beam radiosurgery system (dedicated linear accelerator) for various brain and spinal lesions. METHODS: Between November 2000 and October 2003 the authors treated 356 cases of various intracranial and extracranial lesions with Novalis shaped beam radiosurgery. Of these 356 cases, 109 cases were followed for more than a 2-year period. Fifty patients underwent fractionated stereotactic radiosurgery. Twenty patients (40%) harbored benign tumors (two acoustic neuromas, seven meningiomas, five pituitary adenomas, four optic gliomas, and two craniopharyngiomas), 18 patients (36%) harbored malignant tumors (16 glioblastoma multiforme [GBM], and two metastases), and the others included five with brainstem gliomas, one chordoma, five gliomas, and one with an arteriovenous malformation (AVM). Eighteen of 20 patients with benign tumors had good tumor control. Of 59 patients treated with single-dose stereotactic radiosurgery, 24 had benign intracranial tumors (12 acoustic neuromas, 11 meningiomas, and one pituitary adenoma), 20 had malignant tumors (two GBMs and 18 metastases), and the others were eight AVMs, two glomus jugulare tumors, three lymphomas, one pineal tumor, and one spinal tumor. CONCLUSIONS: Stereotactic radiosurgery and fractionated stereotactic radiosurgery using the Novalis shaped beam radiosurgery system are effective and safe noninvasive treatment modalities for various intracranial and extracranial lesions. PMID- 15537188 TI - Treatment of trigeminal neuralgia with linear accelerator radiosurgery: initial results. AB - OBJECT: Radiosurgery has emerged as an important treatment of trigeminal neuralgia. Substantial advantages have been demonstrated in safety and comfort over other modalities. Radiosurgical treatment of trigeminal neuralgia has been well investigated with gamma knife devices involving fixed cobalt sources. Few reports exist concerning trigeminal neuralgia treated using linear accelerator (LINAC)-based devices. In recent years these devices have reached the level of mechanical precision that is required for such functional treatments. The authors describe their initial experience with radiosurgical treatment of trigeminal neuralgia when using a BrainLAB Novalis LINAC device equipped with the commercially available 4-mm collimator. METHODS: A total of 32 patients were treated in a 12-month period between November 2002 and November 2003. The median patient age was 67 years (range 38-84 years). Facial pain was graded using the Barrow Neurological Institute (BNI) scoring system. All patients' pain was BNI Grade IV or V prior to treatment. Of these patients, 22 were undergoing initial treatment, and 10 were undergoing retreatment for recurrent pain following various treatments including percutaneous procedures, gamma knife surgery (GKS), or microvascular decompression. Two patients had multiple sclerosis. In patients undergoing initial radiosurgery, the most proximal segment of the cisternal portion of the trigeminal nerve received 85 to 90 Gy administered in a 5- or 7 noncoplanar arc single-isocenter plan with a 4-mm circular collimator. In patients undergoing repeated radiosurgery, the target received 60 Gy. Overall good and excellent results (BNI Grade I, II, or III) were achieved in 25 (78%) of 32 patients. The median time to pain relief was 6 weeks. Fair results (improvement in pain with BNI Grade IV) were achieved in three patients (9%), and poor results (no improvement in pain and BNI Grade IV or V) were seen in four (13%). Two patients demonstrated new trigeminal dysfunction following treatment. No other complications occurred. CONCLUSIONS: High-precision imaging and LINAC instrumentation have allowed for treatment of trigeminal neuralgia with results and safety comparable to those achieved using GKS. Linear accelerator-based radiosurgery with the Novalis device is a safe and effective method of managing trigeminal neuralgia and may become the preferred means at centers where the technology is available. PMID- 15537189 TI - Geometrical accuracy of the Novalis stereotactic radiosurgery system for trigeminal neuralgia. AB - OBJECT: Stringent geometrical accuracy and precision are required in the stereotactic radiosurgical treatment of patients. Accurate targeting is especially important when treating a patient in a single fraction of a very high radiation dose (90 Gy) to a small target such as that used in the treatment of trigeminal neuralgia (3 to 4-mm diameter). The purpose of this study was to determine the inaccuracies in each step of the procedure including imaging, fusion, treatment planning, and finally the treatment. The authors implemented a detailed quality-assurance program. METHODS: Overall geometrical accuracy of the Novalis stereotactic system was evaluated using a Radionics Geometric Phantom Chamber. The phantom has several magnetic resonance (MR) and computerized tomography (CT) imaging-friendly objects of various shapes and sizes. Axial 1-mm thick MR and CT images of the phantom were acquired using a T1-weighted three dimensional spoiled gradient recalled pulse sequence and the CT scanning protocols used clinically in patients. The absolute errors due to MR image distortion, CT scan resolution, and the image fusion inaccuracies were measured knowing the exact physical dimensions of the objects in the phantom. The isocentric accuracy of the Novalis gantry and the patient support system was measured using the Winston-Lutz test. Because inaccuracies are cumulative, to calculate the system's overall spatial accuracy, the root mean square (RMS) of all the errors was calculated. To validate the accuracy of the technique, a 1.5 mm-diameter spherical marker taped on top of a radiochromic film was fixed parallel to the x-z plane of the stereotactic coordinate system inside the phantom. The marker was defined as a target on the CT images, and seven noncoplanar circular arcs were used to treat the target on the film. The calculated system RMS value was then correlated with the position of the target and the highest density on the radiochromic film. The mean spatial errors due to image fusion and MR imaging were 0.41+/-0.3 and 0.22+/-0.1 mm, respectively. Gantry and couch isocentricities were 0.3+/-0.1 and 0.6+/-0.15 mm, respectively. The system overall RMS values were 0.9 and 0.6 mm with and without the couch errors included, respectively (isocenter variations due to couch rotation are microadjusted between couch positions). The positional verification of the marker was within 0.7+/-0.1 mm of the highest optical density on the radiochromic film, correlating well with the system's overall RMS value. The overall mean system deviation was 0.32+/-0.42 mm. CONCLUSIONS: The highest spatial errors were caused by image fusion and gantry rotation. A comprehensive quality-assurance program was developed for the authors' stereotactic radiosurgery program that includes medical imaging, linear accelerator mechanical isocentricity, and treatment delivery. For a successful treatment of trigeminal neuralgia with a 4-mm cone, the overall RMS value of equal to or less than 1 mm must be guaranteed. PMID- 15537190 TI - A methodology for verification of radiotherapy dose calculation. AB - OBJECT: A methodology for dosimetric verification of radiation therapy plans was developed and implemented. Dosimetric accuracy of clinically active intensity modulated radiotherapy (IMRT) and intensity-modulated radiosurgery (IMRS) programs was assessed using this methodology. METHODS: The methodology included several dosimetric tasks that were performed to assess the dosimetric accuracy of a treatment plan. Absolute dosimetry of the composite plan was performed using an ionization chamber. Film dosimetry was performed for each individual field and for the multifield composite plan. Calculated dose distributions and film measurements were compared using software developed for the specific tasks. Two dimensional maps of gamma index, dose difference, and distance-to-agreement were calculated and displayed. To date, good agreement between measurements and calculations has been observed in 160 clinical IMRT and IMRS plans. The largest observed absolute dose disagreement was -4.79%. The mean absolute dose difference was 0.26%, with a standard deviation of 1.75%. The authors specify a 3% dose difference and 3-mm distance as the scaling acceptability criteria for the gamma index calculations of the film measurement analysis. The planning and delivery system in clinical use has proven consistently to satisfy these criteria. CONCLUSIONS: The dosimetric verification methods and the software tools developed were both quantitative and clinically practical. The measurements and the analysis demonstrated that the IMRT and IMRS planning and delivery system in use was sufficiently accurate for highly conformal treatments. PMID- 15537191 TI - Stereotactic radiotherapy for the treatment of acoustic neuromas. AB - OBJECT: The authors sought to assess the safety and efficacy of stereotactic radiotherapy when using a linear accelerator equipped with a micromultileaf collimator for the treatment of patients with acoustic neuromas. METHODS: Fifty patients harboring acoustic neuromas were treated with stereotactic radiotherapy between September 1997 and June 2003. Two patients were lost to follow-up review. Patient age ranged from 20 to 76 years (median 59 years), and none had neurofibromatosis. Forty-two patients had useful hearing prior to stereotactic radiotherapy. The fifth and seventh cranial nerve functions were normal in 44 and 46 patients, respectively. Tumor volume ranged from 0.3 to 19.25 ml (median 2.51 ml). The largest tumor dimension varied from 0.6 to 4 cm (median 2.2 cm). Treatment planning in all patients included computerized tomography and magnetic resonance image fusion and beam shaping by using a micromultileaf collimator. The planning target volume included the contrast-enhancing tumor mass and a margin of normal tissue varying from 1 to 3 mm (median 2 mm). All tumors were treated with 6-MV photons and received 54 Gy prescribed at the 90% isodose line encompassing the planning target volume. A sustained increase greater than 2 mm in any tumor dimension was defined as local relapse. The follow-up duration varied from 6 to 74 months (median 36 months). The local tumor control rate in the 48 patients available for follow up was 100%. Central tumor hypodensity occurred in 32 patients (67%) at a median of 6 months following stereotactic radiotherapy. In 12 patients (25%), tumor size increased 1 to 2 mm at a median of 6 months following stereotactic radiotherapy. Increased tumor size in six of these patients was transient. In 13 patients (27%), tumor size decreased 1 to 14 mm at a median of 6 months after treatment. Useful hearing was preserved in 39 patients (93%). New facial numbness occurred in one patient (2.2%) with normal fifth cranial nerve function prior to stereotactic radiotherapy. New facial palsy occurred in one patient (2.1 %) with normal seventh cranial nerve function prior to treatment. No patient's pretreatment dysfunction of the fifth or seventh cranial nerve worsened after stereotactic radiotherapy. Tinnitus improved in six patients and worsened in two. CONCLUSIONS: Stereotactic radiotherapy using field shaping for the treatment of acoustic neuromas achieves high rates of tumor control and preservation of useful hearing. The technique produces low rates of damage to the fifth and seventh cranial nerves. Long-term follow-up studies are necessary to confirm these findings. PMID- 15537192 TI - Functional stereotactic radiosurgery involving a dedicated linear accelerator and gamma unit: a comparison study. AB - OBJECT: The purpose of this work was to investigate the targeting and dosimetric characteristics of a linear accelerator (LINAC) system dedicated for stereotactic radiosurgery compared with those of a commercial gamma knife (GK) unit. METHODS: A phantom was rigidly affixed within a Leksell stereotactic frame and axial computerized tomography scans were obtained using an appropriate stereotactic localization device. Treatment plans were performed, film was inserted into a recessed area, and the phantom was positioned and treated according to each treatment plan. In the case of the LINAC system, four 140 degrees arcs, spanning +/-60 degrees of couch rotation, were used. In the case of the GK unit, all 201 sources were left unplugged. Radiation was delivered using 3- and 8-mm LINAC collimators and 4- and 8-mm collimators of the GK unit. Targeting ability was investigated independently on the dedicated LINAC by using a primate model. Measured 50% spot widths for multisource, single-shot radiation exceeded nominal values in all cases by 38 to 70% for the GK unit and 11 to 33% for the LINAC system. Measured offsets were indicative of submillimeter targeting precision on both devices. In primate studies, the appearance of an magnetic resonance imaging enhancing lesion coincided with the intended target. CONCLUSIONS: Radiosurgery performed using the 3-mm collimator of the dedicated LINAC exhibited characteristics that compared favorably with those of a dedicated GK unit. Overall targeting accuracy in the submillimeter range can be achieved, and dose distributions with sharp falloff can be expected for both devices. PMID- 15537193 TI - A feasibility study of 18F-fluorodeoxyglucose positron emission tomography targeting and simultaneous integrated boost for intensity-modulated radiosurgery and radiotherapy. AB - OBJECT: The authors hypothesized that the efficacy of intensity-modulated radiation therapy (IMRT) can be enhanced by selectively increasing the radiation dose to the biologically active positron emission tomography (PET)-documented positive tumor subregions while simultaneously maintaining the overall clinically established target dose. METHODS: The authors undertook a feasibility study to evaluate IMRT PET/computerized tomography (CT) protocol for boost treatment in selected cancer patients. Prior to treatment, FDG-PET and CT scans were acquired using an integrated PET/CT scanner, ensuring accurate correlation between image sets. After acquisition, tumor volume and objects-at-risk (OARs) were outlined on the CT scans; any PET-positive tumor subregions were similarly outlined. Daily dosages of 1.8 to 2 Gy were prescribed to tumor volume and the margin whereas additional dosages of 10 to 20% were delivered to PET-positive subregions. Dosage volume histogram-derived constraints were used in inverse planning to specify the desired dose to one or more PET-positive tumor subregions, CT-delineated tumor volume, and OARs. The IMRT treatment was delivered using a micromultileaf collimator. Simultaneous integrated boost radiation was successfully delivered using IMRT with PET/CT planning. Excellent dose conformality was achieved in the tumor volume and the dose to PET-positive tumor subregions was increased while minimizing the dose to OARs. CONCLUSIONS: When coupled with IMRT, PET/CT scanning allows dose escalation to biologically active subregions within the tumor volume. Further study is needed to determine if dose escalation to FDG-PET-active sites correlates with improved treatment outcome. Finally, in extracranial sites, PET scanning should only be performed with a dedicated PET/CT device because present image fusion technologies are inadequate for accurately registering deformable objects. PMID- 15537194 TI - Dose fractionation in stereotactic radiotherapy for parasellar meningiomas: radiobiological considerations of efficacy and optic nerve tolerance. AB - OBJECT: Benign meningiomas have been shown to be equally well controlled with single-dose radiosurgery (15 Gy) and fractionated doses of 54 Gy in 30 fractions after adequate follow up. For a subset of patients with meningioma, the optic apparatus is dose limiting when considering single-dose stereotactic radiosurgery, with tolerance estimated to be 8 to 10 Gy. Recently, hypofractionated regimens have been used to treat benign meningiomas with a small number of fractions. An analysis of the expected efficacy of hypofractionation compared with the estimated optic tolerance to fractionated radiotherapy was undertaken. METHODS: Using the assumption that 15 Gy in one fraction and 54 Gy in 30 fractions are isoeffective for control of benign meningioma, an alpha/beta for meningioma is calculated to be 3.28 Gy. Invoking a 10% error for these doses (15 Gy+/-10% is equivalent to 54 Gy+/-10%) results in upper and lower limits of the estimate for alpha/beta of 3.85 Gy and 2.7 Gy. Using these estimates, isoeffect curves for control of meningioma were constructed for fraction numbers of one to 45. Best estimates of optic nerve/chiasm tolerance to single doses of radiation are 8 to 10 Gy, with the reported incidence of optic neuropathy increasing significantly at higher doses. This is consistent with the optic ret model, which also predicts for optic tolerance following fractionated radiotherapy. Comparison of optic tolerance and estimates of efficacious doses at fraction numbers between one and 30 were made. Statistical estimates of patient numbers and duration of follow up required to rule out optic neuropathy following radiotherapy were made. Single doses of radiation required to treat benign meningioma optimally (13.5 16.5 Gy) clearly exceed the estimated and reported clinical tolerance of the optic nerves and chiasm. The application of equivalent biological doses in a small number of fractions continues to exceed optic tolerance until at least 25 fractions are applied. CONCLUSIONS: The use of small numbers of fractions to treat patients with meningioma when portions of optic nerve or chiasm receive full dose may result in undertreatment of the tumor and/or exceeding optic nerve tolerance. In such cases standard fractionation is recommended. Ruling out a low, yet unacceptable, risk of optic neuropathy may require the close study of many patients with long-term follow-up evaluation. PMID- 15537195 TI - Use of stereotactic intensity-modulated radiotherapy in thyroid-related ophthalmopathy. Case report. AB - Thyroid-related ophthalmopathy (TRO), a debilitating condition involving a range of visual and orbital symptoms, occurs in up to 40% of patients with Graves disease. The goals of treatment include correcting thyroid dysfunction, relieving ocular pain, preserving vision, and improving cosmetic appearance. Options for therapy include symptomatic treatment, glucocorticoid medication, radiation therapy, and surgery. Traditional radiation treatment uses small opposed bilateral fields consisting of retrobulbar volumes and customized blocks to shield periorbital structures. The combination of intensity-modulated radiotherapy (IMRT) and stereotactic technology facilitates the administration of radiation to patients suffering from TRO and provides greater safety and efficacy than traditional treatment. The authors present the case of a patient with severe TRO whose symptoms resolved rapidly after treatment with stereotactic IMRT. The outcome in this case supports stereotactic IMRT as an effective treatment option for patients with TRO who also undergo radiation therapy. PMID- 15537196 TI - Patterns of failure after single-dose radiosurgery for spinal metastasis. AB - OBJECT: Single-dose radiosurgery for solitary spinal metastases can achieve rapid and durable pain control. This study was conducted to determine the patterns of failure after spinal radiosurgery. METHODS: Forty-nine patients with 61 solitary spinal metastases underwent radiosurgery between May 2001 and May 2003. Single dose radiosurgery (10-16 Gy) was delivered only to the involved spinal segments. The authors undertook a retrospective review of clinical notes, including patient questionnaires and radiological studies (computerized tomography or magnetic resonance imaging), to analyze patterns of failure following radiosurgery with regard to the pain and tumor control. Complete and partial pain relief was achieved in 85% of the lesions treated. Relapse of pain at the treated site was noted in 7%. Radiologically, lesions progressively metastasized to the immediately adjacent spines in 5%. These patients also had progressive primary and/or other systemic metastatic diseases. CONCLUSIONS: Spine-related pain control/reduction was excellent. Tumor recurrence at the treated segment and progression to the immediately adjacent region were rare. The results support the use of spinal radiosurgery as an effective treatment option for solitary spinal metastasis. PMID- 15537197 TI - Long-term management of patients with multiple brain metastases after shaped beam radiosurgery. Case report and review of the literature. AB - The role of radiosurgery in the treatment of patients with advanced-stage metastatic disease is currently under debate. Previous randomized studies have not consistently supported the use of radiosurgery to treat patients with numbers of brain metastases. In negative-results studies, however, intracranial tumor control was high but extracranial disease progressed; thus, patient survival was not greatly affected, although neurocognitive function was generally maintained until death. Because the future promises improved systemic (extracranial) therapy, the successful control of brain disease is that much more crucial. Thus, for selected patients with multiple metastases to the brain who remain in good neurological condition, aggressive lesion-targeting radiosurgery should be very useful. Although a major limitation to success of this therapy is the lack of control of extracranial disease in most patients, it is clear that well-designed, aggressive treatment substantially decreases the progression of brain metastases and also improves neurocognitive survival. The authors present the management and a methodology for rational treatment of a patient with breast cancer who has harbored 24 brain metastases during a 3-year period. PMID- 15537198 TI - Safety and efficacy of stereotactic radiosurgery for tumors of the spine. AB - OBJECT: The extension of stereotactic radiosurgery treatment of tumors of the spine has the potential to benefit many patients. As in the early days of cranial stereotactic radiosurgery, however, dose-related efficacy and toxicity are not well understood. The authors report their initial experience with stereotactic radiosurgery of the spine with attention to dose, efficacy, and toxicity. METHODS: All patients who underwent stereotactic radiosurgery of the spine were treated using the Novalis unit at Westchester Medical Center between December 2001 and January 2004 are included in a database consisting of demographics on disease, dose, outcome, and complications. A total of 31 patients (12 men, 19 women; mean age 61 years, median age 63 years) received treatment for 35 tumors. Tumor types included 26 metastases (12 lung, nine breast, five other) and nine primary tumors (four intradural, five extradural). Thoracic tumors were most common (17 metastases and four primary) followed by lumbar tumors (four metastases and four primary). Lesions were treated to the 85 to 90% isodose line with spinal cord doses being less than 50%. The dose per fraction and total dose were selected on the basis of previous treatment (particularly radiation exposure), size of lesion, and proximity to critical structures. CONCLUSIONS: Rapid and significant pain relief was achieved after stereotactic radiosurgery in 32 of 34 treated tumors. In patients treated for metastases, pain was relieved within 72 hours and remained reduced 3 months later. Pain relief was achieved with a single dose as low as 500 cGy. Spinal cord isodoses were less than 50% in all patients except those with intradural tumors (mean single dose to spinal cord 268 cGy and mean total dose to spinal cord 689 cGy). Two patients experienced transient radiculitis (both with a biological equivalent dose (BED) > 60 Gy). One patient who suffered multiple recurrences of a conus ependymoma had permanent neurological deterioration after initial improvement. Pathological evaluation of this lesion at surgery revealed radiation necrosis with some residual/recurrent tumor. No patient experienced other organ toxicity. Stereotactic radiosurgery of the spine is safe at the doses used and provides effective pain relief. In this study, BEDs greater than 60 Gy were associated with an increased risk of radiculitis. PMID- 15537199 TI - Image-guided procedures for intensity-modulated spinal radiosurgery. Technical note. AB - Radiosurgery for brain tumors has been well established in the radiation oncology and neurosurgery fields. Radiosurgery of extracranial tumors such as those involving the spine is, however, still in the early stage because of difficulties in patient immobilization and organ motion. The authors describe an image-guided procedure for intensity-modulated spinal radiosurgery that was developed at Henry Ford Hospital. PMID- 15537200 TI - Novalis Shaped Beam Radiosurgery of arteriovenous malformations. AB - OBJECT: The authors studied outcomes and complications in patients who harbored arteriovenous malformations (AVMs) and underwent stereotactic radiosurgery involving the Novalis shaped beam unit. METHODS: Between January 1998 and January 2002, 83 patients were treated with radiosurgery at University of California, Los Angeles. The mean patient age was 37.8 years. Forty-four patients completed follow up. There were 24 women. Sixteen patients underwent repeated radiosurgery. Embolization was performed in 13 patients and radiosurgery alone in 31. The mean follow-up period after embolization was 54.4+/-21.9 months and 37.4+/-14.6 months for radiosurgery alone. The mean peripheral dose was 15 Gy (range 12-18 Gy). The mean preradiosurgery lesion volume was 9.7+/-11.9 ml for radiosurgery alone and 16.2+/-11.3 ml for embolization. The AVMs in 13 patients (29.8%) were Spetzler Martin Grade II, 12 (27.5%) were Grade III, eight (18.2%) Grade IV, and five (11.3%) were Grade V and VI each. Spetzler-Martin grade, volume, and peripheral dose were analyzed in consideration to outcome. A positive trend (p = 0.086) was observed between Spetzler-Martin grade and obliteration rate. Volume per se did not predict obliteration (p = 0.48). A peripheral dose of 18 Gy was shown to be the most important predictor for occlusion (p = 0.007). The overall obliteration rate was 52.5%. A transient complication was noticed in one case (2.3%) and but no permanent deficits due to radiosurgery have been detected so far. Three patients (6.8%) bled after radiosurgery. CONCLUSIONS: The range of the prescribed peripheral dose was narrow. An association between the mean peripheral dose of 15 Gy, high conformality, and homogeneous dose distribution permitted no permanent complications. Volume per se did not correlate with outcome. The next step will be to increase the peripheral dose shaping the beam and to achieve higher obliteration rates without increasing complications. PMID- 15537201 TI - Spinal lesions treated with Novalis shaped beam intensity-modulated radiosurgery and stereotactic radiotherapy. AB - OBJECT: Spinal radiosurgery was implemented to improve quality of life (QOL) in patients with malignancies. It may also be applicable to the treatment of benign lesions. METHODS: Between July 2002 and January 2004, 14 patients harboring 22 lesions were treated; 13 received single-dose stereotactic radiosurgery. Six were women. The mean age was 60.2 years (range 48-82 years). There were 11 metastases, two neurofibromas, and one meningioma. Six lesions were cervical, 10 thoracic, and six lumbar. Ten patients suffered pain, three paresthesias, two weakness, and three were asymptomatic. Seven patients underwent spinal surgery, with four receiving instrumentation. Twelve patients underwent conventional irradiation before stereotactic radiosurgery/stereotactic radiotherapy. A mean dose of 12+/ 2.7 Gy (range 8-21 Gy) was prescribed to the 91% isodose line (range 85-97%). The mean tumor volume was 25+/-27.1 ml (range 0.75-91.8 ml). Treatment was planned using intensity-modulated radiosurgery (IMRS) fields in 15 cases, dynamic arcs in five, and conformal beams in two. The mean follow-up period was 6.1+/-3.9 months (range 1-16 months). Three patients became pain free and four experienced considerable relief. Weakness improved in the two patients with this preoperative symptom and the asymptomatic patients remained so. Four lesions decreased in size, five remained stable, seven progressed, and six were not followed up (two patients died before follow up). Four patients in all died, three of systemic disease and one of thoracic lesion progression. No complications due to shaped beam and IMRS/intensity-modulated radiotherapy (IMRT) techniques were observed. CONCLUSIONS: Shaped beam and IMRS/IMRT involving the Novalis system may delay neurological deterioration, improving QOL. The lack of complication suggests that higher doses can be delivered to improve the control rate in patients with metastases. PMID- 15537202 TI - Communicating the logic of a treatment plan formulated in Asbru to domain experts. AB - This paper presents an interactive visualization for medical treatment plans that are formulated in the plan representation language Asbru. So far, most attention of the protocol-based care community was focused towards formal guideline representation and authoring partly supported by graphical tools. The intention of this work is to go the opposite way and communicate the logic of a computerized treatment plan to physicians, nursing-, and other medical personnel visually. The visualization is based on the idea of flow-chart algorithms widely used in medical education and practice. This concept has been extended in order to cope with the powerful and expressive guideline representation language Asbru. Furthermore, a number of interactive navigational and overview extensions are used to intuitively support the understanding of the logic of plans. The user centered development approach applied for these interactive visualization methods has been guided by user input gathered via a user study, design reviews, and prototype evaluations as described in this document. PMID- 15537203 TI - Decision support and knowledge management in oncology using hierarchical classification. AB - This paper presents the KASIMIR research project for the management of decision protocols in oncology. A decision protocol is a kind of decision tree implemented in an object-based representation formalism. A reasoner based on such a formalism and on hierarchical classification is coupled with a knowledge editor. This association provides an assistance for editing and maintenance of protocols, enabling the detection of errors and the comparison between versions of the protocol. In this way, a management of protocols takes fully advantage of the underlying knowledge representation and reasoning tools. This straightforward use of the protocol may be insufficient in some situations. Then, the protocol may have to be adapted for these situations. A study of protocol adaptation is presented. In particular a reasoner based on a combination of hierarchical classification and fuzzy logic is introduced. PMID- 15537204 TI - Modelling clinical goals: a corpus of examples and a tentative ontology. AB - Knowledge of clinical goals and the means to achieve them are either not represented in most current guideline representation systems or are encoded procedurally (e.g. as clinical algorithms, condition-action rules). There would be a number of major benefits if guideline enactment systems could reason explicitly about clinical objectives (e.g. whether a goal has been successfully achieved or not, whether it is consistent with prevailing conditions, or how the system should adapt to circumstances where a recommended action has failed to achieve the intended result). Our own guideline specification language, PROforma, includes a simple goal construct to address this need, but the interpretation is unsatisfactory in current enactment engines, and goals have yet to be included in the language semantics. This paper discusses some of the challenges involved in developing an explicit, declarative formalism for goals. As part of this, we report on a study we have undertaken which has identified over 200 goals in the routine management of breast cancer, and outline a tentative formal structure for this corpus. PMID- 15537205 TI - Standardized terminology for clinical trial protocols based on top-level ontological categories. AB - This paper describes a new method for the ontologically based standardization of concepts with regard to the quality assurance of clinical trial protocols. We developed a data dictionary for medical and trial-specific terms in which concepts and relations are defined context-dependently. The data dictionary is provided to different medical research networks by means of the software tool Onto-Builder via the internet. The data dictionary is based on domain-specific ontologies and the top-level ontology of GOL. The concepts and relations described in the data dictionary are represented in natural language, semi formally or formally according to their use. PMID- 15537206 TI - TimeWrap--a method for automatic transformation of structured guideline components into formal process-representations. AB - Guideline and protocol representation languages have reached a level of complexity where auxiliary methods are needed to support the authoring of protocols in the particular language. Several approaches and methods exist that claim high knowledge about both, the medical context and the formal requirements. Therefore, we need knowledge-based methods to facilitate the human plan designer and create the protocols of the particular language as automated as possible. We present a three-step wrapper method, called TimeWrap, to extract information, in particular temporal issues, out of semistructured data and integrate it in a formal representation. We illustrate our approach using the guideline representation language Asbru and examples from guidelines to treat conjunctivitis. PMID- 15537207 TI - Non-compliance with guidelines: motivations and consequences in a case study. AB - Guidelines are often based on a mixture of evidence-based and consensus-based recommendations. It is not straightforward that providing a series of "good" recommendations result in a guideline that is easily applicable, and it is not straightforward that acting according to such recommendations leads to an effective and efficient clinical practice. In this paper we summarize our experience in evaluating both the usability and the impact of a guideline for the acute/subacute stroke management. A computerised version of the guideline has been implemented and linked to the electronic patient record. We collected data on 386 patients. Our analysis highlighted a number of non-compliances. Some of them can be easily justified, while others depend only on physician resistance to behavioural changes and on cultural biases. From our results, health outcomes and costs are related to guideline compliance: a unit increase in the number of non compliance results in a 7% increase of mortality at six months. Patients treated according to guidelines showed a 13% increase in treatment effectiveness at discharge, and an average cost of 2929 Euros vs 3694 Euros for the others. PMID- 15537208 TI - Advanced temporal data abstraction for guideline execution. AB - Temporal data abstraction bridges the gap between snap shot values delivered by monitoring devices and laboratory tests on one side and high-level medical concepts used in guidelines and by medical professionals on the other side. Within this field, the detection and abstraction of repeated patterns is a complex and important challenge. A repeated pattern is a combination of events or intervals which occur multiple times in a formally describable temporal relation. While there are many approaches to detect patterns in time series without prior definition of target concepts, we describe the application of temporal data abstraction in the context of guideline execution. Here predefined concepts of temporal patterns must be compared with measurement series describing the patient state. We discuss the requirements coming from both high-frequency domains such as intensive care units and low-frequency domains such as diabetes monitoring and show our solution based on a new version of the Asgaard data abstraction unit. It interfaces the dynamically changing patient state to the guideline execution unit and features abstraction modules ranging from simple calculations to statistical measures calculated for sliding time windows. PMID- 15537209 TI - Protocure: supporting the development of medical protocols through formal methods. AB - Medical guidelines and protocols describe the optimal care for a specific group of patients and therefore, when properly applied, improve the quality of patient care. During the last decade, a large number of medical guidelines and protocols have been published. However, the work done on developing and disseminating them far outweighs the efforts on guaranteeing their quality. Indeed, anomalies like ambiguity and incompleteness are frequent in medical guidelines and protocols. An approach grounded on a formal representation, can answer these needs, as we have demonstrated in the Protocure project'. The Protocure II project will aim at integrating formal methods in the life cycle of guidelines. PMID- 15537210 TI - Towards a flexible integration of clinical guideline systems with medical ontologies and medical information systems. AB - The integration of a computer-based system dealing with clinical guidelines with a medical ontology can provide several advantages, including standardization and knowledge sharing. Furthermore, in order to operate in the clinical practice, guideline systems must also interact with the hospital databases to retrieve patients' data. Unfortunately, currently there seems not to be any "standard" consensus model either for the medical ontology or for (the conceptual structure of) patient databases (even if several interesting proposals have been carried out). In this paper we show how we are extending the GLARE guideline manager in order to strictly interact with both a medical ontology and a patient Database, in such a way that GLARE is not committed to any specific ontology and database (i.e., different ontologies and/or databases can be used). PMID- 15537211 TI - Adaptive guideline-based treatment workflows with AdaptFlow. AB - One goal in modern medicine is to increase the treatment quality. A major step towards this aim is to support the execution of standardized, guideline-based clinical protocols, which are used in many medical domains, e.g., for oncological chemotherapies. Standardized chemotherapy protocols contain detailed and structured therapy plans describing the single therapy steps (e.g., examinations or drug applications). Therefore, workflow management systems offer good support for these processes. However, the treatment of a particular patient often requires modifications due to unexpected infections, toxicities, or social factors. The modifications are described in the treatment protocol but not as part of the standard process. To be able to further execute the therapy workflows in case of exceptions running workflows have to be adapted dynamically. Furthermore, the physician should be supported by automated exception detection and decision support for derivation of necessary modifications. The AdaptFlow prototype offers the required support for the field of oncological chemotherapies by enhancing a workflow system with dynamic workflow adaptation and rule based decision support for exception detection and handling. PMID- 15537212 TI - A generic interface to XML documents for guidance information. AB - We present a generic means of interfacing XML documents and clinical systems. The interface has been developed to allow the integration of best practice guidance information within prescribing systems. The interface has the following characteristics: (1) integrating developers do not have to interpret the structure of the XML documents, (2) inconsistencies between integrations are reduced, (3) the structure of the XML documents can change without affecting integration and (4) specification and documentation of the NDR document interface is within the interface itself. PMID- 15537213 TI - Transforming written guidelines into electronic formats--international perspectives. AB - The Guidelines International Network (G-1-N www.g-i-n.net) is a major new international initiative involving guideline-developing organisations from around the world. G-I-N seeks to improve the quality of health care by promoting systematic development of clinical practice guidelines and their application into practice. The Network now has over 45 international members, most of whom prepare evidence-based clinical practice guidelines, or actively promote the use of evidence in practice. One of the priorities of the organisation is to share evidence tables and adapt guidelines for local circumstances based on international evidence. In the longer term, guideline developers are planning to create 'living guidelines' that can be continuously updated and used by a number of different countries. A major consideration for guideline developers is how to communicate and work with information technology scientists to develop standards and protocols for the translation of these trans-national guidelines into electronic formats. To be effective, there must be formal internationally agreed standards that allow electronic guidelines to be shared and automatically updated. The Guidelines International Network will be taking a leading international role in working with designers and vendors of electronic decision support systems and tools to guarantee the integrity of guidelines when translated into electronic formats. This presentation by Catherine Marshall, Kitty Rosenbrand and Guenter Ollenschlaeger will: --explore current experiences from New Zealand, Germany and the Netherlands --identify issues from the perspective of guideline developers --make recommendations for establishing opportunities for software designers, vendors and informatics experts to collaborate with guideline developers to ensure that up to date evidence can be easily implemented and shared throughout the world. PMID- 15537214 TI - A multiple-ontology customizable search interface for retrieval of clinical guidelines. AB - A major problem in the effective use of clinical guidelines is fast and accurate access at the point of care. Thus, we are developing a digital electronic guideline library (DeGeL) and a set of tools for incremental conversion of free text guidelines into increasingly machine-comprehensible representations, which support automated application. Even if guidelines are represented in electronic fashion, care providers need to be able to quickly retrieve the guidelines that best fit the clinical situation at hand. We describe Vaidurya, a search and retrieval engine that exploits the hybrid nature of guideline representation in the DeGeL architecture. Vaidurya can use not only free-text keywords, but also multiple semantic indices along which the guidelines are classified, and the mark up of guidelines in DeGeL, using the semantic roles of one or more guideline representation languages (ontologies). Vaidurya offers a wide variety of querying options, in order to enable different types of users to query the guideline library in a manner that is both efficient and user friendly. We describe the customizable query interface, in which each user can create their own personal query interface. PMID- 15537215 TI - Mark-up based analysis of narrative guidelines with the Stepper tool. AB - The Stepper tool was developed to assist a knowledge engineer in developing a computable version of narrative guidelines. The system is document-centric: it formalises the initial text in multiple user-definable steps corresponding to interactive XML transformations. In this paper, we report on experience obtained by applying the tool on a narrative guideline document addressing unstable angina pectoris. Possible role of the tool and associated methodology in developing a guideline-based application is also discussed. PMID- 15537216 TI - A description logics approach to CGPS. AB - We develop a formal framework by which clinical guidelines and protocols (CGPs) can be partially represented as a set of terminological concept definitions using standard description logics. There are two benefits in pursuing such an approach. First, it provides a foundation for logic-based CGP fusion and collision detection. Second, it allows for the checking of clinical treatment episodes from the EPR against CGPs. PMID- 15537217 TI - Reminder-based or on-demand decision support systems: a preliminary study in primary care with the management of hypertension. AB - ASTI is a guideline-based decision support system for therapeutic prescribing in primary care with two modes of interaction. The "critic mode" operates as a reminder system to detect non guideline-compliant physician drug orders, whereas the "guided mode" operates on demand and provides physician guidance to help her establishing best recommended drug prescriptions for the management of hypertension. A preliminary evaluation study was conducted with 10 GPs to test the complementary nature of both modes of decision support. Results tend to validate our assumption that reminder-based interaction is appropriate for simple cases and that physicians are willing to use on-demand systems as clinical situations become more complex. PMID- 15537218 TI - The Digital electronic Guideline Library (DeGeL): a hybrid framework for representation and use of clinical guidelines. AB - We propose to present a poster (and potentially also a demonstration of the implemented system) summarizing the current state of our work on a hybrid, multiple-format representation of clinical guidelines that facilitates conversion of guidelines from free text to a formal representation. We describe a distributed Web-based architecture (DeGeL) and a set of tools using the hybrid representation. The tools enable performing tasks such as guideline specification, semantic markup, search, retrieval, visualization, eligibility determination, runtime application and retrospective quality assessment. The representation includes four parallel formats: Free text (one or more original sources); semistructured text (labeled by the target guideline-ontology semantic labels); semiformal text (which includes some control specification); and a formal, machine-executable representation. The specification, indexing, search, retrieval, and browsing tools are essentially independent of the ontology chosen for guideline representation, but editing the semi-formal and formal formats requires ontology-specific tools, which we have developed in the case of the Asbru guideline-specification language. The four formats support increasingly sophisticated computational tasks. The hybrid guidelines are stored in a Web based library. All tools, such as for runtime guideline application or retrospective quality assessment, are designed to operate on all representations. We demonstrate the hybrid framework by providing examples from the semantic markup and search tools. PMID- 15537219 TI - Using a guideline-centered approach for the design of a clinical decision support system to promote smoking cessation. AB - Knowledge acquisition for the design of clinical decision support systems can be facilitated when clinical practice guidelines serve as a knowledge source. We describe application of the Guideline Elements Model (GEM) in the design of a decision support system to promote smoking cessation. Following selection of relevant recommendations and markup of knowledge components with the GEM Cutter editor, the Extractor stylesheet was used to create a list of decision variables and actions for further processing. Decision variables and actions that reflect similar concepts were consolidated. Action types were identified. Extracting the critical concepts from the narrative text facilitates clarification of necessary content. The guideline-centric approach promotes accurate translation of guideline knowledge. PMID- 15537220 TI - Analysis of guideline compliance--a data mining approach. AB - While guideline-based decision support is safety-critical and typically requires human interaction, offline analysis of guideline compliance can be performed to large extent automatically. We examine the possibility of automatic detection of potential non-compliance followed up with (statistical) association mining. Only frequent associations of non-compliance patterns with various patient data are submitted to medical expert for interpretation. The initial experiment was carried out in the domain of hypertension management. PMID- 15537221 TI - The GLARE approach to clinical guidelines: main features. AB - In this paper, we present GLARE, a domain-independent prototypical system for acquiring, representing and executing clinical guidelines. GLARE has been built within a 7-year project with Azienda Ospedaliera San Giovanni Battista in Turin (one of the largest hospitals in Italy) and has been successfully tested on clinical guidelines in different domains, including bladder cancer, reflux esophagitis, and heart failure. GLARE is characterized by the adoption of advanced Artificial Intelligence (AI) techniques, to support medical decision making and to manage temporal knowledge. PMID- 15537222 TI - The SAGE guideline modeling: motivation and methodology. AB - The SAGE (Standards-Based Sharable Active Guideline Environment) project is a collaboration among research groups at six institutions in the US. The ultimate goal of the project is to create an infrastructure that will allow execution of standards-based clinical practice guidelines across heterogeneous clinical information systems. This paper describes the design goals of the SAGE guideline model in the context of the technological infrastructure and guideline modeling methodology that the project is developing. PMID- 15537223 TI - Tracing the formalization steps of textual guidelines. AB - This paper presents a new guideline authoring tool, called Guideline Markup Tool (GMT). It proposes two useful features, which are missing in existing tools. First, it facilitates the translation of a free-text guideline into a formal representation, providing special XML macros. Second, it can be used to create links between the original guideline and its formal representation. Therefore, the GMT eases the implementation of clinical guidelines in a formal representation, which can be used in monitoring and therapy planning systems. PMID- 15537224 TI - Translating Arden MlMs into GLIF guidelines--a case study of hyperkalemia patient screening. AB - To re-examine the validity of the medical knowledge that are embedded in the legacy system, we translated a Medical Logic Module (MLM) for hyperkalemia patient screening into the GuideLine Interchange Format (GLIF). We used a set of guiding principles to direct the translation. In addition, we used the GLIF3 Guideline Execution Engine (GLEE) as a testing tool to validate the encoded GLIF guideline by applying it to 5 simulated patient cases. The result has shown that it is possible to translate Arden MLMs into GLIF guidelines. However, significant efforts are necessary to handle the problems arose during the translation process. Automatic translation could be a more generalizable approach for future work. PMID- 15537225 TI - A telemedicine guideline for the practice of teleconsultation. AB - To establish a guideline for the use of teleconsultation, which is one of the most important applications in telemedicine beeing perfomed very frequently in daily medical practice in almost any field of medicine. EVIDENCE: The recommendations are based on expert knowledge, because of the lack of evidence based data in the telemedical scientific literature. In addition, scientific articles of the highest level of evidence available published between 1970 and 1999 were reviewed. DEVELOPEMENT AND CONSENUS PROCESS: A guideline draft was prepared using the attributes of clinical practice guidelines developed by the Institute of Medicine (IOM) of the National Academy of Sciences. This draft was reviewed by telemedicine experts and the content of the guideline was approved by 100% group consensus in 5 meetings held between 1997 and 2000 by a panel of members of the Subproject 4 Group of the Global Health Care Application Project of the G8 countries. CONCLUSIONS: The guideline gives recommendations on all aspects of teleconsultation in any field of medicine and will be updated regularly implementing new evidence. PMID- 15537226 TI - Medical telematics in disaster response. AB - INTRODUCTION: Every year many disasters cause thousands of injuries, deaths, refugees. Depending on the kind of disaster (train/plane accident, flood, earthquake) not only an acute emergency medicine treatment but also general and family medicine and hospital treatment have to be safeguarded over a longer time period in the disaster area. PROBLEM: Regarding to a lot of organizations, institutions and disaster teams taking part in the disaster assistance is there any lack of work or data flow in the medical treatment? METHODS: From the ODRA flood 1997, the high speed train crash in ESCHEDE 1998, the DANUBE flood 1999 and the ELBE flood in 2002 experience reports were collected. They were analysed with emphasis on data and work flow in the medical treatment and its command system: Standardised command structure? Communication problems? Used communication lines? Language problems? Medical Intelligence distribution? Use of Patient Tracking System? Triage problems? RESULTS: The use of spoken radio communication causes transmission mistakes or misunderstandings and radio-overload and need connection set-up-time for each call. Manual distribution of same data for many receivers using different communication lines causes a time shift in the up-to-date information. Language problems during the ODRA flood between German and Polish people led to longer reaction times. Up-to-date triage results as well as up-to date transportation and hospital information are necessary for medical evacuation. Compared with other reports about these disasters the quality of disaster management depends on the quality of communication and information. CONCLUSION: The use of health telematics in disaster response helps to cope with the scenario. Modern technologies provide support for building up medical aid although the normal infrastructure is destroyed. To cope with disaster scenarios there are some telematic tools which can be used:--Computer-based Command and Control System--Telemedical support --Data-ressources-network /Medical Intelligence. A further study is recommended to evaluate the real impact of using these telematic tools in a disaster. PMID- 15537227 TI - Architecture and tools for open, interoperable and portable EHRs. AB - Electronic Health Record (EHR) systems provide the kernel application of health information systems and health networks which should be independent of complexity, localisation constraints, platforms, protocols, etc. Based on shared care information systems' requirements for high level interoperability, a generic component architecture has been introduced. For implementing, running and maintaining acceptable and useable health information systems components, all views of the ISO Reference Model--Open Distributed Processing have to be considered. Following the Model Driven Architecture paradigm, a reference model as well as concept-representing domain models both independent of platforms must be specified, which are combined and harmonised as well as automatically transferred into platform-specific models using appropriate tools. PMID- 15537228 TI - Telemedicine in extreme environments: analogs for space flight. AB - The integration of telecommunications and information systems into health care delivery in human space flight operations is not new. It has been an integral tool for over 45 years. During these past decades, numerous efforts have been conducted to further develop and promulgate telemedicine. The National Aeronautics and Space Administration (NASA) established a commercial space center in 1997, known as the Medical Informatics and Technology Applications Consortium (MITAC). MITAC has developed and conducted a variety of test beds in several international settings, including Russia, Ecuador and other extreme and remote environments. These test beds have been designed to evaluate and validate technologies and techniques that have application in the delivery and support of health care in unique environments. The characteristics of these test beds are analogous to what might be observed or experienced in low earth orbit or on space based platform. These include intermittent communications, low bandwidth, level of competency of the front line health worker, etc. These test beds have led to new approaches for the delivery of health care as well as enhanced education. These experiences have been beneficial in the promulgation of telemedicine as an effective tool and have provided new ideals for space exploration as well terrestrial medicine. This paper will highlight MITAC's test beds and their relationship to space exploration. PMID- 15537229 TI - Development of a robotic navigation and fracture fixation system. AB - The use of robotics in surgery is nothing new. However, there are areas of surgery, such as in fracture fixation, where robots have yet to be implemented. This paper considers the choice of robot, gripper and ancillary equipment together with navigation systems necessary for their application. Hitherto robots have seen operation in surgery only in cases where relatively low manipulation forces are required. Nothing yet exists with the capability of handling forces in excess of 200 Newton as would be required in the above scenario. Another encumbrance to robots which are already in medical use is the difficulty in programming. Unfortunately most of these robots are programmed by specialists for a particular application. However, there exists a number of robot programming languages, like Unimation VA-LII (recently superceded by Staubli V+), which do not require specialist knowledge. The application of industrial robots to the "heavier" side of modern surgery is without doubt technically realisable. The remainder of this research project aims to determine exactly which robots and what ancilliary equipment are needed and then to implement them, first on plastic models and later on cadavers. A second phase is expected to deal with type approval and a final third phase with operations on live patients. PMID- 15537230 TI - A readiness model for telehealth is it possible to pre-determine how prepared communities are to implement telehealth? AB - Telehealth "readiness" can be defined as the degree to which users, health care organizations, and the health system itself are prepared to participate and succeed in its application. This project developed a readiness model for rural/remote locations in Canada. Specifically defined groups or communities with shared characteristics within a rural geographical community (i.e. practitioners, patients, the public, and health care organizations) participated in key informant interviews, awareness sessions, focus groups, and face-to-face interviews. The data were examined and organized keeping in mind Weiss' Program's Theory of Change. This approach allowed concrete and abstract factors to be considered. The model that emerged suggests that there are four types of readiness for each of the defined communities: core, engagement, structural, and non-readiness. The "communities" share some readiness factors and risks, but also exhibit unique elements. This finding is critical to acknowledge when the goal is to implement a useful, effective, and sustainable telehealth system within remote settings. Study results hold a key to understanding why technology systems have failed in the past, in spite of dedicating considerable human and financial resources towards their implementation. Notations of these findings will be helpful in future telehealth implementations within rural and isolated areas. PMID- 15537231 TI - An automated diagnostic system for tubular carcinoma of the breast--an overview of approach and considerations. AB - A computer-based automated histopathology recognition system was developed to distinguish benign from malignant lesions. Tubular carcinoma of the breast, which has several reactive and neoplastic mimics, was selected as a model. Archival stained tumour sections from the United Kingdom National External Quality Assurance Scheme for breast pathology and supplementary material from external pathologists formed the study population. A diagnostic process similar to that employed by the histopathologist was adopted, viz, low-power feature extraction and analysis by cluster/glandular groupings followed by high-power confirmation. To circumvent problems of stain variability, greyscale quantisation of images was achieved through Karhunen-Loeve transformation with results suggesting that histological stains provide information primarily through contrast and not colour. Mean nearest neighbour and variance of cell nuclei distances were found to be 100% effective in distinguishing images which contained diffuse tumour, and no clustering. Gaussian smoothing followed by minimum variance quantisation allowed segmentation of gland clusters. Perona-Malik nonlinear diffusion filter employed prior to intensity thresholding and morphological filtering was 92% (7330/7973) effective in segmenting individual glands. In a set of 62 benign and 52 malignant gland clusters, the features found to discriminate tubular carcinoma from benign conditions included > 20% of glands with sharp-angled edge, cluster area > 150,000 pixels, ratio total gland area:total cluster area < 0.14, > 60 glands per cluster and the ratio average malignant gland area:benign gland area < 0.5. Suspicious clusters were subjected to high-power feature analysis for nuclear morphology, nucleoli detection and basement membrane assessment. Watershed thresholding achieved nuclear segmentation and nuclear area > 1.3x mean benign nuclear area was found to have a malignant likelihood ratio of 14.5. Progressive thresholding was used to detect nucleoli. Basement membrane was accentuated by colour segmentation and demonstrated 0.96 sensitivity, 0.89 specificity and 0.92 positive predictive value for distinguishing malignancy. PMID- 15537232 TI - Data analysis now and then: significant changes in approaches and results. AB - Modern data analysis is one of the many prerequisites for telemedical applications. Classical statistical methods alone are no longer sufficient to fulfill the various demands of modern analytical procedures. Cluster and association analysis among others have filled this gap and are capable of producing more adequate and better suitable results as well as to provide information not detectable in the past. PMID- 15537233 TI - Software agents in surgery: an update. AB - Intelligent and, thus, autonomously reacting software programs are capable of handling a lot of different tasks as has been realized in economics and network administration. The same so-called software agents can be used for a variety of organizational tasks in medicine. Some software agents already manage an individual patient's health care record from documentation to ambulant or stationary admission, surgical planning, and many other tasks which currently consume more than half of a physician's daily working time. Hence, not only a large potential of time, but also of economical savings result to the physicians' new disposition. PMID- 15537234 TI - Technologies for haptic systems in telemedicine. AB - The ability to image the elastic properties of tissue is potentially useful in a variety of applications. The field of elastic imaging has grown in response to the potential use of such information in medical diagnosis. Real time ultrasound elastography represents a recent development in determining strain and elasticity distributions. Nevertheless, commonly used imaging techniques rely on the interpretation of two dimensional visual data displayed on a video screen. In reality however, physicians often prefer tactile exploration making the simultaneous portrayal of both video and haptic information most desirable. Since the 1970's many alphanumeric to tactile data conversion methods have been investigated, mainly with the ultimate aim of assisting the blind. More recently, interest has been directed toward the display of pictures on haptically explorable surfaces--Tactile imaging. Such a system would allow surgeons to examine hard sectors contained within soft tissue, and thereby assist in operations held remotely. The expansion of ultrasound elastography to 3D formats would mean the ability to haptically explore regions of the body normally inaccessible to human hands. For three-dimensional imaging the acquisition of sequential tomographic slices using Elastography, combined with image segmentation, enables the reconstruction, quantification and visualisation of tumour volumes. In a collaborative project between four research institutes, the aim is to produce a prototype three dimensional tactile displays comprising electrically switchable micromachined cells, whose mechanical moduli are governed by phase changes experienced by electrorheological and/or magnetorheological fluids. This will be integrated with a sensory ultrasonic elastography in order to present the human fingers with controllable surfaces capable of emulating biological tissue, muscle and bone. PMID- 15537235 TI - Cybercare NDMs: an improved strategy for biodefense using information technologies. AB - The National Disaster Medical System (NDMS) was created in the early 1980's, and it was designed to meet the threats of the time. Today the threats are much less discreet and predictable. They are distributed; they move and spread quickly; and they walk silently among us. Specifically, biological agents are an enemy unlike any we have had to deal with before. They offer unique challenges that fly in the face of current doctrine. We must redesign the NDMS in order to contain and eliminate this new threat. Tools exist today capable of effectively coordinating distributed resources--even through containment borders. We need to strengthen our public health system, create a net-centric disaster management system, and blur the boundaries between local and federal resources. Ultimately we must move from an incremental, echelon-based response to an immediate, continuous response. This can be accomplished by adding inexpensive, well-established information technologies to the existing response system. PMID- 15537236 TI - A communication-theory based view on telemedical communication. AB - Communication theory based analysis sheds new light on the use of health telematics. This analysis of structures in electronic medical communication shows communicative structures with special features. Current and evolving telemedical applications are analyzed. The methodology of communicational theory (focusing on linguistic pragmatics) is used to compare it with its conventional counterpart. The semiotic model, the roles of partners, the respective message and their relation are discussed. Channels, sender, addressee, and other structural roles are analyzed for different types of electronic medical communication. The communicative processes are shown as mutual, rational action towards a common goal. The types of communication/texts are analyzed in general. Furthermore the basic communicative structures of medical education via internet are presented with their special features. The analysis shows that electronic medical communication has special features compared to everyday communication: A third participant role often is involved: the patient. Messages often are addressed to an unspecified partner or to an unspecified partner within a group. Addressing in this case is (at least partially) role-based. Communication and message often directly (rather than indirectly) influence actions of the participants. Communication often is heavily regulated including legal implications like liability, and more. The conclusion from the analysis is that the development of telemedical applications so far did not sufficiently take communicative structures into consideration. Based on these results recommendations for future developments of telemedical applications/services are given. PMID- 15537237 TI - Implementation of TeleCare services: benefit assessment and organisational models. AB - All industrial societies are ageing. This has profound socio-economic and health sector implications. Innovative services based on Information Society Technologies (IST), like telehomecare are regarded as promising avenues to follow both to allow (national) health systems to cope with these challenges and to improve the quality of life of chronically ill and frail older citizens. The aim of the TEN-HMS project is to convincingly prove that telemonitoring of congestive heart failure (CHF) patients at home can improve medical outcome for these patients as well as their quality of life and the efficiency of healthcare delivery processes. But this will not (yet) be enough for the sustained success of such a service. Unless it takes into account the interests of the various players in the health care arena and a long-term Business Case can be proven, it will be very difficult to integrate such services into routine health care delivery processes. Before developing concrete delivery models for such a telemonitoring service, the "players" directly involved in such a service need to be identified--customers/patients, health services providers, IT services suppliers, and public/private insurance funds as payers--and their assessment perspectives considered. Then four concrete telemonitoring delivery models and their probability of success are discussed. Our analysis suggests that telemonitoring will presently only be successful if the service delivery model applied reflects national health system idiosyncrasies, takes into account established organisational boundaries and adapts to patient quality of life and health professional preferences. In the longer term, the new paradigm of seamless, patient-centred care will, however, require new, more efficient service delivery models integrating all aspects of the health services value chain. PMID- 15537238 TI - LifeGuard--recording, evaluation and wireless transmission of medical data. AB - Early diagnosis is imperative in the quest for cures for or prevention of diseases. Medical doctors nowadays can measure a multiplicity of physical determinants such as blood pressure, insulin levels, etc. The sooner this is done the sooner therapy can begin and the sooner therapy begins the greater the chances of success, no matter what the disease. That is why self-sufficient home monitoring and emergency aid systems are becoming ever more important in medical health care. PMID- 15537239 TI - NOAH--a mobile emergency care system. AB - The German emergency care system is a very sophisticated one. However, negative headlines like "Emergency Patient Tourism" appearing from time to time, have provoked a thorough deficit analysis which revealed two weak points: communication and documentation. The communication system presently used is a rather outdated one employing analogue voice radio between the ambulance cars/helicopters and the dispatch center and telephone communication between the dispatch center and the emergency rooms. To document the emergency case the on scene physician is required to fill out a form. A survey showed that many of these forms are filled out incompletely and/or inconsistently or are even missing completely. NOAH, which means "Emergency Organization and Administration Aid" ("Notfall Organisations- und Arbeits-Hilfe" in German) intends to address both of these communication and documentation deficits. The on-scene physician is equipped with a mobile ruggedised PC with an internal digital radio-modern. It provides a direct, digital communication channel from the on-scene physician to the emergency physician starting from the first minutes of the treatment of the emergency patient. It also provides an easy-to-fill-out variant of the paper form mentioned above with an on-line help function and visual aids. A typical course of events with the communication part of NOAH is as follows. The on-scene physician is alarmed via NOAH and can obtain details of the emergency during the approach. He can enter the status codes ("on the move","arrived at the emergency scene","arrived at the patient", etc.) with NOAH. During the first minutes of the treatment of the patient the physician enters a so called "First Message", which requires only 10 to 15 seconds. This message contains basic information like sex, age and the injuries of the patient. It helps the dispatch center (if the on scene physician desires so) to make an informed recommendation where to bring the patient. This message is also forwarded to the emergency room of the destination hospital, where it can help to start appropriate preparations for the patient. When the on-scene physician has enough time, he can already bring up the documentation masks and give further information to the emergency room. PMID- 15537240 TI - [Low birth weight--can we manage this problem?]. AB - Lowering the rate o low birth weight including prematurity is the main aim of preconception and prenatal care. In 2001 in comparison to 2000 the rate of reducing of perinatal mortality in Poland decreased. Unexpectedly the low birth weight rate increased. It is alarming finding. The preparation and implementation of locally modified prophylactic programmes is strongly needed. PMID- 15537241 TI - [Developmental sequel of prematurity]. AB - Severe developmental impairment in children born as extremly prematures include cerebral palsy, subnormal cognitive function, deafness and blindness. The rate of severe disabilities range from 5 to 30%. This article reviews what is known about early brain injury and its developmental sequel. PMID- 15537242 TI - [The problem of prematurity in Poland]. AB - In Poland an impressive decline in perinatal and neonatal mortality rate has been observed during the past decade. A further improvement will be hindered by such new phenomena like multiple gestation following assisted procreation and a higher risk of preterm birth arising from it as well as age increase among pregnant women. Prematurity rate is also more and more frequently influenced by preterm births induced due to threat to the life of the mother or the fetus. All these factors should be taken into consideration in preventive actions and health programs over the mother and the child. PMID- 15537243 TI - [Organizational possibilities of reducing the incidence of preterm deliveries]. AB - Preventing preterm deliveries and reducing their incidence are priority issues important to reduce infant mortality and morbidity. Organizational possibilities of reducing the incidence of preterm deliveries are associated mainly with identifying risk factors, primary prevention, intensifying secondary prevention and regionalization of perinatal care. PMID- 15537244 TI - [Causative hormonal prevention of premature labour]. AB - Labour is the most important caesura in the life of every human being, as it is the ultimate test of the child's fetal maturity to self dependent life, whose onset is connected with a violent change in the circulatory system and the commencement of breathing. Maturity assessment scale comprises such features as, among others: posture, angle forearm, pulling an elbow to the middle line of the body, distribution of lanugo, plantar creases, breast development. They can be objectified through their technical quantification the same way as we give the appropriate number of grams and centimeters when assessing the weight and length of the newborn. However, two equally mature fetuses call differ in mass by 800g, body length by 6 cm, and gestational age by 6 weeks. Relativity of pregnancy duration clearly demonstrates that only the quantitativization of maturity enabled its prenatal assessment on the basis of the rate of increase of the spatial parameters of the baby developing in the mother's womb. The date of the end of pregnancy is determined depending equally on the child's fetal maturity and the mother's readiness to birth, where the placenta plays an important role, as its existence coincides with the individual duration of every pregnancy. The use of industrially synthesized hormones means that those pharmaceuticals contain the whole mixture of side-products of the synthesis of the hormones. The biologically alien corticoid analogues like dexamethasone and betamethasone must, when exerting prolonged action, cause unwanted side effects. In contrast the ACTH depot administration leads to desired production and secretion of corticoids through the control of the whole body function of the adrenals. Such naturally stimulated endogenous steroid hormones are free from unwanted side effects of drugs from the group of their synthetic analogues. An enormous body of evidence supports the link between the administration of corticotrophin and fetal body mass, maturity and age of neonates. PMID- 15537245 TI - [Biochemical markers of preterm delivery]. AB - The paper presents the current status of knowledge about pathology of preterm delivery with presentation of biochemical markers diagnostically useful in prediction of this pathology. Infectious cause of preterm delivery and usefulness of infection diagnostic markers in it's prediction are discussed. Diagnostic potential of immunological inflammatory markers such as IL-6, TNF-alfa and fibronectin and oestriol are emphasised in the context of recently published data. The present status of knowledge doesn't allow for pointing of the most sensitive and specific single predictor of preterm delivery. The most promising are multi-marker investigations performed in pregnancy with risk factors. PMID- 15537246 TI - [Immunological aspects of premature termination of pregnancies]. AB - In the presented article we report hypotheses, opinions and confirmed results of investigations concerning immunological mechanisms, that play a role in the physiological pregnancy maintenance, natural reproduction failure and artificial reproductive techniques (ART) as well as abilities to diagnose immunological disorders. We pointed the important role of histocompatibility antigens such are class I and class II major histocompatibility antigens with their soluble forms (HLA-I; HLA-II), DQ-alpha and DQ-beta alleles as well as non classic antigens: HLA-G, HLA-C, HLA-E, in modulation of cellular and humoral response, which represent potential alloimmunological mechanism for pregnancy losses. The accurate report of immunological mechanism for recurrent spontaneous abortions caused by maternal production of phospholipid antibodies in antiphospholipid syndrome (APS) or other antibodies implicated as etiological factors in reproductive autoimmune failure syndrome (RAFS) has been demonstrated. We present the utility of identification of, so-called, reproductive immunophenotype, where an increase of some serum lymphocyte subpopulations anticipating pregnancy loss is frequently observed. In the study we describe recommendations and diagnostic capabilities for detection of immunological disturbances in patients undergoing evaluation for repeated pregnancy loss, which could be helpful in choosing the form of therapy. PMID- 15537247 TI - [Preterm deliveries after in vitro fertilization treatment of infertility]. AB - Wide access to the assisted reproductive technics as a tool in the treatment of infertility arisen many questions regarding the outcome of the pregnancy, its possible complications and the neonatal outcome in women after IVF treatment. We completed the literature search in order to present this subject. It seems that the increased rate of spontaneous abortions and preterm deliveries in this group is caused by such factors as: maternal age, chromosomal, hormonal and uterine abnormalities and immunological factors. The incidence of preterm deliveries in the multiple gestation after IVF group is similar to the one observed in the spontaneous pregnancy group. PMID- 15537248 TI - [Preterm birth in multiple pregnancy]. AB - The incidence of twin and higher-order multiple gestations has increased significantly over the past 15 years primarily because of the availability and increased use of ovulation-inducing drugs and newly developed assisted reproductive technologies. Physicians now must deal with a previously unknown type of multiple gestation, that is, the iatrogenic multiple pregnancy. The most significant and common complication of multiple pregnancy is preterm labor resulting in preterm delivery. Perinatal morbidity and mortality are affected by gestational age and weight at delivery as well as by the number of fetuses. Patient education, risk assessment, serial cervical evaluation by manual or transvaginal ultrasound examinations, specialized antepartum clinics will be use to prevent preterm birth in multiple gestations. PMID- 15537249 TI - [Delivery by natural birth or caesarean section in preterm labour]. AB - The paper describes current opinions concerning the management in the imminent preterm labour and the mode of delivery in different periods of pregnancy before term. PMID- 15537250 TI - [The significance of HLA-G for miscarriages]. AB - In the article, we described major histocompatibility antigens and their distribution in the feto-maternal junction. In particular we have taken into consideration HLA-G, its structure and isoforms. Further on, we cited opinions about their role during reproductive processes, creation of syncytiotrophoblast, remodeling of spiral arteries, growing and maturation of placenta. The lack of correct function of HLA-G can lead to disorders in ovum implantation, hormonal and nutritional function of placenta which can cause reproductive failure, including abortions, intrautrine growth retardation, intrauterine fetal demise, preeclampsia. The genotype HLA-G is potentially related to habitual abortions. PMID- 15537251 TI - [Effect of maternal pharmacological treatment of preterm delivery for neonatal condition]. AB - In article the fetal and neonatal side effects of tocolytic agents were presented. Described the cardiovascular; pulmonary metabolic, neurological and others complications during administration of magnesium sulfate, prostaglandin synthesis inhibitor, calcium channel blockers and oxytocin antagonist. PMID- 15537252 TI - [Congenital cytomegalovirus infection--clinical characteristics including prenatal diagnosis]. AB - Congenital infections are the important cause of morbidity and mortality of newborn infants. Among them, cytomegalovirus (CMV) infection is the frequent congenital injection worldwide with the incidence 0.3-2.2% of live births. The virus can be transmitted to the foetus through primary (in 30 to 40%) or secondary (rare) maternal infection. 90% of infants with congenital infection have no signs or symptoms at birth. The other 10% will be symptomatic and about half of the will have the classic stigmata of CMV inclusion disease. Ten to thirty percent of the severely affected infants will die, more then 90% of survivors will have neurological handicaps, sensorineural hearing loss, chorioretinitis. Ten to 15% of children asymptomatic at birth have a risk of abnormal development and 10% develop hearing loss during the first years of life. Prenatal detection of congenital disease by ultrasound examination is not sufficient, but prenatal diagnosis can be confirmed by amniocentesis (culture of amniotic fluid and detection of virus DNA by quantitative polymerase chain reactions). However, the number of mothers who underwent amniocentesis and the broad case definition used to define symptomatic disease is still not adequate. Because there is currently no treatment for prenatal infection, early diagnosis of congenital infection during neonatal period is important (the possibility of antiviral treatment of children with symptomatic congenital disease and long-term follow-up). The best hope for eliminating congenital CMV infection is to prevent prenatal infection: with no vaccine, good hygiene and handwashing should be promoted. PMID- 15537253 TI - [Fetal and neonatal congenital malformations and premature delivery at a tertiary center]. AB - At the Dept for Diagnosis and Prophylaxis of Congenital Malformations at the Polish Mother's Memorial Hospital in Lodz, in 1994-2001 genetic sonogram + fetal echocardiography have been performed in 1861 women with fetal malformations. In majority there were CNS malformations 25% and congenital heart defects 23%; other malformations consisted of 52%. In 1999-2001 in 407 cases follow-up was available (100%). In 10.8% there was an intrauterine demise, in 40% there were spontaneous deliveries, in 49.2% cesarean sections were performed. In a group of vaginal deliveries (40%), there was 16.2% of premature deliveries < 37 wks and in 23.8% the deliveries were at term (> 37 wks). In a group of CS (49.2%) there were premature babies (< 37 wks) in 21.0% and term neonates (> 37 wks) in 28.2%. There were 37.2% preterm neonates (< 37 wks) with prenatal diagnoses of congenital malformations. In this group there was 64% of neonatal deaths. In group of term neonates with prenatal diagnoses of congenital malformations during the same time there was 38% of deaths. CONCLUSIONS: 1. Fetal/neonatal malformations in current perinatology is a problem of increasing significance for the society. 2. There is a possibility of an early detection of congenital malformations in obstetrical screening and proper final diagnoses at the tertiary center 3.37% of preterm neonates at the tertiary center among population with congenital malformations and 64% of mortality in this group--create and important clue for further investigation of this subject. PMID- 15537254 TI - [Long-term tocolysis--sense and nonsense in current practice]. AB - Preterm delivery is still a major medical and social problem in perinatology. Despite many efforts the appliance of efficient prevention methods is still not possible. The treatment and inhibition of uterine contractions is widely used in clinical practice. The use of tocolytic agents in arresting premature labor is justified by gaining time which is necessary for applying antibiotics and steroids. Short-term tocolysis, ordinarily used between 22 and 34 weeks gestation, prolongs pregnancy and usually delays the delivery by about 48 hours. There are two prevailing options to follow successful inhibition of preterm contractions. The first one recommends intravenous treatment in cases with recurrent premature contractions, the other recommends long-term tocolysis even in cases with no any threatening symptoms. In such cases many obstetricians apply oral beta-Adrenergic agonists, although most of literature data and clinical practice do not confirm good results in preventing premature labor long-term tocolytic therapy does not improve fetal condition and does not prolong the pregnancy. It is also not meaningless that during this therapy maternal adverse effects may occur. Taking the above into consideration long-term tocolysis cannot be recommended as the routine treating procedure. PMID- 15537255 TI - [Nonobstetrical surgical procedures and risk of preterm labour]. AB - Some nonobsterical surgical procedures are described and their influence on pregnant women. The principles of laparoscopy in pregnancy were also shown. PMID- 15537256 TI - [Demographic and socioeconomic risk factors of preterm delivery]. AB - Preterm labour is the main reason of fetal and neonatal deaths. The pathology and etiology of preterm labour are still unknown. The aim of this study was to estimate the demographic and socioeconomic risk factors of premature labour. The study was carried out on 158 pregnant women with preterm labour and rupture of membranes and 120 women with preterm without ruptured membranes. This study determine that age (both young and advanced), low education level, and absent or late prenatal medical care have association with the risk of preterm delivery. PMID- 15537257 TI - [Cervical ultrasound assessment for predicting the risk of premature delivery]. AB - BACKGROUND: Cervix is very important structure that separates the fetus from external environment, it can be visualized very precisely by transvaginal ultrasonography. Cervical length in the pregnancy is one of the predictors of preterm delivery and diagnosis of cervical incompetence. MATERIALS & METHODS: We have investigated 119 pregnant women with diagnosis of preterm labor(n-62), with preterm labor complicated by diabetes mellitus(41) and hypertension (16). Transvaginal sonography was used to measure cervical length between 23 and 29 weeks gestation. Ultrasound examination was performed using 2000 with 5.0 MHz transvaginal transducer. The following measurements were obtained: endocervical canal length between the internal and external os, funneling of the cervix, funnel width, funnel length. RESULTS: The mean endocervical length was 16.3 +/- 0.5 mm in pregnancies delivered before term (n = 19) and 30.1 +/- 0.6 mm in pregnancies delivered at term (n = 100). The mean endocervical length measured in pregnant women without pregnancy complications other than preterm labor was 15.2 +/- 0.6 mm, in women with diagnosis of preterm labour complicated by diabetes mellitus was 16.7 mm + 1.0 and complicated by hypertension 15.9 mm + 0.8. No significant relationships were shown between length of the cervix in pregnancy complicated by diabetes mellitus, hypertension and uncomplicated pregnancy. The mean endocervical length in diabetic pregnancy was 34.7 +/- 4.7 mm; pregnancy with hypertension 32.9 +/- 5.8 mm and uncomplicated pregnancy 35.6 +/- 3.8 mm. CONCLUSION: Cervical length measured by ultrasonography between 23 and 29 week of pregnancy is very important predictor of preterm delivery. We found significant relationships between cervical length in pregnancies delivered before and at term. No significant relationships were shown between the length of the cervix and investigated pregnancy complications. PMID- 15537258 TI - [Sex during pregnancy and risk of premature labour]. AB - Sexual life in pregnancy may be connected with preterm delivery. This fact, which is observed in clinical practice, is not confirmed by, references. There are no modern, well conducted trials about the risk of premature labour as a result of sexual activity. The concentration of fetal fibronectin is false, when the examination is performed before 24 hours from the last sexual intercourse. PMID- 15537259 TI - [Sexuality in pregnant women]. AB - The aim of the study was to evaluate the effects of pregnancy on women's sexuality. The studies were conducted using questionnaires to interview 120 women from 35 weeks' of gestation or just after the delivery. There were no medical contraindications for sexual intercourse in pregnancy in this group. More than half of respondents reported a decrease in libido during gestation. A reduction in coital frequency, oral intercourse, gratification of coitus, caress of breasts or genitals and erotic dreams occurred during pregnancy. Lateral position was mainly used during coitus in pregnancy. Most of women didn't discuss the problems of sexual activity in pregnancy with their obstetricians. PMID- 15537260 TI - [Effectiveness of cervical cerclage and pessary in the treatment of isthmo cervical insufficiency]. AB - The aim of this study was comparison of gestation, labour, puerperium and condition of newborns in women with cervical incompetence treated with cerclage or pessary. The interval from treatment to delivery was longer in women with pessary. In women who had cervical cerclage tocolysis was applied more frequently. Among patients with pessary kolpitis was found more often. Gestational age at delivery, the way of a labour, condition of newborns and complications during puerperium didn't differ significantly between both groups. PMID- 15537261 TI - [Results of amnioinfusion in pregnancies with oligohydramnios and non-ruptured fetal membranes]. AB - Authors analysed results of amnioinfusion in the group of 65 pregnant women with oligohydramnios and non-ruptured fetal membranes. Amnioinfusions were the most efficient in the pregnancies with oligohydramnios and asymmetric hypotrophy, idiopathic oligohydramnios or oligohydramnios with pathological FCG. The group with oligohydramnios and fetal malformations showed the worst results of the treatment. The findings confirm mainly the diagnostic character of the method before the end of 25 week of pregnancy because during period the oligohydramnios usually occurs together with lethal fetus malformations. The amnioinfusion in the cases of fetal hypotrophy and pregnancy-induced hypertension seems to be the prophylactic procedure. PMID- 15537262 TI - [Analysis of preterm labours at the Obstetrics and Gynecology Department of the Specialist Hospital in Jaslo, during the period of 1998-2001]. AB - There were 128 preterm labours analyzed, all between 22nd and 37th week of gestation. This equals 2.71% of all deliveries which occurred taken place in the Obstetrics and Gynecology Department of the Specialist Hospital in Jaslo, during period of 1998-2001. In the group discussed, there were 16 twin--labour and one triplet--labours, hence the number oj infants was amounted to 147. The fetuses and infants with birth weight below 2500 g were divided according to weight range each 500 g. Next, the following aspects were analyzed: rate of premature births, the way of pregnancy completion, indications to cesarean section, condition of the newborn within 1 minute of life according to Apgar's score, birth weight and perinatal mortality. The decreasing number of babies with extremely low birth weight during analysed period was stated. There is also a low average percentage of premature deliveries (2.69%). In addition, there is a growth of sections rate and significant decrease in infants death. All this is concerned with increasing effectiveness of perinatal care. PMID- 15537263 TI - [Relationship between cervical infections and premature rupture of membranes]. AB - Preterm delivery is still one of the most important problems of the modern obstetrics. In our country frequency of preterm delivery is 7.2-8.4% of all deliveries. Although preterm birth is the leading cause of perinatal morbidity and mortality, the diagnosis and appropriate treatment of preterm labor remains an unresolved clinical problem. A growing body of evidence supports the view that preterm labor is a syndrome with multiple causes and the urogenital infections play a very important role in the pathogenesis of preterm birth. The study analyses relationship between results of microbiological examination of cervical smear and preterm rupture of membranes. PMID- 15537264 TI - [Association between genital mycoplasmas and risk of preterm delivery]. AB - The main aim of this study was to determine the prevalence of maternal colonisation by genital mycoplasmas during pregnancy and to assess its association with preterm delivery. The cervical swabs of 179 pregnant women between 8 and 16 week of gestation were cultured for Mycoplasma hominis and Ureaplasma urealyticum. Clinical details and perinatal outcomes were documented. Of a total of 179 women enrolled, 68 (38.0%) had positive cervical culture for genital mycoplasmas; 30 (16.7%) for M. hominis and 38 (21.3%) for U. urealyticum. Preterm delivery occurred in 21 (11.7%) women. Genital mycoplasmas were found more frequently in group with preterm delivery as compare to women who delivered at term (p = 0.05) and its presence at lower genital tract at early pregnancy was a risk factor of preterm delivery. Early pregnancy screening for genital mycoplasmas and following treatment may reduce the rate of preterm deliveries. PMID- 15537265 TI - [Bigeminal pregnancy complicated with twin-to-twin transfusion]. AB - Twin-to-twin transfusion syndrome is one of the complications during bigeminal pregnancy usually seen between 17 and 31 weeks of gestation. The most typical symptoms of twin-to-twin transfusion are ascites, hydropericardium and generalized hydrops of donors fetus. Characteristic is also difference of body weight over 15% and hemoglobin level more than 5 g/dl between twins. The mortality rate of donors is very high approaching 50%. In this case report we found some features of twin-to-twin transfusion: time of delivery--33 weeks of gestation, mode of delivery--cesarean section, generalized hydrops of the donor. The clinical status of donors was critical--Apgar score 1-1-1. The hemoglobin level was extremely low--3.2 g/dl. The rescue treatment was not successful and donors twin died. Clinical problems of second twin was also common: polycythaemia, hypoglycaemia and hyperbilirubinaemia. PMID- 15537266 TI - [Twin birth weight discordance and risk of preterm birth]. AB - Our purpose was to determine whether birth weight discordance is a risk factor for preterm birth of twins. Maternally linked 1970-2000 Kutno Hospital birth certificates were used to analyze gestations resulting in live twins. The degree of discordance correlated strongly with risk for live preterm birth but only for discordances > 20%. Twin birth weight discordance has demonstrated to be a risk factor for preterm birth. The effect was found particularly with discordance > 30%. PMID- 15537267 TI - [Conjoined twins (Monstra duplicia)]. AB - The Authors present a delivery of conjoined twins--thoracopagus, of an 18 years old primigravida. A pregnant woman was admitted to delivery room with full cervical dilation. She underwent ultrasonography examination which showed bigeminal pregnancy--two fetus in breech presentation with the suspicion of conjoined twins. Ultrasonography measurements of fetus corresponded to gestational age of 23 weeks. The heart rate of the first fetus was about 40 per minute, the heart rate of the second fetus was undetectable. She underwent delivery through manual extraction of the fetuses. Both fetuses were female, without any signs of life. The result of the autopsy was: Hemithoracogastropagus holoacardius immaturus. PMID- 15537268 TI - [Cervical incompetence in twin pregnancy--case report]. AB - The paper presents a case of a 29-years old primipara with the cervical incompetence diagnosed in 22nd week of twin gestation. Cervical cerclage was performed with a cervical dilatation about 7 cm. The pregnancy was prolonged for 84 days. Caesarean section was performed in 34th week of gestation. The mother with healthy newborns was discharged from the Department on 5th day after delivery. PMID- 15537269 TI - [Fetal arrhythmia--own experience]. AB - Experience in diagnosis and treatment of fetal arrhythmia of Prenatal Cardiology Clinic in the Children's Memorial Health Institute (until 2001), and then the Perinatology and Perinatal Cardiology Clinic of the 2nd Department of Obstetrics and Gynecology in the Medical University of Warsaw are discussed. The author described physiology and pathophysiology of the fetal conduction system, diagnostic and therapeutic methods of fetal arrhythmia. We retrospectively analyzed 130 fetuses with arrhythmia. Median time of diagnosis was 30 weeks, median maternal age 28 years. Extrasystole was the most common type of fetal arrhythmia (83 fetuses). 19 of them had to be closely followed up, because there were structural anomalies in 6 (7.2%) of them, and progression of arrhythmia (bradycardia, supraventricular tachycardia or atrial flutter) in 17. Supraventricular tachycardia was diagnosed in 17 fetuses (12--SVT, 5--atrial flutter AF), and all of them were treated in utero. Transplacental therapy with digoxin, flecainide, sotalol and amiodarone was used, and transumbilical emergency treatment with adenosine was introduced in 2 fetuses with severe hydrops. Complete heart block (CHB) was in 12 fetuses, in 8 with severe heart defects, in 4 was caused by anti-Ro antibodies. We concluded that extrasystole, although in majority were benign condition, in 15% fetuses needed to be followed up, due to heart defect or more serious arrhythmia. Type of rhythm disturbances can change during pregnancy. SVT and AF could be successfully treated transplacentally. All cases with CHB and heart defect died in utero or shortly after birth. Follow up of fetuses with CHB without heart defect seemed to be much better. PMID- 15537270 TI - [Intraamniotic surfactant supply as RDS prevention]. AB - The importance of steroids given antenatally to the mothers in prevention of the respiratory distress syndrome (RDS) is unquestionable. Also intra tracheal surfactant application in newborn is proven method of prevention and treatment of RDS. However both options have some limitations and new methods useful in prevention of RDS are still needed. The aim of the study was to evaluate the efficiency and safety of the procedure of the intraamniotic surfactant supply as RDS prevention. Natural surfactant (Alveofact - Boehringer Ingelheim) has been given to 15 women at 24-32 weeks of pregnancy, two hours before expected childbirth (mainly cesarean section). To 8 of these women corticosteroids have been earlier administered. Just before surfactant injection amniotic liquid samples were taken to confirm lung immaturity and the patients were administered Aminophilline intravenously to provoke fetal breathing movements. Surfactant has been administered through the needle under direct ultrasound guidance into the amniotic cavity as close as possible to the fetal mouth and nose. Patients with the evidence of chorionamnionitis and fetal malformations were excluded from the study. No complications were observed during and after the procedure. None of the newborns had symptoms of severe asphyxia, the birth weight comprised between 670 1650 g (mean 1207). There was radiological evidence of RDS in two newborns and in 7 mechanical ventilation was needed. Of 15 newborns 13 survived (86.6%). There was no need for postpartum surfactant therapy in any case. Some authors expressed their view that this method is promising and further studies are desirable. In our study surfactant has been injected into the amniotic cavity shortly before childbirth, and Aminophilline has been administered intravenously in order to provoke fetal breathing movements before surfactant injection. The safety of the procedure was confirmed and the results of intraamniotic surfactant supply seems to be favourable to newborns. Small number of cases does not allow to draw any far-reaching conclusions. Still our preliminary results are encouraging and the study should be continued. PMID- 15537271 TI - [Effect of bethametasone on blood cell count and C-reactive protein in patients with threatened preterm delivery]. AB - The objective of the study was to determine changeableness of blood cells count, hemoglobin (HGB) and hematocrit (HCT) within the peripheral blood of patients who received betamethasone during treatment of threatened preterm delivery. 65 patients at increased risk of preterm delivery received intramuscular injections of betamethasone 3 x 4 mg/day for fetal lung maturity and to decrease the risk of CNS trauma in newborns. RBC, HGB, HCT, WBC, neutrophils count and CRP level were measured before treatment and on the 1th, 2nd and 7th days after first dose of betamethasone. The steroid administration caused a significant decrease in RBC, HGB and HCT: The lowest levels were noticed on 2nd day. Statistically significant changes were observed between before and 1st day (p < 0.0001), before and 2nd day (p < 0.0001), before and Ist week (p < 0.05), 1st day and 1st week (p < 0.006), 2nd day and Ist week (p < 0.005). The level of WBC and neutrophils were increased. Their peak levels were noticed on 1st day. Statistically significant changes were observed between before and 1st day (p < 0.0001), before and 1st week (p < 0.0003), 1st day and 1st week (p < 0.0001), 2nd day and 1st week (p< 0.001). After 3 days there was a significant decrease of WBC and neutrophils count. CONCLUSIONS: Intramuscular administration of 12 mg betamethasone per day caused decrease of the RBC, HGB and HCT account and elevation of the WBC and neutrophil count. All these parameters normalized after about 7 days. PMID- 15537272 TI - [Pregnancy and travel]. AB - The special problems of travel during pregnancy have become clinically important as more women are traveling to remote places for business or recreation. Many issues need to be considered in providing health advice to the pregnant traveler. Health practitioners should be able to assist in the careful medical preparations that will ensure the safety of the pregnant woman and fetus during travel. Journey may increase the risk of pregnancy complications because of inherent physiological changes or exposure to environmental hazards and infectious disease. Usually standard interventions are not recommended during pregnancy. In pregnancy, the maternal and fetal benefits of protection from disease and its complications must be carefully balanced against the small, often theoretical risk, of adverse effects following immunization. In some circumstances the best advice may be to postpone travel if effective preventive measures against disease cannot be undertaken and access to medical care is uncertain. PMID- 15537273 TI - [Successful pregnancy in a patient with chronic renal insufficiency treated with repeated hemodialysis]. AB - A case of successful pregnancy in a patient with chronic renal insufficiency is presented. Appropriately conducted conservative treatment with repeated haemodialysis were decisive for successful outcome of pregnancy. PMID- 15537274 TI - [Caesarean section in preterm labour]. AB - The aim of this study was to estimate complications of pregnancy, indications to caesarean section, complications after caesarean operation and perinatal mortality of newborns who were delivered before the 37th week of gestation by caesarean section. The retrospective analysis of the total of 2693 deliveries (years 1991-2000) by caesarean section in the Department of Pregnancy and Labour Pathology in Pomeranian Academy of Medicine has been carried out. 590 women whose delivered before the 37th week of pregnancy have been chosen for subsequent examination. The patients have been divided into three groups: group I--patients with pregnancy completed before the 28th week, group II--patients with pregnancy completed between 29th and 32nd week, and group III--patients with pregnancy completed between the 33rd and 36th week. Obstetrical anamnesis, complications of pregnancy, indications to caesarean section and postoperative complications as well as newborns perinatal mortality have been analysed. Among women who experienced abortions, premature deliveries caesarean section a higher rate of caesarean section during subsequent premature delivery was stated. The increase of caesarean sections rate in premature deliveries after the 29th week of gestation is associated with a considerable decrease in newborns, mortality. PMID- 15537275 TI - [Prostaglandin E2 gel for labor induction in preterm pregnancy]. AB - OBJECTIVES: Endocervical prostaglandin E2 gel administration is widely accepted mean of induction of labor in pregnancy at term. Further investigation is needed to assess its usage in pregnancy below 37 weeks' gestation. The aim of this study is to analyse the course of labor, its complications and neonatal outcome in preterm deliveries induced with endocervical dinoprostone. RESULTS: We conducted a retrospective study of 22 preterm deliveries induced with dinoprostone. A control group consisted of 26 pregnancies at term. The incidence of cesarean sections was comparable in both preterm and term deliveries (45 vs. 41%, p = 0.38). We found no differences in characteristics of indications of operative delivery, duration of particular stages of labor; time from dinoprostone administration to the beginning of 1st stage of labor, estimated blood loss. 1 and 5-minute Apgar scores were similar in both groups. CONCLUSIONS: Endocervical prostaglandin E2 gel administration seems to be a safe method of induction of labor in preterm pregnancy. PMID- 15537276 TI - [Use of misoprostol in the termination of missed abortion]. AB - The efficiency of misoprostol in the termination of missed abortion was estimated. The group of 66 patients aged 19 - 37, who received 400 microg of vaginal misoprostol for termination of missed abortion was subjected. The overall success rate for a complete abortion was 30.3%, for incomplete abortion 25.8% and in a group of 27.2% women there was no need for cervix canal dilatating during the excochleation of uterus cavity. 16.7% cases failed. The results obtained in this study led us to the conclusion that 400 microg of vaginal misoprostol can induce the termination of missed abortion or dilatation of the uterus' canal. PMID- 15537277 TI - [Curettage of uterine cavity in late puerperal bleeding]. AB - Sixty eight (68) female patients were subjected to curettage caused by late puerperal bleeding. The above group is considered as 0.8% of all the female patients who delivered children during the period of research. The most frequent causes of bleeding were: abnormal desquamation of decidua (48.5%), residua of secundines (32.4%), and endometritis (14.7%). Prior to curettage, every patient was subjected to ultrasonographic examination. The suspicion of residual secundines, found in ultrasonographic examination was later confirmed in 36.8% of patients. No significant interdependence between the results of ultrasonographic and histopathological examinations, were noted. The disturbances in natural process of decidua excretion were rarely diagnosed in ultrasonographic pictures (3%) when puerperal bleeding required curettage of uterine cavity. PMID- 15537278 TI - [Genetic amniocentesis--characteristic of patients, indications, outcomes, complications]. AB - The indications, outcomes and complications of genetic amniocentesis in a group of 358 women were undeped. The most frequent indication for amniocentesis was maternal age over 35 years. Chromosomal abnormalities were identified in 21 cases (5.8%)--most of them were chromosomal inversions. There were no neural tube defects. In 6 (1.6%) cases the cell culture of amniotic fluid was unsuccessful. The level of alpha-fetoprotein was lower in fetuses with chromosomal abnormalities. 2 (0.5%) patients lost pregnancy after procedure, 4 (1.1%) had amniotic fluid leakage and 1 (0.2%) had pain in hypogastrium after amniocentesis. PMID- 15537279 TI - [Evaluation of risk factors for neonatal infection in preterm infants--own experience]. AB - Preterm birth is one of the major problems in perinatology today. It is associated with increased risk of neonatal infection and related neonatal mortality. In this study we evaluated some perinatal risk factors for neonatal injection. We found an association of neonatal infection with low gestational age, low birth weight, low Apgar score, intraamniotic injection, maternal leukocytosis, prolonged ruptured membranes and antenatal use of steroids and tocolysis. We conclude, that appropriate diagnosis, treatment and prophylaxis of intrauterine infections may reduce the incidence of infections among preterm babies. PMID- 15537280 TI - [Maternal serum procalcitonin concentrations in preterm labour complicated with premature rupture of membranes]. AB - The aim of this study was the evaluation of serum procalcitonin (PCT) concentrations in pregnancy complicated by premature rupture of membranes (PROM) before 36 weeks of gestation. The study group consisted of 42 women with PROM between 25. and 35. week of pregnancy. The control group comprised 38 patients between 24.-35. weeks of pregnancy in acute phase of imminent preterm labor with intact membranes. All of them, after successful treatment in the hospital, delivered near term. In both groups serum PCT concentrations were higher than in healthy patients without pregnancy as compared to literature data. It was also stated that in women with PROM before 36. week of gestation serum PCT concentrations are significantly higher than in pregnant women in acute phase of imminent preterm labor with intact membranes who after successful treatment deliver near term. PMID- 15537281 TI - [Longitudinal study of children with bronchopulmonary dysplasia treated with disodium cromoglycate]. AB - In order to improve the quality of life of children born prematurely, who developed chronic lung disease, clinical trials of drugs of different origin are undertaken. The aim of the work was the evaluation of the efficacy of disodium cromoglycate in the treatment of bronchopulmonary dysplasia in children. We retrospectively studied 15 infants with bronchopulmonary dysplasia (BPD) hospitalised in the Infant Care Department of Children's Health Memorial Institute from 01.01.1997 to 01.02.2000. All babies were premature (25-30 weeks of gestation) with LBW or VLBW A control group of 11 babies with BPD, matched for birth weight and gestational age, who did not have disodium cromoglycate therapy were also studied. Recurrent obturative bronchitis and bronchial hyperresponsiveness were stated in all cases in both groups. Disodium cromoglycate was administered in all babies in the study group. Inhaled corticosteroid (Budesonide mite) was given in 10 cases, for a short period of time, due to severe obturative bronchitis. Babies in the control group were treated with systemic and inhaled corticosteroids. Results of our trial compared with the log-rank and chi2 test show statistically, significant differences in the regression of obturative bronchitis (log-rank = 4.35, p < 0.0001) and normalization of capillary blood-gas examination (log-rank = 3.777, p < 0.0002) in favour of the studied group, treated with disodium cromoglycate. PMID- 15537282 TI - [Evaluation of CD3+/TCR-alpha-beta and CD3+/TCR-gamma-delta lymphocytes in umbilical cord blood]. AB - The neonate's immune system is immunocompetent, but because of the "virgin" status, the immune system do react differently to antigens and is less efficient. The aim of the study was to determine the relative and absolute numbers of CD3+/TCR alpha beta and CD3+/TCR gamma delta lymphocytes in the umbilical blood. The examinations comprised 26 full term neonates and 41 preterm ones. The immunophenotyping analysis was performed using an FACScan flow cytometer and anti TCR alpha beta, anti- TCR gamma delta monoclonal antibodies (Becton Dickinson). The mode of delivery did not influence significantly the value of CD3+/TCR alpha beta lymphocytes. The elective cesarean section influenced significantly on the decrease of relative and absolute number of CD3+/TCR gamma delta lymphocytes in neonates. Low absolute number of CD3+/TCR gamma delta lymphocytes was observed in the preterm infants born after preterm placenta separation and preterm rupture of membranes. The immune system of the newborn infant is immature and hyporesponsive when compared with adults. Neonates are more susceptible to infection than adults, and exhibit more severe or prolonged symptoms when infected. PMID- 15537283 TI - [Sexually transmitted infections in sexually abused children]. AB - The sexually transmitted infection (STI) in an infant can be an early consequence of child sexual abuse (CSA). The risk of acquisition of a STI depends, among others, on the prevalence of these infections in the abusing population and the type of sexual assault. As the detection of sexually transmissible organism can be an unique evidence of CSA, only the best standardized methods should be used for the diagnosis of STIs in children. STIs are seen in less than 10% of abused child and in a half of them are asymptomatic. The relevance of STIs for the diagnosis of CSA should be assessed taking into consideration the age of a child, a type of sexually transmissible organism and its localization. Generally, postnatally acquired gonorrhoea, syphilis and genital Chlamydia trachomatis infection in a child more than 2-year old are most clinically evident for CSA. Infections with HPV, HIV and HSV can also be important. PMID- 15537284 TI - Speciation of aluminum in drink samples by 8-hydroxyquinoline loaded silylanization silica gel microcolumn separation with off-line ICP-MS detection. AB - A technique using a flow injection microcolumn separation coupled with ICP-MS detection has been developed for the speciation of Al in drink samples. The retention behaviors of different Al species were studied with 8-hydroxyquinoline (8-HQ) loaded silylanization silica gel as the packing material and inorganic acid (HNO3) as the elution. The results indicated that in a pH range of 5.0 to 8.0, all labile monomeric Al species were retained on the microcolumn while nonlabile monomeric Al species were directly passed through the column. Various Al species after separation were detected by ICP-MS. The detection limit of 0.2 ng mL(-1) and a relative standard deviation (RSD) of 4.2% at 10 ng mL(-1) (n = 11) were achieved, and the recoveries for the spiked samples were 95-108%. The proposed method has been applied to the analysis of Al species in tea infusions, coffee, and tap waters with satisfactory results. The results obtained by this method were compared with that obtained by the cation exchange microcolumn separation and ICP-MS detection system, and some valuable conclusions were drawn. PMID- 15537285 TI - Development of multiclass methods for drug residues in eggs: silica SPE cleanup and LC-MS/MS analysis of ionophore and macrolide residues. AB - A method was developed that is suitable for screening eggs for a variety of nonpolar residues in a single procedure. Residues are extracted by silica solid phase extraction (SPE). Analysis is conducted via reverse-phase gradient liquid chromatography, electrospray ionization, and tandem ion trap mass spectrometry. For screening purposes (based on a single precursor-product ion transition) the method can detect ionophore (lasalocid, monensin, salinomycin, narasin) and macrolide (erythromycin, tylosin) residues in egg at approximately 1 ng/mL (ppb) and above and novobiocin residues at approximately 3 ppb and above. Conditions are described for confirmatory analysis based on multiple ions in the product ion spectrum. The extraction efficiency for ionophores was estimated at 60-85%, depending on drug. Recovery of macrolides and novobiocin was not as good (estimated at 40-55% after a hexane wash of the final extract was included), but the method consistently screened and confirmed these residues at concentrations below the target of 10 ppb. The method was applied to eggs from hens dosed with each drug individually. Lasalocid was found to have the highest probability of detection in eggs based on its high ionization efficiency and higher rate of deposition relative to the other drugs. The method is part of a larger scheme to provide surveillance methods for a wide variety of drug residues in eggs. PMID- 15537286 TI - Fast screening method for volatile compounds of oak wood used for aging wines by headspace SPME-GC-MS (SIM). AB - A headspace solid-phase microextraction (HS-SPME) method is proposed for analyzing the main volatile components from a sensory standpoint (furfural, oak lactones, eugenol, vanillin, and syringaldehyde) present in nontoasted and toasted oak wood of different origins. To maximize the yield of compounds extracted from wood chips and to obtain a good precision of the method, the most important variables affecting HS-SPME have been studied. The best results were obtained when the sample was heated at 70 degrees C and the headspace extracted for 40 min with a DVB/CAR/PDMS fiber, which gave the overall best recovery. The values for the repeatability ranged from 6.4 to 7.8%, and those for the reproducibility from 5.4 to 8.7%. The precision of the results obtained makes the proposed technique appropriate for its use in characterizing oak wood samples of different origins and in the selection of the most suitable oak wood to age wines and spirits, on the basis of the chemical composition of the wood samples. PMID- 15537287 TI - Fluorometric sucrose evaluation for sugar beet. AB - Sucrose is the economic product from sugar beet. Disease resistance is often available in low-sucrose genotypes and, prior to the deployment of such novel genes as available into the cultivated spectrum, selection for increased sucrose content is required during introgression. The objective of this work was to evaluate a relatively rapid and inexpensive enzymatic-fluorometric microtiter plate assay for sucrose quantification in sugar beet root dry matter, both for progeny testing in the greenhouse and for evaluation of field-grown mother roots. As determined using HPLC, sucrose content in diverse populations of sugar and table beet assayed over various developmental stages ranged from 0.213 to 2.416 mmol g(-1) of dry matter, and these values were used as references for both refractometry and enzymatic-fluorometric assay. As expected, refractometric analysis generally overestimated sucrose content. Enzymatic-fluorometric analyses were reasonably well correlated with HPLC results for young greenhouse-grown root tissues (R2 = 0.976), and less so with older field-grown roots (R2 = 0.605), for unknown reasons. Enzymatic-fluorometric assays may be best deployed for progeny testing of young seedlings. PMID- 15537288 TI - Liquid chromatography-electrospray tandem mass spectrometry of cis-resveratrol and trans-resveratrol: development, validation, and application of the method to red wine, grape, and winemaking byproducts. AB - The application of liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) was investigated for the analysis of trans-resveratrol in red wine, grape skin, grape pomace, and winemaking byproducts. Chromatographic elution performed under isocratic reversed-phase conditions using a C18 narrow bore LC column allowed retention times lower than 12 min to be obtained. Qualitative analysis was performed in negative-ion (NI) full-scan and product-ion scan acquisition modes, whereas method validation in terms of linearity, detection limits, accuracy, and robustness was carried out under NI selected reaction monitoring conditions. The matrix-matched detection limit was calculated in the low parts per billion range (10 microg/L). Results of the application of the method to red wine, grape, and winemaking byproduct samples were compared with those obtained using an LC-UV/DAD technique. Determination of trans resveratrol in the samples investigated showed that the highest concentration was found in red wine, whereas wine made from grape pomace contained the lowest content. PMID- 15537289 TI - Determination of methylglyoxal in ruminal fluid by high-performance liquid chromatography using fluorometric detection. AB - There is no reported method for the quantification of methylglyoxal in ruminal fluid. The method reported here is based on the conversion of methylglyoxal to 6 methylpterin, followed by quantification of the resulting pteridinic compound by fluormetric detection using liquid chromatography. Ruminal fluid was collected and preserved with 1 M HCl at -20 degrees C. Cation exchange prior to derivatization was used to eliminate possible interfering peaks. The detection limit of 0.125 microg/mL was calculated. The recoveries were >80%, and the coefficients of variation were <15%. This method has proven to be rugged and accurate for the detection of methylglyoxal concentration in ruminal fluid collected from cows fed diets deficient in degradable intake protein as a marker. Methylglyoxal is produced by ruminal bacteria in response to low nitrogen levels in the rumen. The ruminal methylglyoxal concentration has the potential to be a useful marker to assess ruminal nitrogen status to aid in more accurate diet formulation. PMID- 15537290 TI - Quantification of total furocoumarins in citrus oils by HPLC coupled with UV, fluorescence, and mass detection. AB - Furocoumarins or psoralens represent a class of photosensitizers whose use level is likely to be restricted to 1 ppm in cosmetic products by the EU. A reversed phase HPLC method was developed to separate the 15 main furocoumarins present in citrus oils. Quantification by UV, fluorescence, or mass detectors was compared in terms of linearity and limit of detection. Cold-pressed oils of different citrus species were analyzed using this method. This method could be implemented in quality control laboratories equipped with an HPLC system and a UV diode array detector. Because of possible coelutions, the UV-spectral data should be carefully examined to avoid misleading interpretations of peaks. PMID- 15537291 TI - Enzymatic determination of urea in milk by sequential injection with spectrophotometric and conductometric detection. AB - In this work, an analytical system based on the coupling of gas diffusion separation and sequential injection analysis for urea determination in milk is presented. A versatile manifold that could simultaneously be used for either spectrophotometric or conductometric detection was constructed. The sample and urease solution are sequentially aspirated into the holding coil and sent to a thermoreactor, where urea is enzymatically hydrolyzed by urease and converted into ammonium. This stream merges an alkaline solution at a confluence point where ammonia is formed. Ammonia diffuses through a hydrophobic membrane and modifies the bromothymol blue indicator color, when spectrophotometric detection is used, or changes the conductance of a boric acid solution acceptor stream, when conductometric detection is used. This methodology was applied to the determination of urea in 18 milk samples and the results were statistically comparable with those furnished by the enzymatic recommended procedure. The detection limits were 2.6 x 10(-4) and 2.8 x 10(-5) mol L(-1) for conductometric and spectrophotometric detection, respectively. Repeatability (relative standard deviation, RSD) was better than 3.7% and 2.6% for conductometric and spectrophotometric detection, respectively. PMID- 15537292 TI - Preliminary studies for the differentiation of apple juice samples by chemometric analysis of solid-phase microextraction-gas chromatographic data. AB - A combination of gas chromatography (GC) and chemometrics was evaluated for its ability to differentiate between apple juice samples on the basis of apple variety and applied heat treatment. The heat treatment involved exposure of 15 mL juice samples for 30 s in a 900 W domestic microwave oven. The chromatographic results were subjected to two chemometric procedures: (1) partial least squares (PLS) regression and (2) linear discriminant analysis (LDA) applied to principal component (PC) scores. The percent correct classification of samples were obtained from PLS and LDA in terms of separation on the basis of apple variety and applied heat treatment. PLS gave the highest level of correct classification of the apple juice samples according to both variety and heat treatment, 92.5% correct classification in each case. When LDA was performed on the PC scores obtained from GC analysis, 87.5% and 80% of samples were correctly classified according to apple variety used and applied heat treatment, respectively. PMID- 15537293 TI - Comparison of strategies for extraction of high molecular weight polycyclic aromatic hydrocarbons from drinking waters. AB - Simple, rapid, and inexpensive methods have been developed for the determination of polycyclic aromatic hydrocarbons (PAHs) in drinking waters without interferences from other chemical contaminants by use of two different extraction techniques and analysis by an optimized reverse-phase (RP) high-performance liquid chromatography followed by fluorescence detection (HPLC-FLD) method. The feasibility of SPE (solid-phase extraction) and SPME (solid-phase microextraction) for the determination of PAH in drinking water samples has been evaluated. Several parameters have been studied and optimized for both extraction procedures. The relationship between the nature of the fibers and the quantity of extracted compounds and the effects of organic solvent, salt addition, sampling temperature, and sampling time was studied for SPME. Acetonitrile percentage added to the sample, sample storage conditions (temperature and time), and type of organic elution solvent and elution volume were evaluated for SPE. The results show that both extraction procedures can be used to determine PAHs in drinking waters, but SPE gives better performance (recovery, precision, and quantification limits) for the determination of PAHs in drinking water at the levels established by the legislation. PMID- 15537294 TI - Processing of chromatographic data for chemometric analysis of peptide profiles from cheese extracts: a novel approach. AB - Chemometric analysis of chromatograms plays a fundamental role in characterization of foods or in detection of adulteration. Data for multivariate analysis of chromatographic profiles are usually obtained by visual matching (VM) of peaks, the identities of which, as for peptide profiles from cheese extracts, are often unknown. To avoid the main disadvantages of VM, which is subjective and time-consuming, a novel approach was developed. Fuzzy logic was employed to handle in a systematic way uncertainty in the position of peptide peaks, and chromatograms were processed by a rule-based membership function. Processed data consisted of classes of retention time wherein peak heights were accumulated by using the distance from the center of the class as a weight. The novel approach (fuzzy approach, FA) was compared with VM by using a real data set and by performing multivariate descriptive statistical techniques (principal component analysis, multidimensional scaling, and nonhierarchical cluster analysis). FA provided a fast, reliable, and objective alternative to VM and could be successfully applied for chemometric analysis of chromatographic profiles whenever knowledge of the identity of peaks is lacking or unnecessary. PMID- 15537295 TI - Simultaneous residue measurement of pendimethalin, isopropalin, and butralin in tobacco using high-performance liquid chromatography with ultraviolet detection and electrospray ionization/mass spectrometric identification. AB - A simultaneous residue analysis of pendimethalin, isopropalin, and butralin in tobacco was developed with high-performance liquid chromatography with ultraviolet monitoring and electrospray ionization mass spectrometry (HPLC-UV ESI/MS). The herbicide residues of pendimethalin, isopropalin, and butralin in tobaccos were extracted by ultrasonication with ethyl acetate, followed by a cleanup procedure with gel permeation chromatography. The three herbicides were separated within 15 min using a LiChrosorb 18 column with methanol-10 mmol/L ammonium acetate (85:15, v/v) as the eluent. The limits of quantitation, using HPLC-UV, were ca. 0.05, 0.08, and 0.06 mg/kg for pendimethalin, isopropalin, and butralin, respectively, whereas the overall recoveries ranged from 77.5 to 91.8%. The proposed method has been successfully applied to measure 300 real samples, and the residue profiles of three herbicides in tobacco samples were obtained and evaluated. PMID- 15537296 TI - Bioguided fractionation and isolation of free radical scavenging components from in vitro propagated chinese medicinal plants Dendrobium tosaense Makino and Dendrobium moniliforme SW. AB - This study was performed to investigate the free radical scavenging active components from in vitro propagated medicinal herbs of the genus Dendrobium, namely, Dendrobium tosaense Makino and Dendrobium moniliforme SW, using a 1,1 diphenyl-2-picrylhydrazyl (DPPH) radical antioxidative assay. Seeds of the capsules derived after 12 weeks of hand-pollination germinated asymbiotically (50 74%) on half-strength Murashige and Skoog's (MS) basal medium with 3% sucrose and solidified with 0.9% Difco agar. Active growth in the germinated seedlings was achieved by reculturing on full-strength MS basal medium supplemented with 8% banana homogenate, 8% potato homogenate, 8% coconut water, 1.5% sucrose, and 0.9% Difco agar. Healthy plantlets transferred to plastic trays containing moss or moss and tree fern successfully acclimatized (84-100%) in the greenhouse. Extracts were prepared from plants grown in the greenhouse for a period of 6 months. Methanolic extracts of D. tosaense and D. moniliforme scavenged DPPH at 95.9 and 83.4%, respectively, at a concentration of 0.4 mg/mL. Therefore, methanolic solubles of D. tosaense and D. moniliforme were subjected to bioguided fractionation and separation by column chromatographic methods individually. After chromatographic separation of these crude extracts, the obtained fractions (Dm 1, Dm 2, Dm 3, Dt 1, Dt 2, and Dt 3) were tested for their activity. Among them, fractions Dm 2 and Dt 1 showed significant antioxidant activity by DPPH radical antioxidative assay. Active fractions were purified further by column chromatography and resulted in identification of the antioxidant components alkyl ferulates from D. moniliforme and quercetin from D. tosaense. PMID- 15537297 TI - Improved method to obtain pure alpha-galactosides from lupin seeds. AB - Improvement of a previously described method of purification of alpha galactosides, members of the raffinose family of oligosaccharides (RFOs), has been developed for lupins. The considerable amount of sucrose present in the RFO preparations obtained by the previous method has been removed by modifying the purification stage on diatomaceous earth and charcoal. The present method allows for the preparation of high-purity RFOs containing approximately 99.4% of RFOs in the form of a white fine powder, which provides new perspectives for the production of pure alpha-galactoside preparations for their use as prebiotics in functional foods. PMID- 15537298 TI - Identification of angiotensin-I-converting enzyme inhibitory peptides derived from sodium caseinate hydrolysates produced by Lactobacillus helveticus NCC 2765. AB - Angiotensin-I-converting enzyme (ACE) inhibitory activity was identified in milk proteins fermented with Lactobacillus (Lb.) helveticus NCC 2765 (Nestle Culture Collection, Vers-chez-les-Blanc, Switzerland). Hydrolyzing sodium caseinate for 1 and 2 h inhibited ACE activity, as measured by an in vitro ACE inhibition test. The hydrolysates with the highest ACE inhibitory potential were fractionated by gel permeation chromatography and their low molecular weight fractions collected. These fractions were subsequently subfractionated by reverse-phase high-pressure liquid chromatography. Several hydrophobic subfractions showed high ACE inhibitory potential, and their peptide composition was determined using an ion trap mass spectrometer equipped with an elctrospray ionization source. Analysis of the low molecular weight fraction identified 14 peptides with known antihypertensive activity and 1 with previously described opioid activity. On the basis of the peptide composition of active subfractions, two potentially active novel sequences were defined, and the following synthetic peptides were synthesized: FVAPFPEVFG (alphaS1 39-48), ENLLRFFVAPFPEVFG (alphaS1 33-48), NENLLRFFVAPFPEVFG (alphaS1 32-48), LNENLLRFFVAPFPEVFG (alphaS1 31-48), NLHLPLPLL (beta 147-155), ENLHLPLPLL (beta 146-155), and VENLHLPLPLL (beta 145-155). The ACE inhibitory potential of these synthetic peptides was assessed, and IC50 values were determined. NLHLPLPLL (beta 147-155), which was the only synthetic peptide also present in the sodium caseinate hydrolysates, and NENLLRFFVAPFPEVFG (alphaS1 32-48) showed the highest inhibition of ACE activity, with IC50 values of 15 and 55 microM, respectively. Furthermore, the stability of all synthetic peptides was assessed using an in vitro model simulating gastric digestion. The beta-casein-derived peptides remained intact following the successive hydrolysis by pepsin and pancreatin, whereas alphaS1-casein-derived peptides were degraded by pepsin. PMID- 15537299 TI - Evaluation of the antimicrobial activity of citrus essences on Saccharomyces cerevisiae. AB - The aim of this research was to assess the antimicrobial activity of nine different industrial essences used in a soft drink factory in relation to their composition, as well as to verify the role of vapor pressure on their bioactivity. The essences were tested against a Saccharomyces cerevisiae strain isolated from spoiled soft drinks. The tests were carried out by adding the essences directly to a liquid medium or into the headspace of closed systems inoculated with the yeast. The headspace composition was evaluated through a solid phase microextraction-gas chromatography technique. The use of a mass spectrometer allowed the identification of the peaks detected. The microbial growth was indirectly monitored by measuring the metabolic CO2 released by the yeast. The results obtained indicated that the most effective essences were characterized by the highest concentration of some terpenes, such as citral, beta pinene, and p-cymene. Moreover, all of the essences were more bioactive when added directly to the liquid medium. PMID- 15537300 TI - Tumor cell proliferation and cyclooxygenase enzyme inhibitory compounds in Amaranthus tricolor. AB - Amaranthus tricolor is consumed as a vegetable in Asia. Bioassay-directed isolation of leaves and stems of A. tricolor yielded three galactosyl diacylglycerols (1-3) with potent cyclooxygenase and human tumor cell growth inhibitory activities. The purified compounds were characterized by spectroscopic methods. In addition, the fatty acid moieties in diacyl galactosyl glyerols were characterized by GC-MS analyses. The galactosyl diacylglycerols 1-3 inhibited the cyclooxygenase-1 (COX-1) enzyme by 78, 63, and 93% and the cyclooxygenase-2 (COX 2) enzyme by 87, 74, and 95%, respectively. These compounds were tested for antiproliferative activity using human AGS (gastric), CNS (central nervous system; SF-268), HCT-116 (colon), NCI-H460 (lung), and MCF-7 (breast) cancer cell lines. Compound 1 inhibited the growth of AGS, SF-268, HCT-116, NCI-H460, and MCF 7 tumor cell lines with IC50 values of 49.1, 71.8, 42.8, 62.5, and 39.2 mug/mL, respectively. For AGS, HCT-116, and MCF-7 tumor cell lines, the IC50 values of compounds 2 and 3 were 74.3, 71.3, and 58.7 microg/mL and 83.4, 73.1, and 85.4, respectively. This is the first report of the COX enzyme inhibitory activity for galactosyl glycerols and antiproliferative activities against human colon, breast, lung, stomach, and CNS tumor cell lines. PMID- 15537301 TI - Insect juvenile hormone analogues and their biological activity on sea lice (Lepeophtheirus salmonis). AB - A number of synthetic compounds that exhibit juvenile hormone activity when tested on the insect species Tenebrio molitor have been shown to inhibit the development of the sea lice species Lepeophtheirus salmonis. The testing was carried out on both isolated nauplius larvae and live Atlantic salmon (Salmo salar). PMID- 15537302 TI - Correlation of glucosinolate content to myrosinase activity in horseradish (Armoracia rusticana). AB - Fully developed horseradish (Armoracia rusticana Gaertn., Mey., & Scherb.) roots from 27 accessions and leaves from a subset of 9 accessions were evaluated for glucosinolates and myrosinase enzyme activity. Eight different glucosinolates were detected (based on HPLC retention times as desulfoglucosinolates) in both root and leaf tissues. The sum of these glucosinolates, referred to as total, ranged from 2 to 296 micromol g(-1) of dry weight (DW) in both tissues. Four glucosinolates (sinigrin, glucobrassicin, neoglucobrassicin, and gluconasturtiin) were detected in major quantities. In fully developed roots, sinigrin concentration represented approximately 83%, gluconasturtiin approximately 11%, and glucobrassicin approximately 1% of the total glucosinolates. Approximately the same proportions of individual glucosinolates appeared in fully developed leaves, except that glucobrassicin was substituted by neoglucobrassicin and gluconasturtiin concentration was significantly lower (<1%). At least four other glucosinolates were detected in very small quantities (<1%) in both roots and leaves. Myrosinase (beta-thioglucoside glucohydrolase, EC 3.2.3.1) is the enzyme responsible for the hydrolysis of the parent glucosinolates into biologically active products. Very little is known about myrosinase activity and the correlation of its activity to total and individual glucosinolates in plant tissues. Significant differences in myrosinase activity were detected between the roots and leaves, ranging from 1.2 to 57.1 units g(-1) of DW. Data showed no correlation between myrosinase activity and total and/or individual glucosinolates in the roots. However, in the leaves, significant correlations were found between myrosinase activity and total glucosinolates (0.78 at P = 0.01) and between myrosinase activity and sinigrin (0.80 at P = 0.01). Glucosinolates content and myrosinase activity were also correlated in young and fully developed roots and leaves and during tissue crushing. Glucobrassicin concentration in the roots and neoglucobrassicin concentration in the leaves were significantly higher in young than in fully developed tissue. Crushing of the tissue resulted in rapid hydrolysis of sinigrin and glucobrassicin, as expected, from the presence of myrosinase. Likewise, myrosinase activity declined rapidly after crushing, perhaps due to inactivation by the reaction products and/or the depletion of its substrates. PMID- 15537303 TI - Greek plant extracts exhibit selective estrogen receptor modulator (SERM)-like properties. AB - To prevent bone loss that occurs with increasing age, nutritional and pharmacological factors are needed. Traditional therapeutic agents (selective estrogen receptor modulators or SERMs, biphosphonates, calcitonin) may have serious side effects or contraindications. In an attempt to find food components potentially acting as SERMs, we submitted four plant aqueous extracts derived from Greek flora (Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum) in a series of in vitro biological assays reflective of SERM profile. We examined their ability (a) to stimulate the differentiation and mineralization of osteoblastic cell culture by histochemical staining for alkaline phosphatase and Alizarin Red-S staining, (b) to induce, like antiestrogens, the insulin growth factor binding protein 3 (IGFBP3) in MCF-7 breast cancer cells, and (c) to proliferate cervical adenocarcinoma (HeLa) cells by use of MTT assay. Our data reveal that all the plant extracts studied at a concentration range 10-100 microg/mL stimulate osteoblastic cell differentiation and exhibit antiestrogenic effect on breast cancer cells without proliferative effects on cervical adenocarcinoma cells. The presence of estradiol inhibited the antiestrogenic effect induced by the extracts on MCF-7 cells, suggesting an estrogen receptor-related mechanism. In conclusion, the aqueous extracts derived from Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum may form the basis to design "functional foods" for the prevention of osteoporosis. PMID- 15537304 TI - Qualitative and quantitative evaluation of the genomic DNA extracted from GMO and non-GMO foodstuffs with four different extraction methods. AB - The presence of DNA in foodstuffs derived from or containing genetically modified organisms (GMO) is the basic requirement for labeling of GMO foods in Council Directive 2001/18/CE (Off. J. Eur. Communities 2001, L1 06/2). In this work, four different methods for DNA extraction were evaluated and compared. To rank the different methods, the quality and quantity of DNA extracted from standards, containing known percentages of GMO material and from different food products, were considered. The food products analyzed derived from both soybean and maize and were chosen on the basis of the mechanical, technological, and chemical treatment they had been subjected to during processing. Degree of DNA degradation at various stages of food production was evaluated through the amplification of different DNA fragments belonging to the endogenous genes of both maize and soybean. Genomic DNA was extracted from Roundup Ready soybean and maize MON810 standard flours, according to four different methods, and quantified by real-time Polymerase Chain Reaction (PCR), with the aim of determining the influence of the extraction methods on the DNA quantification through real-time PCR. PMID- 15537306 TI - Modified mutation method for screening low citrinin-producing strains of Monascus purpureus on rice culture. AB - Monascus purpureus NTU 601 is a strain that produces monacolin K, gamma aminobutyric acid (GABA), and citrinin under solid culture conditions. Because citrinin is a mycotoxin and possesses nephrotoxic and hepatoxic effects, it has a negative impact on the acceptance of red mold rice by people. In this research, a simple and quick selection method for mutant strains with low citrinin production was designed based on the fact that citrinin possesses antibacterial activity for Bacillus subtilis and will form an inhibition zone around the colony of the Monascus strain. The mutant strain M. purpureus N 301 only produced 0.23 +/- 0.01 ppm citrinin, which was 50% less than that of the parent strain, and the monacolin K production was 481.29 +/- 7.98 ppm and maintained 91% productivity. M. purpureus N 310, the other mutant strain, produced 0.27 +/- 0.01 ppm citrinin, which was 41% less than that of the parent strain, and the monacolin K production was 526.29 +/- 5.54 ppm, which showed no significant changes when compared with the parent strain. The GABA content of the two strains was 5000 ppm, which is similar to that of the parent strain. The results showed that the method could be used to select red mold rice with low citrinin production. PMID- 15537305 TI - Bollgard II cotton: compositional analysis and feeding studies of cottonseed from insect-protected cotton (Gossypium hirsutum L.) producing the Cry1Ac and Cry2Ab2 proteins. AB - Bollgard II cotton event 15985 producing the Cry1Ac and Cry2Ab2 proteins has been developed by genetic modification to broaden the spectrum of insects to which the plant is tolerant and to provide an insect resistance management tool to impede the onset of resistance. The purpose of this study was to evaluate the composition and nutrition of Bollgard II cotton, relative to the use for food and animal feed, compared to that of conventional cotton varieties. Compositional analyses were conducted to measure proximate, fiber, amino acid, fatty acid, gossypol, and mineral contents of cottonseed from a total of 14 U.S. field sites over two years. Compositional analysis results showed that the cottonseed and cottonseed oil from Bollgard II cotton were comparable in their composition to those of the conventional control cotton line and other commercial varieties. The composition data are supported by nutritional safety studies conducted with dairy cows, catfish, and quail. Results from these studies showed that Bollgard II performed similarly to the conventional control cotton varieties. These data demonstrate that Bollgard II cotton is compositionally and nutritionally equivalent to conventional cotton varieties. These data support the conclusion that Bollgard II cotton is as safe and nutritious as conventional cotton for food and feed use. PMID- 15537308 TI - Production of soy protein concentrates using a combination of electroacidification and ultrafiltration. AB - Soy protein concentrates produced by combining electroacidification and dead-end ultrafiltration with a membrane of 100 kDa (pH 7 and 6) were compared with concentrates produced by ultrafiltration (pH 9) and a traditional acid precipitation procedure at pH 4.5. Mineral removal during ultrafiltration (mainly potassium, phosphorus, and calcium) was enhanced for the pH 6 electroacidified extract, compared to the extract at pH 9. This yielded a concentrate with improved solubility characteristics. The solubility for the concentrate prepared at pH 6 was enhanced by as high as 45% when compared to the concentrate at pH 9. The concentrate produced according to the traditional acid precipitation process showed mineral contents and solubility profile similar to those of the pH 6 concentrate, but required twice as much water during the process. The effect of electroacidification treatments on ultrafiltration permeate flux was quantified through the measurement of the different hydraulic resistances. Cake resistance was the main resistance to the permeate flux, and it was minimum at pH 9, maximum at pH 7, and intermediate at pH 6. PMID- 15537309 TI - Effect of gas environment and sorbate addition on flavor characteristics of irradiated apple cider during storage. AB - Apple cider, with (0.1%) and without potassium sorbate, was packaged in polystyrene containers and exposed to three different gas environments: oxygen flush, nitrogen flush, and atmospheric air. To evaluate the effects of irradiation (2 kGy) and storage on flavor and microbial quality, these irradiated apple cider samples were compared to a control, unirradiated sample exposed to atmospheric air. Volatile compounds, soluble solids, titratable acidity, and microbiological counts were determined weekly throughout 7 weeks of refrigerated (4 degrees C) storage. Cider irradiated and stored in atmospheric air or nitrogen flush environments had lower rates of loss for characteristic flavor volatiles compared to unirradiated apple cider and cider irradiated and stored in an oxygen flush environment. The addition of potassium sorbate to the apple cider resulted in lower counts of yeasts and aerobic microorganisms, reduced fermentation of sugars to organic acids, and improved retention of volatile compounds characteristic of apple cider. PMID- 15537307 TI - Enzymatic production and complete nuclear magnetic resonance assignment of the sugar lactulose. AB - The enzymatic transgalactosylation from lactose to fructose leading to the prebiotic disaccharide lactulose was investigated using the beta-galactosidase from Aspergillus oryzae and the hyperthermostable beta-glycosidase from Pyrococcus furiosus (CelB). The conditions for highest lactulose yields relative to the initial lactose concentration were established on a 1 mL scale. Dependent on the initial molar ratio of lactose to fructose, more or fewer oligosaccharides other than lactulose were generated. Bioconversions on a 30 mL scale in a stirred glass reactor were performed, and lactulose yields of 46 mmol/L (44% relative to lactose) for CelB and 30 mmol/L (30% relative to lactose) for A. oryzae beta galactosidase were achieved. Only <5% of other oligosaccharides were detectable. The corresponding productivities were 24 and 16 mmol/L/h, respectively. The molecular structure of lactulose was investigated in detail and confirmed after purification of the reaction solution by LC-MS and 1D and 2D NMR. Lactulose (4-O beta-D-galactopyranosyl-D-fructose) was unambiguously proved to be the major transglycosylation disaccharide. PMID- 15537310 TI - Changes in contents of carotenoids and vitamin E during tomato processing. AB - The aim of this study was to investigate the effect of different types of tomato processing on contents of lycopene, beta-carotene, and alpha-tocopherol. Samples of tomato sauce, tomato soup, baked tomato slices, and tomato juice were taken at different times of heating, respectively, after each step of production. HPLC was used to analyze contents of carotenoids and vitamin E. Due to the loss of water during thermal processing, contents of lycopene, beta-carotene, and alpha tocopherol on a wet weight basis increased. On a dry weight basis, contents of lycopene increased or decreased depending on the origin of the tomatoes used, whereas the beta-carotene contents decreased or were quite stable. In contrast to lycopene, beta-carotene isomerized due to thermal processing. The alpha tocopherol contents significantly rose during short-term heating. The increase was not caused by release of alpha-tocopherol from the seeds containing predominantly gamma-tocopherol and accounting for 2% of total alpha-tocopherol content only. PMID- 15537311 TI - Effective ways of decreasing acrylamide content in potato crisps during processing. AB - The aim of this work was to examine the effect of blanching or soaking in different acid solutions on the acrylamide content in potato crisps. Furthermore, the effects of a shorter frying time and a lower frying temperature combined with a postdrying were investigated. Soaking or blanching of potato slices in acidic solutions decreased the pH of potato juice and increased the extraction of amino acids and sugars. Potato crisps obtained after such pretreatments were characterized by lower acrylamide content. The most effective extraction of free amino acids and sugars as well as the largest decrease of acrylamide content (90%) in crisps was obtained when potato slices were soaked in acetic acid solution for 60 min at 20 degrees C. Shorter frying time followed by postdrying resulted in low-moisture potato crisps. Furthermore, the postdrying treatment gave a decreases in acrylamide content of approximately 70% when potato slices were fried at 185 degrees C and approximately 80% when potato slices were fried at 160 degrees C. Effective ways of decreasing acrylamide content in crisps production have been found. Crisps with low acrylamide content and good sensory quality can be obtained either by blanching in acetic acid as pretreatment or by a short frying followed by postdrying. PMID- 15537312 TI - Effect of different washing procedures on phenolic metabolism of shredded, packaged iceberg lettuce during storage. AB - Different washing treatments applying chlorinated, ozonated, and tap water were examined for their effect on the phenolic metabolism of minimally processed iceberg lettuce (Lactuca sativa L.) during storage in consumer-sized bags at 4 degrees C for up to 9 days. To eliminate problems associated with raw material inhomogeneity, processing was conducted on a pilot-plant scale under operating conditions of industrial practice. Inherent product heterogeneity caused by diverse lettuce leaf tissues was compensated for by pooling large-sized samples, and frequent sampling ensured significant data about the activities of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), and peroxidase (POD), as well as the contents of caffeic acid derivatives over storage time. In the homogeneous lettuce samples, specific responses caused by different washing procedures were detectable. PAL activity in the samples increased for up to 5-8 days of storage. Compared to tap and ozonated water, the use of chlorinated water (100-200 mg/L free chlorine) for washing trimmed heads or shredded lettuce significantly reduced PAL activity and the concomitant rise of 3,5-di-O caffeoylquinic acid (isochlorogenic acid isomer) concentrations. The phenolic acids O-caffeoyltartaric (caftaric acid), di-O-caffeoyltartaric (chicoric acid), 5-O-caffeoylquinic (chlorogenic acid isomer), and O-caffeoylmalic were less influenced by different washing treatments. Individual contents either were constant or decreased during storage. Additionally, the novel finding of a further caffeic acid isomer, tentatively identified as meso-di-O-caffeoyltartaric acid, is reported. PPO and POD activities were less affected by different washing treatments and thus were less suitable physiological indicators of stress reactions triggered by alternative processing. PMID- 15537313 TI - Qualitative and semiquantitative analysis of phenolic compounds in extra virgin olive oils as a function of the ripening degree of olive fruits by different analytical techniques. AB - Capillary electrophoresis (CE) can be effectively used as a fast screening tool to obtain qualitative and semiquantitative information about simple and complex phenolic compounds of extra virgin olive oil. Three simple phenols (tyrosol, hydroxytyrosol, and vanillic acid), a secoiridoid derivative (deacetoxy oleuropein aglycon), and two lignans (pinoresinol and acetoxypinoresinol) were detected as the main compounds in extra virgin olive oils by high-performance liquid chromatography (HPLC) and capillary zone electrophoresis (CZE). Spectrophotometric indices, radical scavenging activity, and oxidative stability of extra virgin olive oil samples obtained from olives hand-picked at different ripening degrees were statistically correlated with the CZE and HPLC quantification. The concentration of phenols in extra virgin olive oil decreased with ripeness of olive fruits. The high correlations found between CZE and the other analytical results indicate that CE can be applied as a rapid and reliable tool to routinely determine phenolic compounds in extra virgin olive oils. PMID- 15537314 TI - Improving the fatty acid profile of fairy shrimp, Streptocephalus dichotomus, using a lipid emulsion rich in highly unsaturated fatty acids. AB - Fatty acids are the largest component of lipids and have become a useful tool in the determination of live feeds to a variety of cultured species. Bioencapsulation is a technique which allows high-level incorporation of desired components (i.e., fatty acids, vitamins, antibiotics, etc.) in live feeds, which in turn can be supplemented to the consumer organisms. The procedure described in the present study serves as a platform of technology for enriching the Streptocephalus dichotomus. Uptake of two enrichment diets (ALGAMAC2000 and DHA SELCO) by adult S. dichotomus was investigated. The fatty acid profile supports the hypothesis that the enrichment diet increases the level of essential fatty acids, such as linolic, linolenic, eicosapentenoic, and docosahexaenoic acids. The average content (percent of total fatty acids detected) of the enriched organism by different highly unsaturated fatty acid (HUFA) products were as follows: ALGAMAC2000 showed 14-22% saturated fatty acid (SFA), 17-18% monounsaturated fatty acid (MUFA), 28-41% polyunsaturated fatty acid (PUFA), 23 34% n-3, and 4.9-7.5% n-6, whereas DHA-SELCO showed about 20-23% SFA, 20-26% MUFA, 38% PUFA, 28-31% n-3, and 7.5-10% n-6. Our present investigation proves that both HUFA-rich diets appear to be an appropriate enrichment diet, and further provides an additional rationale for using fairy shrimp as a maturation diet for any cultivable freshwater organism. PMID- 15537315 TI - Comparison of anthocyanin pigment and other phenolic compounds of Vaccinium membranaceum and Vaccinium ovatum native to the Pacific Northwest of North America. AB - Two huckleberry species, Vaccinium membranaceum and Vaccinium ovatum, native to Pacific Northwestern North America, were evaluated for their total, and individual, anthocyanin and polyphenolic compositions. Vaccinium ovatum had greater total anthocyanin (ACY), total phenolics (TP), oxygen radical absorbing capacity (ORAC), and ferric reducing antioxidant potential (FRAP) than did V. membranaceum. The pH and degrees Brix were also higher in V. ovatum. Berry extracts from each species were separated into three different fractions- anthocyanin, polyphenolic, and sugar/acid-by solid-phase extraction. The anthocyanin fractions of each species had the highest amount of ACY, TP, and antioxidant activity. Each species contained 15 anthocyanins (galactoside, glucoside, and arabinoside of delphinidin, cyanidin, petunidin, peonidin, and malvidin) but in different proportions. Their anthocyanin profiles were similar by high-performance liquid chromatography with photodiode array detection (LC DAD) and high-performance liquid chromatography with photodiode array and mass spectrometry detections (LC-DAD-MS). Each species had a different polyphenolic profile. The polyphenolics of both species were mainly composed of cinnamic acid derivatives and flavonol glycosides. The major polyphenolic compound in V. membranaceum was neochlorogenic acid, and in V. ovatum, chlorogenic acid. PMID- 15537317 TI - Nucleophilic radical substitution reaction of triazine herbicides with polysulfides. AB - Triazine herbicides are among the most widely used herbicides in the United States. Many triazine compounds are relatively stable under natural conditions and have become prominent contaminants in hydrologic systems. It was previously reported that chloro-s-triazine compounds were rapidly dechlorinated in water by polysulfides, and the reaction was assumed to be aromatic nucleophilic substitution (SNAr). In this study, we evaluated the effect of free radical inhibitors on the reaction rate of polysulfides with herbicides atrazine, simazine, and cyanazine. The reaction was significantly inhibited by radical scavengers oxygen and 1,4-benzoquinone, suggesting involvement of free radicals in the reaction. Spectral analysis of the reaction mixture using electron spin resonance showed that after the reaction, the free radical concentration in polysulfide solution substantially decreased. These evidences indicate that radical sulfur anions may also be involved in the reaction, likely via a free radical substitution reaction (SRN1) mechanism. Amendment of sodium tetrasulfide significantly reduced the leaching of atrazine or simazine from packed sand columns. Therefore, polysulfide salts may be potentially used to remove residues of triazine herbicides in environmental media. PMID- 15537316 TI - Levels of organochlorine pesticides in soils and rye plant tissues in a field study. AB - The organochlorine pesticides are lipophilic and persistent and tend to accumulate in soils and growing plants. The contamination of growing plants occurs by adhesion of volatile substances from the air to the plant surface and by the migration of contaminants through xylem in inner ascendant transport. Persistent organochlorine pesticides (HCB, alpha,gamma-HCH, pp'DDE, op'DDT, pp'DDT) levels were determined in soils and rye plants. The aims of the study were the monitoring of organochlorine pesticide concentrations and the comparison of these levels among soil, rye straw, and rye grains. Fifty soil samples and 50 rye plant (50 straw and 50 grains) samples were taken. The GLC-ECD chromatographic results indicated the following contamination levels distributed among soil, straw, and grains: HCB (0.7-1.2-0.7 microg.kg(-1)), alpha-HCH (0.6 3.4-1.2 microg.kg(-1)), gamma-HCH (1.8-27.3-4.4 microg.kg(-1)), Sigma-HCH (2.5 30.7-5.6 microg.kg(-1)), pp'DDE (1.0-7.8-5.5 microg.kg(-1)), op'DDT (16.1-20.4 17.0 microg.kg(-1)), pp'DDT (38.0-41.7-49.6 microg.kg(-1)), and Sigma-DDT (54.2 63.2-72.1 microg.kg(-1)). The study verified the presence of organochlorine pesticides in the Mexican agricultural environment and their migration from soil to the growing rye plants. However, DDT has been banned since 1999 for sanitary reasons, and Lindane is applied only in some cases as a seed dresser. The determined organochlorine pesticide levels in rye plants are low, at residual levels that are below Codex Alimentarius Commission maximum residue limits. PMID- 15537318 TI - Foliar and soil deposition of pesticide sprays in peanuts and their washoff and runoff under simulated worst-case rainfall conditions. AB - There are few studies that relate the timing and amounts of pesticide washoff from plant foliage during rainfall to runoff losses at the edge of the field. We hypothesized that foliar deposits, if washed onto the soil slowly during rainfall, may then undergo less leaching during the period of infiltration that occurs prior to soil saturation and runoff, thus exhibiting larger runoff losses than pesticides on/in the soil at the beginning of rain. We measured the runoff of ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT dye using simulated rainfall on 450 m2 mesoplots planted in peanut. Ethalfluralin was applied preplant incorporated, and metolachlor was applied preemergence on bare soil. Chlorothalonil and rhodamine WT were applied to the peanut canopy at maturity. Rainfall was simulated 24 h after each chemical application (in May and July, 1998, and May and August, 1999) using raindrop sprinklers, applying 5.5 +/- 0.5 cm over a 2 h period to create reasonable worst-case conditions; between 3 and 9 mm of runoff was generated. Volume-weighted average concentrations of chemicals in runoff were 7, 104, 163, and 179 ug L(-1) for ethalfluralin, metolachlor, chlorothalonil, and rhodamine WT, respectively. The total amounts of chemicals lost in the runoff events were 0.04 +/- 0.01, 0.2 +/- 0.1, 0.6 +/- 0.5, and 0.2 +/- 0.1, as percents of amounts applied, respectively. Rhodamine WT formed a vivid red solution on wetting and provided visual clues to the dynamics of chemical washoff/runoff. The washoff from rain-exposed peanut foliage appeared to be complete within a few minutes of the beginning of rainfall, and disappearance of dye from rain-exposed soil surface occurred within the first 10 min of rainfall. However, dye was present in runoff water at near-constant concentrations throughout the 2 h runoff event, indicating that near-constant amounts of chemical remained in the soil extraction zone. These results confirm earlier studies showing that soil incorporation at application significantly reduces runoff losses and that a majority of foliar residues can be washable if rainfall occurs within a few days after application. Runoff losses of foliar applied pesticides were small relative to washoff amounts but were sensitive to runoff timing relative to washoff. PMID- 15537319 TI - Aroma volatility from aqueous sucrose solutions at low and subzero temperatures. AB - The gas-liquid partition coefficients of ethyl acetate and ethyl hexanoate have been measured in water and aqueous sucrose solutions from 25 to -10 degrees C by dynamic headspace. Experiments were carried out on sucrose solutions at temperatures where no ice formation was possible. Results showed that when sucrose concentration increased, aroma volatility increased except for ethyl hexanoate and in the highest sucrose concentration solution (57.5%). A quasi linear temperature decrease on aroma volatility was observed in sucrose solutions from 25 to around 4 and 0 degrees C. Then, from 0 to -10 degrees C, aroma volatility did not decrease: ethyl acetate volatility remained constant but that of ethyl hexanoate increased. Enthalpy of vaporization and activity coefficients of the aroma compounds were calculated. PMID- 15537320 TI - Changes of astringent sensation of soy milk during tofu curd formation. AB - The effect of isoflavone on soy milk and tofu astringency was investigated, and no consistency was found between an undesirable astringent taste and isoflavone contents. Isoflavone-enriched extract (approximately 39% isoflavones) showed no astringency. Soybean foods having high amounts of isoflavones showed less astringency. About 80% of isoflavones exist freely in both soy milk and tofu, but 55% of phytates (which play an important role in the formation of the tofu curd network) exist freely in the soy milk, and 6-13%, on the basis of coagulation, existed freely in the tofu curds. A 1% potassium phytate solution at pH 7 showed the very same astringency as soy milk; however, calcium phytate at the same concentration and pH showed no undesirable sensation. Thus, it is assumed that the astringent characteristics caused by phytic ions in soy milk are lost upon conversion of phytic ions to their insoluble salt forms during soy milk coagulation. PMID- 15537321 TI - Synthesis of deuterated gamma-lactones for use in stable isotope dilution assays. AB - Two syntheses of deuterated gamma-lactones for use as internal standards in stable isotope dilution assays (SIDA) were developed. [2,2,3,3-2H4]-gamma-Octa-, gamma-deca-, and -gamma-dodecalactones with >89% deuterium incorporation were prepared in 27, 17, and 19% overall yields, respectively, by the reduction of a doubly protected hydroxypropiolic acid with deuterium gas. [3,3,4-2H3]-gamma-Octa and -gamma-dodecalactones were prepared in 6 and 23% yields with >92% deuterium incorporation by the free radical addition of 2-iodoacetamide to [1,1,2-2H3]-1 hexene and [1,1,2-2H3]-1-decene, respectively. Reaction yields were highly dependent upon the purity of the 1-alkene starting material. The deuterated gamma lactones were evaluated as internal standards for SIDA. PMID- 15537322 TI - Contribution of dimethyl sulfide to the aroma of Syrah and Grenache Noir wines and estimation of its potential in grapes of these varieties. AB - The contribution of dimethyl sulfide (DMS) to the aroma of Syrah and Grenache Noir wines from the Rhone Valley of France was investigated by sensory analysis, and its levels in these wines were measured. The potential DMS in the corresponding grapes and wines, susceptible to release during wine aging, was evaluated. Free DMS and potential DMS assessed by a heat-alkaline treatment were measured in grape juices and wines by SPME-GC-MS using methods previously reported and slightly modified. A relationship between potential DMS from grapes and the total DMS levels in wine was demonstrated. Furthermore, a linear regression between the ratio of free DMS levels to these total DMS levels in wine and time of storage was found. Free and potential DMS levels in grapes and wines depended on grape variety, vintage, and vine location. DMS imparted a noticeable and complex contribution to the aroma of the wines investigated, depending on the mode of sensory perception used, either before or after glass swirling. It significantly enhanced the fruity notes of the wines, and additional truffle and black olive notes. PMID- 15537323 TI - Effects of enzymatic deamidation by protein-glutaminase on structure and functional properties of alpha-zein. AB - The performance of novel protein-glutaminase (PG) purified from Chryseobacterium proteolyticumon alpha-zein was investigated. Highly insoluble alpha-zein was able to be deamidated to the extent of deamidation degree 62% by using 50 mM potassium phosphate (pH 8) containing 11.7% ethanol, at 40 degrees C for 137 h. Analysis by sodium dodecyl sulfate polyacrylamide-gel electrophoresis showed that deamidated and non-deamidated zeins have different mobilities. Results of circular dichroism spectra revealed the decline in alpha-helix contents of alpha-zein by deamidation. Besides, Fourier transform infrared spectroscopy analysis demonstrated alterations in the secondary structure of alpha-zein by deamidation. The assignment of the amide I region showed a remarkable decrease in antiparallel intermolecular beta-sheets (around 1690 cm(-1)) as an indication of the weakening aggregation ability of the deamidated molecules. Solubility and emulsification properties of alpha-zein, particularly at pH 7, were remarkably improved after the deamidation by PG. Gas chromatography and peroxide value studies pointed out that deamidated alpha-zein in powder form exhibited an inferior antioxidative property as compared with the non-deamidated one. PMID- 15537324 TI - Two-dimensional gel electrophoresis detection of protein oxidation in fresh and tainted rainbow trout muscle. AB - Protein oxidation is evaluated in rainbow trout muscle by labeling protein carbonyls with 2,4-dinitrophenyl hydrazine (DNPH) followed by immunoblotting of proteins separated by SDS-PAGE or two-dimensional gel electrophoresis (2D-GE). The carbonylation level is accessed on proteins in a whole muscle homogenate or proteins soluble in a high-salt or low-salt buffer. Spoilage-related changes in carbonylation are followed in the high-salt-protein and low-salt-protein fractions by 2D immunoblotting, which reveals increases regarding total number and intensity of carbonylation in both protein fractions for fish kept at room temperature for 48 h. The major amount of carbonylated proteins is found among the high-salt-soluble proteins, and this protein fraction is also responsible for the biggest increase in carbonylation during fish tainting. The results give an estimate of the level of protein carbonylation in rainbow trout and reveal that oxidation increases for a distinct number of proteins during tainting. PMID- 15537325 TI - Structure, chemical composition, and xylanase degradation of external layers isolated from developing wheat grain. AB - The external layers of wheat grain were investigated during maturation with respect to chemical and structural features and xylanase degradability. Cytochemical changes were observed in the isolated peripheral tissues of the wheat grain at four defined stages following anthesis. Marked chemical changes were highlighted at 11 days after anthesis, for which protein and lipid contents varied weakly. The profile of esterified ferulic acid showed large variation in the maturing peripheral layers of grain in contrast to the deposition of ferulate dimers, p-coumaric and sinapic acids. Lignin was monitored at the latest stages of ripening, which corresponds to the cessation of reserve accumulation in the grain. Arabinoxylans (AX) reached a maximum at 20 days and did not display any significant change in arabinosyl substitution proportion until ripeness. When submitted to xylanase, all outer layers were similarly altered in the proportion of soluble AX except for the peripheral tissues of the 11-day-aged wheat grain that had very little AX. Aleurone and nucellar layers were mostly degraded, whereas pericarp stayed intact at all stages of maturation. This degradation pattern was connected with the preferential immunolocalization of xylanase in aleurone and nucellar layers irrespective of the developmental stages. Further chemical examination of the enzyme-digested peripheral tissues of the grain supports the facts that ferulic ester is not a limiting factor in enzyme efficiency. Arabinose branching, ferulic dimers, and ether-linked monomers that are deposited early in the external layers would have more relevance to the in situ degradability of AX. PMID- 15537326 TI - Investigations on the high molecular weight foaming fractions of espresso coffee. AB - The target of the present work was the chemical, technological, and sensorial characterization of the brown polymers (foaming fractions) of freshly prepared espresso coffee. The total foaming fraction (TFF) was precipitated with ammonium sulfate from the defatted freshly prepared beverage and then subfractionated by adding 2-propanol/water to give an insoluble fraction (foaming fraction A, FFA) and a soluble fraction (foaming fraction B, FFB). The former is almost colorless, has a higher molecular weight and a lower nitrogen content, and contains mostly polysaccharides, whereas the latter has a lower molecular weight and a higher protein/melanoidin content, which results in a darker color. FFB showed greater foaming capability, but FFA contributed to the stability of the foam. FFB was further fractionated with solid-phase extraction and characterized by different analytical methods (size exclusion chromatography, UV, HPLC-DAD, 1H NMR). All of the melanoidin-rich fractions showed antioxidant properties with the 2,2-diphenyl 1-picrylhydrazyl hydrate method. PMID- 15537327 TI - Strecker-type degradation produced by the lipid oxidation products 4,5-epoxy-2 alkenals. AB - Strecker degradation is one of the most important reactions leading to final aroma compounds in the Maillard reaction. In an attempt to clarify whether lipid oxidation products may be contributing to the Strecker degradation of amino acids, this study analyzes the reaction of 4,5-epoxy-2-alkenals with phenylalanine. In addition to N-substituted 2-(1-hydroxyalkyl)pyrroles and N substituted pyrroles, which are major products of the reaction, the formation of both the Strecker aldehyde phenylacetaldehyde and 2-alkylpyridines was also observed. The aldehyde, which was produced at 37 degrees C-as could be determined by forming its corresponding thiazolidine with cysteamine-and pH 6-7, was not produced when the amino acid was esterified. This aldehyde is suggested to be produced through imine formation, which is then decarboxylated and hydrolyzed. This reaction also produces a hydroxyl amino derivative, which is the origin of the 2-alkylpyridines identified. All these data indicate that Strecker-type degradation of amino acids is produced at 37 degrees C by some lipid oxidation products. This is a new proof of the interrelations between lipid oxidation and Maillard reaction, which are able to produce common products by analogue mechanisms. PMID- 15537328 TI - Kinetic studies of cholesterol oxidation as inhibited by stearylamine during heating. AB - The formation of cholesterol oxidation products (COPs) during heating in the presence of stearylamine at 140 degrees C was analyzed by high-performance liquid chromatography (HPLC) and kinetically studied by use of nonlinear regression models. Results indicated that the COPs concentration increased with increasing heating time, and stearylamine was shown to reduce both oxidation and degradation rates of cholesterol. Without stearylamine, the highest rate constant (per hour) was observed for epoxidation (545.4), followed by free radical chain reaction (251.0), reduction (147.3), dehydration (95.8), triol dehydrogenation (4.7), degradation (0.34), triol formation (0.31), and dehydrogenation (0.13). With stearylamine, the epoxidation and free radical chain reaction rates could be reduced by about 800- and 3.4-fold, respectively, and triol formation during oxidation could be completely inhibited. In addition, the reactions for reduction, dehydration, degradation, and dehydrogenation could proceed slower in the presence of stearylamine. The kinetic model developed in this study can be used to predict the inhibition of COPs formation by stearylamine during heating of cholesterol. PMID- 15537329 TI - Effect of selenate supplementation on glycoalkaloid content of potato (Solanum tuberosum L.). AB - Potatoes (Solanum tuberosum L.) supplemented with increasing amounts of sodium selenate were analyzed for glycoalkaloid (GA) content. GAs were extracted with 5% acetic acid from freeze-dried tubers of two potato cultivars, Satu and Sini, harvested 10 weeks after planting as immature. The GAs alpha-solanine and alpha chaconine were quantified by reverse-phase high-performance liquid chromatography (RP-HPLC) with diode array detection. Two independent experiments were performed. In the first experiment, the total GA concentration +/- standard error of the tubers ranged between 105 +/- 9 and 124 +/- 10 mg kg(-1) fresh weight in Satu and between 194 +/- 26 and 228 +/- 10 mg kg(-1) fresh weight in Sini. The ratio of alpha-solanine to alpha-chaconine was 0.2 in Satu and 0.5-0.6 in Sini. In the second experiment, the total GA concentration +/- standard error was 75 +/- 4 to 96 +/- 11 mg kg(-1) fresh weight, and the ratio of alpha-solanine to alpha chaconine was 0.3-0.4 in Satu. A high sodium selenate supplementation (0.9 mg of Se kg(-1) quartz sand) slightly decreased the GA content in Satu, but this decrease was not statistically significant. Furthermore, at this addition level the Se concentration increased to a very high level of 20 microg g(-1) dry weight, which cannot be recommended for human consumption. In both experiments, the Se concentration in tubers increased with increasing sodium selenate application levels. Our results show that acceptable application levels of selenate did not have an effect on the GA concentration in immature potato tubers. PMID- 15537330 TI - Mass spectrometric evidence for the existence of oligomeric anthocyanins in grape skins. AB - The fractionation of a grape skin extract by multilayer countercurrent chromatography coupled with step gradient elution allowed the preparation of a fraction almost devoid of free anthocyanins. This fraction appeared to be almost exclusively polymeric, as judged by liquid chromatographic-mass spectrometric (LC MS) analysis, color-bleaching tests with sulfur dioxide, and thiolysis. Electrospray mass spectrometric analysis indicated that the pigmented material in this fraction was chiefly composed of direct condensation products of anthocyanin extending up to trimers. With regard to their linkages, the anthocyanin units in the oligomers were possibly linked by either an A-type (by both carbon-carbon and ether bonds) or B-type (by carbon-carbon bond) linkage, like proanthocyanidins. The terminal anthocyanin unit of the oligomers is consistently in the flavylium form but the extension units are in the flavan form for the A-type oligomers and in the flavene form for the B-type oligomers. Although their linkages still need to be defined rigorously, this is the first mass spectrometric evidence confirming the existence of anthocyanin oligomers in the grape skin extract. PMID- 15537331 TI - Potential chemopreventive properties of extract from baked sweet potato (Ipomoea batatas Lam. Cv. Koganesengan). AB - The extract from baked sweet potato (Ipomoea batatas Lam. cv. Koganesengan) showed potential cancer-preventing effects. The extract was partially fractionated to four fractions (I, II-a II-b, and III) by Sephadex G-25 gel chromatography. The cytotoxicity against human myelocytic leukemia HL-60 cells, the suppression of TPA-induced transformation in mouse skin JB6 C141 cells, the apoptosis inducing activity in HL-60 cells, and the scavenging capacity against DPPH radical were tested on the four fractions. Fractions II-a and III showed markedly strong radical scavenging effects on the DPPH radical, coinciding with the high content of total phenolic compounds in the fractions. Both of these fractions suppressed strongly the proliferation of HL-60 cells with apoptosis induction in a dose-dependent manner. Moreover, the two fractions markedly blocked TPA-induced cell transformation in the JB6 cell line. Taken together, these data suggest that the water extract from baked sweet potato had potential chemopreventive properties. PMID- 15537332 TI - Effect of flavonoids on stress responses in myotube cultures. AB - Effects of flavonoids on stress response of myotube cultures was studied by monitoring the release of [14C] taurine, leukotriene production, and 2',7' dichlorodihydroflourescein (DCFH2) oxidation. Stress was induced by hypotonic shock, which was accompanied by cell swelling leading to increased leukotriene production and a concomitant increase in reactive oxygen species and osmolyte release. In this model system, addition of the flavonoids catechin and quercetin decreased leukotriene production, DCFH2 oxidation, and taurine efflux, indicating a reduction of cellular stress. High concentrations of epigallocatechin gallate (EGCG) and tea extract increased leukotriene production and initial DCFH2 oxidation, indicating an increased cellular stress (possibly toxicity). However, taurine efflux was reduced, and also longer exposure time as well as lower concentrations of EGCG and tea reduced DCFH2 oxidation. Trolox and alpha tocopherol did not significantly affect taurine efflux or leukotriene production, and it was therefore concluded that suppression of these responses was not confined to redox activity in a myotube culture. PMID- 15537333 TI - Characterization of the antioxidant activity of sugars and polyhydric alcohols in fish oil emulsions. AB - Polyols have been incorporated into fish oil emulsions as a means for the inhibition of lipid oxidation and suppression of fishy flavor. However, the role of sugars and polyhydric alcohols as antioxidants has not been clearly established. Selected polyols were evaluated for their performance as antioxidants and modifiers of oxidation pathways in a model system. Oil/water (O/W) emulsions were prepared with freshly steam-deodorized menhaden oil. A layer of emulsion in aluminum pans held at 5 degrees C was exposed to 2550 lx fluorescent lights for 24 h before peroxide values and volatile flavor compounds were analyzed by GC headspace entrainment procedure. Antioxidant activity was confirmed for fructose, sucrose, raffinose, sorbitol, or mannitol when incorporated at 16% of the aqueous phase into model fish oil-in-water emulsions. Peroxide values were suppressed 10-18% in treated samples compared to control samples. Viscosity data did not exclude possible contributions from a restricted oxygen diffusion mechanism in the antioxidant activity, but revealed that emulsion viscosity did not govern fish oil oxidation rates. Combining polyols with phenolic antioxidants (alpha-tocopherol, BHT, or TBHQ) frequently diminished the antioxidant activity compared to that for individual phenolic antioxidants, which was interpreted as indicating that the H-donating activity of phenolic antioxidants was hindered by the H-bonding activity of polyols. A viscosity-based inhibition of the retroaldol conversion of (E,Z)-2,6-nonadienal to (Z)-4-heptenal with a high fructose concentration (67%) was attributed to a restriction of molecular mobility of reactants, but the conversion was only slightly inhibited by the concentration of fructose (16%) used in experimental emulsions. The data supported a hypothesis that either or both free radical scavenging and transition state metal chelation activities were provided by polyols in fish oil emulsions. Also, polyols retarded the water-requiring retroaldol decomposition of (E,Z)-2,6 nonadienal to (Z)-4-heptenal in the model systems and the reaction may be involved in some suppression of fishy flavors in emulsions. PMID- 15537334 TI - Uptake of quercetin and quercetin 3-glucoside from whole onion and apple peel extracts by Caco-2 cell monolayers. AB - Evidence suggests that regular consumption of fruits and vegetables may reduce the risk of chronic diseases, and phytochemicals from fruits and vegetables may be responsible for this health benefit. However, there is limited knowledge on the bioavailability of specific phytochemicals from whole fruits and vegetables. This study used Caco-2 cells to examine uptake of quercetin aglycon and quercetin 3-glucoside as purified compounds and from whole onion and apple peel extracts. Pure quercetin aglycon was absorbed by the Caco-2 cells in higher concentrations than quercetin 3-glucoside (p < 0.05). Caco-2 cells treated with quercetin 3 glucoside accumulated both quercetin 3-glucoside and quercetin. Caco-2 cells absorbed more onion quercetin aglycon than onion quercetin 3-glucoside (p < 0.05), and the percentage of onion quercetin absorbed was greater than that of pure quercetin, most likely due to enzymatic hydrolysis of quercetin 3-glucoside and other quercetin glucosides found in the onion by the Caco-2 cells. Caco-2 cells absorbed low levels of quercetin 3-glucoside from apple peel extracts, but quercetin aglycon absorption was not detected. Caco-2 cell homogenates demonstrated both lactase and glucosidase activities when incubated with lactose and quercetin 3-glucoside, respectively. This use of the Caco2 cell model appears to be a simple and useful system for studying bioavailability of whole food phytochemicals and may be used to assess differences in bioavailability between foods. PMID- 15537335 TI - Simultaneous and sensitive detection of three foodborne pathogens by multiplex PCR, capillary gel electrophoresis, and laser-induced fluorescence. AB - The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 x 10(3) cfu mL(-1) of S. aureus, 570 cfu mL(-1) of L. monocytogenes, and 790 cfu mL(-1) of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL(-1) of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way. PMID- 15537338 TI - Discovery of positive allosteric modulators for the metabotropic glutamate receptor subtype 5 from a series of N-(1,3-diphenyl-1H- pyrazol-5-yl)benzamides that potentiate receptor function in vivo. AB - This report describes the discovery of the first centrally active allosteric modulators of the metabotropic glutamate receptor subtype 5 (mGluR5). Appropriately substituted N-(1,3-diphenyl-1H-pyrazol-5-yl)benzamides (e.g., 8) have been identified as a novel class of potent positive allosteric modulators of mGluR5 that potentiate the response to glutamate. An iterative analogue library synthesis approach provided potentiators with excellent potency and selectivity for mGluR5 (vs mGluRs 1-4, 7, 8). Compound 8q demonstrated in vivo proof of concept in an animal behavior model where known antipsychotics are active, supporting the development of new antipsychotics based on the NMDA hypofunction model for schizophrenia. PMID- 15537337 TI - Piperidine-based nocaine/modafinil hybrid ligands as highly potent monoamine transporter inhibitors: efficient drug discovery by rational lead hybridization. AB - Some piperidine-based nocaine/modafinil hybrid ligands have been designed, synthesized, and found to display an improved potency at all three monoamine transporters and particularly for DAT and/or NET. Some highly active and selective monoamine transporter inhibitors with low nanomolar to subnanomolar potency were identified. Ligands of this type may find important applications as positron emission tomography imaging tools and in the treatment of central nervous system disorders such as depression and sleep apnea. PMID- 15537339 TI - An orally bioavailable, functionally selective inverse agonist at the benzodiazepine site of GABAA alpha5 receptors with cognition enhancing properties. AB - (3-tert-Butyl-7-(5-methylisoxazol-3-yl)-2-(1-methyl-1H-1,2,4-triazol-5 ylmethoxy)pyrazolo[1,5-d][1,2,4]triazine (13) has been identified as a functionally selective, inverse agonist at the benzodiazepine site of GABA(A) alpha5 receptors. 13 is orally bioavailable, readily penetrates the CNS, and enhances performance in animal models of cognition. It does not exhibit the convulsant, proconvulsant, or anxiogenic activity associated with nonselective GABA(A) inverse agonists. PMID- 15537340 TI - Novel purine nitrile derived inhibitors of the cysteine protease cathepsin K. AB - Starting from the high-throughput screening hit 1a, novel cathepsin K inhibitors have been developed based on a purine scaffold. High-resolution X-ray structures of several derivatives have revealed the binding mode of these unique cysteine protease inhibitors. PMID- 15537341 TI - Development of cobalt(3,4-diarylsalen) complexes as tumor therapeutics. AB - [1,6-Bis(2-hydroxyphenyl)-3,4-diaryl-2,5-diazahexa-1,5-diene]cobalt(II) complexes (cobalt(3,4-diarylsalen)) with 2-, 3-, or 4-OCH(3)/OH substituents in the 3,4 standing aryl rings were synthesized and tested for antitumor activity in vitro on the MCF-7, MDA-MB 231, and LNCaP/FGC cell lines. The cytotoxicity depended on both the configuration of the diene ligand and the kind of substituents in the 3,4-standing aromatic rings. d,l-7 (2-OCH(3)), d,l-8 (3-OCH(3)), and d,l-9 (4 OCH(3)) were equipotent to cisplatin, while the respective hydroxy-substituted complexes (d,l-10 (2-OH), d,l-11 (3-OH), and d,l-12 (2-OH)) as well as all of the meso-configured compounds (m-7 to m-12) did not influence the cell growth. Interestingly, a high catalytic potency and a rapid and high accumulation in MCF 7 cells (15- to 25-fold compared to the cell culture medium (5 microM)) were demonstrated for m-7 (2-OCH(3)), m-8 (3-OCH(3)), and m-9 (4-OCH(3)). Therefore, a mode of action based on a cobalt-catalyzed oxidative damage of the DNA is not very likely. PMID- 15537342 TI - A novel site-directed affinity reagent for cross-linking human hemoglobin: bis[2 (4-phosphonooxyphenoxy)carbonylethyl]phosphinic acid. AB - Bis[2-(4-phosphonooxyphenoxy)carbonylethyl]phosphinc acid (BPPCEP) was prepared and evaluated as a site-directed affinity reagent for cross-linking human hemoglobin. It was synthesized in four steps starting from 4-benzyloxyphenol and was converted to its pentasodium salt so as to afford efficient cross-linking in an aqueous medium. The reagent was found to specifically cross-link human hemoglobin A(0) in the beta-cleft chains under oxygenated reaction conditions at neutral pH. The amino acid residues involved in the cross-linking were determined by mass spectral analyses of tryptic digest fragments of cross-linked hemoglobin, employing a MALDI-TOF mass spectrometer. The MS analyses suggested that the most likely amino acids involved in the cross-links are Val-1 or Lys-82 present on one of the beta subunits and Lys-82 or Lys-144 on the other. Molecular modeling studies performed on the reagent-HbA(0) complex corroborated the conclusions reached by MALDI-MS analyses. The oxygen equilibrium measurements of the three major BPPCEP-cross-linked Hb products, isolated and purified by preparative cation exchange chromatography, exhibited oxygen affinity (P(50)) values of 14.5, 12.1, and 15.5 Torr as compared with the P(50) of 13.1 Torr for cell-free hemoglobin. The oxygen-binding cooperativity of the modified products, as determined by the Hill coefficient generated from the Hill plots of the respective P(50) values, coupled with the absence of sigmoidal shape of the O(2) equilibrium curves, was considerably lower than that of the native hemoglobin. PMID- 15537343 TI - Fluorinated phenylcyclopropylamines. 2. Effects of aromatic ring substitution and of absolute configuration on inhibition of microbial tyramine oxidase. AB - A series of para-substituted diastereopure cis- and trans-2-fluoro-2 arylcyclopropylamines were synthesized and these were investigated as inhibitors of microbial tyramine oxidase from Arthrobacter sp. All compounds were shown to be competitive inhibitors of this enzyme. The nature of the para-substituents in the more potent trans-isomer (cis-relationship between fluorine and the amino group) of 2-fluoro-2-arylcyclopropylamine influenced the inhibitory potency in a consistent fashion. Thus, electron-withdrawing groups (F, Cl) slightly decreased the activity, while the methyl group (+ I substituent) increased the activity by a factor of ca. 7 compared to trans-2-fluoro-2-phenylcyclopropylamine and by a factor of 90 compared to tranylcypromine. Activity also was strongly dependent on the absolute configuration. The (1S,2S)-enantiomer of 2-fluoro-2 phenylcyclopropylamine was an excellent inhibitor of tyramine oxidase whereas the (1R,2R)-enantiomer was essentially devoid of activity. PMID- 15537344 TI - Pyrazolo[1,5-a]pyrimidines: estrogen receptor ligands possessing estrogen receptor beta antagonist activity. AB - In our search for novel subtype-selective estrogen receptor (ER) ligands, we have examined various heterocyclic units as core structural elements. Here, we have investigated the fused, bicyclic pyrazolo[1,5-a]pyrimidine core, which is a system that allows for analogues to be readily assembled in a library-like fashion. This series of pyrazolo[1,5-a]pyrimidine ER ligands provided us with a new pharmacological profile for an ER ligand: compounds that are passive on both ERs, with a distinct potency selectivity in favor of ERbeta. The most distinctive ligand in this series, 2-phenyl-3-(4-hydroxyphenyl)-5,7-bis(trifluoromethyl) pyrazolo[1,5-a]pyrimidine, was 36-fold selective for ERbeta in binding. Curiously, on the basis of molecular modeling, the ERbeta binding selectivity of compounds in this series appears to be derived from differing orientations that they adapt in the ligand binding pockets of ERalpha vs ERbeta. In transcription assays this pyrazolopyrimidine was fully effective as an ERbeta antagonist while exhibiting no significant activity on ERalpha. Thus, this ligand functions as a potency- and efficacy-selective ERbeta antagonist that would abrogate estrogen action through ERbeta with minimal effects on its activity through ERalpha; as such, it could be used to study the biological function of ERbeta. PMID- 15537345 TI - Synthesis and biological evaluation of 1-aryl-4,5-dihydro-1H-pyrazolo[3,4 d]pyrimidin-4-one inhibitors of cyclin-dependent kinases. AB - Using a high-throughput screening strategy, a series of 1-aryl-4,5-dihydro-1H pyrazolo[3,4-d]pyrimidin-4-ones was identified that inhibit the cyclin-dependent kinase (CDK) 4/cyclin D1 complex-mediated phosphorylation of a protein substrate with IC(50)s in the low micromolar range. On the basis of preliminary structure activity relationships (SAR), a model was proposed in which these inhibitors occupy the ATP-binding site of the enzyme, forming critical hydrogen bonds to the same residue (Val96) to which the amino group in ATP is presumed to bind. X-ray diffraction studies on a later derivative bound to CDK2 support this binding mode. Iterative cycles of synthesis and screening lead to a novel series of potent, CDK2-selective 6-(arylmethyl)pyrazolopyrimidinones. Placement of a hydrogen-bond donor in the meta-position on the 6-arylmethyl group resulted in approximately 100-fold increases in CDK4 affinity, giving ligands that were equipotent inhibitors of CDK4 and CDK2. These compounds exhibit antiproliferative effects in the NCI HCT116 and other cell lines. The potency of these antiproliferative effects is enhanced in anilide derivatives and translates into tumor growth inhibition in a mouse xenograft model. PMID- 15537346 TI - Design of non-nucleoside inhibitors of HIV-1 reverse transcriptase with improved drug resistance properties. 1. AB - We have used a structure-based approach to design a novel series of non nucleoside inhibitors of HIV-1 RT (NNRTIs). Detailed analysis of a wide range of crystal structures of HIV-1 RT-NNRTI complexes together with data on drug resistance mutations has identified factors important for tight binding of inhibitors and resilience to mutations. Using this approach we have designed and synthesized a novel series of quinolone NNRTIs. Crystal structure analysis of four of these compounds in complexes with HIV-1 RT confirms the predicted binding modes. Members of this quinolone series retain high activity against the important resistance mutations in RT at Tyr181Cys and Leu100Ile. PMID- 15537347 TI - Design of non-nucleoside inhibitors of HIV-1 reverse transcriptase with improved drug resistance properties. 2. AB - HIV-1 nonnucleoside reverse transcriptase inhibitors (NNRTIs) are part of the combination therapy currently used to treat HIV infection. The features of a new NNRTI drug for HIV treatment must include selective potent activity against both wild-type virus as well as against mutant virus that have been selected by use of current antiretroviral treatment regimens. Based on analogy with known HIV-1 NNRTI inhibitors and modeling studies utilizing the X-ray crystal structure of inhibitors bound in the HIV-1 RT, a series of substituted 2-quinolones was synthesized and evaluated as HIV-1 inhibitors. PMID- 15537348 TI - Synthesis of C2-C3'N-linked macrocyclic taxoids. Novel docetaxel analogues with high tubulin activity. AB - Novel C2-C3'N-linked macrocyclic taxoids 4 bearing an aromatic ring at position C2 were synthesized. These compounds, tethered between N3' and the C2-aromatic ring at the ortho, meta, or para position, were constructed by ring-closing metathesis. The para-substituted derivatives were unable to stabilize microtubules, whereas the ortho- and meta-substituted compounds show significant activity in cold-induced microtubule disassembly assay. The meta derivative 4c is the first C2-C3'-linked cyclic analogue to be equipotent to paclitaxel in this assay and to show significant cytotoxicity. Computational studies of the conformational behavior of these compounds indicate that they can adopt several conformations including mainly the "T-shaped" forms. Docking experiments have shown that the "T-shaped" form is preferred for a good interaction of these compounds with the beta-tubulin binding pocket. PMID- 15537349 TI - Design, synthesis, and biological evaluation of conformationally restricted rivastigmine analogues. AB - Rivastigmine (1), an acetylcholinesterase (AChE) inhibitor approved in 2000 for the treatment of Alzheimer disease, bears a carbamate moiety in its structure, which is able to react covalently with the active site of the enzyme. Kinetic and structural studies on the interaction of 1 with different cholinesterases have been published, giving deeper, but not definitive, insights on the catalysis mechanism. On the basis of these findings and in connection with our previous studies on a series of benzopyrano[4,3-b]pyrrole carbamates as AChE inhibitors, we designed a series of conformationally restricted analogues of 1 by including the dimethylamino-alpha-methylbenzyl moiety in different tricyclic systems. A superimposition between the conformation of 1 and the carbon derivative 4, as obtained from Monte Carlo simulations, supported the idea that the tricyclic derivatives might act as rigid analogues of 1. The biological profile of 4-9, assessed in vitro against human AChE and BChE, validated our rational design. Compound 5, bearing a sulfur-containing system, showed the highest inhibitory activity, being 192-fold more potent than 1. In the present study, the most potent inhibitors were always methyl derivatives 3-5, endowed with a nanomolar range potency, whereas the ethyl ones were 40 times less potent. A reasonable explanation for this finding might be a steric hindrance effect between the ethyl group of 1 and His440 in the active site, as already suggested by the crystal structure of the complex AChE/1. The unfavorable influence of the carbamic N alkyl chain on AChE inhibition is less striking when considering BChE inhibition, since BChE is characterized by a bigger acyl binding pocket than AChE. In fact, methyl carbamates 3-5 did not show AChE/BChE selectivity, whereas compounds 6-9 were significantly more potent in inhibiting BChE than AChE activity. At 100 microM, 5 was found to inhibit the AChE-induced aggregation only by 19% likely because it is not able to strongly interact with the peripheral anionic site of AChE, which plays an essential role in the Abeta aggregation mediated by the enzyme but is lacking in BChE structure. PMID- 15537350 TI - Improved structure-activity relationship analysis of HIV-1 protease inhibitors using interaction kinetic data. AB - Despite the availability of large amounts of data for HIV-protease inhibitors and their effectiveness with wild type and resistant enzyme, there is limited knowledge about how this and other information can be systematically applied to the development of new antiviral compounds. To identify in vitro parameters that correlate with the efficacy of HIV inhibitors in cell culture, the relationships between inhibition, interaction kinetic, and cell culture parameters for HIV-1 protease inhibitors were analyzed. Correlation, cluster, and principal component analysis of data for 37 cyclic and linear compounds revealed that the affinities (K(D)) determined from SPR-biosensor binding studies correlated better to cell culture efficacy (ED(50)) than that of the inhibition constants (K(i)), indicating that the conventional use of K(i) values for structure-activity relationship analysis of HIV-1 inhibitors should be seriously reconsidered. The association and dissociation kinetic rate constants (k(on) and k(off)) alone showed weak correlations with ED(50) values. However, ED(50) values were most related to the free enzyme concentration in the viral particle ([E]), calculated from the rate constants and the total enzyme concentration in a viral particle. A structure-activity relationship analysis of the current data set was found to be valid for all classes of compounds analyzed. In summary, use of affinity, based on interaction kinetic rate constants, rather than inhibition constants, and theoretical consideration of the physiological conditions in the virus particle provide improved structure-activity relationship analysis of HIV-1 protease inhibitors. PMID- 15537351 TI - Tetrahydrobiopterin binding to aromatic amino acid hydroxylases. Ligand recognition and specificity. AB - The three aromatic amino acid hydroxylases (phenylalanine, tyrosine, and tryptophan hydroxylase) and nitric oxide synthase (NOS) all utilize (6R)-l erythro-5,6,7,8-tetrahydrobiopterin (BH(4)) as cofactor. The pterin binding site in the three hydroxylases is well conserved and different from the binding site in NOS. The structures of phenylalanine hydroxylase (PAH) and of NOS in complex with BH(4) are still the only crystal structures available for the reduced cofactor-enzyme complexes. We have studied the enzyme-bound and free conformations of BH(4) by NMR spectroscopy and molecular docking into the active site of the three hydroxylases, using endothelial NOS as a comparative probe. We have found that the dihydroxypropyl side chain of BH(4) adopts different conformations depending on which hydroxylase it interacts with. All the bound conformations are different from that of BH(4) free in solution at neutral pH. The different bound conformations appear to result from specific interactions with nonconserved amino acids at the BH(4) binding sites of the hydroxylases, notably the stretch 248-251 (numeration in PAH) and the residue corresponding to Ala322 in PAH, i.e., Ser in TH and Ala in TPH. On the basis of analysis of molecular interaction fields, we discuss the selectivity determinants for each hydroxylase and explain the high-affinity inhibitory effect of 7 tetrahydrobiopterin specifically for PAH. PMID- 15537352 TI - 5-substituted tetrazoles as bioisosteres of carboxylic acids. Bioisosterism and mechanistic studies on glutathione reductase inhibitors as antimalarials. AB - Plasmodium parasites are exposed to elevated fluxes of reactive oxygen species during intraerythrocytic life. The most important antioxidative systems are based on the glutathione reductases of the malarial parasite Plasmodium falciparum and the host erythrocyte. The development of menadione chemistry has led to the selection of the carboxylic acid 6-[2'-(3'-methyl)-1',4'-naphthoquinolyl] hexanoic acid M(5) as an inhibitor of the parasitic enzyme. As reported here, revisiting the mechanism of M(5) action revealed an uncompetitive inhibition type with respect to both NADPH and glutathione disulfide. Masking the polarity of the acidic function of M(5) by ester or amide bonds improved antiplasmodial activity. Bioisosteric replacement of the carboxylic function by tetrazole to increase bioavailability and to maintain comparable acidity led to improved antimalarial properties as well, but only with the cyanoethyl-protected tetrazoles. Using computed ab initio quantum methods, detailed analyses of the electronic profiles and the molecular properties evidenced the similarity of M(5) and the bioisoteric tetrazole T(4). The potential binding site of these molecules is discussed in light of the recently solved crystallographic structure of P. falciparum enzyme. PMID- 15537353 TI - Building predictive models for protein tyrosine phosphatase 1B inhibitors based on discriminating structural features by reassembling medicinal chemistry building blocks. AB - A new approach to predicting the biological activity of small molecule pharmaceutics is demonstrated. Structural features of medicinal chemistry building blocks are used as 2-D molecular descriptors. These descriptors include predefined structural features and macrostructures obtained from a supervised process in which features in the core library are reassembled to provide larger features that strongly differentiate the desired biological response variable. Chemical features derived in this manner can serve as predictor variables for diverse modeling algorithms, and application using partial least squares techniques is demonstrated here. Models are presented for inhibition by benzofuran and benzothiophene biphenyl analogues of protein tyrosine phosphatase 1B (PTP1B), a target for insulin-resistant disease states. Results are compared to models for PTP1B inhibitors available in the literature based on CoMFA-related techniques and 3-D molecular descriptors. PMID- 15537354 TI - Design, synthesis, and biological evaluation of the first selective nonpeptide AT2 receptor agonist. AB - The first druglike selective angiotensin II AT(2) receptor agonist (21) with a K(i) value of 0.4 nM for the AT(2) receptor and a K(i) > 10 microM for the AT(1) receptor is reported. Compound 21, with a bioavailability of 20-30% after oral administration and a half-life estimated to 4 h in rat, induces outgrowth of neurite cells, stimulates p42/p44(mapk), enhances in vivo duodenal alkaline secretion in Sprague-Dawley rats, and lowers the mean arterial blood pressure in anesthetized, spontaneously hypertensive rats. Thus, the peptidomimetic 21 exerts a similar biological response as the endogenous peptide angiotensin II after selective activation of the AT(2) receptor. Compound 21, derived from the prototype nonselective AT(1)/AT(2) receptor agonist L-162,313 will serve as a valuable research tool, enabling studies of the function of the AT(2) receptor in more detail. PMID- 15537355 TI - AT2-selective angiotensin II analogues containing tyrosine-functionalized 5,5 bicyclic thiazabicycloalkane dipeptide mimetics. AB - This paper reports the synthesis of two angiotensin II analogues with tyrosine functionalized 5,5-bicyclic thiazabicycloalkane dipeptide mimetics replacing the Tyr(4)-Ile(5) residues. The preparation of these analogues relies on the synthesis and incorporation of an alpha,alpha-disubstituted chimeric amino acid derivative and on-resin bicyclization to a cysteine residue. The synthesized analogues both displayed high angiotensin AT(2)/AT(1) receptor binding preferences and had AT(2) receptor affinities in the same low nanomolar range as angiotensin II itself. Conformational analysis, using experimental constraints derived from NMR studies, indicated that the Tyr(4) and His(6) residues in one of the angiotensin II analogues were in close proximity to each other. PMID- 15537356 TI - Highly potent 1-aminocyclohexane-1-carboxylic acid substituted V2 agonists of arginine vasopressin. AB - The synthesis and some pharmacological properties of two sets of analogues, one consisting of six peptides with 1-aminocyclohexane-1-carboxylic acid (Acc) in position 2 and the other with the amino acid in position 3, have been described. All the peptides were tested for their pressor, antidiuretic, and uterotonic in vitro activities. The Acc(2) modification has been shown to selectively modulate the activities of the analogues. Four of the compounds were highly potent antidiuretic agonists with different pressor and uterotonic activities. On the other hand, the 3-substituted counterparts failed to exhibit any of the activities. One exception was provided by the [Mpa(1),Acc(3),Val(4),D-Arg(8)]VP analogue, which exhibited antidiuretic activity matching that of AVP, yet, unlike AVP, it was fairly selective. PMID- 15537357 TI - Centrally acting and metabolically stable thyrotropin-releasing hormone analogues by replacement of histidine with substituted pyridinium. AB - Metabolically stable and centrally acting thyrotropin-releasing hormone (TRH) analogues were designed by replacing the central histidine with substituted pyridinium moieties. Their analeptic and acetylcholine-releasing actions were evaluated to assess their potency as central nervous system (CNS) agents. A strong experimental connection between these two CNS-mediated actions of the TRH analogues was obtained in subject animals. The analogue 3-(aminocarbonyl)-1-(3-[2 (aminocarbonyl)pyrrolidin-1-yl]-3-oxo-2-[[(5-oxopyrrolidin-2 yl)carbonyl]amino]propyl)pyridinium (1a) showed the highest (TRH-equivalent) potency and longest, dose-dependent duration of action from a series of homologous compounds in antagonizing pentobarbital-induced narcosis when administered intravenously in its CNS-permeable prodrug form (2a) obtained via reduction of the pyridinium moiety to the nonionic dihydropyridine. The maximum change in hippocampal acetylcholine concentration upon perfusion of the pyridinium-containing tripeptides into the hippocampus of rats was also achieved with 1a. No binding to the endocrine TRH receptor was measured for the TRH analogues reported here; therefore, our design afforded a novel lead for centrally acting TRH analogues. We have also demonstrated the benefits of the prodrug approach on the pharmacokinetics and brain uptake/retention of pyridinium containing TRH analogues (measured by in vivo microdialysis sampling) upon systemic administration. PMID- 15537358 TI - Beta-oxygenated analogues of the 5-HT2A serotonin receptor agonist 1-(4-bromo-2,5 dimethoxyphenyl)-2-aminopropane. AB - Activation of 5-HT(2A) serotonin receptors represents a novel approach to lowering intraocular pressure. Because 5-HT(2A) serotonin receptor agonists might also produce undesirable central effects should sufficient quantities enter the brain, attempts were made to identify 5-HT(2) serotonin receptor agonists with reduced propensity to penetrate the blood-brain barrier. 1-(4-Bromo-2,5 dimethoxyphenyl)-2-aminopropan-1-ol (6), an analogue of the 5-HT(2) serotonin receptor agonist 1-(4-bromo-2,5-dimethoxyphenyl)-2-aminopropane (DOB; 1a) bearing a benzylic hydroxyl group, was identified as a candidate structure. Of the four optical isomers of 6, the 1R,2R-isomer (6d; K(i) = 0.5 nM) was found to bind at 5 HT(2A) receptors with an affinity similar to that of R(-)DOB (K(i) = 0.2 nM). Like R(-)DOB, 6d behaved as a partial agonist (efficacy ca. 50%) in a 5-HT(2) mediated calcium mobilization assay. However, in an in vivo test of central action (i.e., stimulus generalization with rats as subjects), 6d was >15 times less potent than R(-)DOB. O-Methylation of 6d (i.e., 7d; 5-HT(2A) K(i) = 0.3 nM) resulted in an agent that behaved as a full (93% efficacy) agonist. Intraocular administration of 300 microg of 6d and 7d to ocular hypertensive monkeys was shown to reduce intraocular pressure by 20-27%. Given the route of administration (i.e., topical), and concentrations necessary to reduce intraocular pressure, compounds such as 6d should demonstrate minimal central effects at potentially useful therapeutic doses and offer useful leads for further development. PMID- 15537359 TI - Quantitative structure-activity relationship of sesquiterpene lactones as inhibitors of the transcription factor NF-kappaB. AB - Sesquiterpene lactones (SLs) are the active compounds of a variety of traditionally used medicinal plants from the Asteraceae family. They are known to possess a considerable antiinflammatory activity in different inflammation models. They inhibit the transcription factor NF-kappaB probably by alkylating cysteine38 in the DNA binding domain of the p65 subunit. Here we investigate a set of 103 different sesquiterpene lactones representing 6 structural groups (44 germacranolides, 16 heliangolides, 22 guaianolides, 9 pseudoguaianolides, 2 hypocretenolides, 10 eudesmanolides) for their NF-kappaB inhibiting properties and the resulting IC(100)-values were submitted to a QSAR study. Properties important for the inhibition potency are discussed for the whole data set and for subsets of the different structural classes. PMID- 15537360 TI - N1-substituent effects in the selective delivery of polyamine conjugates into cells containing active polyamine transporters. AB - Several N(1)-arylalkylpolyamines containing various aromatic ring systems were synthesized as their respective HCl salts. The N(1)-substituents evaluated ranged in size from N(1)-benzyl, N(1)-naphthalen-1-ylmethyl, N(1)-2-(naphthalen-1 yl)ethyl, N(1)-3-(naphthalen-1-yl)propyl, N(1)-anthracen-9-ylmethyl, N(1)-2 (anthracen-9-yl)ethyl, N(1)-3-(anthracen-9-yl)propyl, and pyren-1-ylmethyl. The polyamine architecture was also altered and ranged from diamine to triamine and tetraamine systems. Biological activities in L1210 (murine leukemia), Chinese hamster ovary (CHO), and CHO's polyamine transport-deficient mutant (CHO-MG) cell lines were investigated via IC(50) cytotoxicity determinations. K(i) values for spermidine uptake were also determined in L1210 cells. The size of the N(1) arylalkyl substituent as well as the polyamine sequence used had direct bearing on the observed cytotoxicity profiles. N(1)-Tethers longer than ethylene showed dramatic loss of selectivity for the polyamine transporter (PAT) as shown in a CHO/CHO-MG cytotoxicity screen. In summary, there are clear limits to the size of N(1)-substituents, which can be accommodated by the polyamine transporter. A direct correlation was observed between polyamine-conjugate uptake and cytotoxicity. In this regard, a cytotoxicity model was proposed, which describes a hydrophobic pocket of set dimensions adjacent to the putative PAT polyamine binding site. PMID- 15537361 TI - Design, synthesis, and preliminary pharmacological evaluation of 4 aminopiperidine derivatives as N-type calcium channel blockers active on pain and neuropathic pain. AB - Several compounds with a 4-aminopiperidine scaffold decorated on both nitrogen atoms by alkyl or acyl moieties containing the structural motifs of verapamil and of flunarizine, as well as those that are more frequent in known N-type calcium channel antagonists, have been synthesized. Antinociceptive activity on the mouse hot-plate test was used to select molecules to be submitted to further studies. Active compounds were tested in vitro on a PC12 rat pheochromocytoma clonal cell line, to evaluate their action on N-type calcium channels, and on a rat model of neuropathic pain. Two compounds that show N-type calcium channel antagonism and are endowed with potent action on pain and neuropathic pain (3 and 18) have been selected for further studies. PMID- 15537362 TI - Long hydrocarbon chain keto diols and diacids that favorably alter lipid disorders in vivo. AB - Keto-substituted hydrocarbons with 11-19 methylene and bis-terminal hydroxyl and carboxyl groups have been synthesized and evaluated in both in vivo and in vitro assays for their potential to favorably alter lipid disorders including metabolic syndrome. Compounds were assessed for their effects on the de novo incorporation of radiolabeled acetate into lipids in primary cultures of rat hepatocytes as well as for their effects on lipid and glycemic variables in obese female Zucker fatty rats [Crl:(ZUC)-faBR] following 1 and 2 weeks of oral administration. The most active compounds were found to be symmetrical with four to five methylene groups separating the central ketone functionality and the gem dimethyl or methyl/aryl substituents. Furthermore, biological activity was found to be greatest in both in vivo and in vitro assays for the tetramethyl-substituted keto diacids and diols (e.g., 10c, 10g, 14c), and the least active were shown to be the bis(arylmethyl) derivatives (e.g., 10e, 10f, 14f). Compound 14c dose dependently elevated HDL-cholesterol, reduced triglycerides, and reduced NEFA, with a minimum effective dose of 30 mg/kg/day. Compound 1 g dose-dependently modified non-HDL-cholesterol, triglycerides, and nonesterified fatty acids, with a minimum effective dose of 10 mg/kg/day. At this dose, compound 10g elevated HDL cholesterol levels 2-3 times higher than pretreatment levels, and a dose dependent reduction of fasting insulin and glucose levels was observed. PMID- 15537363 TI - Synthesis of N3,5'-cyclo-4-(beta-D-ribofuranosyl)-vic-triazolo[4,5-b]pyridin-5 one, a novel compound with anti-hepatitis C virus activity. AB - A novel anti-hepatitis C virus (HCV) agent, N(3),5'-cyclo-4-(beta-D ribofuranosyl)-vic-triazolo[4,5-b]pyridinin-5-one, was identified, and the structure was confirmed by chemical synthesis from 2-hydroxy-5-nitropyridine. PMID- 15537364 TI - Influence of molecular flexibility and polar surface area metrics on oral bioavailability in the rat. AB - The relationship of rotatable bond count (N(rot)) and polar surface area (PSA) with oral bioavailability in rats was examined for 434 Pharmacia compounds and compared with an earlier report from Veber et al. (J. Med. Chem. 2002, 45, 2615). N(rot) and PSA were calculated with QikProp or Cerius2. The resulting correlations depended on the calculation method and the therapeutic class within the data superset. These results underscore that such generalizations must be used with caution. PMID- 15537365 TI - A new class of acyclic 2-alkyl-1,1,2-triaryl (Z)-olefins as selective cyclooxygenase-2 inhibitors. AB - A new class of acyclic (Z)-2-alkyl-1,2-diphenyl-1-(4 methanesulfonylphenyl)ethenes (7) was designed for evaluation as selective cyclooxygenase-2 (COX-2) inhibitors. In vitro COX-1 and COX-2 isozyme inhibition structure-activity studies identified (Z)-1,2-diphenyl-1-(4 methanesulfonylphenyl)oct-1-ene (7d) as a potent COX-2 inhibitor (IC(50) = 0.42 microM) with a high COX-2 selectivity index (SI > 234). In a carrageenan-induced rat paw edema assay, (Z)-7d exhibited excellent antiinflammatory activity (ID(50) = 1.1 mg/kg). The molecular modeling and structure-activity data acquired indicate that (Z)-olefins having cis C-1 4-methanesulfonylphenyl and C-2 unsubstituted phenyl (or 4-acetoxyphenyl) substituents in conjunction with a C-1 phenyl ring and a C-2 alkyl substituent of appropriate length constitute a suitable template for the design of a novel class of acyclic (Z)-2-alkyl-1,1,2 triaryleth-1-ene COX-2 inhibitors. PMID- 15537366 TI - NSAID-related psychiatric adverse events: who is at risk? AB - NSAIDs are frequently used in clinical practice and they account for approximately 5-10% of all drug prescriptions. NSAID use has been associated with a risk of adverse events, which have a relevant impact on morbidity and mortality and account for a substantial increment of healthcare costs. Less common, but clinically relevant, adverse events associated with NSAID use are the impairment of the CNS and, particularly, the appearance of psychiatric symptoms. These symptoms include changes in cognition, mood state and even precipitation or exacerbation of pre-existing psychiatric disorders. This article aims to review the medical literature on published reports of NSAID-related psychiatric adverse events, identify risk factors for these events and describe mechanisms potentially involved in their onset. We identified 27 reports with data on 453 cases of NSAID-related psychiatric adverse events. Data suggest that individuals who may be particularly susceptible to such events include patients with a history of psychiatric illness and possibly parturients. Indometacin and selective cyclo-oxygenase (COX)-2 inhibitors were the most frequently reported culprit drugs; however, whether this reflects an increased incidence with these drugs compared with other NSAIDs or factors such as usage or reporting patterns is unknown.A possible explanation for the psychiatric effect of NSAIDs resides in the modulation of central neurotransmission by prostaglandins, the synthesis of which is inhibited by NSAIDs. COX-2 is a key enzyme in this process since its activity is localised in distal dendrites and dendritic spines, which are cellular specialisations involved in synaptic signalling. Dopamine is considered the most relevant neurotransmitter involved in this phenomenon. Psychiatric symptoms are a rare but relevant complication of NSAID use. This effect is probably a consequence of impairment in neurotransmission modulated by prostaglandins when NSAIDs are used by susceptible individuals. These drugs should be used with caution in high-risk individuals with pre-existing psychiatric illness, and caution may also be advisable in the postpartum period. To date, reports of NSAID-related psychiatric adverse events have most commonly involved indometacin and selective COX-2 inhibitors. Prescribers should consider warning patients of the possibility of an acute neuropsychiatric event when traditional NSAIDs or selective COX-2 inhibitors are prescribed. PMID- 15537367 TI - Practice guidelines for transdermal opioids in malignant pain. AB - Patients with moderate-to-severe malignancy-related pain require opioid pharmacotherapy. Many cancer patients continue to be prescribed subtherapeutic doses of pain medications resulting in undue suffering and diminished quality of life. Pain associated with malignancy and its treatment may exacerbate other symptoms associated with cancer, including nausea, fatigue, weakness, dyspnoea, constipation and impaired cognition. The choice of analgesic pharmacotherapy should be individualised and based on the intensity of pain reported by the patient, rather than its specific aetiology. When selecting pain management pharmacotherapy, the healthcare provider should consider the patient's pain level, activity level and any comorbid illness. Intolerable adverse effects, ineffective pain relief or a change in the patient's clinical status can dictate the need for a new pain management regimen. Healthcare providers must be able to readily quantify the relative analgesic potency when converting from one opioid to another or from one route of administration to another. Transdermal formulations of fentanyl and buprenorphine are effective pharmacotherapy that can be safely used for cancer patients with pain. However, clinicians need to be cognisant that the US/UK manufacturer's recommendations for equianalgesic dose administration of transdermal fentanyl may result in initial doses that produce subtherapeutic concentrations and unrelieved pain in some patients. A less conservative dose administration algorithm for transdermal fentanyl using a 2:1 (mg/day of oral morphine : microg/h of transdermal fentanyl) conversion ratio that considers both a review of the literature and clinical experience should help clinicians individualise cancer pain pharmacotherapy. PMID- 15537369 TI - New treatment options for erectile dysfunction in patients with diabetes mellitus. AB - Erection is a neurovascular event that involves spinal and supra spinal pathways. The final common pathway involves the release of nitric oxide (NO) from both endothelial cells and neurons, which acts as a vasodilator causing penile engorgement and erection. NO is degraded by the enzyme phosphodiesterase (PDE) type 5 in the penis. Erectile dysfunction (ED), defined as the persistent inability to achieve and/or maintain an erection sufficient for satisfactory sexual performance, results when the neurovascular pathway is interrupted by medical conditions or drugs. A 15-item self-administered questionnaire, the International Index of Erectile Function (IIEF), is one of the most useful tools to evaluate erectile function (EF) in clinical trials, although of much less use in routine clinical practice. The MMAS (Massachusetts Male Aging Study) was the first major epidemiological investigation to study the prevalence of ED. The study found that ED was three times more common in patients with diabetes mellitus. The aetiopathogenesis of ED in diabetes is multifactorial, with vascular and neural factors being equally implicated. Hyperglycaemia is believed to give rise to biochemical perturbations that lead to these microvascular changes. In the MMAS, ED in diabetes was strongly correlated with glycaemic control, duration of disease and diabetic complications. The incidence increased with increasing age, duration of diabetes and deteriorating metabolic control, and was higher in individuals with type 2 diabetes than those with type 1.ED in men with diabetes often affects their quality of life and, as patients are often reluctant to come forward with their symptoms, a carefully taken history is one of the most useful approaches in identifying affected individuals. The PDE inhibitors have revolutionised the management of ED and oral drug therapy is currently first-line therapy for the condition. These agents act by potentiating the action of intracavernosal NO, thereby leading to a more sustained erection. Sildenafil was the first PDE5 inhibitor to undergo evaluation and has been studied extensively. More recently two other agents, vardenafil and tadalafil, have been introduced. All the drugs have been shown to be effective across a wide range of aetiologies of ED, including diabetes. The drugs have been shown to improve EF domain scores, penetration and maintenance of erection, resulting in more successful intercourse. Their effects are greater at higher doses. Sildenafil and vardenafil are shorter-acting agents, while tadalafil has a longer half-life allowing the user more flexibility in sexual activity. Common adverse effects include headache, nasal congestion and dyspepsia, all actions related to inhibition of PDE5. The drugs are generally well tolerated and withdrawal from the clinical studies as a result of drug-related adverse effects were rare. The use of PDE5 inhibitors in the presence of oral nitrates is absolutely contraindicated. The clinical studies to date have not evaluated the use of one drug in the case of treatment failure with another agent. Sublingual apomorphine, which stimulates central neurogenic pathways, is a new agent and may be a suitable alternative in those patients in whom PDE5 inhibitors are ineffective or contraindicated. In clinical trials, all IIEF domains except sexual desire were found to have improved after apomorphine. The median times to erection in these studies were 18.9 and 18.8 minutes for the 2 and 3mg doses, respectively. Intraurethral and intracavernosal alprostadil may be a useful alternative when oral drug therapy is ineffective or contraindicated. The management of ED in the diabetic patient may often involve a multidisciplinary approach where psychosexual counselling and specialist urologist advice is required in addition to the skills and expertise of the diabetologist. Finally, the introduction of the new oral agents have completely revolutionised the management of ED and allowed more individuals to come forward for treatment. PMID- 15537368 TI - Recent advances in treatment strategies for atopic dermatitis. AB - A wide range of different therapeutic regimens are used for atopic dermatitis. Although many treatment modalities are well established worldwide among clinicians, only the minority of these therapy recommendations are based on results of randomised controlled trials (RCTs). To close the gap between such 'generally' recommended therapies and therapies that are based on data from controlled trials, this review focuses not only on the pharmacological and clinical aspects of the currently proven agents, but also on the advantages and disadvantages of therapies that have not yet been completely tested.A review of the available literature concerning the pharmacological profile and also the level of evidence of therapeutic efficacy of all currently known topical and systemic agents for the treatment of atopic dermatitis reveals a large gap between the knowledge concerning the pharmacological action in vitro and the evidence of clinical efficacy in many cases. We agree with the conclusion of previous reviews that numerous therapies for atopic dermatitis urgently require more independent RCTs and especially comparative trials (e.g. corticosteroids vs calcineurin inhibitors). These are required in order to facilitate the choice of therapeutic strategy for the individual treatment of atopic dermatitis, with its broad spectrum of clinical manifestations and potential complications in adult patients and, particularly, in children.Finally, we also review preclinical trials with several new drugs. Immunomodulators appear to promise a new dimension for the future of therapy for atopic dermatitis, especially for severe and otherwise refractory forms or as alternatives to corticosteroids, that is, to treat facial atopic eczema without the risk of adverse effects. PMID- 15537370 TI - Eplerenone : a review of its use in left ventricular systolic dysfunction and heart failure after acute myocardial infarction. AB - Eplerenone (Inspra) is a selective aldosterone blocker. Oral eplerenone is approved for use in patients with left ventricular (LV) systolic dysfunction and clinical evidence of heart failure following acute myocardial infarction (MI) in the US and in European countries (e.g. the UK and The Netherlands). The addition of eplerenone to standard medical therapy significantly improved mortality and morbidity in patients with LV systolic dysfunction and clinical evidence of heart failure following acute MI in the large, well designed EPHESUS (Eplerenone Post acute myocardial infarction Heart failure Efficacy and SUrvival Study) trial. The beneficial effects of eplerenone on all-cause mortality and cardiovascular mortality were seen within 30 days of randomisation. Eplerenone was generally well tolerated. Although a higher incidence of hyperkalaemia occurred with eplerenone than with placebo, the incidence of hypokalaemia was significantly lower with eplerenone treatment. Thus, the addition of eplerenone to standard medical therapy is an important new strategy for further improving mortality and morbidity in post-MI patients with LV systolic dysfunction and heart failure. PMID- 15537371 TI - Olanzapine: a review of its use in the management of bipolar I disorder. AB - Olanzapine is an atypical antipsychotic that is approved in the US and Europe for the oral treatment of acute manic episodes in patients with bipolar I disorder, and for maintenance therapy to prevent recurrence in responders. Oral olanzapine is effective in the treatment of bipolar mania, both as single agent therapy and as adjunctive therapy in combination with lithium or valproate semisodium. In the treatment of acute episodes, olanzapine is superior to placebo and at least as effective as lithium, valproate semisodium, haloperidol and risperidone in reducing the symptoms of mania and inducing remission. Additional comparative studies are required to determine the efficacy of olanzapine relative to newer atypical antipsychotics, such as quetiapine, ziprasidone and aripiprazole. Olanzapine is also effective at delaying or preventing relapse during long-term maintenance therapy in treatment responders, and is currently the only atypical antipsychotic approved for this indication. Current evidence suggests that olanzapine may be more effective than lithium in preventing relapse into mania, but not relapse into depression or relapse overall. Olanzapine is generally well tolerated, and although it is associated with a higher incidence of weight gain than most atypical agents, it has a low incidence of extrapyramidal symptoms (EPS). Therefore, oral olanzapine is a useful first-line or adjunctive agent for both the acute treatment of manic episodes and the long-term prevention of relapse into manic, depressive or mixed episodes associated with bipolar I disorder. PMID- 15537373 TI - Colorectal cancer and Coxibs. PMID- 15537374 TI - Influence of rofecoxib on experimental colonic carcinogenesis in rats. AB - AIM: To investigate the effect of a selective cyclooxigenase-2 (COX-2) inhibitor, rofecoxib, in the prevalence of experimental colon tumors in rats. EXPERIMENTAL DESIGN: Experimental study with 35 male Sprague-Dawley rats, divided into four groups: a) control group without experimental manipulation (n = 5); b) pharmacological carcinogenesis with 1-2 dimethylhydrazine dihydrocloride (n = 10); c) pharmacological carcinogenesis and addition of acetylsalicylic acid (AAS) (n = 10); and d) carcinogenesis and addition of rofecoxib (n = 10). Carcinogenesis was induced with 1-2 dimethylhydrazine at a weekly dose of 25 mg/kg for 18 weeks. Colon tumors were isolated at 20 weeks. Antiinflammatory agents were given at a dose of AAS 30 mg/kg and rofecoxib at 3 mg/kg. RESULTS: The percentage of colonic tumors was significantly reduced in the rofecoxib group. This result was found for all tumors and for the malignant lesions, adenocarcinomas. CONCLUSIONS: Rofecoxib, a selective COX-2 inhibitor, reduced the percentage of drug-induced neoplastic glandular tissue in rats. PMID- 15537375 TI - Epidemiology of chronic inflammatory bowel disease in the Northern area of Huelva. AB - OBJECTIVE: To know the different epidemiologic aspects of chronic inflammatory bowel disease (IBD) in the Northern area of the province of Huelva. MATERIAL AND METHODS: we carried out a retrospective (1980-1996) and prospective (1996-2003) study of all patients diagnosed with IBD in the Northern area of Huelva, with 77,856 inhabitants. The distribution of Crohn s disease (CD) and ulcerative colitis (UC) was analyzed, as well as sex, age and smoking habit at the time of diagnosis, familial aggregation, appendicectomy rate, phenotype (anatomical site and clinical types), extraintestinal manifestations, and immunosuppressive therapy or surgical requirements. RESULTS: 70 patients with IBD were studied, 40 with UC and 30 with CD. Sex distribution was 39 (55.7%) males (55% with CD vs 56.6% with UC; NS) and 31 (44.3%) females (45% with CD vs 43.3% with UC; NS). Mean age at presentation was 44.7 +/- 19.32 years in UC vs 32.3 +/- 16.43 in CD; p < 0.001. Familial association was 7.1%. Previous appendicectomy was referred in 2.5% of patients with UC vs 36.7% of those with CD (p < 0.001), and the percentage of smokers was also higher in CD 66.7% vs CU 12.5% (p < 0.001). Anatomical site in UC was: proctitis, 20%; proctosigmoiditis/left colon, 42.5%; extensive colitis, 25%; and pancolitis, 12.5%; in CD was: terminal ileum, 43.3%; colon, 20%; and ileo-colon, 36.7%. The disease patterns were: inflammatory 56.7%, obstruction 26.7%, and fistulization 16.7%. Extraintestinal manifestations were diagnosed in 7.5% of patients with UC vs 16.6% patients with CD; surgery was performed in one patient with UC vs 10 with CD, and one patient with UC required immunosuppressive treatment vs 12 with CD. A multivariant analysis showed that younger age and smoking habit were risk factors for CD vs advanced age in UC, in which case, the smoking habit was a protective factor. The mean incidence rate of IBD starting from 1996 and expressed in cases/100,000 inhabitants/year was 5.2 for UC and 6.6 for CD. CONCLUSIONS: The mean incidence of UC in our area was 5.2 cases/100,000/inhabitants/year, and 6.6 for CD. Patients presenting with CD are diagnosed at a significantly younger age that those with UC; the smoking habit is a risk factor for CD while it protects from UC. The characteristics of IBD in our geographical area do not differ substantially from those in other regions of Spain. PMID- 15537376 TI - Research training during medical residency (MIR). Satisfaction questionnaire. AB - INTRODUCTION: It is during Medical Residency Training (MIR) that knowledge, abilities and habits are acquired, which will shape professional activity in the future. It is therefore very likely that residents who do not acquire the necessary habits and knowledge for research activities will eventually not carry out these activities in the future. The aim of this study was to analyze the level of satisfaction of residents with his or her scientific and research training, and to determine any deficiencies with respect to this training. MATERIALS AND METHODS: The aim of the questionnaire used was to determine the level of satisfaction of residents regarding their scientific and research training during their residency period. Questionnaires were usually distributed via internal mail to all residents (MIR physicians) registered at a third level teaching hospital, with a completion rate of 78% (n = 178). RESULTS: As far as the evaluation of scientific training is concerned, 68% of residents were dissatisfied or very dissatisfied. With respect to scientific studies carried out, 49% of residents had not taken part in any, but the number of studies carried out increases as the residency progresses. On the other hand, 22% of residents reported not having started their doctoral thesis, 50% having attended doctorate courses, 24% having a title for their thesis, and only 4% having written a thesis. Doctorate courses, thesis topics, and written theses increase with the year of residency, and a greater activity may be seen in this respect in surgical departments. If we analyze help available to residents for their carrying out scientific activities, 55% reported that only selected assistant doctors would offer help, and 21% reported that no doctors would offer help. Dissatisfaction with research training increases with the year of residency. With regard to main specialist fields, it can be seen that residents in surgical fields carry out more theses, whereas central fields report less facilities. Finally, if we evaluate the influence that these variables may have on the general satisfaction of residents with his or her residency, these variables are seen to be significant factors of dissatisfaction. CONCLUSIONS: Most residents are dissatisfied with their scientific training and have relatively few facilities for developing such skills, which in turn results in a scarce number of scientific studies and doctoral theses. PMID- 15537377 TI - Early onset of organ failure is the best predictor of mortality in acute pancreatitis. AB - BACKGROUND: APACHE II is a multifactorial scoring system for predicting severity in acute pancreatitis (AP). Organ failure (OF) has been correlated with mortality in AP. OBJECTIVE: To evaluate the usefulness of APACHE II as an early predictor of severity in AP, its correlation with OF, and the relevance of an early establishment of OF during the course of AP. PATIENTS AND METHODS: From January 1999 to November 2001, 447 consecutive cases of AP were studied. APACHE II scores and Atlanta criteria were used for defining severity and OF. RESULTS: Twenty-five percent of patients had severe acute pancreatitis (SAP). APACHE II at 24 h after admission showed a sensitivity, specificity, and positive and negative predictive value of 52, 77, 46, and 84%, respectively, for predicting severity. Mortality for SAP was 20.5%. Seventy percent of patients who developed OF did so within the first 24 hours of admission, and their mortality was 52%. Mortality was statistically significant (p< 0.01) if OF was established within the first 24 hours after admission. CONCLUSIONS: APACHE II is not reliable for predicting outcome within the first 24 hours after admission and should therefore be used together with other methods. OF mostly develops within the first days after admission, if ever. The time of onset of OF is the most accurate and reliable method for predicting death risk in AP. PMID- 15537378 TI - Epidemiology and diagnosis of the pancreatic cancer. PMID- 15537379 TI - Imatinib and gastrointestinal stromal tumor (GIST): a selective targeted therapy. AB - Gastrointestinal stromal tumors are the most frequent mesenchymal tumors in the gastrointestinal tract. They originate from the interstitial cells of Cajal and are characterized by an anomalous receptor for a growth factor with tyrosine kinase activity (c-kit). This anomaly causes a permanent activation of the receptor and uncontrolled cell growth. These tumors show a poor response to traditional chemotherapy drugs, and are thus associated with low survival in cases of advanced disease. Imatinib, a tyrosine kinase inhibitor, is an example of selective targeted oncologic therapy that induces improved survival in these patients. We discuss two cases of metastatic gastrointestinal stromal tumors with a good response to imatinib, and also review the pathophysiology and treatment related outcome of this type of tumors. We include results from clinical phase III studies. PMID- 15537380 TI - Hypertransaminasemia in poorly-controlled type-1 diabetes mellitus. PMID- 15537381 TI - [Preoperative platelet count and colorectal cancer recurrence]. PMID- 15537382 TI - [Propafenone hepatotoxicity]. PMID- 15537383 TI - The membrane-anchored serine protease, TMPRSS2, activates PAR-2 in prostate cancer cells. AB - TMPRSS2 is a type II transmembrane-bound serine protease that has gained interest owing to its highly localized expression in the prostate and its overexpression in neoplastic prostate epithelium. Once activated, the serine protease domain of TMPRSS2 is released from the cell surface into the extracellular space. PAR (protease-activated receptor)-2 belongs to a family of G-protein-coupled receptors (PAR-1-4) that are activated by specific serine proteases, which are expressed in many normal and malignant cell types. Previous in vitro studies on prostate cancer cells suggest a role for PAR-2 in prostate cancer metastasis. A polyclonal anti-human TMPRSS2 antibody was generated against the TMPRSS2 serine protease domain. The antibody showed specific reactivity with recombinant expressed TMPRSS2, and so was used to extract and purify the cleaved active TMPRSS2 protease from prostate cancer cells. Reverse transcriptase PCR and Western blot analysis were used to show the expression of both TMPRSS2 and PAR-2 in the androgen-dependent LNCaP prostate cancer cell line. Treatment of LNCaP cells with the cellular immunopurified TMPRSS2 protease induced a transient increase in intracellular calcium, which is indicative of G-protein-coupled receptor activation. This calcium mobilization was inhibited by cellular pre treatment with a specific PAR-2 antagonist, but not with a PAR-1 antagonist; inhibition of the protease activity also failed to mobilize calcium, suggesting that TMPRSS2 is capable of cleaving and thereby activating the PAR-2 receptor. The calcium mobilization was also inhibited by cellular pre-treatment with suramin or 2-APB (2-aminoethoxydiphenyl borate), indicating that a G-protein pathway is involved and that subsequent calcium release is mainly from intracellular stores. The present study describes how TMPRSS2 may contribute to prostate tumour metastasis via the activation of PAR-2. PMID- 15537384 TI - Critical role for Ets, AP-1 and GATA-like transcription factors in regulating mouse Toll-like receptor 4 (Tlr4) gene expression. AB - TLR4 (Toll-like receptor 4) is essential for sensing the endotoxin of Gram negative bacteria. Mutations or deletion of the TLR4 gene in humans or mice have been associated with altered predisposition to or outcome of Gram-negative sepsis. In the present work, we studied the expression and regulation of the Tlr4 gene of mouse. In vivo, TLR4 levels were higher in macrophages compared with B, T or natural killer cells. High basal TLR4 promoter activity was observed in RAW 264.7, J774 and P388D1 macrophages transfected with a TLR4 promoter reporter vector. Analysis of truncated and mutated promoter constructs identified several positive [two Ets (E twenty-six) and one AP-1 (activator protein-1) sites] and negative (a GATA-like site and an octamer site) regulatory elements within 350 bp upstream of the transcriptional start site. The myeloid and B-cell-specific transcription factor PU.1 bound to the proximal Ets site. In contrast, none among PU.1, Ets-1, Ets-2 and Elk-1, but possibly one member of the ESE (epithelium specific Ets) subfamily of Ets transcription factors, bound to the distal Ets site, which was indispensable for Tlr4 gene transcription. Endotoxin did not affect macrophage TLR4 promoter activity, but it decreased TLR4 steady-state mRNA levels by increasing the turnover of TLR4 transcripts. TLR4 expression was modestly altered by other pro- and anti-inflammatory stimuli, except for PMA plus ionomycin which strongly increased promoter activity and TLR4 mRNA levels. The mouse and human TLR4 genes were highly conserved. Yet, notable differences exist with respect to the elements implicated in gene regulation, which may account for species differences in terms of tissue expression and modulation by microbial and inflammatory stimuli. PMID- 15537385 TI - Inhibition sites in F1-ATPase from bovine heart mitochondria. AB - High-resolution crystallographic studies of a number of inhibited forms of bovine F1-ATPase have identified four independent types of inhibitory site: the catalytic site, the aurovertin B-binding site, the efrapeptin-binding site and the site to which the natural inhibitor protein IF1 binds. Hitherto, the binding sites for other inhibitors, such as polyphenolic phytochemicals, non-peptidyl lipophilic cations and amphiphilic peptides, have remained undefined. By employing multiple inhibition analysis, we have identified the binding sites for these compounds. Several of them bind to the known inhibitory sites. The amphiphilic peptides melittin and synthetic analogues of the mitochondrial import pre-sequence of yeast cytochrome oxidase subunit IV appear to mimic the natural inhibitor protein, and the polyphenolic phytochemical inhibitors resveratrol and piceatannol compete for the aurovertin B-binding site (or sites). The non peptidyl lipophilic cation rhodamine 6G acts at a separate unidentified site, indicating that there are at least five inhibitory sites in the F1-ATPase. Each of the above inhibitors has significantly different activity against the bacterial Bacillus PS3 alpha3beta3gamma subcomplex compared with that observed with bovine F1-ATPase. IF1 does not inhibit the bacterial enzyme, even in the absence of the epsilon-subunit. An understanding of these inhibitors may enable rational development of therapeutic agents to act as novel antibiotics against bacterial ATP synthases or for the treatment of several disorders linked to the regulation of the ATP synthase, including ischaemia-reperfusion injury and some cancers. PMID- 15537386 TI - Molecular basis of N-acetylglucosaminyltransferase I deficiency in Arabidopsis thaliana plants lacking complex N-glycans. AB - GnTI (N-acetylglucosaminyltransferase I) is a Golgi-resident enzyme essential for the processing of high-mannose to hybrid and complex N-glycans. The Arabidopsis thaliana cgl mutant lacks GnTI activity and as a consequence accumulates oligomannosidic structures. Molecular cloning of cgl GnTI cDNA revealed a point mutation, which causes a critical amino acid substitution (Asp144-->Asn), thereby creating an additional N-glycosylation site. Heterologous expression of cgl GnTI in insect cells confirmed its lack of activity and the use of the N-glycosylation site. Remarkably, introduction of the Asp144-->Asn mutation into rabbit GnTI, which does not result in the formation of a new N-glycosylation site, led to a protein with strongly reduced, but still detectable enzymic activity. Expression of Asn144 rabbit GnTI in cgl plants could partially restore complex N-glycan formation. These results indicate that the complete deficiency of GnTI activity in cgl plants is mainly due to the additional N-glycan, which appears to interfere with the proper folding of the enzyme. PMID- 15537387 TI - An eleven nucleotide section of the 3'-untranslated region is required for perinuclear localization of rat metallothionein-1 mRNA. AB - Localization of mRNAs provides a novel mechanism for synthesis of proteins close to their site of function. MT1 (metallothionein-1) is a small, metal-binding protein that is largely cytoplasmic but which can be found in the nucleus. The localization of rat MT1 requires the perinuclear localization of its mRNA by a mechanism dependent on the 3'-UTR (3'-untranslated region). The present study investigates the nature of this mRNA localization signal using Chinese-hamster ovary cells transfected with gene constructs in which either MT1 or the globin coding region is linked to different sequences from the MT1 3'-UTR. Deletion, mutagenesis and antisense oligonucleotide approaches indicate that nt 45-76 of the 3'-UTR, in particular nt 66-76, are required for the localization of either MT1 mRNA or chimaeric transcripts in which a beta-globin coding region is linked to sequences from the MT1 3'-UTR. This section of the 3'-UTR contains a CACC repeat. Two mutations that are predicted to alter the secondary structure of this region also impair localization. Our hypothesis is that the perinuclear localization signal in MT1 mRNA is formed by a combination of the CACC repeat and its structural context. PMID- 15537388 TI - The prototype gamma-2 herpesvirus nucleocytoplasmic shuttling protein, ORF 57, transports viral RNA through the cellular mRNA export pathway. AB - HVS (herpesvirus saimiri) is the prototype gamma-2 herpesvirus. This is a subfamily of herpesviruses gaining importance since the identification of the first human gamma-2 herpesvirus, Kaposi's sarcoma-associated herpesvirus. The HVS ORF 57 (open reading frame 57) protein is a multifunctional transregulatory protein homologous with genes identified in all classes of herpesviruses. Recent work has demonstrated that ORF 57 has the ability to bind viral RNA, shuttles between the nucleus and cytoplasm and promotes the nuclear export of viral transcripts. In the present study, we show that ORF 57 shuttles between the nucleus and cytoplasm in a CRM-1 (chromosomal region maintenance 1)-independent manner. ORF 57 interacts with the mRNA export factor REF (RNA export factor) and two other components of the exon junction complex, Y14 and Magoh. The association of ORF 57 with REF stimulates recruitment of the cellular mRNA export factor TAP (Tip-associated protein), and HVS infection triggers the relocalization of REF and TAP from the nuclear speckles to several large clumps within the cell. Using a dominant-negative form of TAP and RNA interference to deplete TAP, we show that it is essential for bulk mRNA export in mammalian cells and is required for ORF 57-mediated viral RNA export. Furthermore, we show that the disruption of TAP reduces viral replication. These results indicate that HVS utilizes ORF 57 to recruit components of the exon junction complex and subsequently TAP to promote viral RNA export through the cellular mRNA export pathway. PMID- 15537389 TI - Redox activation of mitochondrial intermembrane space Cu,Zn-superoxide dismutase. AB - The localization of Cu,Zn-superoxide dismutase in the mitochondrial intermembrane space suggests a functional relationship with superoxide anion (O2*-) released into this compartment. The present study was aimed at examining the functionality of Cu,Zn-superoxide dismutase and elucidating the molecular basis for its activation in the intermembrane space. Intact rat liver mitochondria neither scavenged nor dismutated externally generated O2*-, unless the mitochondrial outer membrane was disrupted selectively by digitonin. The activation of the intermembrane space Cu,Zn-superoxide dismutase following the disruption of mitochondrial outer membrane was largely inhibited by bacitracin, an inhibitor of protein disulphide-isomerase. Thiol alkylating agents, such as N-methylmaleimide or iodoacetamide, decreased intermembrane space Cu,Zn-superoxide dismutase activation during, but not after, disruption of the outer membrane. This inhibitory effect was overcome by exposing mitochondria to low micromolar concentrations of H2O2 before disruption of the outer membrane in the presence of the alkylating agents. Moreover, H2O2 treatment alone enabled intact mitochondria to scavenge externally generated O2*-. These findings suggest that intermembrane space Cu,Zn-superoxide dismutase is inactive in intact mitochondria and that an oxidative modification of its critical thiol groups is necessary for its activation. PMID- 15537390 TI - Domain architecture of the smooth-muscle plasma membrane: regulation by annexins. AB - Individual signalling events are processed in distinct, spatially segregated domains of the plasma membrane. In a smooth muscle, the sarcolemma is divided into domains of focal adhesions alternating with caveolae-rich zones, both harbouring a specific subset of membrane-associated proteins. Recently, we have demonstrated that the sarcolemmal lipids are similarly segregated into domains of cholesterol-rich lipid rafts and glycerophospholipid-rich non-raft regions. In the present study, we provide a detailed structural analysis of the relationship between these proteinaceous and lipid domains. We demonstrate that the segregation of plasmalemmal protein constituents is intimately linked to that of the membrane lipids. Our results imply that lipid segregation is critical for the preservation of membrane protein architecture and essential for directional translocation of proteins to the sarcolemma. We show that the membrane lipid segregation is supported by the annexin protein family in a Ca2+-dependent manner. Eukaryotic cells harbour numerous, tissue-specific subsets of annexins. By examining the significance of this variety in a smooth muscle, we demonstrate that four different annexins target membrane sites of distinct lipid composition and that each annexin requires a different [Ca2+] for its translocation to the sarcolemma. Our results suggest that the interactions of annexins with distinct plasma membrane regions promote membrane segregation and, in combination with their individual Ca2+ sensitivity, might allow a spatially confined, graded response to a multitude of extra- or intracellular stimuli. PMID- 15537391 TI - Vanishin is a novel ubiquitinylated death-effector domain protein that blocks ERK activation. AB - The ERK (extracellular-signal regulated-kinase)/MAPK (mitogen-activated protein kinase) pathway can regulate transcription, proliferation, migration and apoptosis. The small DED (death-effector domain) protein PEA-15 (phosphoprotein enriched in astrocytes-15) binds ERK and targets it to the cytoplasm. Other DED containing proteins including cFLIP and DEDD can also regulate signal transduction events and transcription in addition to apoptosis. In the present study, we report the identification of a novel DED-containing protein called Vanishin. The amino acid sequence of Vanishin is closest in similarly to PEA-15 (61% identical). Vanishin mRNA is expressed in several mouse tissues and in both mouse and human cell lines. Interestingly, Vanishin is regulated by ubiquitinylation and subsequent degradation by the 26 S proteasome. The ubiquitinylation is complex and occurs at both the internal lysine residues and the N-terminus. We further show that Vanishin binds ERK/MAPK but not the DED proteins Fas-associated death domain, caspase 8 or PEA-15. Vanishin is present in both the nucleus and Golgi on overexpression and forces increased ERK accumulation in the nucleus in the absence of ERK stimulation. Moreover, Vanishin expression inhibits ERK activation and ERK-dependent transcription in cells, but does not alter MAPK/ERK activity. Therefore Vanishin is a novel regulator of ERK that is controlled by ubiquitinylation. PMID- 15537392 TI - Regulation of adenylate cyclase type VIII splice variants by acute and chronic Gi/o-coupled receptor activation. AB - We previously reported that acute agonist activation of G(i/o)-coupled receptors inhibits adenylate cyclase (AC) type VIII activity, whereas agonist withdrawal following chronic activation of these receptors induces AC-VIII superactivation. Three splice variants of AC-VIII have been identified, which are called AC-VIII A, -B and -C (with AC-VIII-B missing the glycosylation domain and AC-VIII-C lacking most of the C1b area). We report here that AC-VIII-A and -B, but not -C, are inhibited by acute mu-opioid and dopaminergic type D2 receptor activation, indicating that the C1b area of AC-VIII has an important role in AC inhibition by G(i/o)-coupled receptor activation. On the other hand the glycosylation sites in AC-VIII did not play a role in AC-VIII regulation. Although AC-VIII-A and -C differed in their capacity to be inhibited by acute agonist exposure, agonist withdrawal after prolonged treatment led to a similar superactivation of all three splice variants, with no significant change in AC-VIII expression. AC-VIII superactivation was not affected by pre-incubation with a cell permeable cAMP analogue, indicating that the superactivation does not depend on the agonist induced reduction in cAMP levels. The superactivated AC-VIII-A, -B and -C were similarly re-inhibited by re-application of agonist (morphine or quinpirole), returning the activity to control levels. These results demonstrate marked differences in the agonist inhibition of the AC-VIII splice variants before, but not after, superactivation. PMID- 15537394 TI - Tryptase concentrations in bronchoalveolar lavage from patients with chronic eosinophilic pneumonia. AB - In order to characterize BAL (bronchoalveolar lavage) in CEP (chronic eosinophilic pneumonia) and to investigate the possible role of mast cells and tryptase in the pathogenesis of this interstitial disease, cells and tryptase levels were determined in BAL of patients with CEP and in a group of healthy controls. The results show that a statistically significant increase in tryptase concentration was found in patients with CEP compared with the healthy controls. This is the first report that shows an increase in tryptase levels in CEP and could reflect higher mast cell activation as well as larger mast cell populations in the lungs of these patients. These results strongly support the involvement of mast cells and eosinophils in the immunopathogenesis of CEP. PMID- 15537393 TI - A microbial TRP-like polycystic-kidney-disease-related ion channel gene. AB - Ion channel genes have been discovered in many microbial organisms. We have investigated a microbial TRP (transient receptor potential) ion channel gene which has most similarity to polycystic-kidney-disease-related ion channel genes. We have shown that this gene (pkd2) is essential for cellular viability, and is involved in cell growth and cell wall synthesis. Expression of this gene increases following damage to the cell wall. This fission yeast pkd2 gene, orthologues of which are found in all eukaryotic cells, appears to be a key signalling component in the regulation of cell shape and cell wall synthesis in yeast through an interaction with a Rho1-GTPase. A model for the mode of action of this Schizosaccharomyces pombe protein in a Ca2+ signalling pathway is hypothesized. PMID- 15537396 TI - Investigating the relationship between emergency departments and complementary and alternative medicine use in Australia. PMID- 15537397 TI - Strategies for improving emergency care for children: perceptions and practicalities. PMID- 15537398 TI - Snakebite in Australia: the role of the Venom Detection Kit. PMID- 15537399 TI - Political issues in emergency medicine: the United Kingdom. AB - A recent issue of this Journal published two papers on political issues facing the specialty of emergency medicine in the USA and in Canada. This paper continues the theme from a UK perspective. In recent years there has been significant change in the National Health Service, which has had (or will have) both direct and indirect consequences for the specialty of emergency medicine and the delivery of emergency care in the UK. This article describes some of the recent National Health Service initiatives and their implications for emergency medicine. PMID- 15537400 TI - Carbon Monoxide Research Group, LDS Hospital, Utah in reply to Scheinkestel et al. and Emerson: the role of hyperbaric oxygen in carbon monoxide poisoning. AB - This comprehensive response was invited by the Editor of Emergency Medicine Australasia to allow our Group from Salt Lake City, Utah to review the two articles 'Where to now with carbon monoxide poisoning?' by Scheinkestel et al. and the accompanying COMMENTARY: 'The dilemma of managing carbon monoxide poisoning' by Emerson published in the April issue of Emergency Medicine Australasia. PMID- 15537401 TI - Use and toxicity of complementary and alternative medicines among emergency department patients. AB - OBJECTIVE: To determine the prevalence and type of complementary and alternative medicines (CAM) used by ED patients, to identify patient subgroups more likely to use CAM, the conditions for which they were taken, and the prevalence of side effects and toxicity associated with CAM use. METHODS: This was an analytical cross-sectional survey of patients at a large tertiary referral ED. Consenting patients completed a specifically designed study questionnaire, in their preferred language if required. Information was collected on CAM use on the day of presentation and within the previous week and previous year. RESULTS: Four hundred and four patients were enrolled (participation rate 97.1%). Fifty (12.4%, 95% CI 9.4, 16.1) patients had taken a CAM on the day of presentation, 203 (50.2%, 95% CI 45.3, 55.2) within the previous week and 275 (68.1%, 95% CI 63.2, 72.5) within the previous year. CAM users were significantly younger, better educated and more likely to be female than non-CAM users (P < 0.01). 103 different CAM had been taken for a wide variety of reasons. Side-effects were experienced in 53 of the 1182 CAM courses taken (side-effect rate of 4.5%). One patient presented suffering specifically from CAM toxicity. CONCLUSION: Complementary and alternative medicines are used by a considerable proportion of ED patients. The potential for side-effects, toxicity and interaction with traditional medication indicates the need for consideration of CAM use in patient assessment. PMID- 15537402 TI - Agreement between bicarbonate measured on arterial and venous blood gases. AB - OBJECTIVE: This study aims to determine the extent of agreement between venous and arterial bicarbonate for a group of emergency department patients with respiratory or metabolic illness requiring blood gas analysis as part of their evaluation. METHODS: This prospective study of patients who were deemed by their treating doctor to require an arterial blood gas analysis to determine their ventilatory or acid-base status, compared bicarbonate on an arterial and a venous sample taken as close to simultaneously as possible. Data were analysed using bias (Bland-Altman) methods. Subgroup analyses were performed for the metabolic, respiratory, chronic obstructive airways disease and acidotic subgroups. RESULTS: Two hundred and forty-six patients were entered into the study; 195 with acute respiratory disease and 51 with suspected metabolic derangement. The values of bicarbonate on arterial and venous samples showed close agreement with an average difference between the samples of 1.20 mmol/L (95% limits of agreement being 2.73 to +5.13 mmol/L). Similar agreement was found for all subgroups. CONCLUSION: Venous bicarbonate estimation shows a high level of agreement with the arterial value, with acceptably narrow 95% limits of agreement. These results suggest that venous bicarbonate estimation may be an acceptable substitute for arterial measurement. PMID- 15537403 TI - Radiological imaging to improve the emergency department diagnosis of acute appendicitis. AB - OBJECTIVES: To determine the institution's current non-therapeutic (negative) appendicectomy rate; the frequency of clinical predictors for appendicitis in patients who underwent appendicectomy; and the utilization and accuracy of ultrasound scans (USS) and computed tomography (CT) in the diagnosis of appendicitis. METHODS: A retrospective chart review was conducted in an adult, metropolitan teaching hospital. Patients who presented to the ED and underwent an appendicectomy over a 12-month period were analysed. Symptoms and signs predictive of appendicitis, results of USS and CT scans if performed, and histopathology findings were abstracted from patient records. RESULTS: Two hundred and forty patients had appendicectomies, 147 (61%) were male and the median age was 25 years (range 14-78 years). The negative appendicectomy rate was 14.3% (95% CI 9.1-21.0%) and 18.3% (95% CI 11.0-26.7%) in males and females, respectively. Abdominal pain shifting to the right iliac fossa (RIF), anorexia and RIF rebound tenderness were found more frequently in patients with positive than negative appendicectomies (P < 0.05). USS and CT scans were performed in 68 (28%) and 15 (9.5%) patients, respectively. The likelihood ratio for appendicitis in patients with a normal USS or a normal CT scan was 0.83 (95% CI 0.56-1.24) and 0.08 (95% CI 0.01-0.60), respectively. There were no false positive CT scan results. CONCLUSION: Computed tomoraphy scanning should play an increasing role in the ED management of suspected appendicitis. Our negative appendicectomy rate could potentially be halved by the introduction of CT scans in the diagnostic work up of these patients. PMID- 15537404 TI - Improving emergency care for children: a model of collaboration between emergency departments. AB - OBJECTIVES: To determine whether a partnership between a tertiary paediatric hospital and a hospital with a mixed population of children and adults can improve emergency care for children. To describe the process of creating and maintaining such a partnership. METHODS: A partnership was created and various strategies employed to improve emergency paediatric care. This was evaluated by examination of demographic data, clinical audit, patient satisfaction surveys and staff satisfaction surveys. RESULTS: Paediatric attendances increased by 12% (P = 0.001), with a 33% increase in children under the age of 5 years (P = 0.001). Despite this there was a reduction of 6% in inpatient paediatric bed usage. Staff decision making and documentation improved during paediatric patient care, and parent and staff satisfaction with care of children improved. CONCLUSIONS: This initiative was successful in improving local paediatric facilities and patient care and is a new and innovative means of improving paediatric emergency care. PMID- 15537405 TI - Priapism as a presentation of T-cell acute lymphoblastic leukaemia in a child. AB - We report a case of a child who presented to the ED with priapism. This was the initial presentation of T-cell acute lymphoblastic leukaemia (ALL). To our knowledge, this is the first report of a child with T-cell ALL presenting with priapism. This case demonstrates the importance of identifying the underlying cause of priapism, as it directly impacts on both initial and ongoing management. PMID- 15537406 TI - Epistaxis. AB - Epistaxis is a frequent complaint, and may often cause great anxiety in patients and clinicians. Epistaxis results from the interaction of factors that damage the nasal mucosal lining, affect the vessel walls, or alter the coagulability of the blood, and which may be categorized into environmental, local, systemic and medication related. The knowledge of the first aid treatment of epistaxis is very poor, amongst not only the public, but also health professionals. Immediate emergency department management of epistaxis depends on prioritized assessment and treatment, including resuscitation if necessary, together with the application of relatively simple otolaryngological techniques. There is little high quality evidence regarding routine, alternative or adjunctive treatments. PMID- 15537407 TI - Specific training in clinical forensic medicine is useful to ACEM trainees. AB - OBJECTIVE: To assess the effectiveness of a 6-month rotation in forensic medicine in developing medicolegal and forensic skills for doctors training for a fellowship with the Australasian College for Emergency Medicine (ACEM). METHODS: Over a period of 6 years 12 trainees undertook a 6-month rotation with the Victorian Institute of Forensic Medicine (VIFM) as a clinical forensic registrar. These 12 trainees were surveyed and asked structured questions regarding their perception of the usefulness of the training and the impact of the rotation on their professional development. RESULTS: Twelve replies were received (100% response rate). All respondents felt that the term had a major positive influence on their ability to examine victims of physical and sexual assault, obtain forensic specimens and prepare a medicolegal report. CONCLUSIONS: A 6-month rotation in clinical forensic medicine improves ACEM trainees perceived ability to perform a number of medicolegal tasks without interfering with other areas of professional development. PMID- 15537408 TI - Recombinant factor VIIa. AB - Human coagulation factor (F) VII is a single chain protease that circulates in the blood as a weakly active zymogen at concentrations of approximately 10 nmol/L. When converted to the active 2 chain form (FVIIa), it is a powerful initiator of haemostasis. Recombinant factor VIIa (rFVIIa, eptacog alfa, NovoSeven) is a genetically engineered product that was first introduced in 1988 for the treatment of patients with haemophilia A and B with high inhibitory antibody titres to factors VIII and IX. Recent reports in the form of case studies and series, and early trial data, have suggested a role for rFVIIa across a diverse range of indications including bleeding associated with trauma, surgery, thrombocytopaenia, liver disease and oral anticoagulant toxicity. This review describes the physiology of the coagulation pathway and in particular the role of recombinant factor VIIa. It will also focus on the emerging role of rFVIIa in both trauma and non-trauma bleeding and its potential use in the ED. PMID- 15537409 TI - Consensus statement: risk of nosocomial organophosphate poisoning in emergency departments. PMID- 15537410 TI - Cross reactivity between venomous, mildly venomous, and non-venomous snake venoms with the Commonwealth Serum Laboratories Venom Detection Kit. AB - OBJECTIVE: Studies have noted the relatively common occurrence of positive urine results with the Commonwealth Serum Laboratories Venom Detection Kit (VDK) when testing patients with suspected snakebite who are not envenomed. Possible explanations have been false positive test results or subclinical envenoming. We investigated a third possibility, that there is potential for the venom (or saliva) from mildly venomous and non-venomous snakes to give a positive reading with the VDK. METHODS: Venoms/saliva from three non-venomous and seven mildly venomous snake species were tested in the laboratory with the VDK, along with control venoms from four of the five major snake genera (Brown snake, Tiger snake, Death adder and Black snake). RESULTS: Two of the venom/saliva samples, from Gould's hooded snake (Parasuta gouldii), a mildly venomous snake, and the Black-headed python (Aspidites melanocephalus), a non-venomous snake, caused a positive test for the tiger snake genus. There was also cross-reactivity between black snake venoms and the tiger snake well of the VDK. CONCLUSIONS: This study provides a further possible explanation for 'false positive' VDK results, that is venom/saliva presence or absorption from mildly or non-venomous snakes and cross reactivity with venomous snakes on VDK testing. It has implications for antivenom use should it ever be required for more severe envenoming syndromes from mildly or moderately venomous snakes, and for further research. It reinforces the practice of only using VDK testing in patients who show definite evidence of envenoming. PMID- 15537411 TI - Medical support for Adventure Racing. AB - Adventure Racing has recently become a popular sport. In the past medical and paramedical staff were frequently positioned at checkpoints throughout the course of the race. Presently there is a developing role for appropriately qualified individuals to access the course by biking, running or swimming and then providing injured racers with on-site first aid prior to their transfer to the nearest medical checkpoint. These medical staff should have not only appropriate first aid skills but also rock rescue techniques, rappelling and other basic rope work as well as be athletically fit. This article explores the new role and attempts to outline what characteristics may be required. PMID- 15537412 TI - Emergency medicine and environmental activism. AB - Clearfell, burn and sow silviculture is promoted as being the most scientific method of harvesting wet sclerophyll forests in Tasmania. However, it fails to take into account wider social and global environmental issues such as climate change and the carbon cycle. This paper describes an emergency physician's role in the continuing debate surrounding the management of Tasmania's old-growth forests. Like evidence-based medicine, the use of science in this context has its limitations, and such comparisons are discussed. PMID- 15537413 TI - Pancreatitis induced by pegylated interferon alfa-2b in a patient affected by chronic hepatitis C. AB - A middle-aged man was admitted to the ED because of nausea and vomiting, abdominal distention and fainting. A blood analysis revealed high levels of serum amylase and lipase, confirming a diagnosis of acute pancreatitis. The history showed that the patient had self-administered a single dose of pegylated interferon alfa-2b and ribavirin daily for 7 days for chronic hepatitis C. The medications were stopped and his condition gradually improved. In agreement with the literature and the Naranjo algorythm result, pegylated interferon alfa-2b is associated with acute pancreatitis. Identification of a few signs and symptoms is the first 'signal' in preventing a serious drug-induced adverse event. PMID- 15537414 TI - Bilateral theca lutein cysts: a rare cause of acute abdomen in pregnancy. AB - Theca lutein cysts (hyperreactio luteinalis) are benign cysts usually associated with molar pregnancy. We report a case of bilateral theca lutein cysts with normal intrauterine singleton pregnancy presenting as an acute abdomen requiring surgical intervention. Laparotomy revealed bilateral theca lutein cysts one of which was torted, necessitating salpingo-ovariotomy. PMID- 15537418 TI - Carbon monoxide poisoning and hyperbaric oxygen: a fairer hearing? PMID- 15537420 TI - Response to review article on acute carbon monoxide poisoning and the role of hyperbaric oxygen. PMID- 15537421 TI - Exercise-associated hyponatraemia or hypo-osmolarity? PMID- 15537423 TI - Global Arthritis Research Network (GARN): 4th World Congress on Arthritis, Montreal, Quebec, Canada, 20-22 September 2004. Abstracts. PMID- 15537425 TI - Melatonin promoted chemotaxins expression in lung epithelial cell stimulated with TNF-alpha. AB - BACKGROUND: Patients with asthma demonstrate circadian variations in the airway inflammation and lung function. Pinealectomy reduces the total inflammatory cell number in the asthmatic rat lung. We hypothesize that melatonin, a circadian rhythm regulator, may modulate the circadian inflammatory variations in asthma by stimulating the chemotaxins expression in the lung epithelial cell. METHODS: Lung epithelial cells (A549) were stimulated with melatonin in the presence or absence of TNF-alpha(100 ng/ml). RANTES (Regulated on Activation Normal T-cells Expressed and Secreted) and eotaxin expression were measured using ELISA and real-time RT PCR, eosinophil chemotactic activity (ECA) released by A549 was measured by eosinophil chemotaxis assay. RESULTS: TNF-alpha increased the expression of RANTES (307.84 +/- 33.56 versus 207.64 +/- 31.27 pg/ml of control, p = 0.025) and eotaxin (108.97 +/- 10.87 versus 54.00 +/- 5.29 pg/ml of control, p = 0.041). Melatonin(10(-10) to 10(-6)M) alone didn't change the expression of RNATES (204.97 +/- 32.56 pg/ml) and eotaxin (55.28 +/- 6.71 pg/ml). However, In the presence of TNF-alpha (100 ng/ml), melatonin promoted RANTES (410.88 +/- 52.03, 483.60 +/- 55.37, 559.92 +/- 75.70, 688.42 +/- 95.32, 766.39 +/- 101.53 pg/ml, treated with 10(-10), 10(-9), 10(-8), 10(-7),10(-6)M melatonin, respectively) and eotaxin (151.95 +/- 13.88, 238.79 +/- 16.81, 361.62 +/- 36.91, 393.66 +/- 44.89, 494.34 +/- 100.95 pg/ml, treated with 10(-10), 10(-9), 10(-8), 10(-7), 10(-6)M melatonin, respectively) expression in a dose dependent manner in A549 cells (compared with TNF-alpha alone, P < 0.05). The increased release of RANTES and eotaxin in A549 cells by above treatment were further confirmed by both real-time RT-PCR and the ECA assay. CONCLUSION: Taken together, our results suggested that melatonin might synergize with pro-inflammatory cytokines to modulate the asthma airway inflammation through promoting the expression of chemotaxins in lung epithelial cell. PMID- 15537426 TI - Locomotor activity in common spiny mice (Acomys cahirinuse): the effect of light and environmental complexity. AB - BACKGROUND: Rodents typically avoid illuminated and open areas, favoring dark or sheltered environments for activity. While previous studies focused on the effect of these environmental attributes on the level of activity, the present study tested whether the spatio-temporal structure of activity was also modified in illuminated compared with dark and complex compared with open arenas. For this, we tested common spiny mice (Acomys cahirinus) in empty or stone-containing arenas with lights on or lights off. RESULTS: In an illuminated or open arena, spiny mice moved in less frequent but longer trips with relatively long distances between consecutive stops. In contrast, in either a dark arena or an arena with stones, the animals took shorter and more frequent trips, with more stops per trip and shorter inter-stop distances. In illuminated arenas spiny mice remained mainly along the walls, whereas locomotion in the center was more prevalent in dark empty arenas, and was carried out along convoluted paths. Increasing environmental complexity by adding stones to either illuminated or dark arenas increased locomotion along straight trajectories and away from walls. CONCLUSIONS: Earlier findings of reduced activity in illuminated or open areas have been extended in the present study by demonstrating changes in the spatio temporal structure of locomotor behavior. In the more complex arenas (with stones) spiny mice traveled along short straight segments whereas in the open their trips were longer and took the shape of a zigzag path which is more effective against fast or nearby predators. Alternatively, the zigzag path may reflect a difficulty in navigation. PMID- 15537427 TI - The bag or the spindle: the cell factory at the time of systems' biology. AB - Genome programs changed our view of bacteria as cell factories, by making them amenable to systematic rational improvement. As a first step, isolated genes (including those of the metagenome), or small gene clusters are improved and expressed in a variety of hosts. New techniques derived from functional genomics (transcriptome, proteome and metabolome studies) now allow users to shift from this single-gene approach to a more integrated view of the cell, where it is more and more considered as a factory. One can expect in the near future that bacteria will be entirely reprogrammed, and perhaps even created de novo from bits and pieces, to constitute man-made cell factories. This will require exploration of the landscape made of neighbourhoods of all the genes in the cell. Present work is already paving the way for that futuristic view of bacteria in industry. PMID- 15537428 TI - Comparison of frozen and RNALater solid tissue storage methods for use in RNA expression microarrays. AB - BACKGROUND: Primary human tissues are an invaluable widely used tool for discovery of gene expression patterns which characterize disease states. Tissue processing methods remain unstandardized, leading to unanswered concerns of how to best store collected tissues and maintain reproducibility between laboratories. We subdivided uterine myometrial tissue specimens and stored split aliquots using the most common tissue processing methods (fresh, frozen, RNALater) before comparing quantitative RNA expression profiles on the Affymetrix U133 human expression array. Split samples and inclusion of duplicates within each processing group allowed us to undertake a formal genome-wide analysis comparing the magnitude of result variation contributed by sample source (different patients), processing protocol (fresh vs. frozen vs. 24 or 72 hours RNALater), and random background (duplicates). The dataset was randomly permuted to define a baseline pattern of ANOVA test statistic values against which the observed results could be interpreted. RESULTS: 14,639 of 22,283 genes were expressed in at least one sample. Patient subjects provided the greatest sources of variation in the mixed model ANOVA, with replicates and processing method the least. The magnitude of variation conferred by processing method (24 hours RNALater vs 72 hours RNALater vs. fresh vs frozen) was similar to the variability seen within replicates. Subset analysis of the test statistic according to gene functional class showed that the frequency of "outlier" ANOVA results within each functional class is overall no greater than expected by chance. CONCLUSIONS: Ambient storage of tissues for 24 or 72 hours in RNALater did not contribute any systematic shift in quantitative RNA expression results relative to the alternatives of fresh or frozen tissue. This nontoxic preservative enables decentralized tissue collection for expression array analysis without a requirement for specialized equipment. PMID- 15537429 TI - Increasing response to a postal survey of sedentary patients - a randomised controlled trial [ISRCTN45665423]. AB - BACKGROUND: A systematic review identified a range of methods, which can influence response rates. However, analysis specific to a healthcare setting, and in particular, involving people expected to be poor responders, was missing, We examined the effect of pre-warning letters on response rates to a postal survey of sedentary patients whom we expected a low rate of response. METHODS: Participants were randomised to receive a pre-warning letter or no pre-warning letter, seven days before sending the main questionnaire. The main questionnaire included a covering letter and pre-paid return envelope. After seven days, non responders were sent a reminder letter and seven days later, another reminder letter with a further copy of the questionnaire and return envelope. RESULTS: 627 adults, with a mean age of 48 years (SD 13, range 18 to 78) of whom 69.2% (434/627) were women, were randomised. 49.0% (307/627) of patients were allocated to receive a pre-warning letter and 51.0% (320/627) no pre-warning letter, seven days in advance of posting the main questionnaire. The final response rate to the main questionnaire was 30.0% (92/307) amongst those sent a pre-warning letter and 20.9% (67/320) not sent a pre-warning letter, with an adjusted odds ratio of 1.60 (95% CI 1.1, 2.30). CONCLUSIONS: The relatively low cost method of sending a pre warning letter had a modest impact on increasing response rates to a postal questionnaire sent to a group of patients for whom a low response rate was anticipated. Investigators should consider incorporating this simple intervention when conducting postal surveys, to reduce the potential for nonresponse bias and to increase the study power. Methods other than postal surveys may be needed however when a low response rate to postal surveys is likely. PMID- 15537430 TI - Uterine extracellular matrix components are altered during defective decidualization in interleukin-11 receptor alpha deficient mice. AB - BACKGROUND: Implantation of the embryo and successful pregnancy are dependent on the differentiation of endometrial stromal cells into decidual cells. Female interleukin-11 receptor alpha (IL-11Ralpha) deficient mice are infertile due to disrupted decidualization, suggesting a critical role for IL-11 and its target genes in implantation. The molecular targets of IL-11 in the uterus are unknown, but it is likely that IL-11 signaling modifies the expression of other genes important in decidualization. This study aimed to identify genes regulated by IL 11 during decidualization in mouse uterus, and to examine their expression and localization as an indication of functional significance during early pregnancy. METHODS: Decidualization was artificially induced in pseudopregnant wild type (IL11Ra+/+) and IL-11Ralpha deficient (IL11Ra-/-) littermates by oil injection into the uterine lumen, and gene expression analyzed by NIA 15K cDNA microarray analysis at subsequent time points. Quantitative real-time RT-PCR was used as an alternative mRNA quantitation method and the expression and cellular localization of the protein products was examined by immunohistochemistry. RESULTS: Among 15,247 DNA probes, 13 showed increased and 4 decreased expression in IL11Ra-/- uterus at 48 h of decidualization. These included 4 genes encoding extracellular matrix proteins; collagen III alpha1, secreted acidic cysteine-rich glycoprotein (SPARC), biglycan and nidogen-1 (entactin). Immunohistochemistry confirmed increased collagen III and biglycan protein expression in IL11Ra-/- uterus at this time. In both IL11Ra-/- and wild type uterus, collagen III and biglycan were primarily localized to the outer connective tissue and smooth muscle cells of the myometrium, with diffuse staining in the cytoplasm of decidualized stromal cells. CONCLUSION: These data suggest that IL-11 regulates changes in the uterine extracellular matrix that are necessary for decidualization. PMID- 15537431 TI - Anti-Ri antibodies associated with short-term memory deficits and a mature cystic teratoma of the ovary. AB - BACKGROUND: The IgG autoantibody ANNA-2 (anti-Ri) is a type 2 antineuronal antibody that has been found to bind to highly conserved and widely distributed adult brain proteins encoded by the Nova-1 and Nova-2 genes. Anti-Ri antibodies are typically detected in the serum and cerebrospinal fluids of patients with neurological disorders such as opsoclonus/myoclonus and cerebellar ataxia and in association with gynecologic and breast malignancies. CASE PRESENTATION: This report describes an unusual example of a 33-year-old female patient who developed short-term memory deficits over a 3-month period. An extensive neurological work up, including a panel of paraneoplastic markers was negative with the exception of a high titer serum Anti-Ri (1:15,3600). A large left ovarian mass was palpated, surgically resected and eventually diagnosed as a mature cystic teratoma. Post-operatively, memory deficits had disappeared within 1 month and serum Anti-Ri titers had decreased significantly to 1:256. An extensive diagnostic work-up for other malignancies was negative. CONCLUSION: Although, Anti-Ri antibodies are typically associated with malignancies, this case illustrates the potential association between benign tumors and this autoantibody. PMID- 15537433 TI - The emperor's new shibboleth. AB - If an idea or result can't be conveyed in plain language, how important can it really be? PMID- 15537434 TI - Management of bone loss after organ transplantation. AB - Organ transplant recipients experience rapid bone loss and high fracture rates, particularly during the early post-transplant period. Early rapid bone loss occurs in the setting of uncoupled bone turnover with increased bone resorption and decreased bone formation. Because there are no clinical factors that reliably predict post-transplant bone loss and fractures in the individual patient, all transplant recipients should be considered candidates for early preventive therapy for osteoporosis. Long-term transplant recipients with densitometric osteoporosis and/or fractures should also receive treatment. Although active metabolites of vitamin D and bisphosphonates have both shown efficacy, data from clinical trials suggest that bisphosphonates are the safest and most consistently effective agents for the prevention and treatment of post-transplantation osteoporosis in adults. Kidney transplant recipients represent a special population, and more research is needed to delineate the risks and benefits of treating bone disease in these patients. PMID- 15537435 TI - Evidence from data searches and life-table analyses for gender-related differences in absolute risk of hip fracture after Colles' or spine fracture: Colles' fracture as an early and sensitive marker of skeletal fragility in white men. AB - Based on data searches and life-table analyses, we determined the long-term (remaining lifetime) and short-term (10- and 5-year) absolute risks of hip fracture after sustaining a Colles' or spine fracture and searched for potential gender-related differences. In aging men, Colles' fractures carry a higher absolute risk for hip fracture than spinal fractures in contrast to women. These findings support the concept that forearm fracture is an early and sensitive marker of male skeletal fragility. INTRODUCTION: Colles' fracture occurrence has been largely ignored in public health approaches to identify target populations at risk for hip fracture. The aim of this study was to estimate the long-term and short-term absolute risks of hip fracture after sustaining a Colles' or spine fracture and to search for potential gender-related differences in the relationship between fracture history and future fracture risk. MATERIALS AND METHODS: To determine the long-term (remaining lifetime) and short-term (10- and 5-year) absolute risks of hip fracture, we applied life-table methods using U.S. age- and sex-specific hip fracture incidence rates, U.S. age-specific mortality rates for white women and men, pooled hazard ratios for mortality after Colles' and spine fracture, and pooled relative risks for hip fracture after Colles' and spine fracture, estimated from cohort studies by standard meta-analytic methods. RESULTS: Our results indicate that the estimated remaining lifetime risks are dependent on age in both genders. In women, remaining lifetime risks increase until the age of 80 years, when they start to decline because of the competing probabilities of fracture and death. The same pattern is found in men until the age of 85 years, the increment in lifetime risk being even more pronounced. As expected, the risk of sustaining a hip fracture was found to be higher in postmenopausal women with a previous spine fracture compared with those with a history of Colles' fracture. In men, on the other hand, the prospective association between fracture history and subsequent hip fracture risk seemed to be strongest for Colles' fracture. At the age of 50, for example, the remaining lifetime risk was 13% in women with a previous Colles' fracture compared with 15% in the context of a previous spine fracture and 9% among women of the general population. In men at the age of 50 years, the corresponding risk estimates were 8%, 6%, and 3%, respectively. Similar trends were observed when calculating 5- and 10-year risks. CONCLUSIONS: In aging men, Colles' fractures carry a higher absolute risk for hip fracture than spinal fractures in contrast to women. These findings support the concept that forearm fracture is an early and sensitive marker of male skeletal fragility. The gender-related differences reported in this analysis should be taken into account when designing screening and treatment strategies for prevention of hip fracture in men. PMID- 15537436 TI - Population-based study of age and sex differences in bone volumetric density, size, geometry, and structure at different skeletal sites. AB - In a population-based, cross-sectional study, we assessed age- and sex-specific changes in bone structure by QCT. Over life, the cross-sectional area of the vertebrae and proximal femur increased by approximately 15% in both sexes, whereas vBMD at these sites decreased by 39-55% and 34-46%, respectively, with greater decreases in women than in men. INTRODUCTION: The changes in bone structure and density with aging that lead to fragility fractures are still unclear. MATERIALS AND METHODS: In an age- and sex-stratified population sample of 373 women and 323 men (age, 20-97 years), we assessed bone geometry and volumetric BMD (vBMD) by QCT at the lumbar spine, femoral neck, distal radius, and distal tibia. RESULTS: In young adulthood, men had 35-42% larger bone areas than women (p < 0.001), consistent with their larger body size. Bone area increased equally over life in both sexes by approximately 15% (p < 0.001) at central sites and by approximately 16% and slightly more in men at peripheral sites. Decreases in trabecular vBMD began before midlife and continued throughout life (p < 0.001), whereas cortical vBMD decreases began in midlife. Average decreases in trabecular vBMD were greater in women (-55%) than in men (-46%, p < 0.001) at central sites, but were similar (-24% and -26%, respectively) at peripheral sites. With aging, cortical area decreased slightly, and the cortex was displaced outwardly by periosteal and endocortical bone remodeling. Cortical vBMD decreased over life more in women ( approximately 25%) than in men (approximately 18%, p < 0.001), consistent with menopausal-induced increases in bone turnover and bone porosity. CONCLUSIONS: Age-related changes in bone are complex. Some are beneficial to bone strength, such as periosteal apposition with outward cortical displacement. Others are deleterious, such as increased subendocortical resorption, increased cortical porosity, and, especially, large decreases in trabecular vBMD that may be the most important cause of increased skeletal fragility in the elderly. Our findings further suggest that the greater age-related decreases in trabecular and cortical vBMD and perhaps also their smaller bone size may explain, in large part, why fragility fractures are more common in elderly women than in elderly men. PMID- 15537437 TI - Primary prevention of osteoporosis: mass screening scenario or prescreening with questionnaires? An economic perspective. AB - This study focuses on the controversy surrounding selective approaches to screen for osteoporosis. Seven screening approaches were compared in terms of cost effectiveness and incremental cost-effectiveness ratios in a sample of 4035 postmenopausal women. Our results show that certain prescreening strategies are more efficient than DXA-based approaches. These results are of considerable value for health policy decision-makers and the scientific community. INTRODUCTION: There is no general consensus on the most efficient strategy to use bone densitometry for osteoporosis screening. Two distinct approaches have progressively emerged: mass screening using DXA and prescreening strategies using user-friendly risk indices. This study was designed to compare the efficiency of these approaches. MATERIALS AND METHODS: A database of 4035 medical records from postmenopausal women above 45 years was analyzed. In the first scenario, women were systematically referred to DXA if above 45, 50, or 65 years of age. The second scenario involved the validated prescreening tools SCORE, ORAI, OST, and OSIRIS and assessed two separate ways of handling their results (theoretical and pragmatic). The cost of a DXA test was set as the median Belgian value: 40.14 Euros. All strategies were compared in terms of cost exposed per osteoporotic patient detected and in terms of incremental cost-effectiveness ratios. RESULTS: In the systematic DXA strategies, the cost per patient detected ranged from 123 Euros when measuring all women >45 years of age to 91 Euros when focusing on women >65 years of age. The corresponding percentage of cases detected ranged from 100% (age > 45 years) to 50% (age > 65 years). When considering prescreening under the theoretical and pragmatic scenarios, the OSIRIS index provided the best efficiency, with costs of 74 Euros (theoretical) to 85 Euros (pragmatic) per case detected, followed by ORAI (75 Euros and 96 Euros), OST (84 Euros and 94 Euros), and SCORE (96 Euros and 103 Euros). The corresponding percentage of cases detected ranged from 89% (SCORE) to 75% (OSIRIS). The cost-effectiveness analysis showed that mass screening strategies over 50 and 65 years of age and using ORAI were best. CONCLUSIONS: Our study sets the grounds for considering, in a health economics perspective, prescreening tools as valuable, cost-effective, approaches to significantly reduce the economic burden of osteoporosis screening. PMID- 15537438 TI - Whole body BMC in pediatric Crohn disease: independent effects of altered growth, maturation, and body composition. AB - Whole body BMC was assessed in 104 children and young adults with CD and 233 healthy controls. CD was associated with significant deficits in BMC and lean mass, relative to height. Adjustment for lean mass eliminated the bone deficit in CD. Steroid exposure was associated with short stature but not bone deficits relative to height. INTRODUCTION: Children with Crohn disease (CD) have multiple risk factors for impaired bone accrual. The confounding effects of poor growth and delayed maturation limit the interpretation of prior studies of bone health in CD. The objective of this study was to assess BMC relative to growth, body composition, and maturation in CD compared with controls. MATERIALS AND METHODS: Whole body BMC and lean mass were assessed by DXA in 104 CD subjects and 233 healthy controls, 4-26 years of age. Multivariable linear regression models were developed to sequentially adjust for differences in skeletal size, pubertal maturation, and muscle mass. BMC-for-height z scores were derived to determine CD specific covariates associated with bone deficits. RESULTS: Subjects with CD had significantly lower height z score, body mass index z score, and lean mass relative to height compared with controls (all p < 0.0001). After adjustment for group differences in age, height, and race, the ratio of BMC in CD relative to controls was significantly reduced in males (0.86; 95% CI, 0.83, 0.94) and females (0.91; 95% CI, 0.85, 0.98) with CD. Adjustment for pubertal maturation did not alter the estimate; however, addition of lean mass to the model eliminated the bone deficit. Steroid exposure was associated with short stature but not bone deficits. CONCLUSION: This study shows the importance of considering differences in body size and composition when interpreting DXA data in children with chronic inflammatory conditions and shows an association between deficits in muscle mass and bone in pediatric CD. PMID- 15537439 TI - Osteoporosis prevalence and levels of treatment in primary care: the Australian BoneCare Study. AB - The level of recognition and treatment of osteoporosis is not well characterized in primary care. In data from a large sample of postmenopausal women attending 927 primary care physicians, 29% of women reported one or more fractures after menopause. The great majority (72%) were not on any osteoporosis-specific therapy. INTRODUCTION: Osteoporosis is often first recognized at the time of a low-trauma fracture. However, by this stage, the risk of subsequent fractures has already risen substantially. Moreover, in many countries, only a small proportion of women, who have already sustained fractures, receive a treatment shown to reduce this increased risk of further fractures. MATERIALS AND METHODS: This project was initiated to examine the prevalence of osteoporotic fractures, risk factors for osteoporosis, and use of antifracture therapy among postmenopausal Australian women. More than 88,000 women from 927 primary care physicians returned over 69,358 surveys. Of these, 57,088 reported the presence of a postmenopausal fracture or risk factors. RESULTS: Among these randomly selected postmenopausal women, 29% reported having had one or more low-trauma fractures after menopause (44% substantiated in current records). One-third of these women reported multiple fractures. The prevalence of all types of fractures, except rib and ankle, increased with age and low body weight. Those who reported fractures were also more likely to report early menopause, corticosteroid use, and a family history of osteoporosis. Moreover, those with vertebral fractures were more likely to record height loss, kyphosis, and back pain. Physical inactivity, low calcium intake, and smoking had no consistent relationship with any fracture outcome. Of the women who reported a fracture after menopause, only 28% were on any specific therapy for osteoporosis, and 7% were on calcium alone. Of those who had been told they had osteoporosis by a doctor, 40% were receiving specific osteoporosis therapy. CONCLUSIONS: In this large study of postmenopausal Australian women attending primary care physicians, 29% reported at least one low trauma fracture after menopause. Less than one-third of these women were on specific treatment for osteoporosis, and only 40% were ever told they had osteoporosis. Therefore, osteoporotic fractures are common in postmenopausal Australian women, and few, despite their substantially increased risk of further fractures, are on any specific anti-osteoporotic therapy. These data support the need for more effective education for the community and medical practitioners of the clinical significance of osteoporotic fractures and alternatives for treatment. PMID- 15537440 TI - Epidemiology of childhood fractures in Britain: a study using the general practice research database. AB - A population-based British cohort study, including approximately 6% of the population, was used to derive age- and sex-specific incidence rates of fractures during childhood. Fractures were more common among boys than girls, with peak incidences at 14 and 11 years of age, respectively. At childhood peak, incidence rates were only surpassed later in life at 85 years of age among women and never among men. INTRODUCTION: Fractures account for 25% of accidents and injuries in childhood; however, the descriptive epidemiology of childhood fractures remains uncertain. MATERIALS AND METHODS: Age- and sex-specific incidence rates for fractures at various skeletal sites were derived from the General Practice Research Database (a population-based British cohort containing computerized medical records of approximately 7,000,000 residents) between 1988 and 1998. RESULTS: A total of 52,624 boys and 31,505 girls sustained one or more fractures over the follow-up period, for a rate of 133.1/10,000 person-years. Fractures were more common in boys (161.6/10,000 person-years) than girls (102.9/10,000 person-years). The most common fracture in both sexes was that of the radius/ulna (30%). Fracture incidence was greater among boys than girls at all ages, with the peak incidence at 14 years of age among boys and 11 years of age among girls. Marked geographic variation was observed in standardized fracture incidence, with significantly (p < 0.01) higher rates observed in Northern Ireland, Wales, and Scotland compared with southeast England. CONCLUSIONS: Fractures are a common problem in childhood, with around one-third of boys and girls sustaining at least one fracture before 17 years of age. Rates are higher among boys than girls, and male incidence rates peak later than those among females. At their childhood peak, the incidence of fractures (boys, 3%; girls, 1.5%) is only surpassed at 85 years of age among women and never among men. The most common site affected in both genders is the radius/ulna. Studies to clarify the pathogenesis of these fractures, emphasizing bone fragility, are now required. PMID- 15537441 TI - When should the doctor order a spine X-ray? Identifying vertebral fractures for osteoporosis care: results from the European Prospective Osteoporosis Study (EPOS). AB - Vertebral fractures are common but usually remain unrecognized in primary care. Data from 2908 women and 2653 men in the EPOS study were used to derive algorithms to indicate the need for a spine X-ray to identify a fracture using easily elicited determinants. At a sensitivity of 50% for identifying cases, the specificity was increased from 50% to 78% in women and from 50% to 72% in men compared with a random allocation of X-rays. Use of X-rays can be optimized by selecting patients at high risk using a short screening procedure. INTRODUCTION: Previous osteoporotic fracture is an independent risk factor for further fractures and an indication for treatment. Vertebral fractures are the most common osteoporotic fractures before age 75, accounting for 48% of all fractures in men and 39% in women over 50. They usually remain unrecognized, so many patients requiring treatment are denied it, doubling their risk of a further fracture. Our objective was to develop an efficient algorithm indicating the need for an X-ray. MATERIALS AND METHODS: Data from 2908 women and 2653 men >or=50 years of age in the European Prospective Osteoporosis Study (EPOS) were analyzed. Lateral thoracic and lumbar spine radiographs were taken at baseline and at an average of 3.8 years later. Prevalent fractures were qualitatively diagnosed by an experienced radiologist. Fracture risk was modeled as a function of age, statural height loss since age 25, gender, and fracture history including limb fractures in the last 3 years using negative binomial regression. Receiver operating characteristic (ROC) curves were used to summarize a model's predictive ability, and a prediction algorithm was devised to identify those most likely to have a fracture. RESULTS: In a multivariate model for women, the risk of prevalent vertebral fracture significantly increased with age (RR, 1.67 [95% CI, 1.46, 1.93] per decade), statural height loss (1.06, [1.03, 1.10] per centimeter decrease), self-reported history of spine fracture (7.52 [5.52, 10.23]), and history of other major fracture (1.83 [1.46, 2.28]). Higher body weight reduced risk (0.86 [0.79, 0.95] per 10-kg increase). In men, the respective RR estimates were as follows: age (1.32 [1.18, 1.49]); height loss (1.06 [1.04, 1.09]); self reported spine fracture (5.05 [3.69, 6.90]); other major fracture (1.42 [1.12, 1.81]); and weight (0.86 [0.79, 0.94]). Using algorithms based on these easily elicited determinants, specificity was increased from 50% to 78% in women and from 50% to 72% in men at a sensitivity of 50% compared with a random allocation of X-rays. At a sensitivity of 75%, the specificity was 50% in women and 40% in men. Inclusion of hip BMD (femoral neck or trochanter), measured in 1360 women and 1046 men, significantly improved the area under the ROC curves by 4% in women (p < 0.002) but not in men (p > 0.350). Spine BMD, measured in 982 women and 847 men, produced a significant 5% AUC improvement in women (p = 0.007) but not in men (p = 0.554). CONCLUSION: A woman 65 years of age with one vertebral fracture has a one in four chance of another fracture over 5 years, which can be reduced to one in eight by treatment. Positive treatment decisions are often contingent on identifying a vertebral fracture. Selective use of lateral vertebral X-rays can be optimized using a 2-minute screening procedure administered by a nurse. PMID- 15537442 TI - Disc space narrowing is associated with an increased vertebral fracture risk in postmenopausal women: the OFELY Study. AB - We have analyzed the relationship between spine osteoarthritis and fractures in the OFELY cohort. Despite a higher BMD associated with spine OA, the risk of fragility fractures is not reduced. Disc space narrowing is associated with an increased risk of vertebral fracture. These data indicate that the risk of osteoporotic fracture should not be underestimated in women with spine OA. INTRODUCTION: Although osteoarthritis (OA) and osteoporosis both increase with age, their co-existence is uncommon. A higher BMD in OA is well documented, but a reduction of the fracture risk is still controversial. Our objective was to analyze the risk of fracture in postmenopausal women with spine OA. MATERIALS AND METHODS: In a cross-sectional study, spine OA was evaluated by lateral radiographs according to the method of Lane, and BMD was measured by DXA in 559 postmenopausal women from the OFELY cohort (mean age, 68 +/- 8 years; range, 58 94 years) 8 years after their inclusion into the study. Previous fragility fractures, all confirmed by radiographs, were prospectively registered during the annual follow-up for 8 years, and vertebral fractures were evaluated with spine radiographs. Severity of OA was assessed by scoring on osteophytes and disc narrowing on a four-point scale from 0 (normal) to 3 (severe) and graded as 0 (normal), 1 (mild osteophyte and/or narrowing), or 2 (moderate or severe osteophyte and/or narrowing). RESULTS: Osteophytes and disc narrowing were present in 75% and 64%, respectively, of women at the lumbar spine and in 88% and 51%, respectively, at the thoracic spine, increasing with age. BMD of the spine, hip, and whole body increased with the severity of osteophytosis, whereas severity of narrowing was associated with a higher BMD only at the spine. Ninety six fractures, including 48 vertebral fractures, occurred before OA assessment. No significant association was found between spine OA and all fragility fractures. In contrast, disc narrowing was associated with an increased risk of vertebral fracture with an odds ratio (95% CI) of 3.2 (1.1-9.3) after adjusting for age, body mass index, and BMD. The risk of vertebral fracture increased with the severity of disc narrowing. In comparison with the score 0, the odds ratio increased from 2.8 (0.9-8.7) to 4.6 (1.2-16.9) in women with mild to severe disc narrowing score. CONCLUSIONS: Despite a higher BMD, women with spine OA do not have a reduced risk of fracture. Disc narrowing is associated with a significant increased vertebral fracture risk. PMID- 15537443 TI - Bone fragility and collagen cross-links. AB - Infrared imaging analysis of iliac crest biopsy specimens from patients with osteoporotic and multiple spontaneous fractures shows significant differences in the spatial variation of the nonreducible:reducible collagen cross-links at bone forming trabecular surfaces compared with normal bone. INTRODUCTION: Although the role of BMC and bone mineral quality in determining fracture risk has been extensively studied, considerably less attention has been paid to the quality of collagen in fragile bone. MATERIALS AND METHODS: In this study, the technique of Fourier transform infrared imaging (FTIRI) was used to determine the ratio of nonreducible:reducible cross-links, in 2- to 4-microm-thick sections, from human iliac crest biopsy specimens (N = 27) at bone-forming trabecular surfaces. The biopsy specimens were obtained from patients that had been diagnosed as high- or low-turnover osteoporosis, as well as premenopausal women <40 years of age, with normal BMD and biochemistry, who suffered multiple spontaneous fractures. The obtained values were compared with previously published analyses of trabecular bone from normal non-osteoporotic subjects (N = 14, 6 males and 8 females; age range, 51-70 years). RESULTS AND CONCLUSIONS: Collagen cross-links distribution within the first 50 microm at forming trabecular surfaces in patients with fragile bone was markedly different compared with normal bone. PMID- 15537444 TI - The COMT val158met polymorphism is associated with peak BMD in men. AB - The associations between the functional val158met polymorphism of the estrogen degrading COMT enzyme and skeletal properties in young men were investigated. BMD was associated with COMT genotype. INTRODUCTION: Peak BMD is an important predictor of future risk of osteoporosis, and it is to a large extent determined by genetic factors. Estrogens are involved in the accretion of bone mass during puberty. Catechol-O-methyltransferase (COMT) is involved in the degradation of estrogens. There is a functional polymorphism in the COMT gene (val158met), resulting in a 60-75% difference in enzyme activity between the val (high activity [H]) and met (low activity [L]) variants. The aim of this cross sectional study was to investigate the associations between this polymorphism and peak BMD in young men. MATERIALS AND METHODS: A total of 458 healthy men (mean age, 19 +/- 0.6 years) were genotyped and classified as COMT(LL), COMT(HL), or COMT(HH). Areal BMD (aBMD) was measured by DXA. Cortical and trabecular volumetric BMD (vBMD) were measured by pQCT. The associations between COMT genotype and skeletal phenotypes were determined. RESULTS AND CONCLUSIONS: Regression models using physical activity, height, weight, age, and COMT genotype as covariates showed that COMT genotype was an independent predictor of aBMD in the total body and in all femur locations investigated, but not in the spine. The values for COMT(HL) and COMT(HH) were very similar, and therefore, they were pooled into one group. aBMD at Ward's triangle, trochanter, and total femur were 4.9%, 4.5%, and 3.7% lower, respectively, in the COMT(LL) than in the COMT(HL/HH) group (p < 0.01). pQCT analyses showed that COMT genotype was an independent predictor of trabecular vBMD of the tibia, radius, and fibula. Trabecular vBMD of the radius and fibula in COMT(LL) was 5.3% and 7.4% lower, respectively, than that of the combined COMT(HL/HH) group. COMT genotype was associated with cortical vBMD but not with cortical cross-sectional area in the tibia. These findings show that the COMT polymorphism is associated with BMD in young adult men. PMID- 15537445 TI - Strontium ranelate improves bone resistance by increasing bone mass and improving architecture in intact female rats. AB - Strontium ranelate given to intact rats at doses up to 900 mg/kg/day increases bone resistance, cortical and trabecular bone volume, micro-architecture, bone mass, and total ALP activity, thus indicating a bone-forming activity and an improvement of overall bone tissue quality. INTRODUCTION: Various anti osteoporotic agents are available for clinical use; however, there is still a need for drugs able to positively influence the coupling between bone formation and bone resorption to increase bone mass and bone strength. Strontium ranelate (PROTELOS), a new chemical entity containing stable strontium (Sr), was tested for its capacity to influence bone quality and quantity. MATERIALS AND METHODS: The long-term effects of strontium ranelate on bone were investigated in intact female rats treated with various doses of strontium ranelate (0, 225, 450, and 900 mg/kg/day) for 2 years. In a second series of experiments, the effects of 625 mg/kg/day were evaluated in intact male and female rats for the same period of time. Bone mineral mass and mechanical properties were evaluated at various skeletal sites (vertebra and femur), and bone tissue micro-architecture was evaluated by static histomorphometry at the tibio-fibular junction (cortical bone) and at the tibia metaphysis (trabecular bone). Plasma total alkaline phosphatase (ALP) activity and serum levels of insulin-like growth factor-I (IGF I) were also assessed. RESULTS: In female rats treated with strontium ranelate over 2 years, dose-dependent increases of bone strength and bone mass of the vertebral body (containing a large proportion of trabecular bone) and of the midshaft femur (containing mainly cortical bone) were detected without change in bone stiffness. Similar effects were observed in males at the level of the vertebra. This increase in mechanical properties was associated with improvements of the micro-architecture as assessed by increases of trabecular and cortical bone volumes and trabecular number and thickness. Finally, plasma total ALP activity and IGF-I were also increased in treated animals, compatible with a bone forming activity of strontium ranelate. CONCLUSION: A long-term treatment with strontium ranelate in intact rats is very safe for bone and improves bone resistance by increasing bone mass and improving architecture while maintaining bone stiffness. PMID- 15537446 TI - Combined bone morphogenetic protein-2 and -7 gene transfer enhances osteoblastic differentiation and spine fusion in a rodent model. AB - To enhance the osteogenic activity of BMP, combination BMP2 and BMP7 gene transfer was performed. This approach led to a significant increase in osteoblastic differentiation of mesenchymal precursors compared with single BMP gene transfer in vitro. When tested in 78 rats, combination gene transfer enhanced mechanically stable spine fusion and bone formation rate versus single BMP gene transfer. INTRODUCTION: Although clinical bone morphogenetic protein (BMP) therapy is effective, required doses are very high. Previous studies have suggested that the co-expression of two different BMP genes can result in the production of heterodimeric BMPs that may be more potent than homodimers. In this study, combined BMP2 and BMP7 gene transfer was performed to test whether this approach improves osteoblastic differentiation and bone formation compared with single BMP gene transfer. MATERIALS AND METHODS: A producer cell (A549) was co transfected with adenovirus vectors encoding BMP2 (AdBMP2) and BMP7 (AdBMP7) or, as controls, each vector alone, AdNull (with no transgene) or no virus. Supernatants were compared for their ability to stimulate osteoblastic differentiation of C2C12 myoblasts and MC3T3-E1 pre-osteoblasts. In a rat posterolateral spine fusion model, co-administration of AdBMP2 and AdBMP7 was compared with treatment with each vector alone, AdNull or no virus in 78 rats. The spines were assessed 8 weeks after surgery for radiographic and mechanical fusion, bone formation, and mineralization. RESULTS: BMP2 and BMP7 were co precipitated from supernatants of cells co-transfected with AdBMP2 and AdBMP7, indicating the presence of BMP2/7 heterodimer. Supernatants of co-transfected cells containing relatively low doses (7-140 ng/ml) of BMPs induced osteocalcin expression and alkaline phosphatase activity in both C2C12 and MC3T3-E1 cells, that were up to 6- and 40-fold higher, respectively, than levels induced by maximal doses (200-1000 ng/ml) of either BMP2 or BMP7 alone. In the spine fusion model, co-administration of AdBMP2 and AdBMP7 resulted in a significantly greater number of mechanically stable fusions and also 2-fold higher mineralization rate and bone volume in the fusion mass versus single BMP gene transfer (p < 0.02, all comparisons). CONCLUSION: Combined BMP2 and BMP7 gene transfer is significantly more effective in inducing osteoblastic differentiation and spine fusion than individual BMP gene transfer. PMID- 15537447 TI - Decreased BMD and limb deformities in mice carrying mutations in both Lrp5 and Lrp6. AB - Humans and mice lacking Lrp5 have low BMD. To evaluate whether Lrp5 and Lrp6 interact genetically to control bone or skeletal development, we created mice carrying mutations in both Lrp5 and the related gene Lrp6. We found that compound mutants had dose-dependent deficits in BMD and limb formation, suggesting functional redundancy between these two genes in bone and limb development. INTRODUCTION: Lrp5 and Lrp6 are closely related members of the low density lipoprotein receptor family and are co-receptors for Wnt ligands. While Lrp5 mutations are associated with low BMD in humans and mice, the role of Lrp6 in bone formation has not been analyzed. MATERIALS AND METHODS: To address whether Lrp5 and Lrp6 play complimentary roles in bone and skeletal development, we created mice with mutations in both genes. We inspected limbs of mice from the different genotypic classes of compound mutants to identify abnormalities. DXA and muCT were used to evaluate the effect of mutations in Lrp5 and Lrp6 on BMD and microarchitecture. RESULTS: Mice heterozygous for mutations in Lrp6 and either heterozygous or homozygous for a mutation in Lrp5 (Lrp6(+/-);Lrp5(+/-) or Lrp6(+/-);Lrp5(-/-)) display limb defects with incomplete penetrance and variable expression. DXA analysis showed that BMD decreased as mice progressively were more deficient in Lrp5 and Lrp6. Lrp6(+/-);Lrp5(-/-) mice were more severely affected than Lrp6(+/+);Lrp5(-/-) mice, whereas Lrp6(+/-);Lrp5(+/-) mice had statistically higher BMD than Lrp6(+/+);Lrp5(-/-) mice and lower BMD compared with wildtype mice and mice heterozygous for either mutation alone. CONCLUSIONS: Lrp6 and Lrp5 genetically interact in limb development in mice. Furthermore, heterozygosity for an inactivating mutation in Lrp6 further reduces BMD in both male and female mice lacking Lrp5. PMID- 15537448 TI - Increased VEGF expression in the epiphyseal cartilage after ischemic necrosis of the capital femoral epiphysis. AB - Ischemic injury to the immature femoral head produces epiphyseal cartilage damage and cessation of endochondral ossification. This study suggests that VEGF facilitates the repair of the necrotic epiphyseal cartilage, which is essential for restoration of endochondral ossification and re-establishment of the growth of the immature femoral head after ischemic necrosis. INTRODUCTION: Legg-Calve Perthes disease (LCPD) is a childhood form of osteonecrosis that produces growth arrest of the secondary center of ossification. The cessation of growth is caused by ischemic damage to the hypertrophic zone of the epiphyseal cartilage where endochondral ossification normally occurs. The role of vascular endothelial growth factor (VEGF) in restoring endochondral ossification in the epiphyseal cartilage after ischemic necrosis was investigated in a piglet model of LCPD because the resumption of normal growth is important for maintaining the spherical shape of the femoral head. MATERIALS AND METHODS: Piglet femoral heads were assessed 24 h to 8 weeks after the surgical induction of ischemia. Western blot analysis, ribonuclease protection assay (RPA), immunohistochemistry, and in situ hybridization were performed. RESULTS: Western blot analysis and RPA showed increased VEGF protein and mRNA expression, respectively, in the epiphyseal cartilage of the infarcted heads compared with the contralateral normal heads. In the normal femoral heads, VEGF-immunoreactivity (VEGF-IR) and transcripts were observed in the hypertrophic zone of the epiphyseal cartilage. In the infarcted heads, VEGF-IR and transcripts were no longer observed in the hypertrophic zone because of diffuse cell death in that zone from ischemia. However, VEGF-IR and transcripts were observed in the proliferative zone above the necrotic hypertrophic zone. At 8 weeks, vascular granulation tissue invasion of the necrotic hypertrophic zone was observed with active resorption of the necrotic cartilage. In some areas where the necrotic cartilage was completely resorbed, restoration of endochondral ossification was observed. In these areas, VEGF transcripts were observed in the newly formed hypertrophic zone. CONCLUSIONS: VEGF expression was increased, and its spatial expression was altered in the epiphyseal cartilage after ischemic necrosis of the immature femoral head. VEGF upregulation in the proliferative zone after ischemic damage may play a role in stimulating vascular invasion and granulation tissue formation in the necrotic hypertrophic zone of the epiphyseal cartilage. This may be an important step toward facilitating the resorption of the necrotic cartilage and restoration of endochondral ossification leading to further growth and development of the femoral head. PMID- 15537449 TI - Decreased bone formation and osteopenia in mice lacking alpha-calcitonin gene related peptide. AB - We recently described an unexpected high bone mass phenotype in mice lacking the Calca gene that encodes CT and alphaCGRP. Here we show that mice specifically lacking alphaCGRP expression display an osteopenia caused by a decreased bone formation. These results show that alphaCGRP is a physiological activator of bone formation and that the high bone mass phenotype of the Calca-deficient mice is caused by the absence of CT. INTRODUCTION: Calcitonin (CT) and alpha-calcitonin gene-related peptide (alphaCGRP) are two polypeptides without completely defined physiologic functions that are both derived from the Calca gene by alternative splicing. We have recently described an unexpected high bone mass phenotype in mice carrying a targeted deletion of the Calca gene. To uncover whether this phenotype is caused by the absence of CT or by the absence of alphaCGRP, we analyzed a mouse model, where the production of alphaCGRP is selectively abolished. MATERIALS AND METHODS: Bones from Calca(-/-) mice, alphaCGRP(-/-) mice, and their corresponding wildtype controls were analyzed using radiography, muCT imaging, and undecalcified histology. Cellular activities were assessed using dynamic histomorphometry and by measuring the urinary collagen degradation products. CT expression was determined using radioimmunoassay and RT-PCR. Immunohistochemistry was performed using an anti-CGRP antibody on decalcified bone sections. RESULTS: Unlike the Calca-deficient mice, the alphaCGRP-deficient mice do not display a high bone mass phenotype. In contrast, they develop an osteopenia that is caused by a reduced bone formation rate. Serum levels and thyroid expression of CT are not elevated in alphaCGRP-deficient mice. While CGRP expression is detectable in neuronal cell close to trabecular bone structures, the components of the CGRP receptor are expressed in differentiated osteoblast cultures. CONCLUSION: The discrepancy between the bone phenotypes of Calca(-/-) mice and alphaCGRP(-/-) mice show that the high bone mass phenotype of the Calca( /-) mice is caused by the absence of CT. The osteopenia observed in the alphaCGRP(-/-) mice that have normal levels of CT further show that alphaCGRP is a physiologic activator of bone formation. PMID- 15537450 TI - Regulation of human osteoclast differentiation by thioredoxin binding protein-2 and redox-sensitive signaling. AB - Differential expression of TBP-2 and Trx-1 occurs during osteoclastogenesis. Adenoviral overexpression of TBP-2 in osteoclast precursors inhibits Trx-1 expression, osteoclast formation, and AP-1 binding activity. TBP-2 and Trx-1 are key regulators of osteoclastogenesis. INTRODUCTION: Thioredoxin binding protein-2 (TBP-2) negatively regulates thioredoxin-1 (Trx-1), a key endogenous modulator of cellular redox and signaling. In gene array analysis, we found that TBP-2 expression was reduced during human osteoclast differentiation compared with macrophage differentiation. Our aim was to determine the roles of TBP-2 and Trx-1 in human osteoclastogenesis and RANKL signaling. MATERIALS AND METHODS: Osteoclasts or macrophages were generated from colony-forming unit-granulocyte macrophage (CFU-GM) precursors treated with sRANKL and macrophage-colony stimulating factor (M-CSF), or M-CSF alone, respectively. Expression of TBP-2 and Trx-1 was quantified by real-time PCR and Western analysis. Adenoviral gene transfer was used to overexpress TBP-2 in precursors. NF-kappaB and activator protein 1 (AP-1) signaling was assessed with EMSA. RESULTS: In the presence of sRANKL, expression of TBP-2 was decreased, whereas Trx-1 expression was increased. The antioxidant N-acetylcysteine reversed this pattern and markedly inhibited osteoclastogenesis. Adenoviral overexpression of human TBP-2 in precursors inhibited osteoclastogenesis and Trx-1 expression, inhibited sRANKL induced DNA binding of AP-1, but enhanced sRANKL-induced DNA binding of NF kappaB. CONCLUSIONS: These data support significant roles for TBP-2 and the Trx system in osteoclast differentiation that are mediated by redox regulation of AP 1 transcription. A likely mechanism of stress signal induction of bone resorption is provided. Modulators of the Trx system such as antioxidants have potential as antiresorptive therapies. PMID- 15537451 TI - CCR1 chemokines promote the chemotactic recruitment, RANKL development, and motility of osteoclasts and are induced by inflammatory cytokines in osteoblasts. AB - Chemoattractants that recruit OC precursors to locally inflamed sites of resorption are not well known. A chemokine receptor, CCR1, was expressed in OC precursors and elevated in mature OCs, and its ligands promoted OC precursor recruitment, RANKL development, and OC motility. Cytokines induced OB release of such chemokines, which may therefore significantly contribute to inflammatory bone loss. INTRODUCTION: Chemokines, primarily of two major (CXC, CC) families, are essential signals for the trafficking and localization of circulating hematopoietic cells into tissues. However, little is known about their potential roles in osteoclast (OC) recruitment, development, or function. Previously, we analyzed CXC receptors in murine OC precursors and found high expression of CXCR4 that mediated their stromal-derived factor-1(SDF-1)-induced chemotaxis and collagen invasion. Here, we investigated if CC receptors and ligands, which are elevated in inflammatory and other osteolytic diseases, also play important roles in the recruitment, formation, or activity of murine bone-resorptive OCs. MATERIALS AND METHODS: CC chemokine receptor (CCR) mRNA expression was analyzed during OC formation induced by RANKL in murine RAW 264.7 cells and primary marrow cells. Corresponding CC chemokines were tested for their ability to elicit precursor chemotaxis or OC development, or to influence motility, bone resorption, adhesion, or survival in RANKL-differentiated OCs. Constitutive and inflammatory cytokine-induced release of the chemokines macrophage inflammatory protein-1alpha (MIP-1alpha) and regulated on activation, normal T-cell expressed and secreted (RANTES) was measured by ELISA for OCs, osteoblasts (OBs), and their precursor cells. RESULTS: CCR1 was expressed in murine marrow cells, the most prominent CCR in RAW cells, and upregulated by RANKL in marrow or RAW cells. Chemokines that bind CCR1 (MIP-1alpha, RANTES, and monocyte chemoattractant protein-3 [MCP-3]) were produced to varying degrees by murine OCs, OBs, and their precursors, and markedly increased by interleukin (IL)-1alpha and TNFalpha in differentiating OBs. RANTES, and especially MIP-1alpha, increased mature OC motility, but did not alter OC resorption activity, adhesion, or survival. All three chemokines stimulated chemotaxis of marrow or RAW cell precursors, leading to the greater formation of OCs (in number and size) after RANKL development of such chemoattracted marrow cells. All three chemokines also directly and dramatically enhanced OC formation in marrow cultures, through a pathway dependent on the presence of RANKL but without altering RANK expression. CONCLUSIONS: Pathological increases in secretion of these chemokines from activated OBs or other cells may potently stimulate the chemotactic recruitment and RANKL formation of bone-resorptive OCs, thereby exacerbating local osteolysis in multiple skeletal diseases. PMID- 15537452 TI - Amino-terminal parathyroid hormone fragment analogs containing alpha,alpha-di alkyl amino acids at positions 1 and 3. AB - To define and minimize the N-terminal PTH pharmacophore, the effects of introducing different conformationally constraining di-alkyl amino acids at positions 1 and 3 of PTH(1-14) analogs were studied. Improvements in PTH receptor binding affinity and signaling potency were found, although some substitutions resulted in partial agonism. INTRODUCTION: The N-terminal portion of parathyroid hormone (PTH) plays a critical role in PTH-1 receptor (P1R) activation. To investigate the mechanisms underlying this action and to minimize the N-terminal PTH pharmacophore, we employed the PTH(1-14) fragment as a scaffold for structure activity relationship studies, and thus previously found that substitutions of the conformationally constraining, di-alkyl amino acid, alpha-amino-isobutyric acid (Aib), at positions 1 and 3 increase the P1R-binding affinity and signaling potency of the analog approximately 100-fold. Here we extend these findings by investigating the effects of other constrained di-alkyl amino acids at positions 1 and/or 3 on PTH analog activity. MATERIALS AND METHODS: The di-alkyl amino acids, 1-aminocycloalkane-carboxylic acid (Ac(x)c, x = 3, 5, or 6) or diethylglycine (Deg), representing alkyl configurations of varying volumes and shape (cyclic and linear), were introduced into the parent peptide, [M]PTH(1-14) (M = Ala(1,3,12),Gln(10),Har(11),Trp(14)), and the analogs were tested for activity in P1R-expressing cells. RESULTS: Relative to the binding affinity and cAMP-stimulating potency of the parent peptide (IC(50) = 27 mM; EC(50) = 220 nM), PTH(1-14) analogs substituted at position 1 exhibited 2- (Ac(3)c) to 60-fold (Ac(5)c) increases in affinity and potency, as measured in LLC-PK1 cells stably expressing the cloned P1R. Combining the substitutions of Ac(5)c(1) and Aib(3) yielded the highest affinity and most potent PTH(1-14) and shorter-length analogs to date: [Ac(5)c(1), Aib(3),M]PTH(1-X) (X = 14, 11, and 10; IC(50)s = 80 nM, 260 nM, and 850 microM; EC(50)s = 1.7 nM, 3.1 nM, and 1.9 microM, respectively). The effects of Ac(6)c(1) were similar to those of Ac(5)c(1). A dissociation of binding affinity and signaling activity occurred with Deg, as [Deg1,3,M]PTH(1-14) was a partial agonist. CONCLUSION: Constraining the N-terminal PTH backbone conformation with di-alkyl amino acids at positions 1 and 3 may be a general strategy for optimizing and minimizing the PTH pharmacophore; however, inhibitory side-chain effects may be encountered. The new analogs presented should be useful as minimum-length functional probes of the PTH-PTH receptor interaction mechanism. PMID- 15537453 TI - Antiretroviral therapy in sub-Saharan Africa: adherence lessons from tuberculosis and leprosy. AB - Declining drug costs and increases in international donor interest are leading to greater availability of antiretroviral treatment programmes for persons living with the human immunodeficiency virus in parts of sub-Saharan Africa. Ensuring adequate adherence to antiretroviral drug therapy is one of the principal challenges facing successful implementation in Africa, where 70% of the world's infected persons live. Tuberculosis and leprosy are two diseases of global importance whose control programmes can provide important lessons for developing antiretroviral drug adherence strategies. This paper examines various approaches used in tuberculosis and leprosy control which could help enhance adherence to antiretroviral therapy in resource-limited settings. PMID- 15537454 TI - Pigmented penile macules. AB - There are two opposing schools of thought regarding the management of pigmented penile macules. Allan and Spitz, as well as Pack and Davis, are of the opinion that almost all pigmented naevi present on the palms, soles and genitalia are junctional naevi and have a higher incidence of malignant change and, therefore, should be removed. On the other hand, because most cases of penile and urethral melanomas arise de novo with only a few arising from previous long-standing naevi and as precursor lesions are not identified in most cases, Scott et al. and Stegmaire et al. consider prophylactic excision unnecessary. A more practical approach has been taken up by Barnhill et al. and their view is that management should be individualized with multiple biopsies to establish the diagnosis. The purpose of this paper is to review the literature regarding the natural history, risk of melanoma development on the penis and the management of cases presenting with pigmented penile macules. PMID- 15537455 TI - Seeking and engaging in internet sex: a survey of patients attending genitourinary medicine clinics in Plymouth and in London. AB - We conducted a survey of patients attending two GUM clinics to evaluate the use of the internet for seeking sexual partners and for seeking on-line sexual health information. Prevalence of internet sex-seeking (ISS) was 11% (57/500) in Plymouth and 21% (50/237) in London. ISS was more common among men who have sex with men (MSM) (43/92 = 47%) than men who have sex with women (MSW) (38/280 = 14%, P < 0.0001) and more common amongst MSW than women who have sex with men (WSM) (25/350 = 7%, P = 0.007). In both samples, accessing sexual health information online was associated with ISS (Plymouth, P = 0.05 and London, P = 0.001), as was reporting more than 10 sexual partners per year (Plymouth and London, P < 0.001). It remains to be seen whether the potential sexual health promotion benefits of the internet could balance potential risks of STI acquisition. PMID- 15537456 TI - Managing genital chlamydial infection: the effects of ethnic origin and method of follow-up. AB - We assessed if ethnicity and method of follow-up were associated with differences in the number of patients and the number of partners successfully treated for genital chlamydial infection. This was done by retrospectively reviewing the first 400 cases of genital chlamydia diagnosed between February and June 2001 who had a traditional clinic follow-up and the first 400 cases from the same period in 2002 when the telephone follow-up was used. The telephone follow-up appointment system, when compared to a traditional clinic follow-up appointment system, eliminated differences between Black and White ethnic groups in the numbers of patients and partners satisfactorily treated. PMID- 15537457 TI - Post-HIV test counselling of clients of a mobile STD/HIV clinic. AB - The objective of the study was to determine the post-test counselling (PTC) rates for HIV-infected and uninfected individuals receiving HIV counselling and testing on a mobile STD/HIV screening clinic and to determine whether individuals at highest risk for transmitting their infection were less likely to receive PTC than those at lower risk for transmitting. Clients presenting for HIV counselling and testing were asked about their demographic characteristics, clinical history, personal risk behaviours, and partner risk factors and told to return after 14 days for results. Disease intervention specialists (DIS) attempted to locate and counsel positive clients. The PTC rate among infected and uninfected clients was 66% and 46%, respectively. There were significant differences in demographics and risk factors for those who were post-test counselled versus those who were not. Among HIV-uninfected clients, there was a positive association between PTC and drug treatment in the past three months and having engaged in sex work within the last three months. Being female was negatively associated with PTC. Among HIV infected clients, there was a positive association between PTC and current enrollment in drug treatment. These data suggest that mobile STD/HIV screening clinics may be limited in their effectiveness by low rates of PTC. PMID- 15537458 TI - Factors associated with sexual problems in HIV-positive gay men. AB - Our objective was to determine factors associated with sexual problems in a sample of HIV-seropositive gay male clinic patients. Using a cross-sectional survey design a volunteer sample of 78 outpatient HIV-seropositive gay male service users completed a self-report questionnaire. This examined sexual problems, their perceived causes and associated factors including demographics, health status, sexual behaviour, self-justifications for sexual risk-taking and mood state (Hospital Anxiety and Depression Scale). Fifty (69%) of 78 HIV positive gay men reported one or more sexual problems. Erectile dysfunction (ED) was reported by 38% rising to 51% in the context of trying to use condoms. Loss of interest in sex was reported by 41% and 24% experienced delayed ejaculation. The presence of sexual problems affected condom use in that 33 (90%) of the 37 gay men who had ED associated with condom use were inconsistent condom users in insertive sex compared to 28% of those not having this type of ED (P < 0.001). The presence of ED did not reduce the frequency of anal intercourse but those with ED associated with condoms were significantly more likely to have had receptive anal sex in the past three months (62%) compared to men without ED with condoms (38%) (P = 0.05). Risk cognitions such as wanting to lose oneself in sex, leaving responsibility for condom use to the active partner and perceptions that condoms interfere with pleasure were significantly more likely to be endorsed by those who report ED with condoms. Other factors associated with sexual problems included low T-cell counts (i.e. < 200). Psychological explanations were the most frequently cited causes of sexual problems, whether alone or in interaction with HIV disease itself, and combination therapy. A high incidence of sexual problems was found amongst this sample of HIV-positive gay men. Untreated sexual dysfunctions may contribute to sexual risk-taking and therefore HIV clinics need to address both issues. Further research is required to better understand the role of psychological factors, HIV disease itself and combination therapy in the incidence and treatment of sexual problems. PMID- 15537459 TI - Which treatment for genital tract Chlamydia trachomatis infection? AB - A national opportunistic chlamydia screening programme, mainly targeting young sexually active women, is gradually being introduced across the UK and in future will predominantly occur in primary care sites. The relative efficacy of recommended antibiotic treatments for chlamydia has been poorly studied and especially that of single dose azithromycin. In Portsmouth, 1536 patients treated for chlamydia, with four different antibiotic regimens, during the Department of Health pilot study, were asked to return for test of cure. No difference in treatment outcome was found using doxycycline, oxytetracycline, erythromycin or azithromycin. Directly observed therapy with azithromycin may be especially helpful in treating young chlamydia-positive patients. PMID- 15537460 TI - Detection of high-risk HPV DNA in semen and its association with the quality of semen. AB - The effects of seminal high-risk human papillomavirus (HPV) DNA were assessed on the quality of semen. Semen samples of 65 men participating in the ongoing Finnish HPV Family Study were collected. Semen analyses were done by the guidelines of the Nordic Association for Andrology. HPV DNA was detected by nested polymerase chain reaction and confirmed by Southern blot hybridization for high-risk types. Altogether, 10/65 men (15.4%) had high-risk HPV DNA positive semen sample. Seminal high-risk HPV DNA did not affect semen volume, sperm concentration, motility and vitality of spermatozoa. However, semen pH was borderline lower in HPV DNA positive than negative samples (7.4 vs 7.5). Neither oligo- nor asthenozoospermia was associated with seminal HPV DNA. In conclusion, seminal high-risk HPV DNA was detected in 15% of men. It did not affect the semen analysis, except semen pH by borderline significance. Sperm donors have not been tested for HPV infections, sperm washing does not seem to eliminate the risk of HPV transmission and the consequences of HPV in the semen are at present unknown. PMID- 15537461 TI - Re-infection of Chlamydia trachomatis in patients presenting to the genitourinary medicine clinic in Portsmouth: the chlamydia screening pilot study - three years on. AB - Chlamydia trachomatis is the most common bacterial sexually transmitted infection (STI) in the UK. The Department of Health set up an opportunistic screening programme for genital chlamydia infection, focusing on sexually active 16-24-year old women and some men. This study identified those patients re-attending the genitourinary medicine (GUM) clinic and followed them up until September 2002. We examined the reasons for attendance and re-infection with chlamydia. Two hundred and eighty-five patients re-attended the clinic. Two-thirds of these had changed their sexual partners in the follow-up period. Fifty-six patients were diagnosed with genital chlamydia infection in subsequent clinic visits. The majority of them had changed their sexual partners, suggesting new acquisition of infection. This suggests that in this age group re-screening should be offered at a year interval. Patients diagnosed with genital chlamydia infection should be referred to the GUM clinic for further STI screening and partner notification. PMID- 15537462 TI - Health literacy and patient knowledge in a Southern US HIV clinic. AB - Pharmaceutical management of HIV infection is complex, and proper adherence to antiretroviral regimens is contingent on active patient involvement in treatment. We examined the relationship between patient understanding of HIV and its treatment and health literacy. Structured interviews were conducted with 157 HIV infected individuals receiving care at a community-based clinic in Shreveport, Louisiana, USA. In all, 48% of patients were reading below a 9th grade level. One third of patients could not name their HIV medications and this was significantly related to low literacy (P < 0.01). Two-thirds of those reading below the 9th grade level did not know how to take their medications correctly (P < 0.05), and 75% did not know the meaning of a CD4 count or viral load (P < 0.001). Patients with low literacy skills were more likely to state that their physician was their sole source of HIV information (P < 0.005). Physicians may require training to appropriately convey health information to patients of low literacy. PMID- 15537463 TI - Changing pattern of HIV infection in a tertiary care hospital in New Delhi, India. AB - HIV seroprevalence was carried out in 42,738 individuals attending the STD Centre of a New Delhi hospital from September 1990 to December 2001. The different epidemiological parameters of the patients in Group 1 (asymptomatic HIV seropositive individuals) and Group 2 (AIDS cases), were compared in four spans A, B, C, D. Significant rise in seroprevalence from 0.4% to 4.4% was observed with time. The patients in Group 1 were mainly 15-19 years followed by 30-44 years age group. Overall, the Male:Female ratio in Groups 1 and 2 were 3:1 and 6:1, respectively. The transmission was predominantly heterosexual in both the groups. The patients with sexually transmitted infections (STIs) emerged as the most prominent category in Group 1, showing a steady rising trend till 1999 and stabilizing thereafter. Ulcerative STIs, mainly syphilis, showed maximum association. In most of the cases, the infection was acquired by promiscuous males from female commercial sex workers or casual acquaintances and further transmitted to their spouses. Tuberculosis was the most common opportunistic infection. However, patients presented with fever, loss of weight and diarrhoea in increasing number during time D. The present study reiterates the importance of early management of STI patients and counselling of high risk groups, early partner notification and routine antenatal HIV check-up in the prevention and control of HIV/AIDS infection in third world countries like India. PMID- 15537464 TI - Opportunistic infections in hospitalized HIV-infected adults in Ho Chi Minh City, Vietnam: a cross-sectional study. AB - The HIV epidemic is emerging rapidly in Vietnam. We studied the prevalence of opportunistic infections by performing clinical and microbiological investigations in 100 hospitalized HIV-infected adults in Ho Cho Minh City, Vietnam. The median CD4 count was 20 cells/mm(3) and in-hospital mortality was 28%. The most frequent diagnoses were oral candidiasis (54), tuberculosis (37), wasting syndrome (34), lower respiratory tract infection (13), cryptococcosis (9), and penicilliosis (7). Bacterial (other than tuberculosis) and parasitic infections were uncommon. Regional differences should be considered when deciding which diagnostic procedures and prophylactic measures to implement. In Vietnam, routine mycobacterial blood cultures do not provide greater yield than chest radiography and sputum and lymph node aspirate smears. Prophylactic trimethoprim/sulphamethoxazole against Pneumocystis jiroveci pneumonia may confer little benefit, and high rates of isoniazid resistance may affect the efficacy and feasibility of tuberculosis chemoprophylaxis. However, the usefulness of itraconazole prophylaxis for cryptococcosis and penicilliosis merits further consideration. PMID- 15537465 TI - Completeness and timeliness of treatment for chlamydia within a sexual health service. AB - Prompt treatment of genital Chlamydia trachomatis infection will reduce the duration of infectiousness and further transmission of infection. In 2002, an audit was conducted to assess the completeness and timeliness of treatment for uncomplicated chlamydial infection at the Sydney Sexual Health Centre, Australia. Over a nine-month period, chlamydia was diagnosed in 250 patients, 97% of whom received appropriate treatment. The median duration between initial attendance and treatment was two days. Prompt delivery of treatment was facilitated by: initial treatment of patients likely to be chlamydia-infected; rapid availability of test results; urgent communication of positive test results; and effective procedures for recalling untreated patients. PMID- 15537466 TI - Management of HIV-related lymphoma in HIV treatment centres in North Thames Region. AB - We describe an audit of lymphoma care in HIV-positive patients. In the smaller units (< 500 patients) areas of potentially sub-optimal care were identified. PMID- 15537467 TI - Getting to the bottom of anal itch - a cautionary tale. AB - Peri-anal rash is common and may appear macroscopically benign. In HIV-positive men, however, more serious pathology may be revealed by a biopsy. The high prevalence of human papillomavirus infection in these men predisposes them to pre cancerous lesions of the peri-anal and anal skin. We report an unusual case of dual pathology in the anal region, which highlights the need to thoroughly investigate peri-anal symptoms in HIV-positive men. PMID- 15537468 TI - Oral Kaposi's sarcoma in a non-HIV homosexual White male. AB - This paper reports the exceedingly rare occurrence of oral Kaposi's sarcoma in a non-HIV Caucasian White male. The lesion presented as a 8 mm vascular nodule on the buccal mucosa on the left. He was homosexual. The lesion was removed and histological examination confirmed Kaposi's sarcoma. The patient has been tested on 10 occasions for HIV from December 1998 to March 2004 and has been negative. The patient remains alive and well with no evidence of recurrence. PMID- 15537469 TI - Incomplete reporting of race and ethnicity in gonorrhoea cases and potential bias in disease reporting by private and public sector providers. PMID- 15537470 TI - GU medicine consultants' and clients' opinions on general practitioner involvement with sexual health care. PMID- 15537471 TI - The impact of insulin glargine on clinical and humanistic outcomes in patients uncontrolled on other insulin and oral agents: an office-based naturalistic study. AB - OBJECTIVE: Controlling blood glucose levels in patients with diabetes often requires aggressive treatment, which may in turn cause hypoglycemia and/or decreased health-related quality of life (HRQOL). Insulin glargine, a long-acting insulin, has shown benefits of decreased nocturnal hypoglycemia without significant weight gain, while providing good glycemic control in clinical trials. These benefits have often been reported in studies of less than 1 year duration. The objective of this study was to evaluate the effectiveness of insulin glargine over a 12-month period in a clinical practice setting, and measure its effects on HRQOL in a subset of patients. DESIGN AND METHODS: Patients with diabetes in a large private endocrinology practice were initiated on insulin glargine. Patients were divided into 2 cohorts: the first group included patients with type 1 diabetes (T1D, n = 135); the second group included patients with type 2 diabetes previously on insulin and/or oral agents (T2D, n = 180). The HRQOL subset analysis included 50 patients from the above study. Patients completed a 40-item questionnaire adapted from the Diabetes Symptom Checklist-Revised (DSC-R) and the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36) at weeks 0, 2, 6, 12, and 16. RESULTS: Patients in both cohorts experienced statistically significant decreases in mean ( +/- SD) A1C: group T1D, -0.28 +/- 1.47 (P = 0.0307); group T2D, -0.60 +/- 1.51 (P < 0.0001), with no significant changes in body mass index. In the year following insulin glargine therapy, there were significantly fewer hypoglycemic events per patient than in the year prior to insulin glargine therapy (group T1D: -0.33, P = 0.002; group T2D: -0.20, P = 0.004). HRQOL subset analysis also revealed a significant decrease in A1C (P < 0.0001) after 16 weeks of therapy with insulin glargine. In this subset of patients, there was a significant improvement in overall well being (P = 0.0019), emotional well being (P = 0.003), total symptom scores (P < 0.0001), and total symptom distress (P < 0.0001). The limitations of the study are those inherently associated with naturalistic observational studies such as recall bias and compliance. CONCLUSIONS: Insulin glargine use over a 12-month period in a clinical practice setting was shown to significantly improve A1C without adversely impacting weight or the occurrence of hypoglycemia. Significant improvements were also observed in HRQOL. PMID- 15537472 TI - Combination therapy in asthma--fixed or variable dosing in different patients? AB - The introduction of combination products, for the coadministration of an inhaled corticosteroid (ICS) with a long-acting beta2-agonist in a single inhaler, has greatly simplified asthma therapy. The two combination inhalers currently available, Symbicort (budesonide/formoterol in a single inhaler) and Seretide (salmeterol/fluticasone), comply with Step 3 of international guidelines that recommend the addition of a long-acting beta2-agonist to ICS in patients who are inadequately controlled on ICS alone. Importantly, combination inhalers ensure that patients cannot neglect their ICS maintenance therapy in favour of the long acting beta2-agonist--which may improve adherence and overall asthma control. In vitro experiments suggest that ICS and long-acting beta2-agonists may interact beneficially when they are administered via one inhaler. The efficacy and tolerability of budesonide/formoterol and salmeterol/fluticasone have been demonstrated. There are currently two approaches for treating asthma using combination therapy--fixed and adjustable dosing. Fixed dosing with budesonide/formoterol or salmeterol/fluticasone provides effective asthma control in line with guideline goals. However, given the inherent variability of asthma, there is increasing evidence that adjusting the dose of ICS according to fluctuations in symptoms is beneficial. Findings from a series of studies comparing fixed and adjustable symptom-guided dosing regimens demonstrate that adjustable dosing may improve asthma control at an overall lower steroid dose. Ultimately, if adjustable dosing proves to be an effective treatment option, it may be possible to use budesonide/formoterol for both maintenance therapy and symptom relief, thereby overcoming the need for a separate reliever inhaler. This is because formoterol has a more rapid onset and greater dose-related effects than salmeterol in salmeterol/fluticasone. Given that all patients are different, with different disease severities and treatment preferences, both fixed and adjustable dosing strategies are likely to be important in the long-term management of asthma. It is possible that different treatment options will be used for different patients, depending on their disease severity, personality and ability to adhere to therapy. PMID- 15537473 TI - Cost-effectiveness of detemir-based basal/bolus therapy versus NPH-based basal/bolus therapy for type 1 diabetes in a UK setting: an economic analysis based on meta-analysis results of four clinical trials. AB - BACKGROUND: A meta-analysis of results from four clinical trials in type 1 diabetes patients showed that insulin detemir (IDet)-based basal/bolus treatment of type 1 diabetes led to improved HbA1c (0.15%-points lower), reduced risk of major hypoglycaemic events (by 2%) and reduction in body mass index (BMI) (0.26 kg/m2) compared to protamine Hagedorn human (NPH) insulin-based basal/bolus therapy in type 1 patients. METHODS: A published, validated, peer-reviewed Markov simulation model (the CORE Diabetes Model) projected short-term results obtained from the fixed-effects (weighted average) meta-analysis to long-term incidence of complications, improvements in quality-adjusted life years (QALY), long-term costs and the cost-effectiveness for IDet combinations versus NPH combinations in type 1 diabetes patients. Probabilities of complications and HbA1c-dependent adjustments were derived from the DCCT and other studies. Costs of treating complications in the UK were retrieved from published sources. Total direct costs (complications + treatment costs) for each arm were projected over patient lifetimes from a UK National Heath Service perspective. Both costs and clinical outcomes were discounted at 3.5% annually. RESULTS: Improved glycaemic control, decreased hypoglycaemic events and BMI with IDet-based basal/bolus therapy led to fewer diabetes-related complications, an increase in quality-adjusted life expectancy of 0.09 years, increased total lifetime costs/patient of 1707 pounds sterling and an incremental cost-effectiveness ratio of 19,285 pounds sterling per QALY gained. Results were stable under a wide range of reasonable assumptions. CONCLUSIONS: Short-term improvements seen with IDet combinations versus NPH combinations led to decreased complications, improvements in QALYs and reductions in complication costs, which partially offset the additional costs of detemir, leading to a cost-effectiveness ratio which fell within a range considered to represent excellent value for money (< 35,000 pounds sterling/QALY gained). PMID- 15537474 TI - An open-label, comparative study of rivastigmine, donepezil and galantamine in a real-world setting. AB - OBJECTIVE: We analysed the effects of donepezil, rivastigmine and galantamine, prescribed for the treatment of Alzheimer disease in a real-world setting in Italy. METHODS: Outcome measures included the MiniMental State Examination (MMSE), the Alzheimer Disease Assessment Scale cognitive subscale (ADAS-cog), Instrumental Activity of Daily Living (IADL) and ADL scales. RESULTS: Seventy patients were treated with donepezil, 121 with rivastigmine and 51 with galantamine. At 6 months, rivastigmine-treated patients improved by 1.29 points from baseline on the ADAS-cog, while donepezil- and galantamine-treated patients showed 'no change' (changes of < 0.2 points). On the IADL, patients treated with rivastigmine, donepezil and galantamine showed decreases of 0.42, 0.58 and 0.75 points, respectively. On the ADL, donepezil- and galantamine-treated patients showed decreases of 0.44 and 0.86 points, respectively, while there was 'no change' with rivastigmine. On the MMSE, donepezil- and rivastigmine-treated patients showed 'no change' and galantamine-treated patients showed a mean decrease of 1.19 points. A subgroup analysis of 'pseudo-randomised' patients (rivastigmine, n = 63; donepezil, n = 55; galantamine, n = 51) supported the main findings. Side effects were similar (in type and frequency) in the three treatment groups. CONCLUSIONS: This is the first study to compare the effects of the three most commonly-used cholinesterase inhibitors on the MMSE, ADAS-cog, IADL and ADL. Limitations included its small population size, its open-label design, and the fact that patients were randomised only after the introduction of galantamine. There were no statistically significant differences between the three drugs at 3 months. While numerical trends were observed suggesting the effect of rivastigmine > donepezil > galantamine, larger, longer-term prospective studies are needed to confirm whether there are important differences in the long term efficacy of the three drugs. PMID- 15537475 TI - A comparison of the fourth-generation fluoroquinolones gatifloxacin 0.3% and moxifloxacin 0.5% in terms of ocular tolerability. AB - PURPOSE: To compare the ocular tolerability of the commercially available ophthalmic solutions of the fourth-generation fluoroquinolones, gatifloxacin 0.3% (Zymar, Allergan, Inc., Irvine, CA) with benzalkonium chloride (BAK) and moxifloxacin 0.5% (Vigamox) without BAK. METHODS: A baseline evaluation was conducted on 30 healthy volunteers for conjunctival hyperemia, conjunctival vascularity, pupil size, and anterior chamber (AC) cell and flare. Pupils were measured under scotopic conditions with a Colvard pupillometer. Conjunctival hyperemia and vascularity, and AC reaction were measured on a Likert-like scale of 0-3. Subjects then received drops in both eyes from masked bottles of gatifloxacin ophthalmic solution 0.3% with BAK (in one eye determined randomly) and moxifloxacin ophthalmic solution 0.5% without BAK (in the contralateral eye) in a double-masked fashion. Subjects graded pain and ocular irritation in each eye on a scale of 1-10 after 5 min with their eyes closed. The examination was then repeated. RESULTS: The average age of this study population was 34.4 years. The groups of eyes receiving moxifloxacin 0.5% demonstrated an increase in mean conjunctival hyperemia (0.21 [range: 0-1] at baseline to 1.52 [range: 0-3] at 5 min.) that was significantly greater (p = 0.0005) compared with that of the group receiving gatifloxacin 0.3% (0.22 [range: 0-1] at baseline to 0.45 [range: 0-2] at 5 min). The group receiving moxifloxacin 0.5% showed an increase in conjunctival vascularity (0.55 [range: 0-1] at baseline to 1.61 [range: 0.5-3] at 5 min.) that was significantly greater (p = 0.0005) compared with that of the group receiving gatifloxacin 0.3% (0.52 [range: 0-1] at baseline to 0.68 [range: 0-2] at 5 min.). Significantly less pain (1.2 vs. 3.2, p = 0.001) and irritation (0.64 vs. 3.42, p = 0.001) occurred with gatifloxacin 0.3% than with moxifloxacin 0.5%. Pupil size was significantly reduced (5.65 mm-5.05 mm) in eyes receiving moxifloxacin 0.5% (p = 0.004) and no significant change occurred in pupil size (5.60 mm-5.65 mm) in eyes that received gatifloxacin 0.3% (p = 0.878). No AC reaction was noted with either medication. CONCLUSIONS: The group of eyes receiving gatifloxacin 0.3% with BAK demonstrated greater ocular tolerability in comparison to the group receiving moxifloxacin 0.5% without BAK. Moxifloxacin induced pupillary miosis may be due to prostaglandin release in the anterior chamber. A limitation of this study is the relatively young age of the study population. PMID- 15537476 TI - Effects of tirofiban on acute systemic inflammatory response in elective percutaneous coronary interventions. AB - OBJECTIVE: In this study the effect of a specific glycoprotein IIb/IIIa inhibitor, tirofiban [which also has antiplatelet activity on acute systemic inflammatory responses (IR) during elective percutaneous coronary intervention (PCI)] was evaluated. PATIENTS AND METHODS: Patients with stable angina pectoris and similar baseline characteristics who angiographically had a single lesion in their coronary arteries with a PCI performed on that lesion were enrolled in the study. One group of patients (control group, n = 52) received 0.9% NaCl (15 mL/h for 24 h) and the other group (tirofiban group, n = 55) had tirofiban (10 microg/kg bolus infusion in 3 min and 0.15 microg/kg/min for 24 h) in addition to stenting without pre-dilatation. The effect of interventional procedure on levels of cardiac troponin T (cTnT) and several parameters of acute IR (leukocytes, fibrinogen, C-reactive protein, interleukin-1, interleukin-6, interleukin-8 and tumor necrotizing factor-alpha) was assessed on blood samples obtained from all patients before PCI and at pre-specified time points after PCI. RESULTS: During the follow-up after PCI, the number of patients becoming cTnT-positive (> 0.1 ng/mL) was greater in the control group [12 (23%) patients vs. 3 (5%) patients, p = 0.01]. However, both groups had changes (generally observed as elevations) in their levels of all inflammatory parameters during the study and C-reactive protein, interleukin-6 and tumor necrotizing factor-alpha levels were elevated significantly. Yet, no significant difference occurred between groups due to these changes in any phase of the study (p > 0.05). CONCLUSIONS: Based on the findings of this study, it was concluded that although tirofiban limits development of myocardial necrosis during elective PCI, it does not directly affect the acute systemic inflammatory responses. PMID- 15537477 TI - How to improve current therapeutic standards in upper respiratory infections: value of fusafungine. AB - Despite guidelines and educational programs, systemic antibiotics and anti inflammatory drugs are often inappropriately prescribed in upper respiratory tract infections (URTIs), although they are most often of viral origin, generally benign, and self-limiting with spontaneous recovery in more than 80% of cases. Reduced use of systemic antibiotics is crucial in the current context of concern about emerging antibiotic resistance and reducing unnecessary costs associated both with drug over-consumption and with the management of the consequences of antibiotic resistance. Local bacterial or viral infection of the airways induces an early inflammatory reaction. Although this inflammatory reaction has a beneficial effect in the capture and destruction of the pathogens, it can be responsible for deleterious tissue damage and vascular alterations leading to a self-perpetuating cycle of events. A wide array of medicines is available for symptomatic relief of URTIs: many of them are partially effective in reducing symptoms, but none is curative. Local administration of antibiotics and anti inflammatory drugs allows drug delivery directly to the target site of infection and inflammation, i.e., the respiratory mucosa, thus enabling a higher concentration of the drug, which results in smaller doses to be given, decreased potential for systemic toxicity, fewer side effects, protection of other flora, and rapid relief. Fusafungine is a naturally occurring peptide antibiotic with anti-inflammatory properties, which selectively targets the tissue reaction and preserves the natural antibacterial and antiviral defences. It is indicated for topical use in nose and throat infections. A recent analysis of French general practitioners' (GPs) prescribing pattern in the field of URTIs has demonstrated that prescription of fusafungine has achieved what many educational programs have failed to do: a significant reduction in the 'real life' prescription of systemic antibiotics and antiinflammatory drugs, without the side effects of corticosteroids and vasoconstrictive agents, and without impact on microbial ecology. PMID- 15537478 TI - Migraine treatment strategies: the global Migraine And Zolmitriptan Evaluation (MAZE) survey--phase IV. AB - OBJECTIVE: To determine which medications UK migraineurs have access to and assess the usage of these products in a 'real-life' setting. RESEARCH DESIGN AND METHODS: Data were collected using an online questionnaire, which subjects were directed to by advertisements in pharmacies, a UK national newspaper, on the internet and information on the Migraine Action Association website and newsletters. Respondents were eligible for inclusion if attacks fulfilled International Headache Society criteria for migraine and/or if previously diagnosed by a physician as having migraine. MAIN OUTCOME MEASURES: Respondents were asked to specify which products had been prescribed or purchased for migraine treatment. The pattern of use of these products was determined, including the reasons why respondents chose particular products to treat attacks. RESULTS: Of 3072 eligible respondents, the majority had purchased a variety of over-the-counter (OTC) medicines for treatment of attacks. Eighty-seven per cent had been diagnosed by a physician and were prescribed multiple products (average 1.68); 45% received triptans, but 26% were still prescribed products that were also available OTC. Over half (52%) of respondents initially used an OTC medicine to treat the last migraine attack. However, 73% required a second dose/product, mainly as a result of lack of efficacy of the first dose/product. Respondents using triptans were less likely to require a second dose/product than those not using triptans (52% vs. 78%, respectively). The two main reasons for choosing a triptan to treat an attack were the need for quick control and the severity of the attack. Satisfaction with regard to migraine medication was higher among triptan-users than nontriptan users. CONCLUSIONS: Medicines that are available OTC are often used as first-line therapy for migraine despite many migraineurs having access to prescription therapies such as triptans. Many migraineurs require a second dose/product, possibly indicating sub-optimal treatment efficacy. Physicians should consider the range of migraine-specific treatments available, including triptans, in order to develop a treatment plan that is based on the patient's needs and preferences. PMID- 15537479 TI - Diagnosing restless legs syndrome (RLS) in primary care. AB - This paper represents a review of current opinion and information on the effective diagnosis of restless legs syndrome (RLS) in a primary care setting. RLS can be a distressing condition--it can cause serious sleep disturbance and has a significant impact on quality of life comparable to that of depression or type 2 diabetes. The prevalence of adults whose RLS is severe enough to warrant medical advice has been estimated to be approximately 3%, but only a small proportion of these patients currently report having been diagnosed in primary care, despite stating that they have presented to their GP. The benefits of increased understanding of the symptoms of RLS and how patients present in primary care are discussed, with emphasis on how this will help GPs more effectively diagnose and manage the patients affected. Guidelines on how to diagnose RLS in a primary care setting are given--when a patient presents with sleep disturbance, RLS should be routinely considered and, where existing, be readily diagnosed in a primary care setting on the basis of the patient's clinical history, a physical examination and with the aid of four questions based on the International RLS Study Group (IRLSSG) four essential diagnostic criteria. PMID- 15537480 TI - The role of angiotensin antagonism in stroke prevention in patients with hypertension: focus on losartan. AB - Optimal management of hypertension has been shown to reduce the risk of stroke. In recent years, newer classes of antihypertensive such as the angiotensin II (Ang II) antagonists have become available. Results from the Losartan Intervention for Endpoint reduction in hypertension (LIFE) study suggest the utility of this particular Ang II antagonist in stroke prevention. Treatment with a losartan-based regimen or an atenolol-based regimen produced similar reductions in blood pressure during almost 5 years of follow up. Losartan, however, reduced the risk of stroke by 25% compared with atenolol (p = 0.001). For a subgroup of patients with isolated systolic hypertension, losartan reduced the risk of stroke by 40% (p = 0.02). As well as blocking the Ang II type 1 receptor, losartan also acts as an antagonist at the thromboxane A2 receptor and has uricosuric effects, which may provide additional mechanisms by which losartan provides protective benefits beyond its antihypertensive action. The relevance of these molecular properties of losartan over other Ang II antagonists is further supported by comparison of the outcomes obtained in clinical trials employing two other Ang II antagonists, valsartan and candesartan. PMID- 15537481 TI - The impact of a worksite migraine intervention program on work productivity, productivity costs, and non-workplace impairment among Spanish postal service employees from an employer perspective. AB - BACKGROUND: Migraine is associated with a significant productivity loss to employers, who may benefit from making a migraine intervention available to their employees. OBJECTIVE: To evaluate changes in migraine-related productivity and non-workplace impairment associated with a migraine intervention program from the employer perspective. METHODS: This was a pre-test post-test study of Spanish Postal Service employees with migraine. The intervention consisted of counseling from occupational health physicians and rizatriptan 10 mg for symptomatic treatment of two subsequent migraine headaches. Physicians also prescribed additional medications for migraine prophylaxis, treatment of tension headaches, and rescue medications. Migraine-related work loss and non-workplace impairment (interference with daily and social activities) were self-reported at baseline (pre-intervention) and separately following each migraine headache (post intervention) with the aid of a diary. Migraine-related work loss was reported as work loss due to absenteeism, reduced productivity while at work, and the sum of the two (total lost work day equivalents [LWDE]). An employer perspective was taken for the cost analysis, and thus productivity costs were the only costs considered. RESULTS: A total of 436 patients comprised the population for analysis. The number of migraine-related LWDE per migraine attack were 0.48 days per migraine headache in the month before the intervention, decreasing to 0.20 days and 0.07 days per migraine headache during the first and second migraine headaches following the intervention (p < 0.0001 vs. baseline). Total migraine related productivity costs per migraine headache were 34 euros/patient before the intervention, decreasing to 14 euros/patient and 5 euros/patient during the first and second headaches following the intervention (p < 0.0001). Non-workplace activity impairment was also significantly reduced (p < 0.0001) following the intervention. The main limitations of the study were the lack of a parallel control group and the potential for differential recall bias between the usual care and the intervention periods. In addition, the results may not be generalizable to other employers or other countries. CONCLUSION: This study documents the value of a workplace migraine intervention program, which focused on migraine prevention and rizatriptan-based symptomatic treatment. It also highlights the important role occupational health clinics can play in helping employers and employees reduce the burden of migraine. PMID- 15537482 TI - Effects of galantamine in patients with mild Alzheimer's disease. AB - BACKGROUND: Galantamine is an acetylcholinesterase inhibitor that modulates nicotinic receptors. It is effective in mild to moderate Alzheimer's disease (AD) but no trial has focused exclusively on mild AD. We performed a post-hoc sub-set analysis using data from four randomised trials to explore the efficacy of galantamine versus placebo in mild AD. METHODS: Participants in all studies met NINCDS-ADRDA criteria for probable AD. We examined data from patients with baseline Mini Mental State Examination (MMSE) 21-24 who received galantamine 24 mg/day (GAL) or placebo (PLAC). Scores for the Alzheimer's Disease Assessment Scale-cognitive subset (ADAS-cog), Clinician's Interview-Based Impression of Change (CIBIC), Disability Assessment for Dementia (DAD), and ACDS-ADL scales were compared. RESULTS: Of the 694 patients (362 GAL, 332 PLAC, mean baseline MMSE 22.4 +/- 1.1, mean age 74 +/- 7.9 years), 65% completed 6 months treatment (223 GAL, 229 PLAC). Mean change in ADAS-cog at 6 months was -1.5 (95% confidence interval -2.2, -0.8, p < 0.001) for GAL and +0.2 (-0.6, 0.9, p = 0.72) for PLAC. This difference was statistically significant (p = 0.001). Significantly more patients receiving galantamine were classified as 'improved' using the CIBIC (26.9% GAL vs 14.3% PLAC, p < 0.001). Galantamine was generally well tolerated; most common adverse events were nausea, vomiting and diarrhoea. CONCLUSIONS: Pooled data from four randomised trials of patients with mild AD indicate that patients who received galantamine 24 mg/day for 6 months improved cognition more often than those who received placebo and that a higher proportion receiving galantamine were globally improved. This suggests that patients with mild AD benefit from galantamine treatment. PMID- 15537483 TI - Clopidogrel and vascular disease prevention. AB - In this editorial we consider some aspects of the use of clopidogrel in current clinical practice. Clopidogrel has replaced ticlopidine as an alternative to aspirin. Among the problems with the use of antiplatelet agents is aspirin intolerance. The options for these patients are discussed; these include using clopidogrel. Clopidogrel may be especially effective in patients with peripheral arterial disease. However, this impression requires confirmation in appropriately designed trials. We also consider the effect of antiplatelet agents on growth factor release by platelets and of clopidogrel on the inflammatory process. Clopidogrel is a useful, evidence-based, antiplatelet agent. PMID- 15537484 TI - Role of antiplatelet therapy in cardiovascular disease I: Acute coronary syndromes. AB - The acute coronary syndromes (ACS), consisting of ST-segment elevation myocardial infarction (STEMI), non-STEMI (NSTEMI), and unstable angina, remain a leading cause of death in the United States. Through the process of atherothrombosis, underlying atherosclerosis can progress to an acute ischemic coronary event. This disease mechanism is also common to ischemic stroke and peripheral arterial disease. As ACS is a heterogeneous disease, accurate patient diagnosis and risk categorization is essential. Treatment approaches for both STEMI and NSTEMI ACS consist of a combination of surgical intervention and pharmacotherapy, with antiplatelet agents such as clopidogrel, aspirin and glycoprotein IIb/IIIa receptor antagonists playing an essential role. PMID- 15537485 TI - Role of antiplatelet therapy in cardiovascular disease II: Ischemic stroke. AB - The etiology of cerebrovascular disease is heterogeneous, with the majority of strokes being of ischemic origin. Transient ischemic attack is now considered to be an important precursor and long-term risk factor for ischemic stroke. Given the lack of acute therapies for ischemic stroke, current treatments focus on secondary prevention through risk-factor management, pharmacotherapy and interventional approaches. As illustrated in this paper, antiplatelet agents (e.g. clopidogrel, aspirin, dipyridamole) are the cornerstone of therapy for prevention of recurrent ischemic stroke. PMID- 15537486 TI - Role of antiplatelet therapy in cardiovascular disease III: Peripheral arterial disease. AB - Peripheral arterial disease (PAD) is a common manifestation of the atherosclerotic disease process, typically affecting vascular beds in the lower extremities. In its most severe form PAD may lead to limb amputation. Patients with PAD are also at increased cross-risk of thrombosis at coronary and cerebrovascular sites. However, despite its prevalence and severity, PAD is underdiagnosed and undertreated. In this collection of case studies, the role of the antiplatelet agent clopidogrel in current treatment strategies for the management of PAD is highlighted. PMID- 15537487 TI - Focal adhesion kinase and angiogenesis. Where do we go from here? PMID- 15537488 TI - Nitric oxide and efficiency of the right heart. PMID- 15537489 TI - The neuroendocrine-immune interface gone awry in aldosteronism. PMID- 15537490 TI - The fine-scale architecture of defibrillation. PMID- 15537491 TI - Identifying and understanding the role of pulmonary vein activity in atrial fibrillation. AB - The perception of atrial fibrillation development has changed drastically over the last decade. The pulmonary veins have been targeted as the source of arrhythmogenic activity involved in the initiation of atrial fibrillation. This activity appears to be localized in the myocardial sleeves of the vessels. Extensive study of cells within this tissue has helped create a new model for atrial fibrillation. This review attempts to show how the development, architecture and electrophysiologic properties of the pulmonary veins influence the initiation and perpetuation of atrial fibrillation. It also examines the potential long-term effects of pulmonary vein activity on arrhythmia development. PMID- 15537492 TI - Therapeutic opportunities for cell cycle re-entry and cardiac regeneration. AB - Over the last two decades, considerable effort has been made to better understand putative regulators and molecular switches that govern the cell cycle in attempts to reactivate cell cycle progression of cardiac muscle. Rapid advancements on the field of stem cycle biology including evidence of cardiac progenitors within the adult myocardium itself and reports of cardiomyocyte DNA synthesis, which each suggest that the adult myocardium may in fact have the capacity for de novo myocyte regeneration. Augmenting cardiomyocyte number by targeting specific cell cycle regulatory genes or by stimulating cardiac progenitor cells to differentiate into cardiac muscle may be of therapeutic value in repopulating the adult myocardium with functionally active cells in patients with end-stage heart failure. Advancements in the area of cardiomyocyte cell cycle control and regeneration and their therapeutic potential are discussed. PMID- 15537493 TI - Transcriptional regulation of the murine Connexin40 promoter by cardiac factors Nkx2-5, GATA4 and Tbx5. AB - OBJECTIVE: Connexin40 (Cx40) is a gap junction protein expressed specifically in developing and mature atrial myocytes and cells of the conduction system. In this report, we identify cis-acting elements within the mouse Cx40 promoter and unravel part of the complex pathways involved in the cardiac expression of this gene. METHODS: To identify the factors involved in the cardiac expression of Cx40, we used transient transfections in mammalian cells coupled with electrophoretic mobility shift assays (EMSA) and RT-PCR. RESULTS: Within the promoter region, we identified the minimal elements required for transcriptional activity within 150 base pairs (bp) upstream of the transcriptional start site. Several putative regulatory sites for transcription factors were predicted within this region by computer analysis, and we demonstrated that the nuclear factors Sp1, Nkx2-5, GATA4 and Tbx5 could interact specifically with elements present in the minimal promoter region of the Cx40. Furthermore, co-transfection experiments showed the ability of Nkx2-5 and GATA4 to transactivate the minimal Cx40 promoter while Tbx5 repressed Nkx2-5/GATA4-mediated activation. Mutagenesis of the Nkx2-5 core site in the Cx40 promoter led to significantly decreased activity in rat smooth muscle cell line A7r5. Consistent with this, mouse embryos lacking Nkx2-5 showed a marked decrease in Cx40 expression. CONCLUSION: In this work, we cloned the promoter region of the Cx40 and demonstrated that the core promoter was modulated by cardiac transcriptional factors Nkx2-5, Tbx5 and GATA4 acting together with ubiquitous Sp1. PMID- 15537494 TI - HMG-CoA reductase inhibitor attenuates experimental autoimmune myocarditis through inhibition of T cell activation. AB - OBJECTIVE: This study tested the hypothesis that 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibitor affects T cell-mediated autoimmunity through inhibition of nuclear factor-kappaB (NFkappaB) and reduces the severity of experimental autoimmune myocarditis (EAM). METHODS: EAM was induced in Lewis rats by immunization with myosin. High-dose or low-dose fluvastatin or vehicle was administered orally for 3 weeks to rats with EAM. RESULTS: Fluvastatin reduced the pathophysiological severity of myocarditis. Fluvastatin inhibited expression of NFkappaB in the nuclei of myocardium in EAM. Fluvastatin reduced production of Th1-type cytokines, including interferon (IFN)-gamma and interleukin (IL)-2, and inhibited expression of inflammatory cytokine mRNAs in the myocardium. Infiltration of CD4-positive T cells into the myocardium and T cell proliferative responses were suppressed by fluvastatin. Plasma lipid levels did not differ between the groups. CONCLUSIONS: Fluvastatin ameliorates EAM by inhibiting T cell responses and suppressing Th1-type and inflammatory cytokines via inactivation of nuclear factor-kappaB, and this activity is independent of cholesterol reduction. PMID- 15537495 TI - Overexpression of focal adhesion kinase in vascular endothelial cells promotes angiogenesis in transgenic mice. AB - OBJECTIVE: Focal adhesion kinase is implicated in the regulation of cell adhesion, migration, survival, and cell-cycle progression. However, the functions of focal adhesion kinase in endothelial cell (EC) in vivo remain unclear. This study aims to examine the role of FAK in EC function and angiogenesis in vivo by transgenic mice approach. METHOD: We generated transgenic mice which overexpressed chicken FAK in vascular endothelial cell under the control of the Tie-2 promoter and enhancer. FAK transgene was detected by RT-PCR, immunoprecipitation, and Western blot. The effect of FAK overexpression on angiogenesis was determined using skin wound healing and ischemia skeleton muscle models. RESULTS: Expression of FAK transgene was detected in all vessel-rich tissues. Expression of FAK protein was verified by antibody specific for the exogenous chicken FAK in lung homogenates and isolated EC. In the wound-induced angiogenesis model, the number of vessels in the granulation tissue of healing wound was significantly increased in the transgenic mouse compared to that of wild-type control mice. Similarly, in the ischemia skeleton muscle model, the density of capillaries was significantly increased in the transgenic mouse. CONCLUSION: These results indicate that FAK may play an important role in the promotion of angiogenesis in vivo. PMID- 15537496 TI - Nitric oxide contributes to oxygen demand-supply balance in hypoperfused right ventricle. AB - OBJECTIVE: The present study examined the role of nitric oxide (NO) in oxygen demand-supply balance in hypoperfused canine right ventricular myocardium. METHODS: The right coronary artery of anesthetized, open-chest dogs was perfused at pressures of 80, 60, and 40 mm Hg, and right ventricular myocardial oxygen consumption, right coronary blood flow and other hemodynamic and cardiac function variables were measured. Right ventricular mechanical function was indexed as the product of heart rate x peak right ventricular systolic pressure x right ventricular dP/dt(max). NO synthesis blocker N(omega)-nitro-L-arginine methyl ester (L-NAME, 150 mug/min) was infused into the right coronary artery to block NO synthesis. RESULTS: Neither hypoperfusion nor L-NAME altered right ventricular function. Right ventricular myocardial oxygen consumption fell with coronary perfusion pressure, but less steeply after L-NAME, and at all perfusion pressures was elevated above control. The increase in myocardial oxygen consumption in the absence of NO was met by increased oxygen extraction and by non-NO dependent vasodilation, but the relationship between flow and oxygen consumption was displaced downward after L-NAME. As right coronary perfusion pressure was reduced, the relationship between right ventricular oxygen consumption and right coronary venous PO(2) became steeper after L-NAME, and right coronary venous PO(2) was significantly reduced. CONCLUSIONS: During right coronary hypoperfusion, right ventricular function is well maintained, but myocardial oxygen consumption falls, reflecting an increase in oxygen utilization efficiency. NO contributes to this adaptation to hypoperfusion by restraining myocardial oxygen consumption, and by promoting coronary vasodilation with less severe reduction in myocardial PO(2). NO has an important role in right ventricular oxygen demand-supply balance when right coronary perfusion pressure is reduced. PMID- 15537497 TI - Critical role of CNS effects of aldosterone in cardiac remodeling post-myocardial infarction in rats. AB - BACKGROUND: Oral administration of spironolactone improves cardiac remodeling and its central infusion prevents the increase in sympathetic drive post-myocardial infarction (MI). We hypothesized that central actions of aldosterone contribute to cardiac remodeling post-MI. OBJECTIVE: To compare the effects of intracerebroventricular (icv) infusion and oral administration of spironolactone on cardiac remodeling and left ventricle (LV) dysfunction post-MI in rats. METHODS: Spironolactone was administered orally (80 mg/kg/day) or by icv infusion (100 ng/h), starting 1-3 days post-MI in Wistar rats and continued for 6 weeks. RESULTS: At 6 weeks post-MI, in the rats treated with vehicle, LV peak systolic pressure (LVPSP) and LV dP/dt max were clearly decreased and LV end diastolic pressure (LVEDP) and plasma catecholamines and serum aldosterone increased. All these parameters improved with both oral and icv spironolactone. The MI-induced increases in internal circumferences of LV and right ventricle (RV), and in interstitial and perivascular fibrosis, in both the LV and RV were significantly prevented/inhibited by both oral and icv spironolactone. Laminin, fibronectin and fibrillar collagen (visualized by scanning electron microscopy, SEM) increased in the non-infarcted part of the LV post-MI in the vehicle group, but not/less in rats on oral or icv spironolactone. CONCLUSIONS: Since the magnitude of beneficial effects of icv spironolactone at low doses was largely equal to that achieved with its oral administration at much higher doses, we propose that in addition to other sites of action, aldosterone appears to activate central nervous system (CNS) pathways and thereby influences peripheral mechanisms involved in cardiac remodeling. PMID- 15537498 TI - High-resolution optical mapping of intramural virtual electrodes in porcine left ventricular wall. AB - OBJECTIVE: It is believed that shock-induced intramural virtual electrodes (IVE) play a critical role in defibrillation. IVE were recently demonstrated in the porcine left ventricle (LV), but their origin remains unknown. Macroscopic optical mapping showed that strong shocks induce IVE of only one polarity, which contradicts theoretical predictions. It is hypothesized that IVE have a microscopic origin and that microscopic positive and negative IVE are spatially averaged during macroscopic optical mapping. This hypothesis was examined by mapping V(m) responses at the transmural LV surface with increased optical resolution. METHODS: Rectangular shocks (strength=2-48 V/cm; duration=10 ms) were applied across isolated coronary-perfused porcine LV preparations (n=7) during the action potential plateau and diastole. Shock-induced V(m) responses were measured at low resolution (LR; 1.2 mm/diode) and high resolution (HR; 0.11 mm/diode). RESULTS: During plateau shocks with strength > or =20 V/cm, LR recordings demonstrated only negative DeltaV(m) extending to the cathodal preparation edge. In contrast, HR recordings from this area as well as from intramural locations revealed both positive and negative DeltaV(m) at all shock strengths. During diastolic shocks, only positive polarizations were observed at LR, but both positive and negative polarizations were detected at HR. In areas of negative polarization, large activation delays were found at HR, whereas LR recordings at these locations demonstrated fast activation. CONCLUSIONS: High- and low-resolution optical mapping produced radically different patterns of shock induced polarization and activation. The occurrence of positive and negative polarizations during plateau and diastolic shocks at high but not low resolution provides evidence for microscopic nature of IVE in LV wall. PMID- 15537499 TI - Diltiazem inhibits hKv1.5 and Kv4.3 currents at therapeutic concentrations. AB - OBJECTIVE: In the present study we examined the effects of diltiazem, an L-type Ca(2+) channel blocker widely used for the control of the ventricular rate in patients with supraventricular arrhythmias, on hKv1.5 and Kv4.3 channels that generate the cardiac ultrarapid delayed rectifier (I(Kur)) and the 4 aminopyridine sensitive transient outward (I(to)) K(+) currents, respectively. METHODS: hKv1.5 and Kv4.3 channels were stably and transiently expressed in mouse fibroblast and Chinese hamster ovary cells, respectively. Currents were recorded using the whole-cell patch clamp. RESULTS: Diltiazem (0.01 nM-500 muM) blocked hKv1.5 channels, in a frequency-dependent manner exhibiting a biphasic dose response curve (IC(50)=4.8+/-1.5 nM and 42.3+/-3.6 muM). Diltiazem delayed the initial phase of the tail current decline and shifted the midpoint of the activation (Vh=-16.5+/-2.1 mV vs -20.4+/-2.6 mV, P<0.001) and inactivation (Vh= 22.4+/-0.7 mV vs. -28.2+/-1.9 mV, P<0.001) curves to more negative potentials. The analysis of the development of the diltiazem-induced block yielded apparent association (k) and dissociation (P) rate constants of (1.6+/-0.2) x 10(6) M( 1)s(-1) and 46.8+/-4.8 s(-1), respectively. Diltiazem (0.1 nM-100 muM) also blocked Kv4.3 channels in a frequency-dependent manner exhibiting a biphasic dose response curve (IC(50)=62.6+/-11.1 nM and 109.9+/-12.8 muM). Diltiazem decreased the peak current and, at concentrations > or =0.1 microM, accelerated the inactivation time course. The apparent association and dissociation rate constants resulted (1.7+/-0.2) x 10(6) M(-1)s(-1) and 258.6+/-38.1 s(-1), respectively. Diltiazem, 10 nM, shifted to more negative potentials the voltage dependence of Kv4.3 channel inactivation (Vh=-33.1+/-2.3 mV vs -38.2+/-3.5 mV, n=6, Plt;0.05) the blockade increasing at potentials at which the amount of inactivated channels increased. CONCLUSION: The results demonstrated for the first time that diltiazem, at therapeutic concentrations, decreased hKv1.5 and Kv4.3 currents by binding to the open and the inactivated state of the channels. PMID- 15537500 TI - Direct block of hERG potassium channels by the protein kinase C inhibitor bisindolylmaleimide I (GF109203X). AB - OBJECTIVE: The human ether-a-go-go-related gene (hERG) encodes the rapid component of the cardiac repolarizing delayed rectifier potassium current, I(Kr). The direct interaction of the commonly used protein kinase C (PKC) inhibitor bisindolylmaleimide I (BIM I) with hERG, KvLQT1/minK, and I(Kr) currents was investigated in this study. METHODS: hERG and KvLQT1/minK channels were heterologously expressed in Xenopus laevis oocytes, and currents were measured using the two-microelectrode voltage clamp technique. In addition, hERG currents in stably transfected human embryonic kidney (HEK 293) cells, native I(Kr) currents and action potentials in isolated guinea pig ventricular cardiomyocytes were recorded using whole-cell patch clamp electrophysiology. RESULTS: Bisindolylmaleimide I blocked hERG currents in HEK 293 cells and Xenopus oocytes in a concentration-dependent manner with IC(50) values of 1.0 and 13.2 muM, respectively. hERG channels were primarily blocked in the open state in a frequency-independent manner. Analysis of the voltage-dependence of block revealed a reduction of inhibition at positive membrane potentials. BIM I caused a shift of -20.3 mV in the voltage-dependence of inactivation. The point mutations tyrosine 652 alanine (Y652A) and phenylalanine 656 alanine (F656A) attenuated hERG current blockade, indicating that BIM I binds to a common drug receptor within the pore region. KvLQT1/minK currents were not significantly altered by BIM I. Finally, 1 muM BIM I reduced native I(Kr) currents by 69.2% and lead to action potential prolongation. CONCLUSION: In summary, PKC-independent effects have to be carefully considered when using BIM I as PKC inhibitor in experimental models involving hERG channels and I(Kr) currents. PMID- 15537501 TI - Divergent action potential morphologies reveal nonequilibrium properties of human cardiac Na channels. AB - OBJECTIVE: Fast inward Na current (I(Na)) carried by the voltage-gated Na channel (Na(V)1.5) is critical for action potential (AP) propagation and the rapid upstroke of the cardiac AP. In addition, a small fraction of Na(V)1.5 channels remains open throughout the plateau of the AP, and this current is termed as late I(Na). In patients with mutant Na(V)1.5-based congenital long Q-T (LQT) syndrome, mutant channels pass more late I(Na) compared to wild-type channels in unaffected patients. Although LQT mutant Na(V)1.5 channels are well studied, there is no careful evaluation of the effects of cardiac APs on early and late current. This is important with the recent documentation of nonequilibrium I(Na). METHODS: We measured AP-stimulated I(Na) through Na(V)1.5 wild-type and two LQT mutant channels (DeltaKPQ and N1325S). Three distinct AP morphologies were used: human embryonic stem cell-derived cardiac myocyte (hES-CM) APs with a relatively slow upstroke and canine endocardial and epicardial ventricular myocytes with rapid upstrokes. RESULTS: All three APs elicited both early and late I(Na). For wild type Na(V)1.5, the hES-CM AP elicits more early and late I(Na) than either the endocardial or epicardial AP. The mechanism for this difference is that the hES CM has a relative slow dV/dt(max) that causes a maximal open channel probability. Slower upstroke stimulation also allows greater Na flux through wild-type and N1325S channels, but not the DeltaKPQ mutant. CONCLUSIONS: The inherent gating properties of Na(V)1.5 provide natural tuning of optimal I(Na) density. Slower upstroke velocities can yield more I(Na) and Na flux in some Na(V)1.5 variants. PMID- 15537502 TI - Heart block in mice overexpressing calcineurin but not NF-AT3. AB - OBJECTIVE: Overexpression of calcineurin causes cardiac hypertrophy and arrhythmic deaths. During disease development, sinus bradycardia followed by high degree atrioventricular (AV) block finally culminating in ventricular asystole has been observed over time in calcineurin hearts. AV block is associated with the development of pleomorphic ventricular tachycardia in mice and downregulation of potassium currents in ventricular myocytes. We tested the hypothesis that the abnormalities of AV block and propensity to ventricular tachycardia relate to overexpression of calcineurin independent of the development of hypertrophy. METHODS: Cardiac electrophysiologic properties were compared in isolated perfused hearts with ventricular hypertrophy due to overexpression of calcineurin or NF AT3 and in their corresponding wild types at 15 or 30 days of age. RESULTS: Compared to wild-type hearts, significant prolongation of sinus node recovery times was noted in both NF-AT3 and calcineurin hearts. Compared to wild-type hearts, Wenckebach cycle length (WCL) and the left ventricular effective refractory period (LVERP) were significantly prolonged in the calcineurin hearts (p<0.05) but not NF-AT3 hearts. In calcineurin mice, left ventricular effective refractory period impinged on Wenckebach cycle length resulting in a significant correlation between left ventricular effective refractory period and Wenckebach cycle length (r(2)=0.56). No such correlation was observed for wild type or NF AT3 hearts. At 30 days of development, ventricular tachycardia developed in 70% of calcineurin hearts compared to 0% wild-type hearts (p=0.003), whereas ventricular tachycardia was observed in 33% of NF-AT3 hearts and 10% of corresponding wild-type hearts (p=NS). CONCLUSIONS: The prolonged ventricular refractoriness, seen only in calcineurin hearts, impinges on Wenckebach cycle length resulting in heart block and is associated with propensity to ventricular tachycardia. PMID- 15537503 TI - Externalization of endogenous annexin A5 participates in apoptosis of rat cardiomyocytes. AB - OBJECTIVE: Annexins are Ca(2+)-dependent phospholipid binding proteins. Externalized annexin A5 has been recently suggested to have a proapoptotic effect. Our aim was to determine whether annexin A5, which is intracellular in cardiomyocytes, could be translocated and/or externalized and play a role during the apoptotic process. METHODS: Apoptosis was induced in rat cardiomyocytes by continuous incubation with staurosporine or 30 min treatment with H(2)O(2) and was measured by phosphatidylserine (PS) externalization, TUNEL staining and DNA ladder. Immunofluorescence labeling of annexin A5 was performed on permeabilized or nonpermeabilized cardiomyocytes. RESULTS: Staurosporine or H(2)O(2) treatment of neonatal cardiomyocytes resulted in significant increases of apoptosis at 24 h, but H(2)O(2) treatment led to a faster and higher PS externalization than that observed with ST. In both neonatal and adult cardiomyocytes, annexin A5 was intracellular in control conditions but was found at the external face of sarcolemma during apoptosis. Furthermore, neonatal cardiomyocytes with externalized annexin A5 have apoptotic characteristics and their number increased with time. Interestingly, immediately after H(2)O(2) induction, the number of annexin A5-positive cells was higher than that of PS-positive cells (p1,200 patients enrolled. Myth no. 3--Improvement in clinical trials of cholinergics in VaD is due to underlying AD, not to the vascular lesions. Experimental, clinical and pathological evidence has demonstrated cholinesterase deficits in VaD (independently of any concomitant AD pathology), including low acetylcholine in cerebrospinal fluid, and reduced choline acetyltransferase (ChAT) in the brain. PMID- 15537520 TI - Is poststroke depression a vascular depression? AB - As we learn more about the relationships between depression and cerebrovascular disease (CVD), a complex picture is emerging in which the chain of causality seems to spiral on itself: progressive or focal brain damage, cognitive impairment, depressive symptoms, dementia, and cardiovascular diseases, all seem to be liable to lead to one or another. Stroke may lead to depression, and the inverse may also be true. Depression may lead to cognitive impairment and cardiovascular diseases, which in turn may lead to subtle brain impairment, thereby causing more depression and cognitive impairments, and so on. In this presentation, we provide a rapid glance at the complexities of such issues. PMID- 15537521 TI - Establishing diagnostic criteria for vascular depression. AB - We have proposed a subtype of vascular depression with two key elements: (1) presence of major depression and (2) presence of cerebrovascular disease on neuroimaging. Future studies are needed to refine these diagnostic criteria. There must be a consensus as to whether requiring major depression criteria is acceptable, or possibly too stringent. There are several methodological issues regarding cerebrovascular disease, including defining severity and thresholds, importance of location and system of assessment. Once agreement is reached on a working diagnosis of vascular depression, researchers will be in a position to estimate prevalence of the disorder, determine conditions that represent risk factors, and examine important outcomes in patients with vascular depression. PMID- 15537522 TI - Treatment of vascular dementia--evidence from clinical trials with cholinesterase inhibitors. AB - Cerebrovascular disease (CVD), as well as secondary ischemic brain injury from cardiovascular disease, are common causes of dementia and cognitive decline in the elderly. In addition, CVD frequently contributes to cognitive loss in patients with Alzheimer's disease (AD). Progress in understanding the pathogenetic mechanism involved in vascular cognitive impairment (VCI) and vascular dementia (VaD) has resulted in promising treatments of these conditions. Cholinergic deficits in VaD are due to ischemia of basal forebrain nuclei and of cholinergic pathways and can be treated with the use of the cholinesterase inhibitor agents used in AD. Controlled clinical trials with donepezil, galantamine and rivastigmine in VaD, as well as in patients with AD plus CVD, have demonstrated improvements in cognition, behavior and activities of daily living. PMID- 15537523 TI - Treatment of vascular dementia: evidence from trials with non-cholinergic drugs. AB - A considerable number of therapeutic trials have been performed in vascular dementia (VaD). The results of these trials have generally been considered as disappointing and no drug treatment has been so far approved for the treatment of VaD by regulatory agencies. The aim of the present paper is to critically review the results of randomized clinical trials performed with non-cholinergic drugs in VaD. The conclusions of the present review are that: (1) some drugs such as nicergoline, memantine, posatirelin, propentofylline, and pentoxifylline have shown some, although partly limited, benefits in VaD patients; (2) besides a lack of efficacy of the tested drugs, possible causes of the negative results of many randomized clinical trials in VaD are the enrollment of patients with heterogeneous subtypes of VaD, the small sample size, and the use of end-points and cognitive tests inadequate for the VaD setting because derived from previous experience in the field of Alzheimer disease. Preliminary analyses show that focusing therapeutic trials on specific forms of VaD such as the subcortical type may lead to results different from those obtained in a heterogeneous VaD group. This selective focus seems to be better suited for disclosing specific treatments in the field of VaD. PMID- 15537524 TI - Prevention of dementia: lessons from SYST-EUR and PROGRESS. AB - Hypertension is one of the principal risk factors for cerebrovascular diseases, closely correlated also with cognitive decline and dementia. Data from recent therapeutic trials (SYST-EUR, PROGRESS) open the way toward the prevention of dementia (vascular or Alzheimer's type) by antihypertensive treatments. The results of these two studies suggest different mechanisms of action of antihypertensive drugs in the prevention of cognitive decline. The use of angiotensin converting enzyme (ACE) inhibitors, with or without diuretics, resulted in decrease incidence of stroke-related dementia, but dementia without stroke was not reduced. With the dihydropyridine calcium antagonists, a reduction in both Alzheimer's type and vascular dementia was demonstrated. PMID- 15537525 TI - Towards defining the neuropathological substrates of vascular dementia. AB - Cerebrovascular disease is highly heterogeneous but can culminate in vascular cognitive impairment or vascular dementia (VaD). As much as the clinical diagnosis warrants scrutiny, the neuropathological substrates of VaD also need to be better defined. Atherosclerosis and small vessel disease are the main causes of brain infarction. Lacunar infarcts or multiple microinfarcts in the basal ganglia, thalamus, brainstem and white matter are associated with more than half of VaD cases consistent with subcortical ischaemic VaD. White matter changes including regions of incomplete infarction are usually widespread in VaD, but their contribution to impairment is not explicit. Other pathologies including hippocampal injury and Alzheimer type of lesions may also modify the course of dementia. Similar to other common dementias consensus criteria for VaD need unambiguous definition to impact on preventative and treatment strategies and are critical for selective recruitment to clinical trials. PMID- 15537526 TI - Vascular cognitive disorder: a new diagnostic category updating vascular cognitive impairment and vascular dementia. AB - Vascular cognitive impairment (VCI) was proposed as an umbrella term to include subjects affected with any degree of cognitive impairment resulting from cerebrovascular disease (CVD), ranging from mild cognitive impairment (MCI) to vascular dementia. VCI may or may not exclude the host of "focal" circumscribed impairments of specialized functions such as language (aphasia), intentional gesture (apraxia), or categorical recognition (agnosia), among others, that may result from a stroke. Therefore, there are no universally accepted diagnostic criteria for VCI. We conclude that this concept could be more useful if it were to be limited to cases of vascular MCI without dementia, by analogy with the concept of amnestic MCI, currently considered the earliest clinically diagnosable stage of Alzheimer disease (AD). In agreement with our view,the Canadian Study on Health and Aging successfully implemented a restricted definition of VCI, excluding cases of dementia (i.e., vascular cognitive impairment no dementia, VCI ND). The Canadian definition and diagnostic criteria could be utilized for future studies of VCI. This definition excludes isolated impairments of specialized cognitive functions. Vascular dementia (VaD): The main problem of this diagnostic category stems from the currently accepted definition of dementia that requires memory loss as the sine qua non for the diagnosis. This may result in over sampling of patients with AD worsened by stroke (AD+CVD). This problem was minimized in controlled clinical trials of VaD by excluding patients with a prior diagnosis of AD, those with pre-existing memory loss before the index stroke, and those with amnestic MCI. We propose a definition of dementia in VaD based on presence of abnormal executive control function, severe enough to interfere with social or occupational functioning. Vascular cognitive disorder (VCD): This term, proposed by Sachdev [P. Sachdev, Vascular cognitive disorder. Int J Geriat Psychiatry 14 (1999)402-403.] would become the global diagnostic category for cognitive impairment of vascular origin, ranging from VCI to VaD. It would include specific disease entities such as post-stroke VCI, post-stroke VaD, CADASIL, Binswanger disease, and AD plus CVD. This category explicitly excludes isolated cognitive dysfunctions such as those mentioned above. PMID- 15537532 TI - Reenthronement of the muscle satellite cell. AB - In this issue of Cell (Sherwood et al., 2004), a quantitative survey confirms that repair of skeletal muscle is overwhelmingly attributable to the endogenous satellite cell population but that experience of a regenerating muscle environment confers some myogenic qualities onto a tiny population of bone marrow derived cells. PMID- 15537533 TI - HDAC4: a corepressor controlling bone development. AB - RUNX2 is a transcription factor with a well-characterized role in bone development. In this issue of Cell, Vega and colleagues (Vega et al., 2004) show that HDAC4 interacts with RUNX2 and impacts upon chondrocyte hypertrophy and bone formation. PMID- 15537534 TI - Silence of the rings. AB - Two recent papers (de Napoles et al., 2004 and Wang et al., 2004) have linked monoubiquitylation of histone H2A to the activities of E3 ubiquitin ligases that reside in Polycomb-group repressor complexes. PMID- 15537535 TI - Heterotrimeric G proteins: new tricks for an old dog. AB - Heterotrimeric G proteins are well known for their function in signal transduction downstream of seven transmembrane receptors. More recently, however, genetic analysis in C. elegans and in Drosophila has revealed a second, essential function of these molecules in positioning the mitotic spindle and attaching microtubules to the cell cortex. Five new publications in Cell (Afshar et al., 2004; Du and Macara, 2004 [this issue of Cell]; Hess et al., 2004), Developmental Cell (Martin-McCaffrey et al., 2004), and Current Biology (Couwenbergs et al., 2004) show that this function is conserved in vertebrates and--like the classical pathway--involves cycling of G proteins between GDP and GTP bound conformations. PMID- 15537536 TI - TROSPA, an Ixodes scapularis receptor for Borrelia burgdorferi. AB - The Lyme disease agent Borrelia burgdorferi naturally persists in a cycle that primarily involves ticks and mammals. We have now identified a tick receptor (TROSPA) that is required for spirochetal colonization of Ixodes scapularis. B. burgdorferi outer surface protein A, which is abundantly expressed on spirochetes within the arthropod and essential for pathogen adherence to the vector, specifically bound to TROSPA. TROSPA mRNA levels in ticks increased following spirochete infestation and decreased in response to engorgement, events that are temporally linked to B. burgdorferi entry into and egress from the vector. The blockade of TROSPA by TROSPA antisera or by the repression of TROSPA expression via RNA interference reduced B. burgdorferi adherence to the I. scapularis gut in vivo, thereby preventing efficient colonization of the vector and subsequently reducing pathogen transmission to the mammalian host. Identification of an I. scapularis receptor for B. burgdorferi is the first step toward elucidating arthropod ligands that are required for survival of spirochetes in nature. PMID- 15537537 TI - Heterochromatin regulates cell type-specific long-range chromatin interactions essential for directed recombination. AB - Mating-type switching in Schizosaccharomyces pombe involves replacing genetic information at the expressed mat1 locus with sequences copied from one of two silent donor loci, mat2-P or mat3-M, located within a 20-kb heterochromatic domain. Donor selection is dictated by cell type: mat2 is the preferred donor in M cells, and mat3 is the preferred donor in P cells. Here we show that a recombination-promoting complex (RPC) containing Swi2 and Swi5 proteins exhibits cell type-specific localization pattern at the silent mating-type region and this differential localization modulates donor preference during mating-type switching. In P cells, RPC localization is restricted to a recombination enhancer located adjacent to mat3, but in M cells, RPC spreads in cis across the entire silent mating-type interval in a heterochromatin-dependent manner. Our analyses implicate heterochromatin in long-range regulatory interactions and suggest that heterochromatin imposes at the mating-type region structural organization that is important for the donor-choice mechanism. PMID- 15537538 TI - Structural basis of transcription: nucleotide selection by rotation in the RNA polymerase II active center. AB - Binding of a ribonucleoside triphosphate to an RNA polymerase II transcribing complex, with base pairing to the template DNA, was revealed by X-ray crystallography. Binding of a mismatched nucleoside triphosphate was also detected, but in an adjacent site, inverted with respect to the correctly paired nucleotide. The results are consistent with a two-step mechanism of nucleotide selection, with initial binding to an entry (E) site beneath the active center in an inverted orientation, followed by rotation into the nucleotide addition (A) site for pairing with the template DNA. This mechanism is unrelated to that of single subunit RNA polymerases and so defines a new paradigm for the large, multisubunit enzymes. Additional findings from these studies include a third nucleotide binding site that may define the length of backtracked RNA; DNA double helix unwinding in advance of the polymerase active center; and extension of the diffraction limit of RNA polymerase II crystals to 2.3 A. PMID- 15537539 TI - She2p is a novel RNA binding protein with a basic helical hairpin motif. AB - Selective transport of mRNAs in ribonucleoprotein particles (mRNP) ensures asymmetric distribution of information within and among eukaryotic cells. Actin dependent transport of ASH1 mRNA in yeast represents one of the best characterized examples of mRNP translocation. Formation of the ASH1 mRNP requires recognition of zip code elements by the RNA binding protein She2p. We determined the X-ray structure of She2p at 1.95 A resolution. She2p is a member of a previously unknown class of nucleic acid binding proteins, composed of a single globular domain with a five alpha helix bundle that forms a symmetric homodimer. After demonstrating potent, dimer-dependent RNA binding in vitro, we mapped the RNA binding surface of She2p to a basic helical hairpin in vitro and in vivo and present a mechanism for mRNA-dependent initiation of ASH1 mRNP complex assembly. PMID- 15537540 TI - Mammalian Pins is a conformational switch that links NuMA to heterotrimeric G proteins. AB - During asymmetric cell divisions, mitotic spindles align along the axis of polarization. In invertebrates, spindle positioning requires Pins or related proteins and a G protein alpha subunit. A mammalian Pins, called LGN, binds Galphai and also interacts through an N-terminal domain with the microtubule binding protein NuMA. During mitosis, LGN recruits NuMA to the cell cortex, while cortical association of LGN itself requires the C-terminal Galpha binding domain. Using a FRET biosensor, we find that LGN behaves as a conformational switch: in its closed state, the N and C termini interact, but NuMA or Galphai can disrupt this association, allowing LGN to interact simultaneously with both proteins, resulting in their cortical localization. Overexpression of Galphai or YFP-LGN causes a pronounced oscillation of metaphase spindles, and NuMA binding to LGN is required for these spindle movements. We propose that a related switch mechanism might operate in asymmetric cell divisions in the fly and nematode. PMID- 15537541 TI - Structure of the Cand1-Cul1-Roc1 complex reveals regulatory mechanisms for the assembly of the multisubunit cullin-dependent ubiquitin ligases. AB - The SCF ubiquitin ligase complex regulates diverse cellular functions by ubiquitinating numerous protein substrates. Cand1, a 120 kDa HEAT repeat protein, forms a tight complex with the Cul1-Roc1 SCF catalytic core, inhibiting the assembly of the multisubunit E3 complex. The crystal structure of the Cand1-Cul1 Roc1 complex shows that Cand1 adopts a highly sinuous superhelical structure, clamping around the elongated SCF scaffold protein Cul1. At one end, a Cand1 beta hairpin protrusion partially occupies the adaptor binding site on Cul1, inhibiting its interactions with the Skp1 adaptor and the substrate-recruiting F box protein subunits. At the other end, two Cand1 HEAT repeats pack against a conserved Cul1 surface cleft and bury a Cul1 lysine residue, whose modification by the ubiquitin-like protein, Nedd8, is able to block Cand1-Cul1 association. Together with biochemical evidence, these structural results elucidate the mechanisms by which Cand1 and Nedd8 regulate the assembly-disassembly cycles of SCF and other cullin-dependent E3 complexes. PMID- 15537542 TI - Ferritin heavy chain upregulation by NF-kappaB inhibits TNFalpha-induced apoptosis by suppressing reactive oxygen species. AB - During inflammation, NF-kappaB transcription factors antagonize apoptosis induced by tumor necrosis factor (TNF)alpha. This antiapoptotic activity of NF-kappaB involves suppressing the accumulation of reactive oxygen species (ROS) and controlling the activation of the c-Jun N-terminal kinase (JNK) cascade. However, the mechanism(s) by which NF-kappaB inhibits ROS accumulation is unclear. We identify ferritin heavy chain (FHC)--the primary iron storage factor--as an essential mediator of the antioxidant and protective activities of NF-kappaB. FHC is induced downstream of NF-kappaB and is required to prevent sustained JNK activation and, thereby, apoptosis triggered by TNFalpha. FHC-mediated inhibition of JNK signaling depends on suppressing ROS accumulation and is achieved through iron sequestration. These findings establish a basis for the NF-kappaB-mediated control of ROS induction and identify a mechanism by which NF-kappaB suppresses proapoptotic JNK signaling. Our results suggest modulation of FHC or, more broadly, of iron metabolism as a potential approach for anti-inflammatory therapy. PMID- 15537543 TI - Isolation of adult mouse myogenic progenitors: functional heterogeneity of cells within and engrafting skeletal muscle. AB - Skeletal muscle regeneration in adults is thought to occur through the action of myogenic satellite cells located in close association with mature muscle fibers; however, these precursor cells have not been prospectively isolated, and recent studies have suggested that additional muscle progenitors, including cells of bone marrow or hematopoietic origin, may exist. To clarify the origin(s) of adult myogenic cells, we used phenotypic, morphological, and functional criteria to identify and prospectively isolate a subset of myofiber-associated cells capable at the single cell level of generating myogenic colonies at high frequency. Importantly, although muscle-engrafted cells from marrow and/or circulation localized to the same anatomic compartment as myogenic satellite cells and expressed some though not all satellite cell markers, they displayed no intrinsic myogenicity. Together, these studies describe the clonal isolation of functional adult myogenic progenitors and demonstrate that these cells do not arise from hematopoietic or other bone marrow or circulating precursors. PMID- 15537544 TI - Histone deacetylase 4 controls chondrocyte hypertrophy during skeletogenesis. AB - Histone deacetylases (HDACs) modulate cell growth and differentiation by governing chromatin structure and repressing the activity of specific transcription factors. We showed previously that HDAC9 acts as a negative regulator of cardiomyocyte hypertrophy and skeletal muscle differentiation. Here we report that HDAC4, which is expressed in prehypertrophic chondrocytes, regulates chondrocyte hypertrophy and endochondral bone formation by interacting with and inhibiting the activity of Runx2, a transcription factor necessary for chondrocyte hypertrophy. HDAC4-null mice display premature ossification of developing bones due to ectopic and early onset chondrocyte hypertrophy, mimicking the phenotype that results from constitutive Runx2 expression in chondrocytes. Conversely, overexpression of HDAC4 in proliferating chondrocytes in vivo inhibits chondrocyte hypertrophy and differentiation, mimicking a Runx2 loss-of-function phenotype. These results establish HDAC4 as a central regulator of chondrocyte hypertrophy and skeletogenesis and suggest general roles for class II HDACs in the control of cellular hypertrophy. PMID- 15537545 TI - Magnitude of binocular vision controlled by islet-2 repression of a genetic program that specifies laterality of retinal axon pathfinding. AB - Pathfinding of retinal ganglion cell (RGC) axons at the midline optic chiasm determines whether RGCs project to ipsilateral or contralateral brain visual centers, critical for binocular vision. Using Isl2tau-lacZ knockin mice, we show that the LIM-homeodomain transcription factor Isl2 marks only contralaterally projecting RGCs. The transcription factor Zic2 and guidance receptor EphB1, required by RGCs to project ipsilaterally, colocalize in RGCs distinct from Isl2 RGCs in the ventral-temporal crescent (VTC), the source of ipsilateral projections. Isl2 knockout mice have an increased ipsilateral projection originating from significantly more RGCs limited to the VTC. Isl2 knockouts also have increased Zic2 and EphB1 expression and significantly more Zic2 RGCs in the VTC. We conclude that Isl2 specifies RGC laterality by repressing an ipsilateral pathfinding program unique to VTC RGCs and involving Zic2 and EphB1. This genetic hierarchy controls binocular vision by regulating the magnitude and source of ipsilateral projections and reveals unique retinal domains. PMID- 15537546 TI - Developing local public health capacity in cultural competency: a case study with Haitians in a rural community. AB - With the increase in cultural diversity in all regions of the United States, public health employees and other health care providers are more challenged than ever before to provide culturally sensitive care to citizens from an increasing range of cultures. Building on their existing experience and skill, health care staff must expand their competencies to address this changing diversity in their communities. This project serves as a case study of how 1 small rural health department in Delaware created a Web site using Purnell's model for cultural competence (Purnell & Paulanka, 2003) as the organizational framework and rose to the challenge of assisting public health staff to provide culturally sensitive health services to a rapidly increasing population of Haitian immigrants. PMID- 15537547 TI - Cultural considerations in understanding family violence among Asian American Pacific Islander families. AB - There has been much research conducted in the area of family violence, however, there is a paucity of research specifically addressing family violence in Asian American and Pacific Islander (AAPI) communities. Because AAPIs are regarded as the model minority, many have the misconception that family violence does not exist in this population. This article examines Asian cultural beliefs and values to gain a better understanding of Asian attitudes towards family violence. A more in-depth exploration of the Cambodian and Vietnamese cultures regarding attitudes towards violence is included because of the higher tolerance of family violence among these 2 cultures than other Asian cultures. Public health nurses working with AAPI families can provide culturally appropriate care by taking Asian attitudes, values, and beliefs into account when developing nursing interventions with victims of abuse. PMID- 15537548 TI - Evaluating the effects of an educational symposium on knowledge, impact, and self management of older african americans living with osteoarthritis. AB - The purpose of the study1 was to determine the effects of a 1-day educational symposium on knowledge, impact, and self-management of older African Americans living with osteoarthritis (OA). A 1-group pre- and posttest design was employed. The educational symposium was the intervention variable. Speakers were experts in the field of arthritis care. Participants were recruited from a community senior center. Participants completed the Short Form of the Arthritis Impact Management Scales (AIMS2-SF), the Summary of Arthritis Management Methods (SAMMS), and a 6 question Knowledge Test about OA before the symposium. The knowledge test was administered again at the completion of the program. Posttesting of the AIMS2-SF and SAMMS occurred 3 months later to assess changes in arthritis impact and self management strategies. Knowledge scores increased significantly immediately following the symposium. The AIMS2-SF subscales of Affect, Symptom, and Physical Function decreased significantly 3 months after the symposium indicating that arthritis had less impact on participants after the conference. Participants also reported using more self-management methods 3 months after the event. The results of this pilot study indicate that this symposium was effective in increasing participants' knowledge of OA, improving self-management, and decreasing the impact of OA on daily function. The small sample size and preexperimental design limit generalizability. PMID- 15537549 TI - Pregnant adolescent reflections of parental communication. AB - The purpose of this study is to describe reflections of parental communication from the perspective of pregnant adolescents prior to pregnancy and during pregnancy. This qualitative design, using grounded theory methodology and semistructured interviews collected data from a convenience sample of predominantly Hispanic adolescent women (n = 30) in 4 settings. Participants were recruited from 3 area clinics and a transitional high school in California. The findings identified 7 themes from the interview data to support the adolescent perceptions of poor communication and unstable relationships with parents prior to the pregnancy and difficulty in communicating about the pregnancy to parents. Most participants identified a significant improvement in relationships and communication when pregnancy occurred. It is important for nurses to incorporate family communication strategies in their adolescent health interventions to help decrease the adolescent pregnancy rate. PMID- 15537552 TI - Clinical pharmacy, pharmaceutical care, and the quality of drug therapy. AB - BACKGROUND: Because of concerns about patient safety and the quality of health care in America, in particular about drug therapy, pharmacists have unprecedented opportunities to increase their value and significance. When defining clinical pharmacy and pharmaceutical care, pharmacists long ago recognized the need to improve the safety and effectiveness of drug therapy. OBJECTIVE: To describe how clinical pharmacy and pharmaceutical care, closely related concepts, can contribute to a strategy for improving the quality of drug therapy. DESIGN: Commentary and review of selected publications. CONCLUSION: Pharmacists can improve the quality of drug therapy by improving the organizational structures through which drug therapy is provided, specifically by creating medications use systems and by regularly evaluating their performance. As envisaged by the Institute of Medicine, these systems must be patient centered, cooperative, and interprofessional. To maximize pharmacists' participation in such systems, pharmaceutical education should include courses in medications use systems as necessary counterparts to courses in pharmacotherapeutics. Clinical functions must be organized around patient need and directed at outcomes. Clinical practice should constitute the mainstream practice of pharmacy rather than an "optional" specialty. Pharmaceutical care describes the original purpose of clinical pharmacy, when it was understood as a professional practice rather than a health science. It describes a way that clinical pharmacy, especially specialists and subspecialists, could coordinate their work more effectively. The concept of clinical pharmacy adds essential clarity about the process component of pharmacists' participation in, and strengthens the academic basis of, pharmaceutical care. The clinical, humanitarian, and economic case for preventing drug-related morbidity is strong, and pharmacy has much to offer. It is, again, time to work together as a profession to plan our common future. PMID- 15537553 TI - Clinical pharmacy and pharmaceutical care. PMID- 15537554 TI - Bioavailability and lack of toxicity of S-adenosyl-L-methionine (SAMe) in humans. AB - STUDY OBJECTIVE: To determine if S-adenosyl-L-methionine (SAMe), a widely used dietary supplement with antidepressant properties, is significantly bioavailable, and whether toxic methylated compounds are produced with oral SAMe administration in humans. Serum homocysteine levels were also measured since alterations in these levels have been theorized in association with SAMe. DESIGN: Unblinded pharmacokinetic trial. SUBJECTS: Fifteen healthy volunteers. SETTING: Clinical research unit in a psychiatric hospital. INTERVENTION: Subjects received oral SAMe for 4 weeks; the dosage was titrated over 5 days to 1600 mg/day. Serum levels of SAMe, toxic methylated compounds (methanol, formaldehyde, and formic acid), and homocysteine were measured at baseline and at weeks 2 and 4. At baseline, a structured clinical interview for axis I disorders (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition) was completed to assess for any undiagnosed psychiatric disorders. Mood was rated at baseline and at weeks 2 and 4 using the Zung Depression Rating Scale, Young Mania Rating Scale, Montgomery-Asberg Depression Rating Scale, Clinical Global Impression Scale, and the Global Assessment of Function Scale. MEASUREMENTS AND MAIN RESULTS: After oral administration, SAMe levels were significantly elevated. Slight, likely insignificant, elevations in serum formaldehyde levels were detected in three subjects. No subject exhibited elevated homocysteine levels during SAMe treatment. One subject developed a transient mixed manic state with suicidal ideation within 2 weeks of starting SAMe; she recovered fully within 3 days of discontinuing the compound. CONCLUSION: Oral dosages of 1600 mg/day of SAMe appear to be significantly bioavailable and nontoxic, at least regarding toxic methylated metabolites and homocysteine. However, the risk of mania in vulnerable individuals remains a serious concern. PMID- 15537555 TI - The influence of St. John's wort on the pharmacokinetics and protein binding of imatinib mesylate. AB - STUDY OBJECTIVE: To determine the effect of St. John's wort on the pharmacokinetics of imatinib mesylate. DESIGN: Open-label, complete crossover, fixed-sequence, pharmacokinetic study. SETTING: Clinical research center. SUBJECTS: Ten healthy adult volunteers. INTERVENTION: Single 400-mg oral doses of imatinib were administered before and after 2 weeks of treatment with St. John's wort 300 mg 3 times/day. MEASUREMENTS AND MAIN RESULTS: The pharmacokinetics of imatinib were significantly altered by St. John's wort, with reductions of 32% in the median area under the concentration-time curve from time zero to infinity (p=0.0001), 29% in maximum observed concentration (p=0.005), and 21% in half-life (p=0.0001). Protein binding ranged from 97.7-90.3% (mean 94.9%), was concentration independent, and was not altered by St. John's wort. Therapeutic outcomes of imatinib have been shown to correlate with both dose and drug concentrations. CONCLUSION: Coadministration of imatinib with St. John's wort may compromise imatinib's clinical efficacy. PMID- 15537556 TI - Work-related outcomes after a myocardial infarction. AB - STUDY OBJECTIVE: To evaluate work-related outcomes of patients at 7 months after a myocardial infarction and to identify patient, disease, and intervention characteristics associated with these outcomes. DESIGN: Cross-sectional survey analysis. SETTING: Large Midwestern academic health system. PATIENTS: Eighty-nine patients with the discharge diagnosis of acute myocardial infarction during a 1 year index period. INTERVENTION: Work performance questionnaire administered by telephone, and medical record review. MEASUREMENTS AND MAIN RESULTS: Seven months after discharge, 232 patients were interviewed by telephone to determine work status before and after myocardial infarction, work-related outcomes (absenteeism and perceived work performance, assessed by the Work Performance Scale [WPS] of the Functional Status Questionnaire), and health-related quality of life. Univariate analyses were used to determine the association between individual characteristics and work-related outcomes. Of the 89 patients who had worked before the index myocardial infarction, 21 (23.6%) did not return to work. Variables associated with the outcome of not returning to work were past myocardial infarction (before the index myocardial infarction), coronary artery bypass graft surgery, heart failure, positive stress test, and low score on the Physical Component Summary (PCS-12) scale of the Short Form-12. Patients who did not return to work also tended to have more comorbidities and take more prescribed drugs than those who returned to work. Median WPS scores were higher for patients who had higher ejection fractions at discharge, had not experienced a myocardial infarction before the index event, underwent a percutaneous revascularization intervention at the time of hospitalization, and had not recently been absent from work. Workers reporting absences had lower PCS-12 scores than their counterparts or reported a rehospitalization before the survey. CONCLUSION: Preexisting cardiac disease and poorer physical functioning were consistently related to worse work-related outcomes. This small study demonstrates the need for a larger, broader study that includes health beliefs, treatment, and other job and patient factors that may influence work-related outcomes. PMID- 15537558 TI - Frequency of new-onset diabetes mellitus and use of antipsychotic drugs among Central Texas veterans. AB - STUDY OBJECTIVES: To determine whether the frequency of new-onset diabetes mellitus differs between patients taking atypical antipsychotic agents and those taking typical agents, whether the frequency of new-onset diabetes differs among those taking the atypical antipsychotic agents, and what clinical and demographic factors influence the occurrence of new-onset diabetes. DESIGN: Retrospective analysis. SETTING: Central Texas Veterans Health Care System. PATIENTS: Continuously enrolled adult (> or = 18 yrs) patients with no previous (6 mo) antipsychotic use and no history (previous 1 yr) of diabetes. MEASUREMENTS AND MAIN RESULTS: Data from the Central Texas Veterans Health Care System were extracted from September 1995-November 2002. Clinical and demographic factors used in the analysis were antipsychotic agent taken, body mass index, diabetes related risk factors, type of mental health comorbidity, age, sex, and race. Among those who met the inclusion criteria (3469 patients), chi2 analyses revealed no significant difference in the frequency of diabetes between the typical and atypical groups (p=0.5553) or among those taking atypical agents (p=0.6520). Multivariate logistic regression (1587 patients) revealed that increasing age (odds ratio [OR] 1.213, 95% confidence interval [CI] 1.016-1.447, p=0.0324), nonwhite race (OR 1.761, 95% CI 1.174-2.640, p=0.0062), and hyperlipidemia (OR 1.606, 95% CI 1.064-2.425, p=0.0242) were significantly related to new-onset diabetes. CONCLUSIONS: Among veterans taking antipsychotic agents, no difference was noted in the frequency of diabetes between patients who took typical agents and those who took atypical agents. After controlling for demographic and clinical variables, still no significant difference was noted among the agents. The main factors (increasing age, nonwhite race, and hyperlipidemia) related to new-onset diabetes were those that are typically associated with the disease. PMID- 15537557 TI - Bupropion may support psychosocial treatment of nicotine-dependent adolescents: preliminary results. AB - STUDY OBJECTIVE: To assess the efficacy and safety of long-term bupropion therapy for nicotine dependence in adolescents. DESIGN: Prospective, double-blind, placebo-controlled trial. SETTING: Outpatient clinic in Innsbruck, Austria. SUBJECTS: Twenty-two adolescents, aged 16-19 years, with nicotine dependence. INTERVENTION: Participants were randomly assigned to receive bupropion 150 mg/day or placebo for 90 days. MEASUREMENTS AND MAIN RESULTS: Patients were classified as abstinent or relapsed on day 0 (when study drug was started) and again on days 30 and 90, according to their self-reports. Treatment failure was defined as relapse or nonattendance. Time to first treatment failure was the primary outcome measure. Mean cumulative abstinence duration was significantly greater in the bupropion group than in the placebo group (78.4 +/- 39.6 vs 30.2 +/- 19.2 days. p=0.0042). CONCLUSION: Bupropion seems to be an effective and well-tolerated pharmacologic adjunct to psychosocial and behavioral treatment programs for some adolescent nicotine-dependent patients. However, experienced clinicians should continuously monitor patients for adverse effects. PMID- 15537559 TI - Magnitude and duration of elevated gastric pH in patients infected with human immunodeficiency virus after administration of chewable, dispersible, buffered didanosine tablets. AB - STUDY OBJECTIVES: To test the hypothesis that gastric pH would be elevated above pH 3.0 for at least 2 hours after administration of chewable, dispersible, buffered didanosine tablets. Doses tested were 200 mg (two 100-mg tablets) and 400 mg (two 200-mg tablets). We also sought to compare these doses with regard to maximum gastric pH (pHmax), time to pHmax (TpH-max), time that gastric pH exceeds 3.0 (TpH>3), and area under the gastric pH versus time curve for pH greater than 3.0 (AUCT>pH 3). DESIGN: Prospective, parallel-group, dose-comparison, gastric pH study. SETTING: General Clinical Research Center, University of Michigan Hospitals, Ann Arbor, Michigan. PATIENTS: Nineteen patients infected with human immunodeficiency virus, aged 30-62 years, and receiving long-term didanosine therapy. INTERVENTION: Patients underwent continuous gastric pH monitoring, using the Heidelberg capsule radiotelemetric pH monitoring device. After documentation of a fasting baseline gastric pH below 3.0, patients were given 180 ml of water (control phase), and gastric pH was allowed to return to baseline. After administration of a single, oral dose of didanosine 200 mg or 400 mg with 180 ml of water, gastric pH was recorded until pH remained below 3.0 for 10 minutes. MEASUREMENTS AND MAIN RESULTS: A mean pHmax of 8.6 (range 6.3-9.5) was achieved with a TpH-max of 4.1 minutes (range 1-12.0 min). Mean TpH>3 was 24.9 minutes (range 15-55 min), with an AUCT>pH 3 of 2.6 pH x min(-1) (range 1.2-6.9 pH x min( 1)). The two doses of didanosine tested did not differ significantly in mean gastric pH parameters. CONCLUSIONS: After administration of chewable, dispersible, buffered didanosine tablets, 200 or 400 mg, the mean duration of elevated gastric pH (TpH>3) was less than 30 minutes, with a range of 15-55 minutes. Characterization of the magnitude and duration of elevated gastric pH may allow for earlier administration of other pH-sensitive drugs. The short duration of elevated gastric pH may help explain the wide variability in didanosine bioavailability observed clinically. PMID- 15537560 TI - Follow-up of adverse drug reactions from peginterferon alfa-2b-ribavirin therapy. AB - STUDY OBJECTIVE: To investigate the adverse drug reactions (ADRs) from peginterferon alfa-2b plus ribavirin in patients infected with the hepatitis C virus (HCV). DESIGN: Prospective, observational, pharmacovigilance study. SETTING: Gastroenterology department of a French university hospital. PATIENTS: A cohort of consecutive outpatients who were treated with peginterferon alfa-2b plus ribavirin for viral hepatitis. INTERVENTION: During the 1-year period of HCV therapy visits, a medical staff member trained in pharmacovigilance interviewed the patients about all ADRs and their use of other drugs. The ADRs assessed as serious were confirmed from regular review of the medical records. MEASUREMENTS AND MAIN RESULTS: A total of 87 patients were treated for HCV. Among these, peginterferon alfa-2b plus ribavirin therapy was started in 51 patients; one patient was lost to follow-up after 1 month. The ADRs were assessed as serious in 10 patients (20%): suicide (one patient), hospitalization (three patients), and definitive discontinuation of peginterferon alfa-2b plus ribavirin (six patients). The ADRs contributed to or were the main reason for withdrawing HCV drugs in 8 patients (16%). Dosage reductions of ribavirin and/or peginterferon alfa-2b were required in 10 patients (20%) and seemed less frequent than that needed in clinical trials. Compared with results of clinical trials, a similar frequency of hair loss, higher frequency of injection-site reactions, lower frequency of depression or insomnia, and higher frequency of endocrine ADRs or blurred vision were detected. CONCLUSION: Results suggest some differences in the frequencies of ADRs compared with data from clinical trials. A longer period of monitoring is warranted to improve knowledge about ADRs of pegylated interferon. A medical staff member trained in pharmacovigilance is useful to collect quantitative and qualitative data about ADRs. PMID- 15537561 TI - Drug-eluting stents: a mechanical and pharmacologic approach to coronary artery disease. AB - Coronary artery disease is the largest killer of men and women in the United States and costs the health care system billions of dollars annually. Several advances in both mechanical and pharmacologic treatment of coronary artery disease have occurred in recent decades. Mechanically, percutaneous coronary intervention is commonly used to treat coronary atherosclerosis. This approach has dramatically reduced both morbidity and mortality for patients with different levels of severity of coronary artery disease. However, percutaneous coronary intervention is limited by restenosis, which is an increase in growth of the intimal layer of the vessel wall. Despite the introduction of intracoronary stents and the addition of systemic pharmacotherapy, restenosis still affects a significant number of patients. The new technology of drug-eluting stents combines mechanical and pharmacologic approaches to prevent restenosis. Various types of these stents exist in different stages of development; several have been shown to prevent or reduce intimal growth after stent deployment. An understanding of how this combined mechanical and pharmacologic approach reduces restenosis requires consideration of complex issues in pathophysiology and pharmacology. PMID- 15537562 TI - Intravenous ethanol for the treatment of alcohol withdrawal syndrome in critically ill patients. AB - Critically ill patients with alcoholism are at greater risk of morbidity and mortality from alcohol withdrawal syndrome than are patients without alcoholism. Benzodiazepines are considered the drugs of choice for the prevention and treatment of alcohol withdrawal syndrome, but some studies have suggested that intravenous ethanol may be as effective as those agents, as well as being less sedating. We evaluated the evidence regarding the use of intravenous ethanol for the prevention and treatment of alcohol withdrawal syndrome in critically ill patients in order to determine its role in this patient population. Because of the paucity of well-designed clinical trials, and because of intravenous ethanol's questionable efficacy, inconsistent pharmacokinetic profile, and relatively narrow therapeutic index, routine use of this drug is not recommended in critically ill patients who have alcohol withdrawal syndrome or are at risk for it. PMID- 15537563 TI - Key references in infectious diseases pharmacotherapy. AB - Most health care practitioners are challenged to maintain knowledge of contemporary practice issues in many therapeutic disciplines. Like many other areas, infectious diseases pharmacotherapy continues to evolve because of new information regarding disease epidemiology and new treatment options. Emerging infections and resistance further compound the need for information. To assist clinicians in identifying such important new information, we compiled a list of key references on infectious diseases pharmacotherapy published over the last 2 years. PMID- 15537564 TI - High-impact articles related to the pharmacotherapeutic management of systolic heart failure. AB - This compilation is part of a series of five articles identifying important literature in cardiovascular pharmacotherapy. This list focuses on pharmacotherapeutic management of acute decompensated and chronic heart failure. Most of the cited works present the results of landmark clinical studies that have shaped the management of patients with left ventricular systolic dysfunction. Limited primary literature is available for some topics; thus, pertinent review articles also are listed. In addition, consensus documents formed by expert panels in the United States and Europe are reviewed. This compilation may serve as a teaching tool, reference resource, or update of the literature for pharmacy clinicians, physicians, and students. PMID- 15537565 TI - Interaction between nesiritide and a heparin-coated pulmonary artery line during an infusion for decompensated heart failure. AB - STUDY OBJECTIVE: Because it is known that intravenous nesiritide is not compatible with unfractionated heparin, we sought to determine the effect that heparin coating on a pulmonary artery catheter may have on the efficacy of a nesiritide infusion. METHODS: The efficacy of a nesiritide infusion given through a heparin-coated pulmonary artery line was compared with that of a nesiritide infusion administered in the same patient through a heparin-free peripheral line. RESULTS: The rate of infusion was titrated to maintain consistent hemodynamic parameters. When nesiritide was administered through a heparin-coated line, the infusion rate escalated from 0.01 microg/kg/minute to 0.07 microg/kg/minute. After the route of administration was switched to a heparin-free line, the same hemodynamic parameters were maintained. The heparin-free line made it possible to reduce the infusion rate by 57.1% over the next 24 hours to 0.03 microg/kg/minute. CONCLUSION: The interaction of nesiritide with heparin-coated pulmonary artery lines has the potential to be clinically significant. Clinicians should be educated about this potential interaction. Nesiritide should be infused only through heparin-free lines. PMID- 15537566 TI - Early evaluation of hematuria in a patient receiving anticoagulant therapy and detection of malignancy. AB - A 63-year-old Caucasian man had a painless episode of dark-colored urine while taking warfarin 62.5 mg/week for lone atrial fibrillation in the presence of documented stable anticoagulation. Urinalysis revealed microscopic hematuria. Three weeks later, he had an episode of gross, painless hematuria. Thorough evaluation of the upper and lower urinary tract with renal ultrasound, intravenous pyelography, and cystoscopy revealed poorly differentiated, early stage, transitional cell carcinoma of the bladder. The patient was not aware of any exposure to carcinogens known to predispose to bladder cancer, nor was he a tobacco user. Early identification of the malignancy allowed for aggressive surgical intervention. Although this patient was considered low risk for the development of bladder cancer and was taking anticoagulants, the presence of hematuria was indicative of underlying pathology. Timely and thorough evaluation of hematuria in patients taking anticoagulants is necessary to identify and treat clinically important pathology. PMID- 15537567 TI - Empiric treatment of multidrug-resistant Burkholderia cepacia lung exacerbation in a patient with cystic fibrosis: application of pharmacodynamic concepts to meropenem therapy. AB - A 31-year-old man with cystic fibrosis was diagnosed with multidrug-resistant Burkholderia cepacia pneumonia. Meropenem 2000 mg every 8 hours was administered as a 3-hour infusion to maximize pharmacodynamic exposure; oral minocycline 100 mg twice/day was also given. Blood samples were collected to confirm meropenem concentrations. Concentrations above the mimimum inhibitory concentration (MIC) of 8 microg/ml were achieved for 52% of the dosing interval, which is greater than what is required for a bactericidal effect. The patient's condition improved, he was discharged, and completed a 3-week course of the antibiotic regimen. After 6 months, he had remained at his baseline level of health. This case demonstrates that pharmacodynamic principles can be used to design an antibiotic dosing regimen that can achieve optimal exposures when the MIC is above that considered susceptible to conventional dosing strategies. PMID- 15537568 TI - Benefit of ribose in a patient with fibromyalgia. AB - Ribose was added to the existing treatment regimen of a woman with fibromyalgia, resulting in a decrease in symptoms. It has been postulated that patients with fibromyalgia may have an alteration in muscle adenine nucleotide metabolism, leading to depleted energy reserves and an imbalance in cellular adenosine 5' triphosphate:adenosine 5'-diphosphate:adenosine 5'-monophosphate (ATP:ADP:AMP) ratios with an abnormal energy charge. As a key component in adenine nucleotide synthesis, ribose supplementation may be useful in such patients. PMID- 15537569 TI - Tegaserod-induced myocardial infarction: case report and hypothesis--an alternative viewpoint. PMID- 15537570 TI - The use of antibiotics as secondary prevention for cardiac events. PMID- 15537571 TI - Activation of peroxisome proliferator-activated receptor alpha increases the expression and activity of microsomal triglyceride transfer protein in the liver. AB - Microsomal triglyceride transfer protein (MTP) is rate-limiting in the assembly and secretion of lipoproteins containing apolipoprotein (apo) B. Previously we demonstrated that Wy 14,643 (Wy), a peroxisome proliferator-activated receptor (PPAR) alpha agonist, increases apoB-100 secretion despite decreased triglyceride synthesis. In this study, we sought to determine whether PPARalpha activation increases MTP expression and activity. Treatment with Wy increased hepatic MTP expression and activity in rats and mice and increased MTP expression in primary cultures of rat and mouse hepatocytes. Addition of actinomycin D blocked this increase and the MTP promoter (-136 to +67) containing a conserved DR1 element was activated by Wy, showing that PPARalpha activates transcription of the gene. Wy did not affect MTP expression in the intestine or in cultured hepatocytes from PPARalpha-null mice. A retinoid X receptor agonist (9-cis-retinoic acid), but not a PPARgamma agonist (rosiglitazone), increased MTP mRNA expression in cultured hepatocytes from both wild type and PPARalpha-null mice. In rat hepatocytes incubated with Wy, MTP mRNA levels increased between 6 and 24 h, and MTP protein expression and apoB-100 secretion increased between 24 and 72 h. In conclusion, PPARalpha activation stimulates hepatic MTP expression via increased transcription of the Mtp gene. This effect is paralleled by a change in apoB-100 secretion, indicating that the effect of Wy on apoB-100 secretion is mediated by increased expression of MTP. PMID- 15537572 TI - Mitochondrial release of pro-apoptotic proteins: electrostatic interactions can hold cytochrome c but not Smac/DIABLO to mitochondrial membranes. AB - A key step in the initiation of apoptosis is the release from the mitochondrial intermembrane space of cytochrome c and other pro-apoptotic proteins such as Smac/DIABLO, Omi/HtrA2, apoptosis-inducing factor (AIF), and endonuclease G (EndoG). Discrepancies have arisen, however, as to whether all these proteins are released in different systems. Our results suggest that failure to observe cytochrome c release may be due to the use of different buffers because after permeabilization by caspase-8 cleaved human Bid (tBid), cytochrome c dissociation from mitochondria was highly dependent on ionic strength and required 50-80 mm KCl, NaCl, or LiCl. In addition, mitochondria isolated from apoptotic cells using low ionic strength buffer bound a greater proportion of endogenous cytochrome c. In contrast to cytochrome c, Smac/DIABLO and Omi/HtrA2 were released independent of ionic strength, and AIF and EndoG behaved as if they are exposed to the intermembrane space but tethered to or within the inner membrane. AIF and EndoG were also not released by active caspases, which suggests their involvement in apoptosis may be limited. In summary, whereas tBid permeabilizes the outer membrane to cytochrome c, Smac/DIABLO, and Omi/HtrA2, the release of cytochrome c during apoptosis will be underestimated unless sufficient ionic strength is maintained to overcome the electrostatic association of cytochrome c with membranes. PMID- 15537573 TI - The "Welcome to Medicare" physical: a great opportunity for our seniors. PMID- 15537574 TI - The staging of colorectal cancer: 2004 and beyond. AB - Stage is the strongest predictor of survival for patients with colorectal cancer. Accurate staging also is critical for appropriate patient management and meaningful clinical research. Uniform staging criteria applied in a uniform manner are essential for accurate evaluation of therapies and outcomes. Historically, numerous different staging systems for colorectal cancer have been employed, but a single internationally recognized system is required to ensure a common language for cancer that is understood by clinicians in all specialties. For the tumor, node, metastasis system to remain relevant, it has to continuously undergo critical evaluation and change when clinically indicated. PMID- 15537575 TI - Identifying and testing for hereditary susceptibility to common cancers. AB - Hereditary cancer syndromes account for an estimated 5% of breast, ovarian, and colon cancers. The rapid discovery of cancer-related genes in the last 15 years has propelled the field of cancer genetic risk assessment forward. With patients becoming increasingly aware of available genetic testing options, it is important that various health professionals become knowledgeable in identifying and advising patients at increased risk for a hereditary cancer syndrome. This article will outline the components of providing a hereditary cancer risk assessment with a focus on hereditary breast and ovarian cancer syndrome and hereditary colon cancer. PMID- 15537577 TI - Performance and reporting of clinical breast examination: a review of the literature. AB - Clinical breast examination (CBE) seeks to detect breast abnormalities or evaluate patient reports of symptoms to find palpable breast cancers at an earlier stage of progression, when treatment is more effective and treatment options are greater than for later stage disease. Evidence suggests that, for some women, CBE can be an important complement to mammography in the earlier detection of breast cancer; CBE identifies some cancers missed by mammography and provides an important screening tool among women for whom mammography is not recommended or women who do not receive high-quality screening mammography according to recommended guidelines. But CBE performance and reporting approaches are inconsistent. Health care providers indicate that they are not confident in their CBE skills and would welcome training. Studies demonstrate that training can enhance CBE performance, measured in terms of execution of CBE components and accuracy. This literature review provides evidence to the extent that it is available, to support the specific recommendations of Saslow, et al. for optimizing CBE performance and reporting and to guide further research on CBE performance characteristics, reporting systems, barriers to high-quality CBE performance, and training. PMID- 15537576 TI - Clinical breast examination: practical recommendations for optimizing performance and reporting. AB - Clinical breast examination (CBE) seeks to detect breast abnormalities or evaluate patient reports of symptoms to find palpable breast cancers at an earlier stage of progression. Treatment options for earlier-stage cancers are generally more numerous, include less toxic alternatives, and are usually more effective than treatments for later-stage cancers. For average-risk women aged 40 and younger, earlier detection of palpable tumors identified by CBE can lead to earlier therapy. After age 40, when mammography is recommended, CBE is regarded as an adjunct to mammography. Recent debate, however, has questioned the contributions of CBE to the detection of breast cancer in asymptomatic women and particularly to improved survival and reduced mortality rates. Clinicians remain widely divided about the level of evidence supporting CBE and their confidence in the examination. Yet, CBE is practiced extensively in the United States and continues to be recommended by many leading health organizations. It is in this context that this report provides a brief review of evidence for CBE's role in the earlier detection of breast cancer, highlights current practice issues, and presents recommendations that, when implemented, could contribute to greater standardization of the practice and reporting of CBE. These recommendations may also lead to improved evidence of the nature and extent of CBE's contribution to the earlier detection of breast cancer. PMID- 15537578 TI - Patient pages. Colorectal cancer staging. PMID- 15537579 TI - Public perceptions of cost containment strategies: mixed signals for managed care. AB - With health care costs, and insurance premiums in particular, escalating rapidly, we may see the reintroduction of utilization management strategies associated with managed care, which seemed destined for oblivion only a short time ago. Results from a survey to assess Americans' views of managed care cost containment strategies indicate mixed support: Despite an overall lack of confidence in managed care, Americans appear to be receptive to specific managed care practices. Those designing cost containment strategies must find a balance between imposing restrictions that moderate use and hold down costs and allowing consumers to retain some control over their own health care. PMID- 15537580 TI - The challenges facing private health insurance. PMID- 15537581 TI - Monopoly, monopsony, and market definition: an antitrust perspective on market concentration among health insurers. AB - James Robinson uses the Herfindahl-Hirschman Index (HHI) to compute the concentration of commercial health insurance markets in most of the states during the past four years. The HHI is the analytical foundation for the federal antitrust merger guidelines, so we consider his findings from an antitrust perspective. Market concentration provides an important benchmark for antitrust analysis, but it does not, standing alone, indicate the presence of problematic (anticompetitive) behavior or a problem that antitrust law can solve. Even if it did, there are major problems in treating individual states as discrete insurance markets. Unless the market is correctly defined, any analysis of market concentration is thoroughly unreliable. PMID- 15537582 TI - Is there evidence that recent consolidation in the health insurance industry has adversely affected premiums? AB - James Robinson suggests that recent consolidation in the insurance market has been a cause of higher health insurance prices (premiums). Although the recent consolidation among health insurers and rising premiums are indisputable, it is unlikely that consolidation has had any adverse effect on premiums nationwide, and Robinson provides no data that suggest otherwise. Specifically, he does not present data showing an increase in concentration in any relevant market during the past few years, let alone any resulting increase in premiums. Health insurance consolidation in certain local markets could adversely affect premiums, but it seems clear that it is not a major national antitrust issue. PMID- 15537583 TI - Hot potato endgame. AB - Other stakeholders and events will influence whether health insurers' current postmerger prosperity will lead to U.S. health benefit programs that are predominantly sponsored by the private or public sectors. Large employers are encouraging three complementary health benefit innovations to improve the affordability and quality, or "efficiency," of clinical services: portable spending accounts, provider pay-for-performance, and tiered plans. If health insurers prefer private-sector health benefit sponsorship, they will need to implement these innovations robustly, despite the risks they pose to insurers' current but predictably temporary prosperity. Clinical efficiency gains can also sustain access to biomedical innovations for low- and moderate-income Americans. PMID- 15537584 TI - Professionalism reconsidered: physician payment in a small-practice environment. AB - Traditional fee-for-service health insurance rested on the assumption that doctors have primary responsibility for decisions about care. Managed competition assumed a new model based on corporate medicine, which has not materialized; also, consumers' ability to replace doctors as primary medical decisionmakers is unproven. Data on practice size show that doctors and patients continue to prefer the small-practice setting, where the doctor's role as the patient's agent is salient. The persistence of the small practice suggests that medical professionalism remains the cornerstone of the health system. If so, it may be more appropriate to pursue quality-oriented refinements of traditional payment approaches, rather than radical transformation. PMID- 15537585 TI - Professionalism reconsidered: physician payment from a health plan perspective. AB - Traditional fee-for-service health insurance rested on the assumption that doctors have primary responsibility for decisions about care. Those who long for unrestrained autonomy in an environment that is characterized by "information overload" and that continually demonstrates lapses in safety and quality fail to grasp the importance of physician and patient decision support systems and the key role that a health plan may play in dealing with these issues. The unbalanced allocation of resources between and among physician specialties is a contributing factor that must be addressed. PMID- 15537586 TI - Physician organization and care management in California: from cottage to Kaiser. AB - Data from a survey of practicing physicians in California's thirteen largest urban counties were used to ascertain differences in care management processes, financial incentives for quality, and practice pressures by type of practice setting. Physicians in the Permanente medical groups have adopted and value quality-oriented, system-level care management tools to a much greater degree than physicians in independent practice association (IPA) networks or traditional "cottage-industry" practices. Our findings raise disturbing questions about how the health system will close the "quality chasm" in medical care without transforming the underlying organization of physician practices. PMID- 15537587 TI - Does population management of chronic disease lead to lower costs of care? AB - The paper by Bruce Fireman and colleagues is an important contribution to the dialogue about the role of chronic disease management in quality improvement and health care cost mitigation. There is much enthusiasm for the potential impact of disease management techniques on the costs associated with chronic and complex health conditions. This Perspective describes several considerations that are important to the interpretation of studies of the cost impact of disease management and to assessments of the future usefulness of these interventions. PMID- 15537588 TI - The underwriting cycle: the rule of six. AB - The underwriting cycle is a thing of the past for most health insurance companies. There were six primary factors that caused the six-year pattern of the underwriting cycle for 1965-1991. These factors were claims payment cycle time, renewal dates and process, growth versus profit objectives, role of the actuary, rate regulation, and reimbursement methods. Most companies have made major changes to influence these factors, which will prevent a recurrence of the underwriting cycles of the past. PMID- 15537589 TI - Barriers to constraining health care cost growth. AB - Managing health care cost growth is a fundamental challenge facing our health care system. Through analysis of semistructured interviews, we conclude that barriers to health plan-level cost containment activities and strong forces outside the control of individual health plans will prevent many health system reforms (such as more competition among plans or modest increases in patient copayments) from stemming health care cost inflation. Policy debates and budgetary discussions must recognize that health care cost growth in excess of gross domestic product (GDP) growth is likely inevitable in the foreseeable future. The policy focus should be directed toward encouraging value. PMID- 15537590 TI - Doughnut-hole economics. AB - Both the recently enacted Medicare prescription drug benefit and a new cohort of consumer-directed health benefit models offer doughnut-shaped insurance coverage with large deductibles that begin around the mean annual spending for enrollees. These policies leave enrollees to bear more risk than policies with equal expected payouts that rely on first-dollar deductibles. This risk to enrollees is substantial, given the skewed distribution of health care spending and the placement of the typical deductible. I consider alternative explanations for this new benefit design trend and conclude that the desire to distribute tangible benefits to the largest number of constituents is most plausible. PMID- 15537591 TI - Stanford University's experience with managed competition. AB - Stanford University has a "managed competition" model of health insurance. Stanford contributes the cost of the low-cost plan, and employees are responsible for premium differences between this plan and other offerings. Each employee gets what he or she wants and is willing to pay for, and everyone has low-cost access to health insurance. Stanford risk-adjusts the premiums based on age and sex and plans soon to adjust including prescription drug data. In the past five years, premiums have risen rapidly, in line with the rest of the market. For competition to transform the delivery system, most employers in the region must adopt managed competition. PMID- 15537592 TI - Health plans' strategies to control prescription drug spending. AB - A number of recent studies have documented the sizable impact of consumer cost sharing without accounting for the other drug management strategies being adopted simultaneously. This qualitative case study of five of California's largest health plans examines the strategies and methods used to control prescription drug use and spending. Higher cost sharing is being used increasingly. Concurrently, major administrative efforts directed at physicians-including rules, incentives, and education-are being undertaken. These efforts have focused on lowering the cost per prescription by emphasizing generic substitution and therapeutic interchange of less costly drugs. PMID- 15537593 TI - Technology and the boundaries of the hospital: three emerging technologies. AB - U.S. hospitals have proved remarkably adept at altering their service offerings to incorporate new technologies. New technologies threatened to undercut hospitals' central role in health care delivery in the 1980s. An array of new technologies promise yet again to alter the boundaries of hospitals' franchise. These technologies will not only continue the shift away from acute, inpatient care that we have seen for the past thirty years but will also challenge hospitals to collaborate more effectively with physicians and technology developers. How hospitals and policymakers respond to these emerging technologies will help determine whether hospitals remain at the center of the U.S. health system. PMID- 15537594 TI - The decline in receipt of substance abuse treatment by the privately insured, 1992-2001. AB - This study examines underlying trends in substance abuse services financed by private insurance. Analyses are based on the 1992 and 2001 Medstat MarketScan database, a claims database from large employers. The percentage of beneficiaries using any substance abuse services declined by 23 percent from 1992 to 2001 (from 0.64 percent of enrollees to 0.49 percent of enrollees). This decline was evident in all categories: inpatient, outpatient, and pharmaceutical usage. Substance abuse spending per covered life (in constant dollars) dropped from about $21.16 in 1992 to about $5.58 in 2001 [corrected] PMID- 15537595 TI - Independent medical review of health plan coverage denials: early trends. AB - Concerns among patients and physicians that health plans would deny coverage for medically necessary services has brought about legislation mandating independent medical review (IMR) of denied services. Among 1,400 cases submitted in California, the most common areas for dispute were cancer care, endocrine/metabolic care (especially for obesity), orthopedic care, and neurological disorders. Surgery and pharmacy services constituted 52 percent of cases. In 58 percent of cases, IMR upheld the health plan's decision, while in 33 percent of cases the health plan's decision was overturned, which suggests that external IMR provides additional patient protection in California's health care system. PMID- 15537596 TI - Covering kids: variation in health insurance coverage trends by state, 1996-2002. AB - We estimated state-specific changes in health insurance coverage rates for children between 1996-1998 and 2001-2002. We found considerable variation in the changing distribution of health insurance coverage for children across states, with significant increases in public program coverage in twenty-nine states and significant decreases in uninsured children in twenty-seven. Children in families with incomes below 200 percent of the federal poverty level were the most likely to enroll in public programs. We provide an overview of state outreach and administrative simplification efforts and raise concerns about the persistent variation in children's health insurance coverage across states. PMID- 15537597 TI - Public expectations of nonprofit and for-profit ownership in American medicine: clarifications and implications. AB - Policymakers, advocates, and scholars frequently make claims about how the American public sees ownership affecting the delivery of medical care. In this paper we provide a comprehensive assessment of how Americans think about nonprofit and for-profit ownership. We summarize findings from surveys fielded between 1985 and 2000 and supplement them with findings from a new survey. Most Americans believe that ownership matters for multiple aspects of medical care; they expect nonprofit hospitals and health plans to be more trustworthy, fair, and humane but lower in quality. People who are better informed about ownership have more positive expectations about nonprofits' performance. PMID- 15537598 TI - Trends in contract management: the hidden evolution in hospital organization. AB - Contract management is an arrangement whereby the day-to-day operation of the hospital is contracted to an outside organization. In the past two decades there has been dramatic growth in the number of hospitals opting for contract management, yet surprisingly little attention has been paid to this phenomenon. Using national data, we report trends and demonstrate that adoption of contracts results in decreases in service offerings more often than increases. Since contract-managed hospitals tend to be located in rural areas, this raises concerns regarding access to care. On the other hand, contract management may allow distressed hospitals to survive. PMID- 15537599 TI - Acquainted with the night. PMID- 15537600 TI - A hospital death. PMID- 15537601 TI - Patterns of individual health insurance coverage, 1996-2000. AB - Information about patterns of individual health insurance coverage is limited. Knowledge gaps include the extent to which individual insurance provides transitional versus long-term coverage, and participants' insurance status before and after being covered by an individual plan. In this study we use data from the 1996-2000 Survey of Income and Program Participation (SIPP) to examine how long the individually insured maintain their coverage; sources of coverage before and after enrolling in an individual health plan; and characteristics of those who rely on individual insurance coverage. Understanding the dynamics of this market will better inform federal and state insurance reform efforts. PMID- 15537602 TI - Public perceptions of private health care in socialist China. AB - We present the findings of a United Nations Development Programme-World Health Organization study commissioned by China's Ministry of Health on use of public and private ambulatory care services in three Chinese provinces. We found much unmet medical need (16 percent), attributed mainly to the perceived high cost of care. Seventy-one percent had no health insurance (90 percent in rural and 51 percent in urban areas). For 33 percent, the last consultation was with a private practitioner. Widespread dissatisfaction with public providers (mainly high user fees and poor staff attitudes) is driving patients to seek cheaper but lower quality care from poorly regulated private providers. PMID- 15537603 TI - Quality oversight in medicaid primary care case management programs. AB - As health maintenance organizations (HMOs) have curtailed participation in Medicaid, enrollment in primary care case management (PCCM) programs has grown. To examine state Medicaid agencies' monitoring of PCCM and HMO programs, we surveyed Medicaid agency directors of forty-six states and the District of Columbia. Agencies were less likely to collect performance data in PCCM programs than in HMO programs. Few PCCM programs reported performance results for the public or providers. Reporting states tended to emphasize utilization results over quality-measure results. Despite growing enrollment, PCCM programs appear less likely to use the quality-oversight strategies employed by Medicaid health plans. PMID- 15537604 TI - Scarce physicians encounter scarce foundations: a call for action. AB - The United States is in the early phases of a deepening shortage of physicians, a situation last experienced fifty years ago. As then, energy and creativity will be needed to meet the nation's needs, and U.S. philanthropic foundations will again be called upon to play leadership roles. The issues are broad--extending from medical education to regulation and from building new schools to recruiting more international medical graduates. Throughout these issues, foundations are uniquely positioned to convene stakeholders, fund analyses, foster new medical education paradigms, and support the growth of its infrastructure. Foundations will be necessary partners in what is to come. PMID- 15537605 TI - Establishing, funding, and sustaining a university outreach program in oral health. AB - The Surgeon General's report of May 2000, Oral Health in America, suggests that there are two Americas in terms of oral health: those who have excellent oral health, and those who are unable to access care and have disparate amounts of dental disease. Since the majority of dental schools are located in urban settings, dental educators need to establish, fund, and sustain outreach programs while sensitizing students to the needs of the underserved. This paper describes the process that Case Western Reserve University School of Dental Medicine used to develop an outreach program to address the needs of underserved children in Cleveland, Ohio. PMID- 15537606 TI - Social insurance for children? PMID- 15537607 TI - Health spending differentials. PMID- 15537610 TI - Disclosure of errors. PMID- 15537611 TI - For-profit accountability. PMID- 15537612 TI - CareFirst conversion. PMID- 15537614 TI - Nurses' working hours. PMID- 15537615 TI - Views of public health. PMID- 15537616 TI - Reducing drug prices. PMID- 15537618 TI - Measuring diabetes management. PMID- 15537621 TI - Lowering diabetes costs. PMID- 15537622 TI - Don't blame the physicians. PMID- 15537623 TI - Long-term care choices. PMID- 15537624 TI - Health spending by age. PMID- 15537630 TI - Insights into the mRNA cleavage mechanism by MazF, an mRNA interferase. AB - MazF is an Escherichia coli toxin that is highly conserved among the prokaryotes and plays an important role in growth regulation. When MazF is induced, protein synthesis is effectively inhibited. However, the mechanism of MazF action has been controversial. Here we unequivocally demonstrate that MazF is an endoribonuclease that specifically cleaves mRNAs at ACA sequences. We then demonstrate its enzymatic specificity using short RNA substrates. MazF cleaves RNA at the 5'-end of ACA sequences, yielding a 2',3'-cyclic phosphate at one side and a free 5'-OH group at the other. Using DNA-RNA chimeric substrates containing XACA, the 2'-OH group of residue X was found absolutely essential for MazF cleavage, whereas all the other residues may be deoxyriboses. Therefore, MazF exhibits exquisite site specificity and has utility as an RNA-restriction enzyme for RNA structural studies or as an mRNA interferase to regulate cell growth in prokaryotic and eukaryotic cells. PMID- 15537629 TI - A sequence within the first transmembrane domain of PEN-2 is critical for PEN-2 mediated endoproteolysis of presenilin 1. AB - Macromolecular complexes containing presenilins (PS), nicastrin (NCT), APH-1, and PEN-2 mediate the gamma-secretase cleavage of the beta-amyloid precursor protein and Notch. APH-1 and NCT stabilize the PS1 holoprotein, whereas PEN-2 is critical for endoproteolysis of PS1. To define the structural domains of PEN-2 that are necessary for mediating PS1 endoproteolysis and gamma-secretase activity, we coexpressed APH-1, NCT, and PS1 together with a series of PEN-2 mutants, which harbored deletions in hydrophilic segments, or chimeric PEN-2 molecules that contained heterologous transmembrane domains (TMDs). We now report that with the exception of the PEN-2 variants with deletions proximal to the TMDs, the vast majority of the deletion variants were functional. Mutants that were nonfunctional were also unstable but were rescued by transposition of a heterologous sequence containing conservative amino acid substitutions into the deleted region. Notably, the carboxyl-terminal hydrophilic domain of PEN-2 was dispensable for promoting PS1 endoproteolysis but was critical for stabilizing the resulting PS1 derivatives. More importantly, we demonstrated that a chimeric PEN-2 with a replacement of the TMD2 with the TMD1 from sterol regulatory element binding protein 1 (SREBP-1) is fully functional but that a chimeric PEN-2 with a replacement of the TMD1 with the TMD2 from SREBP-1 is not. The function of this latter chimera was rescued by the replacement of the proximal two-thirds of the SREBP-1 TMD2 with the proximal two-thirds of the authentic TMD1 from PEN-2. These results suggest that the proximal two-thirds of the PEN-2 TMD1 is functionally important for endoproteolysis of PS1 holoproteins and the generation of PS1 fragments, essential components of the gamma-secretase complex. PMID- 15537631 TI - DNA elongation by the human DNA polymerase lambda polymerase and terminal transferase activities are differentially coordinated by proliferating cell nuclear antigen and replication protein A. AB - DNA polymerase lambda contains template-dependent (DNA polymerase) and template independent (terminal transferase) activities. In this study we enzymologically characterized the terminal transferase activity of polymerase lambda (pol lambda tdt). Pol lambda-tdt activity was strongly influenced by the nature of the 3' terminal sequence of the DNA substrate, and it required a single-stranded (ss) DNA 3'-overhang of about 9-12 nucleotides for optimal activity. The strong preference observed for pyrimidine versus purine nucleotide incorporation was found to be due, at least partially, to a steric block imposed by the residue Tyr 505 in the active site of pol lambda. Pol lambda-tdt was found to be able to elongate a 3'-ssDNA end by two alternative mechanisms: first, a template independent one resulting in addition of 1 or 2 nucleotides, and second, a template-dependent one where a homopolymeric tract as short as 3 nucleotides at the 3'-end could be used as a template to direct DNA polymerization by a looping back mechanism. Furthermore repetitive cycles of DNA synthesis resulted in the expansion of such a short homopolymeric terminal sequence. Most importantly we found that the proliferating cell nuclear antigen was able to selectively block the looping back mechanism while stimulating the single terminal nucleotide addition. Finally replication protein A completely suppressed the transferase activity of pol lambda while stimulating the polymerase activity, suggesting that proliferating cell nuclear antigen and replication protein A can coordinate the polymerase and the terminal transferase activities of pol lambda. PMID- 15537632 TI - Mutations in the spacer region of Drosophila mitochondrial DNA polymerase affect DNA binding, processivity, and the balance between Pol and Exo function. AB - The catalytic subunit (alpha) of mitochondrial DNA polymerase (pol gamma) shares conserved DNA polymerase and 3'-5' exonuclease active site motifs with Escherichia coli DNA polymerase I and bacteriophage T7 DNA polymerase. A major difference between the prokaryotic and mitochondrial proteins is the size and sequence of the region between the exonuclease and DNA polymerase domains, referred to as the spacer in pol gamma-alpha. Four gamma-specific conserved sequence elements are located within the spacer region of the catalytic subunit in eukaryotic species from yeast to humans. To elucidate the functional roles of the spacer region, we pursued deletion and site-directed mutagenesis of Drosophila pol gamma. Mutant proteins were expressed from baculovirus constructs in insect cells, purified to near homogeneity, and analyzed biochemically. We find that mutations in three of the four conserved sequence elements within the spacer alter enzyme activity, processivity, and/or DNA binding affinity. In addition, several mutations affect differentially DNA polymerase and exonuclease activity and/or functional interactions with mitochondrial single-stranded DNA binding protein. Based on these results and crystallographic evidence showing that the template-primer binds in a cleft between the exonuclease and DNA polymerase domains in family A DNA polymerases, we propose that conserved sequences within the spacer of pol gamma may position the substrate with respect to the enzyme catalytic domains. PMID- 15537633 TI - Drosophila melanogaster topoisomerase IIIalpha preferentially relaxes a positively or negatively supercoiled bubble substrate and is essential during development. AB - Eukaryotic type IA topoisomerases are important for the normal function of the cell, and in some cases essential for the organism, although their role in DNA metabolism remains to be elucidated. In this study, we cloned Drosophila melanogaster topoisomerase (topo) IIIalpha from an embryonic cDNA library and expressed and purified the protein to >95% homogeneity. This enzyme partially relaxes a hypernegatively supercoiled plasmid substrate consistent with other purified topo IIIs. A novel, covalently closed bubble substrate was prepared for this study, which topo IIIalpha fully relaxed, regardless of the handedness of the supercoils. Experiments with the bubble substrate demonstrate that topo IIIalpha has much different reaction preferences from those obtained by plasmid substrate-based assays. This is presumably due to the fact that solution conditions can affect the structure of plasmid based substrates and therefore their suitability as a substrate. A mutant allele of the Top3alpha gene, Top3alpha191, was isolated through imprecise excision mutagenesis of an existing P-element inserted in the first intron of the gene. Top3alpha191 is recessive lethal, with most of the homozygous individuals surviving to pupation but never emerging to adulthood. Whereas this mutation can be rescued by a Top3alpha transgene, ubiquitous overexpression of D. melanogaster topo IIIbeta cannot rescue this allele. PMID- 15537634 TI - Depolarization activates ERK and proline-rich tyrosine kinase 2 (PYK2) independently in different cellular compartments in hippocampal slices. AB - In the hippocampus, extracellular signal-regulated kinase (ERK) and the non receptor protein proline-rich tyrosine kinase 2 (PYK2) are activated by depolarization and involved in synaptic plasticity. Both are also activated under pathological conditions following ischemia, convulsions, or electroconvulsive shock. Although in non-neuronal cells PYK2 activates ERK through the recruitment of Src-family kinases (SFKs), the link between these pathways in the hippocampus is not known. We addressed this question using K(+)-depolarized rat hippocampal slices. Depolarization increased the phosphorylation of PYK2, SFKs, and ERK. These effects resulted from Ca(2+) influx through voltage-gated Ca(2+) channels and were diminished by GF109203X, a protein kinase C inhibitor. Inhibition of SFKs with PP2 decreased PYK2 tyrosine phosphorylation dramatically, but not its autophosphorylation on Tyr-402. Moreover, PYK2 autophosphorylation and total tyrosine phosphorylation were profoundly altered in fyn-/- mice, revealing an important functional relationship between Fyn and PYK2 in the hippocampus. In contrast, ERK activation was unaltered by PP2, Fyn knock-out, or LY294002, a phosphatidyl-inositol-3-kinase inhibitor. ERK activation was prevented by MEK inhibitors that had no effect on PYK2. Immunofluorescence of hippocampal slices showed that PYK2 and ERK were activated in distinct cellular compartments in somatodendritic regions and nerve terminals, respectively, with virtually no overlap. Activation of ERK was critical for the rephosphorylation of a synaptic vesicle protein, synapsin I, following depolarization, underlining its functional importance in nerve terminals. Thus, in hippocampal slices, in contrast to cell lines, depolarization-induced activation of non-receptor tyrosine kinases and ERK occurs independently in distinct cellular compartments in which they appear to have different functional roles. PMID- 15537635 TI - Nuclear calpain regulates Ca2+-dependent signaling via proteolysis of nuclear Ca2+/calmodulin-dependent protein kinase type IV in cultured neurons. AB - Accumulating evidence indicates that calpains can reside in or translocate to the cell nucleus, but their functions in this compartment remain poorly understood. Dissociated cultures of cerebellar granule cells (GCs) demonstrate improved long term survival when their growth medium is supplemented with depolarizing agents that stimulate Ca(2+) influx and activate calmodulin-dependent signaling cascades, notably 20 mm KCl. We previously observed Ca(2+)-dependent down regulation of Ca(2+)/calmodulin-dependent protein kinase (CaMK) type IV, which was attenuated by calpain inhibitors, in GCs supplemented with 20 mm KCl (Tremper Wells, B., Mathur, A., Beaman-Hall, C. M., and Vallano, M. L. (2002) J. Neurochem. 81, 314-324). CaMKIV is highly enriched in the nucleus and thought to be critical for improved survival. Here, we demonstrate by immunolocalization/confocal microscopy and subcellular fractionation that the regulatory and catalytic subunits of m-calpain are enriched in GC nuclei, including GCs grown in medium containing 5 mm KCl. Calpain-mediated proteolysis of CaMKIV is selective, as several other nuclear and non-nuclear calpain substrates were not degraded under chronic depolarizing culture conditions. Depolarization and Ca(2+)-dependent down-regulation of CaMKIV were associated with significant alterations in other components of the Ca(2+)-CaMKIV signaling cascade: the ratio of phosphorylated to total cAMP response element-binding protein (a downstream CaMKIV substrate) was reduced by approximately 10-fold, and the amount of CaMK kinase (an upstream activator of CaMKIV) protein and mRNA was significantly reduced. We hypothesize that calpain-mediated CaMKIV proteolysis is an autoregulatory feedback response to sustained activation of a Ca(2+)-CaMKIV signaling pathway, resulting from growth of cultures in medium containing 25 mm KCl. This study establishes nuclear m-calpain as a regulator of CaMKIV and associated signaling molecules under conditions of sustained Ca(2+) influx. PMID- 15537636 TI - Differential effects of modification of membrane cholesterol and sphingolipids on the conformation, function, and trafficking of the G protein-coupled cholecystokinin receptor. AB - The lipid microenvironment of receptors can influence their conformation, function, and regulation. Cholecystokinin (CCK)-stimulated signaling is abnormal in some forms of hyperlipidemia, suggesting the possibility of unique sensitivity to its lipid environment. Here we examined the influence of cholesterol and sphingolipids on CCK receptors in model Chinese hamster ovary cell systems having lipid levels modified. Cholesterol was modulated chemically or metabolically, and sphingolipids were modulated using a temperature-sensitive cell line (SPB-1). Receptor conformation was probed with a fluorescent full agonist ligand, Alexa 488-conjugated Gly-[Nle(28,31)]CCK-(26-33), shown previously to decrease in anisotropy and lifetime when occupying a receptor in the active conformation (Harikumar, K. G., Pinon, D. L., Wessels, W. S., Prendergast, F. G., and Miller, L. J. (2002) J. Biol. Chem. 277, 18552-18560). Anisotropy and lifetime of this probe were increased and prolonged with cholesterol enrichment, and decreased and shortened with depletion of cholesterol or sphingolipids. The increase in these parameters with cholesterol enrichment may reflect change in CCK receptor conformation toward its inactive, uncoupled state. Indeed, cholesterol enrichment resulted in nonproductive agonist ligand binding, with affinity of binding higher than normal and calcium signaling in response to this reduced. In cholesterol- and sphingolipid-depleted states, the receptor moved into conformations that were less than optimal. With cholesterol depletion, both ligand binding and signaling were decreased, yet internalization and trafficking were unperturbed. With sphingolipid depletion, ligand binding and signaling were normal, but internalization and trafficking were markedly inhibited. Of note, normal transferrin receptor trafficking through the same clathrin-dependent pathway was maintained under these conditions. Thus, lipid microenvironment of the CCK receptor is particularly important, with different lipids having distinct effects. PMID- 15537637 TI - The loss of glypican-3 induces alterations in Wnt signaling. AB - Loss-of-function mutations of the GPC3 gene are the cause of the human Simpson Golabi-Behmel syndrome. Based on the overgrowth phenotype of the Simpson-Golabi Behmel syndrome patients and the key role played by the insulin-like growth factor (IGF) signaling system in regulating embryonic growth, it was speculated that GPC3 regulates IGF signaling. In order to test the validity of this hypothesis, we mated GPC3 knockout mice with insulin receptor substrate-1 (IRS-1) nullizygous mice. We found that GPC3 regulates organism growth independent of IRS 1, suggesting that GPC3 does not modulate IGF signaling. Instead, we found that GPC3 knockout mice exhibit alterations in the Wnt signaling pathway, which is also associated with the regulation of cell proliferation. In particular, the loss of GPC3 led to the inhibition of the non-canonical Wnt/JNK signaling pathway, while concomitantly causing the activation of canonical Wnt/beta-catenin signaling. These in vivo findings were confirmed in vitro upon the ectopic overexpression of GPC3 in mesothelioma cells. In these cells, the GPC3-induced increase in JNK activity was associated with an enhanced response to Wnt5a. Most interestingly, the heparan sulfate chains of GPC3 were not required for its stimulatory activity on Wnt5a signaling and for the formation of GPC3-Wnt5a complexes. We propose that at least in some cell types GPC3 serves as a selective regulator of Wnt signaling, by potentiating non-canonical Wnt signaling, while inhibiting the canonical Wnt signaling pathway. PMID- 15537638 TI - Destabilization of the transmembrane domain induces misfolding in a phenotypic mutant of cystic fibrosis transmembrane conductance regulator. AB - Two phenotypic missense mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) channel pore (L346P and R347P in transmembrane (TM) segment 6) involve gain of a proline residue, but only L346P represents a significant loss of segment hydropathy. We show here that, for synthetic peptides corresponding to sequences of CFTR TM6 segments, circular dichroism spectra of wild type and R347P TM6 in membrane mimetic environments are virtually identical, but L346P loses approximately 50% helicity, implying a membrane insertion defect in the latter mutant. A similar defect was observed in the corresponding double spanning ("hairpin") TM5/6-L346P synthetic peptide. Examination of the biogenesis of CFTR revealed that the full-length protein harboring the L346P mutation is rapidly degraded at the endoplasmic reticulum (ER), whereas the wild type and the R347P protein process normally. Furthermore, a second site mutation (R347I) that restores in vitro membrane insertion and folding of the TM5/6-L346P peptide also rescues the folding and cell surface chloride channel function of full-length L346P CFTR. The correlated in vitro/in vivo results demonstrate that destabilizing local hydrophobic character represents a sufficient signal for marking CFTR as a non-native protein by the ER quality control, with accompanying deleterious consequences to global protein folding events. PMID- 15537639 TI - Fluctuations of intracellular iron modulate elastin production. AB - Production of insoluble elastin, the major component of elastic fibers, can be modulated by numerous intrinsic and exogenous factors. Because patients with hemolytic disorders characterized with fluctuations in iron concentration demonstrate defective elastic fibers, we speculated that iron might also modulate elastogenesis. In the present report we demonstrate that treatment of cultured human skin fibroblasts with low concentration of iron 2-20 microm (ferric ammonium citrate) induced a significant increase in the synthesis of tropoelastin and deposition of insoluble elastin. Northern blot and real-time reverse transcription-PCR analysis revealed that treatment with 20 microm iron led to an increase of approximately 3-fold in elastin mRNA levels. Because treatment with an intracellular iron chelator, desferrioxamine, caused a significant decrease in elastin mRNA level and consequent inhibition of elastin deposition, we conclude that iron facilitates elastin gene expression. Our experimental evidence also demonstrates the existence of an opposite effect, in which higher, but not cytotoxic concentrations of iron (100-400 microm) induced the production of intracellular reactive oxygen species that coincided with a significant decrease in elastin message stability and the disappearance of iron-dependent stimulatory effect on elastogenesis. This stimulatory elastogenic effect was reversed, however, in cultures simultaneously treated with high iron concentration (200 microm) and the intracellular hydroxyl radical scavenger, dimethylthiourea. Thus, presented data, for the first time, demonstrate the existence of two opposite iron-dependent mechanisms that may affect the steady state of elastin message. We speculate that extreme fluctuations in intracellular iron levels result in impaired elastic fiber production as observed in hemolytic diseases. PMID- 15537641 TI - Porcine dentin sialoprotein is a proteoglycan with glycosaminoglycan chains containing chondroitin 6-sulfate. AB - Dentin sialoprotein (DSP) is a glycoprotein that is critical for proper tooth dentin formation, but little is known about the nature of its carbohydrate attachments and other post-translational modifications. We have isolated DSP from pig dentin and demonstrate that it is a proteoglycan. Polyclonal antibodies were raised in chicken against recombinant pig DSP, and used to identify native DSP in fractions of tooth dentin proteins extracted from developing pig molars. Amino acid analyses and characterization of lysylendopeptidase cleavage products confirmed that the purified protein was DSP, and that Arg391 is at the DSP C terminus. On SDS-PAGE and on urea gels, DSP appeared as a smear extending from 280 to 100 kDa, but in the presence of beta-mercaptoethanol the top of the DSP smear disappeared. The high molecular weight material was likely comprised of covalent DSP dimers connected by a disulfide bridge at Cys205. Oligosaccharides were released from DSP following N- and O-linked glycosidase digestions, but these digestions had little effect on the apparent molecular weight of DSP on SDS PAGE, when compared with the significant reduction following chondroitinase ABC digestion. Glycosaminoglycanases with assorted glycosaminoglycan (GAG) cleavage specificities coupled with Western analyses of the cleaved GAG "stubs" demonstrated that the DSP GAG attachments contain chondroitin 6-sulfate, but not keratan sulfate, heparan sulfate, chondroitin, or chondroitin 4-sulfate. DSP binds biotin-labeled hyaluronic acid, and such binding is inhibited by the addition of unlabeled hyaluronic acid. We conclude that DSP is a proteoglycan and that GAG attachments are the predominant structural feature of porcine DSP. PMID- 15537640 TI - Spectroscopic and redox properties of a CooA homologue from Carboxydothermus hydrogenoformans. AB - CooA is a CO-sensing transcriptional activator that contains a b-type heme as the active site for sensing its physiological effector, CO. In this study, the spectroscopic and redox properties of a new CooA homologue from Carboxydothermus hydrogenoformans (Ch-CooA) were studied. Spectroscopic and mutagenesis studies revealed that His-82 and the N-terminal alpha-amino group were the axial ligands of the Fe(III) and Fe(II) hemes in Ch-CooA and that the N-terminal alpha-amino group was replaced by CO upon CO binding. Two neutral ligands, His-82 and the N terminal alpha-amino group, are coordinated to the Fe(III) heme in Ch-CooA, whereas two negatively charged ligands, a thiolate from Cys-75 and the nitrogen atom of the N-terminal Pro, are the axial ligands of the Fe(III) heme in Rr-CooA. The difference in the coordination structure of the Fe(III) heme resulted in a large positive shift of redox potentials of Ch-CooA compared with Rr-CooA. Comparing the properties of Ch-CooA and Rr-CooA demonstrates that the essential elements for CooA function will be: (i) the heme is six-coordinate in the Fe(III), Fe(II), and Fe(II)-CO forms; (ii) the N-terminal is coordinated to the heme as an axial ligand, and (iii) CO replaces the N-terminal bound to the heme upon CO binding. PMID- 15537642 TI - Translational mobility of the type 3 inositol 1,4,5-trisphosphate receptor Ca2+ release channel in endoplasmic reticulum membrane. AB - The inositol 1,4,5-trisphosphate receptor (InsP3R) is an integral membrane protein in the endoplasmic reticulum (ER) which functions as a ligand-gated Ca2+ release channel. InsP3-mediated Ca2+ release modulates the cytoplasmic free Ca2+ concentration ([Ca2+]i), providing a ubiquitous intracellular signal with high temporal and spatial specificity. Precise localization of the InsP3R is believed to be important for providing local [Ca2+] regulation and for ensuring efficient functional coupling between Ca2+ release sites by enabling graded recruitment of channels with increasing stimulus strength in the face of the intrinsically unstable regenerative process of Ca2+-induced Ca2+ release. Highly localized Ca2+ release has been attributed to the ability of the InsP3R channels to cluster and to be localized to discrete areas, suggesting that mechanisms may exist to restrict their movement. Here, we examined the lateral mobility of the type 3 isoform of the InsP3R (InsP3R3) in the ER membrane by performing confocal fluorescence recovery after photobleaching of an InsP3R3 with green fluorescent protein fused to its N terminus. In Chinese hamster ovary and COS-7 cells, the diffusion coefficient D was approximately 4 x 10(-10) cm2/s at room temperature, a value similar to that determined for other ER-localized integral membrane proteins, with a high fraction (approximately 75%) of channels mobile. D was modestly increased at 37 degrees C, and it as well as the mobile fraction were reversibly reduced by ATP depletion. Although disruption of the actin cytoskeleton (latrunculin) was without effect, disruption of microtubules (nocodazole) reduced D by half without affecting the mobile fraction. We conclude that the entire ER is continuous in these cells, with the large majority of InsP3R3 channels free to diffuse throughout it, at rates that are comparable with those measured for other polytopic ER integral membrane proteins. The observed InsP3R3 mobility may be higher than its intrinsic diffusional mobility because of additional ATP- and microtubule-facilitated motility of the channel. PMID- 15537643 TI - The transcription factor NFAT3 mediates neuronal survival. AB - Neuronal apoptosis is critical for normal development of the mammalian nervous system and also contributes to the pathogenesis of ischemic and degenerative diseases of the brain. Apoptosis of neurons is tightly regulated by extrinsic signals including growth factors and neuronal activity, but the intracellular mechanisms by which these signals promote neuronal survival are incompletely understood. We report that the transcription factor NFAT3 plays a critical role in mediating survival of granule neurons of the developing cerebellum. NFAT3 accumulated in the nucleus of primary granule neurons under survival conditions of serum growth factors and neuronal activity that was elicited by depolarization with high K(+). In contrast, deprivation of serum and K(+), which leads to neuronal apoptosis, triggered NFAT3 nuclear export. Treatment of granule neurons with Li(+), an inhibitor of the NFAT export kinase GSK3, prevented the nuclear export of NFAT3 and increased granule cell survival even under pro-apoptotic conditions. Thus, the nuclear localization of NFAT3 correlated tightly with granule neuron survival. Consistent with a pro-survival function for NFAT3, genetic knockdown of NFAT3 by RNA interference in primary granule neurons led to increased apoptosis even in neurons cultured under survival conditions. Conversely, expression of a constitutively active form of NFAT protected neurons against apoptosis induced by serum withdrawal and low K(+). Taken together, these results reveal an essential function for NFAT3-mediated transcription in neuronal survival that may play important roles in the developing and mature brain. PMID- 15537644 TI - Reconstitution of regulated phosphorylation of FcepsilonRI by a lipid raft excluded protein-tyrosine phosphatase. AB - To examine the exquisite regulation of IgE-FcepsilonRI tyrosine phosphorylation by Lyn kinase that is stimulated by antigen-mediated cross-linking, we utilized co-expression of FcepsilonRI and Lyn in Chinese hamster ovary cells, which results in high basal levels of Lyn kinase activity and spontaneous phosphorylation of FcepsilonRI. We found that co-expression of a lipid raft excluded transmembrane tyrosine phosphatase, PTPalpha, suppresses Lyn kinase activity and markedly reduces the level of spontaneous phosphorylation of FcepsilonRI, while facilitating its antigen-stimulated phosphorylation. Other tyrosine phosphatases, including SHP-1, CD45, and a lipid raft-preferring chimeric version of PTPalpha fail to reconstitute antigen-dependent FcepsilonRI phosphorylation. We concluded that both substrate specificity and submembrane location are critical to phosphatase-mediated regulation of Lyn kinase activity that supports activation of FcepsilonRI. PMID- 15537645 TI - Analysis of HIV-1 viral infectivity factor-mediated proteasome-dependent depletion of APOBEC3G: correlating function and subcellular localization. AB - To study how HIV-1 viral infectivity factor (Vif) mediates proteasome-dependent depletion of host factor APOBEC3G, functional and nonfunctional Vif-APOBEC3G interactions were correlated with subcellular localization. APOBEC3G localized throughout the cytoplasm and co-localized with gamma-tubulin, 20 S proteasome subunit, and ubiquitin at punctate cytoplasmic bodies that can be used to monitor the Vif-APOBEC3G interaction in the cell. Through immunostaining and live imaging, we showed that a substantial fraction of Vif localized to the nucleus, and this localization was impaired by deletion of amino acids 12-23. When co expressed, Vif exhibited more pronounced localization to the cytoplasm and reduced the total cellular levels of APOBEC3G but rarely co-localized with APOBEC3G at cytoplasmic bodies. On the contrary, Vif(C114S), which is inactive but continues to interact with APOBEC3G, stably associated with APOBEC3G in the cytoplasm, resulting in complete co-localization at cytoplasmic bodies and a dose dependent exclusion of Vif(C114S) from the nucleus. Following proteasome inhibition, cytoplasmic APOBEC3G levels increased, and both proteins co accumulated nonspecifically into a vimentin-encaged aggresome. Furthermore in the presence or absence of APOBEC3G, Vif localization was significantly altered by proteasome inhibition, suggesting that aberrant localization may also contribute to the loss of Vif function. Finally mutations at Vif Ile(9) disrupted the ability of Vif or Vif(C114S) to coimmunoprecipitate and to co-localize with APOBEC3G, suggesting that the N terminus of Vif mediates interactions with APOBEC3G. Taken together, these results demonstrate that cytoplasmic Vif-APOBEC3G interactions are required but are not sufficient for Vif to modulate APOBEC3G and can be monitored by co-localization in vivo. PMID- 15537646 TI - Exploring strategies for the design of artificial transcription factors: targeting sites proximal to known regulatory regions for the induction of gamma globin expression and the treatment of sickle cell disease. AB - Artificial transcription factors can be engineered to interact with specific DNA sequences to modulate endogenous gene expression within cells. A significant hurdle to implementation of this approach is the selection of the appropriate DNA sequence for targeting. We reasoned that a good target site should be located in chromatin, where it is accessible to DNA-binding proteins, and it should be in the close vicinity of known transcriptional regulators of the gene. Here we have explored the efficacy of these criteria to guide our selection of potential regulators of gamma-globin expression. Several zinc finger-based transcriptional activators were designed to target the sites proximal to the -117-position of the gamma-globin promoter. This region is proximal to the binding sites of known and potential natural transcription factors. Design and study of three transcription factors identified the potent transcriptional activator, gg1-VP64-HA. This transcription factor was able to interact directly with the gamma-globin promoter and up-regulate expression of reporter gene constructs as well as the endogenous gene in a selective manner. Transfection of a gg1-VP64-HA expression vector or retroviral delivery of this transcription factor into the erythroleukemia cell line K562 resulted in an increase of fetal hemoglobin. The gamma-globin content of cells expressing gg1-vp64-HA showed up to 16-fold higher levels of fetal hemoglobin than the native K562 cell line. These transcriptional activators constitute a novel class of regulators of the globin locus that may be suitable for treatment of diseases arising from mutations in this locus such as sickle cell disease and thalassemic diseases. PMID- 15537647 TI - Glucocorticoids inhibit the transcriptional activity of LEF/TCF in differentiating osteoblasts in a glycogen synthase kinase-3beta-dependent and independent manner. AB - Glucocorticoids, widely used as immune suppressors, cause osteoporosis by inhibiting bone formation. In MC3T3-E1 osteoblast-like cultures, dexamethasone (DEX) activates glycogen synthase kinase-3beta (GSK3beta) and inhibits a differentiation-related cell cycle that occurs at a commitment stage immediately after confluence. Here we show that DEX inhibition of the differentiation-related cell cycle is associated with a decrease in beta-catenin levels and inhibition of LEF/TCF-mediated transcription. These inhibitory activities are no longer observed in the presence of lithium, a GSK3beta inhibitor. DEX decreased the serum-responsive phosphorylation of protein kinase B/Akt-Ser(473) within minutes, and this inhibition was also observed after 12 h. When the phosphatidylinositol 3 kinase (PI3K)/Akt pathway was inhibited by wortmannin, DEX no longer inhibited beta-catenin levels. Furthermore, DEX-mediated inhibition of LEF/TCF transcriptional activity was attenuated in the presence of dominant negative forms of either PI3K or protein kinase B/Akt. These results suggest cross-talk between the PI3K/Akt and Wnt signaling pathways. Consistent with a role for Wnt signaling in the osteoblast differentiation-related cell cycle, wortmannin partially negated the DEX inhibition of this cell cycle. DEX also induced histone deacetylase (HDAC) 1, which is known to inhibit LEF/TCF transcriptional activity. Overexpression of HDAC1 negated the inhibitory effect of DEX on LEF/TCF transcriptional activity. In the presence of trichostatin A, a deacetylase inhibitor, DEX-mediated inhibition of the differentiation-related cell cycle was partially negated. When administered together, wortmannin and trichostatin A completely negated the inhibitory effect of DEX on the differentiation-related cell cycle. These results suggest that inhibition of a PI3K/Akt/GSK3beta/beta catenin/LEF axis and stimulation of HDAC1 cooperate to mediate the inhibitory effect of DEX on Wnt signaling and the osteoblast differentiation-related cell cycle. PMID- 15537648 TI - Orexin-A-induced Ca2+ entry: evidence for involvement of trpc channels and protein kinase C regulation. AB - The orexins are peptide transmitters/hormones, which exert stimulatory actions in many types of cells via the G-protein-coupled OX(1) and OX(2) receptors. Our previous results have suggested that low (subnanomolar) concentrations of orexin A activate Ca(2+) entry, whereas higher concentrations activate phospholipase C, Ca(2+) release, and capacitative Ca(2+) entry. As shown here, the Ca(2+) response to subnanomolar orexin-A concentrations was blocked by activation of protein kinase C by using different approaches (12-O-tetradecanoylphorbol acetate, dioctanoylglycerol, and diacylglycerol kinase inhibition) and protein phosphatase inhibition by calyculin A. The Ca(2+) response to subnanomolar orexin-A concentrations was also blocked by Mg(2+), dextromethorphan, and tetraethylammonium. These treatments neither affected the response to high concentrations of orexin-A nor the thapsigargin-stimulated capacitative entry. The capacitative entry was instead strongly suppressed by SKF96365. An inward membrane current activated by subnanomolar concentrations of orexin-A and the currents activated upon transient expression of trpc3 channels were also sensitive to Mg(2+), dextromethorphan, and tetraethylammonium. Responses to subnanomolar concentrations of orexin-A (Ca(2+) elevation, inward current, and membrane depolarization) were voltage-dependent with a loss of the response around -15 mV. By using reverse transcription-PCR, mRNA for the trpc1-4 channel isoforms were detected in the CHO-hOX1-C1 cells. The expression of truncated TRPC channel isoforms, in particular trpc1 and trpc3, reduced the response to subnanomolar concentrations of orexin-A but did not affect the response to higher concentrations of orexin-A. The results suggest that activation of the OX(1) receptor leads to opening of a Ca(2+)-permeable channel, involving trpc1 and -3, which is controlled by protein kinase C. PMID- 15537649 TI - Identification of endoglin in rat hepatic stellate cells: new insights into transforming growth factor beta receptor signaling. AB - Transforming growth factor beta (TGF-beta) signaling is mediated by the cell surface TGF-beta type I (ALK5), type II, and the accessory type III receptors endoglin and betaglycan. Hepatic stellate cells (HSC), the most profibrogenic cell type in the liver, express ALK5, TbetaRII, and betaglycan. To monitor the expression of betaglycan in HSC, we used the commercially available antibody sc 6199 in Western blot analysis. This antibody, raised against a peptide mapping at the carboxyl terminus of the human betaglycan, is claimed to be specific for betaglycan, although it is known that the C-terminal domain is highly conserved in type III receptors. Proteins recognized in HSC by sc-6199 did not match the characteristic migration pattern of betaglycan. Moreover, the determined molecular weight (M(r) 160) and the observed reductant sensitivity after treatment with dithiothreitol resemble those of a closely related type III receptor, endoglin (CD105). Endoglin, a disulfide-linked homodimer, is an accessory component of the TGF-beta receptor complex and mainly expressed on endothelial cells. The presence of endoglin in HSC of rat liver was confirmed by molecular cloning of the endoglin cDNA and immunocytochemistry. The reactivity of sc-6199 with both auxiliary TGF-beta receptors (betaglycan and endoglin) from rats was demonstrated by Western blot and immunocytochemical analysis of cells heterologously expressing these proteins. Furthermore, Northern and Western blotting revealed that both betaglycan and endoglin genes are differentially regulated in HSC and in transdifferentiated myofibroblasts (MFB). By surface labeling and immunoprecipitation experiments, we show that endoglin is found in significant amounts exposed at the plasma membrane of HSC and MFB, which is a pivotal prerequisite for binding of and signaling in response to TGF-beta. In conclusion, we hypothesize that TGF-beta signals in HSC and MFB are tuned by two different interconnected signaling pathways, as it was previously demonstrated for endothelial cells. PMID- 15537650 TI - Glycan-independent role of calnexin in the intracellular retention of Charcot Marie-tooth 1A Gas3/PMP22 mutants. AB - Missense point mutations in Gas3/PMP22 are responsible for the peripheral neuropathies Charcot-Marie-Tooth 1A and Dejerine Sottas syndrome. These mutations induce protein misfolding with the consequent accumulation of the proteins in the endoplasmic reticulum and the formation of aggresomes. During folding, Gas3/PMP22 associates with the lectin chaperone calnexin. Here, we show that calnexin interacts with the misfolded transmembrane domains of Gas3/PMP22, fused to green fluorescent protein, in a glycan-independent manner. In addition, photobleaching experiments in living cells revealed that Gas3/PMP22-green fluorescent protein mutants are mobile but diffuse at almost half the diffusion coefficient of wild type protein. Our results support emerging models for a glycan-independent chaperone role for calnexin and for the mechanism of retention of misfolded membrane proteins in the endoplasmic reticulum. PMID- 15537651 TI - Trypanothione synthesis in crithidia revisited. AB - In Crithidia fasciculata the biosynthesis of trypanothione (N(1),N(8) bis(glutathionyl)spermidine; reduced trypanothione), a redox mediator unique to and essential for pathogenic trypanosomatids, was assumed to be achieved by two distinct enzymes, glutathionylspermidine synthetase and trypanothione synthetase (TryS), and only the first one was adequately characterized. We here report that the TryS of C. fasciculata, like that of Trypanosoma species, catalyzes the entire synthesis of trypanothione, whereas its glutathionylspermidine synthetase appears to be specialized for Gsp synthesis. A gene (GenBanktrade mark accession number AY603101) implicated in reduced trypanothione synthesis of C. fasciculata was isolated from genomic DNA and expressed in Escherichia coli as His-tagged or Nus fusion proteins. The expression product proved to be a trypanothione synthetase (Cf-TryS) that also displayed a glutathionylspermidine synthetase, an amidase, and marginal ATPase activity. The dual specificity of the Cf-TryS preparations was not altered by removal of the tags. Steady-state kinetic analysis of Cf-TryS yielded a pattern that was compatible with a concerted substitution mechanism, wherein the enzyme forms a ternary complex with Mg(2+) ATP and GSH to phosphorylate GSH and then ligates the glutathionyl residue to glutathionylspermidine. Limiting K(m) values for GSH, Mg(2+)-ATP, and glutathionylspermidine were 407, 222, and 480 microm, respectively, and the k(cat) was 8.7 s(-1) for the TryS reaction. Mutating Arg-553 or Arg-613 to Lys, Leu, Gln, or Glu resulted in marked reduction or abrogation (R553E) of activity. Limited proteolysis with factor Xa or trypsin resulted in cleavage at Arg-556 that was accompanied by loss of activity. The presence of substrates, in particular of ATP and GSH alone or in combination, delayed proteolysis of wild type Cf-TryS and Cf-TryS R553Q but not in Cf-TryS R613Q, which suggests dynamic interactions of remote domains in substrate binding and catalysis. PMID- 15537652 TI - Regulation of ultraviolet B-induced phosphorylation of histone H3 at serine 10 by Fyn kinase. AB - Ultraviolet B (UVB) induces phosphorylation of histone H3 at serine 10, and mitogen-activated protein kinases are involved in this signal transduction pathway. Here we provide evidence that Fyn kinase, a member of the Src kinase family, is involved in the UVB-induced phosphorylation of histone H3 at serine 10. UVB distinctly increased Fyn kinase activity and phosphorylation. Fyn kinase inhibitors 4-amino-5-(4-chlorophenyl)-7(t-butyl)pyrazol(3,4-d)pyramide and leflunomide, an Src kinase inhibitor, suppressed both UVB-induced phosphorylation of histone H3 at serine 10 and Fyn kinase activity and phosphorylation. UVB induced phosphorylation of histone H3 at serine 10 was blocked by either a dominant-negative mutant of Fyn (DNM-Fyn) kinase or small interfering RNA of Fyn kinase. UVB-induced phosphorylation and activities of ERKs and protein kinase B/Akt were markedly inhibited by DNM-Fyn kinase. However, DNM-Fyn kinase did not inhibit UVB-induced phosphorylation of p38 MAPK or c-Jun N-terminal kinases. Active Fyn kinase phosphorylated histone H3 at serine 10 in vitro, and the phosphorylated Fyn kinase could translocate into the nucleus of HaCaT cells. These results indicate that Fyn kinase plays a key role in the UVB-induced phosphorylation of histone H3 at serine 10. PMID- 15537653 TI - Strong and rapid induction of osteoblast differentiation by Cbfa1/Til-1 overexpression for bone regeneration. AB - Core binding factor alpha-1 (Cbfa1), known as an essential transcription factor for osteogenic lineage, has two major N-terminal isoforms: Pebp2alphaA and Til-1. To study the roles of these isoforms in bone regeneration, we applied an adenoviral vector carrying their genes to transduce primary osteoprogenitor cells in vitro and in vivo. Overexpression of the two isoforms induced rapid and marked osteoblast differentiation, with Til-1 being more effective in vitro, by examination of the alkaline phosphatase activity, calcium content, and Alizarin red staining. Til-1 overexpressing cells/porous ceramic composites were transplanted into subcutaneous and bone defect sites in Fischer rats (cultured bone transplantation model) and markedly affected in vivo bone formation and osteoblast markers. The results demonstrated that the reconstitution of bone tissues, such as cortical bone and trabecular bone was accelerated by implantation of Til-1 overexpressing cells/porous ceramic composites. Moreover, the new bone formation by Til-1 overexpression appeared to reflect replacement of new bone within the implant boundaries. To ascertain whether implanted Cbfa1 overexpressing cells could differentiate into osteogenic cells to create bone or whether it stimulated the surrounding recipient tissue to regenerate bone, implanted male donor cells were visualized by fluorescent in situ hybridization analysis. The proportion of implanted cells in the presumptive bone forming region was over 80% and did not change throughout from 3 days to 8 weeks after implantation. These findings suggested that the newly formed bone in the porous area of the scaffold is mostly produced by the implanted donor cells or their derived cells, effectively by Til-1 overexpression. PMID- 15537654 TI - Insulin receptor substrate-2 proteasomal degradation mediated by a mammalian target of rapamycin (mTOR)-induced negative feedback down-regulates protein kinase B-mediated signaling pathway in beta-cells. AB - Regulation of insulin receptor substrate (IRS)-2 expression is critical to beta cell survival, but the mechanisms that control this are complex and undefined. Here in pancreatic beta-cells (INS-1), chronic exposure (>8 h) to 15 mm glucose and/or 5 nm IGF-1, increased Ser/Thr phosphorylation of IRS-2, which correlated with decreased IRS-2 levels. This glucose/IGF-1-induced decrease in IRS-2 levels was prevented by the proteasomal inhibitor, lactacystin. In addition, the glucose/IGF-1-induced increase in Ser/Thr phosphorylation of IRS-2 and the subsequent decrease in INS-1 cell IRS-2 protein levels was thwarted by the mammalian target of rapamycin(mTOR) inhibitor, rapamycin. Moreover, adenoviral mediated expression of constitutively active mTOR (mTORDelta) further increased glucose/IGF-1-induced Ser/Thr phosphorylation of IRS-2 and decreased IRS-2 protein levels, whereas adenoviral-mediated expression of "kinase-dead" mTOR (mTOR-KD) conversely reduced Ser/Thr phosphorylation of IRS-2 and maintained IRS 2 protein levels. In adenoviral-infected beta-cells expressing mTORDelta, the decrease in IRS-2 protein levels was also prevented by rapamycin or lactacystin, further indicating a proteasomal mediated degradation of IRS-2 mediated via mTOR induced Ser/Thr phosphorylation of IRS-2. Finally, we found that chronic activation of mTOR leading to decreased levels of IRS-2 in INS-1 cells led to a significant decrease in PKB activation and consequently increased beta-cell apoptosis. Thus, chronic activation of mTOR by glucose (and/or IGF-1) in beta cells leads to increased Ser/Thr phosphorylation of IRS-2 that targets it for proteasomal degradation, resulting in decreased IRS-2 expression and increased beta-cell apoptosis. This may be a contributing mechanism as to how beta-cell mass is decreased by chronic hyperglycemia in the pathogenesis of type-2 diabetes. PMID- 15537655 TI - Adipogenic transcriptional regulation of hepatic stellate cells. AB - Hepatic stellate cells (HSC) undergo transdifferentiation (activation) from lipid storing pericytes to myofibroblastic cells to participate in liver fibrogenesis. Our recent work demonstrates that depletion of peroxisome proliferator-activated receptor gamma (PPARgamma) constitutes one of the key molecular events for HSC activation and that ectopic expression of this nuclear receptor achieves the phenotypic reversal of activated HSC to the quiescent cells. The present study extends these findings to test a novel hypothesis that adipogenic transcriptional regulation is required for the maintenance of HSC quiescence. Comparative analysis of quiescent and activated HSC in culture reveals higher expression of putative adipogenic transcription factors such as CCAAT/enhancer-binding protein (C/EBP) alpha, C/EBPbeta, C/EBPdelta, PPARgamma, liver X receptor alpha, sterol regulatory element-binding protein 1c and of adipocyte-specific genes in the quiescent cells. Conversely, activated HSC have increased expression of PPARbeta, a transcription factor known to promote fatty acid oxidation. A treatment of activated HSC with the adipocyte differentiation mixture (isobutylmethylxanthine, dexamethasone, and insulin) or ectopic expression of PPARgamma or SREBP-1c in these cells, induces a panel of adipogenic transcription factors, reduces PPARbeta, and causes the phenotypic reversal to quiescent HSC. These results support the importance of adipogenic transcriptional regulation in HSC quiescence and provide a new framework for identifying novel molecular targets for the treatment of liver cirrhosis. PMID- 15537656 TI - A direct interaction between Cdc42 and vesicle-associated membrane protein 2 regulates SNARE-dependent insulin exocytosis. AB - In pancreatic beta cells, insulin granule exocytosis is regulated by SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor protein (SNAP) receptor) proteins, and this is coupled to cortical F-actin reorganization via the Rho family GTPase Cdc42 by an unknown mechanism. We investigated interactions among the target SNARE protein Syntaxin 1A and the vesicle associated membrane SNARE protein (VAMP2) with Cdc42 and compared these structural interactions with their functional importance to glucose-stimulated insulin secretion in MIN6 beta cells. Subcellular fractionation analyses revealed a parallel redistribution of Cdc42 and VAMP2 from the granule fraction to the plasma membrane in response to glucose that temporally corresponded with the glucose-induced activation of Cdc42. Moreover, within these fractions Cdc42 and VAMP2 were found to co-immunoprecipitate under basal and glucose-stimulated conditions, suggesting that they moved as a complex. Furthermore, VAMP2 bound both GST-Cdc42-GTPgammaS and GST-Cdc42-GDP, indicating that the Cdc42-VAMP2 complex could form under both cytosolic GDP-bound Cdc42 and plasma membrane GTP bound Cdc42 conformational conditions. In vitro binding analyses showed that VAMP2 bound directly to Cdc42 and that a heterotrimeric complex with Syntaxin 1A could also be formed. Deletion analyses of VAMP2 revealed that only the N terminal 28 residues were required for Cdc42 binding. Expression of this 28 residue VAMP2 peptide in MIN6 beta cells resulted in the specific impairment of glucose-stimulated insulin secretion, indicating a functional importance for the Cdc42-VAMP2 interaction. Taken together, these data suggest a mechanism whereby glucose activates Cdc42 to induce the targeting of intracellular Cdc42-VAMP2 insulin granule complexes to Syntaxin 1A at the plasma membrane. PMID- 15537657 TI - Triacylglycerol is synthesized in a specific subclass of caveolae in primary adipocytes. AB - A principal metabolic function of adipocytes is to synthesize triacylglycerol (TG) from exogenous fatty acids. The level of fatty acids has to be tightly controlled in the adipocyte, as they can act as detergents that rapidly dissolve the plasma membrane, causing cell lysis if allowed to accumulate. Fatty acids therefore have to be efficiently converted to TG and stored in the central lipid droplet. We report that in intact primary adipocytes exogenous oleic acid was taken up and directly converted to TG in the plasma membrane, in a novel subclass of caveolae that specifically contains the protein perilipin. Isolated caveolae catalyzed de novo TG synthesis from oleic acid and glycerol 3-phosphate. Electron microscopy revealed the presence of caveolin and perilipin in caveolae and in lipid-laden bulbs in the plasma membrane, and fluorescence microscopy demonstrated colocalization of fatty acids/TG with caveolin and perilipin at the plasma membrane. A second caveolae fraction was isolated, which lacked perilipin and the triacylglycerol synthesizing enzymes. Both caveolae fractions contained caveolin-1 and the insulin receptor. The findings demonstrate that specific subclasses of caveolae carry out specific functions in cell metabolism. In particular, triacylglycerol is synthesized at the site of fatty acid entry in one of these caveolae classes. PMID- 15537658 TI - A major histocompatibility complex-peptide-restricted antibody and t cell receptor molecules recognize their target by distinct binding modes: crystal structure of human leukocyte antigen (HLA)-A1-MAGE-A1 in complex with FAB-HYB3. AB - Antibodies with T cell receptor-like specificity possess a considerable diagnostic and therapeutic potential, but the structural basis of the interaction between an antibody and an histocompatibility antigen has so far not been determined. We present here the crystal structure (at 2.15 A resolution) of the recombinant, affinity-matured human antibody fragment Fab-Hyb3 bound to the tumor associated human leukocyte antigen (HLA)/peptide complex HLA-A1.MAGE-A1. Fab-Hyb3 employs a diagonal docking mode resembling that of T cell receptors. However, other than these natural ligands, the antibody uses only four of its six complementarity-determining regions for direct interactions with the target. It recognizes the C-terminal half of the MAGE-A1 peptide, the HLA-A1 alpha1-helix, and N-terminal residues of the alpha2-helix, accompanied by a large tilting angle between the two types of molecules within the complex. Interestingly, only a single hydrogen bond between a peptide side chain and Fab-Hyb3 contributes to the interaction, but large buried surface areas with pronounced shape complementarity assure high affinity and specificity for MAGE-A1. The HLA-A1.MAGE-A1.antibody structure is discussed in comparison with those of natural ligands recognizing HLA.peptide complexes. PMID- 15537659 TI - Crystal structure of an ATPase-active form of Rad51 homolog from Methanococcus voltae. Insights into potassium dependence. AB - Homologous gene recombination is crucial for the repair of DNA. A superfamily of recombinases facilitate a central strand exchange reaction in the repair process. This reaction is initiated by coating single-stranded DNA (ssDNA) with recombinases in the presence of ATP and Mg(2+) co-factors to form helical nucleoprotein filaments with elevated ATPase and strand invasion activities. At the amino acid sequence level, archaeal RadA and Rad51 and eukaryal Rad51 and meiosis-specific DMC1 form a closely related group of recombinases distinct from bacterial RecA. Unlike the extensively studied Escherichia coli RecA (EcRecA), increasing evidences on yeast and human recombinases imply that their optimal activities are dependent on the presence of a monovalent cation, particularly potassium. Here we present the finding that archaeal RadA from Methanococcus voltae (MvRadA) is a stringent potassium-dependent ATPase, and the crystal structure of this protein in complex with the non-hydrolyzable ATP analog adenosine 5'-(beta,gamma-iminotriphosphate), Mg(2+), and K(+) at 2.4 A resolution. Potassium triggered an in situ conformational change in the ssDNA binding L2 region concerted with incorporation of two potassium ions at the ATPase site in the RadA crystals preformed in K(+)-free medium. Both potassium ions were observed in contact with the gamma-phosphate of the ATP analog, implying a direct role by the monovalent cations in stimulating the ATPase activity. Cross-talk between the ATPase site and the ssDNA-binding L2 region visualized in the MvRadA structure provides an explanation to the co-factor induced allosteric effect on RecA-like recombinases. PMID- 15537660 TI - Allele-dependent similarity between viral and self-peptide presentation by HLA B27 subtypes. AB - Molecular mimicry is discussed as a possible mechanism that may contribute to the development of autoimmune diseases. It could also be involved in the differential association of the human major histocompatibility subtypes HLA-B(*)2705 and HLA B(*)2709 with ankylosing spondylitis. These two subtypes differ only in residue 116 of the heavy chain (Asp in B(*)2705 and His in B(*)2709), but the reason for the differential disease association is not understood. Using x-ray crystallography, we show here that the viral peptide pLMP2 (RRRWRRLTV, derived from latent membrane protein 2 (residues 236-244) of Epstein-Barr virus) is presented by the B(*)2705 and B(*)2709 molecules in two drastically deviating conformations. Extensive structural similarity between pLMP2 and the self-peptide pVIPR (RRKWRRWHL, derived from vasoactive intestinal peptide type 1 receptor (residues 400-408)) is observed only when the peptides are presented by B(*)2705 because of a salt bridge between Arg(5) of both peptides and the subtype-specific heavy chain residue Asp(116). Combined with functional studies using pLMP2/pVIPR cross-reactive cytotoxic T cell lines and clones, together with target cells presenting these peptides or a modified peptide analogue, our results reveal that a pathogen-derived peptide can exhibit major histocompatibility complex class I subtype-dependent, drastically distinct binding modes. Furthermore, the results demonstrate that molecular mimicry between pLMP2 and pVIPR in the HLA-B27 context is an allele-dependent property. PMID- 15537661 TI - An extensive indoor 222Rn/220Rn monitoring in Shillong, India. AB - The behaviour of ubiquitous radon (222Rn), thoron (220Rn) and their progeny in the indoor atmosphere generally reflects a complex interplay between a number of processes, the most important of which are radioactive alpha-decay, ventilation, attachment to aerosols and deposition on surfaces. The present work involved a long-term (1997-2000) passive monitoring of 222Rn and 220Rn in the indoor environment of Shillong, Meghalaya. The north-east region of India being a zone of high seismicity, the indoor radon and thoron map of the region will provide a better insight and a valuable database for any study related to radon and thoron anomalies. PMID- 15537662 TI - Study and presentation of a fast neutron and photon dosemeter for area and criticality monitoring using radiophotoluminescent glass. AB - This paper describes the results of a study performed on a mixed field neutron/gamma (n/gamma) area dosemeter incorporating radiophotoluminescent (RPL) glass detectors. RPL glass is known to be virtually insensitive to neutrons. The aim of the study was therefore to determine the neutron response of a dosemeter designed to combine n/gamma conversion with RPL detection capability. Monte Carlo calculations as well as measurements using monoenergetic beams and isotopic neutron sources showed this response to be constant, to within 30% in terms of H*(10), and independent of neutron energy from 250 keV to 10 MeV. For area monitoring, tests carried out in nuclear facilities (around PuO2 glove box and shipping casks containing PWR, MOX spent fuels or vitrified fission product) demonstrated that dosemeter response was accurate to within 15%, where the gamma component of the mixed n,gamma field remained below 1 MeV. When exposed in the Silene reactor simulating a criticality accident (10(17) fissions-liquid 235U- e.g. 1 Gy neutron and 1 Gy photon), the dosemeter exhibited good correlation with reference values and other measurement technologies (again to within 30%), for both neutron and gamma absorbed dose. PMID- 15537663 TI - Genetic basis for individual variations in pain perception and the development of a chronic pain condition. AB - Pain sensitivity varies substantially among humans. A significant part of the human population develops chronic pain conditions that are characterized by heightened pain sensitivity. We identified three genetic variants (haplotypes) of the gene encoding catecholamine-O-methyltransferase (COMT) that we designated as low pain sensitivity (LPS), average pain sensitivity (APS) and high pain sensitivity (HPS). We show that these haplotypes encompass 96% of the human population, and five combinations of these haplotypes are strongly associated (P=0.0004) with variation in the sensitivity to experimental pain. The presence of even a single LPS haplotype diminishes, by as much as 2.3 times, the risk of developing myogenous temporomandibular joint disorder (TMD), a common musculoskeletal pain condition. The LPS haplotype produces much higher levels of COMT enzymatic activity when compared with the APS or HPS haplotypes. Inhibition of COMT in the rat results in a profound increase in pain sensitivity. Thus, COMT activity substantially influences pain sensitivity, and the three major haplotypes determine COMT activity in humans that inversely correlates with pain sensitivity and the risk of developing TMD. PMID- 15537664 TI - Urogenital and caudal dysgenesis in adrenocortical dysplasia (acd) mice is caused by a splicing mutation in a novel telomeric regulator. AB - Adrenocortical dysplasia (acd) is a spontaneous autosomal recessive mouse mutant with developmental defects in organs derived from the urogenital ridge. In surviving adult mutants, adrenocortical dysplasia and hypofunction are predominant features. Adults are infertile due to lack of mature germ cells, and 50% develop hydronephrosis due to ureteral hyperplasia. We report the identification of a splice donor mutation in a novel gene, which is the mouse ortholog of a newly discovered telomeric regulator. This gene (Acd) has recently been characterized as a novel component of the TRF1 protein complex that controls telomere elongation by telomerase. Characterization of Acd transcripts in mutant animals reveals two abnormal transcripts, consistent with a splicing defect. Expression of a wild-type Acd transgene in acd mutants rescues the observed phenotype. Most mutants die within 1-2 days of life on the original genetic background. Analysis of these mutant embryos reveals variable, yet striking defects in caudal specification, limb patterning and axial skeleton formation. In the tail bud, reduced expression of Wnt3a and Dll1 correlates with phenotypic severity of caudal regression. In the limbs, expression of Fgf8 is expanded in the dorsal-ventral axis of the apical ectodermal ridge and shortened in the anterior-posterior axis, consistent with the observed loss of anterior digits in older embryos. The axial skeleton of mutant embryos shows abnormal vertebral fusions in cervical, lumbar and caudal regions. This is the first report to show that a telomeric regulator is required for proper urogenital ridge differentiation, axial skeleton specification and limb patterning in mice. PMID- 15537665 TI - Digenic inheritance of deafness caused by mutations in genes encoding cadherin 23 and protocadherin 15 in mice and humans. AB - Mutations in genes coding for cadherin 23 and protocadherin 15 cause deafness in both mice and humans. Here, we provide evidence that mutations at these two cadherin loci can interact to cause hearing loss in digenic heterozygotes of both species. Using a classical genetic approach, we generated mice that were heterozygous for both Cdh23 and Pcdh15 mutations on a uniform C57BL/6J background. Significant levels of hearing loss were detected in these mice when compared to age-matched single heterozygous animals or normal controls. Cytoarchitectural defects in the cochlea of digenic heterozygotes, including degeneration of the stereocilia and a base-apex loss of hair cells and spiral ganglion cells, were consistent with the observed age-related hearing loss of these mice beginning with the high frequencies. In humans, we also have obtained evidence for a digenic inheritance of a USH1 phenotype in three unrelated families with mutations in CDH23 and PCDH15. Altogether, our data indicate that CDH23 and PCDH15 play an essential long-term role in maintaining the normal organization of the stereocilia bundle. PMID- 15537666 TI - Distinct patterns of abnormal GNAS imprinting in familial and sporadic pseudohypoparathyroidism type IB. AB - Pseudohypoparathyroidism type IB (PHPIB) is associated with abnormal imprinting of GNAS, the gene encoding the heterotrimeric G protein Gsalpha and other alternative products. The gene contains three differentially methylated regions (DMRs) located upstream of the Gsalpha promoter (from upstream to downstream): the paternally methylated NESP55 promoter region, the maternally methylated NESP antisense (NESPAS)/XLalphas promoter region and the maternally methylated exon 1A region located just upstream of the Gsalpha promoter. We have now performed a detailed analysis of the GNAS methylation profile in 20 unrelated PHPIB probands. Consistent with prior results, all have loss of exon 1A imprinting (a paternal epigenotype on both alleles). All five probands with familial disease had a deletion mutation within the closely linked STX16 gene and a GNAS imprinting defect involving only the exon 1A region. In contrast, the STX16 mutation was absent in all sporadic cases. The majority of these patients had abnormal imprinting of the more upstream regions in addition to the exon 1A imprinting defect, with eight of 15 having a paternal epigenotype on both alleles throughout the GNAS locus. In virtually all cases, the imprinting status of the NESP55 and NESPAS/XLalphas promoters is concordant, suggesting that their imprinting is co regulated, whereas the imprinting of the NESPAS/XLalphas promoter region and XLalphas first exon is not always concordant even though they are closely linked and lie within the same DMR. Familial and sporadic forms of PHPIB have distinct GNAS imprinting patterns that occur through different defects in the imprinting mechanism. PMID- 15537667 TI - Variable and hierarchical size distribution of L1-retroelement-enriched CENP-A clusters within a functional human neocentromere. AB - Human neocentromeres are fully functional centromeres that arise epigenetically from non-centromeric precursor sequences that are devoid of alpha-satellite DNA. Using chromatin immunoprecipitation (ChIP) and BAC-array analysis, we have previously described a 330 kb binding domain for CENP-A (a histone H3 variant that confers centromere-specific nucleosomal property) at the 10q25 neocentromere found on a chromosome 10-derived marker chromosome mardel(10). For the further detailed analysis of the CENP-A-associated chromatin, we have generated a high resolution genomic array consisting of PCR fragments with an average size of 8 kb, providing an approximately 20-fold increment in analytical resolution. ChIP and PCR-array analysis reveals seven distinct CENP-A-binding clusters within the 330 kb domain, demonstrating the interspersion of CENP-A-associated nucleosomal blocks within the neocentromeric chromatin. Independent ChIP-PCR analysis verified this distribution profile and indicated that histone H3-containing nucleosomes directly intervene the CENP-A-binding clusters. The CENP-A-binding clusters are uneven in size, with the central cluster (>50 kb) being significantly larger than the flanking ones (10-30 kb), and the flanking clusters arranged in an interesting hierarchical and symmetrical configuration of alternating larger and smaller sizes around the central cluster. In silico sequence analysis indicates an approximately 2.5-fold increase in the prevalence of L1 retroelements within the CENP-A-binding clusters when compared with the non CENP-A-binding regions. These results provide insight into the possible role of retroelements in determining the positioning of CENP-A binding at human neocentromeres, and that a hierarchical and symmetrical arrangement of CENP-A binding clusters of varying sizes may be an important structural requirement for mammalian kinetochore assembly and/or to provide stability to withstand polar microtubule forces. PMID- 15537668 TI - The hereditary spastic paraplegia protein spastin interacts with the ESCRT-III complex-associated endosomal protein CHMP1B. AB - Pure hereditary spastic paraplegia is characterized by length-dependent degeneration of the distal ends of long axons. Mutations in spastin are the most common cause of the condition. We set out to investigate the function of spastin using a yeast two-hybrid approach to identify interacting proteins. Using full length spastin as bait, we identified CHMP1B, a protein associated with the ESCRT (endosomal sorting complex required for transport)-III complex, as a binding partner. Several different approaches confirmed the physiological relevance of the interaction in mammalian cells. Epitope-tagged CHMP1B and spastin showed clear cytoplasmic co-localization in Cos-7 and PC12 cells. CHMP1B and spastin interacted specifically in vitro and in vivo in beta-lactamase protein fragment complementation assays, and spastin co-immunoprecipitated with CHMP1B. The interaction was mediated by a region of spastin lying between residues 80 and 196 and containing a microtubule interacting and trafficking domain. Expression of epitope-tagged CHMP1B in mammalian cells prevented the development of the abnormal microtubule phenotype associated with expression of ATPase-defective spastin. These data point to a role for spastin in intracellular membrane traffic events and provide further evidence to support the emerging recognition that defects in intracellular membrane traffic are a significant cause of motor neuron pathology. PMID- 15537669 TI - Selection of optical biosensors from chemisynthetic antibody libraries. AB - We describe a method for creating antibodies with a fluorescent reporter integrated into the antigen-binding site. A reporter molecule was chemically linked to a hypervariable loop of an antibody repertoire displayed on phage, and this repertoire was selected for antigen binding. In one selected antibody, the fluorescence of the probe responded quantitatively to antigen binding. The method may have application for the engineering of homogeneous immunoassays. PMID- 15537670 TI - The 5-hydroxytryptamine4 receptor exhibits frequency-dependent properties in synaptic plasticity and behavioural metaplasticity in the hippocampal CA1 region in vivo. AB - Long-term plasticity, in the forms of long-term depression (LTD) and long-term potentiation (LTP), of synaptic transmission are thought to underlie memory. Biogenic amino acids modulate the expression of LTD and LTP. The serotonergic 5 hydroxytryptamine4 (5-HT4) receptor has been shown to influence learning and memory. However, little is known about the role of this receptor in synaptic plasticity. Here we show that although induction of LTP is unaffected by either pharmacological activation or inhibition of 5-HT4, application of the 5-HT4 receptor agonist, RS67333, completely blocks learning-induced depotentiation of LTP in the hippocampal CA1 region of freely moving rats, suggesting a role for 5 HT4 receptors in behavioural metaplasticity. In addition, the 5-HT4 antagonist RS39604 enhances the intermediate phase of LTD and converts short-term depression into persistent LTD (>24 h), suggesting a significant role for 5-HT4 receptors in the expression of LTD in CA1. Stimulation at 10 Hz causes transient synaptic depression. However, 5-HT4 antagonist application prior to 10 Hz stimulation leads to LTD, whereas agonist application leads to LTP expression. 5-HT4 receptors thus shift the frequency-response relationship for induction of plasticity. Together, these findings suggest a key role for 5-HT4 receptors in the regulation of synaptic plasticity and the determination of the particular properties of stored synaptic information. PMID- 15537671 TI - Mice lacking dopamine D2 and D3 receptors exhibit differential activation of prefrontal cortical neurons during tasks requiring attention. AB - Mice lacking dopamine D2 and D3 receptors and wildtype littermates were tested in a two-choice perceptual discrimination test that is dependent upon optimal functioning of the prefrontal cortex. Both mutants showed no deficits in attending to the perceptual stimuli or in shifting attention between stimulus dimensions. However, the performance of both mutants differed from the wildtype in different test phases. D2 mutants exhibited significant impairment in the first compound discrimination, indicating deficits in the initial acquisition of the task-governing rules. In contrast, D3 mutants performed significantly better in a set-shifting phase that required reversal learning. The higher response accuracy of D3 mutants was also accompanied by significantly increased response latency. A stereological assessment of test-induced expression of the c-fos gene in neurons of the anterior cingulate and prelimbic/infralimbic cortices revealed highest activation in D3 mutants, intermediate activation in wildtype and lowest activation in D2 mutants, indicating that response accuracy in the cognitive test correlates with the magnitude of prefrontal cortical activation regardless of which test phases revealed different performances. The study illustrates that dopamine differentially modulates prefrontal cortical activity during tasks requiring attention depending upon the type of D2-like receptor that is activated. PMID- 15537672 TI - On the benefits of not trying: brain activity and connectivity reflecting the interactions of explicit and implicit sequence learning. AB - Under certain circumstances, implicit, automatic learning may be attenuated by explicit memory processes. We explored the brain basis of this phenomenon in a functional magnetic resonance imaging (fMRI) study of motor sequence learning. Using a factorial design that crossed subjective intention to learn (explicit versus implicit) with sequence difficulty (a standard versus a more complex alternating sequence), we show that explicit attempts to learn the difficult sequence produce a failure of implicit learning and, in a follow-up behavioural experiment, that this failure represents a suppression of learning itself rather than of the expression of learning. This suppression is associated with sustained right frontal activation and attenuation of learning-related changes in the medial temporal lobe and the thalamus. Furthermore, this condition is characterized by a reversal of the fronto-thalamic connectivity observed with unimpaired implicit learning. The findings demonstrate a neural basis for a well known behavioural effect: the deleterious impact of an explicit search upon implicit learning. PMID- 15537673 TI - Focal decline of cortical thickness in Alzheimer's disease identified by computational neuroanatomy. AB - Alzheimer's disease (AD) is characterized by a heterogeneous distribution of pathological changes throughout the brain. Magnetic resonance imaging can be used to investigate the regional distribution of cortical atrophy in AD in vivo. One marker for the disease-specific atrophy is the thickness of the cortical mantle across the brain, obtained with automated 3-D image processing. Here, we present data from 36 subjects (17 controls and, 19 patients diagnosed as probable AD) investigated for cortical thickness across the entire brain. We show significant cortical thickness decline in AD in temporal, orbitofrontal and parietal regions, with the most pronounced changes occurring in the allocortical region of the medial temporal lobes, outlining the parahippocampal gyrus, and representing a loss of >1.25 millimeters of cortical thickness. Moreover, focal cortical areas decline with progression of the disease as measured by time from baseline scan as well as the Mini-Mental State Exam. The results demonstrate the ability of this method to detect changes in cortical thickness in AD, across the entire brain, without need of prior anatomical definitions. The regional distribution of changes reported here is consistent with independent findings on the distribution of neuropathological alterations in AD. Using cortical thickness, moreover, we provide a direct quantitative index of atrophy in the disease. PMID- 15537674 TI - Grabbing your ear: rapid auditory-somatosensory multisensory interactions in low level sensory cortices are not constrained by stimulus alignment. AB - Multisensory interactions are observed in species from single-cell organisms to humans. Important early work was primarily carried out in the cat superior colliculus and a set of critical parameters for their occurrence were defined. Primary among these were temporal synchrony and spatial alignment of bisensory inputs. Here, we assessed whether spatial alignment was also a critical parameter for the temporally earliest multisensory interactions that are observed in lower level sensory cortices of the human. While multisensory interactions in humans have been shown behaviorally for spatially disparate stimuli (e.g. the ventriloquist effect), it is not clear if such effects are due to early sensory level integration or later perceptual level processing. In the present study, we used psychophysical and electrophysiological indices to show that auditory somatosensory interactions in humans occur via the same early sensory mechanism both when stimuli are in and out of spatial register. Subjects more rapidly detected multisensory than unisensory events. At just 50 ms post-stimulus, neural responses to the multisensory 'whole' were greater than the summed responses from the constituent unisensory 'parts'. For all spatial configurations, this effect followed from a modulation of the strength of brain responses, rather than the activation of regions specifically responsive to multisensory pairs. Using the local auto-regressive average source estimation, we localized the initial auditory-somatosensory interactions to auditory association areas contralateral to the side of somatosensory stimulation. Thus, multisensory interactions can occur across wide peripersonal spatial separations remarkably early in sensory processing and in cortical regions traditionally considered unisensory. PMID- 15537675 TI - Structural and functional aberrations in the cerebral cortex of tenascin-C deficient mice. AB - The extracellular matrix glycoprotein tenascin-C (TNC) has been implicated in neural development and plasticity but many of its functions in vivo remain obscure. Here we addressed the question as to whether the constitutive absence of TNC in mice affects cortical physiology and structure. Defined major cell populations (neurons and inhibitory neuronal subpopulations, astrocytes, oligodendrocytes and microglia) were quantified in the somatosensory and motor cortices of adult TNC deficient (TNC-/-) and wild-type (TNC+/+) mice by immunofluorescence labelling and stereology. In both areas studied we found abnormally high neuronal density, astrogliosis, low density of parvalbumin positive interneurons and reduced ratios of oligodendrocytes to neurons and of inhibitory to excitatory neurons in the TNC deficient as opposed to the non deficient animals. Analysis of Golgi-impregnated layer V pyramidal neurons in TNC /- animals showed aberrant dendrite tortuosity and redistribution of stubby spines within first- to third-order dendritic arbors. Significantly enhanced responses upon whisker stimulation were recorded epicranially over the barrel and the motor cortices of TNC-/- as compared to TNC+/+ animals, and this effect might be associated with the diminished inhibitory circuitry. These results indicate that TNC is essential for normal cortical development and function. PMID- 15537678 TI - Thirty month outcome from early childhood head injury: a prospective analysis of neurobehavioural recovery. AB - Traumatic brain injury (TBI) is a common cause of acquired disability during childhood. While much is now known about outcome following TBI in the school-aged population, recovery in infants and young children is less well documented. The aim of this study was to examine neurobehavioural function following TBI during early childhood, to plot recovery over the 30 months post-injury and to identify predictors of outcome. The study compared three groups of children, sustaining injuries of different severity (mild = 14, moderate = 46, severe = 24), aged 2.0 6.11 years at injury, with a healthy control group (n = 33). The groups were similar with respect to pre-injury adaptive and behavioural function, psychosocial characteristics, age and gender. Using a prospective, longitudinal design, intellectual, language and memory functions were investigated acutely post-injury and again at 12 and 30 months post-injury. Results suggested a strong association between injury severity across all neurobehavioural domains. Further, 30 month outcome was predicted by multiple factors including injury severity, socio-economic status, pre-injury adaptive abilities and age, with pre-injury child behaviour and specific lesion characteristics playing surprisingly little role. In conclusion, children with more severe injuries, lower pre-injury adaptive abilities and lower socio-economic status are at greatest risk of long term neurobehavioural impairment, even several years post-injury. PMID- 15537676 TI - Ketamine disrupts frontal and hippocampal contribution to encoding and retrieval of episodic memory: an fMRI study. AB - The N-methyl-D-aspartate (NMDA) receptor antagonist ketamine produces episodic memory deficits. We used functional magnetic resonance imaging to characterize the effects of ketamine on frontal and hippocampal responses to memory encoding and retrieval in healthy volunteers using a double-blind, placebo-controlled, randomized, within-subjects comparison of two doses of intravenous ketamine. Dissociation of the effects of ketamine on encoding and retrieval processes was achieved using two study-test cycles: in the first, items were encoded prior to drug infusion and retrieval tested, during scanning, on drug; in the second, encoding was scanned on drug, and retrieval tested once ketamine plasma levels had declined. We additionally determined the interaction of ketamine with the depth of processing that occurred at encoding. A number of effects upon task dependent activations were seen. Overall, our results suggest that left frontal activation is augmented by ketamine when elaborative semantic processing is required at encoding. In addition, successful encoding on ketamine is supplemented by additional non-verbal processing that is incidental to task demands. The effects of ketamine at retrieval are consistent with impaired access to accompanying contextual features of studied items. Our findings show that, even when overt behaviour is unimpaired, ketamine has an impact upon the recruitment of key regions in episodic memory task performance. PMID- 15537679 TI - Intermittent catheterisation in older people: a valuable alternative to an indwelling catheter? AB - OBJECTIVES: To investigate whether intermittent catheterisation is a valuable alternative to an indwelling catheter in patients older than 70 years with post void residuals more than 50% of the bladder capacity. PATIENTS AND METHODS: We retrospectively reviewed the medical records of 21 patients (14 women, 7 men) older than 70 years in whom intermittent catheterisation was initiated because of voiding dysfunction with post-void residuals more than 50% of the bladder capacity resistant to other treatment. Twelve patients mastered the technique of intermittent self-catheterisation, seven were catheterised by their partners and two by nurses. RESULTS: The mean age of patients was 76.5 years (range 71-83 years) and the mean observation period with regard to intermittent catheterisation was 27.9 months (range 5-129 months). For those relying on intermittent catheterisation, the urinary tract infection rate was 0.84 per year and patient (range 0-3), and urinary continence was restored in all of the six previously incontinent patients. Eighteen of the 21 patients reported a significantly improved quality of life owing to the restoration of urinary continence, decreasing of daytime frequency, nocturia and urge, and the lowering of the urinary tract infection rate. CONCLUSIONS: Intermittent (self-) catheterisation is a safe and valuable technique in older people with significant post-void residuals owing to detrusor underactivity. Urinary continence is restored, urge, daytime frequency and nocturia are decreased, and the urinary tract infection rate is diminished, resulting in improved quality of life. Therefore, intermittent (self-) catheterisation is strongly recommended in older people. PMID- 15537680 TI - Occupational therapy compared with social work assessment for older people. An economic evaluation alongside the CAMELOT randomised controlled trial. AB - OBJECTIVES: To compare costs and outcome of occupational therapy-led assessment with social worker-led assessment of older people, in terms of their independence and quality of life. DESIGN: Cost-effectiveness analysis alongside a randomised controlled trial. The analysis took viewpoints of health services and patients. The primary outcome measure for cost-effectiveness was dependency using the Community Dependency Index (CDI). Secondary outcomes included utility scores based on the EuroQoL (EQ-5D). Resource use was measured for each patient, from clinical records and from patient carer interviews at 8 months. Unit costs of health and social care resources were derived from local sources and national datasets. Cost-effectiveness was analysed using cost-effectiveness acceptability curves. RESULTS: There were no differences between the two arms of the trial in terms of cost-effectiveness. There is an apparent increase in mean cost per case for the occupational therapy arm but this is not statistically significant (mean difference in cost per case 542 pounds, 95% CI 434-1,519 pounds). Mean total costs of care per participant were 4,379 pounds and 3,837 pounds for the occupational therapy and social work arms, respectively. At best the intervention would improve outcomes at a cost of 14,000 pounds per quality-adjusted life year (QALY). The probability of such an outcome was <50%. CONCLUSIONS: From a policy perspective, the lack of difference in clinical and cost-effectiveness means that either a social work or an occupational therapy service is successful in making care assessments that enable an older person to remain in their own home. PMID- 15537681 TI - Arterial Biology for the Investigation of the Treatment Effects of Reducing Cholesterol (ARBITER) 2: a double-blind, placebo-controlled study of extended release niacin on atherosclerosis progression in secondary prevention patients treated with statins. AB - BACKGROUND: Niacin reduces coronary heart disease morbidity and mortality when taken either alone or in combination with statins; however, the incremental impact of adding niacin to background statin therapy is unknown. METHODS AND RESULTS: This was a double-blind randomized placebo-controlled study of once daily extended-release niacin (1000 mg) added to background statin therapy in 167 patients (mean age 67 years) with known coronary heart disease and low levels of high-density lipoprotein cholesterol (HDL-C; <45 mg/dL). The primary end point was the change in common carotid intima-media thickness (CIMT) after 1 year. Baseline CIMT (0.884+/-0.234 mm), low-density lipoprotein cholesterol (89+/-20 mg/dL), and HDL-C (40+/-7 mg/dL) were comparable in the placebo and niacin groups. Adherence to niacin exceeded 90%, and 149 patients (89.2%) completed the study. HDL-C increased 21% (39 to 47 mg/dL) in the niacin group. After 12 months, mean CIMT increased significantly in the placebo group (0.044+/-0.100 mm; P<0.001) and was unchanged in the niacin group (0.014+/-0.104 mm; P=0.23). Although the overall difference in IMT progression between the niacin and placebo groups was not statistically significant (P=0.08), niacin significantly reduced the rate of IMT progression in subjects without insulin resistance (P=0.026). Clinical cardiovascular events occurred in 3 patients treated with niacin (3.8%) and 7 patients treated with placebo (9.6%; P=0.20). CONCLUSIONS: The addition of extended-release niacin to statin therapy slowed the progression of atherosclerosis among individuals with known coronary heart disease and moderately low HDL-C. PMID- 15537682 TI - Meta-analysis: high-dosage vitamin E supplementation may increase all-cause mortality. AB - BACKGROUND: Experimental models and observational studies suggest that vitamin E supplementation may prevent cardiovascular disease and cancer. However, several trials of high-dosage vitamin E supplementation showed non-statistically significant increases in total mortality. PURPOSE: To perform a meta-analysis of the dose-response relationship between vitamin E supplementation and total mortality by using data from randomized, controlled trials. PATIENTS: 135,967 participants in 19 clinical trials. Of these trials, 9 tested vitamin E alone and 10 tested vitamin E combined with other vitamins or minerals. The dosages of vitamin E ranged from 16.5 to 2000 IU/d (median, 400 IU/d). DATA SOURCES: PubMed search from 1966 through August 2004, complemented by a search of the Cochrane Clinical Trials Database and review of citations of published reviews and meta analyses. No language restrictions were applied. DATA EXTRACTION: 3 investigators independently abstracted study reports. The investigators of the original publications were contacted if required information was not available. DATA SYNTHESIS: 9 of 11 trials testing high-dosage vitamin E (> or =400 IU/d) showed increased risk (risk difference > 0) for all-cause mortality in comparisons of vitamin E versus control. The pooled all-cause mortality risk difference in high dosage vitamin E trials was 39 per 10,000 persons (95% CI, 3 to 74 per 10,000 persons; P = 0.035). For low-dosage vitamin E trials, the risk difference was -16 per 10,000 persons (CI, -41 to 10 per 10,000 persons; P > 0.2). A dose-response analysis showed a statistically significant relationship between vitamin E dosage and all-cause mortality, with increased risk of dosages greater than 150 IU/d. LIMITATIONS: High-dosage (> or =400 IU/d) trials were often small and were performed in patients with chronic diseases. The generalizability of the findings to healthy adults is uncertain. Precise estimation of the threshold at which risk increases is difficult. CONCLUSION: High-dosage (> or =400 IU/d) vitamin E supplements may increase all-cause mortality and should be avoided. PMID- 15537683 TI - Summaries for patients. Vitamin E supplements may be harmful. PMID- 15537684 TI - Repression of Dpp targets in the Drosophila wing by Brinker. AB - Patterning along developing body axes is regulated by gradients of transcription factors, which activate or repress different genes above distinct thresholds. Understanding differential threshold responses requires knowledge of how these factors regulate transcription. In the Drosophila wing, expression of genes such as omb and sal along the anteroposterior axis is restricted by lateral-to-medial gradients of the transcriptional repressor Brinker (Brk). omb is less sensitive to repression by Brk than sal and is consequently expressed more laterally. Contrary to previous suggestions, we show that Brk cannot repress simply by competing with activators, but requires specific repression domains along with its DNA-binding domain. Brk possesses at least three repression domains, but these are not equivalent; one, 3R, is sufficient to repress omb but not sal. Thus, although sal and omb show quantitative differences in their response to Brk, there are qualitative differences in the mechanisms that Brk uses to repress them. PMID- 15537685 TI - alpha3beta1 integrin modulates neuronal migration and placement during early stages of cerebral cortical development. AB - We show that alpha3 integrin mutation disrupts distinct aspects of neuronal migration and placement in the cerebral cortex. The preplate develops normally in alpha3 integrin mutant mice. However, time lapse imaging of migrating neurons in embryonic cortical slices indicates retarded radial and tangential migration of neurons, but not ventricular zone-directed migration. Examination of the actin cytoskeleton of alpha3 integrin mutant cortical cells reveals aberrant actin cytoskeletal dynamics at the leading edges. Deficits are also evident in the ability of developing neurons to probe their cellular environment with filopodial and lamellipodial activity. Calbindin or calretinin positive upper layer neurons as well as the deep layer neurons of alpha3 integrin mutant mice expressing EGFP were misplaced. These results suggest that alpha3beta1 integrin deficiency impairs distinct patterns of neuronal migration and placement through dysregulated actin dynamics and defective ability to search and respond to migration modulating cues in the developing cortex. PMID- 15537686 TI - The Drosophila MBD2/3 protein mediates interactions between the MI-2 chromatin complex and CpT/A-methylated DNA. AB - Methyl-DNA binding proteins play an important role in epigenetic gene regulation. The Drosophila genome encodes a single protein (MBD2/3) with extended homologies to the vertebrate methyl-DNA binding proteins MBD2 and MBD3. However, very little is known about its functional properties. We have now characterized an MBD2/3 null mutant allele that is viable and fertile. This mutation caused a strong dominant suppression of position-effect variegation and also resulted in a high rate of chromosome segregation defects during early embryogenesis. Confocal analysis of mutant embryos showed local displacement of MI-2 from DNA and indicated that MBD2/3 is associated with only a subset of MI-2 complexes. In addition, band shift experiments demonstrated a specific binding of MBD2/3 to CpT/A-methylated DNA, which reflects the endogenous DNA methylation pattern of Drosophila. Consistently, the localization of MBD2/3 was disrupted in embryos with reduced levels of DNA methylation. Our data provide novel insights into the function of MBD2/3 proteins and strongly suggest the existence of methylation dependent chromatin structures in Drosophila. PMID- 15537687 TI - Coordinated development of muscles and tendons of the Drosophila leg. AB - Since Miller's morphological description, the Drosophila leg musculature and its formation has not been revisited. Here, using a set of GFP markers and confocal microscopy, we analyse Drosophila leg muscle development, and describe all the muscles and tendons present in the adult leg. Importantly, we provide for the first time evidence for tendons located internally within leg segments. By visualising muscle and tendon precursors, we demonstrate that leg muscle development is closely associated with the formation of internal tendons. In the third instars discs, in the vicinity of tendon progenitors, some Twist-positive myoblasts start to express the muscle founder cell marker dumbfounded (duf). Slightly later, in the early pupa, epithelial tendon precursors invaginate inside the developing leg segments, giving rise to the internal string-like tendons. The tendon-associated duf-lacZ-expressing muscle founders are distributed along the invaginating tendon precursors and then fuse with surrounding myoblasts to form syncytial myotubes. At mid-pupation, these myotubes grow towards their epithelial insertion sites, apodemes, and form links between internally located tendons and the leg epithelium. This leads to a stereotyped pattern of multifibre muscles that ensures movement of the adult leg. PMID- 15537688 TI - Pigment pattern evolution by differential deployment of neural crest and post embryonic melanophore lineages in Danio fishes. AB - Latent precursors or stem cells of neural crest origin are present in a variety of post-embryonic tissues. Although these cells are of biomedical interest for roles in human health and disease, their potential evolutionary significance has been underappreciated. As a first step towards elucidating the contributions of such cells to the evolution of vertebrate form, we investigated the relative roles of neural crest cells and post-embryonic latent precursors during the evolutionary diversification of adult pigment patterns in Danio fishes. These pigment patterns result from the numbers and arrangements of embryonic melanophores that are derived from embryonic neural crest cells, as well as from post-embryonic metamorphic melanophores that are derived from latent precursors of presumptive neural crest origin. In the zebrafish D. rerio, a pattern of melanophore stripes arises during the larval-to-adult transformation by the recruitment of metamorphic melanophores from latent precursors. Using a comparative approach in the context of new phylogenetic data, we show that adult pigment patterns in five additional species also arise from metamorphic melanophores, identifying this as an ancestral mode of adult pigment pattern development. By contrast, superficially similar adult stripes of D. nigrofasciatus (a sister species to D. rerio) arise by the reorganization of melanophores that differentiated at embryonic stages, with a diminished contribution from metamorphic melanophores. Genetic mosaic and molecular marker analyses reveal evolutionary changes that are extrinsic to D. nigrofasciatus melanophore lineages, including a dramatic reduction of metamorphic melanophore precursors. Finally, interspecific complementation tests identify a candidate genetic pathway for contributing to the evolutionary reduction in metamorphic melanophores and the increased contribution of early larval melanophores to D. nigrofasciatus adult pigment pattern development. These results demonstrate an important role for latent precursors in the diversification of pigment patterns across danios. More generally, differences in the deployment of post-embryonic neural crest-derived stem cells or their specified progeny may contribute substantially to the evolutionary diversification of adult form in vertebrates, particularly in species that undergo a metamorphosis. PMID- 15537689 TI - Conservation of B-class floral homeotic gene function between maize and Arabidopsis. AB - The ABC model of flower development, established through studies in eudicot model species, proposes that petal and stamen identity are under the control of B-class genes. Analysis of B- and C-class genes in the grass species rice and maize suggests that the C- and B-class functions are conserved between monocots and eudicots, with B-class genes controlling stamen and lodicule development. We have undertaken a further analysis of the maize B-class genes Silky1, the putative AP3 ortholog, and Zmm16, a putative PI ortholog, in order to compare their function with the Arabidopsis B-class genes. Our results show that maize B-class proteins interact in vitro to bind DNA as an obligate heterodimer, as do Arabidopsis B class proteins. The maize proteins also interact with the appropriate Arabidopsis B-class partner proteins to bind DNA. Furthermore, we show that maize B-class genes are capable of rescuing the corresponding Arabidopsis B-class mutant phenotypes. This demonstrates B-class activity of the maize gene Zmm16, and provides compelling evidence that B-class gene function is conserved between monocots and eudicots. PMID- 15537690 TI - Segment-specific prevention of pioneer neuron apoptosis by cell-autonomous, postmitotic Hox gene activity. AB - In vertebrates, neurons often undergo apoptosis after differentiating and extending their axons. By contrast, in the developing nervous system of invertebrate embryos apoptosis typically occurs soon after cells are generated. Here, we show that the Drosophila dMP2 and MP1 pioneer neurons undergo segment specific apoptosis at late embryonic stages, long after they have extended their axons and have performed their pioneering role in guiding follower axons. This segmental specificity is achieved by differential expression of the Hox gene Abdominal B, which in posterior segments prevents pioneer neuron death postmitotically and cell-autonomously by repressing the RHG-motif cell death activators reaper and grim. Our results identify the first clear case of a cell autonomous and anti-apoptotic role for a Hox gene in vivo. In addition, they provide a novel mechanism linking Hox positional information to differences in neuronal architecture along the anteroposterior axis by the selective elimination of mature neurons. PMID- 15537691 TI - IMP-1 metallo-beta-lactamase-producing Pseudomonas aeruginosa in a university hospital in the People's Republic of China. PMID- 15537692 TI - Increase and change in pattern of hospital antimicrobial use, Denmark, 1997-2001. AB - OBJECTIVES: To analyse the changes and compare antimicrobial consumption in public hospitals in Denmark over the period 1997-2001. METHODS: Data on the number of WHO defined daily doses (DDD) were obtained from the Danish Medicines Agency. Data on the number of bed-days were obtained from the National Board of Health. We calculated antimicrobial consumption in hospitals as the number of DDD per 100 bed-days for all antibacterials for systemic use i.e. group J01 of the Anatomical Therapeutic Chemical (ATC) classification and for classes of this group. RESULTS: During 1997-2001, antimicrobial use in hospitals in Denmark significantly increased by 18%, from 38.0 to 44.8 DDD per 100 bed-days (P < 0.005). Most of this increase (55%) was attributed to an increase in consumption of commonly used classes of antimicrobials, mainly penicillins with extended spectrum (ATC group J01CA), beta-lactamase-sensitive penicillins (J01CE) and beta lactamase-resistant penicillins (J01CF). The 'broad-spectrum' and newer antimicrobials, i.e. combinations of penicillins with beta-lactamase inhibitor (J01CR), cephalosporins (J01DA), carbapenems (J01DH) and fluoroquinolones (J01MA) contributed to 36% of the increase. Together, these amounted to 16% of total consumption in hospitals in Denmark in 1997, rising to 19% in 2001. CONCLUSIONS: Although antimicrobial consumption in public hospitals in Denmark is low compared with other countries, the steady increase and change in pattern of their use are causes of concern, deserving close monitoring and further investigations. PMID- 15537693 TI - A new phenotype of resistance to lincosamide and streptogramin A-type antibiotics in Streptococcus agalactiae in New Zealand. AB - OBJECTIVES: To characterize a new type of resistance to clindamycin in Streptococcus agalactiae. METHODS: Nineteen erythromycin-susceptible, clindamycin resistant S. agalactiae isolates from New Zealand were studied. MICs of macrolide, lincosamide and streptogramin antibiotics were determined. Clindamycin and streptogramin resistance genes were searched for by PCR. Isolates were compared by serotyping and by DNA macrorestriction patterns determined by PFGE. Conjugative transfer of resistance traits to recipient strains of S. agalactiae and Enterococcus faecium was assayed. RESULTS: The 19 S. agalactiae isolates were intermediate or resistant to clindamycin (MIC range: 0.5-2 mg/L) and lincomycin (MIC range: 1-8 mg/L) and had high MICs of dalfopristin (4-32 mg/L), a streptogramin A-type antibiotic, compared with controls. By contrast, the strains were susceptible to macrolides and quinupristin, a streptogramin B-type antibiotic. This new phenotype was called LSA (lincosamide-streptogramin A). Clindamycin resistance could not be transferred to recipient strains. Thirteen isolates belonged to serotype III and to a single PFGE genotype A, and five isolates belonged to serotype I and to genotype B. One isolate was non-typeable and belonged to a distinct genotype C. CONCLUSIONS: We have characterized a new LSA phenotype in S. agalactiae. Analysis of restriction patterns of S. agalactiae chromosomal DNA showed that the resistance was spread in a minimum of three bacterial clones. The genetic and biochemical basis for the resistance remains unknown. PMID- 15537694 TI - Plasma levels of zidovudine twice daily compared with three times daily in six HIV-1-infected children. AB - OBJECTIVES: Zidovudine is often administered every 12 h in HIV-infected children, but so far no pharmacokinetic data are available for the administration of this agent every 12 h. We have evaluated the plasma pharmacokinetics of zidovudine administered every 8 h versus every 12 h in HIV-1-infected children. METHODS: In HIV-1-infected children who switched from zidovudine every 8 h to every 12 h, a pharmacokinetic curve was recorded both before and after the switch. Zidovudine plasma levels were measured by HPLC. Pharmacokinetic parameters were calculated by non-compartmental methods. RESULTS: Six HIV-1-infected children [median age (range) 7.8 (2.5-13.4) years] were included. In these patients, geometric mean ratios of AUC(0-24) and C(max) for zidovudine every 12 h versus every 8 h were not significantly different from 1.0. CONCLUSIONS: The plasma pharmacokinetic parameters of zidovudine taken every 8 h and every 12 h were not significantly different and therefore suggest bioequivalence of these two dose frequencies. PMID- 15537695 TI - The intracellular pharmacology of antiretroviral protease inhibitors. AB - Therapeutic drug monitoring (TDM) of antiretroviral protease inhibitors (PIs) has been suggested to have the potential to both reduce toxicity and optimize individual therapy. However, the major target of PIs is within cells infected with HIV. Therefore clinical outcome ultimately must be related to intracellular drug concentrations since antiviral activity of PIs is highly correlated with intracellular concentrations in vitro. Intracellular pharmacokinetics provides information regarding drug disposition in a compartment where HIV replication occurs and combined with plasma data may be useful in understanding therapeutic failure in relation to cellular resistance. In order to improve therapeutic efficacy, it is therefore important that the intracellular pharmacokinetics of drugs, such as PIs, is studied in addition to plasma pharmacokinetics. Multidrug resistance transporters may result in a lower cellular concentration of drug via an efflux mechanism, thus contributing to sanctuary site formation. However, conclusive proof that transporters contribute to clinical drug resistance is still lacking, although recent studies have attempted to address this issue. In relation to host and cellular factors, this review considers several issues involved in influencing intracellular drug concentrations and discusses the intracellular levels of PIs recently published from cellular studies. PMID- 15537696 TI - Reduced colonization and infection with miconazole-rifampicin modified central venous catheters: a randomized controlled clinical trial. AB - OBJECTIVE: Central venous catheters (CVC) are a major cause of nosocomial bloodstream infections. Catheters modified with miconazole and rifampicin that constantly and slowly release antimicrobial substances are assumed to be beneficial in reducing rates of colonization and catheter-related infections. DESIGN AND SETTING: Prospective controlled non-blinded randomized clinical trial in two German university hospitals. PATIENTS: 223 adult inpatients with CVC between October 2000 and February 2002. Baseline characteristics, APACHE II score and therapeutic interventions were comparable. INTERVENTION: Randomization to receive either a miconazole and rifampicin modified catheter (n=118) or a standard triple-lumen CVC (n=105). MEASUREMENTS, DEFINITIONS: Microbiological evaluation was done after CVC removal. A catheter was considered colonized if growth of > or =15 cfu was found by semi-quantitative roll-plate technique from a proximal or distal catheter segment. A catheter-related infection (CRI) was defined as a colonized catheter with local signs of inflammation. A catheter related bloodstream infection (CR-BSI) was defined as a colonized catheter with isolation of the same organism from the patient's blood with accompanying clinical signs of infection. RESULTS: A colonization of CVC was observed in six patients (5.1%) with a modified catheter and 38 patients (36.2%) with a standard catheter (P < 0.001). Five patients in the modified group (4.2%) and 18 in the standard group (17.1%) developed CRI (P=0.002). One assumed CR-BSI was detected in the standard group, with none in the modified group. No adverse effects related to the modified catheters and no antimicrobial resistance were observed. CONCLUSION: CVC supersaturated with miconazole and rifampicin were associated with a significantly lower risk for catheter colonization and catheter-related infections compared to standard catheters. PMID- 15537697 TI - Silver nanoparticles and polymeric medical devices: a new approach to prevention of infection? AB - OBJECTIVES: Implantable devices are major risk factors for hospital-acquired infection. Biomaterials coated with silver oxide or silver alloy have all been used in attempts to reduce infection, in most cases with controversial or disappointing clinical results. We have developed a completely new approach using supercritical carbon dioxide to impregnate silicone with nanoparticulate silver metal. This study aimed to evaluate the impregnated polymer for antimicrobial activity. METHODS: After impregnation the nature of the impregnation was determined by transmission electron microscopy. Two series of polymer discs were then tested, one washed in deionized water and the other unwashed. In each series, half of the discs were coated with a plasma protein conditioning film. The serial plate transfer test was used as a screen for persisting activity. Bacterial adherence to the polymers and the rate of kill, and effect on planktonic bacteria were measured by chemiluminescence and viable counts. Release rates of silver ions from the polymers in the presence and absence of plasma was measured using inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Tests for antimicrobial activity under various conditions showed mixed results, explained by the modes and rates of release of silver ions. While washing removed much of the initial activity there was continued release of silver ions. Unexpectedly, this was not blocked by conditioning film. CONCLUSIONS: The methodology allows for the first time silver impregnation (as opposed to coating) of medical polymers and promises to lead to an antimicrobial biomaterial whose activity is not restricted by increasing antibiotic resistance. PMID- 15537698 TI - Antimicrobial efficacy of a new antibiotic-loaded poly(hydroxybutyric-co hydroxyvaleric acid) controlled release system. AB - OBJECTIVE: Failure of orthopaedic devices, mainly femoral hip replacements, due to infection is of increasing medical importance. There is a need for improved antibiotic delivery systems in the treatment of orthopaedic infections and here we have evaluated polyhydroxyalkanoate formulations for their suitability as a constant delivery system for gentamicin. METHODS: Gentamicin was incorporated in poly(hydroxybutyric-co-hydroxyvalerate) (PHBV) with 8% or 12% hydroxyvalerate (HV) content at 2:1 or 5:1 (weight to weight) ratio. In conjunction with an elution study, a scanning electron microscopy and a porosity study were carried out to explore physical characteristics of the complexes before and after the leaching effect. The antibacterial effectiveness of the complexes was analysed in a bacterial adhesion assay using clinical isolates of Staphylococcus haemolyticus and Staphylococcus aureus. In addition, the polymers were exposed to pooled human blood to test their biocompatibility in both static and dynamic environments. RESULTS: We have shown that increasing the HV content from 8% to 12% leads to a faster release of the integrated antibiotic. An increase in antibiotic content enhanced the homogeneity while decreasing the permeability of the complexes and reducing the release rate. A significant reduction in the number of the adherent S. aureus and gentamicin-resistant S. haemolyticus within a 48 h exposure to our formulations confirmed the effectiveness of the PHBV/gentamicin complexes. Finally, these formulations did not alter the haemodynamics of the pooled blood samples after an extended period of time. CONCLUSION: Taken together, the PHBV/gentamicin formulations may prove to be effective preventive therapeutic modalities in implant-related Staphylococcus infections. PMID- 15537699 TI - Antimicrobial resistance in Campylobacter strains isolated from French broilers before and after antimicrobial growth promoter bans. AB - OBJECTIVES: The antimicrobial susceptibility of Campylobacter strains isolated from standard and free-range broilers in 1992-1996 and 2001-2002 was studied. METHODS: Strains were isolated from caeca or skin samples collected from standard or free-range broilers arriving in slaughterhouses. The MICs of ampicillin, nalidixic acid, enrofloxacin, tetracycline, erythromycin and gentamicin were determined by agar dilution and compared according to species (Campylobacter jejuni or Campylobacter coli), production system and sampling period. RESULTS: Results showed that all chickens harboured Campylobacter. An increase over time of the C. coli/C. jejuni ratio for standard chickens occurred. A wide range of MICs was observed among isolates from the same broiler or from the same farm. Strains collected on entry to the slaughterhouse and after storage showed no significant difference in their antibiotic resistance. C. coli was more resistant than C. jejuni to tetracycline and erythromycin during the first period and to all tested molecules (except gentamicin) during the second period. Strains isolated from standard chickens were also more often resistant than those isolated from free-range broilers. The percentage of C. jejuni strains resistant to ampicillin decreased from 1992-1996 to 2001-2002, whereas no change could be observed for the other antimicrobial agents. However, for C. coli the resistance to ampicillin, nalidixic acid, enrofloxacin, tetracycline and erythromycin significantly increased. CONCLUSION: There was an increase in the incidence of antibiotic resistance of C. coli between 1992-1996 and 2001-2002. PMID- 15537700 TI - In vivo comparison of Dhvar-5 and gentamicin in an MRSA osteomyelitis prevention model. AB - OBJECTIVES: The continued rise in drug-resistant pathogens has led to global research efforts into new antimicrobial agents. A promising class of new agents are the antimicrobial peptides. The aim of the study was to investigate the efficacy of the antimicrobial peptide Dhvar-5 in a prophylactic, methicillin resistant Staphylococcus aureus (MRSA) osteomyelitis model. METHODS: Dhvar-5 (12 mg or 24 mg/rabbit) was incorporated into polymethyl methacrylate (PMMA) beads as a local drug delivery system. For comparison, plain beads (control) and beads containing gentamicin as a sulphate (10 mg or 24 mg per rabbit) were also prepared. The beads were inserted into the inoculated femoral cavity of 36 rabbits, and 1 week later they were killed. The presence and severity of MRSA osteomyelitis was assessed by culture and histology. RESULTS: Both the 24 mg Dhvar-5 beads and the 24 mg gentamicin sulphate beads significantly reduced the bacterial load of the inoculated femora compared with the control chain. Although a 24 mg Dhvar-5 dose inhibited MRSA growth, it did not completely sterilize the femora. Sterilization occurred only in some of the gentamicin-treated specimens. CONCLUSION: We conclude that both the gentamicin beads and the Dhvar-5 beads were only partially effective at preventing MRSA infection in this model. PMID- 15537701 TI - Genetic analysis of the neuronal and ubiquitous AP-3 adaptor complexes reveals divergent functions in brain. AB - Neurons express adaptor (AP)-3 complexes assembled with either ubiquitous (beta3A) or neuronal-specific (beta3B) beta3 isoforms. However, it is unknown whether these complexes indeed perform distinct functions in neuronal tissue. Here, we explore this hypothesis by using genetically engineered mouse models lacking either beta3A- or beta3B-containing AP-3 complexes. Somatic and neurological phenotypes were specifically associated with the ubiquitous and neuronal adaptor deficiencies, respectively. At the cellular level, AP-3 isoforms were localized to distinct neuronal domains. beta3B-containing AP-3 complexes were preferentially targeted to neuronal processes. Consistently, beta3B deficiency compromised synaptic zinc stores assessed by Timm's staining and the synaptic vesicle targeting of membrane proteins involved in zinc uptake (ZnT3 and ClC-3). Surprisingly, despite the lack of neurological symptoms, beta3A-deficient mouse brain possessed significantly increased synaptic zinc stores and synaptic vesicle content of ZnT3 and ClC-3. These observations indicate that the functions of beta3A- and beta3B-containing complexes are distinct and divergent. Our results suggest that concerted nonredundant functions of neuronal and ubiquitous AP-3 provide a mechanism to control the levels of selected membrane proteins in synaptic vesicles. PMID- 15537702 TI - Loss of rereplication control in Saccharomyces cerevisiae results in extensive DNA damage. AB - To maintain genome stability, the entire genome of a eukaryotic cell must be replicated once and only once per cell cycle. In many organisms, multiple overlapping mechanisms block rereplication, but the consequences of deregulating these mechanisms are poorly understood. Here, we show that disrupting these controls in the budding yeast Saccharomyces cerevisiae rapidly blocks cell proliferation. Rereplicating cells activate the classical DNA damage-induced checkpoint response, which depends on the BRCA1 C-terminus checkpoint protein Rad9. In contrast, Mrc1, a checkpoint protein required for recognition of replication stress, does not play a role in the response to rereplication. Strikingly, rereplicating cells accumulate subchromosomal DNA breakage products. These rapid and severe consequences suggest that even limited and sporadic rereplication could threaten the genome with significant damage. Hence, even subtle disruptions in the cell cycle regulation of DNA replication may predispose cells to the genomic instability associated with tumorigenesis. PMID- 15537703 TI - The nuclear kinase Lsk1p positively regulates the septation initiation network and promotes the successful completion of cytokinesis in response to perturbation of the actomyosin ring in Schizosaccharomyces pombe. AB - Cytokinesis in fission yeast requires the function of an actomyosin-based contractile ring whose constriction is dependent on a signaling module termed the septation initiation network (SIN). In response to minor perturbation of the ring, the duration of SIN signaling is extended concurrently with a delay in nuclear cycle progression. These mechanisms require the conserved phosphatase Clp1p/Flp1p and facilitate the successful completion of cytokinesis, thereby increasing cellular viability. To isolate novel components of this cytokinesis monitoring system, we screened a genome-wide bank of protein kinase deletion mutants and identified Lsk1p, a nuclear-localized protein kinase. Similar to clp1Delta mutants, and in contrast to wild type, lsk1Delta cells are unable to maintain the integrity of the actomyosin ring upon treatment with low doses (0.3 microM) of latrunculin A. However, unlike clp1Delta mutants, lsk1Delta cells are competent to delay nuclear cycle progression after cytokinetic failure. In addition, lsk1Delta mutants suppress the lethal, multiseptate phenotype conferred by hyperactivation of the SIN, demonstrating that Lsk1p is a positive regulator of this module. In this report, we demonstrate that Lsk1p acts in parallel to Clp1p to promote actomyosin ring stability upon checkpoint activation. Our studies also establish that actomyosin ring maintenance and nuclear cycle delay in response to cytokinetic perturbation can be genetically resolved into independent pathways. PMID- 15537704 TI - The major target of the endogenously generated reactive oxygen species in response to insulin stimulation is phosphatase and tensin homolog and not phosphoinositide-3 kinase (PI-3 kinase) in the PI-3 kinase/Akt pathway. AB - Phosphoinositide-3 kinase (PI-3 kinase) and its downstream signaling molecules PDK-1 and Akt were analyzed in SK-N-SH and SK-N-BE(2) human neuroblastoma cell lines. When cells were stimulated with insulin, PI-3 kinase was activated in both cell lines, whereas the translocation of PDK-1 to the membrane fraction and phosphorylated Akt were observed only in SK-N-SH cells. Analyses of the insulin mediated reactive oxygen species (ROS) generation and Phosphatase and Tensin homolog (PTEN) oxidation indicate that PTEN oxidation occurred in SK-N-SH cells, which can produce ROS, but not in SK-N-BE(2) cells, which cannot increase ROS in response to insulin stimulation. When SK-N-SH cells were pretreated with the NADPH oxidase inhibitor diphenyleneiodonium chloride before insulin stimulation, insulin-mediated translocation of PDK-1 to the membrane fraction and phosphorylation of Akt were remarkably reduced, whereas PI-3 kinase activity was not changed significantly. These results indicate that not only PI-3 kinase activation but also inhibition of PTEN by ROS is needed to increase cellular level of phosphatidylinositol 3,4,5-trisphosphate for recruiting downstream signaling molecules such as PDK-1 and Akt in insulin-mediated signaling. Moreover, the ROS generated by insulin stimulation mainly contributes to the inactivation of PTEN and not to the activation of PI-3 kinase in the PI-3 kinase/Akt pathway. PMID- 15537705 TI - Ypt31/32 GTPases and their novel F-box effector protein Rcy1 regulate protein recycling. AB - Ypt/Rab GTPases control various aspects of vesicle formation and targeting via their diverse effectors. We report a new role for these GTPases in protein recycling through a novel effector. The F-box protein Rcy1, which mediates plasma membrane recycling, is identified here as a downstream effector of the Ypt31/32 GTPase pair because it binds active GTP-bound Ypt31/32 and colocalizes with these GTPases on late Golgi and endosomes. Furthermore, Ypt31/32 regulates the polarized localization and half-life of Rcy1. This suggests that Ypt/Rabs can regulate the protein level of their effectors, in addition to the established ways by which they control their effectors. We show that like Rcy1, Ypt31/32 regulate the coupled phosphorylation and recycling of the plasma membrane v-SNARE Snc1. Moreover, Ypt31/32 and Rcy1 regulate the recycling of the furin-homolog Kex2 to the Golgi. Therefore, Ypt31/32 and Rcy1 mediate endosome-to-Golgi transport, because this is the only step shared by Snc1 and Kex2. Finally, we show that Rcy1 physically interacts with Snc1. Based on this result and because F box proteins serve as adaptors between specific substrates and ubiquitin ligases, we propose that Ypt31/32 GTPases regulate the function of Rcy1 in the phosphorylation and/or ubiquitination of proteins that recycle through the Golgi. PMID- 15537706 TI - Regulation of intracellular Ca2+ release in corpus cavernosum smooth muscle: synergism between nitric oxide and cGMP. AB - Tonic contraction of corpus cavernosum smooth muscle cells (SMCs) maintains the flaccid state of the penis, and relaxation is initiated by nitric oxide (NO), leading to erection. Our aim was to investigate the effect of NO on the smooth muscle cellular response to adrenergic stimulation in corpus cavernosum. Fura-2 fluorescence was used to record intracellular Ca(2+) concentration ([Ca(2+)](i)) from freshly isolated SMCs from rat and human. Phenylephrine (PE) transiently elevated [Ca(2+)](i) in the presence and absence of extracellular Ca(2+), indicating release from intracellular stores. Whereas the NO donor S-nitroso-N acetylpenicillamine (SNAP) with sildenafil citrate (SIL) caused no change in basal [Ca(2+)](i), the PE-induced rise of [Ca(2+)](i) was reversibly inhibited by 27 +/- 7% (n = 21, P < 0.005) in rat and by 55 +/- 15% (n = 9, P < 0.01) in human SMCs. SNAP and SIL also reduced the contractile response to PE. To investigate the mechanism, we applied mediators alone or in combination. The soluble guanylyl cyclase inhibitor ODQ reduced the effect of SNAP and SIL. SIL, cGMP analogs, and NO donors without SIL did not reduce the PE-induced rise of [Ca(2+)](i). However, the combination of 8-bromo-cGMP with SNAP reduced the Ca(2+) peak by 42 +/- 9% (n = 22, P < 0.01). Our results demonstrate that NO and cGMP act synergistically to reduce Ca(2+) release from intracellular stores. Reduction of intracellular Ca(2+) release may contribute to relaxation of the corpus cavernosum, leading to erection. PMID- 15537707 TI - Kinetics, molecular basis, and differentiation of L-lactate transport in spermatogenic cells. AB - Round spermatid energy metabolism is closely dependent on the presence of L lactate in the external medium. This L-lactate has been proposed to be supplied by Sertoli cells in the seminiferous tubules. L-Lactate, in conjunction with glucose, modulates intracellular Ca(2+) concentration in round spermatids and pachytene spermatocytes. In spite of this central role of L-lactate in spermatogenic cell physiology, the mechanism of L-lactate transport, as well as possible differentiation during spermatogenesis, has not been studied in these cells. By measuring radioactive L-lactate transport and intracellular pH (pH(i)) changes with pH(i) fluorescent probes, we show that these cells transport L lactate using monocarboxylate-H(+) transport (MCT) systems. RT-PCR, in situ mRNA hybridization, and immunocyto- and immunohistochemistry data show that pachytene spermatocytes express mainly the MCT1 and MCT4 isoforms of the transporter (intermediate- and low-affinity transporters, respectively), while round spermatids, besides MCT1 and MCT4, also show expression of the MCT2 isoform (high affinity transporter). These molecular data are consistent with the kinetic data of L-lactate transport in these cells demonstrating at least two transport components for L-lactate. These separate transport components reflect the ability of these cells to switch between the generation of glycolytic L-lactate in the presence of external glucose and the use of L-lactate when this substrate is available in the external environment. The supply of these substrates is regulated by the hormonal control of Sertoli cell glycolytic activity. PMID- 15537708 TI - Pacemaker potentials generated by interstitial cells of Cajal in the murine intestine. AB - Pacemaker potentials were recorded in situ from myenteric interstitial cells of Cajal (ICC-MY) in the murine small intestine. The nature of the two components of pacemaker potentials (upstroke and plateau) were investigated and compared with slow waves recorded from circular muscle cells. Pacemaker potentials and slow waves were not blocked by nifedipine (3 microM). In the presence of nifedipine, mibefradil, a voltage-dependent Ca(2+) channel blocker, reduced the amplitude, frequency, and rate of rise of upstroke depolarization (dV/dt(max)) of pacemaker potentials and slow waves in a dose-dependent manner (1-30 microM). Mibefradil (30 microM) changed the pattern of pacemaker potentials from rapidly rising, high frequency events to slowly depolarizing, low-frequency events with considerable membrane noise (unitary potentials) between pacemaker potentials. Caffeine (3 mM) abolished pacemaker potentials in the presence of mibefradil. Pinacidil (10 microM), an ATP-sensitive K(+) channel opener, hyperpolarized ICC-MY and increased the amplitude and dV/dt(max) without affecting frequency. Pinacidil hyperpolarized smooth muscle cells and attenuated the amplitude and dV/dt(max) of slow waves without affecting frequency. The effects of pinacidil were blocked by glibenclamide (10 microM). These data suggest that slow waves are electrotonic potentials driven by pacemaker potentials. The upstroke component of pacemaker potentials is due to activation of dihydropyridine-resistant Ca(2+) channels, and this depolarization entrains pacemaker activity to create the plateau potential. The plateau potential may be due to summation of unitary potentials generated by individual or small groups of pacemaker units in ICC-MY. Entrainment of unitary potentials appears to depend on Ca(2+) entry during upstroke depolarization. PMID- 15537709 TI - Morphofunctional integration between skeletal myoblasts and adult cardiomyocytes in coculture is favored by direct cell-cell contacts and relaxin treatment. AB - The success of cellular cardiomyoplasty, a novel therapy for the repair of postischemic myocardium, depends on the anatomical integration of the engrafted cells with the resident cardiomyocytes. Our aim was to investigate the interaction between undifferentiated mouse skeletal myoblasts (C2C12 cells) and adult rat ventricular cardiomyocytes in an in vitro coculture model. Connexin43 (Cx43) expression, Lucifer yellow microinjection, Ca2+ transient propagation, and electrophysiological analysis demonstrated that myoblasts and cardiomyocytes were coupled by functional gap junctions. We also showed that cardiomyocytes upregulated gap junctional communication and expression of Cx43 in myoblasts. This effect required direct cell-to-cell contact between the two cell types and was potentiated by treatment with relaxin, a cardiotropic hormone with potential effects on cardiac development. Analysis of the gating properties of gap junctions by dual cell patch clamping showed that the copresence of cardiomyocytes in the cultures significantly increased the transjunctional current and conductance between myoblasts. Relaxin enhanced this effect in both the myoblast-myoblast and myoblast-cardiomyocyte cell pairs, likely acting not only on gap junction formation but also on the electrical properties of the preexisting channels. Our findings suggest that myoblasts and cardiomyocytes interact actively through gap junctions and that relaxin potentiates the intercellular coupling. A potential role for gap junctional communication in favoring the intercellular exchange of regulatory molecules, including Ca2+, in the modulation of myoblast differentiation is discussed. PMID- 15537710 TI - Enhanced response to caffeine and 4-chloro-m-cresol in malignant hyperthermia susceptible muscle is related in part to chronically elevated resting [Ca2+]i. AB - Malignant hyperthermia (MH) is a potentially fatal pharmacogenetic syndrome caused by exposure to halogenated volatile anesthetics and/or depolarizing muscle relaxants. We have measured intracellular Ca(2+) concentration ([Ca(2+)](i)) using double-barreled, Ca(2+)-selective microelectrodes in myoballs prepared from skeletal muscle of MH-susceptible (MHS) and MH-nonsusceptible (MHN) swine. Resting [Ca(2+)](i) was approximately twofold in MHS compared with MHN quiescent myoballs (232 +/- 35 vs. 112 +/- 11 nM). Treatment of myoballs with caffeine or 4 chloro-m-cresol (4-CmC) produced an elevation in [Ca(2+)](i) in both groups; however, the concentration required to cause a rise in [Ca(2+)](i) elevation was four times lower in MHS than in MHN skeletal muscle cells. Incubation of MHS cells with the fast-complexing Ca(2+) buffer BAPTA reduced [Ca(2+)](i), raised the concentration of caffeine and 4-CmC required to cause an elevation of [Ca(2+)](i), and reduced the amount of Ca(2+) release associated with exposure to any given concentration of caffeine or 4-CmC to MHN levels. These results suggest that the differences in the response of MHS skeletal myoballs to caffeine and 4 CmC may be mediated at least in part by the chronic high resting [Ca(2+)](i) levels in these cells. PMID- 15537711 TI - Mild sustained and intermittent hypoxia induce apoptosis in PC-12 cells via different mechanisms. AB - Episodic hypoxia, a characteristic feature of obstructive sleep apnea, induces cellular changes and apoptosis in brain regions associated with neurocognitive function. To investigate whether mild, intermittent hypoxia would induce more extensive neuronal damage than would a similar degree of sustained hypoxia, rat pheochromocytoma PC-12 neuronal cells were subjected to either sustained (5% O(2)) or intermittent (alternating 5% O(2) 35 min, 21% O(2) 25 min) hypoxia for 2 or 4 days. Quantitative assessment of apoptosis showed that while mild sustained hypoxia did not significantly increase cell apoptosis at 2 days (1.31 +/- 0.29 fold, n = 8; P = NS), a significant increase in apoptosis occurred after 4 days (2.25 +/- 0.4-fold, n = 8; P < 0.002), without increased caspase activation. Furthermore, caspase inhibition with the general caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK) did not modify sustained hypoxia-induced apoptosis. In contrast, mild, intermittent hypoxia induced significant increases in apoptosis at 2 days (3.72 +/- 1.43-fold, n = 8; P < 0.03) and at 4 days (4.57 +/- 0.82-fold, n = 8; P < 0.001) that was associated with enhanced caspase activity and attenuated by Z-VAD-FMK pretreatment. We conclude that intermittent hypoxia induces an earlier and more extensive apoptotic response than sustained hypoxia and that this response is at least partially dependent on caspase-mediated pathways. In contrast, caspases do not seem to play a role in sustained hypoxia-induced apoptosis. These findings suggest that different signaling pathways are involved in sustained and intermittent hypoxia-induced cell injury and may contribute to the understanding of differential brain susceptibility to sustained and intermittent hypoxia. PMID- 15537712 TI - Control of respiration and bioenergetics during muscle contraction. AB - (1)H-NMR experiments have determined intracellular O(2) consumption (Vo(2)) with oxymyoglobin (MbO(2)) desaturation kinetics in human calf muscle during plantar flexion exercise at 0.75, 0.92, and 1.17 Hz with a constant load. At the onset of muscle contraction, myoglobin (Mb) desaturates rapidly. The desaturation rate constant of approximately 30 s reflects the intracellular Vo(2). Although Mb desaturates quickly with a similar time constant at all workload levels, its final steady-state level differs. As work increases, the final steady-state cellular Po(2) decreases progressively. After Mb desaturation has reached a steady state, however, Vo(2) continues to rise. On the basis of current respiratory control models, the analysis in the present report reveals two distinct Vo(2) phases: an ADP-independent phase at the onset of contraction and an ADP-dependent phase after Mb has reached a steady state. In contrast to the accepted view, the initial intracellular Vo(2) shows that oxidative phosphorylation can support up to 36% of the energy cost, a significantly higher fraction than expected. Partitioning of the energy flux shows that a 31% nonoxidative component exists and responds to the dynamic energy utilization restoration cycle (which lasts for only milliseconds) as postulated in the glycogen shunt theory. The present study offers perspectives on the regulation of respiration, bioenergetics, and Mb function during muscle contraction. PMID- 15537713 TI - Histone deacetylase inhibition-mediated neuronal differentiation of multipotent adult neural progenitor cells. AB - It has become apparent that chromatin modification plays a critical role in the regulation of cell-type-specific gene expression. Here, we show that an inhibitor of histone deacetylase, valproic acid (VPA), induced neuronal differentiation of adult hippocampal neural progenitors. In addition, VPA inhibited astrocyte and oligodendrocyte differentiation, even in conditions that favored lineage-specific differentiation. Among the VPA-up-regulated, neuron-specific genes, a neurogenic basic helix-loop-helix transcription factor, NeuroD, was identified. Overexpression of NeuroD resulted in the induction and suppression of neuronal and glial differentiation, respectively. These results suggest that VPA promotes neuronal fate and inhibits glial fate simultaneously through the induction of neurogenic transcription factors including NeuroD. PMID- 15537714 TI - Eradication of American trypanosomiasis (Chagas' disease): an achievable goal? PMID- 15537715 TI - Bacterial isolates in the sick young infant in Ile-Ife, Nigeria. AB - This prospective study was carried out to determine the bacterial pathogens and their antibiotic sensitivity profile in the sick young infant. All consecutive young infants with features suggestive of infection seen over 5 months at a Comprehensive Health Centre in Ile-Ife, were screened for septicaemia and local bacterial infections. Of the 121 sick young infants screened for infection, 94 (77.7 per cent) had confirmed bacterial infection and 54 (57.4 per cent) of the 94 had confirmed septicaemia. Gram-positive organisms were the commonest bacterial isolates accounting for 204 (81.6 per cent) of the 250 isolates in this study. Staphylococcus aureus was the most frequent organism accounting for 61.2 per cent of all isolates. Gram-negative organisms accounted for 46 (18.4 per cent) of all isolates with Salmonella spp. and Proteus vulgaris predominating. All the bacterial isolates in this study were sensitive to ofloxacin and most were sensitive to the antibiotics commonly employed in the treatment of infections caused by these organisms. However, many of both Grampositive and Gram negative isolates in this study were resistant to cotrimoxazole. The study highlights the high prevalence of bacterial infections (localized or systemic) among young infants. It also shows that Gram-positive organisms, the principal aetiologic agents, were sensitive to commonly used antibiotics. It is recommended that genticin and cloxacillin or erythromycin should be used as the first-line antibiotics in the treatment of young infants with bacterial infections in Ile Ife, Nigeria. PMID- 15537716 TI - Peptic ulcer disease in children and adolescents. AB - Peptic ulcer disease (PUD) has been reported to occur in children worldwide, but no information is available for our community. The aim of the study was to report our experience on the pattern of this condition in Saudi Arabian children. The records of all children below 18 years of age who were diagnosed by endoscopy to have PUD over a period of 10 years were analysed. From 1993 to 2002, 24 children out of 521(5 per cent) who presented with upper gastrointestinal tract (GIT) symptoms were diagnosed by endoscopy to have PUD. All but one (96 per cent) were Saudi nationals, the average age was 15 years (range 5-18 years), and the male to female ratio was 7:1. The commonest presentation was chronic abdominal pain in 15/24 (63 per cent) of the children, followed by vomiting associated with abdominal pain in four (17 per cent). Hematemesis and melena occurred in three (13 per cent), and two children (8 per cent), respectively. There were 20 duodenal (92 per cent) and four gastric ulcers. The primary type was the most common, occurring in 19 (79 per cent) of the children. Histopathology results of antral biopsies were available for 15 children; all of them had antral gastritis. Helicobacter pylori organisms were present in 13/15 (87 per cent) of the antral biopsy specimens. In Saudi children, peptic ulcer disease occurs more commonly in boys. It is a rare cause of upper GIT symptoms, but highly associated with H. pylori antral gastritis. This study documents a pattern similar to descriptions from other countries. PMID- 15537717 TI - Level of awareness of the baby friendly initiative among home economics teachers in Nigeria. AB - A cross-sectional questionnaire was used to assess the level of awareness of the Baby Friendly Initiative among Home Economics teachers in Nigeria. Only 34.5 per cent of the teachers were able to define BFI as the Baby Friendly Initiative while 60 per cent defined exclusive breastfeeding correctly. Although 70.2 per cent acknowledged that breastfeeding prevents malnutrition in babies, only 12.5 per cent knew about the protective effects of colostrum. Of the study population 29.8 per cent knew that breastfeeding should be on demand and not regulated. Over 80 per cent of the teachers advocated that the BFI should be taught at all levels of education. We conclude that their knowledge of the BFI is poor and that they will require further education before they can be used effectively as resource personnel in training girls (potential mothers) in BFI. PMID- 15537718 TI - The Comparison of PRISM and PIM scoring systems for mortality risk in infantile intensive care. AB - Scoring systems that predict the risk of mortality for children in an intensive care unit (ICU) are needed for the evaluation of the effectiveness of pediatric intensive care. The Pediatric Risk of Mortality (PRISM) and the Pediatric Index of Mortality (PIM) scores have been developed to predict mortality among children in the ICU. The purpose of this study was to evaluate whether these systems are effective and population-independent. PRISM and PIM scores were calculated prospectively during a 1-year period solely on 105 non-surgical infants admitted to the ICU. Statistical analysis was performed to assess the performance of the scoring systems. There were 29 (27.6 per cent) deaths and 76 (72.4 per cent) survivors. SMR and Z scores for PIM and PRISM signified higher mortality and poor performance. Prediction of mortality by the scoring systems appeared to be underestimated in almost all risk groups. The Hosmer and Lemeshow test showed a satisfactory overall calibration of both scoring systems. Although ROC analysis showed a poor discriminatory function of both scores, a marginally acceptable performance for PIM was observed. The ROC curve also showed an acceptable performance for PIM, for patients with pre-existent chronic disorder. Although care must be taken not to overstate the importance of our results, we believe that when revised according to the characteristics of the population, PIM may perform well in predicting the mortality risk for infants in the ICU, especially in countries where the mortality rate is relatively high and pre-existent chronic disorders are more common. PMID- 15537719 TI - Evaluation of early detection and management of disseminated intravascular coagulation among Alexandria University pediatric intensive care patients. AB - This prospective study over 24 months aimed to evaluate the outcome of early management of disseminated intravascular coagulation (DIC) among high-risk patients (n = 50) admitted to a pediatric intensive care unit (PICU). It also included all cases presenting with overt DIC (OD) concomitantly (n = 30). The high-risk group (pre-DIC) was subdivided, according to their D-dimer assay, into negative (n = 14) and positive (n = 36) D-dimer groups. All three groups were evaluated, on admission, for their prothrombin time (PT), activated partial thromboplastin time (APTT), plasma fibrinogen level (Fi), fibrinogen degradation products (FDP), platelet count, and presence/absence of schistocytes in peripheral blood. The combination of D-dimer and FDP assay showed the best correlation for early pre-DIC diagnosis (r = 0.9048). FDP assay was the best parameter for followup of progress of DIC condition in the PICU. The lowest mortality was among negative D-dimer, followed by positive D-dimer and OD groups (28.6 per cent, 77.8 per cent, and 93.3 per cent, respectively). Among the positive D-dimer group the lowest mortality was encountered in the subgroup treated with plasma, heparin and tranexamic acid (33 per cent) while those treated with non-specific therapy, plasma only, or plasma and heparin showed higher mortality (100 per cent, 80 per cent, and 100 per cent, respectively). The deceased subgroup, among positive D-dimer cases showed a significantly higher number of patients presenting with multiple organ failure on admission compared with the discharged group. In summary, early diagnosis and proper management of pre-DIC, before overt bleeding, in high-risk patients admitted to a PICU using combined D-dimer and FDP assays had a positive impact on their prognosis. PMID- 15537720 TI - Towards a better understanding of exclusive breastfeeding in the era of HIV/AIDS: a study of prevalence and factors associated with exclusive breastfeeding from birth, in Rakai,Uganda. AB - Exclusive breastfeeding (EBF) is being promoted by WHO/UNICEF as the recommended mode of feeding for all infants from birth to around 6 months of age. It is also recommended for the initial few months, for infants born to HIV-positive mothers in the developing world where conditions may not be ideal for replacement feeding. A cross-sectional study was done to find the prevalence and factors associated with EBF from delivery, amongst mothers of infants 0-6 months of age in a rural Ugandan community with a high HIV/AIDS burden. A prevalence of EBF of 35.1 per cent was found. There was a very high use of prelacteal feeds. Factors positively associated with EBF from birth, after logistic regression, were: delivery from a health unit (OR 2.07; 95 per cent confidence interval, 1.2-3.6); and mother having a normal vaginal delivery (OR 10.54; 95 per cent CI, 3.0-36.6). Factors showing negative association were: male gender of child (OR 0.44; 95 per cent CI, 0.3-0.7); and age of child over 3 months (OR 0.48; 95 per cent CI, 0.3 0.7). Mothers knowledge of her HIV serostatus was not associated with EBF (p = 0.78). Breastfeeding is almost universal in this area but it is rarely exclusive from birth. PMID- 15537721 TI - Plesiomonas shigelloides-associated diarrhoea in Bangladeshi children: a hospital based surveillance study. AB - Little is known about clinical and epidemiological features of Plesiomonas shigelloides-associated diarrhoea in children. We reviewed hospital-based surveillance records of 38 children with diarrhoea having P. shigelloides as the only pathogen isolated from their faecal specimen. Of those 38 children, 29 (76 per cent) were below 2 years of age and 28 (74 per cent) were male. Thirty-two (84 per cent) children presented with watery diarrhoea and six (16 per cent) had dysenteric stools. Vomiting was a feature in 27 (71 per cent) children and clinically significant dehydration was observed in nine (23 per cent) children. Fever was present in three (8 per cent) children and five (13 per cent) had diarrhoea 14 days. Thirty-three (87 per cent) children were successfully treated with ORS alone and only five (13 per cent) required intravenous fluid in addition to ORS. Plesiomonas shigelloides was isolated throughout the year. The findings may be of public health importance for creating awareness among physicians about the clinical profile and management strategy of P. shigelloides-associated diarrhoea in children. PMID- 15537722 TI - Antioxidant micronutrient profile (vitamin E, C, A, copper, zinc, iron) of colostrum: association with maternal characteristics. AB - The study was conducted to investigate the micronutrient profile of human colostrum, and to assess the association of maternal characteristics to the micronutrients. Colostral concentrations of antioxidant vitamins E, C, and A were 21.34 +/- 8.47, 148.92 +/- 43.64, 0.79 +/- 0.42 micromol/l, respectively. The antioxidant minerals copper, zinc, and iron contents were 19.17 +/- 11.73, 63.69 +/- 12.82, 11.44 +/- 1.46 micromol/l, respectively. Maternal characteristics did not have any influence on the colostral micronutrients. PMID- 15537723 TI - First study of CF mutations in the CFTR gene of Iranian patients: detection of DeltaF508, G542X, W1282X, A120T, R117H, and R347H mutations. AB - Thirty-seven unrelated Iranian CF families were screened for the presence of seven common mutations (DeltaF508, G542X, W1282X, G551D, N1303K, 1717-1G-->A, and 621-1G-->T) using ARMS PCR and exons 4 and 7 of the CFTR gene by SSCP method. This study resulted in the identification of 26.8 per cent of all CF alleles: DeltaF508 (16.2 per cent), W1282X (4 per cent), G542X (2.7 per cent), R117H (1.3 per cent), R347H (1.3 per cent), and A120T (1.3 per cent) mutations were detected. To the best of our knowledge, it is the first report of an Asian subject carrying the A120T mutation. Our findings suggest heterogeneity in the Iranian population, stressing the need to draw attention to sequence analysis in order to find population-specific mutations. PMID- 15537724 TI - Calvarial tuberculosis. AB - Tuberculosis of the skull is very rare. We report on two children: one presented with orbital and scalp swellings and was found to have lytic lesions on skull X ray, while the other presented with chronic discharging scalp ulcers typical of tuberculosis of the scalp and also showed lytic skull lesions. Both patients had evidence of vertebral and lung tuberculosis. PMID- 15537725 TI - The frequency of nutritional rickets among hospitalized infants and its relation to respiratory diseases. AB - This case control study was conducted to determine the frequency of nutritional rickets among hospitalized infants and to assess their relation to respiratory diseases. All infants between the age of 3 months and 2 years admitted to the pediatric ward of Queen Alia Military Hospital during the period February-October 2001 were examined and investigated to rule out nutritional rickets. Children admitted for the first time to hospital for acute illnesses were only included in the study. A special data collection sheet was designed for this study which includes information on the age, sex, causes of admission, family size, the rank of the child in the family, family monthly income, outdoor clothing habit of the mother, and the mode of feeding. Data were collected from the infant charts and/or by interviewing the child's mother or guardian. Clinical signs of rickets were also recorded, including rosary beads, craniotabes, wide anterior fontanel, delayed dentition, widening of epiphysis, bowing of the legs, and double malulous. Blood sample was collected for calcium, phosphorus, alkaline phosphatase, and hemoglobin level. Those infants with any clinical sign of rickets and/or abnormal chemical results had a wrist X-ray to confirm the diagnosis of rickets. The rachitic group (cases) was compared for statistical significance with the remaining non-rachitic infants (controls) for the data collected. Rachitic infants received intramuscular 600,000 IU of vitamin D; a follow-up wrist X-ray and blood sample for calcium, phosphorus and alkaline phosphatase was arranged 3 weeks later. Forty-seven infants (10.6 per cent) out of the 443 included in the study were found to have nutritional rickets. Forty (85.1 per cent) of the rachitic infants were admitted due to lower respiratory tract diseases compared with 30 per cent of the control group and the difference was statistically significant (p < 0.01). Duration of hospital stay in the rachitic infants was also significantly more prolonged than the non-rachitic control group (9.5 days vs. 7.4 days, p = 0.002). Rachitic infants were breastfed in 82.9 per cent, ranked second or more in the family in 87.2 per cent, and had mothers who wore head cover outdoors in 80.8 per cent compared with 60.8, 40.1, and 60.3 per cent, respectively, in the non-rachitic group (p < 0.01). High alkaline phosphatase, hypocalcemia, hypophosphatemia, and anemia was found in 100, 19, 50, and 78.7 per cent, respectively, in the rachitic group compared with 9.8, 2, 1.2, and 43.7 per cent, respectively, in the control group (p < 0.001). Nutritional rickets seems to be a common problem among infants in Jordan. Further studies at national level are needed to determine the prevalence of rickets in Jordan. Rachitic infants are commonly hospitalized due to lower respiratory tract infections, thus there is a high index of suspicion for rickets among hospitalized infants with lower respiratory tract diseases. PMID- 15537726 TI - Serum transferrin receptor levels in beta-thalassemia trait. AB - Increased levels of transferrin receptors (TfR) and soluble TfR (sTfR) are reliable indicators of iron deficiency anemia and conditions with enhanced erythropoietic activity. In this study we measured sTfR levels in the cases of beta-thalassemia trait (betaTT) and IDA to see whether it would be a diagnostic tool for the differentiation of betaTT and IDA. On the other hand STfR levels could give an idea about erythropoietic activity in betaTT. Serum transferrin receptor (sTfR) levels were measured with enzyme immune assay in 37 cases with betaTT, in 26 patients with iron deficiency anemia (IDA), in 17 patients with the coexistence of betaTT with IDA, and in 21 healthy controls. Mean sTfR in betaTT (6.54 +/- 1.88) were statistically higher than in controls (5.09 +/- 0.76) and lower than in IDA (11.46 +/- 5.03) and in IDA coexistence with betaTT (14.44 +/- 5.3, p < 0.05 in each comparison). However, in 17 cases with betaTT sTfR levels showed an overlap between the patients with IDA, and in 15 cases with betaTT it also showed an overlap with controls. The finding of higher sTfR levels in betaTT compared with controls suggested that there was a degree of ineffective hematopoiesis in betaTT. Although sTfR levels in IDA were higher than in betaTT, because of largely overlapping values in both groups, it was not satisfactory to distinguish betaTT from IDA. PMID- 15537727 TI - An uncommon presentation: chronic meningococcaemia associated with cholestatic hepatitis in a Turkish child. AB - Chronic meningococcaemia is a very rare clinical manifestation of invasive infection by Neisseria meningitidis. A 9-year-old girl was admitted to our clinic with complaints of fever, headache, arthralgia, and maculopapular rash. The diagnosis was made by the growth of Neisseria meningitidis in the blood cultures. Four days after admission, liver function tests were increased and were compatible with cholestatic hepatitis. Thereafter, the patient was successfully treated and symptoms were completely resolved. To our knowledge, there have been no previous reports of Neisseria meningitidis causing cholestatic hepatitis. Herein, we present an unusual child patient with chronic meningococcaemia associated with cholestatic hepatitis. PMID- 15537728 TI - Multiple organ involvement hydatid cysts in a 2-year-old boy. AB - The cerebral hydatid cysts caused by Echinococcus granulosus are rare and occur mostly during childhood in endemic areas. A 2-year-old boy was admitted with focal neurological signs in the left extremities. Magnetic resonance imaging of the brain showed a cyst lying from right parietooccipital region to the lateral ventricle. There were also multiple cysts in his lung and liver. The cerebral hydatid cyst was surgically extracted without complications. We suggest that a differential diagnosis of hydatid cyst should be considered when a brain mass is found in a patient, even 2 years old, from an endemic area of echinococcosis. PMID- 15537729 TI - Congenital trypanosomiasis. AB - The last successfully treated case of congenital trypanosomiasis in Zambia was in October 1978, with detailed analysis of immunoglobulins, illustrating the waning of blood and serum levels of IgA, IgG, and IgM during treatment, up to 99 days after treatment. Twenty-five years later, we report on a case of congenital trypanosomiasis. The disease is now rare and can be missed or dismissed as retroviral disease, particularly in adults. The main unusual symptoms were the prolonged intermittent convulsions in an otherwise well infant. Management of the disease is now more interdisciplinary, resources for laboratory support are fewer, lumbar puncture is more relevant, and antitrypanosomal drugs are more difficult to obtain. The mother died within one week of hospitalization and the infant initially responded to three doses of suramin and 3 weeks of melsopropol. Convulsions ceased during the second round of melsopropol. Unfortunately, the infant died of nosocomial infection. PMID- 15537730 TI - MBL deficiency does not associate with recurrent pneumococcal infections in a group of Colombian children. PMID- 15537731 TI - Retreatment of long-lasting insecticide-treated mosquito nets under field conditions in rural Burkina Faso. PMID- 15537732 TI - A semi-persistent adult ocular dominance plasticity in visual cortex is stabilized by activated CREB. AB - The adult cerebral cortex can adapt to environmental change. Using monocular deprivation as a paradigm, we find that rapid experience-dependent plasticity exists even in the mature primary visual cortex. However, adult cortical plasticity differs from developmental plasticity in two important ways. First, the effect of adult, but not juvenile monocular deprivation is strongly suppressed by administration of barbiturate just prior to recording visual evoked potentials, suggesting that the effect of adult experience can be inactivated acutely. Second, the effect of deprivation is less persistent over time in adults than in juveniles. This correlates with the known decline in CREB function during maturation of the visual cortex. To compensate for this decline in CREB function, we expressed persistently active VP16-CREB and find that it causes adult plasticity to become persistent. These results suggest that in development and adulthood, the regulation of a trans-synaptic signaling pathway controls the adaptive potential of cortical circuits. PMID- 15537733 TI - Central localization of plasticity involved in appetitive conditioning in Lymnaea. AB - Learning to associate a conditioned (CS) and unconditioned stimulus (US) results in changes in the processing of CS information. Here, we address directly the question whether chemical appetitive conditioning of Lymnaea feeding behavior involves changes in the peripheral and/or central processing of the CS by using extracellular recording techniques to monitor neuronal activity at two stages of the sensory processing pathway. Our data show that appetitive conditioning does not affect significantly the overall CS response of afferent nerves connecting chemosensory structures in the lips and tentacles to the central nervous system (CNS). In contrast, neuronal output from the cerebral ganglia, which represent the first central processing stage for chemosensory information, is enhanced significantly in response to the CS after appetitive conditioning. This demonstrates that chemical appetitive conditioning in Lymnaea affects the central, but not the peripheral processing of chemosensory information. It also identifies the cerebral ganglia of Lymnaea as an important site for neuronal plasticity and forms the basis for detailed cellular studies of neuronal plasticity. PMID- 15537734 TI - Classical conditioning of eyelid and mystacial vibrissae responses in conscious mice. AB - The murine vibrissae sensorimotor system has been scrutinized as a target of motor learning through trace classical conditioning. Conditioned eyelid responses were acquired by using weak electrical whisker-pad stimulation as conditioned stimulus (CS) and strong electrical periorbital stimulation as unconditioned stimulus (US). In addition, conditioned vibrissal protraction was obtained pairing either weak electrical whisker-pad stimulation or a tone as CS, with a strong electric shock delivered in the whisker-pad as US. This finding suggests that evolutionary pressure has selected a sensorimotor system capable of constructing conditioned responses on the basis of temporal relationships of stimuli, independently of any putative functional purpose. PMID- 15537735 TI - Coordinate high-frequency pattern of stimulation and calcium levels control the induction of LTP in striatal cholinergic interneurons. AB - Current evidence appoints a central role to cholinergic interneurons in modulating striatal function. Recently, a long-term potentiation (LTP) of synaptic transmission has been reported to occur in these neurons. The relationship between the pattern of cortico/thalamostriatal fibers stimulation, the consequent changes in the intracellular calcium concentration ([Ca2+]i), and the induction of synaptic plasticity was investigated in striatal cholinergic interneurons from a rat corticostriatal slice preparation by means of combined electrophysiological intracellular recordings and microfluorometric techniques. Different protocols of stimulation were considered, varying both the frequency and the duration of the train of stimuli. High-frequency stimulation (HFS) (three trains at 100 Hz for 3 sec, 20-sec interval) induced a rise in [Ca2+]i, exceeding by fivefold the resting level, and caused a LTP of synaptic transmission. Tetanic stimulation delivered at lower frequencies (5-30 Hz) failed to induce long-term changes of synaptic efficacy. The observed elevation in [Ca2+]i during HFS was primarily mediated by L-type high-voltage activated (HVA)-Ca2+ channels, as it was fully prevented by nifedipine. Conversely, blockade of NMDA and AMPA glutamate receptor did not affect either LTP or the magnitude of the [Ca2+]i rise. Interestingly, the pharmacological analysis of the post-tetanic depolarizing postsynaptic potential (DPSP) revealed that LTP was attributable, to a large extent, to the potentiation of the GABA(A)-mediated component. In conclusion, the expression of LTP in striatal cholinergic interneurons is a selective response to a precise stimulation pattern of induction requiring a critical rise in [Ca2+]i. PMID- 15537736 TI - Impaired odor recognition memory in patients with hippocampal lesions. AB - In humans, impaired recognition memory following lesions thought to be limited to the hippocampal region has been demonstrated for a wide variety of tasks. However, the importance of the human hippocampus for olfactory recognition memory has scarcely been explored. We evaluated the ability of memory-impaired patients with damage thought to be limited to the hippocampal region to recognize a list of odors. The patients were significantly impaired after a retention delay of 1 h. Olfactory sensitivity was intact. This finding is in agreement with earlier reports that rats with hippocampal lesions exhibited memory impairment on an odor delayed nonmatching to sample task (after 30 min and 1 h) and that patients with damage thought to be limited to the hippocampal region were impaired on an odor span memory task. Olfactory recognition memory, similar to recognition memory in other sensory modalities, depends on the integrity of the hippocampal region. PMID- 15537737 TI - Cerebellar norepinephrine modulates learning of delay classical eyeblink conditioning: evidence for post-synaptic signaling via PKA. AB - The neurotransmitter norepinephrine (NE) has been shown to modulate cerebellar dependent learning and memory. Lesions of the nucleus locus coeruleus or systemic blockade of noradrenergic receptors has been shown to delay the acquisition of several cerebellar-dependent learning tasks. To date, no studies have shown a direct involvement of cerebellar noradrenergic activity nor localized the post synaptic response to cerebellar beta-noradrenergic receptor signaling. Using ipsilateral, localized infusions into cerebellar lobule HVI and interpositus (IP), we have established that blocking beta-noradrenergic receptors with propranolol significantly impairs acquisition of conditioned responses. Furthermore, interrupting activation of cAMP-dependent PKA in the cerebellum using Rp-cAMPS completely prevents acquisition. However, neither blocking beta adrenergic receptors nor blocking PKA activation significantly interferes with performance of established conditioned responses when administered after the learned response is formed. PMID- 15537738 TI - Differential regulation of brain-derived neurotrophic factor transcripts during the consolidation of fear learning. AB - Brain-derived neurotrophic factor (BDNF) has been implicated as a molecular mediator of learning and memory. The BDNF gene contains four differentially regulated promoters that generate four distinct mRNA transcripts, each containing a unique noncoding 5'-exon and a common 3'-coding exon. This study describes novel evidence for the differential usage of alternative BDNF promoters and 5' exons during the consolidation of learning. We found a selective increase in BDNF transcripts containing exons I and III in the amygdala 2 h following fear conditioning, while mRNA levels of BDNF exons II and IV remained unchanged. These results provide the first evidence of differential splicing and/or differential BDNF promoter usage in response to a behaviorally relevant learning paradigm. PMID- 15537739 TI - Olfactory fear conditioning induces field potential potentiation in rat olfactory cortex and amygdala. AB - The widely used Pavlovian fear-conditioning paradigms used for studying the neurobiology of learning and memory have mainly used auditory cues as conditioned stimuli (CS). The present work assessed the neural network involved in olfactory fear conditioning, using olfactory bulb stimulation-induced field potential signal (EFP) as a marker of plasticity in the olfactory pathway. Training consisted of a single training session including six pairings of an odor CS with a mild foot-shock unconditioned stimulus (US). Twenty-four hours later, the animals were tested for retention of the CS as assessed by the amount of freezing exhibited in the presence of the learned odor. Behavioral data showed that trained animals exhibited a significantly higher level of freezing in response to the CS than control animals. In the same animals, EFPs were recorded in parallel in the anterior piriform cortex (aPC), posterior piriform cortex (pPC), cortical nucleus of the amygdala (CoA), and basolateral nucleus of the amygdala (BLA) following electrical stimulation of the olfactory bulb. Specifically, EFPs recorded before (baseline) and after (during the retention test) training revealed that trained animals exhibited a lasting increase (present before and during presentation of the CS) in EFP amplitude in CoA, which is the first amygdaloid target of olfactory information. In addition, a transient increase was observed in pPC and BLA during presentation of the CS. These data indicate that the olfactory and auditory fear-conditioning neural networks have both similarities and differences, and suggest that the fear-related behaviors in each sensory system may have at least some distinct characteristics. PMID- 15537740 TI - Long-term memory for instrumental responses does not undergo protein synthesis dependent reconsolidation upon retrieval. AB - Recent evidence indicates that certain forms of memory, upon recall, may return to a labile state requiring the synthesis of new proteins in order to preserve or reconsolidate the original memory trace. While the initial consolidation of "instrumental memories" has been shown to require de novo protein synthesis in the nucleus accumbens, it is not known whether memories of this type undergo protein synthesis-dependent reconsolidation. Here we show that low doses of the protein synthesis inhibitor anisomycin (ANI; 5 or 20 mg/kg) administered systemically in rats immediately after recall of a lever-pressing task potently impaired performance on the following daily test sessions. We determined that the nature of this impairment was attributable to conditioned taste aversion (CTA) to the sugar reinforcer used in the task rather than to mnemonic or motoric impairments. However, by substituting a novel flavored reinforcer (chocolate pellets) prior to the administration of doses of ANI (150 or 210 mg/kg) previously shown to cause amnesia, a strong CTA to chocolate was induced sparing any aversion to sugar. Importantly, when sugar was reintroduced on the following session, we found that memory for the task was not significantly affected by ANI. Thus, these data suggest that memory for a well-learned instrumental response does not require protein synthesis-dependent reconsolidation as a means of long term maintenance. PMID- 15537741 TI - Demonstration of nondeclarative sequence learning in mice: development of an animal analog of the human serial reaction time task. AB - In this paper, we demonstrate nondeclarative sequence learning in mice using an animal analog of the human serial reaction time task (SRT) that uses a within group comparison of behavior in response to a repeating sequence versus a random sequence. Ten female B6CBA mice performed eleven 96-trial sessions containing 24 repetitions of a 4-trial sequence. During the 12th session, the repeating sequence was replaced with the random sequence halfway through the session. Reaction time (RT) to respond to an illuminated nose-poke was recorded, and performance was compared at the halfway point in each session to test for any change in behavior. For learning effect, RTs decreased over the no-switch repeating-sequence sessions. For interference effect, behavior did not change appreciably at the halfway point during the last repeating-sequence session. However, RTs deteriorated significantly after the switch from repeating to random sequences halfway through session 12. The mice demonstrated a robust interference effect when switched from repeating to random sequences. This pattern of behavior in humans performing the SRT is interpreted as evidence of nondeclarative sequence learning. The similarity between the human and mouse SRTs will enable more direct comparisons of mouse-human nondeclarative memory behavior and will provide a useful behavioral end-point in mouse-models of basal ganglia dysfunction. PMID- 15537742 TI - Nonassociative learning processes determine expression and extinction of conditioned fear in mice. AB - Freezing to a tone following auditory fear conditioning is commonly considered as a measure of the strength of the tone-shock association. The decrease in freezing on repeated nonreinforced tone presentation following conditioning, in turn, is attributed to the formation of an inhibitory association between tone and shock that leads to a suppression of the expression of fear. This study challenges these concepts for auditory fear conditioning in mice. We show that acquisition of conditioned fear by a few tone-shock pairings is accompanied by a nonassociative sensitization process. As a consequence, the freezing response of conditioned mice seems to be determined by both associative and nonassociative memory components. Our data suggest that the intensity of freezing as a function of footshock intensity is primarily determined by the nonassociative component, whereas the associative component is more or less categorical. We next demonstrate that the decrease in freezing on repeated nonreinforced tone presentation following conditioning shows fundamental properties of habituation. Thus, it might be regarded as a habituation-like process, which abolishes the influence of sensitization on the freezing response to the tone without affecting the expression of the associative memory component. Taken together, this study merges the dual-process theory of habituation with the concept of classical fear conditioning and demonstrates that sensitization and habituation as two nonassociative learning processes may critically determine the expression of conditioned fear in mice. PMID- 15537743 TI - Interaction of polycyclic musks and UV filters with the estrogen receptor (ER), androgen receptor (AR), and progesterone receptor (PR) in reporter gene bioassays. AB - Two important ingredients of personal care products, namely polycyclic musk fragrances and UV filters, can be found in the environment and in humans. In previous studies, several compounds of both classes have been tested for their interaction with the estrogen receptor. Two polycyclic musk fragrances, namely AHTN and HHCB, turned out to be anti-estrogenic both in vitro and in vivo in a transgenic zebrafish assay. Several UV filters have been shown to exert estrogenic effects in vitro and in some in vivo studies. Here, we assessed the interaction of five polycyclic musk compounds and seven UV filters with the estrogen receptor (ER), androgen receptor (AR), and progesterone (PR) receptor, using sensitive and specific reporter gene cell lines. Four polycyclic musks (AHTN, HHCB, AETT, and AHMI) were found to be antagonists toward the ERbeta, AR and PR. The UV filters that showed estrogenic effects (benzophenone-3, Bp-3; 3 benzylidene camphor, 3-BC; homosalate, HMS; and 4-methylbenzylidene camphor, 4 MBC) were found to be antagonists toward the AR and PR. The ERalpha agonistic UV filter octyl-dimethyl-p-aminobenzoic acid (OD-PABA) did not show activity toward the AR and PR. Octyl methoxy cinnamate (OMC) showed weak ERalpha agonism, but potent PR antagonism. Butyl methoxydibenzoylmethane (B-MDM) only showed weak ERalpha agonism and weak AR antagonism. Most effects were observed at relatively high concentrations (above 1 muM); however, the anti-progestagenic effects of the polycyclic musks AHMI and AHTN were detected at concentrations as low as 0.01 muM. The activity of anti-progestagenic xenobiotics at low concentrations indicates the need to undertake more research to find out about the potential endocrine disrupting effects of these compounds in vivo. PMID- 15537744 TI - Acute hemorrhagic myocardial necrosis and sudden death of rats exposed to a combination of ephedrine and caffeine. AB - Because of possible side effects of herbal medicines containing ephedrine and guarana-derived caffeine, including increased risk of stroke, myocardial infarction, and sudden death, the Food and Drug Administration recently banned the sale of ephedra-containing products, specifically over-the-counter dietary supplements. We report cardiac in 7- and 14-week-old male F344 rats exposed by gavage to ephedrine(25 mg/kg) and caffeine (30 mg/kg) administered in combination for one or two days. The ephedrine-caffeine dosage was approximately 12- and 1.4 fold, respectively, above average human exposure, based on a mg/m2 body surface area comparison. Several (5/7) of the exposed 14-week-old rats died or were sacrificed in extremis 4-5 h after the first dosing. In these hearts, changes were observed chiefly in the interventricular septum but also left and right ventricular walls. Massive interstitial hemorrhage, with degeneration of myofibers, occurred at the subendocardial myocardium of the left ventricle and interventricular septum. Immunostaining for cleaved caspase-3 and hyperphosphorylated H2A.X, a histone variant that becomes hyperphosphorylated during apoptosis, indicated multifocal generalized positive staining of degenerating myofibers and fragmenting nuclei, respectively. The Barbeito-Lopez trichrome stain revealed generalized patchy yellow myofibers consistent with degeneration and/or coagulative necrosis. In ephedrine-caffeine-treated animals terminated after the second dosing, foci of myocardial degeneration and necrosis were already infiltrated by mixed inflammatory cells. The myocardial necrosis may occur secondarily to intense diffuse vasoconstriction of the coronary arterial system with decreased myocardial perfusion. Our work shows the direct relationship between combined ephedrine and caffeine exposure and cardiac pathology. PMID- 15537745 TI - Effects of co-administration of antioxidants and arsenicals on the rat urinary bladder epithelium. AB - Oxidative stress has been increasingly recognized as a possible mechanism in the toxicity and carcinogenicity of various chemicals, including arsenic. Therefore, treatment with antioxidants may afford a protective effect against arsenic induced cytotoxicity and carcinogenesis. Dimethylarsinic acid (DMAV) has been shown to be a bladder carcinogen in rats when administered at high doses (100 ppm) in the diet or in the drinking water. The main purpose of the present study was to evaluate the effects of co-administration of antioxidants with arsenicals on the rat urinary bladder epithelium in vitro and in vivo. In a previous experiment, treatment with 1000 ppm melatonin for two weeks did not inhibit cell proliferation induced in the rat urothelium by 100 ppm DMAV. In the current study, we examined the effects of five antioxidants that act via different mechanisms, on the in vitro cytotoxicity of various arsenicals, for the purpose of determining which antioxidants might have protective effects against arsenic induced cytotoxicity. The antioxidants that inhibited cytotoxicity in vitro were then studied also in vivo. Melatonin showed slight inhibition of the cytotoxicity of arsenite, but had no effect on the other arsenicals. N-acetylcysteine (NAC) inhibited the cytotoxicity of monomethylarsonous acid (MMAIII), DMAV, dimethylarsinous acid (DMAIII), and trimethylarsine oxide (TMAO). Vitamin C inhibited cytotoxicity induced by arsenate, arsenite, MMAIII) and DMAIII. Tiron and Trolox had no effect on the cytotoxicity of any arsenical. The in vitro inhibitory effects of NAC and vitamin C on DMAV and on DMAIII, suggested that these antioxidants might afford preventive effects on DMAV-induced bladder cytotoxicity and carcinogenesis in rats. To test this hypothesis, a 10-week rat bioassay was conducted. Melatonin was also included to clarify the results of the previous two-week experiment. The sodium salt of vitamin C (Na-Asc), but not melatonin or NAC, inhibited the proliferative effects of DMAV on the bladder epithelium in rats. These results suggest that oxidative stress is at least in part involved in DMAV-induced rat bladder toxicity and proliferation, and therefore, vitamin C may afford inhibitory effects in DMAV-induced bladder carcinogenesis in rats. Microarray analysis of DMAV-responsive genes revealed that DMAV did not have a consistent modifying effect on gene expression in the rat bladder epithelium, suggesting that proteins and/or lipids may be the targets of damage by DMAV-induced oxidative stress. PMID- 15537746 TI - Sodium arsenite exposure alters cell migration, focal adhesion localization and decreases tyrosine phosphorylation of focal adhesion kinase in H9C2 myoblasts. AB - Exposure to the environmental toxicant arsenic is reported to produce a variety of effects including disruption of signal transduction pathways, cell proliferation, and apoptosis. This suggests that arsenite may not have specific targets but rather extremely broad effects. The present study was designed to test the hypothesis that arsenite alters signaling involved in focal adhesion structure and function in cultured myoblasts. H9C2 cells were exposed to 1, 2.5, 5, or 10 microM sodium arsenite for 48 h. MTT metabolism and staining by neutral red, trypan blue, and propidium iodide showed that sodium arsenite treatments of 5 microM or less were not overtly cytotoxic. At these doses, sodium arsenite did not affect the amount of polymerized actin in cells, rate of protein synthesis, or amounts of vinculin, talin, paxillin, and focal adhesion kinase (FAK) in cells. However, sodium arsenite-treated cells contained fewer focal adhesions with an altered distribution pattern. Sodium arsenite exposure caused a dose dependent reduction in cell migration and cell attachment rates. The average area of substrate covered by a cell was also reduced, although the average volume of cells was not significantly affected. Sodium arsenite exposure resulted in reduced tyrosine phosphorylation of FAK, its substrate paxillin and the FAK auto- phosphorylation site, Tyr397. Our results indicate that sodium arsenite can alter focal adhesion structure and function, thus affecting cell attachment and migration and possibly other aspects of focal adhesion function such as integrin signaling. These diverse consequences may be mediated by a relatively specific inhibition of FAK tyrosine phosphorylation, modifying scaffolding proteins. PMID- 15537747 TI - Aryl hydrocarbon receptor expression and activity in cerebellar granule neuroblasts: implications for development and dioxin neurotoxicity. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent teratogen that produces neurobehavioral abnormalities associated with both cognitive and locomotor systems, yet the precise regional and cellular targets of developmental neurotoxicity remain largely unknown. Most, if not all, TCDD-induced pathology is mediated via binding to the aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor that belongs to the basic helix-loop-helix/Per-Arnt-Sim (bHLH/PAS) superfamily. Upon ligand binding, AhR translocates to the nucleus, dimerizes with the AhR nuclear translocator protein (Arnt), and regulates transcription by interaction with dioxin-response elements (DREs) in target genes, most notably specific cytochrome P450 (CYP) family members. To assess whether developing cerebellar granule neuroblasts are potential direct targets for TCDD toxicity, AhR expression and transcriptional activity were examined. AhR and Arnt proteins were present in mouse cerebellum from birth throughout postnatal development. AhR protein levels peaked between postnatal day (PND) 3 10, a critical period for granule neuroblast growth and maturation. Transcriptionally active AhR was detected in immature cerebellar granule cells in a transgenic dioxin-responsive lacZ mouse model after acute TCDD exposure. AhR and Arnt were also expressed in cerebellar granule neuroblast cultures. AhR localized to the nucleus in granule cells 15 min after TCDD treatment. TCCD elicited time-dependent and concentration-dependent increases in CYP1A1 and 1B1 mRNA and protein levels. Moreover, TCDD treatment reduced both thymidine incorporation and granule neuroblast survival in a concentration-dependent manner. These data suggest that (1) granule neuroblasts are direct targets for developmental AhR-mediated TCDD neurotoxicity and (2) TCDD exposure may disrupt granule cell neurogenesis. PMID- 15537748 TI - Induction of cell-cycle arrest by all-trans retinoic acid in mouse embryonic palatal mesenchymal (MEPM) cells. AB - all-trans retinoic acid (atRA), the oxidative metabolite of vitamin A, is essential for normal embryonic development. Also, high levels of atRA are teratogenic in many species and can effectively induce cleft palate in the mouse. Most cleft palate resulted from the failed fusion of secondary palate shelves, and maintenance of the normal cell proliferation is important in this process of shelf growth. To clarify the mechanism by which atRA causes cleft palate, we investigated the effect of atRA on proliferation activity and cell cycle distribution in mouse embryonic palatal mesenchymal (MEPM) cells. atRA inhibited the growth of MEPM cells by inducing apoptosis in a dose-dependent manner. atRA also caused a G1 block in the cell cycle with an increase in the proportion of cells in G0/G1 and a decrease in the proportion of cells in S phase, as determined by flow cytometry. We next investigated the effects of atRA on molecules that regulate the G1 to S phase transition. These studies demonstrated that atRA inhibited expression of cyclins D and E at the protein level. Furthermore, atRA treatment reduced phosphorylated Rb and decreased cdk2 and cdk4 kinase activity. These data suggest that atRA had antiproliferative activity by modulating G1/S cell cycle regulators and by inhibition of Rb phosphorylation in MEPM cells, which might account for the pathogenesis of cleft palate induced by retinoic acid. PMID- 15537749 TI - Aluminum-maltolate induces apoptosis and necrosis in neuro-2a cells: potential role for p53 signaling. AB - Aluminum maltolate (Al-malt) causes neurodegeneration following in vivo exposure, and apoptosis plays a prominent role. The objective of this study was to define the form of cell death induced by Al-malt and to establish an in vitro model system amenable to mechanistic investigations of Al-malt-induced cell death. Neuro-2a cells, a murine neuroblastoma cell line, were treated with Al-malt for 24 h, following which mode of cell death and alterations in apoptosis-related gene expression were studied. Al-malt concentration-dependently increased cell death. The mode of cell death was a combination of apoptosis and necrosis. Treatment with Al-malt resulted in caspase 3 activation and the externalization of phosphatidyl serine, both indicative of apoptosis. In addition, nuclear condensation and fragmentation were evident. Interestingly, pretreatment with cycloheximide (CHX), a potent protein synthesis inhibitor markedly reduced Al malt-induced apoptosis, indicating that altered gene expression was critical for this form of cell death. Pretreatment with CHX had no effect on necrosis induced by Al-malt. Analysis of gene expression showed that p53 mRNA was increased following treatment with Al-malt. This increase was accompanied by a marked inhibition of Bcl2 expression and an increase in BAX expression, a pattern of gene expression suggestive of a pro-apoptotic shift. Results show for the first time that p53 is induced by Al in neuron-like cells and suggest that the p53 dependent intrinsic pathway may be responsible for Al-induced apoptosis. Future studies investigating the role of p53 in Al neurotoxicity both in vivo and in vitro are warranted. PMID- 15537750 TI - FTIR reveals structural differences between native beta-sheet proteins and amyloid fibrils. AB - The presence of beta-sheets in the core of amyloid fibrils raised questions as to whether or not beta-sheet-containing proteins, such as transthyretin, are predisposed to form such fibrils. However, we show here that the molecular structure of amyloid fibrils differs more generally from the beta-sheets in native proteins. This difference is evident from the amide I region of the infrared spectrum and relates to the distribution of the phi/psi dihedral angles within the Ramachandran plot, the average number of strands per sheet, and possibly, the beta-sheet twist. These data imply that amyloid fibril formation from native beta-sheet proteins can involve a substantial structural reorganization. PMID- 15537751 TI - Possible role of region 152-156 in the structural duality of a peptide fragment from sheep prion protein. AB - The conformational conversion of the nonpathogenic "cellular" prion isoform into a pathogenic "scrapie" protease-resistant isoform is a fundamental event in the onset of transmissible spongiform encephalopathies (TSE). During this pathogenic conversion, helix H1 and its two flanking loops of the normal prion protein are thought to undergo a conformational transition into a beta-like structure. A peptide spanning helix H1 and beta-strand S2 (residues 142-166 in human numbering) was studied by circular dichroism and nuclear magnetic resonance spectroscopies. This peptide in aqueous solution, in contrast to many prion fragments studied earlier (1) is highly soluble and (2) does not aggregate until the millimolar concentration range, and (3) exhibits an intrinsic propensity to a beta-hairpin-like conformation at neutral pH. We found that this peptide can also fold into a helix H1 conformation when dissolved in a TFE/PB mixture. The structures of the peptide calculated by MD showed solvent-dependent internal stabilizing forces of the structures and evidenced a higher mobility of the residues following the end of helix H1. These data suggest that the molecular rearrangement of this peptide in region 152-156, particularly in position 155, could be associated with the pathogenic conversion of the prion protein. PMID- 15537752 TI - Interaction of the N-terminal domain of Escherichia coli heat-shock protein ClpB and protein aggregates during chaperone activity. AB - The Escherichia coli heat-shock protein ClpB reactivates protein aggregates in cooperation with the DnaK chaperone system. The ClpB N-terminal domain plays an important role in the chaperone activity, but its mechanism remains unknown. In this study, we investigated the effect of the ClpB N-terminal domain on malate dehydrogenase (MDH) refolding. ClpB reduced the yield of MDH refolding by a strong interaction with the intermediate. However, the refolding kinetics was not affected by deletion of the ClpB N-terminal domain (ClpBDeltaN), indicating that MDH refolding was affected by interaction with the N-terminal domain. In addition, the MDH refolding yield increased 50% in the presence of the ClpB N terminal fragment (ClpBN). Fluorescence polarization analysis showed that this chaperone-like activity is explained best by a weak interaction between ClpBN and the reversible aggregate of MDH. The dissociation constant of ClpBN and the reversible aggregate was estimated as 45 muM from the calculation of the refolding kinetics. Amino acid substitutions at Leu 97 and Leu 110 on the ClpBN surface reduced the chaperone-like activity and the affinity to the substrate. In addition, these residues are involved in stimulation of ATPase activity in ClpB. Thus, Leu 97 and Leu 110 are responsible for the substrate recognition and the regulation of ATP-induced ClpB conformational change. PMID- 15537753 TI - Native and nonnative conformational preferences in the urea-unfolded state of barstar. AB - The refolding of barstar from its urea-unfolded state has been studied extensively using various spectroscopic probes and real-time NMR, which provide global and residue-specific information, respectively, about the folding process. Here, a preliminary structural characterization by NMR of barstar in 8 M urea has been carried out at pH 6.5 and 25 degrees C. Complete backbone resonance assignments of the urea-unfolded protein were obtained using the recently developed three-dimensional NMR techniques of HNN and HN(C)N. The conformational propensities of the polypeptide backbone in the presence of 8 M urea have been estimated by examining deviations of secondary chemical shifts from random coil values. For some residues that belong to helices in native barstar, 13C(alpha) and 13CO secondary shifts show positive deviations in the urea-unfolded state, indicating that these residues have propensities toward helical conformations. These residues are, however, juxtaposed by residues that display negative deviations indicative of propensities toward extended conformations. Thus, segments that are helical in native barstar are unlikely to preferentially populate the helical conformation in the unfolded state. Similarly, residues belonging to beta-strands 1 and 2 of native barstar do not appear to show any conformational preferences in the unfolded state. On the other hand, residues belonging to the beta-strand 3 segment show weak nonnative helical conformational preferences in the unfolded state, indicating that this segment may possess a weak preference for populating a helical conformation in the unfolded state. PMID- 15537754 TI - Calcium(II) selectively induces alpha-synuclein annular oligomers via interaction with the C-terminal domain. AB - Alpha-synuclein filaments are the major component of intracytoplasmic inclusion bodies characteristic of Parkinson's disease and related disorders. The process of alpha-synuclein filament formation proceeds via intermediate or protofibrillar species, each of which may be cytotoxic. Because high levels of calcium(II) and other metal ions may play a role in disease pathogenesis, we investigated the influence of calcium and other metals on alpha-synuclein speciation. Here we report that calcium(II) and cobalt(II) selectively induce the rapid formation of discrete annular alpha-synuclein oligomeric species. We used atomic force microscopy to monitor the aggregation state of alpha-synuclein after 1 d at 4 degrees C in the presence of a range of metal ions compared with the filament formation pathway in the absence of metal ions. Three classes of effect were observed with different groups of metal ions: (1) Copper(II), iron(III), and nickel(II) yielded 0.8-4 nm spherical particles, similar to alpha-synuclein incubated without metal ions; (2) magnesium(II), cadmium(II), and zinc(II) gave larger, 5-8 nm spherical oligomers; and, (3) cobalt(II) and calcium(II) gave frequent annular oligomers, 70-90 nm in diameter with calcium(II) and 22-30 nm in diameter with cobalt(II). In the absence of metal ions, annular oligomers ranging 45-90 nm in diameter were observed after 10 d incubation, short branched structures appeared after a further 3 wk and extended filaments after 2-3 mo. Previous studies have shown that alpha-synuclein calcium binding is mediated by the acidic C terminus. We found that truncated alpha-synuclein (1-125), lacking the C-terminal 15 amino acids, did not form annular oligomers upon calcium addition, indicating the involvement of the calcium-binding domain. PMID- 15537755 TI - Structure of human brain fructose 1,6-(bis)phosphate aldolase: linking isozyme structure with function. AB - Fructose-1,6-(bis)phosphate aldolase is a ubiquitous enzyme that catalyzes the reversible aldol cleavage of fructose-1,6-(bis)phosphate and fructose 1-phosphate to dihydroxyacetone phosphate and either glyceral-dehyde-3-phosphate or glyceraldehyde, respectively. Vertebrate aldolases exist as three isozymes with different tissue distributions and kinetics: aldolase A (muscle and red blood cell), aldolase B (liver, kidney, and small intestine), and aldolase C (brain and neuronal tissue). The structures of human aldolases A and B are known and herein we report the first structure of the human aldolase C, solved by X-ray crystallography at 3.0 A resolution. Structural differences between the isozymes were expected to account for isozyme-specific activity. However, the structures of isozymes A, B, and C are the same in their overall fold and active site structure. The subtle changes observed in active site residues Arg42, Lys146, and Arg303 are insufficient to completely account for the tissue-specific isozymic differences. Consequently, the structural analysis has been extended to the isozyme-specific residues (ISRs), those residues conserved among paralogs. A complete analysis of the ISRs in the context of this structure demonstrates that in several cases an amino acid residue that is conserved among aldolase C orthologs prevents an interaction that occurs in paralogs. In addition, the structure confirms the clustering of ISRs into discrete patches on the surface and reveals the existence in aldolase C of a patch of electronegative residues localized near the C terminus. Together, these structural changes highlight the differences required for the tissue and kinetic specificity among aldolase isozymes. PMID- 15537756 TI - Analysis of side chain mobility among protein G B1 domain mutants with widely varying stabilities. AB - "Host-guest" studies of the B1 domain from Streptococcal protein G have been used previously to establish a thermodynamic scale for the beta-sheet-forming propensities of the 20 common amino acids. To investigate the contribution of side chain conformational entropy to the relative stabilities of B1 domain mutants, we have determined the dynamics of side chain methyl groups in 10 of the 20 mutants used in a previous study. Deuterium relaxation rates were measured using two-dimensional NMR techniques for 13CH2D groups. Analysis of the relaxation data using the Lipari-Szabo model-free formalism showed that mutations introduced at the guest position caused small but statistically significant changes in the methyl group dynamics. In addition, there was a low level of covariation of the Lipari-Szabo order parameters among the 10 mutants. The variations in conformational free energy estimated from the order parameters were comparable in magnitude to the variations in global stability of the 10 mutants but did not correlate with the global stability of the domain or with the structural properties of the guest amino acids. The data support the view that conformational entropy in the folded state is one of many factors that can influence the folding thermodynamics of proteins. PMID- 15537757 TI - Modulation of the ligand binding properties of the transcription repressor NmrA by GATA-containing DNA and site-directed mutagenesis. AB - NmrA is a negative transcription-regulating protein that binds to the C-terminal region of the GATA transcription-activating protein AreA. The proposed molecular mechanism of action for NmrA is to inhibit AreA binding to its target promoters. In contrast to this proposal, we report that a C-terminal fragment of AreA can bind individually to GATA-containing DNA and NmrA and that in the presence of a mixture of GATA-containing DNA and NmrA, the AreA fragment binds preferentially to the GATA-containing DNA in vitro. These observations are consistent with NmrA acting by an indirect route, such as by controlling entry into the nucleus. Deletion of the final nine amino acids of a C-terminal fragment of AreA does not affect NmrA binding. Wild-type NmrA binds NAD(+)(P+) with much greater affinity than NAD(P)H, despite the lack of the consensus GXXGXXG dinucleotide-binding motif. However, introducing the GXXGXXG sequence into the NmrA double mutant N12G/A18G causes an approximately 13-fold increase in the KD for NAD+ and a 2.3 fold increase for NADP+. An H37W mutant in NmrA designed to increase the interaction with the adenine ring of NAD+ has a decrease in KD of approximately 4.5-fold for NAD+ and a marginal 24% increase for NADP+. The crystal structure of the N12G/A18G mutant protein shows changes in main chain position as well as repositioning of H37, which disrupts contacts with the adenine ring of NAD+, changes which are predicted to reduce the binding affinity for this dinucleotide. The substitutions E193Q/D195N or Q202E/F204Y in the C-terminal domain of NmrA reduced the affinity for a C-terminal fragment of AreA, implying that this region of the protein interacts with AreA. PMID- 15537758 TI - Identification and fine mapping of quantitative trait loci for growth traits on bovine chromosomes 2, 6, 14, 19, 21, and 23 within one commercial line of Bos taurus. AB - We report the identification and fine mapping of QTL for birth weight (BWT), preweaning ADG (PWADG), and postweaning ADG on feed (ADGF) in a commercial line of Bos taurus using an identical-by-descent haplotype sharing method. One hundred seventy-six calves of 12 bulls (9 to 30 male calves from each sire) of the Beefbooster, Inc., M1 line were typed using 71 genetic markers from bovine chromosomes (BTA) 2, 6, 14, 19, 21, and 23 (8 to 16 markers from each chromosome). Sixteen haplotypes were found to have significant (P <0.05) associations with BWT at the comparison-wise threshold. The 16 haplotypes span 13 chromosomal regions, two on BTA 2 (9.1 to 22.5 cM and 95.0 to 100.3 cM), three on BTA 6 (8.2 to 11.8 cM, 35.5 to 49.7 cM, and 83.0 to 86.2 cM), three on BTA 14 (26.0 to 26.7 cM, 36.2 to 46.2 cM, and 52.0 to 67.7 cM), one on BTA 19 (52.0 to 52.7 cM), two on BTA 21 (9.9 to 20.4 cM and 28.2 to 46.1 cM), and two on BTA 23 (23.9 to 36.0 cM and 45.1 to 50.9 cM). Thirteen haplotypes spanning seven chromosomal regions significantly affected (P <0.05) PWADG at the comparison-wise threshold. The seven chromosomal regions include two regions on BTA 6 (11.8 to 44.2 cM and 83.0 to 86.2 cM), one on BTA 14 (26.7 to 50.8 cM), one on BTA 19 (4.8 to 15.9 cM), one on BTA 21 (9.9 to 20.4 cM), and two on BTA 23 (17.3 to 36.0 cM and 45.1 to 50.9 cM). For ADGF, 11 haplotypes were identified to have significant associations (P <0.05) at the comparison-wise threshold. The 11 haplotypes represented eight chromosomal regions, one on BTA 2 (9.1 to 22.5 cM), two on BTA 6 (49.7 to 50.1 cM and 59.6 to 63.6 cM), two on BTA 14 (17.0 to 24.0 cM and 36.2 to 46.2 cM), two on BTA 19 (52.0 to 52.7 cM and 65.1 to 65.7 cM), and one on BTA 21 (46.1 to 53.1 cM). The QTL regions identified and fine mapped in this study will provide a reference for future positional candidate gene research and marker assisted selection of various growth traits. PMID- 15537759 TI - Quantitative trait loci analysis for growth and carcass traits in a half-sib family of purebred Japanese Black (Wagyu) cattle. AB - We used a half-sib family of purebred Japanese Black (Wagyu) cattle to locate economically important quantitative trait loci. The family was composed of 348 fattened steers, 236 of which were genotyped for 342 microsatellite markers spanning 2,664 cM of 29 bovine autosomes. The genome scan revealed evidence of 15 significant QTL (<5% chromosome-wise level) affecting growth and carcass traits. Of the 15 QTL, six QTL were significant at the 5% experiment-wise level and were located in bovine chromosomes (BTA) 4, 5, and 14. We analyzed these three chromosomes in more detail in the 348 steers, with an average marker interval of 1.2 cM. The second scan revealed that the same haplotype of the BTA 4 region (52 to 67 cM) positively affected LM area and marbling. We confirmed the QTL for carcass yield estimate on BTA 5 in the region of 45 to 54 cM. Five growth-related QTL located on BTA 14, including slaughter and carcass weights, were positively affected by the same region of the haplotype of BTA 14 (29-51 cM). These data should provide a useful reference for further marker-assisted selection in the family and positional cloning research. The research indicates that progeny design with moderate genotyping efforts is a powerful method for detecting QTL in a purebred half-sib family. PMID- 15537760 TI - A genome scan for quantitative trait loci and imprinted regions affecting reproduction in pigs. AB - Quantitative trait loci for reproductive traits in a three-generation resource population of a cross between low-indexing pigs from a control line and high indexing pigs from a line selected 10 generations for increased index of ovulation rate and embryonic survival are reported. Phenotypic data were collected in F2 females for birth weight (BWT, n = 428), weaning weight (WWT, n = 405), age at puberty (AP, n = 295), ovulation rate (OR, n = 423), number of fully formed pigs (FF, n = 370), number of pigs born alive (NBA, n = 370), number of mummified pigs (MUM, n = 370), and number of stillborn pigs (NSB, n = 370). Grandparent, F1, and F2 animals were genotyped for 151 microsatellite markers. Sixteen putative QTL (P < 0.10) for reproductive traits were identified in previous analyses of these data with single QTL line-cross models. Data were reanalyzed with multiple QTL models, including imprinting effects. Data also were analyzed with half-sib models. Permutation was used to establish genome-wide significance levels ( = 0.01, 0.05, and 0.10). Thirty-one putative QTL for reproductive traits and two QTL for birth weight were identified (P < 0.10). One Mendelian QTL for FF (P < 0.05), one for NBA (P < 0.05), three for NSB (P < 0.05), three for NN (P < 0.05), seven for AP (P < 0.10), five for MUM (P < 0.10), and one for BWT (P < 0.10) were found. Partial imprinting of QTL affecting OR (P < 0.01), BWT (P < 0.05), and MUM (P < 0.05) was detected. There were four paternally expressed QTL for NN (P < 0.10) and one each for AP (P < 0.05) and MUM (P < 0.10). Maternally expressed QTL affecting NSB (P < 0.10), NN (P < 0.10), and MUM (P < 0.10) were detected. No QTL were detected with half-sib analyses. Multiple QTL models with imprinting effects are more appropriate for analyzing F2 data than single Mendelian QTL line-cross models. PMID- 15537761 TI - Joint analysis of multiple cDNA microarray studies via multivariate mixed models applied to genetic improvement of beef cattle. AB - In functional genomic laboratories, it is common to use the same microarray slide across studies, each investigating a unique biological question, and each analyzed separately due to computational limitations and/or because there is no hybridization of samples from different studies on one slide. However, the question of analyzing data from multiple studies is a major current issue in microarray data analysis because there are gains to be made in the accuracy of estimated effects by exploiting a covariance structure between gene expression data across studies. We propose an approach for combining multiple studies using multivariate mixed models, with the assumption of a nonzero correlation among genes across experiments, while imposing a null residual covariance. We applied this method to jointly analyze three experiments in genetics of cattle with a total of 54 arrays, each with 19,200 spots and 7,638 elements. The resulting seven-variate model contains 752,476 equations and 56 covariances. To identify differentially expressed genes, we applied model-based clustering to a linear combination of the random gene x variety interaction effect. We enhanced the biological interpretation of the results by applying an iterative algorithm to identify the gene ontology classes that significantly changed in each experiment. We found 118 elements with coordinate expression that clustered into distinct biological functions such as adipogenesis and protein turnover. These results contribute to our understanding of the mechanistic processes involved in adipogenesis and nutrient partitioning. PMID- 15537762 TI - Genetic relationships between calving performance and beef production traits in Piemontese cattle. AB - The aim of the study was to obtain estimates of genetic correlations between direct and maternal calving performance of heifers and cows and beef production traits in Piemontese cattle. Beef production traits were daily gain, live fleshiness, and bone thinness measured on 1,602 young bulls tested at a central station. Live fleshiness (six traits) and bone thinness were subjectively scored by classifiers using a nine-point linear grid. Data on calving performance were calving difficulty scores (five classes from unassisted to embryotomy) routinely recorded in the farms. Calving performance of heifers and cows were considered different traits. A total of 30,763 and 80,474 calving scores in first and later parities, respectively, were used to estimate covariance components with beef traits. Data were analyzed using bivariate linear animal models, including direct genetic effects for calving performance and beef traits and maternal genetic effects only for calving performance. Due to the nature of the data structure, which involved traits measured in different environments and on different animals, covariances were estimated mostly through pedigree information. Genetic correlations of daily gain were positive with direct calving performance (0.43 in heifers and 0.50 in cows) and negative with maternal calving performance (-0.23 and -0.28 for heifers and cows, respectively). Live fleshiness traits were moderately correlated with maternal calving performance in both parities, ranging from 0.06 to 0.33. Correlations between live fleshiness traits and direct calving performance were low to moderate and positive in the first parity, but trivial in later parities. Bone thinness was negatively correlated with direct calving performance (-0.17 and -0.38 in heifers and cows, respectively), but it was positively correlated to maternal calving performance (0.31 and 0.40). Estimated residual correlations were close to zero. Results indicate that, due to the existence of antagonistic relationships between the investigated traits, specific selection strategies need to be studied. PMID- 15537763 TI - Comparing alternative definitions of the contemporary group effect in Avilena Negra Iberica beef cattle using classical and Bayesian criteria. AB - Data on weaning weight from 12,740 animals were used to compare different definitions of contemporary groups (CG) for the genetic evaluation of the Avilena Negra Iberica beef cattle breed. Six alternative definitions for the CG effect were considered: herd-year-season of calving (HYS), with seasons defined according to the four natural seasons; herd-year-month of calving (HYM); herd clusters of 30 d (HC30-30) or 90 d (HC90-90); and adaptive herd clusters with two time limits, 30 and 90 d (HC30-90), and 30 and 180 d (HC30-180). A minimum of five observations in each CG class was required. This rendered substantial differences in loss of information, ranging from 0.7% of the total number of records for HC30-180 to 14% for HYM. Several classical statistics and Bayesian criteria for statistical model comparison were used. The use of classical criteria, such as the between- and within-CG variation and the accuracy of prediction, can be controversial because of their dependency on the unknown variance components. Residual variance decreased with the decrease in time span associated with the definition of CG. This was expected in this population because environmental conditions are highly variable throughout the year. However, estimates of the additive genetic variance for direct effects, which should not be affected by the definition of CG, were substantially larger for definitions involving larger time periods (HYS, HC90-90). When parameters used in the current evaluation procedure were used with all data sets, CG involving 30 d (HYM and HC30-30) were optimal in terms of providing the lowest/largest within /between-CG variation. On the other hand, CG involving 90 d (HYS and HC90-90) yielded the poorest within-/between CG variation, with only a slight improvement of accuracy of prediction of direct genetic values over the other definitions. Bayes factors and cross-validation predictive densities allowed for improved discrimination among models. Models including CG spanning 30 d were more plausible and showed better predicting ability than models spanning 90 d. Adaptive CG showed intermediate results. Overall, it seems that average time span rendered by the different definitions had a major effect on the ranking of models. However, from the breeder's point of view, the loss of information associated with definitions involving shorter periods of time, such as HYM or HC30-30, might be unacceptable. PMID- 15537764 TI - Genetic correlations between reproduction and production traits in swine. AB - Genetic correlations between reproduction and production traits were estimated in swine. Reproduction traits investigated were age at first service (AFS), number of live-born piglets in the first litter (NBA1), interval from weaning to first service after first litter (WTS1), number of live-born piglets in the second litter (NBA2), and interval from weaning to first service after the second litter (WTS2). Females generating the data were Norwegian Landrace born in nucleus herds between 1990 and 2000, and the number of records ranged from 13,792 to 56,932. Genetic correlations were estimated among the main production traits in the breeding goal: adjusted age at 100 kg live weight (A100), percentage of lean meat content (LMC), individual feed consumption from 25 to 100 kg (FC), and bacon side quality (BSQ). Average adjusted backfat thickness (BF) was included as a production trait. The A100 and BF traits were recorded on gilts on-farm with 190,454 records, whereas LMC, BSQ, and FC were recorded on-station with the number of records ranging from 12,487 to 12,992. Analyses were carried out with a multivariate animal model using average information restricted maximum likelihood procedures by first running each reproduction trait with A100 and BF, followed by each reproduction trait with LMC, BSQ, and FC. Average heritabilities for reproduction traits were as follows: AFS (0.38), NBA1 (0.11), WTS1 (0.06), NBA2 (0.12), and WTS2 (0.03); and for production traits: A100 (0.30), BF (0.44), FC (0.22), LMC (0.58), and BSQ (0.23). The highest genetic correlation was estimated between A100 and AFS (r(g)= 0.68), also resulting in a positive genetic correlation between FC and AFS. Growth (A100) was negatively (i.e., unfavorably) genetically correlated to NBA1 and NBA2 (r(g) = 0.60 and rg = 0.42 respectively), and so the genetic correlation to FC also became unfavorable (r(g)= 0.23 and r(g) = 0.20). Single-trait selection for enhanced LMC would also affect NBA1 and NBA2 unfavorably (r(g)= -0.12 and r(g)= -0.24). Correlations between BF at 100 kg live weight and reproduction traits were close to zero; however, a low genetic correlation between BF and WTS1 was obtained (r(g)= -0.12), indicating that selection toward reduced BF at 100 kg live weight may have an unfavorable impact on WTS1. PMID- 15537765 TI - Use of structured antedependence models for the genetic analysis of growth curves. AB - Growth curve analysis is an important issue for many agricultural and laboratory species, for both phenotypic and genetic studies. The aim of this paper is to present the use of a novel statistical approach, namely the structured antedependence (SAD) models, to deal with this issue. The basic idea of these models is that an observation at time t can be explained by the previous observations. These models are especially appropriate to deal with cumulative traits such as growth, as BW at age t clearly depends on BW measures at ages (t 1), (t -2), etc. These models were applied on an INRA experimental Charolais herd data set. The data comprised BW records for 560 cows born over an 11-yr period (from 1988 to 1998) from 60 sires and 369 dams. The proposed SAD models were compared with the well-known random regression (RR) models that are already widely used in various areas of longitudinal data analysis. It was found that the SAD models fit the growth process better with far fewer parameters than the RR models (9 instead of 16 covariance parameters for the phenotypic analysis, and 14 instead of 21 for the genetic analysis). Despite this smaller number of covariance parameters, the likelihood value was found to be much higher with the SAD vs. the RR models, with a difference of 262.9 for the phenotypic analysis with a quartic polynomial for the RR and 751.5 for the genetic analysis with a cubic polynomial for both the genetic and environmental parts of the RR model. The SAD models also proved to be better able to interpolate missing values. Heritability, genetic, and environmental correlation coefficients were estimated for weights from birth to adulthood. The structured antedependence models proved, in this study, to be very appropriate to model growth data in a parsimonious and flexible way. PMID- 15537766 TI - Association of markers in the bovine CAPN1 gene with meat tenderness in large crossbred populations that sample influential industry sires. AB - Two previously identified single-nucleotide polymorphism markers located within the micromolar calcium-activated neutral protease gene (CAPN1) were evaluated for their association with variation in meat tenderness using one commercial sample of Simmental x Angus crossbred calves and one multibreed, crossbred research herd. The commercial sample included 362 animals sired by 23 registered Simmental bulls bred to unregistered Angus cows and represented current industry animals in which to test the predictive merit of the markers. The second sample was a research herd including 564 steers from the Germplasm Evaluation Cycle VII population at the U.S. Meat Animal Research Center, produced with semen from popular sires of the seven Bos taurus beef breeds with the most registrations in the United States (Angus, Charolais, Gelbvieh, Hereford, Limousin, Red Angus, and Simmental) on Angus, Hereford, and MARC III cows. These animals form a relatively outbred population that constituted a stringent test of the predictive merit of the genetic markers, although small groups were half-sibs. Warner-Bratzler shear force measurements were used to determine tenderness phenotypes for all animals. The populations were genotyped for two markers that predict variation at amino acid positions 316 and 530 of the mu-calpain polypeptide, produced by the CAPN1 gene. Minor allele frequencies for markers 316 and 530 in the commercial sample were 0.17 and 0.37, respectively, and in the Cycle VII animals, were 0.20 and 0.28, respectively. Both markers showed association with shear force in the commercial sample (P = 0.04) and the Cycle VII population (P = 0.02), supporting the hypothesis that they represent potential markers to aid selection for improved meat tenderness in commercial populations of beef cattle in the United States. PMID- 15537767 TI - National Pork Producers Council Maternal Line Genetic Evaluation: a comparison of growth and carcass traits in terminal progeny. AB - The objective of this study was to compare growth and carcass traits of 1,252 progeny of six commercially available dam lines included in the National Pork Producers Council Maternal Line Evaluation Project. Lines compared included one maternal line supplied by each of American Diamond Swine Genetics (ADSG), Danbred NA (DB), two lines supplied by Monsanto Choice Genetics (DK and GPK347), Newsham Hybrids (NH), and Landrace x Large White females supplied by the National Swine Registry (NSR). All females were mated to DB, Duroc-Hampshire terminal sires. Traits analyzed were ADG from 56 to 115 kg live weight, days to 115 kg, backfat thickness measured at the 10th rib, carcass length, dressing percent, and 10th rib LM area. Carcass traits were adjusted to a carcass weight of 85 kg. The statistical model included fixed effects of maternal line, sex, farrowing group, and finishing unit (farm). All two-way interactions among main effects were tested and removed from final models because they were not significant. In addition, because they were not significant, effects of farm and farrowing group were removed from models for carcass length and 10th-rib backfat thickness, and farm was removed from the model for LM area. Least squares means for ADG ranged from 0.74 to 0.79 kg/d. The GPK347 line had lower ADG and greater days to 115 kg than all other lines (P < 0.05). The ADSG (P < 0.05) and NH (P < 0.01) progeny had lower ADG than DK progeny. The DK line had the fewest days to 115 kg (P < 0.05). Progeny for the DB and NH lines had the least 10th-rib backfat, differing from ADSG, DK, and GPK347 (P < 0.05). Pigs from DB females had the greatest dressing percent, differing from ADSG, DK, GPK347, and NH (P < 0.05). The GPK347 had a lower dressing percent than all other lines (P < 0.05). Progeny of DB females had the greatest LM area, differing from ADSG, DK, GPK347, and NSR (P < 0.05). Offspring from ADSG and GPK347 had the smallest LM area; however, GPK347 and NSR did not differ. Differences in carcass length were statistically significant; however, actual differences were small. Economic weights for these traits relative to reproductive traits must be considered in integrated economic analyses to properly compare differences among lines in net economic value for specific markets. PMID- 15537768 TI - Hormonal regulation of leptin and leptin receptor expression in porcine subcutaneous adipose tissue. AB - The current study was performed to examine the response of the leptin gene to hormonal stimuli in porcine adipose tissue from finishing pigs. Yorkshire gilts (approximately 150 kg BW) were used in this study. Tissue from four to six pigs was used per experiment. Dorsal subcutaneous adipose tissue samples were acquired, and adipose tissue explants (approximately 100 mg) were prepared using sterile technique. Tissue slices were transferred to 12-well tissue culture plates containing 1 mL of Media 199 with 25 mM HEPES, 0.5% BSA, pH 7.4, and various hormone supplements. Triplicate tissue slices were incubated with either basal medium or hormone-supplemented media in a tissue culture incubator at 37 degrees C with 95% air:5% CO2. Hormones included insulin (100 nM), dexamethasone (1 microM), porcine GH, 100 ng/mL), triiodothyronine (T3, 10 nM), porcine leptin (100 ng/mL), or IGF-I (250 ng/mL). Following incubation for 24 h, tissue samples from the incubations were blotted and transferred to microfuge tubes, frozen in liquid N, and stored at -80 degrees C before analysis for gene mRNA abundance by reverse-transcription PCR and subsequent quantification of transcripts by capillary electrophoresis with laser-induced fluorescence detection. Media from the incubations were collected in microfuge vials and stored at -20 degrees C before analysis for leptin content by RIA. Insulin was required to maintain tissue and mRNA integrity; therefore, insulin was included in all incubations. The combination of insulin and dexamethasone stimulated leptin secretion into the medium by 60% (P < 0.05; n = 6). Porcine GH inhibited insulin induced leptin secretion by 25% (P < 0.05; n = 6). Dexamethasone in combination with insulin produced a 22% increase in leptin mRNA abundance relative to insulin (P < 0.05; n = 4), and T3 stimulated a 28% increase in insulin-induced leptin mRNA abundance (P < 0.05; n = 4). Leptin receptor mRNA abundance was decreased by 25% with the combination of insulin and dexamethasone, relative to insulin-treated adipose tissue slices (P < 0.05; n = 4). Porcine GH decreased leptin receptor mRNA abundance by 17% (P < 0.05; n = 6). These data suggest that leptin secretion is a regulated phenomenon and that posttranslational processing may be significant. Alternatively, transport and exocytosis of leptin containing vesicles in the pig adipocyte may be quite complicated, which could account for the differences in observed mRNA abundance and protein secretion. PMID- 15537769 TI - Effect of heat exposure on uncoupling protein-3 mRNA abundance in porcine skeletal muscle. AB - Exposure to cold increases abundance of mRNA for uncoupling protein-3 (UCP3) in skeletal muscle, whereas the influence of exposure to heat is unknown. Thus, we conducted a study to investigate the influence of heat exposure on UCP3 mRNA abundance in porcine skeletal muscle. Three pigs aged 110 to 120 d, with an average BW of 75 kg, from each of eight litters were used. Each littermate was assigned to one of three treatment groups; one group was reared at 32 degrees C and fed ad libitum (32AL) for 4 wk, whereas the other two groups were maintained at 23 degrees C for the same period, and either pair-fed the intake of their 32AL littermates (23PF), or fed ad libitum (23AL). The RNase protection assay revealed that UCP3 mRNA abundance in longissimus dorsi and rhomboideus muscles was higher (P < 0.05) in the 32AL group than the 23PF group. The 23AL group also had significantly higher UCP3 mRNA abundance than the 23PF group in these muscles. Plasma total 3,5,3'-triiodothyronine concentration of the 32AL group was lower (P < 0.05) than that of the 23PF group, whereas mRNA abundance of thyroid hormone receptor (TR) isoforms, TRalpha1 and TRalpha2, in these muscles was not affected, suggesting that the 32AL group was in a relatively hypo-thyroid state. Because thyroid hormone up-regulates UCP3 expression, these results indicate that factors other than thyroid hormone may play a role in regulating UCP3 mRNA abundance in skeletal muscle of heat-exposed pigs. PMID- 15537770 TI - Effect of chronic clenbuterol administration and exercise training on immune function in horses. AB - Effects of longitudinal exercise training and acute intensive exercise (simulated race test) on immune function have not been reported in horses. Clenbuterol, a beta2-adrenergic agonist, is used to manage inflammatory airway disease in horses. This study investigated the interaction of 8 wk of exercise training with or without 12 wk of clenbuterol administration in horses. Twenty-three untrained standardbred mares (10 +/- 3 yr, Mean +/- SE) were used and divided into four experimental groups. Horses given clenbuterol plus exercise (CLENEX; n = 6) and clenbuterol alone (CLEN; n = 6) received 2.4 microg/kg BW of clenbuterol twice daily (in an average volume of 20 mL) on a schedule of 5 d on and 2 d off for 12 wk. The CLENEX group was also aerobically trained 3 d/wk. Mares given exercise alone (EX; n = 5) were aerobically trained for 3 d/wk, and the control group (CON; n = 6) remained sedentary. Both EX and CON horses were administered similar volumes (approximately 20 mL) of molasses twice daily. A simulated race test (SRT) resulted in an elevation in lymphocyte number postexercise (P < 0.05). There was no significant difference after acute exercise in either monocyte or granulocyte number. Acute exercise resulted in a decrease (P < 0.05) in the percentage of CD4+ and an increase (P < 0.05) in the percentage of CD8+ cells. The SRT resulted in a decreased lymphoproliferative response to pokeweed mitogen (P < 0.05). A SRT had no effect on antibody production in response to equine influenza vaccine. The EX group demonstrated greater cortisol concentrations at rest and at all other time points postexercise after completing the training regimen compared with CLENEX horses (P < 0.05). Preexercise (SRT) peripheral blood monocyte number was lower in CLENEX horses than in other treatment groups (P < 0.05). Clenbuterol and exercise training did not significantly affect post SRT changes in leukocyte numbers. Exercise training resulted in a decrease (P < 0.05) in the percentage of CD8+ cells post-SRT compared with other groups, but the percentage of CD4+ cells was not altered by either clenbuterol or exercise conditioning. Lymphocyte proliferative response was not affected by clenbuterol or exercise treatment. Horses demonstrated responses to bouts of acute exercise as noted with other species, namely humans and rodents. PMID- 15537771 TI - Effect of addition of a progesterone intravaginal insert to a timed insemination protocol using estradiol cypionate on ovulation rate, pregnancy rate, and late embryonic loss in lactating dairy cows. AB - The objectives of this study were to determine the effects of incorporating a progesterone intravaginal insert (CIDR) between the day of GnRH and PGF2alpha treatments of a timed AI protocol using estradiol cypionate (ECP) to synchronize ovulation on display of estrus, ovulation rate, pregnancy rate, and late embryonic loss in lactating cows. Holstein cows, 227 from Site 1 and 458 from Site 2, were presynchronized with two injections of PGF2alpha on study d 0 and 14, and subjected to a timed AI protocol (100 mixrog of GnRH on study d 28, 25 mg of PGF2alpha on study d 35, 1 mg of ECP on study d 36, and timed AI on study d 38) with or without a CIDR insert. Blood was collected on study d 14 and 28 for progesterone measurements to determine cyclicity. Ovaries were scanned on d 35, 37, and 42, and pregnancy diagnosed on d 65 and 79, which corresponded to 27 and 41 d after AI. Cows receiving a CIDR had similar rates of detected estrus (77.2 vs. 73.8%), ovulation (85.6 vs. 86.6%), and pregnancy at 27 (35.8 vs. 38.8%) and 41 d (29.3 vs. 32.3%) after AI, and late embryonic loss between 27 and 41 d after AI (18.3 vs. 16.8%) compared with control cows. The CIDR eliminated cows in estrus before the last PGF2alpha injection and decreased (P < 0.001) the proportion of cows bearing a corpus luteum (CL) at the last PGF2alpha injection because of less ovulation in response to the GnRH and greater spontaneous CL regression. Cyclic cows had greater (P = 0.03) pregnancy rates than anovulatory cows at 41 d after AI (33.8 vs. 20.4%) because of decreased (P = 0.06) late embryonic loss (16.0 vs. 30.3%). The ovulatory follicle was larger (P < 0.001) in cows in estrus, and a greater proportion of cows with follicles > or = 15 mm displayed estrus (P < 0.001) and ovulated (P = 0.05) compared with cows with follicles <15 mm. Pregnancy rates were greater (P < 0.001) for cows displaying estrus, which were related to the greater (P < 0.001) ovulation rate and decreased (P = 0.08) late embryonic loss for cows in estrus at AI. Cows that were cyclic and responded to the presynchronization protocol (high progesterone at GnRH and CL at PGF2alpha) had the highest pregnancy rates. Incorporation of a CIDR insert into a presynchronized timed AI protocol using ECP to induce estrus and ovulation did not improve pregnancy rates in lactating dairy cows. Improvements in pregnancy rates in cows treated with ECP to induce ovulation in a timed AI protocol are expected when more cows display estrus, thereby increasing ovulation rate. PMID- 15537772 TI - Effect of feed intake on endogenous losses and amino acid and energy digestibility by growing pigs. AB - An experiment was conducted to determine the effect of feed intake (FI) on endogenous losses and the digestibility of CP and AA by growing pigs. Six growing barrows (initial BW = 70.3 kg) had a T-cannula installed in the distal ileum and were used in a 6 x 6 Latin square design. A soybean meal cornstarch-based diet and a N-free diet were formulated. Chromic oxide (0.25%; as-fed basis) was included in both diets as an inert marker. Each diet was provided at three different levels of FI. Feed intake level 1 was equal to the estimated energy requirement for maintenance of the pigs, whereas levels 2 and 3 were two or three times this amount, respectively. Each experimental period lasted 7 d. The initial 4 d of each period comprised an adaptation period to the experimental diets. On d 5, fecal samples were collected, while ileal digesta were collected during two 10 h periods on d 6 and 7. Between each experimental period, a corn-soybean meal based diet (16% CP) was fed to all pigs for 7 d. The basal ileal endogenous losses (IAA(end)) of CP and AA were measured for each level of FI from pigs fed the N-free diet. Likewise, the apparent (AID) and standardized (SID) ileal digestibility coefficients for CP and AA in soybean meal were calculated for each level of FI. The total-tract digestibility coefficient of energy in the soybean meal-based diet also was calculated. The AID for CP and all indispensable AA except Lys, Met, and Thr increased (P < or =0.055) as FI increased. The IAA(end) of CP and all AA except Pro decreased linearly (P < 0.05) as FI increased when expressed as g/kg of DMI; however, the total daily IAA(end) increased as FI increased (linear, P < or =0.056) for all AA, except for Phe, Thr, Trp, Val, Cys, Gly, and Ser. The AA composition (% of CP) of endogenous protein was not affected by the level of FI, except for Arg, Thr, Pro, and Ser. As FI increased, the SID decreased linearly (P < 0.04) for CP and all AA, except Arg, Trp, Asp, Pro, and Tyr. The total-tract digestibility of energy was not influenced by the FI level. These results demonstrate that the FI level significantly influenced AID, SID, and IAA(end) for CP and AA. Therefore, pigs used to measure AA digestibility coefficients and IAA(end) should be fed at a level that is close to what is used under commercial conditions. PMID- 15537773 TI - Effects of wet corn gluten feed and intake level on diet digestibility and ruminal passage rate in steers. AB - Twelve ruminally cannulated Jersey steers (BW = 534 kg) were used in an incomplete Latin square design experiment with a 2 x 2 factorial arrangement of treatments to determine the effects of wet corn gluten feed (WCGF) and total DMI level on diet digestibility and ruminal passage rate. Treatments consisted of diets formulated to contain (DM basis) steam-flaked corn, 20% coarsely ground alfalfa hay, and either 0 or 40% WCGF offered once daily for ad libitum consumption or limited to 1.6% of BW (DM basis). Two consecutive 24-d periods were used, each consisting of 18 d for adaptation, 4 d for collection, and a 2-d in situ period. Rumens of all steers were evacuated once daily at 0, 4, 8, and 12 h after feeding. Chromic oxide (10 g/[steer*d]) was fed as a digestibility marker, and steers were pulse-dosed with Yb-labeled alfalfa hay to measure ruminal particulate passage rate. Dacron bags containing 5 g of steam-flaked corn, WCGF, or ground (2-mm screen) alfalfa hay were placed into the rumens of all steers and removed after 3, 6, 12, or 48 h. Wet corn gluten feed increased percent apparent total-tract digestion of OM (P < 0.01), NDF (P < 0.01), and starch (P < 0.03), decreased (P < 0.01) ruminal total VFA concentration, increased (P < 0.01) ruminal NH3 concentration, and increased (P < 0.01) ruminal pH. Wet corn gluten feed also increased (P < 0.01) ruminal passage rate of Yb. Limit feeding decreased (P < 0.01) percent apparent total-tract digestion of both OM and NDF, ruminal total VFA concentration (P < 0.01), and ruminal fill (P < 0.01), but increased (P < 0.01) ruminal NH3 concentration. Apparent total-tract digestion of starch was not affected (P = 0.70) by level of DMI. A DMI level x hour interaction (P < 0.01) occurred for ruminal pH. Limit feeding increased ruminal pH before and 12 h after feeding, but decreased ruminal pH 4 h after feeding compared with diets offered ad libitum. A diet x DMI level interaction (P < 0.02) occurred for in situ degradation of alfalfa hay, with dietary addition of WCGF increasing (P < 0.02) the extent of in situ alfalfa hay degradation in steers fed for ad libitum consumption. This study suggests that WCGF increases OM and NDF digestion, and that limit feeding diets once daily might depress OM and NDF digestion, possibly due to decreased stability of the ruminal environment. PMID- 15537774 TI - Effects of ammonia load on methionine utilization by growing steers. AB - Seven ruminally cannulated Holstein steers (194 +/- 16 kg) housed in metabolism crates were used in a 6 x 6 Latin square, with one additional steer, to study effects of ruminal ammonia load on methionine (Met) use. All steers received a diet based on soybean hulls (2.6 kg DM/d), ruminal infusions of 200 g/d of acetate, 200 g/d of propionate, and 50 g/d of butyrate, as well as abomasal infusion of 300 g/d of glucose to provide energy without increasing microbial protein supply, and abomasal infusions of a mixture (248 g/d) of all essential AA except Met. Treatments were arranged as a 3 x 2 factorial and included urea (0, 40, or 80 g/d) infused ruminally to supply metabolic ammonia loads and Met (2 or 5 g/d) infused abomasally. Supplementation with the greater amount of Met decreased (P < 0.05) urinary N excretion from 68.8 to 64.8 g/d and increased (P < 0.05) retained N from 22.0 to 27.5 g/d. Urea infusions linearly increased (P < 0.05) urinary N excretions, plasma urea concentrations, and urinary urea excretions, but retained N was not affected. The efficiency of deposition of supplemental Met, calculated by assuming that Met deposition is 2.0% of protein deposition (6.25 x retained N), ranged between 18 and 27% when steers received 0 or 80 g/d of urea, respectively. There were no (P > or = 0.40) effects of treatments on serum insulin or IGF-I concentrations. In our model, increasing ammonia load did not affect whole-body protein deposition in growing steers when Met was limiting. PMID- 15537775 TI - Effects of dry, wet, and rehydrated corn bran and corn processing method in beef finishing diets. AB - Two finishing trials were conducted to determine the effects of adding different types of corn bran, a component of corn gluten feed, on cattle performance. In Trial 1, 60 English crossbred yearling steers (283 +/- 6.7 kg) were used in a completely randomized design with four dietary treatments. Treatments were diets with no corn bran, dry corn bran (86% DM), wet corn bran (37% DM), and rehydrated dry bran (37% DM). Bran was fed at 40% of dietary DM. All finishing diets had (DM basis) 9% corn steep liquor with distillers solubles, 7.5% alfalfa hay, 3% tallow, and 5% supplement. Gain efficiency and ADG were greater (P < 0.01) for cattle fed no corn bran compared with all treatments containing corn bran; however, no differences were detected across corn bran types. In Trial 2, 340 English crossbred yearling steers (354 +/- 0.6 kg) were used in a randomized block design with treatments assigned based on a 2 x 4 + 2 factorial arrangement (four pens per treatment). One factor was the corn processing method used (dry rolled corn, DRC; or steam-flaked corn, SFC). The other factor was corn bran type: dry (90% DM), wet (40% DM), or dry bran rehydrated to 40 or 60% DM. Bran was fed at 30% of dietary DM, replacing either DRC or SFC. Two control diets (DRC and SFC) were fed with no added bran. All finishing diets contained (DM basis) 10% corn steep liquor with distiller's solubles, 3.5% alfalfa hay, 3.5% sorghum silage, and 5% supplement. Corn bran type did not affect DMI (P = 0.61), ADG (P = 0.53), or G:F (P = 0.10). Dry matter intake was greater (P < 0.01) by steers fed bran compared with those fed no bran, and was greater by steers fed DRC than by steers fed SFC (P < 0.01). Interactions occurred (P < 0.01) between grain source and bran inclusion for ADG and G:F. The ADG by steers fed the SFC diet without bran was greater (P < 0.01) than by steers fed SFC diets with bran, whereas the ADG by steers fed DRC diets with or without bran was similar. Daily gain was 15.2% greater (P < 0.01) by steers fed SFC without bran than by steers fed DRC without bran. Gain efficiency was 16.9% greater (P < 0.01) for steers fed SFC without bran compared with steers fed DRC without bran. In DRC and SFC diets, feeding bran decreased (P < 0.01) G:F by 5.2 and 13.8%, respectively. The moisture content of corn bran had no effect on finishing steer performance, and drying corn bran did not affect its energy value in finishing cattle diets. PMID- 15537776 TI - Effect of increasing ruminal butyrate absorption on splanchnic metabolism of volatile fatty acids absorbed from the washed reticulorumen of steers. AB - Four steers fitted with a ruminal cannula and chronic indwelling catheters in the mesenteric artery, mesenteric vein, hepatic portal vein, hepatic vein, and the right ruminal vein were used to study the absorption and metabolism of VFA from bicarbonate buffers incubated in the temporarily emptied and washed reticulorumen. Portal and hepatic vein blood flows were determined by infusion of p-aminohippurate into the mesenteric vein, and portal VFA fluxes were calibrated by infusion of isovalerate into the ruminal vein. The steers were subjected to four experimental treatments in a Latin square design with four periods within 1 d. The treatments were Control (bicarbonate buffer) and VFA buffers containing 4, 12, or 36 mmol butyrate/kg of buffer, respectively. The acetate content of the buffers was decreased with increasing butyrate to balance the acidity. The butyrate absorption from the rumen was 39, 111, and 300 +/- 4 mmol/h for the three VFA buffers, respectively. The ruminal absorption rates of propionate (260 +/- 12 mmol/h), isobutyrate (11.4 +/- 0.7 mmol/h), and valerate (17.3 +/- 0.7 mmol/h) were not affected by VFA buffers. The portal recovery of butyrate and valerate absorbed from the rumen increased (P < 0.01) with increasing butyrate absorption and reached 52 to 54 +/- 4% with the greatest butyrate absorption. The liver responded to the increased butyrate absorption with a decreasing fractional extraction of propionate and butyrate, and with the greatest butyrate absorption, the splanchnic flux was 22 +/- 1% and 18 +/- 1% of the absorbed propionate and butyrate, respectively. The increased propionate and butyrate release to peripheral tissues was followed by increased (P < 0.05) arterial concentrations of propionate (0.08 +/- 0.01 mmol/kg) and butyrate (0.07 +/- 0.01 mmol/kg). Arterial insulin concentration increased (P = 0.01) with incubation of VFA buffers compared with Control and was numerically greatest with the greatest level of butyrate absorption. We conclude that the capacity to metabolize butyrate by the ruminal epithelium and liver is limited. If butyrate absorption exceeds the metabolic capacity, it affects rumen epithelial and hepatic nutrient metabolism and affects the nutrient supply of peripheral tissues. PMID- 15537777 TI - Effect of fiber-based creep feed on intake, digestion, ruminal fermentation, and microbial efficiency in nursing calves. AB - Six Angus crossbred cow-calf pairs (653 +/- 35 kg and 157 +/- 10 kg initial BW for cows and calves, respectively) were used to evaluate the influence of a fiber based creep feed on intake, ruminal fermentation, digestion characteristics, and microbial efficiency in nursing beef calves. Cow-calf pairs were stratified by calf age and assigned randomly to one of two treatments: control (no supplement) or supplemented. Supplemented calves received 0.9 kg of a 49% soy hulls, 44% wheat middlings, 6% molasses, and 1% limestone supplement (DM basis) daily. All calves were cannulated in the rumen and duodenum and given ad libitum access to chopped brome hay (Bromus inermus L; 7.43% CP, 40.96% ADF, and 63.99% NDF; DM basis). Supplementation was initiated on May 1 (88 +/- 10.3 d calf age). Three sampling periods were conducted throughout the study (June 14 to 25, July 5 to 16, and August 9 to 20). Supplement and forage were offered at 0800 daily. Total, hay, and milk OM intakes of nursing calves were not affected by supplementation (2,014 vs. 2,328 +/- 288.8, 1,486 vs. 1,029 +/- 3,06.9, and 528 vs. 575 +/- 87.0 g/d, respectively). Milk OM intake was less (P < 0.09) in August than in June and July (635, 691, and 345 +/- 110.6 g/d for June, July, and August, respectively). A supplementation x month interaction occurred (P < 0.10) for total-tract OM digestion. Supplementation did not affect (P > 0.40) total-tract OM digestibility during June and August; however, during July, total-tract OM digestibility was lower (P = 0.03) for the control calves. Ruminal ammonia concentration, total VFA, and butyrate molar proportion increased (P < 0.05), whereas acetate proportion decreased (P = 0.01) in supplemented calves. Microbial efficiency was not influenced by supplementation (11.8 vs. 12.0 g/kg of OM truly fermented for control and supplemented calves, respectively). These data indicate that fiber based supplements can be used as creep feed without negative effects on OM intake, total-tract OM digestibility, and ruminal fermentation characteristics in nursing beef calves. PMID- 15537778 TI - Effects of supplemental protein type on intake, nitrogen balance, and site, and extent of digestion in whiteface wethers consuming low-quality grass hay. AB - Two experiments were conducted to determine the effects of supplementing ruminally degradable intake protein (DIP) or ruminally undegradable intake protein (UIP) on N balance (Exp. 1; n = 6 wethers; initial BW = 48.7 +/- 4.6 kg) and site and extent of digestion (Exp. 2; n = 5 wethers; initial BW = 36.9 +/- 3.1 kg) in whiteface wethers consuming (as-fed basis) 69% blue grama and 31% love grass hay (mixture = 7.5% CP, 73.0% NDF, 36.0% ADF [DM basis]). Treatments were 1) no supplement (Control), 2) a supplement (219 g/d, as-fed basis) low in UIP (70 g/d of CP; 24.8 g/d of UIP), and 3) a supplement (219 g/d, as-fed basis) high in UIP (70 g/d of CP; 37.1 g/d of UIP). Both experiments were replicated 3 x 3 Latin square designs, with identical feeding and supplementation. Wethers had ad libitum access to the forage mixture and fresh water, and received supplement once daily. In Exp.1, forage intake (percentage of BW) was greatest (P = 0.04) for control, but total DMI (g/d) was greatest (P = 0.05) for lambs consuming supplement. Apparent total-tract OM digestibility was numerically greater (P = 0.11) for supplemented wethers than for controls, whereas total-tract ADF digestibility tended (P = 0.08) to be greater for control wethers. Lambs fed supplements consumed and retained more (P < or = 0.01) N (% of N intake) compared with controls, but no difference (P = 0.22) was observed between low and high UIP treatments. Similar to Exp. 1, forage intake (percentage of BW) tended (P = 0.06) to be greater for control than for supplemented wethers in Exp. 2. Ruminal NDF digestibility was 16.3% greater (P = 0.02) for supplemented wethers than for controls. Postruminal NDF and N digestibilities were greatest (P < or = 0.03) for controls, but apparent OM digestibility did not differ among treatments at all sites. Duodenal N flow was greatest (P = 0.05) for high UIP and least for control wethers. Nonmicrobial N flow was greater (P = 0.02) for high UIP compared with low UIP or controls. Control wethers had greater (P = 0.05) microbial efficiency. Ruminal ammonia concentration tended (P = 0.08) to be greatest for wethers fed low UIP and least for controls, with high-UIP wethers having intermediate ammonia concentrations. Results from these experiments suggest that in lambs fed low quality forage there was no difference in apparent total-tract digestion or N balance (percentage of N intake) between lambs fed supplements that had the same CP but differed in the proportion of UIP and DIP; however, supplementing protein (regardless of UIP:DIP ratio) to wethers consuming low-quality forage increased N balance. PMID- 15537779 TI - Influence of supplemental cracked high-linoleate or high-oleate safflower seeds on site and extent of digestion in beef cattle. AB - Our objectives were to evaluate ruminal fermentation patterns, apparent ruminal biohydrogenation, and site and extent of nutrient disappearance in cattle fed supplemental cracked safflower seeds differing in 18 C fatty acid profile. Nine Angus x Gelbvieh heifers (641 +/- 9.6 kg) fitted with ruminal and duodenal cannulas were used in a triplicated 3 x 3 Latin square. Cattle were fed (OM basis) 9.1 kg of bromegrass hay and either 1) 1.8 kg of corn and 0.20 kg of soybean meal (Control); 2) 0.13 kg of soybean meal and 1.5 kg of cracked high linoleate (67.2% 18:2) safflower seeds (Linoleate); or 3) 1.5 kg of cracked high oleate (72.7% 18:1) safflower seeds (Oleate). Safflower seed supplements were formulated to provide similar quantities of N and TDN and 5% dietary fat. Single degree of freedom orthogonal contrasts (Control vs. Linoleate and Oleate; Linoleate vs. Oleate) were used to evaluate treatment effects. True ruminal OM and ruminal NDF disappearances (percentage of intake) were greater (P < or =0.02) for Control than Linoleate and Oleate. True ruminal N degradability (% of intake) was not different (P = 0.38) among treatments. Apparent ruminal biohydrogenation of dietary 18:2 was greatest (Linoleate vs. Oleate, P < 0.001) for Linoleate, whereas biohydrogenation of dietary 18:1 was greatest (Linoleate vs. Oleate, P = 0.02) for Oleate. Duodenal flow of 18:0 was least (P < 0.001) for Control but did not differ (P = 0.92) between Oleate and Linoleate. Total flow of unsaturated fatty acid to the duodenum was greatest (P < 0.001) in cattle fed safflower seeds, and was greater with Linoleate (P < 0.001) than with Oleate. Duodenal flow of 18:1 and 18:2 increased (P < 0.001) in Oleate and Linoleate, respectively. Duodenal flow of 18:1trans-11 was greater (P < 0.001) in cattle fed safflower seeds and in Linoleate than in Oleate. Postruminal disappearance of saturated fatty acids was greatest (P < 0.001) for Control; however, postruminal disappearance of total unsaturated fatty acids was greater (P = 0.002) for Linoleate vs. Oleate. Supplemental high-linoleate or high-oleate safflower seeds to cattle fed forage-based diets may negatively affect ruminal OM and fiber disappearance but not N disappearance. Provision of supplemental fat in the form of safflower seeds that are high in linoleic acid increased intestinal supply and postruminal disappearance of unsaturated fatty acids, indicating that the fatty acids apparently available for metabolism are affected by dietary fat source. PMID- 15537780 TI - Effect of field pea-based creep feed on intake, digestibility, ruminal fermentation, and performance by nursing calves grazing native range in western North Dakota. AB - Two experiments evaluated digestive and performance effects of field pea-based creep feed in nursing calf diets. In Exp.1, eight nursing steer calves (145 +/- 27 kg initial BW) with ruminal cannulas were used to evaluate effects of supplementation and advancing season on dietary composition, intake, digestion, and ruminal fermentation characteristics. Treatments were unsupplemented control (CON) and field pea-based creep (SUP; 19.1% CP, DM basis) fed at 0.45% BW (DM basis) daily. Calves grazed native range with their dams from early July through early November. Periods were 24 d long and occurred in July (JUL), August (AUG), September (SEP), and October (OCT). Experiment 2 used 80 crossbred nursing calves, 48 calves in yr 1 and 32 calves in yr 2 (yr 1 = 144 +/- 24 kg; yr 2 = 121 +/- 20 kg initial BW), to evaluate effects of field pea-based creep on calf performance. Treatments included unsupplemented control (CON); field pea-based creep feeds containing either 8% (LS); or 16% (HS) salt; and soybean meal/field pea-based creep containing (as-fed basis) 16% salt (HIPRO). Masticate samples from SUP calves in Exp.1 had greater CP (P = 0.05) than those from CON calves. Forage CP and ADIN decreased linearly with advancing season (P = 0.01 and 0.03, respectively). In vitro OM digestibility of diet masticate decreased from JUL to OCT (P < 0.01; 58.5 to 41.3%). Forage intake did not differ (P = 0.33) between treatments but increased linearly with advancing season (1.67, 1.90, 3.12, 3.38 kg/d for JUL, AUG, SEP, and OCT, respectively; P < 0.01). Milk intake (percentage of BW) did not differ (P = 0.56) between CON and SUP calves but decreased linearly (P < 0.01) with advancing season. Supplemented calves had greater (P = 0.03) total intake (g/kg of BW; forage + milk + creep) compared with CON calves. Treatment did not affect (P < 0.30) rate of in situ disappearance of forage or creep. Forage DM, CP, and creep DM disappearance rate decreased linearly (P < or = 0.02) with advancing season. Supplementation decreased (P = 0.05) ruminal pH, whereas ruminal ammonia and VFA concentrations were greater (P < or = 0.02) in SUP calves. In Exp. 2, creep-fed calves had greater ADG and final BW than CON calves (P < 0.01). Calves offered HS tended (P = 0.07) to have increased gain efficiency above CON than LS calves. Field peas can be used as an ingredient in creep feed to increase calf weight gain without negatively affecting ruminal fermentation and digestion. PMID- 15537781 TI - Effects of supplemental fat on growth performance and quality of beef from steers fed barley-potato product finishing diets: I. Feedlot performance, carcass traits, appearance, water binding, retail storage, and palatability attributes. AB - To measure the effects of dietary fat on feedlot performance, carcass characteristics, and beef appearance, moisture binding, shelf life, and palatability, 168 crossbred beef steers (317 +/- 1.0 kg) were allotted randomly, within weight blocks, to a randomized complete block design with a 3 x2 + 1 factorial arrangement of dietary treatments. Main effects were level of yellow restaurant grease (RG; 0, 3, or 6%) and level of alfalfa hay (AH; 3.5 or 7%), with the added treatment of 6% tallow and 7% AH in barley-based diets containing 15% potato by-product and 7% supplement fed for 165 d (all dietary levels on a DM basis). Dietary treatment did not (P >0.10) affect DMI, LM area, beef brightness, or beef texture. Level of RG linearly increased (P <0.05) ADG from 1.48 to 1.60 kg/d, diet NE(m) from 2.4 to 2.6 Mcal/kg, diet NE(g) from 1.7 to 1.9 Mcal/kg, and internal fat from 2.1 to 2.4%. Level of RG linearly increased (P <0.05) G:F from 0.184 to 0.202, but decreased (P <0.05) beef firmness score from 3.0 to 2.8 and fat luster score from 3.1 to 2.8. Level of AH did not (P >0.10) affect any of the measurements; however, AH interacted with level of RG on fat thickness and yield grade (linear; P <0.05), as well as marbling score and percentage of carcasses grading USDA Choice (quadratic; P <0.05). Fat thickness and yield grade increased with increasing RG level in 3.5%, but not in 7%, AH diets. In steers fed 3.5% RG, marbling scores and percentage of carcasses grading Choice were greatest when fed with 3.5% AH, and least when fed 7% AH. Steers fed tallow had lower marbling scores (P = 0.01) and percentage of carcasses grading Choice (P = 0.066) than those fed RG. Retail storage attributes, including visual and instrumental color, decreased during storage (P <0.01), but were not (P >0.10) affected by diet. Trained sensory panel scores for initial tenderness increased quadratically (P = 0.07) as dietary RG increased, but diet did not (P >0.10) affect drip loss, cooking loss, or trained sensory panel scores for sustained tenderness, initial and sustained juiciness, and beef flavor. Therefore, RG increased diet energy, improved performance, and increased carcass fatness; however, dietary fat and AH did not affect most measurements of water retention, color stability, or palatability of beef. PMID- 15537782 TI - Effects of supplemental fat on growth performance and quality of beef from steers fed barley-potato product finishing diets: II. Fatty acid composition of muscle and subcutaneous fat. AB - One hundred sixty-eight crossbred steers (317.1 +/- 1.0 kg) were used to evaluate the effects of supplemental fat in finishing diets on the fatty acid composition, including the 9,11 isomer of conjugated linoleic acid, of beef. Steers were allotted within three weight blocks to a randomized complete block design with a 3 x 2 + 1 factorial arrangement of dietary treatments. Main effects were level of yellow restaurant grease (RG; 0, 3, and 6%), and level of alfalfa hay (AH; 3.5 and 7%) with an added treatment containing 6% tallow (T) and 7% AH in barley based diets containing 15% potato by-product and 7% supplement (all dietary levels are on a DM basis) fed for an average of 165 d. Fatty acids of the LM and s.c. fat from four randomly selected steers per pen were quantified using GC after methylation with sodium methoxide. Dietary treatment did not (P > 0.10) affect total fatty acid (FA) content of the LM (143 +/- 5.2 mg/g) or fat (958 +/- 7.9 mg/g). Myristic acid increased linearly (P < 0.01) with increasing RG from 3.1 to 3.7 +/- 0.1 g/100 g of FA in muscle. Stearic acid increased linearly (P < 0.05) as RG increased in the diet, from 11.4 to 12.9 +/- 0.4 g/100 g of FA in LM and from 9.9 to 12.2 +/- 0.3 g/100 g of FA in fat. Compared with T, steers fed 6% RG had more (P < 0.05) oleic acid in LM (42.7 vs. 40.3 +/- 0.5 g/100g FA) and in fat (43.0 vs. 40.9 +/- 0.5 g/100g FA). The cis-9, trans-11 conjugated linoleic acid (CLA) increased quadratically (P < 0.01) with increasing dietary RG in LM from 0.45 to 0.64 to 0.62 +/- 0.03 g/100 g of FA and increased in fat from 0.61 to 0.84 to 0.83 +/- 0.04 g/100 g of FA. Moreover, cis-9, trans-11 CLA was higher (P < 0.05) in fat from steers fed RG compared with T (0.81 vs. 0.69 +/- 0.04 g/100 g of FA), and tended to be higher (P = 0.07) in muscle (0.62 vs. 0.54 +/- 0.03 g/100 g of FA. Feeding yellow restaurant grease increased content of cis-9, trans-11 CLA in beef without an increase total FA content. PMID- 15537783 TI - Compensatory growth improves meat tenderness in gilts but not in barrows. AB - Compensatory growth is a phenomenon observed in pigs given free access to feed following a period of restricted feeding that results in increased growth rates. Compensatory growth is believed to increase protein turnover and thereby the proteolytic potential at the time of slaughter, leading to faster tenderization rates of meat. Nine litters of three gilts and three barrows were allocated within litter and gender to three dietary treatment groups. Pigs had ad libitum access to feed from d 28 to slaughter at d 140 (ALA) or were restricted to 69% ad libitum from d 28 to d 80 or 90, and then given ad libitum access to the diet until slaughter at d 140 (RA80 and RA90, respectively). Pigs in the RA80 and RA90 treatment groups had a 9.7% higher (P < or = 0.001) fractional growth rate in the second feeding period than those in the ALA group. Growth rate was correlated to the activity of m-calpain (r = 0.37; P < or = 0.01), beta-glucuronidase (r = 0.48; P < or = 0.001), and cathepsins B (r = 0.47; P < or = 0.001) and B+L (r = 0.31; P < or = 0.04). The LM of RA80-gilts received higher tenderness scores than the LM of ALA gilts, but tenderness scores were similar among barrows regardless of treatment (gender x treatment; P = 0.02). Conversely, tenderness scores were higher for the biceps femoris of ALA barrows than either ALA gilts or RA90 barrows (gender x treatment; P = 0.02). Desmin and troponin-T degradation, as well as myofibrillar fragmentation index, of the LM were not (P > or = 0.24) affected by treatment. No dietary treatment effects were observed on the activities of mu-calpain (P = 0.15), m-calpain (P = 0.74), or calpastatin (P = 0.91) at slaughter. The cathepsin inhibitors, cystatins, tended to be increased (P = 0.06) in RA80 and RA90 pigs. Sarcomere length was longer (P = 0.003) in the LM of gilts than barrows. Barrows in the RA80 group had lower i.m. fat concentrations than ALA; however, no differences were found in the LM of gilts (gender x treatment; P = 0.03). The underlying hypothesis that compensatory growth leads to an increased proteolytic potential at the time of slaughter could not be verified in this study. PMID- 15537784 TI - Effect of diet composition and feeding pattern on the prececal digestibility of starches from diverse botanical origins measured with the mobile nylon bag technique in horses. AB - This trial was conducted to determine the extent of prececal starch digestibility depending on the botanical origin of starch and on diet characteristics (i.e., composition and feeding pattern). The prececal disappearance of six substrates (oats, barley, corn, horse bean, potato, and wheat) was measured in four cannulated horses fed (as-fed basis) 11.8 g/kg BW of a high-fiber (HF) or high starch (HS) pelleted feed and 10.0 g/kg BW of meadow hay using the mobile bag technique (MBT). The daily feeding pattern was either three meals (two meals of pellets and one meal of hay) or five meals (three meals of pellets and two meals of hay). The experimental procedure was a 2 x 2 factorial arrangement tested in a Latin square design. After 2 wk of adaptation to the diet, collections were made on 5 d. Thirty nylon bags, composed of five bags of each substrate, were intubated to each horse during the ingestion of the morning meal. Bags were collected in the cecum, using a magnet, at 9 h postintubation. In spite of strong interindividual differences, approximately 80% of the intubated bags were collected. On average, the mean retention time of the bags was 6.2 h (+/-0.17). Regardless of the feeding pattern, the transit of the bags was faster when the fiber content of the diet was higher (P = 0.003). Likewise, regardless of the meal composition, transit was also faster when the ration was split into five daily meals (P = 0.001). The DM disappearance, corrected with particulate losses (DMD(c)), differed depending on the substrate tested (33.5, 57.1, 63.8, 67.7, 78.6, and 86.2% for potato, horse bean, oats, barley, corn, and wheat, respectively; P = 0.001). The DMD(c) of corn, barley, and potato was higher when HS was fed (P = 0.020); regardless of the substrate, DMD(c) was higher with five daily meals (P = 0.001). The starch disappearance (StarchD(c)) was different depending on the substrate (P = 0.001; 36.1, 71.2, 86.6, 89.2, 99.0, and 99.7% for potato, horse bean, barley, corn, wheat, and oats, respectively). Whatever the substrate, StarchD(c) was higher when HS was fed (P = 0.007), but it was not affected by the feeding pattern of the diet. Although passage rate was modified and feed intake was different, the botanical origin of starch was the main factor that affected prececal starch disappearance in horses. PMID- 15537785 TI - Comparison of wheat gluten and spray-dried animal plasma in diets for nursery pigs. AB - Five experiments were conducted to determine the effects of different wheat gluten (WG) sources (Source 1 = enzymatically hydrolyzed, Source 2 = nonmodified ring-dried, Source 3 = spray-dried, and Source 4 = flash-dried) on growth performance of nursery pigs compared with soybean meal (SBM), spray-dried animal plasma (SDAP), or other specialty protein sources. In Exp. 1, pigs (n = 220, initially 6.1 +/- 2.5 kg) were fed a control diet containing (as-fed basis) 6% SDAP or WG Source 1 or 2. The WG and l-lysine*HCl replaced 50 or 100% of the SDAP. From d 0 to 21, increasing WG (either source) decreased ADG and ADFI (linear, P < 0.01), but improved (linear, P < 0.02) G:F. In Exp. 2, pigs (n = 252, initially 6.2 +/- 3.0 kg) were fed a negative control diet containing no SDAP or WG, diets containing (as-fed basis) 9% WG Source 1 or 5% SDAP, or combinations of WG and SDAP where WG and l-lysine*HCl replaced 25, 50, or 75% of SDAP. From d 0 to 14, pigs fed increasing WG had decreased ADG (linear, P < 0.05). In Exp. 3, pigs (n = 240, initially 7.0 +/- 2.5 kg) were fed a negative control diet, a diet containing (as-fed basis) either 3, 6, 9, or 12% WG Source 3, or a positive control diet containing 5% SDAP. The diets containing 9% WG and 5% SDAP had the same amount of SBM. From d 0 to 7, pigs fed 5% SDAP had greater (P < 0.04) ADG than pigs fed the diet containing 9% WG. From d 0 to 14, increasing WG had no effect on ADG, ADFI, or G:F. In Exp. 4, pigs (n = 200, initially 6.0 +/- 2.4 kg) were fed a negative control diet, the control diet with (as-fed basis) 4.5 or 9.0% WG Source 1, or the control diet with 2.5 or 5.0% SDAP. Diets containing WG and SDAP had similar SBM levels. From d 0 to 7 and 0 to 14, increasing SDAP tended to improve (linear, P < 0.06) ADG, but increasing WG had no effect. In Exp. 5, 170 barrows and gilts (initially 7.5 +/- 2.8 kg) were used to determine the effects of WG Source 1 and 4 compared with select Menhaden fish meal or spray-dried blood cells and a negative control diet (SBM) on the growth performance of nursery pigs from d 5 to 26 postweaning (d 0 to 21 of experiment). No differences were found in ADG or G:F, but pigs fed the diet containing (as-fed basis) 2.5% spray-dried blood cells had greater ADFI than pigs fed the negative control from d 0 to 21. Wheat gluten source had no effect on ADG, ADFI, or G:F. The results of these studies suggest that increasing WG in diets fed immediately after weaning did not improve growth performance relative to SBM or SDAP. PMID- 15537786 TI - Effects of feeding strategy and age on live animal performance, carcass characteristics, and economics of short-term feeding programs for culled beef cows. AB - To evaluate production and economic effects of feeding management strategy and age on intensively managed culled beef cows, a study was conducted using 125 cows of British breeding blocked by age (Young = 3 and 4 yr olds; LowMid = 5 and 6 yr olds; HighMid = 7 and 8 yr olds; and Aged = 9 yr and older) and assigned to one of three steam-flaked corn based feeding strategies. Treatments were as follows: Conservative (CSV), 30% roughage throughout; Standard (STD), decrease roughage from 30 to 10% over 20 d; and Aggressive (AGR), decrease roughage from 30 to 10% over 10 d. There were four pens per treatment in a randomized complete block design. Cows were fed for a total of 54 d, and BW was measured on d 0, 14, 28, 42, and 54. Half the cows from each pen were randomly selected and slaughtered at a commercial abattoir, and carcass data were collected. Average daily gain, daily DMI, and G:F during each weigh period and across the entire feeding period were calculated. Over the 54-d feeding period, strategies that employed more energy dense diets numerically increased ADG (1.28, 1.63, and 1.55 +/- 0.14 kg/d for CSV, STD, and AGR; P = 0.26) and decreased DMI (11.91, 10.74, and 10.89 +/- 0.27 kg/d for CSV, STD, and AGR; P = 0.05), such that G:F was lower for CSV than for STD or AGR (0.105, 0.150, and 0.141 +/- 0.010; P = 0.05). Carcass weight was least for the CSV strategy (298 kg) and greatest for STD (328 kg); AGR resulted in intermediate carcass weight (317 +/- 6 kg; P = 0.04). Total cost of gain was over 30% greater for CSV strategy than for STD or AGR strategies (P < 0.01). In many cases, block effects (age) had a greater effect on responses than treatments. Average daily gain, DMI, and G:F decreased linearly with age (P < 0.01). Hot carcass weight, dressing percent, and fat thickness decreased linearly with age (P < 0.03); yield grade decreased and carcass maturity attributes increased linearly with age (P < 0.02). Performance and intake differences resulted in linear increases in total cost of gain (P < 0.01) and breakeven price (P = 0.03) with increasing age. These data indicate advantages to more aggressive feeding management strategies for culled beef cows, although maximal intake may be achieved with higher-roughage diets. Despite management effects, an increase in market price above purchase price may be required for intensive feeding of culled beef cows to be a profitable enterprise. PMID- 15537787 TI - High linoleic acid safflower seed supplementation for gestating ewes: effects on ewe performance, lamb survival, and brown fat stores. AB - Objectives of this study were to determine whether feeding high-linoleic safflower seed to gestating ewes increases cold tolerance and survival in lambs, and whether brown adipose tissue (BAT) stores in lambs are affected by prepartum safflower seed supplementation. In Trial 1, 234 gestating ewes (122 in yr 1 and 112 in yr 2; 75.5 and 81.2 +/- 0.6 kg initial BW for yr 1 and 2, respectively) were allotted randomly to one of two dietary treatments (four pens*treatment( 1)*yr(-1)). Ewes were fed alfalfa-based diets containing (DM basis) either 2.8 (LF) or 5.7% (HF) dietary fat beginning 55 (yr 1) and 42 (yr 2) +/- 1 d prepartum. In Trial 2, 40 Rambouillet cross ewes gestating twins (82.9 +/- 1.7 kg BW) were used in 2 yr (20/yr) and were fed diets containing (DM basis) either 1.9 (LF) or 4.9% (HF) dietary fat beginning 53.4 +/- 1.4 d prepartum. The basal diet was 37.5% each of grass and alfalfa hays and 25% corn silage (DM basis). Cracked safflower seeds (18% CP, 32% fat, 25.6% linoleic acid; DM basis) were used as the supplement in HF, whereas safflower meal and corn were used as the supplement in LF for both trials. At parturition, one lamb from each ewe was selected randomly for slaughter. Perirenal (PR) and pericardial (PC) BAT was excised and weighed, and the carcass was frozen for compositional analysis. In Trial 1, more lambs from HF 0.03; 15.4 vs. 5.8 +/- 2.8%), and dams survived (P = 0.03; 88.4 vs. 78.3 +/- 2.9%), fewer died due to starvation (P = there was a tendency for fewer to die due to pneumonia (P = 0.07; 0.0 vs. 1.7 +/- 0.6%). Ewes fed HF tended to wean more lambs per ewe (P = 0.09; 1.4 vs. 1.2 +/- 0.06) but had similar lamb weight weaned per ewe (P = 0.51; 23.1 +/- 1.22 kg). In Trial 2, prepartum ewe plasma NEFA and glucose concentrations increased with advancing gestation (P < 0.001). Lamb rectal temperature tended (P = 0.08) to be higher in LF lambs and tended (P = 0.06) to increase following parturition. Perirenal BAT weight did not differ among treatments (33.01 +/- 1.66 g; P = 0.28; 0.62 +/- 0.30% BW; P = 0.60). Lambs from LF dams tended (P = 0.08) to have greater PC BAT weight; however, the effect was not significant when expressed as a percentage of BW (0.13 +/- 0.007; P = 0.98). High-linoleic safflower seeds fed during the last 45 d of gestation may be beneficial in improving lamb survivability. Our data do not indicate this response was a result of increased BAT stores. More research is necessary to determine mechanisms that enhance lamb survival when high-linoleic saf-flower seed is fed during gestation. PMID- 15537788 TI - Effects of increasing level of supplemental barley on forage intake, digestibility, and ruminal fermentation in steers fed medium-quality grass hay. AB - Objectives of this research were to evaluate effects of increasing level of barley supplementation on forage intake, digestibility, and ruminal fermentation in beef steers fed medium-quality forage. Four crossbred ruminally cannulated steers (average initial BW = 200 +/- 10 kg) were used in a 4 x 4 Latin square design. Chopped (5 cm) grass hay (10% CP) was offered ad libitum with one of four supplements. Supplements included 0, 0.8, 1.6, or 2.4 kg of barley (DM basis) and were fed in two equal portions at 0700 and 1600. Supplements were fed at levels to provide for equal intake of supplemental protein with the addition of soybean meal. Forage intake (kg and g/kg BW) decreased linearly (P < 0.01), and total intake increased linearly (P < 0.03) with increasing level of barley supplementation. Digestible OM intake (g/kg BW) increased linearly (P < 0.01) with increasing level of barley supplementation; however, the majority of this response was observed with 0.8 kg of barley supplementation. Treatments had only minor effects on ruminal pH, with decreases occurring at 15 h after feeding in steers receiving 2.4 kg of barley supplementation. Total-tract digestibility of DM, OM, NDF, and CP were increased (P < 0.04) with barley supplementation; however, ADF digestibility was decreased by 1.6 and 2.4 kg of barley supplementation compared with controls. Ruminal ammonia concentrations decreased linearly (P < 0.01) at 1 through 15 h after feeding. Total ruminal VFA concentrations were not altered by dietary treatments. Ruminal proportions of acetate and butyrate decreased (P < 0.10) in response to supplementation. Rate, lag, and extent (72 h) of in situ forage degradability were unaffected by treatment. Generally, these data are interpreted to indicate that increasing levels of barley supplementation decrease forage intake, increase DM, OM, and NDF digestibility, and indicate alteration of the ruminal environment and fermentation patterns. PMID- 15537789 TI - Structural analysis of the lipopolysaccharide from Pasteurella multocida genome strain Pm70 and identification of the putative lipopolysaccharide glycosyltransferases. AB - Pasteurella multocida is an important multispecies veterinary pathogen. The cell surface lipopolysaccharide (LPS) is an important virulence factor and forms the basis of the serotyping scheme, although little structural information about it is known. The structure of the LPS from the Pasteurella multocida genome strain Pm70 was elucidated in this study. The LPS was subjected to a variety of degradative procedures. The structures of the purified products were established by monosaccharide and methylation analyses, NMR spectroscopy, and mass spectrometry. The structure of the core oligosaccharide was determined on the basis of the combined data from these experiments. Identification of the core oligosaccharide structure enabled a search for glycosyltransferase homologs in the Pm70 genome and revealed a clustering of the genes putatively responsible for outer core oligosaccharide biosynthesis. PMID- 15537790 TI - Human EDEM2, a novel homolog of family 47 glycosidases, is involved in ER associated degradation of glycoproteins. AB - In the endoplasmic reticulum (ER), misfolded proteins are retrotranslocated to the cytosol and degraded by the proteasome in a process known as ER-associated degradation (ERAD). Early in this pathway, a proposed lumenal ER lectin, EDEM, recognizes misfolded glycoproteins in the ER, disengages the nascent molecules from the folding pathway, and facilitates their targeting for disposal. In humans there are a total of three EDEM homologs. The amino acid sequences of these proteins are different from other lectins but are closely related to the Class I mannosidases (family 47 glycosidases). In this study, we characterize one of the EDEM homologs from Homo sapiens, which we have termed EDEM2 (C20orf31). Using recombinantly generated EDEM2, no alpha-1,2 mannosidase activity was observed. In HEK293 cells, recombinant EDEM2 is localized to the ER where it can associate with misfolded alpha1-antitrypsin. Overexpression of EDEM2 accelerates the degradation of misfolded alpha1-antitrypsin, indicating that the protein is involved in ERAD. PMID- 15537791 TI - Synthesis, preliminary characterization, and application of novel surfactants from highly branched xyloglucan oligosaccharides. AB - A novel class of nonionic, carbohydrate-based surfactants has been synthesized from the plant polysaccharide xyloglucan. Enzymatic hydrolysis of xyloglucan yielded a series of well-defined, highly branched oligosaccharides that, following reductive amination, were readily conjugated with fatty acids bearing C8 to C18 chains under mild conditions. The critical micelle concentration, determined by tensiometry and dye-inclusion measurements, showed a typical dependence on acyl chain length and was sensitive to the degree of galactosylation of the head group. Several compounds from this new group of surfactants, especially those with C14 and C16 chains, were useful for the extraction of membrane-bound enzyme markers from different plant cell compartments in catalytically active form. PMID- 15537792 TI - Isolation, characterization, and extra-embryonic secretion of the Xenopus laevis embryonic epidermal lectin, XEEL. AB - The Xenopus laevis embryonic epidermal lectin (XEEL) is a novel member of a group of lectins including mammalian intelectins, frog oocyte cortical granule lectins, and plasma lectins in lower vertebrates and ascidians. We isolated the XEEL protein from the extract of tailbud embryos by affinity chromatography on a galactose-Sepharose column. The XEEL protein is a homohexamer of 43-kDa N glycosylated peptide subunits linked by disulfide bonds. It requires Ca(2+) for saccharide binding and shows a higher affinity to pentoses than hexoses and disaccharides. HEK-293T cells transfected with an expression vector containing the XEEL cDNA secrete into the culture medium the recombinant XEEL (rXEEL) that is similar to the purified XEEL in its molecular nature and saccharide-binding properties. Substitution of Asn-192 to Gln removed the N-linked carbohydrate and inhibited secretion of rXEEL but did not abolish the activity to bind to galactose-Sepharose. The embryo's XEEL content, as estimated by western blot analyses, increases during neurula/tailbud stages and declines after 1 week postfertilization. Immunofluorescence and immuno-electron microscopic analyses showed localization of the XEEL protein in a typical secretory granule pathway of nonciliated epidermal cells. When tailbud embryos were cultured in the standard medium, XEEL was accumulated in the medium, indicating secretion of XEEL into the environmental water. The rate of XEEL secretion greatly increased at around the hatching stage and stayed at a high level during the first week after hatching. XEEL may have a role in innate immunity to protect embryos and larvae against pathogenic microorganisms in the environmental water. PMID- 15537793 TI - DO outlines steps to malpractice reform. PMID- 15537794 TI - The elephant in the room: does OMT have proved benefit? PMID- 15537795 TI - More about the use of OMT during influenza epidemics. PMID- 15537797 TI - Clinical trial registration, a needed addition to the research process. PMID- 15537798 TI - Prevalence of frontal plane pelvic postural asymmetry--part 1. AB - Despite 80 years of study, questions of how leg length difference relates to recurrent pain and somatic dysfunction remain controversial. The authors hypothesize that a correlation exists between leg length inequality and back pain. They further hypothesize that if common compensatory patterns described in classic osteopathic medical literature exist, these patterns should interact with the pelvic postural asymmetry patterns of Lloyd and Eimerbrink in a predictable, most probable, and congruent fashion. This article reviews the osteopathic medical, as well as the allopathic medical and chiropractic literature for studies that meet criteria for evidence-based comparison. Using lumbar radiographic studies produced with subjects standing, the authors examined the prevalence of six types of pelvic postural asymmetry in a consecutive case series of 421 patients with low back pain. Establishing the frequency of pelvic postural asymmetry patterns is a necessary first step in creating an evidence-based foundation to further clarify postural compensatory patterns. Various correlations between and within these patterns are identified. PMID- 15537799 TI - Outpatient Osteopathic Single Organ System Musculoskeletal Exam Form series: validation of the Outpatient Osteopathic SOS Musculoskeletal Exam Form, a new standardized medical record. AB - The authors validate the Outpatient Osteopathic SOS (Single Organ System) Musculoskeletal Exam Form (SOS MSEF), a 1-page form contained within the 4-page Outpatient Osteopathic Single Organ System Musculoskeletal Exam Form Series (SOS FS). Handwritten physician progress notes (PPNs) in the medical record (considered to be the "gold standard" for clinical records) were compared with information placed on the SOS MSEF for the same patient encounter. Data recorded by 14 trained and certified investigators on the standardized SOS MSEF-which was designed for use with the previously validated Outpatient Osteopathic SOAP (Subjective, Objective, Assessment, Plan) Note Form (SNF)-was compared with data recorded by the same investigators in PPNs. The authors compared the accuracy and efficiency of physicians recording musculoskeletal information in these two formats for 165 patient encounters. Descriptive statistics and t tests were used to compare data recorded after patient encounters. Ninety-seven variables input from the PPNs or SOS MSEFs were significantly different at the P < or = .05 level, whereas 38 variables were not. Insufficient data was recorded for a determination of significance in 3 variables. For 121 variables, more data were recorded using the SOS MSEFs than PPNs; for 84 variables, the amount of data recorded exceeded twice that recorded using PPNs. For 10 variables, more data were recorded in PPNs; however, these differences were not significant. The authors conclude that the SOS MSEF is superior to PPNs for recording patient encounter data in the osteopathic care setting. Moreover, they argue that the use of the validated SOS MSEF nationwide would ensure that osteopathic physicians would be recording data in a similar manner for uniform insurance claim coding, easy tracking of physicians-in-training and patient outcomes, and data collection for future research. PMID- 15537801 TI - Prediction of site-specific amino acid distributions and limits of divergent evolutionary changes in protein sequences. AB - We derive an analytic expression for site-specific stationary distributions of amino acids from the structurally constrained neutral (SCN) model of protein evolution with conservation of folding stability. The stationary distributions that we obtain have a Boltzmann-like shape, and their effective temperature parameter, measuring the limit of divergent evolutionary changes at a given site, can be predicted from a site-specific topological property, the principal eigenvector of the contact matrix of the native conformation of the protein. These analytic results, obtained without free parameters, are compared with simulations of the SCN model and with the site-specific amino acid distributions obtained from the Protein Data Bank. These results also provide new insights into how the topology of a protein fold influences its designability, i.e., the number of sequences compatible with that fold. The dependence of the effective temperature on the principal eigenvector decreases for longer proteins, as a possible consequence of the fact that selection for thermodynamic stability becomes weaker in this case. PMID- 15537802 TI - Evolution of duplicate control regions in the mitochondrial genomes of metazoa: a case study with Australasian Ixodes ticks. AB - To investigate the evolution pattern and phylogenetic utility of duplicate control regions (CRs) in mitochondrial (mt) genomes, we sequenced the entire mt genomes of three Ixodes species and part of the mt genomes of another 11 species. All the species from the Australasian lineage have duplicate CRs, whereas the other species have one CR. Sequence analyses indicate that the two CRs of the Australasian Ixodes ticks have evolved in concert in each species. In addition to the Australasian Ixodes ticks, species from seven other lineages of metazoa also have mt genomes with duplicate CRs. Accumulated mtDNA sequence data from these metazoans and two recent experiments on replication of mt genomes in human cell lines with duplicate CRs allowed us to re-examine four intriguing questions about the presence of duplicate CRs in the mt genomes of metazoa: (1) Why do some mt genomes, but not others, have duplicate CRs? (2) How did mt genomes with duplicate CRs evolve? (3) How could the nucleotide sequences of duplicate CRs remain identical or very similar over evolutionary time? (4) Are duplicate CRs phylogenetic markers? It appears that mt genomes with duplicate CRs have a selective advantage in replication over mt genomes with one CR. Tandem duplication followed by deletion of genes is the most plausible mechanism for the generation of mt genomes with duplicate CRs. Once duplicate CRs occur in an mt genome, they tend to evolve in concert, probably by gene conversion. However, there are lineages where gene conversion may not always occur, and, thus, the two CRs may evolve independently in these lineages. Duplicate CRs have much potential as phylogenetic markers at low taxonomic levels, such as within genera, within families, or among families, but not at high taxonomic levels, such as among orders. PMID- 15537803 TI - Identifying signatures of selection at the enhancer of split neurogenic gene complex in Drosophila. AB - The Enhancer of split gene complex (E(spl)-C) is one of the more highly annotated gene regions in Drosophila, and the 12 genes within the complex help determine the spacing and patterning of adult bristles. Any E(spl)-C coding, transcribed, or cis-regulatory regions experiencing nonneutral evolution are strong candidates to harbor polymorphisms contributing to naturally occurring variation in bristle number. We confirm that the E(spl)-C is strongly conserved and show that 74% of regulatory elements previously identified in D. melanogaster are conserved in D. pseudoobscura. Regulatory elements in enhancer regions show lower nucleotide diversity and more rare polymorphisms compared with adjacent nonregulatory DNA, suggesting they are under purifying selection, and these effects are particularly pronounced when considering only conserved regulatory elements. The ratio of polymorphism to divergence was significantly different between binding sites and nonbinding sites for transcription factors within enhancer regions, suggesting the action of some form of selection. Too few polymorphisms in regions of the 3' UTR harboring regulatory motifs prevents adequate comparison of diversity and the polymorphism frequency spectrum between 3' UTR motif and nonmotif sequence. We identified at least two broad regions of the gene complex showing strong population subdivision among four populations, which is suggestive of local adaptation or background selection. Finally, two regions of the E(spl)-C exhibit low nucleotide diversity, a high level of rare polymorphisms, and an increase in linkage disequilibrium, which together suggest the action of positive selection. Notably, the gene m2 shows a significant deviation from neutrality by the McDonald-Kreitman test and resides in one of the two regions putatively experiencing a selective sweep. All sites in regions apparently visible to various selective forces are candidates for future work to determine their phenotypic effects. PMID- 15537804 TI - Inverse relationship between evolutionary rate and age of mammalian genes. AB - A large number of genes is shared by all living organisms, whereas many others are unique to some specific lineages, indicating their different times of origin. The availability of a growing number of eukaryotic genomes allows us to estimate which mammalian genes are novel genes and, approximately, when they arose. In this article, we classify human genes into four different age groups and estimate evolutionary rates in human and mouse orthologs. We show that older genes tend to evolve more slowly than newer ones; that is, proteins that arose earlier in evolution currently have a larger proportion of sites subjected to negative selection. Interestingly, this property is maintained when a fraction of the fastest-evolving genes is excluded or when only genes belonging to a given functional class are considered. One way to explain this relationship is by assuming that genes maintain their functional constraints along all their evolutionary history, but the nature of more recent evolutionary innovations is such that the functional constraints operating on them are increasingly weaker. Alternatively, our results would also be consistent with a scenario in which the functional constraints acting on a gene would not need to be constant through evolution. Instead, starting from weak functional constraints near the time of origin of a gene-as supported by mechanisms proposed for the origin of orphan genes-there would be a gradual increase in selective pressures with time, resulting in fewer accepted mutations in older versus more novel genes. PMID- 15537805 TI - Genome-wide analysis of core cell cycle genes in the unicellular green alga Ostreococcus tauri. AB - The cell cycle has been extensively studied in various organisms, and the recent access to an overwhelming amount of genomic data has given birth to a new integrated approach called comparative genomics. Comparing the cell cycle across species shows that its regulation is evolutionarily conserved; the best-known example is the pivotal role of cyclin-dependent kinases in all the eukaryotic lineages hitherto investigated. Interestingly, the molecular network associated with the activity of the CDK-cyclin complexes is also evolutionarily conserved, thus, defining a core cell cycle set of genes together with lineage-specific adaptations. In this paper, we describe the core cell cycle genes of Ostreococcus tauri, the smallest free-living eukaryotic cell having a minimal cellular organization with a nucleus, a single chloroplast, and only one mitochondrion. This unicellular marine green alga, which has diverged at the base of the green lineage, shows the minimal yet complete set of core cell cycle genes described to date. It has only one homolog of CDKA, CDKB, CDKD, cyclin A, cyclin B, cyclin D, cyclin H, Cks, Rb, E2F, DP, DEL, Cdc25, and Wee1. We have also added the APC and SCF E3 ligases to the core cell cycle gene set. We discuss the potential of genome-wide analysis in the identification of divergent orthologs of cell cycle genes in different lineages by mining the genomes of evolutionarily important and strategic organisms. PMID- 15537806 TI - CpG mutation rates in the human genome are highly dependent on local GC content. AB - CpG dinucleotides mutate at a high rate because cytosine is vulnerable to deamination, cytosines in CpG dinucleotides are often methylated, and deamination of 5-methylcytosine (5mC) produces thymidine. Previous experiments have shown that DNA melting is the rate-limiting step in cytosine deamination. Here we show, through the analysis of human single-nucleotide polymorphisms (SNPs), that the mutation rate produced by 5mC deamination is highly dependent on local GC content. In fact, linear regression analysis showed that the log(10) of the 5mC mutation rates (inferred from SNP frequencies) had slopes of -3 when graphed with respect to the GC content of neighboring sequences. This is the ideal slope that would be expected if the correlation between CpG underrepresentation and GC content had been solely caused by DNA melting. Moreover, this same result was obtained regardless of the SNP locations (all SNPs versus only SNPs in noncoding intergenic regions, excluding CpG islands) and regardless of the lengths over which GC content was calculated (SNP sequences with a modal length of 564 bp versus genomic contigs with a modal length of 163 kb). Several alternative interpretations are discussed. PMID- 15537807 TI - "Species" radiations of symbiotic dinoflagellates in the Atlantic and Indo Pacific since the Miocene-Pliocene transition. AB - Endosymbiotic dinoflagellates, or "zooxanthellae," are required for the survival of a diverse community of invertebrates that construct and dominate shallow, tropical coral reef ecosystems. Molecular systematics applied to this once understudied symbiont partner, Symbiodinium spp., divide the group into divergent lineages or subgeneric "clades." Within each clade, numerous closely related "types," or species, exhibit distinctive host taxon, geographic, and/or environmental distributions. This diversity is greatest in clade C, which dominates the Indo-Pacific host fauna and shares dominance in the Atlantic Caribbean with clade B. Two "living" ancestors in this group, C1 and C3, are common to both the Indo-Pacific and Atlantic-Caribbean. With these exceptions, each ocean possesses a diverse clade C assemblage that appears to have independently evolved (adaptively radiated) through host specialization and allopatric differentiation. This phylogeographic evidence suggests that a worldwide selective sweep of C1/C3, or their progenitor, must have occurred before both oceans separated. The probable timing of this event corresponds with the major climactic changes and low CO(2) levels of the late Miocene and/or early Pliocene. Subsequent bursts of diversification have proceeded in each ocean since this transition. An ecoevolutionary expansion to numerous and taxonomically diverse hosts by a select host-generalist symbiont followed by the onset of rapid diversification suggests a radical process through which coral-algal symbioses respond and persist through the vicissitudes of planetary climate change. PMID- 15537808 TI - The influence of mutation, recombination, population history, and selection on patterns of genetic diversity in Neisseria meningitidis. AB - Patterns of genetic diversity within populations of human pathogens, shaped by the ecology of host-microbe interactions, contain important information about the epidemiological history of infectious disease. Exploiting this information, however, requires a systematic approach that distinguishes the genetic signal generated by epidemiological processes from the effects of other forces, such as recombination, mutation, and population history. Here, a variety of quantitative techniques were employed to investigate multilocus sequence information from isolate collections of Neisseria meningitidis, a major cause of meningitis and septicemia world wide. This allowed quantitative evaluation of alternative explanations for the observed population structure. A coalescent-based approach was employed to estimate the rate of mutation, the rate of recombination, and the size distribution of recombination fragments from samples from disease-associated and carried meningococci obtained in the Czech Republic in 1993 and a global collection of disease-associated isolates collected globally from 1937 to 1996. The parameter estimates were used to reject a model in which genetic structure arose by chance in small populations, and analysis of molecular variation showed that geographically restricted gene flow was unlikely to be the cause of the genetic structure. The genetic differentiation between disease and carriage isolate collections indicated that, whereas certain genotypes were overrepresented among the disease-isolate collections (the "hyperinvasive" lineages), disease-associated and carried meningococci exhibited remarkably little differentiation at the level of individual nucleotide polymorphisms. In combination, these results indicated the repeated action of natural selection on meningococcal populations, possibly arising from the coevolutionary dynamic of host-pathogen interactions. PMID- 15537809 TI - Codon bias signatures, organization of microorganisms in codon space, and lifestyle. AB - New and simple numerical criteria based on a codon adaptation index are applied to the complete genomic sequences of 80 Eubacteria and 16 Archaea, to infer weak and strong genome tendencies toward content bias, translational bias, and strand bias. These criteria can be applied to all microbial genomes, even those for which little biological information is known, and a codon bias signature, that is the collection of strong biases displayed by a genome, can be automatically derived. A codon bias space, where genomes are identified by their preferred codons, is proposed as a novel formal framework to interpret genomic relationships. Principal component analysis confirms that although GC content has a dominant effect on codon bias space, thermophilic and mesophilic species can be identified and separated by codon preferences. Two more examples concerning lifestyle are studied with linear discriminant analysis: suitable separating functions characterized by sets of preferred codons are provided to discriminate: translationally biased (hyper)thermophiles from mesophiles, and organisms with different respiratory characteristics, aerobic, anaerobic, facultative aerobic and facultative anaerobic. These results suggest that codon bias space might reflect the geometry of a prokaryotic "physiology space." Evolutionary perspectives are noted, numerical criteria and distances among organisms are validated on known cases, and various results and predictions are discussed both on methodological and biological grounds. PMID- 15537810 TI - Molecular clocks do not support the Cambrian explosion. AB - The fossil record has long supported the view that most animal phyla originated during a brief period approximately 520 MYA known as the Cambrian explosion. However, molecular data analyses over the past 3 decades have found deeper divergences among animals (approximately 800 to 1,200 MYA), with and without the assumption of a global molecular clock. Recently, two studies have instead reported time estimates apparently consistent with the fossil record. Here, we demonstrate that methodological problems in these studies cast doubt on the accuracy and interpretations of the results obtained. In the study by Peterson et al., young time estimates were obtained because fossil calibrations were used as maximum limits rather than as minimum limits, and not because invertebrate calibrations were used. In the study by Aris-Brosou and Yang, young time estimates were obtained because of problems with rate models and other methods specific to the study, and not because Bayesian methods were used. This also led to many anomalous findings in their study, including a primate-rodent divergence at 320 MYA. With these results aside, molecular clocks continue to support a long period of animal evolution before the Cambrian explosion of fossils. PMID- 15537811 TI - Modeling sleep and wakefulness in the thalamocortical system. AB - When the brain goes from wakefulness to sleep, cortical neurons begin to undergo slow oscillations in their membrane potential that are synchronized by thalamocortical circuits and reflected in EEG slow waves. To provide a self consistent account of the transition from wakefulness to sleep and of the generation of sleep slow waves, we have constructed a large-scale computer model that encompasses portions of two visual areas and associated thalamic and reticular thalamic nuclei. Thousands of model neurons, incorporating several intrinsic currents, are interconnected with millions of thalamocortical, corticothalamic, and both intra- and interareal corticocortical connections. In the waking mode, the model exhibits irregular spontaneous firing and selective responses to visual stimuli. In the sleep mode, neuromodulatory changes lead to slow oscillations that closely resemble those observed in vivo and in vitro. A systematic exploration of the effects of intrinsic currents and network parameters on the initiation, maintenance, and termination of slow oscillations shows the following. 1) An increase in potassium leak conductances is sufficient to trigger the transition from wakefulness to sleep. 2) The activation of persistent sodium currents is sufficient to initiate the up-state of the slow oscillation. 3) A combination of intrinsic and synaptic currents is sufficient to maintain the up-state. 4) Depolarization-activated potassium currents and synaptic depression terminate the up-state. 5) Corticocortical connections synchronize the slow oscillation. The model is the first to integrate intrinsic neuronal properties with detailed thalamocortical anatomy and reproduce neural activity patterns in both wakefulness and sleep, thereby providing a powerful tool to investigate the role of sleep in information transmission and plasticity. PMID- 15537812 TI - Dynamic properties of antennal responses to pheromone in two moth species. AB - Dynamic properties of pheromone plumes are behaviorally important in some moths for inducing upwind flight, but little is known about the time-dependent properties of odor transduction or the mechanisms that limit receptor dynamic sensitivity. We stimulated male antennae of two moth species, Cadra cautella and Spodoptera exigua, with pheromone plumes in a wind tunnel while recording electroantennograms (EAG) and concentration of a surrogate plume (propylene, which mimics a pheromone plume) using a photoionization detector (PID). Turbulent plumes were produced by mechanical baffles, creating broad frequency range dynamic concentration changes at the antennae. Frequency response functions and coherence functions between PID and EAG signals were used to measure the dynamic responses of the two species to pheromone blends and individual components. A single time constant filter fitted the responses of both species, but S. exigua was about three times faster than C. cautella. Responses to individual pheromone components were significantly different in S. exigua but not in C. cautella. We also fitted the data with a simple block-structured nonlinear cascade. This supported the simple filter model but also suggested that the response saturates at an early stage of chemotransduction. PMID- 15537813 TI - Dopamine modulates excitability of basolateral amygdala neurons in vitro. AB - The amygdala plays a role in affective behaviors, which are modulated by the dopamine (DA) innervation of the basolateral amygdala complex (BLA). Although in vivo studies indicate that activation of DA receptors alters BLA neuronal activity, it is unclear whether DA exerts direct effects on BLA neurons or whether it acts via indirect effects on BLA afferents. Using whole cell patch clamp recordings in rat brain slices, we investigated the site and mechanisms through which DA regulates the excitability of BLA neurons. Dopamine enhanced the excitability of BLA projection neurons in response to somatic current injections via a postsynaptic effect. Dopamine D1 receptor activation increased excitability and evoked firing, whereas D2 receptor activation increased input resistance. Current- and voltage-clamp experiments in projection neurons showed that D1 receptor activation enhanced excitability by modulating a 4-aminopyridine- and alpha-dendrotoxin-sensitive, slowly inactivating K+ current. Furthermore, DA and D1 receptor activation increased evoked firing in fast-spiking BLA interneurons. Consistent with a postsynaptic modulation of interneuron excitability, DA also increased the frequency of spontaneous inhibitory postsynaptic currents recorded in projection neurons without changing release of GABA. These data demonstrate that DA exerts direct effects on BLA projection neurons and indirect actions via modulation of interneurons that may work in concert to enhance the neuronal response to large, suprathreshold inputs, while suppressing weaker inputs. PMID- 15537814 TI - Size does matter: generation of intrinsic network rhythms in thick mouse hippocampal slices. AB - Rodent hippocampal slices of < or = 0.5 mm thickness have been widely used as a convenient in vitro model since the 1970s. However, spontaneous population rhythmic activities do not consistently occur in this preparation due to limited network connectivity. To overcome this limitation, we develop a novel slice preparation of 1 mm thickness from adult mouse hippocampus by separating dentate gyrus from CA3/CA1 areas but preserving dentate-CA3-CA1 connectivity. While superfused in vitro at 32 or 37 degrees C, the thick slice exhibits robust spontaneous network rhythms of 1-4 Hz that originate from the CA3 area. Via assessing tissue O2, K+, pH, synaptic, and single-cell activities of superfused thick slices, we verify that these spontaneous rhythms are not a consequence of hypoxia and nonspecific experimental artifacts. We suggest that the thick slice contains a unitary circuitry sufficient to generate intrinsic hippocampal network rhythms and this preparation is suitable for exploring the fundamental properties and plasticity of a functionally defined hippocampal "lamella" in vitro. PMID- 15537815 TI - Progress in functional neuroanatomy: precise automatic geometric reconstruction of neuronal morphology from confocal image stacks. AB - Dendritic architecture provides the structural substrate for myriads of input and output synapses in the brain and for the integration of presynaptic inputs. Understanding mechanisms of evolution and development of neuronal shape and its respective function is thus a formidable problem in neuroscience. A fundamental prerequisite for finding answers is a precise quantitative analysis of neuronal structure in situ and in vivo. Therefore we have developed a tool set for automatic geometric reconstruction of neuronal architecture from stacks of confocal images. It provides exact midlines, diameters, surfaces, volumes, and branch point locations and allows analysis of labeled molecule distribution along neuronal surfaces as well as direct export into modeling software. We show the high accuracy of geometric reconstruction and the analysis of putative input synapse distribution throughout entire dendritic trees from in situ light microscopy preparations as a possible application. The binary version of the reconstruction module is downloadable at no cost. PMID- 15537816 TI - New type of synaptically mediated epileptiform activity independent of known glutamate and GABA receptors. AB - It is well known that excitatory synaptic transmission at the hippocampal CA3-CA1 synapse depends on the binding of released glutamate to ionotropic receptors. Here we report that during long-term application of Cs+ (5 mM), stimulation of the Schaffer collateral-commisural pathway evokes an epileptic field potential (Cs-FP) in area CA1 of the rat hippocampal slice, which is resistant to antagonists of ionotropic glutamate and GABA(A) receptors. The Cs-FP was blocked by N-type but not L-type Ca2+ channel antagonists and was attenuated by adenosine (0.5 mM), as expected for a synaptically mediated response. These properties make the Cs-FP fundamentally different from other types of Cs(+)-induced epileptiform activity. Replacement of Cs+ with antagonists of the hyperpolarization-activated nonselective cation current I(h) and inwardly rectifying potassium channels (K(IR)) or partial inhibition of the Na(+)/K+ pump did not cause Cs-FP-like potentials, which indicates that such actions of Cs+ were not responsible for the Cs-FP. The effect of Cs+ was partly mimicked by 4-aminopyridine (4-AP; 2 mM), suggesting that an increase in transmitter release is involved. The group I metabotropic glutamate receptor (mGluR) agonist (RS)-3,5-dihydroxyphenylglycine (DHPG) attenuated the Cs-FP. This effect was not, however, antagonized by group I mGluR antagonists. Selective and nonselective mGluR antagonists did not attenuate the Cs-FP. We conclude that long-term exposure to Cs+ induces a state where excitatory synaptic transmission can exist between area CA3 and CA1 in the hippocampus, independent of ionotropic and metabotropic glutamate receptors and GABA(A) receptors. PMID- 15537817 TI - Temporal dynamics of direction tuning in motion-sensitive macaque area MT. AB - We studied the temporal dynamics of motion direction sensitivity in macaque area MT using a motion reverse correlation paradigm. Stimuli consisted of a random sequence of motion steps in eight different directions. Cross-correlating the stimulus with the resulting neural activity reveals the temporal dynamics of direction selectivity. The temporal dynamics of direction selectivity at the preferred speed showed two phases along the time axis: one phase corresponding to an increase in probability for the preferred direction at short latencies and a second phase corresponding to a decrease in probability for the preferred direction at longer latencies. The strength of this biphasic behavior varied between neurons from weak to very strong and was uniformly distributed. Strong biphasic behavior suggests optimal responses for motion steps in the antipreferred direction followed by a motion step in the preferred direction. Correlating spikes to combinations of motion directions corroborates this distinction. The optimal combination for weakly biphasic cells consists of successive steps in the preferred direction, whereas for strongly biphasic cells, it is a reversal of directions. Comparing reverse correlograms to combinations of stimuli to predictions based on correlograms for individual directions revealed several nonlinear effects. Correlations for successive presentations of preferred directions were smaller than predicted, which could be explained by a static nonlinearity (saturation). Correlations to pairs of (nearly) opposite directions were larger than predicted. These results show that MT neurons are generally more responsive when sudden changes in motion directions occur, irrespective of the preferred direction of the neurons. The latter nonlinearities cannot be explained by a simple static nonlinearity at the output of the neuron, but most likely reflect network interactions. PMID- 15537818 TI - Spatial reconstruction of trajectories of an array of recording microelectrodes. AB - We present a method for estimating the locations of sites visited by an array of microelectrodes. The method relies on visualization of tracks made by electrodes coated in a fluorescent dye. These tracks are used to estimate the parameters of a simple geometrical model that generates coordinates for each recording site. We describe several ways to measure the error of this procedure and present experimental results from recordings in the motor cortex of macaque monkeys that suggest that errors are of the order of 230 microm. We also introduce a coordinate transformation that takes into account the convoluted structure of the cortex near sulci to conveniently visualize recording site locations in a rectilinear representation. This method greatly extends the capabilities of microelectrodes for studying the three-dimensional structure of topographic maps in the cortex. PMID- 15537819 TI - Distributed population mechanism for the 3-D oculomotor reference frame transformation. AB - Human saccades require a nonlinear, eye orientation-dependent reference frame transformation to transform visual codes to the motor commands for eye muscles. Primate neurophysiology suggests that this transformation is performed between the superior colliculus and brain stem burst neurons, but provides little clues as to how this is done. To understand how the brain might accomplish this, we trained a 3-layer neural net to generate accurate commands for kinematically correct 3-D saccades. The inputs to the network were a 2-D, eye-centered, topographic map of Gaussian visual receptive fields and an efference copy of eye position in 6-dimensional, push-pull "neural integrator" coordinates. The output was an eye orientation displacement command in similar coordinates appropriate to drive brain stem burst neurons. The network learned to generate accurate, kinematically correct saccades, including the eye orientation-dependent tilts in saccade motor error commands required to match saccade trajectories to their visual input. Our analysis showed that the hidden units developed complex, eye centered visual receptive fields, widely distributed fixed-vector motor commands, and "gain field"-like eye position sensitivities. The latter evoked subtle adjustments in the relative motor contributions of each hidden unit, thereby rotating the population motor vector into the correct correspondence with the visual target input for each eye orientation: a distributed population mechanism for the visuomotor reference frame transformation. These findings were robust; there was little variation across networks with between 9 and 49 hidden units. Because essentially the same observations have been reported in the visuomotor transformations of the real oculomotor system, as well as other visuomotor systems (although interpreted elsewhere in terms of other models) we suggest that the mechanism for visuomotor reference frame transformations identified here is the same solution used in the real brain. PMID- 15537820 TI - Rapid dopaminergic signaling by interneurons that contain markers for catecholamines and GABA in the feeding circuitry of Aplysia. AB - Consummatory feeding behaviors in Aplysia californica are controlled by a polymorphic central pattern generator (CPG) circuit. Previous investigations have demonstrated colocalization of markers for GABA and catecholamines within two interneurons, B20 and B65, that participate in configuring the functional output of this CPG. This study examined the contributions of GABA and dopamine (DA) to rapid synaptic signaling from B20 and B65 to follower cells that implement their specification of motor programs. Pharmacological tests did not substantiate the participation of GABA in the mediation of the excitatory postsynaptic potentials (EPSPs) from either B20 or B65. However, GABA and the GABA(B) receptor agonist baclofen were found to modify these signals in a target-specific manner. Several observations indicated that DA acts as the neurotransmitter mediating fast EPSPs from B20 to two radula closer motor neurons B8 and B16. In both motor neurons, application of DA produced depolarizing responses associated with decreased input resistance and increased excitation. B20-evoked EPSPs in both follower cells were occluded by exogenous dopamine and blocked by the DA antagonist sulpiride. While dopamine occlusion and sulpiride block of convergent signaling to B8 from B65 resembled that of B20, both of these actions were less potent on the rapid signaling from B65 to the multifunctional and widely acting interneuron B4/5. These findings indicate that dopamine mediates divergent (B20 to B16 and B8) and convergent (B20 and B65 to B8) rapid EPSPs from two influential CPG interneurons in which it is colocalized with GABA-like immunoreactivity. PMID- 15537821 TI - Glabridin, an isoflavan from licorice root, inhibits inducible nitric-oxide synthase expression and improves survival of mice in experimental model of septic shock. AB - (R)-4-(3,4-Dihydro-8,8-dimethyl)-2H,8H-benzo[1,2-b:3,4-b']dipyran-3yl)-1,3 benzenediol (glabridin), a flavonoid present in licorice extract, is known to have antimicrobial, anti-inflammatory, and cardiovascular protective activities. In the present study, we report the inhibitory effect of glabridin on nitric oxide (NO) production and inducible nitric oxide (iNOS) gene expression in murine macrophages. Glabridin attenuated lipopolysaccharide (LPS)-induced NO production in isolated mouse peritoneal macrophages and RAW 264.7 cells, a mouse macrophage like cell line. Moreover, iNOS mRNA expression was also blocked by glabridin treatment in LPS-stimulated RAW 264.7 cells. Further study demonstrated that the LPS-induced nuclear factor (NF)-kappaB/Rel DNA binding activity and NF-kappaB/Rel dependent reporter gene activity were significantly inhibited by glabridin in RAW 264.7 cells and that this effect was mediated through the inhibition of inhibitory factor-kappaB degradation and p65 nuclear translocation. Moreover, reactive oxygen species generation was also suppressed by glabridin treatment in RAW 264.7 cells. In contrast, the activity of mitogen-activated protein kinases was unaffected by glabridin treatment. In animal model, in vivo administration of glabridin increased the rate of survival of LPS-treated mice and inhibited LPS induced increase in plasma concentrations of nitrite/nitrate and tumor necrosis factor-alpha. Collectively, these data suggest that glabridin inhibits NO production and iNOS gene expression by blocking NF-kappaB/Rel activation and that this effect was mediated, at least in part, by inhibiting reactive oxygen species generation. Furthermore, in vivo anti-inflammatory effect of glabridin suggests a possible therapeutic application of this agent in inflammatory diseases. PMID- 15537822 TI - Protease inhibitor treatments reveal specific involvement of matrix metalloproteinase-9 in human adipocyte differentiation. AB - We previously showed that human and murine 3T3-F442A preadipocytes produced and released matrix metalloproteinases (MMPs) 2 and 9 and that a treatment by MMP inhibitors resulted in the blockade of murine fat cell adipose conversion. In parallel, investigators reported that other protease inhibitors, the human immunodeficiency virus (HIV) protease inhibitors (PIs) involved in lipodystrophy in humans, also reduced the adipocyte differentiation process of several murine cell lines. The present work was performed to define the effects of MMP inhibitors and HIV-PIs on the human adipocyte differentiation process, to clarify the involvement of MMPs in the control of human adipogenesis, and to determine whether HIV-PIs interact with MMPs in the control of this process. The effect of two MMP inhibitor and four HIV-PI treatments on the differentiation of primary culture human preadipocytes, as well as the putative relationships between HIV PIs and MMP-2 and -9 expression, release, or activity were investigated. We showed that MMP inhibitors and HIV-PIs reduced the human adipocyte differentiation process as assessed by the decrease of cell protein and/or triglyceride contents and expression of fatty acid binding protein and hormone sensitive lipase, two adipocyte markers. Unlike MMP inhibitors, HIV-PIs were devoid of any effect per se on recombinant MMP-2 and 9 activities but reduced the expression and release of MMP-9 by human preadipocytes. Thus, the present study indicates that the modulation of the extracellular matrix components through the production and/or activity of MMPs, and, more precisely, MMP-9 might be a key factor in the regulation of human adipose tissue development. PMID- 15537823 TI - Ligand discovery using small molecule microarrays. AB - Small molecule microarrays have recently been used to identify ligands for several proteins, and several themes regarding screening strategies and limitations have emerged. In this review, some of the technical issues related to the manufacture and screening of small molecule microarrays, as well as prospects for small molecule microarrays in several areas of drug discovery and chemistry, are discussed. PMID- 15537824 TI - Green tea polyphenol-induced epidermal keratinocyte differentiation is associated with coordinated expression of p57/KIP2 and caspase 14. AB - Epigallocatechin-3-gallate (EGCG), the most abundant polyphenol in green tea, exerts chemopreventive effects by selectively inducing apoptosis in tumor cells. In contrast, EGCG accelerates terminal differentiation in normal human epidermal keratinocytes (NHEK) mediated partially by up-regulation of p57/KIP2, a cyclin dependent kinase inhibitor that confers growth arrest and differentiation. However, it is unclear if EGCG modulates caspase 14, a unique regulator of epithelial cell terminal differentiation associated with cornification. Here, we examined the effect of EGCG on caspase 14 expression in NHEK and correlated the protein and mRNA expression of p57/KIP2 with those of caspase 14 in either normal keratinocytes or p57/KIP2-expressing tumor cells (OSC2, an oral squamous cell carcinoma cell line). Additionally, paraffin-embedded normal and untreated psoriatic (aberrant keratinization) skin sections from humans were assessed for caspase 14 by immunohistochemistry. In NHEK, EGCG induced the expression of caspase 14 mRNA and protein levels within a 24-h period. The expression of p57/KIP2 in OSC2 cells was adequate to induce caspase 14 in the absence of EGCG; this induction of caspase 14 was down-regulated by transforming growth factor beta1. In human psoriatic skin samples, caspase 14 staining in the upper epidermis was reduced, especially in nuclear areas. These results suggest that, in addition to p57/KIP2, EGCG-induced terminal differentiation of epidermal keratinocytes involves up-regulation of caspase 14. Further understanding of how EGCG modulates cellular differentiation may be useful in developing green tea preparations for selected clinical applications. PMID- 15537825 TI - Corticotropin-releasing factor receptor 1 and central heart rate regulation in mice during expression of conditioned fear. AB - The present study was performed to 1) determine heart rate (HR) effects mediated through central corticotropin-releasing factor receptor subtypes 1 (CRF(1)) investigate and 2 (CRF(2)) and 2) to the contribution of endogenous CRF to baseline HR and its fear-induced adjustment in freely moving mice. CRF ligands were injected into both lateral ventricles (i.c.v.) 15 min before the presentation of a conditioned auditory fear stimulus (CS). Initial behavioral results suggest an ovine CRF (oCRF)-mediated enhanced baseline fear and mildly enhanced conditioned auditory fear. In contrast, i.c.v. injection of oCRF (35-210 ng/mouse) dose-dependently decreased baseline HR, increased HR variability, and attenuated the CS-induced tachycardia. This effect is suggested to depend on a combined activation of sympathetic and parasympathetic activity referred to as enhanced sympathovagal antagonism. An extreme bradycardia was elicited by oCRF injection into the lower brainstem. All HR effects were probably mediated by CRF(1) because injection of the CRF(2)-selective agonist mouse urocortin II was ineffective, and the baseline bradycardia by i.c.v. CRF was preserved in CRF(2) deficient mice. Injection of various CRF receptor antagonists including the CRF(2)-selective antisauvagine-30 did not affect the conditioned HR response. This finding suggests that endogenous CRF does not contribute to the fear mediated tachycardia. Thus, the hypothesis of an involvement of CRF in HR responses of mice to acute aversive stimulation is rejected. Pharmacological evidence points at the involvement of CRF(1) in enhanced sympathovagal antagonism, a pathological state contributing to elevated cardiac risk, whereas the physiological role of the brain CRF system in cardiovascular regulation remains to be determined. PMID- 15537826 TI - Phospholipase d signaling and extracellular signal-regulated kinase-1 and -2 phosphorylation (activation) are required for maximal phorbol ester-induced transglutaminase activity, a marker of keratinocyte differentiation. AB - Protein kinase C (PKC)-activating 12-O-tetradecanoylphorbol 13-acetate (TPA) stimulates phospholipase D (PLD) activity in primary mouse epidermal keratinocytes. PLD catalyzes the hydrolysis of phosphatidylcholine to yield phosphatidic acid (PA), which can be dephosphorylated to produce PKC-activating diacylglycerol. In the presence of small amounts of a primary alcohol, PLD can instead produce novel phosphatidylalcohols at the expense of PA and diacylglycerol. Here, we have demonstrated that inhibiting PLD signal generation with 1-butanol reduced TPA-stimulated transglutaminase activity, a marker of keratinocyte differentiation. On the other hand, the structurally related tertiary alcohol tert-butanol, which cannot be used by PLD, had no effect on TPA induced transglutaminase activity. Since TPA activates all conventional and novel PKC isoforms directly, yet cannot overcome 1-butanol-mediated inhibition, this result suggests that PLD mediates its effects on transglutaminase activity (and keratinocyte differentiation) through an effector enzyme system distinct from the conventional or novel PKC isoenzymes. Data in the literature suggest that PA can recruit Raf-1 to the membrane, where it can be activated and initiate the mitogen activated protein kinase cascade that culminates in activation of extracellular signal-regulated kinase (ERK)-1 and -2. Indeed, we found that inhibition of ERK 1/2 phosphorylation (activation) inhibited TPA-induced transglutaminase activity. However, inhibition of PLD-mediated signal generation had only a small effect on TPA-elicited ERK-1/2 phosphorylation (activation), whereas inhibition of ERK-1/2 did not affect PLD activation, suggesting that these two pathways likely operate largely in parallel. Thus, our results suggest the independent involvement of the PLD and ERK-1/2 pathways in mediating transglutaminase activity and keratinocyte differentiation. PMID- 15537827 TI - Differential activation of heme oxygenase-1 by chalcones and rosolic acid in endothelial cells. AB - The induction of heme oxygenase-1 (HO-1) is widely recognized as an effective cellular strategy to counteract a variety of stressful events. We have shown that curcumin and caffeic acid phenethyl ester, two naturally occurring phytochemicals that possess antioxidant, anti-inflammatory, and anticarcinogenic activities, induce HO-1 in many cell types. This suggests that stimulation of HO-1 could partly underlie the beneficial effects exerted by these plant-derived constituents. Here we examined the ability of additional plant constituents to up regulate heme oxygenase activity and HO-1 in aortic endothelial cells. Incubation of endothelial cells with a series of polyphenolic chalcones (5-50 microM) resulted in increased heme oxygenase activity; interestingly, the chemical structure dictated the pattern of heme oxygenase induction, which was unique to each particular compound employed. We also found that rosolic acid, a constituent isolated from the rhizome of Plantago asiatica L. dramatically increased HO-1 in a concentration- and time-dependent manner. Severe cytotoxicity was observed after prolonged exposure (24 or 48 h) of cells to curcumin and caffeic acid phenethyl ester, whereas 2'-hydroxychalcone and rosolic acid did not affect cell viability. By using different mitogen-activated protein kinase inhibitors, we determined that the extracellular signal-regulated kinase, p38, and c-Jun NH(2) terminal protein kinase pathways play only a minor role in the induction of HO-1 by rosolic acid and 2'-hydroxychalcone. On the other hand, increased intra- and extracellular thiols markedly reduced the rise in heme oxygenase activity elicited by rosolic acid. Thus, this study identified novel plant constituents that highly induce HO-1 in endothelial cells and investigated some of the mechanisms involved in this effect. PMID- 15537828 TI - TRP channels at a glance. PMID- 15537829 TI - The case for nuclear translation. AB - Although it is frequently assumed that translation does not occur in eukaryotic nuclei, recent evidence suggests that some translation can take place and that it is closely coupled to transcription. The first evidence concerns the destruction of nuclear mRNAs containing premature termination codons by nonsense-mediated decay (NMD). Only ribosomes can detect termination codons, and as some NMD occurs within the nuclear fraction, active nuclear ribosomes could perform the required detection. The second evidence is the demonstration that tagged amino acids are incorporated into nascent polypeptides in a nuclear process coupled to transcription. The third evidence is that components involved in translation, NMD and transcription colocalize, coimmunoprecipitate and co-purify. All these results are simply explained if nuclear ribosomes scan nascent transcripts for premature termination codons at the site of transcription. Alternatively, the scanning needed for NMD might take place at the nuclear membrane, and contaminating cytoplasmic ribosomes might give the appearance of some nuclear translation. We argue, however, that the balance of evidence favours bona fide nuclear translation. PMID- 15537830 TI - Tau phosphorylation in neuronal cell function and dysfunction. AB - Tau is a group of neuronal microtubule-associated proteins that are formed by alternative mRNA splicing and accumulate in neurofibrillary tangles in Alzheimer's disease (AD) brain. Tau plays a key role in regulating microtubule dynamics, axonal transport and neurite outgrowth, and all these functions of tau are modulated by site-specific phosphorylation. There is significant evidence that a disruption of normal phosphorylation events results in tau dysfunction in neurodegenerative diseases, such as AD, and is a contributing factor to the pathogenic processes. Indeed, the abnormal tau phosphorylation that occurs in neurodegenerative conditions not only results in a toxic loss of function (e.g. decreased microtubule binding) but probably also a toxic gain of function (e.g. increased tau-tau interactions). Although tau is phosphorylated in vitro by numerous protein kinases, how many of these actually phosphorylate tau in vivo is unclear. Identification of the protein kinases that phosphorylate tau in vivo in both physiological and pathological processes could provide potential therapeutic targets for the treatment of AD and other neurodegenerative diseases in which there is tau pathology. PMID- 15537831 TI - Inducible changes in cell size and attachment area due to expression of a mutant SWI/SNF chromatin remodeling enzyme. AB - The SWI/SNF enzymes belong to a family of ATP-dependent chromatin remodeling enzymes that have been functionally implicated in gene regulation, development, differentiation and oncogenesis. BRG1, the catalytic core subunit of some of the SWI/SNF enzymes, can interact with known tumor suppressor proteins and can act as a tumor suppressor itself. We report that cells that inducibly express ATPase deficient versions of BRG1 increase in cell volume, area of attachment and nuclear size upon expression of the mutant BRG1 protein. Examination of focal adhesions reveals qualitative changes in paxillin distribution but no difference in the actin cytoskeletal structure. Increases in cell size and shape correlate with over-expression of two integrins and the urokinase-type plasminogen activator receptor (uPAR), which is also involved in cell adhesion and is often over-expressed in metastatic cancer cells. These findings demonstrate that gene expression pathways affected by chromatin remodeling enzymes can regulate the physical dimensions of mammalian cell morphology. PMID- 15537832 TI - Alteration of chromosome positioning during adipocyte differentiation. AB - Chromosomes are highly restricted to specific chromosome territories within the interphase nucleus. The arrangement of chromosome territories is non-random, exhibiting a defined radial distribution as well as a preferential association with specific nuclear compartments, which indicates a functional role for chromosome-territory organization in the regulation of gene expression. In this report, we focus on changes in adipocyte differentiation that are related to a specific chromosomal translocation associated with liposarcoma tumorigenesis, t(12;16). We have examined the relative and radial positioning of the chromosome territories of human chromosomes 12 and 16 during adipocyte differentiation, and detected a close association between the territories of chromosomes 12 and 16 in differentiated adipocytes, an association not observed in preadipocytes. Although further studies are required to elucidate the underlying reasons for the adipocyte-specific translocation of chromosomes 12 and 16, our observations indicate that alteration of relative chromosome positioning might play a key role in the tumorigenesis of human liposarcomas. In addition, these results demonstrate the potential impact of higher order chromatin organization on the epigenetic mechanisms that control gene expression and gene silencing during cell differentiation. PMID- 15537833 TI - Functional significance of a natural allelic variant of human carbonyl reductase 3 (CBR3). AB - Human carbonyl reductase (CBR) activity accounts for a significant fraction of the metabolism of endogenous and xenobiotic carbonyl compounds. It is possible that genetic polymorphisms in CBR1 and CBR3 are key for the wide interindividual variability in the disposition of CBR drug substrates. We pinpointed a single nucleotide polymorphism in CBR3 (CBR3 V244M) that encodes for a V244 to M244 change. Blacks showed a higher frequency of the M244 allele (q = 0.51, n = 49) than did whites (q = 0.31, n = 70; p = 0.003). In addition, DNA variation panels from 10 ethnic groups presented a wide range of CBR3 V244M genotype distributions. Kinetic experiments with the recombinant CBR3 protein variants and menadione revealed that CBR3 M244 has significantly higher V(max) than does CBR3 V244 (V(max) CBR3 M244 = 40.6 +/- 1.3 micromol/min x mg versus V(max) CBR3 V244 = 19.6 +/- 2.0 micromol/min x mg, p = 0.002). In contrast, both isoforms presented similar K(m) values (K(m) CBR3 M244 = 22.9 +/- 2.9 microM versus K(m) CBR3 V244 = 24.6 +/- 3.2 microM, p = 0.43). Assays with NADP(H) demonstrated a higher V(maxNADP(H)) (1.6-fold) and increased catalytic efficiency (V(maxNADP(H))/K(mNADP(H))) for CBR3 M244 compared with CBR3 V244 (p = 0.013). Comparative three-dimensional analyses based on the structure of the homologous porcine carbonyl reductase suggested that the V244M substitution is positioned in a region critical for interactions with the NADP(H) cofactor. These studies demonstrate that the common CBR3 V244M polymorphism encodes for CBR3 isoforms with distinctive enzymatic properties. PMID- 15537834 TI - Lansoprazole enantiomer activates human liver microsomal CYP2C9 catalytic activity in a stereospecific and substrate-specific manner. AB - We recently proposed a possible stereoselective activation by lansoprazole of CYP2C9-catalyzed tolbutamide hydroxylation, as well as stereoselective inhibition of several cytochrome P450 (P450) isoforms. This study evaluated the effects of lansoprazole enantiomers on CYP2C9 activity in vitro, using several probe substrates. For tolbutamide 4-methylhydroxylation and phenytoin 4-hydroxylation, R-lansoprazole was an activator (140 and 550% of control at 100 microM R lansoprazole, EC50 values of 19.9 and 30.2 microM, respectively). R-Lansoprazole mediated activation of the formation of 4-hydroxyphenytoin was also seen with recombinant human CYP2C9. R-Lansoprazole increased the Michaelis-Menten-derived V(max) of phenytoin 4-hydroxylation from 0.024 to 0.121 pmol/min/pmol P450, and lowered its K(m) from 20.5 to 15.0 microM, suggesting that R-lansoprazole activates CYP2C9-mediated phenytoin metabolism without displacing phenytoin from the active site. Kinetic parameters were also estimated using the two-site binding equation, with alpha values <1 and beta values >1, indicative of activation. Additionally, phenytoin at 10 to 200 microM had no reciprocal effect on the hydroxylation of R-lansoprazole. Meanwhile, R-lansoprazole had no activation effect on diclofenac and S-warfarin metabolism in the incubation study using both recombinant CYP2C9 and human liver microsomes. These substrate dependent activation effects suggest that phenytoin has a different binding orientation compared with diclofenac and S-warfarin. Overall, these results suggest that R-lansoprazole activates CYP2C9 in a stereospecific and substrate specific manner, possibly by binding within the active site and inducing positive cooperativity. This is the first report to describe stereoselective activation of this cytochrome P450 isoform. PMID- 15537835 TI - A method to monitor replication fork progression in mammalian cells: nucleotide excision repair enhances and homologous recombination delays elongation along damaged DNA. AB - The capacity to rescue stalled replication forks (RFs) is important for the maintenance of cell viability and genome integrity. Here, we have developed a novel method for monitoring RF progression and the influence of DNA lesions on this process. The method is based on the principle that each RF is expected to be associated with a pair of single-stranded ends, which can be analyzed by employing strand separation in alkali. This method was applied to examine the rate of RF progression in Chinese hamster cell lines deficient in ERCC1, which is involved in nucleotide excision repair (NER), or in XRCC3, which participates in homologous recombination repair, following irradiation with ultraviolet (UV) light or exposure to benzo(a)pyrene-7,8-diol-9,10-epoxide (BPDE). The endpoints observed were cell survival, NER activity, formation of double-strand breaks and the rate of RF progression. Subsequently, we attempted to explain our observation that cells deficient in XRCC3 (irs1SF) exhibit enhanced sensitivity to UV radiation and BPDE. irs1SF cells demonstrated a capacity for NER that was comparable with wild-type AA8 cells, but the rate of RF progression was even higher than that for the wild-type AA8 cells. As expected, cells deficient in ERCC1 (UV4) showed no NER activity and were hypersensitive to both UV radiation and BPDE. The observation that cells deficient in NER displayed a pronounced delay in RF progression indicates that NER plays an important role in maintaining fork progression along damaged DNA. The elevated rate of RF progression in XRCC3 deficient cells indicates that this protein is involved in a time-consuming process which resolves stalled RFs. PMID- 15537836 TI - Combinatorial codon-based amino acid substitutions. AB - Twenty Fmoc-protected trinucleotide phosphoramidites representing a complete set of codons for the natural amino acids were chemically synthesized for the first time. A pool of these reagents was incorporated into oligonucleotides at substoichiometric levels to generate two libraries of variants that randomly carry either few or many codon replacements on a region encoding nine amino acids of the bacterial enzyme TEM-1 beta-lactamase. Assembly of the libraries was performed in a completely automated mode through a simple modification of ordinary protocols. This technology eliminates codon redundancy, stop codons and enables complete exploration of sequence space for single, double and triple mutations throughout a protein region spanning several residues. Sequence analysis of many non-selected clones revealed a good incorporation of the trinucleotides, producing combinations of mutations quite different from those obtained using conventional degenerate oligonucleotides. Ceftazidime-selection experiments yielded several never before reported variants containing novel amino acid combinations in the beta-lactamase omega loop region. PMID- 15537837 TI - X-ray photoelectron spectroscopy and infrared spectroscopy study of maleimide activated supports for immobilization of oligodeoxyribonucleotides. AB - Surface-tethered nucleic acids are widely applied in solid-phase assays in which complementary strands must be detected against a complex mixture of other sequences. In response to such needs, numerous methods have been developed for immobilizing nucleic acids on solid supports. Often, detailed analysis of associated chemical transformations and of potential side reactions is difficult to obtain. Combined use of planar and high surface area powder supports allows characterization using surface as well as bulk diagnostic techniques. This approach is followed in the present study in which X-ray photoelectron spectroscopy (XPS), transmission infrared spectroscopy (FTIR) and reactivity titrations are used to investigate siliceous supports modified with an aminosilane precursor followed by a maleimide-bearing crosslinker for attachment of nucleic acids. The supports retain maleimide activity for approximately a day when stored under buffer, but deactivation is accelerated under basic conditions or by incomplete conversion of the precursor aminosilane monolayer. Reactions involving the olefinic bond of the imide as well as its carbonyl groups are observed and analyzed. Attachment of sulfhydryl-terminated oligodeoxyribonucleotides is highly site specific, and immobilized strands exhibit excellent hybridization activity. Quantitative use of XPS for label-free determination of DNA coverage based on calibration against reference materials is also described. PMID- 15537838 TI - Renal cell carcinoma: analysis of postoperative recurrence patterns. AB - PURPOSE: To retrospectively analyze the recurrence patterns of renal cell carcinoma (RCC) and the factors affecting tumor recurrence. MATERIALS AND METHODS: The institutional review board approved this study; patient informed consent was not required. There were 162 men (mean age, 54 years +/- 13 [standard deviation]) and 32 women (mean age, 56 years +/- 11) who had undergone complete surgical resection of RCC. Mean follow-up period was 45 months (range, 7-92 months). In consensus, two radiologists determined the presence or absence of tumor recurrence and recorded the time and sites of tumor recurrence. The relationships between tumor recurrence and tumor factors, including greatest diameter (> or =5 cm or <5 cm), T stage, N stage, stage group, histologic subtype, and nuclear grade, were evaluated by using Kaplan-Meier statistics. RESULTS: Tumor recurred in 41 (21%) patients. The mean time of tumor recurrence was 17 months (range, 3-50 months). Tumor recurred within 2 years after surgery in 34 (83%) patients. Tumor recurrence sites included lung (n = 29), bone (n = 13), the nephrectomy site (n = 7), brain (n = 6), liver (n = 5), mediastinal lymph nodes (n = 5), the contralateral kidney (n = 4), and the neck muscles (n = 2). The recurrence rate was greater for tumors 5 cm or larger than for those smaller than 5 cm, greater for T3a or T3b tumors than for T1 tumors, greater for stage III tumors than for stage I tumors, and greater for tumors with a nuclear grade of 3 or 4 than for those with a nuclear grade of 1 or 2 (P < .05 for all). CONCLUSION: RCC usually recurs within 2 years after surgery, with the lung being the most vulnerable site; greatest tumor diameter, T stage, stage group, and nuclear grade are important factors for recurrence. PMID- 15537839 TI - Pulmonary nodules on multi-detector row CT scans: performance comparison of radiologists and computer-aided detection. AB - PURPOSE: To compare the performance of radiologists and of a computer-aided detection (CAD) algorithm for pulmonary nodule detection on thin-section thoracic computed tomographic (CT) scans. MATERIALS AND METHODS: The study was approved by the institutional review board. The requirement of informed consent was waived. Twenty outpatients (age range, 15-91 years; mean, 64 years) were examined with chest CT (multi-detector row scanner, four detector rows, 1.25-mm section thickness, and 0.6-mm interval) for pulmonary nodules. Three radiologists independently analyzed CT scans, recorded the locus of each nodule candidate, and assigned each a confidence score. A CAD algorithm with parameters chosen by using cross validation was applied to the 20 scans. The reference standard was established by two experienced thoracic radiologists in consensus, with blind review of all nodule candidates and free search for additional nodules at a dedicated workstation for three-dimensional image analysis. True-positive (TP) and false-positive (FP) results and confidence levels were used to generate free response receiver operating characteristic (ROC) plots. Double-reading performance was determined on the basis of TP detections by either reader. RESULTS: The 20 scans showed 195 noncalcified nodules with a diameter of 3 mm or more (reference reading). Area under the alternative free-response ROC curve was 0.54, 0.48, 0.55, and 0.36 for CAD and readers 1-3, respectively. Differences between reader 3 and CAD and between readers 2 and 3 were significant (P < .05); those between CAD and readers 1 and 2 were not significant. Mean sensitivity for individual readings was 50% (range, 41%-60%); double reading resulted in increase to 63% (range, 56%-67%). With CAD used at a threshold allowing only three FP detections per CT scan, mean sensitivity was increased to 76% (range, 73%-78%). CAD complemented individual readers by detecting additional nodules more effectively than did a second reader; CAD-reader weighted kappa values were significantly lower than reader-reader weighted kappa values (Wilcoxon rank sum test, P < .05). CONCLUSION: With CAD used at a level allowing only three FP detections per CT scan, sensitivity was substantially higher than with conventional double reading. PMID- 15537840 TI - Out of whack, out of mind. Misaligned brain cells might sap memory as we age. PMID- 15537841 TI - Proteomics in aging-related research. AB - Aging is characterized by changes in a variety of cellular phenotypes, which in turn are caused by alterations in the concentrations, compositions, and interactions of cellular proteins. The characterization of these age-associated protein alterations and of their effect on the ability of the organism to maintain homeostasis is thus of critical importance. Proteomics is an emerging scientific discipline that focuses on the determination and analysis of the entire protein complement of a cell (termed the proteome) and of its distribution within the cell. Proteomic analysis provides information about the levels of protein expression under a variety of conditions and about the nature of posttranscriptional modifications, and reveals specific protein complexes. By comparing the proteomes of young and old cells under a variety of conditions, detailed information on the molecular basis of their different phenotypes can be obtained. Although still under development, this scientific discipline holds great promise for aging-related research. PMID- 15537845 TI - Calcified splenic artery. PMID- 15537844 TI - Recent advances in diabetic nephropathy. AB - Diabetic nephropathy is the leading cause of end stage renal disease worldwide and is associated with increased cardiovascular risk. The earliest clinical manifestation is of microalbuminuria. Tight blood glucose and blood pressure control reduce the risk of microalbuminuria. Once microalbuminuria is present, the rate of progression to end stage renal disease and of cardiovascular disease can be delayed by aggressive management of blood pressure, glucose, and lipids. Inhibition of the renin-angiotensin system is important to reduce intraglomerular pressure but other classes of antihypertensive agent may also be needed to gain adequate control of systemic blood pressure. Such measures can at least half the rate of progression of nephropathy and cardiovascular disease. PMID- 15537846 TI - Management of portal hypertension. AB - Treatment of portal hypertension is evolving based on randomised controlled trials. In acute variceal bleeding, prophylactic antibiotics are mandatory, reducing mortality as well as preventing infections. Terlipressin or somatostatin combined with endoscopic ligation or sclerotherapy is the best strategy for control of bleeding but there is no added effect of vasoactive drugs on mortality. Non-selective beta-blockers are the first choice therapy for both secondary and primary prevention; if contraindications or intolerance to beta blockers are present then band ligation should be used. Novel therapies target the increased intrahepatic resistance caused by microcirculatory intrahepatic deficiency of nitric oxide and contraction of activated intrahepatic stellate cells. PMID- 15537847 TI - Cachexia in malaria and heart failure: therapeutic considerations in clinical practice. AB - Cachexia is an independent prognostic marker of survival in many chronic diseases including heart failure and malaria. Morbidity and mortality from malaria is high in most of the third world where it presents a very challenging public health problem. Malaria may present in the UK as fever in the returning traveller or as fever in overseas visitors. How and why cachexia develops in malaria in a manner similar to the cachexia of chronic heart failure and the treatment strategies that would alter outcomes in both diseases are discussed in this review. PMID- 15537849 TI - Management of traumatic brain injury: some current evidence and applications. AB - Traumatic brain injury remains a worldwide problem. Newer modalities in the management of such injuries include both drugs and therapeutic strategies. Continuing research in animal models has provided a better understanding of the pathophysiological processes that follow head injury, and this in turn has enabled workers to work on improved treatment targets. Although there are exciting and novel approaches emerging, there is no substitute for meticulous initial resuscitation. Additionally, some of the more well known management options are now better understood. These concepts are discussed in the article. PMID- 15537851 TI - Avoiding laboratory pitfalls in infectious diseases. AB - In today's medical care environment, clinicians are challenged to order clinically relevant, cost effective laboratory tests and antibiotic therapy. Together, physicians and laboratories must have guidelines and strategies that can provide quality patient care, while minimising costs and preventing further emergence of antimicrobial drug resistance. Five clinical vignettes that demonstrate these principles are presented. PMID- 15537852 TI - Malaria in the UK: past, present, and future. AB - There is strong evidence that malaria was once indigenous to the UK, that global warming is occurring, and that human activity is contributing to global warming. Global warming will have a variety of effects, one of which will probably be the return of indigenous malaria. PMID- 15537850 TI - Methadone: applied pharmacology and use as adjunctive treatment in chronic pain. AB - This article reviews the unique pharmacological properties of methadone and outlines its appropriate clinical application, with focus upon its use in the treatment of chronic pain. Although methadone is most widely known for its use in the treatment of opioid dependence, methadone also provides effective analgesia. Patients who experience inadequate pain relief or intolerable side effects with other opioids or who suffer from neuropathic pain may benefit from a transition to methadone as their analgesic agent. Adverse effects, particularly respiratory depression and death, make a fundamental knowledge of methadone's pharmacological properties essential to the provider considering methadone as analgesic therapy for a patient with chronic pain. PMID- 15537853 TI - Anatomical distribution of colorectal cancer over a 10 year period in a district general hospital: is there a true "rightward shift"? AB - INTRODUCTION: Recent studies of patients with colorectal cancer have suggested a shift towards the proximal colon and an increase in the incidence of right sided colon cancer. This study aimed to determine the anatomical distribution of colorectal cancer over a 10 year period in a district general hospital. METHODS: Records of patients diagnosed with primary colorectal cancer from 1993 to 2002 were reviewed for demographic data, histology subtype, and anatomical location of the tumour. Tumours located at and proximal to the splenic flexure were defined as right sided cancer and tumours arising distal to the splenic flexure were defined as left sided cancer. RESULTS: A total of 763 patients were included in the study, of whom all had adenocarcinoma and 99% were white. Sixty nine percent of cancers were left sided and 31% were right sided. Although there was a 4% increase in the proportion of right sided cancers, there was no statistically significant increase using logistic regression analysis. Mann-Whitney U test revealed no significant difference in age at diagnosis between the right and left sided cancers. Although a higher proportion of females were diagnosed with right sided cancer compared with left sided cancer, this was not statistically significant. CONCLUSION: The anatomical distribution of colorectal cancer has been fairly stable at this hospital with no evidence of a shift towards the proximal colon. No differences were identified in the tumour distribution with respect to gender and age at diagnosis. Our findings support the initial application of flexible sigmoidoscopy for investigating patients with suspected colorectal malignancy and follow up colonoscopy for selected patients to exclude right sided pathology. PMID- 15537854 TI - Presentation and outcome of rhino-orbital-cerebral mucormycosis in patients with diabetes. AB - AIM: To report presentation and outcome of rhino-orbital-cerebral mucormycosis (ROCM) exclusively in patients with diabetes mellitus. METHODS: Retrospective, non-comparative, interventional analysis of the medical records of 35 patients with ROCM among 22 316 patients with diabetes seen over the last 12 years. RESULTS: A cohort of 23 men and 12 women with a mean (SD) age of 47.3 (14.4) years (range 18-70 years) was studied. Five patients had type 1 diabetes mellitus, 29 had type 2 diabetes mellitus, and one had secondary diabetes. Nine patients had ROCM as the first clinical manifestation of diabetes. The mean (SD) blood glucose at presentation was 20.6 (8.3) mmol/l (range 10.0 to 53.3 mmol/l) and 17 patients had ketosis/ketoacidosis. Ophthalmic symptoms and signs were pronounced: external ophthalmoplegia (89%), proptosis (83%), visual loss (80%), chemosis (74%), and eye lid gangrene (14%). Non-ophthalmic manifestations included sinusitis (100%), nasal discharge/ulceration (74%), infranuclear VI nerve palsy (46%), palatal necrosis (29%), cerebral lobe involvement (20%), and hemiparesis (17%). Computed tomography/magnetic resonance imaging showed involvement of paranasal sinuses in all patients with ethmoid (86%) and maxillary (80%) sinuses being most frequently involved. Orbital involvement was observed in 80% of patients with cavernous sinus thrombosis in 11%, and internal carotid occlusion and hydrocephalus in 3% each. All were treated with amphotericin B (3 3.5 g) and 26 (74%) patients underwent appropriate surgery. Twenty one patients (68%) survived with a mean (SD) follow up of 39.6 (34.1) months (range 10 months to 11 years). Factors related to poor survival included delay in diagnosis and treatment (p<0.05), facial and/or eye lid gangrene (p<0.05), hemiplegia (p<0.05), cerebral invasion by mucorales (p<0.05), and treatment with amphotericin B alone (p<0.05). CONCLUSIONS: In patients with diabetes and ROCM, ROCM was the presenting manifestation in one fourth of the patients. Ophthalmic and extensive cerebral involvement predominated in the clinical picture. Delay in treatment due to late presentation and associated complications were major determinants of the survival outcome in these patients. PMID- 15537855 TI - Pre-aortic paraganglioma. PMID- 15537856 TI - Do post-take ward round proformas improve communication and influence quality of patient care? AB - The post-take ward round is a critical time for reviewing the initial history, examination and results, and the stage at which further treatment and investigations will be determined. However documentation of this ward round is often inadequate, so the benefits of decision making are lost. The documentation of 95 ward rounds was assessed for key items of information before and after the introduction of a proforma sheet. The introduction of the proforma led to a significant improvement in the documentation of a diagnosis, management plan, prophylaxis for deep vein thrombosis, and resuscitation status (p<0.05), which will have a significant impact on patient care. PMID- 15537857 TI - Older people's understanding of cumulative risks when provided with annual stroke risk information. AB - Understanding how cumulative health risks increase over time constitutes an important part of decision making about treatment risks and benefits. In this study, wide ranging perceptions of cumulative risk led to significant overestimation and underestimation of risks over time. Time has shown to significantly impact on the personal interpretation of cumulative risk information by patients, with many subjects struggling to understand the concept of cumulative risks. This suggests that a careful explanation of the effect of cumulative risk over time to patients should contribute to more effective decision making. PMID- 15537858 TI - Symptomatic bradycardia and postural hypotension. PMID- 15537859 TI - Shortness of breath. PMID- 15537860 TI - Maternal factors controlling blastomere fragmentation in early mouse embryos. AB - Interactions between sperm and egg are required to maintain embryo viability and cellular integrity. Differential transcriptional activities and epigenetic differences that include genomic imprinting provide mechanisms by which complementary parental genome functions support early embryogenesis. We previously showed that cytofragmentation can be influenced by the specific combination of maternal and paternal genotypes. Using maternal pronuclear transfer in mouse embryos, we examined the cellular basis for the maternal genotype effect. We found that the maternal genotype effect is predominantly controlled by the maternal pronucleus, with a lesser role played by the ooplasm. This effect of the maternal pronucleus is sensitive to alpha-amanitin treatment. The effect of the maternal component of the embryonic genome on cytofragmentation constitutes the earliest known effect of the embryonic genome on mammalian embryo phenotype. The results also indicate that clinical procedures seeking to define or manipulate oocyte quality in humans should take into account early effects of the embryonic genome, particularly the maternal genome. PMID- 15537861 TI - Sperm mobility: phenotype in roosters (Gallus domesticus) determined by mitochondrial function. AB - Previously, inheritance of sperm mobility entailed a maternal additive genetic effect, and sperm ATP content was correlated (r = 0.80) with phenotype. The present study was conducted to determine if mitochondrial function was critical to phenotypic expression. Whereas phenotype was independent of mitochondrial helix length, phenotype was correlated with sperm oxygen consumption (r = 0.83) using random-bred roosters. Aberrant mitochondria characterized immobile sperm, as evidenced by transmission-electron microscopy. Such mitochondria were swollen and contained disorganized cristae. Additional experiments were performed with roosters from lines selected for low or high sperm mobility. A threefold difference in sperm oxygen consumption was observed between lines. Single nucleotide polymorphisms were observed in mitochondrial DNA by sequencing replicate mitochondrial genomes from each line. An A-to-G substitution in the gene encoding tRNA(Arg) was inherited consistently, as evidenced by restriction fragment length polymorphism analysis using two male and two female progeny per family group and 14 family groups per line. Motile concentration in semen from low-line males was half that observed in semen from high-line males, as evidenced by computer-assisted sperm motion analysis. Likewise, 47% of sperm from low-line males contained aberrant mitochondria, compared to 4% for high-line males. In summary, sperm mobility phenotype was dependent on mitochondrial function, which in turn was altered by genetic selection. Fowl deferent duct fluid contains a high concentration of glutamate. We propose that variation in sperm mobility phenotype stems from the extent to which glutamate induces excessive mitochondrial Ca2+ uptake before ejaculation. PMID- 15537862 TI - Identification of the leukemia inhibitory factor cell targets within the rat testis. AB - Leukemia inhibitory factor (LIF), a pleiotropic cytokine, is expressed in the rat testis and produced predominantly by peritubular myoid cells. The aims of this study were to characterize the testicular cell targets of LIF and to identify the role of LIF in the testis. The LIF receptor (LIF-R)/gp190 transcript was detected by reverse transcription-polymerase chain reaction (RT-PCR) in the rat testis from Day 13.5 postcoitum until adulthood. Seven highly purified testicular cell populations, representative of the major testicular constituents, were studied at transcriptional and protein levels by, respectively, RT-PCR and flow cytometry with biotinylated-LIF. Spermatogonia and, to a lesser extent, the somatic cells, exhibited specific LIF-binding sites. These results were strengthened by in situ analysis, showing predominant LIF-R immunoreactivity in spermatogonia at all ages studied. In addition to the 190-kDa LIF-R, Western blot analysis revealed the presence of a 50- to 60-kDa C-terminal gp190 isoform. This truncated form, which is unable to bind LIF, was the only form expressed in meiotic germ cells, suggesting an original down-regulation process of LIF-R expression during spermatogenesis. Finally, we showed that LIF increased [3H]-thymidine incorporation in spermatogonia in microdissected, cultured seminiferous tubules. Taken together, our results strongly suggest that LIF has a role in the regulation of the spermatogonial cell compartment. PMID- 15537863 TI - Developmental sensitivity of the bovine corpus luteum to prostaglandin F2alpha (PGF2alpha) and endothelin-1 (ET-1): is ET-1 a mediator of the luteolytic actions of PGF2alpha or a tonic inhibitor of progesterone secretion? AB - We examined the responsiveness of large luteal cells (LLC), small luteal cells (SLC), and endothelial cells of the Day 4 and Day 10 bovine corpus luteum (CL) to prostaglandin (PG) F2alpha and endothelin (ET)-1. Using a single-cell approach, we tested the ability of each agonist to increase the cytoplasmic concentration of calcium ions ([Ca2+]i) as function of luteal development. All tested concentrations of agonists significantly (P = 0.05) increased [Ca2+]i in all cell populations isolated from Day 4 and Day 10 CL. Day 10 steroidogenic cells were more responsive than Day 4 cells to PGF2alpha and ET-1. Response amplitudes and number of responding cells were affected significantly by agonist concentration, luteal development, and cell type. Response amplitudes were greater in LLC than in SLC; responses of maximal amplitude were elicited with lower agonist concentrations in Day 10 cells than in Day 4 cells. Furthermore, on Day 10, as the concentration of PGF2alpha increased, larger percentages of SLC responded. Endothelial cells responded maximally, regardless of agonist concentration and luteal development. In experiment 2, we tested the developmental responsiveness of total dispersed and steroidogenic-enriched cells to the inhibitory actions of PGF2alpha and ET-1 on basal and LH-stimulated progesterone accumulation. The potency of PGF2alpha steroidogenic-enriched cells on Day 4 was lower than on Day 10; in contrast, the potency of ET-1 was not different. Therefore, ET-1 was a tonic inhibitor of progesterone accumulation rather than a mediator of PGF2alpha action. The lower efficacy of PGF2alpha in the early CL more likely is related to signal transduction differences associated with its receptor at these two developmental stages than to the inability of PGF2alpha to up-regulate ET-1. PMID- 15537864 TI - Ultrastructural and immunohistochemical characterization of the bovine epiblast. AB - The epiblast represents the final embryonic founder cell population with the potential for giving rise to all cell types of the adult body. The pluripotency of the epiblast is lost during the process of gastrulation. Large animal species have a lack of specific markers for pluripotency. The aim of the present study was to characterize the bovine epiblast cell population and to provide such markers. Bovine Day 12 and Day 14 embryos were processed for transmission electron microscopy or immunohistochemistry. In Day 12 embryos, two cell populations of the epiblast were identified: one constituting a distinctive basal layer apposing the hypoblast, and one arranged inside or above the former layer, including cells apposing the Rauber layer. Immunohistochemically, staining for the octamer-binding transcription factor 4 (OCT4, also known as POU5F1), revealed a specific and exclusive staining of nuclei of the complete epiblast. Colocalization of vimentin and OCT4 was demonstrated. Only trophectodermal cells stained for alkaline phosphatase. Staining for the proliferation marker Ki-67 was localized to most nuclei throughout the epiblast. A continuous staining for zonula occludens-1 protein was found between cells of the trophectoderm and hypoblast but was not evident in the epiblast. A basement membrane, detected by staining for laminin, formed a "cup-like" structure in which the epiblast was located. The ventrolateral sides of the cup appeared to be incomplete. In conclusion, the bovine epiblast includes at least two cell subpopulations, and OCT4 was shown, to our knowledge for the first time, to be localized exclusively to epiblast cells in this species. PMID- 15537865 TI - Regulation of follicle-stimulating hormone-receptor messenger RNA in hen granulosa cells relative to follicle selection. AB - Both the viability of hen prehierarchal follicles and subsequent differentiation associated with the selection of a single follicle per day into the preovulatory hierarchy depend on circulating FSH and the expression of FSH receptor (FSH-R) in granulosa cells. The present study addresses mechanisms that mediate both basal expression plus selective up-regulation of FSH-R mRNA in granulosa cells from prehierarchal follicles. Results demonstrate that FSH-R mRNA is both expressed and functional in granulosa cells collected from growing prehierarchal follicles as small as those of 1-2 mm in diameter, as indicated by rapid induction of steroidogenic acute regulatory (StAR) protein expression by FSH in vitro. Real time polymerase chain reaction determined that relative FSH-R expression within the granulosa layer from individual prehierarchal follicles of 6-8 mm in diameter was similar among the 8-13 follicles within this cohort, with the notable exception that the granulosa layer from a single follicle (presumably the selected follicle) showed elevated expression. Levels of FSH-R mRNA expression were enhanced by both recombinant human (rh) transforming growth factor (TGF) beta1 and, to a lesser extent, rh-activin A after 20 h of culture. This stimulatory effect was effectively blocked by mitogen-activated protein (MAP) kinase signaling induced by TGF alpha treatment. Finally, inhibition of MAP kinase signaling, using the selective inhibitor U0126, promoted FSH-R expression and further enhanced TGF beta1-induced FSH-R expression in vitro. Collectively, results suggest that premature granulosa cell differentiation normally is suppressed by tonic MAP kinase signaling. At the time of follicle selection, a release from inhibitory MAP kinase signaling is proposed to occur, which enables the full potentiation of FSH-R expression mediated by intrafollicular factors. PMID- 15537866 TI - A model for implantation: coculture of blastocysts and uterine endometrium in mice. AB - One of the limitations in embryo implantation research is the lack of an available in vitro model that faithfully replicates embryo-uterine interactions. In previous studies, embryos were cultured on a monolayer of either uterine epithelial cells or extracellular matrix substratum on which embryos could adhere and outgrow. However, these models failed to display embryonic invasion, primarily because of the shortage of critical structural and molecular supports that are available in vivo. In the present study, we used intact mouse uterine endometrium collected on Day 4 of pregnancy and placed in contact with blastocysts to initiate coculture experiments in a defined medium at the air liquid interface. The culture medium was composed of Ham F-12/Dulbecco modified Eagle medium (1:1), 30% fetal calf serum, 63.5 nmol/L of progesterone, 7.14 nmol/L of estradiol-17beta, 100 mug/ml of insulin, and 20 ng/ml of epidermal growth factor, whereas the incubation condition was mixed air of 50% oxygen, 5% carbon dioxide, and 45% nitrogen with a humidity of greater than 90% at 37 degrees C. Our observations from 24 h of culture clearly demonstrated that embryos were capable of attachment to the uterine endometrium and displayed partial invasion into the endometrial stroma. Interestingly, no outgrowth of trophoblasts on the surface of uterine endometrium was seen, but embryos exhibited a pole-specific attachment. Overall, this model is capable of demonstrating a true invasion of embryo within the endometrial stroma and may be suitable in studies related to early embryo implantation. PMID- 15537867 TI - Transport of bile acids, sulfated steroids, estradiol 17-beta-D-glucuronide, and leukotriene C4 by human multidrug resistance protein 8 (ABCC11). AB - We previously determined that expression of human multidrug resistance protein (MRP) 8, a recently described member of the MRP family of ATP-binding cassette transporters, enhances cellular extrusion of cyclic nucleotides and confers resistance to nucleotide analogs (J Biol Chem 278:29509-29514, 2003). However, the in vitro transport characteristics of the pump have not been determined. In this study, the substrate selectivity and biochemical activity of MRP8 is investigated using membrane vesicles prepared from LLC-PK1 cells transfected with MRP8 expression vector. Expression of MRP8 is shown to stimulate the ATP dependent uptake of a range of physiological and synthetic lipophilic anions, including the glutathione S-conjugates leukotriene C4 and dinitrophenyl S glutathione, steroid sulfates such as dehydroepiandrosterone 3-sulfate (DHEAS) and estrone 3-sulfate, glucuronides such as estradiol 17-beta-D-glucuronide (E(2)17betaG), the monoanionic bile acids glycocholate and taurocholate, and methotrexate. In addition, MRP8 is competent in the in vitro transport of cAMP and cGMP, in accord with the results of our previously reported cellular studies. DHEAS, E(2)17betaG, and methotrexate were transported with K(m) and V(max) values of 13.0 +/- 0.8 microM and 34.9 +/- 9.5 pmol/mg/min, 62.9 +/- 12 microM and 62.0 +/- 5.2 pmol/mg/min, and 957 +/- 28 microM and 317 +/- 17 pmol/mg/min, respectively. Based upon the stimulatory action of DHEAS on uptake of E(2)17betaG, the attenuation of this effect at high DHEAS concentrations and the lack of reciprocal promotion of DHEAS uptake by E(2)17betaG, a model involving nonreciprocal constructive interactions between some transport substrates is invoked. These results suggest that MRP8 participates in physiological processes involving bile acids, conjugated steroids, and cyclic nucleotides and indicate that the pump has complex interactions with its substrates. PMID- 15537868 TI - Inhibition of L-type Cav1.2 Ca2+ channels by 2,(4-morpholinyl)-8-phenyl-4H-1 benzopyran-4-one (LY294002) and 2-[1-(3-dimethyl-aminopropyl)-5-methoxyindol-3 yl]-3-(1H-indol-3-yl) maleimide (Go6983). AB - Phosphatidylinositol 3-kinase (PI3-K) is involved in physiological processes of cellular proliferation and inflammation and, as postulated recently, in the regulation of L-type Ca(2+) channels. The latter conclusion arose in part from the inhibitory action of the compound 2,(4-morpholinyl)-8-phenyl-4H-1-benzopyran 4-one (LY294002), which has been established as a selective PI3-K inhibitor (IC(50) = 1.4 microM). Herein we show, however, that LY294002 and an inhibitor of protein kinase C (PKC), 2-[1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl]-3-(1H indol-3-yl) maleimide (Go6983), act as direct Ca(2+)-channel inhibitors, with IC(50) values of approximately 20 and 10 microM, respectively. Because both drugs are commonly used at concentrations of approximately 10 microM or higher, the interpretation of such experiments is questionable with respect to a regulatory action of PI3-K or PKC on L-type Ca(2+) channels. PMID- 15537869 TI - Nitric oxide-mediated regulation of connexin43 expression and gap junctional intercellular communication in mesangial cells. AB - This study investigated a potential role of nitric oxide (NO) in the regulation of gap junctional intercellular communication (GJIC). Incubation of mesangial cells (MC) with NO donor S-nitroso-N-acetylpenicillamine (SNAP) enhanced both basal and 8-bromo-cAMP-stimulated GJIC as well as expression of gap junction protein connexin43 (Cx43). This potentiating action of SNAP on Cx43 expression was mimicked by two other NO donors and significantly blocked by soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3,-alpha]quinoxalin-1-1. Guanosine 3',5'-cyclic monophosphate (cGMP) analogue 8-bromo-cGMP exerted an effect similar to NO, whereas another cGMP analogue, 8-pCPT-cGMP, which selectively activates cGMP-dependent kinase without affecting cGMP-inhibited phosphodiesterase (PDE3), had no effect. Moreover, the synergistic action of NO on Cx43 expression was completely prevented by protein kinase A inhibitor H89 but not by cGMP-dependent kinase inhibitor Rp-8-Br-PET-cGMP. These results suggested a possible involvement of NO-cAMP interaction via cGMP-mediated inhibition of PDE3. Indeed, PDE3 inhibitor cilostamide caused potentiation of 8-bromo-cAMP elicited elevations of Cx43 expression that is similar to the effect of SNAP, and an elevation of intracellular cAMP was detected in SNAP-treated cells. With the use of genetically engineered reporter MC that express secreted alkaline phosphatase under the control of the cAMP response element, significant potentiation of cAMP-elicited activation of cAMP response element by SNAP was found. This effect was abrogated in the presence of PDE3 inhibitor cilostamide. Taken together, the results suggest that NO is involved in the control of GJIC and Cx43 expression. This effect of NO is due to activation of protein kinase A via cGMP-dependent inhibition of PDE3 activity. PMID- 15537870 TI - A novel mechanism by which hepatocyte growth factor blocks tubular epithelial to mesenchymal transition. AB - Hepatocyte growth factor (HGF) is a potent antifibrotic cytokine that blocks tubular epithelial to mesenchymal transition (EMT) induced by TGF-beta1. However, the underlying mechanism remains largely unknown. This study investigated the signaling events that lead to HGF blockade of the TGF-beta1-initiated EMT. Incubation of human kidney epithelial cells HKC with HGF only marginally affected the expression of TGF-beta1 and its type I and type II receptors, suggesting that disruption of TGF-beta1 signaling likely plays a critical role in mediating HGF inhibition of TGF-beta1 action. However, HGF neither affected TGF-beta1-induced Smad-2 phosphorylation and its subsequent nuclear translocation nor influenced the expression of inhibitory Smad-6 and -7 in tubular epithelial cells. HGF specifically induced the expression of Smad transcriptional co-repressor SnoN but not Ski and TG-interacting factor at both mRNA and protein levels in HKC cells. SnoN physically interacted with activated Smad-2 by forming transcriptionally inactive complex and overrode the profibrotic action of TGF-beta1. In vivo, HGF did not affect Smad-2 activation and its nuclear accumulation in tubular epithelium, but it restored SnoN protein abundance in the fibrotic kidney in obstructive nephropathy. Hence, HGF blocks EMT by antagonizing TGF-beta1's action via upregulating Smad transcriptional co-repressor SnoN expression. These findings not only identify a novel mode of interaction between the signals activated by HGF receptor tyrosine kinase and TGF-beta receptor serine/threonine kinases but also illustrate the feasibility of confining Smad activity as an effective strategy for blocking renal fibrosis. PMID- 15537871 TI - Decreased signs of nicotine withdrawal in mice null for the beta4 nicotinic acetylcholine receptor subunit. AB - Withdrawal from chronic exposure to nicotine, the main addictive component of tobacco, produces distinctive symptoms in humans. The appearance of these symptoms is a major deterrent when people try to quit smoking. To study which type of nicotine receptor is relevant for the onset of the withdrawal syndrome, we used a mouse model of nicotine withdrawal. Wild-type mice and mice null for the beta4 (beta4-/-) or the beta2 (beta2-/-) nicotinic acetylcholine receptor subunits were implanted with osmotic minipumps delivering 24 mg x kg(-1) x d(-1) nicotine for 13 d. Subsequently, a single intraperitoneal injection of the nicotinic receptor antagonist mecamylamine induced behavioral symptoms of withdrawal measured as increased grooming, chewing, scratching, and shaking, plus the appearance of some unique behaviors such as jumping, leg tremors, and cage scratching. Mecamylamine injection triggered comparable withdrawal signs in wild type and in beta2-/- mice, whereas the beta4-/- mice displayed significantly milder somatic symptoms. In addition, nicotine withdrawal produced hyperalgesia in wild-type but not beta4-/- mice. Finally, chronic nicotine produced an increase in epibatidine binding in several areas of the brain in both wild-type and in beta4-/- mice, but such receptor upregulation did not correlate with the severity of withdrawal signs. Our results demonstrate a major role for beta4 containing nicotinic acetylcholine receptors in the appearance of nicotine withdrawal symptoms. In contrast, the beta2 subunit does not seem to greatly influence this phenomenon. We also show that the upregulation of epibatidine binding sites attributable to chronic nicotine, an effect associated with beta2 containing receptors, is probably not related to the mechanisms underlying withdrawal. PMID- 15537872 TI - Highly efficient small interfering RNA delivery to primary mammalian neurons induces MicroRNA-like effects before mRNA degradation. AB - The study of protein function in neurons has been hindered by the lack of highly efficient, nontoxic methods of inducing RNA interference in such cells. Here we show that application of synthetic small interfering RNA (siRNA) linked to the vector peptide Penetratin1 results in rapid, highly efficient uptake of siRNA by entire populations of cultured primary mammalian hippocampal and sympathetic neurons. This treatment leads to specific knock-down of targeted proteins within hours without the toxicity associated with transfection. In contrast to current methods, our technique permits study of protein function across entire populations with minimal disturbance of complex cellular networks. Using this technique, we found that protein knock-down (evident after 6 hr) precedes any decrease in targeted message (evident after 24 hr), suggesting an early, translational repression by perfectly targeted siRNAs. PMID- 15537873 TI - Independent coding of movement direction and reward prediction by single pallidal neurons. AB - Associating action with its reward value is a basic ability needed by adaptive organisms and requires the convergence of limbic, motor, and associative information. To chart the basal ganglia (BG) involvement in this association, we recorded the activity of 61 well isolated neurons in the external segment of the globus pallidus (GPe) of two monkeys performing a probabilistic visuomotor task. Our results indicate that most (96%) neurons responded to multiple phases of the task. The activity of many (34%) pallidal neurons was modulated solely by direction of movement, and the activity of only a few (3%) pallidal neurons was modulated exclusively by reward prediction. However, the activity of a large number (41%) of single pallidal neurons was comodulated by both expected trial outcome and direction of arm movement. The information carried by the neuronal activity of single pallidal neurons dynamically changed as the trial progressed. The activity was predominantly modulated by both outcome prediction and future movement direction at the beginning of trials and became modulated mainly by movement-direction toward the end of trials. GPe neurons can either increase or decrease their discharge rate in response to predicted future reward. The effects of movement-direction and reward probability on neural activity are linearly summed and thus reflect two independent modulations of pallidal activity. We propose that GPe neurons are uniquely suited for independent processing of a multitude of parameters. This is enabled by the funnel-structure characteristic of the BG architecture, as well as by the anatomical and physiological properties of GPe neurons. PMID- 15537874 TI - Organ of Corti potentials and the motion of the basilar membrane. AB - During sound stimulation, receptor potentials are generated within the sensory hair cells of the cochlea. Prevailing theory states that outer hair cells use the potential-sensitive motor protein prestin to convert receptor potentials into fast alterations of cellular length or stiffness that boost hearing sensitivity almost 1000-fold. However, receptor potentials are attenuated by the filter formed by the capacitance and resistance of the membrane of the cell. This attenuation would limit cellular motility at high stimulus frequencies, rendering the above scheme ineffective. Therefore, Dallos and Evans (1995a) proposed that extracellular potential changes within the organ of Corti could drive cellular motor proteins. These extracellular potentials are not filtered by the membrane. To test this theory, both electric potentials inside the organ of Corti and basilar membrane vibration were measured in response to acoustic stimulation. Vibrations were measured at sites very close to those interrogated by the recording electrode using laser interferometry. Close comparison of the measured electrical and mechanical tuning curves and time waveforms and their phase relationships revealed that those extracellular potentials indeed could drive outer hair cell motors. However, to achieve the sharp frequency tuning that characterizes the basilar membrane, additional mechanical processing must occur inside the organ of Corti. PMID- 15537875 TI - Axonal regeneration and lack of astrocytic gliosis in EphA4-deficient mice. AB - Spinal cord injury usually results in permanent paralysis because of lack of regrowth of damaged neurons. Here we demonstrate that adult mice lacking EphA4 ( /-), a molecule essential for correct guidance of spinal cord axons during development, exhibit axonal regeneration and functional recovery after spinal cord hemisection. Anterograde and retrograde tracing showed that axons from multiple pathways, including corticospinal and rubrospinal tracts, crossed the lesion site. EphA4-/- mice recovered stride length, the ability to walk on and climb a grid, and the ability to grasp with the affected hindpaw within 1-3 months of injury. EphA4 expression was upregulated on astrocytes at the lesion site in wild-type mice, whereas astrocytic gliosis and the glial scar were greatly reduced in lesioned EphA4-/- spinal cords. EphA4-/- astrocytes failed to respond to the inflammatory cytokines, interferon-gamma or leukemia inhibitory factor, in vitro. Neurons grown on wild-type astrocytes extended shorter neurites than on EphA4-/- astrocytes, but longer neurites when the astrocyte EphA4 was blocked by monomeric EphrinA5-Fc. Thus, EphA4 regulates two important features of spinal cord injury, axonal inhibition, and astrocytic gliosis. PMID- 15537877 TI - Functional-anatomic correlates of memory retrieval that suggest nontraditional processing roles for multiple distinct regions within posterior parietal cortex. AB - Current theories of posterior parietal cortex (PPC) function emphasize space based attention and motor intention. Imaging studies of long-term memory have demonstrated PPC activation during successful memory retrieval. Here, we explored the relationship between memory processes and classical notions of PPC function. Study 1 investigated old-new recognition using picture and sound stimuli to test whether PPC memory effects were dependent on visuospatial attention. A region lateral to the intraparietal sulcus [inferior parietal lobule complex (IPLC)] and two regions in the medial PPC [precuneus complex (PCC) and posterior cingulate/retrosplenial cortex (pC/Rsp)] showed strong retrieval success effects for both picture and sound stimuli. Study 2 explored a recognition task with varied response contingencies to investigate whether these retrieval success effects are dependent on motor intentions. In one condition, subjects responded to both old and new items. In two other conditions, subjects responded only to old or only to new items. IPLC, PCC, and pC/Rsp continued to show retrieval success effects with similar magnitudes for all response contingencies, including a condition in which no responses were made to old items. In a third study, IPLC and PCC activity was modulated at retrieval based on levels of processing at study, suggesting sensitivity to memory demands. These studies demonstrate that retrieval success effects in lateral and medial PPC regions are not affected by manipulations predicted by classical theories of PPC function but can be modulated by memory-related manipulations. PPC regions thus have prominent response properties associated with memory, which may arise through interactions with medial temporal cortex. PMID- 15537876 TI - Interferon-gamma induced medulloblastoma in the developing cerebellum. AB - We have generated a mouse model system with a high incidence of medulloblastoma, a malignant neoplasm believed to arise from immature precursors of cerebellar granule neurons. These animals ectopically express interferon-gamma (IFN-gamma) in astrocytes in the CNS in a controlled manner, exploiting the tetracycline controllable system. More than 80% of these mice display severe ataxia and develop cerebellar tumors that express synaptophysin, the mouse atonal homolog MATH1, sonic hedgehog (SHH), and Gli1. IFN-gamma-induced tumorigenesis in these mice is associated with increased expression of SHH, and SHH induction and tumorigenesis are dependent on signal transducer and activator of transcription 1 (STAT1). When IFN-gamma expression is shut down with doxycycline at postnatal day 12 (P12), the clinical symptoms dissipate and the mice do not develop tumors, whereas if transgene expression is shut down at P16, the clinical symptoms and tumors progress to lethality, indicating that IFN-gamma is required for tumor induction but not progression. The tumors that occur in the continued presence of IFN-gamma display extensive necrosis and apoptosis as well as macrophage and lymphocytic infiltration, whereas the tumors that develop in mice in which IFN gamma expression is shut down at P16 do not. Thus, IFN-gamma expression in the perinatal period can induce SHH expression and medulloblastoma in the cerebellum by a STAT1-dependent mechanism, and its continued presence appears to promote a host response to the tumor. PMID- 15537879 TI - The brain-derived neurotrophic factor val66met polymorphism and variation in human cortical morphology. AB - A variation in the BDNF gene (val66met) affects the function of BDNF in neurons, predicts variation in human memory, and is associated with several neurological and psychiatric disorders. Here, we show that, in magnetic resonance imaging scans of a large sample of normal individuals, this polymorphism affects the anatomy of the hippocampus and prefrontal cortex, identifying a genetic mechanism of variation in brain morphology related to learning and memory. PMID- 15537878 TI - Assessing the role of GLUK5 and GLUK6 at hippocampal mossy fiber synapses. AB - It has been suggested recently that presynaptic kainate receptors (KARs) are involved in short-term and long-term synaptic plasticity at hippocampal mossy fiber synapses. Using genetic deletion and pharmacology, we here assess the role of GLU(K5) and GLU(K6) in synaptic plasticity at hippocampal mossy fiber synapses. We found that the kainate-induced facilitation was completely abolished in the GLU(K6)-/- mice, whereas it was unaffected in the GLU(K5)-/-. Consistent with this finding, synaptic facilitation was reduced in the GLU(K6)(-/-) and was normal in the GLU(K5)-/-. In agreement with these results and ruling out any compensatory effects in the genetic deletion models, application of the GLU(K5) specific antagonist LY382884 [(3S,4aR,6S,8aR)-6-(4-carboxyphenyl)methyl 1,2,3,4,4a,5,6,7,8,8a-decahydroisoquinoline-3-carboxylic acid] did not affect short-term and long-term synaptic plasticity at the hippocampal mossy fiber synapses. We therefore conclude that the facilitatory effects of kainate on mossy fiber synaptic transmission are mediated by GLU(K6)-containing KARs. PMID- 15537880 TI - Role of neurokinin 3 receptors in supraoptic vasopressin and oxytocin neurons. AB - Neurokinin 3 receptors (NK3-Rs) are expressed in the supraoptic nucleus (SON), and SON is innervated by substance P (SP)-expressing A1 neurons in the medulla. Because SP stimulates vasopressin (VP) and oxytocin release from explants of the hypothalamo-neurohypophyseal system (HNS), two hypotheses were tested: (1) SP stimulated VP release is mediated by NK3-Rs, and (2) stimulation of the A1 pathway by hypotension activates SON NK3-Rs. Senktide, an NK3-R agonist, stimulated VP release from HNS explants, but neither a neurokinin 1 receptor antagonist [L732,138 (N-acetyl-L-tryptophan 3,5-bis(tri-fluoromethyl)benzyl ester)] nor two NK3-R antagonists (SB222200 and SB235375) prevented SP-stimulated VP release. Because the affinity of these antagonists for rat NK-Rs may limit their efficacy, NK3-R internalization was used to assess the ability of SP to activate SON NK3-Rs. Senktide, SP, or vehicle was microinjected above SON. The brain was perfused 5 min after injection and stained for NK3-R immunoreactivity. Using confocal microscopy, the number of NK3-R-immunoreactive (-IR) endosomes was counted in a 5.6(2) mu region of cytoplasm in SON neurons. Senktide, but not SP or vehicle, significantly increased the number of NK3-R-IR endosomes in the cytoplasm. When hypotension was induced with hydralazine, NK3-R internalization was observed within 5 min (p < 0.005). A decrease in cytoplasmic NK3-R immunoreactivity was observed within 15 min of hypotension. Unexpectedly, both senktide and hypotension resulted in translocation of NK3-R-IR immunoreactivity to the nucleus. Thus, although these studies do not identify SP as the NK3-R ligand, they do provide evidence for hypotension-induced release of an endogenous tachykinin in SON and evidence suggesting a role for NK3-Rs in transcription regulation. PMID- 15537881 TI - Plasticity in the entorhinal cortex suppresses memory for contextual fear. AB - Several studies have delineated a role for the hippocampus in fear conditioning. However, in this task, the role of the entorhinal cortex (EC), the main input output structure for the hippocampus, is uncertain. The extracellular signal regulated kinase (ERK) cascade has been shown to be a molecular correlate for long-term memory, and its activity is required for various types of memory storage, including fear memory. In this study, we show that ERK activity in the EC increased 90 min after fear conditioning. Post-training intra-EC infusion of ERK cascade inhibitors (PD098059, UO126) at 40 min, but not at 10 min, resulted in increased freezing to the context, but not to the tone, during a 48 hr retention test. Interestingly, both PD098059- and UO126-infused animals also demonstrated anticipatory freezing in the context, freezing maximally at the time the shock was given during training. This anticipatory behavior was also seen in naive animals receiving additional training. Together, these results suggest that ERK-mediated plasticity in the EC normally suppresses context-specific fear memory, especially the temporal nature of the freezing response, and that blocking this plasticity mimics the effects of additional training. PMID- 15537882 TI - M2 muscarinic acetylcholine receptor knock-out mice show deficits in behavioral flexibility, working memory, and hippocampal plasticity. AB - Muscarinic acetylcholine receptors are known to play key roles in facilitating cognitive processes. However, the specific roles of the individual muscarinic receptor subtypes (M1-M5) in learning and memory are not well understood at present. In the present study, we used wild-type (M2+/+) and M2 receptor deficient (M2-/-) mice to examine the potential role of M2 receptors in learning and memory and hippocampal synaptic plasticity. M2-/- mice showed significant deficits in behavioral flexibility and working memory in the Barnes circular maze and the T-maze delayed alternation tests, respectively. The behavioral deficits of M2-/- mice were associated with profound changes in neuronal plasticity studied at the Schaffer-CA1 synapse of hippocampal slices. Strikingly, short-term potentiation (STP) was abolished, and long-term potentiation (LTP) was drastically reduced after high-frequency stimulation of M2-/- hippocampi. Treatment of M2-/- hippocampal slices with the GABA(A) receptor antagonist, bicuculline, restored STP and significantly increased LTP. Whole-cell recordings from CA1 pyramidal cells demonstrated a much stronger disinhibition of GABAergic than glutamatergic transmission in M2-/- hippocampi, which was particularly prominent during stimulus trains. Increased strength of GABAergic inhibition is thus a likely mechanism underlying the impaired synaptic plasticity observed with M2-/- hippocampi. Moreover, the persistent enhancement of excitatory synaptic transmission in CA1 pyramidal cells induced by the transient application of a low concentration of a muscarinic agonist (referred to as LTP(m)) was totally abolished in M2-/- mice. Because impaired muscarinic cholinergic neurotransmission is associated with Alzheimer's disease and normal aging processes, these findings should be of considerable therapeutic relevance. PMID- 15537883 TI - Molecular determinants of ligand selectivity in a vertebrate odorant receptor. AB - The identification of the chemical structure of an odorant by the vertebrate olfactory system is thought to occur through the combinatorial activity from multiple receptors, each tuned to recognize different chemical features. What are the molecular determinants underlying the selectivity of individual odorant receptors for their cognate ligands? To address this question, we performed molecular modeling and site-directed mutagenesis on the ligand-binding region of two orthologous amino acid odorant receptors belonging to the "C family" of G protein-coupled receptors in goldfish and zebrafish. We identified the critical ligand-receptor interactions that afford ligand binding as well as selectivity for different amino acids. Moreover, predictions regarding binding pocket structure allowed us to alter, in a predictable manner, the receptor preferences for different ligands. These results reveal how this class of odorant receptor has evolved to accommodate ligands of varying chemical structure and further illuminate the molecular principles underlying ligand recognition and selectivity in this family of chemosensory receptors. PMID- 15537884 TI - Postsynaptic density protein-95 regulates NMDA channel gating and surface expression. AB - NMDA receptors (NMDARs) colocalize with postsynaptic density protein-95 (PSD-95), a multivalent synaptic scaffolding protein and core component of the postsynaptic density, at excitatory synapses. Although much is known about the identity and properties of scaffolding proteins, little is known about their actions on NMDAR function. Here we show that association of PSD-95 with NMDARs modulates channel gating and surface expression. PSD-95 increases the number of functional channels at the cell surface and channel opening rate of NMDARs, with little or no change in conductance, reversal potential, or mean open time. We show further that PSD 95 increases NMDAR surface expression by increasing the rate of channel insertion and decreasing the rate of channel internalization. The PDZ (PSD-95, discs large, zona occludens-1) binding motif at the distal end of the NR2 C-terminal tail is critical to the actions of PSD-95 on NMDAR function and surface expression. Given that activity bi-directionally modifies synaptic levels of PSD-95, our findings suggest a novel mechanism for activity-dependent regulation of NMDARs at central synapses. PMID- 15537885 TI - Glutamate-evoked alterations of glial and neuronal cell morphology in the guinea pig retina. AB - Neuronal activity is accompanied by transmembranous ion fluxes that cause cell volume changes. In whole mounts of the guinea pig retina, application of glutamate resulted in fast swelling of neuronal cell bodies in the ganglion cell layer (GCL) and the inner nuclear layer (INL) (by approximately 40%) and a concomitant decrease of the thickness of glial cell processes in the inner plexiform layer (IPL) (by approximately 40%) that was accompanied by an elongation of the glial cells, by a thickening of the whole retinal tissue, and by a shrinkage of the extracellular space (by approximately 18%). The half maximal effect of glutamate was observed at approximately 250 mum, after approximately 4 min. The swelling was caused predominantly by AMPA-kainate receptor-mediated influx of Na+ into retinal neurons. Similar but transient morphological alterations were induced by high K+ and dopamine, which caused release of endogenous glutamate and subsequent activation of AMPA-kainate receptors. Apparently, retinal glutamatergic transmission is accompanied by neuronal cell swelling that causes compensatory morphological alterations of glial cells. The effect of dopamine was elicitable only during light adaptation but not in the dark, and glutamate and high K+ induced strong ereffects in the dark than in the light. This suggests that not only the endogenous release of dopamine but also the responsiveness of glutamatergic neurons to dopamine is regulated by light-dark adaptation. Similar morphological alterations (neuronal swelling and decreased glial process thickness) were observed in whole mounts isolated immediately after experimental retinal ischemia, suggesting an involvement of AMPA-kainate receptor activation in putative neurotoxic cell swelling in the postischemic retina. PMID- 15537886 TI - Increased GABA release in the central amygdala of ethanol-dependent rats. AB - The central nucleus of amygdala (CeA) is important in regulating alcohol consumption and plays a major role in the anxiogenic response to ethanol withdrawal. We showed previously that acute ethanol augments GABA(A) receptor mediated IPSPs and IPSCs, possibly by a presynaptic mechanism. Here, we have examined the interaction of acute ethanol with the GABAergic system in chronic ethanol-treated (CET) rats using an in vitro CeA slice preparation and in vivo brain microdialysis. We found that in CeA slices from CET rats, the baseline evoked IPSP and IPSC amplitudes were increased, and paired-pulse facilitation ratios were lower than in naive rats, suggesting an increased GABAergic transmission after chronic ethanol treatment. Interestingly, acute ethanol (5-66 mm) significantly enhanced IPSPs and IPSCs equally in CET and naive rats, indicating a lack of tolerance for this effect of acute ethanol. Analysis of miniature IPSC frequency suggests that the increased GABAergic transmission by both acute and chronic ethanol arises from a presynaptic mechanism involving enhanced vesicular release of GABA. These data are supported by microdialysis studies showing that CET rats presented a fourfold increase in baseline GABA dialysate content compared with naive rats. In vivo administration of ethanol (0.1, 0.3, and 1.0 m) produced a dose-dependent increase in GABA release in the CeA dialysate in both CET and naive rats. These combined findings suggest that acute and chronic ethanol increases GABA release in CeA and support previous reports that the behavioral actions of ethanol are mediated, in part, by increased GABAergic transmission in the CeA. PMID- 15537887 TI - Acid-sensing ion channel 2 contributes a major component to acid-evoked excitatory responses in spiral ganglion neurons and plays a role in noise susceptibility of mice. AB - Ion channels in the degenerin-epithelial sodium channel (DEG-ENaC) family perform diverse functions, including mechanosensation. Here we explored the role of the vertebrate DEG-ENaC protein, acid-sensing ion channel 2 (ASIC2), in auditory transduction. Contributions of ASIC2 to hearing were examined by comparing hearing threshold and noise sensitivity of wild-type and ASIC2 null mice. ASIC2 null mice showed no significant hearing loss, indicating that the ASIC2 was not directly involved in the mechanotransduction of the mammalian cochlea. However, we found that (1) ASIC2 was present in the spiral ganglion (SG) neurons in the adult cochlea and that externally applied protons induced amiloride-sensitive sodium currents and action potentials in SG neurons in vitro, (2) proton-induced responses were greatly reduced in SG neurons obtained from ASIC2 null mice, indicating that activations of ASIC2 contributed a major portion of the proton induced excitatory response in SG neurons, and (3) ASIC2 null mice were considerably more resistant to noise-induced temporary, but not permanent, threshold shifts. Together, these data suggest that ASIC2 contributes to suprathreshold functions of the cochlea. The presence of ASIC2 in SG neurons could provide sensors to directly convert local acidosis to excitatory responses, therefore offering a cellular mechanism linking hearing losses caused by many enigmatic causes (e.g., ischemia or inflammation of the inner ear) to excitotoxicity. PMID- 15537888 TI - Induction of brain-region-specific forms of obesity by agouti. AB - Disruption of melanocortin (MC) signaling, such as by ectopic Agouti overexpression, leads to an obesity syndrome with hyperphagia, obesity, and accelerated body weight gain during high-fat diet. To investigate where in the brain disruption of MC signaling results in obesity, long-term Agouti expression was induced after local injections of recombinant adeno-associated viral particles in selected brain nuclei of adult rats. Agouti expression in the paraventricular nucleus, a hypothalamic region with a high density of MC receptors, induced acute onset hyperphagia and rapid weight gain that persisted for at least 6 weeks. In contrast, obesity and hyperphagia developed with a 3 week delay when Agouti was expressed in the dorsal medial hypothalamus. Agouti expression in the lateral hypothalamus (LH) did not affect food intake and body weight during regular diet, despite the presence of MC receptors in this region. However, during exposure to a high-fat diet, animals with Agouti expression in the LH exhibited a marked increase in body weight. Here we show that the LH is important for the protection against diet-induced obesity by controlling caloric intake during consumption of a high-fat diet. Together, this study provides evidence that different aspects of the Agouti-induced obesity syndrome, such as hyperphagia and diet responsiveness, are mediated by distinct brain regions and opens challenging opportunities for further understanding of pathophysiological processes in the development of the obesity syndrome. PMID- 15537889 TI - Plasticity of the avian song control system in response to localized environmental cues in an equatorial songbird. AB - A striking feature of the vertebrate brain is its plasticity. In high-latitude vertebrates, seasonal plasticity of the brain is driven by ubiquitous photoperiod cues and therefore is highly predictable and synchronous across extensive geographic ranges. A pronounced example of seasonal brain plasticity occurs in the nuclei that regulate song behavior in songbirds. These nuclei are larger in breeding than in nonbreeding birds. In the tropics, photoperiod varies little annually, and other environmental cues important for breeding can show considerable local geographic variability. We investigated whether localized patterns of seasonal breeding in tropical birds are associated with brain plasticity. We studied two populations of rufous-collared sparrows (Zonotrichia capensis) that breed, only 25 km apart, on the equator but out of phase with each other. We measured gonadal activity and the size of song nuclei (high vocal center, robust nucleus of the arcopallium, and area X) during each population's breeding and nonbreeding periods. Breeding males had larger song nuclei and greater gonadal activity than did nonbreeding birds. This plasticity was associated with local environmental cues, such that the two populations exhibit asynchronous changes in brain structure. These results demonstrate the sensitivity of the brain and its ability to use a variety of environmental cues to coordinate seasonal plasticity and reproduction. PMID- 15537890 TI - Prestimulus oscillations enhance psychophysical performance in humans. AB - The presence of various ongoing oscillations in the brain is correlated with behavioral states such as restful wakefulness or drowsiness. However, even when subjects aim to maintain a high level of vigilance, ongoing oscillations exhibit large amplitude variability on time scales of hundreds of milliseconds to seconds, suggesting that the functional state of local cortical networks is continuously changing. How this volatility of ongoing oscillations influences the perception of sensory stimuli has remained essentially unknown. We investigated the relationship between prestimulus neuronal oscillations and the subjects' ability to consciously perceive and react to somatosensory stimuli near the threshold of detection. We show that, for prestimulus oscillations at approximately 10, 20, and 40 Hz detected over the sensorimotor cortex, intermediate amplitudes were associated with the highest probability of conscious detection and the shortest reaction times. In contrast, for 10 and 20 Hz prestimulus oscillations detected over the parietal region, the largest amplitudes were associated with the best performance. Our data indicate that the prestimulus oscillatory activity detected over sensorimotor and parietal cortices has a profound effect on the processing of weak stimuli. Furthermore, the results suggest that ongoing oscillations in sensory cortices may optimize the processing of sensory stimuli with the same mechanism as noise sources in intrinsic stochastic resonance. PMID- 15537891 TI - Synaptic targeting by Alzheimer's-related amyloid beta oligomers. AB - The cognitive hallmark of early Alzheimer's disease (AD) is an extraordinary inability to form new memories. For many years, this dementia was attributed to nerve-cell death induced by deposits of fibrillar amyloid beta (Abeta). A newer hypothesis has emerged, however, in which early memory loss is considered a synapse failure caused by soluble Abeta oligomers. Such oligomers rapidly block long-term potentiation, a classic experimental paradigm for synaptic plasticity, and they are strikingly elevated in AD brain tissue and transgenic-mouse AD models. The current work characterizes the manner in which Abeta oligomers attack neurons. Antibodies raised against synthetic oligomers applied to AD brain sections were found to give diffuse stain around neuronal cell bodies, suggestive of a dendritic pattern, whereas soluble brain extracts showed robust AD-dependent reactivity in dot immunoblots. Antigens in unfractionated AD extracts attached with specificity to cultured rat hippocampal neurons, binding within dendritic arbors at discrete puncta. Crude fractionation showed ligand size to be between 10 and 100 kDa. Synthetic Abeta oligomers of the same size gave identical punctate binding, which was highly selective for particular neurons. Image analysis by confocal double-label immunofluorescence established that >90% of the punctate oligomer binding sites colocalized with the synaptic marker PSD-95 (postsynaptic density protein 95). Synaptic binding was accompanied by ectopic induction of Arc, a synaptic immediate-early gene, the overexpression of which has been linked to dysfunctional learning. Results suggest the hypothesis that targeting and functional disruption of particular synapses by Abeta oligomers may provide a molecular basis for the specific loss of memory function in early AD. PMID- 15537892 TI - Insulin-like growth factor-I accelerates the cell cycle by decreasing G1 phase length and increases cell cycle reentry in the embryonic cerebral cortex. AB - Neurogenesis in the developing cerebral cortex of mice occurs in the dorsal telencephalon between embryonic day 11 (E11) and E17, during which time the majority of cortical projection neurons and some glia are produced from proliferating neuroepithelial cells in the ventricular zone. The number of cells produced by this process is governed by several factors, including cell cycle kinetics and the proportion of daughter cells exiting the cell cycle after a given round of cell division. The in vivo effects of IGF-I on cell cycle kinetics were investigated in nestin/IGF-I transgenic (Tg) embryos, in which IGF-I is overexpressed in the cerebral cortex and dorsal telencephalon. These Tg mice have been shown to exhibit increased cell number in the cortical plate by E16 and increased numbers of neurons and glia in the cerebral cortex during postnatal development. Cumulative S phase labeling with 5-bromo-2'-deoxyuridine revealed a decrease in total cell cycle length (TC) in Tg embryos on E14. This decrease in TC was found to result entirely from a reduction in the length of the G1 phase of the cell cycle from 10.66 to 8.81 hr, with no significant changes in the lengths of the S, G2, and M phases. Additionally, the proportion of daughter cells reentering the cell cycle was significantly increased by 15% in Tg embryos on E14 E15 compared with littermate controls. These data demonstrate that IGF-I regulates progenitor cell division in the ventricular zone by reducing G1 phase length and decreasing TC but increases cell cycle reentry. PMID- 15537894 TI - Cortical areas involved in object, background, and object-background processing revealed with functional magnetic resonance adaptation. AB - Previous work has suggested that object and place processing are neuroanatomically dissociated in ventral visual areas under conditions of passive viewing. It has also been shown that the hippocampus and parahippocampal gyrus mediate the integration of objects with background scenes in functional imaging studies, but only when encoding or retrieval processes have been directed toward the relevant stimuli. Using functional magnetic resonance adaptation, we demonstrated that object, background scene, and contextual integration of selectively repeated objects and background scenes could be dissociated during the passive viewing of naturalistic pictures involving object-scene pairings. Specifically, bilateral fusiform areas showed adaptation to object repetition, regardless of whether the associated scene was novel or repeated, suggesting sensitivity to object processing. Bilateral parahippocampal regions showed adaptation to background scene repetition, regardless of whether the focal object was novel or repeated, suggesting selectivity for background scene processing. Finally, bilateral parahippocampal regions distinct from those involved in scene processing and the right hippocampus showed adaptation only when the unique pairing of object with background scene was repeated, suggesting that these regions perform binding operations. PMID- 15537893 TI - Role of mitogen-activated protein kinase activation in injured and intact primary afferent neurons for mechanical and heat hypersensitivity after spinal nerve ligation. AB - To investigate whether activation of mitogen-activated protein kinase (MAPK) in damaged and/or undamaged primary afferents participates in neuropathic pain after partial nerve injury, we examined the phosphorylation of extracellular signal regulated protein kinase (ERK), p38 MAPK, and c-Jun N-terminal kinase (JNK) in the L4 and L5 dorsal root ganglion (DRG) in the L5 spinal nerve ligation (SNL) model. We first confirmed, using activating transcription factor 3 and neuropeptide Y immunoreactivity, that virtually all L4 DRG neurons are spared from axotomy in this model. In the injured L5 DRG, the L5 SNL induced the activation of ERK, p38, and JNK in different populations of DRG neurons. In contrast, in the uninjured L4 DRG, the L5 SNL induced only p38 activation in tyrosine kinase A-expressing small- to medium-diameter neurons. Intrathecal ERK, p38, and JNK inhibitor infusions reversed SNL-induced mechanical allodynia, whereas only p38 inhibitor application attenuated SNL-induced thermal hyperalgesia. Furthermore, the L5 dorsal rhizotomy did not prevent SNL-induced thermal hyperalgesia. We therefore hypothesized that p38 activation in the uninjured L4 DRG might be involved in the development of heat hypersensitivity in the L5 SNL model. In fact, the treatment of the p38 inhibitor and also anti-nerve growth factor reduced SNL-induced upregulation of brain-derived neurotrophic factor and transient receptor potential vanilloid type 1 expression in the L4 DRG. Together, our results demonstrate that the L5 SNL induces differential activation of MAPK in injured and uninjured DRG neurons and, furthermore, that MAPK activation in the primary afferents may participate in generating pain hypersensitivity after partial nerve injury. PMID- 15537895 TI - Improved behavior and neuropathology in the mouse model of Sanfilippo type IIIB disease after adeno-associated virus-mediated gene transfer in the striatum. AB - Sanfilippo syndrome is a mucopolysaccharidosis (MPS) caused by a lysosomal enzyme defect interrupting the degradation pathway of heparan sulfates. Affected children develop hyperactivity, aggressiveness, delayed development, and severe neuropathology. We observed relevant behaviors in the mouse model of Sanfilippo syndrome type B (MPSIIIB), in which the gene coding for alpha-N acetylglucosaminidase (NaGlu) is invalidated. We addressed the feasibility of gene therapy in these animals. Vectors derived from adeno-associated virus serotype 2 (AAV2) or 5 (AAV5) coding for NaGlu were injected at a single site in the putamen of 45 6-week-old MPSIIIB mice. Normal behavior was observed in treated mice. High NaGlu activity, far above physiological levels, was measured in the brain and persisted at 38 weeks of age. NaGlu immunoreactivity was detected in neuron intracellular organelles, including lysosomes. Enzyme activity spread beyond vector diffusion areas. Delivery to the entire brain was reproducibly obtained with both vector types. NaGlu activity was higher and distribution was broader with AAV5-NaGlu than with AAV2-NaGlu vectors. The compensatory increase in the activity of various lysosomal enzymes was improved. The accumulation of gangliosides GM2 and GM3 present before treatment and possibly participating in neuropathology was reversed. Characteristic vacuolations in microglia, perivascular cells, and neurons, which were prominent before the age of treatment, disappeared in areas in which NaGlu was present. However, improvement was only partial in some animals, in contrast to high NaGlu activity. These results indicate that NaGlu delivery from intracerebral sources has the capacity to alleviate most disease manifestations in the MPSIIIB mouse model. PMID- 15537896 TI - Extracellular signal-regulated kinase 1/2 signaling pathway in solitary nucleus mediates cholecystokinin-induced suppression of food intake in rats. AB - Increased food intake is a major factor in the development of obesity, and the control of meal size is a valid approach to reduce food intake in humans. Meal termination, or satiety, is thought to be organized within the caudal brainstem where direct signals from the food handling alimentary canal and long-term signals from the forebrain converge in the solitary nucleus. Cholecystokinin (CCK) released from the gut after ingestion of food has been strongly implicated in nucleus tractus solitarius (NTS)-mediated satiation, but the exact cellular and intracellular signaling events are not understood. Using Western blotting and immunohistochemistry with phosphospecific antibodies, we demonstrate here that peripheral administration of CCK in rats leads to rapid activation of the extracellular signal-regulated kinase (ERK) signaling cascade in NTS neurons and that blockade of ERK signaling with microinfusion of a selective mitogen activated ERK kinase inhibitor into the fourth ventricle attenuates the capacity of CCK to suppress food intake. In addition, we show that CCK-induced activation of ERK results in phosphorylation of the voltage-dependent potassium channel Kv4.2 and the nuclear transcription factor CREB (cAMP response element-binding protein). The results demonstrate that ERK signaling is necessary for exogenous CCK to suppress food intake in deprived rats and suggest that this pathway may also be involved in natural satiation and the period of satiety between meals through coupling of ERK activation to both cytosolic and nuclear effector mechanisms that have the potential to confer acute and long-term changes in neuronal functioning. PMID- 15537897 TI - Regulation of the NMDA receptor complex and trafficking by activity-dependent phosphorylation of the NR2B subunit PDZ ligand. AB - Interactions between NMDA receptors (NMDARs) and the PDZ [postsynaptic density-95 (PSD-95)/Discs large/zona occludens-1] domains of PSD-95/SAP90 (synapse associated protein with a molecular weight of 90 kDa) family proteins play important roles in the synaptic targeting and signaling of NMDARs. However, little is known about the mechanisms that regulate these PDZ domain-mediated interactions. Here we show that casein kinase II (CK2) phosphorylates the serine residue (Ser1480) within the C-terminal PDZ ligand (IESDV) of the NR2B subunit of NMDAR in vitro and in vivo. Phosphorylation of Ser1480 disrupts the interaction of NR2B with the PDZ domains of PSD-95 and SAP102 and decreases surface NR2B expression in neurons. Interestingly, activity of the NMDAR and Ca2+/calmodulin dependent protein kinase II regulates CK2 phosphorylation of Ser1480. Furthermore, CK2 colocalizes with NR1 and PSD-95 at synaptic sites. These results indicate that activity-dependent CK2 phosphorylation of the NR2B PDZ ligand regulates the interaction of NMDAR with PSD-95/SAP90 family proteins as well as surface NMDAR expression and may be a critical mechanism for modulating excitatory synaptic function and plasticity. PMID- 15537898 TI - Bitter taste receptors for saccharin and acesulfame K. AB - Weight-conscious subjects and diabetics use the sulfonyl amide sweeteners saccharin and acesulfame K to reduce their calorie and sugar intake. However, the intrinsic bitter aftertaste, which is caused by unknown mechanisms, limits the use of these sweeteners. Here, we show by functional expression experiments in human embryonic kidney cells that saccharin and acesulfame K activate two members of the human TAS2R family (hTAS2R43 and hTAS2R44) at concentrations known to stimulate bitter taste. These receptors are expressed in tongue taste papillae. Moreover, the sweet inhibitor lactisole did not block the responses of cells transfected with TAS2R43 and TAS2R44, whereas it did block the response of cells expressing the sweet taste receptor heteromer hTAS1R2-hTAS1R3. The two receptors were also activated by nanomolar concentrations of aristolochic acid, a purely bitter-tasting compound. Thus, hTAS2R43 and hTAS2R44 function as cognate bitter taste receptors and do not contribute to the sweet taste of saccharin and acesulfame K. Consistent with the in vitro data, cross-adaptation studies in human subjects also support the existence of common receptors for both sulfonyl amide sweeteners. PMID- 15537899 TI - A mutant ataxin-3 putative-cleavage fragment in brains of Machado-Joseph disease patients and transgenic mice is cytotoxic above a critical concentration. AB - Machado-Joseph disease (MJD) is an inherited neurodegenerative disorder caused by ataxin-3 with a polyglutamine expansion. It is proposed that a toxic cleavage fragment of mutant ataxin-3 alternatively spliced isoform mjd1a triggers neurodegeneration, although this fragment has not yet been detected in the brains of MJD patients or in animal models. We have now generated transgenic mice expressing human mutant (Q71) or normal (Q20) ataxin-3 mjd1a under the control of the mouse prion promoter. Q71 transgenic mice expressing mutant ataxin-3 mjd1a above a critical level developed a phenotype similar to MJD including progressive postural instability, gait and limb ataxia, weight loss, premature death, neuronal intranuclear inclusions, and decreased tyrosine hydroxylase-positive neurons in the substantia nigra (determined by unbiased stereology). Q20 transgenic mice had normal behavior and pathology. Brains from sick Q71 transgenic mice contained an abundant mutant ataxin-3 mjd1a putative-cleavage fragment (Fragment), which was scarce in normal Q71 transgenic mice. Reactivity of the Fragment with a panel of antibodies and comigration with truncations of mutant ataxin-3 revealed that it contained residues C terminal to amino acid 221 to include the polyglutamine expansion. A similar portion of mutant ataxin-3 mjd1a expressed in transfected neuroblastoma cells was toxic above a critical concentration. The Fragment was more abundant in two affected brain regions of MJD patients. Thus, we have developed a murine model for mutant ataxin-3 mjd1a toxicity and identified a putative-cleavage fragment of the disease protein in the brains of these transgenic mice and MJD patients that is cytotoxic above a critical concentration. PMID- 15537900 TI - Progesterone attenuates corticotropin-releasing factor-enhanced but not fear potentiated startle via the activity of its neuroactive metabolite, allopregnanolone. AB - Intact female rats and ovariectomized (OVX) rats with different ovarian steroid replacement regimens were tested for changes in corticotropin-releasing factor (CRF)-enhanced startle (increased acoustic startle amplitude after intracerebroventricular infusion of 1 mug of CRF). OVX rats injected with estradiol (E) followed by progesterone (P) showed a blunted CRF-enhanced startle effect compared with OVX and E-injected rats. CRF-enhanced startle also was reduced significantly in lactating females (high endogenous P levels) compared with cycling rats (low to moderate P levels), as well as in non-E-primed rats when P was administered acutely (4 hr before testing) or chronically (7 d P replacement). The ability of P to attenuate CRF-enhanced startle was probably mediated by its metabolite allopregnanolone [tetrahydroprogesterone (THP)], because THP itself had a similar effect, and chronic administration of medroxyprogesterone, which is not metabolized to THP, did not blunt CRF-enhanced startle but instead slightly increased it. These data suggest that P blunts CRF enhanced startle through a mechanism involving its neuroactive metabolite THP, although a role for the P receptor cannot be completely ruled out. Finally, neither chronic P replacement nor acute THP affected fear-potentiated startle, suggesting that P metabolites have an effect on the bed nucleus of the stria terminalis and anxiety rather than on the amygdala and stimulus-specific fear. PMID- 15537901 TI - Images in clinical medicine. Peripartum dissection of the right coronary artery. PMID- 15537902 TI - Improving patient safety--five years after the IOM report. PMID- 15537903 TI - A great case. PMID- 15537904 TI - Cytokine-based therapies for Crohn's disease--new paradigms. PMID- 15537905 TI - Anti-interleukin-12 antibody for active Crohn's disease. AB - BACKGROUND: Crohn's disease is associated with excess cytokine activity mediated by type 1 helper T (Th1) cells. Interleukin-12 is a key cytokine that initiates Th1-mediated inflammatory responses. METHODS: This double-blind trial evaluated the safety and efficacy of a human monoclonal antibody against interleukin-12 (anti-interleukin-12) in 79 patients with active Crohn's disease. Patients were randomly assigned to receive seven weekly subcutaneous injections of 1 mg or 3 mg of anti-interleukin-12 per kilogram of body weight or placebo, with either a four week interval between the first and second injection (Cohort 1) or no interruption between the two injections (Cohort 2). Safety was the primary end point, and the rates of clinical response (defined by a reduction in the score for the Crohn's Disease Activity Index [CDAI] of at least 100 points) and remission (defined by a CDAI score of 150 or less) were secondary end points. RESULTS: Seven weeks of uninterrupted treatment with 3 mg of anti-interleukin-12 per kilogram resulted in higher response rates than did placebo administration (75 percent vs. 25 percent, P=0.03). At 18 weeks of follow-up, the difference in response rates was no longer significant (69 percent vs. 25 percent, P=0.08). Differences in remission rates between the group given 3 mg of anti-interleukin 12 per kilogram and the placebo group in Cohort 2 were not significant at either the end of treatment or the end of follow-up (38 percent and 0 percent, respectively, at both times; P=0.07). There were no significant differences in response rates among the groups in Cohort 1. The rates of adverse events among patients receiving anti-interleukin-12 were similar to those among patients given placebo, except for a higher rate of local reactions at injection sites in the former group. Decreases in the secretion of interleukin-12, interferon-gamma, and tumor necrosis factor alpha by mononuclear cells of the colonic lamina propria accompanied clinical improvement in patients receiving anti-interleukin-12. CONCLUSIONS: Treatment with a monoclonal antibody against interleukin-12 may induce clinical responses and remissions in patients with active Crohn's disease. This treatment is associated with decreases in Th1-mediated inflammatory cytokines at the site of disease. PMID- 15537906 TI - Mutant mitochondrial elongation factor G1 and combined oxidative phosphorylation deficiency. AB - Although most components of the mitochondrial translation apparatus are encoded by nuclear genes, all known molecular defects associated with impaired mitochondrial translation are due to mutations in mitochondrial DNA. We investigated two siblings with a severe defect in mitochondrial translation, reduced levels of oxidative phosphorylation complexes containing mitochondrial DNA (mtDNA)-encoded subunits, and progressive hepatoencephalopathy. We mapped the defective gene to a region on chromosome 3q containing elongation factor G1 (EFG1), which encodes a mitochondrial translation factor. Sequencing of EFG1 revealed a mutation affecting a conserved residue of the guanosine triphosphate (GTP)-binding domain. These results define a new class of gene defects underlying disorders of oxidative phosphorylation. PMID- 15537907 TI - Intermediate filament proteins and their associated diseases. PMID- 15537908 TI - Images in clinical medicine. Staghorn renal-cell carcinoma. PMID- 15537909 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 34-2004. A 45-year-old man with recurrent pain in the right flank and hematuria. PMID- 15537910 TI - Extremely preterm birth and parental authority to refuse treatment--the case of Sidney Miller. PMID- 15537911 TI - Cachexia in cancer--zeroing in on myosin. PMID- 15537912 TI - Primary care physicians who treat blacks and whites. PMID- 15537913 TI - High versus low PEEP in ARDS. PMID- 15537914 TI - Venom immunotherapy. PMID- 15537915 TI - Aldosterone revisited. PMID- 15537916 TI - Deep-vein thrombosis. PMID- 15537917 TI - Oligospermia in a patient receiving imatinib therapy for the hypereosinophilic syndrome. PMID- 15537918 TI - Phosphorus runoff from agricultural land and direct fertilizer effects: a review. AB - Phosphorus (P) is one of the most important mineral nutrients in agricultural systems, and along with nitrogen (N), is generally the most limiting nutrient for plant production. Farming systems have intensified greatly over time, and in recent years it has become apparent that the concomitant increase in losses of N and P from agricultural land is having a serious detrimental effect on water quality and the environment. The last two decades have seen a marked increase in research into the issues surrounding diffuse losses of P to surface and ground water. This paper reviews this research, examining the issue of P forms in runoff, and highlighting the exceptions to some generally held assumptions about land use and P transport. In particular the review focuses on P losses associated with recent P fertilizer application, as opposed to organic manures, both on the amounts and the forms of P in runoff water. The effects of the physicochemical characteristics of different forms of P fertilizer are explored, particularly in relation to water solubility. Various means of mitigating the risk of loss of P are discussed. It is argued that the influence of recent fertilizer applications is an under-researched area, yet may offer the most readily applicable opportunity to mitigate P losses by land users. This review highlights and discusses some options that have recently become available that may make a significant contribution to the task of sustainable management of nutrient losses from agriculture. PMID- 15537919 TI - Phosphorus transfer in surface runoff from intensive pasture systems at various scales: a review. AB - Phosphorus transfer in runoff from intensive pasture systems has been extensively researched at a range of scales. However, integration of data from the range of scales has been limited. This paper presents a conceptual model of P transfer that incorporates landscape effects and reviews the research relating to P transfer at a range of scales in light of this model. The contribution of inorganic P sources to P transfer is relatively well understood, but the contribution of organic P to P transfer is still relatively poorly defined. Phosphorus transfer has been studied at laboratory, profile, plot, field, and watershed scales. The majority of research investigating the processes of P transfer (as distinct from merely quantifying P transfer) has been undertaken at the plot scale. However, there is a growing need to integrate data gathered at a range of scales so that more effective strategies to reduce P transfer can be identified. This has been hindered by the lack of a clear conceptual framework to describe differences in the processes of P transfer at the various scales. The interaction of hydrological (transport) factors with P source factors, and their relationship to scale, require further examination. Runoff-generating areas are highly variable, both temporally and spatially. Improvement in the understanding and identification of these areas will contribute to increased effectiveness of strategies aimed at reducing P transfers in runoff. A thorough consideration of scale effects using the conceptual model of P transfer outlined in this paper will facilitate the development of improved strategies for reducing P losses in runoff. PMID- 15537920 TI - Septic systems as potential pollution sources in the Cannonsville Reservoir Watershed, New York. AB - On-site septic systems require appropriate soil characteristics to provide effective wastewater treatment. The objective of this study was to evaluate siting practices and treatment efficacy of on-site septic systems within the Cannonsville Reservoir watershed (115900 ha) in the state of New York. Using digital soil survey data, a database of on-site conditions was developed from more than 1100 existing septic system siting records. Soil map units were grouped into four classes based on their suitability to meet common septic system design criteria. A geographic information system was found to be a useful tool for assessment and visual display of septic system and landscape information. Geographic information system analysis indicated that while 80% of soils in the watershed were found to have characteristics that interfere with septic system function, 69% of septic systems installed were of designs suited for soils with no or few restrictive parameters. Since the designs of many septic systems have relied heavily on horizontal distance to streams (mean = 130 m) to provide adequate treatment, potential failures would lead to discharge of compounds of environmental concern, such as phosphorus, with public health implications. The results imply that many septic systems functioning in the watershed are in need of design improvements. PMID- 15537921 TI - An economic evaluation of livestock odor regulation distances. AB - Setback regulations-legislated distances that livestock production facilities must be removed from surrounding properties-are meant to mitigate odor impacts. If the setback length is too short, then there is evidence that surrounding properties and people suffer uncompensated damages. If, on the other hand, setback lengths are too long, then livestock producers may be paying more than that required to compensate for odor-related environmental damages. The purpose of this study is to assess the impact of Kentucky's livestock production facility setbacks on the value of surrounding properties and farm financial management decisions. This paper develops a model of the benefits of livestock odor reduction and the livestock odor abatement cost associated with setback lengths paid by producers. The results of this investigation indicate that the mandated setback lengths for Kentucky are too short. Livestock production firms are worse off under longer setback lengths, but the losses to surrounding home owners far exceed the firm gains at the mandated setbacks. A finding of this study is that the firm has no incentive to completely protect the legislated setback length. Livestock producers in compliance with the relevant setback length may feel protected from odor lawsuits despite damage being done to surrounding property. This suggests that the perceived threat of lawsuit is currently low in the state of Kentucky. Both industry and public goals could be met from further research including location and economic impact of livestock production. PMID- 15537922 TI - Monitoring nitrogen deposition in throughfall using ion exchange resin columns: a field test in the San Bernardino Mountains. AB - Conventional throughfall collection methods are labor intensive and analytically expensive to implement at broad scales. This study was conducted to test an alternative approach requiring infrequent sample collection and a greatly reduced number of chemical analyses. The major objective of the study was to determine the feasibility of using ion exchange resin (IER) to measure N deposition in throughfall with field deployment periods of 3 to 12 mo. Nitrogen deposition measurements in bulk throughfall collected under pine (Pinus sp.) canopies and in forest clearings were compared between co-located conventional throughfall solution collectors and IER throughfall collectors using mixed bed IER columns. Deposition data were collected for 1 yr at a high deposition site (Camp Paivika, CP) and a relatively low one (Barton Flats, BF) in the San Bernardino Mountains in southern California: Annual throughfall deposition values (kg ha(-1) of NH(4) N + NO(3)-N) under large ponderosa pine trees (Pinus ponderosa Laws.) were 145.8 and 143.9 at CP and 17.0 and 15.0 at BF according to the IER and conventional methods, respectively. Analogous values for bulk deposition in forest clearings were 15.6 and 12.3 at CP and 4.0 and 3.3 at BF. It was concluded that the IER collectors can be used for routine monitoring of deposition in throughfall and bulk deposition, provided that field blanks are used to account for background levels of N in the IER columns, which at times are slightly elevated, possibly from slow release of amine groups from the anion exchange resin during field exposures. PMID- 15537923 TI - Effect of surfactants on solubilization and degradation of phenanthrene under thermophilic conditions. AB - The bioavailability and biodegradation of polycyclic aromatic hydrocarbons (PAHs) can be increased through the addition of surfactants. Previous studies of this nature have been conducted under mesophilic conditions. Hence, the aim of the present study was to investigate the effects of synthetic surfactants and biosurfactants on solubilization and degradation of phenanthrene (PHE) in a series of batch solution experiments under thermophilic conditions. Tween 80, Triton X-100, and biosurfactants produced from Pseudomonas aeruginosa strain P CG3 (P-CG3) and Pseudomonas aeruginosa ATCC 9027 (P. 9027) were used in this study. Surfactants effectively enhanced the solubility of PHE at 50 degrees C and the biosurfactant from P-CG3 was most effective with a 28-fold increase in apparent solubility of PHE at a concentration of 10 x critical micelle concentration (CMC) compared with the controls. However, addition of synthetic surfactants or biosurfactants inhibited the biodegradation of PHE in mineral salts medium by an isolate Bacillus sp. B-UM. Degradation of PHE diminished with increasing surfactant concentrations, and PHE degradation was completely inhibited for all the surfactants tested when the concentrations were greater than their respective CMC. The growth test suggested that Tween 80 and biosurfactants were degradable, but preferential utilization of these surfactants as substrates was not the mechanism for explaining the inhibition of PHE biodegradation. Because of the hydrophobic property of B-UM, degradation inhibition of PHE by surfactants was probably due to the reduction of direct contact between bacterial cells and PHE. PMID- 15537924 TI - Biological reduction of chlorate in a gas-lift reactor using hydrogen as an energy source. AB - Chlorate release into the environment occurs with its manufacture and use. Biological reduction of chlorate offers an attractive option to decrease this release. A hydrogen gas-lift reactor with microorganisms attached to pumice particles was used for the treatment of wastewater containing high concentrations of chlorate. The microorganisms used chlorate as an electron acceptor and hydrogen gas as a reducing agent. After a start-up period of only a few weeks, chlorate reduction rates of 3.2 mmol L(-1) h(-1) were achieved during continuous operation. During this period, a hydrogen consumption rate of 14.5 mmol L(-1) h( 1) was observed. Complete removal of chlorate was maintained at hydraulic retention times of 6 h. This study clearly demonstrates the potential of hydrogen gas-lift bioreactors for the treatment of chlorate-containing waste streams. PMID- 15537925 TI - Variation in root density along stream banks. AB - While it is recognized that vegetation plays a significant role in stream bank stabilization, the effects are not fully quantified. The study goal was to determine the type and density of vegetation that provides the greatest protection against stream bank erosion by determining the density of roots in stream banks. To quantify the density of roots along alluvial stream banks, 25 field sites in the Appalachian Mountains were sampled. The riparian buffers varied from short turfgrass to mature riparian forests, representing a range of vegetation types. Root length density (RLD) with depth and aboveground vegetation density were measured. The sites were divided into forested and herbaceous groups and differences in root density were evaluated. At the herbaceous sites, very fine roots (diameter < 0.5 mm) were most common and more than 75% of all roots were concentrated in the upper 30 cm of the stream bank. Under forested vegetation, fine roots (0.5 mm < diameter < 2.0 mm) were more common throughout the bank profile, with 55% of all roots in the top 30 cm. In the top 30 cm of the bank, herbaceous sites had significantly greater overall RLD than forested sites (alpha = 0.01). While there were no significant differences in total RLD below 30 cm, forested sites had significantly greater concentrations of fine roots, as compared with herbaceous sites (alpha = 0.01). As research has shown that erosion resistance has a direct relationship with fine root density, forested vegetation may provide better protection against stream bank erosion. PMID- 15537926 TI - The reduction of internal phosphorus loading using alum in Spring Lake, Michigan. AB - The release of P from lake sediments, which occurs as a part of internal loading, may contribute a significant portion of the total P load to a lake. Phosphorus release rates from sediments in Spring Lake, Michigan, and the degree to which alum reduces P release from these sediments, were investigated during the summer of 2003. Triplicate sediment cores were sampled from four sites in the lake, and exposed to one of four treatments in the laboratory: (i) aerobic water column/alum, (ii) aerobic water column/no alum, (iii) anaerobic water column/alum, or (iv) anaerobic water column/no alum. Total P (TP) release rates were virtually undetectable in the alum treatments (both aerobic and anaerobic). Low, but detectable, release rates were measured in the aerobic/no alum treatment. The highest release rates were measured in the anaerobic/no alum treatments, and ranged from 1.6 to 29.5 mg P m(-2) d(-1) depending on how the calculations were derived. These fluxes translated to mean internal loads that ranged between 2.7 (low range) and 6.4 (high range) Mg yr(-1) when extrapolated to a whole-lake basis. Internal P loads accounted for between 55 and 65% of the total P load to Spring Lake. Although alum is a potentially effective means of reducing the sediment source of P, there is considerable uncertainty in how long an alum treatment would remain effective in this system given the current rates of external loading and the lack of information on wind-wave action and bioturbation in Spring Lake. PMID- 15537927 TI - Temporal changes in soil partitioning and bioaccessibility of arsenic, chromium, and lead. AB - The hazard imposed by trace element contaminants within soils is dependent on their ability to migrate into water systems and their availability for biological uptake. The degree to which a contaminant may dissociate from soil solids and become available to a target organism (i.e., bioaccessibility) is therefore a determining risk factor. We used a physiologically based extraction test (PBET) to estimate the bioaccessible fraction of arsenic-, chromium-, and lead-amended soil. We investigated soils from the A and B horizons of the Melton Valley series, obtained from Oak Ridge National Laboratory site, to address temporal changes in bioaccessibility. Additionally, common extractions that seek to define reactive pools of metals were employed and their correlation to PBET levels evaluated. With the exception of Pb amended to the A horizon, all other treatments exhibited an exponential decrease in bioaccessibility with incubation time. The bioaccessible fraction was less than 0.2 mg kg(-1) within 30 d of incubation for As and Cr in the A horizon and for As and Pb within the B horizon; Cr in the B horizon declined to nearly 0.3 mg kg(-1) within 100 d of aging. The exchangeable fraction declined with incubation period and, with the exception of Pb, was highly correlated with the decline in bioaccessibility. Our results demonstrate limited bioaccessibility in all but one case and the need to address both short-term temporal changes and, most importantly, the soil physiochemical properties. They further reveal the importance of incubation time on the reactivity of such trace elements. PMID- 15537928 TI - Near- and mid-infrared diffuse reflectance spectroscopy for measuring soil metal content. AB - Rapid and nondestructive methods such as diffuse reflectance infrared spectroscopy provide potentially useful alternatives to time-consuming chemical methods of soil metal analysis. To assess the utility of near-infrared reflectance spectroscopy (NIRS) and diffuse mid-infrared reflectance spectroscopy (DRIFTS) for soil metal determination, 70 soil samples from the metal mining region of Tarnowskie Gory (Upper Silesia, Poland) were analyzed by both chemical and spectroscopic methods. Soils represented a wide range of pH (4.0-8.0), total carbon (5.1-73.2 g kg(-1)), and textural classes (from sand to silty clay loam). Soils had various contents of metals (14-4500 mg kg(-1) for Zn, 18-6530 mg kg(-1) for Pb, and 0.17-34 mg kg(-1) for Cd), ranging from natural background levels to high contents indicative of industrial contamination in the region. Soil samples were scanned at the wavelengths from 400 to 2498 nm (near-infrared region) and from 2500 to 25000 nm (mid-infrared region). Calibrations were developed using the one-out validation procedure under partial least squares (PLS) regression. Mid-infrared spectroscopy markedly outperformed NIRS. Iron, Cd, Cu, Ni, and Zn were successfully predicted using DRIFTS. The coefficients of determination (R(2)) between actual and predicted contents were 0.97, 0.94, 0.80, 0.99, and 0.96 for those metals, respectively. Only Pb content was predicted poorly. Calibrations using NIRS were less accurate. Root mean squared deviation (RMSD) values were from 1.27 (Pb) to 3.3 (Ni) times higher for NIRS than for DRIFTS. Results indicate that DRIFTS may be useful for accurate predictions of metals if samples originate from one region. PMID- 15537929 TI - Mechanisms of thorium migration in a semiarid soil. AB - Thorium concentrations at Kirtland Air Force Base training sites in Albuquerque, NM, have been previously described; however, the mechanisms of thorium migration were not fully understood. This work describes the processes affecting thorium mobility in this semiarid soil, which has implications for future remedial action. Aqueous extraction and filtration experiments have demonstrated the colloidal nature of thorium in the soil, due in part to the low solubility of thorium oxide. Colloidal material was defined as that removed by a 0.22-microm or smaller filter after being filtered to nominally dissolved size (0.45 microm). Additionally, association of thorium with natural organic matter is suggested by micro- and ultrafiltration methods, and electrokinetic data, which indicate thorium migration as a negatively charged particle or anionic complex with organic matter. Soil fractionation and digestion experiments show a bimodal distribution of thorium in the largest and smallest size fractions, most likely associated with detrital plant material and inorganic oxide particles, respectively. Plant uptake studies suggest this could also be a mode of thorium migration as plants grown in thorium-containing soil had a higher thorium concentration than those in control soils. Soil erosion laboratory experiments with wind and surface water overflow were performed to determine bulk soil material movement as a possible mechanism of mobility. Information from these experiments is being used to determine viable soil stabilization techniques at the site to maintain a usable training facility with minimal environmental impact. PMID- 15537930 TI - Trace element concentrations in soil, corn leaves, and grain after cessation of biosolids applications. AB - From 1974 to 1984, 543 Mg ha(-1) of biosolids were applied to portions of a land reclamation site in Fulton County, IL. Soil organic C increased to 5.1% then decreased significantly (p < 0.01) to 3.8% following cessation of biosolids applications (1985-1997). Metal concentrations in amended soils (1995-1997) were not significantly different (p > 0.05) (Ni and Zn) or were significantly lower (p < 0.05) (6.4% for Cd and 8.4% for Cu) than concentrations from 1985-1987. For the same biosolids-amended fields, metal concentrations in corn (Zea mays L.) either remained the same (p > 0.05, grain Cu and Zn) or decreased (p < 0.05, grain Cd and Ni, leaf Cd, Cu, Ni, Zn) for plants grown in 1995-1997 compared with plants grown immediately following termination of biosolids applications (1985-1987). Biosolids application increased (p < 0.05) Cd and Zn concentrations in grain compared with unamended fields (0.01 to 0.10 mg kg(-1) for Cd and 23 to 28 mg kg( 1) for Zn) but had no effect (p > 0.05) on grain Ni concentrations. Biosolids reduced (p < 0.05) Cu concentration in grain compared with grain from unamended fields (1.9 to 1.5 mg kg(-1)). Biosolids increased (p < 0.05) Cd, Ni, and Zn concentrations in leaves compared with unamended fields (0.3 to 5.6 mg kg(-1) for Cd, 0.2 to 0.5 mg kg(-1) for Ni, and 32 to 87 mg kg(-1) for Zn), but had no significant effect (p > 0.05) on leaf Cu concentrations. Based on results from this field study, USEPA's Part 503 risk model overpredicted transfer of these metals from biosolids-amended soil to corn. PMID- 15537931 TI - The effect of pH on metal accumulation in two Alyssum species. AB - Nickel phytoextraction using hyperaccumulator plants offers a potential for profit while decontaminating soils. Although soil pH is considered a key factor in metal uptake by crops, little is known about soil pH effects on metal uptake by hyperaccumulator plants. Two Ni and Co hyperaccumulators, Alyssum murale and A. corsicum, were grown in Quarry muck (Terric Haplohemist) and Welland (Typic Epiaquoll) soils contaminated by a Ni refinery in Port Colborne, Ontario, Canada, and in the serpentine Brockman soil (Typic Xerochrepts) from Oregon, USA. Soils were acidified and limed to cover pH from strongly acidic to mildly alkaline. Alyssum grown in both industrially contaminated soils exhibited increased Ni concentration in shoots as soil pH increased despite a decrease in water-soluble soil Ni, opposite to that seen with agricultural crop plants. A small decrease in Alyssum shoot Ni concentration as soil pH increased was observed in the serpentine soil. The highest fraction of total soil Ni was phytoextracted from Quarry muck (6.3%), followed by Welland (4.7%), and Brockman (0.84%). Maximum Ni phytoextraction was achieved at pH 7.3, 7.7, and 6.4 in the Quarry, Welland, and Brockman soils, respectively. Cobalt concentrations in shoots increased with soil pH increase in the Quarry muck, but decreased in the Welland soil. Plants extracted 1.71, 0.83, and 0.05% of the total soil Co from Welland, Quarry, and Brockman, respectively. The differences in uptake pattern of Ni and Co by Alyssum from different soils and pH were probably related to the differences in organic matter and iron contents of the soils. PMID- 15537932 TI - Effect of the Prestige oil spill on salt marsh soils on the coast of Galicia (northwestern Spain). AB - At four estuarine sites on the coast of Galicia (northwestern Spain), all of which were affected by the Prestige oil spill, soil samples were taken from polluted and unpolluted areas and their petroleum hydrocarbon contents, heavy metal contents, and other chemical and physical characteristics were measured. Oil pollution altered both chemical and physical soil properties, aggregating soil particles in plaques, lowering porosity, and increasing resistance to penetration and hydrophobicity. The chromium, nickel, copper, iron, lead, and vanadium contents of polluted soils were between 2 and 2500 times higher than those of their unpolluted counterparts and the background concentrations in Galician coastal sediments. In the cases of Cr, Cu, Ni, Pb, and V, their origin in the polluting oil was corroborated by the high correlation (r >/= 0.74) between the concentrations of these metals and the total petroleum hydrocarbon (TPH) content of the polluted soils. Soil redox potentials ranged from -19 to 114 mV in polluted soils and 112 to 164 mV in unpolluted soils, and were negatively correlated with TPH content (p < 0.01). The low values in the polluted soils explain why the soluble fractions of their total heavy metal contents were very small (generally less than 3%, and in many cases undetectable). PMID- 15537933 TI - Particulate phosphorus transport within stream flow of an agricultural catchment. AB - There is interest in quantifying phosphorus (P) loss from intensively grazed dairy landscapes to identify key pathways and target remediation methods. The Bog Burn drains a dairying catchment in Southland, New Zealand, and has been monitored at fortnightly intervals over a 12-mo period at four sites for suspended sediment (SS), dissolved reactive phosphorus (DRP), and total phosphorus (TP). Time-integrated samplers, deployed at 0.6 median water depth at each site (calculated from previous year's flow data), collected sediment samples, which were analyzed for SS, bioavailable phosphorus (BAP), and TP. Mean concentrations of DRP and TP in stream flow and BAP and TP in sediment were generally highest in summer or autumn (0.043 mg DRP L(-1), 0.160 mg TP L(-1), 173 mg BAP kg(-1), 2228 mg TP kg(-1)) and lowest in winter or spring (0.012 mg DRP L( 1), 0.034 mg TP L(-1), 6 mg BAP kg(-1), 711 mg TP kg(-1)), while loads were highest in winter. Analysis of (137)Cs concentrations in trapped sediment, topsoil, subsoil, and stream bed and bank sediment indicated that trapped sediment was derived from topsoil and entered the stream either through tile drainage or, to a lesser extent, overland flow. Because concentrations of DRP and TP in stream flow are in excess of recommended limits for good water quality (>0.01 mg DRP L(-1), 0.033 mg TP L(-1)), management should focus on the topsoil and specifically on decreasing P loss via tile drainage. This is best achieved by decreasing soil Olsen P concentrations, especially because, on average, Olsen P concentrations in the catchment were above the agronomic optimum. PMID- 15537934 TI - Fluometuron and pendimethalin runoff from strip and conventionally tilled cotton in the southern atlantic coastal plain. AB - In the Atlantic Coastal Plain region of southern Georgia (USA), cotton (Gossypium hirsutum L.) acreage increased threefold in the past decade. To more effectively protect water quality in the region, best management practices are needed that reduce pesticide runoff from fields in cotton production. This study compared runoff of two herbicides, fluometuron [N,N-dimethyl-N'-[3-(trifluoromethyl) phenyl]-urea] and pendimethalin [N-(1-ethylpropyl)-3,4-dimethyl-2,6-dinitro benzenamine], from plots in strip-tillage (ST) and conventional-tillage (CT) management near Tifton, GA. Rainfall simulations were conducted one day after preemergence herbicide applications to 0.0006-ha plots and runoff from 0.15-ha plots due to natural rainfall following preemergence pendimethalin and fluometuron and postemergence fluometuron use was monitored. Pendimethalin runoff was greater under CT than ST due to strong pendimethalin soil sorption and higher erosion and runoff under CT. The highest losses, 1.3% of applied in CT and 0.22% of applied in ST, were observed during rainfall simulations conducted 1 DAT. Fluometuron runoff from natural rainfall was substantially lower from ST than from CT plots but the trend was reversed in rainfall simulations. In all studies, fluometuron runoff was also relatively low (<1% of applied), and on plots under natural rainfall, desmethylfluometuron (DMF) represented about 50% of total fluometuron runoff. Fluometuron's relatively low runoff rate appeared linked to its rapid leaching, and high DMF detection rates in runoff support DMF inclusion in fluometuron risk assessments. Results showed that ST has the potential to reduce runoff of both herbicides, but fluometuron leaching may be a ground water quality concern. PMID- 15537935 TI - Refined tunable methodology for characterization of contaminant-particle relationships in surface water. AB - To understand contaminant transport in aquatic systems, it is essential to define the physical characteristics of the primary particulate carriers. The distribution of organic pollutants with particle-size class and particle morphology in a freshwater embayment (Hamilton Harbor, western Lake Ontario) was studied using a sequence of novel sample preparation and characterization techniques. Water samples (24 L) were fractionated according to particle-size distribution using differential cascade sedimentation and centrifugation methods. These size fractions were subsequently subjected to a physicochemical characterization using scanning transmission electron microscopy and energy dispersive spectroscopy to identify flocs and individual colloidal particles in the size range of 1 nm to 1 mm in diameter. Analytical chemical analyses were performed to identify organic contaminants in extracts prepared from particle size classes, including polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs). The contaminant distribution trends were very similar for all compound classes studied; contaminants were primarily associated with fractions containing particles less than 2 mum in diameter. Morphological characterization of these fractions showed the majority of the particulates to be humic fractals. The results of this study show that contaminants in aquatic systems can be preferentially associated with specific types of particle carriers, the characteristics of which can be clearly defined in terms of size and morphology. PMID- 15537936 TI - Residual and contact herbicide transport through field lysimeters via preferential flow. AB - Usage of glyphosate [N-(phosphonomethyl)-glycine] and glufosinate [2-amino-4 (hydroxy-methylphosphinyl)butanoic acid] may reduce the environmental impact of agriculture because they are more strongly sorbed to soil and may be less toxic than many of the residual herbicides they replace. Preferential flow complicates the picture, because due to this process, even strongly sorbed chemicals can move quickly to ground water. Therefore, four monolith lysimeters (8.1 m(2) by 2.4 m deep) were used to investigate leaching of contact and residual herbicides under a worst-case scenario. Glufosinate, atrazine (6-chloro-N(2)-ethyl-N(4)-isopropyl 1,3,5-triazine-2,4-diamine), alachlor [2-chloro-N-(2,6-diethylphenyl)-N (methoxymethyl) acetamide], and linuron (3-3,4-dichlorophenyl-1-methoxy-1 methylurea) were applied in 1999 before corn (Zea mays L.) planting and glyphosate, alachlor, and metribuzin [4-amino-6-(1,1-dimethylethyl)-3 (methylthio)-1,2,4-triazin-5(4H)-one] were applied in 2000 before soybean [Glycine max (L.) Merr.] planting. A high-intensity rainfall was applied shortly after herbicide application both years. Most alachlor, metribuzin, atrazine, and linuron losses occurred within 1.1 d of rainfall initiation and the peak concentration of the herbicides coincided (within 0.1 d of rainfall initiation in 2000). More of the applied metribuzin leached compared with alachlor during the first 1.1 d after rainfall initiation (2.2% vs. 0.035%, P < 0.05). In 1999, 10 of 24 discrete samples contained atrazine above the maximum contaminant level (atrazine maximum contaminant level [MCL] = 3 mug L(-1)) while only one discrete sample contained glufosinate (19 mug L(-1), estimated MCL = 150 mug L(-1)). The results indicate that because of preferential flow, the breakthrough time of herbicides was independent of their sorptive properties but the transport amount was dependent on the herbicide properties. Even with preferential flow, glyphosate and glufosinate were not transported to 2.4 m at concentrations approaching environmental concern. PMID- 15537937 TI - Distribution and leaching of methyl iodide in soil following emulated shank and drip application. AB - Methyl iodide (MeI) is a promising alternative to methyl bromide in soil fumigation. The pest-control efficacy and ground water contamination risks of MeI as a fumigant are highly related to its gas-phase distribution and leaching after soil application. In this study, the distribution and leaching of MeI in soil following shank injection and subsurface drip application were investigated. Methyl iodide (200 kg ha(-1)) was directly injected or drip-applied at a 20-cm depth into Arlington sandy loam (coarse-loamy, mixed, thermic Haplic Durixeralfs) columns (12-cm i.d., 70-cm height) tarped with virtually impermeable film. Concentration profiles of MeI in the soil air were monitored for 7 d. Methyl iodide diffused rapidly after soil application, and reached a 70-cm depth within 2 h. Relative to shank injection, drip application inhibited diffusion, resulting in significantly lower concentration profiles in the soil air. Seven days after MeI application, fumigated soil was uncapped, aerated for 7 d, and leached with water. Leaching of MeI was significant from the soil columns under both application methods, with concentrations of >10 mug L(-1) in the early leachate. The leaching was greater following shank injection than drip application, with an overall potential of 33 g ha(-1) for shank injection and 19 g ha(-1) for drip application. Persistent residues of MeI remaining in soils after leaching were 50 to 240 ng kg(-1), and the contents were slightly higher following shank injection than drip application. The results suggest that fumigation with MeI may pose a risk of ground water contamination in vulnerable areas. PMID- 15537938 TI - Effect of combined application of methyl isothiocyanate and chloropicrin on their transformation. AB - Combining several soil fumigants to increase the broad spectrum of pest control is a common fumigation practice in current production agriculture. In this study, we investigated the effect of combined application of chloropicrin and methyl isothiocyanate (MITC) on their transformations and persistence in the environment. In aqueous solution, no direct reaction between MITC and chloropicrin occurred and relatively slow rates of hydrolysis of these compounds were observed in aquatic environments free of suspended solids. The transformation of chloropicrin, however, was accelerated in aqueous solution with MITC because of a reduction reaction with bisulfide (HS(-)), which is a by product of MITC hydrolysis. In soil, when fumigants were applied simultaneously, the degradation of MITC was suppressed under the bi-fumigant application due to the inhibition of soil microbial activity and a possible abiotic competition with chloropicrin for a limited number of reaction sites on the surface of soil particles. However, the degradation rate of chloropicrin was significantly enhanced in the bi-fumigant soil system, which was primarily attributed to the reaction of chloropicrin and HS(-). Two sequential application approaches were developed to investigate the feasibility of the combined application of metam sodium (parent compound of MITC) and chloropicrin in soil and assess their potential effects on environmental fate. For both application sequences, the degradation of chloropicrin was accelerated and that of MITC, as a major breakdown product of metam sodium, was inhibited in soil. PMID- 15537939 TI - Remediating RDX-contaminated ground water with permanganate: laboratory investigations for the Pantex perched aquifer. AB - Ground water beneath the U.S. Department of Energy Pantex Plant is contaminated with the high explosive RDX (hexahydro-1,3,5-trinitro-1,3,5 triazine). The USDOE Innovative Treatment and Remediation Demonstration (ITRD) program identified in situ oxidation by permanganate as a technology fit for further investigation. We evaluated the efficacy of KMnO(4) to transform and mineralize RDX by determining degradation kinetics and carbon mass balances using (14)C-RDX. Aqueous RDX solutions (2-5 mg L(-1)) and RDX-contaminated slurries (50% solids, w/v) were treated with KMnO(4) at 1000, 2000, 4000, and 20000 mg L(-1). Treating an aqueous RDX solution of 2.8 mg L(-1) with 20000 mg KMnO(4) L(-1) decreased RDX to 0.1 mg L(-1) within 11 d while cumulative mineralization proceeded for 14 d until 87% of the labeled carbon was trapped as (14)CO(2). Similar cumulative mineralization was obtained when Pantex aquifer material was included in the solution matrix. Other experiments using 4000 mg KMnO(4) L(-1) showed that initial RDX concentrations (1.3-10.4 mg L(-1)) or initial pH (4-11) had little effect on reaction rates. Attempts to identify RDX degradates and reaction products showed that N(2)O was a product of permanganate oxidation and constituted 20 to 30% of the N balance. Time-course measurements of a (14)C-RDX solution treated with KMnO(4) revealed few (14)C-labeled degradates but through liquid chromatography mass spectrometry (LC-MS) analysis, we present evidence that 4-nitro-2,4-diaza butanol is formed. Aquifer microcosm studies confirmed that the transformation products not mineralized by KMnO(4) were much more biodegradable than parent RDX. These results indicate permanganate can effectively transform and mineralize RDX in the presence of aquifer material and support its use as an in situ chemical oxidation treatment for the Pantex perched aquifer. PMID- 15537940 TI - Effects of near-surface hydraulic gradients on nitrate and phosphorus losses in surface runoff. AB - Phosphorous (P) and nitrogen (N) in runoff from agricultural fields are key components of nonpoint-source pollution and can accelerate eutrophication of surface waters. A laboratory study was designed to evaluate effects of near surface hydraulic gradients on P and N losses in surface runoff from soil pans at 5% slope under simulated rainfall. Experimental treatments included three rates of fertilizer input (control [no fertilizer input], low [40 kg P ha(-1), 100 kg N ha(-1)], and high [80 kg P ha(-1), 200 kg N ha(-1)]) and four near-surface hydraulic gradients (free drainage [FD], saturation [Sa], artesian seepage without rain [Sp], and artesian seepage with rain [Sp + R]). Simulated rainfall of 50 mm h(-1) was applied for 90 min. The results showed that near-surface hydraulic gradients have dramatic effects on NO(3)-N and PO(4)-P losses and runoff water quality. Under the low fertilizer treatment, the average concentrations in surface runoff from FD, Sa, Sp, and Sp + R were 0.08, 2.20, 529.5, and 71.8 mg L(-1) for NO(3)-N and 0.11, 0.54, 0.91, and 0.72 mg L(-1) for PO(4)-P, respectively. Similar trends were observed for the concentrations of NO(3)-N and PO(4)-P under the high fertilizer treatment. The total NO(3)-N loss under the FD treatment was only 0.01% of the applied nitrogen, while under the Sp and Sp + R treatments, the total NO(3)-N loss was 11 to 16% of the applied nitrogen. These results show that artesian seepage could make a significant contribution to water quality problems. PMID- 15537941 TI - Development of a phosphorus index for pastures fertilized with poultry litter- factors affecting phosphorus runoff. AB - Currently, several state and federal agencies are proposing upper limits on soil test phosphorus (P), above which animal manures cannot be applied, based on the assumption that high P concentrations in runoff are due to high soil test P. Recent studies show that other factors are more indicative of P concentrations in runoff from areas where manure is being applied. The original P index was developed as an alternative P management tool incorporating factors affecting both the source and transport of P. The objective of this research was to evaluate the effects of multiple variables on P concentrations in runoff water and to construct a P source component of a P index for pastures that incorporates these effects. The evaluated variables were: (i) soil test P, (ii) soluble P in poultry litter, (iii) P in poultry diets, (iv) fertilizer type, and (v) poultry litter application rate. Field studies with simulated rainfall showed that P runoff was affected by the amount of soluble P applied in the fertilizer source. Before manure applications, soil test P was directly related to soluble P concentrations in runoff water. However, soil test P had little effect on P runoff after animal manure was applied. Unlike most other P indices, weighting factors of the P source components in the P index for pastures are based on results from runoff studies conducted under various management scenarios. As a result, weighting factors for the P source potential variables are well justified. A modification of the P index using scientific data should strengthen the ability of the P index concept to evaluate locations and management alternatives for P losses. PMID- 15537942 TI - Evaluation of the phosphorus source component in the phosphorus index for pastures. AB - A phosphorus (P) index for pastures was developed to write nutrient management plans that determine how much P can be applied to a given field. The objectives of this study were to (i) evaluate and compare the P index for pastures, particularly the P source component, and an environmental threshold soil test P level by conducting rainfall simulations on contrasting soils under various management scenarios; and (ii) evaluate the P index for pastures on field-scale watersheds. Poultry litter was applied to 12 small plots on each of six farms based on either an environmental threshold soil test P level or on the P index for pastures, and P runoff was evaluated using rainfall simulators. The P index was also evaluated from two small (0.405 ha) watersheds that had been fertilized annually with poultry litter since 1995. Results from the small plot study showed that soil test P alone was a poor predictor of P concentrations in runoff water following poultry litter applications. The relationship between P in runoff and the amount of soluble P applied was highly significant. Furthermore, P concentrations in runoff from plots with and without litter applications were significantly correlated to P index values. Studies on pastures receiving natural rainfall and annual poultry litter applications indicated that the P index for pastures predicted P loss accurately without calibration (y = 1.16x - 0.23, r(2) = 0.83). These data indicate that the P index for pastures can accurately assess the risk of P loss from fields receiving poultry litter applications in Arkansas and provide a more realistic risk assessment than threshold soil test P levels. PMID- 15537943 TI - Rainfall timing and poultry litter application rate effects on phosphorus loss in surface runoff. AB - Phosphorus (P) in runoff from pastures amended with poultry litter may be a significant contributor to eutrophication of lakes and streams in Georgia and other areas in the southeastern United States. The objectives of this research were to determine the effects of litter application rate and initial runoff timing on the long-term loss of P in runoff from surface-applied poultry litter and to develop equations that predict P loss in runoff under these conditions. Litter application rates of 2, 7, and 13 Mg ha(-1), and three rainfall scenarios applied to 1- x 2-m plots in a 3 x 3 randomized complete block design with three replications. The rainfall scenarios included (i) sufficient rainfall to produce runoff immediately after litter application; (ii) no rainfall for 30 d after litter application; and (iii) small rainfall events every 7 d (5 min at 75 mm h( 1)) for 30 d. Phosphorus loss was greatest from the high litter rate and immediate runoff treatments. Nonlinear regression equations based on the small plot study produced fairly accurate (r(2) = 0.52-0.62) prediction of P concentrations in runoff water from larger (0.75 ha) fields over a 2-yr period. Predicted P concentrations were closest to observed values for events that occurred shortly after litter application, and the relative error in predictions increased with time after litter application. In addition, previously developed equations relating soil test P levels to runoff P concentrations were ineffective in the presence of surface-applied litter. PMID- 15537944 TI - Decreasing phosphorus runoff losses from land-applied poultry litter with dietary modifications and alum addition. AB - Phosphorus (P) losses from pastures fertilized with poultry litter contribute to the degradation of surface water quality in the United States. Dietary modification and manure amendments may reduce potential P runoff losses from pastures. In the current study, broilers were fed a normal diet, phytase diet, high available phosphorus (HAP) corn diet, or HAP corn + phytase diet. Litter treatments were untreated control and alum added at 10% by weight between flocks. Phytase and HAP corn diets reduced litter dissolved P content in poultry litter by 10 and 35%, respectively, compared with the normal diet (789 mg P kg(-1)). Alum treatment of poultry litter reduced the amount of dissolved P by 47%, while a 74% reduction was noted after alum treatment of litter from the HAP corn + phytase diet. The P concentrations in runoff water were highest from plots receiving poultry litter from the normal diet, whereas plots receiving poultry litter from phytase and HAP corn diets had reduced P concentrations. The addition of alum to the various poultry litters reduced P runoff by 52 to 69%; the greatest reduction occurred when alum was used in conjunction with HAP corn and phytase. This study demonstrates the potential added benefits of using dietary modification in conjunction with manure amendments in poultry operations. Integrators and producers should consider the use of phytase, HAP corn, and alum to reduce potential P losses associated with poultry litter application to pastures. PMID- 15537945 TI - Uncertainty assessment of the model RICEWQ in northern Italy. AB - Model predictions are often seriously affected by uncertainties arising from many sources. Ignoring the uncertainty associated with model predictions may result in misleading interpretations when the model is used by a decision-maker for risk assessment. In this paper, an analysis of uncertainty was performed to estimate the uncertainty of model predictions and to screen out crucial variables using a Monte Carlo stochastic approach and a number of statistical methods, including ANOVA and stepwise multiple regression. The model studied was RICEWQ (Version 1.6.1), which was used to forecast pesticide fate in paddy fields. The results demonstrated that the paddy runoff concentration predicted by RICEWQ was in agreement with field measurements and the model can be applied to simulate pesticide fate at field scale. Model uncertainty was acceptable, runoff predictions conformed to a log-normal distribution with a short right tail, and predictions were reliable at field scale due to the narrow spread of uncertainty distribution. The main contribution of input variables to model uncertainty resulted from spatial (sediment-water partition coefficient and mixing depth to allow direct partitioning to bed) and management (time and rate of application) parameters, and weather conditions. Therefore, these crucial parameters should be carefully parameterized or precisely determined in each site-specific paddy field before the application of the model, since small errors of these parameters may induce large uncertainty of model outputs. PMID- 15537946 TI - Water quality impacts of converting to a poultry litter fertilization strategy. AB - When improperly managed, land application of animal manures can harm the environment; however, limited watershed-scale runoff water quality data are available to research and address this issue. The water quality impacts of conversion to poultry litter fertilization on cultivated and pasture watersheds in the Texas Blackland Prairie were evaluated in this three-year study. Edge-of field N and P concentrations and loads in surface runoff from new litter application sites were compared with losses under inorganic fertilization. The impact on downstream nutrient loss was also examined. In the fallow year with no fertilizer application, nutrient losses averaged 3 kg N ha(-1) and 0.9 kg P ha( 1) for the cultivated watersheds and were below 0.1 kg ha(-1) for the pasture watersheds. Following litter application, PO(4)-P concentrations in runoff were positively correlated to litter application rate and Mehlich-3 soil P levels. Following litter application, NO(3)-N and NH(4)-N concentrations in runoff were typically greater from cultivated watersheds, but PO(4)-P concentrations were greater for the pasture watersheds. Total N and P loads from the pasture watersheds (0.2 kg N ha(-1) and 0.7 kg P ha(-1)) were significantly lower than from the cultivated watersheds (32 kg N ha(-1) and 5 kg P ha(-1)) partly due to lower runoff volumes from the pasture watersheds. Downstream N and P concentrations and per-area loads were much lower than from edge-of-field watersheds. Results demonstrate that a properly managed annual litter application (4.5 Mg ha(-1) or less depending on litter N and P content) with supplemental N should supply necessary nutrients without detrimental water quality impacts. PMID- 15537947 TI - Efficacy of vegetated buffer strips for retaining Cryptosporidium parvum. AB - Overland and shallow subsurface hydrologic transport of pathogenic Cryptosporidium parvum oocysts from cattle feces into surface drinking water supplies is a major concern on annual grasslands in California's central and southern Sierra Nevada foothills. Soil boxes (0.5 m wide x 1.1 m long x 0.3 m deep) were used to evaluate the ability of grass vegetated buffer strips to retain 2 x 10(8) spiked C. parvum oocysts in 200-g fecal deposits during simulated rainfall intensities of 30 to 47.5 mm/h over 2 h. Buffers were comprised of Ahwahnee sandy loam (coarse-loamy, mixed, active, thermic Mollic Haploxeralfs; 78:18:4 sand to silt to clay ratio; dry bulk density = 1.4 g/cm(3)) set at 5 to 20% land slope, and >/=95% grass cover (grass stubble height = 10 cm; biomass = 900 kg/ha dry weight). Total number of oocysts discharged from each soil box (combined overland and subsurface flow) during the 120-min simulation ranged from 1.5 x 10(6) to 23.9 x 10(6) oocysts. Observed overall mean log(10) reduction of total C. parvum flux per meter of vegetated buffer was 1.44, 1.19, and 1.18 for buffers at 5, 12, and 20% land slope, respectively. Rainfall application rate (mm/h) was strongly associated with oocyst flux from these vegetated buffers, resulting in a decrease of 2 to 4% in the log(10) reduction per meter buffer for every additional mm/h applied to the soil box. These results support the use of strategically placed vegetated buffers as one of several management strategies that can reduce the risk of waterborne C. parvum attributable to extensive cattle grazing on annual grassland watersheds. PMID- 15537948 TI - Using nitrogen-15 to quantify vegetative buffer effectiveness for sequestering nitrogen in runoff. AB - Previous studies have observed higher levels of soluble nutrients leaving vegetative buffers than entering them, suggesting that the buffers themselves are acting as a source rather than a sink by releasing previously stored nutrients. This study used 98 atom % (15)N-labeled KNO(3) at a rate of 5 kg ha(-1) to quantify buffer efficiency for sequestering new inputs of NO(-)(3)-N in an extensively grazed irrigated pasture system. Buffer treatments consisted of an 8 m buffer, a 16-m buffer, and a nonbuffered control. Regardless of the form of runoff N (NO(-)(3), NH(+)(4), or dissolved organic nitrogen [DON]), more (15)N was lost from the nonbuffered treatments than from the buffered treatments. The majority of the N attenuation was by vegetative uptake. Over the course of the study, the 8-m buffer decreased NO(-)(3)-(15)N load by 28% and the 16-m buffer decreased load by 42%. For NH(+)(4)-(15)N, the decrease was 34 and 48%, and for DON-(15)N, the decrease was 21 and 9%. Although the buffers were effective overall, the majority of the buffer impact occurred in the first four weeks after (15)N application, with the buffered plots attenuating nearly twice as much (15)N as the nonbuffered plots. For the remainder of the study, buffer effect was not as marked; there was a steady release of (15)N, particularly NO(-)(3)- and DON (15)N, from the buffers into the runoff. This suggests that for buffers to be sustainable for N sequestration there is a need to manage buffer vegetation to maximize N demand and retention. PMID- 15537949 TI - Transport of phosphate through artificial macropores during film and pulse flow. AB - Flow through artificial macropores may occur as a water film along the macropore walls (film flow) or as moving water segments separated by air bubbles (pulse flow). To investigate the effect of macropore flow pattern (i.e., film and pulse flow) on the interaction of solutes with macropore walls, we studied orthophosphate (P) transport and sorption in artificial macropores. The experimental setup consisted of a column (height = 20 cm, diameter = 20 cm) homogenously packed with glass beads and fitted at outflow with a vertical artificial macropore placed below the column. The artificial macropore consisted of ceramic tubes (3 or 8 mm i.d.; 31.5 cm long) coated on the inside with iron oxide serving as phosphate sorbents. An orthophosphate solution containing 0.04 mg P L(-1) was applied at a rate of 9 to 12 mm h(-1) to the column, eventually causing macropore flow. In the 8-mm-i.d. tubes only film flow occurred. Pulse flow was dominating in the 3-mm-i.d. tubes. Generally, the flow patterns were reproducible and seldom did pulse flow replaced film flow or vice versa. During film flow, a significantly larger decrease in macropore P concentration per tube was observed relative to that with pulse flow events. However, pulse and film flow lead to almost the same amounts of P sorbed per unit surface area when exposed to the same solute P concentration. Comparison with P sorption capacity experiments indicated that the sorption rate, rather than the sorption capacity, controls the amount of sorbed P during macropore flow in the studied system. PMID- 15537950 TI - Radionuclide transport above a near-surface water table: IV. Soil migration and crop uptake of chlorine-36 and technetium-99, 1990 to 1993. AB - Vertical distributions of (36)Cl and (99)Tc are presented from deep and shallow lysimeters above artificially controlled water tables for a 4-yr experiment from 1990 to 1993. Activity concentration profiles were all measured in late summer when a winter wheat (Triticum aestivum L. cv. Pastiche) crop was harvested. After harvest, activity concentrations in different organs of the crop were determined and crop uptake quantified as both an inventory ratio (IR) and a transfer factor (TF(w)), weighted to account for differential root and radionuclide distributions within the soil profile. Vertical distributions of radionuclides, crop roots within the soil, and IR and TF(w) values were each subjected to analysis of variance to estimate the individual and combined effects of soil depth and the year of the experiment on the results obtained. Chlorine-36 and (99)Tc exhibited highly significant variations in activity concentrations with soil depth and from year to year, indicating considerable physical mobility of both radionuclides. Soil-to-plant transfer was also high for both radionuclides compared with data obtained for gamma-emitting radionuclides. The IR values indicated that up to 40% of (36)Cl was incorporated in the crop's tissues at harvest, compared with a maximum of less than 1% for the less mobile gamma-emitting radionuclides. On the basis of the TF(w) values determined, (36)Cl uptake by winter wheat exceeded (99)Tc uptake, indicating that (36)Cl is highly bioavailable. Factors controlling the migration and bioavailability of both (36)Cl and (99)Tc in soils are discussed. PMID- 15537951 TI - Interrelationship of macropores and subsurface drainage for conservative tracer and pesticide transport. AB - Macropore flow results in the rapid movement of pesticides to subsurface drains, which may be caused in part by a small portion of macropores directly connected to drains. However, current models fail to account for this direct connection. This research investigated the interrelationship between macropore flow and subsurface drainage on conservative solute and pesticide transport using the Root Zone Water Quality Model (RZWQM). Potassium bromide tracer and isoxaflutole, the active ingredient in BALANCE herbicide [(5-cyclopropyl-4-isoxazolyl) [2(methylsulfonyl)-4-(trifluoromethyl)phenyl] methanone], with average half-life of 1.7 d were applied to a 30.4-ha Indiana corn (Zea mays L.) field. Water flow and chemical concentrations emanating from the drains were measured from two samplers. Model predictions of drain flow after minimal calibration reasonably matched observations (slope = 1.03, intercept = 0.01, and R(2) = 0.75). Without direct hydraulic connection of macropores to drains, RZWQM under predicted bromide and isoxaflutole concentration during the first measured peak after application (e.g., observed isoxaflutole concentration was between 1.2 and 1.4 mug L(-1), RZWQM concentration was 0.1 mug L(-1)). This research modified RZWQM to include an express fraction relating the percentage of macropores in direct hydraulic connection to drains. The modified model captured the first measured peak in bromide and isoxaflutole concentrations using an express fraction of 2% (e.g., simulated isoxaflutole concentration increased to 1.7 mug L(-1)). The RZWQM modified to include a macropore express fraction more accurately simulates chemical movement through macropores to subsurface drains. An express fraction is required to match peak concentrations in subsurface drains shortly after chemical applications. PMID- 15537952 TI - Pig slurry application and irrigation effects on nitrate leaching in Mediterranean soil lysimeters. AB - Land application of animal manures, such as pig slurry (PS), is a common practice in intensive-farming agriculture. However, this practice has a pitfall consisting of the loss of nutrients, in particular nitrate, toward water courses. The objective of this study was to evaluate nitrate leaching for three application rates of pig slurry (50, 100, and 200 Mg ha(-1)) and a control treatment of mineral fertilizer (275 kg N ha(-1)) applied to corn grown in 10 drainage lysimeters. The effects of two irrigation regimes (low vs. high irrigation efficiency) were also analyzed. In the first two irrigation events, drainage NO(3)-N concentrations as high as 145 and 69 mg L(-1) were measured in the high and moderate PS rate treatments, respectively, in the low irrigation efficiency treatments. This indicates the fast transformation of the PS ammonium into nitrate and the subsequent leaching of the transformed nitrate. Drainage NO(3)-N concentration and load increased linearly by 0.69 mg NO(3)-N L(-1) and 4.6 kg NO(3)-N ha(-1), respectively, for each 10 kg N ha(-1) applied over the minimum of 275 kg N ha(-1). An increase in irrigation efficiency did not induce a significant increase of leachate concentration and the amount of nitrate leached decreased about 65%. Application of low PS doses before sowing complemented with sidedressing N application and a good irrigation management are the key factors to reduce nitrate contamination of water courses. PMID- 15537953 TI - Phosphorus dynamics in soils receiving chemically treated dairy manure. AB - Chemical treatment of animal manure with Al, Fe, and Ca salts appears capable of concentrating P in a smaller volume, thereby providing increased manure management options. However, little information is available on the fate of nutrients in soils receiving chemically treated manure. An incubation study (1 d to 2 yr) was conducted with three soils (Soils I, II, and III with 12, 66, and 94 mg kg(-1) Bray-1 P, respectively) and four manure treatments (one untreated and three chemical including Al-, Fe-, and Ca-treated) at two rates (12.5 and 25 mg P kg(-1)), and a control (no manure). Subsamples were analyzed for Bray-1 P and water-extractable phosphorus (WEP) after eight incubation time periods. Phosphorus distribution among different fractions (soluble and loosely bound; Al , Fe-, and Ca-bound; organic P; and residual) was also determined after 1 d and 1 yr. Water-extractable P increased when soils received untreated or Ca-treated manure in proportion to P application rate. Water-extractable P, however, decreased (compared with control) for Soils II and III or slightly increased for Soil I with addition of Al- or Fe-treated manure. Water-extractable P decreased sharply between 1 d and 1 to 2 wk and then remained relatively constant or increased slightly up to 2 yr depending on treatment and soil type. Bray-1 P increased for all treatment types and soils in the following order: Ca-treated > Al-treated >/= untreated > Fe-treated > control. Within each treatment, Bray-1 P decreased between 1 d and 1 to 2 wk and then gradually increased for up to 3 mo (Soils II and III) or 6 mo (Soil I). Application of Al- or Fe-treated manure decreased P solubility with the effect being more pronounced in soils with high background P. Since the application of Ca-treated manure increased both WEP and Bray-1 P, it should be recommended for soils where the objective is to increase P availability. Several years of P input through fertilizer and manure contributed mainly to aluminum-bound phosphorus (Al-P) and to a lesser degree to other fractions. Only soluble and loosely bound P (all soils) and Al-P (Soil I) exhibited treatment-type effects after receiving chemically treated manure. The study results will help bridge the gap between our knowledge of chemical treatment systems for animal manure and the ultimate fate of P when the treated manure is land-applied. PMID- 15537954 TI - Influence of phytase addition to poultry diets on phosphorus forms and solubility in litters and amended soils. AB - Diet modification to decrease phosphorus (P) concentration in animal feeds and manures can reduce surpluses of manure P in areas of intensive animal production. We generated turkey and broiler litters from two and three flock trials, respectively, using diets that ranged from "high" to "low" in non-phytate phosphorus (NPP) and some of which contained feed additives such as phytase. Phosphorus forms in selected litters were analyzed by sequential chemical fractionation and solution (31)P nuclear magnetic resonance (NMR) spectroscopy. Selected litters were also incubated with four contrasting soils. Reducing dietary NPP and using phytase decreased total P in litters by up to 38%. Water soluble phosphorus (WSP) in litters was decreased 21 to 44% by feeding NPP closer to animal requirement, but was not affected by phytase addition. Solution (31)P NMR spectroscopy showed that feeding NPP closer to requirement decreased orthophosphate in litters by an average of 38% and that adding phytase to feed did not increase the concentration of orthophosphate in litters. Phytase also decreased phytate P in litters by 25 to 38%, demonstrating that it increases phytate P hydrolysis. Incorporation of litters with soils at the same total P rate increased WSP in soils relative to the control; this increase was correlated to soluble P added with litters at 5 d, but not by 29 d. Changes in soil Mehlich 3 phosphorus (M3-P) were related to total P added in litter, rather than soluble P. We conclude that feeding NPP closer to requirement and using feed additives such as phytase decrease total P concentrations in litters, while having little effect on P solubility in litters and amended soils. PMID- 15537955 TI - Field water balance of landfill final covers. AB - Landfill covers are critical to waste containment, yet field performance of specific cover designs has not been well documented and seldom been compared in side-by-side testing. A study was conducted to assess the ability of landfill final covers to control percolation into underlying waste. Conventional covers employing resistive barriers as well as alternative covers relying on water storage principles were monitored in large (10 x 20 m), instrumented drainage lysimeters over a range of climates at 11 field sites in the United States. Surface runoff was a small fraction of the water balance (0-10%, 4% on average) and was nearly insensitive to the cover slope, cover design, or climate. Lateral drainage from internal drainage layers was also a small fraction of the water balance (0-5.0%, 2.0% on average). Average percolation rates for the conventional covers with composite barriers (geomembrane over fine soil) typically were less than 12 mm/yr (1.4% of precipitation) at humid locations and 1.5 mm/yr (0.4% of precipitation) at arid, semiarid, and subhumid locations. Average percolation rates for conventional covers with soil barriers in humid climates were between 52 and 195 mm/yr (6-17% of precipitation), probably due to preferential flow through defects in the soil barrier. Average percolation rates for alternative covers ranged between 33 and 160 mm/yr (6 and 18% of precipitation) in humid climates and generally less than 2.2 mm/yr (0.4% of precipitation) in arid, semiarid, and subhumid climates. One-half (five) of the alternative covers in arid, semiarid, and subhumid climates transmitted less than 0.1 mm of percolation, but two transmitted much more percolation (26.8 and 52 mm) than anticipated during design. The data collected support conclusions from other studies that detailed, site-specific design procedures are very important for successful performance of alternative landfill covers. PMID- 15537956 TI - Simulating urban waste compost effects on carbon and nitrogen dynamics using a biochemical index. AB - Composting has emerged as a valuable route for the disposal of urban waste, with the prospect of applying composts on arable fields as organic amendments. Proper management of urban waste composts (UWCs) requires a capacity to predict their effects on carbon and nitrogen dynamics in the field, an issue in which simulation models are expected to play a prominent role. However, the parameterization of soil organic amendments within such models generally requires laboratory incubation data. Here, we evaluated the benefit of using a biochemical index based on Van Soest organic matter fractions to parameterize a deterministic model of soil C and N dynamics, NCSOIL, as compared with a standard alternative based on laboratory incubation data. The data included C mineralization and inorganic N dynamics in samples of a silt loam soil (Typic Hapludalf) mixed with various types of UWC and farmyard manure. NCSOIL successfully predicted the various nitrogen mineralization-immobilization patterns observed, but underestimated CO(2) release by 10 to 30% with the less stable amendments. The parameterization based on the biochemical index achieved a prediction error significantly larger than the standard parameterization in only 10% of the tested cases, and provided an acceptable fit to experimental data. The decomposition rates and C to N ratios of compost organic matter varied chiefly according to the type of waste processed. However, 62 to 66% of their variance could be explained by the biochemical index. We thus suggest using the latter to parameterize organic amendments in C and N models as a substitute for time-consuming laboratory incubations. PMID- 15537957 TI - Kinetic effect of humic acid on alachlor degradation by anodic Fenton treatment. AB - Contamination of water often results from the heavy use of agricultural chemicals, and the disposal of aqueous pesticide waste is a concern. Anodic Fenton treatment (AFT) has been shown to be a successful remediation method for pesticides in solution, but the effect of soil on the degradation kinetics of pesticides using this method has not been determined. The purpose of this study was to study the effect of humic acid, as a soil surrogate, on the degradation kinetics of alachlor [2-chloro-N-(2,6-diethylphenyl-N-(methoxymethyl) acetamide], a heavily used herbicide that has been studied in pure aqueous solution by AFT. The AFT consists of a controlled constant delivery of Fenton reagents, using an electrochemical half-cell to deliver ferrous iron. Alachlor was quickly degraded by AFT, and the kinetics were found to obey the previously developed AFT model well. Degradation of alachlor by AFT in humic acid slurry showed that when the amount of humic acid was increased, alachlor degradation was significantly slowed down and the degradation kinetics were shifted from the AFT model to a first order model. Further experimentation indicated that humic acid not only competes with alachlor for hydroxyl radicals, reducing the degradation rate of the target compound, but also buffers the slurry at near neutral pH, blocking regeneration of ferrous ion from ferric ion and subsequently shifting the kinetics to first order. Degradation of several other pesticides in humic acid slurry also followed first-order kinetics. These results imply that higher concentrations of Fenton reagents will be required for soil remediation. PMID- 15537958 TI - Microcosm studies on anaerobic phosphate flux and mineralization of lake sediment organic carbon. AB - Lake sediment has long been recognized as an important source of nutrients such as phosphorus. To gain a better understanding of phosphorus flux at the sediment water interface, it is crucial to investigate the sediment porewater. There is also growing concern and interest in identifying whether organic-rich sediment is an important source of greenhouse gases such as CO(2) and CH(4). In the present study, we took sediment samples from West Lake, a shallow hypereutrophic lake in Hangzhou, Zhejiang Province, China and incubated subsamples under anaerobic conditions at 25 degrees C for 182 d using a specially designed microcosm that permits repeated extraction of sediment porewater and sampling of headspace gases. Anaerobic phosphate fluxes and mineralization of sediment organic carbon were measured. Average diffusive flux of soluble phosphorus was 0.81 mg P m(-2) d(-1) during the initial 18 d of incubation. Decomposition of sediment organic C followed zero-order reaction kinetics and methane accounted for about 50% of the mineralization products. The results suggest that organic-rich sediments can be important sources of P and methane under anaerobic conditions. Laboratory studies simulating field conditions and field studies are necessary to determine the contribution of sediment as a source of P and greenhouse gases. PMID- 15537959 TI - Response of biogeochemical indicators to a drawdown and subsequent reflood. AB - Temporal oscillations in hydrology are a common occurrence in wetlands and can result in alternating flooded and drained conditions in the surface soil. These oscillations in water levels can stimulate microbial activities and result in the mobilization and redistribution of significant amounts of carbon (C), nitrogen (N), and phosphorus (P). The goal of this study was to experimentally simulate a drawdown and reflood of marsh soil from a nutrient-enriched site and a reference site of a wetland (Blue Cypress Marsh Conservation Area, Florida). The goal was to better understand the changes in biogeochemistry and microbial activities present in these soils as a result of hydrological fluctuations. Measurements of dissolved reactive phosphorus (DRP), ammonia, and nitrate in the floodwater indicated significantly higher (alpha = 0.05) NH(4)(+) and DRP fluxes from the nutrient-enriched site; floodwaters in the cores from both sites contained significant NO(3)(-) concentrations (9.6 mg N L(-1)), which was rapidly consumed over the core incubation period (30 d). Water level drawdown and reflooding initially stimulated the soil microbial biomass, methanogenic rates, and extracellular enzyme activities (acid phosphatase and beta-glucosidase). The anaerobic microbial metabolic activities (CO(2)) where initially significantly (alpha = 0.05) enhanced by the reflood, resulting in roughly equivalent rates as the aerobic respiratory activities (CO(2)), presumably as a function of the high water column NO(3)(-) levels. This study illustrates that the reflood event in the hydrological cycles in a wetland can significantly stimulate the activities of hydrolytic enzymes and microbiological communities in these soils. PMID- 15537960 TI - Denitrification in a coastal plain riparian zone contiguous to a heavily loaded swine wastewater spray field. AB - Riparian zones are recognized as landscape features that buffer streams from pollutants, particularly nitrogen. The objectives of this experiment were to (i) assess denitrification activity within a riparian zone and (ii) determine the influence of physical, chemical, and landscape features on denitrification. This experiment was conducted from 1994 to 1997 in North Carolina on a riparian zone contiguous to a spray field that was heavily loaded with swine lagoon wastewater. Denitrification enzyme activity (DEA) was measured on soils collected from (i) the soil surface, (ii) midway between the soil surface and water table, and (iii) above the water table. The DEA ranged from 3 to 1660 microg N(2)O-N kg(-1) soil h(-1). The DEA was highest next to the stream and lowest next to the spray field. Nitrate was found to be the limiting factor for denitrification. The DEA generally decreased with soil depth; means for the surface, middle, and bottom depths were 147, 83, and 67 microg N(2)O-N kg(-1) soil h(-1), respectively. These DEA values are higher than those reported for riparian zones adjoining cropland of the southeastern United States, but are lower than those reported for a constructed wetland used for treatment of swine wastewater. Regression analysis indicated that soil total nitrogen was the highest single factor correlated to DEA (r(2) = 0.65). The inclusion of water table depth, soil depth, and distance from the spray field improved the R(2) to 0.86. This riparian zone possessed sufficient soil area with high denitrifying conditions to be a significant factor in the removal of excess nitrogen in the ground water. PMID- 15537961 TI - Construction of platinum-tipped redox probes for determining soil redox potential. AB - Redox probes are typically constructed by soldering Pt wire to a metal wire or rod, such as copper or brass. The junction between the Pt and wire or rod is often sealed with an epoxy resin and hardener or with heat-shrink tubing. Microcracks (small cracks invisible to the unaided eye) can form in the hardened resin and result in incorrect readings. The hardened resin is not easily removed, making repairs difficult. Heat-shrink tubing is thin, lacks rigidity, and can be damaged in the soil. The method described in this paper used a thick-walled, adhesive-lined terminal insulator to seal the junction. The terminal insulators were easily applied and removed, which made faulty probes easy to repair. Two hundred forty probes were made with this method and eight were made with a marine epoxy resin. The probes were tested with a redox buffer solution (Light Solution) and were usable if they read +476 +/- 10 mV. The probes were installed 0.76 and 1.5 m deep in the soil. The ability of the probes to provide reliable redox readings was examined by testing selected probes after 10 mo of use and testing all of the probes after completion of the study (19 mo). Ten of the twelve probes tested after 10 mo worked satisfactorily, while the other two clearly malfunctioned before testing. After the study was completed, 236 of the 240 of the probes worked satisfactorily. These results indicate that the construction method presented produces reliable, long-lasting probes. PMID- 15537962 TI - Phosphorus composition of manure from swine fed low-phytate grains: evidence for hydrolysis in the animal. AB - Including low-phytic-acid grains in swine diets can reduce P concentrations in manure, but the influence on manure P composition is relatively unknown. To address this we analyzed manure from swine fed one of four barley (Hordeum vulgare L.) varieties. The barley types consisted of wild-type barley (CDC bold, normal barley diet) and three low-phytic-acid mutant barleys that contained similar amounts of total P but less phytic acid. The phytic acid concentrations in the mutant barleys were reduced by 32% (M422), 59% (M635), and 97% (M955) compared with that in the wild-type barley, respectively. Phosphorus concentrations were approximately one-third less in manures from animals fed low phytic-acid barleys compared with those fed the wild-type variety. Phytic acid constituted up to 55% of the P in feed, but only trace concentrations were detected in NaOH-EDTA extracts of all manures by solution (31)P nuclear magnetic resonance (NMR) spectroscopy. Phosphate was the major P fraction in the manures (86-94% extracted P), with small concentrations of pyrophosphate and simple phosphate monoesters also present. The latter originated mainly from the hydrolysis of phospholipids during extraction and analysis. These results suggest that phytic acid is hydrolyzed in swine, possibly in the hind gut by intestinal microflora before being excreted in feces, even though the animals have little phytase activity in the gut and derive little nutritional benefit from phytate P. We conclude that feeding low-phytic-acid grains reduces total manure P concentrations and the manure P is no more soluble than P generated from normal barley diets. PMID- 15537963 TI - Differential diagnosis of polypoid lesions seen at CT colonography (virtual colonoscopy). AB - Computed tomographic (CT) colonography, also referred to as virtual colonoscopy, holds significant promise for effective large-scale colorectal cancer screening. Two-dimensional (2D) and three-dimensional (3D) displays of the CT data are employed, both of which are critical for proper evaluation. Although many radiologists continue to use the 2D images for polyp detection, more emphasis on the 3D images for primary detection of polyps has yielded the best results for screening detection. The primary target lesion for colorectal screening is the adenomatous polyp, since detection and removal of all larger or advanced lesions could potentially prevent approximately 95% or more of all colon cancers. Frankly invasive adenocarcinoma is rarely encountered in an average-risk asymptomatic screening population, but it is of course another important target. In addition to these clinically significant epithelial neoplasms, however, a host of additional lesions and pseudolesions may be encountered that appear polypoid at CT colonography. A subset of "don't touch" lesions, which should not be confused with potential neoplasms, can also be recognized at CT colonography. A variety of useful techniques and observations can be used to increase the specificity of CT colonography for distinguishing false polyps from true polyps. PMID- 15537965 TI - Gastroduodenal stent placement: current status. AB - Gastroduodenal obstruction is a preterminal event in patients with advanced malignancies of the stomach, pancreas, and duodenum. It severely limits the quality of life in affected patients due to constant emesis and associated malnutrition. Surgical gastrojejunostomy has been the traditional palliative treatment but is associated with a high complication rate, and delayed gastric emptying is a frequent problem. Gastroduodenal stent placement is a very safe and effective palliation method in patients with unresectable malignant tumors causing gastric outlet obstruction, with adequate palliation obtained in most cases. The procedure can be performed under fluoroscopic guidance or with a combination of fluoroscopic and endoscopic techniques. Advantages of gastroduodenal stent placement over surgical palliation include suitability as an outpatient procedure, more rapid gastric emptying, greater cost effectiveness, fewer complications, and improved quality of life. Covered duodenal stents are currently being evaluated and may play an increasingly important role in preventing recurrent obstruction secondary to tumor ingrowth. Moreover, simultaneous palliation of biliary and duodenal malignant strictures is possible with the use of metallic stents. Gastroduodenal stent placement is a promising new alternative for the palliation of malignant gastroduodenal obstruction. PMID- 15537966 TI - Unusual causes of tubo-ovarian abscess: CT and MR imaging findings. AB - Actinomycosis, tuberculosis, and xanthogranulomatous inflammation are rare but specific causes of tubo-ovarian abscess (TOA). TOAs with these causes are frequently misdiagnosed as ovarian malignancies due to their unusual appearances at computed tomography (CT) and magnetic resonance (MR) imaging. Tubo-ovarian actinomycosis frequently has a predominantly solid appearance. A linear, solid, well-enhancing lesion extending directly from the mass is a characteristic CT and MR imaging finding. Small rim-enhancing lesions in the solid part of the mass are also suggestive of actinomycosis. Tuberculous TOAs usually mimic peritoneal carcinomatosis from ovarian cancers. The granulomatous and fibrotic nature of this infection may be reflected in the CT and MR imaging appearances, which can help in differentiation. TOAs from xanthogranulomatous inflammation demonstrate more nonspecific imaging findings than actinomycotic or tuberculous TOAs, although multiple xanthogranulomas in the mass may be seen on MR images. Knowledge of these characteristics can help one make the correct diagnosis and treat the patients appropriately. PMID- 15537967 TI - Pelvic arterial hemorrhage in patients with pelvic fractures: detection with contrast-enhanced CT. AB - Arterial hemorrhage is one of the most serious problems associated with pelvic fractures, and it remains the leading cause of death attributable to pelvic fracture. At many trauma centers, contrast material-enhanced computed tomography (CT) is increasingly used for initial diagnosis in the evaluation of patients with pelvic fractures. Extravasation of contrast material in the pelvis at contrast-enhanced CT is an accurate indicator of ongoing arterial hemorrhage in patients with pelvic fractures. Detection of such extravasation on CT scans can lead to prompt performance of angiographic embolization, which can be lifesaving. Furthermore, the site of contrast material extravasation seen at CT corresponds well to the site of bleeding seen at angiography. This correspondence enables the interventional radiologist to selectively study the arteries most likely to be injured and therefore potentially reduce the patient's morbidity and mortality. Knowledge of the relevant pelvic anatomy, including the osseous, ligamentous, and especially axial vascular anatomy, is essential for understanding the relationship between a site of contrast material extravasation at CT and the specific injured artery visualized at angiography. PMID- 15537970 TI - Nursemaid elbow revisited and a review of congenital radioulnar synostosis. PMID- 15537971 TI - Spectrum of thymic uptake at 18F-FDG PET. AB - Fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) has emerged as a strong diagnostic tool for the diagnosis and staging of neoplasms. Uptake in the thymus at (18)F-FDG PET complicates the assessment of mediastinal involvement by tumor in children and young adults. Increased thymic (18)F-FDG uptake may represent normal physiologic uptake but may also indicate the presence of thymic hyperplasia, lymphomatous infiltration, primary thymic neoplasm, or metastatic disease. Familiarity with the patterns of (18)F-FDG uptake that characterize these pathologic conditions is crucial to the interpretation of PET findings in the thymus. In addition, awareness of the subsets of patients in whom physiologic uptake may be seen and of the normal morphologic features and (18)F FDG PET appearance of the thymus, along with a general sense of the upper limits of metabolic activity for physiologic thymic uptake, will aid in differentiating between physiologic thymic uptake and mediastinal disease. In equivocal cases, correlation with morphologic data from computed tomography or magnetic resonance imaging will likely continue to play a key role in diagnosis and will aid in differentiating benign thymic uptake from malignancy. PMID- 15537972 TI - T1 non-small cell lung cancer: imaging and histopathologic findings and their prognostic implications. AB - About 5% of T1 non-small cell lung cancers (ie, lung cancers less than 3 cm in diameter), mostly focal nodular bronchioloalveolar carcinomas and carcinoid tumors, demonstrate no uptake at fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography (PET) and appear to be indolent in growth; consequently, they are associated with long-term patient survival. About 21% of T1 lung cancers show mediastinal nodal metastasis at the time of diagnosis and about 24% show extrathoracic metastasis, mostly brain metastasis, either at the time of diagnosis or at 1-year follow-up. T1 lung cancers with a large ground-glass attenuation component (50% of tumor volume) at thin-section computed tomography (CT) have a good prognosis with less likelihood of mediastinal nodal or extrathoracic metastasis. On the other hand, solid cancer lesions, especially those with a spiculated margin or with bronchovascular bundle thickening in the surrounding lung, more frequently demonstrate local vessel invasion, lymph node metastasis, and extrathoracic metastasis. In these tumors, work-up for extrathoracic metastases (including whole-body FDG PET or brain magnetic resonance imaging and mediastinoscopy for mediastinal nodes) may be needed, even when CT demonstrates no enlarged nodes in the mediastinum. PMID- 15537974 TI - Treatment of intracranial dural arteriovenous fistulas: current strategies based on location and hemodynamics, and alternative techniques of transcatheter embolization. AB - Intracranial dural arteriovenous fistulas (AVFs) can occur anywhere within the dura mater. Patients may be clinically asymptomatic or may experience symptoms ranging from mild symptoms to fatal hemorrhage, depending on the location (eg, cavernous sinus, transverse-sigmoid sinus, tentorium, superior sagittal sinus, anterior fossa) and venous drainage pattern of the AVF. In the past, dural AVFs have been treated with a variety of approaches, including surgical resection, venous clipping, transcatheter embolization, radiation therapy, or a combination of these treatments. Recent developments in catheter intervention now allow most patients to be cured with transcatheter embolization, although stereotactic radiation therapy is demonstrating good results in an increasing number of cases and surgery is still the preferred option in some cases. Familiarity with drainage patterns, the risk of aggressive symptoms, recent technical advances, and current treatment strategies is essential for the treatment of intracranial dural AVFs. PMID- 15537975 TI - Lumps and bumps on the head in children: use of CT and MR imaging in solving the clinical diagnostic dilemma. AB - Lumps and bumps of the scalp are a common presenting complaint in children and often pose a diagnostic dilemma. These lesions can be difficult to image, with evaluation confounded by their small size. However, accuracy in diagnosis is critical because the diagnostic and therapeutic implications can vary significantly. The clinical examination can be helpful in developing the differential diagnosis and the imaging strategy. Often, however, a single imaging study is insufficient, and the radiologist finds it necessary to image with more than one modality to correctly diagnose a lesion and provide adequate information for the surgeon. Radiography and ultrasonography are often the initial screening diagnostic tests, followed by magnetic resonance (MR) imaging or computed tomography (CT) for more detail. Multidetector thin-section CT and thin-section MR imaging with surface coils are beneficial in the work-up of these small lesions of the head and neck. The use of newer MR imaging sequences such as heavily T2-weighted single-shot turbo spin-echo imaging and diffusion-weighted imaging can improve the characterization of difficult lesions. Familiarity with the variety of new imaging tools and techniques that are available can help characterize pediatric head and neck lesions and guide clinical management. PMID- 15537976 TI - Artifacts in CT: recognition and avoidance. AB - Artifacts can seriously degrade the quality of computed tomographic (CT) images, sometimes to the point of making them diagnostically unusable. To optimize image quality, it is necessary to understand why artifacts occur and how they can be prevented or suppressed. CT artifacts originate from a range of sources. Physics based artifacts result from the physical processes involved in the acquisition of CT data. Patient-based artifacts are caused by such factors as patient movement or the presence of metallic materials in or on the patient. Scanner-based artifacts result from imperfections in scanner function. Helical and multisection technique artifacts are produced by the image reconstruction process. Design features incorporated into modern CT scanners minimize some types of artifacts, and some can be partially corrected by the scanner software. However, in many instances, careful patient positioning and optimum selection of scanning parameters are the most important factors in avoiding CT artifacts. PMID- 15537977 TI - From the archives of the AFIP: pilocytic astrocytoma: radiologic-pathologic correlation. AB - Pilocytic astrocytoma is the most common pediatric central nervous system glial neoplasm and the most common pediatric cerebellar tumor. This tumor has a noteworthy benign biologic behavior that translates into an extremely high survival rate-94% at 10 years-that is by far the best of any glial tumor. Most patients present in the first 2 decades, and clinical symptoms and signs are usually of several months duration and directly related to the specific location of the tumor. The cerebellum, optic nerve and chiasm, and hypothalamic region are the most common locations, but the tumor can also be found in the cerebral hemisphere, ventricles, and spinal cord. Surgical resection is the treatment of choice for all tumors, except for those involving the optic pathway and hypothalamic region, which may be treated with radiation therapy and chemotherapy. Cross-sectional imaging often demonstrates a classic appearance: a cystic mass with an enhancing mural nodule. Less common appearances are quite nonspecific. Surrounding vasogenic edema is rarely present, and this feature provides a valuable clue to the correct diagnosis. Accurate interpretation of imaging studies plays an essential role in directing treatment of these tumors, particularly when they arise in the optic pathway of patients with neurofibromatosis type 1. Disseminated disease and recurrence are extremely rare. PMID- 15537978 TI - Best cases from the AFIP: anaplastic wilms tumor: radiologic and pathologic findings. PMID- 15537979 TI - Metastatic melanoma: an unusual diagnosis for a large anterior mediastinal mass. PMID- 15537980 TI - Pediatric hepatic hemangioma. PMID- 15537981 TI - Medical devices of the chest. AB - Chest devices are encountered on a daily basis by almost all radiologists. A multitude of extrathoracic materials, from intravenous catheters to oxygen tubing and electrocardiographic leads, frequently overlie the chest, neck, and abdomen. Chest tubes, central venous catheters, endotracheal tubes, and feeding tubes are very common. Cardiac surgery involves the use of many sophisticated devices and procedures, ranging from valve replacement to repair of complex congenital anomalies. Coronary artery bypass surgery is no longer considered unusual, and in many large medical centers, ventricular assist devices and total artificial hearts are frequently encountered. Breast implants are visible at standard chest radiography, and many ancillary devices not intended for treatment of cardiac or thoracic diseases are visible on chest radiographs. New devices are constantly being introduced, but most of them are variations on a previous theme. Knowing the specific name of a device is not important. It is important to recognize the presence of a device and to have an understanding of its function, as well as to recognize the complications associated with its use. PMID- 15537982 TI - AAPM/RSNA physics tutorial for residents: digital mammography: an overview. AB - Recent advances in digital detector technology have paved the way to full-field digital mammography (FFDM) systems. The performance of these systems has evolved to the point where replacement of screen-film mammography (SFM) systems is becoming realistic. Despite some commonality between the two techniques, there are fundamental differences in how images are recorded, displayed, and stored. These differences necessitate an understanding of the principles of detection and the characteristics of digital images. Several approaches have been taken in the development of FFDM systems: (a) slot scanning with a scintillator and a charge coupled device (CCD) array, (b) a flat-panel scintillator and an amorphous silicon diode array, (c) a flat-panel amorphous selenium array, (d) a tiled scintillator with fiberoptic tapers and a CCD array, and (e) photostimulable phosphor plates (computed radiography). Although the initial cost of an FFDM system is high compared with that of an SFM system, digital mammography has inherent advantages, such as wide dynamic range, reduction in recall rates, potential for reduction in radiation dose, increased patient throughput, postprocessing capability, and digital acquisition. These advantages and the rapidly occurring technologic developments will help establish FFDM as a mainstay of breast evaluation. PMID- 15537983 TI - RadioGraphics, informatics, bill G, and open source. PMID- 15537984 TI - Informatics in radiology (infoRAD): free DICOM image viewing and processing software for the Macintosh computer: what's available and what it can do for you. AB - It is often necessary for radiologists to use digital images in presentations and conferences. Most imaging modalities produce images in the Digital Imaging and Communications in Medicine (DICOM) format. The image files tend to be large and thus cannot be directly imported into most presentation software, such as Microsoft PowerPoint; the large files also consume storage space. There are many free programs that allow viewing and processing of these files on a personal computer, including conversion to more common file formats such as the Joint Photographic Experts Group (JPEG) format. Free DICOM image viewing and processing software for computers running on the Microsoft Windows operating system has already been evaluated. However, many people use the Macintosh (Apple Computer) platform, and a number of programs are available for these users. The World Wide Web was searched for free DICOM image viewing or processing software that was designed for the Macintosh platform or is written in Java and is therefore platform independent. The features of these programs and their usability were evaluated. There are many free programs for the Macintosh platform that enable viewing and processing of DICOM images. PMID- 15537985 TI - Informatics in radiology (infoRAD): NeatVision: visual programming for computer aided diagnostic applications. AB - A free visual programming-based image analysis development environment for medical imaging applications called NeatVision was developed to provide high level access to a wide range of image processing algorithms through a well defined, easy-to-use graphical interface. The system contains over 300 image manipulation, processing, and analysis algorithms. For more advanced users, an upgrade path is provided to extend the core library with use of the developer's interface, giving users access to additional plug-in features, automatic source code generation, compilation with full error feedback, and dynamic algorithm updates. NeatVision was designed to allow users at all levels of expertise to focus on the computer vision design task for computer-aided diagnostic (CAD) applications rather than the subtleties of a particular programming language. The environment allows the designers of image analysis-based CAD techniques to implement their ideas in a dynamic and straightforward manner. Both NeatVision standard and developer's versions can be downloaded free of charge from the Internet and can run on a variety of computer platforms. PMID- 15537986 TI - The functional architecture of human empathy. AB - Empathy accounts for the naturally occurring subjective experience of similarity between the feelings expressed by self and others without loosing sight of whose feelings belong to whom. Empathy involves not only the affective experience of the other person's actual or inferred emotional state but also some minimal recognition and understanding of another's emotional state. In light of multiple levels of analysis ranging from developmental psychology, social psychology, cognitive neuroscience, and clinical neuropsychology, this article proposes a model of empathy that involves parallel and distributed processing in a number of dissociable computational mechanisms. Shared neural representations, self awareness, mental flexibility, and emotion regulation constitute the basic macrocomponents of empathy, which are underpinned by specific neural systems. This functional model may be used to make specific predictions about the various empathy deficits that can be encountered in different forms of social and neurological disorders. PMID- 15537987 TI - The neurobiology of category learning. AB - Many recent studies have examined the neural basis of category learning. Behavioral neuroscience results suggest that both the prefrontal cortex and the basal ganglia play important category-learning roles; neurons that develop category-specific firing properties are found in both regions, and lesions to both areas cause category-learning deficits. Similar studies indicate that the inferotemporal cortex does not mediate the learning of new categories. The cognitive neuroscience literature on category learning appears contradictory until the results are partitioned according to the type of category-learning task that was used. Three major tasks can be identified: rule based, information integration, and prototype-distortion. Recent results are consistent with the hypotheses that (a) learning in rule-based tasks requires working memory and executive attention and is mediated by frontal-striatal circuits, (b) learning in information-integration tasks requires procedural memory and is mediated primarily within the basal ganglia, and (c) learning in prototype-distortion tasks depends on multiple memory systems, including the perceptual representation system. PMID- 15537988 TI - Psychoneuroimmunology: then and now. AB - Psychoneuroimmunology (PNI) emerged in the neurosciences in the late 1970s to early 1980s and has extended to influence the fields of psychology, psychiatry, endocrinology, physiology, and the biomedical research community. This review documents the journey of PNI from the early 1980s to the present. Today, we recognize that the highly complex immune system interacts with an equally complex nervous system in a bidirectional manner. Evolutionarily old signals continue to play a role in these communications, as do mechanisms for protection of the host. The disparity between physical and psychological stressors is only an illusion. Host defense mechanisms respond in adaptive and meaningful ways to both. The present review will describe a new way of thinking about evolutionarily old molecules, heat shock proteins, adding to a body of evidence suggesting that activation of the acute stress response is a double-edged sword that can both benefit and derail optimal immunity. PMID- 15537989 TI - A central role for norepinephrine in the modulation of cerebellar learning tasks. AB - Norepinephrine (NE) is a central nervous system neuromodulator that enhances the actions of other neurotransmitters such as gamma-aminobutyric acid and glutamate. Based on the Marr-Albus theories, Gilbert suggested that NE influences consolidation of cerebellar learning. NE depletion or blockade of postsynaptic noradrenergic receptors decreases the rate of learning in several cerebellar dependent learning tasks. Loss of cerebellar beta-adrenergic receptor function correlates with a loss of function in related learning tasks. Interventions that improve beta-adrenergic receptor function also improve performance in cerebellum dependent learning tasks. Thus, the authors propose that NE has a central role in the modulation of learning within the cerebellum. PMID- 15537991 TI - The role of technology in task-oriented training in persons with subacute stroke: a randomized controlled trial. AB - This trial compares the effects of task-oriented physical therapy (PT) provided with and without the use of rehabilitation technology on locomotor recovery in 63 persons with subacute stroke. Participants in the experimental (EXP) group used a treadmill, a Kinetron isokinetic exerciser, and a limb-load monitor, whereas those in the control (CTL) group did not while engaging in PT 1 h per day, 5 days per week for 2 months. Locomotor recovery was assessed by clinical (gait speed, Fugl Meyer motor leg and arm subscores, the Balance Scale, the Timed Up and Go, and the Barthel ambulation subscore) and laboratory outcomes (gait kinematics and kinetics) pre- and posttherapy and 3 months later. Within groups, gait speed (P < 0.01) and all secondary measures improved posttherapy (P < 0.01-0.05), and improvements in clinical measures were maintained at follow-up, but there was no difference between groups (P > 0.05). When the groups were pooled, the increase in gait speed was associated (r = 0.52, P = 0.003) with an increase in ankle power generation of the affected leg. The results demonstrate that the efficacy of the task-oriented approach is not dependent on rehabilitation technology. PMID- 15537992 TI - Effects of transcutaneous electrical nerve stimulation (TENS) on cognition, behavior, and the rest-activity rhythm in children with attention deficit hyperactivity disorder, combined type. AB - OBJECTIVE: The aim of this study was to examine the effects of transcutaneous electrical nerve stimulation (TENS) on cognition, behavior, and the rest-activity rhythm in children with attention deficit hyperactivity disorder, combined type (ADHD-CT). METHODS: Twenty-two children diagnosed with ADHD-CT received TENS treatment during 6 weeks, 2 times 30 min a day. Neuropsychological tests were administered to assess cognition, parent/teacher behavioral rating scales were used to measure behavior, and actigraphy was used to assess the rest-activity rhythm. RESULTS: TENS appeared to have a moderate beneficial influence on cognitive functions that load particularly on executive function. There was also improvement in behavior as measured by parent/teacher behavioral rating scales. Moreover, motor restlessness during sleep and motor activity during the day decreased by TENS. CONCLUSIONS: The effects of TENS in children with ADHD are modest but encouraging and warrant further research. PMID- 15537993 TI - Contribution of muscle strength and integration of afferent input to postural instability in persons with stroke. AB - OBJECTIVE: To determine the relationship of muscle strength to postural sway in persons with stroke under standing conditions in which vision and ankle proprioception were manipulated. METHODS: Forty persons with stroke and 40 healthy older adult controls were recruited from the community and underwent balance testing consisting of 6 conditions that manipulate vision and somatosensory information while standing. Postural sway was measured during each condition. In addition, lower extremity joint torques and cutaneous sensation from the plantar surface of the foot were assessed. RESULTS: Postural sway was increased with more challenging standing conditions (i.e., when multiple sensory systems were manipulated) to a greater extent with the group with stroke compared to controls. Muscle strength was only correlated to sway during the most challenging conditions. Furthermore, a greater number of persons with stroke fell during the balance testing compared to controls. CONCLUSIONS: Impairments in re weighting/integrating afferent information, in addition to muscle weakness, appear to contribute to postural instability and falls in persons with stroke. These findings can be used by clinicians to design effective interventions for improving postural control following stroke. PMID- 15537994 TI - Immediate and long-term changes in corticomotor output in response to rehabilitation: correlation with functional improvements in chronic stroke. AB - The goal of this study was to determine whether transcranial magnetic stimulation (TMS) measures of the corticomotor pathways might reflect and predict functional improvements during rehabilitation of a hemiparetic upper extremity. Ten patients with variable levels of functional impairment in upper extremity use were enrolled at least 3 months after a stroke. TMS was used to obtain serial measures of motor thresholds and motor-evoked potential (MEP) size for a muscle from both hands before and after each session of an intervention aimed at improving functional motor control. Functional ability and cortical map area and volume were measured before each therapy session. At intake, all TMS measures from the affected side were impaired compared with the unaffected side but they did not reliably predict the level of functional improvement. Motor thresholds decreased, whereas MEP amplitude and map size increased with treatment. The amount of change in affected side measures was correlated with the amount of improvement in hand/arm function. Normalization of MEP amplitude asymmetry in response to the 1st 2 therapy sessions predicted long-term improvement in Fugl-Meyer motor score. Within limits, TMS appears to be useful in both moderate and more impaired patients as a physiological assay of treatment-induced plasticity and behavioral gains. Methodological differences in the literature, however, currently obscure a full understanding of the potential contributions of TMS to rehabilitation research. PMID- 15537995 TI - The effects of constraint-induced therapy on precision grip: a preliminary study. AB - OBJECTIVE: This preliminary study examines the effects of a 2-week constraint induced therapy (CIT) intervention on the force-producing capabilities of the hemiparetic hand during the performance of a functional dexterous manipulation task. METHODS: A 6-degree-of-freedom force/torque transducer that was embedded into the handle of a key allowed for the quantification of grasping forces and torques produced during the performance of a functional key-turning task. Clinical and kinetic data were collected from 10 subacute patients (3-9 months poststroke) who were participating in an ongoing national clinical study (EXCITE trial) examining the effects of CIT on upper extremity motor performance. Investigators were blinded to treatment designation. Five patients receiving treatment immediately completed 2 weeks of intensive CIT, whereas a group randomized to treatment 1 year later did not receive any therapy during a similar 2-week span. RESULTS: . Results indicated that 4 of the 5 patients in the CIT group, compared to the delayed group, showed significant clinical improvements in hand function, increased maximum precision grip force, improved force and torque regulation, and reduced variability in rate of force production during task performance. CONCLUSIONS: Improved force control may be a mechanism contributing to the observed improvements in dexterous function in those patients undergoing CIT. PMID- 15537996 TI - Effects of acupuncture treatment on poststroke motor recovery and physical function: a pilot study. AB - This pilot study obtained preliminary data on the effects of acupuncture treatment combined with a standard inpatient stroke rehabilitation program on poststroke motor recovery and physical function. Thirty-two patients with acute stroke were recruited and randomized to 1 of 2 treatment arms: standard rehabilitation (control group) or a combination of acupuncture and standard rehabilitation (acupuncture group). Baseline and discharge assessments were obtained on motor recovery as measured by the Fugl-Meyer (FM) Assessment and on physical function as measured by the Functional Independence Measure (FIM). Comparisons were made between the acupuncture and control group in total FM and FIM as well as for each subscale of the FM and FIM. No differences between treatment groups were found in the total FM or the total FIM. However, statistically significant benefit due to acupuncture was observed for the FM lower extremity motor function subscale (P = 0.01) and the tub/shower transfer mobility subscale of the FIM (P = 0.03). Marginally significant benefit due to acupuncture was noted for the toilet transfer mobility subscale of the FIM (P = 0.09). The effectiveness of acupuncture as an adjunct to standard poststroke rehabilitation programs may be demonstrated when more specific measures of stroke motor recovery and physical function are used. PMID- 15537997 TI - The use of hydrotherapy for the management of spasticity. AB - INTRODUCTION: Spasticity is a major problem for the rehabilitation team. Physiotherapy is a vital component of therapy. Oral medication and other modalities such as heat, cold, ultrasound, electrical stimulation, and surgery (neuro-surgical or orthopedic) can also be used. The aim of this study was to compare the effects of hydrotherapy on spasticity and Functional Independence Measure (FIM) scores of patients with spinal cord injury (SCI). MATERIALS AND METHODS: This is a control case matched study. Twenty SCI patients were divided into 2 groups and matched for age, gender, injury time, Ashworth scores, oral baclofen intake, American Spinal Injury Association, and FIM scores. The control group received passive range of motion exercise twice a day and oral baclofen for 10 weeks. The study group also received passive range of motion and oral baclofen, as well as 20 min of water exercises (at 71 degrees F, full immersion) 3 times per week. The authors evaluated spasm severity, FIM scores, oral baclofen intake, and Ashworth scales, between groups at the beginning and at the end of the treatment period. RESULTS: Both groups demonstrated a significant increase in FIM scores. However, the hydrotherapy group demonstrated a larger increase (P < 0.0001) than the control group. There was a statistically significant decrease in oral baclofen intake in the hydrotherapy group (P < 0.01). There was no statistical change in the control group. Spasticity was evaluated by the Ashworth scale. There was a statistical improvement in each group (P < 0.01, P < 0.02). However, when compared to the control group, the use of hydrotherapy produced a significant decrease in spasm severity (P < 0.02). CONCLUSION: Side effects are often seen when using oral drug treatment for spasticity. Adding hydrotherapy to the rehabilitation program can be helpful in decreasing the amount of medication required. Future studies must evaluate benefits of hydrotherapy for rehabilitation. PMID- 15538002 TI - Qualitative comparison: appropriateness, equivalence, and fit. PMID- 15538003 TI - The narrative correspondence method: what a follow-up study can tell us about the longer term effect on participants in emotionally demanding research. AB - The author has based this article on qualitative data gathered from the parents of young adults with cancer who, over the previous 4 years, had contributed narrative accounts to a research project on the experience of caring for a young adult son or daughter with the illness. In the follow-up study, she sought to understand the longer term effect of research participation on the parents. Results show that the parents valued their involvement, and many had found the process of writing therapeutic. Their feelings of isolation had been reduced, and overall, they believed they had benefited from contributing. However, there is also an acknowledgment of the emotional demands caused by recalling painful memories for research purposes. Participants experienced the outcomes of the research as significant and interpreted resulting publications as a lasting memorial to their son or daughter. PMID- 15538004 TI - Qualitative metasynthesis: reflections on methodological orientation and ideological agenda. AB - In an era of pressure toward evidence-based health care, we are witnessing a new enthusiasm for qualitative metasynthesis as an enterprise distinct from conventional literature reviews, secondary analyses, and the many other scholarly endeavors with which it is sometimes confused. This article represents the reflections of five scholars, each ofwhom has authored a distinct qualitative metasynthesis strategy. By providing the reader a glimpse into the tradition of their various qualitative metasynthesis projects, these authors offer a finely nuanced examination of the tensions between comparison and integration, deconstruction and synthesis, and reporting and integration within the metasynthesis endeavor. In so doing, they account for many of the current confusions about representation and generalization within the products of these inquiries. Through understanding the bases of their unique angles of vision, the reader is invited to engage in their commitment to scholarly integrity and intellectual credibility in this emerging methodological challenge. PMID- 15538005 TI - Using qualitative research. AB - A renewed urgency has emerged in the qualitative health research community concerning the utility of qualitative research. This urgency is the result of several converging trends in health care research, including the elevation of practical over basic knowledge, proliferation of qualitative health research studies, and the rise of evidence-based practice as a paradigm and methodology for health care. Diverse conceptualizations of use and users exist, and these have different implications for understanding, demonstrating, and enhancing the utility of qualitative research findings. Issues affecting the utilization of these findings include the varied ways in which they are conceived, presented, synthesized, signified, and translated, and the complex repertoire of skills required to activate the knowledge transformation cycle in qualitative health research fully. PMID- 15538006 TI - Constructing qualitatively derived theory: concept construction and concept typologies. AB - Although concepts differ in scope, specificity, and function within qualitatively derived theory (QDT), and the organization and integration of concepts is essential for the attainment of theoretical integrity, this topic has not been discussed previously in the literature. In this presentation, the author discusses the derivation and the kinds of concepts that qualitative inquiry generates. She examines the various positioning of certain types of concepts in emerging theoretical schemes and how the contribution of those concepts to completed theory varies according to the researchers' agenda and the various roles assumed by different types of concepts. PMID- 15538007 TI - Using a matrix in life transition research. AB - Life transitions can be planned or can occur unexpectedly. They can cause a major change to a person's life patterns and well-being. Older adulthood is a time for many life transitions as a result of changes in life roles and health status. In this exploratory study, the authors investigate the transition involved in driving cessation for older people. In analyzing and organizing the data, they develop a matrix that incorporated descriptive and temporal factors associated with the transition. This matrix is useful in organizing and communicating the findings as a whole and could be used in describing individual experiences. It might be of use for the organization of qualitative data about other life transitions such as illness, retirement, and the development and adoption of new behaviors. PMID- 15538008 TI - Practical issues in using a card sort in a study of non-support and family caregiving. AB - The authors successfully used the card sort data collection technique with 17 female family caregivers in a large ethnographic study of non-support. In this article, they describe the practical issues they addressed. Initially, they developed strategies to construct meaningful statements that reflected key themes and were manageable in an interview. Later, to address challenges for participants, they developed approaches to assist women in moving beyond their own experience, dealing with test anxiety, and anticipating an emotional response. To facilitate effective data collection, they made detailed arrangements in advance, collected "talk aloud" data that captured women's decisions, and maintained accurate records. The women felt validated in their caregiving roles, as the card statements reflected their experience and rich data was elicited. The card sort exercise contributed information about variability in the meaning of similar interactions for different women and a beginning understanding of the criteria women used to make decisions. PMID- 15538009 TI - Bracketing in research: a typology. AB - The term bracketing has increasingly been employed in qualitative research. Although this term proliferates in scientific studies and professional journals, its application and operationalization remains vague and, often, superficial. The growing disconnection of the practice of bracketing in research from its origins in phenomenology has resulted in its frequent reduction to a formless technique, value stance, or black-box term. Mapping the subtle theoretical and philosophical underpinnings of bracketing will facilitate identification and delineation of core elements that compose bracketing, and distinguish how different research approaches prioritize different bracketing elements. The author outlines a typology of six distinct forms of bracketing that encompasses the methodological rigor and evolution of bracketing within the richness of qualitative research. PMID- 15538013 TI - Low health literacy: a unique challenge for HIV/AIDS nursing. PMID- 15538014 TI - New York University Medical Center's pilot "Epo-Depot" program: a win-win for patients and health care. AB - Therapy associated with treatment-related anemia continues to be of great concern for health care providers. Patient satisfaction, patient adherence, and growing insurance reimbursement concerns related to Epoetin alpha therapy for anemia initiated the development of a nursing-led process improvement team at New York University (NYU) Medical Center. Multiple patient groups were identified and treated for anemia with Epoetin alpha through the guide of a standardized pathway providing a convenient and effective treatment plan. The treatment delivery had to be consistent with institutional reimbursement requirements while avoiding significant cost to the patient. Patient populations diagnosed with HIV anemia, HIV oncology, presurgical anemia, and medically complex anemia can now receive appropriate ambulatory treatment. This article describes the formation of the "Epo-Depot" and the logistics involved to service these patients with anemia. PMID- 15538015 TI - Potential mechanisms for altered iron metabolism in human immunodeficiency virus disease. AB - Recently, anemia associated with human immunodeficiency virus (HIV) disease has received more attention as our understanding of the significance of anemia in this population has grown and more emphasis is placed on the quality of life of people living with HIV/AIDS. Although the diagnosis and treatment of anemia in HIV disease has been discussed in great detail, the prevalence and pathophysiology of the two most common forms of anemia, iron deficiency anemia (IDA) and the anemia of chronic disease (ACD), have not received much attention despite the difficulty and importance of differentiating between these two anemias. In addition, little attention has been given to iron overload, which has serious implications in individuals with HIV disease. This article proposes a model of altered iron metabolism in HIV disease as a basis for explaining the pathophysiology and implications of IDA, ACD, and iron overload in this population. Implications for clinical practice and recommendations for future research are discussed. PMID- 15538016 TI - Lipodystrophy syndrome: the morphologic and metabolic effects of antiretroviral therapy in HIV infection. AB - Widespread use of highly active antiretroviral therapy (HAART) to manage HIV infection is now associated with the development of lipodystrophy syndrome. This syndrome is a combination of such morphologic and metabolic changes as hyperlipidemia, fat redistribution, and insulin resistance. Although many of the long-term effects of HAART have not been fully recognized, it is thought that lipodystrophy syndrome may now contribute to early-onset hypercholesterolemia, heart disease, and diabetes, and may have a negative psychological impact on the individual living with HIV infection. PMID- 15538017 TI - Enfuvirtide (T-20): potentials and challenges. AB - Enfuvirtide is a fusion inhibitor used for the treatment of HIV infection. It is an injectable drug, with the patient being responsible for reconstitution as well as injection. The authors present the results of Phase III trials with enfuvirtide. A summary of efficacy, tolerability, pharmacokinetics, resistance, and adverse events from the Phase III trials with enfuvirtide is presented. Enfuvirtide is well tolerated, and no limiting pharmacokinetic interactions have been published thus far. A review of patient perception of self-injection is also summarized. The data reviewed demonstrate that most patients find self-injection easy and do not find that it interferes with activities of daily living. Data on injection site reactions (ISRs), one of the most common adverse events of enfuvirtide, is highlighted, as is nursing management of ISRs. Finally, the authors discuss research that could help further understanding of enfuvirtide and examine some barriers clinicians may face when prescribing this medication. PMID- 15538018 TI - Cryotherapy for treatment of anogenital warts. AB - Human papillomavirus is a group of viruses that produce anogenital warts for which there is no cure. Untreated anogenital warts can spread and form large clusters that are bothersome, painful, and embarrassing to the patient as well as being a significant health risk. One of the infectious disease clinics of the authors' hospital sees an average of 200 HIV-positive individuals per month. Approximately 10% of these patients have anogenital warts. The attending physicians asked the clinical nursing staff if they would be willing to be trained to remove these warts using cryotherapy techniques, if patient efforts at other remedies failed. This request provoked a decision-making challenge for the staff. This article will describe the decision-making process and conclude with the procedure that was created as a result of the decision making. PMID- 15538019 TI - Funding a research project. PMID- 15538022 TI - Adult antisocial behavior following childhood abuse: a new protective factor? PMID- 15538023 TI - Sexual predator civil commitment: a comparison of selected and released offenders. AB - This study compared two groups of sex offenders who were considered for civil commitment under Florida's Jimmy Ryce Act: Two hundred twenty-nine sex offenders who were recommended by forensic evaluators to be civilly committed and 221 sex offenders who were recommended for release. It was hypothesized that selected offenders would be more likely to display risk factors for sex offense recidivism than those who did not meet criteria. Data analyses revealed that selected offenders, as a group, scored significantly higher on actuarial risk assessment instruments. There were also significant differences between the groups on other risk factors that have been empirically correlated with sexual recidivism. Selected offenders had higher frequencies of paraphilia diagnoses and antisocial personality. These findings supported the hypotheses and suggested that evaluators are correctly selecting for civil commitment those sex offenders who have a mental abnormality predisposing them to sexual violence and who are at higher risk for reoffense. PMID- 15538024 TI - The relationship between malingerers' intelligence and MMPI-2 knowledge and their ability to avoid detection. AB - One of the most frequently administered psychometrics is the Minnesota Multiphasic Personality Inventory-2 (MMPI-2). Occasionally, those participants taking the MMPI-2 will malinger or exaggerate their symptoms. Several malingering detection devices are available, and a significant body of literature exists concerning their efficacy. However, little research is available considering those factors that facilitate successfully evading detection as a malingerer. Some of these studies have identified general intelligence and knowledge of the MMPI-2 as key variables in the likelihood of escaping detection as a malingerer. The extant research considered the utility of general intelligence and knowledge of the MMPI-2 as predictors in avoiding detection as a malingerer. To detect malingering, the two traditional detection devices were employed: the F-Scale and the F - K Index. Results indicate that intelligence and MMPI-2 knowledge contribute significantly to the likelihood of successfully escaping detection as a malingerer. PMID- 15538025 TI - Exploring the forensic use of the emotional recognition test (ERT). AB - General "hypo-emotionality" has now become a broadly accepted concept as one of the conspicuous psychological characteristics of repetitive, violent offenders. Numerous psychophysiological studies have verified this premise. The current study sought to examine the applicability of the Emotional Recognition Test (ERT) in a sample of 85 Korean inmates in a maximum-security prison. Two additional criterion groups were included for comparison: normal controls and schizophrenic inpatients. Because the ERT consists of problem-solving items, it is relatively free from response biases and is more efficient than measuring physiological responses. Results indicated that all criterion groups scored significantly differently on the ERT total and subscale scores. Additionally, similar to the physiological emotion-specific findings, emotion-specific subscales of the ERT indicated that offenders had significantly more difficulty in recognizing negative emotional stimuli, particularly sadness. PMID- 15538026 TI - Childhood sexual abuse, gender, and depression among incarcerated youth. AB - This study examined whether sexual abuse was related to higher levels of psychological distress among incarcerated youth and if the effects were invariant across gender. Participants were male and female adolescents, aged 11 to 20 years, incarcerated in six juvenile correctional institutions in five states. Using data from a national study of juveniles confined to secure institutions, this study assessed the relationship between gender, sexual abuse, and depression. Adolescents who were sexually abused during childhood compared to adolescents who were not report higher levels of depression. Sexually abused girls and boys report higher levels of depression compared to boys not sexually abused. These findings suggest the importance of understanding the relationship between childhood sexual abuse and depressive symptoms among incarcerated youth. Additionally, the findings suggest that the effects of childhood sexual abuse are similar for males and females among a high-risk sample of institutionalized juvenile delinquents. Implications for treatment of incarcerated youth are discussed. PMID- 15538027 TI - Youth violence: an exploratory study of a treatment program in a central Illinois county. AB - A local treatment provider has developed an OPTIONS program to prevent adolescent aggression. The program is designed for youths who displayed threatening or abusive behaviors, particularly toward family members. This study described the characteristics of juveniles and their families who were referred to the program and if the program participation helped reduce the severity of violence and improve the communication skills among program participants. Referrals came from law enforcement, juvenile court services, school resource officers, and parents. Information on 100 juveniles and their families were provided by the counselors of the program. The findings showed that a large percentage of juveniles came from unstable families with a family history of criminal convictions, substance abuse, or mental illness. Program participants showed an improvement in communication skills and reduction in their levels of dangerousness. PMID- 15538028 TI - Jail drug and alcohol treatment program reduces recidivism in nonviolent offenders: a longitudinal study of Monroe County, New York's, Jail Treatment Drug and Alcohol Program. AB - Substance abuse treatment has become the new fashion for reducing recidivism among inmates. But the question is, does this work? Various studies have been done tracking the same cohort of inmates over time to assess the validity of treatment. This study assesses one treatment program's success over 5 years to determine if drug and alcohol treatment reduces recidivism among nonviolent, short-term (sentence of less than a year) inmates. Monroe County's drug treatment program demonstrates that for 1 year after receiving the treatment, three different cohorts of nonviolent, short-term inmates (1995, 1998, and 2000) were found to be substantially less likely to be recidivists than control group inmates. PMID- 15538029 TI - Assessing the correctional orientation of corrections officers in South Korea. AB - The correctional goal in South Korea has recently changed from the straightforward punishment of inmates to rehabilitation. Currently, emphases are being placed on education, counseling, and other treatment programs. These changes have consequently begun to also change the corrections officers' roles from a purely custodial role to a human service role, in which officers are expected to manage rehabilitation and treatment programs. Despite these changes, few studies have examined the attitudes of corrections officers toward rehabilitation programming. This is an important dimension to examine in rehabilitation programming, as corrections officers play a major role in the delivery of institutional programs. This study examines the attitudes of South Korean corrections officers toward rehabilitation programs. Approximately 430 corrections officers were sampled. Results show that correctional attitudes are largely influenced by not only officers' own motivations for joining corrections but also by institutional factors such as job stress. Policy implications are discussed. PMID- 15538030 TI - Crime scene staging and its detection. AB - Crime scene staging is a recognized phenomenon within the criminal justice field. This article defines the occurrence for staging, identifies the motives for staging, and identifies the act that precipitated the staging. It reports the results of an exploratory survey of 20 well-versed and highly trained law enforcement officers' experience with staging in fatal and nonfatal incidents, and it reports the most commonly observed forms of crime scene staging. Investigative strategies and indicators of staging are also presented. PMID- 15538031 TI - Commentary: The globalization of knowledge. PMID- 15538034 TI - Measurement of three major subtypes of child neglect. AB - This study examines the relationships among three major subtypes of neglect (physical, psychological, and environmental), a summary measure of neglect, and neglect as defined by Child Protective Services (CPS). The predictive validity of the subtypes of neglect assessed when children were 5 years of age was examined using children's behavior at age 6. Study findings include modest to moderate correlations among the neglect subtypes, CPS neglect, and children's behavior. After controlling for CPS neglect, the subtypes remained predictive of children's behavior. The set of three subtypes was more strongly related to children's behavior, particularly maternal report of internalizing problems, than was the summary neglect measure. These findings suggest that considering the subtypes of neglect may enhance our understanding of neglect and lead to interventions tailored to families' specific needs. PMID- 15538035 TI - Adolescent neglect and alcohol use disorders in two-parent families. AB - Parental responsibilities to adolescents include supervision and emotional support, and variations in these parenting behaviors have been demonstrated to influence adolescent alcohol involvement. This study developed a scale-based method for identifying adolescents with low-parent involvement and examined effects on the development and course of alcohol use disorders (AUDs). The participants were 361 adolescents (ages 14 to 17 years) from two-parent families recruited from clinical and community sources. Cluster analysis of questionnaire items describing mother and father involvement identified 75 adolescents with low parent involvement (i.e., Neglect). Compared with reference adolescents, Neglect adolescents were significantly more likely to be influenced by social pressure to drink alcohol. Among community participants, Neglect adolescents were more likely to develop AUDs. Among adolescents receiving treatment for AUDs, those in the Neglect group showed more improvement during a 1-year follow-up period. The results indicate that inadequate parent involvement may be a form of neglect. PMID- 15538036 TI - Reliability and validity of the mother-child neglect scale. AB - The current study offers a new way to ascertain information about child neglect by evaluating a self-report measure of neglectful behaviors, the Mother-Child Neglect Scale (MCNS). The scale was modified from an existing self-report measure, the Neglect Scale (NS), which was originally designed to measure personal histories of neglect. One hundred adolescent mothers were administered the MCNS and NS in two occasions via phone interviews. In addition, maternal abuse potential and the quality of mother-child interactions were assessed when children were age 3 and 5 years. The MCNS had high internal consistency and moderate test-retest reliability. Convergent validity was found between the MCNS and maternal histories of neglect as well as observed parenting behaviors and child abuse potential; those relationships remained after social desirability was controlled. This research tool may prove useful, in conjunction with existing assessment instruments, in determining the type and severity of past neglectful behaviors. PMID- 15538037 TI - A prospective analysis of the relationship between reported child maltreatment and special education eligibility among poor children. AB - This study presents data from the first large-scale longitudinal study to track the involvement of children reported for maltreatment in both the special education and child welfare systems. A range of state and local administrative databases were combined and cross-sector service histories were established for 7,940 children who had received Aid to Families With Dependent Children between 1993 and 1994. The authors address the following questions: (a) Is maltreatment associated with entry into special education after controlling for other factors? (b) among maltreated children, does maltreatment type or child welfare service use predict special education eligibility? and (c) what is the relationship between maltreatment type and type of educational disability? Results indicate that child maltreatment system involvement generally predates special education entry and is predictive of entry even after controlling for other factors. A range of other associations between factors such as child and maternal characteristics, services received, maltreatment type, and special education classification are detailed. PMID- 15538038 TI - Understanding the risks of child neglect: an exploration of poverty and parenting characteristics. AB - A strong association between poverty and child neglect has been established, but the mechanisms that explain this relationship have not been clearly articulated. This research takes advantage of survey and child maltreatment administrative data about families with young children and assesses the influence of poverty and parenting characteristics on subsequent child neglect. The authors find that indicators of poverty, such as perceived material hardship and infrequent employment, and parenting characteristics, such as low parental warmth, use of physical discipline, and allowing a child to engage in frequent television viewing, are predictive of child neglect. Parenting characteristics do not appear to mediate the link between perceived hardship and neglect, although they suppress the link between employment and neglect. Results from this study provide information that is highly relevant to the approach and design of child maltreatment prevention and intervention strategies. PMID- 15538039 TI - Development and preliminary psychometric properties of the multidimensional neglectful behavior scale-child report. AB - This article describes the development and psychometric properties of the Multidimensional Neglectful Behavior Scale-Child Report (MNBS-CR). The measure is broadly conceptualized to tap child neglect across four core domains: cognitive, emotional, physical and supervisory neglect, and it assesses exposure to violence, alcohol-related neglect, abandonment, and children's appraisals of parenting. Features include pictorial items, audio computer-assisted testing, and programming by age and gender of the child and caregiver. A clinical sample of 144 children, age 6 to 15 years, and a comparison sample of 87 children were tested. Results showed that the MNBS-CR has high reliability, with higher reliability found for older children (alpha = .94) than for younger children (alpha = .66). Among older children, the MNBS-CR Supervisory scale was significantly associated with the Child Behavior Check List (CBCL), and total MNBS-CR scores were significantly associated with clinician reports of behavioral disorders. Younger and older neglected children scored significantly higher on the MNBS-CR than community children. PMID- 15538040 TI - Effects of toremifene (TOR) and tamoxifen (TAM) on serum lipids in postmenopausal patients with breast cancer. AB - This study clarified the difference in the effects on serum lipids between toremifene (TOR) and tamoxifen (TAM). To remove influencing factors, we investigated adjuvant therapy for hormone receptor-positive patients with breast cancer without lymph node metastasis. The subjects were 65 patients who were enrolled in a multicenter randomized comparative study between April 1997 and March 2001. As adjuvant therapy, 20 mg of TAM or 40 mg of TOR was administered for 1 year. The levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), apolipoprotein A-1 (Apo A-1), apolipoprotein A(Apo B), and lipoprotein a (Lp(a)) were measured prior to administration and 3, 6, and 12 months after the start of administration. TC, LDL-C, Lp(a) and Apo B significantly decreased from the third month of administration compared with values before the start of administration in both the TOR and TAM groups. HDL-C significantly increased from the third month only in the TOR group. TG significantly increased in the TAM group but significantly decreased in the TOR group in the 12th month of administration. When these two groups were compared, HDL-C was significantly higher (p < 0.01) and TG was significantly lower (p < 0.01) in the TOR group in the 12th month. Improvement of abnormal values of TG, HDL-C and LDL-C was better in the TOR group than in the TAM group after administration for 12 months. The effect on lipid metabolism showed different profiles between the two selective estrogen receptor modulators (SERMs), and TOR gave better results than TAM. PMID- 15538041 TI - Crossover trial for lipid abnormality in postmenopausal breast cancer patients during selective estrogen receptor modulators (SERMs) administrations. AB - The objective of this study was to evaluate the different profiles of serum lipids resulting from the administration of selective estrogen receptor modulators (SERMs). Postmenopausal primary breast cancer patients (n = 197) with node-negative, hormone receptor-positive who were treated at our department or in other related medical institutions from April 1997 through March 2001 were given adjuvant therapy. The adjuvant therapy included 1 year's administration of tamoxifen (TAM) 20 mg or toremifene (TOR) 40 mg. The profiles of serum lipids such as total cholesterol (TC), high-density lipoprotein cholesterol (HDL) and triglyceride (TG) were observed. After 1 year administration TC had significantly decreased (p < 0.001) both in the TAM group and the TOR group, but no significant difference was found between these groups (p = 0.249). HDL had significantly decreased in the TAM group (p < 0.001), while it had significantly increased in the TOR group (p < 0.001), and a significant difference was found between the groups (p < 0.001). TG had significantly increased in the TAM group (p < 0.001) but significantly decreased in the TOR group (p < 0.001). The medication was switched in those who still had abnormal lipid metabolism and given to them for another year. After 1 year from the crossover TC and HDL had increased to the levels of before administration (p < 0.001) and TG had decreased in those (n = 57) whose medication was switched from TAM to TOR. While TC had decreased and TG had increased in those (n = 23) whose medication was switched from TOR to TAM (p < 0.001). The above findings have suggested that TOR provides better profiles of lipid metabolism than TAM. PMID- 15538042 TI - Expression of FGF-2 alters focal adhesion dynamics in migration-restricted MDA-MB 231 breast cancer cells. AB - Basic fibroblast growth factor (FGF-2) expression takes place during morphogenic differentiation of mammary ducts and is lost in breast cancer. Forced re expression of FGF-2 in breast cancer cell lines induces a more differentiated phenotype and inhibits motility by unknown mechanisms. Here we demonstrate that MDA-MB-231 cells with encumbered motility due to forced re-expression of FGF-2 have activated focal complexes as determined by immunoprecipitation/western blotting and immunofluorescence staining with antibodies to FAK, p130Cas, paxillin, vinculin and phosphotyrosine. The activation of the focal adhesion complexes results in loss of stress fibers associated with malignant transformation of mammary epithelial cells and the formation of circumferentially distributed actin bundles associated with non-transformed mammary epithelial cells. These effects require continuous FGF-2 expression, as the effects of exogenous recombinant FGF-2 are only small and transient. FGF-2 expression results in an increase in integrin alpha 3 expression and decreases in integrin beta 1 and beta 4 expression. These changes, however, induce only a small decrease in adhesion to uncoated and fibronectin-coated tissue culture dishes suggesting that the primary cause of impaired motility is due to intrinsic signaling. These data suggest that FGF-2-inhibits motility in breast cancer cells by stabilization of focal complexes and induction of a more differentiated phenotype with disruption of stress fiber formation and a characteristic cortical actin distribution. PMID- 15538043 TI - Recognition of HLA-A2-restricted mammaglobin-A-derived epitopes by CD8+ cytotoxic T lymphocytes from breast cancer patients. AB - A breast cancer-associated antigen, mammaglobin-A, is specifically expressed in 80% of primary breast tumors. The definition of immune responses against this highly expressed breast cancer-specific antigen should be of great value in the development of new therapeutic strategies for breast cancer. Thus, the purpose of this study was to identify HLA-A2-restricted mammaglobin-A-derived epitopes recognized by CD8+ cytotoxic T lymphocytes (CTL). We identified seven mammaglobin A-derived candidate epitopes that bind the HLA-A2 molecule (Mam-A2.1-7) by means of a HLA class I-peptide binding computer algorithm from the Bioinformatics & Molecular Analysis Section of the National Institutes of Health. Subsequently, we determined that CD8+ CTLs from breast cancer patients reacted to the Mam-A2.1 (83 92, LIYDSSLCDL), Mam-A2.2 (2-10, KLLMVLMLA), Mam-A2.3 (4-12, LMVLMLAAL), Mam-A2.4 (66-74, FLNQTDETL), and Mam-A2.7 (32-40, TINPQVSKT) epitopes using an IFN-gamma ELISPOT assay. Interestingly, healthy individuals also showed high reactivity to the Mam-A2.2 epitope. Two CD8+ CTL lines generated in vitro against TAP-deficient T2 cells loaded with the candidate epitopes showed significant cytotoxic activity against the Mam-A2.1-4 epitopes. These CD8+CTL lines recognized a HLA-A2+breast cancer cell line expressing the Mam-A2.1 epitope. In addition, DNA vaccination of HLA-A2+/human CD8+ double-transgenic mice with a DNA construct encoding the Mam A2.1 epitope and the HLA-A2 molecule induced a significant expansion of epitope specific CD8+ CTLs that recognize the same HLA- A2+/Mam-A2.1+ breast cancer cell line. In conclusion, these results demonstrate the immunotherapeutic potential of mammaglobin-A for the treatment and prevention of breast cancer. PMID- 15538045 TI - Clinicopathological feature and long-term prognosis of apocrine carcinoma of the breast in Japanese women. AB - Because of the rarity of apocrine carcinoma and lack of standardized criteria for the diagnosis, the definitive conclusions of clinicopathologic features and the prognosis has not been determined. We retrospectively examined data on 2091 curatively treated Japanese patients with primary breast carcinoma. Among them, 33 (1.6%) who had been diagnosed of apocrine carcinoma were reviewed. Compared with non-apocrine carcinoma, apocrine carcinoma was characterized by less positive rates of ER and PR, and by frequent rates of unilateral multicentric breast carcinoma with significant difference. The clinicopathological factors influencing 12-year survival rate were lymph node metastasis, lymphatic involvement and vascular involvement. There was no difference in survival rates at 10 years after operation between apocrine carcinoma and non-apocrine carcinoma. Our result shows unique hormone response and unilateral multicentricity are only typical clinicopathological features of apocrine carcinoma. PMID- 15538044 TI - Clinical experience with axillary presentation breast cancer. AB - We present our experience of 50 cases of occult primary tumours presenting as axillary metastases, all with histological report of adenocarcinoma compatible with mammary carcinoma. After bilateral US and mammography, with MRI and mammoscintigraphy where necessary, ipsilateral breast cancer was suspected in 23 cases and quadrantectomy performed. Breast cancer was found only in 12 (24%). In the other 27 women there was no clinical or instrumental suspicion of breast cancer or other primary disease site, so the main treatment was complete axillary dissection plus radiotherapy to the ipsilateral breast (given to all patients). Chemotherapy alone was given to 27 patients, hormone treatment to 5 patients, and both to 18. Mean follow-up is 41.3 months (range 108-1). Thirty-nine (84%) patients are alive with no evidence of disease, two are alive with breast disease, five patients have died of metastatic disease (with no evidence breast disease). Our experience, like that of the literature, confirms that the breast should be extensively investigated but that blanket investigations are not usually revealing. We present guidelines for the work-up of patients presenting with axillary disease. PMID- 15538046 TI - Combined effect of GSTM1, GSTT1, and COMT genotypes in individual breast cancer risk. AB - Our previous studies suggested that both catechol O-methyl transferase (COMT) and glutathione S-transferase (GST) M1 and T1 genotypes are associated with breast cancer risk. Here we extended the studies to evaluate the potential combined effect of these genotypes in individual breast cancer risk. Incident breast cancer cases (n = 202) and controls (n = 299) with no previous cancer were recruited from three teaching hospitals in Seoul in 1996-1999. Information on putative risk factors was collected by interviewed questionnaire. PCR-based methods were used for the genotyping analyses. Odds ratios (ORs) and 95% confidence (CIs) intervals were estimated by unconditional logistic regression after adjustment for known or suspected risk factors of breast cancer. Among pre menopausal women the low activity associated (COMT *L) allele containing genotypes and the GSTM1 null genotype posed increased risks of breast cancer with ORs of 1.7 (95% CI = 1.0 - 2.8) and 1.7 (95% CI = 1.0-2.8), respectively. A marginally significant effect of GSTT1 null genotype was also observed when the total study population was considered (OR = 1.3, 95% CI = 1.0-2.1). When the combined genotype effects were examined, the concurrent lack of GSTM1 and GSTT1 genes posed a more than 2-fold risk of breast cancer (OR = 2.2, 95% CI = 1.2 3.9); this effect was mainly attributable in pre-menopausal women (OR = 3.2, 95% CI = 1.5-7.2). Moreover, the breast cancer risk increased in parallel with the number of COMT , GSTM1 , and GSTT1 at-risk genotypes (p for trend = 0.003). This association was particularly clear in pre-menopausal women among whom combination of all three high-risk genotypes posed a 4.1-fold breast cancer risk (95% CI = 1.4-12.7) compared with pre-menopausal women without at-risk genotypes (p for trend = 0.001). The trend was more pronounced in women with BMI greater than 22 kg/m2 (p for trend < 0.001) and high-risk status of parity factor (nulliparous or women with the first full term pregnancy at age of over 25-year-old) (p for trend = 0.013). These results suggest the combined effect between reproductive factors and GSTM1, GSTT1 and COMT genotypes in human breast carcinogenesis. PMID- 15538047 TI - Factors associated with circulating levels of insulin-like growth factor-I and insulin-like growth factor binding protein-3 in 740 women at risk for breast cancer. AB - Prospective studies have shown an association between elevated plasma levels of insulin-like growth factor-I (IGF-I) and/or decreased levels of its major circulating carrier protein insulin-like growth factor binding protein-3 (IGFBP 3) and increased risk of major cancers. Identifying the factors which affect these biomarkers is of particular interest as subjects at increased risk could benefit from lifestyle changes, and/or chemoprevention intervention. We evaluated the association between constitutional, hormonal and clinical factors and IGF-I and IGFBP-3 in 740 women, including 376 unaffected women and 364 women with intraepithelial neoplasia (IEN) or early invasive breast cancer enrolled in breast cancer chemoprevention trials, conducted at a single institution. Age, body mass index (BMI), height, waist to hip girth ratio (WHR), parity, menopausal status, age at menarche, number of affected first degree relatives, number of biopsies and breast cancer status were considered in the analysis. Women with early breast cancer had 21% higher IGF-I levels (p = 0.033) and 19% higher IGF I/IGFBP-3 molar ratio (p = 0.047) than unaffected women. In unaffected women, age was negatively associated with IGF-I (p = 0.002) and IGF-I/IGFBP-3 (p = 0.001), while age at menarche was negatively associated with IGFBP-3 levels (p = 0.043). In women with IEN or early breast cancer, IGF-I levels were negatively associated with age (p < 0.001), and positively associated with prior biopsies for benign disease (p = 0.013), while age, parity and menopausal status were significant predictors of IGF-I/IGFBP-3 molar ratio. We conclude that circulating IGF-I levels are higher in women with prior breast cancer compared to unaffected women, and that IGF-I and/or IGFBP-3 levels are influenced by age and by reproductive and hormonal factors. These findings support their putative role as breast cancer risk biomarker. PMID- 15538048 TI - Prognostic value of matrix metalloproteinases (MMP-2 and MMP-9) in patients with lymph node-negative breast carcinoma. AB - Twenty-five to thirty percent of patients with node-negative breast cancer are expected to relapse following surgery, therefore great efforts have been made to identify new prognostic markers that could be useful in defining patients for additional therapy. The expression of MMP-2 and MMP-9 has been associated with high potential of metastasis in several human carcinomas including breast cancer. In the present study we examined the prognostic value of immunoreactive MMP-2/MMP 9 protein in 270 consecutive lymph node negative cases who received radical mastectomy or modified radical mastectomy. Among the patients, 211 cases received adjuvant endocrine therapy and/or adjuvant chemotherapy. Using immunohistochemical assay, we found that 56.7% of the resected tumors were positive for MMP-2 whereas 59.6% of the samples were positive for MMP-9. Chi2 test demonstrated a significant direct association between MMP-2 and MMP-9 (p < 0.001); positive immunostaining of MMP-2 was significantly related to higher tumor grade (p < 0.001) and larger tumor size (p = 0.012); positive immunostaining of MMP-9 was significantly related to higher tumor grade (p = 0.002). In univariate analysis, using Cox-proportional hazard model we found MMP 2, MMP-9 and the co-expression of MMPs (MMP2/MMP9) were significantly associated with patients' relapse free survival (p = 0.016, 0.015 and 0.013 respectively) but not overall survival (p = 0.122, 0.320 and 0.091 respectively). Log-rank test also showed that MMP-2, MMP-9 or the co-expression of MMP2/MMP9 was unfavorable prognostic factor for relapse free survival but not overall survival. In subgroup analysis, we found MMPs were more prognostic for patients with no adjuvant treatment than for patients with adjuvant therapy. In multivariate analysis, using Cox-proportional hazard model we found co-expression of MMPs, larger tumor size and higher tumor grade were unfavorable for relapse free survival (p = 0.038, 0.007 and 0.015 for each). We concluded that MMP-2 and MMP-2 are unfavorable prognostic factors in breast cancer patients. They might be potential predictive factor for adjuvant systemic therapy. The co-expression of MMP-2 and MMP-9 has significantly prognostic value in node-negative patients. PMID- 15538049 TI - Flor-Essence herbal tonic does not inhibit mammary tumor development in Sprague Dawley rats. AB - BACKGROUND: Women who are diagnosed with breast cancer often self-administer complementary and alternative medicines to augment their conventional treatments, improve health, or prevent recurrence. Flor-Essence tonic is a complex mixture of herbal extracts used by cancer patients because of anecdotal evidence that it can treat or prevent disease. METHODS: Female Sprague-Dawley rats were given water or exposed to 3 or 6% Flor-Essence beginning at 1 day of age. Mammary tumors were induced with a single oral 40 mg/kg/bw dose of dimethyl-benz[a]anthracene at 50 days of age and sacrificed at 23 weeks. Rats were maintained on AIN-76A diet. RESULTS: Control rats had palpable mammary tumor incidence of 51.0% at 19 weeks of age compared to 65.0 and 59.4% for the 3 and 6% Flor-Essence groups respectively. Overall, no significant difference in time until first palpable tumor was detected among any of the groups. At necropsy, mammary tumor incidence was 82.5% for controls compared to 90.0 and 97.3% for rats consuming 3 and 6% Flor-Essence, respectively. Mean mammary tumor multiplicity (+/-SES) for the controls was 2.8 (+/-0.5) and statistically different from the 3 or 6% Flor Essence groups with 5.2 (+/-0.7), and 4.8 (+/-0.6), respectively (p < or = 0.01). As expected, the majority of isolated tumors were diagnosed as adenocarcinomas. CONCLUSIONS: Flor-Essence can promote mammary tumor development in the Sprague Dawley rat model. This observation is contrary to widely available anecdotal evidence as well as the desire of the consumer that this commercially available herbal tonic will suppress and/or inhibit tumor growth. PMID- 15538050 TI - Frequent activation of CArG binding factor-A expression and binding in N-methyl-N nitrosourea-induced rat mammary carcinomas. AB - We previously identified a positive transcriptional element identical to human Ha ras response element (HRE) within the promoter of the rat Ha-ras gene. We further identified CArG binding factor A (CBF-A), a member of heterogeneous nuclear ribonuclear protein (hnRNP) gene family, as a trans-acting factor that binds the HRE sequence with high affinity in rat mammary carcinoma cells. To determine if activation of CBF-A plays a role in tumor development in vivo , we investigated CBF-A expression and binding activity in rat mammary tumors induced by N-methyl-N nitrosourea. We found that approximately 82% of tumors expressed CBF-A at levels that were 3-20 fold higher than detected in normal mammary gland. Moreover, elevated CBF-A protein levels were invariably associated with increased binding activity to the HRE. CBF-A mRNA levels in tumors were on average elevated only two fold as compared to normal mammary gland, indicating that increased CBF-A protein levels in tumors resulted from both translational and/or post translational regulation. The level of CBF-A expression in mammary tumors was independent of Ha-ras mutational status. Together, these findings indicated that deregulation of CBF-A contributes to mammary carcinogenesis via a mechanism that is distinct from its hnRNP functions in binding and post-transcriptional regulation of RNA. PMID- 15538052 TI - The metabolic syndrome: what to treat, how to treat, what are the goals? AB - Metabolic syndrome is a cluster of risk factors for cardiovascular disease that include obesity, atherogenic dyslipidemia, raised blood pressure, and insulin resistance. The growing trend of obesity is associated with increased prevalence of metabolic syndrome. Optimizing diet and exercise are still the leading therapy for controlling the metabolic syndrome. Based on the current evidence, further emphasis should be placed on aggressive management of other metabolic risk factors such as high blood pressure and dyslipidemia. PMID- 15538053 TI - Intraoperative echocardiography in minimally invasive cardiac surgery and novel cardiovascular surgical techniques. AB - Minimally invasive cardiovascular surgery has been developed in an effort to decrease hospital length of stay and cost by decreasing pulmonary and neurologic complications, pain, chest trauma, and infection. The smaller incisions provide less direct cardiac visualization and exposure, which makes transesophageal echocardiography an essential and integral part of these innovative procedures. Transesophageal echocardiography does not obstruct the surgical field and can perform a full evaluation of the heart. This includes assessment of the aorta and great vessels, valve function, ventricular wall motion, placement of catheters and cannulae, hemodynamic monitoring, and immediate evaluation of surgical results and complications. PMID- 15538054 TI - The diagnostic evaluation of deep vein thrombosis. AB - Due to the morbidity and mortality associated with either untreated disease or inappropriate anticoagulant therapy, accurate diagnosis of venous thromboembolism is essential. As venography, the current gold standard test for deep vein thrombosis (DVT), is both invasive and costly, noninvasive diagnostic strategies for diagnosing DVT have been developed. Noninvasive tests often have to be combined to either raise the post-test probability of disease to a level justifying treatment or lower it to a level at which withholding treatment is warranted. Diagnostic algorithms involving clinical assessment, venous ultrasonography, and D-dimer testing have been validated in management trials of patients with DVT. The optimal strategy at individual institutions is dependent on local expertise and cost. Magnetic resonance venography has the potential to be used as a stand-alone test for DVT but requires further evaluation. PMID- 15538055 TI - Facilitated percutaneous coronary intervention in acute myocardial infarction: attractive concept but difficult to prove! AB - Facilitated percutaneous coronary intervention (PCI) refers to a strategy of immediate PCI following the administration of pharmacological therapies in acute myocardial infarction. It has evolved primarily from the time delays (due to geography or logistics) in getting acute myocardial infarction patients to the catheterization laboratory and the associated irreversible loss of myocardial muscle that occurs as door-to-balloon time increases. Facilitated PCI provides an opportunity to start treating many of these patients before they reach the catheterization laboratory and provides an ability to open the infarct-related artery before PCI, which is associated with better outcomes for AMI patients. Pharmacological strategies before PCI include: thrombolytic therapy, glycoprotein IIb/IIIa inhibitor alone, or a combination of thrombolytic therapy plus glycoprotein IIb/IIIa inhibitor. Initial results of angiographic studies show better patency with the latter strategy but at the expense of higher bleeding event rates. Ongoing trials are evaluating different combinations of thrombolytic and glycoprotein IIb/IIIa inhibitor therapy. PMID- 15538056 TI - Cerebral dysfunction following coronary bypass: fact or fiction? PMID- 15538057 TI - Cerebral dysfunction following cardiopulmonary bypass is a fact: what are the solutions? AB - Focal and diffuse cerebral dysfunction following cardiopulmonary bypass are multifactorial complications that occur with coronary artery bypass grafting. Controversy exists regarding quantification and duration of diffuse cerebral dysfunction. Multiple measures can be undertaken from preoperative screening to intraoperative and postoperative management to decrease the most common complication occurring after coronary artery bypass surgery. PMID- 15538058 TI - Specialty hospitals kill community hospitals: a specious argument. PMID- 15538060 TI - Specialisation in cardiology. PMID- 15538061 TI - A tale of two ventricles: unusual causes of left ventricular hypertrophy. PMID- 15538062 TI - Circumflex coronary occlusion: classic ECG. PMID- 15538063 TI - Hippocrates, Maimonides, and End-of-Life Issues. PMID- 15538064 TI - Effects of a long-term, community-based cardiac rehabilitation program on middle aged and elderly cardiac patients. AB - The purpose of this retrospective study was to compare the effects of a 2-year, community-based cardiac rehabilitation exercise program on cardiovascular fitness, body fatness, and blood lipids in middle-aged (65 years, n=40) male cardiac patients. Estimated maximal metabolic equivalents increased in both groups; however, the increase was greater for middle-aged patients (p=0.003). High-density lipoprotein cholesterol level increased significantly after 1 year in both groups, but the change was greater for the middle-aged subjects by Year 2 (p=0.02). The total cholesterol/high density lipoprotein cholesterol ratio and serum triglyceride levels decreased in both groups, whereas total cholesterol and low-density lipoprotein cholesterol levels decreased only in the elderly group (p<0.01). Body fatness did not change in either group. These findings reinforce the importance of referring elderly as well as middle-aged patients to community-based cardiac rehabilitation exercise programs. PMID- 15538065 TI - Mode of death from congestive heart failure: implications for clinical management. AB - The care of the end-stage patient has not been extensively studied, and little is known about best care practices. Therefore, using new definitions for mode of death due to heart failure, we performed a retrospective chart review of records from a university-based heart failure disease management program to characterize the population of patients dying from heart failure and to define clinical predictors that identify patients who will likely die of metabolic and/or progressive causes. Of 74 deaths recorded over a 60-month period, 17.6% and 21.3% were deemed to be metabolic or progressive, respectively. Utilization of resources was considerable, and only a small number of patients died while in hospice. Patients who required continuous inotropic support and those with preexisting renal failure were at highest risk for non-sudden cardiac death. We conclude that prospective identification of patients at risk for metabolic and progressive heart failure death is possible. The numbers of these patients is likely to increase in an era of implantable cardioverter-defibrillators. Intervention studies designed to evaluate and improve strategies that emphasize symptom control should target this group. PMID- 15538066 TI - Breaking the "bad" news to patients and families: preparing to have the conversation about end-of-life and hospice care. AB - The ability to compassionately communicate difficult or "bad news" to a patient and family is essential to the provision of quality care at the end-of-life. Preparing to have these conversations requires a deliberate multilevel approach that includes: 1) becoming comfortable with end-of-life issues; 2) understanding the scope of the experiences from the patient's and family's perspectives; 3) understanding the full range of choices and options you can offer patients and families regarding their care (including hospice care); 4) developing a "can do" approach toward your involvement with end-of-life care; and 5) learning to effectively share and receive information in a compassionate manner. The following article explores this multilevel approach in an effort to assist physicians and other health care professions in supporting patients and families in their transition from curative care to hospice and palliative care. PMID- 15538067 TI - End-of-life medical treatment of older cardiac patients. AB - The last decade saw breathtaking advances in the science and technology of heart disease. Fewer patients die from an acute myocardial infarction or acute stroke, or have a sudden death. Thanks to modern technology, people live much longer, but more than half still die of heart disease, mostly chronic. Training programs concentrate on mastery of procedures; the discipline of cardiology has become fragmented into noninvasive, nuclear, invasive, interventional, electrophysiology, heart failure, transplantation, and research. Dying persons expect to receive attention to their spiritual and psychosocial, as well as their physical needs. Ordinary people speak of lack of suffering, no unnecessary interventions that postpone the moment of an inevitable dying, not being a burden on others, and having a sense of control. Their assessment of quality may not match that of doctors who speak in terms of science and technology. Review of the limits of modern technology; understanding of existing laws; the near irrelevance of living wills; the role of the physician in the diagnosis of medical futility; the early indications of depression; and, more importantly, better communication with the patient and his/her loved ones are essential components of the practice of medicine. Cardiologists should differentiate between what should be done and what could be done for their patients. PMID- 15538068 TI - Advance care planning. AB - Advance directives allow patients to have some control over decisions even when they are no longer able to make decisions themselves. All states authorize written advance directives, such as the appointment of a health care proxy, but commonly impose procedural requirements. Some states have restricted the use of oral advance directives, although they are frequently used in everyday practice. Advance directives are limited because they are infrequently used, may not be informed, and may conflict with the patient's current best interests. Moreover, surrogates often cannot state patients' preferences accurately. Furthermore, discussions among physicians and patients about advance directives are flawed. Physicians can improve discussions about advance directives by asking the patient who should serve as proxy and by ascertaining the patient's values and general preferences before discussing specific clinical situations. PMID- 15538069 TI - Better late than never: how congestive heart failure patients die. PMID- 15538070 TI - "Tako-tsubo-like left ventricular dysfunction": a clinical entity mimicking acute myocardial infarction with a favorable prognosis. AB - An emotionally-distressed, elderly Caucasian woman presented with chest pain and hypertension. Electrocardiogram showed inferior ST-segment elevation, and an urgent cardiac catheterization was performed. Coronary angiography revealed normal appearing coronary arteries; however, left ventriculography showed extensive left ventricular apical akinesis. The patient had a mild rise in cardiac enzyme levels indicative of myocardial injury. She was discharged after an uncomplicated in-hospital course. One month later, the left ventricular wall motion abnormality had improved. In this report, the authors discuss this compilation of findings known as tako-tsubo-like left ventricular dysfunction. PMID- 15538071 TI - Ethical issues in the management of geriatric cardiac patients. PMID- 15538072 TI - MADIT II-two years later: have the lessons been learned? PMID- 15538073 TI - Images in geriatric cardiology: live three-dimensional transthoracic echocardiographic assessment of right atrial thrombus. PMID- 15538074 TI - Pharmacologic update: treatment of erectile dysfunction in the elderly with phosphodiesterase type 5 inhibitors: cardiovascular implications. PMID- 15538075 TI - Electrocardiology teacher analysis and review: ventriculophasic sinus arrhythmia. PMID- 15538076 TI - On being blue: argyria still exists. PMID- 15538077 TI - Eponyms equal esteem. PMID- 15538078 TI - The changing pattern of skin diseases. PMID- 15538079 TI - Protective effects of oral antioxidants on skin and eye function. AB - BACKGROUND: Antioxidants applied topically and/or taken systemically may play an important role in counteracting the oxidative injury to lipids and proteins on the skin and eye that are provoked by radical oxygen species. OBJECTIVE: The aim of the study was to control photoprotective activity of the skin and eye through the use of oral antioxidants in 50 smoker volunteers (10 cigarettes/d) aged between 25-36 years and affected by dry skin. METHODS: Oxidative stress was assessed by a randomized double-blind study, and the ROS-Meter System (Dermotech, Rome, Italy). Global visual function was determined by the use of different visual surveillance tests. RESULTS: A 40% decrease of oxidative stress in blood serum was verified, with a clear relationship between global visual function and the oral intake of an oxygenated carotenoid (lutein). CONCLUSIONS: Carotenoids used as nutritional supplements seem to play an interesting photoprotective role for both skin and eyes, eventually increasing the activity of topical sunscreens. PMID- 15538080 TI - Rare clinical form in two patients affected by a new variant of endemic pemphigus in northern Colombia. AB - BACKGROUND: Recently, the authors described a new variant of endemic pemphigus foliaceus in rural areas surrounding El Bagre, Colombia, but without association with malignant tumors. METHODS: The authors' 10-year fieldwork provided the opportunity to observe various manifestations of El Bagre endemic pemphigus foliaceus, including the presence of bilateral plaques in pretibial areas. RESULTS: Based on personal experience and literature reviews, the authors have correlated the auto-antibody profile with the appearance of pretibial plaques. CONCLUSION: Since pretibial plaques occur in patients with both fogo selvagem and El Bagre variants of endemic pemphigus foliaceus, as well as in other forms of pemphigoid, these diseases must be considered in the differential diagnosis of patients with clinical, immunologic, and/or epidemiologic risk factors. PMID- 15538081 TI - Hereditary palmoplantar (epidermolytic) keratoderma: illustration through a familial report. AB - Hereditary palmoplantar keratoderma, a well-known clinical entity, is illustrated through a familial report of an unmarried young man who is the product of a consanguineous marriage (paternal and maternal grandmothers were sisters). The lesions were characterized by immense yellow waxy thickening of the skin surrounded by erythematous border (halo) and fissures/cracks associated with extensive scaling of the palms and soles. The lesions were bilateral and symmetrical. These features were supported by orthokeratotic hyperkeratosis hypergranulosis and acanthosis in hematoxylin-eosin stained tissue sections prepared from the soles. Mycelia/spores could not be identified on Periodic acid Schiff (PAS) reaction. An autosomal dominant trait was revealed through family pedigree. An abridged update to recap the current status is highlighted. PMID- 15538083 TI - Cutaneous zygomycosis following attempted radial artery cannulation. AB - A 70-year-old man was seen in a hospital consultation for evaluation of cellulitis of the left arm. The patient had multiple medical problems, including advanced liver disease due to alcohol, diabetes mellitus, congestive heart failure, atrial fibrillation, chronic renal in sufficiency, and hypopituitarism requiring steroid replacement. Most recently, he was admitted to the intensive care unit, where he required intubation and mechanical ventilation support following respiratory failure secondary to pneumonia. At that time, an attempt was also made to place an arterial line in the left radial artery. The patient had multiple areas of ecchymosis on both arms. A large bulla was found on the lateral aspect of the left wrist several days after the attempted arterial line placement. Subsequently, the lesion drained serosanguineous fluid, and, during the next 2 days, it ulcerated with necrosis extending around the wrist and to the elbow. He was started on ampicillin/sulbactam and clindamycin for presumed necrotizing fasciitis. The surgical service performed a very limited debridement,which was partially limited by his coagulopathy from liver disease. The initial tissue culture was positive only for Enterococcus faecium. At the time of the consultation, his temperature was 95' F (35 degrees C), pulse 82 bpm, respirations 16 BPM, and blood pressure 101/56 mmHg. He was awake but not oriented or responsive. His cardiopulmonary exam was unremarkable. Abdominal exam disclosed ascites. His extremities were all grossly edematous with multiple ecchymoses. His left forearm had a circumferential area of ecchymosis and necrosis with macerated margins, sparing only the lateral ulnar epicondyle, and involving deeper structures of subcutaneous fat and muscle(Figures 1-2 showing evolution of the lesion in a period of 1 week). Small tissue clippings were taken from the edge of the lesion and placed on culture plates. By the next morning, the patient's tissue culture grew a mold, later identified as Rhizopus. Amphotericin B was initiated. Surgical intervention (wide debridement with potential conversion to amputation of the left arm) was considered to offer little benefit in view of the patient's multiple and severe comorbidities and his poor prognosis. Amphotericin B was then stopped; the patient died within a week from his multiple medical complications. The family refused an autopsy. PMID- 15538082 TI - Enbrel (etanercept). PMID- 15538084 TI - Kinerase: the science behind the technology. PMID- 15538085 TI - Peptides: the science behind the technology. PMID- 15538086 TI - Microcystic adnexal carcinoma. AB - A 71-year-old woman presented with a firm flesh-colored plaque with overlying whitish discoloration that had been present on her left cheek for at least 15 years (Figure 1). The lesion measured 1.6 x 1.1 cm in diameter. The patient had a history of radiation treatment for acne. What is your diagnosis? What should be the course of management? PMID- 15538087 TI - Myths and misconceptions: not all that glitters under the watch is gold (nickel) dermatitis. PMID- 15538089 TI - Bowenoid papulosis at the site of prior herpes progenitalis. AB - A 35-year-old dentist came to the authors' attention for papular and vegetating lesions that had appeared on his penile shaft over the last 2 months. The lesions differed in their features: pink and vegetating on the left side, brown and papular on the right side (Figure 1). The obvious clinical diagnoses of genital warts on the left and Bowenoid papulosis on the right were confirmed by punch biopsies, which showed epithelial hyperplasia with diffuse cell vacuolization (koilocytes)in the left biopsy and moderate nuclear dysplasia in the right one (Figure 2). Human papillomavirus phenotyping was not performed. Interestingly, the patient reported a clear medical history of herpes progenital is (only one episode) that had involved the right side of his glans and prepuce 4 years ago. An immunomodulating treatment with imiquimod was started in the attempt to cure both the genital warts and Bowenoid papulosis lesions (imiquimod was applied three times a week for 8 weeks). At the end of the treatment, the genital warts had disappeared, whereas many Bowenoid papulosis lesions were still present on the right side of the penis (Figure 3). Treated with liquid nitrogen as well, the Bowenoid papulosis lesions disappeared. PMID- 15538090 TI - Hyperkeratosis of the heels: treatment with salicylic acid in a novel delivery system. AB - A 43-year-old woman presented with dryness and scaling of the lateral and posterior aspects of both heels, which was diagnosed as hyperkeratotic xerosis (Figure 1). Pertinent medical history included dry skin with winter exacerbation and painful hyperkeratosis of the heels present for many years. The patient applied a topical multivesicular cream formulation of 6% salicylic add (Salex, Healthpoint Ltd., Fort Worth, TX) to one foot b.i.d. The physician was blinded as to which foot was treated. After 2 weeks of treatment, it was apparent that the patient was applying the cream to the right foot, as evidenced by reduced dryness, scaling, and hyperkeratosis (Figure2). The patient continued treatment of the same foot for an additional 2 weeks, revealing a dramatic improvement of the right heel,which appeared smooth and soft and devoid of pain. No irritation was associated with treatment; the patient commented that this was the best her heel had been "in years." Subsequently, the patient treated both heels with salicylic acid 60%, multivesicular cream. A second patient, a 25-year-old woman, was treated for ichthyosis vulgaris and hyperkeratosis of both heels. She presented w ith multiple painful fissures and hyperkeratosis of the posterior heels bilaterally (Figure 3). After I week of topical treatment with salicylic add 6%, multivesicular cream applied b.i.d. to the left heel only, there was rapid resolution of both hyperkeratosis and pain (Figure 4). PMID- 15538091 TI - Pimecrolimus-induced tinea incognito. AB - A 6-year-old boy was brought to his primary care provider by his mother, who complained of a pruritic rash near his right eye. The eruption was described as a small, erythematous, slightly scaly plaque at the lateral margin of the right eyelid. The child was in good health and took no medications. The diagnosis of eczema was made; the patient was treated with pimecrolimus cream b.i.d. to the affected area. After 2-3 days of treatment, the itching and erythema completely resolved; however, a rough and scaly plaque persisted. After 1-2 weeks of treatment, the itching gradually returned, and the lesion began to increase in size. Multiple, similar lesions appeared several centimeters from the initially affected area. Pimecrolimus was discontinued; topical nystatin/triamcinolone ointment was prescribed. The eruption continued to spread, and the patient was referred to dermatology for further evaluation. The patient presented to the dermatology clinic with multiple annular, scaly papules and plaques with central clearing. Excoriations and mild inflammation were noted around all affected areas (Figure). A potassium hydroxide examination of the lesions revealed numerous hyphae. The nystatin/triamcinolone ointment was discontinued; oral griseofulvin was prescribed. The eruption improved dramatically after 3 weeks and eventually cleared completely after 5 weeks of treatment. Topical 2% ketoconazole cream was applied b.i.d. for the final 2 weeks of treatment. PMID- 15538092 TI - Case study: fibrosing alopecia in a pattern distribution localized on alopecia androgenetica areas and unaffected scalp. AB - A 54-year-old man with a 24-year history of androgenetic alopecia was referred to the Department of Dermatological Sciences with follicular inflammatory lesions leading to scleroatrophy in the vertex region (Figure 1) of 1-year duration. These lesions appeared a year ago. There was no previous history of this condition. On examination, the patient showed confluent infiltrative follicular lesions on the frontoparietal and occipital scalp (Figure 2). Some lesions evolved into erosions that developed in ivory white scleroatrophy within weeks. These lesions were localized both in and outside of are as affected by alopecia androgenetica and were associated with mild pruritus. Histopathologic examination, performed on an early lesion of the vertex, documented a mild thinning of follicular epithelium associated with an intense lymphohistiocytic perifollicular infiltrate. The damage of the basal cell layer was limited to the follicle, while epidermis was intact. In particular, follicular keratinocytes under the isthmus showed a very intense degeneration exactly where the infiltrate was the most prominent. The damage of the hair sheath was under the isthmus and involved the lower portions of the follicles (including the hair bulbs). The inflammatory infiltrate was exclusively represented by perifollicular lymphohistiocytes. Finally, a connective fibrotic shell with numerous fibroblasts formed a sheath around the atrophic follicle (Figure 3). Results of laboratory investigations (including complete blood cell counts, basal thyroid-stimulating hormone, C-reactive protein, serum ferritin levels, B and C hepatitis markers, antinuclear antibodies, and cultural examinations) were negative.We diagnosed the patient with fibrosing alopecia in a pattern distribution. PMID- 15538093 TI - New-onset diabetes in the hypertension treatment trials: a point of view. PMID- 15538094 TI - Combination angiotensin receptor blocker/hydrochlorothiazide as initial therapy in the treatment of patients with severe hypertension. AB - Patients with severe hypertension are at high risk for cardiovascular events. The authors hypothesized that initial treatment with a combination angiotensin receptor blocker/diuretic agent would be safe and more effective than initial treatment with a single agent for these patients. In this 6-week, double-blind trial, 585 patients were randomized to losartan/hydrochlorothiazide or losartan as monotherapy and titrated as needed at 2-week intervals to reach goal blood pressure (<90 mm Hg). Almost twice as many patients achieved goal at the primary end point of 4 weeks on 50 mg losartan/12.5 mg hydrochlorothiazide vs. the losartan regimen (50-100 mg; p=0.002). Additionally, almost three times as many patients achieved goal blood pressures at 6 weeks (p<0.001). Adverse experiences on losartan/hydrochlorothiazide (43%) were significantly less than with the angiotensin receptor blocker alone (52.6%). This study confirmed the efficacy and tolerability of initial use of a fixed combination of losartan/hydrochlorothiazide vs. losartan without a thiazide. PMID- 15538095 TI - The role of existing and newer calcium channel blockers in the treatment of hypertension. AB - Calcium channel blockers (CCBs), which include both dihydropyridines such as nifedipine and amlodipine and non-dihydropyridines (verapamil and diltiazem), are among the most widely prescribed agents for the management of essential hypertension. Several large outcome risk trials and comprehensive meta-analyses have found that CCBs reduce the cardiovascular morbidity and mortality associated with uncontrolled hypertension, including stroke. CCBs, however, appear less effective than angiotensin-converting enzyme inhibitors and diuretics for preventing heart failure and myocardial infarction. CCBs are among the agents listed as potential first-line therapy, either alone or in combination with other agents in hypertension management guidelines. Furthermore, CCBs are suitable for add-on therapy in combination with diuretics, angiotensin-converting enzyme inhibitors, and angiotensin-II receptor blockers. CCBs may be partially suitable for patients with comorbid Raynaud's syndrome, isolated systolic hypertension (dihydropyridine), or angina pectoris (non-dihydropyridine). The newer inherently long-acting dihydropyridine agents (e.g., lacidipine, lercanidipine), which are not currently available in the United States, appear to have comparable efficacy to older agents of the dihydropyridine class but may have an improved tolerability profile, especially with regard to peripheral edema. PMID- 15538096 TI - The use of aldosterone receptor blockers in the treatment of hypertension. AB - The emerging role of aldosterone in hypertension and cardiovascular diseases has prompted a renewal of interest in therapeutic approaches designed to interfere with the action of this mineralocorticoid hormone. While spironolactone has long been used for this purpose, side effects, largely attributable to the interaction of this agent with non-mineralocorticoid steroid receptors, has reduced the enthusiasm for its use. Eplerenone, a specific aldosterone receptor blocker with a lower incidence of the sex hormone-related side effects than spironolactone, has been used in several recent clinical trials in hypertension and congestive heart failure. This review will highlight the major findings from these studies. PMID- 15538097 TI - Hypertension curriculum review: epidemiology and the prevention of hypertension. PMID- 15538098 TI - Interview with Jerry Stamler, MD. Interview by Marvin Moser. PMID- 15538099 TI - Reflections in hypertension: heart failure and hypertension: the diastolic dilemma. PMID- 15538100 TI - Case studies in hypertension: presentation with vaginal bleeding and hypertension. PMID- 15538103 TI - Current concepts of pharmacotherapy in hypertension: thiazide-type diuretics: ongoing considerations on mechanism of action. AB - Thiazide-type diuretics have enjoyed a considerable success in the management of hypertension. These drugs have assumed a standard-of-care position in the minds of many health care providers; however, a number of questions remain unresolved in relation to their use. Such questions include issues of mechanism of action, comparability to loop-diuretics in their actions, class-effect, and the basis for their additivity with non-diuretic antihypertensive medication classes. Understanding these issues is important to the effective use of these compounds. PMID- 15538104 TI - Microalbuminuria: definition, detection, and clinical significance. AB - Proteinuria is a sign of abnormal excretion of protein by the kidney but is a nonspecific term including any or all proteins excreted. In contrast, albuminuria specifically refers to an abnormal excretion rate of albumin. Microalbuminuria refers to an abnormally increased excretion rate of albumin in the urine in the range of 30-299 mg/g creatinine. It is a marker of endothelial dysfunction and increased risk for cardiovascular morbidity and mortality especially, but not exclusively, in high-risk populations such as diabetics and hypertensives. Testing for microalbuminuria is now made easy by in-office dipstick tests (semiquantitative) and widely available laboratory testing (quantitative). Physicians should screen all diabetics for albuminuria and strongly consider screening hypertensives to identify those at higher risk for cardiovascular disease. Appropriate intervention, including use of drugs that block the renin angiotensin-aldosterone system, may be appropriate in such cases as suggested by the American Diabetes Association and the Seventh Report of Joint National Committee on the Prevention, Detection, Evaluation, and Treatment of High Blood Pressure. PMID- 15538105 TI - Comparison of commonly used assays for the detection of microalbuminuria. AB - There are a variety of methods for assessing urinary albumin excretion, extending from the very low-range microalbuminuria to higher ranges extending into macroalbuminuria or proteinuria. The recommendation for the initial screening of a new patient is to use a urine dipstick to assess for microalbuminuria. If positive, a spot urine for albumin:creatinine should be measured and reassessed annually. All patients with kidney disease, diabetes, or hypertension and metabolic syndrome should be screened for albuminuria. New methodologies using high-performance liquid chromatography are much more sensitive and specific when compared with older methods of detection and may prove very useful for earlier identification of high-risk patients. This is important since studies have shown that albuminuria levels below the microalbuminuria range, determined by conventional methodologies in uncomplicated essential hypertensive men, are associated with an adverse cardiovascular and metabolic risk profile. High performance liquid chromatography methodology, in contrast to older studies, detects all intact albumin and enables clinicians to assess disease severity and monitor therapeutic effectiveness with confidence in the accuracy of the microalbuminuria data reported to them. PMID- 15538106 TI - Cardiovascular implications of albuminuria. AB - Microalbuminuria is a major independent risk factor for cardiovascular disease (CVD) events in persons with diabetes or hypertension, the general population, and persons with known CVD. Although microalbuminuria is a stronger risk factor in men, women with increased albuminuria levels are also at a higher risk of CVD. Microalbuminuria is an indicator of generalized endothelial injury, a hallmark of systemic atherosclerosis. Treatments that decrease albuminuria, particularly agents that inhibit the renin-angiotensin system, reduce CVD risk in various populations, including those with and without diabetes or hypertension. Whether albuminuria should be a treatment target for CVD is not yet proven. Nevertheless, the measurement of albuminuria is clinically useful to identify high-risk individuals who should receive intensive risk factor management based on current treatment guidelines. PMID- 15538107 TI - Implications of albuminuria on kidney disease progression. AB - Albuminuria is recognized in all hypertension guideline statements as a cardiovascular risk factor and indicator of kidney disease. Recent data also demonstrate a strong association between the presence of microalbuminuria and elevations in C-reactive protein. Thus, the increased membrane permeability that generates microalbuminuria may be secondary to an inflammatory process. Progression from microalbuminuria (>30 and < or =300 mg albumin/g creatinine) to macroalbuminuria (>300 mg albumin/g creatinine) indicates a worsening of vascular disease and the presence of kidney disease. Recent outcome trials of kidney disease progression have demonstrated the best results among those with reductions in albuminuria in concert with blood pressure (BP) reduction. Thus, use antihypertensive agents that not only lower BP but also lower or normalize albuminuria levels. All recent guideline statements support the use of agents that block the renin-angiotensin-aldosterone system as part of a regimen to achieve the BP goal. Further lowering of albuminuria may be achieved by adding either a nondihydropyridine calcium antagonist such as verapamil or diltiazem, or aldosterone receptor blockers. Use of an angiotensin receptor blocker added to an angiotensin-converting enzyme inhibitor or vice versa can further lower albuminuria by an additional 30%-40%, which is not true of the additional lowering of BP. PMID- 15538108 TI - Dietary salt, blood pressure, and microalbuminuria. AB - The relationship between dietary salt, blood pressure, and risk for cardiovascular disease has been debated for decades. Microalbuminuria is a biomarker for both cardiovascular and kidney disease. The presence of microalbuminuria correlates directly with the risk for myocardial infarction and stroke and indicates individuals at risk for the development of progressive kidney disease. Since patients with the metabolic syndrome, diabetes, or chronic kidney disease often are blood pressure salt sensitive, and it is well known that increasing dietary salt may offset both the antihypertensive and antiproteinuric effects of renin-angiotensin system blocking drugs, physicians must consider increased salt intake as a potential modifiable risk factor for progression of chronic kidney disease and possibly even cardiovascular disease. PMID- 15538109 TI - Paraffin section interphase fluorescence in situ hybridization in the diagnosis and classification of non-hodgkin lymphomas. AB - Cytogenetic data can contribute valuable information that may assist in the diagnosis and classification of non-Hodgkin lymphomas and may in some cases also provide prognostic information. Interphase fluorescence in situ hybridization (FISH) studies offer the ability to assess for characteristic cytogenetic abnormalities even when material for standard metaphase cytogenetic analysis is not available. This review discusses the use of FISH in paraffin-embedded material with particular attention paid to the use of intact thin paraffin sections. The basic principles of FISH analysis are summarized, the advantages and disadvantages of analysis of thin paraffin sections rather than intact nuclei are discussed, and the more commonly encountered artifacts are considered. Each of the well-characterized cytogenetic abnormalities that are associated with particular types of non-Hodgkin lymphoma and can be detected with commercially available FISH probes is discussed individually. In particular, their incidence in various types of lymphoma is reviewed, the types of commercially available FISH probes to detect such abnormalities are discussed, and clinical situations where such analysis can be of diagnostic utility are described. PMID- 15538110 TI - Expression microdissection: operator-independent retrieval of cells for molecular profiling. AB - Tissue microdissection is an important method for the study of disease states. However, it is difficult to perform high-throughput molecular analysis with current techniques. We describe here a prototype version of a novel technique (expression microdissection) that allows for the procurement of desired cells via molecular targeting. Expression microdissection (xMD) offers significant advantages over available methods, including an increase in dissection speed of several orders of magnitude. xMD may become a valuable tool for investigators studying cancer or other disease states in patient specimens and animal models. PMID- 15538111 TI - Feasibility of using tissue microarrays for the assessment of HER-2 gene amplification by fluorescence in situ hybridization in breast carcinoma. AB - Tissue microarrays (TMAs) have been commonly used to study protein expression by immunohistochemistry (IHC). However, limited data exist on the validity of using TMAs to study gene amplification. In this study, we evaluated the feasibility of using breast carcinoma TMAs to study HER-2 gene amplification by fluorescence in situ hybridization (FISH). In addition, hormonal receptor status (ER and PR) and HER-2 protein overexpression by IHC were also studied, and results were compared with whole tissue sections. FISH for HER-2 was performed on formalin-fixed paraffin-embedded tissue from 114 invasive breast carcinomas both on whole tissue sections and on TMAs containing the same tumors. The TMA was created using 0.6-mm tissue cores with four sampled cores per tumor from the same tissue block used for whole section FISH. The PathVysion HER-2 probe kit was used for the FISH analysis. A ratio of HER-2:Chromosome17 > or =2.0 was interpreted as positive for gene amplification. The ER or PR was interpreted as positive when nuclear staining was detected in more than 10% of tumor cells. The HER-2 IHC (HercepTest; DAKO Corp, Carpinteria, CA) results were interpreted as 0, 1+, 2+, and 3+ according to standard criteria. The FISH results in the TMA and whole sections were concordant in 99 out of 101 successfully analyzed cases (99%). The FISH scores were consistent among the two to four cores in the majority of the cases. ER and PR results were concordant between whole sections and TMA cores in 97% (107/110) and 89% (97/109) cases, respectively. The overall concordance for HER-2 status by IHC between whole sections and TMA cores was 86% (94 out of 109 cases). TMAs are a reliable approach to study HER-2 gene amplification in a high throughput manner. PMID- 15538112 TI - Genetic and immunophenotype analyses of TP53 in bladder cancer: TP53 alterations are associated with tumor progression. AB - Altered p53 status is a frequent event in bladder cancer and reported to have prognostic significance. We studied the TP53 gene and its product in 76 patients affected with urinary bladder carcinomas by immunohistochemistry (mAb DO-7), polymerase chain reaction single-strand conformational polymorphism (exons 4-8) followed by direct sequencing of shifted bands, and loss of heterozygosity in 17p (p53CA). H-RAS mutations were also studied. The receiver operating characteristic curve and the logistic-regression analysis were used to evaluate the validity of immunohistochemistry in predicting TP53 mutations. A p53-positive nuclear phenotype was defined by a cutoff of 20% tumor cells being immunoreactive and was found in 23 cases, while TP53 mutations were detected in 22 cases, four of them with a negative p53 phenotype. TP53 deletions were identified in 23 cases. No H RAS gene mutations were observed. There was a significant association between phenotype and genotype results. Moreover, a significant association was observed between p53 status and tumor stage and grade, being alterations more common in high-stage and high-grade tumors (both chi2 test; P < .01). Deletion of 17p significantly correlated with tumor stage (P < .01) and grade (P = .01), allelic losses being more common in advanced disease. Data from these studies suggest that genetic assays are necessary for the optimal determination of TP53 alterations, mainly in tumors with a p53 negative phenotype, and especially in early stage tumors for which p53 status may assist in determining its progression to invasive disease. Since p53 alterations are significantly associated to clinicopathological features of poor prognosis, the inclusion of both p53 phenotype and TP53 mutation status into a predictive panel of tumor markers for bladder cancer is recommended. PMID- 15538113 TI - The influence of immunohistochemistry on mRNA recovery from microdissected frozen and formalin-fixed, paraffin-embedded sections. AB - Laser-assisted microdissection (LAM) is now widely used to obtain specific cell populations from heterogeneous tissues. A major disadvantage of LAM is poor tissue morphology during microscopy, in part because coverslips are not used. Immunohistochemical labeling can improve identification of target cells but may affect the subsequent analysis of the microdissected tissue. We studied the effect of immunohistochemistry (IHC) on mRNA recovery from labeled cells after microdissection from both frozen and formalin-fixed, paraffin-embedded (FFPE) sections, using Melan-A and Ki-67 staining in lymph nodes with metastatic melanoma as a model. We developed rapid protocols for immunostaining in an attempt to limit loss of mRNA during procedures. A sensitive real-time quantitative reverse transcription-PCR was used to measure mRNA. We found a marked decrease in the mRNA yield from 500 microdissected cells from frozen and paraffin sections after immunostaining for both markers. Recovery of mRNA decreased by up to 89%, comparing the immunostained with the routinely stained sections. Interestingly, the ratio between mRNA for the two markers was similar in all stains, indicating that immunostained sections may be used for mRNA analysis. We also investigated the effect of storing membrane-mounted sections for microdissection under different conditions. Slides mounted with paraffin sections could be stored at room temperature for up to 90 days with no significant decrease in mRNA recovery. PMID- 15538114 TI - Effect of formalin, acetone, and RNAlater fixatives on tissue preservation and different size amplicons by real-time PCR from paraffin-embedded tissue. AB - RNA recovered from paraffin-embedded tissue has been reported to be a suitable substrate for polymerase chain reaction. During tissue fixation and paraffin embedding, RNA undergoes degradation, but with certain restrictions, it can be used for gene expression studies. At the same time, formalin-fixed, paraffin embedded histopathology archives contain an unestimable collection, which could be analyzed to investigate changes in mRNA expression in pathologic processes. To decide for future tissue conservation of pathology samples, it would be reasonable to satisfy both histologic and molecular biologic needs. The effect of three different fixation methods, RNAlater (SIGMA R 0901, St Louis, MO), acetone, and formalin, were compared by histology, immunohistochemistry, and real-time PCR. To assess tissue structure preservation and antigenicity, hematoxylin-eosin staining and immunohistochemistry were performed; to assess RNA quality, RNA was extracted and the transcription of different amplicon sizes (121, 225, 406 bp for GAPDH; 166, 310, 536 bp for beta globin) were examined on human endometrium samples. The most adequate tissue preservation was found in case of formalin fixation, while there were no significant differences in the three fixatives' yields for various size real-time PCR amplicons. Longer amplicons (above approximately 225 bp) have limited use for gene expression studies, while shorter amplicons could give more reliable results. PMID- 15538115 TI - Overexpression of urinary plasminogen activator (uPA) protein and mRNA in thyroid carcinogenesis. AB - OBJECTIVE: Urinary plasminogen activator (uPA), a protease, is one of the critical components of tumor invasion and metastasis. Its expression in thyroid carcinoma and potential role in thyroid tumorigenesis are unknown. The objective of this study was to determine whether uPA is differentially expressed in benign and malignant thyroid tumors. DESIGN: uPA expression was evaluated by immunohistochemistry (IHC) in 20 thyroid tumors (six classic papillary thyroid cancers (PTC) and three tall cell variants (TCV) and 11 adenomas). To validate IHC results, in situ hybridization was performed on both adenomas and cancer tissues to explore the expression of uPA at the mRNA level. The Fisher exact test was used to compare protein as well as mRNA expressions in adenomas and thyroid cancer. RESULTS: Intense granular cytoplasmic staining for uPA was observed in five of nine (56%) of thyroid cancers: 2/6 classic PTC (33%) and all tall cell variant PTC. Furthermore, uPA mRNA expression was found in the malignant thyroid epithelium but not in adjacent normal thyroid follicles or the stromal elements. None of the adenomas expressed uPA (P = .008). uPA staining was absent in histologically normal follicles adjacent to malignant thyroid follicles. CONCLUSIONS: uPA is expressed in thyroid carcinoma but not in benign adenomas or normal adjacent follicles. The selective expression of uPA in thyroid carcinoma provides evidence that uPA is useful in distinguishing benign and malignant thyroid neoplasms. More importantly, uPA may represent molecular target for therapeutic treatment of this malignancy. PMID- 15538116 TI - Development of 5' nuclease real-time PCR assays for the rapid identification of the burkholderia mallei//burkholderia pseudomallei complex. AB - Burkholderia pseudomallei is the causative agent of melioidosis and was classified as a biologic agent by the Centers for Disease Control and Prevention (Atlanta, GA). Acute melioidosis has a case fatality rate of >40%, and septicemia is fatal in up to 90%. The aim of the study was to design 5'-nuclease real-time PCR assays for the rapid and reliable identification of the B. mallei/B. pseudomallei complex. Real-time PCR assays using TaqMan probes targeting the 16S rDNA and fliC were developed on an ABI Prism 7000 sequence detection system (Applied Biosystems, Foster City, CA). Specificity was assessed with 64 B. pseudomallei, nine B. mallei, 126 other Burkholderia strains of 29 species, and 45 clinically relevant non-Burkholderia organisms. Sensitivity, specificity, and positive and negative predictive value of the assays were 100%. Discrimination between B. pseudomallei and B. mallei, an organism which can be regarded as a clone of B. pseudomallei, could not be achieved. A probit analysis revealed that 7.5 and 52 genome equivalents (GE) of B. pseudomallei could be detected using the fliC and the 16S rDNA assays (P = .05), respectively. In spiked blood samples, the detection limit was approximately 300 and 3.000 GE for fliC and the 16S rDNA, respectively. In conclusion, we recommend the simultaneous use of the 16S rDNA and fliC real-time PCR assays for the rapid and specific identification of the B. mallei/B. pseudomallei complex in positive blood cultures or from suspicious bacterial colonies allowing the early onset of appropriate antibiotic therapy. PMID- 15538117 TI - Brain cytokines and neuropsychiatric disorders. PMID- 15538118 TI - Quality of life outcomes of risperidone, olanzapine, and typical antipsychotics among schizophrenia patients treated in routine clinical practice: a naturalistic comparative study. AB - Findings in previous studies investigating the beneficial effect of risperidone and olanzapine versus typical antipsychotics on quality of life (QOL) are controversial since they did not adjust for various factors contributing to QOL. To test this assumption in a naturalistic cross-sectional design, we evaluated general and domain-specific QOL scores for baseline data of schizophrenia outpatients stabilized on atypical (N = 78, risperidone or olanzapine) and typical (N = 55) agents. Self-report and observer-rated QOL outcomes of both risperidone and olanzapine with typical antipsychotic therapy were compared across demographic, illness-related, and treatment-related factors using analysis of variance, multivariate analysis of variance, and correlation analysis. No significant differences were found in QOL outcomes of risperidone-treated and olanzapine-treated patients. Both self-report and rater-observed QOL measures indicated superiority of atypical over typical antipsychotic agents after adjusting for daily doses, duration of treatment, subjective tolerability, and adjuvant antidepressants. Lower daily doses and longer antipsychotic treatment were associated with better QOL. Self-report and observer-rated QOL scores correlated positively (r = 0.64, P < 0.001). Gender, marital status, age, education, living arrangement and employment status, age of onset, illness duration, symptom severity, emotional distress, subtypes of schizophrenia, and side effects did not affect QOL outcomes in either group. Risperidone and olanzapine revealed an advantage over typical agents in terms of QOL. Findings suggest that when calculating the beneficial effects of atypical antipsychotic therapy on QOL outcomes, daily doses, duration of treatment, and subjective tolerability may be intervening variables and should be adjusted accordingly to clearly appreciate benefits of atypical antipsychotics. PMID- 15538119 TI - Ethnicity and the course of tardive dyskinesia in outpatients presenting to the motor disorders clinic at the Maryland psychiatric research center. AB - BACKGROUND: Although newly emergent tardive dyskinesia (TD) is less of a concern, about one-fourth to one-third of patients on or previously on chronic first generation antipsychotic agents have TD. The long-term course and outcome, as well as their predictors, are unknown. Earlier studies identify ethnicity as one of the risk factors for the development of TD, and case reports have noted a preponderance of African-American males in cohorts of patients with tardive dystonia. The current study examines the anatomic distribution and course of TD in a cohort of schizophrenia patients of European and African descent with TD who were referred to the Motor Disorders Clinic (MDC). METHODS: We evaluated data collected on 1149 TD patients who were given a focused neurologic examination for movement disorders. Movements were evaluated with the MPRC Scale for Involuntary Movements (IMS). All patients met RDC-TD criteria for diagnosis of persistent TD. One to 10-year follow-up data on 528 patients were evaluated to examine the course of TD following recommendations made to referring primary clinicians. Suggested interventions to referring primary clinicians included dose reduction of first-generation antipsychotic medication, or switching to a second-generation antipsychotic. RESULTS: Initial evaluation included 701 European American (EA) patients and 448 African-American (AA) patients. AA patients had a significantly higher proportion of males [chi(1) = 7.50, P < 0.05]. EA subjects had a higher mean age than AA patients 42.8 +/- 11.2 and 39.8 +/- 10.4, respectively [F(1,1147) = 22.27, P < 0.05]. Mean neuroleptic exposure (chlorpromazine equivalents) was similar in both groups after controlling for differences in age.Follow-up data analyzed in 528 patients (329 EA and 199AA) showed a significant ethnicity by TD interaction [F(1,504) = 4.26, P < 0.05]. Examination of body distribution of dyskinetic movements showed an effect of ethnicity. Subsequent analyses suggest EA patients experienced more improvement in TD over the course of follow up [F(1,319) = 22.39, P < 0.05] compared with AAs [F(1,189) = 1.58, P > 0.05]. These findings were unchanged when age, change in antipsychotic drug dose, and duration of follow-up were covaried. CONCLUSION: Reports from earlier studies note ethnicity (African descent) as a risk factor in the development of TD. Our study findings suggest ethnicity might be an important factor in predicting a poor course of TD. PMID- 15538121 TI - Predictors of response to treatment of acute bipolar manic episodes with divalproex sodium or placebo in 2 randomized, controlled, parallel-group trials. AB - Identification and control of factors that are associated with placebo and drug response in acute mania may influence the design and analysis of future trials in this condition. We analyzed pooled data from 2 multicenter, placebo-controlled, randomized trials of divalproex sodium for the treatment of acute manic episodes, using linear and logistic regression to search for common and treatment-specific predictors of response. In both treatment groups, patients with greater numbers of prior hospitalizations, more severe baseline symptoms, or younger age at onset of illness were found to be significantly less likely to respond. The effect sizes associated with these factors were of comparable magnitude to the effect size associated with divalproex treatment. Stratification on the basis of these prognostic factors may improve the power of future trials for the treatment of acute mania and may help explain patient response to treatment. PMID- 15538120 TI - Quetiapine versus placebo in combination with lithium or divalproex for the treatment of bipolar mania. AB - Quetiapine (QTP) combined with lithium (Li) or divalproex (DVP) for the treatment of mania was evaluated in 2 double-blind, placebo-controlled studies. Patients were randomized to 3 or 6 weeks of treatment with QTP plus Li/DVP or placebo (PBO) plus Li/DVP. Quetiapine was dosed up to 800 mg/d; Li was dosed to achieve serum concentrations of 0.7-1.0 mEq/L and DVP to 50-100 microg/mL. A total of 402 patients were randomized: 197 to QTP + Li/DVP and 205 to PBO + Li/DVP. The mean quetiapine dose in responders was 492 (+/-204) mg/d. Improvement in the Young Mania Rating Scale (YMRS) at day 21 in the QTP + Li/DVP group (-15.29) was statistically superior to the PBO + Li/DVP group (-12.19) (P < 0.05). A statistically significant difference in favor of quetiapine was observed within the first week (P < 0.05). Significantly more QTP + Li/DVP patients achieved a response (> or =50% decrease in the YMRS) at day 21 (QTP + Li/DVP, 55.7%; PBO + Li/DVP, 41.6%;P < 0.01). Improvements in Clinical Global Impression-Bipolar Severity of Illness scores by day 21 were also significantly greater in QTP + Li/DVP-treated patients (-1.59) versus PBO + Li/DVP (-1.19) (P < 0.01). Common adverse events (> or =5% and twice that in the PBO + Li/DVP group) in the QTP + Li/DVP group were somnolence, dry mouth, and asthenia. Quetiapine combined with Li/DVP was not associated with extrapyramidal symptoms (including akathisia) or emergent depression. More QTP + Li/DVP-treated patients completed the trial, and there was no difference in discontinuation rates due to adverse events between the two groups. Quetiapine, in combination with lithium or divalproex, is well tolerated and has superior efficacy to lithium or divalproex alone in the treatment of bipolar mania. PMID- 15538122 TI - Impact of myeloperoxidase and NADPH-oxidase polymorphisms in drug-induced agranulocytosis. AB - Therapy of the atypical antipsychotic drug clozapine is limited by a comparatively high incidence of agranulocytosis in 0.8% of patients. This severe side effect is possibly based on the clozapine-mediated stimulation of cytokines and soluble cytokine receptors release, followed by induction of granulocyte proliferation and induction of myeloperoxidase (MPO) and NADPH-oxidase. Because NADPH-oxidase/MPO may oxidize clozapine to highly reactive nitrenium ions, we investigated the role of hereditary polymorphisms in the NADPH oxidase/myeloperoxidase system in agranulocytosis patients who received clozapine (n = 49), ticlopidine (n = 11), and other drugs prior to the event. The low active MPO -436A allelic variant frequency was 22.2% in cases and 19.9% in controls, but AA carriers were overrepresented among cases compared with the sum of AG and GG-carriers (odds ratio 4.16, 95% confidence limits 0.86-20.3, P = 0.056). Particularly in clozapine-induced agranulocytosis, this finding was most pronounced (P = 0.04). In the CYBA gene, encoding the p22phox subunit of the NADPH-oxidase, 2 polymorphisms were investigated. C242T (His72Tyr) had an allele frequency of 31.9% and 32.2% (P = NS) and A640G in the 3'-UTR was less frequent in cases (48.7%) than controls (60.0%), odds ratio 0.63 (0.39-1.02), P = 0.048. CYBA 640GG-carriers were marginally less frequent in cases compared with controls (28.2% vs. 38.7%, P = 0.062). Sequencing the entire coding region of the NADPH subunit CYBB (gpS1phase) disclosed that CYBB is a highly conserved gene, which does not represent a risk factor for clozapine-induced agranulocytosis. The impact of the polymorphic myeloperoxidase, however, needs further verification to predict a patient's risk to develop drug-induced agranulocytosis. PMID- 15538123 TI - Multidimensional sensory phenomena in antipsychotic-induced akathisia. AB - The primary distinguishing features of akathisia in comparison with other extrapyramidal syndromes are the prominent subjective manifestations, which include various sensory symptoms. The sensory symptoms are multidimensional in nature and encompass various forms of bodily and mental sensations. The purpose of the present study was to elucidate the multidimensional aspects of the sensory phenomena associated with antipsychotic-induced akathisia. Seventy stable and chronic schizophrenic subjects receiving maintenance antipsychotic treatment were evaluated for akathisia and other extrapyramidal side effects. Subjective sensory phenomena were evaluated in 3 dimensions (ie, bodily sensations, mental sensations, and autonomic sensations). The frequency of each dimension of these sensory phenomena was compared between the groups with and without akathisia using chi test with Bonferroni correction. The akathisia group (n = 29) reported significantly more frequent focal or generalized bodily sensations than the non akathisia group (n = 41) (P < 0.001). The akathisia group also showed significantly more frequent mental sensations such as mental urge and a feeling of inner tension/pressure (P < 0.001). In autonomic phenomena, there was a trend for the akathisia group to show more frequent autonomic sensations. The most common autonomic phenomena associated with akathisia were palpitation and difficulty breathing. The results of the present study suggest that the sensory phenomena of antipsychotic-induced akathisia are characterized by multidimensional features such as bodily sensations, mental sensations, and several autonomic sensations. The assessment of these multidimensional sensory phenomena would be useful in evaluating the important phenotypic features of akathisia. PMID- 15538124 TI - Obsessive-compulsive disorder in the postpartum: open-label trial of quetiapine augmentation. AB - OBJECTIVE: Postpartum nonpsychotic conditions are routinely treated with antidepressant therapy. However, a subset of this population with comorbid obsessive-compulsive disorder (OCD) is treatment-resistant. Optimal response is obtained by augmentation therapy with novel antipsychotics. The objective of this open-label study was to evaluate clinical response to quetiapine augmentation of SSRIs or SNRIs in treatment-resistant OCD in the postpartum. METHODS: Twenty-two postpartum women diagnosed with OCD as per DSM-IV criteria, who did not respond to at least 8 weeks of SSRI or SNRI monotherapy, were offered a trial of quetiapine augmentation for 12 weeks. Response (defined as >50% reduction in scores) was assessed using the Yale Brown Obsessive-Compulsive Scale (YBOCS) and Clinical Global Impressions scale (CGI). RESULTS: Seventeen patients agreed to a trial of quetiapine augmentation. Three withdrew early due to side effects, and 14 completed the 12-week trial. Of these, 11 responded to treatment within 12 weeks, with a mean (SD) response time of 5.9 (2.6) weeks. The mean (SD) baseline YBOCS score of 24.7 (6.8) dropped to a mean of 10.3 (9.0), with a mean reduction of 59.6%. Mean CGI scores at outcome were 1.9 (1.2). The average dose of response was 112.5 mg (76.4 mg). Sedation was the most commonly reported side effect. CONCLUSIONS: Although limited by lack of controls, this is the first study in a postpartum population where the addition of quetiapine to antidepressant therapy has been shown to be effective for treatment-refractory OCD. Quetiapine deserves further controlled study in this context. PMID- 15538125 TI - The advantages of choosing antiobsessive therapy according to decision-making functioning. AB - OBJECTIVE: Previous studies stressed the role of decision-making functioning in predicting antiobsessive treatment outcome with serotonin reuptake inhibitors drugs in patients with obsessive-compulsive disorder. Nevertheless, the use of an augmentation strategy with atypical antipsychotic drugs has proved to be effective in obsessive-compulsive patients nonresponding to serotonin reuptake inhibitors treatment. We investigated whether the performance at the Iowa Gambling Task (IGT), a used neuropsychologic task which assesses decision-making, can be an effective criterion for pharmacologic treatment choice in these patients and whether the use of different treatment strategies, according IGT performance, can increase the rate of antiobsessive outcome. METHOD: Thirty patients with obsessive-compulsive disorder were treated in a single-blind design with fluvoxamine plus placebo or fluvoxamine plus risperidone according to their IGT performance. Treatment outcome was recorded after 6 and 12 weeks. RESULTS: Patients with good IGT performance showed a good antiobsessive treatment outcome with fluvoxamine only, while only adopting an augmentation strategy with risperidone, the number of responders patients within the subjects with bad IGT performance increased. CONCLUSIONS: IGT performance may be considered an effective criterion for pharmacologic treatment choice in obsessive-compulsive patients given that antiobsessive treatment outcome is increased to 85% of responders choosing an appropriate drug strategy according to the IGT performance. PMID- 15538126 TI - Association between SSRI use and hip fractures and the effect of residual confounding bias in claims database studies. AB - BACKGROUND: Although use of selective serotonin reuptake inhibitor (SSRI) antidepressants has been associated with hip fractures in claims data studies, it has been suggested that such results may be confounded by cognitive and functional status for which no information is available in claims data. Using survey data, we determined the magnitude of such bias and corrected the association between SSRI use and hip fractures accordingly. METHODS: We used the Medicare Current Beneficiary Survey to determine the association between SSRI use and 5 potential confounding factors not measured in Medicare claims data: body mass index, smoking, activities of daily living score, cognitive impairment, and Rosow-Breslau physical impairment scale. For 7126 participants aged > or =65 years, we estimated the association between SSRI use and these potential confounders. Combined with literature estimates of the associations between confounders and hip fractures, we were able to compute the extent of residual confounding bias caused by a failure to adjust for these factors. RESULTS: Comparing SSRI users with nonusers, there was considerable overestimation of an association with hip fractures if activities of daily living scores (+21.5% bias) or Rosow-Breslau impairment scales (+10.6%) are unmeasured in claims data. All 5 unmeasured confounders together resulted in net confounding of +9.6% (range: 0.3% to + 39%). After correction for this bias, the strength of association observed in claims data after bias correction (RR = 1.8; 95% CI = 1.5 to 2.1) was comparable to a recent clinical study (RR = 1.5; 0.6 to 3.8), but the claims data study achieved formal statistical significance due to its much large size (8239 vs. 288 hip fractures). CONCLUSIONS: Epidemiologic claims data studies tend to overestimate the relation between antidepressant use and hip fractures. However, after correcting for such bias, a significant association persists. PMID- 15538127 TI - Enhanced heat loss and age-related hypersensitivity to diazepam. AB - Whether elderly people suffer from age-related changes in pharmacokinetics and/or pharmacodynamics with administration of benzodiazepines is still a matter of controversy. We investigated the course of brain function and thermoregulation after oral administration of a standard benzodiazepine, diazepam (DZP), in 8 healthy young men (mean age, 19.8 years; range, 18 to 23 years) and 8 healthy middle-aged and older men (mean age, 60.9 years; range, 53 to 71 years). Placebo or DZP was administered in a single-blind crossover manner to the young men (placebo, 5-mg, 10-mg DZP) and to the older men (placebo, 5-mg DZP), and plasma DZP concentration, choice reaction time, proximal body temperature, and distal body temperature were monitored with high time resolution under a modified constant routine condition to exclude masking effects. Whereas there was no evidence of age-related alterations in pharmacokinetics between the 2 groups, the older subjects, in comparison to the young subjects, showed a more delayed choice reaction time in response to the same plasma DZP level, suggesting that hypersensitivity is related to increased age. DZP at 5 mg in the older subjects induced acute and transient hypothermia to the same degree as that induced by DZP at 10 mg in the young subjects. The distal-proximal body temperature gradient (difference between distal body temperature and proximal body temperature), an indicator of blood flow in distal skin regions, showed strong positive correlation with the delay in choice reaction time in both groups. These findings suggest that hypersensitivity to benzodiazepine in older persons may be due, at least in part, to age-related changes in thermoregulation, especially in the heat loss process. PMID- 15538128 TI - Impact of the CYP2D6 ultrarapid metabolizer genotype on mirtazapine pharmacokinetics and adverse events in healthy volunteers. AB - INTRODUCTION: In vitro studies showed that biotransformation of the antidepressant drug mirtazapine is mediated by cytochrome P-450 enzymes CYP1A2, CYP2D6, and CYP3A4 with CYP2D6 contributing about 35% to total mirtazapine biotransformation. We hypothesized that ultrarapid metabolizers (defined as carriers of the CYP2D6 gene duplication plus another functional allele) have a risk for therapeutic failure due to too low tissue concentrations. METHODS: Ten healthy male volunteers carrying 1 CYP2D6 duplication allele and 1 wild-type allele, 12 carriers of 2 CYP2D6 wild-type alleles and 3 carriers of 2 functionally inactive alleles received a single dose of 45 mg racemic mirtazapine and plasma concentrations were measured from 0 to 58 hours. RESULTS: Median total clearance of racemic mirtazapine (Cl/F) was 20.1, 39.7, and 49.8 L/h in carriers of 0, 2, and 3 active genes of CYP2D6 (P = 0.002, trend test) and the median maximum plasma concentrations were 129, 159, and 76 mug/L in these 3 groups. The effects on maximal blood concentrations may indicate a contribution of CYP2D6 on mirtazapine first-pass metabolism. A trend with lower concentrations in the high activity CYP2D6 genotypes was also seen for the active metabolite desmethylmirtazapine, but without any significance. Mirtazapine concentrations showed a significant correlation with diastolic and systolic blood pressure (P = 0.05) and the correlation was even stronger when taking total mirtazapine (mirtazapine plus desmethylmirtazapine, P = 0.03), but neither blood pressure nor heart rate effects were correlated with CYP2D6 genotype. CONCLUSIONS: Consistent with the in vitro data, the genetically polymorphic enzyme CYP2D6 contributed to about 25% of total clearance in carriers of only one active allele and up to 55% in the genetically defined ultrarapid metabolizers. But the effect of the CYP2D6 gene duplication was lower than expected and high CYP2D6 activity may only explain a very small fraction of the cases with therapeutic failure in treatment with mirtazapine. PMID- 15538129 TI - Venlafaxine for the treatment of depressive episode during the course of schizophrenia. AB - The emergence of depression in the course of schizophrenia is common and arouses much interest and therapeutic concern. It has been associated with a less favorable prognosis and increased incidence of suicide. However, relatively few treatment studies have been performed in this area. The use of a combination of antidepressants and antipsychotic agents is controversial. We report an open label study carried out to evaluate the efficacy of the addition of venlafaxine in schizophrenia patients treated with antipsychotics and diagnosed with concurrent depressive episode (Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria). Patients (N = 19) who did not show spontaneous improvement after 4 weeks were assigned to a 6-week trial with add-on venlafaxine. Patients were evaluated at a 1-week interval with the Hamilton Depressive Rating Scale, the Positive and Negative Syndrome Scale, and the Clinical Global Impression Scale. All 19 patients had completed the 6-week trial. Fourteen patients (74%) showed significant improvement measured with Hamilton Depressive Rating Scale and Clinical Global Impression Scale. The mean venlafaxine dose was 146 mg/d (range: 75 to 225 mg/d). In most patients, there was a parallel decrease in psychotic symptoms. We conclude that venlafaxine may have a role in the treatment of depression in patients with schizophrenia without causing exacerbation of psychosis. PMID- 15538130 TI - Haloperidol half-life after chronic dosing. AB - In normal subjects after a single oral dose, haloperidol half-life has been reported to range 14.5-36.7 hours (or up to 1.5 days). After chronic administration, half-lives of up to 21 days have been reported. The objective of this study was to evaluate specific factors that might account for differences in haloperidol half-life in patients taking haloperidol chronically, including gender, age, weight, race, CYP2D6 and CYP3A5 genotypes, comedication, and smoking.Thirty-one patients were administered haloperidol for 4 weeks followed by a 1-week washout before administration of clozapine. Haloperidol plasma levels were measured weekly for at least 2 months after discontinuation. The geometric mean for haloperidol half-life and detectable levels duration were 3.9 and 13.8 days, respectively. Within 31 subjects, 58% (18/31) had half-lives <3 days (1.2 2.3 days) and 42% (13/31) had half-lives > or =3 days. Two of 3 patients with half-lives longer than 30 days (720 hours) and levels detectable >2 months had received haloperidol decanoate. Five patients who received haloperidol decanoate in the prior year were excluded from a comparison between patients with long haloperidol half-lives (> or =3 days, n = 10) and patients with short half-lives (<3 days, n = 16). The only significant difference between the two groups was that African-Americans (n = 4) were all found to have a long haloperidol half life (P = 0.014). CYP3A5 genotype did not appear to influence haloperidol half life but the two CYP2D6 poor metabolizer had half-lives > or =3 days. This study suggests that haloperidol half-life following repeated drug administration is substantially more prolonged than what has been observed after acute haloperidol administration. PMID- 15538131 TI - S-adenosyl-L-methionine (SAMe) as an adjunct for resistant major depressive disorder: an open trial following partial or nonresponse to selective serotonin reuptake inhibitors or venlafaxine. AB - BACKGROUND: The purpose of this open trial was to evaluate the safety, tolerability, and efficacy of oral S-adenosyl-L-methionine as an antidepressant adjunct among partial and nonresponders to serotonin reuptake inhibitors or venlafaxine. METHOD: Thirty antidepressant-treated adult outpatients with persisting major depressive disorder received 800 to 1600 mg of S-adenosyl-L methionine tosylate over a 6-week trial. RESULTS: Intent-to-treat analyses based on the Hamilton Depression Rating Scale revealed a response rate of 50% and a remission rate of 43% following augmentation with S-adenosyl-L-methionine. Gastrointestinal symptoms and headaches were the most common side effects. CONCLUSION: Augmentation of selective serotonin reuptake inhibitors or venlafaxine with S-adenosyl-L-methionine warrants a placebo-controlled trial in resistant depression. PMID- 15538132 TI - Effects of naltrexone on the subjective response to amphetamine in healthy volunteers. AB - While dopaminergic mechanisms in amphetamine-taking behavior have been extensively studied, the contribution of the endogenous opioid system is less clear. We assessed the effects of an opioid antagonist, naltrexone (50 mg), on the subjective response to an oral dose of dexamphetamine (30 mg) in 12 healthy volunteers in a double-blind, placebo-controlled design. Volunteers received a total of 4 combinations of the study preparation (placebo-naltrexone, placebo amphetamine) over 4 occasions with 1-week intervals. The primary objective of the study was to evaluate the effect of pretreatment with naltrexone on the subjective response to amphetamine. This was measured using a Visual Analog Scale, assessing the subjective effects over 7 hours. The secondary objective was to measure the effects of naltrexone on behavioral and physiologic responses to amphetamine. This was measured by blood pressure, heart rate, skin conductance, and speed of reading at the end of each session. Amphetamine produced significant effects on subjective arousal when compared to placebo after 1 hour (P < 0.001) and continued to be evident until 7 hours. Pretreatment with naltrexone significantly attenuated the subjective effects of amphetamine (P < 0.05), and this effect was time-dependent with a reduction from the 3-hour time point. Naltrexone did not influence the behavioral and physiologic effects of amphetamine in this sample. The results provide preliminary evidence that naltrexone may reduce the reinforcing effects of amphetamine via modulation of the opioid system. The potential of naltrexone as an adjunct pharmaceutical for the treatment of amphetamine dependence is promising and needs to be investigated further. PMID- 15538133 TI - Olanzapine-induced hyperglycemic hyperosmolar nonketotic coma: a case report. PMID- 15538134 TI - Reply to Dr. J-C Shim and colleagues regarding "Fluoxetine augmentation of haloperidol in chronic schizophrenia" [J Clin Psychopharmacol 2003;23:520-521]. PMID- 15538135 TI - High dose of depot risperidone in a nonresponder schizophrenic patient. PMID- 15538136 TI - Guanfacine treatment of clozapine-induced sialorrhea. PMID- 15538137 TI - Neuroleptic malignant syndrome with dermatologic complications. PMID- 15538138 TI - Absence of effect of olanzapine on primary polydipsia: results of a double-blind, randomized study. PMID- 15538139 TI - Bulimia nervosa, 5-HTTLPR polymorphism and treatment response to four SSRIs: a single-blind study. PMID- 15538140 TI - Obsessive-compulsive symptoms precipitated by methadone tapering. PMID- 15538143 TI - Brain gamma-aminobutyric acid levels are decreased in patients with phantageusia and phantosmia demonstrated by magnetic resonance spectroscopy. AB - BACKGROUND: Olfactory and gustatory hallucinations (phantosmias and phantageusias, respectively) are sensory distortions that commonly follow losses of olfactory and gustatory acuity (hyposmia and hypogeusia, respectively). The biochemical basis of these hallucinations is unclear. Functional magnetic resonance imaging has been used previously to demonstrate widespread and robust central nervous system (CNS) activation to memories of these sensory distortions in patients with these symptoms. In this study, possible CNS mechanisms responsible for these distortions were evaluated using magnetic resonance spectroscopy, because this technique has been used to measure various CNS metabolites in patients with neurologic disorders. METHODS: Forty-seven subjects were studied: 28 normal volunteers (13 men and 15 women) and 19 patients (8 men and 11 women) with persistent oral global phantageusia and/or birhinal phantosmia studied before any treatment. Four patients (1 man and 3 women) were studied before and after pharmacologic treatment that reduced the severity of their sensory distortions. All subjects were studied in a Signa 1.5-T magnetic resonance scanner with a quadrature head coil using a modified standard 2 dimensional J-point resolved excitation in the steady state (PRESS) sequence by which gamma-aminobutyric acid (GABA), glutamic acid, choline, N-acetylaspartate, and creatine (Cre) were measured in various CNS regions. Results were expressed using Cre as a denominator to determine ratios for each measurement. Differences were defined between normal subjects and patients before treatment and in patients before and after successful pharmacologic treatment. RESULTS: Before treatment, GABA levels in several CNS regions were lower in patients than in normal volunteers and were the only biochemical changes found; significantly lowered GABA levels were found in the cingulate, right and left insula, and left amygdala. No differences between patients and normal volunteers were found in any of the metabolites in the posterior occipital region. After treatment that inhibited sensory distortions, CNS GABA levels increased in the cingulate, insula, and amygdala but significantly only in the left insula and in the right and left amygdala. After this successful treatment, no change in any biochemical parameter was found in the posterior occipital region. CONCLUSIONS: These results indicate that decreased brain GABA levels can serve as biochemical markers of phantageusia and/or phantosmia in patients with these distortions and are the first biochemical changes in the CNS that reflect these sensory changes. After successful treatment of these distortions, CNS GABA levels increased to levels at or near normal, consistent with functional remission of these symptoms. These results substantiate a role for CNS GABA in the generation and inhibition of these sensory hallucinations. Although the underlying biochemical mechanism(s) for the generation of these decreased GABA levels are complex, because similar types of sensory hallucinations occur as auras or prodromata of epileptic seizure and migraine activity, these results suggest that there may be common biochemical changes among these disorders. PMID- 15538144 TI - Origin of high signal intensity in the cavernous sinus in MR angiographic source images: correlation between MR and conventional angiography. AB - OBJECTIVE: To investigate the origin of a high signal intensity in a cavernous sinus on magnetic resonance (MR) angiography. METHODS: Time-of-flight MR angiography was performed in 20 patients scheduled for conventional angiography for disease not related to carotid cavernous fistula. Venous drainage pattern was assessed, and flow rate to a cavernous sinus was semiquantitatively evaluated on conventional angiography. RESULTS: Two sinuses showed high signal intensity in the anterior portion. These were associated with direct venous drainage to the anterosuperior aspect of the cavernous sinus from the superficial middle cerebral vein, and their flow rates were among the 10% of the sides with the highest flow rates. Two other sinuses showed high signal intensity in the posterior portion. Their flow rates from the superficial petrosal sinus were also among the 10% of the sides with the highest flow rates. CONCLUSION: High signal intensity in the anterior or posterior portions of the cavernous sinus is probably a result of inflow from the superficial middle cerebral vein and the superior petrosal sinus, respectively. PMID- 15538145 TI - The added value of the apparent diffusion coefficient calculation to magnetic resonance imaging in the differentiation and grading of malignant brain tumors. AB - OBJECTIVE: ADC calculation can improve the diagnostic efficacy of MR imaging in brain tumor grading and differentiation. METHODS: Apparent diffusion coefficient (ADC) values and ratios of 33 low-grade (23 astrocytomas, 10 oligodendrogliomas) and 40 high-grade (25 metastases and 15 high-grade astrocytomas) malignant tumors were prospectively evaluated. RESULTS: Tumoral ADC values (r=-0.738, P <0.000) and ratios (r=-0.746, P < 0.000) were well correlated with higher degree of malignancy and quite effective in grading of malignant brain tumors (P < 0.000). By using cutoff values of 0.99 for tumoral ADC value and 1.22 for normalized ADC ratio, the sensitivity of MR imaging could be increased from 72.22% to 93.75% and 90.63%, the specificity from 81.08% to 92.68% and 90.24%, PPV from 78.79% to 90.91% and 87.88%, and NPV from 75.00% to 95.00% and 92.50%, respectively. CONCLUSION: ADC calculation was quite effective in grading of malignant brain tumors but not in differentiation of them and added more information to conventional contrast-enhanced MR imaging. PMID- 15538146 TI - Comparison of perfusion computed tomography with diffusion-weighted magnetic resonance imaging in hyperacute ischemic stroke. AB - OBJECTIVE: In this study, perfusion CT and diffusion-weighted magnetic resonance imaging (DWI) were compared as means of assessing the ischemic brain in hyperacute stroke. METHODS: Twenty patients with ischemic stroke underwent perfusion computed tomography (CT) and magnetic resonance imaging (MRI) studies <3 hours after stroke onset. Cerebral blood flow thresholds were used to delineate the ischemic lesion, penumbra, and infarct. Correlations between the volume of the hypoperfused areas, the abnormality volume in admission DWI and follow-up CT/MRI studies, and the clinical National Institutes of Health Stroke Scale (NIHSS) scores were performed. RESULTS: The volume of the ischemic (core and penumbra) lesion on admission perfusion CT was correlated with the volume of admission DWI abnormalities (r=0.89, P=0.001). The infarcted core tissue volume (on admission CT) correlated more strongly (r=0.77, P=0.0001) than the admission DWI abnormality volume (r=0.69, P=0.002) with the follow-up infarct volume on fluid-attenuated inversion recovery images. A correlation was demonstrated between infarct volume in perfusion CT and follow-up DWI abnormality volume (r=0.89, r=0.77, P=0.002). Significant correlations were found between ischemic and infarct region volumes in perfusion CT and NIHSS admission and follow-up scores (P < or = 0.01). CONCLUSIONS: Both imaging modalities provide a sufficient assessment of the hyperacute brain infarct, with significant correlation between them and the clinical condition at admission. Perfusion CT allows differentiation of the penumbra and infarct core region with significant predictive value of follow-up infarct volume and clinical outcome. PMID- 15538147 TI - Diffusion-weighted MR imaging in Japanese encephalitis. AB - OBJECTIVE: This study was performed to evaluate the role of diffusion-weighted imaging (DWI) in the diagnosis of Japanese encephalitis (JE) and to look for any relationship of the apparent diffusion coefficient (ADC) values with duration of the illness. METHODS: We performed DWI in fourteen patients of JE. T2 weighted (T2W) and DWI were compared for number and location of lesions in all patients. Based on imaging patients were divided in three groups: group 1 (n=9) showing more lesions on DWI compared with T2W images, group 2 (n=3) with equal number of lesions on T2W and DWI and group 3 (n=2) with lesions more pronounced on T2W than DWI. ADC values were computed for all the lesions. The time interval between onset of neurologic signs/symptoms and MRI were charted and correlated with ADC values. RESULTS: DWI was helpful in making early diagnosis of JE by showing characteristic involvement of bilateral thalami in four patients. Nine out of fourteen patients showed additional lesions on DWI. ADC from lesions in groups I, II and III measured 0.648 +/- 0.099 x 10 mm/s, 0.739 +/- 0.166 x 10 mm/s and 1.123 +/- 0.185 x 10 mm/s respectively. The ADC from the lesions in group 1 was significantly lower compared with group 2 (P value <0.05) while it was higher in group 3 lesions compared with the other two groups. There was a significant direct correlation of ADC values with the disease duration in these cases (r=0.847, P <0.01). CONCLUSIONS: DWI is helpful in early diagnosis and characterization of the duration of the lesions in JE. PMID- 15538148 TI - Prenatal magnetic resonance imaging of rhombencephalosynapsis and associated brain anomalies: report of 3 cases. AB - Rhombencephalosynapsis (RES) is a rare congenital abnormality characterized by vermian agenesis and fusion of the cerebellar hemispheres. In the 3 cases reported here, prenatal magnetic resonance imaging clearly depicted cerebellar features characteristic of RES, allowing a definite diagnosis. PMID- 15538149 TI - Solitary pulmonary nodules: a comparative study evaluated with contrast-enhanced dynamic MR imaging and CT. AB - OBJECTIVE: The aim of this prospective study was to compare the diagnostic performances of dynamic MR imaging and CT for the differentiation of benign and malignant solitary pulmonary nodules (SPNs). METHODS: Eighty-one patients with SPNs (32 malignant, 49 benign) underwent dynamic MR imaging (n=31), dynamic CT (n=27), or both (n=23). The degree of peak enhancement of benign and malignant SPNs was compared on both dynamic MR imaging and CT. Receiver operating characteristic (ROC) analysis was performed to compare the diagnostic performances of dynamic MR imaging and CT. RESULTS: The malignant SPNs revealed significantly greater degrees of peak enhancement on dynamic MR imaging (mean +/- SD [p%SI] 131.2 +/- 46.1 versus 54.2 +/- 45.3; range [p%SI] 82.6-260.0 versus 0.7-171.7; P <0.0001) and CT (mean +/- SD [DMI] 37.8 +/- 15.1 versus 17.9 +/- 21.8; range [DMI] 14.1-68.2 versus -5.4-107.6; P=0.0004). Although dynamic MR imaging was somewhat superior to dynamic CT, the diagnostic performances of the 2 modalities based on ROC analysis were not statistically significant. CONCLUSIONS: Dynamic MR imaging and CT seem to be equally well suited for the differentiation between benign and malignant SPNs. PMID- 15538150 TI - Amyloidosis and lymphoproliferative disease in Sjogren syndrome: thin-section computed tomography findings and histopathologic comparisons. AB - OBJECTIVE: To describe the thin-section computed tomography (CT) findings of Sjogren syndrome accompanying pulmonary amyloidosis and lymphoproliferative disease and to compare these with histopathologic findings. SUBJECTS AND METHODS: The thin-section CT findings of 5 women (age range: 42-59 years, mean age=50 years) with primary Sjogren syndrome accompanying pulmonary amyloidosis and lymphoproliferative disease were reviewed retrospectively by 2 chest radiologists, and decisions on findings were reached by consensus. The pathologic specimens of parenchymal lesions (nodules, dense consolidation, and cystic lesion at CT) obtained using video-assisted thoracoscopic surgery were studied to compare with the thin-section CT findings. RESULTS: Nodules, observed in all 5 patients, were variable in size and ranged from 3 to 24 mm (mean=9.9 mm) in diameter, with lobulated or irregular margins. Nodular calcifications were present in 3 patients. Cysts, which also were observed in all patients, ranged from 4 to 45 mm (mean=18.6 mm) in diameter, with a thin (1-2 mm) or no visible wall. Multiple cysts were observed, especially in the distal portion of narrowed bronchioles. Nodules and cysts showed a random distribution. Mild bronchial wall thickening with bronchial dilatation was seen in all patients, ground-glass opacities were seen in 3, and consolidation was seen in 1. Nodules, consolidation, and bronchial wall thickening at CT were caused histopathologically by the interstitial and peribronchiolar deposition of mixed amyloid and lymphoproliferative cells. Cysts lined with respiratory epithelium contained amyloid deposition and lymphoproliferative cells in their walls. CONCLUSION: Sjogren syndrome accompanying pulmonary amyloidosis and lymphoproliferative disease manifests as multiple, large, thin-walled cysts; multiple nodules; parenchymal opacity; and bronchiectasis. These findings are caused by the interstitial or peribronchial infiltration of mixed amyloid and lymphoproliferative cells. PMID- 15538151 TI - Primary mediastinal lymphoma: characteristic features of the various histological subtypes on CT. AB - OBJECTIVE: To assess the characteristic features of the primary mediastinal lymphoma (PML) on CT and to test the relationship between CT findings and the likelihood of the 3 most common subtypes (Hodgkin lymphoma [HL], mediastinal diffuse large B-cell lymphoma [Med-DLBCL], and precursor T-cell lymphoblastic lymphoma [T-LBL]). METHODS: Sixty-six consecutive patients with pathologically proven PML including 29 patients with HL, 21 with Med-DLBCL, and 16 with T-LBL underwent CT prior to therapy. CT scans were independently reviewed by 2 radiologists who were blinded to the pathologic diagnosis for the following considerations: pattern of involvement (i.e., morphologic features, mass size, and contrast enhancement pattern), and ancillary findings at other sites including neck, abdomen, and pelvis. Interobserver agreement was measured by Kappa statistics, and independent predictors were calculated using multiple logistic regression analysis for determining the likelihood of the subtypes based on CT. RESULTS: Characteristic features of HL included irregular contour of the anterior mediastinal mass (20 of 29, 69%) and high prevalence of associated mediastinal lymphadenopathy (28 of 29, 97%). Characteristic features of Med-DLBCL included regular contour (14 of 21, 67%) and absence of cervical and abdominal lymphadenopathy (0 of 21). Characteristic features of T-LBL included regular contour (12 of 16, 75%) and high prevalence of cervical (9 of 16, 56%) and abdominal (6 of 16, 38%) lymphadenopathy and splenomegaly (11 of 16, 69%). CT findings independently associated with increased likelihood of HL were surface lobulation (P <0.01), the absence of vascular involvement (P <0.01), or pleural effusion (P <0.05). The presence of vascular involvement was associated with increased likelihood of Med-DLBCL (P <0.001). Furthermore, CT findings including the presence of cervical lymph nodes or inguinal lymph nodes (P <0.001), the presence of pericardial effusion (P <0.05), and the absence of surface lobulation (P <0.05) were significantly associated with the likelihood of T-LBL. CONCLUSION: The various histologic subtypes of PML have characteristic manifestations in the neck, chest, and abdomen, which allow their distinction on CT. PMID- 15538152 TI - Severe acute respiratory syndrome: thin-section computed tomography features, temporal changes, and clinicoradiologic correlation during the convalescent period. AB - OBJECTIVE: To evaluate thin-section computed tomography findings of patients with severe acute respiratory syndrome (SARS) in the convalescent period and to correlate the results with clinical parameters and lung function tests. METHODS: Ninety-nine severe acute respiratory syndrome patients with persistent changes on follow-up chest radiography were included. One hundred seventy computed tomography examinations at baseline (n=70), 3 months (n=56), and 6 months (n=44) were retrospectively evaluated to determine the extent of ground-glass opacification, reticulation, and total parenchymal involvement. Patients' demographic information, clinical information during treatment, and results of lung function tests at 3 and 6 months were correlated with computed tomography findings. RESULTS: A significant serial improvement in the extent of overall ground-glass opacification, overall reticulation, and total parenchymal involvement was observed (P <0.01). Advanced age, previous intensive care unit admission, mechanical ventilation, alternative treatment, higher peak lactate dehydrogenase, and peak radiographic involvement during treatment showed a positive correlation with overall reticulation and total parenchymal involvement at 6 months. There was a significant negative correlation between overall reticulation and total parenchymal involvement with diffusion capacity adjusted for hemoglobin at 3 and 6 months (P <0.01). CONCLUSION: Lung changes on thin section computed tomography of severe acute respiratory syndrome patients improved with time during the convalescent period and showed a significant correlation with advanced age, parameters indicating severe illness, and diffusion capacity adjusted for hemoglobin on follow-up. PMID- 15538153 TI - Pulmonary dirofilariasis: computed tomography findings and correlation with pathologic features. AB - OBJECTIVE: The purpose of this study was to describe the computed tomography (CT) and pathologic features of 5 nodules of pulmonary dirofilariasis in 4 patients. METHODS: Four patients with 5 nodules of pathologically confirmed pulmonary dirofilariasis who under went CT were enrolled, and the imaging interpretations were retrospectively compared with the histopathologic characteristics. RESULTS: Three of the 4 patients had a solitary nodule, and the remaining patient had 2 nodules. All the nodules were distributed in the right lower lobe and were attached to the pleura. They were all round or oval in shape and ranged in size from 11 to 22 mm in largest diameter (mean=17 mm). On thinner section CT, the nodules had a well-defined smooth margin with or without a shallow notch; they were connected to the arterial branch and, occasionally, to the venous branch. On contrast-enhanced CT, all the nodules contained a homogeneous low-attenuation area, which corresponded to areas of coagulative necrosis on histopathologic examination. CONCLUSION: Although the CT findings of a pulmonary dirofilariasis nodule are nonspecific, awareness of the findings on contrast-enhanced CT and the pathologic appearance of this rare benign condition may facilitate its differentiation from a malignant nodule. PMID- 15538154 TI - Tracheobronchopathia osteochondroplastica in a patient with silicosis: CT, bronchoscopy, and pathology findings. AB - A case of tracheobronchopathia osteochondroplastica in a patient with silicosis is reported, showing a rare association of disease. Etiological hypotheses and clinical aspects are discussed. Radiologic, bronchoscopic, and pathologic findings are demonstrated with emphasis on the role of computed tomography (CT) in the diagnosis of this disease. PMID- 15538155 TI - Hemodynamic and morphologic changes after portal vein embolization: differential effects in central and peripheral zones in the liver on multiphasic computed tomography. AB - OBJECTIVE: To evaluate hemodynamic and morphologic effects in the liver after portal vein embolization (PVE). METHODS: Hepatic computed tomography scans of 7 patients who had undergone preoperative PVE were retrospectively reviewed. Pre- and post-PVE computed tomography densities were evaluated for the unenhanced, late arterial, and portal venous phases in peripheral and central hepatic regions and in the 3 main hepatic veins. Relative changes in areas in these regions were assessed in 5 evaluable patients with serial post-PVE scans. RESULTS: During the late arterial phase, enhancement was significantly higher after PVE than it was before PVE in the peripheral hepatic regions, and it was higher in the peripheral regions than in the central regions. Enhancement was also significantly higher in the right main hepatic vein than in the middle and left hepatic veins during the late arterial phase. The ratio of areas of the peripheral/central regions decreased significantly after PVE. CONCLUSIONS: Zonal enhancement in the late arterial phase changed after PVE and seemed to be associated with differential parenchymal atrophy. We speculate that the hepatic arterial supply increases peripherally and that peribiliary/periportal plexuses maintain the portal supply centrally. PMID- 15538156 TI - Computed tomography imaging of gastrointestinal stromal tumors with pathology correlation. AB - Gastrointestinal stromal tumors (GISTs) are mesenchymal tumors that typically arise in association with the muscularis propria of gastrointestinal (GI) tract wall. They occur most frequently in the stomach (60%) but also can occur in the small bowel (30%) or elsewhere, including the colon and rectum (5%) and esophagus (<5%). In addition, a GIST may occur as a primary tumor of the omentum, mesentery, or retroperitoneum. They account for 1%-3% of gastric neoplasms, 20% of small bowel tumors, and 0.2%-1% of colorectal tumors. These tumors arise pathologically from the wall of the GI tract and can be characterized as benign, borderline, or with low or high malignant potential based on the pathologic appearance. Computed tomography (CT) plays an important role for the diagnosis and staging of these neoplasms because it can identify the tumor and assess for local spread or distant metastases. This article reviews the role of CT in the detection and characterization of GISTs. The optimal CT technique for visualization of small bowel tumors is reviewed, and a variety of GISTs are illustrated. Pathologic correlation is also included. PMID- 15538157 TI - Differentiation of adrenal adenomas from metastases with unenhanced computed tomography. AB - OBJECTIVE: To investigate whether the diagnostic accuracy of unenhanced computed tomography (CT) regarding the differentiation of adrenal adenomas from adrenal metastases is increased by applying a combination of morphologic criteria instead of only measuring the density values of the tumor. PATIENTS AND METHODS: Unenhanced CT scans of 56 patients with an adrenal mass and a history of an extra adrenal malignancy were analyzed for size, attenuation, contour, and structure characteristics of the adrenal tumor. Coefficients yielded by multiple logistic regression analysis were used for the construction of an additive total score (score S) that included several diagnostic criteria. The reliability of the total score and all parameter combinations was tested by receiver operating characteristic (ROC) analysis. The nature of the adrenal lesion was determined by follow-up CT (40 patients), percutaneous biopsy (15 patients), or surgery (1 patient). Twenty-four of the neoplasms were adenomas, and 32 were found to be metastases. RESULTS: The score of the combined CT parameters showed the largest area under the ROC curve. The highest predictive power indicated by the model was calculated at a cutoff value of 7.05, with a sensitivity of 100% and a specificity of 96.8% for the detection of metastases. At 6.85 points as the cutoff value, the scoring system still maintained a sensitivity of 95.8% and a specificity of 96.9%. CONCLUSION: The differentiation between adrenal adenomas and metastases is improved by applying our scoring system compared with any single parameter alone. The total score is obtained by adding 10% of the density values to the size in centimeters, plus 2 if the contour of the lesion is blurred and plus 1 if the structure is inhomogeneous. By setting the threshold at 7 points, all but 1 lesion were classified correctly. PMID- 15538158 TI - Evaluation of renovascular hypertension: comparison of functional MRI and contrast-enhanced MRA with a routinely performed renal scintigraphy and DSA. AB - OBJECTIVE: The main objective of this study was to evaluate a clinically suspected renovascular hypertension (RVH) by means of MRI. It was to find out if functional MRI (fMRI) is able to provide adequate results with regard to MAG3 captopril scintigraphy and if contrast-enhanced MR angiography (MRA) is able to provide adequate results in the stenosis grading compared with the nonselective digital subtraction angiography (DSA). METHODS: This open, monocentric, prospective, phase 3 study included patients with a clinically suspected RVH. For fMRI a dynamic TurboFLASH sequence and for MRA a single-shot breath-hold flash 3D sequence was performed. Gadodiamide was injected as contrast medium. RESULTS: Sixty patients were included in the study. The correlation between fMRI and scintigraphy had an accuracy, a sensitivity, and a specificity of 69%, 5%, and 92%, respectively, and correlations between MRA and DSA of 95%, 92%, and 96%. CONCLUSION: The noninvasive MRA can replace DSA in the diagnosis of renal artery stenosis. However, fMRI can not replace renal scintigraphy. PMID- 15538159 TI - Computed tomography features of portal hypertensive gastropathy. AB - OBJECTIVE: The aim of this study was to detect the computed tomography features in portal hypertensive gastropathy (PHG). METHODS: This study included 32 patients with portal hypertension. Ten of 32 patients were diagnosed with (PHG), and the other 22 patients were not diagnosed with PHG on the basis of upper endoscopic examination. Dynamic computed tomography studies were performed in all 32 patients. Each computed tomography scan was evaluated with regard to whether the gastric inner layer was enhanced or not. RESULTS: Nine patients with PHG had delayed enhancement on the inner layers of gastric walls, but in 17 of 22 patients without PHG, delayed enhancement was not observed. CONCLUSION: Enhancement on the inner layer of gastric walls may reflect gastric congestion. Portal hypertensive gastropathy should be suspected when this finding is detected on computed tomography scans in patients with portal hypertension. PMID- 15538160 TI - Magnetic resonance imaging of ovarian cancer arising in endometriomas. AB - The magnetic resonance imaging (MRI) findings in 2 cases of ovarian cancer arising in an endometrioma are presented. In both cases, the cancer was evident as an enhancing mass within a blood-filled adnexal cyst. The MRI findings of an enhancing mass within a blood-filled adnexal cyst should prompt consideration of ovarian cancer arising in an endometrioma, facilitating recognition of a superimposed malignancy and potentially allowing for earlier and more efficacious treatment. PMID- 15538161 TI - A new pitfall on abdominal PET/CT: a retained surgical sponge. AB - Positron emission tomography (PET)/computed tomography (CT) and multidetector CT findings of an abdominal retained surgical sponge (RSS) are presented. A PET/CT study performed for evaluation of a suspected abdominal tumor demonstrated the inconclusive finding of a hypometabolic area surrounded by increased 2-[fluorine 18] fluoro-2-deoxy-D-glucose uptake. Follow-up contrast-enhanced multidetector CT suggested the correct diagnosis of an RSS. This is the first known report of the PET/CT appearance of an RSS. PMID- 15538163 TI - Primary cardiac lymphoma demonstrated by delayed contrast-enhanced magnetic resonance imaging. AB - Delayed contrast-enhanced magnetic resonance (MR) imaging that nullifies the signal of normal myocardium produces great differences in myocardial signal intensity between normal and infarcted myocardium. A case of primary cardiac lymphoma is presented in which delayed contrast-enhanced MR imaging clearly identified the localization and extension of a lymphoma infiltrating the myocardium. PMID- 15538162 TI - Evaluation of the quantitative capability of a high-resolution positron emission tomography scanner for small animal imaging. AB - OBJECTIVE: The quantitative capability of a positron emission tomography scanner for small animal imaging was evaluated in this study. METHODS: The microPET P4 (Concorde Microsystems, Knoxville, TN) scanner's capability for dynamic imaging and corrections for radioactive decay, dead time, and attenuation were evaluated. Rat brain and heart studies with and without attenuation correction were compared. A calibration approach to convert the data to nanocuries per milliliter was implemented. Calibration factors were determined using calibration phantoms of 2 sizes with and without attenuation correction. Quantitation was validated using the MiniPhantom (Data Spectrum, Chapel Hill, NC) with hot features (5:1 ratio) of different sizes (4, 6.4, 8, 13, and 16 mm). RESULTS: The microPET P4 scanner's ability to acquire dynamic studies and to correct for decay, dead time, and attenuation was demonstrated. The microPET P4 scanner provided accurate quantitation to within 6% for features larger than 10 mm. Sixty percent of object contrast was retained for features as small as 4 mm. CONCLUSIONS: The microPET P4 scanner can provide accurate quantitation. PMID- 15538165 TI - Anomalous origin of a posterior descending artery from the right pulmonary artery: report of a rare case diagnosed by multidetector computed tomography angiography. AB - The case of a mildly symptomatic 43-year-old man with evidence of reversible inferior wall ischemia by radioisotope stress testing is described. Invasive catheterization showed abnormal connections between the coronary arteries and the pulmonary circulation. Multidetector computed tomography elucidated the previously undescribed finding of a posterior descending artery originating from the right pulmonary artery. PMID- 15538164 TI - Cardiac lipoma and lipomatous hypertrophy of the interatrial septum: cardiac magnetic resonance imaging findings. AB - Cardiac lipomas are uncommon, usually asymptomatic benign primary tumors of the heart that may incidentally be discovered during computed tomography (CT) or magnetic resonance imaging (MRI). Although the finding of a low-attenuation mass with density similar to fat on CT is pathognomonic, the MRI appearances of fatty cardiac masses are variable depending on the sequences employed. The MRI findings of 4 patients with cardiac lipomas or lipomatous hypertrophy of the interatrial septum are presented. All patients had lesions with characteristic low-signal intensity margins and high central signal intensity on "bright-blood" balanced gradient-echo cardiac MRI sequences with very low repetition and echo times. It is proposed that this appearance results from intravoxel phase cancellation effects occurring at the fat/tissue interface and is sufficiently characteristic to obviate the need for confirmatory CT. PMID- 15538166 TI - Potential MR signs of recurrent carpal tunnel syndrome: initial experience. AB - OBJECTIVE: In nonoperated patients, the MR diagnosis of carpal tunnel syndrome (CTS) is difficult. In the postoperative patient this difficulty is compounded. Consequently, we sought to evaluate for potential MR signs of postoperative CTS. METHODS: At 1.5 T, 41 wrists in 37 patients with previous CTS release were evaluated by two observers for 1) flexor retinacular regrowth; 2) median nerve: a) high T2 signal, b) proximal enlargement, c) fibrous fixation, d) neuroma, and e) entrapment; 3) flexor tenosynovitis; 4) mass, bursitis, accessory muscle, distal belly progression, or excessive deep fat; 5) hamate fracture; and 6) volar nerve migration. Electromyography (EMG), operative findings, and clinical follow up were used to determine the presence of recurrent CTS. RESULTS: Fifteen of 41 wrists had recurrent CTS. Retinacular regrowth was seen in 4/15 (27%) with and 7/26 (27%) without recurrent CTS (P=0.7). Excessive fat was seen in 1/15 (7%) with and 2/26 (8%) without CTS (P=0.19). No patient had incomplete resection of flexor retinaculum, scarring, neuroma of nerve, or tendon laceration; bursitis, accessory or distal muscle progression of muscle belly, or hamate fracture. Nerve edema with high T2 signal was seen in 4/15 (27%) with and 3/26 (12%) without CTS (P=0.16); proximal enlargement was seen in 6/15 (40%) with CTS and 2/26 (8%) without CTS (P=0.007). Also, 1 patient with recurrent disease demonstrated a mass and 1 other patient without CTS had nerve entrapment. Tenosynovitis was seen in 9/15 (60%) with and 9/26 (35%) without recurrent CTS (P=0.02). Counterintuitively, the nerve was more palmar with recurrent CTS than without (mean 6.9/8.9 mm). CONCLUSION: Only proximal enlargement, tenosynovitis, and the rare mass may help to diagnose recurrent CTS by MR. However, there appears to be a subgroup of patients with recurrent neuropathy related to an excessively superficial median nerve. PMID- 15538167 TI - Sectional neuroanatomy of the face. AB - Needle electromyography and the injection procedures of the face are difficult because of the small size and close anatomic relations of the facial structures. Local injections of the face have increased in frequency since the introduction of botulinum toxin. This article intends to provide anatomically accurate schematics of innervation of the muscles of facial expression that can be used to interpret magnetic resonance images of the muscles and nerves. Cross-sectional schematics of the face were drawn as they appear in imaging projections. The relevant cranial and spinal nerves were color coded. The muscles and skin surfaces were labeled and assigned the color of the appropriate nerves. An organized comprehensive map of the motor innervation of the face allows the physician to increase the accuracy and efficacy of interventional procedures. This could also assist the electromyographer in correlating the clinical and electrophysiologic findings with magnetic resonance images. PMID- 15538170 TI - Can autophagy protect against neurodegeneration caused by aggregate-prone proteins? AB - Protein conformation disorders or proteinopathies are a growing family of human diseases that are characterized by the accumulation of proteins in intracellular aggregates (also known as inclusions) in specific tissues/organs. The role of aggregates in these diseases has been a subject of vigorous debate. However, irrespective of the nature(s) of the toxic species, it is desirable for cells to be able to control the levels of these toxic proteins and restrict their accumulation. Here we discuss how the autophagy-lysosome pathway may regulate protein clearance in some of the protein conformation disorders and why this pathway may represent a possible therapeutic target in such conditions. PMID- 15538171 TI - FMRP and its target RNAs: fishing for the specificity. AB - Learning and memory difficulties observed in patients with fragile X syndrome, as well as in a mouse model for the syndrome, are partially due to impaired translational regulation of neuronal mRNAs encoding key molecules for the synaptic structure and function. There has been intense interest in characterizing the mRNAs that are regulated by the fragile X mental retardation protein (FMRP) in the neuronal cell. A large number of candidate FMRP-interacting mRNAs has been identified over the last few years and three models have been described so far to explain the specificity of these interactions. Here, we report our vision on how they could work in the same and/or in different pathways and suggest that the three mechanisms may not be mutually exclusive. PMID- 15538172 TI - GLAST expression on bullfrog Muller cells is regulated by dark/light. AB - Using immunofluorescence double-labeling, Western blotting and confocal laser scanning microscopy, we investigated if and how the expression of glial high affinity glutamate/aspartate transporter (GLAST) in bullfrog Muller cells may be regulated by dark/light. Compared with light-adapted retinas, the expression of GLAST in Muller cells was overall up-regulated in retinas dark-adapted for 30 min but declined in retinas dark-adapted for longer (>30 min) periods. The declined expression level of GLAST during prolonged dark adaptation was raised by immersion with 1 mM glutamate. These results suggest that glutamate uptake mediated by GLAST could be regulated dynamically and efficiently in accord with dark/light-induced changes in glutamate release of retinal neurons. PMID- 15538173 TI - Auditory M50 and M100 responses to broadband noise: functional implications. AB - The functional significance of the M50 and M100 auditory evoked fields remains unclear. Here we report auditory evoked field data from three different studies employing wide-band noise stimuli. We find that, for the same stimuli, the strength of the M100, as well as its lateralization, are task-modulated. The M50, in contrast, shows three properties: It is dramatically more pronounced for noise stimuli than for pure tones, does not seem to be task dependent, and, is significantly stronger in the left hemisphere in all task conditions. These contrasting patterns of activation shed light on the properties of the response generating mechanisms and suggest roles in the process of auditory figure-ground segregation. PMID- 15538174 TI - Neural network for encoding immediate memory in phonological processing. AB - The aim of this fMRI study was to identify neuroanatomical substrates of immediate memory underlying phonological processing. To distinguish encoding of immediate memory from rehearsal, participants were required to match tones from the first and last positions of a three-syllable list to their following probes in an immediate-recognition paradigm. The first position task included intervening distractors between the target and probe. Increased activations were found in the left inferior frontal gyrus, right lateral cerebellum, and medial frontal gyrus for the target tone in first position. This network mediates articulatory encoding in immediate-response, and articulatory rehearsal in delayed-response paradigms. These findings support a working memory model in which rehearsal is optional, while encoding is an obligatory component of the phonological loop. PMID- 15538175 TI - Expression of kinin B1 receptors in the spinal cord of streptozotocin-diabetic rat. AB - Previous studies have reported cardiovascular and nociceptive responses after intrathecal injection of kinin B1 receptor (B1R) agonists in the model of streptozotocin (STZ)-diabetic rat (diabetic). The aim of this study was to measure the early up-regulation of B1R binding sites and mRNA in the thoracic spinal cord of diabetic and control rats. Data show significant increases of specific B1R binding sites in the dorsal horn of diabetic rats 2 days (+315%), 7 days (+303%) and 21 days (+181%) after STZ treatment. Levels of mRNA were significantly increased (+68%) at 2 and 7 days but not at 21 days. These data bring the first molecular evidence for an early up-regulation of B1R in the spinal cord of diabetic rat. PMID- 15538177 TI - Does familiarity facilitate the cortical processing of music sounds? AB - Automatic cortical sound discrimination, as indexed by the mismatch negativity (MMN) component of the auditory evoked potential, is facilitated for familiar speech sounds (phonemes). In musicians as compared to non-musicians, an enhanced MMN has been observed for complex non-speech sounds. Here, musically trained subjects were presented with sequences of either familiar (tonal) or structurally matched unfamiliar (atonal) triad chords, both with either fixed or randomly transposed chord root pitch. The MMN elicited by deviant chords did not differ for familiar and unfamiliar triad sequences, and was undiminished even to unfamiliar deviant sounds which were consciously undetectable. Only subsequent attention-related components indicated facilitated cognitive processing of familiar sounds, corresponding to higher behavioral detection scores. PMID- 15538176 TI - DTI and impulsivity in schizophrenia: a first voxelwise correlational analysis. AB - Compromised white matter (WM) integrity in inferior frontal WM has been related to impulsivity in men with schizophrenia. However, these relationships may be more widespread. Fractional anisotropy (FA) derived from diffusion tensor imaging of 25 men with schizophrenia was transformed into Talairach space. Correlations between FA and impulsiveness were examined on a voxelwise basis. We found negative correlations between FA and impulsivity in inferior frontal WM, anterior cingulate, caudate, insula, and inferior parietal lobule. Positive correlations were obtained in the left postcentral gyrus, right superior/middle temporal gyrus, and bilateral fusiform gyrus. These areas may comprise a fronto-temporo limbic circuit that modulates impulsivity. The voxelwise correlation method can serve as a hypothesis-generation method for relating target behaviors to their underlying neural networks. PMID- 15538178 TI - Altered vulnerability to kainate excitotoxicity of transgenic-Cu/Zn SOD1 neurones. AB - The neurotoxicity of the AMPA/kainate receptor agonist kainate was investigated in motor and cortical neurones from mice over-expressing the wild-type and G93A mutant form of Cu/Zn superoxide dismutase (SOD1) human gene, a mouse model of familial amyotrophic lateral sclerosis. G93A mutant motor neurones were more vulnerable and wild-type SOD1 motor neurones were more resistant to kainate toxicity than were controls. Voltage-gated Na channels blockage prevented G93A mutant SOD1 motor neurone death. Cortical cultures exhibited fewer differences in their vulnerability to kainate toxicity. These results demonstrate that SOD1 over expression selectively affects the sensitivity to kainate excitotoxicity of motor neurones but not neocortical neurones, and that wild-type SOD1 expression increases the resistance to excitotoxicity of motor neurones. PMID- 15538179 TI - Striatal preprotachykinin gene expression reflects parkinsonian signs. AB - Striatal preprotachykinin (PPT) gene expression was measured in MPTP-treated cats when symptomatic and during various stages of recovery from parkinsonism using in situ hybridization histochemistry. Animals expressing severe (1 week post-MPTP) or moderate (3 weeks post-MPTP) parkinsonian sensorimotor deficits had significantly reduced striatal PPT mRNA expression. In contrast, fully recovered animals (6 weeks post-MPTP) had striatal PPT mRNA levels that were not significantly different from normal. Thus, PPT gene expression in the striatum appears to reflect presence or absence of sensorimotor deficits in MPTP-treated cats. PMID- 15538180 TI - Quantitative analysis of tooth pulp afferent terminals in the rat brain stem. AB - This study analyzed quantitatively the ultrastructural features of tooth pulp afferent terminals and their presynaptic axonal endings (p-endings) in the trigeminal principal (Vp), dorsomedial oral (Vdm), and caudal nuclei (Vc). Mitochondrial volume, active zone area, apposed surface area, and vesicle number were highly correlated with afferent bouton volume. The afferent bouton volume varied widely in Vp, compared to that in Vdm and Vc. The values of all parameters of p-endings were within a narrow range, and were smaller than those of afferent boutons. The afferent bouton volume correlated with the number of postsynaptic dendrites and p-endings. These results suggest that pulpal afferent information is regulated in a unique manner in the each trigeminal sensory nucleus. PMID- 15538181 TI - Direct current stimulation over MT+/V5 modulates motion aftereffect in humans. AB - While there is strong evidence for the central role of the human MT+/V5 in motion processing, its involvement in motion adaptation is still the subject of debate. We used transcranial direct current stimulation (tDCS) to test whether MT+/V5 is part of the neural network involved in the long-term adaptation-induced motion after-effect in humans. It was found that both cathodal and anodal stimulation over MT+/V5 resulted in a significant reduction of the perceived motion after effect duration, but had no effect on performance in a luminance-change-detection task used to determine attentional load during adaptation. Our control experiment excluded the possibility that the observed MT+/V5 stimulation effects were due to a diffused modulation of the early cortical areas, i.e. by the stimulation applied over MT+/V5. These results provide evidence that external modulation of neural excitability in human MT+/V5 affects the strength of perceived motion after-effect and support the involvement of MT+/V5 in motion adaptation processes. PMID- 15538182 TI - BDNF regulates NMDA receptor activity in developing retinal ganglion cells. AB - Brain-derived neurotrophic factor (BDNF) modulates glutamate receptors of the NMDA type in many areas of the brain. We assessed whether BDNF exerts an effect on NMDA receptor properties in retinal ganglion cells during early postnatal development. Electrophysiological responses to the glutamate agonist NMDA (500 microM-2 mM) in retinal slices of wildtype and BDNF deficient mice (bdnf-/-) were recorded using the whole-cell patch-clamp technique. Retinal ganglion cells of bdnf-/- mice displayed significantly smaller NMDA currents than those of age matched wildtype mice. Remarkably, NMDA receptor activity was restored by incubating retinal slices of bdnf-/- mice in BDNF (50 ng/ml) for 1-3 h. We suggest that BDNF plays a role in the activation of functional NMDA receptors in early ganglion cell development. PMID- 15538183 TI - Neuregulin potentiates agrin-induced acetylcholine receptor clustering in myotubes. AB - Agrin and neuregulin (HRG-beta1) play complementary roles in synapse formation. While HRG-beta1 induces transcriptional up-regulation of postsynaptic proteins, here we present evidence that it can potentiate agrin-induced acetylcholine receptor (AChR) clustering in C2 myotubes. Agrin induced maximal AChR clustering in 4 h. HRG-beta1 treatment for 4 h produced no increase over basal AChR cluster numbers. When myotubes were treated for 4 h with 100 pM agrin, HRG-beta1 augmented AChR cluster numbers by 2-fold compared to myotubes treated with 100 pM agrin alone. Thus, HRG-beta1 can potentiate agrin-induced AChR clustering. PMID- 15538184 TI - Differential expression patterns of TrkB ligands in the macaque monkey brain. AB - We measured the concentrations of brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4) in the brains of mature and developing macaque monkeys. In the adult macaque brain, the highest level of BDNF was found in the hippocampus. Comparisons between cortical regions showed higher levels of BDNF in the prefrontal, temporal, and parietal association cortices than elsewhere. In contrast, NT-4 levels were 3-30 times lower than those of BDNF. During development, the contents of BDNF and NT-4 protein in cerebral cortices were highest at the postnatal second month and at embryonic day 140, respectively. These results suggest that the two TrkB ligands, BDNF and NT-4, may be differently regulated and might play separate roles in monkey cortical development. PMID- 15538185 TI - Inflammatory mediators and growth factors in the spinal cord of G93A SOD1 rats. AB - Amyotrophic lateral sclerosis (ALS) is a devastating motor neuron disease. One mechanism involved in ALS pathology is neuroinflammation. Neuroinflammation is mediated by soluble pro-inflammatory molecules such as cytokines, prostaglandins and nitric oxide. Studies on transgenic mice demonstrated the expression of pro inflammatory mediators in early stages of murine ALS. Recently a transgenic rat model became available. Since species differences in regard to cytokine expression have been reported in other disease models we set out to validate the neuroinflammatory hypothesis in the ALS-transgenic rat. We investigated the expression of inflammatory mediators and growth factors in the spinal cord by semi-quantitative RT-PCR. We found that several pro-inflammatory mediators are up regulated at asymptomatic and end-stages, whereas VEGF, a neuroprotective factor was down-regulated. PMID- 15538186 TI - Neuregulin genotype and medication response in Finnish patients with schizophrenia. AB - Neuregulin 1 is involved both in neurodevelopment and neurotransmitter mechanisms in the brain. There is evidence of an association between neuregulin 1 genotype and schizophrenia. We compared neuregulin 1 genotypes in patients with schizophrenia (n=94) and control subjects (n=395) of Finnish origin by using one SNP (SNP8NRG221533) as a genetic marker. We also analyzed NRG1 genotype with regard to age at onset and between responders and non-responders to conventional antipsychotics. The NRG1 genotype or allele frequencies showed similar distributions between patient and control groups. Age at onset was not associated with NRG1 genotype. The TT genotype was overrepresented in the non-responders group compared with the responders (p=0.013). Further studies are needed to ascertain the significance of neuregulin genotype in medication response to schizophrenia. PMID- 15538187 TI - Neuroprotection by encapsulated choroid plexus in a rodent model of Huntington's disease. AB - Choroid plexus from neonatal pigs was encapsulated in alginate microcapsules and transplanted into the rat striatum. Three days later, the same animals received unilateral injections of quinolinic acid (225 nmol) into the ipsilateral striatum. Choroid plexus transplants ameliorated the weight loss and motor impairments resulting from QA. Histological analysis demonstrated that choroid plexus transplants reduced the volume of striatal damage and protected ChAT-, but not NADPH-diaphorase-positive neurons. These data are the first to demonstrate that transplanted choroid plexus cells can protect striatal neurons from excitotoxic damage and that this strategy may ultimately prove relevant for the treatment of Huntington's disease. PMID- 15538188 TI - Amnesia produced by pre-training infusion of serotonin into the substantia nigra. AB - Interference with dopaminergic, GABAergic or cholinergic activity of the substantia nigra is sufficient to disrupt cognitive functions. In spite of the known interactions between these neurotransmitters and serotonin, there is a notable scarcity of data germane to the participation of nigral serotonin in learning and memory. It was important, therefore, to investigate the possible involvement of serotonin in cognition. Pre-training injections of serotonin into the nigra of rats produced strong amnesia of an aversively-motivated task (inhibitory avoidance) compared to similar injections into the cerebral cortex and zona incerta. The present findings support the hypothesis that nigral serotonergic activity is involved in memory functions. PMID- 15538189 TI - Hormone replacement therapy and age-related brain shrinkage: regional effects. AB - Neuroprotective properties of estrogen have been established in animal models, but clinical trials of hormone replacement therapy (HRT) produced contradictory results. We examined the impact of HRT on age-related regional changes in human brain volume. Six brain regions were measured twice, five years apart, in 12 healthy women who took HRT and in matched controls who did not. The controls showed a typical pattern of differential brain shrinkage in the association cortices and the hippocampus with no change in the primary visual cortex. In contrast, women who took HRT showed comparable shrinkage of the hippocampus but no significant shrinkage of the neocortex. Future large scale studies may benefit from applying regional rather than global measures in assessment of brain integrity. PMID- 15538190 TI - Reduced electrodermal response to errors predicts poor sustained attention performance in attention deficit hyperactivity disorder. AB - Research into attention deficit hyperactivity disorder (ADHD) has indicated abnormalities in electrodermal system activity (EDA) and separately, deficits in sustained attention. Here we asked whether reduced EDA in ADHD was consequential for the attention problems associated with this disorder. On a sustained attention task ADHD participants had higher overall error rates and exhibited a marked decrease in skin conductance responses (SCRs) to errors, relative to control children. Further, sustained attention errors were predicted by SCR amplitudes. Evidence of post-error slowing in both groups suggested comparable error awareness. It is proposed that attenuation of the normal autonomic response to errors reflects impairment in ADHD participants' psychological response to error significance and results in decreased behavioural correction and hence poorer sustained attention. PMID- 15538191 TI - Opposing BOLD responses to reciprocated and unreciprocated altruism in putative reward pathways. AB - Mesencephalic dopamine neurons are believed to facilitate reward-dependent learning by computing errors in reward predictions. We used fMRI to test whether this system was activated as expected in response to errors in predictions about whether a social partner would reciprocate an act of altruism. Nineteen subjects received fMRI scans as they played a series of single-shot Prisoner's Dilemma games with partners who were outside the scanner. In both ventromedial prefrontal cortex and ventral striatum, reciprocated and unreciprocated cooperation were associated with positive and negative BOLD responses, respectively. Our results are consistent with the hypothesis that mesencephalic dopamine projection sites carry information about errors in reward prediction that allow us to learn who can and cannot be trusted to reciprocate favors. PMID- 15538192 TI - Letters lost in space: hemispace dependent handwriting errors. AB - Although handwriting is a daily life activity commonly attributed to the left hemisphere in the majority of right-handers, it is also known to require attentional and spatial mechanisms that rely on right hemispheric processing. The underlying spatial organization of handwriting in patients with right brain damage remains unresolved. Here we show in a patient with circumscribed right superior parietal damage that handwriting systematically depends on the hand's position in space with respect to her body-midline. Most importantly, handwriting in contralesional space not only leads to spatial but also to language errors. This suggests that the right hemisphere's role in handwriting may surpass its generally assumed purely spatial contribution. We discuss our results in term of co-registration between both cerebral hemispheres in language processing. PMID- 15538193 TI - The role of the right dorsolateral prefrontal cortex in visual change awareness. AB - Recently, the neural correlates of change detection vs change blindness have been investigated using fMRI. Results revealed that conscious perception of change is associated with enhanced activity in a neural network comprising the parietal (bilateral) and right dorsolateral prefrontal (DLPF) cortex. Here, by means of repetitive transcranial magnetic stimulation (rTMS), we unveil the causal role of the right DLPF cortex in perceiving changes. When rTMS was applied to this area, change perception was impaired as compared to left DLPF rTMS and sham stimulation. This result is important as it shows, for the first time, that conscious change perception is associated with normal activity in the right DLPF cortex. Our findings are in agreement with a recent view emphasizing the role of frontal areas, in addition to classical ventral and dorsal pathways, in visual awareness. PMID- 15538194 TI - Eye gaze cueing facilitates neural processing of objects in 4-month-old infants. AB - A major issue in developmental science is how infants use the direction of other's eye gaze to facilitate the processing of information. Four-month-old infants passively viewed images of an adult face gazing toward or away from objects. When presented with the objects a second time, infants showed differences in a slow wave event-related potential, indicating that uncued objects were perceived as less familiar than objects previously cued by the direction of gaze of another person. This result shows that the direction of eye gaze of another cannot only bias infant attention, but also lead to enhanced information processing of the objects concerned. PMID- 15538195 TI - Gender related differences on the EEG during a simulated mobile phone signal. AB - The present study investigated the gender-related influence of electromagnetic fields (EMF), similar to that emitted by mobile phones, on brain activity. Ten women and nine men performed a short memory task (Wechsler test), both without (baseline) and with exposure to a 900 MHz signal. The EEG energy of the total waveform and the alpha, beta, delta and theta; rhythms were calculated from the recordings of 15 scalp electrodes. Baseline EEG energy of males was greater than that of females, while exposure to EMF decreased EEG energy of males and increased that of females. Memory performance was invariant to EMF exposure and gender influences. These findings indicate that EMF may exert a gender-related nfluence on brain activity. PMID- 15538196 TI - Error-related negativity reflects detection of negative reward prediction error. AB - Error-related negativity (ERN) is a negative deflection in the event-related potential elicited in error trials. To examine the function of ERN, we performed an experiment in which two within-participants factors were manipulated: outcome uncertainty and content of feedback. The ERN was largest when participants expected correct feedback but received error feedback. There were significant positive correlations between the ERN amplitude and the rate of response switching in the subsequent trial, and between the ERN amplitude and the trait version score on negative affect scale. These results suggest that ERN reflects detection of a negative reward prediction error and promotes subsequent response switching, and that individuals with high negative affect are hypersensitive to a negative reward prediction error. PMID- 15538199 TI - Jay Y. Gillenwater. PMID- 15538200 TI - Urology at the University of Virginia. PMID- 15538201 TI - Prospective outcomes associated with migration from preoperative to intraoperative planned brachytherapy: a single center report. AB - PURPOSE: Brachytherapy (BT) for prostate cancer can be performed with either preoperative (PO) or intraoperative (IO) planned dosimetry. Potential benefits of intraoperative include fewer procedures for the patient, cost savings and improved accuracy of seed implantation leading to improved tumor control and urinary side effect profile. We report our experience with transition to IO planned BT after performing more than 600 PO planned implants since 1997. MATERIALS AND METHODS: From September 2001 to February 2003, 46 consecutive patients with T1-3N0M0 adenocarcinoma of the prostate underwent BT. IO dosimetry was performed in 23 patients, while PO dosimetry was used in 23 immediately before changing to IO. American Urological Association (AUA) symptom index questionnaires were administered preoperatively and postoperatively. All patients underwent postoperative dosimetry by computerized tomography. Total, irritative and obstructive AUA scores were compared in the 2 groups using analysis of covariance. Models were adjusted for pretreatment variables of symptom scores, type of procedure and time since procedure. RESULTS: Median followup was 47 and 45 days for PO and IO dosimetry, respectively. No differences were observed in seed, needle numbers or prostate size in the 2 groups. Average operative times were higher (47 vs 79 minutes, p <0.01) in the IO group but they decreased to nearly the same levels as PO implants in the first 23 cases so treated. Slopes of operative time over date of procedure differed significantly between methods (p <0.01). Comparing PO to IO dosimetry adjusted estimate of difference was -1.96 for total (95% CI -6.4, 2.5), -0.48 for obstructive (95% CI -3.3, 2.3) and -1.78 for irritative (95% CI -3.9, 0.31) AUA score. These differences were neither statistically nor clinically significant. CONCLUSIONS: Our experience indicates that intraoperative planned BT is easily implemented in clinical practice as a result of a short learning curve. In addition, the approach is not associated with any changes in early postoperative voiding symptoms and, due to only marginally longer operative times, may have a cost advantage by eliminating the preplanning visit and ultrasound. PMID- 15538202 TI - Dendritic cell based vaccines: progress in immunotherapy studies for prostate cancer. AB - PURPOSE: No effective treatment is currently available for metastatic prostate cancer. Dendritic cell (DC) based cancer vaccine research has emerged from the laboratories to human clinical trials. We describe progress in the development of DC based prostate cancer vaccine. MATERIALS AND METHODS: The literature was reviewed for major contributions to a growing number of studies that demonstrate the potential of DC based immunotherapeutics for prostate cancer. Background topics relating to DC based immunotherapy theory and practice are also addressed. RESULTS: DCs have been recognized as the most efficient antigen presenting cells that have the capacity to initiate naive T cell response in vitro and in vivo. During their differentiation and maturation pathways, dendritic cells can efficiently capture, process and present antigens for T cell activation. These characteristics make DC an attractive choice as the cellular adjuvant for cancer vaccines. Advances in DC generation, loading, and maturation methodologies have made it possible to generate clinical grade vaccines for various human trials. More than 100 DC vaccine trials, including 7 studies of patients with advanced prostate cancer have been reported to date. These vaccines were generally well tolerated with no significant adverse toxicity reported. Clinical responders have been identified in these studies. CONCLUSIONS: The new prospects opened by DC based vaccines for prostate cancer are fascinating. When compared to conventional treatments, DC vaccinations have few side effects. Improvements in patient selection, vaccine delivery strategies, immune monitoring and vaccine manufacturing will be crucial in moving DC based prostate cancer vaccines closer to the clinics. PMID- 15538203 TI - Adjunctive therapy for men with high risk localized and locally advanced prostate cancer: targeting disseminated tumor cells. AB - PURPOSE: Our research into the pathophysiology of micrometastatic dissemination and cancer recurrence has resulted in the initiation of a clinical trial for men with clinically localized and locally advanced disease. MATERIALS AND METHODS: We describe the development of this trial, which exploits anti-angiogenesis therapy, and delineate how our understanding of prostate cancer metastasis influenced its design. RESULTS: Prostate cancer is a heterogeneous disease. Although many men can be cured with local therapy, a large majority with clinically localized disease will experience a relapse usually at a distant site. This result is most likely due to micrometastatic dissemination early in the disease process. Therefore, successful contemporary treatment of many men with prostate cancer should include a combination of local and systemic therapies. Fortunately, cellular, molecular and genetic features that may predict which men are most in need of this therapeutic approach are being identified and characterized. This insight not only supports the rationale for a combination therapeutic approach to prostate cancer management, but will help identify the pathways and agents that provide the most promising targets for intervention. CONCLUSIONS: Despite advances in prevention and early detection, refinements in surgical technique, and improvements in radiation and systemic therapies, the ability to cure all men with prostate cancer remains unattainable. The continuing challenge is the successful eradication of recurrent and metastatic disease. PMID- 15538204 TI - A comparison of minimally invasive open and laparoscopic radical retropubic prostatectomy. AB - PURPOSE: A cohort of more than 1,000 patients undergoing open minimally invasive radical retropubic prostatectomy (mini-lap RRP, 8.0 cm incision) is compared to published series of laparoscopic radical retropubic prostatectomy (RRP). MATERIALS AND METHODS: A cohort of 209 consecutive patients undergoing mini-lap RRP at Emory University Hospital was evaluated. Characteristics noted were patient age, comorbidity status, preoperative prostate specific antigen, operative time, perioperative complications, blood transfusions, pathological staging, potency and continence. Postoperative analgesia was analyzed in a cohort of 30 consecutive recent patients. These data were compared to published reports of similar perioperative characteristics in laparoscopic RRP series. RESULTS: Differences between the cohorts of patients reviewed were found in age, preoperative prostate specific antigen, operative time and rate of blood transfusion. The series were comparable in comorbidity status, postoperative complications, rates of incontinence, rates of impotence and margin positivity. Analgesic use was quantified in a small group of patients according to the amount of narcotic analgesia required during the postoperative hospital stay. CONCLUSIONS: Mini-lap RRP can generally be performed in less time and at less cost than most laparoscopic radical prostatectomies. It can be performed with regional anesthesia and has excellent postoperative results compared to contemporary laparoscopic procedures. Mini-lap RRP requires less training than laparoscopic RRP. With its minimal morbidity, mini-lap RRP remains an attractive technique that can easily be adopted by the experienced retropubic surgeon. PMID- 15538205 TI - Radical prostatectomy with preservation of urinary continence: pathology and long term results. AB - PURPOSE: A continence sparing radical retropubic prostatectomy is described and evaluated. Results are compared with those of patients who underwent a standard anatomical radical retropubic prostatectomy previously by the same surgeon. MATERIALS AND METHODS: A total of 275 consecutive patients underwent a continence sparing radical prostatectomy by a single surgeon (DLG) between 1996 and 2003. The technique is described in detail. RESULTS: Total continence (no pad or device) was found immediately upon catheter removal in 36% of patients, within 14 days in 69% and within 7 weeks in 78%, compared to 1%, 6% and 41% of 80 patients who underwent the standard operation. Median time to total continence was 1 day for the new group versus 63 days for the standard surgery group. Positive margins were found in 6.9% of the new group versus 11% of the first 80 patients. Of the former patients 80% have a prostate specific antigen of less than 0.2 ng/ml at an average followup of 12.5 years. With the new procedure 90% of patients have a prostate specific antigen of less than 0.2 ng/ml with an average followup of 5.2 years. There was no operative mortality or unusual complication. The typical patient was discharged home 1 to 3 days postoperatively. CONCLUSIONS: Preservation of the continence mechanism at the level of the bladder neck and proximal prostatic urethra results in earlier return of continence without adversely affecting cancer control. It is a relatively simple way to improve surgical results. PMID- 15538206 TI - Laparoscopic partial nephrectomy techniques: developments and translation. AB - PURPOSE: Laparoscopic partial nephrectomy has recently emerged as a potential surgical option for select renal masses. Several new techniques and devices that may aid in laparoscopic partial nephrectomy are reviewed. MATERIALS AND METHODS: I review several techniques studied and/or developed in our laboratory. Each technique was evaluated for effectiveness in the porcine model and is in translation to clinical practice. RESULTS: Three techniques are reviewed. A hand assisted approach incorporating renal hilar clamping with hypothermia has proven successful for complex and multifocal lesions. Recent clinical studies, and our laboratory and clinical experience have shown a saline cooled monopolar dissector to be a valuable adjunct. A new and simple technique of achieving rapid hypothermia using a pure laparoscopic approach is described. CONCLUSIONS: Laparoscopic partial nephrectomy continues to develop as a standard of care for select renal masses. New devices and techniques will continue to make the procedure safer and reproducible. PMID- 15538207 TI - Chronic prostatitis: Charlottesville to Seattle. AB - PURPOSE: Since few men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) have culturable bacteria by traditional approaches, we used sensitive molecular methods to determine presence of fastidious microorganisms. MATERIALS AND METHODS: We evaluated 135 men with CP/CPPS by standardized clinical evaluation, and by lower tract localization cultures and chamber counts of expressed prostatic secretions of leukocytes. We excluded from study patients with bacteriuria, bacterial prostatitis, urethritis or positive urethral cultures. Prostate biopsy was obtained using a double-needle technique to limit contamination. We chose molecular approaches because previous studies had used culture antigen detection in urine, urethral swabs and expressed prostatic secretions. However, interpretation of such studies is complicated because urogenital samples often acquire bacteria while passing through the urethra. We used specific and broad-spectrum polymerase chain reaction (PCR) assays. RESULTS: Only 10 (8%) of the 135 subjects had positive specific PCR assays, including Mycoplasmia genitalium, Chlamydia trachomatis and Trichomonas vaginalis. Our findings suggested that C. trachomatis, T. vaginalis and M. genitalium may be identified in some patients with CP/CPPS, even among men with no evidence of urethritis and with negative urethral cultures and other assays. The broad spectrum PCR assays provided the most provocative findings. DNA encoding tetracycline resistance was identified in 25% of subjects, and 77% of subjects had evidence of 16S rDNAs. The white blood cell concentration in the prostatic secretions correlated with identification of 16S rDNAs in prostate tissue (p <0.01). CONCLUSIONS: Delineating the precise role of these organisms in the etiology of CP/CPPS may help define better diagnostic and treatment algorithms. PMID- 15538208 TI - Interstitial cystitis, chronic nonbacterial prostatitis and chronic pelvic pain syndrome in men: a common and frequently identical clinical entity. AB - PURPOSE: In this ongoing review we characterize interstitial cystitis in men, identifying multiple clinical factors in the diagnosis and treatment of men with interstitial cystitis. MATERIALS AND METHODS: An ongoing and retrospective chart review of 92 men with interstitial cystitis within our large single specialty practice was performed. Presenting complaints, duration of symptoms, epidemiology, and clinical and physical findings were reviewed. RESULTS: All patients met National Institute for Diabetes and Digestive and Kidney Diseases criteria for the diagnosis of interstitial cystitis. Of the patients 83% were white and 17% were American Indians of Cherokee descent. Average patient age at diagnosis was 51 years and average duration of symptoms was 8.8 years. The most commonly reported initial complaints were mild suprapubic discomfort, which rapidly progressed in an average of 21/2 years to severe suprapubic discomfort, severe dysuria and severe urinary frequency. Sexual dysfunction occurred in 60% of patients with painful ejaculation. The most common physical findings were suprapubic tenderness to palpation and anterior rectal wall tenderness on digital rectal examination. Of the patients 40% had microscopic hematuria. All patients demonstrated severe glomerulations under anesthesia with cystoscopy and hydraulic distention of the bladder. Average bladder volume was 510 cc. CONCLUSIONS: Interstitial cystitis in males appears to be more common than historically reported. Interstitial cystitis in males and patients with chronic pelvic pain syndrome and chronic nonbacterial prostatitis share many clinical findings. A higher incidence of interstitial cystitis had been found in American Indian males of Cherokee descent and deserves further investigation. PMID- 15538209 TI - Pathophysiology of unilateral ureteral obstruction: studies from Charlottesville to New York. AB - PURPOSE: More than 30 years ago the decreased renal blood flow and glomerular filtration rate characteristic of unilateral ureteral obstruction (UUO) was described. In the ensuing time, much has been learned about the involvement of nitric oxide (NO) and transforming growth factor-beta (TGF-beta) in the pathophysiology of UUO. MATERIALS AND METHODS: We measured renal blood flow and glomerular filtration rate in dogs and rats, and assessed the effect of altering the availability of NO on these parameters. In rats and mice we used an antibody to TGF-beta, NO synthase gene deletion and inducible nitric oxide synthase gene therapy to assess the role of TGF-beta and NO in renal fibrosis. RESULTS: Results of our studies suggest 2 strategies that have the potential to be incorporated into clinical trials. The first would be replenishment of the NO system with arginine (or a NO donor) to decrease renovascular resistance and increase renal nitric oxide. Either in addition to the first strategy or separately, interstitial fibrosis could be targeted. Strategies for inhibiting fibrosis include antibody to TGF-beta, use of antisense oligonucleotides to TGF-beta, use of drugs that inhibit other pro-fibrotic mediators or gene therapy to inhibit fibrosis. CONCLUSIONS: While these studies have primarily centered on acute UUO, the findings in this model of renal injury may potentially be transferable to other entities that are characterized by decreased renal function and increased renal fibrosis such as different forms of interstitial nephropathy or diabetic glomerulopathy. PMID- 15538210 TI - Corticotropin releasing factor: a mediator of emotional influences on bladder function. AB - PURPOSE: We review evidence linking overactive bladder (OAB) and interstitial cystitis with anxiety and depression, and highlight the importance of corticotropin releasing factor (CRF) as a potential mediator of these conditions. MATERIALS AND METHODS: We review the literature and describe ongoing experiments. RESULTS: CRF is expressed in areas of the central nervous system that control voiding and response to stress. CRF is increased during anxiety, depression and pain as well as functional disorders of the pelvic viscera. Epidemiological studies reveal an association between anxiety and voiding disorders. Experiments in our laboratory show that CRF stimulates bladder activity while CRF antagonists inhibit OAB. CONCLUSIONS: Epidemiological, genetic and neuropharmacological evidence links disorders of voiding and anxiety. CRF may participate in these events and warrants further investigation as a therapeutic target in the treatment of OAB and interstitial cystitis. PMID- 15538211 TI - The molecular pathology of experimental testicular torsion suggests adjunct therapy to surgical repair. AB - PURPOSE: We review the work of our laboratory in discovering the pathophysiological mechanisms that underpin testicular response to testicular torsion. Evidence from animal models is used to discover pathways that might be amenable to manipulation by therapeutic regimens. MATERIALS AND METHODS: Rats and mice were subjected to 1 and 2 hours of testicular torsion, respectively. Preliminary experiments determined that those are the times of torsion in those species that produce severe testicular atrophy and germ cell apoptosis. A variety of biochemical and molecular biological techniques were used to determine the mechanism(s) leading to spermatogenic disruption and germ cell apoptosis. RESULTS: Testicular torsion can eliminate spermatogenesis despite return blood flow, continued Sertoli cell function and perhaps the continued production of testosterone by Leydig cells, although the latter point is not completely resolved. Torsion repair is followed by a period of germ cell apoptosis, accumulation of testicular neutrophils and increased testicular oxidative stress. Testicular vascular E-selectin expression is increased after torsion repair as are a number of cytokines important to the recruitment of neutrophils. Elements of the c-Jun-N-terminal kinase pathway are important in this process. The presence of neutrophils leads to intratesticular oxidative stress, and oxidative stress has been significantly reduced by intravenous infusion of oxygen radical scavengers at the time of torsion repair. CONCLUSIONS: Testicular torsion causes loss of spermatogenesis and a significant increase in germ cell apoptosis due to an increase in testicular oxidative stress concomitant with reperfusion. Oxidative stress arises with recruitment of neutrophils, and the recruitment of neutrophils occurs due to E-selectin expression on the surface of the testicular venules after torsion repair. The cytokines, tumor necrosis factor-alpha and interleukin-1beta, activate the stress related kinase pathway to E-selectin expression after torsion repair. Oxidative stress is relieved by infusion of oxygen radical scavengers, which results in a significant salvage of testicular function. PMID- 15538214 TI - The roles of myofibroblasts in prostate carcinogenesis. PMID- 15538215 TI - Future of laparoscopy. PMID- 15538216 TI - Man versus machine: brave new surgery. PMID- 15538217 TI - Transplantation. PMID- 15538218 TI - Health related quality of life assessments for patients with bladder cancer. AB - PURPOSE: We review the current data regarding the impact of various therapeutic alternatives for bladder cancer on health related (HR) quality of life (QOL). MATERIALS AND METHODS: A MEDLINE search of the English literature from 1966 to November 2003 using the search terms "cystectomy," "intravesical therapy" or "bladder cancer" and quality of life yielded 33 articles. Studies of metastatic disease were excluded. The method of evaluation of quality of life in each article was assessed, particularly the instruments used and the population studied. RESULTS: The majority of these reports (20 of 33) compared the impact of different types of urinary diversion on quality of life. The reports provided conflicting data, with several studies showing no significant difference in overall QOL. Some studies revealed a greater improvement in HRQOL with time following continent diversion. Only 2 reports discussed evaluation of HRQOL during intravesical therapy. Either an established standardized QOL instrument or an internally validated instrument was used in all but 8 of the studies. No reports compared the HRQOL effects of bladder sparing treatments to radical cystectomy. CONCLUSIONS: There is no single measure of HRQOL predominantly used in patients with bladder cancer. There is a lack of data comparing HRQOL outcomes in patients receiving bladder sparing therapies versus cystectomy. Development of a universally applicable validated instrument would allow a more effective comparison of HRQOL outcomes in these patients as well as in those with superficial and muscle invasive bladder cancer. PMID- 15538219 TI - The diagnosis and treatment of osteoporosis in men on androgen deprivation therapy for advanced carcinoma of the prostate. AB - PURPOSE: Androgen deprivation therapy (ADT) for advanced prostate cancer increases the risk of osteoporosis. Thus, the practicing urologist should be aware of the appropriate assessment and management. In this article we review the tests designed to diagnose osteoporosis as well as treatment options. We also review methods to monitor the response to therapy and make recommendations for management. MATERIALS AND METHODS: We performed a MEDLINE (1966 to 2004) search for the terms male, osteoporosis, bone mineral density, prostate cancer, androgen deprivation therapy, bisphosphonates, estrogen and the combinations thereof. We then constructed a management algorithm based on the best evidence available. RESULTS: Dual energy x-ray absorptiometry of the hip is the gold standard test for osteoporosis. Biochemical markers of bone turnover are not suitable for diagnosis but they have been shown to be useful for monitoring the response to treatment. Smoking cessation, weight bearing exercise, and vitamin D and calcium have been shown to help improve bone mineral density (BMD). Bisphosphonates have been demonstrated to increase BMD and decrease fracture risk in men with osteoporosis. Estrogens have also recently been shown to decrease bone turnover and increase BMD in men on ADT. CONCLUSIONS: Hip dual energy x-ray absorptiometry should be performed in all men who are anticipated to be on long-term ADT. In addition, all men on ADT should receive vitamin D and calcium supplementation, and perform regular weight bearing exercise. The value of smoking cessation cannot be overstated. In men who have osteopenia or osteoporosis bisphosphonate therapy should be initiated. Estrogen therapy has shown promise but specific recommendations cannot be made at this time. PMID- 15538220 TI - A systematic review and critique of the literature relating hospital or surgeon volume to health outcomes for 3 urological cancer procedures. AB - PURPOSE: We performed a systematic review and critique of the literature of the relationship between hospital or surgeon volume and health outcomes in patients undergoing radical surgery for cancer of the bladder, kidney or prostate. MATERIALS AND METHODS: Four electronic databases were searched to identify studies that describe the relationship between hospital or surgeon volume and health outcomes. RESULTS: All included studies were performed in North America. A total of 12 studies were found that related hospital volume to outcomes. For radical prostatectomy and cystectomy all 8 included studies showed improvement in at least 1 outcome measure with increasing volume and never deterioration. For nephrectomy the 4 included studies produced conflicting results. Four studies were found that related surgeon volume to outcomes. All radical prostatectomy and cystectomy studies showed that some outcomes were better with higher surgeon volume and never deterioration. We did not find any studies of the effect of surgeon volume on outcomes after nephrectomy. The 3 studies of the combined effect of hospital and surgeon volume on outcomes after radical prostatectomy or cystectomy suggest that high volume hospitals have better outcomes, in part because of the effect of surgeon volume and vice versa. CONCLUSIONS: Outcomes after radical prostatectomy and cystectomy are on average likely to be better if these procedures are performed by and at high volume providers. For radical nephrectomy the evidence is unclear. The impact of volume based policies (increasing volume to improve outcomes) depends on the extent to which "practice makes perfect" explains the observed results. Further studies should explicitly address selective referral and confounding as alternative explanations. Longitudinal studies should be performed to evaluate the impact of volume based policies. PMID- 15538221 TI - Impact of cyclooxygenase-2 gene expression on tumor invasiveness in a human renal cell carcinoma cell line. AB - PURPOSE: We have previously confirmed that cyclooxygenase-2 (COX-2) is expressed in a human renal cell carcinoma (RCC) cell line and it has an important role in cell tumorigenesis and angiogenesis. In the current study we evaluated the impact on cell adhesion and tumor invasiveness in human RCC cell lines by transfection of COX-2 sense and antisense cDNAs. MATERIALS AND METHODS: A human RCC cell line that expresses COX-2 was transfected with COX-2 sense or antisense cDNA. E cadherin expression in parental cells of OS-RC-2 and transfectants was detected by real-time polymerase chain reaction and Western blotting. The expression of beta-catenin was detected by Western blotting. Zymography was used to detect gelatinase activity. CD44 expression in parental cells and transfectants was detected by fluorescence activated cell sorting. Cell adhesion was detected by adhesion assay and cell invasive ability was detected by invasion assay. RESULTS: E-cadherin expression was increased in antisense transfectants and decreased in sense transfectants compared with parental cells at the mRNA and protein levels. However, obvious consistent changes in beta-catenin expression could not be confirmed in parental cells and transfectants, nor were there any significant differences in gelatinase activity in parental cells and transfectants. CD44 expression was increased in sense transfectants and decreased in antisense transfectants compared with parental cells. Adhesion to hyaluronan coated wells was significantly enhanced in sense transfectants and inhibited in antisense transfectants compared with parental cells. Compared with parental cells invasive ability was significantly increased in sense transfectants and decreased in antisense transfectants. CONCLUSIONS: The results demonstrate that COX-2 expression has a crucial role in cell invasion ability and the suppression of COX 2 expression might regulate adhesion molecule expression and inhibit invasive ability in the RCC cell line OS-RC-2. PMID- 15538222 TI - Renal cell carcinoma MIB-1, Bax and Bcl-2 expression and prognosis. AB - PURPOSE: Proliferation and programmed cell death (apoptosis) are key factors in oncogenesis and tumor progression. In carcinogenesis important regulators of apoptosis are members of the Bcl-2 family. In this family the Bcl-2 gene has an inhibitory effect on apoptosis, while Bax promotes cell death. In renal cell carcinoma (RCC) the associations between Bcl-2 proteins and RCC prognosis have been controversial. We evaluated Bax and Bcl-2 levels in RCC, and their associations with prognosis, proliferation and traditional prognostic factors. MATERIALS AND METHODS: Our prospective study population comprised 138 consecutive patients who underwent radical nephrectomy for RCC. Immunostaining and semiquantitative indices for Ki-67 (MIB-1), Bax and Bcl-2 were estimated. Their associations with prognosis were explored. RESULTS: On univariate analysis according to survival statistically significant differences were achieved by Bax (positive vs negative HR 3.04, 95% CI 1.27 to 7.23), Bcl-2 (positive vs negative HR 0.43, 95% CI 0.23 to 0.81), MIB-1 (continuous HR 1.03, 95% CI 1.001 to 1.064), Fuhrman nuclear class (4 vs 1 plus 2 HR 8.15, 95% CI 3.13 to 21.20) and stage (4 vs 1 HR 60.04, 95% CI 13.99 to 257.68). Only stage (HR 47.96, 95% CI 10.85 to 212.03) and Fuhrman classification (HR 4.32, 95% CI 1.60 to 11.65) attained statistical significance on Cox regression multivariate analysis. CONCLUSIONS: In our prospective study Bax and Bcl-2 showed a statistically significant association with prognosis in RCC but did not achieve the status of independent prognostic factors. Further studies are needed to clarify the role of the apoptotic process in tumor progression and prognosis. PMID- 15538223 TI - The membrane proteases adams and hepsin are differentially expressed in renal cell carcinoma. Are they potential tumor markers? AB - PURPOSE: ADAMs (a disintegrin and metalloproteinases) as cell surface proteins with adhesion and protease activity, and hepsin as a transmembrane protease have important roles in many biological processes. We assessed the expression of various ADAMs and of hepsin in human renal cell carcinoma (RCC), and correlated expression with clinicopathological data. MATERIALS AND METHODS: mRNA expression of ADAM-8, 17, 19, 28, TS1 and TS2, and of hepsin was investigated in paired tissue samples from cancerous and noncancerous parts of the kidneys of 27 patients with RCC who underwent tumor nephrectomy. Measurements were performed by quantitative real-time reverse transcriptase-polymerase chain reaction on a LightCycler instrument (Roche Applied Science, Mannheim, Germany). The data were related to housekeeping gene porphobilinogen deaminase mRNA. RESULTS: All ADAMs except ADAM-TS1 were significantly higher but hepsin was less expressed (at least p <0.05) in cancerous vs matched noncancerous tissue. Expression was differentially related to tumor stage. ADAM-8 and ADAM-TS2 over expression as well as decreased hepsin expression were associated with shorter patient survival. The Cox proportional hazards regression model revealed that ADAM-TS2 was an independent prognostic factor for cancer related death. ADAM-8 was the best predictor of distant metastases. CONCLUSIONS: The differential expression of hepsin and ADAMs suggests early and late involvement of membrane proteases in the development of RCC. Their association with the clinical outcome illustrates their potential usefulness as biomarkers for RCC. PMID- 15538224 TI - Symptoms as well as tumor size provide prognostic information on patients with localized renal tumors. AB - PURPOSE: T stage stratification of organ confined renal tumors is based only on tumor size. Currently T1a and T1b are defined as tumors less or greater than 4 cm. However, to our knowledge the validity of this stratification has not been determined. We determined whether symptoms could add additional prognostic information when integrated with tumor size into the TNM classification. MATERIALS AND METHODS: Patients with T1-T2N0M0 renal tumors at 6 academic centers in Europe and the United States were included in this study. T stage was defined according to the 2002 TNM classification. Age, gender, T stage, tumor size, symptoms at presentation, Fuhrman grade and cancer specific survival were determined in all cases. Survival estimates were compared using the Kaplan-Meier method and multivariate analysis of the data were performed with the Cox model. RESULTS: A total of 1,771 patients with pT1-T2N0M0 renal tumors were included in this study. There were 1,148 males and 623 females. Mean age was 59.6 years. Median tumor size was 5 cm. Of the tumors 781 (44.1%), 616 (34.8%) and 374 (21.1%) were stages T1a, T1b and T2, respectively. In 825 patients (46.6%) symptoms were related to renal cancer. T stage and symptoms strongly correlated, in that 67%, 51% and 29% of patients with T1a, T1b and T2 tumors, respectively, were asymptomatic. Symptoms increased the risk of cause specific death for each T stage level. On multivariate analysis Fuhrman grade (HR 1.46), T stage (HR 1.81) and symptoms (HR 2.98) were independent predictors of survival. Based on these results 4 groups resulting from combinations of 2002 TNM stage and symptoms with significantly different risks of death were defined, namely 1) T1a-4 cm or less without symptoms, 2) T1b-4 cm or less with symptoms and greater than 4 cm without symptoms, 3) T2a-greater than 4 cm and 7 cm or less with symptoms, and 4) T2b greater than 7 cm with symptoms CONCLUSIONS: In this study we noted that a system combining tumor size and symptoms can accurately stratify patients for predicting survival in those with organ confined renal tumors. Our data support the idea that symptoms should be integrated in further modifications of the TNM system. PMID- 15538225 TI - Laparoscopic radical nephrectomy for large (greater than 7 cm, T2) renal tumors. AB - PURPOSE: Laparoscopic radical nephrectomy has emerged as a standard of care in appropriate candidates with clinical stage T1 renal tumors (7 cm or less). Herein we present our experience with laparoscopic radical nephrectomy for clinical stage T2 tumors (greater than 7 cm). MATERIALS AND METHODS: Patients undergoing laparoscopic radical nephrectomy between September 1997 and July 2003 were retrospectively subdivided into group LAPT1-166 with tumor size 7 cm and group LAPT2-65 with tumor size greater than 7 cm. Also, group LAPT2 was compared with a group of 34 contemporary, comparable patients undergoing open radical nephrectomy for tumor greater than 7 cm (group OPENT2). RESULTS: Compared with group LAPT1, group LAPT2 had younger patients, larger tumors and greater blood loss (100 vs 200 ml) (each p <0.001). Importantly operative time, analgesic requirements, hospital stay, and convalescence and complication rates were comparable. Group LAPT2 and group OPENT2 patients had similar sized tumors (9.2 and 9.9 cm, respectively) but shorter operative time (p = 0.03), lesser blood loss (p <0.001), shorter hospital stay (p <0.001) and more rapid convalescence (p = 0.02) occurred in LAPT2. CONCLUSIONS: Laparoscopic radical nephrectomy for stage T2 renal masses (greater than 7 cm) is feasible and efficacious. Laparoscopic nephrectomy offers the advantages of decreased blood loss, shorter hospital stay and more rapid recovery over open radical nephrectomy for comparable tumors greater than 7 cm. Although surgical outcomes are comparable with laparoscopic radical nephrectomy for smaller tumors (7 cm or less), adequate laparoscopic experience is necessary before performing radical nephrectomy for large T2 tumors. PMID- 15538226 TI - Bladder cancer as a prognostic factor for upper tract transitional cell carcinoma. AB - PURPOSE: Upper tract transitional cell carcinoma (UTTCC) is a relatively rare tumor. Overall 5-year disease specific survival is in the range of 16.5% to 95% depending on stage. In this study we evaluated predictors associated with disease recurrence and disease specific survival. MATERIALS AND METHODS: We report on 129 patients with a median age of 68 years who underwent nephroureterectomy for UTTCC between July 1989 and June 2002. A total of 67 patients had primary UTTCC and 62 had previous (52) or synchronous (10) transitional cell carcinoma of the bladder (BTCC). Medical records were reviewed and analyzed for possible prognostic predictors (primary tumor stage, grade, multifocality, carcinoma in situ, symptoms and signs at presentation, sex, and history of smoking). Disease specific survival and freedom from bladder recurrence were assessed with the Kaplan-Meier method, and differences between the groups were compared using the log rank test. The Cox proportional hazards regression model was used to assess the significance of each predictor. RESULTS: Disease specific death was reported for 44 patients. In a multivariate analysis using previous BTCC as a predictor (categorized as superficial, invasive or none), primary disease stage and history of BTCC were associated with disease specific survival (p = 0.001 and p = 0.018). History of BTCC grouped the patients into distinct populations in terms of disease specific survival and freedom from bladder recurrence. CONCLUSIONS: This study demonstrates that a history of BTCC (invasive or superficial) has an adverse effect on the prognosis of patients diagnosed with UTTCC independent of primary tumor stage. PMID- 15538227 TI - Usefulness of CA 125 as a preoperative prognostic marker for transitional cell carcinoma of the bladder. AB - PURPOSE: We evaluated the role of CA 125 as a prognostic marker in patients with transitional cell carcinoma (TCC) of the bladder undergoing uniform radical cystectomy with bilateral pelvic iliac lymphadenectomy. MATERIALS AND METHODS: From April 1996 to March 2001, 287 patients with a median age of 68 years (range 34 to 89) had preoperative CA 125 serum assays performed. All patients had high grade or invasive TCC and were scheduled to undergo radical cystectomy. Preoperative CA 125 was evaluated according to tumor grade and pathological subgroups (organ confined, extravesical and lymph node positive) as well as recurrence-free and overall survival. RESULTS: Of the 287 patients 31 (10.8%) demonstrated increased serum CA 125 (greater than 35 U/ml). No significant association was found between increased CA 125 and recurrence-free survival or tumor grade. However, a statistically significant association was found among pathological tumor subgroups, namely 4.9% for organ confined, 11.3% for extravesical and 21.6% for lymph node positive disease (p <0.001). Furthermore, patients with lymph node positive disease or high grade tumors (grades III/IV and IV/IV) and increased CA 125 assays demonstrated significantly worse overall survival (p <0.001). CONCLUSIONS: Increased CA 125 was seen in approximately 11% of patients with high grade or invasive TCC preoperatively. It was more commonly found in patients with locally advanced and lymph node positive disease, and it was associated with overall survival. However, recurrence-free survival was not associated with CA 125. Further studies are required to define the exact role of CA 125 in bladder cancer. PMID- 15538228 TI - Squamous cell carcinoma arising in an augmented bladder in a patient with bladder exstrophy. PMID- 15538229 TI - Association of hemospermia with prostate cancer. AB - PURPOSE: Hemospermia is uncommon clinical condition that usually follows a benign course. The association between hemospermia and prostate cancer has been reported but to our knowledge not thoroughly investigated. We studied the incidence of hemospermia and the association between prostate cancer and hemospermia in a large prostate cancer screening population. MATERIALS AND METHODS: Between 1991 and 2001, 26,126 ambulatory men 50 years or older (40 years or older with a family history of prostate cancer or black race) underwent a community based prostate cancer screening study using serum prostate specific antigen (PSA) and digital rectal examination (DRE). PSA measurement and DRE were repeated at 6 month or 1-year intervals depending on PSA for the remainder of the study. Men underwent prostate biopsy due to increased serum PSA (greater than 4.0 ng/ml until May 1995 or greater than 2.5 ng/ml after May 1995) or suspicious DRE. Men with a history of prostate cancer were excluded from study. Men completed a questionnaire, including information about hemospermia, at each screening visit. Hemospermia information from the initial questionnaire was analyzed. The relative risk of prostate cancer diagnosis in the overall prostate cancer screening population and the cohort with hemospermia was determined. Detailed prostate cancer characteristics were evaluated in those who had hemospermia and underwent radical prostatectomy. We used a multivariate logistic regression model to test the independent significance of hemospermia after adjusting for other known predictors of prostate cancer detection. RESULTS: Prostate cancer was detected in 1,708 of the 26,126 men (6.5%) who underwent prostate cancer screening. Prostate cancer was diagnosed in 19 of the 139 men (13.7%) who reported hemospermia upon entering the prostate cancer screening study. The median age of the 139 men was 61 years (range 40 to 89). Ten of the 13 men who underwent radical retropubic prostatectomy had stage pT2 disease, while 3 had stage pT3 disease. In the logistic regression model hemospermia was a significant predictor of prostate cancer diagnosis after adjusting for age, PSA and DRE results (OR 1.73, p = 0.054). CONCLUSIONS: Hemospermia is rare (0.5%) in a prostate cancer screening population. When a man presents with hemospermia, prostate cancer screening should be vigilantly performed since hemospermia is associated with an increased risk of prostate cancer. PMID- 15538230 TI - Prostate cancer characteristics and prostate specific antigen changes in screening detected patients initially treated with a watchful waiting policy. AB - PURPOSE: We evaluated prostate cancer (PCa) characteristics at diagnosis and changes in prostatic specific antigen (PSA) with time in males with screening detected PCa that was initially managed with a watchful waiting policy. MATERIALS AND METHODS: Patients with histologically proven PCa and PSA less than 10 ng/ml were selected from the European Randomized Study of Screening for Prostate Cancer, section Rotterdam. The choice of initiating a watchful waiting policy was patient desire or physician advice. PSA slope and PSA doubling time (PSADT) were calculated in patients with 3 or more PSA tests results available. RESULTS: A total of 191 patients were included. Mean age at diagnosis was 69 years and mean PSA was 3.9 ng/ml. Of the patients 92.6% had a Gleason score of 3 + 3 or lower, 133 had a followup of greater than 12 months (mean 40) and 35 (29.2%) had a negative PSA slope. Mean PSADT was 9.7 years (range 0.3 to 155) in 85 males with a positive PSA slope. During followup 30 patients changed therapy. CONCLUSIONS: Watchful waiting remains a controversial prostate cancer treatment strategy. In select screening detected patients with PCa there appears to be a subgroup with stable or even decreasing PSA values with time. These males could profit from a watchful waiting policy with possible deferred treatment. Together with conventional tumor parameters at diagnosis PSADT and PSA slope during followup could be used to monitor tumor activity and possibly aid in determining the time of deferred treatment. Further followup is mandatory to validate these results. PMID- 15538231 TI - Ferromagnetic thermal ablation of locally recurrent prostate cancer: prostate specific antigen results and immediate/intermediate morbidities. AB - PURPOSE: Curative options for locally recurrent prostate cancer following external beam radiotherapy are limited due to the significant morbidity associated with surgical therapy. ThermoRods (Ablation Technologies, San Diego, California) are permanently implantable, 14 mm cobalt-palladium alloy rods that produce heat through oscillation of a magnetic field. The rod is designed to self regulate the temperature to 70C by a temperature dependent magnetic transition (Curie effect). We determined whether patients with prostate cancer and local failure could be treated with thermal ablation of the prostate using this novel technology. MATERIALS AND METHODS: A total of 14 men with an average age of 72 years (range 62 to 81) were enrolled in the study. All had biopsy proven prostate cancer with increasing prostate specific antigen (PSA) (1.0 to 10.3 ng/ml). The seminal vesicles were not routinely biopsied. Metastatic disease was assayed in all men with bone scan and in later patients with abdominopelvic computerized tomography. Patients had ThermoRods placed under transrectal ultrasound guidance, similar to brachytherapy. The pre-plan was rigorously followed to produce a 3 dimensional array with rods separated by 1 cm across the short axis. Patients were treated in a magnetic field for 1 hour. Urethral and rectal temperatures were also monitored and cooled appropriately. Serial PSA measurements and 6 month posttreatment biopsies were obtained after the procedure. RESULTS: Average time since radiation was 4.5 years. PSA nadir values after radiation were between 0.3 and 2.2 ng/ml. Prostatic temperatures were homogeneously increased greater than 50C, while rectal and urethral temperatures did not exceed 44C at any point. The urethral catheter was removed 2 weeks postoperatively in all cases. Six months after the procedure 8 of the 14 men (57%) had a PSA decrease to less than 0.1 ng/ml. Complications included urinary retention as well as incontinence. Incontinence was generally temporary and only 1 patient (7%) had long-term incontinence. Most patients were impotent after treatment. Of the patients 21% experienced urinary retention. Five patients (36%) needed to undergo secondary procedures after therapy, including transurethral prostate resection in 2, balloon dilation in 1, cystoscopic removal of necrotic tissue in 1 and bladder irrigation in 1. CONCLUSIONS: This novel means of thermal ablation produces significant PSA decreases in patients. Morbidity is predominantly in the form of incontinence and impotence. Incontinence is generally temporary. The use of ferromagnetic rods enables the precise control of tissue destruction with the sparing of immediately adjacent structures. The technology uses skills familiar to the urologist and avoids radiation. The durability of the PSA response and incidence of late morbidity remain to be defined. PMID- 15538232 TI - Transdermal estradiol improves bone density when used as single agent therapy for prostate cancer. AB - PURPOSE: Current androgen deprivation therapies for men with prostate cancer cause accelerated osteoporosis and a significant risk of osteoporotic fracture. We have recently shown that transdermal estradiol is an effective alternative for such patients. Here we report the impact of transdermal estradiol therapy on the bone mineral density of men with prostate cancer. MATERIALS AND METHODS: A total of 20 patients with newly diagnosed locally advanced or metastatic prostate cancer were treated with transdermal estradiol patches. Bone mineral density of the lumbar spine and the proximal femur was measured with dual-energy x-ray absorptiometry, and correlated with computerized tomography and isotope bone scan findings at 6-month intervals. RESULTS: In all measured regions bone mineral density increased with time. By 1 year mean bone mineral density +/- SEM had increased by 3.60% +/- 1.6% in the lumbar spine (p = 0.055), 2.19% +/- 1.03% in the femoral neck (p = 0.055), 3.76% +/- 1.35% in the Ward's region (p = 0.008) and 1.90% +/- 0.85% in the total hip (p = 0.031), respectively. Of 12 osteoporotic sites 4 had improvement based on World Health Organization grading. All other sites improved toward a better classification. CONCLUSIONS: Transdermal estradiol protects against bone loss in men with prostate cancer and may improve bone density in those at risk for osteoporotic fracture. PMID- 15538233 TI - Hip fractures in men with prostate cancer treated with orchiectomy. AB - PURPOSE: Androgen deprivation therapy increases the risk of osteoporosis related fractures. This issue is of increasing importance in men with prostate cancer as increasingly more undergo androgen deprivation therapy and therapy is administered sooner following diagnosis. Data directly addressing the long-term fracture risk in men diagnosed with prostate cancer are limited. MATERIALS AND METHODS: Using population based registries in Sweden we studied the incidence of hip fractures in 17,731 men diagnosed with prostate cancer from 1964 to 1996 who were treated with bilateral orchiectomy within 6 months of diagnosis. The fracture incidence was compared to the incidence in 43,230 men diagnosed with prostate cancer but not treated with orchiectomy and in 362,354 of similar age who were randomly selected from the general population. RESULTS: Men treated with orchiectomy were at increased risk for hip fracture. The estimated relative risk comparing men who underwent orchiectomy to population controls was 2.11 (95% CI 1.94 to 2.29) for femoral neck fractures and 2.16 (95% CI 1.97 to 2.36) for intertrochanter fractures. An increased risk of hip fracture was observed as early as 6 months after orchiectomy and the relative risk remained fairly constant up to 15 years following orchiectomy. CONCLUSIONS: Hip fracture risk increases almost immediately following orchiectomy and the excess risk persists for at least 15 years. This side effect should be considered when assessing the merits of androgen deprivation therapy, particularly in symptom-free men diagnosed with localized prostate cancer. Measures to prevent osteoporosis should be considered in men undergoing androgen deprivation therapy. PMID- 15538234 TI - Prognostic value of anemia in newly diagnosed metastatic prostate cancer: a multivariate analysis of southwest oncology group study 8894. AB - PURPOSE: Previously reported association of anemia with shorter survival in newly diagnosed metastatic prostate cancer may simply reflect extent of disease. The impact of anemia on response to androgen deprivation is not known. We examined the prognostic value of anemia in a multivariate analysis that included disease extent and other tumor and demographic covariates in 957 patients starting hormonal therapy for metastatic prostate cancer as part of Southwest Oncology Group study 8894. MATERIALS AND METHODS: The multivariate associations of disease and patient measures with anemia (hemoglobin less than 12 gm/dl) were evaluated with a logistic regression model. The associations between hemoglobin and survival and progression-free survival (PFS) were evaluated using a proportional hazards model, and included indicators for quartiles of hemoglobin and baseline covariates. Prostate specific antigen (PSA) normalization (PSAN, or PSA of 4 ng/ml or less) was evaluated with a logistic model. RESULTS: Quartiles of hemoglobin were 10.1 or less, 10.2 to 12.0, 12.1 to 13.4 and greater than 13.4 gm/dl. In a multivariate model anemia was significantly associated (p <0.02) with being black, performance status 2 to 3 (vs 0 to 1), extensive disease and higher PSA. Anemia was inversely associated with prior therapy with curative intent and with Gleason score 6 to 7 (vs 5 or less), and was not associated with age or bone pain. After adjusting for potential confounders, lower hemoglobin was associated with shorter survival and PFS, and lower likelihood of PSAN with hormonal therapy (p <0.01). CONCLUSIONS: In this newly diagnosed metastatic prostate cancer sample, anemia was common and was associated with shorter survival, shorter PFS, and lower likelihood of PSAN with hormonal therapy after adjustment for disease status and other covariates. PMID- 15538235 TI - Transperitoneal or extraperitoneal laparoscopic radical prostatectomy: does the approach matter? AB - PURPOSE: : The greater accuracy of apical dissection and reconstruction in our first 100 patients undergoing transperitoneal laparoscopic radical prostatectomy (TLRP) was not matched by a proportionate increase in the rate of return to normal continence compared with our prior open prostatectomy experience. We postulated that greater bladder dysfunction due to the almost total bladder dissection mandated by TLRP might be responsible and this might be rectified by the adoption of laparoscopic radical prostatectomy using an extraperitoneal approach (ELRP). MATERIALS AND METHODS: : A total of 100 patients undergoing TLRP were compared with 100 undergoing ELRP. The groups were subdivided into halves to investigate the influence of any learning curve effect. All patients had clinical stage T3aN0M0 or less prostate cancer and they were operated on by a single surgeon. RESULTS: : Mean operative time (238.9 vs 190.6 minutes), blood loss (310.5 vs 201.5 ml), postoperative hospitalization (3.8 vs 2.6 nights) and catheterization duration (11.3 vs 10.1 days) were significantly greater in the TLRP group. After the first 50 cases were excluded in each group statistical significance persisted only for operative time (218.3 vs 184.2 minutes) and hospitalization (3.5 vs 2.5 nights). The pad-free rate was significantly lower 3 months following ELRP (80% vs 56%, p = 0.02). The overall 12-month pad-free rate for TLRP and ELRP was 90% and 96%, respectively. The overall 12-month erection rate for TLRP and ELRP was 61% and 82%, respectively. CONCLUSIONS: : ELRP is superior to TLRP with respect to operative time, hospitalization and early continence. PMID- 15538236 TI - Increased accuracy of biopsy Gleason score obtained by extended needle biopsy. AB - PURPOSE: Accurate tumor grading is critical for adequate prostate cancer treatment. Nonetheless, the Gleason score of standard sextant biopsy correctly predicts the Gleason score of the radical prostatectomy specimen in about 50% of cases. We investigated if extended needle biopsy could improve biopsy Gleason score accuracy. MATERIALS AND METHODS: Laparoscopic transperitoneal radical prostatectomy was performed in 135 patients. Prostate cancer was diagnosed in 89 cases by standard sextant transrectal (6 to 8 cores) biopsy and in 46 by extended needle (12 core transperineal under transrectal guidance) biopsy. Preoperative evaluation included digital rectal examination, prostatic specific antigen measurement, transrectal ultrasonography and endorectal coil magnetic resonance imaging in all patients. All biopsy and prostatectomy specimens were reviewed by a single pathologist. RESULTS: Clinical characteristics were similar in the 2 groups. The concordance between prostate biopsy and radical prostatectomy Gleason score was 32 of 46 cases (70%) and 44 of 89 (49%) for 12 core and standard transrectal biopsy, respectively (z test p = 0.0127). Biopsy under grading was found in 11 of 46 cases (24%) and 35 of 89 (39%) (z test p = 0.0366), and biopsy over grading was found in 3 of 46 (6%) and 10 of 89 (11%) (z test p = 0.1894) with 12 core and standard transrectal biopsy, respectively. Primary Gleason pattern was predicted exactly by biopsy in 40 of 46 cases (87%) and 56 of 89 (63%) with 12 core and standard sextant biopsy, respectively (z test p = 0.0018). CONCLUSIONS: Extended needle biopsy significantly increases the accuracy of biopsy Gleason score for assessing final prostate cancer grade. PMID- 15538237 TI - Potency, continence and complications in 3,477 consecutive radical retropubic prostatectomies. AB - PURPOSE: We report results in a series of 3,477 consecutive patients treated with anatomical nerve sparing radical retropubic prostatectomy (RRP) in terms of recovery of erectile function, urinary continence and postoperative complications. MATERIALS AND METHODS: From May 1983 through February 2003, 1 surgeon (WJC) performed anatomical RRP using a unilateral or bilateral nerve sparing modification when possible. Urinary continence and recovery of erections were evaluated in men with a minimum followup of 18 months. Excluded from potency analysis were men who were not reliably potent before surgery, those who did not undergo a nerve sparing procedure and those who received postoperative adjuvant radiotherapy or hormonal therapy within 18 months of surgery. Other postoperative complications in this patient population were also evaluated. RESULTS: Erections sufficient for intercourse occurred in 76% of preoperatively potent men treated with bilateral (1,770) and 53% of men treated with unilateral or partial nerve sparing (64) surgery. Adequate erectile function was more common following bilateral than unilateral nerve sparing surgery in men younger than 70 years old (78% versus 53%, p = 0.001) compared with those 70 years old or older (52% versus 56%, p = 0.6). Recovery of urinary continence occurred in 93% of all men and was associated with younger age (p = 0.001) but not nerve sparing surgery, tumor stage, prostate specific antigen (PSA), Gleason grade or number of prior prostatectomies performed by the surgeon. Postoperative complications occurred in 320 (9%) of patients and were associated with older age (p <0.0001), nonnerve sparing surgery (p = 0.001), PSA era (p <0.0001) and surgeon experience. Complications were not significantly correlated with clinical stage, pathological stage, preoperative PSA or Gleason grade. There was no perioperative mortality. CONCLUSIONS: Nerve sparing RRP can be performed with favorable potency and urinary continence. Better results are achieved in younger men. Other complications are reduced with increasing surgeon experience. PMID- 15538238 TI - 10-year disease-free survival rates after simultaneous irradiation for prostate cancer with a focus on calculation methodology. AB - PURPOSE: We documented the 10-year disease-free survival rate after simultaneous irradiation for prostate cancer and suggested standards for outcome calculation methodology. MATERIALS AND METHODS: From 1992 to 1998, 1,469 consecutive men with clinical stage T1T2NxM0 prostate cancer who did not receive neoadjuvant hormones were treated with simultaneous irradiation, an ultrasound guided transperineal prostate I seed implant followed by external irradiation. Median pretreatment prostate specific antigen (PSA) was 7.1 ng/ml (range 0.3 to 88). All men were treated 5 or more years ago. Median followup was 6 years (range 3 months to 11 years). Disease freedom was defined as the achievement and maintenance of PSA 0.2 ng/ml or less, and treatment failure was defined as a PSA nadir greater than 0.2 ng/ml or a subsequent PSA increase above this level. RESULTS: The overall 10-year disease-free survival rate was 83%. Median time to recurrence was 30 months (range 3 months to 8 years) and 24% of recurrences were after 5-year followup. The 10-year outcome according to low, intermediate and high risk group was 93%, 80% and 61%, respectively (p <0.0001). Multivariate analysis of factors related to disease freedom documents that pretreatment PSA, Gleason score and percent positive biopsies were significant but stage and age were not. CONCLUSIONS: By calculating outcome with PSA cut point 0.2 ng/ml and evaluation only of men treated 5 or more years ago, the 10-year disease-free survival rates from this study can be reasonably compared with the outcome of radical prostatectomy performed in the PSA era. PMID- 15538239 TI - Morbidity and functional outcomes of salvage radical prostatectomy for locally recurrent prostate cancer after radiation therapy. AB - PURPOSE: Few patients with locally recurrent prostate cancer after external beam (EB) or interstitial (I) radiotherapy (RT) are considered candidates for salvage radical prostatectomy (RP) due to high reported rates of major complications and urinary incontinence. We report the morbidity associated with salvage RP in 100 consecutive patients. MATERIALS AND METHODS: From 1984 to 2003 salvage RP was performed for recurrent prostate cancer following EBRT in 58 cases, and IRT in 42 (28 retropubic) and (14 transperineal). Clinical information was obtained from a prospective database. RESULTS: Since 1993, the major complication rate has decreased significantly (13% vs 33%, p = 0.02), including the rectal injury rate (2% vs 15%, p = 0.01). Compared with retropubic IRT and/or pre-radiotherapy pelvic lymph node dissection the risk of major complications following EBRT or transperineal IRT was significantly less (OR 0.2, p = 0.006). At 5 years an estimated 39% of patients were dry and 68% required 1 pad daily or less. A total of 23 patients with moderate-severe incontinence underwent artificial sphincter placement. The anastomotic stricture rate was 30%. The 5-year potency rate was 28% following unilateral or bilateral nerve sparing RP and 45% in previously potent patients. CONCLUSIONS: Due to patient selection the major complication rate after salvage RP has improved significantly with time and it is similar to that of standard RP. Rates of anastomotic stricture and moderate to severe incontinence are higher than those observed after standard RP. However, most patients recover reasonable urinary continence and a substantial number of select patients recover potency. The acceptable morbidity profile of salvage RP following EBRT and transperineal IRT should persuade more physicians to consider patients for this potentially curative procedure. PMID- 15538240 TI - Prostate specific antigen doubling time subsequent to radical prostatectomy as a prognosticator of outcome following salvage radiotherapy. AB - PURPOSE: Therapy for men with detectable prostate specific antigen (PSA) following radical prostatectomy (RP) for prostate cancer remains controversial. Salvage radiotherapy (SRT) is commonly used because of its relatively low morbidity. We present a single institution retrospective review of patients treated with SRT. MATERIALS AND METHODS: A longitudinal cohort study (between April 1987 and April 2000) using the referral based Mayo Clinic Prostate Cancer Registry was conducted. A total of 211 patients were included in this study if detectable serum PSA was the sole indication for SRT and no hormonal therapy was administered. RESULTS: Median followup from surgery to death or last followup was 7.2 years, from RP to SRT was 1.7 years and from SRT to last contact was 4.2 years. Median PSA and prostate specific antigen doubling time (PSADT) at SRT initiation was 0.60 ng/ml and 7.32 months; respectively. Of the patients 90% had nadir PSA less than 0.4 ng/ml within 3 years of SRT. Biochemical disease-free rates at 5 years for PSADT less than 12 or 12 months or greater was 48% and 66%; respectively (p = 0.080). By 10 years there was no significant difference in biochemical disease-free rate (34% vs 35%). Clinical metastasis (10% and 29%) developed in patients with a PSADT less than 12 months at a significantly higher rate than in patients with a PSADT of 12 months or more (0% and 17%, p = 0.045) at 5 and 10 years, respectively. Multivariate analysis revealed pre-SRT PSADT (less than 12 months, H.R. 3.88, p = 0.032), seminal vesicle invasion (H.R. 3.22, p = 0.008), pathological grade (H.R. 1.58, p = 0.023) and PSA at SRT (H.R. 1.29 for a 2-fold increase, p = 0.044) to be significant independent predictors of clinical recurrence. The interval from RP to SRT did not add to the model (p = 0.22). CONCLUSIONS: A biochemical response can be expected in the majority of patients within 3 years of receiving SRT. Patients with a pre-SRT PSADT of 1 year or less have a less sustained biochemical response to SRT than patients with a PSADT greater than 1, yet the majority of patients appear to receive long-term benefit from this adjunctive therapy. PSADT is an independent predictor of biochemical and clinical disease recurrence following SRT. PMID- 15538241 TI - Perineural invasion in radical prostatectomy specimens: lack of prognostic significance. AB - PURPOSE: The prognostic significance of perineural invasion in radical prostatectomy specimens is uncertain. We evaluated the relationship between perineural invasion and other pathological characteristics in whole mount radical retropubic prostatectomy specimens as well as prostate specific antigen (PSA) recurrence postoperatively. MATERIALS AND METHODS: Between 1999 and 2003, 364 consecutive patients were treated with radical prostatectomy for localized prostate cancer. Radical prostatectomy specimens were processed by the whole mount technique. The relationship of perineural invasion and various clinicopathological characteristics to PSA recurrence was analyzed. RESULTS: Perineural invasion was present in 287 specimens (79%). Specimens with perineural invasion were associated with smaller prostate weight (p <0.0001), greater pathological stage (p <0.0001), larger tumor volume (p <0.0001), higher Gleason score (p <0.0001), a higher incidence of extraprostatic extension (p <0.0001) and seminal vesicle invasion (p = 0.02), and a higher positive surgical margin rate (p = 0.01). Perineural invasion did not correlate with preoperative PSA (p = 0.96), lymph node metastases (p = 0.35), multifocality (p = 0.21), high grade prostatic intraepithelial neoplasia (p = 0.12) or PSA recurrence (p = 0.24). CONCLUSIONS: While perineural invasion in the radical prostatectomy specimen significantly correlated with multiple adverse pathological factors, it did not predict which patients will have early PSA recurrence following radical prostatectomy. PMID- 15538242 TI - Prognosis of patients with lymph node positive prostate cancer following radical prostatectomy: long-term results. AB - PURPOSE: We determined the prognostic factors that affect recurrence and survival in patients with lymph node positive prostate cancer. MATERIALS AND METHODS: Between 1972 and 1999, 1,936 patients underwent radical retropubic prostatectomy and pelvic lymph node dissection for clinically organ confined prostate cancer. A total of 235 patients (12.1%) were found to have disease metastatic to the lymph nodes (stage D1). Of the patients 69% received no adjuvant treatment. We reviewed the tumor stage (TNM), Gleason score, number and percent of involved lymph nodes (lymph node density), preoperative prostate specific antigen when available and adjuvant treatment. Overall survival and recurrence-free survival were estimated using Kaplan-Meier plots. RESULTS: Followup was 1 to 24 years (median 11.4). Overall median survival was 15 years. Overall clinical recurrence-free survival at 5, 10 and 15 years was 80%, 65% and 58%, respectively. Patients who had 1 or 2 positive lymph nodes had a clinical recurrence-free survival of 70% and 73% at 10 years, respectively, vs 49% in those who had 5 or more involved lymph nodes (p = 0.0031). When stratified by lymph node density, patients with a lymph node density of 20% or greater were at higher risk for clinical recurrence compared to those with a density of less than 20% (relative risk = 2.32, p <0.0001). On stratified log rank test only prostate cancer T stage, and the number and percent of positive lymph nodes correlated with recurrence-free and overall survival. CONCLUSIONS: Local tumor bulk and the number/percent of involved lymph nodes significantly affect disease progression and the survival rate. Radical prostatectomy may offer long-term survival in patients who have limited tumor bulk and nodal involvement. PMID- 15538243 TI - Is urine methylhistamine a useful marker for interstitial cystitis? AB - PURPOSE: We determined whether urine methylhistamine (MH) is associated with other clinical features and bladder biopsy findings in interstitial cystitis (IC). MATERIALS AND METHODS: Urine MH and University of Wisconsin symptom scores were measured before and 1 month after bladder distention for IC. Bladder biopsies were done at the time of distention. In new patients with IC who met cystoscopic criteria urine MH was compared before and after distention, and changes in urine MH were compared with changes in symptom scores. Pre-distention urine MH was tested for associations with the results of a symptom questionnaire, symptom response after distention, cystoscopic findings and bladder biopsy findings. RESULTS: There were no significant associations between urine MH and symptom scores, response to bladder distention, cystoscopic findings or bladder biopsy features, including mast cell count by tryptase staining. Urine MH was similar in new patients with IC who did vs did not meet cystoscopic criteria. Urine MH was similar in new vs chronically treated patients with IC. CONCLUSIONS: Urine MH is unlikely to be useful as an objective marker of the response to bladder distention, as a method to predict which patients will respond to bladder distention or as a noninvasive substitute for bladder biopsy and mast cell counts by tryptase staining. PMID- 15538244 TI - Nitric oxide as an objective marker for evaluation of treatment response in patients with classic interstitial cystitis. AB - Nitric oxide (NO) has previously been shown to be a marker for inflammatory disorders in the bladder. We investigated if the measurement of NO can be used to evaluate the treatment response in classic interstitial cystitis (IC). MATERIALS AND METHODS: A total of 15 patients diagnosed with classic IC were included. The patients were treated with oral prednisolone for 8 weeks. We evaluated patient symptoms/problems with the IC index, and we measured NO formation in the bladder. RESULTS: Seven patients were classified as responders with a 4 point or more decrease in symptom score after 8 weeks of treatment. In this group the mean symptom score was +/- SEM decreased from 15 +/- 1 to 7 +/- 2 at the end of therapy (p <0.05). The problem score was also significantly reduced. Responders showed a clear decrease in bladder NO. The 8 nonresponders did not show any improvement in symptom/problem score and there was no change in bladder NO during steroid treatment. Furthermore, the study showed a statistically significant correlation between changes in symptom/problem score and changes in luminal bladder NO in each patient. CONCLUSIONS: The study shows that NO can be used not only to measure inflammation in patients with IC, but also to evaluate objectively the treatment response in individuals. This makes NO formation a useful marker in the assessment of classic interstitial cystitis. PMID- 15538245 TI - Vesicolymphocele fistula following sclerotherapy for lymphocele. PMID- 15538246 TI - Elucidation of factors determining formation of calcium phosphate stones. AB - PURPOSE: In this study we compared the effect of the urinary concentration of calcium with that of oxalate on the urinary relative saturation ratio (RSR) of calcium oxalate and brushite in patients with pure or mixed calcium phosphate stones. MATERIALS AND METHODS: A retrospective data analysis was performed of urinary stone risk analysis in 133 patients with pure or mixed calcium phosphate stones (30% or greater). Correlation was sought between stone events and urinary stone risk factors, and between brushite and urinary constituent RSRs before and after adjustment for variables. RESULTS: The formation rates of total events and stone surgeries weakly but significantly correlated with RSR urinary brushite (total events r = 0.22, p = 0.01) but not with RSR calcium oxalate. Urinary RSR brushite significantly correlated positively with urinary calcium (r = 0.67, p <0.0001), phosphorus and pH. After adjustment RSR brushite inversely correlated with citrate (r = -0.3, p = 0.0006). However, it did not correlate with oxalate. CONCLUSIONS: In stone formers with calcium phosphate or mixed calcium oxalate calcium phosphate stones the stone formation rate depends on the urinary saturation of brushite but not of calcium oxalate. RSR brushite is determined by the urinary concentration of calcium, phosphate, citrate and pH but not by oxalate. Hypercalciuria should be controlled in patients with idiopathic calcareous renal calculi who form stones of pure or mixed calcium phosphate. PMID- 15538247 TI - A multivariate analysis of risk factors associated with subcapsular hematoma formation following electromagnetic shock wave lithotripsy. AB - PURPOSE: Subcapsular or perinephric hematoma is one of the most frequent and potentially serious complications of extracorporeal shock wave lithotripsy (SWL). We determined the incidence of and risk factors for renal hematomas following electromagnetic shock wave lithotripsy. MATERIALS AND METHODS: Between February 1999 and August 2003, 570 SWL treatments were performed using a Modulith SLX electromagnetic lithotriptor (Storz, St. Louis, Missouri). A total of 415 of these treatments in 317 patients were performed for stones in the renal pelvis or calices and these treatment episodes represent the study group reported. Treatment episodes were reviewed from a prospective institutional review board approved registry and analyzed for patient age, gender, body mass index, mean arterial pressure at induction, stone location, total number of shock waves and peak shock wave intensity. RESULTS: Following these 415 episodes subcapsular or perinephric hematomas developed in 17 patients for an overall incidence of 4.1%. The probability of hematoma after shock wave lithotripsy increased significantly as patient age at treatment increased, such that the probability of hematoma was estimated to be 1.67 times greater for each 10-year incremental increase in patient age. None of the other variables analyzed were significantly related to the incidence of hematoma formation at the 0.05 level. CONCLUSIONS: The incidence of renal hematoma formation following electromagnetic SWL for renal calculus was 4.1%. The probability of hematoma increased significantly with increasing patient age but it was not associated with increasing mean arterial pressure at treatment. These findings are in contrast to previous reports of hematoma associated with electrohydraulic SWL. These differences may be a consequence of the smaller focal zone and higher peak pressure associated with Storz Modulith electromagnetic SWL and, just as importantly, a consequence of the difference in the manner in which blood pressure was defined. PMID- 15538248 TI - Cost-effectiveness of medical management strategies for nephrolithiasis. AB - PURPOSE: We evaluated the cost-effectiveness and stone recurrence rates of common management strategies in stone formers. MATERIALS AND METHODS: A decision tree model was created to compare the costs of 6 medical treatment strategies, namely dietary measures alone (conservative), empiric drug treatment (empiric), or directed drug therapy based on simple or comprehensive metabolic evaluation. The model assumed cost accrual for evaluation, medications, emergency treatment and surgery for stone recurrence. We determined recurrence rates, medical evaluation sensitivity and the risk reduction of medical treatments from the literature and the costs of surgery, emergency room and medical evaluation at our county hospital. Drug costs were obtained from 2 national pharmacy chains. RESULTS: In first time stone formers conservative treatment was the least costly strategy and it yielded a stone formation rate of 0.07 stones per patient yearly. In recurrent stone formers conservative treatment was less costly than drug treatments but it was associated with a higher stone recurrence rate (0.3 stones per patient yearly). The remaining drug treatments were more costly than conservative treatment ($885 to 1,187 vs $258 yearly) but they further decreased recurrence rates by 60% to 86%. Modified simple medical evaluation and management (SMEM), that is drug treatment in all patients after simple evaluation) was slightly more costly than empiric treatment and minimally more effective. Comprehensive evaluation and treatment offered no advantage in cost or efficacy over empiric treatment or modified SMEM. CONCLUSIONS: In first time stone formers conservative therapy is cost-effective and efficacious. In recurrent stone formers conservative therapy is unsatisfactory despite low cost because of a high recurrence rate. Modified SMEM and empiric therapy yield the same cost and efficacy as treatments that follow comprehensive evaluation. However, modified SMEM may be advantageous because it offers useful additional medical information. PMID- 15538249 TI - Survey of urological laparoscopic practice patterns in the midwest. AB - PURPOSE: Laparoscopic surgery is assuming a greater role in the management of many urological disorders. We performed a survey of urologists in the Midwest United States to define laparoscopic practice patterns regarding urological disease. MATERIALS AND METHODS: Surveys were mailed to 1,450 members of the North Central Section of the American Urological Association. Questions pertained to age, practice demographics, residency training and amount of laparoscopy performed. RESULTS: A response rate of 33% was obtained. A total of 49% of urologists perform no laparoscopy, 30% devote less than 5% of their practice and 21% devote 5% or more of their practice to laparoscopy. Of the responders 15% believed they were adequately trained during residency to perform laparoscopy. Among those who performed hand assisted laparoscopy (HAL), 52% used it to gain familiarity with laparoscopic techniques. There was an inverse correlation between time in practice and amount of laparoscopy performed (p <0.0001). Urologists in academia perform laparoscopy at a greater proportion versus those in private practice (p <0.01). Urologists who incorporated laparoscopy during fellowship performed the most laparoscopy (p = 0.003). Those who considered themselves adequately trained during residency were more likely to perform laparoscopy (p <0.0001). Urologists who perform HAL devote more of their operating time to laparoscopy than those who do not (p <0.0001). The number of laparoscopic cases performed correlated with decreased time in practice (p <0.0001), academic setting (p = 0.001) and use of HAL (p <0.0001). CONCLUSIONS: Urological laparoscopic surgery remains at an early stage of development in the Midwestern United States. Although HAL has widened the scope of laparoscopy among urologists, increased training for urologists and residents is necessary. PMID- 15538250 TI - Differences in complications and outcomes for obese patients undergoing laparoscopic radical, partial or simple nephrectomy. AB - PURPOSE: Obesity has increased dramatically in American society during the last 2 decades. While the laparoscopic approach is common for patients requiring radical and partial nephrectomy, it is unclear if this procedure leads to worse outcomes and complications in obese patients. We determined if obese patients undergoing laparoscopic radical (RN), partial (PN) and simple (SN) nephrectomy are at risk for worse surgical outcomes or increased complications. MATERIALS AND METHODS: We retrospectively identified patients treated with nontransplant transperitoneal laparoscopic nephrectomies from 1998 to 2003. Patients with missing body mass index (BMI), operative, postoperative or pathological information were excluded from study. Obese patients (BMI 30 or greater) were compared to nonobese patients (BMI less than 30). RESULTS: A total of 189 patients undergoing 117 RN, 44 PN and 30 SNs met study criteria, and 29.0% of patients were obese. Overall obese patients had longer operative times (280 versus 241 minutes, p = 0.003), greater estimated surgical blood loss (230 versus 109 ml, p = 0.0001) and higher transfusion rates (6.8% versus 0.8%, p = 0.032) than nonobese patients. In subgroup analyses obese patients receiving RN and PN had longer operative times and increased blood loss. Obese and nonobese patients have similar open conversion rates, analgesic requirements, hospital stay, time to oral intake, and major and minor complication rates regardless of nephrectomy type. CONCLUSIONS: Laparoscopic nephrectomy is associated with slightly greater operative time, estimated blood loss and transfusion rates in obese patients. Laparoscopic RN, PN and SN are safe and well tolerated in obese patients. Obesity is not a contraindication to laparoscopic renal surgery. PMID- 15538251 TI - Eliminating knot tying during warm ischemia time for laparoscopic partial nephrectomy. AB - PURPOSE: Rapid intracorporeal suturing represents a challenge when performing laparoscopic partial nephrectomy (LPN). During warm ischemia time (WIT) knot tying is a major time-consuming step. We present our technique of eliminating knot tying during LPN and the outcomes of our initial series. MATERIALS AND METHODS: Between October 2002 and October 2003, 32 patients underwent LPN for clinical T1a renal tumors. Our technique includes initial placement of a 5Fr ureteral catheter for collecting system irrigation. The renal hilum is clamped, the tumor is sharply excised and freehand suturing of the collecting system and renal parenchyma is performed using 2-zero and zero polyglactin sutures prepared with an absorbable clip (LapraTy, Ethicon Endosurgery Inc., Piscataway, New Jersey) at the terminal end. Once the suture is passed an additional clip is used to cinch it down, obviating the need for knot tying. This technique is used for closure of the collecting system as well as for placement of parenchymal compressive sutures over bolsters. Subsequent testing for watertightness with methylene blue solution is performed. RESULTS: Pathological mean tumor size was 2.1 cm (range 0.3 to 4.2). Mean operative time was 224.2 minutes (range 105 to 396). In 21 cases (65.6%) the collecting system was entered, necessitating further suturing. Mean WIT was 33.1 minutes (range 13 to 55) and mean estimated blood loss was 222.7 cc (range 5 to 600). No postoperative bleeding or urine leaks were encountered in this series. CONCLUSIONS: The use of LapraTy clips as an alternative to knot tying in LPN is safe and efficient. It simplifies the procedure and allows completion of the necessary suturing tasks during an acceptable WIT. PMID- 15538252 TI - Bladder neck closure with lower urinary tract reconstruction: technique and long term followup. AB - PURPOSE: Bladder neck closure (BNC) is an important component of reconstructive urological surgery, especially in the management of neurogenic bladder. To our knowledge we present the largest series of patients who have undergone this procedure. MATERIALS AND METHODS: A total of 39 patients with lower urinary tract (LUT) dysfunction secondary to neurogenic bladder underwent transabdominal BNC and simultaneous LUT reconstruction between 1988 and 2002. Charts were reviewed and patients were retrospectively interviewed to ascertain demographics, previous urological surgeries, perioperative data, postoperative results and complications. RESULTS: Mean postoperative followup was 36.9 months (range 7 to 173). Concomitant procedures included ileovesicostomy in 19 patients (49%), augmentation enterocystoplasty with continent cutaneous stoma in 19 (49%) and revision of a previous Mitrofanoff appendicovesicostomy in 1. The overall complication rate was 31% with a vesicourethral fistula in 6 patients (15%), of whom 4 required eventual transabdominal or transvaginal surgical correction. No other problems directly related to BNC were identified. Patients were followed by serial renal ultrasound and abdominal x-ray with upper tract maintenance seen in all patients. CONCLUSIONS: Bladder neck closure with simultaneous urinary diversion is a highly effective, well tolerated treatment for many pathological processes of the LUT. Risks for prolonged urethral leakage include high pressure systems, prior bladder neck surgery and noncompliance with catheter/drain management. These data support our belief that a high degree of success with an acceptable complication rate is attainable with careful adherence to surgical technique, proper patient selection, appropriate early postoperative management and rigid surveillance. PMID- 15538253 TI - Stomal complications of ileal conduits are significantly higher when formed in women with intractable urinary incontinence. AB - PURPOSE: The ileal conduit is held to be the safest and simplest form of urinary diversion. There are few reports about long-term problems after ileal conduit formation, especially intractable urinary incontinence in females. We reviewed long-term stomal complications in patients with an ileal conduit. MATERIALS AND METHODS: Notes on 93 consecutive patients in whom an ileal conduit was created were reviewed. Information was collected on patient demographics, indications for an ileal conduit and long-term complications, in particular parastomal and incisional hernias, stomal retraction, stenosis or prolapse and the development of a redundant loop. Mean followup available was 63.4 months (range 1 to 434). RESULTS: A total of 33 males with a mean age of 60.1 years (range 2 to 78) and 60 females with a mean age of 48.2 years (range 4 to 79) underwent ileal conduit diversion. The main indications for an ileal conduit were intractable incontinence in 44 patients (47%), cancer in 31 (33%) and interstitial cystitis in 8 (9%). In male, continent female and incontinent female patients A parastomal hernia developed in 3 (9%), 2 (9.5%) and 12 (31%), an incisional hernia developed in 1 (3%), 1 (4.8%) and 2 (5%), stomal retraction developed in 0, 2 (9.5%) and 12 (31%), stomal stenosis developed in 0 (0%), 1 (4.8%) and 6 (15.4%), and a redundant loop developed in 0 (0%), 2 (9.5%) and 5 (12.8%), respectively. A total of 23 patients (24.7%) required further surgery for stomal problems with 13 (57%) requiring more than 1 reoperation, of whom 9 were incontinent females. CONCLUSIONS: An ileal conduit is associated with a stomal complication rate of 34.4% (61% in incontinent females and 18% in other patients) and a 4.3% incisional hernia rate. Reoperation is required for stomal complications in 24.7% of cases. Stomal complication rates and reoperation rates vary by sex and the indication for ileal conduit, and they are significantly higher for those performed for intractable urinary incontinence in females. PMID- 15538254 TI - The incidence of crossing vessels in patients with normal ureteropelvic junction examined with endoluminal ultrasound. AB - PURPOSE: We estimated the incidence of a crossing vessel at the normal ureteropelvic junction (UPJ) in patients undergoing ureteroscopy and endoluminal ultrasonography for indications other than UPJ obstruction. MATERIALS AND METHODS: Endoluminal ultrasonography was performed in 141 patients undergoing upper tract endoscopy for various indications excluding UPJ obstruction. A detailed description of the anatomy of the UPJ as well as the location and size of crossing vessels was included in the operative note. Charts were reviewed to determine the precise anatomy of the UPJ. RESULTS: The overall incidence of crossing vessels at the unobstructed UPJ was 19.2%. Endoluminal ultrasonography demonstrated a crossing vessel in 13.2% of patients with ureteral narrowing or stricture, 31.3% of those with tumors or filling defects, 10.5% of those with submucosal calculi and 16.7% of patients with ureteral diverticula. In cases where the position of a crossing vessel was ascertained, 41% were anterior to the ureter, 28% anterolateral, 24% anteromedial and 7% posterior. There was a statistically significant difference in the frequency of vessels at the UPJ in patients with and without obstruction examined with endoluminal ultrasound (p <0.0001). CONCLUSIONS: A crossing vessel at the UPJ is seen with endoluminal ultrasound in 19.2% of patients with a normal UPJ. This incidence is lower than that seen in patients with obstructed UPJ. Many of these vessels are related to the lateral surface of the UPJ and there was no area that was always free of vessels. PMID- 15538255 TI - Extraperitoneal laparoscopic pyeloplasty for primary and secondary ureteropelvic junction obstruction. AB - PURPOSE: Greater experience with urological laparoscopy has lead to increasing interest in its use for reconstructive surgery, such as pyeloplasty. MATERIALS AND METHODS: A total of 124 cases of laparoscopic pyeloplasty were performed, of which 11 followed failed primary treatment done elsewhere, namely balloon dilation (3), endopyelotomy (3); open pyeloplasty (3), endopyelotomy plus balloon dilation (1) and open pyeloplasty plus balloon dilation (1). Nine patients had renal calculi. A 4 port, balloon dissecting, extraperitoneal laparoscopic approach was used in all except 1 patient, who had a horseshoe kidney, necessitating a transperitoneal approach. RESULTS: Operative time was 29 minutes longer in the secondary pyeloplasty group compared to primary cases (173.3 vs 144.0 minutes) but the conversion rate (0% vs 1.6%) and duration of postoperative hospitalization (2.8 nights each) were no greater. The complication rate was 3.6% and 9.1%, respectively. The success rate was 98.2% and 90.9% (p = 0.63) at a mean followup of 20.2 and 19.7 months, respectively. In the 9 patients with renal calculi a total of 18 calculi (94.7%) were successfully removed and the ureter was transposed medial to a crossing vessel in 50.0%. Trainee operating did not significantly prolong the procedure vs no training (162.0 vs 143.9 minutes, p = 0.06). CONCLUSIONS: Extraperitoneal laparoscopic dismembered pyeloplasty is capable of addressing all causes of ureteropelvic junction obstruction with excellent functional results and low morbidity, and with an operative time similar to that of open pyeloplasty. Secondary laparoscopic pyeloplasty does not increase hospitalization, conversion or complication rates. PMID- 15538256 TI - An automated penile compression release maneuver as a noninvasive test for diagnosis of bladder outlet obstruction. AB - PURPOSE: We tested the hypothesis that the previously described penile urethral compression release (PCR) maneuver provides a valid diagnosis of bladder outlet obstruction (BOO) using automated rather than manual penile compression by controlled inflation of a penile cuff. We also investigated urodynamic events underlying generation of the PCR index. MATERIALS AND METHODS: A total of 150 subjects attending for pressure flow studies were studied using conventional and noninvasive cystometry. Patients were classified into urodynamic diagnostic groups using standard invasive studies. The PCR index was calculated for each individual from noninvasive penile cuff data and the results were summarized for each group. ROC analysis of the PCR index was performed to define an optimum threshold for BOO diagnosis. Simultaneous invasive and noninvasive data were used to define the relationship between the PCR index, bladder contractility and the maximum flow rate. RESULTS: The mean PCR index +/- SD was significantly higher in the BOO group compared to the normal cystometry group (215% +/- 84% vs 93% +/- 39, p <0.01). ROC analysis showed that a PCR index of greater than 160% diagnosed BOO with 78% sensitivity, 84% specificity and a positive predictive value of 69%. There was a strong positive correlation between the PCR index and isovolumetric detrusor pressure, which is a measure of bladder contractility (r = 0.44, p <0.01). CONCLUSIONS: The results of this study suggest that the PCR index combines valid estimates of bladder contractility and the maximum flow rate, and it represents a clinically useful, noninvasive urodynamic parameter for the diagnosis of BOO. PMID- 15538257 TI - The effect of botulinum-A toxin on patients with severe urge urinary incontinence. AB - PURPOSE: We determined the effect of 150 units of botulinum-A toxin (Botox, Allergan, Irvine, California) on subjects with severe urge urinary incontinence (UUI). MATERIALS AND METHODS: This was an open label uncontrolled clinical trial. Subjects were recruited from the female urology and urogynecology clinics at Duke University. Inclusion criteria included evidence of UUI on 3-day bladder diary, a 24-hour pad weight of 100 gm or greater, absent or minimal stress leakage, absent detrusor dysfunction, and a history of failed anticholinergic and physical therapies. Exclusion criteria included evidence of a urinary tract infection, or other correctable or neurological etiology for UUI. The detrusor of each subject was injected with 150 units of botulinum-A toxin. Evaluations were performed at 2 weeks, 6 weeks, 3 months and 6 months after injection. Outcome measures included daily incontinence episodes, Urogenital Distress Inventory and the Incontinence Impact Questionnaire, 24-hour pad weights, daily pad usage and urinalysis at all visits. Urodynamic studies were performed at the 6-week and 3-month visits. RESULTS: Three subjects had uncomplicated urinary tract infections during followup. No other adverse effects occurred. Statistically and clinically significant decreases greater than 50% were seen in virtually all outcome measures at all visits up to 3 months. Most subjects showed signs of recurrent UUI by 6 months. All subjects reported remarkable subjective improvement in incontinence. No significant changes in maximal cystometric capacity, maximal detrusor pressure, peak flow or post-void residual volumes were seen. CONCLUSIONS: Botulinum-A toxin can significantly decrease urge urinary incontinence and improve quality of life for 3 months after injection. Additional studies are needed to determine ideal doses, dosing intervals, safety and cost effectiveness of this therapy. PMID- 15538258 TI - Eliciting preferences for drug treatment of lower urinary tract symptoms associated with benign prostatic hyperplasia. AB - PURPOSE: There are 2 main medical preparations available for lower urinary tract symptoms resulting from benign prostatic hyperplasia (BPH). Choosing between an alpha-blocker and a 5alpha-reductase inhibitor requires trade-offs between their attributes or characteristics. We investigated the relative importance of and trade-offs between the attributes of the 5alpha-reductase inhibitor dutasteride and alpha-blockers in community dwelling men using a validated technique. MATERIALS AND METHODS: A discrete choice experiment was administered to 211 men older than 40 years who were randomly selected from the general United Kingdom population. Attributes investigated in the discrete choice experiment were time to symptom improvement, sexual and nonsexual side effects, the risks of acute urinary retention (AUR) and surgery, cost and prostate size decrease. Using regression analysis the relative importance of these attributes, the trade-offs that men are willing to make between these attributes and the willingness to pay for each attribute were estimated. RESULTS: All attributes were important to respondents. The most important attribute was side effects. The least preferred side effects was impotence, followed by decreased libido and dizziness. Respondents were willing to wait 13, 2 and 8 months longer for symptom improvement in exchange for decreased prostate size, and the risks of AUR and surgery, respectively. Men reporting moderate symptoms were less concerned about sexual side effects, time to symptom improvement and the risk of AUR compared with men reporting mild symptoms. CONCLUSIONS: Given the attribute levels of BPH medical treatment, overall community dwelling men preferred the 5alpha-reductase inhibitor over alpha-blockers. In the interests of shared decision making it is important to consider the importance of eliciting the preferences of patients with BPH. PMID- 15538259 TI - Fate of donor kidney: laparoscopic versus open technique. AB - PURPOSE: Laparoscopic donor nephrectomy (LDN) is an increasingly accepted modality for procuring donor kidneys for transplantation. We analyzed and compared the short and long-term outcomes of living transplant allografts from kidneys procured by laparoscopic or open donor (ODN) technique and managed with a single immunosuppression regimen in each group. MATERIALS AND METHODS: Records of recipients who underwent living (laparoscopic or open) donor nephrectomy were reviewed from August 1999 to July 2001 for LDN and from January 1994 to December 1999 for ODN. Patients included were on a single immunosuppression regimen particular to each group. Sirolimus, mycophenolate mofetil and prednisone were given to the LDN group, and calcineurin inhibitor (FK-506 or cyclosporine), mycophenolate mofetil and prednisone were given to the ODN group. Excluded from study were patients with prior kidney transplant or patients not receiving immunosuppression as previously described. Also excluded from study were patients lost to followup before 1 year. Data were retrieved retrospectively from case notes or from the transplant database and analyzed using SAS software (SAS Institute, Cary, North Carolina). RESULTS: A total of 71 patients from the LDN group and 60 patients from the ODN group qualified for the study. Demographic data are comparable in both groups except for the significantly longer followup in the ODN group. Serum creatinine was 2.2 and 1.8 mg/dl at postoperative day 4, 1.3 and 1.3 mg/dl at day 10, and 1.3 and 1.4 mg/dl at 1 month in the LDN and ODN groups, respectively. Time to achieve nadir serum creatinine was 8.7 versus 6.6 days for LDN and ODN groups, respectively (p = not significant). Delayed graft function was noted in 5 of 71 (7%) in the LDN group and 3 of 60 (5%) in the ODN group (p = 0.5). In the LDN group 13 (18%) patients had a serum creatinine of greater than 1.5 mg/dl at postoperative day 30 compared to 6 (10%) in the ODN group (p = 0.06). Mean serum creatinine at 1 year was lower for LDN recipients (p = not significant). But at last followup this difference became statistically significant in favor of LDN. Mean followup was 939 versus 2,046 days for LDN versus ODN, respectively (p <0.0001). Recipient mean hospital stay was 5.2 versus 6.7 days for LDN versus ODN, respectively (p = 0.08). There were 8 of 78 (10.2%) episodes of acute rejection in the LDN group compared to 22% in the ODN group (p = 0.08). The complication rate (ureteral vascular, lymphocele, acute rejection and wound) was 11% in LDN compared to 15% in the ODN group. Long-term graft function, graft survival and patient survival in the LDN group were comparable to the ODN group. CONCLUSIONS: Early graft recovery is slower in LDN allografts, although not statistically significant, but long-term function in the LDN group is significantly better compared to the ODN group. Laparoscopic donor kidneys take longer to achieve nadir serum creatinine, but this does not influence long term outcome and results. PMID- 15538260 TI - Graft function after kidney transplantation from non-heartbeating donors according to maastricht category. AB - PURPOSE: Donor shortages have led to some groups using alternative sources such as non-heartbeating donors (NHBDs). Kidneys from NHBDs suffer from warm ischemia at cardiac arrest which is reflected by acute tubular necrosis of the allograft, resulting in a period of delayed graft function. NHBDs are categorized by the circumstances surrounding the agonal events of death which reflect differences in the likelihood of ischemic injury to the kidney. In this study we determined the impact of ischemic injury on the renal function of kidneys procured from different categories of NHBDs. MATERIALS AND METHODS: From 1998 to 2003, 144 kidneys were procured from 72 NHBDs resulting in 93 transplants characterized into Maastricht categories II, III and IV NHBD renal transplants. Renal function after transplant was evaluated from the last dialysis until discharge from hospital, and then at 3 monthly intervals thereafter. RESULTS: Primary warm ischemic time is more prolonged in the uncontrolled donor (category II) than controlled donor (category III greater than IV). Delayed graft function occurs more frequently (Maastricht category II 83.8%, III 67.4% and IV 0%, ANOVA p <0.05) and the return to normal function is more prolonged in uncontrolled donors. This is illustrated by the greater incidence of acute tubular necrosis (Maastricht category II 81.1%, III 65.2% and IV 50.0%, ANOVA p = nonsignificant) in the kidney allograft. There was no difference in year 1 allograft survival (Maastricht category II 83.9%, III 92.5% and IV 100%, ANOVA p = nonsignificant). CONCLUSIONS: Early graft function is poorest in kidneys derived from Maastricht category II donors and best in category IV with III in-between. However, after 3 months the function of kidneys from all donors is the same. PMID- 15538261 TI - Effect of the use of internal iliac artery for renal transplantation on penile vascularity and erectile function: a prospective study. AB - PURPOSE: We determined the effects of the use of internal iliac artery for renal transplantation on erectile function and penile hemodynamics in prospective fashion. MATERIALS AND METHODS: The study comprised 50 patients with chronic renal failure on hemodialysis without associated vascular risk factors. All patients were evaluated before and after renal transplantation with 6-month followup. All grafts were revascularized with end-to-end anastomosis to the right internal iliac artery. Before and after transplantation the patients were evaluated by detailed history, including the International Index of Erectile Function, clinical examination, laboratory investigations and pharmacodynamic penile color duplex ultrasonography. RESULTS: Erectile dysfunction was reported by 28% of the patients with chronic renal failure. This incidence becomes 26% after transplantation. Based on questionnaires post-transplantation erectile function compared to pre-transplant status was improved, deteriorated or remained static in 40%, 18% and 42% of transplant recipients, respectively. The International Index of Erectile Function showed no differences between pre transplantation and posttransplantation sexual function except in the domain of sexual desire. In comparison to preoperative penile indices, there was a significant decrease in penile arterial blood flow in the 2 cavernous arteries (p <0.05). However, none of the patients had penile arterial insufficiency. CONCLUSIONS: Renal transplantation has varying effects on erectile function. In the majority of cases it has no negative effect on the quality of erection. In the absence of associated vascular risk factors unilateral interruption of the internal iliac artery decreases arterial penile blood flow but not to a degree that compromise erectile function. PMID- 15538262 TI - Surgical management of large adrenal masses with or without thrombus extending into the inferior vena cava. AB - PURPOSE: Surgical extirpation is the only curative treatment for large adrenal masses with or without thrombus extending into the inferior vena cava. However, occasionally complex surgical techniques are required, including venovenous bypass or cardiopulmonary bypass (CPB). Additionally, applying techniques used for organ transplantation can provide better exposure with less blood loss to allow milking of the thrombus downward, limiting the need for bypass. MATERIALS AND METHODS: Ten patients underwent surgery for large adrenal masses using these techniques. Five patients had thrombi extending into the inferior vena cava, causing Budd-Chiari syndrome in 1. A classification system was proposed for adrenal masses associated with venous thrombus. RESULTS: Median patient age was 51 years. Surgery was completed successfully in all patients. Only 1 patient with an adherent intra-atrial thrombus required CPB. Mean blood loss was 450 ml (range 50 to 1,500) except in the patient who required CPB. Postoperative complications occurred in 2 patients. One patient died on the postoperative day 7 of a presumed pulmonary emboli. Pneumothorax and empyema following traumatic line placement developed in the other patient. Nine patients (90%) were free of disease at a median followup of 18 months (range 10 to 84). CONCLUSIONS: Applying transplant techniques in the surgical extirpation of large adrenal masses with or without tumor thrombus affords curative surgery enhanced access and vascular control, and decreases the requirement for venovenous bypass and/or CPB with less morbidity. It also provides acceptable midterm survival and quality of life. PMID- 15538263 TI - Kidney gene database: a curated and integrated database of genes involved in kidney disease. AB - PURPOSE: We have created a curated and integrated database, the Kidney Gene Database (KGDB) (http://www.urogene.org/kgdb) that contains current information about genes or genomic loci involved in human kidney disease. MATERIALS AND METHODS: Genes that undergo molecular, genetic or epigenetic events that affect kidney function are identified through a biomedical literature search and catalogued in the database. RESULTS: Events that are currently screened for are gene amplification, mutation, deletion, polymorphism, loss of heterozygosity, DNA methylation and DNA hypomethylation. In addition, genes that are uniquely expressed in the kidney, as determined by analyzing the expressed sequence tags database and the Serial Analysis of Gene Expression database, are also included in KGDB. For each gene KGDB provides basic information about the gene product, a tissue type gene expression profile, links to protein, mRNA and genomic DNA sequence information, relevant literature citations and cross-references to other databases. CONCLUSIONS: We present KGDB, which is to our knowledge the first curated and integrated database of genes involved in human kidney disease. KGDB is free, widely accessible and easy to use, and it provides a wealth of relevant information. In addition, KGDB will be continuously updated every 6 months to include new information published in the biomedical literature or in gene expression databases. We envision that KGDB will serve as a valuable resource for scientists and clinicians. PMID- 15538264 TI - Prospective, randomized, crossover comparison of sublingual apomorphine (3 mg) with oral sildenafil (50 mg) for male erectile dysfunction. AB - PURPOSE: We established the efficacy and safety of sublingual apomorphine compared with oral sildenafil in comparable groups of patients with erectile dysfunction (ED). MATERIALS AND METHODS: This prospective, randomized, crossover study included 77 heterosexual men with ED of various etiologies and severities. A total of 62 men were randomized but only 34 were evaluable for efficacy and tolerability. The study started with a run-in period of 2 to 4 weeks. The first 4 weeks of treatment were followed by a washout period of 4 weeks, after which patients changed to the alternate treatment for an additional 4-week period. The sequence of the 2 treatments was established by a randomization list in blocks in closed packets. The primary efficacy end point was the percent of attempts resulting in erection firm enough for intercourse. Additional variables were the percent of attempts resulting in intercourse and improvement in ED, as evaluated by the erectile function domain score of the International Index of Erectile Function questionnaire. RESULTS: Sildenafil was significantly more effective than apomorphine in regard to the percent of attempts resulting in erection firm enough for intercourse (85% vs 44%, p <0.0001) and actually resulting in intercourse (81% vs 43%, p <0.0001) as well as erectile function evaluated by the erectile function domain score of the International Index of Erectile Function (p <0.001). The incidence of adverse events was not significantly different for the 2 drugs. Although the number of patients was small, this study had strong statistical power due to the striking difference in results. CONCLUSIONS: Sildenafil was significantly more effective than apomorphine for ED. No statistical difference in adverse events was noted. PMID- 15538265 TI - The role of sildenafil in the treatment of erectile dysfunction in patients with pelvic fracture urethral disruption. AB - PURPOSE: Erectile dysfunction (ED) is a common sequel of pelvic fracture urethral disruption (PFUD). After repair of the urethral injury ED may be the most devastating long-term effect for the patient. Some patients with ED may regain normal erectile function. We prospectively studied the response to sildenafil and the erectile function of patients with ED due to PFUD. MATERIALS AND METHODS: The erectile function of patients referred to us with PFUD for urethroplasty were prospectively evaluated before surgery. Patients underwent nocturnal penile tumescence testing and, if results were abnormal, penile duplex ultrasonography with intracavernous injection and arteriography were performed to diagnose the etiology of ED. Patients were questioned about erectile function every 3 months after surgery and if they complained of ED they were offered 100 mg sildenafil. Patients were followed for at least 18 months after surgery. RESULTS: A total of 29 consecutive patients were evaluated and 22 (76%) of them had ED before surgery. Sufficient followup was available for 15 of the patients. Overall 47% of these patients responded favorably to sildenafil. Of the patients 60% with neurogenic ED and 20% of those with arterial ED responded to this treatment. In 33% of the patients ED resolved within the followup period. All patients with spontaneous resolution of ED previously responded to sildenafil (71% of sildenafil responders). CONCLUSIONS: In patients with ED due to PFUD, those with neurogenic ED are more likely to respond to sildenafil than those with arterial damage. Favorable response to sildenafil may predict spontaneous resumption of normal erectile function over time. PMID- 15538266 TI - Cystectomy and orthotopic ileal neobladder: the impact on female sexuality. AB - PURPOSE: January 1986 and September 2003 cystectomy and formation of an ileal neobladder were performed in 86 female patients. In this retrospective study we focused on the impact of orthotopic reconstruction on female sexuality. MATERIALS AND METHODS: To assess female sexuality the standardized self-reporting instrument female sexual function index was mailed to 44 patients who were 70 years or younger at cystectomy with a recurrence-free followup of greater than 1 year, no pelvic irradiation and no concomitant diseases impairing sexual functions. The questionnaire analyzes 6 domains (desire, arousal, lubrication, orgasm, satisfaction and pain) with 19 items. It was returned by 29 patients (65.9%) with a median age of 65.0 years. The indication for cystectomy was benign disease in 8 cases and malignancy in 21. RESULTS: Factors influencing female sexuality were age younger than 60 years, benign disease, partnership at surgery and current partnership. Clean intermittent catheterization, urinary stress incontinence and hormonal therapy did not affect the results. The 11 of 17 patients who remained sexually active after cystectomy even had slight improvement in all female sexual function items. Six patients ceased to be sexually active postoperatively due to erectile dysfunction or partner death. One patient with interstitial cystitis became sexually active following cystectomy due to the loss of pelvic pain. Another 12 patients remained sexually inactive postoperatively. CONCLUSIONS: All aspects of female sexuality may remain unchanged following cystectomy and ileal neobladder formation as long as sexual activity is not ceased due to other reasons. Even fertility can be preserved when the internal genitalia do not have to be removed. PMID- 15538267 TI - Spermatic cord anesthesia block for scrotal procedures in outpatient clinic setting. AB - PURPOSE: We evaluated the safety and effectiveness of spermatic cord anesthetic block (SCAB) as the only method of anesthesia for bilateral simple orchiectomy in an outpatient clinic setting. MATERIALS AND METHODS: The study included 141 consecutive bilateral simple orchiectomy (BSO) procedures performed at Atlanta Veterans Affairs Medical Center during a 33-month period. All procedures were performed in an outpatient clinic setting using SCAB as the only method of anesthesia. The anesthetic solution consisted of an equal mixture of 1% lidocaine with epinephrine at 1:100,000 and 0.25% bupivacaine. A 10-point visual analog pain scale was used to assess pain/discomfort at baseline, during SCAB instillation and during BSO. Postoperatively overall patient impression/satisfaction with SCAB as a method of anesthesia was determined. RESULTS: Nine of the 141 available cases (6.4%) were excluded from study. The remaining 132 cases were included in data analysis. Mean patient age was 75.4 years (range 44 to 86). A total of 76 patients (57.6%) were receiving luteinizing hormone releasing hormone agonist therapy at the time of the procedure. The mean time needed to perform SCAB and BSO was 3.9 (range 2 to 6) and 33.0 minutes (range 12 to 70), respectively. The average volume of anesthetic solution was 20.1 ml per case (range 10 to 32). The mean pain score was 0.36 (range 0 to 8), 1.96 (range 0 to 8) and 0.33 (range 0 to 5) at baseline, and during SCAB and BSO, respectively. Of the patients 102 (77.3%) underwent a painless procedure (pain score = 0), 29 (21.9%) experienced transient pain that was mild in nature (pain score 4 or less) and only 1 (0.7%) had a pain score of 5. Pain scores were similar in patients who were and were not receiving luteinizing hormone releasing hormone agonists at baseline (p = 0.36), during SCAB instillation (p = 0.89) and during BSO (p = 0.36). The overall impression/satisfaction with SCAB as a method of anesthesia was rated as highly satisfactory by 91.7% of patients and satisfactory by 8.3%. There were no intraoperative adverse events related to SCAB. CONCLUSIONS: SCAB is a simple, safe and highly effective method of anesthesia for scrotal procedures. It allows such procedures to be performed in an outpatient clinic setting, offering convenience and the potential for cost savings. PMID- 15538268 TI - Geographic variation across veterans affairs medical centers in the treatment of early stage prostate cancer. AB - PURPOSE: We investigated geographic variation in the treatment of early stage prostate cancer in a national sample of veterans after widespread adoption of the prostate specific antigen test. MATERIALS AND METHODS: Our sample consisted of 16,352 cases from the Veterans Affairs Central Cancer Registry that were diagnosed between January 1997 and December 1999 with stage I or II prostate cancer. We used a 2-stage nested logit model to compare surgery, radiation therapy and noncurative treatment among 4 geographic regions of the United States. RESULTS: Multivariate analysis showed that patients in the West (referent group) had a higher OR of undergoing surgery than radiation compared with the Northeast, South or Midwest (OR 0.77, 95% CI 0.67 to 0.87, OR 0.86, 95% CI 0.76 to 0.98 and OR 0.75, 95% CI 0.64 to 0.87, respectively. Black men, men with lower grade and higher stage tumors, and unmarried men were less likely to undergo curative treatment and less likely to undergo surgery than radiation. CONCLUSIONS: Geographic variation persists in patterns of care in men with early stage prostate cancer. However, this variation is limited to the choice between surgery and radiation rather than to the choice between curative and noncurative treatment. PMID- 15538269 TI - Validity of primary care patient self-report of abnormal (but not cancer) prostate biopsy results. AB - PURPOSE: We assessed the validity of patient self-report of abnormal (but not cancer) prostate biopsy results, ie, atypical cells or prostatic intraepithelial neoplasia. MATERIALS AND METHODS: We compared survey responses and pathology records for 156 men from a multicenter, prospective cohort study, which was designed to evaluate the impact of a suspicious prostate cancer screening test followed by benign prostate biopsy. RESULTS: Of the 156 subjects 45 (29%) reported having had atypical cells or prostatic intraepithelial neoplasia and 38 (24%) had a pathology report with atypia or prostate intraepithelial neoplasia. The sensitivity of the self-report was 87%, specificity was 90%, positive predictive value was 73% and negative predictive value was 95%. The kappa statistic was 0.7. CONCLUSIONS: Men in this study reported this complex result accurately. However, it was concerning that 5 men (13%) appeared unaware of an abnormal prostate biopsy result, especially given the substantial residual risk of prostate cancer conferred by this result. PMID- 15538270 TI - Prepubertal testis tumors: actual prevalence rate of histological types. AB - PURPOSE: Tumor registries, urological textbooks and literature surveys all assert that yolk sac tumors are the most common primary testicular tumors in boys 12 years and younger. In contrast, several individual institutions have reported that benign tumors are more common than malignant tumors. To clarify these discordant findings, we surveyed the primary pathology records from 4 major pediatric centers. MATERIALS AND METHODS: The pathology records of the contributing centers were culled for primary testicular masses in boys 12 years and younger. Older boys and those with either paratesticular tumors or leukemia were excluded. The prevalence of each histological subtype was calculated from the pooled cases. RESULTS: A total of 98 patients met our criteria. Only 15% had yolk sac tumors. Teratomas comprised 48% of the tumors (mature 44%, immature 4%). Epidermoid cysts were found in another 14% of patients. Gonadal stromal cell tumors represented 13% of the total, divided among granulosa cell (5%), Leydig cell (4%), Sertoli cell (3%) and mixed gonadal stromal cell (1%). Other pathology, including cystic dysplasia (2), lymphoma (4), inflammatory pseudotumor (1) and gonadoblastoma (2), made up 9% of the total number of cases. CONCLUSIONS: We found that benign lesions represent the majority of primary testis tumors (74%), with the most common histological type being teratoma (48%). The reported high prevalence rates of prepubertal yolk sac tumors probably results from a reporting bias, since benign tumors are less likely to be submitted to tumor registries. Therefore, the primary operative approach to the majority of testis tumors in boys 12 years and younger should entail testis sparing surgery. Orchiectomy should be reserved for histologically confirmed malignancy based on increased preoperative alpha-fetoprotein and/or frozen section analysis of the tumor. PMID- 15538271 TI - Spermatic cord torsion with preserved testis perfusion: initial anatomical observations. AB - PURPOSE: Despite the high sensitivity and specificity of scrotal ultrasonography, there continue to be reports of missed torsion. These "false-negative" scans are attributed to technical factors and intermittent torsions. We hypothesize that patients with specific anatomical configurations maintain testicular blood flow for prolonged periods, and, therefore, will have flow on ultrasound despite concurrent torsion. MATERIALS AND METHODS: Patient charts and scrotal ultrasounds were reviewed to identify patients younger than 18 years who underwent scrotal exploration between January 1998 and January 2003 for acute scrotum. Patients who underwent radiological evaluation before scrotal exploration were the main focus of this study. Operative reports were reviewed for specific anatomical details. RESULTS: A total of 61 patients underwent scrotal exploration for acute scrotum during the study period. Of these patients 14 had torsion confirmed intraoperatively and a preoperative scrotal ultrasound available. Four of these 14 patients had normal testicular blood flow on ultrasound but had testicular torsion confirmed intraoperatively. Operative findings suggest that these patients have specific anatomical characteristics. CONCLUSIONS: Testis perfusion can be maintained for a prolonged period in the presence of testicular torsion. Anatomical variability may account for differences in the duration of viability of the torsed testis. A high index of suspicion must be maintained to avoid missing the diagnosis of testicular torsion in these challenging cases. PMID- 15538272 TI - Surgical reconstruction of cloacal malformation can alter bladder function: a comparative study with anorectal anomalies. AB - PURPOSE: Patients with cloacal anomalies often suffer bladder dysfunction with recurrent urinary infections and incontinence. We examined the effect of surgical reconstruction by posterior sagittal approach and total urogenital mobilization in either causing or worsening bladder dysfunction. MATERIALS AND METHODS: Between August 2000 and December 2002 all new patients with cloacal anomalies were prospectively studied to assess the effect of surgical reconstruction by posterior sagittal approach and total urogenital mobilization on bladder function. A comparable group of patients with anorectal malformation (ARM) were studied as comparative controls to assess the effect of posterior sagittal approach without urogenital surgery. Structural anatomy was defined by radiology. Natural filling urodynamics via suprapubic catheter were performed in all infants at 0.2 to 9 months (mean 3) before surgical reconstruction. This assessment was repeated 6 to 24 months (mean 14.8) after surgery, and changes in bladder function were determined by comparative statistics. RESULTS: A total of 10 patients with cloacal anomalies (5 with short [less than 3 cm] and 5 with long common channel [greater than 3 cm]) and 20 patients with anorectal malformation were consecutively studied. At presentation bladder dysfunction was present in 9 of 10 patients with cloacal anomalies and in 12 of 20 patients with ARM. After surgery there was significant deterioration in bladder function in half of the cloacal group (5 of 10 patients, p = 0.04) and in 1 of 20 patients with ARM (p = 0.7). Of the 5 patients with cloacal anomalies who had deterioration of bladder function urodynamic pattern of detrusor overactivity changed to inadequate (atonic) bladder in 4, all of whom had a long common channel at presentation. CONCLUSIONS: Patients with cloacal malformation have a high incidence of innate bladder dysfunction. However, surgical reconstruction by total urogenital mobilization can cause further deterioration of bladder function, particularly in the group with a long common channel. Urodynamic assessment is necessary to detect bladder dysfunction in these patients. PMID- 15538273 TI - Results of 265 consecutive proximal hypospadias repairs using the Thiersch-Duplay principle. AB - PURPOSE: We review the evolution of the concept of tubularization of the urethral plate and our results in the repair of proximal hypospadias. MATERIALS AND METHODS: A total of 281 children born with proximal hypospadias underwent Thiersch-Duplay urethroplasty with or without a midline incision of the urethral plate between 1989 and 1998. Followup data were available in 265 children. RESULTS: Excellent functional and cosmetic results were achieved in 88.7% of the patients. The use of either a dartos or tunica vaginalis flap to waterproof the urethral suture line resulted in a decrease in the fistula rate from 17% to 1.8%. CONCLUSIONS: The principles of Thiersch-Duplay urethroplasty represent the basic foundation for surgical techniques that use the urethral plate to construct a urethral tube. The use of this principle in the repair of proximal hypospadias compares favorably with other methods. PMID- 15538274 TI - Urinary continence is well preserved after total urogenital mobilization. AB - PURPOSE: Total urogenital sinus mobilization (TUM) has been advocated as a successful technique in the repair of common urogenital sinus anomalies. To our knowledge the long-term effects on continence, voiding pattern and bowel control have not been published. We present our experience and assessment regarding these issues. MATERIALS AND METHODS: We retrospectively reviewed our experience with TUM. Medical records were reviewed for each patient along with long-term followup by telephone questionnaire and clinical visits. Uroflowmetry was performed in recent patients who were old enough to cooperate in the study. RESULTS: A total of 13 females underwent TUM. Medical records were reviewed for all patients. Patients were divided into 2 groups. Group 1 consisted of 7 patients who underwent surgery before the age of achieving continence, and group 2 consisted of 6 patients who were incontinent before surgery. One patient in group 2 was lost to long-term followup. Two children in group 1 have enuresis, a 27-month-old with day and nighttime enuresis and a 3-year-old with only nocturnal enuresis. All patients in group 2 were continent immediately postoperatively. CONCLUSIONS: TUM preserves urinary continence immediately postoperatively among patients who are continent before surgery. Furthermore, it does not appear to delay the natural development of continence among patients undergoing surgery before the age of toilet training. PMID- 15538275 TI - Cyclooxygenase-2 inhibitor preserves medullary aquaporin-2 expression and prevents polyuria after ureteral obstruction. AB - PURPOSE: Renal obstruction causes impairment of urinary concentrating ability, partly by decreasing aquaporin-2 (AQP-2) water channel level in the collecting ducts. We reported previously that ureteral obstruction induced cyclooxygenase-2 (COX-2) in the medullary collecting duct cells by increased mechanical stretch. In this study we investigated whether AQP-2 decrease after obstruction was regulated by COX-2. MATERIALS AND METHODS: Sprague-Dawley rats were subjected to bilateral ureteral obstruction for 24 to 48 hours. During obstruction rats were given NS398, a COX-2 specific inhibitor, by oral gavage (2 mg/kg per day). COX-2 and AQP-2 levels were assessed in the inner medulla using Western blot. Urine output was measured after releasing obstruction to assess the degree of polyuria. RESULTS: With obstruction COX-2 protein levels increased and AQP-2 levels decreased in the inner medulla. Corresponding to the loss of AQP-2, urine output increased 4.2-fold after obstruction. The obstructed rats receiving NS398 exhibited significant preservation of AQP-2 level (72% of control), as well as significant normalization of urine output. The sham operated rats receiving NS398 exhibited an increased amount of AQP-2 protein level. CONCLUSIONS: These findings suggest that COX-2 mediated prostaglandin has an important role in the down regulation of AQP-2 water channel level in the medullary collecting duct cells after ureteral obstruction. PMID- 15538276 TI - Laparoscopic transuterine fetal vesicostomy: a feasibility study. AB - PURPOSE: We evaluate the feasibility of applying minimally invasive techniques for fetal vesicostomy. We also evaluate whether transuterine fetal vesicostomy can be performed laparoscopically. MATERIALS AND METHODS: A total of 25 pregnant ewes were time dated at approximately 90 days of gestation. With the animals under general anesthesia a low open abdominal incision was made and the uterus was brought out through the incision. With a 14 gauge needle the amniotic sac was filled with 1 to 2 l warm glycine. Three to 4, 5/12 blunt tip balloon trocars were placed in the uterus. Using laparoscopic techniques, a low transverse incision was made in the fetal abdomen, the bladder was opened at the dome and 2 running sutures were placed approximating the fetal abdominal wall to the edge of the fetal bladder. The trocar sites in the uterus were closed, and the maternal abdominal incision was closed. RESULTS: Of the 25 pregnant ewes the technique was developed in the initial 15. In the subsequent 10 animals the complete procedure was accomplished successfully. Following these 10 procedures 5 abortions occurred on postoperative day 2, and there was 1 intrauterine fetal demise. Three fetuses were alive and delivered by cesarean section on postoperative days 10, 30 and 31. In the first fetus in which we used an interrupted suture for the vesicostomy a large hernia was noted at the vesicostomy site. The other 2 fetuses had a patent, well healed vesicostomy and were alive at cesarean section delivery on postoperative days 10 and 31. The last fetus was allowed to deliver at term by standard vaginal delivery. The fetus was alive and well, and the vesicostomy had strictured down to a pinhole in size, which was not unexpected as it was not an obstructed model. CONCLUSIONS: Although technically challenging, transuterine laparoscopic fetal vesicostomy is technically feasible in the ewe model. Continued evaluation of this technique should include intensive fetal monitoring and the use of tocolytics to decrease the incidence of spontaneous abortion. PMID- 15538277 TI - A 7-year experience of genetic males with severe phallic inadequacy assigned female. AB - PURPOSE: We examine the hypotheses underlying the clinical decision to assign female gender at birth in male neonates with severe phallic inadequacy. MATERIALS AND METHODS: A total of 18 genetic males with severe phallic inadequacy were assessed longitudinally for physical, social, psychological and sexual identity development. Diagnoses included cloacal (11) and classic (1) exstrophy, partial androgen resistance (3), mixed gonadal dysgenesis (2) and penile agenesis (1). Neonatal sex assignment was female in 15 and male in 3 patients. RESULTS: All patients demonstrated marked male typical behaviors and interests. Of the 15 female assigned patients 1 died, 1 refused to declare sexual identity or orientation, 1 converted to male before initial evaluation, 1 was reassigned male by the parents and 5 others declared male sexual identity. Thus, of 17 living patients 10 live as males and 6 as females. Of patients 17 years or older only those living as male lived independently. The 4 oldest patients living as male but only 1 patient living as female would discuss sexual orientation-all 5 declared orientation toward females, and 3 of these 4 males had girlfriends or were married. The sexually undeclared patient lived in a residential treatment center because of major behavioral difficulties since age 15. Only patients living as male had dated. CONCLUSIONS: This longitudinal study implies that males with severe phallic inadequacy reared male and those reared female but converting to male can have functional psychosocial developmental trajectories. Those reared female have a realistic likelihood of recognizing male sexual identity and converting to male. Those not converting to male appear to have less successful psychosocial developmental trajectories. PMID- 15538278 TI - Successful outpatient management of the nonpalpable intra-abdominal testis with staged Fowler-Stephens orchiopexy. AB - PURPOSE: Previous reports of orchiopexies have shown an 85.9% success rate for staged Fowler-Stephens orchiopexy (FSO). We review our experience with nonpalpable testes (NPTs). MATERIALS AND METHODS: A total of 119 patients with 128 NPTs were treated at our institution between 1994 and 2001. Atrophic testes or "nubbins" were removed. Once an intra-abdominal testis was identified the peritoneum was opened and firm traction was placed on the gonad. If feasible, primary orchiopexy was completed. Otherwise, staged Fowler-Stephens orchiopexy was performed. RESULTS: Primary orchiopexy was performed in 28 testes in 27 patients without division of the spermatic vessels. An atrophic nubbin was removed in 45 patients. All children with bilateral NPTs had at least 1 viable intra-abdominal testis found at surgery. Staged FSO was performed in 55 testes in 47 children. The second stage was performed at a median of 3.5 months after initial ligation of the spermatic vessels. Median followup was 1 year and mean followup was 9 months. Five patients were lost to followup. Successful surgery was defined as a dependent scrotal location and testis size equivalent to the contralateral mate. The overall success rate for the primary orchiopexy group was 100%. In the staged FSO group 1 patient had an atrophic testis at 1-year followup, yielding an overall success rate of 98%. CONCLUSIONS: A high degree of success can be obtained for children with intra-abdominal testes. Mobility of the testis on exploration is a good indicator that the testis can be managed with primary orchiopexy without division of the vessels. If primary orchiopexy cannot be performed, excellent results are achieved with a staged FSO. PMID- 15538279 TI - The influence of orchiopexy on serum inhibin B level: relationship with histology. AB - PURPOSE: Since inhibin B is the endocrine marker of spermatogenesis, basal inhibin B levels may reflect germ cell status in children. The aims of this study were to determine the changes in endocrine parameters after orchiopexy in patients with cryptorchidism and to compare these findings with testicular biopsy parameters. MATERIALS AND METHODS: A total of 27 boys with undescended testis were included in this study. Inguinal orchiopexy was performed in all patients and 15 underwent testicular biopsy at orchiopexy. Spermatogonia per tubular transverse section and fertility index values were determined. Before and 6 months after orchiopexy serum basal inhibin B and other serum hormone levels were measured in all patients. RESULTS: Mean serum basal inhibin B levels significantly increased 6 months after successful orchiopexy (p = 0.001). However, inhibin B level did not increase in patients who had a low testicular biopsy score. Other reproductive hormone levels did not change after orchiopexy. CONCLUSIONS: Basal inhibin B level could be used as a followup parameter after orchiopexy. If basal inhibin B level does not increase in the postoperative period, the amount of germ cells in the testis may be too low or the orchiopexy might not have been implemented appropriately. PMID- 15538280 TI - Urine cytology in the evaluation of upper tract urothelial lesions. PMID- 15538281 TI - Computerized tomography criteria of renal cell carcinoma. PMID- 15538282 TI - Association of the cyclin D1 gene G870A polymorphism with susceptibility to sporadic renal cell carcinoma. AB - PURPOSE: Cyclin D1 (CCND1) mRNA is alternatively spliced to produce 2 transcripts (transcript-a and transcript-b), which may be modulated by a G870A single nucleotide polymorphism at the conserved splice donor site of exon 4. Previous studies have suggested a significant association between the CCND1 genotype and the development of various cancers. We explored the possible association between this polymorphism and the onset or disease statue of sporadic renal cell carcinoma (RCC). MATERIALS AND METHODS: The CCND1 G870A genotype was determined in 191 RCC cases and in 400 controls by polymerase chain reaction restriction length polymorphism analysis. RESULTS: Subjects with the AA genotype were at 1.70 fold significant higher risk for RCC than those with the GG genotype (age and sex adjusted OR 1.70, 95% 95% CI 1.03 to 2.82, p = 0.039). In addition, the A allele had a gene dose effect in increasing the risk of RCC (adjusted OR 1.30, 95% CI 1.01 to 1.67, p = 0.045). For tumor stage no significant difference in genotype frequency was found (p = 0.646). CONCLUSIONS: These data suggest that the CCND1 variant A allele may be a genetic susceptibility factor with a recessive or gene dose effect for the onset of sporadic RCC. More extensive and larger studies are required to clarify whether the CCND1 genotype is more specifically involved in the onset of a histological subset of RCC or RCC at a younger age. PMID- 15538283 TI - Use of prostate specific antigen to measure bladder tumor growth in a mouse orthotopic model. AB - PURPOSE: Animal orthotopic tumor models are commonly used in bladder cancer studies. However, to our knowledge there is currently no accurate method to quantify tumor growth inside the bladder in living animals. We used prostate specific antigen (PSA) as a marker to cope with this limitation. MATERIALS AND METHODS: Infectious but replication incompetent retroviral particles carrying PSA coding sequence were constructed and infected into MB49 cells, a mouse bladder transitional cell carcinoma line of C57BL/6 origin. Syngeneic mice were intravesically implanted with the novel MB49-PSA transfectants. Tumor burden was evaluated by enzyme-linked immunosorbent assay measurement for PSA in urine and bladder tissues. RESULTS: The MB49-PSA line actively secreted PSA in culture as well as in urine (18 to 2,062 pg/ml) depending on tumor mass. Immunofluorescence staining confirmed PSA expression in MB49-PSA derived orthotopic tumors. Urinary PSA production paralleled tumor growth and was detectable prior to the development of a palpable tumor. Although urinary PSA did not tightly correlate with tumor mass, all bladders (total of 16 tested) weighing 34 mg or greater (18 to 21 mg for age and sex matched normal bladders) showed 18 pg/ml or greater urinary PSA. In contrast, bladder tissue PSA correlated more with tumor mass in general and it was measurable even before the detection of urinary PSA. This MB49 PSA orthotopic tumor model also demonstrated its usefulness for evaluating the antibladder cancer agents gemcitabine and mitomycin. CONCLUSIONS: This novel MB49 PSA line may serve as a useful tool for bladder cancer study because its growth inside the bladder can be noninvasively measured in living animals even during early stages of tumor growth. PMID- 15538284 TI - Transforming growth factor-beta1 induced myofibroblasts regulate LNCaP cell death. AB - PURPOSE: Reactive stroma represents a generic wound response phenomenon, which has been identified in areas of tissue injury and carcinogenesis. To determine whether reactive stroma influences prostate tumor cell growth 3 primary prostate stromal cell lines were treated with transforming growth factor-beta1 (TGF-beta1) to induce the reactive stroma phenotype and then co-cultured with LNCaP cells. MATERIALS AND METHODS: Flow cytometry was performed in LNCaP cells that had been co-cultured with induced reactive stroma or control stroma and an index of cell death and proliferation was obtained. Using the previously described 3 way differential reactive stroma xenograft tumor model consisting of LNCaP cells, stromal cells and Matrigel (Collaborative Research, Bedford, Massachusetts) LNCaP cell apoptosis was evaluated using TUNEL staining in a background of varying degrees of reactive stroma. RESULTS: Flow cytometric analysis revealed that LNCaP cells co-cultured with TGF-beta1 induced stromal cells demonstrated a significantly decreased rate of cell death compared with controls (p <0.001). In an animal model LNCaP cells of the 3 way xenograft constructs treated with TGF beta1 latency associated peptide, an inhibitor of TGF-beta1, showed increased apoptosis by TUNEL staining (p <0.001). Double label immunohistochemistry analysis demonstrated that TGF-beta1 induced stromal cells had an increased proportion of myofibroblasts, the identifying cell type of reactive stroma. Furthermore, the degree of reactive stroma inversely corresponded to the degree of LNCaP cell death. CONCLUSIONS: These findings indicate that reactive stroma influences prostate cancer cell growth and warrant investigation of the regulatory mechanisms between reactive stroma and prostate cancer cells. PMID- 15538285 TI - Phytoestrogens derived from Belamcanda chinensis have an antiproliferative effect on prostate cancer cells in vitro. AB - PURPOSE: Phytoestrogens are nonsteroidal plant derived compounds with estrogenic activity that have been implicated in protecting against prostate cancer progression. We hypothesized that these compounds would alter cell number and increase the ability of antiandrogens to induce cell death in prostate cancer cells. MATERIALS AND METHODS: RWPE-1, LNCaP and PC-3 cells were treated with or without an extract of Belamcanda chinensis, 2 purified phytoestrogens derived from this extract (irigenin and tectorigenin) and the antiandrogen bicalutamide. We assessed the effect on cell number, proliferation and apoptosis. RESULTS: Phytoestrogens (50 to 100 microM) and bicalutamide (10 to 50 microM) alone decreased the cell number in all 3 cell lines. Phytoestrogens (50 microM) combined with bicalutamide (10 microM) further decreased the number of RWPE-1 and PC-3 cells compared to these agents alone. Tectorigenin and irigenin inhibited the proliferation of RWPE-1, LNCaP and PC-3 cells, causing G1 arrest and the induction of p21WAF1 or p27 protein expression, whereas bicalutamide induced apoptosis in a dose dependent manner in all 3 cell lines. Phytoestrogens did not have antiandrogenic activity. CONCLUSIONS: These in vitro studies demonstrate a role for tectorigenin and irigenin in regulating prostate cancer cell number by inhibiting proliferation through cell cycle regulation. PMID- 15538286 TI - Cyclophosphamide induced cystitis alters neurotrophin and receptor tyrosine kinase expression in pelvic ganglia and bladder. AB - PURPOSE: We examined neurotrophin and receptor tyrosine kinase (Trk) expression in the bladder and major pelvic ganglia (MPG) after cyclophosphamide induced cystitis in rats. MATERIALS AND METHODS: The bladder and MPG were used in immunohistochemical studies, enzyme-linked immunoassays and Western blots for nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), TrkA and TrkB. Bladder postganglionic MPG cells were labeled by tracing techniques. RESULTS: NGF and BDNF expression was decreased in the bladder of all rats after cystitis (p < or =0.001). NGF and BDNF expression was increased in the MPG in male rats with cystitis (p < or =0.01). Cells expressing TrkA and TrkB immunoreactivity (IR) increased 78% to 81% in the MPG in male rats with cystitis. TrkA-IR or TrkB-IR bladder postganglionic cells increased 50% to 74% with cystitis. Cystitis increased TrkA-IR 5 to 10-fold and TrkB-IR 10 to 12-fold in detrusor muscle. TrkA-IR and TrkB-IR were prominent in control urothelium but decreased with cystitis. After cystitis TrkB-IR nerve fibers and TrkA-IR cellular infiltrates were more apparent compared to controls. CONCLUSIONS: Cystitis decreases bladder NGF and BDNF expression, whereas MPG expression is increased. This change may reflect neurotrophin release at the bladder and retrograde transport to the MPG. TrkA-IR and TrkB-IR are increased in bladder postganglionic cells and bladders with cystitis. This increase may reflect a shift in Trk staining from urothelium to detrusor muscle and nerve fibers with cystitis. Neurotrophin/Trk interactions in the bladder and MPG may contribute to bladder overactivity with cystitis. PMID- 15538287 TI - Inflammation of the rat prostate evokes release of macrophage migration inhibitory factor in the bladder: evidence for a viscerovisceral reflex. AB - PURPOSE: Macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, is found in preformed stores in bladder epithelium. We examined the effects of prostatic inflammation on micturition frequency, bladder histology and bladder MIF content as a model in which to study viscerovisceral reflexes mediating pelvic visceral inflammation. MATERIALS AND METHODS: Cystometry was performed in urethane anesthetized male rats. Formalin or saline was injected into the ventral lobe of the prostate to induce inflammation. Cystometry continued 1 hour after injection. The bladder, ventral lobes of the prostates and lumbosacral spinal cord were then removed, and protein levels and gene expression of MIF, cyclooxygenase-2 (COX-2) and nerve growth factor (NGF) were examined. Edema was verified histologically in the bladder and prostate. RESULTS: Intraprostatic formalin produced almost immediate bladder hyperreflexia, which was maintained during the observation period. Bladder edema was noted during histological examination. Bladder MIF protein amounts decreased, while COX-2 and NGF increased after prostatic injection. Bladder MIF, COX-2 and NGF mRNA increased. In the lumbosacral spinal cord protein and mRNA amounts increased for all factors examined in animals that received intraprostatic formalin. No changes were observed in the cervical cord. Rats injected with formalin mixed with dye showed restriction of the dye to the prostate. CONCLUSIONS: A viscerovisceral reflex in the rat, probably mediated by the lumbosacral spinal cord, produced bladder hyperreflexia and bladder edema, and evoked MIF release from the bladder and the induction of other inflammatory mediators. This supports our hypothesis that MIF is involved in neurogenic inflammation in the pelvic viscera and it may represent an interesting therapeutic target. PMID- 15538288 TI - Evaluation of polyethylene glycol based hydrogel for tissue sealing after laparoscopic partial nephrectomy in a porcine model. AB - PURPOSE: Polyethylene glycol (PEG) based hydrogel is available as a tissue sealant and hemostatic aid. We determined the long-term safety and efficacy of its use as a tissue sealant for laparoscopic partial nephrectomy in a porcine model. MATERIALS AND METHODS: A total of 16 swine were cycled to 1 control group and 3 treatment groups, which underwent laparoscopic partial nephrectomy with hemostasis achieved only with application of a biodegradable PEG based hydrogel. The 3 treatment groups were sacrificed at 2, 6 and 12 weeks, respectively. Humoral immune response to the hydrogel used in the porcine abdomen was examined using enzyme-linked immunosorbent assay to detect antibodies in the serum at 0, 2, 6 and 12 weeks. Cell mediated immune response was examined using a lymphocyte proliferation assay to measure the response of leukocytes to various mitogens and antigens, including the polymerized hydrogel, at the same intervals. RESULTS: Hemostasis was satisfactory after hydrogel application. No adverse effects in the immediate and delayed periods were noted. At 2, 6 and 12 weeks there were no significant differences in hemoglobin or creatinine levels, or in the humoral immune response by enzyme-linked immunosorbent assay. There was no significant difference between test and control pig reactivity to hydrogel as an antigen in the lymphocyte proliferation assay at any time point. Histologically by 6 weeks the animals had almost absorbed the hydrogel with acute inflammation and foreign body reaction resolving by 6 to 12 weeks. No deleterious effect to renal tubules was seen. CONCLUSIONS: Biodegradable PEG based hydrogel is effective for long term use as an agent for hemostasis. There was no detectable humoral immune response and no cell mediated immune response to sealant after 2 weeks. This represents promising sealant technology and it should be further investigated for human use. PMID- 15538289 TI - Caveolae are negative regulators of transforming growth factor-beta1 signaling in ureteral smooth muscle cells. AB - PURPOSE: The mechanisms underlying ureteral cell regulation are largely unknown. Previous studies have identified lipid rafts/caveolae as regulators of growth stimulatory signals in ureteral smooth muscle cells (USMCs). In this study we determined whether growth inhibitory signaling by transforming growth factor beta1 (TGF-beta1) is also regulated by caveolae in USMC. MATERIALS AND METHODS: Expression of components of the TGF-beta1 signaling axis in USMCs was determined by immunoblot and mRNA analyses. Growth regulatory activity of TGF-beta1 was assessed by H-thymidine incorporation. In select experiments caveolae were disrupted reversibly by cholesterol depletion and replenishment prior to TGF beta1 treatment. TGF-beta1-responsive gene expression was evaluated using the TGF beta1 responsive promoter-reporter construct 3TP-Lux. RESULTS: USMCs expressed TGF-beta1, types I and II TGF-beta1 receptors, and the effector Smad-2. TGF-beta1 potently inhibited DNA synthesis in USMCs (IC50 60 pM). TGF-beta1 mediated DNA synthesis inhibition was potentiated following the disruption of caveolae by cholesterol depletion. This effect was reversible with membrane cholesterol restoration. TGF-beta1 stimulated gene activity was augmented by caveolae disruption, while caveolae reformation returned promoter activity to baseline levels. CONCLUSIONS: TGF-beta1 is a potent growth inhibitor of USMCs and its activity can be enhanced by caveolae ablation. These findings suggest a role for TGF-beta1 in the growth regulation of normal ureteral cells and implicate caveolar membrane domains in the negative regulation of TGF-beta1 signaling. These studies may be relevant to ureteral pathologies that are characterized by smooth muscle dysplasia. PMID- 15538290 TI - Atorvastatin ameliorates renal tissue damage in unilateral ureteral obstruction. AB - PURPOSE: The current study was done to determine whether atorvastatin, the HMGCoA (3-hydroxy-3-methylglutaryl CoA) reductase inhibitor, could decrease renal transforming growth factor-beta (TGF-beta) levels in unilateral ureteral obstruction (UUO) and concomitantly affect renal tissue damage in UUO. MATERIALS AND METHODS: Atorvastatin (20 mg/kg) was administered to rats 1 day prior to UUO and every day thereafter. Kidneys were harvested at day 14 after UUO. Tissue TGF beta was measured by bioassay using mink lung epithelial cells. Renal tubular proliferation and apoptosis were detected by immunostaining proliferating cell nuclear antigen and polyclonal antisingle strand DNA antibody, respectively. Fibrosis was assessed by measuring collagen deposition with trichrome stained slides. Interstitial leukocyte was detected by immunostaining CD45. RESULTS: TGF beta bioassay showed that the obstructed kidney in the control group contained significantly higher TGF-beta than the unobstructed kidney in the control group (mean +/- SD 79.1 +/- 48.5 vs 28.7 +/- 13.7 pg/mg tissue) and atorvastatin significantly decrease tissue TGF-beta in the obstructed kidney (53.4 +/- 37.0 pg/mg tissue). Immunostaining polyclonal antisingle strand DNA antibody demonstrated that the obstructed kidney in the control group has significantly more tubular apoptosis than the unobstructed counterpart (4.8 +/- 2.8 vs 2.1 +/- 1.2 nuclei per high power field) and atorvastatin significantly decreased renal tubular apoptosis in the obstructed kidney (1.1 +/- 0.7 nuclei per high power field). In addition, immunostaining proliferating cell nuclear antigen showed that the obstructed kidney in the atorvastatin group had significantly more renal tubular proliferation than the obstructed kidney in the control group (48.7 +/- 20.8 vs 17.3 +/- 10.6 per high power field). Control obstructed kidney showed significantly more fibrosis, which was also blunted by atorvastatin. CONCLUSIONS: Atorvastatin significantly decreases tissue TGF-beta, resulting in a decrease in tubular damage and interstitial fibrosis. This suggests that atorvastatin is a promising agent for preventing renal tubular damage in UUO. PMID- 15538291 TI - Cystometric findings in mice lacking muscarinic M2 or M3 receptors. AB - PURPOSE: The physiological importance of muscarinic M3 and M2 receptors for bladder function was investigated in vivo using mice lacking M3 or M2 receptors and littermate WT controls. MATERIALS AND METHODS: Unanesthetized mice of each sex underwent continuous cystometry before and after administration of atropine (1 mg/kg). RESULTS: Male M3 knockout (KO) mice had longer voiding intervals, and larger micturition volumes and bladder capacity than M2 KO or WT males. There was no significant difference in any cystometric parameters between male M2 KO and WT mice. In females M3 KO and M2 KO mice had longer voiding intervals and larger micturition volumes than WT animals. Atropine had marked inhibitory effects on voiding efficacy in WT and M2 KO mice but it had no effect on any cystometric parameters in M3 KO mice. CONCLUSIONS: The current results confirm that M3 receptor is the principal muscarinic receptor subtype responsible for bladder contraction and the role of M2 receptors is of minor importance. Functional impairments found in M3 KO mice were milder than those elicited by acute blockade of muscarinic receptors by atropine in WT mice, suggesting that noncholinergic mechanisms can compensate for a chronic loss of M3 receptors. PMID- 15538292 TI - Effects of chronic administration of doxazosin on alpha1-adrenoceptors in the rat prostate. AB - PURPOSE: Although the clinical efficacy of alpha1-adrenoceptor (alpha1-AR) antagonists for the treatment of benign prostatic hyperplasia is not disputed, their mechanism of action and ability to maintain long-term effectiveness have only recently been investigated. Since it is known that chronic administration of receptor antagonists causes up-regulation in the targeted receptor, we examined the effects of chronic administration of doxazosin, a nonspecific long acting alpha1-AR antagonist, on the properties of alpha1-AR subtypes in the rat prostate. MATERIALS AND METHODS: Rats were treated with doxazosin (2 or 4 mg/kg daily subcutaneously, supplemented with 4 mg/kg daily orally) for 8 or 12 weeks. Prostatic alpha1-AR properties at the protein and gene transcript levels were quantified by radioligand receptor binding and real-time reverse transcriptase polymerase chain reaction, respectively. RESULTS: Treated rats that received the highest levels of doxazosin had significantly heavier prostates compared with age matched controls. After 12 weeks of treatment radioligand binding studies with radiolabeled alpha1-AR antagonist demonstrated no significant differences in the density of total alpha1-ARs in the prostate, whereas the results of real-time reverse transcriptase-polymerase chain reaction showed up-regulation in the mRNA expression levels of all 3 alpha1-AR subtypes (alpha1A, alpha1B and alpha1D) in the ventral and dorsolateral regions of the rat prostate. CONCLUSIONS: These data demonstrate that chronic treatment with doxazosin causes an alteration in the properties of the alpha1-AR system in the rat prostate. These findings may provide insight into the long-term effectiveness of alpha1-AR antagonists in the treatment of benign prostatic hyperplasia. PMID- 15538293 TI - Renal tolerance to prolonged warm ischemia time in a laparoscopic versus open surgery porcine model. AB - PURPOSE: To our knowledge the effects of renal warm ischemia (WI) during laparoscopic vs open surgery have not been investigated. Decreased renal blood flow during pneumoperitoneum may precondition the kidney to tolerate longer WI time. Traditionally 30 minutes has defined the limit of renal WI time in open surgery. However, recent reports show renal function recovery at WI times of 45 to 120 minutes. We assessed renal function recovery after prolonged WI during laparoscopic vs open surgery in a solitary kidney porcine model. MATERIALS AND METHODS: A total of 32 female farm pigs underwent right laparoscopic nephrectomy to create a single kidney model. At 12 days later the animals were randomized into open and laparoscopic groups, each with 4 subgroups, namely 30, 60 and 90 minutes of WI, and a 90-minute control. Serum creatinine and the glomerular filtration rate were assessed preoperatively and on postoperative days 1, 3, 8 and 15. RESULTS: There was no statistical difference in renal function between the laparoscopic and open groups. Although the early decrease in renal function (72 hours) was highest in the 60 and 90-minute WI groups, by postoperative day 15 this difference was not statistically significant. Postoperative day 15 glomerular filtration rate and serum creatinine values were not significantly different from baseline in any of the WI groups. CONCLUSIONS: We found no difference in renal function recovery when comparing laparoscopic and open WI. Although WI up to 90 minutes resulted in initial renal dysfunction, by 2 weeks postoperatively function normalized. Our results indicate that in a single kidney porcine model the renal unit can fully recover from WI times of up to 90 minutes. PMID- 15538294 TI - p53 expression and apoptosis in varicocele in the rat testis. AB - PURPOSE: We investigated the presence of p53 expression and apoptosis in an experimental model of varicocele in the rat. MATERIALS AND METHODS: A total of 30 adult male Sprague-Dawley rats were studied in 3 groups, namely group 1-13 with varicocele, group 2-9 with sham operation and group 3-8 controls. All animals underwent orchiectomy after 30 days and sections of the left testis from each animal were studied. TUNEL was used to detect apoptotic germ cells and an apoptotic index (AI) was calculated for each testis specimen. p53 expression was determined immunohistochemically and graded according to the proportion of cells stained in each specimen. RESULTS: AI values in group 1 testes were 0% to 0.51% (mean 0.06% +/- 0.13%) and the corresponding range in group 2 was 0.04% to 0.69% (mean 0.25 +/- 0.21%). No group 3 control specimens showed TUNEL staining (AI 0% in all). Although the mean AI in group 1 was higher than that in controls, this difference was not statistically significant (p = 0.318). The mean AI value in group 2 was significantly higher than in the control group (p = 0.021) but it was comparable to that in group 1 (p = 0.099). Seven (53.8%) and 2 (22.2%) specimens in groups 1 and 2, respectively, showed p53 staining in germ cell nuclei. None of the specimens in group 3 showed nuclear p53 expression. The grade of p53 expression in group 1 was significantly higher than that in groups 2 (p = 0.042) and 3 (p = 0.011). Expression grades in groups 2 and 3 were not significantly different (p = 0.426). CONCLUSIONS: To our knowledge p53 expression that may reflect abnormal spermatogenesis has been documented for the first time in an experimental model of varicocele in the rat testis. In contrast to other investigations, no link between apoptosis and varicocele was detected. PMID- 15538295 TI - Over expression of thioredoxin-1 in transgenic mice attenuates germ cell apoptosis induced by experimental cryptorchidism. AB - PURPOSE: Experimental cryptorchidism induces apoptosis in testicular germ cells by generating reactive oxygen species. We investigated the effects of a redox regulating molecule, thioredoxin-1 (TRX1), on testicular damage caused by experimental cryptorchidism. MATERIALS AND METHODS: Unilateral cryptorchidism was surgically induced in TRX1 transgenic (TRX1-Tg) or WT adult C57BL6 mice. The contralateral scrotal testis served as a control. RESULTS: Experimental cryptorchidism decreased testicular weight in WT mice from 4 days after surgery. The decrease in testicular weight was significantly attenuated in TRX1-Tg mice compared with WT mice 7 to 14 days after surgery (p <0.01). However, the difference between the 2 groups was not significant 28 days after surgery. Histological analysis and TUNEL assays demonstrated that apoptosis occurred in germ cells of the cryptorchid testis in each group but the appearance of apoptotic germ cells was delayed by 3 days in TRX1-Tg mice. CONCLUSIONS: TRX1 over expression suppressed apoptosis in testicular germ cells induced by experimental cryptorchidism, indicating that TRX1 intensification may be a useful therapeutic strategy for male infertility associated with heat stress. PMID- 15538296 TI - Re: inflammation as a target for prostate cancer chemoprevention: pathological and laboratory rationale. PMID- 15538299 TI - Re: nephron sparing surgery for appropriately selected renal cell carcinoma between 4 and 7 cm results in outcome similar to radical nephrectomy. PMID- 15538301 TI - Re: is antegrade stenting superior to retrograde stenting in laparoscopic pyeloplasty? PMID- 15538302 TI - Re: the economic burden of metastatic and prostate specific antigen progression in patients with prostate cancer: findings from a retrospective analysis of health plan data. PMID- 15538306 TI - [Neurobiology and pharmacotherapy of social phobia]. AB - Social phobia (also known as social anxiety disorder) is still not clearly understood. It was not established as an authentic psychiatric entity until the diagnostic nomenclature of the American Psychiatric Association DSM III in 1980. In recent years, increasing attention among researchers has contributed to provide important information about the genetic, familial and temperamental bases of social phobia and its neurochemical, neuroendocrinological and neuroanatomical substrates, which remain to be further investigated. Up to date, there have been several findings about the possible influence of variables, including particularly genetic, socio-familial and early temperamental (eg behavioral inhibition) factors that represent risk for the later development of social phobia. Clinical neurobiological studies, based on the use of exogenous compounds such as lactate, CO2, caffeine, epinephrine, flumazenil or cholecystokinin/pentagastrin to reproduce naturally occurring phobic anxiety, have shown that patients with social phobia appear to exhibit an intermediate sensitivity between patients with panic disorder and control subjects. No difference in the rate of panic attacks in response to lactate, low concentrations of CO2 (5%), epinephrine or flumazenil was observed between patients with social phobia and normal healthy subjects, both being less reactive compared to patients with panic disorder. However, patients with social phobia had similar anxiety reactions to high concentrations of CO2 (35%), caffeine or cholecystokinin/pentagastrin than those seen in patients with panic disorder, both being more intensive than in controls. Several lines of evidence suggest specific neurotransmitter system alterations in social phobia, especially with regard to the serotoninergic, noradrenergic and dopaminergic systems. Although no abnormality in platelet serotonin transporter density has been found, patients with social phobia appear to show an enhanced sensitivity of both post-synaptic 5HT1A and 5HT2 serotonin receptor subtypes, as reflected by increased anxiety and hormonal responses to serotoninergic probes. Platelet 5HT2 receptor density has also been reported to be positively correlated to symptom severity in patients with social phobia. During anticipation of public speaking, heart rate was elevated in patients with social phobia compared to controls. Norepinephrine response to the orthostatic challenge test or to the Valsalva maneuver was also greater in patients with social phobia. While normal beta-adrenergic receptor number was observed in lymphocytes, a blunted response of growth hormone to clonidine, an a2-adrenergic agonist, was reported. This suggests reduced post synaptic a2-adrenergic receptor functioning related to norepinephrine overactivity in social phobia. Decreased cerebrospinal fluid levels of the dopamine metabolite homovanillic acid have also been observed. There are relatively few reports of involvement of the adrenal and thyroid functions in social phobia, and all that has been noted is that patients with social phobia show an exaggerated adrenocortical response to a psychological stressor. Recent advances in neuro-imaging have contributed to find low striatal dopamine D2 receptor binding or low dopamine transporter site density in patients with social phobia. They have also demonstrated the involvement of the cortico-limbic pathways, including the prefrontal cortex, hippocampus and amygdala, which show an increased activity in different experimental conditions. These brain regions have extensively been reported to play an important role in the cognitive appraisal in determining the significance of environmental stimuli, in the emotional and mnemonic integration of information, and in the expression of contextual fear-conditioned behaviors, which might be disrupted in the light of the phenomelogical aspects of social phobia. A substantial body of literature based on case reports, open and placebo-controlled trials, has now clearly examined the efficacy of major classes of psychotropic agents including monoamine oxidase inhibitors, beta-blockers, selective serotonin reuptake inhibitors and benzodiazepines in social phobia. Until recently, irreversible non-selective monoamine oxidase inhibitors, of which phenelzine was the most extensively evaluated, were considered as the most efficacious treatment in reducing the symptomatology associated with social phobia in 50-70% of cases after 4 to 6 weeks. However, side effects and dietary restrictions limit their use. This led to the development of reversible inhibitors of monoamine oxidase A, for which careful dietary monitoring is not required. Moclobemide has been the most widely studied but produced unconvincingly therapeutic effects on social phobic symptoms. To date, selective serotonin reuptake inhibitors may be considered as a reasonable first-line pharmacotherapy for social phobia. There is growing evidence for the efficacy of the selective serotonin reuptake inhibitors fluvoxamine, fluoxetine, citalopram, paroxetine and sertraline. They have beneficial effects with response rates ranging from 50 to 80% in social phobia. It has been recommended that the treatment period should be extended at least 6 months beyond the early improvement achieved within the first 4 to 6 weeks. The overall advantages include tolerability with a low risk of adverse events. The benzodiazepines clonazepam and alprazolam have also been proposed for the treatment of social phobia. Symptomatic relief occurred in 40 to 80% of the cases with a relatively rapid onset of action within the first two weeks. Untoward effects, discontinuation-related withdrawal symptoms and abuse or dependence liability constitute major concerns about the use of benzodiazepines, so they should be reserved for cases unresponsive to the safer medications cited above. Beta-blockers such as atenolol and propanolol have commonly been employed in performance anxiety, decreasing autonomic symptoms (eg, tachycardia, sweating and dry mouth). However, they are not effective in the generalized form of social phobia. Other pharmacologic alternatives seem helpful for the management of social phobia, including venlafaxine, gabapentin, bupropion, nefazodone or augmentation with buspirone. Preliminary studies point to promising effects of these agents. Larger controlled clinical trials are now needed to confirm their potential role in the treatment of social phobia. PMID- 15538307 TI - [Diogenes syndrome: a transnosographic approach]. AB - Diogenes syndrome is a behavioural disorder of the elderly. Symptoms include living in extreme squalor, a neglected physical state and unhygienic conditions. This is accompanied by a self-imposed isolation, the refusal of external help and a tendency to accumulate heteroclite objects. This particular geriatric syndrome has been described for the first time only quite recently, as the 2 primary descriptions by geriatricians and psychiatrists date from 1966 and 1975 respectively. Its rare occurrence contrasts with the fact that it is well-known, partly due to it being named after the Greek philosopher "Diogene de Sinope", who taught cynicism philosophy and a return to a natural way of life, and partly because of its rare characteristics. The Diogenes syndrome is a fascinating object of study for the clinician who takes care of patients living in uncommon conditions, on the edge of society and unaware of the particularity of their lifestyles. Patients suffering from Diogenes syndrome are usually discovered by chance, either because of a somatic illness, or as a result of social intervention related to their behavioural problems. Management of the syndrome is difficult and ethically challenging, as the patient does not seek help. Moreover, 46% of patients have a 5 year mortality rate. Hospitalisation has to be avoided whenever possible and ambulatory treatment and social measures should be favoured. Psychotropic treatment prescription may be necessary, depending on clinical features and the possible underlying psychiatric disease. Although several clinical hypotheses have been suggested, the true ethiopathogeny of the syndrome remains unclear. Most authors agree that this behaviour does not reflect free will and has consequently no theoretical relationship to the Greek philosopher. There is no true consensus about diagnostic criteria. They include the main features of the syndrome and exclude known psychiatric syndromes. Clark and Mankikar, who named this syndrome, reckon it may represent stress-related defence mechanisms of the elderly or may be related to natural ageing process. However, psychiatric pathologies as paranoid and paranoiac psychoses, mood disorders and obsessive and compulsive disorders have been described to be associated with it in the literature. Dementia, in particular temporo-frontal dementia, should be looked for and excluded clinically. Alcohol abuse seems to be an aggravating rather than a precipitating factor. Finally, the link between these pathologies and Diogenes syndrome is not yet determined: are they triggering, co-morbid or etiological factors? Should this syndrome be considered as a true illness or as a symptom? This paper presents Diogenes syndrome as a behavioural disorder and distinguishes 2 types: the "active type"--patients who collect from outside to clutter inside--and the "passive type"--patients who passively become invaded by their rubbish. Active type patients fill their home to fill in the vacuum of their life, as it deteriorates and looses its narcissical appeal. Passive type patients accumulate by default and emptiness. A psychopathological understanding is presented here, referring to psychoanalytical theories of the Moi-peau (ego-skin) described by Anzieu. The Moi-peau represents a structure of the psyche founded on the following principle: any psychic function develops itself according to a bodily function from which it transposes its functioning at a mental level. The skin has three functions: the containing shell, the protective barrier of the psyche, and a medium of exchange. The Moi peau is organised as a double-wall acting both as a defence mechanism and as a filter between the psyche and the external world. It preserves the relationship and the cohesion "container-content". As a result of a narcissical wound, the Moi peau is damaged and looses its function of a container. In the case of Diogenes Syndrome, the accumulated items repair the Moi-peau and the home becomes an "exterior-proof", thus playing the role of the Moi-peau. This behaviour therefore plays a repairing role for psychic functioning, allowing psychic survival. PMID- 15538308 TI - [Posttraumatic stress state: a therapeutic lever]. AB - Chronic post-traumatic stress disorder (PTSD) is a very complex syndrome which is hard to detect because of the multiplicity of its expressions. Further more, these clinical expressions are far from the "pure" syndrome described in the DSM IV. So, the clinician faces a dilemma: how can he account for the traumatic clues without using the PTSD as a ragbag of a diagnosis? We found the way to discard this dilemma when we decided to use what M. Struber said about her experience with cancer and PTSD. She suggests not to emphasize psychopathology and to use a post-traumatic stress framework. This way to reframe some psychiatric urgencies is very useful because it gives back ability to the patient. When using a post traumatic stress framework we tell the patient and his family that we acknowledge he has defensible reasons for not managing with an event which, we acknowledge too, was traumatic for him. In that way we begin to explore what each person is experiencing, because the traumatic experiencing is generally shared by the patient and his family. The members of the family are often angry and fed up of the patient behaviour and think themselves as victims of him. On the other part, the patient feels himself as a misunderstood person, victim of the others. The primary trauma is forgotten for a long time or nobody make any link between it and what is happening in the present. The manifestations of the PTSD initiate subsequent aftermaths and suffering for everybody. When working with psychiatric emergencies, we have to manage with acute situations in which each people is both victim and aggressor and in which clinicians run the risk of being given the role of either victim or aggressor. The trial of strength played between the patient and his family is going to be played with the clinician. These situations are described by S. Lamarre when she speaks of "victimisation" and are overloaded with control stake. Each one tries to make the other guilty and disgraced, and the clinician is at risk to feel and/or make feel in the same way the patient and his family. These situations are blocked and the temptation is to resort to a kind of coup when the clinician decides it's enough! and forces his opinion and decision. What is not a very good way to create the essential therapeutic co operation! In this article we show how using a post-traumatic stress framework is very useful to reframe the situation of "victimisation", give the opportunity to discard its trap, open a new sight which allows to find new solutions and promote a therapeutic co-operation. It's important to stress the fact that it's not efficient to use a post-traumatic stress framework as a formula. The clinician who uses it has to feel it, otherwise he will be unable to co-create this new reality with the system he entered, when receiving the emergency. PMID- 15538309 TI - [Validation of the French Version of the Life Skills Profile with people suffering of schizophrenia]. AB - INTRODUCTION: Since the discovery of neuroleptics and the reintegration of people with schizophrenia in the community, psycho-social reeducation became an essential part of the treatment. The Life Skills Profile is a tool of reference for assessment of the dimensions, which have an impact on the adaptation in community. Each item describes an observable behavior, written in common language, to allow the care-givers and the family to evaluate it without having a specific formation. The long version (39 items) is recommended for therapeutic interventions with a person and the short one (20 items) for large scale studies on outcome in community. AIM OF THE STUDY: In order to dispose of a measure of function and disability in schizophrenia, we have translated the Life Skills Profile (LSP) in French and tested the validity of this translation for the long version (39 items) and the short ones (16 and 20 items). The 4 dimensions of the 16-items version--"withdrawal", "self-care", "compliance" and "antisocial"--were used for people with mental disorders and the 20-items version enhanced with a fifth dimension "bizarre", especially useful for people with schizophrenia. METHOD: 175 people suffering of schizophrenia (DSM IV, codes F20.0 to F20.5) were evaluated by their caregivers in 3 settings: psychiatric hospital, ambulatory care and sheltered homes. Confirmatory factorial analyses were performed to test the dimensional models and their psychometric characteristics were established. RESULTS: The original structure in 5 dimensions of the long version (39 items) is not confirmed. However, the short versions in 16 and 20 items were confirmed, without any modification. The required psychometric qualities of reliability and validity of the 20-items version were fulfilled. The inter-rater reliability ranged from 0.65 to 0.75 for the 5 dimensions; the test-retest reliability ranged from 0.80 to 0.91 and the internal consistency from 0.67 to 0.81. The validity was evaluated by comparison of the LSP scores upon living arrangements (people living in sheltered homes had lesser scores) and pathologies (19 people with severe major depression had higher scores). CONCLUSION: The 39-items version of the Life Skills Profile was not validated in French. However, for clinical practice of social rehabilitation, this tool remains useful for a single person to check specific behaviors, which could hinder his/her integration into the community, to plan specific interventions and to evaluate changes, in addition with other scales. By the validation of the short version of the Life Skills Profile, an instrument is at disposal in French for outcome studies which allow to: 1) quickly assess the social functioning of person suffering of schizophrenia by a caregiver or a family member; 2) to detect insufficient skills in specific domains; and 3) to evaluate therapeutic efficiency. PMID- 15538310 TI - [Sleep in adolescents and its disorders. A survey in schools]. AB - OBJECTIVES: Numerous studies emphasized the high prevalence of sleep problems in adolescents. However, it remains to be seen whether these problems are developmental or if they are related to pathological conditions. In order to try to answer this question, we conducted an epidemiological study investigating the prevalence and correlates of sleep disorders in a population of high school adolescents. METHOD: The sample for this investigation consisted of 652 high school students (344 males and 308 females), aged 13 to 19 years (mean age: 15.1 1.2 years), attending colleges in the North of France. Data for the study were obtained by means of revised versions of the INSERM self-report questionnaires for adolescents. One questionnaire included 113 response choice items investigating the sleep parameters and the sociodemographic, behavioural, health, and environmental condition of the subject. This questionnaire was completed by a self-portrait questionnaire including 27 response choice items about mood, anxiety, instability and the quality of relations with others. In addition, a clinical examination was performed by the clinicians of the school health department in order to investigate the medical history of the subject and his (her) pubertal development. RESULTS: In a first step, statistical analysis showed some significant differences in sleep rates and troubles between males and females. If the total sleep time was found similar in males and females, females were found exhibiting significantly earlier bedtime (p<0.01) and waking time (p<0.005) than males. In addition, trouble falling asleep (p<0.0001), nightmares (p<0.001), need for daytime sleep (p<0.05), and stimulant pill intake (p<0.05) were found significantly more frequent in females. Then, the adolescents were classified into two subgroups. "Insomnia group" included the students who answered "often" or "always" to one of the five questions about: having trouble falling asleep, the occurrence of early awakenings, their need for daytime sleep, sleeping pill intake, and bad sleep quality. "Non insomnia group" included those who answered "never" or "sometimes" in response to the five questions. Results showed that 233 adolescents (35.7%) exhibited persistent sleep disorders, insomnia type. 40.2% of females and 31.6% of males were included in the "insomnia group" (p<0.05). Pubertal development was found significantly more advanced in the "insomnia group" adolescents, more particularly in females. In the same way, concerns about body weight (46.7% vs 38.8%; p<0.005), size and shape (15% vs 8%; p<0.01) were found significantly more frequent in the "insomnia group"; 32.3% of the "insomnia group" adolescents vs 17.9% in the "non insomnia group" reported health problems (p<0.0001). School difficulties were found significantly more frequent in the "insomnia group" compared to the "non insomnia group". The proportion of subjects who have previously repeated at least one school year (p<0.01), who reported bad school performances (p<0.01), and who reported to be "sometimes" or "often" away from school (p<0.01) was significantly higher in the "insomnia group" than in the "non insomnia group". In the same way, data showed that school investing was also significantly weaker in the "insomnia group" adolescents (p<0.01). The proportion of subjects who believed that their relations with parents (12.4% vs 6%; p<0.05), teachers (21.4% vs 10.5%; p<0.0001), or peers (10.2% vs 4.1%; p<0.002) was found significantly higher in the "insomnia group". Risk behaviours were also significantly more frequent in the "insomnia group" adolescents: suicidal ideation (25.1% vs 11.7%; p<0.001), suicide attempt (7.3% vs 3.6%; p<0.05), psychotropic drug use (11% vs 4.9%; p<0.02), regular stimulant drug use (10% vs 5.1%; p<0.005), regular tobacco use (7.8% vs 2.2%; p<0.001), regular alcohol use (13.3% vs 8.3%; p<0.05), and illicit drug use (5.2% vs 1%; p<0.001). Lastly, all the dimensions of the self-portrait were found significantly more affected or negative in the "insomnia group" than in the "non insomnia group". For example, the proportion of subjects who described themself usually sad (10.2% vs 3.4%; p<0.001), anxious (54.9% vs 40.5%; p<0.0001), downcast (41.1% vs 32.6%; p<0.05), or instable (56.9% vs 41.7%; p<0.0001) was significantly higher in the "insomnia group". CONCLUSION: In this descriptive study, if some data suggest that pubertal development may be involved in the sleep rates or troubles observed in adolescents, the results also show that persistent sleep disorders are significantly associated with physical, psychological or social difficulties. In summary, this study shows that it is essential to take an interest in the quality of sleep in adolescents, which may be a way to approach their psychosocial difficulties. PMID- 15538311 TI - [Is there an interaction between sleep-disordered breathing, depression and apolipoprotein E phenotype?]. AB - Sleep-disordered breathing (SDB) is widely underdiagnosed among adults. However, SDB may be considered as a public health problem because of clinical consequences for the patient: impaired awake performance, increased risk factor for cardiovascular diseases and increased prevalence of depression. Apolipoprotein E (apoE), a protein involved in lipid metabolism, has 3 major alleles e2, e3 and e4. Recently, it has been shown that apoE e4 allele, a well-known risk factor for cardiovascular diseases, was also associated with SDB. In this study, we assessed a potential interaction between SDB, depression and apoE phenotype. 92 male patients (36-79 years old, mean age 58.0 11.2) consulting in hospital for SDB were enrolled in the study. Each patient had the following exams: 1) overnight polysomnography to determine apnea/hypopnea index (AHI=average number of respiratory events 10 seconds with no breathing per hour). A moderate-to-severe SDB was defined with AHI 15. 2) a psychiatric examination to look for previous or present symptoms of depressive illness. 3) blood sampling to determine apoE genotype (using PCR-RFLP method). In our study, allele frequencies for apoE e2, e3 and e4 were similar to those reported in general population. Among 92 patients, 68 (74%) presented moderate-to-severe SDB and 28 (30%) previous or present symptoms of depressive illness. Our results indicate that: 1) apoE e4 was significantly associated with moderate-to-severe SDB (n=92, p=0.03), 2) scores of apnea-hypopnea index were significantly higher in e4-positive versus e4-negative participants (n=57, p=0,05) and 3) ApoE and depression were not linked. This study confirms a potential interaction between SDB and apoE phenotype, as recently reported. This suggests that e4 allele might be a genetic risk factor for SDB (e4 allele frequency higher in patients with moderate-to-severe SDB versus general population) and/or consequently a deleterious factor for this pathology (increased AHI in e4-positive versus e4-negative patients). Depression might be only one of clinical consequences of SDB. Thus, SDB leads to repeated hypoxemia and numerous awakenings resulting in fatigue and decreased cognitive abilities suitable to the onset of depressive illness in vulnerable persons. PMID- 15538312 TI - [Transcranial magnetic stimulation in cognition and neuropsychology]. AB - Classical neuropsychology relies on patients with irreversible brain lesions and cognitive impairments give informations about normal brain function. Transcranial Magnetic Stimulation (TMS) is a non-invasive method which involves placing an electromagnetic coil on the scalp. A pulse generates a magnetic field and this one passes, unattenuated by the skin and scalp, into the cortex inducing a current which results in neural activity. The technique shows a good temporal resolution and, moreover, because it represents an interference technique, can be said to have excellent functional resolution. For this reason, TMS appears to be a new tool for research in neuropsychology, producing transitory 'virtual lesion'effects which could help to understand how, when and where cognitive tasks are performed. The purpose of this article is to review recent research using TMS in cognition and neuropsychology, in a non exhaustive way. In safety studies, single TMS over motor cortex can produce simple movements. Several groups have applied TMS to the study of visual processing and found an impaired detection of visual stimuli. In a same way, TMS can disrupt speech when it was delivered in the language dominant hemisphere. Studies on the memory effects of TMS have been conflicting and the results seem to depend on the choice of paradigm and parameters. Other study depicted improvements in executive functioning after TMS on the left middle frontal gyrus or a diminution in reaction time during an analogic reasoning task. Moreover, some facial emotions seem to be less recognizable after TMS. Although TMS seem to be a new tool for neuro psychological investigations in healthy subjects, few studies reported cognitive effects of rTMS treatment in psychiatry. In a therapeutic view, many of these trials have supported a significant effect of TMS, but in some studies the effect is small and short lived. Several groups have reported on the use of rTMS as a treatment in resistant major depression and the impact on cognition functioning. Most of results tend to find no adverse cognitive effects after several weeks of daily rTMS in depressed patients, compared to Electroconvulsivo-therapy (ECT). The effects of transcranial magnetic stimulation (TMS) on hallucination severity and neurocognition were studied in a recent study. A statistically significant improvement was observed on a hallucination scale and on one cognitive measure. TMS is a promising tool for cognitive neuroscience and can provide complementary information to the one obtained using neuropsychological tests, and the one obtained using functional imaging techniques, which have superior spatial but inferior temporal resolution. PMID- 15538313 TI - [A study of the Edinburgh Postnatal Depression Scale (EPDS) on 859 mothers: detection of mothers at risk for postpartum depression]. AB - The postpartum is a high-risk period for the occurrence of anxious and depressive episodes. Indeed, during the first few days after delivery, mothers can present postpartum blues symptomatology: fatigue, anxiety, disordered sleeping and a changing mood. Postpartum depression is characterised by a changing mood, anxiety, irritability, depression, panic and obsessional phenomena. It occurs in approximately 10 to 20% mothers. The exact prevalence depending on the criteria used for detection. The first symptoms usually appear between the fourth and sixth week postpartum. However, postpartum depression can start from the moment of birth, or may result from depression evolving continuously since pregnancy. We can add that the intensity of postpartum blues is a risk factor that can perturb maternal development. So it is important for health professionals to dispose of predictive tools. This study is a validation of the French version of the EPDS. The aims of the study were to evaluate the postpartum depression predictive value at 3 days postpartum and to determine a cut-off score for major depression. Subjects participating in this study were met in 3 obstetrical clinics in, or in the vicinity of, Toulouse. Mothers with psychological problems, under treatment for psychological problems or mothers whose babies present serious health problems were excluded from the study. The EPDS was presented to 859 mothers (mean age=30.3; SD=4.5) met at one of the clinics at 3 days postpartum (period 1). They had an EPDS mean score of 6.4 (SD=4.6); 258 (30%) mothers had an EPDS score 9. 82.6% of these mothers experienced a natural childbirth and 17.3% a caesarean section; 51.5% gave birth to their first child, 36.2% to their second child and 12.3% to their third or more. All subjects were given a second EPDS with written instructions to complete the scale during the period 4 to 6 weeks postpartum and return it for analysis (period 2). Between the 4 to 6 weeks postpartum period, 722 mothers replied again to the EPDS. 131 mothers had an EPDS score 11 (mean age=30.3; SD=4.8). They had an EPDS mean score of 13.6 (SD=3.3). Mothers with probable depression were interviewed and assessed, using the Mini (Mini Neuropsychiatric Interview, Lecrubier et al. 1997), the SIGH-D (Structured Interview Guide for the Hamilton Depression Scale) and the BDI (Beck Depression Inventory) in order to diagnose a major depressive episode. They had a HDRS mean score of 13.7 (SD=5.1) and a BDI mean score of 13.6 (SD=5). At 3 days postpartum, we observed that 258 mothers (30%) had an EPDS scores 9 and 164 mothers (19%) had an EPDS scores 11. Between 4 and 6 weeks postpartum, we observed 18.1% of postpartum depression (EPDS 11) and 16.8% (EPDS 12) of major postpartum depression. The analysis of the sensitivity and the specificity at 3 days postpartum provides a cut-off score of 9 (Sensibility: 0.88) (Specificity: 0.50) as predictive of postpartum depression, for this cut-off score, the type I error is low (5.8%) but the type II error is more higher (18.9%). The analysis of the sensitivity and the specificity between 4 and 6 weeks postpartum provides a cut off score of 12 (Sensibility: 0.91) (Sensibility: 0.74) for the detection of major postpartum depression. Factor analysis shows at 3 days postpartum that the internal structure of the scale is composed of two subscales. The first factor F1 "anxiety" accounts 28% of the variance and the second factor F2 "depression" accounts 20% of the variance. Between 4 and 6 weeks postpartum, factor analysis suggests an unidimensional model in the evaluation of postpartum depression which is better than a two factor model. This factor accounts 40% of the variance. The scale has a good predictive value, and we can observe a significant correlation with the EPDS periods 1 and 2 (r=0.56; p<0.05). This result shows that the depressive mothers mood intensity predicts a future depressive risk. Furthermore, correlations between EPDS and BDI (r=0.68; p<0.05) and EPDS and HDRS (r=0.67; p<0.05) show a good convergent validity. The reliability study confirms the good internal consistency of the EPDS, at 3 days postpartum and in the postpartum depression -symptomatology evaluation (Cronbach's Alpha>0.80). In conclusion, this scale demonstrates good validity and is fast and easy use in obstetrical services, allowing early detection of women who risk to develop postpartum depression and, in the first week of postpartum, of mothers who suffer from a major postpartum depression. The use of the EPDS for an early screening of the risk of postnatal depression which is essential considering the consequences that postnatal depression can have on the development of the infant, on the quality of the relationship within the couple and on other social relationships. Mothers at risk for postnatal depression should be controlled and surveyed by the health professionals in obstetrical clinics. PMID- 15538314 TI - [Schizophrenia, diabetes mellitus and antipsychotics]. AB - During the last years, a contribution of antipsychotic drugs in the increase of diabetes prevalence in schizophrenic population has been repetitively suggested. The debate focused mainly on the second-generation antipsychotics. The analysis of the scientific literature indicates however that this discussion is not recent and an increase of diabetes prevalence in schizophrenic populations was already described before the introduction of neuroleptics. Then, after the introduction of the first neuroleptics in the 1950s, an increase of diabetes prevalence was reported among treated patients and the same alarms occurred in the 1990s after the introduction of second-generation antipsychotics. These treatments were related to an increase of glucose tolerance impairment, type II diabetes and diabetic acidoketosis. Recent epidemiological studies have confirmed the increase prevalence of diabetes in schizophrenic patients, particularly in schizophrenic patients before any antipsychotic treatment. Among the suggested mechanisms, there are sedentary life (due to hospitalisation and sedative effects of neuroleptics), food imbalance, shared genetic factors for diabetes and schizophrenia. Moreover, the frequency of the metabolic syndrome is increased in schizophrenic populations. This syndrome associates blood glucose increase, lipid metabolism disorders and android obesity. This could explain--via an increase of the cortisol production--the increase of mortality due to cardiovascular diseases observed in schizoprhenic patients. Thus, it seems well established that schizophrenia is associated with an increased risk for diabetes. It is however more difficult to evaluate the role of antipsychotic treatment as a causative factor of diabetes. Indeed, there are many published case reports or diabetes or diabetic acidoketosis after an antipsychotic treatment, but the level of evidence in controlled trials is low. Many studies were performed on large databases, but were retrospective and subjected to many flaws: concomitant diseases not taken into account, diabetes status evaluated by drug consumption, unknown diabetes status before antipsychotic treatment, etc. In the few prospective studies performed, no significant differences between the atypical versus typical antipsychotics were evidenced for new cases of diabetes. Moreover, in general population, the glucose tolerance impairment is underdiagnosed and it is estimated that people with a glucose tolerance impairment have a 5-10% annual risk of type II diabetes. Thus, this concern has to be replaced among the world epidemic increase of diabetes and in a population of patients whose the disease itself and life style are risk factors for diabetes. Some studies have explored the pathophysiological mechanisms that could support a diabetogenic effect of antipsychotics. Although it does not seem to be a direct effect of antipsychotics on insulin secretion by pancreatic cells, body weight increase has been evidence for both typical and atypical antipsychotics. However, it remains unclear whether this weight increase is responsible for a visceral adiposity, which is a risk factor better fitted to the cardiovascular mortality tha the body weight itself. Other hypotheses involving an effect on the leptin, which regulates the appetite, have been proposed. In waiting of new prospective controlled studies, and without denying the impact of antipsychotics on the glucose and lipid metabolisms (on the weight increase, for example), it should be recognized that the benefit/risk ratio remains largely in favour of the treatment, particularly for the atypical antipsychotics, more effective and better tolerated at the neurological level than the conventional antipsychotics. One of the benefits of the mainly articles in professional media about this concern is to draw attention on the metabolism disorders in schizophrenic patients, which are important risk factor of their frequent cardiovascular surmortality whatever the causes. Consequently, it is advised to monitor glucose and lipid metabolisms of schizophrenic patients before and during their treatment (body weight, fast blood glucose, blood cholesterol and triglycerides). In conclusion, schizophrenic patients are a population with an increased metabolic risk, which is a cause of their increased mortality. Although these data are known since a long time ago, this population does not benefit from the same metabolic follow-up than the non-schizophrenic population. The debate on the possible relationship between diabetes and antipsychotics should be also taken as a helpful recall of the necessity to follow simple rules of prevention and monitoring in this at-risk population. This should make it possible to preserve the benefit of the antipsychotics, the contribution of which in the treatment of schizophrenia is not any more to demonstrate. PMID- 15538315 TI - [Valpromide, Valproic acid and removal of small intestine in the treatment of a chronic depression: a case report]. AB - Valpromide (VPD) is an antiepileptic drug, derivative of Valproic acid (VPA), used as a mood-stabilizer in bipolar disorder for 25 years in several European countries. VPD is also used as an augmentation strategy in refractory depression. Despite chemical similarity between VPA and VPD, the pharmacokinetics of the 2 drugs in humans are quite distinct. We report a case of a patient, suffering from a bipolar treatment resistant depression, who dramatically improved after substituting VPD to VPA in association with fluoxetine. Mme X, 68 years old, has been hospitalized in March 2001 for the treatment of a resistant depression (TRD). She was suffering from removal of small intestine with chronic diarrhoea after a suicidal attempt two years ago. She had a bipolar disorder treated with VPD (1,200 mg/d) since 1 year. She presented a major depressive episode according to DSM IV with various symptoms like depressed mood, hypersomnia and difficulty initiating sleep, diminished ability to concentrate and to think, markedly diminished pleasure in all activities and major anxiety. Mme X fulfilled TRD diagnosis after resistance to two adequate antidepressants trials from different classes (clomipramine 175 mg/day and venlafaxine 300 mg/day). The antidepressant treatment (venlafaxine) was interrupted and she has been receiving a SSRI (fluoxetine 20 mg/day) for 4 weeks. After four weeks, she had a partial remission with persistent sleep problems, mood lability and anxiety. The VPA blood concentration was very low: 27 mg/L (normal range: 50 to 100 mg/L) in spite of a high dosage: 1,200 mg/day. Pharmacokinetic analysis of VPD shown that VPD transformation to VPA usually done in the intestine, was reduced because of the removal of hail intestine. We substituted VPD by VPA. Valproate blood concentration returned to normal range, induced dramatic improvement of depression within three days. VPD is an amide derivative of valproic acid (valproate), biotransformed by hydrolysis to its corresponding valproic acid. VPD is a prodrug of VPA. VPD is absorbed after transformation in gastro-intestinal mucous membrane. The adequate dosage of VPD (Depamide, 300 mg) is 4 to 6 tablets in acute manic phases, 2 to 4 tablets in long term treatment, 1 to 3 tablets in depressive episode. The biodisponibility of VPD is around 100% 75 and 90% of VPD is linked with protein albumin. The daily dosage determined the blood concentration of the active form (VPA), but this relation isn't linear. The optimal blood concentration of VPA (Depakine) ranges between 50 and 100 mg/L. the free form of VPA is influenced by protein disorders such as of hypoalbuminemia and by presence of fat acids in food. This case report demonstrates at a clinical level that VPD and VPA are not equivalent for treating bipolar depression. This case also suggests that a deep investigation of the pharmacokinetic of psychotropic drugs can help clinicians to resolve clinical problems of treatment of depression. PMID- 15538316 TI - [Persistent psychotic disorder following bilateral mesencephalo-thalamic ischaemia: case report]. AB - A 38-year old male patient with no history of psychiatric illness developed a progressive psychotic disorder after bilateral (predominantly left) mesencephalo thalamic cerebral ischaemia. The reason of the emergency hospitalization was the sudden onset of a confusional state, culminating in a fluctuating comatose status. The neurological examination found mild right hemiparesia, praxic disorders and reactive left mydriasis with paresia of the downward vertical stare, leading to the hospitalisation in the neurology department for suspicion of a cerebral vascular ischaemic accident. The psychiatric symptoms started with acoustic-verbal hallucinations, poorly structured paranoid delusions, progressively developed over two weeks, followed by behavioural disorders with psychomotor agitation and heteroaggressivity. The patient was transferred to the psychiatric department, because of the heteroaggressive risk and lack of morbid consciousness, in spite of recovering from the confusional status. An intensive psychiatric management was proposed, combining a psychotherapeutic approach with 4 mg of risperidone and adjustable doses of benzodiazepine according to the psychomotor agitation. During the next days, there was a net recovery of the behavioural disorders, in spite of the persistence of the ideas of persecution. All the neurological symptoms also decreased. An anomaly of the polygon of Willis was found on a cerebral arteriography (the posterior cerebral arteries had a foetal origin, dependent on carotidal axes and not on the vertebro-basilar system). The main emboligen risk factor was the presence of a permeable foramen ovale, discovered during a transoesophageal echography. The patient underwent a surgical correction of the permeable foramen ovale. The psychiatric hospitalization for three months was continued by ambulatory follow-up. The initial positive symptoms (delusions, acoustic-verbal hallucinations) progressively diminished while negative symptoms became predominant after few months. One year after the first hospitalization the patient presented a second psychotic decompensation, with delusions of persecution, jealousy and behavioural disorders with heteroaggressivity, that required an emergency psychiatric hospitalization. The wife of the patient decided to divorce, because she was "frightened by the threats of death" from her husband. A neurological assessment during the second hospitalization in psychiatry did not find new ischaemic lesions after the cardiac surgery for the permeable foramen ovale, nor relevant changes in cerebral perfusion. The final diagnosis after the psychiatric follow up of 14 months was: psychotic disorder with delusional ideas, due to cerebral ischaemia (DSM IV). There are relatively few data in the literature regarding persistent psychotic disorders in the context of ischaemic mesencephalo-thalamic lesions. However, several authors support the hypothesis of a possible disconnection of the thalamic nuclei, the frontal lobes and limbic system as a risk factor or a triggering factor for psychotic disorders in cerebral ischaemia. Observations concerning the occurrence of psychotic disorders following cerebral- especially localised--ischaemia may help to better understand the neuro physiological mechanisms triggering or accompanying the psychiatric symptomatology. The role of functional cerebral imagery in the detection of possible structural lesions related to clinical observations must be emphasised. The slow progression (over a year) to psychotic disorder with predominantly negative symptoms emphasized the importance for long-term prospective studies. Isolated clinical observations arouse the interest for a specific scale for psychotic disorders occurring after cerebral ischaemia, similar to existing specific scale for post-ischaemic depressive disorders. The necessity of systematic psychiatric examination and re-evaluations in thalamic cerebral ischaemia is to be highlighted, both for the differential diagnosis and for the early psychiatric care. PMID- 15538317 TI - State-specific prevalence of current cigarette smoking among adults--United States, 2003. AB - Cigarette smoking causes approximately 440,000 deaths annually in the United States. To assess the prevalence of current cigarette smoking among adults, CDC analyzed data from the 2003 Behavioral Risk Factor Surveillance System (BRFSS) survey. This report summarizes the results of that analysis, which indicated substantial variation in cigarette smoking prevalence in the 50 states, the District of Columbia (DC), Guam, Puerto Rico, and the U.S. Virgin Islands (USVI) (range: 10.0%-34.0%). To further reduce the prevalence of smoking, states/areas should implement comprehensive tobacco-control programs. PMID- 15538318 TI - Indoor air quality in hospitality venues before and after implementation of a clean indoor air law--Western New York, 2003. AB - Secondhand smoke (SHS) contains more than 50 carcinogens. SHS exposure is responsible for an estimated 3,000 lung cancer deaths and more than 35,000 coronary heart disease deaths among never smokers in the United States each year, and for lower respiratory infections, asthma, sudden infant death syndrome, and chronic ear infections among children. Even short-term exposures to SHS, such as those that might be experienced by a patron in a restaurant or bar that allows smoking, can increase the risk of experiencing an acute cardiovascular event. Although population-based data indicate declining SHS exposure in the United States over time, SHS exposure remains a common but preventable public health hazard. Policies requiring smoke-free environments are the most effective method of reducing SHS exposure. Effective July 24, 2003, New York implemented a comprehensive state law requiring almost all indoor workplaces and public places (e.g., restaurants, bars, and other hospitality venues) to be smoke-free. This report describes an assessment of changes in indoor air quality that occurred in 20 hospitality venues in western New York where smoking or indirect SHS exposure from an adjoining room was observed at baseline. The findings indicate that, on average, levels of respirable suspended particles (RSPs), an accepted marker for SHS levels, decreased 84% in these venues after the law took effect. Comprehensive clean indoor air policies can rapidly and effectively reduce SHS exposure in hospitality venues. PMID- 15538319 TI - Vaccination coverage among children entering school--United States, 2003-04 school year. AB - One of the national health objectives for 2010 is to sustain > or =95% vaccination coverage among children in kindergarten through first grade (objective 14-23). To determine the percentage of vaccination coverage among children entering kindergarten, data on vaccination coverage were analyzed from reports submitted to the National Immunization Program by states, the District of Columbia (DC), and eight current or former U.S. territories for the 2003-04 school year. This report summarizes the results of that analysis, which determined that coverage for all vaccines except hepatitis B (HepB) and varicella was reported at >90% in 45 areas. However, the vaccines required in each reporting area and the methods for surveying kindergarten-aged children vary substantially; in seven states, <20% of eligible children were surveyed. The wide variations in survey populations underscore the need for CDC to continue working with immunization programs in states, DC, and current or former territories to improve survey methods and automate reporting of data. PMID- 15538320 TI - Awareness of family health history as a risk factor for disease--United States, 2004. AB - Persons who have close relatives with certain diseases (e.g., heart disease, diabetes, and osteoporosis) are more likely to develop those diseases themselves. Family health history is an important risk factor that reflects inherited genetic susceptibility, shared environment, and common behaviors. Although clinicians are trained to collect family histories, substantial barriers exist to obtaining this information in primary care practice (e.g., lack of time or lack of reimbursement). To promote the use of family history as a screening tool for disease prevention and health promotion, several initiatives have called for new self-administered family history collection tools and educational programs to help clinicians interpret and apply family history information to patient care. To assess attitudes, knowledge, and practices of U.S. residents regarding their family health histories, CDC analyzed data from the 2004 HealthStyles Survey. This report summarizes the results of that analysis, which indicated that although 96.3% of survey respondents believe their family history is important for their own health, few have actively collected health information from their relatives to develop a family history. Targeted public health efforts are needed to 1) help persons collect family history information to share with their health care providers and 2) educate and assist providers to interpret and apply this information effectively. PMID- 15538321 TI - Preventive-care practices among adults with diabetes--Puerto Rico, 2000-2002. AB - Preventive-care practices among persons with diabetes can prevent or delay complications such as eye disease, kidney disease, or nerve damage that is a precursor to disabling foot disease. However, the level of diabetes-related preventive care is inadequate in the United States, and little has been reported about preventive care in Puerto Rico, where an estimated 10% of adults have diagnosed diabetes. CDC analyzed data from 2000, 2001, and 2002 Behavioral Risk Factor Surveillance System (BRFSS) surveys to assess the percentage of adults with diabetes in Puerto Rico who engaged in five selected preventive-care practices. This report summarizes the results of that analysis, which indicated that, with the exception of hemoglobin A1c testing, the percentages of adults engaging in preventive-care practices were lower than the target percentages set by U.S. national health objectives for 2010. PMID- 15538322 TI - West Nile virus activity--United States, November 3-8, 2004. AB - During November 3-8, a total of 41 cases of human West Nile virus (WNV) illness were reported from seven states (California, Maryland, Minnesota, New Mexico, Oregon, Tennessee, and Texas). PMID- 15538323 TI - The stem-cell state. PMID- 15538324 TI - Burning issues. PMID- 15538325 TI - Watchdogs call academies to account over conflicts of interest. PMID- 15538326 TI - Leukaemia sleuths accuse state of nuclear cover-up. PMID- 15538327 TI - Trust gives warm welcome to open access. PMID- 15538328 TI - Bush set to keep core science team for second term. PMID- 15538329 TI - Joys match fears as California agrees to stem-cell proposal. PMID- 15538330 TI - Hopes rise for RNA therapy as mouse study hits target. PMID- 15538331 TI - Herbicide critic dropped from pollution conference. PMID- 15538332 TI - WHO seeks system for tracking global clinical trials. PMID- 15538333 TI - Creative Commons ponders share options. PMID- 15538335 TI - Zoology: a mole in hand... PMID- 15538336 TI - Land remediation: Borneo is burning. PMID- 15538338 TI - Mind the gap: women racers are falling behind. PMID- 15538339 TI - Biology students find holes in gap study. PMID- 15538340 TI - Sprint research runs into a credibility gap. PMID- 15538341 TI - Linear models can't keep up with sport gender gap. PMID- 15538346 TI - Making waves. PMID- 15538347 TI - Medicine: a cholesterol connection in RNAi. PMID- 15538348 TI - Earth science: mantle mapped in the desert. PMID- 15538349 TI - DNA repair: big engine finds small breaks. PMID- 15538350 TI - Ecology: Hope in the hills for tundra? PMID- 15538351 TI - Animal behaviour: rank crime and punishment. PMID- 15538353 TI - Plant development: a bunch of leaves. PMID- 15538354 TI - Condensed-matter physics: a magnetic pendulum. PMID- 15538356 TI - Plant genetics: gene transfer from parasitic to host plants. AB - Plant mitochondrial genes are transmitted horizontally across mating barriers with surprising frequency, but the mechanism of transfer is unclear. Here we describe two new cases of horizontal gene transfer, from parasitic flowering plants to their host flowering plants, and present phylogenetic and biogeographic evidence that this occurred as a result of direct physical contact between the two. Our findings complement the discovery that genes can be transferred in the opposite direction, from host to parasite plant. PMID- 15538357 TI - Laser beams: knotted threads of darkness. AB - Destructive interference may lead to complete cancellation when light waves travelling in different directions cross, and in three-dimensional space this occurs along lines that are vortices of electromagnetic energy flow. Here we confirm theoretical predictions by experimentally creating combinations of optical laser beams in which these dark threads form stable loops that are linked and knotted. PMID- 15538358 TI - Mantle segmentation along the Oman ophiolite fossil mid-ocean ridge. AB - It has been difficult to relate the segmentation of mid-ocean ridges to processes occurring in the Earth's underlying mantle, as the mantle is rarely sampled directly and chemical variations observed in lavas at the surface are heavily influenced by details of their production as melt extracted from the mantle. Our understanding of such mantle processes has therefore relied on the analysis of pieces of fossil oceanic lithosphere now exposed at the Earth's surface, known as ophiolites. Here we present the phase chemistry and whole-rock major- and trace element contents of 174 samples of the mantle collected along over 400 km of the Oman Sultanate ophiolite. We show that, when analysed along the fossil ridge, variations of elemental ratios sensitive to the melting process define a three dimensional geometry of mantle upwellings, which can be related to the segmentation observed in modern mid-ocean ridge environments. PMID- 15538359 TI - Therapeutic silencing of an endogenous gene by systemic administration of modified siRNAs. AB - RNA interference (RNAi) holds considerable promise as a therapeutic approach to silence disease-causing genes, particularly those that encode so-called 'non druggable' targets that are not amenable to conventional therapeutics such as small molecules, proteins, or monoclonal antibodies. The main obstacle to achieving in vivo gene silencing by RNAi technologies is delivery. Here we show that chemically modified short interfering RNAs (siRNAs) can silence an endogenous gene encoding apolipoprotein B (apoB) after intravenous injection in mice. Administration of chemically modified siRNAs resulted in silencing of the apoB messenger RNA in liver and jejunum, decreased plasma levels of apoB protein, and reduced total cholesterol. We also show that these siRNAs can silence human apoB in a transgenic mouse model. In our in vivo study, the mechanism of action for the siRNAs was proven to occur through RNAi-mediated mRNA degradation, and we determined that cleavage of the apoB mRNA occurred specifically at the predicted site. These findings demonstrate the therapeutic potential of siRNAs for the treatment of disease. PMID- 15538360 TI - Crystal structure of RecBCD enzyme reveals a machine for processing DNA breaks. AB - RecBCD is a multi-functional enzyme complex that processes DNA ends resulting from a double-strand break. RecBCD is a bipolar helicase that splits the duplex into its component strands and digests them until encountering a recombinational hotspot (Chi site). The nuclease activity is then attenuated and RecBCD loads RecA onto the 3' tail of the DNA. Here we present the crystal structure of RecBCD bound to a DNA substrate. In this initiation complex, the DNA duplex has been split across the RecC subunit to create a fork with the separated strands each heading towards different helicase motor subunits. The strands pass along tunnels within the complex, both emerging adjacent to the nuclease domain of RecB. Passage of the 3' tail through one of these tunnels provides a mechanism for the recognition of a Chi sequence by RecC within the context of double-stranded DNA. Gating of this tunnel suggests how nuclease activity might be regulated. PMID- 15538361 TI - A characteristic size of approximately 10 Mpc for the ionized bubbles at the end of cosmic reionization. AB - The first galaxies to appear in the Universe at redshifts z > 20 created ionized bubbles in the intergalactic medium of neutral hydrogen left over from the Big Bang. The ionized bubbles grew with time, surrounding clusters of dwarf galaxies and eventually overlapped quickly throughout the Universe over a narrow redshift interval near z approximately 6. This event signalled the end of the reionization epoch when the Universe was a billion years old. Measuring the size distribution of the bubbles at their final overlap phase is a focus of forthcoming programmes to observe highly redshifted radio emission from atomic hydrogen. Here we show that the combined constraints of cosmic variance and light travel time imply an observed bubble size at the end of the overlap epoch of approximately 10 physical Mpc, and a scatter in the observed redshift of overlap along different lines-of sight of approximately 0.15. This scatter is consistent with observational constraints from recent spectroscopic data on the farthest known quasars. This implies that future radio experiments should be tuned to a characteristic angular scale of 0.5 degrees and have a minimum frequency bandwidth of approximately 8 MHz for an optimal detection of 21-cm flux fluctuations near the end of reionization. PMID- 15538362 TI - Strong coupling in a single quantum dot-semiconductor microcavity system. AB - Cavity quantum electrodynamics, a central research field in optics and solid state physics, addresses properties of atom-like emitters in cavities and can be divided into a weak and a strong coupling regime. For weak coupling, the spontaneous emission can be enhanced or reduced compared with its vacuum level by tuning discrete cavity modes in and out of resonance with the emitter. However, the most striking change of emission properties occurs when the conditions for strong coupling are fulfilled. In this case there is a change from the usual irreversible spontaneous emission to a reversible exchange of energy between the emitter and the cavity mode. This coherent coupling may provide a basis for future applications in quantum information processing or schemes for coherent control. Until now, strong coupling of individual two-level systems has been observed only for atoms in large cavities. Here we report the observation of strong coupling of a single two-level solid-state system with a photon, as realized by a single quantum dot in a semiconductor microcavity. The strong coupling is manifest in photoluminescence data that display anti-crossings between the quantum dot exciton and cavity-mode dispersion relations, characterized by a vacuum Rabi splitting of about 140 microeV. PMID- 15538363 TI - Vacuum Rabi splitting with a single quantum dot in a photonic crystal nanocavity. AB - Cavity quantum electrodynamics (QED) systems allow the study of a variety of fundamental quantum-optics phenomena, such as entanglement, quantum decoherence and the quantum-classical boundary. Such systems also provide test beds for quantum information science. Nearly all strongly coupled cavity QED experiments have used a single atom in a high-quality-factor (high-Q) cavity. Here we report the experimental realization of a strongly coupled system in the solid state: a single quantum dot embedded in the spacer of a nanocavity, showing vacuum-field Rabi splitting exceeding the decoherence linewidths of both the nanocavity and the quantum dot. This requires a small-volume cavity and an atomic-like two-level system. The photonic crystal slab nanocavity--which traps photons when a defect is introduced inside the two-dimensional photonic bandgap by leaving out one or more holes--has both high Q and small modal volume V, as required for strong light-matter interactions. The quantum dot has two discrete energy levels with a transition dipole moment much larger than that of an atom, and it is fixed in the nanocavity during growth. PMID- 15538364 TI - Current-induced resonance and mass determination of a single magnetic domain wall. AB - A magnetic domain wall (DW) is a spatially localized change of magnetization configuration in a magnet. This topological object has been predicted to behave at low energy as a composite particle with finite mass. This particle will couple directly with electric currents as well as magnetic fields, and its manipulation using electric currents is of particular interest with regard to the development of high-density magnetic memories. The DW mass sets the ultimate operation speed of these devices, but has yet to be determined experimentally. Here we report the direct observation of the dynamics of a single DW in a ferromagnetic nanowire, which demonstrates that such a topological particle has a very small but finite mass of 6.6 x 10(-23) kg. This measurement was realized by preparing a tunable DW potential in the nanowire, and detecting the resonance motion of the DW induced by an oscillating current. The resonance also allows low-current operation, which is crucial in device applications; a DW displacement of 10 microm was induced by a current density of 10(10) A m(-2). PMID- 15538365 TI - A fast low-power optical memory based on coupled micro-ring lasers. AB - The increasing speed of fibre-optic-based telecommunications has focused attention on high-speed optical processing of digital information. Complex optical processing requires a high-density, high-speed, low-power optical memory that can be integrated with planar semiconductor technology for buffering of decisions and telecommunication data. Recently, ring lasers with extremely small size and low operating power have been made, and we demonstrate here a memory element constructed by interconnecting these microscopic lasers. Our device occupies an area of 18 x 40 microm2 on an InP/InGaAsP photonic integrated circuit, and switches within 20 ps with 5.5 fJ optical switching energy. Simulations show that the element has the potential for much smaller dimensions and switching times. Large numbers of such memory elements can be densely integrated and interconnected on a photonic integrated circuit: fast digital optical information processing systems employing large-scale integration should now be viable. PMID- 15538366 TI - Prolonged KREEP magmatism on the Moon indicated by the youngest dated lunar igneous rock. AB - Primordial solidification of the Moon (or its uppermost layer) resulted in the formation of a variety of rock types that subsequently melted and mixed to produce the compositional diversity observed in the lunar sample suite. The initial rocks to crystallize from this Moon-wide molten layer (the magma ocean) contained olivine and pyroxene and were compositionally less evolved than the plagioclase-rich rocks that followed. The last stage of crystallization, representing the last few per cent of the magma ocean, produced materials that are strongly enriched in incompatible elements including potassium (K), the rare earth elements (REE) and phosphorus (P)--termed KREEP. The decay of radioactive elements in KREEP, such as uranium and thorium, is generally thought to provide the thermal energy necessary for more recent lunar magmatism. The ages of KREEP rich samples are, however, confined to the earliest periods of lunar magmatism between 3.8 and 4.6 billion years (Gyr) ago, providing no physical evidence that KREEP is directly involved in more recent lunar magmatism. But here we present evidence that KREEP magmatism extended for an additional 1 Gyr, based on analyses of the youngest dated lunar sample. PMID- 15538367 TI - Nonindependence of mammalian dental characters. AB - Studies of mammalian evolution frequently use data derived from the dentition. Dental characters are particularly central for inferring phylogenetic relationships of fossil taxa, of which teeth are often the only recovered part. The use of different aspects of dental morphology as phylogenetic signals implies the independence of dental characters from each other. Here we report, however, that, at least developmentally, most dental characters may be nonindependent. We investigated how three different levels of the cell signalling protein ectodysplasin (Eda) changed dental characters in mouse. We found that with increasing expression levels of this one gene, the number of cusps increases, cusp shapes and positions change, longitudinal crests form, and number of teeth increases. The consistent modification of characters related to lateral placement of cusps can be traced to a small difference in the formation of an early signalling centre at the onset of tooth crown formation. Our results suggest that most aspects of tooth shape have the developmental potential for correlated changes during evolution which may, if not taken into account, obscure phylogenetic history. PMID- 15538368 TI - Disposable-soma senescence mediated by sexual selection in an ungulate. AB - Senescence may result from an optimal balance between current reproductive investment and bodily repair processes required for future reproduction, a theoretical prediction difficult to prove especially in large, long-lived animals. Here we propose that teeth that have fixed dimensions early in life, but that wear during chewing, can be taken as a measure of total lifetime 'repair', and their wear rate as a measure of current expenditure in performance. Our approach also considers the sexual selection process to investigate the advance of senescence in males compared with females, when selection favouring competition over mates reduces the reproductive lifespan of males. We studied carcasses of 2,141 male and 739 female red deer (Cervus elaphus) of different ages, finding that male molariform teeth emerged at a far smaller size than expected from body size dimorphism. This led to higher workload, steeper wear rate and earlier depletion of male teeth than in females, in concordance with sex specific patterns of lifetime performance and reproduction. These findings provide the empirical support for the disposable-soma hypothesis of senescence, which predicts that investment in bodily repair will decrease when the return from this investment may not be realized as a result of other causes that limit survival or reproduction. PMID- 15538369 TI - A socially enforced signal of quality in a paper wasp. AB - Organisms use signals of quality to communicate information about aspects of their relative phenotypic and genetic constitution. Badges of status are a subset of signals of quality that reveal information about an individual's size and dominance. In general, signals of quality require high and differential costs to remain honest (that is, prevent low-quality cheaters from exploiting any fitness benefits associated with communicating high quality). The theoretically required costs for badges of status remain controversial because the development (or 'production') of such signals often seems to be relatively cost-free. One important hypothesis is that such signals impose social (or 'maintenance') costs incurred through repeated agonistic interactions with other individuals. However, convincing empirical evidence for social costs remains elusive. Here we report social costs in a previously undescribed badge of status: the highly variable black facial patterns of female paper wasps, Polistes dominulus. Facial patterns strongly predict body size and social dominance. Moreover, in staged contests between pairs of unfamiliar wasps, subordinate wasps with experimentally altered facial features ('cheaters') received considerably more aggression from the dominant than did sham controls, indicating that facial patterns are signals and that dishonest signalling imposes social costs. PMID- 15538370 TI - Environmental biosafety and transgenic potato in a centre of diversity for this crop. AB - The Nuffield Council on Bioethics suggests that introgression of genetic material into related species in centres of crop biodiversity is an insufficient justification to bar the use of genetically modified crops in the developing world. They consider that a precautionary approach to forgo the possible benefits invokes the fallacy of thinking that doing nothing is itself without risk to the poor. Here we report findings relevant to this and other aspects of environmental biosafety for genetically modified potato in its main centre of biodiversity, the central Andes. We studied genetically modified potato clones that provide resistance to nematodes, principal pests of Andean potato crops. We show that there is no harm to many non-target organisms, but gene flow occurs to wild relatives growing near potato crops. If stable introgression were to result, the fitness of these wild species could be altered. We therefore transformed the male sterile cultivar Revolucion to provide a genetically modified nematode-resistant potato to evaluate the benefits that this provides until the possibility of stable introgression to wild relatives is determined. Thus, scientific progress is possible without compromise to the precautionary principle. PMID- 15538371 TI - A pancreatic islet-specific microRNA regulates insulin secretion. AB - MicroRNAs (miRNAs) constitute a growing class of non-coding RNAs that are thought to regulate gene expression by translational repression. Several miRNAs in animals exhibit tissue-specific or developmental-stage-specific expression, indicating that they could play important roles in many biological processes. To study the role of miRNAs in pancreatic endocrine cells we cloned and identified a novel, evolutionarily conserved and islet-specific miRNA (miR-375). Here we show that overexpression of miR-375 suppressed glucose-induced insulin secretion, and conversely, inhibition of endogenous miR-375 function enhanced insulin secretion. The mechanism by which secretion is modified by miR-375 is independent of changes in glucose metabolism or intracellular Ca2+-signalling but correlated with a direct effect on insulin exocytosis. Myotrophin (Mtpn) was predicted to be and validated as a target of miR-375. Inhibition of Mtpn by small interfering (si)RNA mimicked the effects of miR-375 on glucose-stimulated insulin secretion and exocytosis. Thus, miR-375 is a regulator of insulin secretion and may thereby constitute a novel pharmacological target for the treatment of diabetes. PMID- 15538372 TI - In the market-place. PMID- 15538373 TI - Cell signalling: making connections. PMID- 15538374 TI - Quantum dots show their true colours. PMID- 15538375 TI - Probing real-time protein interactions. PMID- 15538377 TI - The secrets of success. PMID- 15538378 TI - Getting on the fast-track to full-time employment. PMID- 15538379 TI - Exposure assessment: utility and application within homeland or public security. PMID- 15538380 TI - The E2F family: specific functions and overlapping interests. AB - The E2F transcription factors are key regulators of cell cycle progression and the E2F field has made rapid advances since its advent in 1986. Yet, while our understanding of the roles and functions of the E2F family has made enormous progress, with each discovery new questions arise. In this review, we summarise the most recent advances in the field and discuss the remaining key questions. In particular, we will focus on how specificity is achieved among the E2Fs. PMID- 15538381 TI - The ARE-dependent mRNA-destabilizing activity of BRF1 is regulated by protein kinase B. AB - Butyrate response factor (BRF1) belongs to the Tis11 family of CCCH zinc-finger proteins, which bind to mRNAs containing an AU-rich element (ARE) in their 3' untranslated region and promote their deadenylation and rapid degradation. Independent signal transduction pathways have been reported to stabilize ARE containing transcripts by a process thought to involve phosphorylation of ARE binding proteins. Here we report that protein kinase B (PKB/Akt) stabilizes ARE transcripts by phosphorylating BRF1 at serine 92 (S92). Recombinant BRF1 promoted in vitro decay of ARE-containing mRNA (ARE-mRNA), yet phosphorylation by PKB impaired this activity. S92 phosphorylation of BRF1 did not impair ARE binding, but induced complex formation with the scaffold protein 14-3-3. In vivo and in vitro data support a model where PKB causes ARE-mRNA stabilization by inactivating BRF1 through binding to 14-3-3. PMID- 15538382 TI - FOXO transcription factor activation by oxidative stress mediated by the small GTPase Ral and JNK. AB - Forkhead transcription factors of the FOXO class are negatively regulated by PKB/c-Akt in response to insulin/IGF signalling, and are involved in regulating cell cycle progression and cell death. Here we show that, in contrast to insulin signalling, low levels of oxidative stress generated by treatment with H2O2 induce the activation of FOXO4. Upon treatment of cells with H2O2, the small GTPase Ral is activated and this results in a JNK-dependent phosphorylation of FOXO4 on threonine 447 and threonine 451. This Ral-mediated, JNK-dependent phosphorylation is involved in the nuclear translocation and transcriptional activation of FOXO4 after H2O2 treatment. In addition, we show that this signalling pathway is also employed by tumor necrosis factor alpha to activate FOXO4 transcriptional activity. FOXO members have been implicated in cellular protection against oxidative stress via the transcriptional regulation of manganese superoxide dismutase and catalase gene expression. The results reported here, therefore, outline a homeostasis mechanism for sustaining cellular reactive oxygen species that is controlled by signalling pathways that can convey both negative (PI-3K/PKB) and positive (Ras/Ral) inputs. PMID- 15538383 TI - Polyamines regulate their synthesis by inducing expression and blocking degradation of ODC antizyme. AB - Polyamines are essential organic cations with multiple cellular functions. Their synthesis is controlled by a feedback regulation whose main target is ornithine decarboxylase (ODC), the rate-limiting enzyme in polyamine biosynthesis. In mammals, ODC has been shown to be inhibited and targeted for ubiquitin independent degradation by ODC antizyme (AZ). The synthesis of mammalian AZ was reported to involve a polyamine-induced ribosomal frameshifting mechanism. High levels of polyamine therefore inhibit new synthesis of polyamines by inducing ODC degradation. We identified a previously unrecognized sequence in the genome of Saccharomyces cerevisiae encoding an orthologue of mammalian AZ. We show that synthesis of yeast AZ (Oaz1) involves polyamine-regulated frameshifting as well. Degradation of yeast ODC by the proteasome depends on Oaz1. Using this novel model system for polyamine regulation, we discovered another level of its control. Oaz1 itself is subject to ubiquitin-mediated proteolysis by the proteasome. Degradation of Oaz1, however, is inhibited by polyamines. We propose a model, in which polyamines inhibit their ODC-mediated biosynthesis by two mechanisms, the control of Oaz1 synthesis and inhibition of its degradation. PMID- 15538384 TI - Ligand-dependent switching of ubiquitin-proteasome pathways for estrogen receptor. AB - Recent evidence indicates that the transactivation of estrogen receptor alpha (ERalpha) requires estrogen-dependent receptor ubiquitination and degradation. Here we show that estrogen-unbound (unliganded) ERalpha is also ubiquitinated and degraded through a ubiquitin-proteasome pathway. To investigate this ubiquitin proteasome pathway, we purified the ubiquitin ligase complex for unliganded ERalpha and identified a protein complex containing the carboxyl terminus of Hsc70-interacting protein (CHIP). CHIP preferentially bound to misfolded ERalpha and ubiquitinated it to induce degradation. Ligand binding to the receptor induced the dissociation of CHIP from ERalpha. In CHIP-/- cells, the degradation of unliganded ERalpha was abrogated; however, estrogen-induced degradation was observed to the same extent as in CHIP+/+ cells. Our findings suggest that ERalpha is regulated by two independent ubiquitin-proteasome pathways, which are switched by ligand binding to ERalpha. One pathway is necessary for the transactivation of the receptor and the other is involved in the quality control of the receptor. PMID- 15538385 TI - Inhibition of oncogenic transformation by mammalian Lin-9, a pRB-associated protein. AB - Genetic studies in Caenorhabditis elegans identified lin-9 to function together with the retinoblastoma homologue lin-35 in vulva differentiation. We have now identified a human homologue of Lin-9 (hLin-9) and provide evidence about its function in the mammalian pRB pathway. hLin-9 binds to pRB and cooperates with pRB in flat cell formation in Saos-2 cells. In addition, hLin-9 synergized with pRB and Cbfal to transactivate an osteoblast-specific reporter gene. In contrast, hLin-9 was not involved in pRB-mediated inhibition of cell cycle progression or repression of E2F-dependent transactivation. Consistent with these data, hLin-9 was able to associate with partially penetrant pRB mutants that do not bind to E2F, but retain the ability to activate transcription and to promote differentiation. hLin-9 can also inhibit oncogenic transformation, dependent on the presence of a functional pRB protein. RNAi-mediated knockdown of Lin-9 can substitute for the loss of pRB in transformation of human primary fibroblasts. These data suggest that hLin-9 has tumor-suppressing activities and that the ability of hLin-9 to inhibit transformation is mediated through its association with pRB. PMID- 15538386 TI - Scaffolding by ERK3 regulates MK5 in development. AB - Extracellular-regulated kinase 3 (ERK3, MAPK6) is an atypical member of the ERKs, lacking the threonine and tyrosine residues in the activation loop, carrying a unique C-terminal extension and being mainly regulated by its own protein stability and/or by autophosphorylation. Here we show that ERK3 specifically interacts with the MAPK-activated protein kinase 5 (MK5 or PRAK) in vitro and in vivo. Expression of ERK3 in mammalian cells leads to nuclear-cytoplasmic translocation and activation of MK5 and to phosphorylation of both ERK3 and MK5. Remarkably, activation of MK5 is independent of ERK3 enzymatic activity, but depends on its own catalytic activity as well as on a region in the C-terminal extension of ERK3. In mouse embryonic development, mRNA expression patterns of ERK3 and MK5 suggest spatiotemporal coexpression of both kinases. Deletion of MK5 leads to strong reduction of ERK3 protein levels and embryonic lethality at about stage E11, where ERK3 expression in wild-type mice is maximum, indicating a role of this signalling module in development. PMID- 15538387 TI - Peptidoglycan recognition protein (PGRP)-LE and PGRP-LC act synergistically in Drosophila immunity. AB - In innate immunity, pattern recognition molecules recognize cell wall components of microorganisms and activate subsequent immune responses, such as the induction of antimicrobial peptides and melanization in Drosophila. The diaminopimelic acid (DAP)-type peptidoglycan potently activates imd-dependent induction of antibacterial peptides. Peptidoglycan recognition protein (PGRP) family members act as pattern recognition molecules. PGRP-LC loss-of-function mutations affect the imd-dependent induction of antibacterial peptides and resistance to Gram negative bacteria, whereas PGRP-LE binds to the DAP-type peptidoglycan, and a gain-of-function mutation induces constitutive activation of both the imd pathway and melanization. Here, we generated PGRP-LE null mutants and report that PGRP-LE functions synergistically with PGRP-LC in producing resistance to Escherichia coli and Bacillus megaterium infections, which have the DAP-type peptidoglycan. Consistent with this, PGRP-LE acts both upstream and in parallel with PGRP-LC in the imd pathway, and is required for infection-dependent activation of melanization in Drosophila. A role for PGRP-LE in the epithelial induction of antimicrobial peptides is also suggested. PMID- 15538388 TI - Interaction between human MCM7 and Rad17 proteins is required for replication checkpoint signaling. AB - Human Rad17 (hRad17) is centrally involved in the activation of cell-cycle checkpoints by genotoxic agents or replication stress. Here we identify hMCM7, a core component of the DNA replication apparatus, as a novel hRad17-interacting protein. In HeLa cells, depletion of either hRad17 or hMCM7 with small interfering RNA suppressed ultraviolet (UV) light- or aphidicolin-induced hChk1 phosphorylation, and abolished UV-induced S-phase checkpoint activation. Similar results were obtained after transfection of these cells with a fusion protein containing the hMCM7-binding region of hRad17. The hMCM7-depleted cells were also defective for the formation of ATR-containing nuclear foci after UV irradiation, suggesting that hMCM7 is required for stable recruitment of ATR to damaged DNA. These results demonstrate that hMCM7 plays a direct role in the transmission of DNA damage signals from active replication forks to the S-phase checkpoint machinery in human cells. PMID- 15538389 TI - Activation of LIMK1 by binding to the BMP receptor, BMPRII, regulates BMP dependent dendritogenesis. AB - The growth and morphological differentiation of dendrites are critical events in the establishment of proper neuronal connectivity and neural function. One extrinsic factor, BMP7, has been shown to specifically affect dendritic morphogenesis; however, the underlying mechanism by which this occurs is unknown. Here we show that LIM kinase 1 (LIMK1), a key downstream effector of Rho GTPases, colocalizes with the BMP receptor, BMPRII, in the tips of neurites and binds to BMPRII. This interaction is required for BMP-dependent induction of the dendritic arbor in cortical neurons. Furthermore, we demonstrate that the physical interaction of LIMK1 with BMPRII synergizes with the Rho GTPase, Cdc42, to activate LIMK1 catalytic activity. These studies thus define a Smad-independent pathway that directly links the BMP receptor to regulation of actin dynamics and provides insights into how extracellular signals modulate LIMK1 activity to permit fine spatial control over cytoskeletal remodelling during dendritogenesis. PMID- 15538390 TI - Longitudinal analysis of B cell repertoire and antibody gene rearrangements during early HIV infection. AB - In chronically HIV infected individuals, a number of functional B cell abnormalities have been described. However, the immediate changes that occur in the B cell compartment following viral exposure and how they affect the long-term course of infection are not well understood. We report the longitudinal analysis of B cell repertoires during early infection in untreated and treated individuals receiving highly active antiretroviral therapy (HAART). Analysis was based on IgG heavy chain gene utilization and CDR3 length measurement and relationship with CD4/CD8 counts, viral load, and total serum IgG, and anti-HIV antibodies levels. Repertoires were assessed at baseline and at weeks 2, 4, 12, 24, and 72 after initiation of therapy. The findings indicate a stable peripheral B cell repertoire during the first 72 weeks following infection, particularly in the HAART treated patients. A modest association between B cell repertoire integrity and viremia levels as well as treatment was detected. PMID- 15538391 TI - Fine mapping chromosome 16q12 in a collection of 231 systemic lupus erythematosus sibpair and multiplex families. AB - Systemic lupus erythematosus (SLE) is a chronic, autoimmune disorder influenced by multiple genetic and environmental factors. Linkage of SLE to chromosome 16q12 13 (LOD score=3.85) was first identified in pedigrees collected at the University of Minnesota, and has been replicated in several independent SLE collections. We performed fine mapping using microsatellites to further refine the susceptibility region(s), and the best evidence for linkage was identified at marker D16S3396 (LOD=2.28, P=0.0006). Evidence of association was suggested in the analysis of all families (D16S3094, P=0.0516) and improved to the level of significance (P=0.0106) when only the Caucasian families were analyzed. Subsets of pedigrees were then selected on the basis of clinical manifestations, and these subsets showed evidence for association with several markers: GATA143D05 (renal, P=0.0064), D16S3035 (renal, P=0.0418), D16S3117 (renal, P=0.0366), D16S3071 (malar rash, P=0.03638; neuropsychiatric, P=0.0349; oral ulcers, P=0.0459), D16S3094 (hematologic, P=0.0226), and D16S3089 (arthritis, P=0.0141). Together, these data provide further evidence that an important susceptibility gene(s) for SLE is located at 16q12. PMID- 15538392 TI - In vivo expression of myosin essential light chain using plasmid expression vectors in regenerating frog skeletal muscle. AB - It is well established that mutations in specific structural elements of the motor protein myosin are directly linked to debilitating diseases involving malfunctioning striated muscle cells. A potential way to study the relationship between myosin structure and function is to express exogenous myosin in vivo and determine contractile properties of the transgenic muscle cells. However, in vivo expression of functional levels of contractile proteins using transient transgenesis in skeletal muscle has not been demonstrated. Presently, we used in vivo gene transfer to express high levels of full-length myosin light chain (MLC) in skeletal muscle fibers of Rana pipiens. Anterior tibialis (AT) muscles were injected with cardiotoxin to cause degeneration and then injected at various stages of regeneration with plasmid expression vectors encoding full-length MLC1(f). In fibers from the most robustly transfected muscles 3 weeks after plasmid injections, trans-MLC1(f) expression averaged 22-43% of the endogenous MLC1(f). Trans-MLC1(f) expression was the same whether a small epitope tag was placed on the C- or N-terminus and was highly variable along individual fibers. Confocal microscopy of skinned fibers showed correct sarcomeric incorporation of trans-MLC1(f). The expression profile of myosin heavy chain isoforms 21 days after transfection was similar to normal AT muscle. These data demonstrate the feasibility of using in vivo gene transfer to probe the structural basis of contractile protein function in skeletal muscle. Based on these promising results, we discuss how further improvements in the level and consistency of myosin transgene expression may be achieved in future studies, and the therapeutic potential of plasmid gene transfer in regenerating muscle. PMID- 15538393 TI - Traumatic intracorporal septal avulsion resulting in cavernous veno-occlusive dysfunction. PMID- 15538394 TI - A prospective analysis of sexual functions during pregnancy. AB - The aim of the study was to evaluate the sexual functions during pregnancy using the Female Sexual Function Index (FSFI) questionnaire. Pregnancies were recorded in a prospective cohort study comprising 40 healthy pregnant women. Pregnant women who had a stable relationship with their partner were enrolled in the study when were first diagnosed to be pregnant. During their antenatal visits, subjects were asked to complete the FSFI questionnaire and other information about their sexual life in each trimester. Each FSFI domain score was calculated and mean scores in each domain were compared according to the trimesters of pregnancy. Data of 37 subjects for the first, 36 for the second and 34 for the third trimesters of pregnancy were eligible for the analysis. The mean age was 25.5+/ 4.5 y; mean parity was 0.4+/-0.7 and mean gravity was 1.6+/-0.9. The frequency of intercourse attempts during the last 4 weeks was 8.6+/-3 before pregnancy, and 6.9+/-2.5, 5.4+/-2.6 and 2.5+/-1.4 in the first, second and third trimesters of pregnancy, respectively. In all domains of FSFI, significant decline in domain scores was determined during pregnancy. The comparison of satisfaction and pain domain scores between first and second trimesters showed significant differences. All of the domain scores significantly decreased in the third trimester of pregnancy. Our results showed that sexual functions are significantly decreased during pregnancy and worsen as the pregnancy progresses. Childbearing couples should be given information about the sexual problems and fluctuations in the patterns of sexuality during pregnancy. PMID- 15538395 TI - Retention and migration of alprostadil cream applied topically to the glans meatus for erectile dysfunction. AB - Retention and migration of a specially designed alprostadil transdermal cream was assessed after single-dose administration to the glans meatus of the penis. Seven men were enrolled in this two-way crossover study. Three subjects self administered the radio-labeled alprostadil transdermal cream (approximately 100 mg of cream containing 300 microg alprostadil) by inserting the tip of the dispenser into the meatus. In three others, the dose was administered by dispensing the cream dropwise into the metal opening without touching the penis (the preferred and directed method). Retention and migration of the cream mass was measured in the penis with a gamma scintillation camera. The alprostadil transdermal cream was retained in the fossa navicularis at or near 99% in five of six subjects regardless of the method of dosing, thus indicating that the cream formulation performed as designed. A mild, transient burning sensation in the penis was the most common adverse event. PMID- 15538396 TI - High biochemical selectivity of tadalafil, sildenafil and vardenafil for human phosphodiesterase 5A1 (PDE5) over PDE11A4 suggests the absence of PDE11A4 cross reaction in patients. AB - The physiological role of phosphodiesterase (PDE)11 is unknown and its biochemical characteristics are poorly understood. We have expressed human His tagged PDE11A4 and purified the enzyme to apparent homogeneity. PDE11A4 displays K(m) values of 0.97 microM for cGMP and 2.4 microM for cAMP, and maximal velocities were 4- to 10-fold higher for cAMP than for cGMP. Given the homology between PDE11 and PDE5, we have compared the biochemical potencies of tadalafil (Cialis, Lilly-ICOS), vardenafil (Levitra, Bayer-GSK), and sildenafil (Viagra, Pfizer Inc.) for PDE11A4 and PDE5A1. PDE5A1/PDE11A4 selectivities are 40-, 9300-, and 1000-fold for tadalafil, vardenafil, and sildenafil, respectively. This suggests that none of these three compounds is likely to crossreact with PDE11A4 in patients. PMID- 15538397 TI - Distress: associated variables of erectile dysfunction post-acute myocardial infarction. A pilot study. AB - This study aimed to identify the associated variables of erectile dysfunction (ED) in post myocardial infarction (MI) patients without previous sexual dysfunction (SD). Out of the 37 patients, 15 (40%) had ED. Patients significantly reduced the frequency of coitus (P<0.05). Out of nine patients with distress, eight presented ED, and of 28 patients without distress, seven presented ED (89 versus 25%, P=0.001). The two groups (distress versus without distress) were similar regarding confounding variables. We concluded that patients without SD prior the MI had a significant incidence of ED and distress was an associated variable of ED post-infarction. PMID- 15538398 TI - Maternal infection and risk of cerebral palsy in term and preterm infants. AB - OBJECTIVE: We tested the hypothesis that term and preterm infants exposed to maternal infection at the time of delivery are at increased risk of developing cerebral palsy (CP). STUDY DESIGN: A population-based case-control study was conducted using Washington State birth certificate data linked to hospital discharge data. Cases (688) were children or =28 days, but not at 36 weeks PMA; moderate BPD: need for supplemental oxygen > or =28 days and <30% at 36 weeks PMA and severe BPD: need for supplemental oxygen > or =28 days, and >30% at 36 weeks PMA and/or positive pressure at 36 weeks PMA. Further, BPD-associated comorbidities and short-term outcome data during hospitalization were compared among the groups, defined by severity of BPD. RESULTS: Among LBW infants <1251 g, the incidences of BPD at 28 days and 36 weeks PMA were 21.1 and 7.4% respectively. Using the newly defined criteria, the incidences of mild, moderate and severe BPD were 13.5, 4.8 and 2.6%, respectively. In total, 64.6% of these infants had mild BPD and 70.8% weighed <751 g at birth. Associated comorbidities correlated significantly with grades of underlying pulmonary disease. Also, significantly longer hospital stay, discharge at a higher PMA and lower growth velocity was observed with increasing grades of BPD. CONCLUSIONS: The new system for grading the severity of BPD offers a better description of underlying pulmonary disease and correlates with the infant's maturity, growth and overall severity of illness. Whether it will have a role in predicting long-term outcome remains to be determined. PMID- 15538400 TI - Mutations in the BRAF and N-ras genes in childhood acute lymphoblastic leukaemia. PMID- 15538401 TI - Ploidy, as detected by fluorescence in situ hybridization, defines different subgroups in multiple myeloma. AB - Ploidy appears as an important parameter in both the biology and the clinical evolution of multiple myeloma. However, its evaluation requires either a successful karyotyping (obtained in 30% of the patients) or a DNA index calculation by flow cytometry (not routinely performed in myeloma). We validated a novel method based on interphase fluorescence in situ hybridization that can be utilitized to analyze almost all the patients. The method was very specific and sensitive for the detection of hyperdiploidy. Extended studies showed that most recurrent 14q32 translocations occur in nonhyperdiploid clones, and that deletions of chromosome 13 were less frequently observed in hyperdiploid clones (48 vs 66%). Further large studies are ongoing to evaluate the prognostic value of ploidy in myeloma. PMID- 15538402 TI - Arsenic trioxide (ATO) and MEK1 inhibition synergize to induce apoptosis in acute promyelocytic leukemia cells. AB - Recent studies suggest that components of the prosurvival signal transduction pathways involving the Ras-mitogen-activated protein kinase (MAPK) can confer an aggressive, apoptosis-resistant phenotype to leukemia cells. In this study, we report that acute promyelocytic leukemia (APL) cells exploit the Ras-MAPK activation pathway to phosphorylate at Ser112 and to inactivate the proapoptotic protein Bad, delaying arsenic trioxide (ATO)-induced apoptosis. Both in APL cell line NB4 and in APL primary blasts, the inhibition of extracellular signal regulated kinases 1/2 (ERK1/2) and Bad phosphorylation by MEK1 inhibitors enhanced apoptosis in ATO-treated cells. We isolated an arsenic-resistant NB4 subline (NB4-As(R)), which showed stronger ERK1/2 activity (2.7-fold increase) and Bad phosphorylation (2.4-fold increase) compared to parental NB4 cells in response to ATO treatment. Upon ATO exposure, both NB4 and NB4-As(R) cell lines doubled protein levels of the death antagonist Bcl-xL, but the amount of free Bcl xL that did not heterodimerize with Bad was 1.8-fold greater in NB4-As(R) than in the parental line. MEK1 inhibitors dephosphorylated Bad and inhibited the ATO induced increase of Bcl-xL, overcoming ATO resistance in NB4-As(R). These results may provide a rationale to develop combined or sequential MEK1 inhibitors plus ATO therapy in this clinical setting. PMID- 15538403 TI - Biclonal blast crisis with a mutated ABL catalytic domain in a Ph, del (9q) positive CML patient responsive to imatinib: drug resistance should be monitored in all patients irrespective of response status. PMID- 15538404 TI - Hyperleukocytosis complicating lonafarnib treatment in patients with chronic myelomonocytic leukemia. PMID- 15538405 TI - Drug-induced immunophenotypic modulation in childhood ALL: implications for minimal residual disease detection. AB - Assessment of minimal residual disease (MRD) by flow cytometry is considered to be based on the reproducibility of the leukemic immunophenotype detected at diagnosis. However, we previously noticed modulation of surface antigen expression in acute lymphoblastic leukemia (ALL) during the early treatment. Hence, we investigated this in 30 children with B-cell precursor ALL consecutively enrolled in the AIEOP-BFM ALL 2000 protocol. Quantitative expression of seven antigens useful in MRD monitoring was studied at diagnosis and compared to that measured at different time points of remission induction therapy. Downmodulation in the expression of CD10 and CD34 occurred at follow-up. By contrast, upmodulation of CD19, CD20, CD45RA, and CD11a was observed, while the expression of CD58 remained stable. Despite this, we could unambiguously discriminate leukemic cells from normal residual B cells. This holds true when bone marrow (BM) samples from similarly treated T-ALL patients, but not from healthy donors, were used as reference. Our results indicate that immunophenotypic modulation occurs in ALL during the early phases of BFM-type protocols. However, the accuracy of MRD detection by flow cytometry seems not negatively affected if adequate analysis protocols are employed. Investigators should take this phenomenon into account in order to avoid pitfalls in flow cytometric MRD studies. PMID- 15538406 TI - Genetic heterogeneity in multiple myeloma. AB - In the past decade, many progresses have been made in our knowledge of the genetics of multiple myeloma. The use of molecular cytogenetic techniques has led to the identification of several recurrent (cyto)genetic abnormalities, representing either prognostic markers, or novel therapeutic targets. More global analyses of this genetic heterogeneity using expression array technologies should further extend our understanding of the disease, hopefully enabling some improvements in the treatment of the patients. The goal of this minireview is to summarize these recent advances. PMID- 15538408 TI - XXIX Congress of the Czech and Slovak Gastroenterological and Hepatological Societies, Olomouc, 2-4 October 2003. Abstracts. PMID- 15538407 TI - Th1-biased humoral immune responses against Wilms tumor gene WT1 product in the patients with hematopoietic malignancies. AB - The Wilms' tumor gene WT1 is highly expressed in leukemias and myelodysplastic syndrome (MDS), and WT1 expression levels increase along with the disease progression in chronic myeloid leukemia and MDS. We previously reported that IgM and IgG WT1 antibodies were detected with significantly higher detection rate and antibody titers in leukemias and MDS compared to those in healthy volunteers. In this study, whether IgG humoral immune responses against WT1 protein were Th1- or Th2-type were determined by measurement of four subclasses of IgG WT1 antibody, IgG1, IgG2, IgG3, and IgG4. In leukemias and MDS, Th1-type WT1 antibodies such as IgG1, IgG2, and IgG3 were significantly increased in both detection rate and antibody titers compared to those in healthy volunteers, whereas Th2-type WT1 antibody such as IgG4 did not increase. These results showed that Th1-biased humoral immune responses against WT1 protein were generated in leukemias and MDS. These results should allow us to consider that Th1-biased cellular immune responses against WT1 protein, which was essentially needed for cancer immunotherapy targeting WT1, should be elicited in patients with hematopoietic malignancies. PMID- 15538538 TI - Convergent evolution of chromosomal sex-determining regions in the animal and fungal kingdoms. AB - Sexual identity is governed by sex chromosomes in plants and animals, and by mating type (MAT) loci in fungi. Comparative analysis of the MAT locus from a species cluster of the human fungal pathogen Cryptococcus revealed sequential evolutionary events that fashioned this large, highly unusual region. We hypothesize that MAT evolved via four main steps, beginning with acquisition of genes into two unlinked sex-determining regions, forming independent gene clusters that then fused via chromosomal translocation. A transitional tripolar intermediate state then converted to a bipolar system via gene conversion or recombination between the linked and unlinked sex-determining regions. MAT was subsequently subjected to intra- and interallelic gene conversion and inversions that suppress recombination. These events resemble those that shaped mammalian sex chromosomes, illustrating convergent evolution in sex-determining structures in the animal and fungal kingdoms. PMID- 15538539 TI - Enzyme immobilization onto micro-sized modified PMMA microspheres. AB - Micro-sized acrolein-modified PMMA (poly[methyl methacrylate]) microspheres were used as the support for enzyme immobilization. The properties of immobilized enzyme as well as the immobilizing behavior were studied. The immobilized amount reached 76.8 mg.g(-1) support, which is relatively high compared with some analogous supports. The K(m) value was larger and the V(max) was smaller in the immobilized form than in the free form. Better resistance of the immobilized enzyme against the temperature, time and pH changes showed that the modified PMMA microspheres could be a good support for immobilization of pepsin. PMID- 15538540 TI - Behavioral profile of SB 269970, a selective 5-HT(7) serotonin receptor antagonist, in social encounters between male mice. AB - The 5-HT(7) receptor is targeted by several antipsychotics, such as pimozide or clozapine, which have demonstrated antiaggressive properties in laboratory animal models. Likewise, 'in situ' hybridization and autoradiography studies have revealed moderate to high densities of 5-HT(7) receptors in limbic areas and hypothalamus, which have long been involved in the control of aggression. However, to date there is no evidence concerning the role of this receptor in the regulation of aggression. This study was designed to examine the effects of SB 269970 (1, 2, 4, 8, 16 and 32 mg/kg, i.p.), a selective 5-HT(7) receptor antagonist, on agonistic behavior elicited by isolation in mice. Individually housed mice were exposed to anosmic standard opponents 15 min after drug administration and encounters were videotaped and evaluated using an ethopharmacologically-based analysis. The results indicated that SB 269970 did not produce any significant behavioral changes, suggesting that 5-HT(7) receptors might not be involved in the modulation of aggression. Further studies with other selective ligands for these receptors are needed to confirm these findings. PMID- 15538541 TI - The effects of the sulfonylurea glyburide on glutathione peroxidase, superoxide dismutase and catalase activities in the heart tissue of streptozotocin-induced diabetic rat. AB - Oxygen free radicals have been suggested to be a contributory factor in diabetes complications. The aim of this study was to examine the effects of glyburide on the antioxidant enzyme activities in the heart tissue of diabetic rats. We investigated the activities of antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) in the hearts of both control and streptozotocin-induced diabetic rats. In the heart of diabetic rats, the activity of total superoxide dismutase decreased significantly (p < 0.005), whereas the activity of catalase and glutathione peroxidase increased to a large extent (p < 0.0001 and p = 0.05, respectively) at the end of the fourth week compared with the control group. Glyburide treatment of diabetic rats for 4 weeks corrected the changes observed in diabetic heart. In addition, blood glucose levels of untreated diabetic rats decreased following the glyburide treatment. These results demonstrate that the sulfonylurea glyburide is capable of exerting direct insulin-like effect on heart superoxide dismutase, catalase and glutathione peroxidase activities of diabetic rats in vivo. PMID- 15538542 TI - Association between APOE epsilon4 allele and increased expression of CD95 on T cells from patients with Alzheimer's disease. AB - Alzheimer's disease (AD) is the most prevalent neurodegenerative disorder, characterized by progressive impairment of cognitive functions. AD has a strong and complex genetic etiology, and multiple genes, acting independently and/or interacting, likely to influence the risk of developing AD. To test whether the expression of Fas receptor is upregulated in peripheral blood T lymphocytes and whether or not it correlates with APOE genotypes, 88 patients with AD and 24 normal individuals as controls were included in this study. T lymphocytes from patients as opposed to controls did undergo DNA fragmentation after in vitro exposure to IgM anti-Fas. In addition, several activation markers (CD25, HLA-DR, and CD45R0) were increased after 72 h in culture with respect to the controls, and Fas expression was also significantly different from the control group (p < 0.01). Reverse transcription PCR for Fas mRNA yielded the same results. T cells from both patients and controls showed upregulation of Fas receptor expression after in vitro anti-CD3 stimulation. Co-culture experiments with interleukin-4 downmodulated surface Fas receptor expression on T cells from patients and at a lesser extent in the control group. AD patients with the APOE allele 4 showed an increased expression of CD95 (53% +/- 6) with respect to APOE allele 3 (38% +/- 4). The control group showed a 22% +/- 3 (allele 4) and 31% +/- 5 (allele 3), respectively. Hyperexpression of Fas mRNA and surface Fas receptor on CD45RO(+) T lymphocytes may explain the occurrence of inflammatory cellular infiltrates in the CNS of AD patients. PMID- 15538543 TI - Therapeutic drug monitoring of non-anticancer drugs in cancer patients. AB - Therapeutic drug monitoring (TDM) or the measurement of drug concentrations in plasma, serum or blood, aims to improve clinical activity, avoid toxicity, and reduce the costs of drug treatment. Specific conditions for TDM to be reasonably applied include the availability of a validated assay, a considerable interindividual pharmacokinetic variability, a high correlation between drug concentration and toxicity, and a narrow therapeutic index. Cancer patients are especially prone to drug-drug interactions due to significant comedication, impaired liver and kidney function and hypoalbuminemia with altered drug binding. This article discusses TDM for various broadly used non-anticancer drugs in cancer patients and gives specific recommendations. Selected drugs covered in this article include those regularly used in febrile neutropenic patients such as the glycopeptide antibiotics, aminoglycosides, the antifungal agents, including flucytosine and azole compounds, the anticonvulsants phenytoin, carbamazepine and valproate, the tricyclic antidepressants, selective-serotonin-reuptake inhibitors, lithium, morphine, digitalis glycosides and the immunosupressants cyclosporin A, tacrolimus, sirolimus and mycophenolate mofetil, crucial compounds in the setting of bone marrow transplantation. In all cases, treating physicians have to consider the variability in patient age, disease stage, comedication, organ function and protein level to weigh the pros and cons of TDM in the individual cancer patient. PMID- 15538544 TI - Pharmacology of catamenial epilepsy. AB - Catamenial epilepsy is a menstrual cycle-related seizure disorder characterized by an increase in seizures at the time of menstruation. Catamenial epilepsy affects up to 70% of women with epilepsy. Catamenial seizures are common among women with focal or generalized epilepsy, which affects an estimated 1 million women in the United States. Presently, there is no specific, FDA-approved drug treatment for catamenial epilepsy. Despite the increased use of wide-ranging antiepileptic and hormonal drugs, catamenial seizures are often refractory to many treatments. Recent studies have provided an improved understanding of the pathophysiology of catamenial epilepsy. Cyclical changes of ovarian hormones estrogens and progesterone are now widely believed to be essential for the genesis of catamenial seizures. Generally, progesterone has antiseizure effects, while estrogens facilitate seizure susceptibility. The progesterone metabolite allopregnanolone has been identified as a key endogenous neurosteroid with powerful antiseizure activity. Allopregnanolone is a potent, positive allosteric modulator of GABA(A) receptors. Progesterone and allopregnanolone exposure and withdrawal affects GABA(A) receptor plasticity. In animal models, withdrawal from chronic progesterone and, consequently, of allopregnanolone levels in brain, has been shown to increase seizure susceptibility. Natural progesterone therapy is proven to be effective in women with epilepsy. Consequently, synthetic neurosteroids that are devoid of hormonal side effects represent a novel class of antiepileptic drugs for women with catamenial epilepsy. Our studies suggest that ganaxolone, a GABA(A) receptor-modulating synthetic neuroactive steroid, is a particularly promising treatment for catamenial epilepsy. Future studies are clearly warranted to determine the molecular pathophysiology and an effective treatment of catamenial epilepsy. PMID- 15538545 TI - Acyl coenzyme A:cholesterol acyltransferase inhibitors as hypolipidemic and antiatherosclerotic drugs. AB - Acyl coenzyme A:cholesterol acyltransferase (ACAT) is the enzyme that catalyzes the conversion of intracellular cholesterol into cholesteryl esters. Two ACAT isoforms, termed ACAT1 and ACAT2, have been described. ACAT1 is ubiquitously found, with high expression levels in macrophages, adrenals, sebaceous glands and foam cells from human atherosclerotic lesions. In contrast, ACAT2 expression is restricted to the intestine and the liver of mice and non-human primates. The reaction catalyzed by ACAT is essential for intestinal cholesterol absorption, synthesis and secretion of apolipoprotein B (apoB)-containing lipoproteins, and intracellular storage of cholesterol. Therefore, ACAT inhibitors would theoretically reduce plasma cholesterol levels by blocking cholesterol absorption from the diet and by reducing hepatic VLDL synthesis. Moreover, ACAT inhibition could limit the accumulation of cholesteryl esters in the cytoplasm of macrophages, thus reducing the formation of foam cells. In view of these attractive possibilities, a great deal of molecules with ACAT inhibitory properties have been synthesized in the last 20 years. However, only a few of them have reached clinical studies, mainly due to unexpected side effects. On the other hand, most of the compounds assayed in humans have not shown substantial hypolipidemic efficacy. The present article focuses on the current knowledge of the pharmacology of ACAT inhibitors, and, specifically, on the different pharmacological approaches used to evaluate these compounds as hypolipidemic and antiatherosclerotic agents. PMID- 15538546 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables has been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: 101M, 166Ho-DOTMP, 3-AP; Abatacept, abetimus sodium, ACR-16, adefovir dipivoxil, alefacept, AMD-070, aminolevulinic acid hexyl ester, anatumomab mafenatox, anti-CTLA-4 MAb, antigastrin therapeutic vaccine, AP-12009, AP-23573, APC-8024, aripiprazole, ATL-962, atomoxetine hydrochloride; Bevacizumab, bimatoprost, bortezomib, bosentan, BR-1; Calcipotriol/betamethasone dipropionate, cinacalcet hydrochloride, clofazimine, colchicine, cold-adapted influenza vaccine trivalent, CRM197; Desloratadine, desoxyepothilone B, diethylhomospermine; Edodekin alfa, efalizumab, elcometrine, eletriptan, enfuvirtide, entecavir, EP-2101, eplerenone, erlotinib hydrochloride, etoricoxib, everolimus, exherin, ezetimibe; Febuxostat, fluorescein lisicol, fosamprenavir calcium, frovatriptan; Hemoglobin raffimer, HSPPC-96, human insulin; Imatinib mesylate, insulin detemir, insulin glargine, IRX-2, istradefylline, IV gamma-globulin, ixabepilone; Kahalalide F; L-759274, levodopa/carbidopa/entacapone, licofelone, lonafarnib, lopinavir, lurtotecan, LY 156735; MAb G250, mecasermin, melatonin, midostaurin, muraglitazar; Nesiritide, nitronaproxen; O6-Benzylguanine, olmesartan medoxomil, olmesartan medoxomil/hydrochlorothiazide, omapatrilat, oral insulin; Parecoxib sodium, PCK 3145, peginterferon alfa-2a, peginterferon alfa-2b, peginterferon alfa-2b/ ribavirin, pemetrexed disodium, peptide YY3-36, PG-CPT, phenoxodiol, pimecrolimus, posaconazole; Rasagiline mesilate, rDNA insulin, RG228, rimonabant hydrochloride, rosuvastatin calcium, rotigotine hydrochloride; S-3304, safinamide mesilate, salcaprozic acid sodium salt, SDZ-SID-791, SGN-30, soblidotin, squalamine; Telmisartan/hydrochlorothiazide, testosterone gel, TF(c)-KLH conjugate vaccine, TH-9507, theraloc, tipifarnib, tocilizumab, travoprost; ValboroPro, valdecoxib, veglin, voriconazole; Ximelagatran. PMID- 15538547 TI - The genetics of bulimia nervosa. AB - Although historically bulimia nervosa has been considered to be a disorder influenced primarily by sociocultural forces such as societal emphasis on thinness and attractiveness, over the past decade it has become increasingly clear that biology and genetics contribute substantially to its etiology. Currently, it is believed that combined study of genetic and environmental risk factors may ultimately hold the clue to the question of why when so many young individuals are exposed to societal risk factors, only a small number actually develop frank eating disorders. Genetic research on eating disorders is progressing in the direction of enhanced explication of the phenotype by focusing on behavioral, neurobiological and temperamental variables that may represent core features of the disorder. Such enhanced phenotyping may assist with identifying areas of the genome that may be implicated in susceptibility to these serious and debilitating conditions. PMID- 15538548 TI - Fulvestrant: a new type of estrogen receptor antagonist for the treatment of advanced breast cancer. AB - Postmenopausal women with hormone receptor- positive tumors are candidates for endocrine treatment. Current treatment options include the selective estrogen receptor modulators (e.g., tamoxifen and toremifene), which inhibit estrogen receptor signaling, and the aromatase inhibitors (e.g., anastrozole and letrozole), which prevent the conversion of androgens into estrogen in postmenopausal women. As most patients eventually become resistant to endocrine agents, there is a need for new treatments that are effective, well tolerated and lack cross-resistance with currently available therapies. This review describes the development of fulvestrant (Faslodex), a new type of endocrine agent, which is an estrogen receptor antagonist with no agonist effects. Phase III clinical trials have found that fulvestrant is as effective and well tolerated as anastrozole for treating postmenopausal patients with advanced breast cancer who have progressed on one prior endocrine therapy. In addition, fulvestrant has first-line efficacy similar to that of tamoxifen in patients with estrogen receptor-positive and/or progesterone receptor-positive breast cancer. Moreover, in a compassionate-use program, it has become clear that fulvestrant is not cross resistant with other therapies. Therefore, fulvestrant is a versatile new treatment option for postmenopausal women with advanced breast cancer who have progressed on prior endocrine therapy. PMID- 15538549 TI - Pharmacotherapy in lower back pain. AB - In the US, back pain is the second most common cause of disability and the leading cause among men, with approximately 45% of the adult population experiencing lower back pain annually and a direct cost for diagnosis and treatment reported to be higher than 23 billion US dollars in 1990. However, despite the high prevalence of this condition, lower back pain diagnoses are commonly imprecise, and specific causes for lower back pain can only be determined in approximately 15% of patients. So, although for most patients with acute lower back pain, a simple cause-and-effect model can be described, often the result of a lumbar sprain or strain, clinicians must be alert to a variety of other conditions which may present in a similar fashion and require more emergent care. Pharmacotherapy and nondrug-related modalities have been shown to reduce pain and other related symptoms. Medication classes with known benefit include the nonsteroidal antiinflammatory drugs (NSAIDs), skeletal muscle relaxants, opioids, acetaminophen and the newer cyclooxygenase-2 (COX-2) inhibitors. This review analyzes the different drugs available for treating lower back pain in light of the most recent evidence coming from clinical studies. More critical research is needed to further define the roles of these medications in treating pain associated with lower back injury. PMID- 15538550 TI - Duloxetine: a new serotonin/noradrenaline reuptake inhibitor for the treatment of depression. AB - Double-blind, placebo-controlled clinical trials have evaluated and demonstrated the efficacy of duloxetine as an antidepressant in patients with major depressive disorders. The drug has been noted to be well tolerated and effective in the control of depressive symptoms. In addition, duloxetine has been shown to be better than placebo and as effective as paroxetine as an antidepressant and also better than placebo for reducing pain in both experimental models and patients. Duloxetine is a safe and well-tolerated new treatment option for depression including anxiety and painful physical symptoms. Furthermore, duloxetine has proven robust efficacy in stress urinary incontinence. PMID- 15538551 TI - Stroke prevention with losartan in the context of other antihypertensive drugs. AB - Several large clinical trials have demonstrated that successful control of blood pressure decreases the incidence of strokes. Also, drugs that stimulate the production of angiotensin II, such as diuretics, calcium channel blockers (CCBs) and angiotensin receptor blockers (ARBs), provide additional stroke reduction than drugs that suppress angiotensin II production such as beta-blockers and angiotensin converting enzyme (ACE) inhibitors. Since the stroke-protective effect of angiotensin II is mediated through stimulation of the AT2 receptors, drugs that selectively block the AT1, such as the ARBs, provide greater stroke protection than the other antihypertensive drugs. The blockade of the AT1 receptors lessens local brain ischemia, whereas the stimulation of the AT2 receptors increases local blood flow through recruitment of collateral vessels. Among the ARBs, losartan possesses certain unique properties not shared by other members of its class, which enhance its stroke-protective effects. These include the prevention of platelet adhesiveness and aggregation and the decrease of serum uric acid levels, which both lead to reduction in cardiovascular and cerebrovascular events. ( PMID- 15538552 TI - Treatment of H2S using a horizontal biotrickling filter based on biological activated carbon: reactor setup and performance evaluation. AB - Biological treatment is an emerging and prevalent technology for treating off gases from wastewater treatment plants. The most commonly reported odorous compound in off-gases is hydrogen sulfide (H(2)S), which has a very low odor threshold. A self-designed, bench-scale, cross-flow horizontal biotrickling filter (HBF) operated with bacteria immobilized activated carbon (termed biological activated carbon-BAC), was applied for the treatment of H(2)S. A mixed culture of sulfide-oxidizing bacteria dominated by Acidithiobacillus thiooxidans acclimated from activated sludge was used as bacterial seed and the biofilm was developed by culturing the bacteria in the presence of carbon pellets in mineral medium. HBF performance was evaluated systematically over approximately 120 days, depending on a series of changing factors including inlet H(2)S concentration, gas retention time (GRT), pH of recirculation solution, upset and recovery, sulfate accumulation, pressure drop, gas-liquid ratio, and shock loading. The biotrickling filter system can operate at high efficiency from the first day of operation. At a volumetric loading of 900 m(3) m(-3) h(-1) (at 92 ppmv H(2)S inlet concentration), the BAC exhibited maximum elimination capacity (113 g H(2)S/m(-3) h(-1)) and a removal efficiency of 96% was observed. If the inlet concentration was kept at around 20 ppmv, high H(2)S removal (over 98%) was achieved at a GRT of 4 s, a value comparable with those currently reported for biotrickling filters. The bacterial population in the acidic biofilter demonstrated capacity for removal of H(2)S over a broad pH range (pH 1-7). A preliminary investigation into the different effects of bacterial biodegradation and carbon adsorption on system performance was also conducted. This study shows the HBF to be a feasible and economic alternative to physical and chemical treatments for the removal of H(2)S. PMID- 15538553 TI - Growth rates of Dekkera/Brettanomyces yeasts hinder their ability to compete with Saccharomyces cerevisiae in batch corn mash fermentations. AB - Growth rates determined by linear regression analysis revealed that Saccharomyces cerevisiae consistently grew more rapidly than Brettanomyces yeasts under a wide array of batch fermentative conditions, including acetic acid stress, in normal gravity (ca. 20 degrees Plato) mashes made from ground corn. Brettanomyces yeasts only grew more rapidly than S. cerevisiae when acetic acid concentrations were elevated to industrially irrelevant levels (>0.45%, w/v). Furthermore, the three Brettanomyces isolates used in this study failed to produce significant quantities of acetic acid under pure culture fermentative conditions. In fact, the small amounts of acetic acid which accumulated in pure culture fermentations of whole corn mash were below the concentration required to inhibit the growth and metabolism of S. cerevisiae. Acetic acid concentrations in pure culture Brettanomyces fermentations exceeded 0.05% (w/v) only in media containing low levels of glucose (<4%, w/v) or when aeration rates were elevated to at least 0.03 vol. air vol.-1 mash min-1. Consequently, it was concluded that Brettanomyces yeasts would not be capable of competing with S. cerevisiae in industrial batch fermentations of whole corn mash based solely on growth rates, nor would they be capable of producing inhibitory concentrations of acetic acid in such fermentations. PMID- 15538554 TI - Purification, cloning, and properties of alpha-galactosidase from Saccharopolyspora erythraea and its use as a reporter system. AB - An alpha-galactosidase from the erythromycin-producing bacterium Saccharopolyspora erythraea was purified to near homogeneity. The enzyme has an apparent molecular mass of 45 kDa as determined by SDS-PAGE. The pH optimum, K(m) for p-nitrophenyl-alpha-D: -glucopyranoside (pNPalphaG), K(m) for melibiose and the V(max) are similar to those of other studied alpha-galactosidase enzymes. The N-terminal amino-acid sequence of this protein was determined. PCR amplification was used to generate a 640-bp product using oligonucleotide primers based on the N-terminal amino-acid sequence and a downstream region that is conserved in other related alpha-galactosidase enzymes. This fragment was used as a probe to clone the alpha-galactosidase gene, designated melA, from a S. erythraea lambda phage chromosomal library. S. erythraea appears to possess an unique alpha galactosidase enzyme, encoded by melA, that can utilize galactopyranosides as carbon sources. Furthermore, the ability to use the product of melA as a reporter enzyme in S. erythraea has been demonstrated. The alpha-galactosidase uses the substrates 5-bromo-4-chloro-3-indoyl-alpha-D: -galactosidase (X-alpha-gal) on agar media and pNPalphaG in liquid media. PMID- 15538555 TI - A novel fungal omega3-desaturase with wide substrate specificity from arachidonic acid-producing Mortierella alpina 1S-4. AB - A filamentous fungus, Mortierella alpina 1S-4, is capable of producing not only arachidonic acid (AA; 20:4n-6) but also eicosapentaenoic acid (EPA; 20:5n-3) below a cultural temperature of 20 degrees C. Here, we describe the isolation and characterization of a gene (maw3) that encodes a novel omega3-desaturase from M. alpina 1S-4. Based on the conserved sequence information for M. alpina 1S-4 Delta12-desaturase and Saccharomyces kluyveri omega3-desaturase, the omega3 desaturase gene from M. alpina 1S-4 was cloned. Homology analysis of protein databases revealed that the amino acid sequence showed 51% identity, at the highest, with M. alpina 1S-4 Delta12-desaturase, whereas it exhibited 36% identity with Sac. kluyveri omega3-desaturase. The cloned cDNA was confirmed to encode the omega3-desaturase by its expression in the yeast Sac. cerevisiae. Analysis of the fatty acid composition of the yeast transformant demonstrated that 18-carbon and 20-carbon n-3 polyunsaturated fatty acids (PUFAs) were accumulated through conversion of exogenous 18-carbon and 20-carbon n-6 PUFAs. The substrate specificity of the M. alpina 1S-4 omega3-desaturase differs from those of the known fungal omega3-desaturases from Sac. kluyveri and Saprolegnia diclina. Plant, cyanobacterial and Sac. kluyveri omega3-desaturases desaturate 18 carbon n-6 PUFAs, Spr. diclina omega3-desaturase desaturates 20-carbon n-6 PUFAs and Caenorhabditis elegans omega3-desaturase prefers 18-carbon n-6 PUFAs as substrates rather than 20-carbon n-6 PUFAs. The substrate specificity of M. alpina 1S-4 omega3-desaturase is rather similar to that of C. elegans omega3 desaturase, but the M. alpina omega3-desaturase can more effectively convert AA into EPA when expressed in yeast. The M. alpina 1S-4 omega3-desaturase is the first known fungal desaturase that uses both 18-carbon and 20-carbon n-6 PUFAs as substrates. PMID- 15538556 TI - Towards electronic paper displays made from microbial cellulose. AB - Cellulose (in the form of printed paper) has always been the prime medium for displaying information in our society and is far better than the various existing display technologies. This is because of its high reflectivity, contrast, low cost and flexibility. There is a major initiative to push for a dynamic display technology that emulates paper (popularly known as "electronic paper"). We have successfully demonstrated the proof of the concept of developing a dynamic display on cellulose. To the best of our knowledge, this is the first significant effort to achieve an electronic display using bacterial cellulose. First, bacterial cellulose is synthesized in a culture of Acetobacter xylinum in standard glucose-rich medium. The bacterial cellulose membrane thus formed (not pulp) is dimensionally stable, has a paper-like appearance and has a unique microfibrillar nanostructure. The technique then involves first making the cellulose an electrically conducting (or semi-conducting) sheet by depositing ions around the microfibrils to provide conducting pathways and then immobilizing electrochromic dyes within the microstructure. The whole system is then cased between transparent electrodes, and upon application of switching potentials (2-5 V) a reversible color change can be demonstrated down to a standard pixel-sized area (ca. 100 microm2). Using a standard back-plane or in-plane drive circuit, a high-resolution dynamic display device using cellulose as substrate can be constructed. The major advantages of such a device are its high paper-like reflectivity, flexibility, contrast and biodegradability. The device has the potential to be extended to various applications, such as e-book tablets, e newspapers, dynamic wall papers, rewritable maps and learning tools. PMID- 15538557 TI - Biotechnology of desulfurization of diesel: prospects and challenges. AB - To meet stringent emission standards stipulated by regulatory agencies, the oil industry is required to make a huge investment to bring down the sulfur content in diesel to the desired level, using conventional hydrodesulfurization (HDS) technology, by which sulfur is catalytically converted to hydrogen sulfide in the presence of hydrogen. These reactions proceed rapidly only at high temperature and pressure and therefore the capital cost as well as the operating cost associated with HDS very high. Biological desulfurization has the potential of being developed as a viable technology downstream of classical HDS. Various attempts have been made to develop biotechnological processes based on microbiological desulfurization employing aerobic and anaerobic bacteria. However, there are several bottlenecks limiting commercialization of the process. This review discusses various aspects of microbial desulfurization and the progress made towards its commercialization. PMID- 15538558 TI - Molecular characterization of a beta-1,4-endoglucanase from an endophytic Bacillus pumilus strain. AB - Endophytes comprise mainly microorganisms that colonize inner plant tissues, often living with the host in a symbiotic manner. Several ecological roles have been assigned to endophytic fungi and bacteria, such as antibiosis to phytopathogenic agents and plant growth promotion. Nowadays, endophytes are viewed as a new source of genes, proteins and biochemical compounds that may be used to improve industrial processes. In this study, the gene EglA was cloned from a citrus endophytic Bacillus strain. The EglA encodes a beta-1,4 endoglucanase capable of hydrolyzing cellulose under in vitro conditions. The predicted protein, EglA, has high homology to other bacterial cellulases and shows a modular structure containing a catalytic domain of the glycosyl hydrolase family 9 (GH9) and a cellulose-binding module type 3 (CBM3). The enzyme was expressed in Escherichia coli, purified to homogeneity, and characterized. EglA has an optimum pH range of 5-8, and remarkable heat stability, retaining more than 85% activity even after a 24-h incubation at pH 6-8.6. This characteristic is an important feature for further applications of this enzyme in biotechnological processes in which temperatures of 50-60 degrees C are required over long incubation periods. PMID- 15538559 TI - Manganese peroxidase of Agaricus bisporus: grain bran-promoted production and gene characterization. AB - The main manganese peroxidase (MnP) isoenzyme of Agaricus bisporus ATCC 62459 produced in lignocellulose-containing cultures was isolated, cloned and sequenced. In liquid medium, where MnP was previously detected only in trace amounts, the production of MnP was enhanced by rye and wheat bran supplements. The pI (3.25) and N-terminal amino acid sequence (25 aa) of the enzyme from bran containing cultures were identical to those reported from compost-isolated MnP1. MnP1 is a 328-aa long polypeptide preceded by a 26-aa leader peptide. The nucleotide sequence and putative amino acid sequence of MnP1 reveal its similarity to Pleurotus ostreatus MnP3 (62.5%), Lepista irina versatile peroxidase (VP) (61.8%) and Pleurotus eryngii VPs VPL2 and VPL1 (61.9% and 61.2%, respectively). The intron-exon structure resembles that of P. ostreatus MnP1 and P. eryngii VPL1. Despite the sequence similarity to VPs, in the A. bisporus MnP1 sequence, alanine (A163) is present instead of tryptophane (W164), distinguishing it from the veratryl alcohol oxidising P. eryngii VPLs. The MnP sequence can be used as a tool to examine the pattern of ligninolytic gene expression during the growth and fruiting of A. bisporus to optimise compost composition, fungal growth and mushroom production. PMID- 15538560 TI - Magnetic resonance imaging of intramuscular myxoma with histological comparison and a review of the literature. AB - OBJECTIVE: To evaluate the magnetic resonance (MR) features of intramuscular myxoma (IM) compared with its pathological findings. DESIGN: Two radiologists retrospectively reviewed records and imaging studies of patients with histologically proven IM. Two radiologists also analyzed by consensus all the MR studies (pre- and post-contrast T1-weighted and T2-weighted sequences) and a pathologist reviewed the available histological material. PATIENTS: Seventeen patients with 18 histologically proven IM were reviewed. Histological samples of 11 of these 18 tumors were available for pathological analysis. RESULTS: There were 14 women and three men, with a mean age of 58.9 years. IM involved predominantly the thigh (n=10). MR imaging showed well-circumscribed intramuscular masses, hypointense on T1-weighted and hyperintense on T2-weighted images. Eleven masses were homogeneous and seven slightly heterogeneous due to fibrous septa. Enhanced MR imaging demonstrated three different patterns: peripheral enhancement (n=1), peripheral and patchy internal enhancement (n=7) or peripheral and linear internal enhancement (n=4). Intratumoral cysts were detected in four masses. MR imaging showed the presence of a pseudocapsule (n=12), fat around the lesion (n=16) and peritumoral edema (n=16). Histologically, all the tumors were hypocellular, hypovascular and myxoid. Peripheral areas of collagenous fibers formed a partial capsule and IM often merged into surrounding muscular fibers. More cellular tumors and those with scanty myxoid stroma tended to show a more prominent internal enhancement. CONCLUSION: IM shows several recognizable MR features which suggest its diagnosis. PMID- 15538561 TI - Compositional analysis of muscle in boys with Duchenne muscular dystrophy using MR imaging. AB - OBJECTIVE: Boys with Duchenne muscular dystrophy (DMD) present by age 5 years with weakness and, untreated, stop walking unaided by age 10 or 11 years. We used magnetic resonance (MR) imaging to study age-related changes in the composition and distribution of diseased muscles. DESIGN AND PATIENTS: Eleven boys (mean 7.1+/-1.6 years) with DMD underwent clinical and MR examinations. Quantitative muscle strength and timed functional testing was performed. Thigh muscles were scanned at three levels (hip, mid-thigh, and knee) using T1-weighted spin echo and short-tau inversion recovery (STIR) sequences. Outcome measures included intramuscular fatty infiltration, intermuscle fat deposition, edema, and muscle size. RESULTS: Ten boys completed the study. Older boys demonstrated more prominent fatty infiltration of muscles. Fatty infiltration occurred in a characteristic pattern with the gluteus and adductor magnus muscles most commonly involved and the gracilis most commonly spared. Similarly, patchy increases in free water content suggested a pattern of intramuscular edema or inflammation. Atrophy occurred in muscles heavily infiltrated with fat, and true hypertrophy selectively occurred in those that were spared. CONCLUSIONS: While fibrofatty changes have been described in DMD, this study further defines differential involvement and additionally suggests widespread edema or inflammation. Improved imaging techniques to quantify the degree and distribution of these changes may provide a basis for exploring mechanisms of action of medications and perhaps another means for selecting treatment regimens and monitoring their effects. PMID- 15538562 TI - Weismann-Netter-Stuhl syndrome in two siblings. AB - Cases of Weismann-Netter-Stuhl syndrome involving the upper extremities and affecting siblings have rarely been reported. We present the radiological findings of Weismann-Netter-Stuhl syndrome in two siblings, with upper extremity involvement in one of them. PMID- 15538564 TI - Functional outcome of burst fractures of the first lumbar vertebra managed surgically and conservatively. AB - We retrospectively reviewed 31 neurologically intact patients with burst L1 fractures. We obtained a follow-up clinical evaluation after a mean of 43 (14-80) months from 26 patients--11 treated surgically and 15 managed non-surgically. Patients were assessed with regard to pain, employment status, recreational activities and overall satisfaction. At final follow-up of 15 patients managed non-surgically, six had little or no pain; 12 had returned to work with six declaring little or no restrictions, and eight had returned to the same level of recreational activity as prior to injury with seven declaring little or no restrictions. Of 11 patients treated surgically, four had little or no pain; seven had returned to work with three declaring little or no restrictions, three had returned to the same level of recreational activity as prior to injury and four declared little or no restrictions. There was no correlation found between vertebral collapse, kyphosis, retropulsion and clinical outcome. Patients who had non-operative management reported a good functional outcome. However, patients who required surgical stabilisation due to different fracture characteristics reported a poorer functional outcome. PMID- 15538565 TI - Intraoperative localization of an osteoid-osteoma using a gamma probe. AB - The purpose of this study was to evaluate whether intraoperative nidus detection with a hand-held gamma probe was efficient enough for use as a routine procedure. Thirty-seven patients with osteoid-osteomas were submitted to surgical treatment. The first group consisted of 19 patients submitted to open nidus resection using a hand-held gamma probe as guide. The control group consisted of 18 patients operated on by conventional technique. The procedures were classified as successful if nidus resection could be confirmed by histology or postoperative imaging studies. Patients in the gamma group were followed for a mean of 13 months; patients in the control group for a mean of 39 months. In the gamma group, 17/19 procedures were successful; in the control group, only 12/18 procedures were successful. The gamma probe helped to locate the osteoid-osteoma nidus, and the same probe could be used in various hospitals. PMID- 15538566 TI - Refractory post-pancreatitis thrombotic thrombocytopenic purpura: response to rituximab. PMID- 15538567 TI - Distinction of acute lymphoblastic leukemia from acute myeloid leukemia through microarray-based DNA methylation analysis. AB - Patterns of DNA methylation are substantially altered in malignancies compared to normal tissue, with both genome-wide hypomethylation and regional increase of cytosine methylation at dinucleotides of cytosine and guanine, i.e., CpG dinucleotides. While genome-wide hypomethylation renders chromosomes instable, hypermethylation of CpGs in promoter regions is generally associated with transcriptional silencing, e.g., of tumor suppressor genes. To investigate whether disease-specific methylation profiles exist for different entities of acute leukemia, a microarray-based DNA methylation analysis simultaneously assessing 249 CpG dinucleotides originating from 57 genes was employed. Hereby, samples from precursor B-cell acute lymphoblastic leukemia (ALL) could be distinguished from cases of acute myeloid leukemia by virtue of N33, EGR4, CDC2, CCND2, or MOS hypermethylation in ALL. PMID- 15538568 TI - Long-term results of dose density therapy in patients with aggressive lymphoma. AB - To evaluate the long-term outcome of dose density chemotherapy in the treatment of aggressive lymphoma, we analyzed 142 patients with untreated aggressive lymphoma. Chemotherapy was an eight-drug regimen given in weekly intervals in two prospective trials. The median observation period was 8 years; the longest follow up was 13 years. Overall survival at 8 years was 0.583. The 8-year survival of patients < or =60 years was significantly better than that of older patients, namely 0.713 vs 0.304 (p=0.000000697). This excellent survival of patients aged < or =60 years was identical for high-risk and high-intermediate-risk patients compared with low-risk and low-intermediate-risk patients in the age-adjusted international prognostic index (IPI). The excellent long-term results of the CEOP/IMVP-Dexa regimen (cyclophosphamide, epirubicin, vincristine, and prednisone/ifosfamide with systemic mesna, methotrexate, etoposide, and dexamethasone) for patients aged < or =60 years suggest that this regimen might be superior to the standard CHOP regimen (cyclophosphamide, doxorubicin, vincristine, and prednisolone) and needs to be tested in comparison to high-dose regimens and novel approaches including antibody treatment. PMID- 15538569 TI - Quinoxaline 1,4-dioxides induce G2/M cell cycle arrest and apoptosis in human colon cancer cells. AB - We have recently shown that quinoxaline 1,4-dioxide (QdNO) derivatives, namely 2 benzoyl-3-phenyl-6,7-dichloroquinoxaline 1,4-dioxide (DCQ), 2-benzoyl-3-phenyl quinoxaline 1,4-dioxide (BPQ) and 2-acetyl-3-methyl-quinoxaline 1,4-dioxide (AMQ), suppress the growth of T-84 human colon cancer cells. Here we show that the growth-suppressive effects of QdNOs are due to their ability to induce cell cycle arrest and/or apoptosis. While AMQ blocked more than 60% of cells at the G2/M phase without inducing apoptosis, DCQ caused a significant increase in apoptotic cells with no noticeable effects on the cycling of cells. Treatment with BPQ resulted in G2/M cell cycle arrest and induction of apoptosis. With regard to the effects of QdNOs on molecules that regulate apoptosis and the G2 to M transition, both BPQ and AMQ inhibited the expression of cyclin B, while DCQ significantly decreased the levels of Bcl-2 and increased Bax expression. Next, we investigated whether transforming growth factor-beta1 (TGF-beta1) and/or extracellular signal-regulated kinase (ERK) mediate the antiproliferative and apoptotic effects of QdNOs in colon cancer cells. Interestingly, the above QdNOs increased differentially total TGF beta1 mRNA expression and decreased TGF alpha mRNA and ERK phosphorylation. None of these QdNOs induced changes in TGF beta-2 mRNA expression. The addition of a specific inhibitor of MEK greatly enhanced apoptosis in cells treated with DCQ, suggesting that the inhibition of ERK phosphorylation may explain, to an extent, the apoptogenic effects of this compound. Taken together, these findings provide insights into possible molecular mechanisms of growth inhibition by QdNOs that could aid in their evaluation for anticancer therapy. PMID- 15538570 TI - A phase IB clinical and pharmacokinetic study of the angiogenesis inhibitor SU5416 and paclitaxel in recurrent or metastatic carcinoma of the head and neck. AB - PURPOSE: SU5416 is a novel small organic molecule that non-competitively inhibits the phosphorylation of the VEGF tyrosine kinase receptor, Flk-1. This phase IB study was performed to determine the safety, pharmacokinetics, and preliminary efficacy of the combination of SU5416 and paclitaxel in recurrent or metastatic carcinoma of the head and neck. METHODS: Enrolled in the study were 12 patients with biopsy-proven recurrent or metastatic carcinoma of the head and neck. Six patients received intravenous SU5416 110 mg/m2 on days 1, 15, 18, 22 and 25, and paclitaxel 70 mg/m2 on days 8, 15 and 22. Since two patients experienced a dose limiting toxicity (DLT) in cohort 1, the next six patients received identical treatment as above except the paclitaxel dose was reduced to 55 mg/m2 per week. RESULTS: A total of 42 cycles at two different dose levels were given. In cohort 1 there were two deep venous thromboses that were DLTs. In the second cohort there was a DLT consisting of a transient ischemic attack after receiving SU5416. Most of the other toxicities seen were grade 1 or 2 in nature and consisted of headache, facial flushing, and fatigue. Two patients developed extensive ulcerative cavities at sites of prior radiation. There were no significant changes in the pharmacokinetic parameters of SU5416 given with paclitaxel. Four patients had prolonged freedom from progression of 18, 28, 42, and 60 weeks duration. CONCLUSIONS: The combination of SU5416 with paclitaxel had a higher than expected incidence of thromboembolic events and prophylactic anticoagulation should be considered for future trials that combine an angiogenesis inhibitor with cytotoxic chemotherapy. Although the future development of SU5416 as a chemotherapeutic agent is unclear, there was a clinical benefit seen with this combination in 36% of the patients. This trial supports the use of developing antiangiogenic combinations, using molecular targeted agents, in head and neck carcinoma. PMID- 15538571 TI - Apoptosis of murine melanoma B16-BL6 cells induced by quercetin targeting mitochondria, inhibiting expression of PKC-alpha and translocating PKC-delta. AB - PURPOSE: In our previous study, quercetin was found to induce apoptosis of murine melanoma B16-BL6 cells. The cellular and molecular mechanism of quercetin-induced apoptosis was investigated in the present study. METHODS: Nuclear morphology was determined by fluorescence microscopy. DNA fragmentation was analyzed by electrophoresis and quantified by the diphenylamine method. The transmembrane potential of mitochondria was measured by flow cytometry. Bcl-2, Bcl-X(L), PKC alpha, PKC-beta, and PKC-delta were detected by Western blotting. Caspase activity was determined spectrophotometrically. RESULTS: Quercetin induced the condensation of nuclei of B16-BL6 cells in a dose-dependent pattern as visualized by Hoechst 33258 and propidium iodide dying. Phorbol 12-myristate 13-acetate (PMA), a PKC activator, significantly enhanced apoptosis induced by quercetin, while doxorubicin, a PKC inhibitor, markedly decreased it. Both PMA and doxorubicin showed a consistent effect on the fragmentation of nuclear DNA caused by various dosages of quercetin. Quercetin dose-dependently led to loss of the mitochondrial membrane potential, which was also significantly reinforced or antagonized by PMA and doxorubicin, respectively. Moreover, PMA showed reinforcement, while doxorubicin showed significant antagonization, of the quercetin-mediated decrease in the expression of Bcl-2. Quercetin promoted caspase-3 activity in a dose-dependent manner, which was also regulated by PMA and doxorubicin with a pattern similar to that seen in their effect on apoptosis, mitochondrial membrane potential and Bcl-2 expression, but none of these were directly affected by PMA and doxorubicin. Free fatty acid and chlorpromazine, a PKC activator and inhibitor, respectively, did not interfere with these effects of quercetin. B16-BL6 cells expressed PKC-alpha, PKC-beta, and PKC-delta. Quercetin dose-dependently inhibited the expression of PKC-alpha but not that of PKC-beta and PKC-delta. Doxorubicin almost completely blocked the effect of quercetin on the expression of PKC-alpha. Quercetin was also involved in the translocation of PKC-delta from the cytosol to the nucleus. PMA enhanced the effect of quercetin on the translocation of PKC-delta. CONCLUSIONS: These results indicate that quercetin induced apoptosis of murine melanoma B16-BL6 cells by injuring their mitochondria, increasing the activity of caspase-3, inhibiting the expression of Bcl-2 and PKC-alpha, and inducing the translocation of PKC-delta. Doxorubicin inhibited these effects of quercetin by blocking the decreased expression of PKC-alpha induced by quercetin while PMA increased these effects by enhancing the translocation of PKC-delta induced by quercetin. PMID- 15538572 TI - [Postmortal diagnosis of Parkinson's disease]. AB - Parkinson's disease is a continuously progressive degenerative disorder of the central, peripheral and enteric human nervous systems. Not only the substantia nigra, but also a number of other components of the motor and limbic systems, as well as the autonomic regulation, suffer heavy damages. Only a few of the many types of nerve cells in the human central nervous system develop the characteristic Lewy bodies and Lewy neurites. They are composed primarily of aggregated alpha-synuclein and lead to the premature destruction of the affected neurons. Due to the selective neuronal vulnerability, a distinctive distribution of changes occurs within the central nervous system, leading to a corresponding loss of functionality in many systems. The changes occur in an ordered timely fashion. The ascending pathological process begins within the brain at the glossopharyngeal and vagal areas, nearly destroys the substantia nigra, and reaches the mesocortex of the gray matter. From here it expands to further areas of the neocortex, thereby marking the end phase of the disease. PMID- 15538573 TI - Variation in mitochondrial transcript profiles of protein-coding genes during early germination and seedling development in wheat. AB - We examined RNA profiles of wheat mitochondrial genes during the developmental period when seeds leave dormancy, germinate and develop into seedlings. Mitochondrial RNAs isolated from 0 h to 6 days post-imbibition were subjected to Northern analysis, using coding-specific and intron-specific probes. Stable, edited mRNAs were observed in dormant seeds and precursor RNAs were subsequently detected early in embryo germination. The respiratory chain genes (nad7, cox1, cox2, atp6) showed mRNA profiles which paralleled those of the ribosomal RNAs, whereas ribosomal protein genes (rps2, rps3, rps7) had proportionately lower steady-state mRNA levels in later stages of seedling development. The relative levels of precursors compared with the respective mRNAs shifted down during development, consistent with transcription outpacing RNA processing in the early stages but co-ordination being more effective several days after imbibition. In the case of multiply split genes containing group II introns, complex patterns of splicing intermediates were observed, suggesting a lack of strict polarity of intron removal, although splicing efficiency appears to differ among introns. Excised intron RNAs typically are relatively more abundant in embryos than seedlings. These observations are consistent with a transient imbalance of RNA processing machinery at the onset of seed germination, which is a period of rapid mitochondrial biogenesis. PMID- 15538574 TI - Ankylosing spondylitis and bone mineral density--what is the ideal tool for measurement? AB - Ankylosing spondylitis (AS) is characterised by chronic inflammation and partial ossification, yet vertebral fractures due to osteoporosis, although common, are frequently unrecognised. The aim of this study was to (1) show the frequency of changes in the progress of osteopenia/osteoporosis in AS depending on duration and stage of the disease and (2) assess the ranking of two different methods of bone density measurement in this clinical pattern. We measured bone density in 84 male and female patients with both dual X-ray absorptiometry (DXA) and single energy quantitative computed tomography (SE-QCT). In the initial and advanced stages of the disease, a high decrease in axial bone density could be verified (DXA: osteopenia in 5% and osteoporosis in 9.2%; SE-QCT: osteopenia in 11.8% and osteoporosis in 30.3%). Peripheral bone density decrease as in osteopenia could be proven in 17.6% by DXA measurement. With SE-QCT, a decrease in vertebral trabecular bone density could already be observed in the initial stage and continued steadily during the course of the disease; cortical bone displayed the same trend up to stages of ankylosis. With DXA, valid conclusions are more likely to be expected in less marked ankylosing stages of AS. In stages of advanced ankyloses in the vertebral region (substantial syndesmophytes), priority should be given to SE-QCT, due to the selective measurement of trabecular and cortical bone. The DXA method often yields values that are too high, and the replacement of vertebral trabecular bone by fatty bone marrow is not usually recorded as standard. There may already be an increased risk of bone fracture in AS in osteopenia on DXA along with an osteoporosis already established on SE-QCT. PMID- 15538575 TI - Agrobacterium-mediated transformation of the wetland monocot Typha latifolia L. (Broadleaf cattail). AB - An Agrobacterium-mediated model transformation system was standardized for the wetland monocot Typha latifolia L. to achieve the long-term objective of introducing candidate genes for phytoremediation. Two binary plasmid vectors, pCAMBIA1301/EHA105 and pTOK233/LBA4404, both containing the gus (beta glucuronidase) and hptII (hygromycin phosphotransferase II) genes, were used for transformation. Fifty-day-old 5 mg/l picloram-derived calli were cocultivated and selected on medium containing 20 mg/l or 40 mg/l hygromycin. Resistant calli were regenerated on medium supplemented with 5 mg/l 6-benzylaminopurine, with or without 20 mg/l or 40 mg/l hygromycin and with or without charcoal (10 g/l). Transient GUS activity in explants ranged between 28% and 36%. Hygromycin resistant calli, selected after 3 months, showed stable GUS expression. A total of 46 plants were regenerated and established in the greenhouse; 13 showed stable GUS expression. Cocultivation of dark culture-derived calli, directly selected on regeneration medium containing 20 mg/l hygromycin and rooted on medium with 20 mg/l hygromycin was the best protocol. The addition of charcoal did not have any effect on regeneration. PCR and Southern analyses of transgenic calli and transgenic plants confirmed the presence of the introduced genes. In conclusion, T. latifolia could be genetically transformed by Agrobacterium tumefaciens. PMID- 15538576 TI - Biolistic transformation of highly regenerative sugar beet (Beta vulgaris L.) leaves. AB - Leaves of greenhouse-grown sugar beet (Beta vulgaris L.) plants that were first screened for high regeneration potential were transformed via particle bombardment with the uidA gene fused to the osmotin or proteinase inhibitor II gene promoter. Stably transformed calli were recovered as early as 7 weeks after bombardment and GUS-positive shoots regenerated 3 months after bombardment. The efficiency of transformation ranged from 0.9% to 3.7%, and stable integration of the uidA gene into the genome was confirmed by Southern blot analysis. The main advantages of direct bombardment of leaves to regenerate transformed sugar beet include (1) a readily available source of highly regenerative target tissue, (2) minimal tissue culture manipulation before and after bombardment, and (3) the overall rapid regeneration of transgenic shoots. PMID- 15538577 TI - Analysis of transcripts in methyl jasmonate-treated ginseng hairy roots to identify genes involved in the biosynthesis of ginsenosides and other secondary metabolites. AB - Methyl jasmonate (MeJA) treatment increases the levels of plant secondary metabolites, including ginsenosides, which are considered to be the main active compounds in ginseng (Panax ginseng C.A. Meyer). To create a ginseng gene resource that contains the genes involved in the biosynthesis of secondary metabolites, including ginsenosides, we generated 3,134 expression sequence tags (ESTs) from MeJA-treated ginseng hairy roots. These ESTs assembled into 370 clusters and 1,680 singletons. Genes yielding highly abundant transcripts were those encoding proteins involved in fatty acid desaturation, the defense response, and the biosynthesis of secondary metabolites. Analysis of the latter group revealed a number of genes that may be involved in the biosynthesis of ginsenosides, namely, oxidosqualene cyclase (OSC), cytochrome P450, and glycosyltransferase. A novel OSC gene was also identified by this analysis. RNA gel blot analysis confirmed that transcription of this OSC gene, along with squalene synthase (SS) and squalene epoxidase (SE) gene transcription, is increased by MeJA treatment. This ginseng EST data set will also provide important information on the genes that are involved in the biosynthesis of other secondary metabolites and the genes that are responsive to MeJA treatment. PMID- 15538579 TI - Uterine artery embolisation for uterine fibroids using a 4F Rosch inferior mesenteric catheter. AB - PURPOSE: To evaluate in a prospective study the use of a 4F Rosch inferior mesenteric (RIM) catheter for uterine artery embolisation (UAE). MATERIALS AND METHODS: UAE was performed in 72 women over a 37-month period. A 4F RIM braided J curve 65-cm catheter was used in combination with an angled hydrophilic 150 cm, 0.035" flexible tip guide-wire to catheterise the horizontal portion of both uterine arteries (UA) from a right common femoral artery (CFA) approach. Technical success was defined as successful catheterisation and embolisation of both uterine arteries. Fluorosocpic and procedure times were recorded. RESULTS: Mean subject age=43.7 years (range=25-57 years). Technical success was 98.6% (n=71/72). A single approach via the right CFA was used in 88.9% (n=64/72) and a bilateral CFA approach in 11.1% (n=8/72). Bilateral uterine artery catheterisation using a single 4F RIM catheter via the right CFA approach was successful in 79.2% (n=57/72). Microcatheters were used in 2.8% of patients (n=2/72). Mean fluoroscopic time=13.6+/-5.3 min (mean+/-SD). Mean procedure time=44.2+/-16.5 min. CONCLUSION: High technical success rate for UAE is possible using a single 4F RIM catheter via a unilateral right CFA approach, which obviates the need for Waltman loop formation, reversed curve catheters and complex suture-catheter arrangements. PMID- 15538578 TI - Apparent diffusion coefficient in cervical cancer of the uterus: comparison with the normal uterine cervix. AB - A relation between apparent diffusion coefficient (ADC) values and tumor cellular density has been reported. The purpose of this study was to measure the ADC values of cervical cancers in the uterus and compare them with those of normal cervical tissues, and to test whether ADC could differentiate between normal and malignant cervical tissues in the uterus. Twelve consecutive female patients with cervical cancer of the uterus and ten female patients with other pelvic abnormalities were included in this study. ADC was measured at 1.5 T with b factors of 0, 300 and 600 s/mm2 using single-shot echo-planar diffusion-weighted imaging and a parallel imaging technique. The mean ADC value of cervical cancer lesions was 1.09+/-0.20 x 10(-3) mm2/s, and that of normal cervix tissue was 1.79+/-0.24 x 10(-3) mm2/s (P<0.0001). In nine patients treated by chemotherapy and/or radiation therapy, the mean ADC value of the cervical cancer lesion increased significantly after therapy (P<0.001). The present study showed, with a small number of patients, that ADC measurement has a potential ability to differentiate between normal and cancerous tissue in the uterine cervix. Further study is necessary to determine the accuracy of ADC measurement in monitoring the treatment response. PMID- 15538580 TI - Spinal vascular malformations. AB - Spinal vascular malformations are rare diseases that consist of true inborn cavernomas and arteriovenous malformations (including perimedullary fistulae, glomerular and juvenile AVMs) and presumably acquired dural arteriovenous fistulae. This review article gives an overview of the imaging features both on MRI and angiography, the differential diagnoses, the clinical symptomatology and the potential therapeutic approaches to these diseases. It is concluded that MRI is the diagnostic modality of first choice in suspected spinal vascular malformation and should be complemented by selective spinal angiography. Treatment in symptomatic patients offers an improvement in the prognosis, but should be performed in specialized centers. Patients with spinal cord cavernomas and perimedullary fistulae type I are surgical candidates. Dural arteriovenous fistulae can either be operated upon or can be treated by an endovascular approach, the former being a simple, quick and secure approach to obliterate the fistula, while the latter is technically demanding. In spinal arteriovenous malformations, the endovascular approach is the method of first choice; in selected cases, a combined therapy might be sensible. PMID- 15538581 TI - Mastoid effusion associated with dural sinus thrombosis. AB - We present a series of three patients with mastoid air cell effusions associated with adjacent lateral sinus thrombosis. In all of these cases, the findings support the hypothesis that the mastoid effusion is secondary to sinus thrombosis rather then the other way around. Also shown is the chronology and natural evolution of mastoid air cell effusion secondary to sinus thrombosis as seen on planar imaging. PMID- 15538582 TI - Short-term ureteral catheter stenting after uncomplicated transurethral uretero lithotomy. AB - Short-term (1 or 2 post-operative days) ureteral catheter stenting after transurethral uretero-lithotomy (TUL) to avoid flank pain due to transient ureteral edema is described. Patients who underwent TUL for middle or distal ureteral stones with a rigid ureteroscope without complications during the procedures were the candidates for short-term ureteral stenting. An end-hole ureteral catheter, used to insert a guide wire during TUL, were used for stenting. The tip of the catheter was located near the renal pelvis and the other end was introduced outside through the urethra with a 14 F urethral catheter. The stent and catheter were removed on post-operative day 1 or 2. For the 18 patients treated using this method, the time of analgesic use after stent removal was 0.6+/-0.8, indicating a sufficient duration of stenting. Short-term ureteral catheter stenting is a cheap and easy way for post-operative management for uncomplicated TUL. PMID- 15538583 TI - [Solitary iris metastasis from breast cancer. Effective local therapy with electron beam irradiation]. AB - BACKGROUND: The prevalence of intraocular metastases from breast cancer is approximately 4-5%. Solitary metastases of the iris are rare. We report on successful treatment of a solitary iris metastasis using electron beam irradiation. CASE REPORT: A 30-year-old patient presented with an amelanotic tumor of the iris and the anterior chamber angle of her right amblyopic eye. The patient had undergone left-sided breast-conserving surgery and lymph node dissection 3 years before followed by chemotherapy and radiotherapy. The iris tumor was considered a metastasis. Fractionated electron beam irradiation was performed applying a total dose of 50 Gy in fractions of 5 x 2 Gy/week, electrons (9 MeV). The iris metastasis was completely resolved 13 months after radiotherapy. Until now no signs of cataract have been detected and visual acuity has remained stable. CONCLUSION: Electron beam irradiation of this iris metastasis was an effective treatment for preserving visual acuity and ocular function with tolerable acute toxicity and so far no adverse side effects. PMID- 15538584 TI - [Neurotrophic keratitis]. AB - Neurotrophic keratitis is a degenerative disease of the cornea caused by reduced corneal innervation. Trauma, tumors, inflammatory lesions and surgical procedures can damage the first branch of the trigeminal nerve on its entire course from brainstem to and within the cornea. Loss or reduction of corneal innervation leads to a reduced aqueous phase of the tear film and due to reduced supply with neurotransmitters/trophic factors also to reduced epithelial healing capacity (impaired mitosis and migration). Combined existence of tear film deficiency and impaired epithelial healing capacity predispose to persistent epithelial defects, corneal ulcers and perforation. Early diagnosis and adequate treatment may prevent this catastrophic chain of events. PMID- 15538585 TI - Sex differences in metabolic rates in field crickets and their dipteran parasitoids. AB - Sex differences in metabolic rate (MR) can result from dimorphism in the performance of energetically demanding activities. Male crickets (Teleogryllus oceanicus) engage in costly calling and aggressive activity not performed by females. Consistent with this difference, we found higher maximal MR, factorial scope, and fat content in males than females. T. oceanicus song is also costly because it attracts the parasitoid fly Ormia ochracea. Parasitized crickets had reduced maximal MR consistent with a metabolic cost to harboring larvae. This cost was greater for females, either because females invest more heavily into reproduction at the expense of metabolic capacity, or because males are under stronger selection to respond to infection. Little is known about O. ochracea outside of its auditory system and parasitic lifestyle. We observed greater resting MR in male flies, possibly reflecting a sex difference in the requirement for metabolic power output, because male flies perform potentially costly mating behavior not seen in females. We found a positive relationship between larval density within a cricket and pupal resting MR, suggesting that crickets in good condition are able to both harbor more larvae and produce larvae with higher resting MR. These results reveal a complex interplay between the metabolism of crickets and their fly parasitoids. PMID- 15538586 TI - Single-system ectopic ureters associated with renal dysplasia. AB - The purpose of this study was to inquire into the clinical features and methods for the diagnosis and therapy of single-system ectopic ureters associated with renal dysplasia. Intravenous urography (IVP), diuretic B-mode ultrasound (B-US), and, in four cases, computerized tomography (CT) were performed in twelve female patients with renal dysplasia-seven on the left and five on the right-and the clinical records were retrospectively summarized and analyzed. The main presentation was droplet urinary incontinence. IVP and B-US showed that in all in whom the affected kidneys did not function, the contralateral kidneys had undergone compensatory hypertrophy. Only in four cases were the dysplastic renal tissues displayed in the pelvic cavity on B-US. In eight cases, the affected kidneys were not found. Of the four patients who had CT scanning, the dysplastic kidneys were explored in the pelvis in two cases. All patients underwent ureteronephrectomy, and the urinary incontinence disappeared after operation. Single-system ectopic ureters associated with renal dysplasia mainly affect female children, and most present with droplet urinary incontinence; the affected kidney may not be displayed on ultrasonography and IVP, but the contralateral kidney may display compensatory hypertrophy on IVP and can easily be misdiagnosed as a solitary kidney. The curative effect of ureteronephrectomy is definite. PMID- 15538587 TI - A standardised protocol for the acute management of corrosive ingestion in children. AB - Oesophageal strictures developing after caustic ingestion in children are a serious problem, and several protocols to prevent stricture formation have been proposed. A prospective clinical trial was conducted for preventing strictures in caustic oesophageal burns in a single clinic, and the results are presented. All children with caustic ingestion who had oesophagoscopy for diagnosing the severity of the burn were included in the study. Eighty-one children were included in the series, with ages ranging between 3 months and 12 years. The patients were given nothing by mouth until oesophagoscopy. IV fluids, broad spectrum antibiotics, ranitidine, and a single-dose steroid were given. Oral burns were positive in 66 patients. Oesophagoscopy revealed a normal oesophagus in nine patients, grade 1 burn in 24, grade 2a in 21, grade 2b in 23, grade 3a in two, and grade 3b in one. Patients with grade 1 and 2a burns were discharged after oesophagoscopy. Patients with grade 2b and all grade 3 burns were given nothing by mouth for a week except water when swallowing their saliva, and were fed via total parenteral nutrition. After the 1st week, if there was no problem with swallowing, liquid foods were introduced. No intraluminal tubes were used. At the end of the 3rd week, a barium meal was administered and an upper gastrointestinal series taken. Dilatation was performed at 2-week intervals for strictures, which developed in one grade 2a patient, six grade 2b patients, and the grade 3b patient. Only one of these patients is currently on an oesophageal dilatation program. Limiting oral intake and avoiding foreign bodies in the oesophagus seem to provide a good success rate; however, further prospective studies are needed to decrease the incidence of corrosive oesophageal strictures. PMID- 15538588 TI - Hepatoblastoma in low birth weight infants: an institutional review. AB - The association between hepatoblastoma and low birth weight documented recently in the literature has yet to be well explained, in particular the suggestion that these patients may have a more aggressive form of the disease. From 1989 to 2003, our institution treated four patients for hepatoblastoma who had birth weights of less than 1,500 g. Notable was 100% patient survival despite bilateral and, in one case, recurrent disease. Speculation regarding the etiology of this subset of hepatoblastoma has included damage to developing hepatocytes induced by oxygen free radicals. Our patients universally had pulmonary disease requiring prolonged supplemental oxygen and ventilatory support. However, our review supports no changes in the standard care of low birth weight infants or in managing those who develop hepatoblastoma. PMID- 15538589 TI - Intraoperative cutting errors in total knee arthroplasty. AB - INTRODUCTION: Precise reconstruction of leg alignment offers the best opportunity for achieving good long-term results in total knee arthroplasty (TKA). It was the aim of this study to evaluate the bone-cutting process as a potential source of inaccuracy in TKA. MATERIALS AND METHODS: In a consecutive series of 50 computer assisted TKAs, cutting errors, which were defined as a difference between the cutting block position before sawing and the achieved resection plane afterwards, were measured for the distal femur and proximal tibia resection. Measurements were performed using a CT-based navigation system. RESULTS: For the distal femoral cut, there was a mean varus/valgus deviation of 0.6 degrees (SD+/-0.5 degrees ) and a mean flexion/extension deviation of 1.4 degrees (SD+/-1.3 degrees ). For the proximal tibia, varus/valgus alignment showed a mean deviation of 0.5 degrees (SD+/-0.5 degrees ). The mean sagittal variability was 1.0 degrees (SD+/ 0.9 degrees ). Differences between the frontal and the sagittal plane were significant. CONCLUSION: To minimize cutting errors, techniques and instruments are needed which enable a more stable fixation of the cutting blocks or even more appropriate preparation instruments. Using a computer-assisted technique, the surgeon is aware of cutting errors occurring at each point of the operation and will therefore be able to correct these errors during surgery, while he is not aware of those errors with the conventional TKA technique. PMID- 15538591 TI - Clinical outcome in patients with obsessive-compulsive disorder after discontinuation of SRI treatment: results from a two-year follow-up. AB - BACKGROUND: Combined treatment with serotonin-reuptake inhibitors (SRI) and cognitive-behavioral therapy (CBT) is a common therapy approach for obsessive compulsive disorder (OCD). However, it is a matter of debate whether discontinuation of SRI after combined treatment leads to relapse. METHOD: Seventy four consecutively admitted patients suffering from OCD were included in the study. Thirty-seven patients were treated with CBT alone, and 37 patients received combined CBT and SRI treatment. Of these latter patients, seventeen discontinued SRI treatment during the follow-up period (1 and 2 years after inpatient treatment). OCD symptom severity was determined by Yale-Brown Obsessive Compulsive Scale (Y-BOCS), and mood was assessed by Hamilton Depression Rating Scale (HDRS). RESULTS: During the initial treatment, scores for Y-BOCS (p < 0.001), HDRS (p < 0.001) and the Global Assessment of Functioning Scale (GAF) (p < 0.001) improved significantly in all groups. Reassessment two years later revealed that a) OCD symptom severity and depression scores were similar between the groups and b) discontinuation of SRI did not prompt by a recurrence of symptoms. CONCLUSIONS: We interpret our results as suggesting that discontinuation of SRI treatment may be considered in formerly combined treated OCD patients after stable remission. PMID- 15538590 TI - Premenstrual dysphoric symptoms amongst Brazilian college students: factor structure and methodological appraisal. AB - OBJECTIVES: The present study aims to assess the factor structure of the DSM-IV Premenstrual Dysphoric Disorder (PMDD) symptoms and its relationship with depressive symptoms. METHODS: We evaluated retrospectively PMDD symptoms in 513 female college students, through a self-reporting questionnaire based on DSM-IV criteria, in addition to the Beck Depression Inventory (BDI). Principal component analysis on PMDD symptom data was performed to assess its dimensional structure. RESULTS: In this non-clinical sample, the analysis indicated a higher importance of the dysphoric dimension, but physical symptoms as well as "being out of control" or "overwhelmed" should also be viewed as major symptoms of PMDD. Behavioural symptoms are of secondary importance. The mean BDI score of PMDD group was significantly higher (p < 0.05) than non-PMDD group. CONCLUSION: The factor structure of the total sample was similar to the symptom structure suggested by DSM-IV diagnostic criteria. Depressive symptoms should be viewed as a confounding variable in PMDD. PMID- 15538592 TI - Hypericum extract LI 160 and fluoxetine in mild to moderate depression: a randomized, placebo-controlled multi-center study in outpatients. AB - Efficacy and tolerability of Hypericum LI 160 was compared to fluoxetine and placebo in mild to moderate Major Depression (DSM-IV) in a 4-week randomized, double-blind trial. One hundred and sixty-three outpatients from 15 general practitioner centers received either 900 mg Hypericum LI 160, 20 mg fluoxetine, or placebo daily. Amelioration was measured by the Hamilton and the Montgomery Asberg Depression scales. Response and remission rates and global ratings by investigators and patients were measured. Adverse event reports, laboratory screening, vital signs, physical exams and ECG were collected. No significant differences could be observed regarding efficacy measures except for remission rate (Hypericum 24%; fluoxetine 28%; placebo 7 %). Hypericum was significantly better tolerated than fluoxetine. Hypericum LI 160 or fluoxetine were not more effective in short-term treatment in mild to moderate depression than placebo. PMID- 15538593 TI - Levodopa reverses gait asymmetries related to anhedonia and magical ideation. AB - Animals and men turn preferentially away from the hemisphere with the more active dopamine (DA) system. Consistent with the idea of a right-hemispheric hyperdopaminergia in schizophrenia, a leftsided turning bias was described for unmedicated psychotic patients. We investigated the modulating role of DA and schizophrenia-like thought on whole-body turns in a controlled double-blind study. The number of veers to either side when walking blindfolded straight ahead (20 meter) was assessed in 40 healthy righthanded men (20 men received levodopa, the remaining participants placebo). Side preferences were analyzed in terms of individuals' positive (Magical Ideation, MI) and negative (Physical Anhedonia, PhysAn) schizotypal features. In the placebo group, increasing MI scores were related to increasing left-sided veering and increasing PhysAn scores were related to increasing right-sided veering. In the levodopa group, this relationship between preferred veering side and type of schizotypy was reversed. The finding in the placebo group suggests an association between MI and a relative right-hemispheric hyperdopaminergia. Unexpectedly, levodopa did not enhance this veering bias, but reversed it, suggesting that psychosis-protective mechanisms exist in the healthy positive "schizotypic" brain. Also unexpectedly, levodopa made "anhedonics" veer like "magics" after placebo, suggesting that DA agonists suppress negative schizotypal symptoms. PMID- 15538594 TI - EEG-mapping differences between narcolepsy patients and controls and subsequent double-blind, placebo-controlled studies with modafinil. AB - The aim of the present study was to investigate the role of EEG mapping as an objective and quantitative measure of vigilance in untreated and modafinil treated narcoleptics, and compare it with the conventional neurophysiological method of the Multiple Sleep Latency Test (MSLT) and the subjective Epworth Sleepiness Scale (ESS). In 16 drug-free narcoleptics and 16 normal controls a baseline 3-min vigilance-controlled EEG (V-EEG) and a 4-min resting EEG (R-EEG) were recorded during midmorning hours. Thereafter, in a double-blind, placebo controlled crossover design, patients were treated with a 3-week fixed titration of modafinil (200, 300, 400 mg) and placebo. EEG-mapping, MSLT and ESS measures were obtained before and at the end of the third week of therapy. Statistical overall analysis by means of the omnibus significance test demonstrated significant EEG differences between untreated patients and controls in the resting condition only (R-EEG). Subsequent univariate analysis revealed an increase in absolute and relative theta power, a decrease in alpha-2 and beta power as well as a slowing of the dominant frequency and the centroids of the alpha, beta and total power spectrum and thus objectified a vigilance decrement in narcolepsy. Modafinil 400 mg/d significantly improved vigilance as compared with placebo (p < or = 0.01), inducing changes opposite to the aforementioned baseline differences (key-lock principle). The MSLT and the ESS also improved under modafinil as compared with placebo, but changes were less consistent. Spearman rank correlations revealed the highest correlations between EEG mapping and the ESS, followed by those between EEG mapping and the MSLT, while the lowest correlation was found between the MSLT and the ESS. In conclusion, EEG mapping is a valuable instrument for measuring vigilance decrements in narcolepsy and their improvement under psychostimulant treatment. PMID- 15538595 TI - Assessing dimensional and categorical aspects of depression: validation of the AMDP Depression Scale. AB - BACKGROUND: A new observer depression scale which is based on the descriptive operationalized classification principles of ICD-10 is introduced. The AMDP Depression Scale (AMDP-DS), a 22-item clinician-rated inventory, provides the opportunity of dimensional as well as categorical depression severity assessment. METHODS: A sample of 50 patients with depressive symptoms were assessed with the AMDP-DS by two raters in a joint rater session. A number of widely used depression scales were applied separately by an independent rater. The ICD-10 symptom checklist was used for categorical assessment of depressive disorders. RESULTS: The inter-rater reliability for the total score of the AMDP-DS was excellent (Intraclass coefficient: 0.97). There were high correlations with the sum scores of the other scales under study. The correlations between the ICD-10 symptom-checklist and the AMDP-DS ranged from 0.75 to 0.89 for the total sample and 0.44 to 0.51 for patients with acute depressive episode. LIMITATIONS: The results are limited by the method of a joint rater setting and the relatively small subsample of patients with depressive episode. CONCLUSIONS: The AMDP-DS satisfies the requirements of psychometric criteria with a sufficient degree. A new depression scale designed to cover dimensional as well as categorical aspects has successfully been developed. PMID- 15538596 TI - Low doses of clozapine may stabilize treatment-resistant bipolar patients. AB - Open, uncontrolled studies suggest clozapine can have mood-stabilizing effects in treatment-resistant bipolar disorder. Unfortunately, the side effect profile limits clozapine's use at high doses. We report a series of nine bipolar I disorder patients who improved on relatively low doses of clozapine add-on therapy (250 mg or lower). Retrospectively abstracted clinical data identified nine patients with bipolar I disorder, as defined by DSMIV criteria, treated with low-dose clozapine at inpatient and outpatient settings. Monthly symptom evaluations were collected prospectively using standard assessments. Symptoms of mania and mood lability improved in all patients. Three patients demonstrated striking mood stabilization and returned to previous levels of functioning; five patients evidenced moderate improvement in mood stabilization and functioning; and one patient showed a minimal response. Overall, clozapine did not have a significant antidepressant effect. The mean clozapine dose at the end of the study was 156.3 +/- 77.6 mg/day, and duration of treatment was 12 months. Residual side effects were mild. The symptomatic improvement in these prospectively evaluated patients is consistent with our clinical impression in the majority of patients with bipolar disorder taking clozapine. PMID- 15538597 TI - Olfactory deficits in anorexia nervosa. AB - OBJECTIVE: Young patients admitted to the hospital due to anorexia nervosa report reduced pleasure and impaired perception of smell while eating. So far, two studies on odour identification ability in eating disorders did not suggest any significant deficits. Therefore a new and more detailed method of olfactory testing may be needed, in order to determine the subjective impairment of olfaction. METHOD: By using all three subtests of the recently developed smell test called "Sniffin'Sticks", the olfactory deficits were assessed in more detail and the results of female anorectic patients (n = 17) were compared with those of healthy females (n = 15). RESULTS: By examining the anorectic patients no deficits in the subtest odour identification were found. On the contrary, in the subtests odour discrimination and odour threshold deficits of the anorectic patients were detected. CONCLUSION: Reduced olfactory perception might be considered as a common deficit in anorexia nervosa with possible influences and consequences for therapy. PMID- 15538598 TI - Prepulse inhibition of the acoustically evoked startle reflex in patients with an acute schizophrenic psychosis--a longitudinal study. AB - Deficits in sensorimotor gating as assessed by prepulse inhibition (PPI) of the startle reflex have been reported in schizophrenia. However, the state or trait nature of these deficits and the relationships with clinical features and psychopathological symptoms are not clear. To explore these issues, we performed a longitudinal study with schizophrenia inpatients. We examined 36 medicated schizophrenia inpatients twice in the course of an acute psychotic episode: recently after admission and after psychopathological improvement 2-3 weeks later. In addition, we examined 18 healthy control subjects twice (two weeks apart). Relative to control subjects, patients with schizophrenia had lower PPI only in the acute, but not in the improved clinical state. Larger PPI deficits were associated with more severe formal thought disorder and bizarre behavior. In the present longitudinal study, PPI deficits in schizophrenic patients appeared to be state dependent. Taking into account recent evidence from the literature we propose that reduced PPI may be a mediating vulnerability marker of schizophrenia: Impairments in sensorimotor mechanisms which subserve PPI of the startle reflex may both predispose individuals to develop psychosis, and, in addition, may covary with the presence of acute positive symptoms. PMID- 15538599 TI - Structural brain differences in patients with schizophrenia and schizotypal disorder demonstrated by voxel-based morphometry. AB - Brain abnormalities of schizophrenia probably consist of deviation related to the vulnerability and pathological changes in association with overt psychosis. We conducted a cross-sectional comparison in brain morphology between patients with overt schizophrenia and schizotypal disorder, a schizophrenia-spectrum disorder without florid psychotic episode. Voxelbased morphometry was applied to assess gray matter volume in 25 patients with schizophrenia, 25 patients with schizotypal disorder, and 50 healthy control subjects. In comparison with controls, schizophrenia patients showed gray matter reductions in the bilateral medial frontal, inferior frontal, medial temporal, and septal regions, and the left middle frontal, orbitofrontal, insula, and superior temporal regions, and an increased gray matter in the left basal ganglia. Schizotypal disorder patients showed reductions in the left inferior frontal, insula, superior temporal, and medial temporal regions. There was a significant reduction in the left orbitofrontal region of schizophrenia compared with schizotypal disorder. Gray matter reductions that are common to both patient groups such as those in the left medial temporal and inferior frontal regions may represent vulnerability to schizophrenia, and additional involvement of several frontal regions may be crucial to florid psychosis. PMID- 15538600 TI - Early traumatic life events, parental rearing styles, family history of mental disorders, and birth risk factors in patients with social anxiety disorder. AB - OBJECTIVE: Childhood traumatic experiences, rearing styles, familial mental disorders and birth risk factors have been associated with the development of social anxiety disorder. METHOD: Patients with social anxiety disorder (n=50) and healthy controls (n=120) were investigated using a retrospective interview with 203 questions. RESULTS: The frequency of reports of traumatic childhood experiences was significantly higher in patients than in controls, including separation from parents, parents' marital discord, sexual abuse, familial violence, childhood illness, and other factors. On a 0-10 point 'severe trauma scale' patients had significantly higher mean scores (2.0; SD 1.28) than control subjects (0.82; SD 1.1; p<0.0001). Only 6 (12%) of the social phobic patients, but 63 (52.5%) of the controls did not report any severe traumatic events at all (chi2=24.0; p<0.0001). Compared to controls, patients described their parents' rearing styles as significantly more unfavourable. Patients reported higher rates of psychiatric disorders in their families in general, in particular anxiety disorders, depression, and suicidality. Birth risk factors did not differ between patients and controls. In a logistic regression model, the highest contribution was noted for familial anxiety disorders. Separation from parents also had a significant, but smaller influence. There was only a trend towards a significant contribution of childhood sexual abuse. Violence in the family, parental rearing styles and birth risk factors did not contribute significantly. CONCLUSIONS: The present data suggest that the aetiology of social anxiety disorder is multifactorial and that familial mental disorders and separation experiences are the most important contributing factors. PMID- 15538601 TI - SPECT imaging of serotonin transporter binding in patients with generalized anxiety disorder. AB - The purpose of this study was to characterize the binding properties of serotonin transporter (5-HTT) in the brain of the patients with generalized anxiety disorder (GAD) in comparison to healthy subjects using single photon emission computer tomography (SPECT) with the radioligand [123I]nor-beta-CIT. The subjects were 7 patients with GAD and 7 matched healthy volunteers. The regions of interest (ROI) were the midbrain and the thalamus. The comparison of the volumes of distribution did not show significant differences between the patients and controls in the binding of nor-beta-CIT to 5-HTT in the ROI. Binding of 5-HTT in the midbrain of patients was significantly and negatively correlated with their anxiety levels measured by the visual analogue scale immediately before the first scan (r=-0.79, p=0.035). This study failed to demonstrate an altered functional activity of 5-HTT in patients with GAD when compared with controls. PMID- 15538602 TI - Increased risk of developing stroke for patients with major affective disorder--a registry study. AB - Only a few studies have evaluated depressive disorder as a risk factor for cerebrovascular disease. In a hospital discharge register with nation-wide coverage of all hospitals in Denmark we used linkage between the somatic and psychiatric registries to study comorbidity between affective disorders and cerebrovascular diseases in hospitalised patients. The main finding of this study was that patients with depression severe enough to be hospitalised, seem to be at an increased risk of developing cerebrovascular disease. The hazard ratio of getting a diagnosis of stroke after initially having been discharged with a diagnosis of depression was found to be 1.22 (95% Confidence Interval: 1.06 1.41). In the group of patients with manic/bipolar disorder no association was found concerning development of stroke. In elderly with first time depression admitted to hospital, neurological disorders should be carefully evaluated and especially the risk of stroke should be considered. PMID- 15538603 TI - Platelet monoamine oxidase activity and psychometric correlates in male violent offenders imprisoned for homicide or other violent acts. AB - Violent behavior has been associated with certain personality traits like poor impulse control, sensation seeking, and monotony avoidance, which predispose to increased risk for violent acts. Low platelet monoamine oxidase (MAO) activity has been found in subjects with such traits, as well as in imprisoned violent offenders. On the other hand, cigarette smoking is expected to inhibit MAO activity. We assessed impulse control, lifetime history of aggression, suicide risk, as well as platelet MAO activity in 82 male offenders (mean age 33.0+/-10.7 years) imprisoned for homicide or other violent acts, and in 54 control male subjects (mean age 35.1+/-6.6 years). General psychopathology was assessed by the Hopkins Symptom Check List (SCL-90) questionnaire. A high rate of smoking (89%),significantly higher than controls (63%), was observed in offenders, but there were no differences in MAO activities among nonsmokers, moderate, or heavy smokers in either group. Offenders showed significantly higher scores in Impulse Control, Past Feelings and Acts of Violence, Suicide Risk and SCL-90 scales. Offenders who had committed other violent acts than homicide had higher scores in the three psychometric scales and in the Depression, Obsessive-Compulsive, Hostility, and Anxiety subscales of the SCL-90, than offenders who had committed homicide. Platelet MAO activity was significantly lower (p=0.01) in the offenders' group (38.1+/-14.4) compared to controls (44.7+/-15.2). The difference could not be attributed to smoking, and seems to be related to personality traits and behaviors that characterize the offenders' group. PMID- 15538604 TI - Electroconvulsive therapy for the treatment of clozapine nonresponders suffering from schizophrenia--an open label study. AB - OBJECTIVE: This open label study describes the efficacy of electroconvulsive therapy (ECT) as adjunctive treatment in clozapine nonresponders suffering from schizophrenia. METHOD: The results of clozapine and ECT treatment in 11 clozapine nonresponders suffering from schizophrenia are reported in terms of remission and relapse. RESULTS: Eight patients had a remission with this combination treatment. After remission of symptoms five patients had a relapse. Three of the five patients who relapsed had a second successful ECT course and remained well with maintenance ECT and clozapine. No evidence for adverse effects was found. CONCLUSION: Adjunctive ECT can be efficacious in clozapine nonresponders suffering from schizophrenia. PMID- 15538605 TI - Prevalence of attention deficit-/hyperactivity disorder (ADHD) and comorbid disorders in young male prison inmates. AB - OBJECTIVE: This study was performed to evaluate the prevalence of ADHD as well as comorbid conditions among young male prison inmates. METHOD: We investigated 129 prison inmates (mean age+/-SE: 19.2+/-2.0 years) and 54 healthy male control subjects (mean age+/-SE: 22.2+/-3.12 years) for the presence of adult ADHD using the Wender Utah Rating Scale (WURS), the Eysenck Impulsivity Questionnaire (EIQ), the diagnostic criteria for ADHD according to DSM-IV and ICD-10-research criteria and the Utah criteria for adult ADHD. In order to determine comorbid personality disorders we applied the International Personality Disorder Examination (IPDE). Externalization (ED) and Internalization Disorders (ID) were evaluated by means of the Achenbach Scales. Alcoholism (ALC) was examined via the Alcohol Use Disorder Test (AUDIT) and substance use disorder (SUD) has been investigated with the Structured Clinical Interview for DSM-IV (SCID). RESULTS: The overall prevalence of ADHD according to DSM-IV was 45%. The prevalence of disturbance of activity and attention (DAA) and hyperkinetic conduct disorder (HCD) via the ICD 10 research criteria was 21.7%. Sole DAA without any comorbid condition could be detected in one case. The most common diagnostic combinations were DAA/HCD and SUD/ALC (89% of all DAA/HCD cases). CONCLUSION: The prevalence of DAA/HCD or ADHD in young adult prison inmates is significantly elevated when compared to nondelinquent controls. Generally the population of young adult male prison inmates exhibits a considerable psychiatric morbidity. Of the total sample, 64% suffered from at least 2 disorders. Only 8.5% had no psychiatric diagnoses. This indicates the urgent need for more psychiatric expertise in young offender facilities. PMID- 15538606 TI - Charles Bonnet syndrome and visual acuity--the involvement of dynamic or acute sensory deprivation. AB - A 61-year-old patient suffered from Charles Bonnet syndrome (CBS) while his visual acuity declined, whereas CBS subsided after he became blind. These findings suggest that reduction of visual acuity (dynamic or acute impairment) has a greater impact on the onset of CBS than low visual acuity (static or chronic impairment) per se in some patients. They may also explain why patients with low visual acuity do not always suffer from CBS. Although further studies are required, the present case highlights the importance of the differentiation between lowering and low visual acuity in the etiology of CBS. PMID- 15538607 TI - The prevalence of tardive dyskinesia after a nine month naturalistic randomized trial comparing olanzapine with conventional treatment for schizophrenia and related disorders. AB - AIMS OF THE STUDY: To assess the impact of olanzapine versus conventional neuroleptic therapy among subjects with schizophrenia on ratings of tardive dyskinesia (TD). METHOD: The naturalistic study was conducted in three psychiatric hospitals in Brazil. Patients had a diagnosis of schizophrenia and related disorders (DSMIV) and with a BPRS score>24. Patients were evaluated by means of the PANSS scale for symptomatology (Kay et al. 1986), the Clinical Global Impression, The UKU side effect rating scale (Lingjaerde et al. 1987), and the Tardive Dyskinesia AIMS scale (Guy et al. 1976). Patients were seen by the treating physician routinely while hospitalized and then monthly on an out patient basis. All scale assessments were repeated after 9 months of discharge. RESULT: The sample was comprised of 190 patients (99 in the olanzapine and 91 in the standard treatment), with a completion rate of 88.2% for olanzapine and 84.9% for the conventional treatment (p=0.385, n. s.). The mean change from baseline in the PANSS total score favored olanzapine regarding negative symptoms (2.3, 95% C. I. 0.6-4.1, p<0.001); and general psychopathology (4.0, 95% C.I. 0.8-7.2, p<0.02) factors. TD was defined by applying Morgenstern & Glazer (1993) and Schooler & Kane (1982) criteria, on the basis of the AIMS scale. Both results favored olanzapine at the end of the follow-up (Morgenstern & Glazer: 25.6% versus 56.3%; Schooler & Kane: 16.3% versus 45.2%). At the end of the follow-up, by using the overall rating of the AIMS scale, the presence of TD was 2.3% for olanzapine (2/87), and 16.7% (12/72) for the conventional treatment. CONCLUSIONS: The results of this open label naturalistic trial showed that olanzapine had an impact on negative symptoms, decreased general psychopathology and reduced the risk of tardive dyskinesia. PMID- 15538608 TI - HPA axis activation determined by the CRH challenge test in patients with few versus multiple episodes of treatment-refractory depression. AB - OBJECTIVE: In clinical guidelines, risk factors for a malignant illness course include 3 or more lifetime episodes of depression. Our aim was to investigate the activation of the hypothalamic-pituitary-adrenal hormonal axis in treatment refractory affective disorder in pauciepisodic (one or two episodes) versus multiepisodic (three or more episodes) patients. METHODS: We evaluated the HPA axis in 37 patients with treatment-refractory affective disorder and in 27 healthy volunteers by measuring adrenocorticotropin hormone (ACTH) and cortisol responses following administration of corticotropin-releasing hormone (CRH). In retrospective life charts was recorded every previous illness episode for each patient. RESULTS: Seven of the patients were pauciepisodic and 30 were multiepisodic. The pauciepisodic patients had significantly larger peak and total ACTH responses to CRH compared to the multiepisodic patients as well as to the control group. Multiepisodic patients showed no difference compared to controls in ACTH secretion pre- and post-CRH. Cortisol secretion was the same in all three groups. CONCLUSIONS: The pituitary adrenocortical responses were stronger in pauciepisodic patients than in multiepisodic patients and in volunteers. This cross-sectional study suggests that the HPA axis, in refractory multiepisodic affective disorders, might weaken its original activity as the illness recurs with more episodes. PMID- 15538609 TI - Backstabbing--a report of an unusual case. AB - A case of a 25-year-old male with a deep knife stab injury in the back is reported. The stab wound penetrated the left thorax and the left lung was injured, a thoracotomy was performed and the patient survived. The injured man could not remember what had happened, his bag was missing and the incident was therefore considered to be due to a robbery. Further results of the police investigations and the forensic pathology inspection revealed an extremely unusual accident constellation. PMID- 15538610 TI - Quantification of secondary dentine formation from orthopantomograms--a contribution to forensic age estimation methods in adults. AB - The accuracy of most age determination methods in adults is unsatisfactory and therefore more precise methods are required. Teeth provide several useful points about an individuals age. With advancing age secondary dentine is deposited along the wall of the dental pulp chamber leading to a reduction in the size of the pulp cavity. These age-related changes can be determined and measured from dental radiographs. The aim of this study was to explore if measurements of the size of the pulp cavity performed on orthopantomograms can be used for individual age estimation. Measurements were made digitally for 6 types of teeth from orthopantomograms of 168 individuals aged between 14 and 81 years old. To compensate for errors due to the technique of radiography-for instance differences in angulation-ratios of distances were calculated: pulp/root length, pulp/tooth length and pulp/root width at three different root levels. The width ratios of the pulp cavity showed significant correlation to the chronological age and the coefficient of determination (r(2)) was highest in the upper lateral incisors (r(2)=0.913) when an exponential or a logistic regression model was constructed. At the same distance with a linear regression model the coefficient of determination (r(2)) reached 0.839. PMID- 15538611 TI - Validation of common classification systems for assessing the mineralization of third molars. AB - One major criterion for dental age estimation is the evaluation of third molar mineralization. There are various methods for evaluating tooth mineralization based on classification by stages. The aim of the present work is to assess the validity of the common classification systems. To this end, we analyzed 420 conventional orthopantomograms of German females aged 12-25 years old. The mineralization status of tooth 38 was determined using the stages defined by Gleiser and Hunt, Demirjian et al., Gustafson and Koch, Harris and Nortje and Kullman et al., respectively. Of the methods tested, the most accurate results were obtained with Demirjian et al.'s classification system, which performed best not only for observer agreement but also for the correlation between estimated and true age. It is argued that this is due to the fact that Demirjian et al.'s classification is based on a sufficient number of stages which are defined independently of speculative estimations of length. This leads to the conclusion that the method devised by Demirjian et al. should be used for evaluating the mineralization of third molars for purposes of forensic age determination. PMID- 15538612 TI - Nuclear telomerase is less accessible to antibody probing than known nuclear antigens: retrieval with new immunostaining buffer. AB - Telomerase is an important tumor marker but few antibodies to the enzyme have been described or used without difficulty in histochemical detection. Here we report specific detection of the enzyme in cell and tissue preparations using a new monoclonal antibody (mAb 476) and a new antigen-retrieval buffer (Enhancing buffer). When used to detect telomerase under normal immunostaining conditions in HL-60 cells or tissue sections of hepatocellular carcinoma or metastatic choriocarcinoma, unexpectedly, the antibody stained the cytoplasm rather than the nucleus. Nuclear staining, however, was revealed using the Enhancing buffer. Since other nuclear antigens in the HL-60 cell could be stained both ordinarily and in the Enhancing buffer, nuclear telomerase appears to be shrouded by the nuclear matrix or blocked by accessory proteins. The cytoplasmic activity seen in normal buffer but absent largely from the Enhancing buffer may be an artifact or the nascent, "naked" enzyme. With a known cytoplasmic antigen (proteinase-3) chosen arbitrarily for comparison, the antigenicity was found enhanced, instead, by the Enhancing buffer. The mode of action of the Enhancing buffer differs from that of microwave irradiation or the signal amplification (CSA) used by some investigators. The latter was found to enhance the cytoplasmic reactivity rather than the nuclear reactivity of mAb 476. PMID- 15538613 TI - Expression of TRPC homologs in endothelial cells and smooth muscle layers of human arteries. AB - TRPC channels are a group of Ca(2+)-permeable nonselective cation channels that mediate store-operated and/or agonist-stimulated Ca(2+) influx in a variety of cell types. In this study, we extensively examined the expression patterns of TRPC homologs in human vascular tissues. RT-PCR amplified cDNA fragments of TRPC1 (505 bp), TRPC3 (372 bp), TRPC4 (499 bp), TRPC5 (325 bp), TRPC6 (509 bp), and TRPC7 (187 bp) from RNA isolated from cultured human coronary artery endothelial cells. In situ hybridization yielded strong labeling of TRPC1,3-6 in the endothelial and smooth muscle cells of human coronary and cerebral arteries. TRPC7 labeling was exclusively found in endothelial cells but not in smooth muscle cells. Results from immunohistochemical staining were consistent with those from in situ hybridization. Similar expression patterns of TRPC homologs were also observed in arterioles and vaso vasora. In conclusion, our study indicates that TRPC homologs are widely expressed in human vessels of all calibers, including medium-sized coronary arteries and cerebral arteries, smaller sized resistance arteries, and vaso vasora. These results suggest a ubiquitous role of TRPC homologs in regulating blood supply to different regions and in controlling arterial blood pressure. PMID- 15538614 TI - The Epping Jaundice outbreak: mortality after 38 years of follow-up. AB - OBJECTIVE: To investigate mortality in individuals exposed to 4,4' methylenedianiline (MDA). METHODS: The mortality of 84 individuals, accidentally poisoned with MDA during the "Epping Jaundice" outbreak of 1965, was compared with expected values based on national rates defined by age, period and sex, for the period 1965-2002. In addition, cancer registration data were analysed for the period 1971-2002. RESULTS: The vital status of 83% of the group was established, with 37 deaths occurring before the end of follow-up. Mortality from all causes was close to expectation among females (Obs, 25; Exp, 30.3; SMR, 82), and below expectation among males (Obs, 12; Exp, 26.7; SMR, 45). There were no observed deaths from cancer of the liver or from nonmalignant liver disease. CONCLUSIONS: This study has found no evidence to suggest that the ingestion of MDA had adversely affected mortality. PMID- 15538615 TI - Respiratory and general health impairments of ragpickers in India: a study in Delhi. AB - OBJECTIVE: This study attempted to assess the respiratory and general health of ragpickers who rummage through the garbage dumps and landfill sites in India to collect and sell recyclable materials for a living. METHODS: 98 ragpickers and 60 controls from Delhi, matched for age, sex, and socioeconomic conditions, were examined. Health data were obtained from questionnaire survey, clinical examination, and laboratory investigations. Lung function was evaluated by spirometry. RESULTS: After controlling for smoking as a confounder, respiratory symptoms and lung function decrement were recorded in 94% and 52% of the ragpickers, respectively, compared with 56% and 34% of controls. The ragpickers showed a higher prevalence of low hemoglobin, high circulating eosinophil and monocyte counts, unhealthy gums, frequent diarrhea, and dermatitis, when compared with controls. Their sputum showed an abundance of alveolar macrophages, siderophages and inflammatory cells, and a very high frequency of squamous metaplasia and dysplasia of bronchial epithelial cells, suggesting inflammation and cellular changes in the airways. CONCLUSION: The ragpickers suffer from a multitude of health problems which seem related to their occupation. PMID- 15538616 TI - Levels of agreement of nerve conduction studies and symptoms in workers at risk of carpal tunnel syndrome. AB - OBJECTIVE: Information is required on levels of agreement between different sets of median nerve conduction studies (NCS) and symptoms of carpal tunnel syndrome (CTS) in at-risk subjects. METHODS: We performed a reanalysis of an extensive data set from a population of 114 "highly exposed" meat workers (without prior diagnosis of CTS), who were interviewed for the presence of current symptoms typical of CTS and systematically submitted to median NCS. RESULTS: Agreement between presence/absence of symptoms and NCS normal/abnormal findings ranged from 81% for wrist sensory latency (WSL) to 49% for motor conduction velocity wrist-to palm (MCV-WP) in the nondominant hands (from 60% to 50%, respectively, in the dominant hands). Mean values of WSL, sensory conduction velocity-WP (SCV-WP), wrist motor latency, MCV-WP, and the SCV-WP/SCV-elbow-to-wrist ratio all showed significant differences between symptomatic and asymptomatic workers. In nondominant hands, these parameters all reached significant P values at chi squared tests for kappa coefficients of agreement, the best kappa coefficients being 0.57 for WSL and 0.40 for SCV-WP. However, in the dominant hand only SCV-WP reached significance (kappa coefficient=0.20). CONCLUSIONS: Given the importance of the dominant hand in working populations, these data support use of SCV-WP (or WSL) as an informative NCS parameter for occupational studies on CTS. PMID- 15538617 TI - Monitoring of chromium and nickel in biological fluids of stainless steel welders using the flux-cored-wire (FCW) welding method. AB - OBJECTIVE: This study was undertaken to investigate the exposure to chromium (Cr) and nickel (Ni) in flux-cored wire (FCW) welders welding on stainless steel (SS). METHOD/DESIGN: Seven FCW welders were monitored for 3 days to 1 workweek, measuring Cr and Ni in air, blood, and urine. The welders were questioned about exposure to Cr and Ni during their whole working careers, with emphasis on the week of monitoring, about the use of personal protective equipment and their smoking habits. RESULTS: The air concentrations were mean 200 microg/m(3) (range 2.4-2,744) for total Cr, 11.3 microg/m(3) (<0.2-151.3) for Cr(VI), and 50.4 microg/m(3) (<2.0-416.7) for Ni during the workdays for the five welders who were monitored with air measurements. The levels of Cr and Ni in biological fluids varied between different workplaces. For Cr in whole blood, plasma, and erythrocytes, the mean levels after work were 1.25 (<0.4-8.3) and 1.68 (<0.2-8.0) and 0.9 (<0.4-7.2) microg/l, respectively. For Ni most of the measurements in whole blood and plasma were below the detection limits, the mean levels after work being 0.84 (<0.8-3.3) and 0.57 microg/l (<0.4-1.7), respectively. Mean levels for Cr and Ni in the urine after work were 3.96 (0.34-40.7) and 2.50 (0.56 5.0) microg/g creatinine, respectively. CONCLUSION: Correlations between the Cr(VI) levels measured in air and the levels of total Cr in the measured biological fluids were found. The results seem to support the view that monitoring of Cr in the urine may be versatile for indirect monitoring of the Cr(VI) air level in FCW welders. The results seem to suggest that external and internal exposure to Cr and Ni in FCW welders welding SS is low in general. PMID- 15538618 TI - Reliability and validity of Functional Capacity Evaluation methods: a systematic review with reference to Blankenship system, Ergos work simulator, Ergo-Kit and Isernhagen work system. AB - OBJECTIVES: Functional Capacity Evaluation methods (FCE) claim to measure the functional physical ability of a person to perform work-related tasks. The purpose of the present study was to systematically review the literature on the reliability and validity of four FCEs: the Blankenship system (BS), the ERGOS work simulator (EWS), the Ergo-Kit (EK) and the Isernhagen work system (IWS). METHODS: A systematic literature search was conducted in five databases (CINAHL, Medline, Embase, OSH-ROM and Picarta) using the following keywords and their synonyms: functional capacity evaluation, reliability and validity. The search strategy was performed for relevance in titles and abstracts, and the databases were limited to literature published between 1980 and April 2004. Two independent reviewers applied the inclusion criteria to select all relevant articles and evaluated the methodological quality of all included articles. RESULTS: The search resulted in 77 potential relevant references but only 12 papers were identified for inclusion and assessed for their methodological quality. The interrater reliability and predictive validity of the IWS were evaluated as good while the procedure used in the intrarater reliability (test-retest) studies was not rigorous enough to allow any conclusion. The concurrent validity of the EWS and EK was not demonstrated while no study was found on their reliability. No study was found on the reliability and validity of the BS. CONCLUSIONS: More rigorous studies are needed to demonstrate the reliability and the validity of FCE methods, especially the BS, EWS and EK. PMID- 15538619 TI - Coping and self-image in patients with visual display terminal-related skin symptoms and perceived hypersensitivity to electricity. AB - OBJECTIVES: The aim of the present study was to measure coping resources and self image in patients with visual display terminal (VDT)-related skin symptoms and hypersensitivity to electricity (HE). METHODS: From 1980 to 1998, 350 patients with electrical sensitivity were registered. The patients were subdivided into two groups: patients with skin symptoms evoked by VDTs, television screens, and fluorescent-light tubes and patients with so-called hypersensitivity to electricity with multiple symptoms evoked by exposure to different electrical environments. A questionnaire was sent to all patients and contained the coping resources inventory (CRI) and the structural analysis of social behaviour (SASB) in order for us to measure coping resources and self-image, respectively. The CRI and SASB scores were compared with those of control groups. Two hundred and fifty respondents (73%) returned the questionnaire, 200 (78.5% women) in the VDT group and 50 (62% women) in the HE group. RESULTS: The patient group rated high on the CRI spiritual/philosophical scale and high on the SASB spontaneous, positive and negative clusters but low on the controlled cluster. The female patients scored high on the CRI emotional scale. The VDT group rated lower than the controls on the SASB controlled cluster and higher on both the positive and negative cluster. The HE group scored higher than the control group on the SASB spontaneous and positive clusters. The women in the HE group scored higher on the CRI cognitive and CRI total scale than the VDT group and control group and higher on the CRI emotional scale than the controls. The women in the HE group rated higher than both the women in the VDT and control groups on the SASB spontaneous and positive clusters. CONCLUSIONS: The deviant self-image found in these patients, especially the female HE patients, support the view that VDT and HE symptoms can be stress related. In the clinic, a trustful alliance should be established with the patient in order for a more realistic view to be achieved of the capacity. PMID- 15538620 TI - Contact sensitizations in metalworkers with occupational dermatitis exposed to water-based metalworking fluids: results of the research project "FaSt". AB - BACKGROUND: The composition of water-based metalworking fluids (wb MWF) is complex, and various admixtures may be added before or during usage. Wb MWF may cause irritant as well as allergic contact dermatitis. While several current case reports point towards allergens particularly related to wb MWF, systematic studies have not been performed for several years. From 1999 to 2001, a study on contact allergies among patients with occupational dermatitis (OD) called "Fruhzeitige Erkennung allergener Stoffe bei beruflicher und nicht-beruflicher Exposition" (German acronym: FaSt) was conducted by the Information Network of Departments of Dermatology (IVDK), funded by the employers' liability insurances in Germany (HVBG). OBJECTIVE: The objective of FaSt was to detect sensitization patterns related to particular occupational exposures. METHODS: Anamnestic and clinical data were gathered using a standardised questionnaire. Patch test results were recorded by computer within the IVDK routine procedure. In addition to descriptive statistical analyses, logistic regression analysis was performed to control the effect of potential confounders. RESULTS: Among the 1842 OD patients in the FaSt study, there were 160 metalworkers exposed to wb MWF, whose data is presented in this paper. A specific allergen pattern of these patients can be described: most frequently, sensitizations to monoethanolamine (MEA), colophony/abietic acid, and fragrance mix were observed. Additionally, cobalt, formaldehyde, formaldehyde releasers and other biocides are important allergens in these patients. CONCLUSIONS: Preventive measures and aimed in-depth research may be based on these results. The special MWF test series have to be kept up to date based on exposure information from the MWF industry and on continuous surveillance of the target group. PMID- 15538621 TI - Recurrence risks in randomized trials of laparoscopic versus open inguinal hernia repair: to pool or not to pool (this is not the question). AB - BACKGROUND AND AIMS: Meta-analyses suggest that, with regard to relative risks of hernia recurrence, there is a net benefit in the use of synthetic mesh rather than the surgical approach. Considering important arguments raised by both conventional and laparoscopic surgeons, we set out to explore other sources of variability across individual studies. METHODS: We abstracted all publications of randomized trials of laparoscopic versus open inguinal hernia repair included in the EU Hernia Trialists meta-analyses. We applied meta-regression to identify variables that are likely to alter the relative risk of hernia recurrence with either route. RESULTS: We included 41 randomized trials (7,446 patients), two of which were identified by a systematic literature search. There was significant statistical heterogeneity across studies (chi(2) test P=0.029). Meta-regression was limited because of scarce information provided in the original papers, and small sample sizes. Results varied internationally, with trials from the UK, southern Europe and Australia favouring open hernioplasty (analysis of variance, P=0.0047). Large numbers of surgeons contributing to the open hernioplasty group predicted better results with endoscopic hernia repair [risk ratio (RR)] 0.99 with any additional surgeon, 95% confidence interval (CI) 0.98-1.00, P=0.005]. Non-significant trends were observed towards reduced recurrence risks with increasing mesh sizes. Training procedures performed before patient enrollment slightly reduced the relative risk of recurrence with endoscopic hernioplasty. CONCLUSION: Because of the diversity in the size of effect, it is doubtful whether data from the available hernia trials should be compiled into a single summary measure. Efficacy estimates in hernia surgery are susceptible to technical issues, which need further scientific appraisal on a larger scale. PMID- 15538622 TI - Isolation and functional characterization of the Ca-DREBLP1 gene encoding a dehydration-responsive element binding-factor-like protein 1 in hot pepper (Capsicum annuum L. cv. Pukang). AB - Through the use of subtractive hybridization analysis, we have identified 14 partial cDNA clones (pCa-DSRs) that are rapidly induced by dehydration in hot pepper (Capsicum annuum L.) roots. The predicted proteins encoded by Ca-DSRs are putatively involved in processes as diverse as primary and secondary metabolism, protein degradation, and stress responses, indicating the complexity of cellular responses to water deficit in hot pepper roots. Particularly, we investigated the detailed structural properties and expression profiles of Ca-DSR2 (Ca-DREBLP1: dehydration-responsive element binding-factor-like protein 1) encoding a protein that contains a single ERF/AP2 DNA-binding domain. Based on the conserved 14th valine and 19th glutamic acid residues in the ERF/AP2 domain, a basic amino acid stretch (PKKPAGRKKFR) near its N-terminal region, and DSAW signature sequence at the end of its ERF/AP2 domain, Ca-DREBLP1 was classified as a member of a DREB1 type subfamily. Gel retardation assays revealed that Ca-DREBLP1 was able to form a specific complex with the DRE/CRT motif, but not with the GCC box. When fused to the GAL4 DNA-binding domain, the Ca-DREBLP1(190-215) mutant could effectively function as a trans-activator in yeast. This suggests that the extreme C-terminal region plays an essential role in transcription activation. In hot pepper plants, Ca-DREBLP1 was rapidly induced by dehydration, high salinity and, to a lesser extent, mechanical wounding, but not by cold stress. Thus, although the structural features of Ca-DREBLP1 resemble those of the DREB1-type proteins of Arabidopsis thaliana and rice plants, its induction patterns are reminiscent of the DREB2-type proteins, indicating that Ca-DREBLP1 is a novel class DREB subfamily in hot pepper. PMID- 15538625 TI - Lipomatosis of the penis and perineum in a 6-year-old boy. PMID- 15538626 TI - Complexity in the treatment of pulmonary large cell neuroendocrine carcinoma. AB - PURPOSE: According to the World Health Organization (WHO) classification of pulmonary large cell neuroendocrine carcinoma (LCNEC), one of the neuroendocrine tumors of the lung, is considered as a variant of non-small cell lung carcinoma. The objective of this study was to investigate the treatment strategy for LCNEC. METHODS: We retrospectively reviewed the clinical information of 12 patients with LCNEC. RESULTS: Three patients with stage I disease underwent curative resection but all relapsed within 20 months. One with stage IIA disease underwent non curative resection received adjuvant chemoradiotherapy (cisplatin plus etoposide) and is well with no evidence of recurrence. Two with stage IIIB disease received concurrent chemoradiotherapy. Both achieved partial response (PR) but relapsed within 2 months. One elderly patient with stage IIIA disease received vinorelbine alone and did not respond. Of five patients with stage IV disease, three received platinum-based chemotherapy but no patient achieved PR. Of five patients with gefitinib as salvage therapy, one achieved PR. CONCLUSIONS: The prognosis of LCNEC is poor. To improve the outcome, we must evaluate the effectiveness of adjuvant or neoadjuvant therapy in patients with resectable disease. In addition, the evaluation of systemic and multimodality treatment strategies similar as in small cell lung cancer is worthy of consideration. PMID- 15538627 TI - 5-fluorouracil and hydroxyurea enhance adenovirus-mediated transgene expression in colon and hepatocellular carcinoma cells. AB - PURPOSE: To investigate the efficient transduction of tumor cells which remains a major limitation of cancer gene therapy. METHODS: In this study, we tested whether treatment with antimetabolic drugs 5-FU and hydroxyurea (HU) could improve adenovirus-mediated gene expression in tumor cells. RESULTS: We found that 5-FU and HU treatment significantly increased beta-gal activity in adenovirus (Ad.CMVBG)-infected human colon carcinoma (LoVo) and hepatocellular carcinoma (SMMC7721) cells in a dose- and time-dependent manner. These increases were maximized at 5.01+/-0.42-fold and 3.32+/-0.32-fold for 5-FU (50 microM), and at 6.60+/-0.50-fold and 4.82+/-0.43-fold for HU (5 mM) treatment, respectively, after 48 h infection. Transient increases in viral uptake, determined by real time PCR for viral DNA content and by confocal microscopy for viral particles, were observed in 5-FU or HU-treated cells that partially contribute to the overall increases of beta-gal expression. Moreover, mRNA levels for the beta-gal gene in infected cells were significantly increased in both LoVo and SMMC7721 cells by 5-FU and HU treatment in contrast to the inhibition of viral DNA replication and the unchanged mRNA levels for alpha-actin gene. The induction appeared to be the result of enhanced transcription since beta-gal mRNA half-life was not affected by drug treatment. However, similar induction was not detected in CMV-beta-gal-expressing stable cells, suggesting that an adenovirus-associated mechanism might be involved in this induction. CONCLUSIONS: Our findings suggest that it may be possible to improve tumor cell transduction by adenovirus using chemotherapy. PMID- 15538628 TI - Cadmium and lead concentrations in Gallegoides arfaai (Cestoda: Anoplocephalidae) and Apodemus sylvaticus (Rodentia: Muridae) from Spain. AB - Information on parasites of vertebrates living in terrestrial ecosystems as sentinels for heavy metal environmental pollution is scarce. The aim of the present study was to assess the concentration of cadmium and lead using the model Apodemus sylvaticus/Gallegoides arfaai in order to test the potential suitability of G. arfaai as a sentinel organism for lead and cadmium under natural field conditions. Samples of 15 A. sylvaticus as well as whole specimens of G. arfaai were analysed for both elements by inductively coupled plasma mass spectrometry. The level of cadmium in G. arfaai was always much lower than in the tissues of A. sylvaticus. Contrarily, values for lead in G. arfaai were found to be 6, 20 and 24-fold higher than in the kidney, liver and muscle of A. sylvaticus. We propose the model A. sylvaticus/G. arfaai as a promising bioindication system to evaluate environmental lead exposure in terrestrial habitats, especially for non-urban areas. PMID- 15538629 TI - Novel beta-carotene ketolases from non-photosynthetic bacteria for canthaxanthin synthesis. AB - We reported previously that the Rhodococcus erythropolis strain AN12 synthesizes the monocyclic carotenoids 4-keto gamma-carotene and gamma-carotene. We also identified a novel lycopene beta-monocyclase in this strain. Here we report the identification of the rest of the carotenoid synthesis genes in AN12. Two of these showed apparent homology to putative phytoene dehydrogenases. Analysis of Rhodococcus knockout mutants suggested that one of them ( crtI) encodes a phytoene dehydrogenase, whereas the other ( crtO) encodes a beta-carotene ketolase. Expression of the beta-carotene ketolase gene in an Escherichia coli strain which accumulates beta-carotene resulted in the production of canthaxanthin. In vitro assays using a crude extract of the E. coli strain expressing the crtO gene confirmed its ketolase activity. A crtO homologue (DR0093) from Deinococcus radiodurans R1 was also shown to encode a beta-carotene ketolase, despite its sequence homology to phytoene dehydrogenases. The Rhodococcus and Deinococcus CrtO ketolases both catalyze the symmetric addition of two keto groups to beta-carotene to produce canthaxanthin. Even though this activity is similar to the CrtW-type of ketolase activity, the CrtO ketolases show no significant sequence homology to CrtW-type ketolases. The presence of six conserved regions may be a signature for the CrtO-type of beta-carotene ketolases. PMID- 15538630 TI - Novel and recurrent mutations in the laminin-5 genes causing lethal junctional epidermolysis bullosa: molecular basis and clinical course of Herlitz disease. AB - Herlitz disease (H-JEB), the lethal form of junctional epidermolysis bullosa, is a rare genodermatosis presenting from birth with widespread erosions and blistering of skin and mucosae because of tissue cleavage within the epidermal basement membrane. Mutations in any of the three genes encoding the alpha3, beta3 and gamma2 chains of laminin-5 underlie this recessively inherited disorder. Here, we report the molecular basis and clinical course of H-JEB in 12 patients. Two novel nonsense mutations in the gene LAMA3 (E281X and K1299X) and a novel frame-shift mutation in the gene LAMB3 (1628insG) leading to a premature termination codon were identified by DNA sequencing and confirmed by restriction fragment length polymorphism analysis. In the four patients affected, neither the resulting truncated polypeptide chains nor assembled laminin-5 protein were detectable by immunofluorescence. Three patients were found to be heterozygous for the known hotspot mutation R635X and the recurrent mutations Q373X or 29insC in the gene LAMB3, whereas five others were homozygous for R635X. Significant variations in the disease progression and survival times between 1 and 30 months in this group of H-JEB patients emphasised the impact of modifying factors and the importance of immunostaining or mRNA assessment as parallel diagnostic methods. Interestingly, the only patients who survived for longer than 6 months were four females carrying the mutation R635X homozygously. In one of them, the clinical course may have been improved by treatment with artificial skin equivalents. These data may stimulate further investigation of genotype-phenotype correlations and facilitate mutation analysis and genetic counselling of affected families. PMID- 15538631 TI - Characterization of six novel mutations in the CYBB gene leading to different sub types of X-linked chronic granulomatous disease. AB - Chronic granulomatous disease is an inherited disorder in which phagocytes lack a functional NADPH oxidase and so cannot generate superoxide anions (O(2) (-)). The most common form is caused by mutations in CYBB encoding gp91 phox, the heavy chain of flavocytochrome b(558) (XCGD). We investigated 11 male patients and their families suspected of suffering from X-linked CGD. These XCGD patients were classified as having different variants (X91(0), X91(-) or X91(+)) according to their cytochrome b(558) expression and NADPH oxidase activity. Nine patients had X91(0) CGD, one had X91(-) CGD and one had X91(+) CGD. Six mutations in CYBB were novel. Of the four new X91(0) CGD cases, three were point mutations: G65A in exon 2, G387T in exon 5 and G970T in exon 9, leading to premature stop codons at positions Try18, Try125 and Glu320, respectively, in gp91 phox. One case of X91(0) CGD originated from a new 1005G deletion detected in exon 9. Surprisingly, four nonsense mutations in exon 5 led to the generation of two mRNAs, one with a normal size containing the mutation and the other in which exon 5 had been spliced. A novel X91(-) CGD case with low gp91 phox expression was diagnosed. It was caused by an 11-bp deletion in the linking region between exon 12 and intron 12, activating a new cryptic site. Finally, a new X91(+) CGD case was detected, characterized by a missense mutation Leu505Arg in the potential NADPH-binding site of gp91 phox. No clear correlation between the severity of the clinical symptoms and the sub-type of XCGD could be established. PMID- 15538632 TI - A new locus for nonsyndromic deafness DFNB49 maps to chromosome 5q12.3-q14.1. AB - Cosegregation of markers on chromosome 5q12.3-q14.1 with profound congenital deafness in two Pakistani families (PKDF041 and PKDF141) defines a new recessive deafness locus, DFNB49. A maximum two-point lod score of 4.44 and 5.94 at recombination fraction theta=0 was obtained for markers D5S2055 and D5S424 in families PKDF041 and PKDF141, respectively. Haplotype analysis revealed an 11 cM linkage region flanked by markers D5S647 (74.07 cM) and D5S1501 (85.25 cM). Candidate deafness genes in this region include SLC30A5, OCLN, GTF2H2, and BTF3, encoding solute carrier family 30 (zinc transporter) member 5, occludin, RNA polymerase II transcription initiation factor, and basic transcription factor 3, respectively. Sequence analysis of the coding exons of SLC30A5 in DNA samples from two affected individuals of families PKDF041 and PKDF141 revealed no mutation. The mapping of DFNB49 further confirms the heterogeneity underlying autosomal recessive forms of nonsyndromic deafness. PMID- 15538633 TI - Photosynthetic responses to vapour pressure deficit in temperate and tropical evergreen rainforest trees of Australia. AB - Rainforests occur in high precipitation areas of eastern Australia, along a gradient in seasonality of precipitation, ranging from a summer dry season in the temperate south to a winter dry season in the tropical north. The response of net photosynthesis to increasing vapour pressure deficit (VPD) was measured in a range of Australian rainforest trees from different latitudes to investigate possible differences in their response to atmospheric drought. Plants were grown in glasshouses under ambient or low VPD to determine the effect of growth VPD on the photosynthetic response. Temperate species, which experience low summer precipitation, were found to maintain maximum net photosynthesis over the measurement range of VPD (0.5-1.9 kPa). In contrast, the tropical species from climates with high summer precipitation showed large reductions in net photosynthesis with increasing VPD. Temperate species showed higher intrinsic water-use efficiencies under low VPD than the tropical species, whereas their efficiencies were similar under high VPD. Growing plants under a low VPD had little effect on either the photosynthetic response to VPD or the intrinsic water use efficiency of the species. These different responses of gas exchange to VPD shown by the tropical and temperate rainforest species may reflect different strategies to maximise productivity in their respective climates. PMID- 15538634 TI - Effects of plant diversity, community composition and environmental parameters on productivity in montane European grasslands. AB - In the past years, a number of studies have used experimental plant communities to test if biodiversity influences ecosystem functioning such as productivity. It has been argued, however, that the results achieved in experimental studies may have little predictive value for species loss in natural ecosystems. Studies in natural ecosystems have been equivocal, mainly because in natural ecosystems differences in diversity are often confounded with differences in land use history or abiotic parameters. In this study, we investigated the effect of plant diversity on ecosystem functioning in semi-natural grasslands. In an area of 10x20 km, we selected 78 sites and tested the effects of various measures of diversity and plant community composition on productivity. We separated the effects of plant diversity on ecosystem functioning from potentially confounding effects of community composition, management or environmental parameters, using multivariate statistical analyses. In the investigated grasslands, simple measures of biodiversity were insignificant predictors of productivity. However, plant community composition explained productivity very well (R2=0.31) and was a better predictor than environmental variables (soil and site characteristics) or management regime. Thus, complex measures such as community composition and structure are important drivers for ecosystem functions in semi-natural grasslands. Furthermore, our data show that it is difficult to extrapolate results from experimental studies to semi-natural ecosystems, although there is a need to investigate natural ecosystems to fully understand the relationship of biodiversity and ecosystem functioning. PMID- 15538635 TI - Erosion and landscape development affect plant nutrient status in the Hawaiian Islands. AB - We quantified variation in plant nutrient concentrations and provenance along catenas in landscapes of three different ages (0.15, 1.4, and 4.1 ma) in the Hawaiian Islands. Strontium (Sr) isotopes demonstrate that erosion provides a renewed source of rock-derived nutrients to slopes in landscapes of all ages, in some cases reversing a million years of ecosystem development in a distance of 100 m. However the effects of this input vary with landscape age. Plants on uneroded surfaces in a 0.15-ma landscape derive approximately 20% of their Sr from local bedrock (foliar 87Sr/86Sr approximately 0.7085), while on adjacent slopes this increases to approximately 80% (foliar 87Sr/86Sr approximately 0.7045). Despite this shift in provenance, foliar N and P do not vary systematically with slope position. Conversely, eroded slopes in a 4.1-ma landscape show smaller increases in rock-derived cations relative to stable uplands (foliar 87Sr/86Sr approximately 0.7075 vs 0.7090), but have >50% higher foliar N and P. These results demonstrate both that erosion can greatly increase nutrient availability in older landscapes, and that the ecological effects of erosion vary with landscape age. In addition, there can be as much biogeochemical variation on fine spatial scales in eroding landscapes as there is across millions of years of ecosystem development on stable surfaces. PMID- 15538636 TI - Competitive equivalence maintains persistent inter-clonal boundaries. AB - Clear boundaries often separate adjacent conspecific competitors. These boundaries may reflect bordering animal territories or regions of inter-organism contact in mobile and non-mobile organisms, respectively. Sessile, clonal organisms often form persistent inter-clonal boundaries despite great variation in competitive ability among genotypes within a population. I show that neighboring clones in the sea anemone Anthopleura elegantissima and three species of the marine hydroid genus Hydractinia are more evenly matched in terms of competitive ability than expected by chance. Hypotheses of genetic relatedness or similar environmental regime shared by neighboring clones are inconsistent with the observed similarities between adjacent competitors in one or both taxa. Instead, inter-clonal borders evidently persist as standoffs between evenly matched competitors. Large differences in competitive ability between bordering clones were rarely observed, suggesting that dominant clones quickly displace or eliminate others in competitive mismatches. This ecological parallel between taxa (i.e., competitive equivalence) exists despite several fundamental differences (e.g., geographical distribution, habitat, body size, longevity), suggesting that competitive equivalence may be a widespread determinant of boundary persistence between adjacent competitors. PMID- 15538637 TI - The effect of climate on the phenology, acorn crop and radial increment of pedunculate oak (Quercus robur) in the middle Volga region, Tatarstan, Russia. AB - Our data, collected in the extreme east of Europe, show that a significant biological effect of climate change has been experienced even in territories where temperature increase has been the lowest. This study documents the climatic response of pedunculate oak (Quercus robur) growing near its north-eastern limits in Europe. It demonstrates the potential of oak trees in old-growth forest to act as proxy climate indicators. Many factors may influence the temporal stability of the growth-climate, acorn crop-climate and first leafing-climate relationships. Climate data, climatic fluctuations, reproduction, genetics and tree-age may relate to this instability. Our results stress that an increase in climate variability or climatic warming resulting from warmer winters or summers could affect the oak population in eastern Europe in a similar way to that in western Europe. These findings, from remnants of oak forest in the middle Volga region of Russia, allow a further understanding of how species could be affected by future climates. PMID- 15538638 TI - Effectiveness and safety of oral extended-release oxymorphone for the treatment of cancer pain: a pilot study. AB - GOALS OF WORK: Inadequate analgesia and/or unmanageable adverse events frequently result in the need to rotate patients with cancer pain to a different opioid. The availability of a novel oral extended-release (ER) formulation of oxymorphone provides clinicians with another treatment option. In this study, we assessed the analgesic effectiveness and safety of the new oral ER formulation of oxymorphone following treatment with controlled-release (CR) morphine sulfate or oxycodone. PATIENTS AND METHODS: Adults with moderate to severe cancer pain were stabilized for > or =3 days on morphine CR or oxycodone CR, and then treated for 7 days at their stabilized dose. Drug selection was based upon patients' previous use or investigator preference. Patients were then crossed over for 7 days of treatment at an estimated equianalgesic dosage of oxymorphone ER. Pain was assessed using a visual analog scale, and adverse events were recorded. MAIN RESULTS: A total of 86 patients entered open-label treatment. Of 34 patients assigned to morphine CR and 52 assigned to oxycodone CR, 21 (61.8%) and 42 (80.8%) completed stabilization and began treatment with oxymorphone ER, respectively; 59 of 63 (93.7%) completed treatment with oxymorphone. There were no significant differences in daily pain intensity scores between oxymorphone ER and comparators (paired t -test). Rescue medication use, expressed as the percent of the daily dose of scheduled opioid, was greater during morphine CR treatment than after crossover to oxymorphone ER (25.2% vs 13.3%; P <0.05, Wilcoxon's test). The tolerability/safety profiles (e.g., nausea, drowsiness, somnolence) were similar for all opioids. CONCLUSIONS: Cancer patients stabilized on morphine CR or oxycodone CR were safely and rapidly converted to a lower milligram dose of oxymorphone ER that provided adequate pain relief with comparable tolerability. These results justify additional trials with oxymorphone ER. PMID- 15538639 TI - Catheter tip position as a risk factor for thrombosis associated with the use of subcutaneous infusion ports. AB - AIMS: The use of subcutaneous infusion ports has become standard practice to provide a long-term venous access in oncological patients. The aim of this retrospective study was to assess the different complications of infusion ports in our population and to identify predisposing factors. PATIENTS AND METHODS: We reviewed the medical records of 437 patients who were followed at the Oncology/Haematology Department of our hospital. Of these patients, there were 370 (84.4%) with solid tumours and 58 (13.2%) with haematological disease. The position of the catheter tip was evaluated by reviewing the available chest radiographs or phlebographies. MAIN RESULTS: Analysis of the records showed that 346 patients (79.17%) had no complications. The most common complications after implantation were thrombosis (8.46%), catheter dysfunction (4.8%) and infections (4.4%). Univariate and multivariate analysis showed that catheter tip positioning was the most important predisposing risk factor for thrombosis. Catheter tips positioned in the brachiocephalic vein or in the cranial part of the superior vena cava were associated with a high risk of thrombosis. Other significant risk factors were gender and initial diagnosis. Female patients and patients with lung cancer also had an elevated risk of developing a thrombosis. CONCLUSIONS: Compared to other reports, we noted a higher rate of thrombosis and port dysfunction. Since catheter tip position was a predisposing factor for developing a thrombosis, correct catheter position has to be ensured during placement. Prophylactic antithrombotic treatment might be beneficial in the event of failure to position the catheter correctly. PMID- 15538640 TI - Rankings and symptom assessments of side effects from chemotherapy: insights from experienced patients with ovarian cancer. AB - GOALS OF WORK: Although many patients with ovarian cancer achieve favorable responses to primary chemotherapy, the majority of women will experience recurrence of their cancer. Selection of second- or third-line chemotherapy ultimately depends on patient preferences for different side effects. To better understand this process, we evaluated preferences and symptom distress in patients with ovarian cancer. PATIENTS AND METHODS: A total of 70 women with ovarian cancer who had previously received at least three cycles of platinum based chemotherapy and currently undergoing chemotherapy for newly diagnosed or recurrent disease were interviewed in an outpatient chemotherapy clinic. The patients were asked to rank order 27 health states using a modified visual analog scale and to complete the Memorial Symptom Assessment Scale (MSAS). MAIN RESULTS: Most favorable health states included perfect health, clinical remission and complete control of chemotherapy-induced nausea and vomiting (CINV). Least favorable health states included more severe CINV health states and death. Patients on first-line chemotherapy had less symptom distress, and rated sexual dysfunction, fatigue and memory loss more favorably than patients on second- or third-line chemotherapy (P<0.05). Married patients generally had less symptom distress compared to patients who were not married, but married patients indicated more distress with sexual dysfunction (P=0.04). Married patients rated alopecia less favorably than unmarried patients (P=0.03), but married patients viewed certain CINV health states more favorably (P=0.02-0.04). CONCLUSIONS: CINV remains one of the most dreaded side effects of chemotherapy. Separate preference profiles exist for patients with newly diagnosed and recurrent disease, as well as for married versus unmarried patients. While MSAS scores and VAS rankings showed consistency across some health states, this was not true for CINV, suggesting that current symptom status may only influence patient preferences for selected side effects. PMID- 15538641 TI - Review of telephone follow-up experience at the Rapid Response Radiotherapy Program. AB - PURPOSE: To review the feasibility of telephone follow-up following a 3-year experience from 1999 to 2001 at the Rapid Response Radiotherapy Program as a prospective radiotherapy outcome assessment of symptom palliation. METHODS AND MATERIALS: Patients referred for palliative radiotherapy for symptom control from 1999 to 2001 were asked to rate symptom distress using the Edmonton Symptom Assessment System (ESAS) at initial consultation. Patient demographics and analgesic consumption were recorded. Telephone follow-up interviews were conducted with all patients who received radiotherapy during weeks 1, 2, 4, 8, and 12 after radiotherapy using the ESAS and an analgesic diary. Clinic follow-up visits were conducted only if necessary. RESULTS: Between January 1999 and December 2001, 830 patients received palliative radiotherapy. Of these patients, 260 (31.3%) died during the 12-week follow-up period. The percentage of surviving patients responding to the telephone interview ranged from 48.2% (week 12) to 56.6% (week 4) during the 12-week study. CONCLUSION: Telephone follow-up is a feasible tool for prospective outcome assessment of symptom palliation in metastatic cancer patients. Telephone follow-up is recommended for outcome assessment of symptom relief when it can achieve the same purpose as clinic follow-up. Data collection remains a challenge in the palliative patient population. Further studies are required to determine how well other methods of maximizing data collection, such as through the use of caregivers, can provide reliable and accurate information. PMID- 15538642 TI - Antiemetics in children receiving chemotherapy. AB - Only a few studies have been carried out in children on the prevention of chemotherapy-induced acute emesis. 5-HT3 antagonists have been shown to be more efficacious and less toxic than metoclopramide, phenothiazines and cannabinoids. The optimal dose and scheduling of the 5-HT3 antagonists has not been identified. Combinations of a 5-HT3 antagonist and dexamethasone show increased efficacy with respect to 5-HT3 antagonists alone. All pediatric patients receiving chemotherapy of high or moderate emetogenic potential should receive a combination of a 5-HT3 antagonist and dexamethasone to prevent acute emesis. No studies have specifically evaluated antiemetic drugs in the prevention of chemotherapy-induced delayed and anticipatory emesis in children. PMID- 15538643 TI - Mass spectrometrical analysis of phosphoprotein enriched in astrocytes of 15 kDa in mouse hippocampi. AB - Phosphoprotein enriched in astrocytes of 15 kDa (PEA-15) is a small protein that was first identified as an abundant phosphoprotein in brain. PEA-15 was characterised so far at the immunochemical level and by a microsequencing attempt. In order to update characterisation of this important structure by advanced methodology unambiguously identifying proteins independent of antibody availability and specificity, we used a proteomic method for this purpose: Performing protein profiling in mouse hippocampi using two dimensional gel electrophoresis with subsequent mass spectrometrical (MS/MS) identification we detected this protein and demonstrate proteomic characterisation of PEA-15 (Q62048). This study enables further specific and unambiguous determination serving as an analytical tool. PMID- 15538644 TI - Effects of ornithine on neutrophil (PMN) free amino acid and alpha-keto acid profiles and immune functions in vitro. AB - The objective of this study was to determine the effects of ornithine on polymorphonuclear leucocyte (PMN) free amino- and alpha-keto acid profiles, superoxide anion (O2-) generation, hydrogen peroxide (H2O2) formation and released myeloperoxidase activity (MPO). Exogenous ornithine significantly increased PMN asparagine, glutamine, aspartate, glutamate, arginine, citrulline, alanine, alpha-ketoglutarate and pyruvate as intracellular ornithine increased. Concerning PMN immune function markers ornithine increased H2O2-generation and MPO activity while O2- -formation was decreased. We believe therefore that ornithine is important for affecting PMN "susceptible free amino- and alpha-keto acid pool" although the mechanisms are not yet clear. This may be one of the determinants in PMN nutrition considerably influencing and modulating PMN host defense capability. PMID- 15538645 TI - Molecular characterization of the group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3. AB - The group II chaperonin from the hyperthermophilic archaeum Pyrococcus horikoshii OT3 (PhCPN) and its functional cooperation with the cognate prefoldin were investigated. PhCPN existed as a homo-oligomer in a double-ring structure, which protected the citrate synthase of a porcine heart from thermal aggregation at 45 degrees C, and did the same on the isopropylmalate dehydrogenase (IPMDH) of a thermophilic bacterium, Thermus thermophilus HB8, at 90 degrees C. PhCPN also enhanced the refolding of green fluorescent protein (GFP), which had been unfolded by low pH, in an ATP-dependent manner. Unexpectedly, functional cooperation between PhCPN and Pyrococcus prefoldin (PhPFD) in the refolding of GFP was not observed. Instead, cooperation between PhCPN and PhPFD was observed in the refolding of IPMDH unfolded with guanidine hydrochloride. Although PhCPN alone was not effective in the refolding of IPMDH, the refolding efficiency was enhanced by the cooperation of PhCPN with PhPFD. PMID- 15538648 TI - The role of HFE mutations on iron metabolism in beta-thalassemia carriers. AB - Hereditary hemochromatosis (HH) is an autosomal recessive disorder of iron metabolism characterized by increased iron absorption and progressive storage resulting in organ damage. HFE gene mutations C282Y and H63D are responsible for the majority of HH cases. A third HFE mutation, S65C, has been associated with the development of a mild form of hemochromatosis. The beta-thalassemia trait is characterized by mild, ineffective erythropoiesis that can induce excess iron absorption and ultimately lead to iron overload. The aim of this study was to evaluate the effect of genetic markers (HFE mutations C282Y, H63D, and S65C) on the iron status of beta-thalassemia carriers. A total of 101 individuals heterozygous for beta-thalassemia and 101 normal control individuals were studied. The allelic frequencies of C282Y (1.5 versus 3.5%), H63D (15.3 versus 18.3%), and S65C (1.0 versus 1.5%) did not differ significantly between beta thalassemia carriers and normal controls. Serum iron (P=0.029) and transferrin saturation (P=0.009) were increased in beta-thalassemia carriers heterozygous for H63D mutation. The number of subjects carrying C282Y or S65C mutations was too low to conclude their effect on the iron status. These results suggest that the beta-thalassemia trait tends to be aggravated with the coinheritance of H63D mutation, even when present in heterozygosity. PMID- 15538649 TI - The combination of optical and electrical energies produces different histological findings from when laser alone is used in leg vein treatment. PMID- 15538650 TI - Nonlinear viscoelastic, thermodynamically consistent, models for biological soft tissue. AB - The mechanical behavior of most biological soft tissue is nonlinear viscoelastic rather than elastic. Many of the models previously proposed for soft tissue involve ad hoc systems of springs and dashpots or require measurement of time dependent constitutive coefficient functions. The model proposed here is a system of evolution differential equations, which are determined by the long-term behavior of the material as represented by an energy function of the type used for elasticity. The necessary empirical data is time independent and therefore easier to obtain. These evolution equations, which represent non-equilibrium, transient responses such as creep, stress relaxation, or variable loading, are derived from a maximum energy dissipation principle, which supplements the second law of thermodynamics. The evolution model can represent both creep and stress relaxation, depending on the choice of control variables, because of the assumption that a unique long-term manifold exists for both processes. It succeeds, with one set of material constants, in reproducing the loading unloading hysteresis for soft tissue. The models are thermodynamically consistent so that, given data, they may be extended to the temperature-dependent behavior of biological tissue, such as the change in temperature during uniaxial loading. The Holzapfel et al. three-dimensional two-layer elastic model for healthy artery tissue is shown to generate evolution equations by this construction for biaxial loading of a flat specimen. A simplified version of the Shah-Humphrey model for the elastodynamical behavior of a saccular aneurysm is extended to viscoelastic behavior. PMID- 15538651 TI - Roles of the acidic lipids sulfoquinovosyl diacylglycerol and phosphatidylglycerol in photosynthesis: their specificity and evolution. AB - Sulfoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG) are lipids with negative charges, distributed among membranes of chloroplasts of plants and their postulated progenitors, cyanobacteria, and also widely among membranes of anoxygenic photosynthetic bacteria. Thus, these acidic lipids are of great interest in terms of their roles in the function and evolution of the photosynthetic membranes. The physiological significance of these lipids in photosynthesis has been examined through characterization of mutants defective in their abilities to synthesize SQDG or PG, and through characterization of isolated thylakoid membranes or photosynthetic particles, the acidic lipid contents of which were manipulated in vitro, for example, on treatment with phospholipase to degrade PG. Responsibility of SQDG or PG has been clarified so far in terms of the structural and/or functional integrity of photosystems I and/or II in cyanobacterial, green algal, and higher plant species. Also implied were distinct levels of the responsibility in the different photosynthetic organisms. Extreme cases involved the indispensability of SQDG for photosynthesis and growth in two prokaryotic, photosynthetic organisms and the contribution of PG to construction of the photosystem-I trimer exclusively in cyanobacteria. Here, roles of these acidic lipids are discussed with a focus on their specificity and the evolution of photosynthetic membranes. PMID- 15538652 TI - Cell wall changes involved in the automorphic curvature of rice coleoptiles under microgravity conditions in space. AB - Seedlings of rice (Oryza sativa L. cv. Koshihikari and cv. Tan-ginbozu) were cultivated on board the Space Shuttle STS-95 mission and changes in the morphology and the cell wall properties of coleoptiles were analyzed. In space, rice coleoptiles showed a spontaneous (automorphic) curvature toward the caryopsis in the elongating region. The angle of automorphic curvature was larger in Koshihikari than in a gibberellin-deficient dwarf cultivar, Tan-ginbozu, and the angle gradually decreased during the growth of coleoptiles in both cultivars. The more quickly expanding convex side of the bending region of the rice coleoptiles showed a greater extensibility of the cell wall than the opposite side. There was a significant correlation between the angle of curvature and the difference in the cell wall extensibility between the convex and the concave sides. Both the levels of the cell wall polysaccharides per unit length of coleoptile and the ratio of high-molecular-mass polysaccharides in the hemicellulose fraction were lower in the convex side than the concave one. Also, the activity of (1-->3),(1-->4)-beta-glucanases in the cell wall was higher in the convex side than the concave one. These results suggest that the uneven modifications of cell wall metabolism bring about the difference in the levels and the molecular size of the cell wall polysaccharides, thereby causing the difference in capacity of the cell wall to expand between the dorsal and the ventral sides, leading to the automorphic curvature of rice coleoptiles in space. The data also suggest the involvement of gibberellins in inducing the automorphic curvature under microgravity conditions. PMID- 15538653 TI - Aerenchyma formation and porosity in root of a mangrove plant, Sonneratia alba (Lythraceae). AB - Aerenchyma gas spaces are important for plants that grow in flooded and anaerobic sites or habitats, because these gas spaces provide an internal pathway for oxygen transport. The objective of this study is to characterize the development of aerenchyma gas spaces and observe the porosity in roots of Sonneratia alba. Tissue at different developmental stages was collected from four root types, i.e. cable root, pneumatophore, feeding root and anchor root, of S. alba. In S. alba, gas space is schizogenously produced in all root types, and increases in volume from the root meristem to mature root tissues. The aerenchyma formation takes place immediately, or 3-5 mm behind the root apex. At first, cortical cells are relatively round in cross sections (near the root apex); they then become two kinds of cells, rounded and armed, which combine together, forming intercellular spaces behind the root apex. The average dimensions of cortical cells increased more than 1.3 times in the vertical direction and over 3.3 times in the horizontal direction. At maturity, aerenchyma gas spaces are long tuberous structures without diaphragms and with numerous small pores on the lateral walls. Within the aerenchyma, many sclereids grow intrusively. Root porosity in all root types ranged from 0-60%. Pneumatophores and cable roots had the highest aerenchyma area (50-60%). PMID- 15538654 TI - A novel feature of structural variegation in leaves of the tropical plant Schismatoglottis calyptrata. AB - We report a novel feature of leaf variegation. As is often the case in tropical forest floor herbs, Schismatoglottis calyptrata leaves feature structural variegation. Examination of leaf anatomy in S. calyptrata revealed a novel feature of structural variegation, which was generated by variation in the spatial arrangement of the adaxial-most tip of the palisade cells. In fully green leaf parts, contact between the adaxial-most tip of the palisade cells and the cone-shaped base of the outer epidermis cells was tight, and palisade cells were arranged radially around each epidermal cell. In dull, grayish-green leaf parts, the contact was loose, and no particular spatial arrangement of palisade cells relative to epidermal cells was observed. This feature of structural variation could be disadvantageous for photosynthesis efficiency in view of the hypothesis that, for rainforest herbs, cone-shaped epidermal cells may function as lenses. However, the high frequency of leaf variegation of S. calyptrata in natural habits suggests that this structural variegation plays an unknown advantageous role. PMID- 15538655 TI - The GAOLAOZHUANGREN2 gene is required for normal glucose response and development of Arabidopsis. AB - Recent studies of glucose (Glc) sensing and signaling have revealed that Glc acts as a critical signaling molecule in higher plants. Several Glc sensing-defective Arabidopsis mutants have been characterized in detail, and the corresponding genes encoding Glc-signaling proteins have been isolated. However, the full complexity of Glc signaling in higher plants is not yet fully understood. Here, we report the identification and characterization of a new Glc-insensitive mutant, gaolaozhuangren2 (glz2), which was isolated from transposon mutagenesis experiments in Arabidopsis. In addition to its insensitivity to Glc, the glz2 plant exhibits several developmental defects such as short stature with reduced apical dominance, short roots, small and dark-green leaves, late flowering and female sterility. Treatment with 4% Glc blocked expression of the OE33 gene in wild-type plants, whereas expression of this gene was unchanged in the glz2 mutant plants. Taken together, our results suggest that the GLZ2 gene is required for normal glucose response and development of Arabidopsis. PMID- 15538656 TI - Kinetic resolution of ketoprofen ester catalyzed by lipase from a mutant of CBS 5791. AB - A biotransformation process was developed for the production of (S)-ketoprofen by enantioselective hydrolysis of racemic ketoprofen ester using the mutant Trichosporon laibacchii strain CBS 5791. A satisfactory result was obtained, in which the E was 82.5, with an ee of 0.94 and a conversion of 0.47 under the optimum hydrolysis conditions [E is enantiomeric ratio, E=ln[1-X(1+ee)]/ln[1-X(1 ee)]; ee is enantiomeric excess, ee=(CS-CR)/(CS+CR): temperature of hydrolysis was 23 degrees C]. The medium used in biotransformation was a mixture of growth broth and biotransformation broth at a ratio of 1:9, the concentration of Tween 80 was 15 g/l, the time of hydrolysis, 72 h. These results are promising for further scale-up. Tween 80 significantly improved lipase enantioselectivity and activity at the optimum concentration. PMID- 15538658 TI - An efficient design for birdcage probes dedicated to small-animal imaging experiments. AB - We are presenting, in this paper, a simple, easily reproducible and easily tuneable design for home-built birdcage probes inducing a linearly polarised B1 field. A simple tuning method is proposed which preserves the induced field homogeneity that characterises in general the birdcage design. This is achieved by adding two extra rings (link rings) that allow one to connect two opposed points on the birdcage end rings via tuning capacitors. All the development stages, from design to practical realisation, are presented, step by step, in order to facilitate experimentalist work. Some images obtained with the birdcage probe are presented, accompanied by the B1 maps in transverse as well as longitudinal planes. Finally, a few suggestions are made for obtaining a circularly polarised magnetic field configuration. PMID- 15538659 TI - Cerebral blood flow response to a hypoxic-ischemic insult differs in neonatal and juvenile rats. AB - To compare the cerebral blood flow (CBF) response to a transient episode of hypoxia-ischemia producing damage in neonatal and juvenile rats. One- and four week-old rats were subjected to unilateral carotid artery occlusion plus hypoxia (8% oxygen). Perfusion MR images were acquired either in sham controls or in hypoxic-ischemic rats before, during, 1 h and 24 h after hypoxia-ischemia. At 24 h post hypoxia-ischemia, T2 maps and histology were used to assess damage. In sham controls, CBF increased twofold between the age of one and four weeks. Reductions in CBF ipsilateral to the occlusion occurred during hypoxia-ischemia followed by a substantial recovery at 1 h post in both age groups. However, contralaterally, hyperemia occurred during hypoxia-ischemia in four-week but not one-week-old rats. Similarly, hyperemia occurred ipsilaterally at 24 h post hypoxia-ischemia in four-week but not one-week-olds, corresponding to the distribution of elevations in T2. Despite CBF differences, extensive cell death occurred ipsilaterally in both age groups. The CBF responses to hypoxia-ischemia and reperfusion differ depending on postnatal age, with hyperemia occurring in juvenile but not neonatal rats. The results suggest a greater CBF responsiveness and differential relationship between post-ischemic vascular perfusion and tissue injury in older compared with immature animals. PMID- 15538660 TI - Long-term stability of cardiac function in normal and chronically failing mouse hearts in a vertical-bore MR system. AB - We previously demonstrated stability of ventricular volumes and cardiac function in normal and in chronically failing mouse hearts in MR systems with a vertical bore magnet for up to 1 h. However, in order to exploit the benefits of an increased magnetic field strength of these MR systems in more time-consuming studies required by, for example MR spectroscopy, we investigated whether cardiac function and ventricular volumes of healthy and infarcted mice would be affected in vertical position over a prolonged period. We applied high-resolution MR cine imaging on an 11.7 T vertical MR system to monitor cardiac functional parameters of normal and chronically failing mouse hearts over a period of 3 h in an upright position, with a temporal resolution of < or =15 min. We monitored left ventricular volumes and cardiac functional parameters in both groups. In normal mice, we detected a decrease of left-ventricular end-systolic volumes by 8 microl and an approximately 23% increase of ejection fraction over time indicating a small but detectable degree of orthostatic dysregulation. Observed changes were more pronounced in mice with heart failure. Despite significant changes in left ventricular volumes and function, absolute values measured for all functional cardiac parameters are consistent with near-physiological conditions. Thus, mice can be studied in high-field MR systems positioned vertically for 3 h. PMID- 15538661 TI - [Health disorders through environmental and professional factors]. PMID- 15538662 TI - [Effects of systemic or topical administration of corticosteroids and vitamin B in patients with olfactory loss]. AB - BACKGROUND: Aim of the present, unblinded, multicentric, open trial was to compare effects of 3 treatments in patients with olfactory dysfunction. METHODS: Treatments included administration of systemic corticosteroids (oral prednisolone), local corticosteroids (mometasone nasal spray), and systemic vitamin B, respectively. A total of 192 patients participated (95 women, 97 men; mean age 56 years). Duration of the smell loss ranged from 1 to 288 months (average 45 months). Olfactory dysfunction was due to infections of the upper respiratory tract (n = 72), sinunasal disease (n = 19), and posttraumatic olfactory loss (n = 10); the largest portion was classified as idiopathic (n = 85); other causes were rare (n = 6). RESULTS: Following systemic administration of corticosteroids improvement of olfactory function was observed (p < 0.001). Similarly, improvement of the sense of smell was found 2 (p = 0.03) and 6 months (p = 0.001) after local administration of mometasone, respectively. In contrast, after administration of vitamin B no significant change of olfactory function was seen after 2 months, while improvement was present after 6 months (p = 0.001). Duration of olfactory dysfunction had no effect on changes of smell function. CONCLUSIONS: These results indicate that improvement of olfactory function is found in olfactory dysfunction of different causes. Adequately controlled, blinded studies are needed to further explore potential effects of the various treatments. PMID- 15538663 TI - [200 years testing hearing disorders with speech, 50 years german speech audiometry -- a review]. AB - FROM THE BEGINNIGS TO CLASSICAL SPEECH TESTS: The need for classification of different degrees of hearing disorders first arose when it seemed possible to treat deafness. Grapengiesser in Berlin 1801 had applied galvanic current to the ears of deaf children and reported some success. Pfingsten in Kiel in 1804 using this method was the first to use speech in diagnosing different degrees of deafness. He divided speech sounds into three classes: vowels, voiced consonants and voiceless consonants. Itard in Paris in 1821 gave a classification of five classes according to which sounds could be perceived, starting from normal speech to thunder and the bang of a gun. Schmalz in Dresden 1846 noted the range within which speech was understood thus introducing the concept of hearing distance. Helmholtz in 1863 had demonstrated that vowels are composed of pure tones. Wolf in Frankfurt 1871 tried to align all speech sounds from the lowest frequency (tongue-R = 16 Hz) to the highest (sh = 4096 Hz) and measured the hearing distance for each sound. Following these suggestions word lists based on the predominant frequencies were compiled in a number of languages including Japanese. SPEECH AUDIOMETRY: This chapter is devoted to Karl Heinz HahIbrock, Freiburg, who was the founder of the German speech audiometry. Hahlbrock followed the American authors of the Psycho-acoustic Laboratory at Harvard, in particular J. P. Egan (1948), using statistical methods for composing lists of words based on the relative frequency of speech sounds and phonetically balanced between the different groups. He finally presented a test comprising groups of two-digit numbers and monosyllabic words. Hahlbrock died in 2003 exactly fifty years after the presentation of his test. A short account of his life is given. THE FOLLOWING DEVELOPMENT: In the following years various other types of speech tests were elaborated using sentences, distorted speech, diotic and dichotic presentation partly aimed at fitting hearing aids, partly with the aim to diagnose central hearing disorders. Hahlbrock's test, however, remained the standard for evaluating speech reception and discrimination. DISCUSSION: One of the fundamental problems in testing speech discrimination is that there is no catalogue of phonemes common to all languages or regional accents. Untrained not native speakers often do no perceive certain sounds having a partial auditory agnosia. They cannot distinguish between e. g. 'hand' and 'and' or 'end' and 'ant', but the examiner must decide if the word presented was repeated correctly or not. PMID- 15538664 TI - [Etiology and therapy of the internal jugular vein thrombosis]. AB - BACKGROUND: Thromboses of the upper limp and neck are rare in comparison with those of the lower extremities. Internal jugular vein thrombosis (IJVT) is a serious event with a potentially fatal outcome. Complications include pulmonary embolism, sepsis with septic emboli to different organs and tissues as well as intracranial propagation of the thrombus with cerebral edema. As any thrombosis, IJVT is precipitated by Virchow's triad: endothelial damage, alteration of blood flow and hypercoagulability. The history and examination in patients with an IJVT may be vague and misleading. Patients may present with a painful swelling of the neck but they may also be absolutely asymptomatic. Imaging procedures frequently used to diagnose an IJVT include sonography with color-coded duplex sonography, computed tomography, magnetic resonance imaging as well as magnetic resonance venography. Up to date, there is no standardized treatment regimen for patients with an IJVT. PATIENTS AND METHODS: This retrospective study includes all ten patients with an IJVT who were seen at our department between January 2000 and January 2004. There were six female and four male patients. The average age was 49.7 years, ranging from 28 to 79 years. RESULTS: In five cases, the thrombosis was associated with a malignant tumor. In four patients, it was caused by a deep neck space infection and in one case the IJVT was due to cervical, intravenous drug abuse. Two patients were found to be pregnant (one tumor patient and one patient with a deep neck space infection). In all cases, a ten day treatment regimen with intravenous antibiotics and anticoagulant therapy was initiated. Oral or subcutaneous anticoagulation was continued for six weeks to six months. No complications were seen in any patient. In three patients a revascularization of the affected vessel could be demonstrated with color-coded duplex sonography six months after the initial presentation. CONCLUSIONS: Thrombosis of the IJV is probably underdiagnosed. Since the clinical presentation may be vague or misleading, a high degree of suspicion is required to make the diagnosis. The potential complications such as pulmonary embolism or intracranial propagation of the thrombus may be fatal. Whenever the thrombosis is not caused by an inflammatory process, a malignant tumor should be excluded. We recommend a therapy with intravenous antibiotics as well as a systemic anticoagulation. Ligation or resection of the internal jugular vein is reserved for patients who develop complications despite adequate medical therapy. PMID- 15538665 TI - [The interesting case -- case no. 65]. AB - Bilateral Osteomyelitis following frontal sinusitis is a rare complication in the antibiotic era. The main risk of a progredient course is mainly the formation of external subperiostal, epidural, subdural abscesses or brain abscesses with potentially life-threatening complications. This is a report on a patient who presented in our department with progredient cephalgia and swelling in the area around the glabella and frontal sinus. Macroscopical examination revealed septum deviation to the right side and nasal polyposis of the right nasal passage. A computed tomography of the paranasal sinuses showed subtotal soft tissue obstruction of all sinuses with the exception of the sphenoidal sinus. Bilateral osteolysis of the ventral osseous borders of the frontal sinus with accompanying osteosclerosis was also observed. The therapy consisted of radical frontal sinus surgery via Unterberger approach. This case demonstrates a rare case of bilateral frontal osteomyelitis together with the necessary diagnostic and therapeutic measures. The management consisted of the removal of all osteomyelitic bone and antibiotic therapy. Differential diagnostic procedures must be carried out in order to exclude orbital and intracranial complications. Regular follow-up examinations and a CT scan of the paranasal sinuses are part of the standard therapy. PMID- 15538666 TI - [Sensible application of modern hearing aids--technic, uses and limitations]. PMID- 15538667 TI - [Conductive hearing and speech-language-delay: correlation, causality and consequences]. PMID- 15538668 TI - [Initial experiences with German DRG case reimbursement system]. PMID- 15538669 TI - [Management of intraoperative complications during middle ear surgery]. PMID- 15538670 TI - [Multiple Chemical Sensitivity (MCS) -- scientific and health policy aspects]. AB - Multiple Chemical Sensitivity (MCS) is characterized by an increased sensitivity towards chemicals. Many patients with severed unspecific symptoms ascribe these to MCS. At present there is neither a generally accepted diagnosis nor a therapeutic concept. The ENT doctor should look at the nasal airflow resistance, nasal mucosa and olfactory system. PMID- 15538671 TI - [Sensitive phase of maturation of the auditory pathways -- parallel to the eye?]. PMID- 15538672 TI - [Early diagnosis of laryngeal cancer]. PMID- 15538673 TI - [Epidemiology of laryngeal cancer]. PMID- 15538674 TI - [The free tissue transplant -- development and present trends]. PMID- 15538675 TI - [The importance of socio-cultural factors for the development of speech and language]. PMID- 15538676 TI - [Management of quality in rehabilitation with hearing aids]. PMID- 15538677 TI - [Rehabilitation concept after laryngectomy]. PMID- 15538678 TI - Pounded in Scotland. PMID- 15538679 TI - The human first carpometacarpal joint: osteoarthritic degeneration and 3 dimensional modeling. AB - The purpose of this study was to gain insight into potential mechanical factors contributing to osteoarthritis of the human first carpometacarpal joint (CMC). This was accomplished by creating three-dimensional (3-D) computer models of the articular surfaces of CMC joints of older humans and by determining their locus of cartilage degeneration. The research questions of this study were: 1) What is the articular wear pattern of cartilage degeneration in CMC osteoarthritis?, (2) Are there significant topographic differences in joint area and contour between the joints of males and females?, and 3) Are there measurable bony joint recesses consistently found within the joint? The articular surfaces of 25 embalmed cadaveric joints (from 13 cadavers) were graded for degree of osteoarthritis, and the location of degeneration was mapped using a dissection microscope. The surfaces of 14 mildly degenerated joints were digitized and reconstructed as 3-D computer models using the Microscribe 3D-X Digitizer and the Rhinoceros 2.0 NURBS Modeling Software. This technology provided accurate and reproducible information on joint area and topography. The dorsoradial trapezial region was found to be significantly more degenerated than other quadrants in both males and females. Mean trapezial articular surface area was 197 mm 2 in males and 160 mm(2) in females; the respective mean areas for the metacarpal were 239 mm(2) in males and 184 mm(2) in females. Joints of females were found to be significantly more concave in radioulnar profile than those of males. Three bony joint recesses were consistently found, two in the radial and ulnar aspects of the trapezium and the third in the palmar surface of the metacarpal. PMID- 15538680 TI - Splinting the degenerative basal joint: custom-made or prefabricated neoprene? AB - The purpose of this study was to compare the objective, subjective, and radiographic responses of patients with carpometacarpal joint osteoarthritis (CMCJ-OA) wearing a prefabricated neoprene splint (PFN), which crosses the CMCJ and metacarpophalangeal joint, with those of patients wearing a custom-made thermoplastic short opponens splint (CMT), which crosses only the CMCJ. Patients ( N = 25) with first CMCJ stage I and II osteoarthritis were assigned randomly to wear either the PFN splint or the CMT splint for one week. After one week, the subjects rated their function in the splint and their satisfaction and pain levels on visual analogue scales. Pinch measurements were performed and x-rays were taken to assess carpometacarpal subluxation. The second splint was then applied for one week and all measures were repeated. The subjects rated the PFN splint significantly higher, and most reported that they would choose the PFN splint over the CMT splint for daily and long-term use. Both pain and function were improved with splinting, but the effect was amplified with the PFN splint compared with the CMT splint. Both splints reduced subluxation at the first carpometacarpal joint, but the CMT effect was greater. This study further supports current evidence that subjects with stage I and II first CMCJ-OA will have pain relief with thumb splinting. In addition, the PFN splint will provide greater relief when compared with the CMT splint. Furthermore, this study reveals that patients prefer the PFN splint to the CMT splint. PMID- 15538681 TI - A five-year followup of hand function and activities of daily living in systemic sclerosis (scleroderma). AB - The purpose of this study was to identify hand factors that change over a five year period that may be risk factors for the development of functional disability in persons with systemic sclerosis (scleroderma). Sixty individuals with scleroderma were administered assessments of grip and pinch strength, joint range of motion, and pain, and were observed for the presence of digital ulcers, digital scars, calcium deposits, puffy fingers, and tendon friction rubs. Matched pairs chi square analyses and Fisher's exact tests were performed to compare variables at year 1 and five years later. Grip and pinch strength increased as did joint motion except for the wrist and thumb carpometacarpal joint. There were also significant increases in the presence of scars, friction rubs, calcium deposits, and puffy fingers. Regression analysis was done to determine which variables predicted functional ability. Only puffy fingers predicted functional disability. In conclusion, hand impairment persisted over time while functional ability decreased. PMID- 15538682 TI - Determining the accuracy of the psychophysical approach to grip force measurement. AB - This study examined the accuracy of estimating actual grip force by measuring perceived force simulation using a hand dynamometer. A multifactorial design was used to ascertain whether there was a difference between actual and perceived grip force performance using three levels of force, three different age groups, males and females, and both dominant and nondominant hands. One hundred ninety eight participants were asked to exert an exact amount of grip force marked on a dynamometer, and then asked to replicate the same amount without the visual feedback of the dial. This was repeated for both hands for all three force levels. Significant differences were found between the actual and perceived grip force performance using three different levels of force, among the age categories, and between males and females. This study provides evidence that use of the psychophysical method of grip force measurement may not be an accurate methodology for determining task demands involving grip force. PMID- 15538683 TI - Evaluation of the Chinese version of the Disability of the Arm, Shoulder and Hand (DASH-HKPWH): cross-cultural adaptation process, internal consistency and reliability study. AB - The Disability of the Arm, Shoulder and Hand (DASH) was translated into Chinese by a physiotherapy team of the Prince of Wales Hospital, Hong Kong (DASH-HKPWH). OBJECTIVES: This study evaluated the cross-cultural adaptation process, face validity, internal consistency and reliability of the DASH-HKPWH. METHOD: Language officers and medical professionals from different fields were invited to translate and evaluate the face validity of the DASH-HKPWH. 88 patients were recruited to complete two DASH questionnaires on two occasions 1-2 weeks apart. RESULTS: Some adjustments were made to the translations based on the cultural and linguistic practice in Hong Kong. The face validity was satisfactory with a mean endorsement score of 3.2. The difference between the mean of DASH scores was not significant (t = -0.35, p = 0.73). The ICC (1,1) and Cronbach's alpha for the 30 item Disability/Symptom of the DASH-HKPWH was 0.77 and 0.94, respectively. CONCLUSION: The translation was valid and reliable and acceptably equivalent to the original version. The questionnaire is suitable for measuring changes experienced by patients with any upper extremity disorders. PMID- 15538684 TI - A double reverse Kleinert extension splint for extensor tendon repairs in zones VI to VIII. PMID- 15538706 TI - Evidence for the structure of the enantioactive ligand in the phosphine-copper catalyzed addition of diorganozinc reagents to imines. PMID- 15538707 TI - Molecular characteristics of Escherichia coli serogroup O78 strains isolated from diarrheal cases in bovines urge further investigations on their zoonotic potential. AB - We investigated the virulence properties and clonal relationship of 21 Escherichia coli strains of serogroup O78 isolated from diarrhoeic cattle and calves. Isolates were screened for 18 genes representing virulence features of different Escherichia coli pathotypes. None of the strains harboured enterotoxin genes estIa/Ib, eltIa/Ib, or Shiga toxin (stx) genes, genes involved in adhesion (eae, f5, f41) hemolysin gene hlyA or invasion gene ipaC. With a high prevalence we detected enterotoxin astA (61.9%), genes involved in iron acquisition, like fyuA, irp (each 57.1%) and iucD (81.0%), and the operon sequence of Colicin V plasmids (38.1%). Some strains possessed toxin genes cdt-IIIB and cnf1/2 (both 14.3%), the invasion gene tia (23.8%), and the serine protease encoding gene espP (23.8%). Moreover, we could show that E. coli O78 strains under investigation were able to adhere to and invade MDBK-cells with varying efficiencies. The results indicate that the closely related O78 strains, constituting two major PFGE-clusters, harbor various virulence features for bovine intestinal disease but cannot be grouped into one of the common E. coli intestinal pathogenic or other pathotypes according to their virulence gene pattern. Nevertheless, the ability to adhere, invade or harbor toxin genes lets us suggest that O78 strains isolated from diarrheal cases in bovines urges further investigations on the zoonotic potential of these strains. PMID- 15538708 TI - First observation of two consecutive gamma turns in a crystalline linear dipeptide. PMID- 15538709 TI - Bacillus cereus, the causative agent of an emetic type of food-borne illness. AB - Bacillus cereus is the causative agent of two distinct forms of gastroenteritic disease connected to food-poisoning. It produces one emesis-causing toxin and three enterotoxins that elicit diarrhea. Due to changing lifestyles and eating habits, B. cereus is responsible for an increasing number of food-borne diseases in the industrial world. In the past, most studies concentrated on the diarrhoeal type of food-borne disease, while less attention has been given to the emetic type of the disease. The toxins involved in the diarrhoeal syndrome are well known and detection methods are commercially available, whereas diagnostic methods for the emetic type of disease have been limited. Only recently, progress has been made in developing identification methods for emetic B. cereus and its corresponding toxin. We will summarize the data available for the emetic type of the disease and discuss some new insights in emetic strain characteristics, diagnosis, and toxin synthesis. PMID- 15538710 TI - A rapid method for the discrimination of genes encoding classical Shiga toxin (Stx) 1 and its variants, Stx1c and Stx1d, in Escherichia coli. AB - Subtyping of Shiga toxin (Stx)-encoding genes by conventional polymerase chain reaction (PCR) is time-consuming. We developed a single step real-time fluorescence PCR with melting curve analysis to distinguish rapidly stx1 from its variants, stx1c and stx1d. Melting temperatures (Tm) of 206 Stx-producing Escherichia coli (STEC) identified to harbor stx1 or stx1c were analyzed using a specific hybridization probe over the variable region. 170 of 171 stx1-harboring STEC displayed Tm of 69 degrees C to 70 degrees C, whereas 34 of 35 strains containing stx1c had Tm of 65 degrees C-66 degrees C. This constant and reproducible difference of 4 degrees C demonstrated that melting curve analysis is a reliable technique to differentiate stx1 from stx1c. Two isolates displayed atypical Tm. Sequence analysis showed that one of them was 100% identical to stx1d within a 511 bp DNA stretch. Our data demonstrate that real-time PCR is a rapid and reliable tool to differentiate stx1 from stx1c and stx1d and to detect new stx1 variants. Because stx1-harboring STEC cause diarrhoea and hemolytic uremic syndrome, whereas those containing stx1c are often shed asymptomatically, a rapid differentiation between stx1 and its variants using the procedure developed here has both clinical implications and a direct significance for the risk assessment analysis of STEC isolated from foods. PMID- 15538711 TI - The residue levels of narasin in eggs of laying hens fed with unmedicated and medicated feed. AB - Laying hens were fed contaminated feed containing narasin 2.5 mg/kg for 21 days followed by a 7 day withdrawal period, hens in the control group were fed unmedicated feed. Eggs were collected during trial days 0, 3, 7, 14, 21 and after the withdrawal period of 7 days. The concentration of narasin in yolks and egg whites was analyzed by a liquid chromatography-mass spectrometry method. Narasin was found to accumulate in yolks, where the narasin concentration increased during the treatment. The concentration of narasin varied from 5.9 to 13.8 microg/kg (mean 10.6 microg/kg) in yolks after 21 day feeding periods. The concentrations of narasin ranged from < 0.9 to 1.4 microg/kg after the withdrawal period. Narasin residues were not found in egg whites of the laying hens fed contaminated feed nor in either yolks or egg whites of the laying hens fed unmedicated feed. The effect of cooking was also tested on the amount of narasin residues in eggs. Cooking for 10 min did not significantly influence the narasin residues in eggs. Traces of lasalocid were also found in the yolks. The traces of lasalocid are attributable to an accidental contamination of the feed during its manufacture. PMID- 15538712 TI - Expression profiles of effector proteins SopB, SopD1, SopE1, and AvrA differ with systemic, enteric, and epidemic strains of Salmonella enterica. AB - The presence and expression of sopB, sopD1, sopE1, and avrA genes encoding virulence associated effector proteins were studied comparatively in 405 Salmonella enterica strains. They belong to different serovars and clonal types (genotypes, phage types) and originated from different clinical (systemic infection, focal enteritis, enterocolitis) and epidemic sources (epidemics, sporadic cases). The sopB and sopD1 determinants were commonly prevalent, but sopE1 and avrA genes only in 55% and 80%, respectively. A correlation of this pattern of absence and presence of the respective genes to the epidemic and clinical origin could not be detected. In contrast, the expression of the respective genes appeared differently: SopB and SopE1 proteins are well produced, but SopD1 and AvrA proteins only rarely under the applied standard culture conditions. However, using a range of different environmental signals (temperature, pH, cations, etc.) some of the S. enterica nonproducer strains (e. g., S. Agona, S. Bovismorbificans, S. Virchow, etc.) begin to produce AvrA and SopD1. They turned now into an expression profile which was found typically for the epidemic strains of S. Typhimurium and S. Enteritidis. Also S. enterica strains from systemic infections could be characterized by their strong SopB and SopE1 expression while SopD1 and AvrA proteins were missing. Although it is premature to outline generally a correlation of these expression profiles and the clinical and epidemiological potency of Salmonellae, the reported results allow a first understanding how a fine tuning of their virulence will take place. PMID- 15538713 TI - Molecular monitoring of environmental conditions influencing the induction of ochratoxin A biosynthesis genes in Penicillium nordicum. AB - A real-time polymerase chain reaction (PCR) system specific for the ochratoxin A polyketide synthase gene (otapksPN) of Penicillium nordicum has been used to analyze environmental conditions, influencing the induction of that key gene of the ochratoxin A biosynthetic pathway. Generally, the induction of that gene coincides very well with the biosynthesis of ochratoxin A, demonstrating that its induction can be used as a molecular signal to monitor ochratoxin A production. It could be shown, that the expression of the otapksPN gene is greatly dependent on the media used. In YES medium expression is highest, followed by minimal medium which support ochratoxin A production and minimal medium which suppresses ochratoxin A production. The amount of ochratoxin A produced shows the same tendency. The amount produced is highest on YES medium and decreases successively to the two minimal media. The system was also used to determine the influence of environmental parameters like temperature, pH and NaCl concentration on the expression of the otapksPN gene and on ochratoxin A production in parallel. It could be shown that under acidic conditions, below pH 5.0, the expression of the otapksPN gene as well as the ochratoxin A concentration were reduced. In case of salt concentration again both measures coincide, having both highest values at increasing NaCl concentrations. In case of the temperature, however, expression of the otapksPN gene was uncoupled to ochratoxin A production. The expression was high at all temperatures tested, however, clear differences in the biosynthesis of ochratoxin A by P. nordicum could be observed at the different temperatures, showing highest production at 25 degrees C. The importance of these data are discussed with reference to the natural habitat of P. nordicum. PMID- 15538714 TI - Occurrence of antibiotic-resistant enterobacteria in agricultural foodstuffs. AB - Antibiotic-resistant bacteria or their corresponding resistance determinants are known to spread from animals to humans via the food chain. We screened 20 vegetable foods for antibiotic-resistant coliform bacteria and enterococci. Isolates were directly selected on antibiotic-containing selective agar (color detection). Thirteen "common vegetables" (tomato, mushrooms, salad) possessed 10(4)-10(7) cfu/g vegetable of coliform bacteria including only few antibiotic resistant variants (0-10(5) cfu/g). All seven sprout samples showed a some orders of magnitude higher contamination with coliform bacteria (10(7)-10(9) cfu/g) including a remarkable amount of resistant isolates (up to 10(7) cfu/g). Multiple resistances (up to 9) in single isolates were more common in sprout isolates. Resistant bacteria did not originate from sprout seeds. The most common genera among 92 isolates were: 25 Enterobacter spp. (19 E. cloacae), 22 Citrobacter spp. (8 C. freundii), and 21 Klebsiella spp. (9 K. pneumoniae). Most common resistance phenotypes were: tetracycline (43%), streptomycin (37%), kanamycin (26%), chloramphenicol (29%), co-trimoxazol (9%), and gentamicin (4%). The four gentamicin-resistant isolates were investigated in molecular details. Only three (chloramphenicol) resistant, typical plant-associated enterococci were isolated from overnight enrichment cultures. In conclusion, a contribution of sprouts contaminated with multiresistant, Gram-negative enterobacteria to a common gene pool among human commensal and pathogenic bacteria cannot be excluded. PMID- 15538715 TI - Molecular and cellular effects of green tea on oral cells of smokers: a pilot study. AB - Studies in cell culture and laboratory animals have shown that green tea and its major component, epigallocatechin-3-gallate, inhibit cell growth and reduce tumor incidence. However, results of epidemiological studies have generated inconsistent, sometimes conflicting data regarding protection by green tea against human cancers. To clarify the findings of these laboratory studies in application to humans, we conducted a pilot intervention study with three heavy smokers (> 10 cigarettes/day) and three nonsmokers (never smokers) in order to evaluate the molecular and cellular effects of drinking green tea using human oral cells as an investigative tool. Green tea total extract (400-500 mg/cup, 5 cups/day) was administered in drinking water to the subjects for four weeks. Two oral cytology samples were taken weekly for measurements of tobacco carcinogen induced DNA damage, including bulky adducts and oxidized bases, cell growth, DNA content, and apoptosis. The study showed that during the course of green tea administration smoking-induced DNA damage was decreased, cell growth was inhibited, and the percentage of cells in S phase was reduced, cells accumulated in G1 phase (cyclin D1 positive), DNA content became more diploid and less aneuploid, and p53, Caspase-3, and TUNEL, markers of apoptosis, were increased. The study, although preliminary, indicates that drinking green tea reduced the number of damaged cells in smokers by inducing cell growth arrest and apoptosis, a mechanism similar to that observed in cultured cells and animals. These results warrant a large-scale intervention trial to further verify the role of green tea in the prevention of oral cancer in smokers. PMID- 15538716 TI - Micro/nanoscale mechanical and tribological characterization of SiC for orthopedic applications. AB - Micro/nanomechanical and tribological characterization of SiC has been carried out. For comparison, measurements on SiC, CoCrMo, Ti-6Al-4V, and stainless steel have also been made. Hardness and elastic modulus of these materials were measured by nanoindentation using a nanoindenter. The nanoindentation impressions were imaged using an atomic force microscope (AFM). Scratch, friction, and wear properties were measured using an accelerated microtribometer. Scratch and wear damages were studied using a scanning electron microscope (SEM). It is found that SiC exhibits higher hardness, elastic modulus, scratch resistance as well as lower friction with fewer and smaller debris particles compared to other materials. These results show that SiC possesses superior mechanical and tribological properties that make it an ideal material for use in orthopedic and other biomedical applications. PMID- 15538717 TI - Towards gelsolin amyloid formation. AB - Amyloid diseases result from protein misfolding and aggregation into fibrils. Some features of gelsolin amyloidogenic fragments comprised of residues 173-243 (G173-243) and residues 173-202 (G173-202) were investigated by the method of molecular dynamics (MD). The alpha-helical structure of G173-243 present in the whole protein unwinds during the course of MD simulation of the fragment G173 243, suggesting that the G173-243 structure is not stable and could unfold before becoming involved in gelsolin amyloid fibril formation. Twelve fragments of G173 202 were used to build a possible beta-fibril. During the course of the simulation, G173-202 fragments formed hydrogen bonds and tended to turn by an angle of 10 degrees -20 degrees towards each other. PMID- 15538718 TI - Preferential humoral immune response in prostate cancer to cellular proteins p90 and p62 in a panel of tumor-associated antigens. AB - BACKGROUND: Cytoplasmic p90 autoantigen was recently cloned from a cDNA expression library using serum antibody from a cancer patient. The humoral immune response to p90 in prostate cancer and benign prostatic hyperplasia (BPH) was examined. METHODS: An antigenic fragment of recombinant p90 protein and several other tumor-associated antigens (TAAs) were used in ELISA and Western blotting to detect antibodies in sera from patients with prostate cancer, BPH, and other controls. RESULTS: Autoantibodies to p90 were detected in 30.8% of 133 prostate cancer patients versus 1.5% in 68 BPH patients. When a selected panel of six TAAs including p90 were used for immunoscreening, the cumulative positive reactions in prostate cancer sera reached 92.5%, significantly higher than in BPH and other control sera. Antibodies to p90 showed the highest frequency in prostate cancer (30.8%), followed by antibodies to p62 (22.6%). CONCLUSIONS: A panel of six selected TAAs was shown to have high sensitivity and specificity as immunodiagnostic markers in prostate cancer. PMID- 15538719 TI - Truncated E-cadherin potentiates cell death in prostate epithelial cells. AB - BACKGROUND: E-cadherin, a fundamental component of the adherens junction, is known to mediate aggregation-dependent cell survival. We have previously identified a novel, calpain-dependent proteolytic cleavage of E-cadherin that resulted in the generation of a stable 100-kDa E-cadherin fragment (E-cad(100)) in prostate epithelial cells in response to cell death stimuli. We postulated that the E-cad(100) fragment may play a role in abrogating survival of LNCaP cells following induction of apoptosis. METHODS: Wild-type E-cadherin and E cad(100) were engineered, tagged with GFP, and stably expressed in LNCaP cells. These cell lines were characterized for E-cadherin-GFP/beta-catenin interactions, endogenous E-cadherin and beta-catenin expression, and sensitivity to apoptosis induced by PKC activation. RESULTS: E-cad(100)-GFP demonstrated a punctuate expression pattern, in contrast to E-cad(120)-GFP, which was membrane-localized. E-cad(100)-GFP, unlike E-cad(120)-GFP, failed to bind to and co-localize with beta-catenin. Transient or stable overexpression of E-cad(100) resulted in the downregulation of endogenous E-cadherin expression at the cell membrane. Activation of PKC in LNCaP cells which overexpressed E-cad(100) potentiated cell death. CONCLUSIONS: Truncated E-cadherin may play a role in the regulation of endogenous E-cadherin expression and epithelial cell survival. PMID- 15538720 TI - Stimulating in-soil rhamnolipid production in a bioslurry reactor by limiting nitrogen. AB - A soil with aged contamination from lubricating oil (LO) and polychlorinated biphenyls (PCBs) was treated in a bioslurry reactor to investigate in-soil biosurfactant production by Pseudomonas aeruginosa, the most abundant indigenous, culturable, hydrocarbon-degrading microorganism. After 2 days of growth on LO, a depletion of nitrogen stimulated the production and accumulation of rhamnolipids to levels roughly 20 times the critical micelle concentration. Surface tensions and concentrations of monorhamnolipid and dirhamnolipid, PCBs, and total petroleum hydrocarbons (TPH) were measured in a slurry filtrate. Soil-bound PCBs and TPH were also quantified. Rhamnolipid production was observed within 1 to 2 days after nitrogen depletion in each of the 10 batches tested. By day 6, total rhamnolipid concentrations increased from below detection to average values over 1,000 mg/L, which caused over 98% of soil-bound PCBs and over 99% of TPH to be emulsified and recovered in the filtrate. After 70 days, rhamnolipid concentrations were only reduced by 15%, because of nitrogen-limited rates of rhamnolipid biodegradation. The results show that in-soil biosurfactant production can be stimulated in a controlled way with nutrient limitation and can be used to achieve soil washing. PMID- 15538721 TI - Toward an aggregated understanding of enzymatic hydrolysis of cellulose: noncomplexed cellulase systems. AB - Information pertaining to enzymatic hydrolysis of cellulose by noncomplexed cellulase enzyme systems is reviewed with a particular emphasis on development of aggregated understanding incorporating substrate features in addition to concentration and multiple cellulase components. Topics considered include properties of cellulose, adsorption, cellulose hydrolysis, and quantitative models. A classification scheme is proposed for quantitative models for enzymatic hydrolysis of cellulose based on the number of solubilizing activities and substrate state variables included. We suggest that it is timely to revisit and reinvigorate functional modeling of cellulose hydrolysis, and that this would be highly beneficial if not necessary in order to bring to bear the large volume of information available on cellulase components on the primary applications that motivate interest in the subject. PMID- 15538722 TI - Dynamic effects related to steady-state multiplicity in continuous Saccharomyces cerevisiae cultivations. AB - The behavioral differences between chemostat and productostat cultivation of aerobic glucose-limited Saccharomyces cerevisiae were investigated. Three types of experiments were conducted: a chemostat, where the dilution rate was shifted up or down in stepwise manner; and a productostat, with either stepwise changed or a rampwise increased ethanol setpoint, i.e., an accelero-productostat. The transient responses from chemostat and productostat experiments were interpreted using a simple metabolic flux model. In a productostat it was possible to obtain oxido-reductive steady states at dilution rates far below Dcrit due to a strong repression of the respiratory system. However, these steady states could not be obtained in a chemostat, since a dilution rate shift-down from an oxido-reductive steady state led to a derepression of the respiratory system. It can therefore be concluded that the range of dilution rates where steady-state multiplicity can be obtained differs depending on the operation mode and that this dilution rate multiplicity range may appear larger in a productostat than in a chemostat. A more narrow multiplicity range, however, was obtained when the productostat was operated as an accelero-productostat. PMID- 15538723 TI - Osteosarcoma cell lines display variable individual reactions on wildtype p53 and Rb tumour-suppressor transgenes. AB - BACKGROUND: One of the most widely studied gene therapeutic strategies for cancer is the introduction of tumour-suppressor genes-generally p53-into the target cells. As the genes of p53 and/or retinoblastoma (Rb) are mutated in the major part of osteosarcomas (OS), we aimed to study the effect of p53 and Rb transgenes on a panel of five different osteosarcoma cell lines. METHODS: OS cell lines were transduced by adenoviral vectors delivering the transcription units of the wildtype p53 and the Rb gene. Effects of the transgenes alone and at additional cytostatic stress were studied by proliferation, alive/dead and cell cycle assays. RESULTS: The individual cells lines displayed divergent reactions to p53- or Rb-transgene delivery reaching from cell death (SaOs-2, U2OS at p53 transduction) over stopped or lowered cell division (MG-63, K-HOS, SJSA-1 at p53 and Rb transduction) to nearly unhindered cell growth (U2OS at Rb transduction). In those OS cell lines reacting with lowered cell division to p53 or Rb delivery, cytostatics only moderately intensified the transgene effects. Surprisingly, these reactions were apparently not dependent on the functional status of the cellular p53 and/or Rb genes or on differences in the infectability of the cell lines by the adenoviral vectors. Most interestingly, the respective effects of the p53 or Rb transgenes were not multiplied by simultaneous transduction of both tumour-suppressor genes. CONCLUSIONS: The application of wildtype tumour suppressor gene therapy on genetically variable osteosarcomas may be efficient only in yet not identified genetic subgroups of this tumour entity. Hyperactive tumour-suppressor transgenes could be an alternative. PMID- 15538724 TI - A novel chimeric ribozyme vector produces potent inhibition of ICAM-1 expression on ischemic vascular endothelium. AB - BACKGROUND: Inhibition of intercellular adhesion molecule-1 (ICAM-1) expression can ameliorate the inflammation induced by ischemia-reperfusion injury (IRI) in animal models. However, current strategies to reduce ICAM-1 expression have been limited by the lack of stability, poor specificity, and the transient nature of synthesized regulatory molecules (antisense/ribozyme). METHODS: A chimeric expression vector was generated by fusing a ribozyme targeting sequence against ICAM-1 to stabilizing stem-loop structures and nuclear localization signals that are components of endogenous U1 small nuclear RNA. Oligonucleotide scanning was used to predict accessible sites for targeting within the rat ICAM-1 transcript. Efficacy of the chimeric ribozyme vector was tested by transfection of rat aortic endothelial (RAE) cells (in vitro) and intraportal delivery in a rat hepatic IRI model (in vivo). RESULTS: Transfection of RAE cells with the chimeric ribozyme vector produced potent and specific inhibition of ICAM-1 mRNA and protein levels by >65%. This reduction in ICAM-1 expression was accompanied by a proportional decrease in neutrophil adhesion to RAE cells. In vivo intraportal delivery of the chimeric targeting vector to rats sustaining hepatic IRI produced a marked reduction in ICAM-1 expression on liver endothelium after reperfusion. CONCLUSIONS: A chimeric ribozyme vector effectively inhibited ICAM-1 expression in vascular endothelial cells and in rat liver following IRI, demonstrating a novel gene targeting technique that may be ideally suited to clinical applications aimed at ameliorating IRI. PMID- 15538725 TI - The utrophin promoter A drives high expression of the transgenic LacZ gene in liver, testis, colon, submandibular gland, and small intestine. AB - BACKGROUND: Duchenne muscular dystrophy (DMD) is caused by the absence of the muscle cytoskeletal protein dystrophin. Utrophin is an autosomal homologue of dystrophin, and overexpression of the protein is expected to compensate for the defect of dystrophin. The utrophin gene has two promoters, A and B, and promoter A of the utrophin gene is a possible target of pharmacological interventions for DMD because A-utrophin is up-regulated in dystrophin-deficient mdx skeletal and cardiac muscles. To investigate the utrophin promoter A activity in vivo, we generated nuclear localization signal-tagged LacZ transgenic mice, where the LacZ gene was driven by the 5-kb flanking region of the A-utrophin gene. METHODS: Four transgenic lines were established by mating four independent founders with C57BL/6J mice. The levels of mRNA for beta-galactosidase in several tissues were examined by RT-PCR. Cryosections from several tissues were stained with hematoxylin and eosin (H&E) and with 5-bromo-4-chloro-3-indolyl-beta-D galactopyranoside (X-Gal). RESULTS: The 5-kb upstream region of the A-utrophin gene showed high transcriptional activity in liver, testis, colon, submandibular gland, and small intestine, consistent with the endogenous expression of utrophin protein. Surprisingly, the levels of both beta-gal protein and mRNA for the transgene in cardiac and skeletal muscles were extremely low, even in nuclei near the neuromuscular junctions. These results indicate that the regulation of the utrophin gene in striated muscle is different from that in non-muscle tissues. CONCLUSIONS: Our results clearly showed that the utrophin A promoter is not sufficient to drive expression in muscle, but other regulatory elements are required. PMID- 15538726 TI - Deficiency of oncoretrovirally transduced hematopoietic stem cells and correction through ex vivo expansion. AB - BACKGROUND: Extensive efforts to develop hematopoietic stem cell (HSC) based gene therapy have been hampered by low gene marking. Major emphasis has so far been directed at improving gene transfer efficiency, but low gene marking in transplanted recipients might equally well reflect compromised repopulating activity of transduced cells, competing for reconstitution with endogenous and unmanipulated stem cells. METHODS: The autologous settings of clinical gene therapy protocols preclude evaluation of changes in repopulating ability following transduction; however, using a congenic mouse model, allowing for direct evaluation of gene marking of lympho-myeloid progeny, we show here that these issues can be accurately addressed. RESULTS: We demonstrate that conditions supporting in vitro stem cell self-renewal efficiently promote oncoretroviral mediated gene transfer to multipotent adult bone marrow stem cells, without prior in vivo conditioning. Despite using optimized culture conditions, transduction resulted in striking losses of repopulating activity, translating into low numbers of gene marked cells in competitively repopulated mice. Subjecting transduced HSCs to an ex vivo expansion protocol following the transduction procedure could partially reverse this loss. CONCLUSIONS: These studies suggest that loss of repopulating ability of transduced HSCs rather than low gene transfer efficiency might be the main problem in clinical gene therapy protocols, and that a clinically feasible ex vivo expansion approach post-transduction can markedly improve reconstitution with gene marked stem cells. PMID- 15538727 TI - Methodological optimization of polyethylenimine (PEI)-based gene delivery to the lungs of mice via aerosol application. AB - BACKGROUND: Polyethylenimine (PEI) has been successfully used for gene delivery to the lungs of mice via aerosol application using a whole body nebulization device. In this report we optimized the design of such an aerosol device. METHODS: Aerosol devices were constructed as either serial inhalation apparatus or as a whole body nebulization chamber connected to an aerosol spacer placed in a horizontal or vertical position. PEI-based gene vectors were nebulized using a standard jet nebulizer and luciferase gene expression of various tissues was examined. RESULTS: Using a whole body aerosol device resulted in luciferase gene expression in the lungs of mice at the same level as compared with a serial inhalation apparatus. Whereas gene expression was enhanced in the presence of 5% CO(2)-in-air, anesthesia of mice strongly decreased gene expression even when mice were intubated with an intravascular cannula. Reduction of the median mass aerodynamic diameter (MMAD) of the aerosol from 3.4 to 0.27 microm by interposition of an aerosol spacer increased gene expression significantly 3 fold. Drying of the aerosol by silica gel additionally increased gene delivery significantly 3-fold. Reporter gene expression mediated by branched PEI 25 kDa was 9- and 15-fold higher as compared with linear PEIs of 22 and 25 kDa, respectively, and was dependent on the DNA concentration. Gene expression was detectable as soon as 6 h after gene vector application and reached a maximum after 72 h but was still detectable after 14 days. The presence of Zn(2+) did not increase gene expression. CONCLUSION: We propose aerosol drying as a novel and simple method of optimizing PEI-based gene delivery to the lungs. PMID- 15538728 TI - The DNA minor groove binding agents Hoechst 33258 and 33342 enhance recombinant adeno-associated virus (rAAV) transgene expression. AB - BACKGROUND: Recombinant adeno-associated viruses (rAAV) are commonly used in pre clinical and clinical gene transfer studies. However, the relatively slow kinetics of rAAV transgene expression complicates in vitro and in vivo experiments. METHODS: 293 and COS-1 cells were transduced with rAAV2-EGFP, rAAV1 EGFP, or rAAV5-EGFP. The rAAV-EGFP expression was analyzed in the presence of Hoechst 33 258 or 33 342 as a function of time and concentration by flow cytometry and fluorescent microscope. Effects of Hoechst on cell cycle populations were determined by flow cytometry. Enhanced green fluorescent protein (EGFP) expression plasmids with or without AAV inverted terminal repeats (ITR) were constructed and gene expression by transient transfection was compared in the presence of Hoechst. RESULTS: We found that Hoechst 33 258 and 33 342 increase both the level and the population of EGFP gene expressing cells, transduced by several different serotypes of rAAV-EGFP. The augmentation of rAAV EGFP expression occurs in different cell types in a concentration-dependent manner. In addition, the Hoechst 33 258 or 33 342 mediated enhancement of rAAV gene expression correlated with an increase of cells in S phase and G2/M phases of the cell cycle. Finally, gene expression from transfected ITR-containing plasmid DNA was also enhanced by Hoechst dyes. CONCLUSIONS: Our results revealed that two different, although related, DNA-binding drugs, Hoechst 33 258 and 33 342, accelerate the kinetics of rAAV transgene expression. These findings may provide the basis for more sensitive assessment of rAAV biological activity and also extend the applications of rAAV for in vivo gene transfer. PMID- 15538729 TI - Development and characterization of a cell line for large-scale, serum-free production of recombinant adeno-associated viral vectors. AB - BACKGROUND: One of the major limitations to the use of adeno-associated virus (AAV) vectors for gene therapy has been the difficulty in producing enough vector to supply a clinical trial. More than 20 000 roller bottles may be required to generate AAV by the traditional transient transfection process to treat 50 patients. A scalable AAV producer cell line grown in serum-free media will meet the needs for the manufacture of AAV gene therapeutics. METHODS: A packaging cell line was generated by introducing the AAV rep and cap genes into A549 cells. From this packaging cell line, a number of producer cell lines were generated by infecting the packaging cell with the appropriate AAV vector. Producer cell lines were then adapted to serum-free suspension conditions for growth in bioreactors. RESULTS: We report here the development of six AAV producer cell lines that generate > 10(4) particles/cell. The rAAV vector preparations from these cell lines have physical and functional characteristics similar to rAAV vectors prepared by transient transfection. To enable large-scale production, producer cell lines were adapted to serum-free suspension and we demonstrate production of AAV at the 15 L scale. In addition, vector preparations from these cell lines were shown to be free of wild-type AAV. CONCLUSIONS: AAV producer cell lines can be readily scaled to meet the needs of clinical trials. One 500 L bioreactor of these producer cells can produce the equivalent of 2500 high capacity roller bottles or 25 000 T-175 tissue culture flasks. PMID- 15538731 TI - DNA vaccination against tumors. AB - DNA vaccines have been used to generate protective immunity against tumors in a variety of experimental models. The favorite target antigens have been those that are frequently expressed by human tumors, such as carcinoembryonic antigen (CEA), ErbB2/neu, and melanoma-associated antigens. DNA vaccines have the advantage of being simple to construct, produce and deliver. They can activate all arms of the immune system, and allow substantial flexibility in modifying the type of immune response generated through codelivery of cytokine genes. DNA vaccines can be applied by intramuscular, dermal/epidermal, oral, respiratory and other routes, and pose relatively few safety concerns. Compared to other nucleic acid vectors, they are usually devoid of viral or bacterial antigens and can be designed to deliver only the target tumor antigen(s). This is likely to be important when priming a response against weak tumor antigens. DNA vaccines have been more effective in rodents than in larger mammals or humans. However, a large number of methods that might be applied clinically have been shown to ameliorate these vaccines. This includes in vivo electroporation, and/or inclusion of various immunostimulatory molecules, xenoantigens (or their epitopes), antigen-cytokine fusion genes, agents that improve antigen uptake or presentation, and molecules that activate innate immunity mechanisms. In addition, CpG motifs carried by plasmids can overcome the negative effects of regulatory T cells. There have been few studies in humans, but recent clinical trials suggest that plasmid/virus, or plasmid/antigen-adjuvant, prime-boost strategies generate strong immune responses, and confirm the usefulness of plasmid-based vaccination. PMID- 15538730 TI - Highly efficient redirected anti-tumor activity of human lymphocytes transduced with a completely human chimeric immune receptor. AB - BACKGROUND: Novel antibody-based immunotherapeutic strategies exploit chimeric immune receptors (CIR), expressed on the surface of transduced human peripheral blood mononuclear cells (PBMC), to redirect potent non-MHC-dependent cytotoxicity to tumor cells expressing a tumor-associated antigen. However, clinical application of the strategy has been hampered by the potential side effects associated with immunogenicity and by low transduction efficiency. METHODS: A fully human CIR was constructed that triggers immune activation through the zeta chain of CD3 and contains a human single-chain antibody fragment specific for an extracellular epitope of HER2. PBMC were transduced with the CIR using gibbon-ape leukemia virus envelope pseudotyped retroviruses. In vitro cytotoxicity and inhibition assays were carried out using normal and tumor cell lines expressing different levels of HER2. RESULTS: Bulk populations of CIR-transduced PBMC could express high levels of the construct and subcloning ensured stable expression. CIR-mediated killing and growth inhibition of targets expressing high HER2 levels were very efficient at low effector-to-target ratios. Under the same experimental conditions, CIR-mediated activity against normal cells expressing low HER2 levels was marginal. The CIR-mediated recognition of target cells induced the release of soluble factors able to inhibit growth of both HER-positive and HER2-negative bystander tumor cells. CONCLUSIONS: Human CIR-transduced PBMC exert a potent and dose-dependent anti-tumor activity. Target antigen level appeared to be a critical determinant of specificity and delivery of signals leading to redirected effector functions. Soluble factors, released by redirected effectors at the site of antigen-driven activation, mediate potent bystander killing. PMID- 15538732 TI - Improved gene transfer and normalized enzyme levels in primitive hematopoietic progenitors from patients with mucopolysaccharidosis type I using a bioreactor. AB - BACKGROUND: One of the major barriers to the clinical application of hematopoietic stem cell (HSC) gene therapy has been relatively low gene transfer efficiency. Other inadequacies of current transduction protocols are related to their multi-step procedures, e.g., using tissue-culture flasks, roller bottles or gas-permeable bags for clinical application. METHODS: In comparison with a conventional bag transduction protocol, a 'closed' hollow-fiber bioreactor system (HBS) was exploited to culture and transduce human peripheral blood CD34(+) progenitor cells (PBPC(MPS)) from patients with mucopolysaccharidosis type I (MPS I) using an amphotropic retroviral vector based on a murine Moloney leukemia virus LN prototype. Both short-term colony-forming cell (CFC) and long-term culture initiating cell (LTCIC) assays were employed to determine transduction frequency and transgene expression in committed progenitor cells and primitive progenitors with multi-lineage potentials. RESULTS: A novel ultrafiltration transduction method was established to culture and transduce enzyme-deficient PBPC(MPS) over a 5-day period without loss in viability and CD34 identity (n = 5). Significantly higher transduction efficiencies were achieved in primary CFC that derived from the HBS (5.8-14.2%) in comparison with those from gas-permeable bags (undetectable to 1.7%; p < 0.01). Up to 15-fold higher-than-normal enzyme activity was found in selected PBPC(MPS)-LP1CD transductants. Moreover, higher gene transfer (4.4-fold) and expression in very primitive progenitors were observed in products from the HBS compared with bag experiments as indicated by CFC derived from primitive LTCIC. Remarkably, with relatively modest gene transfer levels in LTCIC from HBS experiments, the expression of the IDUA transgene corrected the enzyme-deficiency in 5-week long-term cultures (LTC). CONCLUSIONS: MPS I progenitor cells achieved normalized enzyme levels in LTC after transduction in a HBS system. These studies demonstrate the advantages of a bioreactor-transduction system for viral-mediated stem cell gene transfer. PMID- 15538733 TI - The relationship of VEGF and PGE2 expression to extracellular matrix remodelling of the tenosynovium in the carpal tunnel syndrome. AB - Tenosynovial thickening within the confined space of the carpal tunnel is thought to be the cause of the carpal tunnel syndrome (CTS). However, little is known about the pathological mechanism of tenosynovial thickening. In this study, the role of prostaglandin E(2) (PGE(2)) and vascular endothelial growth factor (VEGF) (two representative molecules that can induce oedema by increasing vascular permeability) was analysed in CTS by using immunohistochemistry and enzyme-linked immunosorptive assay (ELISA). Expression of these molecules was compared with the patients' clinical histories and a temporary increase in production of these molecules was found in cells within the vessels and synovial lining during the intermediate phase of the syndrome when the histology of the tenosynovium changes from oedematous to fibrotic. Statistical analysis clearly demonstrated that there is a close correlation between the expression of PGE(2) and VEGF. Furthermore, immunohistochemical analysis with anti-proliferating cell nuclear antigen (PCNA) revealed that the area with distinct VEGF expression closely matched the area where endothelial cells, vascular smooth muscle cells, and synovial lining cells proliferate. In contrast, despite marked alteration in the extracellular matrix (ECM) component of the tenosynovium, the fibroblasts responsible for most ECM framework production do not proliferate during any phase of CTS. Histological analysis demonstrated that angiogenesis takes place only during the intermediate phase. Since clusters of capillaries and arterioles are often surrounded by type III collagen-rich, disorganized, degenerate connective tissue, which contains fewer fibroblasts than normal, angiogenesis appears to take place as a part of a regenerative reaction that results in fibrosis. These findings strongly indicate that both PGE(2) and VEGF are expressed in the tenosynovium in CTS during the intermediate phase and induce the histological changes seen in the tenosynovium. PMID- 15538734 TI - Granulocyte-derived elastase and gelatinase B are required for dermal-epidermal separation induced by autoantibodies from patients with epidermolysis bullosa acquisita and bullous pemphigoid. AB - Epidermolysis bullosa acquisita (EBA) and bullous pemphigoid (BP) are two clinically and immunologically distinct autoimmune subepidermal blistering skin diseases associated with IgG autoantibodies against the dermal-epidermal junction. BP antibodies are directed against the hemidesmosomal antigens BP180 and BP230, and those in patients with EBA target type VII collagen, a major component of anchoring fibrils. While the pathogenetic mechanisms of subepidermal blistering in BP have been previously studied using a passive transfer mouse model, the effector pathways of blister formation in EBA are largely unknown. Autoantibodies to type VII collagen and BP180 have recently been shown to induce leucocyte-mediated subepidermal cleavage in cryosections of human skin. The aim of the present study was to identify human leucocyte protease(s) instrumental in dermal-epidermal separation induced by autoantibodies to type VII collagen and BP180. When incubated with cryosections of human skin pretreated with IgG from patients with EBA or BP but not from patients with anti-laminin 5 mucous membrane pemphigoid or healthy controls, granulocytes were recruited to the dermal epidermal junction and induced subepidermal splits. A combination of broad-range protease inhibitors as well as inhibitors of serine and matrix metalloproteases completely abolished dermal-epidermal separation induced by EBA or BP autoantibodies. When characterizing the proteases involved more specifically, selective inhibition of human leucocyte elastase or gelatinase B/MMP-9 was also found to result in suppression of blistering. These findings strongly suggest that elastase and gelatinase B are essential for granulocyte-mediated proteolysis resulting in dermal-epidermal separation in EBA and BP patients' skin. PMID- 15538735 TI - Renal ACE2 expression in human kidney disease. AB - Angiotensin-converting enzyme 2 (ACE2) is a recently discovered homologue of angiotensin-converting enzyme (ACE) that is thought to counterbalance ACE. ACE2 cleaves angiotensin I and angiotensin II into the inactive angiotensin 1-9, and the vasodilator and anti-proliferative angiotensin 1-7, respectively. ACE2 is known to be present in human kidney, but no data on renal disease are available to date. Renal biopsies from 58 patients with diverse primary and secondary renal diseases were studied (hypertensive nephropathy n = 5, IgA glomerulopathy n = 8, minimal change nephropathy n = 7, diabetic nephropathy n = 8, focal glomerulosclerosis n = 5, vasculitis n = 7, and membranous glomerulopathy n = 18) in addition to 17 renal transplants and 18 samples from normal renal tissue. Immunohistochemical staining for ACE2 was scored semi-quantitatively. In control kidneys, ACE2 was present in tubular and glomerular epithelium and in vascular smooth muscle cells and the endothelium of interlobular arteries. In all primary and secondary renal diseases, and renal transplants, neo-expression of ACE2 was found in glomerular and peritubular capillary endothelium. There were no differences between the various renal disorders, or between acute and chronic rejection and control transplants. ACE inhibitor treatment did not alter ACE2 expression. In primary and secondary renal disease, and in transplanted kidneys, neo-expression of ACE2 occurs in glomerular and peritubular capillary endothelium. Further studies should elucidate the possible protective mechanisms involved in the de novo expression of ACE2 in renal disease. PMID- 15538736 TI - Dimeric galectin-1 induces IL-10 production in T-lymphocytes: an important tool in the regulation of the immune response. AB - Galectin-1, a beta-galactoside binding protein that can occur as both a monomer and a homodimer, binds to leucocyte membrane antigens such as CD7, CD43, and CD45, and has immune-regulatory functions in several animal models of autoimmune disease. However, its mechanism of action is only partially understood. In this study, a marked increase in IL-10 mRNA and protein levels was demonstrated in non activated and activated CD4(+) and CD8(+) T-cells, following treatment with a high concentration (dimeric form), but not a low concentration (monomeric form), of recombinant galectin-1 protein. IL-10 is known to suppress TH1 type immune responses and upregulation of IL-10 may thus contribute to the immune-regulatory function of galectin-1. Galectin-1 was strongly expressed on the endothelial cells of human kidney allografts, suggesting a role in the regulation of immune responses in transplantation. Administration of high concentrations of galectin-1 may be a useful tool in the treatment of T-cell-mediated diseases. PMID- 15538737 TI - C-C chemokine receptor 2 (CCR2) deficiency improves bleomycin-induced pulmonary fibrosis by attenuation of both macrophage infiltration and production of macrophage-derived matrix metalloproteinases. AB - Macrophage infiltration is implicated in various types of pulmonary fibrosis. One important pathogenetic process associated with pulmonary fibrosis is injury to basement membranes by matrix metalloproteinases (MMPs) that are produced mainly by macrophages. In this study, C-C chemokine receptor 2-deficient (CCR2-/-) mice were used to explore the relationship between macrophage infiltration and MMP activity in the pathogenesis of pulmonary fibrosis, using the bleomycin-induced model of this disease process. CCR2 is the main (if not only) receptor for monocyte chemoattractant protein-1/C-C chemokine ligand 2 (MCP-1/CCL2), which is a critical mediator of macrophage trafficking, and CCR2 -/- mice demonstrate defective macrophage migration. Pulmonary fibrosis was induced in CCR2-/- and wild-type (CCR2+/+) mice by intratracheal instillation of bleomycin. No significant differences in the total protein concentration in bronchoalveolar lavage (BAL) fluid, or in the degree of histological lung inflammation, were observed in the two groups until day 7. Between days 3 and 21, however, BAL fluid from CCR2-/- mice contained fewer macrophages than BAL fluid from CCR2+/+ mice. Gelatin zymography of BAL fluid and in situ zymography revealed reduced gelatinolytic activity in CCR2-/- mice. Immunocytochemical staining showed weaker expression of MMP-2 and MMP-9 in macrophages in BAL fluid from CCR2-/- mice at day 3. Gelatin zymography of protein extracted from alveolar macrophages showed reduced gelatinolytic activity of MMP-2 and MMP-9 in CCR2-/- mice. At days 14 and 21, lung remodelling and the hydroxyproline content of lung tissues were significantly reduced in CCR2-/- mice. These results suggest that the CCL2/CCR2 functional pathway is involved in the pathogenesis of bleomycin-induced pulmonary fibrosis and that CCR2 deficiency may improve the outcome of this disease by regulating macrophage infiltration and macrophage-derived MMP-2 and MMP-9 production. PMID- 15538738 TI - In vivo activation of gelatinase B/MMP-9 by trypsin in acute pancreatitis is a permissive factor in streptozotocin-induced diabetes. AB - Matrix metalloproteinases, in particular gelatinase B/MMP-9, are key mediators in autoimmune diseases like multiple sclerosis and rheumatoid arthritis, but their pathogenic roles in diabetes are not well established. Gelatinase B has previously been shown to be upregulated in pancreas tissue from patients with acute and chronic pancreatitis and was suggested to exacerbate diabetes by cleaving insulin. In this study, the role of gelatinase B in diabetes was investigated using two streptozotocin-induced animal models of type I diabetes. In both a hyperacute and a subacute model, gelatinase B upregulation was found to be associated with disease activity. However, gelatinase B deficiency did not significantly protect against diabetes development, and wild-type and gelatinase B-deficient animals behaved similarly in terms of beta-cell apoptosis or necrosis. The fact that gelatinase B was found almost exclusively as the inactive pro-enzyme in most of the streptozotocin-induced diabetic animals may explain the lack of a gelatinase B effect. On the contrary, gelatinase B was completely activated in a minority (15%) of wild-type animals. This coincided with exocrine pancreatic inflammation, as revealed by the presence of active trypsin. The discovery of in vivo activation of progelatinase B by trypsin in acute pancreatitis is extended in a model of caerulein-induced pancreatitis. In the latter model, trypsinogen activation is systematically achieved and gelatinase B is found in its active form. In conclusion, gelatinase B itself is not a causative factor but, when activated by endogenous trypsin, is a permissive factor for insulin degradation and diabetes. PMID- 15538739 TI - Thymidylate synthase, thymidine phosphorylase, dihydropyrimidine dehydrogenase expression, and histological tumour regression after 5-FU-based neo-adjuvant chemoradiotherapy in rectal cancer. AB - Pre-operative 5-fluorouracil (5-FU)-based chemoradiotherapy in locally advanced rectal cancer (UICC-II/III) may significantly reduce local tumour mass. Response to pre-operative treatment, however, varies significantly. Thymidylate synthase (TS), thymidine phosphorylase (TP), and dihydropyrimidine dehydrogenase (DPD) are thought to be important predictors for the efficiency of 5-FU-based treatment. The aim of this study was to determine the correlation between TS-, TP-, and DPD gene expression and the response to 5-FU-based long-term pre-operative chemoradiotherapy assessed by histopathological tumour regression. Additionally, the predictive value of intra-tumoural TS-, TP-, and DPD-gene expression in pre operative rectal tumour biopsies was assessed by correlation with the histopathological regression grade. Formalin-fixed, paraffin wax-embedded pre operative biopsies (n = 14) and surgical resection specimens (n = 40) from patients with rectal carcinoma (clinical UICC stage II/III) receiving neo adjuvant 5-FU-based chemoradiotherapy were studied for TS-, TP-, and DPD-gene expression by quantitative TaqMan real-time PCR after laser microdissection. Results were compared with standardized histopathological tumour regression analysis. There was a significant association between low TS-gene expression in pre-operative tumour biopsies and tumour response (p = 0.02). TS- and TP-gene expression was significantly lower in resection specimens of responders than of non-responders (p = 0.02) when microdissection was used. Statistical significance was even higher when TS and TP were combined (p = 0.0001). For the DPD gene, no significance was found at all. In conclusion, this study shows that TS gene expression in a pretreatment biopsy predicts the response of local rectal cancer to neo-adjuvant 5-FU-based chemoradiotherapy in a high percentage. Moreover, intra-tumoural TS- and TP-gene expression in surgical rectal specimens after neo adjuvant chemoradiotherapy correlates significantly with histopathological tumour regression when microdissection is applied. PMID- 15538740 TI - Fibroblast growth factor 17 is over-expressed in human prostate cancer. AB - Over-expression of fibroblast growth factor 8 (FGF8) in human prostate cancer is associated with clinically aggressive disease. Among different members of the FGF family, FGF17 and FGF8 share high sequence homology and have similar patterns of expression during embryogenesis. In this study, the clinical significance of FGF17 expression and its in vitro function in prostate cancer cells were tested. Forty resected prostate specimens from patients with benign prostatic hyperplasia (BPH, n = 12) and prostate cancer (CaP, n = 28; Gleason sum scores 3-10) were studied using semi-quantitative RT-PCR. In addition, 85 cases of CaP (Gleason sum scores 5-9) and 20 cases of BPH were examined using immunohistochemistry and findings were correlated with clinical parameters. In vitro experiments using prostate cancer cell lines examined the functional significance of FGF17 in prostate cancer. These studies revealed a significant linear correlation between increasing Gleason sum scores and FGF17 expression using both immunohistochemistry (p < 0.0001, rho = 0.99) and RT-PCR (p = 0.008, rho = 0.99). Immunohistochemistry demonstrated upregulation of FGF17 in CaP compared with BPH (p < 0.0001) and, when comparing high-grade CaP (Gleason sum score 7-10) with BPH, RT-PCR showed a fourfold upregulation of FGF17 mRNA expression (p < 0.0001). Men with tumours displaying high levels of FGF17 expression had a worse outcome on survival analysis (p = 0.044) and a higher risk of progression with metastases (p < 0.0001). Proliferation assays showed low-dose recombinant (r) FGF17 (1 ng/ml) to be a more potent mitogen than rFGF1 and rFGF8 in prostate cancer cell lines (LNCaP, DU145, and PC3M) (p < 0.001). Furthermore, FGF8 was shown to induce expression of FGF17 in these cell lines. These data support a role for FGF17, and a model of co-expression of multiple FGFs, with FGF17 as a potential mediator of FGF8 function, in human prostate carcinogenesis. PMID- 15538741 TI - New and highly efficient method for silkworm transgenesis using Autographa californica nucleopolyhedrovirus and piggyBac transposable elements. AB - We have developed a new method for the transgenesis of the silkworm, Bombyx mori. This method couples the use of recombinant baculoviruses with the use of the piggyBac transposable element. One recombinant AcNPV, designated the helper virus, is designed to express the piggyBac transposase under the control of the Drosophila hsp70 promoter. Another recombinant AcNPV encoded the gene to be incorporated into the silkworm genome, in this case a green fluorescent protein (GFP) gene, under the control of B. mori actin A3 promoter and franked by the piggyBac inverted terminal repeats. Preblastoderm eggs were inoculated with a fine needle coated with a mixture of these two recombinant baculoviruses. Most of the inoculated larvae hatched and a high proportion of the newly hatched G0 larvae expressed the GFP marker. Transgenesis was confirmed by Southern blot analysis of G1 insects, sequencing the insertion site junctions isolated by inverse PCR, and the marker segregated in Mendelian fashion, as evidenced by the appearance of green fluorescence in G2 insects. Thus, transgenic silkworms were easily and efficiently obtained using this new method. PMID- 15538742 TI - Physical and hydrodynamic properties of flocs produced during biological hydrogen production. AB - Dense flocs readily form in continuous culture bioreactors used for hydrogen production, but the fractal and hydrodynamic properties of these flocs have not been previously analyzed. We therefore examined the size distribution, fractal dimension, and hydrodynamic properties of flocs formed in a continuous flow, well mixed reactor treating synthetic wastewater at a fixed condition of a 4.5 h hydraulic detention time (23 degrees C, pH 5.5). The reactor was operated for a total of 3 months at three different organic loading rates (27, 53, and 80 g COD/L-d) with influent glucose concentrations of 5, 10, and 15 g-COD/L. At all three loading rates the removal of glucose was nearly complete (98.6-99.4%) and biomass was produced in proportion to the organic loading rate (0.86 +/- 0.11, 2.40 +/- 0.26, and 4.59 +/- 1.55 g/L of MLVSS in the reactor). Overall conversion efficiencies of glucose to hydrogen, evaluated on the basis of a maximum of 4 mol H2/mol-glucose, increased with organic loading rates in the order 17.7%, 23.1%, and 25.6%. The gas contained 56.1 +/- 4.9% hydrogen, with the balance as carbon dioxide. No methane gas was detected. Under these conditions, flocs were produced with mean sizes that increased with organic loading, in the order 0.12 cm (5 g COD/L), 0.35 cm (10 g-COD/L), and 0.58 cm (15 g-COD/L). As the average floc size increased, the flocs became on average denser and less fractal, with fractal dimensions increasing from 2.11 +/- 0.17 to 2.48 +/- 0.13. Floc porosities ranged from 0.75-0.96, and resulted in aggregate densities that allowed little intra aggregate flow through the floc. As a result, average settling velocities were not appreciably larger than those predicted by Stokes' law for spherical, impermeable flocs. Our results demonstrate that dense, relatively impermeable flocs are produced in biohydrogen reactors that have settling properties in reasonable agreement with Stokes' law. PMID- 15538743 TI - Association of CYP17, GSTP1, and PON1 polymorphisms with the risk of prostate cancer. AB - BACKGROUND: Dietary factors, life-style as well as environmental conditions may contribute to the risk of prostate tumor together with genetic susceptibility, that may be an important factor in determining who is more likely to develop prostate malignancy. We have undertaken a case-control study in order to elucidate the association between polymorphisms in some metabolizing genes with the risk of prostate cancer (PCa). METHODS: Polymorphisms of three xenobiotic genes (CYP17, GSTP1, PON1) were characterized in 384 patients with untreated PCa and 360 age-matched control patients with benign prostatic hyperplasia (BPH). All polymorphisms were investigated by PCR/RFLP methods using DNA from lymphocytes. RESULTS: We found that men with the CYP17/A1A1-A1A2 genotypes, GSTP1/IleVal genotype, PON192/QR and PON55/LM-MM genotypes had a significantly higher risk of PCa compared with the others genotypes. CONCLUSIONS: The three polymorphisms appear to be common genetic traits that are associated with an increased risk for PCa: the analysis of them all in each single case may be a predictable factor, particularly among groups exposed to PCa-related carcinogens. PMID- 15538745 TI - 5alpha-dihydrotestosterone inhibits 1alpha,25-dihydroxyvitamin D3-induced expression of CYP24 in human prostate cancer cells. AB - BACKGROUND: A cross-talk between 1alpha,25-dihydroxyvitamin D(3) [1alpha,25 (OH)(2)D(3)] and 5alpha-dihydrotestosterone (DHT) in the growth inhibition has been demonstrated, but the mechanism is unknown. METHODS: The expression of 25 hydroxyvitamin D(3) 24-hydroxylase (24-hydroxylase) was measured using a real time quantitative RT-PCR assay and the catabolism of 1alpha,25-(OH)(2)D(3) was measured using a radioreceptor assay. RESULTS: Real-time RT-PCR showed that DHT at 1-100 nM significantly inhibited 1alpha,25-(OH)(2)D(3)-induced expression of 24-hydroxylase in LNCaP cells. Furthermore, the catabolism of 1alpha,25 (OH)(2)D(3) was decreased by 10 nM DHT. An androgen receptor (AR) antagonist, Casodex antagonized the DHT effect, whereas an AR agonist (due to the mutant AR in LNCaP cells) hydroxyflutamide did not. CONCLUSIONS: We demonstrated, for the first time, that DHT reduces the ability of 1alpha,25-(OH)(2)D(3) to induce 24 hydroxylase expression. Our results not only support the earlier finding of a cross-talk between androgen and vitamin D in human prostate cancer cells but also provide a possible mechanism how androgen and vitamin D signaling pathways may interact. PMID- 15538746 TI - Differential alterations in 5alpha-reductase type 1 and type 2 levels during development and progression of prostate cancer. AB - BACKGROUND: In the prostate, conversion of testosterone to dihydrotestosterone (DHT), by the enzymes 5alpha-reductase types 1 and 2 (5alphaR1, 5alphaR2) is required for normal growth and probably also for development of prostate cancer (PCa). Finasteride, a 5alphaR2 inhibitor, was shown to reduce the prevalence of PCa in the Prostate Cancer Prevention Trial. However, inhibition of both 5alphaR isoenzymes causes a greater decrease in serum DHT. The aim of this study was to assess differential expression of these enzymes at various stages of PCa development. METHODS: Immunostaining for 5alphaR1 and 5alphaR2, using specific, well-validated antibodies, was evaluated in 26 benign prostatic hyperplasia (BPH) (16 for 5alphaR2), 53 primary PCa (21 for 5alphaR2), 18 prostatic intraepithelial neoplasia (PIN), 12 primary PCa treated with neoadjuvant androgen ablation, 15 locally recurrent PCa specimens, and 18 PCa metastases. RESULTS: The mean area of moderate plus high intensity staining for 5alphaR1 increased from 4.8 +/- 2.8% of total epithelial area in BPH, to 18.9 +/- 5.7% in PIN, 17.0 +/- 3.2% in primary cancer, 38.0 +/- 7.3% in recurrent cancer, and 55.8 +/- 8.5% in PCa metastases. The mean staining area for 5alphaR2 decreased from 58.8 +/- 7.2% in BPH, to 21.1 +/- 5.5% in PIN and 34.8 +/- 6.7% in primary PCa. Staining for 5alphaR2 was increased in recurrent cancer and PCa metastases compared to primary PCa, at 58.7 +/- 5.2% and 69.2 +/- 8.7%, respectively. CONCLUSIONS: 5alphaR1 immunostaining is increased and 5alphaR2 immunostaining is decreased during development of PCa. In addition, there is increased expression of both 5alphaR isozymes in recurrent and metastatic cancers, suggesting that both isozymes may be important in the development and progression of PCa. PMID- 15538747 TI - Phosphoproteomics by mass spectrometry and classical protein chemistry approaches. AB - The general fields of biological sciences have seen phenomenal transformations in the past two decades at the level of data acquisition, understanding biological processes, and technological developments. Those advances have been made partly because of the advent of molecular biology techniques (which led to genomics) coupled to the advances made in mass spectrometry (MS) to provide the current capabilities and developments in proteomics. However, our current knowledge that approximately 30,000 human genes may code for up to 1 million or more proteins disengage the interface between the genome sequence database algorithms and MS to generate a major interest in independent de novo MS/MS sequence determination. Significant progress has been made in this area through procedures to covalently modify peptide N- and C-terminal amino-acids by sulfonation and guanidination to permit rapid de novo sequence determination by MS/MS analysis. A number of strategies that have been developed to perform qualitative and quantitative proteomics range from 2D-gel electrophoresis, affinity tag reagents, and stable isotope labeling. Those procedures, combined with MS/MS peptide sequence analysis at the subpicomole level, permit the rapid and effective identification and quantification of a large number of proteins within a given biological sample. The identification of proteins per se, however, is not always sufficient to interpret biological function because many of the naturally occurring proteins are post-translationally modified. One such modification is protein phosphorylation, which regulates a large array of cellular biochemical pathways of the biological system. Traditionally, the study of phosphoprotein structure function relationships involved classical protein chemistry approaches that required protein purification, peptide mapping, and the identification of the phosphorylated peptide regions and sites by N-terminal sequence analysis. Recent advances made in mass spectrometry have clearly revolutionized the studies of phosphoprotein biochemistry, and include the development of specific strategies to preferentially enrich phosphoproteins by covalent-modifications that incorporate affinity tags that use the physicochemical properties of phosphoaminoacids. The phosphoserine/phosphothreonine-containing proteins/peptides are derivatized under base-catalyzed conditions by thiol agents; mono- and di-thiol reagents both have been used in such studies. The thiol agent may have: (i) an affinity tag for protein enrichment; (ii) stable isotopic variants for relative quantitation; or (iii) a combination of the moieties in (i) and (ii). These strategies and techniques, together with others, are reviewed, including their practical application to the study of phosphoprotein biochemistry and structure-function. The consensus of how classical protein chemistry and current MS technology overlap into special case of proteomics, namely "phosphoproteomics," will be discussed. PMID- 15538748 TI - Identification of hypoxia-inducible factor-1alpha (HIF-1alpha) polymorphism as a mutation in prostate cancer that prevents normoxia-induced degradation. AB - BACKGROUND: Hypoxia-inducible factor-1alpha (HIF-1alpha) regulates cellular responses to hypoxia and is rapidly degraded under normoxia through von Hippel Lindau (VHL) mediated ubiquitination. Although HIF-1alpha stabilization appears to be the molecular basis for VHL-associated cancers, stabilizing mutations in HIF-1alpha have not been reported. METHODS: A series of 15 metastatic androgen independent prostate cancers were examined for mutations in the oxygen-dependent domain (ODD) of HIF-1alpha by PCR amplification and DNA sequencing. RESULTS: A somatic proline to serine mutation in codon 582 (P582S) was identified in one sample. Transfection studies with a HIF-1alpha regulated reporter gene showed increased transcriptional activity that correlated with higher mutant HIF-1alpha protein expression. Increased expression of the P582S mutant induced by iron chelation, which blocks proline hydroxylation of wild-type HIF-1alpha, was markedly attenuated. The mutant also showed increased stability under normoxic versus hypoxic conditions. CONCLUSION: The P582S HIF-1alpha is a stable variant and HIF-1alpha mutation is a mechanism for enhancing HIF-1alpha activity in human cancer. The recent identification of the identical P582S HIF-1alpha as a polymorphism suggests that this variant may increase tumor susceptibility or cause more aggressive biological behavior. PMID- 15538749 TI - Glial transcytosis of a photoreceptor-secreted signaling protein, retinoschisin. AB - In vitro studies have clearly shown that signaling/guidance proteins can diffuse to their targets. However, it is unclear whether they can travel by diffusion in vivo, or if they are distributed in the tissue by an active mechanism. Retinoschisin, a signaling molecule related to neuropilins, is synthesized and secreted by photoreceptor cells in the outer retina; then it interacts with inner retinal cells contributing to synaptic organization and optic nerve fiber integrity. We developed an assay to examine how retinoschisin, which is secreted a distance away, reaches its inner retinal targets. We found that retinoschisin is preferentially taken up and carried into the inner retina from the retinal outer border (the photoreceptor side) by Muller cells (the main glial cells of the vertebrate retina). This transcytosis is disrupted by DL-alpha-aminoadipic acid, a Muller cell/glia-specific toxin. Our results suggest that glial uptake/transcytosis can provide an effective and precise alternative for distributing signaling molecules in the nervous system. PMID- 15538750 TI - Mutations and elevated transcriptional activity of conductin (AXIN2) in hepatoblastomas. AB - Hepatoblastoma (HB) is the most frequent malignant liver tumour of childhood. Most HBs develop sporadically but their incidence is highly elevated in patients with familial adenomatous polyposis coli (FAP). These patients carry germline mutations in the adenomatous polyposis coli (APC) tumour suppressor gene. APC forms a multi-protein complex involved in the WNT signalling pathway that controls the stability of beta-catenin, the central effector in this cascade. Whereas APC mutations are rare in sporadic HBs, a high frequency of beta-catenin mutations leading to overactivation of WNT signalling was previously found in these tumours. This pathway is negatively controlled by conductin (axin2), representing a further partner in this signalling complex. To investigate whether alterations in conductin may also be involved in the pathogenesis of sporadic HBs, 37 HBs and five HB cell lines were screened for mutations using single strand conformation polymorphism (SSCP) analysis, reverse transcription polymerase chain reaction (RT-PCR), and direct sequencing. In two cases, larger deletions (52 and 1624 bp) leading to frameshifts were found. In addition, one HB carried a somatic point mutation. Expression analysis by competitive RT-PCR in HBs revealed up-regulation of conductin mRNA compared with adjacent liver samples. This mRNA overexpression resulted in increased conductin protein levels demonstrated by western blot analysis. Tumours with activating beta-catenin mutations revealed higher levels of conductin mRNA transcripts. This finding indicates that conductin is a direct target gene of WNT signalling in HBs, as has been demonstrated in other tissues. In summary, conductin mutations may represent an alternative mechanism leading to activation of WNT signalling in HBs. The overexpression of conductin mRNA in HBs reflects activation of the WNT pathway because conductin represents a target gene of WNT signalling in liver tissue. PMID- 15538751 TI - Human embryonic stem cells differentiate into oligodendrocytes in high purity and myelinate after spinal cord transplantation. AB - Human embryonic stem cells (hESCs) demonstrate remarkable proliferative and developmental capacity. Clinical interest arises from their ability to provide an apparently unlimited cell supply for transplantation, and from the hope that they can be directed to desirable phenotypes in high purity. Here we present for the first time a method for obtaining oligodendrocytes and their progenitors in high yield from hESCs. We expanded hESCs, promoted their differentiation into oligodendroglial progenitors, amplified those progenitors, and then promoted oligodendroglial differentiation using positive selection and mechanical enrichment. Transplantation into the shiverer model of dysmyelination resulted in integration, differentiation into oligodendrocytes, and compact myelin formation, demonstrating that these cells display a functional phenotype. This differentiation protocol provides a means of generating human oligodendroglial lineage cells in high purity, for use in studies of lineage development, screening assays of oligodendroglial-specific compounds, and treating neurodegenerative diseases and traumatic injuries to the adult CNS. PMID- 15538752 TI - Caspr reveals an aggregation of nodes and flanking node free zones at the rat trigeminal sensory root and dorsal root entry zones. AB - The sensory root entry zone demarcates the transition from the peripheral nervous system (PNS) to the central nervous system (CNS). In this study, we describe the organization of nodes of Ranvier at the trigeminal sensory and dorsal root entry zones of the rat. Caspr immunoreactivity (IR) was used to identify the paranodal region of nodes of Ranvier, while L-MAG-IR was used to identify CNS oligodendrocytes. Immunofluorescence confocal microscopy revealed a dense aggregation of nodes precisely at the PNS to CNS transition with prominent node depleted zones on either side, while L-MAG-IR was confined to ensheathing fibers on the central side of nodes located in this dense band and identified these as transitional nodes. Morphometric analysis of the PNS and CNS sides of the trigeminal and the PNS side of the dorsal root entry zones confirmed the presence of virtually node-free domains flanking the transitional zone. Further, the reappearance of nodes on the far side of the node-free zones strongly correlated with nodal diameter, with small nodes reappearing first. These findings suggest that the PNS/CNS transition may represent the initial site of myelination of the primary afferent axon within this area. PMID- 15538753 TI - Differential activation of astrocytes by innate and adaptive immune stimuli. AB - The immunologic privilege of the central nervous system (CNS) makes it crucial that CNS resident cells be capable of responding rapidly to infection. Astrocytes have been reported to express Toll-like receptors (TLRs), hallmark pattern recognition receptors of the innate immune system, and respond to their ligation with cytokine production. Astrocytes have also been reported to respond to cytokines of the adaptive immune system with the induction of antigen presentation functions. Here we have compared the ability of TLR stimuli and the adaptive immune cytokines interferon-gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) to induce a variety of immunologic functions of astrocytes. We show that innate signals LPS- and poly I:C lead to stronger upregulation of TLRs and production of the cytokines IL-6 and TNF-alpha as well as innate immune effector molecules IFN-alpha4, IFN-beta, and iNOS compared with cytokine stimulated astrocytes. Both innate stimulation and adaptive stimulation induce similar expression of the chemokines CCL2, CCL3, and CCL5, as well as similar enhancement of adhesion molecule ICAM-1 and VCAM-1 expression by astrocytes. Stimulation with adaptive immune cytokines, however, was unique in its ability to induce upregulation of MHC II and the functional ability of astrocytes to activate CD4(+) T cells. These results indicate potentially important and changing roles for astrocytes during the progression of CNS infection. PMID- 15538754 TI - Mannose receptor expression specifically reveals perivascular macrophages in normal, injured, and diseased mouse brain. AB - Perivascular macrophages are believed to have a significant role in inflammation in the central nervous system (CNS). They express a number of different receptors that point toward functions in both innate immunity, through pathogen-associated molecular pattern recognition, phagocytosis, and cytokine responsiveness, and acquired immunity, through antigen presentation and co-stimulation. We are interested in the receptors that are differentially expressed by perivascular macrophages and microglia in both the normal CNS as well as in neuroinflammation and neurodegeneration. In this article we report the use of a well-characterized monoclonal antibody, 5D3, to localize the expression of the mannose receptor to perivascular macrophages in the normal CNS and in various models of brain pathology. Mannose receptor expression was limited to perivascular, meningeal, and choroid plexus macrophages in normal, inflamed, injured, and diseased CNS. In particular, activated microglia and invading hematogenous leukocytes were mannose receptor negative while expressing the F4/80 antigen, macrosialin (CD68), FcRII (CD32), scavenger receptor (CD204), and CR3 (CD11b/CD18). Since the perivascular macrophages expressing the mannose receptor are known to be the only constitutively phagocytic cells in the normal CNS, we injected clodronate-loaded liposomes intracerebroventricularly in control mice to deplete these cells. In these mice, there was no detectable mannose receptor expression in perivascular spaces after immunocytochemistry with the 5D3 monoclonal antibody. This finding underlines the value of the monoclonal antibody 5D3 as a tool to study murine perivascular macrophages selectively. Mannose receptor expression by macrophages located at blood-brain (perivascular), brain-cerebrospinal fluid (CSF) (meningeal), and CSF-blood (choroid plexus) interfaces supports a functional role of these cells in responding to external stimuli such as infection. PMID- 15538755 TI - The early transcription factor GATA-2 is expressed in classical Hodgkin's lymphoma. AB - Hodgkin/Reed-Sternberg (HRS) cells of classical Hodgkin's lymphoma (cHL) are thought to be derived from germinal centre B-cells in almost all cases. However, expression profiling has revealed that HRS cells do not show a germinal centre B cell-like phenotype. Although the nature of this aberrant phenotype and the underlying molecular mechanisms remain largely unknown, it has been reported that the activity of NOTCH1 plays an important role in the growth and survival of HRS cells. In some leukaemic cell lines, the effect of Notch signalling is mediated by the early transcription factor GATA-2. This and the fact that HRS cells lack expression of PU.1, which can repress Gata-2, led to an investigation of GATA-2 expression in HRS cells. GATA-2 expression was found in all the cHL-derived cell lines studied, but not in a Burkitt lymphoma-derived cell line. In addition, 50% of biopsies from patients with cHL contained GATA-2-expressing HRS cells. In contrast, neither normal germinal centre B-cells nor malignant cells of nodular lymphocyte-predominant Hodgkin's lymphoma, Burkitt lymphoma or diffuse large B cell lymphoma expressed GATA-2. Thus, GATA-2 expression was found specifically in HRS cells of cHL, suggesting that GATA-2 is important in establishing the abnormal B-cell phenotype of HRS cells. PMID- 15538756 TI - Reversed operation of glutamate transporter GLT-1 is crucial to the development of preconditioning-induced ischemic tolerance of neurons in neuron/astrocyte co cultures. AB - Sublethal ischemia leads to increased tolerance against subsequent prolonged cerebral ischemia in vivo. In the present study, we investigated the roles of the astrocytic glutamate (Glu) transporter GLT-1 in preconditioning (PC)-induced neuronal ischemic tolerance in cortical neuron/astrocyte co-cultures. Ischemia in vitro was simulated by subjecting cultures to both oxygen and glucose deprivation (OGD). A sublethal OGD (PC) increased the survival rate of neurons significantly when cultures were exposed to a lethal OGD 24 h later. The extracellular concentration of Glu increased significantly during PC, and treatment with an inhibitor of N-methyl-D-actetate (NMDA) receptors significantly reversed the PC induced ischemic tolerance of neurons, suggesting that the increase in extracellular concentration of Glu during PC was critical to the development of PC-induced neuronal ischemic tolerance via the activation of NMDA receptors. Treatment with a GLT-1 blocker during PC suppressed this increase in Glu significantly, and antagonized the PC-induced neuronal ischemic tolerance. This study suggested that the reversed operation of GLT-1 was crucial to the development of neuronal ischemic tolerance. PMID- 15538757 TI - Integrated nanoparticle-biomolecule hybrid systems: synthesis, properties, and applications. AB - Nanomaterials, such as metal or semiconductor nanoparticles and nanorods, exhibit similar dimensions to those of biomolecules, such as proteins (enzymes, antigens, antibodies) or DNA. The integration of nanoparticles, which exhibit unique electronic, photonic, and catalytic properties, with biomaterials, which display unique recognition, catalytic, and inhibition properties, yields novel hybrid nanobiomaterials of synergetic properties and functions. This review describes recent advances in the synthesis of biomolecule-nanoparticle/nanorod hybrid systems and the application of such assemblies in the generation of 2D and 3D ordered structures in solutions and on surfaces. Particular emphasis is directed to the use of biomolecule-nanoparticle (metallic or semiconductive) assemblies for bioanalytical applications and for the fabrication of bioelectronic devices. PMID- 15538758 TI - An octahedral rhodium cluster with six phosphine and 12 hydride ligands and 10 too few electrons. PMID- 15538759 TI - Evaluation of cytocompatibility and bending modulus of nanoceramic/polymer composites. AB - In an attempt to simulate the microstructure and mechanical properties of natural bone, novel nanoceramic/polymer composite formulations were fabricated and characterized with respect to their cytocompatibility and mechanical properties. The bending moduli of nanocomposite samples of either poly(L-lactic acid) (PLA) or poly(methyl methacrylate) (PMMA) with 30, 40, and 50 wt % of nanophase (<100 nm) alumina, hydroxyapatite, or titania loadings were significantly (p < 0.05) greater than those of pertinent composite formulations with conventional, coarser grained ceramics. The nanocomposite bending moduli were 1-2 orders of magnitude larger than those of the homogeneous, respective polymer. For example, compared with 0.06 GPa for the 100% PLA, the bending modulus of 50/50 nanophase alumina/PLA composites was 3.5 GPa. Osteoblast adhesion on the surfaces of the nanophase alumina/PLA composites increased as a function of the nanophase ceramic content. Most importantly, osteoblast adhesion on the 50/50 nanophase alumina/PLA substrates was similar to that observed on the 100% nanophase ceramic substrates. Similar trends of osteoblast adhesion were observed on the surfaces of the nanophase titania/polymer and nanophase hydroxyapaptite/polymer composites that were tested. In contrast, fibroblast adhesion on the nanophase composites was either similar or lower than that observed on the conventional composites with either PLA or PMMA and minimum on all tested neat nanophase substrates. The calcium content in the extracellular matrix of cultured osteoblasts was also enhanced on the nanoceramic/PLA composite substrates tested as a function of the nanophase ceramic loading and duration of cell culture. The results of the present in vitro study provide evidence that nanoceramic/polymer composite formulations are promising alternatives to conventional materials because they can potentially be designed to match the chemical, structural, and mechanical properties of bone tissue in order to overcome the limitations of the biomaterials currently used as bone prostheses. PMID- 15538760 TI - Isolation and identification of fungi from vaginal flora in three species of captive Leontopithecus. AB - The ability to reproduce in captivity is an essential component of lion tamarin (Leontopithecus) conservation programs. However, infections such as vaginitis, cervicitis, and endometritis are important diseases that may influence the reproduction of these animals. Therefore, it is important to detect continuous or occasional vaginal microbial populations, and to understand their potential role as an endogenous source of infection [Collins, 1964; Blue, 1983; Pugh et al., 1986]. Vaginal swabs were collected from 25 female tamarins of the three currently available species (L. rosalia, L. chrysopygus, and L. chrysomelas) at the Center of Primatology in Rio de Janeiro, Brazil. The swabs were processed according to standard mycological protocols, and isolates were biochemically characterized. Fungal isolates were recovered from 16 animals (64.0%). The results showed that 70.6% of the isolated microorganisms consisted of yeast, including three species of Candida (mainly C. glabrata). We suggest that this species is a resident member of the normal vaginal flora in Leontopithecus. Filamentous fungi (mainly from Trichosporon, Aspergillus, and Penicilliumgenera) constituted 29.4% of the isolates, and were considered to be transitory contaminants of the genital area. We suggest that colonization of the vaginal environment is related to the endocrine pattern associated with the reproductive status of these animals, but not to parity. PMID- 15538761 TI - Agonistic behavior and dominance relationships in female Phayre's leaf monkeys -- preliminary results. AB - Socioecological theory suggests a link between the strength of competition for food/safety, rates of agonism, structure of dominance hierarchies, and dispersal among group-living females. This study presents preliminary data on agonistic behavior and dominance relationships for female Phayre's leaf monkeys (Trachypithecus phayrei), a species in which females routinely disperse. Behavioral observations were conducted on two groups (four adult females, and five adult females plus two juvenile females, respectively) at Phu Khieo Wildlife Sanctuary, northeast Thailand. Rates of agonistic behavior were analyzed from focal continuous recordings, while dominance hierarchies were constructed from all agonistic behaviors (focal and ad libitum sampling). Overall, female-female agonistic behaviors (aggression, submission, and displacements) occurred at a rate of < 0.25 interactions per hour. Agonistic interactions involving food occurred more frequently than expected based on feeding time. Females in both groups exhibited linear dominance hierarchies with some reversals, and possibly an age-inversed hierarchical structure in the larger group. The results fit well with previous results for colobine monkeys regarding frequency of interactions, displacements predominating agonistic behavior, and the possibility of an age inversed hierarchy. The results contradict the suggested link between linearity of hierarchies and female philopatry. Future studies should consider the notion that female dispersal may coexist with linear dominance hierarchies. PMID- 15538762 TI - Unique proximal tibial morphology in strepsirrhine primates. AB - Although the morphology of the tibial plateau in primates has received very little attention in the literature, it does exhibit features of phylogenetic and functional interest. This paper describes the morphology of the tibial plateau (particularly the intercondylar region) in extant and fossil primates, and in three mammalian outgroups: the pen-tailed tree shrew (Ptilocercus), tree shrew (Tupaia), and flying lemur or dermopteran (Cynocephalus). Extant and fossil strepsirrhine primates exhibit an eminence with a single spine, which contrasts with the intercondylar morphology of haplorhine primates. Most extant platyrrhines, all catarrhine primates (including humans), and some fossil haplorhines possess an eminence with two spines (medial and lateral) connected by a ridge of bone that intersects the intercondylar groove. Tarsius and callitrichines possess an eminence with a reduced medial spine that superficially resembles that of strepsirrhine primates. Dermopterans also exhibit a morphology similar to that of strepsirrhines. In Scandentia, the intercondylar morphology of Tupaia is similar to that of rodents, whereas Ptilocercus resembles tarsiers and callitrichines. We hypothesize that proximal tibiae with either a single spine or reduced medial spine morphology facilitate a greater degree of knee rotation about the eminence relative to the double-spine condition, and are likely associated with more frequent adoption of vertical body positions. In contrast, a double-spine eminence limits knee rotation and is probably associated with greater use of horizontal supports. Although the polarity is complicated by the unknown phylogenetic status of likely sister taxa, it seems most probable that the single-spine morphology is a derived feature of strepsirrhines. PMID- 15538763 TI - Characterizing gastrointestinal transit time in four lemur species using barium impregnated polyethylene spheres (BIPS). AB - Differences in dietary profiles and gastrointestinal (GI) morphologies observed across lemur species suggest that there may be variation in patterns of digesta flow through the GI tract related to the method of digesta processing. Using radio-opaque barium-impregnated polyethylene spheres (BIPS), we characterized such patterns in four lemur species: Varecia variegata (VV), Eulemur fulvus (EF), Propithecus verreauxi (PV), and Hapalemur griseus (HG) (n = 2 per species). After an initial radiograph was taken under light sedation, the animals were fed the BIPS together with a small meal. A combination of 30 small (1.5 mm) and 10 large (5 mm) BIPS was administered. Radiographs were then taken on a species-dependent basis up to 48 hr post-dosage. For small BIPS, the gastric transit time (GTT; time of first exit of BIPS from stomach) was 0.25-2 hr for VV, EF, and HG, and approximately 10 hr for PV. The oro-rectal transit time (ORTT; time of first appearance in the rectum) was < 2 hr for VV and EF, and 24.0 hr for PV and HG. The intestinal transit time (ITT, measured as ORTT - GTT) was < 1.5 hr for VV and EF, and approximately 14 hr and 22 hr for PV and HG, respectively. These data suggest that the GTT of digesta as measured with BIPS was rapid for VV, EF, and HG. For VV and EF, the ORTT and ITT were also rapid, while for HG they were much slower. PV was characterized by delayed GTT, and a more rapid ITT compared to HG. Thus, patterns of flow for PV and HG, despite similar ORTT, differed in that HG emptied BIPS more rapidly and ITT was slower. The flow of BIPS did not differ for VV and EF. These data reveal new information in addition to the total tract transit time, and complement existing knowledge regarding anatomy and diet. PMID- 15538764 TI - Fast and reliable sexing of prosimian and human DNA. AB - Molecular sexing of mammals is normally done by PCR amplification of Y chromosomal fragments, or coamplification of homologous fragments from both sex chromosomes. Existing primers are often unreliable for distantly related species due to mutations in primer regions. Currently there are no published primers for the sexing of prosimian DNA. We show that an existing method (using the zinc finger protein) based on a size difference between the X and Y homologs does not work in prosimians. Multiple alignments of distantly related mammalian species from Genbank and genome databases enabled us to identify conserved regions in the amelogenin gene. Using these conserved regions, we can target species that have no sequence information. We designed a single, conserved primer pair that is useful for fast and reliable molecular sexing of prosimian primates. A single PCR yields two fragments in males and only one in females, which are easily separated with the use of agarose gels. Amplification of separable fragments was successful in seven species of lemurs, as well as humans. PMID- 15538765 TI - Mitochondrial DNA genealogy of chimpanzees in the Nimba mountains and Bossou, West Africa. AB - The chimpanzee populations of the Bossou and Nimba regions in West Africa were genetically surveyed to 1) reveal the genetic relationship between the Bossou and Nimba populations, and 2) elucidate the evolutionary relationship between the Bossou-Nimba and other West African populations. The chimpanzee group at Bossou is characterized by its small population size, no evidence of contact with neighboring populations, and no female immigration. It is believed that most females and adolescent males emigrate from this population. To reveal the genetic signature of these characteristics, we examined the genetic diversity of Bossou and two neighboring populations (Seringbara and Yeale) in the Nimba Mountains by sequencing approximately 605 bp of the mitochondrial DNA (mtDNA) control region. A total of 20 distinct mtDNA variants were observed from 56 sequences of noninvasively collected, anonymous samples. Nucleotide diversity in the Nimba Mountain populations was 0.03-0.04, and did not differ significantly from that in the Bossou population. Very few mitochondrial variants are shared among the sites sampled, which suggests that there is little gene flow involving mtDNA. Nevertheless, no clear population structures were revealed in either population. A comparison with published sequences from West African chimpanzees (Pan troglodytes verus) indicates that the variants observed in the Bossou and Nimba regions are scattered throughout the subspecies, rather than clustered according to geographic region. This suggests that the Bossou-Nimba populations derived only recently from the common ancestral population of the West African chimpanzees, and did not pass through a bottleneck. PMID- 15538766 TI - Nonlinear acoustics in pant hoots of common chimpanzees (Pan troglodytes): frequency jumps, subharmonics, biphonation, and deterministic chaos. AB - The pant hoot calls produced by common chimpanzees (Pan troglodytes) are multi call vocalizations that have figured prominently in investigations of acoustic communication in this species. Although pant hoots are predominantly harmonically structured, they can exhibit an acoustic complexity that has recently been linked to nonlinearity in the vocal-fold dynamics underlying typical mammalian sound production. We examined the occurrence of these sorts of nonlinear phenomena in pant hoot vocalizations, contrasting quieter and lower-pitched "introduction" components with loud and high-pitched "climax" calls in the same bouts. Spectrographic evidence revealed four kinds of nonlinear phenomena, including discrete frequency jumps, subharmonics, biphonation, and deterministic chaos. While these events were virtually never observed during the introduction, they occurred in more than half of the climax calls. Biphonation was by far the most common phenomenon, followed by subharmonics, chaos, and frequency jumps. Individual callers varied in the degree to which their climax calls exhibited nonlinear phenomena, but were consistent in showing more biphonation than other forms. These outcomes show that nonlinear phenomena are routinely present in chimpanzee pant hoots, and help lay the foundation for investigating the function of such events. PMID- 15538767 TI - Use of total dietary fiber across four lemur species (Propithecus verreauxi coquereli, Hapalemur griseus griseus, Varecia variegata, and Eulemur fulvus): does fiber type affect digestive efficiency? AB - In vivo digestibility and transit of two experimental diets were compared across four lemur species for which gastrointestinal morphology and preliminary data on physiology differ:Varecia variegata (VV), Eulemur fulvus (EF), Propithecus verreauxi (PV), and Hapalemur griseus (HG). Since free-ranging groups consume varied amounts of slowly fermentable insoluble fiber (IF) and rapidly fermentable soluble fiber (SF), differences in digestibility may be related to variation in the fiber types consumed. To investigate this, two diets were designed to provide 28% of dry matter (DM) as total dietary fiber (TDF). The ratio of IF/SF (g/g) differed across the diets (12.15:1 for the IF diet, and 3.76:1 for the IF/SF diet). The DM digestibility (DMD) of both diets differed across species: DMD was lower for EF and VV (approximately 56-58%), and higher for PV (72%) and HG (76%). The fiber digestibility results were as follows: TDF digestibility was similar for VV and EF (23% and 28%), higher for PV (56%), and highest for HG (66%). IF digestibility was lower for VV and EF (20% and 28%), and higher for PV and HG (53% and 62%). The transit times (TTs) of the two markers Cr and Co were similar (approximately 3.5 hr for VV and EF, 25 hr for PV, and 30 hr for HG). The mean retention times (MRTs) showed the same trend. The results from these captive groups suggest there are large differences in digestive efficiency that are likely related to the varied fiber composition of the free-ranging diet, and the amount of time the digesta are retained in the gut. PMID- 15538768 TI - From nest to nest--influence of ecology and reproduction on the active period of adult Gombe chimpanzees. AB - The time spent between sleeping periods, which is called the active period, has to accommodate all essential activities, including feeding, resting, social behavior, and reproduction. To minimize costs in terms of, e.g., predation risk, suboptimal foraging, or sleep deficiency, the active period of diurnal animals should be less than or equal to the daylight period. Thus, the active period of an animal should be shaped by local environmental conditions as well as by metabolic and reproductive demands. Chimpanzees, which exhibit reduced predator pressure and a flexible fission-fusion society, were chosen as a model to explore these links. We investigated the influence of sex, female reproductive status, dominance rank, and season on the duration of the active period of adult chimpanzees at Gombe National Park, Tanzania (1975-1992). Sexually nonreceptive females had shorter active periods compared to males, while receptive females had even longer active periods than males. Dominance rank did not influence the duration of the active period of nonreceptive females, but high- and middle ranking males had shorter active periods compared to low-ranking males. Nonreceptive females exhibited longer active periods during the dry season than in the wet season. No seasonal effect was discovered for males, perhaps because they already had long active periods in the wet season. Nonreceptive females seem to be able to accommodate all essential activities in the daylight period available, probably because they live less socially than males. Thus, the active period does not reflect differences in female competitive abilities, but does reflect such differences in males. The duration of the active period appears to be a simple, reliable tool for exploring basic responses and constraints in animal societies. PMID- 15538769 TI - Parallelization of MRCI based on hole-particle symmetry. AB - The parallel implementation of multireference configuration interaction program based on the hole-particle symmetry is described. The platform to implement the parallelization is an Intel-Architectural cluster consisting of 12 nodes, each of which is equipped with two 2.4-G XEON processors, 3-GB memory, and 36-GB disk, and are connected by a Gigabit Ethernet Switch. The dependence of speedup on molecular symmetries and task granularities is discussed. Test calculations show that the scaling with the number of nodes is about 1.9 (for C1 and Cs), 1.65 (for C2v), and 1.55 (for D2h) when the number of nodes is doubled. The largest calculation performed on this cluster involves 5.6 x 10(8) CSFs. PMID- 15538770 TI - An efficient molecular docking using conformational space annealing. AB - Molecular docking falls into the general category of global optimization problems because its main purpose is to find the most stable complex consisting of a receptor and its ligand. Conformational space annealing (CSA), a powerful global optimization method, is incorporated with the Tinker molecular modeling package to perform molecular docking simulations of six receptor-ligand complexes (3PTB, 1ULB, 2CPP, 1STP, 3CPA, and 1PPH) from the Protein Data Bank. In parallel, Monte Carlo with the minimization (MCM) method is also incorporated into the Tinker package for comparison. The energy function, consisting of electrostatic interactions, van der Waals interactions, and torsional energy terms, is calculated using the AMBER94 all-atom empirical force field. Rigid docking simulations for all six complexes and flexible docking simulations for three complexes (1STP, 3CPA, and 1PPH) are carried out using the CSA and the MCM methods. The simulation results show that the docking procedures using the CSA method generally find the most stable complexes as well as the native-like complexes more efficiently and accurately than those using the MCM, demonstrating that CSA is a promising search method for molecular docking problems. PMID- 15538771 TI - Protein microarray system for detecting protein-protein interactions using an anti-His-tag antibody and fluorescence scanning: effects of the heme redox state on protein-protein interactions of heme-regulated phosphodiesterase from Escherichia coli. AB - A highly sensitive microarray system for detecting protein-protein interactions has been developed. This method was successfully applied to analyze the interactions of heme-regulated phosphodiesterase from Escherichia coli (Ec DOS). To immobilize (His)6-Tag fused Ec DOS, anti-(His)6-Tag monoclonal antibody (anti (His)6-Tag mAb) was initially immobilized on the solid surface, and (His)6-Tag fused Ec DOS was fixed by antigen-antibody interactions. For this experiment, ProteoChip, generally suitable for antibody immobilization, was used as solid substrate. In this report, we confirm the antibody immobilization ability of ProteoChip and specific binding to the F(c) region of the antibody. Based on this finding, interdomain interactions between Ec DOS and the isolated heme-bound PAS domain were investigated on the solid surface. Ec DOS immobilized via anti-(His)6 Tag mAb maintained interactions with the PAS fragment, in contrast to directly immobilized Ec DOS in the absence of anti-(His)6-Tag mAb. Heme-redox-sensitive interactions between Ec DOS and the PAS fragment were additionally detected using anti-(His)6-Tag mAb as a mediator. Our results collectively suggest that the immobilization method using anti-Tag antibody is suitable for maintaining native protein characteristics to facilitate elucidation of their structures and functions on solid surfaces. PMID- 15538772 TI - Characterization of a pressurized C5-C16 hydrocarbon gas calibration standard for air analysis. AB - A compressed gas standard containing parts-per-billion (ppb) amounts of the volatile hydrocarbons methylpentadiene (isoprene, 540 ppb) and isooctane (259 ppb) and a series of less volatile C12-C16 n-alkanes (n-dodecane, 349 ppb; n tridecane, 340 ppb; n-tetradecane, 202 ppb; n-pentadecane, 271 ppb; and n hexadecane, 308 ppb) was prepared by a one-step (no further gas dilution) microgravimetric method. The gravimetric mixing ratios were confirmed by referencing to a capillary diffusion method. The cylinder was heated to 75 degrees C to minimize condensation losses of analytes to the cylinder walls. Mixing ratios were monitored over a 2.5-year period. Some initial analyte losses (approximately 3 to 20%) were observed for the heavier C14-C16 n-alkanes. Subsequently, analyte loss rates were found to be in the range of <1.0% per year for n-dodecane to n-hexadecane. The developed guidelines for preparation, storage, and retrieval of these semivolatile analytes enable the use of compressed gas standards for calibration and method development purposes in the environmental gas-phase analysis of these and related compounds. PMID- 15538773 TI - Identification of organic acids in secondary organic aerosol and the corresponding gas phase from chamber experiments. AB - Organic acids in the gas and aerosol phase from photooxidation of 1,3,5 trimethylbenzene in the presence of 300 ppb propene and 300 ppb NOx in smog chamber experiments were determined using a wet effluent diffusion denuder/aerosol collector coupled to ion chromatography (IC) with conductivity detection. Behind the IC, the samples were collected using a fraction collector, for identification of unresolved/unidentified organic acids with IC-mass spectrometry (MS). In total, 20 organic acids were found with MS of which 10 were identified. The organic acids identified offline by IC-MS were then further quantified based on the online IC data. The identification was additionally confirmed with gas chromatography-mass spectrometry. At the maximum aerosol concentration, organic acids comprised 20-45% of the total aerosol mass. The method has a detection limit of 10-100 ng/m3 for the identified carboxylic acids. PMID- 15538774 TI - Sequencing and mass profiling highly modified conotoxins using global reduction/alkylation followed by mass spectrometry. AB - A novel high-throughput method for characterizing heavily modified peptides from cone snail venom is described. Unpurified cone snail duct venom, consisting primarily of multiply disulfide-bonded peptides, is reduced and alkylated using a global procedure in order to simultaneously reduce and derivatize dozens of disulfide-bonded peptides. Samples of Conus victoriae venom are analyzed by online liquid chromatography-electrospray ionization-ion trap-mass spectrometry (LC-ESI-MS) with collisionally induced dissociation (CID). Comparison of the mass profiles of peptides and CID spectra before and after the global reduction and alkylation enables cysteine-containing conopeptides to be ascertained. In this case, over 40 conotoxins are characterized based on only two LC-ESI-MS experiments in terms of mass, number of disulfide-linked cysteine residues (and hence, potential toxin superfamilies), relative hydrophobicity, and other posttranslational modifications. Using this technique, over half of the amino acids (by mass) of several peptides are defined prior to any detailed sequencing studies. Further comparison of the mass data with previously published genetic information allows sequence verification of three novel peptides, termed vc5b, vc6b and vc6c, based on both LC-ESI-MS CID and nanoelectrospray ionization-ion trap-mass spectrometry (nanoESI-MS) experiments. This global method is ideally suited to the use of larger genetic databases in order to efficiently sequence peptides in Conus venoms and is also applicable to analysis of other disulfide rich classes of peptides such as defensins, chemokines, and snake, spider, or other venoms. PMID- 15538775 TI - Determination of polar drug residues in sewage and surface water applying liquid chromatography-tandem mass spectrometry. AB - A simple and rapid method is presented for the trace-level analysis of 10 polar pharmaceutical residues in various types of water samples from the aquatic environment. Using this method, the pharmaceuticals and several drug metabolites can be analyzed in drinking and surface waters and in wastewater (treated and untreated sewage) at concentrations down to 0.01 microg/L. Samples are prepared by a simple in situ derivatization enabling the preconcentration of very polar metabolites by automated solid-phase extraction. The analytes were separated by liquid chromatography with tandem mass spectrometric detection and quantified by comparison with an internal standard. Limits of quantification were between 0.01 and 0.02 microg/L for three phenazone-type pharmaceuticals, six of their metabolites, and the antiepileptic drug carbamazepine. Except for dimethylaminophenazone, recoveries for all analytes were between 87 and 117% for raw and purified sewage, groundwater, and surface and drinking water. Investigations of some environmental samples revealed that sewage and surface water treatment causes a slight reduction of the concentrations of some analytes whereas other compounds were persistent during water treatment. Thus, some compounds were detected at the low-microgram per liter level in sewage effluents of wastewater treatment plants in Berlin (Germany) and were also found at high nanogram per liter concentrations in Berlin surface water samples. PMID- 15538776 TI - Surface plasmon resonance-based sensors to identify cis-regulatory elements. AB - In eukaryotes, transcription is regulated by multiprotein complexes binding to specific regions of genomic DNA, called cis-regulatory elements. Comprehensive identification of these elements is an important goal of functional genomics. Hence, it is of practical interest to develop a high-throughput assay to identify cis-regulatory elements. Toward that goal, we demonstrate that a surface plasmon resonance-based assay can identify whether a specific region of DNA binds to proteins present in raw nuclear lysate. Specifically, we immobilized a 16 basepair double-stranded DNA region of the SQSTM1 promoter to the Texas Instruments Spreeta, a surface plasmon resonance sensor. As a control, in a separate experiment, we immobilized a similar piece of DNA that differed by only a single base pair. We observed a significant difference in surface plasmon resonance signal when these two probes were exposed to raw nuclear lysate from NIH/3T3 cells. Using a luciferase-reporter vector transfected into live NIH/3T3 cells, we measured a significant difference in transcriptional activity between the two pieces of DNA. We conclude that a surface plasmon resonance-based sensor is capable of identifying physiologically significant cis-regulatory elements. PMID- 15538777 TI - Hydrophilic affinity isolation and MALDI multiple-stage tandem mass spectrometry of glycopeptides for glycoproteomics. AB - In glycoproteomics, key structural issues, protein identification, locations of glycosylation sites, and evaluation of the glycosylation site microheterogeneity should be easily evaluated in a large number of glycoproteins, while mass spectrometry (MS) provides substantial information about individual purified glycoproteins. Considering that structural issues are elucidated by studying glycopeptides and that the tandem MS of a tryptic peptide composed of several amino acid residues is enough for protein identification, construction of an MS based method handling tryptic glycopeptides would be of considerable benefit in research. To this end, a simple and efficient method, utilizing hydrophilic binding of carbohydrate matrixes such as cellulose and Sepharose to oligosaccharides, was successfully applied to the isolation of tryptic glycopeptides. Both peptide and oligosaccharide structures were elucidated by multiple-stage tandem MS (MS(n)) of the ions generated by matrix-assisted laser desorption/ionization (MALDI), as follows. The MALDI ion trap mass spectrum of a tryptic glycopeptide mixture from N-linked glycoproteins was composed of the [M + H]+ ions of component glycopeptides. Collision-induced dissociation (CID) of the glycopeptide [M + H]+ ion generated saccharide-spaced peaks, with an interval of, for example, 146, 162, and 203 Da, and their fragment ions corresponding to the peptide and peptide + N-acetylglucosamine (GlcNAc) species in the MS2 spectrum. The saccharide-spaced ladder served to outline oligosaccharide structures, which were then selected as precursors for subsequent MS(n) analyses. The peptide or peptide + GlcNAc ions in the MS2 spectrum or the corresponding ions abundant in the MS1 spectrum were subjected to CID for determination of peptide sequences, to identify proteins and their glycosylation sites. The strategy, isolation of glycopeptides followed by MS(n) analysis, efficiently characterized the structures of beta2-glycoprotein I with four N-glycosylation sites and was applied to an analysis of total serum glycoproteins. PMID- 15538778 TI - Acylic sugar derivatives for GC/MS analysis of 13C-enrichment during carbohydrate metabolism. AB - Metabolic profiling with stable-isotope tracers in combination with gas chromatography/mass spectrometry (GC/MS) is a well-established technique for measuring substrate redistribution within metabolic pathways. This analysis relies on the ability to localize and quantify the fractional incorporation of 13C isotope into each carbon atom of precursor-derived metabolites. In this paper, several carbohydrate derivatization procedures (peracetylation, deuterioalditol acetates, and aldononitrile acetates) are evaluated for the positional isotopic information obtained by gas chromatography/electron impact mass spectrometry (GC/EI-MS). These derivatives have been compared for the quantitative evaluation of 13C distribution into isotopomers of 13C-labeled aldoses and ketoses, and the fragmentation pathways for 15 hexoses, pentoses, and amino sugars of biological origin have been assessed. In addition, a new type of carbohydrate derivative (dialkyldithioacetal acetates) has been developed for GC/MS that retains the charge on the anomeric carbon of the original monosaccharide. Electron impact ionization of these derivatives generates well resolved base peaks arising from C1-C2 bond cleavage with charge retention at the C1 thiol groups. The dialkyldithioacetal acetates are uniquely well suited for measuring isotopic enrichment into the characteristic anomeric carbon of aldose sugars and will facilitate the global analysis of metabolic flux in carbohydrate pathways. PMID- 15538779 TI - Voltammetric determination of the geometrical parameters of inlaid microdisks with shields of thickness comparable to the electrode radius. AB - The cyclic voltammetric behavior of disk microelectrodes surrounded by thin insulating shields (TSM) was investigated from both theoretical and experimental points of view. In particular, microdisks with shields of thickness (b - a), a few electrode radii (a) were considered. A finite difference simulation procedure with a nonuniform, expanding spatial grid, already available in the literature, was employed for predicting shape and height of the voltammograms. The parameters of this numerical simulation were optimized again, and the steady-state limiting currents found in this work for a range of TSMs compared well with previous publications. The steady-state limiting current at TSMs was enhanced with respect to microelectrodes surrounded by thick insulating sheaths (i.e., b >> a), and steady-state conditions were achieved faster. Under these conditions, the difference in potential observed on the forward and backward waves, when the current is half of its maximum value (deltaE(1/2)), was almost equal to zero. Nonsteady-state cyclic voltammograms were also simulated using the optimized parameters, and the effect of scan rate on deltaE(1/2) was examined in detail. Based on this dependence, a voltammetric procedure for the simultaneous determination of microdisk radius and its insulator thickness was proposed. Experimental measurements were performed by using platinum wires, 10-12.5-microm radius, and carbon fibers, 4-microm radius, coated with 4-6-microm-thick electrophoretic paint. The shield thickness produced around the wires was such that b/a < 3. The experimental results obtained were in general congruent with the theory and demonstrated the validity of the method proposed here for the simultaneous determination of radius and shield thickness of a TSM. PMID- 15538780 TI - Monitoring D-serine dynamics in the rat brain using online microdialysis capillary electrophoresis. AB - D-Serine was detected in dialysate collected from the rat striatum using an online microdialysis-CE-LIF instrument. Dialysate can be analyzed every 12.5 s using the online instrument, giving much better temporal resolution than previously possible for D-serine. Basal concentrations of D-serine (8 +/- 2 microM), glutamate (0.8 +/- 0.2 microM), GABA (0.11 +/- 0.04 microM), and L serine (23 +/- 4 microM) were measured. Increases in the concentrations of these neurochemicals induced by the introduction of high-K+ aCSF were quantitated. Notably, an increase in D-serine concentration in response to high-K+ aCSF was observed for the first time. The identity of the D-serine peak was confirmed unambiguously using D-amino acid oxidase to selectively remove D-serine from a dialysate sample. The microdialysis-CE-LIF instrument was able to monitor this enzymatic reaction as it proceeded over a period of 60 min, demonstrating that online microdialysis-CE-LIF is not only useful in monitoring in vivo dynamics but can also be used to monitor other chemical systems. PMID- 15538781 TI - Polymerase chain reaction in high surface-to-volume ratio SiO2 microstructures. AB - We have performed the Taqman beta-actin PCR system in high-surface-to-volume ratio (0.02-0.13 microm(-1)) SiO2 microchannels and observed the reaction yield and uniformity. The concentrations of the enzyme, magnesium chloride, and reaction template were varied in the reaction mix, and PCR amplification was performed in devices of various surface-to-volume ratios. We found that microchannels with higher surface-to-volume ratios required higher enzyme concentrations to achieve the same amplification efficiency. We investigated the possibility that the observed reaction nonuniformity was related to the specific adsorption of magnesium ions to the negatively charged SiO2 surface. The effect of several modifications to the reaction chemistry, the addition of the caged magnesium dye DM-Nitrophen, the replacement of human DNA template with PCR product, and the coating of the microchannel surface with Teflon were all studied. These modifications resulted in improved reaction uniformity in the microchannels and present opportunities for further studies on enhancing the efficiency and uniformity of PCR in high surface-to-volume ratio SiO2 microchannels. PMID- 15538782 TI - Phase-shift fiber-loop ring-down spectroscopy. AB - Fiber-loop ring-down spectroscopy (FLRDS) is a recently developed absorption spectroscopic technique suitable for very small liquid samples. It is based on measurements of the optical decay constant of laser intensity in a loop made of optical waveguide material. This decay constant changes as small liquid samples containing absorbing species are introduced into the loop. In this report, it is demonstrated that one can also obtain the optical decay constant using a continuous wave laser beam that is intensity modulated and then coupled into an optical fiber loop. The inherent exponential decay in the fiber loop introduces a phase shift of the light emitted from the loop with respect to the pumping beam. By measuring this phase shift, one can readily determine the concentration of the analyte introduced between the two fiber ends and a model is established to describe the relationship. It is demonstrated that this technique, dubbed phase shift fiber-loop ring-down spectroscopy (PS-FLRDS), is well suited as an absorption detector for any flow system in which the optical absorption path is limited by the instrument architecture. By measuring the phase angle as a function of concentration of 1,1'-diethyl-4,4'-dicarbocyanine iodide in dimethyl sulfoxide, the detection limit was determined as approximately 6 microM for a 30 40-microm absorption path. A temporal resolution of approximately 100 ms was demonstrated by a rapid displacement of the solutions between the two fiber ends. Proof-of-principle use of the PS-FLRDS detection in capillary flow systems using a commercial four-way microcross established that the alignment of the fiber and the capillary can be made simple and effective, while retaining both a low detection limit and a fast response. PMID- 15538783 TI - Bioassay-directed identification of estrogen residues in urine by liquid chromatography electrospray quadrupole time-of-flight mass spectrometry. AB - A new approach to the search for residues of known and unknown estrogens in calf urine is presented. Following enzymatic deconjugation and solid-phase extraction, a minor part of the samples is screened for estrogen activity using a recently developed rapid reporter gene bioassay. The remainder of the bioactive extracts is analyzed by gradient liquid chromatography (LC) with, in parallel, bioactivity and mass spectrometric detection via effluent splitting toward a 96-well fraction collector and an electrospray quadrupole time-of-flight mass spectrometer (QTOFMS). The LC fractions in the 96-well plate are used for the detection of estrogen activity using the bioassay. The biogram obtained features a 20-s time resolution, and the suspect well numbers can be easily correlated with the LC/QTOFMS retention time. The mass spectral data from the thus assigned relevant parts of the chromatograms are background subtracted, followed by accurate mass measurement, element composition calculation, and identification. The method allows estrogen activity detection and identification of (un)known estrogens in urine at the 1-2 ng/L level, in compliance with current residue analysis performance for hormone abuse in cattle. The applicability of this LC/bioassay/QTOFMS approach for the identification of estrogens in real-life samples is demonstrated by the analysis of several calf urine samples, and preliminary data from a pig feed sample. PMID- 15538784 TI - Bacterial identification by protein mass mapping combined with an experimentally derived protein mass database. AB - A protein mass mapping approach using mass spectrometry (MS) combined with an experimentally derived protein mass database is presented for rapid and effective identification of bacterial species. A prototype mass database from the protein extracts of nine bacterial species has been created by off-line high-performance liquid chromatography (HPLC) matrix-assisted laser desorption/ionization (MALDI) MS, in which the microbiological parameter of bacterial growth time is considered. A numerical method using a statistical weight factor algorithm is devised for matching the protein masses of an unknown bacterial sample against the database. The sum of these weight factors produces a corresponding summed weight factor score for each bacterial species listed in the database, and the database species producing the highest score represents the identity of the respective unknown bacterium. The applicability and reliability of this protein mass mapping approach has been tested with seven bacterial species in a single blind study by both direct MALDI MS and HPLC electrospray ionization MS methods, and identification results with 100% accuracy are obtained. Our studies have demonstrated that the protein mass database can be rapidly established and readily adopted with relatively less dependency on experimental factors. Furthermore, it is shown that a number of proteins can be detected using a protein sample amount equivalent to an extract of less than 1000 cells, demonstrating that this protein mass mapping approach can potentially be highly sensitive for rapid bacterial identification. PMID- 15538785 TI - N-terminal isotope tagging strategy for quantitative proteomics: results-driven analysis of protein abundance changes. AB - Comparing the relative abundance of each protein present in two or more complex samples can be accomplished using isotope-coded tags incorporated at the peptide level. Here we describe a chemical labeling strategy for the incorporation of a single isotope label per peptide, which is completely sequence-independent so that it potentially labels every peptide from a protein including those containing posttranslational modifications. It is based on a gentle chemical labeling strategy that specifically labels the N-terminus of all peptides in a digested sample with either a d5- or d0-propionyl group. Lysine side chains are blocked by guanidination prior to N-terminal labeling to prevent the incorporation of multiple labels. In this paper, we describe the optimization of this N-terminal isotopic tagging strategy and validate its use for peptide-based protein abundance measurements with a 10-protein standard mixture. Using a results-driven strategy, which targets proteins for identification based on MALDI TOF-MS analysis of isotopically labeled peptide pairs, we also show that this labeling strategy can detect a small number of differentially expressed proteins in a mixture as complex as a yeast cell lysate. Only peptides that show a difference in relative abundance are targeted for identification by tandem MS. Despite the fact that many peptides are quantitated, only those few showing a difference in abundance are targeted for protein identification. Proteins are identified by either targeted LC-ES MS/MS or MALDI TOF/TOF. Identifications can be accomplished equally well by either technique on the basis of multiple peptides. This increases the confidence level for both identification and quantitation. The merits of ES MS/MS or MALDI MS/MS for protein identification in a results-driven strategy are discussed. PMID- 15538786 TI - Simple identification of a cross-linked hemoglobin by tandem mass spectrometry in human serum. AB - Hemoglobin-based oxygen therapeutics are prepared by reaction of hemoglobin with cross-linking molecules and are utilized as blood substitutes. They can be used as doping agents to increase the oxygen-carrying capacity of hemoglobin. We have compared a glutaraldehyde-polymerized bovine hemoglobin (Oxyglobin, Biopure Corp.) with natural bovine hemoglobin by mass spectrometry in order to detect specific fragment ions of the cross-linked protein for further potential applications in doping control of human blood samples. HCl acid (6 N) hydrolysis was performed in parallel on both proteins. Hydrolysates were then analyzed by direct infusion electrospray mass spectrometry (ESIMS) using a triple quadrupole mass spectrometer. Confirmation and precision were obtained by LC-ESIMS(n) experiments performed on an ion trap mass spectrometer. Chromatographic and mass spectrometry data allowed detection of two potential Oxyglobin-specific ions--m/z 299 and 399--that were shown to lose a 159 u neutral fragment under collision induced dissociation conditions. Thus, monitoring of constant neutral loss of 159 u on acid hydrolysates of human serum samples spiked with different amounts of Oxyglobin has proved to be an efficient screening method to specifically detect and identify Oxyglobin. LC-MS of the spiked serum sample hydrolysates enabled detection of Oxyglobin at a detection limit of 4 g x L(-1). PMID- 15538787 TI - Determination of selenomethionine and selenocysteine in human serum using speciated isotope dilution-capillary HPLC-inductively coupled plasma collision cell mass spectrometry. AB - A method for the accurate determination of selenoamino acids in human serum by HPLC-ICPMS was developed using the species-specific isotope dilution analysis principle. A serum sample was enzymatically digested with a mixture of lipase and protease after derivatization of the selenocysteine residues with iodoacetamide. The selenoamino acid fraction was isolated by size exclusion LC followed by the separation of selenomethionine and the carboxymethylated selenocysteine by capillary HPLC. The isotope-specific determination of 77Se and 80Se was achieved on-line by ICP collision cell MS allowing the removal of polyatomic interferences. Quantification was carried out by isotope dilution using a 77Se labeled selenomethionine spike and the determination of the 77Se/80Se ratio in the cHPLC selenomethionine peak. The accurately determined selenomethionine was used as an internal standard for the selenocysteine determination from the same chromatogram. The modification of the previously developed cHPLC-ICPMS interface allowed the reduction of the absolute detection limits twice (down to the 75-fg level), which resulted in the lowest ever reported procedural detection limits (below 0.5 ng g(-1) for a 450-mg serum sample). The precision was less than 5% RSD. The method was validated by the mass balance of selenium (amino acid incorporated vs total). PMID- 15538788 TI - Photoimmobilization of proteins for affinity capture combined with MALDI TOF MS analysis. AB - Affinity capture surfaces can be prepared in a number of ways. A method of obtaining such surfaces through UV-activated immobilization of binding proteins using a benzophenone derivative is reported. Photoimmobilized protein G was used to selectively capture and preconcentrate bovine IgG from a mixture with BSA, and the affinity of photoattached concanavalin A toward ovalbumin was compared with that of commercially available concanavalin A on agarose beads. The results of the capture after tryptic digestion were analyzed by MALDI TOF MS. Immobilized trypsin was also prepared through photoimmobilization and later used to digest hemoglobin. Immobilized enzyme digestion resulted in more partial cleavages than solution-phase digestion. More methionine and tryptophan oxidation was also observed. Photoimmobilization was shown to be a quick and easy way of immobilizing ligands on surfaces. PMID- 15538789 TI - Depth profiling of Langmuir-Blodgett films with a buckminsterfullerene probe. AB - Bombardment with C60+ primary ions of monolayer and multilayer barium arachidate Langmuir-Blodgett (LB) films is investigated. The behavior of cluster versus atomic (Ga+) bombardment is monitored by the barium-cationized arachidate ion (mass-to-charge ratio (m/z) 449) and a characteristic fragment ion (m/z 209) using 1-, 7-, and 15-layer model systems. The removal rate of material from the films is shown to be on the order of several hundred molecules per C60 impact, a value 100-fold larger than Ga+ impact. The enhancement in secondary ion yield is also shown to be larger for the 15-layer film (400x) than for the monolayer film (100x). Moreover, most of the increase in yield is shown to be associated with ejection of sputtered species rather than an increase in ionization probability. High yields associated with cluster bombardment are also shown to be amenable to depth profiling experiments in which the two ions can be monitored as the film is being removed. In this modality, chemical damage associated with bombardment is removed before it can accumulate on the surface. Due to the similarity of fatty acid LB films to cellular membranes, these results suggest that C60+ primary ion beams may improve the prospects for TOF-SIMS studies of biological systems. PMID- 15538790 TI - Microchip atmospheric pressure chemical ionization source for mass spectrometry. AB - A novel microchip heated nebulizer for atmospheric pressure chemical ionization mass spectrometry is presented. Anisotropic wet etching is used to fabricate the flow channels, inlet, and nozzle on a silicon wafer. An integrated heater of aluminum is sputtered on a glass wafer. The two wafers are jointed by anodic bonding, creating a two-dimensional version of an APCI source with a sample channel in the middle and gas channels symmetrically on both sides. The ionization is initiated with an external corona-discharge needle positioned 2 mm in front of the microchip heated nebulizer. The microchip APCI source provides flow rates down to 50 nL/min, stable long-term analysis with chip lifetime of weeks, good quantitative repeatability (RSD < 10%) and linearity (r(2) > 0.995) with linear dynamic rage of at least 4 orders of magnitude, and cost-efficient manufacturing. The limit of detection (LOD) for acridine measured with microchip APCI at flow rate of 6.2 muL/min was 5 nM, corresponding to a mass flow of 0.52 fmol/s. The LOD with commercial macro-APCI at a flow rate of 1 mL/min for acridine was the same, 5 nM, corresponding to a significantly worse mass flow sensitivity (83 fmol/s) than measured with microchip APCI. The advantages of microchip APCI makes it a very attractive new microfluidic detector. PMID- 15538791 TI - Programmed field decay thermal field flow fractionation of polymers: a calibration method. AB - The universal calibration procedure typical of thermal field flow fractionation (ThFFF) under constant thermal field operation was extended to thermal field programming (TFP) operation. The method requires knowledge of the following: (a) the programming function, which only depends on the thermal field decay function, (b) the physicochemical properties of the solvent, and (c) the calibration plot under varying channel cold wall temperatures (T(c)). Two field flow fractionation field programming conditions, with either a constant or a variable in time carrier flow velocity, are exploited. The method is based on determination, for each retention time position, of the average lambda retention value typical of TFP ThFFF. This parameter is then used to obtain the calibration plot (i.e., the molecular weight of the species as a function of the retention time position) by using the programming function and the calibration plot under varying T(c) values. The procedure approximation errors are also derived as a function of the programming type and solute-solvent system. To properly test the procedure, the calibration plot for the system constituted by polystyrene (PS) in cis-trans Decalin was determined, under varying conditions T(c) and thermal gradients, by using a set of monodisperse PS standards of different molecular weights (M). The procedure was first validated by simulation under two typical cases of TFP ThFFF operation. The approximation errors were found acceptable (in the worse cases, the accuracy in M prediction was 3%) and are in agreement with the theory. The procedure was then experimentally validated under varying programming decay function conditions. The reproducibility and accuracy of the M determination are both better than 2%. PMID- 15538792 TI - Chiral separations using a polypeptide and polymeric dipeptide surfactant polyelectrolyte multilayer coating in open-tubular capillary electrochromatography. AB - A polyelectrolyte multilayer (PEM) coating consisting of the polypeptide, poly(l lysine) hydrobromide, poly(l-lysine) and the polymeric dipeptide surfactant, poly(sodium undecanoyl-l-leucyl-alaninate), poly(l-SULA), is investigated as a new medium for the separation of chiral analytes in open-tubular capillary electrochromatography (OT-CEC). In this approach, a stable PEM is constructed in situ by alternative rinses of the cationic polymer poly(l-lysine) and the anionic polymer poly(l-SULA). In previous studies, the PEM coating has been constructed by use of the cationic polyelectrolyte poly (diallydimethylammonium chloride), PDADMAC. In this study, we investigate the use of a biopolymer as the cationic polyelectrolyte. The results reported here indicate an increase in selectivity and resolution when poly(l-lysine) is used as the cationic polymer in place of PDADMAC. To evaluate the chromatographic performance of the PEM coating as a chiral stationary phase, the separation of the beta-blockers, labetalol and sotalol, and the binaphthyl derivatives, 1,1'-bi-2-naphthyl-2,2'-dihydrogen phosphate, 1,1'-bi-2-naphthol, and 1,1-binaphthyl-2,2'-diamine, are investigated. In addition, the effect of varying the amino acid order of the polymeric dipeptide surfactant on resolution is investigated. The number of bilayers also significantly influences the separation efficiency and resolution of enantiomers. The run-to-run and capillary-to-capillary reproducibilities are evaluated by calculating the relative standard deviations (RSDs) of the electroosmotic flow. These RSD values were found to be less than 1%. The coating is also stable and allows more than 290 runs to be performed in the same capillary. In addition, coupling of this chiral OT-CEC column with mass spectrometry is investigated. PMID- 15538793 TI - Chip-based bioassay using bacterial sensor strains immobilized in three dimensional microfluidic network. AB - A whole-cell bioassay has been performed using Escherichia coli sensor strains immobilized in a chip assembly, in which a silicon substrate is placed between two poly(dimethylsiloxane) (PDMS) substrates. Microchannels fabricated on the two separate PDMS layers are connected via perforated microwells on the silicon chip, and thus, a three-dimensional microfluidic network is constructed in the assembly. Bioluminescent sensor strains mixed with agarose are injected into the channels on one of the two PDMS layers and are immobilized in the microwells by gelation. Induction of the firefly luciferase gene expression in the sensor strains can be easily carried out by filling the channels on the other layer with sample solutions containing mutagen. Bioluminescence emissions from each well are detected after injection of luciferin/ATP mixtures into the channels. In this assay format using two multichannel layers and one microwell array chip, the interactions between various types of samples and strains can be monitored at each well on one assembly in a combinatorial fashion. Using several genotypes of the sensor strains or concentrations of mitomycin C in this format, the dependence of bioluminescence on these factors was obtained simultaneously in the single screening procedure. The present method could be a promising on-chip format for high-throughput whole-cell bioassays. PMID- 15538794 TI - Microdeposition device interfacing capillary electrochromatography and microcolumn liquid chromatography with matrix-assisted laser desorption/ionization mass spectrometry. AB - A sample deposition device has been constructed and optimized for interfacing CEC and capillary LC columns to MALDI mass spectrometry. For CEC analysis, the device is composed of an inlet buffer reservoir and an outlet buffer reservoir connected to a matrix reservoir through a connection sleeve. The matrix reservoir is connected to a deposition capillary via another connection sleeve. CEC eluent is transported to the matrix reservoir via a capillary that is connected to the deposition capillary by the connection sleeve inside the matrix reservoir. This connection sleeve also acts as a mixing chamber, allowing the CEC eluent to be mixed with matrix prior to deposition. Complex glycan mixtures can be separated by CEC using hydrophilic-phase monolithic columns, with capillary eluent being deposited on a standard MALDI plate along with a suitable matrix solution. Thousands of discrete, highly homogeneous dots can be generated for a subsequent mass spectrometric analysis. With minor modifications, this device is also applicable to capillary LC of peptides using gradient elution. In this configuration, the outlet of the LC column is connected to a deposition capillary inside a matrix reservoir through a connection sleeve that allows mixing of the LC effluent with an appropriate matrix. The device has been evaluated with the tryptic digests of proteins. PMID- 15538795 TI - General rules for the optimal external porosity of LC supports. AB - We present a series of numerically calculated plate height and flow resistance data obtained for an idealized chromatographic support mimic with variable bed porosity (0.3 4 microg/ml; intermediate, 8 to 16 microg/ml; resistant, > 32 microg/ml. 5FC was very active against Candida spp. and other medically important yeasts as 852 (83.4%) of the studied isolates were susceptible (MIC < 4 microg/ml). The species most susceptible to 5FC were C. dubliniensis (100%of isolates; MIC90, 0.25 microg/ml), C. famata (100% of isolates; MIC90, 0.25 microg/ml), C. guilliermondii (98%of isolates; MIC90, 0.25 microg/ml), C. glabrata (95.5% of isolates; MIC90, 0.25 microg/ml), and C. neoformans (90.9% of isolates; MIC90, 2 microg/ml). Primary resistance to 5FC was very uncommon, and a MIC > 32 microg/ml, indicator of in vitro resistance, was observed in 106 isolates (10.4%): 77 C. albicans (16.5% of isolates; MIC90, > 128 microg/ml), 9 C. parapsilosis (6.4% of isolates; MIC90, 8 microg/ml), 4 C. albidus (80% of isolates, MIC50, > 128 microg/ml), 3 C. glabrata (4.4% of isolates; MIC90, 0.25 microg/ml), 3 C. tropicalis (8.8% of isolates; MIC90, 4 microg/ml), 2 C. krusei (7.1% of isolates; MIC90, 8 microg/ml), 2 Rhodotorula spp. (4.6% of isolates, MIC90, 1 microg/ml), 8 Trichosporon spp. (33.3% of isolates; MIC90, 64 microg/ml), and 1 C. lipolytica (50% of isolates). Interestingly, most C. albicans (67 out of 77 isolates) resistant to 5FC were serotype B isolates. PMID- 15538830 TI - Differences in extracellular enzymatic activity between Candida dubliniensis and Candida albicans isolates. AB - Twenty-six Candida dubliniensis and 27 Candida albicans oral strains isolated from patients infected by the human immunodeficiency virus (HIV) were tested for germ tube production and 21 extracellular enzymatic activities. Assessment of the enzymatic profile was performed by using the API-ZYM commercial kit system (bioMerieux, France), which tests 19 different enzymes. Protease activity was expressed during the first days of incubation by 100% of the strains studied and resulted higher than phospholipase activity in the C. dubliniensis and C. albicans strains tested. The API-ZYM profile of the C. dubliniensis and C. albicans strains differs with respect to the number and percentage of the enzymes considered, as well as with the intensity of the substrate metabolized by the strains, in particular for the enzymes n 8 (cystine-arylamidase), n 12 (naphtol AS-BI-phosphohydrolase) and n 16 (alpha-glucosidase). These enzymes may be useful to differentiate C. dubliniensis and C. albicans together with other phenotypic characteristics proposed in the literature. No relationship among protease, phospholipase and other extracellular enzymatic activities was observed in C. dubliniensis. The average percentage of strains filamentation after 4 h was between 32 and 42%. PMID- 15538831 TI - [Interruption of antifungal secondary prophylaxis in AIDS-related histoplasmosis]. AB - The clinical data of 21 patients, suffering AIDS-related histoplasmosis, who were able to interrupt antifungal secondary prophylaxis, after achieving a partial restoration of the cell mediated immunity by HAART administration, are presented. They were 16 males and five females, whose ages varied between 32 and 54 years (mean = 38.5 years). All of them presented disseminated progressive forms of histoplasmosis, with multiple locations (skin, mucous membranes, liver, spleen, lymph nodes and lungs). The majority of the cases suffered other concomitant diseases (specially tuberculosis and Kaposi sarcoma), 66.6 % of the patients had less than 50 CD4+ cells/microl at the start of treatment and the average viral burden was 278,385 RNA copies/ml. The initial treatment consisted in 400 mg/day of itraconazole, by oral route, in 14 cases and the remaining seven patients were treated with amphotericin B, intravenously, at a daily dose of 0.7 mg/kg of body weight. One patient who did not tolerate amphotericin B and presented a partial response to itraconazole, was treated with posaconazole orally at a daily dose of 800 mg. Fourteen patients received oral itraconazole at a daily dose of 200 mg as a secondary prophylaxis, the remaining three patients were treated with intravenous amphotericin B, 50 mg twice a week. After HAART for an average lapse of 16.7 months (10 to 32 months), five cases showed CD4+ cells counts above 150 cells/microl and the remaining 16 presented more than 200 cells/microl; 18 of them had undetectable viral burden and all cases were asymptomatic. The follow up after secondary prophylaxis discontinuation varied between six months and six years (mean= 33.6 months). Twenty out of 21 patients (95 %) were clinically stable, without any manifestation of relapses, including two patients who abandoned HAART. One patient, who discontinued HAART, contracted a fatal bacterial pneumonia. Even though the limited number of cases, the data presented in this study seem to suggest that it is possible to interrupt antifungal secondary prophylaxis of histoplasmosis, when the patient is clinically asymptomatic and the CD4+ cells counts are above 150 cells/microl. PMID- 15538832 TI - Posaconazole therapy for severe abdominal candidiasis: a case report. AB - We report the successful treatment of a fluconazole-resistant intra-abdominal Candida infection (Candida albicans and Candida tropicalis) with posaconazole (SCH56592) in a 68-year-old woman with a recent history of intra-abdominal surgery. PMID- 15538833 TI - [Rapid identification of Candida glabrata using a new commercial kit]. AB - Candida glabrata is an emergent pathogen with diminished susceptibility to azoles, thus a rapid identification of this yeast could be of help to choose the appropriate treatment. GLABRATA RTT (Fumouze Diagnostics, France) is a new C. glabrata identification test. To evaluate its utility in the clinical laboratory daily routine, we prospectively tested 168 yeasts isolated in our hospital. GLABRATA RTT results had a sensitivity of 98.4% and a specificity of 100%. The combination of CHROMagar Candida isolation medium and GLABRATA RTT test allowed the identification of the four most common species in the clinical practice (Candida albicans, Candida tropicalis, Candida krusei and C. glabrata). PMID- 15538834 TI - [Candida ciferrii in an immunocompromised patient]. AB - A case of possible infection due to Candida ciferrii in an immunocompromised patient is presented. This fungal species has been rarely reported as cause of human infection. The isolate showed in vitro resistance to fluconazole. PMID- 15538835 TI - A new caffeic acid minimal synthetic medium for the rapid identification of Cryptococcus neoformans isolates. AB - Melanin production is one of the most important criteria for rapid identification of Cryptococcus neoformans. Most of the media described in the literature for identifying C. neoformans are very complex; they contain many organic or inorganic compounds and are difficult to prepare and store. The new minimal synthetic caffeic acid medium described in this paper is simpler to prepare, convenient and constitutes an interesting new medium for the rapid identification of C. neoformans isolates. PMID- 15538836 TI - [Carbohydrate assimilation by clinical and environmental Rhodotorula glutinis strains]. AB - This study was carried to determine the carbohydrate assimilation patterns of Rhodotorula strains isolated from clinical and environmental specimens. We have tested the commercial system ID 32C (bioMerieux, France) on 80 different strains of Rhodotorula glutinis: 47 strains from clinical samples and 33 strains from environmental samples. The assimilation percentages obtained in our study for galactose, cellobiose, gluconate and sorbose were lower than those showed in the identification table of the method. However, the assimilation percentages for mannitol and esculin were higher. According to our results, we conclude that the numerical profiles and the identification software of the commercial system present limitations for the characterization of some R. glutinis strains. PMID- 15538837 TI - Tinea facei due to Trichophyton soudanense: first report in Argentina. AB - We hereby report the first case of a tinea faciae infection due to Trichophyton soudanense in a 20-year-old female from Buenos Aires city, Argentina. This case illustrates the need to be aware of unusual agents and the importance of a proper mycological diagnosis. PMID- 15538838 TI - [Growth in species of the genus Ascobolus. II. (Pezizales-Ascomycota)]. AB - The kinetics of growth of six heterothallic species of the genus Ascobolus was studied in liquid culture media. The results obtained showed variation among the species in the duration of the different phases of the growth cycle. Four groups can be recognized considering the extension of the exponential phase of growth. The stationary phase, which differs in its length, is frequently very short, entering quickly in the phase of death, accompanied by the autolysis of the mycelium. PMID- 15538839 TI - [Clinical problems in medical mycology: case No. 9. Paracoccidioidomycosis]. PMID- 15538840 TI - The ground state tunneling splitting of malonaldehyde: accurate full dimensional quantum dynamics calculations. AB - Benchmark calculations of the tunneling splitting in malonaldehyde using the full dimensional potential proposed by Yagi et al. are reported. Two exact quantum dynamics methods are used: the multiconfigurational time-dependent Hartree (MCTDH) approach and the diffusion Monte Carlo based projection operator imaginary time spectral evolution (POITSE) method. A ground state tunneling splitting of 25.7+/-0.3 cm(-1) is calculated using POITSE. The MCTDH computation yields 25 cm(-1) converged to about 10% accuracy. These rigorous results are used to evaluate the accuracy of approximate dynamical approaches, e.g., the instanton theory. PMID- 15538841 TI - Full configuration interaction potential energy curves for the X (1)Sigma(g) (+), B (1)Delta(g), and B(') (1)Sigma(g) (+) states of C(2): a challenge for approximate methods. AB - The C(2) molecule exhibits unusual bonding and several low-lying excited electronic states, making the prediction of its potential energy curves a challenging test for quantum chemical methods. We report full configuration interaction results for the X (1)Sigma(g) (+), B (1)Delta(g), and B(') (1)Sigma(g) (+) states of C(2), which exactly solve the electronic Schrodinger equation within the space spanned by a 6-31G( *) basis set. Within the D(2h) subgroup used by most electronic structure programs, these states all have the same symmetry ((1)A(g)), and all three states become energetically close for interatomic distances beyond 1.5 A. The quality of several single-reference ab initio methods is assessed by comparison to the benchmark results. Unfortunately, even coupled-cluster theory through perturbative triples using an unrestricted Hartree-Fock reference exhibits large nonparallelity errors (>20 kcal mol(-1)) for the ground state. The excited states are not accurately modeled by any commonly used single-reference method, nor by configuration interaction including full quadruple substitutions. The present benchmarks will be helpful in assessing theoretical methods designed to break bonds in ground and excited electronic states. PMID- 15538842 TI - An efficient method for calculating maxima of homogeneous functions of orthogonal matrices: applications to localized occupied orbitals. AB - We present here three new algorithms (one purely iterative and two DIIS-like [Direct Inversion in the Iteractive Subspace]) to compute maxima of homogeneous functions of orthogonal matrices. These algorithms revolve around the mathematical lemma that, given an invertible matrix A, the function f(U)=Tr(AU) has exactly one local (and global) maximum for U special orthogonal [i.e., UU(T)=1 and det(U)=1]. This is proved in the Appendix. One application of these algorithms is the computation of localized orbitals, including, for example, Boys and Edmiston-Ruedenberg (ER) orbitals. The Boys orbitals are defined as the set of orthonormal orbitals which, for a given vector space of orbitals, maximize the sum of the distances between orbital centers. The ER orbitals maximize total self interaction energy. The algorithm presented here computes Boys orbitals roughly as fast as the traditional method (Jacobi sweeps), while, for large systems, it finds ER orbitals potentially much more quickly than traditional Jacobi sweeps. In fact, the required time for convergence of our algorithm scales quadratically in the region of a few hundred basis functions (though cubicly asymptotically), while Jacobi sweeps for the ER orbitals traditionally scale as the number of occupied orbitals to the fifth power. As an example of the utility of the method, we provide below the ER orbitals of nitrated and nitrosated benzene, and we discuss the chemical implications. PMID- 15538843 TI - Coping with the node problem in quantum hydrodynamics: the covering function method. AB - A conceptually simple approach, the covering function method (CFM), is developed to cope with the node problem in the hydrodynamic formulation of quantum mechanics. As nodes begin to form in a scattering wave packet (detected by a monitor function), a nodeless covering wave function is added to it yielding a total function that is also nodeless. Both local and global choices for the covering function are described. The total and covering functions are then propagated separately in the hydrodynamic picture. At a later time, the actual wave function is recovered from the two propagated functions. The results obtained for Eckart barrier scattering in one dimension are in excellent agreement with exact results, even for very long propagation times t=1.2 ps. The capability of the CFM is also demonstrated for multidimensional propagation of a vibrationally excited wave packet. PMID- 15538844 TI - Density functional response theory calculations of three-photon absorption. AB - Three-photon absorption probabilities delta(3PA) have been calculated through application of a recently derived method for cubic response functions within density functional theory (DFT). Calculations are compared with Hartree-Fock (HF) and with a coupled cluster hierarchy of models in a benchmarking procedure. Except for cases having intermediate states near resonance, density functional theory is demonstrated to be in sufficient agreement with the highly correlated methods in order to qualify for predictions of delta(3PA). For the larger systems addressed, a set of acceptor A and donor D substituted pi-conjugated systems formed by trans-stilbene and dithienothiophene (DTT), we find noticeable differences in the magnitude of delta(3PA) between HF and DFT, although similar trends are followed. It is shown that the dipolar structures, TS-AD and DTT-AD, have substantially larger delta(3PA) than other types of modifications which is in accordance with observations for two-photon absorption. This is the first application of density functional theory to three-photon absorption beyond the use of few-state models. PMID- 15538845 TI - Quantum statistical mechanics with Gaussians: equilibrium properties of van der Waals clusters. AB - The variational Gaussian wave-packet method for computation of equilibrium density matrices of quantum many-body systems is further developed. The density matrix is expressed in terms of Gaussian resolution, in which each Gaussian is propagated independently in imaginary time beta=(k(B)T)(-1) starting at the classical limit beta=0. For an N-particle system a Gaussian exp[(r-q)(T)G(r q)+gamma] is represented by its center qinR(3N), the width matrix GinR(3Nx3N), and the scale gammainR, all treated as dynamical variables. Evaluation of observables is done by Monte Carlo sampling of the initial Gaussian positions. As demonstrated previously at not-very-low temperatures the method is surprisingly accurate for a range of model systems including the case of double-well potential. Ideally, a single Gaussian propagation requires numerical effort comparable to the propagation of a single classical trajectory for a system with 9(N(2)+N)/2 degrees of freedom. Furthermore, an approximation based on a direct product of single-particle Gaussians, rather than a fully coupled Gaussian, reduces the number of dynamical variables to 9N. The success of the methodology depends on whether various Gaussian integrals needed for calculation of, e.g., the potential matrix elements or pair correlation functions could be evaluated efficiently. We present techniques to accomplish these goals and apply the method to compute the heat capacity and radial pair correlation function of Ne(13) Lennard-Jones cluster. Our results agree very well with the available path integral Monte Carlo calculations. PMID- 15538846 TI - Calculation of excited-state properties using general coupled-cluster and configuration-interaction models. AB - Using string-based algorithms excitation energies and analytic first derivatives for excited states have been implemented for general coupled-cluster (CC) models within CC linear-response (LR) theory which is equivalent to the equation-of motion (EOM) CC approach for these quantities. Transition moments between the ground and excited states are also considered in the framework of linear-response theory. The presented procedures are applicable to both single-reference-type and multireference-type CC wave functions independently of the excitation manifold constituting the cluster operator and the space in which the effective Hamiltonian is diagonalized. The performance of different LR-CC/EOM-CC and configuration-interaction approaches for excited states is compared. The effect of higher excitations on excited-state properties is demonstrated in benchmark calculations for NH(2) and NH(3). As a first application, the stationary points of the S(1) surface of acetylene are characterized by high-accuracy calculations. PMID- 15538847 TI - Cooperating or fighting with control noise in the optimal manipulation of quantum dynamics. AB - This paper investigates the impact of control field noise on the optimal manipulation of quantum dynamics. Simulations are performed on several multilevel quantum systems with the goal of population transfer in the presence of significant control noise. The noise enters as run-to-run variations in the control amplitude and phase with the observation being an ensemble average over many runs as is commonly done in the laboratory. A genetic algorithm with an improved elitism operator is used to find the optimal field that either fights against or cooperates with control field noise. When seeking a high control yield it is possible to find fields that successfully fight with the noise while attaining good quality stable results. When seeking modest control yields, fields can be found which are optimally shaped to cooperate with the noise and thereby drive the dynamics more efficiently. In general, noise reduces the coherence of the dynamics, but the results indicate that population transfer objectives can be met by appropriately either fighting or cooperating with noise, even when it is intense. PMID- 15538848 TI - An extension of the quasicontinuum treatment of multiscale solid systems to nonzero temperature. AB - Covering the solid lattice with a finite-element mesh produces a coarse-grained system of mesh nodes as pseudoatoms interacting through an effective potential energy that depends implicitly on the thermodynamic state. Use of the pseudoatomic Hamiltonian in a Monte Carlo simulation of the two-dimensional Lennard-Jones crystal yields equilibrium thermomechanical properties (e.g., isotropic stress) in excellent agreement with "exact" fully atomistic results. PMID- 15538849 TI - Multidimensional density operator propagations in open systems: model studies on vibrational relaxations and surface sticking processes. AB - An efficient method for the numerical treatment of multidimensional dynamics of open systems is presented: the multiconfiguration time-dependent Hartree (MCTDH) method extended to the propagation of density operators. With this method we investigate the relaxation process of a CO molecule adsorbed on a copper surface, i.e., CO/Cu(100), after the excitation with an infrared (IR) pulse. The interaction potential was taken from the literature. Lifetime estimations and thermalization studies were performed on the IR excited CO molecule. We were able to treat this system with all six degrees of freedom (DOF) and thus 12 dynamical variables, but most of our studies used a two or four DOF model. Finally, we demonstrate the applicability of MCTDH to the analysis of scattering processes in an open environment. We calculate sticking coefficients of a scattered particle to a model surface, the latter acting as heat bath. The surface corrugation and the initial particle energy have been varied, and six different relaxation strengths have been studied. These calculations were done under the inclusion of three DOFs: the two surface coordinates and the distance between the particle and the surface. PMID- 15538850 TI - Frame transformation relations for fluxional symmetric rotor dimers. AB - The theory of frame transformation relation connecting body oriented angular momentum states and lab weakly coupled momentum states have been extended from rotor-electron to rotor-dimer systems. Coupling schemes are analyzed for weak and strong cases of correlation between lab and two different rotor body frames. It is shown that the frame transformation relation is a purely quantum effect at low angular momentum but an approach to a classical limit for high J. Symmetry analysis of frame transformation is compared to eigensolutions of model coupling Hamiltonian. PMID- 15538851 TI - Efficient calculation of potential energy surfaces for the generation of vibrational wave functions. AB - An automatic procedure for the generation of potential energy surfaces based on high level ab initio calculations is described. It allows us to determine the vibrational wave functions for molecules of up to ten atoms. Speedups in computer time of about four orders of magnitude in comparison to standard implementations were achieved. Effects due to introduced approximations--within the computation of the potential--on fundamental modes obtained from vibrational self-consistent field and vibrational configuration interaction calculations are discussed. Benchmark calculations are provided for formaldehyde and 1,2,5-oxadiazole (furazan). PMID- 15538852 TI - Atomic spectral methods for molecular electronic structure calculations. AB - Theoretical methods are reported for ab initio calculations of the adiabatic (Born-Oppenheimer) electronic wave functions and potential energy surfaces of molecules and other atomic aggregates. An outer product of complete sets of atomic eigenstates familiar from perturbation-theoretical treatments of long range interactions is employed as a representational basis without prior enforcement of aggregate wave function antisymmetry. The nature and attributes of this atomic spectral-product basis are indicated, completeness proofs for representation of antisymmetric states provided, convergence of Schrodinger eigenstates in the basis established, and strategies for computational implemention of the theory described. A diabaticlike Hamiltonian matrix representative is obtained, which is additive in atomic-energy and pairwise atomic interaction-energy matrices, providing a basis for molecular calculations in terms of the (Coulombic) interactions of the atomic constituents. The spectral product basis is shown to contain the totally antisymmetric irreducible representation of the symmetric group of aggregate electron coordinate permutations once and only once, but to also span other (non-Pauli) symmetric group representations known to contain unphysical discrete states and associated continua in which the physically significant Schrodinger eigenstates are generally embedded. These unphysical representations are avoided by isolating the physical block of the Hamiltonian matrix with a unitary transformation obtained from the metric matrix of the explicitly antisymmetrized spectral-product basis. A formal proof of convergence is given in the limit of spectral closure to wave functions and energy surfaces obtained employing conventional prior antisymmetrization, but determined without repeated calculations of Hamiltonian matrix elements as integrals over explicitly antisymmetric aggregate basis states. Computational implementations of the theory employ efficient recursive methods which avoid explicit construction the metric matrix and do not require storage of the full Hamiltonian matrix to isolate the antisymmetric subspace of the spectral-product representation. Calculations of the lowest-lying singlet and triplet electronic states of the covalent electron pair bond (H(2)) illustrate the various theorems devised and demonstrate the degree of convergence achieved to values obtained employing conventional prior antisymmetrization. Concluding remarks place the atomic spectral-product development in the context of currently employed approaches for ab initio construction of adiabatic electronic eigenfunctions and potential energy surfaces, provide comparisons with earlier related approaches, and indicate prospects for more general applications of the method. PMID- 15538853 TI - Dynamics of (H(-), H(2)) collisions: a time-dependent quantum mechanical investigation on a new ab initio potential energy surface. AB - A global analytical potential energy surface for the ground state of H(3)(-) has been constructed by fitting an analytic function to the ab initio potential energy values computed using coupled cluster singles and doubles with perturbative triples [CCSD(T)] method and Dunning's augmented correlation consistent polarized valence triple zeta basis set. Using this potential energy surface, time-dependent quantum mechanical wave packet calculations were carried out to calculate the reaction probabilities (P(R)) for the exchange reaction H( )+H(2)(v, j)-->H(2)+H(-), for different initial vibrational (v) and rotational (j) states of H(2), for total angular momentum equal to zero. With increase in v, the number of oscillations in the P(R)(E) plot increases and the oscillations become more pronounced. While P(R) increases with increase in rotational excitation from j=0 to 1, it decreases with further increase in j to 2 over a wide range of energies. In addition, rotational excitation quenches the oscillations in P(R)(E) plots. PMID- 15538854 TI - Nonadiabatic energy transfer studies of O((1)D)+N(2)(X (1)Sigma(g) (+))- >O((3)P)+N(2)(X (1)Sigma(g) (+)) by time-dependent wave packet. AB - Three-dimensional time-dependent quantum calculations have been performed on two/three coupled potential surfaces, including the singlet surface 1 (1)A(') and two triplet surfaces 1 (3)A(') and 1 (3)A("), for the electronic quenching process of O((1)D)+N(2)(X (1)Sigma(g) (+))-->O((3)P)+N(2)(X (1)Sigma(g) (+)). An extended split-operator scheme was employed to study this nonadiabatic process. Two types of singlet surface 1 (1)A('), namely, double many body expansion (DMBE2) were used in the calculations, along with spin-orbit couplings of Nakamura-Kato and with a constant value of 80 cm(-1). All the calculated probabilities are resonance dominated, with a general decreasing trend within the investigated collision energy range. The probability involving three potential energy surfaces is approximately two times as high as that on two potential energy surfaces. At low collision energies, the calculations on the ZPM2 surface produced much larger probability than that on the DMBE2 surface, but the difference was diminishing as the collision energy became high. The behavior of the probability on DMBE2/ZPM2 surfaces at low energies indicates that the ZPM2 surface dominates over the DMBE2 surface in the description of the process. However, the DMBE2 surface has been modified by removing the unreasonable barrier. The estimated quenching cross sections both on the ZPM2 surface and on the modified DMBE2 surface in the three-coupled-surface calculations agree with the experimental measurement. Also, a rather insensitive characteristic of the probability relative to the analytical function form of spin-orbit coupling is revealed. PMID- 15538855 TI - The ability of silylenes to bind excess electrons: electron affinities of SiX(2), and SiXY species (X,Y=H,CH(3),SiH(3),F,Cl,Br). AB - Recently, Ishida and co-workers have isolated silylene radical anions via the one electron reduction of isolable cyclic dialkylsilylenes, discovering these corresponding radical anions to be relatively stable at low temperatures. Herein we report theoretical predictions of the adiabatic electron affinities (AEA), vertical electron affinities, and vertical detachment energies of a series of methyl, silyl, and halosubstituted silylene compounds. This research utilizes the carefully calibrated DZP++ basis with the combination of the popular nonhybrid and hybrid DFT functionals, BLYP, B3LYP, and BHHLYP. The level of theory employed and the ensemble of species under study confirm the ability of silylenes to bind excess electrons with Si(SiH(3))(2) being the most effective, having a predicted AEA of 1.95 eV. While it is known that methyl substituents have a diminishing effect on the computed electron affinities (EAs), it is shown that fluorine shows an analogous negative effect. Similarly, previous suggestions that Si(CH(3))(2) will not bind an electron appear incorrect, with EA[Si(CH(3))(2)] predicted here to be 0.46 eV. PMID- 15538856 TI - Photodissociation of CCH: classical trajectory calculations involving seven electronic states. AB - The photodissociation dynamics of ethynyl radical, C(2)H, involving seven electronic states is studied by classical trajectory calculations. Initial values of the trajectories are selected based on relative absorption intensities calculated by Mebel et al. The energies and the derivatives are interpolated by three-dimensional cubic spline interpolator using an extended data pool. Mean square errors and standard deviations in interpolation of energies for 450 data points are found to be in the range 3.1 x 10(-6)-1.4 x 10(-5) and 1.7 x 10(-3) 3.8 x 10(-3) hartrees, respectively. The photofragments of C(2) and H are produced mainly in the X (1)Sigma(g) (+), a (3)Pi(u), b (3)Sigma(g) (-), c (3)Sigma(u) (+), A (1)Pi(u), B (1)Delta(g) electronic states of C(2) as product. The avoided crossings do not appear to be in the main dissociation pathways. The internal distributions are in good accord with the experimental results where comparison is possible, suggesting that the fragmentation mechanism of C(2)H(2) into C(2) and H is a two step process involving C(2)H radical as an intermediate with a life time long enough to allow complete collection of the phase space in the experiments. PMID- 15538857 TI - Application of direct potential fitting to line position data for the X (1)Sigma(+) and A (1)Sigma(+) states of LiH. AB - A collection of 9089 spectroscopic LiH line positions, of widely varying precision, which sample 84.9% and 98.6% of the A and X state well depths, respectively, have been employed in a direct least-squares fit of the effective potential energy and Born-Oppenheimer breakdown functions for the two states. For the four isotopomers (6)LiH, (7)LiH, (6)LiD, and (7)LiD, the data comprise both pure rotational and vibration-rotational transitions within the ground state, as well as rotationally resolved transitions in the A-X system. Despite the unusual shape and associated anomalous properties of the A state potential, no special features or considerations were required in the direct potential fitting approach. The reduced standard deviation of the fit was close to unity, indicating that the quantum mechanical eigenvalues calculated from the fully analytical functions of the Hamiltonians of the two states, which are characterized by a total of only 53 fitted parameters, represent the line positions, on average, to within the estimated uncertainties. A quantum mechanical calculation of the molecular constants G(nu), B(nu), D(nu), H(nu), L(nu), M(nu), N(nu), and O(nu) from the fitted potential for the A state of (7)LiH confirms that the usual polynomial expansion in J(J+1) is an unsatisfactory representation for the rotational terms of the lowest vibrational levels. PMID- 15538858 TI - On the ultraviolet photodissociation of H(2)Te. AB - The photodissociation of H(2)Te through excitation in the first absorption band is investigated by means of multireference spin-orbit configuration interaction (CI) calculations. Bending potentials for low-lying electronic states of H(2)Te are obtained in C(2v) symmetry for Te-H distances fixed at the ground state equilibrium value of 3.14a(0), as well as for the minimum energy path constrained to R(1)=R(2). Asymmetric cuts of potential energy surfaces for excited states (at R(1)=3.14a(0) and theta;=90.3 degrees ) are obtained for the first time. It is shown that vibrational structure in the 380-400 nm region of the long wavelength absorption tail is due to transitions to 3A('), which has a shallow minimum at large HTe-H separations. Transitions to this state are polarized in the molecular plane, and this state converges to the excited TeH((2)Pi(1/2))+H((2)S) limit. These theoretical data are in accord with the selectivity toward TeH((2)Pi(1/2)) relative to TeH((2)Pi(3/2)) that has been found experimentally for 355 nm H(2)Te photodissociation. The calculated 3A(')<--XA(') transition dipole moment increases rapidly with HTe-H distance; this explains the observation of 3A(') vibrational structure for low vibrational levels, despite unfavorable Franck Condon factors. According to the calculated vertical energies and transition moment data, the maximum in the first absorption band at approximately 245 nm is caused by excitation to 4A("), which has predominantly 2(1)A(") ((1)B(1) in C(2v) symmetry) character. PMID- 15538859 TI - Fine structure of the (S(1)<--S(0)) band origins of phthalocyanine molecules in helium droplets. AB - The laser-induced fluorescence (LIF) excitation spectra of free base phthalocyanine (Pc), Mg-Pc, and Zn-Pc molecules in superfluid helium droplets at T=0.38 K have been studied. The spectra reveal the rich vibronic structure of the S(1)<--S(0) electronic transitions. The band origins of the transitions consist of zero phonon lines accompanied by phonon wings, which originate from simultaneous electronic excitation of the molecule and excitation of the collective modes of the helium surrounding it. The phonon wings have discrete structures suggesting localization of some helium atoms in the neighborhood of the molecules. Zero phonon lines of Mg-Pc and Zn-Pc molecules are split into three components, which are separated by 0.2-0.4 cm(-1). Possible mechanism of splitting involves static or dynamic Jahn-Teller interaction of metal phthalocyanine molecules in the twofold degenerate S(1)((1)E(u)) state with the helium shell. PMID- 15538860 TI - Collision energy transfer in collision of NH(4) (+)(NH(3))(n-1) (n=3-9) with ND(3). AB - An incorporation of ND(3) into protonated ammonia cluster ions NH(4)(+)(NH(3))(n 1) (n=3-9), together with a dissociation of the cluster ions, was observed in the collision of the cluster with ND(3) at collision energies ranging from 0.04 to 1.4 eV in the center-of-mass frame. The branching fractions of the cluster ion species produced in the reactions were obtained as a function of the collision energy. The branching fractions of the incorporation products were successfully explained in terms of the Rice-Ramsperger-Kassel (RRK) theory at collision energies lower than the binding energy of the cluster ion. In addition, the internal energy distributions of the parent cluster ions were determined, and found to be in good agreement with those predicted using the evaporative ensemble model. In incorporations at collision energies lower than the binding energy of the cluster ion, all of the collision energy was transferred to the internal energy of the cluster ions; subsequently, an evaporation of ammonia molecules occurred in an equilibrium process after a complete energy redistribution in the clusters. In contrast, at collision energies higher than the binding energy of the cluster ion, a release of an ammonia molecule from the incorporation products occurred in a nonequilibrium process. The transition from the complex mode to the direct mode in the incorporation was observed at collision energies approximately equal to the binding energy. On the other hand, the collision energy dependence of the cross sections for the dissociation and for a nonreactive collision were estimated by a RRK simulation in which the collision energy transfer was interpreted by using the classical hard-sphere collision model. A relationship between reactivity and reaction modes in the collision of NH(4)(+)(NH(3))(4) with ND(3) is discussed via a comparison of the experimental results with the RRK simulation. PMID- 15538861 TI - Appearance of bulk properties in small tungsten oxide clusters. AB - Contrary to the conventional understanding that atomic clusters usually differ in properties and structure from the bulk constituents of which they are comprised, we show that even a dimer of tungsten oxide (WO(3))(2) possesses bulklike features and the geometry of a small cluster containing only 4 tungsten and 12 oxygen atoms bears the hallmarks of crystalline tungsten oxide, WO(3). This observation, based on a synergistic approach involving mass distributions under quasisteady state conditions, photoelectron spectroscopy, and first principles molecular orbital theory, not only illustrates the existence of a class of strongly covalent or ionic materials whose embryonic forms are tiny clusters but also lends the possibility that a fundamental understanding of complex processes such as catalytic reactions on surfaces may be achieved on an atomic scale with clusters as model systems. PMID- 15538862 TI - Differential and total (e,2e) cross sections of simple polyatomic molecules. AB - In this paper, we present a theoretical approach to calculate differential and total ionization cross sections of polyatomic molecules by fast electron impact. More exactly, we have studied the ionization of ammonia (NH(3)) and methane (CH(4)) molecules, and previous results concerning the H(2)O molecule ionization are reported for comparison. The calculations are performed in the distorted wave Born approximation without exchange by employing the independent electron model. The molecular target wave functions are described by linear combinations of atomic orbitals. To describe the interaction between the inactive target electrons and the slow ejected electron, we have introduced a distortion via an effective potential calculated for each molecular orbital. The present theoretical calculations agree well with a large set of existing experimental data in terms of multiple differential and total cross sections. PMID- 15538863 TI - Near-infrared laser spectroscopy of NiI. AB - Laser-induced fluorescence spectrum of NiI in the near infrared region of 714-770 nm has been recorded. Seven bands belonging to three electronic transition systems were observed and analyzed: the (0,0), (1,0), and (2,0) bands of [13.3] (2)Sigma(+)-A (2)Pi(3/2) system; the (1,1) and (0,1) bands of [13.9] (2)Pi(3/2)-X (2)Delta(5/2) system; and the (0,0) and (1,0) bands of [13.9] (2)Pi(3/2)-A (2)Pi(3/2) system. Spectra of isotopic molecules confirmed the vibrational quantum number assignment of the observed bands. Least-squares fit of rotationally resolved transition lines yielded accurate molecular constants for the v=0-2 levels of the [13.3] (2)Sigma(+) state, the v=0 level of the A (2)Pi(3/2), and the v=1 level of the X (2)Delta(5/2) state. The vibrational separation, DeltaG(1/2), of the ground state was measured to be 276.674 cm(-1). With the observation of the [13.9] (2)Pi(3/2)-A (2)Pi(3/2) and [13.9] (2)Pi(3/2) X (2)Delta(5/2) transitions, we accurately determined the energy separation between the A (2)Pi(3/2) and the X (2)Delta(5/2) to be 163.847 cm(-1). This confirms that the order of the A (2)Pi(3/2) and X (2)Delta(5/2) states in NiI is reversed when compared with other nickel monohalides. PMID- 15538864 TI - Ultrafast excited-state dynamics in photochromic N-salicylideneaniline studied by femtosecond time-resolved REMPI spectroscopy. AB - Ultrafast processes in photoexcited N-salicylideneaniline have been investigated with femtosecond time-resolved resonance-enhanced multiphoton ionization spectroscopy. The ion signals via the S(1)(n,pi( *)) state of the enol form as well as the proton-transferred cis-keto form emerge within a few hundred femtoseconds after photoexcitation to the first S(1)(pi,pi( *)) state of the enol form. This reveals that two ultrafast processes, excited-state intramolecular proton transfer (ESIPT) reaction and an internal conversion (IC) to the S(1)(n,pi( *)) state, occur on a time scale less than a few hundred femtoseconds from the S(1)(pi,pi( *)) state of the enol form. The rise time of the transient corresponding to the production of the proton-transferred cis-keto form is within 750 fs when near the red edge of the absorption is excited, indicating that the ESIPT reaction occurs within 750 fs. The decay time of the S(1)(pi,pi( *)) state of the cis-keto form is 8.9 ps by exciting the enol form at 370 nm, but it dramatically decreases to be 1.5-1.6 ps for the excitation at 365-320 nm. The decrease in the decay time has been attributed to the opening of an efficient nonradiative channel; an IC from S(1)(pi,pi( *)) to S(1)(n,pi( *)) of the cis keto form promotes the production of the trans-keto form as the final photochromic products. The two IC processes may provide opposite effect on the quantum yield of photochromic products: IC in the enol form may substantially reduce the quantum yield, but IC in the cis-keto form increase it. PMID- 15538865 TI - Vibrational relaxation of oxygen in an argon cage. AB - The vibrational relaxation of oxygen embedded in an argon cage through vibrational to local translation, rotation, and argon phonon modes has been studied using semiclassical procedures. The collision model is based on the trapped molecule undergoing the restricted motions (local translation and hindered rotation) in a cage formed by its twelve nearest argon neighbors in a face-centered-cubic structure. At 85 K in the liquid argon temperature range, the deexcitation probability of O(2)(v=1) is 5.8 x 10(-12) and the relaxation rate constant with the collision frequency from local translation is 23 s(-1). The rate constant decreases to 5.1 s(-1) at 50 K and to 0.016 s(-1) at 10 K in the solid argon temperature range. Transfer of the vibrational energy to local translation, rotation (both hindered and free), and argon phonon modes is the relaxation pathway for the trapped oxygen molecule. PMID- 15538866 TI - A study of the correlation effects upon the modelization of the double exchange phenomenon. AB - A previous work by the authors has shown that the usual spin Hamiltonian used to model the magnetic spectra of mixed valence compounds was not sufficient to reproduce the magnetic spectrum of the molecule [Fe(2)(OH)(3)(NH(3))(6)](2+). In the present paper, the spin Hamiltonian is confronted to correlated ab initio calculations. The discrepancy between this Hamiltonian and the calculations is investigated and explained. It is pointed out that the multiconfigurational nature of the transition metal is responsible for this discrepancy. However, we show that this effect can easily be treated at the complete active space self consistent field (CASSCF) level and that no further correlation treatment is needed. The spin Hamiltonian, which reproduces very well the minimal CASSCF results, could not be improved to recover the multireference effects. PMID- 15538867 TI - The Cotton-Mouton effect of neon and argon: a benchmark study using highly correlated coupled cluster wave functions. AB - The Cotton-Mouton effect (magnetic field induced linear birefringence) has been studied for neon and argon using state-of-the-art coupled cluster techniques. The coupled cluster singles, doubles and triples (CCSDT) approach has been used to obtain static benchmark results and the CC3 model with an approximate treatment of triple excitations to obtain frequency-dependent results. In the case of neon the effect of excitations beyond triples has also been estimated via coupled cluster calculations including quadruple excitations (CCSDTQ), pentuple excitations (CCSDTQP), etc. up to the full configuration-interaction level. The results obtained for the anisotropy of the hypermagnetizability Deltaeta(omega), the molecular property that determines the magnetic field induced birefringence of spherically symmetric systems, are Deltaeta=2.89 a.u. for neon and Deltaeta=24.7 a.u. for argon, with a negligible effect of frequency dispersion. For neon we could estimate an absolute error on Deltaeta of 0.1 a.u. The accuracy of these results surpasses that of recently reported experimental data. PMID- 15538868 TI - Direct ab initio dynamics study on the rate constants and kinetics isotope effects of CH(3)O+H-->CH(2)O+H(2) reaction. AB - We present a direct ab initio dynamics study of thermal rate constants of the hydrogen abstraction reaction of CH(3)O+H-->CH(2)O+H(2). The unrestricted Becke's half-and-half hybrid functional using the Lee-Yang-Parr correlation functional with Dunning's correlation consistent polarized valence double-zeta basis set, the unrestricted quadratic configuration interaction calculation including single and double substitutions with Dunning's correlation consistent polarized valence double-zeta basis set, and the unrestricted quadratic configuration interaction calculation including single and double substitutions with a triples contribution with Dunning's correlation consistent polarized valence triple-zeta basis set methods were employed to optimize the structures and to calculate frequencies for all stationary points. Minimum energy paths were obtained by the unrestricted Becke's half-and-half hybrid functional using the Lee-Yang-Parr correlation functional and the unrestricted quadratic configuration interaction calculation including single and double substitutions with the same Dunning's correlation consistent polarized valence double-zeta basis set levels of theory. No barrier is found at the unrestricted Becke's half-and-half hybrid functional using the Lee-Yang-Parr correlation functional with Dunning's correlation consistent polarized valence double-zeta basis set level of theory in contrast to a small barrier of 1.43 kcal mol(-1) at the unrestricted quadratic configuration interaction calculation including single and double substitutions with Dunning's correlation consistent polarized valence double-zeta basis set level of theory. In particular, the barrier vanishes as the energies along the minimum energy path MEP are refined at the unrestricted quadratic configuration interaction calculation including single and double substitutions with a triples contribution with Dunning's correlation consistent polarized valence triple-zeta basis set level of theory. Smaller barriers of 0.47 and 0.17 kcal mol(-1) were obtained at the unrestricted quadratic configuration interaction calculation including single and double substitutions with a triples contribution with Dunning's correlation consistent polarized valence triple-zeta basis set and the unrestricted quadratic configuration interaction calculation including single and double substitutions with a triples contribution with Dunning's correlation consistent polarized valence triple-zeta basis set based on the geometries at the unrestricted quadratic configuration interaction calculation including single and double substitutions with Dunning's correlation consistent polarized valence triple-zeta basis set levels of theory, respectively. The forward rate constants are evaluated with the canonical variational transition state theory in the temperature range of 300-2500 K. The calculated forward rate constants at the unrestricted quadratic configuration interaction calculation including single and double substitutions with a triples contribution with Dunning's correlation consistent polarized valence triple-zeta basis set based on the geometries at the unrestricted quadratic configuration interaction calculation including single and double substitutions with Dunning's correlation consistent polarized valence double-zeta basis set level of theory are in good agreement with the available experimental data. The kinetic isotope effects are estimated. PMID- 15538869 TI - Ion-molecule rate constants and branching ratios for the reaction of N(3) (+)+O(2) from 120 to 1400 K. AB - The kinetics of the reaction of N(3) (+) with O(2) has been studied from 120 to 1400 K using both a selected ion flow tube and high-temperature flowing afterglow. The rate constant decreases from 120 K to approximately 1200 K and then increases slightly up to the maximum temperature studied, 1400 K. The rate constant compares well to most of the previous measurements in the overlapping temperature range. Comparing the results to drift tube data shows that there is not a large difference between increasing the translational energy available for reaction and increasing the internal energy of the reactants over much of the range, i.e., all types of energies drive the reactivity equally. The reaction produces both NO(+) and NO(2) (+), the latter of which is shown to be the higher energy NOO(+) linear isomer. The ratio of NOO(+) to NO(+) decreases from a value of over 2 at 120 K to less than 0.01 at 1400 K because of dissociation of NOO(+) at the higher temperatures. This ratio decreases exponentially with increasing temperature. High-level theoretical calculations have also been performed to compliment the data. Calculations using multi-reference configuration interaction theory at the MRCISD(Q)/cc-pVTZ level of theory show that singlet NOO(+) is linear and is 4.5 eV higher in energy than ONO(+). A barrier of 0.9 eV prevents dissociation into NO(+) and O((1)D); however, a crossing to a triplet surface connects to NO(+) and O((3)P) products. A singlet and a triplet potential energy surface leading to products have been determined using coupled cluster theory at the CCSD(T)/aug-cc-pVQZ level on structures optimized at the Becke3-Lee, Yang, and Parr (B3LYP)/aug-cc-pVTZ level of theory. The experimental results and reaction mechanism are evaluated using these surfaces. PMID- 15538870 TI - Pendular-state spectroscopy of the S(1)-S(0) electronic transition of 9 cyanoanthracene. AB - Fluorescence excitation spectra of the S(1)-S(0) origin band of 9-cyanoanthracene have been observed under a uniform electric field up to 200 kV/cm to explore pendular-state spectrum of an asymmetric-top molecule close to the strong field limit. The observed spectra exhibit distinct evolution of the band contour as a function of the applied electric field, which are much different from each other for different excitation configurations. An approximate method suitable for spectrum simulations of large asymmetric-top molecules in a pendular condition is developed for the analysis of the experimental results. The comparison of the observed and simulated spectra shows that the spectra are well ascribed in terms of the pendular-state selection rules, which have recently been derived from theoretical consideration of the pendular-limit representation of energy levels and spectra. PMID- 15538871 TI - Protonation study of peroxynitric acid and peroxynitrous acid. AB - The equilibrium structures and harmonic vibrational frequencies of peroxynitric acid (HOONO(2)) and seven structures of protonated peroxynitric acid, along with peroxynitrous acid (HOONO) and its 12 protonated peroxynitrous acid structures, have been investigated using several ab initio and density functional methods. The ab initio methods include second-order Moller-Plesset perturbation theory, quadratic configuration interaction, including single and double excitations theory (QCISD), and the QCISD(T) methods, which incorporate a perturbational estimate of the effects of connected triple excitation. The Becke three-parameter hybrid functional combined with Lee, Yang, and Parr correlation function is the density functional method used. The lowest energy form of protonated peroxynitric acid is a complex between H(2)O(2) and NO(+) rather than between H(2)O and NO(2) (+). For peroxynitrous acid, a complex between H(2)O(2) and NO(2) (+) is found to be the lowest energy structure. The ab initio proton affinity (PA) of HOONO and HOONO(2) is predicted to be 182.1 and 175.1 kcal mol(-1), respectively, at the QCISD(T)/6-311++G(3df,3pd) level of theory. The results are contrasted with an earlier study on nitrous acid, and is shown that peroxynitric acid and peroxynitrous acid have a smaller PA than nitrous acid. PMID- 15538872 TI - Nucleation rates of water and heavy water using equations of state. AB - The original formula of Gibbs for the reversible work of critical nucleus formation is evaluated in three approximate ways for ordinary and heavy water. The least approximate way employs an equation of state to evaluate the pressure difference between the new and old phases. This form of the theory yields a temperature dependence for the nucleation rate close to that observed experimentally. This is a substantial improvement over the most commonly used (and most approximate) form of classical theory. PMID- 15538873 TI - Electric conductivities of 1:1 electrolytes in liquid methanol along the liquid vapor coexistence curve up to the critical temperature. II. KBr and KI solutions. AB - The molar conductivities Lambda of KBr and KI in dilute methanol solutions were measured along the liquid-vapor coexistence curve up to the critical temperature (240 degrees C). The concentration dependence of Lambda in each condition was analyzed by the Fuoss-Chen-Justice equation to obtain the limiting molar conductivities and the molar association constants. Using the present data together with the literature ones, the validity of the Hubbard-Onsager (HO) dielectric friction theory based on the sphere-in-continuum model was examined for the translational friction coefficients zeta of the halide ions (the Cl(-), Br(-), and I(-) ions) in methanol in the density range of 2.989rho(c)> or =rho> or =1.506rho(c), where rho(c)=0.2756 g cm(-3) is the critical density of methanol. For all the halide ions studied, the friction coefficient decreased with decreasing density at rho>2.0rho(c), while the nonviscous contribution Deltazeta/zeta increased; Deltazeta was defined as the difference between zeta and the friction coefficient estimated by the Stokes law. The density dependence of zeta and Deltazeta/zeta were well reproduced by the HO theory at rho>2.0rho(c). The HO theory also explained the ion-size dependence of Deltazeta/zeta which decreased with ion-size at rho>2.0rho(c). At rho<2.0rho(c), on the other hand, the HO theory could not explain the density and the ion-size dependences of zeta and Deltazeta/zeta. These results indicated that the application limit of the HO theory lied about rho=2.0rho(c) which is the same as the application limit observed for the alkali metal ions. The present results were also compared with the results in subcritical aqueous solutions. PMID- 15538874 TI - Transport coefficients of the Lennard-Jones model fluid. II Self-diffusion. AB - In an extensive computer simulation study, the transport coefficients of the Lennard-Jones model fluid were determined with high accuracy from equilibrium molecular-dynamics simulations. In the frame of time-correlation function theory, the generalized Einstein relations were employed to evaluate the transport coefficients. This second of a series of four papers presents the results for the self-diffusion coefficient, and discusses and interprets the behavior of this transport coefficient in the fluid region of the phase diagram. The uncertainty of the self-diffusion data is estimated to be 1% in the gas region and 0.5% at high-density liquid states. With the very accurate data, even fine details in the shape of the self-diffusion isotherms are resolved, and the previously little investigated behavior of the self-diffusion coefficient at low-density gaseous states is analyzed in detail. Finally, aspects of the mass transport mechanisms on the molecular scale are explored by an analysis of the velocity autocorrelation functions. PMID- 15538875 TI - Separation of metal ions in nitrate solution by ultrasonic atomization. AB - In the ultrasonic atomization of metal nitrate solutions, the molar ratio of metal ions is changed between solution and mist. Small molar metal ions tend to be transferred to mist by ultrasonic wave acceleration, while large molar ions tend to remain in solution. As a result, metal ions can be separated by ultrasonic atomization. We show experimental data and propose a conceptual mechanism for the ultrasonic separation of metal ions. PMID- 15538876 TI - Calculation of the aqueous solvation energy and entropy, as well as free energy, of simple polar solutes. AB - With the advent of more powerful computers, the question of calculating thermodynamic quantities, such as the energy and the entropy, in solute-solvent systems is revisited. The calculation of these thermodynamic quantitites was limited in the past by their slow convergence relative to the free energy. Using molecular dynamics simulations, the energy, entropy, and free energy of solvation of NMA and CH(3)NH(2), as well as their relative values, have been determined. Three different methods (the thermodynamic perturbation method, the thermodynamic integration method, and a finite-difference method) are compared. The finite difference method gives the best results and accurate values for the entropy and energy were obtained using a reasonable amount to computer time. The results suggest that a meaningful thermodynamic description of biomolecular processes can be realized with present methods and the available computer time. PMID- 15538877 TI - Charge-on-spring polarizable water models revisited: from water clusters to liquid water to ice. AB - The properties of two improved versions of charge-on-spring (COS) polarizable water models (COS/G2 and COS/G3) that explicitly include nonadditive polarization effects are reported. In COS models, the polarization is represented via a self consistently induced dipole moment consisting of a pair of separated charges. A previous polarizable water model (COS/B2), upon which the improved versions are based, was developed by Yu, Hansson, and van Gunsteren. To improve the COS/B2 model, which overestimated the dielectric permittivity, one additional virtual atomic site was used to reproduce the water monomer quadrupole moments besides the water monomer dipole moment in the gas phase. The molecular polarizability, residing on the virtual atomic site, and Lennard-Jones parameters for oxygen oxygen interactions were varied to reproduce the experimental values for the heat of vaporization and the density of liquid water at room temperature and pressure. The improved models were used to study the properties of liquid water at various thermodynamic states as well as gaseous water clusters and ice. Overall, good agreement is obtained between simulated properties and those derived from experiments and ab initio calculations. The COS/G2 and COS/G3 models may serve as simple, classical, rigid, polarizable water models for the study of organic solutes and biopolymers. Due to its simplicity, COS type of polarization can straightforwardly be used to introduce explicit polarization into (bio)molecular force fields. PMID- 15538878 TI - A molecular dynamics simulation of the melting points and glass transition temperatures of myo- and neo-inositol. AB - The heat of sublimation, density, melting point, and glass transition temperature are calculated for myo- and neo-inositol, using the condensed-phase optimized molecular potentials for atomistic simulation studies (COMPASS) force field and molecular dynamics techniques. Our results show that the calculated heats of sublimation and density are very close to the experimental values for both compounds. Furthermore, our simulated melting temperatures for myo- and neo inositol also compare very well to the experimentally obtained data. The glass transition temperatures for myo- and neo-inositol have been calculated to be ca. 494 K and ca. 518 K, respectively, and the shape of the volume versus temperature plots produced are typical for a glass transition. As a result, it is our view that the COMPASS force field suitably describes these two compounds in molecular simulations and that molecular dynamics techniques, combined with this force field, can be used to simulate the melt and glass transitions for such molecules. PMID- 15538879 TI - Molecular simulation study of effect of molecular association on vapor-liquid interfacial properties. AB - Vapor-liquid interfacial properties of square-well associating fluids are studied via transition-matrix Monte Carlo simulation. Results for one-site and two-site association models are presented. Coexistence properties, surface tension, cluster distribution, density profile, and orientation profile are presented. Molecular association affects the interfacial properties and cluster fractions more than it affects the bulk densities. We observe that the surface tension exhibits a maximum with respect to association strength. This behavior is in agreement with the recent study of Peery and Evans for one site system using a square-gradient approach. PMID- 15538880 TI - Molecular dynamics averaging of Xe chemical shifts in liquids. AB - The Xe nuclear magnetic resonance chemical shift differences that afford the discrimination between various biological environments are of current interest for biosensor applications and medical diagnostic purposes. In many such environments the Xe signal appears close to that in water. We calculate average Xe chemical shifts (relative to the free Xe atom) in solution in eleven liquids: water, isobutane, perfluoro-isobutane, n-butane, n-pentane, neopentane, perfluoroneopentane, n-hexane, n-octane, n-perfluorooctane, and perfluorooctyl bromide. The latter is a liquid used for intravenous Xe delivery. We calculate quantum mechanically the Xe shielding response in Xe-molecule van der Waals complexes, from which calculations we develop Xe (atomic site) interpolating functions that reproduce the ab initio Xe shielding response in the complex. By assuming additivity, these Xe-site shielding functions can be used to calculate the shielding for any configuration of such molecules around Xe. The averaging over configurations is done via molecular dynamics (MD). The simulations were carried out using a MD technique that one of us had developed previously for the simulation of Henry's constants of gases dissolved in liquids. It is based on separating a gaseous compartment in the MD system from the solvent using a semipermeable membrane that is permeable only to the gas molecules. We reproduce the experimental trends in the Xe chemical shifts in n-alkanes with increasing number of carbons and the large chemical shift difference between Xe in water and in perfluorooctyl bromide. We also reproduce the trend for a given solvent of decreasing Xe chemical shift with increasing temperature. We predict chemical shift differences between Xe in alkanes vs their perfluoro counterparts. PMID- 15538881 TI - Bimodal proton transfer in acid-base reactions in water. AB - We investigate one of the fundamental reactions in solutions, the neutralization of an acid by a base. We use a photoacid, 8-hydroxy-1,3,6-trisulfonate-pyrene (HPTS; pyranine), which upon photoexcitation reacts with acetate under transfer of a deuteron (solvent: deuterated water). We analyze in detail the resulting bimodal reaction dynamics between the photoacid and the base, the first report on which was recently published. We have ascribed the bimodal proton-transfer dynamics to contributions from preformed hydrogen bonding complexes and from initially uncomplexed acid and base. We report on the observation of an additional (6 ps)(-1) contribution to the reaction rate constant. As before, we analyze the slower part of the reaction within the framework of the diffusion model and the fastest part by a static, sub-150 fs reaction rate. Adding the second static term considerably improves the overall modeling of the experimental results. It also allows to connect experimentally the diffusion controlled bimolecular reaction models as defined by Eigen-Weller and by Collins-Kimball. Our findings are in agreement with a three-stage mechanism for liquid phase intermolecular proton transfer: mutual diffusion of acid and base to form a "loose" encounter complex, followed by reorganization of the solvent shells and by "tightening" of the acid-base encounter complex. These rearrangements last a few picoseconds and enable a prompt proton transfer along the reaction coordinate, which occurs faster than our time resolution of 150 fs. Alternative models for the explanation of the slower "on-contact" reaction time of the loose encounter complex in terms of proton transmission through a von Grotthuss mechanism are also discussed. PMID- 15538882 TI - Ion dynamics in mixed alkali cadmium fluoride glasses. AB - We have investigated ion dynamics in cadmium fluoride nonoxide glasses, in the frequency range from 10 Hz to 2 MHz and in the temperature range from room temperature to just below the glass transition temperature. We have established the existence of mixed alkali effect for these glasses from different aspects of relaxation dynamics. We have observed lower dimensionality of the conduction pathways in mixed alkali cadmium fluoride glasses compared to that in the single alkali glasses. Although, both n and beta represent the interaction between the charge carriers, we have not found any theoretical correlation between them. Using different scaling approaches we have shown that the relaxation dynamics in mixed alkali cadmium fluoride glasses is independent of temperatures. PMID- 15538883 TI - Synthesis of silver molybdate clusters driven by laser-annealing. AB - The synthesis of silver rich molybdate clusters is achieved by laser induced chemical reaction of coadsorbed MoO(3) and O(2) molecules on free silver clusters. The reactants MoO(3) and/or O(2) molecules condensed at low temperature (77 K-175 K) on free silver clusters. Then, the silver clusters together with their adsorbed molecules are flashed either ionized with a discharge or ionized and heated by a laser. Then they are cooled down by evaporation. The synthesized chemical compounds are analyzed by a high-resolution time-of-flight mass spectrometer. If only one type of reactant is adsorbed on the cluster, only one oxide molecule is stabilized on the metallic core after the heating and cooling cycle. On the contrary, the coadsorption of the two types of molecules MoO(3) and O(2) on Ag(n) (+), at 77 K, leads to complex aggregates that transform, after laser heating, into a molybdate rich metal clusters. These synthesized species cool down by evaporating silver atoms showing evidence of a binary oxide that is more stable than the metallic core. Moreover we demonstrate that for small size molybdate clusters, a stoichiometric composition may differ from the bulk one. PMID- 15538884 TI - Vapor-liquid transitions of dipolar fluids in disordered porous media: performance of angle-averaged potentials. AB - Using replica integral equations in the reference hypernetted-chain (RHNC) approximation we calculate vapor-liquid spinodals, chemical potentials, and compressibilities of fluids with angle-averaged dipolar interactions adsorbed to various disordered porous media. Comparison with previous RHNC results for systems with true angle-dependent Stockmayer (dipolar plus Lennard-Jones) interactions indicate that, for a dilute hard sphere matrix, the angle-averaged fluid-fluid (ff) potential is a reasonable alternative for reduced fluid dipole moments m( *2)=mu(2)/(epsilon(0)sigma(3))< or =2.0. This range is comparable to that estimated in bulk fluids, for which RHNC results are presented as well. Finally, results for weakly polar matrices suggest that angle-averaged fluid matrix (fm) interactions can reproduce main features observed for true dipolar (fm) interactions such as the shift of the vapor-liquid spinodals towards lower temperatures and higher densities. However, the effective attraction induced by dipolar (fm) interaction is underestimated rather than overestimated as in the case of angle-averaged ff interactions. PMID- 15538885 TI - Interfacial stability of electrodeposition of cuprous oxide films. AB - Experiments on deposition of Cu(2)O films from basic copper sulfate solution show that copper also deposits. At low, but basic values of pH only copper deposits and at high pH only cuprous oxide deposits. In the intermediate range where both compete the system shows oscillations at "constant current." Linear stability analysis has been conducted for such an electrochemical cell to show that oscillations can take place in the parameter space identified in the experiments. The results are keeping with most of the experimental observations, which are many, but not with all. The physical mechanisms behind the oscillations are explained in terms of competing reactions. PMID- 15538886 TI - The effect of surface roughness on the adhesion of solid surfaces for systems with and without liquid lubricant. AB - We present molecular dynamics results for the interaction between two solid elastic walls during pull-off for systems with and without octane (C(8)H(18)) lubricant. We used two types of substrate--flat and corrugated--and varied the lubricant coverage from approximately 1/8 to approximately 4 ML (monolayers) of octane. For the flat substrate without lubricant the maximum adhesion was found to be approximately three times larger than for the system with the corrugated substrate. As a function of the octane coverage (for the corrugated substrate) the pull-off force first increases as the coverage increases from 0 to approximately 1 ML, and then decreases as the coverage is increased beyond monolayer coverage. It is shown that at low octane coverage, the octane molecules located in the substrate corrugation wells during squeezing are pulled out of the wells during pull-off, forming a network of nanocapillary bridges around the substrate nanoasperities, thus increasing the adhesion between two surfaces. For greater lubricant coverages a single capillary bridge is formed. The adhesion force saturates for lubricant coverages greater than 3 ML. For the flat substrate, during pull-off we observe discontinuous, thermally activated changes in the number n of lubricant layers (n-1-->n layering transitions), whereas for the corrugated substrate these transitions are "averaged" by the substrate surface roughness. PMID- 15538887 TI - Comparative molecular dynamics study of ether- and ester-linked phospholipid bilayers. AB - The lipid membranes found in archaea have high bilayer stability and low permeability. The molecular structure of their constituent lipids is characterized by ether-linked, branched hydrophobic chains, whereas the conventional lipids obtained from eukaryotic or eubacterial sources have ester linked straight chains. In order to elucidate the influence of the ether linkage, instead of an ester one, on the physical properties of the lipid bilayers, we have carried out comparative 10 ns molecular dynamics simulations of diphytanyl phosphatidylcholine (ether-DPhPC) and diphytanoyl phosphatidylcholine (ester DPhPC) bilayers in water, respectively. We analyze bilayer structures, hydration of the lipids, membrane dipole potentials, and free energy profiles of water and oxygen across the bilayers. We observe that the membrane dipole potential for the ether-DPhPC bilayer, which arises mainly from the ether linkage, is about half of that of the ester-DPhPC. The calculated free energy barrier for a water molecule in the ether-DPhPC bilayer system is slightly higher than that in the ester-DPhPC counterpart, which is in accord with experimental data. PMID- 15538888 TI - Influence of molecular adsorption on the dielectric properties of a single wall nanotube: a model sensor. AB - Recent measurements of the resonance frequency of a copper disk covered with carbon nanotube bundles have shown characteristic resonance shifts during exposure with various gas molecules. The shifts were interpreted as the change of the dielectric permittivity of the system forming the sensor due to the electric properties of the adsorbed molecules. Starting from a simplified sensor model formed by one single wall nanotube, we develop a self-consistent approach to describe the variation of the linear dielectric susceptibility of the tube at the atomic scale when molecules are adsorbed at its external surface. The sensitivity of this model sensor is tested as a function of the apolar or polar nature of the admolecules, their adsorption geometry, their concentration, and the characteristics of the tube (length, diameter,...). The comparison with data on dielectric constant changes vs adsorption, coming from measurements of the resonance frequency shifts, displays striking agreement for most of the molecular species considered. PMID- 15538889 TI - Time-dependent electron localization functions for coupled nuclear-electronic motion. AB - We study the quantum dynamics in a model system consisting of two electrons and a nucleus which move between two fixed ions in one dimension. The numerically determined wave functions allow for the calculation of time-dependent electron localization functions in the case of parallel spin and of the time-dependent antiparallel spin electron localization functions for antiparallel spin. With the help of these functions, it becomes possible to illustrate how electronic localization is modified through the vibrational wave-packet motion of the nucleus. PMID- 15538890 TI - Interaction of NaCl with solid water. AB - The interaction of NaCl with solid water, deposited on tungsten at 80 K, was investigated with metastable impact electron spectroscopy (MIES) and ultraviolet photoelectron spectroscopy (UPS) (He I). We have studied the ionization of Cl(3p) and the 1b(1), 3a(1), and 1b(2) bands of molecular water. The results are supplemented by first-principles density functional theory (DFT) calculations of the electronic structure of solvated Cl(-) ions. We have prepared NaCl/water interfaces at 80 K, NaCl layers on thin films of solid water, and H(2)O ad-layers on thin NaCl films; they were annealed between 80 and 300 K. At 80 K, closed layers of NaCl on H(2)O, and vice versa, are obtained; no interpenetration of the two components H(2)O and NaCl was observed. However, ionic dissociation of NaCl takes place when H(2)O and NaCl are in direct contact. Above 115 K solvation of the ionic species Cl(-) becomes significant. Our results are compatible with a transition of Cl(-) species from an interface site (Cl in direct contact with the NaCl lattice) to an energetically favored configuration, where Cl species are solvated. The DFT calculations show that Cl(-) species, surrounded by their solvation shell, are nevertheless by some extent accessed by MIES because the Cl(3p)-charge cloud extends through the solvation shell. Water desorption is noticeable around 145 K, but is not complete before 170 K, about 15 K higher than for pure solid water. Above 150 K the NaCl-induced modification of the water network gives rise to gas phase like structures in the water spectra. In particular, the 3a(1) emission turns into a well-defined peak. This suggests that under these conditions water molecules interact mainly with Cl(-) rather than among themselves. Above 170 K only Cl is detected on the surface and desorbs around 450 K. PMID- 15538891 TI - DNA-psoralen interaction: a single molecule experiment. AB - By attaching one end of a single lambda-DNA molecule to a microscope coverslip and the other end to a polystyrene microsphere trapped by an optical tweezers, we can study the entropic elasticity of the lambda-DNA by measuring force versus extension as we stretch the molecule. This powerful method permits single molecule studies. We are particularly interested in the effects of the photosensitive drug psoralen on the elasticity of the DNA molecule. We have illuminated the sample with different light sources, studying how the different wavelengths affect the psoralen-DNA linkage. To do this, we measure the persistence length of individual DNA-psoralen complexes. PMID- 15538892 TI - Particle connectedness and cluster formation in sequential depositions of particles: integral-equation theory. AB - We applied the integral-equation theory to the connectedness problem. The method originally applied to the study of continuum percolation in various equilibrium systems was modified for our sequential quenching model, a particular limit of an irreversible adsorption. The development of the theory based on the (quenched annealed) binary-mixture approximation includes the Ornstein-Zernike equation, the Percus-Yevick closure, and an additional term involving the three-body connectedness function. This function is simplified by introducing a Kirkwood like superposition approximation. We studied the three-dimensional (3D) system of randomly placed spheres and 2D systems of square-well particles, both with a narrow and with a wide well. The results from our integral-equation theory are in good accordance with simulation results within a certain range of densities. PMID- 15538893 TI - Network structures of polyhedral oligomeric silsesquioxane based nanocomposites: a Monte Carlo study. AB - The network structures of polyhedral oligomeric silsesquioxane based nanocomposites are studied by continuous-space Monte Carlo simulations. The nanoporous network contains intercubic pores and mesopores which can be clearly identified in this work. In terms of degree of cross linking and pore size distribution (PSD), effects of linker length, tether rigidity, and number of reactive tethers are examined. It is found that the extent of cross linking as well as the intercubic pore size of the network increases as linker length increases which are consistent with experimental findings. However, the mesopores appear to shift to a smaller radii regime for networks with longer linkers. Networks with rigid tethers contain lots of free linkers, thus, low cross linking density and narrow PSD are observed. On the other hand, reduction of the reactive tethers shows an insignificant effect on the degree of cross linking of the system. The fact that the intercubic pore size increases as the number of reactive tethers decreases causes the nanobuilding blocks to possess larger free volumes and distribute themselves more evenly throughout the system. As a result, it reduces the possibility of forming large mesopores. PMID- 15538894 TI - Ordered microstructures by assembly of ABC 3-miktoarm star terpolymers and linear homopolymers. AB - Ordered microstructures assembled from the mixture of the ABC 3-miktoarm star terpolymers and the linear homopolymers have been investigated by using dynamic density functional theory. The simulations reveal that completely different ordered microphase pattern is found with addition of a few percent homopolymers that is identical in component to one of the arms on the ABC 3-miktoarm star terpolymer. For example, the original density pattern of ABC 3-miktoarm star terpolymers with parameters of N(A)=N(B)=N(C)=10 and chi(AB)=0.90, chi(BC)=chi(CA)=0.45 is in a perfectly ordered knitting feature. However, with gradual addition of the linear polymer same as block C on ABC 3-miktoarm star terpolymer into the system, the density patterns evolve with the volume fraction of the linear polymer from the ordered knitting patterns into the hexagonal patterns. Furthermore, with addition of linear polymers same as block A, lamellar microstructure has finally resulted. The simulation points out a way for designing and manufacturing nanomaterials with totally different microstructures. PMID- 15538895 TI - Effect of ionization on the temperature- and pressure-induced phase transitions of poly(N-isopropylacrylamide) gels. AB - The ionization effects on the pressure-induced phase transition of weakly charged poly(N-isopropylacrylamide-co-acrylic acid) (PNIPA-AAc) gels have been investigated by small-angle neutron scattering. At low temperature, T, and pressure, P, the structure factor of PNIPA-AAc gels was well represented by a Lorentzian (L) function, which was similar to noncharged PNIPA gels. However, at high Ps, the contribution of inhomogeneities became large and a squared Lorentzian term had to be added in addition to the L term. At high Ts, on the other hand, a scattering maximum appeared, indicating microphase separation. This scattering maximum was suppressed by increasing P up to P approximately 100 MPa and then reincreased at higher Ps. The following facts were disclosed: (1) The peak position and height were very sensitive to P, which is mainly ascribed to strong pressure dependence of hydrophobic interaction, (2) ionization leads to microphase separation at elevated temperatures, (3) the re-entrant phase behavior is commonly observed in the P-T plane due to the parabolic variation of the polymer-solvent interaction with P, and (4) the pressure and temperature dependence of the structure factor was reproduced with the Rabin-Panyukov theory and was interpreted with a convexity of hydrophobic interaction with respect to pressure. PMID- 15538896 TI - Microrheology of entangled polymer solutions. AB - Microrheology of semidilute polymer solutions is investigated. In this paper we calculate a response function of a probe particle embedded in a semidilute polymer solution by analyzing the two-fluid model. We find that when the size of the probe particle is comparable to the viscoelastic length, the response from the longitudinal compression modes becomes more important than that of the transverse shear modes. As a result, depending on the circumstances, the obtained complex shear modulus cannot be well approximated by that measured in macroscopic rheology experiments. The present results are due to the dynamical asymmetry coupling and the existence of the cooperative dynamics, which are intrinsic to entangled polymer solutions. PMID- 15538897 TI - Texture formation under phase ordering and phase separation in polymer-liquid crystal mixtures. AB - Computational modeling of texture formation in coupled phase separation-phase ordering processes in polymer/liquid crystal mixtures is performed using a unified model based on the nematic tensor order parameter and gradient orientation elasticity. The computational methods are able to resolve defect nucleation, defect-defect interactions, and defect-particle interactions, as well as global and local morphological features in the concentration and order parameter spatiotemporal behavior. Biphasic structures corresponding to polymer dispersed liquid crystals (PDLCs), crystalline filled nematic (CFNs), and random filled nematics (RFNs) are captured and analyzed using liquid crystal defect physics and structure factors. Under spinodal decomposition due to concentration fluctuations, the PDLC structure emerges, and the nucleation and repulsive interaction of defects within nematic droplets leads to bipolar nematic droplets. Under spinodal decomposition due to ordering fluctuations, the CFNs structure emerges, and the stable polymer droplet crystal is pinned by a lattice of topological defects. For intermediate cases, where the mixture is unstable to both concentration and nematic order fluctuations, the RFN structure emerges, and polymer droplets and fibrils are pinned by a defect network, whose density increases with the curvature of the polymer-liquid crystal interface. The simulations provide an information of the role of topological defects on phase separation-phase ordering processes in polymer-liquid crystal mixtures. PMID- 15538898 TI - Theoretical strategy to provide atomistic models of comblike polymers: a generation algorithm combined with configurational bias Monte Carlo. AB - A computational strategy to model the amorphous phase of comblike polymers is presented. The strategy, denoted SuSi/CB (CB-configurational bias), combines the strength of an algorithm recently developed to generate reliable microstructures of dense amorphous polymers, which is based on a random search of energy minima, and configurational bias Monte Carlo method. The influence of different parameters used to define the characteristics of SuSi/CB on both the reliability of the generated structures and the computational effort has been examined in detail. Finally, we have modeled and characterized the supramolecular organization of poly(octadecyl acrylate) in the amorphous state. PMID- 15538899 TI - Interaction of UV radiation and inorganic carbon supply in the inhibition of photosynthesis: spectral and temporal responses of two marine picoplankters. AB - The effect of ultraviolet radiation (UVR) on inhibition of photosynthesis was studied in two species of marine picoplankton with different carbon concentration mechanisms: Nannochloropsis gaditana Lubian possesses a bicarbonate uptake system and Nannochloris atomus Butcher a CO2 active transport system. Biological weighting functions (BWFs) for inhibition of photosynthesis by UVR and photosynthesis vs irradiance (PI) curves for photosynthetically active radiation (PAR) were estimated for both species grown with an enriched CO2 supply (high dissolved inorganic carbon [DIC]: 1% CO2 in air) and in atmospheric CO2 levels (low DIC: 0.03% CO2). The response to UVR and PAR exposures was different in each species depending on the DIC treatment. Under PAR exposure, rates of maximum photosynthesis were similar between treatments in N. gaditana. However, the cultures growing in high DIC had lower sensitivity to UVR than the low DIC cultures. In contrast, N. atomus had higher rates of photosynthesis under PAR exposure with high DIC, but the BWFs were not significantly different between treatments. The results suggest that one or more processes in N. gaditana associated with HCO3- transport are target(s) for UV photodamage because there was relatively less UV inhibition of the high DIC-grown cultures in which inorganic carbon fixation is supplied by passive CO2 diffusion. Time courses of photochemical efficiency in PAR, during UV exposure and during subsequent recovery in PAR, were determined using a pulse amplitude modulated fluorometer. The results were consistent with the BWFs. In all time courses, a steady state was obtained after an initial decrease, consistent with a dynamic balance between damage and repair as found for other phytoplankton. However, the relationship of response to exposure showed a steep decline in activity that is consistent with a constant rate of repair. A novel feature of a model developed from a constant repair rate is an explicit threshold for photosynthetic response to UV. PMID- 15538900 TI - [Bye-bye to internal medicine?]. PMID- 15538901 TI - [Mortality and survival in a cohort of 1,115 HIV-infected patients (1989-97)]. AB - BACKGROUND: To study survival and HIV/AIDS-related mortality from 1989 through 1997. To analyze the effect of antiretroviral treatment and prophylaxis against P. carinii pneumonia (PCP-prophylaxis). PATIENTS AND METHODS: We retrospectively studied a cohort of 1,115 HIV (+) outpatients (331 with AIDS-defining criteria) seen in our specific HIV hospital unit from January 1989 through May 1997. We analyzed the effect of different antiretroviral treatments on annual mortality rate. In survival studies we used Cox regression analysis to analyze survival over time as well as the effect of different opportunistic events, adherence and changes in treatment during follow up. RESULTS: Mortality rate was 13.7 per 100 person-years in 1994. It went down to 4.2 during the first half of 1997 (p=0.001). Mortality rate decreased depending on treatment received: 53% (CI 95=34-65%) with monotherapy, 68% (CI 95=38-84%) with bitherapy, 86% (CI 95=40 96%) with triple therapy, and 49% (CI =29-64%) with PCP-prophylaxis. Patients with more than 100 CD4 had an increasing survival over time (p=0.002). In AIDS patients good adherence to antiretroviral treatment and PCP-prophylaxis were associated with a lower risk of death (RR=0.88; CI 95=0.63-1.22 and RR=0.72; CI 95=0.55-0.95 respectively). CONCLUSIONS: In recent years PCP-prophylaxis and antiretroviral treatment (especially combined therapy) have contributed to a decrease in AIDS-related mortality. Adherence to treatments relates to risk of death and survival. PMID- 15538902 TI - [Infectious spondylodiscitis in a health area of Galicia (Spain) from 1983-2003]. AB - OBJECTIVE: To study the clinical-epidemiological characteristics of infectious spondylodiscitis (IS) in adults of our health area in the late 20 years. MATERIAL AND METHODS: We performed a retrospective analysis of the medical records of adult patients with a diagnosis of both, tuberculous (TS) and non tuberculous spondylodiscitis (NTS), between January 1983 and December 2003. The diagnosis was made when compatible clinical-radiological picture were present in association with at least two positive blood cultures and/or micro-organism recovery from vertebral samples. Additionally, TS was diagnosed when biopsy showed typical caseating granulomas from vertebral or extra-vertebral lesions. RESULTS: 17 TS and 22 NTS were identified. The mean age was lower in patients with TS than in NTS (43.5 +/- 24.6 vs 52.0 +/- 15.2 years; mean +/- SD). Patients were predominantly males in both, TS (57%) and NTS (82%; p < 0001). The time between the onset of symptoms and diagnosis was longer in TS (16.4 +/- 15.2 weeks) than in NTS (3.9 +/- 3.2 weeks), p= 0.005. Seven (41%) of the 17 patients with TS had active extra-vertebral tuberculosis. A source of infection was presumed in 20 NTS (90%), mainly surgical spinal procedures (9/22.41%). In three TS and four NTS one o more predisposing factors were observed. The patients with NTS presented a higher prevalence of fever (41 vs 24%; p= 0.0003) and leucocytosis (41 vs 12%; p < 0.001), but less neurological impairment (9 vs 21% p= 0,01). None patient with NTS presented mixed infection and Staphylococcus aureus was the main pathogen (14/22.64%) follows by Streptococcus sp (6/22.27%). Klebsiella pneumoniae and Proteus mirabilis were the remain causative agents. Spinal cord decompression and surgical drainage of abscess were performed in five patients (24%) with TS and four patients with NTS (18%), p= 0.0027). Neurological sequels were more common in the patients with TS (24 vs 14%, p= 0.008). The global incidence of IS was 2.2 cases/105 inhabitants/year, which 1.73 cases are NTS. CONCLUSIONS: The IS are a unusual disease what occurs predominantly in male patients. In the last 10 years, a increasing prevalence of NTS was observed, with high contribution (41%) of post surgical cases. Diagnostic delay is greater in patients with TS and this condition was associated with more neurological sequels. PMID- 15538903 TI - [Generic drugs in a district hospital. Intervention strategies]. AB - OBJECTIVE: To know the prescription of generic drugs in specialized attention and the impact of two different intervention strategies: MATERIAL AND METHODS: Intervention trial with evaluation and later comparison. The data on the use of generic drugs are obtained from the medical reports. After an informative note of general character we repeated the taking of data one month later. In the service of Medicine we also analysed each doctor's profile prescriptions and a personalized informative note was edited and accompanied by a brief verbal explication of the campaign. Then repeated the collection of data to the week and the month. RESULTS: 1545 reports were revised (3460 prescribed drugs). 42% of the medication could be prescribed as generic but it was only made in 10% (21% in Internal Medicine). After the informative note no significant variations were observed. Only in Internal Medicine an initial increment was obtained (21 to 56%, RR 0.38, IC 0.26-0.55) that it disappeared at month. COMMENTS: The utility of the intervention to generic promotion is limited and transitory. Besides improve the information, seems necessary the implication of the physician in the promotion campaigns. PMID- 15538904 TI - [Health status of the oldest old: functional and medical situation in centenarians]. AB - OBJECTIVE: We planned a prospective and descriptive analysis of the centenarian population in Lugo sanitary area, establishing their social, functional and medical status as well as its relationship with their basic hematological and biochemical parameters. MATERIAL AND METHODS: During the study period between January 2001 and September 2003, participants were visited at home by a doctor and a nurse. The following variables were assessed: social status, past medical history, physical examination, functional status (Barthel index), blood analysis. RESULTS: 54 centenarians were interviewed, 16 men and 38 women. 75.9% were widows; 87% lived with their family and 57,4% in urban areas. 79.6% had followed studies. All of them had their own incomes. Regarding past medical history, 64.8% had some visual or auditive disturbances, 81.5% were taking medical drugs, 59.3% had some surgical intervention and 46.3% had been hospitalized for medical reasons. Their vaccination status was poor. Functional status, assessed by Barthel index, showed an average of 59 +/- 36.4, higher in men (82.7 +/- 28.7) than in women (49.6 +/- 35.1) (p<0.003). Blood samples were analyzed in 51 cases, there were not significant differences among them regarding sex or physical disability. CONCLUSION: The features of the centenarian population of Lugo are similar to other countries. It is an heterogeneous group. There are more women, but their clinical and functional status are significantly worse than in males. We have not founded a relationship between hematological and nutritional parameters and the degree of functional dependence in centenarians. PMID- 15538905 TI - [Cranial hyperostosis as a metastasic adenocarcinoma presentation form]. AB - Hyperostosis is a volume-unit osseous increase of very diverse etiology. We present the case of a 68-year woman with a cranial hyperostosis debuting with frontal protrusion, headache and neurologic symptoms. Image proves demonstrated a hyperostosis in the calotte and meningeal enhancement, without intracerebral lesions nor malignant cells in the cerebrospinal fluid. Analytic data were unspecific. Cranial biopsy showed huge neoplastic infiltration in bone and meninges. Primary site remained unknown after a CAT and a mammography. PMID- 15538906 TI - [Anticoagulation with bemiparina after intracerebral hemorrhage as complication of bacterial endocarditis on metallic prosthetic valve]. AB - We present the case of a patient with an intracerebral hemorrhage as complication of an infectious endocarditis (EI) produced by Neisseria sicca on a prosthetic mitral valve. The patient was anticoagulated previously with Acenocumarol as prophylaxis of thrombosis of the prosthetic valve. He was diagnosed as having IE and later he presented neurological symptoms as consequence of several intracerebral hemorrhagic foci. We decided to continue the anticoagulation with sodium heparin followed of Bemiparina and no new hemorrhagic complications nor thrombosis of the valve were observed after three months of the event. We have not found in the literature any case where low molecular weight heparin has been used as method of anticoagulation in these cases beyond two weeks. PMID- 15538907 TI - [Nocardia farcinica pneumoniae in chronic obstructive pulmonary disease patient]. AB - Nocardia farcinica is an infrequent infection that usually appears in patients with predisposing conditions, especially in immunosuppressed patients, although it has also been found in healthy individuals. Its importance as a new pathogen has been recognized only in recent years. Mainly, it affects the lung, indistinguishable from other types of pneumonia in the clinical and radiological characteristics. The main reason for detection is therapeutic failure. PMID- 15538908 TI - [The role of malnutrition and other medical factors in the evolution of patients with hip fracture]. AB - As the population progressively ages, hip fractures have become increasingly common and are associated with high morbidity and mortality and a pronounced decline in functional status. Hip fractures frequently occur in elderly patients with a high rate of comorbidity and polymedication. Patients hospitalised with hip fractures often display signs of protein malnutrition and may develop medical complications requiring intrahospital care. These factors, more than simply surgical ones, unfavourably influence the vital status and functional outcome of these patients. For this reason, it is necessary to improve the management of pre existing conditions during hospitalisation, assess and treat malnutrition and prevent medical complications to achieve optimal outcomes for these patients. With this objective, we believe that care should be provided by multidisciplinary teams in close partnership with internists. PMID- 15538909 TI - [Eosinophilic enteritis like cause of acute abdomen]. PMID- 15538910 TI - [Pneumonia as a presentation of an endobronchial chondroma]. PMID- 15538911 TI - [Pituitary apoplexy: is a surgical urgency?]. PMID- 15538912 TI - [Nontropical pyomyositis]. PMID- 15538913 TI - [Bilateral avascular necrosis at the head of the femur and human immunodeficiency virus infection]. PMID- 15538914 TI - [Left current paralysis and aortic aneurysm]. PMID- 15538915 TI - [Fever of unknown origin as presentation of chronic aortic dissection]. PMID- 15538916 TI - [Assessment of compliance with two community-acquired pneumonia-treatment protocols, implemented by the Internal Medicine Service and the Pneumology Unit]. PMID- 15538917 TI - [Spontaneous hemarthrosis associated to anticoagulant therapy]. PMID- 15538918 TI - [Lymphocytic colitis induced by carbamazepine]. PMID- 15538929 TI - The glucagonoma syndrome and necrolytic migratory erythema: a clinical review. AB - The glucagonoma syndrome is a rare disease in which a typical skin disorder, necrolytic migratory erythema, is often one of the first presenting symptoms. Weight loss and diabetes mellitus are two other prevalent characteristics of this syndrome. Necrolytic migratory erythema belongs to the recently recognized family of deficiency dermatoses of which zinc deficiency, necrolytic acral erythema and pellagra are also members. It is typically characterized on skin biopsies by necrolysis of the upper epidermis with vacuolated keratinocytes. In persistent hyperglucagonemia, excessive stimulation of basic metabolic pathways results in diabetes mellitus at the expense of tissue glycogen stores, and muscle and fat mass. Multiple (essential) nutrient and vitamin B deficiencies develop, which contribute to the dermatosis. In addition, glucagonomas may produce various other products, like pancreatic polypeptide, that add to the catabolic effects of glucagon. PMID- 15538930 TI - Follow-up and management of differentiated thyroid carcinoma: a European perspective in clinical practice. AB - As differentiated (follicular and papillary) thyroid cancer (DTC) may recur years after initial treatment, follow-up of patients with DTC is long term. However, this population has changed, with more individuals being discovered at an earlier stage of disease, so that previous follow-up protocols based mostly on data from high-risk patients no longer apply. We have proposed, in a previous issue of this Journal, an improved protocol for the follow-up of low-risk patients with DTC based on the findings of recent studies. We report here the case of a paradigmatic patient with papillary thyroid carcinoma, with the goal of illustrating the benefits of applying this algorithm in routine clinical practice. We also offer expanded and additional comments on various issues in the management of DTC. PMID- 15538931 TI - Impaired health-related quality of life in Graves' disease. A prospective study. AB - OBJECTIVE: In the acute, thyrotoxic phase, patients with Graves' disease often have both thyrotoxic and neuropsychiatric symptoms. The purpose of this prospective study was to examine health-related quality of life (HRQOL) in newly diagnosed and untreated Graves' patients and the effect of antithyroid medical treatment on HRQOL. In addition, we examined the potential influence of thyroid hormones and psychiatric symptoms on the impairment of HRQOL in the thyrotoxic phase. METHODS: A total of 30 consecutively referred patients with newly diagnosed and untreated Graves' disease and 34 age-, sex- and education-matched healthy volunteers were included in the study. HRQOL was assessed with the Medical Outcome Study 36-item Short-Form Health Status Survey (SF-36) before treatment, after reaching euthyroidism and 1 year after initiation of treatment. RESULTS: In the thyrotoxic phase of Graves' disease, HRQOL was significantly impaired, in physical, mental and social dimensions. After reaching euthyroidism, the patients reported much fewer limitations on the subscales of SF-36. One year after initiation of treatment, all SF-36 scores had normalized. However, in some patients, HRQOL continues to be impaired even 1 year after initiation of treatment, as reviewed by the individual analysis. The reduced HRQOL in the acute phase of Graves' disease was correlated to depressive and anxiety symptoms, but thyroid-associated orbitopathy also influenced HLQOL. CONCLUSIONS: Impaired HRQOL is common in the acute phase of Graves' disease. A significant proportion of the patients demonstrated persistent HRQOL impairment 1 year after initiation of treatment. Improvement of HRQOL in these patients remains a challenge for the clinician. PMID- 15538932 TI - Low triiodothyronine (T3) state: a predictor of outcome in respiratory failure? Results of a clinical pilot study. AB - BACKGROUND AND AIM: Various low triiodothyronine (T3) states have been described in severe nonthyroidal diseases and associated with a poor prognosis in cardiovascular disease patients. We assessed thyroid function in patients with severe respiratory failure from pulmonary disorders, and needing invasive or noninvasive mechanical ventilation, in order to evaluate the prognostic value of nonthyroidal illness syndrome. METHODS: We studied 32 consecutive patients with acute or acute-on-chronic respiratory failure. Measured variables upon admission included APACHE II score, the ratio of the partial pressure of oxygen in arterial blood to the fraction of oxygen in inspired gas (PaO(2)/FiO(2)), and plasma levels of free T3 (fT3) and free thyroxine (fT4), and TSH levels. Thyroid function was further evaluated at discharge. RESULTS: Plasma levels of fT3 were below normal in 17 patients (53%). Plasma fT3 was correlated with PaO(2)/FiO(2) (P<0.001), and with APACHE II score (P=0.003). In four patients (12.5%) who died, fT3 levels were significantly lower (P=0.002) than in patients who survived. In univariate logistic regression analysis, fT3 was the only factor significantly associated with an increased risk of death (odds ratio, 64.23; 95% confidence interval, 1.78-2316.86, P=0.023). Normalization of thyroid function was observed at discharge with a significant correlation between the percent increase in both fT3 and PaO(2)/FiO(2) (P=0.015). P values were calculated using Spearman's Correlation Coefficient. CONCLUSION: Our preliminary data suggest that the low T3 state is a predictor of outcome in pulmonary patients with respiratory failure. PMID- 15538933 TI - Blockade of endogenous growth hormone-releasing hormone receptors dissociates nocturnal growth hormone secretion and slow-wave sleep. AB - OBJECTIVES: A temporal association between non-rapid eye movement (NREM) sleep stages 3 and 4 and nocturnal augmentation of GH release was found long ago, yet the precise mechanism for this association has not been identified. It has been shown, however that pulsatile GHRH administration increases both slow-wave sleep (SWS) and GH. Based on these data, a role for GHRH as an inducer of SWS was proposed. To test this hypothesis, we have performed the corollary experiment whereby the action of endogenous GHRH has been antagonized. DESIGN: Healthy men (20-33 years old) had an infusion of GHRH antagonist ((N-Ac-Tyr(1), D-Arg(2)) GHRH-29 (NH(2))) or saline for a 12-h period, between 2100 and 0900 h. An i.v. bolus of GHRH was given at 0700 h and GH samples were drawn from 0700 to 0900 h to document the efficacy of GH suppression by the GHRH antagonist. METHODS: A limited montage sleep study was recorded from 2300 to 0700 h during each admission. Plasma GH concentrations were analyzed by the use of a sensitive chemiluminometric assay. RESULTS: Effectiveness of the GHRH antagonist was validated in all subjects by demonstrating 93+/-1.8% (P=0.012) suppression of GH response to a GHRH bolus. Polysomnography demonstrated that the percentage of SWS was not different when saline and GHRH antagonist nights were compared (P=0.607); other quantifiable sleep parameters were also unchanged. CONCLUSIONS: We conclude that endogenous GHRH is indispensable for the nocturnal augmentation of GH secretion, but that it is unlikely to participate in the genesis of SWS. PMID- 15538934 TI - Early treatment with GH alone in Turner syndrome: prepubertal catch-up growth and waning effect. AB - OBJECTIVE: In order to ascertain the advantages of early GH treatment in Turner syndrome (TS), we started a prospective study aimed at evaluating prepubertal height gain in a cohort of 29 girls who were treated with the same pro-kilo GH dose (1.0 IU/kg per week) since they were less than 6 years old and for at least 5 years before entering puberty. PATIENTS AND DESIGN: Following a minimum of 6 months of baseline observations, 29 girls with TS were enrolled for this prospective study provided that they (a) were less than 6 years old, (b) were below -1.0 standard deviation score (SDS) for height, (c) had a projected adult height (PAH) lower than the respective target height (TH) and (d) had a height velocity (HV) lower than -1.0 SDS. All the selected girls underwent a 5-year treatment with biosynthetic GH at a stable dose of 1.0 IU/kg per week and were periodically measured during the treatment period in order to evaluate height, HV and PAH. RESULTS: After a dramatic acceleration during the 1st year, HV was attenuated during the subsequent years, reaching its nadir at the 5th year. Height deficiency under therapy progressively decreased from entry onwards, shifting from -2.4+/-0.7 to -1.0+/-1.2 SDS. In the same period, mean PAH progressively increased, although after 5 years it remained lower than the average TH. CONCLUSIONS: (a) An effective growth-promoting strategy in TS should be based on early GH treatment, as suggested by our results. (b) This strategy could result in a prepubertal normalization of height, thus allowing the appropriate timing for the induction of puberty. (c) An initial GH dose of 1.0 IU/kg per week may be suitable during the first years of therapy, as shown by our data documenting an important waning effect of GH therapy only after the 4th year of treatment. (d) No acceleration of bone maturation was observed under this treatment regimen. PMID- 15538935 TI - Circulating levels of active ghrelin is associated with abdominal adiposity, hyperinsulinemia and insulin resistance in patients with type 2 diabetes mellitus. AB - OBJECTIVE: To investigate the relationship between the circulating level of active ghrelin and abdominal adiposity, serum levels of insulin or insulin resistance in patients with type 2 diabetes mellitus. DESIGN: We measured the plasma levels of the active form of ghrelin in 18 obese and 18 nonobese patients with type 2 diabetes mellitus using a radioimmunoassay (RIA) kit. Body fat accumulation was measured by computed tomography (CT) and insulin resistance by the glucose infusion rate (GIR) during an euglycemic hyperinsulinemic clamp study. RESULTS: Plasma levels of ghrelin in obese patients with type 2 diabetes mellitus were significantly decreased compared with nonobese patients. There were significant correlations between the plasma levels of ghrelin and BMI (r=-0.505, P<0.01), visceral (r=-0.444, P<0.01), subcutaneous (r=-0.506, P<0.01) and total (r=-0.534, P<0.01) fat area, serum levels of insulin (r=-0.513, P<0.01) or GIR (r=0.478, P<0.01) in type 2 diabetic patients. The plasma level of ghrelin was significantly associated with serum levels of insulin (F=8.468, P<0.05) or GIR (F=8.522, P<0.05) after adjustment for BMI in patients with type 2 diabetes mellitus. CONCLUSIONS: Decreased plasma levels of active ghrelin are significantly associated with abdominal adiposity, hyperinsulinemia and insulin resistance in type 2 diabetic patients. Hyperinsulinemia associated with insulin resistance may suppress plasma levels of active ghrelin in patients with type 2 diabetes mellitus. PMID- 15538936 TI - Greater use of insulin by southern European compared with Anglo-Celt patients with type 2 diabetes: the Fremantle Diabetes Study. AB - OBJECTIVE: To investigate the relationship between blood glucose-lowering therapy, glycaemia and ethnicity in urban Australians with type 2 diabetes. DESIGN: Prospective observational community-based study of diabetes care, control and complications. METHODS: We analysed cross-sectional data from 1057 patients, 238 from a southern European (SE) migrant background and 819 Anglo-Celts (AC). Follow-up data were available for 539 patients (113 SE, 426 AC) who had annual reviews over 4 years. RESULTS: The SE patients were of similar age to the AC patients but had longer diabetes duration, were less fluent in English and had less formal education. After adjustment for diabetes duration, glycosylated haemoglobin and glutamic acid decarboxylase antibody positivity in a logistic regression model, insulin use at study entry was approximately twice as frequent amongst SE as AC patients (odds ratio (95% confidence interval); 1.90 (1.20 3.02)). In the prospective arm, progression to insulin increased in both groups, from 18.0% at baseline to 22.1% at 4 years in SE and from 7.1% to 14.4% in AC patients. beta-cell function (%B) and insulin sensitivity (%S) using the homoeostasis model assessment in a subset of diet-treated patients at baseline showed that SE ethnicity was associated with lower %B and greater %S than in the AC group. CONCLUSIONS: SE patients with early non-antibody-mediated beta-cell failure progress to insulin requirement within the first 4-5 years of type 2 diabetes. This could reflect either a longer period of undiagnosed diabetes or a more rapid loss of beta-cell function after diagnosis. PMID- 15538937 TI - Thyrotropin-producing pituitary adenoma associated with Graves' disease. AB - OBJECTIVES: The examination of potential associations between Graves' disease and thyrotropin-producing pituitary adenoma (TSHoma) after treatment using octreotide, and of the expression of peroxisome proliferator-activated receptor gamma (PPAR gamma). DESIGN AND METHODS: A specimen of resected TSHoma tissue from our case was immunohistochemically examined for expression of somatostatin receptor 2A (SSTR2A) and PPAR gamma. Specimens of thyroid tissue from two cases with Hashimoto's thyroiditis were immunohistochemically examined for expression of SSTR2A. RESULTS: Expression of SSTR2A and PPAR gamma was identified in TSHoma cells. SSTR2A was also expressed in lymphocytes that had infiltrated thyroid tissue in Hashimoto's thyroiditis. In previous reports, three of four patients with TSHoma displayed Graves' disease after tumor resection, and TSH is also known to play a major role in regulating immunomodulatory gene expression in thyrocytes. CONCLUSIONS: Both the immunomodulatory effects of octreotide on intrathyroidal lymphocytes and rapid reductions in TSH may contribute to the onset of Graves' disease. Patients with TSHoma-associated autoimmune thyroiditis should undergo careful follow-up for development of Graves' disease after treatment. Both octreotide and the PPAR gamma receptor-activating ligands, thiazolidinediones, may be effective for patients with TSHoma. PMID- 15538938 TI - Involvement of the gp130 cytokine transducer in MtT/S pituitary somatotroph tumour development in an autocrine-paracrine model. AB - OBJECTIVE: gp130 cytokines are placed as auto-paracrine regulators of pituitary function, since they, as well as their receptors, have been shown to be expressed in and to act in normal and tumoral anterior pituitary cells. The objective of this work was to study their involvement in a model that shows the interaction between different cellular types that participate in a tumorigenic process. DESIGN: The dependence of a pituitary somatotrophic cell line (MtT/S) on a gp130 cytokine-producing folliculostellate (FS) cell line (TtT/GF) for tumorigenesis in vivo has been described. In order to study the participation of gp130 cytokines in the auto-paracrine stimulation of MtT/S growth, we generated MtT/S gp130 sense (gp130-S) and gp130 antisense (gp130-AS) clones stably transfected with pcDNA3/gp130 sense and pcDNA3/gp130 antisense vectors respectively. METHODS AND RESULTS: Functional characterization studies revealed that gp130-AS clones have an inhibited gp130 signalling, and proliferation studies showed that they have an impaired response to gp130 cytokines but respond normally to other independent stimuli. When injected into nude mice, MtT/S clones respond differently depending on cell number; at high concentrations MtT/S clones alone generated tumours equivalent in size to tumours derived from MtT/S plus TtT/GF cells. At low concentrations, MtT/S sense and control clones generated tumours of smaller size than tumours derived from these same clones plus TtT/GF cells, showing a dependence on FS cells. In both cases MtT/S gp130-AS clones had impaired tumour development. Furthermore, vessel density was significantly lower in tumours derived from gp130-AS plus TtT/GF cells. CONCLUSIONS: This study underlines the importance of gp130 cytokines in proliferation and establishes its role in auto paracrine pituitary growth regulation. PMID- 15538939 TI - Identification of a novel mutation in the arginine vasopressin-neurophysin II gene affecting the sixth intrachain disulfide bridge of the neurophysin II moiety. AB - OBJECTIVE: Most mutations of the arginine vasopressin-neurophysin II (AVP-NPII) gene cause autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI). Such mutations are predicted to alter the three-dimensional structure of the prohormone, which accumulates in the cell body, ultimately leading to neuronal degeneration and hormonal deficit. In this study we describe the case of a 26-year-old female reporting a long-lasting history of polyuria/polydipsia. The father of the patient was affected by diabetes insipidus and was under desmopressin treatment until the time of his death. Nevertheless, the patient had never been subjected to endocrine evaluation. DESIGN AND METHODS: Clinical and genetic studies were performed. An 8-h fluid deprivation test plus desmopressin challenge and a 5% saline solution test were performed, in order to confirm the diagnosis. DNA was extracted from peripheral blood lymphocytes and subjected to direct sequencing of the entire coding region of the AVP-NPII gene. RESULTS AND CONCLUSIONS: Clinical assessment of the patient confirmed the diagnosis of neurohypophyseal diabetes insipidus. Desmopressin treatment was started, which effectively reversed the polyuria/ polydipsia syndrome. Genetic analysis revealed a novel mutation (1665T>A) in exon 2 of the AVP-NPII gene, disrupting one of the disulfide bonds present in the NPII moiety which play a fundamental role in determining the proper folding of the molecule. In summary, in the present study we have described a novel mutation of the AVP-NPII gene, which is consistent with the malfolding/toxicity hypothesis underlying the pathogenesis of adFNDI. PMID- 15538940 TI - Ectopic Agouti protein overexpression increases stimulated corticosterone production without effect on adenylate cyclase activity in mouse adrenal cells. AB - OBJECTIVE: The antagonism of Agouti protein (AP) and Agouti-related protein on melanocortin receptors suggests an inhibitory role in the regulation of steroidogenesis. However, we have previously demonstrated that ectopic AP overexpression increased restraint-induced corticosterone release and adrenal reactivity to ACTH in mice. A high steroidogenic response to ACTH may be a consequence of a stimulatory AP action on the adenylate cyclase (AC) and/or intracellular steroidogenic enzymes. The aim of the present study was to estimate the effect of ectopic AP overexpression on the activity of AC and steroidogenic intracellular enzymes. METHODS: ACTH and forskolin were used for AC stimulation, and dibutyryl cAMP and progesterone were used for stimulation of intracellular steroidogenic enzymes in isolated adrenal cells in male C57Bl/6J mice of two Agouti genotypes: A(y)/a (ectopic AP overexpression) and a/a (absence of AP in all tissues). RESULTS: ACTH and forskolin increased cAMP accumulation to the same extent in both A(y)/a and a/a mouse adrenal cells (P<0.001; ANOVA), but resulted in higher corticosterone production in A(y)/a mice (P<0.001 for ACTH and P<0.01 for forskolin; ANOVA). Dibutyryl cAMP- and progesterone-induced corticosterone production was higher in A(y)/a mice than in a/a mice (P<0.001 for dibutyryl cAMP and P<0.01 for progesterone; ANOVA). CONCLUSIONS: Ectopic AP overexpression increased stimulated corticosterone production and intracellular steroidogenic enzyme reactivity to cAMP without an effect on AC activity. PMID- 15538941 TI - Gonadotropin-releasing hormone (GnRH) agonist triptorelin inhibits estradiol induced serum response element (SRE) activation and c-fos expression in human endometrial, ovarian and breast cancer cells. AB - BACKGROUND AND METHODS: The majority of human endometrial (>80%), ovarian (>80%) and breast (>50%) cancers express GnRH receptors. Their spontaneous and epidermal growth-factor-induced proliferation is dose- and time-dependently reduced by treatment with GnRH and its agonists. In this study, we demonstrate that the GnRH agonist triptorelin inhibits estradiol (E2)-induced cancer cell proliferation. RESULTS: The proliferation of quiescent estrogen receptor alpha (ER alpha)-/ER beta-positive, but not of ER alpha-negative/ER beta-positive endometrial, ovarian and breast cancer cell lines, was significantly stimulated (P<0.001) (ANOVA) after treatment with E2 (10(-8) M). This effect was time- and dose-dependently antagonized by simultaneous treatment with triptorelin. The inhibitory effect was maximal at 10(-5) M concentration of triptorelin (P<0.001). In addition, we could show that, in ER alpha-/ER beta-positive cell lines, E2 induces activation of serum response element (SRE) and expression of the immediate early-response gene c-fos. These effects were blocked by triptorelin (P<0.001). E2-induced activation of estrogen-response element (ERE) was not affected by triptorelin. CONCLUSIONS: The transcriptional activation of SRE by E2 is due to ER alpha activation of the mitogen-activated protein kinase (MAPK) pathway. This pathway is impeded by GnRH, resulting in a reduction of E2-induced SRE activation and, in consequence, a reduction of E2-induced c-fos expression. This causes downregulation of E2 induced cancer cell proliferation. PMID- 15538942 TI - Tumor necrosis factor-alpha (TNF-alpha) promotes cell survival during spermatogenesis, and this effect can be blocked by infliximab, a TNF-alpha antagonist. AB - OBJECTIVE: Tumor necrosis factor-alpha (TNF-alpha) has been shown to inhibit germ cell death in human seminiferous epithelium. In the present study, we wanted to explore the effects of TNF-alpha in the rat seminiferous epithelium and to study molecular mechanisms of germ cell apoptosis. Furthermore, the effects of infliximab were studied. Infliximab is a TNF-alpha antagonist used in autoimmune disorders, such as rheumatoid arthritis and Crohn's disease. METHODS: Rat seminiferous tubule segments were cultured in the presence and absence of TNF alpha, infliximab and SN50, a NF-kappa B inhibitor. TUNEL-staining and cleaved caspase-3 immunohistochemistry combined with squash preparations of rat seminiferous tubule segments were used to evaluate the number of apoptotic cells. Western blot analyses were performed on cultured seminiferous tubule segments for Bcl-2 family proteins (Bax, Bad, Bcl-w, Bcl-xL) and fas ligand. RESULTS: TNF alpha promotes cell survival in the rat seminiferous epithelium, and this prosurvival effect can be blocked by infliximab, a TNF-alpha antagonist. Bcl-xL was found to be upregulated in mitochondrial membranes by TNF-alpha, and this upregulation was inhibited by infliximab. Inhibition of NF-kappa B translocation to the nucleus prevented the prosurvival effect of TNF-alpha on seminiferous epithelium. CONCLUSIONS: The present study demonstrates that TNF-alpha promotes cell survival in the rat seminiferous epithelium, and this effect can be blocked by infliximab. This is the first study to show the effects of infliximab in the testis. The prosurvival effect of TNF-alpha might be at least partly mediated by modulating the expression and subcellular localization of Bcl-2 family proteins. PMID- 15538943 TI - Vitamin D receptor polymorphisms: are they really associated with type 1 diabetes? PMID- 15538944 TI - Volume expansion and plasma protein clearance during intravenous infusion of 5% albumin and autologous plasma. AB - Autologous plasma may be used to replace plasma volume and plasma proteins during surgery, but its effectiveness is largely unknown. In the present study, the characteristics of predonated frozen and thawed autologous plasma were compared with those of 5% albumin in 15 male volunteers who received 10 ml/kg of body weight of these colloids as intravenous infusions over 30 min. Venous blood was sampled and urine was collected over 8 h to outline the volume expansion and blood-interstitial fluid space transport of three plasma proteins (albumin, fibrinogen and antithrombin) by means of mass balance analysis. The maximum plasma dilution of 5% albumin and autologous plasma averaged 17 and 21% respectively, and their half-lives were 2.5 and 2.9 h respectively (P<0.03). The between-subject variability in dilution was most pronounced for autologous plasma. Transport of protein from blood to the interstitial space occurred faster when the infused fluid contained the protein in question. The rate was highest at 60 min, and the process was still in progress at 8 h when approx. 60% of the infused albumin, 45% of the fibrinogen and 75% of the infused antithrombin had been translocated to the interstitial fluid space. In contrast with the proteins, excess plasma water was removed by urinary excretion. It is concluded that the volume expansion is equivalent for the two colloid fluids, although it is more predictable for 5% albumin. The transport of protein outlasted the volume expansion. PMID- 15538946 TI - Early and long-term outcome of elective stenting of the infarct-related artery in patients with viability in the infarct-area: Rationale and design of the Viability-guided Angioplasty after acute Myocardial Infarction-trial (The VIAMI trial). AB - BACKGROUND: Although percutaneous coronary intervention (PCI) is becoming the standard therapy in ST-segment elevation myocardial infarction (STEMI), to date most patients, even in developed countries, are reperfused with intravenous thrombolysis or do not receive a reperfusion therapy at all. In the post-lysis period these patients are at high risk for recurrent ischemic events. Early identification of these patients is mandatory as this subgroup could possibly benefit from an angioplasty of the infarct-related artery.Since viability seems to be related to ischemic adverse events, we initiated a clinical trial to investigate the benefits of PCI with stenting of the infarct-related artery in patients with viability detected early after acute myocardial infarction. METHODS: The VIAMI-study is designed as a prospective, multicenter, randomized, controlled clinical trial. Patients who are hospitalized with an acute myocardial infarction and who did not have primary or rescue PCI, undergo viability testing by low-dose dobutamine echocardiography (LDDE) within 3 days of admission. Consequently, patients with demonstrated viability are randomized to an invasive or conservative strategy. In the invasive strategy patients undergo coronary angiography with the intention to perform PCI with stenting of the infarct related coronary artery and concomitant use of abciximab. In the conservative group an ischemia-guided approach is adopted (standard optimal care).The primary end point is the composite of death from any cause, reinfarction and unstable angina during a follow-up period of three years. CONCLUSION: The primary objective of the VIAMI-trial is to demonstrate that angioplasty of the infarct related coronary artery with stenting and concomitant use of abciximab results in a clinically important risk reduction of future cardiac events in patients with viability in the infarct-area, detected early after myocardial infarction. PMID- 15538945 TI - Stimulation of allergen-loaded macrophages by TLR9-ligand potentiates IL-10 mediated suppression of allergic airway inflammation in mice. AB - BACKGROUND: Previously, we demonstrated that OVA-loaded macrophages (OVA-Mphi) partially suppress OVA-induced airway manifestations of asthma in BALB/c mice. In vitro studies showed that OVA-Mphi start to produce IL-10 upon interaction with allergen-specific T cells, which might mediate their immunosuppressive effects. Herein, we examined whether IL-10 is essential for the immunosuppressive effects of OVA-Mphi in vivo, and whether ex vivo stimulation of the IL-10 production by OVA-Mphi could enhance these effects. METHODS: Peritoneal Mphi were loaded with OVA and stimulated with LPS or immunostimulatory sequence oligodeoxynucleotide (ISS-ODN) in vitro. The increase of IL-10 production was examined and, subsequently, ex vivo stimulated OVA-Mphi were used to treat (i.v.) OVA sensitized mice. To further explore whether Mphi-derived IL-10 mediates the immunosuppressive effects, Mphi isolated from IL-10-/- mice were used for treatment. RESULTS: We found that stimulation with LPS or ISS-ODN highly increased the IL-10 production by OVA-Mphi (2.5-fold and 4.5-fold increase, respectively). ISS-ODN stimulation of OVA-Mphi significantly potentiated the suppressive effects on allergic airway inflammation. Compared to sham-treatment, ISS-ODN-stimulated OVA-Mphi suppressed the airway eosinophilia by 85% (vs. 30% by unstimulated OVA-Mphi), IL-5 levels in bronchoalveolar lavage fluid by 80% (vs. 50%) and serum OVA-specific IgE levels by 60% (vs. 30%). Importantly, IL-10-/ Mphi that were loaded with OVA and stimulated with ISS-ODN ex vivo, failed to suppress OVA-induced airway inflammation. CONCLUSIONS: These results demonstrate that Mphi-derived IL-10 mediates anti-inflammatory responses in a mouse model of allergic asthma, which both can be potentiated by stimulation with ISS-ODN. PMID- 15538947 TI - No effects of GSM-modulated 900 MHz electromagnetic fields on survival rate and spontaneous development of lymphoma in female AKR/J mice. AB - BACKGROUND: Several reports indicated that non-thermal electromagnetic radiation such as from mobile phones and base stations may promote cancer. Therefore, it was investigated experimentally, whether 900 MHz electromagnetic field exposure influences lymphoma development in a mouse strain that is genetically predisposed to this disease. The AKR/J mice genome carries the AK-virus, which leads within one year to spontaneous development of thymic lymphoblastic lymphoma. METHODS: 320 unrestrained female mice were sham-exposed or exposed (each n = 160 animals) to GSM like 900 MHz electromagnetic fields for 24 hours per day, 7 days per week, at an average whole body specific absorption rate (SAR) value of 0.4 W/kg. Animals were visually checked daily and were weighed and palpated weekly. Starting with an age of 6 months, blood samples were taken monthly from the tail. Animals with signs of disease or with an age of about 46 weeks were sacrificed and a gross necropsy was performed. RESULTS: Electromagnetic field exposure had a significant effect on body weight gain, with higher values in exposed than in sham-exposed animals. However, survival rate and lymphoma incidence did not differ between exposed and sham-exposed mice. CONCLUSION: These data do not support the hypothesis that exposure to 900 MHz electromagnetic fields is a significant risk factor for developing lymphoma in a genetically predisposed species, even at a relatively high exposure level. PMID- 15538949 TI - Thromboembolic events and haematological diseases: a case of stroke as clinical onset of a paroxysmal nocturnal haemoglobinuria. AB - Some haematological diseases are associated to an increased risk of thromboembolic events. We report a case of paroxysmal nocturnal haemoglobinuria (PNH) in which a cerebrovascular event represented the first clinical manifestation of disease. PNH is associated to thromboembolic events, generally of venous districts often involving unusual locations such as mesenteric vessels, sagittal veins, inferior vena cava and renal veins.To our knowledge arterial thrombotic episodes are rare and the involvement of arterial cerebral vessels is exceptional. Then, our case points out the importance of investigating about haematological disorders in all patients presenting with a stroke, in which the common predisposing conditions are excluded. PMID- 15538948 TI - Bi-directional modulation of AMPA receptor unitary conductance by synaptic activity. AB - BACKGROUND: Knowledge of how synapses alter their efficiency of communication is central to the understanding of learning and memory. The most extensively studied forms of synaptic plasticity are long-term potentiation (LTP) and its counterpart long-term depression (LTD) of AMPA receptor-mediated synaptic transmission. In the CA1 region of the hippocampus, it has been shown that LTP often involves a rapid increase in the unitary conductance of AMPA receptor channels. However, LTP can also occur in the absence of any alteration in AMPA receptor unitary conductance. In the present study we have used whole-cell dendritic recording, failures analysis and non-stationary fluctuation analysis to investigate the mechanism of depotentiation of LTP. RESULTS: We find that when LTP involves an increase in unitary conductance, subsequent depotentiation invariably involves the return of unitary conductance to pre-LTP values. In contrast, when LTP does not involve a change in unitary conductance then depotentiation also occurs in the absence of any change in unitary conductance, indicating a reduction in the number of activated receptors as the most likely mechanism. CONCLUSIONS: These data show that unitary conductance can be bi-directionally modified by synaptic activity. Furthermore, there are at least two distinct mechanisms to restore synaptic strength from a potentiated state, which depend upon the mechanism of the previous potentiation. PMID- 15538950 TI - Assessment of possible impact of a health promotion program in Korea from health risk trends in a longitudinally observed cohort. AB - BACKGROUND: Longitudinally observed cohort data can be utilized to assess the potential for health promotion and healthcare planning by comparing the estimated risk factor trends of non-intervened with that of intervened. The paper seeks (1) to estimate a natural transition (patterns of movement between states) of health risk state from a Korean cohort data using a Markov model, (2) to derive an effective and necessary health promotion strategy for the population, and (3) to project a possible impact of an intervention program on health status. METHODS: The observed transition of health risk states in a Korean employee cohort was utilized to estimate the natural flow of aggregated health risk states from eight health risk measures using Markov chain models. In addition, a reinforced transition was simulated, given that a health promotion program was implemented for the cohort, to project a possible impact on improvement of health status. An intervened risk transition was obtained based on age, gender, and baseline risk state, adjusted to match with the Korean cohort, from a simulated random sample of a US employee population, where a health intervention was in place. RESULTS: The estimated natural flow (non-intervened), following Markov chain order 2, showed a decrease in low risk state by 3.1 percentage points in the Korean population while the simulated reinforced transition (intervened) projected an increase in low risk state by 7.5 percentage points. Estimated transitions of risk states demonstrated the necessity of not only the risk reduction but also low risk maintenance. CONCLUSIONS: The frame work of Markov chain efficiently estimated the trend, and captured the tendency in the natural flow. Given only a minimally intense health promotion program, potential risk reduction and low risk maintenance was projected. PMID- 15538951 TI - Does the 12-item General Health Questionnaire contain multiple factors and do we need them? AB - BACKGROUND: The 12-item General Health Questionnaire (GHQ-12) is widely used as a unidimensional instrument, but factor analyses tended to suggest that it contains two or three factors. Not much is known about the usefulness of the GHQ-12 factors, if they exist, in revealing between-patient differences in clinical states and health-related quality of life. METHODS: We addressed this issue in a cross-sectional survey of out-patients with psychological disorders in Singapore. The participants (n = 120) completed the GHQ-12, the Beck Anxiety Inventory, and the Short-Form 36 Health Survey. Confirmatory factor analysis was used to compare six previously proposed factor structures for the GHQ-12. Factor scores of the best-fitting model, as well as the overall GHQ-12 score, were assessed in relation to clinical and health-related quality of life variables. RESULTS: The 3 factor model proposed by Graetz fitted the data better than a unidimensional model, two 2-factor models, and two other 3-factor models. However, the three factors were strongly correlated. Their values varied in a similar fashion in relation to clinical and health-related quality of life variables. CONCLUSIONS: The 12-item General Health Questionnaire contains three factors, namely Anxiety and Depression, Social Dysfunction, and Loss of Confidence. Nevertheless, using them separately does not offer many practical advantages in differentiating clinical groups or identifying association with clinical or health-related quality of life variables. PMID- 15538952 TI - S-adenosylmethionine (SAM-e) for the treatment of depression in people living with HIV/AIDS. AB - BACKGROUND: This study reports on clinical data from an 8-week open-label study of 20 HIV-seropositive individuals, diagnosed with Major Depressive Disorder (DSM IV), who were treated with SAM-e (S-Adenosylmethionine). SAM-e may be a treatment alternative for the management of depression in a population reluctant to add another "pill" or another set of related side effects to an already complex highly active antiretroviral therapy (HAART) regimen. METHODS: The Hamilton Rating Scale for Depression (HAM-D) and the Beck Depression Inventory (BDI) were used to assess depressive symptomatology from 1,2,4,6 and 8 weeks after initiation of treatment with SAM-e. RESULTS: Data show a significant acute reduction in depressive symptomatology, as measured by both the HAM-D and the BDI instruments. CONCLUSIONS: SAM-e has a rapid effect evident as soon as week 1 (p < .001), with progressive decreases in depression symptom rating scores throughout the 8 week study. PMID- 15538953 TI - AIDS-defining illnesses among patients with HIV in Singapore, 1985 to 2001: results from the Singapore HIV Observational Cohort Study (SHOCS). AB - BACKGROUND: The objective was to describe the causes of initial and overall AIDS defining disease episodes among HIV patients in Singapore. METHODS: A retrospective observational cohort study was performed of all adult patients seen at the national HIV referral center between 1985 and 2001. Data were extracted from the patients' records by ten trained healthcare workers. AIDS-defining conditions were established using predefined criteria. RESULTS: Among 1504 patients, 834 had experienced one or more AIDS-defining diseases. The most frequent causes of the initial AIDS-defining episode were Pneumocystis carinii pneumonia (35.7%), Mycobacterium tuberculosis (22.7%) and herpes simplex (7.4%). In total 1742 AIDS-defining episodes occurred. The most frequent causes were Pneumocystis carinii pneumonia (25.1%), Mycobacterium tuberculosis (16.2%) and cytomegalovirus retinitis (9.5%). CONCLUSIONS: The most frequent causes of AIDS defining illnesses in Singapore are similar to those reported in the West, prior to the introduction of anti-retroviral therapy. Opportunistic infections remain the most frequent AIDS-defining illnesses. PMID- 15538954 TI - Chronic idiopathic thrombocytopenic purpura (ITP): molecular mechanisms and implications for therapy. AB - Chronic idiopathic thrombocytopenic purpura (ITP) is an immune-mediated disorder in which platelets are prematurely destroyed in the reticuloendothelial system by platelet autoantibodies. However, it is becoming clear that the pivotal process of the humoral immune response in the pathogenesis of the disorder is a complex interaction between antigen-presenting cells, T cells and B cells. Furthermore, it is increasingly evident that regulatory T cells play an important role and that T-cell-mediated cytotoxicity contributes to the destruction of platelets in ITP. Different new approaches to immunotherapy in chronic ITP have been explored, including use of anti-CD20, anti-CD154 and anti-CD52 antibodies. So far, these therapies have been antigen-nonspecific and the risk of general immunosuppression is a concern. Thus, improving our understanding of the interaction and relative contribution of humoral and cell-mediated mechanisms is essential for developing antigen-specific immunotherapies for the treatment of this disorder. This review aims to elucidate the current status of knowledge of the cellular and humoral immune components of chronic ITP, together with the implications of this knowledge for therapy. PMID- 15538955 TI - Pharmacokinetics and excretion of gamma-hydroxybutyrate (GHB) in healthy subjects. AB - In Europe and the United States, the recreational use of gamma-hydroxy butyric acid (GHB) at dance clubs and "rave" parties has increased substantially. In addition, GHB is used to assist in the commission of sexual assaults. The aim of this controlled clinical study was to acquire pharmacokinetic profiles, detection times, and excretion rates in human subjects. Eight GHB-naive volunteers were administered a single 25-mg/kg body weight oral dose of GHB, and plasma, urine, and oral fluid specimens were analyzed by using gas chromatography-mass spectrometry (GC-MS). Liquid-liquid extraction was performed after acid conversion of GHB to gamma-butyrolactone. Limits of quantitation of 0.1 (oral fluid), 0.2 (urine), and 0.5 microg/mL (plasma) could be achieved in the selected ion monitoring mode. GHB plasma peaks of 39.4 +/- 25.2 microg/mL (mean +/- SEM) occurred 20-45 min after administration. The terminal plasma elimination half life was 30.4 +/- 2.45 min, the distribution volume 52.7 +/- 15.0 L, and the total clearance 1228 +/- 233 microL/min. In oral fluid, GHB could be detected up to 360 min, with peak concentrations of 203 +/- 92.4 microg/mL in the 10-min samples. In urine, 200 +/- 71.8 and 230 +/- 86.3 microg/mL, were the highest GHB levels measured at 30 and 60 min, respectively. Only 1.2 +/- 0.2% of the dose was excreted, resulting in a detection window of 720 min. Common side-effects were confusion, sleepiness, and dizziness; euphoria and change of vital functions were not observed. GHB is extensively metabolized and rapidly eliminated in urine and oral fluid. Consequently, samples should be collected as soon as possible after ingestion. PMID- 15538956 TI - Postmortem redistribution of the enantiomers of citalopram and its metabolites: an experimental study in rats. AB - A rat model was used to study if postmortem redistribution of the S- and R enantiomers of citalopram (CIT) and its metabolites demethylcitalopram (DCIT) and didemethylcitalopram (DDCIT) occurs after three different subcutaneous dosing procedures with racemic CIT. Two groups underwent chronic administration (20 mg/kg daily) using osmotic pumps. After 10 days, 1 of these groups received an acute-on-chronic drug challenge with a single injection of 100 mg/kg. The third group received the single 100 mg/kg dose only. Heart blood and brain samples were collected antemortem and 1, 3, or 24 h postmortem for enantioselective HPLC analysis. Increased postmortem blood drug and metabolite concentrations compared with corresponding antemortem concentrations were observed in all groups (p < 0.05 to p < 0.001). At 24 h after death, the ratios between postmortem and antemortem blood concentrations were around 3-4 for CIT as well as for the metabolites. In the brain, no major differences between antemortem and postmortem drug and metabolite concentrations were observed. The enantiomeric (S/R) concentrations ratios of CIT and metabolites in blood and brain were of similar magnitude before and after death. No differences between antemortem and postmortem parent drug-to-metabolite (P/M) ratios for CIT/DCIT in blood were observed. Finally, this animal model demonstrates that the S- and R-enantiomers of CIT and its metabolites were redistributed to the same extent postmortem. PMID- 15538957 TI - Analysis of pyrolysis products of methamphetamine. AB - This study examined the pyrolysis products of d-methamphetamine (d-MA) and the pyrolysis mechanism. A sealed glass tube, in which MA-HCl was placed, was wrapped with pyrolysis-foil and heated at the Curie point of the pyrolysis-foil. The pyrolysis products of MA were detected by gas chromatography-mass spectrometry and liquid chromatography-electrospray ionization-mass spectrometry. MA-d(3)-HCl, in which all the hydrogen atoms of N-methyl group of MA were substituted with deuterium atoms, was pyrolyzed to investigate the transformation of a methyl group of MA. Amphetamine (AM) and dimethylamphetamine (DMA) were produced via demethylation and methylation reactions, respectively, at temperatures above 315 degrees C. The demethylation and methylation reactions were the major pyrolysis processes at temperatures below 358 degrees C. At temperatures above 315 degrees C, the reaction of DMA with a methyl group eliminated from the methylamino group of MA resulted in the formation of benzylethyltrimethylammonium (BEMA). This transformation reveals that demethylation and methylation reactions occur in the form of a methyl cation. The thermal degradation of BEMA, the abstraction of a proton at the beta-position and the elimination of a trimethylamine, produced allylbenzene, cis-beta-methylstyrene, and trans-beta-methylstyrene at temperatures above 315 degrees C. At temperatures above 445 degrees C, the optical isomers, or l-isomers of AM, MA, and DMA, were produced as pyrolysis products. PMID- 15538958 TI - A reference range for endogenous gamma-hydroxybutyrate in urine by gas chromatography-mass spectrometry. AB - Gamma-hydroxybutyrate (GHB) has been implicated in drug-facilitated sexual assault (DFSA). The interpretation of GHB levels in biological samples collected for evidence is complicated by the natural presence of this compound in the body, and by its extremely rapid elimination after ingestion. There is a lack of agreement regarding a suitable cut-off concentration, which can reliably separate endogenous concentrations in urine from those reflecting ingestion. We have developed a method for the analysis of low levels of GHB in urine and have used it to establish a reference range for normal females. The method uses liquid liquid extraction, silyl-derivatization, and gas chromatographic-mass spectrometric analysis. The limit of detection was 0.1 mg/L, and the method was linear from 0.1 to 5.0 mg/L. Our analysis of 50 urine samples donated by normal women indicates an upper limit of normal for urinary GHB of 1.46 mg/L or 323 microg GHB/mmol of creatinine. We propose that a 5 mg/L cut-off for urine GHB concentration, or 1000 microg GHB/mmol creatinine, will separate endogenous GHB concentrations from those reflecting GHB ingestion in antemortem samples with greater than 99% confidence, providing that a specific assay method comparable with that we describe is used. We demonstrate that urinary GHB concentrations fall with age and that this can be corrected for by measurement of the GHB/creatinine ratio. PMID- 15538959 TI - Effectiveness of multiple internal standards: deuterated analogues of methylenedioxymethamphetamine, methylenedioxyamphetamine, methamphetamine, and amphetamine. AB - With the increasing abuse of methylenedioxymethamphetamine (MDMA) thereby requiring analysis, we have undertaken a systematic evaluation on parameters associated with the analysis of MDMA and related compounds, including methylenedioxyamphetamine (MDA), methamphetamine (MA), and amphetamine (AM). Parameters studied included three solid-phase adsorbents, five derivatization reagents, and four deuterated internal standards (IS). This report examines whether differences in quantitation data derived from the use of four ISs (one for each analyte) and two ISs (one for AM and MA, one for MDA and MDMA) are statistically significant. Two types of samples were included in this study. The first type (Type I) included four replicate sets of standard solutions prepared in urine matrix. All analytes (AM, MA, MDA, and MDMA) were included in all samples, and these analytes' concentrations in each set were at five levels (100, 250, 500, 1000, and 2000 ng/mL). Four deuterated analogues (MA-d(8), AM-d(8), MDMA-d(5), and MDA-d(5)) at 500 ng/mL were also included in all solutions. The second type of samples (Type II) included 25 case urine specimens. Most of these specimens contained MA/AM and/or MDMA/MDA. The specific objective of this study is to determine whether the 4-IS approach can indeed generate better quantitative data than a less-costly 2-IS. For Type I samples, where the true concentrations of the analytes are known, two-sample t-test is adapted to examine whether the two sets of prediction errors (i.e., known concentration minus calculated concentration) resulting from the 4-IS and the 2-IS approaches are statistically different. For Type II samples, where the analytes' true concentrations are unknown, one-sample t-test was adapted to determine whether the difference of the quantitation results derived from the 4-IS and the 2-IS approaches is statistically significant. Statistical analysis of quantitation data derived from Types I and II samples indicates that differences in MDA and MDMA concentrations resulting from the use of one (MDA-d(5) or MDMA-d(5)) or two (MDA-d(5) and MDMA d(5)) are statistically nonsignificant. On the other hand, similar analysis on data derived from Type I samples indicate the use of the analytes' respective deuterated analogues as the ISs appear to generate better quantitative data for AM and MA. PMID- 15538960 TI - The determination of cocaine, benzoylecgonine, and cocaethylene in small-volume oral fluid samples by liquid chromatography-quadrupole-time-of-flight mass spectrometry. AB - A quantitative analysis was developed for the determination of cocaine, benzoylecgonine, and cocaethylene in oral fluid using liquid chromatography tandem mass spectrometry. After internal standardization and solid-phase extraction, chromatographic separation was achieved on a reversed-phase column by gradient elution. The reconstructed mass chromatograms of the collision-induced dissociation transitions of m/z 290 --> m/z 168 (benzoylecgonine), m/z 304 --> m/z 168+119 (2'-methylbenzoylecgonine), m/z 304 --> m/z 182 (cocaine), m/z 318 - > m/z 196 (cocaethylene), and m/z 318 --> m/z 182+119 (2'-methylcocaine) were used for quantitation. The developed method was adequately validated. Good linearity was obtained from 10 to 1000 microg/L. Extraction recoveries exceeded 85% for all compounds. Excellent total and within-run reproducibilities (CV% < 20%) and accuracy figures were obtained. The limit of detection (signal-to-noise ratio >/= 3) was 1 microg/L for all three compounds. As such, a method for drug abuse confirmation analysis in oral fluid, compatible with the present day saliva collecting devices, is obtained. The method was applied to real samples (n = 15) obtained from suspected drug users, of which seven proved positive. The concentrations found in the positive samples were between 10.2 and 200.6 microg/L for cocaine, < limit of quantification (LOQ) and 10.5 microg/L for cocaethylene, and < LOQ and 59.2 microg/L for benzoylecgonine. PMID- 15538961 TI - Negative ion chemical ionization-gas chromatographic-mass spectrometric determination of residues of different pyrethroid insecticides in whole blood and serum. AB - A new rapid and sensitive analytical method using negative ion chemical ionization-gas chromatography-mass spectrometry in selective ion monitoring mode has been developed for the determination of residues of different synthetic pyrethroid insecticides, allethrin, bifenthrin, cypermethrin, cyphonothrin, cyfluthrin, lambda-cyhalothrin, deltamethrin, fenvalerate, fenpropathrin, permethrin, prallethrin, and trans-fluthrin, in whole blood. The residues of pyrethroid molecules were extracted from the whole blood using a hexane and acetone (8:2, v/v) solvent mixture without separating the serum. The method was found sensitive to detect the residues of pyrethroids up to the level 0.2 pg/mL. Experiments conducted with the whole blood samples at the fortification level 1 100 pg/mL showed 91-103% recovery, whereas blood serum samples collected after the fortification of pyrethroids in whole blood showed 36-54% recovery. Recovery experiments conducted by direct fortification of pyrethroids in blood serum samples showed 96-108%. The applications of the analytical method was tested by analyzing 73 human blood samples collected from the population exposed continuously to different pyrethroid-based formulations. None of the blood samples showed residues of pyrethroids. The results were also confirmed by the detection of the appropriate amounts in a number of these samples, which had subsequently been spiked with known quantity of pyrethroids. PMID- 15538962 TI - Detecting cocaine use through sweat testing: multilevel modeling of sweat patch length-of-wear data. AB - Although urine analysis remains the standard for detection of drugs of abuse, sweat patches provide a convenient alternative that avoids some of the problems with drug testing such as violations of privacy in observed urination, possibility of disease transmission, and transport of noxious fluids. This study examined minimum length of wear necessary to detect recent or concurrent cocaine use in a convenience sample of active cocaine users and also differences in analyte concentrations with increasing longer-term wear. Twenty-seven subjects (22 active drug users and 5 comparison subjects who did not use drugs) wore short term ((1/2)h, 1 h, 1(1/2) h, and 2 h), then long-term patches (1, 3, 7, and 14 day). Short- and long-term patches were identical except for duration of wear. The predominant analyte found was cocaine, followed by benzoylecgonine, then ecgonine methylester. The minimum duration that patches must be worn to detect recent or concurrent cocaine use in this sample is more than 2 h and less than or equal to 1 day. Analyte concentrations increase significantly with increasing lengths of wear. However, increases between the one-week and two-week patches were significant for benzoylecgonine only. PMID- 15538963 TI - Determination of three beta-blockers in biofluids and solid tissues by liquid chromatography-electrospray-mass spectrometry. AB - A LC-MS method using clenbuterol as internal standard was developed and validated for three b-blockers (BB) (atenolol, metoprolol, and propranolol) in rabbit postmortem matrices: heart, lung, kidney, liver, brain, blood, vitreous humor, gastric liquid, and urine. The BB were extracted from 2.0 mL of biofluids or 200 mg of solid tissues (after grinding and homogenization) by liquid-liquid extraction using Extrelut columns. Chromatographic separation involved a Nucleosil C18 (150 mm x 1-mm i.d., 5 microm) column together with a gradient of acetonitrile in 2mM, pH 3 ammonium formate. The compounds were ionized in the ionspray source of the atmospheric pressure mass spectrometer and fragmented by in-source collisions. The fragment ions were detected in the positive selected ion monitoring mode, targeting one quantitation and two confirmation ions per compound. The extraction recovery ranged between 10 and 40%, depending on the matrices. The limits of quantitation were 50 ng/g in tissues, 50 microg/L in blood and urine, and 10 microg/L in vitreous humor. Indeed, as preliminary results in one rabbit administered 5 mg/kg of each BB showed that BBs were more concentrated in some postmortem organs, validation was performed in the relevant concentration area in these particular tissues. The technique was found to be linear between 50 ng/g and 5000 ng/g for heart and liver, between 50 microg/L and 5000 microg/L for urine extracts, between 1000 ng/g and 50 000 ng/g for lung and kidney, and between 500 microg/L and 5000 microg/L for gastric content. A quadratic equation best fitted the calibration curve in blood between 50 microg/L and 5000 microg/L, as well as in brain between 50 ng/g and 40,000 ng/g. The correlation coefficients were all higher than 0.997. Intra- and interassay precision and accuracy fulfilled the international requirements. This simple and sensitive assay was applied to the determination of three BB in the biofluids and tissues of a rabbit as part of a preliminary step of a postmortem redistribution study and is also suitable for the routine determination of BB in forensic investigations. PMID- 15538964 TI - A rapid and sensitive ESI-MS screening procedure for ketamine and norketamine in urine samples. AB - Traditionally, ketamine was analyzed with gas chromatography (GC) equipped with nitrogen-phosphorus detection, flame-ionization detection, and mass selective detection (MSD) or with liquid chromatography-mass spectrometry (LC-MS). These procedures are sensitive but tedious and slow. There is no commercial immunoassay for ketamine. We have developed a simple and rapid electrospray ionization MS (ESI-MS) procedure to screen ketamine and norketamine (NK) in urine samples. Samples were spiked with ketamine-d(4) (K-d(4)) as internal standard and extracted with 0.2 mL of hexane. An experienced technician can prepare a batch of 60 samples in 1 h. An Agilent LC-MSD trap system with autosampler was employed to inject 10-microL extracted samples directly for mass analysis without chromatographic separation. Total analysis time was 1.3 min per sample. The ESI MS was operated in scan mode. The ion pairs (m/z 238/242 for K/K-d(4) and m/z 224/242 for NK/K-d(4)) extracted from the full scan mass spectrum were used for quantification. Because of the nature of the ion trap mass detector employed, the presence of other compounds at high concentration could cause the suppression of target analyte ion intensity determined. Limits of detection were 3 ng/mL for ketamine and 15 ng/mL for NK. Carryover was 0.28% for ketamine and 0.39% for norketamine. Within-run precision (%CV) for K and NK at 3 different concentrations (80, 200, and 600 ng/mL) was 4.0% to 14.7%. A group of 168 urine samples collected from disco-dancing club participants were screened with ESI-MS and confirmed with GC-MS. The sensitivity was 97.1% and specificity was 85.7%. These results indicated that the ESI-MS screening procedure is rapid, sensitive, accurate, and reliable. PMID- 15538965 TI - A study of the effectiveness of commercially available drink test coasters for the detection of "date rape" drugs in beverages. AB - The use of illicit substances for the purpose of drug-facilitated sexual assault (DFSA) poses a significant problem. There has been an increase in public awareness of this problem, and a recent invention in the form of a drink coaster claims to detect whether or not a beverage has been spiked with a so-called date rape drug. A person is instructed to place a drop of the suspect beverage onto two spots of the test, smear gently, and wait until dry. If either spot turns to a darker blue color, then a possible date rape drug has been detected by the coaster test. In an effort to determine the effectiveness of the coasters, various drugs that have been associated with drug-facilitated sexual assault were tested at different concentrations in a variety of common alcoholic and non alcoholic beverages. It was found that although the coasters do detect the presence of GHB and ketamine, two drugs that have been associated with DFSA, there are limiting factors such as the high concentration of the drugs required, hindrance of the reaction due to beverage matrix, and extensive time requirements for ketamine analysis. PMID- 15538966 TI - A sertraline-intoxicated driver. AB - Sertraline is a selective serotonin reuptake inhibitor (SSRI) that is chemically unrelated to other SSRIs, tricyclic antidepressants, and other currently available antidepressant medications. This report documents a case of driving under the influence of sertraline. The subject was involved in a motor vehicle accident. Upon contact by law enforcement, the subject was confused and could neither stand nor walk. The officer noted mumbled speech, droopy eyes, and that the subject seemed sleepy. No alcohol was present in the vehicle, and no odor of alcohol was detected on the subject's breath. The subject was determined to be under the influence of some intoxicating substance. Toxicological analysis revealed only the presence of sertraline. Sertraline was extracted from the blood sample utilizing solid-phase extraction and identified and quantitated by gas chromatography-mass spectrometry. The blood sertraline concentration was determined to be 1285 microg/L. PMID- 15538967 TI - Biomarkers of neurodegenerative disorders: how good are they? AB - Biomarkers are very important indicators of normal and abnormal biological processes. Specific changes in pathologies, biochemistries and genetics can give us comprehensive information regarding the nature of any particular disease. A good biomarker should be precise and reliable, distinguishable between normal and interested disease, and differential between different diseases. It is believed that biomarkers have great potential in predicting chances for diseases, aiding in early diagnosis, and setting standards for the development of new remedies to treat diseases. New technologies have enabled scientists to identify biomarkers of several different neurodegenerative diseases. The followings, for instance, are only a few of the many new biomarkers that have been recently identified: the phosphorylated tau protein and aggregated Beta-amyloid peptide for Alzheimer's disease (AD), Alpha-synuclein contained Lewy bodies and altered dopamine transporter (DAT) imaging for Parkinson's disease (PD), SOD mutations for familial amyotrophic lateral sclerosis (ALS), and CAG repeats resulted from Huntington's gene mutations in Huntington's disease (HD). This article will focus on the most-recent findings of biomarkers belonging to the four mentioned neurodegenerative diseases. PMID- 15538968 TI - Following a TRAIL: update on a ligand and its five receptors. AB - Identification of tumour necrosis factor apoptosis inducing ligand (TRAIL), a TNF family ligand, sparked a torrent of research, following an initial observation that it could kill tumour cells, but spare normal cells. Almost a decade after its discovery, and with five known receptors, the true physiological role of TRAIL is still debated and its anti-tumorigenic properties limited by potential toxicity. This review takes a comprehensive look at the story of this enigmatic ligand, addressing its remaining potential as a therapeutic and providing an overview of the TRAIL receptors themselves. PMID- 15538969 TI - Defective maintenance of intracellular Ca2+ homeostasis is linked to increased muscle fatigability in the MG29 null mice. AB - Mitsugumin 29 (MG29) is a transmembrane protein that is normally found in the triad junction of skeletal muscle. Our previous studies have shown that targeted deletion of mg29 from the skeletal muscle resulted in abnormality of the triad junction structure, and also increased susceptibility to muscle fatigue. To elucidate the basis of these effects, we investigated the properties of Ca2+ uptake and -release in toxin-skinned Extensor Digitorium Longus (EDL) muscle fibers from control and mg29 knockout mice. Compared with the control muscle, submaximal Ca2+-uptake into the sarcoplasmic reticulum (SR) was slower and the storage of Ca2+ inside the SR was less in the mutant muscle, due to increased leakage process of Ca2+ movement across the SR. The leakage pathway is associated with the increased sensitivity of Ca2+/caffeine -induced Ca2+ release to myoplasmic Ca2+. Therefore, the increased fatigability of mutant EDL muscles can result from a combination of a slowing of Ca2+ uptake, modification of Ca2+ induced Ca2+ release (CICR), and a reduction in total SR Ca2+ content. PMID- 15538970 TI - Spontaneous Ca2+ oscillations in subcellular compartments of vascular smooth muscle cells rely on different Ca2+ pools. AB - Spontaneous Ca2+ oscillations in vascular smooth muscle cells have been modeled using a single Ca2+ pool. This report describes spontaneous Ca2+ oscillations dependent on two separate Ca2+ sources for the nuclear versus cytoplasmic compartments. Changes in free intracellular Ca2+ were monitored with ratiometric Ca2+- fluorophores using confocal microscopy. On average, spontaneous oscillations developed in 79% of rat aortic smooth muscle cells that were synchronous between the cytoplasm and nucleus. Reduction of extracellular Ca2+ (less than 1 microM)decreased the frequency and amplitude of the cytoplasmic oscillations with 48% of the oscillations asynchronous between the nuclear and cytoplasmic compartments. Similar results were obtained with the Ca2+ channel blockers, nimodipine and diltiazem. Arg-vasopressin (AVP) induced a rapid release of intracellular Ca2+ stores that was greater in the nuclear compartment (4.20 +/ 0.23 ratio units, n = 56) than cytoplasm (2.54 +/- 0.28) in cells that had spontaneously developed prior oscillations. Conversely, cells in the same conditions lacking oscillations had a greater AVP-induced Ca2+ transient in the cytoplasm (4.99 +/- 0.66, n = 17) than in the nucleus (2.67 +/- 0.29). Pre treatment with Ca2+ channel blockers depressed the AVP responses in both compartments with the cytoplasmic Ca2+ most diminished. Depletion of internal Ca2+ stores prior to AVP exposure blunted the nuclear response, mimicking the response of cells that lacked prior oscillations. Spontaneous oscillating cells had a greater sarcoplasmic reticulum network than cells that did not oscillate. We propose that spontaneous nuclear oscillations rely on perinuclear sarcoplasmic reticulum stores, while the cytoplasmic oscillations rely on Ca2+ influx. PMID- 15538971 TI - Overexpression of the tissue inhibitor of metalloproteinase-3 during Xenopus embryogenesis affects head and axial tissue formation. AB - Tissue inhibitors of metalloproteinases (TIMPs) modulate extracellular matrix remodeling during embryonic development and disease. TIMP-3 expression was examined during Xenopus laevis embryogenesis: TIMP-3 transcripts detected in the maternal pool of RNA increased at the mid-blastula transition, decreased dramatically during gastrulation and increased again during neurulation and axis elongation. Interestingly, the decrease during gastrulation was not seen in LiCl treated (dorsalized) embryos. Whole mount in situ hybridization of TIMP-3 using DIG-labeled RNA probes demonstrated that the transcripts were present in all dorsal tissues during embryogenesis, but were prominent only in head structures starting at stage 35. Overexpression of TIMP-3 through transgenesis and RNA injections led to developmental abnormalities and death. Both overexpression strategies resulted in post-gastrulation perturbation including those to neural and head structures, as well as truncated axes. However, RNA injections resulted in more severe early defects such as failure of neural tube closure, and transgenesis caused truncated axes and head abnormalities. No transgenic embryo expressing TIMP-3 survived past stage 40. PMID- 15538972 TI - Soluble expression and characterization of a GFP-fused pea actin isoform (PEAc1). AB - A pea actin isoform PEAc1 with green fluorescent protein (GFP) fusion to its C terminus and His-tag to its N-terminus, was expressed in prokaryotic cells in soluble form, and highly purified with Ni-Chelating Sepharose Fast Flow column. The purified fusion protein (PEAc1-GFP) efficiently inhibited DNase I activities before polymerization, and activated the myosin Mg-ATPase activities after polymerization. The PEAc1-GFP also polymerized into green fluorescent filamentous structures with a critical concentration of 0.75 uM. These filamentous structures were labeled by TRITC-phalloidin, a specific agent for staining actin microfilaments, and identified as having 9 nm diameters by negative staining. These results indicated that PEAc1 preserved the essential characteristics of actin even with His-tag and GFP fusion, suggesting a promising potential to use GFP fusion protein in obtaining soluble plant actin isoform to analyze its physical and biochemical properties in vitro. The PEAc1-GFP was also expressed in tobacco BY2 cells, which offers a new pathway for further studying its distribution and function in vivo. PMID- 15538973 TI - Ectopic expression of clusterin/apolipoprotein J or Bcl-2 decreases the sensitivity of HaCaT cells to toxic effects of ropivacaine. AB - Local anesthetics inhibit cell proliferation and induce apoptosis in various cell types. Ropivacaine, a unique, novel tertiary amine-type anesthetic, was shown to inhibit the proliferation of several cell types including keratinocytes. We found that Ropivacaine could inhibit the proliferation and induce apoptosis in an immortalized human keratinocyte line, HaCaT, in a dose- and time-dependent manner and with the deprivation of serum. The dose-dependent induction of apoptosis by ropivacaine was demonstrated by DNA fragmentation analysis and the proteolytic cleavage of a caspase-3 substrate-poly (ADP-ribose) polymerase (PARP). In addition, ropivacaine downregulated the expression of clusterin/ apoliporotein J, a protein with anti-apoptotic properties, in a dose-dependent manner, which well correlated with the induction of apoptosis of HaCaT cells. To investigate the role of clusterin/apoliporotein J in ropivacaine-induced apoptosis, HaCaT cells overexpressing clusterin/apoliporotein J were generated and compared to cells expressing the well established anti-apoptotic Bcl-2 protein. Ectopic overexpression of the secreted form of clusterin/apoliporotein J or Bcl-2 decreased the sensitivity of HaCaT cells to toxic effects of ropivacaine as demonstrated by DNA fragmentation, the proteolytic cleavage of PARP and by a reduction in procaspase-3 expression. Furthermore, the downregulation of endogenous clusterin/apolipoprotein J levels by ropivacaine suggested that this might be one mechanism by which ropivacaine induced cell death in HaCaT cells. In conclusion, the ability of ropivacaine to induce antiproliferative responses and to suppress the expression of the anti-apoptotic protein clusterin/apolipoprotein J, combined with previously reported anti-inflammatory activity and analgesic property of the drug, suggests that ropivacaine may have potential utility in the local treatment of tumors. PMID- 15538974 TI - The expression of core fucosylated E-cadherin in cancer cells and lung cancer patients: prognostic implications. AB - It is well documented that the glycosylation of E-cadherin is correlated with cancer metastasis, but whether E-cadherin could be core fucosylated remains largely unknown. We found that E-cadherin was core fucosylated in highly metastatic lung cancer cells while absent in lowly metastatic lung cancer cells. Since alpha-1,6 Fucosyltransferase (alpha-1,6 FucT) is known to catalyze the reaction of core fucosylation, we investigated the biological function of core fucosylation on E-cadherin by alpha-1,6 FucT targeted RNAi and transfecting alpha 1,6 FucT expression vector. As a result, calcium dependent cell-cell adhesion mediated by E-cadherin was strengthened with the reduction of core fucosylation on E-cadherin after RNAi and was weakened with the elevated core fucosylation on E-cadherin after alpha-1,6 FucT over expression. Our data indicated that alpha 1,6 FucT could regulate E-cadherin mediated cell adhesion and thus play an important role in cancer development and progression. Computer modeling showed that core fucosylation on E-cadherin could significantly impair three-dimensional conformation of N-glycan on E-cadherin and produce conformational asymmetry so as to suppress the function of E-cadherin. Furthermore, the relationship between the expression of core fucosylated E-cadherin and clinicopathological background of lung cancer patients was explored in lung cancer tissue of patients. It turns out to demonstrate that core fucosylated E-cadherin could serve as a promising prognostic indicator for lung cancer patients. PMID- 15538975 TI - The effect of C-terminal fragment of JNK2 on the stability of p53 and cell proliferation. AB - The basal activity of JNK is low in normal growing cells and inactivated JNK targets p53 for ubiquitination. To elucidate if the C-terminal part of JNK is responsible for its binding to p53, the low background tet-off inducible NIH3T3 cell line was selected by luciferase reporter gene and a double stable C-JNK Aa (203-424) cell line was established. After withdrawing tetracycline, the C-JNK fragment expression was induced and cell growth was dramatically inhibited 24 h later. However, the expression of p53 was found to be increased after the induction of C-JNK fragment, evaluated by transfecting p21waf-luciferase reporter genes. Our further studies showed that C-JNK fragment could form complex with p53 both in vivo and in vitro. Induction of C-JNK fragment in vivo can increase p53 stability by inhibiting p53 ubiquitination. PMID- 15538976 TI - A new trick to tune down TGF-beta signal. AB - Signal transduction in early embryogenesis needs to be properly controlled. A new player involved in tuning down TGF-beta signaling has now been identified - new evidence that multi-layer control of signaling is essential in vivo. PMID- 15538977 TI - Eradicating Nasal Carriage of Staphylococcus aureus As a Strategy for Preventing Health Care-associated Infections in Nonsurgical Patients. PMID- 15538978 TI - The Role of the Intestinal Tract As a Source for Transmission of Nosocomial Pathogens. AB - The intestinal tract provides an important source for transmission of many nosocomial pathogens, including Enterococcus species, Clostridium difficile, Candida species, Enterobacteriaceae, and other gram-negative bacilli. Recent data suggest that the intestinal tracts of hospitalized patients may also be an important reservoir of Staphylococcus aureus. Although the clinical manifestations of these pathogens are diverse, a common pathogenesis is involved in their colonization of and dissemination from the intestinal tract. Of particular importance is the role that antibiotic selective pressure plays in promotion of colonization by antibiotic-resistant pathogens. Strategies to limit the spread of these pathogens must include efforts to improve adherence to standard infection control practices and promotion of good antimicrobial stewardship. New strategies that include application of novel technologies to the problem of pathogen transmission are needed, and additional research is needed to clarify the potential utility of selective decontamination of the digestive tract. PMID- 15538979 TI - Current Strategies for Prevention of Surgical Site Infections. AB - Surgical site infections (SSIs) are a common complication that follows all types of operative procedures. These infections are usually caused by the exogenous and endogenous microorganisms that enter the operative wound during the course of surgery. The general and procedure-specific risk factors for the development of SSI have been identified and are discussed in this article. Factors that influence the SSI rate and the current strategies for prevention of SSIs are also presented. Emphasis is placed on the efficacious use of antibiotic prophylaxis in surgery. A discussion of the principles of antibiotic prophylaxis, including choice of agents, route of administration, and timing, is offered. It appears that the use of less invasive laparoscopic surgical approaches, as practiced widely today, will be associated with an overall decreased incidence of SSI. PMID- 15538980 TI - Staphylococcus aureus Colonization and Nosocomial Infections: Implications for Prevention. AB - Colonization with Staphylococcus aureus is a risk factor for invasive S. aureus infections. Intranasal mupirocin has effectively eradicated S. aureus colonization and appears to prevent S. aureus nosocomial infections in some patient populations. In hospitals and communities where mupirocin use has been widespread, resistance to the drug has emerged. New strategies and agents are needed if we want to significantly decrease the risk of S. aureus infections. PMID- 15538981 TI - Candida Vulvovaginitis in Pregnancy. AB - In the absence of epidemiologic studies over many decades, there are scant data on the prevalence and natural history of vaginal colonization and symptomatic Candida infections in pregnant women. The hormonal mileau of the vagina during pregnancy undoubtedly enhances Candida colonization and serves as a risk factor for symptomatic expression; however, the frequency, clinical manifestation, and response to therapy is largely based on empiric diagnosis of unsubstantiated disease. For reasons of liability, pregnant women have been largely excluded from controlled clinical treatment studies by the pharmaceutical industry; accordingly, treatment guidelines are dictated by opinions rather than data. As in nongravid women, management of symptomatic pregnant women should be based on confirmed diagnosis and not clinical impression. PMID- 15538982 TI - Vaginal Immunity in Bacterial Vaginosis. AB - Vaginal immunity in response to microbial perturbation is still poorly understood and may be crucial for protection from adverse outcomes associated with bacterial vaginosis (BV). BV is the most prevalent vaginal disorder in adult women worldwide. However, its pathogenesis is still elusive. In BV-positive women, inflammatory signs are scant--approximately 50% of women are asymptomatic. The number of vaginal neutrophils in the BV-positive patient is not increased with respect to healthy women. In contrast, vaginal interleukin (IL)-1beta levels are largely increased. Recent findings indicate that microbial hydrolytic enzymes could be responsible for dampening the expected proinflammatory response cascade after IL-1beta increase. In other words, BV causes a large increase of vaginal IL 1beta, which is not paralleled by an increase of IL-8 levels, suggesting that BV associated factors specifically dampen IL-8. The impairment of IL-8 increase may explain the absence of neutrophil increase in most women exposed to a massive abnormal anaerobic vaginal colonization (BV). Among BV-positive women, vaginal innate immunity is strongly correlated to a specific adaptive immune response: the immunoglobulin A (IgA) against the hemolysin produced by Gardnerella vaginalis (anti-Gvh IgA), which is the main bacterium present in BV. High anti Gvh IgA levels are protective for adverse pregnancy outcomes. However, an exaggerated inflammatory response, mainly attributed to genetic polymorphisms, is also implicated in BV-associated adverse outcomes. PMID- 15538983 TI - Antimicrobial Selection in the Treatment of Pyelonephritis. AB - Pyelonephritis is a common clinical entity, although accurate data regarding the true prevalence of this infection are lacking. Acute pyelonephritis is associated with significant morbidity and even mortality. There are very few randomized controlled clinical trials that have addressed the optimal management strategies and antimicrobial therapy for this infection. Increasing resistance among uropathogens, especially Escherichia coli, has impacted recommendations for empiric antimicrobial therapy, and fluoroquinolones have emerged as the empiric therapy of choice for individuals managed as outpatients; more options are available for empiric parenteral therapy for those who require hospitalization. Further study of the epidemiology of antimicrobial resistance among uropathogens and of alternative agents for the management of pyelonephritis is urgently needed. PMID- 15538984 TI - Innate Immune System Gene Polymorphisms in Women with Vulvovaginal Infections. AB - The innate immune responses at mucosal surfaces of the lower female genital tract play a central role in preventing the establishment of infection. Variability in the genes regulating innate immune recognition or response to infectious microorganisms could explain interindividual differences in susceptibility to infection and severity of infectious disorders. Of the numerous genetic variations identified within immunoregulatory genes, only a few have so far been studied in relation to infectious disorders of the lower female genital tract. Although these studies broaden our understanding of the genetic influence on mucosal innate immunity and microbiologic outcome, they also highlight the complexity of links between genotypical and phenotypical features, ie, the influence of a single genetic marker on the phenotype is at best moderate and is not constant in every ethnic/racial group. Such inconsistency is primarily attributed to gene-to-gene interactions and demographic genetic variability. Large case-control studies evaluating multiple genetic markers simultaneously in well-defined subgroups will characterize patients more accurately and pave the way to personalized medicine. PMID- 15538985 TI - The Effects of HIV Protease Inhibitors on Carbohydrate and Lipid Metabolism. AB - Since the introduction of HIV protease inhibitors (PIs), disorders of glucose and lipid metabolism have emerged. In dissecting out the direct effect on lipid and glucose metabolism, it has become apparent that individual PIs have different effects on metabolism. Some PIs such as indinavir acutely induce insulin resistance. PIs have also been shown to cause other disorders of glucose metabolism, including impairment of insulin secretion and increased endogenous glucose production. Individual PIs also have different effects on lipid metabolism. Ritonavir predominantly increases triglyceride and very low-density lipoprotein cholesterol levels. Limited studies in HIV-negative volunteers suggest that several of the PIs do not increase low-density lipoprotein cholesterol levels. This review examines the direct effects of PIs on glucose and lipid metabolism by assessing prospective studies of HIV-infected and healthy normal volunteers, and in vitro studies. PMID- 15538987 TI - Diagnosing and treating depression earlier and preventing recurrences: still neglected after all these years. PMID- 15538986 TI - Illness of Immune Reconstitution: Recognition and Management. AB - Some individuals who initiate highly active antiretroviral therapy (HAART) develop new or worsening opportunistic infections or malignancies despite improvements in surrogate markers of HIV-1 infection. These events of paradoxical clinical worsening, also known as immune reconstitution syndromes (IRS), are increased in individuals with prior opportunistic infections or low CD4+ T-cell nadirs. They are thought to result from reconstitution of the immune system's ability to recognize pathogens or tumor antigens that were previously present, but clinically asymptomatic. There is no consensus regarding the diagnostic criteria or pathogenesis of IRS. Knowledge of their presentation and treatment is largely based on case reports. With the introduction of HAART into resource limited settings, it is likely that significantly more and distinct forms of IRS will be observed. Prospective studies of the incidence and treatment of IRS in multiple settings are critical to better understand their pathogenesis and optimal management. PMID- 15538989 TI - The effectiveness of atypical antipsychotic medications in depressive disorders. AB - Clinical evidence supporting the use of atypical antipsychotic medication (broad spectrum psychotropic agents) in the treatment of depressive disorders is increasing rapidly. Animal models suggest that when atypical antipsychotic medications are used in combination with a selective serotonin reuptake inhibitor there is additional activation of frontal dopaminergic and noradrenergic neurotransmitter systems. This stimulated the initiation of several clinical trials that showed the efficacy of atypical antipsychotic medication augmentation of selective serotonin reuptake inhibitors for patients with treatment-resistant depression. There also are few case reports of successful treatment of depression with atypical antipsychotic medication alone. When a clinician is treating a depressed patient who did not achieve relief after trials with two different antidepressant regimens, one option to consider is augmentation with an atypical antipsychotic medication to ameliorate depressive symptoms. PMID- 15538988 TI - The epidemiology of mood disorders. AB - This review provides an overview of the epidemiology, risk factors, and genetic epidemiology of mood disorders in adults and children. The magnitude and impact of mood disorders in the community outweighs that of most other chronic diseases. Although there is substantial knowledge regarding the sociodemographic risk factors for mood disorders, our understanding of the pathogenesis and classification still is evolving. Comorbidity of mood disorders with anxiety disorders and substance abuse has been documented widely. Whereas substance abuse and mood disorders seem to be independent etiologically, anxiety and mood disorders result from partially common etiologic factors. The results of family, twin, and adoption studies reveal that a positive family history is the most potent risk factor for mood disorders, particularly bipolar disorder. However, the specific factors that are transmitted in families still are unknown. The two areas that will inform future genetic research include phenomenologic studies that refine the validity of the current phenotypic classification of mood disorders, and application of study designs to elucidate specific factors that may explain the familial transmission of these disorders. PMID- 15538990 TI - Nutrients, neurodevelopment, and mood. AB - Human neurodevelopment is the result of genetic and environmental interactions. This paper examines the role of prenatal nutrition relative to psychiatric disorders and explores the relationship among nutrients, mood changes, and mood disorders. Epidemiologic studies have found that adults who were born with a normal, yet low birth weight have an increased susceptibility to diseases such as coronary heart disease, diabetes, and stroke in adulthood. Prenatal caloric malnutrition, low birth weight, and prematurity also increase the risk for neurodevelopmental disorders, schizophrenia, affective disorders, and schizoid and antisocial personality disorders. Placebo-controlled studies in medicated patients suggest that add-on treatment with omega-3 fatty acids, particularly eicosapentaenoic acid, may ameliorate symptoms of major depressive disorder. Additional studies are necessary to confirm any benefits for bipolar disorders. PMID- 15538992 TI - Quality of life in patients with coronary artery disease and the impact of depression. AB - Depression has been shown to be a risk factor for the development of coronary artery disease, and also is associated with greater risk of mortality in patients with coronary disease. Treatment of depression may or may not prove to alter depression as a risk factor, but improving the quality of life (QOL) of patients with coronary disease also should be considered a primary outcome. Quality of life is a difficult concept to define and measure. However, recent investigations in patients with coronary disease have examined the relationship of QOL to mortality risk and depression. This article will review the concept and measurement of QOL, discuss studies of factors that predict QOL in patients with coronary artery disease, and focus on the impact of depression on QOL. PMID- 15538993 TI - Issues of functionality in bipolar disorders. AB - This is a review derived from the abstracts of a group of symposia and posters presented at the International Society for Bipolar Disorders Regional Group Conference, held in Sydney, Australia, February 5-7, 2004. It consists of selected abstracts published collectively. In part due to sponsorship by the pharmaceutical industry, information presented at national and international meetings often predominantly involves the use of psychotropic drugs. In contrast, the current summary of studies focuses on the impact of bipolar disorders on functionality as this relates to such variables as self-satisfaction, the ability to work, and the ability to get along interpersonally. PMID- 15538991 TI - Increased mortality in depressive disorders: a review. AB - Several factors have been proposed to explain the relationship between excess mortality and depressive disorders. These include mechanisms such as increased suicide rates, hazardous health behavior (smoking, alcohol use, unhealthy eating), psychologic reactions to developing a medical illness, biological dysregulations (hyperactivity of the hypothalamic pituitary adrenal, neuro-immune dysregulation, sympathoadrenergic dysregulation), and noncompliance with medical treatment. The evidence supporting the role of each of these mechanisms in excess mortality varies considerably. The causal direction in most of the mechanisms is not clear. It is possible that the explanatory factors, such as smoking, compliance, or biological mechanisms, cause depression, or that depression causes these factors, or that both are explained by a third, underlying factor. We will summarize the evidence supporting these mechanisms, and propose options for possible interventions aimed at reducing the increased risk of dying. PMID- 15538995 TI - Separate and concomitant use of lamotrigine, lithium, and divalproex in bipolar disorders. AB - Expert consensus emphasizes the need for better recognition and accurate diagnosis of bipolar disorder. Current research on lithium, divalproex, and lamotrigine provides new insight into the effective management of this illness. Advances in identifying the mechanism of action of mood stabilization has focused on signaling pathways within the cell that are associated with neurotrophic effects. Clinical research has led to confirmatory evidence of the efficacy of lithium in all phases of bipolar disorder, with the greatest effects seen in the treatment and prevention of mania. Compared to divalproex, lithium also has been found to have greater efficacy in the prevention of suicide. Lamotrigine has emerged as a first line treatment for bipolar depression, which is an area of weakness for other mood stabilizers. Oral loading of divalproex leads to rapid stabilization of mania without imposing a greater adverse effect burden than conventional dosing. Because no agent is universally effective in all phases of the illness, combination therapy with two or more agents often is the best option. PMID- 15538994 TI - Cognition in mania and depression: psychological models and clinical implications. AB - Affective disorders, including bipolar disorder and major depressive disorder, are highly prevalent throughout the world and are extremely disabling. Diagnostic and Statistical Manual criteria and psychological models strongly implicate cognitive dysfunctions as being integral to our understanding of these disorders. We review the findings from studies that have used neurocognitive tests and functional imaging techniques to explore abnormal cognition in affective disorders. In particular, we highlight the evidence for cognitive dysfunctions that persist into full clinical remission, and the recent trend toward the use of "hot" processing tasks, involving emotionally charged stimuli, as a means of differentiating between the cognitive underpinnings of mania and depression. The clinical relevance of these developments is discussed. PMID- 15538996 TI - Neurochemical predictors of response to pharmacologic treatments for bipolar disorder. AB - Bipolar disorder is a common psychiatric disorder that is characterized by recurrent episodes of mania and often depression. Mood stabilizers and antipsychotics are first-line pharmacologic options for patients with bipolar disorder. However, the exact mechanisms by which these medications exert the mood stabilizing effects are unknown. Additionally, individuals with bipolar disorder often try several medications unsuccessfully before achieving mood stabilization. Magnetic resonance spectroscopy (MRS) is a noninvasive neuroimaging technique that can be used to identify the neurochemical effects and predictors of response to medications commonly used to treat bipolar disorder. MRS may facilitate targeted treatment interventions and decrease the morbidity and mortality associated with this illness. Examining the mechanisms of action of pharmacologic agents used to treat bipolar disorder may clarify the neurophysiologic basis of bipolar disorder. We will review recent MRS investigations that have evaluated the neurochemical effects of pharmacologic treatments and predictors of treatment response in patients with bipolar disorder. PMID- 15538997 TI - Relationship of acute mania symptomatology to maintenance treatment response. AB - Most treatment of bipolar disorders addresses maintenance objectives. In the past 4 years, several large, blinded, randomized, placebo-controlled maintenance studies involving more than 3000 patients with bipolar I illness have been published, with analyses of background, symptomatic, and acute treatment factors contributing to maintenance effectiveness. This article summarizes these findings. Generally, indices of greater severity predict lower response rates to most monotherapy treatments. Some findings have been unexpected. Mixed mania did not predict different maintenance response to divalproex or lithium but predicted more side effects with either drug, and reduced efficacy of olanzapine or lamotrigine. PMID- 15538999 TI - The metabolic syndrome and type 2 diabetes: what has gone wrong in type 2 diabetes care? PMID- 15538998 TI - Treatment of seasonal affective disorder: unipolar versus bipolar differences. AB - Evidence-based treatments for seasonal affective disorder (SAD) include light therapy and pharmacotherapy. We briefly review the diagnosis and treatment of SAD, focusing on clinical and treatment differences between patients with unipolar and bipolar illness. Special considerations for the management of SAD in patients with bipolar disorder are discussed, including the need to monitor for emergence of manic and hypomanic mood switches, to use mood stabilizers in patients with bipolar I disorder, and to be aware of potential interactions between bright light and medications used in treating bipolar disorder. Chronobiological treatments such as bright light therapy may be combined with pharmacotherapy to enhance therapeutic effects, reduce adverse side effects, and optimize treatment in patients with seasonal and nonseasonal bipolar disorder. PMID- 15539002 TI - Aldose reductase inhibition in the treatment of diabetic neuropathy: where are we in 2004? AB - Diabetic neuropathy is a major complication of poorly controlled diabetes mellitus. Aldose reductase, the first enzyme of the polyol pathway, is thought to play a role in initiating the metabolic damage to peripheral nerves during hyperglycemia. Aldose reductase inhibitors (ARIs) have been proposed to dampen the flux of glucose through the pathway during hyperglycemia; however, clinical trials in diabetic patients to demonstrate efficacy in the prevention or amelioration of diabetic neuropathy have failed thus far. Recent improved understanding of the pitfalls of past trials and some improved ARIs and clinical evaluation instruments show promise that success in the 20-plus year search for efficacious ARIs may soon be at hand. PMID- 15539003 TI - Chronic inflammatory demyelinating polyradiculoneuropathy in diabetes mellitus. AB - There is a higher incidence of demyelinating peripheral neuropathy responsive to immunomodulating treatment in patients with diabetes mellitus. The diagnosis is often overlooked and the patients are given the label of "diabetic neuropathy." Progressive symmetric or asymmetric motor deficit, progressive sensory neuropathy in spite of optimal diabetic control, and unusually high cerebrospinal fluid protein level in "diabetic neuropathy" should alert the clinician to the possibility of an underlying treatable demyelinating peripheral neuropathy masquerading as "diabetic neuropathy." PMID- 15539004 TI - Reducing the incidence of foot ulceration and amputation in diabetes. AB - Diabetes is a common disease that is associated with numerous complications, including foot ulceration and amputation. In diabetic patients, the incidence of foot ulcers ranges from 1.0% to 4.1%, and the incidence of lower-extremity amputations ranges from 2.1 to 13.7 per 1000. Risk factors for developing foot ulcers and subsequent amputation include neuropathy, peripheral vascular disease, and trauma. To reduce these complications, several preventive strategies have been devised, from reducing risk factors to improving treatment and management. PMID- 15539005 TI - Research development in the pathogenesis of neuropathic diabetic foot ulceration. AB - Neuropathic ulceration is the most prevalent type of chronic lesion of the diabetic foot. Its pathogenesis is mainly related to the abnormally increased pressure on the sole of the foot, secondary to lack of sensation and deformities induced by peripheral sensory-motor neuropathy, which are associated with an abnormal gait. The incidence of the ulcerative state is brought on by repetitive postural trauma, which causes a chronic inflammatory state, characterized by hypertrophy of skin layers and diffuse, intense inflammatory infiltrate throughout the ulcer. The off-loading of the ulcer will restore the healing process, leading to a reparative phase that will be histologically and biochemically evident. PMID- 15539006 TI - Is this bone infected or not? Differentiating neuro-osteoarthropathy from osteomyelitis in the diabetic foot. AB - Osteomyelitis (bone infection) and neuro-osteoarthropathy (Charcot arthropathy) are limb-threatening complications of diabetic neuropathy with very different therapies. Distinguishing between them may be difficult, but it is important. In Charcot arthropathy, noninfectious soft tissue inflammation accompanies rapidly progressive destruction, first of joints, then of bone. This occurs in a well vascularized and severely neuropathic, but nonulcerated, foot. In osteomyelitis, chronic soft tissue ulceration precedes infection of bone, which may be physically exposed. Magnetic resonance imaging and bone biopsy are the preferred diagnostic tests, provided adequate technical and interpretive skills are available. PMID- 15539007 TI - Intensive treatment of diabetic nephropathy. PMID- 15539008 TI - Use of genetic mouse models in the study of diabetic nephropathy. AB - The study of experimental diabetic nephropathy in rodent models has led to many changes in the clinical management of human diabetic nephropathy. With the development of technology to generate knockout and transgenic animals, the mouse has become a favored species in medical research. There are several genetic mouse models of diabetes, with the majority being models of type 2 diabetes mellitus. These include the hypoinsulinemic nonobese diabetic mouse, the KKAy mouse, the New Zealand obese mouse, the hyperinsulinemic ob/ob mouse, and the different strains of obese hyperinsulinemic db/db mouse. Each of these models displays some renal changes, but by far the best model of renal disease and the one that is the most studied is the db/db mouse. The db/db mouse displays substantial glomerular pathology, including mesangial matrix expansion and modest albuminuria. It has been reported that the db/db mouse has a decline in creatinine clearance after 5 months of age, but more specific approaches are warranted to confirm these findings. A number of intervention studies show renoprotection in this model. Although mice have many advantages, such as being able to be crossbred with genetically manipulated animals, in many ways they are not very similar to humans, and in some respects the rat may be a better choice, particularly in relation to some features of end-organ injury. PMID- 15539009 TI - Clinical studies of advanced glycation end product inhibitors and diabetic kidney disease. AB - Current treatment of the nephropathy complication of diabetes mellitus is suboptimal in halting the progression of the complex disease. Among the irreversible effects of sustained hyperglycemia is the heightened formation of advanced glycation end products (AGEs). The role of AGEs in diabetic nephropathy has been established by years of basic research. This article reports progression through human studies of the few AGE inhibitors that have reached clinical development, including pimagedine, pyridoxamine, and alagebrium. PMID- 15539010 TI - Transforming growth factor-beta: a clinical target for the treatment of diabetic nephropathy. AB - Diabetic nephropathy is continuing to rise in incidence, despite awareness of tight glycemic control and blood pressure. The identification that matrix accumulation is driven by transforming growth factor-beta (TGF-beta) has led to a concerted effort to apply antifibrotic strategies for this disorder. Recent studies have not only demonstrated the beneficial effects of blocking TGF-beta on matrix accumulation but have also found that blocking TGF-beta may have important hemodynamic effects that are relevant to diabetic complications. In this article, we review the latest knowledge regarding the role of TGF-beta in diabetic kidney disease and discuss available and novel therapeutic approaches. The role of a novel antifibrotic drug, pirfenidone, may have important clinical relevance to diabetic nephropathy. PMID- 15539011 TI - Diabetic nephropathy in African-American patients. AB - Diabetic nephropathy (DN) is the number one cause of end-stage renal disease (ESRD) in the United States and is highly prevalent in African Americans. Since 1997, DN has been the number one cause of ESRD in African Americans. In African Americans, almost all DN is due to type 2 diabetes mellitus (T2DM), and nephropathy may affect female more than male patients. African Americans with T2DM are at increased risk for developing and having progression of DN. Glycemic control, development of albuminuria, family history of renal disease, and control of blood pressure are important risk factors for progression of DN. In addition, cigarette smoking, presence of hepatitis C, and use of thiazolinediones has an impact on renal survival in African Americans. Large vessel complications may be less frequent in African Americans with T2DM, when compared with white persons. Yet, cardiovascular disease and other microvascular complications are very common, and both are dependent on control of blood pressure. Achieving the recommended blood pressure of less than 130/80 mm Hg is essential but requires multiple antihypertensive medications, including an inhibitor of the renin angiotensin system. PMID- 15539013 TI - [Recommendations for enhancing the quality of traditional Chinese medicine clinical research reporting]. AB - Evidence based medicine (EBM) has drawn the attention of the medical community around the world and is rapidly becoming the standard in medical research. EBM requires both rigorous clinical trial research design, including adequate randomization and appropriate control, and accurate and thorough reporting of the findings of trials. Despite China's active research program, the majority of traditional Chinese medicine (TCM) clinical research in the Chinese literature fails to meet world standards of clinical research reporting, largely due to insufficient and inadequate reporting. Because of rising world demand for information on TCM research, enhancing the quality of research reporting in Chinese journals is imperative. Quality of reporting is important in all aspects of a manuscript, including the introduction/background, materials and methods, results, and discussion/conclusion sections. TCM journal editors, who serve as gatekeepers of quality, must encourage higher quality clinical research reporting by setting and upholding publication standards. PMID- 15539012 TI - Gene expression in diabetic nephropathy. AB - Diabetic nephropathy (DN) is a common complication of diabetes types 1 and 2. One of the hallmarks of DN is the development of mesangial expansion, which occurs through accumulation of extracellular matrix (ECM) components. Altered local gene expression of humoral factors (eg, transforming growth factor-b, connective tissue growth factor, and platelet-derived growth factor) can lead to increased production of ECM components (eg, fibronectin and collagen IV) or decreased degradation through matrix metalloproteinases (eg, MMP-1, MMP-2). In recent years, new techniques for examination of gene expression have been developed. Because of their large scale and high-throughput character, it is now possible to examine differential gene expression in a large number of samples. This paper provides an overview of techniques used and results obtained in studies of DN. Newly developed concepts of how altered gene expression may affect histomorphologic features or clinical symptoms are also discussed. PMID- 15539014 TI - [Strategy for treating liver fibrosis by integration of the disease and its syndromes in traditional Chinese medicine]. AB - The strategy for treating liver fibrosis in chronic hepatitis patients includes dispelling the etiological factors, inhibiting the inflammatory reaction, decreasing the sedimentation of extracellular matrix, accelerating the degradation of extracellular matrix, improving the microcirculatory and metabolic disturbance, and ameliorating the complicating diseases, etc. Researchers should pay attention to the liver function indexes in chronic liver disease in evaluating the therapeutic effects of anti-fibrosis. Effective etiological treatment should be considered as the first step in treating liver fibrosis in chronic hepatitis patients, and inhibiting the inflammatory reaction is one of the most important tactics for suppressing the development of fibrosis and for decreasing the incidence rate of liver cancer in chronic hepatitis patients. Treatment based on syndrome differentiation, a dynamic therapy aimed at the holistic pathological state, can improve the pathological state of the disease. It is especially important to take the advantages of the integration of traditional Chinese and western medicine in the clinical diagnostic and therapeutic procedure for increasing the therapeutic effect of live fibrosis. PMID- 15539015 TI - [Treatment of intrahepatic cholestasis with integrated traditional Chinese and Western medicine]. AB - Intrahepatic cholestasis results from the disturbance of the structure and function of bile secretory apparatus in hepatic cells. At present, the mechanism of the disease is still not clearly understood. The currently used Western medicine for liver-protective, cholagogic and immuno-suppressive treatment are not effective enough. According to the theory of traditional Chinese medicine, moisture, heat, stasis and toxin are the main factors of the etiology and pathogenesis of the jaundice in patients with intrahepatic cholestasis. Therefore, clearing away heat to drain dampness, cooling blood to remove toxin, activating blood to resolve stasis, soothing liver and invigorating spleen should be adopted in the treatment. In order to increase the therapeutic effects, rational application of traditional Chinese herbs combined with Western medicine should be emphasized in treating the disease under the doctrine of integration of syndrome differentiation and disease differentiation. PMID- 15539016 TI - [Significance of adaptive response to study of traditional Chinese medicine]. AB - Adaptive response is a topic of general interest in medical research. Utilization of adaptive response has become a new treatment strategy for some diseases, the core of which is to stimulate the mechanisms of body internal protection, to improve the adaptive and anti-injury capability, and to maintain the homeostasis of the body. The existence of adaptive response indicates that the body internal protection can be induced and its key mechanism is the inducement of internal protective factors (internal protective proteins). In this article, some connections between adaptive response and the theory of traditional Chinese medicine (TCM) were discussed, which indicated that adaptive response is much beneficial for explaining some theories of TCM such as correspondence between human and nature, strengthening healthy qi to consolidate constitution, acupuncture theory, adaptogen herbs, and treating disease before its onset, etc. Thus the adaptive response is of great significance for explaining scientifically the connotative meaning of the theories of TCM and for promoting the integration of TCM and western medicine. PMID- 15539017 TI - [Clinical study on idiopathic thrombocytopenic purpura treated with Shengxueling Granule]. AB - OBJECTIVE: To observe the clinical effect of Shengxueling Granule (SXLG) in treating idiopathic thrombocytopenic purpura (ITP), and to study its possible mechanism. METHODS: Eighty-six cases of ITP were divided into two groups randomly. Fifty-six cases in the treatment group were treated with SXLG, a traditional Chinese medicine, and 30 cases administered with Western medicine (prednisone) were taken as control. Patients in each group took drugs for three months and were under follow-up observation. RESULTS: In SXLG-treated group, the total effective rate in 3 months was 85.71%, similar to 83.33% in prednisone treated group (P>0.05), while the total effective rate in 6 months in the SXLG treated group was 91.07%, higher than 53.33% of the prednisone-treated group (P<0.01), and no obvious side-effects were observed. The patients' bleeding was alleviated or stopped, and their general condition was improved. And the blood platelet count (BPC) was increased, the platelet associated immunoglobulin (PAIg) and interleukin-4 (IL-4) were markedly dropped, the level of natural killer cells activity (NKCA) increased, and the rate of T lymphocyte subsets gradually returned to normal level. Megakaryocytes tended toward maturation on bone marrow smear after SXLG treatment. All differences above were statistically significant. CONCLUSION: SXLG is an effective and safe medicine for ITP. It can regulate the cellular immunity, inhibit the platelet antibody to reduce the destruction of the platelet and to increase the number of platelet, promote the differentiation and maturation of megakaryocyte, facilitate the production and release of platelet, lower the fragility of capillary, and prevent the hemorrhagic tendency. PMID- 15539018 TI - [Professor Zhang Jing-Ren's experience in treating nephrotic syndrome]. PMID- 15539019 TI - [Clinical effect of escin on patients with cutaneous pruritus caused by blood stasis and wind-dryness]. AB - OBJECTIVE: To observe the clinical effect of escin on patients with cutaneous pruritus caused by blood stasis and wind-dryness and to prove the theory that "wind should be treated by regulating blood disorder, and wind disappears after activating blood" in traditional Chinese medicine (TCM). METHODS: Fifty-one patients were randomly divided into escin-treated group (n=30) and loratadine treated group (n=21). The patients in the escin-treated group were treated with escin for 4 weeks (300 mg, b.i.d.), and the patients in the loratadine-treated group were treated with loratadine (10 mg, q.d.). Symptom score reducing index (SSRI) was used to assess the pruritus degree, lesion range and lesion shape before the treatment, 2 weeks and 4 weeks after the treatment, respectively. RESULTS: Two weeks after the treatment, the effective rate of the escin-treated group was 63.3%, and the effective rate of the loratadine-treated group was 67.0%. Four weeks after the treatment, the effective rates were 86.7% and 80.0% in escin- and loratadine-treated groups respectively. There was no statistical difference in total scores of SSRI in two groups (P>0.05), and the scores of pruritus degree and lesion shape also had no statistical difference (P>0.05), while the score of lesion range of the escin-treated group was lower than that of the loratadine-treated group (P<0.05). CONCLUSION: Escin has satisfactory effect in treating pruritus caused by blood stasis and wind-dryness. The study confirms the TCM theory that "wind should be treated by regulating blood disorder, and wind disappears after activating blood". PMID- 15539020 TI - [Effect of integrated traditional Chinese and Western medicine therapy on progression of acute renal failure in patients with chronic renal insufficiency: a short-term clinical study]. AB - OBJECTIVE: To evaluate the effect of integrated traditional Chinese and Western medicine therapy on the progression of acute renal failure in patients with chronic renal insufficiency. METHODS: Thirty-two patients with chronic renal insufficiency developed acute renal failure recently were treated with Chinese herbs and western drugs intravenously and clysterizing of Chinese herbs liquid for 30 minutes, and the treatment course was 14 days. Assessment of liver and renal function, blood routine, electrolytes and endogenous creatinine clearance rate (Ccr) was performed before and 2 weeks after the treatment. RESULTS: The levels of hemoglobin (HB), white blood cell count (WBC) and serum electrolytes showed no significant changes after the treatment. The levels of blood urea nitrogen (BUN) and serum creatinine (SCr) decreased, while the level of Ccr increased significantly (P<0.05) after the treatment. The total effective rate was 65.6%. CONCLUSION: The integrated traditional Chinese and Western medicine therapy can effectively delay the deterioration of renal function in patients with chronic renal insufficiency accompanied by acute renal failure. PMID- 15539021 TI - [Biomechanical and morphological measurement and analysis of roentgenograph for knee osteoarthritis patients]. AB - OBJECTIVE: To study the etiological factors of knee osteoarthritis and to provide the anatomical basis of corrective therapy for knee joint deformity by comparing the biomechanical differences of the knee joints between the osteoarthritis patients and the normal people. METHODS: The sexes, ages, heights, weights, painful characters and accompanied symptoms were recorded, and the femoral angle, tibial angle, femorotibial angle, joint space angle, and the height and width of condylus medialis and lateralis were measured for 29 selected knee osteoarthritis patients. And the biomechanical data of the patients were compared with those of the 19 normal people. RESULTS: The femoral angle of the osteoarthritis patients was smaller and the joint space angle was greater statistically than that of the normal people, and the ratios of height and width of condylus medialis and condylus lateralis of the osteoarthritis patients were greater than those of the normal people. CONCLUSION: In the patients, the gravity line of the lower limb inclines to the condylus medialis, so the condylus medialis is more vulnerable. And the configuration of the condylus of the patients, which is narrower and longer, is different from that of the normal people. It may be one of the etiological factors of knee osteoarthritis. PMID- 15539022 TI - [Function of MMP/TIMP on airway remodeling of bronchial asthma and treating effects of Zhichuan capsule]. AB - OBJECTIVE: To explore the mechanism of the bronchial asthma and to study the treating effects of Zhichuan Capsule on the airway remodeling of asthmatic model rats. METHODS: The rat model was established by being sensitized and activated with different density of ovalbumin through prolonged and repeated exposure for 8 weeks. The rats were randomly divided into model group, Zhichuan Capsule treated group, dexameson treated group, and Zhichuan Capsule and dexameson treated group. Another group of normal rats were taken as control. General histological changes were observed by hematoxylin and eosin stained sections. Being standardized by internal perimeter (Pi), the wall thickness (d), internal area (Ai), outer area (Ao) and wall area (WA) of the airway were quantified by computer-assisted image analysis system. The express of MMP-9, TIMP-1, Col I, Col III and ColV in the airway were examined by immunocytochemical methods. During the course of airway remodeling, the dynamic changes of model rats were observed at different time points (2, 4, 6 and 8 weeks after the activating). Statistical comparison was performed by ANOVA followed by Fisher LSD test. RESULTS: (1) Histologic examination showed eosinophil infiltration within the airway walls, epithelial damage, excessive mucus in the lumen and edema in the submucosa of the airways in model rats, and that the collagen deposition increased accompanied by increasing of TIMP-1. In the model rats, MMP-9 increased at the time point of 2 weeks, but it decreased in the late stage (8 weeks after activating) of airway remodeling. And the level of TIMP-1 was far higher than MMP-9 at the time point of 8 weeks. (2) Zhichuan Capsule could down-regulate the level of TIMP-1 in the airway wall, as well as the thickness of airway wall and the collagen deposition. And there were progressing effects when it was used together with dexameson. CONCLUSION: (1) The early increase of MMP-9 is a key point to start remodeling; and the increase of TIMP-1 in the late stage, which inhibits collagenase activity, may play an important role in developing airway fibrosis. Imbalance between MMP-9 and TIMP-1 is a marker of airway remodeling. (2) Zhichuan Capsule can decrease the deposition of collagen and suppress the airway remodeling by inhibiting the TIMP 1 expression. PMID- 15539023 TI - [Effect of madecassoside on depression behavior of mice and activities of MAO in different brain regions of rats]. AB - OBJECTIVE: To evaluate the effects of madecassoside (MC) on the depression behavior of mice and the activities of monoamine oxidase (MAO) in different rat brain regions. METHODS: Imipramine as the positive contrast medicine, effects of MC on the depression behavior of mice were observed by forced swimming test and reserpine antagonist test. Moclobemide and pargyline as the positive controlled medicines, the activities of monoamine oxidase-A (MAO-A) and monoamine oxidase-B (MAO-B) in different rat brain regions were determined after intragastric administration of MC in 3 different dosages for 3 days or 21 days. RESULTS: (1) The low, middle and high dosages of MC (i.g.) significantly reduced the immobility time of mice in forced swimming test (P<0.05). (2) MC in dosages of 10 mg/kg and 20 mg/kg prevented the lowering of temperature induced by reserpine (P<0.05), while 40 mg/kg had no significant effects on it (P>0.05). (3) With acute administration (3 days), the low, middle and high dosagey of MC (i.g.) significantly inhibited the activity of MAO-A in hippocampus (P<0.01), and the high dosage significantly inhibited the activity of MAO-A in hypothalamus (P<0.01), while the 3 dosages had no significant effects on the activity of MAO-A in cortex (P>0.05). With chronic administration (21 days), MC in 3 dosages had no significant effects on the activities of MAO-A in cortex and hypothalamus (P>0.05), and the high dosage (40 mg/kg) significantly enhanced the activity of MAO-A in hippocampus (P<0.01). (4) With acute administration, MC in dosages of 10 mg/kg and 20 mg/kg significantly inhibited the activity of MAO-B in cortex (P>0.05), and MC in dosage of 10 mg/kg significantly inhibited the activity of MAO-B in hypothalamus (P<0.05), and MC in dosage of 20 mg/kg significantly enhanced the activity of MAO-B in hippocampus (P<0.01). With chronic administration, MC of 3 dosages produced no significant effects on the activities of MAO-B in 3 different rat brain regions (P>0.05). CONCLUSION: These results support the idea that MC produces antidepressant effects through MAO inhibition in rat brain, which seems stronger with acute administration than chronic administration, while its mechanism remains to be further studied. PMID- 15539024 TI - [Treatment of tobacco dependence with wrist-ankle acupuncture: a report of 30 cases]. PMID- 15539025 TI - [Inhibiting effects of Panax notoginseng extracts on proliferation of GES-1 cells and MNNG-transformed GES-1 cells]. AB - OBJECTIVE: Through cell cultivation, we studied the inhibiting effects of the serum of the dog fed with Panax notoginseng extracts on precancerous gastric cells, trying to find the best time points or periods when the extracts' function was the strongest after administration of the extracts to the dog. METHODS: The experiments adopted eternalized human gastric mucosa epithelium GES-1 cells and MC cells gained from GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso guanidine (MNNG) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at different points of time after feeding the dog with Panax notoginseng extracts for experiment. By means of MTT (3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we examined the inhibiting effects of the serum after culturing the GES-1 and MC cells for 72 hours with different concentration (8%, 4%, 2%) of medicated serum obtained from the dog at different points of time, so as to find that, at which points of time the medicated serum obtained, it would be the most effective. RESULTS: The results showed that the GES-1 and MC cells inhibition rates of medicated serum from the points of 2-hour and 6-hour were the highest, and the culture medium containing 8% of medicated serum from these two points had prominent inhibiting effects on both kinds of cells. The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 2-hour was 70.8% (P<0.01) and that of the MC cells was 45.3% (P<0.01). The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 6-hour was 88.5%(P<0.01) and that of the MC cells was 42.4% (P<0.01). CONCLUSION: The points of time with the strongest inhibiting effects are 2 hours and 6 hours after being fed with Panax notoginseng extracts. At these two points, the serum is most effective in inhibiting the proliferation of GES-1 and MC cells. PMID- 15539026 TI - [Treatment of simple obesity with electro-acupuncture and auricular acupoint pressing: a report of 177 cases]. PMID- 15539027 TI - [A study on anti-inflammatory and analgesic effects of alkaloids of Toddalia asiatica]. AB - OBJECTIVE: To study the pharmacological activities and toxicity of the crude alkaloids of Toddalia asiatica and to provide pharmacological data for the further development of this herbal medicine. METHODS: We observed the anti inflammatory effects of the crude alkaloids of Toddalia asiatica, using xylol and agra to induce the turgidness and sodium carboxymethyl cellulose (CMC-Na) to induce leucocyte strolling in the rats. The analgesic effects were observed by body-distortion methods. The effects of alkaloids of Toddalia asiatica on hepatic function were observed by testing the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum and calculating the liver index. The LD 50 and 95% creditability were calculated with developed Karber Method. RESULTS: The administration of alkaloids of Toddalia asiatica had the function of inhibiting the auricle swelling caused by xylol and joint swelling caused by agar and leucocyte migration caused by CMC-Na, decreasing the body-distortion of the rats. After two weeks administration, the contents of ALT and AST showed that there was no obvious difference between administered group and control group. The LD50 of the crude alkaloids of Toddalia asiatica was 1.622 g/kg and the 95% creditability was 1.29-2.03 g/kg. CONCLUSION: Toddalia asiatica has anti inflammatory and analgesic effects, and there is no injury to the liver after long-term administration in rats. PMID- 15539028 TI - [Effect of acupuncture serum on intracellular Ca2+ concentration in cultured neurons]. AB - OBJECTIVE: To study the humoral factor of acupuncture in improving functional activities of the organism. METHODS: Neurons of the cerebral cortex from the new born rats were taken to be cultured in a strictly controlled medium. Seven to ten days later, fluorescein-molecular probe Fluo-3AM was used for staining intracellular Ca(2+). Ca(2+) concentration and its changes after application of acupuncture serum were measured by using a laser confocal microscope. Normal serum and acupuncture serum were derived from the normal control rats and the rats undergoing electro-acupuncture of "Baihui (GV 20)", "Zusanli (ST 36)", "Quchi (LI 11)" and "Sanyinjiao (SP 6)" for 2 weeks. RESULTS: After application of normal serum, the intracellular Ca(2+) concentration increased strikingly immediately at the beginning, and gradually got stable afterwards. When acupuncture serum was added to the culture fluid, Ca(2+) concentration lowered significantly. CONCLUSION: These results suggest that bioactive substances in serum collected from rats undergoing acupuncture can affect the intracellular Ca(2+) concentration of the cerebral neurons, and the humoral factor may participate in the effect of acupuncture in regulating functional activities of the organism. PMID- 15539029 TI - [A brief introduction to Shengjiang Power, a traditional Chinese recipe]. PMID- 15539030 TI - [Effect of ultra-fine powder technique on dissolution rates of andrographolide and dehydroandrographolide in Andrographis paniculata]. AB - OBJECTIVE: To investigate the effects of ultra-fine powder technique on dissolution rates of the components in Andrographis paniculata. METHODS: High performance liquid chromatography was employed to determine the concentration of andrographolide and dehydroandrographolide in common powdered or ultra-fine powdered Andrographis paniculata. RESULTS: The dissolution rates of andrographolide and dehydroandrographolide in ultra-fine powdered Andrographis paniculata were higher than those of the general powder. CONCLUSION: Ultra-fine powder technique promotes the dissolution rates of andrographolide and dehydroandrographolide. PMID- 15539031 TI - [Progress in research on chronic fatigue syndrome]. PMID- 15539032 TI - [Progress in research on Ganoderma lucidium spore]. PMID- 15539033 TI - [History and current state of moxibustion]. AB - Moxibustion is an important invention of the Chinese nation, which originated as early as in the clan commune period of the primitive society. The literature records on moxibustion can be traced back to the Warring States Period (475 B.C. to 221 B.C.). Doctors through the ages made considerable progress and published a great number of books on moxibustion. Moxibustion has been applied in treating a great range of diseases. Since the 1950s, the treatment scope of moxibustion has been expanded, and the therapeutic methods of moxibustion are becoming increasingly rich and varied, and great progress in research on the mechanism of moxibustion has been made. Moxibustion was once popular in Europe after its dissemination to the West in the seventeenth century, and the practitioners invented some new methods of moxibustion. Japan is the country in the West where fruitful efforts have been made in research on moxibustion. In modern times, moxibustion has been used for health protection, and the scientists are paying great attention to the experimental research on moxibustion. PMID- 15539034 TI - [Principles for English translation of the terms in traditional Chinese medicine]. PMID- 15539035 TI - [Addition of words in English-Chinese translation for traditional Chinese medicine]. PMID- 15539036 TI - [Discrimination of forms and meanings of the characters in translating ancient books of traditional Chinese medicine into English]. PMID- 15539037 TI - Accuracy of recall of exercise counseling among primary care patients. AB - BACKGROUND: In evaluating the efficacy of health care provider counseling to encourage patients to modify health behaviors such as physical activity, it is important to be able to accurately measure the extent of health care provider counseling. METHODS: The Physical Activity Exit Interview (PAEI) is a brief measure of a patient's perception of the content of physical activity promotion counseling received during a visit with his or her physician. Forty-three primary care patients, and their physicians, completed a PAEI following a visit, which was compared to an audiotape of the visit that was coded to identify the physical activity counseling steps taken. RESULTS: Participants were 67% female, 81.7% white, and had a mean age of 47.1 years. Overall, there was good concordance in the overall number of counseling activities reported between patients and audiotapes (r = 0.47, P < 0.01), patients and physicians (r = 0.51, P < 0.01), and between physicians and audiotapes (r = 0.57, P < 0.01). Significant differences between the three measurement methods (patient exit interview, physician exit interview, audiotape) existed for only 4 of 12 items. CONCLUSIONS: The PAPEI was overall accurate in measuring the content of physical activity counseling, though accuracy differed between items. When discrepancy occurred, it was typically due to patient overreporting of counseling steps. PMID- 15539038 TI - Prevalence of type 2 diabetes mellitus, other cardiovascular risk factors, and cardiovascular disease in Turkish and Moroccan immigrants in North West Europe: a systematic review. AB - BACKGROUND: The prevalence of diabetes, other cardiovascular risk factors, and cardiovascular morbidity and mortality varies between immigrant groups in Western societies, but epidemiological data on these topics are scarce for Turks and Moroccan immigrant living in North West Europe. METHODS: Medline and Embase were systematically searched for studies containing data on the prevalence of diabetes, cardiovascular risk factors, and cardiovascular morbidity and mortality in Turkish or Moroccan immigrants living in Northwestern European countries. RESULTS: Eighteen studies were identified. Corresponding findings were a high prevalence of type 2 diabetes in Turkish and Moroccan immigrants, a high prevalence of smoking among Turkish men, and a very low prevalence of smoking in Moroccan women compared to the indigenous population. Because of lack of valid studies, no definite conclusions could be drawn for in particular blood pressure and lipids. One German study showed exceptionally lower cardiovascular mortality rates in Turkish immigrants. CONCLUSION: The reviewed studies yielded insufficient evidence for a good quality comparison of the cardiovascular risk profile between Turkish and Moroccan immigrants and indigenous populations. Diabetes mellitus was more prevalent in Turkish and Moroccan immigrants, smoking more prevalent in Turkish males, and very rare in Moroccan females. PMID- 15539039 TI - Does knowledge about sexually transmitted infections increase the likelihood of consistent condom use? AB - BACKGROUND: We aimed to assess the association between knowledge about sexually transmitted infections (STIs) and consistent condom use among university students controlling for selected socio-demographic factors. METHOD: An anonymous questionnaire survey was carried out among 720 undergraduate students (76% women) at the University of Tirana, Albania, in October-November 2002. The questionnaire included socio-demographic data, 10 multiple-choice test questions on knowledge about STIs, sexual activity and use of condoms. Two hundred seventy-nine students reported having sexual experience and provided data on condom use as well as knowledge about STIs. Binary logistic regression was used to assess the "independent" association between knowledge about STIs and condom use. RESULTS: In multivariable models, there was an overall association of knowledge about STIs with consistent condom use (OR = 1.74, 95% CI = 1.37-2.23, P < 0.001). There was evidence for an interaction between knowledge and parental education (P = 0.04): there was a positive association among students with highly educated parents (OR = 1.48, 95% CI = 1.11-1.99, P = 0.01), and particularly so among students with low-and-middle educated parents (OR = 4.75, 95% CI = 2.03-11.08, P < 0.01). CONCLUSIONS: Knowledge about STIs is an independent predictor of consistent condom use among university students. Augmentation of knowledge per se should be a component of school-based prevention programs along with the intensification of efforts toward improving students' sexual practices. PMID- 15539040 TI - Influence of demographic, physiologic, and psychosocial variables on adherence to a yearlong moderate-intensity exercise trial in postmenopausal women. AB - BACKGROUND: Few studies have examined the long-term adherence to a yearlong exercise intervention among postmenopausal women. We examined the patterns of adherence to a yearlong exercise intervention and the influence of demographic, physiologic, and psychosocial variables on patterns of adherence among 173 sedentary, overweight, postmenopausal women. METHODS: We collected demographic, physical activity (PA), physiologic, psychosocial, and medical history information at baseline and 12 months. The exercise prescription consisted of at least 45 min of moderate-intensity exercise 5 days/week for 12 months. We calculated several adherence variables. Associations between baseline variables and adherence levels were assessed in bivariate analyses and in multiple regression models. RESULTS: Women randomized to the exercise group (N = 87) participated in moderate-intensity sports or recreational PA on 3.7 +/- 1.4 days/week (79% of the prescribed 5 days/week) for 171 +/- 88 min/week (87% of the prescribed 225 min/week) over the yearlong trial period. Sixty-eight percent of the exercisers had a yearlong average PA level exceeding the national recommendation of 150 min/week. Being in the preparation stage vs. the contemplation stage of the transtheoretical model and a history of participating in any sports or recreational PA were significant predictors of adherence. CONCLUSIONS: Our findings provide important information for the design of future PA interventions and health promotion programs. PMID- 15539041 TI - Smoking abstinence after hospitalization: predictors of success. AB - BACKGROUND: Our objective was to explore the relationship between baseline characteristics of hospitalized smokers and 6-month to 2-year self-reported quit rates. METHODS: We surveyed adult smokers (n = 154) admitted to the Medicine service of an urban public hospital. We used the pharmacy database, a follow-up telephone survey, and medical records to characterize nicotine patch use and post discharge smoking abstinence. RESULTS: Among the 102 patients for whom smoking status at least 6 months after discharge was known, 18 (18%) were not smoking at last contact (mean follow-up 20 months). Individual factors associated with quitting include confidence to quit within 1 week, stage of change other than precontemplation, filling a nicotine patch prescription after discharge, number of previous quit attempts, and increasing age. With multivariate modeling, only confidence to quit [OR 9.8, confidence interval (CI), 2.8-35.0] and the number of previous quit attempts (OR 1.3 per attempt, 95% CI, 1.0-1.5) remained significantly associated with future abstinence. CONCLUSIONS: A high level of confidence to quit and multiple prior quit attempts are strongly associated with future abstinence among hospitalized patients who smoke. Using a simple confidence-to-quit scale to target interventions to patients with high confidence may improve the effectiveness of smoking cessation programs. PMID- 15539042 TI - Television viewing and smoking volume in adolescent smokers: a cross-sectional study. AB - BACKGROUND: Previous research has shown a relationship between television viewing and smoking initiation. The relationship between television viewing and the amount of cigarettes consumed by adolescent smokers per time unit (day, week, month...) has not yet been studied. METHODS: A cross-sectional sample of children by means of self-reports administered by research assistants in schools was obtained. Participants were 421 smokers in a random sample of 4th year students in 15 secondary schools in Flanders, Belgium. Main outcome measures were quantifiable, closed survey questions about smoking volume, whether or not parents and friends smoked, frequency of going out and average weekly TV viewing volume. RESULTS: Television viewing was a significant predictor of smoking volume. Smokers who watch more TV smoke more. The relationship was curvilinear (quadratic). The relationship becomes stronger (curves upward) for higher levels of viewing. Those who watch 5 or more hours a day smoke between 60 and 147 cigarettes more per week than those who watch 1 h or less. CONCLUSIONS: Television viewing is significantly related to smoking volume. The content of television may glamorize smoking. Children may learn to associate smoking with viewing regardless of content. It is also possible that heavier smoking leads to more viewing or that a third variable influences both smoking volume and viewing. Regardless of the causal direction of the relationship television viewing appears to be an indicator or predictor of smoking volume. The curvilinear nature of the relationship deserves further attention. PMID- 15539043 TI - Reported willingness among adolescent nonsmokers to help parents, peers, and others to stop smoking. AB - BACKGROUND: This study of 1025 adolescent nonsmokers aged 11-19 years examined level of interest and factors associated with reported willingness to help someone stop smoking. METHODS: Data were collected from a survey distributed primarily in the schools at four geographic and ethnically diverse study sites. RESULTS: A total of 692 adolescents identified someone close to them who smokes whom they thought should quit. Of these, 90% reported that they would be willing to help this person stop smoking. Multivariate predictors of willingness to help were female gender, less difficulty reading English, and greater level of comfort with talking to the smoker about their smoking. The smoker that the adolescents were willing to help was most often a parent or same age friend. CONCLUSIONS: If this strong interest among adolescents could be tapped, engaging teens as support persons could be a novel public health approach to reaching parents, adolescents, and other smokers in the population. PMID- 15539044 TI - Factors influencing intention to obtain a genetic test for a hereditary disease in an affected group and in the general public. AB - BACKGROUND: To ensure successful implementations of genetic screening in the future, the attitudes of the public are an important factor to consider. The primary aim of this study is to investigate the intention to take a genetic test for an unidentified hereditary disease. A further objective is to assess the predictive values of attitudes, subjective norms, and perceived personal control on the intention to take a genetic test. These aims are investigated in two groups differing in their experience and knowledge of genetic testing. METHOD: A questionnaire was developed according to the Theory of Planned Behavior (TPB) and mailed to a random sample of 1000 persons from the general public and to 330 persons in FAP families. The response rate was 60% and 74%, respectively. RESULTS: The probability of taking a genetic test was high in both groups but significantly higher in the FAP group. The attitudes of the FAP group were significantly more positive when compared to the attitudes of the general public. For the persons in the FAP group, the most significant others in the decision to take a genetic test were their children, whereas spouses proved to be the most important significant others in the general public. The most important predictor of the intention to take a test in both groups was attitude, accounting for 64% of the variance. CONCLUSIONS: The study indicated that most of the individuals in the FAP group and many in the general public intended to take a genetic test. Our findings suggest that living in an affected group and having some kind of experience of a hereditary disease may lead to an even more positive attitude to genetic testing. Using the TPB, attitudes were found to be the strongest predictor of intention to take a genetic test in both groups. PMID- 15539045 TI - Age, gender, and urban-rural differences in the correlates of physical activity. AB - BACKGROUND: The majority of the population is inactive, and strategies to date for promoting regular physical activity have been limited in their effectiveness. Further research is needed to identify correlates of physical activity in different subgroups to design more efficacious interventions. This study sought to identify correlates of physical activity across men and women, urban and rural geographical locations, and four distinct age groups (18-25; 26-45; 46-59; and 60+). METHODS: This study employed data from a large provincial household random sample (N = 20,606) of Canadians. Analyses were utilized to examine the amount of variance explained in self-reported physical activity by a number of demographic and/or biological, psychological, behavioral, social, and environmental variables within each subgroup. RESULTS: Proportion of friends who exercise, injury from past physical activity, educational level, perceived health status, and alcohol consumption were among the strongest correlates across subgroups. CONCLUSIONS: A number of correlates were identified as being significant across all subgroups examined. Most differences in the correlates of physical activity were found within different age groups rather than among urban and rural residents and gender. PMID- 15539046 TI - Cooccurrence of lifestyle risk factors and the explanation of education inequalities in mortality: results from the GLOBE study. AB - BACKGROUND: To investigate whether the cooccurrence of two lifestyle risk factors (smoking, excessive alcohol consumption, physical inactivity in leisure time) has an additional contribution to the explanation of education inequalities in mortality, over and above the contribution of single risk factors. METHODS: Prospective cohort study, 1991-1998, in the South East of the Netherlands. Participants were 16,980 men and women aged 15-74 years at baseline. RESULTS: Education differences in the cooccurrence of risk factors were of a similar magnitude as education inequalities seen for single risk factors. A significant (P = 0.04) interaction effect on mortality was found between smoking and physical inactivity. Adjustment for both smoking and inactivity reduced the mortality hazard ratio of the lowest level of education by 30% (from 1.66 to 1.46). Further adjustment for the interaction between the two risk factors did not change the hazard ratio significantly. CONCLUSION: The cooccurrence of lifestyle risk factors did not provide any additional contribution to the explanation of education inequalities in mortality, over and above that of single risk factors. However, because risk factors tend to cooccur and have a higher prevalence among lower-educated people, it is still useful to focus interventions on more than one risk factor. PMID- 15539047 TI - Relationships between brachial-ankle pulse wave velocity and conventional atherosclerotic risk factors in community-dwelling people. AB - BACKGROUND: A simple instrument has been developed to measure brachial-ankle pulse wave velocity (baPWV). The aim of the present study was to use this instrument to study the relationship between baPWV and conventional atherosclerotic risk factors. METHODS: Community-dwelling Japanese (632) living in a rural area (234 men and 398 women) participated in a municipal medical health survey that included baPWV measurement and a traditional clinical examination, conducted in June, 2002. RESULTS: Men had a significantly higher baPWV than women. No interaction between gender and age on baPWV was identified. Multiple linear regression analysis indicated that age, hemodynamic factors (diastolic blood pressure, pulse pressure, and heart rate), hemoglobinA1c, current drinking and smoking status, and mild retinal changes had significant independent influences on higher baPWV. CONCLUSIONS: In this rural population, age, gender, and hemodynamic factors were independently associated with baPWV, along with traditional atherosclerotic risk factors, although no significant associations between baPWV and histories of atherosclerotic diseases or subclinical atherosclerosis except for mild retinal changes were demonstrated. PMID- 15539048 TI - The use of organizational data for the evaluation of mass prevention programs. AB - BACKGROUND: The effectiveness of health promotion campaigns is hard to measure due to complex outcome and external factors. This study presents a method to evaluate a mass women's health promotion campaign held in a large health maintenance organization (HMO) in Israel. METHODS: This population-based study used administrative and medical databases to examine whether postal invitation to 120,231 HMO-female members increased adherence with certain preventive medicine recommendations (LDL-C, bone density test, and mammography breast cancer screening). A comparison was made using three different reference data: pre- and post-campaign periods (1998-2003), HMO-male members who were not targeted by the campaign, and rates of urine tests, which were also not targeted by the campaign. RESULTS: During the 2 months following the campaign, adherence with mammography (3.8%) and LDL-C (12.5%) reached their maximum rates in 5 years. Adherence with bone density test increased from 2.3% in 2000 to 2.8% in the campaign period. No similar trends were observed for urine or LDL-C tests among men. CONCLUSIONS: The use of multiple reference groups through the analysis of administrative and medical databases supports the association between the campaign and improved adherence with screening tests. A similar methodology may be adopted for the analysis of mass health promotion campaigns in large HMOs. PMID- 15539049 TI - Effects of combined sensory and muscular training on balance in Japanese older adults. AB - BACKGROUND: Adequate levels of physical balance and muscular strength are necessary to live independently in old age. The effects of an exercise training program targeting the sensory and muscle systems on balance and strength in a group of older adults were determined in this study. METHODS: Static balance (one leg balance with eyes closed), dynamic balance (limits of stability [endpoint excursion [EPE], maximum excursion [MXE]]), and strength (chair stand) were assessed before and after the intervention. Volunteers were divided randomly into a training group (TR, n = 15, 76 +/- 4 years) and a control group (CN, n = 14, 76 +/- 7 years). TR performed 12-week (2 days/week, 60 min/day) supervised training while standing on the floor (first 4 weeks) and progressing to standing on foam pads of different compliances. Exercises included movements that challenged the sensory and muscle systems. RESULTS: After 12 weeks of training, the TR demonstrated significant improvements in static balance (82%); EPE backward (72%), right (32%), and left (33%); MXE backward (74%), right (31%), and left (18%); and lower body muscle strength (20%) with no significant changes in CN. CONCLUSION: These results indicate that this training program is very effective in improving balance and lower body strength in older adults. PMID- 15539050 TI - Late relapse/sustained abstinence among former smokers: a longitudinal study. AB - BACKGROUND: Relapse remains the most refractory aspect of nicotine dependence, but little is known about relapse that occurs after a period of prolonged abstinence (i.e., "late relapse"). METHOD: The current study examined the prevalence and predictors of late relapse/sustained abstinence in a community based, longitudinal cohort of former smokers surveyed at baseline and again 4 years later (n = 1143). RESULTS: Late relapse rates declined precipitously and sustained abstinence increased as the duration of abstinence at baseline increased. Although the risk for late relapse appeared to be small when using a point prevalence definition of smoking, that risk increased substantially if fluctuations in smoking behavior over time were considered (i.e., many former smokers relapsed and then regained abstinence during the 4-year study period). Although there were a number of significant prospective predictors of late relapse/sustained abstinence, they did not retain their predictive ability after controlling for baseline duration of abstinence. CONCLUSION: Although late relapse rates in the general population of former smokers appear low, and particularly so for individuals who have been abstinent for long periods of time, fluctuations between abstinence and smoking over time are not uncommon. How these fluctuations influence health or sustained abstinence over time is unclear. Because late relapse rates are generally low, interventions are likely to require careful targeting of at-risk individuals if they are to be efficacious and cost effective. PMID- 15539051 TI - Health values and health-information-seeking in relation to positive change of health practice among middle-aged urban men. AB - BACKGROUND: Preventive health practice is an important component of health promotion strategy. The objectives of this cohort study were to assess changes of health practices over 3 years among middle-aged Japanese men and to examine the factors related to their positive changes in lifestyle. METHODS: Three hundred thirty-four randomly selected men in Tokyo completed two face-to-face structured interviews, one in 1998 and the other in 2001. Daily health practices, health values, health-information-seeking, socioeconomic characteristics, and health status were assessed. A Health Practice Index (HPI) was calculated to reflect overall health behavior. According to the change of HPI, the mode of change in general health practice between the first and second interview was classified as positive change, no change, and negative change. RESULTS: Percentages of the subjects who engaged in eight identified healthy practices ranged from 31.7% to 54.5% at baseline. The follow-up interval revealed the adoption of each particular healthy or unhealthy practice as 5.7% to 33.6%. A high value placed on health independently associated with positive change of general health practice (OR = 2.95, 95% CI = 1.23 to 7.08), and inversely associated with negative change (OR = 0.45, 95% CI = 0.18 to 1.10); consciously seeking health information associated with positive change (OR = 2.16, 95% CI = 1.07 to 4.36) after controlling for socioeconomic and health status. CONCLUSION: Considerable opportunity remains for urban middle-aged men to further improve their health behavior. Health values saliency, sensitively designed health information, and health status perception, as well as socioeconomic status, should be considered for successful promotion of healthy lifestyle among the adult male population. PMID- 15539052 TI - Induction of apoptosis by epigallocatechin-3-gallate via mitochondrial signal transduction pathway. AB - PURPOSE: To elucidate the apoptosis induction of Epigallocatechin-3-gallate (EGCG) on nasopharyngeal carcinoma (NPC) cells via mitochondrial signal transduction pathway regulated by EB-virus-encoded latent membrane protein 1 (LMP1). METHODS: The survival rates of pTet-on-LMP1 HNE2 cells after the EGCG treatment were determined by MTT assay. Induction of apoptosis in pTet-on-LMP1 HNE2 cells after the EGCG treatment was analyzed by agarose gel electrophoresis. The activity of caspase-9 was determined by ApoAlert Caspase-9 Fluorescent Assay kit after the EGCG treatment. The protein expressions of cytochrome c and Bcl-2 were analyzed by Western blotting after the EGCG treatment. RESULTS: EGCG inhibited the survival rates of pTet-on-LMP1 HNE2 cells and induced apoptosis of pTet-on-LMP1 HNE2 cells. EGCG raised the activities of caspase-9, enhanced the releasing of cytochrome c from the mitochondria, and suppressed the protein expression of Bcl-2. These are the key targets on the mitochondrial signal transduction pathway in apoptosis. CONCLUSIONS: EGCG inhibited the survival rate of NPC cells and induced apoptosis of NPC cells via the mitochondrial signal transduction pathway. This study suggests that the interference effect of EGCG on targets of the mitochondrial signal transduction pathway plays an important role in the anticancer function. PMID- 15539053 TI - Quit attempts among African American teenage smokers seeking treatment: gender differences. AB - BACKGROUND: African Americans experience disproportionate smoking-related mortality. Because established smoking during youth predisposes to adult smoking and serious health consequences, characterizing ethnic differences in adolescent smokers' self-quit attempts may inform ethnic-specific approaches to youth smoking cessation. METHODS: African American and European American teenage smokers applying to a teenage smoking cessation study (2000-2003) provided smoking-related data, including characteristics of previous cessation attempts and prior use of nicotine replacement therapy (NRT). Tobacco dependence was assessed using the Fagerstrom Test of Nicotine Dependence (FTND). RESULTS: Of 980 (15.5 +/- 1.3 years, 41.8% African American, 59.9% female) participants, African Americans boys were significantly less likely than European American boys to report a prior quit attempt (OR = 0.35, 95% CI 0.17-0.73, P = 0.0049) or to have used NRT (OR = 0.60, 95% CI 0.36-0.998, P = 0.049) after adjusting for years smoked and FTND score. African American girls were more likely to report a prior request for cessation treatment than European American girls after adjusting for FTND and years smoked (OR = 2.19, 95% CI 1.37-3.48, P = 0.001). CONCLUSIONS: While increasing education and outreach to African American boys and enhancing access to formal cessation programs for African American girls who smoke may be beneficial, our findings warrant extension to non-treatment-seeking teenage smokers. PMID- 15539054 TI - Factors influencing the wearing of facemasks to prevent the severe acute respiratory syndrome among adult Chinese in Hong Kong. AB - BACKGROUND: The global outbreak of the severe acute respiratory syndrome (SARS) in 2003 has been an international public health threat. Quick diagnostic tests and specific treatments for SARS are not yet available; thus, prevention is of paramount importance to contain its global spread. This study aimed to determine factors associating with individuals' practice of the target SARS preventive behavior (facemask wearing). METHODS: A total of 1329 adult Chinese residing in Hong Kong were surveyed. The survey instrument included demographic data, measures on the five components of the Health Belief Model, and the practice of the target SARS preventive behavior. Logistic regression analyses were conducted to determine rates and predictors of facemask wearing. RESULTS: Overall, 61.2% of the respondents reported consistent use of facemasks to prevent SARS. Women, the 50-59 age group, and married respondents were more likely to wear facemasks. Three of the five components of the Health Belief Model, namely, perceived susceptibility, cues to action, and perceived benefits, were significant predictors of facemask-wearing even after considering effects of demographic characteristics. CONCLUSIONS: The Health Belief Model is useful in identifying determinants of facemask wearing. Findings have significant implications in enhancing the effectiveness of SARS prevention programs. PMID- 15539055 TI - Joint impact of health risks on health care charges: 7-year follow-up of National Health Insurance beneficiaries in Japan (the Ohsaki Study). AB - BACKGROUND: The objective of this study was to examine the joint impact of modifiable health-risk factors such as smoking, obesity, and physical inactivity on direct health care charges. METHOD: We conducted a population-based prospective cohort study, with follow-up from 1995 to 2001. The participants were Japanese National Health Insurance (NHI) beneficiaries (26,110 men and women aged 40-79 years). RESULTS: 'No risk' group defined as never-smoking, body mass index (BMI) 20.0-24.9 kg/m(2), and walking for >/=1 h/day had mean health care charges of 171.6 dollars after adjustment for potential confounders. Compared with this group, the presence of smoking (SM; ever-smoking) alone, obesity alone (OB; BMI >/=25.0 kg/m(2)), or physical inactivity (PI; walking for <1 h/day) alone were associated with a 8.3%, 7.1%, or 8.0% increase in health care charges, respectively. The combinations of the risks of SM and OB, SM and PI, OB and PI, and SM and OB and PI were associated with a 11.7%, 31.4%, 16.4%, and 42.6% increase in charges, respectively. CONCLUSION: Interventions to improve modifiable health-risk factors may be a cost-effective approach for reducing health care charges as well as improving people's health. PMID- 15539056 TI - The association between physical exercises and health-related quality of life in subjects with mental disorders: results from a cross-sectional survey. AB - OBJECTIVES: The present study examines the relationship between health-related quality of life and physical activity among adults with affective, anxiety, and substance dependence disorders. METHODS: Analyses were conducted among participants in the German National Health Interview and Examination Survey (GHS), a nationally representative multistage probability survey, conducted from 1997 to 1999. Multiple linear regression analyses were used to determine the relationship between health-related quality of life and physical activity among subjects with mental disorders. RESULTS: Affective, anxiety, and substance dependence disorders were associated with substantial impairment in health related quality of life. Higher levels of physical activity were associated with higher health-related quality of life among persons with mental disorders. Even after controlling for sociodemographic characteristics, physically inactive subjects reported poorer quality of life. CONCLUSIONS: Physical activity can be considered as beneficial for people suffering from mental disorders. The promotion of a physically active lifestyle is an important public health objective. PMID- 15539057 TI - Behavioral interventions and cost-effectiveness analysis. AB - Behavioral health interventions are often gauged with a dichotomous outcome, "success" or "failure." Hidden by this dichotomy is a series of behavior changes that can be followed with the Transtheoretical Model (stages of change). There has been little consideration, however, about whether this information can and should be used in cost-effectiveness analysis. We review the stages of change model and its applications to behavioral health interventions. We then discuss analytical methods for including stages of change, or similar behavior change models, in cost-effectiveness analysis (CEA). This is typically not done but it may be critical for study design and for interpreting CEA results. PMID- 15539058 TI - Health benefits of a pedometer-based physical activity intervention in sedentary workers. AB - BACKGROUND: Inactivity is a leading contributor to chronic health problems. Here, we examined the effects of a pedometer-based physical activity intervention (Prince Edward Island-First Step Program, PEI-FSP) on activity and specific health indices in 106 sedentary workers. METHODS: Participants were recruited from five workplaces where most jobs were moderately-highly sedentary. Using subjects as their own control, physical activity (pedometer-determined steps per day) was compared before and after a 12-week intervention. Changes in body mass index (BMI), waist girth, resting heart rate, and blood pressure were evaluated. RESULTS: The PEI-FSP was completed by 59% of participants. Steps per day increased from 7,029 +/- 3,100 (SD) at baseline to a plateau of 10,480 +/- 3,224 steps/day by 3.96 +/- 3.28 weeks of the intervention. The amount that participants were able to increase their steps per day was not related to their baseline BMI. On average, participants experienced significant decreases in BMI, waist girth, and resting heart rate. Reductions in waist girth and heart rate were significantly related to the increase in steps per day. In contrast, reductions in BMI were predicted by the initial steps per day. CONCLUSIONS: The PEI-FSP increased physical activity in a sedentary population. Importantly, those with a higher BMI at baseline achieved relatively similar increases in their physical activity as participants with a lower BMI. PMID- 15539059 TI - Declining seroprevalence of rubella antibodies among young Israeli adults: a 12 year comparison. AB - BACKGROUND: In March 2000, the Israel Defense Force (IDF) experienced an outbreak of rubella. No population-based rubella seroepidemiologic data were available for the 13-year period preceding the epidemic. METHODS: We conducted a population based seroprevalence study of rubella antibodies among 289 IDF recruits drafted in 1999. RESULTS: We found that 69.2% of males and 90.8% of females in service were seropositive at recruitment in 1999, immediately before the outbreak. These rates were significantly lower than those last measured in 1987 (88% among males and 98.1% among females). CONCLUSIONS: Our findings indicate that this rubella outbreak among young adults was facilitated by a decrease in immunity to levels below those required for herd immunity, at which epidemic virus transmission was no longer blocked. This is most likely due to widespread pediatric vaccination coverage with incomplete catch-up immunization among adolescents and young adults. These findings serve as a reminder that changes to childhood vaccination programs may affect the epidemiology of disease among older segments of the population several years after the change is implemented. PMID- 15539060 TI - Factors associated with condom use among young Denver inner city women. AB - BACKGROUND: Despite the availability of condoms and theoretically based interventions to promote their use, sexually active women aged 15 to 25 years continue to put themselves at risk for sexually transmitted diseases and unintended pregnancy. METHODS: One hundred ninety-eight inner city women were interviewed about knowledge and attitudes about condoms. Using the Transtheoretical Model, regression techniques were used to identify factors associated with condom use at last sex and the proportion of acts protected by a condom in the last 90 days. RESULTS: Constructs including intention to use (OR = 1.69, CI 1.07-2.65) and positive outcome expectancies (OR = 1.59, CI 1.03-2.46) were associated with condom use at last act of sexual intercourse. Similarly, intention to use condoms (RR = 1.58, CI 1.37-1.82), positive outcome expectancies (RR = 2.71, CI 2.41-2.99), perceived peer's use of condoms (RR = 2.25, CI 1.95 2.60), and number of places condoms were discussed (RR 1.05, CI 1.02-1.07) were associated with the proportion of protected acts. CONCLUSIONS: Constructs specified in the Transtheoretical Model are useful in describing condom use and have implications for targeting human immunodeficiency virus (HIV)/sexually transmitted diseases (STD)/unintended pregnancy interventions. PMID- 15539061 TI - Healthy skepticism. AB - OBJECTIVE: To investigate the relationship between medical skepticism and overall self-rated health and to identify disparities in health for vulnerable subgroups among the elderly. DESIGN: A cross-sectional telephone survey involving multiple callbacks. Independent variables included three measures of medical skepticism and disparities variables (low income, low education, race/ethnicity, gender, rural residence) along with several control variables (body weight, marital status, employment, insurance coverage, number of medical visits). SETTING: West Texas, a sparsely populated 108-county region. PARTICIPANTS: Five thousand six persons aged 65 and over. MAIN RESULTS: Multiple logistic regression analysis revealed that medical skepticism (believing that one can overcome illnesses without the help of a medical professional) was independently related to better self-rated overall health. Disparities in health were found for income, race/ethnicity, and low education but not for residents of rural or frontier areas (vs. urban residents). CONCLUSIONS: Belief in one's own ability to manage most illnesses may or may not be causally related to better health. However, the association is promising and deserves further investigation. Programs promoting self-care among groups facing health disparities should be considered. PMID- 15539062 TI - Trends in cigarette smoking among US adults with diabetes: findings from the Behavioral Risk Factor Surveillance System. AB - BACKGROUND: Smoking substantially increases morbidity and mortality rates in people with diabetes. Previous studies have shown that the prevalence of smoking among people with diabetes is similar to that among people without diabetes. We sought to examine temporal trends in the prevalence of smoking among people with diabetes since 1990. METHODS: We analyzed data from the Behavioral Risk Factor Surveillance System for 1990-2001. RESULTS: The age-adjusted prevalence of smoking among adults with diabetes was 23.6% (men, 25.4%; women, 22.2%) in 1990 and 23.2% (men, 24.8%; women, 21.9%) in 2001. In comparison, the prevalence among participants without diabetes was 24.2% (men, 25.7%; women, 22.8%) in 1990 and 23.2% (men, 24.8%; women, 21.5%) in 2001. Thus, the prevalence of cigarette smoking was similar and remained stable from 1990 through 2001. Among participants with diabetes, significant decreases in the prevalence of smoking occurred among African Americans and those aged >/=65 years. CONCLUSIONS: New efforts and commitments to promote smoking cessation among people with diabetes are needed. PMID- 15539063 TI - Prescription weight loss pill use among Americans: patterns of pill use and lessons learned from the fen-phen market withdrawal. AB - BACKGROUND: Despite the popularity of antiobesity medications, there is a lack of population-based data on their use. In addition, response (termination of pill use and receipt of an echocardiogram) to the fenfluramine and dexfenfluramine market withdrawal among the public has not been described. Lessons learned from this event have implications for future withdrawals. METHODS: We used data from the Behavioral Risk Factor Surveillance System (BRFSS) a random-digit telephone survey. In 1998, six states included detailed questions about the use of prescription weight loss pills in the previous 2 years, n = 16,460 noninstitutionalized adults aged 18 years or older. RESULTS: Almost one third of prescription weight loss pills users were not obese before taking pills. Family and friends and other nonphysicians were reported as sources of medication by one in ten users. One third of users also reported taking nonprescription diet products. Among fenfluramine or dexfenfluramine users, one third continued pill use after the market withdrawal and only one quarter received echocardiograms. CONCLUSIONS: Despite enormous publicity, many persons continued to use fen-phen after the market withdrawal and most did not receive follow-up echocardiograms. Our study raises issues regarding the effectiveness of withdrawal warnings in a small but significant subset. Additional means of communicating risk to individuals are needed for future product withdrawals including special strategies for those lacking healthcare coverage. PMID- 15539064 TI - Race, ethnicity, and sociocultural characteristics predict noncompliance with lipid-lowering medications. AB - BACKGROUND: Sociocultural factors that predict noncompliance with lipid-lowering medications are not well understood. METHODS: Interviews and medical record review were conducted for 510 patients treated with lipid-lowering medications. Compliance with lipid-lowering medications was measured by patients' self assessment of medication-taking practices. RESULTS: Adjusted for age and sex, there was a higher frequency of noncompliance among Black and Hispanic subjects. Adjusted for age, sex, and race/ethnicity, noncompliance was associated with medication side effects, feelings of sadness or depression, fair or poor health status, primary use of a language other than English, single or divorced status, fewer and less frequent contact with friends, children in the household, and lower education. Independent predictors of noncompliance in multivariate models were side effects (OR = 3.9, P < 0.01), sadness or depression (OR = 1.9, P = 0.05), Black (OR = 3.7, P < 0.01, vs. White), Hispanic (OR = 6.3, P < 0.01, vs. White), single or divorced (OR = 2.1, P < 0.01), children in the household (OR = 1.5 per child, P < 0.01), and lack of health insurance (OR = 2.4, P = 0.05). CONCLUSIONS: Unmarried status, feelings of sadness or depression, lack of insurance, and children in the household were independently associated with poorer compliance with lipid-lowering medications in this urban population. Nonetheless, higher noncompliance among minority patients persisted independently of these and other specific social, cultural, and economic factors. PMID- 15539065 TI - Cobalamin: a critical vitamin in the elderly. AB - Vitamin B(12) deficiency is a common problem in elderly subjects. If a serum cobalamin level of about 150 pmol/L (200 pg/mL) is considered normal, 10-15% of the elderly are deficient. Today, however, a threshold of 220-258 pmol/L (300-350 pg/mL) is recognized as desirable in the elderly, or else sensitive markers like the blood concentration of homocysteine or methylmalonic acid (MMA) are used. Then the prevalence of cobalamin deficiency rises to up to 43%. In the elderly, this high prevalence of poor cobalamin status is predominantly caused by atrophic gastritis type B. Atrophic gastritis results in declining gastric acid and pepsinogen secretion, and hence decreasing intestinal absorption of the cobalamin protein complexes from food. About 20-50% of the elderly are affected. Furthermore, the reduced acid secretion leads to an alkalinization of the small intestine, which may result in bacterial overgrowth and thus to a further decrease of the bioavailability of the vitamin. In addition, some drugs such as proton pump inhibitors or H2 receptor antagonists inhibit the intestinal absorption of vitamin B(12). An already moderately reduced vitamin B(12) level is associated with vascular disease and neurocognitive disorders such as depression and impaired cognitive performance. Furthermore, a poor vitamin B(12) status is assumed to be involved in the development and progression of dementia (e.g., Alzheimer's dementia). This is especially observable if the folic acid status is reduced as well. Due to the insecure supply, the cobalamin status of elderly persons (>/=60 years) should be regularly controlled and a general supplementation with vitamin B(12) (>50 microg/day) should be considered. PMID- 15539066 TI - [Study and evaluation of patients with obliterating arteriopathy of the lower limbs: use of isokinetics to analyze muscular strength and fatigue]. AB - OBJECTIVES: The aim of the study was to evaluate strength of the dorsal and plantar flexors in patients with obliterating arteriopathy of the lower limbs (stage 2) and to evaluate fatigue. POPULATION AND METHODS: The comparison was based on two groups of ten patients: one with obliterating arteriopathy, and the second with healthy limbs as a control group. Each group of patients underwent bilateral plantar flexion and dorsiflexion of the ankle on an isokinetic machine, with the following movements: three series of five movements at 120 degrees/s, then three series of three movements at 30 degrees/s in a concentric mode to evaluate strength. The exercise was repeated 20 times at 180 degrees/s to evaluate fatigue. The isokinetic parameters selected were the couple peak and ratio of workload to total weight of the patient. Tests controlled for cardiac frequency, as measured by electrocardiography, and blood pressure. RESULTS: Patients with obliterating arteriopathy of the lower limbs were generally weaker in global strength of plantar and dorsal flexors and had greater muscular fatigue than the control group (P < 0.05). CONCLUSION: Decreased performance in the leg muscles in patients with obliterating arteriopathy of the lower limbs is likely due to arterial deficit. Muscular metabolism anomalies such dysfunction of the terminal stages of the oxidative phosphorylation could also affect strength. Use of isokinetics represents an interesting possibility for treatment of patients with multiple cardiovascular abnormalities. PMID- 15539067 TI - [Effects of training on static and dynamic balance in elderly subjects who have had a fall or not]. AB - OBJECTIVE: To evaluate the effects of a physical training program on static and dynamic balance during single and dual task conditions in elderly subjects who have had a fall or not. PATIENTS: Two groups, comprising a total of 33 elderly subjects, were trained: 16 who had a fall were 69.2 +/- 5.0 years old and 17 who had not had a fall were 67.3 +/- 3.8 years. METHODS: All subjects underwent an unipedal test with eyes open and eyes closed, followed by gait assessment during single and dual motor task conditions, before and after a physical training program. RESULTS: All subjects showed a significant decrease, by six times for subjects who had fallen and four times by those who had not, in the number of touch-downs in the unipedal test with eyes open (P < 0.05), and by 2.5 and 2 times, respectively, with eyes closed (P < 0.05) after the training program. All subjects showed a significant increase in speed (P < 0.05), cadence (P < 0.05) and stride length (P < 0.05) and a significant decrease in the single support time (P < 0.05) and stride time (P < 0.05) in gait assessment during single and dual task conditions after the training program. During the training program, no subjects fell. CONCLUSION: The physical training program improved static balance and quality of gait in elderly subjects who had had a fall and those who had not, which could contribute to minimizing and/or retarding the effects of aging and maintaining physical independence. PMID- 15539068 TI - [Temporal, kinetic and kinematic asymmetry in gait initiation in one subject with hemiplegia]. AB - OBJECTIVE: To investigate the temporal, kinetic and kinematic asymmetry of gait initiation in one subject with hemiplegia with an equinus varus foot. MATERIAL AND METHODS: A kinetic analysis with two AMTI force plates and a kinematic analysis with an ELITE optoelectronic system of gait initiation were performed in one subject with hemiplegia. RESULTS: The duration of the gait initiation phases was asymmetrical. The monopodal phase was shorter when the affected lower limb was supporting than when the healthy one was supporting. The propulsion resulted from the force exerted on the healthy lower limb. The distribution of body weight on the lower limbs was asymmetrical. Body weight support was more important on the healthy side than on the affected side. Maximal extension of the ankle on the hemiplegic side occurred during the swing phase. Ground clearance was increased by elevating the knee higher on the affected side than on the healthy side during the swing phase. Initial contact with the floor was performed with the foot flat on the affected side. CONCLUSION: This preliminary study has shown that gait initiation in one subject with hemiplegia was asymmetrical in kinetics and kinematics. The results concerning kinematics have not been reported previously for gait initiation in subjects with hemiplegia. The study of gait initiation should allow for better understanding postural and movement control strategies developed by patients with hemiplegia. PMID- 15539069 TI - [Intradiscal injection of acetate of prednisolone in severe low back pain: complications and patients' assessment of effectiveness]. AB - OBJECTIVES: To investigate complications (calcification and/or disc collapse) as seen on radiography and patient-assessed effectiveness after intradiscal injections of acetate of prednisolone (Hydrocortancyl). MATERIALS AND METHODS: A retrospective study of medical records of 67 patients (68 injected discs) given an intradiscal injection of acetate of prednisolone for low back pain with endplate signal changes on magnetic resonance imaging and/or evidence of fast destructive discopathy, as well as records of 85 patients (55 women, mean age 49 +/-9 years) to assess the effectiveness of intradiscal injection: the global appreciation of the patient (excellent, good, mild, none, worse) concerning the result of the intradiscal injection at one, three and six months. Two physicians were blinded during analysis of images. RESULTS: The mean period of follow up for radiographics was 10.25 +/-5.99 months. A total of 44.8% of the patients had control at six months and 38.8% at 12 months or more. No calcification was found, but two discs showed a collapse of 20 and 25% at six and 12 months, respectively, after the injection. For effectiveness of intradiscal injection, 71.8% the patients considered the result good or excellent at one month, 55.3% at three months and 43.5 % at six months. CONCLUSION: The benefit-to-risk ratio of use of acetate of prednisolone intradiscal injection seems good in selected patients with low back pain. PMID- 15539070 TI - [Impact of frailty factors in elderly people older than 75 years with prosthesis of the hip]. AB - OBJECTIVE: Hip pathology of hip requiring a surgical assumption of responsibility (coxarthrose invalidating, hip fracture) is increasing, and after the year 2025, the number of interventions will double. In parallel, the number of elderly people (older than 75 years) with malnutrition associated with cognitive impairment and reduced autonomy is on the increase. So the concept of frail elderly identified by specific criteria allows for better defining the health needs of heterogeneous elderly people. The objective of this study was to determine the influence of frailty factors on autonomy and clinical outcome after intervention for prosthesis of the hip. METHODOLOGY: This prospective work, completed over 8 months, included 55 patients with total or intermediate prostheses of hip, whatever the cause, recruited from rehabilitation wards. Results from a pre-established questionnaire defined the personal status and social framework of these patients. A medical examination evaluated cognitive function Mini Mental State (MMS), nutritional state (IMC) and the level of autonomy immediately after surgery (the first 2 weeks) by the measurement of functional independence (MIF). The second MIF was carried out by the same operator at the end of the assumption of responsibility by the institution. RESULTS: Three factors of frailty influenced progress at the time of reacquisition of autonomy: cognitive impairment (P < 0001; 1 patient out of 2 [50%] had an MMS score < 22); reduced nutrition? (P < 0.0026; [29% had a BMI score < 20 or a perimeter of the calf < 30 cm]; and accommodations before the intervention (P < 0.028; [15% lived in an institution]). CONCLUSION: Nutritional state, cognitive level and social activity must be evaluated regularly in elderly subjects with hip prosthesis to evaluate optimal load and adaptation. PMID- 15539071 TI - [Effects of frailness on the outcome of people more than 75 years undergoing a hip replacement]. PMID- 15539072 TI - Molecular basis of bacterial resistance to chloramphenicol and florfenicol. AB - Chloramphenicol (Cm) and its fluorinated derivative florfenicol (Ff) represent highly potent inhibitors of bacterial protein biosynthesis. As a consequence of the use of Cm in human and veterinary medicine, bacterial pathogens of various species and genera have developed and/or acquired Cm resistance. Ff is solely used in veterinary medicine and has been introduced into clinical use in the mid 1990s. Of the Cm resistance genes known to date, only a small number also mediates resistance to Ff. In this review, we present an overview of the different mechanisms responsible for resistance to Cm and Ff with particular focus on the two different types of chloramphenicol acetyltransferases (CATs), specific exporters and multidrug transporters. Phylogenetic trees of the different CAT proteins and exporter proteins were constructed on the basis of a multisequence alignment. Moreover, information is provided on the mobile genetic elements carrying Cm or Cm/Ff resistance genes to provide a basis for the understanding of the distribution and the spread of Cm resistance--even in the absence of a selective pressure imposed by the use of Cm or Ff. PMID- 15539073 TI - Lipopolysaccharides of anaerobic beer spoilage bacteria of the genus Pectinatus- lipopolysaccharides of a Gram-positive genus. AB - Bacteria of the genus Pectinatus emerged during the seventies as contaminants and spoilage organisms in packaged beer. This genus comprises two species, Pectinatus cerevisiiphilus and Pectinatus frisingensis; both are strict anaerobes. On the basis of genomic properties the genus is placed among low GC Gram-positive bacteria (phylum Firmicutes, class Clostridia, order Clostridiales, family Acidaminococcaceae). Despite this assignment, Pectinatus bacteria possess an outer membrane and lipopolysaccharide (LPS) typical of Gram-negative bacteria. The present review compiles the structural and compositional studies performed on Pectinatus LPS. These lipopolysaccharides exhibit extensive heterogeneity, i.e. several macromolecularly and structurally distinct LPS molecules are produced by each strain. Whereas heterogeneity is a common property in lipopolysaccharides, Pectinatus LPS have been shown to contain exceptional carbohydrate structures, consisting of a fairly conserved core region that carries a large non-repetitive saccharide that probably replaces the O-specific chain. Such structures represent a novel architectural principle of the LPS molecule. PMID- 15539074 TI - Recent trends on the molecular biology of pneumococcal capsules, lytic enzymes, and bacteriophage. AB - Streptococcus pneumoniae has re-emerged as a major cause of morbidity and mortality throughout the world and its continuous increase in antimicrobial resistance is rapidly becoming a leading cause of concern for public health. This review is focussed on the analysis of recent insights on the study of capsular polysaccharide biosynthesis, and cell wall (murein) hydrolases, two fundamental pneumococcal virulence factors. Besides, we have also re-evaluated the molecular biology of the pneumococcal phage, their possible role in pathogenicity and in the shaping of natural populations of S. pneumoniae. Precise knowledge of the topics reviewed here should facilitate the rationale to move towards the design of alternative ways to combat pneumococcal disease. PMID- 15539075 TI - Non-homologous end-joining factors of Saccharomyces cerevisiae. AB - DNA double-strand breaks (DSB) are considered to be a severe form of DNA damage, because if left unrepaired, they can cause a cell death and, if misrepaired, they can lead to genomic instability and, ultimately, the development of cancer in multicellular organisms. The budding yeast Saccharomyces cerevisiae repairs DSB primarily by homologous recombination (HR), despite the presence of the KU70, KU80, DNA ligase IV and XRCC4 homologues, essential factors of the mammalian non homologous end-joining (NHEJ) machinery. S. cerevisiae, however, lacks clear DNA PKcs and ARTEMIS homologues, two important additional components of mammalian NHEJ. On the other hand, S. cerevisiae is endowed with a regulatory NHEJ component, Nej1, which has not yet been found in other organisms. Furthermore, there is evidence in budding yeast for a requirement for the Mre11/Rad50/Xrs2 complex for NHEJ, which does not appear to be the case either in Schizosaccharomyces pombe or in mammals. Here, we comprehensively describe the functions of all the S. cerevisiae NHEJ components identified so far and present current knowledge about the NHEJ process in this organism. In addition, this review depicts S. cerevisiae as a powerful model system for investigating the utilization of either NHEJ or HR in DSB repair. PMID- 15539076 TI - Biogenesis of peroxisomes and glycosomes: trypanosomatid glycosome assembly is a promising new drug target. AB - In trypanosomatids (Trypanosoma and Leishmania), protozoa responsible for serious diseases of mankind in tropical and subtropical countries, core carbohydrate metabolism including glycolysis is compartmentalized in peculiar peroxisomes called glycosomes. Proper biogenesis of these organelles and the correct sequestering of glycolytic enzymes are essential to these parasites. Biogenesis of glycosomes in trypanosomatids and that of peroxisomes in other eukaryotes, including the human host, occur via homologous processes involving proteins called peroxins, which exert their function through multiple, transient interactions with each other. Decreased expression of peroxins leads to death of trypanosomes. Peroxins show only a low level of sequence conservation. Therefore, it seems feasible to design compounds that will prevent interactions of proteins involved in biogenesis of trypanosomatid glycosomes without interfering with peroxisome formation in the human host cells. Such compounds would be suitable as lead drugs against trypanosomatid-borne diseases. PMID- 15539077 TI - Lipoproteins of Mycobacterium tuberculosis: an abundant and functionally diverse class of cell envelope components. AB - Mycobacterium tuberculosis remains the predominant bacterial scourge of mankind. Understanding of its biology and pathogenicity has been greatly advanced by the determination of whole genome sequences for this organism. Bacterial lipoproteins are a functionally diverse class of membrane-anchored proteins. The signal peptides of these proteins direct their export and post-translational lipid modification. These signal peptides are amenable to bioinformatic analysis, allowing the lipoproteins encoded in whole genomes to be catalogued. This review applies bioinformatic methods to the identification and functional characterisation of the lipoproteins encoded in the M. tuberculosis genomes. Ninety nine putative lipoproteins were identified and so this family of proteins represents ca. 2.5% of the M. tuberculosis predicted proteome. Thus, lipoproteins represent an important class of cell envelope proteins that may contribute to the virulence of this major pathogen. PMID- 15539078 TI - In vitro IL-2 incubation induces CD69 expression and other phenotypic changes on NK subpopulations present in PBPC collections. PMID- 15539079 TI - Senescence and functional failure in hematopoietic stem cells. AB - Maintaining normal function of hematopoietic stem cells (HSCs) is critical to blood coagulation, oxygen transportation, and host defense against infection. A potential cause of HSC dysfunction is senescence, in which HSCs and progenitor cells suffer from proliferative arrest. Studies on humans and various animal models have indicated that HSCs can become senescent, showing a significant decline in functional ability with increasing age. There are genetic elements mapped to specific chromosomal sites that regulate HSC senescence. These elements may differ among species, strains, and even individuals. HSC senescence and related pathological effects may not be obvious during normal lifetime under most circumstances since individual primitive HSC clones can function long-term to produce progeny that sustain life-long mature blood cell production. Shortening of telomeres at the chromosomal ends could contribute to HSC senescence, especially when HSCs are stressed under certain pathological conditions. Future studies should define the molecular elements that are important in the regulation of HSC senescence. PMID- 15539080 TI - Long-term persistent lymphopenia in hematopoietic stem cell donors after donation for donor lymphocyte infusion. AB - OBJECTIVES: To analyze the consequences of lymphocyte donations on lymphopoiesis in donors having previously undergone hematopoietic stem cell collection for allogeneic stem cell transplantation. METHODS: Repeated analysis of leukocyte subsets in the peripheral blood of 76 hematopoietic stem cell donors undergoing lymphocyte donation(s) for DLI. RESULTS: Grade I/II lymphopenia was present in 22 donors (29%) just before first apheresis for lymphocyte collection, demonstrating that former stem cell donation induced prolonged lymphopenia in a subset of donors. The monocytic lineage was not affected. Older age and history of PBSC harvest constituted 2 independent factors of lymphopenia, but had no influence on monocytopenia. The first apheresis induced lymphopenia in 36 donors (47%) and monocytopenia in 23 donors (39%). Lymphopenia before first apheresis and prior history of PBSC harvest were independent factors of apheresis-induced lymphopenia while those factors had no influence on monocytopenia. A time-dependent decrease in lymphocyte counts was observed in donors undergoing repeated aphereses, resulting in persistent and prolonged lymphopenia in 50% of donors. No persistent monocytopenia over time and aphereses was observed. Kaplan-Meier estimate of the risk to develop persistent lymphopenia after multiple aphereses was 21% +/- 6% at 2 months, 38% +/- 8% at 4 months, and 64% +/- 10% at 12 months. After Cox regression analysis, previous PBSC harvest remained the unique factor associated with the risk for persistent lymphopenia. CONCLUSIONS: Monitoring the potential long-term effects of repeated aphereses in hematopoietic stem cell donors appears important. Selecting young bone marrow donors for subsequent DLI significantly reduces the risk for acute and prolonged lymphopenias. PMID- 15539081 TI - Estimating human hematopoietic stem cell kinetics using granulocyte telomere lengths. AB - OBJECTIVE: To study in vivo behavior of hematopoietic stem cells (HSC). MATERIALS AND METHODS: Behavior of HSC is difficult to study because one cannot observe and track cells within the marrow microenvironment. Therefore, information must be obtained from indirect means, such as competitive repopulation assays or surrogate studies, such as observations of telomere shortening in granulocytes. In this article, we use granulocyte telomere length data and a novel approach, stochastic simulation, to derive replication rates of HSC. The approach is first applied to cats and then to humans. RESULTS: Human HSC replicate infrequently, on average once per 45 weeks (range: once per 23 to once per 67 weeks). CONCLUSIONS: This rate is substantially slower than the average replication rates estimated for murine (once per 2.5 weeks) and feline (once per 8.3-10 weeks) HSC in vivo. PMID- 15539082 TI - Phosphatidylinositol-3-kinase activation mediates proline-rich tyrosine kinase 2 phosphorylation and recruitment to beta1-integrins in human CD34+ cells. AB - OBJECTIVE: beta1-integrins mediate hematopoietic stem and progenitor cell homing and retention in the bone marrow (BM) and inhibit hematopoietic proliferation and differentiation. Having no intrinsic kinase activity, integrins recruit intracellular kinases, such as the focal adhesion kinase (FAK) or the related proline-rich tyrosine kinase 2 (PYK2), to initiate signal transduction. Phosphatidylinositol-3-kinase (PI3K), which is involved in beta1-integrin signaling in many cell types, is physically and functionally associated with FAK in anchorage-dependent cells. Because PYK2 is the principal focal adhesion kinase expressed in primary human CD34+ cells, we assessed its functional relationship with PI3K in CD34+ cells in response to integrin engagement. METHODS: beta1 integrins on primary mobilized peripheral blood CD34+ cells and CD34+ KG1A cells were engaged by adhesion to fibronectin (FN) or by cross-linking with an anti beta1 integrin antibody, respectively. PI3K activity and PYK2 phosphorylation were then assessed in the presence or absence of the PI3K inhibitor, wortmannin. Association between PI3K, PYK2, and the beta1-integrin subunit were also evaluated in co-immunoprecipitation experiments. RESULTS: beta1-integrin engagement induced PI3K activation, which was required for, and temporally preceded, PYK2 phosphorylation, indicating that PI3K lies upstream of PYK2 in CD34+ cells. Furthermore, although PYK2 and PI3K were constitutively associated, interaction of the PYK2/PI3K complex with beta1-integrins required prior integrin engagement and PI3K activation. CONCLUSION: Activation of PI3K following beta1 integrin engagement on human CD34+ cells results in subsequent phosphorylation of PYK2, and is required for the recruitment of the PI3K/PYK2 complex to beta1 integrins at the cell surface. PMID- 15539083 TI - A novel mechanism for imatinib mesylate (STI571) resistance in CML cell line KT 1: role of TC-PTP in modulating signals downstream from the BCR-ABL fusion protein. AB - OBJECTIVE: Acquired resistance to imatinib mesylate (STI571) in chronic myelogenous leukemia (CML) patients has become a serious clinical problem. We previously established STI571-resistant sublines (designated KTR cells) from the CML cell line KT-1. T cell protein tyrosine phosphatase (TC-PTP) was markedly downregulated in all KTR cells compared to parental KT-1 cells. Therefore, we examined whether the suppression of TC-PTP expression might contribute to imatinib mesylate-resistance in KTR cells. MATERIALS AND METHODS: We transduced the nuclear isoform of TC-PTP (TC45) and catalytically inactive TC45-D182A cDNAs into KTR cells by retroviral gene transfer. Subsequently, we analyzed the sensitivity to imatinib mesylate and the status of signaling pathways in the transduced cells. RESULTS: The overall levels of STAT5 phosphorylation were significantly higher in KTR cells as compared to KT-1 cells, but reconstitution of TC-PTP in KTR cells resulted in a dramatic decrease of STAT5 phosphorylation. Furthermore, STAT5 phosphorylation was ablated by imatinib mesylate in KT-1 cells but remained elevated in KTR cells. In contrast, we observed no difference in BCR ABL or JAK2 phosphorylation and no difference in activation of other signaling pathways. Importantly, reconstitution of TC-PTP in KTR cells to levels found in parental KT-1 cells restored their sensitivity to imatinib mesylate as monitored by reduced proliferation and increased apoptosis. CONCLUSIONS: We have demonstrated that forced expression of TC-PTP in imatinib mesylate-resistant KTR cells can restore sensitivity to imatinib mesylate. Our studies indicate that loss of TC-PTP may represent a novel mechanism by which CML cells can acquire imatinib mesylate-resistance. PMID- 15539084 TI - Visualization of 5T33 myeloma cells in the C57BL/KaLwRij mouse: establishment of a new syngeneic murine model of multiple myeloma. AB - OBJECTIVE: Lack of good models for in vivo detection of multiple myeloma (MM) cells hampers our understanding of the disease. Our objective was to establish a murine model for MM, allowing sensitive and labor-free tracing and quantification of MM cells in an immunocompetent host. METHODS: 5T33MM cells were retrovirally transduced, expressing enhanced green fluorescent protein (eGFP) and/or herpes simplex virus thymidine kinase (HSV-tk) as a control. Flow cytometric eGFP detection accuracy and sensitivity were assessed. Functional characteristics of transduced cells, including growth rate and production of IgG2b paraprotein and interleukin-6, were compared to those of nontransduced cells in vitro. For induction of MM, C57BL/KaLwRij mice were injected intravenously with transduced and nontransduced cells. Survival kinetics and distribution of eGFP cells in tissues were evaluated. RESULTS: Flow cytometric eGFP detection was accurate at 1:1000 transduced/nontransduced cell ratio. Transduced and nontransduced 5T33MM cells exhibited similar growth rates, producing comparable IgG2b and interleukin 6 levels. Intravenous injection of both nontransduced and eGFP-transduced MM cells to C57BL/KaLwRij mice resulted in paraplegia. At the time of paraplegia, eGFP-transduced MM cells were detected substantially in the bone marrow, spleen, and liver, less in lymph nodes, but not in the thymus. The bone marrow of paraplegic mice contained higher eGFP-transduced MM cells compared to that of nonparaplegic animals. CONCLUSIONS: In the established eGFP-5T33 MM model, MM cells are easily traced in an immunocompetent host. This model simplifies the analysis of homing pattern studies, the evaluation of therapeutic effects of various treatment approaches and contributes towards better understanding of MM. PMID- 15539085 TI - Expression of TNF-related apoptosis-inducing ligand (TRAIL) in megakaryocytes and platelets. AB - OBJECTIVE: Platelets are known to play an important role in hemostasis, thrombosis, wound healing, and inflammation. Platelet-induced modulation of inflammation and adaptive immune responses are mediated in part through tumor necrosis factor (TNF) family member ligands, including CD154, Fas ligand, and TNFalpha, that are expressed upon platelet activation. The present study investigated whether platelets and megakaryocytes also express TNF-related apoptosis-inducing ligand (TRAIL), another pro-apoptotic member of the TNF superfamily. MATERIALS AND METHODS: Immunoprecipitation, enzyme-linked immunosorbent assay, and flow cytometry were used to assess TRAIL protein expression on isolated platelets, in vitro-derived megakaryocytes and premegakaryocyte cell lines. Reverse-transcription polymerase chain reaction and transient transfection of TRAIL promoter/reporter constructs were used to elucidate mechanisms of TRAIL regulation during megakaryocyte differentiation. TRAIL-dependent cytotoxicity assays were performed to determine if platelet derived TRAIL induces apoptosis of TRAIL sensitive target cells. RESULTS: Activated platelets expressed both membrane-bound and soluble TRAIL. TRAIL was also expressed by megakaryocytes, and in vitro studies showed that TRAIL expression was induced upon megakaryocyte differentiation. TRAIL expression was mediated by increased transcriptional activity of the TRAIL promoter, suggesting lineage-specific regulation of TRAIL during megakaryocyte differentiation. Abundant detergent-extractable, full-length TRAIL protein was observed in the lysates of platelets and megakaryocytes, but only low concentrations of TRAIL were released by nondetergent extraction methods. CONCLUSION: The data reported herein show that platelets express TRAIL that is synthesized by megakaryocytes and was expressed by activated platelets. While these data expand the spectrum of TNF family proteins expressed in platelets, the function of platelet-derived TRAIL is not known. PMID- 15539086 TI - Parvovirus B19 capsid protein VP2 inhibits hematopoiesis in vitro and in vivo: implications for therapeutic use. AB - OBJECTIVE: To evaluate the capacity of parvovirus B19 capsid protein VP2 to inhibit hematopoiesis in vitro and in vivo. If effective, a VP2-derived construct may have therapeutic and prophylactic utility in diseases associated to overproduction of hematopoietic cells. METHODS: The effect on hematopoiesis in vitro of recombinant VP2, intact and enzymatically fragmented, was evaluated in a colony formation assay, using cells from fetal liver and macaque bone marrow. VP2 was also administered intravenously in macaques and hematological parameters as well as the ex vivo colony formation were assayed during a follow-up period of 33 days. RESULTS: VP2 inhibited BFU-E colony formation by about 55%. CFU-GM and CFU GEMM colony formation was also affected. Fragmented VP2 retained the inhibitory effect. The ex vivo colony-forming capacity of macaque bone marrow cells was lower in animals that received VP2 injections, and a drop in hematocrit values was noted in one animal. CONCLUSION: VP2 has an inhibitory effect on hematopoiesis in vitro and in vivo. An active region within VP2 is implied, which would be a strong candidate for use as a medicament in diseases such as polycytemia vera. PMID- 15539087 TI - Low-dose radiation (LDR) induces hematopoietic hormesis: LDR-induced mobilization of hematopoietic progenitor cells into peripheral blood circulation. AB - OBJECTIVE: The aim of this study was to investigate the stimulating effect of low dose radiation (LDR) on bone marrow hematopoietic progenitor cell (HPC) proliferation and peripheral blood mobilization. METHODS: Mice were exposed to 25 to 100-mGy x-rays. Bone marrow and peripheral blood HPCs (BFU-E, CFU-GM, and c kit+ cells) were measured, and GM-CSF, G-CSF, and IL-3 protein and mRNA expression were detected using ELISA, slot blot hybridization, and Northern blot methods. To functionally evaluate LDR-stimulated and -mobilized HPCs, repopulation of peripheral blood cells in lethally irradiated recipients after transplantation of LDR-treated donor HPCs was examined by WBC counts, animal survival, and colony-forming units in the recipient spleens (CFUs-S). RESULTS: 75 mGy x-rays induced a maximal stimulation for bone marrow HPC proliferation (CFU GM and BFU-E formation) 48 hours postirradiation, along with a significant increase in HPC mobilization into peripheral blood 48 to 72 hours postradiation, as shown by increases in CFU-GM formation and proportion of c-kit+ cells in the peripheral mononuclear cells. 75-mGy x-rays also maximally induced increases in G CSF and GM-CSF mRNA expression in splenocytes and levels of serum GM-CSF. To define the critical role of these hematopoietic-stimulating factors in HPC peripheral mobilization, direct administration of G-CSF at a dose of 300 microg/kg/day or 150 microg/kg/day was applied and found to significantly stimulate GM-CFU formation and increase c-kit+ cells in the peripheral mononuclear cells. More importantly, 75-mGy x-rays plus 150 microg/kg/day G-CSF (LDR/150-G-CSF) produced a similar effect to that of 300 microg/kg/day G-CSF alone. Furthermore, the capability of LDR-mobilized donor HPCs to repopulate blood cells was confirmed in lethally irradiated recipient mice by counting peripheral WBC and CFUs-S. CONCLUSION: These results suggest that LDR induces hematopoietic hormesis, as demonstrated by HPC proliferation and peripheral mobilization, providing a potential approach to clinical application for HPC peripheral mobilization. PMID- 15539088 TI - CD8+ T cell dose affects development of acute graft-vs-host disease following reduced-intensity conditioning allogeneic peripheral blood stem cell transplantation. AB - OBJECTIVE: Acute graft-vs-host disease (aGVHD) remains an important cause of morbidity after reduced-intensity conditioning (RIC) allogeneic transplantation (allo-SCT). It has been shown that antithymocyte globulin (ATG) dose infused during RIC is a major determinant for the likelihood of developing aGVHD. The ATG modulation on aGVHD is likely related to in vivo T-cell depletion. PATIENTS AND METHODS: We therefore investigated the relationship between the cellular composition of the allograft and clinical outcome in 57 patients who received allogeneic peripheral blood stem cells from HLA-identical siblings following an ATG-based RIC. RESULTS: In a multivariate analysis, the CD8+ T cell dose infused was the only parameter associated with the risk of aGVHD (p=0.031; RR=1.96; 95% CI, 1.1-3.6). When looking at the extremes, patients experiencing grade III-IV aGVHD received a median of 143 x 10(6)/kg CD8+ T cells, while patients without aGVHD received a median of 96 x 10(6)/kg CD8+ T cells (p=0.021). None of the different cell subtypes contained in the allograft was associated with a significant probability of developing chronic GVHD. Patients with grade II aGVHD who received an intermediate dose of CD8+ T cells (median, 111 x 10(6)/kg) had a significantly better overall survival in comparison to patients with grade 0-I or grade III-IV aGVHD (p=0.009). CONCLUSION: In comparison to myeloablative allo SCT, these results demonstrate that a cautious monitoring of the number of cells infused, at least in the context of ATG-based RIC, may represent an important predictive indicator of early transplant-related events and outcome after RIC allo-SCT. PMID- 15539089 TI - rhG-CSF effect on mixed lymphocyte cultures and circulating soluble HLA antigen levels in volunteer stem cell donors. AB - OBJECTIVE: Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is a cytokine widely used in the procurement of peripheral blood stem cells (PBSC) from donors for allogeneic hematopoietic cell transplantation. Therefore, we were interested in its immediate and long-term effects on cellular and soluble factors known to be involved in the immune response. METHODS: We studied 35 PBSC donors by mixed lymphocyte culture (MLC) and lymphocyte transformation test (LTT), and 41 for soluble plasma factors (soluble human leukocyte antigen [sHLA]-G, -class I, -DR, and interleukin [IL]-10) pre and 5 days post initial rhG-CSF administration, respectively. In addition, 10 donors were reexamined at an average of 2 months (3-16 weeks) post-rhG-CSF. RESULTS: At 5 days post-rhG-CSF the donors presented a significant (p < 0.05) decrease of MLC, LTT mitogen, and recall antigen reactions. Plasma levels of sHLA-G, -class I, -DR, and IL-10 (p < 0.005 each) were significantly increased. The changes in IL-10 but not in sHLA were significantly (p < 0.05) correlated with LTT responses. In the 2-month follow-up there was no significant difference in alloreactivity and LTT reactions as compared to the pre-rhG-CSF results. The results generated after 3 to 16 weeks did not depend on the time point of investigation. Consistently, soluble factors decreased to pre-rhG-CSF levels. CONCLUSIONS: rhG-CSF administration suppresses cellular immune functions within 5 days and increases sHLA and IL-10 plasma levels. These immunomodulatory effects appear to be short-term only and vanished at an average of 2 months after rhG-CSF application. PMID- 15539090 TI - Depletion of donor-reactive cells as a new concept for improvement of mismatched bone marrow engraftment using reduced-intensity conditioning. AB - OBJECTIVE: New nonmyeloablative strategies to improve acceptance of mismatched bone marrow (BM) may compensate for the inadequate supply of compatible grafts. Recently we proposed to facilitate engraftment of mismatched BM by selective depletion of activated donor-reactive host cells with cyclophosphamide (CY). Here we have compared engraftment of allogeneic BM after depletion of antigen activated host lymphocytes by CY, with BM engraftment following general immunosuppression by the same CY dose. MATERIALS AND METHODS: Naive or mildly irradiated BALB/c mice were primed with C57BL/6 BM cells (day 0), treated with CY in order to deplete activated T cells (day 1), and transplanted with a second C57BL/6 BM inoculum (day 2) in order to achieve BM engraftment. Alternatively, mice received an equal dose of donor BM cells in a single injection one day after the same CY dose. Treated animals were repeatedly tested for persistence of donor cells in the blood. RESULTS: Depletion of alloantigen-primed lymphocytes by 200 mg/kg CY provided stable GVHD-free engraftment of allogeneic BM in nonirradiated mice, while immunosuppressive treatment with the same CY dose alone resulted in BM rejection. Low-dose irradiation before priming with donor BM allowed the tolerance-inducing CY dose to be reduced to 100 mg/kg. Alloantigen-primed lymphocyte depletion (APLD) by a reduced CY dose resulted in engraftment of donor BM after a significantly lower irradiation dose than treatment with irradiation and CY alone. CONCLUSION: Our results demonstrate that conditioning that focuses on APLD has a definite advantage over general immunosuppression with CY and radiation therapy. PMID- 15539091 TI - Enhanced engraftment of human cells in RAG2/gammac double-knockout mice after treatment with CL2MDP liposomes. AB - OBJECTIVE: The ability of human cells to repopulate the bone marrow of nonobese diabetic immunodeficient mice (NOD/SCID) is commonly used as a standard assay to quantify the primitive human hematopoietic stem cell population. We studied the applicability of the immunodeficient RAG2(-/-)gammac(-/-) double-knockout mouse for this purpose. METHODS: RAG2(-/-)gammac(-/-) mice and NOD/SCID mice were injected intravenously (i.v.) with umbilical cord blood-derived CD34(+) cells and engraftment was quantified by determining the human CD45+ cell chimerism in bone marrow at several time points. RAG2(-/-)gammac(-/-) were pretreated with total body irradiation and depleted of macrophages in liver, spleen, and bone marrow by i.v. injection of clodronate diphosphonate containing liposomes. RESULTS: We demonstrated that the frequency of chimerism and the level of engraftment in macrophage-depleted RAG2(-/-)gammac(-/-) largely resemble that in NOD/SCID mice. Also similar is the multilineage differentiation pattern in the two mouse strains at 7 weeks after transplantation, with a prominent outgrowth in RAG2(-/-)gammac( /-) of CD19+ cells (88% +/- 10%). Cells of other lineages were clearly less frequent: 9% +/- 2% myeloid cells and 0.1% +/- 0.1% erythroid cells. As for immature progenitors, 6% +/- 1% of the human cells express the CD34 antigen and 0.4% +/- 0.1% have the CD34+,CD33,38,71(-) phenotype. The presence of human committed progenitors (i.e., CFU-GM/BFU-E) was evident. The persistence of human cells at 4 months after transplantation shows that the RAG2(-/-)gammac(-/-) support long-term maintenance of human hematopoiesis. CONCLUSION: Our findings indicate that macrophage-depleted RAG2(-/-)gammac(-/-) are a suitable model for studying human hematopoiesis including multipotential stem cells, and long-term repopulation. PMID- 15539092 TI - Imaging of the elbow following trauma. PMID- 15539093 TI - Rehabilitation of elbow trauma. AB - The rehabilitation of elbow trauma presents numerous challenges. Involvement of the osseous structures, compromise of the ligamentous stability, and loss of the soft tissue excursion necessary for elbow motion and function require due consideration during the treatment of elbow joint injuries. Stiffness of the elbow joint following trauma is common. This stiffness is caused by extrinsic and intrinsic factors. Contractures of the elbow joint develop as a result of contractures of the joint capsule, ligamentous structures, musculotendinous structures, intra-articular adhesions,and ectopic ossification. Early mobilization and splinting of the elbow following injury, within a safe arc of elbow motion, makes the elbow joint more compliant to the rehabilitative techniques outlined. Understanding the details of elbow anatomy, biomechanics, trauma, and surgical procedures for repairing the osseous and the ligamentous structures thus are the key factors in rehabilitating the elbow joint successfully. PMID- 15539094 TI - Surgical approaches to the elbow. AB - Success in elbow surgery depends on a thorough understanding of its anatomy and the access to its various compartments and components.This article has reviewed the current applied surgical anatomy of the elbow and the related surgical approaches. For acute elbow injuries the Global approach is preferred, because this incorporates a posterior skin incision to expose the medial and lateral sides of the joint. The "Z"arthrotomy algorithm avoids PLRI when exposing the lateral joint capsule. The acute distal biceps tendon avulsion can be repaired with a minimally invasive anterior approach. PMID- 15539095 TI - Simple dislocations of the elbow: evaluation and treatment. AB - Simple dislocations of the elbow or dislocations occurring without an associated fracture are common injuries. Evaluation of these injuries must include an assessment of the entire involved upper extremity and a complete neurovascular examination. Principles of management include a prompt, controlled reduction, a determination of postreduction stability, and an immediate rehabilitation protocol that considers the stability of the joint following reduction. For those joints that are stable throughout the arc of motion, an unrestricted range of motion protocol can be started. When instability is present after reduction, the degree of instability determines the need for dynamic bracing, ligament repair or reconstruction, or the need for a hinged external fixator. The long-term results of these injuries are generally good, with nonoperative treatment producing equivalent or better results than operative treatment. Residual flexion contracture is the most common complication and can be diminished with the use of early range of motion. Other common complications include residual pain and heterotopic ossification, whereas recurrent instability occurs infrequently. Chronic dislocations of the elbow occur uncommonly in North America and Western Europe, but when they occur, reason-able results can be achieved with open reduction of the joint and the use of a hinged external fixator within I year following the dislocation. PMID- 15539096 TI - Fracture-dislocation of the elbow: diagnosis, treatment, and prognosis. AB - There are two main goals for the surgeon treating injuries around the elbow joint: the maintenance of a stable, concentric reduction of the joint and the initiation of early motion to help prevent stiffness and maximize functional out come. The presence of a fracture decreases joint stability and increases the risk for early subluxation or dislocation with motion. Operative repair thus is indicated for most of these injuries to restore sufficient osseoligamentous support to allow safe, early motion and provide a stable functional elbow in the long term. PMID- 15539097 TI - Open reduction and internal fixation of fractures of the radial head. AB - Early reports of open reduction and internal fixation of fractures of the radial head were positive, perhaps because of the prevalence of isolated partial head fractures for which good results would be expected. Subsequent reports have found that complex fractures of the radial head are prone to early failure, nonunion, and poor forearm rotation after operative fixation. Combined with increased availability and use of metal radial head prosthesis for complex fractures of the radial head, the role of open reduction and inter-nal fixation is being redefined. PMID- 15539098 TI - Management of comminuted radial head fractures with replacement arthroplasty. AB - Radial head arthroplasty is indicated for displaced comminuted radial head fractures that cannot be managed reliably with open reduction and internal fixation and that have an associated elbow dislocation. Replacement also is indicated in patients with comminuted radial head fractures that have or are likely to have a disruption of the medial col-lateral, lateral collateral, or interosseous ligaments. Biomechanical studies have demonstrated that metallic implants restore elbow stability similar to the native radial head. The early and midterm clinical experience with metallic radial head arthroplasty has been encouraging relative to earlier reports with silicone devices. Newer modular designs incorporate improved sizing to better reproduce the anatomy of the proximal radius and are easier to insert intraoperatively. PMID- 15539099 TI - Fractures of the coronoid process. AB - Proper treatment of coronoid fractures requires an understanding of the bony and soft tissue anatomy of the elbow and the various injury mechanisms that occur. Newer fracture classifications help direct treatment based on the size and location of the fracture, injury mechanism, and associated stability of the elbow. PMID- 15539100 TI - Coronal shear fractures of the distal humerus: the capitellum and trochlea. AB - Fractures of the capitellum and trochlea are uncommon and multiple options have been advocated for the treatment of this injury. A single management technique has not emerged as the superior technique, and a complement of interventions is necessary to manage the continuum of injuries that can be observed. In general, open reduction and internal fixation is advocated for healthy and active patients with satisfactory bone quality to allow for the insertion of stable fixation. In the geriatric population, total elbow arthroplasty may emerge as the treatment of choice particularly for the more comminuted fracture patterns. Postoperative rehabilitation is important and is guided by fracture stability, ligament integrity, and the ability of the patient to cooperate with the treatment protocol. Gratifying results can be achieved in most patients with even the most complex injuries. PMID- 15539101 TI - Supracondylar fractures of the elbow: open reduction, internal fixation. AB - This principle-based approach for fixing distal humerus fractures has many advantages. Complex fractures are able to be fixed with sufficient stability to permit immediate intensive rehabilitation. Some fractures believed to be unfixable have been fixed satisfactorily by applying the principles outlined in this article. More straightforward fractures are fixed easily using the same techniques. In the author's experience, the stability achieved with this approach is so much greater than that with traditional methods of fixing distal humerus fractures that bone graft has been required only rarely, despite the severity of injuries so typical of the tertiary referral nature of the author's practice. The key points are that the plates should be placed in parallel configuration medially and laterally and that the screws passing through these plates in the distal fragments should interdigitate and lock together. PMID- 15539102 TI - Total elbow anthroplasty and distal humerus elbow fractures. AB - Palvanen et al in 2003 collected data on osteoporotic fractures of the distal humerus fractures from the Finnish National Hospital Discharge Register. They defined an osteoporotic fracture of the distal humerus as a fracture occurring in persons 60 years of age or older following moderate or minimal trauma. The annual rate of incidence of these types of fractures has increased significantly, with 42 fractures reported in 1970 and 208 fractures in 2000(395% increase). The age specific incidence rates showed a ninefold increase for women 80 years of age or older (8 in 1970 versus 75 in 2000). In the 60-69-year and 70-79-year age groups, the age adjusted increases were twofold. This presents great challenges for future orthopedists, reinforcing the ongoing need to critically analyze results of treatment for this difficult problem. Many patients over the age of 60 years can do well with traditional open reduction and internal fixation of distal humerus fractures. When patients have poor bone quality, however, preventing stable internal fixation or significant medical comorbidities, TEA should be considered. Encouraging results have been reported using TEA as a primary mode of treatment for carefully selected comminuted distal humerus fractures in elderly patients, particularly if the patient also has significant rheumatoid arthritic changes. A triceps-sparing approach is recommended so that patients may be mobilized early following surgery. PMID- 15539103 TI - Arthroscopic management of elbow trauma. AB - The arthroscope can assist in effectively treating traumatic injuries about the elbow, including some intra-articular fractures. Proper selection of patients is important to ensure effective treatment, but when used judiciously, arthroscopic assisted debridement and fracture fixation is effective. The arthroscope is a valuable tool in the diagnosis and management of acute intra-articular fractures and in dealing with the sequelae that sometimes cause problems after healing has occurred. It allows for effective fracture treatment without the extensive soft tissue disruption that would be otherwise necessary for exposure. The authors certainly cannot advocate arthroscopic treatment of all intra-articular elbow fractures, but in specific circumstances, the arthroscope cannot only assist but also improve treatment of these sometimes difficult and often challenging injuries. The arthroscope is also useful in the identification of varus, valgus, and posterolateral rotatory instability, particularly when these instabilities are less obvious on clinical examination. PMID- 15539105 TI - Should albuminuria be a therapeutic target in patients with hypertension and diabetes? AB - Conventional risk factors associated with cardiovascular and renal complications of diabetes include hypertension, dyslipidemia, hyperglycemia, and smoking. Recently, albuminuria has also emerged as an important risk factor, as it is independently associated with increased cardiovascular and renal risk. Inhibition of the renin-angiotensin-aldosterone system (RAAS) reduces both blood pressure (BP) and albuminuria, and induced cardiovascular and renal protection is associated with this albuminuria reduction, independent of BP reduction. Based on these results, optimal therapy for patients at risk for type 2 diabetic nephropathy should include BP reduction to <130/80 mm Hg, with the inclusion of a RAAS blocking agent (ie, an angiotensin II receptor blocker) to provide BP control and control of urinary albumin, which should be reduced to <500 mg/d. PMID- 15539106 TI - The role of the renin-angiotensin-aldosterone system in diabetes and its vascular complications. AB - Clinical trials have demonstrated the benefit of blood pressure (BP) reduction in reducing the risk of cardiovascular and renal complications in patients with diabetes mellitus. Incorporation of agents that inhibit the renin-angiotensin aldosterone system (RAAS) into antihypertensive regimens has been shown to provide reductions in renal and cardiovascular events that are mediated by both BP-dependent and -independent mechanisms. Recent studies exploring these potential mechanisms have demonstrated a direct role of angiotensin II (ATII) in the pathology of the vasculature and other sites of end-organ injury. In animal models of diabetes, inhibition of the RAAS with angiotensin-converting enzyme (ACE) inhibition or angiotensin type 1 (AT(1)) receptor blockade has been shown to prevent atherosclerosis, an effect that was independent of BP reduction. In addition to its direct effects on the vasculature, ATII also has direct detrimental effects on end organs, including the kidney and the heart, which lead to the development of proteinuria and left ventricular hypertrophy (LVH), respectively. Left ventricular hypertrophy has been shown to be predictive of cardiovascular and renal events, and the benefits of RAAS inhibition with angiotensin receptor blocker therapy are accompanied by a reduction in LVH. In addition to preventing the cardiovascular and renal complications of diabetes, the RAAS blockade has also been shown, in several large randomized clinical trials, to inhibit new onset of diabetes. Recent studies have revealed that many tissues, including pancreatic islets and adipose tissue, have a local RAAS. In the diabetic rat model (Zucker diabetic fatty rats), pancreatic islets exhibit an increased intraislet expression of ACE and AT(1) as well as increased intraislet fibrosis, apoptosis, and oxidative stress. The local RAAS also appears to play a role in the function of the adipocyte. Angiotensin II inhibits adipocyte differentiation, potentially decreasing the storage capacity of adipose tissue. The reduced capacity of adipose tissue to store fatty acids may cause their accumulation in other tissues, leading to insulin resistance and development of diabetes. Collectively, these studies demonstrate that ATII has direct effects on multiple tissues, and inhibition of ATII action in these tissues may be responsible for many of the clinical benefits observed with RAAS inhibition. PMID- 15539107 TI - Insights into the biology of diabetic vascular disease: what's new? AB - The major cause of morbidity and mortality in persons with diabetes is cardiovascular disease (CVD), the risk of which is increased three- to four-fold versus persons without diabetes. The biology of diabetes is characterized not only by hyperglycemia but also by hypertension, dyslipidemia, microalbuminuria, inflammation, and abnormal thrombolysis. Hypertension is a common feature of diabetes and is the primary contributor to CVD. Recent investigations have revealed a relationship between vascular derangements, insulin resistance, and visceral obesity and have implicated the renin-angiotensin-aldosterone system (RAAS) as a key mediator of cardiovascular dysfunction in diabetes. Angiotensin II has been shown to have direct effects on endothelial dysfunction, oxidative stress, inflammation, skeletal muscle, and adipocyte function. These pathophysiologic considerations have formed the basis for CVD prevention strategies in diabetes. Clinical trials have demonstrated a reduction in cardiovascular events with aspirin, lipid-lowering agents, and antihypertensive agents. Blood pressure (BP) control (<130/80 mm Hg) is a crucial component of risk reduction, and several studies have demonstrated the need for multiple agents to reach therapeutic goals. Clinical trials also demonstrated the benefit of RAAS blocking agents in reducing BP and cardiovascular and renal risk, and suggest clinical benefits beyond BP reduction. PMID- 15539108 TI - Lessons learned from recent clinical trials in hypertensive diabetics: what's good for the kidney is good for the heart and brain. AB - Hypertension and interactions of the activated renin-angiotensin-aldosterone system (RAAS) are major risk factors for both cardiovascular disease and renal disease in patients with diabetes. Recent clinical trials show that specific inhibition of the RAAS may significantly decrease morbidity and mortality from cardiovascular and renal causes in this high-risk population and reduce risk for new-onset diabetes in patients with hypertension. In addition, these trials demonstrated that benefits of RAAS blockade are mediated by both BP-dependent and -independent effects, and they suggest that the cardiovascular benefits of RAAS blockade may result from its favorable effects on both cardiac and renal function. PMID- 15539109 TI - Brd4: tethering, segregation and beyond. AB - Papillomaviruses segregate their genomes in dividing cells by tethering them to mitotic chromosomes via the viral E2 protein. A recent report has shown that this interaction is mediated by the cellular bromodomain protein Brd4. This discovery provides new insight into the mechanism of viral genome segregation and raises many exciting questions about the regulation and nature of the interaction of this complex with mitotic chromosomes. PMID- 15539110 TI - Battling enteroinvasive bacteria: Nod1 comes to the rescue. AB - Recognition of pathogenic bacteria by mammalian hosts is largely mediated by membrane-bound Toll-like receptors (TLRs). Recently, a family of cytosolic proteins, termed NODs, with homology to plant disease-resistance gene products has been implicated in sensing microbes within the cytosol. The role of NOD family members in host defense is largely unknown. However, a recent report revealed that Nod1 is a crucial sensor for certain enteroinvasive bacteria that avoid TLR signaling. This finding suggests that Nod1 plays an important role in the initial recognition of pathogenic bacteria at epithelial surfaces, such as the gut, where innate immune responses to commensal bacteria must be avoided. PMID- 15539111 TI - It's a cold world out there (but the prospects are hot). PMID- 15539112 TI - Rocking the curve. PMID- 15539113 TI - What do microarrays really tell us about M. tuberculosis? AB - Bacterial pathogens adapt to their host environments to a large extent through switching on complex transcriptional programmes, and whole-genome microarray experiments promise to reveal this complexity. There has been a recent burst of articles reporting transcriptome analyses of Mycobacterium tuberculosis, including for the first time studies in macrophages and mice. We review gene expression reports, and compare them with each other and with microarray-based gene essentiality studies, revealing at times a startling lack of correlation. Additionally, we suggest a standardization format for the submission of processed data for publication, to facilitate cross-experiment analyses. PMID- 15539114 TI - Evolution of polydnaviruses as insect immune suppressors. AB - Polydnaviruses (PDVs) are endogenous particles that are used by some endoparasitic hymenoptera to disrupt host immunity and development. Recent analyses of encapsidated PDV genes have increased the number of known PDV gene families, which are often closely related to insect genes. Several PDV proteins inactivate host haemocytes by damaging their actin cytoskeleton. These proteins share no significant sequence homology and occur in polyphyletic PDV genera, possibly indicating that convergent evolution has produced functionally similar immune-suppressive molecules causing a haemocyte phenotype characterised by damaged cytoskeleton and inactivation. These phenomena provide further insights into the immune-suppressive activity of PDVs and raise interesting questions about PDV evolution, a topic that has puzzled researchers ever since the discovery of PDVs. PMID- 15539115 TI - Surface polysaccharides enable bacteria to evade plant immunity. AB - Plants have an immune system to perceive pathogenic or potentially beneficial bacteria. Aspects of perception, signal transduction and the responses that the plant produces resemble features of innate immunity observed in animals. Plant reactions are various and include the production of antimicrobial compounds. Bacteria that are successful in establishing pathogenic or symbiotic interactions have developed multiple ways to protect themselves. We review the general importance of bacterial surface polysaccharides in the evasion of plant immune responses and elaborate on their role in protecting symbiotic bacteria against toxic reactive oxygen species during invasion of the host plant. PMID- 15539116 TI - Does the microbiota regulate immune responses outside the gut? AB - Perturbations in the gastrointestinal (GI) microbiota composition that occur as a result of antibiotics and diet in "westernized" countries are strongly associated with allergies and asthma ("hygiene hypothesis"). The microbiota ("microflora") plays a crucial role in the development of mucosal tolerance, including the airways. Significant attention has been focused on the role of the microbiota in GI development, immune adaptation and initiation of GI inflammatory diseases. This review covers the post-developmental functions that the microbiota plays in regulating immunological tolerance to allergen exposure outside the GI tract and proposes the question: is the microbiota a major regulator of the immune system? PMID- 15539117 TI - Receptor clustering and signal processing in E. coli chemotaxis. AB - Chemotaxis in Escherichia coli is one of the most thoroughly studied model systems for signal transduction. Receptor-kinase complexes, organized in clusters at the cell poles, sense chemoeffector stimuli and transmit signals to flagellar motors by phosphorylation of a diffusible response regulator protein. Despite the apparent simplicity of the signal transduction pathway, the high sensitivity, wide dynamic range and integration of multiple stimuli of this pathway remain unexplained. Recent advances in computer modeling and in quantitative experimental analysis suggest that cooperative protein interactions in receptor clusters play a crucial role in the signal processing during bacterial chemotaxis. PMID- 15539118 TI - Transplantation of skeletal myoblasts for cardiac repair. PMID- 15539119 TI - Previous thoracic surgery does not increase peri-operative mortality in pediatric heart-lung transplant recipients. AB - BACKGROUND: Heart-lung transplant (HLT) is indicated in select children with end stage cardiopulmonary disease. We sought to determine whether previous thoracic surgery increases peri-operative morbidity and mortality. METHODS: Retrospective data were analyzed using unpaired Student's t-test and Fisher's exact test. Results are reported as mean +/- SD. Peri-operative mortality was defined as death at 4 hours had the highest association with 30-day mortality, followed by CrCl 50 ml/min (19.7% vs 9.5%; p < 0.01). The change in CrCl between registration and transplantation was not related to mean CrCl or waiting time. In 67 of the patients with a CrCl >50 ml/min at registration, CrCl was reduced to 1 year after HTX) during a 5-year follow-up using the Medication Event Monitoring System (MEMS) for compliance assessment. METHODS: We categorized 101 patients (86% men; aged 55 +/- 10 years; time after HTX at inclusion, 1,253 +/- 534 days) as medication non-compliers (n = 17) or compliers (n = 84) based on the MEMS. Late acute rejections, transplant coronary artery disease (TxCAD), retransplantation, and death were registered during the 5-year follow-up. RESULTS: Non-compliers had significantly more TxCAD (p = 0.025). Non-compliers also had a greater rate of late acute rejection (11.8% vs 2.4%) and retransplantation (13.3% vs 2.5%)), although these differences were not statistically significant. Mortality rates were similar. Kaplan Meier analysis showed that non-compliers had a significantly shorter clinical-event free time compared with compliers (mean, 1,318 vs 1,612 days; p = 0.043). Cox regression analysis showed that the adjusted relative risk associated with non compliance was 2.03 (p = 0.0582), after controlling for other known transplant related risk factors for poor clinical outcome. CONCLUSION: The current study demonstrates that medication non-compliance in the late post-transplant period doubles the risk for an untoward clinical event. Non-compliance is a continuous risk factor after heart transplantation that should be targeted by compliance enhancing interventions. PMID- 15539123 TI - Donor-recipient gender mismatch in lung transplantation: impact on obliterative bronchiolitis and survival. AB - BACKGROUND: Because of the shortage of donor lungs, liberalization of donor selection criteria in terms of age, gas exchange, and smoking history has been proposed. METHODS: We evaluated a single-institution population of lung transplant recipients (n = 98) for donor-recipient gender matching. We measured overall survival, time to acute allograft rejection, and time to development of obliterative bronchiolitis (OB). RESULTS: We found significant improvement in overall survival for gender-mismatched donor and recipient pairs (p = 0.078) and a significantly shorter OB-free period for male donor and female recipient pairs (p = 0.017). CONCLUSION: These findings suggest that donor organ allocation based on gender may affect long-term survival and other outcomes after lung transplantation. PMID- 15539124 TI - Increased plasma homocysteine concentrations accelerate cardiac allograft vasculopathy. AB - BACKGROUND: A toxic and pro-oxidative effect of homocysteine on the coronary endothelium may accelerate cardiac allograft vascular disease (CAVD). In this study, we evaluated the influence of hyperhomocysteinemia on the course of CAVD. METHODS: We investigated plasma homocysteine (tHCY) concentrations in 183 consecutive heart transplant recipients (158 men and 25 women, mean aged 53.1 +/- 10.0 years, at 6.7 +/- 3.2 years after transplantation) to evaluate the course of CAVD. We used serial coronary angiography to assess coronary status and graded the severity of CAVD based on the extent of luminal obstruction in the main coronary arteries (graded as 1-4). We defined progression as increased focal stenosis of >/=30% or as detection of a new coronary lesion after a mean observation period of 2.8 +/- 1.0 years. A multivariate analysis (backward logistic regression) was performed that included potential risk factors for CAVD. We excluded patients undergoing dialysis. RESULTS: Initially, tHCY concentrations were increased in the entire cohort (mean, 18.6 +/- 7.6 mumol/liter) and ranged from 6.6 to 46.9 mumol/liter. A total of 105 patients (57.0%) had CAVD at first angiography, and progression was detected in 52 transplant recipients (28.0%). Patients with progressive CAVD had significantly greater tHCY concentrations (21.6 +/- 6.2 mumol/liter) at baseline investigation compared with patients who had stable courses (17.4 +/- 7.7 mumol/liter; p < 0.001). These results were independent of parameters such as sex, age, dyslipoproteinemia, cyclosporine blood concentrations, and indication for transplantation. CONCLUSIONS: Progression of CAVD is strongly associated with increased tHCY concentrations. The intervals between routine surveillance angiography should be shortened in patients with hyperhomocysteinemia, and routine medical treatment to decrease homocysteine concentrations should be considered. PMID- 15539125 TI - alpha4beta1-integrin blockade and cyclosporine decreases the prevalence and severity of transplant vasculopathy in a rat transplant model. AB - BACKGROUND: Transplant vasculopathy leads to neointimal proliferation of allograft arteries, and alpha4beta1-integrin (very late antigen-4 [VLA-4]) seems to play an important role in the pathogenesis. This study evaluates the effect of a new, synthetic, VLA-4 blocker (S3429) on transplant vasculopathy in a rat cardiac transplant model. METHODS: After transplantation (Lewis to Fisher), rats were divided randomly into 6 therapy groups: Group 1, n = 14, saline solution (vehicle); Group 2, n = 14, 3 mg/kg/day cyclosporine; Group 3, n = 21, 10 mg/kg/day S3429 + 3 mg/kg/day cyclosporine; Group 4, n = 21, 5 mg/kg/day S3429 + 3 mg/kg/day cyclosporine; Group 5: n = 21, 10 mg/kg/day S3429; Group 6, n = 21, 5 mg/kg/day S3429. Cyclosporine was given continuously until rats were killed. S3429 was either given for the entire study time or was discontinued after 20 days and animals were killed at Day 80. Twenty-eighty days after grafting, we assessed vasculopathy prevalence and mean vessel occlusion in coronary arteries. RESULTS: Cyclosporine decreased the prevalence of vasculopathy and mean vessel occlusion compared with controls. We observed a further decrease in prevalence and mean vessel occlusion with 80 days of therapy with S3429 and cyclosporine. After discontinuing S3429 therapy at Day 20, prevalence and mean vessel occlusion increased to values seen in cyclosporine-treated animals at Day 80. S3429 alone decreased mean vessel occlusion only within the first 20 days compared with controls but had no effect on the prevalence of vasculopathy. CONCLUSION: Because of the further decrease with S3429 therapy and the dramatic increase after discontinuation of S3429 therapy, we conclude that blocking VLA-4 receptors may prevent the development of transplant vasculopathy. PMID- 15539126 TI - Pre-emptive treatment with oral valganciclovir in management of CMV infection after cardiac transplantation. AB - BACKGROUND: Cytomegalovirus (CMV) has long been recognized as the most common opportunistic pathogen in transplant recipients. The use of post-detection antiviral treatment of CMV as a strategy to prevent disease in cardiac recipients is becoming the standard policy. Valganciclovir is an oral pro-drug of ganciclovir, with a 10-fold greater bioavailability than oral gancyclovir. PATIENTS AND METHODS: We reported our first experience with 8 patients (3 female, 45.0 +/- 10.5 years old, non-CMV mismatched) who underwent cardiac transplantation and had positive results of CMV polymerase chain reaction (PCR) within first 6 weeks after transplantation without concomitant CMV disease. These patients received valganciclovir in dosage 450 to 900 mg daily depending on renal function for 3 weeks. Standard immunosuppressive regimen consisted of cyclosporin A, MMF and corticosteroids, and was not changed after detection of CMV infection. In one patient we used sirolimus with respectively reduced dosage of cyclosporin A. Weekly measurements of CMV-PCR were performed to observe results of therapy. RESULTS: After 1 week of valganciclovir therapy CMV-PCR plasma concentration in all patients decreased significantly (2,105 copies/ml vs 400 copies/ml; p < 0.0001). No relapse of CMV infection has been detected after completing of valganciclovir therapy with follow up duration of 9.0 +/- 0.92 months. The drug was generally well tolerated, and we did not observe any severe drug related adverse events. CONCLUSION: Oral valganciclovir as pre-emptive antiviral therapy administrated after detection of CMV infection seems to be an effective and safe treatment in cardiac transplant recipients. PMID- 15539127 TI - Effect of left ventricular assist device combination therapy on myocardial blood flow in patients with end-stage dilated cardiomyopathy. AB - BACKGROUND: Changes in myocardial blood flow (MBF) and coronary flow reserve (CFR) are independent prognostic risk factors in idiopathic dilated cardiomyopathy (DCM). The aim of this study was to assess the impact of left ventricular unloading using left ventricular assist device (LVAD) combination therapy on resting MBF and CFR in patients with end-stage heart disease. METHODS: We studied 11 patients with deteriorating end-stage DCM (New York Heart Association Class 4) treated with LVAD support combined with pharmacologic therapy in a recovery program. Absolute MBF was measured using oxygen-15-labeled water (H(2)(15)O) positron emission tomography (PET) at rest during LVAD support and 15 minutes after the LVAD was switched off. Data were corrected for rate pressure product (RPP) when appropriate. Hyperemic MBF (intravenous adenosine, 140 mug/kg . min) was also measured in 6 patients with the LVAD switched off. CFR was calculated as the ratio MBF adenosine/MBF LVAD off (corrected). Data are expressed as mean +/- SD. RESULTS: At 317 +/- 193 days after device implantation, resting MBF was 0.95 +/- 0.29 (LVAD on) and 1.46 +/- 0.62 (LVAD off, corrected) ml/min . g (p = 0.01). MBF (LVAD on) was comparable with that of 11 age- and gender-matched normal controls (1.09 +/- 0.22 ml/min . g). CFR in the LVAD group was 1.49 +/- 0.99 compared with 3.56 +/- 1.42 in normal controls (p < 0.01). CONCLUSIONS: During LVAD support, resting MBF (LVAD on) was comparable to MBF in normal controls and increased when the LVAD was switched off. However, CFR was significantly impaired, even though all patients studied showed varying degrees of myocardial recovery. The implications of these findings, particularly in the long term, require further study. PMID- 15539128 TI - Role of poly (ADP) ribose synthetase in lung ischemia-reperfusion injury. AB - BACKGROUND: The activation of poly (adenosine diphosphate) ribose synthetase (PARS) is known to be important in the cellular response to oxidative stress. Previous studies have reported that PARS inhibition confers protection in models of endotoxic shock and ischemia-reperfusion. The purpose of this study was to determine the role of PARS inhibition in lung ischemia-reperfusion injury (LIRI). METHODS: Left lungs of Long-Evans rats were rendered ischemic for 90 minutes and reperfused for up to 4 hours. Treated animals received 3 mg/kg of INO-1001 (a PARS inhibitor) intravenously 30 minutes before ischemia. Injury was quantitated in terms of tissue myeloperoxidase (MPO) content, vascular permeability ((125)I radiolabeled bovine serum albumin extravasation) and bronchoalveolar lavage (BAL) leukocyte content. BAL fluid was assessed for cytokine and chemokine content by enzyme-linked immunoassay. Further samples were processed for nuclear protein analysis by electromobility shift assay (EMSA) and cellular death by terminal deoxyribonucleotidyl transferase-mediated d-UTP biotin nick-end labeling (TUNEL) assay and caspase-3 staining. RESULTS: Lung vascular permeability was reduced in treated animals by 73% compared with positive controls (p < 0.009). The protective effects of PARS inhibition correlated with a 46% reduction in tissue MPO content (p < 0.008) and marked reductions in BAL leukocyte accumulation. This positively correlated with the diminished expression of pro-inflammatory mediators and nuclear transcription factors, as well as decreased levels of cellular death. CONCLUSIONS: The deleterious effects of LIRI are in part mediated by the formation of free radicals and superoxides, which lead to DNA single strand breaks. This leads to activation of PARS, which causes rapid cellular energy depletion and cell death. PARS inhibition is protective against this and represents a potentially useful therapeutic tool in the prevention of LIRI. PMID- 15539129 TI - Adrenomedullin messenger RNA expression is increased in myocardial tissue of patients with idiopathic dilated cardiomyopathy. AB - Increased plasma levels of adrenomedullin (ADM) have been reported in patients with congestive heart failure Immunohistochemical ADM has been identified in failing human ventricle, but the gene expression pattern of ADM messenger RNA (mRNA) in myocardial tissue of patients with heart failure has not been elucidated. In this study, gene expression of ADM mRNA (analyzed by northern blot) and tissue concentration of ADM (measured by radioimmunoassay) were assessed in the explanted hearts of 17 patients with idiopathic dilated cardiomyopathy (IDC) and in 7 organ donors with no cardiopathy (controls). Myocardial tissue samples of patients with IDC showed increased ADM mRNA gene expression (p < 0.05) and decreased immunoreactive ADM protein content (p < 0.02) compared with controls. PMID- 15539130 TI - Electrocardiographic changes simulating a myocardial infarction after cardiac transplantation. AB - A 39-year-old man with idiopathic dilated cardiomyopathy and New York Heart Association Class III heart failure symptoms underwent orthotopic cardiac transplantation. Post-operatively, he developed an anteroseptal infarct pattern on the electrocardiogram (ECG), without regional wall motion abnormalities according to echocardiography. This pseudo-infarct pattern on ECG resolved within 6 months without coronary intervention or sequelae. It is postulated that these ECG changes were caused by increased left ventricular wall thickness due myocardial wall edema and reperfusion injury after cardiac transplantation. PMID- 15539131 TI - Cardiac transplantation: a temporary solution for Friedreich's ataxia-induced dilated cardiomyopathy. AB - Friedreich's ataxia is an autosomal recessive neurodegenerative disease. We report the case of a 34-year-old man with Friedreich's ataxia and dilated cardiomyopathy who underwent successful cardiac transplantation. To our knowledge, this is the first reported case of a heart transplantation for Friedreich's ataxia dilated cardiomyopathy. PMID- 15539132 TI - James D. Hardy: a pioneer in surgery (1918 to 2003). PMID- 15539133 TI - Cash injection for thermostable vaccines. PMID- 15539134 TI - A 'bioreactor' for bladder cancer cells. PMID- 15539135 TI - The fight against cancer: the next round. PMID- 15539136 TI - Ontology-based knowledge management of troglitazone-induced hepatotoxicity. PMID- 15539137 TI - Cite and oversight. PMID- 15539138 TI - Maurice Wilkins: the third man of the double helix. PMID- 15539139 TI - Intrabodies: production and promise. AB - Antibodies are among the most powerful tools in biological research and are presently the fastest growing category of new drug entities. It has long been a dream to harness their power to probe and modulate activities inside living cells. The binding of an antibody to an intracellular molecule has the potential to block, suppress, alter or even enhance the process mediated by that molecule. In particular, intracellular use of antibody fragments can offer an effective alternative to gene-based knockout technologies, potentially with more control and subtlety of outcome. This article outlines progress in the development of intracellular antibodies or intrabodies and highlights their potential, both as drug-discovery tools and as drug entities in their own right. PMID- 15539140 TI - Strategies for drug discovery by targeting sulfation pathways. AB - Posttranslational modifications of proteins such as phosphorylation have been recognized as pivotal modulators of biological activity in healthy and diseased tissues. Sulfation is a key posttranslational modification the role of which in physiology and pathology is only now becoming appreciated. Whereas phosphorylation is central to intracellular signal transduction, sulfation modulates cell-cell and cell-matrix communication. Sulfation involves a class of enzymes known as sulfotransferases, which transfer sulfate from the ATP-like sulfate donor 3'phosphoadenosine-5'phosphosulate to glycoproteins, glycolipids or metabolites. This review focuses on Golgi-localized sulfotransferases, their molecular biology and biochemistry, and strategies towards discovery of sulfotransferase inhibitors that could have potential as therapeutics in inflammation, cancer and infectious diseases. PMID- 15539141 TI - Molecular biomarkers in drug development. AB - Arguably, the most immediately promising reverberation of the genomics era has been the application of biomarkers to drug development. The promise of applying biomarkers to early drug development is that they might aid in preclinical and early clinical decisions such as dose ranging, definition of treatment regimen, or even a preview of efficacy. Later in the clinic, biomarkers could be used to facilitate patient stratification, selection and the description of surrogate endpoints. Information derived from biomarkers should result in a better understanding of preclinical and clinical data, which ultimately benefits patients and drug developers. If the promise of biomarkers is realized, they will become a routine component of drug development and companions to newly discovered therapies. PMID- 15539146 TI - Antigen-driven T cell expansion: affinity rules. AB - TCR affinity and ligand off-rate have both been found to influence the degree of T cell activation by peptide-MHC. A report in this issue of Immunity finds that ligand off-rate does not correlate with antigen-specific peripheral T cell expansion. Moreover, the data point to the surprising notion that there exist TCR affinity thresholds and, once attained, T cells with higher affinity receptors have no competitive advantage. PMID- 15539147 TI - Fingering IL-12 with apoptotic cells. AB - Exposure of phosphatidylserine on the surface of apoptotic cells contributes to their removal in situ by quiet, noninflammatory, and nonimmunogenic mechanisms. In this issue of Immunity, describe a completely new mechanism for this effect that involves a novel zinc finger nuclear factor that inhibits IL-12 production. PMID- 15539148 TI - The acquired immune system: a vantage from beneath. PMID- 15539149 TI - The caveolin-1 binding domain of HIV-1 glycoprotein gp41 is an efficient B cell epitope vaccine candidate against virus infection. AB - Caveolin-1 is a scaffolding protein that organizes and concentrates specific ligands within the caveolae membranes. We identified a conserved caveolin-1 binding motif in the HIV-1 transmembrane envelope glycoprotein gp41 and designed several synthetic peptides, referred to as CBD1, corresponding to the consensus caveolin-1 binding domain in gp41. In rabbits, these peptides elicit the production of antibodies that inhibit infection of primary CD4(+) T lymphocytes by various primary HIV-1 isolates. Interestingly, gp41 exists as a stable complex with caveolin-1 in HIV-infected cells. Anti-CBD1 peptide antibodies, therefore, might be functional by inhibiting the potential interaction of gp41 with caveolin 1. Because of their capacity to elicit antibodies that inhibit the different clades of HIV-1, CBD1-based peptides may represent a novel synthetic universal B cell epitope vaccine candidate for HIV/AIDS. Moreover, such peptides could also have an application as a therapeutic vaccine since CBD1-specific antibodies are rare in HIV-infected individuals from several geographic origins. PMID- 15539150 TI - TRAF2 differentially regulates the canonical and noncanonical pathways of NF kappaB activation in mature B cells. AB - To examine the role of the TNF-R superfamily signaling protein TRAF2 in mature B cell development and NF-kappaB activation, conditionally TRAF2-deficient mice were produced. B cells lacking TRAF2 expression in these mice possessed a selective survival advantage, accumulated in the lymph nodes and splenic marginal zone, were larger in size, and expressed increased levels of CD21/35. These TRAF2 deficient B cells could not proliferate or activate the canonical NF-kappaB pathway in response to CD40 ligation. By contrast, noncanonical NF-kappaB activation was constitutively hyperactive, with TRAF2-deficient B cells exhibiting close to maximal processing of NF-kappaB2 from p100 to p52 and high levels of constitutive p52 and RelB DNA binding activity. These findings establish TRAF2 as a multifunctional regulator of NF-kappaB activation that mediates activation of the canonical pathway but acts as a negative regulator of the noncanonical pathway. This dual functionality explains the contrasting roles of TRAF2 in B cell maturation and activation. PMID- 15539151 TI - Transcriptional suppression of interleukin-12 gene expression following phagocytosis of apoptotic cells. AB - Phagocytosis of apoptotic cells usually results in an anti-inflammatory state with inhibition of proinflammatory cytokines such as IL-12. How apoptotic cell derived signals regulate IL-12 gene expression is not understood. We demonstrate that cell-cell contact with apoptotic cells is sufficient to induce profound inhibition of IL-12 production by activated macrophages. Phosphatidylserine could mimic the inhibitory effect. The inhibition does not involve autocrine or paracrine actions of IL-10 and TGF-beta. We report the identification, purification, and cloning of a novel zinc finger nuclear factor, named GC binding protein (GC-BP), that is induced following phagocytosis of apoptotic cells by macrophages or by treatment with phosphatidylserine. GC-BP selectively inhibits IL-12 p35 gene transcription by binding to its promoter in vitro and in vivo, thus decreasing IL-12 production. Blocking GC-BP by RNA interference restores IL 12 p35 transcription and IL-12 p70 synthesis. Finally, GC-BP itself undergoes functionally significant tyrosine dephosphorylation in response to apoptotic cells. PMID- 15539152 TI - Induction of secondary and tertiary lymphoid structures in the skin. AB - During embryogenesis a developmental program leading to the formation of lymph nodes and Peyer's patches is initiated. We now show that lymph node-like structures as well as tertiary lymphoid structures can ectopically be induced by intradermal injection of newborn lymph node-derived cells. ICAM-1/VCAM-1 expressing stromal organizers, follicular dendritic cells, lymphatic endothelium, and HEVs in these structures are of donor origin, while all hematopoietic cells are host derived. Formation depends on lymphotoxin-expressing donor cells, whereas further organization requires lymphotoxin-expressing recipient cells. While induced secondary lymphoid structures develop a normal cellular architecture, the degree of organization in tertiary structures is correlated to the immune activation status of the host. These results indicate that the cellular and molecular requirements for the establishment of lymph nodes and tertiary structures are remarkably similar and that hyperactivated lymphocytes can fulfill the role of lymphoid tissue inducer cells during inflammatory responses. PMID- 15539153 TI - Clonal selection of helper T cells is determined by an affinity threshold with no further skewing of TCR binding properties. AB - Helper T cell responses that focus the TCR repertoire of responding clones provide experimental access to the mechanisms of clonal selection in vivo. Using TCRbeta chain animals, we directly evaluate the extent of TCRalpha CDR3 diversity and the pMHCII binding attributes of individual antigen-specific Th cells. Here, we demonstrate that dominant clonotypes, as defined by TCR junctional sequence similarities, are surprisingly diverse at the level of pMHCII binding properties, before and after antigen exposure. During an immune response, we can detect and quantify the selective loss of antigen-specific clonotypes that express lower affinity TCR. This affinity threshold selection is followed by the unbiased propagation of preferred clonotypes regardless of TCR-pMHCII half-lives or affinity. Thus, an affinity threshold mechanism discriminates Th clones with TCR of best fit and propagates clonal diversity without promoting autoreactivity. PMID- 15539154 TI - Grb2 and the non-T cell activation linker NTAL constitute a Ca(2+)-regulating signal circuit in B lymphocytes. AB - Activation of the B cell antigen receptor triggers phosphorylation of cytoplasmic and transmembrane adaptor proteins such as SLP-65 and NTAL, respectively. Specific phosphoacceptor sites in SLP-65 serve as docking sites for Ca(2+) mobilizing enzymes Btk and PLC-gamma2. Phosphorylated NTAL recruits the Grb2 linker, but downstream signaling cascades are unclear. We now show that receptor induced tyrosine phosphorylation of NTAL and concomitant Grb2 complex formation critically modulate the Ca(2+) response without affecting SLP-65 and PLC-gamma2 phosphorylation. Grb2 turned out to play a negative regulatory role, which appears to be eliminated upon binding to NTAL. This allows for a sustained release of intracellular Ca(2+) and is mandatory for subsequent entry of Ca(2+) from extracellular sources. Thus, elevation of Ca(2+) is regulated by at least two signaling modules, the B cell-specific Ca(2+) initiation complex comprising SLP-65, Btk, and PLC-gamma2 and the more ubiquitously expressed NTAL/Grb2 complex, which acts as an amplifier by switching off inhibitory elements. PMID- 15539155 TI - SAP regulates T(H)2 differentiation and PKC-theta-mediated activation of NF kappaB1. AB - XLP is caused by mutations affecting SAP, an adaptor that recruits Fyn to SLAM family receptors. SAP-deficient mice recapitulate features of XLP, including increased T cell activation and decreased humoral responses post-infection. SAP deficient T cells also show increased TCR-induced IFN-gamma and decreased T(H)2 cytokine production. We demonstrate that the defect in IL-4 secretion in SAP deficient T cells is independent of increased IFN-gamma production. SAP-deficient cells respond normally to polarizing cytokines, yet show impaired TCR-mediated induction of GATA-3 and IL-4. Examination of TCR signaling revealed normal Ca(2+) mobilization and ERK activation in SAP-deficient cells, but decreased PKC-theta recruitment, Bcl-10 phosphorylation, IkappaB-alpha degradation, and nuclear NF kappaB1/p50 levels. Similar defects were observed in Fyn-deficient cells. SLAM engagement amplified PKC-theta recruitment in wt but not SAP- or Fyn-deficient cells, arguing that a SAP/Fyn-mediated pathway enhances PKC-theta/NF-kappaB1 activation and suggesting a role for this pathway in T(H)2 regulation. PMID- 15539156 TI - Genetic evidence linking SAP, the X-linked lymphoproliferative gene product, to Src-related kinase FynT in T(H)2 cytokine regulation. AB - SAP is an adaptor mutated in X-linked lymphoproliferative disease. It plays a critical role in T helper 2 (T(H)2) cytokine production. This function was suggested to reflect the capacity of SAP to associate with SLAM family receptors and enable tyrosine phosphorylation signaling by these receptors through SAP mediated recruitment of Src-related kinase FynT. Here, we addressed by genetic means the importance of the SAP-FynT interaction in normal T cell functions. By creating a mouse in which the FynT binding site of SAP was inactivated in the germ line (sap(R78A) mouse) and by analyzing mice lacking SAP, FynT or SLAM, evidence was obtained that the SAP-FynT cascade is indeed crucial for normal T(H)2 functions in vitro and in vivo. These data imply that SAP is necessary for T(H)2 cytokine regulation primarily as a result of its capacity to recruit FynT. They also establish a previously unappreciated role for FynT in SAP-dependent T(H)2 cytokine regulation. PMID- 15539157 TI - Silencing T-bet defines a critical role in the differentiation of autoreactive T lymphocytes. AB - As a means of developing therapies that target the pathogenic T cells in multiple sclerosis (MS) without compromising the immune system or eliciting systemic side effects, we investigated the use of T-bet-specific antisense oligonucleotides and small interfering RNAs (siRNA) to silence T-bet expression in autoreactive encephalitogenic T cells and evaluated the biological consequences of this suppression in experimental autoimmune encephalomyelitis, a model for MS. The T bet-specific AS oligonucleotide and siRNA suppressed T-bet expression, IFNgamma production, and STAT1 levels during antigen-specific T cell differentiation. In vitro suppression of T-bet during differentiation of myelin-specific T cells and in vivo administration of a T-bet-specific antisense oligonucleotide or siRNA inhibited disease. T-bet was shown to bind the IFNgamma and STAT1 promoters, but did not regulate the IL-12/STAT4 pathway. Since T-bet regulates IFNgamma production in CD4(+) T cells, but to a lesser extent in most other IFNgamma producing cells, T-bet may be a target for therapeutics for Th1-mediated diseases. PMID- 15539158 TI - Toll-dependent control mechanisms of CD4 T cell activation. AB - Toll-like receptors (TLRs) detect microbial infection and play an essential role in the induction of innate and adaptive immune responses. The mechanisms of TLR mediated control of adaptive immunity are not yet fully understood. Induction of dendritic cell (DC) maturation is essential for activation of naive T cells. Here, we demonstrate that TLR-induced DC maturation and migration to the lymph nodes, in the absence of TLR-induced inflammatory cytokines, are not sufficient for T cell activation in vivo. We show that transient depletion of regulatory T (Tr) cells recovers the primary CD4 T cells response in MyD88-deficient mice, demonstrating that a major mechanism of TLR-mediated activation of T cell responses is the blocking of suppression by regulatory T cells. In addition we show that a TLR-induced signal(s) is required for memory CD4 T cell differentiation, but not for activation of memory T cells. PMID- 15539160 TI - Stable isotope methods for studies of mineral/trace element metabolism. PMID- 15539161 TI - The effect of copper deficiency on rat hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase activity. AB - The effect of copper deficiency on hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase, the key enzyme regulating cholesterol biosynthesis, was investigated in the rat. Male weanling rats were fed semipurified diets containing adequate, marginal, or deficient levels of copper for 6 weeks. Two separate studies were conducted; in the first study, animals were fasted 12 hours prior to analysis and in the second study, animals were fed diets ad libitum. Plasma lipid levels, hepatic cholesterol concentrations, and 3-hydroxy-3-methylglutaryl coenzyme A reductase specific activity, total and active, were determined. Consistent with previous findings, plasma total cholesterol and triglyceride levels were significantly elevated in copper-deficient rats. Copper deficiency resulted in a significant decrease in hepatic total cholesterol levels. Total and active levels of 3-hydroxy-3-methylglutaryl coenzyme A reductase in fed animals were elevated twofold with copper deficiency, with the active form of the enzyme constituting approximately 30% of total activity. 3-Hydroxy-3-methylglutaryl coenzyme A reductase activity in copper-deficient fasted rats was twofold higher than for the fasted adequate animal; however, fasting did result in a 10-fold reduction in hepatic reductase specific activity. These data support the hypothesis that copper deficiency results in a hypercholesterolemic state in the rat associated with increased hepatic cholesterol synthesis. PMID- 15539162 TI - Effect of wheat bran and wheat germ on the intestinal uptake of oleic acid, monoolein, and cholesterol in the rat. AB - The effects of fiber-rich wheat bran and wheat germ on the intestinal absorption of dietary cholesterol, free fatty acids, and monoglycerides were studied. Rats were given a test meal containing [(14)C]oleic acid, [(14)C]monoolein, and [(3)H]cholesterol. After a 1-hour digestion period, wheat bran or wheat germ (10% of meal solids) did not significantly modify the gastric emptying of lipids. No effect of wheat bran was evidenced on the amounts of lipids and cholesterol in the intestinal content or the mucosal segments, whereas wheat germ significantly increased the cholesterol in the small intestine content, decreasing its intestinal absorption. Both fractions only slightly influenced the levels of absorbed lipids and cholesterol in the plasma and liver. In vitro binding measurements showed that the wheat fractions bind only 7% to 15% of both lipids and cholesterol. Results indicate that wheat bran has no direct effect on the mucosal uptake process, whereas wheat germ might decrease the uptake of dietary cholesterol by an as yet unknown mechanism. PMID- 15539163 TI - Interactive effects of dietary silicon, copper, and zinc in the rat. AB - A factorial rat experiment using two dietary concentrations each of copper, zinc, and silicon was conducted to identify areas in which interrelationships involving silicon may exist. The concentrations used were (mg/kg of diet): copper, 1 and 5; zinc, 2 and 12; and silicon, 5 and 270. An antagonism between silicon and zinc, whereby increases in dietary levels of either one resulted in a reduction in blood plasma concentrations of the other, was demonstrated. The depressing effect of silicon on plasma concentrations of zinc and on alkaline phosphatase occurred only in zinc-deficient rats. However, silicon had no effect on growth. Effects on aortic composition, interpreted as beneficial, accompanied increases in the silicon content of copper-deficient diets. Silicon-dependent increases in the chloroform-methanol extractable fraction of aorta closely approximated a similar response to copper. High dietary silicon increased aortic elastin in copper deficient rats when dietary zinc was adequate. The aortic effects of silicon, while mimicking the gross effects of copper, occurred in the absence of any silicon-related changes in blood copper concentrations. Interrelationships of silicon with other elements, particularly copper and zinc, may warrant consideration in future nutritional and metabolic studies. PMID- 15539164 TI - Effect of structured lipid-enriched total parenteral nutrition in rats bearing yoshida sarcoma. AB - The efficacy of structured lipid, a triacylglycerol of medium and long chain fatty acids, as an element of nutritional support therapies in cancer cachexia was investigated. Using the Yoshida sarcoma to induce cachexia, male Sprague Dawley rats (90 g) were injected subcutaneously with tumor cells (n = 17) or sterile saline (n = 16). Seven days later, rats were randomized to two intravenous diets for 3 days at 220 kcal/kg body weight/d, including 2 g nitrogen/kg body weight/d and 39% of total calories as either structured lipid or long chain triglyceride. Nitrogen balance, tumor growth rate, energy metabolism, and plasma albumin and free fatty acid levels were measured, and whole-body protein kinetics and liver, muscle, and tumor fractional protein synthetic rates were evaluated by adding (14)C-leucine to the diet during the last 4 hours of feeding. Nitrogen balance improved (P < .05) in both tumor and control rats receiving structured lipid-enriched total parenteral nutrition, and was also greater in tumor rats compared with controls. There were no differences in tumor growth or protein kinetics between diet groups. Albumin was lower (P < .05) in tumor rats, but significantly higher in both tumor and control rats given structured lipid-enriched total parenteral nutrition. Free fatty acid was significantly higher in tumor rats versus controls. Whole-body protein kinetics were similar among the four groups. Liver weight, liver weight to body weight ratio, and liver protein synthetic rate were higher in tumor rats. Also, liver weight to body weight ratio was lower in tumor and control animals given structured lipid-enriched total parenteral nutrition. Muscle protein synthetic rate was significantly lower in tumor rats, but higher in tumor and control rats given long chain triglyceride-enriched total parenteral nutrition. The nutritional benefits of structured lipid-enriched total parenteral nutrition favor support of host tissue without promoting tumor growth. PMID- 15539165 TI - Effect of dietary tryptophan on plasma and brain tryptophan, brain serotonin, and brain 5-hydroxyindoleacetic acid in rainbow trout. AB - In order to determine the effect of dietary tryptophan level on plasma and brain tryptophan, brain serotonin, and brain 5-hydroxyindoleacetic acid levels, juvenile rainbow trout (Salmo gairdneri) were raised for 16 weeks on semipurified diets containing 0.06%, 0.16%, 0.21%, 0.26%, 0.39%, or 0.59% tryptophan. After 14 weeks, feed intake was depressed in fish fed the diets containing 0.06% or 0.16% tryptophan. No further differences in feed intake were noted between the remaining treatments. In addition, body weight was lower in fish fed diets containing 0.06%, 0.16%, or 0.21% tryptophan compared with fish fed higher levels. After 16 weeks of feeding the test diets, plasma tryptophan levels were found to be directly related to dietary tryptophan levels. Similarly, increased dietary levels of tryptophan resulted in increased brain levels of tryptophan, serotonin, and 5-hydroxyindoleacetic acid. These results demonstrate that in rainbow trout, as in mammals, altered dietary levels of tryptophan result in alterations in plasma and brain tryptophan, brain serotonin, and brain 5 hydroxyindoleacetic acid. PMID- 15539166 TI - Choline: determination using gas chromatography/mass spectrometry. PMID- 15539167 TI - Aromatic amino acids and monoamine synthesis in the central nervous system: influence of the diet. PMID- 15539168 TI - Suppression of avian hepatic cholesterogenesis by dietary ginseng. AB - The effect of the ginseng root powder on avian hepatic cholesterol biosynthesis and serum lipoprotein cholesterol levels were examined. Lohman strain broiler females were fed for 4 weeks a corn-based diet (control) or an experimental diet in which 0.25% Korean ginseng was incorporated (treatment). B.-hydroxy-B methylglutaryl-CoA) HMG-CoA reductase activity was significantly lower (P < 0.01) in the treatment group (47% of control activity). Ginseng treatment affected a lowering of the serum total cholesterol level (83% of control, (P < 0.05) and of serum low density lipoprotein cholesterol level (77% of control, P < 0.05). The mechanism of the hypocholesterolemic action of ginseng involves the suppression of cholesterol biosynthesis. PMID- 15539169 TI - Effect of diabetes on the rat hepatic glucose-6-phosphatase system in endoplasmic reticulum subfractions. AB - Diabetes-induced alterations in the activities of the components of the glucose-6 phosphatase system (i.e., the enzyme, the glucose-6-P translocase (T(1)), and the phosphate translocase (T(2)) were examined in smooth and rough subfractions of hepatic endoplasmic reticulum from streptozotocin-injected rats. A significant effect of diabetes on the maximal velocity of glucose-6-P hydrolysis by the enzyme was present in both endoplasmic reticulum subfractions (3.1-fold increase in rough endoplasmic reticulum; 3.8-fold increase in smooth endoplasmic reticulum). Based on latency values, diabetes did not result in a proportional increase in capacity of T(1) or T(2). In contrast to the control condition, the relationship between transport capacity and hydrolytic capacity was not significantly different in the two subfractions from diabetic animals. Elucidation of the effects of diabetes on the components of the glucose-6 phosphatase system associated with smooth and rough endoplasmic reticulum membranes enhances our understanding of the hepatic contribution to diabetic hyperglycemia. PMID- 15539170 TI - Kinetics of tissue RRR-alpha-tocopherol depletion and repletion. Effect of cold exposure. AB - Vitamin E was estimated in plasma and tissues of rats kept for three months on a low vitamin E diet or a high vitamin E diet. Some of the animals from each group were switched to the opposite diet, and the kinetics of uptake and depletion of vitamin E were followed 3, 8, and 15 days after the diet change. Some rats were also submitted to cold exposure (6 degrees C) for three days. During repletion plasma, red blood cells, liver, spleen, and adrenal gland were the only tissues that responded rapidly to the diet change; after three days, their vitamin E levels corresponded to that of the new diet. Heart, brain, lung, muscle, and thymus were slow in reacting to diet change. Fifteen days after the change in diet, white adipose tissue did not respond. The rate of repletion for all tissues was more rapid than the rate of depletion, but liver was the only tissue that after three days had vitamin E levels corresponding to the low-vitamin diet. Cold exposure for three days did not produce any significant change in the vitamin E content of any tissue, indicating that despite high oxygen consumption by the animal, vitamin E was not consumed or mobilized. PMID- 15539171 TI - Effects of choline deficiency and methotrexate treatment upon rat liver. AB - Choline deficiency and treatment with methotrexate (MTX) both are associated with fatty infiltration of the liver. Choline, methionine, and folate metabolism are interrelated and converge at the regeneration of methionine from homocysteine. MTX perturbs folate metabolism, and it is possible that it also influences choline metabolism. We fed rats a choline deficient diet for 2 weeks and/or treated them with methotrexate (MTX; 0.1 mg/kg daily). Choline deficiency lowered hepatic concentrations of choline (to 43% control), phosphocholine (PCho; to 18% control), glycerophosphocholine (GroPCho; to 46% control), betaine (to 30% control), phosphatidylcholine (PtdCho; to 62% control), methionine (to 80% control), and S-adenosylmethionine (AdoMet; to 57% control), while S adenosylhomocysteine (AdoHcy) and triacylglycerol concentrations increased (to 126% and 319% control, respectively). MTX treatment alone lowered hepatic concentrations of PCho (to 48% control), GroPCho (to 69% control), betaine (to 55% control), and AdoMet (to 75% control). The addition of MTX treatment to choline deficiency resulted in a larger decrease in AdoMet concentrations (to 75% control) and larger increases in AdoHcy and triacylglycerol concentrations (to 150% and 500% control, respectively) than was observed in choline deficiency alone. Livers from MTX-treated animals used radiolabeled choline to make the same metabolites as did livers from controls (most of the label was converted to PCho and betaine). In choline deficient animals, most of the labeled choline was converted to PtdCho. Therefore, MTX depleted hepatic PCho, GroPCho, and betaine by a mechanism that was different from that of choline deficiency. MTX increased the extent of fatty infiltration of the liver in choline deficient rats, and choline deficiency and MTX treatment damaged hepatocytes as measured by leakage of alanine aminotransferase activity. Our data are consistent with the hypothesis that the fatty infiltration of the liver associated with MTX treatment occurs because of a disturbance in choline metabolism. PMID- 15539172 TI - Effect of various chloride salts on the utilization of phosphorus, calcium, and magnesium. AB - Only part of the effect of dietary protein on urinary calcium excretion can be ascribed to sulfur amino acids. We hypothesized that chloride, another factor often associated with isolated proteins, and another amino acid, lysine, affect utilization of calcium. The effects of supplemental dietary chloride, inorganic or organic, on calcium, phosphorus, and magnesium utilization were studied in two rat studies. Weanling Sprague-Dawley rats were fed semi-purified diets that contained moderate (1.8 mg Cl/g diet) or supplemental (15.5 mg Cl/g diet) chloride as sodium chloride, potassium chloride, or lysine monohydrochloride with or without calcium carbonate for 56 or 119 days. Rats fed supplemental sodium chloride or potassium chloride had higher urinary phosphorus excretion, more efficient phosphorus absorption, but unchanged tissue phosphorus levels after 7 and 16 weeks of dietary treatment as compared to rats fed moderate chloride. Rats fed supplemental sodium chloride or potassium chloride excreted more calcium in urine at 7 weeks and absorbed calcium less efficiently at 16 weeks. Tissue calcium concentrations were unaffected, but total tibia magnesium and plasma magnesium concentrations were lower in rats fed supplemental sodium chloride or potassium chloride than those fed moderate chloride. Lysine chloride with or without additional calcium elevated urinary calcium excretion even more than sodium chloride and potassium chloride ingestion. Rats fed lysine chloride with supplemental calcium had smaller apparent absorption and urinary losses of phosphorus and magnesium after 16 weeks and lower tibia and plasma magnesium concentrations than rats fed lysine chloride. PMID- 15539173 TI - Analysis of lipids by high performance liquid chromatography. Part II: phospholipids. PMID- 15539175 TI - Dietary n-9, n-6, and n-3 fatty acids modify linoleic acid more than arachidonic acid levels in plasma and platelet lipids and minimally affect platelet thromboxane formation in the rabbit. AB - We have studied the effects of semisynthetic diets containing 5% by weight (12% of the energy) of either olive oil (70% oleic acid, OA) or corn oil (58% linoleic acid), or fish oil (Max EPA, containing about 30% eicosapentaenoic, EPA C 20:5 n 3, plus docosahexaenoic, DHA C 22:6 n-3, acids, and less than 2% linoleic acid), fed to male rabbits for a period of five weeks, on plasma and platelet fatty acids and platelet thromboxane formation. Aim of the study was to quantitate the absolute changes of n-6 and n-3 fatty acid levels in plasma and platelet lipid pools after dietary manipulations and to correlate the effects on eicosanoid precursor fatty acids with those on platelet thromboxane formation. The major differences were found when comparing the group fed fish oil and depleted linoleic acid vs the other groups. The accumulation of n-3 fatty acids in various lipid classes was associated with modifications in the distribution of linoleic acid and arachidonic acid in different lipid pools. In platelets maximal incorporation of n-3 fatty acids occurred in phosphatidyl ethanolamine, which also participated in most of the total arachidonic acid reduction occurring in platelets, and linoleic acid, more than archidonic acid, was replaced by n-3 fatty acids in various phospholipids. The archidonic acid content of phosphatidyl choline was unaffected and that of phosphatidyl inositol only marginally reduced. Thromboxane formation by thrombin stimulated platelets did not differ among the three groups, and this may be related to the minimal changes of arachidonic acid in phosphatidyl choline and phosphatidyl inositol. PMID- 15539176 TI - Relationship of milk iron and the changing concentration of mammary tissue transferrin receptors during the course of lactation. AB - The concentration of iron in all mammalian milks falls during lactation while the infant's iron requirement increases. Little is known, however, about the entry of iron into milk. Recently, transferrin receptors have been identified on lactating rat mammary plasma membranes, which may regulate iron entry into mammary tissue, potentiating its availability for subsequent transport into milk. This study was conducted to determine what relationship exists between the declining concentration of milk iron and the transferrin receptor concentration during various stages of lactation. Minimal transferrin receptors were detected in nulliparous rats. Total mammary transferrin receptor content increased during early and mid-lactation while milk iron concentration decreased. The continued appearance of high levels of transferrin receptors throughout lactation, without a concomitant increase in milk iron concentration, suggests a need for iron for functions other than cellular growth or secretion into milk to meet infant needs. PMID- 15539177 TI - Sequential changes in alanine metabolism following partial hepatectomy in the rat. AB - After partial hepatectomy, the liver undergoes an array of metabolic changes until regeneration is complete. Since carbons derived from alanine can be incorporated into most metabolic pools, we studied the metabolism of (14)C labeled alanine during the early phase of regeneration. Sham operated (controls) and partially hepatectomized rats weighing about 200 g each were injected intraperitoneally with 1-[U-(14)C]alanine at 9, 18, and 36 hours after surgery. The animals were killed 2 hours after injection. Compared to the controls, alanine oxidation was markedly depressed (P < 0.05) in the 9- and 18-hour groups, but was restored in the 36-hour group. The specific activity of plasma glucose and hepatic glycogen was elevated 9 and 18 hours after partial hepatectomy. There was a corresponding increase in the activities of fructose-1,6-diphosphatase and phosphoenolpyruvate carboxykinase. Hepatic protein specific activity increased by 30, 74, and 120%, respectively 9, 18, and 36 hours after partial hepatectomy. Hepatic fatty acids followed a similar pattern. In a separate set of experiments, the distribution of radioactivity in glutamic acid was measured. The results showed that alanine carbons enter the citric acid cycle primarily via the acetyl CoA pathway in the controls, but via the oxaloacetate pathway in partially hepatectomized rats. The results demonstrate significant changes in the activities of metabolic pathways of alanine in the early phase of hepatic regeneration. PMID- 15539178 TI - Effects of dietary calcium on the metabolism of trace elements in male and female rats. AB - The effect of dietary calcium on the metabolism of iron, zinc, copper, and manganese in male and female rats was investigated. For 3 or 6 weeks the rats were fed three diets containing: (1) 0.26, (2) 0.52, or (3) 2.08% Ca. The apparent absorption of iron was depressed by the high calcium diet, and manganese absorption was highest in the low calcium groups. Generally there was a decrease in the absorption of minerals from 3 to 6 weeks. With an increase in the dietary calcium the absorption of Ca and P decreased. The liver iron concentration in the females fed diet 3 decreased from about 600 to 200 microg/g dry weight. The high calcium intake also caused a slight increase in the heart calcium levels in both sexes. However, diet 3 prevented kidney calcification in the female rats at 6 weeks and this was attributed to a dramatic decrease in the urinary phosphorus, although the calcium had increased about 40 times. In males, on the other hand, the high calcium diet caused some kidney calcification. PMID- 15539179 TI - Isolation, characterization, and developmental expression of pig intestinal fatty acid-binding proteins. AB - The goal of this study was to characterize and quantify intestinal fatty acid binding proteins of the pig. Small intestinal mucosa from 13-19 kg pigs was homogenized and centrifuged to obtain cytosol. Isolation of fatty acid-binding proteins from delipidated cytosol was achieved using molecular sieve, oleic acid affinity, and ion exchange chromatography. Fatty acid-binding protein isolation was monitored using a fatty-acid binding assay in conjunction with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. Antisera to rat liver-fatty acid-binding protein cross reacted with an isolated intestinal fatty acid-binding protein of Mr = 13,000, whereas antisera to rat intestine-fatty acid binding protein was not cross reactive with isolated pig intestinal proteins. These experiments identify a pig intestinal fatty acid-binding protein that exhibits strong immunochemical similarity to rat liver-fatty acid-binding protein. Cytosol prepared from intestinal mucosa of pigs at -4, 2, 4, 7, 15, 22, 28, and 35 d of age was assayed for fatty acid-binding protein activity. Preweaning fatty acid-binding protein activity in cytosol was maximal at 7 days of age when expressed as total jejunal fatty acid binding per kilogram bodyweight, intestinal or mucosal weight or milligram total protein. After weaning (21 d), fatty acid-binding protein activities declined to 28 days, but increased again by 35 days. Total soluble fatty acid-binding protein activity in pig intestine is regulated during postnatal development and this may account in part for the altered intestinal absorption of lipids observed in young pigs at weaning. PMID- 15539180 TI - Implementation of SI units for clinical laboratory data: style specifications and conversion tables. PMID- 15539181 TI - Hepatic mechanisms for clearance and detoxification of bacterial endotoxins. PMID- 15539182 TI - Effects of iron deficiency upon the antibody response to influenza virus in rats. AB - The effects of severe and moderate iron deficiency upon the antibody response to influenza virus were investigated in rats. Three groups of weanling male Wistar rats were fed one of two iron-deficient diets (5 mg and 15 mg iron/kg diet) or a normal iron-containing diet (35 mg iron/kg diet). A group of individually pair fed rats was introduced with the low iron-consuming rats. The effects of the diets upon various iron status parameters were followed during the 4th, 5th, 6th, and 7th week of diet. After 4 weeks of feeding different diets, an intraperitoneal injection of inactivated influenza virus A/New Jersey/76 was performed and a recall injection was done at 5 weeks. Primary and secondary antibody responses were assayed. Rats were sacrificed at 7 weeks of diet. After 4 weeks of feeding different diets, the rats fed the 5 mg iron/kg diet were severely anemic and rats fed 15 mg iron/kg diet were moderately iron-deficient, as shown by their iron status parameters. Growth was delayed in anemic and matched pair-fed rats. A primary antibody response was almost nonexistent in all groups. Secondary antibody titers were significantly weaker in anemic rats than in ad libitum controls, but were not different from those of pair-fed rats. This response was similar in moderately iron-deficient, ad libitum, and pair-fed rats. These results show that antibody synthesis in response to the influenza virus vaccine is preserved in moderate iron deficiency but is reduced in severe anemia. The reduction in energy consumption associated with severe iron deficiency in the rat could play a part in the altered humoral response. PMID- 15539183 TI - Effect of thiamin deficiency on energy metabolites in the turkey. AB - The effects of thiamin deficiency on selected energy-related metabolites was investigated. A basal diet (B) was formulated to be 11% of NRC recommended level of 2 mg/kg of thiamin. Thiamin was added to this basal diet to generate the control diet (C). Twenty one-week-old female turkeys were fed either the B or C diet. On days four and five of the experiment, food intake was decreased significantly in B fed turkeys (P < 0.05). Plasma and brain samples were collected at this time. Brains were dissected and analyzed for ATP, ADP, uric acid, free fatty acids, glucose, and GABA. Adenosine triphosphate and the ATP/ADP ratio were decreased in the hindbrain (medulla-pons area) of thiamin deficient birds (P < 0.01). Uric acid was increased (P < 0.001) and free fatty acids were decreased (P < 0.0005) in the plasma of thiamin deficient birds. Based on the data, changes in ATP and ATP/ADP levels may be related to the anorectic behavior exhibited by the thiamin deficient bird. PMID- 15539184 TI - Effects of skim milk, skim milk yogurt, orotic acid, and uric acid on lipid metabolism in rats. AB - The effects of feeding two milk products (skim milk and skim milk yogurt) and two proposed hypocholesterolemic factors (orotic acid and uric acid) on serum cholesterol (HDL, LDL, total, HDL/Total and HDL/LDL), liver lipids (total liver lipids and liver cholesterol), and aortal cholesterol were studied. Ten groups, of nine rats each, were fed isocaloric Chow-based diets containing water, 45% skim milk (SM), 45% skim milk yogurt (SMY), and 0.0025% orotic acid (OA) or 0.001% uric acid (UA), without or with cholesterol. The SM diet (with cholesterol) resulted not only in lower total cholesterol (P < 0.10), LDL cholesterol (P < 0.05), aortal cholesterol (P < 0.01), and liver cholesterol (P < 0.10), but also in increased HDL (P < 0.05) and HDL/LDL (P < 0.10) cholesterol ratio. The SMY diet, on the other hand, resulted in lowered total serum cholesterol (P < 0.05) and aortal cholesterol (P < 0.01) and in higher LDL (P < 0.05) cholesterol. The hypocholesterolemic effects were more marked for SM than for SMY. Addition of OA and UA to diets increased serum cholesterol, LDL cholesterol, and total liver lipids; the OA diet also increased liver cholesterol. Neither OA nor UA alone was the factor responsible for the hypocholesterolemic effects seen with SM and SMY feeding. PMID- 15539185 TI - Non-everted oxygenated rat intestinal segments as a measure of neutral detergent fiber effects on iron absorption. AB - Iron absorption in the presence of varying amounts and sizes of dietary fiber was measured. A method using non-everted rat intestinal segments perfused in oxygen was refined. Neutral detergent fiber (NDF), a component of dietary fiber, was extracted from cooked pinto bean (Phaseolus vulgaris). The NDF did not affect iron absorption in intestinal segments from iron replete rats. However, 4 and 6 mg of NDF/ml significantly decreased iron absorption in the intestinal segments from anemic rats. NDF with a smaller particle size of 0.125 mm increased iron absorption relative to that absorbed with 0.180 mm particles. Histological examination validated using non-everted intestinal segments perfused with oxygen as a method for studying dietary effects on iron absorption. Segments which are not everted are less prone to damage. Perfusion with oxygen maintained metabolic activity in the tissue during the experiment. PMID- 15539186 TI - Effect of oral methionine on blood lipid peroxidation and antioxidants in alloxan induced diabetic rats. AB - Supplementation of thiol compounds has been suggested to protect against the toxic effects of reduced oxygen species by contributing to the thiol pool of the cell. The present study was designed to determine whether supplementation of methionine in the diet of diabetic animals protected against the oxidative stress in diabetic pathology. Oral methionine was administered at a dosage of 330 mg/100 g feed to diabetic rats. The effect was compared with the effect of insulin administration. Levels of lipid peroxides were measured in plasma, erythrocytes, and erythrocyte membrane. Anti-oxidants were measured in plasma. Diabetic condition was associated with increased lipid peroxidation and depletion in antioxidant levels. Although methionine did not affect the level of blood glucose and some of the antioxidants, it lowered the lipid peroxide content in blood. Erythrocyte lipid peroxidation activity was unaffected by methionine treatment. Administration of insulin lowered both plasma and erythrocyte lipid peroxide levels. PMID- 15539187 TI - B6 vitamers: cation exchange HPLC. PMID- 15539188 TI - Dietary fatty acids and membrane protein function. AB - In recent years, there has been growing public awareness of the potential health benefits of dietary fatty acids, and of the distinction between the effects of the omega6 and omega3 polyunsaturated fatty acids that are concentrated in vegetable and fish oils, respectively. A part of the biologic effectiveness of the two families of polyunsaturated fatty acids resides in their relative roles as precursors of the eicosanoids. However, we are also beginning to appreciate that as the major components of the hydrophobic core of the membrane bilayer, they can interact with and directly influence the functioning of select integral membrane proteins. Among the most important of these are the enzymes, receptors, and ion channels that are situated in the plasma membrane of the cell, since they carry out the communication and homeostatic processes that are necessary for normal cell function. This review examines current information regarding the effects of diet-induced changes in plasma membrane fatty acid composition on several specific enzymes (adenylate cyclase, 5'-nucleotidase, Na(+)/K(+)-ATPase) and cell-surface receptors (opiate, adrenergic, insulin). Dietary manipulation studies have demonstrated a sensitivity of each to a fatty acid environment that is variably dependent on the nature of the fatty acid(s) and/or source of the membrane. The molecular mechanisms appear to involve fatty acid-dependent effects on protein conformation, on the "fluidity" and/or thickness of the membrane, or on protein synthesis. Together, the results of these studies reinforce the concept that dietary fats have the potential to regulate physiologic function and to further our understanding of how this occurs at a membrane level. PMID- 15539189 TI - 1,25(OH)2D3 increases calcium and phosphatidylinositol metabolism in differentiating cultured human keratinocytes. AB - The effect of 1,25(OH)(2)D(3) on the intracellular calcium, (Ca(+2))i, in both cultured human keratinocytes and in cultured human dermal fibroblasts was investigated. When the intracellular calcium (Ca(+2))i in cultured human keratinocytes, grown in a serum-free medium containing 1.8 mM calcium, was measured by the fluorescent calcium-indicator, Furu-2, the (Ca(+2)i increased 154%, 202%, and 409% over the control value after incubation with 1,25(OH)(2)D(3) at 10(-10) m, 10(-8) m, and 10(-6) m, respectively. This response was immediate (15 seconds), specific (no effect with either 25(OH)D(3) at 10(-8) m or vitamin D(3) at 10(-8) m), and occurred with or without EGTA in the medium. In contrast, 1,25(OH)(2)D(3) did not increase the (Ca(2+))i in either cultured human keratinocytes that were grown in low calcium (0.05 mm), serum-free medium or in cultured human dermal fibroblasts that were grown in medium containing 0.05 mm calcium and 1% serum. The effect of 1,25(OH)(2)D(3) on the the turnover of phosphatidylinositol was investigated as a possible cause for the observed increase in (Ca(+2)i. Cultured human keratinocytes that were incubated with (3)H inositol demonstrated a 50 % +/- 10% increase in the triphosphated, plasma membrane-bound metabolite of phosphatidylinositol, PIP(2), by 15 seconds, followed by a rapid decrease at 30 seconds, then a return toward basal levels by 1 minute. Lysophosphatidylinositol, which results from the sn-2 deacylation of phosphatidylinositol by phospholipase A(2), decreased 20% +/- 8% within 30 seconds, then increased to 200% +/- 10% of the control value by 5 minutes. The accumulation of IP(3) was increased 50% to 100% above the control value within 30 seconds and this increase was substained during the 5-minute incubation period. Stimulation of phosphatidylinositol turnover by 1,25(OH)(2)D(3) was not detected in either cultured human keratinocytes that were grown in serum-free, low calcium medium or in cultured human dermal fibroblasts that were grown in 1% serum. PMID- 15539190 TI - Formation of trimethylamine from dietary choline by Streptococcus sanguis I, which colonizes the mouth. AB - Choline is a component of the normal diet, and when humans ingest large amounts they excrete trimethylamine (which can impart a fishy body odor). In the presence of nitrite, trimethylamine can be converted to dimethylnitrosamine, a potent carcinogen. Bacteria in the large intestine metabolize choline to form trimethylamine. We determined that a bacterium normally present in the oral cavity also has this capacity. Mixed bacterial flora cultured from dental plaque and saliva converted choline to trimethylamine. The only organism with trimethylamine-forming capability isolated from these mixed cultures was identified as Streptococcus sanguis I (a facultative anaerobe). The other products formed when choline was cleaved were ethanol and acetate. The formation of trimethylamine by S. sanguis I was enzyme-mediated. Activity was destroyed by heating at 100 degrees C, and obeyed Michaelis-Menten kinetics (K(apparent) for choline = 184 +/- 58 microM; V(max apparent) = 1.7 +/- 0.1 micromol/mg protein/h). Activity was maximal at pH 7.5 to 8.5, was membrane-bound, and required a divalent metal cation (cobalt or iron). More trimethylamine was produced by bacteria incubated under a nitrogen than under an aerobic atmosphere. Activity was inhibited by deanol, betaine aldehyde, hemicholinium-3, iodoacetate, semicarbazide, and 2,4-dinitrophenol, and was enhanced by sulfhydryl-reducing agents (glutathione, 2-mercaptoethanol, DL-dithiothreitol) and sodium bisulfite. The enzyme activity that we describe in S. sanguis I is similar to that previously described in the anaerobic bacteria isolated from intestinal flora. PMID- 15539191 TI - The effect of vitamin E on lipid peroxidation in the copper-deficient rat. AB - The present study was designed to determine whether the supplementation of vitamin E in the copper-deficient diet would ameliorate the severity of copper deficiency in fructose-fed rats. Lipid peroxidation was measured in the livers and hearts of rats fed a copper-deficient diet (0.6 microg Cu/g) containing 62% fructose with adequate vitamin E (0.1 g/kg diet) or supplemented with vitamin E (1.0 g/kg diet). Hepatic lipid peroxidation was significantly reduced by vitamin E supplementation compared with the unsupplemented adequate rats. In contrast, myocardial lipid peroxidation was unaffected by the level of vitamin E. Regardless of vitamin E supplementation, all copper-deficient rats exhibited severe signs of copper deficiency, and some of the vitamin E-supplemented rats died of this deficiency. These findings suggest that although vitamin E provided protection against peroxidation in the liver, it did not protect the animals against the severity of copper deficiency induced by fructose consumption. PMID- 15539192 TI - Effect of acute selenium restriction on whole body endogenous selenium and the selenite-exchangeable metabolic pool in the adult rat. AB - The time course of changes in whole body endogenous selenium (Se(end)) was investigated during a short-term (7-day) selenium restriction study in the adult rat. The method of continuous feeding with a stable isotope of selenium was used to permit normal intake of selenium while distinguishing between the dietary and endogenous components of body selenium. Additionally, the effect of short-term selenium restriction on the time course of the selenite-exchangeable metabolic pool (Se-EMP) was investigated. Two groups of adult male rats were intubated with the in vivo stable isotope (74)SeO(3)(2-), then fed a Torula yeast diet (selenium <0.02 microg/g) and either deionized water (-Se group) or deionized water containing selenium as (76)SeO(3)(2-) (0.1 microg selenium/ml) (+Se group). Three animals from each group were killed at 24-hour intervals. Whole body Se(end) and the estimated size of Se-EMP (W(Se-EMP)) were determined using hydride generation inductively coupled plasma mass spectrometry for isotopic measurements. Whole body Se(end) decreased linearly in the +Se group (Se degrees (end): 54.4 microg; Se(end) at 3 days: 49.3 +/- 2.1; Se(end) at 7 days: 45.2 +/- 2.2). The decrease was exponential for the -Se group (Se degrees (end): 54.4 microg; Se(end) at 3 days: 42.9 +/- 0.3; Se(end) at 7 days: 42.2 +/- 0.7). The value of W(Se-EMP,pl) (microg) was 19.8 +/- 0.6 at 1 day and 19.7 +/- 1.0 at 7 days for the +Se group. The corresponding values for the -Se group were 15.7 +/- 1.5 and 18.8 +/- 0.4. All respective values of W(Se-EMP,pl) for the -Se group were significantly smaller than for the +Se group (P < 0.05), with the exception of values at days 6 and 7. The value of W(Se-EMP,urine) (microg) was 2.1 +/- 0.2 at 1 day, increasing rapidly to 23.5 +/- 1.5 at 7 days for the +Se group. The corresponding values for the -Se group were 3.0 and 23.1. PMID- 15539193 TI - Effects of growth, food intake, and dietary zinc on diadenosine tetraphosphate concentrations in rats. AB - The nucleotide diadenosine tetraphosphate has been suggested to function as a signal molecule for the initiation of DNA replication. Previous studies have indicated that diadenosine tetraphosphate is synthesized by certain aminoacyl tRNA synthetases and that diversion of AMP from the amino acid-enzyme complex to ATP to form diadenosine tetraphosphate is facilitated by zinc ions. The growth retardation of zinc-deficient rats is associated with specific reduction in DNA replication and also with a potentially growth-limiting decrease in food intake. The possibility has been investigated that in zinc-deficient rats, lack of Zn(2+) restricts diadenosine tetraphosphate synthesis, resulting in a failure to synthesize DNA and in a reduction in growth. The results indicate that the depressed growth potential caused by the reduction in food intake associated with the deficiency was sufficient to lower diadenosine tetraphosphate concentrations significantly in the liver and spleen. However, there was no indication of a specific effect of zinc deficiency on diadenosine tetraphosphate values. PMID- 15539194 TI - Carnitine: determination of total carnitine using a radioenzymatic assay. PMID- 15539195 TI - Dietary unsaturated fatty acids: interactions and possible needs in relation to eicosanoid synthesis. AB - In addition to providing energy and essential fatty acids, dietary fatty acids can affect numerous biochemical and physiologic reactions related to secretory, cardiovascular, and immune functions. The major dietary unsaturated fatty acid, linoleic acid, affects tissue arachidonic acid and can influence eicosanoid mediated reactions. Chronic, excess, or imbalanced eicosanoid synthesis may be conductive to excessive inflammation, thrombotic tendencies, atherosclerosis, and immune suppression. Dietary n-3 polyunsaturated fatty acids (PUFAs) may ameliorate eicosanoid-related phenomena by reducing tissue arachidonic acid and by inhibiting eicosanoid synthesis. This review summarizes information concerning the metabolism of unsaturated fatty acids, with emphasis on tissue arachidonic acid levels and eicosanoids, and discusses the need for data concerning the appropriate intake of dietary n-6 and n-3 PUFAs to modulate arachidonic acid and eicosanoid synthesis and to minimize possible adverse reactions. PMID- 15539196 TI - Glutathione peroxidase activities during selenium depletion of adult female rats and during selenium repletion of their offspring. AB - The main purpose of the present investigation was to produce young rats with severe selenium deficiency, but with no clinical signs of this deficiency, and to examine their liver and red blood cell (RBC) glutathione peroxidase activities during selenium repletion. To achieve this goal, female breeders were fed a selenium-deficient diet beginning 2 weeks before mating. The liver glutathione peroxidase activity of the dams was significantly lower than the activity of comparable nonpregnant females after 5 and 10 weeks of selenium depletion. This difference arose exclusively during the period of pregnancy. In contrast, the RBC glutathione peroxidase activity was significantly increased during this period. Only traces of liver enzyme activity were found in the offspring, and the RBC enzyme activity was only 2% of that of the selenium-repleted controls. Body weight was retarded in the male offspring. However, no severe signs of clinical selenium deficiency were observed. The glutathione peroxidase activity in the liver and RBCs of the offspring was determined after 0, 2, 4, 7, 14, and approximately 40 days of selenium repletion. The liver enzyme activity increased faster in females than in males, while the opposite was found for the RBCs. After 14 days of selenium repletion, the glutathione peroxidase activity of the liver was essentially restored, and the RBC enzyme activity was about half that of the control values. This type of rat may prove useful in studies in which young selenium-deficient rats are preferable, as well as in studies of selenium functions that might not be directly related to the role of selenium in glutathione peroxidase. PMID- 15539197 TI - Effect of dietary copper deficiency on the distribution of dopamine and norepinephrine in mice and rats. AB - Dietary copper deficiency was produced in Swiss albino mice and Sprague Dawley rats to determine the organ specificity of alterations in norepinephrine (NE) and dopamine (DA) concentrations and the relationship with organ copper levels. A 5 week dietary treatment was used, which started 1 week after birth for mice, initially via dams, and 3 weeks after birth for rats. Mice offspring (6 weeks of age) and rats (8 weeks of age) maintained on a copper-deficient (-Cu) treatment were compared with copper-adequate (+Cu) controls. Compared with +Cu animals, -Cu mice and rats were anemic and had low (<1% of +Cu) ceruloplasmin activities but normal body weights. The -Cu mice had organ copper concentrations ranging between 30% and 65% of +Cu values for eight organs studied, with the thymus being the least depleted. For -Cu rats, the range was 15% to 65%. Significant reductions in NE concentration were observed in the heart, pancreas, and spleen of -Cu mice. Elevated DA levels were observed in all organs except the brain. For -Cu rats, the NE level was lower in the heart and the DA level was higher in both the heart and spleen compared with +Cu rats. Dopamine elevation in the heart and spleen for both -Cu mice and rats was four- and fivefold higher, respectively. Adrenal catecholamine levels were only slightly changed by copper deficiency in mice or rats. Urinary levels of both NE and DA were higher in -Cu rats and mice. Plasma and heart tyrosine levels were not altered in -Cu mice. Elevated DA in -Cu rodents may be due to limiting dopamine-beta-monooxygenase. Higher urinary NE and lower organ NE may be due to a combination of decreased synthesis and enhanced turnover. The magnitude of decreased organ copper was not predictive of altered catecholamine pool size. PMID- 15539198 TI - The effect of fasting time on plasma total cholesterol concentration in Mongolian gerbils. AB - This study investigated the effect of length of fasting time on plasma total cholesterol response of male Mongolian gerbils (Meriones unguiculatus). Plasma cholesterol levels from fed and fasted gerbils were also compared with those reported for humans under similar metabolic states. Plasma total cholesterol response showed a significant quadratic relationship with time over a 15-hour period. Between 6 and 9 hours of fasting (the time during which plasma triglyceride concentration became relatively constant), the average plasma total cholesterol concentration was 178 mg/dl, compared with a zero hour (fed) cholesterol level of 265 mg/dl. The difference in plasma cholesterol levels observed in fed and fasted gerbils is unlike what has been reported for humans. Results from most human studies show no differences in plasma total cholesterol concentrations for fed and fasted subjects. Failure to consider species differences in metabolic responses may have implications when results from animal experiments are extrapolated to humans. PMID- 15539199 TI - Effect of changing the type of dietary carbohydrate or copper level of copper deficient, fructose-fed rats on tissue sorbitol concentrations. AB - This study was designed to examine the relationship between the fructose-copper interaction and tissue sorbitol concentrations. Weanling male rats were provided with a diet which contained 62.7% fructose and 0.6 microg copper/g (F-Cu) for 4 weeks. At this time, rats were changed to either a fructose diet which contained 6.0 microg copper/g or to a starch diet with or without copper for 2 weeks. When compared with the other dietary groups, it was found that rats fed the F-Cu diet grew poorly; had altered relative liver, pancreatic, heart, and kidney sizes; were anemic; and had higher tissue concentrations of pancreatic and heart glucose, liver, pancreatic, heart, and kidney fructose, and liver, pancreatic, and kidney sorbitol. When rats were changed from the F-Cu diet to one containing copper or to a starch diet with or without copper, weight gain, relative liver, pancreatic and heart sizes, and hematocrit improved significantly. In general, there was a reduction in pancreatic and heart glucose; liver, pancreatic, heart, and kidney fructose; and pancreatic and kidney sorbitol concentrations when rats were changed from the F-Cu diet to any of the other diets. We conclude that the fructose-copper interaction may have a common biochemical basis related to the metabolism of glucose, fructose, and sorbitol. PMID- 15539200 TI - Insulin secretion and glucose uptake in hypomagnesemic sheep fed a low magnesium, high potassium diet. AB - Hyperglycemic and euglycemic clamp experiments were conducted to evaluate insulin secretion and glucose uptake in the hypomagnesemic sheep fed a low magnesium (Mg), high potassium (K) diet. Five mature sheep were fed a semipurified diet containing 0.24% Mg and 0.56% K (control diet) and five were fed 0.04% Mg and 3.78% K (low Mg/high K diet) for at least 2 weeks. In the hyperglycemic clamp experiment, plasma glucose concentrations were raised and maintained at a hyperglycemic steady-state (approximately 130 mg/100 ml) by variable rates of glucose infusion during the experimental period (120 minutes). The insulin response in the sheep fed the low Mg/high K diet (31.0 microU/ml) were significantly (P < 0.01) lower than those (111.7 microU/ml) of the sheep fed the control diet. In the euglycemic clamp experiment, insulin was infused at rates of 5, 10, 15, or 20 mU/kg/min, each followed by variable rates of glucose infusion to maintain a euglycemic steady-state (basal fasting levels). Hypomagnesemic sheep fed the low Mg/high K diet had significantly (P < 0.01) lower mean glucose disposal (3.72 mg/kg/min) across the insulin infusion rates compared with those of the sheep fed the control diet (5.37 mg/kg/min). These results suggest that glucose-induced insulin secretion and insulin-induced glucose uptake would be depressed in hypomagnesemic sheep and are caused by feeding the low Mg/high K diet. PMID- 15539201 TI - Adenosine triphosphate: enzymatic spectrophotometric determination. PMID- 15539202 TI - Signal transduction in insulin action. PMID- 15539203 TI - Dietary trans fatty acids modulate erythrocyte membrane fatty acyl composition and insulin binding in monkeys. AB - The substitution of trans- for half of the cis-monounsaturated fatty acids in the diet of Macaca fasicularis monkeys resulted in alterations in erythrocyte fatty acid composition and insulin receptor properties but not in membrane fluidity. Both cis and trans diets contained 10% fat and similar fatty acid compositions, except that approximately 50% of the cis-octadecenoate (c-18:1) in the cis diet was replaced with trans-octadecenoate isomers (t-18:1) in the trans diet. Compared with the cis diet, the trans diet resulted in the incorporation of approximately 11% t-18:1, an approximately 50% decrease in c-18:1, an approximately 16% decrease in total saturated fatty acids, and an approximately 20% increase in 18:2(n-6) in erythrocyte membrane lipids. The increase in 18:2(n 6) may reflect on homeostatic mechanisms designed to maintain overall membrane fluidity, as no diet-related changes in fluidity were observed with diphenylhexatriene steady state fluorescence polarization. Values observed for insulin binding and insulin receptor number were higher and binding affinity was lower in monkeys fed the cis diet. In the absence of an effect on overall membrane fluidity, altered receptor activity suggests that insulin receptor activity is dynamic, requiring specific fluid membrane subdomains or highly specific fatty acid-protein interactions. PMID- 15539204 TI - Nutritional characteristics of a neoglycoprotein, casein modified covalently by glucose. AB - Glucose was combined covalently with the epsilon-amino groups of lysyl residues of bovine casein in the presence of sodium cyanoborohydride as a reducing reagent by reductive alkylation, forming stable secondary amine linkages. Solubility characteristics and nutritional values of the neoglycoprotein were examined. The degree of modification (%) of the glucosylated casein was 82.5. Solubility of the modified casein was increased by the attachment of glucose. The modification did not disturb the digestion of casein by pepsin or trypsin. Rat feeding experiments using 10% protein diets demonstrated that the protein efficiency ratio (PER) of the modified casein was 0.35 +/- 0.33 compared with 2.99 +/- 0.29 for the unmodified casein. When the modified casein was supplemented with L-lysine to equal the level of total lysine of unmodified casein, the PER value was increased to 2.21 +/- 0.29. Nitrogen balance experiments showed that the modified casein was digested completely. On the other hand, biological value and net protein utilization of the modified protein were shown to be considerably lower than those of the unmodified casein. PMID- 15539205 TI - Effect of prostaglandin inhibition on caffeine-induced hypercalciuria in healthy women. AB - Caffeine ingestion increases urinary calcium excretion. The mechanism is not known, but prostaglandin synthesis has been implicated. We hypothesized that administration of a prostaglandin inhibitor such as acetylsalicylic acid (aspirin) along with caffeine would prevent caffeine-induced hypercalciuria. We measured 3-hour excretion in fasting subjects who each randomly ingested four treatments on nonconcurrent mornings: no drug, caffeine (5 mg/kg body weight), acetylsalicylic acid (650 mg), or caffeine plus acetylsalicylic acid. In experiment 1, nine healthy premenopausal female subjects were studied; each treatment was taken with 200 ml of orange juice. Water was provided hourly to encourage urine flow. Urinary calcium excretion rose with caffeine treatment; mean 3-hour calcium (mmol/mmol creatinine) was 0.49 +/- 0.07 compared with 0.23 +/- 0.04 during the no-drug treatment. Acetylsalicylic acid caused a significant reduction in urinary calcium to 0.13 +/- 0.08; when it was combined with caffeine, caffeine-induced calcium excretion fell significantly to 0.35 +/- 0.08. Sodium excretion tended to reflect calcium excretion. Urinary prostaglandin E(2) fell significantly with acetylsalicylic acid, with and without caffeine. There were no significant changes in creatinine, water, or potassium excretion. Experiment 2 was similar, except that water was substituted for orange juice to test the possibility that acetylsalicylic acid affected elevated but not basal calcium excretion. Similar and even more pronounced results were obtained, with caffeine causing a threefold increase in urinary calcium, acetylsalicylic acid causing a decrease by half, and the combined drug treatment being greater than no drug but less than caffeine alone. Urinary phosphorus rose significantly with caffeine alone. Prostaglandin synthesis may not be directly involved in caffeine induced hypercalciuria, as the magnitude of the caffeine-induced increase was similar when treatments given the acetylsalicylic acid were compared with those without a prostaglandin synthesis inhibitor. PMID- 15539206 TI - Effect of dietary lipid on collagen- and adenosine diphosphate-induced platelet aggregation and thromboxane A2 synthesis in the rat. AB - Dietary lipids containing different proportions of long-chain polyunsaturated fatty acids can affect platelet thromboxane A(2) formation and aggregation. In the present work, the effects of dietary lipid, from animal and vegetable sources, on collagen- and adenosine diphosphate (ADP)-induced thromboxane A(2) (measured as thromboxane B(2)) production and aggregation in washed rat platelets were studied. In addition, plasma thromboxane B(2) levels in rats fed different dietary lipids were measured. Animals were fed 10% fat by weight as lard (LRD), corn oil, soy bean oil, canola oil (CAN), or cod liver oil (CLO) for a period of 7 weeks. Circulating thromboxane B(2) levels detected in platelet-poor plasma of the CLO-fed animals were significantly lower than those of rats fed all other dietary lipids. The platelets of CLO-fed animals synthesized significantly less thromboxane A(2) compared with those from other dietary groups following ex vivo stimulation of platelets with agonists such as collagen and ADP, with the exception of platelets from the LRD-fed animals. Ex vivo stimulation of platelets obtained from this group with collagen resulted in the synthesis of significantly greater levels of thromboxane A(2) compared with all other groups. However, aggregation responses to collagen and ADP were not significantly affected by dietary treatment, although relatively the lowest responses to these agonists were apparent in the CLO-fed and CAN-fed groups, respectively. PMID- 15539207 TI - Effect of lectins on the cobalamin-protein binding reactions: implications for the tissue uptake of cobalamin. AB - Plant lectins have been thought to impair nutrient absorption, both by specific and nonspecific interference in the absorptive process. The main objective of this investigation was to study the effect of lectins on the various binding reactions involving cobalamin (cbl)-protein complexes and their receptors, and to identify the rate-limiting step important in maintaining tissue levels of cobalamin. Among the lectins tested in vivo, only concanavalin A (ConA) was able to inhibit the transport of cobalamin to the tissues and caused a 70% to 75% inhibition of [(57)Co] cobalamin transported to the liver and kidney. The inhibition of transport to the tissues was independent of route of administration of cobalamin, whether intragastric or systemic, and was not due to decreased gastrointestinal uptake. When tested in vitro, concanavalin A inhibited the binding of transcobalamin II-cbl to its receptor, but not the binding of cobalamin to intrinsic factor or intrinsic factor-cobalamin complex to the ileal receptor. These results suggest that late events during transcellular transport of cobalamin through the enterocytes is the rate-limiting step determining tissue levels of cobalamin and that ConA inhibits these latter events. PMID- 15539208 TI - Plasma tryptophan and tyrosine concentrations: determination using high performance liquid chromatography and fluorometric detection. PMID- 15539209 TI - Methionine metabolism in mammals. PMID- 15539210 TI - Characterization of transferrin receptors on plasma membranes of lactating rat mammary tissue. AB - Little is known about the transport of iron into the mammary secretory cell and the process of milk iron secretion. The concentration of iron in milk is remarkably unaffected by maternal iron status, suggesting that the uptake of iron into the mammary gland is regulated. It is known that iron enters other cells via transferrin receptor-mediated endocytosis. This study was designed to isolate and characterize the mammary gland transferrin receptor in lactating rat mammary tissue using immunochemical techniques. The existence of functional mammary gland transferrin receptors in lactating rodents was demonstrated using radiolabel binding techniques. Isolation of mammary transferrin receptors by affinity chromatography was confirmed using immunoelectrophoresis and slot blot analysis. The intact transferrin receptor was found to have a molecular weight of 176 kd as determined by Western blotting followed by scanning densitometry. Reduction of the receptor with beta-mercaptoethanol gave a molecular weight of 98 kd. An additional immunoreactive band of 135 kd was observed. The presence of transferrin receptors in normal lactating rat mammary tissue is likely to explain iron transport into mammary tissue for both cellular metabolism and milk iron secretion. PMID- 15539211 TI - Thyroxine effects on parameters of glucose turnover in BHE rats fed menhaden oil. AB - The effect of hyperthyroidism on glucose turnover in BHE rats fed menhaden oil was studied. Thyroxine-treated rats had a greater glucose mass, a greater absolute glucose synthesis rate, less hepatic and muscle glycogen levels, and greater hepatic and peripheral fat cell lipogenic rates than nontreated rats. No differences in body weight gain were observed, nor were there differences in blood glucose levels, glucose space, or fractional reversible or irreversible glucose use. These observations suggest that thyroxine and menhaden oil were additive in their effects on glucose metabolism in BHE rats, which are genetically programmed to develop non-insulin-dependent diabetes mellitus. PMID- 15539212 TI - Development of the stable isotope tracer approach for studies of copper turnover in the rat and mouse. AB - The stable isotope tracer approach was explored for long-term investigations of copper turnover in the adult rat and mouse, with inductively coupled plasma mass spectrometry for isotope measurements. The isotopic measurement method permitted precision and accuracy of <1.0%, with an overall sample blank of <0.05 microg copper. Rats were fed a copper-deficient diet and deionized water with (+Cu) or without (-Cu) copper (20 microg/ml). Both groups underwent a single-day replacement of drinking water with 20 microg/ml of (65)Cu. Compared with the baseline isotope ratio ((65)Cu/(63)Cu) of 0.462 +/- 0.002, blood plasma ratios for the +Cu group on days 2, 7, and 14 postdosing were 0.702 +/- 0.021, 0.557 +/- 0.004, and 0.474 +/- 0.001, respectively. The corresponding data for liver were 1.652 +/- 0.018, 0.560 +/- 0.005, and 0.482 +/- 0.001, respectively. For the -Cu group, respective plasma ratios were 1.580 +/- 0.04. 0.917 +/- 0.02, and 0.664 +/ 0.01 for days 2, 7, and 14 postdosing, and the ratios for liver were 0.987 +/- 0.02, 0.876 +/- 0.04, and 0.739 +/- 0.03. Mice previously made copper deficient to varying degrees were given a single-day replacement with the label. When the 24-hour postdosing isotope ratios in the livers of these mice were correlated with the activity of plasma ceruloplasmin, a negative correlation (r = -0.85) was observed. Isotope enrichment in both rats and mice was greater in the copper deficient animals compared with the controls. PMID- 15539213 TI - Folate binding in intestinal brush border membranes: evidence for the presence of two binding activities. AB - Binding of [(3)H]folic acid by isolated rat jejunal brush border membranes (BBMs) was analyzed by chromatography on small Biogel P-30 columns. Folic acid binding to BBMs exhibited a prominent pH effect with a sharp maximum at pH 5.5 to 6.0. After acid treatment to strip the BBMs of bound folate, the membranes demonstrated a wider pH optimum (5.5 to 7.5) of folate binding and a higher binding capacity. Scatchard analysis of binding experiments performed at pH 6.0 revealed the existence of two components: one with a high affinity (kd = 12 to 25 nM) and low capacity (V(max) for non-acidified BBMs = 0.259 to 0.264 pmol/mg protein, V(max) for acidified BBMs = 0.41 to 0.71 pmol/mg protein) and the other with a low affinity (kd = 1.1 to 5.1 microM and high capacity (V(max) for non acidified BBMs = 0.93 to 1.93 pmol/mg protein, V(max) for acidified BBMs = 4.05 to 7.69 pmol/mg protein). Phosphatidylinositol-specific phospholipase C preferentially detached the high affinity component from jejunal BBMs. Phosphatidylinositol-specific phospholipase C-released folate binding protein was precipitated by antibodies to the high-affinity folate-binding protein from rat kidney. These data suggest the existence of two different folate-binding proteins in isolated rat jejunal BBMs. The high-affinity folate-binding protein shares epitopes with the folate-binding protein in the kidney. PMID- 15539214 TI - Monoclonal antibody studies of the antigenic determinants of human plasma retinol binding protein. AB - A battery of monoclonal antibodies (MoAbs) against human retinol-binding protein (RBP) was produced to obtain useful probes for the study of the antigenic determinants of RBP. The 12 antibodies all reacted with human RBP by immunoblotting. Based on antibody cross-competition radioimmunoassays, four distinct and different groups of antibodies were identified: group I, 1A4 and 2F4; group II, 1G10, 5C5, 6F4, and 7G3; group III, 5H6, 6C7, 10G5, and 14E3; and group IV, 5H9 and 13A1. Information about the epitopes of RBP recognized by these MoAbs was obtained by testing the reactivity of each antibody with human, rabbit, and rat RBPs by immunoblotting. Group I and group IV antibodies reacted to a similar extent with human, rabbit, and rat RBPs. Group II antibodies reacted strongly with human and rabbit RBPs, but reacted very weakly with rat RBP. Group III antibodies reacted strongly with human RBP, but did not react with rabbit or rat RBP. Thus, the epitopes for group I and group IV antibodies appear to be regions of the RBP molecule that are conserved across the three species, whereas group III antibodies recognized only human RBP. In a preliminary study, the reactivity of each antibody with purified cyanogen bromide fragments of RBP was tested by slot immunoblotting. None of the MoAbs reacted with any of the cyanogen bromide fragments. This study shows that MoAbs specific for at least four different regions of the RBP molecule can be produced; hence, RBP contains at least four major antigenic domains. PMID- 15539216 TI - Recent progress in vitamin A research: nuclear retinoic acid receptors and their interaction with gene elements. PMID- 15539215 TI - A method for the determination of the circulating concentration of vitamin D. PMID- 15539217 TI - Specific binding and uptake of apolipoprotein E-free high density lipoproteins by cultured liver parenchymal cells of copper-deficient rats. AB - The influence of copper deficiency on the binding and uptake of apolipoprotein E free high density lipoprotein (apo E-free HDL) in cultured rat hepatic parenchymal cells was examined in this study. Male weanling Sprague-Dawley rats were randomly divided into two treatments, a Cu-adequate (7.33 mg Cu/kg diet) or a Cu-deficient (1.04 mg Cu/kg diet) group. After 7 weeks, plasma apo E-free HDL were isolated by a combination of ultracentrifugation, gel filtration, and heparin-Sepharose affinity chromatography. Parenchymal cells were isolated from collagenase perfused liver of Cu-deficient and adequate rats and cultured for 16 hours at 37 degrees C prior to incubation with iodinated apo E-free HDL from the same treatment group. Cells were incubated with 5 microg/ml(125) I-apo E-free HDL for 2, 6, or 12 hours in the presence or absence of 200 microg/ml (40-fold) excess unlabeled apo E-free HDL. Increases in specific binding at 4 degrees C and specific cell-associated uptake at 37 degrees C as a function of time were observed with cells and HDL from Cu-deficient rats. Cells were also incubated for 6 hours with 8 concentrations of (125)I-apo E-free HDL in the presence or absence of excess unlabeled HDL. Although no significant increase in specific binding was detected at 4 degrees C as a function of ligand concentration, the response tended to be higher at 5 to 15 microg HDL/ml for the Cu-deficient treatment. However, at 37 degrees C the specific cell-associated uptake was increased markedly with cells and HDL from Cu-deficient rats. The observed increases in HDL binding and uptake indicate that these processes may be enhanced in Cu-deficient rats. These data are also consistent with recent in vivo results which indicate that plasma clearance and tissue uptake of HDL are increased in Cu-deficient rats. PMID- 15539218 TI - Changes in fatty acid composition of rat liver and serum induced by distal small bowel resection. AB - The serum lipid composition and the fatty-acid profiles of the major lipid fraction (triglycerides, esterified cholesterol, and phospholipid) of liver and serum were examined 6 weeks after both 50% and 75% distal small bowel resection (DSBR). Total serum lipid content did not modify after DSBR. Esterified cholesterol and phospholipid levels of the serum did not significantly change after the operation. However, a significant increase in both free cholesterol and triglyceride levels was observed after DSBR. Different fatty acid changes in the liver and serum lipid fractions were found after DSBR, with the greatest differences in the hepatic esterified cholesterol fraction. These results suggest that DSBR affects both the lipid composition and the fatty acid composition of major lipid fraction of liver and serum. PMID- 15539219 TI - Dietary lipid effects on microsome fatty acid composition of liver and brain, on liver glucose-6-phosphatase, and on brain 5'-nucleotidase activity in the rat. AB - The effects of incorporation of dietary oils with different n6/n3 ratio and polyunsaturated fatty acids content into rat liver and brain microsomes has been studied. The investigation of membrane fatty acid composition of liver microsomes and that of brain microsomes gave different results. In particular, liver microsomes of rats fed fish oil showed a relatively higher content of 20:5n3 and 22:6n3, and a lower content of 20:4n6. Under these conditions, a reduced glucose 6-phosphatase activity was measured. Brain microsomal fatty acid composition was only slightly affected by dietary lipid intake. The 5'-nucleotidase activity of those particles was similar, although statistically different values were found in fish-oil-fed rats and in olive-oil-fed rats. The effects of membrane fatty acid composition on membrane-bound enzyme activity are discussed. PMID- 15539220 TI - Tissue iron content in riboflavin and pyridoxine deficient rats. AB - The effect of riboflavin and (or) pyridoxine deficiency and repletion on tissue iron content was studied in rats. The iron content in liver, spleen, and kidney and plasma iron concentration of riboflavin deficient (RD) rats was lower, but hematocrit was not. In pyridoxine deficient (PD) rats versus control rats, the iron content in liver was significantly higher but not in spleen and kidney. In PD rats hematocrit was lower but plasma iron concentration was not. Although combined riboflavin and pyridoxine deficient (CD) rats had lower iron content in liver and spleen compared with control rats, these values were intermediate between those of RD rats and PD rats. After RD and PD rats were repleted, the iron content in liver, spleen, and kidney returned to that of control rats, and the hematological indices were improved significantly. These results suggest that riboflavin and pyridoxine deficiency may impair the absorption and utilization of iron and may result in altered tissue iron content. PMID- 15539221 TI - Methods for the determination of the circulating concentration of 25 hydroxyvitamin D. PMID- 15539222 TI - A method for the determination of the circulating concentration of 1,25 dihydroxyvitamin D. PMID- 15539223 TI - Choline deficiency. PMID- 15539224 TI - Effects of two prototypic polychlorinated biphenyls (PCBs) on lipid composition of rat liver and serum. AB - Mature male Sprague-Dawley rats received a single IP injection of either 2,2',4,4',5,5'-hexachlorobiphenyl (HCB), 3,3',4,4'-tetrachlorobiphenyl (TCB) (300 microm/kg) in corn oil (10 ml/kg) or the corn oil vehicle alone, and were killed four days later after having been fasted overnight. The vehicle control group consisted of rats which were allowed free access to feed as well as pair-fed animals. Lipid analyses were conducted on liver, hepatic microsomes and serum. TCB- (but no HCB-) treatment resulted in a statistically significant increase in total liver lipids and triglycerides. Liver phospholipids remained unchanged. Both PCBs increased the cholesterol and phospholipids content of the liver microsomal fraction. Serum lipids measured were not statistically different from control values. While HCB had little effect on the fatty acid composition of liver lipids, TCB caused an increase in C 18:1 (n-9) and a decrease in C 20:4 (n 6). Both PCBs increased C 18:0 in the hepatic microsomal fraction, but TCB also decreased C 16:0. Neither PCB altered the fatty acid composition of serum total lipids. These data are consistent with the concept that specific alterations in lipid metabolism are dependent on the structure of the PCB. PMID- 15539225 TI - Effect of Mg nutriture on the dynamics of administered 25Mg exchange in vivo in the rat. AB - The effect of Mg nutriture on Mg exchange and interorgan distribution was studied in adult rats ten days after a single I.P. dose of (25)Mg ( approximately 5 mg). First the effects of level of Mg intake (0.25, 0.05, or 0.01% Mg) on standard measures of Mg nutriture were studied for 62d to fully document the Mg status of the adult rats. The Mg-deficient diet led to a reduction in plasma, erythrocyte and urine Mg concentration but the only tissues affected were kidney and bone; no outward signs of deficiency were observed. At this point, the 4 remaining rats from each diet group received a single dose of (25)Mg and were killed 10d later. Unlike measures of total Mg content, Mg restriction was observed to significantly alter the distribution of isotope within the soft tissue compartment. The proportion of retained isotope accumulated by soft tissues other than skeletal muscle increased. Because this was not true for skeletal muscle, exogenous (25)Mg label was diverted to more metabolically active tissues during Mg restriction. The apparent Mg exchangeable pool (MgEP) size, determined by in vivo stable isotope dilution, reflected this difference in skeletal muscle (25)Mg accumulation; MgEP size was 39% lower in Mg restricted (0.01% Mg) compared to control (0.05% Mg) rats. The pool of exchangeable Mg in bone was also reduced by Mg restriction but, unlike the soft tissue compartment, the reduction in bone exchangeable Mg was quantitatively similar to the reduction in total Mg content. PMID- 15539226 TI - Dietary copper, simple sugars, and metabolic changes in pigs. AB - Inadequate dietary copper is known to result in undesirable metabolic changes in rats and humans. Abnormal cardiac function, leading to sudden death, is a common finding when copper deficient rats are fed a 62% fructose diet. To further study the apparent mineral-carbohydrate relationship to cardiac physiology, 3 male and 3 female swine were randomly assigned to four groups (6 pigs per group) which were fed low copper (1.5 ppm) or copper supplemented (40 ppm) diets with 20% of calories from either fructose or glucose for 10 weeks. In agreement with results from other animal studies, copper deficient swine exhibited decreased plasma ceruloplasmin, erythrocyte superoxide dismutase and plasma lysyl oxidase activities and lowered serum copper. The copper deficient fructose group had the lowest aortic lysyl oxidase activity and hematocrit when compared to the other groups. The relative heart weight in the copper deficient fructose group was 93% greater than the other three dietary groups. The livers of copper deficient fructose fed pigs were also significantly larger. Two enzymes related to cardiac and hepatic function, aspartate and alanine aminotransferase were also measured. Copper deficiency significantly lowered alanine aminotransferase but there was no dietary effect on aspartate amino-transferase. The results of this project indicate that the pig is a sensitive model for the study of cardiovascular abnormalities which occur when fructose is consumed with a low copper diet. PMID- 15539227 TI - Lysine flux in dry and lactating dairy goats. AB - The objective of this experiment was to test the hypothesis that the physiological state of lactation is accompanied by both an increase in total plasma lysine flux (rate of loss and replacement of lysine) and a net reduction in flux through the plasma lysine pool after accounting for lysine secreted in the milk. Eight lactating French Alpine does were primed and infused for three hours with solutions of alpha15N L-lysine HCl in 0.9% saline through indwelling jugular vein catheters. Enrichment of circulating plasma lysine by continuous intravenous infusion of alpha15N L-lysine was used to estimate whole body lysine flux. This procedure was repeated one month after cessation of milking. Total plasma lysine flux was similar for dry and lactating does (116.6 and 123.0 mmol/d, SEM 16.6 mmol), but 54.2 mmol/d lysine was secreted as milk protein during lactation. Direct measurement of lysine absorption from the lower tract and independent measurement of lysine degradation are needed to provide a more complete portrait of caprine lysine kinetics. PMID- 15539229 TI - Quantitative analysis of fatty acids in phospholipids, diacylglycerols, free fatty acids, and other lipids. PMID- 15539228 TI - Effect of cis-diamminedichloroplatinum (II) on metallothionein induction and trace element metabolism in rats fed different amounts of dietary zinc. AB - Recent studies have suggested that the induction of metallothionein synthesis in kidneys of mice by the acute administration of bismuth and other trace elements might protect against cis-diamminedichloroplatinum (II) nephrotoxicity. The present study was designed to determine the effects of dietary zinc and cis diamminedichloroplatinum (II) on the induction of liver and kidney metallothionein and its subsequent effect on nephrotoxicity and trace element metabolism in rats. Male rats were fed diets containing 5, 20, 80, or 320 mg zinc/kg diet for 3 weeks. Each dietary group was subdivided into 3 groups. In one group, each rat received an i.p. injection of 7.5 mg cis-diamminedichloroplatinum (II)/kg b.w. All other rats received saline. During the next three days a second group of rats was pair-fed to the cis-diamminedichloroplatinum (II) injected group. A third group received no treatment and was allowed to eat ad libitum. Results showed that when dietary zinc was increased from 5 mg/kg diet to higher amounts, kidney metallothionein concentration increased twofold. cis diamminedichloroplatinum (II) treatment increased kidney metallothionein even further, but elevated metallothionein gave no protection from the toxic effects of the drug. Serum copper concentration and ceruloplasmin activity were significantly lower with higher concentrations of dietary zinc, which indicated that these rats were mildly copper-deficient. There was a small but significant depression of superoxide dismutase activity and a highly significant increase in thiobarbituric acid reactive substances in kidneys of rats treated with cis diamminedichloroplatinum (II) compared to either pair-fed or ad libitum controls. This supports the hypothesis that part of the mechanism for cis diamminedichloroplatinum (II)-induced toxicity might be caused by free-radical generation. However, the data do not support the hypothesis that metallothionein induction protects the kidney from cis-diamminedichloroplatinum (II) toxicity. PMID- 15539230 TI - Mitochondrial enzymes responsible for oxidizing medium-chain fatty acids in developing rat skeletal muscle, heart, and liver. AB - Prior to weaning, medium-chain fatty acids constitute an important energy source in the developing rat. Fatty acid oxidation rates increase with age in most developing tissues, but the pattern of this increase may vary according to the role of the particular organ. In skeletal muscle, heart, and liver of developing rats, we measured mitochondrial activities of long- and short-chain enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and long- and short-chain acyl-CoA thiolase. In skeletal muscle, the pattern of development in fatty acid oxidation enzymes favored utilization of long-chain rather than medium-chain fatty acids. In liver, enzyme activities for medium-chain fatty acids were highest prior to weaning. Heart occupied a position intermediate between skeletal muscle and liver. PMID- 15539231 TI - Development of copper deficiency in neonatal mice. AB - Dietary copper (Cu) deficiency was produced in Swiss albino mice to determine the temporal relationship between depletion of Cu and changes in the cardiovascular and nervous system. Dams were placed on a Cu-deficient diet 4 days after parturition. Half the dams were provided with deionized water and their offspring are referred to as Cu-deficient (-Cu). Half the dams were given cupric sulfate in their drinking water (20 microg Cu/mL) and their offspring are referred to as Cu adequate (+Cu). At 6 weeks of age a sample of the -Cu mice were repleted with CuSO(4). Mice were sampled 1 day after birth and at weekly intervals for 7 weeks. Both +Cu and -Cu mice grew at the same rate: birth weight increased 16-fold at 6 weeks of age. Liver Cu more than doubled between 1 and 7 days of age. At 2 weeks of age -Cu mice were anemic (lower hematocrit and hemoglobin) and had lower liver Cu and plasma ceruloplasmin activity compared to +Cu mice. Liver Fe was not elevated in -Cu mice until 2 weeks after anemia developed. At weaning first signs of altered catecholamine metabolism included elevation of dopamine in both heart and spleen. Norepinephrine concentrations and content, in contrast, were not both lowered in -Cu mice until 5 weeks of age. Heart weight was first elevated in -Cu mice at 6 weeks of age and relative weight (mg/g body wt) at 4 weeks of age. Liver Cu concentration was lower in 1-week repleted mice than in +Cu mice. Anemia preceded the development of cardiac hypertrophy and altered catecholamine levels in -Cu mice. PMID- 15539232 TI - Effects of bile salt supplementation on biliary secretion in estrogen-treated rats. AB - The aim of the present study was to determine whether bile acid feeding to rats can reverse ethinyl estradiol-induced cholestasis. Animals received ethinyl estradiol (2 mg/kg/day) for 6 days or were coinfused with estrogen plus various bile acids (60 mg/kg/day). Cholestasis could be significantly prevented by tauroursodeoxycholic acid, was partly corrected by ursodeoxycholic acid, and was unchanged by chenodeoxycholic acid. Total bile salt secretion was increased in every group. The secretion of the major primary bile acids (cholic acid and beta muricholic acid) was restored to a large extent in rats supplemented with tauroursodeoxycholate but not in chenodeoxycholate-fed rats. In the former group, the canalicular transport of taurocholate and the bile salt pool size were identical with those of control rats. The hydrophilic-hydrophobic balance of the administered bile salt species appears to be an essential factor in the restoration of bile secretion, the more hydrophilic bile salt having the more hepatoprotective effect. PMID- 15539233 TI - Effect of selenium deficiency on tissue taurine concentration and urinary taurine excretion in the rat. AB - The purpose of this study was to determine the effect of selenium deficiency on tissue taurine levels and urinary taurine excretion. Weanling male Sprague-Dawley rats were fed selenium-deficient or selenium-adequate diets for 20 weeks. As selenium deficiency developed, urinary taurine excretion increased in selenium deficient rats compared to controls. At 12 weeks, the selenium-deficient rats excreted 1.7-fold more taurine than control rats. At the same time plasma glutathione peroxidase was 1.2% of control and plasma glutathione was 226% of control. At 20 weeks, renal taurine was decreased but renal glutathione was increased in selenium-deficient rats compared to controls. Feeding the experimental diet for 6 weeks without methionine supplementation caused a fall in urinary taurine excretion. However, there was no difference between selenium deficient and control rats. These results indicate that selenium deficiency affects renal handling of taurine in the rat when dietary sulfur amino acids are not restricted. PMID- 15539234 TI - Choline deficiency activates phospholipases A2 and C in rat liver without affecting the activity of protein kinase C. AB - There is evidence to suggest that liver tumor promoters exert their effect through the interference of signal transduction in hepatic cells. Both phospholipase A(2) and phospholipase C play important roles in the generation of second messengers and in the activation of Ca(2+), phospholipid-dependent protein kinase C. Using male Sprague-Dawley rats, we investigated whether liver tumor promoting regimens of a choline-deficient diet and phenobarbital alter the activities of phospholipase A(2) and phospholipase C in the liver, and extended the study to determine the effect of a choline-deficient diet on protein kinase C activities. Feeding a choline-deficient diet for 1 week increased the activities of both phospholipase A(2) (50%) and phospholipase C (22%), and the activities of both enzymes were more than doubled after 4 weeks. Feeding a phenobarbital diet resulted in a slight decrease in phospholipase A(2) activities at 4 weeks but no significant changes in PLC activities. The protein kinase C activities and its distribution between soluble and particulate fractions remained unchanged after 1, 2, and 4 weeks feeding of a choline-deficient diet. Thus, activation of both phospholipase A(2) and C is distinct for a choline-deficient diet, not shared by phenobarbital diet. Increased activities of these enzymes were not associated with the activation of protein kinase C under the present experimental condition. PMID- 15539235 TI - Adenosine diphosphate and adenosine monophosphate: enzymatic spectrophotometric determination. PMID- 15539236 TI - Biochemical changes in copper deficiency. PMID- 15539237 TI - Metabolic effects of medium chain triglyceride-enriched total parenteral nutrition in rats bearing Yoshida sarcoma. AB - The ability of medium chain triglyceride-enriched total parenteral nutrition to support host tissue in a model of cancer cachexia was assessed by measuring tumor growth, body weight, nitrogen balance, energy expenditure, leucine kinetics, fractional protein synthetic rate of tumor, liver, and abdominis rectus muscle, and plasma levels of glucose and albumin. Male Sprague-Dawley rats (85-90 gm) received 10(7) cells of viable Yoshida sarcoma subcutaneously on day 0. Control rats received injections of sterile saline. On day 10 rats underwent central venous cannulation and were randomized to one of three isocaloric diets. One group received amino acids and dextrose, while the other two groups were infused with amino acids, dextrose, and fat as either long chain triglyceride or a physical mixture of medium chain triglyceride: long chain triglyceride (3:1). On day 14 L-1-(14)C-leucine was added to the diet to study protein kinetics, and energy metabolism was measured by indirect calorimetry. Both tumor-bearing and nontumor-bearing rats demonstrated improved nitrogen balance when given medium chain triglyceride-enriched total parenteral nutrition. Tumor-bearing rats had reduced resting energy expenditure vs. nontumor-bearing, while rats receiving total parenteral nutrition without fat had significantly greater respiratory quotients. Tumor-bearing rats had lower total body weight vs. nontumor-bearing on day 10, but body weight of tumor-bearing and nontumor-bearing did not differ on day 14. Whole body protein breakdown decreased and leucine balance increased in tumor-bearing rats as compared to nontumor-bearing. Total liver mass was greater in tumor-bearing rats, but liver protein fractional protein synthetic rate decreased in tumor-bearing rats vs. nontumor-bearing. Tumor growth rate and fractional protein synthetic rate were not altered by the parenteral diet. The data confirm an altered metabolism in the tumor-bearing host, and suggest that medium chain triglyceride can better support host tissue. PMID- 15539238 TI - Effects of dietary n-3 fatty acids on the composition of cholesteryl esters and triglycerides in plasma and liver perfusate of the rat. AB - The effects of polyunsaturated fatty acids of the omega-3 family (PUFA n-3), (addition of fish oil), on the molecular composition of cholesteryl esters and triglycerides in plasma and liver perfusate of rats were studied. Rats fed a diet rich in saturated fatty acids (addition of lard) served as controls. Supplemention with PUFA n-3 not only decreases the plasma concentrations of free cholesterol, cholesteryl esters, and triglycerides, it also significantly alters the plasma composition of cholesteryl esters and triglycerides. Analyses of liver perfusate indicate a decrease in triglycerides secretion by in vitro perfused liver and reciprocal changes in relative contents of cholesteryl esters fractions with C(16) and C(20) acyl chains. This finding may be a result of chain shortening of long-chain fatty acids probably in peroxisomal beta-oxidative system. Alterations in plasma cholesteryl esters and triglycerides composition of the fish oil group could be affected further by additional factors such as increased plasma cholesterol esterification activity and presence of triglyceride species of intestinal origin. PMID- 15539239 TI - Consequences of selenite supplementation on the growth and metabolism of cultures of canine mammary cells. AB - Previous studies with cultures of canine mammary cells revealed differences in the degree of growth inhibition caused by selenite supplementation, with canine mammary tumor cell line 13 > 11 >> non-neoplastic canine mammary cells. The present studies show this variation in growth retardation cannot be explained by selenium retention. Intracellular glutathione related inversely to the degree of growth inhibition resulting from the addition of selenite. Dimethyl selenide formation by S-9 preparations corresponded to the sensitivity of the culture to supplemental selenite. DL-buthionine-SR-sulfoximine, a specific inhibitor of glutathione biosynthesis, accentuated the growth inhibition and prevented the increase in intracellular glutathione caused by supplemental selenite. Treatment of canine mammary tumor cell line 13 cultures with DL-buthionine-SR-sulfoximine resulted in a persistent depletion of intracellular glutathione without affecting growth. Glutathione reductase activity, before and following selenite, was inversely related to the degree of growth inhibition, with canine mammary tumor cell line 13 > 11 > non-neoplastic canine mammary tumor cell line. Selenite addition increased the activity of gamma-glutamylcysteine synthetase in canine mammary tumor cell line 11 and non-neoplastic canine mammary cells, but not in canine mammary tumor cell line 13 cells. The present data suggest the differences in the growth inhibition caused by selenite among these mammary cells is related to glutathione regulation and ultimately to selenium detoxification. PMID- 15539240 TI - Effect of low-calorie diets on plasma retinol-binding protein concentrations in overweight women. AB - The concentrations of total protein, albumin and retinol-binding protein, a major transport protein for vitamin A, are significantly decreased by protein-calorie malnutrition. Weight-loss diets, sometimes involving severe energy deficits over prolonged periods of time, are common in the United States. The effect, if any, of prolonged low calorie weight-loss diets with normal intakes of protein on albumin, total protein and retinol-binding protein concentrations (and potentially on vitamin A metabolism) had not been extensively studied. We measured total protein, albumin, apo + holo retinol-binding protein and holo-free and holo-transthyretin-bound retinol-binding protein concentrations during the course of a nutritionally adequate weight-loss diet (50% calorie restriction). We found that this type of dieting did not affect total protein, albumin or apo + holo, holo-free or holo-transthretin-bound retinol-binding protein concentrations significantly. This suggests that protein intake is more critical than caloric intake for retinol-binding protein status. PMID- 15539241 TI - Silicon facilitation of copper utilization in the rat. AB - An eight-week, 2 x 4 factorial rat experiment using two levels of dietary copper and four levels of dietary silicon was conducted to further delineate a previously observed silicon-copper interaction in which silicon appears to mimic copper in its effect on the composition of the aorta. Dietary copper concentrations were 1.4 (deficient) and 5.4 (adequate) mg/kg diet, and silicon concentrations were 5, 135, 270, and 540 mg/kg diet. Compared with the lowest level of silicon and copper, weight gains were 15.5% higher for rats fed 540 mg silicon/kg diet and 14.3% higher for those fed 5.4 mg copper/kg diet. The growth promoting effects of silicon and copper were additive. Evidence that silicon elevated the copper status of copper-deficient rats includes an increase in packed-cell volume by 540 mg silicon/kg diet in the otherwise packed-cell volume depressed, copper-deficient rats, accompanied by a trend toward higher hemoglobin values and lower relative heart weights. In the copper-adequate rats, evidence that 540 mg silicon/kg diet elevated their copper status includes a two-fold increase in the blood-plasma copper concentration, a three-fold increase in ceruloplasmin activity, and an increase in cardiac, renal, and hepatic copper concentrations. In addition, 540 mg silicon/kg diet resulted in higher aortic dry mass and aortic elastin content in both copper-deficient and copper-adequate rats. While dietary silicon concentrations of 135, 270, and 540 mg/kg diet were all effective in increasing aortic elastin in the copper-adequate rats, only 540 mg silicon/kg diet increased aortic elastin in the copper-deficient rats. These data indicate that some of the metabolic effects attributed to silicon may be manifested through a silicon-facilitated increase in copper utilization. PMID- 15539242 TI - Analysis of lipids by high performance liquid chromatography: part I. PMID- 15539243 TI - Indications for total parenteral nutrition in the hospitalized patient: a prospective review of evolving practice. AB - The indications for initiating total parenteral nutrition (TPN) were prospectively evaluated in 100 consecutive patients at a tertiary referral hospital with a long-standing Nutritional Support Service to illustrate the reasons why the parenteral route was chosen at this unique institution in terms of patient population. Sixty male and 40 female patients, average age 59 +/- 17 years (range 22-86 years), were classified a priori as to the underlying reasons for initiation of TPN. The study was conducted by a Nutrition Support Service at this hospital without pediatric, trauma, or burn services specializing in the care of patients with diabetes mellitus. Of the 100 patients, 63% were from the surgical service; 24% had diabetes mellitus. Their mean weight (118 +/- 29% of ideal), body mass index (25 +/- 6 kg/m(2)), and serum albumin (2.8 +/- 0.7 g/dL) indicated a reasonable body composition with a moderate systemic inflammatory response. Six patients received preoperative TPN for an average of 5 +/- 3 days with a variety of diagnoses including malignancy, Crohn's disease, bowel obstruction, and gastrointestinal bleeding. The underlying reasons for initiating nutritional support were related to three factors that largely determine the need for involuntary feeding: preexisting protein calorie malnutrition, actual or anticipated semistarvation for a prolonged period, and the presence of a systemic inflammatory response. The choice of TPN was based on anticipated or proven intolerance to full enteral feeding. The duration of time before initiation of TPN postoperatively was 6 +/- 5 days, which reflects our policy that initially well-nourished patients who are experiencing a systemic inflammatory response should not undergo more than 5 to 7 days of inadequate feeding. The duration of TPN overall was 11 +/- 10 days, which primarily illustrates the dramatic reduction in length of hospital stay that has occurred throughout the health care system and the willingness to provide TPN in alternative settings including transitional care units, rehabilitation hospitals, and for short-term care, the patient's home. The most common specific reasons identified for initiating TPN rather than enteral nutrition were ileus (25%), an underlying acid-base or electrolyte/mineral disorder (13%) requiring correction, and the convenience of TPN because a central venous catheter was in place (12%). The usual indication for nutritional support at this tertiary referral and specialty hospital was actual or impending protein calorie malnutrition. TPN was chosen for a variety of reasons related to actual or anticipated tolerance to enteral feeding. This audit demonstrates that our TPN practice has evolved in relation to time of initiation and duration of feeding, which reflect a clearer appreciation of the risks and benefits of TPN. PMID- 15539244 TI - Enhancement of sucrase-isomaltase gene expression induced by luminally administered fructose in rat jejunum. AB - We have previously shown that feeding a diet containing sucrose to rats causes an elevation of sucrase-isomaltase (SI) mRNA level in the jejunum. In this study, we examined whether the SI mRNA level could be directly elevated by administration of one of the constituting monosaccharides (i.e., glucose and/or fructose). Gastric intubation of a sucrose solution caused increases in both sucrase activity and SI mRNA level in the jejunum. Intrajejunal intubation of fructose, but not glucose, led to an elevation of sucrase activity and SI mRNA level. To examine whether fructose directly affects the gene expression of SI at the segment where the absorption of this sugar takes place or the sugar-induced increase in the gene expression of SI is secondary to any possible changes in the level(s) of certain hormonal factor(s) in the blood stream, a solution containing either fructose or glucose was simultaneously perfused into two consecutive cannulated and irrigated loops of jejunum that were not isolated from blood circulation. Compared with the loop perfused with glucose, the loop perfused with fructose exhibited significantly greater sucrase activity and SI mRNA level as well as the elevated GLUT5 mRNA level. These results suggest that fructose is capable of directly increasing the gene expression of SI and GLUT5 in the confined segment where fructose is absorbed. PMID- 15539245 TI - In vitro digestibility and fermentability of levan and its hypocholesterolemic effects in rats. AB - This study describes the in vitro digestibility and fermentability of high molecular weight (ca. 2,000,000) levan and its effect on the metabolism of lipids in growing rats fed cholesterol-free diets. Levan was synthesized from sucrose using bacterial levansucrase immobilized on a honeycomb-shaped ceramic support. Although body weight gain, weight of visceral organs, morphologic changes in the digestive tract, and the serum triacylglycerol and glucose concentrations were not affected by feeding levan diets for 4 weeks, a significant hypocholesterolemic effect was observed. Serum cholesterol level was decreased to 83% or 59% by feeding a 1% or 5% levan diet, respectively. The hypocholesterolemic effect was accompanied by a significant increase in fecal excretion of sterols and lipids. High molecular weight levan, though not hydrolyzed by the salivary amylases, was hydrolyzed by artificial gastric juice and was changed to a low molecular weight (ca. 4,000) levan with a small amount of fructose, but did not produce any fructooligosaccharides. Low molecular weight (ca. 6,000) levan was not hydrolyzed by either pancreatic juice or small intestinal enzymes. This suggests that, in vivo, low molecular weight levan derived from the high molecular weight material is not further digested and reaches the colon intact. The fermentation of low molecular weight levan (ca. 6,000) by several strains of bifidobacteria was not observed. These results showed that the hypocholesterolemic effect of levan may result from the prevention of intestinal sterol absorption, and not from the action of the fermentation products of levan. PMID- 15539246 TI - R-(+)-perillyl alcohol-induced cell cycle changes, altered actin cytoskeleton, and decreased ras and p34(cdc2) expression in colonic adenocarcinoma SW480 cells. AB - Monoterpenes as S-(-)-perillyl alcohol (PA) have been shown to inhibit the isoprenylation of such growth regulatory proteins as ras. In this study, we investigated the effects of the R-(+) enantiomer of PA on cell cycle, signaling, and cytoskeletal control in the colonic adenocarcinoma cell line SW480, which carries a K-ras mutation. Cell cycle analysis by flow cytometry of SW480 cells treated with 1 mM PA for 24 hours demonstrated an increase in the number of cells in G0/G1 with a decrease in S phase, compared with untreated control cells. These cell cycle changes correlated with an inhibition of protein isoprenylation from (14)C-mevalonate and decreased expression of the cell cycle regulatory kinase p34(cdc2). Additionally, PA-treated cells acquired a flattened morphology with a condensation of cytoskeletal actin spikes to the periphery. This was in contrast to treatment with 15 microM mevinolin (MVN), a direct mevalonate synthesis inhibitor, which imparted to SW480 cells a more rounded and spindly morphology, associated with the depolymerization of actin microfilaments. Together, these data suggest that fluctuations in mevalonate and isoprenoid pools may involve different morphologic phenomenon. Because ras mediated signaling is related to the organization of the actin cytoskeleton, we investigated the effects of PA on the isoprenylation of ras. Although MVN treatment inhibited ras farnesylation, PA treatment decreased the expression of total ras protein. In summary, R-(+)-PA induced cell signaling events correlated with alterations in the organization of cytoskeletal actin and decreased protein expression of growth regulatory proteins, such as ras and cdc2 kinase. These effects may contribute to the growth inhibitory activity of R-(+)-PA. PMID- 15539247 TI - Further studies of diabetes-prone BHE/Cdb rats: increased sensitivity to calcium ion suppression of oxidative phosphorylation. AB - The responsiveness of hepatic mitochondria isolated from hyperthyroid and control Sprague-Dawley (SD) and diabetes-prone BHE/Cdb rats was studied. Hyperthyroidism was induced through the addition of thyroxine (T(4)) to the diet (2 mg/kg of diet). Oxidative phosphorylation (OXPHOS) with the addition of adenosine diphosphate (ADP) or an 8:1 mixture of adenosine monophosphate (AMP):ADP was studied. Dose response curves of state 3 and state 4 respiration, respiratory control (RC) ratio, and ADP:O ratio to calcium levels (0-7.5 microm) were generated. Mitochondria from BHE/Cdb rats were more sensitive to the addition of calcium than mitochondria from SD rats, as judged by losses in OXPHOS. T(4) treatment potentiated this strain difference and we conclude that the diabetes phenotype in the BHE/Cdb rat is probably related not only to the previously described mutation in the F(O)ATPase but also to a defect in the efflux of the calcium ion that, in turn, affected the regulation of OXPHOS. PMID- 15539248 TI - Reduced insulin secretion in response to nutrients in islets from malnourished young rats is associated with a diminished calcium uptake. AB - Changes in (45)Ca uptake and insulin secretion in response to glucose, leucine, and arginine were measured in isolated islets derived from 4-week-old rats born of mothers maintained with normal protein (NP, 17%) or low protein (LP, 6%) diet during pregnancy and lactation. Glucose provoked a dose-dependent stimulation of insulin secretion in both groups of islets, with basal (2.8 mmol/L glucose) and maximal release (27.7 mmol/L glucose) significantly reduced in LP compared with NP islets. In the LP group the concentration-response curve to glucose was shifted to the right compared with the NP group, with the half-maximal response occurring at 16.9 and 13.3 mmol/L glucose, respectively. In LP islets, glucose induced first and second phases of insulin secretions were drastically reduced. In addition, insulin response to individual amino acids, or in association with glucose, was also significantly reduced in the LP group compared with NP islets. Finally, in LP islets the (45)Ca uptake after 5 minutes or 90 minutes of incubation (which reflect mainly the entry and retention, respectively, of Ca(2+)), was lower than in NP islets. These data indicate that in malnourished rats both initial and sustained phases of insulin secretion in response to glucose were reduced. This poor secretory response to nutrients seems to be the consequence of an altered Ca(2+) handling by malnourished islet cells. PMID- 15539249 TI - Effect of maternal diet on the distribution of phospholipids and their fatty acid composition in Xenopus laevis embryos. AB - We determined the total phospholipid content, the percentage distribution of different phospholipid classes and their fatty acid composition in 6-day-old embryos obtained from Xenopus laevis females fed on two different diets. A first group of females was fed on beef liver, and a second one was nourished with commercial fish food very rich in omega-3 fatty acids. The embryos showed different patterns of phospholipids that had dissimilar fatty acid compositions. Phosphatidylinositol content was particularly affected. Due to the functional roles of this phospholipid as part of the transmembrane signaling machinery, it is possible to hypothesize that maternal diet might influence cell metabolism in amphibian embryos. PMID- 15539250 TI - Proabsorptive effect of glycerol as a glucose substitute in oral rehydration solutions. AB - We hypothesized that glycerol, a readily diffusable hydrophilic substance, may effectively substitute for glucose and enhance intestinal water and sodium absorption in an oral rehydration solution (ORS). This was evaluated using a low osmolality (230-240 mOsm/kg) ORS containing 75 mmol/L sodium and a combination of glucose:glycerol (in mmol/L) 75:0, 50:25; 37.5:37.5, 25:50, 10:65, or 0:75 during 3-hour long in vivo rat jejunal perfusions. Water, sodium, potassium, glucose and glycerol absorption, and unidirectional fluid movement (J(in), J(eff)) were determined. Sodium and net water absorptions were maximal at glucose:glycerol ratios between 37.5:37.5 and 10:65 mmol/L. In the absence of glucose (0:75), absorption of water and electrolytes was lower than at any other concentration. The greater net rehydration seemed to be due to a higher J(in) as glycerol was increased up to 65 mmol/L. Potassium absorption followed a similar pattern. With 50 mmol/L glycerol and 25 mmol/L glucose, there was a marked expansion of the lamina propria extracellular space and increased intercellular expansion between enterocytes. These results indicate that glycerol may be an effective partial substitute for glucose in ready-to-use ORS by producing an improved rate of water and electrolyte absorption. PMID- 15539251 TI - Comparison of two methods for the measurement of rat liver methylmalonyl-coenzyme A mutase activity: HPLC and radioisotopic assays. AB - Methylmalonyl-coenzyme A mutase (MCM) is a 5'-deoxyadenosylcobalamin-linked mitochondrial enzyme that catalyzes the isomerization of L-methylmalonyl-coenzyme A to succinyl-coenzyme A. In vitro assays of total and holo-MCM activities are important tools for investigating the cobalamin pathway. Several methods have been described for measuring MCM activity. The most commonly-used method is a radioassay based on the permanganate oxidation of DL[CH(3)-(14)C]methylmalonyl coenzyme A, but radiometric methods are insensitive, laborious, and time consuming. Therefore, we have compared this method with a nonradiometric assay, potentially most sensitive, based on the separation of methylmalonyl-coenzyme A and succinyl-coenzyme A by high performance liquid chromatography (HPLC). We determined the optimal assay conditions and the reproducibility and sensitivity of each technique. The results obtained by the two techniques were very different: the specific activities obtained by the permanganate oxidation method (0.039 +/- 0.013 nmol/min/mg protein for the holo-MCM activity and 1.90 +/- 0.69 nmol/min/mg protein for the total-MCM activity) were threefold lower than those obtained with the HPLC method (0.124 +/- 0.011 nmol/min/mg protein for the holo MCM activity and 6.15 +/- 0.76 nmol/min/mg protein for the total-MCM activity). The coefficients of variation for the radiometric method (18.4-40.6%) were three to five times greater than those for the HPLC assay (3.5-12.2%). This demonstrates the lack of sensibility and reproducibility of the permanganate radioassay. Thus, the radiometric method is not suitable for measuring low mutase activities such as the holo activities in tissues. The intrinsic inconvenience of the radiometric assay indicates that the HPLC method is a method of choice for measuring MCM activity. PMID- 15539252 TI - Effect of dietary selenium restriction on selected parameters of selenium status in men with high life-long intake. AB - The influence of selenium (Se) restriction on disposition in plasma and urine fractions of infused (74)Se (selenite) was studied when adult males (Enshi City, Hubei Province, PRC) whose habitual daily Se intake is approximately 480 microg per day were transferred to Lichuan County, where the daily intake is approximately 30 microg. The subjects received an infusion (106 microg Se) on the day before consuming foods low in Se and a second infusion (113 microg Se) 63 days later. Blood and 24-hour urine samples were collected each day for 7 days after the first infusion and on days 22, 43, and 62 following the first infusion. Urine and blood were also collected daily for the next 7 days after the second infusion. Plasma total Se concentration increased for 7 days after each of the two infusions and urine Se decreased exponentially following both the first and second infusions. The excretion of trimethylselenonium followed the same pattern as the total urinary Se. Surprisingly, there was not a significant difference in selenite retention between the two infusion periods, and the data indicated that, regardless of the chemical form of Se present in various organs, its catabolism leading to excretion in urine followed the same pathway as that of selenite. Labeled Se was incorporated predominantly in the plasma selenoprotein P fraction and the half-life of Se in this fraction was determined to be 1.9 to 2.9 days. Thus, a longer depletion period is required in these subjects to obtain more significant changes. PMID- 15539253 TI - Accumulation and retention of micellar beta-carotene and lutein by Caco-2 human intestinal cells. AB - Despite the interest in the diverse roles of dietary carotenoids in human health, little is known about the transfer of these plant pigments from foods to micelles during digestion and their subsequent transfer across the intestinal epithelium. We conducted this study to characterize the intestinal uptake of micellarized carotenoids using monolayers of differentiated Caco-2 human intestinal cells. Crystalline beta-carotene (BC) and lutein (LUT), solubilized in mixed micelles for delivery to cells, were stable in a tissue culture environment for 20 hours. Cellular accumulation of micellar BC and LUT was proportional to the media content of carotenoids at /=18 micromol/L. There was no indication that high levels of BC in medium or within cells adversely affected micellar LUT accumulation. These data support the use of the Caco-2 human cell line as a model for studying the intestinal uptake, absorption, and possible interactions of dietary carotenoids. PMID- 15539254 TI - Effects of dietary omega3 and omega6 lipids and vitamin E on proliferative response, lymphoid cell subsets, production of cytokines by spleen cells, and splenic protein levels for cytokines and oncogenes in MRL/MpJ-lpr/lpr mice. AB - omega3 Fatty acid rich fish oil (FO) and vitamin E may delay the progress of certain autoimmune diseases. The present study examined the mechanisms of action of omega3 lipids and vitamin E in autoimmune-prone MRL/lpr mice suffering from extensive lymphoproliferation, lupus-like symptoms, and accelerated aging. To determine whether the effects of omega3 lipids in autoimmune disease is linked to vitamin E levels, weanling female MRL/lpr and congenic control MRL/++ mice were fed diets containing 10% corn oil (CO) or 10% FO at two levels of vitamin E (75 IU or 500 IU/kg diet) for 4 months. The appearance of lymph nodes was delayed in the mice fed FO, and higher levels of FO offered further protection against the appearance of lymph nodes. Analysis of the spleen cells revealed that the cells positive for Thy.1 and Fas were significantly higher in the MRL/++ mice. The groups fed high levels of vitamin E generally exhibited higher levels of Fas. The proliferative response of splenocytes of MRL/++ mice to mitogens was significantly higher compared with MRL/lpr mice. Interleukin (IL)-10 production by spleen cells was significantly higher in FO-fed MRL/lpr mice than in CO-fed mice. In mice fed a high level of vitamin E, the production of IL-12 and tumor necrosis factor-alpha was significantly lower and IL-2 was significantly higher than in animals fed a low level of vitamin E. Proinflammatory cytokines were higher in the MRL/lpr mice and both FO and vitamin E lowered the levels of proinflammatory cytokines and lipid mediators. Western blots revealed that c-myc and c-ras were significantly lower and IL-2 and transforming growth factor (TGF) beta1 levels were significantly higher in the spleens of MRL/++ mice. FO lowered c-myc and high levels of vitamin E in the diets normalized the levels of TGF beta1 in MRL/lpr mice. The observations from this study suggest that both FO and vitamin E modulate the levels of specific cytokines, decrease the levels of proinflammatory cytokines, inflammatory lipid mediators, and c-myc, and increase TGF-beta1 levels in spleens of MRL/lpr mice and thus may delay the progress of autoimmune diseases. PMID- 15539255 TI - Effects of a prudent diet containing either lean beef and mutton or fish and skinless chicken on the plasma lipoproteins and fatty acid composition of triacylglycerol and cholesteryl ester of hypercholesterolemic subjects. AB - In this two-phase crossover study, 39 hypercholesterolemic subjects followed a prudent diet with either lean red meat or fish and skinless chicken (treatment groups), and 13 subjects (reference group) followed their habitual diet. Fasting blood samples were analyzed for plasma total cholesterol, triacylglycerol (TAG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein one- and two-cholesterol, apolipoprotein-B, very low density lipoprotein cholesterol, and very low density lipoprotein TAG, and fatty acid composition of plasma TAG and cholesteryl ester (CE). Body mass and blood pressure were determined. Seven-day dietary records were kept once at baseline and twice during the treatment periods. Significant differences were observed in dietary intake between the baseline and treatment diets and between the two treatment diets. HDL-C (P < 0.05) and diastolic blood pressure (P < 0.01) were higher in patients on the red meat diet than in those on the chicken-fish diet. No other significant differences in lipoproteins were observed between the effects of the two treatment diets. The linoleic acid (%), eicosapentaenoic acid (%), and the eicosapentaenoic acid/arachidonic acid ratios in TAG and CE were higher (P < 0.01) in subjects on the chicken-fish diet than in those on the red meat diet. In conclusion, this study showed that the effect of two lipid-lowering diets containing either lean red meat or skinless chicken and fish on the atherogenic lipoproteins did not differ significantly. A prudent diet with skinless chicken and fish, however, had a more favorable effect on the fatty acid composition of the plasma TAG and the CE than did the lean red meat diet. PMID- 15539256 TI - Effects of fasting on lipoprotein lipase activity in different depots of white and brown adipose tissues in diet-induced overweight rats. AB - The aim of the present study was to evaluate the effects of 24 hours of starvation on lipoprotein lipase (LPL) activity in various depots of white and brown adipose tissues in control rats and in rats with two different degrees of overweight, both induced by dietary treatment. In control rats, no changes in LPL immunoreactive mass were observed in either white or brown adipose tissues after fasting, whereas the effects of food deprivation on enzyme activity were opposite in white versus brown adipose tissues. The LPL activity response to fasting was impaired by obesity: White adipose depots of cafeteria obese rats showed a lower ability to downregulate LPL during fasting and the increased LPL activity induced by fasting in brown adipose depots was less intense in the obese rats compared with control animals. When the degree of overweight was reduced, the differences between obese and control rats were also attenuated. PMID- 15539257 TI - Malnutrition sequela on the drug metabolizing enzymes in male Holtzman rats. AB - The effect of food restriction on the specific activities of the drug metabolizing enzymes (DME) system was studied in Holtzman male rats by comparing DME activities in 90-day-old control rats fed ad libitum (CO), rats fed 40% restricted food (RF) from the gestation period to the day of sacrifice, and recovered rats (rRF) fed 40% restricted food from period of gestation to 45 days of age and then fed ad libitum until the day of sacrifice. In liver, total cytochrome P450 (CYP) of the RF and rRF groups was higher by approximately 50% and 28%, respectively, than in CO rats. Specific activities of individual CYP monooxygenases (MO) such as CYP2B [7-methoxycoumarin demethylase (MOCD)], CYP1A [aryl hydrocarbon hydroxylase (AHH) and ethoxyresorufin deethylase (EORD)], and CYP2E [nitrosodimethylamine demethylase (NDMAd)] were 31, 61, 43, and 56% in RF and 16, 36, 26, and 32% in rRF groups, respectively, more than the CO values. Conjugases such as UDP- glucuronosyltransferases with substrates 3-OH benzo(a)pyrene (UGT1) and 4-hydroxybiphenyl (UGT2) and glutathione S-transferase (GST) with substrate 1-chloro-2,4-dinitrobenzene were higher by 72, 69, and 33% in RF and 28, 38, and 24% in rRF groups, respectively. MO activities (MOCD and EORD) were significantly higher in lung, kidney, and intestine: MOCD by 82, 48, and 45% in RF and 40, 25, and 22% in rRF, respectively; and EORD by 84, 77, and 67% in RF and 40, 33, and 28% in rRF, respectively. However, activity of conjugases (UGT1 and GST) were significantly lower (approximately 35-45%) in RF and rRF rats (approximately 20-30%) than in the CO group in above mentioned extrahepatic tissues. These studies indicate that undernourishment during the period of gestation, weanling, and growth and development of microsomal enzymes produces a sequela of events on the DME in hepatic and extrahepatic tissues that cannot return to the control values even when fed ad libitum. PMID- 15539258 TI - Effects of dietary supplementation with sea buckthorn (Hippophae rhamnoides) seed and pulp oils on atopic dermatitis. AB - A placebo-controlled, double-blind study was conducted to investigate the effects of seed and pulp oils of sea buckthorn (Hipphophae rhamnoides) on atopic dermatitis. Linoleic (34%), alpha-linolenic (25%), and oleic (19%) acids were the major fatty acids in the seed oil, whereas palmitic (33%), oleic (26%), and palmitoleic (25%) acids were the major fatty acids in the pulp oil. The study group included 49 atopic dermatitis patients who took 5 g (10 capsules) of seed oil, pulp oil, or paraffin oil daily for 4 months. During follow-up dermatitis improved significantly in the pulp oil (P < 0.01) and paraffin oil (P < 0.001) groups, but improvement in the seed oil group was not significant (P = 0.11). Supplementation of seed oil increased the proportion of alpha-linolenic acid in plasma neutral lipids (P < 0.01), and increases of linoleic, alpha-linolenic, and eicosapentaenoic acids in plasma phospholipids were close to significant (0.05 < P < 0.1). Pulp oil treatment increased the proportion of palmitoleic acid (P < 0.05) and lowered the percentage of pentadecanoic acid (P < 0.01) in both plasma phospholipids and neutral lipids. In the seed oil group, after 1 month of supplementation, positive correlations were found between symptom improvement and the increase in proportions of alpha-linolenic acid in plasma phospholipids (Rs = 0.84; P = 0.001) and neutral lipids (Rs = 0.68; P = 0.02). No changes in the levels of triacylglycerols, serum total, or specific immunoglobulin E were detected. In the pulp oil group, a significant (P < 0.05) increase in the level of high density lipoprotein cholesterol, from 1.38 to 1.53 mmol/L was observed. PMID- 15539259 TI - Effects of isoflavones containing soy protein isolate compared with fish protein on serum lipids and susceptibility of low density lipoprotein and liver lipids to in vitro oxidation in hamsters. AB - The effects of dietary soy protein isolate (SPI), ethanol-extracted SPI (E-SPI) low in isoflavones, and fish protein (FP) on the concentration of blood lipids and the susceptibility of low density lipoprotein (LDL) to copper-induced oxidation were compared in male golden Syrian hamsters fed a moderate hypercholesterolemic semi-purified diet for 10 weeks. SPI, E-SPI, and FP were incorporated into the isonitrogenous experimental diets as protein sources. The SPI group exhibited significantly lower serum total cholesterol concentration compared with the E-SPI group (P < 0.05) and the FP group (P < 0.01). Both the SPI and E-SPI groups showed lower LDL cholesterol (P < 0.001 and P < 0.05, respectively) and less LDL apolipoprotein B (P < 0.01) compared with the FP group. The distribution pattern of serum lipoprotein cholesterol fractions of the SPI and E-SPI groups were similar to each other, but different from that of the FP group. The lysine/arginine ratio of the three diets was significantly correlated with serum total cholesterol concentration (r = 0.462, P = 0.023). The resistance of LDL to copper-induced oxidation was greater in the SPI group than in the E-SPI and FP groups as assessed by the lower concentrations of thiobarbituric acid-reactive substances (TBARS) and the longer lag time required for the formation of conjugated dienes (P < 0.01). Livers of hamsters fed the FP diet had a higher amount of TBARS than those of hamsters fed SPI (P < 0.01) and E SPI (P < 0.05) diets. The SPI diet showed sparing effects on alpha-tocopherol contents in both serum and liver. It seems likely that soy isoflavones protect the circulating and membrane lipids by sparing alpha-tocopherol and endogenous antioxidants. PMID- 15539260 TI - Effect of delta22-5beta-taurocholenic acid on hepatic cholesterol and fatty acid in gold thioglucose obese mice fed low- or high-fat diets. AB - We previously demonstrated that hyperglycemic-obese (obob) mice fed a 1% corn oil diet accumulated 10 times as much hepatic cholesterol as did their non-obese (+/?) littermates fed this diet because of difficulty in removal of cholesterol from the liver rather than from increased synthesis. Furthermore, feeding the bile acid analog Delta(22)-5beta-taurocholenic acid completely prevented the accumulation of hepatic cholesterol in obob mice fed the 1% corn oil diet. The hypothesis to be tested in the current study is that these aspects of cholesterol metabolism in the obob mouse do not occur in the hyperinsulinemic and insulin resistant gold thioglucose obese mouse. Gold thioglucose obese (gtgo) and non obese (ngtgo) mice were fed diets containing either 1% corn oil or 40% lard each with or without added taurocholenic acid for 6 weeks and then given a 250 mg meal of [U-(14)C]-glucose with incorporation of label into hepatic cholesterol and fatty acid measured 2 hours later. Consistent with earlier results in the obob model, incorporation of labeled glucose was significantly increased in obese compared with non-obese mice fed 1% corn oil and significantly reduced either by feeding 40% lard or by adding taurocholenic acid to the diet. In addition, taurocholenic acid greatly increased incorporation of labeled glucose into hepatic cholesterol in obese or non-obese mice fed either diet. In contrast to obob mice, the percentage of fat in the liver of gtgo mice was increased only 50% compared with ngtgo mice. The comparable increase in obob mice was 480%. Hepatic cholesterol did not increase significantly in the liver of gtgo mice fed 1% corn oil when compared with the ngtgo controls. The comparable increase in obob mice fed 1% corn oil was 350%. Also in marked contrast to obob mice, feeding taurocholenic acid increased hepatic cholesterol compared with non-obese controls fed either diet. The results are discussed in the light of the presence of circulating leptin in gtgo but not in obob mice. PMID- 15539261 TI - Glucocorticoid-dependent induction of HMG-CoA reductase and malic enzyme gene expression by polychlorinated biphenyls in rat hepatocytes. AB - Administration of xenobiotics to rats results in hypercholesterolemia and in the induction of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and malic enzyme. To investigate the mechanism of the induction of the enzymes by xenobiotics, the effects of xenobiotics on gene expressions for HMG-CoA reductase, malic enzyme, and cytochrome P-450 in rat liver and in cultured hepatocyte were investigated. The treatment of rats with polychlorinated biphenyls (PCB) as a xenobiotic induced mRNAs for HMG-CoA reductase and malic enzyme as well as CYP2B1/2 (cytochrome P-450b/e). Other xenobiotics, 1,1,1 trichloro-2,2-bis(p-chlorophenyl)ethane (DDT), and chloretone, also increased HMG CoA reductase mRNA. In an investigation of diurnal rhythm of mRNA for HMG-CoA reductase, the induction by PCB was observed in a dark period. Induced expressions of HMG-CoA reductase gene and malic enzyme gene by PCB were observed in primary cultured rat hepatocytes and showed that the action of PCB on the gene expression relating to lipid metabolism was directed on hepatocytes. The induction was observed only in hepatocytes cultured on Engelbreth-Holm-Swarm sarcoma basement membrane gel (EHS-gel), not on type I collagen, which is usually used for monolayer culture of hepatocytes. The induction of CYP2B1/2 gene expression also was observed only in the cells cultured on EHS-gel. The induction of HMG-CoA reductase and malic enzyme by PCB required dexamethasone. However, the addition of dexamethasone per se to medium containing insulin did not show an inductive effect on levels of mRNA for HMG-CoA reductase and malic enzyme. From the data of diurnal variation and hepatocyte culture experiment, HMG-CoA reductase and malic enzyme are considered to be induced by PCB through the so called "permissive effect" of glucocorticoid. PMID- 15539262 TI - Allyl mercaptan, a major metabolite of garlic compounds, reduces cellular cholesterol synthesis and its secretion in Hep-G2 cells. AB - The cytotoxicity, cellular cholesterol synthesis, and secretion of allyl mercaptan, a major metabolite of garlic compounds, were studied in Hep-G2 cells. The cells were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum and treated with 5, 25, 50, 100, 125, 250, and 500 microg of allyl mercaptan/mL for 4, 12, and 24 hours. At concentrations up to 125 microg, no significant cytotoxic effect was noted during those incubation periods. However, at a concentration of 250 microg, cell viability decreased approximately 50% compared with the control (P < 0.05) in all three incubation times. At a concentration of 500 microg, allyl mercaptan was highly toxic, causing extensive cell death. The treatment of cells with 5, 10, 25, 50, or 100 microg (noncytotoxic concentration) of allyl mercaptan resulted in a marked inhibition of (3)H-acetate incorporation into cholesterol. At concentrations between 5 and 100 microg, the cholesterol synthesis was inhibited 20 to 80% in cells and the cholesterol secretion into the medium decreased 20 to 50% compared with the control (P < 0.05). The concentration of allyl mercaptan required to suppress cholesterol synthesis by 50% was approximately 25 microg/mL. Allyl mercaptan treatment of cells incubated with 1 mM of oleic acid also resulted in a significant decrease in the cholesterol synthesis compared with the cells incubated with oleic acid alone (19.5 +/- 1.2 x 10(3) dpm/mg protein/4 h vs. 30.0 +/- 2.6 x 10(3) dpm/mg protein/4 h; P < 0.05). The present study demonstrates that allyl mercaptan is effective in inhibiting cholesterol synthesis at concentrations as low as 5 microg, and its inhibition is concentration dependent. PMID- 15539263 TI - Ovariectomized hamster: a potential model of postmenopausal hypercholesterolemia. AB - A suitable and economical animal model of ovarian hormone deficiency can greatly enhance the understanding of postmenopausal-elevated risk of coronary heart disease. The male Golden Syrian hamster is a well-established small animal model of hypercholesterolemia, but the effect of ovariectomy on lipid profile in the female hamster is unclear. The objective of this study was to determine whether ovariectomized hamsters develop hypercholesterolemia and experience changes in body fat distribution consistent with changes observed in postmenopausal women. Twenty-two 90-day-old female Golden Syrian hamsters were divided into two groups and were either ovariectomized or sham-operated and given free access to a standard cholesterol-free laboratory diet for 65 days. Ovariectomized hamsters had significantly (P < 0.05) elevated serum total cholesterol concentrations (16.6%) as well as abdominal fat mass (56%; P< 0.01) despite equal food intake compared with the sham-operated group. In contrast, the mean intestinal weight and in vivo rate of sterol biosynthesis were significantly (P < 0.002 and P = 0.01, respectively) lower in the ovariectomized compared with the sham-operated group. In vivo rates of hepatic sterol biosynthesis were directionally lower (P = 0.1) in the ovariectomized group. No significant differences were observed in final body weight, serum triglycerides, or liver total cholesterol and lipids between the two groups. In conclusion, ovariectomized hamsters undergo changes in serum cholesterol and fat distribution similar to those experienced by postmenopausal women, and thus may serve as an appropriate model for postmenopausal hypercholesterolemia. PMID- 15539264 TI - Effects of phytoestrogen-coumestrol on lipid and carbohydrate metabolism in young ovariectomized rats may be independent of its estrogenicity. AB - Two groups of young ovariectomized female rats received one of two treatments. The first group was fed coumestrol in lab chow (200 microg of coumestrol per day) for 14 days; the second group received coumestrol (40 mg/L) via perfusion medium. There was a significant increase (78% compared with the control group) in the uterine weight after coumestrol treatment, which supports the estrogen-like activity of coumestrol. Phytoestrogen diminished the liver and skeletal muscle glycogen contents by 18% and 29%, respectively, and increased the blood glucose level by 24%. Glycogenolytic activity of coumestrol was observed when it acted directly on the liver areas. Although phytoestrogen did not influence insulin and glucagon blood level, liver and to some degree muscle susceptibility to insulin (measured as hormone binding by insulin receptors) was decreased. Coumestrol increased the content of triglycerides in muscle by 113% and enhanced the liver lipid synthesis from glucose by 179%. Liver cholesterol concentration was increased both after coumestrol feeding (by 12%) and when it acted directly on the liver (by 16%). These observations suggest that coumestrol is in general anabolic with regard to lipid and catabolic within-carbohydrate metabolism of young ovariectomized female rats. Based on the results of this study, it is concluded that influence of coumestrol on lipid and carbohydrate metabolism of ovariectomized rats is in part not related to its estrogenic action. PMID- 15539265 TI - Cefetamet pivoxil treatment causes loss of carnitine reserves that can be prevented by exogenous carnitine administration. AB - Two groups of pediatric patients receiving cefetamet pivoxil treatment (3 x 500 mg daily) for 7 days were studied. In the first group (Group A) the drug was administered alone; in the second group (Group B) the drug was given in combination with a molar excess of carnitine (3 x 1 g). Medication with cefetamet pivoxil alone was associated with a large urinary excretion of pivaloylcarnitine: Approximately 71% of the daily pivalate intake could be eliminated as carnitine ester in the urine. In this group, the plasma level and the urinary output of free carnitine decreased. By contrast, in Group B, the administration of molar excess of carnitine aided stochiometric elimination of pivalate as carnitine ester, and the plasma levels and carnitine-free urinary output were unchanged. The data show that medication of cefetamet pivoxil results in the formation of pivaloylcarnitine in children; the sustained loss of carnitine esters can ultimately lead to carnitine deficiency. Molar excess of exogenous carnitine aids in the elimination of pivalate derived from cefetamet pivoxil therapy and helps to maintain the carnitine reserves. PMID- 15539266 TI - Oxidative stress and antioxidant status in mouse kidney: effects of dietary lipid and vitamin E plus iron. AB - The purpose of this study was to determine the effects of dietary fat, vitamin E, and iron on oxidative damage and antioxidant status in kidneys of mice. Sixty 1 month-old male Swiss-Webster mice were fed a basal vitamin E-deficient diet that contained either 8% fish oil + 2% corn oil or 10% lard with or without 1 g all rac-alpha-tocopherol acetate or 0.74 g ferric citrate per kilogram of diet for 4 weeks. Significantly (P < 0.05) higher levels of lipid peroxidation products, thiobarbituric acid reactants (TBAR), and conjugated dienes were found in the kidneys of mice fed with fish oil compared with mice fed lard irrespective of vitamin E status. Mice maintained on a vitamin E-deficient diet had significantly higher renal levels of TBAR, but not conjugated dienes, than the supplemented group. Fish oil fed mice receiving vitamin E supplementation had lower levels of alpha-tocopherol than did mice in the lard fed group. Significantly higher levels of ascorbic acid were also found in the kidneys of mice fed with fish oil than were found in mice fed lard. The levels of protein carbonyls and glutathione (GSH), and activities of catalase, superoxide dismutase, selenium (Se)-GSH peroxidase, and non-Se-GSH peroxidase were not significantly altered by dietary fat or vitamin E. Dietary iron had no significant effect on any of the oxidative stress and antioxidant indices measured. The results obtained provide experimental evidence for the pro-oxidant effect of high fish oil intake in mouse kidney and suggest that dietary lipids play a key role in determining cellular susceptibility to oxidative stress. PMID- 15539267 TI - Transgenic animals and nutrition research. AB - Transgenic animals are useful tools for the study of biological functions of proteins and secondary gene products synthesized by the action of protein catalysts. Research in nutrition and allied fields is benefiting from their use as models to contrast normal and altered metabolism. Although food, nutritional products, and ingredients from transgenic animals have not yet reached consumers, the technologies for their production are maturing and yielding exciting results in experimental and farm animals. Regulatory governmental bodies are already issuing guidelines and legislation in anticipation of the advent of these products and ingredients. This review summarizes available technology for the production of transgenic animals, discusses their scientific and commercial potential, and examines ancillary issues relevant to the field of nutrition. PMID- 15539268 TI - Tissue carnitine fluxes in vitamin C depleted-repleted guinea pigs. AB - The biosynthesis of carnitine requires vitamin C as a cofactor for two separate hydroxylation steps. The majority of body carnitine (approximately 98%) is located in muscle and less than 0.5% is present in plasma. We examined the physiologic dynamics of plasma free carnitine and muscle total acid-soluble carnitine in vitamin C-depleted guinea pigs repleted with increasing amounts of vitamin C. Animals were fed a vitamin C-deficient diet for 3 weeks at which time symptoms of scurvy were evident. Animals were repleted with increasing doses of vitamin C, from 0.5 to 10.0 mg vitamin C/100 g body weight daily. Muscle total acid-soluble carnitine concentrations tended to correlate directly with plasma vitamin C (r = 0.41, P = 0.087) during the repletion phase of the study. Conversely, plasma free carnitine was inversely related to liver vitamin C (r = 0.54, P = 0.020) and to muscle total acid-soluble carnitine (r = -0.56, P = 0.015). Mean plasma free carnitine values fell 30% over the course of vitamin C repletion (P > 0.05) and mean muscle total acid-soluble carnitine rose by 30% (P > 0.05). These data suggest that elevated plasma free carnitine may indicate a low to marginal vitamin C status. PMID- 15539269 TI - Effects of dietary polyamines and clofibrate on metabolism of polyamines in the rat. AB - The activities of catalase, polyamine oxidase, diamine oxidase, ornithine decarboxylase, and peroxisomal beta-oxidation were assayed in homogenates from liver and small intestinal mucosa of rats which had been fed either a diet very low in polyamines or a diet containing five times the levels of dietary polyamines (putrescine, spermine, and spermidine) found in a standard rat diet. In rats fed the high polyamine diet, hepatic activities of catalase and polyamine oxidase were significantly decreased. Levels of the other activities were unchanged, except that intestinal ornithine decarboxylase was decreased. In rats treated simultaneously with clofibrate, the high polyamine diet restored activities of catalase, ornithine decarboxylase, and polyamine oxidase back to levels found in rats fed the low polyamine diet. The expected increase in activity of peroxisomal beta-oxidation was observed, although this was somewhat diminished in rats fed the high polyamine diet. Intestinal diamine oxidase activity was stimulated by clofibrate, particularly in rats fed the high polyamine diet. For the duration of the experiment (20 days), levels of putrescine, spermine, and spermidine in blood remained remarkably constant irrespective of treatment, suggesting that polyamine homeostasis is essentially independent of dietary supply of polyamines. It is suggested that intestinal absorption/metabolism of polyamines is of significance in this respect. Treatment with clofibrate appeared to alter polyamine homeostasis. PMID- 15539270 TI - Dietary fat type alters glucose metabolism in isolated rat hepatocytes. AB - Dietary fat type can influence the regulation of carbohydrate metabolism in multiple tissue types. The influence of feeding high-fat (40% of kilocalories) diets containing either menhaden oil (MO) or coconut oil (CO) on hepatic glycogenolytic and gluconeogenic capacities was studied in isolated rat hepatocytes. Estimates of both glycogenolytic and gluconeogenic capacities were performed on hepatocytes isolated from fed and fasted animals, respectively. In MO-fed animals, both basal and hormone-stimulated rates of glucose production were significantly greater than those in CO-fed animals. However, both groups displayed a similar maximal increase in glucose production above basal for glucagon and epinephrine (2.3- and 1.9-fold, respectively). Basal rates of adenosine 3',5'-cyclic phosphate (cAMP) production were not different between groups whereas glucagon-stimulated cAMP production was increased twofold in the MO-fed group. In both MO and CO groups, the addition of 10 nM insulin reduced glucose production in fed animals to similar absolute rates. In animals fasted for 24 hours, gluconeogenic capacity was estimated using 10 mM pyruvate, lactate, or glycerol. Glucose production from all substrates was significantly greater in CO-fed animals. In addition to increased gluconeogenic rates, maximal phosphoenolpyruvate carboxykinase (PEPCK) activity was increased in the CO-fed group. Insulin reduced glucose production in both dietary groups, but the absolute rate of glucose production was 28% greater in the CO-fed group relative to the MO-fed group. In summary, dietary fat type can markedly influence the regulation of hepatic glucose metabolism in multiple metabolic pathways. MO feeding promoted glycogenolysis and sensitivity to insulin whereas CO feeding favored gluconeogenesis and reduced insulin sensitivity. PMID- 15539271 TI - The hepatic amino acid system A transport activity, is up-regulated in obese Zucker rats. AB - The utilization of L-alanine by liver is dependent on amino acid uptake from blood. This uptake, mainly mediated by the A transport system, may be regulated by different nutritional and physiologic conditions. The regulation of this transport system by diets with different protein content was tested in lean and obese Zucker rats. High-protein (HP) and low-protein (LP) diets led to changes in the rats' growth patterns, especially in lean animals. However, homeostasis was relatively well maintained, as seen in plasma values, in spite of the increased urea production in the HP groups and increased triacylglycerides in the LP groups. The obese animals took up L-alanine at a higher rate than the lean animals. Obesity led to the emergence of a high-affinity component (K(M) approximately 0.1-0.2 mM) in the transport system, which was not dependent on the protein content of the diet. This component has a 10-fold increase in affinity for L-alanine, but with an approximately 3- to 5-fold reduction in maximal velocity of transport. PMID- 15539272 TI - Sensitivity of beta-casein phosphopeptide-iron complex to digestive enzymes in ligated segment of rat duodenum. AB - Binding iron to the phosphorylated beta(1-25) peptide derived from beta-casein improves iron bioavailability in the rat. The aim of the present work was to learn how injected beta(1-25) and iron-beta(1-25) complex behave in the duodenum of rats using the technique of intestinal ligation in situ and reversed-phase (RP)-high performance liquid chromatography-electrospray mass spectrometry analysis of the lumen contents. The results demonstrate that beta(1-25) is sensitive to digestive enzymes including proteases/peptidases and phosphatases during duodenal transit. The lumen contents of rats perfused with iron free beta(1-25) contained all peptidic sequences derived from beta(1-25). In contrast, the phosphorylated part of beta(1-25) [i.e., beta(15-24)] was not detected in lumen of rats perfused with iron-beta(1-25) complex. PMID- 15539273 TI - Mitochondrial oxidative metabolism during respiratory infection in riboflavin deficient mice. AB - Studies in children and mice have shown that respiratory infection alters riboflavin metabolism, resulting in increased urinary loss of this vitamin. This could be due to mobilization of riboflavin from the liver to blood because liver Flavin adenine dinucleotide (FAD) levels were lowered in the mice during infection. To understand the functional implications of lowered hepatic FAD levels during respiratory infection, flavoprotein functions such as oxidative phosphorylation and beta-oxidation of the liver mitochondria were examined during infection in mice. Weanling mice were fed either riboflavin-restricted or control diet for 18 days and then injected with a sublethal dose of Klebsiella pneumoniae. During infection, the state 3 respiratory rate with palmitoyl-L carnitine and glutamate were significantly lowered (27-29%) in the riboflavin restricted group, whereas in the control group 10% reduction was observed with palmitoyl-L-carnitine as substrate. A 22% reduction in the respiratory control ratio with palmitoyl-L-carnitine as substrate was observed during infection in the riboflavin-restricted group. The beta-oxidation of palmitoyl-L-carnitine was significantly lowered (29%) in the riboflavin-restricted infected group. The results of the study suggest that the effects of infection on vital physiologic functions were more pronounced in the riboflavin-restricted mice than in the control mice. PMID- 15539274 TI - Folate and carcinogenesis: evidence, mechanisms, and implications. PMID- 15539275 TI - Nonessential amino acids are not necessary to stimulate net muscle protein synthesis in healthy volunteers. AB - The present study was performed to test the hypothesis that orally administered essential amino acids, in combination with carbohydrate, will stimulate net muscle protein synthesis in resting human muscle in vivo. Four volunteers ingested 500 mL of a solution containing 13.4 g of essential amino acids and 35 g sucrose (EAA). Blood samples were taken from femoral arterial and venous catheters over a 2-hour period following the ingestion of EAA to measure arteriovenous concentrations of amino acids across the muscle. Two muscle biopsies were taken during the study, one before administration of the drink and one approximately 2 hours after consumption of EAA. Serum insulin increased from normal physiologic levels at baseline (9.2 +/- 0.8 microU/mL) and peaked (48 +/- 7.1 microU/mL) 30 minutes after EAA ingestion. Arterial essential amino acid concentrations increased approximately 100 to 400% above basal levels between 10 and 30 minutes following drink ingestion. Net nitrogen (N) balance changed from negative (-495 +/- 128 nmol/mL) prior to consumption of EAA to a peak positive value (416 +/- 140 nmol/mL) within 10 minutes of ingestion of the drink. EAA resulted in an estimated positive net N uptake of 307.3 mg N above basal levels over the 2-hour period. Muscle amino acid concentrations were similar prior to and 2 hours following ingestion of EAA. We conclude that ingestion of a solution composed of carbohydrates to stimulate insulin release and a small amount of essential amino acids to increase amino acid availability for protein synthesis is an effective stimulator of muscle protein anabolism. PMID- 15539276 TI - Effect of dietary ghee--the anhydrous milk fat, on blood and liver lipids in rats. AB - Dairy products are important sources of dietary fat in India. Anhydrous milk fat, viz., ghee, is consumed as such in the diet and also is used for frying the dishes. Ghee contains high levels of saturated fatty acids and cholesterol, which are considered risk factors for cardiovascular diseases. In the present study, ghee, at levels ranging from 0.25 to 10%, was included in a nutritionally balanced AIN-76 diet fed to Wistar rats for a period of 8 weeks. The serum lipid profiles of these animals showed a dose dependent decrease in total cholesterol, low density lipoproteins and very low density lipoproteins cholesterol, and triglyceride levels when ghee was present at levels greater than 2.5% in the diet. Liver cholesterol and triglycerides also were decreased in these animals. When ghee was included as a sole source of fat at a 10% level, polyunsaturated fatty acids in the serum and liver lipids were reduced significantly. Similar results were observed when ghee was subjected to a higher temperature (120 degrees C) to generate cholesterol oxidation products and fed to the animals. Although cholesterol oxidation products were not accumulated in serum, significant amounts were accumulated in liver only when ghee was fed as a sole source of fat at a 10% level. This study revealed that the consumption of ghee up to a 10% level in the diet altered blood lipid profiles in such a manner as not to elevate the risk factors for cardiovascular diseases. PMID- 15539277 TI - The efflux of biotin from human peripheral blood mononuclear cells. AB - Peripheral blood mononuclear cells (PMBCs) are readily available for sampling and are a useful model for studying biotin metabolism in human cells. To better understand biotin handling by PMBCs, we investigated the mechanism(s) and kinetics of biotin efflux from PMBCs. Human PMBCs were incubated with [(3)H]biotin at 475 pmol/L to load the cells. The [(3)H]biotin-loaded cells were then harvested and incubated in [(3)H]biotin-free media for up to 20 hours. At various intervals, aliquots of the PMBC suspensions were collected and analyzed for intracellular [(3)H]biotin. [(3)H]Biotin efflux from cells at 37 degrees C was fast and triphasic; the half-lives for the three elimination phases were 0.2 +/- 0.02 hours, 1.2 +/- 0.1 hours, and 21.9 +/- 13.6 hours. Such a triphasic [(3)H]biotin efflux could reflect (1) rapid efflux of free biotin, (2) slower release of biotin bound to intracellular molecules, and (3) even slower release from carboxylases in cellular organelles. Incubation at 4 degrees C rather than 37 degrees C increased the [(3)H]biotin retained at 20 hours from 27% to 85%. This observation is consistent with transporter-mediated efflux. When cellular glucose utilization was reduced by 2-deoxy-d-glucose and sodium fluoride, [(3)H]biotin efflux was similar to controls, suggesting that biotin efflux does not directly require metabolic energy. When [(3)H]biotin-loaded cells were incubated in external medium containing unlabeled biotin analogs, [(3)H]biotin efflux was accelerated approximately two times compared with incubation in a biotin-free medium. This observation suggests that biotin efflux is mediated by the same transporter that mediates biotin uptake from the extracellular medium (i.e., classic countertransport). PMID- 15539278 TI - Estradiol replacement in ovariectomized rats increases the hepatic concentration and biliary secretion of alpha-tocopherol and polyunsaturated fatty acids. AB - Previously, we showed that estradiol replacement in ovariectomized rats produced prominent increases in serum and liver alpha-tocopherol (alphaTP). The present study was conducted to examine whether the estrogen-induced increase in the liver concentrations of alphaTP affects its biliary secretion and the fatty acid compositions of hepatic and biliary lipids. Ten ovariectomized rats were assigned to two groups: five rats were implanted subcutaneously with time-release estradiol pellets (OXE; 25 microg/day/rat) and five with placebo (OXP). Twice daily rats were pair-fed a modified AIN-93G diet containing soybean oil. At 5 weeks, bile was collected via a bile cannula hourly for 8 hours during duodenal infusion of a lipid emulsion (565 micromol triolein and 396 micromol Na taurocholate/24 mL phosphate buffered saline, pH 6.45) at 3.0 mL/hr. During the 8 hour period, no difference was noted in the hourly rate of bile flow (0.95 mL/hr in OXE rats vs. 0.99 mL/hr in OXP rats). The biliary output of alphaTP for 8 hours was higher in OXE rats (51.6 +/- 3.6 nmol) than OXP rats (31.7 +/- 2.9 nmol). Likewise, the liver concentration of alphaTP was higher in OXE rats (81.9 +/- 3.5 nmol/g liver) than in OXP rats (53.3 +/- 7.4 nmol/g liver). The biliary secretion of phospholipids (PL) for 8 hours was significantly (P < 0.05) higher in OXE rats (55.1 +/- 4.9 micromol) than in OXP rats (42.3 +/- 4.7 micromol). Among the PL fatty acids, the outputs of 20:4 and 22:6n-3 were increased most markedly by estradiol replacement. The total outputs of 22:6n-3 for 8 hours in OXE and OXP rats were 2.95 +/- 0.20 micromol and 1.37 +/- 0.23 micromol, respectively. In the liver, the concentrations of PL 22:5n-3 and 22:6n-3 were elevated significantly in OXE rats. The present results suggest that estradiol may protect hepatic PL and membranes against oxidative damage by improving the liver status of alphaTP. PMID- 15539279 TI - Pectin and psyllium decrease the susceptibility of LDL to oxidation in guinea pigs. AB - These studies were undertaken to determine whether pectin (PE) and psyllium (PSY) intake affect the circulating levels of alpha-tocopherol and the susceptibility of low density lipoprotein (LDL) to oxidation. For that purpose, male Hartley guinea pigs were fed 19 g/100 g of a fat mix with a 2:1:1 ratio of saturated:polyunsaturated:monounsaturated fatty acids and 35 g/100 g total carbohydrate with 80% of the carbohydrate energy contributed by sucrose. Diets were identical in composition except for the fiber source: cellulose (control diet), PE, or PSY. Guinea pigs fed PE or PSY had 36% and 67% lower plasma cholesterol concentrations, respectively, compared with controls (P < 0.001). This plasma cholesterol lowering was associated with both very low density lipoproteins and LDL cholesterol fractions. Intake of PE or PSY resulted in 54% lower plasma triacylglycerol (TAG) concentrations compared with the control group (P < 0.001). LDL from PE and PSY fed guinea pigs contained fewer molecules of cholesteryl ester, and alpha-tocopherol concentrations in this particle were 49% and 66% higher, respectively, compared with controls. In addition, LDL from guinea pigs fed soluble fiber exhibited less susceptibility to oxidation than those from the control group, as determined by thiobarbituric acid-reactive substances formation. Hepatic free and esterified cholesterol were 32% lower and hepatic TAG was 25% lower in guinea pigs fed PE and PSY compared with controls. The data from these studies confirm that PE and PSY reverse the hyperlipidemia associated with high fat-sucrose diets and demonstrate a potential antioxidant effect of soluble fiber on circulating LDL. PMID- 15539280 TI - Biotin biochemistry and human requirements. AB - Human biotin turnover and requirements can be estimated on the basis of (1) concentrations of biotin and metabolites in body fluids, (2) activities of biotin dependent carboxylases, and (3) the urinary excretion of organic acids that are formed at increased rates if carboxylase activities are reduced. Recent studies suggest that the urinary excretions of biotin and its metabolite bisnorbiotin, activities of propionyl-CoA carboxylase and beta-methylcrotonyl-CoA carboxylase in lymphocytes, and urinary excretion of 3-hydroxyisovaleric acid are good indicators of marginal biotin deficiency. On the basis of studies using these indicators of biotin deficiency, an adequate intake of 30 microg (123 nmoles) of biotin per day is currently recommended for adults. The dietary biotin intake in Western populations has been estimated to be 35 to 70 microg/d (143-287 nmol/d). Recent studies suggest that humans absorb biotin nearly completely. Conditions that may increase biotin requirements in humans include pregnancy, lactation, and therapy with anticonvulsants or lipoic acid. PMID- 15539281 TI - Zinc-related metallothionein metabolism in bovine pulmonary artery endothelial cells. AB - Bovine pulmonary artery endothelial cells (BPAEC) were cultured in vitro under a variety of conditions to investigate how metallothionein (MT) might participate in zinc homeostasis. Experimental conditions included 10% serum to ensure that the in vitro environment would be a better reflection of the in vivo situation than with protein-free medium. MT was increased by acutely high zinc concentrations (100-200 micromol/L) in the extracellular environment. MT was relatively insensitive to moderate changes in zinc concentration (2-50 micromol/L), even after prolonged exposure for 7 to 12 days. BPAEC had reduced MT content when grown in medium containing serum that had been dialyzed to remove components with a molecular mass of less than 1,000, including zinc. Because the principal source of the major minerals in the experimental medium was not the serum, their concentrations in the final medium were not significantly influenced by serum dialysis. Restoring the zinc concentration in the medium containing the dialyzed serum did not restore MT content in BPAEC, suggesting that some small molecular weight molecule other than zinc established their basal MT content. This study did not identify these putative factors in serum, but hormones are likely candidates. Forty-eight-hour incubations of BPAEC with interleukin (IL-6) or dexamethasone increased cellular MT; however, 17beta-estradiol decreased MT, and IL-1 and adenosine 3',5'-cyclic phosphate (cAMP) had no discernible effect. We conclude that extracellular zinc concentrations have relatively little impact on the cellular concentrations of MT and zinc of BPAEC in vitro. Zinc homeostasis by BPAEC is not maintained by changing the MT concentration in response to changes in the extracellular zinc environment. (J. Nutr. Biochem. 10:00-00, 1999). PMID- 15539282 TI - "Antioxidant" (reducing) efficiency of ascorbate in plasma is not affected by concentration. AB - Ascorbate (vitamin C), an important dietary derived antioxidant, reportedly shows decreasing "antioxidant efficiency" with increasing concentrations in indirect radical trapping methods of antioxidant capacity. This study investigated the effect of concentration on antioxidant efficiency of ascorbate using a direct test of antioxidant capacity, the ferric reducing/antioxidant power test (FRAP assay). Results showed that the antioxidant efficiency factor of ascorbate was 2 and was constant over a wide concentration range in both plasma and pure aqueous solution. However, the absolute amount of ascorbate lost per unit of time increased with concentration. Furthermore, ascorbate was less stable in plasma than in aqueous solutions of similar pH and less stable in ethylenediamine tetraacetic acid (EDTA) than in heparinized plasma. Results indicate that previously reported concentration-dependent changes in antioxidant efficiency of ascorbate may have been caused by loss of ascorbate prior to and during testing, and by methodologic characteristics of indirect peroxyl radical trapping tests of antioxidant capacity. Therefore, it is suggested that the premise that the antioxidant efficiency of ascorbate is concentration-dependent is largely methodologically derived and does not reflect the antioxidant behavior of ascorbate per se. PMID- 15539283 TI - Omega-3 fatty acids alter lipoprotein subfraction distributions and the in vitro conversion of very low density lipoproteins to low density lipoproteins. AB - The purpose of this study was to determine the effects of a fish oil concentrate (FOC) on the in vitro conversion of very low density lipoproteins (VLDL) to intermediate (IDL) and low density lipoproteins (LDL). Six hypertriglyceridemic patients were randomly allocated to receive either placebo (olive oil) or FOC (1 g/14 kg body weight/day) for 4 weeks in a crossover study with a 4-week washout period. The FOC provided 3 g of eicosapentaenoic + docosahexaenoic acid per 70 kg of body weight, and it lowered plasma triglyceride and VLDL cholesterol levels by 35% and 42%, respectively. Decreases in the largest particles (VLDL(1)) were primarily responsible, with no effect noted in smaller VLDL particles (VLDL(2) and VLDL(3)). The FOC increased LDL cholesterol levels by 25% (P < 0.06) but did not affect LDL particle size. VLDL(1) and VLDL(3) were incubated in vitro with human postheparin lipases. Although triglycerides from both types of VLDL were hydrolyzed to the same extent with both treatments, particles isolated during the FOC phase were more readily converted into IDL and LDL than were control particles. These data suggest that the marine omega3 fatty acids may enhance the propensity of VLDL to be converted to LDL, partly explaining the decreased VLDL and increased LDL levels in FOC-treated patients. PMID- 15539284 TI - Olive oil hydroxytyrosol protects human erythrocytes against oxidative damages. AB - Hydroxytyrosol, the major representative phenolic compound of virgin olive oil, is a dietary component. Its possible protective effect on hydrogen peroxide (H(2)O(2))-induced oxidative alterations was investigated in human erythrocytes. Cells were pretreated with micromolar hydroxytyrosol concentrations and then exposed to H(2)O(2) over different time intervals. Subsequently, erythrocytes were analyzed for oxidative hemolysis and lipid peroxidation. Our data demonstrate that hydroxytyrosol prevents both oxidative alterations, therefore, providing protection against peroxide-induced cytotoxicity in erythrocytes. The effect of oxidative stress on erythrocyte membrane transport systems, as well as the protective role of hydroxytyrosol, also were investigated in conditions of nonhemolytic mild H(2)O(2) treatment. Under these experimental conditions, a marked decrease in the energy-dependent methionine and leucine transport is observable; this alteration is quantitatively prevented by hydroxytyrosol pretreatment. On the other hand, the energy-independent glucose transport is not affected by the oxidative treatment. The reported data give new experimental support to the hypothesis of a protective role played by nonvitamin antioxidant components of virgin olive oil on oxidative stress in human systems. PMID- 15539285 TI - Role of dietary lysine, methionine, and arginine in the regulation of hypercholesterolemia in rabbits. AB - These experiments were conducted to see whether the hypercholesterolemia produced by a diet enriched in lysine (Lys) and methionine (Met) can be reproduced by feeding these amino acids separately, and whether dietary arginine (Arg) counteracts their hypercholesterolemic effects. Another aim was to investigate the mechanisms involved in modulations of serum cholesterol levels by these amino acids. The results of this study, which were in agreement with the results of earlier experiments in our laboratory, showed that feeding a low-fat, cholesterol free, semipurified amino acid diet enriched with Lys + Met to rabbits caused a marked increase in serum total and low density lipoprotein cholesterol and apolipoprotein B levels, whereas a similar diet enriched in essential ketogenic amino acids (EketoAA) resulted in a more moderate increase in these parameters. Supplementing the diet with either Lys or Met alone was also less effective in inducing hypercholesterolemia than increasing levels of both amino acids. Dietary Arg partially counteracted the hypercholesterolemic effect of Lys + Met but not that of the EketoAA or of Lys and Met fed separately. The growth performance of rabbits fed the Lys + Met diet was inferior to that of those fed the other diets. Liver total phospholipid levels and the ratio of phosphatidylcholine to phosphatidylethanolamine were higher in rabbits fed the Lys + Met-enriched diet than in those animals fed a diet in which Arg was supplemented. In conclusion, our results indicate that high levels of both Lys and Met are needed to cause a maximum elevation of serum cholesterol and that the moderately antihypercholesterolemic effect of Arg is seen only when both amino acids are supplemented. They also suggest that these essential amino acids may affect cholesterol metabolism partly through alteration of liver phospholipids. PMID- 15539286 TI - Regulation of guinea pig hepatic acyl-coa:cholesterol acyltransferase activity by dietary fat saturation and cholesterol. AB - We measured the interactive effects of dietary cholesterol and fat on the regulation of hepatic acyl-CoA:cholesterol acyltransferase (ACAT) activity and its relationship to hepatic microsomal lipid composition in guinea pigs fed 15 g/100 g (w/w) fat diets (corn oil, olive oil, or lard) with 0.01, 0.08, 0.17, or 0.33 g/100 g (w/w) added cholesterol. Guinea pigs exhibited a dose dependent increase in hepatic microsomal ACAT activity, with increasing levels of cholesterol intake (P < 0.001) in all dietary fat groups. Animals fed monounsaturated olive oil had the highest hepatic ACAT activity with the exception of the 0.33 g/100 g cholesterol diet (P < 0.001). There were no differences in ACAT activity with intake of polyunsaturated corn oil or saturated lard. Dietary cholesterol resulted in increased microsomal free cholesterol (FC) concentrations in a dose dependent manner but had no effects on microsomal phosphatidylcholine (PC) concentrations. Guinea pigs fed olive oil generally had the highest microsomal FC/PC molar ratios, and hepatic ACAT activities correlated significantly with this parameter. After modification of the lipid compositions of the microsomes from guinea pigs fed the 12 test diets with FC/PC liposome treatment, microsomal ACAT activities remained significantly related to the microsomal FC/PC molar ratios, and dietary fat type did not affect this correlation. Our findings do not support the hypothesis that the stimulation of hepatic ACAT activity with cholesterol intake is enhanced by polyunsaturated fat intake. The data demonstrate that although dietary fat type and cholesterol amount have differential effects on hepatic ACAT activity, substrate availability, expressed as microsomal FC/PC molar ratio, is a major regulator of hepatic microsomal ACAT activity. PMID- 15539287 TI - Visible light-induced photooxidation of glucose sensitized by riboflavin. AB - We conducted this study to evaluate the oxidation of glucose induced by visible light in the presence of sensitizers such as methylene blue and flavins (i.e., flavin mononucleotide and riboflavin). The concentration of the sensitizers was similar to that of flavin in parenteral nutrients. The photooxidation of glucose sensitized by flavin mononucleotide or riboflavin was greater than that which was observed in the presence of methylene blue, whereas the isotopic effect of deuterium oxide (D(2)O) was enhanced more substantially in the presence of methylene blue than in the presence of flavins. These results show that methylene blue exerts its action through singlet oxygen and that at a high substrate concentration (as was used in this work) flavin mononucleotide and riboflavin act preferentially as type I sensitizers. In the flavin photosensitized processes, the presence of hydrogen peroxide, superoxide anion, and hydroxyl radical was demonstrated. The photooxidation of glucose is favored by an increase in pH, and it also depends on the energy absorbed by the system. By using a specific reagent for glucose (i.e., o-toluidine), it was possible to quantify the photoconversion of glucose. The results obtained in this work should be considered in the management of glucose-containing parenteral nutrients that are exposed to visible light in the presence of a multivitamin complex containing flavin mononucleotide. PMID- 15539288 TI - The function of cholesterol in embryogenesis. AB - Cholesterol is critical in embryonic development. Inhibition of cholesterol synthesis in experimental animals has caused a birth defect called holoprosencephaly (HPE), which is evidenced by cyclopia (one eye in the middle of the face), monorhinia (protruding single nose above the eye), absence of the pituitary gland, and central nervous system (CNS) abnormalities. In humans, an inherited defect in the cholesterol-synthesizing enzyme 7-dehydrocholesterol reductase depletes cholesterol and results in human HPE, termed Smith-Lemli-Opitz syndrome. In its most severe form, the syndrome leads to cyclopia, monorhinia, and lack of separation of cerebral hemispheres. The cause of the syndrome is a defect in a protein coded by the gene Sonic hedgehog (SHH). The protein SHH is expressed in the notochord of the CNS in the early embryo and is activated by being cleaved autocatalytically, with simultaneous covalent attachment of cholesterol to the N-terminal fragment, which is secreted by cells of the mesoderm layer, signaling the establishment of the neural midline cells. Thus, cholesterol is essential for proper signaling in the development of the normal embryo. PMID- 15539289 TI - Flavonoid transport by mammalian endothelial cells. AB - Despite the ever-growing body of literature reporting the effects of flavonoids on animals at both the cellular and systemic levels, one of the most basic questions-"Are the effects of flavonoids on animal cells initiated through their interaction with extracellular targets or intracellular targets?"-has yet to be addressed. Because many effects of flavonoids on cells can be detected within minutes of flavonoid application and because flavonoids diffuse across lipid membranes slowly or not at all, intracellular mechanisms would necessitate a flavonoid transport system for rapid flavonoid uptake. The specific aims of this investigation were (1) to determine if endothelial cells contain a mechanism that mediates rapid flavonoid uptake and (2) to provide evidence for or against the hypothesis that rapid flavonoid effects on endothelial cell synthesis of prostacyclin and endothelin are initiated through the interaction of flavonoids with intracellular targets. Data show that bovine and human aortic endothelial cells possess a transport system that mediates rapid uptake of the flavonoid morin and suggest that the flavonoid uptake system utilizes a variety of oxygenated phenolic compounds as substrates. Further investigation into flavonoid transport should expedite future investigation into the mechanisms of flavonoid actions, because it may allow research to focus on the cellular locations where flavonoids are concentrated. Although endothelial cells contain a mechanism for the rapid uptake of morin, data reported herein suggest that morin initiates its rapid effects on endothelial cell synthesis of prostacyclin and endothelin through an interaction with extracellular targets. PMID- 15539290 TI - Differential effects of dietary fat on chick plasma and liver composition and HMG CoA reductase activity. AB - The comparative effects of diet supplementation with 10% saturated fat rich in 12:0 and 14:0 fatty acids (coconut oil), without and with 1% added cholesterol, and with 10% unsaturated fat rich in n-3 polyunsaturated fatty acids (menhaden oil) on cholesterol metabolism in neonatal chicks were examined to clarify the different mechanisms of their hyper- and hypolipidemic action. Supplementation of coconut oil produced a significant hypercholesterolemia after 7 days of treatment, with a similar increase in the amount of both free and esterified cholesterol. Supplementation of coconut oil plus cholesterol produced a higher increase of plasma cholesterol levels (approximately two to three times higher than those found with standard diet). However, supplementation of menhaden oil induced a significant decrease in total cholesterol after only 2 weeks of treatment. Levels of plasma triglycerides did not change by coconut oil addition to the diet, but a significant increase was observed after coconut oil plus cholesterol feeding. Menhaden oil produced a transient decrease in plasma triglycerides. Hepatic 3-hydroxy-3-methylglutaryl-CoA reductase activity did not change with coconut oil treatment. However, both coconut oil plus cholesterol and menhaden oil supplemented diets drastically decreased reductase activity after 1 week of dietary manipulation. These results show that different nutrients with the same inhibitory effect on reductase activity produced opposite effects on plasma cholesterol content, suggesting the existence of important differences in the regulatory mechanisms implied in cholesterol biosynthesis and its accumulation in plasma. PMID- 15539291 TI - A method for the determination of glucose synthesis in isolated bovine hepatocytes. AB - A simple method for determining glucose synthesis from radiolabeled precursors in isolated bovine hepatocytes using ion exchange resins is presented. This method allows processing of multiple small volume samples using suspensions of anion and cation exchange resins rather than traditional stacked column separation methods. Hepatocytes were isolated from calf liver by collagenase perfusion of the caudate lobe and were incubated with (14)C-labeled lactate or propionate as gluconeogenic substrates. Glucose synthesis was determined in an aliquot of cell suspension that was vortexed with a slurry of anion exchange (acetate form) resin, followed by a slurry of cation exchange resin. Newly synthesized, labeled glucose was recovered in the supernatant after centrifugation and quantitated by scintillation counting. Using this procedure, more than 98% of the unused labeled precursor was bound to the ion exchange resin and essentially 100% of a labeled glucose tracer was recovered in the supernatant. Pretreatment of hepatocyte suspensions with glucose oxidase was shown to eliminate the accumulation of radioactivity in the supernatant, thus confirming the specificity of this technique for measurement of newly synthesized glucose. This method was sensitive to changes in the rate of hepatic gluconeogenesis that resulted from changes in substrate concentration or the addition of glucagon or fatty acids to the hepatocyte incubations. PMID- 15539292 TI - Early and advanced glycation end-products are increased in dietary copper deficiency. AB - The hypothesis that nonenzymatic glycosylation of proteins (glycation) contributes to damage associated with dietary copper deficiency has depended largely on indirect evidence. Thus far, the observation of an elevated percentage of glycated hemoglobin in copper-deficient rats has provided the only direct evidence of an increase in glycation. We sought further direct evidence of increased glycation in copper deficiency. Male weanling rats were fed a copper adequate (CuA, 6.4 mg Cu/kg diet) or copper-deficient diet (CuD, 0.4 mg Cu/kg diet) for 5 weeks. Rats fed the CuD diet were copper deficient as judged by depressed organ copper concentrations and a variety of indirect indices. Measurements of hemoglobin A(1) and serum fructosamine (both early glycation end products) as well as serum pentosidine (an advanced glycation end-product) indicated that all three compounds were elevated in CuD rats relative to CuA rats. This finding further supports the view that glycation is enhanced and thus may contribute to defects associated with dietary copper deficiency. PMID- 15539293 TI - Mechanisms of absorption of caseinophosphopeptide bound iron. AB - Binding iron (Fe) to the 1-25 caseinophosphopeptide obtained from enzyme hydrolysis of beta casein (beta CPP) improves Fe bioavailability in the rat. To assess the mechanisms involved in its absorption, a perfused, vascularized duodenal rat loop model was used in controls and in Fe-deficient (bleeding of 25% blood volume) rats. Inhibitors of oxidative phosphorylation [2-4 dinitrophenol (DNP)] and/or of endocytosis [phenylarsine oxide (PAO)] were added to the perfusion solution containing 50 microM Fe as beta CPP bound Fe (Fe-beta CPP) or gluconate (Fe Gluc). Fe-beta CPP enhanced Fe uptake, reduced mucosal storage, and improved net absorption both in controls and in deficient animals. DNP reduced uptake, mucosal storage, and net absorption by the same percentage in Fe-beta CPP and Fe Gluc perfused rats in both control and Fe-deficient animals. PAO decreased uptake, mucosal storage, and net absorption of Fe-beta CPP but not of Fe Gluc. At the end of the experiment Fe serum levels were increased only in Fe Gluc animals. These results confirm the improved bioavailability of beta CPP bound Fe. They suggest that at least part of its absorption can occur by a different pathway than usual Fe salts. Fe-beta CPP can be taken up by endocytosis and absorbed bound to amino acids or peptides. PMID- 15539294 TI - Formation of antioxidants from (-)-epigallocatechin gallate in mild alkaline fluids, such as authentic intestinal juice and mouse plasma. AB - The oxidative dimerization of (-)-epigallocatechin gallate (EGCG), the major catechin of tea leaves (Camellia sinensis L.), in authentic intestinal juice (pH 8.5) and mouse plasma (pH 7.8) was investigated. EGCG was unstable in the alkaline solutions over pH 7.4. The content of EGCG was decreased to 19.4% and 60.7% at 5 minutes in the intestinal juice and plasma, respectively. Three products-P-1 (theasinensin A), P-2 (a new dimerized product reported in a previous paper), and P-3 (theasinensin D, a rotational isomer of P-1)-were detected in these fluids. The sum of the molar contents of the three products formed after 5 minutes of incubation at 37 degrees C corresponded to 35.1% and 21.9% of the degraded molar content of EGCG, respectively. These dimerization products of EGCG would be formed by the dehydrogenation and decarboxylation of EGCG under oxidative conditions in alkaline solutions. The formation of P-2 was greater than that of P-1 and of P-3 at 30 minutes of incubation in the intestinal juice and mouse plasma. Fe(2+)-chelating activities of the three products were much higher than that of EGCG, and superoxide anion radical-scavenging activity of P-2 was also significantly higher than that of EGCG. The absorbance of P-2 administered to male ddY mice was studied. The content of P-2 in mouse plasma was less than that of administration of EGCG, but P-2 was absorbed quickly within 30 minutes and metabolized slowly. These dimerization products of EGCG are expected to contribute to in vivo antioxidative activities enhanced by tea drinking. PMID- 15539295 TI - Diet deficient in alpha-linolenic acid alters fatty acid composition and enzymatic properties of Na+, K+-ATPase isoenzymes of brain membranes in the adult rat. AB - The effects of dietary (n-6)/(n-3) polyunsaturated fatty acid balance on fatty acid composition, ouabain inhibition, and Na(+) dependence of Na(+), K(+)-ATPase isoenzymes of whole brain membranes were studied in 60-day-old rats fed over two generations a diet either devoid of alpha-linolenic acid [18:3(n-3)] (sunflower oil diet) or rich in 18:3(n-3) (soybean oil diet). In the brain membranes, the sunflower oil diet led to a dramatic decrease in docosahexaenoic acid [22:6(n-3)] membrane content. The activities of Na(+), K(+)-ATPase isoenzymes were discriminated on the basis of their differential affinities for ouabain and their sensitivity to sodium concentration. The ouabain titration curve of Na(+), K(+) ATPase activity displayed three inhibitory processes with markedly different affinity [i.e., low (alpha1), high (alpha2), and very high (alpha3)] for brain membranes of rats fed the sunflower oil diet, whereas the brain membranes of rats fed the soybean oil diet exhibited only two inhibitory processes, low (alpha1) and high (alpha2' = alpha2 + alpha3). Regardless of the diet, on the basis of the Na(+) dependence of Na(+), K(+)-ATPase activity, three isoenzymes were found: alpha1 form displaying an affinity 1.5- to 2-fold higher that of than alpha2 and 3-fold higher that of alpha3. In rats fed the sunflower oil diet, alpha2 isoenzyme exhibited higher affinity for sodium (Ka = 8.8 mmol/L) than that of rats fed the soybean oil diet (Ka = 11.7 mmol/L). These results suggest that the membrane lipid environment modulates the functional properties of Na(+), K(+) ATPase isoenzymes of high ouabain affinity (alpha2). PMID- 15539296 TI - Biotin enhances glucose-stimulated insulin secretion in the isolated perfused pancreas of the rat. AB - The effects of biotin on insulin secretion in pair-fed control rats and biotin deficient rats were investigated using the method of isolated pancreas perfusion. Isolated pancreas perfusion was performed using 20 mM glucose, 10 mM arginine, and 20 mM glucose plus various concentrations of biotin (20 mM glucose + biotin solution) as stimulants of insulin secretion. The insulin response to 20 mM glucose in biotin-deficient rats was approximately 22% of that seen in control rats. The level of the insulin response to 10 mM arginine was also significantly lower in biotin-deficient rats than in control rats. These results indicate that insulin release from the pancreas was disturbed in biotin-deficient rats. The insulin responses to 20 mM glucose + 1 mM biotin in biotin-deficient and control rats increased to 165% and 185%, respectively, of that to 20 mM glucose. These biotin-induced increases in glucose-stimulated insulin release were evident within the first few minutes of the infusion. An enhancement of the arginine induced insulin response in control rats was not found when arginine and biotin was administered. These results suggest that biotin may play an important role in the mechanism by which glucose stimulates insulin secretion from the beta cells of the pancreatic islets. PMID- 15539297 TI - Mechanism for the increased skeletal muscle protein degradation in the obese Zucker rat. AB - Obese (fa/fa) Zucker rats showed a decreased protein content in skeletal muscle compared with their lean counterparts. This was associated with both a decrease in the fractional rate of protein synthesis and an enhanced fractional rate of protein degradation in skeletal muscle, as studied by pre-loading body proteins with (14)C-bicarbonate. The increased protein degradation could be the result of the clearly enhanced expression for several transcripts of the ubiquitin genes in skeletal muscle. The results suggest that the increased protein degradation in the skeletal muscle of the obese animals may be due to increased activity of the ubiquitin-dependent proteolytic system. PMID- 15539298 TI - Antiarrhythmic fatty acids and antioxidants in animal and cell studies. AB - From the animal and cellular studies that will be discussed in this review, it is apparent that dietary fatty acids and antioxidants play an important role in influencing the development of ventricular tachycardia and potentially lethal ventricular fibrillation. It is this latter disturbance to the rhythmic beating of the heart that is responsible for much of the mortality from coronary heart disease. It is now recognized that diets high in certain polyunsaturated fatty acids (PUFAs) and diets containing antioxidants can afford considerable protection to the heart with regard to the generation of disorders of contractile rhythmicity. The mechanism by which such dietary components confer their cardioprotective effects are now being intensively investigated, particularly with respect to their possible effects on the molecular mechanisms underlying the excitation-contraction coupling process of the myocardial cell. This overview will cover recent studies that have focused on the antiarrhythmic role of PUFAs, particularly those of the n-3 (or omega 3) class with emphasis on experiments performed using laboratory animals, isolated heart preparations, and isolated heart cells (cardiomyocytes). The role of free radicals (reactive oxygen species) and antioxidants in disorders of cardiac rhythm also will be addressed within the perspective of reperfusion injury to the myocardium following ischemia. Emphasis will be placed on the cardioprotective role of nutritional factors and components and the possible cellular mechanisms by which such components may act. PMID- 15539299 TI - Plasma protein carbonyl concentration is not enhanced by chronic intake of high protein diets in adult rats. AB - We tested the hypothesis of whether high dietary protein intake is linked to oxidative stress as measured by the concentration of reactive carbonyl residues in plasma proteins. Three groups of male Wistar rats ( approximately 230 g, n = 10) were fed either 15% (15C), 30% (30C), or 60% (60C) casein diets over a period of 18 weeks. For comparison, a vitamin E deficient diet (60C-E) based on diet 60C was given to an additional group to provoke oxidative stress. Concentrations of alpha-tocopherol in plasma and of reactive carbonyl residues in total plasma proteins were measured by high performance liquid chromatography using fluorescence and by diode array detection after 2,4-dinitrophenylhydrazine reaction, respectively. After 1 week the concentration of reactive carbonyl residues in plasma proteins was found to be significantly (P < 0.05) higher in the 60C and 60C-E groups ( approximately 2.7 nmol/mg protein) compared with the 15C and 30C groups ( approximately 1.7 nmol/mg protein). After 14 weeks the 15C (3.4 +/- 1.2 nmol/mg protein) and 60C-E groups (3.9 +/- 1.7 nmol/mg protein) showed a significantly increased concentration of reactive carbonyl residues in plasma protein compared with the 30C and 60C groups (2.5 +/- 1.0 nmol/mg protein; 2.6 +/- 0.8 nmol/mg protein). As expected, chronic vitamin E deficiency (60C-E) resulted in significantly decreased alpha-tocopherol concentrations (3.91 +/- 2.42 micromol/mL vs. 31.3 +/- 4.8 micromol/mL) and a higher concentration of reactive carbonyl residues in plasma proteins. These results do not support the hypothesis that a chronic intake of high-protein diets leads to oxidative stress in adult rats. However, in the non-adapted state (1 week) a high protein intake contributes to oxidative modifications of protein-bound amino acid residues. PMID- 15539300 TI - Feeding the nitric oxide synthase inhibitor L-N(omega)nitroarginine elevates serum very low density lipoprotein and hepatic triglyceride synthesis in rats. AB - This study was conducted to study the influence of dietary L N(omega)nitroarginine (L-NNA), a nitric oxide (NO) synthase inhibitor, on serum lipids and lipoproteins and on the activities of enzymes related to lipid metabolism in rats. Feeding rats a diet containing 0.2 g/kg L-NNA for 5 weeks elevated serum concentrations of triglyceride, cholesterol, phospholipid, and free fatty acid and reduced serum nitrate (an oxidation product of NO). The elevation in serum triglyceride was mainly due to the elevation in very low density lipoprotein (VLDL) triglyceride. Contents of cholesterol and phospholipid in the VLDL fraction also were elevated by L-NNA. L-NNA treatment caused significantly higher activity of hepatic microsomal phosphatidate phosphohydrolase (the rate-limiting enzyme in triglyceride synthesis) and lower activity of hepatic carnitine palmitoyltransferase (the rate-limiting enzyme in fatty acid oxidation). Activities of hepatic enzymes responsible for fatty acid synthesis such as glucose-6-phosphate dehydrogenase, malic enzyme, and fatty acid synthase were unaffected by L-NNA. The activity of hepatic microsomal phosphocholine cytidyltransferase (the rate-limiting enzyme in phosphatidylcholine synthesis) was reduced significantly by L-NNA. Our results suggest that lower NO production caused the elevations in hepatic triglyceride synthesis by higher esterification of fatty acid and lower fatty acid oxidation, leading to an enrichment of VLDL triglyceride. PMID- 15539301 TI - A possible role for cyclins in the zinc requirements during G1 and G2 phases of the cell cycle. AB - Zinc has been shown to be required for the passage of cells through the mid-G1 phase of the cell cycle and for differentiation of myoblasts. However, it has been suggested that zinc has other roles during the cell cycle. The experiments reported here indicate that readily available zinc is not required for DNA synthesis per se but is needed for a process contemporaneous with the S phase and required for subsequent progress of the cells through G2 and mitosis. The G1 and S/G2 requirements for zinc showed virtually identical sensitivities to zinc deprivation. Each of the above requirements for zinc coincides with the induction of specific cyclin mRNAs, and the concentrations of these mRNAs have now been shown to decrease in the absence of adequate zinc. This is the first study to indicate a possible common factor underlying the requirement for available zinc during both cell replication and differentiation. PMID- 15539302 TI - Effect of dietary fat on whole body fatty acid synthesis in weanling rats. AB - The effect of dietary fat on body composition, whole body lipogenesis, and enzyme activity was measured in rats over the first 16 weeks post-weaning. Rats were fed either a low fat (5% w/w fat) or high fat (20% w/w fat) diet for the first 4 weeks. After this time all rats were fed the low fat diet. The results showed no significant effect of diet on the rate of fat synthesis over the first 8 weeks of the experiment. However, the activities of the enzymes of fatty acid synthesis [glucose 6-phosphate dehydrogenase, malic enzyme, adenosine triphosphate-citrate lyase, acetyl-coenzyme A carboxylase (ACCX), fatty acid synthetase] were dependent on the age and dietary status of the animals. The exact pattern depended on the specific enzyme and the tissue source. No significant differences in pyruvate dehydrogenase (PDH) activity were observed. Mathematical analysis of the enzyme activities suggested that ACCX and PDH were the most likely sites of fat synthesis regulation. In addition, an examination of body composition and overall weight retention showed that the "weight increasing" effect of a high fat diet could be completely reversed by subsequent feeding of a low fat diet. However, the reversal required an additional 12 weeks. Interestingly, at this time the rats switched from a high fat to a low fat diet had a lower body weight and lower body fat content than rats fed a low fat diet throughout the course of the experiment. PMID- 15539303 TI - Characterization of brush border membrane-bound alkaline phosphatase activity in different segments of the porcine small intestine. AB - This study was conducted to characterize enterocyte apical membrane-bound alkaline phosphatase activity in different segments of the porcine small intestine. Duodenal, jejunal, and distal ileal segments were isolated from three 26-kg pigs and enterocyte brush border membrane, enriched between 19- and 24-fold in sucrase specific activity, was prepared by Mg(2+) precipitation and differential centrifugation. With P-nitrophenyl phosphate as substrate, the optimum pH for porcine brush border membrane-bound alkaline phosphatase activity was defined to be 10.5 for all three segments. At the optimal pH, the kinetics of membrane-bound alkaline phosphatase were determined for the three intestinal segments. The affinity of this enzyme (K(m), mM) in the jejunum (0.64 +/- 0.07) was four times greater than that in the duodenum (2.75 +/- 0.59) and the distal ileum (2.71 +/- 1.14). These results indicate that different isomers of membrane bound alkaline phosphatase might have been expressed in different segments of porcine small intestine. The maximal specific activity (V(max), micromol/mg protein . min) of this enzyme was highest in the duodenal (7.74 +/- 0.95), intermediate in the jejunal (4.31 +/- 0.18), and lowest in the distal ileal (3.53 +/- 0.84) brush border membrane. Therefore, the maximal specific activity of brush border membrane-bound alkaline phosphatase along the intestinal longitudinal axis in growing pigs decreases from the duodenum toward the distal ileum. PMID- 15539304 TI - Fatty acids alter time dependent loss of apolipoprotein E expression by primary cultures of rat hepatocytes. AB - Although the phenomenon of intracellular apolipoprotein E (apoE) degradation has been reported in other cell types, the fate of newly synthesized apoE in the liver is not well understood. In the present study, we examined the expression (the balance of synthesis, secretion, and degradation) of apoE in primary cultures of rat hepatocytes and compared it with albumin, a typical secretory protein. Synthesis and secretion of [(35)S]apoE was diminished in primary hepatocytes cultured for more than 2 days, in agreement with an observed decrease in apoE mRNA. Cells cultured for 1 day and labeled for up to 4 hours secreted total protein, apoE, and albumin, linearly. The apparent rates of synthesis for apoE and albumin were similar (1,158 vs. 1,334 dpm/mg/min) but rates of their secretion differed significantly (225 vs. 1,159 dpm/mg/min). Pulse-chase experiments indicated that cell-associated [(35)S]albumin was secreted without degradation, whereas significant quantities of newly synthesized apoE were degraded. The overall synthesis and secretion of total proteins, including secretion of apoE, was enhanced by oleic acid (1 mmol/L). However, this effect may not be limited to oleic acid because other fatty acids showed a similar effect on apoE mRNA abundance. In control cells, apoE was found to associate with high density lipoproteins predominantly, although the fraction associated with very low density lipoprotein was increased in hepatocytes incubated with oleic acid. Overall, the findings from this study suggest that the level of apoE expression by primary hepatocytes is dependent on the age of the culture. The study also indicates that the phenomenon of apoE degradation occurs in primary hepatocytes. PMID- 15539305 TI - Glutamine metabolism by lymphocytes, macrophages, and neutrophils: its importance in health and disease. AB - Many aspects of the cell biology of lymphocytes, macrophages, and neutrophils have been studied extensively. Our recent work on these cells has investigated how fuel metabolism, especially glutamine metabolism, is related to the specific function of these cells in the inflammatory response. The high rate of glutamine utilization and its metabolism in such immune cells has raised the question of why glutamine is responsible for these functions. The macrophage has access to a variety of metabolic fuels both in vivo and in vitro. The quantitatively important role of glutamine in the processes of free radical and cytokine production has been established in our laboratories. Our current understanding of the rate of utilization and the pathway of metabolism of glutamine by cells of the immune system raises some intriguing questions concerning therapeutic manipulation of utilization of this amino acid, specifically the phagocytic and secretory capacities of cells of the defense system can be beneficially altered. PMID- 15539306 TI - Coconut oil induces short-term changes in lipid composition and enzyme activity of chick hepatic mitochondria. AB - We studied the short-term effects of a 20% coconut oil supplementation to the chick diet on lipid composition of liver and hepatic mitochondria, and changes that occurred in mitochondrial-associated enzymes as a result of this diet. No significant differences were observed in the lipid contents of liver when young chicks were fed the experimental diet, whereas hepatic mitochondria rapidly changed in response to this diet. Total cholesterol significantly increased in mitochondria at 24 hours of coconut oil diet feeding and decreased when dietary treatment was prolonged for 5 to 14 days. Changes in total mitochondrial phospholipids showed an inverse profile. A significant decrease in phosphatidylethanolamine and an increase in sphingomyelin were found at 24 hours. The cholesterol/phospholipid molar ratio significantly and rapidly (24 hours) increased in mitochondria from treated animals. Cytochrome oxidase activity drastically increased after 24 hours of experimental diet feeding and lowered to the control values when dietary manipulation was prolonged for 5 to 14 days. ATPase activity showed an inverse profile. Changes in cytochrome oxidase activity were parallel to changes in the cholesterol/phospholipid molar ratio, whereas changes in ATPase activity showed an inverse correlation with changes in this molar ratio. To our knowledge, this is one of the first reports on the very rapid response (24 hours) of mitochondrial lipid composition and function to saturated fat feeding. PMID- 15539307 TI - Can a glutamate-enriched diet counteract glutamine depletion in endotoxemic rats? AB - The study evaluated whether a glutamate-enriched diet would restore glutamine tissue pools and maintain tissue trophicity in endotoxemic rats. For this purpose, young male Sprague-Dawley rats received an intraperitoneal injection of lipopolysaccharide (LPS) from Escherichia coli at 3 mg/kg body weight. After 24 hours of food deprivation, the rats were enterally refed for 48 hours using Osmolite enriched with glutamate at 4 g/kg/d (LPS-Glu group, n = 7) or glycine isonitrogenous to glutamate (LPS-Gly group, n = 7). A control group (healthy group, n = 7) had free access to a standard rodent diet. Tissue weights and protein contents were significantly lower in both LPS-treated groups than in the healthy group. No plasma or tissue accumulation of glutamate was observed except in the liver. Glutamine concentrations were increased in the jejunum, liver, and plasma in the LPS-Glu group versus the other two groups (P < 0.05). Conversely, they were depleted in muscles of the endotoxemic groups versus the healthy group (P < 0.05). Villus height was significantly greater in the LPS-Glu group than in the LPS-Gly group in the jejunum (P < 0.05), but not in the ileum. In conclusion, a glutamate-enriched diet administered enterally to endotoxemic rats can counteract glutamine depletion in the splanchnic area but not in muscles. In addition, glutamate displayed a trophic effect restricted to the jejunum. PMID- 15539308 TI - Effect of saturated, omega-3 and omega-6 polyunsaturated fatty acids on myocardial infarction. AB - Dietary fatty acids have cholesterol lowering, antiatherogenic, and antiarrhythmic properties that decrease the risk of myocardial infarction (MI). This study was designed to study the effects of various oils rich in either polyunsaturated (omega-3 or omega-6) fatty acids (PUFA) or saturated fatty acids (SFA) on the severity of experimentally induced MI. Male albino Sprague-Dawley rats (100-150 g; n = 20) were fed diets enriched with fish oil (omega-3 PUFA), peanut oil (omega-6 PUFA), or coconut oil (SFA) for 60 days. Experimental MI was induced with isoproterenol. Mortality rates; serum enzymes aspartate amino transferase; alanine amino transferase; creatine phosphokinase (CPK); lipid profiles in serum, myocardium, and aorta; peroxide levels in heart and aorta; activities of catalase and superoxide dismutase; and levels of glutathione were measured. The results demonstrated that mortality rate, CPK levels, myocardial lipid peroxides, and glutathione levels were decreased in the omega-3 PUFA treated group. Maximum increase in parameters indicative of myocardial damage was seen in the coconut oil group. These findings suggest that dietary omega-3 PUFA offers maximum protection in experimentally induced MI in comparison to omega-6 PUFA and SFA enriched diets. SFA was found to have the least protective effect. PMID- 15539309 TI - Uptake of vitamin A in macrophages from physiologic transport proteins: role of retinol-binding protein and chylomicron remnants. AB - Vitamin A plays an important role in reducing infectious disease morbidity and mortality by enhancing immunity, an effect that is partly mediated by macrophages. Thus, knowing how these cells take up vitamin A is important. The results in the present study demonstrate that J774 macrophages efficiently take up chylomicron remnant retinyl esters and retinol-binding protein (retinol-RBP) bound retinol by specific and saturable mechanisms. The binding of (125)I-RBP to plasma membrane vesicles demonstrated that the macrophage receptor had a similar binding affinity, as was discovered previously for other cells. The B(max) for the macrophages was smaller than the values reported for placenta, bone marrow, and kidney, but larger than that reported for liver. The J774 cells also bound and took up [(3)H]retinol-RBP. Approximately 50 to 60% of the uptake may compete with excess unlabeled retinol-RBP and approximately 30 to 40% with excess transtyrethin. Following the uptake of [(3)H]retinol-RBP, an extensive esterification occurred: After 5 hours of incubation, 77.8 +/- 3.9% (SD; n = 3) of the cellular radioactivity was recovered as retinyl esters. The J774 cells also demonstrated saturable binding of chylomicron remnant [(3)H]retinyl esters, and a continuous uptake at 37 degrees C followed by an extensive hydrolysis of the retinyl esters. Binding could be inhibited by approximately 50% by excess unlabeled low density lipoprotein (LDL). In addition, lipoprotein lipase increased the binding of chylomicron remnant [(3)H]retinyl esters by approximately 30% and the uptake of chylomicron remnant [(3)H]retinyl ester by more than 300%. Furthermore, because sodium chlorate reduced binding with 40% and uptake with 55%, the results suggest that proteoglycans are involved in the uptake. Thus, the results suggest that both LDL receptor and LDL-related protein are involved in the uptake of chylomicron remnant [(3)H]retinyl ester in macrophages. PMID- 15539310 TI - Nutritional value of proteins from edible seaweed Palmaria palmata (dulse). AB - Palmaria palmata (Dulse) is a red seaweed that may be a potential protein source in the human diet. Its protein content, amino acid composition, and protein digestibility were studied with algae collected every month over a 1-year period. Significant variations in protein content were observed according to the season: The highest protein content (21.9 +/- 3.5%) was found in the winter-spring period and the lowest (11.9 +/- 2.0%) in the summer-early autumn period. Most of the essential amino acids were present throughout the year. After 6-hour in vitro digestion in a cell dialysis using porcine pepsin and porcine pancreatin, the digestibility of proteins from Palmaria palmata crude powder, represented by dialyzed nitrogen, was estimated at 29.52 +/- 1.47%. Relative digestibility was 56%, using casein hydrolysis as 100% reference digestibility. In vitro digestibility of proteins extracted in water was analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis using either bovine trypsin, bovine chymotrypsin, pronase from Streptomyces griseus, or human intestinal juice. Dulse proteins were hydrolyzed to a limited extent, which confirmed a rather low digestibility. Hydrolysis rate was higher with trypsin and lower with chymotrypsin compared with the two other enzymatic systems, pronase and intestinal juice, respectively. The association of algal powder and protein extract to casein and bovine serum albumin, respectively, produced a significant decrease in the hydrolysis rate of the standard proteins. In conclusion, the digestibility of Palmaria palmata proteins seems to be limited by the algae non proteic fraction. PMID- 15539311 TI - Interrelationships between low density lipoprotein receptor defect, serum fatty acid composition, and serum cholesterol concentration. AB - It is known that, in the general human population, serum fatty acid composition is correlated with serum triacylglycerol and cholesterol concentrations. The goal of the present study was to analyze whether the same is true of individuals who have a low density lipoprotein receptor (LDL-R) defect. Concentrations of 16 different fatty acids, cholesterol, triacylglycerol, and major lipoproteins in serum were determined in eight individuals who had (FH-North Karelia), the most common LDL-R defect in Finland, which causes familial hypercholesterolemia, and in their 30 relatives belonging to a single large pedigree as controls. The average number of double bonds (i.e., degree of desaturation) in serum fatty acids correlated negatively with the concentrations of serum total cholesterol (r = 0.27, P < 0.05) and total triacylglycerol (r = -0.71, P < 0.001) and positively with the number of fish meals per week (r = 0.50, P < 0.01), which was analyzed in all pedigree members jointly. These effects were similar in individuals having LDL-R defect, in which group the correlation coefficients were -0.31 (P = NS), 0.99 (P < 0.001), and 0.79 (P = NS) for serum total cholesterol, triacylglycerol, and weekly fish meals, respectively. Thus, LDL-R defect does not impair the correlation between serum fatty acid composition and serum triacylglycerol concentration. This result is in agreement with dietary studies that have shown that familial hypercholesterolemia patients respond very favorably to dietary therapy. PMID- 15539312 TI - Comparative content of total polyphenols and dietary fiber in tropical fruits and persimmon. AB - Recent studies have shown that dietary fiber and polyphenols of vegetables and fruits improve lipid metabolism and prevent the oxidation of low density lipoprotein cholesterol (LDL-C), which hinder the development of atherosclerosis. The goal of this study was to measure the total polyphenol and dietary fiber contents of some tropical fruits (i.e., pineapple, wax apple, rambutan, lichi, guava, and mango) and compare the results to the content of these substances in the better characterized persimmon. It was found that lichi, guava, and ripe mango (cv. Keaw) have 3.35, 4.95, and 6.25 mg of total polyphenols in 100 g fresh fruit, respectively. This is significantly higher than in persimmon, pineapple, wax apple, mature green mango, and rambutan [P < 0.0005 for pineapple (Smooth Cayene variant), wax apple, persimmon, rambutan, mature green mango (cv. Keaw); the value of P < 0.001 is found only for pineapple (Phuket, Queen variant)]. The same relationship was observed for the contents of gallic acid and of dietary fiber. It can be supposed that among the studied fruit, lichi, guava, and ripe mango may be preferable for dietary prevention of atherosclerosis. PMID- 15539313 TI - Antioxidant effects of fructosyl arginine, a Maillard reaction product in aged garlic extract. AB - The amino-carbonyl (Maillard) reaction of amino acids with sugars is a nonenzymatic browning reaction that takes place during the processing, cooking, and storage of foods. Maillard reaction products (MRPs) have been shown to possess interesting chemical and biological properties including antimutagenic and antioxidant activity. In this study, we determined the antioxidant effects of fructosyl arginine (Fru-Arg), a MRP in aged garlic extract. Low density lipoprotein (LDL) was incubated with Cu(2+) at 37 degrees C and 5% CO(2) for 24 hours, which resulted in an increase of thiobarbituric acid reactive substances (TBARS) indicating lipid peroxidation. Coincubation of Cu(2+) with Fru-Arg and LDL resulted in a significant inhibition of TBARS formation. Pulmonary artery endothelial cells (PAEC) were exposed to 0.1 mg/mL oxidized LDL (Ox-LDL) at 37 degrees C and 5% CO(2) for 24 hours. Lactate dehydrogenase (LDH) release, as an index of cell membrane damage, and TBARS were measured. Ox-LDL caused an increase of LDH release and TBARS formation. Pretreatment of PAEC with Fru-Arg inhibited these changes. Murine macrophages were incubated with Ox-LDL, and the release of peroxides was measured using a fluorometric assay. Ox-LDL caused an increased release of peroxides. Coincubation of macrophages with Fru-Arg and Ox-LDL inhibited the release of peroxides dose-dependently. In a cell free system, Fru Arg was shown to scavenge hydrogen peroxide. These data suggest that Fru-Arg is a potent antioxidant, and thus may be useful for the prevention of atherosclerosis and other disorders associated with oxidative stress. PMID- 15539314 TI - Nutritional support for burn injuries. AB - There are approximately 2,000,000 burns each year in the United States that require medical treatment. Fortunately, most of these can be treated on an outpatient basis; however, 100,000 patients are hospitalized yearly. The mortality rate is higher in patients with large burns or smoke inhalation injury or both, or in patients otherwise compromised by age or concomitant disease. Thermal injury leads to suppression of the immune response, and infectious morbidity is common in all burn patients. In addition, the injury and recovery processes are attended by tremendous catabolism of host tissues, leaving patients debilitated and functionally impaired. Medical nutrition therapy plays a key role in the support of the burn patient, supporting the immune system and blunting the hypermetabolic response. PMID- 15539315 TI - Induction of tissue transglutaminase expression by propionate and n-butyrate in colon cancer cell lines. AB - Short-chain fatty acids (SCFAs) have been demonstrated to induce differentiation and/or apoptosis in colon cancer cells. A close correlation between tissue transglutaminase (tTG) expression and differentiation and/or apoptosis has been suggested in many cell lineages. However, the effects of SCFAs on tTG expression in colon cancer cells have not yet been reported. In this report, the relationship between cytosolic tTG levels and differentiation state was investigated in six human colon cancer cell lines. Effects of four kinds of SCFAs (acetate, propionate, n-butyrate, and isobutyrate) on the expression of tTG then were investigated in association with their effects on apoptosis induction. High expression of tTG protein and mRNA were found in SW480 and WiDr cell lines, which exhibited well differentiated phenotypes. tTG expression was hardly detectable in the less differentiated cell lines COLO201, COLO320DM, and CW-2. However, n butyrate and propionate significantly increased cytosolic tTG levels at concentrations above 0.5 mM in these less differentiated colon cancer cells. n Butyrate and propionate induced growth suppression and apoptosis in these cell lines at concentrations that can induce tTG expression. Acetate and isobutyrate did not induce tTG expression or growth suppression at concentrations up to 8 mM. In conclusion, tTG induction by propionate and n-butyrate was suggested to be closely linked to their differentiation- and apoptosis-inducing effects in colon cancer cells. These findings may explain the mechanisms by which dietary fiber show preventive effects against colon carcinogenesis. PMID- 15539316 TI - Effects of a high soy protein diet on intestinal polyamines and ornithine decarboxylase activity in rats. AB - This study was performed to determine whether intestinal luminal polyamine concentrations are affected by a high soy protein diet when compared with a high casein diet or a normoprotein casein diet. We also determined the effects of these diets, with differences in polyamines content, on mucosal polyamines and ornithine decarboxylase (ODC) activity to assess cell proliferation. Three groups of eight male Wistar rats were fed either a 50% soy protein diet, a 50% casein diet, or an 18% casein diet as a control. After 4 weeks of feeding, both intestinal content and mucosa were recovered. Polyamines were assayed by high performance liquid chromatography. ODC activity was measured by the release of (14)CO(2) from (14)C-L-ornithine. Luminal putrescine and cadaverine concentrations were higher in the jejunum than in the ileum, suggesting an absorptive process. The highest concentrations of intestinal polyamines were observed in rats fed the soy protein diet (P < 0.05). Only minor differences were observed in mucosal polyamines according to the diets. ODC activity was also higher in the intestinal mucosa of rats fed the high soy protein diet (P < 0.05). These results suggest that intestinal luminal polyamine concentrations and ODC activity are modulated by the dietary protein source. PMID- 15539317 TI - Beneficial effects of gamma linolenic acid supplementation on nerve conduction velocity, Na+, K+ ATPase activity, and membrane fatty acid composition in sciatic nerve of diabetic rats. AB - Metabolic and vascular abnormalities are implicated in the pathogenesis of diabetic neuropathy. Two principal metabolic defects are altered lipid metabolism resulting from the impairment of delta-6-desaturase, which converts linoleic acid (LA) into gamma linolenic acid (GLA), and reduced nerve Na+, K+ ATPase activity. This reduction may be caused by a lack of incorporation of (n-6) fatty acids in membrane phospholipids. Because this ubiquitous enzyme maintains the membrane electrical potential and allows repolarization, disturbances in its activity can alter the process of nerve conduction velocity (NCV). We studied the effects of supplementation with GLA (260 mg per day) on NCV, fatty acid phospholipid composition, and Na+, K+ ATPase activity in streptozotocin-diabetic rats. Six groups of 10 rats were studied. Two groups served as controls supplemented with GLA or sunflower oil (GLA free). Two groups with different durations of diabetes were studied: 6 weeks with no supplementation and 12 weeks supplemented with sunflower oil. To test the ability of GLA to prevent or reverse the effects of diabetes, two groups of diabetic rats were supplemented with GLA, one group for 12 weeks and one group for 6 weeks, starting 6 weeks after diabetes induction. Diabetes resulted in a 25% decrease in NCV (P < 0.0001), a 45% decrease in Na+, K+ ATPase activity (P < 0.0001), and an abnormal phospholipid fatty acid composition. GLA restored NCV both in the prevention and reversal studies and partially restored Na+, K+ ATPase activity in the preventive treatment group (P < 0.0001). These effects were accompanied by a modification of phospholipid fatty acid composition in nerve membranes. Overall, the results suggest that membrane fatty acid composition plays a direct role in NCV and confirm the beneficial effect of GLA supplementation in diabetic neuropathy. PMID- 15539318 TI - Dietary soy isoflavones inhibit activation of rat platelets. AB - Isoflavones (isoflavonoids) have been proposed to be the active compounds that contribute to decreased mortality from chronic diseases in populations that consume large amounts of soy products. Diets containing soy protein with and without isoflavones were fed to rats to determine if these compounds could exert in vivo effects on physiologic markers of platelet activation. Three methods were employed to monitor platelet activation: measurement of electronic mean platelet volume, which is an indicator of shape change; monitoring of collagen-induced production of reactive oxygen signals (hydrogen peroxide); and determination of increases in phosphorylation of protein tyrosine residues after collagen stimulation. Apparent volumes were significantly smaller for platelets from rats fed isoflavones, suggesting that these platelets were in a more disc-like, quiescent state compared with platelets from rats fed the isoflavone-reduced diet (means +/- SEM, 5.37 +/- 0.08 vs. 5.70 +/- 0.06 fL, n = 6/group, P < 0.008). Results from the other functional tests were consistent with this finding. Platelet production of hydrogen peroxide was found be significantly lower 1, 3, and 5 minutes after addition of collagen for rats fed isoflavones versus rats fed the isoflavone-reduced diet (n = 6/group, P < 0.004). Phosphorylated tyrosine residues in platelet proteins after stimulation also were shown to be significantly lower in the platelets exposed to dietary isoflavones (n = 5/group, P < 0.047). These combined results indicate that soy isoflavones can alter early event signaling networks that result in less activated platelets and may partially explain the beneficial effects of dietary soy against human heart disease. PMID- 15539319 TI - Human peripheral blood mononuclear cells: ; Inhibition of biotin transport by reversible competition with pantothenic acid is quantitatively minor. AB - A transporter present in intestinal cells and in choriocarcinoma cells has been shown to transport both pantothenic acid and biotin at similar transporter affinities. However, the concentration of pantothenic acid in most foods and biological fluids is approximately 200 times the concentration of biotin; theoretically, pantothenic acid might substantially reduce biotin transport via competition. In the present study, we sought to determine whether pantothenic acid reduces biotin transport by the biotin transporter in peripheral blood mononuclear cells (PBMC). PBMC were isolated from human blood by gradient centrifugation. Incubations with [(3)H]biotin and pantothenic acid were conducted at physiologic concentrations. Intracellular [(3)H]biotin was quantified after washing by liquid scintillation counting. Pantothenic acid at 10 to 1,000 nmol/L reduced biotin (475 pmol/L) uptake by less than 12% (P < 0.05). Based on Lineweaver-Burk plots, the competition was reversible. Several structural analogs of pantothenic acid at 1,000 nmol/L reduced biotin transport by only 7 to 15% (P = 0.13). No pattern of molecular structure required for recognition by the transporter was apparent. Extracellular pantothenic acid did not affect biotin efflux from [(3)H]biotin-loaded PBMC (P > 0.05), suggesting that countertransport of extracellular pantothenic acid and intracellular biotin does not increase biotin efflux from PBMC. We conclude that the physiologic effects of pantothenic acid on the transport of biotin in PBMC are likely to be quantitatively minor. PMID- 15539320 TI - Modulating role of glucose on magnesium transport in rat erythrocytes. AB - Magnesium efflux from rat erythrocytes has been shown to be inhibited by a plasma fraction containing glucose. Therefore, we investigated the effect of D-glucose on erythrocyte magnesium transport. We show the inhibitory activity of this hexose on sodium (Na(+))-independent erythrocyte magnesium (Mg(2+)E) efflux. Inhibitory effects of D-mannose, 2-deoxy-D-glucose, and D-fructose on Mg(2+)E efflux also were demonstrated. Moreover, the suppression of the inhibitory activity of glucose on Mg(2+)E efflux was shown to be associated with the inhibition of glucose transport by cytochalasin B and phloretin. Together these data suggest a possible implication of the glucose carrier GLUT-1 in the regulation of Mg(2+) transport. PMID- 15539321 TI - Experimental studies on antiarrhythmic and antiseizure effects of polyunsaturated fatty acids in excitable tissues. AB - It has been shown that in animals, and probably in humans, n-3 polyunsaturated fatty acids (PUFAs) are antiarrhythmic. We discuss our recent studies on the antiarrhythmic actions of PUFAs. PUFAs stabilize the electrical activity of isolated cardiac myocytes by requiring a stronger electrical stimulus to elicit an action potential and by markedly prolonging the refractory period. These electrophysiologic effects are the result of specific modulation of ion currents, particularly of the voltage-dependent sodium current and of the L-type calcium currents across sarcolemmal phospholipid membranes. This appears to be the probable major antiarrhythmic mechanism of PUFAs. However, they also similarly affect neuronal ion channels with potentially important functional effects on the nervous system. PMID- 15539322 TI - Availability and antiperoxidative effects of beta-carotene from Dunaliella bardawil in alcohol-drinking rats. AB - The present study demonstrated the high bioavailability and antiperoxidative capacity of the natural beta-carotene isomer mixture of Dunaliella bardawil compared with synthetic beta-carotene under alcohol-induced oxidative stress. Weanling rats were adapted to ethanol by increasing ethanol levels in their drinking water to 30% at 5% intervals per week; other rats received water with no added ethanol. One water-drinking group and one alcohol-drinking group with no dietary carotene were used as controls. Two water-drinking groups were supplemented with 1 g/kg diet beta-carotene either from Dunaliella or a synthetic source, and due to reduced food intake, two ethanol-fed groups received 2 g beta carotene per kilogram of diet from each source. Following 3 months of ethanol consumption, both carotene sources were found to prevent ethanol-induced lipid peroxidation as expressed by the hepatic conjugated oxidized dienes level. However, in the algal-fed rats, hepatic carotene and vitamin A levels were higher. In addition to a lower performance of the group fed ethanol and synthetic beta-carotene, there were three deaths in this group. PMID- 15539323 TI - Lipotropes alter casein gene expression in bovine mammary acinar culture. AB - Lipotropes (methyl group containing nutrients, including methionine and choline, folic acid, and vitamin B(12)) are essential for cell proliferation and differentiation of mammary tissues. Lipotropes interact in the supply and regulation of intracellular methyl group pools, thereby affecting synthesis and methylation of DNA. To determine the effect of lipotropes on milk protein gene expression, acini isolated from mammary tissues of lactating cows were cultured in one of three treatment media: (1) control, (2) lipotrope deficient, and (3) lipotrope supplemented. beta-Casein mRNA was determined by Northern blotting, and milk protein secretion was measured by a pulse-chase method. The level of beta casein mRNA was lower in cells grown in lipotrope-deficient medium than in cells grown in the lipotrope-supplemented and control media. Acinar cells cultured in lipotrope-deficient medium also had approximately threefold less milk protein secretion than that of cells in either control or lipotrope-supplemented media. Protein secretion did not differ in the control and lipotrope-supplemented groups. The present study indicates that lipotrope deficiency suppresses total protein secretion and beta-casein gene expression in bovine mammary alveolar epithelial cells in culture. PMID- 15539324 TI - Acute ingestion of different dietary fatty acid species modulates postprandial lipid responses in New Zealand white rabbits. AB - Although several investigations have linked the degree of fatty acid saturation to plasma lipid responses in the postprandial state, further evaluation is necessary. In this study, we compared the effect of saturated (SFA), monounsaturated (MUFA), and polyunsaturated (PUFA) fatty acids on postprandial lipid metabolism using complementary in vivo and in vitro approaches. Fat (10 g) cholesterol (0.5 g) test meals that provided either lard (SFA), olive oil (MUFA), or sunflower oil (PUFA) were ingested by chow-fed New Zealand white rabbits (n = 8). In addition, hepatic uptake of triglyceride-cholesterol-rich lipoproteins (TCRL) isolated from rabbits chronically ingesting SFA, MUFA, or PUFA diets was measured using freshly isolated chow-fed rabbit hepatocytes. Whatever dietary fatty acids ingested, postprandial triglyceridemia and occurrence of radiolabelled dietary lipids in plasma were not markedly different. Conversely, SFA induced higher postprandial cholesterolemia and phospholipemia than MUFA (P < 0.05) whereas PUFA prevented postprandial cholesterol increase. TCRL disappearance from cultured liver cell media was delayed with SFA-rich TCRL and faster with PUFA whereas MUFA-rich TCRL showed an intermediate figure. From these data, we conclude that SFA, MUFA, and PUFA elicited different postprandial plasma and lipoprotein lipid responses. The fatty acid composition of TCRL had a major impact on their subsequent metabolism, especially uptake by cultured hepatocytes. The SFA-induced hypercholesterolemia could be related to an altered hepatic uptake whereas a faster clearance and hepatic uptake could explain the cholesterol-lowering effect of PUFA in rabbits. MUFA, like PUFA, accelerate uptake by hepatocytes but favor cholesterol ester enrichment of TCRL. PMID- 15539325 TI - Effect of vitamin B6 supplementation on degradation rates of short-lived proteins in human neutrophils. AB - Metabolic pathways are controlled primarily by protein degradation rates. Degradation rates, in turn, are controlled by changes in physiologic condition or nutrient supply. Vitamin B(6) is associated with a greater variety of reactions than most other vitamins. Moreover, the vitamin B(6) needs of the elderly tend to be higher than those of young adults. Neutrophils seem to be appropriate cells for assessing protein turnover as affected by macronutrients and micronutrients. Thus, we assumed that vitamin B(6) supplementation, particularly in an elderly population, would change the turnover rates of the neutrophil proteins. Protein synthesis was measured after 30 minutes of (35)S-Met incorporation followed by a 30-minute washout incubation; degradation was measured after an additional 5-hour incubation. Following protein separation, radioactive images of short-lived proteins were electronically separated into bands. Vitamin B(6) supplementation significantly increased the synthesis of most neutrophil protein bands. There was a significant decrease of 25 to 66% in the degradation rates of 235 protein bands. We even detected by statistical evaluation a 20% decrease in the degradation rates of distinct protein bands. Activation coefficients of erythrocyte aspartate aminotransferase (AC-AST) decreased markedly. There was a significant positive correlation between the decrease in AC-AST and protein degradation. The N-end rule proposes that pyridoxal 5'-phosphate decreases degradation rates of short-lived proteins by binding to lysyl residues. A biochemical model of the mechanism of cellular protein turnover, as affected by nutritional intervention, in human neutrophils is demonstrated. PMID- 15539326 TI - Renal lesions in streptozotocin-induced diabetic rats maintained on onion and capsaicin containing diets. AB - Onion (Allium cepa) powder and capsaicin, the pungent principle of red pepper (Capsicum annum) were added in the amounts of 3 g% and 15 mg%, respectively, to the diet of streptozotocin-induced diabetic Wistar rats for 8 weeks. The presence of renal lesions was assessed by the extent and quality of proteinuria and by the leaching of renal tubular enzymes into the urine. Renal integrity was assessed by measuring the activities in the kidney tissue of several key enzymes of carbohydrate metabolism and of polyol pathway, transaminases, and ATPases. Data on enzymuria and proteinuria, activities of kidney ATPases present in diabetic patients, suggested that dietary onion caused significant beneficial modulation of the progression of renal lesions in the diabetic rats. These findings were also corroborated by histologic examination of kidney sections. Dietary capsaicin did not have any favorable influence on renal pathology in diabetes. It is inferred that this beneficial ameliorating influence of dietary onion on diabetic nephropathy may be mediated through onion's ability to lower blood cholesterol levels and to reduce lipid peroxidation. PMID- 15539327 TI - The influence of zinc status on the kinetics of zinc uptake into cultured endothelial cells. AB - To better understand cellular zinc homeostasis and characterize the zinc transport process, a mammalian cell culture model was utilized to investigate the influence of zinc status on the kinetics of zinc uptake. Culturing conditions were optimized to induce moderate zinc deficiency and zinc excess while still sustaining the general health of the cells. Cells were grown in (1) control medium of 10% fetal bovine serum (FBS) in minimum essential medium (MEM; 5.0 micromol zinc/L), (2) low zinc medium (10% dialyzed FBS in MEM; 1.5 micromol zinc/L), or (3) zinc back medium (10% dialyzed FBS in MEM with zinc added as ZnCl(2); 5.0 micromol zinc/L). Bovine pulmonary artery endothelial cells (BPAEC), porcine aortic endothelial cells (PAEC), and porcine venous endothelial cells (PVEC) were evaluated as to their responsiveness to our zinc-deficient conditions. Zinc uptake was faster (P < 0.001) in all three cell types when they were grown in low zinc medium compared with controls; the increases were 32% in PAEC, 37% in PVEC, and 66% in BPAEC. Further kinetic analysis with BPAEC demonstrated a 31% increase (P < 0.05) in the maximum rate of zinc uptake (Jmax) grown in low zinc medium compared with controls, but no difference (P > 0.05) between the low zinc group and the control group in the concentration at which uptake was half-maximal (K). Zinc uptake into BPAEC grown in excess zinc conditions was not different (P > 0.05) unless the medium contained greater than 50 micromol zinc/L. In conclusion, BPAEC increased their ability for zinc uptake in response to moderate zinc deficiency, but did not change their kinetics of zinc uptake during moderate zinc excess. PMID- 15539328 TI - A new HPLC method for the simultaneous determination of oxidized and reduced plasma aminothiols using coulometric electrochemical detection. AB - A new method has been developed that is capable of providing a complete profile of the most common monothiols and disulfides present in plasma or tissue extracts. The method utilizes reversed phase ion-pairing high performance liquid chromatography coupled with coulometric electrochemical detection to simultaneously quantify free oxidized and reduced aminothiols or total aminothiols after chemical reduction. The method is extremely sensitive, with limits of detection in the 5 fmol/mL range for monothiols and 50 fmol/mL for dithiols. The interassay and intraassay coefficients of variation for total and free aminothiols ranged between 1.2 and 5.8%. The mean recoveries for total and plasma aminothiols ranged between 97.1 and 102.8%. The aminothiols are quantified directly, without derivatization, and include methionine, homocysteine, homocystine, cystathionine, cysteine, cystine, cysteinylglycine, and oxidized and reduced glutathione. Because a complete aminothiol profile of metabolites in both the remethylation (anabolic) and transulfuration (catabolic) pathways of homocysteine metabolism can be determined simultaneously, this new method should be useful in determining the metabolic etiology of homocysteinemia and in designing appropriate nutritional intervention strategies. Basic research applications of this method should lead to an increased understanding of the metabolic pathology of aminothiol imbalance. PMID- 15539329 TI - Effects of soluble corn bran arabinoxylans on cecal digestion, lipid metabolism, and mineral balance (Ca, Mg) in rats. AB - The effects of soluble corn bran arabinoxylans on cecal digestion, lipid metabolism, and mineral utilization [calcium (Ca) and magnesium (Mg)] were investigated in rats adapted to semipurified diets. The diets provided either 710 g/kg wheat starch alone (control) or 610 g/kg wheat starch plus 100 g/kg corn soluble fiber (arabinoxylans) and either 0 or 2 g/kg cholesterol (control + cholesterol and arabinoxylans + cholesterol, respectively). Compared with rats fed the control diets, rats fed the arabinoxylan diets had significant cecal hypertrophy (+50% after 3 days of the fiber adaptation) and an accumulation of short-chain fatty acids, especially propionic acid (up to 45% in molar percentage). Arabinoxylans enhanced the cecal absorption of Ca and Mg (from 0.07 to 0.19 micromol/min for Ca and from 0.05 to 0.23 micromol/min for Mg). Mg balance was enhanced by arabinoxylans (+25%). The arabinoxylan diet markedly reduced the cholesterol absorption from 50% of ingested cholesterol in controls up to approximately 15% in rats adapted to the arabinoxylans diet. Arabinoxylans were effective in lowering plasma cholesterol (approximately -20%). There was practically no effect of the diets on cholesterol in d > 1.040 lipoproteins (high density lipoproteins) whereas arabinoxylans were very effective in depressing cholesterol in d < 1.040 lipoproteins (especially in triglyceride-rich lipoproteins). Corn fermentable fiber decreased the accumulation of cholesterol in the liver. In parallel, the arabinoxylan diet counteracted the downregulation of 3-hydroxy-3-methylglutaryl-CoA by cholesterol. These data suggest that arabinoxylans may have a great impact on intestinal fermentation, mineral utilization, and cholesterol metabolism. PMID- 15539330 TI - Effects of the dietary phytoestrogen biochanin A on cell growth in the mammary carcinoma cell line MCF-7. AB - Studies of the dietary phytoestrogen biochanin A on cell proliferation of the cultured estrogen responsive cells human breast carcinoma MCF-7 showed that biochanin A exhibits biphasic regulation on MCF-7 cells. At concentrations of less than 10 microg/mL, cells respond to biochanin A by increasing cell growth and de novo DNA synthesis. The addition of biochanin A at concentrations of greater than 30 microg/mL significantly inhibited cell growth and DNA synthesis in a dose-dependent fashion, resulting in an IC(50) value of 40 microg/mL. The reversibility of these inhibitory effects by biochanin A appears also to be concentration dependent. Cells previously treated with high concentrations (>60 microg/mL) of biochanin A did not regain normal growth after treatment ceased. Biochanin A was cytostatic at low concentrations (<40 microg/mL) and cytotoxic at higher concentrations. Upon exposure to 100 microg/mL of biochanin A, cell morphology was severely altered, cell volume decreased, and condensation of cell components was clearly noticeable. In addition, biochanin A damaged cell membranes by increasing membrane permeability. These results suggest possible molecular and cellular mechanisms of the action of dietary phytoestrogens on estrogen target cells. PMID- 15539331 TI - Regulation of polyamine synthesis and transport by retinoic acid and epidermal growth factor in cultured adult rat type II pneumocytes. AB - During injury of lung epithelial cells, the type II pneumocyte proliferates and differentiates into a type I pneumocyte to restore the epithelium. Polyamines, which constitute a family of small organic polycations, are required for this process of cell repair. Because retinoic acid (RA) and epidermal growth factor (EGF) also are involved, the purpose of this research was to determine their effect on polyamine transport and synthesis in cultured type II pneumocytes. Rat type II pneumocytes were isolated, cultured overnight, and treated with RA and/or EGF for 24 hours. Polyamine transport was determined by [(3)H]spermidine uptake, and polyamine synthesis was assessed by the activity of the initial rate-limiting enzyme ornithine decarboxylase. EGF (100 ng/mL) significantly increased spermidine transport, but RA did not. At low concentrations of spermidine (2 microM), the combined effect of RA and EGF on spermidine transport was additive. Both EGF (25 ng/mL) and RA (1 microM) increased polyamine synthesis, and cotreatment resulted in an additive effect (a fourfold increase over the control). We also found that ornithine decarboxylase activity is greatly diminished in the presence of tyrphostin B56, which is a specific inhibitor for the tyrosine kinase of the EGF receptor, suggesting that polyamine synthesis within the type II pneumocyte may depend on activation of tyrosine kinase of the EGF receptor. These results indicate that RA and EGF increase the availability of polyamines, which may be important in the lung cell repair process. PMID- 15539332 TI - Vitamin B6 deficiency decreases the glucose utilization in cognitive brain structures of rats. AB - The effects of vitamin B(6) deficiency on metabolic activities of brain structures were studied. Male Sprague-Dawley weanling rats received one of the following diets: (1) 7 mg pyridoxine HCl/kg (control group); (2) 0 mg pyridoxine HCl/kg (vitamin B(6)-deficient group); or (3) 7 mg pyridoxine HCl/kg with food intake restricted in quantity to that consumed by the deficient group (pair-fed control group). After 8 weeks of dietary treatment, rats in all three groups received an intravenous injection of 2-deoxy-[(14)C] glucose (100 microCi/kg). Vitamin B(6) status was evaluated by plasma pyridoxal 5'-phosphate concentrations. The vitamin B(6)-deficient group had significantly lower levels of plasma pyridoxal 5'-phosphate than did the control and pair-fed groups. The local cerebral glucose utilization rates in structures of the limbic system, basal ganglia, sensory motor system, and hypothalamic system were determined. The local cerebral glucose utilization rates in each of the four brain regions in the deficient animals were approximately 50% lower (P < 0.05) than in the control group. Results of the present study suggest that serious cognitive deficit may occur in vitamin B(6)-deficient animals. PMID- 15539333 TI - Short-term beta-carotene supplementation of lactating mothers consuming diets low in vitamin A. AB - We have previously shown that beta-carotene supplementation of the diets of healthy U.S. mothers increases serum and milk beta-carotene concentrations. Building on these results, we investigated the possibility that beta-carotene supplementation could enhance the vitamin A status of mothers and their nursing infants. Three 30-mg doses of beta-carotene were administered on 3 consecutive days to 44 lactating mothers who had vitamin-A-poor diets. Concentrations of maternal serum and milk carotenoids and retinol were evaluated at baseline and after 2 and 3 days of supplementation. Infant serum carotenoids and retinol were measured at baseline and 2 days following maternal supplementation. beta-Carotene supplementation markedly elevated maternal serum and milk beta-carotene concentrations (nine- and sevenfold, respectively) and resulted in smaller, transient increases of alpha-carotene, lycopene, and beta-cryptoxanthin concentrations in maternal serum. Maternal serum and milk retinol were unchanged in response to the treatment. In contrast, maternal beta-carotene supplementation significantly increased infant serum retinol (P 0.05) or following drug infusion (SNP: 1.36 +/- 0.21; ISO: 1.27 +/- 0.23 AU mmHg( 1), P > 0.05). The reduction in CVC during the subsequent cold stress was significantly less at the SNP site (38.1 +/- 6.2%) relative to the ISO site (65.0 +/- 5.5%; P= 0.007). These data suggest NO is capable of inhibiting sympathetically mediated vasoconstriction in the cutaneous vasculature. PMID- 15539402 TI - Preceding muscle activity influences motor unit discharge and rate of torque development during ballistic contractions in humans. AB - To investigate the effect of initial conditions on the modulation of motor unit discharge during fast voluntary contractions, we compared ballistic isometric contractions of the ankle dorsiflexor muscles that were produced from either a resting state or superimposed on a sustained contraction. The torque of the dorsiflexors and the surface and intramuscular EMGs from the tibialis anterior were recorded. The results showed that the performance of a ballistic contraction from a sustained contraction ( approximately 25% maximal voluntary contraction (MVC)) had a negative effect on the maximal rate of torque development. Although the electromechanical delay was shortened, the EMG activity during the ballistic contraction was less synchronized. These observations were associated with a significant decline in the average discharge rate of single motor units (89.8 +/- 3.8 versus 115 +/- 5.8 Hz) and in the percentage of units (6.2 versus 15.5% of the whole sample) that exhibited double discharges at brief intervals (= 5 ms). High-threshold units that were not recruited during the sustained contraction displayed the same activation pattern, which indicates that the mechanisms responsible for the decline in discharge rate were not restricted to previously activated units, but appear to influence the entire motor unit pool. When a premotor silent period (SP) was observed at the transition from the sustained muscular activity to the ballistic contraction (19% of the trials), these adjustments in motor unit activity were not present, and the ballistic contractions were similar to those performed from a resting state. Together, these results indicate that initial conditions can influence the capacity for motor unit discharge rate and hence the performance of a fast voluntary contraction. PMID- 15539404 TI - Down-regulation of the receptor for advanced glycation end-products (RAGE) supports non-small cell lung carcinoma. AB - The receptor for advanced glycation end-products (RAGE) is a transmembrane receptor of the immunoglobulin superfamily. Several ligands binding to RAGE have been identified, including amphoterin. Experimental studies have given rise to the discussion that RAGE and its interaction with amphoterin contribute to tumour growth and metastasis. However, none of the studies considered a differential transcription profile in cancer that might change the interpretation of the study results when comparing RAGE in tumours with histologically normal tissues. Here we show that RAGE is strongly reduced at the mRNA and even more so at the protein level in non-small cell lung carcinomas compared with normal lung tissues. Down regulation of RAGE correlates with higher tumour (TNM) stages but does not depend on the histological subtypes, squamous cell lung carcinoma and adenocarcinoma. Subsequent overexpression of full-length human RAGE in lung cancer cells (NCI H358) showed diminished tumour growth under some conditions. While proliferation of RAGE-expressing cells was less than that of cells expressing the cytoplasmic domain deletion mutant DeltacytoRAGE or mock-transfected NCI-H358 in monolayer cultures, RAGE cells also formed smaller tumours in spheroid cultures and in vivo in athymic mice compared with DeltacytoRAGE cells. Moreover, we observed a more epithelial growth of RAGE-expressing, but also of DeltacytoRAGE-expressing, cells on collagen layers, whereas mock NCI-H358 cells kept their tumour morphology. This observation was supported by immunofluorescence analyses demonstrating that RAGE preferentially localizes at intercellular contact sites, independent of expression of the cytoplasmic domain. Thus, down-regulation of RAGE may be considered as a critical step in tissue reorganization and the formation of lung tumours. PMID- 15539403 TI - Modulation of body fluids and angiotensin II receptors in a rat model of intra uterine growth restriction. AB - We previously reported that sodium restriction during pregnancy reduces plasma volume expansion and promotes intra-uterine growth restriction (IUGR) in rats while it activates the renin-angiotensin-aldosterone system (RAAS). In the present study, we proceeded to determine whether expression of the two angiotensin II (ANGII) receptor subtypes (AT(1) and AT(2)) change in relation to maternal water-electrolyte homeostasis and fetal growth. To this end, pregnant (gestation day 15) and non-pregnant Sprague-Dawley rats were randomly assigned to two groups fed either normal, or Na(+)-restricted diets for 7 days. At the end of the treatment period, plasma aldosterone and renin activity as well as plasma and urine electrolytes were measured. Determinations for AT(1) and AT(2) mRNA and protein were made by RNase protection assay and photoaffinity labelling, respectively, using a number of tissues implicated in volume regulation and fetal growth. In non-pregnant rats, Na(+) restriction decreases Na(+) excretion without altering plasma volume, plasma Na(+) concentration or the expression of AT(1) and AT(2) mRNA or protein in the tissues examined. In normally fed pregnant rats when compared to non-pregnant controls, AT(1) mRNA increases in the hypothalamus as well as pituitary and declines in uterine arteries, while AT(1) protein decreases in the kidney and AT(2) mRNA declines in the adrenal cortex. In pregnant rats, Na(+) restriction induces a decrease in plasma Na(+), an increase in plasma urea, as well as a decline in renal urea and creatinine clearance rates. Protein levels for both AT(1) and AT(2) in the pituitary and AT(2) mRNA in the adrenal cortex are lower in the Na(+)-restricted pregnant group when compared to normally fed pregnant animals. Na(+) restriction also induces a decrease in AT(1) protein in the placenta. In conclusion, these results suggest that pregnancy may increase sensitivity to Na(+) depletion by the tissue-specific modulation of ANGII receptors. Finally, these receptors may be implicated in the IUGR response to low Na(+). PMID- 15539405 TI - 1 alpha,25-Dihydroxyvitamin D(3) is a preventive factor in the metastasis of lung cancer. AB - 1alpha,25-Dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], the major regulator of calcium homeostasis, has potent antiproliferative and anti-invasive properties in vitro in cancer cells. Studies in vivo demonstrated that 1alpha,25(OH)(2)D(3) slows the progression of breast, prostate and other carcinomas. A key question is whether 1alpha,25(OH)(2)D(3) exerts its anticarcinogenic effects in vivo by a mechanism that is dependent on its capacity to limit the proliferation and invasiveness of cancer cells in vitro. It has not been clear whether the calcemic activity and regulation of the host defenses by 1alpha,25(OH)(2)D(3) contribute to the effect on cancer cells. In this study we have focused on the influence of 1alpha,25(OH)(2)D(3) on the metastasis of lung cancer, without involvement of the calcemic activity and other effects of 1alpha,25(OH)(2)D(3) in the host. We used metastatic Lewis lung carcinoma cells expressing green fluorescent protein (LLC GFP cells) and examined metastatic activity in vitamin D receptor (VDR) null mutant (VDR(-/-)) mice and their wild-type counterparts (VDR(+/+) mice). VDR(-/-) mice exhibit hypocalcemia and extremely high serum levels of 1alpha,25(OH)(2)D(3). We expected that serum 1alpha,25(OH)(2)D(3) would act in vivo to directly inhibit the metastatic growth of VDR-positive LLC-GFP cells in VDR(-/-) mice. The metastatic activities of LLC-GFP cells were remarkably reduced in VDR(-/-) mice compared with VDR(+/+) mice. To test the hypothesis that serum 1alpha,25(OH)(2)D(3) is an intrinsic factor that inhibits metastatic growth of lung cancer cells, we corrected hypocalcemia and/or hypervitaminosis D in VDR(-/ ) mice by dietary manipulation. The metastatic growth of LLC-GFP cells was remarkably reduced in response to serum levels of 1alpha,25(OH)(2)D(3), but not to serum calcium levels. Furthermore, we found that VDR(+/+) mice fed the manipulated diets displayed an apparent inverse relationship between the physiological levels of serum 1alpha,25(OH)(2)D(3) (8-15 pg/ml) and tumorigenesis. Here we show that 1alpha,25(OH)(2)D(3) inhibits the metastatic growth of lung cancer cells in a defined animal model. PMID- 15539406 TI - Differential gene expression in rat colon by dietary heme and calcium. AB - Dietary heme and calcium are alleged modulators of colon cancer risk. Little is known about the molecular and cellular changes in the colon epithelium that are induced by consumption of these unabsorbed nutrients. In this nutrigenomics study, we fed rats high- and low-calcium diets with or without heme. In agreement with previous studies, we found that dietary heme increased the cytotoxicity of fecal water in the colon and elevated epithelial proliferation, a risk factor in colon carcinogenesis. Calcium reduced cytotoxicity and inhibits heme-induced effects. Among 365 colon-expressed genes, we could identify 10 diet-modulated genes that show >2-fold altered expression, of which several are related to colon cell turnover and disease. Mucosal pentraxin (Mptx) was the strongest differentially expressed gene, approximately 10-fold down-regulated by dietary heme and 3-fold up-regulated by calcium. cDNA microarray and quantitative PCR analysis show that calcium significantly inhibits the effects of heme, which correlates with the physiological effects. Our results indicate that Mptx expression is related to colonic cell turnover, and that Mptx might be a marker for diet-modulated mucosal integrity. We also show that Mptx expression is restricted to the intestine, and occurs predominantly in the colon. PMID- 15539407 TI - Regulated association of protein kinase B/Akt with breast tumor kinase. AB - Increased protein-tyrosine kinase activity is a prognostic indicator of decreased disease-free survival in patients with advanced breast tumors. Breast tumor kinase (Brk) is a soluble protein-tyrosine kinase overexpressed in the majority of breast cancers and also in normal skin and gut epithelium, but not in normal breast epithelial cells. Herein, we show that Brk interacts with protein kinase B/Akt, a serine/threonine kinase involved in cell growth and survival. Epidermal growth factor (EGF) treatment of human keratinocytes or Brk-transfected COS-1 cells leads to the dissociation of the Brk.Akt complex, whereas a constitutively active Brk mutant containing a point mutation at Tyr-447 (YF-Brk) failed to dissociate from Akt upon EGF treatment. In addition, Brk.Akt dissociation was blocked by the inhibition of phosphatidylinositol 3-kinase. Similar to ectopic Brk, endogenous Brk in T47D breast cancer cells was less phosphorylated upon EGF treatment, but it remained constitutively associated with Akt in the presence of EGF. Overexpression of wild-type (wt)-Brk, kinase-inactive (KM)-Brk, or YF-Brk increased the Tyr phosphorylation of multiple signaling molecules including EGF receptor. However, only wt- and YF-Brk, but not KM-Brk, induced phosphorylation of Akt and inhibited the kinase activity of Akt in unstimulated cells. Similarly, overexpression of wt- or YF-, but not KM-Brk, blocked the phosphorylation of the forkhead transcription factor, a downstream Akt target. These results suggest that Brk may function as a signaling molecule whose kinase activity normally limits the activity of Akt in unstimulated cells. Additionally, these results suggest that in breast cancer cells Brk behaves similarly to a constitutively active Brk mutant (YF-Brk) and associates with tyrosine-phosphorylated proteins in deregulated signaling complexes. Together these data provide clues to the possible proto-oncogenic and oncogenic functions of Brk. PMID- 15539408 TI - Structures of dCTP deaminase from Escherichia coli with bound substrate and product: reaction mechanism and determinants of mono- and bifunctionality for a family of enzymes. AB - dCTP deaminase (EC 3.5.4.13) catalyzes the deamination of dCTP forming dUTP that via dUTPase is the main pathway providing substrate for thymidylate synthase in Escherichia coli and Salmonella typhimurium. dCTP deaminase is unique among nucleoside and nucleotide deaminases as it functions without aid from a catalytic metal ion that facilitates preparation of a water molecule for nucleophilic attack on the substrate. Two active site amino acid residues, Arg(115) and Glu(138), were identified by mutational analysis as important for activity in E. coli dCTP deaminase. None of the mutant enzymes R115A, E138A, or E138Q had any detectable activity but circular dichroism spectra for all mutant enzymes were similar to wild type suggesting that the overall structure was not changed. The crystal structures of wild-type E. coli dCTP deaminase and the E138A mutant enzyme have been determined in complex with dUTP and Mg(2+), and the mutant enzyme also with the substrate dCTP and Mg(2+). The enzyme is a third member of the family of the structurally related trimeric dUTPases and the bifunctional dCTP deaminase-dUTPase from Methanocaldococcus jannaschii. However, the C terminal fold is completely different from dUTPases resulting in an active site built from residues from two of the trimer subunits, and not from three subunits as in dUTPases. The nucleotides are well defined as well as Mg(2+) that is tridentately coordinated to the nucleotide phosphate chains. We suggest a catalytic mechanism for the dCTP deaminase and identify structural differences to dUTPases that prevent hydrolysis of the dCTP triphosphate. PMID- 15539409 TI - Role of hepatocyte nuclear factors in transcriptional regulation of male-specific CYP2A2. AB - Cytochrome P450 2A2 (CYP2A2) is an adult male-specific rat liver steroid hydroxylase whose sex-dependent expression is regulated at the transcriptional level by sexually dimorphic pituitary growth hormone (GH) secretory patterns. In contrast to CYP2C11 and other male-specific, plasma GH pulse-inducible liver genes, CYP2A2 is highly expressed in hypophysectomized rat liver, despite the absence of GH stimulation. CYP2A2 promoter fragments 0.9-6.2 kb long exhibited unusually high basal promoter activity when transfected into the liver cell line HepG2. A further approximately 2.5-fold increase in activity was obtained by cotransfection of hepatocyte nuclear factor (HNF) 3gamma or HNF4alpha. CYP2A2 promoter activity was inhibited approximately 85% by transfection of HNF3beta or HNF6, both of which are more highly expressed in female than male liver and can strongly trans-activate the female-specific CYP2C12 promoter. The male GH pulse activated transcription factor STAT5b had no effect on CYP2A2 promoter activity, either alone or in combination with HNF3gamma and HNF4alpha, consistent with the GH pulse-independence of CYP2A2 expression. By contrast, STAT5b synergistically enhanced the transcriptional activity of HNF4alpha toward two other male-specific liver target genes, Cyp2d9 and CYP8B1. Furthermore, STAT5b in combination with the HNF4alpha coactivator peroxisome proliferator-activated receptor gamma coactivator-1alpha strongly enhanced the transcriptional activity of HNF4alpha toward CYP8B1 but not toward CYP2A2. These findings support the hypothesis that sex-dependent HNFs contribute to the sexually dimorphic expression of CYP2A2 and other liver CYPs and highlight the ability of STAT5b to act in concert with HNF4alpha to regulate select male-specific liver CYP genes. PMID- 15539410 TI - Pregnancy in patients with rheumatic disease: anti-inflammatory cytokines increase in pregnancy and decrease post partum. AB - OBJECTIVE: To investigate changes in the levels of circulating cytokines with a focus on the Th1/Th2 balance during and after pregnancy in patients with rheumatoid arthritis (RA), juvenile idiopathic arthritis (JIA), and ankylosing spondylitis (AS). METHODS: Plasma and serum samples of 34 pregnant patients, 19 with RA, 6 with JIA, and 9 with AS, and of 30 healthy pregnant women, 20 non pregnant patients, and 10 non-pregnant healthy women were analysed for levels of interferon gamma (IFNgamma), interleukin (IL) 1beta, IL10, IL1 receptor antagonist (IL1Ra), soluble tumour necrosis factor receptor (sTNFR), and soluble CD30 (sCD30) by ELISA. Clinical assessment and blood sampling in pregnant women was done once in each trimester and 6, 12, and 24 weeks post partum. Disease activity in the patients was evaluated by validated clinical instruments and correlated with circulating levels of cytokines. RESULTS: Low levels of IL10 were found sporadically, whereas IFNgamma and IL1beta were below detection level in the samples tested. Significantly higher concentrations of sTNFR and IL1Ra were measured in pregnant than in non-pregnant subjects. An increase of IL1Ra from the second to the third trimester correlated with improvement of disease activity in patients with RA and AS. Compared with non-pregnant patients and the other pregnant women, patients with RA showed markedly raised levels of sCD30 during pregnancy. CONCLUSIONS: IFNgamma and IL10, markers of a Th1 and Th2 response, respectively, were either low or undetectable in the cohorts analysed. The increase of cytokine inhibitors IL1Ra and sTNFR was related to pregnancy and was independent of an underlying disease. These anti-inflammatory mediators seem to affect disease activity. PMID- 15539411 TI - Lack of genetic association of the three more common polymorphisms of CARD15 with psoriatic arthritis and psoriasis in a German cohort. AB - OBJECTIVE: To determine whether the three common independent sequence variants of the putative pleiotropic non-MHC autoimmune gene CARD15 influence disease susceptibility in large German cohorts of patients with psoriatic arthritis and psoriasis vulgaris, before and after stratification to HLA-C. METHODS: DNA was obtained from 375 patients with psoriatic arthritis, 281 patients with psoriasis vulgaris without joint involvement, and 376 controls. The three variants of the CARD15 gene (R702W, G908R, leu1007fsinsC), and two single nucleotide polymorphisms of the HCR gene (HCR-325, HCR-2327) for HLA-C stratification were genotyped using allelic discrimination Taqman assays. RESULTS: No significant differences in genotype frequencies were observed between controls and either the psoriatic arthritis or the psoriasis vulgaris patient population, even after stratification to HLA-C in both patient cohorts, or to the type of joint involvement within the psoriatic arthritis group. CONCLUSIONS: The lack of genetic association between the most common Crohn's disease alleles of the CARD15 gene and psoriatic joint disease on large cohorts of white patients does not support a recently claimed role for CARD15 as the first non-MHC susceptibility gene in the pathogenesis of psoriatic arthritis, but confirms and extends previous studies in the case of psoriasis vulgaris. PMID- 15539412 TI - Depletion of synovial macrophages in rheumatoid arthritis by an anti-FcgammaRI calicheamicin immunoconjugate. AB - BACKGROUND: Monocytes/macrophages have an important and versatile role in joint inflammation and destruction in rheumatoid arthritis (RA). OBJECTIVE: To determine the efficiency of monocyte/macrophage elimination by a new drug conjugated antibody (CD64-calicheamicin (CD64-CaMi)) directed to the high affinity receptor for IgG (FcgammaRI). METHODS: Mononuclear cells from peripheral blood and synovial fluid of patients with RA were cultured in the presence of CD64-CaMi. Cell death of monocytes/macrophages was measured by analysis of phenotypic changes (light scatter patterns, CD14 expression, and FcgammaRI expression) and nuclear DNA fragmentation. The selectivity of CD64-CaMi was checked by using FcgammaRI deficient and FcgammaRI transfected cell lines. In addition, the indirect effect of CD64-CaMi-induced macrophage cell death on arthritogenic T(h1) cell activity was determined. RESULTS: Inflammatory macrophages from RA synovial fluid, expressing increased FcgammaRI levels, were efficiently killed by CD64-CaMi through induction of DNA fragmentation. CD64-CaMi induced cell death of monocytes/macrophages from peripheral blood of patients with RA proved less efficient. Induction of synovial macrophage death by CD64 CaMi was accompanied by efficient inhibition of proinflammatory T(h1) cytokine production. CONCLUSION: Together, the presented data suggest that elimination of macrophages through a new FcgammaRI directed CD64-CaMi is feasible. Because monocytes from peripheral blood are also eliminated by this immunoconjugate, additional experimental studies should validate its potential for local (intra articular) application in the treatment of RA. PMID- 15539413 TI - CARD15 gene polymorphisms in patients with spondyloarthropathies identify a specific phenotype previously related to Crohn's disease. AB - BACKGROUND: The association between spondyloarthropathy and Crohn's disease is well known. A risk for evolution to Crohn's disease has already been shown in the subgroup of patients with spondyloarthropathy associated with chronic gut inflammation. OBJECTIVE: To investigate whether the reported polymorphisms in the CARD15 gene, a susceptibility gene for Crohn's disease, are associated with the presence of preclinical intestinal inflammation observed in spondyloarthropathies. METHODS: 104 patients with spondyloarthropathies were studied. All underwent ileocolonoscopy with biopsies between 1983 and 2004. The prevalence of three single nucleotide polymorphisms in the CARD15 gene (R702W, G908R, and 1007fs) was assessed using restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR); the patients were compared with an ethnically matched Crohn's disease population and a control population. RESULTS: The carrier frequency of R702W, G908R, or 1007fs variants in the spondyloarthropathy populations (20%) was similar to the control population (17%), but increased to 38% in the spondyloarthropathy subgroup with chronic gut inflammation. This frequency was significantly higher than in the other spondyloarthropathy subgroups (p = 0.001) or the control group (p = 0.006), but not different from the Crohn's disease group (49%) (NS). This indicates that CARD15 polymorphisms are associated with a higher risk for development of chronic gut inflammation. CONCLUSIONS: CARD15 gene polymorphisms clearly identify a subgroup of patients with spondyloarthropathies associated with chronic intestinal inflammation. PMID- 15539414 TI - Lumbosacral radiographic signs in patients with previous enteroarthritis or uroarthritis. AB - OBJECTIVE: To investigate if there are differences in radiological findings in lumbosacral spine radiographs between different subgroups of patients with previous reactive arthritis. METHODS: 95 patients with previous enteroarthritis (n = 53), uroarthritis (n = 37), or Reiter's syndrome, aetiology unknown (n = 5) were included in the study. Lumbosacral radiographs were taken 7 to 38 years after the initial arthritis. Three independent observers read the radiographs. Spinal changes included squaring of vertebrae, Romanus lesions, syndesmophytes, and osteophytes. Sacroiliitis was recorded according to the New York and Stoke methods. Signs of enthesitis in the iliac crest and disc space narrowing were recorded. Interobserver reliability and intraobserver reproducibility were determined. RESULTS: 23% of patients had grade 2-4 sacroiliitis (New York criteria) and 14% had syndesmophytes. There was more frequent sacroiliitis (32% v 13%) in uroarthritis than in enteroarthritis, and more syndesmophytes (mean 0.54 v 0.15 per patient; prevalence 24% v 6%, respectively). In other radiological features, no significant differences were present between the groups. In the material as a whole, patients with sacroiliitis of grade 2 or more had significantly less disc space narrowing both in patient comparisons (chi(2) test) and in numbers of spaces involved (Mann-Whitney test). Interobserver and intraobserver agreement, calculated using Cohen's kappa method, varied from 0.2 to 1. CONCLUSIONS: Syndesmophytes and sacroiliitis are more common in patients with previous uroarthritis than in those with previous enteroarthritis, but radiological findings in lumbosacral spine radiographs are characteristically similar. PMID- 15539416 TI - Arterial baroreflex alters strength and mechanisms of muscle metaboreflex during dynamic exercise. AB - Previous studies showed that the arterial baroreflex opposes the pressor response mediated by muscle metaboreflex activation during mild dynamic exercise. However, no studies have investigated the mechanisms contributing to metaboreflex-mediated pressor responses during dynamic exercise after arterial baroreceptor denervation. Therefore, we investigated the contribution of cardiac output (CO) and peripheral vasoconstriction in mediating the pressor response to graded reductions in hindlimb perfusion in conscious, chronically instrumented dogs before and after sinoaortic denervation (SAD) during mild and moderate exercise. In control experiments, the metaboreflex pressor responses were mediated via increases in CO. After SAD, the metaboreflex pressor responses were significantly greater and significantly smaller increases in CO occurred. During control experiments, nonischemic vascular conductance (NIVC) did not change with muscle metaboreflex activation, whereas after SAD NIVC significantly decreased with metaboreflex activation; thus SAD shifted the mechanisms of the muscle metaboreflex from mainly increases in CO to combined cardiac and peripheral vasoconstrictor responses. We conclude that the major mechanism by which the arterial baroreflex buffers the muscle metaboreflex is inhibition of metaboreflex mediated peripheral vasoconstriction. PMID- 15539415 TI - Analysis of the CD8+ T cell response to the G1 domain of aggrecan in ankylosing spondylitis. AB - BACKGROUND: CD4+ T cell responses to the G1 domain of aggrecan in patients with ankylosing spondylitis (AS) were recently reported. Whether such an immune response can be seen in the CD8+ subpopulation has not yet been determined. OBJECTIVE: To determine if HLA-B27 restricted G1-specific CD8+ T cells are present in AS and to analyse immunodominant CD8+ T cell epitopes. METHODS: Peripheral blood mononuclear cells of 45 patients with AS were stimulated with overlapping 18-mer peptides covering the whole G1 protein. Results were compared with those for patients with rheumatoid arthritis (RA) and healthy controls. For epitope analysis, G1-specific interferon gamma positive (IFNgamma+) T cells were isolated by magnetic activated cell sorting. After in vitro expansion, CD8+ T cells were restimulated with 14 subpools of G1 peptides. T cells responding to G1 peptide subpools were quantified by flow cytometry according to IFNgamma secretion. Predicted peptides were subsequently confirmed by stimulation with single peptides. RESULTS: G1-specific CD8+ T cell responses were found in 29/45 (64%) patients with AS, 18/35 (51%) patients with RA, but not in healthy controls. Five CD8+ T cell epitopes were identified as immunodominant in five patients. However, the T cell response was not HLA-B27 restricted. Nonamer peptides with an HLA-B27 binding motif did not induce a T cell response. CONCLUSION: A G1 peptide-specific CD8+ T cell response is present in AS but also in patients with RA. It does not seem to be HLA-B27 restricted. Whether such a response has a role in the pathogenesis of AS needs clarification. PMID- 15539417 TI - Acute adenosine preconditioning is mediated by p38 MAPK activation in discrete subcellular compartments. AB - Although acute adenosine preconditioning (PC) is well established, the signaling pathways mediating this cardioprotection remain unclear. Because adenosine receptor agonists activate p38 MAPK and this kinase has been implicated in ischemic and pharmacological PC, the purpose of this study was to determine the role of p38 MAPK in acute adenosine receptor PC. The role of p38 MAPK activation in discrete subcellular compartments during ischemia-reperfusion was also determined. The following groups were used in an in vivo rat ischemia-reperfusion model: 1) control (10% DMSO i.v.), 2) the A(1)/A(2a) adenosine receptor AMP-579 (50 microg/kg i.v.), 3) AMP-579 + the A(1) receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX, 100 microg/kg i.v.), 4) AMP-579 + the p38 MAPK inhibitor SB-203580 (1 mg/kg i.v.), and 5) SB-203580 alone. p38 MAPK activation was measured by Western blot analysis in cytosolic, mitochondrial, membrane, and nuclear/myofilament fractions obtained from hearts at preischemic, ischemic, and reperfusion time points. A significant reduction in infarct size was observed with AMP-579 PC, an effect blocked by DPCPX or SB-203580 pretreatment. AMP-579 treatment was associated with a significant increase in p38 MAPK activation in the nuclear/myofilament fraction before ischemia, whereas no activation of this kinase occurred during ischemia or reperfusion. In contrast, p38 MAPK was activated in the mitochondrial fraction by ischemia and in the cytosolic, mitochondrial, and membrane fractions by reperfusion in the control group. SB 203580 blocked the AMP-579-induced increase in phosphorylation of the downstream p38 substrate activating transcription factor-2. These results suggest a role for p38 MAPK activation in discrete subcellular compartments in acute adenosine A(1) receptor PC. PMID- 15539418 TI - Urinary trypsin inhibitor reduces LPS-induced hypotension by suppressing tumor necrosis factor-alpha production through inhibition of Egr-1 expression. AB - Although urinary trypsin inhibitor (UTI) has been shown to inhibit tumor necrosis factor (TNF)-alpha- production, the detailed mechanism(s) remains unclear. This study was undertaken to elucidate the molecular mechanism(s) underlying this inhibitory effect in monocytes in vitro and in rats given lipopolysaccharide (LPS). TNF-alpha production by monocytes stimulated with LPS (100 ng/ml) was inhibited by UTI at concentrations higher than 100 U/ml. Expression of early growth response factor-1 (Egr-1) and phosphorylation of extracellular signal regulated protein kinases 1/2 in monocytes stimulated with LPS were inhibited by UTI. UTI (50,000 U/kg i.v.) inhibited LPS (5 mg/kg i.v.)-induced increases in lung tissue levels of Egr-1, TNF-alpha mRNA, and TNF-alpha in rats. UTI inhibited LPS-induced hypotension by inhibiting pulmonary induction of inducible nitric oxide synthase (iNOS). We previously demonstrated that anti-TNF-alpha antibody and aminoguanidine, a selective inhibitor of iNOS, reduced LPS-induced hypotension in this animal model. Furthermore, we also reported that reduction of LPS-induced coagulation abnormalities in rats did not affect inflammatory responses and hypotension in this animal model. Taken together, these observations strongly suggested that UTI inhibited LPS-induced production of TNF alpha by inhibiting activation of the extracellular signal-regulated protein kinases 1/2-Egr-1 pathway in monocytes, which might at least partly contribute to reduction of hypotension through inhibition of iNOS induction in rats given LPS. PMID- 15539419 TI - Differential role of nitric oxide in regional sympathetic responses to stimulation of NTS A2a adenosine receptors. AB - Our previous studies showed that preganglionic adrenal (pre-ASNA), renal (RSNA), lumbar, and postganglionic adrenal sympathetic nerve activities (post-ASNA) are inhibited after stimulation of arterial baroreceptors, nucleus of the solitary tract (NTS), and glutamatergic and P2x receptors and are activated after stimulation of adenosine A1 receptors. However, stimulation of adenosine A2a receptors inhibited RSNA and post-ASNA, whereas it activated pre-ASNA. Because the effects evoked by NTS A2a receptors may be mediated via activation of nitric oxide (NO) mechanisms in NTS neurons, we tested the hypothesis that NO synthase (NOS) inhibitors would attenuate regional sympathetic responses to NTS A2a receptor stimulation, whereas NO donors would evoke contrasting responses from pre-ASNA versus RSNA and post-ASNA. Therefore, in chloralose/urethane anesthetized rats, we compared hemodynamic and regional sympathetic responses to microinjections of selective A2a receptor agonist (CGS-21680, 20 pmol/50 nl) after pretreatment with NOS inhibitors Nomega-nitro-L-arginine methyl ester (10 nmol/100 nl) and 1-[2-(trifluoromethyl)phenyl]imidazole (100 pmol/100 nl) versus pretreatment with vehicle (100 nl). In addition, responses to microinjections into the NTS of different NO donors [40 and 400 pmol/50 nl sodium nitroprusside (SNP); 0.5 and 5 nmol/50 nl 3,3-bis(aminoethyl)-1-hydroxy-2-oxo-1-triazene (DETA NONOate, also known as NOC-18), and 2 nmol/50 nl 3-(2-hydroxy-2-nitroso-1 propylhydrazino)-1-propanamine (PAPA NONOate, also known as NOC-15)], the NO precursor L-arginine (10-50 nmol/50 nl), and sodium glutamate (500 pmol/50 nl) were evaluated. SNP, DETA NONOate, and PAPA NONOate activated pre-ASNA and inhibited RSNA and post-ASNA, whereas l-arginine and glutamate microinjected into the same site of the NTS inhibited all these sympathetic outputs. Decreases in heart rate and depressor or biphasic responses accompanied the neural responses. Pretreatment with NOS inhibitors reversed the normal depressor and sympathoinhibitory responses to stimulation of NTS A2a receptors into pressor and sympathoactivatory responses and attenuated the heart rate decreases; however, it did not change the increases in pre-ASNA. We conclude that NTS NO mechanisms differentially affect regional sympathetic outputs and differentially contribute to the pattern of regional sympathetic responses evoked by stimulation of NTS A2a receptors. PMID- 15539420 TI - Mechanism of hypoalbuminemia in rodents. AB - Normal albumin loss from the plasma is thought to be minimized by a number of mechanisms, including charge repulsion with the capillary wall and an intracellular rescue pathway involving the major histocompatibility complex related Fc receptor (FcRn)-mediated mechanism. This study investigates how these factors may influence the mechanism of hypoalbuminemia. Hypoalbuminemia in rats was induced by treatment with puromycin aminonucleoside (PA). To test the effects of PA on capillary wall permeability, plasma elimination rates were determined for tritium-labeled tracers of different-sized Ficolls, negatively charged Ficolls, and (14)C-labeled tracer of albumin in control and PA-treated Sprague Dawley rats. Urinary excretion and tissue uptake were also measured. Hypoalbuminemia was also examined in two strains of FcRn-deficient mice: beta(2) microglobulin (beta(2)M) knockout (KO) mice and FcRn alpha-chain KO mice. The excretion rates of albumin and albumin-derived fragments were measured. PA induced hypoalbuminemia was associated with a 2.5-fold increase in the plasma elimination rate of albumin. This increase could be completely accounted for by the increase in urinary albumin excretion. Changes in the permeability of the capillary wall were not apparent, inasmuch as there was no comparable increase in the plasma elimination rate of 36- to 85-A Ficoll or negatively charged 50- to 80 A Ficoll. In contrast, hypoalbuminemic states in beta(2)M and FcRn KO mice were associated with decreases in excretion of albumin and albumin-derived fragments. This demonstrates that the mechanism of hypoalbuminemia consists of at least two distinct forms: one specifically associated with the renal handling of albumin and the other mediated by systemic processes. PMID- 15539421 TI - L-type Ca2+ channel function in the avian embryonic heart after cardiac neural crest ablation. AB - In avian and mammalian embryos, surgical ablation or severely reduced migration of the cardiac neural crest leads to a failure of outflow tract septation known as persistent truncus arteriosus (PTA) and leads to embryo lethality due partly to impaired excitation-contraction coupling stemming primarily from a reduction in the L-type Ca(2+) current (I(Ca),(L)). Decreased I(Ca,L) occurs without a corresponding reduction in the alpha(1)-subunit of the Ca(2+) channel. We hypothesize that decreased I(Ca),(L) is due to reduced function at the single channel level. The cell-attached patch clamp with Na(+) as the charge carrier was used to examine single Ca(2+) channel activity in myocytes from normal hearts from sham-operated embryos and from hearts diagnosed with PTA at embryonic days (ED) 11 and 15 after laser ablation of the cardiac neural crest. In normal hearts, the number of single channel events per 200-ms depolarization and the mean open channel probability (P(o)) was 1.89 +/- 0.17 and 0.067 +/- 0.008 for ED11 and 1.14 +/- 0.17 and 0.044 +/- 0.005 for ED15, respectively. These values represent a normal reduction in channel function and I(Ca),(L) observed with development. However, the number of single channel events was significantly reduced in hearts with PTA at both ED11 and ED15 (71% and 47%, respectively) with a corresponding reduction in P(o) (75% and 43%). The open time frequency histograms were best fitted by single exponentials with similar decay constants (tau approximately or equal 4.5 ms) except for the sham operated at ED15 (tau = 3.4 ms). These results indicate that the cardiac neural crest influences the development of myocardial Ca(2+) channels. PMID- 15539422 TI - Mathematical model of oxygen distribution in engineered cardiac tissue with parallel channel array perfused with culture medium containing oxygen carriers. AB - A steady-state model of oxygen distribution in a cardiac tissue construct with a parallel channel array was developed and solved for a set of parameters using the finite element method and commercial software (FEMLAB). The effects of an oxygen carrier [Oxygent; 32% volume perfluorocarbon (PFC) emulsion] were evaluated. The parallel channel array mimics the in vivo capillary tissue bed, and the PFC emulsion has a similar role as the natural oxygen carrier hemoglobin in increasing total oxygen content. The construct was divided into an array of cylindrical domains with a channel in the center and tissue space surrounding the channel. In the channel, the main modes of mass transfer were axial convection and radial diffusion. In the tissue region, mass transfer was by axial and radial diffusion, and the consumption of oxygen was by Michaelis-Menten kinetics. Neumann boundary conditions were imposed at the channel centerline and the half distance between the domains. Supplementation of culture medium by PFC emulsion improved mass transport by increasing convective term and effective diffusivity of culture medium. The model was first implemented for the following set of experimentally obtained parameters: construct thickness of 0.2 cm, channel diameter of 330 mum, channel center-to-center spacingof 700 microm, and average linear velocity per channel of 0.049 cm/s, in conjunction with PFC supplemented and unsupplemented culture medium. Subsequently, the model was used to define favorable scaffold geometry and flow conditions necessary to cultivate cardiac constructs of high cell density (10(8) cells/ml) and clinically relevant thickness (0.5 cm). In future work, the model can be utilized as a tool for optimization of scaffold geometry and flow conditions. PMID- 15539423 TI - Role of A1 adenosine receptors in regulation of vascular tone. AB - The vascular response to adenosine and its analogs is mediated by four adenosine receptors (ARs), namely, A(1), A(2A), A(2B), and A(3). A(2A)ARs and/or A(2B)ARs are involved in adenosine-mediated vascular relaxation of coronary and aortic beds. However, the role of A(1)ARs in the regulation of vascular tone is less well substantiated. The aim of this study was to determine the role of A(1)ARs in adenosine-mediated regulation of vascular tone. A(1)AR-knockout [A(1)AR((-/-))] mice and available pharmacological tools were used to elucidate the function of A(1)ARs and the impact of these receptors on the regulation of vascular tone. Isolated aortic rings from A(1)AR((-/-)) and wild-type [A(1)AR((+/+))] mice were precontracted with phenylephrine, and concentration-response curves for adenosine and its analogs, 5'-N-ethyl-carboxamidoadenosine (NECA, nonselective), 2-chloro N(6)-cyclopentyladenosine (CCPA, A(1)AR selective), 2-(2-carboxyethyl)phenethyl amino-5'-N-ethylcarboxamido-adenosine (CGS-21680, A(2A) selective), and 2-chloro N(6)-3-iodobenzyladenosine-5'-N-methyluronamide (Cl-IBMECA, A(3) selective) were obtained to determine relaxation. Adenosine and NECA (0.1 microM) caused small contractions of 13.9 +/- 3.0 and 16.4 +/- 6.4%, respectively, and CCPA at 0.1 and 1.0 microM caused contractions of 30.8 +/- 4.3 and 28.1 +/- 3.9%, respectively, in A(1)AR((+/+)) rings. NECA- and CCPA-induced contractions were eliminated by 100 nM of 1,3-dipropyl-8-cyclopentylxanthine (DPCPX, selective A(1)AR antagonist). Adenosine, NECA, and CGS-21680 produced an increase in maximal relaxation in A(1)AR((-/-)) compared with A(1)AR((+/+)) rings, whereas Cl-IBMECA did not produce contraction in either A(1)AR((+/+)) or A(1)AR((-/-)) rings. CCPA induced contraction at 1.0 microM was eliminated by the PLC inhibitor U-73122. These data suggest that activation of A(1)ARs causes contraction of vascular smooth muscle through PLC pathways and negatively modulates the vascular relaxation mediated by other adenosine receptor subtypes. PMID- 15539424 TI - Effects of pre-, peri-, and postmyocardial infarction treatment with losartan in rats: effect of dose on survival, ventricular arrhythmias, function, and remodeling. AB - Angiotensin receptor blockers (ARBs) reduce adverse left ventricular (LV) remodeling and improve LV function and survival when started postmyocardial infarction (MI). ARBs also reduce ventricular arrhythmias during ischemia reperfusion injury when started pre-MI. No information exists regarding their efficacy and safety when started pre-MI and continued peri- and post-MI. We evaluated whether the ARB losartan improves the outcome when started pre-MI and continued peri- and post-MI. Male Wistar rats (n = 502) were treated for 7 days pre-MI with losartan at a high dose (30 mg.kg(-1).day(-1)), progressively increasing dose (3 mg.kg(-1).day(-1) increased to 10 mg.kg(-1).day(-1) 10 days and 30 mg.kg(-1).day(-1) 20 days post-MI), or no treatment. Ambulatory systolic blood pressure and Holter monitoring were performed for 24 h post-MI. Echocardiography was done 30 days post-MI, and LV remodeling, cardiac hemodynamics, and fetal gene expression were assessed 38 days post-MI. High-dose losartan reduced 24-h post-MI survival compared with the progressive dose and control (21.9% vs. 36.6% and 38.1%, P = 0.033 and P = 0.009, respectively). This was associated with greater hypotension in the high dose and no change in ventricular arrhythmias in all groups. In 24-h post-MI survivors, the progressive dose group had reduced mortality from 24 h to 38 days (8.5% vs. 28.6% for control vs. 38.9% for high dose, P = 0.032 and P = 0.01, respectively). Survivors of both losartan groups demonstrated improved LV remodeling, cardiac hemodynamics, preserved GLUT-4, and reduced cardiac fetal gene expression. Pretreatment with ARBs does not reduce 24-h post-MI ventricular arrhythmias or survival, and high doses increase mortality by causing excessive hypotension. In 24-h post-MI survivors, progressively increasing doses of losartan have multiple beneficial effects, including improved survival. PMID- 15539425 TI - Contribution of KATP+ channels to coronary vasomotor tone regulation is enhanced in exercising swine with a recent myocardial infarction. AB - Previous studies demonstrated a decreased flow reserve in the hypertrophied myocardium early after myocardial infarction (MI). Previously, we reported that exacerbation of hemodynamic abnormalities and neurohumoral activation during exercise caused slight impairment of myocardial O(2) supply in swine with a recent MI. We hypothesized that increased metabolic coronary vasodilation [via ATP-sensitive K(+) (K(ATP)(+)) channels and adenosine] may have partially compensated for the increased extravascular compressive forces and increased vasoconstrictor neurohormones, thereby preventing a more severe impairment of myocardial O(2) balance. Chronically instrumented swine were exercised on a treadmill up to 85% of maximum heart rate. Under resting conditions, adenosine receptor blockade [8-phenyltheophylline (8-PT), 5 mg/kg i.v.] and K(ATP)(+) channel blockade (glibenclamide, 3 mg/kg i.v.) produced similar decreases in myocardial O(2) supply in normal and MI swine. However, while glibenclamide's effect waned in normal swine during exercise (P < 0.05), it was maintained in MI swine. 8-PT's effect was maintained during exercise and was not different between normal and MI swine. Finally, in normal swine combined treatment with 8-PT and glibenclamide produced a vasoconstrictor response that equaled the sum of the responses to blockade of the individual pathways. In contrast, in MI swine the vasoconstrictor response to 8-PT and glibenclamide was similar to that produced by glibenclamide alone. In conclusion, despite significant hemodynamic abnormalities in swine with a recent MI, myocardial O(2) supply and O(2) consumption in remodeled myocardium are still closely matched during exercise. This close matching is supported by increased K(ATP)(+) channel-mediated coronary vasodilation. Although the net vasodilator influence of adenosine was unchanged in remodeled myocardium, it became exclusively dependent on K(ATP)(+) channel opening. PMID- 15539426 TI - Redox regulation of Ito remodeling in diabetic rat heart. AB - Oxidative stress and the resulting change in cell redox state are proposed to contribute to pathogenic alterations in ion channels that underlie electrical remodeling of the diseased heart. The present study examined whether K(+) channel remodeling is controlled by endogenous oxidoreductase systems that regulate redox sensitive cell functions. Diabetes was induced in rats by streptozotocin, and experiments were conducted after 3-5 wk of hyperglycemia. Spectrophotometric assays of ventricular tissue extracts from diabetic rat hearts revealed divergent changes in two major oxidoreductase systems. The thioredoxin (TRX) system in diabetic rat heart was characterized by a 52% decrease in TRX reductase (TRXR) activity from control heart (P < 0.05), whereas TRX activity was 1.7-fold greater than control heart (P < 0.05). Diabetes elicited similar changes in the glutaredoxin (GRX) system: glutathione reductase was decreased 35% from control level (P < 0.05), and GRX activity was 2.5-fold greater than in control heart (P < 0.05). The basal activity of glucose-6-phosphate dehydrogenase, which generates NADPH required by the TRX and GRX systems, was not altered by diabetes. Voltage clamp studies showed that the characteristically decreased density of the transient outward K(+) current (I(to)) in isolated diabetic rat myocytes was normalized by in vitro treatment with insulin (0.1 microM) or the metabolic activator dichloroacetate (1.5 mM). The effect of these agonists on I(to) was blocked by inhibitors of glucose-6-phosphate dehydrogenase. Moreover, inhibitors of TRXR, which controls the reducing activity of TRX, also blocked upregulation of I(to) by insulin and dichloroacetate. These data suggest that K(+) channels underlying I(to) are regulated in a redox-sensitive manner by the TRX system and the remodeling of I(to) that occurs in diabetes may be due to decreased TRXR activity. We propose that oxidoreductase systems are an important repair mechanism that protects ion channels and associated regulatory proteins from irreversible oxidative damage. PMID- 15539427 TI - Adrenomedullin enhances therapeutic potency of bone marrow transplantation for myocardial infarction in rats. AB - Adrenomedullin (AM), a potent vasodilator, induces angiogenesis and inhibits cell apoptosis through the phosphatidylinositol 3-kinase/Akt pathway. Transplantation of bone marrow-derived mononuclear cells (MNC) induces angiogenesis. We investigated whether infusion of AM enhances the therapeutic potency of MNC transplantation in a rat model of myocardial infarction. Immediately after coronary ligation, bone marrow-derived MNC (5 x 10(6) cells) were injected into the ischemic myocardium, followed by subcutaneous administration of 0.05 microg x kg(-1) x min(-1) AM (AM-MNC group) or saline (MNC group) for 3 days. Another two groups of rats received subcutaneous administration of AM alone (AM group) or saline (control group). Hemodynamic and histological analyses were performed 4 wk after treatment. Cardiac infarct size was significantly smaller in the MNC and AM groups than in the control group. A combination of AM infusion and MNC transplantation demonstrated a further decrease in infarct size. Left ventricular (LV) maximum change in pressure over time and LV fractional shortening were significantly improved only in the AM-MNC group. AM significantly increased capillary density in ischemic myocardium, suggesting the angiogenic potency of AM. AM infusion plus MNC transplantation demonstrated a further increase in capillary density compared with AM or MNC alone. Although MNC apoptosis was frequently observed 72 h after transplantation, AM markedly decreased the number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling-positive cells among the transplanted MNC. In conclusion, AM enhanced the angiogenic potency of MNC transplantation and improved cardiac function in rats with myocardial infarction. This beneficial effect may be mediated partly by the angiogenic property of AM itself and by its antiapoptotic effect on MNC. PMID- 15539428 TI - Glucocorticoid receptor ligand binding domain is sufficient for the modulation of glucocorticoid induction properties by homologous receptors, coactivator transcription intermediary factor 2, and Ubc9. AB - Several factors modulate the position of the dose-response curve of steroid receptor-agonist complexes and the partial agonist activity of antagonist complexes, thereby causing differential gene activation by circulating hormones and unequal gene repression during endocrine therapies with antisteroids. We now ask whether the modulatory activity of three factors (homologous receptor, coactivator transcription intermediary factor 2, and Ubc9) requires the same or different domains of glucocorticoid receptors (GRs). In all cases, we find that neither the amino terminal half of the receptor, which contains the activation function-1 activation domain, nor the DNA binding domain is required. This contrasts with the major role of activation function-1 in determining the amount of gene expression and partial agonist activity of antisteroids with most steroid receptors. However, the situation is more complicated with Ubc9, where GR N terminal sequences prevent the actions of Ubc9, but not added GR or transcription intermediary factor 2, at low GR concentrations. Inhibition is relieved by deletion of these sequences or by replacement with the comparable region of progesterone receptors but not by overexpression of the repressive sequences. These results plus the binding of C-terminal GR sequences to the suppressive N terminal domain implicate an intramolecular mechanism for the inhibition of Ubc9 actions at low GR concentrations. A shift from noncooperative to cooperative steroid binding at high GR concentrations suggests that conformational changes reposition the inhibitory N-terminal sequence to allow Ubc9 interaction with elements of the ligand binding domain. Collectively, these results indicate a dominant role of GR C-terminal sequences in the modulation of the dose-response curve and partial agonist activity of GR complexes. They also reveal mechanistic differences both among individual modulators and between the ability of the same factors to regulate the total amount of gene expression. PMID- 15539429 TI - Evidence that palmitoylation of carboxyl terminus cysteine residues of the human luteinizing hormone receptor regulates postendocytic processing. AB - Palmitoylation is a well-conserved posttranslational modification among members of the G protein-coupled receptor superfamily. The present study examined the role of palmitoylation in endocytosis and postendocytic trafficking of the human LH receptor (LHR). Palmitoylation of the LHR was determined by incorporation of [3H]palmitic acid into wild-type (WT) or mutant receptor in which the potential palmitoylation sites, C643 and C644, were mutated to glycine residues. The WT receptor showed incorporation of [3H]palmitic acid into the mature 90-kDa form of the receptor whereas mutation of the two Cys residues abrogated this incorporation, indicating that Cys 643 and C644 are the sites of palmitoylation. The role of palmitoylation on endocytosis and postendocytic processing was examined by testing the ability of the WT and mutant receptor to undergo internalization, recycling, and lysosomal degradation. Compared with the WT receptor, the mutant receptor showed increased internalization and decreased recycling, suggesting that retention of palmitic acid residues at Cys 643 and 644 promotes LHR recycling. The role of palmitoylation on receptor recycling was substantiated by demonstrating that a different mutant, D578H LHR, which is deficient in palmitoylation, also recycled less efficiently. Furthermore, the data show that palmitoylation, not the rate of internalization, determines the efficiency of recycling. The present study shows that palmitoylation of cysteine residues 643 and 644 of the human LHR is a determinant of recycling. PMID- 15539430 TI - Cyclin D1 binding to the androgen receptor (AR) NH2-terminal domain inhibits activation function 2 association and reveals dual roles for AR corepression. AB - The androgen receptor (AR) is a member of the nuclear receptor superfamily, the activity of which is critical for the development and progression of prostate cancer. We and others have previously demonstrated that cyclin D1 is a potent corepressor of the AR. Although cyclin D1 is suspected to recruit histone deacetylases to the AR complex, previous studies have demonstrated that this activity alone is insufficient for cyclin D1 function. Here, we uncover a novel, secondary means of cyclin D1-mediated repression, through modulation of AR amino carboxy terminal interactions. We show that cyclin D1 predominantly binds the N terminal domain of the AR, dependent on the AR 23FxxLF27 motif. Through this motif, cyclin D1 abrogates the ability of the AR N-terminal domain to interact with the C terminus. Secondary amino-terminal domain sites capable of fostering interaction with the C terminus were refractory to cyclin D1 action, indicating that the ability of cyclin D1 to modulate AR amino-carboxy terminal interactions is specific to 23FxxLF27. Deletion of the N-terminal cyclin D1 binding site severely compromised AR activity (due to loss of FxxLF) but unmasked a repressor action through interaction with the AR C terminus. In summary, these data reveal novel, unexpected mechanisms of cyclin D1 activity and demonstrate that this function of cyclin D1 is critical for AR modulation. PMID- 15539431 TI - The zinc finger-containing transcription factor Gata-4 is expressed in the developing endocrine pancreas and activates glucagon gene expression. AB - Gene inactivation studies have shown that members of the Gata family of transcription factors are critical for endoderm development throughout evolution. We show here that Gata-4 and/or Gata-6 are not only expressed in the adult exocrine pancreas but also in glucagonoma and insulinoma cell lines, whereas Gata 5 is restricted to the exocrine pancreas. During pancreas development, Gata-4 is expressed already at embryonic d 10.5 and colocalizes with early glucagon+ cells at embryonic d 12.5. Gata-4 was able to transactivate the glucagon gene both in heterologous BHK-21 (nonislet Syrian baby hamster kidney) and in glucagon producing InR1G9 cells. Using gel-mobility shift assays, we identified a complex formed with nuclear extracts from InR1G9 cells on the G5 control element (-140 to -169) of the glucagon gene promoter as Gata-4. Mutation of the GATA binding site on G5 abrogated the transcriptional activation mediated by Gata-4 and reduced basal glucagon gene promoter activity in glucagon-producing cells by 55%. Furthermore, Gata-4 acted more than additively with Forkhead box A (hepatic nuclear factor-3) to trans-activate the glucagon gene promoter. We conclude that, besides its role in endoderm differentiation, Gata-4 might be implicated in the regulation of glucagon gene expression in the fetal pancreas and that Gata activity itself may be modulated by interactions with different cofactors. PMID- 15539432 TI - Novel and quantitative DNA dot-blotting method for assessment of in vivo proliferation. AB - Immunohistochemical assessment of 5-bromo-2-deoxyuridine (BrdU) in tissue sections is a widely used method to evaluate cell proliferation in vivo. However, this method requires time-consuming preparation of paraffin sections and laborious counting of BrdU-labeled nuclei on multiple sections. Here, we report the development of a rapid and reliable method to quantitate BrdU incorporation in intestinal and liver tissues using a dot-blot method. In vivo models of colon or liver proliferation were used to analyze the usefulness and reliability of this new method. Mice were killed after BrdU injection, and genomic DNA was isolated from the tissues, denatured, and dot-blotted onto a nitrocellulose membrane. The incorporated BrdU was detected with a BrdU monoclonal antibody, and the signal intensity was densitometrically quantified. Results were compared with BrdU index determined by conventional immunohistochemistry on the same tissue samples. The patterns of colonic BrdU incorporation during fasting and refeeding, measured by the dot-blotting method and the immunohistochemical method, were similar. The BrdU incorporation in the regenerating liver after partial hepatectomy, evaluated by these two different methods, showed a strong correlation (R(2) = 0.91, P < 0.01). In addition, the inhibition of colon proliferation by the phosphoinositol 3-kinase inhibitor wortmannin was demonstrated by this dot-blotting method. In conclusion, the dot-blotting technique described in this report provides an accurate, more efficient, and possibly more reliable method for the assessment of in vivo proliferation compared with conventional immunohistochemical determination of BrdU-labeling index. PMID- 15539433 TI - Effects of Boswellia serrata in mouse models of chemically induced colitis. AB - Extracts from Boswellia serrata have been reported to have anti-inflammatory activity, primarily via boswellic acid-mediated inhibition of leukotriene synthesis. In three small clinical trials, boswellia was shown to improve symptoms of ulcerative colitis and Crohn's disease, and because of its alleged safety, boswellia was considered superior over mesalazine in terms of a benefit risk evaluation. The goal of this study was to evaluate the effectiveness of boswellia extracts in controlled settings of dextran sulfate- or trinitrobenzene sulfonic acid-induced colitis in mice. Our results suggest that boswellia is ineffective in ameliorating colitis in these models. Moreover, individual boswellic acids were demonstrated to increase the basal and IL-1beta-stimulated NF-kappaB activity in intestinal epithelial cells in vitro as well as reverse proliferative effects of IL-1beta. We also observed hepatotoxic effect of boswellia with pronounced hepatomegaly and steatosis. Hepatotoxity and increased lipid accumulation in response to boswellia were further confirmed in vitro in HepG2 cells with fluorescent Nile red binding/resazurin reduction assay and by confocal microscopy. Microarray analyses of hepatic gene expression demonstrated dysregulation of a number of genes, including a large group of lipid metabolism related genes, and detoxifying enzymes, a response consistent with that to hepatotoxic xenobiotics. In summary, boswellia does not ameliorate symptoms of colitis in chemically induced murine models and, in higher doses, may become hepatotoxic. Potential implications of prolonged and uncontrolled intake of boswellia as an herbal supplement in inflammatory bowel disease and other inflammatory conditions should be considered in future clinical trials with this botanical. PMID- 15539434 TI - Muscarinic activation of Na+-dependent ion transporters and modulation by bicarbonate in rat submandibular gland acinus. AB - To investigate the interaction between the ion channels and transporters in the salivary fluid secretion, we measured the membrane voltage (V(m)) and intracellular concentrations of Ca(2+), Na(+) ([Na(+)](c)), Cl(-), and H(+) (pH(i)) in rat submandibular gland acini (RSMGA). After a transient depolarization induced by a short application of acetylcholine (ACh; 5 muM, 20 s), RSMGA showed strong delayed hyperpolarization (V(h,ACh); -95 +/- 1.8 mV) that was abolished by ouabain. In the HCO(3)(-)-free condition, the V(h,ACh) was also blocked by bumetanide, a blocker of Na(+)-K(+)-2Cl(-) cotransporter (NKCC). In the presence of HCO(3)(-) (24 meq, bubbled with 5% CO(2)), however, the V(h,ACh) was not blocked by bumetanide, but it was suppressed by ethylisopropylamiloride (EIPA), a Na(+)/H(+) exchanger (NHE) inhibitor. Similarly, the ACh-induced increase in [Na(+)](c) was totally blocked by bumetanide in the absence of HCO(3)(-), but only by one-half in the presence of HCO(3)(-). ACh induced a prominent acidification of pH(i) in the presence of HCO(3)(-), and the acidification was further increased by EIPA treatment. Without HCO(3)(-), an application of ACh strongly accelerated the NKCC activity that was measured from the decay of pH(i) during the application of NH(4)(+) (20 mM). Notably, the ACh induced activation of NKCC was largely suppressed in the presence of HCO(3)(-). In summary, the ACh-induced anion secretion in RSMGA is followed by the activation of NKCC and NHE, resulting an increase in [Na(+)](c). The intracellular Na(+)-induced activation of electrogenic Na(+)/K(+)-ATPase causes V(h,ACh). The regulation of NKCC and NHE by ACh is strongly affected by the physiological level of HCO(3)(-). PMID- 15539435 TI - Association between sex hormone-binding globulin levels and activated protein C resistance in explaining the risk of thrombosis in users of oral contraceptives containing different progestogens. AB - BACKGROUND: Epidemiological studies have shown that both the estrogen dose and progestogen type of oral contraceptives contribute to the increased risk of thrombosis in oral contraceptive users. Thrombin generation-based activated protein C (APC) sensitivity is a global test for the net prothrombotic effect of oral contraceptives and predicts the thrombotic risk. Our objective was to test the usefulness of sex hormone-binding globulin (SHBG) as a marker for the thrombotic risk of an oral contraceptive. METHODS: We measured SHBG and APC resistance in 156 healthy users of various types of oral contraceptives. RESULTS: Users of oral contraceptives with a moderately increased risk of thrombosis (gestodene and desogestrel pills) had higher SHBG levels than users of low-risk oral contraceptives containing levonorgestrel. Similarly, for higher doses of estrogen in oral contraceptives we found higher SHBG levels. Women using oral contraceptives with the highest thrombotic risk (cyproterone acetate pills) rendered the highest SHBG levels. Users of oral contraceptives containing gestodene, desogestrel or cyproterone acetate were more resistant to APC than users of levonorgestrel pills. SHBG levels were positively associated with the increased APC resistance. CONCLUSIONS: Our findings support the hypothesis that the effect of an oral contraceptive on SHBG levels might be a marker for the thrombotic risk. PMID- 15539436 TI - Impaired insulin-dependent glucose metabolism in granulosa-lutein cells from anovulatory women with polycystic ovaries. AB - BACKGROUND: Insulin resistance and hyperinsulinaemia are well-recognized characteristics of anovulatory women with polycystic ovary syndrome (PCOS) but, paradoxically, steroidogenesis by PCOS granulosa cells remains responsive to insulin. The hypothesis to be tested in this study is that insulin resistance in the ovary is confined to the metabolic effects of insulin (i.e. glucose uptake and metabolism), whereas the steroidogenic action of insulin remains intact. METHODS: Granulosa-lutein cells were obtained during IVF cycles from seven women with normal ovaries, six ovulatory women with PCO (ovPCO) and seven anovulatory women with PCO (anovPCO). Mean body mass index was in the normal range in all three groups. Granulosa-lutein cells were cultured with insulin (1, 10, 100 and 1000 ng/ml) and LH (1, 2.5 and 5 ng/ml). Media were sampled at 24 and 48 h and analysed for glucose uptake, lactate production and (48 h only) progesterone production. RESULTS: Insulin-stimulated glucose uptake by cells from anovPCO was attenuated at higher doses of insulin (100 and 1000 ng/ml) compared with that by cells from either ovPCO (P=0.02) or controls (P=0.02). Insulin and LH stimulated lactate production in a dose-dependent manner, but insulin-dependent lactate production was markedly impaired in granulosa-lutein cells from anovPCO compared with either normal (P=0.002) or ovPCO (P<0.0001). By contrast, there was no difference in insulin-stimulated progesterone production between granulosa-lutein cells from the three ovarian types. CONCLUSIONS: Granulosa-lutein cells from women with anovPCOS are relatively resistant to the effects of insulin-stimulated glucose uptake and utilization compared with those from normal and ovPCO, whilst maintaining normal steroidogenic output in response to physiological doses of insulin. These studies support the probability of a post-receptor, signalling pathway-specific impairment of insulin action in PCOS. PMID- 15539437 TI - Cost-effectiveness of Chlamydia antibody tests in subfertile women. AB - BACKGROUND: For the evaluation of tubal function, Chlamydia antibody testing (CAT) has been introduced as a screening test. We compared six CAT screening strategies (five CAT tests and one combination of tests), with respect to their cost-effectiveness, by using IVF pregnancy rate as outcome measure. METHODS: A decision analytic model was developed based on a source population of 1715 subfertile women. The model incorporates hysterosalpingography (HSG), laparoscopy and IVF. To calculate IVF pregnancy rates, costs, effects, cost-effectiveness and incremental costs per effect of the six different CAT screening strategies were determined. RESULTS: pELISA Medac turned out to be the most cost-effective CAT screening strategy (15 075 per IVF pregnancy), followed by MIF Anilabsystems (15 108). A combination of tests (pELISA Medac and MIF Anilabsystems; 15 127) did not improve the cost-effectiveness of the single strategies. Sensitivity analyses showed that the results are robust for changes in the baseline values of the model parameters. CONCLUSIONS: Only small differences were found between the screening strategies regarding the cost-effectiveness, although pELISA Medac was the most cost-effective strategy. Before introducing a particular CAT test into clinical practice, one should consider the effects and consequences of the entire screening strategy, instead of only the diagnostic accuracy of the test used. PMID- 15539439 TI - No association of estrogen receptor alpha and cytochrome P450c17alpha polymorphisms with age at menopause in a Dutch cohort. AB - BACKGROUND: Age at menopause is under strong genetic control. So far, genetic variations of only one gene, the PvuII polymorphism of the estrogen receptor alpha (ERalpha) gene, have been shown to be associated with age at onset of menopause. This study aims to investigate whether PvuII, XbaI and B-variant polymorphisms of the ERalpha gene, and the MspAI polymorphism of the cytochrome P450c17alpha (CYP17) gene are associated with age at menopause in a Dutch cohort. METHODS: DNA was isolated from urine samples of 385 Caucasian women with natural menopause and the genotypes of the four polymorphisms were determined. A questionnaire was used for background characteristics. The genotypes of PvuII, XbaI, MspAI were obtained by PCR restriction fragment length polymorphism analysis. The B-variant was determined with an allele-specific oligonucleotide hybridization method. Two-sided t-tests were performed to assess the association between the four polymorphisms and menopausal age. The PvuII and XbaI polymorphisms were analysed separately as well as in a combined score. RESULTS: The results show that none of the polymorphisms independently, nor the combined genotypes for PvuII and XbaI, were associated with age at natural menopause. CONCLUSION: No evidence was found for a relationship between common variants of the ERalpha gene and the CYP17 gene with age at natural menopause. PMID- 15539438 TI - Superior cycle control with a contraceptive vaginal ring compared with an oral contraceptive containing 30 microg ethinylestradiol and 150 microg levonorgestrel: a randomized trial. AB - BACKGROUND: This trial was conducted to compare cycle control with vaginal ring a combined contraceptive vaginal ring, and a combined oral contraceptive (COC) delivering 30 mug ethinylestradiol (EE) and 150 mug levonorgestrel. METHODS: This open-label, randomized, multi-centre, Phase III study involved adult women from 11 countries. Subjects were treated with either vaginal ring or a COC for 13 cycles (12 months). RESULTS: A total of 1030 subjects (vaginal ring, n=512; COC, n=518) comprised the intention-to-treat (ITT) population. The percentage of women in the ITT population who completed the trial was 70.9% for vaginal ring and 71.2% for the COC group. The incidence of breakthrough bleeding and spotting over cycles 2-13, the primary efficacy parameter, was lower with vaginal ring (range 2.0-6.4%) than the COC (range 3.5-12.6%), and for cycles 2 and 9 the lower incidence with vaginal ring was confirmed as statistically significant (P=0.003 and P=0.002 respectively). The incidence of intended bleeding was significantly higher over all cycles with vaginal ring (58.8-72.8%) than with the COC (43.4 57.9%). CONCLUSIONS: Cycle control with vaginal ring was excellent and superior to that of a COC containing 30 mug EE. PMID- 15539440 TI - The pain-relieving effect of electro-acupuncture and conventional medical analgesic methods during oocyte retrieval: a systematic review of randomized controlled trials. AB - OBJECTIVE: The primary objective of the present review was to determine what pain relieving effect had been reported for acupuncture and other conscious sedation methods in assisted reproduction therapy since 1990. The secondary objective was to determine pregnancy rates, when possible. METHODS: The data source was the Medline database of the National Library of Medicine covering the period January 1990-January 2004. Bibliographies of relevant publications and review articles were scanned. A systematic review and meta-analyses of randomized, controlled trials comparing the pain-relieving effect of acupuncture and other conscious sedation methods was carried out. RESULTS: Of the 30 trials identified, 12 met the selection criteria for this systematic review and were included in the analysis. Five of the 12 studies reported differences in pain experiences during oocyte aspiration, but it was only possible to group the three trials evaluating the effect of electro-acupuncture (EA). The outcomes of these three studies were homogenous except from maximal and average pain. CONCLUSION: No method could be regarded as being superior to another, and no consensus on which method is optimal for pain relief during oocyte retrieval was found. Low doses of lignocaine can, however, be recommended in paracervical block (PCB) as well as EA without pre-medication. The clinical pregnancy rates appeared to be similar between the studies. Since pain experience varies from individual to individual, the optimal method of conscious sedation may also be individualized. PMID- 15539441 TI - Efficient treatment of infertility due to sperm DNA damage by ICSI with testicular spermatozoa. AB - BACKGROUND: Sperm DNA damage (fragmentation) is a recently discovered cause of male infertility for which no efficient treatment has yet been found. Previous findings have suggested that clinically relevant sperm DNA damage may occur at the post-testicular level. This study was undertaken to assess the clinical usefulness of ICSI with testicular spermatozoa in this indication. METHODS: The percentage of spermatozoa with fragmented DNA, assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling assay, and ICSI outcomes were compared in two sequential attempts performed, respectively, with ejaculated and testicular spermatozoa in 18 men with increased sperm DNA fragmentation. RESULTS: The incidence of DNA fragmentation was markedly lower in testicular spermatozoa as compared with ejaculated spermatozoa. No differences in fertilization and cleavage rates and in embryo morphological grade were found between the ICSI attempts performed with ejaculated and with testicular spermatozoa. However, eight ongoing clinical pregnancies (four singleton and four twin) were achieved by ICSI with testicular spermatozoa (44.4% pregnancy rate; 20.7% implantation rate), whereas ICSI with ejaculated spermatozoa led to only one pregnancy which was spontaneously aborted. CONCLUSIONS: These data show that ICSI with testicular spermatozoa provides the first efficient assisted reproduction treatment option for men with high levels of sperm DNA damage. PMID- 15539442 TI - Families created by gamete donation: follow-up at age 2. AB - BACKGROUND: Findings are presented of the second phase of a longitudinal study of families created through gamete donation. METHODS: At the time of the child's second birthday, 46 donor insemination families and 48 egg donation families were compared with 68 natural conception families on standardized interview and questionnaire measures of the psychological well being of the parents, parent child relationships and the psychological development of the child. RESULTS: The gamete donation mothers showed a trend towards greater pleasure in their child accompanied by a perception of their child as more vulnerable, with egg donation mothers tending towards greater pleasure and donor insemination mothers tending towards greater concern. The fathers did not differ on any of the variables under study. CONCLUSIONS: The findings from this recent cohort add further weight to the growing body of research showing that the absence of a genetic link between a parent and a child does not necessarily jeopardize the development of a positive relationship between them. PMID- 15539443 TI - Adolescents with open-identity sperm donors: reports from 12-17 year olds. AB - BACKGROUND: Donor insemination programs can include 'open-identity' sperm donors, who are willing to release their identities to adult offspring. We report findings from adolescent offspring who have open-identity donors. METHODS: Using mail-back questionnaires, youths from 29 households (41.4% headed by lesbian couples, 37.9% by single women, 20.7% by heterosexual couples) reported their experience growing up knowing how they were conceived and their interest in the donor's identity. RESULTS: Most youths (75.9%) reported always knowing, and were somewhat to very comfortable with their conception origins. All but one felt knowing had a neutral to positive impact on their relationship with their birth mother and, separately, co-parent. The youths' top question about the donor was, 'What's he like?' and >80% felt at least moderately likely to request his identity and pursue contact. Finally, of those who might contact the donor, 82.8% would do so to learn more about him, with many believing it would help them learn more about themselves. No youth reported wanting money and few (6.9%) wanted a father/child relationship. We also discuss differences found among youths from different household types. CONCLUSIONS: The majority of the youths felt comfortable with their origins and planned to obtain their donor's identity, although not necessarily at age 18. PMID- 15539444 TI - ESHRE PGD Consortium 'Best practice guidelines for clinical preimplantation genetic diagnosis (PGD) and preimplantation genetic screening (PGS)'. AB - Among the many educational materials produced by the European Society of Human Reproduction and Embryology (ESHRE) are guidelines. ESHRE guidelines may be developed for many reasons but their intent is always to promote best quality practices in reproductive medicine. In an era in which preimplantation genetic diagnosis (PGD) has become a reality, we must strive to maintain its efficacy and credibility by offering the safest and most effective treatment available. The dominant motivators for the development of current comprehensive guidelines for best PGD practice were (i) the absence of guidelines and/or regulation for PGD in many countries and (ii) the observation that no consensus exists on many of the clinical and technical aspects of PGD. As a consequence, the ESHRE PGD Consortium undertook to draw up guidelines aimed at giving information, support and guidance to potential, fledgling and established PGD centres. The success of a PGD treatment cycle is the result of great attention to detail. We have strived to provide a similar level of detail in this document and hope that it will assist staff in achieving the best clinical outcome for their patients. PMID- 15539445 TI - Molecular markers for the assessment of postnatal male germ cell development in the mouse. AB - BACKGROUND: A proliferation marker, proliferating cell nuclear antigen (PCNA), a Sertoli cell specific transcription factor, GATA-1 and the male germ cell specific, RNA binding motif (RBM), were used to identify different cellular populations during postnatal development of the mouse testis. METHODS: Immunohistochemistry, RT-PCR and real-time quantitative RT-PCR (QRT-PCR) were used. RESULTS: PCNA was expressed in pre-Sertoli and germ cells on the day of birth. Both pre-meiotic germ cells and spermatocytes expressed RBM throughout postnatal development. RBM-positive cell counts and QRT-PCR of RBM showed that average level of RBM per cell is highest in juvenile males between 14 and 21 days. From 42 days onward, there was a dramatic decrease in RBM expression in individual pre-meiotic and meiotic germ cells. CONCLUSIONS: These markers were used to correlate cell proliferative capability, gene expression profile and anatomic location within the developing mouse testis. The majority of germ cells start active proliferation once they have migrated to the basement membrane or immediately before. RBM is more highly expressed during the first wave of spermatogenesis versus subsequent waves, suggesting that there may be a change in the activity of RBM. PMID- 15539446 TI - A simple tool for drawing proteolytic peptide maps. AB - I describe a simple standalone program that assists in the preparation of peptide digestion maps. These are useful for comparative studies and for locating peptides within a primary sequence. The program creates an output file as scalable vector graphics that can then be viewed in a web browser or imported into a graphics-editing program. AVAILABILITY: The program, as a standalone executable, is available upon request from the author. CONTACT: r.beynon@liv.ac.uk. PMID- 15539447 TI - Predicting fold novelty based on ProtoNet hierarchical classification. AB - MOTIVATION: Structural genomics projects aim to solve a large number of protein structures with the ultimate objective of representing the entire protein space. The computational challenge is to identify and prioritize a small set of proteins with new, currently unknown, superfamilies or folds. RESULTS: We develop a method that assigns each protein a likelihood of it belonging to a new, yet undetermined, structural superfamily. The method relies on a variant of ProtoNet, an automatic hierarchical classification scheme of all protein sequences from SwissProt. Our results show that proteins that are remote from solved structures in the ProtoNet hierarchy are more likely to belong to new superfamilies. The results are validated against SCOP releases from recent years that account for about half of the solved structures known to date. We show that our new method and the representation of ProtoNet are superior in detecting new targets, compared to our previous method using ProtoMap classification. Furthermore, our method outperforms PSI-BLAST search in detecting potential new superfamilies. PMID- 15539448 TI - Primaclade--a flexible tool to find conserved PCR primers across multiple species. AB - Primaclade is a web-based application that accepts a multiple species nucleotide alignment file as input and identifies a set of polymerase chain reaction (PCR) primers that will bind across the alignment. Primaclade iteratively runs the Primer3 application for each alignment sequence and collates the results. Primaclade creates an HTML results page that recaps the original alignment, provides a consensus sequence and lists primers for each alignment area, with primers color-coded to reflect the level of degeneracy in the primer. PMID- 15539449 TI - Detection of low level genomic alterations by comparative genomic hybridization based on cDNA micro-arrays. AB - MOTIVATION: The accumulation of genomic alterations is an important process in tumor formation and progression. Comparative genomic hybridization performed on cDNA arrays (cDNA aCGH) is a common method to investigate the genomic alterations on a genome-wide scale. However, when detecting low-level DNA copy number changes this technology requires the use of noise reduction strategies due to a low signal to noise ratio. RESULTS: Currently a running average smoothing filter is the most frequently used noise reduction strategy. We analyzed this strategy theoretically and experimentally and found that it is not sensitive to very low level genomic alterations. The presence of systematic errors in the data is one of the main reasons for this failure. We developed a novel algorithm which efficiently reduces systematic noise and allows for the detection of low-level genomic alterations. The algorithm is based on comparison of the biological relevant data to data from so-called self-self hybridizations, additional experiments which contain no biological information but contain systematic errors. We find that with our algorithm the effective resolution for +/-1 DNA copy number changes is about 2 Mb. For copy number changes larger than three the effective resolution is on the level of single genes. PMID- 15539450 TI - NetAcet: prediction of N-terminal acetylation sites. AB - We present here a neural network based method for prediction of N-terminal acetylation-by far the most abundant post-translational modification in eukaryotes. The method was developed on a yeast dataset for N-acetyltransferase A (NatA) acetylation, which is the type of N-acetylation for which most examples are known and for which orthologs have been found in several eukaryotes. We obtain correlation coefficients close to 0.7 on yeast data and a sensitivity up to 74% on mammalian data, suggesting that the method is valid for eukaryotic NatA orthologs. PMID- 15539451 TI - A parallel graph decomposition algorithm for DNA sequencing with nanopores. AB - MOTIVATION: With the potential availability of nanopore devices that can sense the bases of translocating single-stranded DNA (ssDNA), it is likely that 'reads' of length approximately 10(5) will be available in large numbers and at high speed. We address the problem of complete DNA sequencing using such reads. We assume that approximately 10(2) copies of a DNA sequence are split into single strands that break into randomly sized pieces as they translocate the nanopore in arbitrary orientations. The nanopore senses and reports each individual base that passes through, but all information about orientation and complementarity of the ssDNA subsequences is lost. Random errors (both biological and transduction) in the reads create further complications. RESULTS: We have developed an algorithm that addresses these issues. It can be considered an extreme variation of the well-known Eulerian path approach. It searches over a space of de Bruijn graphs until it finds one in which (a) the impact of errors is eliminated and (b) both possible orientations of the two ssDNA sequences can be identified separately and unambiguously. Our algorithm is able to correctly reconstruct real DNA sequences of the order of 10(6) bases (e.g. the bacterium Mycoplasma pneumoniae) from simulated erroneous reads on a modest workstation in about 1 h. We describe, and give measured timings of, a parallel implementation of this algorithm on the Cray Multithreaded Architecture (MTA-2) supercomputer, whose architecture is ideally suited to this 'unstructured' problem. Our parallel implementation is crucial to the problem of rapidly sequencing long DNA sequences and also to the situation where multiple nanopores are used to obtain a high-bandwidth stream of reads. PMID- 15539452 TI - An improved algorithm for stoichiometric network analysis: theory and applications. AB - MOTIVATION: Genome scale analysis of the metabolic network of a microorganism is a major challenge in bioinformatics. The combinatorial explosion, which occurs during the construction of elementary fluxes (non-redundant pathways) requires sophisticated and efficient algorithms to tackle the problem. RESULTS: Mathematically, the calculation of elementary fluxes amounts to characterizing the space of solutions to a mixed system of linear equalities, given by the stoichiometry matrix, and linear inequalities, arising from the irreversibility of some or all of the reactions in the network. Previous approaches to this problem have iteratively solved for the equalities while satisfying the inequalities throughout the process. In an extension of previous work, here we consider the complementary approach and derive an algorithm which satisfies the inequalities one by one while staying in the space of solution of the equality constraints. Benchmarks on different subnetworks of the central carbon metabolism of Escherichia coli show that this new approach yields a significant reduction in the execution time of the calculation. This reduction arises since the odds that an intermediate elementary flux already fulfills an additional inequality are larger than when having to satisfy an additional equality constraint. PMID- 15539453 TI - Operon prediction without a training set. AB - MOTIVATION: Annotation of operons in a bacterial genome is an important step in determining an organism's transcriptional regulatory program. While extensive studies of operon structure have been carried out in a few species such as Escherichia coli, fewer resources exist to inform operon prediction in newly sequenced genomes. In particular, many extant operon finders require a large body of training examples to learn the properties of operons in the target organism. For newly sequenced genomes, such examples are generally not available; moreover, a model of operons trained on one species may not reflect the properties of other, distantly related organisms. We encountered these issues in the course of predicting operons in the genome of Bacteroides thetaiotaomicron (B.theta), a common anaerobe that is a prominent component of the normal adult human intestinal microbial community. RESULTS: We describe an operon predictor designed to work without extensive training data. We rely on a small set of a priori assumptions about the properties of the genome being annotated that permit estimation of the probability that two adjacent genes lie in a common operon. Predictions integrate several sources of information, including intergenic distance, common functional annotation and a novel formulation of conserved gene order. We validate our predictor both on the known operons of E.coli and on the genome of B.theta, using expression data to evaluate our predictions in the latter. PMID- 15539454 TI - TMDET: web server for detecting transmembrane regions of proteins by using their 3D coordinates. AB - The structure of integral membrane proteins is determined in the absence of the lipid bilayer; consequently the membrane localization of the protein is usually not specified in the corresponding PDB file. Recently, we have developed a new method called TMDET which determines the most possible localization of the membrane relative to the protein structure, and gives the annotation of the membrane embedded parts of the sequence. The entire Protein Data Bank has been scanned by the new TMDET algorithm resulting in the database of structurally determined transmembrane proteins (PDB_TM). Here we present the web interface of the TMDET algorithm to allow scientists to determine the membrane localization of structural data prior to deposition or to analyze model structures. PMID- 15539455 TI - The cellular prion protein modulates phagocytosis and inflammatory response. AB - The cellular prion protein (PrPc) is a glycoprotein anchored by glycosylphosphatidylinositol (GPI) to the cell surface and is abundantly expressed in the central nervous system. It is also expressed in a variety of cell types of the immune system. We investigated the role of PrPc in the phagocytosis of apoptotic cells and other particles. Macrophages from mice with deletion of the Prnp gene showed higher rates of phagocytosis than wild-type macrophages in in vitro assays. The elimination of GPI-anchored proteins from the cell surface of macrophages from wild-type mice rendered these cells as efficient as macrophages derived from knockout mice. In situ detection of phagocytosis of apoptotic bodies within the retina indicated augmented phagocytotic activity in knockout mice. In an in vivo assay of acute peritonitis, knockout mice showed more efficient phagocytosis of zymosan particles than wild-type mice. In addition, leukocyte recruitment was altered in knockout mice, as compared with wild type. The data show that PrPc modulates phagocytosis in vitro and in vivo. This activity is described for the first time and may be important for normal macrophage functions as well as for the pathogenesis of prion diseases. PMID- 15539456 TI - Transgenic expression of CCL2 in the central nervous system prevents experimental autoimmune encephalomyelitis. AB - CC chemokine ligand 2 (CCL2)/monocyte chemotactic protein-1, a member of the CC chemokine family, is a chemoattractant for monocytes and T cells through interaction with its receptor CCR2. In the present study, we examined a T helper cell type 1 (Th1)-dependent disease, proteolipid protein-induced experimental autoimmune encephalomyelitis, in a transgenic mouse line that constitutively expressed low levels of CCL2 in the central nervous system (CNS) under control of the astrocyte-specific glial fibrillary acidic protein promoter. CCL2 transgenic mice developed significantly milder clinical disease than littermate controls. As determined by flow cytometry, mononuclear cell infiltrates in the CNS tissues of CCL2 transgenic and littermate-control mice contained equal numbers of CD4+ and CD8+ T cells, and the CCL2 transgenic mice showed an enhanced number of CNS infiltrating monocytes. CNS antigen-specific T cells from CCL2 transgenic mice produced markedly less interferon-gamma. Overexpression of CCL2 in the CNS resulted in decreased interleukin-12 receptor expression by antigen-specific T cells. Collectively, these results indicate that sustained, tissue-specific expression of CCL2 in vivo down-regulates the Th1 autoimmune response, culminating in milder clinical disease. PMID- 15539457 TI - Blockade of alpha6 integrin inhibits IL-1beta- but not TNF-alpha-induced neutrophil transmigration in vivo. AB - In vitro and in vivo evidence supports a functional role for the integrin alpha6beta1 in neutrophil migration through the perivascular basement membrane, a response that in vivo appears to be associated with platelet/endothelial cell adhesion molecule-1 (PECAM-1)-mediated up-regulation of alpha6beta1 on the cell surface of transmigrating leukocytes. As the involvement of PECAM-1 in leukocyte migration is cytokine-specific, the aim of the present study was to investigate whether alpha6beta1 exhibited a similar profile of stimulus specificity in this context. The cytokines interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) were used to elicit neutrophil migration in two murine models of inflammation, migration through cremasteric venules, as observed by intravital microscopy, and migration into the peritoneal cavity. The role of alpha6beta1 was investigated using an alpha6 integrin-blocking monoclonal antibody GoH3. In both models, GoH3 significantly inhibited neutrophil transmigration induced by IL 1beta but not TNF-alpha. This cytokine-specific role of alpha6 integrin was associated with enhanced cell-surface expression of alpha6beta1 on transmigrated neutrophils (as compared with blood cells) in response to IL-1beta but not TNF alpha. Using lipopolysaccharide as an inflammatory stimulus in the cremaster muscle model, the study also provides evidence for the involvement of alpha6 integrin in leukocyte transmigration as mediated by endogenously generated IL 1beta. Collectively, the findings demonstrate that alpha6beta1 blockade inhibits neutrophil migration induced by exogenous and endogenous IL-1beta but not TNF alpha, observations that are associated with increased expression of the integrin on transmigrated leukocytes. PMID- 15539458 TI - Characterization of the interstitial lung and peripheral blood T cell receptor repertoire in cigarette smokers. AB - T lymphocytes modulate the pulmonary inflammatory response. The aim of this study was to evaluate the clonality within the interstitial lung and peripheral blood T cell receptor (TCR) repertoire in smokers. Interstitial T lymphocytes were isolated from surplus tissue of 16 patients (63 +/- 9 [+/- SD] yr old, 11 male) undergoing surgery due to lung cancer (n = 15) or emphysema. TCR clonality was assessed by PCR amplification followed by spectratyping. Nearly all TCR of interstitial lung lymphocytes showed oligoclonal bands (CD4(+) subset 13/16 patients, 81%; CD8(+) 100%) indicating a specific differentiation. Peripheral blood T lymphocytes (PBL) TCR (especially CD4(+)) had less oligoclonal bands (CD4(+) 31%, CD8(+) 88%). Likewise, more oligoclonal bands were seen in lung TCR (total of 168 bands; 37 CD4(+); 131 CD8(+)), compared with 59 bands in PBL TCR (13 CD4(+); 46 CD8(+)). Intraindividual comparison revealed a more prominent difference in TCR oligoclonality between lung and blood in CD8(+) T cells (median of difference lung minus blood 5; interquartile range 1-10; P = 0.002) compared with CD4(+) T cells (median 2, 0-3, P = 0.039). Thus, TCR oligoclonality is preferentially found in the CD8(+) T cell subset, most distinctive in the lung. These findings indicate a specific interstitial T cell differentiation in response to local stimuli. PMID- 15539459 TI - Prostaglandin E(2) inhibits fibroblast migration by E-prostanoid 2 receptor mediated increase in PTEN activity. AB - An increased migratory phenotype exists in lung fibroblasts derived from patients with fibroproliferative lung disease. Prostaglandin E(2) (PGE(2)) suppresses fibroblast migration, but the receptor(s) and mechanism(s) mediating this action are unknown. Our data confirm that treatment of human lung fibroblasts with PGE(2) inhibits migration. Similar effects of butaprost, an E-prostanoid (EP) 2 receptor-specific ligand, implicate the EP2 receptor in migration-inhibitory signaling. Further, migration in fibroblasts deficient for the EP2 receptor cannot be inhibited by PGE(2) or butaprost, confirming the central role of EP2 in mediating these effects. Our previous data suggested that phosphatase and tensin homolog on chromosome ten (PTEN), a phosphatase that opposes the actions of phosphatidylinositol-3-kinase (PI3K), may be important in regulating lung fibroblast motility. We now report that both PGE(2) and butaprost increase PTEN phosphatase activity, without a concomitant increase in PTEN protein levels. This contributes to EP2-mediated migration inhibition, because migration in PTEN-null fibroblasts is similarly unaffected by EP2 receptor signaling. Increased PTEN activity in response to EP2 stimulation is associated with decreased tyrosine phosphorylation on PTEN, a mechanism known to regulate enzyme activity. Collectively, these data describe the novel mechanistic finding that PGE(2), via the EP2 receptor, decreases tyrosine phosphorylation on PTEN, resulting in increased PTEN enzyme activity and inhibition of fibroblast migration. PMID- 15539460 TI - Mechanotransduction and strain amplification in osteocyte cell processes. AB - A paradox in bone tissue is that tissue-level strains due to animal and human locomotion are too small to initiate intracellular chemical responses directly. A model recently was proposed to resolve this paradox, which predicts that the fluid flow through the pericellular matrix in the lacunar-canalicular porosity due to mechanical loading can induce strains in the actin filament bundles of the cytoskeleton that are more than an order of magnitude larger than tissue level strains. In this study, we greatly refine this model by using the latest ultrastructural data for the cell process cytoskeleton, the tethering elements that attach the process to the canalicular wall and their finite flexural rigidity EI. We construct a much more realistic 3D model for the osteocyte process and then use large-deformation "elastica" theory for finite EI to predict the deformed shape of the tethering elements and the hoop strain on the central actin bundle. Our model predicts a cell process that is 3 times stiffer than in a previous study but hoop strain of >0.5% for tissue-level strains of >1,000 microstrain at 1 Hz and >250 microstrain at frequencies >10 Hz. We propose that this strain-amplification model provides a more likely hypothesis for the excitation of osteocytes than the previously proposed fluid-shear hypothesis. PMID- 15539461 TI - Finding new components of the target of rapamycin (TOR) signaling network through chemical genetics and proteome chips. AB - The TOR (target of rapamycin) proteins play important roles in nutrient signaling in eukaryotic cells. Rapamycin treatment induces a state reminiscent of the nutrient starvation response, often resulting in growth inhibition. Using a chemical genetic modifier screen, we identified two classes of small molecules, small-molecule inhibitors of rapamycin (SMIRs) and small-molecule enhancers of rapamycin (SMERs), that suppress and augment, respectively, rapamycin's effect in the yeast Saccharomyces cerevisiae. Probing proteome chips with biotinylated SMIRs revealed putative intracellular target proteins, including Tep1p, a homolog of the mammalian PTEN (phosphatase and tensin homologue deleted on chromosome 10) tumor suppressor, and Ybr077cp (Nir1p), a protein of previously unknown function that we show to be a component of the TOR signaling network. Both SMIR target proteins are associated with PI(3,4)P2, suggesting a mechanism of regulation of the TOR pathway involving phosphatidylinositides. Our results illustrate the combined use of chemical genetics and proteomics in biological discovery and map a path for creating useful therapeutics for treating human diseases involving the TOR pathway, such as diabetes and cancer. PMID- 15539462 TI - Mimicking enzyme evolution by generating new (betaalpha)8-barrels from (betaalpha)4-half-barrels. AB - Gene duplication and fusion events that multiply and link functional protein domains are crucial mechanisms of enzyme evolution. The analysis of amino acid sequences and three-dimensional structures suggested that the (betaalpha)8 barrel, which is the most frequent fold among enzymes, has evolved by the duplication, fusion, and mixing of (betaalpha)4-half-barrel domains. Here, we mimicked this evolutionary strategy by generating in vitro (betaalpha)8-barrels from (betaalpha)4-half-barrels that were deduced from the enzymes imidazole glycerol phosphate synthase (HisF) and N'[(5'-phosphoribosyl)formimino]-5 aminoimidazole-4-carboxamide-ribonucleotide isomerase (HisA). To this end, the gene for the C-terminal (betaalpha)4-half-barrel (HisF-C) of HisF was duplicated and fused in tandem to yield HisF-CC, which is more stable than HisF-C. In the next step, by optimizing side-chain interactions within the center of the beta barrel of HisF-CC, the monomeric and compact (betaalpha)8-barrel protein HisF-C*C was generated. Moreover, the genes for the N- and C-terminal (betaalpha)4-half barrels of HisF and HisA were fused crosswise to yield the chimeric proteins HisFA and HisAF. Whereas HisFA contains native secondary structure elements but adopts ill-defined association states, the (betaalpha)8-barrel HisAF is a stable and compact monomer that reversibly unfolds with high cooperativity. The results obtained suggest a previously undescribed dimension for the diversification of enzymatic activities: new (betaalpha)8-barrels with novel functions might have evolved by the exchange of (betaalpha)4-half-barrel domains with distinct functional properties. PMID- 15539463 TI - Bbs2-null mice have neurosensory deficits, a defect in social dominance, and retinopathy associated with mislocalization of rhodopsin. AB - Bardet-Biedl syndrome (BBS) is a heterogeneous, pleiotropic human disorder characterized by obesity, retinopathy, polydactyly, renal and cardiac malformations, learning disabilities, hypogenitalism, and an increased incidence of diabetes and hypertension. No information is available regarding the specific function of BBS2. We show that mice lacking Bbs2 gene expression have major components of the human phenotype, including obesity and retinopathy. In addition, these mice have phenotypes associated with cilia dysfunction, including retinopathy, renal cysts, male infertility, and a deficit in olfaction. With the exception of male infertility, these phenotypes are not caused by a complete absence of cilia. We demonstrate that BBS2 retinopathy involves normal retina development followed by apoptotic death of photoreceptors, the primary ciliated cells of the retina. Photoreceptor cell death is preceded by mislocalization of rhodopsin, indicating a defect in transport. We also demonstrate that Bbs2(-/-) mice and a second BBS mouse model, Bbs4(-/-), have a defect in social function. The evaluation of Bbs2(-/-) mice indicates additional phenotypes that should be evaluated in human patients, including deficits in social interaction and infertility. PMID- 15539464 TI - A radical approach is needed to eliminate interference from endogenous antibodies in immunoassays. PMID- 15539465 TI - Effects of processing and storage conditions on amyloid beta (1-42) and tau concentrations in cerebrospinal fluid: implications for use in clinical practice. AB - BACKGROUND: Reported concentrations of amyloid beta (1-42) (A beta 42) and tau in cerebrospinal fluid (CSF) differ among reports. We investigated the effects of storage temperature, repeated freeze/thaw cycles, and centrifugation on the concentrations of A beta 42 and tau in CSF. METHODS: Stability of samples stored at -80 degrees C was determined by use of an accelerated stability testing protocol according to the Arrhenius equation. A beta 42 and tau concentrations were measured in CSF samples stored at 4, 18, 37, and -80 degrees C. Relative CSF concentrations (%) of the biomarkers after one freeze/thaw cycle were compared with those after two, three, four, five, and six freeze/thaw cycles. In addition, relative A beta 42 and tau concentrations in samples not centrifuged were compared with samples centrifuged after 1, 4, 48, and 72 h. RESULTS: A beta 42 and tau concentrations were stable in CSF when stored for a long period at -80 degrees C. CSF A beta 42 decreased by 20% during the first 2 days at 4, 18, and 37 degrees C compared with -80 degrees C. CSF tau decreased after storage for 12 days at 37 degrees C. After three freeze/thaw cycles, CSF A beta 42 decreased 20%. CSF tau was stable during six freeze/thaw cycles. Centrifugation did not influence the biomarker concentrations. CONCLUSIONS: Repeated freeze/thaw cycles and storage at 4, 18, and 37 degrees C influence the quantitative result of the A beta 42 test. Preferably, samples should be stored at -80 degrees C immediately after collection. PMID- 15539466 TI - Quantitative analysis of circulating plasma DNA as a tumor marker in thoracic malignancies. AB - BACKGROUND: Increased plasma DNA has been found in cancer patients and may have potential as a tumor marker. The objectives of this study were to develop a controlled, quantitative PCR (QPCR) assay to measure plasma DNA and then evaluate plasma DNA concentrations as a tumor marker in patients with thoracic malignancies. METHODS: We developed a QPCR assay for DNA, using the human beta actin gene. Plasma samples were analyzed from 58 patients with esophageal cancer (EC; 20 banked samples and 38 prospectively collected samples) and 25 patients with lung cancer (LC; all prospectively collected). Control groups consisting of 51 patients with gastroesophageal reflux disease (GERD; 23 banked samples and 28 prospectively collected) and 11 healthy volunteers were also analyzed. RESULTS: The assay had an experimental variability <4%. In our banked samples, the mean concentration of plasma DNA in EC was 819.0 microg/L (range, 46.2-4738.0 microg/L) vs 432.0 microg/L (6.0-2888.0 microg/L) in GERD (P = 0.02). However, the prospectively collected samples had lower DNA concentrations, and there was no difference between cancer patients and controls. The mean DNA concentration was 10.6 microg/L (range, 7.0-14.0 microg/L) in healthy volunteers and 10.5 microg/L (range, 4.0-23.5 microg/L) in GERD controls vs 13.0 microg/L (range, 4.5 46.5 microg/L) in EC and 14.6 microg/L (range, 3.0-30.0 microg/L) in LC. CONCLUSIONS: Our data indicate that plasma DNA concentrations are of limited diagnostic value when samples are prospectively collected and uniformly handled. This is in contrast to previously published results. Qualitative analysis of DNA may be needed if plasma nucleic acids are to be used as a diagnostic tool in cancer screening. PMID- 15539467 TI - Serum lipid concentrations and prevalence of dyslipidemia in a large professional population in Beijing. AB - BACKGROUND: Lipid abnormalities are major risk factors for premature coronary artery diseases. We investigated serum lipids and the prevalence of dyslipidemia in a professional population in Beijing and compared these data with those obtained in a similar population during 1984-1986. METHODS: We studied 14 963 individuals 20-90 years of age. Health status was determined by questionnaires and physical check-ups. Total cholesterol (TC), LDL-cholesterol (LDL-C), HDL cholesterol (HDL-C), and total triglycerides (TGs) were measured. TC >5.7 mmol/L, LDL-C >3.6 mmol/L, TGs >1.7 mmol/L, and HDL-C <0.9 mmol/L were defined as abnormal. RESULTS: Mean serum TC, LDL-C, and TG concentrations were increased compared with the values obtained in 1984-1986, with 52.7% of males and 42.9% of females having at least one abnormal lipid concentration. Hypercholesterolemia occurred in 6% of males and 2.8% of females in the younger group (20-39 years) and in 20.2% of males and 38.7% of females in the older group (>60 years). HDL-C was abnormally low in approximately 7% of males and in 1.6% of females. The prevalences of hypercholesterolemia, hypertriglyceridemia, and abnormally low HDL C, especially the presence of slight hypertriglyceridemia, were higher than in 1984-1986 in all age groups. The increase was most prominent in the middle age group (40-59 years). CONCLUSIONS: Hypercholesterolemia, hypertriglyceridemia, and abnormally low HDL-C have increased considerably over the past 20 years in professional populations in Beijing. Dietary changes and less physical activity resulting from rapid improvements in living conditions may be the causes for the increases. Enhanced preventive measures should be undertaken to modify these situations. PMID- 15539468 TI - FRAGILE FIBER3, an Arabidopsis gene encoding a type II inositol polyphosphate 5 phosphatase, is required for secondary wall synthesis and actin organization in fiber cells. AB - Type II inositol polyphosphate 5-phosphatases (5PTases) in yeast and animals have been known to regulate the level of phosphoinositides and thereby influence various cellular activities, such as vesicle trafficking and actin organization. In plants, little is known about the phosphatases involved in hydrolysis of phosphoinositides, and roles of type II 5PTases in plant cellular functions have not yet been characterized. In this study, we demonstrate that the FRAGILE FIBER3 (FRA3) gene of Arabidopsis thaliana, which encodes a type II 5PTase, plays an essential role in the secondary wall synthesis in fiber cells and xylem vessels. The fra3 mutations caused a dramatic reduction in secondary wall thickness and a concomitant decrease in stem strength. These phenotypes were associated with an alteration in actin organization in fiber cells. Consistent with the defective fiber and vessel phenotypes, the FRA3 gene was found to be highly expressed in fiber cells and vascular tissues in stems. The FRA3 protein is composed of two domains, an N-terminal localized WD-repeat domain and a C-terminal localized 5PTase catalytic domain. In vitro activity assay demonstrated that recombinant FRA3 exhibited phosphatase activity toward PtdIns(4,5)P2, PtdIns(3,4,5)P3, and Ins(1,4,5)P3, with the highest substrate affinity toward PtdIns(4,5)P2. The fra3 missense mutation, which caused an amino acid substitution in the conserved motif II of the 5PTase catalytic domain, completely abolished the FRA3 phosphatase activity. Moreover, the endogenous levels of PtdIns(4,5)2 and Ins(1,4,5)P3 were found to be elevated in fra3 stems. Together, our findings suggest that the FRA3 type II 5PTase is involved in phosphoinositide metabolism and influences secondary wall synthesis and actin organization. PMID- 15539469 TI - The vegetative vacuole proteome of Arabidopsis thaliana reveals predicted and unexpected proteins. AB - Vacuoles play central roles in plant growth, development, and stress responses. To better understand vacuole function and biogenesis we have characterized the vegetative vacuolar proteome from Arabidopsis thaliana. Vacuoles were isolated from protoplasts derived from rosette leaf tissue. Total purified vacuolar proteins were then subjected either to multidimensional liquid chromatography/tandem mass spectrometry or to one-dimensional SDS-PAGE coupled with nano-liquid chromatography/tandem mass spectrometry (nano-LC MS/MS). To ensure maximum coverage of the proteome, a tonoplast-enriched fraction was also analyzed separately by one-dimensional SDS-PAGE followed by nano-LC MS/MS. Cumulatively, 402 proteins were identified. The sensitivity of our analyses is indicated by the high coverage of membrane proteins. Eleven of the twelve known vacuolar-ATPase subunits were identified. Here, we present evidence of four tonoplast-localized soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), representing each of the four groups of SNARE proteins necessary for membrane fusion. In addition, potential cargo of the N- and C terminal propeptide sorting pathways, association of the vacuole with the cytoskeleton, and the vacuolar localization of 89 proteins of unknown function are identified. A detailed analysis of these proteins and their roles in vacuole function and biogenesis is presented. PMID- 15539470 TI - Encounters between dynamic cortical microtubules promote ordering of the cortical array through angle-dependent modifications of microtubule behavior. AB - Ordered cortical microtubule arrays are essential for normal plant morphogenesis, but how these arrays form is unclear. The dynamics of individual cortical microtubules are stochastic and cannot fully account for the observed order; however, using tobacco (Nicotiana tabacum) cells expressing either the MBD-DsRed (microtubule binding domain of the mammalian MAP4 fused to the Discosoma sp red fluorescent protein) or YFP-TUA6 (yellow fluorescent protein fused to the Arabidopsis alpha-tubulin 6 isoform) microtubule markers, we identified intermicrotubule interactions that modify their stochastic behaviors. The intermicrotubule interactions occur when the growing plus-ends of cortical microtubules encounter previously existing cortical microtubules. Importantly, the outcome of such encounters depends on the angle at which they occur: steep angle collisions are characterized by approximately sevenfold shorter microtubule contact times compared with shallow-angle encounters, and steep-angle collisions are twice as likely to result in microtubule depolymerization. Hence, steep-angle collisions promote microtubule destabilization, whereas shallow-angle encounters promote both microtubule stabilization and coalignment. Monte Carlo modeling of the behavior of simulated microtubules, according to the observed behavior of transverse and longitudinally oriented cortical microtubules in cells, reveals that these simple rules for intermicrotubule interactions are necessary and sufficient to facilitate the self-organization of dynamic microtubules into a parallel configuration. PMID- 15539471 TI - Characterization of antirrhinum petal development and identification of target genes of the class B MADS box gene DEFICIENS. AB - The class B MADS box transcription factors DEFICIENS (DEF) and GLOBOSA (GLO) of Antirrhinum majus together control the organogenesis of petals and stamens. Toward an understanding of how the downstream molecular mechanisms controlled by DEF contribute to petal organogenesis, we conducted expression profiling experiments using macroarrays comprising >11,600 annotated Antirrhinum unigenes. First, four late petal developmental stages were compared with sepals. More than 500 ESTs were identified that comprise a large number of stage-specifically regulated genes and reveal a highly dynamic transcriptional regulation. For identification of DEF target genes that might be directly controlled by DEF, we took advantage of the temperature-sensitive def-101 mutant. To enhance the sensitivity of the profiling experiments, one petal developmental stage was selected, characterized by increased transcriptome changes that reflect the onset of cell elongation processes replacing cell division processes. Upon reduction of the DEF function, 49 upregulated and 52 downregulated petal target genes were recovered. Eight target genes were further characterized in detail by RT-PCR and in situ studies. Expression of genes responding rapidly toward an altered DEF activity is confined to different petal tissues, demonstrating the complexity of the DEF function regulating diverse basic processes throughout petal morphogenesis. PMID- 15539472 TI - Phosphorylation of 1-aminocyclopropane-1-carboxylic acid synthase by MPK6, a stress-responsive mitogen-activated protein kinase, induces ethylene biosynthesis in Arabidopsis. AB - Mitogen-activated protein kinases (MAPKs) are implicated in regulating plant growth, development, and response to the environment. However, the underlying mechanisms are unknown because of the lack of information about their substrates. Using a conditional gain-of-function transgenic system, we demonstrated that the activation of SIPK, a tobacco (Nicotiana tabacum) stress-responsive MAPK, induces the biosynthesis of ethylene. Here, we report that MPK6, the Arabidopsis thaliana ortholog of tobacco SIPK, is required for ethylene induction in this transgenic system. Furthermore, we found that selected isoforms of 1-aminocyclopropane-1 carboxylic acid synthase (ACS), the rate-limiting enzyme of ethylene biosynthesis, are substrates of MPK6. Phosphorylation of ACS2 and ACS6 by MPK6 leads to the accumulation of ACS protein and, thus, elevated levels of cellular ACS activity and ethylene production. Expression of ACS6(DDD), a gain-of-function ACS6 mutant that mimics the phosphorylated form of ACS6, confers constitutive ethylene production and ethylene-induced phenotypes. Increasing numbers of stress stimuli have been shown to activate Arabidopsis MPK6 or its orthologs in other plant species. The identification of the first plant MAPK substrate in this report reveals one mechanism by which MPK6/SIPK regulates plant stress responses. Equally important, this study uncovers a signaling pathway that modulates the biosynthesis of ethylene, an important plant hormone, in plants under stress. PMID- 15539473 TI - The essential basic helix-loop-helix protein FIT1 is required for the iron deficiency response. AB - Regulation of iron uptake is critical for plant survival. Although the activities responsible for reduction and transport of iron at the plant root surface have been described, the genes controlling these activities are largely unknown. We report the identification of the essential gene Fe-deficiency Induced Transcription Factor 1 (FIT1), which encodes a putative transcription factor that regulates iron uptake responses in Arabidopsis thaliana. Like the Fe(III) chelate reductase FRO2 and high affinity Fe(II) transporter IRT1, FIT1 mRNA is detected in the outer cell layers of the root and accumulates in response to iron deficiency. fit1 mutant plants are chlorotic and die as seedlings but can be rescued by the addition of supplemental iron, pointing to a defect in iron uptake. fit1 mutant plants accumulate less iron than wild-type plants in root and shoot tissues. Microarray analysis shows that expression of many (72 of 179) iron regulated genes is dependent on FIT1. We demonstrate that FIT1 regulates FRO2 at the level of mRNA accumulation and IRT1 at the level of protein accumulation. We propose a new model for iron uptake in Arabidopsis where FRO2 and IRT1 are differentially regulated by FIT1. PMID- 15539483 TI - Coupling segmentation to axis formation. AB - A characteristic feature of the vertebrate body is its segmentation along the anteroposterior axis, as illustrated by the repetition of vertebrae that form the vertebral column. The vertebrae and their associated muscles derive from metameric structures of mesodermal origin, the somites. The segmentation of the body is established by somitogenesis, during which somites form sequentially in a rhythmic fashion from the presomitic mesoderm. This review highlights recent findings that show how dynamic gradients of morphogens and retinoic acid, coupled to a molecular oscillator, drive the formation of somites and link somitogenesis to the elongation of the anteroposterior axis. PMID- 15539484 TI - Epsin potentiates Notch pathway activity in Drosophila and C. elegans. AB - Endocytosis and trafficking within the endocytosis pathway are known to modulate the activity of different signaling pathways. Epsins promote endocytosis and are postulated to target specific proteins for regulated endocytosis. Here, we present a functional link between the Notch pathway and epsins. We identify the Drosophila ortholog of epsin, liquid facets (lqf), as an inhibitor of cardioblast development in a genetic screen for mutants that affect heart development. We find that lqf inhibits cardioblast development and promotes the development of fusion-competent myoblasts, suggesting a model in which lqf acts on or in fusion competent myoblasts to prevent their acquisition of the cardioblast fate. lqf and Notch exhibit essentially identical heart phenotypes, and lqf genetically interacts with the Notch pathway during multiple Notch-dependent events in Drosophila. We extended the link between the Notch pathway and epsin function to C. elegans, where the C. elegans lqf ortholog acts in the signaling cell to promote the glp-1/Notch pathway activity during germline development. Our results suggest that epsins play a specific, evolutionarily conserved role to promote Notch signaling during animal development and support the idea that they do so by targeting ligands of the Notch pathway for endocytosis. PMID- 15539485 TI - Altered endochondral bone development in matrix metalloproteinase 13-deficient mice. AB - The assembly and degradation of extracellular matrix (ECM) molecules are crucial processes during bone development. In this study, we show that ECM remodeling is a critical rate-limiting step in endochondral bone formation. Matrix metalloproteinase (MMP) 13 (collagenase 3) is poised to play a crucial role in bone formation and remodeling because of its expression both in terminal hypertrophic chondrocytes in the growth plate and in osteoblasts. Moreover, a mutation in the human MMP13 gene causes the Missouri variant of spondyloepimetaphyseal dysplasia. Inactivation of Mmp13 in mice through homologous recombination led to abnormal skeletal growth plate development. Chondrocytes differentiated normally but their exit from the growth plate was delayed. The severity of the Mmp13- null growth plate phenotype increased until about 5 weeks and completely resolved by 12 weeks of age. Mmp13-null mice had increased trabecular bone, which persisted for months. Conditional inactivation of Mmp13 in chondrocytes and osteoblasts showed that increases in trabecular bone occur independently of the improper cartilage ECM degradation caused by Mmp13 deficiency in late hypertrophic chondrocytes. Our studies identified the two major components of the cartilage ECM, collagen type II and aggrecan, as in vivo substrates for MMP13. We found that degradation of cartilage collagen and aggrecan is a coordinated process in which MMP13 works synergistically with MMP9. Mice lacking both MMP13 and MMP9 had severely impaired endochondral bone, characterized by diminished ECM remodeling, prolonged chondrocyte survival, delayed vascular recruitment and defective trabecular bone formation (resulting in drastically shortened bones). These data support the hypothesis that proper ECM remodeling is the dominant rate-limiting process for programmed cell death, angiogenesis and osteoblast recruitment during normal skeletal morphogenesis. PMID- 15539486 TI - Par-1 regulates bicoid mRNA localisation by phosphorylating Exuperantia. AB - The Ser/Thr kinase Par-1 is required for cell polarisation in diverse organisms such as yeast, worms, flies and mammals. During Drosophila oogenesis, Par-1 is required for several polarisation events, including localisation of the anterior determinant bicoid. To elucidate the molecular pathways triggered by Par-1, we have performed a genome-wide, high-throughput screen for Par-1 targets. Among the targets identified in this screen was Exuperantia (Exu), a mediator of bicoid mRNA localisation. We show that Exu is a phosphoprotein whose phosphorylation is dependent on Par-1 in vitro and in vivo. We identify two motifs in Exu that are phosphorylated by Par-1, and show that their mutation abolishes bicoid mRNA localisation during mid-oogenesis. Interestingly, exu mutants in which Exu phosphorylation is specifically affected can to some extent recover from these bicoid mRNA localisation defects during late oogenesis. These results demonstrate that Par-1 establishes polarity in the oocyte by activating a mediator of bicoid mRNA localisation. Furthermore, our analysis reveals two phases of Exu-dependent bicoid mRNA localisation: an early phase that is strictly dependent on Exu phosphorylation and a late phase that is less phosphorylation dependent. PMID- 15539487 TI - Zebrafish Dapper1 and Dapper2 play distinct roles in Wnt-mediated developmental processes. AB - Wnt signaling pathways in vertebrates use the phosphoprotein Dishevelled (Dvl). The cellular responses to Wnt signaling may in part be modulated by Dvl associated proteins, including Dapper (Dpr). We have cloned and characterized the zebrafish Dpr paralogs Dpr1 and Dpr2. Loss-of-function studies reveal that endogenous Dpr1 but not Dpr2 is required to enhance Wnt/beta-catenin activity in zebrafish embryos that are hypomorphic for Wnt8. Conversely, Dpr2 but not Dpr1 is required for normal convergence extension movements in embryos that are hypomorphic for Stbm or Wnt11, supporting a functional interaction of Dpr2 with Wnt/Ca2+-PCP signaling. In gain-of-function experiments, Dpr1 but not Dpr2 induces Wnt/beta-catenin target genes. Dpr1 synergizes with zebrafish Dvl2, and with the Dvl-interacting kinases CK1epsilon, Par1 and CK2, in activating target genes. We conclude that two Dvl-associated paralogs, Dpr1 and Dpr2, participate in distinct Wnt-dependent developmental processes. PMID- 15539489 TI - mau-2 acts cell-autonomously to guide axonal migrations in Caenorhabditis elegans. AB - The gene mau-2 has been found to be required for the guidance of cellular and axonal migrations along both the anteroposterior and the dorsoventral body axes during the development of the nematode C. elegans. We show that mau-2 encodes a novel, previously uncharacterized protein that is highly conserved among animals. Maternal mau-2 gene expression is sufficient for normal development until the fourth larval stage, and a MAU-2::GFP fusion protein localizes to the cytoplasm of neurones. mau-2 is ubiquitously expressed in embryos by late gastrulation and becomes predominantly expressed in the nervous system as morphogenesis progresses. Expression of mau-2 within individual neurones rescues the guidance defects of mau-2 mutants, indicating that mau-2 functions cell-autonomously. Altering the activity of both the dorsal repellent slt-1 and mau-2 leads to the abnormal dorsal projection of the AVM axon, a phenotype that is novel and specific to the interaction of these two genes, indicating that mau-2 participates in the guidance of AVM by a slt-1-independent mechanism. Taken together, mau-2 defines a novel guidance factor that might be involved in the intracellular processing of guidance cues encountered by migrating cells and axons during development. PMID- 15539488 TI - Inhibition of Wnt/Axin/beta-catenin pathway activity promotes ventral CNS midline tissue to adopt hypothalamic rather than floorplate identity. AB - Ventral midline cells in the neural tube form floorplate throughout most of the central nervous system (CNS) but in the anterior forebrain, they differentiate with hypothalamic identity. The signalling pathways responsible for subdivision of midline neural tissue into hypothalamic and floorplate domains are uncertain, and in this study, we have explored the role of the Wnt/Axin/beta-catenin pathway in this process. This pathway has been implicated in anteroposterior regionalisation of the dorsal neural tube but its role in patterning ventral midline tissue has not been rigorously assessed. We find that masterblind zebrafish embryos that carry a mutation in Axin1, an intracellular negative regulator of Wnt pathway activity, show an expansion of prospective floorplate coupled with a reduction of prospective hypothalamic tissue. Complementing this observation, transplantation of cells overexpressing axin1 into the prospective floorplate leads to induction of hypothalamic gene expression and suppression of floorplate marker gene expression. Axin1 is more efficient at inducing hypothalamic markers than several other Wnt pathway antagonists, and we present data suggesting that this may be due to an ability to promote Nodal signalling in addition to suppressing Wnt activity. Indeed, extracellular Wnt antagonists can promote hypothalamic gene expression when co-expressed with a modified form of Madh2 that activates Nodal signalling. These results suggest that Nodal signalling promotes the ability of cells to incorporate into ventral midline tissue, and within this tissue, antagonism of Wnt signalling promotes the acquisition of hypothalamic identity. Wnt signalling also affects patterning within the hypothalamus, suggesting that this pathway is involved in both the initial anteroposterior subdivision of ventral CNS midline fates and in the subsequent regionalisation of the hypothalamus. We suggest that by regulating the response of midline cells to signals that induce ventral fates, Axin1 and other modulators of Wnt pathway activity provide a mechanism by which cells can integrate dorsoventral and anteroposterior patterning information. PMID- 15539490 TI - Spatial and temporal regulation of ventral spinal cord precursor specification by Hedgehog signaling. AB - Graded Hedgehog (Hh) signaling patterns the spinal cord dorsoventral axis by inducing and positioning distinct precursor domains, each of which gives rise to a different type of neuron. These domains also generate glial cells, but the full range of cell types that any one precursor population produces and the mechanisms that diversify cell fate are unknown. By fate mapping and clonal analysis in zebrafish, we show that individual ventral precursor cells that express olig2 can form motoneurons, interneurons and oligodendrocytes. However, olig2+ precursors are not developmentally equivalent, but instead produce subsets of progeny cells in a spatially and temporally biased manner. Using genetic and pharmacological manipulations, we provide evidence that these biases emerge from Hh acting over time to set, maintain, subdivide and enlarge the olig2+ precursor domain and subsequently specify oligodendrocyte development. Our studies show that spatial and temporal differences in Hh signaling within a common population of neural precursors can contribute to cell fate diversification. PMID- 15539491 TI - A role for extra macrochaetae downstream of Notch in follicle cell differentiation. AB - The Drosophila ovary provides a model system for studying the mechanisms that regulate the differentiation of somatic stem cells into specific cell types. Ovarian somatic stem cells produce follicle cells, which undergo a binary choice during early differentiation. They can become either epithelial cells that surround the germline to form an egg chamber ('main body cells') or a specialized cell lineage found at the poles of egg chambers. This lineage goes on to make two cell types: polar cells and stalk cells. To better understand how this choice is made, we carried out a screen for genes that affect follicle cell fate specification or differentiation. We identified extra macrochaetae (emc), which encodes a helix-loop-helix protein, as a downstream effector of Notch signaling in the ovary. EMC is expressed in proliferating cells in the germarium, as well as in the main body follicle cells. EMC expression in the main body cells is Notch dependent, and emc mutant cells located on the main body failed to differentiate. EMC expression is reduced in the precursors of the polar and stalk cells, and overexpression of EMC caused dramatic egg chamber fusions, indicating that EMC is a negative regulator of polar and/or stalk cells. EMC and Notch were both required in the main body cells for expression of Eyes Absent (EYA), a negative regulator of polar and stalk cell fate. We propose that EMC functions downstream of Notch and upstream of EYA to regulate main body cell fate specification and differentiation. PMID- 15539492 TI - INCOMPOSITA: a MADS-box gene controlling prophyll development and floral meristem identity in Antirrhinum. AB - INCOMPOSITA (INCO) is a MADS-box transcription factor and member of the functionally diverse StMADS11 clade of the MADS-box family. The most conspicuous feature of inco mutant flowers are prophylls initiated prior to first whorl sepals at lateral positions of the flower primordium. The developing prophylls physically interfere with subsequent floral organ development that results in aberrant floral architecture. INCO, which is controlled by SQUAMOSA, prevents prophyll formation in the wild type, a role that is novel among MADS-box proteins, and we discuss evolutionary implications of this function. Overexpression of INCO or SVP, a structurally related Arabidopsis MADS-box gene involved in the negative control of Arabidopsis flowering time, conditions delayed flowering in transgenic plants, suggesting that SVP and INCO have functions in common. Enhanced flowering of squamosa mutants in the inco mutant background corroborates this potential role of INCO as a floral repressor in Antirrhinum. One further, hitherto hidden, role of INCO is the positive control of Antirrhinum floral meristem identity. This is revealed by genetic interactions between inco and mutants of FLORICAULA, a gene that controls the inflorescence to floral transition, together with SQUAMOSA. The complex regulatory and combinatorial relations between INCO, FLORICAULA and SQUAMOSA are summarised in a model that integrates observations from molecular studies as well as analyses of expression patterns and genetic interactions. PMID- 15539493 TI - The conserved kinase UNC-51 acts with VAB-8 and UNC-14 to regulate axon outgrowth in C. elegans. AB - Directional cues guide growth cones. While molecules like UNC-6/netrin direct migrations along the dorsoventral axis of many organisms, it is unclear how anteroposterior guidance is achieved. We describe a physical interaction between VAB-8, a protein both necessary and sufficient for posteriorly directed migrations in C. elegans, and UNC-51, a conserved serine/threonine kinase that functions generally in axon outgrowth. We show that both proteins function in the CAN neurons to direct their axons posteriorly. Expression in the CANs of peptides predicted to interfere with interactions between UNC-51 and both VAB-8 and UNC 14, a second protein that interacts physically with UNC-51, disrupts CAN axon outgrowth. We provide genetic evidence that VAB-8 functions in an UNC-51 pathway for posteriorly directed CAN axon guidance and show that VAB-8 and UNC-14 can be targets of UNC-51 kinase activity. Taken together, our results suggest that VAB-8 and UNC-14 are substrates that mediate the function of UNC-51 in axon outgrowth. PMID- 15539494 TI - A conserved metalloprotease mediates ecdysis in Caenorhabditis elegans. AB - Molting is required for progression between larval stages in the life cycle of nematodes. We have identified four mutant alleles of a Caenorhabditis elegans metalloprotease gene, nas-37, that cause incomplete ecdysis. At each molt the cuticle fails to open sufficiently at the anterior end and the partially shed cuticle is dragged behind the animal. The gene is expressed in hypodermal cells 4 hours before ecdysis during all larval stages. The NAS-37 protein accumulates in the anterior cuticle and is shed in the cuticle after ecdysis. This pattern of protein accumulation places NAS-37 in the right place and at the right time to degrade the cuticle to facilitate ecdysis. The nas-37 gene has orthologs in other nematode species, including parasitic nematodes, and they undergo a similar shedding process. For example, Haemonchus contortus molts by digesting a ring of cuticle at the tip of the nose. Incubating Haemonchus larvae in extracted exsheathing fluids causes a refractile ring of digested cuticle to form at the tip of the nose. When Haemonchus cuticles are incubated with purified NAS-37, a similar refractile ring forms. NAS-37 degradation of the Haemonchus cuticle suggests that the metalloproteases and the cuticle substrates involved in exsheathment of parasitic nematodes are conserved in free-living nematodes. PMID- 15539495 TI - Antigenic diversity and immune evasion by malaria parasites. PMID- 15539496 TI - Expanding role of circulating adhesion molecules in assessing prognosis and treatment response in human immunodeficiency virus infection. PMID- 15539497 TI - Impact of immune plasticity on development of cellular memory responses to human immunodeficiency virus type 1. PMID- 15539498 TI - Comparison of enzyme-linked immunosorbent assay, Western blotting, microagglutination, indirect immunofluorescence assay, and flow cytometry for serological diagnosis of tularemia. AB - The serodiagnostic efficiencies of five different approaches to detecting antibodies (immunoglobulins G, A, and M) developed in clinically proven infections with Francisella tularensis have been assessed. Fifty serum samples from patients suffering from tularemia during an outbreak in Sweden were compared with samples from 50 healthy blood donors (controls) by using an enzyme-linked immunosorbent assay (ELISA), microagglutination (MA), Western blotting (WB), an indirect immunofluorescence assay (IIFA), and flow cytometry (FC). ELISA, WB, and FC were based on the use of preparations of lipopolysaccharides (LPS) of the live vaccine strain of Francisella tularensis subsp. holarctica (ATCC 29684) as a capture antigen. Whole methanol-fixed bacteria were used for IIFA and MA. Optimized protocols yielded a diagnostic sensitivity and specificity of 100% for WB, MA, and FC, 98% for ELISA, and 93% for IIFA. A total of 6,632 serum samples from individuals between the ages of 18 and 79 years, representatively recruited from all regions of Germany, were screened to estimate and confirm the positive predictive value (PVpos) of the ELISA. Serum samples from 15 (0.226%) individuals tested positive for F. tularensis-specific antibodies by ELISA and confirmatory WB. The resulting prevalence-dependent PVpos of 10.2% and specificity of 98.1% were consistent with our findings for tularemia patients and controls. We conclude that the combined usage of a screening ELISA and a confirmatory WB based on LPS as a common antigen, as well as the MA, is a suitable serodiagnostic tool, while the quality of the IIFA is hampered by subjective variations of the results. FC is a promising new approach that might be improved further in terms of multiplex analyses or high-throughput applications. PMID- 15539499 TI - Limited value of assays using detection of immunoglobulin G antibodies to the two recombinant dense granule antigens, GRA1 and GRA6 Nt of Toxoplasma gondii, for distinguishing between acute and chronic infections in pregnant women. AB - An enzyme-linked immunosorbent assay (ELISA) using two recombinant antigens of Toxoplasma gondii (GRA1 and GRA6 Nt) was developed in order to differentiate between pregnant women with a serological profile of recently acquired infection and those with chronic infection. Both proteins were expressed in Escherichia coli as glutathione S-transferase fusion proteins. Thirty-two serum samples from subjects who presented seroconversion within 3 months before sampling (group 1; acute profile), 46 serum samples from women who had a positive serology at least 1 year before sampling (group 2; chronic profile), and 100 serum samples from pregnant women who were not infected by T. gondii (group 3) were examined for immunoglobulin G (IgG) reactivity. For both antigens, the specificity reached 98%. In both groups of infected patients, the overall sensitivity scored was 60% for GRA1 and 83% for GRA6 Nt. In group 1, 34% of sera reacted with GRA1 whereas 84% of sera reacted with GRA6 Nt; in group 2, however, sensitivities were 78.2 and 82.6%, respectively. Combination of the readings obtained with both antigens yielded a sensitivity of 91%. A serological follow-up of 10 women who seroconverted during pregnancy displayed three different serological patterns: (i) a GRA profile paralleling the IgG curve, as detected by the commercial kit, (ii) a GRA1 profile, or (iii) GRA1 and GRA6 Nt profiles remaining negative for at least 8 weeks after the reference test gave positive results. Taken together, these results suggest that neither GRA1 nor GRA6 Nt is sensitive enough to be used routinely to differentiate between acute and chronic toxoplasmic infections. PMID- 15539500 TI - Use of antibodies in lymphocyte secretions for detection of subclinical tuberculosis infection in asymptomatic contacts. AB - We have previously demonstrated that Mycobacterium bovis BCG-specific immunoglobulin G antibodies in lymphocyte secretions (ALS) can be employed as a marker for active tuberculosis (TB). We aimed to determine whether the ALS method allows detection of subclinical TB infection in asymptomatic individuals. A prospective study of family contacts (FCs) of patients with active TB and healthy controls was performed. Thirteen of 42 FCs had high ALS responses, including 6 FCs who subsequently developed active TB. No correlation was observed between the tuberculin skin test and the ALS responses in the FCs (r = 0.1, P = 0.23). Among patients with active TB, BCG-specific ALS responses steadily declined from the time of diagnosis through 6 months following antimycobacterial chemotherapy (P = 0.001). The ALS assay enabled detection of infection in exposed symptom-free contacts, who are at greater risk for developing active TB. The method may also allow discrimination between effective treatment of active infection and suboptimal response to therapy. PMID- 15539502 TI - Antigenic determinants of alpha-like proteins of Streptococcus agalactiae. AB - The majority of group B streptococcus (GBS) isolates express one or more of a family of surface-anchored proteins that vary by strain and that form ladder-like patterns on Western blotting due to large repeat units. These proteins, which are important as GBS serotype markers and as inducers of protective antibodies, include the alpha C (Calpha) and R4 proteins and the recently described alpha like protein 2 (Alp2), encoded by alp2, and Alp3, encoded by alp3. In this study, we examined antigenic determinants possessed by Alp2 and Alp3 by testing of antibodies raised in rabbits, mainly by using enzyme-linked immunosorbent assays (ELISA) and an ELISA absorption test. The results showed that Alp2 and Alp3 shared an antigenic determinant, which may be a unique immunological marker of the Alp variants of GBS proteins. Alp2, in addition, possessed an antigenic determinant which showed specificity for Alp2 and a third determinant which showed serological cross-reactivity with Calpha. Alp3, in addition to the determinant common to Alp2 and Alp3, harbored an antigenic site which also was present in the R4 protein, whereas no Alp3-specific antigenic site was detected. These ELISA-based results were confirmed by Western blotting and a fluorescent antibody test. The results are consistent with highly complex antigenic structures of the alpha-like proteins in a fashion which is in agreement with the recently described structural mosaicism of the alp2 and alp3 genes. The results are expected to influence GBS serotyping, immunoprotection studies, and GBS vaccine developments. PMID- 15539501 TI - Diagnosis of norwalk virus infection by indirect enzyme immunoassay detection of salivary antibodies to recombinant norwalk virus antigen. AB - Simple diagnostic tests are needed for the detection of norovirus (NoV) outbreaks. Salivary antibody assays provide an attractive alternative to collecting and testing serum or stool samples. Antibodies to Norwalk virus (NV) in oral fluid samples were compared with NV antibodies in serum collected from 38 volunteers challenged with NV inoculum. Pre- and postchallenge (day 4, 8, 14, and 21) saliva and serum samples were examined by enzyme immunoassay (EIA) using recombinant NV antigen. Of 18 infected subjects (those who shed NV in stool or who demonstrated immunoglobulin G [IgG] seroconversion), 15 (83%) had > or =4 fold increases in NV-specific salivary IgA and 15 (83%) had > or =4-fold increases in NV-specific salivary IgG when prechallenge and postchallenge saliva samples were compared. When the results of the IgA and IgG assays were combined, all 18 infected subjects showed > or =4-fold increases in NV-specific salivary IgG or IgA postchallenge titers compared to their prechallenge titers. One of 19 uninfected subjects had a > or =4-fold increase in NV-specific salivary IgG. The sensitivity of the combined assay results was 100%, and the specificity was 95%. NV-specific salivary IgA titers peaked around 14 days postchallenge. NV-specific salivary IgG and serum IgG titers continued to rise through 21 days postchallenge. The application of this EIA to an elementary school outbreak indicated that 67% of the subjects with confirmed infections had >4-fold rises in anti-NoV IgA when an antigen in the same genetic cluster as the outbreak virus was used. This is the first documented mucosal antibody response to NoV in children. This EIA provides a useful approach for diagnosing NoV outbreaks. PMID- 15539503 TI - Overexpression of a novel lymphocyte population, positive for an intracellular CD14-like antigen, in patients positive for human immunodeficiency virus type 1. AB - CD14, originally recognized as a lipopolysaccharide (LPS) receptor, has recently been implicated in the process of T-cell suppression and apoptosis. Its soluble form has been shown to bind, in vitro, to human T cells, a process that may carry a negative signal onto these cells. We recently described a novel lymphocyte population in human peripheral blood, a population that expresses an intracellular CD14-like antigen. This novel T-cell population, composed mainly of CD8 cells and of very few CD4 cells, was found to be greatly enhanced in asymptomatic, untreated human immunodeficiency virus (HIV)-positive individuals. In the present study, we further characterized this cell population and found that it differed from other CD8 subpopulations associated with HIV infection such as CD8/CD38. In addition, we followed HIV patients under conditions of highly active antiretroviral therapy (HAART) and observed two groups of patients: patients in whom the CD14-like positive-testing T cells returned to normal within 1 to 3 months, and patients in whom it did not, in spite of a significant plasma HIV-RNA viral load decrease. Thus, this new CD14-like positive-testing lymphocyte population may represent an interesting and important component of the cellular events associated with HIV infection. On the basis of its modulation following HAART, we speculate that it may be used, in the future, as a drug-monitoring cellular marker in antiretroviral treatment. PMID- 15539504 TI - Establishment of diagnostic cutoff points for levels of serum antibodies to pertussis toxin, filamentous hemagglutinin, and fimbriae in adolescents and adults in the United States. AB - Numerous reports have documented that serologic methods are much more sensitive than culture for the diagnosis of pertussis in adolescents and adults. However, a standardized serologic test for pertussis is not routinely available to most clinicians, and the serologic test levels or cutoff points correlated with diseases have not been determined. The goal of the present study was to examine the distribution of immunoglobulin G (IgG) levels against three Bordetella pertussis antigens (pertussis toxin [PT], filamentous hemagglutinin [FHA], and fimbria types 2 and 3 [FIM]) and to determine population-based antibody levels for the purpose of establishing such diagnostic cutoff points. Enzyme-linked immunosorbent assays (ELISAs) were performed with sera from >6,000 U.S. residents aged 6 to 49 years who participated in the Third National Health and Nutrition Examination Survey. Mixture models were developed to identify hypothesized exposure groups and establish diagnostic cutoffs. Quantifiable (>20 ELISA units/ml [EU]) anti-FHA and anti-FIM IgG antibodies were common (65 and 62% of individuals, respectively), but quantifiable anti-PT IgG antibodies were less frequent (16%). Given the distributions of antibody levels, an anti-PT IgG level of > or =94 EU was proposed as the diagnostic cutoff point. Application of this cutoff point to culture-confirmed illness in a prior study investigating cough illness yielded a high diagnostic sensitivity (80%) and specificity (93%). A standardized ELISA for anti-PT IgG with a single serum sample appears to be useful for the identification of recent B. pertussis infection in adolescents and adults with cough illness. The PT cutoff point will be further evaluated in prospective studies of confirmed B. pertussis infection. PMID- 15539505 TI - Evaluation of multiplexed fluorescent microsphere immunoassay for detection of autoantibodies to nuclear antigens. AB - Antibodies to extractable nuclear antigens (ENA) are found in a variety of collagen vascular diseases. Determining the individual specificities of these antibodies is extremely useful in establishing the disease diagnosis and in some cases the prognosis. With a multiplexed fluorescent microsphere immunoassay, reactivity to five of the most diagnostically useful ENA was measured in 249 serum samples, including samples from 56 patients previously documented to have systemic lupus erythematosus (SLE). Results of the multiplexed assay were compared to results from established ENA enzyme-linked immunosorbent assays (ELISAs), and the agreement, sensitivity, and specificity, respectively, for the five ENA evaluated were as follows: SSA, 99.1, 100.0, and 98.8%; SSB, 98.6, 88.9, and 99.5%; Sm, 97.6, 95.8, and 97.9%; RNP, 97.2, 92.7, and 98.8%; Scl-70, 93.6, 50.0, and 99.0%. In the 56 confirmed SLE patients, the frequency of significant concentrations of autoantibodies with the multiplexed assay was 21.4% for SSA, 7.1% for SSB, 10.7% for Sm, 32.1% for RNP, and 0% for Scl-70. The new flow cytometric bead-based multiplexed assay showed excellent correlation with the well-established single-analyte ELISA methods for four of five the ENA markers investigated in this study. The most notable discrepancies between the two assays were for the Scl-70 antigen, which was most often resolved in favor of the multiplexed assay. Our studies show that the multiplexed microsphere-based immunoassay is a sensitive and specific method for the detection and semiquantitation of ENA antibodies in human sera. PMID- 15539506 TI - Monoclonal antibodies for specific detection of Encephalitozoon cuniculi. AB - Seven species-specific monoclonal antibodies (MAbs) were produced against Encephalitozoon cuniculi and characterized. The MAbs were immunoglobulin G, and when used for indirect microimmunofluorescence microscopy and Western immunoblot assays, they detected E. cuniculi originating from clinical samples. They did not cross-react with other Encephalitozoon species (E. intestinalis and E. hellem) or with a collection of gram-negative bacteria, yeast, and other parasites. The MAbs reacted primarily with 121-, 56-, 45-, 43-, and 41-kDa protein epitopes of E. cuniculi. These epitopes were demonstrated to be E. cuniculi species specific by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We developed MAbs to strains of E. cuniculi that can be used successfully to distinguish E. cuniculi from other microsporidial species in cultures established from clinical specimens. These MAbs may provide a specific, simple, rapid, and low-cost tool for the identification and diagnosis of infections due to microsporidia. PMID- 15539507 TI - Assignment of weight-based antibody units for 13 serotypes to a human antipneumococcal standard reference serum, lot 89-S(f). AB - Weight-based immunoglobulin G (IgG), IgM, IgA, and total Ig antibody assignments were made to human antipneumococcal standard reference serum lot 89-S, also known as lot 89-SF, for Streptococcus pneumoniae capsular polysaccharide (PnPs) serotypes 2, 6A, 8, 9N, 10A, 11A, 12F, 15B, 19A, 17F, 20, 22F, and 33F, as well as for C-polysaccharide (C-Ps), extending the standard's usefulness for pneumococcal vaccine evaluation beyond the original serotype 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F, and 23F assignments (S. A. Quataert, C. S. Kirch, L. J. Quackenbush Wiedl, D. C. Phipps, S. Strohmeyer, C. O. Cimino, J. Skuse, and D. V. Madore, Clin. Diagn. Lab. Immunol. 2:590-597, 1995). The additional 14 assignments were determined using an equivalence of absorbance method with an anti-PnPs serotype 6B reference enzyme-linked immunosorbent assay (EIA). To assure accuracy, anti-PnPs EIA for serotype 14 antibodies, a previously assigned serotype, was performed concurrently. This method assures consistency of the new microgram-per-microliter assignments with previous antiserotype assignments to lot 89-S. The sum of the experimentally derived isotype assignments for anti-PnPs serotypes in lot 89-S agrees well with the separately determined total Ig assignment for each serotype. The lot 89-S assignments for serotypes 1, 5, 6B, 14, 18C, 19F, and 23F were used for pneumococcal conjugate vaccine clinical trial evaluation and to generate data in efficacy trials where serological correlates for protection have been proposed. The assignment of antibody concentrations to additional pneumococcal serotypes in this reference reagent facilitates the consistent and accurate comparison of serum antibody concentrations across clinical trials. PMID- 15539508 TI - New latex bead agglutination assay for differential diagnosis of cattle infected with Mycobacterium bovis and Mycobacterium avium subsp. paratuberculosis. AB - Extensive studies have shown that the current assays used to identify cattle infected with Mycobacterium bovis or Mycobacterium avium subsp. paratuberculosis are not sufficiently sensitive and specific to detect all infected animals, especially animals recently infected with the pathogens. In the present report we show that these limitations might be overcome with a latex bead agglutination assay (LBAA). With the specific immunodominant epitope (ESAT6-p) of M. bovis, we developed an LBAA and enzyme immunoassay (EIA) for that purpose and compared them with the "gold standard" culture method and skin test for their efficacy in detecting bovine tuberculosis. When sera from control healthy cows (n = 10), M. avium subsp. paratuberculosis-positive cattle (naturally infected, n = 16; experimentally infected, n = 8), and M. bovis-positive cattle (naturally infected, n = 49;experimentally infected, n = 20) were applied to an EIA and an LBAA developed with ESAT6-p, the two tests showed similar sensitivity (97.1% by EIA, 95.7% by LBAA), high specificity (94.2% by EIA, 100% by LBAA), and a positive correlation (kappa value, 0.85; correlation rate, 93.2%; correlation coefficient, 0.64). Receiver operating characteristic analysis of EIA results and comparison with the culture method determined a suitable cutoff value at 0.469, with an area under the curve of 0.991 (95% confidence interval, 0.977 to 1.0). As LBAA didn't show any positive reactions with sera from uninfected control cows or M. avium subsp. paratuberculosis-infected cattle, which were confirmed to be free of M. bovis by culture or PCR, LBAA using the ESAT6-p can be a rapid and useful M. bovis diagnostic assay. The data suggest that rapid, sensitive, and specific assays can be developed with peptides containing immunodominant epitopes present in proteins uniquely expressed in M. bovis or M. avium subsp. paratuberculosis for differential diagnosis of cattle infected with M. bovis or M. avium subsp. paratuberculosis. PMID- 15539510 TI - Improved affinity of a human anti-Entamoeba histolytica Gal/GalNAc lectin Fab fragment by a single amino acid modification of the light chain. AB - We previously produced, in Escherichia coli, a human monoclonal antibody Fab fragment, CP33, specific for the galactose- and N-acetyl-D-galactosamine inhibitable lectin of Entamoeba histolytica. To prepare antibodies with a higher affinity to the lectin, recombination PCR was used to exchange Ser91 and Arg96 in the third complementarity-determining region of the light chain with other amino acids. The screening of 200 clones of each exchange by an indirect fluorescent antibody test showed that 14 clones for Ser91 and nine clones for Arg96 reacted strongly with E. histolytica trophozoites. Sequence analyses revealed that the substituted amino acids at Ser91 were Ala in five clones, Gly in three clones, Pro in two clones, and Val in two clones, while the amino acid at position 96 was substituted with Leu in three clones. The remaining eight clones exhibited no amino acid change at position 91 or 96. These mutant Fab fragments were purified and subjected to a surface plasmon resonance assay to measure the affinity of these proteins to the cysteine-rich domain of lectin. Pro or Gly substitution for Ser91 caused an increased affinity of the Fab, but substitution with Ala or Val did not. The replacement of Arg96 with Leu did not affect affinity. These results demonstrate that modification of antibody genes by recombination PCR is a useful method for affinity maturation and that amino acid substitution at position 91 yields Fabs with increased affinity for the lectin. PMID- 15539509 TI - Association of CD4+ CD25+ T cells with prevention of severe destructive arthritis in Borrelia burgdorferi-vaccinated and challenged gamma interferon-deficient mice treated with anti-interleukin-17 antibody. AB - CD4+ CD25+ T cells are a population of regulatory T cells responsible for active suppression of autoimmunity. Specifically, CD4+ CD25+ T cells have been shown to prevent insulin-dependent diabetes mellitus, inflammatory bowel disease, and pancreatitis. Here, we present evidence that CD4+ CD25+ T cells also play a major role in controlling the severity of arthritis detected in Borrelia burgdorferi vaccinated gamma interferon-deficient (IFN-gamma degrees ) C57BL/6 mice challenged with the Lyme spirochete. When B. burgdorferi-vaccinated and challenged IFN-gamma degrees mice were treated with anti-interleukin-17 (IL-17) antibody, the number of CD4+ CD25+ T cells increased in the local lymph nodes. Furthermore, histopathologic examination showed the mice to be free of destructive arthritis. When these anti-IL-17-treated B. burgdorferi-vaccinated and challenged mice were also administered anti-CD25 antibody, the number of CD4+ CD25+ T cells in the local lymph nodes decreased. More importantly, severe destructive arthropathy was induced. In addition, delayed administration of anti CD25 antibody decreased the severity of the arthritis. These results suggest that CD4+ CD25+ T cells are involved in regulation of a severe destructive arthritis induced with an experimental model of vaccination and challenge with B. burgdorferi. PMID- 15539511 TI - Evaluation of an in vitro assay for gamma interferon production in response to Mycobacterium tuberculosis infections. AB - The tuberculin skin test (TST) is the "gold standard" for detecting infection with Mycobacterium tuberculosis. We compared the TST using purified protein derivative to the QuantiFERON-TB test (QFT). Two groups were examined. Group 1 individuals (n = 66) (low risk) were at low risk for exposure to M. tuberculosis and were not Mycobacterium bovis BCG vaccinated. Group 2 (n = 29) include individuals who were likely to have been exposed to a high prevalence of M. tuberculosis infections and were BCG vaccinated. Group 1 individuals were given a TST. Group 2 individuals were not given a TST because of possible adverse reactions. A 10- to 15-mm indurated area 48 h after TST was considered positive. A positive QFT result was defined as a significant gamma interferon response to M. tuberculosis antigen, Mycobacterium avium antigen, and a nonspecific mitogen stimulus and no response in the negative control. In group 1, 60 of 66 individuals (90.9%) were negative by both methods, and 1 person was positive by both methods. There was one QFT-negative, TST-positive case, one QFT-positive, TST-negative case, and three conditional QFT-positive, TST-negative cases. In group 2, 12 of 29 (41.4%) were positive by QFT and considered likely to be TST positive because of prior BCG vaccination. QFT testing in our low-risk group resulted in an agreement of 96.8%, a sensitivity of 50%, and a specificity of 98.4% compared with TST results. QFT testing with TST in low-risk groups can aid in the detection of latent M. tuberculosis infections. PMID- 15539512 TI - Recombinant Helicobacter bilis protein P167 for mouse serodiagnosis in a multiplex microbead assay. AB - Infection of mice with Helicobacter bilis is widespread in research and commercial mouse colonies. Therefore, sensitive, specific, and high-throughput assays are needed for rapid and accurate testing of mice in large numbers. This report describes a novel multiplex assay, based on fluorescent microbeads, for serodetection of H. bilis infection. The assay requires only a few microliters of serum to perform and is amenable to a high-throughput format. Individual microbead sets were conjugated to purified, H. bilis-specific, recombinant proteins P167C and P167D and bacterial membrane extracts from H. bilis and Helicobacter hepaticus. For detecting H. bilis infection in the microbead multiplex assay, P167C and P167D provided significantly higher sensitivities (94 and 100%, respectively) and specificities (100 and 95%, respectively) than membrane extract (78% sensitivity and 65% specificity). Microbead multiplex assay results were validated by enzyme-linked immunosorbent assay. Purified recombinant proteins showed low batch-to-batch variation; this feature allows for ease of quality control, assay robustness, and affordability. Thus, recombinant antigens are highly suitable in the multiplex microbead assay format for serodetection of H. bilis infection. PMID- 15539513 TI - Effects of prior polysaccharide vaccination on magnitude, duration, and quality of immune responses to and safety profile of a meningococcal serogroup C tetanus toxoid conjugate vaccination in adults. AB - Extensive use of meningococcal AC polysaccharide (MACP) vaccines has raised concerns about induction of immunologic hyporesponsiveness to C polysaccharide. We investigated the immunogenicity and safety of a meningococcal C-tetanus conjugate (MCC-TT) vaccine in naive adults and prior MACP vaccinees. Laboratory staff (n = 113) were recruited; 73 were naive to meningococcal vaccination, and 40 had previously received > or =1 dose of MACP vaccine. Blood was taken prior to MCC-TT vaccination and 1 week, 1 month, and 6 months later. At each time point, proportions of subjects with serum bactericidal antibody (SBA) titers of > or =8 or > or =128 were similar (P > 0.46); >94% of subjects achieved titers of > or =128 at 1 month. However, the geometric mean titer (GMT) of SBA at 1 month was higher in the naive (1,757; 95% confidence interval [95% CI], 1,102 to 2,803) than in the previously vaccinated (662; 95% CI, 363 to 1,207) group (P = 0.02), and similarly at 6 months (P < 0.001). Conversely, geometric mean concentrations (GMCs) of serogroup C-specific immunoglobulin G (IgG) were significantly higher in the previously vaccinated group pre-MCC-TT and at 1 week; the groups were similar at 1 month, and there was some evidence that the GMC for the previously vaccinated group was higher at 6 months. Qualitative differences in antibodies between groups were demonstrated by using the SBA/IgG ratio, though avidity measures were similar for the two groups throughout the study. MCC-TT was well tolerated, with similar safety profiles in the two groups. Pain in the arm and headache were the most frequently reported events following vaccination. The study shows that MCC-TT is safe and immunogenic in naive and previously MACP vaccinated adults, though the magnitude and persistence of postvaccination SBA responses in the latter group were lower. PMID- 15539514 TI - Establishing phenotypic features associated with morbidity in human T-cell lymphotropic virus type 1 infection. AB - The human T-cell lymphotropic virus type 1 (HTLV-1) is the causative agent of HTLV-1-associated myelopathy/tropical spastic paraparesis (HT). Although it is widely believed that virus infection and host immune response are involved in the pathogenic mechanisms, the role of the immune system in the development and/or maintenance of HT remains unknown. We performed an analysis of the peripheral blood leukocyte phenotype for two different subcohorts of HTLV-1-infected individuals to verify the existence of similar immunological alterations, possible laboratory markers for HT. The leukocyte population balance, the activation status of the T lymphocytes, and the cellular migratory potential of T lymphocytes, monocytes, and neutrophils were evaluated in the peripheral blood of HTLV-1-infected individuals classified as asymptomatic individuals, oligosymptomatic individuals, and individuals with HT. Data analysis demonstrated that a decreased percentage of B cells, resulting in an increased T cell/B cell ratio and an increase in the CD8+ HLA-DR+ T lymphocytes, exclusively in the HT group could be identified in both subcohorts, suggesting its possible use as a potential immunological marker for HT for use in the laboratory. Moreover, analysis of likelihood ratios showed that if an HTLV-1-infected individual demonstrated B-cell percentages lower than 7.0%, a T cell/B cell ratio higher than 11, or a percentage of CD8+ HLA-DR+ T lymphocytes higher than 70.0%, this individual would have, respectively, a 12-, 13-, or 22-times-greater chance of belonging to the HT group. Based on these data, we propose that the T cell/B cell ratios and percentages of circulating B cells and activated CD8+ T lymphocytes in HTLV-1-infected patients are important immunological indicators which could help clinicians monitor HTLV-1 infection and differentiate the HT group from the asymptomatic and oligosymptomatic groups. PMID- 15539515 TI - Regulation of fungal infection by a combination of amphotericin B and peptide 2, a lactoferrin peptide that activates neutrophils. AB - To establish a novel strategy for the control of fungal infection, we examined the antifungal and neutrophil-activating activities of antimicrobial peptides. The duration of survival of 50% of mice injected with a lethal dose of Candida albicans (5 x 10(8) cells) or Aspergillus fumigatus (1 x 10(8) cells) was prolonged 3 to 5 days by the injection of 10 microg of peptide 2 (a lactoferrin peptide) and 10 microg of alpha-defensin 1 for five consecutive days and was prolonged 5 to 13 days by the injection of 0.1 microg of granulocyte-monocyte colony-stimulating factor (GM-CSF) and 0.5 microg of amphotericin B. When mice received a combined injection of peptide 2 (10 microg/day) with amphotericin B (0.5 microg/day) for 5 days after the lethal fungal inoculation, their survival was greatly prolonged and some mice continued to live for more than 5 weeks, although the effective doses of peptide 2 for 50 and 100% suppression of Candida or Aspergillus colony formation were about one-third and one-half those of amphotericin B, respectively. In vitro, peptide 2 as well as GM-CSF increased the Candida and Aspergillus killing activities of neutrophils, but peptides such as alpha-defensin 1, beta-defensin 2, and histatin 5 did not upregulate the killing activity. GM-CSF together with peptide 2 but not other peptides enhanced the production of superoxide (O2-) by neutrophils. The upregulation by peptide 2 was confirmed by the activation of the O2- -generating pathway, i.e., activation of large-molecule guanine binding protein, phosphatidyl-inositol 3-kinase, protein kinase C, and p47phox as well as p67phox. In conclusion, different from natural antimicrobial peptides, peptide 2 has a potent neutrophil-activating effect which could be advantageous for its clinical use in combination with antifungal drugs. PMID- 15539516 TI - Serological method using recombinant S2 protein to differentiate equine infectious anemia virus (EIAV)-infected and EIAV-vaccinated horses. AB - We recently reported a highly protective attenuated live virus vaccine for equine infectious anemia virus (EIAV) based on a proviral construct (EIAVUKDeltaS2) with a genetically engineered mutation in the viral S2 gene that eliminates expression of this accessory protein. While the EIAVUKDeltaS2 vaccine provides protection from detectable infection by experimental challenge with highly virulent virus, the potential for commercial application of this vaccine is complicated by the fact that horses inoculated with the EIAVUKDeltaS2 vaccine strain become seropositive in various reference diagnostic assays based on detection of antibodies to virion core or envelope proteins. To address this issue, we describe here the development and optimization of a new serologic EIAV diagnostic enzyme-linked immunosorbent assay (ELISA) to detect serum antibodies to the EIAV S2 protein that are produced in infected horses but not in horses inoculated with the EIAVUKDeltaS2 vaccine virus. The test S2 protein antigen was developed using the S2 gene sequence from the EIAVUK strain of virus and a series of modifications to facilitate production and purification of the diagnostic antigen, designated HS2G. Using this HS2G as antigen, we describe the development of an affinity ELISA that provides a sensitive and specific detection of S2 specific serum antibodies in experimentally and field-infected horses (22 of 24), without detectable reactivity with immune serum from uninfected (12 of 12) or vaccinated (29 of 29) horses. These data indicate that the S2-based diagnostic ELISA has the potential to accurately differentiate horses infected with EIAV from horses inoculated with an attenuated EIAV vaccine strain with a mutant S2 gene. PMID- 15539517 TI - Evaluation of a diagnostic algorithm using immunoglobulin M enzyme-linked immunosorbent assay to differentiate human West Nile Virus and St. Louis Encephalitis virus infections during the 2002 West Nile Virus epidemic in the United States. AB - A diagnostic algorithm was developed to differentiate between human infections of West Nile virus (WNV) and St. Louis encephalitis virus (SLEV) using positive-to negative (P/N) ratios derived from the immunoglobulin M capture enzyme-linked immunosorbent assay (MAC-ELISA). To validate this algorithm, we tested 1,418 serum and cerebrospinal fluid (CSF) samples from confirmed WNV and SLEV infections collected during the WNV epidemic of 2002 in the United States. WNV P/N-to-SLEV P/N ratios (W/S ratios) were calculated and used to identify the infecting virus. These results were compared to results from the plaque reduction neutralization test (PRNT), which is currently the standard assay used to discriminate between closely related flavivirus infections. If the W/S ratio was > or =1, the predictive value positive (PNP) for WNV was 97.8%, where 95% of flavivirus cases were due to WNV infection and only 3.7% of specimens would require PRNT to differentiate WNV from SLEV infection. Use of the W/S ratio as part of the testing algorithm to interpret MAC-ELISA results generates reportable probable cases quickly, alleviating the need for PRNT in most instances. PMID- 15539518 TI - Infection of immunodeficient horses with Sarcocystis neurona does not result in neurologic disease. AB - Equine protozoal myeloencephalitis is a progressive neurologic disease of horses most commonly caused by infection with the apicomplexan parasite Sarcocystis neurona. Factors affecting neuroinvasion and neurovirulence have not been determined. We investigated the pathogenesis of infection with S. neurona in horses with severe combined immune deficiency (SCID). Two immunocompetent (IC) Arabian horses and two Arabian horses with SCID were infected orally with 5 x 10(5) sporocysts of S. neurona. Four IC horses and one SCID horse were infected intravenously (i.v.) with 5 x 10(8) merozoites of the WSU-1 isolate of S. neurona. Despite prolonged parasitemia and persistent infection of visceral tissues (skeletal muscle, cardiac muscle, lung, liver, and spleen) as demonstrated by PCR and culture, SCID horses did not develop neurologic signs after oral or i.v. infection. S. neurona was undetectable in the neuronal tissues of SCID horses by either PCR, immunohistochemistry, or culture. In contrast, although parasitemia was undetectable in orally infected IC horses and of only short duration in i.v. infected IC horses, four of six IC horses developed neurologic signs. S. neurona was detectable by PCR and/or culture of neural tissue but not visceral tissue of IC horses with neurologic disease. Infected SCID horses are unable to clear S. neurona from visceral tissues, but the infection does not result in neurologic signs; in contrast, IC horses rapidly control parasitemia and infection of visceral tissues but frequently experience neuroinvasion and exhibit clinical signs of neurologic disease. PMID- 15539519 TI - Bikunin inhibits lipopolysaccharide-induced tumor necrosis factor alpha induction in macrophages. AB - Bikunin, a Kunitz-type protease inhibitor, exhibits anti-inflammatory activity in protection against cancer and inflammation. To investigate the molecular mechanism of this inhibition, we analyzed the effect of bikunin on tumor necrosis factor alpha (TNF-alpha) production in human peripheral mononuclear cells stimulated by lipopolysaccharide (LPS), an inflammatory inducer. Here, we show the following results. (i) LPS induced TNF-alpha expression in time- and dose dependent manners through phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 mitogen activated protein kinase pathways. (ii) Bikunin inhibits LPS-induced up regulation of TNF-alpha protein expression in a dose-dependent manner, reaching 60% inhibition at the highest doses of bikunin tested (5.0 microM). (iii) Inhibition by bikunin of TNF-alpha induction correlates with the suppressive capacity of ERK1/2, JNK, and p38 signaling pathways, implicating repressions of at least three different signals in the inhibition. (iv) Bikunin blocks the induction of TNF-alpha target molecules interleukin-1beta (IL-1beta) and IL-6 proteins. (v) Bikunin is functional in vivo, and this glycoprotein blocks systemic TNF-alpha release in mice challenged with LPS. (vi) Finally, bikunin can prevent LPS-induced lethality. In conclusion, bikunin significantly inhibits LPS induced TNF-alpha production, suggesting a mechanism of anti-inflammation by bikunin through control of cytokine induction during inflammation. Bikunin might be a candidate for the treatment of inflammation, including septic shock. PMID- 15539520 TI - Use of viral lysate antigen combined with recombinant protein in Western immunoblot assay as confirmatory test for serodiagnosis of severe acute respiratory syndrome. AB - A Western immunoblot assay for confirmatory serodiagnosis of severe acute respiratory syndrome (SARS) was developed utilizing viral lysate antigens combined with a recombinant nucleocapsid protein, GST-N (glutathione S transferase-nucleocapsid) of the SARS coronavirus (SARS-CoV). The viral lysate antigens were separated by electrophoresis and transblotted onto nitrocellulose membranes. The resultant membrane was subsequently added with the GST-N recombinant protein at a specific location. The positions of bands corresponding to some of the structural proteins immobilized on the membrane were then located and verified with mouse or rabbit antisera specific to the respective proteins. The Western immunoblot assay was able to detect antibodies to SARS-CoV in all 40 serum specimens from SARS patients and differentiate the SARS-positive samples from those of the healthy donor or non-SARS patient controls (150 samples) when set criteria were followed. In addition, when the immunoblot was used to test samples considered falsely positive by an in-house-developed SARS-specific enzyme linked immunosorbent assay, band patterns different from those with samples from SARS patients were obtained. PMID- 15539521 TI - Structural and immunological characteristics of chronically inflamed adenotonsillar tissue in childhood. AB - Recurrent or chronic adenotonsillar infections mainly affect children and frequently involve otherwise healthy subjects. Therefore, having excluded systemic immunological deficiencies, this disease may be due to a local dysfunction of the epithelial structures at either the rhino or oropharyngeal level. The aim of the present investigation was to analyze structural and immunological aspects of tonsils and adenoids in subjects who underwent adenotonsillectomy because of recurrent inflammatory episodes with fever. Histological studies and analyses of the cytokine patterns were carried out in palatine tonsils and adenoid samples from 105 patients who underwent adenoidectomy and bilateral extracapsular tonsillectomy for chronic inflammatory hypertrophy of these organs; 46 of the 105 cases examined presented hyperkeratosis of the crypt epithelium; in the remaining 59, the epithelium was hyperplastic with no signs of keratosis. Scanning electron microscopy revealed a continuous epithelial surface of polygon-shaped flattened cells with fissures towards the cryptic depressions. Titration of interleukin-1beta and tumor necrosis factor alpha in serum and tissues demonstrated higher concentrations in the adenotonsillar specimens, whereas the rise in interleukin-6 was more modest. PMID- 15539522 TI - Characterization of antibodies to capsular polysaccharide antigens of Haemophilus influenzae type b and Streptococcus pneumoniae in human immune globulin intravenous preparations. AB - The most common infections in primary immune deficiency disease (PIDD) patients involve encapsulated bacteria, mainly Haemophilus influenzae type b (Hib) and Streptococcus pneumoniae (pneumococcus). Thus, it is important to know the titers of Hib- and pneumococcus-specific antibodies that are present in immune globulin (Ig) intravenous (IGIV) preparations used to treat PIDD. In this study, seven IGIV preparations were tested by enzyme-linked immunosorbent assay and opsonophagocytic activity for antibody titers to the capsular polysaccharides of Hib and five pneumococcal serotypes. Differences in Hib- and pneumococcus specific antibody titer were observed among various IGIV preparations, with some products having higher- or lower-than-average titers. Opsonic activity also varied among preparations. As expected, IgG2 was the most active subclass of both binding and opsonic activity except against pneumococcal serotype 6B where IgG3 was the most active. This study determines antibody titers against capsular polysaccharides of Hib and pneumococcus in seven IGIV products that have been shown to be effective in reducing infections in PIDD patients. As donor antibody levels and manufacturing methods continue to change, it may prove useful from a regulatory point of view to reassess IGIV products periodically, to ensure that products maintain antibody levels that are important for the health of IGIV recipients. PMID- 15539523 TI - Utility of Schistosoma bovis adult worm antigens for diagnosis of human schistosomiasis by enzyme-linked immunosorbent assay and electroimmunotransfer blot techniques. AB - Immunodiagnostic methods based on the detection of antibodies continue to be the most effective and practical methods for the diagnosis of imported schistosomiasis. Schistosoma bovis is a species whose final natural hosts are bovines, ovines, caprines, and small wild ruminants. Different studies have demonstrated the analogies existing between S. bovis and other Schistosoma species which affect humans. The objective of this work was to evaluate the utility of S. bovis adult worm antigens (AWA) for the diagnosis of imported human schistosomiasis by enzyme-linked immunosorbent assay (ELISA) and electroimmunotransfer blotting (EITB) techniques. By detecting eggs, the ELISA for S. bovis AWA was able to definitively detect imported cases with a sensitivity of 94%. The specificity of the ELISA for S. bovis AWA was 97%. There were no differences between the results of the S. bovis AWA ELISA for patients infected with Schistosoma mansoni and those infected with Schistosoma haematobium. The EITB technique showed bands of 85, 37, and 20 kDa, which are characteristic of infections with Schistosoma spp. Specific bands to indicate infection by different species of Schistosoma have not been detected. The combined use of the ELISA for S. bovis AWA and EITB increased the global sensitivity of the study to 97%. Our findings suggest that the ELISA for S. bovis AWA is a useful test for the immunodiagnosis of imported schistosomiasis and that EITB for detecting S. bovis AWA permits the confirmation of diagnosis when the ELISA for S. bovis AWA is positive. PMID- 15539525 TI - Specific immunoglobulin g antibody detected in umbilical blood and amniotic fluid from a pregnant woman infected by the coronavirus associated with severe acute respiratory syndrome. AB - Specific immunoglobulin G antibody for severe acute respiratory syndrome (SARS) coronavirus was detected in maternal blood, umbilical blood, and amniotic fluid from a pregnant SARS patient. Potential protection of fetus from infection was suggested. PMID- 15539524 TI - Induction of a humoral immune response following an Escherichia coli O157:H7 infection with an immunomodulatory peptidic fraction derived from Lactobacillus helveticus-fermented milk. AB - Numerous beneficial effects have been attributed to probiotic lactic acid bacteria (LAB), such as the stimulation of the immune system, the prevention of enteric infections by enteropathogens, and the regression of immunodependent tumors. It has been shown that biologically active metabolites released during fermentation, in particular biopeptides, could act as immunomodulatory agents. However, no studies have been conducted to evaluate the implication of these bioactive peptides in the induction of a protective immune response against enteric infections. The present study aimed to evaluate the possible immunomodulatory and anti-infectious effects of a peptidic fraction released in milk fermented by Lactobacillus helveticus. The immune response in the mucosa associated lymphoid tissue was monitored following an administration of the potentially bioactive peptidic fraction. The total immunoglobulin A (IgA) immune response was evaluated after an Escherichia coli O157:H7 infection in a BALB/c murine model. Immunohistochemical and enzyme-linked immunosorbent assays revealed an increase in the number of IgA-secreting B lymphocytes in the intestinal lamina propria and an enhanced total secretory and systemic IgA response. Cytokine profiling also revealed stimulation of a Th2 response in mice fed the peptidic fraction, whereas infected controls demonstrated a proinflammatory Th1 response. These results indicate that bioactive peptides released during fermentation by LAB could contribute to the known immunomodulatory effects of probiotic bacteria. PMID- 15539526 TI - Age-dependent accuracy of Helicobacter pylori antibody assays for adults, with special emphasis on atrophic gastritis. AB - The accuracy of Helicobacter pylori antibody assays for 561 consecutive adult outpatients who had undergone gastroscopy was studied. The sensitivity of an immunoglobulin G test was 99 to 100% for all age groups, but the specificity declined by age group, from 99% for those aged 15 to 49 years to 75% for those aged > or =65 years. The exclusion of false-positive results for patients with atrophic gastritis improved the specificity to 93 to 97% for the older age groups. PMID- 15539527 TI - Expression of tocopherol-associated protein in mast cells. AB - Tocopherol-associated protein (TAP) was expressed in mouse mast cells. TAP was predominantly localized in the cytoplasm, and the subcellular localization was not changed by alpha-tocopherol. The results suggest that the physiological role of TAP in mast cells is not regulation of tocopherol function but an as-yet unidentified activity. PMID- 15539528 TI - Biased immunoglobulin G (IgG) subclass production in a case of hyper-IgM syndrome. AB - Hyper-immunoglobulin M (IgM) syndrome (HIGM) is a rare heterogeneous primary immune deficiency. We describe a patient with HIGM characterized by skewed production of serum IgG subclasses and normal somatic hypermutation. This case may represent a subgroup of HIGM type 4 that is characterized by a biased switching to the V-region proximal constant regions. PMID- 15539530 TI - Editorial: finding a research home. PMID- 15539529 TI - Lack of association between Toll-like receptor 2 polymorphisms and susceptibility to severe disease caused by Staphylococcus aureus. AB - To investigate a putative link between genetically determined variations in Toll like receptor 2 (TLR2) and the occurrence of severe Staphylococcus aureus infection, the functional Arg753Gln single-nucleotide polymorphism and the GT repeat microsatellite in the TLR2 gene were examined in a large case-control study. No associations with disease or mortality attributable to these features were found. PMID- 15539532 TI - Nursing care and context in Theresienstadt. AB - Over the past 5 years, nurses'roles as caregivers in ghettos and concentration camps during the Third Reich increasingly have become the subject of interest. Historians have focused particularly on the ethical dilemmas of caregiving practices that often deviated from normal care standards and situations. This article examines nurses and their work in one concentration camp, Theresienstadt, to demonstrate that even under the unique and horrible conditions of camp existence, contextual differences between camps provided nurses with different difficulties and opportunities. Although an extreme example, this article demonstrates the critical link between care and context as nurses practice across various settings and situations. PMID- 15539531 TI - Postoperative pain communication skills for older adults. AB - This study tested the effect of a preoperative pain communication intervention on older adults' ability to obtain pain relief after a total knee arthroplasty. A posttest-only experimental design was used to compare older adults randomly assigned to (a) view a pain management and pain communication film, (b) view the pain management film only, or (c) receive standard care only. Initial method adjustments decreased potential error in the study. Adjustments included testing only total knee arthroplasty patients receiving standard physical therapy and omitting unreliable measures from the analyses. Recruitment of the standard care group was halted when differences emerged between the remaining groups. Older adults in the communication group reported significantly less sensory pain on postoperative Day 1 than older adults in the pain management only group. Teaching older adults both pain communication skills and pain management information before surgery might result in greater pain relief during the early postoperative period. PMID- 15539533 TI - Cancer risk-reduction behaviors of breast cancer survivors. AB - Using secondary data analysis, the aim was to determine if postmenopausal women, who have survived breast cancer, have adopted healthy nutritional and physical activity behaviors recommended in the American Cancer Society guidelines as cancer risk-reduction strategies, and in guidelines for prevention of other chronic diseases or for improving general health. From their personal health history, women who have survived breast cancer would be likely candidates to adopt healthy behaviors recommended as cancer risk-reduction strategies or for prevention of other chronic diseases. A secondary aim was to determine the perceived general health and affective state of these women. These breast cancer survivors had a high perception of their general health, a positive affective state, and have adopted some healthy lifestyle behaviors, but they are not fully adhering to the ACS nutrition and physical activity guidelines or other health related guidelines for cancer risk reduction or prevention of other chronic diseases. PMID- 15539534 TI - Searching the literature by design. AB - Current nursing knowledge has been generated by studies whose designs range from exploratory to experimental. Searching the literature for a study illustrating a particular design can be a perplexing process. This article explains strategies that are useful in identifying research studies according to the designs used in their methods. The suggested search strategies assist in identifying the most relevant search results and can save time. Outcomes of such a search can be instructional about the implementation of a design in its classic form or some variation. They can provide practical illustrations of the advantages and limitations of a design when applied in a particular area of interest. PMID- 15539535 TI - Effect of environment and research participant characteristics on data quality. AB - The purpose of this study, a component of a randomized clinical trial, was to assess the influence of the emergency department environment and participant characteristics on the accuracy of self-reported health care utilization. Interviews of 612 seniors aged 65 to 93 were conducted in two emergency departments. The research assistant, upon completion of each interview, rated characteristics of the emergency department and compared participants' self reports of emergency department use and hospitalization during the previous 4 weeks with data from hospital records: 3.6% overreported and 2.2% underreported visits to the emergency department. Regarding hospitalizations, 2.6% overreported and 1.2% underreported. Discrepancies were associated with male gender, cognitive deficits, and risk status. Inconsistencies were not related to any of the environmental variables. These findings suggest that seniors without cognitive decline report reliable data even in a potentially challenging environment. PMID- 15539536 TI - How to develop a budget for a research proposal. AB - Novice investigators may be intimidated by the task of proposal budget preparation. Often a basic understanding of the mechanics of budgeting, paired with a good working relationship with the institution's sponsored programs office, can alleviate much of the stress investigators encounter in developing budgets. Careful attention to the detailed inclusive costs of conducting the proposed study combined with awareness of university and agency budget requirements is essential. This article describes strategies for developing a budget for a research proposal. PMID- 15539537 TI - Clinical decision rules as tools for evidence-based nursing. AB - Nursing practice is fraught with uncertainty and our patients do not always respond predictably to our interventions. Properly developed and tested Clinical Decision Rules (CDRs), a special type of decision support tool, help organize research evidence into standardized patient assessments and treatments, thereby increasing the probability of attaining the desired outcome and reducing uncertainty in practice. The purpose of this article is to examine CDRs as one kind of decision support tool that can be used to facilitate nursing decision making and evidence-based practice (EBP). CDRs differ from both treatment algorithms and clinical pathways in the ways they are developed and in their scope of applicability. CDRs are developed using strict methodological standards, with the goal that they function well when used by a variety of care providers, with various patient populations, and in different patient care settings. PMID- 15539541 TI - Toward a unified system of accreditation for professional preparation in health education: final report of the National Task Force on Accreditation in Health Education. AB - During the past 40 years, health education has taken significant steps toward improving quality assurance in professional preparation through individual certification and program approval and accreditation. Although the profession has begun to embrace individual certification, program accreditation in health education has been neither uniformly available nor universally accepted by institutions of higher education. To further strengthen professional preparation in health education, the Society for Public Health Education (SOPHE) and the American Association for Health Education (AAHE) established the National Task Force on Accreditation in Health Education in 2001. The 3-year Task Force was charged with developing a detailed plan for a coordinated accreditation system for undergraduate and graduate programs in health education. This article summarizes the Task Force's findings and recommendations, which have been approved by the SOPHE and AAHE boards, and, if implemented, promise to lay the foundation for the highest quality professional preparation and practice in health education. PMID- 15539542 TI - Ethical challenges for the "outside" researcher in community-based participatory research. AB - Although community-based participatory research (CBPR) shares many of the core values of health education and related fields, the outside researcher embracing this approach to inquiry frequently is confronted with thorny ethical challenges. Following a brief review of the conceptual and historical roots of CBPR, Kelly's ecological principles for community-based research and Jones's three-tiered framework for understanding racism are introduced as useful frameworks for helping explore several key challenges. These are (a) achieving a true "community driven" agenda; (b) insider-outsider tensions; (c) real and perceived racism; (d) the limitations of "participation"; and (e) issues involving the sharing, ownership, and use of findings for action. Case studies are used in an initial exploration of these topics. Green et al.'s guidelines for appraising CBPR projects then are highlighted as an important tool for helping CBPR partners better address the challenging ethical issues often inherent in this approach. PMID- 15539543 TI - A meta-analysis of adolescent psychosocial smoking prevention programs published between 1978 and 1997 in the United States. AB - Psychosocial smoking prevention studies have shown inconsistent results and theory-driven programs have been related to program success. This meta-analysis was used as a judgment tool for resolving these issues by estimating average program effects and investigating the relative efficacy of program types. The present study examined 65 adolescent psychosocial smoking prevention programs (1978 to 1997) among students in Grades 6 to 12 in the United States. Three program modalities (social influence, cognitive behavior, life skill) and two program settings (exclusively school based, school-community-incorporated) were identified as major a priori classifications. Knowledge had the highest effect sizes (.53) at short-term (< or = 1 year) but rapidly decreased (.19) at long term (> 1 year). Behavioral effect was the most meaningful, being persistent over a 3-year period (.19 at < or = 1 year; .18 at 1 to 3 years). Adolescent smoking reduction rates were increased by using either cognitive behavior or life skills program modalities, and/or a school-community-incorporated program setting. PMID- 15539544 TI - Increasing readers' comprehension of health education brochures: a qualitative study into how professional writers make texts coherent. AB - The aim of this study was to gain insight into the extent to which health education text writers apply writing principles derived from cognitive psychological theory. Seventeen professional text writers of health education materials participated in a qualitative study, consisting of a rewriting task combined with a think-aloud procedure and a semistructured interview. The verbal data were explored carefully in light of seven text coherence principles that have proven effective in cognitive psychological research to increase text comprehension. Findings indicate text writers vary in their ideas and use of coherence principles to make a text comprehensible. It is argued that the health education profession can benefit greatly from knowledge about cognitive text processing and cognitive coherence principles for realizing effective comprehension of written health education messages. PMID- 15539545 TI - Using the extended parallel process model to prevent noise-induced hearing loss among coal miners in Appalachia. AB - Occupational noise-induced hearing loss is the second most self-reported occupational illness or injury in the United States. Among coal miners, more than 90% of the population reports a hearing deficit by age 55. In this formative evaluation, focus groups were conducted with coal miners in Appalachia to ascertain whether miners perceive hearing loss as a major health risk and if so, what would motivate the consistent wearing of hearing protection devices (HPDs). The theoretical framework of the Extended Parallel Process Model was used to identify the miners' knowledge, attitudes, beliefs, and current behaviors regarding hearing protection. Focus group participants had strong perceived severity and varying levels of perceived susceptibility to hearing loss. Various barriers significantly reduced the self-efficacy and the response efficacy of using hearing protection. PMID- 15539546 TI - Assessing intervention effects in a school-based nutrition intervention trial: which analytic model is most powerful? AB - This article compares four mixed-model analyses valid for group-randomized trials (GRTs) involving a nested cohort design with a single pretest and posttest. This study makes estimates of intraclass correlations (ICCs) available to investigators planning GRTs addressing dietary outcomes. It also provides formulae demonstrating the potential benefits to the standard error of the intervention effect (sigma(delta)) from adjustments for both fixed and time varying covariates and correlations over time. These estimates will allow other researchers using these variables to plan their studies by estimating a priori detectable differences and sample size requirements for any of the four analytic options. These methods are demonstrated using data from the Teens Eating for Energy and Nutrition at School study. Mixed-model analyses of covariance proved to be the most powerful analysis in that data set. The formulae may be applied to any dependent variable in any GRT given corresponding information for those variables on the parameters that define the formulae. PMID- 15539547 TI - A qualitative study of substance use and sexual behavior among 18- to 29-year-old men while incarcerated in the United States. AB - The article describes men's perceptions of and experience with substance use and sexual behavior during incarceration. Grounded theory content analyses were performed on qualitative interviews conducted with 80 men, aged 18 to 29, in four U.S. states. Participants believed that drugs were easily available in prison. Half reported using substances, primarily marijuana or alcohol, while incarcerated. Key themes included the role of correctional personnel in the flow of substances in prison and the economic significance of substance trafficking. With regard to sexual behavior, most men acknowledged that it occurred but were hesitant to talk in-depth about it. There was a strong belief in "don't look, don't tell," and sex in prison was often associated with homosexual behavior or identity. Sex during incarceration was reported by 12 men, mostly with female partners. Participants were pessimistic about HIV/STD/hepatitis prevention efforts inside correctional facilities. These findings highlight the need for risk reduction programs for incarcerated men. PMID- 15539548 TI - Observational study in ten beauty salons: results informing development of the North Carolina BEAUTY and Health Project. AB - Researchers from the North Carolina BEAUTY and Health Project conducted an observational study in 10 North Carolina beauty salons to gain insight into naturally occurring conversations between cosmetologists and customers, and to assess features of the salon environment that might be used to inform the development of salon-based health promotion interventions. Results revealed that the social environment of a salon is a place where cosmetologists and customers talk openly about many subjects, including health. Information, advice, appraisal, humor, and empathy are typically shared in these health conversations. Several features of the physical environment of the salon may be mobilized to support health--access to healthy foods, snacks, and beverages; smoking restrictions; and availability of print or video materials, signs, or displays that include healthy messages. Implications for planning salon-based health promotion interventions--including the training of licensed cosmetologists to deliver health messages--are discussed in light of these findings. PMID- 15539549 TI - Assessing interorganizational networks as a dimension of community capacity: illustrations from a community intervention to prevent lead poisoning. AB - Network analysis is often cited as a method for assessing collaboration among organizations as an indicator of community capacity. The purpose of this study was to (1) document patterns of collaboration in organizational networks related to lead poisoning prevention in a Native American community and (2) examine measurement issues in using organizational network analysis to assess community capacity. Interviews were conducted with representatives from 22 tribes, government agencies, schools, and community-based organizations in northeastern Oklahoma. Intensity, density, and reliability were assessed for several stages of collaboration. Intensity and density were greater for similar types of organizations than for the network as a whole and decreased as stage of collaboration increased. Network data were more reliable when responses were dichotomized than when intensities were compared. Mean reliability scores across two respondents from the same organization ranged from 60% to 90%. Results from network studies may help communities learn how to strengthen organizational networks to enhance community capacity. PMID- 15539550 TI - Inhibition of the serotonin (5-hydroxytryptamine) transporter reduces bone accrual during growth. AB - Selective serotonin-reuptake inhibitors (SSRIs) antagonize the serotonin (5 hydroxytryptamine) transporter (5-HTT), and are frequently prescribed to children and adolescents to treat depression. However, recent findings of functional serotonergic pathways in bone cells and preliminary clinical evidence demonstrating detrimental effects of SSRIs on bone growth have raised questions regarding the effects of these drugs on the growing skeleton. The current work investigated the impact of 5-HTT inhibition on the skeleton in: 1) mice with a null mutation in the gene encoding for the 5-HTT; and 2) growing mice treated with a SSRI. In both models, 5-HTT inhibition had significant detrimental effects on bone mineral accrual. 5-HTT null mutant mice had a consistent skeletal phenotype of reduced mass, altered architecture, and inferior mechanical properties, whereas bone mineral accrual was impaired in growing mice treated with a SSRI. These phenotypes resulted from a reduction in bone formation without an increase in bone resorption and were not influenced by effects on skeletal mechanosensitivity or serum biochemistries. These findings indicate a role for the 5-HTT in the regulation of bone accrual in the growing skeleton and point to a need for further research into the prescription of SSRIs to children and adolescents. PMID- 15539551 TI - Bovine growth hormone transgenic mice are resistant to diet-induced obesity but develop hyperphagia, dyslipidemia, and diabetes on a high-fat diet. AB - It is known that bovine GH (bGH) transgenic mice have increased body mass, insulin resistance, and altered lipoprotein metabolism when fed a normal diet (ND). In this study, the effects of 8 wk of high-fat diet (HFD) were investigated in 6-month-old male bGH mice. Although littermate controls had unchanged energy intake, energy intake was higher in the bGH mice on a HFD than on a low-fat diet. Nevertheless, the bGH mice were resistant to diet-induced weight gain, and only in the bGH mice did the HFD result in increased energy expenditure. Glucose oxidation was higher in the bGH mice compared with littermate controls on both a HFD and ND. In addition, the bGH mice had 0.5 C higher body temperature throughout the day and increased hepatic uncoupling protein 2 expression; changes that were unaffected by the HFD. On a HFD, the effect of bGH overexpression on serum triglycerides and apolipoprotein B was opposite to that on a ND, resulting in higher serum concentrations of triglycerides and apolipoprotein B compared with littermate controls. Increased serum triglycerides were explained by decreased triglyceride clearance. The HFD led to diabetes only in the bGH mice. In conclusion, bGH transgenic mice were resistant to diet-induced obesity despite hyperphagia, possibly due to increased energy expenditure. On a HFD, bGH mice became dyslipidemic and diabetic and thereby more accurately reflect the metabolic situation in acromegalic patients. PMID- 15539553 TI - Leptin serves as an upstream activator of an obligatory signaling cascade in the embryo-implantation process. AB - Leptin is essential for mouse reproduction, but the exact roles it serves are yet to be determined. Treatment of cultured endometrial cells with leptin increases the level of beta3-integrin, IL-1, leukemia inhibitory factor, and their corresponding receptors. These leptin-induced effects are eliminated by inhibitors of leptin receptor (OB-R) signaling. Herein the impact of blocking leptin/OB-R signaling in the mouse endometrium was assessed. Intrauterine injection of either leptin peptide antagonists (LPA-1 or -2) or OB-R antibody on d 3 of pregnancy impaired mouse implantation in comparison to intrauterine injection of scrambled peptides (LPA-Sc) or species-matched IgGs. Significant reduction in the number of implantation sites and uterine horns with implanted embryos was found after intrauterine injection of LPA-1 (1 of 22) vs. LPA-1Sc (11 of 15) and LPA-2 (3 of 17) vs. LPA-2Sc (14 of 16). The impact of disruption of leptin signaling on the endometrial expression of several molecules in pregnant mice was assessed by Western blot, immunohistochemistry, and confocal microscopy. Disruption of leptin signaling resulted in a significant reduction of IL-1 receptor type I, leukemia inhibitory factor, vascular endothelial growth factor receptor 2, and beta3-integrin levels. The levels of colony stimulating factor-1 receptor and OB-R were unaltered after treatment with LPAs compared with controls. Expression of OB-R protein was pregnancy dependent and found only in glandular epithelium after implantation occurred. Our findings support previous observations that leptin signaling is critical to the implantation process and suggest that molecules downstream of leptin-activated receptor may serve obligatory roles in endometrial receptivity and successful implantation. PMID- 15539552 TI - Testosterone down-regulates ornithine aminotransferase gene and up-regulates arginase II and ornithine decarboxylase genes for polyamines synthesis in the murine kidney. AB - The enzymes ornithine aminotransferase (OAT) and ornithine decarboxylase (ODC) share L-ornithine as a common substrate and arginase II produces this amino acid. In the murine kidney, testosterone induced ODC gene expression and polyamine production, but it is unknown how OAT gene is expressed under androgen treatment. These experiments were designed to study the influence of testosterone on the renal expression of OAT gene. Pharmacological and physiological doses of testosterone were injected into female and castrated male mice. Total RNA and soluble proteins extracted from whole kidneys were analyzed by Northern and Western blots, respectively. The results clearly indicate that pharmacological doses of testosterone simultaneously down-regulated the level of OAT protein and up-regulated the expression of arginase II and ODC genes. Variations of the levels of OAT protein and arginase II mRNA and protein were strongly correlated with testosteronemia. Orchidectomy increased the renal level of OAT protein and decreased that of ODC and arginase II. These effects were reversed by injecting a physiological dose of testosterone into castrated male mice. In conclusion, OAT and ODC genes are inversely regulated by testosterone in the mouse kidney. Consequently, in kidneys of testosterone-treated mice, L-arginine-derived ornithine produced by arginase II might be preferentially used by ODC for putrescine production rather than by OAT. This metabolic fate of L-ornithine was facilitated by decreasing OAT gene expression. In contrast, in female and castrated male mice devoided of testosterone, OAT gene is highly expressed and L ornithine is converted into L-glutamate. PMID- 15539554 TI - Intravenous infusion of peptide YY(3-36) potently inhibits food intake in rats. AB - Peptide YY (3-36) [PYY (3-36)] is postulated to act as a hormonal signal from the gut to the brain to inhibit food intake and gastric emptying. A mixed-nutrient meal produces a prolonged 2-3 h increase in plasma levels of both PYY (3-36) and PYY (1-36). We determined the dose-dependent effects of 3-h iv infusions of PYY (3-36) and PYY (1-36) (0.5-50 pmol.kg(-1).min(-1)) at dark onset on food intake in non-food-deprived rats. PYY (3-36) dose-dependently inhibited food intake: the minimal effective dose was 5 pmol.kg(-1).min(-1); the estimated potency (mean effective dose) and efficacy (maximal percent inhibition) were 15 pmol.kg( 1).min(-1) (2.6 nmol/kg) and 47%, respectively. PYY (1-36) was an order of magnitude less potent than PYY (3-36). Similar total doses of PYY (3-36) (0.9-30 nmol/kg) infused during the 15-min period just before dark onset also dose dependently inhibited food intake, albeit with a lower potency and efficacy. Other experiments showed that PYY (3-36) inhibited food intake in sham-feeding rats and was more effective in reducing intake of a mixed-nutrient liquid diet than 15% aqueous sucrose. We conclude that: 1) iv infusions of PYY (3-36), which are more likely than ip injections to mimic postprandial increases in plasma PYY (3-36), potently inhibit food intake in a dose-dependent manner; 2) PYY (1-36) is an order of magnitude less potent than PYY (3-36); and 3) PYY (3-36) can inhibit food intake independently of its action to inhibit gastric emptying. It remains to be determined whether iv doses of PYY (3-36) that reproduce postprandial increases in plasma PYY (3-36) are sufficient to inhibit food intake. PMID- 15539555 TI - Bovine serum albumin-estrogen compounds differentially alter gonadotropin releasing hormone-1 neuronal activity. AB - Steroid hormones regulate a host of physiological processes and behaviors. These actions can occur by genomic mechanisms involving gene transcription or by nongenomic mechanisms proposed to involve receptors associated with the plasma membrane. BSA-conjugated steroid hormones have been extensively used to elucidate signal transduction pathways for these hormones. We have previously shown, using calcium imaging, that 17beta-estradiol (E2) significantly increases GnRH-1 neuronal activity. During the course of these experiments, it became apparent that three different BSA-estrogen compounds have been used in a variety of cell types: 17beta-estradiol 6-O-carboxymethyloxime-BSA (E2-6-BSA); 1,3,5(10) estratrien-3,16alpha,17beta-triol-6-one 6-O-carboxymethyloxime-BSA (E-6-BSA); and 1,3,5(10)-estratrien-3,17beta-diol 17-hemisuccinate-BSA (E2-17-BSA). The effects of these compounds on GnRH-1 neuronal activity were compared using calcium imaging. E-6-BSA and E2-17-BSA, but not E2-6-BSA, significantly increased all parameters of GnRH-1 neuronal activity. In addition, the effects of these two BSA compounds were reversed by the estrogen receptor antagonist ICI 182,780 but not by inhibition of gene transcription. The effects of E2-17-BSA, but not E-6-BSA were reversed by treatment with pertussis toxin, which blocks G protein-coupled receptors. These data indicate that these compounds cannot be used interchangeably and clearly have different binding properties and/or different effects on target tissues. PMID- 15539556 TI - Identity of an estrogen membrane receptor coupled to a G protein in human breast cancer cells. AB - Although nonclassical estrogen actions initiated at the cell surface have been described in many tissues, the identities of the membrane estrogen receptors (mERs) mediating these actions remain unclear. Here we show that GPR30, an orphan receptor unrelated to nuclear estrogen receptors, has all the binding and signaling characteristics of a mER. A high-affinity (dissociation constant 2.7 nm), limited capacity, displaceable, single binding site specific for estrogens was detected in plasma membranes of SKBR3 breast cancer cells that express GPR30 but lack nuclear estrogen receptors. Progesterone-induced increases and small interfering RNA-induced decreases in GPR30 expression in SKBR3 cells were accompanied by parallel changes in specific estradiol-17beta (E2) binding. Plasma membranes of human embryonic kidney 293 cells transfected with GPR30, but not those of untransfected cells, and human placental tissues that express GPR30 also displayed high-affinity, specific estrogen binding typical of mERs. E2 treatment of transfected cell membranes caused activation of a stimulatory G protein that is directly coupled to the receptor, indicating GPR30 is a G protein-coupled receptor (GPCR), and also increased adenylyl cyclase activity. The finding that the antiestrogens tamoxifen and ICI 182,780, and an environmental estrogen, ortho,para-dichlorodiphenyldichloroethylene (o,p'-DDE), have high binding affinities to the receptor and mimic the actions of E2 has important implications for both the development and treatment of estrogen-dependent breast cancer. GPR30 is structurally unrelated to the recently discovered family of GPCR-like membrane progestin receptors. The identification of a second distinct class of GPCR-like steroid membrane receptors suggests a widespread role for GPCRs in nonclassical steroid hormone actions. PMID- 15539557 TI - Angiotensin II stimulates protein synthesis and inhibits proliferation in primary cultures of rat adrenal glomerulosa cells. AB - Angiotensin II (Ang II) is one of the most important stimuli of rat adrenal glomerulosa cells. The aim of the present study was to investigate whether Ang II can stimulate cell proliferation and/or hypertrophy and investigate pathways and intracellular targets. A 3-d treatment with Ang II (5-100 nm), through the Ang II type 1 receptor subtype, abolished cell proliferation observed in control cells but increased protein synthesis. Preincubation with PD98059 (a MAPK kinase inhibitor) abolished basal proliferation and had no effect on basal protein synthesis but did reverse the effect of Ang II on protein synthesis. The p38 MAPK inhibitor SB203580 reversed the inhibitory effect on cell proliferation and abolished the increase in protein synthesis, whereas the c-Jun N-terminal kinase inhibitor SP600125 had no effect. Time-course studies revealed that Ang II stimulated phosphorylation of both p42/p44mapk and p38 MAPK but did not activate c-Jun N-terminal kinase. Ang II had no effect on the level of cyclin E expression but increased the expression of the cyclin-dependent kinase, p27Kip1, an effect abolished in cells preincubated with SB203580 and PD98059. In conclusion, in cultured rat glomerulosa cells, a 3-d treatment with Ang II increases protein synthesis, with a concomitant decrease in proliferation. These effects are mediated by both the p42/p44mapk and p38 MAPK pathways, which increase expression of the steroidogenic enzymes, steroidogenic acute regulatory protein and 3beta hydroxysteroid dehydrogenase and p27Kip1, a protein known to block the cell cycle in G1 phase. Together these results support the key role of Ang II as a stimulus of steroid synthesis rather than a proliferating factor. PMID- 15539559 TI - Follicle-stimulating hormone regulates oocyte growth by modulation of expression of oocyte and granulosa cell factors. AB - Oocyte-granulosa cell communication is essential for oocyte development. The aims of this study were: 1) to determine the effect of FSH on expression of Kit ligand (KL), growth/differentiation factor-9, bone morphogenetic protein (BMP)-15, and Kit during growth of oocyte-granulosa cell complexes (OGCs) in vitro; 2) to investigate the role of BMP-15 in regulation of KL expression; and 3) to correlate mRNA expression with oocyte growth. OGCs from 12-d-old mice were cultured for up to 7 d in the presence of FSH [0.05 ng/ml (low), 5 ng/ml (high)] or BMP-15 (10 or 100 ng/ml). Transcripts were quantified using real-time RT-PCR, and oocyte and OGC diameters were measured. FSH regulated KL expression in a biphasic manner, with low FSH decreasing the KL-1/KL-2 ratio, and high FSH increasing the KL-1/KL-2 ratio, compared with controls (P < 0.05). The decrease in KL-1/KL-2 ratio with low FSH was due to increased KL-2 mRNA expression. Both FSH concentrations increased OGC diameter (P < 0.05), but only low FSH promoted oocyte growth (P < 0.05). High FSH also decreased BMP-15 expression (P < 0.05). FSH-stimulated oocyte growth was inhibited by Gleevec, an inhibitor of Kit activity. BMP-15 increased both KL-1 and KL-2 mRNA levels in a dose-dependent manner (P < 0.05) but did not alter the KL-1/KL-2 ratio or promote oocyte growth. When the KL-1/KL-2 ratio was increased by exogenous KL-1, FSH-stimulated oocyte growth was suppressed (P < 0.05), suggesting that lowered KL-1/KL-2 ratio is important for oocyte growth. In summary, the correct concentration of FSH is crucial for appropriate modulation of KL and BMP-15 to promote oocyte growth. PMID- 15539558 TI - Miniglucagon (MG)-generating endopeptidase, which processes glucagon into MG, is composed of N-arginine dibasic convertase and aminopeptidase B. AB - Miniglucagon (MG), the C-terminal glucagon fragment, processed from glucagon by the MG-generating endopeptidase (MGE) at the Arg17-Arg18 dibasic site, displays biological effects opposite to that of the mother-hormone. This secondary processing occurs in the glucagon- and MG-producing alpha-cells of the islets of Langerhans and from circulating glucagon. We first characterized the enzymatic activities of MGE in culture media from glucagon and MG-secreting alphaTC1.6 cells as made of a metalloendoprotease and an aminopeptidase. We observed that glucagon is a substrate for N-arginine dibasic convertase (NRDc), a metalloendoprotease, and that aminopeptidase B cleaves in vitro the intermediate cleavage products sequentially, releasing mature MG. Furthermore, immunodepletion of either enzyme resulted in the disappearance of the majority of MGE activity from the culture medium. We found RNAs and proteins corresponding to both enzymes in different cell lines containing a MGE activity (mouse alphaTC1.6 cells, rat hepatic FaO, and rat pituitary GH4C1). Using confocal microscopy, we observed a granular immunostaining of both enzymes in the alphaTC1.6 and native rat alpha cells from islets of Langerhans. By immunogold electron microscopy, both enzymes were found in the mature secretory granules of alpha-cells, close to their substrate (glucagon) and their product (MG). Finally, we found NRDc only in the fractions from perfused pancreas that contain glucagon and MG after stimulation by hypoglycemia. We conclude that MGE is composed of NRDc and aminopeptidase B acting sequentially, providing a molecular basis for this uncommon regulatory process, which should be now addressed in both physiological and pathophysiological situations. PMID- 15539560 TI - Skeletal overexpression of gremlin impairs bone formation and causes osteopenia. AB - Skeletal cells synthesize bone morphogenetic proteins (BMPs) and BMP antagonists. Gremlin, a BMP antagonist, is expressed in osteoblasts and opposes BMP effects on osteoblastic differentiation and function in vitro. However, its effects in vivo are not known. To investigate the actions of gremlin on bone remodeling in vivo, we generated transgenic mice overexpressing gremlin under the control of the osteocalcin promoter. Gremlin transgenics exhibited bone fractures and reduced bone mineral density by 20-30%, compared with controls. Static and dynamic histomorphometry of femurs revealed that gremlin overexpression caused reduced trabecular bone volume and the appearance of woven bone. Polarized light microscopy revealed disorganized collagen bundles at the endosteal cortical surface. Gremlin transgenic mice displayed a 70% decrease in the number of osteoblasts/trabecular area and reduced mineral apposition and bone formation rates. In vivo bromodeoxyuridine labeling and marrow stromal cell cultures demonstrated an inhibitory effect of gremlin on osteoblastic cell replication, but no change on apoptosis was detected. Marrow stromal cells from gremlin transgenics displayed a reduced response to BMP on phosphorylated mothers against decapentaplegic 1/5/8 phosphorylation and reduced free cytosolic beta-catenin levels. In conclusion, transgenic mice overexpressing gremlin in the bone microenvironment have decreased osteoblast number and function leading to osteopenia and spontaneous fractures. PMID- 15539561 TI - Abnormal Leydig Cell aggregation in the fetal testis of rats exposed to di (n butyl) phthalate and its possible role in testicular dysgenesis. AB - Fetal exposure of male rats to di (n-butyl) phthalate (DBP) induces testicular changes remarkably similar to testicular dysgenesis syndrome in humans; these include induction of focal areas of dysgenetic tubules in otherwise normal testes. In searching for the fetal origins of the latter, we used image analysis to show that exposure to 500 mg/kg DBP [embryonic day (E)13.5-20.5)] caused abnormal aggregation of Leydig cells centrally in the fetal testis. This aggregation was not due to increase in Leydig cell number, and Leydig cell size was significantly reduced in DBP-exposed animals, as were testosterone levels and immunoexpression of P450 side-chain cleavage enzyme. The Leydig cell aggregates did not exhibit evidence of focal proliferation at E17.5-19.5. Using confocal microscopy and Leydig (3beta-hydroxysteroid dehydrogenase) and Sertoli (anti Mullerian hormone) cell-specific markers, we show that fetal Leydig cell aggregates in DBP-exposed animals trap isolated Sertoli cells within them at E21.5. These areas of intermingled cells are still apparent on postnatal d 4, after cessation of DBP treatment, when they may form misshapen seminiferous cords that trap (intratubular) Leydig cells within them. These centrally located dysgenetic tubules contain germ cells in early puberty, but by adulthood they are Sertoli cell only, implying that presence of intratubular Leydig cells interferes with spermatogenesis. It is concluded that DBP-induced fetal Leydig cell aggregation may be a key event in formation of focal dysgenetic areas in the testis, and identification of the mechanisms underlying these events may give new insights into the fetal origins of testicular dysgenesis syndrome disorders in the human. PMID- 15539562 TI - Duplicate zebrafish pth genes are expressed along the lateral line and in the central nervous system during embryogenesis. AB - PTH plays a critical role in calcium metabolism in tetrapods. The primary site of PTH expression is the parathyroid glands, although it is also detected in the thymus and hypothalamus. Fish lack anatomically distinct parathyroid glands, and the first animals to evolve parathyroid glands were the amphibians. However, fish do have PTH family ligands and receptors, which are functionally similar to their mammalian counterparts. We report the expression patterns of duplicate zebrafish pth genes during embryogenesis. Both zebrafish pth1 and pth2 transcripts are expressed along the lateral line before the migration of the lateral line primordium and later in development Pth protein is detected in lateral line neuromasts by immunohistochemistry. pth1 transcripts are also detected in the central nervous system in the ventral neural tube. These temporally and anatomically restricted expression patterns imply a novel role for PTH family hormones during embryonic development of the zebrafish and allow for the genetic dissection of PTH function in this model organism. PMID- 15539563 TI - Observation of the spin Hall effect in semiconductors. AB - Electrically induced electron-spin polarization near the edges of a semiconductor channel was detected and imaged with the use of Kerr rotation microscopy. The polarization is out-of-plane and has opposite sign for the two edges, consistent with the predictions of the spin Hall effect. Measurements of unstrained gallium arsenide and strained indium gallium arsenide samples reveal that strain modifies spin accumulation at zero magnetic field. A weak dependence on crystal orientation for the strained samples suggests that the mechanism is the extrinsic spin Hall effect. PMID- 15539564 TI - Human prion protein with valine 129 prevents expression of variant CJD phenotype. AB - Variant Creutzfeldt-Jakob disease (vCJD) is a unique and highly distinctive clinicopathological and molecular phenotype of human prion disease associated with infection with bovine spongiform encephalopathy (BSE)-like prions. Here, we found that generation of this phenotype in transgenic mice required expression of human prion protein (PrP) with methionine 129. Expression of human PrP with valine 129 resulted in a distinct phenotype and, remarkably, persistence of a barrier to transmission of BSE-derived prions on subpassage. Polymorphic residue 129 of human PrP dictated propagation of distinct prion strains after BSE prion infection. Thus, primary and secondary human infection with BSE-derived prions may result in sporadic CJD-like or novel phenotypes in addition to vCJD, depending on the genotype of the prion source and the recipient. PMID- 15539566 TI - Global identification of human transcribed sequences with genome tiling arrays. AB - Elucidating the transcribed regions of the genome constitutes a fundamental aspect of human biology, yet this remains an outstanding problem. To comprehensively identify coding sequences, we constructed a series of high density oligonucleotide tiling arrays representing sense and antisense strands of the entire nonrepetitive sequence of the human genome. Transcribed sequences were located across the genome via hybridization to complementary DNA samples, reverse transcribed from polyadenylated RNA obtained from human liver tissue. In addition to identifying many known and predicted genes, we found 10,595 transcribed sequences not detected by other methods. A large fraction of these are located in intergenic regions distal from previously annotated genes and exhibit significant homology to other mammalian proteins. PMID- 15539565 TI - Lysosomal glycosphingolipid recognition by NKT cells. AB - NKT cells represent a distinct lineage of T cells that coexpress a conserved alphabeta T cell receptor (TCR) and natural killer (NK) receptors. Although the TCR of NKT cells is characteristically autoreactive to CD1d, a lipid-presenting molecule, endogenous ligands for these cells have not been identified. We show that a lysosomal glycosphingolipid of previously unknown function, isoglobotrihexosylceramide (iGb3), is recognized both by mouse and human NKT cells. Impaired generation of lysosomal iGb3 in mice lacking beta-hexosaminidase b results in severe NKT cell deficiency, suggesting that this lipid also mediates development of NKT cells in the mouse. We suggest that expression of iGb3 in peripheral tissues may be involved in controlling NKT cell responses to infections and malignancy and in autoimmunity. PMID- 15539567 TI - What's on the label? PMID- 15539568 TI - U.S. science policy. Bush victory leaves scars--and concerns about funding. PMID- 15539570 TI - U.S. higher education. Decline in new foreign grad students slows. PMID- 15539569 TI - U.S. science policy. California's Proposition 71 launches stem cell gold rush. PMID- 15539571 TI - Ocean science. NOAA to retool research programs. PMID- 15539572 TI - Scientific publishing. Mixed week for open access in the U.K. PMID- 15539574 TI - Condensed matter physics. Spin current sighting ends 35-year hunt. PMID- 15539573 TI - Paleoanthropology. Skeptics question whether Flores hominid is a new species. PMID- 15539575 TI - Medicine. Sperm-targeting vaccine blocks male fertility in monkeys. PMID- 15539576 TI - Drug testing. Regulators talk up plans for drug biomarkers ... PMID- 15539577 TI - Cancer research. ...and NCI hears a pitch for biomarker studies. PMID- 15539578 TI - Evolution of behavior. Seeking the key to music. PMID- 15539579 TI - Vaccine policy. Immunizing kids against flu may prevent deaths among the elderly. PMID- 15539580 TI - Molecular biology. RNAi shows cracks in its armor. PMID- 15539581 TI - Society for Neuroscience meeting. Brain cells may pay the price for a bad night's sleep. PMID- 15539582 TI - Society for Neuroscience meeting. Anesthesia's addiction problem. PMID- 15539584 TI - The future of NASA. PMID- 15539583 TI - Society for Neuroscience meeting. Listen, baby. PMID- 15539585 TI - Research Corporation and John Schaefer. PMID- 15539587 TI - Oxide surface films on metal crystals. PMID- 15539586 TI - The file-drawer problem, revisited. PMID- 15539588 TI - Protein design: quo vadis? PMID- 15539589 TI - Ecology. How extinction patterns affect ecosystems. PMID- 15539590 TI - Physics. Superfluid helium-3 has a metallic partner. PMID- 15539591 TI - Ocean science. Deep ocean overturning--then and now. PMID- 15539592 TI - Evolution. Genomic databases and the tree of life. PMID- 15539593 TI - Biodiversity conservation and the eradication of poverty. AB - It is widely accepted that biodiversity loss and poverty are linked problems and that conservation and poverty reduction should be tackled together. However, success with integrated strategies is elusive. There is sharp debate about the social impacts of conservation programs and the success of community-based approaches to conservation. Clear conceptual frameworks are needed if policies in these two areas are to be combined. We review the links between poverty alleviation and biodiversity conservation and present a conceptual typology of these relationships. PMID- 15539594 TI - Pleistocene brown bears in the mid-continent of North America. AB - Current biogeographic models hypothesize that brown bears migrated from Asia to the New World ~100 to 50 thousand years ago but did not reach areas south of Beringia until ~13 to 12 thousand years ago, after the opening of a mid continental ice-free corridor. We report a 26-thousand-year-old brown bear fossil from central Alberta, well south of Beringia. Mitochondrial DNA recovered from the specimen shows that it belongs to the same clade of bears inhabiting southern Canada and the northern United States today and that modern brown bears in this region are probably descended from populations that persisted south of the southern glacial margin during the Last Glacial Maximum. PMID- 15539595 TI - Odd-parity superconductivity in Sr2RuO4. AB - Phase-sensitive measurements were made on Sr2RuO4 to establish unambiguously the odd-parity pairing in this material. The critical current of Au(0.5)In(0.5) Sr2RuO4 superconducting quantum interference devices prepared on Sr2RuO4 single crystals was found to be a maximum for devices with junctions on the same side of the crystal and a minimum for devices with junctions on opposite sides, in the limit of zero magnetic flux; these findings indicate that the phase of the superconducting order parameter in Sr2RuO4 changes by pi under inversion. This result verifies the odd-parity pairing symmetry and the formation of spin-triplet Cooper pairs in Sr2RuO4. PMID- 15539596 TI - Disorder-sensitive phase formation linked to metamagnetic quantum criticality. AB - Condensed systems of strongly interacting electrons are ideal for the study of quantum complexity. It has become possible to promote the formation of new quantum phases by explicitly tuning systems toward special low-temperature quantum critical points. So far, the clearest examples have been appearances of superconductivity near pressure-tuned antiferromagnetic quantum critical points. We present experimental evidence for the formation of a nonsuperconducting phase in the vicinity of a magnetic field-tuned quantum critical point in ultrapure crystals of the ruthenate metal Sr3Ru2O7, and we discuss the possibility that the observed phase is due to a spin-dependent symmetry-breaking Fermi surface distortion. PMID- 15539597 TI - Sea urchin spine calcite forms via a transient amorphous calcium carbonate phase. AB - The skeletons of adult echinoderms comprise large single crystals of calcite with smooth convoluted fenestrated morphologies, raising many questions about how they form. By using water etching, infrared spectroscopy, electron diffraction, and environmental scanning electron microscopy, we show that sea urchin spine regeneration proceeds via the initial deposition of amorphous calcium carbonate. Because most echinoderms produce the same type of skeletal material, they probably all use this same mechanism. Deposition of transient amorphous phases as a strategy for producing single crystals with complex morphology may have interesting implications for the development of sophisticated materials. PMID- 15539598 TI - Ventilation of the glacial deep Pacific Ocean. AB - Measurements of the age difference between coexisting benthic and planktic foraminifera from western equatorial Pacific deep-sea cores suggest that during peak glacial time the radiocarbon age of water at 2-kilometers depth was no greater than that of today. These results make unlikely suggestions that a slowdown in deep-ocean ventilation was responsible for a sizable fraction of the increase of the ratio of carbon-14 (14C) to carbon in the atmosphere and surface ocean during glacial time. Comparison of 14C ages for coexisting wood and planktic foraminifera from the same site suggests that the atmosphere to surface ocean 14C to C ratio difference was not substantially different from today's. PMID- 15539599 TI - Prospects for building the tree of life from large sequence databases. AB - We assess the phylogenetic potential of approximately 300,000 protein sequences sampled from Swiss-Prot and GenBank. Although only a small subset of these data was potentially phylogenetically informative, this subset retained a substantial fraction of the original taxonomic diversity. Sampling biases in the databases necessitate building phylogenetic data sets that have large numbers of missing entries. However, an analysis of two "supermatrices" suggests that even data sets with as much as 92% missing data can provide insights into broad sections of the tree of life. PMID- 15539600 TI - Realistic species losses disproportionately reduce grassland resistance to biological invaders. AB - Consequences of progressive biodiversity declines depend on the functional roles of individual species and the order in which species are lost. Most studies of the biodiversity-ecosystem functioning relation tackle only the first of these factors. We used observed variation in grassland diversity to design an experimental test of how realistic species losses affect invasion resistance. Because entire plant functional groups disappeared faster than expected by chance, resistance declined dramatically with progressive species losses. Realistic biodiversity losses, even of rare species, can thus affect ecosystem processes far more than indicated by randomized-loss experiments. PMID- 15539601 TI - Extinction and ecosystem function in the marine benthos. AB - Rapid changes in biodiversity are occurring globally, yet the ecological impacts of diversity loss are poorly understood. Here we use data from marine invertebrate communities to parameterize models that predict how extinctions will affect sediment bioturbation, a process vital to the persistence of aquatic communities. We show that species extinction is generally expected to reduce bioturbation, but the magnitude of reduction depends on how the functional traits of individual species covary with their risk of extinction. As a result, the particular cause of extinction and the order in which species are lost ultimately govern the ecosystem-level consequences of biodiversity loss. PMID- 15539602 TI - Bushmeat hunting, wildlife declines, and fish supply in West Africa. AB - The multibillion-dollar trade in bushmeat is among the most immediate threats to the persistence of tropical vertebrates, but our understanding of its underlying drivers and effects on human welfare is limited by a lack of empirical data. We used 30 years of data from Ghana to link mammal declines to the bushmeat trade and to spatial and temporal changes in the availability of fish. We show that years of poor fish supply coincided with increased hunting in nature reserves and sharp declines in biomass of 41 wildlife species. Local market data provide evidence of a direct link between fish supply and subsequent bushmeat demand in villages and show bushmeat's role as a dietary staple in the region. Our results emphasize the urgent need to develop cheap protein alternatives to bushmeat and to improve fisheries management by foreign and domestic fleets to avert extinctions of tropical wildlife. PMID- 15539603 TI - The genetic basis of singlet oxygen-induced stress responses of Arabidopsis thaliana. AB - Plants under oxidative stress suffer from damages that have been interpreted as unavoidable consequences of injuries inflicted upon plants by toxic levels of reactive oxygen species (ROS). However, this paradigm needs to be modified. Inactivation of a single gene, EXECUTER1, is sufficient to abrogate stress responses of Arabidopsis thaliana caused by the release of singlet oxygen: External conditions under which these stress responses are observed and the amounts of ROS that accumulate in plants exposed to these environmental conditions do not directly cause damages. Instead, seedling lethality and growth inhibition of mature plants result from genetic programs that are activated after the release of singlet oxygen has been perceived by the plant. PMID- 15539604 TI - Microbial factor-mediated development in a host-bacterial mutualism. AB - Tracheal cytotoxin (TCT), a fragment of the bacterial surface molecule peptidoglycan (PGN), is the factor responsible for the extensive tissue damage characteristic of whooping cough and gonorrhea infections. Here, we report that Vibrio fischeri also releases TCT, which acts in synergy with lipopolysaccharide (LPS) to trigger tissue development in its mutualistic symbiosis with the squid Euprymna scolopes. As components of PGN and LPS have commonly been linked with pathogenesis in animals, these findings demonstrate that host interpretation of these bacterial signal molecules is context dependent. Therefore, such differences in interpretation can lead to either inflammation and disease or to the establishment of a mutually beneficial animal-microbe association. PMID- 15539605 TI - Reversible immunocontraception in male monkeys immunized with eppin. AB - Various forms of birth control have been developed for women; however, there are currently few options for men. The development of male contraceptives that are effective, safe, and reversible is desired for family planning throughout the world. We now report contraception of male nonhuman primates (Macaca radiata) immunized with Eppin, a testis/epididymis-specific protein. Seven out of nine males (78%) developed high titers to Eppin, and all of these high-titer monkeys were infertile. Five out of seven (71%) high-anti-Eppin titer males recovered fertility when immunization was stopped. This study demonstrates that effective and reversible male immunocontraception is an attainable goal. This method of immunocontraception may be extended to humans. PMID- 15539606 TI - Multidimensional drug profiling by automated microscopy. AB - We present a method for high-throughput cytological profiling by microscopy. Our system provides quantitative multidimensional measures of individual cell states over wide ranges of perturbations. We profile dose-dependent phenotypic effects of drugs in human cell culture with a titration-invariant similarity score (TISS). This method successfully categorized blinded drugs and suggested targets for drugs of uncertain mechanism. Multivariate single-cell analysis is a starting point for identifying relationships among drug effects at a systems level and a step toward phenotypic profiling at the single-cell level. Our methods will be useful for discovering the mechanism and predicting the toxicity of new drugs. PMID- 15539607 TI - The effects of selective and nonselective cyclooxygenase inhibitors on endothelin 1-induced fever in rats. AB - It was previously shown that sustained fever can be induced in rats by central injection of endothelin-1 (ET-1). This peptide appears to participate in the mechanism(s) of LPS-induced fever, which is reduced by pretreatments with ET(B) receptor antagonists. In this study, we compared the effects of a nonselective cyclooxygenase (COX) inhibitor, indomethacin, with those of two selective COX-2 inhibitors, celecoxib and lumiracoxib, on ET-1-induced fever in rats. Fever induced in conscious animals by ET-1 (1 pmol icv) or LPS (5 mug/kg iv) was prevented by pretreatments with celecoxib (5 and 10 mg/kg) or lumiracoxib (5 mg/kg) given by oral gavage 1 h before stimuli. Lower doses of celecoxib had partial (2.5 mg/kg) or no effect (1 mg/kg). Indomethacin (2 mg/kg ip) partially inhibited fever induced by LPS but had no effect on ET-1-induced fever. The levels of PGE(2) and PGF(2alpha) in the cerebrospinal fluid (CSF) of pentobarbital sodium-anesthetized rats were significantly increased 3 h after the injection of LPS or ET-1. The latter increase was abolished by celecoxib at all tested doses and by indomethacin. In conclusion, selective COX-2 inhibitors were able to prevent ET-1-induced fever, indicating a role for COX-2 in this phenomenon. However, the fact that reduced CSF PG levels obtained with indomethacin and a low dose of celecoxib are not accompanied by changes in fever induced by ET-1, along with the lack of inhibitory effects of indomethacin on ET 1 fever, suggests that the latter might also involve COX-2-independent mechanisms. PMID- 15539608 TI - Uterine motor alterations and estrous cycle disturbances associated with colonic inflammation in the rat. AB - The impact of colitis on uterine contractility and estrous cycle was investigated after intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid (TNBS) in rats. Colitis severity was assessed by macroscopic damage scoring (MDS) 4 days after TNBS, and myeloperoxidase (MPO) activity was measured in both colon and uterus of control and colitic rats. Estrous cycle stages were determined by vaginal smears and histology, and uterine contractility was assessed in vitro on longitudinal and circular strips. In control rats, uterine MPO activity varied markedly during the cycle and peaked around estrus. In rats with moderate colitis [MDS < 5, 3.1 +/- 0.2 (mean +/- SE)], uterine MPO decreased by 61% compared with estrus control, without disruption of the cycle. Frequency of spontaneous contractions was reduced by 32% in circular muscle. Contractile responses to KCl and carbachol were not affected, whereas maximal response to oxytocin decreased by 47% in the longitudinal muscle. In rats with severe colitis (MDS > 5, 6.0 +/- 0.2), uterine MPO was reduced by 96% and estrous cycle was disrupted. Spontaneous contractility was impaired in circular strips, and a 39% decrease in the contraction frequency occurred in the longitudinal strips. Circular strips did not contract to KCl or carbachol; however, longitudinal strips had maximal responses to KCl, carbachol, and oxytocin reduced by 36%, 27%, and 46%, respectively. Estrogen replacement protected the uterine responses to carbachol in colitic rats, whereas oxytocin responses remained depressed. These data indicate that colonic inflammation can influence both spontaneous and evoked uterine contractility, in relation to estrous cycle disturbances, impaired estradiol production, and functional alterations of myometrial cells. PMID- 15539609 TI - Electric field strength of membrane lipids from vertebrate species: membrane lipid composition and Na+-K+-ATPase molecular activity. AB - Intramembrane electric field strength is a very likely determinant of the activity of ion-transporting membrane proteins in living cells. In the absence of any transmembrane electrical potential or surface potential, its magnitude is determined by the dipole potential of the membrane's lipid components and their associated water of hydration. Here we have used a fluorometric method to quantify the dipole potential of vesicles formed from lipids extracted from kidney and brain of 11 different animal species from four different vertebrate classes. The dipole potential was compared with the fatty acid composition and with the Na(+)-K(+)-ATPase molecular activity of each preparation. The magnitude of the dipole potential was found to be relatively constant across all animal species, i.e., 236-334 mV for vesicles prepared from the total membrane lipids and 223-256 mV for phospholipids alone. The significantly lower value for phospholipids alone is potentially related to the removal of cholesterol and/or other common soluble lipid molecules from the membrane. Surprisingly, no significant dependence of the dipole potential on fatty acid composition was found. This may, however, be due to concomitant compensatory variations in lipid head group composition. The molecular activity of the Na(+)-K(+)-ATPase was found to increase with increasing dipole potential. The fact that the dipole potential is maintained at a relatively constant value over a wide range of animal species suggests that it may play a fundamental role in ensuring correct ion pump conformation and function within the membrane. PMID- 15539610 TI - Hormonal regulation of glucose and system A amino acid transport in first trimester placental villous fragments. AB - Alterations in placental nutrient transfer have been implicated in fetal growth abnormalities. In pregnancies complicated by diabetes and accelerated fetal growth, upregulations of glucose transporter 1 (GLUT1) and amino acid transporter system A have been shown in the syncytiotrophoblast of term placenta. In contrast, intrauterine growth restriction is associated with a downregulation of placental system A transporters. However, underlying mechanisms of transporter regulation are poorly understood, particularly in early pregnancy. In this study, hormonal regulation of placental glucose and system A transporters was investigated. The uptake of 3-O-[methyl-(14)C]-d-glucose was studied in villous fragments isolated from first trimester (6-13 wk of gestation) and term human placenta. Villous fragments were incubated in buffer containing insulin, leptin, cortisol, growth hormone (GH), prolactin, IGF-I, or under hypo/hyperglycemic conditions for 1 h. Subsequently, 3-O-[methyl-(14)C]-D-glucose uptake was measured with and without phloretin for 70 s in first trimester tissue and 20 s in term tissue. Methylaminoisobutyric uptake was measured with and without Na+ for 20 min. Glucose uptake was unaltered by hormones or hypo/hyperglycemia. GH decreased system A activity by 31% in first trimester (P < 0.05). The uptake of glucose was 50% higher in term compared with first trimester fragments and increased markedly between 6 and 13 wk of gestation (P < 0.05). We conclude that placental glucose transporter activity is not regulated by short exposures to the hormones or glucose concentrations tested. In contrast to term placental villous fragments, system A activity was not regulated by insulin or leptin in first trimester but was downregulated by GH. PMID- 15539611 TI - Effects of acute hyperinsulinemia on insulin signal transduction and glucose transporters in ovine fetal skeletal muscle. AB - To test the effects of acute fetal hyperinsulinemia on the pattern and time course of insulin signaling in ovine fetal skeletal muscle, we measured selected signal transduction proteins in the mitogenic, protein synthetic, and metabolic pathways in the skeletal muscle of normally growing fetal sheep in utero. In experiment 1, 4-h hyperinsulinemic-euglycemic clamps were conducted in anesthetized twin fetuses to produce selective fetal hyperinsulinemia-euglycemia in one twin and euinsulinemia-euglycemia in the other. Serial skeletal muscle biopsies were taken from each fetus during the clamp and assayed by Western blot for selected insulin signal transduction proteins. Tyrosine phosphorylation of the insulin receptor, insulin receptor substrate-1, and the p85 subunit of phosphatidylinositol 3-kinase doubled at 30 min and gradually returned to control values by 240 min. Phosphorylation of extracellular signal-regulated kinase 1,2 was increased fivefold through 120 min of insulin infusion and decreased to control concentration by 240 min. Protein kinase B phosphorylation doubled at 30 min and remained elevated throughout the study. Phosphorylation of p70 S6K increased fourfold at 30, 60, and 120 min. In the second experiment, a separate group of nonanesthetized singleton fetuses was clamped to intermediate and high hyperinsulinemic-euglycemic conditions for 1 h. GLUT4 increased fourfold in the plasma membrane at 1 h, and hindlimb glucose uptake increased significantly at the higher insulin concentration. These data demonstrate that an acute increase in fetal plasma insulin concentration stimulates a unique pattern of insulin signal transduction proteins in intact skeletal muscle, thereby increasing pathways for mRNA translation, glucose transport, and cell growth. PMID- 15539612 TI - Tyrosine kinase inhibition affects skate anion exchanger isoform I alterations after volume expansion. AB - Upon exposure to hypotonic medium, skate red blood cells swell and then reduce their volume by releasing organic osmolytes and associated water. The regulatory volume decrease is inhibited by stilbenes and anion exchange inhibitors, suggesting involvement of the red blood cell anion exchanger skAE1. To determine the role of tyrosine phosphorylation, red blood cells were volume expanded with and without prior treatment with the tyrosine kinase inhibitor piceatannol. At the concentration used, 130 microM, piceatannol nearly completely inhibits p72(syk), a tyrosine kinase previously shown to phosphorylate skAE1 (M. W. Musch, E. H. Hubert, and L. Goldstein. J Biol Chem 274: 7923-7928, 1999). Hyposmotic induced volume expansion stimulated association of p72(syk) with a light membrane fraction of skate red blood cells. Piceatannol did not inhibit this association but decreased hyposmotically stimulated increased skAE1 tyrosine phophorylation. Movement of skAE1 from an intracellular to a surface detergent-resistant membrane domain and tetramer formation were not inhibited by piceatannol treatment. Two effects of hyposmotic-induced volume expansion, decreased band 4.1 binding and increased ankyrin, were both inhibited by piceatannol. These results suggest that at least one event requiring p72(syk) activation is pivotal for hyposmotic induced increased transport; however, steps that do not require tyrosine phosphorylation may also play a role. PMID- 15539613 TI - Modeling a hydropic recipient twin in twin-twin transfusion syndrome. AB - We developed a mathematical model of twin-twin transfusion syndrome (TTTS) that includes a hydropic recipient twin, adding interstitial and intracellular fluid compartments, fetal congestive cardiac failure, and the dynamics of renin angiotensin system (RAS) mediators to our previous TTTS model. Ten differential equations for each twin, coupled by the net fetofetal transfusion of blood and blood components, i.e., colloids, osmoles, and RAS mediators, describe the development of fetal arterial and venous blood volumes, blood osmolality and colloid osmotic pressure (COP), interstitial fluid volume and COP, intracellular fluid volume, amniotic fluid volume and osmolality, and RAS mediator concentration. We included varying placental anastomoses, placental sharing, and amnionicity. The 20 differential equations were solved numerically from 0 to 40 wk with a 0.6-s time step. Consistent with clinical experience, model predictions are as follows. Unidirectional arteriovenous anastomoses and arteriovenous anastomoses inadequately compensated by oppositely directed anastomoses cause severe TTTS that includes a hydropic recipient. Adequately compensated arteriovenous anastomoses simulated TTTS without hydrops. The probability that oppositely directed anastomoses prevent onset of a hydropic recipient after TTTS onset, i.e., the largest interval between onset of TTTS and onset of hydrops in the recipient, was best for a venovenous anastomosis, closely followed by an arterioarterial and finally an oppositely directed arteriovenous anastomosis. Hydropic recipients have decreased amniotic fluid volume. Unequal placental sharing and amnionicity modify hydrops onset. In conclusion, our model simulates a sequence of events that results in a hydropic recipient twin in severe TTTS. The model may allow an assessment of the efficacy of current therapeutic interventions for TTTS cases that include a hydropic recipient twin. PMID- 15539614 TI - Electrophysiological-anatomic correlates of ATP-triggered vagal reflex in the dog. V. Role of purinergic receptors. AB - The mechanism of extracellular ATP-triggered vagal depressor reflex was further studied in a closed-chest canine model. Adenosine and ATP were administered individually in equimolar doses (0.01-1.0 mumol/kg) into the right coronary artery (RCA) and left circumflex coronary artery (LCA). When administered into the RCA, adenosine and ATP exerted an identical and relatively small negative chronotropic effect on sinus node automaticity; the time to peak negative chronotropic effect was >/=7 s. When administered into the LCA, adenosine had no effect on sinus node automaticity, whereas ATP markedly suppressed sinus node automaticity. This effect of ATP 1) reached its peak in <2 s after its administration, 2) was short lasting, and 3) was completely abolished by either intravenous administration of the muscarinic cholinergic blocker atropine (0.2 mg/kg) or intra-LCA administration of 2',3'-O-(2,4,6-trinitrophenyl)-ATP (TNP ATP), a potent P2X(2/3) purinergic receptor (P2X(2/3)R) antagonist, but not by diinosine pentaphosphate (Ip(5)I), a potent inhibitor of P2X(1)R and P2X(3)R. Repetitive administrations of ATP were not associated with reduced effects, indicative of receptor desensitization, thereby excluding the involvement of the rapidly desensitized P2X(1)R in the action of ATP. It was concluded that ATP triggers a cardio-cardiac vagal depressor reflex by activating P2X(2/3)R located on vagal sensory nerve terminals localized in the left ventricle. Because these terminals mediate vasovagal syncope, these data could suggest a mechanistic role of extracellular ATP in this syndrome and, in addition, give further support to the hypothesis that endogenous ATP released from ischemic myocytes is a mediator of atropine-sensitive bradyarrhythmias associated with left ventricular myocardial infarction. PMID- 15539615 TI - Oxidative stress and vascular disease. AB - Growing evidence indicates that chronic and acute overproduction of reactive oxygen species (ROS) under pathophysiologic conditions is integral in the development of cardiovascular diseases (CVD). These ROS can be released from nicotinamide adenine dinucleotide (phosphate) oxidase, xanthine oxidase, lipoxygenase, mitochondria, or the uncoupling of nitric oxide synthase in vascular cells. ROS mediate various signaling pathways that underlie vascular inflammation in atherogenesis: from the initiation of fatty streak development through lesion progress to ultimate plaque rupture. Various animal models of oxidative stress support the notion that ROS have a causal role in atherosclerosis and other cardiovascular diseases. Human investigations also support the oxidative stress hypothesis of atherosclerosis. Oxidative stress is the unifying mechanism for many CVD risk factors, which additionally supports its central role in CVD. Despite the demonstrated role of antioxidants in cellular and animal studies, the ineffectiveness of antioxidants in reducing cardiovascular death and morbidity in clinical trials has led many investigators to question the importance of oxidative stress in human atherosclerosis. Others have argued that the prime factor for the mixed outcomes from using antioxidants to prevent CVD may be the lack of specific and sensitive biomarkers by which to assess the oxidative stress phenotypes underlying CVD. A better understanding of the complexity of cellular redox reactions, development of a new class of antioxidants targeted to specific subcellular locales, and the phenotype-genotype linkage analysis for oxidative stress will likely be avenues for future research in this area as we move toward the broader use of pharmacological and regenerative therapies in the treatment and prevention of CVD. PMID- 15539616 TI - Differential additive effects of endothelial lipase and scavenger receptor-class B type I on high-density lipoprotein metabolism in knockout mouse models. AB - OBJECTIVE: Endothelial lipase (EL) is a vascular phospholipase that hydrolyzes high-density lipoprotein (HDL) as its preferred substrate. Scavenger receptor class B type I (SR-BI) is an HDL receptor that mediates the selective uptake of cholesteryl ester. This study investigates the role of EL and SR-BI in the regulation of HDL metabolism in gene knockout mouse models. METHODS AND RESULTS: We cross-bred EL-/- and SR-BI-/- mice and generated single- and double-null mice. We used biochemical, molecular biology, and nuclear magnetic resonance methods to analyze HDL concentration, composition, and structure. We found that EL and SR-BI display additive effects on HDL with evident gene dosage effects, but their mechanisms to regulate HDL concentration and composition are different. Whereas the elevated HDL cholesterol level in EL-/- mice is associated with increased phospholipid content in HDL particles, SR-BI-/- mice display markedly enlarged HDL particles shifted to larger subclasses with a phospholipid content similar to that of wild-type mice. Furthermore, absence of EL is associated with a 40% to 50% inhibition and absence of SR-BI, a approximately 90% inhibition of endogenous lecithin cholesterol:acyltransferase rate. CONCLUSIONS: EL and SR-BI are major genetic determinants of HDL metabolism in vivo, each exercising independent and additive effects on HDL structure and function. PMID- 15539618 TI - Enhanced cellular adenosine uptake limits adenosine receptor stimulation in patients with hyperhomocysteinemia. AB - OBJECTIVE: Endogenous adenosine has several cardioprotective effects. We postulate that in patients with hyperhomocysteinemia increased intracellular formation of S-adenosylhomocysteine decreases free intracellular adenosine. Subsequently, facilitated diffusion of extracellular adenosine into cells through dipyridamole-sensitive transporters is enhanced, limiting adenosine receptor stimulation. We tested this hypothesis in patients with classical homocystinuria (n=9, plasma homocysteine 93.1+/-24.7 micromol/L) and matched controls (n=8, homocysteine 9.1+/-1.0). METHODS AND RESULTS: Infusion of adenosine (0.5, 1.5, 5.0, and 15.0 microg/min/dL forearm) into the brachial artery increased forearm blood flow, as measured with venous occlusion plethysmography, to 2.9+/-0.4, 4.3+/-0.5, 5.6+/-1.1, and 9.6+/-2.1 in the patients and to 2.8+/-0.6, 4.4+/-1.0, 9.0+/-1.7, and 17.0+/-3.1 mL/min/dL in controls (P<0.05). However, adenosine induced vasodilation in the presence of dipyridamole (100 microg/min/dL) was similar in both groups (P=0.9). Additionally, in isolated erythrocytes, adenosine uptake was accelerated by incubation with homocysteine (half-time 6.4+/-0.3 versus 8.1+/-0.5 minutes, P<0.001) associated with increased intracellular formation of S-adenosylhomocysteine (P<0.0001). CONCLUSIONS: In hyperhomocysteinemia, adenosine-induced vasodilation is impaired but is restored by dipyridamole. Accelerated cellular adenosine uptake probably accounts for these observations. These impaired actions of adenosine could well contribute to the cardiovascular complications of hyperhomocysteinemia. PMID- 15539617 TI - Regulation of transport of the angiotensin AT2 receptor by a novel membrane associated Golgi protein. AB - OBJECTIVE: Synthesis and maturation of G protein-coupled receptors are complex events that require an intricate combination of processes including protein folding, posttranslational modifications, and transport through distinct cellular compartments. Little is known concerning the regulation of G protein-coupled receptor transport from the endoplasmic reticulum to the cell surface. METHODS AND RESULTS: Here we show that the cytoplasmatic carboxy-terminal of the angiotensin AT2 receptor (AT2R) acts independently as an endoplasmic reticulum export signal. Using a yeast two-hybrid system, we identified a Golgi membrane associated protein termed ATBP50 (for AT2R binding protein of 50 kDa) that binds to this motif. We also cloned ATBP60 and ATBP135 encoded by the same gene as ATBP50 that mapped to chromosomes 8p21.3. Downregulation of ATBP50 using siRNA leads to retention of AT2R in inner compartments, reduced cell surface expression, and decreased antiproliferative effects of the receptor. CONCLUSIONS: Our results indicate that ATBP50 regulates the transport of the AT2R to cell membrane by binding to a specific motif within its cytoplasmic carboxy-terminal and thereby enabling the antiproliferative effects of the receptor. PMID- 15539619 TI - Loss of redox factor 1 decreases NF-kappaB activity and increases susceptibility of endothelial cells to apoptosis. AB - OBJECTIVE: The aim of this project was to test the hypothesis that redox factor 1 (Ref-1) was a critical upstream determinant of NF-kappaB-dependent survival signaling pathways in the vessel wall. METHODS AND RESULTS: Aortas from hemizygous transgenic mice harboring a single allele of Ref-1 exhibited a significant loss in NF-kappaB DNA binding activity. The NF-kappaB-dependent survival gene A20 was significantly downregulated in aortas of hemizygous Ref-1 mice, whereas IAP-2 was unchanged. Overexpression of A20 rescued cells from tumor necrosis factor (TNF)-induced apoptosis, suggesting that the loss of A20 in Ref-1 hemizygotes may be a rate-determining step in endothelial cell fate. Deletion of the previously defined redox-sensitive or the AP endonuclease domains of Ref-1 significantly decreased NF-kappaB transcriptional activation and endothelial cell survival. Furthermore, TNF-induced apoptosis was significantly potentiated in endothelial cells after delivery of Morpholino antisense oligodeoxynucleotides targeted to Ref-1. Loss of the redox-sensitive domain blocked the ability of Ref 1 to reduce p50; however, loss of the endonuclease domain did not effect p50 reduction, suggesting alternative mechanisms of action of Ref-1 on NF-kappaB activity. CONCLUSIONS: These findings establish a role for Ref-1 as an upstream determinant of NF-kappaB and A20-dependent signaling and endothelial survival in the vessel wall. PMID- 15539620 TI - Decreased binding of annexin v to endothelial cells: a potential mechanism in atherothrombosis of patients with systemic lupus erythematosus. AB - OBJECTIVE: The cause of the exceedingly high risk of atherothrombosis in systemic lupus erythematosus (SLE) is not clear but antiphospholipid antibodies (aPL) and potentially antithrombotic annexin V have been implicated. METHODS AND RESULTS: Twenty-six women (52+/-8.2 years) with SLE and a history of cardiovascular disease (CVD) (SLE cases) were compared with 26 women with SLE but no CVD (SLE controls) and 26 healthy women (population controls). Common carotid intima-media thickness (IMT) was determined by B-mode ultrasound as a surrogate measure of atherosclerosis. Annexin V binding to human umbilical vein endothelial cells (HUVECs) as determined by flow cytometry after 24-hour culture with plasma was decreased when plasma from SLE cases was used (SLE cases versus population controls: P=0.002; SLE cases versus SLE controls P=0.02). Antibodies against cardiolipin were among IgG antibodies causing decreased binding. There was a positive association between annexin V binding and IMT (R=0.73; P<0.001) among SLE cases. Immunohistochemical analysis revealed presence of annexin V in all human atherosclerotic plaques tested, especially at sites prone to rupture. CONCLUSIONS: Decreased annexin V binding to endothelium caused by antibodies may represent a novel mechanism of atherothrombosis. We hypothesize that even though annexin V may promote plaque growth at some disease stages, it may also stabilize plaque. PMID- 15539621 TI - Role of ADAMTS-1 in atherosclerosis: remodeling of carotid artery, immunohistochemistry, and proteolysis of versican. AB - OBJECTIVE: We investigated the potential role of ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motif type I) in atherogenesis. METHODS AND RESULTS: ADAMTS-1 is expressed at the highest levels in the aorta when compared with other human tissues examined. Immunolocalization studies in human aorta and coronary artery indicate that ADAMTS-1 expression is mainly seen at low levels in the medial layer, but upregulated in the intima when plaque is present. We found that ADAMTS-1 mRNA levels are significantly higher in proliferating/migrating cultured primary aortic vascular smooth muscle cells (VSMCs) compared with resting/confluent cells. Using the mouse carotid artery flow cessation model, we show that there are differences in vessel remodeling in ADAMTS-1 transgenic/apoE deficient mice compared with apoE deficiency alone, particularly a significant increase in intimal hyperplasia. We show that ADAMTS-1 can cleave the large versican containing proteoglycan population purified from cultured human aortic VSMCs. Finally, using versican peptide substrates, we show data suggesting that ADAMTS-1 cleaves versican at multiple sites. CONCLUSIONS: We hypothesize that ADAMTS-1 may promote atherogenesis by cleaving extracellular matrix proteins such as versican and promoting VSMC migration. PMID- 15539622 TI - Macrophage liver X receptor is required for antiatherogenic activity of LXR agonists. AB - OBJECTIVE: Complications of atherosclerotic cardiovascular disease due to elevated blood cholesterol levels are the major cause of death in the Western world. The liver X receptors, LXRalpha and LXRbeta (LXRs), are ligand-dependent transcription factors that act as cholesterol sensors and coordinately control transcription of genes involved in cholesterol and lipid homeostasis as well as macrophage inflammatory gene expression. LXRs regulate cholesterol balance through activation of ATP-binding cassette transporters that promote cholesterol transport and excretion from the liver, intestine, and macrophage. Although LXR agonists are known to delay progression of atherosclerosis in mouse models, their ability to abrogate preexisting cardiovascular disease by inducing regression and stabilization of established atherosclerotic lesions has not been addressed. METHODS AND RESULTS: We demonstrate that LXR agonist treatment increases ATP binding cassette transporter expression within preexisting atherosclerotic lesions, resulting in regression of these lesions as well as remodeling from vulnerable to stable lesions and a reduction in macrophage content. Further, using macrophage-selective LXR-deficient mice created by bone marrow transplantation, we provide the first evidence that macrophage LXR expression is necessary for the atheroprotective actions of an LXR agonist. CONCLUSIONS: These data substantiate that drugs targeting macrophage LXR activity may offer therapeutic benefit in the treatment of atherosclerotic cardiovascular disease. PMID- 15539623 TI - Geographic origin as a determinant of carotid artery intima-media thickness and brachial artery flow-mediated dilation: the Cardiovascular Risk in Young Finns study. AB - OBJECTIVE: People living in eastern Finland have approximately 40% higher coronary heart disease mortality rates than western Finns. Whether this is because of genetic or environmental factors is unknown. We examined the effect of geographic family origin on subclinical atherosclerosis among young Finns. METHODS AND RESULTS: As part of a longitudinal follow-up study, we measured carotid intima-media thickness (IMT) in 2264 and brachial flow-mediated dilation (FMD) in 2109 white adults, aged 24 to 39 years. Subjects from eastern Finland had greater IMT and lower FMD compared with western subjects. These differences accentuated when the subjects' family origin (grandparents' birthplace) was taken into account and remained significant after adjusting for several environmental factors. Among subjects with all grandparents born in eastern or western Finland, IMTs were (mean+/-SEM) 0.592+/-0.003 versus 0.565+/-0.005 mm (P<0.0001), respectively. The corresponding FMD values were 7.61+/-0.15% versus 8.75+/-0.26%; P<0.01. The number of grandparents born in eastern Finland was directly related to IMT (P<0.0001) and inversely to FMD (P<0.05). CONCLUSIONS: Young adults originating from eastern Finland have greater carotid IMT and lower brachial FMD than western Finns. Consistent with a hereditable component predisposing to or protecting from atherosclerosis, these differences accentuated when subjects' family origin was taken into account. We studied whether an east-west difference exists in markers of subclinical atherosclerosis in 2264 Finns aged 24 to 39 years. Subjects with family origin in eastern Finland had greater carotid IMT and lower brachial FMD compared with western subjects, suggesting that hereditable factors play a role in excess atherosclerosis risk in eastern Finland. PMID- 15539624 TI - Protein kinase C pathway is involved in transcriptional regulation of C-reactive protein synthesis in human hepatocytes. AB - OBJECTIVE: C-reactive protein (CRP) is the prototype acute phase protein and a cardiovascular risk factor. Interleukin-1beta (IL-1beta) and IL-6 stimulate CRP synthesis in hepatocytes. We searched for additional pathways regulating CRP expression. METHODS AND RESULTS: Primary human hepatocytes (PHHs) were treated with IL-1beta, IL-6, and protein kinase C (PKC) activator phorbol 12,13 dibutyrate (PDBu). CRP was analyzed by quantitative RT-PCR and ELISA. PDBu significantly induced CRP transcription by 21.0+/-9.24-fold and protein release by 2.9+/-0.5-fold. Transcriptional regulation was studied in detail in hepatoma G2 (HepG2) cells stably transfected with the 1-kb CRP promoter (HepG2-ABEK14 cells). In these cells, PDBu significantly induced CRP transcription by 5.39+/ 0.66-fold. Competitive inhibition with bisindolylmaleimide derivative LY333531 abolished PDBu-mediated promoter activation. Competitive inhibition with IkappaB kinase inhibitor I229 also inhibited PDBu effects. Importantly, IL-8 significantly induced CRP release in PHHs by 58.675+/-19.1-fold, which was blockable by LY333531. CONCLUSIONS: This study describes a novel PKC-dependent transcriptional regulation of CRP gene expression, which, in analogy to the classical IL-1beta and IL-6 pathways, is operational in hepatocytes only. It also identifies IL-8 as a potential physiological PKC activator. HepG2-ABEK14 cells may be useful for high throughput screening to identify inhibitors of CRP synthesis for the prevention of cardiovascular disease. PMID- 15539625 TI - Mercury, fish oils, and risk of acute coronary events and cardiovascular disease, coronary heart disease, and all-cause mortality in men in eastern Finland. AB - OBJECTIVE: Mercury has been suggested to have negative effects on cardiovascular health. We investigated the effects of high mercury content in hair on the risk of acute coronary events and cardiovascular and all-cause mortality in men from eastern Finland. METHODS AND RESULTS: The population-based prospective Kuopio Ischaemic Heart Disease Risk Factor Study (KIHD) cohort of 1871 Finnish men aged 42 to 60 years and free of previous coronary heart disease (CHD) or stroke at baseline was used. During an average follow-up time of 13.9 years, 282 acute coronary events and 132 cardiovascular disease (CVD), 91 CHD, and 525 all-cause deaths occurred. Men in the highest third of hair mercury content (>2.03 microg/g) had an adjusted 1.60-fold (95% CI, 1.24 to 2.06) risk of acute coronary event, 1.68-fold (95% CI, 1.15 to 2.44) risk of CVD, 1.56-fold (95% CI, 0.99 to 2.46) risk of CHD, and 1.38-fold (95% CI, 1.15 to 1.66) risk of any death compared with men in the lower two thirds. High mercury content in hair also attenuated the protective effects of high-serum docosahexaenoic acid plus docosapentaenoic acid concentration. CONCLUSIONS: High content of mercury in hair may be a risk factor for acute coronary events and CVD, CHD, and all-cause mortality in middle-aged eastern Finnish men. Mercury may also attenuate the protective effects of fish on cardiovascular health. PMID- 15539626 TI - Polymorphisms of the interleukin-1beta gene affect the risk of myocardial infarction and ischemic stroke at young age and the response of mononuclear cells to stimulation in vitro. AB - OBJECTIVE: To investigate the role of interleukin-1beta (IL-1beta) gene polymorphisms as a link between inflammation, coagulation, and risk of ischemic vascular disease at young age. METHODS AND RESULTS: A total of 406 patients with myocardial infarction (MI) at young age, frequency-matched for age, sex, and recruitment center, with 419 healthy population-based controls and 134 patients with ischemic stroke at young age, matched by age and sex, with 134 healthy population-based controls, were studied. Subjects carrying the TT genotype of the -511C/T IL-1beta polymorphism showed a decreased risk of MI (odds ratio [OR], 0.36; 95% CI, 0.20 to 0.64) and stroke (OR, 0.32; 95% CI, 0.13 to 0.81) after adjustment for conventional risk factors. In both studies, the T allele showed a codominant effect (P=0.0020 in MI; P=0.021 in stroke). Mononuclear cells from volunteers carrying the T allele showed a decreased release of IL-1beta and a decreased expression of tissue factor after stimulation with lipopolysaccharide compared with CC homozygotes. The presence of a monoclonal antibody against IL 1beta during cell stimulation resulted in a marked reduction of tissue factor activity expression. CONCLUSIONS: -511C/T IL-1beta gene polymorphism affects the risk of MI and ischemic stroke at young age and the response of mononuclear cells to inflammatory stimulation. PMID- 15539630 TI - Peroxisome proliferator-activated receptor alpha induces NADPH oxidase activity in macrophages, leading to the generation of LDL with PPAR-alpha activation properties. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors controlling lipid and glucose metabolism as well as inflammation. PPARs are expressed in macrophages, cells that also generate reactive oxygen species (ROS). In this study, we investigated whether PPARs regulate ROS production in macrophages. Different PPAR-alpha, but not PPAR-gamma agonists, increased the production of ROS (H2O2 and ) in human and murine macrophages. PPAR-alpha activation did not induce cellular toxicity, but significantly decreased intracellular glutathione levels. The increase in ROS production was not attributable to inherent prooxidant effects of the PPAR-alpha agonists tested, but was mediated by PPAR-alpha, because the effects were lost in bone marrow derived macrophages from PPAR-alpha-/- mice. The PPAR-alpha-induced increase in ROS was attributable to the induction of NADPH oxidase, because (1) preincubation with the NADPH oxidase inhibitor diphenyleneiodinium prevented the increase in ROS production; (2) PPAR-alpha agonists increased production measured by superoxide dismutase-inhibitable cytochrome c reduction; (3) PPAR-alpha agonists induced mRNA levels of the NADPH oxidase subunits p47(phox), p67phox, and gp91phox and membrane p47phox protein levels; and (4) induction of ROS production was abolished in p47phox-/- and gp91phox-/- macrophages. Finally, induction of NADPH oxidase by PPAR-alpha agonists resulted in the formation of oxidized LDL metabolites that exert PPAR-alpha-independent proinflammatory and PPAR-alpha dependent decrease of lipopolysaccharide-induced inducible nitric oxide synthase expression in macrophages. These data identify a novel mechanism of autogeneration of endogenous PPAR-alpha ligands via stimulation of NADPH oxidase activity. PMID- 15539631 TI - Peptidyl-glycine alpha-amidating monooxygenase targeting and shaping of atrial secretory vesicles: inhibition by mutated N-terminal ProANP and PBA. AB - ANP (atrial natriuretic peptide) is widely recognized as an important vasorelaxant, diuretic, and cardioprotective hormone. Little is known, however, about how ANP-secretory vesicles form within the atrial myocytes. Secretory vesicles were visualized by fluorescence microscope imaging in live rat atrial myocytes expressing proANP-enhanced green fluorescent protein (EGFP), or N terminal-mutated fusion proteins thought to suppress the calcium-dependent aggregation of proANP. Results showed the following: (1) aggregates of proANP and coexpressed proANP-EGFP recruited peptidylglycine alpha-amidating monooxygenase (PAM)-1, an abundant atrial integral vesicle membrane protein; (2) coexpressed N terminal-mutated (Glu23,24-->Gln23,24) and N-terminal-deleted proANP-EGFP inhibited recruitment of PAM-1 by up to 60%; (3) 4-phenyl-3-butenoic acid (PBA) (10 mumol/L), a pharmacological inhibitor of the lumenal peptidylglycine alpha hydroxylating monooxygenase domain of PAM proteins, inhibited recruitment of endogenous PAM-1 and of coexpressed pro-EGFP-PAM-1; (4) PBA had no effect on exocytosis of the potassium inward rectifier KIR2.1; (5) PBA induced a deformation of the secretory vesicles but did not inhibit docking. These findings suggest that recruitment of PAM-1 to secretory vesicles depends on intact N terminal proANP and on the lumenal domain of PAM-1. Conversely, PAM-1 participates in shaping the proANP-secretory vesicles. The full text of this article is available online at http://circres.ahajournals.org. PMID- 15539632 TI - Molecular imaging of atherosclerotic plaques using a human antibody against the extra-domain B of fibronectin. AB - Current imaging modalities of human atherosclerosis, such as angiography, ultrasound, and computed tomography, visualize plaque morphology. However, methods that provide insight into plaque biology using molecular tools are still insufficient. The extra-domain B (ED-B) is inserted into the fibronectin molecule by alternative splicing during angiogenesis and tissue remodeling but is virtually undetectable in normal adult tissues. Angiogenesis and tissue repair are also hallmarks of advanced plaques. For imaging atherosclerotic plaques, the human antibody L19 (specific against ED-B) and a negative control antibody were labeled with radioiodine or infrared fluorophores and injected intravenously into atherosclerotic apolipoprotein E-null (ApoE-/-) or normal wild-type mice. Aortas isolated 4 hours, 24 hours, and 3 days after injection exhibited a selective and stable uptake of L19 when using radiographic or fluorescent imaging. L19 binding was confined to the plaques as assessed by fat staining. Comparisons between fat staining and autoradiographies 24 hours after 125I-labeled L19 revealed a significant correlation (r=0.89; P<0.0001). Minimal antibody uptake was observed in normal vessels from wild-type mice receiving the L19 antibody and in atherosclerotic vessels from ApoE-/- mice receiving the negative control antibody. Immunohistochemical studies revealed increased expression of ED-B not only in murine but also in human plaques, in which it was found predominantly around vasa vasorum and plaque matrix. In summary, we demonstrate selective targeting of atheromas in mice using the human antibody to the ED-B domain of fibronectin. Thus, our findings may set the stage for antibody-based molecular imaging of atherosclerotic plaques in the intact organism. PMID- 15539633 TI - Liver X receptor agonists suppress vascular smooth muscle cell proliferation and inhibit neointima formation in balloon-injured rat carotid arteries. AB - The liver X receptors alpha and beta (LXRalpha and LXRbeta) are important regulators of cholesterol homeostasis in liver and macrophages. Synthetic LXR ligands prevent the development of atherosclerosis in murine models; however, the potential functional relevance of LXRs in vascular smooth muscle cells (VSMCs) has not been investigated. In the present study, we demonstrate that LXRs are expressed and functional in primary human coronary artery VSMCs (CASMCs). LXR ligands inhibited mitogen-induced VSMC proliferation and G1-->S phase progression of the cell cycle. Inhibition of G1 exit by LXR ligands was accompanied by a dose dependent inhibition of retinoblastoma protein (Rb) phosphorylation, which functions as the key switch for G1-->S cell cycle progression. LXR ligands suppressed mitogen-induced degradation of the cyclin-dependent kinase inhibitor p27Kip1, attenuated cyclin D1 and cyclin A expression, and inhibited the expression of S phase-regulatory minichromosome maintenance protein 6. Stabilization of p27kip1 by LXR ligands was mediated by supressing the transcriptional activation of the S phase kinase-associated protein 2 (Skp2), an F-box protein that targets p27Kip1 for degradation. Inhibition of Rb phosphorylation and G1-->S cell cycle progression by LXR ligands was reversed in VSMCs overexpressing Skp2, indicating that Skp2 as an upstream regulator of p27Kip1 degradation plays a central role in LXR ligand-mediated inhibition of VSMC proliferation. Furthermore, adenovirus-mediated overexpression of the S phase transcription factor E2F, which is released after Rb phosphorylation, reversed the inhibitory effect of LXR ligands on VSMC proliferation and S phase gene expression, suggesting that the primary mechanisms by which LXR ligands inhibit VSMC proliferation occur upstream of Rb phosphorylation. Finally, neointima formation in a model of rat carotid artery balloon injury was significantly attenuated after treatment with the LXR ligand T1317 compared with vehicle-treated animals. These data demonstrate that LXR ligands inhibit VSMC proliferation and neointima formation after balloon injury and suggest that LXR ligands may constitute a novel therapy for proliferative vascular diseases. The full text of this article is available online at http://circres.ahajournals.org. PMID- 15539634 TI - Transforming growth factor beta receptor endoglin is expressed in cardiac fibroblasts and modulates profibrogenic actions of angiotensin II. AB - Angiotensin II (Ang II) is a powerful mediator of adverse cardiac remodeling and fibrosis. However, the mechanisms of Ang II-induced myocardial fibrosis remain to be clarified. We postulated that Ang II alters transforming growth factor beta (TGF-beta) receptor expression, specifically that of endoglin, and thereby modulates cardiac fibroblast (CF) collagen metabolism. Experiments were conducted using CF from adult Sprague Dawley rats to determine the expression of TGF-beta1 receptors including endoglin, and the role of Ang II type 1 (AT1) and type 2 (AT2) receptors, and MAPK p42/44 in this process. The functional role of endoglin in modulating Ang II effects on matrix metalloproteinase-1 (MMP-1) and type I collagen expression was also analyzed. Endoglin gene and protein expression were consistently identified in quiescent CFs. Ang II increased the expression of endoglin mRNA and protein in a concentration and time-dependent manner, with no effect on TGF-beta receptors I and II expression. This effect was AT1 receptor mediated, because AT1 receptor antagonists valsartan, candesartan, and losartan inhibited Ang II-induced endoglin expression, whereas the AT2 receptor antagonist PD123319 had no effect. MAPKp42/44 inhibition attenuated Ang II-induced endoglin expression. Ang II-induced decrease in MMP-1 protein expression and increase in type I collagen protein expression were both blocked by a specific endoglin antibody. Hence, our results indicate that endoglin is upregulated in CFs by Ang II via the AT1 receptor and modulates profibrotic effects of Ang II. These findings provide novel insights into Ang II-induced cardiac remodeling. PMID- 15539635 TI - Physiological growth synergizes with pathological genes in experimental cardiomyopathy. AB - Hundreds of signaling molecules have been assigned critical roles in the pathogenesis of myocardial hypertrophy and heart failure based on cardiac phenotypes from alpha-myosin heavy chain-directed overexpression mice. Because permanent ventricular transgene expression in this system begins during a period of rapid physiological neonatal growth, resulting phenotypes are the combined consequences of transgene effects and normal trophic influences. We used temporally-defined forced gene expression to investigate synergy between postnatal physiological cardiac growth and two functionally divergent cardiomyopathic genes. Phenotype development was compared various times after neonatal (age 2 to 3 days) and adult (age 8 weeks) expression. Proapoptotic Nix caused ventricular dilation and severe contractile depression in neonates, but not adults. Myocardial apoptosis was minimal in adults, but was widespread in neonates, until it spontaneously resolved in adulthood. Unlike normal postnatal cardiac growth, concurrent left ventricular pressure overload hypertrophy did not synergize with Nix expression to cause cardiomyopathy or myocardial apoptosis. Prohypertrophic Galphaq likewise caused eccentric hypertrophy, systolic dysfunction, and pathological gene expression in neonates, but not adults. Thus, normal postnatal cardiac growth can be an essential cofactor in development of genetic cardiomyopathies, and may confound the interpretation of conventional alpha-MHC transgenic phenotypes. PMID- 15539636 TI - Phosphatidylinositol 3-kinase offsets cAMP-mediated positive inotropic effect via inhibiting Ca2+ influx in cardiomyocytes. AB - Phosphoinositide 3-kinase (PI3K) has been implicated in beta2-adrenergic receptor (beta2-AR)/G(i)-mediated compartmentation of the concurrent G(s)-cAMP signaling, negating beta2-AR-induced phospholamban phosphorylation and the positive inotropic and lusitropic responses in cardiomyocytes. However, it is unclear whether PI3K crosstalks with the beta1-AR signal transduction, and even more generally, with the cAMP/PKA pathway. In this study, we show that selective beta1 AR stimulation markedly increases PI3K activity in adult rat cardiomyocytes. Inhibition of PI3K by LY294002 significantly enhances beta1-AR-induced increases in L-type Ca2+ currents, intracellular Ca2+ transients, and myocyte contractility, without altering the receptor-mediated phosphorylation of phospholamban. The LY294002 potentiating effects are completely prevented by betaARK-ct, a peptide inhibitor of beta-adrenergic receptor kinase-1 (betaARK1) as well as G(betagamma) signaling, but not by disrupting G(i) function with pertussis toxin. Moreover, forskolin, an adenylyl cyclase activator, also elevates PI3K activity and inhibition of PI3K enhances forskolin-induced contractile response in a betaARK-ct sensitive manner. In contrast, PI3K inhibition affects neither the basal contractility nor high extracellular Ca2+ induced increase in myocyte contraction. These results suggest that beta1-AR stimulation activates PI3K via a PKA-dependent mechanism, and that G(betagamma) and the subsequent activation of betaARK1 are critically involved in the PKA induced PI3K signaling which, in turn, negates cAMP-induced positive inotropic effect via inhibiting sarcolemmal Ca2+ influx and the subsequent increase in intracellular Ca2+ transients, without altering the receptor-mediated phospholamban phosphorylation, in intact cardiomyocytes. PMID- 15539637 TI - All arteriogenesis is local? Home boys versus the newcomers. PMID- 15539638 TI - Sir-viving cardiac stress: cardioprotection mediated by a longevity gene. PMID- 15539639 TI - The mitochondrial death pathway and cardiac myocyte apoptosis. AB - Apoptosis has been causally linked to the pathogenesis of myocardial infarction and heart failure in rodent models. This death process is mediated by two central pathways, an extrinsic pathway involving cell surface receptors and an intrinsic pathway using mitochondria and the endoplasmic reticulum. Each of these pathways has been implicated in myocardial pathology. In this review, we summarize recent advances in the understanding of the intrinsic pathway and how it relates to cardiac myocyte death and heart disease. PMID- 15539640 TI - Expression of deoxynucleotide carrier is not associated with the mitochondrial DNA depletion caused by anti-HIV dideoxynucleoside analogs and mitochondrial dNTP uptake. AB - Our previous studies suggested that the dNTP/dNDP transporter systems that exist in mitochondria for transporting dNTP/dNDP from the cytoplasm to the mitochondria for mitochondrial DNA (mtDNA) synthesis play a critical role in delayed cytotoxicity of anti-human immunodeficiency virus (HIV) dideoxynucleoside analogs in mitochondria. A protein, termed mitochondrial deoxynucleotide carrier (DNC), based on its ability to transport dNTPs in reconstituted proteoliposomes, was recently isolated. Lacking cellular information to substantiate DNC's involvement in the delayed cytotoxicity of dideoxynucleoside analogs, we expressed DNC and reconstituted it into proteoliposomes. The K(m) values for dNTPs uptake by reconstituted DNC were in the millimolar range, which is a thousandfold higher than that of the physiological level. Furthermore, we found that overexpressing DNC (wt and G177A-mutated DNC) in RKO cells did not sensitize the cells to the mtDNA depletion caused by beta-d-2',3'-dideoxycytidine (ddC), 2',3'-didehydro 2',3'-dideoxythymidine, and 2',3'-dideoxyinosine or affect the mtDNA recovery rate after ddC treatment. Mitochondria isolated from DNC-overexpressing cells did not significantly differ from that isolated from RKO cells in terms of the rate of uptake or the incorporation of dTTP into mitochondria DNA. Down-regulation of DNC expression by small interfering RNA was also ineffective in changing the action of dideoxynucleoside analogs on the mtDNA depletion and the rate of dTTP uptake into isolated mitochondria. Down-regulation of both DNC and thymidine kinase-2 also did not cause mtDNA depletion. We conclude that DNC does not play an important role in the delayed cytotoxicity (mtDNA depletion) of anti-HIV dideoxynucleoside analogs and dNTPs uptake into mitochondria. PMID- 15539641 TI - Adenosine A2A receptor and dopamine D3 receptor interactions: evidence of functional A2A/D3 heteromeric complexes. AB - Adenosine A(2A) and dopamine D(2) receptors have been shown previously to form heteromeric complexes and interact at the level of agonist binding, G protein coupling, and trafficking. Because dopamine D(2) and D(3) receptors show a high degree of sequence homology, A(2A) and D(3) receptors may also interact in a similar manner. The present studies with confocal microscopy showed that A(2A) yellow fluorescent protein (YFP) and D(3)-green fluorescent protein 2 (GFP2) receptors colocalize in the plasma membrane. Furthermore, fluorescence resonance energy transfer (FRET) analysis demonstrated that A(2A)-YFP and D(3)-GFP2 receptors give a positive FRET efficiency and are thereby likely to exist as heteromeric A(2A)/D(3) receptor complexes. Saturation experiments with [(3)H]dopamine demonstrated that the A(2A) receptor agonist 4-[2-[[6-amino-9(N ethyl-beta-d-ribofuranuronaminoamidosyl)-9H-purin-2 yl]amino]ethyl]benzenepropanoic acid (CGS-21680) reduced the affinity of the high affinity agonist binding state of the D(3) receptor for [(3)H]dopamine. The A(2A) and D(2A) receptors seem to interact also at the level of G protein coupling, because the adenosine A(2A) receptor agonist CGS-21680 fully counteracted the D(3) receptor-mediated inhibition of a forskolin-mediated increase in cAMP levels. Taken together, when coexpressed in the same neuron, A(2A) and D(3) receptors seem to form A(2A)/D(3) heteromeric receptor complexes in which A(2A) receptors antagonistically modulate both the affinity and the signaling of the D(3) receptors. D(3) receptor is one of the therapeutic targets for treatment of schizophrenia, and therefore, the A(2A)/D(3) receptor interactions could provide an alternative antischizophrenic treatment. PMID- 15539642 TI - Poor countries make the best teachers: discuss. PMID- 15539643 TI - Standards of care in research. PMID- 15539644 TI - Lessons from thalassaemia screening in Iran. PMID- 15539645 TI - Health systems and the community. PMID- 15539646 TI - Highly active antiretroviral therapy. PMID- 15539647 TI - Knowledge for better health. PMID- 15539648 TI - WHO denies suppressing report on junk food. PMID- 15539649 TI - European Union may relax rules on marketing baby milk. PMID- 15539652 TI - Will Bush's insurance scheme help the uninsured? PMID- 15539658 TI - Supporting surgery for obstetric fistula. PMID- 15539659 TI - An extra dollar can go a long way. PMID- 15539660 TI - Demystifying health care. PMID- 15539661 TI - Babies fed defective formula are still being treated for neurological damage. PMID- 15539664 TI - From looking at cells to looking at systems. PMID- 15539666 TI - Iranian national thalassaemia screening programme. PMID- 15539665 TI - Crotaline snake bite in the Ecuadorian Amazon: randomised double blind comparative trial of three South American polyspecific antivenoms. AB - OBJECTIVE: To compare the efficacy and safety of three polyspecific antivenoms for bites by pit vipers. DESIGN: Randomised double blind comparative trial of three antivenoms. SETTING: Shell, Pastaza, southeastern Ecuador. PARTICIPANTS: 210 patients with incoagulable blood were recruited from 221 consecutive patients admitted with snake bite between January 1997 and December 2001. INTERVENTION: One of three antivenoms manufactured in Brazil, Colombia, and Ecuador, chosen for their preclinical potency against Ecuadorian venoms. MAIN OUTCOME MEASURES: Permanent restoration of blood coagulability after 6 and 24 hours. RESULTS: The snakes responsible for the bites were identified in 187 cases: 109 patients (58%) were bitten by Bothrops atrox, 68 (36%) by B bilineatus, and 10 (5%) by B taeniatus, B brazili, or Lachesis muta. Eighty seven patients (41%) received Colombian antivenom, 82 (39%) received Brazilian antivenom, but only 41 (20%) received Ecuadorian antivenom because the supply was exhausted. Two patients died, and 10 developed local necrosis. All antivenoms achieved the primary end point of permanently restoring blood coagulability by 6 or 24 hours after the start of treatment in > 40% of patients. Colombian antivenom, however, was the most effective after initial doses of 20 ml (two vials), < 70 ml, and any initial dose at both 6 and 24 hours. An initial dose of 20 ml of Colombian antivenom permanently restored blood coagulability in 64% (46/72) of patients after 6 hours (P = 0.054 compared with the other two antivenoms) and an initial dose of < 70 ml was effective at 6 hours (65%, P = 0.045) and 24 hours (99%, P = 0.06). Early anaphylactoid reactions were common (53%, 73%, and 19%, respectively, for Brazilian, Colombian, and Ecuadorian antivenoms, P < 0.0001) but only three reactions were severe and none was fatal. CONCLUSIONS: All three antivenoms can be recommended for the treatment of snakebites in this region, though the reactogenicity of Brazilian and Colombian antivenoms is a cause for concern. PMID- 15539667 TI - Learning from low income countries: mental health. PMID- 15539668 TI - Community models of mental care warrant more governmental support. PMID- 15539669 TI - An information system and medical record to support HIV treatment in rural Haiti. PMID- 15539670 TI - Prevention of mother to child transmission of HIV: evaluation of a pilot programme in a district hospital in rural Zimbabwe. AB - PROBLEM: Zimbabwe has one of the highest rates of HIV seroprevalence in the world. In 2001 only 4% of women and children in need of services for prevention of mother to child transmission of HIV were receiving them. DESIGN: Pilot implementation of the first programme for prevention of mother to child transmission of HIV in rural Zimbabwe. SETTING: 120 bed district hospital in Buhera district (285,000 inhabitants), Manicaland, Zimbabwe. KEY MEASURES FOR IMPROVEMENT: Programme uptake indicators monitored for 18 months; impact of policy evaluated by assessing up-scaling of programme. STRATEGIES FOR CHANGE: Voluntary counselling and testing services for HIV were provided in the hospital antenatal clinic. Women identified as HIV positive and informed of their serostatus and their newborn were offered a single dose antiretroviral treatment of nevirapine; mother-child pairs were followed up through routine health services. Nursing staff and social workers were trained, and community mobilisation was conducted. EFFECTS OF CHANGE: No services for prevention of mother to child transmission of HIV were available at baseline. Within 18 months, 2298 pregnant women had received pretest counselling, and the acceptance of HIV testing reached 93.0%. Of all 2137 women who had an HIV test, 1588 (74.3%) returned to collect their result; 326 of the 437 HIV positive women diagnosed had post-test counselling, and 104 (24%) mother-child pairs received nevirapine prophylaxis. LESSONS LEARNT: Minimum staffing, an enhanced training programme, and involvement of district health authorities are needed for the implementation and successful integration of services for prevention of mother to child transmission of HIV. Voluntary counselling and testing services are important entry points for HIV prevention and care and for referral to community networks and medical HIV care services. A district approach is critical to extend programmes for prevention of mother to child transmission of HIV in rural settings. The lessons learnt from this pilot programme have contributed to the design of the national expansion strategy for prevention of mother to child transmission of HIV in Zimbabwe. PMID- 15539671 TI - Reducing length of stay in hospital for very low birthweight infants by involving mothers in a stepdown unit: an experience from Karachi (Pakistan). AB - PROBLEM: Clinical care of infants with a very low birth weight (less than 1500 g) in developing countries can be labour intensive and is often associated with a prolonged stay in hospital. The Aga Khan University Medical Center in Karachi, Pakistan, established a neonatal intensive care unit in 1987. By 1993-4, very low birthweight infants remained in hospital for 18-21 days. STRATEGIES FOR CHANGE: A stepdown unit was established in September 1994, with mothers providing all basic nursing care for their infants before being discharged under supervision. KEY MEASURES FOR IMPROVEMENT: We analysed neonatal outcomes for the time periods before and after the stepdown unit was created (1987-94 and 1995-2001). We compared these two time periods for survival after birth until discharge, morbidity patterns during hospitalisation, length of stay in hospital, and readmission rates to hospital in the four weeks after discharge. EFFECTS OF CHANGE: Of 509 consecutive, very low birthweight infants, 494 (97%) preterm and 140 (28%) weighing < 1000 g at birth), 391 (76%) survived to discharge from the hospital. The length of hospitalisation fell significantly from 1987-90, when it was 34 (SD 18) days, to 16 (SD 14) days in 1999-2001 (P < 0.001). Readmission rates to hospital did not rise, nor did adverse outcomes at 12 months of age. LESSONS LEARNT: Our results indicate that it is possible to involve mothers in the active care of their very low birthweight infants before discharge. This may translate into earlier discharge from hospital to home settings without any increase in short term complications and readmissions. PMID- 15539672 TI - Traditional herbal medicines for malaria. PMID- 15539673 TI - Innovative low cost technologies for biomedical research and diagnosis in developing countries. PMID- 15539674 TI - Expanding antiretroviral therapy in Malawi: drawing on the country's experience with tuberculosis. PMID- 15539675 TI - Reducing maternal and neonatal mortality in the poorest communities. PMID- 15539676 TI - The concept of essential medicines: lessons for rich countries. PMID- 15539677 TI - The pros and cons of essential medicines for rich countries. PMID- 15539678 TI - Learning from failed health reform in Uganda. PMID- 15539679 TI - The devil is in the detail. PMID- 15539680 TI - The power of positive deviance. PMID- 15539681 TI - Kangaroo Mother Care, an example to follow from developing countries. PMID- 15539682 TI - Family friendly care. PMID- 15539683 TI - Learning from low income countries: what are the lessons? Community oral rehydration units can contain cholera epidemics. PMID- 15539684 TI - Learning from low income countries: what are the lessons? Communities improve health systems. PMID- 15539685 TI - Learning from low income countries: what are the lessons? Communities should decide priorities. PMID- 15539686 TI - Learning from low income countries: what are the lessons? Partnerships in mental health are possible without multidisciplinary teams. PMID- 15539687 TI - Learning from low income countries: what are the lessons?: Hands on course may help deliver obstetric care. PMID- 15539688 TI - Learning from low income countries: what are the lessons? Trained medical assistants can successfully do work of doctors. PMID- 15539689 TI - Learning from low income countries: what are the lessons?: Palliative care can be delivered through neighbourhood networks. PMID- 15539690 TI - Learning from low income countries: what are the lessons? Health is a dynamic process. PMID- 15539691 TI - Learning from low income countries: what are the lessons? Emphasis on sophisticated investigations could be reduced, for example. PMID- 15539692 TI - Learning from low income countries: what are the lessons? Health information technology need not cost the earth. PMID- 15539693 TI - Learning from low income countries: what are the lessons? Effective surgery can be cheap and innovative. PMID- 15539694 TI - Learning from low income countries: what are the lessons? Sources of information should focus on developing countries. PMID- 15539695 TI - Learning from low income countries: what are the lessons? Website highlights ethics issues for research in developing countries. PMID- 15539696 TI - Learning from low income countries: what are the lessons? Improvement can occur even in resource constrained settings. PMID- 15539697 TI - Effective management of pericardial neoplasia. PMID- 15539698 TI - Does benign "primary snoring" ever exist in children? PMID- 15539699 TI - Gastroesophageal reflux as an asthma trigger: acid stress. PMID- 15539700 TI - The oximeter: boon or bane? PMID- 15539701 TI - Ventilator-associated pneumonia in institutionalized elders: are teeth a reservoir for respiratory pathogens? PMID- 15539702 TI - ICU care at the end of life. PMID- 15539703 TI - Immunostimulation: does it work in COPD? PMID- 15539704 TI - Preventing medical errors, avoiding litigation: not easy to do in 2004. PMID- 15539705 TI - Decreased recognition of the post-myocardial infarction (Dressler) syndrome in the postinfarct setting: does it masquerade as "idiopathic pericarditis" following silent infarcts? PMID- 15539706 TI - Long-term results of intrapericardial chemotherapeutic treatment of malignant pericardial effusions with thiotepa. AB - OBJECTIVE: Pericardial involvement is a common feature in different neoplastic diseases, having a strong influence on the natural history of the disease and on the quality of life of the patients. This study was performed in order to investigate the long-term effects of intracavitary treatment with thiotepa in the reduction of pericardial effusion (PE) recurrences. DESIGN: Prospective controlled intervention study. SETTING: European Institute of Oncology, Milan, Italy. PATIENTS: We studied 33 patients, 15 men and 18 women, with malignant PE, who were affected by breast cancer (11 patients), lung cancer (16 patients), microcytoma (4 patients), endometrial cancer (1 patients), and melanoma (1 patient). INTERVENTION: All patients with large PE, with or without cardiac tamponade, underwent percutaneous pericardiocentesis (PC) under echocardiographic monitoring. Patients with neoplastic cells in drained fluid were considered to be eligible for treatment. After drainage, the catheter was maintained in the pericardial sac for the instillation of a sclerosing, alkylating antiblastic agent (thiotepa) on days 1, 3, and 5 after the PC (15 mg at each step). RESULTS: No procedure-related complications or side effects were observed. Two patients died because of disease progression, without PE evidence. No PE occurred in the remaining patients during the first month. Three recurrences occurred (9.1%), requiring additional PC and intrapericardial treatment. The median survival time was 115 days (range, 22 to 1,108 days) in the overall population, and 272 days in patients with breast cancer. CONCLUSIONS: Intrapericardial treatment with thiotepa carries a minimal risk and is a repeatable procedure that can dramatically increase quality of life, or even can improve survival and the natural history of disease in cancer patients. PMID- 15539707 TI - Levels and values of serum high-sensitivity C-reactive protein within 6 hours after the onset of acute myocardial infarction. AB - BACKGROUND: C-reactive protein (CRP), which has been suggested to directly enhance inflammation in plaques, is rapidly synthesized and secreted in the liver 6 h after an acute inflammatory stimulus. Therefore, serum levels of CRP within 6 h after the onset of acute myocardial infarction (AMI) merely reflect a chronic and persistent inflammatory process and are not due to acute myocardial damage. We hypothesized that the serum CRP level, which would abnormally elevate thereafter, is followed by a plaque rupture in the clinical setting of AMI. METHODS AND RESULTS: CRP was prospectively measured by high-sensitivity CRP assay (hs-CRP) in 157 consecutive patients (106 patients within 6 h, and 51 patients >/= 6 h but < 12 h after the onset of AMI) with ST-segment elevation AMI undergoing primary percutaneous coronary intervention (PCI). Serum levels of hs CRP were also measured in 30 patients with stable angina undergoing elective PCI and in 30 healthy control subjects. The serum level of hs-CRP was significantly higher in patients with an onset of AMI < 6 h than in patients with angina pectoris (2.7 +/- 2.3 mg/L vs 1.4 +/- 0.7 mg/L, p < 0.0001 [mean +/- SD]) and in healthy subjects (2.7 +/- 2.3 mg/L vs 1.0 +/- 0.6 mg/L, p < 0.0001). There were no significant differences in serum levels of hs-CRP in patients with an onset of AMI 3 h but < 6 h (2.7 +/- 2.5 mg/L vs 2.7 +/- 2.2 mg/L, p = 0.87). However, the serum level of hs-CRP was significantly higher in patients with an onset >/= 6 h than in patients with an onset < 6 h (14.1 +/- 16.5 mg/L vs 2.7 +/- 2.3 mg/L, p < 0.0001). CONCLUSIONS: Serum levels of hs-CRP were significantly higher in patients with an onset of AMI < 6 h than in healthy subjects and in patients with angina pectoris undergoing PCI. The inflammatory process has been proved as one of the mechanisms causing plaque rupture. Elevated serum hs-CRP levels in patients with AMI < 6 h may portend vulnerable plaque rupture. PMID- 15539708 TI - Sarcoidosis diagnosed in elderly subjects: retrospective study of 30 cases. AB - STUDY OBJECTIVE: This study investigated the clinical features and disease course of sarcoidosis diagnosed in patients > 70 years of age. METHODS: A retrospective analysis was made of cases treated at the University Hospital in Nantes, France, between 1986 and 2000. The diagnosis of sarcoidosis was confirmed histopathologically. Cases involving progressive cancer and active tuberculosis were excluded. RESULTS: Thirty white patients with sarcoidosis diagnosed after age 70 years (mean, 74 years) were included. An alteration of general health (asthenia and/or anorexia and/or weight loss) was frequent (53%) and characteristic of the systemic form of the disease. Dyspnea was a fairly common sign (23%). The intrathoracic form of sarcoidosis was most frequent (43.3%). Diagnosis was difficult and lengthy, and symptomatology was atypical. Accessory salivary gland biopsy was an important contributing factor to diagnosis (70.6% were positive). Oral corticosteroid therapy was often required (60.7%). The disease course was satisfactory overall (81.8% of cases), but only for 50% of patients in intrathoracic stage IV. CONCLUSIONS: The clinical presentation of sarcoidosis in elderly subjects is mainly characterized by an alteration of general health. Diagnosis is difficult and should include accessory salivary gland biopsy. Therapy frequently involves corticosteroids. Overall prognosis is similar to that for young subjects. PMID- 15539709 TI - Trends and occupational associations in incidence of hospitalized pulmonary sarcoidosis and other lung diseases in Navy personnel: a 27-year historical prospective study, 1975-2001. AB - STUDY OBJECTIVES: This study examines long-term trends in incidence rates of hospitalized pulmonary sarcoidosis in a large cohort of Navy personnel, and evaluates the possible relationship of sarcoidosis with occupation. DESIGN: Incidence rates of first hospitalizations were determined for black and white male Navy enlisted personnel on active duty from 1975 to 2001. SETTING: Navy service includes a potential for exposure to a variety of substances, including nonskid coatings used on ship decks that may be aerosolized during removal. Particulate matter containing aluminum, titanium, and silicates has been identified in nonskid samples. Specific occupational groups may have had greater exposure potential than others. PATIENTS OR PARTICIPANTS: Hospitalized cases included sarcoidosis (n = 674), asthma (n = 3,536), emphysema and chronic bronchitis (n = 1,103), respiratory conditions due to fumes and vapors (n = 61), and pneumoconiosis (n = 51) observed in 9,953,607 person-years of active-duty service. INTERVENTIONS: None. However, improvements were made in personal protective gear and other countermeasures to prevent or limit respiratory exposures during service. MEASUREMENTS AND RESULTS: Annual overall hospitalized sarcoidosis incidence rates per 100,000 were 24.9 for black men and 3.5 for white men (black/white ratio of 7.1, p < 0.0001). Annual incidence rates in blacks declined markedly, particularly since 1989, but the black/white ratio remained high through 1999. Occupational associations were present in blacks and whites. Black ship's servicemen (23 cases) and aviation structural mechanics specializing in structures (12 cases) had more than twice the expected incidence rate compared to all blacks, and white mess management specialists (15 cases) had twice the overall white incidence rate. CONCLUSIONS: There was a steep decline in incidence of hospitalized sarcoidosis in blacks in the Navy. Occupational associations suggest the possibility that a dust or moisture-related lung disease may have been erroneously classified as sarcoidosis, or, alternatively, that sarcoidosis had a previously unrecognized occupational component. PMID- 15539710 TI - Regional differences in emphysema scores and BAL glutathione levels in HIV infected individuals. AB - STUDY OBJECTIVES: Evidence exists that HIV-seropositive individuals may be at increased risk for the development of precocious pulmonary emphysema. HIV infection is also associated with antioxidant deficiency in both the serum and lungs, and it is therefore possible that increased oxidant stress may contribute to parenchymal lung injury occurring in the setting of HIV. We sought to determine the regional distribution of emphysema and regional distribution of glutathione (GSH) concentrations among HIV-seropositive subjects with emphysema. DESIGN: Cross-sectional evaluation of a prospective, longitudinal study. SETTING: University teaching hospital. SUBJECTS/MEASUREMENTS: HIV-seropositive subjects without AIDS-related pulmonary complications participating in a descriptive study of lung biology in HIV-seropositive individuals. Emphysema scoring and evaluation of emphysema lobar distribution was performed among 40 subjects with emphysema. Eleven subjects underwent BAL of the right middle lobe (RML) and right upper lobe (RUL) with measurement of epithelial lining fluid (ELF) GSH in each lobe. RESULTS: We found that the mean emphysema scores were much higher in the upper lobes compared to the rest of the lung. Mean GSH levels were significantly greater in the RUL compared to the RML. The regional differences were present in both smokers and nonsmokers. CONCLUSIONS: We conclude that in the setting of HIV, emphysema is more prominent and lung GSH concentrations are higher in the upper lobes. We hypothesize that the increased GSH may represent a compensatory response to increased oxidant stress in the upper lobes. PMID- 15539711 TI - Predictors of survival in severe, early onset COPD. AB - STUDY OBJECTIVES: Multiple risk factors for mortality in patients with COPD have been described, but most studies have involved older, primarily male subjects. The purpose of this study was to determine the mortality rate and predictors of survival in subjects with severe, early onset COPD. DESIGN, SETTING, AND PARTICIPANTS: The cohort of 139 probands in the Boston Early-Onset COPD Study was recruited from lung transplant and general pulmonary clinics between September 1994 and July 2002. Subjects were < 53 years old, had an FEV(1) of < 40% of predicted, did not have severe alpha(1)-antitrypsin deficiency, and had not undergone lung transplantation. The initial evaluation included a standardized respiratory questionnaire, spirometry, and a blood sample. A follow-up telephone interview was conducted between May and December 2002. MEASUREMENTS AND RESULTS: Subjects were young (mean age at enrollment, 47.9 years) and had severe airflow obstruction (mean baseline FEV(1), 19.4% predicted). A total of 72.7% of the subjects were women (p < 0.0001 [comparison to equal gender distribution]). The median estimated survival time was 7.0 years from the time of study enrollment, determined by the Kaplan-Meier method. The majority of deaths were due to cardiorespiratory illness. In a multivariable Cox proportional hazards model, adjusting for age, gender, and baseline FEV(1), lifetime cigarette consumption (hazard ratio [HR], 1.20 [per 10 pack-years]; 95% confidence interval [CI], 1.02 to 1.40) and recent smoking status (HR, 2.50; 95% CI, 1.03 to 6.05) were both significant predictors of mortality. CONCLUSION: In this cohort, recent smoking status predicted increased mortality independent of the effects of lifetime smoking intensity. Smoking cessation may confer a survival benefit even among patients with very severe COPD. PMID- 15539712 TI - Health-related quality of life in patients with pulmonary arterial hypertension. AB - STUDY OBJECTIVES: Patients with pulmonary arterial hypertension (PAH) often present with dyspnea and severe functional limitations, but their health-related quality of life (HRQOL) has not been studied extensively. This study describes HRQOL in a cohort of patients with PAH. DESIGN: Cross-sectional study. SETTING: A tertiary care, university hospital-based, pulmonary hypertension (PH) clinic. PARTICIPANTS: We studied HRQOL in 53 patients with PAH (mean age, 47 years; median duration of disease, 559 days). Eighty-three percent were women, 53% received epoprostenol, and 72% reported moderate-to-severe functional limitations with a New York Heart Association class 3 or 4 at enrollment. MEASUREMENTS AND RESULTS: We examined HRQOL by administering the Nottingham Health Profile, Congestive Heart Failure Questionnaire, and Hospital Anxiety and Depression Scale. We used the Visual Analog Scale and standard gamble (SG) techniques to measure preferences for current health (utilities). Compared with population norms, participants reported moderate-to-severe impairment in multiple domains of HRQOL, including physical mobility, emotional reaction, pain, energy, sleep, and social isolation. Mean SG utilities were 0.71, suggesting that, on average, participants were willing to accept a 29% risk of death in order to be cured of PH. CONCLUSIONS: PAH is a devastating condition that affects predominately young women in the prime of their life. Understanding HRQOL and preferences are important in the care and management of these patients. Compared with population norms, patients with PAH have substantial functional and emotional limitations that adversely affect their HRQOL. PMID- 15539713 TI - Normal pulmonary capillary blood volume in patients with chronic infiltrative lung disease and high pulmonary artery pressure. AB - STUDY OBJECTIVES: Pulmonary capillary blood volume (Qc), a component of diffusing capacity of the lung for carbon monoxide (Dlco), is increased in postcapillary pulmonary hypertension due to valve disease, but is decreased in primitive and thromboembolic pulmonary hypertension. This study was performed to evaluate which way pulmonary Qc is affected in patients with chronic infiltrative lung disease according to the value of systolic pulmonary artery pressure (SPAP). PATIENTS AND METHODS: Twenty-four patients who were nonsmokers and had chronic infiltrative lung disease secondary to connective tissue disease (12 patients), asbestosis (1 patient), sarcoidosis (5 patients), or of unknown origin (6 patients), and 8 control subjects underwent pulmonary function tests and Doppler echocardiography. MEASUREMENTS AND RESULTS: Total lung capacity, alveolar-arterial oxygen pressure difference, Dlco, and conductance of the alveolar-capillary membrane (Dm) did not differ between patients with low SPAP (LPAP) [ie, < 30 mm Hg] or high SPAP (HPAP). Patients with LPAP, but not HPAP, experienced significant decreases in pulmonary Qc, whatever the cause of the disease. There was a strong positive correlation between SPAP and Qc scaled by Dm to account for infiltrative disease severity (r = 0.68; p < 0.001). CONCLUSIONS: We thus conclude that pulmonary Qc is not decreased as expected in patients with chronic infiltrative lung disease and high pulmonary artery pressure. A high Qc/Dm ratio should encourage the physician to look for HPAP compatible with pulmonary hypertension, whatever the etiology of lung infiltrative disease. PMID- 15539714 TI - Is a 2-night polysomnographic study necessary in childhood sleep-related disordered breathing? AB - BACKGROUND AND OBJECTIVES: There are limited data on the night-to-night variability of childhood sleep-related disordered breathing (SDB). We aim to assess for the presence of first-night effect (FNE) and to examine whether a single-night sleep study is adequate in the assessment of childhood SDB. DESIGN: In a case-control study investigating whether obesity is a risk factor for childhood SDB, the night-to-night variability of sleep and respiratory variables were studied. PARTICIPANTS AND SETTING: Forty-six obese children from a pediatric obesity clinic and 44 age- and sex-matched normal weight control subjects from local schools. INTERVENTIONS: All subjects underwent two consecutive overnight polysomnographic studies. An obstructive apnea index (OAI) >/= 1/h was considered diagnostic of SDB. RESULTS: The mean age of the children was 11.21 years (SD 2.21). Forty-four obese children and 43 control subjects completed the 2-night study. Based on the criterion of the worst OAI over the 2 nights, 13 subjects were found to have SDB, 12 subjects were primary snorers, and 62 were normal subjects. In all subjects, the sleep efficiency improved and sleep-onset latency was reduced on the second night. While there was a rebound of rapid eye movement sleep with the associated worsening of respiratory indexes (mainly accounted for by an increase in central apneas and hypopneas) evident in normal subjects, there was a significant improvement of respiratory disturbances in the SDB group on the second night. The first-night polysomnography would have correctly identified 84.6% of cases as defined by the criteria of the worst OAI over the 2 nights. All cases missed by the first-night study had only borderline OAI. CONCLUSIONS: The phenomenon of FNE in children was well demonstrated in our study. We proposed that a single-night sleep study is adequate and more cost-effective in assessing for childhood SDB. PMID- 15539715 TI - Elevated production of tumor necrosis factor-alpha by monocytes in patients with obstructive sleep apnea syndrome. AB - STUDY OBJECTIVES: Tumor necrosis factor (TNF)-alpha is involved in the pathogenesis of atherosclerosis. In the present study, we examined TNF-alpha production by monocytes, serum levels of TNF-alpha, and the effects of nasal continuous positive airway pressure (nCPAP) in patients with obstructive sleep apnea syndrome (OSAS). DESIGN: Prospective observational study. SETTING: University hospital. SUBJECTS: Twenty-four patients with OSAS, 15 obese control subjects, and 12 healthy subjects. MEASUREMENTS AND RESULTS: After polysomnography, venous blood was collected at 5 am. Spontaneous production of TNF-alpha by monocytes for 24 h and serum levels of TNF-alpha were investigated. In addition, patients with moderate-to-severe OSAS were treated with nCPAP for 1 month, and spontaneous production of TNF-alpha by monocytes and serum levels of TNF-alpha were also measured. Spontaneous production of TNF-alpha by monocytes was significantly higher in patients with moderate-to-severe OSAS than in patients with mild OSAS (p < 0.0001), obese control subjects (p < 0.0001), or healthy subjects (p < 0.0001). Serum levels of TNF-alpha were also significantly higher in patients with moderate-to-severe OSAS than in patients with mild OSAS (p < 0.03), obese control subjects (p < 0.0005), or healthy subjects (p < 0.0001). Duration of hypoxia during total sleep time was independently associated with spontaneous production of TNF-alpha by monocytes in patients with OSAS and healthy and obese control subjects. nCPAP significantly decreased spontaneous production of TNF-alpha by monocytes (p < 0.03) and serum levels of TNF-alpha (p < 0.05) in patients with moderate-to-severe OSAS. CONCLUSIONS: Spontaneous production of TNF-alpha by monocytes and serum levels of TNF-alpha are elevated in patients with moderate-to-severe OSAS but are decreased by nCPAP. PMID- 15539716 TI - Zafirlukast treatment for acute asthma: evaluation in a randomized, double-blind, multicenter trial. AB - CONTEXT: Acute asthma causes nearly 2 million hospital emergency department (ED) visits in the United States annually, and hospitalization after an ED visit and relapse after ED discharge are common. OBJECTIVE: To evaluate the adding of therapy with zafirlukast to standardized care for patients with acute asthma in the ED and a 28-day follow-up period. DESIGN AND PATIENTS: A total of 641 patients presenting to the ED with acute asthma were randomized to receive either single-dose zafirlukast, 160 mg (Z160) [162 patients], zafirlukast, 20 mg (Z20) [158 patients]), or placebo (321 patients) as adjunct treatment to standard care in this double-blind, multicenter trial. Assessments, including spirometry and symptom scores, were obtained before each albuterol treatment and at 4 h. Patients who were discharged from the ED after 4 h continued outpatient therapy over a 28-day period and received either Z20 bid (276 patients) or placebo (270 patients) in addition to prednisone, albuterol, and their previous asthma medications. FEV(1) was measured at clinic visits on days 10 and 28. Patients recorded outpatient clinical data twice daily on a home diary card. MAIN OUTCOME MEASURES: the effect of zafirlukast on relapse after ED discharge. Other assessments were the rate of extended care (ie, ED stay for > 4 h or hospitalization), FEV(1), and symptoms. RESULTS: At the end of the outpatient period, 65 of 276 patients (23.6%) treated with zafirlukast and 78 of 270 patients (28.9%) treated with placebo relapsed (p = 0.047; absolute reduction, 5.3%; relative reduction, 18.3%). At the end of the ED period, 16 of 162 patients (9.9%) treated with Z160, 26 of 158 patients (16.5%) treated with Z20, and 48 of 321 patients (15.0%) treated with placebo required extended care (p = 0.052; absolute reduction with Z160 compared to placebo, 5.1%; relative reduction, 34%). These findings were supported by a significant improvement in FEV(1) and dyspnea in the ED with the use of Z160 therapy, and by greater improvement in FEV(1) and symptoms during the outpatient period for patients treated with Z20. CONCLUSIONS: When added to standardized care, therapy with Z20 bid reduced the risk of relapse compared with placebo over a 28-day treatment period. One dose of Z160 in the ED also reduced the rate of extended care. PMID- 15539717 TI - The prevalence of gastroesophageal reflux disease in adult asthmatics. AB - BACKGROUND: Asthma and gastroesophageal reflux disease (GERD) often coexist. However, the results of the studies investigating the prevalence of GERD among patients with asthma vary greatly. STUDY OBJECTIVE: To investigate the prevalence of GERD in adult patients with asthma. SUBJECTS AND METHODS: The basic study population consisted of 2,225 asthmatic patients who were treated in six specialist-headed hospitals during 1 year. From the common computer-based discharge register, every 14th patient was randomly selected for the study. Ninety of the 149 contacted patients (60%) agreed to participate in the study. Twenty-four-hour esophageal pH monitoring was performed on all patients. RESULTS: GERD was found in 32 of the patients (36%). Eight of these patients (25%) were free from classical reflux symptoms. Forty-seven of the 90 patients (52%) presented with typical reflux symptoms. Twenty-four of these patients (51%) were found to have abnormal acidic reflux. CONCLUSIONS: According to the current study, one third of adult patients with asthma have GERD. These patients often do not have typical reflux symptoms. However, the presence of typical reflux symptoms in an asthmatic patient does not seem to guarantee the presence of abnormal acidic reflux. PMID- 15539718 TI - Evaluation of a program aimed at increasing referrals for asthma education of patients consulting at the emergency department for acute asthma. AB - BACKGROUND: Emergency department (ED) visits for asthma may reflect poor asthma control, often due to insufficient asthma education and medical follow-up. However, few patients consulting an ED for asthma are referred for education. AIMS: To describe a model for automatic referral to educational interventions targeting patients consulting at the ED for acute asthma, to demonstrate how this model can be integrated into current care, and to increase referrals for asthma education. METHODS: The program combines a short ED-based educational intervention with the goal of motivating patients and their families to pursue an educational program with an automatic referral to an asthma education center (AEC) after agreement with ED physicians. The program was implemented in nine acute care centers with a high number of ED visits for asthma. The main study parameter was the number of patients referred to an AEC after 4 months of program implementation, as compared with 4 months before. In addition, we assessed potential barriers to successfully establishing the program. RESULTS: In the first 4 months of the program, 1,104 patients were referred to an AEC, compared with 110 for the same period the year before; 106 patients (15%) patients could not be contacted, 114 patients (16.1%) refused the intervention, 488 patients (68.9%) made appointments, and 346 patients (48.9%, or 72.8% of scheduled patients) honored their appointments. CONCLUSION: We describe a model of educational intervention for asthmatic patients consulting at the ED. We found that ED professionals can motivate patients to attend an asthma education program and that an automatic referral process is well accepted by ED staff. Such intervention can help to reduce asthma-related morbidity, but local barriers to implementation of such program should be addressed. PMID- 15539719 TI - Oximetry and indications for tracheotomy for amyotrophic lateral sclerosis. AB - STUDY OBJECTIVE: To explore the use of oximetry as a guide for using respiratory aids and tracheotomy in the treatment of patients with amyotrophic lateral sclerosis (ALS). SETTING: A retrospective review of all ALS patients presenting to a neuromuscular disease clinic since 1996. METHODS: Patients who were symptomatic for nocturnal hypoventilation were prescribed noninvasive ventilation (NIV). Patients with assisted cough peak flows of < 300 L/min were prescribed oximeters and access to mechanically assisted coughing (MAC) to prevent or reverse decreases in baseline pulse oximetric saturation (Spo(2)) levels of < 95%. The number of decreases in baseline Spo(2) that could be normalized by any combination of NIV and MAC and the duration of normalization were recorded. When the baseline was not or could not be normalized, the time to acute respiratory failure and tracheotomy or death were recorded. RESULTS: Twenty-five patients became dependent on NIV, including 13 patients who received NIV continuously for a mean (+/- SD) period of 19.7 +/- 16.9 months, without desaturation (group 1). For another 76 patients, the daytime baseline Spo(2) level decreased to < 95% 78 times. For 41 patients, the baseline level was corrected by NIV/MAC (group 2) for a mean duration of 11.1 +/- 8.7 months before desaturation reoccurred for 27 patients. Of the latter patients, 11 underwent tracheotomy, 14 died in < 2 months, and 2 had their condition again corrected by the addition of MAC therapy. For 35 patients, the desaturation was not or could not be normalized (group 3). Thirty-three of these 35 patients required tracheotomy or died within 2 months. The only significant difference between groups 1 and 2 and group 3 was significantly poorer glottic function in the patients in group 3. CONCLUSION: Tracheotomy or death is highly likely within 2 months of a decrease in baseline Spo(2) that cannot be corrected by NIV or MAC. The long-term use of NIV and MAC, and the avoidance of tracheotomy is dependent on glottic function rather than on inspiratory or expiratory muscle failure. PMID- 15539720 TI - Noninvasive ventilation by helmet or face mask in immunocompromised patients: a case-control study. AB - OBJECTIVE: To compare the efficacy of noninvasive positive pressure ventilation (NPPV) by helmet and face mask in immunocompromised patients with hypoxemic acute respiratory failure (ARF). DESIGN: Case-control study. SETTING: The general ICU of a university hospital. PATIENTS: Nineteen immunocompromised patients (hematologic malignancies [n = 8], solid-organ recipients [n = 8], AIDS [n = 3]) with hypoxemic ARF, fever, and lung infiltrates were treated with NPPV delivered by a helmet. Nineteen immunocompromised patients matched for diagnosis, age, simplified acute physiology score II, and Pao(2)/fraction of inspired oxygen (Fio(2)) receiving NPPV through a facial mask served as case-control subjects. RESULTS: The use of NPPV delivered via helmet was as effective as NPPV delivered via face mask in avoiding endotracheal intubations (intubation rate, 37% vs 47%, respectively; p = 0.37) and improving gas exchange; 14 patients (74%) in the helmet group showed a sustained improvement in Pao(2)/Fio(2) ratio (ability to increase Pao(2)/Fio(2) ratio > 200, or an increase > 100 from the baseline) in comparison with 7 patients (34%) in the mask group (p = 0.02), whose Pao(2)/Fio(2) at treatment discontinuation was higher (p = 0.02) and had fewer complications related to NPPV (ie, skin necrosis, p = 0.01). Moreover, the patients receiving ventilation via helmet required significantly less NPPV discontinuations in the first 24 h of application (p < 0.001) than patients receiving ventilation via face mask. CONCLUSIONS: The helmet may represent a valid alternative to a face mask in immunocompromised patients with lung infiltrates and hypoxemic ARF, increasing the patient's tolerance (ie, the number of hours of continuous NPPV use without interruptions) and decreasing the rate of complications directly related to the administration of NPPV. PMID- 15539721 TI - Incidence and risk factors of delayed pneumothorax after transthoracic needle biopsy of the lung. AB - STUDY OBJECTIVES: To evaluate the incidence and clinical significance of delayed pneumothorax, and to analyze the influence of multiple variables on the rate of delayed pneumothorax associated with transthoracic needle biopsy (TTNB) of the lung. STUDY DESIGN: Prospective study. SETTING: Tertiary care university hospital. STUDY SUBJECTS: Adult patients underwent TTNB from June 2001 to June 2002. MEASUREMENTS AND RESULTS: Among the 458 patients included in this study, 280 fluoroscopic-guided, 21 CT-guided, and 157 ultrasonography-guided lung biopsies were performed. A follow-up chest radiograph was obtained immediately, and 3 h, 8 h, and 24 h after the biopsy procedure. Pneumothorax that had not developed up to 3 h but developed later was defined as a delayed pneumothorax. Patients with a symptomatic or enlarged pneumothorax were treated using a pigtail catheter or chest tube. Variables such as age, gender, lesion size, location, presence of an emphysematous change, biopsy guidance methods, and biopsy devices were analyzed. Pneumothorax developed in 100 of the 458 patients (21.8%), and delayed pneumothorax developed in 15 patients (3.3%). Seventeen patients, including 3 patients with delayed pneumothorax, required a pigtail catheter or a chest tube insertion. The pigtail catheter or chest tube insertion rate in delayed pneumothorax was 20% (3 of 15 patients). Female gender and the absence of an emphysematous change correlated with an increased rate of delayed pneumothorax (p < 0.05). Lesion size, location, biopsy guidance methods, devices, and underlying diseases were not correlated with the delayed pneumothorax rate. CONCLUSIONS: The incidence of delayed pneumothorax was 3.3% of all TTNBs. Female gender and the absence of an emphysematous change were identified as risk factors for delayed pneumothorax. Delayed pneumothorax is clinically important because of its considerable incidence and the necessity for pigtail catheterization or chest tube insertion in these patients. PMID- 15539722 TI - Talc pleurodesis: basic fibroblast growth factor mediates pleural fibrosis. AB - STUDY OBJECTIVES: Patients with recurrent pleural effusions secondary to malignancy are subjected to pleurodesis if clinically indicated. Pleurodesis involves the introduction of a sclerosing agent into the pleural space. Talc is one of the most commonly used sclerosing agents in treating patients with recurrent, symptomatic malignant pleural effusions. However, the mechanisms whereby talc mediates pleural fibrosis remain unclear. We hypothesized that the intrapleural instillation of talc induces the pleural mesothelial production of basic fibroblast growth factor (bFGF), which is responsible for pleural fibrosis. METHODS: Samples of pleural fluid collected from 23 patients with malignant pleural effusions and 6 patients with congestive heart failure (control group) were included in this study. A tumor grading scale (1 to 9) was used to demonstrate the extent of the tumor. In vitro pleural mesothelial cells (PMCs) were activated with talc, and the conditioned medium was collected to evaluate bFGF levels by enzyme-linked immunosorbent assay. The bFGF-induced proliferation of fibroblasts was studied by [(3)H]thymidine incorporation. The messenger RNA expression of bFGF in talc-activated PMCs was determined by Northern analysis. RESULTS: In this study, we demonstrated that patients who have undergone successful pleurodesis following intrapleural talc insufflation have significantly higher levels of bFGF in their pleural fluid compared to those who do not respond to pleurodesis. In addition, we found a significant negative correlation between bFGF levels and tumor size. Talc-activated PMCs produce significantly higher levels of bFGF compared to control, which correlates with bFGF messenger RNA expression in PMCs stimulated with talc. The neutralization of pleural fluids and conditioned medium from talc-stimulated PMCs with bFGF antibodies significantly inhibits the bFGF-induced proliferation of pleural fibroblasts. CONCLUSIONS: An important outcome of this study was the finding that patients with extensive tumor involvement of the pleural mesothelium have a significantly lower pleural fluid bFGF response to talc compared to those who have limited involvement. Patients with limited pleural disease and higher bFGF responses go on to have successful pleurodesis, demonstrating that the presence of a mesothelium that is free of tumor enhances the possibility of success. In vitro PMCs stimulated with talc release biologically active bFGF. PMID- 15539723 TI - Intrapleural staphylococcal superantigen induces resolution of malignant pleural effusions and a survival benefit in non-small cell lung cancer. AB - BACKGROUND: Malignant pleural effusion (MPE) may occur in up to 50% of patients with non-small cell lung cancer (NSCLC). The majority of these patients have a poor performance status and a dismal prognosis, with survival duration ranging from 2 to 3 months. Since these patients are typically symptomatic from their MPE, prompt treatment is required. Patients with symptomatic MPE from NSCLC and poor performance scores (Eastern Cooperative Oncology Group [ECOG] score >/= 2, Karnofsky performance status [KPS] score < 50) are generally not offered systemic chemotherapy. Treatment is palliative and includes intrapleural catheter drainage or chemical pleurodesis with talc, doxycycline, or bleomycin. None of the latter modalities prolong survival. OBJECTIVE: Our goal was to investigate the toxicity and therapeutic effect of a new therapeutic agent, Staphylococcus aureus superantigen (SSAg), a powerful T-cell stimulant administered intrapleurally to unselected, consecutive patients with MPE from NSCLC (stage IIIb with pleural effusion) and a poor performance status. By providing direct access of the SSAg to the bronchial and mediastinal lymphatics, we predicted that intrapleural administration of SSAg would induce resolution of MPE and prolong survival in this population with advanced NSCLC and a limited prognosis. METHODS: Fourteen consecutive, unselected patients with MPE from NSCLC and a median pretreatment KPS score of 40 (range, 10 to 60) received pleural instillation of SSAg, 100 to 400 pg, once or twice weekly (mean, 3.7 +/- 1.3 treatments [+/- SD]) until the pleural effusions resolved. They were evaluated for drug toxicity, resolution, duration of MPE, and survival. RESULTS: Other than mild fever (maximum grade 2), toxicity of SSAg treatment was trivial and notably devoid of respiratory distress or hypotension. Eleven patients had a complete response (CR), and 3 patients had a partial response of their MPE. In 12 patients, the response endured for > 90 days, with a median time to recurrence of 5 months (range, 3 to 23 months). The median survival for the SSAg-treated group was 7.9 months (range, 2 to 36 months; 95% confidence interval [CI], 5.9 to 11.4 months), compared to a median survival of 2.5 months (range, 0.1 to 57 months; 95% CI, 1.3 to 3.4 months) for 18 consecutive, unselected patients with MPE from NSCLC (stage IIIb) treated with talc poudrage (p = 0.044). Survival duration of all 14 SSAg-treated cases and 13 talc-poudrage-treated patients with comparable pretreatment KPS (range, 10 to 60; median, 40 and 30, respectively), and distribution (p = 0.5) was 7.9 months (95% CI, 5.9 to 11.4 months) and 2.0 months (95% CI, 0.4 to 2.9 months), respectively (p = 0.0023). Nine of 14 patients treated with SSAg survived > 6 months, 4 patients survived > 9 months, and 3 patients survived > 350 days. One of the patients in the CR group has survived 36 months. None of the 13 talc-treated patients survived > 6 months. INTERPRETATION: In 14 unselected, consecutive patients with MPE from NSCLC and poor pretreatment performance (median KPS of 40), the intrapleural administration of SSAg was efficacious in resolving the MPE without any clinically important adverse effects. SSAg-treated patients with a median KPS of 40 (range, 10 to 60) had a median survival that exceeded that with talc poudrage, and was comparable to current systemic chemotherapy used in patients with KPS >/= 70 status. SSAg treatment is simple to perform, minimally invasive, and does not require hospital time. It may be an attractive alternative to existing palliative modalities for stage IIIb patients with MPE and poor performance who are not candidates for systemic chemotherapy. PMID- 15539724 TI - Measuring exhaled nitric oxide levels in adults: the importance of atopy and airway responsiveness. AB - BACKGROUND: Raised exhaled nitric oxide (Feno) levels have been associated with asthma. However, we have found that in children, Feno was increased in atopic children with increased airway responsiveness (AR), and this was independent of a diagnosis of asthma. STUDY OBJECTIVES: The current study was designed to test the hypothesis that in adults there is no association between Feno and asthma after controlling for atopy and AR. MEASUREMENTS: One hundred fifteen adults (77 women; mean age, 41 years) underwent an assessment that included Feno measurements, spirometry, skin-prick testing, blood eosinophil count, and inhaled histamine challenge (results are expressed as a dose-response slope [DRS]). RESULTS: When only atopic individuals were considered (n = 73), Feno was positively associated with the DRS (p = 0.003), male gender (0.02), and negatively associated with current smoking (p = 0.09). Only male gender (p = 0.03) was associated with Feno among nonatopic individuals (n = 36). In multivariate analysis, there was no association between Feno and current asthma, current wheeze, or asthma ever. CONCLUSIONS: We conclude that in adult subjects, elevated Feno measurements are associated with a phenotype characterized by atopy and increased AR regardless of the presence of asthma or asthma-like symptoms. PMID- 15539725 TI - Source of exhaled nitric oxide in primary biliary cirrhosis. AB - BACKGROUND: Exhaled nitric oxide (NO) levels may be elevated in patients with liver cirrhosis and autoimmune diseases. Primary biliary cirrhosis (PBC) is often associated with keratoconjunctivitis sicca (Sjogren syndrome [SS]), an extrahepatic autoimmune manifestation. The aim of this study was to evaluate the source of increased exhaled NO (ie, alveolar vs airway) in patients with PBC, whether associated with SS or not, and to evaluate its impact on oxygenation abnormalities. DESIGN: Observational controlled study. SETTING: University hospital. METHODS: The fractional alveolar NO concentration (FANO) and airway flux of NO (QbrNO) were measured by the multiple flows technique in 34 patients with PBC, 12 with associated SS, and were compared to 20 control subjects and 12 patients with primary SS. RESULTS: FANO was significantly higher in patients with PBC, associated with SS (mean [+/- SEM], 8.9 +/- 0.8 parts per billion [ppb]) or not (mean, 7.7 +/- 0.7 ppb) compared to healthy control subjects (mean, 4.6 +/- 0.5 ppb; p < 0.001) and to patients with primary SS (mean, 4.3 +/- 0.5 ppb; p < 0.001). FANO was significantly higher in cirrhotic patients with increased alveolar-arterial oxygen pressure difference (P[A-a]O(2)) compared to patients with normal P(A-a)O(2) values (9.8 +/- 0.8 vs 7.3 +/- 0.7, respectively; p = 0.018). When compared with control subjects and with patients with PBC not associated with SS, QbrNO was significantly increased in patients with both primary SS and SS associated with PBC. CONCLUSIONS: Increased exhaled NO levels found in PBC are from both alveolar and airway sources in patients with associated SS, but only FANO is associated with oxygenation impairment. PMID- 15539726 TI - Supplemental oxygen impairs detection of hypoventilation by pulse oximetry. AB - STUDY OBJECTIVE: This two-part study was designed to determine the effect of supplemental oxygen on the detection of hypoventilation, evidenced by a decline in oxygen saturation (Spo(2)) with pulse oximetry. DESIGN: Phase 1 was a prospective, patient-controlled, clinical trial. Phase 2 was a prospective, randomized, clinical trial. SETTING: Phase 1 took place in the operating room. Phase 2 took place in the postanesthesia care unit (PACU). PATIENTS: In phase 1, 45 patients underwent abdominal, gynecologic, urologic, and lower-extremity vascular operations. In phase 2, 288 patients were recovering from anesthesia. INTERVENTIONS: In phase 1, modeling of deliberate hypoventilation entailed decreasing by 50% the minute ventilation of patients receiving general anesthesia. Patients breathing a fraction of inspired oxygen (Fio(2)) of 0.21 (n = 25) underwent hypoventilation for up to 5 min. Patients with an Fio(2) of 0.25 (n = 10) or 0.30 (n = 10) underwent hypoventilation for 10 min. In phase 2, spontaneously breathing patients were randomized to breathe room air (n = 155) or to receive supplemental oxygen (n = 133) on arrival in the PACU. MEASUREMENTS AND RESULTS: In phase 1, end-tidal carbon dioxide and Spo(2) were measured during deliberate hypoventilation. A decrease in Spo(2) occurred only in patients who breathed room air. No decline occurred in patients with Fio(2) levels of 0.25 and 0.30. In phase 2, Spo(2) was recorded every min for up to 40 min in the PACU. Arterial desaturation (Spo(2) < 90%) was fourfold higher in patients who breathed room air than in patients who breathed supplemental oxygen (9.0% vs 2.3%, p = 0.02). CONCLUSION: Hypoventilation can be detected reliably by pulse oximetry only when patients breathe room air. In patients with spontaneous ventilation, supplemental oxygen often masked the ability to detect abnormalities in respiratory function in the PACU. Without the need for capnography and arterial blood gas analysis, pulse oximetry is a useful tool to assess ventilatory abnormalities, but only in the absence of supplemental inspired oxygen. PMID- 15539727 TI - The importance of intrapericardial drain selection in cardiac surgery. AB - INTRODUCTION: To explore the impact of intrapericardial and infracardiac drains on pericardial effusions in cardiac surgery. MATERIALS AND METHODS: Patients undergoing coronary artery bypass grafting were randomized into two groups. At the end of the intervention, an intrapericardial and infracardiac Blake drain was placed in patients in group 1 (n = 97), and an intrapericardial and infracardiac semirigid drain was placed in patients in group 2 (n = 105). In addition, a semirigid drain was placed into mediastinum in all cases. The amount of drainage was calculated at six different time points postoperatively: postoperative 0 to 2 h, postoperative 2 to 4 h, postoperative 4 to 6 h, postoperative 6 to 12 h, postoperative 12 to 24 h, and postoperative 24 to 48 h. The amount of pericardial effusion was estimated by transthoracic echocardiography after the drains were removed. RESULTS: In all measurements, the amount of drainage from intrapericardial Blake drains was higher, the total amount of drainage being equal to 330.7 +/- 29.4 mL and 193.2 +/- 19.6 mL in the Blake drain and semirigid drain groups, respectively (p = 0.000) [mean +/- SD]. When the drains were removed, the volume of pericardial effusion was 3.86 +/- 0.76 mm in Blake drain group and 7.59 +/- 1.16 mm in semirigid drain group (p = 0.000). The incidence of postoperative atrial fibrillation was 11.3% in the group with Blake drains, and 23.8% in the group with semirigid drains (p = 0.016). CONCLUSION: The more effective drainage obtained with infracardiac Blake drains compared to semirigid drains helps to reduce the amount of postoperative pericardial effusion and the risk of atrial fibrillation. PMID- 15539728 TI - Pulse pressure variation predicts fluid responsiveness following coronary artery bypass surgery. AB - STUDY OBJECTIVE: To determine whether the degree of pulse pressure variation (PPV) and systolic pressure variation (SPV) predict an increase in cardiac output (CO) in response to volume challenge in postoperative patients who have undergone coronary artery bypass grafting (CABG), and to determine whether PPV is superior to SPV in this setting. DESIGN AND SETTING: This was a prospective clinical study conducted in the cardiovascular ICU of a university hospital. PATIENTS: Twenty one patients were studied immediately after arrival in the ICU following CABG. INTERVENTION: A fluid bolus was administered to all patients. MEASUREMENTS: Hemodynamic measurements, including central venous pressure (CVP), pulmonary artery occlusion pressure (PAOP), CO (thermodilution), percentage of SPV (%SPV), and percentage of PPV (%PPV), were performed shortly after patient arrival in the ICU. Patients were given a rapid 500-mL fluid challenge, after which hemodynamic measurements were repeated. Patients whose CO increased by >/= 12% were considered to be fluid responders. The ability of different parameters to distinguish between responders and nonresponders was compared. RESULTS: In response to the volume challenge, 6 patients were responders and 15 were nonresponders. Baseline CVP and PAOP were no different between these two groups. In contrast, the %SPV and the %PPV were significantly higher in responders than in nonresponders. Receiver operating characteristic curve analysis suggested that the %PPV was the best predictor of fluid responsiveness. The ideal %PPV threshold for distinguishing responders from nonresponders was found to be 11. A PPV value of >/= 11% predicted an increase in CO with 100% sensitivity and 93% specificity. CONCLUSION: PPV and SPV can be used to predict whether or not volume expansion will increase CO in postoperative CABG patients. PPV was superior to SPV at predicting fluid responsiveness. Both of these measures were far superior to CVP and PAOP. PMID- 15539729 TI - Outcomes of COPD lung transplant recipients after lung volume reduction surgery. AB - STUDY OBJECTIVES: We sought to assess the outcomes of COPD lung transplant recipients who had previously undergone lung volume reduction surgery (LVRS), and to compare these patients to those COPD lung recipients who had not previously undergone LVRS. DESIGN: Retrospective analysis of the United Network for Organ Sharing transplant database over the period between October 25, 1999, and December 31, 2002. PATIENTS: All COPD patients who were listed and underwent transplantation during the time period were analyzed and categorized according to who did and did not have a history of LVRS. The two groups were compared for demographics, severity of illness, and various measures of outcomes after transplantation, including survival. RESULTS: There were 791 COPD patients who underwent transplantation, of whom 50 had a history of LVRS. The two groups had similar demographics and severity of disease. There was no difference in the need for reoperation, hospital length of stay, or survival between the groups. CONCLUSION: A history of LVRS does not impact on outcomes after lung transplantation and should not influence a patient's candidacy for transplantation. Similarly, a patient's potential need for lung transplantation should not impact on the decision-making process for undergoing LVRS. PMID- 15539730 TI - Colonization of dental plaques: a reservoir of respiratory pathogens for hospital acquired pneumonia in institutionalized elders. AB - STUDY OBJECTIVES: Poor dental hygiene has been linked to respiratory pathogen colonization in residents of long-term care facilities. We sought to investigate the association between dental plaque (DP) colonization and lower respiratory tract infection in hospitalized institutionalized elders using molecular genotyping. METHODS: We assessed the dental status of 49 critically ill residents of long-term care facilities requiring intensive care treatment. Plaque index scores and quantitative cultures of DPs were obtained on ICU admission. Protected BAL (PBAL) was performed on 14 patients who developed hospital-acquired pneumonia (HAP). Respiratory pathogens recovered from the PBAL fluid were compared genetically to those isolated from DPs by pulsed-field gel electrophoresis. MEASUREMENTS AND RESULTS: Twenty-eight subjects (57%) had colonization of their DPs with aerobic pathogens. Staphylococcus aureus (45%) accounted for the majority of the isolates, followed by enteric Gram-negative bacilli (42%) and Pseudomonas aeruginosa (13%). The etiology of HAP was documented in 10 patients. Of the 13 isolates recovered from PBAL fluid, nine respiratory pathogens matched genetically those recovered from the corresponding DPs of eight patients. CONCLUSIONS: These findings suggest that aerobic respiratory pathogens colonizing DPs may be an important reservoir for HAP in institutionalized elders. Future studies are needed to delineate whether daily oral hygiene in hospitalized elderly would reduce the risk of nosocomial pneumonia in this frail population. PMID- 15539731 TI - Hospital discharge to care facility: a patient-centered outcome for the evaluation of intensive care for octogenarians. AB - INTRODUCTION: Hospital survival and length of stay are commonly used for the evaluation of intensive care outcome for the young and octogenarian patients (>/= 80 years old). HYPOTHESIS: Hospital discharge to a care facility should be considered for more accurate evaluation of intensive care outcome, especially for octogenarian patients. DESIGN: An observational descriptive study. PATIENTS: A total of 6,154 consecutive hospital admissions requiring intensive care over 4 years. MEASUREMENTS: Demographics, preadmission comorbidities, severity of illness, acute hospital diagnosis categories, charges and destination after discharge, and postdischarge survival for up to 42 months. RESULTS: Octogenarians represented 15% of intensive care admissions (900 admissions). The interventions performed in the ICU, the severity of illness measured by sequential organ failure assessment (SOFA), and hospital length of stay were similar for octogenarian and younger patients. Octogenarians had higher hospital mortality (10% vs 6%, p < 0.01) and discharge to care facility (35% vs 18%, p < 0.01) than younger patients. The average hospital charge per octogenarian hospital survivor discharged to home was $128,000, compared to $100,000 for a younger hospital survivor. At follow-up, octogenarian hospital survivors who were discharged to a care facility had higher mortality than hospital survivors discharged to home (31% vs 17%, p < 0.01). On multiple logistic regression, older age, female gender, preadmission comorbidities, type of admission, SOFA score >/= 4, mechanical ventilation >/= 96 h, requirement for tracheotomy, and hospital diagnosis categories were independent factors for discharge of hospital survivors to a care facility. CONCLUSIONS: Hospital survival and length of stay did not accurately measure intensive care outcome for octogenarians. Care dependency among octogenarians who survived intensive care was prevalent and decreased their long-term survival. Care dependency and functional disability among hospital survivors should be considered for more accurate evaluation of intensive care outcome in that age group. PMID- 15539732 TI - Transthoracic echocardiography to identify or exclude cardiac cause of shock. AB - BACKGROUND: Transesophageal echocardiography (TEE) is often still considered the echocardiographic test of choice in the general ICU patient population to establish the presence or absence of cardiac cause of shock, and is often requested and performed as the initial and only echocardiographic test. This premise is based on older studies in which transthoracic echocardiography (TTE) commonly offered inadequate images in ICU patients. STUDY OBJECTIVES: We hypothesized that current TTE imaging alone is adequate to identify or exclude cardiac cause of shock in the great majority of cases. METHODS: One hundred consecutive shock cases in which an echocardiogram was requested were prospectively analyzed by two blinded echocardiographers for image adequacy, and the absence or presence of cardiac cause of shock (defined as one or more of the following: severe left ventricular (LV) or right ventricular systolic dysfunction, tamponade, severe left-sided valve disease, or a postinfarction mechanical complication), and compared to a clinical standard of presence/absence of cardiac cause of shock as determined by autopsy, surgery, or objective testing. Shock was defined as systolic BP < 100 mm Hg or fall in BP >/= 25%, and inotrope use or evidence of low output or venous congestion. Cardiac output was determined by the LV outflow tract (LVOT) Doppler method. RESULTS: Sixty-three percent of cases had a cardiac cause of shock. TTE image quality was adequate in 99% cases. Among the 99% of cases in which the imaging was adequate, the sensitivity of TTE for cardiac cause of shock was 100%, the specificity was 95%, the positive predictive value was 97%, and the negative predictive value was 100%. There were relative contraindications to TEE in 15% of cases. Stroke volume index (15 +/- 6 mL/m(2) vs 31 +/- 7 mL/m(2) [mean +/- 1 SD]; p < 0.001) and cardiac index (1.6 +/- 0.5 mL/min/m(2) vs 2.9 +/- 0.9 mL/min/m(2); p < 0.001) were significantly less in the group with a cardiac cause of shock than in the group with a noncardiac cause of shock. CONCLUSIONS: In the general critical care population, current TTE imaging identifies the great majority of cardiac causes of shock. TTE should be considered not only the initial, but also the principal echocardiographic test in the critical care environment. PMID- 15539733 TI - Predictors of long-term mortality in patients with cirrhosis of the liver admitted to a medical ICU. AB - CONTEXT: The long-term survival of patients with cirrhosis of the liver admitted to the ICU has not been described. OBJECTIVE: The main objectives were to determine the 1-year and 5-year mortality rates of a cohort of patients admitted to a medical ICU (MICU), and to identify the risk factors that may predict long term outcomes. DESIGN: This is a cohort study. We used a model-building (MB) and model validation (MV) procedure that has previously been described to determine the risk factors for overall mortality. SETTINGS: An MICU in a major referral medical center. PATIENTS: Four hundred twenty consecutive patients admitted to the ICU from January 1, 1993, through October 31, 1998, met the inclusion criteria of diagnosis of liver failure, cirrhosis, chronic liver disease, variceal bleeding, hepatic encephalopathy, or hepatorenal syndrome. Patients with acute liver failure who had undergone liver transplantation, or candidates for orthotopic liver transplantation were excluded. INTERVENTION: None. RESULTS: The 1-year mortality rate was 69%, and the 5-year mortality rate was 77%. The median survival time was 1 month. The independent predictors of mortality in patients in the MB group who retained their significance in the MV group were as follows: an acute physiology, age, and chronic health evaluation (APACHE) III score of >/= 90 (hazard ratio [HR], 2.2; 95% confidence interval [CI], 1.6 to 2.8; p < 0.0001), the use of pressors (HR, 2.5; 95% CI, 1.9 to 3.2; p < 0.0001), and jaundice (HR, 1.7; 95% CI, 1.4 to 2.2; p < 0.0001). Patients with all three risk factors (ie, APACHE III score >/= 90, use of pressors, and jaundice) had a 92% 1-month mortality rate compared to 11.2% for patients with no risk factors. CONCLUSIONS: Patients admitted to an MICU with underlying cirrhosis who are not eligible for liver transplantation have a poor long-term prognosis, even if they survive the ICU admission, particularly as the number of risk factors increases. PMID- 15539734 TI - Outcome and prognostic factors of hematopoietic stem cell transplantation recipients admitted to a medical ICU. AB - OBJECTIVE: To assess the outcome of adult hematopoietic stem cell transplantation (HSCT) recipients who were admitted to a medical ICU (MICU), and to identify the measurable predictors of their MICU outcome. DESIGN: Retrospective chart review study. SETTING: MICU in a tertiary care, university-affiliated medical center with a comprehensive cancer program. PATIENTS: Consecutive adult HSCT recipients admitted to the MICU between January 1998 and June 2001. MEASUREMENTS AND MAIN RESULTS: Eighty-five patients were admitted to the MICU, representing 11.4% of patients who had undergone HSCT during the study period. The mean (+/- SD) age at MICU admission was 46.6 +/- 11.4 years (women, 67%; men, 33%). Forty-five patients (53%) underwent allogeneic HSCT, and 40 patients (47%) underwent autologous HSCT. Fifty-one patients (60%) required mechanical ventilation (MV). Fifty-two patients (61%) survived their MICU stay, and 35 patients (41%) were discharged alive from the hospital. The long-term survival rate (ie, > 6 months) in this cohort was 28%. Nineteen mechanically ventilated patients (37%) survived their MICU stay, and 33 patients (97%) survived who did not require MV (p < 0.01). The independent predictors of poor outcome during the MICU stay were elevated serum lactate level on admission to the MICU, the need for MV, and the presence of more than two organ systems that failed. CONCLUSIONS: The study showed short-term and long-term survival rates among adult HSCT recipients who had been admitted to MICU that were higher than those previously reported. While there were no absolute predictors of mortality, patients with higher MICU admission serum lactate levels, those requiring MV, or those developing more than two organ system failures had poor MICU outcomes. PMID- 15539735 TI - The effect of an education program on the incidence of central venous catheter associated bloodstream infection in a medical ICU. AB - OBJECTIVE: To determine whether an education initiative could decrease the rate of catheter-associated bloodstream infection. DESIGN: Preintervention and postintervention observational study. SETTING: The 19-bed medical ICU in a 1,400 bed university-affiliated urban teaching hospital. PATIENTS: Between January 2000 and December 2003, all patients admitted to the medical ICU were surveyed prospectively for the development of catheter-associated bloodstream infection. INTERVENTION: A mandatory education program directed toward ICU nurses and physicians was developed by a multidisciplinary task force to highlight correct practices for the prevention of catheter-associated bloodstream infection. The program consisted of a 10-page self-study module on risk factors and practice modifications involved in catheter-related bloodstream infections and in-services at scheduled staff meetings. Each participant was required to complete a pretest before reviewing the study module and an identical test after completion of the study module. Fact sheets and posters reinforcing the information in the study module were also posted throughout the ICU. MEASUREMENTS AND MAIN RESULTS: Seventy-four episodes of catheter-associated bloodstream infection occurred in 7,879 catheter-days (9.4 per 1,000 catheter-days) in the 24 months before the introduction of the education program. Following implementation of the intervention, the rate of catheter-associated bloodstream infection decreased to 41 episodes in 7,455 catheter days (5.5 per 1,000 catheter-days) [p = 0.019]. The estimated cost savings secondary to the decreased rate of catheter-associated bloodstream infection for the 24 months following introduction of the education program was between $103,600 and $1,573,000. CONCLUSIONS: An intervention focused on the education of health-care providers on the prevention of catheter associated bloodstream infections may lead to a dramatic decrease in the incidence of primary bloodstream infections. Education programs may lead to a substantial decrease in medical-care costs and patient morbidity attributed to central venous catheterization when implemented as part of mandatory training. PMID- 15539736 TI - Comparison of breath-enhanced to breath-actuated nebulizers for rate, consistency, and efficiency. AB - OBJECTIVES: To evaluate differences between three new-generation nebulizers-Pari LC Star (Pari Respiratory Equipment; Mississauga, ON, Canada), AeroEclipse (Trudell Medical International, London, ON, Canada), and Halolite (Medic-Aid Limited, West Sussex, UK)-in terms of rate and amount of expected deposition as well as the consistency of the doses delivered. METHODS: The in vitro performance characteristics were determined and then coupled to the respiratory pattern of seven patients with cystic fibrosis (age range, 4 to 18 years) in order to calculate expected deposition. The Pari LC Star and AeroEclipse were characterized while being driven by the Pari ProNeb Ultra compressor (Pari Respiratory Equipment) for home use, and by a 50-psi medical air hospital source. The Halolite has its own self-contained compressor. Algorithms for the rate of output for the inspiratory flow were developed for each device. Patient flow patterns were divided into 5-ms epochs, and the expected deposition for each epoch was calculated from the algorithms. Summed over a breath, this allowed the calculation of the estimated deposition for each patient's particular pattern of breathing. RESULTS: The rate of deposition was highest for the Pari LC Star and lowest for the Halolite. Rate of deposition was independent of respiratory pattern for the Pari LC Star and AeroEclipse, but proportional to respiratory rate for the Halolite. The differences between the Pari LC Star and AeroEclipse were less when driven by the 50-psi source. The AeroEclipse had the least amount of drug wastage. As designed, the Halolite delivered a predetermined amount of drug very accurately, whereas expected deposition when run to dryness of the other two devices had significant variations. CONCLUSIONS: To minimize treatment time, the Pari LC Star would be best. To minimize drug wastage, the AeroEclipse would be best. To accurately deliver a specific drug dose, the Halolite would be best. PMID- 15539737 TI - In vitro exposure of bacteria to antimicrobial impregnated-central venous catheters does not directly lead to the emergence of antimicrobial resistance. AB - OBJECTIVE: Use of central venous catheters (CVCs) impregnated with minocycline and rifampin reduces the density of bacterial growth on catheters and decreases the incidence of catheter-related bloodstream infections. Questions have been raised over the possibility that the use of these catheters will lead to the emergence of antibiotic-resistant organisms. In this study, we sought to determine if in vitro exposure of four test organisms to catheter segments impregnated with minocycline and rifampin would lead to the development of antibiotic resistance. METHODS: Catheter segments (1.0 cm) were placed on the surface of agar plates previously inoculated with bacterial suspensions, such that a subconfluent lawn of colony growth would be apparent after 24 h incubation at 35 degrees C in air. Test organisms included American Type Culture Collection strains of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, and Pseudomonas aeruginosa. Zones of inhibition of colony growth surrounding catheters were measured at 24-h intervals up to 7 days (two catheter segments per test). Colonies on agar surfaces located at varying distances from catheter segments were examined for minocycline and rifampin resistance following various periods of exposure (six catheter segments per test). In addition, selected colonies were subsequently exposed to minocycline and rifampin in broth and examined for selection of minocycline and rifampin resistance (> 28 colonies per selection test). RESULTS: Inhibitory zones of 14 to 47 mm were observed with S aureus, S epidermidis, E faecalis, and E coli. Growth of P aeruginosa was not inhibited by CVC segments. Testing of colonies of the first four organisms at various distances from CVC segments after varying periods of exposure revealed only a single instance of the emergence of resistance (eg, S aureus vs rifampin). Recovery of resistant clones was enhanced with minocycline and rifampin broth selection; however, a direct link between CVC exposure and the emergence of resistance was not established. CONCLUSIONS: Our in vitro data suggest that the exposure of Gram-positive cocci to either rifampin or minocycline can lead to the development of resistance. However, exposure of bacteria to these antibiotics in combination does not directly lead to resistance. Clinical investigations will be required to determine the true risk and implications of the development of resistance. PMID- 15539738 TI - Intrapleural injection of transforming growth factor-beta antibody inhibits pleural fibrosis in empyema. AB - STUDY OBJECTIVES: Transforming growth factor (TGF)-beta is a cytokine that has been demonstrated to be an important modulator of inflammation and angiogenesis, as well as a potent stimulator of pleural fluid production and fibrosis. We previously demonstrated that rising levels of pleural fluid TGF-beta(1) correlate with pleural fibrosis in experimental empyema in rabbits. In this study, our hypothesis is that neutralization of TGF-beta with an intrapleural injection of a monoclonal antibody to TGF-beta will decrease pleural fibrosis in empyema. DESIGN: Prospective, randomized, blinded study. SETTING: Animal research laboratory. SUBJECTS: Nineteen rabbits. INTERVENTIONS: An empyema was induced in 19 rabbits by intrapleural injection of Pasteurella multocida. A panspecific monoclonal antibody to TGF-beta was injected into the pleural space on 2 subsequent concurrent days in nine rabbits. Ten rabbits received intrapleural injections of bacteria alone and served as controls. All animals were then killed on day 6. Immunohistochemistry, using the antibody to TGF-beta, was performed on pleural tissue specimens from the control rabbits. MEASUREMENTS AND RESULTS: Immunohistochemistry revealed localization of TGF-beta to macrophages in the exudative material and the visceral pleura. After injection of the antibody to TGF-beta, the amount of purulent, exudative material in the pleural space of the nine experimental animals was markedly decreased at autopsy on day 6, relative to control animals. All markers of empyema and pleural fibrosis were also significantly decreased in the rabbits receiving intrapleural anti-TGF-beta. CONCLUSIONS: TGF-beta localizes to macrophages in experimental empyema. Early intrapleural injection of an antibody to TGF-beta inhibits empyema formation and significantly decreases pleural fibrosis in experimental empyema. PMID- 15539739 TI - Oral purified bacterial extracts in chronic bronchitis and COPD: systematic review. AB - BACKGROUND: Oral lyophilized extracts of bacteria species have been used since the early 1970s to improve symptoms and to prevent exacerbations in COPD patients. The value of these treatments, which are thought to be immunomodulating, is poorly understood. Our aim was to quantify the efficacy of oral bacteria extracts in patients with chronic bronchitis and COPD. DESIGN: Systematic review of randomized trials. DATA SOURCES: Electronic databases, bibliographies, and contact with authors and manufacturers. REVIEW METHODS: Randomized comparisons of oral purified bacterial (active) extracts with placebo or no treatment (control) were selected. Meta-analyses were performed using fixed and random-effects models, and the results were expressed as relative risk (RR), odds ratio (OR), number needed to treat for one to benefit (NNTB), or number needed to treat for one to be harmed (NNTH), with 95% confidence interval (CI). RESULTS: Thirteen trials (1,971 patients), most of which were of low quality, tested OM-85BV (Broncho-Vaxom; OM Pharma; Geneva, Switzerland), LW-50020 (Luivac; ALTANA Pharma; Bad Homburg, Germany), or SL-04. Two trials (731 patients) had appropriate methodologies and reported on exacerbations. The RR in favor of the oral bacterial extract (active) was 0.83 (95% CI, 0.55 to 1.25), and the NNTB was 15.4 (95% CI, 5.5 to infinity; NNTH, 27.5). Five trials (591 patients) reported on observer-assessed improvement of symptoms RR in favor of active extracts was 0.57 (95% CI, 0.49 to 0.66), and the NNTB was 4 (95% CI, 2.8 to 5.4). Two trials (n = 344), reported on patient-assessed improvement (RR, 0.44; 95% CI, 0.31 to 0.61) [NNTB, 4; 95% CI, 3.0 to 5.9]. In two trials (163 patients), the average duration of an exacerbation was shorter with the active extracts (weighted mean difference, -2.7 days; 95% CI, -3.5 to -1.8). Itching or cutaneous eruptions was reported in 3.3% of patients (four trials; 802 patients) who received active extracts compared with 1.0% of control subjects (OR, 2.94 95% CI, 1.12 to 7.69) [NNTH, 50; 95% CI, 14 to 161]. Urologic problems (two trials; 671 patients) were reported in 8% of patients who received active extracts compared with 3.0% of control subjects (OR, 2.62; 95% CI, 1.35 to 5.11) [NNTH, 22; 95% CI, 10 to 61]. CONCLUSIONS: Oral purified bacterial extracts improve symptoms in patients with chronic bronchitis and COPD. There is not enough evidence to suggest that they prevent exacerbations. Cutaneous and urologic adverse effects are common. PMID- 15539740 TI - Seeking a home for a PET, part 3: Emerging applications of positron emission tomography imaging in the management of patients with lung cancer. AB - Positron emission tomography (PET) imaging is an important tool to refine the diagnosis and staging approach in patients with a possible lung cancer. In addition, other applications of PET imaging are being explored. Data consistently show that the intensity of uptake on a PET scan correlates with the biological aggressiveness of a tumor. PET imaging for restaging after induction therapy does not appear to be accurate enough to guide management. The results of PET imaging late after completion of treatment are highly predictive of future survival, and changes in PET images after only one cycle of chemotherapy are predictive of how a patient will respond to that planned treatment. PET imaging may allow radiotherapy treatment fields to be planned with greater accuracy, although data on how this affects patient outcomes are not yet available. Further technologic improvements in PET scanners are likely to bring further benefits to the management of patients with lung cancer in the future. PMID- 15539741 TI - Correlation between rapid strip test and the quality of sputum. AB - STUDY OBJECTIVES: To evaluate the use of a rapid strip test for the quick evaluation of sputum quality. DESIGN: Prospective, double-blind study. Sputum and saliva samples were collected. Sputum quality was assessed by the presence of polymorphonuclear neutrophils (PMNs) and squamous epithelial cells (SECs) per low power (microscopic) field (LPF) [x10 objective]. Sputum was defined as follows: (1) informative (ie, > 25 PMNs and < 10 SECs per LPF); (2) semi-informative (ie, > 25 PMNs or < 10 SECs per LPF, but not both); or (3) uninformative (ie, < 25 PMNs and > 10 SECs per LPF). The first two levels were considered to be "sputum" and the third one was considered to be "nonsputum." The quality of the sputum was compared to results obtained using a rapid strip test (Combur-Test; Roche Diagnostics; Basel, Switzerland) for specific gravity (SG), pH, leukocyte esterase (LE) activity, and levels of nitrites, protein, glucose, and erythrocytes. A Kruskal-Wallis test was used to compare the three levels of sputum quality and the rapid strip test. A Mann-Whitney test compared sputum and nonsputum to the rapid strip reagents. Pearson correlation and kappa tests were used to assess correlation. Receiver operating characteristic was used to calculate the best cut-point values, and the sensitivity and specificity of these values were calculated. RESULTS: Eighty-two samples were included, with 61 samples from hospitalized patients and 21 samples from healthy volunteers. The best predicator of sputum quality was the SG of the reagent. Using an SG threshold definition of > 1.01, the sensitivity was 86.8% and the specificity was 75.9%. The specificity of protein, glucose >/= +1, and LE levels were relatively low. No relationship was found between the results of the reagent strip test for pH, nitrites, and erythrocytes, and the sputum quality. CONCLUSION: Using an SG threshold definition of > 1.01, the rapid reagent strip test has been shown to be a sensitive test for the evaluation of sputum quality, which can be useful when facilities for sputum cytology are not available. PMID- 15539742 TI - Why do physicians who treat lung cancer get sued? AB - BACKGROUND: Minimal information exists on why malpractice actions are filed against physicians who treat lung cancer. OBJECTIVE: To review currently available data on lung cancer malpractice litigation to develop litigation avoidance strategies. DESIGN: A retrospective review of a publicly available database containing verdicts and settlements of malpractice cases. Data were then compared to the Physician Insurers Association of America (PIAA) Lung Cancer Study, which was published in 1992. The PIAA report is considered the best available data on malpractice and lung cancer. RESULTS: There were 89 patients in the current study and 213 patients in the PIAA study. Physicians are most often sued by patients in their 50s (mean age, 58.9 years; range, 34 to 80 years [current study]; vs 55 years; range, 17 to 75 years [PIAA study]). Primary care physicians (60% cases in the current study vs 33% cases in the PIAA study) and radiologists (20% cases in the current study vs 55% cases in the PIAA study) were named as defendants in > 75% of suits. Failure to diagnosis lung cancer was the most common reason physicians were sued (80% case in the current study vs 23.3% cases in the PIAA series). Despite the similarity in litigation profiles, the mean award to plaintiffs, in constant dollars, increased from $172,271 in the PIAA study to $632,261 in the current study. CONCLUSIONS: (1) Recommended strategies to avoid litigation depend on physician subspecialties. While primary care physicians would benefit most from setting up a chest radiograph tracking system, radiologists would benefit most from initiating a continuous quality improvement system to substantially decrease the misinterpretation rate of chest radiographs. (2) Over the past 12 years, there appears to have been a substantial increase in awards to patients with lung cancer who sue their physicians. However, this finding may be artificial because of differing study design. Further investigation on this subject is recommended. PMID- 15539743 TI - Is Dressler syndrome dead? AB - Post-acute myocardial infarction (AMI) syndrome was first described by Dressler in 1956. Its incidence has decreased in the reperfusion era, most likely because of the extensive use of thrombolysis and coronary balloon angioplasty, therapies that dramatically decreased the size of myocardial necrosis. The authors suggest that drugs that have been prescribed in previous decades as the post-AMI "standard-of-care," such as angiotensin-converting enzyme inhibitors, beta blockers, and statins, may also play an important role in the disappearance of Dressler syndrome due to their immunomodulatory effects. PMID- 15539744 TI - A 39-year-old man with anasarca. PMID- 15539745 TI - Ultrathin bronchoscopic barium marking with virtual bronchoscopic navigation for fluoroscopy-assisted thoracoscopic surgery. AB - STUDY OBJECTIVES: To facilitate marking and to reduce its complications, we performed barium marking using an ultrathin bronchoscope with virtual bronchoscopic (VB) navigation before thoracoscopic surgery for small pulmonary peripheral lesions. We then evaluated the feasibility, safety, and efficacy of this technique. DESIGN: A pilot study. SETTING: A tertiary teaching hospital. PATIENTS: The subjects were consecutive patients with small pulmonary peripheral lesions (ie, 180 kDa). Cleavage of alpha2-M by the enzyme resulted in the formation of 90-kDa fragments. The proteolytic activity of LV-Ka against dimethylcasein could be inhibited by alpha2-M, and the binding ratio of the inhibitor:enzyme complex was found to be 1:1. The Michaelis constant, Km, and catalytic rate constant, kcat, of LV-Ka on four selective chromogenic substrates were obtained from Lineweaver Burk plots. LV-Ka exhibits substrate specificities not only for the glandular kallikrein H-D-Val-Leu-Arg-pNA (S-2266) but also for the plasmin substrates S 2251 and Tos-Gly-Pro-Lys-pNA. Bovine kininogen incubated with LV-Ka generated a polypeptide that dose dependently contracted mesenteric arterial rings from spontaneously hypertensive rats (SHR) in a similar way as bradykinin (BK) does. As it happens with BK, LV-Ka generated polypeptide was inhibited by HOE-140, a bradykinin B2-receptor antagonist and by indomethacin, a cyclo-oxygenase inhibitor. These results strongly suggest that the polypeptide generated by LV-Ka by cleavage of bovine kininogen is bradykinin. In addition, our studies may help to understand the mechanism of action involved in hypotension produced by envenomation of bushmaster snake. PMID- 15539760 TI - Expression of cytosolic phospholipase A2 alpha in murine C12 cells, a variant of L929 cells, induces arachidonic acid release in response to phorbol myristate acetate and Ca2+ ionophores, but not to tumor necrosis factor-alpha. AB - Tumor necrosis factor-alpha (TNFalpha)-induced cell death is regulated through the release of arachidonic acid (AA) by group IVA cytosolic phospholipase A2 (cPLA2alpha) in the murine fibroblast cell line L929. However, the signaling pathway by which TNFalpha activates cPLA2alpha remained to be solved. We examined AA release in L929 cells, in a variant of L929 (C12 cells) lacking cPLA2alpha, and in C12 cells transfected with cPLA2alpha expression vectors. In transient and stable clones of C12 cells expressing cPLA2alpha, Ca2+ ionophore A23187 and phorbol myristate acetate (PMA) stimulated AA release within 90 min, although no response to TNFalpha was observed within 6 h. These results suggest that C12 cells may lack the components necessary for TNFalpha-induced AA release, in addition to cPLA2alpha. PMA is known to stimulate AA release via phosphorylation of Ser505 in cPLA2alpha by activating extracellular signal-regulated kinases (ERK1/2). However, PMA-induced AA release from C12 cells expressing mutant cPLA2alpha S505A (mutation of Ser505 to Ala), which is not phosphorylated by ERK1/2, was similar to that from L929 cells and C12 cells expressing wild-type cPLA2alpha. The role of Ser505 phosphorylation in AA release induced by PMA is also discussed. PMID- 15539761 TI - Oxazolone-induced colitis in BALB/C mice: a new method to evaluate the efficacy of therapeutic agents for ulcerative colitis. AB - A number of experimental models of colitis have been proposed. However, few studies have presented T helper-2 (Th-2) type colitis models that substitute for human ulcerative colitis (UC). In recent years, the murine oxazolone (OXA) induced colitis model came to be accepted as a Th-2 type model, but it has yet to be used in any pharmacological study. In the present study, we modified the OXA induced colitis model in BALB/C mice to evaluate the efficacy of treatments for UC. Colitis was induced by intrarectal administration of OXA solution (7.5 mg/mL in 40% ethanol) in a BALB/C strain that is known to favor Th-2 immune responses. A lower mortality rate was obtained in the BALB/C strain than was found in the original method. Histological examination showed that there were morphological similarities to human UC. Increased mRNA expression of interleukin-13, a Th-2 cytokine, was observed in mesenteric lymph nodes. Intrarectal administration of 5 aminosalicylic acid or sodium prednisolone phosphate resulted in a significant improvement in the colitis. These results suggest that the OXA-induced colitis model in the BALB/C strain provides a new way to evaluate the efficacy of therapeutic agents for UC. PMID- 15539762 TI - Molecular mechanisms and drug development in aquaporin water channel diseases: molecular mechanism of water channel aquaporin-2 trafficking. AB - Targeted positioning of water channel aquaporin-2 (AQP2) strictly regulates body water homeostasis. Trafficking of AQP2 to the apical membrane is critical for the reabsorption of water in renal collecting ducts. Besides the cAMP-mediated effect of vasopressin on AQP2 trafficking to the apical membrane, other signaling cascades also induce this sorting. Recently, AQP2-binding proteins that directly regulate this trafficking have been uncovered: SPA-1, a GTPase-activating protein (GAP) for Rap1, and cytoskeletal protein actin. This review summarizes recent advances related to the trafficking mechanism of AQP2 and its defect causing nephrogenic diabetes insipidus (NDI). PMID- 15539763 TI - Anti-inflammatory plant flavonoids and cellular action mechanisms. AB - Plant flavonoids show anti-inflammatory activity in vitro and in vivo. Although not fully understood, several action mechanisms are proposed to explain in vivo anti-inflammatory action. One of the important mechanisms is an inhibition of eicosanoid generating enzymes including phospholipase A2, cyclooxygenases, and lipoxygenases, thereby reducing the concentrations of prostanoids and leukotrienes. Recent studies have also shown that certain flavonoids, especially flavone derivatives, express their anti-inflammatory activity at least in part by modulation of proinflammatory gene expression such as cyclooxygenase-2, inducible nitric oxide synthase, and several pivotal cytokines. Due to these unique action mechanisms and significant in vivo activity, flavonoids are considered to be reasonable candidates for new anti-inflammatory drugs. To clearly establish the therapeutic value in inflammatory disorders, in vivo anti-inflammatory activity, and action mechanism of varieties of flavonoids need to be further elucidated. This review summarizes the effect of flavonoids on eicosanoid and nitric oxide generating enzymes and the effect on expression of proinflammatory genes. In vivo anti-inflammatory activity is also discussed. As natural modulators of proinflammatory gene expression, certain flavonoids have a potential for new anti inflammatory agents. PMID- 15539764 TI - Interleukin-1 balance in the lungs of preterm infants who develop bronchopulmonary dysplasia. AB - BACKGROUND: The local pulmonary balance between the agonist and antagonist of interleukin-1 (IL-1) may influence the development of inflammatory disease and resultant structural damage in a variety of human diseases including adult respiratory distress syndrome and asthma. OBJECTIVES: We tested the hypothesis that IL-1 cytokines are early markers for bronchopulmonary dysplasia (BPD), when measured in tracheal aspirates (TAs) obtained from premature infants being ventilated for respiratory distress syndrome during the first week of life. METHODS: Serial TAs were collected on days 1, 3, 5 and 7 from 35 preterm infants (16 BPD, 19 non-BPD) in the absence of chorioamnionitis, and were assayed for IL 1 cytokines and leukocytes. RESULTS: In spite of comparable maternal demographic and clinical characteristics, premature infants who developed BPD had higher levels of IL-1 receptor antagonist (Ra) in their airways on the first day of life. This antagonist IL-1Ra was an early and persistent marker for BPD during the first week of life. The agonist IL-1beta also increased significantly for BPD patients early, both compared to non-BPD patients, and also within the BPD group. While the early (day 1) IL-1 antagonist/agonist molar balance offered protection, by days 5 and 7, a threshold for IL-1Ra in the presence of increasing IL-1beta expression-favored pro-inflammation in the BPD group. CONCLUSIONS: We conclude that a strong and early expression of airway antagonist (IL-1Ra) proves ultimately to be sub-optimal and non-protective due to the robust expression of airway agonist (IL-1beta) seen by day 5 in premature infants who develop BPD. PMID- 15539765 TI - Hyperbilirubinemic serum is cytotoxic and induces apoptosis in murine astrocytes. AB - BACKGROUND: High levels of unconjugated bilirubin can be neurotoxic and gliotoxic. However, the effect of serum from patients with neonatal indirect hyperbilirubinemia on astrocyte viability has never been investigated. OBJECTIVES: In the present study, we searched for the possible toxic effect of hyperbilirubinemic serum on murine astrocytes. METHODS: Heat-inactivated patient serum was added to astrocyte cultures at different concentrations varying from 1 to 20%, and cultures were incubated for 24, 48, and 72 h. Sera from healthy infants without hyperbilirubinemia were used as controls. Cytotoxicity was evaluated according to the release of lactate dehydrogenase in the culture medium. Apoptotic cell death was determined by anti-single-strand DNA immunostaining. RESULTS: The results of the present study show that hyperbilirubinemic serum induces cytotoxicity and apoptotic astrocyte death in a concentration- and time-dependent manner. CONCLUSIONS: We conclude that serum from patients with neonatal indirect hyperbilirubinemia is cytotoxic to murine astrocytes. PMID- 15539767 TI - Clinical trials for evaluating diagnostic markers of infection in neonates. PMID- 15539766 TI - Serum amyloid A protein is a useful inflammatory marker during late-onset sepsis in preterm infants. AB - BACKGROUND: Few studies demonstrated that serum amyloid A (SAA), a non-specific acute-phase reactant, could be used as a reliable early marker for the diagnosis of late-onset sepsis (LOS). OBJECTIVES: To evaluate the diagnostic value and the dynamics of SAA levels during the course of LOS and to compare it to those of other inflammatory markers. METHODS: Levels of SAA, C-reactive protein (CRP) and IL-6 together with clinical variables, biochemical parameters and cultures retrieved from all preterm infants suspected of LOS were checked at the first suspicion of sepsis and after 8, 24, 48 and 72 h. Results were compared to healthy, matched infants. RESULTS: One hundred and sixteen infants were included in the study, 38 in the sepsis and 78 in the non-sepsis group. High levels of SAA were observed at sepsis onset, with a gradual decline thereafter, while CRP levels increased only at 24 h after sepsis onset. In the sepsis group, levels of SAA returned faster to baseline than CRP levels. Receiver-operating characteristic analysis values revealed that SAA at 10 mug/ml had the highest sensitivity at 0, 8 and 24 h after sepsis onset (95, 100 and 97%, respectively) and a negative predictive value (97, 100 and 98%, respectively). CONCLUSIONS: SAA is an accurate acute-phase protein during LOS in preterm infants. Quick and reliable SAA kits can make this marker a useful tool in LOS in preterm infants. PMID- 15539768 TI - Dexamethasone-induced prenatal alveolar wall thinning is associated with a decrease in EIIIA+ fibronectin isoform in the fetal rat lung. AB - BACKGROUND: Glucocorticoid hormones play an important role in architectural and biochemical lung maturation. Although much of the molecular mechanism of their action in the lung is not fully understood, glucocorticoids directly or indirectly regulate lung maturation. Indirect effects of glucocorticoids may involve the modulation of cell-cell or cell-matrix interactions. Fibronectin (FN) is the major constituent of the pulmonary extracellular matrix and exists in multiple isoforms arising from alternative RNA splicing. EIIIA is the major alternatively spliced segment, and its expression is regulated in a spatiotemporal and oncodevelopmental manner. OBJECTIVES: The present study focuses on the regulation of EIIIA-containing FN isoforms (referred to as EIIIA+ FN) by glucocorticoids in the developing lung. METHODS: Dexamethasone (DEX) or saline was injected daily into pregnant rats from day 15 of gestation (term = day 22) until 24 h before sacrifice. The expression of EIIIA+ FN and proliferating cell nuclear antigen (PCNA), a biochemical marker for cell proliferation, was investigated in the fetal rat lung. RESULTS: At day 20 of gestation (the canalicular stage), the DEX-treated lung showed a significant decrease in weight and saccular septal wall thickness, while the messenger RNA expression of the surfactant protein SP-B was increased in the DEX-treated lung, as compared with the control lung. The expression of EIIIA+ FN and PCNA around the distal airspaces was less extensive in the DEX-treated lung than in the control lung at day 20 of gestation. CONCLUSIONS: Given the finding in vitro that EIIIA+ FN regulated the cell cycle, our results suggest that the change of EIIIA+ FN expression in the DEX-treated lung affected pulmonary cell proliferation. PMID- 15539769 TI - Antioxidant status of neonates exposed in utero to tobacco smoke. AB - To investigate the influence of maternal smoke exposure on neonatal and maternal antioxidant status, 39 mothers who were active smokers, 14 mothers exposed to environmental tobacco smoke (ETS), 17 controls, and their newborns were included in a prospective, controlled study. Plasma total antioxidant capacity, measured as total radical-trapping antioxidant parameter (TRAP) and ferric reducing antioxidant power (FRAP), and concentrations of specific antioxidants were measured in cord and in maternal blood. A similar, significant increase in ceruloplasmin concentration was observed in neonates born to actively smoking mothers and in those born to ETS exposed mothers. Uric acid and TRAP concentrations were significantly increased in ETS-exposed newborns and their mothers, compared to newborns and mothers from the active smoking and no-exposure groups with a trend towards increased uric acid, TRAP and FRAP concentrations being observed in the active smokers group. Neonatal and maternal antioxidant concentrations correlated significantly, except for ceruloplasmin. Cord blood vitamin A, E and C concentrations were unaffected by smoke exposure. These results show that maternal active smoking as well as ETS exposure significantly affect neonatal and maternal antioxidant status. PMID- 15539770 TI - Richard Morton: origins of anorexia nervosa. AB - Sir William Gull coined the name 'anorexia nervosa'. Examples of self-starvation appeared in the Hellenistic era. Holy anorexics abused their bodies, rejected marriage and sought religious asylum where many perished and became saints. The condition then paled into obscurity until the 19th century. Louis-Victor Marce (1828-1864) described such a patient in 1859, but Richard Morton is generally credited with the first medical description of anorexia nervosa in 1689. Two neurologists in 1873 separately described anorexia nervosa. Ernest Charles Lasegue, a student friend of Claude Bernard, and a favourite pupil of Trousseau wrote of a refusal of food that may be indefinitely prolonged. Historical precedence is explored and citations included. PMID- 15539771 TI - Mechanism of disorientation in Alzheimer's disease. AB - In nondemented amnesics, disorientation mostly reflects a failure to select memories that pertain to ongoing reality (increased temporal context confusion; TCC), a failure strongly associated with orbitofrontal damage. In the present study, we used the same paradigm--2 runs of a continuous recognition task--in 23 patients with mild to moderate Alzheimer's disease (AD) (Clinical Dementia Rating Scale score of 1-2). We found that disorientation was frequent in this sample (52%). However, although it correlated moderately well with general measures of dementia severity, verbal episodic memory and executive functioning, there was no significant correlation with TCC. Thus, disorientation in AD appears to reflect general cognitive decline rather than a specific cognitive failure such as increased TCC. This finding is compatible with the different distribution of degeneration in AD and the orbitofrontal damage typically present in severely disoriented amnesics. PMID- 15539772 TI - Different perceived foreign accents in one patient after prerolandic hematoma. AB - Foreign accent syndrome (FAS), a rare disorder characterized by the emergence of a new accent perceived as foreign by listeners, is usually reported with left brain damage. We here report the case of a 28-year-old native Brazilian who appeared, to the examiner, to show a North American accent during recovery from Broca's aphasia. The lesion was due to a frontal hematoma. Without referring specifically to speech, we asked 10 independent observers to comment on a videotape of the patient's interview. Seven reported that the patient had a foreign accent, while 3 simply noted a 'strange' accent. The observers did not agree on the origin of the accent, 5 identifying it as Spanish, 1 as German, and 1 as south Brazilian. These findings suggest that FAS is not due to the acquisition of a specific foreign accent, but to impairment of the suprasegmental linguistic abilities (tone, accent, pauses, rhythm, and vocal stress) that make it possible to distinguish native language. PMID- 15539773 TI - Interferon Beta-1a and intravenous immunoglobulin treatment for multiple sclerosis in Iran. AB - The aim of the study was to evaluate the efficacy and safety of interferon beta 1a (Avonex) and intravenous immunoglobulin (IVIG) in clinical practice for the treatment of relapsing-remitting multiple sclerosis. Avonex is the most common disease-modifying therapy used in Iran due to its ease of administration. IVIG is also frequently used due to its alleged effectiveness and fewer side effects. Eighty patients were selected and prospectively monitored according to a predefined protocol. They were then randomized to receive either weekly intramuscular injections of Avonex or 0.4 g/kg monthly IVIG in a single blind fashion and following an attack of exacerbation which was treated with steroids. Basal relapse rate and Expanded Disability Status Scale (EDSS) were similar in both groups of patients (p > 0.4). Seventy-two patients remained in the study. The annual relapse rate consistently decreased from 0.95 +/- 0.41 to 0.60 +/- 0.67 (approximately 32%, p < 0.001) for 34 patients treated with Avonex and from 1.05 +/- 0.34 to 0.55 +/- 0.46 for 38 patients in the IVIG group (approximately 47%, p < 0.001). EDSS decreased by 0.4 units in IVIG-treated patients (p < 0.001) and remained stable (0.2 < p < 0.3) in the Avonex arm. This study confirms the relative efficacy of both treatments with better safety profile for IVIG in the studied Iranian population. However, the results are very preliminary ones, due to limited numbers of patients and only 12 months of treatment. PMID- 15539774 TI - Cervical artery dissection: a 5-year prospective study in the Belluno district. AB - BACKGROUND: Artery dissection is an unusual cause of ischemic stroke, particularly frequent among young patients. The aim of this study was to collect epidemiological data on artery dissection in a hospital-based community, set up a diagnostic protocol and discover outcome predictors. METHODS: Among patients suffering from cerebral infarction resident in our country, those with clinical and radiological features suggestive of artery dissection were selected. Risk factors, investigative techniques and treatment were evaluated. Patients were subjected to clinical examinations and were regularly tested neuradiologically. RESULTS: Out of 895 ischemic stroke patients, 10 patients with cervical artery dissection (1.1%) were found. Seven patients were treated with anticoagulants and 3 received antiplatelet agents. One posttraumatic artery dissection patient died within a few days of the stroke. None of the patients suffered from a recurrence, while serious disability occurred in 4 of them. CONCLUSIONS: Artery dissection should be suspected in any cerebral infarction patient, especially in young patients without risk factors for cerebrovascular diseases. The treatment of choice consists of anticoagulants. An early clinical diagnosis, strongly supported by radiological tests, is mandatory to start the proper treatment and achieve the best possible outcome. PMID- 15539775 TI - Central representation of somatic sensations in the parietal operculum (SII) and insula. AB - Four subjects with small restricted cerebral cortical infarcts have been examined. One had a lesion confined to the parietal operculum (SII), while in the second the SII lesion also encroached on the posterior insula; in the third subject, both banks of the sylvian fissure and the dorsal insula were involved, while in the fourth the lesion involved the upper bank of the sylvian fissure. In all cases, the postcentral gyrus (SI) was intact. Subjects 1 and 2 had mild spontaneous pain, but subjects 3 and 4 had never had spontaneous pain. In the affected areas, none could feel mechanical (skinfold pinch) pain. The 2 subjects with spontaneous pain could not discriminate sharpness (pinprick), but this was unimpaired in the third and fourth subjects. Warmth, cold, and heat pain were impaired in the 2 subjects with spontaneous pain, but not in those without; however warm-cold difference was greater in the affected regions of all subjects. The possibility must nevertheless be considered that the presence of central pain in some way alters the cortical mechanisms for the perception of thermal stimuli. Certainly, as we had earlier observed, spontaneous pain only occurs when there is interference with thermal sensation. Functional MRI (fMRI) studies following thermal stimulation in subjects 1 and 2 showed these areas, particularly SII, to be concerned with the reception of innocuous and noxious thermal stimuli, mechanical (skinfold pinch) pain and sharpness (pinprick), implying that SI is principally concerned with the reception of low-intensity mechanical stimuli, although it was activated in 1 of our fMRI-studied subjects by innocuous cooling. PMID- 15539776 TI - Does dissociation offer a useful explanation for psychopathology? AB - BACKGROUND: Dissociation is often conceptualised as an altered state of consciousness, a trance-like state in which normal barriers between conscious and unconscious memories, desires and beliefs break down and other amnestic barriers emerge. This review explores whether it is likely that there is a neurophysiology of pathological dissociative processes that will elucidate management. METHOD: A critical reading of current research, sourced through Medline and Psychinfo searches from 1990 to 2002, using subject headings: dissociative disorders, hypnosis and stress disorder (post-traumatic), as well as keywords: dissociation, hypnosis and trance. RESULTS: Current knowledge does not support the notion of dissociation as a discrete brain state or process. CONCLUSIONS: Psychiatric and neurophysiological research and theory development are better directed towards individual components that contribute to dissociative experience. PMID- 15539777 TI - Facial expression and experience of emotions in psychodynamic interviews with patients suffering from a pain disorder. Indicators of disorders in self- and relationship regulation in the involuntaryfacial expression of emotions. AB - In clinical practice, it is often difficult to establish a positive relationship with patients suffering from psychogenic pain disorder. The present study addresses this maladaptive relationship pattern that is characterized by a mixture of disappointment and rejection. Facial expressions of emotions of 21 female patients in psychodynamic interviews were analyzed and compared to a control group of 11 healthy subjects. The assumption of a maladaptive relationship pattern is supported by an increase in negative facial expressions, especially contempt, on the side of the patients and the interviewer. These negative emotions indicate a disorder of self- and relationship regulation and are negatively correlated with the level of structural personality organization (as measured by the operationalized psychodynamic diagnostic). They also correlate with the emotional experience of shame and fear and are negatively correlated with the experience of joy. The results demonstrate how maladaptive relationship patterns are implemented at the level of involuntary emotional behavior. They can help to understand and improve the emotional quality of therapeutic relationships. PMID- 15539778 TI - Interactive regulation of affect in postpartum depressed mothers and their infants: an overview. AB - Specific patterns of interaction emerging in the first months of life are related to processes regulating mutual affects in the mother-child dyad. Particularly important for the dyad are the matching and interactive repair processes. The interaction between postpartum depressed mothers and their children is characterized by a lack of responsiveness, by passivity or intrusiveness, withdrawal and avoidance, as well as a low level of positive expression of affect. Thus, an impaired capability to regulate the child's affect has been demonstrated in depressed mothers. Maternal aggression, neglect toward infants, infanticidal thoughts, as well as infanticidal behavior are mainly linked to severe postpartum depression, especially with psychotic symptoms. The findings on mother-child interaction reported in this paper are based on mothers with mild to moderate depressive disorders without psychotic symptoms. Considering the stability of interaction patterns in the course of depressive illness as well as the long-term consequences of these interactions, it seems surprising that there are still few systematic studies of depressed mothers interacting with their infants. In connection with an overview on these issues, treatment models for parent-infant psychotherapy are discussed. PMID- 15539779 TI - Dissociative symptoms in schizophrenia: a comparative analysis of patients with borderline personality disorder and healthy controls. AB - The aim of this study was to investigate the occurrence of dissociative symptoms in patients with a schizophrenic disorder. The pattern of dissociative experiences was examined in a group of patients with a diagnosis of the schizophrenia spectrum disorder (n = 26; mean age 27.9 years), and a group of patients with a diagnosis of borderline personality disorder (n = 26; mean age 24.0 years) was compared with normal controls of the general population (n = 1,056; mean age 18.7 years). The degree of self-reported dissociative symptoms was measured using the German version of the Dissociative Experiences Scales. The dissociation scores were significantly higher among patients with a diagnosis of borderline personality disorder compared to the group of schizophrenic patients and to the control group. There was no difference in the degree of reported dissociative experiences between the group of schizophrenic patients and the normal volunteers. An analysis of the subdimensions (dissociative amnesia, absorption/imaginative involvement, depersonalization/derealization) of the scale revealed the same strong distinctions between the investigated groups. There was no evidence that dissociative symptoms reflect a specific vulnerability in young schizophrenic patients. PMID- 15539780 TI - Distribution of symptoms of conduct disorder and antisocial personality disorder in Turkey. AB - BACKGROUND: A variety of conduct disorder (CD) symptoms are seen together as a symptom cluster. Among CD symptoms there are serious and stubborn antisocial behaviors: lying, swindling, running away from home/school, destructiveness, arson, kidnapping women, sexual abuse, and armed robbery. The objective of this study was to discover the relationship between CD and antisocial personality disorder (APD) and the distribution of the symptoms of these disorders. SAMPLING AND METHODS: The research sample consisted of 994 individuals between the ages of 18 and 65 years in Sivas province. Subjects were given the Diagnostic Interview Schedule-III-R (DIS-III-R) APD subscale. RESULTS: The lifetime prevalence for CD and APD was found to be 21.03 and 3.02%, respectively. In both disorders the percentage of men was higher than for women. The comorbidity of both disorders was found to be 14.35%. The most frequently seen CD symptom in normal society is 'lying', but in those with CD 'initiates a fight several times' is the most frequently seen symptom. In those with APD the most frequently seen symptom is 'truant from school several times'. CONCLUSIONS: Identifying risk factors in children could be an important step in preventing the progression of CD and APD. PMID- 15539781 TI - Psychiatric conditions in an evolutionary context. AB - Psychiatric conditions and the institutions and practices that modern society has evolved to handle them originated during the nineteenth century in Anglo European societies. They are products of a historically contingent and culture specific formulation of a class of social problems of behavior that came to the fore in relation to intellectual and political economic changes of those societies. However, such problems have a long ancestry. They are intrinsic to human species and the social and cultural systems that its members have evolved since their emergence. This article reviews intellectual quandaries raised by evolutionary study of psychiatric conditions, those of crossing the human/animal divide and crossing historically contingent cultures; and of framing history of psychiatry in terms of social and cultural evolution. The biological architecture underlying psychiatric conditions and the breakthroughs that indigenous psychiatry of different types of societies underwent in formulating signs and symptoms are discussed. PMID- 15539782 TI - LMNA mutations in cardiac transplant recipients. AB - Lamin A and C are components of the nuclear envelope, located at the nucleoplasmatic surface of the inner nuclear membrane within cells. Recently, mutations within LMNA encoding lamin A/C have been associated with various disease entities including cardiomyopathy. We screened heart transplant recipients suffering from dilated cardiomyopathy (DCM) with a positive family history of LMNA mutations. Four index patients and one relative belonging to four unrelated families carrying LMNA mutations were identified. The mutations p.Q355X and p.S22L have not been reported before, whereas p.R190W has already been reported in other studied DCM cohorts. In the patients of the present study, the mean age at manifestation of heart disease was 37.6 years (range 30-45 years), with progression to end-stage heart failure requiring transplantation at a mean age of 45.8 years (range 35-54 years). Three patients presented initially with atrial fibrillation. These data confirm the involvement of LMNA mutations in patients with DCM and extend the mutational spectrum of LMNA. The p.R190W mutation has been reported in different populations and may therefore be useful for analyzing the impact of a specific LMNA mutation on the phenotype of muscle disease. PMID- 15539784 TI - Left ventricular hypertrabeculation/noncompaction and stroke or embolism. AB - Left ventricular hypertrabeculation/noncompaction (LVHT) is reported to be associated with embolism. This retrospective study assessed the number of strokes/embolisms in 62 patients with LVHT and in 62 control patients matched with regard to age, sex and left ventricular systolic function. The incidence of strokes or embolism was 10% in patients with LVHT and 15% in controls. LVHT by itself does not seem to be a risk factor for stroke or embolism and thus is not an indication for oral anticoagulation. However, if LVHT is associated with systolic dysfunction, oral anticoagulation should be considered. PMID- 15539783 TI - Relationship of serum cardiac markers following successful percutaneous coronary intervention and subsequent exercise capacity in patients with chronic stable angina: a pilot study. AB - We performed a pilot study to assess the significance of modest, asymptomatic, elevated serum cardiac markers--troponin T (TnT) and creatinine kinase-MB (CK-MB) -18-24 h following successful elective percutaneous coronary intervention (PCI) and to explore their relationship with changes in aerobic exercise capacity, as measured by peak oxygen consumption (VO2), 6 weeks following the procedure. Twenty-three patients with single-vessel disease and chronic angina performed an incremental cardiopulmonary exercise test before and 6 weeks after successful uncomplicated PCI. A venous blood sample was taken at rest before PCI and 18-24 h after the procedure. Successful PCI resulted in a trend towards an increased peak VO2 [21.62 +/- 0.64 (pre) vs. 23.03 +/- 0.75 ml/ kg/min (post), p = 0.07; mean +/ SEM]. There was a significant increase [median (IQR)] in TnT, from 0.00 (0.00) microg/l at baseline increasing to 0.02 (0.03) microg/l at 18-24 h, p = 0.002. CK MB levels showed no significant difference. In the group of 15/23 (65%) patients with an elevation in serum TnT (>/=0.01 microg/l), 18-24 h after successful PCI, there was no significant increase in peak VO2 [23.31 +/- 0.96 (pre) vs. 23.89 +/- 1.09 ml/kg/min (post), p = 0.57]. In 8 (35%) patients with no rise in TnT at 18 24 h, a significant increase in peak VO2 was observed following successful PCI [23.10 +/- 0.91 (pre) vs. 25.09 +/- 0.75 ml/kg/min (post), p = 0.02]. Although 7 of these 8 patients increased their peak VO2, the absence of a TnT rise at 18-24 h was not significantly associated with an increase in peak VO2 following successful PCI (p = 0.18). To confirm these interesting initial results and investigate the relationship of serum cardiac markers following successful PCI and subsequent exercise capacity, further studies are required. PMID- 15539785 TI - Normalization of negative T waves in the chronic stage of Q wave anterior myocardial infarction as a predictor of myocardial viability. AB - We investigated whether spontaneous normalization of negative T waves (TWN) on infarct-related ECG leads (IRLs) in the chronic phase of Q wave anterior myocardial infarction (MI) could be a predictor of residual viability in infarct areas. We prospectively studied 35 patients (age 60 +/- 8.6 years) in the chronic phase of Q wave anterior MI. Spontaneous TWN (group A, n = 23) were defined as negative T waves that became upright (> or =0.15 mV) in > or =2 IRLs. The presence of negative T waves (group B, n = 12) was defined as symmetric or biphasic negative T wave of > or =0.15 mV. All patients underwent same-day rest 201Tl-stress (99m)Tc sestamibi dual-isotope myocardial perfusion SPECT and 24 hour 201Tl reinjection imaging for ischemia and viability analysis. On scintigraphic examination, ischemic or viable myocardial segments were found in 18 patients (78%) with TWN and 4 patients (33%) of group B (p = 0.013). The use of TWN as a parameter had a marked influence on the sensitivity (82%), specificity (62%), positive (78%) and negative (67%) predictive values and accuracy (74%) of the diagnosis of viable myocardium. If we add the criterion of positive T waves in aVR with negative T waves to our criteria, we found that sensitivity (90%), positive (80%) and negative (80%) predictive values and accuracy (80%) increased. The results of our study suggest that analysis of TWN on IRLs is an accurate marker of residual viability and/or persistent peri infarct ischemia in patients in the chronic stage of Q wave anterior MI, and therefore optimizes the diagnostic and therapeutic strategies after MI. PMID- 15539786 TI - Is there a seasonal variation in hospital admissions for acute-onset atrial fibrillation? PMID- 15539787 TI - One year of clinical experience in postdilution hemofiltration with online reinfusion of regenerated ultrafiltrate. AB - BACKGROUND/AIMS: Hemofiltrate reinfusion (HFR) is characterized by the use of regenerated ultrafiltrate as replacement fluid. We set up a new technique, postdilution HFR (PD-HFR), aiming at increasing purification efficiency, treatment tolerance and at reducing inflammatory states. METHODS: We performed PD HFR in 6 uremic patients during 1 year. Dialysis efficacy, dialyzer blood loss and the behavior of cytokines were evaluated. RESULTS: No pyrogenic reactions or other adverse events were recorded. Treatment tolerance was excellent. We observed high urea extraction rates and optimal Kt/V values, high beta2 microglobulin (beta2m) extraction rates and a decrease in dialyzer blood loss; also IL-6 and TNF-alpha decreased significantly. CONCLUSIONS: Inversion of the standard HFR configuration has allowed us to improve the removal of both urea and beta2m, and to decrease dialyzer blood loss, with an optimal tolerance. Moreover, the decrease in cytokine levels might attenuate the uremic microinflammatory state. PMID- 15539788 TI - Vitamin E-coated dialyzer membranes downregulate expression of monocyte adhesion and co-stimulatory molecules. AB - BACKGROUND: In patients on chronic hemodialysis leukocyte activation has been related to the impaired function of the immune system. In this study we investigated if the vitamin E-coated dialyzer membrane could reduce monocyte activation thereby improving cellular immunity. METHODS: This hypothesis was tested in a prospective crossover trial in which 14 stable hemodialysis patients were switched from the baseline hemophane dialyzer to a vitamin E-coated and thereafter a polysulphone dialyzer membrane or vice versa. RESULTS: Monocyte MHC class I, CD54 and ICAM-1 expression was significantly downregulated when a vitamin E-coated or polysulphone dialyzer was used. The use of a vitamin E membrane specifically decreased monocyte CD40 and CD86 expression. Lectin induced T cell proliferation increased with the use of the vitamin E-coated membrane as compared to polysulphone and hemophane dialyzers. CONCLUSION: Vitamin E-coated dialyzers induced a less-activated phenotype of monocytes and may improve cellular immunity. PMID- 15539789 TI - Effect of capsaicin on bile secretion in the rat. AB - Capsaicin is a popular food ingredient. This study aimed to determine if capsaicin can affect bile flow and bile protein secretion, and the extent to which capsaicin can reach bile in the rat. The effect of capsaicin was studied in anesthetized rats equipped with cannulas inserted into the common bile duct. Capsaicin was administered by intragastric, intraduodenal (4-400 microg/ml; 10 1,000 microg/kg), intravenous (10 microg/kg) routes or applied on the serosal surface of the duodenum at 4 microg/ml. The administration of capsaicin decreased bile flow in comparison to the corresponding basal values, the maximum effect being reached at a concentration of 400 microg/ml of intragastric capsaicin (30.2%; p < 0.01), and 40 microg/ml (30.8%; p < 0.01) of intraduodenal capsaicin, 75 min after drug administration. Meanwhile, a decrease of 24.7 and 40% was observed 60 min after serosal and intravenous capsaicin administration, respectively. Biliary protein secretion was also reduced following capsaicin administration. High-performance liquid chromatography (HPLC) analysis showed that capsaicin was readily excreted into bile, peak levels were reached 75 or 60 min following its intragastric or intraduodenal administration, respectively (range 100-248 and 144-698.6 ng/ml after intragastric or intraduodenal capsaicin at 4-400 microg/ml, respectively). Capsaicin concentrations of 86 and 75 ng/ml could be detected in bile 15 min after intravenous administration or serosal application of the agent, respectively. Results suggest that capsaicin is readily absorbed after its intragastric or intraduodenal administration and goes through hepatobiliary excretion. Results also indicate that administration of capsaicin reduces bile flow and biliary proteins in the rat. PMID- 15539790 TI - Antihypertensive effect of 5-HT1A agonist buspirone and 5-HT2B antagonists in experimentally induced hypertension in rats. AB - We investigated the antihypertensive effect of 5-HT1A agonist (buspirone) and 5 HT2B antagonists (SB204741 and SB200646) in deoxycorticosterone acetate (DOCA) salt-induced hypertensive rats. Experiments were divided into two sets: in the first set, sham-operated control and DOCA-treated hypertensive rats received buspirone (1 mg/kg/day p.o. for 4 weeks) and in the second set, in vivo and in vitro studies were carried out. In the case of in vivo studies, sham-operated control and DOCA-treated hypertensive rats received SB204741 or SB200646 (1 mg/kg/week i.v. for 4 weeks). Blood pressure was measured weekly by tail-cuff method. After completion of the treatment schedule, blood pressure and vascular reactivity to various agonists like 5-HT, noradrenaline and adrenaline were recorded. Chronic administration of buspirone, SB204741 and SB200646 produced a significant reduction in blood pressure and vascular reactivity to agonists in DOCA-salt hypertensive rats, implying an antihypertensive effect. However, chronic administration of the same drugs in sham control rats did not alter blood pressure and vascular reactivity to various agonists. For in vitro studies a similar treatment schedule was followed as in vivo studies and a cumulative concentration response curve of 5-HT was recorded on isolated thoracic aorta. Treatment with 5-HT2B antagonists shifted the concentration response curve of 5 HT to the right on isolated aorta. We conclude that 5-HT1A agonist and 5-HT2B antagonists possess an antihypertensive effect. PMID- 15539791 TI - Effect of angiotensin II on glomerular structure in streptozotocin-induced diabetic rats. AB - BACKGROUND/AIMS: The streptozotocin (STZ)-induced diabetic rat is a widely used animal model of human diabetic nephropathy. In this model, diabetic nephropathy progresses without significant elevation in blood pressure. Therefore, studies have examined the effect of hypertension in STZ spontaneously hypertensive rats (SHR). This study investigated angiotensin II (Ang II)-induced hypertension in diabetic nephropathy in the STZ-diabetic rat independent of deleterious genetic effects in SHR. METHODS: Animals were divided as follows: nondiabetic controls (ND; n = 18); diabetic (STZ: 65 mg/kg; n = 16); Ang II-induced hypertensive ND (Ang II: 120 ng/kg/min; n = 9), and hypertensive diabetic rats (n = 18). Systolic blood pressure was measured by the tail-cuff method prior to STZ injection and then weekly. After 3 months, plasma creatinine, and 24-hour urine albumin and creatinine were measured and kidneys harvested for morphometry. RESULTS: Ang II infusion increased systolic blood pressure in diabetic and ND rats. When combined with diabetes, Ang II increased albumin excretion rate (14-fold, p < 0.05), plasma creatinine (1.5-fold, p < 0.005) worsened creatinine clearance (37%, p < 0.002) and increased glomerular basement membrane width (1.2-fold, p < 0.0001). CONCLUSION: Ang II caused moderate hypertension and accelerated diabetic nephropathy and glomerular structural changes. The Ang II-infused STZ-diabetic rat is an excellent model to study the deleterious glomerular effects of hypertension on diabetes independent of genetic traits. PMID- 15539792 TI - Molecular mechanism of oxalate-induced free radical production and glutathione redox imbalance in renal epithelial cells: effect of antioxidants. AB - BACKGROUND: Peroxidation of renal cells is a critical event in the nucleation and formation of calcium oxalate crystals under hyperoxaluric conditions. We previously demonstrated that oxalate-induced peroxidative injury is one of the major mechanisms in promoting crystal attachment to renal epithelial cells. METHODS: In this study we have demonstrated that the mechanism of oxalate-induced peroxidative injury is through the induction of TGF-beta1 and glutathione (GSH) redox imbalance in LLC-PK1 cells. RESULTS: LLC-PK1, renal epithelial cells exposed to oxalate had significantly higher reactive oxygen species (ROS) production; higher TGF-beta1 levels, as measured by ELISA (1.89 +/- 0.035 fold increase) or Western blot (1.65 +/- 0.01 fold increase); increased malondialdehyde formation; increased LDH release, and loss of cell viability. In addition, oxalate exposure significantly decreased GSH content, glutathione reductase, glucose-6-phosphate dehydrogenase activities, and increased oxidized GSH content. Treatment with vitamin E, neutralizing anti-TGF-beta antibody, or diphenylene iodium, an inhibitor of NAD(P)H oxidase, significantly inhibited oxalate-induced ROS production and prevented peroxidative injury and cytolysis. Vitamin E, catalase, or desferoxamine treatment also significantly restored the oxalate-induced cellular GSH redox status toward the control level, and vitamin E treatment significantly attenuated the oxalate-mediated increase in TGF-beta1 protein in cultured LLC-PK1 cells. CONCLUSIONS: This is the first study to demonstrate that the mechanism of oxalate-induced free radical production in renal tubular epithelial cells is through the activation of NAD(P)H oxidase via cytokine TGF-beta1 induction. These results also provide direct evidence that antioxidant therapy might prevent calcium oxalate nucleation and kidney stone formation by preventing oxalate-mediated peroxidative injury and GSH redox imbalance. PMID- 15539793 TI - Novel insights into the aetiology and pathogenesis of hypopituitarism. AB - Recent advances in our knowledge of pituitary development, acquired mainly from animal models, have enhanced our understanding of the aetiology of isolated growth hormone deficiency (IGHD) and combined pituitary hormone deficiency (CPHD), as well as several syndromic forms of growth hormone deficiency (GHD). A number of developmental genes known to be important for organ commitment and cell differentiation and proliferation (HESX1, LHX3, LHX4, PROP1 and PIT1) have been implicated in CPHD with or without other syndromic features. Phenotypes associated with these genetic mutations and their inheritance may be highly variable. Functional analyses of these mutations reveal valuable insights into the function of the proteins and hence into the effect of these mutations on phenotype. Novel insights have been gained into the mechanisms whereby these genes are associated with particular phenotypes as a result of murine transgenesis, e.g. type II autosomal dominant GHD. Mutations within known genes account for a small proportion of cases of IGHD and CPHD, suggesting the role of other as yet unidentified genetic and environmental factors. Hence, genetic testing will in the future have a greater role to play in understanding the mechanisms leading to particular hypopituitary phenotypes and also in predicting the evolution of these disorders. There is, however, no substitute for careful delineation of the phenotype prior to undertaking genetic studies. PMID- 15539794 TI - Molecular mechanisms of thyroid dysgenesis. AB - Thyroid dysgenesis (TD) is the most prevalent form of congenital hypothyroidism. Ttf-1, Ttf-2, Pax8 and the Tshr are expressed at early stages of thyroid development and are implicated in thyroid ontogeny. Mutations in these genes have been found in some cases of TD. The prevalence of familial forms of TD is significantly higher than expected if the disease was only sporadic, allowing to postulate a genetic basis of the disease. Linkage analysis and mutational screening of the four above-mentioned genes in familial forms of TD showed their exclusion as contributors to the disease in some families, implicating genetic heterogeneity and involving other genetic mechanisms. Strategies to uncover new genes involved in TD are therefore needed. We underscore differences in the temporal expression patterns during the human thyroid development with those in animal models. Further, the extrathyroid expression of these genes during human development enables to define the gene-specific malformations that may be present in patients bearing mutations. The data gathered on molecular thyroid development enable precise genetic counselling of affected families. By increasing our knowledge of thyroid development, we hope to uncover new perspectives of genetic screening and eventually of early in utero treatment. PMID- 15539795 TI - The adrenal. AB - During the past 15 years, considerable progress has been made in our understanding of the genetic basis of adrenal development and function. More than 30 single gene disorders have now been identified that can affect the hypothalamic-pituitary-adrenal axis in humans (fig. 1, 2; table 1). This review highlights recent advances in the molecular pathology of: (1) adrenal hypoplasia, (2) adrenal destruction, (3) disorders of adrenal steroidogenesis, (4) adrenal steroid resistance and (5) activation of the adrenal axis/tumorigenesis. Characterizing the molecular basis and natural history of these conditions is providing fascinating insight into adrenal development and function and can help to focus treatment and counselling of patients appropriately. However, ongoing translation of research findings into clinical practice is needed if patient care is to be influenced significantly. PMID- 15539796 TI - Growth hormone treatment: cancer risk. AB - There have been concerns that growth hormone (GH) therapy may be associated with an increased risk of cancer. Although data are limited and conflicting, one recent report on cancer risk in individuals with no cancer history or risk factors for cancer who were treated with pituitary GH demonstrated a small increased risk of colon cancer and deaths from colon cancer and Hodgkin disease. The data from cancer survivors have consistently shown no increased risk of recurrence of the primary tumor in survivors of all tumor types who are treated with GH. One recent study did show a small increased risk of second solid tumors in survivors previously treated with GH. Limited data suggest that GH therapy is not associated with excess cancer risk in individuals with Langerhans cell histiocytosis and neurofibromatosis type 1. Overall, the clinical data are reassuring, but continued surveillance is mandatory. PMID- 15539797 TI - Impact of growth hormone status on body composition and the skeleton. AB - Severe growth hormone (GH) deficiency (GHD) induces a well-defined clinical entity encompassing, amongst the most reported features, abnormalities of body composition, in particular increased fat mass, especially truncal, and reduced lean body mass. The results from virtually all treatment studies are in agreement that GH replacement improves the body composition profile of GHD patients by increasing lean body mass and reducing fat mass. More recently, the observations have been extended to adults with partial GHD, defined by a peak GH response to insulin-induced hypoglycaemia of 3-7 microg/l. These patients exhibit abnormalities of body composition similar in nature to those described in adults with severe GHD; these include an increase in total fat mass of around 3.5 kg and a reduction of lean body mass of around 5.5 kg. The increase in fat mass is predominantly distributed within the trunk. The degree of abnormality of body composition is intermediate between that of healthy subjects and that of adults with GHD. The impact of GH replacement on body composition in adults with GH insufficiency, although predictable, has not been formally documented. The skeleton is another biological endpoint affected by GH status: in adults with severe GHD, low bone mass has been reported using dual energy x-ray absorptiometry (DEXA) and other quantitative methodologies. The importance of low bone mass, in any clinical setting, is as a surrogate marker for the future risk of fracture. Several retrospective studies have documented an increased prevalence of fractures in untreated GHD adults. Hypopituitary adults with severe GHD have reduced markers of bone turnover which normalize with GH replacement, indicating that GH, directly or via induction of insulin-like growth factor-I, is intimately involved in skeletal modelling. Whilst the evidence that GH plays an important role in the acquisition of bone mass during adolescence and early adult life is impressive, the impact of GHD acquired later in adulthood is less clear. Recently we examined the relationship between bone mineral density (BMD) and age in 125 untreated adults with severe GHD using DEXA. A significant positive correlation was observed between BMD (z-scores) and age at all skeletal sites studied. Overall, few patients, except those aged less than 30 years, had significantly reduced bone mass (i.e. a BMD z-score of less than -2); correction of BMD to provide a pseudo-volumetric measure of BMD suggested that reduced stature of the younger patients may explain, at least in part, this higher frequency of subnormal BMD z-scores. Despite normal BMD, however, an increase in fracture prevalence may still be observed in elderly GHD adults as a consequence of increased falls related to muscle weakness and visual field defects. PMID- 15539798 TI - Cardiovascular effects of growth hormone treatment: potential risks and benefits. AB - Growth hormone (GH) and insulin-like growth factor-I are involved in heart development and in maintaining cardiac structure and performance. Cardiovascular disease has been reported to reduce life expectancy both in GH deficiency (GHD) and in GH excess. Patients with GHD suffer from abnormalities of left ventricular performance, i.e. reduced diastolic filling and impaired response to peak exercise. Patients with GHD also have increased intima-media thickness at the common carotid arteries, associated with a higher occurrence of atherosclerotic plaques, which may further aggravate the haemodynamic conditions. This may contribute to increased cardiovascular and cerebrovascular risk. These cardiovascular abnormalities can be reversed, at least partially, with GH replacement therapy. In recent years, GH therapy has been used to increase cardiac mass in ischaemic or dilated cardiomyopathy, but the results have produced contradictory data. PMID- 15539799 TI - Growth hormone and glucose homeostasis. AB - Patients with active acromegaly are insulin-resistant and glucose-intolerant, whereas children with growth hormone (GH) deficiency (GHD) are insulin-sensitive and may develop fasting hypoglycaemia. Surprisingly, however, hypopituitary adults with unsubstituted GHD tend to be insulin-resistant, which may worsen during GH substitution. During fasting, which may be considered the natural domain for the metabolic effects of GH, the induction of insulin resistance by GH is associated with enhanced lipid oxidation and protein conservation. In this particular context, insulin resistance appears to constitute a favourable metabolic adaptation. The problem is that GH substitution results in elevated circadian GH levels in non-fasting patients. The best way to address this challenge is to employ evening administration of GH and to tailor the dose. Insulin therapy may cause hypoglycaemia and GH substitution may cause hyperglycaemia. Such untoward effects should be minimized by carefully monitoring the individual patient. PMID- 15539800 TI - Genetics and variation in phenotype in Noonan syndrome. AB - Noonan syndrome is a well-known clinical entity comprising multiple congenital anomalies characterized by typical facial features, short stature and congenital heart defect. Approximately 50% of cases are sporadic. Familial cases are generally autosomal dominant. In 2001 a gene responsible for Noonan syndrome, PTPN11, encoding for the non-receptor protein tyrosine phosphatase SHP-2, was identified. Mutation analysis of the PTPN11 gene was carried out in Nijmegen in 150 patients with Noonan syndrome. Mutations were found in 68 patients (45%), the most common being A922G in exon 8. In exon 4 a mutation was found that encoded the C-SH2 domain of the PTPN11 gene in two unique patients who shared some uncommon features. A 218C-->T mutation was found in exon 3 in one patient with Noonan syndrome and mild juvenile myelomonocytic leukaemia. PMID- 15539801 TI - Molecular basis of Sotos syndrome. AB - This paper describes the isolation of a novel human gene, NSD1, from the 5q35 breakpoint of t(5;8)(q35; q24.1) in a patient with Sotos syndrome, and NSD1 mutation analysis. Of 112 (95 Japanese and 17 non-Japanese) patients analyzed, 16 (14%) had a heterozygous NSD1 point mutation (10 protein truncation types and six missense types) and 50 (45%) a approximately 0.7-Mb microdeletion involving NSD1. The results indicated that haploinsufficiency of NSD1 is the major cause of Sotos syndrome, and NSD1 plays a role in growth and brain development in humans. Detailed clinical examinations provided a genotype-phenotype correlation in Sotos syndrome, i.e. in patients with deletions, overgrowth is less obvious and mental retardation is more severe than in those with point mutations, and major anomalies were exclusively seen in the former. The results also indicated that Sotos syndrome due to a deletion falls into a contiguous gene syndrome, while Sotos syndrome due to an NSD1 point mutation is a single gene defect, occasionally with an autosomal dominant mode of inheritance. The genomic structure around the deleted and flanking regions revealed the presence of two sets of low copy repeats through which the microdeletion in Sotos syndrome is mediated. PMID- 15539802 TI - Proceedings of the SGA Launch Symposium and the 15th Novo Nordisk Symposium on Growth Hormone and Endocrinology. April 1-3, 2004. Malta. PMID- 15539803 TI - Beta-cells in type 2 diabetes: a loss of function and mass. AB - Type 2 diabetes mellitus manifests itself in individuals who lose the ability to produce sufficient amounts of insulin to maintain normoglycaemia in the face of insulin resistance. The ability to secrete adequate amounts of insulin depends on beta-cell function and mass. Chronic hyperglycaemia is detrimental to pancreatic beta-cells, causing impaired insulin secretion and playing an essential role in the regulation of beta-cell turnover. This paper will address the effect of chronically elevated glucose levels on beta-cell turnover and function. In previous studies we have shown that elevated glucose concentrations induce apoptosis in human beta-cells due to an interaction between constitutively expressed Fas ligand and upregulated Fas. Human beta-cells produce interleukin (IL)-1beta in response to high glucose concentrations, independently of an immune mediated process. This was antagonized by the IL-1 receptor antagonist (IL-1Ra), a naturally occurring anti-inflammatory cytokine also found in the beta-cell. Therefore the balance of IL-1beta and IL-1Ra may play a crucial role in the pathogenesis of diabetes. Inhibition of glucotoxicity represents a promising therapeutic stratagem in diabetes therapy to preserve functional beta-cell mass. PMID- 15539804 TI - Pitfalls in the diagnosis of acromegaly. AB - Acromegaly is a disfiguring and disabling illness which, when inadequately treated, reduces life expectancy. An implication of the ability to offer effective treatment is the increased onus on physicians of all sorts to ensure acromegaly is diagnosed and treated as early as possible. To this end, criteria for the diagnosis of acromegaly have been proposed in the consensus statement of Giustina et al. However, other data suggest that the proposed criteria are not rigorous enough and strict adherence to the guidelines would result in failure to diagnose a significant number of patients. A review of published experience suggests that the combination of a GH nadir during an oral glucose tolerance test of <0.25 microg/l plus a normal age-related insulin growth factor-I level makes the diagnosis of acromegaly extremely unlikely. PMID- 15539805 TI - Treatment of acromegaly. AB - Mortality is increased in individuals with acromegaly unless serum growth hormone (GH) levels are below 2 microg/l and serum insulin-like growth factor (IGF)-I levels are normal following treatment. These combined criteria have been used to define remission of the disorder in this review. Transsphenoidal surgery achieves remission targets in an average of 55% of patients. For those not in remission following surgery, options include repeat surgery or use of adjuvant therapy. Fractionated external beam pituitary radiotherapy achieves 10-year remission rates of 47% but leaves patients exposed to excess GH until remission occurs. Stereotactic radiotherapy and gamma knife radiosurgery achieve remission rates of 40% over 3 years, and dopamine agonists produce remission in about 20% of patients. Somatostatin analogues induce remission in 59% of patients within the first year of treatment. The GH receptor antagonist pegvisomant leads to remission in 90% of patients, using IGF-I levels for assessment. Optimal treatment for a patient with acromegaly thus depends on the likely efficacy of treatment, cost, surgical skill, severity of side effects, tolerability, control of tumour growth, and improvement in complications related to tumour mass. A primary surgical approach, followed by medical therapy for those not in remission, remains the preferred option in most centres. PMID- 15539806 TI - Optimizing growth hormone efficacy: an evidence-based analysis. AB - For almost two decades, recombinant growth hormone (GH) has been in abundant supply to treat GH-deficient subjects to final height, yet the results continue to be suboptimal. Although there are numerous potential causes for this underachievement, the issues of poor compliance, ineffective dosing and lack of efficacy monitoring have been frequently promulgated to explain the phenomenon. Compliance is tied to knowledge, which is best promoted at the time of diagnosis by hospital-based personnel. GH pens have improved acceptability, but data on improvement in compliance are lacking. No GH device or delivery system has yet been shown to increase final height. Daily (or six times per week) dosing is clearly more effective, but increasing GH dose, especially during puberty, has a tendency to advance bone age and pubertal progression, which mitigates many of the effects. Finally, titration of dose to the insulin-like growth factor (IGF)-I level may prove to be useful in promoting final height; however, the converse is not true, i.e. increments in IGF-I level have not been shown to correlate with increments in height velocity. Thus, these issues with respect to achievement of final height are far from solved. PMID- 15539807 TI - Liquid growth hormone: preservatives and buffers. AB - Growth hormone (GH) treatment is a successful medical therapy for children and adults with GH deficiency as well as for growth retardation due to chronic renal disease, Turner syndrome and in children born small for gestational age. For all of these conditions, treatment is long term and patients receive daily subcutaneous injections of GH for many years. Patient compliance is therefore of critical importance to ensure treatment benefit. One of the major factors influencing compliance is injection pain. Besides the injection device used, pain perception and local tissue reaction following injection are dependent on the preservative used in the formulation and the concentration of GH. Injection pain may also be related to the buffer substance and injection volume. A liquid formulation of GH, Norditropi SimpleXx, has been developed that dispenses with the need for reconstitution before administration. The formulation uses phenol (3 mg/ml) as a preservative (to protect product from microbial degradation or contamination) and histidine as a buffer. Alternative preservatives used in other GH formulations include m-cresol (9 mg/ml) and benzyl alcohol (3-9 mg/ml). Buffering agents include citrate and phosphate. Phenol has been successfully used as a preservative in drug formulations for more than 50 years and is considered a safe and effective agent which complies with strict international requirements for preservatives in drug formulations. In toxicological studies, no or only mild local reactions have been observed following subcutaneous administration of phenol (7.5 mg/ml), m-cresol (3-4 mg/ml) and benzyl alcohol (9 mg/ml). No general toxicity reactions were observed after subcutaneous administration of these agents. Clinical evaluation of the preservatives and buffers used in Norditropin SimpleXx showed that pain perception was similar between formulations containing phenol and benzyl alcohol, whereas m-cresol was associated with more painful injections than benzyl alcohol. Furthermore, patients reported more pain following injection of a citrate-buffered solution than after a histidine buffered solution. More pain was also reported following large volume injections and following injections with solutions containing high protein concentrations. In summary, optimization of the preservative and buffer content of a liquid GH formulation may reduce injection pain and lead to improved patient compliance. PMID- 15539808 TI - Igf-I and longevity. AB - Recent animal studies have demonstrated evidence of the involvement of insulin and insulin-like growth factor (IGF)-I signalling in the control of ageing and longevity. Disruption of insulin/IGF-I signalling pathways significantly extends lifespan in several animal models. Similarities among these signalling pathways in animals and humans raise the possibility that modifications in the IGF-I signalling system could also extend lifespan in humans. However, in contrast to the findings in animal studies, reduced IGF-I activity in humans is not associated with longevity. In humans, low IGF-I activity is even associated with an increased risk of developing cardiovascular disease and diabetes. High IGF-I activity in humans is associated with an increased risk of developing cancer. In addition, genetic predisposition and lifestyle play a major role in determining age-associated disease. For each individual there is probably a specific optimal 'setpoint' for the insulin/growth hormone/IGF-I axis which co-determines survival. PMID- 15539809 TI - Adrenopause. AB - Dehydroepiandrosterone(DHEA) and DHEA-S are steroids that are abundantly produced by the adrenal gland. Plasma concentrations of DHEA and DHEA-S increase during adrenarche but decrease steadily after puberty. Although DHEA and DHEA-S have few intrinsic androgenic actions, they have recently attracted widespread attention due to their beneficial anti-aging effects. We clarified the beneficial effects of DHEA as an anti-aging steroid with regard to its stimulation of the immune system and its anti-diabetes, anti-atherosclerosis, anti-dementia (neurosteroid), anti-obesity and anti-osteoporosis effects. There are two possible biochemical and molecular mechanisms: direct action via the DHEA receptor on the target gene; and indirect action. We identified a high affinity of DHEA binding in human T lymphocytes by searching for the target genes that are induced in activated T lymphocytes in the presence of DHEA, determined the gene sequence and named DHEA induced dual p38-specific phosphatase (DDSP). DDSP transgenic mice have been created to identify the anti-aging effects of DDSP. The conversion of DHEA to estrone by cytochrome P450 aromatase in primary cultured human osteoblasts was clarified. We are currently undertaking an open trial of DHEA replacement therapy. PMID- 15539812 TI - Akting and cycling: a tale of the pituitary. AB - Pituitary tumours are characterized by a series of phenotypic abnormalities, but the molecular nature of the underlying defects has proved peculiarly intractable. Oncogenes and tumour suppressor genes involved in other tumours do not appear to play a major role in the pathogenesis of pituitary tumours. In addition, germline genetic disorders in which pituitary tumours are a common feature have not shed much light on the more common sporadic tumour. A number of defects in specific feedback regulation in the secretory tumours have been identified, but it is presently unclear as to what extent these are a consequence of the tumour, possibly enhancing its growth or survival, rather than the cause. However, recent studies on the cell cycle have demonstrated significant abnormalities that have been traced to a cytoplasmic kinase which appears to be abnormally expressed in the majority of pituitary adenomas, and we are beginning to see a possible unifying abnormality. PMID- 15539813 TI - Growth hormone treatment of short children born small for gestational age: a US perspective. AB - Research during the last decade shows clearly that growth hormone (GH) therapy causes a sustained increase in growth velocity when applied to short children born small for gestational age (SGA). This occurs even though GH deficiency per se is an unlikely explanation for their lack of catch-up growth. In the United States, children born weighing less than -2 SD for gestational age and who show no growth recovery (usually defined as stature persisting below -2 SD at age 2 years) are eligible for GH treatment using doses up to 0.48 mg/kg per week. The management of these children brings new challenges to the pediatric endocrinologist. Intrauterine growth retardation reflects a variety of etiologies, some of which merit special consideration and may respond variably to GH. The dose of GH used exceeds physiologic replacement and is higher than that commonly used to treat other non-GH-deficient conditions such as Turner syndrome. Thus, what constitutes optimal therapy in terms of dose, timing and patient selection remains an important question. While GH therapy provides a means by which one aspect of the SGA syndrome can be helped, there are other issues for SGA apart from height. Future efforts should include studies that better define how GH should be used in the short child born SGA and address more broadly the medical, social and psychological needs of these patients. PMID- 15539814 TI - Growth hormone treatment in short Japanese children born small for gestational age. AB - Recent reports have shown that high-dose growth hormone (GH) treatment in short children born with small for gestational age (SGA) resulted in a pronounced acceleration of linear growth. We describe the results of multicenter trials of recombinant human GH (rhGH) treatment in short SGA children in Japan. Two clinical studies were performed and the results were combined. Study 1 comprised 104 SGA children and study 2 comprised 61 SGA children. The patients were divided into three groups: group 1 consisted of 20 patients (13 boys and 7 girls) who received rhGH 25 microg/kg per day six or seven times per week in the first year and 50 microg/kg per day in the second year and thereafter; group 2 consisted of 48 patients (28 boys, 20 girls) who received rhGH 45/50 microg/kg per day; group 3 consisted of 44 patients (28 boys, 16 girls) who received 90/100 microg/kg per day. The mean increments in height SDS were 0.46, 0.67 and 0.94 SD in boys and 0.49, 0.79 and 0.93 SD in girls in groups 1, 2 and 3, respectively. The mean increment in height SDS at 2 years in group 3 was significantly greater than that in group 1, but it was not significantly different from that in group 2 in boys and girls. Our data demonstrated that high-dose GH administration significantly improved height velocity and height SDS in short SGA children. Additional studies are necessary to optimize a long-term GH treatment regimen and combined luteinizing hormone releasing hormone analog treatment for final height. Careful observation is also necessary to assess the metabolic effects of high-dose GH, especially on carbohydrate metabolism. PMID- 15539815 TI - Growth hormone treatment strategy for short children born small for gestational age. AB - Several studies performed in the last 15 years have shown that growth hormone (GH) induces a profound catch-up in height in short children born small for gestational age (SGA). We know from more recent studies that final height can be normalized through GH treatment. In Europe, GH is now a recognized indication, enabling treatment of short children born SGA. Treatment is given to the most severe growth-retarded children after the age of 4 years. A dose of 0.035 mg/kg per day is recommended. However, in our opinion a higher dose would be more efficient in very short children, especially if they are treated later in childhood. PMID- 15539817 TI - Adipose tissue magnetic resonance imaging in the newborn. AB - Infancy is a period of rapid adipose tissue accumulation, and influences during early development are plausible determinants of altered adiposity. The distribution, as well as the quantity of adipose tissue, is a marker of health and disease. Previous methods for the assessment of body composition in infants have been indirect and thus unable to determine adipose quantity reliably, nor assess adipose tissue distribution. Adipose tissue magnetic resonance imaging is direct, non-invasive, radiation free and suitable for serial examinations in infancy. Adipose tissue depots are quantified individually and summated to provide an accurate measure of depot-specific and total adiposity. We have adapted this technique for application to newborns and, to date, have imaged over 100 term and preterm infants. PMID- 15539818 TI - Efficacy and safety of long-term continuous growth hormone treatment of children born small for gestational age. AB - Twelve years of growth hormone (GH) therapy of short children born small for gestational age (SGA) have demonstrated that GH is an effective and well tolerated therapy. Most children will reach a normal adult height (AH). AH of 55 SGA adolescents was comparable for those treated with a GH dose of 1 or 2 mg/m2 (approximately 0.033 or 0.066 mg/kg) per day, mean (SD) AH SDS being -1.2 (0.7) and -0.8 (0.7), respectively. GH therapy had no influence on the age at onset, the progression of puberty, duration of puberty and pubertal height gain. GH therapy induced higher fasting and glucose-stimulated insulin levels after 1 and 6 years, but 6 months after GH stop, all levels returned to normal. At baseline mean systolic blood pressure was significantly increased, but both systolic and diastolic blood pressure decreased significantly during 6 years of GH and remained so after GH stop. GH therapy demonstrated a beneficial effect on serum lipid profiles, body composition, bone mineral density and head growth. Treatment with 2 mg GH/m2 per day induced mean serum IGF-I levels of +2 SDS, whereas IGF-I levels remained within the normal range with 1 mg GH/m2 per day. In conclusion, long-term GH therapy of short SGA children with 1 mg/m2 per day appears to be effective and safe. Since the future consequences of high serum IGF-I levels during long-term GH therapy with 2 mg/m2 per day are as yet unknown, it seems safer to treat short prepubertal SGA children with a GH dose of 1 mg/m2 per day when children are to be treated continuously for many years. PMID- 15539819 TI - Genetic services in Latin America. AB - This special issue of Community Genetics reviews some of the most important developments in medical genetics in key countries of Latin America. Contributions to this issue were prepared for a special consultation of the World Health Organization held in Porto Alegre, Brazil, on June 19, 2003. Latin America is a region of medium- to low-income countries characterized by socioeconomic problems, with large segments of the population living in poverty and extreme disparities in the distribution of wealth. A rise in chronic diseases typical of the processes of industrialization and urbanization coexists with the persistence of nutritional and infectious diseases characteristic of poverty and underdevelopment. Over the last 2 decades of the 20th century, birth defects and genetic disorders have increased their share of morbidity and mortality, and tertiary-care-based genetic services have developed in urban areas. Although privatization of health care is eroding the public sector, the public institutions continue to be the main providers of genetic services for the bulk of the population and the leaders in research. The development of clinical genetics in the region is concentrated in tertiary-care centers in large cities, although a recent trend began extending genetic services to the community. PMID- 15539820 TI - The role of the World Health Organization in promoting medical genetics in Latin America. AB - The Human Genetics Program of the World Health Organization (WHO) has analyzed the needs for genetic services worldwide and the promotion of genetic approaches for the prevention of diseases linked to primary health care. This article presents a summary of the most important initiatives of the WHO in this field, which have served as a background for the Consultation in Medical Genetics in Latin America, which took place in Porto Alegre, Brazil, in June 19, 2003, and whose working reports are published in this special issue of Community Genetics. PMID- 15539821 TI - The Latin-American network of human genetics. AB - The Latin-American Network of Human Genetics (RELAGH) is an internet-based federation of human and medical genetic societies of Latin America constituted in 2001. Member societies include the Mexican Association of Human Genetics, the Brazilian Society of Clinical Genetics, the Human Genetics Section of the Argentinean Society of Genetics, the Colombian Society of Human Genetics and the Panamanian Human Genetics Society. RELAGH is a full member of the steering committee of the International Federation of Human Genetics Societies. A core activity of RELAGH is to become a platform for the integration and collaboration of human geneticists across Latin America regarding training, services and research in medical genetics in the region. PMID- 15539822 TI - ECLAMC: the Latin-American collaborative study of congenital malformations. AB - DEFINITION: ECLAMC ('Estudio Colaborativo Latino Americano de Malformaciones Congenitas') is a program for the clinical and epidemiological investigation of risk factors in the etiology of congenital anomalies in Latin-American hospitals, using a case-control methodological approach. It is a voluntary agreement among professionals lacking institutional base as well as designated budgets. ECLAMC has been usually funded by research-funding agencies rather than public health ministries. The National Research Councils of Argentina and Brazil have been the main sources of support during its 36 years of existence. Since vital and health statistics are unreliable in South America, ECLAMC collects all the information required for the denominators in a hospital-based sample of births. ECLAMC can be defined as a continental network of persons interested in research and prevention of birth defects. HISTORY AND EVOLUTION: From the institutional point of view, ECLAMC has had headquarters in diverse centers of Argentina and Brazil, but always as an independent research project, without a defined administrative link. ECLAMC began operating in 1967, as an investigation limited to the city of Buenos Aires, Argentina, and it gradually expanded until covering all the 10 countries of South America as well as Costa Rica and the Dominican Republic. Even though ECLAMC has maintained essentially the same original experimental design since 1967, due to the data accumulated by the program, the increasing experience as well as the development in science, technical modifications occurred including a DNA bank and a fully informatized data handling system. Since 1974 ECLAMC has been a founder member of the International Clearinghouse for Birth Defects Monitoring Systems; since 1994 a WHO Collaborating Center for the Prevention of Congenital Malformations, and since 2000 a collaborating member of the NIH Global Netwok for Women's and Children's Health Research. METHODOLOGY: The maternity hospital network of ECLAMC examines around 200,000 births per year. All major and minor anomalies diagnosed at birth in infants weighing 500 g or more are registered according to a manual of procedures. The next non-malformed baby of the same sex born in the same hospital is selected as a control subject for each case. Thus, a one-to-one healthy control group matched by sex, time and place of birth is obtained. As a system of epidemic surveillance, ECLAMC systematically observes the fluctuations in the frequencies of different malformations and, in the case of an alarm for a probable epidemic of a given malformation, at a given moment, and given area, it acts to identify its cause. As termination of pregnancy has severe legal restrictions in South America, prevention of birth defects should concentrate on primary, preconceptional and tertiary measures. Tertiary measures aim to avoid complications of the affected patients from the medical, psychological, and social standpoints. PMID- 15539823 TI - Clinical genetics in developing countries: the case of Brazil. AB - There are many impediments to the progress of clinical and medical genetics in developing countries. Higher priorities concerning basic health care usually take precedence over genetic diseases and birth defects among medical professionals and public health officials. This is so in spite of the fact that the global prevalence of these conditions seems higher than in the developed world and that limited resources enhance the burden on individuals, families and populations. Furthermore, as a consequence of recent advances in medical genetics, demand for genetic services has increased, reinforcing the need for programs for the management and prevention of genetic diseases and birth defects, especially at primary health care level. An overview of these issues in Brazil is presented here, with information on the health system, the evolution of medical and clinical genetics in the country, and the situation of medical and clinical genetic services. We discuss proposals for implementing appropriate, ethically acceptable and equitable clinical genetic services for the Brazilian population. PMID- 15539824 TI - Medical genetics services in the city of Sao Paulo, Brazil. AB - The city of Sao Paulo is located in the center of a metropolitan area with nearly 18 million inhabitants and 300,000 births/year. The currently existing medical genetics services are unable to meet the demand, due to their insufficient physical and personnel infrastructure. Institutions and experts in medical genetics could give short training and refresher courses to health professionals to enable them to work in the public health network. The city has a reasonably well developed health care network, represented by the Single Health System (Sistema Unico de Saude - SUS) and by the Family Health Program (Programa de Saude da Familia - PSF). The financial resources for such actions originate in the budget of the managing agencies of such systems. The limitations of genetic services provided to the population of the city could be overcome in a short period of time by developing programs within the public health care network. The city has institutions, professionals and financial resources to make this project feasible. To that end, the competent authorities of the Sao Paulo State and City Secretariats of Health should take managerial responsibility for the genetic services in the city. PMID- 15539825 TI - Medical genetic services in the state of Rio de Janeiro, Brazil. AB - Rio de Janeiro is a state with close to 15 million inhabitants and approximately 250,000 births per year. The state counts nine clinical genetic units in public institutions, providing for 9,400 outpatient consultations yearly, which is insufficient to cover the estimated needs. Laboratory tests such as cytogenetics, inborn errors of metabolism and molecular studies are available on a limited basis. Newborn screening in the public health system is being performed for phenylketonuria, congenital hypothyroidism and sickle cell disease. In the state there are also special treatment programs for osteogenesis imperfecta and Gaucher's disease, subsidized by the Brazilian Ministry of Health. Presently, efforts of medical geneticists are concentrated on highlighting the practical relevance of clinical genetics, and the need to integrate the specialty into the public health system in a functional network of genetic services. PMID- 15539826 TI - Genetic services and research in the state of Minas Gerais, Brazil. AB - The state of Minas Gerais in Brazil has a surface of 586,528 km(2), and 18 million inhabitants. Infant mortality rate is 20/1,000, and congenital anomalies are its second cause. There are 11 medical schools where basic genetics, but not clinical genetics, is taught. Genetic services in the state include: newborn screening for hypothyroidism, phenylketonuria, sickle cell disease and cystic fibrosis; clinical-genetic diagnostic evaluation and counseling; prenatal diagnosis, fetal medicine and paternity testing. Medical genetic services and research are underdeveloped because of limitations such as lack of health policies in genetics, small number of trained specialists, little knowledge about genetics among health professionals and low reimbursement rates. PMID- 15539827 TI - Genetic services in Chile. AB - Demographic changes in Chile have positioned congenital malformations as a major cause of infant morbidity and mortality. At the same time, medical genetics has become increasingly important in relation to the diagnosis and management of individuals with birth defects and hereditary conditions as well as in the study of pathological pregnancies and reproductive problems. In addition, recent advances in genomic research are expanding the relevance of medical genetics to medicine as a whole. This article reviews the clinical genetic resources currently available in Chile; the teaching of genetics in undergraduate, graduate, and continued medical education; some relevant interventions that have taken place in our country, e.g. the expansion of the newborn screening program and the initiation of a folic acid fortification program, and recent efforts to enhance population access to clinical genetics services. PMID- 15539828 TI - Genetic services in Colombia. AB - Medical genetic services, including clinical genetics, cytogenetics, biochemical and molecular genetics and paternity testing, are performed in Colombia in the more developed medical schools or university institutions, in nine major cities of the country. Accessibility to genetic services is limited by medical care reimbursement laws which do not cover clinical genetic services nor genetic tests. Paternity testing is performed free of charge by a governmental welfare institution, if a legal claim is made against an alleged father. Basic teaching of genetics in medical schools is mandatory, but is very uneven and limited to the better schools. Postgraduate medical genetic training is offered by four different programs of similar quality. Research is performed on some of the most prevalent genetic conditions and on population genetic issues. PMID- 15539829 TI - Community-based program for the diagnosis and prevention of genetic disorders in Cuba. Twenty years of experience. AB - The author's experience of 20 years as director of the medical genetic services program in Cuba is presented. The setting of the infrastructure for equipment and the training of personnel for the medical genetic program began in 1981 in the city of Havana, and was progressively extended to cover the whole country in 1988. Between 1982 and 2002, 2.8 million pregnant women were tested for sickle cell carrier status, 96,000 carriers and 4,786 couples at risk were detected and offered genetic counseling and prenatal diagnosis. In the same period, the combination of maternal serum alpha-fetoprotein screening and fetal ultrasound led to the prenatal diagnosis of anomalies in several thousand fetuses. The accessibility to legal abortion, and the autonomous decisions by the majority of couples to terminate abnormal pregnancies reduced the prevalence of neural tube defects and sickle cell disease at birth by 90 and 65%, respectively, by 2002. In the span of approximately 20 years, 22,690 pregnant women at risk received prenatal chromosomal diagnosis. Newborn screening for phenylketonuria and congenital hypothyroidism was established. Genetic counseling was offered to every detected person or family at risk for genetic conditions. The network of medical genetic services established in the country received a very positive acceptance by the population. A very successful connection with the primary health care level was established. PMID- 15539830 TI - Genetic services in Ecuador. AB - Ecuador has a population of 12 million inhabitants, composed of a variety of ethnic groups. The majority of the population lives in rural areas. There is a severe deficiency in health services, and most diseases are associated with poor life conditions: malnutrition, parasitic diseases and infections. At the same time, however, chronic diseases, e.g. cancer, have increased in prevalence. Most genetic services are provided by the private sector, which offers clinical genetic consultations and counseling, cytogenetic analyses, a limited number of molecular and biomedical tests, and prenatal diagnosis. Genetic research is centered in the universities of important cities (Quito, Guayaquil and Cuenca). The government sponsors some university research projects but does not conduct health actions in genetics in the community. There is an incipient interest in ethic and legal issues in genetics, such as how to provide modern genomic medicine in a resource-poor country. PMID- 15539831 TI - Genetic services in Mexico City. AB - In the year 2002, the population of Mexico was approaching 103 million inhabitants. Approximately 74% of them live in the cities with a continuous migration from rural to urban areas. Genetic departments are concentrated in the capital and other big cities. In this paper we review the current status of genetic departments in Mexico City, emphasizing the main areas of genetic services offered to the public and involved in research. We also comment on the deficiencies identified and suggest recommendations to improve the quality of the genetic services offered to the Mexican population. PMID- 15539832 TI - Medical genetics in Paraguay. AB - Paraguay is a developing country with low levels of health coverage, with 81% of the population without health insurance, a proportion that reaches 98.1% among the poor, 93% among the rural population and 91.7% among the mainly Guarani speaking population. The infant mortality rate is 19.4 per 1,000, although there is gross under-reporting. Maternal mortality rate is alarmingly high at 110.9 per 100,000 livebirths, reaching 420.5 in rural areas. There are only two clinical geneticists and four biochemists trained in human genetics, and virtually all genetic services in the country are concentrated in the 'Instituto de Investigaciones en Ciencias de la Salud' (IICS) from the National University. The teaching of medical genetics in medical schools is included in physiology and pathology courses, while at the postgraduate level, training in medical genetics is limited to pediatrics and gynecology. In 1999, a pilot newborn screening program was initiated to determine the frequency of congenital hypothyroidism and phenylketonuria and to provide early treatment for affected babies. Another pilot project recently launched by the Ministry of Health is the Program for the Prevention of Neural Tube Defects, mandating folic acid fortification of flour, but as of the end of 2003 it had not been implemented. Paraguay lacks adequate resources to provide accurate diagnoses and treatment of genetic conditions. PMID- 15539833 TI - Medical genetics in Peru. AB - Peru has a growing population characterized by notorious socioeconomic differences. The main health problems are acquired diseases related to sanitary conditions that affect mainly the large segment afflicted by poverty and extreme poverty. The state's health policy does not contemplate any action on congenital or genetic conditions, and genetic services are considered a very low priority. In spite of this, some of the best medical schools have undergraduate and graduate programs in genetics, and there is a growing group of specialists that makes its best to impulse the practice of medical genetics in Peru. Many ethical and legal dilemmas, common to other countries in the region, and derived from social inequality, and political, cultural and religious factors, are also faced in everyday practice. PMID- 15539834 TI - Medical genetics in Zulia, a State of Venezuela. AB - Zulia is a state located in the northwest of Venezuela. Congenital malformations, deformities and chromosomal anomalies are the second cause of infant and neonatal mortality. There are seven public and private groups providing genetic services, the most important of which, the Medical Genetic Unit at the Zulia University was created in 1973. So far, this unit has provided genetic services to 12,000 families, and has been responsible for undergraduate and postgraduate education in human and medical genetics. Prenatal diagnosis is performed at the Unit and a private practice group, the most frequent referral reason being advanced maternal age. The most frequent genetic diseases in the state are Huntington's disease, sickle cell anemia, neural tube defects and Down's syndrome. Research in genetics includes the clinical, epidemiological and molecular characterization of hereditary diseases, cancer, reproductive problems and genetic diversity. Other public groups are conducting research on dementias, including Alzheimer's disease, and on the genotoxic effects of environmental pollutants. PMID- 15539835 TI - Incidental prostatic cancer: repeat TURP or biopsy? AB - An incidental diagnosis of carcinoma is made in about 15% of patients undergoing transurethral or open surgery for prostatic adenoma. The importance of correct staging lies in the different clinical behaviors of the tumor according to the stage, which means that it will require different treatment. We present a review article on the diagnosis of residual neoplasia following transurethral resection of the prostate. PMID- 15539836 TI - Do we need to perform cystoscopy on all adults attending urology centres as outpatients? AB - INTRODUCTION: There has been considerable expansion in the use of flexible cystoscopy (FC) and people who can perform the procedure. Hence, there is a criticism that this procedure is being overused with no management benefit. MATERIALS AND METHODS: We audited the use of FC in a district hospital for a period of 1 year. The results of FC for non-standard indications (other than haematuria and check cystoscopy) were analysed for their diagnostic yield. RESULTS: Of the 1,390 FCs performed, 295 were done for non-standard indications. 46.14% of these cystoscopies had positive findings. Cancer detection rate was 6.10%. Cystoscopy altered the management in 14.08% of patients and was supportive to diagnosis and management in 32.06%. CONCLUSION: This procedure is certainly not overused and the ever-increasing requirement of this simple procedure has serious resource implications for the National Health Service. PMID- 15539837 TI - Comparison of WHO 1973, WHO/ISUP 1998, WHO 1999 grade and combined scoring systems in evaluation of bladder carcinoma. AB - OBJECTIVES: To compare WHO 1973, WHO/ISUP 1998 and WHO 1999 histologic grading systems, and also to evaluate the primary (most common) and secondary (second most common) patterns of cancer growth according to these three grading systems. MATERIAL AND METHODS: The study consisted of 87 bladder transurethral resections that were classified as grade 1, 2 and 3, and papillary urothelial neoplasm of low malignant potential (PUNLMP), low grade (LG) and high grade (HG) carcinoma considering WHO 1973 and WHO/ISUP, respectively. The WHO 1999 system was subdivided high grade into grades 2 and 3 (HG-2 and HG-3). For combined scoring, primary (most common) and secondary (second most common) grades according to extension were recorded for three grading systems. The number was repeated when only grade was seen in all extension of the tissue examined. A final combined score was obtained which ranged from 2 to 6 for the WHO 1973 and WHO/ISUP 1998 systems and from 2 to 8 for the WHO 1999 schema. The TNM system was used for the pathologic staging. RESULTS: When considering the pathological stage, there were statistical differences between the WHO 1973 grades (p=0.011 and p=0.000), and LG and HG carcinomas of WHO/ISUP 1998 (p=0.000) and also the WHO 1999 grades (p=0.010 and p=0.003), except PUNLMP. Regarding the combined scoring, significant differences were found between score 4 (2+2) and 5 (2+3) of WHO 1973 (p=0.014) and score 5 (LG+HG) and 6 (HG+HG) of WHO/ISUP 1998 (p=0.011). There was also a significant difference between scores 4 and 6, and 6 and 8 of the WHO 1999 combined scoring system (p=0.019 and p=0.019). WHO 1973, WHO/ISUP 1998 and WHO 1999 systems were positively correlated with the pathological stage (r(s)=0.30, r(s)=0.52 and r(s)=0.50, respectively), whereas there was weak association between the combined scoring systems and stage (r(s)=0.20, r(s)=0.18 and r(s)=0.19). Comparing these grading systems, the grade 2 of WHO 1973 was subdivided into LG and HG in WHO/ISUP 1998 and also LG-1and HG-2 in WHO 1999 systems. The group of HG carcinoma in WHO/ISUP 1998 which was subdivided into HG 2 and HG-3 in the WHO 1999 system was different statistically in relation to the stage. CONCLUSIONS: Our results revealed that the WHO 1999 system may be more useful to evaluate the bladder carcinoma histopathologically in comparison to the WHO 1973 and WHO/ISUP 1998 systems. PMID- 15539838 TI - Bladder tumor recurrence after primary surgery for transitional cell carcinoma of the upper urinary tract. AB - OBJECTIVE: Primary transitional cell carcinoma (TCC) of the upper urinary tract represents 6-8% of all TCC cases. Nephroureterectomy with removal of a bladder cuff is the treatment of choice. The rates of TCC recurrence in the bladder after primary upper urinary tract surgery described in the literature range between 12.5 and 37.5%. In a retrospective analysis we examined the occurrence of TCC after nephroureterectomy for upper tract TCC in patients without a previous history of bladder TCC at the time of surgery. METHODS: Between 1990 and 2002, 29 patients underwent primary nephroureterectomy for upper tract TCC. The mean age of the patients was 69.5 years. In 5 cases upper urinary tract tumors were multilocular, in the remaining cases unilocular in the renal pelvis (n=12) or the ureter (n=12). The follow-up was available for 29 patients with a mean follow-up of 3.37 (0.1-11.2) years. RESULTS: 11/29 (37.9%) patients had TCC recurrence with 9/11 patients having bladder TCC diagnosed within 2.5 years (0.9-6.0) after nephroureterectomy. 13/29 patients are alive without TCC recurrence, 3/29 patients died due to systemic TCC progression and 5/29 died of unrelated causes without evidence of TCC recurrence. CONCLUSION: Our data indicate a high incidence of bladder TCC after nephroureterectomy for primary upper tract TCC of up to 6 years after primary surgery. Because of the high incidence of bladder TCC within the first 3 years of surgery, careful follow-up is needed over at least this period. PMID- 15539839 TI - p53 Gene mutations in superficial bladder cancer. AB - OBJECTIVES: To assess the presence of p53 gene mutations in superficial tumors of the urinary bladder (transitional cell carcinoma) and their relationship to classic prognostic factors for cancer recurrence and progression. To analyze the implication of these mutations on the P53 protein structure. MATERIALS AND METHODS: Observational, cross-sectional study of 90 consecutive patients, 60 with superficial transitional cell carcinoma (pTa and pT1) and 30 without neoplastic disease (control group). Samples of bladder tumor and control normal mucosa were analyzed by polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) to detect p53 mutations in exons 5-9. Automatic sequencing was used to characterize the mutations and their effect on the P53 protein was analyzed. Bivariate analysis was used to assess the association with other prognostic factors. RESULTS: PCR-SSCP found no mutations in any control group patient, whereas 38.3% of patients with superficial transitional cell carcinoma had one or more mutations in the exons analyzed. Thirty mutations were sequenced; all were point mutations and 86.67% were considered relevant for the P53 structure. A total of 93.3% of the mutations were located in highly conserved regions and 73.3% in mutational hot spots. The highest cell differentiation grades and pT1 stage were associated with a higher incidence of p53 gene mutations. Previous recurrences and other tumor-related histological variables were not associated with a higher percentage of mutations. CONCLUSION: Mutations at p53 did not appear in healthy bladder mucosa and were significantly more frequent in pT1 and high-grade (G-II and G-III) tumors. All mutations detected were point mutations and most caused considerable P53 structural abnormalities, implying major repercussions on P53 function. These data suggest that certain p53 mutations may have prognostic value, even though they were not associated with other classic recurrence and tumor progression parameters. Future analyses of the progress of patients with superficial bladder transitional cell carcinoma and mutated p53 will help clarify this aspect. PMID- 15539840 TI - Relation between serum anti-p53 antibodies and microvessel density in bladder cancer patients. AB - INTRODUCTION: Etiology of serum anti-p53 antibodies in bladder cancer patients is still unknown. In this study we evaluated the relationship between serum anti-p53 antibodies and microvessel density in bladder cancer patients. MATERIALS AND METHODS: Seventy-six patients with transitional cell carcinoma of the urinary bladder were assessed prospectively (18 Ta, 30 T1, 28 T2>or =). Serum anti-p53 antibodies were detected by enzyme-linked immunosorbent assay. Tumor p53 overexpression was assessed by immunohistochemical staining. Vessels were stained immunohistochemically using an antibody against platelet endothelial cell adhesion molecule CD31. Spearman correlation test and t test were used for statistical analysis. RESULTS: Serum anti-p53 antibodies were positive in 25 (60%) of 41 tumor p53-positive patients. While the mean (SD, range) microvessel density was found to be 43 (7.59, 8-99) in patients who had positive serum anti p53 antibodies, it was found to be 23 (4.53, 6-98) in patients who had negative serum anti-p53 antibodies. There was a good correlation between serum anti-p53 antibodies and microvessel density (p<0.05). No correlation was found between tumor p53 expression and microvessel density (p>0.05). CONCLUSIONS: We found that there is a significant correlation between the microvessel density and serum anti p53 antibodies. This result may show the role of angiogenesis in the etiology of serum anti-p53 antibodies in bladder cancer patients. PMID- 15539841 TI - Comparative study of apoptotic antitumor effect between angiogenesis inhibitor AGM-1470 and MVAC chemotherapy on rat urinary bladder cancer. AB - OBJECTIVES: To investigate the antitumor apoptotic effect of AGM-1470 by comparing it to that induced by methotrexate, vinblastine, doxorubicin, and cisplatin (MVAC) therapy, currently the standard chemotherapy for bladder cancer, in a rat bladder cancer model. MATERIALS AND METHODS: A total of 45 six-week-old female rats were divided into 3 equal groups: those receiving AGM-1470 treatment; those receiving MVAC treatment, and controls. All rats were cystectomized to evaluate the therapeutic effect with regard to macroscopic tumor findings, hematoxylin and eosin pathology, apoptosis detection, and immunohistochemistry (IHC) for bcl-2. RESULTS: Our experimental protocol succeeded in producing invasive bladder tumors in the majority of rats. Tumor volume was significantly reduced in the AGM-1470 and MVAC treatment groups compared with that in the control group. The apoptotic indices of tumor cells was significantly higher in the AGM-1470 and MVAC treatment groups than in the control group. IHC for bcl-2 demonstrated no statistical difference in expression among the groups. However, the apoptotic indices of high-level bcl-2 expression were significantly lower than the indices of low-level expression in the AGM-1470 group. CONCLUSIONS: AGM 1470 and MVAC appear to exert a prominent mass reduction effect via tumor cell apoptosis in cases of invasive bladder tumor, although these therapies did not demonstrate any obvious modulation of bcl-2 protein expression status. Bcl-2 overexpression might be an obstacle to AGM-1470 therapy because of its significant inhibitory effect on apoptosis. PMID- 15539842 TI - Retroperitoneoscopic ureterolithotomy for the treatment of ureteral calculi. AB - INTRODUCTION: Most upper or middle ureteral stones are treated with shock wave lithotripsy or endoscopic techniques. In rare cases the ureteral stones are treated with open surgery after failure of first-line treatments. Retroperitoneoscopy is a minimally invasive alternative to open surgery. PATIENTS AND METHODS: Between May 1995 and January 2001, twenty-one patients underwent retroperitoneoscopic ureterolithotomy. The stones in upper and middle ureter were large and impacted (5 patients) or not fragmented after shock wave lithotripsy (16 patients). A balloon dissector was placed and infiltrated with 800 ml air in the retroperitoneal space. Three 10-mm trocars were used. The pressure was kept at 15 mm Hg by carbon dioxide insufflation. The stones were extracted from the ureter using a laparoscopic stylet. RESULTS: The stones in 17 patients were successfully removed in a median operating time of 105 (min-max 45-190) min. Urine extravasation in all cases and pnomoscrotum in 2 cases were observed as postoperative complications. The median hospital stay was 6 (min-max 3-22) days with minimal analgesic requirement. CONCLUSION: Retroperitoneoscopic ureterolithotomy is a useful and effective alternative treatment technique to open surgery when first-line treatments have failed or are unlikely to be effective. PMID- 15539843 TI - Extracorporeal shockwave lithotripsy compared with ureteroscopy for the removal of small distal ureteral stones. AB - INTRODUCTION: The treatment of small distal ureteral stones smaller or equal to 5 mm in size is still highly controversial. In distal ureteral stones larger than 5 mm in size, ureteroscopy (URS) has been shown in many studies to be superior to shockwave lithotripsy (SWL). The objective was to analyze the stone-free rate after treatment of distal ureteral stones with in situ SWL or URS. MATERIALS AND METHODS: A total of 3,857 SWL treatments were performed at our institution between 1996 and 2001. During this period 45 in situ SWL procedures were performed with the Dornier MFL 5000 lithotripter on distal ureteral stones regardless of the stone size. A total of 262 URS treatments were performed on distal ureteral stones. URS for small (5 mm or less) distal ureteral stones was performed in 110 cases. RESULTS: Distal ureteral stones smaller or equal to 5 mm in size were treated successfully stone free in 78% in one SWL session. Patients required a second SWL in 14% of the cases and 8% of the patients required a third SWL session. URS patients were successfully stone free after the procedure in 97% of the cases. Failed URS that needed an additional URS were performed in 2 and 1% of the patients had one SWL in situ treatment. CONCLUSIONS: URS treatment has shown to be the therapy of choice for distal ureteral stones. It is more effective than SWL treatment in this stone location. In experienced hands URS is a safe though even more invasive procedure than SWL. This can be expected as urologists perform more than 40 URS procedures per year. PMID- 15539844 TI - Hemostasis in tubeless PNL: point of technique. AB - INTRODUCTION: Tubeless PNL (percutaneous nephrolithotomy), in a highly selected group of patients, is a modification aimed at reducing the morbidity of PNL. We present a simple technique of achieving tract hemostasis as an adjunct to the safe performance of a tubeless PNL. MATERIALS AND METHODS: Charts of 40 consecutive patients who underwent tubeless PNL at our center were reviewed. In the latter 20 consecutive patients, diathermy coagulation of the intrarenal bleeders and tract was done and these patients were compared with the earlier 20 patients in whom fulguration was not done. Drop in hemoglobin, postoperative analgesic requirement, operating time, length of hospitalization and postoperative complications were compared. RESULTS: No statistical difference was found between the operative times and drop in hemoglobin for both the groups. The length of hospitalization and postoperative analgesic requirement were significantly less in the fulguration group. No significant complications were noted in either of the two groups. CONCLUSIONS: Fulguration of visible intrarenal and tract bleeders is a simple, safe and effective hemostatic adjunct in patients undergoing tubeless PNL. PMID- 15539845 TI - The smooth muscle ratio at the renal pelvis in adults: does it predict surgical outcome? AB - INTRODUCTION: Patients with ureteropelvic junction obstruction occasionally remain undiagnosed until adulthood. There are no objective criteria to predict the results of pyeloplasty in adult patients. We have evaluated the results of pyeloplasty in adult patients and investigated whether these results are correlated with the histopathology of the surgical specimen. MATERIALS AND METHODS: Histological sections from 26 patients with ureteropelvic junction obstruction were analyzed. Their mean age was 25.1 years. Diethylenetriamine pentaacetic acid scans were used to determine the degree of renal obstruction and postoperative healing. Improvement in diuretic renography was defined as a greater than 20% decrease in the half-time of the preoperative value. To examine smooth muscle and collagen tissue, sections were stained using Masson's trichrome. Smooth muscle ratio was identified by color image analysis. RESULTS: 18 patients (69.2%) fulfilled the criteria of healing. Patients with an improved scan had a mean smooth muscle percent (SMP) of 1.85+/-0.87 while subjects with no significant change in their diuretic scans had a mean SMP of 0.36+/-0.03 (p=0.001). There was a strong correlation between the SMP and the improvement. CONCLUSIONS: Adult pyeloplasty was found successful in about 70% of the cases. The SMP of the renal pelvis seems to be helpful in predicting the surgical outcome. PMID- 15539846 TI - Change in parameters before and after alpha-1-blocker therapy for men with lower urinary tract symptoms using color doppler ultrasound urodynamics: possible application for prediction of clinical outcome. AB - INTRODUCTION: We previously developed a noninvasive video urodynamic study using color Doppler ultrasonography. We sought the best flow velocity-related parameter which would allow prediction of an improvement in lower urinary tract symptoms (LUTS) after alpha 1-blocker treatment. METHODS: Twenty-two men with benign prostatic hyperplasia who were treated with a nonselective alpha 1-blocker (urapidil) were included. Subjective symptoms were evaluated using the International Prostate Symptom Score (IPSS) before and after alpha 1-blocker treatment. We measured the flow velocities using a transperineal ultrasound technique in the distal prostatic urethra just proximal to the external urethral sphincter (V1) and in the sphincteric urethra (V2), and used them to obtain the velocity ratio (VR=V1/V2). The corresponding functional cross-sectional areas of the urethra at these two sites (A1 and A2) were calculated as Q(max)/V. All these parameters obtained by the velocity-flow urodynamics were compared before treatment and after 4 weeks. RESULTS: After treatment, V1 and VR were decreased, and A1 was increased. V2 correlated best with the change in IPSS before and after alpha 1-blocker therapy, with Spearman's rho of 0.584. All men with V2 exceeding 50 cm/s did not show an improvement in the LUTS. CONCLUSIONS: The maximum flow velocity at the sphincteric urethra (V2) can predict the subjective outcome of alpha 1-blocker treatment. The velocity-flow parameters changed after alpha 1 blocker treatment. We confirmed that the transperineal ultrasound urodynamic study is not only noninvasive but also informative. PMID- 15539847 TI - Gyrus plasmasect: is it better than monopolar transurethral resection of prostate? AB - INTRODUCTION: This randomized prospective study was conducted to compare the efficacy and safety of the Gyrus Plasmasect loop bipolar transurethral resection of prostate (TURP) and conventional monopolar TURP in the treatment of benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 117 men were enrolled in this study. Fifty-eight patients underwent Gyrus Plasmasect TURP and 59 patients underwent monopolar TURP. They were followed up for 3 months after surgery. RESULTS: Significant improvements were seen postoperatively in both the Gyrus and monopolar groups in terms of prostatic volume, International Prostate Symptom Score, quality of life score, peak flow rate, and post-void residual urine volume. However, the degree of improvement was not statistically different between the 2 groups. Significantly less blood loss, shorter postoperative catheterization time and length of hospital stay were seen in the Gyrus group. CONCLUSIONS: Gyrus Plasmasect TURP yielded comparable results to monopolar TURP; however, this is only a preliminary study and follow-up is necessary to assess its long-term efficacy. PMID- 15539848 TI - Bladder neck stenosis after transurethral resection of prostate: does size matter? AB - AIM: To understand the risk factors associated with the incidence of bladder neck stenosis (BNS) after transurethral prostate surgery. PATIENTS AND METHODS: We retrospectively reviewed 900 patients who underwent transurethral prostate surgery over a 4-year period. The mean age of the men was 72.3 (47-94) years. The specific outcome data assessed related to BNS, including type of operation performed, resected tissue weight and history of previous surgery in the lower urinary tract. RESULTS: 29 (3.4%) patients developed BNS at a mean of 10.3 (3-33) months, with a mean resected prostatic tissue weight of 11+/-3.7 g. Four of the 29 patients with BNS were treated with bladder neck resection and re-stenosed. Fifty-four men underwent bladder neck incision for small prostates with a high bladder neck, measured by digital rectal examination and assessed cystoscopically, with no BNS. All the remaining patients from our series did not have a BNS, with a mean resected weight of 28+/-8.9 g, which is statistically greater than in the BNS group (p<0.05, unpaired t test). CONCLUSIONS: BNS after transurethral prostate surgery is a significant problem. It is clear from our study that resection in small prostates with no sign of a high bladder neck will increase the development of BNS. Thus, small prostates should be managed by an initial bladder neck incision, even if the bladder neck is not high. PMID- 15539849 TI - Transurethral prostatectomy and inguinal hernia repair in a single session. AB - INTRODUCTION: We aimed at evaluating the outcomes of transurethral prostatectomy and inguinal hernia repair performed in a single session. PATIENTS AND METHODS: Fifty-six patients (mean age 68+/-8.3 years) in whom transurethral prostatectomy was performed combined with an inguinal hernia repair were included into the study. Type of anesthesia, technique of inguinal hernia repair, hospitalization time, and the complications encountered were recorded. Cost comparisons were made using the official price-lists of the Turkish Medical Association. All patients were asked whether they were satisfied with the outcome of both operations performed in one session. The data obtained from the patients who underwent both operations in one session were compared with those obtained from 56 patients who underwent transurethral prostatectomy only (control group). Statistical analysis was performed using the chi-square test corrected for continuity according to the Yates or the Fisher exact test. RESULTS: The operations were performed in 19 patients under general, in 20 patients under epidural, and in 14 patients under spinal anesthesia. Three patients were given general anesthesia and spinal anesthesia combined. In 6 patients bilateral and in 50 patients unilateral hernia repair was performed. In 11 repairs, polyprolene mesh grafts were utilized; in 2 repairs, a laparoscopic method was used, and in the remaining 49 repairs, one of the conventional techniques (McVay, Bassini, or Shouldice) was employed. There were no significant differences with regard to early and late postoperative complications and satisfaction between study group and control group (p>0.05). Combined prostatectomy and hernia repair allows approximately 30% cost profit. CONCLUSIONS: Performing transurethral prostatectomy and inguinal hernia repair in one session decreased the number of the operations and anesthesias, hospital stay, and thus health costs and did not cause an increase in operative and postoperative morbidity. PMID- 15539850 TI - Paraganglioma of the bladder: controversy regarding treatment. AB - Paragangliomas or extra-adrenal pheochromocytomas account for approximately 15% of all pheochromocytomas. They are usually found in an intradiaphragmatic location. Paragangliomas of the urinary bladder are infrequent and represent 6% of the paragangliomas and 0.06% of all bladder tumors. We describe 2 cases of urinary bladder paragangliomas attended to our center and review the literature with special attention to treatment, since there is some controversy around the management of these tumors. Surgical options include partial or total cystectomy and transurethral resection, but there is no consensus regarding these treatment modalities. The 2 patients presented underwent transurethral resection with a good outcome and no recurrence or residual disease. PMID- 15539851 TI - Development of leiomyosarcoma from primary leiomyoma? AB - Kidney leiomyosarcoma represents a rare variety of malignant kidney tumours. In this paper, we report on a patient with an inoperable leiomyosarcoma. Since this neoplasm is very rare, there is very little information on this type of malignancy. We present the symptoms, radiological findings, diagnostic criteria and differential diagnosis of the tumour. Leiomyosarcoma exhibits aggressive biological behaviour and has a poor prognosis. We have found that the treatment of choice is a radical nephrectomy. PMID- 15539852 TI - Localized primary amyloidosis of the genitourinary tract: does conservatism help? AB - Primary amyloidosis of the genitourinary tract is a rare clinical entity with an excellent prognosis in most cases. It is of interest to the urologist as it mimics malignancy. The definitive diagnosis is made on histologic examination and by excluding secondary causes of amyloidosis. We present 4 patients with primary localized amyloidosis of the genitourinary tract who were treated with transurethral resections alone with satisfactory outcome. PMID- 15539853 TI - Bilateral renal artery stenting in a patient with Leriche syndrome. AB - Stenting for renal artery stenosis is well described in the literature. Bilateral renal artery stenting is not such a common procedure, however it is quite rare in patients with Leriche syndrome, as is the case we present. PMID- 15539854 TI - Diplopia - an unusual primary manifestation of metastatic renal cell carcinoma. AB - It is relatively unusual that initial symptoms of renal cell cancer begin with the metastatic involvement of other sites. Intracranial metastases especially in the paranasal sinuses may be unusual not only to surgeons but also to physicians. In this report, we present a case where a metastasis was first manifest as a sphenoid sinus secondary with ocular and visual disturbances prior to the demonstration of the primary tumor site. It turned out to be a case of diplopia masquerading as a metastatic renal cell cancer. PMID- 15539855 TI - Vesicoureteral reflux after intravesical instillation of bacillus Calmette-Guerin against carcinoma in situ of the bladder. AB - We report a case of vesicoureteral reflux (VUR) 3 years after intravesical instillation of bacillus Calmette-Guerin (BCG, Tokyo 172 strain) against carcinoma in situ of the bladder. The present case suggests that a long-term careful follow-up is needed to detect not only tumor recurrences but also VUR as a late complication after intravesical BCG instillation. PMID- 15539856 TI - Noradrenergic mechanisms in the pathophysiology of post-traumatic stress disorder. AB - Post-traumatic stress disorder (PTSD) is a serious psychiatric illness that may develop in individuals after exposure to a traumatic event. Recent data suggest that trauma and/or long-term stressors can cause alterations in the functioning of neuroanatomical structures and neural networks throughout the central nervous system. Specifically, dysregulation in central and perhaps, peripheral noradrenergic neural networks has been implicated as the cause of specific symptom clusters in the pathophysiology of PTSD. In this review, both clinical and preclinical data are presented to highlight types of noradrenergic dysfunction observed in individuals with PTSD. Additionally, the role of noradrenaline dysregulation in the acquisition/initiation, and maintenance of hyperarousal and reexperiencing symptom clusters in PTSD will be addressed. PMID- 15539857 TI - Possible association between serotonin transporter promoter region polymorphism and extremely violent crime in Chinese males. AB - The neurotransmitter, serotonin, has been implicated in aggressive behavior. The serotonin transporter (5-HTT), which reuptakes serotonin into the nerve terminal, plays a critical role in the regulation of serotonergic function. Previous western reports have demonstrated that the low-activity short (S) allele of the 5 HTT gene-linked polymorphic-region (5-HTTLPR) polymorphism is associated with aggressive behavior and associated personality traits. In the present study, we investigated this 5-HTTLPR genetic polymorphism in a group of Chinese males who had been convicted for extremely violent crime (n = 135) and a normal control group (n = 111). The proportion of S-allele carriers was significantly higher in the criminal group than in the controls (p = 0.006). A significant association was not demonstrated for the relationship between the 5-HTTLPR polymorphism and antisocial personality disorder, substance abuse or alcohol abuse in the criminal group. Our findings demonstrate that carriage of the low-activity S allele is associated with extremely violent criminal behavior in Chinese males, and suggests that the 5-HTT may be implicated in the mechanisms underlying violent behaviors. PMID- 15539858 TI - Association analysis for MAOA gene polymorphism with long-latency auditory evoked potentials in healthy females. AB - The components of auditory evoked potentials (AEPs) have been demonstrated to be heritable and to be influenced by different neurotransmitter activities such as serotonin or dopamine. Monoamine oxidase A (MAOA) is a mitochondrial enzyme involved in the degradation of dopamine, serotonin and norepinephrine. We examined the potential association of the functional 30-bp repeat MAOA gene linked polymorphism region (MAOA-LPR) in 234 normal young Chinese females. The results demonstrated that there is no association for MAOA-LPR polymorphism and AEP components. Our negative findings suggested that this genetic polymorphism does not play a major role in the modulation of AEPs in normal young females, but we could not exclude that other polymorphisms of the MAOA gene affect AEP components. Further exploration of the other polymorphisms of the MAOA gene and multiple interactions of the polymorphisms in various neurotransmitter systems may be needed in future studies. PMID- 15539859 TI - Association analysis for the C-1019G promoter variant of the 5-HT1A receptor gene with auditory evoked potentials in major depression. AB - Involvement of the serotonergic system in N1 and P2 components of auditory evoked potentials (AEPs) has been implicated. Moreover, studies have indicated the presence of heritability in the genesis of AEP components. The serotonin 1A (5 HT1A) receptor gene is a strong candidate for N1 and P2 components of the AEPs because 5-HT1A receptor regulates the firing of serotonergic neurons. The present study tested the hypothesis that the 5-HT1A promoter genetic polymorphism (C 1019G) is associated with N1 and P2 components of AEPs in unmedicated major depression patients. The sample consisted of 221 Chinese patients (mean age: 44.3 years; male/female: 93/128) diagnosed with major depression. AEPs and 5-HT1A genotyping were done for each patient. Patients with the C/C genotype had a significantly shorter P2 latency when compared with C/G or G/G genotype patients (p = 0.049), and the difference in P2 latency was significant among the 5-HT1A genotype groups in male patients (p = 0.031) but not in female patients (p = 0.398). These findings suggest that this 5-HT1A polymorphism may affect AEP P2 latency in a gender-dependent manner. Further studies with other genetic polymorphisms in the serotonergic system may help to clarify the relation between serotonergic function and AEP components. PMID- 15539860 TI - Reduced white matter volume of the caudate nucleus in patients with schizophrenia. AB - To examine volumetric abnormality, the caudate nucleus was measured in patients with schizophrenia and healthy subjects using magnetic resonance imaging. The absolute and relative volumes of white matter in the caudate nucleus were found to be significantly smaller in patients with schizophrenia compared to those in healthy subjects. There were significant correlations between dosages of neuroleptics during the previous year and absolute gray matter volumes of the caudate nucleus as well as relative white matter volumes in patients with schizophrenia. These findings suggest that reduced white matter volume of the caudate nucleus may be a factor affecting the abnormal connectivity of the corticostriatal loop, and that neuroleptic medication would be related to white matter alteration in patients with schizophrenia. Our result replicates a previous study reporting that there is a lack of negative correlation between age and caudate nucleus volume. We also suggest that the disease process of schizophrenia might interfere with normal aging. PMID- 15539861 TI - Positive association between T-182C polymorphism in the norepinephrine transporter gene and susceptibility to major depressive disorder in a japanese population. AB - Norepinephrinergic neurotransmission in the central nervous system appears to have a major impact on the symptomatology in major depressive disorder and the human norepinephrine transporter (NET) gene is one of the key candidates for genetic studies in major depressive disorder. The authors established a new allele-specific PCR-based genotyping procedure and examined whether the NET T 182C polymorphism was associated with the susceptibility to major depressive disorder in a Japanese population. This study included 145 patients with major depressive disorder (according to DSM-IV) and 164 healthy volunteers. There was a significant difference in the genotype distribution between major depressive disorder patients and healthy volunteers (p = 0.02), and the C/C genotype was associated with lesser susceptibility to major depressive disorder. The NET T 182C polymorphism may be in part related to the pathophysiology of major depressive disorder in a Japanese population. PMID- 15539862 TI - Decreased levels of dopamine D3 receptor mRNA in schizophrenic and bipolar patients. AB - Previous studies found an elevation of the dopamine D3 receptor (DRD3) mRNA as determined in peripheral lymphocytes in schizophrenic patients. The aim of this study was to test the hypothesis of elevated DRD3 mRNA in schizophrenia compared to bipolar disorder. Twenty-four patients, 13 schizophrenic and 11 bipolar, were included according to DSM-IV criteria. Psychometric measures were conducted using the Scale for the Assessment of Positive Symptoms, Scale for the Assessment of Negative Symptoms, Brief Psychiatric Rating Scale, Montgomery-Asberg Depression Rating Scale and Young Mania Rating Scale. mRNA was isolated from lymphocytes of venous blood samples and DRD3 mRNA was quantified using real-time reverse transcription PCR. We found a decrease in DRD3 mRNA in 13 schizophrenic (p = 0.009) and 11 bipolar (p = 0.023) patients as compared to controls. Medication history and severity of positive symptoms did not significantly influence DRD3 expression. Higher levels of DRD3 mRNA were correlated with negative schizophrenic symptoms. Interestingly, after treatment of patients with antipsychotics, DRD3 mRNA levels increased to similar levels as those of healthy controls. Bipolar patients, however, showed a slower increase in DRD3 mRNA levels after 3 weeks of therapy. Our findings suggest that the expression of DRD3 mRNA is reduced in schizophrenia and bipolar disorder, supporting the hypothesis of distorted homeostasis of dopamine receptor subtypes in psychotic disorder. The observed diminution was not specific for schizophrenia but also for bipolar disorder requiring further analysis of the regulatory factors involved in dopamine receptor subtype expression. PMID- 15539864 TI - Evaluation of the prophylactic efficacy of amitriptyline and citalopram, alone or in combination, in patients with comorbidity of depression, migraine, and tension type headache. AB - Antidepressants are used to treat chronic daily headache disorders such as migraine and chronic tension-type headache (TTH), which are often associated with depression and anxiety. Here, we studied the efficacy and tolerability of amitriptyline and citalopram, given alone or in combination, in patients with 'triple' comorbidity of depression, TTH, and migraine. Eighty-eight patients were enrolled in the study and randomly divided into two groups. The first group received amitriptyline and the second citalopram for 16 weeks. Patients were assessed at weeks 0, 4, 8, and 16. The two drugs were equally efficacious in relieving depressive symptoms, although amitriptyline was more efficacious than citalopram in reducing migraine and TTH attacks. Patients who did not respond to monotherapy (<30% of improvement in the clinical scores) were treated with a combination of the two drugs for 16 additional weeks. In these selected patients, the combined treatment produced a substantial improvement in depression, migraine and TTH without producing major side effects such as those commonly related to the 'serotonergic' syndrome. The results indicate that a combined therapy with amitriptyline and citalopram may be particularly beneficial for patients with TTH, migraine and comorbid depression that do not respond to monotherapy. PMID- 15539863 TI - Central and peripheral evaluation of rupatadine, a new antihistamine/platelet activating factor antagonist, at different doses in healthy volunteers. AB - AIMS: To assess peripheral anti-H1 and central nervous system (CNS) activity of single increasing doses of rupatidine fumarate (RU), a new antihistamine/platelet activating factor antagonist compound, in comparison with hydroxyzine and placebo. METHODS: Eighteen healthy young subjects of both sexes took part in a crossover, randomised, double-blind, placebo-controlled study. Treatments tested were: RU 10, 20, 40 and 80 mg and hydroxyzine 25 mg, as a positive standard. Before and several times after drug intake, peripheral anti-H1 activity was appraised by the skin reactivity to intradermal injection of histamine. CNS effects were also obtained by objective tests of psychomotor performance and subjective mood scales. RESULTS: All active treatments showed a significant reduction of the wheal and flare reaction in relation to placebo, RU displaying a potent dose-dependent inhibition pattern. The global nonparametric Friedman test to changes from placebo in 15 objective variables from psychomotor performance showed a significant impairment of similar magnitude after hydroxyzine 25 mg (p = 0.01) and RU 80 mg (p = 0.02), but this was slower in development and recovery after the latter. After RU 40 mg, a smaller impairment was also obtained (p = 0.04). Activity (p = 0.01) and drowsiness (p = 0.02) scales showed significant changes, the subjects feeling less active and more drowsy after all active treatments. CONCLUSION: RU presents a potent dose-dependent peripheral anti-H1 activity, displaying psychomotor impairment activity only at the highest dose (80 mg), while therapeutically relevant lower doses (10 and 20 mg) were similar to placebo. PMID- 15539865 TI - MDMA (3,4-Methylenedioxymethamphetamine) or ecstasy: the neuropsychobiological implications of taking it at dances and raves. AB - MDMA (3,4-methylenedioxymethamphetamine) or 'ecstasy' is a ring-substituted amphetamine derivative, which is widely used as a recreational drug, most particularly at dances and raves. Around 80-95% of dancers/ravers report using ecstasy/MDMA, compared to 5-15% of young people in general. This paper will consider the possible contribution of stimulatory environmental conditions to the neuropsychobiological effects of MDMA. Animal research shows that heat and crowding potentiate the acute effects of MDMA. Social interaction and intravenous drug self-administration in laboratory rats are significantly enhanced when MDMA is given under hot ambient temperatures. Loud noise and physical activity can also contribute to the general overarousal. Furthermore, MDMA impairs homeostatic thermal control in rats, leading them to overheat in hot environments. The human implications of these findings are that the hot, noisy and overcrowded conditions at raves may be providing the ideal environment to heighten the acute drug response. In recreational users, the acute medical dangers of MDMA comprise a constellation of hyperthermia-related abreactions, which generally only occur when it has been taken in hot and crowded environments. MDMA is well established as a serotonergic neurotoxin in laboratory animals, but heat and overcrowding increase the degree of distal axon terminal loss. If this also occurs in humans, then the stimulatory environments of clubs and raves may heighten the likelihood of adverse neuropsychological sequelae in recreational ecstasy users. Consistent with this prediction, the extent of self-reported dancing/exercise when on MDMA has recently been shown to be associated with significantly more psychobiological problems afterwards. PMID- 15539866 TI - Involvement of the noradrenergic system in performance on a continuous task requiring effortful attention. AB - To determine the effects of noradrenergic neuronal activity on performance in continuous tasks requiring effortful attention, the performance of 23 male students in the Uchida-Kraepelin test (UKT) was examined. The UKT requires continuous arithmetic addition of single-digit figures for 25 min. The relationship of performance with saliva levels of 3-methoxy-4-hydroxyphenylglycol (sMHPG) was analyzed. Saliva samples were taken before, during and after test performance, and sMHPG levels determined by gas chromatography-mass spectrometry. There was no significant change in mean sMHPG as a result of test performance. However, when initial effort was calculated, defined as number of items completed during the 1st min subtracted from the average completed per minute in the 1st and the 2nd halves (blocks) of the test, significant correlations with sMHPG (p = 0.0002 for the 1st block and p < 0.0001 for the 2nd block) were found. Thus the data indicate that noradrenergic neuronal activity affects the performance on continuous tasks requiring effortful attention. PMID- 15539867 TI - Prenatally detected cystic adrenal mass associated with Beckwith-Wiedemann syndrome. AB - We report a case of a right-sided cystic adrenal mass, detected after the 21st week of gestation, associated with fetal macrosomia. The diagnosis of Beckwith Wiedemann syndrome was evoked. Prenatal sonography and magnetic resonance imaging did not allow establishing the origin of the suprarenal mass. The differential diagnosis of cystic neuroblastoma, pseudocystic adrenal haemorrhage, and adrenocortical macrocysts was discussed. A laparotomy was performed 2 weeks after birth due to the increasing size of the tumour and due to its possible malignant origin. Histological findings were haemorrhagic lesions without evidence of malignancy and adrenal cortical cytomegaly. The diagnosis of an adrenocortical macrocyst component of Beckwith-Wiedemann syndrome was established. PMID- 15539868 TI - Effective counselling of pre-natal diagnosis of serious heart disease--an aid to maternal bonding? AB - The pre-natal diagnosis of a serious anomaly in the fetus precipitates a crisis for the parents. Effective counselling may help during this difficult phase. We postulated that if parents opted to continue with the pregnancy, then they are better prepared to care for the newborn infant having already experienced the grieving process. OBJECTIVES: This paper reviews our early experience, focusing on the parental perception of the counselling. METHODS AND SUBJECTS: Questionnaires were developed and forwarded to patients previously seen in a private obstetric ultrasound practice, diagnosed with or specifically referred because of the diagnosis of a complex cardiac abnormality in the fetus. The diagnosis was subsequently confirmed by a further scan in the presence of a paediatric cardiologist, who initiated explanation of the anomaly with the obstetrician ultrasonologist asking and seeking explanations in lay terms on behalf of the parents, concerning the nature of the anomaly, its possible consequences, the need for intervention, the short- and long-term risks, concentrating on quality of life issues. RESULTS: Over a 3-year period a cohort of 40 such patients were reviewed of whom a third opted for termination. The anomalies were complex and many life-threatening, most requiring surgical intervention in the newborn period or subsequently. Of the 27 that proceeded to term, responses were obtained from 21. Most described the shock at learning the diagnosis and of being very distressed at the time. Most commented that the information provided was 'just right'. The majority felt that the previous contact with the obstetrician and cardiologist was very helpful in their subsequent care of their newborn infant. Three parents noted that they continued to experience anxiety about their baby, while 3 others indicated that the previous counselling had 'no effect' on the handling of their baby. Anecdotal observations made of the parents especially of the mother's handling of the newborn infant seemed to support the parents' perception that they were helped by their prior knowledge of the diagnosis. CONCLUSION: With increasing pre-natal diagnosis of major anomalies, it is important that parents so affected have the opportunity to understand the nature of the abnormality, its significance and effects on the infant's and child's functioning and survival, and the need for intervention. Such information together with working through the grieving process during the latter part of the pregnancy may help with the handling of the infant once born and further improve their bonding with their baby, despite the need for, at times, life-threatening intervention. Further objective studies are required to confirm these preliminary observations. PMID- 15539869 TI - Second- and third-trimester therapeutic terminations of pregnancy in cases with complete placenta previa--does feticide decrease postdelivery maternal hemorrhage? AB - OBJECTIVE: To study the feasibility of second- and third-trimester termination of pregnancy (TOP) with complete placenta previa, and the impact of performing feticide before labor induction on maternal hemorrhagic morbidity. PATIENTS AND METHODS: From 1987 to 2002, the databases of two referral hospitals were reviewed. We identified 15 cases of second- or third-trimester TOP in women with complete placenta previa. Feticide was performed 2-14 days before induction in 6/15 cases. Cervical ripening was achieved in 8 cases by mifepristone alone (n = 2) or by mifepristone and dilapan (n = 6). Labor was induced by vaginal gemeprost (n = 2), intramuscular (n = 5) or intravenous (n = 4) sulprostone, vaginal misoprostol (n = 1) or a combination of misoprostol and sulprostone (n = 3). Hemorrhage was defined by the need for transfusion. The difference between the preoperative and the lowest per- or postoperative maternal hemoglobin level was also analyzed. RESULTS: Of the 9 women who underwent labor induction without previous feticide, 4 required blood transfusions, 1 of whom had a hemostat hysterectomy. The mean hemoglobin difference was 2.5 g/dl (range: 0.5-5.3). None of the 6 patients with preinduction feticide required transfusion. The hemoglobin difference was significantly smaller in this group than in terminations without previous feticide (mean: 1.0 g/dl ; range: 0.1-2.2; p = 0.03). CONCLUSION: In cases with complete placenta previa, second- or third-trimester TOP is feasible. It carries a substantial risk of hemorrhage that might be decreased by preinduction feticide. PMID- 15539870 TI - Sonographic, pathologic and karyotypic findings in a rare case of placenta fenestrata. AB - Abnormalities of placental shape are occasionally seen on ultrasound. They have not been reported to be associated with abnormalities in fetal anatomy and karyotype. Here, we report on a rare case of placenta fenestrata with triploid karyotype. A 15-year-old patient presented at 21 weeks and 3 days gestation for ultrasound evaluation following an abnormal triple screen and abnormal ultrasound. Multiple fetal abnormalities were noted as well as several cystic areas with pulsatile flow on Doppler ultrasound in the placenta. After termination of the pregnancy, a rare abnormality in the placental shape, placenta fenestrata, was noted. The fetal karyotyping showed a triploid karyotype. This is the first reported case of placenta fenestrata diagnosed on ultrasound. In addition, this is also the first reported case of karyotype abnormality associated with abnormality of placental shape. PMID- 15539871 TI - Elevated second-trimester free beta-hCG as an isolated finding and pregnancy outcomes. AB - OBJECTIVE: To investigate the relationship between unexplained elevated second trimester free beta-human chorionic gonadotropin (beta-hCG) levels and pregnancy complications as well as adverse pregnancy outcomes. METHODS: The study cohort comprised 2,110 non-smoking women with chromosomal and structurally normal fetuses at low-risk for both Down's syndrome (risk <1:250) and neural tube defects (maternal serum alpha-fetoprotein <2.0 MoM). A free beta-hCG value of > or =2.0 MoM was used to define the populations with elevated levels of free beta hCG. Descriptive statistics, chi2 test, Fisher's exact test, and logistic regression analysis were used for statistical analysis, and p < 0.05 was considered statistically significant. RESULTS: The mean maternal age of the study group was significantly lower than in controls (27.9 +/- 4.3 and 30.6 +/- 5.1 years, respectively, p < 0.05), while the proportion of primigravidas was significantly higher compared to that of controls (p < 0.05). After adjustment of the 2 groups according to maternal age and parity, we observed an increased incidence of preeclampsia among women with elevated free beta-hCG levels in relation to controls (p < 0.05). However, a logistic regression analysis demonstrated that the free beta-hCG level was not a predictor of the occurrence of preeclampsia. No significant relationship was found with the incidence of gestational diabetes, oligohydramnios, polyhydramnios, pregnancy-related hypertension, intrauterine growth retardation, preterm delivery, spontaneous abortion and stillbirths (p > 0.05). PMID- 15539872 TI - Prenatal diagnosis of trisomy 2 mosaicism: a case report. AB - We report a case of trisomy 2 mosaicism detected upon amniocentesis in a woman with advanced maternal age. A mos 47,XY,+2(4)/46,XY(21) karyotype was revealed using standard GTG banding. There were no pathological sonographic findings and the fetal size was normal for gestational age at 16th week. The use of serial high-resolution ultrasound examination of the fetus to detect major abnormalities was offered as an option to the parents who, however, decided for termination of the pregnancy. Fetal autopsy did not reveal any malformations. Trisomy 2 mosaicism is associated with variable phenotypic abnormalities without a specific pattern, intrauterine growth restriction, fetal demise or stillbirth. The rarity of trisomy-2 mosaicism in prenatal diagnosis, as well as the increased risk of an abnormal outcome makes the diagnostic approach and genetic counseling difficult. PMID- 15539873 TI - Cephalic index is not a useful sonographic marker for trisomy 21 and trisomy 18. AB - OBJECTIVES: To evaluate whether the cephalic index (CI) in trisomy 21 (T21) and trisomy 18 (T18) fetuses is different from that in euploid fetuses, and if so, is this difference of clinical utility. METHODS: Retrospective. Over an 18-month period, patients attending a single centre for routine advanced maternal age amniocentesis were recruited for a prospective study of ultrasound soft markers of aneuploidy. This prospective database was searched for cases with the following criteria: (1) occipitofrontal diameters (OFD) measured at least twice; (2) gestational age between 98 and 126 days either by ultrasound-confirmed menstrual dates or early second- trimester biometry; (3) no major central nervous system abnormalities detected on prenatal ultrasound, and (4) normal fetal karyotype. This constituted the control group. The study group consisted of all cases of prenatally diagnosed T18 and T21 identified in the same time period with criterion 2 as above. The fetuses in the study group had the OFD measured in a blinded fashion from the biparietal diameter images. CI (= mean biparietal diameter/mean OFD) was calculated for all fetuses. Pearson coefficient and regression analysis were used to determine independence of CI to gestational age in the control group. Standard descriptive statistics were used to describe interval data and two-tailed t test was used to compare means between the study and control groups. ROC curves were constructed to evaluate the clinical efficacy of CI for T18 and T21. RESULTS: Five hundred and ninety-seven fetuses were available for analysis. There were 551 fetuses in the control group and 46 in the study group. Within the study group, there were 30 T21 and 16 T18 fetuses. Within the control group, CI was independent of gestational age (R = 0.026, p = 0.922). Mean CI for the control group was 0.802 (SD 0.040) and this was not statistical different from either the T21 group (mean 0.816, SD 0.042, p = 0.067) or the T18 group (mean 0.792, SD 0.057, p = 0.491). Area under the ROC curves was determined for both T18 and T21 and both had poor results (0.545 and 0.598, respectively). When CI was evaluated in the control group according to the two main ethnic groups in the study, there was a trend towards a statistical difference (p = 0.046) between the fetuses of Oriental and Caucasian mothers. CONCLUSIONS: In this retrospective study, CI was not found to be statistically different between the study and control groups. Although a trend towards significance was seen with T21, this difference is not clinically useful. There may be interethnic differences in the CI between fetuses. CI is not useful for aneuploidy screening by ultrasound. PMID- 15539874 TI - Prenatal diagnosis of common aneuploidies using multiplex quantitative fluorescent polymerase chain reaction. AB - OBJECTIVE: Prenatal diagnosis of foetal trisomies is usually performed by cytogenetic analysis. This requires lengthy laboratory procedures and it is expensive. Here, we report a retrospective study of quantitative fluorescent polymerase chain reaction (QF-PCR) for prenatal detection of trisomies 13, 18 and 21. METHODS: QF-PCR was performed on a total of 447 amniotic fluids blindly analysed without any knowledge of the cytogenetic results and 43 samples with known karyotype. All samples were tested with at least 4 small tandem repeat markers specific for each chromosome 13, 18 or 21. RESULTS: QF-PCR results on amniotic fluid were consistent with conventional cytogenetic data. QF-PCR detected 5 cases of trisomy 21, 2 cases of trisomy 18, 1 case of trisomy 13 and 1 case with Klinefelter's syndrome. CONCLUSIONS: QF-PCR has proved to be very useful in clinical settings, since it allows the detection of major numerical disorders in a few hours after sampling and thus reduces parental anxiety. PMID- 15539875 TI - Amniotic fluid and maternal blood characteristics in severe mid-trimester twin twin transfusion syndrome. AB - OBJECTIVE: To investigate the concentrations of metabolic variables in the amniotic fluid of the recipient twin and maternal blood and to correlate them with clinical features, which are characteristic for the course of pregnancies with twin-twin transfusion syndrome (TTS). MATERIALS AND METHODS: In 109 pregnancies with severe mid-trimester TTS, we measured the concentrations of interleukin-6 (IL-6), alpha-fetoprotein (AFP), sodium, potassium, total protein, beta2-microglobulin and osmolality in the amniotic fluid of the recipient twin prior to laser coagulation of the vascular anastomoses and compared them to a control group of 92 singleton pregnancies. We measured the pulsatility index (PI) of ductus venosus flow velocity waveforms in the recipient twin and performed a retrospective chart analysis for complete maternal blood count before and after the procedure. RESULTS: All variables, except osmolality, IL-6 and AFP were significantly lower in the TTS group. There was a significant positive correlation between PI in the ductus venosus and the amniotic fluid AFP concentrations (r = 0.355; p < 0.001). There was a significant fall in complete maternal blood count after laser therapy (p < 0.001) and a significant correlation between the amount of amniotic fluid drained and the fall of maternal Hb (r = -0.261; p = 0.03) and hematocrit (r = -0.212; p = 0.01). CONCLUSION: Pathophysiologic changes in TTS do not only occur at the inter-twin level of placental vascular anastomoses but also at the materno-fetal level of fluid exchanges. AFP is correlated to the severity of congestive heart failure of the recipient twin. PMID- 15539876 TI - Association of deletions of the chromosomal region 15q24-ter and diaphragmatic hernia: a new case and discussion of the literature. AB - OBJECTIVE: To provide new insights into how chromosomal aberrations affect fetal development, as well as for the counseling of parents in comparable situations, it is important to characterize and report the genotypes of fetuses with clinical anomalies. METHODS: Molecular cytogenetic analyses in a fetus with congenital diaphragmatic hernia (CDH). RESULTS: This report describes the first case of a deletion of the region q26.1-ter on chromosome 15 occurring as a de novo event associated with CDH. A detailed review of the literature provides further evidence of a functional association between deletions within the chromosomal region 15q24-ter and the development of CDH. CONCLUSIONS: The obtained data argue that detection of such a deletion in the region 15q24-ter associated with CDH likely predicts a poor prognosis. This report highlights the importance of giving special diagnostic attention to the chromosomal region 15q24-ter when prenatal ultrasound examination provides evidence of a CDH and warrants further research to identify genetic elements within the chromosomal region 15q24-ter related to the development of diaphragmatic hernia. PMID- 15539877 TI - The University of California at San Francisco Fetal Treatment Center: a personal perspective. AB - Today, the Fetal Treatment Center at the University of California, San Francisco is premier not only for its innovative and cutting-edge developments in fetal diagnosis and therapy, but in its delivery of excellent clinical care to patients from within and outside the United States. Indeed it is recognized as the pioneering institution for fetal surgery. But, the road to success and international recognition has been torturous and often tumultuous. From a personal perspective, Dr. Michael Harrison recounts the center's 25-year history- an evolution that has seen a 'California fault zone' become an epicenter of excellence for fetal therapy. Along the way, Dr. Harrison recalls the best and worst of times in the center's history particularly as they relate to fetal surgery, recalls the initial principal payers and subsequent cadre of professionals responsible for developing the center and providing the medical and surgical expertise that is the hallmark of the UCSF Fetal Treatment Center, provides a subjective assessment, a 'report card', of where they stand in fetal therapy today, and finally, gives well-deserved credit to the many talented research fellows, whom he calls the 'heart and soul' of the enterprise, whose 'fingerprints' are borne on all the advances in fetal treatment. PMID- 15539878 TI - Sonographic findings of fetuses with an empty renal fossa and normal amniotic fluid volume. AB - OBJECTIVE: To review the antenatal sonographic findings and postnatal follow-up of fetuses with empty renal fossa (ERF) and normal amniotic fluid volume. METHODS: Sonographic examinations of 13,705 fetuses were retrospectively analyzed and all fetuses with at least one ERF and normal antenatal amniotic fluid volume were included in this study. RESULTS: Forty cases with antenatal ERF were diagnosed. Prenatal diagnosis consisted of pelvic kidney (n = 24), unilateral renal agenesis (n = 13), horseshoe kidney (n = 2) and crossed fused renal ectopia (n = 1). The prevalence of ERF in the low-risk population approximated 3.2 per thousand. There was no serious renal complication during a mean follow-up period of 30 months except one case of crossed fused renal ectopia requiring hemodialysis. CONCLUSION: The underlying cause of ERF in the majority of cases with normal amniotic fluid volume is renal ectopia. Prenatal ultrasonography seems to be highly reliable in diagnosing these anomalies. Prognosis is favorable in the absence of additional extraurinary malformations. PMID- 15539879 TI - In utero meconium suctioning may prevent meconium aspiration. AB - OBJECTIVE: To assess usefulness of in utero meconium suctioning using endoscopic approach. MATERIALS AND METHODS: In utero meconium suctioning under direct endoscopic visualization was successfully performed in 11 fetuses in whom thick meconium was detected upon rupture of the membranes. A gas-sterilized flexible fiberoptic scope with an operational channel was introduced via cervix, and meconium was aspirated from the fetal mouth. Definitive suction was performed in all neonates immediately after birth. RESULTS: Endoscopic meconium suctioning was successfully performed in 11 out of 16 fetuses with retrieval of 3-12 ml of meconium. Only 1 neonate in this series of cases had meconium below the vocal cords and none developed meconium aspiration syndrome postnatally. CONCLUSION: Because fatal meconium aspiration syndrome is largely an intrauterine event, the concept of in utero prophylactic meconium suctioning may serve as an effective modality for its prevention. PMID- 15539880 TI - Hemodynamic effects of carvedilol infusion and the contribution of the sympathetic nervous system in rats with heart failure. AB - We investigated the contribution of the sympathetic nervous system (SNS) in maintaining the blood pressure and in regulating the cardiac function during and after carvedilol administration in rats with heart failure (group F). Left ventricular end-diastolic pressure, percent functional shortening, and rates of intraventricular pressure rise were significantly changed by carvedilol infusion as compared with the basal values in group N (normal rats), but not in group F. The left ventricular end-diastolic pressure was elevated, corresponding to the enhancement of the plasma norepinephrine (NE) concentration caused by carvedilol infusion, in group N. The enhancement of the plasma NE concentration induced by carvedilol administration in group F was higher than that in group N. The value for the maximal hypertensive effect of NE intravenous infusion (Emax) was decreased, and the plasma NE concentration at half-maximal effect (EC50) was increased in group F as compared with the values in group N. These results indicate that the SNS (presynaptic) activity is increased and that the SNS receptor sensitivity in the cardiovascular regulation system is decreased in heart failure. PMID- 15539881 TI - Influences of gender and region on responses to 5-HT in the rat small intestine. AB - The aim of this study was to obtain more information regarding the 'atypical' 5 HT7 receptor of the rat jejunum. 5-HT7-induced contractions of the jejunum were elicited by 5-HT in the presence of ondansetron. Maximal responses were slightly larger in tissues from male compared to female rats of comparable age, with Emax values of 97.2 +/- 3.3 and 84.25 +/- 4.3% respectively compared to acetylcholine as an internal standard. However, the pEC50 values for 5-HT were not significantly different. The mRNA expression levels of the 5-HT7 receptor were similar in whole jejunum and longitudinal muscle tissues taken from males and females. It was also shown that the maximal response of the jejunum from male rats was larger than the responses from mid intestine and ileum. However, in female tissues, the Emax of the mid intestine was significantly larger than the ileum, but not different from the jejunum. The results provide further insights into the 'atypical' 5-HT7 receptor of the rat jejunum and are also useful in optimising the preparation for further studies. PMID- 15539882 TI - Drug history overrides opioid reinforcement in a rat runway procedure. AB - To determine the effect of the drug history of rats on their subsequent operant behaviour in the rat runway procedure, we tested the mu opioid receptor agonists morphine and remifentanil and found a carryover of previous drug/saline experience that was not found for a food reinforcer. Previous exposure to saline significantly decreased the apparent reinforcing effect of subsequently offered morphine or remifentanil, while previous experience with morphine or remifentanil significantly increased responding for saline. This carryover of a previous learning experience on subsequent operant performance in the rat runway procedure cautions that the drug history should be considered and might even invalidate subsequent findings when testing drugs of abuse or other reinforcers. PMID- 15539883 TI - Relaxation induced by acetylcholine involves endothelium-derived hyperpolarizing factor in 2-kidney 1-clip hypertensive rat carotid arteries. AB - Acetylcholine induced relaxation in a concentration-dependent way in isolated phenylephrine-contracted carotid artery rings from normotensive two-kidney (2K) and hypertensive two-kidney one-clip (2K-1C) rats. In the presence of the nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine (L-NOARG, 100 micromol/l), the relaxation stimulated with acetylcholine was blocked in 2K arteries. However, in 2K-1C arteries, the relaxation was only partially inhibited. Indomethacin (3 micromol/l) had no effect in both groups. In 2K arteries, the combination of L NOARG and indomethacin had similar effects to L-NOARG alone. On the other hand, in 2K-1C arteries, indomethacin further inhibited the maximum effect induced by acetylcholine. Endothelium-dependent relaxation induced by acetylcholine was markedly reduced in 2K arteries contracted with 90 mmol/l KCl, and it was abolished in 2K-1C arteries. The remaining response to acetylcholine in 2K arteries was blocked by L-NOARG. Thus, in addition to NO, a relaxing factor sensitive to extracellular K+ changes in the membrane potential contributes to endothelium-dependent relaxation in 2K-1C rat carotid artery. On the other hand, in arteries from 2K rats, only NO is involved in the relaxation induced by acetylcholine. The combination of 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, 3 micromol/l), indomethacin (3 micromol/l) and L-NOARG (100 micromol/l) reduced the relaxation induced by acetylcholine in arteries from 2K-1C rats contracted with phenylephrine. On the other hand, in 2K arteries, the relaxation induced by acetylcholine was abolished. The combination of ODQ and K+ channel blockers charybdotoxin (100 nmol/l), apamin (500 nmol/l) and 4-aminopyridine (1 micromol/l) abolished the relaxation induced by acetylcholine in 2K and 2K-1C carotid arteries. These data indicate that the endothelium-derived relaxing factors that contribute to relaxation induced by acetylcholine are different in 2K and 2K-1C arteries. In 2K arteries, the only factor is NO, which involves the activation of K+ channels and the cGMP pathway. However, in 2K-1C arteries, the relaxation induced by acetylcholine is dependent on NO in addition to another factor, which is insensitive to indomethacin, but also activates the K+ channels and the cGMP pathway, presumably by membrane hyperpolarization through endothelium-derived hyperpolarizing factor. PMID- 15539884 TI - Effects of a novel anti-hyperlipidemic agent, S-2E, on blood lipid levels in rats with fructose-induced hypertriglyceridemia. AB - Using rats with fructose-induced hypertriglyceridemia, an animal model of human hypertriglyceridemia, we investigated whether (+)-(S)-p-[1-(p-tert-butylphenyl)-2 oxo-4-pyrrolidinyl]-methoxybenzoic acid (S-2E), a novel anti-hyperlipidemic agent, reduced the elevated levels of triglyceride (TG) and non-high-density lipoprotein cholesterol (non-HDL-C), and then whether it elevated HDL-C levels. At doses of 3-30 mg/kg, S-2E reduced elevated TG levels and non-HDL-C levels simultaneously in a dose-dependent manner after a week. Furthermore, S-2E treatment at 10 mg/kg for 4 weeks showed similar effects, while the elongation of intervals between feeding periods led to further increases in these levels. Interestingly, S-2E increased blood HDL-C levels after 4 weeks of treatment. It is therefore reasonable to assume that S-2E may be useful to improve dyslipidemia such as hypertriglyceridemia and low levels of HDL-C. PMID- 15539885 TI - Inhibition of rat intestinal Cl- secretion by 4,5-dimethylresorcinol. AB - Wood creosote, a mixture of phenolic compounds, inhibits enterotoxin-induced intestinal fluid secretion, suggesting that one of its constituents suppresses transepithelial Cl- secretion from the intestinal mucosa. To identify an active constituent in wood creosote that inhibits intestinal Cl- secretion through Cl- channels, we first examined its effect on Cl- secretion using a cultured cell line transfected with complementary DNA encoding a Cl- channel and a Cl- sensitive fluorescent dye. We next assayed chromatographic fractions of wood creosote for the inhibitory activity on Cl- secretion using a Ussing chamber. We found that 4,5-dimethylresorcinol, identified by gas chromatography-mass spectrometry, inhibited intestinal Cl- secretion dose-dependently when added to a serosal, but not mucosal, surface of rat jejunum, a half-inhibitory concentration being 3.8 microg/ml (28 micromol/l). It was strongly suggested that this effect was due to inhibition of Cl- channels. PMID- 15539886 TI - Clinical correlation between the consumption of nicotine and cotinine concentrations in urine and serum by competitive enzyme-linked immunosorbent assay. AB - Different negative effects of smoking are known (e.g. postoperative complications), which lead, especially in plastic surgery, to unsatisfactory results. The aim of this study was to examine the appropriateness of a cotinine enzyme-linked immunosorbent assay (ELISA) for routine usage in operative disciplines. By correlation of smoking habits and concentration of cotinine in serum and urine, we tried to ascertain reference values for smokers, passive smokers and non-smokers. The reliability and sensitivity of the cotinine ELISA concerning nicotine exposure need to be proven. 165 (108 men, 57 women) test persons were examined by detecting cotinine in serum and urine by ELISA. The study shows a very good sensitivity, precision and reproducibility of the cotinine ELISA according to the laboratory criteria. The test shows significance of a good differentiation between smokers, passive smokers and non-smokers in urine. In serum, cotinines are excellent to differentiate between smokers and non smokers/passive smokers. Urine and serum tests demonstrate valid and comparable results. PMID- 15539887 TI - Bilateral diagonal earlobe crease and coronary artery disease: a significant association. AB - BACKGROUND: The association between the presence of diagonal earlobe crease (ELC) and coronary artery disease (CAD) still remains controversial. OBJECTIVE: The aim of this study was to evaluate the association between bilateral ELC and CAD. METHODS: 415 patients were examined for the presence or absence of bilateral ELC, angiographic evidence of CAD and coronary risk factors. The patients were divided into 2 groups according to angiographic evidence of CAD. RESULTS: Bilateral ELC was significantly and positively correlated with CAD, hypertension, age, male gender, cigarette smoking and family history of CAD. The ELC was an independent variable for CAD. The observed sensitivity, specificity, positive predictive value and negative predictive value of the bilateral ELC for the diagnosis of CAD were in the following order: 51.3, 84.8, 89.4 and 41.2%. CONCLUSION: The presence of bilateral ELC was significantly associated with CAD and coronary risk factors. The bilateral ELC was an important dermatological indicator of CAD, and it might be a useful diagnostic tool in the clinical examination of patients. PMID- 15539888 TI - Is the atopy patch test with house dust mites specific for atopic dermatitis? AB - BACKGROUND: The atopy patch test (APT), namely the patch test with aeroallergens, is regarded as specific for patients with atopic dermatitis (AD), but small numbers of positive APT were reported in the past also in atopic subjects without dermatitis and in healthy persons. OBJECTIVE: The aim of this study was to evaluate the response to the APT with house dust mites (HDM) in subjects nonaffected by AD and to compare the outcomes observed in these cases with those pointed out in AD patients, evaluating also the differences between two allergen extracts manufactured at different purifications and concentrations. METHODS: Forty-seven atopic subjects without eczema (AWE), 33 nonatopic (NA) subjects and 77 adult AD patients were patch tested with an extract of purified bodies of HDM at 20% and with another extract of whole bodies of HDM at 30%, the latter corresponding to 300 microg/g of Der p 1. The reproducibility of APT was also tested in 8 AD patients, in 37 AWE subjects and in 19 NA subjects. RESULTS: Positive responses with extract at 20% were observed in 29 (37.7%) AD, in 5 (10.6%) AWE and in 4 (12.1%) NA subjects. The APT with HDM at 30% was positive in 32 (41.6%) AD, 9 (19.1%) AWE and 4 (12.1%) NA persons. The rates of positivity and the intensity scores of responses were significantly different between AD and non-AD subjects (p < 0.01). The reproducibility of the APT in the three groups was satisfactory. CONCLUSION: These observations lead to conclude that the APT with HDM is positive also in non-AD subjects but it is probably more specific for AD. PMID- 15539889 TI - Plexiform neurofibromas express the transcription factor Gli1. AB - BACKGROUND: Plexiform neurofibromas occur commonly in individuals with neurofibromatosis type 1 (NF1) and consist of neurofibromatous change in multiple nerve fascicles. Previously, we had observed that both plexiform neurofibromas and normal cutaneous nerves expressed Hedgehogs (Hhs), which are intercellular signaling molecules and regulate growth and patterning during embryonic development, and their receptors. In the present study, we examined the expression of Gli1, a transcription factor which mediates Hh signaling to investigate the activation of Hh signaling in plexiform neurofibromas and normal cutaneous nerves. METHODS: An antihuman Gli1 antibody was used with a standard immunoperoxidase technique to determine Gli1 expression in 5 specimens of plexiform neurofibromas and 5 specimens of normal cutaneous nerves. RESULTS: Our results showed Gli1 expression in S-100-positive tumor cells within the involved nerve fascicles in plexiform neurofibromas but not in control normal skins. CONCLUSIONS: Our findings indicate that the Hh signaling pathway is activated in plexiform neurofibromas. PMID- 15539890 TI - High nickel release from 1- and 2-euro coins: are there practical implications? AB - PURPOSE: To determine the release of nickel from 1- and 2-euro coins and the ability to produce allergic contact dermatitis from the application of coins to the palmar skin of nickel-sensitized individuals. METHODS: Three experiments were conducted. Experiments 1 and 2 checked the release of nickel from 1- and 2-euro coins by using the dimethylglyoxime test. In experiment 3, the elicitation of positive reactions was checked by applying coins to the palmar skin for 48 h under occlusion in nickel-sensitized and non-sensitized individuals. RESULTS: The dimethylglyoxime test for release of nickel was positive in all cases. Positive patch test reactions to euro coins applied to the palmar skin of nickel sensitized individuals were observed at 48 and 96 h. CONCLUSION: The results show that positive patch test reactions to euro coins can be obtained from nickel sensitized individuals after 48 h of application to the palmar skin under occlusion. These results do not contradict other experiments in which repeated handling of coins was unable to provoke fingertip allergic contact dermatitis. A dose-response relationship is a credible explanation to support such potential discrepancies. PMID- 15539891 TI - Idiopathic systemic capillary leak syndrome: cutaneous involvement can be misleading. AB - BACKGROUND: Systemic capillary leak syndrome (SCLS) is a severe disorder characterized by unexplained rapid transfer of considerable volumes of plasma from the intravascular to the extravascular compartment. For some cases of SCLS, no aetiology is evident and these cases are reported as idiopathic (ISCLS). OBJECTIVES: To describe the cutaneous findings in 3 patients with ISCLS. RESULTS: Cutaneous involvement consisted in sclerosis, livedo, purpura and photodistributed maculopapular erythematous rash. Dermal mucinosis was proven by biopsy in 1 patient. No underlying disease was diagnosed during follow-up. CONCLUSION: The above-mentioned cutaneous findings can be present during acute attacks of ISCLS. They seem specifically related to the ISCLS and not indicative of an underlying disease. PMID- 15539892 TI - Serological immunomarkers in cutaneous T cell lymphoma. AB - INTRODUCTION: As serological immunomarkers like neopterin, beta2-microglobulin, soluble IL-2 receptor (sIL-2R) and IL-6 have been described to be elevated in various malignancies, the aim of this study was to investigate whether they would be of diagnostic and prognostic value for leukemic and non-leukemic cutaneous T cell lymphoma (CTCL). PATIENTS AND METHODS: Forty-one CTCL patients from the lymphoma clinics of the Department of Dermatology, University of Zurich, were tested for the serum levels of the above-mentioned immunomarkers at several time points, and clinical status and clinical outcome were recorded. Thirty-nine patients with CBCL and T cell inflammatory diseases served as controls. RESULTS: The study revealed that neopterin, beta2-MG and sIL-2R are significantly elevated in Sezary syndrome, whereby sIL-2R seemed to be the most sensitive marker and is typically increased in Sezary syndrome. Moreover, there is a correlation between tumor burden index values and serum parameters. Concerning the outcome of the disease (progression versus non-progression), only neopterin showed a significant prognostic value in non-leukemic CTCL patients. CONCLUSION: Serological immunomarkers are helpful tools in determining the tumor burden in CTCL and thus might be useful for disease monitoring during treatment. They may have prognostic value for predicting the clinical course. PMID- 15539893 TI - Chronic dermatomycoses of the foot as risk factors for acute bacterial cellulitis of the leg: a case-control study. AB - OBJECTIVE: To assess the role of foot dermatomycosis (tinea pedis and onychomycosis) and other candidate risk factors in the development of acute bacterial cellulitis of the leg. METHODS: A case-control study, including 243 patients (cases) with acute bacterial cellulitis of the leg and 467 controls, 2 per case, individually matched for gender, age (+/-5 years), hospital and admission date (+/-2 months). RESULTS: Overall, mycology-proven foot dermatomycosis was a significant risk factor for acute bacterial cellulitis (odds ratio, OR: 2.4; p < 0.001), as were tinea pedis interdigitalis (OR: 3.2; p < 0.001), tinea pedis plantaris (OR: 1.7; p = 0.005) and onychomycosis (OR: 2.2; p < 0.001) individually. Other risk factors included: disruption of the cutaneous barrier, history of bacterial cellulitis, chronic venous insufficiency and leg oedema. CONCLUSIONS: Tinea pedis and onychomycosis were found to be significant risk factors for acute bacterial cellulitis of the leg that are readily amenable to treatment with effective pharmacological therapy. PMID- 15539894 TI - Efficacy of treatment with calcipotriol/betamethasone dipropionate followed by calcipotriol alone compared with tacalcitol for the treatment of psoriasis vulgaris: a randomised, double-blind trial. AB - BACKGROUND: A two-compound product containing calcipotriol 50 microg/g and betamethasone dipropionate 0.5 mg/g (Daivobet, Dovobet) has been demonstrated to be an effective, once daily, treatment for psoriasis vulgaris. OBJECTIVE: To compare the efficacy and safety of treatment with the two-compound product for 4 weeks followed by calcipotriol for 4 weeks, with that of tacalcitol for 8 weeks in patients with stable psoriasis vulgaris. METHODS: 501 patients were randomised to double-blind treatment with the two-compound product followed by calcipotriol 50 microg/g once daily, or to tacalcitol 4 microg/g once daily. RESULTS: Treatment with the two-compound product/calcipotriol was significantly more effective than tacalcitol in terms of mean percentage PASI reduction (65.0 vs. 33.3% at week 4 and 59.0 vs. 38.4% at week 8; p < 0.001 for both). CONCLUSION: A treatment regimen comprising calcipotriol/betamethasone ointment (Daivobet) for 4 weeks followed by calcipotriol for 4 weeks is superior to tacalcitol ointment for 8 weeks in patients with psoriasis vulgaris. PMID- 15539895 TI - Influence of pimecrolimus cream 1% on different morphological signs of eczema in infants with atopic dermatitis. AB - BACKGROUND: In the published studies on the efficacy of the topical immunomodulator pimecrolimus, different eczema scores were used, and the impact on morphological key signs of eczema was not analysed. OBJECTIVE: To compare the influence of pimecrolimus cream 1% on different standard eczema scores in infants with atopic dermatitis and to analyse the impact of treatment on the individual morphological key signs of eczema. METHODS: Pimecrolimus cream 1% (n = 129) or double-blind vehicle control (n = 66) was administered for 4 weeks. The Eczema Area and Severity Index (EASI), Investigators' Global Assessment (IGA) and Scoring Atopic Dermatitis Index (SCORAD) were determined and were correlated with each other. RESULTS: Following treatment with pimecrolimus, the EASI, IGA and SCORAD were significantly reduced on day 29 as compared with the vehicle group (p < 0.001, p < 0.001, p = 0.002, respectively). There was a close correlation between EASI, IGA and SCORAD. The single parameters of the EASI were already significantly decreased by day 4 in the pimecrolimus group as compared to vehicle (each p < 0.001). CONCLUSION: Treatment with pimecrolimus 1% cream leads to a rapid improvement of all morphological signs of eczema. The close correlation of different scores was shown for the first time. PMID- 15539896 TI - Partial unilateral lentiginosis with ipsilateral Lisch nodules and axillary freckling. AB - A 9-year-old Taiwanese girl had an unusual combination of multiple lentigines clustered on her left face, 3 cafe-au-lait spots on her left arm and ipsilateral axillary freckling. Two Lisch nodules of the iris were recognized in the left eye but none in the right eye, which is why we established a diagnosis of segmental neurofibromatosis 1. No family member had any sign of neurofibromatosis 1. Moreover, the patient had a large speckled lentiginous nevus involving the left side of her trunk, and her father showed a similar skin disorder on his left buttock. These lesions were taken to be unrelated to the patient's segmental neurofibromatosis 1. This case provides further evidence in favor of the concept that partial unilateral lentiginosis represents a mosaic manifestation of type 1 neurofibromatosis. PMID- 15539897 TI - Successful treatment of recalcitrant chronic actinic dermatitis with tacrolimus. AB - Therapeutically, chronic actinic dermatitis is a problematic condition. Frequently applied sunscreen usually fails to mitigate the clinical symptoms, and steroids--while efficient--exert many undesired side-effects with prolonged usage. We present a case of chronic actinic dermatitis responding well to topically applied tacrolimus (Protopic) in combination with chemical and physical UV protection. PMID- 15539898 TI - Elevated amount of human papillomavirus 31 DNA in a squamous cell carcinoma developed from bowenoid papulosis. AB - A 50-year-old woman presented with asymptomatic, multiple black macules on the genitalia for 6 months. She presented multiple, round to irregularly shaped, black macules on her labia major and perineum. A keratotic reddish nodule was found on one of the macules. The results of laboratory investigations showed pancytopenia and a decreased CD4/CD8 ratio. A lymphocyte stimulation test showed a decreased response. The histological examination of a biopsy specimen led us to the diagnosis of bowenoid papulosis (BP) for the macules and squamous cell carcinoma (SCC) for the nodule. The macules and nodule were removed successfully. An elevated amount of HPV DNA was demonstrated in SCC compared with that of BP. The HPV DNA in SCC and BP was shown to be HPV-31. These observations indicated that the elevated amount of HPV-31 DNA could have led to the emergence of SCC from BP under the condition of decreased cellular immunity. PMID- 15539899 TI - Faun tail with aplasia cutis congenita and diastematomyelia. PMID- 15539900 TI - Sclerosing cholangitis and liver transplantation in Langerhans cell histiocytosis: a 14-year follow-up. PMID- 15539901 TI - Vellous hair in trichoepithelioma papulosum multiplex. PMID- 15539902 TI - Magnetic resonance imaging of semicircular lipoatrophy. PMID- 15539903 TI - Increased expression of segmental neurofibromatosis with bronchoalveolar lung carcinoma. PMID- 15539904 TI - Localized urticaria pigmentosa triggered by mesotherapy. PMID- 15539905 TI - Psychodermatoses in haemodialysis patients. PMID- 15539906 TI - Fulminant streptococcal necrotizing fasciitis. PMID- 15539909 TI - Nuclear receptors and unsaturated fatty acids: implications for pharmacologic activity of the galenic forms of topical preparations. PMID- 15539910 TI - Diffuse melanosis cutis in disseminated malignant melanoma. PMID- 15539911 TI - Combined irinotecan and oxaliplatin in patients with advanced pre-treated pancreatic cancer. AB - OBJECTIVES: This study evaluated the clinical activity and toxicity of combination chemotherapy with irinotecan and oxaliplatin in patients with advanced pancreatic cancer that had progressed despite > or =1 course of a gemcitabine-containing regimen. METHODS: Thirty patients with metastatic pancreatic cancer and Karnofsky performance status > or =70 received oxaliplatin 60 mg/m2 on days 1 + 15 and irinotecan 60 mg/m2 on days 1 + 8 + 15 every 4 weeks. Patients were assessed on the basis of clinical benefit response, changes in serum tumour marker CA 19-9, objective tumour response, time to progressive disease (TTP), and survival. RESULTS: Six patients (20%) had clinical benefit response (median duration of 7.2 months). CA 19-9 levels were reduced > or =50% from baseline in 8 patients (26%) and remained stable in 8 patients. CT scans revealed that 3 patients (10%) had a partial response and 7 (23%) had stable disease. Two patients (7%) were down-staged and underwent surgery. Median TTP was 4.1 months, median survival was 5.9 months and the 1-year survival rate was 23.3%. The most serious adverse events were grade 3-4 leukopenia in 2 patients (6%), grade 3 neuropathy in 2 (6%) and grade 3 diarrhoea in 1 (3%). CONCLUSION: Chemotherapy with irinotecan and oxaliplatin is an active and well-tolerated combination in patients with advanced pre-treated pancreatic cancer. PMID- 15539912 TI - Clinicopathological and biological characteristics of esophageal squamous cell carcinoma associated with head and neck cancer. AB - OBJECTIVE: Esophageal cancer has been reported to be frequently associated with cancer of the head and neck. The iodine dye method is reportedly useful to detect early esophageal cancer. The aim of this study was to clarify clinicopathological and biological characteristics of esophageal squamous cell carcinoma associated with head and neck cancer (HN group). METHODS: Thirty-seven patients of the HN group who underwent esophagectomy were examined clinicopathologically compared to 42 patients with esophageal cancer alone (SE group). All resected specimens were histologically studied after iodine dye staining, and p53 and cyclin D1 (CD1) expression were immunohistochemically examined in esophageal cancer. RESULTS: The HN group had more multiple iodine-unstained lesions and multiple primary cancers within the esophagus compared with the SE group (p = 0.0027, p = 0.067, respectively). There was no significant difference in smoking, drinking, family history and the other clinicopathological factors between the HN and SE groups. Coexpression of p53 and CD1 was found to be significant in the HN group (p = 0.03). CONCLUSIONS: The coexistence of multiple iodine-unstained lesions, multiple cancers in the esophagus and overexpression of p53 and CD1 is suggested as a risk factor for the HN group. PMID- 15539914 TI - Safety of long-term administration of bisphosphonates in elderly cancer patients. AB - BACKGROUND: To evaluate the effectiveness and tolerability of the long-term treatment bone metastases with pamidronate in older patients. MATERIALS AND METHODS: Twenty-two ambulatory patients aged 70 or older were included in the study. The median age was 73 (range 70-77). Ten patients (46%) were affected by breast carcinoma, 7 (32%) by prostate carcinoma and 5 (22%) by multiple myeloma. Nine (40%) patients presented co-morbidity. All of the patients presented at least one metastatic lytic bone lesion measuring 1 cm or more in diameter; the median lesion number was 2 (range 1-4). Hormonal therapy or chemotherapy regimen, were allowed as clinically required. Patients were treated with a fixed dose of sodium pamidronate, 90 mg in 3 h infusion every 4 weeks. RESULTS: Partial response was shown in 6 (28%) patients, stable disease in 11 (50%), and progression (PD) in 5 (22%). 2 out of 5 patients with PD presented skeletal related events (SREs) such as bone fracture. The median treatment duration was 19 months. The treatment was well tolerated; in 5 patients (23%) a GI fever was observed, in 3 patients (18%) G1 nausea, and in 3 patients (14) G1 diarrhea. Two cases (9%) of acute renal insufficiency (creatinine 1.7 and 1.6 mg/dl), and 3 cases (14%) of hypocalcemia (7.6, 7.5 and 7.8 mg/dl) were also registered. The renal dysfunction was reversible and without consequence. CONCLUSION: Our experience suggests that the bisphosphonates long-term administration is useful and did not cause significant side effects in elderly subjects. Low-grade pyrexia, nausea/vomiting, acute/reversible renal dysfunction and hypo-calcemia were the most frequent side effects reported. However, they were of low grade and in most cases, did not require dose modifications and/or hospitalization. PMID- 15539913 TI - Concomitant radiochemotherapy plus surgery in locally advanced cervical cancer: update of clinical outcome and cyclooxygenase-2 as predictor of treatment susceptibility. AB - OBJECTIVE: We have updated our findings on the efficacy of concomitant radiochemotherapy plus radical surgery in a larger series of patients (n = 54) with locally advanced cervical cancer (LACC). We also investigated the role of cyclooxygenase-2 (COX-2) in this clinical setting. METHODS: Radiotherapy was administered to the whole pelvic region (1.8 Gy/day, totaling 39.6 Gy) in combination with cisplatin (20 mg/m2) and 5-fluorouracil (1,000 mg/m2) (both on days 1-4 and 27-30). Radical surgery was performed 5-6 weeks after the end of treatment. RESULTS: A clinical complete or partial response was observed in all 53 evaluable patients (75.5 and 24.5%, respectively). At pathological examination, 23 of 51 patients (45.1%) undergoing radical surgery showed complete response to treatment, 18 patients (35.3%) only had microscopic residual disease, 6 patients (11.7%) had a partial response and 4 (7.8%) had no change in their disease. When logistic regression was applied, the FIGO stage (chi2 = 5.28, p = 0.021) and tumor to stroma COX-2 ratio (chi2 = 4.72, p = 0.029) retained an independent role in the prediction of the pathologic response to treatment. The 3 year disease-free survival (DFS) was 75.2%, with local relapse-free survival of 86.2% and metastasis-free interval of 89.9% at 3 years. Cases with a high COX-2 ratio showed a shorter DFS than cases with a low COX-2 ratio (p = 0.016). A direct association was shown between COX-2 ratio values and risk of recurrence, as assessed by Cox analysis using COX-2 ratio values as a continuous covariate (chi2 = 3.94, p = 0.047). CONCLUSION: This study confirms the possibility of achieving a very high rate of pathological responses in LACC patients administered chemoradiation plus surgery (3-year DFS 75.2%). Moreover, COX-2 status may play a role in the prognostic characterization and prediction of tumor response. PMID- 15539915 TI - Capecitabine with weekly paclitaxel for advanced breast cancer: a phase I dose finding trial. AB - OBJECTIVE: To determine the maximum tolerated dose (MTD), toxicity and activity of combined weekly paclitaxel and capecitabine in patients with metastatic breast cancer. METHODS: Sixteen patients with metastatic breast cancer, of whom 15 were evaluable for toxicity and response, were enrolled in 7 Swiss centers. Paclitaxel 80 mg/m2 was given intravenously on days 1, 8 and 15. Capecitabine was administered orally on days 1 through 14 using five different dose levels. Both drugs were given in a 21-day schedule. RESULTS: Capecitabine could be administered at doses commonly used for the drug as a single agent, i.e. 1,250 mg/m2 twice daily in combination with weekly paclitaxel. Hematological and other toxicities did not appear to be dose-limiting; however, significant skin and nail toxicities were observed. A response or stable disease was observed in 87% of patients [13/15; exact 95% confidence interval (CI) 60-98%], with 2 complete responses, 4 partial responses (overall response rate 40%, exact 95% CI 16-68%) and 7 patients with stable disease for at least 9 weeks. CONCLUSION: The phase I evaluation of capecitabine in combination with fixed-dose weekly paclitaxel did not allow the definition of an MTD of capecitabine based on the predefined criteria. Instead, the dose for the phase II evaluation was determined based on the occurrence of toxicity in later courses and on experience with other regimens containing capecitabine. Capecitabine (1,000 mg/m2 twice daily, days 1-14, every 3 weeks) with paclitaxel (80 mg/m2 weekly) is a promising combination for advanced breast cancer now being investigated in a phase II trial. PMID- 15539916 TI - A phase I and pharmacokinetic study of BAY59: a novel taxane. AB - PURPOSE: To determine the maximum tolerated dose (MTD), the dose limiting toxicities (DLT) and the pharmacokinetics of BAY59, a novel taxane given as a 1 hour intravenous infusion every 3 weeks in patients with advanced refractory solid tumors. EXPERIMENTAL DESIGN: Initially, 15 patients with previously treated (median of 4 prior chemotherapy regimens) refractory cancers, but with normal marrow, hepatic and renal function were treated with BAY59 at doses of 15, 30, 50, 75 and 100 mg/m2 using a standard dose escalation design. Subsequently, 11 patients were treated, 5 at 90 mg/m2 and 6 who had had prior oxaliplatin at 75 mg/m2. RESULTS: At 75 mg/m2, grade 4 neutropenia was noted in 2/6 patients, of whom 1 had grade 4 neutropenia lasting more than 5 days (DLT). At 100 mg/m2, 2/2 patients had febrile neutropenia, with 1 fatality. At 90 mg/m2, 2/5 patients had DLTs, including grade 3 neuropathy, severe lower extremity pain, dehydration and grade 4 neutropenia. The MTD was determined to be 75 mg/m2. A cohort of 6 patients, previously exposed to oxaliplatin, were enrolled at the MTD to evaluate the incidence of neurotoxicity. While DLTs (grade 3 arthralgia, grade 4 neutropenia) were noted in 3/6 patients, there was no increase in the incidence of neurotoxicity. There were no responses. Pharmacokinetics of BAY59 was linear over the doses studied, with a median terminal half-life of 21 h. CONCLUSIONS: The recommended phase II dose for BAY59 is 75 mg/m2. PMID- 15539917 TI - Fas gene mutations in mycosis fungoides: analysis of laser capture-microdissected specimens from cutaneous lesions. AB - Fas (APO-1/CD95) is a transmembrane protein which mediates programmed cell death (apoptosis). Cells with a mutated Fas gene are resistant to apoptosis and thus accumulate in lesional tissues. This might provide a basis for the development of neoplasias. Genomic DNA selectively obtained from Pautrier's microabscesses in 16 cases of mycosis fungoides (MF) using a laser capture microdissection method was analyzed. Fas gene mutations were detected in 3 of 16 cases of MF (18.8%); 1 was silent and 2 were missense mutations located in exon 9. One of the 2 missense mutations involved the death domain of the Fas gene, which is essential for apoptotic signal transduction. The missense mutations resulted in the substitution of Ala with Asp at codon 220 and Ile with Thr at codon 314. Mouse T cell lymphoma cells transfected with mutant genes were resistant to apoptosis induced by the anti-Fas antibody, indicating that the missense mutations found in MF were loss-of-function mutations, thus causing the accumulation of cells in the cutaneous lesions. These findings suggest that the accumulation of lymphoid cells with Fas mutations provides, in part, a basis for the development or maintenance of MF. PMID- 15539919 TI - Prognostic significance of serine 392 phosphorylation in overexpressed p53 protein in human esophageal squamous cell carcinoma. AB - OBJECTIVE: Posttranscriptional modification involving phosphorylation of wild type p53 has been considered to play a role in the stabilization of p53. Little is known, however, about the role of phosphorylation of mutant p53 overexpressed in tumors. The aim of this study is to determine the phosphorylation state of p53 in tumors and its contribution to tumor development. METHODS: Using immunohistochemical techniques, we examined the phosphorylation of Ser392 and 15 sites in p53 overexpressed in 137 esophageal squamous cell carcinomas (ESCCs). The relationships between the phosphorylation and the expression of cell cycle regulators, Ki-67 labeling index (LI), apoptotic index, clinicopathological factors, and patient prognosis were tested statistically. RESULTS: Of the 137 samples examined, staining for Ser392 phosphospecific p53 antibody was detected in 53 (38.7%) cases, corresponding to 58.9% of 90 p53-positive cases, whereas only 3 (2.2%) were positive for Ser15 phosphospecific p53 antibody. A significant correlation was identified between Ser392 phosphorylation and high levels of Ki 67 LI (p = 0.0271), lymphatic invasion (p = 0.0246), and poorer prognosis for patients with stage II and III advanced tumors (p = 0.0136). Using multivariate analysis, Ser392 phosphorylation was recognized as an independent prognostic factor (p = 0.0136). CONCLUSIONS: These findings suggest that p53 protein overexpressed in ESCCs is frequently phosphorylated at Ser392, and that the Ser392 phosphorylation might contribute to tumor progression of ESCCs. PMID- 15539920 TI - Frequent loss of heterozygosity but rare microsatellite instability in oesophageal cancer in Japanese and Chinese patients. AB - Reported frequencies for microsatellite instability (MSI) in oesophageal cancer differ widely in the literature, perhaps due to the high incidence of loss of heterozygosity (LOH) in this cancer. Using high-resolution fluorescent microsatellite analysis (HRFMA), we analysed microsatellite alterations in detail in 50 Japanese and 50 Chinese patients with squamous cell carcinoma in the oesophagus. In HRFMA, several devices have been developed to improve the detection characteristics, reproducibility of polymerase chain reaction and the migration accuracy of electrophoresis. All the alterations observed were separable into MSI, LOH and alterations ambiguous for both. MSI was rare in these panels of oesophageal carcinomas. The frequencies of MSI in the Japanese and Chinese subjects were 8 and 4%, respectively. All the alterations were mild (within 2 base pairs) and were observed in a limited number of markers. More drastic types of MSI, such as those typical in colorectal cancer, were not observed. On the other hand, the incidence of LOH was high, reaching 50% for the Japanese and 70% for the Chinese subjects. In many of these cases, LOH was observed in multiple microsatellite markers. The frequency of LOH in each marker was not apparently biased. Although in many cases MSI and LOH were clearly distinguished with use of the sensitive and quantitative fluorescent assay, theoretically indistinguishable patterns were noted in some cases. In conclusion, MSI is rare and LOH predominates in squamous cell carcinoma in the oesophagus. PMID- 15539918 TI - Thymidylate synthase expression in normal colonic mucosa: a predictive marker of toxicity in colorectal cancer patients receiving 5-fluorouracil-based adjuvant chemotherapy. AB - OBJECTIVES: We retrospectively evaluated the relevance of thymidylate synthase (TS) expression in normal colonic mucosa as a predictive factor of toxicity in colorectal cancer patients receiving adjuvant fluorouracil (5-FU)-based chemotherapy. METHODS: TS expression was immunohistochemically assessed on normal colonic mucosa from 50 patients with colorectal cancer Dukes' stages B (15 patients) and C (35 patients) treated with 5-FU-based adjuvant chemotherapy. RESULTS: The incidence of grade 2-3 diarrhea and stomatitis (according to WHO) was demonstrated to be significantly higher in patients with low nuclear TS expression in normal mucosa than in those with high TS expression (p < 0.0001). Moreover, patients with low TS expression in normal colonic mucosa developed weight loss and worsening of the performance status (according to ECOG score) more commonly than patients with high TS expression (p = 0.005 and p = 0.02, respectively). Furthermore, low TS expression in normal colonic mucosa significantly correlated with a higher rate of a delay of chemotherapy courses, dose reduction and treatment discontinuation. CONCLUSIONS: Immunohistochemical TS expression in normal colonic mucosa may represent an important predictive parameter for identifying a subset of patients with a high risk of developing severe 5-FU-related toxicities. PMID- 15539921 TI - Correlation analysis between tumor-associated antigen sialyl-Tn expression and ST6GalNAc I activity in human colon adenocarcinoma. AB - OBJECTIVES: Sialyl-Tn (sTn) is a mucin carbohydrate-associated antigen that is strongly expressed in a large number of colorectal carcinomas. In this study, we combined immunohistochemical and enzymatic techniques in order to find the correlation between sTn tissue expression and the sialyltransferase activity (ST6GalNAc I) responsible for its synthesis in colorectal cancer (CRC) patients. METHODS: We compared sTn expression in healthy (n = 46), tumorous (n = 60) and transitional tissue (n = 46) from CRC patients, and correlated sTn altered expression with clinicopathologic variables of the patient. Furthermore, we determined ST6GalNAc I tissue activity employing asialo-ovine submaxillary mucin (asialo-OSM) as glycoprotein acceptor (n = 27). RESULTS: The rates of sTn positive expression obtained for healthy, tumorous and transitional tissues were 15, 67 and 63%, respectively. These rates led to statistically significant differences between healthy and tumorous or transitional tissue (p = 0.001); sTn expression was related to the first stages of the tumor invasion in transitional tissue. As regards ST6GalNAc I activity, we found an enhancement in transitional tissue. Statistical correlation analysis did not reveal association between sTn expression and ST6GalNAc I activity. CONCLUSIONS: Our findings indicated that sTn antigen tissue expression and ST6GalNAc I activity levels were not correlated in CRC, in spite of the overexpression of the antigen in tumorous and transitional tissue. PMID- 15539922 TI - Thrombin inhibitor, argatroban, prevents tumor cell migration and bone metastasis. AB - It is well known that malignant cells show procoagulant activity, which is associated with their metastatic potential. Thrombin, the key enzyme of the blood coagulation system, is generated around tumor cells, promoting the migration and metastasis of tumor cells. In this study, we evaluated the effect of argatroban, a specific thrombin inhibitor, on the migration and metastasis of B16BL6 melanoma cells. In vitro argatroban dose-dependently inhibited cell migration, the maximum inhibition being observed in the presence of 10 microM argatroban (p < 0.0001). In order to investigate the antimetastatic effect of the thrombin inhibitor, we used an animal model that we have reported previously. C57BL6 mice which had received a bone (femur or tibia) transplanted into the dorsal subcutis were injected with B16 melanoma cells into the left heart ventricle. Intraperitoneal injection of argatroban (9 mg/kg/day for 4 weeks) significantly reduced the number of limbs with metastatic lesions as compared to a placebo (p < 0.05). These results suggest that argatroban was associated with reduced melanoma metastases by inhibiting cell migration. Our results showed that argatroban is effective for treatment of bone metastasis. PMID- 15539923 TI - Brain metastasis responding to gefitinib alone. AB - A woman with stage IIIb non-small cell lung cancer (NSCLC) developed disease progression with brain metastases during chemotherapy. Due to unusual circumstances, the patient received gefitinib alone, without the use of corticosteroid treatment or radiotherapy. There was a dramatic clinical improvement within 1 week. Follow-up magnetic resonance imaging of the brain 1 month later showed decreases in both the size and number of brain metastases. The patient remains well 9 months after initiation of gefitinib. It is proposed that gefitinib may have a role in treatment of brain metastases from NSCLC. PMID- 15539924 TI - Oxaliplatin-induced acute-onset thrombocytopenia, hemorrhage and hemolysis. AB - BACKGROUND: Oxaliplatin is a novel platinum derivative with established anti tumor activity in colorectal cancer. Acute-onset hemolytic anemia and thrombocytopenia associated with this drug have rarely been reported and some of these cases have been severe or even fatal. CASE REPORT: This case report describes a patient who developed fever, chills, abdominal and back pain as well as sudden-onset severe thrombocytopenia, upper gastrointestinal bleeding and hemolysis immediately after treatment with oxaliplatin for metastatic colorectal cancer. The reaction appeared during the 14th cycle of chemotherapy. Corticosteroids and antihistamines were administered together with platelet transfusions. Over the next 2 days platelet count improved and the syndrome abated. The patient was discharged 4 days later. Furthermore, the reaction was accompanied by a strongly positive Coombs test and increased TNF-alpha and IL-10 serum levels which returned to normal following anti-inflammatory drug administration. CONCLUSION: Physicians should be aware of the possibility of acute hematological emergencies following oxaliplatin administration. PMID- 15539925 TI - Insights into the taxonomy, genetics and physiology of bifidobacteria. AB - Despite the generally accepted importance of bifidobacteria as probiotic components of the human intestinal microflora and their use in health promoting foods, there is only limited information about their phylogenetic position, physiology and underlying genetics. In the last few years numerous molecular approaches have emerged for the identification and characterization of bifidobacterial strains. Their use, in conjunction with traditional culturing methods, has led to a polyphasic taxonomy which has significantly enhanced our knowledge of the role played by these bacteria in the human intestinal ecosystem. The recent adaptation of culture-independent molecular tools to the fingerprinting of intestinal and food communities offers an exciting opportunity for revealing a more detailed picture of the true complexity of these environments. Furthermore, the availability of bifidobacterial genome sequences has advanced knowledge on the genetics of bifidobacteria and the effects of their metabolic activities on the intestinal ecosystem. The release of a complete Bifidobacterium longum genome sequence and the recent initiative to sequence additional strains are expected to open up a new era of comparative genomics in bifidobacterial biology. Moreover, the use of genomotyping allows a global comparative analysis of gene content between different bifidobacterial isolates of a given species without the necessity of sequencing many strains. Genomotyping provides useful information about the degree of relatedness among various strains of Bifidobacterium species and consequently can be used in a polyphasic identification approach. This review will deal mainly with the molecular tools described for bifidobacterial identification and the first insights into the underlying genetics involved in bifidobacterial physiology as well as genome variability. PMID- 15539926 TI - Modulation of glycogen and trehalose levels in Micromonospora echinospora (ATCC 15837). AB - The growth of Micromonospora echinospora was studied in high and low C/N ratio medium using both batch and continuous culture. Asparagine was consumed rapidly in batch cultures where it served as both a nitrogen and carbon source. Glucose consumption was low suggesting that asparagine functions as the major carbon source under these conditions. The effect of nutrient limitation on the accumulation of storage carbohydrate in batch culture revealed an intimate association between nitrogen limitation and the accumulation of carbonaceous reserves. This study revealed that glycogen constituted the major carbohydrate reserve associated with the onset of sporulation. Intracellular trehalose levels were found to be relatively low and may have been affected by the availability of carbon. Continuous culture studies revealed a correlation between glycogen accumulation and increasing growth rate. It was also found that elevated cellular ATP levels correlated with the increase in glycogen, and reduced glycolytic activity. At the higher growth rates cellular ATP levels were elevated and coincided with reduced activity of the key glycolytic enzyme, phosphofructokinase, suggesting that glycogen can act as a convenient energy reservoir when excess carbon flux dictates. PMID- 15539927 TI - Nutrient utilization profile of Saccharomyces Cerevisiae from palm wine in tropical fruit fermentation. AB - The nutrient utilization pattern of Saccharomyces cerevisiae from palm wine was studied using tropical fruits as substrate. Starter cultures were prepared by growing 15-18 h old stock cultures of the yeast in successively larger bottles containing pasteurized fruit must. Microvinification, substrate utilization and assay of yeast activity were performed. Soluble solute (SS) content of the juices ranged from 10-18 Brix. Pinapple must had the highest SS content (18 Brix) while pawpaw had a low SS value of 10 Brix. These SS values were low compared to that of grape juice. The wines produced from the fruit must had percentage alcohol levels ranging from 10.6 to 12.6. Volatile activity ranged from 0.25 to 0.32 while crude protein values ranged from 0.58 to 0.68%. Palm wine yeast and all the other yeast strains fermented and utilized the fruit must for growth with specific growth rates ranging from 0.18 to 0.22. Sugar loss in Brix was gradual for all the fruit musts from 20.0-24.0 Brix to a range of 4.8 to 6.0 Brix. Pineapple was highly preferred for tropical wine making. Mango, cashew and pawpaw had equal ranking for commercial scale fermentation though more sugar will be needed to ameliorate cashew and pawpaw than mango juice. Palm wine yeast (OW-11) compared favourably with the other wine yeasts (CBS 8066 and ATCC 4126) both in nutrient utilization pattern and growth performance. A high degree of adaptability was observed in palm wine yeast recommands it for industrial wine production. PMID- 15539929 TI - Influence of host plant-derived and abiotic environmental parameters on the composition of the diazotroph assemblage associated with roots of Juncus roemerianus. AB - Environmental factors governing the distributions of plant root-associated bacteria are poorly understood. Most plant species occurring in salt marsh estuaries are restricted to very specific habitats within the marsh and plant derived and abiotic environmental features covary. We examined diazotrophic bacteria inhabiting the rhizoplanes of different populations of the black needlerush, Juncus roemerianus , growing in two different habitats, in order to examine the relative influence of plant-derived and abiotic environmental parameters on diazotroph assemblage composition. Juncus roots were collected from a monotypic Juncus patch in the low intertidal marsh, and from the main monotypic Juncus stand in the high marsh. A total of 235 bacterial pure cultures were isolated from the roots using combined nitrogen-free media. Physiologically similar strains were grouped, producing 58 different groups. Strains representing 49 of these groups tested positive for nifH , and substrate utilization profiles of these strains were compared quantitatively. Three major substrate utilization clusters were identified and all contained both Juncus patch and main stand isolates. Denaturing gradient gel electrophoresis analysis of nifH amplicons recovered from roots and from vegetated sediments taken from the main stand and from two patches was also performed. Juncus root nifH amplicon profiles from all three sampling sites were very similar. Profiles of amplicons from vegetated sediments were also similar across sites, but less similar than the root profiles. Results from two independent methodological approaches indicated a strong impact of the plant host relative to that of the abiotic environment on the composition of the root-associated diazotroph assemblage. PMID- 15539928 TI - Pectinatus portalensis nov. sp., a relatively fast-growing, coccoidal, novel Pectinatus species isolated from a wastewater treatment plant. AB - The genus Pectinatus is currently composed by two species, Pectinatus cerevisiiphilus and Pectinatus frisingensis , both asociated with beer spoilage. This study describes a novel isolate (strain B6) retrieved from a wastewater treatment plant collecting residues from a large number of wineries. Based on similarity analysis of 16S rRNA gene sequences, strain B6 belongs to the genus Pectinatus . Strain B6 is a strict anaerobe like other Pectinatus species and it presents non-motile, coccoid cells showing a slight oval shape. Strain B6 shows marked physiological differences with other Pectinatus species both in fatty acid composition and carbon source utilization. The most abundant fatty acids found in strain B6 were 18:1 (42.8%) and 16:0 (18.3%) representing a total of over 61% of fatty acids in this microorganism while these fatty acids represented 41.3% in P. cerevisiiphilusT and 2.4% in P. frisingensisT of their total. Fatty acid 15:0 was not significant in strain B6 and represented 28.6% and 13.3% for P. cerevisiiphilusT and P. frisingensisT, respectively. Strain B6 showed a faster growth rate and higher optimum temperature than its relatives P. cerevisiiphilus and P. frisingensis . Strain B6, P. cerevisiiphilus and P. frisingensis could be clearly differentiated by acid production tests from substrates such as esculine and gluconate, and the lack of acid production from rhamnose and fucose among others. G+C mol% content in strain B6 is 36.5%. Based on genotypic and phenotypic differences, strain B6 is proposed as a novel Pectinatus species, P. portalensis nov. sp. Both strain B6 and the two described species of Pectinatus grow on beers and wines. These results provide insights about the origin and reservoirs of Pectinatus species and spoiling alcoholic beverages. PMID- 15539930 TI - 16S rDNA library-based analysis of ruminal bacterial diversity. AB - Bacterial 16S rDNA sequence data, incorporating sequences > 1 kb, were retrieved from published rumen library studies and public databases, then were combined and analysed to assess the diversity of the rumen microbial ecosystem as indicated by the pooled data. Low G+C Gram positive bacteria (54%) and the Cytophaga Flexibacter-Bacteroides (40%) phyla were most abundantly represented. The diversity inferred by combining the datasets was much wider than inferred by individual studies, most likely due to different diets enriching for bacteria with different fermentative activities. A total of 341 operational taxonomic units (OTU) was predicted by the Chao1 non-parametric estimator approach. Phylogenetic and database analysis demonstrated that 89% of the diversity had greatest similarity to organisms which had not been cultivated, and that several sequences are likely to represent novel taxonomic groupings. Furthermore, of the 11% of the diversity represented by cultured isolates (> 95% 16S rDNA identity), not all of the bacteria were of ruminal origin. This study therefore reinforces the need to reconcile classical culture-based rumen microbiology with molecular ecological studies to determine the metabolic role of uncultivated species. PMID- 15539931 TI - Shewanella putrefaciens in a fuel-in-water emulsion from the Prestige oil spill. AB - Microorganisms that colonize the fuel-in-water emulsion from the Prestige spill have been compared with those from Exxon-Valdez. Both emulsions contained non fermentative gram-negative rods but unlike Exxon-Valdez's, the Prestige's spill contained anaerobic bacteria and no fungi. Our main finding has been the identification of Shewanella putrefaciens , a bacterium promising for bioremediation. PMID- 15539932 TI - Caveolin-1 in breast cancer. AB - Caveolin-1 is the principal structural protein of caveolae, sphingolipid and cholesterol-rich invaginations of the plasma membrane involved in vesicular trafficking and signal transduction. During caveolae-dependent signaling, caveolin-1 acts as a scaffold protein to sequester and organize multi-molecular signaling complexes involved in diverse cellular activities and, as such serves as a paradigm by which numerous disease processes may be affected by ablation or mutation of caveolin-1. The hypothesis that caveolin-1 conveys a tumor/transformation suppressor function in the mammary gland is derived from several independent lines of evidence accumulated by genetic, molecular and clinical approaches. The human caveolin-1 gene maps to a suspected tumor suppressor locus (D7S522/7q31.1) frequently deleted in human breast carcinomas. In addition, up to 16% of human breast carcinomas harbor a dominant-negative mutation, P132L, in the caveolin-1 gene. Caveolin-1 RNA and protein levels are also downregulated in human primary breast carcinomas and cell lines, with reintroduction of caveolin-1 in vitro sufficient to inhibit numerous tumorigenic properties, including anchorage independent growth and invasiveness. Most recently caveolin-1 knockout mice have provided breakthroughs in understanding the dynamic role of caveolin-1 in the pathogenesis of mammary epithelial cell hyperplasia, tumorigenesis and metastasis in a vivo setting. This review concentrates on recent advances implicating caveolin-1 in breast cancer pathogenesis, with emphasis on the signaling pathways regulated during these processes. PMID- 15539933 TI - Clipped wings: epigenetic inactivation of APC. PMID- 15539934 TI - NF-kappaB, IL-6 and myeloma cell growth: making the connection. PMID- 15539935 TI - Therapeutic targeting of prostate cancer. AB - An inaugural conference in Tucson Arizona on May 6-9, 2004 brought together more than 70 clinical and basic scientists to discuss recent research advances in understanding and targeting the progression of the human prostate cancer. The informal meeting was unique in that it provided the opportunity for discussion and interaction between these different groups of scientists whose paths rarely cross. The goal of the meeting was to develop new and novel approaches in understanding the human prostate cancer in order to uncover therapeutic targets. Faculty from six different cancer centers were represented including Memorial Sloan-Kettering Cancer Center (New York, NY); Arizona Cancer Center (Tucson, AZ); Fred Hutchinson Cancer Center (Seattle, WA); Chao Family Comprehensive Cancer Center (Irvine, CA); the Sydney Kimmel Cancer Center (San Diego, CA); Jonsson Comprehensive Cancer Center, University of California (Los Angeles, CA); and University of Massachusetts Memorial Cancer Center (Worcester, MA). Several important concepts emerged from this meeting as a result of the basic and clinical science interface. These concepts include: (1) Human prostate cancer has unique biological features as compared to other human epithelial malignancies; (2) Tumor plasticity is evident early in prostate cancer progression as evidenced by alterations in the extracellular matrix; (3) New therapeutic strategies should include the co-targeting of the stroma and prostate cancer; (4) Cell-cell and cell-ECM adhesion switching are reversible phenotypes evident early in human prostate tumor progression; (5) The discovery of molecular signatures including genomic or proteomic patterns for the discrimination of indolent versus aggressive disease is a potentially powerful tool and requires multifactorial approaches for success; and (6) New biomarkers and innovative tissue specific imaging modalities for human prostate cancer are being developed that may aid in a more accurate assessment of prostate cancer in patients. PMID- 15539936 TI - Genome-wide analysis of STAT target genes: elucidating the mechanism of STAT mediated oncogenesis. AB - Inappropriate activation of transcription factors is a common event in cancer. These transcription factors contribute to a malignant phenotype by regulating genes involved in cellular proliferation, survival, differentiation, angiogenesis, and invasion. An important goal remains identifying the targets of oncogenic transcription factors that execute these changes. STAT proteins are among the best-studied of these transcription factors, and are involved in oncogenesis both in vivo and in vitro. They thus represent an ideal model for understanding how transcription factors cause cancer through coordinated changes in gene expression. Recent studies have employed microarray-based expression analysis to comprehensively identify STAT target genes. Analysis of these targets can provide insight into mechanisms of neoplastic transformation, and may shed light on new strategies for targeted therapy. PMID- 15539937 TI - DNA hypomethylation is prevalent even in low-grade breast cancers. AB - Hypomethylation of some portions of the genome and hypermethylation of others are very frequent attributes of human cancer. We previously showed that cancer associated DNA hypomethylation often involves satellite 2 (Sat2), the main DNA component of the large juxtacentromeric (centromere-adjacent) heterochromatin of chromosome 1. In this study, we compared methylation of Sat2 and centromeric satellite DNA (Satalpha) as well as overall genomic methylation in 41 breast adenocarcinomas of known tumor grade and stage, 16 non-neoplastic breast tissues (mostly fibroadenomas), and a variety of normal somatic tissue controls. The cancers were significantly hypomethylated at Sat2 relative to the fibroadenomas or normal somatic tissues and at Satalpha relative to the normal somatic tissues. However, unlike Sat2, Satalpha did not display significant differences in methylation between the cancers and the non-neoplastic breast tissues. Therefore, hypomethylation at Sat2 is a much better marker of breast cancer than is Satalpha hypomethylation. There was a significant association of Sat2 hypomethylation with global DNA hypomethylation in the cancers but not with tumor grade, stage, axillary lymph node involvement, or hormone receptor status. Extensive cancer associated hypomethylation of juxtacentromeric satellite DNA and global DNA hypomethylation were common even in grade-1 or stage-1 carcinomas, which suggests that demethylation of the genome is an early event in breast carcinogenesis. PMID- 15539938 TI - Mechanism of inhibition of DNA methyltransferases by cytidine analogs in cancer therapy. AB - Hypermethylation of tumor suppressor genes caused by aberrant activity of DNA methyltransferases is an important mechanism that contributes to cancer. The reaction mechanism of DNA methyltransferases, which includes formation of a covalent intermediate between the enzyme and the target base, is the basis of the success of several anti-cancer drugs that are targeted against DNA methylation. These include 5-fluoro-2'-deoxycytidine, 5-aza-2'-deoxycytidine (Decitabine) and 2-H pyrimidinone-1-beta-D(2'-deoxyriboside) (Zebularine). This review provides an insight to how the chemistry of DNA methylation is involved in the performance of these drugs targeted against it. PMID- 15539939 TI - siRNAs: mechanism of RNA interference, in vivo and potential clinical applications. AB - Small interfering RNAs are currently the most widely used nucleic acid-based sequence-specific gene silencing molecules. These molecules mediate RNA interference--a natural post-transcriptional gene-silencing pathway. Given the high reliability and higher efficiency of small interfering RNA-mediated RNA interference, compared to earlier reverse genetic technologies, this is now the preferred technique in functional genomics. Furthermore, the exquisite specificity and exceptional gene-silencing potency of small interfering RNAs has resulted in intense research related to potential target-specific therapeutic applications of these molecules. This review will discuss the mechanism of RNA interference and applications of the pathway in molecular biology including functional genomics will be overviewed. The article will outline in vivo and potential clinical applications of small interfering RNA molecules. PMID- 15539940 TI - New strategy for efficient selection of dendritic cell-tumor hybrids and clonal heterogeneity of resulting hybrids. AB - Heterotypic hybrids created between dendritic cells (DC) and tumor cells represent an efficient approach for loading DC with tumor-associated antigens (TAA) and DC-tumor hybrid vaccines have shown promising outcomes in various preclinical and clinical studies. Conventional DC-tumor hybrid preparations, however, are unavoidably contaminated by DC-tumor aggregates and DC loaded with tumor cell debris. Here we describe a new strategy for selecting genuine DC-tumor hybrids. A HAT-sensitive/zeocin-resistant DC clone (XS106-7 Zeo) was fused with a GFP-transduced fibrosarcoma clone (S1509a-GFP) by polyethylene glycol and heterotypic hybrid clones were established by limiting dilution in the presence of HAT and zeocin. CD45 (DC origin) and GFP (tumor origin) were both expressed in 91% (51/56 clones) of the resulting clones, indicating high efficiency of our strategy. Marked heterogeneity was observed among the hybrid clones and only one clone exhibited characteristic features of DC (CD86 and I-A expression, dendritic morphology, T cell-stimulatory capacity and IL-1beta, IL-6 and TNFalpha production), suggesting that only small fractions of DC-tumor hybrids acquire and maintain the properties of parental DC. Finally, vaccination with this hybrid clone protected mice from subsequent growth of S1509a tumor cells, documenting the in vivo activity of DC-tumor hybrids in the complete absence of exogenous TAA. PMID- 15539941 TI - Raltitrexed increases tumorigenesis as a single agent yet exhibits anti-tumor synergy with 5-fluorouracil in ApcMin/+ mice. AB - The thymidylate synthase (TS) inhibitors raltitrexed (RTX) and 5-fluorouracil (FUra) have shown promising anti-tumor activity in preclinical and clinical settings for the treatment of colorectal cancer. Though the effects of these two agents have been reasonably well-characterized in cell lines, knowledge of their modes of action in vivo is limited. Here, we utilize the Apc(Min/+) mouse, an animal model of intestinal tumorigenesis, to study the effects of RTX treatment alone and in combination with FUra. Rather surprisingly, RTX monotherapy resulted in a dose dependent 4-10-fold increase in tumor number. The majority of these adenomas (74-95%) were rather small (i.e., less than 1 mm in diameter) and exhibited loss of heterozygosity at the Apc locus, suggesting an increase in mutational events leading to tumor development. RTX augmented BrdU-labeling of crypt epithelial cells, and retarded the movement of these cells along the crypt villus axis. Co-administration of FUra and RTX resulted in a significant reduction in tumor number compared to mice treated with either RTX or FUra alone (P < 0.0001). In addition, FUra abrogated the RTX-mediated increase in BrdU labeling. In all, the results show that RTX increases tumor burden in the Apc(Min/+) mouse, yet enhances the anti-tumor effect of FUra. This is the first illustration of in vivo synergy of RTX and FUra in a genetically predisposed animal model. Possible mechanisms underlying the current observations are discussed. PMID- 15539942 TI - The cancer story. PMID- 15539943 TI - Homeostatic regulation of base excision repair by a p53-induced phosphatase: linking stress response pathways with DNA repair proteins. AB - The p53 protein plays a central role in the integration of cellular stress responses. If the cell incurs DNA damage, p53-induced cell cycle arrest is accompanied by p53-facilitated DNA repair. In particular, p53 has been demonstrated to promote both nucleotide excision repair (NER) and base excision repair (BER). Once these repair processes are completed, p53 activity declines and the cell can reenter the cell cycle. A critical mediator of this p53 negative regulatory feedback loop is Mdm2, a p53 transcriptional target whose protein mediates p53 proteolytic degradation. Another such p53 transcriptional target that may function in a p53 negative regulation is the PPM1D phosphatase. PPM1D may inhibit p53 activity through inactivating dephosphorylation of the p38 MAP kinase. We have recently shown that PPM1D suppresses BER in part through dephosphorylation of a key BER effector, the nuclear isoform of uracil DNA glycosylase, or UNG2. This finding further links p53 signaling to DNA repair pathways and illustrates a mechanism by which activated DNA repair systems are returned to a deactivated, homeostatic state. PMID- 15539944 TI - BCCIP functions through p53 to regulate the expression of p21Waf1/Cip1. AB - The BCCIP protein is a BRCA2 and CDKN1A (p21(Waf1/Cip1)) Interacting Protein, which binds to a highly conserved domain of BRCA2, and a C-terminal domain of the CDK-inhibitor p21. We have previously reported that overexpression of BCCIP increases p21 mRNA and protein levels, and inhibits G(1) to S progression. In this report, we show that a partial shutdown of BCCIP expression by RNA interference reduces p21 levels and impairs G(1)/S checkpoint activation in response to ionizing radiation in HT1080 cells. We further show that the regulation of p21 expression by BCCIP is dependent on p53, and BCCIP regulates p53 transcription activity. These data provide a new mechanism by which BCCIP regulates p21 functions. PMID- 15539945 TI - Hepsin paradox reveals unexpected complexity of metastatic process. AB - The existing models of cancer progression assume that a linear sequence of genetic and epigenetic events occurs during this process. In this representation every new event (either loss of a tumor-suppressor, or activation of a proto oncogene) makes cells even more malignant. The result is a "super" cell that can form metastases at the distant sites. Metastatic cells are believed to carry all genetic and epigenetic characteristics that are necessary for metastasis formation. Recently, we have shown that cell-surface protease hepsin causes disorganization of the basement membrane and promotes prostate cancer progression and metastasis. In human prostate cancer hepsin is upregulated in the precancerous lesions and this upregulation is maintained in the primary tumors. Remarkably and completely unexpected for a metastasis-promoting gene, hepsin is expressed at low levels in metastatic lesions and the message is completely absent in metastasis-derived prostate cancer cell lines. These results demonstrate that genes that play an important role in metastatic process may exercise their role only at the specific fragments of cancer progression pathway (for example, during initial invasion and tissue disorganization in the primary organ) and may have no role in metastatic lesions. Future treatment of cancer patients may rely heavily on monitoring of tumor progression, as treatment efficient in attenuation of initial tumor progression may be inefficient or even adverse at the advance stages of disease. PMID- 15539946 TI - Hsp90 activation and cell cycle regulation. AB - The widely-expressed molecular chaperone heat shock protein 90 (Hsp90) regulates several important cellular processes via its' repertoire of 'client' proteins. Signal transduction pathways controlled by Hsp90 contribute to all major components of the malignant phenotype, so Hsp90 inhibitors are under investigation as anticancer agents. Since Hsp90 is also expressed at high levels in many normal tissues, it was unclear why Hsp90 inhibitors such as 17-allylamino geldanamycin (17-AAG) have selective antitumor activity in animals and are well tolerated clinically. Recent findings indicate that Hsp90 is largely latent in unstressed normal cells, but tumor Hsp90 becomes completely utilized during malignant progression, resulting in an activation-dependent conformational shift that radically increases 17-AAG binding affinity in cancer cells. In this article, the implications of this discovery are discussed, with particular reference to cell cycle regulation in normal and malignant cells, and the consequences of inducing cell cycle arrest with Hsp90 inhibitors. PMID- 15539947 TI - Flavopiridol, an inhibitor of transcription: implications, problems and solutions. AB - After a decade of exciting promises, the CDK inhibitor flavopiridol has quietly failed in most clinical trials. This review discusses that flavopiridol is a potent inhibitor of global transcription. This explains not only downregulation of numerous proteins, cell cycle arrest and apoptosis but also all pleiotropic and mysterious effects of flavopiridol. Yet, flavopiridol is not just a second actinomycin D. As an inhibitor of transcription with a unique mechanism of action, flavopiridol may have tremendous clinical potentials. This article reviews the molecular and cellular effects of flavopiridol as well as mechanisms of therapeutic and side effects, suggesting its novel clinical applications as a single agent and in drug combinations. PMID- 15539948 TI - ATR and ATM-dependent movement of BLM helicase during replication stress ensures optimal ATM activation and 53BP1 focus formation. AB - The BLM helicase, a deficiency that markedly increases cancer incidence in humans, is required for optimal repair during DNA replication. We show that BLM rapidly moves from PML nuclear bodies to damaged replication forks, returning to PML bodies several hours later, owing to activities of the DNA damage response kinases ATR and ATM, respectively. Immunofluorescence and cellular fractionation demonstrate that BLM partitions to different sub-cellular compartments after replication stress. Unexpectedly, fibroblasts lacking BLM were deficient in phospho-ATM (S-1981) and 53-binding protein-1 (53BP1), and these proteins failed to form foci following replication stress. Expression of a dominant p53 mutant or helicase-deficient BLM restored replication stress-induced 53BP1 foci, but only mutant p53 restored optimal ATM activation. Thus, optimal repair of damaged replication fork lesions likely requires both ATR and ATM. BLM recruits 53BP1 to these lesions independent of its helicase activity, and optimal activation of ATM requires both p53 and BLM helicase activities. PMID- 15539949 TI - Heat shock protein 70 promotes cancer cell viability by safeguarding lysosomal integrity. AB - The major heat-inducible Hsp70 is a potent survival protein that confers cytoprotection against numerous death-inducing stimuli and increases the tumorigenicity of rodent cells. The depletion of Hsp70 by adenovirus-mediated transfer of antisense cDNA induces caspase-independent death of tumorigenic cells while non-tumorigenic cells are unaffected, suggesting that Hsp70 has cancer specific function(s). We have recently demonstrated that the depletion of Hsp70 in cancer cells results in a cysteine cathepsin-dependent death, which is preceded by lysosomal destabilization and release of lysosomal constituents to the cytosol. In line with this, Hsp70 localizes to the membranes of lysosomes in human colon carcinoma cells and immortalized murine embryonic fibroblasts (MEFs) and prevents lysosomal membrane permeabilization and cell death induced by tumor necrosis factor (TNF), etoposide and H2O2. These findings identify Hsp70 as the first survival protein that functions by stabilizing the lysosomal membrane. PMID- 15539950 TI - Regulation of cell cycle in hematopoietic stem cells by the niche. AB - The quiescent state is thought to be an indispensable property for the maintenance of hematopoietic stem cells (HSCs). Interaction of HSCs with their particular microenvironments, known as the stem cell niches, is critical for cell cycle regulation of HSCs. Monitoring of the quiescence of HSCs using by a new stem cell marker, Side Population (SP), revealed that the cell cycle status of HSCs is dynamically controlled by the microenvironments. We have recently revealed a molecular mechanism in which cell cycle of HSCs is regulated by the niche. HSCs expressing the receptor tyrosine kinase Tie2 are adhere to osteoblasts (OBs) in the BM niche. The interaction of Tie2 and its ligand Angiopoietin-1 (Ang-1) leads to tight adhesion of HSCs to stromal cells, resulting in maintainance of long-term repopulating activity of HSCs. Thus, Tie2/Ang-1 signaling pathway plays a critical role in the maintenance of HSCs in a quiescent state in the BM niche. The understanding of cell cycle control in stem cells leads to development of new strategy for progress in regenerative medicine. PMID- 15539951 TI - Altered sumoylation of p63alpha contributes to the split-hand/foot malformation phenotype. AB - p63 mutations have been identified in several developmental abnormalities, including split-hand/foot malformation (SHFM). In this study, we demonstrate that the C-terminal domain of p63alpha associates with the E2 ubiquitin conjugating enzyme, Ubc9. A p63alpha mutation, Q634X, which naturally occurs in SHFM modulated the interaction of p63alpha with Ubc9 in yeast genetic assay. Furthermore, Ubc9 catalyzed the conjugation of p63alpha with small ubiquitin modifier-1 (SUMO-1), which covalently modified p63alpha in vitro and in vivo at two positions (K549E and K637E), each situated in a SUMO-1 modification consensus site (phiKXD/E). In addition, p63alpha mutations (K549E and K637E) abolished sumoylation of p63alpha, dramatically activated transactivation properties of TAp63alpha, and inhibited the dominant-negative effect of DeltaNp63alpha. These p63alpha mutations also affected the transcriptional regulation of gene targets involved in bone and tooth development (e.g., RUNX2 and MINT) and therefore might contribute to the molecular mechanisms underlying the SHFM phenotype. PMID- 15539952 TI - Epithelial-mesenchymal transition: NF-kappaB takes center stage. AB - Activation of the transcription factor NF-kappaB occurs in many human tumors, and studies have shown that NF-kappaB can promote cell proliferation and oncogenesis, possibly by protecting cells from apoptosis. Little is known, however, about whether NF-kappaB is involved in tumor progression including epithelial mesenchymal transition (EMT), a central process governing both morphogenesis and carcinoma progression in multicellular organisms. In a combined in vitro/in vivo model of mammary carcinogenesis, NF-kappaB was essential both for the induction and maintenance of EMT and for in vivo metastasis. NF-kappaB and the signaling pathways that are involved in its activation should therefore receive attention as potential targets for the development of novel anti-metastatic cancer treatments. PMID- 15539953 TI - Multiple roles for cysteine cathepsins in cancer. AB - Cysteine cathepsins are a family of proteases that have recently emerged as important players in cancer, and have variously been reported to be involved in apoptosis, angiogenesis, cell proliferation, and invasion. In normal cells, cysteine cathepsins are typically localized in lysosomes and other intracellular compartments, and are involved in protein degradation and processing. However, in certain tumors, cathepsins are translocated from their intracellular compartments to the cell surface, and can even be secreted. In addition, the expression and activity levels of some cysteine cathepsins are upregulated in human and mouse cancers. Understanding which cathepsins are critically involved, what their substrates are, and how they may be mediating these complex roles in cancer are important questions to address. We highlight recent results that begin to answer some of these questions, illustrating in particular the lessons from studying a mouse model of multistage carcinogenesis, which suggests distinctive roles for individual cysteine cathepsins in tumor progression. PMID- 15539954 TI - Pro- and anti-apoptotic effects of an inhibitor of chymotrypsin-like serine proteases. AB - The irreversible inhibitor of chymotrypsin-like serine proteases, N-tosyl-L phenylalanine chloromethylketone (TPCK), was shown to prevent internucleosomal DNA cleavage caused by inducers of apoptosis. The pro-apoptotic properties of TPCK have been studied less thoroughly. The aim of the present study was to investigate the pro- and anti-apoptotic activities of TPCK on HL-60 cells and compare them with the actions of the mitochondrial electron transport inhibitor antimycin A (AMA). The results showed that TPCK alone caused activation of cell cycle checkpoints, mitochondrial cytochrome c release, caspase-3 activation, and chromatin condensation. Caspase-8 was not required for cytochrome c release but was crucial to caspase-3 activation. TPCK synergistically enhanced AMA-induced cytochrome c release and caspase-3 activation while completely blocking AMA induced internucleosomal DNA fragmentation for at least 8 hours. Rather than blocking AMA-induced DNA fragmentation, the general serine protease inhibitor 4 (2-aminoethyl)-benzenesulphonyl fluoride (AEBSF) actually enhanced it. The pro apoptotic effect of TPCK may be due to activation of cell cycle checkpoints via inhibition of the proteasome. The apoptotic pathways activated by TPCK and AMA probably converge at the level of the mitochondria. The mode by which TPCK prevents internucleosomal DNA fragmentation is probably not through serine protease inhibition. PMID- 15539955 TI - Inhibition of mutant EGF receptors by gefitinib: targeting an Achilles' heel of lung cancer. AB - Recent reports revealed a strong correlation between the presence of somatic activating mutations in the kinase domain of the EGF receptor in some human lung cancers and a striking clinical response to the selective EGF receptor inhibitor, Gefitinib. These oncogenic receptors exhibit altered signaling properties such that they selectively activate downstream survival pathways on which tumor cells have become dependent. Thus, tumor cells are effectively killed by Gefitinib treatment. In addition, the catalytic function of the mutant receptors exhibits increased sensitivity to Gefitinib, raising the possibility that two distinct consequences of Gefitinib action account for its clinical efficacy. PMID- 15539956 TI - p53 prevents the accumulation of double-strand DNA breaks at stalled-replication forks induced by UV in human cells. AB - To investigate the mechanism by which UV irradiation causes S-phase-dependent chromosome aberrations and thereby genomic instability, we have developed an assay to study the DNA structure of replication forks (RFs) in UV-irradiated mammalian cells, using pulse-field gel electrophoresis for the DNA analysis. We demonstrate that replication stalling at UV-induced pyrimidine dimers results in the formation of single-strand DNA (ssDNA) regions and incomplete RF structures. In normal and in nucleotide-excision-repair (NER)-defective xeroderma pimentosum (XP) cells, stalling at dimers is rapid and prolonged and recovery depends on dimer repair or bypass. By contrast, XP variant (XPV) cells, defective in replication of a UV-damaged template due to mutation of bypass-polymerase epsilon, fail to arrest at dimers, resulting in a much higher frequency of ssDNA regions in the stalled RFs. We show that the stability of UV-arrested RFs depends directly on functional p53, and indirectly on NER and pol eta. In p53-deficient cells, the stalled sites give rise to double-strand DNA breaks (DSBs), at a frequency inversely correlated with repair capacity of the cell. In normal cells only a fraction of the stalled sites give rise to DSBs, while in XPASV, XPDSV and also XPVSV, all the sites do. XPVSV cells, although repair proficient, accumulate almost double the number of DSBs, suggesting that a high frequency of ssDNA regions in UV-arrested forks cause RF instability. These replication-associated DSBs do not accumulate in p53-proficient human cells. We propose that a major mechanism by which p53 maintains genome stability is the prevention of DSB accumulation at long-lived ssDNA regions in stalled-replication forks. PMID- 15539957 TI - WNT and cyclooxygenase-2 cross-talk accelerates adenoma growth. AB - Both Wnt and cyclooxygenase (COX-2) pathways are activated in most sporadic and familial colorectal cancers, especially in those with chromosomal instability. We have recently shown that a common target of both signaling pathways, the peroxisome proliferator-activated receptor (PPAR)-delta, is involved in intestinal adenoma growth. Activation of this receptor by synthetic agonist (GW501516) or COX-2-derived prostaglandin E2 (PGE2) accelerates intestinal adenoma growth in Apc(Min) mice. Moreover, these effects are lost in Apc(Min) mice lacking PPARdelta. These findings implicate PPARdelta as a focal point of cross-talk between the Wnt and prostaglandin signaling pathways. Based on this work it looks as if PPARdelta agonists currently in development for treatment of dyslipidemias and obesity may increase the risk of tumor formation in humans. By contrast, antagonists of PPARdelta may provide a novel approach for prevention and treatment of colorectal cancer. PMID- 15539958 TI - Chk1 is essential for tumor cell viability following activation of the replication checkpoint. AB - Chk1 (checkpoint kinase 1) is an evolutionarily conserved serine/threonine kinase involved in DNA damage responses. Originally identified as a kinase regulating the G2/M transition checkpoint, its role has broadened to include the S-phase checkpoint response and essential functions in early embryonic development. In this manuscript we investigated the potential of chemo-sensitization via ablation of Chk1 in cells treated with anti-metabolite cancer drugs, hydroxyurea (HU) and cytosine arabinoside (ara-C). Exposure to these replication interfering drugs in cells carrying Chk1 targeted siRNA provoked markedly increased rates of apoptosis. Although cell death was accompanied by an increase in p53 and activation of Chk2, the increased susceptibility to apoptosis was not dependent on p53 or Chk2. Additionally, we found that cells with reduced Chk1 expression displayed increased gamma-H2A.X expression, a marker for damaged DNA, and phosphorylated 32kDa subunit of replication protein A (RPA). Thus, Chk1 may play an essential role in maintaining DNA integrity during the replication block. Significantly, normal cells such as WS1 did not exhibit increased DNA damage or subsequent increases in apoptosis following replication stress, in the absence of Chk1. Thus, the essential role Chk1 plays in maintaining viability during the replication block in cancer cell lines can be exploited to sensitize cancer cells when abrogation of Chk1 is combined with DNA anti-metabolite chemotherapeutic drugs. Taken together, these data suggest that inhibition of Chk1 in combination with DNA anti-metabolite chemotherapy is a viable therapeutic strategy. PMID- 15539959 TI - Phosphorylation of Cdc25C by pp90Rsk contributes to a G2 cell cycle arrest in Xenopus cycling egg extracts. AB - In unfertilized Xenopus eggs, the p42 mitogen activated protein kinase (p42MAPK) pathway is known to maintain cell cycle arrest at metaphase of meiosis II. However, constitutive activation of p42MAPK in post-meiotic, cycling Xenopus egg extracts can lead to either a G2 or M-phase arrest of the cell cycle, depending on the timing of p42MAPK activation. Here, we examined the molecular mechanism by which activation of the p42MAPK pathway during interphase leads to cell cycle arrest in G2. When either a recombinant wild type Cdc25C(WT) or a mutated form of Cdc25C, in which serine 287 was replaced by an alanine (S287A), was added to cycling egg extracts, S287A accelerated entry into M-phase. Furthermore, the addition of S287A overcame the G2 arrest caused by p42MAPK, driving the extract into M-phase. p90Rsk a kinase that is the target of p42MAPK, was phosphorylated and activated (pp90Rsk) in the G2-arrested egg extracts, and was able to phosphorylate WT but not S287A in vitro. 14-3-3 proteins were associated with endogenous Cdc25C in G2-arrested extracts. Cdc25C(WT) that had been phosphorylated by pp90(Rsk) bound 14-3-3zeta, whereas S287A could not. These data suggest that the link between the p42MAPK signaling pathway and Cdc25C involves the activation of pp90Rsk and its phosphorylation of Cdc25C at S287, causing the binding of 14-3-3 proteins. We propose that the binding of 14-3-3 proteins to pp90Rsk phosphorylated-Cdc25C results in a G2 arrest in a manner similar to the cell cycle delays induced by differentiation signals that occur later in embryonic development. PMID- 15539960 TI - Low-density lipoprotein lowering--a race to the bottom. PMID- 15539961 TI - Atorvastatin reduces remnant lipoproteins and small, dense low-density lipoproteins regardless of the baseline lipid pattern. AB - Elevated plasma levels of remnant lipoproteins and small, dense low-density lipoprotein (LDL) particles increase the risk of atherosclerosis. This prospective, placebo-controlled, crossover trial evaluated the effect of atorvastatin on various lipid parameters including remnant lipoproteins and small, dense-LDL cholesterol levels. Forty-five subjects were enrolled in the study. These subjects fell into three distinct lipid patterns: atherogenic dyslipidemia, isolated hypercholesterolemia, and mixed dyslipidemia. Regardless of the baseline lipid profile, atorvastatin (10 mg q x d) reduced levels of remnant lipoproteins by 25%, LDL-cholesterol by 27%, and the three LDL subfractions by 23%-28% (p<0.0001 for all). Combining all patients, atorvastatin did not significantly alter the overall LDL subfraction pattern; however, in the isolated hypercholesterolemia group, the proportion of LDL present as the small, dense fraction increased by 23% (p=0.01) with treatment, whereas it did not change significantly in the other two groups. Overall, atorvastatin reduced triglycerides by 18% and apolipoprotein-B100 by 23% and increased high-density lipoproteins by 6.2% (p<0.001 all). Since atorvastatin is known to reduce the risk for coronary heart disease events and these data suggest that it does not appear to alter the LDL subfraction pattern, it is unclear whether or not the latter is an important risk predictor independent of LDL-cholesterol concentrations. Increased attention should be paid to absolute concentrations of LDL subfraction cholesterol, which may be a more sensitive indicator of coronary heart disease risk than total LDL or an LDL pattern. PMID- 15539962 TI - Subclinical calcified atherosclerosis in men and its association with a family history of premature coronary heart disease in first- and second-degree relatives. AB - The authors examined the associations between coronary artery calcification and a family history of premature coronary heart disease in either first-degree (immediate family) or second-degree (grandparents, aunts, uncles) relatives in 1619 asymptomatic healthy men ages 40-50 years. The prevalence of any coronary artery calcification was 19.3% in participants (n=1102) with no family history, 26.6% in those with a first-degree family history (n=203; 12.5%), 26.5% in those with a second-degree family history (n=215, 13.3%), and 30.3% with both (n=99, 6.1%, p=0.003). After controlling for the Framingham risk score, body mass index, and ethnicity, all categories of family history were significant predictors of coronary artery calcification. The odds ratios for coronary artery calcification associated with a first- (1.49; p=0.026) and second-degree (1.41; p=0.049) family history of coronary heart disease were similar. Clinical coronary risk assessments should broadly include an assessment of premature coronary heart disease in both first- and second-degree relatives. PMID- 15539963 TI - Intensive lifestyle modification: impact on cardiovascular disease risk factors in subjects with and without clinical cardiovascular disease. AB - Intensive lifestyle modification programs are intended to stabilize or promote regression of coronary artery disease; however, clinical response is often nonuniform, complicating appropriate utilization of resources and prediction of outcome. This study assessed physiological and psychological benefits to 72 persons participating in a prospective, nonrandomized, four-component lifestyle change program and compared response between patients with clinical cardiovascular disease (CVD) and patients with elevated risk factors for CVD but without clinical manifestations of disease. Subjects entering the program due to elevated risk factor levels alone demonstrated equal or greater benefit, in terms of improvement in primary CVD risk factors and reduction in measures of coronary disease risk developed in the Framingham Heart Study, than those with clinical CVD. These findings suggest that intensive lifestyle change programs may be important for primary prevention in individuals at increased risk of CVD. PMID- 15539965 TI - The effects of niacin on lipoprotein subclass distribution. AB - Dyslipidemia is a heterogeneous metabolic condition; high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very-low-density lipoprotein represent families of lipoprotein particles that differ in size and composition and vary in atherogenicity. Lipoprotein subclasses containing apolipoprotein B promote atherosclerosis, of which the most atherogenic appear to be the small, dense LDL and large very-low-density lipoprotein subclasses, while the large HDL2 subclass, which transports esterified cholesterol from the periphery to the liver, is considered the more cardioprotective. Niacin has long been known to improve concentrations of all major lipids and lipoproteins, but it also has consistently favorable effects on subclass distribution. A MEDLINE search was conducted for clinical studies reporting the effects of niacin on lipoprotein subclasses. The niacin-associated elevations in HDL cholesterol likely stem from differential drug effects on subclasses, producing favorable changes in levels of HDL2 and apolipoprotein A-I. Niacin has more moderate LDL cholesterol-lowering efficacy, but this change is associated with an increase in LDL particle size and a shift from small LDL to the less atherogenic, large LDL subclasses. In addition, it also tends to decrease concentrations of the larger very-low-density lipoprotein subclasses. Niacin confers diverse benefits with respect to both the quantity and quality of lipid and lipoprotein particles. PMID- 15539966 TI - Physical activity and markers of inflammation and thrombosis related to coronary heart disease. AB - Epidemiologic studies have provided strong evidence of the beneficial effects of physical activity in patients with coronary heart disease; however, the mechanisms for the cardioprotective effect of physical activity are not fully understood. The more favorable changes in coronary risk profiles of physically active individuals and the favorable changes observed on several of the established coronary risk factors in exercise intervention studies must play a partial role in that protection. In more recent years, inflammatory and thrombotic factors have been implicated in the development and progression of atherosclerosis. Physical activity has been associated with lower levels of several inflammatory markers; however, the effect of physical activity on coagulation markers remains controversial. The authors summarize the studies on the effect of physical activity on several inflammatory and thrombotic variables. The importance of physical activity in the prevention of cardiovascular disease through modification of the inflammatory and thrombogenic processes is also addressed. PMID- 15539964 TI - Rosuvastatin alone or with extended-release niacin: a new therapeutic option for patients with combined hyperlipidemia. AB - Combination therapy with a statin and niacin may provide optimal therapy for patients with combined hyperlipidemia and low levels of high-density lipoprotein (HDL) cholesterol. The authors assessed the efficacy and safety of rosuvastatin monotherapy, extended-release (ER) niacin monotherapy, or rosuvastatin and ER niacin combined therapy in patients with atherogenic dyslipidemia. In a 24-week, open-label, multicenter trial, men and women aged > or =18 years with fasting levels of total cholesterol > or =200 mg/dL, HDL cholesterol > or =45 mg/dL, triglycerides 200-800 mg/dL, and apolipoprotein B > or =110 mg/dL were randomly assigned to one of four treatment groups: rosuvastatin 10-40 mg, ER niacin 0.5-2 g, rosuvastatin 40 mg plus ER niacin 0.5-1 g, or rosuvastatin 10 mg plus ER niacin 0.5-2 g. Daily doses of rosuvastatin 40 mg monotherapy reduced low-density lipoprotein (LDL) cholesterol and non-HDL cholesterol levels significantly more than did either ER niacin 2 g monotherapy or rosuvastatin 10 mg combined with ER niacin 2 g. Addition of ER niacin 1 g to rosuvastatin 40 mg did not further reduce total or non-HDL cholesterol. Triglyceride reductions were similar among the four treatment groups. ER niacin mono- and combined therapy produced significantly greater rises in HDL cholesterol and apolipoprotein A-1 than did rosuvastatin monotherapy. Rosuvastatin monotherapy was better tolerated than ER niacin taken either alone or with rosuvastatin. In this study, rosuvastatin very effectively improved the three major lipoprotein-lipid abnormalities of combined hyperlipidemia. PMID- 15539969 TI - Inflammation in atherothrombosis: how to use high-sensitivity C-reactive protein (hsCRP) in clinical practice. AB - Inflammation is now recognized as a critical contributor to the atherothrombotic process, and measurement of the inflammatory biomarker high-sensitivity C reactive protein (hsCRP) is a proven method to better predict cardiovascular risk and target therapeutic interventions. Following recommendations from the Centers for Disease Control and the American Heart Association, many physicians now routinely evaluate hsCRP along with cholesterol levels as a part of global risk prediction. Levels of hsCRP <1, 1-3, and >3 mg/L correspond to lower, moderate, and higher risk of cardiovascular events at all levels of the Framingham Risk Score and at all levels of metabolic syndrome. Interventions that lower hsCRP include diet, exercise, smoking cessation, statin therapy, and improved glycemic control. In addition to primary prevention, hsCRP is an important prognostic marker in acute coronary syndromes, following angioplasty, and in the long-term management of post-infarction patients. This article provides a clinically oriented overview of appropriate settings in which to measure hsCRP, how to interpret results, and how interventions to reduce vascular risk can be targeted on the basis of hsCRP findings. PMID- 15539970 TI - Acute heart failure: patient management of a growing epidemic. AB - Acute heart failure is a rapidly growing clinical problem in the United States. There are few randomized clinical trials to guide treatment; however, important observational data are now emerging from the Acute Decompensated Heart Failure National Registry regarding the demographics and treatment of these patients. Management consists largely of identification and treatment of precipitating factors, correction of comorbid conditions, and IV diuretics and vasodilators. Nitroprusside is a valuable treatment, but its use is usually restricted to patients in the intensive care unit who are undergoing hemodynamic monitoring. Nesiritide is being increasingly employed. Specialized strategies such as dialysis, continuous venous-venous hemodialysis, extra-corporeal membrane oxygenator, left ventricular assist device, and heart transplantation are employed in a small subset of patients. Although recovery is the rule, the in hospital mortality for acute heart failure is high and the readmission rate is very high. Prevention of acute heart failure by avoiding factors known to precipitate decompensation remains the most cost-effective strategy. PMID- 15539971 TI - Pharmacologic advances in long-term heart failure management. AB - Advances in the pharmacologic management of patients with heart failure during the last several decades have been mainly guided by the results of well conducted, randomized controlled clinical trials (RCTs). As a consequence of these RCTs, we now know that angiotensin-converting enzyme inhibitors and beta blockers should be concomitantly administered to patients who tolerate these therapies with the expectation that reductions in cardiovascular deaths and hospital admissions for management of heart failure will be accomplished. New therapeutic modalities must now be proven to add incremental clinical benefits to these establish therapies. This challenge is currently being addressed with aldosterone blocking agents as well as angiotensin receptor blockers. This article discusses the major RCTs in this field and places the new trials in perspective. PMID- 15539972 TI - Acute coronary syndromes: pathogenesis, acute diagnosis with risk stratification, and treatment. AB - Acute ischemic chest pain at rest consistent with unstable angina or non-ST elevation myocardial infarction is a common problem that may cause death or recurrent myocardial infarction within 30 days unless identified and risk stratified acutely. The latter may be done within 15 minutes by the history, physical exam, and electrocardiogram, and is aided by the measurement of troponin T/I. According to the Agency for Health Care Policy and Research guidelines, low risk patients can be discharged home and rechecked within 72 hours. Intermediate risk patients with no ST-segment changes with continuous monitoring and no elevation of troponin should undergo exercise stress testing by electrocardiogram (or nuclear or echocardiographic evaluation if electrocardiogram is non analyzable). Patients with a negative stress test are low risk (no death or myocardial infarction at 30 days or 6 months) and can be discharged home. Patients with a positive test or who are at high risk according to the Agency for Health Care Policy and Research guidelines should undergo acute invasive testing for possible revascularization. Aspirin and low molecular weight heparin or unfractionated heparin, along with anti-ischemia therapy, is indicated in intermediate- or high-risk patients. The addition of clopidogrel is indicated in these patients, except in those who are potential candidates for coronary artery bypass graft. Platelet glycoprotein IIb/IIIa inhibitors are indicated in high risk patients likely to undergo percutaneous coronary intervention, should be started early if recurrent ischemia occurs, but are not indicated in lower-risk patients who do not require percutaneous coronary intervention. Intensive secondary prevention should be started before dismissal. PMID- 15539973 TI - Stroke prevention in atrial fibrillation: anticoagulants and antithrombotics. AB - Atrial fibrillation and heart failure are growing epidemics in the developed world and often coexist. From clinical trials, warfarin is highly effective in reducing stroke in patients with atrial fibrillation. Equally important is the fact that in spite of well-designed trials, translation of the results of the data from the trials into clinical practice has been less than optimal. One of the reasons is that warfarin is a difficult drug to use. Thus there has been a concerted effort to develop an alternative to warfarin. Ximelagatran and Dabigatran, both direct thrombin inhibitors, are the furthest along in clinical development. Ximelagatran, while highly effective as an anticoagulant and safe with regard to bleeding, has been associated with liver function abnormalities; the importance of which needs resolution. Dabigatran is much earlier in development and is currently of unproven value. It is highly likely that alternatives to warfarin for stroke prevention will be available in the future and will likely result in a higher utilization rate of anticoagulants in patients with atrial fibrillation. PMID- 15539974 TI - Obstructive hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM) is a primary disease of cardiac muscle characterized by a thickening of the left ventricular wall and often predominantly affecting the interventricular septum. This paper presents a case study of a 53-year-old female with a dynamic and obstructive form of HCM. The study includes a case presentation, clinical findings, investigations, and management. Patient findings were obtained before and after alcohol septal ablation, a novel interventional therapy. The article reviews the various forms of HCM and describes the currently available treatment modalities for obstructive HCM. Further research will be necessary to determine the comparative efficacy between past, present, and future therapies. PMID- 15539975 TI - The relationship of medication regimen to hospital readmissions for older adults with heart failure. AB - The purpose of this part of a longitudinal study was to examine whether medication therapy for older adults with heart failure predicted days to readmission post-hospital discharge. Using a prospective, predictive design, a convenience sample included 127 older adults with heart failure who had been recently discharged from two hospitals in northeastern Ohio. One hundred five patients were prescribed diuretics, 49 angiotensin-converting enzyme inhibitors, 23 b blockers, and 47 digoxin. There were no significant differences between readmitted and non-readmitted patients with regard to the use of the specific classes of cardiac medications. None of the specific classes of cardiac medications predicted the number of days between the initial hospital discharge and readmission 3 months later. The use of a small, non-probability sample and exclusion of variables limit the results of the study. Effective case management with teaching about heart failure must address changes involved with heart failure and the use of medication therapy. More research is needed about treatment protocols in various regions of the United States. PMID- 15539976 TI - Using motivational interviewing to reduce diabetes risk. AB - With the growing prevalence of diabetes worldwide, controlling modifiable risk factors for diabetes is essential to preventing complications and disease progression. Recent research strongly supports targeting at-risk prediabetes clients through therapeutic lifestyle change. Many behavior change techniques and strategies are known to be successful, yet are seldom implemented in today's health care arena. Nurses are in an excellent position to serve as change agents to assist at-risk prediabetes clients in making necessary lifestyle changes. Motivational interviewing has been shown to be effective in counseling clients toward behavior change. The major principles of motivational interviewing will be described and motivational interviewing techniques will be demonstrated using a scenario with a prediabetes client. PMID- 15539977 TI - The effects of obesity on the cardiopulmonary system: implications for critical care nursing. AB - Obesity has risen at epidemic rates over the last 20 years. This increase crosses all ages, genders, racial/ethnic groups, and income and educational levels. A variety of genetic, environmental, physiological, psychological, and sociocultural factors influence the development of obesity. Increased morbidity and mortality are associated with obesity. Most health-related problems and the increased risk of death in obesity are due to cardiopulmonary compromise. This paper addresses the physiologic effects of obesity on the cardiopulmonary system and the impact obesity has on the critically ill patient. Suggestions for alterations in cardiovascular and pulmonary assessment techniques are discussed. Nursing practice interventions are presented for maximizing cardiopulmonary function and preventing complications. The advocacy role of the nurse is addressed as a critical component in working to develop a culture of understanding and acceptance for the obese patient. PMID- 15539978 TI - International perspectives on CV nursing: cardiovascular nursing in Ireland. PMID- 15539979 TI - New guidelines for low-density lipoprotein levels from the National Cholesterol Education Program (NCEP): a 2004 update. PMID- 15539980 TI - Improving outcomes: management of hyperglycemia. PMID- 15539981 TI - Ximelagatran-an oral direct thrombin inhibitor. PMID- 15539982 TI - What is pacemaker tracking? PMID- 15539986 TI - Assessing wounds accurately. PMID- 15539990 TI - Exactly what is a tax home and do I need one? PMID- 15539992 TI - Winter travel assignments appeal to RN, ski bum. PMID- 15539994 TI - Traveling with technology: PDAs to go. PMID- 15539995 TI - Quick tips for first-time travelers. PMID- 15539996 TI - Top 10 reasons to love travel nursing. PMID- 15539997 TI - Walk away from foot pain. PMID- 15540037 TI - Surveying sedation and analgesia practice in the pediatric intensive care unit: discomforting data raise further questions. PMID- 15540038 TI - Serum troponin detection suggests myocardial injury in pediatric sepsis...what next? PMID- 15540039 TI - Stress in parents of children in the pediatric intensive care unit. PMID- 15540041 TI - Where will your influence end? PMID- 15540040 TI - Tezosentan, another selective selection for pulmonary hypertension, or not? PMID- 15540042 TI - The importance of CNS specialization. PMID- 15540043 TI - CNS entrepreneurs: innovators in patient care. PMID- 15540044 TI - Does faculty practice validate the NP educator role? PMID- 15540045 TI - What will national provider identifiers mean for you? PMID- 15540046 TI - Quiet revolutions: the CRNA scope of practice. PMID- 15540047 TI - The art of using visual aids. PMID- 15540048 TI - Nurse-midwifery: yesterday, today, and tomorrow. PMID- 15540049 TI - Break through language barriers. PMID- 15540050 TI - Choosing malpractice insurance. PMID- 15540059 TI - Just the facts. PMID- 15540061 TI - Criteria for selecting an online educational offering. PMID- 15540062 TI - Ethics and genetics: implications for CNS practice. PMID- 15540063 TI - New hope for patients with pulmonary hypertension: endothelin receptor antagonists: update for the clinical nurse specialist. PMID- 15540064 TI - Expert to novice: a journey from administrator to entrepreneur. PMID- 15540065 TI - ABC codes: a new opportunity to capture CNS contributions to US healthcare. AB - A specialized set of insurance-related codes for integrative healthcare will increase the national influence and patient-specific effectiveness of clinical nurse specialists (CNSs) and others in the US healthcare system because virtually every participant in the healthcare system uses codes for some critical purpose. Inadequate codes prevent decision makers from having essential health-related information at their fingertips. ABC codes improve business processes and health industry efficiencies in the same manner that universal product codes (UPCs/bar codes) improved the retail operations and the retail industry. The codes are essential to research, management, and commerce. In healthcare, the combination of ABC codes with older medical and dental code sets allows many essential business processes (such as forecasting, practice management, insurance billing, claims processing, and third-party reimbursement) to be fully automated. The codes also help digitize information to simplify data collection, analysis, and reporting. This improves the quality of data and accelerates the speed with which conclusions can be drawn from those data about what works in US healthcare and why. By offering CNSs more complete, accurate, and precise information on best practices, ABC codes help ensure that CNSs and others can generate better outcomes in addressing health-related challenges faced by patients/clients, nurses, and nursing practices, and the nation's health-promoting organizations and systems. PMID- 15540067 TI - Development and implementation of an online CNS program. AB - PURPOSE/OBJECTIVES: Online educational programs are rapidly increasing in all fields. This article describes a private university's online clinical nurse specialist graduate program that included supervised clinical practicum experiences. METHODS: The program was designed using the 1998 National Association of Clinical Nurse Specialists (NACNS) Statement on Clinical Nurse Specialist Practice and Education as a curriculum guide. Issues that occur with online program development/design, implementation, and evaluation are described and solutions are presented. CONCLUSIONS: The online education program for clinical nurse specialists faced challenges; however, faculty and students agreed that this innovative program is a success. PMID- 15540068 TI - The Ottawa Model of Research Use: a guide to clinical innovation in the NICU. AB - PURPOSE/OBJECTIVES: To improve performance of a neonatal transport team by implementing a research-based family assessment instrument. Objectives included providing a structure for evaluating families and fostering the healthcare relationship. BACKGROUND/RATIONALE: Neonatal transports are associated with family crises. Transport teams require a comprehensive framework to accurately assess family responses to adversity and tools to guide their practice toward parental mastery of the event. Currently, there are no assessment tools that merge family nursing expertise with neonatal transport. DESCRIPTION OF THE PROJECT: A family assessment tool grounded in contemporary family nursing theory and research was developed by a clinical nurse specialist. The Ottawa Model of Research Use guided the process of piloting the innovation with members of a transport team. Focus groups, interviews, and surveys were conducted to create profiles of barriers and facilitators to research use by team members. Tailored research transfer strategies were enacted based on the profile results. OUTCOME: Formative evaluations demonstrated improvements in team members' perceptions of their knowledge, family centeredness, and ability to assess and intervene with families. The family assessment tool is currently being incorporated into Clinical Practice Guidelines for Transport and thus will be considered standard care. CONCLUSION: Use of a family assessment tool is an effective way of appraising families and addressing suffering. The Ottawa Model of Research Use provided a framework for implementing the clinical innovation. IMPLICATIONS FOR NURSING PRACTICE: A key role of the clinical nurse specialist is to influence nursing practice by fostering research use by practitioners. When developing and implementing a clinical innovation, input from end users and consumers is pivotal. Incorporating the innovation into a practice guideline provides a structure to imbed research evidence into practice. PMID- 15540069 TI - Dr Hildegard Peplau. PMID- 15540072 TI - Patient safety: it's not just carefulness, it's a culture. PMID- 15540073 TI - Triumph and tragedy: implementing a physician-focused program to impact length of stay, part II. PMID- 15540074 TI - Understanding the incomprehensible: a guide to the new Medicare prescription drug benefit for case managers. AB - Much of the health news over the last few months has centered on problems elderly patients encounter in obtaining and effectively using the prescription drug discount cards that became available on June 1 under the Medicare Prescription Drug, Improvement, and Modernization Act of 2003. This article focuses on the next prescription drug newsmaker, Medicare Part D, that will supercede the discount cards on January 1, 2006. There are many complex issues that case managers must evaluate when assisting beneficiaries with queries about "what to do." This article attempts to clarify the "incomprehensible" twists and turns of these issues and provides access to a "Medicare Drug Prescription Benefit Calculator" that may assist the beneficiary and case manager in decision making. Case managers need to understand that there are many opposing viewpoints on this benefit, and it promises to become the subject of a major national debate. For this reason, substantial changes may occur prior to the launch of Part D. If you think discount drug cards are confusing, "you ain't seen nothin' yet!" PMID- 15540075 TI - Patient safety systems for case management. AB - Patient safety is an important concern of many healthcare stakeholders, including patients, providers, employers, health plans, and insurers. In 2003, URAC conducted a qualitative study to examine the role of utilization management programs as part of a systemic approach to promoting patient safety. Many of URAC's findings are applicable to case management as well. URAC found that most medical management companies address patient safety indirectly, as part of a global quality management program. Even so, the study identified a number of strengths that could be deployed by medical management organizations to more systematically promote patient safety. For example, case management organizations use decision support tools to assist frontline practitioners in conducting assessments and tracking interventions. Such systems could be programmed to flag safety concerns and guide interventions. The study also found that commonly used case management assessment protocols could be adapted to routinely assess for high priority safety concerns. URAC concluded that each stage of the medical management process offers opportunities for data collection and clinical interface to promote patient safety. Case managers have key positions interfacing between patients, providers, and the healthcare system. Development of safety indicators and training of staff are key elements needed to effectuate a safety program in medical management. Case management leaders in each organization and at the national level should make patient safety a priority, and define the processes for implementing safety systems wherever case management is practiced. PMID- 15540077 TI - Integrating services for optimal proactive care. AB - While case management, disease management and health advocacy each approach the patient relationship from a different angle, there are opportunities to integrate the three to achieve better results and move past finding only the sickest 1-3% in order to produce cost stabilization for employers. This article looks at how and why we should add survey-based predictive tools and coaching to the current case management and disease management mix to achieve substantial cost reduction and successful intervention. PMID- 15540078 TI - The neuropsychologist's role in concussion management. PMID- 15540079 TI - Meperidine and the management of pain: what you need to know. PMID- 15540080 TI - Building a successful private care management company. PMID- 15540112 TI - Viable, apoptotic and necrotic monocytes expose phosphatidylserine: cooperative binding of the ligand Annexin V to dying but not viable cells and implications for PS-dependent clearance. PMID- 15540113 TI - XIAP-deficiency leads to delayed lobuloalveolar development in the mammary gland. PMID- 15540114 TI - Sensitization of osteosarcoma cells to death receptor-mediated apoptosis by HDAC inhibitors through downregulation of cellular FLIP. AB - Fas-mediated apoptosis plays an important role in elimination of tumor cells in vivo, but some tumor-derived cells are resistant to this mechanism. Here, we show that treatment with the histone deacetylase (HDAC) inhibitor FR901228 renders Fas resistant osteosarcoma cell lines sensitive to Fas-mediated apoptosis by downregulating expression of cellular FLIP (cellular FLICE-inhibitory protein), an inhibitor of Fas-mediated activation of caspase-8. Moreover, sensitization to Fas-mediated apoptosis was also induced in Fas-resistant osteosarcoma cells by suppressing FLIP expression using FLIP-specific RNA interference. HDAC inhibitors including FR901228 were shown to induce downregulation of cellular FLIP through inhibiting generation of FLIP mRNA, rather than stimulating degradation at either protein or mRNA level, and the inhibition was independent of de novo protein synthesis. These results clearly indicate that some tumor cells exhibit a phenotype resistant to death receptor-mediated apoptosis by expressing cellular FLIP, and that HDAC inhibitors sensitize such resistant tumor cells by directly downregulating cellular FLIP mRNA. PMID- 15540115 TI - Efficient infection and persistence of a herpesvirus saimiri-based gene delivery vector into human tumor xenografts and multicellular spheroid cultures. AB - Herpesvirus saimiri (HVS) has the ability to infect a variety of human cell lines and establish a persistent infection by virtue of episomal maintenance. Moreover, the viral episome provides sustained expression of a heterologous transgene. HVS based vectors can also persist for a long term in tumor xenografts generated from HVS-infected human carcinoma cell lines. The viral episome remains latent within the xenograft allowing long-term transgene expression. These properties, in addition to its ability to incorporate large amounts of heterologous DNA, make HVS an attractive potential gene delivery vector. Here we report on the further evaluation of such HVS-based vectors. We demonstrate for the first time that HVS can efficiently infect solid tumor xenografts derived from a variety of human carcinoma cells via direct intratumoral injections. Furthermore, HVS can efficiently infect spheroid cultures, a three-dimensional cell culture system that closely resembles a tumor. Upon infection of both the tumor xenografts and spheroid cultures, HVS-based vectors can establish a persistent episomal infection within the tumor xenograft allowing expression of a heterologous transgene. These results suggest that HVS-based vectors may be suitable for cancer gene therapy applications. PMID- 15540116 TI - Crystal structure of yeast V-ATPase subunit C reveals its stator function. AB - Vacuolar H(+)-ATPase (V-ATPase) has a crucial role in the vacuolar system of eukaryotic cells. It provides most of the energy required for transport systems that utilize the proton-motive force that is generated by ATP hydrolysis. Some, but not all, of the V-ATPase subunits are homologous to those of F-ATPase and the nonhomologous subunits determine the unique features of V-ATPase. We determined the crystal structure of V-ATPase subunit C (Vma5p), which does not show any homology with F-ATPase subunits, at 1.75 A resolution. The structural features suggest that subunit C functions as a flexible stator that holds together the catalytic and membrane sectors of the enzyme. A second crystal form that was solved at 2.9 A resolution supports the flexible nature of subunit C. These structures provide a framework for exploring the unique mechanistic features of V ATPases. PMID- 15540117 TI - ARF6-dependent interaction of the TWIK1 K+ channel with EFA6, a GDP/GTP exchange factor for ARF6. AB - TWIK1 belongs to a family of K(+) channels involved in neuronal excitability and cell volume regulation. Its tissue distribution suggests a role in epithelial potassium transport. Here we show that TWIK1 is expressed in a subapical compartment in renal proximal tubules and in polarized MDCK cells. In nonpolarized cells, this compartment corresponds to pericentriolar recycling endosomes. We identified EFA6, an exchange factor for the small G protein ADP ribosylation factor 6 (ARF6), as a protein binding to TWIK1. EFA6 interacts with TWIK1 only when it is bound to ARF6. Because ARF6 modulates endocytosis at the apical surface of epithelial cells, the ARF6/EFA6/TWIK1 association is probably important for channel internalization and recycling. PMID- 15540118 TI - Multiple gastrointestinal stromal tumors in type I neurofibromatosis: a pathologic and molecular study. AB - Multiple gastrointestinal stromal tumors typically occur in familial form associated with KIT receptor tyrosine kinase or platelet-derived growth factor receptor-alpha (PDGFRA) germline mutations, but may also develop in the setting of type 1 neurofibromatosis. The molecular abnormalities of gastrointestinal stromal tumors arising in neurofibromatosis have not been extensively studied. We identified three patients with type 1 neuro-fibromatosis and multiple small intestinal stromal tumors. Immunostains for CD117, CD34, desmin, actins, S-100 protein, and keratins were performed on all of the tumors. DNA was extracted from representative paraffin blocks from separate tumor nodules in each case and subjected to a nested polymerase chain reaction, using primers for KIT exons 9, 11, 13, and 17 and PDGFRA exons 12 and 18, followed by direct sequencing. The mean patient age was 56 years (range: 37-86 years, male/female ratio: 2/1). One patient had three tumors, one had five, and one had greater than 10 tumor nodules, all of which demonstrated histologic features characteristic of gastrointestinal stromal tumors and stained strongly for CD117 and CD34. One patient died of disease at 35 months, one was disease free at 12 months and one was lost to follow-up. DNA extracts from 10 gastrointestinal stromal tumors (three from each of two patients and four from one patient) were subjected to polymerase chain reactions and assessed for mutations. All of the tumors were wild type for KIT exons 9, 13, and 17 and PDGFRA exons 12 and 18. Three tumors from one patient had identical point mutations in KIT exon 11, whereas the other tumors were wild type at this locus. We conclude that, although most patients with type 1 neurofibromatosis and gastrointestinal stromal tumors do not have KIT or PDGFRA mutations, KIT germline mutations might be implicated in the pathogenesis of gastrointestinal stromal tumors in some patients. PMID- 15540120 TI - Experimental and quantum chemical studies of structure and reaction mechanisms of dioxouranium(VI) complexes in solution. AB - This perspective article describes the combination of experimental data and quantum chemical methods for the determination of structure and reaction mechanisms of uranyl(vi) complexes in aqueous solution. The first part assesses the accuracy of the chemical and thermodynamic properties of solvated uranyl(vi) complexes as obtained by various quantum chemical methods. The second part discusses structure determination, mechanisms for ligand exchange and the lability of coordinated water molecules for various uranyl(vi) complexes using a combination of NMR and quantum chemical data. PMID- 15540119 TI - Gains of 13q are correlated with a poor prognosis in liposarcoma. AB - Liposarcomas are a phenotypical heterogeneous group of tumors divided into four main subtypes: well-differentiated, dedifferentiated, myxoid/round cell, and pleomorphic. The aim of this study was to compare DNA sequence copy number changes of these subtypes as investigated by comparative genomic hybridization in 36 patients. Comparative genomic hybridization revealed genomic imbalances in tumors of 27 patients (mean 5.6 imbalances per tumor). The most frequent gains were within single regions of 1q, 12q, and 13q. We found a significant correlation of poor overall survival and gain of 13q21 (P=0.0221), 13q22 (P=0.0341), 13q31 (P=0.0410), and 13q32 (P=0.0074). The univariate Cox regression analysis revealed an increased risk of tumor-related death for patients whose liposarcomas possess with gains of 13q21 and 13q32 simultaneously (P=0.010; RR=7.1; 95% CI 1.6-31.7). Furthermore, 12 high-level amplifications were found in tumors of seven patients. In four cases 12q14-q15 and in two cases 13q32-q33 were amplified. We identified in different liposarcoma subtypes characteristic genomic changes: Gains and high-level amplifications of 12q occurred in all 11 investigated well-differentiated liposarcomas, and these changes were often present simultaneously with gains of 1q (mean 5.5 changes). In the two dedifferentiated liposarcomas, gains of 1q in both liposarcomas, and a high-level amplification of 13q were striking. Only eight of the 17 patients with myxoid/round cell liposarcomas showed changes in DNA copy number (mean 3.4 imbalances). In four of these eight cases gains of 13q occurred. The six pleomorphic liposarcomas possessed the most frequent genomic imbalances (mean number 16.3) of all liposarcoma subtypes investigated. These imbalances were in almost all chromosomal regions detected predominantly as over-representations of chromosomes 1, 5p, 13q, and 22q. Summarizing, all subtypes but well differentiated liposarcomas showed gains of 13q, which were associated with a poor prognosis. PMID- 15540121 TI - First transition-metal complexes of S=P(NHBu t)3. AB - The first transition-metal (Rh(I), Mo(VI), Ni(II)) complexes of S[double bond, length as m-dash]P(NHBu(t))(3) have been synthesized via metathetical reactions of mono-lithiated and [Rh(CO)(2)Cl](2), (Bu(t)N)(2)MoCl(2)(dme) and NiBr(2)(dme). Surprisingly in the molecular structure of the Ni(II)-complex both hard-soft (N,S) and hard-hard (N,N[prime or minute]) chelation modes of are realized. PMID- 15540122 TI - The effects of hydrofluoric acid addition on the hydrothermal synthesis of templated uranium sulfates. AB - The effect of employing hydrofluoric acid as a mineraliser in the formation of organically templated uranium sulfate materials has been studied. Variable amounts of HF((aq)) were added to a series of reaction gels in which all other reactant concentrations were invariant, resulting in the formation of three different phases, depending upon the fluoride concentration. Two of these phases are novel; [N(2)C(4)H(14)][UO(2)(H(2)O)(SO(4))(2)] is a new templated uranium sulfate, containing anionic [UO(2)(H(2)O)(SO(4))(2)](2-) chains that hydrogen bond to one-another forming pseudo-layers, and [N(2)C(4)H(14)][UO(2)F(SO(4))](2) is the first example of a templated uranium sulfate fluoride, which consists of uranium fluoride chains linked by sulfate groups to form [UO(2)F(SO(4))](-) layers. The role of F(-) in these reactions is two-fold; it acts as a mineraliser when present in small concentrations, while it is incorporated into the reaction product when present in larger mole fractions. Both of the new materials have been characterised using a range of physical techniques including single crystal X-ray structure analysis. PMID- 15540123 TI - Reduced and excited states of the intermediates (alpha-diimine)(C5R5)Rh in hydride transfer catalysis schemes: EPR and resonance Raman spectroscopy, and comparative DFT calculations of Co, Rh and Ir analogues. AB - The electronic structures of the highly air-sensitive intermediates (N[caret]N) (C(5)Me(5))Rh, (N[caret]N = 2,2'-bipyridine (bpy), 2,2'-bipyrimidine (bpym), 2,2' bipyrazine (bpz) and 3,3'-bipyridazine (bpdz)) of hydride transfer catalysis schemes were studied through resonance Raman (rR) spectroscopy and through EPR of the reduced forms [(N[caret]N) (C(5)Me(5))Rh](.-). The rR results are compatible with a predominant MLCT character of the lowest excited states [ (N[caret]N) (C(5)Me(5))Rh]*, and the EPR spectra of the reduced states reveal the presence of anion radical ligands, (N[caret]N) (.-), coordinated by unusually electron rich rhodium(i) centres. The experimental results, including the assignments of electronic transitions, are supported by DFT calculations for the model compounds [(N[caret]N)(C(5)H(5))Rh](o)/(.-), (N[caret]N) = bpy or bpym. The calculations confirm a significant but not complete mixing of metal and ligand orbitals in the lowest unoccupied MO which still retains about 3/4 pi* (N[caret]N) character. DFT calculations on (bpy)(C(5)H(5))M and [(bpy)(C(5)H(5))ClM](+), M = Co, Rh, Ir, agree with the experimental results such as the differences between the homologues, especially the different LUMO characters of the precursor cations in the case of Co-->d(M)) and Rh or Ir (-->pi*(bpy)). PMID- 15540124 TI - Sulfur, tin and gold derivatives of 1-(2'-pyridyl)-ortho-carborane, 1-R-2-X-1,2 C2B10H10 (R = 2'-pyridyl, X = SH, SnMe3 or AuPPh3). AB - Reaction of the lithium salt of 1-(2'-pyridyl)-ortho-carborane, Li[1-R-1,2 C(2)B(10)H(10)](R = 2'-NC(5)H(4)), with sulfur, followed by hydrolysis, gave the mercapto-o-carborane, 1-R-2-SH-1,2-C(2)B(10)H(10) which forms chiral crystals containing helical chains of molecules linked by intermolecular S-H...N hydrogen bonds. The cage C(1)-C(2) and exo C(2)-S bond lengths (1.730(3) and 1.775(2)[Angstrom], respectively) are indicative of exo S=C pi bonding. The tin derivative 1-R-2-SnMe(3)-1,2-C(2)B(10)H(10), prepared from Li[1-R-1,2 C(2)B(10)H(10)] and Me(3)SnCl, crystallises with no significant intermolecular interactions. The pyridyl group lies in the C(1)-C(2)-Sn plane, oriented to minimise the NSn distance (2.861(3)[Angstrom]). The tin environment is distorted trigonal bipyramidal with axial N and Me. The gold derivative 1-R-2-AuPPh(3)-1,2 C(2)B(10)H(10), prepared from Li[1-R-1,2-C(2)B(10)H(10)] and AuCl(PPh(3)), reveals no NAu interaction in its crystal structure. PMID- 15540125 TI - Host-guest assembly of ligand systems for metal ion complexation. Synergistic solvent extraction of copper(II) and silver(I) by 1,4,8,11-tetrabenzyl-1,4,8,11 tetraazacyclodecane in combination with carboxylic acids. AB - Host-guest formation between 1,4,8,11-tetrabenzyl-1,4,8,11-tetraazacyclodecane (1) and lipophilic organic carboxylic acids in chloroform has been investigated and the effect of such ligand assembly on the solvent extraction of copper(II) and silver(I) has been probed. NMR titration experiments in the absence of a metal ion confirm the formation of weak 1:1 and 1:2 (macrocycle:carboxylic acid) assemblies in CDCl(3) between 1 and palmitic (hexadecanoic) acid or 4-tert butylbenzoic acid while difunctional salicylic acid showed a 1 [ratio] 2 interaction that is somewhat stronger. The interaction between the former two acids and the tetra-N-benzylated macrocycle is significantly less than that reported previously for its non-substituted parent, cyclam; a result that likely reflects the presence of the less-basic, more sterically hindered tertiary nitrogens in 1 relative to the secondary nitrogens present in cyclam. Carboxylic acid-containing assemblies of this type have been used as extractants in a series of solvent extraction (water/chloroform) experiments. From both previous observations as well as from entropy considerations, it was anticipated that the use of a host-guest assembly of the above type for metal-ion complexation might contribute to enhanced metal ion binding (and concomitant enhanced metal ion extraction). Such behaviour is postulated to arise from the components of the coordination sphere being, at least in part, assembled for complex formation. In accord with this, the use of the ligand assembly involving palmitic acid/macrocycle 1 was found to lead to enhanced (synergistic) extraction of copper(II) at a metal ion concentration of 10(-3) mol dm(-3) while, for silver(I), synergism was somewhat marginal at this concentration but was clearly apparent under related conditions when the silver concentration was reduced to 10(-4) mol dm(-3). Similar behaviour towards silver was also observed when 4-tert butylbenzoic acid was substituted for palmitic acid, while the use of salicylic acid resulted in enhanced (synergistic) extraction at both metal ion concentrations. PMID- 15540126 TI - Kinetic and mechanistic study on the reaction of alkylcobaloximes with azoles. AB - The kinetics of axial water substitution by azoles (pyrazole and 1,2,4-triazole) in three different cobaloximes, viz.trans-[Co(Hdmg)(2)(R)H(2)O] where Hdmg = dimethylglyoximate, R = PhCH(2), Et and CF(3)CH(2), were studied as a function of azole concentration, temperature and pressure in aqueous solution. The second order rate constants for the substitution of water in trans-[Co(Hdmg)(2)(R)H(2)O] for R = Et at pH 6.0, 25 degrees C and I= 0.1 M (NaClO(4)), were found to be 1309 and 1200 M(-1) s(-1) for pyrazole (Pz) and 1,2,4-triazole (Tz), respectively, and those obtained for R = PhCH(2) were found to be 755 and 691 M(-1) s(-1), respectively. The second order rate constants in the case of R = CF(3)CH(2) were found to be 0.358 and 0.348 M(-1) s(-1) for Pz and Tz, respectively. The relative order of reactivity for the different alkyls being Et > PhCH(2) > CF(3)CH(2). The activation parameters (DeltaH([not equal]), DeltaS([not equal]) and DeltaV([not equal])) obtained for these reactions were found to be in the range of 65-87 kJ mol(-1), 24-47 J mol(-1) K(-1) and 2.5-7.7 cm(3) mol(-1), respectively. These data suggest that an I(d) substitution mechanism operates where the azoles participate in the transition state. PMID- 15540127 TI - Synthesis and structure of Zn7(mu4-O)2(OAc)10(Pz)2(OAc = acetate; Pz = pyrazine). AB - Reaction of Zn(OAc)(2).2H(2)O with pyrazine in refluxing ethanol gives the unusual heptanuclear complex Zn7(mu4-O)(2)(OAc)(10)(Pz)(2) (1) (OAc = acetate, Pz = pyrazine) in 46% yield. A single-crystal X-ray diffraction study of revealed a central Zn(7) core in which two pseudo-tetrahedral Zn(4) units are joined at a common vertex. The two pyrazine molecules are bound as terminal (eta1) ligands. PMID- 15540128 TI - Dinuclear and mononuclear manganese(IV)-radical complexes and their catalytic catecholase activity. AB - Seven o-aminophenol ligands based on aniline and m-phenylenediamine, which act as mono- and di-nucleating non-innocent ligands, respectively, together with their seven Mn(IV)-complexes, 1-7 are described. One of them, 1, is dinuclear and 2-7 are mononuclear, in which the ligands are coordinated in their oxidized o iminobenzosemiquinone radical forms. The crystal structures of 1 and 3 were determined by X-ray diffraction and the electronic structures were established by various physical methods including EPR and variable-temperature (2-290 K) susceptibility measurements. Electrochemical measurements (CV and SQW) indicate primarily ligand-centered redox processes. The Mn(IV)-radical complexes, 1-7, catalyze the oxidation of 3,5-di-tert-butylcatechol with molecular oxygen as the sole oxidant to afford 3,5-di-tert-butylquinone quantitatively under mild conditions to mimic the function of the copper-containing enzyme catechol oxidase. An "on-off" mechanism of the radicals without redox participation of the metal center is proposed for the catalytic oxidation process. Complex 1 is found to be a good catalyst for oxidative C-C coupling of hindered phenols to diphenoquinones. PMID- 15540129 TI - Synthesis and characterization of mono- and binuclear platinum complexes containing terminal and bridging diynyldiphenylphosphine ligands. AB - A series of mononuclear platinum complexes containing diynyldiphenylphosphine ligands [cis-Pt(C(6)F(5))(2)(PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CR)L](n)(n= 0, L = tht, R = Ph 2a, Bu(t)2b; L = PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CR, 4a, 4b; n=-1, L = CN(-), 3a, 3b) has been synthesized and the X-ray crystal structures of 4a and 4b have been determined. In order to compare the eta2-bonding capability of the inner and outer alkyne units, the reactivity of towards [cis-Pt(C(6)F(5))(2)(thf)(2)] or [Pt(eta2) C(2)H(4))(PPh(3))(2)] has been examined. Complexes coordinate the fragment "cis Pt(C(6)F(5))(2)" using the inner alkynyl fragment and the sulfur of the tht ligand giving rise the binuclear derivatives [(C(6)F(5))(2)Pt(mu-tht)(mu 1kappaP:2eta2-C(alpha),C(beta)-PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CR)Pt(C(6)F(5))(2)](R = Ph 5a, Bu(t)5b). The phenyldiynylphosphine complexes 2a, 3a and 4a react with [Pt(eta2)-C(2)H(4))(PPh(3))(2)] to give the mixed valence Pt(II)-Pt(0) complexes [((C(6)F(5))(2)LPt(mu-1kappaP:2eta2)-C(5),C(6) PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CPh))Pt(PPh(3))(2)](n)(L = tht 6a, CN 8a and PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CPh 9a) in which the Pt(0) fragment is eta2-complexed by the outer fragment. Complex 6a isomerizes in solution to a final complex [((C(6)F(5))(2)(tht)Pt(mu-1kappaP:2eta2) C(alpha),C(beta)-PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CPh))Pt(PPh(3))(2)]7a having the Pt(0) fragment coordinated to the inner alkyne function. In contrast, the tert-butyldiynylphosphine complexes 2b and 3b coordinate the Pt(0) unit through the phosphorus substituted inner acetylenic entity yielding 7b and 8b. By using 4a and 2 equiv. of [Pt(eta2)-C(2)H(4))(PPh(3))(2)] as precursors, the synthesis of the trinuclear complex [cis-((C(6)F(5))(2)Pt(mu-1kappaP:2eta2) C(5),C(6)-PPh(2)C[triple bond]CC(6)H(4)C[triple bond]CPh)(2))(Pt(PPh(3))(2))(2)]10a, bearing two Pt(0)(PPh(3))(2)eta2) coordinated to the outer alkyne functions is achieved. The structure of 7a has been confirmed by single-crystal X-ray diffraction. PMID- 15540130 TI - Di- and tri-alkylphosphine adducts of S-donor palladacycles as catalysts in the Suzuki coupling of aryl chlorides. AB - The reaction of tricyclohexylphosphine with the S-based palladacycle [(Pd(mu OAc)(kappa2-S,C-C(6)H(4)CH(2)SMe))(2)] gives several products, regardless of stoichiometry, one of which, [Pd(kappa1-OAc)(eta1-C(6)H(4)CH(2)SMe)(PCy3)2], has been characterised crystallographically. Despite this, catalysts formed in situ from di- and tri-alkylphosphines and [(Pd(mu-OAc)(kappa2-S,C-C(6)H(4)CH(2)SMe))2] show excellent activity in the Suzuki coupling of a range of deactivated, non activated and activated aryl chloride substrates. PMID- 15540131 TI - Equilibrium, kinetic and HPLC study of the reactions between platinum(II) complexes and DNA constituents in the presence and absence of glutathione. AB - The complex formation equilibria of [Pt(SMC)(H(2)O)(2)](+) and [Pt(terpy)H(2)O](2+), where SMC =S-methyl-L-cysteine and terpy = 2,2':6',2" terpyridine, with some biologically relevant ligands such as inosine (INO), inosine-5'-monophosphate (5'-IMP), guanosine-5'-monophosphate (5'-GMP) and glutathione (GSH) were studied. The stoichiometry and stability constants of the complexes formed are reported, and the concentration distribution of the various complex species have been evaluated as a function of pH. Also the kinetics and mechanism of the complex formation reactions were studied as a function of nucleophile concentration and temperature. For the complex [Pt(SMC)(H(2)O)(2)](+), two consecutive reaction steps, which both depend on the nucleophile concentration, were observed under all conditions. The negative entropies of activation support an associative complex formation mechanism. Reaction of guanosine-5'-monophosphate (5'-GMP) with Pt(II) complexes was carried out in the presence and absence of glutathione (GSH) at neutral pH. The rate constants clearly showed a kinetic preference toward GSH at neutral pH. The reactions were also monitored by HPLC. However, only a small amount of coordinated 5'-GMP was detected in the HPLC trace. The products were isolated and characterized by MALDI-TOF mass spectrometry. PMID- 15540132 TI - An enormous vibrational motion: the gas-phase structure of dimethyl bis(methoxyethynyl) germanium. AB - The structure of dimethyl-bis(methoxyethynyl) germanium has been determined in the gas phase by electron diffraction utilising flexible restraints from quantum chemical calculations. Theoretical methods (B3LYP/6-311+G* and MP2/6-311+G*) predict a low barrier to rotation of the methoxy groups in the molecule in addition to low-frequency vibrations of the long ethynyl chains. In the equilibrium structure the Ge-C[triple bond]C angles of the two methoxyethynyl fragments in the molecule are computed to deviate by up to 4 degrees from the linear arrangement. As a consequence of low-frequency large-amplitude vibrational motion the experimental structure of these fragments without applying vibrational corrections deviates considerably from linearity, while the structure corrected for vibrational effects using the harmonic approximation and taking into account a non-linear transformation between internal and Cartesian coordinates (r(h1)) shows closer agreement with theory. The main experimental structural parameters of dimethyl-bis(methoxyethynyl) germanium (r(h1)) are: r(Ge-C)(mean), 192.5(1) pm; DeltaGeC =r(Ge-C(methyl))-r(Ge-C(ethynyl)), 4.5(5) pm, r(C[triple bond]C)(mean), 122.8(2) pm; r(C-O)(mean), 138.9(3) pm; DeltaCO =r(C(methyl)-O) r(C(ethynyl)-O), 14.5(2) pm, r(C-H)(mean), 109.1(4) pm; [angle](X-C-H)(mean)(X = Ge,O), 109(1) degree; [angle]C(ethynyl)-Ge-C(ethynyl), 108.1(4) degree; [angle]C(methyl)-Ge-C(methyl), 113.4(5) degree; [angle]Ge-C[triple bond]C, 163(1) degree; [angle]C[triple bond]C-O, 176(2) degree; [angle]C-O-C, 115.2(6) degree; methoxy group torsion, tau, 36(9) degree from the position in which the C-O bond eclipses the further Ge-C(ethynyl) bond. PMID- 15540133 TI - Transition metal complexes of the chelating phosphine borane ligand Ph2PCH2Ph2P.BH3. AB - Chromium and ruthenium complexes of the chelating phosphine borane H(3)B.dppm are reported. Addition of H(3)B.dppm to [Cr(CO)(4)(nbd)](nbd = norbornadiene) affords [Cr(CO)(4)(eta1-H(3)B.dppm)] in which the borane is linked to the metal through a single B-H-Cr interaction. Addition of H(3)B.dppm to [CpRu(PR(3))(NCMe)(2)](+)(Cp =eta5)-C(5)H(5)) results in [CpRu(PR(3))(eta1-H(3)B.dppm)][PF(6)](R = Me, OMe) which also show a single B-H-Ru interaction. Reaction with [CpRu(NCMe)(3)](+) only resulted in a mixture of products. In contrast, with [Cp*Ru(NCMe)(3)](+)(Cp*=eta5)-C(5)Me(5)) a single product is isolated in high yield: [Cp*Ru(eta2-H(3)B.dppm)][PF(6)]. This complex shows two B-H-Ru interactions. Reaction with L = PMe(3) or CO breaks one of these and the complexes [Cp*Ru(L)(eta1-H(3)B.dppm)][PF(6)] are formed in good yield. With L = MeCN an equilibrium is established between [Cp*Ru(eta2-H(3)B.dppm)][PF(6)] and the acetonitrile adduct. [Cp*Ru (eta2-H(3)B.dppm)][PF(6)] can be considered as being "operationally unsaturated", effectively acting as a source of 16-electron [Cp*Ru (eta1-H(3)B.dppm)][PF(6)]. All the new compounds (apart from the CO and MeCN adducts) have been characterised by X-ray crystallography. The solid-state structure of H(3)B.dppm is also reported. PMID- 15540134 TI - Synthesis and structural characterization of two novel heterometallic clusters: [Rh4Pt2(CO)11(dppm)2] and [Ru2Rh2Pt2(CO)12(dppm)2]. AB - Two novel heterometallic octahedral clusters [Rh(4)Pt(2)(CO)(11)(dppm)(2)](1) and [Ru(2)Rh(2)Pt(2)(CO)(12)(dppm)(2)](2) were synthesized by the reaction of [Rh(2)Pt(2)(CO)(6)(dppm)(2)] with [Rh(6)(CO)(14)(NCMe)(2)] and Ru(3)(CO)(12), respectively. Solid state structures of 1 and 2 have been established by a single crystal X-ray diffraction study. Two dppm ligands in 1 are bonded to one platinum and three rhodium atoms, which form an equatorial plane of the Rh(4)Pt(2) octahedron. Two rhodium and two platinum atoms bound to the diphosphine ligands in 2 are nonplanar to give an octahedral C2 symmetric Ru(2)Rh(2)Pt(2)(dppm)2 framework. The (31)P NMR investigation of and (1D, (31)P COSY, (31)P-[(103)Rh] HMQC) and simulation of 1D spectral patterns showed that in both clusters the structures of the M(6)(PP)(2) fragments found in the solid state are maintained in solution. PMID- 15540135 TI - Alkalicyamelurates, M3[C6N7O3].xH2O, M = Li, Na, K, Rb, Cs: UV-luminescent and thermally very stable ionic tri-s-triazine derivatives. AB - Cyamelurates are salts of cyameluric acid, a derivative of tri-s-triazine (1,3,4,6,7,9-hexaazacyclo[3.3.3]azine or s-heptazine). These compounds are thermally very stable and possess interesting structural and optical properties. Only very few tri-s-triazine derivatives have been reported in the literature. The water-soluble alkali cyamelurates were extensively characterized using NMR, FTIR, Raman, UV, luminescence spectroscopy and elemental analysis. In addition, the single crystal X-ray structure analyses of the four hydrates of lithium, sodium, potassium and rubidium cyamelurates (Li(3)[C(6)N(7)O(3)].6H(2)O; Na(3)[C(6)N(7)O(3)].4.5H(2)O; K(3)[C(6)N(7)O(3)].3H(2)O; Rb(3)[C(6)N(7)O(3)].3H(2)O) are presented. Thermogravimetric analysis shows that the dehydrated salts start to decompose at temperatures above 500 degrees C. The thermal stability does not depend on the cations which is in contrast to the analogous s-triazine salts, i.e. the alkali cyanurates M(3)[C(3)N(3)O(3)]. The photoluminescence spectra indicate a very strong solid state UV-emission with maxima between 280 and 400 nm. PMID- 15540136 TI - Synthesis and characterization of half-sandwich N-heterocyclic carbene complexes of cobalt and rhodium. AB - A series of novel half-sandwich M(I) and M(III) complexes (M = Co, Rh) bearing the N-heterocyclic carbene ligand 1,3-dimesitylimidazol-2-ylidene (IMes) have been prepared and characterized. Thus, (eta5-C(5)R(5))M(IMes)(C(2)H(4))(M = Co, Rh; R = H, Me) were obtained from the corresponding bis(ethene) complexes (eta5 C(5)R(5))M(C(2)H(4))(2), except for CpRh(IMes)(C(2)H(4)) which was prepared via the novel 16-electron Rh(I) compound Rh(IMes)(C(2)H(4))(2)Cl. The carbonyl compounds (eta5-C(5)R(5))Co(IMes)(CO)(R = H, Me) were synthesized by thermal CO substitution of (eta5-C(5)R(5))Co(CO)(2). A diamagnetic, apparently 16-electron Co(III) compound [CpCo(IMes)I](+)[I(3)(-)] was obtained from CpCo(IMes)(CO) and I(2). Finally, Co(III) and Rh(III) complexes CpCo(IMes)Me(2) and Cp*Rh(IMes)Me(2) were prepared by methylation of [CpCo(IMes)I](+)[I(3)(-)], and ligand exchange at Cp*Rh(Me(2)SO)Me(2), respectively. The molecular structures of CpCo(IMes)(CO), CpRh(IMes)(C(2)H(4)), Cp*Rh(IMes)(C(2)H(4)), and Cp*Rh(IMes)Me(2) were determined by single crystal X-ray diffraction. Steric and electronic factors imposed by the strongly donating and sterically demanding IMes ligand are discussed on the basis of X-ray crystallographic, NMR, and IR spectroscopic analyses. Very poor correlations are found between values for (1)J(Rh-C(carbene)) and dRh-C(carbene) data for Rh(i) N,N-heterocyclic carbene complexes including literature data and this work. PMID- 15540137 TI - New types of the flexible self-assembled metal-organic coordination polymers constructed by aliphatic dicarboxylates and rigid bidentate nitrogen ligands. AB - The reaction of metal ions, flexible aliphatic dicarboxylates and rigid bidentate linear ligands under mild conditions in water afford four novel metal-organic coordination polymers, [Cd(mu-mal)(mu-pyz)(0.5)(H(2)O)](n) 1 (mal = malonate dianion, pyz = pyrazine), [Cd(2)(mu-suc)(2)(mu-pyz)(H(2)O)(2)](n) 2 (suc = succinate dianion), and ([M(mu-bipy)(H(2)O)4][suc].4H(2)O)(n)(M = Co, 3, M = Zn, 4, bipy = 4,4'-bipyridine). The molecular structures of 1-4 have been established by single-crystal X-ray crystallography. 1 is a 3D network being composed of layers of octahedrally coordinated Cd atoms bridged by malonate anions in syn anti configurations within the layers and pyz molecules between layers. Unlike that in 1, each Cd atom in 2 displays uncommon pentagonal-bipyramidal geometry to form 2D infinite grid sheets with square grid dimensions of ca. 7.936 x 7.936 [Angstrom]. Both 3 and 4 exhibit 1D linear -M-bipy-M-bipy- chain polymers, and these chains were packed as ...ABCABC... layered structures. The bridging succinate ligands in 2 adopt the syn-anti mode with a torsion angle of 60.8(7) degrees, while the solvated succinate ligands in 3 and 4 adopt the anti-anti mode with a torsion angle of 180.0 degrees. To our knowledge, compound 2 represents the first example of flexible self-assembled succinate-pyrazine mixed bridging ligand coordination network. 3 and 4 are the first two cases of succinate-bipy polymers with non-coordinated succinate. The magnetic behavior for 3 was studied in the temperature range of 5-300 K. The result indicates the occurrence of a weak antiferromagnetic coupling between the cobalt(II) ions. PMID- 15540139 TI - The evolution of virulence when parasites cause host castration and gigantism. AB - It has been suggested that the harm parasites cause to their hosts is an unavoidable consequence of parasite reproduction with costs not only for the host but also for the parasite. Castrating parasites are thought to minimize their costs by reducing host fecundity, which may minimize the chances of killing both host and parasite prematurely. We conducted a series of experiments to understand the evolution of virulence of a castrating bacterium in the planktonic crustacean Daphnia magna. By manipulating food levels during the infection of D. magna with the bacterium Pasteuria ramosa, we showed that both antagonists are resource limited and that a negative correlation between host and parasite reproduction exists, indicating resource competition among the antagonists. Pasteuria ramosa also induces enhanced growth of its hosts (gigantism), which we found to be negatively correlated with host fecundity but positively correlated with parasite reproduction. Because infected hosts never recovered from infections, we concluded that gigantism is beneficial only for the parasite. Hosts, however, have evolved counteradaptations. We showed that infected hosts have enhanced reproduction before castration. This shift to earlier reproduction increases overall host fecundity and compromises parasite reproduction. Finally, we showed that this resource conflict is subject to genetic variation among host and parasite genotypes within a population and is therefore likely to be an important force in the coevolution of virulence in this system. A verbal model is presented and suggests that the adaptive value of gigantism is to store host resources, which are liberated after parasitic castration for later use by the growing parasite. This hypothesis assumes that infections are long lasting, that is, that they have a high life expectancy. PMID- 15540140 TI - Do hosts and parasites coevolve? Empirical support from the Schistosoma system. AB - Coevolution between host and parasite is, in principle, a powerful determinant of the biology and genetics of infection and disease. However, coevolution is difficult to demonstrate rigorously in practice and therefore has rarely been observed empirically, particularly in animal-parasite systems. Research on host schistosome interactions has the potential for making an important contribution to the study of coevolution or reciprocal adaptation. This may be particularly pertinent because schistosomes represent an indirectly transmitted macroparasite, so often overlooked among both theoretical and empirical studies. Here we present ideas and experiments on host-schistosome interactions, in part reviewed from published work but focusing in particular on preliminary novel data from our ongoing studies of potential host-schistosome evolution and coevolution in the laboratory. The article is split into three main sections: we first focus on the evidence for evolution in the host, then in the parasite, before combining both to illustrate the gathering evidence of host-parasite coevolution in the snail schistosome system. In particular, we demonstrate that genetic architecture, variability, and selective pressures are present for the evolution of resistance and susceptibility, virulence, and infectivity to occur, the mechanisms allowing such polymorphisms to be maintained, and that hosts and parasites appear to have reciprocal effects on each other's phenotype and genotype. PMID- 15540141 TI - The rise and fall of a six-year coral-fungal epizootic. AB - Drivers of disease cycles are poorly understood in marine ecosystems in spite of increasing outbreaks. We monitored a newly emerged fungal epizootic (aspergillosis) affecting sea fan corals (Gorgonia ventalina L.) in the Florida Keys to evaluate causes of its rise and fall over 6 years. Since August 1997, aspergillosis has nearly eradicated large sea fans at some sites. However, sea fan densities have remained relatively constant due to episodic recruitment replacing large fans with small. Recruitment itself was affected by infection and occurred only when prevalence of disease was low. This impact on recruitment occurred because the largest, potentially most fecund colonies had the highest prevalence of disease, and the pathogen significantly suppressed reproduction of infected fans. Moreover, high mortality among adults resulted in a demographic shift to smaller colonies. The most dramatic impact of aspergillosis was the Keys wide loss of >50% of sea fan tissue from complete and partial mortality. Aspergillosis prevalence has declined steadily over the last 6 years, and we consider the following hypotheses for decline of the epizootic: change in environment, change in pathogen input, and increase in host resistance. We conclude that increasing host resistance is the most likely driver of the decline. However, a change in any of a number of factors, for example, recruitment of naive hosts, rate of pathogen input, or environmental conditions (water quality and temperature), is likely to promote reemergence of the epizootic. PMID- 15540142 TI - Host sex and local adaptation by parasites in a snail-trematode interaction. AB - One of the leading theories for the evolutionary stability of sex in eukaryotes relies on parasite-mediated selection against locally common host genotypes (the Red Queen hypothesis). As such, parasites would be expected to be better at infecting sympatric host populations than allopatric host populations. Here we examined all published and unpublished infection experiments on a snail-trematode system (Potamopyrgus antipodarum and Microphallus sp., respectively). A meta analysis demonstrated significant local adaptation by the parasite, and a variance components analysis showed that the variance due to the host-parasite interaction far exceeded the variance due to the main effects of host source and parasite source. The meta-analysis also indicated that asexual host populations were more resistant to allopatric sources of parasites than were (mostly) sexual host populations, but we found no significant differences among parasite populations in the strength of local adaptation. This result suggests that triploid asexual snails are more resistant to remote sources of parasites, but the parasite has, through coevolution, overcome the difference. Finally, we found that the degree of local adaptation did not depend on the genetic distance among host populations. Taken together, the results demonstrate that the parasites are adapted, on average, to infecting their local host populations and suggest that they may be a factor in selecting against common host genotypes in natural populations. PMID- 15540143 TI - Population dynamics of pathogens with multiple host species. AB - Pathogens that can infect multiple host species will have different dynamics than pathogens that are restricted to a single species of host. This article examines the conditions for establishment and long-term population dynamic behavior of pathogens that infect multiple species of hosts. The article attempts to address three major questions in this area: First, under which conditions will increases in the diversity of host species buffer infectious disease outbreaks, and under which conditions will host diversity amplify disease outbreaks? Second, under what conditions is it possible to control an infectious agent by focusing control against only one host species? Third, what role does host species diversity play in maintaining pathogen persistence? The answers to these questions supply some important general insights into the role that biodiversity plays in buffering humans and other species against new and emerging pathogens. PMID- 15540144 TI - Pathogen spillover in disease epidemics. AB - In field experiments manipulating generalist pathogens and host community composition, the presence of a highly susceptible reservoir species drove disease dynamics in multiple nonreservoir species, sometimes decreasing their abundance through apparent competition. The dynamics of generalist pathogens in multispecies host communities remain a major frontier for disease ecology. Of particular interest are how host community structure controls pathogen transmission and how disease spread feeds back to influence the host community. Pathogen spillover occurs when epidemics in a host population are driven not by transmission within that population but by transmission from a reservoir population. Here we review examples of spillover in pathogens infecting humans, domesticated animals, and crops, noting that most empirical evidence for spillover results from nonmanipulative, observational studies. We then present results from two field experiments utilizing an experimentally tractable model system of annual wild grasses and a generalist virus, the barley yellow dwarf virus. In these experiments, the presence of a highly susceptible reservoir species, Avena fatua (wild oats), greatly increased pathogen prevalence in several other species. This result demonstrates pathogen spillover and illustrates the crucial role of host community structure in controlling the dynamics of generalist pathogens. Further, pathogen spillover from A. fatua decreased the abundance of two other host species through pathogen-mediated apparent competition. Thus, our results provide experimental support for theoretical predictions of strong feedbacks between host community structure and generalist disease dynamics. PMID- 15540145 TI - Parasites and the evolutionary diversification of primate clades. AB - Coevolutionary interactions such as those between hosts and parasites have been regarded as an underlying cause of evolutionary diversification, but evidence from natural populations is limited. Among primates and other mammalian groups, measures of host diversification rates vary widely among lineages, but comparative studies have not yet identified a reliable explanation for this variation. In this study, we used a comprehensive data set of disease-causing organisms from free-living primates to illustrate how phylogenetic comparative methods can be used to examine mammalian lineage diversity in relation to parasite species richness. Our results provide evidence that the phylogenetic diversity of primate clades is correlated positively with the number of parasite species harbored by each host and that this pattern is largely independent of other host traits that have been shown to influence diversification rates and parasite species richness in primates. We investigated two possible mechanisms that could explain this association, namely that parasites themselves drive host evolutionary diversification through processes linked with sexual selection and that host shifts or host sharing increases parasite species richness among diverse primate clades. Neither parasite species richness nor host diversification is related to measures of sexual selection in primates. Further, we found only partial evidence that more rapidly diversifying host lineages produced increased opportunities for host sharing or host shifting by parasites through mechanisms involving species' geographic range overlap. Thus, our analyses provide evidence for an important link between the evolutionary diversification of primates and the richness of their parasite communities, but other mechanisms, particularly those related to reciprocal selection or coextinction of hosts and parasites, require further investigation. PMID- 15540146 TI - Evolution in a changing environment: a case study with great tit fledging mass. AB - Heritable phenotypic traits under significant and consistent directional selection often fail to show the expected evolutionary response. A potential explanation for this contradiction is that because environmental conditions change constantly, environmental change can mask an evolutionary response to selection. We combined an "animal model" analysis with 36 years of data from a long-term study of great tits (Parus major) to explore selection on and evolution of a morphological trait: body mass at fledging. We found significant heritability of this trait, but despite consistent positive directional selection on both the phenotypic and the additive genetic component of body mass, the population mean phenotypic value declined rather than increased over time. However, the mean breeding value for body mass at fledging increased over time, presumably in response to selection. We show that the divergence between the response to selection observed at the levels of genotype and phenotype can be explained by a change in environmental conditions over time, that is, related both to increased spring temperature before breeding and elevated population density. Our results support the suggestion that measuring phenotypes may not always give a reliable impression of evolutionary trajectories and that understanding patterns of phenotypic evolution in nature requires an understanding of how the environment has itself changed. PMID- 15540147 TI - Effects of matrix heterogeneity on animal dispersal: from individual behavior to metapopulation-level parameters. AB - Mounting theoretical and empirical evidence shows that matrix heterogeneity may have contrasting effects on metapopulation dynamics by contributing to patch isolation in nontrivial ways. We analyze the movement properties during interpatch dispersal in a metapopulation of Iberian lynx (Lynx pardinus). On a daily temporal scale, lynx habitat selection defines two types of matrix habitats where individuals may move: open and dispersal habitats (avoided and used as available, respectively). There was a strong and complex impact of matrix heterogeneity on movement properties at several temporal scales (hourly and daily radiolocations and the entire dispersal event). We use the movement properties on the hourly temporal scale to build a simulation model to reconstruct individual dispersal events. The two most important parameters affecting model predictions at both the individual (daily) and metapopulation scales were related to the movement capacity (number of movement steps per day and autocorrelation in dispersal habitat) followed by the parameters representing the habitat selection in the matrix. The model adequately reproduced field estimates of population level parameters (e.g., interpatch connectivity, maximum and final dispersal distances), and its performance was clearly improved when including the effect of matrix heterogeneity on movement properties. To assume there is a homogeneous matrix results in large errors in the estimate of interpatch connectivity, especially for close patches separated by open habitat or corridors of dispersal habitat, showing how important it is to consider matrix heterogeneity when it is present. Movement properties affect the interaction of dispersing individuals with the landscape and can be used as a mechanistic representation of dispersal at the metapopulation level. This is so when the effect of matrix heterogeneity on movement properties is evaluated under biologically meaningful spatial and temporal scales. PMID- 15540148 TI - New insights into chimpanzees, tools, and termites from the Congo Basin. AB - The tool-using behaviors of wild chimpanzees comprise the most impressive assemblages and flexible repertoires of nonhuman material culture. We expand knowledge of the breadth and complexity of tool use in this species by providing the first descriptions of the form and function of two distinct tool sets used by chimpanzees in preying upon termites within the forests of the Goualougo Triangle, Republic of Congo. Further, we report the first application of remote video monitoring technology to record wild chimpanzee tool-using behavior. Based on tool assemblages recovered at termite nests, we hypothesized that chimpanzees were regularly visiting two forms of termite nests and using specific tools to extract termite prey depending on the structure of the nest. Six months of continuous remote video monitoring at six termite nests confirmed that chimpanzees use a tool set to puncture and fish at subterranean termite nests and another tool set to perforate and fish at epigeal (aboveground) nests. Our findings of strict adherence to tool forms at different nest types, tool material selectivity, repeated visits to nests with reusable wood tool assemblages, and differences in material culture between communities have broad implications for our understanding of the ecological and cultural factors that shape hominoid tool use. PMID- 15540149 TI - Spatial structure, dispersal, and management of a recovering raptor population. AB - The recovery of the peregrine falcon (Falco peregrinus anatum) in California has taken place amid strong geographical differences in habitat quality, potentially creating a sink population in the southern coastal habitat and source populations in the northern interior and urban habitats. We analyzed long-term monitoring data to investigate the mechanisms and consequences of spatial structuring for the recovery of this set of nonstable subpopulations. Dispersal rates between habitats were asymmetric, with extremely limited dispersal out of the interior habitat and a strong tendency for birds in the southern coast to disperse to the urban habitats. We used these dispersal estimates and habitat-specific productivity rates to build a set of regional population models that describe population growth within and dispersal between each subpopulation. We tested for the existence of habitat-specific survival and territory acquisition rates by comparing model projections with the number of breeding pairs censused annually in each subpopulation. Our analyses indicate a high rate of survival for interior birds and suggest that both the interior and urban subpopulations were regulated by territory availability over the study period. The inherent spatial structure of this regional peregrine falcon population has had a considerable influence on its recovery and management. PMID- 15540150 TI - The evolution of mimicry under constraints. AB - The resemblance between mimetic organisms and their models varies from near perfect to very crude. One possible explanation, which has received surprisingly little attention, is that evolution can improve mimicry only at some cost to the mimetic organism. In this article, an evolutionary game theory model of mimicry is presented that incorporates such constraints. The model generates novel and testable predictions. First, Batesian mimics that are very common and/or mimic very weakly defended models should evolve either inaccurate mimicry (by stabilizing selection) or mimetic polymorphism. Second, Batesian mimics that are very common and/or mimic very weakly defended models are more likely to evolve mimetic polymorphism if they encounter predators at high rates and/or are bad at evading predator attacks. The model also examines how cognitive constraints acting on signal receivers may help determine evolutionarily stable levels of mimicry. Surprisingly, improved discrimination abilities among signal receivers may sometimes select for less accurate mimicry. PMID- 15540151 TI - Maintenance of trophic structure in fossil mammal communities: site occupancy and taxon resilience. AB - Commonness of organisms typically fluctuates through time, and understanding such fluctuations has long been an important part of ecological research. Studies at paleontological timescales provide a perspective on how changes in population size might affect community structure over millions of years. To overcome the obstacle that population size parameters such as abundance are difficult to detect in the fossil record, we here used fossil locality coverage to approximate changes in site occupancy in the Neogene of Europe over the course of 20 million years. Our aim was to examine whether the trophic structure of mammalian communities is maintained through time despite continuing environmental change. Ecomorphological grouping of fossils indicates that herbivore genera have low taxon resilience in that each genus has predominantly only one locality coverage peak before disappearance. Despite this continuous replacement of the most prevalent herbivore genera, the herbivore trophic group as a whole remains the largest and maintains a roughly constant share of locality coverage throughout the Neogene. The successive herbivore genera, however, show increasing adaptations to harshening environments, indicating shifting properties of niches while the overall trophic structure is conserved. Carnivores, dependent on primary productivity for food indirectly through their prey, show moderate lack of resilience. In contrast, in omnivores changes in locality coverage are close to random fluctuations. PMID- 15540152 TI - Soil water balance and ecosystem response to climate change. AB - Some essential features of the terrestrial hydrologic cycle and ecosystem response are singled out by confronting empirical observations of the soil water balance of different ecosystems with the results of a stochastic model of soil moisture dynamics. The simplified framework analytically describes how hydroclimatic variability (especially the frequency and amount of rainfall events) concurs with soil and plant characteristics in producing the soil moisture dynamics that in turn impact vegetation conditions. The results of the model extend and help interpret the classical curve of Budyko, which relates evapotranspiration losses to a dryness index, describing the partitioning of precipitation into evapotranspiration, runoff, and deep infiltration. They also provide a general classification of soil water balance of the world ecosystems based on two governing dimensionless groups summarizing the climate, soil, and vegetation conditions. The subsequent analysis of the links among soil moisture dynamics, plant water stress, and carbon assimilation offers an interpretation of recent manipulative field experiments on ecosystem response to shifts in the rainfall regime, showing that plant carbon assimilation crucially depends not only on the total rainfall during the growing season but also on the intermittency and magnitude of the rainfall events. PMID- 15540153 TI - How the spatial scales of dispersal, competition, and environmental heterogeneity interact to affect coexistence. AB - Spatial coexistence depends on a variety of biological and physical processes, and the relative scales of these processes may promote or suppress coexistence. We model plant competition in a spatially varying environment to show how shifting scales of dispersal, competition, and environmental heterogeneity affect coexistence. Spatial coexistence mechanisms are partitioned into three types: the storage effect, nonlinear competitive variance, and growth-density covariance. We first describe how the strength of each of these mechanisms depends on covariances between population densities and between population densities and the environment, and we then explain how changes in the scales of dispersal, competition, and environmental heterogeneity should affect these covariances. Our quantitative approach allows us to show how changes in the scales of biological and physical processes can shift the relative importance of different classes of spatial coexistence mechanisms and gives us a more complete understanding of how environmental heterogeneity can enable coexistence. For example, we show how environmental heterogeneity can promote coexistence even when competing species have identical responses to the environment. PMID- 15540154 TI - An experimental test of the dose-dependent effect of carotenoids and immune activation on sexual signals and antioxidant activity. AB - Carotenoid-based sexual traits are thought to be reliable indicators of male quality because they might be scarce and therefore might indicate the ability of males to gather high-quality food and because they are involved in important physiological functions (as immune enhancers and antioxidants). We performed an experiment where male and female zebra finches (Taeniopygia guttata) were provided with increasing carotenoid doses in the drinking water during 4 weeks (bill color of this species is a carotenoid-based sexual signal). Simultaneously, birds were split into two groups: one receiving weekly injections of Escherichia coli lipopolysaccharide in order to activate the immune system, the other being injected with the same volume of phosphate buffered saline. We assessed how carotenoid availability and immune activation affected the amount of circulating plasma carotenoids, the beak color, and the antioxidant defenses (assessed as the resistance of red blood cells to a controlled free radical attack). Carotenoid availability affected the amount of circulating carotenoids and beak color; both variables reached a plateau at the highest carotenoid doses. Immune activation diverted carotenoids from plasma, and this in turn affected the expression of the sexual trait. Finally, we found a positive correlation between the change in circulating carotenoids and antioxidant defenses. These results support the idea that carotenoids have important physiological properties that ensure the honesty of carotenoid-based sexual traits. PMID- 15540155 TI - Genetic and environmental influences on natal dispersal distance in a resident bird species. AB - We analyzed more than 1,600 dispersal events from two populations of a North American cooperatively breeding woodpecker species to determine what factors influence natal dispersal distance and whether distance traveled affects reproduction later in life. We found significant heritability of natal dispersal distance, in both males and females, indicating substantial additive genetic variance for this behavioral trait. Natal dispersal distance additionally was affected by social and ecological factors: individuals dispersing in their first year of life moved longer distances than those staying on their natal site as helpers for a prolonged time prior to dispersal, and increasing territory isolation led to longer dispersal distances. Successful dispersers incurred fitness costs, with lifetime fledgling production (in both sexes) and lifetime production of recruits to the breeding population (in females only) decreasing with increasing natal dispersal distance. We conclude that natal dispersal distance has a genetic basis but is modulated by environmental and social factors and that natal dispersal distance in this species is (currently) under selection. PMID- 15540156 TI - Trade-offs in community properties through time in a desert rodent community. AB - Resource limitation represents an important constraint on ecological communities, which restricts the total abundance, biomass, and community energy flux a given community can support. However, the exact relationship among these three measures of biological activity remains unclear. Here we use a simple framework that links abundance and biomass with an energetic constraint. Under constant energetic availability, it is expected that changes in abundance and biomass can result from shifts in the distribution of individual masses. We test these predictions using long-term data from a desert rodent community. Total energy use for the community has not changed directionally for 25 years, but species composition has. As a result, the average body size has decreased by almost 50%, and average abundance has doubled. These results lend support to the idea of resource limitation on desert rodent communities and demonstrate that systems are able to maintain community energy flux in the face of environmental change, through changes in composition and structure. PMID- 15540157 TI - Normalized mutual entropy in biology: quantifying division of labor. AB - Division of labor is one of the primary adaptations of sociality and the focus of much theoretical work on self-organization. This work has been hampered by the lack of a quantitative measure of division of labor that can be applied across systems. We divide Shannon's mutual entropy by marginal entropy to quantify division of labor, rendering it robust over changes in number of individuals or tasks. Reinterpreting individuals and tasks makes this methodology applicable to a wide range of other contexts, such as breeding systems and predator-prey interactions. PMID- 15540158 TI - Mathematical assumptions versus biological reality: myths in affected sib pair linkage analysis. AB - Affected sib pair (ASP) analysis has become common ever since it was shown that, under very specific assumptions, ASPs afford a powerful design for linkage analysis. In 2003, Vieland and Huang, on the basis of a "fundamental heterogeneity equation," proved that heterogeneity and epistasis are confounded in ASP linkage analysis. A much more serious limitation of ASP linkage analysis is the implicit assumption that randomly sampled sib pairs share half their alleles identical by descent at any locus, whereas a critical assumption underlying Vieland and Huang's proof is that of joint Hardy-Weinberg equilibrium proportions at two trait loci. These are considered as examples of mathematical assumptions that may not always reflect biological reality. More-robust sib-pair designs and appropriate methods for their analysis have long been available. PMID- 15540159 TI - Prospects for admixture mapping of complex traits. AB - Admixture mapping extends to human populations the principles that underlie linkage analysis of an experimental cross. For detecting genes that contribute to ethnic variation in disease risk, admixture mapping has greater statistical power than family-linkage studies. In comparison with association studies, admixture mapping requires far fewer markers to search the genome and is less affected by allelic heterogeneity. Statistical-analysis programs for admixture mapping are now available, and a genomewide panel of markers for admixture mapping in populations formed by West African-European admixture has been assembled. Some of the remaining technical challenges include the ability to ensure that the statistical methods are robust and to develop marker panels for other admixed populations. Where admixed populations and panels of markers informative for ancestry are available, admixture mapping can be applied to localize genes that contribute to ethnic variation in any measurable trait. PMID- 15540160 TI - Mapping a Mendelian form of intracranial aneurysm to 1p34.3-p36.13. AB - The identification of pathways that underlie common disease has been greatly impacted by the study of rare families that segregate single genes with large effect. Intracranial aneurysm is a common neurological problem; the rupture of these aneurysms constitutes a frequently catastrophic neurologic event. The pathogenesis of these aneurysms is largely unknown, although genetic and environmental factors are believed to play a role. Previous genomewide studies in affected relative pairs have suggested linkage to several loci, but underlying genes have not been identified. We have identified a large kindred that segregates nonsyndromic intracranial aneurysm as a dominant trait with high penetrance. Genomewide analysis of linkage was performed using a two-stage approach: an analysis of ~10,000 single-nucleotide polymorphisms in the 6 living affected subjects, followed by the genotyping of simple tandem repeats across resulting candidate intervals in all 23 kindred members. Analysis revealed significant linkage to a single locus, with a LOD score of 4.2 at 1p34.3-p36.13 under a dominant model with high penetrance. These findings identify a Mendelian form of intracranial aneurysm and map the location of the underlying disease locus. PMID- 15540161 TI - Upregulation of Wilms' tumor gene 1 (WT1) in desmoid tumors. AB - Desmoid tumors (aggressive fibromatosis) are locally invasive soft tissue tumors in which beta-catenin/TCF3 mediated Wnt signaling is activated. More than 80% of desmoid tumors contain activating mutations in beta-catenin. It has been shown that the Wnt signaling pathway interacts with Wilms' tumor gene 1 (WT1) in normal kidney development and plays a role in the genesis of some Wilms' tumors. About 15% of Wilms' tumors contain WT1 mutations and of these, about 50% contain beta catenin mutations. This overlap in mutation pattern of WT1 and beta-catenin in Wilms' tumor suggests that these 2 genes may collaborate in the genesis of a subset of Wilms' tumors. To investigate whether this hypothesis could be extended to other Wnt-dependent tumor types, we searched for WT1 mutations and studied WT1 expression in beta-catenin mutant desmoid tumors. We investigated the expression of WT1 mRNA and protein in desmoid tumors. Medium to high abundant levels of WT1 mRNA were detected by TaqMan quantitative PCR in all tested desmoid cells, whereas adjacent normal fibroblasts showed less expression of WT1. Western blot analysis and immunohistochemistry confirmed this overexpression at the protein level. A mutational screen of the WT1 zinc-finger region by sequence analysis did not identify any mutations. Finally, we investigated a possible role of beta catenin on WT1 regulation and vice versa. Overexpression of different beta catenin mutants in the HEK293T cell line did not modulate WT1 promoter activity and WT1 did not affect beta-catenin /TCF transcriptional activity in this cell line. These results show that the wild-type WT1 gene is strongly overexpressed in beta-catenin mutant desmoid tumors and may play a role in tumorigenesis of desmoid tumors, similar to what has been suggested in some epithelial malignancies. PMID- 15540163 TI - A combination of protein profiling and isotopomer analysis using matrix-assisted laser desorption/ionization-time of flight mass spectrometry reveals an active metabolism of the extracellular matrix of 3T3-L1 adipocytes. AB - Differential gel electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is a commonly used protein profiling method. However, observed changes can be explained in multiple ways, one of which is by the protein turnover rate. In order to easily and rapidly obtain information on both the identity and turnover of individual proteins, we applied a combination of protein labeling with L-(ring 2,3,4,5,6 2H5) phenylalanine and MALDI-TOF MS. While the spectrum reveals the identity of a protein, mass isotopomer analysis provides information about the rate of protein labeling as a measure of synthesis or turnover. Using this approach on mature 3T3-L1 adipocytes, we were able to discriminate between rapidly and slowly metabolised proteins. In our isolate, proteins of the cytoskeleton appeared to be slowly metabolised, whereas components of the extracellular matrix, in particular collagen type I alpha 1 (COL1A1) and collagen type I alpha 2 (COL1A2) showed rapid accumulation of newly synthesized proteins. Both proteins appeared to be metabolised in the same ratio as they are present in collagen fibers, i.e. 2:1 (COL1A1: COL1A2). In addition, functionally related proteins were also readily labeled. Taken together, we have shown that a combination of stable isotope labeling and protein profiling by gel electrophoresis and MALDI-TOF analysis can simultaneously provide information on the identity and relative metabolic rate of proteins in eukaryotic cells in a simple, nonhazardous and rapid-throughput way. PMID- 15540164 TI - Fine mapping of chromosome 10q deletions in mycosis fungoides and sezary syndrome: identification of two discrete regions of deletion at 10q23.33-24.1 and 10q24.33-25.1. AB - Previous cytogenetic studies in mycosis fungoides (MF) and Sezary syndrome (SS) have identified a large and poorly defined area of chromosomal deletion on chromosome 10q. We report an extensive fine-mapping allelotyping study using 19 microsatellite markers in the region 10q22.3-10q26.13. Allelic loss was identified by loss of heterozygosity analysis in 26 of 60 (43%) cases: 15 of 45 (33%) with MF and 11 of 15 (73%) with SS. MF and SS samples showed similar patterns of allelic loss with the identification of two discrete regions of deletion which were mutually exclusive in all but two cases. Within the first region of deletion at 10q23.33-10q24.1, around microsatellite marker D10S185 (2.77 Mb), 23 genes were identified, including three (KIF11, HHEX, and HELLS) with functions that, if dysregulated, could be critical in MF and SS. The second region of deletion, 10q24.33-10q25.1, around microsatellite marker D10S530 (3.92 Mb), encodes 11 genes, the majority of which have poorly identified functions. This extensive allelotyping study provides the basis for future highly selective candidate gene analyses. PMID- 15540165 TI - Gene amplifications in osteosarcoma-CGH microarray analysis. AB - Little is known about the genomic alterations underlying osteosarcoma. We performed a genomewide high-resolution gene copy number analysis of 22 osteosarcoma samples using comparative genomic hybridization on a cDNA microarray that contained cDNA clones of about 13,000 genes. Nineteen of the 22 cases had amplifications that on average spanned more than 1 Mb and contained more than 10 genes. Numerous regions of gain and loss were identified, and their boundaries were defined at high resolution. Novel amplicons were found at 14q11, 17q25, and 22q11-q13. Earlier-known large amplified regions were detected at 12q11-q15, 8q24, 6p12-p13, and 17p11-p13 in 8, 6, 5, and 4 of the 22 samples, respectively. Amplification of 12q was observed more frequently (36% of the cases) than previously reported. Previously known small amplicons at 1p34-p36, 1q21, 19q13, and 21q22 were seen in at least three cases. Our results implicate TOM1L2 and CYP27B1 as having roles as novel targets for the 17p and 12q amplicons, respectively. Details (www.helsinki.fi/cmg) of the amplified genes in each amplicon provide valuable raw data for further in silico studies. PMID- 15540166 TI - Identification of protein-protein interactions using in vivo cross-linking and mass spectrometry. AB - The purification of protein complexes can be accomplished by different types of affinity chromatography. In a typical immunoaffinity experiment, protein complexes are captured from a cell lysate by an immobilized antibody that recognizes an epitope on one of the known components of the complex. After extensive washing to remove unspecifically bound proteins, the complexes are eluted and analyzed by mass spectrometry (MS). Transient complexes, which are characterized by high dissociation constants, are typically lost by this approach. In the present study, we describe a novel method for identifying transient protein-protein interactions using in vivo cross-linking and MS-based protein identification. Live cells are treated with formaldehyde, which rapidly permeates the cell membrane and generates protein-protein cross-links. Proteins cross-linked to a Myc-tagged protein of interest are copurified by immunoaffinity chromatography and subjected to a procedure which dissociates the cross-linked complexes. After separation by SDS-PAGE, proteins are identified by tandem mass spectrometry. Application of this method enabled the identification of numerous proteins that copurified with a constitutively active form of M-Ras (M Ras(Q71L)). Among these, we identified the RasGAP-related protein IQGAP1 to be a novel interaction partner of M-Ras(Q71L). This method is applicable to many proteins and will aid in the study of protein-protein interactions. PMID- 15540167 TI - Acidic proteome of growing and resting Lactococcus lactis metabolizing maltose. AB - The acidic proteome of Lactococcus lactis grown anaerobically was compared for three different growth conditions: cells growing on maltose, resting cells metabolizing maltose, and cells growing on glucose. In maltose metabolizing cells several proteins were up-regulated compared with glucose metabolizing cells, however only some of the up-regulated proteins had apparent relation to maltose metabolism. Cells growing on maltose produced formate, acetate and ethanol in addition to lactate, whereas resting cells metabolizing maltose and cells growing on glucose produced only lactate. Increased levels of alcohol-acetaldehyde dehydrogenase (ADH) and phosphate acetyltransferase (PTA) in maltose-growing cells compared with glucose-growing cells coincided with formation of mixed acids in maltose-growing cells. The resting cells did not grow due to lack of an amino acid source and fermented maltose with lactate as the sole product, although ADH and PTA were present at high levels. The maltose consumption rate was approximately three times lower in resting cells than in exponentially growing cells. However, the enzyme levels in resting and growing cells metabolizing maltose were similar, which indicates that the difference in product formation in this case is due to regulation at the enzyme level. The levels of 30S ribosomal proteins S1 and S2 increased with increasing growth rate for resting cells metabolizing maltose, maltose-growing cells and glucose-growing cells. A modified form of HPr was synthesized under amino acid starvation. This is suggested to be due to alanine misincorporation for valine, which L. lactis is auxotrophic for. L. lactis conserves the protein profile to a high extent, even after prolonged amino acid starvation, so that the protein expression profile of the bacterium remains almost invariant. PMID- 15540170 TI - Improved mass spectrometric identification of gel-separated hydrophobic membrane proteins after sodium dodecyl sulfate removal by ion-pair extraction. AB - Separation and identification of hydrophobic membrane proteins is a major challenge in proteomics. Identification of such sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)-separated proteins by peptide mass fingerprinting (PMF) via matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) is frequently hampered by the insufficient amount of peptides being generated and their low signal intensity. Using the seven helical transmembrane-spanning proton pump bacteriorhodopsin as model protein, we demonstrate here that SDS removal from hydrophobic proteins by ion-pair extraction prior to in-gel tryptic proteolysis leads to a tenfold higher sensitivity in mass spectrometric identification via PMF, with respect to initial protein load on SDS-PAGE. Furthermore, parallel sequencing of the generated peptides by electrospray ionization-mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) was possible without further sample cleanup. We also show identification of other membrane proteins by this protocol, as proof of general applicability. PMID- 15540171 TI - Fine-needle aspiration of primary and recurrent benign fibrous histiocytoma: classic, aneurysmal, and myxoid variants. AB - There is a limited number of correlative cytopathological studies of fibrous histiocytoma (FHC). To better define cytopathological criteria of diagnosis, we have reviewed fine-needle aspirates (FNA) from 36 FHCs (32 classical, 1 myxoid, and 3 aneurysmal variants on corresponding histological sections). Original cytological diagnoses were benign in 33 (91.7%) cases (22 accurate) and false positive in 3 (8.3%) cases. All smears were surprisingly homogenous and composed of histiocytic cells with finely vacuolated cytoplasm in 27 (75%) cases, small regular spindle cells in 25 (69%) cases, and giant cells in 17 (47%) cases. Histiocytic cells were attached to vascular structures in 9 (25%) cases. Slight cytonuclear atypia was seen in five (14%) cases. Three (8.3%) cases showed numerous siderophages. In two (5.6%) cases, there were abundant inflammatory backgrounds and in one (3%) case there was a scant myxoid background. Storiform patterns, round cells, prominent atypia, necroses, or mitotic figures were not seen. FHC should be differentiated from other benign, low- and intermediate-grade spindle-cell neoplasms such as low-grade fibrosarcoma, dermatofibrosarcoma protuberans, nodular fasciitis, spindle-cell malignant melanoma, and monophasic synovial sarcoma. Some cases may be misinterpreted as malignant, especially in cases of recurrence or in patients with a cancer history. PMID- 15540172 TI - A rare case of primary malignant melanoma of the scrotum diagnosed by fine-needle aspiration. AB - We report a case of a 55-yr-old man with malignant melanoma of scrotum. He was referred to our Hospital with a complaint of gradual focal enlargement of the scrotum in a period of 3 yr. On physical examination, a pigmented, poorly marginated mass, with central necrosis was observed. A fine-needle aspiration (FNA) of the lesion was performed. Cytological examination revealed highly cellular smears, containing malignant cells, dispersed or arranged in loose aggregates. Cellular morphology and characteristics were identical to those of malignant melanoma arising elsewhere in the skin. The immunocytochemical study revealed positivity of neoplastic cells for anti-melanoma monoclonal antibody (HMB-45) antigen. Histological confirmation finally was provided after wide excision of the lesion. We emphasize the difficulties in differential diagnosis considerations and diagnostic pitfalls of scrotal lesions. PMID- 15540173 TI - Columnar cell lesions: fine-needle aspiration biopsy features. AB - Columnar cell lesions (CCLs) have been described histologically. Frequently, they are noted in biopsies performed for calcifications and are associated with an increased risk of malignancy in the presence of atypia. We sought to characterize the cytological features of CCLs in fine-needle aspirations (FNAs). Twenty FNAs with subsequent histology diagnoses of CCL without carcinoma were reviewed retrospectively. Eighteen of 20 cases were classified as "atypical" on cytology; they had cohesive three-dimensional clusters of enlarged polygonal epithelial cells intermixed with myoepithelial cells in the centers and palisading columnar cells peripherally. Five of these had cytological or architectural atypia on subsequent biopsies, but no significant differences were noted among the 18 aspirates. Calcifications (2/18), snouts (9/18), and bipolar nuclei (11/18) were also identified. The remaining 2/20 FNAs were interpreted as negative because of scant cellularity. In conclusion, CCLs have characteristic cytological traits, and because they may be associated with carcinoma, their recognition is important. PMID- 15540174 TI - Assessing the feasibility of single lifetime PAP smear evaluation between 41-50 years of age as strategy for cervical cancer control in developing countries from our 32 years of experience of hospital-based routine cytological screening. AB - In view of the huge expenditure involved in mass cytological screening as well as lack of cytology manpower in the developing countries, single lifetime screening at 45 yr of age has been suggested as feasible strategy for control of cervical cancer. The present study is aimed at testing this hypothesis in a broader prospective, accommodating women between 41 and 50 yr of age from the data derived from the ongoing long-term hospital-based routine cervicovaginal cytology at Queen Mary's Hospital, Lucknow, India. The cervical smears of 31,032 women have been evaluated cytologically during a span of 32 yr (April 1971-March 2003) for early detection of carcinoma cervix and sexually transmitted diseases (STDs). The incidence of squamous intraepithelial lesions (SIL) was found to be 6.1% while frank malignancy was seen in 0.5% of cases. The following observations were made from the accumulated cytological data: (a) approximately 30% of the total cancer cases was in the age group of 41-50 yr; (b) the maximum number of SIL cases was detected in women between 41 and 50 yr of age (35% of the total SIL cases), and adequate management of these SIL cases would prevent cancer cases from occurring in later years, thus minimizing the maximum incidence of 1.3% observed in women beyond 50 yr of age; (c) the incidence of human papillomavirus (HPV) infection also was found quite high in women between 41 and 50 yr old, the adequate management of which would prevent onset/progression of any premalignant changes in the cervix. Our experience of 32 yr of cytological screening substantiates the hypothesis of single lifetime screening between 41 and 50 yr of age as an effective strategy for control of carcinoma cervix in developing countries like ours. PMID- 15540175 TI - Opitz "C" trigonocephaly-like syndrome in a patient with terminal deletion of 2p and partial duplication of 17q. AB - A boy with trigonocephaly, cleft palate, multiple minor anomalies, flexion deformities of elbows, cryptorchidism, and severe muscular hypotonia had an unbalanced karyotype with duplication of the distal 17q and deletion of the tip of 2p. This was derived from a reciprocal translocation in the father, 46,XY,t(2;17)(p25;q24). The propositus had some findings observed in patients with distal dup(17q), while trigonocephaly not found in these patients may be associated with the terminal deletion of 2p including the locus of SOX11 gene. It is proposed that the major clinical findings of this patient are consistent with the phenotype characteristic of the Opitz "C" trigonocephaly syndrome. PMID- 15540176 TI - Precision in phenotyping and genotyping. PMID- 15540177 TI - Immunostaining of galectin-3 and CD44v6 using fine-needle aspiration for distinguishing follicular carcinoma from adenoma. AB - To evaluate the clinical applicability of galectin-3 and CD44 variant 6 (CD44v6) immunostaining in fine-needle aspiration cytology (FNAC) of thyroid follicular tumors, 79 cytological specimens (35 follicular carcinomas and 44 follicular adenomas) were studied. The positive rates of galectin-3 and CD44v6 were 89 and 74% in follicular carcinoma, respectively, and 25 and 30% in follicular adenoma, respectively. There were no significant correlations between the expression of galectin-3 or CD44v6 in follicular carcinoma and characteristics such as capsular invasion, vascular invasion, metastasis, or tumor size. Positive staining of either galectin-3 or CD44v6 resulted in a diagnostic sensitivity of 97% and a specificity of 52% for follicular carcinoma among follicular tumors. Immunostaining of galectin-3 or CD44v6 using cytological specimens can provide independent information on conventional morphological findings of cytology to distinguish follicular carcinoma from adenoma. PMID- 15540178 TI - Chronic endometritis: an added diagnostic value for Pap smear. PMID- 15540179 TI - Chondrosarcoma of calcanaeum in a 12-year-old male patient: a case report. AB - Chondrosarcoma is distinctly uncommon and tends to be located often in the extremities in young patients more than in its adult counterpart. Cartilaginous tumors involving the small bones of the hands and feet are benign tumors such as enchondromas, chondromyxoid fibroma (CMF), and chondroblastoma. Chondrosarcomas involving calcanaeum in young adults are largely covered in the literature as single case reports. A young 12-yr-male patient presented with complaints of pain and mild swelling in the ankle. Radiological examination revealed a dense irregular lesion in the calcanaeum. Fine-needle aspiration was performed and a possibility of chondrosarcoma was suspected, which later was confirmed on histopathological examination. The cytopathologist should be aware of the occurrence of malignant chondroid tumors in the younger age group at this rare site. Correlation to radiological findings is essential in these cases. PMID- 15540180 TI - Recurrent respiratory papillomatosis: cytopathological findings in an unusual case. AB - Recurrent respiratory papillomatosis (RRP) primarily is a disease of children and adolescents. Most patients experience spontaneous regression at a later age, while others continue to be affected throughout their lives. Here, we present the case of a 26-yr-old man who presented with persistent cough, worsening dyspnea, and development of pneumothorax. Eight years prior, he had presented with similar complaints and an evaluation at that time had showed RRP. Follow-up since that time showed recurrent disease managed by multiple surgical procedures. At the most current admission, an ultrasound-guided fine-needle aspiration (FNA) of the lung showed numerous papillary tissue fragments and single atypical squamous cells consistent with recurrent RPR. To our knowledge, morphological findings of RRP have been reported rarely. PMID- 15540181 TI - Fine-needle aspiration diagnosis of plasmacytoma presenting as breast masses in a patient on estrogen therapy for prostate cancer. AB - We describe a 79-yr-old man with a history of androgen-independent metastatic prostate cancer treated with exogenous estrogens presenting with bilateral breast masses associated with bilateral axillary lymphadenopathy. Although the findings on physical examination with the concomitant history of estrogen therapy for metastatic prostate cancer raised the clinical suspicion of breast cancer, fine needle aspiration (FNA) cytology identified the lesions as multiple myeloma. PMID- 15540182 TI - Erdheim-Chester disease of the brain: cytological features and differential diagnosis of a challenging case. AB - Erdheim-Chester disease (ECD) is an uncommon, systemic xanthogranulomatous disorder, with distinct clinicopathological features, that is rarely expected preoperatively. We describe a case that presented in the brain of a 26-yr-old male patient and clinically mimicked the appearance of a neoplasm. The final diagnosis was a surprise. In retrospect, the diagnosis was suggested by the intraoperative "squash" preparations, which demonstrated a mixed cellular proliferation of lymphohistiocytic elements and large, multinucleated cells with vesicular nuclei, prominent nucleoli, and abundant cytoplasm. To the best of our knowledge, this is the first report detailing the cytopathological features of ECD. PMID- 15540183 TI - Comparison of three different staining techniques for intraoperative assessment of nodal metastasis in breast cancer. AB - Imprint cytology has increasingly been used for intraoperative assessment of nodal status in breast cancer. We carried out this study to compare the efficacy of Jenner Giemsa (JG), hematoxylin-eosin (H&E), and Papanicolaou (Pap) stains for intraoperative lymph node imprint cytology (IIC) in breast cancer. One hundred and seven cases of stage I-III breast cancer were studied. Overall, IIC was accurate in 95.3% cases and had a sensitivity and specificity of 98.5% and 90.0%, respectively. The accuracy of JG (95.3%) was better than that of H&E (90.6%) and Pap (94.0%), although the differences were not statistically significant. Problems encountered included cell loss and drying artifacts with H&E and Pap and the inability to distinguish between tumor cells and histiocytes confidently in tight cellular clusters that were occasionally seen. Opinion was possible in all JG cases, but not in five and four cases by H&E and Pap, respectively. Although the choice of the stain would vary depending on the experience of the pathologist, our work suggests that JG, because of fewer technical problems and superior accuracy, may be preferable over H&E and Pap. PMID- 15540184 TI - Interpretive yields of screening Pap tests and diagnostic Pap tests. AB - Recently, our laboratory has introduced the Medicare classification "diagnostic Papanicolaou (Pap) test" (DPT) for both conventional and liquid-based Pap tests. Because little is known regarding the diagnostic yield of DPT, we review our experience with these tests in a community with a low incidence of squamous intraepithelial lesions and contrast this with our experience with screening Pap tests (SPTs). A search of our laboratory's computerized data system identified all Pap tests classified as SPT and DPT from December 3, 2002 to December 2, 2003. The interpretations were tabulated and statistical comparisons were made. Between December 3, 2002 and December 2, 2003 63,626 Pap tests were interpreted (57,922 SPTs and 5704 DPTs). DPTs were far more likely to yield abnormal results (atypical squamous cells or worse, P<0.001) and were especially more likely to yield malignant results. These differences have numerous possible implications regarding screening, rescreening, and the design of studies that investigate the screening value of the Pap test. PMID- 15540186 TI - The emerging biological and clinical significance of the columnar cell lesions of the breast. PMID- 15540187 TI - Psammoma body and its precursors in papillary thyroid carcinoma: a study by fine needle aspiration cytology. AB - Psammoma bodies (PBs) form an important diagnostic criterion of papillary thyroid carcinoma (PTC), but their mechanism of formation is not clear. Following our observation in a case of PTC that laminated hyaline globules may be the precursor form of PBs, the present study was undertaken to find out their relation to PBs in PTC cases. Fifty-four cases of PTC and 14 follicular neoplasms, diagnosed by fine-needle aspiration (FNA) cytology, were studied by one of the investigators (DKD) to find out the PBs, irregular calcifications, hyaline globules, and other forms made of similar material. PBs along with irregular calcification were present in five PTC cases, PB alone was present in 1 case, and irregular calcification alone was present in 4 cases. Large hyaline globules (LHGs), small hyaline globules (SHGs), branching hyaline cylinders (BHCs), and irregular hyaline deposits (IHDs) were identified in 10, 14, 6, and 9 cases, respectively. One or more of these four forms were present altogether in 18 (33.3%) of PTC cases and none of the follicular neoplasms (P=0.0142). These forms were present in 80.0% of cases with PB/irregular calcifications as opposed to 22.7% in cases without them (P=0.0012). Our observations suggest that LHGs, SHGs, and BHCs are precursors of PBs and IHDs serve as a nidus for irregular calcification. PMID- 15540188 TI - The fine-needle aspiration biopsy cytology of pancreatoblastoma. AB - We describe the clinical, cytological, histological, and immunohistochemical features of primary and metastatic pancreatoblastoma (PBL), a rare primary pancreatic malignancy of both adults and children. An 18-yr-old male patient presented with abdominal pain, weight loss, and diarrhea and had a 9-cm pancreatic mass that was revealed by CT scan. The fine-needle aspiration biopsy (FNAB) smears from the pancreas specimen and the lung metastasis were cellular and were composed of a combination of solid sheets, three-dimensional, loosely cohesive epithelial groups, and primitive spindled mesenchymal tissue with focal cartilage formation. Acinar formations were best appreciated on cell block preparations, and squamoid corpuscles were seen only on cell block. Zymogen granules were demonstrated in a subset of the epithelial cells using periodic acid Schiff stains with and without diastase and dual acinar-endocrine differentiation was evident using a focused panel of immunohistochemical stains. Acinar-cell carcinoma (ACC) is the most difficult neoplasm to distinguish from PBL, both clinically and cytologically, especially in young children. The key to distinguishing them is to note the presence of squamous corpuscles and/or heterologous elements such as cartilage. Given the increasing utility of FNAB in the investigation of pancreatic masses, it is important for the pathologist to be familiar with the morphological features of this tumor, especially because preoperative diagnosis provides an opportunity for preoperative therapy before resection. PMID- 15540189 TI - Secretory carcinoma in a 9-year-old girl. PMID- 15540190 TI - Fine-needle aspiration cytodiagnosis of nipple adenoma (papillomatosis) in a man and woman. PMID- 15540192 TI - Modulation by flavonoids of DNA damage induced by estrogen-like compounds. AB - Reactive oxygen species (ROS) are produced by a wide variety of exogenous chemicals and metabolic processes and cause a broad spectrum of damage to biological systems. As a consequence, ROS react with DNA, among many other biological targets, disrupting its structure and functionality. Estrogen-like compounds mediate DNA damage by ROS generation, implying that their effects can be modulated by antioxidants such as catalase, superoxide dismutase, and vitamin C. We examined DNA damage in human lymphocytes and sperm after treatment with four estrogen-like compounds (beta-estradiol, diethylstilbestrol, daidzein, and genistein) and its modulation by flavonoids (quercetin and kaempferol) using the Comet assay. The results indicated that quercetin and kaempferol reduced the DNA damage produced in sperm and lymphocytes by the four estrogenic compounds. The flavonoids also reduced the DNA damage induced by hydrogen peroxide, which was used as a positive control. Our results demonstrate that the antioxidant properties of flavonoids can protect the integrity of human sperm and lymphocyte DNA from ROS induced by estrogenic compounds. PMID- 15540193 TI - Induction of micronucleated cells in the shed skin of salamanders (Ambystoma sp.) treated with colchicine or cyclophosphamide. AB - The micronucleus (MN) assay can be used to detect the genotoxic effects of chemical agents in virtually any cell that divides frequently. Salamanders (Ambystoma sp.) are amphibians that can be easily maintained and bred in the laboratory and spontaneously shed their skin every 2.5-4 days. In this present study, we have evaluated the usefulness of this shed skin for the MN assay. We exposed salamanders to different concentrations of both the aneugen colchicine (COL) and the clastogen cyclophosphamide (CP) and we determined the frequency of micronucleated cells (MNCs) in their sheds. Fragments of shed skin were placed on clean slides, fixed, stained, observed with a light microscope, and the number of MNCs was counted. The MNC frequency was increased significantly by all doses of COL and CP tested, administered either as single or repeated exposures. The presence of MNCs in the shed skin and the speed of sloughing lead us to propose that the sheds of Ambystoma sp., or other amphibians that slough their skin, are suitable alternative models for detecting genotoxic exposures relevant to aquatic environments. PMID- 15540194 TI - Anaerobic degradation of solid material: importance of initiation centers for methanogenesis, mixing intensity, and 2D distributed model. AB - Batch anaerobic codigestion of municipal household solid waste (MHSW) and digested manure in mesophilic conditions was carried out. The different waste-to biomass ratios and intensity of mixing were studied theoretically and experimentally. The experiments showed that when organic loading was high, intensive mixing resulted in acidification and failure of the process, while low mixing intensity was crucial for successful digestion. However, when loading was low, mixing intensity had no significant effect on the process. We hypothesized that mixing was preventing establishment of methanogenic zones in the reactor space. The methanogenic zones are important to withstand inhibition due to development of acids formed during acidogenesis. The 2D distributed models of symmetrical cylinder reactor are presented based on the hypothesis of the necessity of a minimum size of methanogenic zones that can propagate and establish a good methanogenic environment. The model showed that at high organic loading rate spatial separation of the initial methanogenic centers from active acidogenic areas is the key factor for efficient conversion of solids to methane. The initial level of methanogenic biomass in the initiation centers is a critical factor for the survival of these centers. At low mixing, most of the initiation methanogenic centers survive and expand over the reactor volume. However, at vigorous mixing the initial methanogenic centers are reduced in size, averaged over the reactor volume, and finally dissipate. Using fluorescence in situ hybridization, large irregular cocci of microorganisms were observed in the case with minimal mixing, while in the case with high stirring mainly dead cells were found. PMID- 15540195 TI - Monitoring of active but non-culturable bacterial cells by flow cytometry. AB - Flow cytometric signatures (i.e., light scatter, red and green fluorescence) were obtained for the active but non-culturable (ABNC) cells of E. coli and a coliform isolate H03N1, in seawater microcosms using BacLight, a live-dead assay kit from Molecular Probes (Eugene/Portland, OR). Previous studies have reported that there are two major adaptations, which cells undergo during the formation of ABNC states: cell wall toughening and DNA condensation. Therefore, we hypothesized that the matured ABNC forms should be more resistant to extreme temperature treatments (i.e., by freezing in liquid nitrogen and thawing at room temperature) than the normal and transition populations. It was shown that the membrane compromised cells (comprising of normal wild-type and dead cells which are less resistant to rapid freeze thaw) could be differentiated from the matured ABNC using BacLight staining and fluorescence detection by flow cytometry. The population of ABNC cells, which could not be cultured using m-FC media (for the enumeration of fecal coliforms), was resuscitated in phosphate buffer saline followed by growth in Luria broth. Flow cytometry was thus able to detect and differentiate the ABNC cells against a mixed population comprising of culturable cells, transition populations, and dead cells. The results also showed that the formation of ABNC is as early as 2 days in seawater microcosms. By directly comparing the coliform levels enumerated by the BacLight based flow cytometry assays and m-FC technique, it was shown that the presence of coliforms can be undetected by the membrane filtration method. PMID- 15540196 TI - Inferring gene regulatory relationships by combining target-target pattern recognition and regulator-specific motif examination. AB - Although microarray data have been successfully used for gene clustering and classification, the use of time series microarray data for constructing gene regulatory networks remains a particularly difficult task. The challenge lies in reliably inferring regulatory relationships from datasets that normally possess a large number of genes and a limited number of time points. In addition to the numerical challenge, the enormous complexity and dynamic properties of gene expression regulation also impede the progress of inferring gene regulatory relationships. Based on the accepted model of the relationship between regulator and target genes, we developed a new approach for inferring gene regulatory relationships by combining target-target pattern recognition and examination of regulator-specific binding sites in the promoter regions of putative target genes. Pattern recognition was accomplished in two steps: A first algorithm was used to search for the genes that share expression profile similarities with known target genes (KTGs) of each investigated regulator. The selected genes were further filtered by examining for the presence of regulator-specific binding sites in their promoter regions. As we implemented our approach to 18 yeast regulator genes and their known target genes, we discovered 267 new regulatory relationships, among which 15% are rediscovered, experimentally validated ones. Of the discovered target genes, 36.1% have the same or similar functions to a KTG of the regulator. An even larger number of inferred genes fall in the biological context and regulatory scope of their regulators. Since the regulatory relationships are inferred from pattern recognition between target-target genes, the method we present is especially suitable for inferring gene regulatory relationships in which there is a time delay between the expression of regulating and target genes. PMID- 15540197 TI - The competition between protein folding and aggregation: off-lattice minimalist model studies. AB - Protein aggregation has been associated with a number of human diseases, and is a serious problem in the manufacture of recombinant proteins. Of particular interest to the biotechnology industry is deleterious aggregation that occurs during the refolding of proteins from inclusion bodies. As a complement to experimental efforts, computer simulations of multi-chain systems have emerged as a powerful tool to investigate the competition between folding and aggregation. Here we report results from Langevin dynamics simulations of minimalist model proteins. Order parameters are developed to follow both folding and aggregation. By mapping natural units to real units, the simulations are shown to be carried out under experimentally relevant conditions. Data pertaining to the contacts formed during the association process show that multiple mechanisms for aggregation exist, but certain pathways are statistically preferred. Kinetic data show that there are multiple time scales for aggregation, although most association events take place at times much shorter than those required for folding. Last, we discuss results presented here as a basis for future work aimed at rational design of mutations to reduce aggregation propensity, as well as for development of small-molecular weight refolding enhancers. PMID- 15540198 TI - Molecular and phase toxicity of compressed and supercritical fluids in biphasic continuous cultures of Clostridium thermocellum. AB - A novel continuous high-pressure biphasic bioreactor was designed to investigate the toxicity of compressed and supercritical fluids on the thermophilic bacterium Clostridium thermocellum. Cultures were conducted at 1.8 and 7.0 MPa hydrostatic pressure and in the presence of compressed N(2) (7.0 MPa), gaseous (1.8 MPa) and supercritical ethane (7.0 MPa), and gaseous (1.8 MPa) and liquid (7.0 MPa) propane at a single dilution rate. No significant changes in metabolism or growth were observed in the presence of compressed N(2) relative to 7.0 MPa hydrostatic pressure, indicating that it acted as an inert fluid. However, dramatic inhibitions of growth and metabolism occurred in the presence of ethane and propane at 7.0 MPa. These inhibitions were reversed by depressurization from the supercritical (ethane) or liquid (propane) to gaseous state. Solvent toxicity by compressed and supercritical fluids was attributed to phase toxicity and was correlated with fluid density rather than conventional measures of toxicity (log P(o/w)). This biphasic reactor system facilitates investigations of solvent toxicity and dissolved gas effects on whole cells under elevated pressures. PMID- 15540199 TI - A biocatalyst for the removal of sulfite from alcoholic beverages. AB - The presence of sulfites in alcoholic beverages, particularly in wines, can cause allergic responses with symptoms ranging from mild gastrointestinal problems to life threatening anaphylactic shock in a substantial portion of the population. We have developed a simple and inexpensive biocatalytic method that employs wheatgrass (Triticum aestivum) chloroplasts for the efficient oxidation of sulfites in wines to innocuous sulfates. A sufficiently high rate of sulfite oxidation was obtained in the presence of ethanol at concentrations commonly found in most wines. Crude chloroplast preparations at a concentration as low as 5 mg/mL were capable of reducing sulfite in commercial white wines from 150 ppm to under 7.5 ppm within 3 hours. A 93% removal of sulfite in commercial red wines was observed with 1 mg/mL chloroplasts within 45 min. Optimal sulfite removal efficiency was observed at pH 8.5 and was promoted by illumination, indicating the participation of light-induced photosynthetic electron transport processes in sulfite oxidation. Overall, this work indicates that biocatalytic oxidation using wheatgrass chloroplasts can be employed to remove sulfites from beverages prior to consumption. PMID- 15540200 TI - Growth of Leuconostoc mesenteroides NRRL-B523 in an alkaline medium: suboptimal pH growth inhibition of a lactic acid bacterium. AB - Bacterial profile modification (BPM), a form of tertiary oil recovery, diverts water from the water-flooded high-permeability zone into the oil-bearing low permeability zone. During field use, exopolymer-producing bacteria plug the high permeability zone only in the immediate vicinity of the injection point (the near well bore region). For effective BPM the plug must penetrate far into the formation. Slowing the specific growth rate, lengthening the lag phase, and slowing the polymerization rate are techniques that can prolong the onset of biopolymer gelation and extend the depth of the biological plug. In batch experiments, the growth of Leuconostoc mesenteroides NRRL-B523 was inhibited by the synergistic effects of high substrate loading and an alkaline pH. Exponential growth was delayed up to 190 h. It was observed that cell division was significantly retarded until the medium pH, reduced by the acid byproducts of fermentation, reached a critical value of 6.79 +/- 0.06. A mathematical model was developed to describe the relationship between specific growth rate, lag time, and medium pH. PMID- 15540201 TI - Aberrant expression of serine/threonine kinase Pim-3 in hepatocellular carcinoma development and its role in the proliferation of human hepatoma cell lines. AB - Most cases of human hepatocellular carcinoma develop after persistent chronic infection with human hepatitis B virus or hepatitis C virus, and host responses are presumed to have major roles in this process. To recapitulate this process, we have developed the mouse model of hepatocellular carcinoma using hepatitis B virus surface antigen transgenic mice. To identify the genes associated with hepatocarcinogenesis in this model, we compared the gene expression patterns between pre-malignant lesions surrounded by hepatocellular carcinoma tissues and control liver tissues by using a fluorescent differential display analysis. Among the genes that were expressed differentially in the pre-malignant lesions, we focused on Pim-3, a member of a proto-oncogene Pim family, because its contribution to hepatocarcinogenesis remains unknown. Moreover, the unavailability of the nucleotide sequence of full-length human Pim-3 cDNA prompted us to clone it from the cDNA library constructed from a human hepatoma cell line, HepG2. The obtained 2,392 bp human Pim-3 cDNA encodes a predicted open reading frame consisting of 326 amino acids. Pim-3 mRNA was selectively expressed in human hepatoma cell lines, but not in normal liver tissues. Moreover, Pim-3 protein was detected in human hepatocellular carcinoma tissues and cell lines but not in normal hepatocytes. Furthermore, cell proliferation was attenuated and apoptosis was enhanced in human hepatoma cell lines by the ablation of Pim-3 gene with RNA interference. These observations suggest that aberrantly expressed Pim-3 can cause autonomous cell proliferation or prevent apoptosis in hepatoma cell lines. PMID- 15540202 TI - Endostatin inhibits nitric oxide and diminishes VEGF and collagen XVIII in squamous carcinoma cells. AB - Low pO(2) values are a common finding among oral squamous cell carcinomas (SCC). Our objective was to determine the role that oxygen tension plays on the direct tumor effect of endostatin (ES). Squamous carcinoma cell lines were grown under normoxic or hypoxic conditions and treated with endostatin (ES), nitric oxide (NO) donors, NO scavengers, NO synthase inhibitors, or transduced with AdenoVec hEndo or AdenoVec Null vectors. The expression of vascular endothelial growth factor (VEGF) and collagen XVIII were determined by RT-PCR and protein levels assessed by Western blot analyses. Our studies demonstrated that collagen XVIII and VEGF are expressed and responsive to ES in a limited number of SCC cell lines during normoxia but were most responsive when grown under hypoxic conditions. VEGF and collagen XVIII were downregulated by both ES and transduction of cells with AdenoVec-hEndo. The effects of ES on SCC cells were enhanced by aminoguanidine (Ag), L-NAME, and diphenyleneiodonium chloride (DPI). Endostatin and transduced with ES vectors diminished the levels of NO whereas NO donors enhanced VEGF expression and collagen XVIII expression. In conclusion, the direct effect of endostatins on tumor cells is most effective under conditions of low oxygen tension and can be potentiated by the use of nitric oxide synthase inhibitors or NO scavengers. PMID- 15540203 TI - Challenges in mass spectrometry-based proteomics. AB - During the last decade, protein analysis and proteomics have been established as new tools for understanding various biological problems. As the identification of proteins after classical separation techniques, such as two-dimensional gel electrophoresis, have become standard methods, new challenges arise in the field of proteomics. The development of "functional proteomics" combines functional characterization, like regulation, localization and modification, with the identification of proteins for deeper insight into cellular functions. Therefore, different mass spectrometric techniques for the analysis of post-translational modifications, such as phosphorylation and glycosylation, have been established as well as isolation and separation methods for the analysis of highly complex samples, e.g. protein complexes or cell organelles. Furthermore, quantification of protein levels within cells is becoming a focus of interest as mass spectrometric methods for relative or even absolute quantification have currently not been available. Protein or genome databases have been an essential part of protein identification up to now. Thus, de novo sequencing offers new possibilities in protein analytical studies of organisms not yet completely sequenced. The intention of this review is to provide a short overview about the current capabilities of protein analysis when addressing various biological problems. PMID- 15540204 TI - Cerebrospinal fluid protein patterns in neurodegenerative disease revealed by liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry. AB - This study demonstrates the power of a novel proteomic approach developed for the detection and identification of biological markers in body fluids. The goal was to observe alterations in the protein patterns of cerebrospinal fluid (CSF) related to amyotrophic lateral sclerosis (ALS), a neurodegenerative disorder with unknown etiology. In the experiments, tryptic digests of CSF from patients and healthy controls were analyzed by on-line capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry. (FT-ICR MS) Typically, around 4000 peptides were detected in one such experiment, and a pattern recognition program was constructed for the data analysis to distinguish mass chromatograms from patients and controls. This strategy was evaluated comparing the peptide patterns of CSF spiked in vitro with a biomarker, with control CSF. The patterns were clearly separated and the tryptic peptides of the biomarker were successfully selected as characteristic peaks. Hence, the method was applied to compare mass chromatograms of CSF from 12 ALS-patients and 10 matched healthy controls. While no biomarker alone could be identified from the characteristic peaks, we were able to assign 4 out of 5 unknown samples correctly (i.e., 80% correctly diagnosed, 20% false-negative), and it would be 100% if we reject a possible outlier believed to be caused by an occlusion in the spinal CSF compartment. These findings are very promising, although the clinical relevance is not fully established due to the low number of unknown samples analyzed. In addition to the diagnostic potential, these results may be important steps towards understanding the neurodegenerative process. PMID- 15540205 TI - CXCR4 expression reflects tumor progression and regulates motility of bladder cancer cells. AB - Transitional cell carcinoma of the urinary bladder remains life threatening due to the high occurrence of metastases. Emerging evidence suggests that chemokines and their receptors play a critical role in tumor metastases. In our study, we performed a systematic analysis of the mRNA and protein expression levels of all 18 chemokine receptors in normal urothelium and bladder cancer. CXCR4 was the only chemokine receptor whose mRNA expression level was upregulated in bladder cancer cell lines as well as in invasive and locally advanced bladder cancer tissue samples (pT2-pT4). In contrast, superficial bladder tumors (pTa and pT1) displayed low CXCR4 expression levels and normal urothelial cells were negative for CXCR4. Immunohistochemistry of a bladder cancer tissue microarray (TMA) confirmed that a subgroup of invasive bladder cancers revealed a high CXCR4 protein expression, while superficial bladder tumors showed low immunoreactivity. To investigate the functional significance of CXCR4 expression, we performed migration and invasion assays. Exposure of CXCR4-positive bladder cancer cells to CXCL12 in a Boyden chamber type assay provoked a significant increase in migration as well as invasion across a Matrigel barrier. Enhanced migration and invasion were inhibited by a CXCR4-specific blocking antibody. In contrast, normal urothelial cells did not respond to CXCL12 and lacked chemotactic migration. In conclusion, bladder cancer cells express CXCR4 progressively with advanced tumorigenesis and this receptor interacts with CXCL12 to mediate tumor chemotaxis and invasion through connective tissue. These properties identify CXCR4 as a potential target for the attenuation of bladder cancer metastases. PMID- 15540206 TI - Comparison of genomic abnormalities between BRCAX and sporadic breast cancers studied by comparative genomic hybridization. AB - Very little is known about the chromosomal regions harbouring genes involved in initiation and progression of BRCAX-associated breast cancers. We applied comparative genomic hybridization (CGH) to identify the most frequent genomic imbalances in 18 BRCAX hereditary breast cancers and compared them to chromosomal aberrations detected in a group of 27 sporadic breast cancers. The aberrations observed most frequently in BRCAX tumours were gains of 8q (83%), 19q (67%), 19p (61%), 20q (61%), 1q (56%), 17q (56%) and losses of 8p (56%), 11q (44%) and 13q (33%). The sporadic cases most frequently showed gains of 1q (67%), 8q (48%), 17q (37%), 16p (33%), 19q (33%) and losses of 11q (26%), 8p (22%) and 16q (19%). Losses of 8p and gains 8q, 19 as well as gains of 20q (with respect to ductal tumours only) were detected significantly more often in BRCAX than in sporadic breast cancers. Analysis of 8p-losses and 8q-gains showed that these aberrations are early events in the tumorigenesis of BRCAX tumors. The findings of this report indicate similarities between BRCAX and BRCA2 tumours, possibly suggesting a common pathway of disease. These findings need confirmation by more extensive studies because only a limited number of cases were analysed and there are relatively few reports published. PMID- 15540207 TI - Zooming in: fractionation strategies in proteomics. AB - The recent development of mass spectrometry, i.e., high sensitivity, automation of protein identification and some post-translational modifications (PTMs) significantly increased the number of large-scale proteomics projects. However, there are still considerable limitations as none of the currently available proteomics techniques allows the analysis of an entire proteome in a single step procedure. On the other hand, there are several successful studies analyzing well defined groups of proteins, e.g., proteins of purified organelles, membrane microdomains or isolated proteins with certain PTMs. Coupling of advanced separation methodologies (different prefractionation strategies, such as subcellular fractionation, affinity purification, fractionation of proteins and peptides according to their physicochemical properties) to highly sensitive mass spectrometers provides powerful means to detect and analyze dynamic changes of low abundant regulatory proteins in eukaryotic cells on the subcellular level. This review summarizes and discusses recent strategies in proteomics approaches where different fractionation strategies were successfully applied. PMID- 15540208 TI - Identification of proteins induced by polycyclic aromatic hydrocarbon in Mycobacterium vanbaalenii PYR-1 using two-dimensional polyacrylamide gel electrophoresis and de novo sequencing methods. AB - Protein profiles of Mycobacterium vanbaalenii PYR-1 grown in the presence of high molecular-weight polycyclic aromatic hydrocarbons (HMW PAHs) were examined by two dimensional gel electrophoresis (2-DE). Cultures of M. vanbaalenii PYR-1 were incubated with pyrene, pyrene-4,5-quinone (PQ), phenanthrene, anthracene, and fluoranthene. Soluble cellular protein fractions were analyzed and compared, using immobilized pH gradient (IPG) strips. More than 1000 gel-separated proteins were detected using a 2-DE analysis program within the window of isoelectric point (pI) 4-7 and a molecular mass range of 10-100 kDa. We observed variations in the protein composition showing the upregulation of multiple proteins for the five PAH treatments compared with the uninduced control sample. By N-terminal sequencing or mass spectrometry, we further analyzed the proteins separated by 2 DE. Due to the lack of genome sequence information for this species, protein identification provided an analytical challenge. Several PAH-induced proteins were identified including a catalase-peroxidase, a putative monooxygenase, a dioxygenase small subunit, a small subunit of naphthalene-inducible dioxygenase, and aldehyde dehydrogenase. We also identified proteins related to carbohydrate metabolism (enolase, 6-phosphogluconate dehydrogenase, indole-3-glycerol phosphate synthase, and fumarase), DNA translation (probable elongation factor Tsf), heat shock proteins, and energy production (ATP synthase). Many proteins from M. vanbaalenii PYR-1 showed similarity with protein sequences from M. tuberculosis and M. leprae. Some proteins were detected uniquely upon exposure to a specific PAH whereas others were common to more than one PAH, which indicates that induction triggers not only specific responses but a common response in this strain. PMID- 15540209 TI - Quantitative protein profiling using antibody arrays. AB - Traditional approaches to microarrays rely on direct binding assays where the extent of hybridisation and the signal detected are a measure of the analyte concentration in the experimental sample. This approach, directly imported from the nucleic acid field, may fail if applied to antibody-antigen interactions due to the shortage of characterised antibodies, the significant heterogeneity of antibody affinities, their dependence on the extent of protein modification during labelling and the inherent antibody cross-reactivity. These problems can potentially limit the multiplexing capabilities of protein affinity assays and in many cases rule out quantitative protein profiling using antibody microarrays. A number of approaches aimed at achieving quantitative protein profiling in a multiplex format have been reported recently. Of those reported, the three most promising routes include signal amplification, multicolour detection and competitive displacement approaches to multiplex affinity assays. One in particular, competitive displacement, also overcomes the problems associated with quantitation of affinity interactions and provides the most generic approach to highly parallel affinity assays, including antibody arrays. PMID- 15540210 TI - Hypermethylation of RASSF1A and BLU tumor suppressor genes in non-small cell lung cancer: implications for tobacco smoking during adolescence. AB - The putative tumor suppressors RASSF1A and BLU are mapped adjacent to one another on chromosome 3p21.3, a region frequently deleted in lung cancer. These genes are often inactivated by promoter hypermethylation, but the association of this inactivation with clinical features of the disease or with carcinogen exposure has been poorly studied. Early age starting smoking has been hypothesized as an independent risk factor for lung cancer, and mechanistically, adolescence may constitute a critical period for tobacco carcinogen exposure. To study the relationship of tobacco smoke exposure with hypermethylation of RASSF1A and BLU, methylation-specific PCR was performed on a case series study of incident, surgically resected non-small cell lung cancer (NSCLC), and the prevalence of this alteration was examined in relation to clinical and exposure information collected on the patients. Hypermethylation of the RASSF1A promoter occurred in 47% (83/178) and of the BLU promoter in 43% (68/160) of NSCLC tumors examined. There was no significant association between methylation of these 2 genes, but methylation of either of these genes tended to occur more often in the adenocarcinoma (AC) histology compared to squamous cell carcinoma (SCC). Controlling for pack-years smoked, age, gender and histology, starting smoking under age 18 was significantly related to RASSF1A methylation [prevalence ratio (PR) = 1.6, 95% confidence interval [CI] = 1.1-2.3]. These results indicate that starting smoking under age 18 is an independent risk for RASSF1A hypermethylation, thus identifying a molecular alteration related to the epidemiologic effect of teenage smoking as a lung cancer risk. PMID- 15540211 TI - Rapid enrichment of human papillomavirus (HPV)-specific polyclonal T cell populations for adoptive immunotherapy of cervical cancer. AB - The majority of cervical cancers are caused by human papillomavirus type 16 (HPV16). Cervical cancer is associated with an ineffective host immune response against the HPV16 oncoproteins, characterized by the lack of the strong E6 specific T-helper type 1 (Th1) immunity that is generally present in healthy individuals, the presence of improperly polarized HPV16E6- and E7-specific CD4(+) T cells and increased numbers of regulatory T cells. Therefore, immunotherapeutic intervention is likely to require a modality that deletes the regulatory T cell component and enhances the HPV16-specific Type 1 T cell response. HLA-matched allogeneic stem cell transplantation may offer such a modality, because it involves the eradication of host immune cells and enables the transfer of donor derived tumor-specific T cells to the patient. As a first step in the development of such a treatment, we evaluated the success rate of a protocol for enrichment of HPV16E6-specific CD4(+) T cells from healthy donor PBMC on the basis of their IFNgamma secretion. After a short in vitro stimulation with overlapping 30 amino acid long HPV16E6 peptides, we enriched the IFNgamma secreting cells by magnetic cell sorting. The obtained polyclonal CD4(+) T cell populations recognized distinct epitopes within HPV16E6, as well as E6 protein, processed and presented by autologous professional antigen presenting cells. The described protocol proved successful in PBMC from more than half of the healthy adult blood donors. These HPV16E6-specific CD4(+) T cells may turn out to be an essential component of future adoptive T cell therapy for advanced cervical cancer, by orchestrating CTL dependent and independent tumoricidal mechanisms. PMID- 15540212 TI - Towards a comprehensive understanding of Bacillus subtilis cell physiology by physiological proteomics. AB - Using Bacillus subtilis as a model system for functional genomics, this review will provide insights how proteomics can be used to bring the virtual life of genes to the real life of proteins. Physiological proteomics will generate a new and broad understanding of cellular physiology because the majority of proteins synthesized in the cell can be visualized. From a physiological point of view two major proteome fractions can be distinguished: proteomes of growing cells and proteomes of nongrowing cells. In the main analytical window almost 50% of the vegetative proteome expressed in growing cells of B. subtilis were identified. This proteomic view of growing cells can be employed for analyzing the regulation of entire metabolic pathways and thus opens the chance for a comprehensive understanding of metabolism and growth processes of bacteria. Proteomics, on the other hand, is also a useful tool for analyzing the adaptational network of nongrowing cells that consists of several partially overlapping regulation groups induced by stress/starvation stimuli. Furthermore, proteomic signatures for environmental stimuli can not only be applied to predict the physiological state of cells, but also offer various industrial applications from fermentation monitoring up to the analysis of the mode of action of drugs. Even if DNA array technologies currently provide a better overview of the gene expression profile than proteome approaches, the latter address biological problems in which they can not be replaced by mRNA profiling procedures. This proteomics of the second generation is a powerful tool for analyzing global control of protein stability, the protein interaction network, protein secretion or post-translational modifications of proteins on the way towards the elucidation of the mystery of life. PMID- 15540213 TI - Vascular proteomics: linking proteomic and metabolomic changes. AB - Cardiovascular diseases constitute the largest of death in the Western world. Various stressors, including elevated blood pressure, smoking, diabetes, and hypercholesterolemia directly or indirectly damage the vessel wall, eventually inducing arterial stiffness (arteriosclerosis) and lipid accumulation (atherosclerosis). However, the molecular mechanisms of atheroma formation are not yet fully clarified. While many investigators have used proteomic techniques to study cardiac diseases, vascular proteomics is still in its infancy. The present review highlights studies, in which proteomics has been successfully applied to study protein alterations in the vasculature. Furthermore, we will summarize our recent progress in combining proteomic and metabolomic techniques to reveal protein and metabolite alterations in the cardiovascular system: two dimensional (2-D) gel electrophoresis proved to be highly complementary to nuclear magnetic resonance (NMR) spectroscopy, in that post-translational modifications of the most abundant enzymes were displayed on 2-D gels while NMR spectroscopy revealed changes in the corresponding metabolites. Importantly, the simultaneous assessment of protein and metabolite changes translated purely descriptive proteomic and metabolomic profiles into a functional context and provided important insights into pathophysiological mechanisms that would not have been obtained by other techniques. PMID- 15540214 TI - Active and passive smoking and breast cancer risk in middle-aged Japanese women. AB - To examine the hypothesis that tobacco smoke is associated with the risk of female breast cancer, we estimated the relative risks of active and passive smoke in middle-aged Japanese women in a population-based prospective study. The cohort consisted of residents in 4 public health center areas, aged 40 to 59 years. A self-administered questionnaire survey was conducted in 1990. This analysis included 21,805 subjects, 180 of whom had developed breast cancer by December 31, 1999. When the reference was defined as never-active smokers without passive smoking, adjusted relative risks (RRs) were 1.9 (95% confidence interval [CI] = 1.0-3.6) in current active smokers, 1.2 (95% CI = 0.4-4.0) in ex-active smokers and 1.2 (95% CI = 0.8-1.6) in never-active smokers with passive smoking. The elevated risk for ever-smokers was clearly observed in premenopausal women at baseline (RR = 3.9, 95% CI = 1.5-9.9) but not in postmenopausal women (RR = 1.1, 95% CI = 0.5-2.5). In never-active smokers, the adjusted RR for passive smoking, residential or occupational/public tobacco smoke exposure was 1.1 (95% CI = 0.8 1.6). In premenopausal women, passive smoking increased the risk (RR = 2.6; 95% CI = 1.3-5.2) but not in postmenopausal women (RR = 0.7; 95% CI = 0.4-1.0). We conclude that tobacco smoking increases the risk of female breast cancer in premenopausal women. PMID- 15540215 TI - Microarray-based gene expression profiling of benign, atypical and anaplastic meningiomas identifies novel genes associated with meningioma progression. AB - To identify gene expression profiles associated with human meningiomas of different World Health Organization (WHO) malignancy grades, we analyzed 30 tumors (13 benign meningiomas, WHO grade I; 12 atypical meningiomas, WHO grade II; 5 anaplastic meningiomas, WHO grade III) for the expression of 2,600 genes using cDNA-microarray technology. Receiver operator curve (ROC) analysis with a cutoff value of 45% selection probability identified 37 genes with decreased and 27 genes with increased expression in atypical and anaplastic meningiomas, compared to benign meningiomas. Supervised classification of the tumors did not reveal specific expression patterns representative of each WHO grade. However, anaplastic meningiomas could be distinguished from benign meningiomas by differential expression of a distinct set of genes, including several ones associated with cell cycle regulation and proliferation. Investigation of potential correlations between microarray expression data and genomic aberrations, detected by comparative genomic hybridization (CGH), demonstrated that losses on chromosomes 10 and 14 were associated with distinct expression profiles, including increased expression of several genes related to the insulin like growth factor (IGF) (IGF2, IGFBP3 and AKT3) or wingless (WNT) (CTNNB1, CDK5R1, ENC1 and CCND1) pathways. Taken together, our microarray-based expression profiling revealed interesting novel candidate genes and pathways that may contribute to meningioma progression. PMID- 15540216 TI - Is it functional? Report on the first BSPR/EBI meeting on functional proteomics. AB - This report describes the first scientific meeting of the British Society for Proteome Research (BSPR), which was organised jointly with the European Bioinformatics Institute (EBI) and held in July 2004. The focus of the conference was functional proteomics with an emphasis on possible clinical application. The main subjects described here are: the need to simplify samples, the use of biological fluids versus tissue, consideration of biological and experimental variation and the creation of databases to achieve meaningful functional analysis. PMID- 15540217 TI - MYST3/CREBBP (MOZ/CBP) and CREBBP/MYST3 (CBP/MOZ) transcripts in AML with t(8;16)(p11;p13). PMID- 15540218 TI - Frequent microsatellite instability in primary esophageal carcinoma associated with extraesophageal primary carcinoma. AB - Patients with esophageal squamous cell carcinoma (ESCC) frequently develop other primary cancers, such as gastric cancer and head and neck cancer. Details of carcinogenesis in patients with multiple primaries that include esophageal carcinoma with other primary carcinoma (ECOPC) remain uncertain. We examined microsatellite instability (MSI) status, frameshift mutation in target genes of MSI, mismatch repair protein expression and hypermethylation of the hMLH1 promoter region in ECOPC patients to better understand the underlying carcinogenic processes. High frequency MSI (MSI-H) was found in 15 (44.1%) of 34 patients with ECOPC, but in only 6 (14.3%) of 42 patients with esophageal cancer alone (p < 0.01). Frameshift mutations in TGFbetaRII, BAX, MSH3 and MSH6 genes respectively were present in 4, 1, 2 and 2 of 34 ECOPC patients. Immunohistochemical study showed that 12 (80.0%) of 15 MSI-H tumors showed loss of expression of either hMLH1 or hMSH2. In addition, 6 of 9 tumors (66.7%) that showed reduced hMLH1 expression also had hypermethylation of the hMLH1 promoter region. Our findings suggested that carcinogenesis in ECOPC was closely associated with the MSI pathway because of mismatch repair protein deficiency. PMID- 15540219 TI - A novel dipeptide-based HIV protease inhibitor containing allophenylnorstatine. AB - Dipeptide analogues incorporating allophenylnorstatine [Apns; (2S,3S)-3amino-2 hydroxy-4-phenylbutyric acid] as a transition state mimic at the scissile bond were designed and synthesized in the hope of obtaining a novel KNI series of HIV protease inhibitors. The precursors, N-P2'-3-(2S,3S)-3-(tert-butyloxy carbonyl)amino-2-hydroxy-4-phenylbutanoyl)-5,5-dimethylthiazolidine-4-carboxamide (N-Boc-Apns-Dmt-P2') 4a-p were prepared by deprotection of the synthones N-P2' (tert-butyloxycarbonyl)-5,5-dimethylthiazolidine-4-carboxamide (Boc-Dmt-P2') 2a p, then coupling with (2S,3S)3-(tert-butyloxycarbonyl)amino-2-hydroxy-4 phenylbutanoic acid (N-Boc-Apns-OH) 3. The deprotected intermediates 4 were coupled with the activated carboxyl groups of the P2 ligands to afford the target dipeptides. In this work, we fixed at the P2 site either a 2,6 dimethylphenoxyacetyl or a 3-hydroxy-2-methylbenzoyl group. Substitutes at the P2' site were varied to afford the members of the series 7 and 8. Improved activity of most of the members of series 8 relative to their analogues of series 7 can be partially attributed to the differences in the structures of the P2 moieties. Positional isomerism in the P2' moieties significantly affected the activity and polarity of the target. PMID- 15540220 TI - A new family of synthetic diterpenes that regulates cytokine synthesis by inhibiting IkappaBalpha phosphorylation. AB - The synthesis and the biological evaluation of a new family diterpenes are presented. The synthetic studies were inspired by the structural framework of acanthoic acid (1) and yielded a family of compounds that were evaluated as anti inflammatory agents. Among them, compounds 2, 10, 12, and 16 exhibited a very low nonspecific cytotoxicity and inhibited the synthesis of TNF-alpha with greater than 65 % efficacy at low micromolar concentrations. Cytokine-specificity studies revealed that these compounds also inhibited the synthesis of the proinflammatory cytokines IL-1beta and IL-6, while inhibition of IL-1ra and IL-8 synthesis was marginal and only occurred at high concentrations. Further studies, through EMSA and Western blot analyses, indicated that these compounds decreased the extent of phosphorylation of IkappaBalpha; this suggests that they exert their anti inflammatory profile by inhibiting NF-kappaB-mediated cytokine synthesis. These findings imply that these diterpenes represent promising leads for the development of novel anti-inflammatory agents. PMID- 15540221 TI - The role of nutrition in cancer development and prevention. AB - Nutrition has long been suspected to play an important role in cancer etiology. The biologic properties of nutrients make them prime candidates to aid in cancer prevention. Indeed, early epidemiologic cancer studies seemed to confirm the relevance of diet. These studies, however, were plagued by recall bias and confounding and may have thus been misleading. The results from large, prospective cohorts do not support a strong relation between nutrition and cancer. Given the difficulties in precisely assessing habitual diet, modest associations may exist, which may be impossible to capture when relying on self reported dietary information. Energy balance, reflected in a low body weight and high level of physical activity, has been more convincingly related to lower cancer rates. The potential importance of more extreme dietary regimens and of nutrition during earlier periods of life remains to be explored. Finally, epigenetic research is likely to contribute to the understanding of nutritional regulation of gene expression. PMID- 15540222 TI - Activation of HLXB9 by juxtaposition with MYB via formation of t(6;7)(q23;q36) in an AML-M4 cell line (GDM-1). AB - Mutation or dysregulation of related homeobox genes occurs in leukemia. Using RT PCR, we screened members of the EHG family of homeobox genes, comprising EN1 (at 2q14), GBX2 (at 2q36), and EN2, GBX1, and HLXB9 (at 7q36), for dysregulation in acute myeloid leukemia (AML) cell lines indicated by chromosomal breakpoints at these sites. Only one EHG-family gene was expressed, HLXB9, in cell line GDM-1 (AML-M4). Karyotypic analysis of GDM-1 revealed a unique t(6;7)(q23;q35), also present in the patient. Fluorescence in situ hybridization analysis showed chromosomal breakpoints close to the region upstream of HLXB9, at 7q36, a region rearranged in certain AML patients, and at 6q23 upstream of MYB, a gene activated in leukemia. Detailed expression analysis suggested ectopic activation of HLXB9 occurred via juxtaposition with regions upstream of MYB, which was highly expressed in GDM-1. Our data identified a cell line model for a novel leukemic translocation involving MYB with HLXB9, further implicating HLXB9 in leukemogenesis. PMID- 15540223 TI - Synthesis and biological evaluation of novel 2',3',4'-triply branched carbocyclic nucleosides as potential antiviral agents. AB - Novel 2',3',4'-triply branched carbocyclic nucleosides were synthesized in this study. The introduction of two methyl groups in the 2'- and 3'-position was accomplished by a Horner-Wadsworth-Emmons reaction and isopropenyl magnesium bromide addition, respectively. The construction of the required 4'-quaternary carbon was carried out using a [3,3]-sigmatropic rearrangement. Bis-vinyls were successfully cyclized using a Grubbs' catalyst II. The natural bases (adenine, cytosine) were efficiently coupled using a Pd(0) catalyst. The antiviral activities of the synthesized compounds were evaluated against HIV-1, HSV-1, HSV 2 and HCMV. Compound 30 displayed moderate anti-HCMV activity (EC50 = 30.1 microg/mL), without exhibiting any cytotoxicity at up to 100 microM. PMID- 15540224 TI - Role of the polymorphic IL-1B, IL-1RN and TNF-A genes in distal gastric cancer in Mexico. AB - Several cytokine gene polymorphisms have been associated with increased risk of distal gastric cancer (GC) and its precursor histological markers in Caucasian, Asian and Portuguese populations although little is known about their role in other ethnic groups. Our study investigates the role of the IL-1B-31, IL-1RN and TNF-A-308 gene polymorphisms as risk factors for the development of GC in a Mexican population. We studied 278 patients who were enrolled at the Hospital Universitario Dr. Jose Eleuterio Gonzalez, Universidad Autonoma de Nuevo Leon. The subjects were divided into 2 groups. Sixty-three patients with histologically confirmed distal GC (mean age = 58.8 years, range = 22-84, F:M = 0.56), and 215 patients with no evidence of distal or proximal GC (mean age = 56.1 years, range = 18-92, F:M = 1.17). The IL-1B-31 and the TNF-A-308 polymorphisms were determined by PCR-RFLP and pyrosequencing, respectively, in all cases and controls. The VNTR polymorphism in intron 2 of the 1L-1RN gene was typed by PCR in 25 cases and 201 controls. The H. pylori status was determined by histology, rapid urease test, culture and serology for non-cancer controls and by histology for the GC cases. The carriage of the proinflammatory IL-1B-31*C allele was associated with increased risk of distal GC (odds ratio [OR] = 7.63, 95% confidence interval [CI] = 1.73-46.94, p = 0.003). When cases and controls were matched by age and gender, the OR value was higher (OR = 8.05, 95% CI = 1.8 50.22, p = 0.001). When only H. pylori GC cases and controls were compared, the OR value was 7.8 (95% CI = 1.05-161.8, p = 0.04). No association was found between any of the other polymorphisms studied and distal GC. In this Mexican population, the IL-1B proinflammatory genotype increases the risk of distal GC. These findings are similar to previous reports in Caucasian populations and underscore the importance of cytokine gene polymorphisms in the development of distal GC. PMID- 15540225 TI - Postmenopausal levels of sex hormones and risk of breast carcinoma in situ: results of a prospective study. AB - We report on a prospective study to assess the association of postmenopausal serum levels of sex hormones with subsequent risk of breast carcinoma in situ. We conducted a case-control study nested within the cohort of the New York University Women's Health Study, a large prospective study documenting a positive association of circulating levels of estrogens and androgens with invasive breast cancer. The study included 69 cases of incident in situ carcinoma and 134 individually matched controls. No statistically significant trend of increasing risk with increasing level of any of the hormones was observed. Odds ratios (95% CIs) for the highest tertile relative to the lowest were 1.10 (0.51-2.39) for estradiol, 0.95 (0.41-2.19) for estrone, 1.63 (0.69-3.88) for testosterone, 0.99 (0.44-2.24) for androstenedione, 0.99 (0.45-2.20) for dehydroepiandrosterone sulfate and 0.81 (0.38-1.74) for sex hormone-binding globulin. Adjusting for potential confounders did not materially affect the results, nor did limiting the analysis to the 59 cases of ductal carcinoma in situ, the lesion thought to be the direct precursor of most invasive breast cancers. Our results are at variance with the positive associations observed in this same cohort with risk of invasive breast cancer. Possible explanations for our results include lack of power, an effect of sex hormones limited to the progression from in situ to invasive tumors, overrepresentation of indolent tumors or an effect of sex hormones on the induction of only a subset of in situ tumors, those that would develop into invasive tumors. PMID- 15540226 TI - Synthesis of dimeric trifluoromethoxyacridine-derived pathogen-inactivating nucleic acid intercalators. AB - A series of antiviral active compounds consisting of an intercalating acridine derived part, a spacer region and a reactive EDTA-derived conjugate was synthesized in an easy sequence. Thus, suitably mono-protected 1,omega alkyldiamines gave, upon reaction with 9-chloro-2-trifluoromethoxyacridine, followed by deprotection and reaction with EDTA dianhydride, the target molecules. Incorporation of their Fe(II) complexes in the presence of ascorbate gave a reduction of the phage titer of MS2 phages by several logarithmic decades. PMID- 15540227 TI - Oligonucleotide bearing ethylenediamine-N,N,N'-Triacetates for gap-selective DNA hydrolysis by Ce4+/EDTA. AB - With the use of two oligonucleotides bearing ethylenediamine-N,N,N'-triacetate groups as additives, gap sites were formed at predetermined sites in substrate DNA. Upon treating these systems with a Ce(4+)/EDTA complex at pH 7.0 and 37 degrees C, the phosphodiester linkages at the gap site were selectively hydrolyzed. The DNA scission was greatly promoted by the introduction of ethylenediaminetriacetate groups, and the scission efficiency increased as the number of these groups increased. Even a one-base gap was successfully hydrolyzed when three ethylenediaminetriacetate groups were placed consecutively at both edges of the gap, although the scission was minimal in the absence of these groups. The site-selective scission could be also achieved at higher temperatures without any significant loss of site-selectivity. PMID- 15540228 TI - Dominant B cell epitope from NY-ESO-1 recognized by sera from a wide spectrum of cancer patients: implications as a potential biomarker. AB - Monitoring the spontaneous antibody (Ab) response against a panel of relevant tumor-associated antigens (TAA) in cancer patients may provide useful information regarding the clinical status of cancer. However, current Ab detection approaches require the purification of recombinant proteins, which is often difficult to achieve. In order to bypass the purification of recombinant proteins, we identified a dominant B-cell epitope from a shared tumor antigen NY-ESO-1. A synthetic peptide of the epitope, ESO:1-40, was as sensitive as the recombinant protein for detecting Ab against NY-ESO-1 in most patients. NY-ESO-1 specific Ab present in the sera of patients with melanoma, prostate cancer, nonsmall cell lung cancer, esophageal cancer, gastric cancer and hepatocellular carcinoma reacted with the dominant peptide at a similar frequency as the recombinant protein. To our knowledge, ESO:1-40 is the first peptide epitope recognized by sera from a wide spectrum of cancer patients but not healthy donors. This simple and straightforward approach may allow the investigation of the clinical significance of spontaneous Ab responses against multiple TAA and their correlation with the clinical course of malignant diseases in the future. PMID- 15540229 TI - Canonical transient receptor potential channel 4 (TRPC4) co-localizes with the scaffolding protein ZO-1 in human fetal astrocytes in culture. AB - Members of the mammalian transient receptor potential (TRP) family form cation permeable channels at the plasma membrane implicated in capacitative calcium influx after activation by either second-messenger-mediated pathways or store depletion, or both. This study shows that with the use of RT-PCR, Western blotting, and immunohistochemistry, resting astrocytes express TRPC4 at the cell membrane, particularly at sites of cell-to-cell contact. By confocal imaging and immunoelectron microscopy, we detected co-localization of TRPC4 with the scaffolding protein zonula occludens 1 (ZO-1), and demonstrated that immunoprecipitation with antibodies to ZO-1 brought down TRPC4, and vice-versa. It has been proposed that the targeting of TRPC4 to the cell membrane is dependent on the interaction of the C-terminal TRL motif with PDZ domains. Using transfection of astrocytes with myc-tagged TRPC4 or TRL-motif truncated TRPC4 (deltaTRL), we found that deltaTRL localized predominantly to a juxtanuclear compartment, whereas the wild-type protein showed cell surface distribution. Deletion of the TRL motif also reduced plasma membrane expression as assessed by cell surface biotinylation experiments. Using GST fusion proteins, we found that TRPC4 interacted with the PDZ1 domain of ZO-1 and that this was also dependent on the TRL motif. Thus, our data demonstrate that the PDZ-interacting domain of TRPC4 controls its cell surface localization. These data implicate TRPC4 in the regulation of calcium homeostasis in astrocytes, particularly as part of a signaling complex that forms at junctional sites between astrocytes. PMID- 15540231 TI - Gliosis and brain remodeling after treatment of stroke in rats with marrow stromal cells. AB - The long-term (4-month) responses to treatment of stroke in the older adult rat, using rat bone marrow stromal cells (MSCs), have not been investigated. Retired breeder rats were subjected to middle cerebral artery occlusion (MCAo) alone, or injected intravenously with 3 x 10(6) MSCs, at 7 days after MCAo. Functional recovery was measured using an adhesive-removal patch test and a modified neurological severity score. Bromodeoxyuridine, a cell proliferation marker, was injected daily for 14 before sacrifice. Animals were sacrificed 4 months after stroke. Double immunostaining was used to identify cell proliferation and cell types for axons, astrocytes, microglia, and oligodendrocytes. MSC treatment induced significant improvement in neurological outcome after MCAo compared with control rats. MSC treatment reduced the thickness of the scar wall (P < 0.05) and reduced the numbers of microglia/macrophages within the scar wall (P < 0.01). Double staining showed increased expression of an axonal marker (GAP-43), among reactive astrocytes in the scar boundary zone and in the subventricular zone in the treated rats. Bromodeoxyuridine in cells preferentially colocalized with markers of astrocytes (GFAP) and oligodendrocytes (RIP) in the ipsilateral hemisphere, and gliogenesis was enhanced in the subventricular zone of the rats treated with MSCs. This is the first report to show that MSCs injected at 7 days after stroke improve long-term neurological outcome in older animals. Brain tissue repair is an ongoing process with reactive gliosis, which persists for at least 4 months after stroke. Reactive astrocytes responding to MSC treatment of ischemia may also promote axonal regeneration during long-term recovery. PMID- 15540232 TI - State of the Art Symposium: prescreening and rescreening. AB - The effectiveness of prescreening and rescreening cervical cytology preparations whether used as quality assurance measures or in the attempt to improve sensitivity-has engendered broad discussion within the cytology community. This paper forms an introduction to some of the issues surrounding prescreening and rescreening in cervical cytology. PMID- 15540233 TI - Long-term results of high-dose rate intracavitary brachytherapy for squamous cell carcinoma of the uterine cervix. AB - BACKGROUND: The authors performed a long-term follow-up study to evaluate the efficacy and late toxicity of high-dose rate intracavitary brachytherapy (HDR ICBT) for cervical carcinoma. METHODS: From 1968 to 1986, 1148 patients with Stage IB to IVB squamous cell carcinoma of the cervix (staging was performed according to the International Federation of Gynecology and Obstetrics) were treated with a combination of external beam radiotherapy (EBRT) and HDR-ICBT. For patients with early-stage disease, 20 gray (Gy) of EBRT was delivered to the whole pelvis, followed by 24 Gy/4 fractions of HDR-ICBT and 30 Gy of central shielding EBRT. For patients with advanced-stage disease, 20-40 Gy of whole pelvic EBRT was administered, followed by 24 Gy/4 fractions of ICBT and 30-10 Gy of central-shielding EBRT. The overall treatment time was approximately 6 weeks. Among survivors, the follow-up rate was 98% and the median follow-up duration was 22 years. RESULTS: The 10-year pelvic tumor control rates were 93% for patients with Stage IB disease, 82% for patients with Stage II disease, and 75% for patients with Stage III disease. The 10-year overall and cause-specific survival rates were 74% and 89% for patients with Stage IB disease, 52% and 74% for patients with Stage II disease, and 42% and 59% for patients with Stage III disease, respectively. The 10-year actuarial rates of major complications were 4.4% in the rectosigmoid colon, 0.9% in the bladder, and 3.3% in the small intestines. CONCLUSIONS: The results of the current study suggest that the combination of EBRT and HDR-ICBT according to the authors' protocol provided outcomes that were comparable to those of the conventional low-dose rate brachytherapy with acceptable rates of late complications in the treatment of cervical carcinoma. PMID- 15540234 TI - Familial risk and clustering of nasopharyngeal carcinoma in Guangdong, China. PMID- 15540236 TI - Homocoenzyme B12 and bishomocoenzyme B12: covalent structural mimics for homolyzed, enzyme-bound coenzyme B12. AB - Efficient electrochemical syntheses of "homocoenzyme B(12)" (2, Co(beta)-(5' deoxy-5'-adenosyl-methyl)-cob(III)alamin) and "bishomocoenzyme B(12)" (3, Co(beta)-[2-(5'-deoxy-5'-adenosyl)-ethyl]-cob(III)alamin) are reported here. These syntheses have provided crystalline samples of 2 and 3 in 94 and 77 % yield, respectively. In addition, in-depth investigations of the structures of 2 and 3 in solution were carried out and a high-resolution crystal structure of 2 was obtained. The two homologues of coenzyme B(12) (2 and 3) are suggested to function as covalent structural mimics of the hypothetical enzyme-bound "activated" (that is, "stretched" or even homolytically cleaved) states of the B(12) cofactor. From crude molecular models, the crucial distances from the corrin-bound cobalt center to the C5' atom of the (homo)adenosine moieties in 2 and 3 were estimated to be about 3.0 and 4.4 A, respectively. These values are roughly the same as those found in the two "activated" forms of coenzyme B(12) in the crystal structure of glutamate mutase. Indeed, in the crystal structure of 2, the cobalt center was observed to be at a distance of 2.99 A from the C5' atom of the homoadenosine moiety and the latter was found to be present in the unusual syn conformation. In solution, the organometallic moieties of 2 and 3 were shown to be rather flexible and to be considerably more dynamic than the equivalent group in coenzyme B(12). The homoadenosine moiety of 2 was indicated to occur in both the syn and the anti conformations. PMID- 15540238 TI - Acceptance of tamoxifen chemoprevention by physicians and women at risk. PMID- 15540237 TI - Template-based recognition of protein fold within the midnight and twilight zones of protein sequence similarity. AB - Most homologous pairs of proteins have no significant sequence similarity to each other and are not identified by direct sequence comparison or profile-based strategies. However, multiple sequence alignments of low similarity homologues typically reveal a limited number of positions that are well conserved despite diversity of function. It may be inferred that conservation at most of these positions is the result of the importance of the contribution of these amino acids to the folding and stability of the protein. As such, these amino acids and their relative positions may define a structural signature. We demonstrate that extraction of this fold template provides the basis for the sequence database to be searched for patterns consistent with the fold, enabling identification of homologs that are not recognized by global sequence analysis. The fold template method was developed to address the need for a tool that could comprehensively search the midnight and twilight zones of protein sequence similarity without reliance on global statistical significance. Manual implementations of the fold template method were performed on three folds--immunoglobulin, c-lectin and TIM barrel. Following proof of concept of the template method, an automated version of the approach was developed. This automated fold template method was used to develop fold templates for 10 of the more populated folds in the SCOP database. The fold template method developed three-dimensional structural motifs or signatures that were able to return a diverse collection of proteins, while maintaining a low false positive rate. Although the results of the manual fold template method were more comprehensive than the automated fold template method, the diversity of the results from the automated fold template method surpassed those of current methods that rely on statistical significance to infer evolutionary relationships among divergent proteins. PMID- 15540239 TI - Cervical carcinoma in the elderly: an analysis of patterns of care and outcome. AB - BACKGROUND: Advanced age often is considered a poor prognostic factor for cervical carcinoma. The authors investigated the patterns of care and treatment outcomes of elderly women with cervical carcinoma. METHODS: A hospital-based tumor registry was used to identify patients with invasive cervical carcinoma who were treated between 1986 and 2003. Patients were divided into 2 cohorts: women age < 70 years and women age > or = 70 years. Survival was examined using the Kaplan-Meier method. Single and multivariate Cox proportional hazards modeling was used to estimate hazard ratios with 95% confidence intervals (95% CI). RESULTS: In total, 1582 patients were identified, including 1385 patients age < 70 years and 197 patients age > or = 70 years. The elderly patients presented with more advanced stage tumors at diagnosis (P <0.0001) and were more likely to have nonsquamous neoplasms (P=0.002). A marked difference in treatment was noted for the elderly cohort, even after stratifying by disease stage. Only 16% of the older patients underwent surgical treatment compared with 54% of the younger patients (P <0.0001). Elderly women were 9 times more likely to receive no treatment (P <0.0001). In a multivariate model of known prognostic factors, the hazard ratio for death from any cause in women age > 70 years was 2.1 (95% CI, 1.5-3.0). The hazard ratio for death from cervical carcinoma in the elderly women was 1.6 (95% CI, 1.1-2.5). CONCLUSIONS: Age is an important factor in the allocation of treatment and survival for patients with cervical carcinoma. Elderly women with cervical carcinoma are more likely to receive primary radiotherapy, to forego treatment, and to die from their disease. PMID- 15540240 TI - Aryl cation and carbene intermediates in the photodehalogenation of chlorophenols. AB - The photochemistry of 2,6-dimethyl-4-chlorophenol (6) has been studied in methanol and trifluoroethanol (TFE) through product studies and transient absorption spectroscopy. Chloride loss from triplet 6 gave triplet hydroxyphenyl cation 14, which equilibrated with triplet oxocyclohexadienylydene 15 within a few tens of nanoseconds; the cation can, however, be selectively trapped by allyltrimethylsilane (k(ad) = 10(8)-10(9) m(-1) s(-1)) to give a phenonium ion and the allylated phenol. In neat alcohols, 14 and 15 are reduced through different mechanisms, namely by hydrogen transfer through radical cation 17 and via phenoxyl radical 16, respectively. The mechanistic rationalization has been substantiated by the parallel study of an O-silylated derivative. The work shows that the chemistry of the highly (but selectively) reactive phenyl cation 14 can not only be discriminated from that of the likewise highly reactive carbene 15, but also exploited for synthetically useful reactions, as in this case with alkenes. Photolysis of electron-donating substituted halobenzenes may be the method of choice for the mild generation of some classes of phenyl cations. PMID- 15540241 TI - On the gas-phase reactivity of complexed OH+ with halogenated alkanes. AB - OH(+) is an extraordinarily strong oxidant. Complexed forms (L--OH(+)), such as H(2)OOH(+), H(3)NOH(+), or iron-porphyrin-OH(+) are the anticipated oxidants in many chemical reactions. While these molecules are typically not stable in solution, their isolation can be achieved in the gas phase. We report a systematic survey of the influence on L on the reactivity of L--OH(+) towards alkanes and halogenated alkanes, showing the tremendous influence of L on the reactivity of L--OH(+). With the help of with quantum chemical calculations, detailed mechanistic insights on these very general reactions are gained. The gas phase pseudo-first-order reaction rates of H(2)OOH(+), H(3)NOH(+), and protonated 4-picoline-N-oxide towards isobutane and different halogenated alkanes C(n)H(2n+1)Cl (n=1-4), HCF(3), CF(4), and CF(2)Cl(2) have been determined by means of Fourier transform ion cyclotron resonance measurements. Reaction rates for H(2)OOH(+) are generally fast (7.2x10(-10)-3.0x10(-9) cm(3) mol(-1) s(-1)) and only in the cases HCF(3) and CF(4) no reactivity is observed. In contrast to this H(3)NOH(+) only reacts with tC(4)H(9)Cl (k(obs)=9.2x10(-10)), while 4-CH(3) C(5)H(4)N-OH(+) is completely unreactive. While H(2)OOH(+) oxidizes alkanes by an initial hydride abstraction upon formation of a carbocation, it reacts with halogenated alkanes at the chlorine atom. Two mechanistic scenarios, namely oxidation at the halogen atom or proton transfer are found. Accurate proton affinities for HOOH, NH(2)OH, a series of alkanes C(n)H(2n+2) (n=1-4), and halogenated alkanes C(n)H(2n+1)Cl (n=1-4), HCF(3), CF(4), and CF(2)Cl(2), were calculated by using the G3 method and are in excellent agreement with experimental values, where available. The G3 enthalpies of reaction are also consistent with the observed products. The tendency for oxidation of alkanes by hydride abstraction is expressed in terms of G3 hydride affinities of the corresponding cationic products C(n)H(2n+1) (+) (n=1-4) and C(n)H(2n)Cl(+) (n=1 4). The hypersurface for the reaction of H(2)OOH(+) with CH(3)Cl and C(2)H(5)Cl was calculated at the B3 LYP, MP2, and G3(m*) level, underlining the three mechanistic scenarios in which the reaction is either induced by oxidation at the hydrogen or the halogen atom, or by proton transfer. PMID- 15540242 TI - Agenesis and microdontia of permanent teeth as late adverse effects after stem cell transplantation in young children. AB - BACKGROUND: The objective of the current study was to examine the occurrence of tooth agenesis and microdontia in pediatric stem cell transplantation (SCT) recipients. METHODS: The impact of total body irradiation (TBI) and age at SCT on agenesis and microdontia of permanent teeth was examined in 55 patients from panoramic radiographs. Assessment A1 (for tooth agenesis and microdontia) excluded the third molars, and assessment A2 (for tooth agenesis) included the third molars. Patients were grouped according to TBI status (the TBI group vs. the non-TBI group) and age at SCT (patients age < or = 3.0 years [Group Y], patients ages 3.1-5.0 years [Group M], and patients age > or = 5.1 years [Group O]). RESULTS: From 1 to 12 teeth were missing in 77%, 40%, and 0% of patients (assessment A1) in Groups Y, M, and O, respectively (Group Y vs. Group M, P=0.055; Group Y vs. Group O, P < 0.001; and Group M vs. Group O, P=0.002), increasing to 83%, 78%, and 43%, respectively, when the third molars were included (assessment A2; P values were not significant). Correspondingly, 75%, 60%, and 13%, respectively, of patients had 1-12 microdontic teeth (assessment A1: Group Y vs. Group M, P=0.306; Group Y vs. Group O, P <0.001; and Group M vs. Group O, P=0.003). Recipient age at the time of SCT was found to have a negative correlation with the number of missing teeth (P=0.001) and microdontic teeth (P=0.005). TBI appeared to have little effect on the prevalence of tooth agenesis (assessment A1: TBI group, 32%; non-TBI group, 29%; assessment A2: TBI group, 72%; non-TBI group, 46%; P values were not significant) or on the prevalence of microdontia (assessment A1: TBI, 41%; non-TBI, 50%; P value was not significant). A tendency toward an increased number of affected teeth was noticed in the group of patients who received TBI. CONCLUSIONS: Depending on their age at SCT, 50-100% of pediatric SCT recipients will later present with agenesis and/or microdontia of permanent teeth that may jeopardize occlusal development. Young age (< or = 5.0 years) at SCT was found to be a stronger risk factor than TBI, although TBI caused additive impairment. PMID- 15540243 TI - Granulocyte--colony-stimulating factor (Filgrastim) may overcome imatinib-induced neutropenia in patients with chronic-phase myelogenous leukemia. PMID- 15540244 TI - Synthesis and structure of the highly chlorinated [60]fullerene C60Cl30 with a drum-shaped carbon cage. PMID- 15540245 TI - A metal-free transfer hydrogenation: organocatalytic conjugate reduction of alpha,beta-unsaturated aldehydes. PMID- 15540246 TI - UV-crosslinking of E1 small nucleolar RNA to proteins in frog oocytes. AB - E1/U17 small nucleolar RNA (snoRNA) is a box H/ACA snoRNA. To detect protein bands that UV-crosslink to E1 RNA primarily at uridines, frog oocytes were injected with [alpha-32P]UTP-labeled E1 RNA and incubated, isolated nuclei were UV irradiated, and nuclear contents were digested with RNase A. Wild-type E1 RNA specifically UV-crosslinked to several protein bands. To identify E1 RNA sites involved in these interactions, we tested 21 E1 RNA mutants, each consisting of substitutions in a conserved sequence or structure. UV-crosslinking of different protein bands to E1 RNA depended on one of the following sets of conserved E1 RNA segments: two 5' end RNA sites; five 5' half RNA sites; two 3' half RNA sites; or 14 sites located throughout E1 RNA. Of these conserved E1 RNA sites, UV crosslinking apparently depended on sequences at 11 sites, and structures at 2 sites. Gel electrophoresis with and without RNA competition detected protein bands that are not common to all of the box H/ACA snoRNAs. PMID- 15540247 TI - Prostate carcinoma incidence in relation to prediagnostic circulating levels of insulin-like growth factor I, insulin-like growth factor binding protein 3, and insulin. AB - BACKGROUND: There have been several epidemiologic studies investigating the association between circulating levels of insulin-like growth factor I (IGF-I), insulin-like growth factor binding protein 3 (IGFBP-3), and insulin in relation to the risk of prostate carcinoma, with conflicting results. To examine this issue further, the authors conducted a nested case-control study within the Cardiovascular Health Study cohort. METHODS: In men who were diagnosed with prostate carcinoma (cases) between 1990 and 1999 (n=174), the levels of IGF-I, IGFBP-3, and insulin were measured on blood samples that were obtained 1-9 years prior to diagnosis (mean, 3.4 years). Similar measurements were made on 174 male participants without prostate carcinoma (controls) who were matched to cases based on the year blood was drawn, survival until the date of diagnosis, race, and age. RESULTS: Relative to the men with IGF-I levels in the first (lowest) quartile of the distribution, the risk of prostate carcinoma for men in the second, third, and fourth (upper) quartiles were 0.77 (95% confidence interval [95% CI], 0.43-1.38), 0.73 (95% CI, 0.41-1.30), and 0.67 (95% CI, 0.37-1.25), respectively. The results were influenced little by adjustment for levels of IGFBP-3 or, instead, by evaluating the molar IGF-I/IGFBP-3 ratio. An analysis that was restricted to men who had plasma prostate-specific antigen levels <4 ng/mL at the time of the blood draw yielded similar results. The corresponding relative risks for IGFBP-3 were 0.91 (95% CI, 0.49-1.68), 0.47 (95% CI, 0.25 0.94), and 0.65 (95% CI, 0.35-1.20), respectively. The distribution of serum insulin levels in cases and controls were nearly identical. CONCLUSIONS: The IGF I level was not associated positively with the risk of prostate carcinoma; however, an increase in the IGFBP-3 level was associated with a modest decrease in risk. PMID- 15540248 TI - Differential expression of protein kinase C alpha and delta in testes of mouse at various stages of development. AB - Protein kinase C (PKC) describes a family of serine/threonine protein kinases, and multiple isoforms are expressed in various mammalian tissues. In the present study, we examined the expression of PKC-alpha and PKC-delta at protein and mRNA level in mouse testis by Western blotting and RT-PCR. We also examined the expression of both PKC isoenzymes in the developing mouse testis. In testes of mouse at various developmental stages, both the protein and the mRNA of PKC-alpha were uniformly distributed; but PKC-delta expression occurred in the testes of 3 week-old mice, perhaps even at a relatively late stage in spermatid development. The results suggest that each isoenzyme may have different functional roles in processing and modulating physiological cellular responses of spermatogenesis. PMID- 15540249 TI - Novel biodegradable copolyesters containing blocks of poly(3-hydroxyoctanoate) and poly(epsilon-caprolactone): synthesis and characterization. AB - Novel biodegradable polyester block copolymers have been synthesized by using well-defined poly(3-hydroxyoctanoate) (PHO) oligomers having a hydroxyl end group and an ester end group with M(n) values of 800, 2,500, 5,300, 8,000, or 20,000 as an elastomeric soft segment and poly(epsilon-caprolactone) as a more crystalline segment. These PHO oligomers prepared by methanolysis were subjected to block copolymerization with epsilon-caprolactone. The chemical structure of the copolymers was confirmed by (1)H NMR and (13)C NMR spectroscopy. All the copolyesters are semi-crystalline and two T(g) were observed by differential scanning calorimetry when the molecular weight of the PHO block is about 20,000. PMID- 15540250 TI - Can we change the way we screen?: the ThinPrep Imaging System. PMID- 15540251 TI - Quality assessment in the age of machine-aided cervical cytology screening. PMID- 15540252 TI - Macroscopic spinning chirality memorized in spin-coated films of spatially designed dendritic zinc porphyrin J-aggregates. PMID- 15540254 TI - Effects of simvastatin treatment on oxidant/antioxidant state and ultrastructure of streptozotocin-diabetic rat lung. AB - In the present study, we investigated the effects of simvastatin, a 3-hydroxy-3 methyl-glutaryl coenzyme A reductase inhibitor, on lipid metabolism, lipid peroxidation, antioxidant enzyme activities and ultrastructure of diabetic rat lung. Diabetes was induced by a single injection of streptozotocin (45 mg kg(-1), i.p.). After 8 weeks induction of diabetes, some control and diabetic rats were treated with simvastatin (10 mg kg(-1) rat day(-1); orally) for 4 weeks. Diabetes resulted in significantly high levels of blood glucose and plasma lipids. Malondialdehyde levels were unchanged after 12-week-old diabetic rats, whereas catalase activity significantly decreased in the lung. Glutathione peroxidase activity and nitric oxide level were significantly elevated in the diabetic lung. Histological analysis of the diabetic lung revealed some deterioration in the structure. Simvastatin treatment reduced plasma lipid levels and partially decreased the severity of hyperglycaemia. Catalase, glutathione peroxidase activities and nitric oxide levels were partially restored and accompanied by improved structure in diabetic lung by the simvastatin treatment. These results suggest that structural disturbances and alteration of antioxidative enzyme activities occurred in diabetic lung. Simvastatin treatment may provide some benefits in the maintenance of antioxidant status and structural organization of diabetes-induced injury of lung. PMID- 15540255 TI - Effects on rat testes of the thiosemicarbazone derivative Schiff base (4-(1 phenylmethylcyclobutane-3-yl)-2-(2-hydroxybenzylidenehydrazino)thiazole) and its cadmium(II) complex. AB - The aim of this study was to investigate structural and biochemical changes in testes of rats treated with the thiosemicarbazone derivative thiazole ring Schiff base, (4-(1-phenyl-methylcyclobutane-3-yl)-2-(2-hydroxybenzylidene-hydrazino) thiazole (L), and its Cd(II) complex (CdL(2)). The animals were divided into three groups. Group I was designated as control. The rats in groups II and III were injected subcutaneously with L or CdL(2) respectively at 150-mg kg(-1) doses at 3-day intervals for 15 days. At the end of the study, blood samples were collected for biochemical analysis, and testes were removed for histological examinations. Serum levels of vitamin A, E and MDA of the L-injected group were similar to the control group. While CdL(2) treatment decreased serum vitamin A and E levels, it increased the MDA level compared to other groups. Histologically, the testes structures of L-treated animals were similar to the control. Spermatogenic cells in seminiferous tubules of CdL(2)-treated animals displayed necrosis. Nuclei of spermatogonia and primary spermatocytes were pyknotic and heterochromatic. Homogenous pink particles were present in place of the spermatids. The interstitial areas were oedematous and intertubular vessels were plugged. In conclusion, the present results indicate that L does not cause biochemical and morphological alterations, but its Cd(II) complex has degenerative effects in normal rat testes. PMID- 15540256 TI - Titanium(IV) and zirconium(IV) sulfato complexes containing the Klaui tripodal ligand: molecular models of sulfated metal oxide surfaces. AB - Treatment of titanyl sulfate in about 60 mM sulfuric acid with NaL(OEt) (L(OEt) ( )=[(eta(5)-C(5)H(5))Co{P(O)(OEt)(2)}(3)](-)) afforded the mu-sulfato complex [(L(OEt)Ti)(2)(mu-O)(2)(mu-SO(4))] (2). In more concentrated sulfuric acid (>1 M), the same reaction yielded the di-mu-sulfato complex [(L(OEt)Ti)(2)(mu-O)(mu SO(4))(2)] (3). Reaction of 2 with HOTf (OTf=triflate, CF(3)SO(3)) gave the tris(triflato) complex [L(OEt)Ti(OTf)(3)] (4), whereas treatment of 2 with Ag(OTf) in CH(2)Cl(2) afforded the sulfato-capped trinuclear complex [{(L(OEt))(3)Ti(3)(mu-O)(3)}(mu(3)-SO(4)){Ag(OTf)}][OTf] (5), in which the Ag(OTf) moiety binds to a mu-oxo group in the Ti(3)(mu-O)(3) core. Reaction of 2 in H(2)O with Ba(NO(3))(2) afforded the tetranuclear complex (L(OEt))(4)Ti(4)(mu O)(6) (6). Treatment of 2 with [{Rh(cod)Cl}(2)] (cod=1,5-cyclooctadiene), [Re(CO)(5)Cl], and [Ru(tBu(2)bpy)(PPh(3))(2)Cl(2)] (tBu(2)bpy=4,4'-di-tert-butyl 2,2'-dipyridyl) in the presence of Ag(OTf) afforded the heterometallic complexes [(L(OEt))(2)Ti(2)(O)(2)(SO(4)){Rh(cod)}(2)][OTf](2) (7), [(L(OEt))(2)Ti(O)(2)(SO(4)){Re(CO)(3)}][OTf] (8), and [{(L(OEt))(2)Ti(2)(mu O)}(mu(3)-SO(4))(mu-O)(2){Ru(PPh(3))(tBu(2)bpy)}][OTf](2) (9), respectively. Complex 9 is paramagnetic with a measured magnetic moment of about 2.4 mu(B). Treatment of zirconyl nitrate with NaL(OEt) in 3.5 M sulfuric acid afforded [(L(OEt))(2)Zr(NO(3))][L(OEt)Zr(SO(4))(NO(3))] (10). Reaction of ZrCl(4) in 1.8 M sulfuric acid with NaL(OEt) in the presence Na(2)SO(4) gave the mu-sulfato bridged complex [L(OEt)Zr(SO(4))(H(2)O)](2)(mu-SO(4)) (11). Treatment of 11 with triflic acid afforded [(L(OEt))(2)Zr][OTf](2) (12), whereas reaction of 11 with Ag(OTf) afforded a mixture of 12 and trinuclear [{L(OEt)Zr(SO(4))(H(2)O)}(3)(mu(3)-SO(4))][OTf] (13). The Zr(IV) triflato complex [L(OEt)Zr(OTf)(3)] (14) was prepared by reaction of L(OEt)ZrF(3) with Me(3)SiOTf. Complexes 4 and 14 can catalyze the Diels-Alder reaction of 1,3-cyclohexadiene with acrolein in good selectivity. Complexes 2-5, 9-11, and 13 have been characterized by X-ray crystallography. PMID- 15540257 TI - Synthesis, characterization, and properties of subporphyrazines: a new class of nonplanar, aromatic macrocycles with absorption in the green region. AB - Novel subphthalocyanine analogues that display strong absorption in the green region have been synthesized by using a boron template cyclotrimerization of maleonitrile derivatives. The spectroscopic properties of these macrocycles indicate that, like subphthalocyanines, they have 14 pi electrons and are aromatic compounds with a conical shape. The removal of the three fused benzene rings from the subphthalocyanine skeleton produces a 75-80 nm blue shift of the Q band and a slight lowering of the absorption coefficients for this band. In addition, the reduction of the pi system from 18 to 14 electrons that accompanies progression from porphyrazines to subporphyrazines causes a hypsochromic shift of the Q-band of around 100 nm. Subporphyrazines that are peripherally functionalized with six thioether chains, and in which the sulfur atoms are attached directly to the pyrrole moieties, exhibit optical features that may be explained in terms of the extension of pi conjugation over the six thiolene groups, as well as strong pi donation from the sulfur lone pairs to the macrocycle. These two effects are quantitatively and qualitatively very similar to those observed for porphyrazines that possess the same type of substitution. In addition, the mesomorphic behavior at low temperatures of a macrocycle that is substituted with six thiododecyl chains was demonstrated by using differential scanning calorimetry and optical polarising microscopy. PMID- 15540258 TI - Using diffusion NMR to characterize guanosine self-association: insights into structure and mechanism. AB - This paper presents results from a series of pulsed field gradient (PFG) NMR studies on lipophilic guanosine nucleosides that undergo cation-templated assembly in organic solvents. The use of PFG-NMR to measure diffusion coefficients for the different aggregates allowed us to observe the influences of cation, solvent and anion on the self-assembly process. Three case studies are presented. In the first study, diffusion NMR confirmed formation of a hexadecameric G-quadruplex [G 1](16)4 K(+)4 pic(-) in CD(3)CN. Furthermore, hexadecamer formation from 5'-TBDMS-2',3'-isopropylidene G 1 and K(+) picrate was shown to be a cooperative process in CD(3)CN. In the second study, diffusion NMR studies on 5'-(3,5-bis(methoxy)benzoyl)-2',3'-isopropylidene G 4 showed that hierarchical self-association of G(8)-octamers is controlled by the K(+) cation. Evidence for formation of both discrete G(8)-octamers and G(16)-hexadecamers in CD(2)Cl(2) was obtained. The position of this octamer-hexadecamer equilibrium was shown to depend on the K(+) concentration. In the third case, diffusion NMR was used to determine the size of a guanosine self-assembly where NMR signal integration was ambiguous. Thus, both diffusion NMR and ESI-MS show that 5'-O acetyl-2',3'-O-isopropylidene G 7 and Na(+) picrate form a doubly charged octamer [G 7](8)2 Na(+)2 pic(-) 9 in CD(2)Cl(2). The anion's role in stabilizing this particular complex is discussed. In all three cases the information gained from the diffusion NMR technique enabled us to better understand the self-assembly processes, especially regarding the roles of cation, anion and solvent. PMID- 15540259 TI - Novel carbohydrate-appended metal complexes for potential use in molecular imaging. AB - Seven discrete sugar-pendant diamines were complexed to the {M(CO)(3)}(+) ((99m)Tc/Re) core: 1,3-diamino-2-propyl beta-D-glucopyranoside (L(1)), 1,3 diamino-2-propyl beta-D-xylopyranoside (L(2)), 1,3-diamino-2-propyl alpha-D mannopyranoside (L(3)), 1,3-diamino-2-propyl alpha-D-galactopyranoside (L(4)), 1,3-diamino-2-propyl beta-D-galactopyranoside (L(5)), 1,3-diamino-2-propyl beta (alpha-D-glucopyranosyl-(1,4)-D-glucopyranoside) (L(6)), and bis(aminomethyl)bis[(beta-D-glucopyranosyloxy)methyl]methane (L(7)). The Re complexes [Re(L(1)-L(7))(Br)(CO)(3)] were characterized by (1)H and (13)C 1D/2D NMR spectroscopy which confirmed the pendant nature of the carbohydrate moieties in solution. Additional characterization was provided by IR spectroscopy, elemental analysis, and mass spectrometry. Two analogues, [Re(L(2))(CO)(3)Br] and [Re(L(3))(CO)(3)Br], were characterized in the solid state by X-ray crystallography and represent the first reported structures of Re organometallic carbohydrate compounds. Conductivity measurements in H(2)O established that the complexes exist as [Re(L(1)-L(7))(H(2)O)(CO)(3)]Br in aqueous conditions. Radiolabelling of L(1)-L(7) with [(99m)Tc(H(2)O)(3)(CO)(3)](+) afforded in high yield compounds of identical character to the Re analogues. The radiolabelled compounds were determined to exhibit high in vitro stability towards ligand exchange in the presence of an excess of either cysteine or histidine over a 24 h period. PMID- 15540260 TI - Aggregation of [Cu(II)4] building blocks into [Cu(II)8] clusters or a [Cu(II)4]infinity chain through subtle chemical control. AB - Coordination complexes of the ligand H3L [1,3-bis(3-oxo-3-phenylpropionyl)-2 hydroxy-5-methylbenzene] with Cu(II) are reported. Clusters showing various nuclearities or modes of supramolecular organization have been prepared by slightly changing the reaction conditions and have been crystallographically characterized. The reaction of H3L with one equivalent of Cu(OAc)2 in DMF yields the dinuclear complex [Cu2(HL)2(dmf)2] (1). Reaction in MeOH of H3L with an increased amount of metal, in the form of Cu(NO3)2, and excess strong base (nBu4NOH) affords the cluster [Cu8(L)2(OMe)8(NO3)2] (2). Complex 2 is a dimer of two linear [Cu4] arrays bridged by methoxide ligands, where the polynucleating ligand is fully deprotonated. The [Cu4]2 clusters are linked to each other by NO3 bridges to form one-dimensional coordination polymers. The link between [Cu8] units and their relative spatial positioning can be modified by changing the anion of the Cu(II) salt, as demonstrated by the synthesis of the cluster polymers [Cu8(L)2(OMe)8Cl2] (3) and [Cu8(L)(OMe)7.86Br2.14] (4), where only NO3- has been replaced by Cl- or Br-, respectively. Similarly, when ClO4- is used, compound [Cu8(L)2(OMe)8(ClO4)2(MeOH)4] (5) can be isolated. It contains independent [Cu8] units. A slight change in the stoichiometry of the reaction leading to 2 affords the related complex catena-[Cu4(L)(OMe)3(NO3)2(H2O)0.36] (6). This polymer contains essentially the same [Cu4] moiety as 2, albeit organized in a completely different arrangement. Each [Cu4] unit in 6 is linked by OMe- ligands to two such equivalent groups to form an infinite chain. Magnetic susceptibility measurements reveal weak antiferromagnetic exchange between Cu(II) centers in 1 (J = -0.73 cm(-1)) and strong antiferromagnetic coupling within [Cu4] chains in 2, 5, and 6 (most negative J values of -113.8 and -177.3 cm(-1) for 2 and 6, respectively). PMID- 15540261 TI - Discrepancy between the spin distribution and the magnetic ground state for a triaminoxyl substituted triphenylphosphine oxide derivative. AB - The magnetic interaction and spin transfer via phosphorus have been investigated for the tri-tert-butylaminoxyl para-substituted triphenylphosphine oxide. For this radical unit, the conjugation existing between the pi* orbital of the NO group and the phenyl pi orbitals leads to an efficient delocalization of the spin from the radical to the neighboring aromatic ring. This has been confirmed by using fluid solution high-resolution EPR and solid state MAS NMR spectroscopy. The spin densities located on the atoms of the molecule could be probed since (1)H, (13)C, (14)N, and (31)P are nuclei active in NMR and EPR, and lead to a precise spin distribution map for the triradical. The experimental investigations were completed by a DFT computational study. These techniques established in particular that spin density is located at the phosphorus (rho=-15x10(-3) au), that its sign is in line with the sign alternation principle and that its magnitude is in the order of that found on the aromatic C atoms of the molecule. Surprisingly, whereas the spin distribution scheme supports ferromagnetic interactions among the radical units, the magnetic behavior found for this molecule revealed a low-spin ground state characterized by an intramolecular exchange parameter of J=-7.55 cm(-1) as revealed by solid state susceptibility studies and low temperature EPR. The X-ray crystal structures solved at 293 and 30 K show the occurrence of a crystallographic transition resulting in an ordering of the molecular units at low temperature. PMID- 15540262 TI - Electronic structure and chain-length effects in diplatinum polyynediyl complexes trans,trans-[(X)(R3P)2Pt(C triple bond C)(n)Pt(PR3)2(X)]: a computational investigation. AB - Structure and bonding in the title complexes are studied using model compounds trans,trans-[(C6H5)(H3P)2Pt(C triple bond C)(n)Pt(PH3)2(C6H5)] (PtCxPt; x = 2n = 4-26) at the B3LYP/LACVP* level of density functional theory. Conformations in which the platinum square planes are parallel are very slightly more stable than those in which they are perpendicular (DeltaE = 0.12 kcal mol(-1) for PtC8Pt). As the carbon-chain length increases, progressively longer C triple bond C triple bonds and shorter triple bond C-C triple bond single bonds are found. Whereas the triple bonds in HCxH become longer (and the single bonds shorter) as the interior of the chain is approached, the PtC triple bond C triple bonds in PtCxPt are longer than the neighboring triple bond. Also, the Pt-C bonds are shorter at longer chain lengths, but not the H-C bonds. Accordingly, natural bond orbital charge distributions show that the platinum atoms become more positively charged, and the carbon chain more negatively charged, as the chain is lengthened. Furthermore, the negative charge is localized at the two terminal C triple bond C atoms, elongating this triple bond. Charge decomposition analyses show no significant d-pi* backbonding. The HOMOs of PtCxPt can be viewed as antibonding combinations of the highest occupied pi orbital of the sp-carbon chain and filled in-plane platinum d orbitals. The platinum character is roughly proportional to the Pt/Cx/Pt composition (e.g., x = 4, 31 %; x = 20, 6 %). The HOMO and LUMO energies monotonically decrease with chain length, the latter somewhat more rapidly so that the HOMO-LUMO gap also decreases. In contrast, the HOMO energies of HCxH increase with chain length; the origin of this dichotomy is analyzed. The electronic spectra of PtC4Pt to PtC10Pt are simulated. These consist of two pi pi* bands that redshift with increasing chain length and are closely paralleled by real systems. A finite HOMO-LUMO gap is predicted for PtCinfinityPt. The structures of PtCxPt are not strictly linear (average bond angles 179.7 degrees 178.8 degrees ), and the carbon chains give low-frequency fundamental vibrations (x = 4, 146 cm(-1); x = 26, 4 cm(-1)). When the bond angles in PtC12Pt are constrained to 174 degrees in a bow conformation, similar to a crystal structure, the energy increase is only 2 kcal mol(-1). The above conclusions should extrapolate to (C triple bond C)(n) systems with other metal endgroups. PMID- 15540263 TI - Chiral molecular glass: synthesis and characterization of enantiomerically pure thiophene-based [7]helicene. AB - Synthesis of thiophene-based [7]helicenes, which are functionalized for both design of organic chiral glasses with strong chiroptical properties and for further homologation to higher [n]helicenes, is reported. The key synthetic transformations are kinetic resolution of the intermediate diketone and the annelation step forming the center benzene ring by means of an intramolecular McMurry reaction. Based upon X-ray crystallographic determinations of the absolute configurations for (+)-enantiomers of the diketone and the [7]helicene, stereochemical correlation between the (R) axial chirality of the diketone and the (M) helical chirality of the [7]helicene is established. One such enantiopure trimethylsilyl-substituted [7]helicene possesses enchanced chiroptical properties and forms a chiral molecular glass. PMID- 15540264 TI - An electronic perspective on the reduction of an n=n double bond at a conserved dimolybdenum core. AB - Density functional theory has been used to assess the role of the bimetallic core in supporting reductive cleavage of the N=N double bond in [Cp2Mo2(mu-SMe)3(mu eta1:eta1-HN=NPh)]+. The HOMO of the complex, the Mo-Mo delta orbital, plays a key role as a source of high-energy electrons, available for transfer into the vacant orbitals of the N=N unit. As a result, the metal centres cycle between the Mo(III) and Mo(IV) oxidation states. The symmetry of the Mo-Mo delta "buffer" orbital has a profound influence on the reaction pathway, because significant overlap with the redox-active orbital on the N=N unit (pi* or sigma*) is required for efficient electron transfer. The orthogonality of the Mo-Mo delta and N-N sigma* orbitals in the eta1:eta1 coordination mode ensures that electron transfer into the N-N sigma bond is effectively blocked, and a rate-limiting eta1:eta1- >eta1 rearrangement is a necessary precursor to cleavage of the bond. PMID- 15540265 TI - First detection of a selenenyl fluoride ArSe-F by NMR spectroscopy: the nature of Ar2Se2/XeF2 and ArSe-SiMe3/XeF2 reagents. AB - Arylselenenyl fluorides ArSeF are obtained from diselenides Ar2Se2 or arylselenotrimethylsilanes ArSe-SiMe3, and XeF2. They are detected by low temperature 19F and 77Se NMR spectroscopy. Substitution in the ortho position of the aromatic ring to provide electronic or steric protection is a requirement for their formation. ArSe--F compounds decompose according to 3 ArSe-F-->[ArSe SeF2Ar]+ArSe-F-->ArSeF3+Ar2Se2. Reaction energies for this disproportionation as well as that of the sulfur and tellurium homologues have been calculated with MP2, CCSD(T,) and B3 LYP methods. They were found to be increasingly exothermic in the sequence SE+Arg P, K=1.0; E.Arg-->E+Arg, K=0.45; E.MgATP-->E+MgATP, K=0.17; E.MgADP- >E+MgADP, K=0.12; and the ternary complex: Arg P.E.MgADP-->E.MgADP+Arg P, K=0.94; Arg.E.MgATP-->E.MgATP+Arg, K=0.49; MgATP.Enz.Arg-->E.Arg+MgATP, K=0.14; MgADP.E.Arg P-->E.Arg P+MgADP, K=0.09. For a particular substrate, the ratio of the dissociation constants for the binary to ternary complex is close to one, indicating little, if any, cooperativity in substrate binding for the rapid equilibrium, random addition mechanism. The time course of the arginine kinase reaction exhibits a pronounced curvature, which, as described for enzyme from other sources, is attributed to formation of an inhibitory catalytic dead-end complex, MgADP.E.Arg. The curvature is accentuated by the addition of monovalent anions, including borate, thiocyanate, and, most notably, nitrite and nitrate. This effect is attributed to stabilization of the dead-end complex through formation of a transition state analog. However, the substantial decrease in initial velocity (92%) caused by nitrate is due to an additional inhibitory effect, further characterized as non-competitive inhibition (Ki=8.0 mM) with the substrate L-arginine. On the other hand, borate inhibition of the initial velocity is only 30% with significant subsequent curvature, suggesting that this anion functions as an inhibitor mainly by formation of a transition state analog. However, some component of the borate inhibition appears to be mediated by an apparent partial competitive inhibition with L-arginine. D-arginine is not a substrate for arginine kinase from the cockroach, but is an effective competitive inhibitor with a Ki=0.31 mM. L-Canavanine is a weak substrate for arginine kinase (Km=6.7 mM) with a Vmax for the pure enzyme that is approximately one-third that of L-arginine. However, initial velocity experiments of substrate mixtures suggest that competition between L-canavanine and L-arginine may not be a simple summation effect and may involve a structural modification. Sensitivity of arginine kinase activity to D-arginine as well as nitrate and borate anions, coupled with the fact that L-arginine is an essential amino acid for the cockroach, suggest that arginine kinase could be a useful chemotherapeutic target for the control of cockroach proliferation. PMID- 15540276 TI - Endopeptidase activity of larval Lacanobia oleracea corpus allatum: metabolism of Manduca sexta allatostatin and allatotropin. AB - The degradation of synthetic Manduca sexta allatostatin (Manse-AS) and allatotropin (Manse-AT) by enzymes associated with the corpus allatum (CA) of larvae of the tomato moth, Lacanobia oleracea, was investigated using reversed phase high performance liquid chromatography and matrix-assisted laser desorption ionisation-time of flight mass spectrometry. Manduca sexta allatostatin was metabolised by CA extract to Manse-AS5-15, Manse-AS6-15, and Manse-AS7-15, which indicates enzymic cleavage at the C-terminal side of arginine residues R3 and R5 and the N-terminal side of R5, suggesting this is due to a trypsin-like enzyme. In support of this, the same degradation products were identified after Manse-AS was incubated with trypsin, and CA enzymic activity could be inhibited up to 79% by aprotinin. Degradation of Manse-AT by CA extract was also trypsin-like, cleaving at the C-terminal side of the basic residues K3 and R11 to produce Manse AT4-13 and Manse-AT1-11. Metabolism by trypsin produced the same deletion peptides, but the major product due to this enzyme was Manse-AT4-11. Hydrolysis of Manse-AT by CA could only be partially inhibited by high doses of aprotinin (36%), and the CA extract also cleaved Manse-AT between M8 and T9 to produce Manse-AT1-8. A trypsin-like peptidase appears to be the major enzyme present in the CA of larval L. oleracea that acts to metabolise Manse-AS and Manse-AT. In addition, an unidentified enzyme that cleaves between M and T residues degraded Manse-AT. PMID- 15540277 TI - Ornithine decarboxylase activity during the development of Anastrepha fraterculus (Diptera, Tephritidae). AB - Ornithine decarboxylase (ODC) (EC 4.1.1.17) is very important for polyamine biosynthesis, which is required for main biological events. In the present study, ODC activity was measured in samples of Anastrepha fraterculus's egg, larva, pupa body and abdomen, adult body, ovaries, and fat body of young females, and in ovaries of mature flies. The kinetic parameters (Km app and Vmax) for ODC activity were determined for pupa, larva, and young ovary. ODC activity showed fluctuations during A. fraterculus's life development. In its earlier stages, prior to emergence, the egg has high ODC-specific activity probably due to embryogenesis, which is characterized by a high rate of cell division. This enzyme activity is also significantly high in the ovary and fat body of young females possibly related to the increased oogenesis and vitellogenesis. The kinetic parameters (Km app and Vmax) had great variation. Our results using GTP showed that the great variation in kinetic parameters can be accounted for by post-translational modifications. PMID- 15540278 TI - Toxicity of two type II ribosome-inactivating proteins (cinnamomin and ricin) to domestic silkworm larvae. AB - Cinnamomin and ricin are two type II ribosome-inactivating proteins. They exhibited a different toxicity to domestic silkworm (Bombyx mori) larvae by oral feeding bioassay. The LC50 of ricin to the silkworm larvae at third instar was much lower than that of cinnamomin. When the isolated 80S ribosome from domestic silkworm pupae was treated separately with the reduced cinnamomin or the reduced ricin, a specific RNA fragment (R-fragment) was produced as characterized by 8 M urea-denatured polyacrylamide gel (3.5%) electrophoresis. The purified A-chains of both cinnamomin and ricin showed a slightly different RNA N-glycosidase activity to the domestic silkworm pupal ribosome. It was proposed that the difference of their toxicity to domestic silkworm larvae was not related to their A-chains but to the properties of their B-chains. It was also found that the vomit obtained from the midgut of domestic silkworm larvae could hydrolyze these two proteins apparently to a similar extent. PMID- 15540280 TI - Problems along the new "silk road". PMID- 15540279 TI - Biologically motivated computational modeling: contribution to risk assessment. PMID- 15540282 TI - Living related kidney and liver transplantation. 27 June 2003, Heidelberg, Germany. Proceedings. PMID- 15540281 TI - Apolipoprotein A-I infiltration in rheumatoid arthritis synovial tissue: a control mechanism of cytokine production? AB - The production of tumor necrosis factor alpha (TNF-alpha) and interleukin-1beta (IL-1beta) by monocytes is strongly induced by direct contact with stimulated T lymphocytes, and this mechanism may be critical in the pathogenesis of rheumatoid arthritis (RA). Apolipoprotein A-I (apoA-I) blocks contact-mediated activation of monocytes, causing inhibition of TNF-alpha and IL-1beta production. This study examined the hypothesis that apoA-I may have a regulatory role at sites of macrophage activation by T lymphocytes in inflamed RA synovial tissue. Synovial tissue samples were obtained after arthroscopy from patients with early untreated RA or treated RA and from normal subjects. As determined by immunohistochemistry, apoA-I was consistently present in inflamed synovial tissue that contained infiltrating T cells and macrophages, but it was absent from noninflamed tissue samples obtained from treated patients and from normal subjects. ApoA-I staining was abundant in the perivascular areas and extended in a halo-like pattern to the surrounding cellular infiltrate. C-reactive protein and serum amyloid A were not detected in the same perivascular areas of inflamed tissues. The abundant presence of apoA-I in the perivascular cellular infiltrates of inflamed RA synovial tissue extends the observations in vitro that showed that apoA-I can modify contact-mediated macrophage production of TNF-alpha and IL-1beta. ApoA-I was not present in synovium from patients in apparent remission, suggesting that it has a specific role during phases of disease activity. These findings support the suggestion that the biologic properties of apoA-I, about which knowledge is newly emerging, include anti-inflammatory activities and therefore have important implications for the treatment of chronic inflammatory diseases. PMID- 15540283 TI - Minnesota Colorectal Cancer Initiative: successful development and implementation of a community-based colorectal cancer registry. AB - PURPOSE: The aim of the Minnesota Colorectal Cancer Initiative is to implement risk-specific interventions to decrease colorectal cancer morbidity and mortality by 1) assisting clinicians to identify and educate individuals and families at high and increased risk for colorectal cancer; 2) providing professional and community education; 3) maintaining a database to evaluate the effectiveness of preventive intervention strategies; and 4) facilitating colorectal cancer research. METHODS: Two physician groups and the University Cancer Center founded the Minnesota Colorectal Cancer Initiative as a not-for-profit organization. Health care organizations, pharmaceutical companies, a consulting firm, and other practice groups provide continuing financial and other support. A database registry, risk-assessment survey, and consent document were developed and then were approved by an institutional review board. A trial enrollment was conducted. Minnesota Colorectal Cancer Initiative services are available to the public. Participants are actively recruited through member organizations. Minnesota Colorectal Cancer Initiative assesses hereditary risk and will document family history in the medical record on request. A personally targeted reply letter reviews risk factors and recommends specific screening and surveillance strategies for participants and their family members, and when appropriate, provides information regarding genetic counseling and testing services. Minnesota Colorectal Cancer Initiative services are free to participants. RESULTS: Since 1999, Minnesota Colorectal Cancer Initiative has sent individually tailored reply letters providing risk-specific information about colorectal cancer to 717 participants and more than 3200 of their first-degree and second-degree relatives. More than 200 families, previously unidentified as having histories suggestive of hereditary colorectal cancer (attenuated familial polyposis and hereditary nonpolyposis colorectal cancer), have been identified; genetic services were explained and recommended. A formal program evaluation confirmed that Minnesota Colorectal Cancer Initiative provides useful information and materials and promotes intrafamilial communication about colon cancer risk and recommendations. CONCLUSIONS: Minnesota Colorectal Cancer Initiative is a model of effective collaboration between academic and community health care providers. A community-based registry is a unique way to identify and provide personal, risk specific information to large numbers of people at increased or high risk for colorectal cancer. PMID- 15540284 TI - Comparison of long-term functional results of colonic J-pouch and straight anastomosis after low anterior resection for rectal cancer: a five-year follow up. AB - PURPOSE: Few reports on the long-term functional outcome of colonic J-pouch reconstruction have been published, and data comparing J-pouch and straight reconstruction are contradictory. This prospective study compares the functional outcome of colonic J-pouch and straight anastomosis five years after low anterior resection for rectal cancer. METHODS: Functional outcome was compared in 46 patients with J-pouch reconstruction (J-group) and 48 patients with straight anastomosis (S-group). Clinical status was evaluated with a 17-item questionnaire inquiring about different aspects of bowel function. Reservoir function was evaluated by manovolumetry. The Fisher's exact test and Wilcoxon's rank-sum test were used to compare categoric and quantitative data, respectively. RESULTS: Among patients with an ultralow anastomosis (< or = 4 cm from the anal verge), the number of bowel movements during the day (> or = 5, 4.3 vs. 29.2 percent; P = 0.028) and at night (> 1/week, 4.3 vs. 33.3 percent; P = 0.013) and urgency (4.3 vs. 33.3 percent; P = 0.013) and soiling (21.7 vs. 50.0 percent; P = 0.043) were less in the J-group than in the S-group. Among patients with a low anastomosis (5 to 8 cm from the verge), patients in the J-group had fewer bowel movements at night (> 1/week, 0 vs. 20.8 percent; P = 0.028) and less urgency (0 vs. 20.8 percent; P = 0.028). Reservoir function was better in the J-group than in the S group in both the ultralow (maximum tolerable volume (mean), 101.7 vs. 76.3 ml; P = 0.004; threshold volume (mean), 46.5 vs. 30.4 ml; P < 0.001; compliance (mean), 4.9 vs. 2.5 ml/cm H2O; P < 0.001) and low-anastomosis (maximum tolerable volume, 120.4 vs. 97.9 ml; P < 0.001; threshold volume, 58.3 vs. 40.8 ml; P < 0.001; compliance, 5.2 vs. 3.1 ml/cm H2O; P < 0.001) groups. CONCLUSIONS: J-pouch reconstruction increased reservoir function and provided better functional outcome than straight anastomosis, even five years after surgery, especially in patients whose anastomosis is less than 4 cm from the anal verge. PMID- 15540285 TI - Transverse coloplasty pouch after total mesorectal excision: functional assessment of evacuation. AB - PURPOSE: Colon pouch reconstruction after total mesorectal excision is functionally superior to straight colorectal/ anal anastomosis. In the long-term, stool evacuation difficulties could jeopardize the functional benefit. The transverse coloplasty pouch presents an alternative to the standard J-pouch. This study was designed to analyze functional outcome and defecography findings after total mesorectal excision and transverse coloplasty pouch reconstruction. METHODS: Thirty consecutive patients with cancer of the middle and lower third of the rectum underwent a total mesorectal excision and were examined in a prospective study. In all patients, reconstruction was performed with a transverse coloplasty pouch. Pouch and anastomosis were checked by Gastrografin enema postoperatively. Patients were examined within eight months by means of defecography, manometry, pouch volumetry, and a standardized continence questionnaire. RESULTS: Total mesorectal excision with transverse coloplasty pouch anastomosis was performed successfully in all patients. Symptomatic anastomotic leakage was observed in 2 of 30 patients and the radiologic leak rate was 4 of 30. All patients evacuated the pouch completely; none needed enemas or suppositories to facilitate defecation. Twenty-five of 27 patients had a maximum of three bowel movements per day, and all patients were continent for solid stools. Patients with abnormal findings on defecography proved more likely to have anal dysfunction. CONCLUSIONS: Transverse coloplasty pouch reconstruction after total mesorectal excision leads to good functional results and is not associated with stool evacuation problems. Urgency and incontinence correlate rather with impaired pelvic floor movement than with pouch size or anal sphincter tonus. PMID- 15540286 TI - Pilot study of the quality initiative in rectal cancer strategy. AB - INTRODUCTION: Total mesorectal excision vs. traditional surgical techniques may lead to improved rates of permanent colostomy, local tumor recurrence, and survival for patients undergoing major rectal cancer operations. We developed the surgeon-directed, multipronged Quality Initiative in Rectal Cancer strategy to encourage surgeons to use total mesorectal excision techniques. METHODS: The Quality Initiative in Rectal Cancer strategy interventions included a workshop, an operative demonstration of total mesorectal excision, and a postoperative questionnaire. The design of the strategy was informed by the industrial theory principles of continuous quality improvement. We assessed the logistics of implementing the strategy and the attitudes of surgeons toward the strategy through a pilot study at three community hospitals in the Central-West region of Ontario. RESULTS: Seventeen of 19 surgeons participated in a workshop, and 12 of 17 workshop participants received at least one operative demonstration of total mesorectal excision. Ten of 11 surgeons who completed a postoperative questionnaire indicated their traditional approach to rectal cancer surgery varied with that of the operative demonstration. The attitudes of surgeons toward the Quality Initiative in Rectal Cancer strategy were positive. For the time periods before and after the pilot study, there was a trend toward a lower rate of permanent colostomy among patients treated by surgeons who participated in both the workshop and an operative demonstration of total mesorectal excision. CONCLUSION: The Quality Initiative in Rectal Cancer strategy may be an effective method of introducing optimal rectal cancer surgery techniques to a large group of practicing surgeons. PMID- 15540287 TI - Colorectal cancer pelvic recurrences: determinants of resectability. AB - PURPOSE: This study was designed to identify preoperative and intraoperative features of locally recurrent colorectal cancer that predict R0 resection in patients scheduled for attempted complete resection followed by intraoperative radiation therapy. METHODS: Review of a prospective data base identified 119 patients brought to the intraoperative radiation therapy suite for planned complete resection of locally recurrent rectal (n = 101) and colon (n = 18) cancer between January 1994 and November 2000. R0 resection was achieved in 61 patients. This group was compared with patients in which an R1 (n = 38), R2 (n = 7), or palliative procedure (n = 13) was performed. Variables evaluated included: tumor location, features of the primary tumor, and preoperative findings on computed tomography, magnetic resonance imaging, and history/physical. Tumor location was established by review of operative/pathologic reports and classified as axial (anastomotic/perineal), anterior (bladder/genitourinary organs), posterior (presacral), or lateral (pelvic sidewall). RESULTS: When recurrence was confined to the axial location only, or axial and anterior locations, R0 resection was achieved significantly more often than when other locations were involved (P < 0.001, P = 0.003, respectively). When a lateral component was present, R0 resection was achieved significantly less often than when there was no lateral component (P = 0.002). For patients with available preoperative computed tomography and/or magnetic resonance imaging results (n = 70), the finding of lateral tumor involvement was associated with R0 resection significantly less often than when lateral disease was not identified (P = 0.004). CONCLUSIONS: Pelvic recurrences confined to the axial location, or axial and anterior locations, are more likely to be completely resectable (R0) than those involving the pelvic sidewall. Efforts to enhance preoperative identification and imaging of these patients are clearly justified. PMID- 15540288 TI - Does rectal wall tumor eradication with preoperative chemoradiation permit a change in the operative strategy? AB - PURPOSE: Preoperative chemoradiation may downstage locally advanced rectal cancer and, in some cases, with no residual tumor. The management of complete response is controversial and recent data suggest that radical surgery may be avoided in selected cases. Transanal excision of the scar may determine the rectal wall response to chemoradiation. This study was designed to assess whether the absence of tumor in the bowel wall corresponds to the absence of tumor in the mesorectum, known as true complete response. METHODS: A retrospective review of the medical records of patients who underwent preoperative chemoradiation for advanced mid (6 11 cm from the anal verge) and low (from the dentate line to 5 cm from the anal verge) rectal cancer (uT2-uT3) followed by radical surgery with total mesorectal excision was undertaken. Patients in whom the pathology specimen showed no residual tumor in the rectal wall (yT0, "y" signifies pathologic staging in postradiation patients) were assessed for tumoral involvement of the mesorectum. RESULTS: A total of 109 patients underwent preoperative, high-dose radiation therapy (94 percent with 5-fluorouracil chemosensitization), followed by radical surgery for advanced rectal cancer. Preoperatively, 47 patients were clinically assessed to have potentially complete response. After radical rectal resection, pathology did not reveal any residual tumor within the rectal wall (yT0) in 17 patients. In two (12 percent) of these patients, the mesorectum was found to be positive for malignancy: one had positive lymph nodes that harbored cancer; one had tumor deposits in the mesorectal tissue. CONCLUSIONS: Compete rectal wall tumor eradication does not necessarily imply complete response, because the mesorectum may harbor tumor cells. Thus, caution should be exercised when considering the avoidance of radical surgery. Reliable imaging methods and clinical predictors for favorable outcome are important to allow less radical approaches in the future. PMID- 15540289 TI - The fate of the ileal pouch in patients developing Crohn's disease. AB - PURPOSE: Recent studies have suggested that a subset of patients with Crohn's colitis may have a favorable outcome after ileal pouch-anal anastomosis and have advocated elective ileal pouch-anal anastomosis in selected patients with Crohn's disease. We have not offered ileal pouch-anal anastomosis to patients with known Crohn's disease, but because of the overlap in clinical presentation of ulcerative colitis and indeterminate colitis, some patients receiving an ileal pouch-anal anastomosis are subsequently found to have Crohn's disease. We review our experience with these patients to identify potential preoperative predictors of ultimate pouch failure. METHODS: Patients with a final diagnosis of Crohn's disease were identified from an ileal pouch-anal anastomosis registry. These patients are followed prospectively. Preoperative and postoperative clinical and pathologic characteristics were evaluated as predictors of outcome. Median (range) values are listed. RESULTS: Thirty-two (18 females) patients (4.1 percent) with a final diagnosis of Crohn's disease were identified from a registry of 790 ileal pouch-anal anastomosis patients (1980-2002). Patients underwent ileal pouch-anal anastomosis in two stages (11 patients) or three stages (21 patients). The preoperative diagnosis was ulcerative colitis in 24 patients and indeterminate colitis in 8 patients. Median follow-up was 153 (range, 13-231) months. The median time from ileal pouch-anal anastomosis to diagnosis of Crohn's disease was 19 (range, 0-188) months. Complications occurred in 93 percent, including perineal abscess/fistula (63 percent), pouchitis (50 percent), and anal stricture (38 percent). Pouch failure (excision or current diversion) occurred in nine patients (29 percent) at a median of 66 (range, 6 187) months. Two of these 9 patients had preoperative anal disease (not significant). Comparing patients with failed pouches (n = 9) to patients with functioning pouches (n = 23), post-ileal pouch-anal anastomosis perineal abscess (67 vs. 26 percent, P = 0.05) and pouch fistula (89 vs. 30 percent, P = 0.01) were more commonly associated with pouch failure. Preoperative clinical, endoscopic, and pathologic features were not predictive of pouch failure or patient outcome. For those with a functional pouch, 50 percent have been or are currently on medication to treat active Crohn's disease. This group had six bowel movements in 24 (range, 3-10) hours, with leakage in 60 percent and pad usage in 45 percent. CONCLUSIONS: Patients who undergo ileal pouch-anal anastomosis and are subsequently found to have Crohn's disease experience significant morbidity. Preoperative characteristics, including the presence of anal disease, were not predictive of subsequent pouch failure. We choose not to recommend the routine application of ileal pouch-anal anastomosis in any subset of patients with known Crohn's disease. PMID- 15540290 TI - Outcome after colectomy for Clostridium difficile colitis. AB - PURPOSE: Clostridium difficile colitis is a relatively common entity, yet large series of patients with fulminant C. difficile colitis are infrequently reported. This study was designed to identify risk factors, clinical characteristics, and outcome of patients who required colectomy for fulminant C. difficile colitis. METHODS: A population-based study on all patients in 159 hospitals of the Department of Veterans Affairs from 1997 to 2001 was performed. Data were compiled from several national computerized Department of Veterans Affairs data sets. Supplementary information including demographic information, discharge summaries, operative reports, and pathology reports were obtained from local medical records. Patient variables were entered into a computerized database and analyzed using the Pearson chi-squared and Fisher's exact tests. Statistical significance was designated as P < 0.05. RESULTS: Sixty-seven patients (mean age, 69 (range, 40-86) years; 99 percent males) were identified. All 67 patients had C. difficile verified in the colectomy specimens. Thirty-six of 67 patients (54 percent) developed C. difficile colitis during a hospitalization for an unrelated illness, and 30 of 36 patients (87 percent) after a surgical procedure. Thirty one of 67 (46 percent) developed C. difficile colitis at home. There was no history of diarrhea in 25 of 67 patients (37 percent). Thirty of 67 patients (45 percent) presented in shock (blood pressure, <90 mmHg). Forty-three of 67 patients (64 percent) presented with an acute surgical abdomen. Mean white blood cell count was 27.2; mean percent bands was 12. Twelve of 67 patients (18 percent) had a negative C difficile colitis stool assay. Abdominal computed tomography correctly diagnosed 45 of 46 patients (98 percent) who were imaged. Twenty-six of 67 patients (39 percent) underwent colonoscopy; all 26 were found to have severe inflammation or pseudomembranes. Fifty-three of 67 patients (80 percent) underwent total colectomy; 14 of 67 underwent segmental colonic resection. Perforation and infarction were found in 59 of 67 patients (58 percent) at surgery. Overall mortality was 48 percent (32/67). Mean hospitalization was 36 (range, 2-297) days. CONCLUSIONS: Patients with fulminant C. difficile colitis often present with an unexplained abdominal illness with a marked leukocytosis that rapidly progresses to shock and peritonitis. Although frequently developed during a hospitalization and often after a surgical procedure, it may develop outside of a hospital setting. Diarrhea may be absent and stool cytology may be negative for C. difficile toxin. Perforation and infarction are frequently found at surgery. In those patients who survive, a prolonged hospitalization is common. Mortality from fulminant C. difficile colitis remains high despite surgical intervention. PMID- 15540291 TI - Subtypes of anal incontinence associated with bowel dysfunction: clinical, physiologic, and psychosocial characterization. AB - BACKGROUND: We hypothesized that functional anal incontinence with no structural explanation comprises distinct pathophysiologic subgroups that could be identified on the basis of the predominant presenting bowel pattern. METHODS: Consecutive patients (n = 80) were prospectively grouped by bowel symptoms as 1) incontinence only, 2) incontinence + constipation, 3) incontinence + diarrhea, and 4) incontinence + alternating bowel symptoms. The Hopkins Bowel Symptom Questionnaire, the Symptom Checklist 90-R, and anorectal manometry were completed. RESULTS: Significant group differences were found between subcategories of incontinent patients on the basis of symptoms. Abdominal pain was more frequent in patients with altered bowel patterns. Patients with alternating symptoms reported the highest prevalence of abdominal pain, rectal pain, and bloating. Basal anal pressures were significantly higher in alternating patients (P = 0.03). Contractile pressures in the distal anal canal were diminished in the incontinent-only and diarrhea groups (P = 0.004). Constipated patients with incontinence exhibited elevated thresholds for the urge to defecate (P = 0.027). Dyssynergia was significantly more frequent in patients with incontinence and constipation or alternating bowel patterns. CONCLUSIONS: Distinct patterns of pelvic floor dysfunction were identified in patient subgroups with anal incontinence, based on the presence or absence of altered bowel patterns. Physiologic assessments suggested different pathophysiologic mechanisms among the subgroups. The evaluation of patients with fecal incontinence should consider altered bowel function. PMID- 15540292 TI - Transanal or vaginal approach to rectocele repair: a prospective, randomized pilot study. AB - PURPOSE: This study was designed to compare outcomes of transanal and vaginal techniques for rectocele repair. METHODS: Thirty females with symptomatic rectocele were enrolled in a prospective, randomized study. Fifteen underwent transanal rectoceleplasty, the other 15 underwent vaginal posterior colporrhaphy. Patients were assessed by clinical interview and examination, defecography, colon transit study, and anorectal manometry before randomization and 12 months postoperatively. Patients with compromised anal sphincter function or other symptomatic prolapse were excluded. RESULTS: The study groups were comparable in terms of demographic factors and rectocelerelated symptoms and signs. Eleven (73 percent) patients in the vaginal group and 10 (66 percent) in the transanal group digitally assisted rectal emptying preoperatively. The mean depth of the rectocele was 6.0 +/- 1.6 cm vs. 5.6 +/- 1.8 cm (P = 0.53) in the respective groups. At follow-up, 14 (93 percent) patients in the vaginal group and 11 (73 percent) in the transanal group reported improvement in symptoms (P = 0.08). Need to digitally assist rectal emptying decreased significantly in both groups, to one (7 percent) for the vaginal group and four (27 percent) for the transanal group (P = 0.17 between groups). The respective recurrence rates of rectocele were one (7 percent) vs. six (40 percent) (P = 0.04), and enterocele rates were zero vs. four (P = 0.05). In the vaginal group defecography showed a significant decrease in rectocele depth whereas in the transanal group the difference did not reach statistical significance. None of the patients reported de novo dyspareunia, but 27 percent reported improvement. CONCLUSION: Patients' symptoms were significantly alleviated by both operative techniques. The transanal technique was associated with more clinically diagnosed recurrences of rectocele and/or enterocele. Adverse effects on sexual life were avoided by use of both techniques. PMID- 15540293 TI - Patterns of fecal incontinence after anal surgery. AB - PURPOSE: Conservative anal surgery, with maximum preservation of the anal sphincters and continence, is becoming increasingly possible with the emergence of new sphincter-sparing treatments. Many surgeons remain skeptical, however, of the nature and impact of incontinence after anal surgery. We aimed to characterize the patterns of anal sphincter injury in patients with fecal incontinence after anal surgery. METHODS: We reviewed our fecal incontinence database and studied a subset developing incontinence after anal surgery. Maximum resting and squeeze pressures and the distal high-pressure zone to mid-anal canal resting pressure gradient were evaluated. Anal ultrasounds were evaluated and specific postoperative lesions were characterized. RESULTS: Patterns of sphincter injury in 93 patients with fecal incontinence after manual dilation, internal sphincterotomy, fistulotomy, and hemorrhoidectomy were studied. The internal sphincter was almost universally injured, in a pattern specific to the underlying procedure. One-third of patients had a related surgical external sphincter injury. Two-thirds of women had an unrelated obstetric external sphincter injury. The distal resting pressure was typically reduced, with reversal of the normal resting pressure gradient of the anal canal in 89 percent of patients. Maximum squeeze pressure was normal in 52 percent. CONCLUSION: Incontinence after anal surgery is characterized by the virtually universal presence of an internal sphincter injury, which is distal in the high-pressure zone, resulting in a reversal of the normal resting pressure gradient in the anal canal. These data support concerns that non-sphincter-sparing anal surgery leads to fecal incontinence and is increasingly difficult to justify given the availability of modern sphincter-sparing approaches. PMID- 15540294 TI - Sexual function following sphincteroplasty for women with third- and fourth degree perineal tears. AB - OBJECTIVE: Our goal was to evaluate sexual function following anal sphincteroplasty in women with third and fourth degree perineal tears secondary to birth trauma METHODS: Our study was performed using a retrospective cohort design in a group of women (n = 32) who had experienced either third-degree or fourth-degree perineal tears during labor and then elected to undergo sphincteroplasty for fecal incontinence. We surveyed our patients with a questionnaire that was developed by the Obstetrics and Gynecology Epidemiology Center at Harvard Medica School and was previously used to survey women with obstetric injuries. Self-reported presphincteroplasty and post sphincteroplasty degree of physical sensation, sexual satisfaction, and likelihood of achieving orgasm were measured Also measured were libido, partner satisfaction, and presence of emotional or physical inability to engage in sexual behavior. RESULTS: Our results reaffirmed the findings of the Obstetrics and Gynecology Epidemiology Center's study that sexual function is compromised in women with third and fourth-degree perineal tears. For our patients with this degree of perineal tearing who underwent sphincteroplasty after primary repair, our survey showed consistent improvement in several parameters of sexual function. After sphincteroplasty, physical sensation was higher/much higher in 40 percent, sexual satisfaction was better/much better in 33.3 percent, and 28.6 percent of the patients were more/much more likely to reach orgasm. Libido was improved in 37.5 percent of the study population, and 20 percent reported increased partner satisfaction. Before surgery, 23.5 percent of patients were physically and 31.2 percent emotionally unable to participate in sexual activity because of fear of incontinence or intimacy; after surgery only 6.3 percent were physically unable and 0 percent were emotionally unable to engage in sexual activity. The response rate for our study was 18/32 (56 percent). CONCLUSIONS: Anal sphincteroplasty for the treatment of incontinence in women with third- and fourth-degree perineal tears improves physical and emotional sexual well-being and function. PMID- 15540295 TI - Outcomes, risk of other malignancies, and need for formal mapping procedures in patients with perianal Bowen's disease. AB - PURPOSE: Local excision is standard treatment for Bowen's disease, but controversy exists over the incidence of synchronous and metachronous cancers as well as the role of a preexcision mapping procedure. METHODS: The medical records of 25 patients treated for perianal Bowen's disease between 1978 and 2001 were retrospectively reviewed. RESULTS: There were 11 men and 14 women with a mean age at diagnosis of 47.9 +/- 14.9 years. The majority of patients (72 percent) had symptom-driven biopsies. Presentation included pruritus (56 percent), mass (28 percent), bleeding (24 percent), and anal pain (4 percent). The average duration of symptoms before diagnosis was 1.3 +/- 0.5 years. Two patients had microscopic disease discovered incidentally after hemorrhoidectomy and underwent no further treatment. Twenty-three patients underwent wide local excision, of which 19 (83 percent) had a formal mapping procedure. On final pathology, 23 patients (92 percent) had clear margins. There were three recurrences (mean time to recur 2.0 +/- 1.0 years), including the two patients who had positive margins after wide excision (1 after mapping) and one patient who had clear margins after mapping. Five patients (20 percent) had other carcinomas (1 sigmoid, 4 vulvar). At follow up, 24 patients were alive with no evidence of Bowen's disease and 1 patient was dead secondary to sigmoid cancer with no evidence of Bowen's disease. CONCLUSIONS: Wide excision of perianal Bowen's disease results in excellent local control, although this is highly dependent on clear pathologic margins. A formal mapping procedure does not preclude recurrence. Initial screening and follow-up regimens are not uniform because of the wide range of treatment options available. PMID- 15540296 TI - APC genotype is not a prognostic factor in familial adenomatous polyposis patients with colorectal cancer. AB - PURPOSE: Several studies have shown that the clinical phenotype of patients with familial adenomatous polyposis is influenced by the position of the associated germline mutation in the APC gene. The aim of this work was to assess whether the site of the APC mutation may also predict the survival of familial adenomatous polyposis patients with a confirmed diagnosis of colorectal cancer. METHODS: A total of 387 familial adenomatous polyposis patients with colorectal cancer were examined. Of these, 287 (74 percent) belonged to families with an identified mutation, whereas 100 (26 percent) were from families in which no detectable APC mutation had been found by standard screening methods. The subjects were subdivided into four groups, according to the presence and localization of the identified mutation: with mutation before (a), at (b), or beyond codon 1309 (c), and without identified mutation (d). RESULTS: The cumulative five-year survival estimate of all cases included in the study was 0.56 (95 percent confidence interval, 0.51-0.61). No difference was observed in survival probability among patients from families with mutations before (0.56; 95 percent confidence interval, 0.49-0.63), at (0.58; 95 percent confidence interval, 0.43-0.72), or beyond (0.52; 95 percent confidence interval, 0.31-0.73) codon 1309 or those from families that were mutation negative (0.58; 95 percent confidence interval, 0.48 0.68) (log-rank test, P = 0.9). Survival analysis did not reveal any significant advantage for patients carrying a mutation in a specific region of the APC gene, after adjustment for age, gender, site, and stage. CONCLUSION: These data do not support the hypothesis that APC mutation may influence the outcome of familial adenomatous polyposis cases affected by colorectal cancer. PMID- 15540297 TI - Systemic and peritoneal angiogenic response after laparoscopic or conventional colon resection in cancer patients: a prospective, randomized trial. AB - PURPOSE: Angiogenesis is essential for wound healing. Vascular endothelial growth factor and endostatin are both endogenous angiogenic factors thought to be involved in the initiation and termination of angiogenesis. The aim of this study was to assess the local and systemic angiogenic profile in patients undergoing laparoscopic or open surgery for colon cancer. METHODS: Patients with primary colon carcinoma were prospectively randomized to curative laparoscopic (n = 12) or conventional (n = 14) resection. Vascular endothelial growth factor and endostatin levels in serum and wound fluid were investigated. RESULTS: In both groups vascular endothelial growth factor levels in wound fluid were significantly higher than postoperative serum levels, whereas endostatin levels in wound fluid were lower than serum levels and decreased progressively after surgery. The vascular endothelial growth factor levels in wound fluid measured at Day 4 were significantly higher in the laparoscopy group than in the laparotomy patients. CONCLUSIONS: Wound healing is associated with a strong local increase in pro-angiogenic factors and a decrease in antiangiogenic factors. The investigation of locally produced factors offered greater insight into the process of angiogenesis during wound healing than could be acquired from the circulation. PMID- 15540298 TI - A case-control study of laparoscopic right hemicolectomy vs. open right hemicolectomy. AB - PURPOSE: The purpose of our study was to examine all laparoscopic right hemicolectomies performed for cancer in our unit and to compare them with a case control series of open right hemicolectomies, with emphasis on long-term survival. METHODS: In a retrospective case-control series of right hemicolectomies, those done laparoscopically were compared with an age-matched and stage-matched series of patients who underwent open surgery. Survival was analyzed with the Kaplan-Meier method. RESULTS: Ninety-nine patients were included in the study, 33 laparoscopic and 66 open. Mean age 69.7 years. Dukes staging was the same between the two groups and mean follow-up period was 65.7 months. There were six laparoscopic conversions. The number of days patients were kept nil by mouth was significantly less in the laparoscopic cohort, with a mean of 2.4 days vs. a mean of 3.65 days (P = 0.005, Mann-Whitney U test). The number of days during which patients required parenteral opiates was significantly less in the laparoscopic cohort, with a mean number of days of 2.5, in contrast to 4.5 days in the open group (P = 0.008, Mann-Whitney U test). When overall survival was compared between the open and laparoscopic groups, no difference was found, with a mean overall survival of 40 months in the laparoscopic cohort and 39.4 months in the open cohort (P = 0.348, log-rank test). CONCLUSION: Laparoscopic right hemicolectomy for cancer does not compromise long-term survival and affords the advantage of a shorter period of postoperative ileus and decreased analgesia requirements. PMID- 15540299 TI - Does conversion of a laparoscopic colectomy adversely affect patient outcome? AB - PURPOSE: Conversion during laparoscopic colectomy varies in frequency according to the surgeon's experience and case selection. However, there remains concern that conversion is associated with increased morbidity and higher hospital costs. METHODS: From January 1999 to August 2002, 430 laparoscopic colectomies were performed by two surgeons, with 51 (12 percent) cases converted to open surgery. Converted cases were matched for operation and age to 51 open cases performed mostly by other colorectal surgeons from our department. Data collected included gender, American Society of Anesthesiology score, operative indication, resection type, operative stage at conversion, in-hospital complications, direct hospital costs, unexpected readmission within 30 days, and mortality. RESULTS: There were no significant differences between the groups for age (converted, 55 +/- 19; open, 62 +/- 16), male:female ratio (converted, 17:34; open, 23:28), or American Society of Anesthesiology score distribution. Indications for surgery were neoplasia (converted, 16; open, 31); diverticular disease (converted, 21; open, 13); Crohn's disease (converted, 12; open, 5); and other disease (converted, 2; open, 2). Operative times were similar (converted, 150 + 56 minutes; open, 132 +/ 48 minutes). Conversions occurred before defining the major vascular pedicle/ureter (50 percent), in relation to intracorporeal vascular ligation (15 percent), or during bowel transection or presacral dissection (35 percent). Specific indications for conversion were technical (41 percent), followed by adhesions (33 percent), phlegmon or abscess (23 percent), bleeding (6 percent), and failure to identify the ureter (6 percent). Median hospital stay was five days for both groups. In-hospital complications (converted 11.6 percent; open 8 percent), 30-day readmission rate (converted 13 percent vs. open 8 percent), and direct costs were similar between groups. There were no mortalities. CONCLUSION: Conversion of a laparoscopic colectomy does not result in inappropriately prolonged operative times, increased morbidity or length of stay, increased direct costs, or unexpected readmissions compared with similarly complex laparotomies. A policy of commencing most cases suitable for a laparoscopic approach laparoscopically offers patients the benefits of a laparoscopic colectomy without adversely affecting perioperative risks. PMID- 15540300 TI - Laparoscopic colorectal surgery modifies risk factors for postoperative morbidity. AB - PURPOSE: The aim of this study was to evaluate whether laparoscopic colorectal surgery can modify the risk factors for the occurrence of postoperative morbidity. METHODS: A total of 384 consecutive patients with colorectal disease were randomized to laparoscopic resection (n = 190) or open resection (n = 194). On admission, demographics, comorbidity, and nutritional status were recorded. Operative variables, patient outcome, and length of stay were also recorded. Postoperative complications were registered by four members of staff not involved in the study. RESULTS: The overall morbidity rate was 27.1 percent, with the rate in the laparoscopic group (18.7 percent) being less than that in the open group (31.5 percent; P = 0.003). Patients who underwent laparoscopic resection had a faster recovery of bowel function (P = 0.0001) and a shorter length of stay (P = 0.0001). In the whole cohort of patients, multivariate analysis identified open surgery (P = 0.003), duration of surgery (P = 0.01), and homologous blood transfusion (P = 0.01) as risk factors for postoperative morbidity. In the open group, blood loss (P = 0.01), homologous blood transfusion (P = 0.01), duration of surgery (P = 0.009), weight loss (P = 0.06), and age (P = 0.08) were related to postoperative morbidity. In the laparoscopic group the only risk factor identified was duration of surgery (P = 0.005). CONCLUSION: In the laparoscopic group, both postoperative morbidity and length of stay were significantly reduced and most risk factors for postoperative morbidity disappeared. PMID- 15540301 TI - Management of retained colorectal foreign bodies: predictors of operative intervention. AB - PURPOSE: This study was designed to review experience at our hospital with retained colorectal foreign bodies. METHODS: We reviewed the consultation records at Los Angeles County + University of Southern California General Hospital from October 1993 through October 2002. Ninety-three cases of transanally introduced, retained foreign bodies were identified in 87 patients. Data collected included patient demographics, extraction method, location, size and type of foreign body, and postextraction course. RESULTS: Of 93 cases reviewed, there were 87 individuals who presented with first-time episodes of having a retained colorectal foreign body. For these patients, bedside extraction was successful in 74 percent. Ultimately, 23 patients were taken to the operating room for removal of their foreign body. In total, 17 examinations under anesthesia and 8 laparotomies were performed (2 patients initially underwent an anesthetized examination before laparotomy). In the eight patients who underwent exploratory laparotomy, only one had successful delivery of the foreign object into the rectum for transanal extraction. The remainder required repair of perforated bowel or retrieval of the foreign body via a colotomy. In our review, a majority of cases had objects retained within the rectum; the rest were located in the sigmoid colon. Fifty-five percent of patients (6/11) presenting with a foreign body in the sigmoid colon required operative intervention vs. 24 percent of patients (17/70) with objects in their rectum (P = 0.04). CONCLUSIONS: This is the largest single institution series of retained colorectal foreign bodies. Although foreign objects located in the sigmoid colon can be retrieved at the bedside, these cases are more likely to require operative intervention. PMID- 15540302 TI - Effect of adenoviral-mediated transfer of transforming growth factor-beta1 on colonic anastomotic healing. AB - PURPOSE: Transforming growth factor-beta1 plays a central role in colonic repair. We examined the temporal effect of vector-mediated transfer of transforming growth factor-beta1 on colonic anastomotic healing. METHODS: Male Sprague-Dawley rats (n = 24) underwent transection of the distal colon and single-layer anastomosis. Proximal to the anastomosis, the colon was again transected and a colostomy was matured proximally. The distal colon was intubated with a silicone catheter, tunneled along subcutaneous tissues, and connected to a swivel apparatus for postoperative luminal infusion. Rats were randomized into four groups (n = 6 each). Two control groups received 10(10) plaque-forming units of a Type 5 E1-deleted adenovirus carrying the bacterial beta-galactosidase gene either immediately following surgery or on postoperative Day 3. The treatment groups received transforming growth factor-beta1 with the same viral construct at parallel time points. On postoperative Day 6, anastomotic bursting pressure and site were determined in situ with the anastomotic tissue subsequently harvested and analyzed by enzyme-linked immunosorbent assay for beta-galactosidase and transforming growth factor-beta1. RESULTS: When compared with its corresponding control, the group that received the transforming growth factor-beta1 gene on postoperative day 3 had a significantly higher bursting pressure (mmHg; 119 +/- 16 vs. 160 +/- 12, mean +/- SD; P = 0.001). While the majority of colons (5/6) from the control group burst at the anastomosis, none of the colons in the group that received transforming growth factor-beta1 on day 3 burst at the anastomotic site (P = 0.007). Beta-Galactosidase levels (pg/ml) in anastomotic tissue were significantly increased in both control groups when compared with their respective treatment groups (101 +/- 43 vs. 38 +/- 30, P = 0.01 when infused the day of surgery and 243 +/- 92 vs. 50 +/- 30, P = 0.009 when infused on day 3). Anastomotic levels of transforming growth factor-beta1 were also increased in the group receiving the transforming growth factor-beta1 gene on day 3 (214 +/- 66 vs. 135 +/- 24, P = 0.02). CONCLUSIONS: Gene transfer into the healing colonic anastomosis can be effectively achieved via intraluminal administration of adenoviral vectors. Transfer of transforming growth factor-beta1 increased the strength of colonic anastomoses when given at Day 3 but not at Day 0, demonstrating its diverse effects in the wound healing sequence. Thus, gene transfer of transforming growth factor-beta1 may avoid the need for a diverting stoma in cases of rectal surgery and impaired healing resulting from chemotherapy or radiation. PMID- 15540303 TI - Irrigation volume determines the efficacy of "rectal washout". AB - PURPOSE: Rectal stump washout has been recommended to prevent implantation of exfoliated malignant cells in the anastomosis after anterior resection for rectal cancer. The aim of this study was to investigate its efficacy, particularly the extent to which the volume of irrigation fluid might influence the efficacy of tumor cell elimination and whether tumor characteristics might influence the result. METHODS: The study comprised 30 consecutive patients operated on by anterior resection for rectal cancer. After cross-clamping the rectum below the tumor, a washout sample was collected for examination after every incremental 500 ml of saline irrigation up to 2 liters. The presence of shed cancer cells was correlated with the washout volume and tumor characteristics. RESULTS: Cancer cells were found in 29 of 30 patients (97 percent) in the first sample of irrigation fluid and decreased gradually in frequency and number with increasing irrigation volumes. No cancer cells were demonstrated after 1.5 liters of irrigation in patients with tumor below the peritoneal reflection, whereas cancer cells were still present in one-fourth of the patients with tumor located above the peritoneal reflection. Finally, only a small number of cancer cells was confirmed in one patient after 2 liters of irrigation. CONCLUSIONS: The irrigation volume determined the efficacy of rectal washout. With our method, 1 1/2 liters of saline irrigation appears to clear contents from cancer cells in patients with tumors below the peritoneal reflection whereas at least 2 liters is recommended for patients with tumor above the peritoneal reflection. PMID- 15540304 TI - Dynamic graciloplasty in patients born with an anorectal malformation. AB - PURPOSE: The aim of this study was to compare long-term results for patients born with an anorectal malformation and fecal incontinence treated with a dynamic graciloplasty with those for the total group of patients undergoing dynamic graciloplasty. METHODS: Consecutive patients with fecal incontinence after surgical treatment of anorectal malformation and treated with dynamic graciloplasty were included in this study. Preoperative assessment was performed. Postoperative follow-up consisted of anorectal manometry and registration of defecation frequency, continence scores, and postponement time of defecation. RESULTS: Twenty-eight patients with a median age of 25.5 years were included in the study. The median follow-up was 4 years. A high anorectal malformation was present in 89.3 percent of patients. Conventional graciloplasty had been previously performed in 36 percent. All patients were incontinent for stools. Median frequency of defecation was four times/day. Median postponement time of defecation was 0 minutes. Rectoanal inhibition reflex was present in 17 percent of patients. Median preoperative sensory threshold during balloon distention was 30 ml and median maximum urge threshold was 165 ml. Satisfactory continence was reached in 35 percent of patients, however, 7.1 percent of patients gained this continence score by additional bowel irrigation. Twenty-nine percent of patients were incontinent for loose stool, 36 percent were incontinent for formed stool. Satisfactory continence was achieved in only 18 percent of patients with a high anorectal malformation, compared with 100 percent in patients with a low anorectal malformation. In the total group of patients with dynamic graciloplasty, satisfactory continence was obtained in 76 percent. The sensitivity threshold in patients with a successful dynamic graciloplasty was lower than that in patients with a failing dynamic graciloplasty (45 vs. 24 ml, P = 0,06). When we compare median preoperative rectal sensitivity threshold in our study group with that in the total patient group with dynamic graciloplasty, statistical difference was established (P = 0.008). Postponement time (0 to 20 minutes) and anal squeeze pressure (81 to 120 mmHg) increased significantly after surgery. Patients with an anorectal malformation had significantly lower resting and stimulation pressure than that of the total group of patients, but the difference between resting and stimulation pressure in both groups was not significantly different (P = 0.33). The difference between resting and stimulation pressure was not significantly different between anorectal malformation patients with a failing dynamic graciloplasty and patients with a successful dynamic graciloplasty. Complications were noted in 57 percent of patients. Explantation of the dynamic graciloplasty was necessary in 32 percent of patients, mainly because of infection of the implant. CONCLUSIONS: Results of dynamic graciloplasty for fecal incontinence are reasonable for this specific group of patients with limited treatment options. Despite functional dynamic graciloplasty, the results are worse than those for the total group of patients with dynamic graciloplasty. Rectal sensitivity and type of malformation are prognostic factors for outcome and can be used to select patients for treatment with dynamic graciloplasty, thereby improving treatment outcome. PMID- 15540305 TI - Innovations in fecal incontinence: sacral nerve stimulation. AB - OBJECTIVE: The aim of this study was to present an overview of sacral nerve stimulation in the treatment of fecal incontinence. We describe the evolution in technique, patient selection, and indications, and review results and complications. METHODS: All articles on sacral nerve stimulation for fecal incontinence that were recovered on MEDLINE search were reviewed. With multiple articles from an institution, the most recent reports with the longest follow-up and largest cohort of patients were selected, unless information from earlier reports was relevant. RESULTS: The technique of sacral stimulation is well established, carries little risk, and continues to be refined (e.g., a less invasive approach has been proposed). Patient selection is based on a two-stage diagnostic test stimulation (acute and subchronic), for which the predictive value is high. On this basis, permanent sacral nerve stimulation has proved effective in both single-center and multicenter trials in patients with a functional deficit but limited morphologic lesions or no morphologic lesions. The clinical benefit derives from multiple symptomatic improvements contributing to better bowel control and from substantially improved quality of life. The underlying mechanism of action remains undefined, but both somatic and autonomic function appears affected. CONCLUSION: Sacral nerve stimulation offers a safe treatment mode in a patient population in whom conservative treatment has failed and traditional surgical approaches would have limited success. The high predictive value of the diagnostic approach offers a unique therapeutic advantage. PMID- 15540306 TI - Fournier's gangrene after Milligan-Morgan hemorrhoidectomy requiring subsequent abdominoperineal resection of the rectum: report of a case. AB - The development of Fournier's gangrene after Milligan-Morgan hemorrhoidectomy of a previously healthy 76-year-old female patient is described. After such a common surgical procedure, the patient developed full-thickness skin necrosis of the perianal region including the rectum. Immediate radical debridement was mandatory. Because of rectal involvement, a diverting sigmoid colostomy was required. The rectum had to be removed by abdominoperineal resection. This disastrous complication was completely unexpected and unpredictable after Milligan-Morgan hemorrhoidectomy because of the lack of predisposing factors. PMID- 15540307 TI - First description of a PEComa (perivascular epithelioid cell tumor) of the colon: report of a case and review of the literature. AB - We describe a young female patient suffering from a PEComa (perivascular epithelioid cell tumor) of the cecum, incidentally found at an examination made by her family physician. The perivascular epithelioid cell tumor is a very rare tumor, until today reported in a few cases in falciform ligament, uterus, jejunum, terminal ileum, rectum, liver, kidney, lung, pancreas, prostate, and soft tissue of the thigh. This tumor is part of a new group of tumors, comprised of angiomyolipoma, lymphangiomyolipoma, and clear-cell myomelanocytic "sugar" tumor. Defined by coexpression of melanocytic (HMB-45) and muscle markers (smooth muscle actin and desmin) the perivascular epithelioid cell tumor does not have predictable histopathologic behavior. Some cases of metastasis are described, comorbidities such as tuberous sclerosis of the brain "Bourneville" and lymphangioleiomyomatosis have to be excluded. The therapy consists of the radical resection. An adjuvant therapy is not known. Recommended is a close and long-term follow-up clinically and by CT scan. PMID- 15540308 TI - Colonic obstruction caused by adhesive splenomegaly: report of a case. AB - INTRODUCTION: Splenomegaly caused by hematologic or infectious diseases is a well known complication that may cause discomfort because of increased intra-abdominal pressure; however, complete obstruction of the large bowel usually never occurs. METHODS: We present a case of a 76-year-old male with polycythemia vera admitted with symptoms of complete colonic obstruction. At operation, the descending colon was found to be completely obstructed by a large adhesive spleen. Incision of the adhesions, complete colonoscopy, and mobilization of the colon was performed. RESULTS: Afterward, the patient had a normal colonic function; however, four days later he had a magnetic resonance-confirmed cerebral thrombosis followed by an episode of aspiration pneumonia, which he died of ten days after the operation. CONCLUSIONS: Colonic obstruction caused by splenomegaly is extremely rare, but may happen in patients with hematologic diseases. PMID- 15540309 TI - H-pouch: new isoperistaltic colonic pouch for coloanal anastomosis after rectal resection for cancer: a pilot study. AB - PURPOSE: To date, there is a consensus to consider colonic J-pouch-anal anastomosis as the standard form of reconstruction after proctectomy for cancer. However, one drawback of colonic J-pouch is evacuation difficulties, which are observed at some degree in almost one-half of patients. To improve functional outcome after coloanal anastomosis, we describe a new technique of isoperistaltic colonic H-pouch. METHODS: The functional results after construction of this pouch were assessed prospectively in ten patients and compared with those obtained in a control group of matched patients with a colonic J-pouch. RESULTS: At six months and one year, there were no differences between both pouches for bowel function, continence score, and evacuation difficulties. CONCLUSIONS: This preliminary study showed that the creation of a new isoperistaltic colonic H-pouch did not improve the functional results after coloanal anastomosis compared with colonic J pouch. Because colonic H-pouch is technically more complex to fashion, it should not be performed routinely and the J-pouch remains the benchmark for routine clinical practice. PMID- 15540310 TI - Vacuum-assisted closure of complex perineal wounds. AB - The Vacuum-assisted Closure device decreases the time to wound healing, thus increasing the deposition of granulation tissue, and decreasing the use of wound care specialists. Perineal wounds present a special challenge. We present four cases of complex perineal wounds in which the Vacuum-assisted Closure device was used. In each case, wound care was simplified and healing accelerated. The Vacuum assisted Closure device allows earlier wound closure, early skin grafting (with improved graft adherence), earlier hospital discharge, and earlier return to baseline functional status. Its use in the perineum presents a challenge, but with proper application, even the most complex perineal wounds can be healed. PMID- 15540311 TI - Primary sphincter repair: are the results of the operation good enough? PMID- 15540312 TI - A world of colorectal surgery. PMID- 15540313 TI - Meeting report from the XV International AIDS Conference. Access in the developing world. PMID- 15540315 TI - Meeting report from the XV International AIDS Conference. Clinical trials of approved antiretrovirals. PMID- 15540314 TI - Meeting report from the XV International AIDS Conference. Perinatal transmission. PMID- 15540316 TI - Meeting report from the XV International AIDS Conference. New drugs. PMID- 15540317 TI - Meeting report from the XV International AIDS Conference. Structured treatment interruption and other novel treatment strategies. PMID- 15540318 TI - Meeting report from the XV International AIDS Conference. Metabolic complications. PMID- 15540319 TI - Meeting report from the XV International AIDS Conference. Opportunistic infections. PMID- 15540320 TI - Meeting report from the XV International AIDS Conference. Adherence. PMID- 15540321 TI - Meeting report from the XV International AIDS Conference. Prevention. PMID- 15540322 TI - Human rights and HIV/AIDS in the context of 3 by 5: time for new directions? AB - Over the last decade, the success of the human rights-based approach to HIV/AIDS has been spotty, says Mark Heywood. In this feature article, the author describes the challenges that remain in implementing a human rights approach. He presents an analysis of questions raised by De Cock et al concerning the applicability of the human rights approach. The author argues that human rights advocacy needs to continue, but that new directions are required. The article outlines new directions in the areas of (a) confidentiality and openness, (b) HIV testing, and (c) health systems. The author concludes that the most serious threat to human rights remains the unwillingness of national governments to take all necessary measures to build health services and prevent epidemics. PMID- 15540323 TI - About harm reduction in the context of the response to HIV/AIDS. PMID- 15540324 TI - Harm-reduction activism: a case study of an unsanctioned user-run safe injection site. AB - Due to the ongoing health crisis among injection drug users in Vancouver, Canada, there have been repeated calls for the establishment of safe injection sites (SISs) since the early 1990s. In April 2003, in response to a large-scale police crackdown and government inaction, a group of activists opened an unsanctioned SIS in Vancouver's Downtown Eastside (DTES). The 327 Carrall Street SIS operated for 181 days despite considerable police harassment and limited financial support. During the operation of the SIS, volunteers supervised over 3000 injections and demonstrated the feasibility of a user-run low-threshold SIS. The experience of the SIS provides valuable lessons for those seeking to advance the interests of injection drug users through community mobilization and direct action approaches. In this article, Thomas Kerr, Megan Oleson, and Evan Wood describe the events surrounding the establishment, operation, and closing of the unsanctioned SIS, and outline the lessons learned. PMID- 15540325 TI - Canada announces new funding for domestic and global fight against AIDS. AB - In a flurry of announcements that came just weeks before it called a federal election, the federal government doubled the funding for its domestic HIV/AIDS strategy, doubled its contribution to the Global Fund to Fight AIDS, Tuberculosis and Malaria, and announced a significant contribution to the World Health Organization (WHO) 3 by 5 Initiative. AIDS organizations welcomed the announcements but were critical of the fact that the increased funding for the domestic strategy will take five years to implement. They also criticized the fact that funding for the global initiatives is not new money; it will come out of increases for development assistance previously announced. PMID- 15540326 TI - Reports call for end to harms caused by Criminal Code prohibitions surrounding prostitution. AB - The sale of sexual services between consenting adults is legal under Canadian law, yet the Criminal Code prohibits many activities related to prostitution and the sex trade. As a result, many people who work in the sex trade come into conflict with the law. In recent reports, two Canadian organizations have called for the repeal of the current Criminal Code prohibitions on activities related to the sex trade. Pivot Legal Society's report, "Voices for Dignity: A Call to End the Harms Caused by Canada's Sex Trade Laws," was released in March 2004. The Conseil permanent de la jeunesse, a 15-member advisory panel to the Quebec government, released "Prostitution de rue: avis" in April 2004. PMID- 15540327 TI - Nova Scotia renews its HIV/AIDS strategy. AB - On 1 December 2003, after a five-year process of consultation and planning, Nova Scotia embarked on a new HIV/AIDS strategy. It replaces the first strategy, launched a decade earlier. The renewed strategy is meant to promote collaborative action on the determinants of vulnerability to HIV infection and on the capacity of people living with HIV/AIDS to achieve optimal health and quality of life. PMID- 15540328 TI - Study highlights negative public health consequences of police crackdown on drug market. AB - A police crackdown in Vancouver's Downtown Eastside (DTES) displaced the drug scene to adjacent areas and may have exacerbated the health and social harms associated with injection drug use. This is the finding of a study published in the Canadian Medical Association Journal by Evan Wood and his colleagues at the British Columbia Centre for Excellence in HIV/AIDS. PMID- 15540329 TI - Health Canada considers dispensing medical marijuana through pharmacies. AB - Health Canada may be poised to emulate the Netherlands' system of distributing marijuana to HIV/AIDS and other patients through pharmacies. Meanwhile, revisions to the much-criticized medical marijuana regulatory system are under development. PMID- 15540330 TI - Montreal: probe requested into police hiring practices. PMID- 15540331 TI - Quebec: College issues statement on HIV testing for physicians. PMID- 15540332 TI - Statistics Canada plans to use blood and other samples to measure health trends. PMID- 15540333 TI - Alberta: "Blood Samples" act passes third reading. PMID- 15540334 TI - International news--10 years on. PMID- 15540335 TI - Ghana: capacity-building workshops for lawyers and judges. AB - In 2004, AIDS Alert Ghana (an Accra-based NGO) commenced a series of two-day workshops to enhance the capacity of lawyers and judges to respond to HIV/AIDS. The workshops are organized by the AIDS Alert Law Project (AALP). PMID- 15540336 TI - Comparative study of HIV/AIDS and human rights in Andean nations. AB - A comparative study of five nations of the Andean community has revealed serious gaps in meeting international commitments to address HIV/AIDS-related human rights. PMID- 15540337 TI - Report documents human rights abuses of drug users in Thailand. AB - A violent, state-sponsored "war on drugs" is jeopardizing Thailand's long struggle to become one of Southeast Asia's leading rights-respected democracies. This is one of the findings of a report released by Human Rights Watch (HRW) on the eve of the XV International AIDS Conference in Bangkok, Thailand in July 2004. PMID- 15540338 TI - The Philippines: HIV/AIDS, condoms, and human rights. AB - Influential leaders are increasingly attacking condoms through HIV-prevention programs that focus on sexual abstinence and marital fidelity. In May 2004 Human Rights Watch (HRW) released a report that examines the impact of these policies in the Philippines, the largest Roman Catholic country in Asia and a major recipient of US HIV/AIDS funding. PMID- 15540339 TI - Russian Federation: Battle not over in drug-law changes. AB - As reported in the last issue of the Review, in late 2003 the Russian Parliament amended the national drug law to allow for the possibility of less harsh penalties for small-scale drug users. The Parliament left the development of accompanying regulations to the federal drug-control authorities and the Ministry of Health. These two entities were meant to recommend new levels of minimum dosages that would qualify users for criminal penalties if those amounts were found in their possession. Rather than acting in the spirit of the Parliament's amendment, in March 2004 the drug-control authorities circulated a proposal for minimum dosages so tiny that virtually any level of possession would incur a long prison sentence. PMID- 15540340 TI - Hungary: a fight every step of the way for voluntary, anonymous, and free tests. AB - Voluntary, anonymous, and free HIV tests must be addressed in developing any effective policy in the fight against HIV/AIDS. There are heated discussions about the first two criteria, particularly anonymous testing, but free testing can also become contentious, as the following story shows. PMID- 15540341 TI - USA: Cirque du Soleil pays US$600,000 to end HIV discrimination complaint. AB - In April 2004 Lambda Legal, a US-based civil rights organization, announced that the travelling circus Cirque du Soleil would pay a record US$600,000 to end an HIV discrimination complaint filed by one of its former employees who was dismissed because he has HIV. PMID- 15540342 TI - Namibia: policy adopted on HIV/AIDS and the education sector. AB - The Namibian National Policy on HIV/AIDS for the Education Sector, adopted in 2003, provides a framework for prevention, care, and support for both learners and employees in the education sector and for the mitigation of the impact of HIV/AIDS on learners and employees, as well as on the education sector itself. PMID- 15540343 TI - Global: HIV/AIDS-related travel restrictions--UNAIDS/IOM statement and revision of International Health Regulations. AB - In June 2004 UNAIDS and the International Organization for Migration (IOM) issued a joint statement on HIV/AIDS-related travel restrictions. While acknowledging that the regulation of immigration matters is widely recognized as falling within the sovereign power of the individual state concerned, the statement distinguishes between short-term (one month or less) and long-term travel. PMID- 15540344 TI - Legal and ethical issues in microbicides research and development in Canada. AB - The Canadian AIDS Society (CAS) recently completed a report entitled Microbicides Development and Delivery in Canada: Legal, Ethical and Human Rights Issues. The report builds on Canadian and international experience and was written in consultation with Canadian community and international experts. It is available on the CAS website (www.cdnaids.ca) and from the Canadian HIV/AIDS Information Centre (www.aidssida.cpha.ca) as of September 2004. In this article the report's author, Anna Alexandrova, argues that Canada needs to develop a microbicides development and delivery strategy that addresses research and development issues, outlines possible roles for meaningful community participation, and provides guidelines on funding, promotion, licensing, and distribution. PMID- 15540345 TI - Consumer groups call for ban on internet pharmacies. AB - A number of consumer groups have called on the Canadian government to ban internet pharmacies, claiming that the industry is putting the health of Canadians at risk. The groups say that the impact will escalate if the problem is not addressed. PMID- 15540346 TI - Therapeutic HIV vaccine trial launched in Canada. AB - The first-ever clinical trial of a therapeutic HIV vaccine in Canada is currently underway. The trial is being run by the Canadian Network for Vaccines and Immunotherapeutics (CANVAC) in cooperation with Aventis Pasteur, of France, and the Immune Response Corporation (IRC), of the United States. PMID- 15540347 TI - Correctional Service Canada to undertake Safer Tattooing Practices Initiative. AB - In 1994, the Expert Committee on AIDS and Prisons recommended that tattooing equipment and supplies be authorized for use in federal correctional institutions, and that prisoners who would offer tattooing services to other prisoners be instructed on how to use tattooing equipment safely. Ten years later, Correctional Service Canada (CSC) has finally announced that, as part of a Safer Tattooing Practices Initiative, it will set up safer tattooing pilot projects in six federal prisons in 2004, and evaluate the initiative. PMID- 15540348 TI - Health care needs assessment of federal prisoners in Canada released. AB - On 7 April 2004, Correctional Services Canada (CSC) released a comprehensive profile of the health needs of federal prisoners in Canada, published in a special supplement to the Canadian Journal of Public Health. PMID- 15540349 TI - United Nations agencies release policy brief on reduction of HIV transmission in prisons. PMID- 15540350 TI - US Judge: inadequate medical care for HIV-positive prisoners is a violation of rights. AB - A federal judge in the United States has determined that Alabama prison officials violated HIV-positive prisoners' constitutional rights and that poor medical care caused HIV-positive prisoners to die early. PMID- 15540351 TI - Criminal law and HIV transmission/exposure: three new cases. PMID- 15540352 TI - Fear of persecution not adequate to claim refugee status. AB - In Delgadillo v Canada, the Federal Court of Canada upheld an Immigration and Refugee Board (IRB) decision to refuse refugee status to a Mexican gay male refugee claimant because he had an Internal Flight Alternative (IFA). Although the court reached this conclusion, it nevertheless agreed that Mr Delgadillo had a legitimate fear of persecution. As well, the court made an important finding- that people in Mexico who are living with HIV/AIDS do not have an IFA. PMID- 15540353 TI - US: immigration court allows Zambian woman's asylum claim based on HIV infection. AB - On 9 February 2004, the federal Immigration Court ruled that a Zambian woman can stay in the US because her HIV-positive status gives her a well-founded fear of persecution in Zambia. The court found that she would face severe and lethal discrimination in the public health clinics and in the employment sector and that the Zambian government is unwilling or unable to control this persecution. PMID- 15540354 TI - Australia: Migration Review Tribunal waives medical inadmissibility criteria for two HIV-positive visa applicants. AB - In two recent cases, Australia's Migration Review Tribunal (MRT) overturned government decisions to refuse the visa applications of HIV-positive candidates for Partner (Temporary) (Class UK) visas on the basis of medical inadmissibility. In doing so, the MRT questioned the opinion of the government medical officer concerning the costs to the Australian community, and took into account the applicants' potential contributions. PMID- 15540355 TI - Botswana: Industrial Court tackles HIV testing in the workplace. AB - The Botswana Industrial Court recently decided two cases regarding mandatory HIV testing in the workplace. One case addressed constitutional rights of HIV infected people, expanding the reach of the Bill of Rights to the private sphere and potentially offering wide protection to people living with HIV/AIDS. Both cases highlight the glaring need for HIV-related legislation in Botswana. PMID- 15540356 TI - India: constitutional protection from discrimination in employment on the basis of HIV status affirmed in three cases. PMID- 15540357 TI - UK: Court of Appeal orders retrial in UK's first HIV criminal transmission case. AB - On 5 May 2004, the UK Court of Appeal, Criminal Division, granted Mohammed Dica's appeal against his October 2003 conviction on two counts of causing grievous bodily harm for reckless transmission of HIV. The court ordered a retrial after determining that the trial judge was wrong to not allow the defendant to present information that the victims had known about his condition and had consented to the risk. PMID- 15540358 TI - Australia: Supreme Court of Western Australia orders retrial in case of man convicted of infecting girlfriend with HIV. AB - On 3 October 2002, the District Court convicted an HIV-positive man, Houghton, of unlawfully causing grievous bodily harm to his girlfriend for having unprotected vaginal and anal intercourse with her. Houghton had not told her of his HIV status prior to intercourse. He had been aware of his status for some years before meeting the woman, but testified at his trial that he believed that he could not transmit HIV if he did not ejaculate inside the woman. The woman became HIV-positive as a result of the sexual intercourse. The question put before the jury was twofold: whether there had been bodily injury caused by the applicant to the complainant; and whether that injury was of sufficient severity to constitute grievous bodily harm. The jury found Houghton guilty and in doing so made a finding that the transmission of HIV constitutes grievous bodily harm. PMID- 15540359 TI - Treatment Action Campaign (TAC) v South Africa (Minister of Health): reflections on the right to health care in Canada. AB - In this article, Showkat Yazdanian examines the potential impact of the South African Constitutional court's decision in TAC v Minister of Health on the right to health care in Canada. Showkat first focuses on the potential utility of international treaties and precedents as a means to uphold the right to preventative health care in Canada. She then examines the Canadian Constitution's bearing on a right to health care, including an analysis of the current division of federal and provincial health powers. PMID- 15540360 TI - Britain grants license to make human embryos for stem cells. PMID- 15540361 TI - Tubes, pump and fragile hope keep a baby's heart beating. PMID- 15540362 TI - Britain embraces embryonic stem cell research. PMID- 15540363 TI - U.S. Court in New York rejects partial-birth abortion ban. PMID- 15540364 TI - Glaxo agrees to post results of drug trials on web site. PMID- 15540365 TI - Identity chip planted under skin approved for use in health care. PMID- 15540366 TI - US creates ethics panel on priority for flu shots. PMID- 15540368 TI - Buying charts. PMID- 15540367 TI - You don't have to go down with the ship. PMID- 15540369 TI - Retraction of "cloning and analysis of the rat glutamate-cysteine ligase modifier subunit promoter" [Biochem. Biophys. Res. Commun. 285(2001):476-482]. PMID- 15540370 TI - Patients' preferences and experiences in handling emotions: a study on communication sequences in primary care medical visits. PMID- 15540371 TI - Modern histamine H1-receptor antagonists in the unified airway. PMID- 15540372 TI - Medical schools adopt new research rules. PMID- 15540373 TI - Making house calls wherever the patient calls home. PMID- 15540374 TI - Privacy concerns spur plan to review questionnaire for medical school graduates. PMID- 15540375 TI - The politics of bioethics. PMID- 15540376 TI - Roe revisited: a grim fairy tale. PMID- 15540378 TI - Cosmetic surgery in halacha. PMID- 15540377 TI - A boy or a girl? The ethics of preconception gender selection. PMID- 15540379 TI - Origin and development of the Tri-Council Policy Statement on the Ethics of Research Involving Humans. PMID- 15540380 TI - NCEHR workshop: institutional response. PMID- 15540381 TI - Research ethics boards--operational issues I. PMID- 15540382 TI - Research ethics boards--operational issues II. PMID- 15540383 TI - The analysis of risks and potential benefits in research. PMID- 15540384 TI - Process and context--ethics review of research impacting aboriginal and indigenous peoples: establishing the historical context. Introduction to the Proceedings of a Workshop of the Consent Committee, November 19-21, 1999 held in Ottawa, Canada. PMID- 15540385 TI - Community representation: broadening the perspective and value base of research ethics boards. PMID- 15540386 TI - Minimal risk and its implications. Proceedings of the NCEHR National Workshop on Minimal Risk, March 31-April 1, 2000 held in Ottawa, Ontario, Canada. PMID- 15540387 TI - New horizons in the development of antiepileptic drugs: the search for new targets. AB - The past decades have brought many advances to the treatment of epilepsy. However, despite the continued development and release of new antiepileptic drugs, many patients have seizures that do not respond to drug therapy or have related side effects that preclude continued use. Even in patients in whom pharmacotherapy is efficacious, current antiepileptic drugs do not seem to affect the progression or underlying natural history of epilepsy. Furthermore, there is currently no drug available which prevents the development of epilepsy, e.g., after head trauma or stroke. Thus, there are at least three important goals for the future: (1) better understanding of processes leading to epilepsy, thus allowing to create therapies aimed at the prevention of epilepsy in patients at risk; (2) development of disease-modifying therapies, interfering with progression of epilepsy, and (3) improved understanding of neurobiological mechanisms of pharmacoresistance, allowing to develop drugs for reversal or prevention of drug resistance. The Second Workshop on New Horizons in the Development of Antiepileptic Drugs explored these three goals for improved epilepsy therapy, with a focus on the search for new drug targets for prevention of epilepsy, for interfering with progression of epilepsy, and for interfering with drug resistance in epilepsy. A special topic dealt with gene expression analysis for target identification. Furthermore, pharmacological and non pharmacological targets for curing epilepsy were explored. In this conference review, the current status of antiepileptic therapies is critically assessed, and innovative approaches for future therapies are highlighted. PMID- 15540388 TI - Protocols for the reporting of cutaneous melanoma. PMID- 15540389 TI - Checklists and guidelines in surgical pathology. PMID- 15540390 TI - "Do this, it could save your life!" and other problematic claims in preventive medicine. PMID- 15540391 TI - Genetic ancestry tracing and American Indian identity. PMID- 15540392 TI - The co-evolution of bioethics, computing and cyberspace: an archeological perspective. PMID- 15540393 TI - Internet censorship in the hospital: bad ethics and great irony. PMID- 15540394 TI - The correct Caduceus. PMID- 15540395 TI - Addressing the effects of adverse events: study provides insights into patient safety at Canadian hospitals. PMID- 15540396 TI - Volume and outcome in healthcare: implications for health policy. PMID- 15540397 TI - In-hospital hip fractures in Canada: using information to improve patient safety. PMID- 15540398 TI - Securing the future of Canada's Academic Health Sciences Centres. AB - Although there have been a number of in-depth national, federal, provincial and territorial reviews of the health system in Canada, there has not been a systematic review of the current mission/mandate and future roles and responsibilities of Canada's Academic Health Sciences Centres (AHSCs) since the early 1990s--this despite the fact that the system has experienced profound change. With this in mind, more than 20 national health organizations and others have engaged in discussions about the importance of establishing a national task force to consider and develop recommendations that will focus on the future role of Canada's AHSCs. PMID- 15540399 TI - Aligning the stars: Using systems thinking to (re)design Canadian healthcare. AB - It has become commonplace to refer to a "Canadian healthcare system," implying some monolithic system of prevention and care, yet failing to acknowledge its various subsystems, some performing well and some not. This article argues that a more nuanced understanding of healthcare systems, based on the principles of Systems Theory, provides a useful lens to understand our current system(s) as well as those that are possible, the principles of design and the levers available to leaders and policymakers as we implement our healthcare strategies for Canada. PMID- 15540400 TI - Aim for excellence: integrating accreditation standards into the continuous quality improvement framework. AB - The Ottawa Hospital has developed a simple template incorporating specific key Canadian Council on Health Services Accreditation criteria that will allow clinical teams to annually self-assess their activities against the accreditation standards. PMID- 15540401 TI - "What's in, what's out": stakeholders' views about the boundaries of Medicare. AB - The Canada Health Act requires that provincial insurance plans provide universal coverage without co-payments for all "medically necessary" services delivered by hospitals and doctors, but allows care delivered by other providers in other locations to fall outside of the boundaries of Medicare. Discussion about the sustainability of medicare at both the national and provincial levels has called for the revisiting of these boundaries. The M-THAC (Medicare to Home and Community) Research Unit attempted to clarify the areas of consensus and controversy as to what key stakeholders thought should be "in" or "out" of Medicare. Using a non-experimental, cross-sectional design, a self-administered survey (in both English and French, constructed in consultation with our partners) was distributed between January and April 2002 to policy elites of key stakeholder groups. The results are based on 2,523 responses. Much of the current "debate" is mired in discussing issues where consensus already exists. We found strong support for in-hospital care. However, there is considerable resistance, across all groups, to full funding for similar services in private clinics or in the home, and almost no support for full funding for non-medical home-based services. The vision of many policy elites remains heavily linked to the current system of guaranteed public funding only for acute care in hospitals or by physicians. Successful reform will need to address, rather than assume, a broader view of healthcare. PMID- 15540402 TI - Who's the fairest of them all? Which provincial pharmacare model would best protect Canadians against catastrophic drug costs? AB - BACKGROUND: Public prescription drug plans vary markedly across Canada. To address perceived inequities in coverage across provinces, the February 2003 First Ministers' Accord on Health Care Renewal committed to ensuring that all Canadians have reasonable access to catastrophic drug coverage. A national standard for "reasonable" catastrophic coverage has yet to be formally defined. OBJECTIVE: To compare the private financial burdens from prescription drugs that Canadian households would face if each of the current provincial pharmacare models were adopted as the national standard. METHODS: Through simulation modelling, we computed household private financial burden by applying the cost sharing rules from provincial drug plans to a nationally representative set of 4,860 household types differing in size, age composition, income and drug expense levels. The proportions of households that would face private out-of-pocket payments exceeding critical, or catastrophic, percentages of household income were calculated. RESULTS: Private financial burden due to prescription drug costs varies considerably across provincial pharmacare models. Comprehensive, tax financed pharmacare models that limit out-of-pocket expenditures to a given percentage of income, such as those found in British Columbia, Saskatchewan, Manitoba and Ontario, provide the greatest protection against catastrophic prescription drug costs. There appears, however, to be no "gold standard" for an acceptable financial burden to be borne by patients. PMID- 15540403 TI - Family physicians and home care agencies--valuing each other's roles in primary care. AB - BACKGROUND: In this era of primary-care reform, family physicians are being encouraged to work in teams with allied health professionals and community resources, including home care services, to ensure optimum care is provided to their patients. OBJECTIVE: To learn about the experiences of physicians working with home care services as provided by the Toronto Community Care Access Centre. METHODS: In early 2001, the Toronto Community Care Access Centre (CCAC) hosted focus groups designed to understand physicians' knowledge of and experiences with the home care services provided by our organization. Data analysis was conducted using grounded theory methodology. RESULTS: Three themes emerged: (1) family physicians have a limited understanding of home care services; (2) family physicians felt there were inconsistencies related to service provision by CCAC staff; and (3) family physicians felt that their role may not be valued when caring for mutual clients with CCACs. OUTCOMES: Better educational interventions informing physicians on CCAC services and improved organizational practices by CCACs may aid in improving the connection between family physicians and CCACs. PMID- 15540405 TI - Hospital redevelopment: crisis, challenge or opportunity! One rural hospital's success story. AB - Listowel Memorial is a 50-bed acute care facility located in southwestern Ontario, serving a rural population of close to 30,000 across a broad geographical area. It is currently in the midst of a dollars 15 million phased redevelopment and renewal. PMID- 15540406 TI - Coming soon to a health sector near you: An advance look at the new Ontario Personal Health Information Protection Act (PHIPA). AB - This is Part I of a two-part article that provides a broad overview of the new health sector privacy legislation in Ontario, and compares this legislation to personal health legislation in other provinces. In Part I, we discuss the objectives, structure and scope of, as well as the substantive rights and obligations created by, the Ontario Act. In Part II, which will appear in the Fall 2004 issue, we will discuss the administrative obligations under the Ontario Act, as well as the provisions relating to enforcement and remedies. We will also discuss the approach to the protection of personal health information taken by other provinces, including Alberta, Saskatchewan and Manitoba, which have already enacted legislation that is similar in many respects to the Ontario Act. PMID- 15540404 TI - The impact of allocation of additional resources on the waiting time for cataract surgery. AB - In 2001, the ophthalmology community in Manitoba lobbied the regional health authority to allocate additional resources for cataract surgery because of unacceptably long waits. Approval was given in principle for a partial increase in resources and this was implemented in March 2002. Cataract surgery in the region is monitored by a waiting list program that is used to track and prioritize all patients waiting. This monitored increase in funding provided an opportunity to measure exactly what impact additional resources would have on a cataract waiting list. PMID- 15540407 TI - Good environmental policy leads to good public health policy: a lesson from the grassroots. AB - Canada's environmental laws--on paper--are strong and farsighted. With proper enforcement and monitoring programs in place, no one should be at risk from contamination, no ecosystem would be threatened, and no health official should need to make such choices. Unfortunately, these laws are not always enforced, and their laudable goals not always met. PMID- 15540408 TI - Computer telephony in healthcare. AB - The telephone provides an easy-to-use, reliable and cheap computer interface. We review computer telephony applications for patient care, clinical research and medical education. We describe uses at our academic health sciences centre and discuss strengths and weaknesses of this technology. Until more sophisticated interfaces, for example personal digital assistants with wireless Internet access, are commonplace, we believe computer telephony can play a useful role in electronic healthcare. PMID- 15540409 TI - Linux thin-client conversion in a large cardiology practice: initial experience. AB - Capital Cardiology Associates (CCA) is a single-specialty cardiology practice with offices in New York and Massachusetts. In 2003, CCA converted its IT system from a Microsoft-based network to a Linux network employing Linux thin-client technology with overall positive outcomes. PMID- 15540410 TI - Focus: dictation, transcription and speech recognition 2004. PMID- 15540411 TI - How communication and co-operation eased a patient transport crisis. PMID- 15540412 TI - Use of health services and SARS. PMID- 15540413 TI - [Primary bacterial endocarditis (Part II)]. PMID- 15540414 TI - [Restenosis inside the stent: treatment and prevention]. PMID- 15540415 TI - [Use of cytokines in the complex treatment of respiratory diseases]. PMID- 15540416 TI - [Relationship of variability of blood pressure, cardiac sinus rhythm, and structural and functional characteristics of the left ventricular myocardium in elderly and senile patients with arterial hypertension]. AB - The variability of blood pressure (BP), autonomic cardiac sinus rhythm regulation, and myocardial structural and functional characteristics of the left ventricle (LV) was comparatively analyzed in 201 patients aged 65-88 years who had uncomplicated systolic and diastolic and isolated systolic arterial hypertension (SDAH and ISAH, respectively). There was a greater variability of systolic and diastolic BP no matter what the type of arterial hypertension (AH) was. It was ascertained that ISAH was characterized by absolute parasympaticotonia and SDAH was marked by relative sympaticotonia with the involvement of central ergotropic and humoral-and-metabolic mechanisms. In elderly and senile patients, uncomplicated AH was accompanied in 86.6% of cases by LV remodeling, mainly as its concentric hypertrophy (52.7%), characterizing primarily by non-restrictive diastolic dysfunction. There were no significant differences in the types of LV remodeling and diastolic dysfunction in patients with combined AH and ISAH. Moreover, the development of LV remodeling and associated diastolic dysfunction in SDAH was followed by a decrease in the overall variability of cardiac sinus rhythm with a smaller contribution of a segmental link of the parasympathetic portion of the autonomic nervous system and by the development of relative sympaticotonia due to suprasegmental and humoral and-metabolic influences. PMID- 15540417 TI - [The specific features of synthesis of heat shock proteins in patients with postinfarct cardiosclerosis]. AB - The specific features of synthesis of inducible heat shock proteins with a molecular mass of 72 kDa (HSP72i) with peripheral lymphocytes were studied in patients with postinfarct cardiosclerosis and signs of chronic heart failure (CHF). According to the baseline level and time course of changes in lymphocytic accumulation of HSP72i during treatment, the patients with CHF were divided into 3 groups. Great differences were found in the course of CHF in patients with different levels of HSP72i synthesis. There were significant relationships between the content of HSP72i and the basic clinical, laboratory, and instrumental characteristics of CHF. The baseline level of HSP72i and the time course of changes in its accumulation by peripheral lymphocytes in patients with CHF were shown to be of prognostic value. PMID- 15540418 TI - [Effect of fosinopril on the rate of neurohumoral and proinflammatory activation in patients with heart failure]. AB - The purpose of the study was to examine the effect of fosinopril on the magnitude of neurohumoral and proinflammatory activation in patients with severe heart failure (HF). Twenty-eight patients aged 52-68 years who had Functional Class (FC) III-IV HF that had developed due to coronary heart disease were examined. All the patients were divided into 2 groups (with 14 patients in each group) and they received routine therapy including an angiotensin-converting enzyme inhibitor (ACEI) and a beta-adrenoblocker. Group 1 patients were given the ACEI fosinopril in a dose of 10-20 mg/day, Group 2 patients took captopril in a dose of 50-75 mg/day. The course of therapy was 12 weeks. All the patients underwent echocardiography by the routine procedure. The plasma levels of angiotensin-II, aldosterone, and brain natriuretic peptide (BNUP) were determined by radioimmunoassay. The plasma contents of tumor necrosis factor-alpha (TNF-alpha) and C-reactive protein (C-RP) were verified by enzyme immunoassay. An analysis of the findings indicated that during therapy the FC of HF decreased on the average by 10.9% in Group 1 and by 11. 7% in Group 2 (p = 0.14). With this, fosinopril was superior to captopril not only in its capacity of improving overall left ventricular contractile function, but it was characterized by a more pronounced effect on regression of the plasma pool of C-RP and TNF-alpha. The marked depressive action of the ACEI fosinopril on the plasma pool of products of the renin-angiotensin system, BNUP, and proinflammatory cytokines may be considered to be as a basis for preferably prescribing the agent to patients with severe HF. PMID- 15540419 TI - [Gastroesophageal reflux and the degree of esophagitis in patients with coronary heart disease: impact on myocardial repolarization parameters and cardiac rhythm variability]. AB - The specific features of myocardial repolarizing processes and heart rate variability (HRV) were studied in 67 patients with coronary heart disease (CHD) concurrent with gastroesophageal reflux disease (GERD) (a study group), in 72 patients with CHD and in 46 patients with GERD (comparison groups). Patients with concomitant diseases showed a significant asynchronism of repolarizing processes and decreased HRV. A relationship was established between the presence and magnitude of inflammatory lower esophageal third changes, and the severity of myocardial repolarizing processes, and decreased HRV in patients with CHD + GERD. PMID- 15540420 TI - [Remodeling of elastic arteries in patients with hypertensive disease: diagnostic value of pulse pressure]. AB - The study was undertaken to examine a role of pulse blood pressure (PBP) as a diagnostic marker of remodeling of large elastic arteries in patients with hypertensive disease (HD). A hundred and sixty-eight patients with HD and 38 apparently healthy individuals (a control group) were examined. In all the examinees, PBP was measured and central PBP (PBPc) was calculated. According to the value of PBPc, the patients with HD were divided into 4 groups: 1) PBPc was equal or less than 45 mm Hg; 2) that was 46-50 mm Hg; 3) PBPc was 51-64 mm Hg; and 4) that was equal or more than 65 mm Hg. With Doppler study, the aortic pulse wave propagation velocity (PWPV) was determined and the aortic stiffness coefficient was (C). Seventy-seven patients with HD and 20 control individuals underwent magnetic resonance imaging with the aortic intraluminal diameter (D(ao)) and wall thickness (H(ao)) being estimated. As compared with the controls, patients with HD were found to have significantly increased PWPV, C(s), D(ao), and H(ao) which characterizes elastic arterial remodeling in HD. Comparison of these parameters in persons with different levels of PBPc demonstrated progressive dilatation, the increased stiffness and enlarged thickness of the wall of elastic arteries in those having higher PBPc. The results of a regression analysis confirmed that there is a significant relationship of PBPc with C(s), D(ao), and H(ao) and the coefficients of correlation between were 0.89, 0.71, and 0.75, respectively. The findings have indicated that the level of PBPc may serve as a marker of remodeling of large elastic arteries in patients with HD. PMID- 15540421 TI - [Effect of small-dose glucocorticoids on the course of early rheumatic arthritis]. AB - The study was undertaken to evaluate the effect of small-dose glucocorticoids (GCs) in combination with essential drugs used in early rheumatic arthritis (RA) on the clinical and laboratory activity and progression of joint destruction. Sixty-two patients aged 18-63 years who had active RA (its history being 1.5 to 24 months) and had not received basic therapy before were given methotrexate (MT) in a dose of 7.5-10.0 mg/week. Prednisolone (P) was randomly used in a dose equal or more than 10 mg/day). The efficiency of treatment was evaluated every 3 months by the ACR criteria 20/50/70. X-ray study of the hand and foot joints (the Larsen procedure by erosion calculations) was performed and the serum levels of C reactive protein (C-RP) and interleukin-6 (IL-6) were measured before and 12 months after therapy. In the MT group, the patients' mean age was 52.0 +/- 10.5 years, the history of RA was 8.4 +/- 6.8 months; 82% of the patients were seropositive in terms of rheumatoid factor; the DAS 28 index was 5.2 +/- 0.8; in the P+MT group, the above parameters were 51.9 +/- 11. 7 years, 9.1 +/- 6.0 months, 83%, and 5.4 +/- 0.8, respectively (p > 0.05). Throughout one-year follow up, the patients whose parameters corresponded to ACR 70 were more in the P+MT group than in the MT group (p < 0.05). The level of IL-6 and C-RP significantly decreased only in the P+MT group. There was a significant in the Larsen scores in both groups. A much fewer number of new erosions was revealed in the P+MT group than that in the MT group. According to the ACR 70 criteria, the efficiency of treatment with small-dose GCs was much higher than that in MT monotherapy. The small doses of GCs significantly lowered the laboratory activity of RA (C-RP, IL 6) and the occurrence of erosions. PMID- 15540423 TI - [Types of therapeutic-and-diagnostic policy for breast cystic masses]. AB - The experience gained with diagnosing and treating 228 females aged 20 to 68 years who had breast cysts has shown that its diagnosis is verified by a combination of ultrasonography, mammography, and aspiration fine-needle biopsy. The use of needle pneumoethanol therapy permits effective cure. PMID- 15540422 TI - [Effect of enalapril, lisinopril, and amlodipine on the course of chronic gastrititis in patients with arterial hypertension]. AB - A hundred and thirty-six patients with mild and moderate arterial hypertension (AH) who had received angiotensin-converting enzyme inhibitors (ACEIs) (enalapril and lisinopril) and the calcium channel blocker amlodipine were examined. The use of the above ACEIs promoted the normalization of proliferative processes in the epithelium of the gastric mucosa (GM) in patients with AH and concomitant chronic gastritis. This effect ofACEIs is not related to an antihypertensive effect since the antihypertensive agent amlodipine fails to exert such an effect on GM. The revealed fact of the positive effect ofACEIs not only on the morphological characteristics of GM, but also on the severity of the dyspeptic syndrome is important from the clinical point of view. PMID- 15540424 TI - [The prevalence of chronic generalized pain and its relationship to demographic characteristics and mental status]. AB - Chronic generalized pain (CGP) as one of the diagnostic criteria of fibromyalgia is a common condition amongst the populations of Western Europe and the USA. The prevalence of CGP in these countries is 10.7-13.2%. The etiology of CGP is unclear; however, several findings suggest that this is a process of somatization that masks affective disorders. There have been no studies of the epidemiology of CGP in Russia. The study was undertaken to determine the prevalence of CGP in the Yekaterinburg adult population and to examine the relationship of CGP to its demographic characteristics and affective disorders. A ostal cross-sectional population survey was conducted. This covered 159 persons (54 males and 105 females) aged 27 to 75 years who had been randomly selected from the election lists of Yekaterinburg residents. A questionnaire was posted to each participant in order to reveal whether a respondent had experienced pain a month before, to define its location, duration, severity, and to evaluate psychoemotional disorders. The response rate was 75.5% (120 replies). The prevalence of CGP was 13.3%. There was no relationship of the sex of a respondent to CGP. The latter was shown to be associated with age. There was also its significant association with psychoemotional disorders. CGP is a common condition in the Yekateringburg population, which occurs in every 8 adult residents, more frequently in those aged above 43 years. The individuals who suffer from CGP exhibit a high level of psychological distress. PMID- 15540425 TI - [Successful midodrin treatment for vasovagal syncopes accompanied by asystole]. AB - Vasovagal syncopes (WS) are fainting fits whose manifestations are systemic vasodilation and bradycardia. The development of WS is frequently accompanied by short-term cardiac arrest. Recent data show that implantation of a cardiac pacemaker fails to prevent the development of fainting fits in these patients. On the basis of the pathogenesis of WS, the use of alpha-adrenomimetic midodrine is justifiable. The presented case demonstrates the successful use of the agent in the treatment of vasovagal syncope accompanied by asystole. PMID- 15540426 TI - [New approaches to diagnosing and treating hyperkinetic biliary dyskinesia associated with chronic acalculous cholecystitis]. AB - Ninety patients aged 21 to 56 years who had chronic non-calculous cholecystitis (CNCC) concurrent with hyperkinetic dyskinesia (HKD) detectable by a stepwise duodenal probing and sonography, by using a choleretic breakfast and by determining the relaxation coefficient (RC) that was equal to the ratio of the volume of the gallbladder (GB) after the use of a spasmolytic to the baseline GB volume. The patients were divided into 3 groups. The authors used as a spasmolytic agent pinaverium bromide (dicetel) in a dose of 50 mg (1 tablet) in Group 1), octylonium bromide (spasmomen) in a dose of 40 mg (1 dragee) in Group 2, and drotaverine (no-spa) in a dose of 40 mg (1 tablet). There was a more significant sonographic increase in the size of GB in Groups 1 and 2 as compared with Group 3. In the acute drug test and during long-term treatment as well, the highest spasmolytic effect was noted in patients receiving dicetel (Group 1) and spasmomen (Group 2) as compared with that in Group 3 patients taking drotaverine. With this, RC was 1.25 +/- 0.2, 1.6 +/- 0.15, and 1.08 +/- 0.1, respectively. No adverse reactions occurred in the patients having selective calcium blockers (SCBs) whereas the patients receiving no-spa were found to have the following side effects: dry mouth (n = 3), transient constipation (n = 1), and numb tongue (n = 1). Thus, the study has provided evidence for the fact that SCBs have some advantage over myotropic spasmolytic agents in the treatment of CNCC with the signs of HKD. PMID- 15540427 TI - [Itch in systemic diseases]. AB - The specific features of itch in various systemic diseases are presented. A great variety of its possible mechanisms are shown in some systemic diseases. The basic principles of determination of the causes of itch and its treatment are given. PMID- 15540428 TI - [A case of inactive adrenal node that transformed to adenoma with manifestations of primary hyperaldosteronism in a patient with essential hypertension]. PMID- 15540429 TI - [Arterial hypertension: is it a disease or a risk factor?]. PMID- 15540430 TI - Highest-accuracy interferometer alignment by retroreflection scanning. AB - One important prerequisite for interferometric length measurements of high accuracy is autocollimation adjustment. This guarantees that the direction of the length scale represented by light waves is parallel to the length direction of the object investigated. First we describe the conventional visual autocollimation adjustment method used at Physikalisch-Technische Bundesanstalt since the beginning of interferometric length measurements. Then a new autocollimation method based on scanning the retroreflection from the interferometer is described. Check measurements are performed in order to investigate the quality of the adjustment. As a result of the method applied the uncertainty contribution originating from the cosine error could be reduced drastically for the interferometer used. PMID- 15540431 TI - Role of caustics in the formation of networks of amplitude zeros for partially developed speckle fields. AB - The topology of a partially developed speckle field was studied by use of interference techniques through computer simulation. Amplitude and phase structures in the vicinity of caustics for a coherent radiation field scattered at a surface with large inhomogeneities were investigated. It was confirmed that the caustics are indispensible components of the procedure for the formation of networks of amplitude zeros for a coherent field scattered by a rough surface with large inhomogeneities. It is shown that the formation of interference forklets in the field gives evidence of changes in the field's topology, as these forklets are a diagnostic sign of transition from a caustic to a three dimensional pattern of a diffraction catastrophe. PMID- 15540432 TI - Design and construction of linear laser encoders that possess high tolerance of mechanical runout. AB - A linearly diffracted laser encoder that has high tolerance of head-to-scale misalignment and a high signal-to-noise ratio is described. The preservation of parallelism between the incident and the diffracted beams, which can be attributed to a built-in folded 1x telescope, allows for the high alignment tolerance. It can be shown that, by coupling this newly developed circular polarization interferometer configuration with grating scale geometry optimization, one can eliminate the problems associated with signal distortion that arise from various efficiencies of the p- and the s-polarized light beams and obtain a high signal-to-noise ratio. Both theoretical and experimental results are presented to confirm the improved results and performance. PMID- 15540433 TI - Design and demonstration of a miniature catheter for a confocal microendoscope. AB - The fluorescence confocal microendoscope provides high-resolution, in vivo imaging of cellular pathology during optical biopsy. The confocal microendoscope employs a flexible fiber-optic catheter coupled to a custom-built slit-scan confocal microscope. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The 3-mm-diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope, adding microscopic imaging capability to conventional endoscopy. The design and performance of the miniature objective and focus assembly are discussed. Primary applications of the system include diagnosis of disease in the gastrointestinal tract and female reproductive system. PMID- 15540434 TI - Hyperosmotic chemical agent's effect on in vivo cerebral blood flow revealed by laser speckle. AB - We investigated the influence of a hyperosmotic agent (glycerol) on the normal physiological function of tissue by applying the glycerol in vitro and in vivo to rabbit dura mater to assess the changes in the tissue's optical properties. We used a laser speckle imaging technique to study the effect of epidurally applied glycerol on resting cerebral blood flow (CBF). Our results showed that resting CBF decreased as the transparency of the dura mater increased. The challenges for the design of an optical clearing technique were not only the clearing effects and the duration of the action of the chemical agents but also the influence of the glycerol on the tissue's normal physiological function. PMID- 15540435 TI - Comparison of two-color hologram lifetimes of near-stoichiometric lithium niobate and of tantalate crystals. AB - Lifetimes of two-color nonvolatile holograms recorded in undoped or in slightly doped near-stoichiometric lithium niobate and tantalate crystals were measured and compared by extrapolation of the high-temperature data. A proton-compensation mechanism dominated the dark decay and yielded similar activation energies, of 1.05 and 1.10 eV, for near-stoichiometric lithium niobate and tantalate crystals, respectively. The lifetime of holograms in lithium tantalate was 1 order of magnitude longer than that in lithium niobate with the same proton concentration, which was consistent with our theoretical estimation. The projected lifetime of two-color holograms in lithium tantalate without observable OH-absorption is longer than 50 years. PMID- 15540436 TI - N-ocular volume holographic imaging. AB - Volume holographic imaging utilizes Bragg selectivity to optically slice the object space of the imaging system and measure four- (three spatial and one spectral) dimensional object information. The N-ocular version of this method combines multiple-volume holographic sensors and digital postprocessing to yield high-resolution three-dimensional images for broadband objects located at long working distances. We discuss the physical properties of volume holography pertinent to imaging performance and describe two computational algorithms for image inversion based on filtered backprojection and least-squares optimization. PMID- 15540437 TI - Influence of thresholding on centroid statistics: full analytical description. AB - The centroid method is a common procedure for subpixel location that is applied to a large number of optical sensors. In practice, it is always accompanied by thresholding algorithms used to eliminate undesirable background that may decrease precision. We present a full analytical description of the interaction between centroiding and thresholding applied over an intensity distribution corrupted by additive Gaussian noise. An in depth analysis of the most outstanding statistical properties of this relation (mean and variance) is also presented by means of simulated and experimental data. This work provides fundamental concepts to the designers of sensors that are based on centroid measurements to allow them to use thresholding correctly before centroid computation. PMID- 15540438 TI - Integral imaging with improved depth of field by use of amplitude-modulated microlens arrays. AB - One of the main challenges in three-dimensional integral imaging is its limited depth of field. Such a limitation is imposed by diffraction, among other factors. The easiest way to improve the depth of field is by reducing the numerical aperture of the microlenses. However, such an improvement is obtained at the expense of an important deterioration in the spatial resolution. We propose a technique, which is novel in the context of integral imaging, for improving the depth of field with no deterioration of the spatial resolution. The technique, based on amplitude modulation of the array of phase elements, can substantially improve the figure of merit of the product of depth of the focus and the squared resolution. PMID- 15540439 TI - MIPAS-ENVISAT limb-sounding measurements: trade-off study for improvement of horizontal resolution. AB - The Michelson Interferometer for Passive Atmospheric Sounding (MIPAS) is a limb scanning spectrometer that has operated onboard the Environmental Satellite since the end of March 2002. Common features of limb-scanning experiments are both high vertical resolution and poor horizontal resolution. We exploit the two dimensional geo-fit retrieval approach [Appl. Opt. 40, 1872-1875 (2001)] to investigate the possibility of improving the horizontal resolution of MIPAS measurements. Two different strategies are considered for this purpose, one exploiting the possibility (offered by the geo-fit analysis method) for an arbitrary definition of the retrieval grid, the other based on the possibility of saving measurement time by degrading the spectral resolution of the interferometer. The performances of the two strategies are compared in terms of the trade-off between the attained horizontal resolution and the retrieval precision. We find that for ozone it is possible to improve by a factor of 2 the horizontal resolution, which in the nominal measurement plan is approximately 530 km. This improvement corresponds to a degradation of the retrieval precision, which on average varies from a factor of 1.4 to 2.5, depending on the adopted spectral resolution. PMID- 15540440 TI - Near-asymptotic angle dependence of ocean optical radiance. AB - The approach of ocean optical radiance to an approximate asymptotic dependence with increasing depth in spatially uniform waters is numerically examined for a variety of sea surface illumination conditions. PMID- 15540441 TI - Analysis of nonuniformities of sampled fiber Bragg gratings. AB - We have analyzed the effects of various normally distributed nonuniformities of sampled fiber Bragg gratings on the reflective spectra and group time delay. Through numerical simulations we have drawn the following conclusions: (1) the magnitude of nonuniformity with normal distribution of the fiber's average refractive-index modulation deltaneff greatly influences the characteristics of both reflective spectra and group time delay, whose suggested precision varies from 20% to 10%; (2) the nonuniformities of sampling periods P and sampling lengths L are important factors that influence the characteristics of the group time delay, and the accepted tolerance of dimensional precision of both P and L are +/- 4 microm; and (3) the tolerance of nonuniformity of the sampling period's chirp coefficient is high, and its precision can be as great as 100% with few adverse effects. PMID- 15540442 TI - Comparison of SeaWiFS measurements of the Moon with the U.S. Geological Survey lunar model. AB - The Sea-Viewing Wide-Field-of-View Sensor (SeaWiFS) has made monthly observations of the Moon since 1997. Using 66 monthly measurements, the SeaWiFS calibration team has developed a correction for the instrument's on-orbit response changes. Concurrently, a lunar irradiance model has been developed by the U.S. Geological Survey (USGS) from extensive Earth-based observations of the Moon. The lunar irradiances measured by SeaWiFS are compared with the USGS model. The comparison shows essentially identical response histories for SeaWiFS, with differences from the model of less than 0.05% per thousand days in the long-term trends. From the SeaWiFS experience we have learned that it is important to view the entire lunar image at a constant phase angle from measurement to measurement and to understand, as best as possible, the size of each lunar image. However, a constant phase angle is not required for using the USGS model. With a long-term satellite lunar data set it is possible to determine instrument changes at a quality level approximating that from the USGS lunar model. However, early in a mission, when the dependence on factors such as phase and libration cannot be adequately determined from satellite measurements alone, the USGS model is critical to an understanding of trends in instruments that use the Moon for calibration. This is the case for SeaWiFS. PMID- 15540443 TI - Highly efficient continuous-wave operation of a Nd:YAG rod laser that is side pumped by p-polarized diode laser bars. AB - We report on the performance of highly efficient, high-power continuous-wave (CW) Nd:YAG lasers that are side pumped by p-polarized diode laser beams. In this configuration pump light is directly coupled into the Nd:YAG rod through a threefold symmetric gold-coated flow tube. The polarization direction of our pump diode bars is perpendicular to the rod's axis (p-polarized). In a closed coupled resonator, a maximum output power of 195 W in multimode operation is obtained for a pump power of 423 W, which corresponds to an optical-to-optical efficiency of 46% and an electrical-to-optical efficiency of 23%. This is, to the best of our knowledge, the highest electrical-to-optical efficiency reported for a CW diode side-pumped Nd:YAG laser. By the pump-power leakage analysis method, we measured the pumping efficiency to be approximately 94%. The high efficiency of the system can be attributed to wing pumping, which results in uniform pump-light distribution and better pumping efficiency because of the use of p-polarized pump beams. PMID- 15540444 TI - Diode-laser frequency stabilization by two-frequency Doppler-broadened absorption spectroscopy. AB - We demonstrate a robust method for frequency stabilization of a diode laser by two Doppler-broadened absorption spectra of the cesium D2 line. This technique employs an acousto-optical modulator to generate another frequency component from a diode laser to perform the spectroscopy. The 852-nm diode laser with frequency stabilization at the zero crossing of the error signal showed a peak-to-peak fluctuation of 800 kHz compared with a frequency-stabilized femtosecond laser over a 2-h period. This frequency-locking method is free of modulation and can be applied to frequency discriminators as well as to atomic resonances. PMID- 15540445 TI - Multireflection pumping concept for miniaturized diode-pumped solid-state lasers. AB - An innovative pump concept for diode-pumped, solid-state lasers is introduced as an example for an Er:YSGG laser, permitting its miniaturization. Embedded in a multireflective pump cavity, the laser crystal is simultaneously side and end pumped. Specially calculated and shaped deflecting optics distribute the coaxially input pumping light homogeneously over the lateral surface of the crystal, therefore reducing the size of the laser head, including the optical resonator, to a length of 27.5 mm and an outside diameter of 12.5 mm. The differential efficiency achieved is between 8.7% and 24%. The laser emits energy of 15.7 mJ at an absolute efficiency of 9.1% and a repetition rate of 4 Hz. PMID- 15540446 TI - Passive standoff detection of chemical warfare agents on surfaces. AB - Results are presented on the passive standoff detection and identification of chemical warfare (CW) liquid agents on surfaces by the Fourier-transform IR radiometry. This study was performed during surface contamination trials at Defence Research and Development Canada-Suffield in September 2002. The goal was to verify that passive long-wave IR spectrometric sensors can potentially remotely detect surfaces contaminated with CW agents. The passive sensor, the Compact Atmospheric Sounding Interferometer, was used in the trial to obtain laboratory and field measurements of CW liquid agents, HD and VX. The agents were applied to high-reflectivity surfaces of aluminum, low-reflectivity surfaces of Mylar, and several other materials including an armored personnel carrier. The field measurements were obtained at a standoff distance of 60 m from the target surfaces. Results indicate that liquid contaminant agents deposited on high reflectivity surfaces can be detected, identified, and possibly quantified with passive sensors. For low-reflectivity surfaces the presence of the contaminants can usually be detected; however, their identification based on simple correlations with the absorption spectrum of the pure contaminant is not possible. PMID- 15540447 TI - Chlorophyll biomass in the global oceans: satellite retrieval using inherent optical properties. AB - In the upper layer of the global ocean, 2082 in situ chlorophyll biomass values (Chl) are retrieved by concurrent satellite-derived inherent optical properties (IOP). It is found that (1) the phytoplankton absorption coefficient IOP alone does not provide satisfactory (Chl) retrieval; (2) the chromophoric dissolved organic matter (CDOM) absorption coefficient IOP must also be used to obtain satisfactory retrieval through (Chl) alpha a ph + pa CDOM where p is a constant and a ph and aCDOM are, respectively, the phytoplankton and CDOM absorption coefficients; (3) the IOP-based (Chl) retrieval performance is comparable to standard satellite reflectance ratio retrievals (that have CDOM absorption intrinsically embedded within them); (4) inclusion of the total backscattering coefficient IOP does not contribute significantly to (Chl) retrieval; and (5) the new IOP-based algorithm may provide the possibility for future research to establish the actual role of extracellular CDOM from all sources in the intracellular production of chlorophyll biomass. PMID- 15540448 TI - Modeling of the signals of an optical particle counter for real nonspherical particles. AB - An optical particle counter (OPC) was exposed to atmospheric particles of diameters of 200, 300, and 400 nm. The OPC data were combined with the results of single-particle analysis with a transmission electron microscope (TEM) on samples taken in parallel with the OPC measurements. With a T-matrix-based optical model the measured OPC spectra of scattered light pulses could be approximated with good precision. With an algorithm that simulated the response of the OPC to a given population of model particles derived from the TEM results, average absorption properties of different particle types were retrieved. For mobility sizes of 400 nm, higher light absorption was retrieved with the optical model for soot aggregates than for the rest of the morphological particle types. At smaller mobility sizes no compositional information could be derived from the model particles derived from the TEM data. Despite the limited success of the new methodology applied to the present experiment the results encourage the use of OPCs in combination with electrical mobility analyzers to derive more than aerosol-size distributions. With state-of-the-art pulse-height analysis the light scattering pulses could be resolved with much finer resolution than in the instrument used. PMID- 15540449 TI - Rayleigh-calibrated fluorescence quantum yield measurements of acetone and 3 pentanone. AB - We measured fluorescence quantum yields of acetone and 3-pentanone as a pure gas and with nitrogen diluent at room temperature at 20, 507, and 1013 mbar using 248, 266, and 308 nm excitation by calibrating the optical collection system with Rayleigh scattering from nitrogen. At 20 mbar with 308-nm excitation, the fluorescence quantum yields for acetone and 3-pentanone are 7 +/- 1 x 10(-4) and 1.1 +/- 0.2 x 10(-3), respectively, and each decreases with decreasing excitation wavelength. These directly measured values are significantly lower than earlier ones that were based on a chain of relative measurements. The observed pressure and excitation wavelength dependence is in qualitative agreement with a previously developed fluorescence quantum yield model, but the absolute numbers disagree. Changing acetone's fluorescence rate constant to 3 x 10(5) s(-1) from its previous value of 8 x 10(5) s(-1) resulted in good agreement between our measurements and the model. PMID- 15540450 TI - A survey on palette reordering methods for improving the compression of color indexed images. AB - Palette reordering is a well-known and very effective approach for improving the compression of color-indexed images. In this paper, we provide a survey of palette reordering methods, and we give experimental results comparing the ability of seven of them in improving the compression efficiency of JPEG-LS and lossless JPEG 2000. We concluded that the pairwise merging heuristic proposed by Memon et al. is the most effective, but also the most computationally demanding. Moreover, we found that the second most effective method is a modified version of Zeng's reordering technique, which was 3%-5% worse than pairwise merging, but much faster. PMID- 15540451 TI - An efficient re-indexing algorithm for color-mapped images. AB - The efficiency of lossless compression algorithms for fixed-palette images (indexed images) may change if a different indexing scheme is adopted. Many lossless compression algorithms adopt a differential-predictive approach. Hence, if the spatial distribution of the indexes over the image is smooth, greater compression ratios may be obtained. Because of this, finding an indexing scheme that realizes such a smooth distribution is a relevant issue. Obtaining an optimal re-indexing scheme is suspected to be a hard problem and only approximate solutions have been provided in literature. In this paper, we restate the re indexing problem as a graph optimization problem: an optimal re-indexing corresponds to the heaviest Hamiltonian path in a weighted graph. It follows that any algorithm which finds a good approximate solution to this graph-theoretical problem also provides a good re-indexing. We propose a simple and easy-to implement approximation algorithm to find such a path. The proposed technique compares favorably with most of the algorithms proposed in literature, both in terms of computational complexity and of compression ratio. PMID- 15540452 TI - Improved structures of maximally decimated directional filter banks for spatial image analysis. AB - This paper introduces an improved structure for directional filter banks (DFBs) that preserves the visual information in the subband domain. The new structure achieves this outcome while preserving both the efficient polyphase implementation and the exact reconstruction property. The paper outlines a step by-step framework in which to examine the DFB, and within this framework discusses how, through the insertion of post-sampling matrices, visual distortions can be removed. In addition to the efficient tree structure, attention is given to the form and design of efficient linear phase filters. Most notably, linear phase IIR prototype filters are presented, together with the design details. These filters can enable the DFB to have more than a three-fold improvement in complexity reduction over quadrature mirror filters (QMFs). PMID- 15540453 TI - Dense motion estimation using regularization constraints on local parametric models. AB - This paper presents a method for dense optical flow estimation in which the motion field within patches that result from an initial intensity segmentation is parametrized with models of different order. We propose a novel formulation which introduces regularization constraints between the model parameters of neighboring patches. In this way, we provide the additional constraints for very small patches and for patches whose intensity variation cannot sufficiently constrain the estimation of their motion parameters. In order to preserve motion discontinuities, we use robust functions as a regularization mean. We adopt a three-frame approach and control the balance between the backward and forward constraints by a real-valued direction field on which regularization constraints are applied. An iterative deterministic relaxation method is employed in order to solve the corresponding optimization problem. Experimental results show that the proposed method deals successfully with motions large in magnitude, motion discontinuities, and produces accurate piecewise-smooth motion fields. PMID- 15540454 TI - A regularized curvature flow designed for a selective shape restoration. AB - Among all filtering techniques, those based exclusively on image level sets (geometric flows) have proven to be the less sensitive to the nature of noise and the most contrast preserving. A common feature to existent curvature flows is that they penalize high curvature, regardless of the curve regularity. This constitutes a major drawback since curvature extreme values are standard descriptors of the contour geometry. We argue that an operator designed with shape recovery purposes should include a term penalizing irregularity in the curvature rather than its magnitude. To this purpose, we present a novel geometric flow that includes a function that measures the degree of local irregularity present in the curve. A main advantage is that it achieves non trivial steady states representing a smooth model of level curves in a noisy image. Performance of our approach is compared to classical filtering techniques in terms of quality in the restored image/shape and asymptotic behavior. We empirically prove that our approach is the technique that achieves the best compromise between image quality and evolution stabilization. PMID- 15540455 TI - Statistical modeling of complex backgrounds for foreground object detection. AB - This paper addresses the problem of background modeling for foreground object detection in complex environments. A Bayesian framework that incorporates spectral, spatial, and temporal features to characterize the background appearance is proposed. Under this framework, the background is represented by the most significant and frequent features, i.e., the principal features, at each pixel. A Bayes decision rule is derived for background and foreground classification based on the statistics of principal features. Principal feature representation for both the static and dynamic background pixels is investigated. A novel learning method is proposed to adapt to both gradual and sudden "once off" background changes. The convergence of the learning process is analyzed and a formula to select a proper learning rate is derived. Under the proposed framework, a novel algorithm for detecting foreground objects from complex environments is then established. It consists of change detection, change classification, foreground segmentation, and background maintenance. Experiments were conducted on image sequences containing targets of interest in a variety of environments, e.g., offices, public buildings, subway stations, campuses, parking lots, airports, and sidewalks. Good results of foreground detection were obtained. Quantitative evaluation and comparison with the existing method show that the proposed method provides much improved results. PMID- 15540456 TI - Spatiotemporal motion boundary detection and motion boundary velocity estimation for tracking moving objects with a moving camera: a level sets PDEs approach with concurrent camera motion compensation. AB - The purpose of this study is to investigate a method of tracking moving objects with a moving camera. This method estimates simultaneously the motion induced by camera movement. The problem is formulated as a Bayesian motion-based partitioning problem in the spatiotemporal domain of the image quence. An energy functional is derived from the Bayesian formulation. The Euler-Lagrange descent equations determine imultaneously an estimate of the image motion field induced by camera motion and an estimate of the spatiotemporal motion undary surface. The Euler-Lagrange equation corresponding to the surface is expressed as a level-set partial differential equation for topology independence and numerically stable implementation. The method can be initialized simply and can track multiple objects with nonsimultaneous motions. Velocities on motion boundaries can be estimated from geometrical properties of the motion boundary. Several examples of experimental verification are given using synthetic and real-image sequences. PMID- 15540457 TI - Visual tracking and recognition using appearance-adaptive models in particle filters. AB - We present an approach that incorporates appearance-adaptive models in a particle filter to realize robust visual tracking and recognition algorithms. Tracking needs modeling interframe motion and appearance changes, whereas recognition needs modeling appearance changes between frames and gallery images. In conventional tracking algorithms, the appearance model is either fixed or rapidly changing, and the motion model is simply a random walk with fixed noise variance. Also, the number of particles is typically fixed. All these factors make the visual tracker unstable. To stabilize the tracker, we propose the following modifications: an observation model arising from an adaptive appearance model, an adaptive velocity motion model with adaptive noise variance, and an adaptive number of particles. The adaptive-velocity model is derived using a first-order linear predictor based on the appearance difference between the incoming observation and the previous particle configuration. Occlusion analysis is implemented using robust statistics. Experimental results on tracking visual objects in long outdoor and indoor video sequences demonstrate the effectiveness and robustness of our tracking algorithm. We then perform simultaneous tracking and recognition by embedding them in a particle filter. For recognition purposes, we model the appearance changes between frames and gallery images by constructing the intra- and extrapersonal spaces. Accurate recognition is achieved when confronted by pose and view variations. PMID- 15540458 TI - Polygonal and polyhedral contour reconstruction in computed tomography. AB - This paper is about three-dimensional (3-D) reconstruction of a binary image from its X-ray tomographic data. We study the special case of a compact uniform polyhedron totally included in a uniform background and directly perform the polyhedral surface estimation. We formulate this problem as a nonlinear inverse problem using the Bayesian framework. Vertice estimation is done without using a voxel approximation of the 3-D image. It is based on the construction and optimization of a regularized criterion that accounts for surface smoothness. We investigate original deterministic local algorithms, based on the exact computation of the line projections, their update, and their derivatives with respect to the vertice coordinates. Results are first derived in the two dimensional (2-D) case, which consists of reconstructing a 2-D object of deformable polygonal contour from its tomographic data. Then, we investigate the 3-D extension that requires technical adaptations. Simulation results illustrate the performance of polygonal and polyhedral reconstruction algorithms in terms of quality and computation time. PMID- 15540459 TI - A multiplicative regularization approach for deblurring problems. AB - In this work, an iterative inversion algorithm for deblurring and deconvolution is considered. The algorithm is based on the conjugate gradient scheme and uses the so-called weighted L2-norm regularizer to obtain a reliable solution. The regularizer is included as a multiplicative constraint. In this way, the appropriate regularization parameter will be controlled by the optimization process itself. In fact, the misfit in the error in the space of the blurring operator is the regularization parameter. Then, no a priori knowledge on the blurred data or image is needed. If noise is present, the misfit in the error consisting of the blurring operator will remain at a large value during the optimization process; therefore, the weight of the regularization factor will be more significant. Hence, the noise will, at all times, be suppressed in the reconstruction process. Although one may argue that, by including the regularization factor as a multiplicative constraint, the linearity of the problem has been lost, careful analysis shows that, under certain restrictions, no new local minima are introduced. Numerical testing shows that the proposed algorithm works effectively and efficiently in various practical applications. PMID- 15540460 TI - Unsupervised learning of a finite mixture model based on the Dirichlet distribution and its application. AB - This paper presents an unsupervised algorithm for learning a finite mixture model from multivariate data. This mixture model is based on the Dirichlet distribution, which offers high flexibility for modeling data. The proposed approach for estimating the parameters of a Dirichlet mixture is based on the maximum likelihood (ML) and Fisher scoring methods. Experimental results are presented for the following applications: estimation of artificial histograms, summarization of image databases for efficient retrieval, and human skin color modeling and its application to skin detection in multimedia databases. PMID- 15540461 TI - Neuroligin 2 is exclusively localized to inhibitory synapses. AB - Neuroligins are cell adhesion proteins that are thought to instruct the formation and alignment of synaptic specializations. The three known rodent neuroligin isoforms share homologous extracellular acetylcholinesterase-like domains that bridge the synaptic cleft and bind beta-neurexins. All neuroligins have identical intracellular C-terminal motifs that bind to PDZ domains of various target proteins. Neuroligin 1 is specifically localized to glutamatergic postsynaptic specializations. We show here that neuroligin 2 is exclusively localized to inhibitory synapses in rat brain and dissociated neurons. In immature neurons, neuroligin 2 is found at synapses and also at GABAA receptor aggregates that are not facing presynaptic termini, indicating that postsynaptic mechanisms lead to synaptic recruitment of neuroligin 2. Our findings identify neuroligin 2 as a new cell adhesion protein specific for inhibitory synapses and open new avenues for identifiying the constituents of this unique type of postsynaptic specialization. PMID- 15540462 TI - Most genes encoding cytoplasmic intermediate filament (IF) proteins of the nematode Caenorhabditis elegans are required in late embryogenesis. AB - Intestinal cells of C. elegans show an unexpectedly high complexity of cytoplasmic intermediate filament (IF) proteins. Of the 11 known IF genes six are coexpressed in the intestine, i.e. genes B2, C1, C2, D1, D2, and E1. Specific antibodies and GFP-promoter constructs show that genes B2, D1, D2, and E1 are exclusively expressed in intestinal cells. Using RNA interference (RNAi) by microinjection at 25 degrees C rather than at 20 degrees C we observe for the first time lethal phenotypes for C1 and D2. RNAi at 25 degrees C also shows that the known A1 phenotype occurs already in the late embryo after microinjection and is also observed by feeding which was not the case at 20 degrees C. Thus, RNAi at 25 degrees C may also be useful for the future analysis of other nematode genes. Finally, we show that triple RNAi at 20 degrees C is necessary for the combinations B2, D1, E1 and B2, D1, D2 to obtain a phenotype. Together with earlier results on genes A1, A2, A3, B1, and C2 RNAi phenotypes are now established for all 11IF genes except for the A4 gene. RNAi phenotypes except for A2 (early larval lethality) and C2 (adult phenotype) relate to the late embryo. We conclude that in C. elegans cytoplasmic IFs are required for tissue integrity including late embryonic stages. This is in strong contrast to the mouse, where ablation of IF genes apparently does not affect the embryo proper. PMID- 15540463 TI - Interferon-gamma acts proapoptotic on hepatic stellate cells (HSC) and abrogates the antiapoptotic effect of interferon-alpha by an HSP70-dependant pathway. AB - The activated hepatic stellate cell (HSC) is an important fibrogenic cell type of the liver. Interferon-alpha (IFN-alpha) has recently been shown to elicit an antiapoptotic effect on activated HSC by a JAK-2-dependent inhibition of caspase 8 activation. As JAK-2 has so far been shown to be a member of the IFN-gamma signal transduction pathway we studied the effect of IFN-gamma on apoptosis as well as on its signaling in primary cultured rat HSC. IFN-gamma elicited a proapoptotic effect in activated HSC. The combination of both, IFN-gamma and IFN alpha, however, completely cancelled each other's effect. No effect of the two cytokines on major members of apoptosis-regulating systems (CD95, CD95L, bcl-2, bax, bcl-xL, p53, p21WAF1, p27, NFkappaB) could be observed. Western Blot analysis revealed that gene expression of the chaperone HSP70 was found to be downregulated by IFN-gamma but upregulated by IFN-alpha. The effect could be abrogated by administration of both. After transfection of activated HSC with a pCMV-HSP70 M expression vector the proapoptotic effect of IFN-gamma was cancelled. Using HSP70 antisense, the antiapoptotic effect of IFN-alpha was cancelled as well. However IFN-gamma had no effect on upregulation of JAK-2 and pJAK-2 by IFN-alpha. Taken together IFN-gamma and IFN-alpha exert opposite effects on apoptosis in HSC. This effect is mediated by their counteracting effect on HSP70 expression which acts antiapoptotic at the level of caspase-8. PMID- 15540464 TI - Caspase-3-like protease influences but is not essential for DNA fragmentation in Blastocystis undergoing apoptosis. AB - Blastocystis hominis undergo apoptosis after treatment with a cytotoxic monoclonal antibody (MAb), 1D5, by mechanisms that are not fully understood, although our previous study demonstrated that caspase-3-like protease activity is involved. To elucidate the mechanism of MAb 1D5-induced apoptosis, we inhibited Blastocystis caspase-3-like protease to investigate if there would be a concomitant decrease in in situ DNA fragmentation. However, MAb 1D5-induced apoptosis, evidenced by DNA fragmentation, was not completely blocked by pretreating with specific caspase-3 inhibitor, Ac-DEVD-CHO, indicating that caspase-independent apoptotic pathways might also be involved. Our results also revealed that the treatment with MAb 1D5 resulted in the loss of mitochondrial membrane potential (deltapsim), independent of Ac-DEVD-CHO pretreatment. In conclusion, this study demonstrates that MAb 1D5-induced apoptosis in B. hominis is not wholly dependent on caspase-3-like protease activity and is associated with mitochondrial dysregulation. This is the first report showing evidence for complex apoptotic pathways in a unicellular parasite. PMID- 15540465 TI - Genetic analysis of fibrosarcoma of bone, a rare tumour entity closely related to osteosarcoma and malignant fibrous histiocytoma of bone. AB - Fibrosarcoma (FS) of bone is an extremely rare and genetically uncharacterised malignant tumour arising in the skeleton. On the basis of clinicopathologic features it appears to be closely related to either fibroblastic osteosarcoma (OS) or malignant fibrous histiocytoma (MFH) of bone. In this study, 27 decalcified, paraffin-embedded FS of bone were collected for genetic and immunohistochemical characterisation. Good quality DNA, suitable for genetic analyses, was isolated from nine cases (7 primary tumours, 1 local recurrence, and 1 lung metastasis), which were analysed by comparative genomic hybridisation (CGH) on chromosomes and DNA microarrays. DNA sequence copy number changes were found in five out of seven primary tumours (72%), as well as in both, the local recurrence and the metastatic lesion, by CGH on chromosomes. The most frequent aberration was gain of the chromosomal region 22q, which was present in four out of the five primary tumours with genetic changes, in the local recurrence and, as the sole genetic aberration, in the lung metastasis. DNA microarray analysis showed that gain of the platelet-derived growth factor beta (PDGF-B) gene (located at 22q12.3-q13.1) was the most frequent gene imbalance, which was present in three out of the five analysed tumours. In these three cases, real time PCR revealed a 2.1- to 2.7-fold increase of PDGF-B gene copy numbers. By immunohistochemistry, a positive reaction for B-chain-containing PDGF proteins was revealed in all the cases showing gain of 22q. A more extensive immunohistochemical analysis identified the presence of PDGF-B proteins in 8/20 primary FS of bone (40%), 3/3 lung metastases and in 1/2 local recurrences. A simultaneous positive reaction for PDGF-B proteins and PDGF receptors was found in two third of PDGF-B-positive cases (8/12). Taken together, the genetic and immunohistochemical data indicate that over-representation of the chromosomal region 22q, including particularly the PDGF-B gene, may be important for the pathogenesis of FS of bone. Our results also demonstrate that CGH on chromosomes and DNA microarrays are suitable for the genetic characterisation of decalcified, paraffin-embedded tumour tissue samples and may facilitate, combined with other techniques, the rapid acquisition of data providing insight into the molecular genetic and biologic basis of rare bone sarcomas. Moreover, these findings suggest the possible presence of an autocrine loop in FS of bone, which might be taken into account for planning innovative therapeutic strategies for patients unresponsive to conventional treatments. PMID- 15540466 TI - Lipid transfer proteins in Parietaria judaica L. pollen grains: immunocytochemical localization and function. AB - Parietariajudaica L. (Urticaceae) pollen is considered one of the most common causes of allergic respiratory symptoms in the Mediterranean area. The localization of lipid transfer proteins (LTPs) in P. judaica mature and hydrated activated pollen grains was investigated applying a combination of transmission electron microscopy (TEM) with immunocytochemical methods. Our results show that the content of LTPs in P. judaica pollen grains changes during the process of hydration. The localization of judaica LTPs in the cytoplasm and in the lipid bodies associated with vacuoles demonstrated that LTPs represent primarily intracellular proteins. On the other hand, exposure of the pollen grains to germination medium induced the release of LTPs from the pollen grain. Thus, LTPs are cytoplasmic proteins that are secreted to become available for pollen-stigma interactions and probably induce the IgE antibody responses in allergic patients. PMID- 15540467 TI - Calculate cost per RVU to establish capitation strategy. PMID- 15540468 TI - Cap shows resilience in physician compensation, production benchmarks. PMID- 15540470 TI - UR, other cost containment measures reappear in managed care strategies. PMID- 15540469 TI - Top health care spending categories spell opportunity for capitated docs. PMID- 15540471 TI - Medicare holds lottery for access to expensive drugs. PMID- 15540472 TI - New antimicrobial advisory released for prophylaxis in surgery. PMID- 15540473 TI - Conference tackles overprescribing of antimicrobials. PMID- 15540474 TI - 'Locked storage' addressed in CMS's new interpretive guidelines. PMID- 15540475 TI - CMS panel examines states' challenges with Medicare reform. PMID- 15540476 TI - Dietary supplements: first do some harm. PMID- 15540477 TI - Acute postoperative hypertension: a review of therapeutic options. AB - PURPOSE: The pathophysiology and treatment of acute postoperative hypertension (APH) are discussed. SUMMARY: APH is a significant elevation in arterial blood pressure (BP) during the immediate postoperative period. The predominant underlying mechanism appears to be sympathetic activation. APH may lead to serious neurologic, cardiovascular, or surgical-site complications and often requires intervention and management. Postoperative hypertension lasts less than six hours in most patients. Reversible or treatable causes of hypertension, including pain, anxiety, hypothermia, and hypoxemia, should be considered and treated before the implementation of antihypertensive therapy. The ideal agent for treating APH is intravenously administered, is fast acting, and has a short duration of action, allowing the rapid and safe adjustment of therapy to achieve a targeted BP range. Sodium nitroprusside has long been considered the standard therapy and has many of the ideal characteristics. However, because of the need for invasive hemodynamic monitoring and concerns about toxicity in patients given sodium nitroprusside, several newer agents may be preferable in routine clinical practice. Labetalol, nicardipine, and nitroglycerin have been widely studied or used. Hydralazine, esmolol, fenoldopam, angiotensin-converting-enzyme inhibitors, and clonidine may also be useful treatment options. CONCLUSION: When treatment of APH is necessary, therapy should be individualized for the patient. No one agent is preferred, but effective options include sodium nitroprusside, nitroglycerin, labetalol, and nicardipine. PMID- 15540478 TI - HMG-CoA reductase inhibitors for lowering elevated levels of C-reactive protein. AB - PURPOSE: Clinical trials evaluating the effectiveness of therapy with hydroxymethylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors for reducing elevated C-reactive protein (CRP) levels and associated coronary events are reviewed. SUMMARY: Atherosclerotic plaque growth may be attenuated with therapy aimed at minimizing inflammation. Because increased levels of CRP have been associated with arterial-wall inflammation, statins can prevent ischemia by both inhibiting deposition of lipids and decreasing inflammation. Evaluation of recent clinical trials, including WOSCOPS, PRINCE, AFCAPS/ TexCAPS, MIRACL, CURVES, REVERSAL, and JUPITER, demonstrated the correlation of statin therapy with decreased levels of CRP. WOSCOPS found that patients with CRP values of > 4.59 mg/L at baseline were at the highest risk of coronary events. The PRINCE trial evaluated the antiinflammatory effects of pravastatin and found a mean 16.9% reduction in CRP levels after 24 weeks of therapy. AFCAPS/TexCAPS researchers found that lovastatin provded a 14.8% reduction in the median levels of CRP (p < 0.001). The MIRACL study showed that atorvastatin reduced CRP levels by 83% (p < 0.001). Researchers in the CURVES study found a significant reduction in CRP levels with pravastatin, simvastatin, and atorvastatin compared with baseline (p < 0.025). Results of the REVERSAL study linked atorvastatin with a 36.4% decrease in CRP levels, while pravastatin was associated with a 5.2% decrease (p < 0.0001). JUPITER is ongoing and will determine whether long-term use of rosuvastatin can reduce the rate of coronary events. CONCLUSION: The lowering of elevated CRP levels by statins may reduce the risk of coronary events independently of the effect of statins on lipid levels. PMID- 15540479 TI - Stability of meropenem in polyvinyl chloride bags and an elastomeric infusion device. AB - PURPOSE: The stability of meropenem in i.v. solutions stored in polyvinyl chloride (PVC) bags and an elastomeric infusion device at concentrations commonly used in home care was studied. METHODS: Vials of meropenem were reconstituted with sterile water for injection and mixed with 0.9% sodium chloride injection (NS) to yield concentrations of 4, 10, and 20 mg/mL. Six replicate solutions were prepared in PVC containers and six in the Homepump ECLIPSE elastomeric infusion device. All solutions were stored at an average temperature of 5 degrees C and sampled immediately after preparation and at intervals up to 120 hours (five days); the 4-mg/mL solution was also sampled at 144 and 168 hours (seven days). Samples were assayed for meropenem concentration by stability-indicating high performance liquid chromatography. RESULTS: All solutions of meropenem retained over 90% of the initial drug concentration at five days. The 4-mg/mL solutions retained over 93% of the initial concentration at seven days. The rate of meropenem decay did not differ significantly between PVC and elastomeric infusion containers for the 4- and 20-mg/mL solutions; however, there was a difference for the 10-mg/mL solutions. CONCLUSION: Meropenem 4 mg/mL in NS was stable for at least seven days in PVC bags and elastomeric infusion containers when stored at 5 degrees C, and meropenem 10 and 20 mg/mL in NS was stable for at least five days in both containers at 5 degrees C. PMID- 15540480 TI - Understanding and preparing for pharmacy practice residency interviews. PMID- 15540481 TI - Reconciliation of discrepancies in medication histories and admission orders of newly hospitalized patients. PMID- 15540482 TI - Program to restrict use of i.v. fluconazole. PMID- 15540483 TI - Responding to requests for dietary supplements. PMID- 15540484 TI - Outcomes of an antimicrobial control program in a Dutch hospital. PMID- 15540485 TI - ASHP statement on the use of dietary supplements. PMID- 15540486 TI - Intranasal vaccine for prevention of influenza in children. AB - The current strategy for combating the yearly toll of influenza is to protect the highest-risk individuals through annual immunization. Despite its effectiveness, the inactivated influenza vaccine is underused, and no substantial decrease in influenza-related morbidity and mortality has occurred during the past two decades. Increasing recognition of (1) the high rate of influenza and the substantial morbidity it causes in otherwise healthy children and (2) the dominant role of children in community transmission of the influenza virus has led to the proposal for routine immunization of healthy children. This strategy would both reduce the medical burden of influenza in children and limit the spread of the disease. FluMist, because of its ease of administration, tolerability, and safety, is well suited for mass immunization. The success of such an immunization policy will, however, be predicated on addressing organizational and logistical challenges. Pharmacists will be key in developing the educational foundation and organizational framework for such expanded coverage. PMID- 15540487 TI - Green rockets. PMID- 15540488 TI - [Brazilian contribution to universal Cardiology. Subsidy to the SBC in its 60 year of activity]. PMID- 15540489 TI - Correlates of drug treatment outcomes for African American and white male federal prisoners: results from the TRIAD study. AB - The purpose of this study was to compare the effects of family background and preincarceration socioenvironmental variables on three-year post-release drug use for African American and white prison-based drug treatment participants in order to explain the previously found disparities in rates of three-year post-release drug use between the two groups. There were two hypotheses: 1) for both groups, family background and preincarceration socioenvironmental variables would predict postrelease drug use more strongly than sociodemographic characteristics and preincarceration behaviors, and 2) the predictors would be different for each group. The sample included 279 African American and 512 white male treatment participants who were supervised by a U.S. probation officer following incarceration. Event history analyses were used to model time to first drug use during postrelease supervision. The results indicated that none of the family background factors or socioenvironmental variables predicted postrelease drug use. The variables predictive of drug use for one or both racial groups were limited to sociodemographic characteristics and preincarceration behaviors such as age at release, prior commitments, and preincarceration employment. Yet, there were no significant between-group differences for these predictors. The authors concluded that future assessment of the effects of socioenvironmental variables on postrelease drug use likely requires evaluation of the postrelease social environment at the time of release. PMID- 15540490 TI - A psychometric study of the prevalence of DSM-IV personality disorders among office-based methadone maintenance patients. AB - Using the DSM-IV criteria for personality disorders, prevalence rates for these disorders were evaluated among methadone maintenance patients, with a psychometric test--the Millon Clinical Multiaxial Inventory (MCMI-III). We found that 77% of patients met the study criteria for at least one personality disorder. Of those who had a personality disorder, 20% had two personality disorders, 14% had three personality disorders, and 6% had four personality disorders. Rates of specific personality disorders are reported. Consistencies and divergence from existing research literature are noted. It is suggested that future research compare psychometrically based self-report questionnaires to a structured clinical interview format, within the same clinical population. PMID- 15540491 TI - Which substance abuse treatment facilities offer dual diagnosis programs? AB - This report examines the distribution of specialized programs or groups for dual diagnosis clients in substance abuse treatment facilities across the United States and the availability of services often needed by this client population at these facilities. Data from the National Survey of Substance Abuse Treatment Services (N-SSATS) were used to assess the geographic distribution and the characteristics of facilities that offer dual diagnosis treatment programs/groups and to examine the other clinical, social, and health services available in these facilities. Overall, 49.9% of all U.S. substance abuse treatment facilities offered dual diagnosis programs/groups. There were no clear regional variations on availability of these services. There were, however, clear variations based on facility ownership and treatment modality. Facilities owned by the federal, state, and local governments, and those offering inpatient treatment were more likely than were other facilities to offer dual diagnosis programs/groups. A large proportion of facilities with specialized dual diagnosis programs/ groups did not offer mental, social, or health services often needed by dual diagnosis clients. It is concluded that specialized programs for dual diagnosis clients are offered in many different substance abuse treatment facilities, attesting to the growing recognition of the special needs of this group of clients. However, dissemination of comprehensive services often needed to meet the special needs of these clients has not kept pace with the spread of specialized programs, highlighting the need for establishing and implementing standard evidence-based guidelines for dual diagnosis treatment programs in these settings. PMID- 15540492 TI - Effectiveness of long-term residential substance abuse treatment for women: findings from three national studies. AB - The effectiveness of residential substance abuse treatment for women was examined using data from the Center for Substance Abuse Treatment's Residential Women and Children/Pregnant and Postpartum Women (RWC/PPW) Cross-Site Study and two other recent national studies. Treatment success was defined as posttreatment abstinence from further drug or alcohol use, measured through in-person follow-up interviews conducted 6-12 months after each client's discharge. Despite differences in treatment programs, client profiles, follow-up intervals, data collection methods, and other factors, all three studies found high treatment success rates--ranging narrowly from 68% to 71% abstinent--among women who spent six months or more in treatment. Success rates were lower, and between-study differences were larger, for clients with shorter stays in treatment. Controlling for salient client and treatment project characteristics, strong associations between length of stay in treatment and posttreatment abstinence rate were found in all three studies, suggesting that women's length of stay in residential treatment is a major determinant of treatment effectiveness. In further analysis of RWC/PPW data, treatment completion was also found to be an important outcome factor. Among clients who remained in treatment for at least three months, those who achieved their treatment goals in three to five months abstinence outcomes were as good as those for clients who took more than six months to complete their treatment (76%-78% abstinent) and substantially better than those for clients who did not complete treatment (51%-52% abstinent). Notably, however, most of the RWC/PPW clients who successfully completed treatment (71%) required six months or more to do so. PMID- 15540494 TI - Psychiatric severity and spirituality, helping, and participation in alcoholics anonymous during recovery. AB - Although helping others is a critical part of Alcoholics Anonymous (AA) and many treatment programs, measures for assessing helping and describing its relationship with sobriety are lacking. A sample of 200 subjects completed a Helper Therapy Scale including three subscales: Recovery Helping (alpha = 0.78), Life Helping (alpha = 0.62), and Community Helping (alpha = 0.60). A previous analysis using structural equation modeling found that length of sobriety predicted measures of spirituality, helping, and AA participation. The analysis reported here examined whether psychiatric severity was associated with these variables. Results indicated significant relationships between psychiatric severity and measures of spirituality (Self Transcendence, Forgiveness, Positive Coping, and Negative Coping) and AA Achievement (defined as completing the 12 steps and serving as a sponsor). However, no relationships were found between psychiatric severity and length of sobriety, the three Helper Therapy subscales, or AA involvement. The findings suggest that individuals with higher psychiatric severity may need assistance from their peers or professional service providers to develop a spiritual life, serve as a sponsor for others, or complete the steps of AA. PMID- 15540493 TI - Drug seeking behavior. AB - CONCEPT: Drug seeking behavior (DSB) is often mixed in illicit drug diversion confounding legitimate attempts to control acute and chronic pain. OBJECTIVE: To review the literature of acute and chronic pain control against the medical and legal context of DSB. DESIGN: Retrospective literature review from National Library of Medical Computerized Data Base 1990--2004. PATIENTS: Preference to human prospective on retrospective clinical trials. RESULTS: Drug use and abuse have significant adverse consequences. Pain control is desirable and necessary with chronic pain syndromes more prone to DSB. This behavior can be accurately profiled and information used to assist recovery. CONCLUSION: It is desirable to address DSB stressing acceptance and a multidisciplinary approach to recovery. PMID- 15540495 TI - Characteristics of adolescents in residential treatment for heroin dependence. AB - A retrospective review of the clinical charts of 97 adolescents who had received residential treatment for heroin dependence was conducted to determine predictors of heroin use. The average age for the adolescent patients reviewed was 17 years. Almost half of the adolescents (48%) were female and over half of the adolescents (53.6%) used heroin intravenously. Both males and females had substantial heroin habits, but differing amounts of use. Boys used an average of six bags of heroin per day and girls used an average of four bags of heroin per day. Using gender, age, number of heroin dependence symptoms, and other substances as predictors, 64.21% of the adolescents were correctly classified as injectors or noninjectors of heroin. Heroin appears to be a significant drug of abuse among these predominately white, middle class, suburban adolescents. PMID- 15540496 TI - Treatment retention predictors of drug court participants in a rural state. AB - Factors distinguishing clients who complete drug court treatment from those who do not complete drug court have been documented, but differences between urban and rural drug court participants have not been examined. The present study focuses on examining mental health, drug use, criminal activity, and education/employment as factors that are associated with treatment retention, which is measured by graduation from a rural and urban drug court. Study findings indicate that for the urban drug court, marital status, employment, drug use, and criminal activity predicted graduation. For the rural drug court, however, graduation was only predicted by age and juvenile incarceration. Findings from this study suggest there are different factors associated with drug court retention/graduation between urban and rural drug court settings. It is suggested that drug court administrators and other could use this information to better assess potential participants and to target services. PMID- 15540497 TI - Computerized screening of substance abuse problems in a primary care setting: older vs. younger adults. AB - The purpose of the current study was to examine differences in responses of older adults (age 55 and above) and younger adults (ages 18 to 54) to the Drug Abuse Problem Assessment for Primary Care (DAPA-PC), a computerized drug and alcohol abuse screening instrument developed for primary care settings. Data were collected from a diverse population of 327 adults presenting for care at The George Washington University Medical Faculty Associates clinic in downtown Washington, DC. Results indicated that rates of drug and alcohol abuse were similar in both groups. However, older adults were less likely than younger adults to perceive their drug use as problematic. This finding has serious implications for older adults, who tend to be underrepresented in treatment programs. There is a need for screening seniors and identifying those who may be at risk for substance abuse problems. Differences in responses to alcohol and drug assessments by age should be taken into consideration when designing screening instruments. The results of this study suggest that the DAPA-PC would provide a useful instrument for screening older adults in a primary care setting. PMID- 15540498 TI - Personal adjustment and substance abuse problems in a longitudinal study of TANF recipients and the potential need for treatment. AB - OBJECTIVE: This study assessed the need over time for treatment of personal adjustment and substance abuse problems among chronic drug using and other recipients of Temporary Assistance for Needy Families (TANF). METHODS: Participants were administered the personal adjustment problems and substance abuse problems scales contained in the Multidimensional Addictions and Personality Profile (MAPP) at intake, one-year, and two-year interviews. RESULTS: Most participants who were not chronic users had total scale scores that would indicate no recommended referral or a referral to outpatient treatment. Approximately one-third of chronic users had scores that would indicate referral to inpatient treatment. Many chronic users had comorbid conditions but reported receiving no psychological or substance abuse treatment. CONCLUSIONS: Many TANF recipients may be experiencing coexisting personal and substance abuse related problems that are going untreated. PMID- 15540499 TI - The short-term effects and unintended long-term consequences of binge drinking in college: a 10-year follow-up study. AB - This study addresses binge drinking in college as a risk factor for heavy drinking and alcohol dependence after college. A national probability sample of 1972 college students from the National Longitudinal Surveys of Youth (NLSY79) was interviewed in 1984 and reinterviewed again as adults in 1994. The short-term effects of binge drinking in college were assessed as well as the extent to which experiences of negative effects in college predicted patterns of alcohol use across the transition from college into postcollege years. As expected, college binge drinkers were comparatively more likely than nonbinge drinkers to experience one or more alcohol-related problems while in college. In addition, weighted estimates of DSM-IV-defined diagnostic criteria in logistic regression models indicated that the binge drinking patterns exhibited during the college years, for some former college students of both genders, posed significant risk factors for alcohol dependence and abuse 10 years after the initial interview, in conjunction with evidence of academic attrition, early departure from college and less favorable labor market outcomes. PMID- 15540500 TI - Lability and impulsivity synergistically increase risk for alcohol-related problems. AB - This study examined two aspects of affect dysregulation as risk factors for alcohol-related problems. From a sample of 592 undergraduates, 442 alcohol users were examined on measures of impulsivity, lability, alcohol use, and alcohol related problems. As expected, affect lability and impulsivity significantly increased risk for alcohol problems even after controlling for gender and lifetime-use frequency. Impulsivity was further examined as a vulnerability factor, potentially enhancing the relations between use frequency and problems and affect lability and problems. Impulsivity did not moderate the use frequency problems relationship. However, the relationship between affect lability and problems was greatest among participants with higher degrees of impulsivity. The results demonstrate that individual differences in affect dysregulation are associated with alcohol-related problems in young adults. PMID- 15540501 TI - Transfer to intermediate forms following concept discrimination by pigeons: chimeras and morphs. AB - Two experiments examined pigeons' generalization to intermediate forms following training of concept discriminations. In Experiment 1, the training stimuli were sets of images of dogs and cats, and the transfer stimuli were head/body chimeras, which humans tend to categorize more readily in terms of the head part rather than the body part. In Experiment 2, the training stimuli were sets of images of heads of dogs and cats, and the intermediate stimuli were computer generated morphs. In both experiments, pigeons learned the concept discrimination quickly and generalized with some decrement to novel instances of the categories. In both experiments, transfer tests were carried out with intermediate forms generated from both familiar and novel exemplars of the training sets. In Experiment 1, the pigeons' transfer performance, unlike that of human infants exposed to similar stimuli, was best predicted by the body part of the stimulus when the chimeras were formed from familiar exemplars. Spatial frequency analysis of the stimuli showed that the body parts were richer in high spatial frequencies than the head parts, so these data are consistent with the hypothesis that categorization is more dependent on local stimulus features in pigeons than in humans. There was no corresponding trend when the chimeras were formed from novel exemplars. In Experiment 2, when morphs of training stimuli were used, response rates declined smoothly as the proportion of the morph contributed by the positive stimulus fell, although results with morphs of novel stimuli were again less orderly. PMID- 15540502 TI - Matching: its acquisition and generalization. AB - Choice typically is studied by exposing organisms to concurrent variable-interval schedules in which not only responses controlled by stimuli on the key are acquired but also switching responses and likely other operants as well. In the present research, discriminated key-pecking responses in pigeons were first acquired using a multiple schedule that minimized the reinforcement of switching operants. Then, choice was assessed during concurrent-probe periods in which pairs of discriminative stimuli were presented concurrently. Upon initial exposure to concurrently presented stimuli, choice approximated exclusive preference for the alternative associated with the higher reinforcement frequency. Concurrent schedules were then implemented that gave increasingly greater opportunities for switching operants to be conditioned. As these operants were acquired, the relation of relative response frequency to relative reinforcement frequency converged toward a matching relation. An account of matching with concurrent schedules is proposed in which responding exclusively to the discriminative stimulus associated with the higher reinforcement frequency declines as the concurrent stimuli become more similar and other operants-notably switching-are acquired and generalize to stimuli from both alternatives. The concerted effect of these processes fosters an approximate matching relation in commonly used concurrent procedures. PMID- 15540503 TI - Contingencies of reinforcement in a five-person prisoner's dilemma. AB - As in studies of self-control, a tit-for-tat contingency in an iterated prisoner's dilemma game creates a conflict between maximization of local and global reinforcement. The present experiments examine this conflict in a multiplayer prisoner's dilemma game. Versus tit for tat, cooperation corresponds to self-control; defection, always immediately reinforced, corresponds to impulsiveness. Three experiments examined sensitivity of behavior to the global reinforcement contingency imposed by tit for tat. Undergraduates played a five player prisoner's dilemma game against four dummy players programmed to play tit for tat or randomly. With tit for tat, a player's cooperation (or defection) increased dummy players' cooperation (or defection) on subsequent trials reinforcing cooperation and punishing defection in the long run. Participants cooperated at a higher rate when the dummy players played tit for tat than when the dummy players played randomly. These results are consistent with findings in corresponding studies of self-control. Some participants, caught in a trap of mutual defection with the tit-for-tat playing dummy players, came to cooperate when the tit-for-tat contingency was reset ("forgiving" participants' previous defections) during a pause in the game. This increase was a result of the combined effects of a pause and reset; neither pausing nor resetting independently resulted in an increase in cooperation. PMID- 15540504 TI - The transformation of consequential functions in accordance with the relational frames of same and opposite. AB - Although the literature on reinforcement in behavioral psychology is extensive, few studies have examined the derived transformation of reinforcing functions in accordance with equivalence classes, and no published research has yet examined the derived transformation of consequential functions in accordance with nonequivalence relations. In the present study, which consisted of four experiments, the basic preparation was as follows. First, an arbitrary stimulus, B2, was established as a conditioned punisher, using direct stimulus pairing. Following nonarbitrary relational training, designed to establish SAME and OPPOSITE contextual cues, subjects were exposed to arbitrary relational training using these contextual cues to establish A1 as the same as B1 and C1, and as opposite to B2 and C2. Subsequently, C2 (based on its Same relation with B2) functioned as a punisher and C1 (based on its Opposite relation with B2) functioned as a reinforcer in a simultaneous discrimination task. Critically, the C1 stimulus acquired reinforcing functions, based on the derived relation of Opposite, although no such function had actually been established for any member of the network. Furthermore, these effects were observed across ABA reversals in the baseline contingencies. PMID- 15540505 TI - The effects of morphine on the production and discrimination of interresponse times. AB - Recent experiments suggest that the effects of drugs of abuse on the discrimination of the passage of time may differ for experimenter-imposed and subject-produced events. The current experiment examined this suggestion by determining the effects of morphine on the discrimination of interresponse times (IRTs). Pigeons pecked a center key on a random-interval 20-s schedule of matching-to-sample trials. Once the interval had timed out, a choice trial randomly followed either a short (2- to 3-s) or long (6- to 9-s) IRT on the center key. Pecking the side key lit one color produced food after a short IRT, and pecking the side key lit the other color produced food after a long IRT. Two experimental phases differed in the functional role of the different key colors. Under control conditions, the IRT distributions had two modes, one at the lower bound of the short category and a smaller one at the lower bound of the long category. Pigeons accurately categorized the duration of the IRTs: One key color was pecked following short IRTs and the other key color was pecked following long IRTs. Morphine flattened the IRT distribution and reduced the accuracy of categorizing IRTs. Categorization of long IRTs was particularly disrupted. Morphine did not produce overestimation of time as assessed by the production or categorization of IRTs. These results are similar to those obtained previously for the effects of morphine on the discrimination of the duration of experimenter imposed events. PMID- 15540506 TI - Relational operants: processes and implications: a response to Palmer's review of Relational Frame Theory. AB - Palmer has recently criticized Relational Frame Theory (RFT) on the grounds that it has developed data in search of a principle. In this reply, we show that he has done so by attacking fundamental concepts within behavior analysis itself, including the functional nature of an operant and contingencies of reinforcement as a behavioral process. His claim that RFT appeals to new behavioral principles to explain the development of relational operants is shown to be incorrect: As with any operant, RFT appeals to a history of contacted consistencies in contingencies across multiple ex-emplars to explain them. New principles only emerge later as a logically necessary extension of such operants if they exist--a view that Palmer failed to address or appreciate. Palmer's desire to see the use of methods other than matching-to-sample is proper but already largely satisfied in the empirical literature on RFT. We show Palmer's defense of Skinner's definition of verbal behavior to be illogical and unresponsive to the empirical challenge behavior analysis faces. Palmer's alternative common sense mediational associationistic account is another in more than a century of such accounts, all of which have failed empirically. At its root, Palmer's criticism is based on a mechanistic philosophy that is hostile to a traditional functional behavior analytic approach. PMID- 15540507 TI - Coagulopathies and the kidney. PMID- 15540508 TI - Antiphospholipid antibodies and hypertension. AB - Hypertension is a common manifestation of antiphospholipid syndrome (APS). Antiphospholipid antibodies (aPL) have been described in patients with hypertension secondary to renal artery stenosis (RAS). Twenty-six patients with RAS and 25 patients with severe essential hypertension (diastolic blood pressure > 110 mmHg or > or = 3 hypertensive drugs) were studied and compared to 61 age- and sex-matched healthy subjects. Serum samples were tested for lupus anticoagulant (LA), anticardiolipin (aCL) IgG and IgM, antiprothrombin (aPT) IgG and IgM, anti-beta2glycoprotein 1 (abeta2GP1) IgG and IgM. aPL were negative in all patients with RAS. Two patients with essential hypertension had positive aPL (8%) (LA in one patient confirmed in a second assay and abeta2GP1-IgG in the other patient confirmed one year later together with aCL IgG positivity). Among healthy subjects, one case (1.6%) was found to be positive for LA, aCL IgM, abeta2GP1 IgM, aPT IgG, aPT IgM. In conclusion, the association between RAS and aPL seems to be casual rather than an expression of an elective thrombotic localization ofAPS. The positive finding of aPL in 8% of patients with essential hypertension, a frequency higher than that of the control population, deserves further studies in larger series to better explore the relationship between aPL and hypertension. PMID- 15540509 TI - Epidermal growth factor receptor (EGFR) gene Bsr I polymorphism is associated with systemic lupus erythematosus. AB - The purpose of this study was to determine if epidermal growth factor receptor (EGFR) gene polymorphism was a marker of susceptibility to or severity of Chinese patients with systemic lupus erythematosus (SLE) in Taiwan. The study included 119 Chinese patients with SLE. One hundred unrelated healthy individuals living in central Taiwan served as control subjects. Polymorphisms of the EGFR Bsr I gene were typed from genomic DNA. The genotypes, allelic frequencies and carriage rates were compared between SLE patients and control subjects. The relationship between allelic frequencies and clinical manifestations of SLE was evaluated. For the genotype of EGFR gene Bsr I polymorphism, there was statistically significant differences between the SLE and control groups (chi-squared test, P = 0.009, chi2 = 9.21). In addition, there was significant association between the two groups in allelic frequency of the T allele (P = 0.02, chi2 = 5.27). However, we did not detect any association between EGFR genotype and clinical or laboratory profiles in SLE patients. The results suggest that the EGFR gene Bsr I polymorphism is related to SLE. PMID- 15540510 TI - Hypocomplementemia in systemic lupus erythematosus and primary antiphospholipid syndrome: prevalence and clinical significance in 667 patients. AB - The objective of the study was to analyse the prevalence and clinical significance of hypocomplementemia in a large series of patients diagnosed either with systemic lupus erythematosus (SLE) or with primary antiphospholipid syndrome (APS) and its association with the main clinical, hematological and immunological features of these diseases. Between 1992 and 2003, complement determinations (C3 and C4 levels, CH50 activity) were performed in 597 consecutive patients diagnosed with SLE (530 women and 67 men, mean age 32.6 years) and 70 with primary APS (57 women and 13 men, mean age 38.7) visited in our department. Complement determinations are routinely made at the first visit of patients and yearly during the follow-up. SLE and primary APS were diagnosed according to current classification criteria. Hypocomplementemia was detected in 371 (62%) of SLE patients. Compared with patients with normal complement values, those with hypocomplementemia showed a higher prevalence of female gender (P < 0.001), fever (P = 0.021), nephropathy (P < 0.001), cutaneous vasculitis (P = 0.023), positive anti-dsDNA antibodies (P = 0.012) and cryoglobulinemia (P < 0.001). In addition, patients with hypocomplementemia showed a higher prevalence of APS-related features such as hemolytic anemia (P = 0.001) and antiphospholipid antibodies (P < 0.001). Hypocomplementemia was prospectively related to accumulated hospitalization in SLE patients but not with the accumulated number of lupus flares or with the survival after follow-up of five years. In contrast, 33 (47%) patients with primary APS presented low complement values, which were associated with a higher prevalence of livedo reticularis (P = 0.022), thrombocytopenia (P = 0.004), lupus anticoagulant (P = 0.013), positive IgM-aCL (P = 0.039), positive ANA (P = 0.002) and anti-dsDNA (P = 0.046). The diagnostic value of hypocomplementemia in patients with SLE is based on the association with disease activity, immune-complex mediated manifestations (glomerulonephritis, cryoglobulinemia) and APS-related features (livedo reticularis, hemolytic anemia and aPL). Hypocomplementemia was found in nearly half of patients with primary APS, and was associated with some APS features (livedo reticularis, thrombocytopenia, aPL) but also with SLE-related immunological markers (ANA and anti-dsDNA), identifying a subset of patients with primary APS with a higher risk of evolving to SLE. These results clearly support the routine determination of complement factors in the clinical follow-up of patients with SLE and primary APS. PMID- 15540512 TI - Autoantibodies and self-reported health complaints in relatives of systemic lupus erythematosus patients: a community based approach. AB - First-degree relatives (FDRs) and spouses to a population-derived cohort of lupus patients were investigated for the occurrence of selected autoantibodies and self reported health complaints. A healthy reference population was included. The lupus population consisted of 103 index cases. A total of 275/375 available relatives accepted to enter the study. Two hundred and twenty-six/315 (72%) were FDRs and 49/60 (82%) were spouses. Serum was analysed for ANA using indirect immunofluorescence on Hep-2 cells at the following dilutions: 1:40, 1:80 and 1:160 and in addition sera were tested for anti-dsDNA, IgM RF, ACA (IgM, IgG), anti-beta2GPI (IgM, IgG) and antibodies to prothrombin. ANA positivity occurred more frequently in FDRs compared with spouses and controls at serum dilution 1:160 (10 versus 0% and 2.5%, respectively, P = 0.04 and P < 0.01), 1:80 (24 versus 4% and 5%, respectively, P = 0.003 and P < 0.001) and 1:40 (31 versus 10% and 10%, respectively, P = 0.006 and P < 0.0001). ANA positivity in FDRs occurred randomly, irrespective of family relationship. Fifty-three/184 versus 2/32 FDRs to patients with definite SLE (D-SLE) and incomplete SLE (I-SLE), respectively, tested ANA positive at 1:80 (P < 0.05). FDRs with ANA titer at 80 were affiliated to lupus probands with high SLICC scores (P < 0.05). Self-reported health complaints, cardiovascular/thromboembolic events in particular, were more frequent among FDRs than in spouses. The population-based approach adopted in the present study supports previous clinic-based evidence of an increased propensity for autoantibody occurrence in relatives to SLE patients. In FDRs, present ANA positivity was associated with increased prevalence of health complaints and ANA positivity in FDRs was related to the criterial burden and cumulated damage in corresponding lupus probands. The low ANA frequency among spouses of SLE patients argues against a significant autoantibody triggering effect of shared environment in adult life. PMID- 15540511 TI - CTLA-4 gene polymorphisms and systemic lupus erythematosus in a population-based study of whites and African-Americans in the southeastern United States. AB - Cytotoxic lymphocyte antigen-4 (CTLA-4) plays an important role in regulating T cell activation, and may help to limit T cell response under conditions of inflammation. Genetic variability in CTLA-4 has been implicated in the development of several autoimmune diseases. Some studies have described associations between CTLA-4 polymorphisms and systemic lupus erythematosus (SLE), but findings have been inconsistent. We examined polymorphisms in the CTLA-4 gene promoter region (-1722T/C, -1661 A/G, -318C/T) and exon I (+49G/A) with respect to SLE in a population-based case-control study in the southeastern US. Genotypes from 230 recently diagnosed cases and 276 controls were examined separately for African-Americans and whites. We observed no overall associations between SLE and the four CTLA-4 polymorphisms examined. Subgroup analyses revealed effect modification by age for the presence of the -1661G allele, yielding a significant positive association with SLE in younger (<35 years) African-Americans (OR = 3.3). CTLA-4 genotypes also interacted with HLA-DR2 and GM allotype to contribute to risk of SLE. These findings suggest allelic variation in this region of CTLA4 is not a major independent risk factor for SLE, but may contribute to risk of disease in younger African-Americans or in the presence of certain immunogenetic markers. PMID- 15540513 TI - Autoantibodies recognizing the 27 carboxy-terminal amino acids of calpastatin are associated with secondary Sjogren syndrome in systemic lupus erythematosus. AB - The objective of this study was to determine in systemic lupus erythematosus (SLE) the prevalence and clinical significance of anticalpastatin antibodies (ACAST), an autoantibody population previously detected in sera from patients with various connective tissue diseases. Eighty-four patients with SLE (mean age: 30 years at diagnosis, females 77) that fulfilled ACR criteria were included in the study retrospectively. Several clinical and biological data were collected. ACAST were detected by a solid-phase enzyme linked immunosorbent assay (ELISA) using as antigen a synthetic peptide corresponding to the 27 C-terminal amino acids of calpastatin (CAST-C27). The prevalence of ACAST-C27 was 13% (11/84) in SLE patients. No correlation was found between the presence of ACAST-C27 and clinical manifestations such as thrombosis and vasculitis. Furthermore, no correlation was observed with the presence ofantiphospholipid antibodies (APL). However, we found a statistically significant association between the presence of ACAST-C27 and that of secondary Sjogren syndrome (P = 0.01). The conclusion is ACAST-C27 are not associated with thrombosis in SLE patients. The association observed between ACAST-C27 and secondary Sjogren syndrome suggests that ACAST-C27 might be useful in discriminating a clinical subgroup of SLE patients. PMID- 15540514 TI - Neuropsychological development of children born to patients with systemic lupus erythematosus. AB - To verify the neuropsychological development in the offspring of patients with systemic lupus erythematosus (SLE), 47 children (23 male and 24 female) from affected women were studied. The tests applied were related to the children's ages: Griffiths scale up to four years, WPPSI and metaphonological tests (MP, evaluating the phonological consciousness) from four to six years of age, WISC-R test and Rey test (evaluating the visual-space abilities) from six years onwards; finally, specific tests for the diagnosis of learning disabilities (LD) between the ages of seven and 13. Intelligence levels were always normal (mean IQ score 106.32; median 104; SD 9.05). Three out of eight examined children failed MP, therefore may develop LD and will need further evaluation later. Fourteen children were specifically studied for LD and three reported scores lower than normal, but only two (who were brothers) were defined dyslexic. Antiphospholipid antibodies (aPL) were positive in the mothers of the three children with impaired LD tests. Other maternal autoantibodies or drugs administered during pregnancy did not seem to be related to LD. In conclusion, maternal SLE does not impair intelligence levels, but may increase the occurrence of LD particularly in male children (2/8 males examined, 25%). Both maternal aPL and genetic background may have pathogenetic implications. PMID- 15540515 TI - Familial antiphospholipid antibodies and acquired circulating anticoagulants. AB - Familial clustering of elevated antiphospholipid antibody levels has been described, but the reports are heterogeneous with regard to the characterization of the antiphospholipid syndrome (APS), coexisting autoimmune diseases and clinical implications. We report a familial occurrence of APS in two patients, in the presence of SLE in the mother and absence of autoimmune diseases in the daughter along with acquired circulating inhibitors in both cases: platelet inhibitor and factor IX inhibitor, respectively. PMID- 15540516 TI - Discoid lupus erythematosus of the vulva. AB - Genital involvement is a rare manifestation in discoid lupus erythematosus (DLE) and systemic lupus erythematosus (SLE), and there are few reports on vulvar clinical features. We describe a 41-year old Caucasian woman with bilateral vulvar DLE. Although vulvar lesions in lupus are probably more common than is realized, the present report is the first case of genital DLE documented in a female. PMID- 15540517 TI - Respiratory arrest in systemic lupus erythematosus due to phrenic nerve neuropathy. AB - Diaphragmatic weakness in patients with systemic lupus erythematosus (SLE) is a controversial issue and is claimed to have a neuropathic, myopathic or unknown pathogenesis. In this patient a predominantly motor neuropathy with diaphragmatic paralysis due to axonal involvement of the phrenic nerve was discovered and successfully treated with immunosuppressive drugs. PMID- 15540518 TI - Systemic lupus erythematosus in Pakistan. AB - Clinical features of systemic lupus erythematosus (SLE) have been described from different geographical regions in the world, with some clinical differences among different racial groups. Although data on the characteristics of SLE in Pakistan is scarce, it is not uncommon in the South East Asian region. The purpose of this study was, therefore, to delineate the clinical pattern and disease course in Pakistani patients with SLE and to compare it with international data on lupus patients. A total of 196 patients with SLE fulfilling the clinical and laboratory criteria of the American Rheumatism Association admitted to the hospital between 1986 and 2001 were studied by means of a retrospective review of their records. Demographically, it was seen that SLE is a disease predominantly of females in their third decade, which is consistent with worldwide data. The mean age of presentation was 31 years (range 14-76) and the mean duration of follow up was 34 (4-179) months. Generally, there was less cutaneous (46%), arthritic (38%), serositis (22%) and renal involvement (33%) but more neuropsychiatric symptoms (26%) in our population. Eighty-six percent of patients were ANA positive, whereas anti dsDNA was positive in 74% of patients. Infections, renal involvement, seizures and thrombocytopenia were associated with poor prognosis (P < 0.05). This study is the first of its kind in Pakistan. The clinical and laboratory characteristics of SLE patients in our study place our population in the middle of a spectrum between the Caucasians and other Asian populations. It has shown that the clinical characteristics of SLE patients in this country may be different to those of its neighbors. PMID- 15540519 TI - Anetoderma associated with systemic lupus erythematosus and antiphospholipid antibodies. PMID- 15540520 TI - Teaching suturing in a workshop setting: a comparison of several models. AB - OBJECTIVES: Suturing is taught in workshops using a variety of models. The purpose of this study is to compare the resemblance to human skin of four models commonly used to teach suturing: pig skin, beef tongue, hot dog and latex glove. METHODS: 5 centimeter biconvex incisions were made in each of the models and closed by 50 physician study volunteers comprised of 33 board-certified physicians and 17 resident physicians. They rated each model on a scale of 1 to 4, where 4 closely resembles human skin and 1 does not resemble human skin. RESULTS: The following mean ratings were given by study volunteers: beef tongue 3.5 +/- 0.5, pig skin 3.2 +/- 0.8, latex glove 1.6 +/- 0.7, hot dog 1.4 +/- 0.6. CONCLUSION: Beef tongue and pig skin were rated highest by study volunteers. However, pig skin is much cheaper than beef tongue. Pig skin is the best inexpensive model for teaching skin suturing of the four models studied. PMID- 15540521 TI - Hemorrhagic fever with renal syndrome. AB - Hemorrhagic fever with renal syndrome (HFRS) is caused by the Hantaviruses, a group enveloped RNA viruses transmitted through contact with infected rodent urine or feces. Although distributed widely through Europe, Asia, and the New World, infections acquired in Korea, China, and Russia tend to be among the most severe. The initial presentation of HFRS is extremely variable, but generally includes fever, malaise, headache and abdominal pain. Laboratory findings that may lead to the diagnosis include thrombocytopenia, azotemia, elevated serum creatinine, or proteinuria. We present the case of a patient that acquired hemorrhagic fever with renal syndrome in South Korea. PMID- 15540522 TI - Hemophagocytic lymphohistiocytosis: a rare cause of pancytopenia. AB - A 36-year-old man with fever and pancytopenia due to Hemophagocytic Lymphohistiocytosis is reported. The patient was started on the HLH-94 based treatment. Two weeks after the initiation of therapy the patient's pancytopenia had resolved and he was discharged to complete treatment as an outpatient. The initial clinical presentation, diagnostic criteria, pathophysiology and treatment will be discussed. PMID- 15540523 TI - Effects of Upcountry Maui water additives on health. AB - Since 2001 Upcountry Maui residents have been concerned that water additives may be linked to health problems in their community. A study using phone surveys was conducted to assess this issue. Most people suffered skin rashes, while others experienced eye irritations or respiratory problems. The surveys suggested that these symptoms might have been attributable to the water additives. PMID- 15540524 TI - Acute renal failure resulting from intravenous immunoglobulin therapy. AB - Intravenous administration of immunoglobulin is used for the treatment of many conditions, including primary immunodeficiency states, autoimmune disorders, glomerulonephritides and polyneuropathy. Acute renal failure induced by intravenous immunoglobulin is a known but rare adverse reaction. We have a patient who was treated with IVIG for inflammatory polyneuropathy. Intravenous immunoglobulin therapy 0.5 g/kg/ d was given for 4 days. Three days after completion of IVIG therapy, patient developed decreased urine output. His serum creatinine increased from baseline of 1.3 to 7 mg/dL. Even though IVIG was discontinued, patient required hemodialysis. This case illustrated that IVIG can cause acute oliguric renal failure which is reversible after withdrawal of the drug. Risk factors include pretreatment renal impairment, diabetes mellitus, high concentration of sucrose or glucose in IVIG preparation and older age. Awareness of this serious side effects and recognition of predisposing factors provide means of avoiding a known life threatening complication of IVIG therapy. PMID- 15540525 TI - Comparing need between health occupation and health education schools: which students benefit most from the school health education program. AB - Comparing need between Health Occupation and Health Education Schools: Which students benefit most from the School Health Education Program? First-year medical students taught general health topics at public high schools. Pre-test and post-tests were given for each presentation. Health Education students had lower pre-test scores but showed greater improvement. With greater need and fewer resources, Health Education students benefit most. PMID- 15540526 TI - John A. Burns School of Medicine Class of 2008, profile. PMID- 15540527 TI - The Cancer Biomedical Informatics Grid (caBIG): pioneering an expansive network of information and tools for collaborative cancer research. PMID- 15540528 TI - [Threat or creation of panic? Disease agents as terror weapons]. PMID- 15540529 TI - [When is osteoporosis prophylaxis necessary! Sick intestine consumes the bones]. PMID- 15540530 TI - [New test identifies tumor markers in stool. Can colonic carcinomas now be detected earlier?]. PMID- 15540531 TI - [A patient with fibrotic diathesis and linitis plastica. Is there a relationship between those diseases?]. PMID- 15540532 TI - [Apoplexy doubles the risk of dementia]. PMID- 15540533 TI - [Osteopathy in tennis elbow?]. PMID- 15540534 TI - [Diabetology and cardiology--a hearty association]. PMID- 15540535 TI - [Screening, prevention and early therapy of type 2 diabetics]. AB - For each known diabetic at least one new patient is diagnosed when the 75-g OGTT is used for screening. For early diagnosis in high risk groups, the oral glucose tolerance test is indispensable since ca. 30% of the asymptomatic diabetes cases are identified on the basis of a 2-hour postchallenge glucose value of > 11.1 mmol/l. In controlled studies on persons with IGT, every second diabetes case could be prevented through lifestyle modifications beginning in childhood. Three year studies showed that the incidence of diabetes could be reduced by about one third with the medications acarbose, metformin and orlistat. The observation that statins, ACE inhibitors and AT1 blockers also contributed to a reduction of new diabetes cases was made in studies on the prevention of coronary heart diseases. PMID- 15540536 TI - [Inflammation and diabetes]. AB - The focus of current diabetes research is the clarification of the pathogenetic relationships between subclinical inflammation, diabetes and arteriosclerosis. Even minimal disturbances in glucose tolerance are associated with a chronic, generalized inflammatory reaction that links components of the metabolic syndrome and contributes to the development of diabetic complications as well as to the development and progression of arteriosclerosis. The most important mediators and markers of this inflammation cascade are NF-kappaB, TNF-alpha, IL-6, CRP and PAI 1. For the treatment of subclinical inflammation, substances with anti inflammatory properties such as statins or ACE inhibitors are of increasing importance. PMID- 15540537 TI - [Cardiovascular risk in diabetes--diagnostic and therapeutic aspects]. AB - In Germany, more than 27,000 diabetic patients suffer an acute myocardial infarction annually; in addition, the number of cardiac infarction patients with unrecognized permanent disturbances of the glucose metabolism is high. The cardiovascular mortality of diabetics in comparison with nondiabetics is four to six times higher. Through an intensification of diagnostics and therapy, it is possible to reduce the hospital mortality of diabetics with acute myocardial infarction. For each patient with myocardial infarction and for whom diabetes has not been previously diagnosed, it is recommended that the blood sugar be tested upon admission to the hospital and that an oral glucose tolerance test be conducted for the detection of a glucose metabolic disorder during the postinfarction phase. An example of a successful approach for metabolic intervention is glucose-insulin infusion to optimize the metabolic condition during the acute phase. PMID- 15540538 TI - [Aerobilia as an initial sign of a cholecystoduodenal fistula--a case report]. AB - An 82 year old female patient has the following medical history: repeated chemotherapy and radiation therapy for non-Hodgkin's lymphoma, a radiation induced osteoblastic osteosarcoma and a longstanding cholecystolithiasis with sclero-atrophic cholecystitis. She appeared at the practice and complained about a dull food-dependent mesogastric and epigastric pain as well as a discrete scleral jaundice. Sonography and CT showed air inclusions in the bile ducts. Subsequently, a cholecystoduodenal fistula was found intraoperatively. The cause could be the long-existing cholecystolithiasis. PMID- 15540539 TI - [Treatment and prophylaxis of an acute migraine attack]. AB - The treatment of migraine takes into consideration the intensity of the headache and the accompanying symptoms. The goal is to reduce the headache intensity, to relieve nausea and vomiting and to restore the ability to function in the daily routine. For drug treatment, there are now highly effective serotonin agonists available in addition to the older ergotamine preparations. A problem with all migraine analgesics is that in long lasting migraine attacks, the headache can recur as the pharmacological effect of the medication wears off. Through nonmedicinal and medicinal prophylactic measures, the frequency and intensity of the migraine attacks can be reduced. Keeping a headache diary for documentation is essential. PMID- 15540540 TI - [Ulla's health professional card is bound to help insurance. And the physicians must pay for it]. PMID- 15540541 TI - [Health care reform shows effects. Insurance makes profits]. PMID- 15540543 TI - [Small estimate by expert witness written to insurance. Therewith you can be exposed to publicity]. PMID- 15540542 TI - [A firm dissatisfied with disability certificates. The physician nevertheless is not to be excluded]. PMID- 15540544 TI - [Lowering of cholesterol. What role is played by the choice of statins?]. PMID- 15540545 TI - [The 10-minute consultation: accidently discovered hypercholesteremia]. PMID- 15540546 TI - [Look diagnosis: Fingers at first blue then black]. PMID- 15540547 TI - [Cough, asthma, pneumonia, sleep apnea ... Airways react sour to acid reflux]. PMID- 15540548 TI - [When GERD "becomes odd". Antacids help persistent cough]. PMID- 15540549 TI - [Remarkably many asthma patients have reflux. Asthma--a case for PPI?]. PMID- 15540550 TI - [Reflux in intensive care patients. Now threatens pneumonia]. PMID- 15540551 TI - [Cravats as bacteria sling. Bacilli with necktie]. PMID- 15540552 TI - [Continued education for physicians in clinical weekday. When were you in the OR last?]. PMID- 15540553 TI - [Interactive and near-practice continuing education. How do the clinicians appear in the cards?]. PMID- 15540554 TI - [Somatization syndrome: a diagnostic and therapeutic challenge. Difficult patients with puzzling complaints]. PMID- 15540555 TI - [Irritable bowel syndrome: the challenge to psychosomatic and internal medical specialists to collaborate closely]. AB - The irritable bowel syndrome is a ubiquitous disease that usually exhibits a close affinity to the group of somatoform disorders. Chronic cases in particular often suffer from psychological disturbances, and consult numerous different medical, mainly internistic-gastroenterological, treatment centers, not infrequently with no lasting success. Although the importance of gastroenterological pathophysiological aspects is beyond doubt, an early interdisciplinary diagnostic work-up--with explicit involvement of the psychosomatic-psychotherapeutic specialty--is urgently recommended. Against the background of moderate concomitant medication, a psychosomatic-psychotherapeutic treatment regimen has proven successful. PMID- 15540556 TI - [Chronic fatigue syndrome]. AB - The symptoms of fatigue and exhaustion, which are central to chronic fatigue syndrome and neurasthenia, can lay claim to being"psychopathological entities" in their own right. Although ubiquitous, they by no means reflect a homogeneous clinical picture. Chronic fatigue may occur after physical illness, such as viral infections, or be associated with a range of different psychiatric disorders. There is wide overlap with affective, anxiety, somatoform, and personality disorders, and this should be borne in mind when it comes to classifying these symptoms. To investigate the etiopathogenesis, multifactorial models are required. For treatment, adapted cognitive-behavioral therapy, possibly accompanied by an antidepressant, is effective. PMID- 15540557 TI - [The fibromyalgia syndrome]. AB - The Fibromyalgia Syndrome Fibromyalgia is a "pain disorder" and is diagnosed in accordance with the criteria of the American College of Rheumatology. The clinical presentation involves a complexity of symptoms with pains occurring at numerous sites identified by "tender points", disturbed sleep, increased fatigue, and numerous psychovegetative and psychic symptoms. Epidemiological data reveal high prevalence figures for fibromyalgia, and the disease is usually chronic. The etiopathogenesis can be described only within a multifactorial model. Numerous neurobiological and psychosocial factors militate in favor of a central disturbance of stress coping and pain perception. Multimode therapeutic approaches need to be individualized, but in many cases continue to be only moderately successful. PMID- 15540558 TI - [Pain and neuroborreliosis]. AB - Pain is one of the presenting symptoms in acute neuroborreliosis. Classically, acute neuroborreliosis--also known in Europe as Bannwarth's syndrome--is a combination of radicular pain, cranial neuritis and peripheral radiculitis and inflammatory changes of the CSF. The prognosis following antibiotic therapy is favorable. At least in its early stages, however, the diagnosis neuroborreliosis might be missed or mistaken. Thus, targeted assessment of typical signs is needed to expedite examination of the CSF which then permits definitive diagnosis. PMID- 15540559 TI - [Controlled clinical testing of an antidiarrheal]. PMID- 15540560 TI - [A general practitioner has reduced to the most needed--and is happy with it. So functions the assistant-free practice]. PMID- 15540561 TI - [Should Disease Management Programs be diversified so narrowly?]. PMID- 15540562 TI - [Disability certificates from 5 different physicians. Immediate notice!]. PMID- 15540563 TI - [2-year data of large clinical comparative studies. Type 2 diabetes: lasting metabolic control with pioglitazone]. PMID- 15540564 TI - [Warning signals insulin resistance. Insulin resistance causes not only diabetes]. PMID- 15540565 TI - [The 10-minute consultation. Urinary incontinence in a woman]. PMID- 15540566 TI - [Appearance diagnosis. Facial shingles]. PMID- 15540567 TI - [Birthday of a "potency pill". Helmut Newton celebrates posthumously likewise]. PMID- 15540568 TI - "The 4Rs of leadership". PMID- 15540569 TI - History. PMID- 15540570 TI - Health care for children in foster care. AB - Every month 6,600 children in Oklahoma live under the custody of the state, most as result of being abused or neglected by their own families. The state provides medical care to these children via the Medicaid program. The American Academy of Pediatrics(AAP) has set forth a guideline for optimal care of these children. We discuss the current Oklahoma health care system for foster children and suggest changes that may move Oklahoma in the direction of the AAP recommendations. A more uniform, organized medical system may not only meet a foster child's medical needs but may also provide a degree of continuity to an otherwise discontinuous process. PMID- 15540571 TI - Middle colic artery rupture in a patient with ANCA associated vasculitis: a case report. AB - Antineutrophil cytoplasmic antibody (ANCA) positive vasculitides are a heterogeneous group of disorders. They can be disabling multi-organ system conditions. Involvement of small and occasionally medium-sized blood vessels is generally the rule. Recent classification changes and advances in laboratory testing have made it easier to work up a patient suspected of having an ANCA associated vasculitis. Syndromes can at times overlap, leaving a class of patient that does not fit into one specific group. Patients frequently present with a myriad of symptoms ranging from sore throat, fever, athralgias and myalgias, to overt hemorrhage and/or renal failure. The standard treatment remains immunosuppression with steroids, as well as the use of cyclophosphamide, azathioprine, or methotrexate. Plasmapheresis and intravenous immunoglobulin (IVIG) are also used. Gastrointestinal complications are a rare manifestation, first described in polyarteritis nodosa and more recently noted in Wegener's granulomatosis. This paper describes a patient with ANCA positive vasculitis who had a spontaneous rupture of the middle colic artery and multiple superior mesenteric aneurisms. It reviews current literature on the ANCA positive vasculitides, including current classification and treatment modalities. PMID- 15540572 TI - The Oregon Health Plan. AB - The OHP has been successful in its goal of decreasing the number of uninsured in the state of Oregon and providing them with quality medical care at an affordable rate. When the plan started, nearly one-third of those individuals falling at or below the federal poverty level had no health insurance, and the plan has provided for these individuals. Furthermore, approximately 18% of all Oregonians had no health insurance in 1990. In 1997, the uninsured population fell to about 8%3. Also, the prioritization list has established an effective basic Medicaid benefit package that could be used by other state Medicaid programs and by insurance companies. Also, the use of Managed Care Plans and setting priorities for health care services has made the cost of health care for more customers affordable. The OHP is not perfect and will always be in a constant state of flux, because of changes in the financial state of Oregon and/or changes in the health care needs of the state. Therefore, the OHP will need to be continually evaluated on periodic bases for any needed changes. However, the state of Oregon should be commended for their attempt to explore uncharted waters, and to try to find an answer to the Nation's biggest health care concern/problem, which is to provide medical coverage for everyone in an affordable manner. PMID- 15540573 TI - Is Oklahoma the weak link? The decline in vaccine coverage levels in Oklahoma. AB - Oklahoma has demonstrated a decline in vaccine coverage levels in children over the past two years, moving from a national ranking of 31st in 2000 to 48th in 2002. This decline has prompted a review of available evidence to identify approaches to move the state toward national averages and ultimately to the Healthy People 2010 Objectives. Oklahoma is one of nine states that are well below the established goal of 80% vaccine coverage for children. Oklahoma is near the bottom of this group with an estimated level of only 65%, placing not only Oklahomans at risk but the nation as well. The following is a plan aimed at both the health department and all health care professionals. Indeed, without physician involvement, the situation will not improve. PMID- 15540574 TI - Thank you for your editorial in the July edition of the journal. PMID- 15540575 TI - Megacities and atmospheric pollution. PMID- 15540576 TI - Urban air pollution monitoring and correlation properties between fixed-site stations. AB - The rich regional air-monitoring network of the Emilia-Romagna region of Italy has been used to quantify the spatial variability of the main pollutants within urban environments and to analyze the correlations between stations. The spatial variability of the concentrations of the majority of pollutants within the city was very high, making it difficult to differentiate and characterize the urban environments and to apply legal limits with uniform criteria. On the other hand, the correlations between the fixed-site monitoring stations were high enough for their data to be retained generally very appropriately for controlling temporal trends. Starting from the high correlation level, a procedure was proposed and tested to derive pollution levels, using short-term measurements, such as passive samplers and mobile-station data. The importance of long-term statistics in urban air pollution mapping was emphasized. Treatment of missing data in time series and quality assurance were indicated as possible fields for applications for the correlation properties. PMID- 15540577 TI - Regulatory off-gas analysis from the evaporation of Hanford simulated waste spiked with organic compounds. AB - After strontium/transuranics removal by precipitation followed by cesium/technetium removal by ion exchange, the remaining low-activity waste in the Hanford River Protection Project Waste Treatment Plant is to be concentrated by evaporation before being mixed with glass formers and vitrified. To provide a technical basis to permit the waste treatment facility, a relatively organic-rich Hanford Tank 241-AN-107 waste simulant was spiked with 14 target volatile, semi volatile, and pesticide compounds and evaporated under vacuum in a bench-scale natural circulation evaporator fitted with an industrial stack off-gas sampler at the Savannah River National Laboratory. An evaporator material balance for the target organics was calculated by combining liquid stream mass and analytical data with off-gas emissions estimates obtained using U.S. Environmental Protection Agency (EPA) SW-846 Methods. Volatile and light semi-volatile organic compounds (<220 degrees C BP, >1 mm Hg vapor pressure) in the waste simulant were found to largely exit through the condenser vent, while heavier semi-volatiles and pesticides generally remain in the evaporator concentrate. An OLI Environmental Simulation Program (licensed by OLI Systems, Inc.) evaporator model successfully predicted operating conditions and the experimental distribution of the fed target organics exiting in the concentrate, condensate, and off-gas streams, with the exception of a few semi-volatile and pesticide compounds. Comparison with Henry's Law predictions suggests the OLI Environmental Simulation Program model is constrained by available literature data. PMID- 15540578 TI - Laboratory characterization of modified tapered element oscillating microbalance samplers. AB - Laboratory tests with generated aerosols were conducted to test the efficacy of two recent design modifications to the well-established tapered element oscillating microbalance (TEOM) continuous particulate matter (PM) mass monitor. The two systems tested were the sample equilibration system-equipped TEOM monitor operating at 30 degrees C, which uses a Nafion dryer as part of the sample inlet, and the differential TEOM monitor, which adds a switched electrostatic precipitator and uses a self-referencing algorithm to determine "true PM mass." Test aerosols included ammonium sulfate, ammonium nitrate, sodium chloride, copper (II) sulfate, and mixed aerosols. Aerosols were generated with an atomizer or a vibrating orifice generator and were equilibrated in a 450-L slow flow chamber before being sampled. Relative humidity in the chamber was varied between 10 and 90%, and step changes in humidity were executed while generating aerosol to test the response of the instruments. The sample equilibration system-equipped TEOM monitor does reduce, but not totally eliminate, the sensitivity of the TEOM mass monitor to changes in humidity. The differential TEOM monitor gives every indication of being a very robust technique for the continuous real-time measurement of ambient aerosol mass, even in the presence of semi-volatile particles and condensable gases. PMID- 15540579 TI - Long-term field characterization of tapered element oscillating microbalance and modified tapered element oscillating microbalance samplers in urban and rural New York State locations. AB - Long-term field comparisons of continuous and integrated filter measurements of mass concentrations of particulate matter (PM) with an aerodynamic diameter less than or equal to 2.5 microm (PM2.5) were performed at rural and urban sites in New York State. Two versions of the continuous tapered element oscillating microbalance (TEOM) mass monitor are deployed at each site, in addition to Federal Reference Method filter samplers. Data are grouped into monthly averages to retain and demonstrate seasonal differences. Strong seasonal dependence is observed-the TEOM monitors with the heated sensors are biased systematically low with respect to the Federal Reference Method measurements during the cold season. For the rural site, the average bias for the sample equilibration system (SES) equipped and standard TEOM monitors is 14 and 24%, respectively. At this location, the TEOM monitor measurements were biased low for all 34 months. For the urban site, the average bias for the SES and standard TEOM monitors is 8 and 18%, respectively. At this location, the TEOM monitor measurements are as likely to be biased high as low during the warm-season months. The hour averaged data from the two versions of the TEOM monitor are also compared, and also indicate that the SES-equipped version of the TEOM monitor captures 7-11% more PM2.5 mass at these locations. PMID- 15540580 TI - Modeling of gas-phase photodegradation of chloroform and carbon tetrachloride. AB - The relationship between the irradiance in a photoreactor and the rate of photodegradation of organics is essential in the scaling-up of photoreactors to treat large volumes of air contaminated with organic pollutants. In this study, the analysis is adopted to compare results obtained from two different photoreactors. Initially, the applicability of two light models in calculating the irradiance in two photoreactors was evaluated. Thereafter, kinetic models of ultraviolet (UV) photooxidation of chloroform (CHCl3) and carbon tetrachloride (CCl4) from the archived literature were tested using experimental data under various operating conditions and different irradiances. Sensitivity analyses were conducted using different values of model parameters to determine the significance of each parameter on the photodegradation of the two chlorinated organics. For compounds that undergo photolysis as a primary mode of degradation, the rate of photodegradation at low initial concentrations can be predicted easily by the following equation: d[C]/dt = -2.303Iave, lambdaepsilonlambdaphilambda[C]. Although the photodegradation of chlorinated organic compounds in dry and humid air can be predicted well, it is difficult to predict the Cl* sensitized oxidation occurring at high initial concentrations. A good agreement between the simulated and experimental data provides a sound basis for the design of large-scale reactors. PMID- 15540581 TI - Evaluation of vacuum and wet-wipe methods for removal of World Trade Center dust from indoor environments. AB - The destruction of the buildings at the World Trade Center (WTC) complex dispersed dust and debris into the surrounding area. Pulverized building material made up most of the fallout and was intermixed with combustion byproducts of the aircraft and building contents. A study was conducted to confirm the effectiveness of several cleaning procedures in removing WTC-related contamination from lower Manhattan residences. The contaminants included asbestos, lead, dioxin/furans, synthetic vitreous fibers (fibrous glass), and crystalline silica. All cleaning procedures included the common elements of vacuuming and wet wiping with soap and water. This cleaning procedure combination is effective in reducing WTC-related contamination to below health-based benchmarks. Post-cleaning environmental test results suggested that airborne asbestos measurements can be used as a surrogate parameter in clearance determinations. PMID- 15540582 TI - A simple model for estimating emissions of volatile organic compounds from grass and cut grass in urban airsheds and its application to two Australian cities. AB - Grass, and particularly cut grass, recently has been shown to emit significant amounts of volatile organic compounds (VOCs) into the atmosphere. Some components of these emissions are highly reactive and may contribute to photochemical smog in urban areas. A simple model for estimating the VOC emissions from grass and for grass cutting that allows these processes to be included in urban/regional emissions inventories is presented here. Using previous work and recent literature values, estimates are made of these biogenic volatile organic compound (BVOC) emissions for two typical urban airsheds, those including the cities of Sydney and Melbourne in Australia. Grass and cut grass could contribute approximately 2% for Sydney and 3% for Melbourne of the total VOCs emitted into these urban atmospheres annually. These contributions could rise to 4 and 5%, respectively, during the weekends of the summer growing season and, thus, could contribute to weekday/weekend ozone differences. It is recommended that the emissions of BVOCs from grass and cut grass be included in urban and global emissions inventories so that more accurate predictions of smog chemistry can be determined. PMID- 15540583 TI - Ozone measurements in South Carolina using passive samplers. AB - Passive samplers with two different collection substrates were used to obtain an average ozone concentration for 1 month during the summer of 2002 for each South Carolina county. One sampler contained a filter coated with indigo carmine, whose color fades when exposed to ozone. The fading was measured by reflectance spectroscopy. The other sampler contained filters that were coated with nitrite, which is oxidized to nitrate when exposed to ozone. The nitrate was measured by ion chromatography. Calibration curves were developed for the two methods by comparing color fading from indigo carmine and nitrate ion concentration from the nitrite filter with ambient ozone concentration measured by a co-located reference continuous UV ozone analyzer. These curves were used to calculate integrated ozone concentrations for samplers distributed across South Carolina. Using the indigo carmine method, the average ozone concentrations ranged from 21 to 64 ppb (average = 46 +/- 7.9 ppb, n = 58) across the 46 counties in the state during one summer month of 2002. Concentrations for the same time period from the nitrite-coated filters ranged from 23 to 62 ppb (average = 41 +/- 8.1 ppb, n = 58). Also for the same time period, the 23 continuous UV photometric ozone monitors operated by the South Carolina Department of Health and Environmental Control at sites within 10 miles of some of the passive monitors showed ozone concentrations ranging from 28 to 50 ppb (average = 39 +/- 6.3 ppb, n = 22). PMID- 15540584 TI - Photochemical smog modeling for assessment of potential impacts of different management strategies on air quality of the Bangkok Metropolitan Region, Thailand. AB - A photochemical smog model system, the Variable-Grid Urban Airshed Model/Systems Applications International Mesoscale Model (UAM-V/SAIMM), was used to investigate photochemical pollution in the Bangkok Metropolitan Region (BMR). The model system was first applied to simulate a historical photochemical smog episode of two days (January 13-14, 1997) using the 1997 anthropogenic emission database available at the Pollution Control Department and an estimated biogenic emission. The output 1-hr ozone (O3) for BMR, however, did not meet the U.S. Environmental Protection Agency suggested performance criteria. The simulated minimum and maximum O3 values in the domain were much higher than the observations. Multiple model runs with different precursor emission reduction scenarios showed that the best model performance with the simulated 1-hr O3 meeting all the criteria was obtained when the volatile organic compound (VOC) and oxides of nitrogen (NOx) emission from mobile source reduced by 50% and carbon monoxide by 20% from the original database. Various combinations of anthropogenic and biogenic emissions in Bangkok and surrounding provinces were simulated to assess the contribution of different sources to O3 pollution in the city. O3 formation in Bangkok was found to be more VOC-sensitive than NOx-sensitive. To attain the Thailand ambient air quality standard for 1-hr O3 of 100 ppb, VOC emission in BMR should be reduced by 50-60%. Management strategies considered in the scenario study consist of Stage I, Stage II vapor control, replacement of two-stroke by four-stroke motorcycles, 100% compressed natural gas bus, 100% natural gas-fired power plants, and replacement of methyltertiarybutylether by ethanol as an additive for gasoline. PMID- 15540587 TI - Remembering Jessie Ferne (1910--2004). AB - A biography and record of the times and achievements of Jessie Ferne Young, founder of the Canadian Association of Neuroscience Nurses. PMID- 15540588 TI - How the brain recognizes and responds to shock. AB - The rat that wishes to reach the goal of his learning and attain the reward, his very own mountain of cheese, must be able to navigate through the maze (Figure One). So the nurse, to reach the goal of understanding why the outward manifestations of shock occur must learn to navigate through the physiological maze of the body. After considering some general information on shock, we will embark on a tour through the maze created by the shock syndrome. PMID- 15540589 TI - Analysis of nursing assessments in a cohort patients with ruptured cerebral aneurysms. AB - Patients admitted with subarachnoid hemorrhage are monitored for symptoms of vasospasm. A prospective study was designed to compare two monitoring instruments: a standard neurological tool (SNR) and the National Institutes of Health Stroke Scale (NIHSS). The two assessment tools were compared to evaluate their concordance and to identify areas where efficiency in recording assessments might be improved. We found no statistical difference between the two tools in detecting symptomatic cerebral vasospasm. Substantial discrepancies in the documentation of observations were noted, particularly in the assessment of limb drift. Avoidance of these discrepancies may require further definition in the SNR tool. A qualitative component consisting of a review of the nurses' notes regarding neurological status in the patients' charts was conducted. It was demonstrated that nurses commonly document information in the progress notes that is already captured in the SNR. Further education of nurses in the use of assessment tools is therefore recommended to avoid redundancies and increase efficiency in recording clinical observations. PMID- 15540590 TI - The pyoverdins of Pseudomonas syringae and Pseudomonas cichorii. AB - The structure elucidation of the cyclic (lactonic) forms of the pyoverdins with a succinamide side chain originally produced by the closely related species Pseudomonas syringae and P. cichorii is reported. Mass spectrometry and nuclear magnetic resonance analyses as well as the determination of the configuration of the amino acids after degradation indicate that these two pyoverdins differ only by the replacement of the first in-chain serine by glycine. The pyoverdins of P. syringae and P. cichorii and the dihydropyoverdin of P. syringae can be used by both species as siderophores. PMID- 15540591 TI - Total, free and conjugated sterolic forms in three microalgae used in mariculture. AB - Total, free and conjugated forms (steryl esters, steryl glycosides and acyl steryl glycosides) of sterols from three microalgae that are extensively used in mariculture (Tetraselmis chuii, Nannochloropsis salina and Skeletonema costatum) were examined. The results revealed that cholesterol is the only common fraction detected in all investigated species and distributed in free and all conjugated forms. However, the total sterol content of T. chuii was about 325 microg/g dry wt, most of it was concentrated amongst 24-methylcholesta-5,24-diene-3beta-ol and 24-methylcholest-5-en-3beta-ol. On the other hand, the majority of the fractions were distributed in the free form. The total sterol content of N. salina was about 180 microg/g dry wt, cholesterol was the major fraction that was detected. Nevertheless, the dominant distribution forms were esterified. While in S. costatum, the total sterol content was 76 microg/g dry wt, approximately most fractions are quantitatively alike and dominated in the free form. Furthermore, our study shows clearly that most sterols are not distributed regularly within each form, a result that encouraged us to suggest a distribution of specific sterol fraction as a free or conjugated can be used as a serving tool in chemotaxonomic studies. PMID- 15540592 TI - Black flower coloration in wild Lisianthius nigrescens: its chemistry and ecological consequences. AB - The major pigments responsible for the flower color within the black flowered Gentianaceae, Lisianthius nigrescens, were characterized by HPLC and chemical analyses HPLC analysis showed one major and one minor anthocyanin and 3 major and 3 minor flavone glycosides. The anthocyanins [delphinidin-3-O-rhamnol(1 6)galactoside and its 5-O-glucoside] comprised an extraordinary 24% of the dry weight of wild collected L. nigrescens corallas, and were accompanied in a 1:1 ratio by a range of apigenin and luteolin 8-C-glucosides and their 7-O-methyl ethers. The high levels of anthocyanins and flavones (and their co-pigmentation) is thought to account for the almost complete absorption of both UV and visible wavebands observed by reflectance photography. PMID- 15540593 TI - Cuticular wax profiles of leaves of some traditionally used African Bignoniaceae. AB - Bignoniaceae, Newbouldia laevis, Markhamia acuminata, Spathodea campanulata and Kigelia africana were analysed by GC-MS. The principal constituents were represented by a homologous series of n-alkanes (C23-C33), n-alcohols (C18-C30) and related carboxylic acids (C16-C36). For N. laevis and M. acuminata, ursolic and oleanolic acid were the most abundant wax components (52 and 60%, respectively), followed by the C29, the C31 and the C33 n-alkanes. The predominant components of S. campanulata were n-alcohols (35%), with octacosanol and triacontanol as the most abundant ones, while K. africana is distinguished from these three members by the conspicuous absence of triterpenoic acids and the predominance of n-alkanes (70%) with hentriacontane and tritriacontane as the main representatives. Other notable constituents were sterols, albeit present in trace amounts. The wax profiles are discussed in terms of taxonomic characters. PMID- 15540594 TI - Rapid determination of volatile compounds emitted from Chimonanthus praecox flowers by HS-SPME-GC-MS. AB - A novel rapid, simple and solvent-free method was developed for determination of the volatile compounds from the flowers of Chimonanthus praecox Link using headspace solid-phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC-MS). The SPME conditions were firstly optimized and applied to sampling of the volatile compounds emitted from living Chimonanthus praecox L. flowers and excised Chimonanthus praecox L. flowers. Thirty-one compounds emitted from living flowers were identified for the first time, which mainly included 4 methyl-1,3-pentadiene (2.0%), a-phellandrene (4.7%), benzyl methanol (11.1%), trans-linalool oxide (furanyl ring) (5.3%), a-linalool (36.0%), methyl salicylate (24.5%) and acetic acid benzyl ester (5.9%). Comparing the emission from living flowers and excised flowers, twenty-eight compounds were found to be detected in the two emissions, and three compounds, n-pentadecane, n-cetane and n heptadecane, were only found in the emission from the living flowers, which shows that they might be biomarker compounds. PMID- 15540595 TI - Comparison of terpene composition in Engelmann spruce (Picea engelmannii) using hydrodistillation, SPME and PLE. AB - Terpenes emitted by conifer trees are generally determined by analysing plant extracts or essential oils, prepared from foliage and cones using steam distillation. The application of these procedures limits experiments to cut plant materials. Recently headspace techniques have been adopted to examine terpene emission by living plants. This paper deals with the application of solid-phase micro-extraction (SPME) for the analysis of terpenes emitted by conifers foliage of Engelmann spruce (Picea engelmannii), including its seedlings. The compositions of SPME extracts obtained for destroyed and non-destroyed old and juvenile spruce needles were compared with the compositions of essential oils and pressurised liquid extraction (PLE) extracts corresponding to the same plant materials. No substantial differences have been found in the qualitative terpene composition estimated by analysing essential oil and PLE and SPME extracts from non-destroyed old and juvenile foliage. The disintegration of spruce needles results in the formation of a significant amount of myrcene in the case of the old conifer foliage and non-terpenoic compounds in the case of juvenile conifer foliage. This phenomenon can be attributed to enzymatic reactions occurring in the destroyed plant cells. PMID- 15540596 TI - Separation of flavonoids from Aleurites moluccana leaves using chitosan modified with heptaldehyde. AB - Heptaldehyde-modified chitosan (heptyl-chitosan, CH-Hp) was investigated as adsorbent for chromatograhic separation of the flavonoids from A. moluccana. The amount of 2"-O-rhamnosylswertisin isolated (30.0 mg) was approx. twice as high as swertisin (17.5 mg). The improved surface hydrophobicity effected by the heptyl groups promoted the separation of flavonoids. From the results obtained, CH-Hp seems to be more suitable for separation of glycosylated flavonoids than other flavonoids. Thus, modified chitosan described here can be used for hydrophobic interaction chromatography as sucessfully illustrated with flavonoids. PMID- 15540597 TI - Antimicrobial activity and chemical composition of the essential oil of Nepeta crispa Willd. from Iran. AB - The composition and antimicrobial activity of the essential oil of Nepeta crispa Willd., an endemic species from Iran, was studied. The oil was obtained from the aerial parts of the plant and analyzed by GC and GC/MS. Twenty-three compounds, accounting for 99.8% of the total oil, were identified. The main constituents were 1,8-cineol (47.9%) and 4aalpha,7alpha,7abetanepetalactone (20.3%). The antimicrobial activity of essential oil of N. crispa was tested against seven gram-negative or gram-positive bacteria and four fungi. The results of the bioassays showed the interesting antimicrobial activity, in which the gram positive bacteria, Bacillus subtilis and Staphylococcus aureus, were the most sensitive to the oil. Also, the oil exhibited a remarkable antifungal activity against all the tested fungi. PMID- 15540598 TI - Antimicrobial activity of fractions and compounds from Calophyllumbrasiliense (Clusiaceae/Guttiferae). AB - Calophyllum brasiliense (Clusiaceae/Guttiferae) is a native Brazilian medicinal plant traditionally used against several diseases, including infectious pathologies. Crude methanolic extracts (CME) and two fractions, denoted non-polar (soluble in chloroform) and polar (nonsoluble in chloroform), were prepared from different parts of the plant (roots, stems, leaves, flowers and fruits) and studied. The following compounds were isolated and tested against pathogenic bacteria and yeasts by determination of the minimal inhibitory concentration (MIC): brasiliensic acid (1), gallic acid (2), epicatechin (3), protocatechuic acid (4), friedelin (5) and 1,5-dihydroxyxanthone (6). The results indicated that all the parts of the plant exhibited antimicrobial activity against Gram-positive bacteria, which are selectively inhibited by components of C. brasiliense. No activity was observed against Gram-negative bacteria and yeasts tested. Regarding the isolated compounds, substance 4 showed antimicrobial activity against all the tested microorganisms, whereas compound 6 exhibited antimicrobial activity only against Gram-positive bacteria. The results from the current study confirm and justify the popular use of this plant to treat infectious processes. PMID- 15540600 TI - The influence of growth regulators on membrane permeability in cultures of winter wheat cells. AB - The effect of plant growth substances (IAA, 2,4-D, zeatin, kinetin, zearalenone) were studied on membrane properties of the cells of embryogenic (E) and non embryogenic (NE) calli derived from immature inflorescences (inf) or embryos (emb) of winter wheat. Calli initiated from inflorescences show higher permeability. The ion leakage from cells of E calli was higher than from cells of NE calli. Growth regulators were used in concentrations of 2-30 mg/l (about 10 140 microM). All tested growth substances increased ion leakage from NE emb cells, IAA, zeatin and kinetin being most effective. In NE inf cells the effect of growth substances was similar as in NE emb, but much weaker. In E cells of both types (inf and emb) growth substances decreased ion leakage. Changes in the leakage of potassium and calcium ions were similar to those in total ion leakage. The uptake of labelled auxins (IAA and 2,4-D) was higher in NE cells (especially in NE inf) than in E cells. The endogenous level of IAA was higher in E cells than in NE cells and in inf cells than in emb cells. The importance of auxin in determining permeability of cell membranes is discussed. PMID- 15540599 TI - Biological evaluation of some selected cyclic imides: mitochondrial effects and in vitro cytotoxicity. AB - Cyclic imides such as succinimides, maleimides, glutarimides, phthalimides and their derivatives contain an imide ring and a general structure -CO-N(R)-CO- that confers hydrophobicity and neutral characteristic. A diversity of biological activities and pharmaceutical uses have been attributed to them, such as antibacterial, antifungal, antinociceptive, anticonvulsant, antitumor. In spite of these activities, much of their action mechanisms at molecular and cellular levels remain to be elucidated. We now show the effects of several related cyclic imides: maleimides (S2, S2.1, S2.2, S3), glutarimides (S4, S5, S6), 4 aminoantipyrine derivatives (L1, F1, AL1, F1.14, F1.2) and sulfonated succinimides (RO1, FA, FE, FD, MC, DMC) on isolated rat liver mitochondria, B16 F10 melanoma cell line, peritoneal macrophages and different bacterial streams. The effects on mitochondrial respiratory parameters, cell viability and antibacterial activity were also evaluated. The results indicated that S3, S5 and S6 caused an increased oxygen consumption in the presence of ADP (state III) or its absence (state IV), while all other compounds decreased those parameters at different degrees of inhibition. All the compounds decreased the respiratory control coefficient (RCC). Loss of cell viability of peritoneal macrophages and the B16-F10 cell line was observed, L1 and S2.1 being more effective. S1, S2, S3, L1 and F1 compounds showed antibacterial activity at experimental concentrations. PMID- 15540601 TI - Effect of photon flux density and temperature on the production of halogenated monoterpenes by Plocamium cartilagineum (Plocamiaceae, Rhodophyta). AB - The effect of different photon flux densities (PFD) and temperatures on the relative growth rate (RGR) and the concentration of three halogenated monoterpenes in samples of Plocamium cartilagineum L.( Dixon), a marine alga (Rhodophyceae), were studied. The highest RGR (22.8 +/- 0.04 d(-1)) was obtained at 15 degreebC and 41 ,mol m(-2) s)(-1) of PFD and the lowest (18.0 +/- 0.12 d( 1)) was obtained at 18 degrees C and 120 micromol m(-2) s(-1). The different temperatures and light used in assays did not affect significantly the production of organic compounds. The production of mertensene and violacene was not affected significantly. However, compound 1 reached the highest concentration at 15 degrees C and 65 micromol m(-2) s(-1)). The relationship between growth and production of monoterpenes of P. cartilagineum and the effect of temperature and the PFD were analyzed. PMID- 15540602 TI - Salicylic acid glucoside acts as a slow inducer of oxidative burst in tobacco suspension culture. AB - Salicylic acid beta-glucoside (SAG) is a storage form of a defense signal against pathogens, releasing free salicylic acid (SA), to meet the requirements in plants. Since excess SA induces locally restricted cell death following oxidative burst and Ca2+ influx in plants, the effects of SAG on cell viability, Ca2+ influx, and generation of superoxide (O2*-) were examined in suspension-cultured tobacco BY-2 cells expressing aequorin. Among SA-related chemicals tested, only SAG induced the slow and long-lasting O2*- generation, although SAG was less active in acute O2*- generation, Ca2+ influx and induction of cell death. The prolonging action of SAG is likely due to gradual release of SA and the data suggested that a peroxidase-dependent reaction is involved. Notably, pretreatment with low-dose SA (50 micromu) enhanced the response to SAG by 2.5-fold. There are four possible secondary messengers in early SA signaling detectable in the BY-2 culture, namely O2*-, H2O2, Ca2+ and protein kinase (PK). If these messengers are involved in the low-dose SA-dependent priming for SAG response, they should be inducible by low-dose SA. Among the four SA-inducible signaling events, PK activation was excluded from the low-dose SA action since a much higher SA dose (> 0.4 mmu) was required for PK activation. PMID- 15540603 TI - Microsomal detoxification enzymes in yam bean [Pachyrhizus erosus (L.) urban]. AB - Cytochrome P450s and glutathione-S-transferases (GSTs) constitute two of the largest groups of enzyme families that are responsible for detoxification of exogenous molecules in plants. Their activities differ from plant to plant with respect to metabolism and substrate specificity which is one of the reasons for herbicide selectivity. In the tuber forming yam bean, the legume Pachyrhizus erosus, their activities at the microsomal level were investigated to determine the detoxification status of the plant. The breakdown of the herbicide isoproturon (IPU) to two distinct metabolites, 1-OH-IPU and monodesmethyl-IPU, was demonstrated. GST activity was determined with model substrates, but also by the catalysed formation of the fluorescent glutathione bimane conjugate. This study demonstrates for the first time microsomal detoxification activity in Pachyrhizus and the fluorescence image description of microsomal GST catalysed reaction in a legume. PMID- 15540604 TI - Analysis of the labial gland secretions of the male bumblebee Bombus griseocollis (Hymenoptera: Apidae). AB - The labial gland secretions from males of the bumblebee Bombus (Separatobombus) griseocollis De Geer, a bumblebee exhibiting perching behaviour, were analysed by gas chromatography/mass spectrometry (GC/MS) in the electron impact and positive ion chemical ionization mode. The major compound of the complex mixture of alkenols, acetates, hydrocarbons, wax type esters and steroids is tetradecyl acetate, considerable amounts of hexadecyl, geranyllinaloyl, geranylgeranyl, docosyl, tetracosenyl and hexacosenyl acetate were also found. 1,3 Tetradecanediol diacetate, detected as a minor component, has not yet been identified in male bumblebee labial gland secretions. Besides small amounts of primary alcohols (tetradecanol and hexadecanol) the tertiary alcohol geranyllinalool (3,7,11,15-tetramethyl-hexadeca-1,6,10,14-tetraene-3-ol) was also present. The primary alcohols were also present as esters of butanoic, dodecanoic, tetradecanoic, and hexadecanoic acid. Besides the usual mixture of un and mono-unsaturated straight chain hydrocarbons, the labial gland contains the isoprenoid hydrocarbons beta-springene [(6E, 10E)-7,11,15-trimethyl-3-methylene hexadeca-1,6,10,14-tetraene] and two isomers of a-springene [(3Z,6E,10E)- and (3E,6E,10E)-3,7,11,15-tetramethyl-hexadeca-1,3,6,10,14-pentaene]. The close relationship in chemical composition in male bumblebees with perching and flight pass behaviour is discussed. PMID- 15540605 TI - Synthesis and field tests of sex pheromone components of the leafroller Argyrotaenia sphaleropa. AB - Female pheromone glands of the leafroller Argyrotaenia sphaleropa were analyzed. Two acetates were identified as (11Z,13)-tetradecadien-1-yl acetate and (11Z) tetradecen-1-yl acetate by comparison with synthesized references. The (11Z,13) tetradecadien-1-yl acetate and the aldehyde (11Z,13)-tetradecadienal were synthesized via a Wittig reaction. A field-trapping test showed that a lure consisting of a mixture of (11Z,13)-tetradecadienal and (11Z,13)-tetradecadien-1 yl acetate in a 10:1-ratio produced the highest trap catches. PMID- 15540606 TI - Interactions of calf spleen purine nucleoside phosphorylase with 8-azaguanine, and a bisubstrate analogue inhibitor: implications for the reaction mechanism. AB - Interactions of calf spleen purine nucleoside phosphorylase (PNP) with a non typical substrate, 8-azaguanine (8-azaG), and a bisubstrate analogue inhibitor, 9 (2-phosphonylmethoxyethyl)-8-azaguanine (PME-azaG), were investigated by means of steady-state fluorescence spectroscopy. Both 8-azaG and PME-azaG form fluorescent complexes with the enzyme, and dissociation constants are comparable to the appropriate parameters (Km or Ki) obtained from kinetic measurements. PME-azaG inhibits both the phosphorolytic and synthetic pathway of the reaction in a competitive mode. The complex of 8-azaG with PNP is much weaker than the previously reported Gua-PNP complex, and its dissociation constant increases at pH > 7, where 8-azaG exists predominantly as the monoanion (pKa approximately 6.5). The fluorescence difference spectrum of the PNP/8-azaG complex points to participation of the N(7)H or/and N(8)H tautomers of the neutral substrate, and the 9-(2-phosphonylmethoxyethyl) derivative also exists as a neutral species in the complex with PNP. The latter conclusion is based on spectral characteristics of the PNP/PME-azaG complex, confirmed by fluorimetric determination of dissociation constants, which are virtually pH-independent in the range 6-7. These findings testify to involvement of the neutral purine molecule, and not its monoanion, as the substrate in the reverse, synthetic reaction. It is proposed that, in the reverse reaction pathway, the natural purine substrate is bound to the enzyme as the neutral N(7)H tautomer, which is responsible for the reported strong fluorescence of the guanine-PNP complex. PMID- 15540607 TI - The influence of naringin on the oxidative state of rats with streptozotocin induced acute hyperglycaemia. AB - The effect of various doses (0, 10, 20, 40, or 80 mg/kg body weight) of naringin (a citrus flavonone) was studied on streptozotocin (STZ)-induced hyperglycaemic rats to evaluate the possible hypoglycaemic and antioxidant activity of naringin in diabetes. In comparison to the normoglycaemic group the treatment of rats with a single dose of STZ (65 mg/kg body weight) only revealed a significant increase (P < 0.05) in plasma hydrogen peroxide (H2O2) by 230%, increased the thiobarbituric acid reactive substances (TBARS) as index of the lipid peroxidation level by 69%, while total antioxidant activity was decreased by 36%, with a consistent significant decrease (P < 0.05) in the activity of erythrocytes antioxidative enzymes catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and paraoxonase (PON). Exogenous administration of individual gradual doses of naringin to hyperglycaemic rats causes a dose-dependent decrease of the glucose level, an increase of the insulin concentration, a decrease of the H2O2 and TBARS levels, as well as the increase of the total antioxidant status with an increase of antioxidant enzyme activities (CAT, SOD, GPx, and PON). From this study, it may be concluded that all doses of naringin provided a significant amelioration of hypoglycaemic and antioxidant activity in STZ-induced diabetic rats, however, the greatest effect of naringin was observed at 80 mg/kg body weight. PMID- 15540608 TI - A biochemical approach for detecting interactions between peptides from the HIV gp120 glycoprotein and a CD4 sequence. AB - Peptides selected from the HIV viral protein gp120 bind to a synthetic peptide mimicking sequence 78-89 of the human lymphocyte CD4 molecule, linked to activated Sepharose. The binding of viral fragments to the CD4 peptide-Sepharose beads was ascertained either by aid of a ninhydrin reagent or by fluorescence microscopy. A suitable alignment of these HIV peptides with the CD4 fragment showed that multiple interactions might occur between hydrophobic or charged groups of the two molecules. Although this experiment does not demonstrate that these two amino acid stretches are involved in the primary binding of gp120 to CD4 receptors, the present data suggest that the two sequences might have some kind of interaction during subsequent steps of viral infection. PMID- 15540609 TI - Interactions of novel morpholine and hexamethylene derivatives of anthracycline antibiotics with DNA. AB - Doxorubicin (DOX), daunorubicin (DRB), epidoxorubicin (EDOX) and their analogues with a 3'-NH2 group in daunosamine form a covalent bond with a 2-NH2 group of guanine via a methylene group from formaldehyde (CH2O). It is assumed that a Schiff base type intermediate is formed between CH2O and the 3'-NH2 group in the reaction. This reaction is supposed to occur in the cell. New analogues of anthracyclines with formamidine functionality bound to C-3' of daunosamine and containing the bulky morpholine (DRBM, DOXM and EDOXM) or hexamethyleneimine rings attached are studied in our laboratory. These substituents decrease the association of the drugs to DNA and potentially hinder the formation of Schiff base-intermediates. Our experiments indicate that the formation of the covalent complexes by DRB, DOX and EDOX under these conditions is confirmed by a high enhancement (17-40x) of the inhibition of overall RNA synthesis by E. coli RNA polymerase on T7 DNA. DRBM and DOXM exhibit a lower enhancement of the inhibition by CH2O (7-13x). The other analogues show a 1.6-3x increase of inhibition. Hence, their covalent binding is lower than that of the parent compounds. These conclusions are confirmed by spectrophotometric estimations following removal of non-covalently associated drugs. Electrophoretic analysis of drug-DNA complexes formed in the presence of CH2O indicates that DRBM and DOXM as their parent compounds induce labile cross-links in DNA. Comparison of the results obtained at the subcellular level with cytotoxicity estimations indicates that there is a correlation between cytotoxicity of the anthracyclines on L1210 cells and transcriptional template activity of drug-DNA complexes formed in the presence of CH2O (r = 0.64; n = 9). These data confirm a notion that covalent attachment of anthracyclines to DNA is an essential event leading to cytotoxicity. PMID- 15540610 TI - The ecdysteroid UDP-glucosyltransferase gene promoter from Autographa californica multicapsid nucleopolyhedrovirus. AB - The ecdysteroid UDP-glucosyltransferase (egt) gene promoter fragments of different lengths were generated from the genomic DNA of the Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) by PCR. After being purified and enzymatic digestion, they were cloned into the pGEM-3Z vector for construction of reporter plasmids pAcegt542-luc, pAcegt309-luc and pAcegt159-luc with the luciferase gene driven by the AcMNPV egt promoter. The results of transient expression in the Spodoptera frugiperda cell line-21 (Sf21) showed that the transcriptional activity of the AcMNPV egt promoter required the transactivation of viral factor(s). The expression of luciferase gene driven by the AcMNPV egt promoter was first detected at 24 h post infection. The egt promoter from the Bombyx mori nucleopolyhedrovirus (BmNPV), closely related to AcMNPV, revealed similar properties to that of the AcMNPV egt promoter. When BmNPV homologous region 3 was subcloned downstream the luciferase gene, the luciferase activity of the reporter plasmid was enhanced by over 1000-fold, but the property of the promoter was not changed. As a substrate of ecdysteroid UDP glucosyltransferase (EGT), foreign insect ecdysone showed negative effects on egt promoter transcriptional activity. Ecdysone of 1.0-2.0 microg/ml significantly down-regulated the promoter activity. Promoter activities of different lengths showed that an AcMNPV egt promoter fragment of 159 bp has the basal transcriptional activity but it was almost abolished only about 0.2% of that of 309 bp and 542 bp, respectively, and the nucleotide sequence from - 159 to - 309 nt upstream the translation initiation site includes the main cis-acting elements interacting with viral factors. PMID- 15540611 TI - Cloning and expression of Taxus acyltransferase cDNA. AB - A new full-length acyltransferase cDNA was obtained from Taxus chinensis by homology-based cloning strategy. The cDNA has an open-reading frame of 1,275 nucleotides, which encodes 425 amino acids with a calculated molecular weight of 47,241 Da and an estimated pI value of 5.93. The deduced amino acid sequence resembles the sequences of other cloned acyltransferases (56-61% identity; 71-75% similarity) involved directly in taxol biosynthetic pathways. This cDNA was expressed in Escherichia coli using the expression vector pET32a(+). The expression band corresponds to the calculated mass plus the N-terminal fusion protein derived from the vector. PMID- 15540613 TI - Florida case managers deal with the effects of back-to-back hurricanes. PMID- 15540614 TI - Reorganization merges UM and social work. PMID- 15540615 TI - Algorithm guides ICP infection investigation. PMID- 15540616 TI - Patient privacy at risk in hallways and cafeterias. PMID- 15540612 TI - Adaptation of the phosphotungstate method to determine reduced and oxidized vitamin C in blood plasma. AB - The phosphotungstate reagent (PTR) was used for quantitative spectrophotometric determination of physiological forms of vitamin C in blood plasma. An immediate action of PTR on the first half of the tested samples allowed to determine reduced vitamin C concentrations (I) at 700 nm. 10 mM dithiothreitol added to the second half of the samples reduced oxidized vitamin C in it--hence the total amount of this vitamin was reduced with a concentration (II) determined as above (remains of dithiothreitol were removed with N-ethylmaleimide). The difference of results (II) and (I) gave the concentration of oxidized vitamin C. The method is characterised by fault-less analytical parameters: correlation coefficients of analytical curves > 0.99, recovery factor 100.5%, variation coefficients intra- and inter-serial < 3% and < 5%, respectively, detection limit 0.05 microM. The simplicity of the method enables an easy control of the ratio of oxidized and reduced vitamin C concentrations in blood plasma--the biomarker of the level of oxidative damage to cells. PMID- 15540617 TI - Case managers are still fighting to prove their value. PMID- 15540618 TI - Physician-specific CM pays off for hospital. PMID- 15540619 TI - Team effort helps hospital exceed standards for CHF. PMID- 15540620 TI - Holistic wound care yields better healing rates. PMID- 15540621 TI - Diesel exhaust particles in the work environment and their analysis. AB - Diesel engines are widely used in industries, for example transportation, mining, and construction, because they efficiently produce high power. In diesel exhaust particles (DEP), the number of ultrafine particles, less than around 100 nm, is dominant in contrast to mass size distribution. Carcinogenic PAHs may be adsorbed on DEP at high concentrations. As occupational exposure usually occurs near emission sources, workers are likely to be exposed to high concentration DEP. The exhaust emissions of diesel engines have become lower by modification of the engines and fuels, and introduction of filters and catalysts, thus it has become more difficult to monitor mass and chemical components in DEP. New technology and instruments are being introduced to characterize DEP especially chemically. Recently, quick analytical methods without extraction, and continuous or semi continuous methods have been introduced. This article will review 1) Elemental Carbon (EC) monitors, 2) analytical methods of individual PAH without solvent extraction, and 3) continuous PAH monitor, because EC and PAH are typical constituents for DEP. PMID- 15540622 TI - Effects of static load on the weight and protein content in the leg muscles of the mouse: a simulation of prolonged standing in the workplace. AB - To simulate the effects of prolonged standing in the workplace on the leg muscles, we subjected mice to centrifugation for 6 wk. The absolute wet weight of leg muscles and internal organs of mice were measured after exposure to 3G by centrifugation for 6 wk and at 2 wk after removal of centrifugation. The weight of the soleus muscle (antigravity muscle) significantly increased after 6-wk exposure to centrifugation, but it decreased to its control weight 2 wk after removal of centrifugation. In contrast, the wet weights of the anterior tibial muscle, liver, and kidneys of mice centrifuged for 6 wk were significantly lower than those of the control mice; they had returned to control levels 2 wk after removal of centrifugation. It was therefore suggested that prolonged standing enlarged the leg muscles but its effect did not last for a long period of time after stopping prolonged standing. Western blot analysis of proteins extracted from the soleus muscle showed that vinculin and alpha-actinin in the centrifuged mice increased slightly, but there were no differences in the heat shock protein 70 (HSP70) and desmin levels between the centrifuged mice and control mice. No difference in HSP 70 suggested that muscle damage did not exist after 6 wk centrifugation. PMID- 15540623 TI - Exposure to urban pollutants and serum levels of IgG anti Herpes Simplex virus type 1. AB - The aim of the study is to investigate whether traffic policemen exposed to urban pollutants and psycho-social stressors may be at risk of modifications in serum levels of immunoglobulins G antibodies (IgG Ab) against Herpes Simplex Virus (type1) (HSV-1) compared with controls. Traffic policemen were matched by sex, age, working life and drinking habits (less than two glasses of wine or beer per day) with controls, after excluding the subjects with the principal confounding factors (smoking habit, use of paints, solvents, pesticides and drinking habits). Were included in the study 125 traffic policemen (54 men and 71 women) with outdoor activity and 125 controls (54 men and 71 women) with indoor activity. Mean levels of IgG Ab against HSV-1 were significantly higher in traffic policemen of both sexes compared with controls. The frequency of workers with IgG Ab against HSV-1 levels higher than our normal laboratory values, was significant in traffic policemen compared with controls of both sexes. The distributions of IgG Ab against HSV-1 levels in traffic policemen compared with controls of both sexes were significant. The authors hypothesise an effect of the working activity in traffic policemen on serum levels of IgG Ab against HSV-1 compared to controls. PMID- 15540624 TI - Lead levels in urine of never-smoking adult women in non-polluted areas in Japan, with references to cadmium levels in urine. AB - The present study was initiated to assess current level of general population exposure to lead (Pb) in terms of Pb in urine (Pb-U). For this purpose, spot urine samples were collected from 2,332 never-smoking adult women in 10 areas all over Japan, and were subjected to analysis for Pb-U by graphite furnace atomic absorption (GFAA). Data on cadmium in urine (Cd-U) as measured by GFAA were cited from a previous publication). The grand geometric means (GM) for Pb-U as observed, as corrected for creatinine (cr) and as corrected for a specific gravity of 1.016 were 1.06 microg/l, 1.28 microg/g cr and 1.14 microg/l, respectively, with small intra- and inter-variations with geometric standard deviations of about 2 or less. Arithmetic means of the 10 GMs (one each for the 10 areas) were 1.05 microg/l, 1.19 microg/g cr and 1.18 microg/l, respectively. The levels observed appeared to be lower than levels reported for other populations in the world. Chronological comparison within Japan suggested a gradual decrease in Pb-U in past over 20 yr. No close correlation was detected between Pb-U and Cd-U. PMID- 15540625 TI - Fatal occupational injuries in the construction sector in Kocaeli, Turkey, 1990- 2001. AB - In Turkey, the construction sector is responsible for the largest number of work related fatalities among all industries. This study's objective is to determine the characteristics of the fatalities in the construction sector in Kocaeli Province. The study was carried out retrospectively by investigating the records of occupational deaths between 1990 and 2001 in the Kocaeli District of Turkey. Fatal occupational injuries in the construction sector were investigated in detail by evaluating the records. The analysis was based on data from 153 deaths. The incidence of occupational injuries was found overall to be 4.5% in Kocaeli, while the annual mortality rate was 60.4 in Kocaeli and 79.0 in Turkey per 100,000 workers over the entire time period. On the other hand, the fatality rate of occupational injuries was found to be 12.0 per 1,000 in Turkey and 6.4 per 1,000 in Kocaeli. The evaluation of the type of injuries in construction sector has revealed that in 69 of cases (45.1%) fell down from high altitude, in 22 cases (14.4%) vehicle accident, and in 22 (14.4%) electrocution were the cause of death. The leading causes of death have been found to be blunt-force trauma (37.9%) and head trauma (25.5%). Fourty one percent of those who had occupational accidents have had less than one year work experience. Only in 7.8% of deaths, the cause was determined through an autopsy. PMID- 15540626 TI - A study on the effect of lead on event-related potentials among lead-exposed workers. AB - OBJECTIVES: In order to clarify the effect of lead on higher cerebral functions, lead-exposed workers (Pb group) and controls were examined for event-related potentials. SUBJECTS AND METHODS: Fourteen lead-exposed workers with a mean age of 57.1 yr (SD=4.27, range 48-64; lead concentration of whole blood ranged from 33 to 106 microg/dl with a mean of 58.6 and SD 28.6 microg/dl) and 19 age-matched control workers with a mean age of 57.3 yr (SD=4.80, range 48-65) were examined. Visual P300 was recorded by button pushing to the target image (minute checkerboard pattern, 20%), and the NO-GO potential by no button pushing to the target image (same as above, 50%). RESULTS: Latencies of P300 in the Pb group (475+/-46.0 ms) were significantly delayed compared with those in controls (407+/ 42.4 ms, p<0.01 by Student's t test). Amplitudes of the NO-GO potential in the Pb group (4.59+/-2.04 microV) significantly increased compared with those in the controls (3.18+/-1.41 microV, p<0.05). CONCLUSION: The finding suggests that lead exposure affects high cerebral functions of cognition and attention, but is unclear in suppression of movement. PMID- 15540627 TI - Malignant pleural mesothelioma in parts of Japan in relationship to asbestos exposure. AB - Malignant pleural mesothelioma is induced by asbestos exposure. Many reports have described this situation in America and European countries, but a few have been published in Japan. In this study malignant pleural mesothelioma cases in hospitals located in an area facing the Seto Inland Sea were evaluated. A total of 106 patients were examined with 100 patients having had occupational exposure to asbestos and 6 patients without such histories of asbestos exposure. Ninety seven were male and 9 were female. Ages ranged from 41 to 87 yr with mean of 64.8+/-5.3 yr. Thirty seven cases showed epithelial type of tumor, 25 biphasic type and 15 showed sarcomatous. The remaining 23 cases had insufficient evidence for typing the tumor. The mean survival rate for all cases was 9.2+/-11.6 months. Fifty-one patients had occupational histories of shipyard work, 16 patients worked in asbestos cement piping, and the remainder were employed in miscellaneous jobs related asbestos exposure. The duration of asbestos exposure ranged up to 20 yr or longer with the mean of 17.2+/-8.9 yr and the average latent period for the occurrence of malignant pleural mesothelioma was more than 31 yr with the mean of 37.0+/-13.3 yr. Quantification of asbestos bodies in the lungs indicated a high concentration in most patients and the major types of asbestos fibers were crocidolite and amosite. Six cases appeared after exposure to chrysotile. These results indicated that ninety four percent of malignant pleural mesothelioma appeared due to the exposure to asbestos including crocidolite and amosite. The remainder may be blamed on exposure to chrysotile. PMID- 15540628 TI - Occupational health education and collaboration for reducing the risk of lead poisoning of workers in a battery manufacturing plant in Thailand. AB - We provided occupational health education as training to all workers (N=31) and managers, and collaborated with them against reducing the risk of lead poisoning of workers at an assembly section in a battery manufacturing plant in Bangkok, Thailand in 2002. After occupational health education, many workers (80.6-100.0%) noticed and understood the toxicity of lead and the importance of protection against it. Many workers regularly wore long sleeved shirt and trousers, and used the appropriate mask all day long. They changed the attitude toward their work and improved personal hygiene, for example no smoking in the workplace, washing their hands by a detergent before drinking water or having lunch, and taking a bath after the work. They especially took off working clothes at an office outside the workplace and washed them everyday. The average blood lead level of the workers significantly (P=0.002) reduced from 32.7 microg/dl to 22.4 microg/dl, although airborne lead level in the workplace remained unchanged with before conditions. From these results occupational health education and collaboration between workers and managers were effective tools to reduce the risk of lead poisoning of workers in a battery manufacturing plant. PMID- 15540629 TI - Health effects of exposure to ethylene glycol monoethyl ether in female workers. AB - Ethylene glycol monoethyl ether (EGEE) is a solvent commonly used in industry. To find the health effect of the solvent exposure in women, we did an investigation on 32 female workers exposed to EGEE in factories manufacturing photopolymer sensitization plate, and 20 subjects working in the same companies without potential exposure to the solvent. The mean age was 35.0 and 33.9 yr in the two groups, respectively. The mean concentration of the urinary metabolite (ethoxyacetic acid) was 120.87 mg/g creatinine (geometric mean) in the exposed group, and 2.71 mg/ g creatinine in the control group. Average RBC count and hemoglobin levels were normal in both groups. However, there were 2 subjects in the exposed group with an RBC count and hemoglobin concentration slightly lower than the standard. Out of 20 controls, 5 subjects reported irregular menstruation, and in comparison, 4 out of 32 exposed females had the same complaint. The most common health complaints were dizziness and swelling of the legs, with the same frequencies seen in both groups. Overall, our study suggests that although female workers were exposed to high concentrations of EGEE, subsequent health problems possibly due to such exposure were not significant. PMID- 15540630 TI - Achieving implant success through prosthetic design. Interview. PMID- 15540631 TI - Evidence based medicine. PMID- 15540632 TI - Neuropsychology of academic and behavioural limitations in school-age survivors of bacterial meningitis. AB - Neuropsychological impairments possibly underlying academic and/or behavioural limitations were studied in 149 school-age survivors of bacterial meningitis, 68 with and 81 without academic and/or behavioural limitations. Academic limitations affected mathematics, reading, and writing. Behavioural limitations were inferred from scores in the clinical range on the Child Behaviour Checklist. These children had been selected from a cohort of 674 children (57% males) who had recovered from non-Haemophilus influenzae type B bacterial meningitis and who had a mean age at infection of 2 years 4 months (range 1mo to 9y 5mo). They had neither 'complex onset' meningitis, prior cognitive or behavioural problems, nor severe disease sequelae. They were assessed with standardized assessment methods a mean of 7.8 years (range 4 to 10.4) after meningitis. Children with limitations (32% of the cohort) performed generically poorly on measures of cognitive functioning, speed, and motor steadiness, rather than having impairments in specific neuropsychological domains. The presence of two or more minor neurological signs was more frequent in the group with than in the group without limitations (30% versus 9%); this may explain the relatively poor speed and motor steadiness of the group with limitations. PMID- 15540633 TI - Correlative value of magnetic resonance imaging for neurodevelopmental outcome in periventricular leukomalacia. AB - The aim of this study was to evaluate the correlative value of magnetic resonance imaging (MRI) in children with periventricular leukomalacia (PVL) for neurodevelopmental outcome. MRI examinations of 89 children (46 males, 43 females) with PVL (median age 4y, range 1 to 14y) were reevaluated. PVL was graded as follows: grade I, unilateral or bilateral areas of periventricular hyperintensity (1-3); grade II, hyperintensity more than 3; grade III, hyperintense lesions more than 3 and ventricular wall irregularity; grade IV, diffuse PVL and ventricular dilatation. Localizations of PVL and brain abnormalities associated with PVL were also noted. Assignment to PVL grades on MRI was as follows: PVL I (n=22), PVL II (n=18), PVL III (n=30), and PVL IV (n=19). Cerebral palsy was slightly less common in children with PVL I and II compared with PVL III to IV. Motor function was normal in 50% of children with PVL grade I, but severely impaired in 73.7% of children with PVL grade IV. Results of visual function were normal in all with PVL I, but pathological in 42.1% of patients with PVL IV. Developmental tests were appropriate for age in 75% of patients with PVL I, but significantly delayed in all patients with PVL IV. Thinning of the corpus callosum and presence of cortical atrophy were also correlated with neurological outcome. Significant risk factors associated with developmental delay were asphyxia at birth (odds ratio [OR] 4.3), PVL localization numbers over 3 (OR 4.4), PVL III to IV (OR 15), thinning of corpus callosum, and cortical atrophy. PMID- 15540634 TI - Effect of intensive neurodevelopmental treatment in gross motor function of children with cerebral palsy. AB - This study examined the effect of neurodevelopmental treatment (NDT) and differences in its intensity on gross motor function of children with cerebral palsy (CP). Participants were 34 children (12 females, 22 males; mean age 7y 3mo [SD 3y 6mo], age range 3 to 14y) with mild to moderate spasticity and hemiplegia (n=10), diplegia (n=12), and tetraplegia (n=12). Gross Motor Function Classification System levels were: I (n=10), II (n=10), and III (n=14). The paired sample, which was obtained by ratio stratification and matching by sex, age, and distribution of impairment from a total of 114 children with CP, was assigned randomly to two groups: group A underwent NDT twice a week and group B five times a week for 16 weeks. The outcome measure used was the Gross Motor Function Measure, which assessed the performance of the children before and after intervention. The paired-sample t-test revealed that gross motor function of children from both groups improved significantly after intervention (p<0.05). Children in group B performed better and showed significantly greater improvement than those in group A (p<0.05). Results support the effectiveness of NDT and underline the need for intensive application of the treatment. PMID- 15540635 TI - Bimanual coordination during a goal-directed task in children with hemiplegic cerebral palsy. AB - Most studies of impaired hand function in children with hemiplegic cerebral palsy (CP) have focused on either the involved or the non-involved extremity in isolation. Coordination of the involved and non-involved hand during bimanual tasks in these children is not well understood. The present study examined bimanual coordination using a drawer-opening task under speed and hand constraints in 10 children with hemiplegic CP (5 males and 5 females, mean age 13y 5mo, range 8y to 16y) and 10 age-matched right-handed developing typically children (6 males and 4 females, mean age 13y 1mo). Children were asked to reach forward and open a drawer with one hand and then activate a light switch inside the drawer with the contralateral hand. The role of the two hands (open drawer and activate switch) and speed (self-paced vs fast-as-possible) were varied. The children with hemiplegic CP were slower (p<0.001) and less coordinated in this task, with reduced movement overlap of the two hands (p<0.001) and sequential completion of the two movement objectives (p<0.001). Moreover, the hand used for each task subcomponent affected task performance for the children with hemiplegic CP (p<0.05). Interestingly, faster speed facilitated better bimanual coordination for the children with hemiplegic CP (p<0.001). Results highlight the importance of movement constraints on task performance and suggest that movement speed might facilitate better bimanual coordination. PMID- 15540636 TI - Internal representation of movement in children with developmental coordination disorder: a mental rotation task. AB - Recent studies show that children with developmental coordination disorder (DCD) have difficulties in generating an accurate visuospatial representation of an intended action, which are shown by deficits in motor imagery. This study sought to test this hypothesis further using a mental rotation paradigm. It was predicted that children with DCD would not conform to the typical pattern of responding when required to imagine movement of their limbs. Participants included 16 children with DCD and 18 control children; mean age for the DCD group was 10 years 4 months, and for controls 10 years. The task required children to judge the handedness of single-hand images that were presented at angles between 0 degrees and 180 degrees at 45 degrees intervals in either direction. Results were broadly consistent with the hypothesis above. Responses of the control children conformed to the typical pattern of mental rotation: a moderate trade off between response time and angle of rotation. The response pattern for the DCD group was less typical, with a small trade-off function. Response accuracy did not differ between groups. It was suggested that children with DCD, unlike controls, do not automatically enlist motor imagery when performing mental rotation, but rely on an alternative object-based strategy that preserves speed and accuracy. This occurs because these children manifest a reduced ability to make imagined transformations from an egocentric or first-person perspective. PMID- 15540637 TI - Voxel-based morphometry elucidates structural neuroanatomy of high-functioning autism and Asperger syndrome. AB - Efforts to examine the structural neuroanatomy of autism by using traditional methods of imaging analysis have led to variable findings, often based on methodological differences in image acquisition and analysis. A voxel-based computational method of whole-brain anatomy allows examination of small patterns of tissue differences between groups. High-resolution structural magnetic resonance images were acquired for nine males with high-functioning autism (HFA; mean age 14y [SD3y 4mo]), 11 with Asperger syndrome (ASP; mean age 13y 6mo [SD2y 5mo]), and 13 comparison (COM) participants (mean age 13y 7mo [SD 3y 1mo]). Using statistical parametric mapping, we examined contrasts of gray matter differences between the groups. Males with HFA and ASP had a pattern of decreased gray matter density in the ventromedial regions of the temporal cortex in comparison with males from an age-matched comparison group. Examining contrasts revealed that the COM group had increased gray matter density compared with the ASP or combined HFA and ASP group in the right inferior temporal gyrus, entorhinal cortex, and rostral fusiform gyrus. The ASP group had less gray matter density in the body of the cingulate gyrus in comparison with either the COM or HFA group. The findings of decreased gray matter density in ventromedial aspects of the temporal cortex in individuals with HFA and ASP lends support to theories suggesting an involvement of these areas in the pathophysiology of autism, particularly in the integration of visual stimuli and affective information. PMID- 15540638 TI - Periodic leg movements in prepubertal children with sleep disturbance. AB - This study's aims were to determine: (1) prevalence of periodic leg movements (PLMs) in walking prepubertal children consulting a sleep clinic for any sleep disorder; (2) associations between PLMs and other sleep and medical disorders; and (3) the response of other sleep disorders to treatment with the dopamine agonist pramipexol. Clinical evaluation and polysomnography were carried out for a period of 12 months on 252 consecutively seen, prepubertal children with sleep disorders (156 males, 96 females; aged 15mo to 11y, mean 7y 1mo, SD3y 10mo). Sleep disorders unrelated to PLMs were treated, and six children received pramipexol for PLMs. Follow-up included clinical evaluation and polysomnography. Twenty-three per cent of children were diagnosed with PLMs on the basis of polysomnography. The presence of PLMs had usually been unrecognized clinically. The only clinical symptom that could be related to periodic limb movement disorder was a report of leg pains at morning awakening. Only two of 58 children had PLMs without other clinical or polysomnographic findings. Comorbidity seen with PLMs included neuropsychiatric syndromes (n=20), isolated sleep disordered breathing (SDB; n=29), and several other comorbid conditions (n=7). Seven of 11 children seen with attention-deficit-hyperactivity disorder also had PLMs. Surgery for SDB was associated with subsequent cessation of PLMs in 15 of 29 children. Five out of six children with PLMs who received pramipexol were able to tolerate the drug and experienced a complete disappearance of their PLMs. Presence of chronic fatigue, sleepiness, disrupted nocturnal sleep, and difficulties in falling asleep should lead to a systematic search for PLMs that is independent of associated syndromes. Isolated treatment of SDB might help eliminate some, but not all, PLMs. PMID- 15540640 TI - Melatonin therapy for circadian rhythm sleep disorders in children with multiple disabilities: what have we learned in the last decade? PMID- 15540639 TI - Systematic review of melatonin treatment in children with neurodevelopmental disabilities and sleep impairment. AB - Sleep disturbances in children with neurodevelopmental disabilities are common and frequently difficult to treat with conventional pharmacological and behavioural methods. Melatonin is a pineal hormone known to be important in the regulation of the circadian rhythm, including the sleep-wake cycle. This systematic review of available evidence from randomized clinical trials assesses whether melatonin plays a beneficial role in these children and, in particular, its effect on total sleep time, time to sleep onset (sleep latency), and number of awakenings. We also looked at a parental view of the effect. Randomized clinical trials were identified where oral melatonin was compared with a placebo in children with any type of neurodevelopmental disability and associated sleep disturbance. Only three studies, reporting a total of 35 children, fulfilled the criteria for inclusion. The two studies that reported time to sleep onset showed a significant decrease (p<0.05) in this specific outcome where melatonin was compared with a placebo. There was no significant effect of melatonin compared with a placebo on the other outcome measures of total sleep time, night-time awakenings, and parental opinions. Despite the extremely limited randomized clinical trial data, melatonin appears to remain a commonly prescribed drug for disturbed sleep in children with neurodevelopmental abnormalities. PMID- 15540641 TI - Sturge-Weber syndrome and paroxysmal hemiparesis: epilepsy or ischaemia? AB - Transient neurological deficits experienced by patients with Sturge-Weber syndrome can be caused by epilepsy, or may result from temporary ischaemia of the cortex underlying the vascular malformation. To show the difficulty in distinguishing seizures from ischaemic symptoms, two male children with episodes of acute unilateral weakness are presented here as well as a review of the literature. The first child presented at 2 years of age with a sudden increase in his pre-existing right hemiparesis accompanied by screaming. Ictal epileptiform activity was recorded at the moment of the attack, and subsequent seizures were controlled by adjustment of antiepileptic drug treatment. The second child presented at 4 years of age with attacks of vomiting and a coinciding increase in the pre-existing paresis of the left leg. Electroencephalogram (EEG) recording did not show ictal epileptiform activity. The origin was presumed to be vascular. Treatment with aspirin led to control of these transient ischaemic attacks. Ictal EEG is needed to differentiate between an epileptic and an ischaemic origin of transient focal deficit. Treatment with aspirin should be considered if an ischaemic origin cannot be excluded. PMID- 15540642 TI - 'Effects of oral baclofen on children with cerebral palsy'. PMID- 15540643 TI - Weight and height gain after intrathecal baclofen pump implantation in children with spastic tetraparesis. PMID- 15540644 TI - 'A child with severe night terrors and sleep-walking responds to melatonin therapy'. PMID- 15540645 TI - Milk powders ageing: effect on physical and functional properties. AB - Milk powders are now considered as food ingredients, mainly because of the functional properties of milk proteins. During the storage of milk powders, many physicochemical damages, mainly dependent on lactose glass transition occur. They have important consequences on physical (flowability) and functional properties (solubility, emulsifying, and foaming properties) of milk powders. First, lactose crystallization modifies the microstructure and chemical composition of the surface of powder particles. Thus, milk powders flowability is decreased. Since the structure of milk proteins is destabilized, its solubility is damaged. Moreover, particle collapse and caking occur and mainly decrease the physical properties of milk powders (density and flowability). The mechanical stresses involved may also enhance proteins unfolding, which is detrimental to solubility. Finally, molecular mobility is favored upon ageing, and both chemical (Maillard reaction) and enzymatic reactions occur. Maillard reaction and oxidation enhance protein interactions and aggregations, which mainly lessen milk powders solubility. Maillard reaction also decreases emulsifying and foaming properties. Storage temperature and relative humidity have been considered as the predominant factors involved, but time, milk components, and their physical state also have been implied. PMID- 15540646 TI - A review of acrylamide: an industry perspective on research, analysis, formation, and control. AB - Acrylamide is a synthetic monomer with a wide scope of industrial applications, mainly as a precursor in the production of several polymers, such as polyacrylamide. The main uses of polyacrylamides are in water and wastewater treatment processes, pulp and paper processing, and mining and mineral processing. The announcement by the Swedish National Food Administration in April 2002 of the presence of acrylamide predominantly in heat-treated carbohydrate rich foods sparked intensive investigations into acrylamide, encompassing the occurrence, chemistry, agricultural practices, and toxicology, in order to establish if there is a potential risk to human health from the presence of this contaminant in the human diet. The link of acrylamide in foods to the Maillard reaction and, in particular, to the amino acid asparagine has been a major step forward in elucidating the first feasible chemical route of formation during the preparation and processing of food. Other probably minor pathways have also been proposed, including acrolein and acrylic acid. This review addresses the analytical and mechanistic aspects of the acrylamide issue and summarizes the progress made to date by the European food industries in these key areas. Essentially, it presents experimental results generated under laboratory model conditions, as well as under actual food processing conditions covering different food categories, such as potatoes, biscuits, cereals, and coffee. Since acrylamide formation is closely linked to food composition, factors such as the presence of sugars and availability of free amino acids are also considered. Many new findings that contribute towards a better understanding of the formation and presence of acrylamide in foods are presented. Many national authorities across the world are assessing the dietary exposure of consumers to acrylamide, and scientific projects have commenced to gather new information about the toxicology of acrylamide. These are expected to provide new scientific knowledge that will help to clarify whether or not there is a risk to human health from the consumption of foods containing low amounts of acrylamide. PMID- 15540647 TI - A new quality parameter in tomato and tomato products: ergosterol. AB - The poor precision of the "percentage of discarded fruits" and "Howard mold count" methods has increased the importance of ergosterol for the microbiological quality evaluation of tomato and tomato products. Ergosterol, a constituent of the cell wall of some important vegetable parasites, such as molds, has been recently recognized as a potential objective parameter useful for the characterization of the quality of processing tomatoes. Thus, 15 mg of ergosterol/kg total solids has been stated as an acceptability maximum limit of tomato products. PMID- 15540648 TI - Target selection in designing pasteurization processes for shelf-stable high-acid fruit products. AB - This study is focused on the search for targets and criteria for the design of pasteurization processes for high-acid shelf-stable fruit products, such as juices, nectars, pastes, purees, concentrates, jams, jellies, etc. First, an overview of pasteurization is presented and then, frequently used targets for pasteurization processes are reviewed Enzymes naturally present in fruits, in decreasing order of heat resistance, were pectinesterase, peroxidase, and polyphenoloxidase, and they may be used as pasteurization targets. The heat resistance of each enzyme is strongly dependent on its fruit origin. The most heat resistant micro-organisms capable of spoiling high acid fruit products include ascospores of Neosartorya fischeri, Byssochlamys nivea, Talaromyces flavus, Eupenicillium javanicum, and Byssochlamys fulva moulds, as well as bacterial spores of Clostridium butyricum, Bacillus coagulans, and Bacillus megaterium. These micro-organisms, spores, and enzymes were, in general, less heat resistant than the spores of a particular spoilage micro-organism named Alicyclobacillus acidoterrestris, which has been causing problems in the fruit industry. Therefore, the use of Alicyclobacillus acidoterrestris spores as a reference micro-organism in the design of pasteurization processes for high-acid shelf-stable fruit products is suggested. PMID- 15540649 TI - Health benefits of soy isoflavonoids and strategies for enhancement: a review. AB - Soybean consumption has been linked to a reduced risk for certain cancers and diseases of old age. The health benefits associated with soybean consumption have been linked to the action of isoflavonoids, the major phenolic phytochemicals found in soybean. Isoflavonoids possess numerous biological activities that may support chemoprevention through the promotion of apoptosis in diseased cells. In this study, we discuss the current state of knowledge concerning soybean isoflavonoids, their chemopreventive actions against postmenopausal health problems, cancer, and cardiovascular disease, and also biotechnology approaches toward the enrichment of soybean for isoflavonoid content. PMID- 15540650 TI - Promises and problems of functional foods. AB - "Functional" foods are branded foods, which claim, explicitly or implicitly, to improve health or well being. We review typical functional foods and their ingredients, efficacy, and safety. We also review regulations for health claims for foods worldwide. These regulations often allow manufacturers to imply that a food promotes health without providing proper scientific evidence. At the same time, regulations may ban claims that a food prevents disease, even when it does. We offer a plea for regulations that will permit all health claims that are supported by the totality of scientific evidence, and ban all claims that suggest an unproven benefit. PMID- 15540652 TI - [Study on vascular proliferation during the growth of an adrenal autograft in a rat]. PMID- 15540651 TI - Structural, biological, and evolutionary relationships of plant food allergens sensitizing via the gastrointestinal tract. AB - The recently completed genome sequence of the model plant species Arabidopsis has been estimated to encode over 25,000 proteins, which, on the basis of their function, can be classified into structural and metabolic (the vast majority of plant proteins), protective proteins, which defend a plant against invasion by pathogens or feeding by pests, and storage proteins, which proved a nutrient store to support germination in seeds. It is now clear that almost all plant food allergens are either protective or storage proteins. It is also becoming evident that those proteins that trigger the development of an allergic response through the gastrointestinal tract belong primarily to two large protein superfamilies: (1) The cereal prolamin superfamily, comprising three major groups of plant food allergens, the 2S albumins, lipid transfer proteins, and cereal alpha amylase/trypsin inhibitors, which have related structures, and are stable to thermal processing and proteolysis. They include major allergens from Brazil nut, peanuts, fruits, such as peaches, and cereals, such as rice and wheat; (2) The cupin superfamily, comprising the major globulin storage proteins from a number of plant species. The globulins have been found to be allergens in plant foods, such as peanuts, soya bean, and walnut; (3) The cyteine protease C1 family, comprising the papain-like proteases from microbes, plants, and animals. This family contains two notable allergens that sensitize via the GI tract, namely actinidin from kiwi fruit and the soybean allergen, Gly m Bd 30k/P34. This study describes the properties, structures, and evolutionary relationships of these protein families, the allergens that belong to them, and discusses them in relation to the role protein structure may play in determining protein allergenicity. PMID- 15540653 TI - [Prediction of perineural infiltration of biliary cancer by using the genes specially selected from the cDNA array data]. PMID- 15540654 TI - [Analysis of immune controlling CD4+CD25+ T cells in HTLV-1 transgenic rats]. PMID- 15540655 TI - [Study on changes in physiological [18F]fluorodeoxyglucose accumulation in the digestive tube caused by drug treatment]. PMID- 15540657 TI - [Can myocardial Na+ channel gene mutation be a prognostic factor for sudden death in asymptomatic Brugada syndrome?]. PMID- 15540656 TI - [Suppression of tissue factor expression by the control of macrophage migration inhibitory factor]. PMID- 15540658 TI - [Analysis of 4q LOH in hepatoid adenocarcinoma, common gastric cancer, and hepatocellular carcinoma]. PMID- 15540659 TI - [Analysis of the mechanism involved in the neuroendocrine-related gene expression in a small cell lung carcinoma cell strain by correlation profiling of cDNA array data]. PMID- 15540660 TI - [Enhancement of oncogene expression in myocardial cells caused by applying hydrostatic pressure]. PMID- 15540661 TI - [Study on 17q25 triplet repeat elongation and an expression enhancement phenomenon in familial moya-moya families--detection of the genes responsible for familial moya-moya disease]. PMID- 15540662 TI - [Evaluation of HTLV-1 virus expression by introducing human CRM1 gene into T cells of rats with a sustained HTLV-1 infection]. PMID- 15540663 TI - [Expression of GDNF (glial cell line-derived neurotrophic factor) and Ret in normal human and cancerous pancreatic tissues]. PMID- 15540664 TI - [Study on the mechanism to maintain calcium mobilization in platelets of patients with bipolar affective disorders--the relationship with the protein kinase C system]. PMID- 15540665 TI - A palliative care association for the whole of Africa. PMID- 15540666 TI - Place of death: hospital-based advanced home care versus conventional care. A prospective study in palliative cancer care. AB - The purpose of this prospective nonrandomized study was to evaluate time spent at home, place of death and differences in sociodemographic and medical characteristics of patients, with cancer in palliative stage, receiving either hospital-based advanced home care (AHC), including 24-hour service by a multidisciplinary palliative care team or conventional hospital care (CC). Recruitment to the AHC group and to the study was a two-step procedure. The patients were assigned to either hospital-based AHC or CC according to their preferences. Following this, the patients were asked to participate in the study. Patients were eligible for the study if they had malignant disease, were older than 18 years and had a survival expectancy of 2-12 months. A total of 297 patients entered the study and 280 died during the study period of two and a half years, 117 in the AHC group and 163 in the CC group. Significantly more patients died at home in the AHC group (45%) compared with the CC group (10%). Preference for and referral to hospital-based AHC were not related to sociodemographic or medical characteristics. However, death at home was associated with living together with someone. Advanced hospital-based home care targeting seriously ill cancer patients with a wish to remain at home enable a substantial number of patients to die in the place they desire. PMID- 15540667 TI - Listen to the patient: quality of life of patients with recently diagnosed malignant cord compression in relation to their disability. AB - Quality of life (QOL) was determined using the shortened version of the Schedule for the Evaluation of Individual Quality of Life (SEIQoL-Dw), in 180 patients, shortly after a diagnosis of malignant cord compression. The median SEIQoL score was 66 (maximum 100). Twenty-five per cent of patients had a score >83. SEIQoL correlated with KPS (P<0.004) but the range of scores for each level of performance was very wide. Although there was an association between SEIQoL and ability to walk, the difference was small and not significant. SEIQoL was adversely affected by emotional distress as measured using HAD (P<0.01), although few patients were identified as being severely depressed. The findings are a reminder that QOL as determined by the patient may not coincide with health professionals' opinions. PMID- 15540668 TI - Noninvasive mechanical ventilation as a palliative treatment of acute respiratory failure in patients with end-stage solid cancer. AB - Noninvasive ventilation (NIV) is widely used in the treatment of acute respiratory failure (ARF), but not in patients with end-stage solid cancer in whom any form of mechanical ventilation tends to be avoided. In a prospective study, we investigated the use of NIV in 23 patients with solid malignancies receiving palliative care and who were affected by severe hypoxic or hypercapnic ARF. The most frequent causes of ARF were exacerbations of pre-existing pulmonary diseases and pneumonia. After one hour, NIV significantly improved PaO2/FiO2 (from 154+/-48 to 187+/-55) and the Borg dyspnoea score (from 5.5+/-1.2 to 2.3+/ 0.3). NIV also improved pH, but only in the subset of hypercapnic patients. Thirteen of 23 (57%) patients were successfully ventilated and discharged alive, while 10/23 patients (43%) met the criteria for intubation or died after an initial trial of NIV. Only two of these patients accepted invasive ventilation. The mortality rate in this subgroup was 9/10 (90%). A higher Simplified Acute Physiology Score (SAPS II) and a lower PaO2/FiO2 on admission were associated with a lower probability of survival. Patients with ARF and end-stage solid malignancies have an overall ICU and 1-year mortality rate of 39% and 87%, respectively, but despite this, a consistent subset of patients may still be successfully treated with NIV, if the cause of ARF is reversible. PMID- 15540669 TI - Meanings of being old and living with chronic obstructive pulmonary disease. AB - The aim of this study was to achieve a deeper understanding of the meaning of the lived experiences of elderly persons who are severely ill with chronic obstructive pulmonary disease (COPD) and in need of everyday care. Narratives from dialogues with six persons were interpreted using phenomenological hermeneutic methodology. The structural analyses showed severe problems in daily life due to old age with a decaying body and dialectic experiences of resignation contentedness, loneliness-connectedness, and being homeless-being at home. The interpreted whole reveals that life, suffering and comfort must be understood as an interlaced experience where ageing is just as important as the experience of illness. These elderly people's concerns and problems due to old age, a decaying body and being severely ill with COPD call for palliative and comfort care and thus challenge all professionals involved in their care. PMID- 15540670 TI - Living with severe chronic obstructive pulmonary disease (COPD): perceptions of patients and their carers. An interpretative phenomenological analysis. AB - BACKGROUND: A study designed to explore the experiences of patients with severe chronic obstructive pulmonary disease (COPD) and their carers, particularly with regard to ongoing and palliative care needs. METHODS: The participants were nine men and one woman with severe COPD and the carers of eight of the men, in East Devon, UK. Semi-structured interviews were undertaken, transcribed and analysed using interpretative phenomenological analysis (IPA). RESULTS: The emergent themes were of losses, adaptation, relationships with health professionals and effect on carer. Losses reflected the loss of personal liberty and dignity and of previous expectations of the future. Adaptation included strategies to cope with the effects of the disease. Relationships related to both positive and negative aspects of contact with health professionals. There was appreciation for continuity of care and reassurance. The effect on the carer was evident particularly as they had to take on multiple roles. They also experienced some of the same losses as the patient and appeared enmeshed with the illness. CONCLUSIONS: This study confirmed the inexorable decline in activities of daily life and social isolation for patients with severe COPD. Adaptive strategies were common and some positive aspects were identified. Support from the primary health care team was appreciated. The strain on carers was very apparent. The concept of a more structured sharing of information and a surveillance role mediated by health care professionals known to the patient and carer would be a pragmatic approach to improving care. PMID- 15540671 TI - Attitudes of 100 patients with chronic obstructive pulmonary disease to artificial ventilation and cardiopulmonary resuscitation. AB - One hundred clinically stable outpatients with chronic obstructive pulmonary disease were surveyed at home by respiratory nurse specialists (RNS) about their views towards cardiopulmonary resuscitation (CPR) and in hospital ventilation. Written information about COPD, CPR and ventilation was provided and consent obtained. The breathing problem-based quality-of-life questionnaire (BP-QoL) was completed. The following information was recorded: age, sex, spirometry, hospital admissions, and antidepressant and oxygen usage in the previous year. Patients were then asked to imagine a scenario in which they were admitted to hospital and their chest condition deteriorated in spite of standard treatment. 'Having reached that stage would you wish to have noninvasive ventilation (NIV), invasive ventilation (IV) or CPR?' Three months later patients were asked to complete a postal patient satisfaction questionnaire. RESULTS: Of 100 patients 41 were male and the mean age was 74.1 years. Fifty four patients had a FEV1 <40% and 37 had a FEV1 between 40 and 59% predicted. Twenty-four patients were on long-term oxygen therapy, eight had taken antidepressants and 56 had been admitted to hospital in the previous year. Forty-eight patients wanted all additional treatments to be attempted if needed and 12 wanted none. Nineteen patients said 'no' for CPR and 10 said 'no' for CPR and IV. There was no significant statistical difference between the groups answering 'yes' or 'no'. Seventy-six per cent of patient satisfaction questionnaires were returned. All patients were satisfied with the way they had been approached and the information received, and 98% of them thought that this issue should be discussed with all patients. CONCLUSION: Attitudes towards resuscitation can be discussed with COPD patients by RNS without causing distress. Attitudes to resuscitation could not be predicted from parameters of respiratory disease severity or age. PMID- 15540672 TI - Sexuality in palliative care: patient perspectives. AB - This qualitative study investigated the meaning of sexuality to palliative patients. Face-to-face interviews were conducted with ten patients receiving care in a tertiary palliative care unit, a hospice or by palliative home care services in their homes. Several themes emerged. Emotional connection to others was an integral component of sexuality, taking precedence over physical expressions. Sexuality continues to be important at the end of life. Lack of privacy, shared rooms, staff intrusion and single beds were considered barriers to expressing sexuality in the hospital and hospice settings. Only one subject had previously been asked about sexuality as part of their clinical care, yet all felt that it should have been brought up, especially after the initial cancer treatments were completed. Home care nurses and physicians were seen as the appropriate caregivers to address this issue. Subjects unanimously mentioned that a holistic approach to palliative care would include opportunities to discuss the impact of their illness on their sexuality. PMID- 15540673 TI - A prospective study of the roles, responsibilities and stresses of chaplains working within a hospice. AB - Spiritual care is an integral part of palliative care and if asked, most members of a palliative care team would state they address spiritual issues. The majority of hospices have support from a chaplain. This study was to determine the roles of chaplains within hospices and to look at their levels of stress. A questionnaire containing both open and closed questions was sent to chaplains working within hospices in the UK. The questionnaire enquired about number of sessions, specific roles of chaplain, whether they were members of the multidisciplinary team and their sources of internal support. Stress was measured on a 10-point Likert scale and the GHQ12. One hundred and fifteen questionnaires were returned, with a 72% response rate. The majority (62%) defined their denomination as Church of England and Free Church (24%); 71% of respondents had parish commitments in addition to their hospice role. Roles were predominantly defined as spiritual care of patients and staff (95%) and bereavement support of relatives (76%) and 75% regularly attended the multidisciplinary meetings. Senior medical and nursing staff and other chaplains were perceived as providing most support. Median Likert score for stressfulness was 5, and 23% scored at or above the threshold on the GHQ12 for identifiable psychological morbidity. Clear role definition was associated with less perceived stress whereas the provision of bereavement support was associated with statistically significant increased perceived stress. The role of a chaplain within a hospice is varied and this study suggests that the provision of training and formal support is to be recommended. PMID- 15540674 TI - A competency model for the assessment and delivery of spiritual care. AB - The delivery of spiritual and religious care has received a high profile in national reports, guidelines and standards since the start of the millennium, yet there is, to date, no recognized definition of spirituality or spiritual care nor a validated assessment tool. This article suggests an alternative to the search for a definition and assessment tool, and seeks to set spiritual care in a practical context by offering a model for spiritual assessment and care based on the individual competence of all healthcare professionals to deliver spiritual and religious care. Through the evaluation of a pilot study to familiarize staff with the Spiritual and Religious Care Competencies for Specialist Palliative Care developed by Marie Curie Cancer Care, the authors conclude that competencies are a viable and crucial first step in 'earthing' spiritual care in practice, and evidencing this illusive area of care. PMID- 15540675 TI - A case for autopsy in palliative medicine? PMID- 15540676 TI - Cancer pain management in children. AB - Unrelieved pain may have a major impact on the care of children with cancer. The type and severity of pain experienced by children with cancer varies from acute, procedure-related pain to progressive chronic pain associated with the progression of the disease or sequelae of treatment. Drugs are the mainstay of treatment. Regular pain assessments combined with appropriate analgesic administration at regular dosing intervals, adjunctive drug therapy for control of adverse effects and associated symptoms, and nonpharmacological interventions are recommended. Although standard dosing of opioids adequately treats most cancer pain in children, more complex treatment is required by a significant group. Strategies to improve analgesia include the use of epidural or intrathecal infusions of a combination of opioids and other adjuvants, or other regional anaesthesia techniques. Procedure- and treatment-related pain is an even greater problem than cancer pain. Recommendations have been published with regard to the monitoring and personnel required when children are sedated which aim to set the standard of care and minimize both physical discomfort or pain and negative psychological responses, by providing analgesia; and to maximize the potential for amnesia; and to control behaviour. PMID- 15540677 TI - Transdermal fentanyl: caution in the use of high doses. PMID- 15540678 TI - Driving advice for palliative care patients taking strong opioid medication. PMID- 15540679 TI - Brain metastases and steroid reduction after radiotherapy. PMID- 15540680 TI - Audit of the use of a formatted postcard to communicate between a hospital palliative care team and general practitioners. PMID- 15540681 TI - Dental work as a cause of acute inflammation of a lymphoedematous limb. PMID- 15540682 TI - Raising the bar for the ethical integrity of clinical trials. PMID- 15540683 TI - Assessing the bioethical integrity of a clinical trial in surgery. PMID- 15540684 TI - Multivariate analysis of technical variables in pancreaticoduodenectomy: the effect of pylorus preservation and retromesenteric jejunal position on early outcome. AB - BACKGROUND: To evaluate the effect of technical modifications to pancreaticoduodenectomy (PD) on postoperative outcome, we established a register of all patients undergoing PD at Victoria General Hospital (Queen Elizabeth II Health Sciences Centre), a tertiary care, university-affiliated hospital. PATIENTS AND METHOD: Data from 78 consecutive patients who underwent PD from January 1998 through November 2000 were collected for univariate and multivariate analyses of clinical and technical factors on early outcome after PD, including duration of gastric stasis, development of complications and length of hospital stay. RESULTS: Two patients (2.6%) died; complications were recorded in 43 (55%). Upon univariate analysis, 3 factors (a diagnosis of chronic pancreatitis, pylorus preservation, and route of the jejunal limb) significantly affected duration of gastric stasis; but on multivariate analysis, only pylorus preservation and jejunal-limb route remained significant. Retromesenteric jejunal-limb placement was associated with longer periods of gastric stasis (mean 11.9 d, standard deviation [SD] 8.1 d) than the antemesenteric (retrocolic) route (mean 7.2, SD 3.6 d; p < 0.05); likewise pyloric preservation (mean gastric stasis 10.4 d, SD 5.9 d) compared with resection of the pylorus (mean 7.0 d, SD 3.2 d; p < 0.05). Pancreatic leaks occurred in 18% of retromesenteric and 8% of antemesenteric reconstructions (p = 0.3). Fewer patients with mucomucosal pancreaticojejunostomy suffered complications than those with invaginated anastomoses, but their hospital stays were similar in length. CONCLUSION: Route of the jejunal efferent limb and preservation of the pylorus are independent technical variables affecting early outcome after PD. PMID- 15540685 TI - Assessment of stability of the cervical spine in blunt trauma patients: review of the literature, with presentation and preliminary results of a modified traction test protocol. AB - OBJECTIVE: To review the reported efficacy of various imaging techniques in assessing stability of the neck in blunt trauma patients, and to present the protocol and preliminary results of a modified traction test protocol. DESIGN: This is a prospective cohort study. SETTING: A regional trauma unit in Southern Ontario. PATIENTS: People with blunt-trauma injuries who came to the author's consultant practice with "C-spine not cleared" status, from January 2001 through December 2003. INTERVENTIONS: A fluoroscopically controlled test of axial traction followed by flexion/extension stressing. OUTCOME MEASURES: Radiographic confirmation of the absence of pathological motion under load. RESULTS: In 51 cases studied to date, no instabilities have been found. Four cases of minor ligamentous hypermobility have been detected, with stability confirmed and no surgery required. There have been no failures to depict the neck completely, no missed instabilities and no complications of the procedure. CONCLUSIONS: Cervical stability can be reliably confirmed with this test without any requirement for advanced imaging technology. PMID- 15540686 TI - Results of cholecystectomy without intraoperative cholangiography. AB - BACKGROUND: To determine if cholecystectomy can be performed satisfactorily without the use of adjunctive intraoperative cholangiography (IOC), we planned a retrospective analysis at a Canadian university teaching hospital. METHODS: General operative morbidity and mortality (in particular, occurrences and complications of missed choledocholithiasis and reoperations for same, and occurrences of bile duct injuries and bile leaks) were noted and analyzed for a consecutive series of cholecystectomies from a single practice, carried out without IOC. MAIN RESULTS: In general, choledocholithiasis could be identified and treated before the operation; missed cases were infrequent and were treatable without reoperation. No major injuries to the bile duct were encountered. CONCLUSIONS: IOC appears to be optional with cholecystectomy; cholecystectomy can be performed without IOC safely in the defined setting, without related major complications from missed choledocholithiasis or excess occurrence of bile-duct injury. PMID- 15540687 TI - Magnetic resonance-guided percutaneous cryosurgery of breast carcinoma: technique and early clinical results. AB - PURPOSE: This phase I study was designed to demonstrate the feasibility, safety, efficacy and predictability of percutaneous cryosurgery, guided under magnetic resonance (MR) imaging, in the treatment of invasive breast carcinoma. PATIENTS AND METHODS: Under the guidance of near-real-time T1-weighted FSE images of a 0.5 T open-configuration MR system, percutaneous cryosurgery was performed in 25 patients with operable invasive breast carcinoma, 4 weeks prior to their scheduled mastectomy. Predictive assessments by interventional radiologists using 4 breast-imaging techniques (mammography, sonography, scintigraphy and MR) were correlated with postmastectomy results of histopathology and assessed for predictability. Local and systemic morbidity were also evaluated during the month of follow-up preceding mastectomy. RESULTS: Percutaneous cryosurgery resulted in no serious complications, either local or systemic. All tumoural tissues included in the cryogenic "iceball" were destroyed, with no viable histologic residues. Ablation was total in 13 of the 25 tumours treated. Combining periprocedural MR images with postprocedure scintimammographic findings enabled a 96% rate for predicting the cryosurgical results. CONCLUSIONS: MR-guided cryosurgery of breast carcinoma is feasible, safe and efficient, with predictable results. Major drawbacks are that the cryolesion (a palpable iceball) persists for a month or more after the procedure, undermining the reliability of the physical examination; and that breast imaging (mammography, ultrasound and MR) presents the same difficulty of interpretation as the physical exam even 1 month after the procedure. More studies are required to refine this treatment method. PMID- 15540689 TI - Is intensive care necessary after elective abdominal aortic aneurysm repair? AB - BACKGROUND: To review morbidity and mortality of patients undergoing elective, open repair of infra-renal abdominal aortic aneurysms and were admitted postoperatively to a surgical stepdown unit rather than routinely to the intensive care unit (ICU), we carried out a retrospective review. METHODS: All patients undergoing this type of repair in our centre, a division of vascular surgery in a tertiary-care teaching hospital in Ontario, over a 27-month period were reviewed. A consecutive 230 patients who underwent aneurysm repair from September 1999 through November 2001 were routinely admitted to a surgical stepdown unit postoperatively, with only a minority of patients requiring admission to ICU. We reviewed the rate of initial ICU admission and that of subsequent ICU admission after stepdown-unit admission. We also assessed morbidity, mortality and length of hospital stay for patients admitted to ICU as well as those admitted to the stepdown unit. RESULTS: ICU admission was avoided in 204 (89%) of these patients. The remaining 26 patients (11%) required ICU admission at some point during their hospital stay. Only 3 patients (1%) originally admitted to the stepdown unit subsequently required postoperative admission to ICU. CONCLUSIONS: Our experience demonstrates that proper preoperative assessment and selection allows the majority of elective infra-renal aneurysm repairs to be safely cared for postoperatively in a stepdown unit, and that subsequent ICU admissions are rare. PMID- 15540688 TI - Chronic lower extremity ischemia: a human model of ischemic tolerance. AB - BACKGROUND: Ischemic preconditioning (IPC) has been found in animals to have a protective effect against future ischemic injury to muscle tissue. Such injury is unavoidable during some surgical procedures. To determine whether chronic ischemia in the lower extremities would imitate IPC and reduce ischemic injury during vascular surgery, we designed a controlled clinical study. PATIENTS AND METHODS: Two groups of patients at a university-affiliated medical centre with chronic lower-extremity ischemia served as models of IPC: 6 patients awaiting femoral distal bypass (FDB) and 4 scheduled for aortobifemoral (ABF) bypass grafting for aortoiliac occlusive disease. Seven patients undergoing elective open repair of an infrarenal abdominal aortic aneurysm (AAA) were chosen as non IPC controls. Three hematologic indicators of skeletal-muscle injury, lactate dehydrogenase (LDH), creatine kinase (CK) and myoglobin, were measured before placement of the proximal clamp, during surgical ischemia, immediately upon reperfusion, 15 minutes after and 1 hour after reperfusion, and during the first, second and third postoperative days. RESULTS: Baseline markers of skeletal-muscle injury were similar in all groups. In postreperfusion samples, concentrations of muscle-injury markers were significantly lower in the 2 PC groups than in the control group. For example, at day 2, LDH levels were increased by about 30% over baseline measures in the elective AAA (control) group, whereas levels in the FDB and ABF groups remained statistically unchanged from baseline. Myoglobin in controls had increased by 977%, but only by 160% in the FDB and 528% in the ABF groups. CK levels, in a similar trend, were 1432% higher in the control group and only 111% (FDB) and 1029% (ABF) in the study groups. Taken together, these data represent a significant level of protection. CONCLUSIONS: Patients with chronic lower-extremity ischemia suffered less severe ischemic injury after a period of acute ischemia than those with acute ischemia alone. Ischemic preconditioning is one proposed mechanism to help explain this protective effect. PMID- 15540690 TI - Gastroduodenal tuberculosis management guidelines, based on a large experience and a review of the literature. AB - BACKGROUND: To review our experience of gastroduodenal tuberculosis before formulating management guidelines, we did a retrospective analysis at a large tertiary-care teaching institution in North India. METHOD: We reviewed 23 consecutive cases of biopsy-proven gastroduodenal tuberculosis over a period of 15 years. RESULTS: The major presenting features were gastric outlet obstruction (61%) and upper gastrointestinal (uGI) bleeding (26%). In 3 patients (13%), clinical, radiological and intraoperative features suggested malignancy/pseudotumour: periampullary mass in 2 and gastric mass in 1 patient. Five patients (23%) also had extragastrointestinal tuberculosis. Despite uGI endoscopy and biopsies, the preoperative diagnosis was correct for only 2 people. All patients except 1 required surgery for either diagnosis or therapy. Two patients with massive uGI hemorrhage requiring emergency surgery died in the postoperative period. The other patients responded well to antitubercular treatment after surgery. CONCLUSIONS: Gastroduodenal tuberculosis has 3 forms of presentation: obstruction, uGI bleeding, and gastric or periampullary mass suggestive of malignancy. Endoscopic biopsy has a poor yield. Surgery is usually required for diagnosis or therapy, after which patients respond well to antituberculous treatment. In areas endemic for tuberculosis, a good biopsy from the site of gastroduodenal bleeding or mass lesion and the surrounding lymph nodes should always be obtained. PMID- 15540691 TI - Surgical images: soft tissue. Mullerian cyst of the upper abdomen: a lesion mimicking pancreatic cystadenoma. PMID- 15540692 TI - Surgical Education and Self-Assessment Program (SESAP). A node in the anterior neck. PMID- 15540693 TI - Radiology for the surgeon. Musculoskeletal case 33. Post-traumatic osteolysis of the clavicle. PMID- 15540694 TI - Postsurgical tetanus. AB - The incidence of tetanus declined dramatically in the 20th century owing to routine vaccination and prompt attention to wound care. Postsurgical tetanus is uncommon, with both exogenous and endogenous sources being responsible for disease. The majority of cases of postoperative tetanus have been observed after intra-abdominal surgery. Those at high risk for developing tetanus include immigrants, the elderly, injection drug users, patients with diabetes and people of Hispanic ethnicity. Although most patients with tetanus can recover if managed appropriately, prevention through active and passive immunization is the main goal. This paper reviews postsurgical tetanus and provides an approach to its prevention and treatment. PMID- 15540695 TI - Treatment of lung hydatidosis by VATS: a preliminary report. PMID- 15540696 TI - Ischemic pancolitis: recognizing a rare form of acute ischemic colitis. PMID- 15540697 TI - Chylous ascites secondary to small-bowel angiosarcoma. PMID- 15540698 TI - Patellar tendon laxity in systemic lupus erythematosus. PMID- 15540699 TI - Combined trauma laparotomy and endovascular repair of thoracic aortic injury. PMID- 15540700 TI - Tibial plateau fractures. PMID- 15540701 TI - Mandatory reporting of gunshot wounds to police... Not as simple as it seems. PMID- 15540702 TI - Can informed consent be tailored for the critical care unit? PMID- 15540703 TI - Ontario court upholds regulatory authority of Ontario College of Pharmacists. PMID- 15540704 TI - Governments and health: visioning or repackaging? PMID- 15540705 TI - Ethical approval for national studies in Ireland: an illustration of current challenges. AB - BACKGROUND: Ethical approval of research projects is, appropriately, an essential prerequisite in health settings. AIMS: This paper outlines difficulties encountered with procedures for gaining ethical approval for two multicentre surveys in Ireland. METHODS: The experiences of two national surveys were documented. RESULTS: Delays in processing ethics applications led to substantial delays in both surveys. Research ethics committees (RECs) assessed applications in an idiosyncratic manner. CONCLUSION: In Ireland, there is currently no accepted mechanism for single location ethical approval for multicentre studies. Instead, they require separate approval from all participating centres. The challenges of this system of application to multiple committees are outlined in this paper, and possible solutions presented. PMID- 15540706 TI - Fifty years of carotid surgery--hail and farewell? AB - BACKGROUND: The operation of carotid endarterectomy (CEA) has had an eventful 50 years. AIM: This report reviews the literature comparing the outcome of CEA with that of medical therapy, angioplasty and stenting. METHODS: A review of the published randomised trials. RESULTS AND CONCLUSION: After 50 years, CEA is still the gold standard in the management of severe symptomatic carotid artery disease. It is doubtful, however, that in another 50 years it will continue to retain that pre-eminence. PMID- 15540707 TI - Patients' experiences of dialysis services: are national health strategy targets being met? AB - BACKGROUND: The National Health Strategy envisages a health system incorporating patient views; and providing accessible, consultant-led dialysis services with patient choice of dialysis modality, in all regions. AIMS: To describe patients' experiences of renal services against National Health Strategy objectives. METHODS: Telephone interviews with 192 dialysis patients from three hospitals in the Eastern region. RESULTS: One-quarter of participants (16% of haemodialysis [HD] and 46% of peritoneal dialysis patients) lived outside the Eastern region, and travelled there because dialysis was not available locally. Two-thirds (65%) had a choice of dialysis modality. High satisfaction with interpersonal care was observed (83-98% satisfaction). Dissatisfaction with physical environment included parking (39-56%), waiting areas (62-69%), HD unit space (74%). Regarding support services, dietary services were satisfactory (92-95%), with lower satisfaction ratings for social and financial support services (62%). CONCLUSIONS: Structural and management issues must be addressed to advance a quality agenda for renal care in Ireland. PMID- 15540709 TI - Risk factors for pulmonary embolism in an Irish patient cohort. AB - INTRODUCTION: The prevention of pulmonary embolism (PE) is an important component of medical care. AIM: To examine the risk factors for venous thromboembolism in an Irish patient cohort with acute PE, and identify cases that may have been preventable. METHODS: Retrospective review of 60 consecutive cases of computed tomography (CT)-confirmed acute PE. RESULTS: The primary thromboembolic risk factors were elective surgery (27%), medical illness (20%), primary immobility (13%) and isolated distal lower limb fracture (7%). A significant proportion (43%) had been hospitalised within the six weeks prior to PE onset. Some patients had undergone 'low risk' procedures, without prophylaxis, but had other significant thromboembolic risk factors that indicated a requirement for prophylaxis. CONCLUSIONS: Hospital- and ward-based thromboprophylaxis guidelines, based on certain categories of patient or procedure, need to be routinely supplemented by an individual risk factor assessment for each patient, to determine those at particularly high risk for venous thromboembolism. PMID- 15540708 TI - Impaired renal allograft, but not patient survival, in patients with antibodies to hepatitis C virus. AB - BACKGROUND: The impact of hepatitis C virus (HCV) infection in renal transplant patients is controversial and there are no data on the outcome of renal transplantation in this sub-group of Irish patients. AIM: To examine the outcome of renal transplantation in patients with hepatitis C. METHODS: We examined the outcome of first grafts from renal transplant patients with hepatitis C antibody positive and compared them to a control group. During this period, 24 HCV positive patients received 33 grafts. All were treated with standard immunosuppression. RESULTS: Graft survival rate was less in the HCV positive cases (p=0.0087). Graft survival at 1 year was 75% in the HCV positive group versus 85% in the HCV negative group, 40% versus 62% at 5 years and 14% compared with 40% at 10 years. Patient survival was similar in both groups (p=0.78). Patient survival at 1 year was 96% versus 94%, 87% versus 80% at 5 years and 70% in both groups at 10 years. CONCLUSION: In the Irish renal transplant population, the presence of hepatitis C antibodies, before or after transplantation is associated with worse long-term graft, but not patient survival. PMID- 15540710 TI - The cost of managing diabetic foot ulceration in an Irish hospital. AB - BACKGROUND: Little is known about the economic impact of diabetic foot ulceration in the Irish healthcare setting. AIM: Audit of diabetic foot ulcer admissions in St James's Hospital between April 2001 and March 2002. METHODS: Hospital charts were reviewed and costs were calculated on the length of patients' hospital stay and the cost of individual investigations performed. RESULTS: Thirty patients were admitted with diabetic foot ulceration as the primary complaint. Amputation was performed in eight patients, two patients with a non-healing ulcer died. The average duration of each hospital admission was 20.3+/-30.7 days. Net in-hospital expenditure was 704,689, an average of 23,489.63 per hospital admission. CONCLUSIONS: The management of diabetic foot ulceration has a significant economic impact on the Irish healthcare budget. Treatment should therefore be focused on primary prevention through specialised foot clinics and a multidisciplinary team approach to reduce this economic burden. PMID- 15540711 TI - Emergency department post-coital contraception in Northern Ireland. AB - BACKGROUND: The granting of a licence to Levonelle as an emergency hormonal contraceptive in the Republic of Ireland may require accident and emergency (A&E) departments to formally provide such a service. This article outlines the experiences of a Northern Ireland A&E unit. AIMS: To examine the pattern of attendance of patients requesting emergency contraception at an A&E department and to assess if adequate standards of care are achieved. METHOD: Retrospective case note review of 100 patients attending the A&E department requesting emergency contraception. RESULTS: Sixty-one per cent of requests for emergency contraception were outside normal pharmacy opening hours. Seventy-seven per cent of these patients were less than 26 years old. Most (63%) attended within 24 hours of unprotected sexual intercourse. Forty-three per cent of the patients studied had used no contraception prior to this request. Recording of menstrual details and sexual behaviour as part of the consultation was variable. CONCLUSIONS: A&E departments receive requests for emergency hormonal contraception particularly from younger women (<25 years). A&E staff must have appropriate training and support to manage these consultations effectively. PMID- 15540713 TI - Documentation of do-not-resuscitate orders in an Irish hospital. AB - BACKGROUND: Some studies have suggested that do-not-resuscitate (DNR) decisions are often documented poorly in European countries. AIM: To examine the use and documentation of DNR orders in a large Irish teaching hospital. METHODS: Resuscitation status of all inpatients on a single day was determined using interviews with nursing staff and examination of the nursing and medical case notes. RESULTS: Seventeen (3.5%) of 485 patients were identified as not for resuscitation. There was written confirmation of the DNR order in the nursing notes for 14 (82%) and in the medical notes for 15 (88%) patients; in two cases, it was reported that doctors were reluctant to write down the agreed decision. Documentation of DNR orders was by consultant (7), registrar (7) and intern (1). Discussion with patient (2), family (10) or both (1) was recorded in 14 cases. CONCLUSION: The majority of DNR orders were clearly documented by senior doctors and had been discussed with the patient or with the relatives. A number of problems were identified that might be avoided by development of guidelines regarding use and documentation of DNR orders. PMID- 15540712 TI - Detection of mycobacterial DNA from sputum of patients with cystic fibrosis. AB - BACKGROUND: Patients with cystic fibrosis (CF) are at high risk from atypical mycobacterial infections. There have been few attempts to delineate the intensity of mycobacterial infection in CF patients in Ireland. AIMS: To examine the incidence of mycobacterial DNA in an archived collection of genomic DNA extracted from the sputa of CF patients within the Northern Ireland population. METHODS: One hundred and eighty-two CF patients (66 adults and 116 children) were examined for the presence of mycobacterial DNA in their sputum by a genus specific PCR assay based on 16S rRNA, followed by direct automated sequencing of the PCR amplicons. RESULTS: One of 116 (0.9%) children and 2 of 66 adults were positive. Sequence identity revealed Mycobacterium xenopi in the paediatric patient and M. xenopi and M. chelonei in the two adult patients. False-positive results occurred in 11 patients (four adults), mainly due to Corynebacterium spp. CONCLUSIONS: There was a low prevalence of Mycobacterium spp in the CF patient population. All PCR positive results should be confirmed by direct automated sequencing and an alternative specific assay employed. Enhanced molecular screening will contribute in understanding their role as opportunistic pathogens in patients with worsening lung function. PMID- 15540714 TI - "Is this a dagger I see before me?"--an audit of stabbings and gunshot wounds in Limerick. AB - BACKGROUND: According to a recent study in Cardiff, the incidence of stab wounds is 14 per 100,000 population per annum. No such figures are available for Ireland. AIM: To evaluate the incidence, type of injury, medical consequences and outcome of patients with stab or gunshot wounds presenting to the Mid-Western Regional Hospital, Limerick, over a 12 month period. METHOD: A retrospective case study of all stab and gunshot wounds presenting over a 12 month period. RESULTS: Out of 62,000 new presentations to the Accident and Emergency (A&E) department, 101 (0.16%) were stabbings, giving an incidence of 33 per 100,000 population. Twenty-six patients required surgical intervention. There were three deaths. There were 10 gunshot wounds, of which 40% required surgical intervention, with no mortalities. CONCLUSION: The incidence of stab wounds presenting to our institution is high. Although constituting a small percentage of presentations to the A&E department they result in considerable morbidity and surgical activity. PMID- 15540715 TI - Neonatal transportation: the effects of a national neonatal transportation programme. AB - BACKGROUND: The transport of critically ill newborns by specialised transport teams has been shown to be associated with a significant improvement in their clinical condition on arrival at the receiving hospital. AIM: To determine if the National Neonatal Transport Programme introduced in 2001 improved clinical condition of newborns at the end of transfer. METHODS: A retrospective study of all 176 patients transported by the National Neonatal Transport Programme between March 2001 and March 2002. RESULTS: Before transfer, 17% of patients were hypothermic, 2% hypoglycaemic and 11% acidotic as were 7%, 3% and 5% respectively at the end of transfer. A review of 172 neonatal transports between 1987 and 1989 revealed that 21% of patients were hypothermic, 13% hypoglycaemic and 20% acidotic at the end of transfer. CONCLUSIONS: The National Neonatal Transport Programme has resulted in improved clinical condition of newborns at the end of transfer when compared to their condition before transfer and compared to outcomes prior to the introduction of the programme. PMID- 15540716 TI - Isolated Crohn's disease of the appendix. PMID- 15540717 TI - When is a cancer a malignancy? PMID- 15540718 TI - Racing for a cure. PMID- 15540719 TI - Stone treatments: current trends and future possibilities. PMID- 15540720 TI - Somatostatin analogues and estrogens in the treatment of androgen ablation refractory prostate adenocarcinoma. AB - PURPOSE: Prostate cancer progression to androgen ablation refractory stage D3 corresponds to cancer cell escape from androgen withdrawal induced apoptosis. Of note, salvage chemotherapy can extend the median survival of approximately 10 months in patients with stage D3. Therefore, novel therapeutic strategies that target the molecular basis of androgen resistance are required. MATERIALS AND METHODS: The MEDLINE and Current Content databases were used to find studies of the use of estrogens and somatostatin analogues for D3 prostate adenocarcinoma. We also analyzed the rationale and clinical results of our combination therapy using lanreotide and ethinylestradiol. RESULTS: Negative experiences have been reported with somatostatin analogues as monotherapy. On the other hand, the median progression-free survival reported in our experience using lanreotide acetate plus ethinylestradiol clearly surpassed the 10-month survival historically described in stage D3 cases. CONCLUSIONS: The use of somatostatin analogues in combination therapy for D3 prostate cancer sustains the novel concept in cancer treatment in which therapies may target not only cancer cells, but also the microenvironment in combination, which can confer protection from apoptosis. PMID- 15540721 TI - Molecular and cellular pathogenesis of benign prostatic hyperplasia. AB - PURPOSE: Symptomatic benign prostatic hyperplasia (BPH) is one of the most common ailments seen by the urologist. Significant advances have occurred in medical and surgical therapy, and in the understanding of the biology of this disease. However, the basic science literature is often conflicting and confusing, without a unified voice. We report the current state of knowledge of the molecular and cellular basis of BPH. MATERIALS AND METHODS: We compiled and interpreted basic science studies relevant to BPH pathogenesis. RESULTS: Cellular alterations that include changes in proliferation, differentiation, apoptosis and senescence in the epithelium and stroma are implicated in BPH pathogenesis. Molecular analyses have yielded numerous candidate genes important in disease progression. Differential expression of cytokines and growth factors in BPH tissue suggests roles for inflammation and hypoxia. Through the use of cell culture models the complex regulatory mechanisms of growth control in BPH are becoming defined. CONCLUSIONS: The scientific endeavor has resulted in great strides in our understanding of BPH on a molecular and cellular level. It is hopeful that basic science and translational research will improve treatment and prevention strategies for this common disease of elderly men. PMID- 15540722 TI - Is there a scientific basis for the therapeutic effects of serenoa repens in benign prostatic hyperplasia? Mechanisms of action. AB - PURPOSE: The huge resurgence of interest in herbal remedies has spawned a global industry that now competes with conventional drugs as adjuncts and/or alternatives for various conditions. The medical treatment of benign prostatic hyperplasia (BPH) is no exception. Along with alpha-blockers and 5alpha-reductase inhibitors the extract of the American dwarf palm, Serenoa repens, is unquestionably the most widely used. Together with Pygeum africanum, an extract from the bark of the African plum tree, it is licensed in Germany, France and other European countries for symptomatic BPH. This review was done to analyze the large number of in vivo and in vitro laboratory studies that have been performed with extracts of Serenoa repens to elucidate mechanism(s) of action. MATERIALS AND METHODS: A literature search (MEDLINE) revealed more than 30 publications relating to laboratory studies with extracts of Serenoa repens, addressing the question of a mechanism of action. It would appear that the n-hexane lipidosterolic extract of Serenoa repens, namely Permixon (Pierre Fabre Medicament, Boulogne, France), is a product that has uniquely been subjected to more scientific investigation than any other such preparation. RESULTS: Placebo controlled and comparative clinical studies of Permixon indicate its efficacy for BPH/lower urinary tract symptoms. Numerous mechanisms of action have been proposed, including an antiandrogenic action, an anti-inflammatory effect and an antiproliferative influence through the inhibition of growth factors. CONCLUSIONS: Set against the background of our current knowledge of the pathophysiology of the aging prostate, the results of these studies suggest a wide spectrum of activity. However, precise mechanism(s) of action remain obscure. Balance and caution are needed when extrapolating the results of in vitro laboratory studies to the complex human situation. PMID- 15540723 TI - Urological injuries in the Civil War. AB - PURPOSE: This study compiles all cases of urological injuries reported in the Civil War (1861 to 1865). MATERIALS AND METHODS: Using original sources largely assembled in army surgeon reports urological injuries documented and treated during the Civil War were recorded as to frequency, type, site and outcome. RESULTS: A total of 1,497 cases of injury involving the genitourinary organs were documented, representing 0.61% of all battle wounds, 22% of gunshot wounds of the abdomen and pelvis, and 47% of wounds restricted to the pelvis. Of these men 342 died (22% of all urological injuries and 37% of fatal pelvic wounds). Half of the kidney, bladder and prostate injuries were fatal, whereas men with injuries of the urethra, testes and penis generally recovered. Urethral wounds were often complicated by traumatic fistula and stricture. CONCLUSIONS: Wounds involving genitourinary organs and their consequences had a significant impact during the Civil War. As the war progressed, despite the limited means at their disposal surgeons learned how to better treat devastating urological injuries, resulting in improved survival and fewer severe complications. PMID- 15540724 TI - Oncological control following laparoscopic nephroureterectomy: 7-year outcome. AB - PURPOSE: Proof of the oncological safety of laparoscopic nephroureterectomy (LNU) relies on truly long-term outcome being at least equivalent to that of open surgery. We compared the long-term oncological outcome of laparoscopic versus open nephroureterectomy (ONU) in patients with upper tract transitional cell carcinoma (TCC). MATERIALS AND METHODS: Between April 1992 and January 1999, 26 LNUs and 42 ONUs were performed at our hospital for suspected upper tract TCC. Hospital medical records were retrospectively reviewed to assess preoperative staging, pathology and followup. RESULTS: There were 4 patients excluded from study (1 who underwent LNU and 3 ONU) since the histological diagnosis was other than TCC. Median followup for the laparoscopic and open groups was 101 and 96 months, respectively. There was local recurrence in 2 patients (8%) after LNU and in 6 patients (15.4%, p = 0.3) after ONU. TCC recurred in the contralateral kidney or ureter in 2 LNU cases (8%) and 1 ONU case (2.6%, p = 0.3). There was bladder recurrence in 7 patients (28%) following LNU compared with 15 patients (42%, p = 0.2) after open nephroureterectomy. The 1 and 5-year metastasis-free survival rates were 80% and 72% for LNU compared with 87.2% and 82.1% for ONU (p = 0.33 and 0.26). Upper tract tumor grade and stage influenced the incidence of metastatic and contralateral disease, but not the incidence of local or bladder recurrence. CONCLUSIONS: In the surgical management of upper tract TCC, the laparoscopic approach does not affect long-term oncological control. Tumor stage and grade are important prognostic factors in the development of metastases and cancer specific mortality. PMID- 15540725 TI - Laparoscopic extended pelvic lymphadenectomy for bladder cancer: technique and initial outcomes. AB - PURPOSE: We describe the technique and our evolving outcomes of laparoscopic extended pelvic lymphadenectomy for bladder cancer. MATERIALS AND METHODS: Since 1999 laparoscopic radical cystectomy with pelvic lymphadenectomy and intracorporeal urinary diversion has been performed in 22 patients. The initial 11 patients underwent limited dissection (group 1) and the subsequent 11 consecutive patients underwent extended lymphadenectomy (group 2). Our split-and roll technique of laparoscopic extended pelvic lymphadenectomy has evolved to achieve lymphatic tissue clearance by bilaterally skeletonizing the genitofemoral nerve, external iliac artery, external iliac vein, obturator nerve, hypogastric artery, common iliac artery and pubic bone. RESULTS: Extended lymphadenectomy added 1.5 hours of operative time. The median number of nodes retrieved was 3 and 21 in groups 1 and 2, respectively (p = 0.001). Three patients per group were found to have positive nodal disease. In 1 patient undergoing extended dissection injury to a deep pelvic vein was managed by intracorporeal suturing and resulted in 200 ml blood loss. Two other patients had deep venous thrombosis. At a mean followup of 11 months (range 2 to 43) there were no port site recurrences. CONCLUSIONS: Laparoscopic extended pelvic lymphadenectomy for bladder cancer can be performed with anatomical boundaries and nodal yields commensurate with those of current recommendations for open surgery. PMID- 15540726 TI - Intravesical explosion during transurethral resection of bladder tumors. PMID- 15540727 TI - Assessment of perioperative psychological distress in patients undergoing radical cystectomy for bladder cancer. AB - PURPOSE: Despite a recent growth in our understanding of the impact of psychosocial factors on the outcome of patients with cancer there is still relatively little known about the effect of these issues on patients with genitourinary malignancies. We determined the prevalence of psychological distress in patients with bladder cancer prior to and following radical cystectomy. MATERIALS AND METHODS: A total of 74 consecutive patients with clinically organ confined bladder cancer were prospectively surveyed preoperatively using the Basic Symptom Inventory-18, a validated instrument that measures the psychological domains of general distress, anxiety, depression and somatization. Of the initial 74 patients 62 were available for postoperative assessment 1 month following cystectomy. Preoperative and postoperative distress scores were evaluated with respect to age, sex, marital status, type of surgical reconstruction and tumor stage. RESULTS: The preoperative prevalence of psychological distress in patients diagnosed with bladder cancer was 45% and it remained somewhat increased at 34% approximately 4 weeks after cystectomy. Demographic factors such as gender, age, and marital status were not significantly associated with the overall prevalence of distress. In the entire study group there was a statistically significant decrease in general distress (p = 0.028), depression (p = 0.034) and anxiety (p = 0.0004) from the preoperative to the postoperative assessments. Pathological stage was significantly associated with post-cystectomy anxiety (p = 0.040) and general distress (p = 0.042). CONCLUSIONS: Our findings indicate that a large proportion of patients with bladder cancer undergoing radical cystectomy experience psychological distress during the perioperative period. The identification of psychological distress in this population has the potential to influence health related quality of life as well as recovery in all individuals with bladder cancer. PMID- 15540728 TI - Lymph node involvement in patients with bladder cancer treated with radical cystectomy: a patho-anatomical study--a single center experience. AB - PURPOSE: To our knowledge the extent of lymphadenectomy with cystectomy, the number of lymph nodes to be retrieved and the anatomical groups to be dissected are still undetermined. This study was done to clarify these issues. MATERIALS AND METHODS: A total of 200 patients underwent radical cystectomy and extended lymphadenectomy up to the level of origin of the inferior mesenteric artery. Removed tissues were labeled according to anatomical location and sent separately for pathological evaluation. In each group the number and status of lymph nodes were determined. The number of positive nodes was correlated with the number of retrieved nodes. Cases with a single positive node were identified and the anatomical location was defined. RESULTS: The mean number of retrieved nodes per patient +/- SE was 50.6 +/- 14.4 and 48 (24%) patients had nodal disease. The mean number of positive nodes per involved case was 8.08 +/- 13.2. There was a weak correlation between the number of positive nodes and the number of harvested nodes. Bilateral disease was noted in 39.6% of cases. Single node involvement was observed in 22 cases, of which all except 1 were in the endopelvic region. CONCLUSIONS: There is a sentinel region, which is the endopelvic region (that is the internal iliac and obturator groups of lymph nodes). There are no skipped lesions. Negative nodes in the endopelvic region indicate that more proximal dissection is not necessary. Bilateral endopelvic dissection is mandatory. PMID- 15540729 TI - Controversies in prostate cancer screening. AB - PURPOSE: Prostate cancer is the most common cancer in men and the second most common cause of cancer death in men, and yet controversy continues to surround the practice of prostate cancer screening. Despite recent studies that have cast doubt over the true efficacy of breast cancer screening programs, the National Cancer Institute continues to support breast cancer screening and it has withheld endorsement of widespread prostate cancer screening. Criticisms of prostate cancer screening include the financial burden of screening, the morbidity of prostate biopsy, the low positive predictive value of screening, the over treatment of an indolent disease and the lack of evidence demonstrating a mortality benefit due to screening. MATERIALS AND METHODS: We formulated a comprehensive discussion addressing the criticisms of prostate cancer screening. RESULTS: In an effort to highlight the importance of prostate cancer screening we noted how concerns regarding cost, morbidity and low positive predictive value are common to widely accepted screening programs for other common malignancies. We also draw attention to the danger of abandoning prostate cancer screening, a practice that is called into question by watchful waiting series and Markov modeling of prostate cancer treatment. Finally, we observed how the implementation of prostate cancer screening in the United States has led to the phenomenon of stage migration and paralleled the decrease in the prostate cancer mortality rate. CONCLUSIONS: The prostate specific antigen era has brought great promise for improving the prognosis of prostate cancer. We must continue to seek support for widespread prostate cancer screening. PMID- 15540730 TI - Should an isolated humeral lesion detected during prostate cancer metastatic survey be biopsied? PMID- 15540731 TI - Thyroid uptake of 125iodine after prostate permanent brachytherapy. AB - PURPOSE: To decrease the seed migration rate after 125I prostate brachytherapy and improve the quality of implants to our knowledge we report for the first time the detection of thyroid uptake, possibly from a damaged seed, in a patient after 125I implantation. MATERIALS AND METHODS: Seed migration detection is routinely offered to our patients at the post-evaluation visit. A seed migration detector and comprehensive investigation procedure have been developed for the task. Chest radiographs are ordered to document the location of the detected seed. A fluoroscopic study may be performed if intracardiac radioactivity is detected. RESULTS: Since November 2001, 246 patients have been studied for seed migration. A total of 23,184 125I seeds were implanted, of which 75 were lost through the urethra and 25 migrated to the thorax. In the routine survey of a patient on February 13, 2003 radioactivity in the chest-neck region was sensed. Repeat radiographs and fluoroscopic examinations were negative for seed migration. Using a gamma camera butterfly-shaped uptake was noted in the thyroid region. The energy peak of the uptake matched the spectrum of the 125I source. The patient denied any intake of other iodine medication. The results suggested possible 125I leakage from at least 1 seed implanted in the prostatic region. CONCLUSIONS: Improved control of seed migration has been achieved. The detection of thyroid uptake is reported. Detection techniques and subsequent actions are described. PMID- 15540732 TI - Watchful waiting and health related quality of life for patients with localized prostate cancer: data from CaPSURE. AB - PURPOSE: Watchful waiting is an alternative to active treatment for men with low risk prostate cancer but it is unclear how health related quality of life (HRQoL) may change over time for men who select this option. We report on HRQoL in men with localized prostate cancer who selected watchful waiting. MATERIALS AND METHODS: HRQoL outcomes were reviewed for 310 men diagnosed with prostate cancer from 1990 to 2001 within Cancer of the Prostate Strategic Urological Research Endeavor who chose watchful waiting. The UCLA Prostate Cancer Index and RAND 36 Item Health Survey were completed at enrollment and approximately every 6 months. A random slopes model was developed to assess time trends in HRQoL for up to 5 years after diagnosis, adjusting for age at diagnosis and specific comorbidities. RESULTS: Significant decreases with time were observed in 7 domains of the RAND 36-Item Health Survey and 4 of the UCLA Prostate Cancer Index scales. CONCLUSIONS: Men with prostate cancer who chose watchful waiting in the current study had better or similar HRQoL outcomes compared to men without prostate cancer at the start of the study. Many of these scores were significantly affected by increasing age and decreased with time. The physical domain scores as well as sexual function scores decreased more than expected from the aging process alone. PMID- 15540734 TI - Anatomical extent of lymph node dissection: impact on men with clinically localized prostate cancer. AB - PURPOSE: This study evaluates the influence of the anatomical extent of pelvic lymph node dissection performed at radical prostatectomy on lymph node yield, staging accuracy and time to prostate specific antigen progression. MATERIALS AND METHODS: Between February 1992 and April 2003, 2 surgeons at 1 hospital performed 2,135 and 1,865 radical prostatectomies with pelvic lymph node dissection, respectively. One surgeon routinely performed an extended lymph node dissection while the second surgeon performed a limited pelvic lymphadenectomy. The number of lymph nodes extracted and the number of patients with positive lymph nodes detected were analyzed and compared. Kaplan-Meier analysis was used to compare the biochemical recurrence-free survival between the 2 groups of patients with occult nodal disease. RESULTS: Extended lymph node dissection removed more lymph nodes (mean 11.6 vs 8.9, p<0.0001) and detected more lymph node positive disease (3.2% vs 1.1%, p<0.0001) than the more anatomically limited technique. This finding held true for patients across all pathology groups. Among men with lymph node positive disease involving less than 15% of extracted nodes, the 5-year prostate specific antigen progression-free rate for extended lymph node dissection was 43% versus 10% for the more limited lymph node dissection (p = 0.01). CONCLUSIONS: Compared to limited lymph node dissection, extended pelvic lymphadenectomy appears to identify men with positive lymph nodes more frequently. A significant benefit in biochemical recurrence-free survival may exist for certain subgroups undergoing the extended dissection. However, because the results may be influenced by stage migration, longer followup is necessary to determine whether the apparent therapeutic effect persists. PMID- 15540733 TI - Impact of surgical delay on long-term cancer control for clinically localized prostate cancer. AB - PURPOSE: Radical retropubic prostatectomy (RRP) as definitive management for clinically localized prostate cancer is commonly performed within months of diagnosis. Despite patient anxiety there is little evidence to suggest that a delay of several months from diagnosis to RRP is associated with a worse cancer control rate. However, a recent study cast doubt on the safety of such a delay with respect to cancer control. Therefore, in a contemporary series we determined long-term cancer control in men who underwent RRP for clinically localized prostate cancer with some treated early and others treated after a longer delay. MATERIALS AND METHODS: We analyzed data on 926 men who underwent RRP between January 1989 and December 1994. Age, preoperative serum prostate specific antigen (PSA), biopsy Gleason score, clinical and pathological stage, and biochemical recurrence were compared between 162 men who underwent RRP 60 days or less from biopsy and 764 who underwent RRP after a greater delay. Disease-free (PSA less than 0.2 ng/ml) survival rates were compared using Kaplan-Meier analysis. Pathological staging was compared using logistic regression. RESULTS: The different groups were well matched for age, serum PSA, pathological stage and followup. However, significantly more men who underwent RRP between 121 and 150 days, and 151 days or greater had T1c disease (48% and 57% vs 35%, p<0.04 and <0.0001, respectively). In addition, significantly more men operated on at 151 days or greater had biopsy Gleason scores 2 to 6 (86% vs 65%, p<0.0001) and significantly fewer had Gleason score 7 disease (13% vs 30%, p<0.002). Men who underwent RRP after 60 or less days had 5 and 10-year biochemical disease-free survival rates comparable to those in men who underwent RRP after 61 to 90, 91 to 120 and 121 to 150 days after diagnosis (82% and 78%, 86% and 78%, 86% and 75%, and 86% and 82%, respectively). Those operated on at 151 days or greater had significantly greater 5 and 10-year biochemical disease-free survival rates (89% and 87%, p<0.04). However, when patients were stratified into different subgroups based on clinical stage, serum PSA and biopsy Gleason score a delay of 150 days or greater no longer impacted differently on long-term cancer control rates. CONCLUSIONS: Delays of up to several months from prostate cancer diagnosis to RRP do not appear to impact long-term biochemical cancer control rates. Therefore, patients can be reassured that there is no immediate urgency to perform RRP after prostate cancer diagnosis, especially in those with T1c disease and biopsy Gleason scores less than 7. PMID- 15540735 TI - Impact of margin size on the incidence of local residual tumor after laparoscopic radical prostatectomy. AB - PURPOSE: Since October 2001 we have performed intraoperative frozen section (IFS) on the posterolateral areas of the radical prostatectomy specimen with bilateral nerve sparing. In cases of positive surgical margin (SMs) the corresponding neurovascular bundle (NVB) was resected. We evaluated the impact of several biological and pathological parameters on the incidence of local residual tumor in the NVB. MATERIALS AND METHODS: A total of 487 laparoscopic radical prostatectomies with bilateral nerve sparing and IFS were performed between October 2001 and December 2003. When IFS was positive, the whole NVB was removed and analyzed after fixation in serial sections. The size of the positive SM was measured in its largest dimension. The association of NVB residual tumor with positive SM size, preoperative prostate specific antigen, pathological stage and Gleason score was analyzed using the t and chi-square tests, and logistic regression. RESULTS: Of the 487 patients 84 had a positive SM on IFS. In 25 of the 84 additionally resected NVBs we found residual tumor. On univariate and multivariate analysis the only predictor of residual tumor was positive SM size (p<0.001). In all cases of a positive SM of 0.1 cm or less we found no residual tumor cells in the NVB. CONCLUSIONS: In case of a positive SM in the posterolateral area of the prostatectomy specimen margin size is predictive of the incidence of residual tumor in the corresponding NVB. This finding could help management and the decision about local adjuvant treatment. PMID- 15540736 TI - CYP3A4 genetic variant and disease-free survival among white and black men after radical prostatectomy. AB - PURPOSE: Prostate cancer is an androgen sensitive disease. Cytochrome P450 3A4 (CYP3A4) oxidatively deactivates testosterone by converting it to biologically less active metabolites. Previous studies suggest that a germline genetic variant in the 5' regulatory region of the gene may interfere with deactivation and increase the risk of clinically advanced prostate cancer. We investigated the impact of this polymorphism on the risk of recurrence after prostatectomy. MATERIALS AND METHODS: We assembled clinical data and analyzed specimens from a large series of patients who underwent prostatectomy who were carefully staged at a single institution and had 5 to 10 years of prospective clinical followup. The series included 428 white men and 309 black men. RESULTS: Stage, Gleason score or preoperative prostate specific antigen strongly predicted progression-free survival (PFS) but were not associated with CYP3A4 genotypes. There was a strong association between race and genotype (p = 0.00002) in that 8% of white men and 83% of black men had 1 or more copies of the G allele. When both races were included genotype was associated with PFS (hazard ratio [HR] 1.27, CI 1.08-1.27, p = 0.005). In race specific analyses increasing copies of the G allele were associated with poorer PFS among white men (HR 1.98, CI 1.06-3.70, p = 0.03) but had little impact on PFS among black men (HR 1.004, CI 0.77-1.32, p = 0.97). CONCLUSIONS: The CYP3A4 genotype studied was not associated with pathological features of prostate cancer for men of either race. Unstratified analyses of men of both races and stratified analyses of white men demonstrated poorer PFS after prostatectomy for those with the G allele, but the G allele did not predict PFS among black men. PMID- 15540737 TI - Race is not a predictor of prostate cancer detection on repeat prostate biopsy. AB - PURPOSE: We evaluated men undergoing repeat prostate biopsies for persistently increased serum prostate specific antigen (PSA) levels to determine if race was a predictor of cancer detection. MATERIALS AND METHODS: Between July 1995 and June 2002, 401 men had undergone 2 or more transrectal ultrasound guided prostate biopsies at our institutions. Clinical information was gathered using our prostate biopsy database and retrospectively reviewed. Race, age, serum PSA, PSA velocity, total number of biopsies performed, total number of previous negative cores and the presence of high grade prostatic intraepithelial neoplasia (HGPIN) or atypical small acinar proliferation (ASAP) on prior biopsy were evaluated to determine if they were predictors of subsequent cancer detection. Multivariate analysis was performed using a time dependent covariate Cox proportional hazards model. RESULTS: Of the 401 men undergoing repeat prostate biopsy, 91 (22.7%) were diagnosed with prostate cancer. In total there were 180 (44.9%) black men and 221 (55.1%) white men. Cancer was diagnosed in 49 black men (27.2%) and 42 white men (19.0%, p = 0.06). On multivariate analysis serum PSA, HGPIN, ASAP and PSA velocity were predictors of prostate cancer detection (p = 0.006, <0.0001, 0.001 and 0.0004, respectively). Race was not found to be a predictor of prostate cancer detection on repeat prostate biopsy (p = 0.16). In the evaluation of clinical data for racial differences, black men had a significantly higher incidence of HGPIN on prior biopsy compared to white men (p = 0.02). Serum PSA, PSA velocity, presence of ASAP on prior biopsy, age, number of biopsies performed and number of previous negative cores were not statistically different between black and white men. CONCLUSIONS: Race is not a predictor of prostate cancer detection in men undergoing repeat prostate biopsies. With the exception of HGPIN, all other clinical parameters were similar between black and white men. Serum PSA, PSA velocity, HGPIN and ASAP were found to be significant predictors of subsequent prostate cancer detection. PMID- 15540738 TI - Is Hispanic race an important predictor of treatment failure following radical prostatectomy for prostate cancer? AB - PURPOSE: Hispanic-Americans are the fastest growing minority group in the United States. Many studies have compared prostate cancer treatment outcomes between black and white men, but few such studies have been done with Hispanic men. We compared clinical and pathological features as well as the treatment failure rate of radical prostatectomy in contemporaneously treated groups of Hispanic and white men with prostate cancer. MATERIALS AND METHODS: Between 1995 and 2002, 136 Hispanic men and 315 white men underwent radical prostatectomy. Treatment failure was defined as having a prostate specific antigen (PSA) of 0.2 or greater more than 8 weeks after surgery or receiving any adjuvant therapy. Known predictors of failure and race were evaluated for their ability to predict treatment failure. RESULTS: Median followup was 32 months for Hispanic and 36 months for white patients. Hispanic men were older, had a higher percentage of abnormal rectal examinations, Gleason 7 tumors and preoperative PSA levels greater than 10. Preoperative PSA, specimen Gleason score, pathological stage and surgical margin were all strongly associated with treatment failure (p<0.001). Despite differences in clinical characteristics, overall failure rates did not differ between Hispanic and white men (18.7% vs 17.8%). The odds ratio for treatment failure for Hispanic relative to white men after adjusting for the previously mentioned risk factors was 0.87 (95% CI [0.44, 1.68], p = 0.670). CONCLUSIONS: This study shows that Hispanic race does not influence the treatment failure rate of radical prostatectomy in contemporaneously treated patients with prostate cancer at 1 institution. To our knowledge this study represents the largest of its kind, but longer followup and other confirmatory studies are needed. PMID- 15540739 TI - Prostate specific antigen progression in men with lymph node metastases following radical prostatectomy: results of long-term followup. AB - PURPOSE: We examined the clinical outcome of patients with lymph node metastases found at prostatectomy with the goal to identify factors that predict freedom from prostate specific antigen (PSA) progression. MATERIALS AND METHODS: We retrospectively reviewed the records of 3,264 consecutive men with clinically localized prostate cancer who underwent extended pelvic lymphadenectomy and radical prostatectomy performed by a single surgeon between April 1982 and March 2003. Patients with pathologically confirmed lymph node metastases and no history of adjuvant treatment were identified. Clinical and histopathological factors were analyzed for an association with time to PSA progression using univariate and multivariable analyses. RESULTS: Of the 143 patients (4.4% of total) in the study with nodal involvement 24 (16.8%) were free of disease at last followup (median 6 years). Median time to failure was 2 years with PSA progression occurring as late as 11 years postoperatively in 2 patients. The 5 and 7-year PSA progression free rate in all lymph node positive patients was 26.5% and 10.9%, respectively. A 15% or greater incidence of positive nodes (p = 0.0008) and high prostatectomy Gleason score (ie score 8 to 10, p = 0.008) were independent predictors of PSA progression in multivariate Cox proportional hazards models. Seminal vesicle invasion (HR 1.45, p = 0.063) or positive surgical margins (HR 1.43, p = 0.063) were marginally significant in the multivariate model. The 5 year PSA progression-free rate was 52% in men with less than 15% positive lymph nodes, prostatectomy Gleason score 7 or less and negative seminal vesicle invasion. CONCLUSIONS: While the incidence of lymph node positive disease in patients undergoing radical prostatectomy is infrequent in the PSA era, patients with nodal involvement may experience disease progression as remote as 1 decade after surgery. Pathological factors such as the percent of positive lymph nodes, prostatectomy Gleason score and seminal vesicle invasion appear to predict an increased risk of PSA failure in this population. PMID- 15540740 TI - Bicalutamide 150 mg in addition to standard care in patients with localized or locally advanced prostate cancer: results from the second analysis of the early prostate cancer program at median followup of 5.4 years. AB - PURPOSE: We evaluated the efficacy and tolerability of 150 mg bicalutamide daily given in addition to standard care, in patients with localized or locally advanced prostate cancer. MATERIALS AND METHODS: The bicalutamide Early Prostate Cancer program consists of 3 randomized, double blind, placebo controlled trials prospectively designed for combined analysis. A total of 8,113 men with T1b-T4, M0, any N (N0 in 1 trial) prostate cancer were randomized to bicalutamide 150 mg/day (4,052) or placebo (4,061) in addition to standard care (radical prostatectomy, radiotherapy or watchful waiting). Primary end points were objective progression-free survival (PFS) and overall survival. RESULTS: At median 5.4 years of followup (21.6% progression events) bicalutamide significantly improved PFS in the overall population. This result was driven by positive results in trials 24 and 25, with the North American trial (trial 23) showing no difference. Patients with locally advanced disease gained most benefit from bicalutamide in terms of PFS, irrespective of underlying therapy. Overall survival was similar in the bicalutamide and placebo groups, across the program and in each trial. Among watchful waiting patients survival appeared to be improved with bicalutamide in those with locally advanced disease, whereas survival appeared to be reduced with bicalutamide in those with localized disease. The most common adverse events with bicalutamide were gynecomastia and breast pain. Other adverse events occurred with a similarly low incidence in the 2 treatment groups. CONCLUSIONS: This analysis confirms that bicalutamide provides benefit in patients with locally advanced disease. The current data suggest that early or adjuvant hormonal therapy for patients at low risk of disease progression, such as those with localized disease, is not appropriate. PMID- 15540741 TI - Bicalutamide (150 mg) versus placebo as immediate therapy alone or as adjuvant to therapy with curative intent for early nonmetastatic prostate cancer: 5.3-year median followup from the Scandinavian Prostate Cancer Group Study Number 6. AB - PURPOSE: We evaluated the benefits of adding 150 mg bicalutamide to standard care, that is radical prostatectomy, radiotherapy or watchful waiting (WW), in patients with localized or locally advanced prostate cancer. MATERIALS AND METHODS: A total of 1,218 men with T1-4, M0, any N prostate cancer were recruited from 62 Scandinavian centers and randomized 1:1 to 150 mg bicalutamide or placebo plus standard care. Primary end points were progression-free survival (PFS) and overall survival. RESULTS: At a median 5.3-year followup patients with locally advanced disease had improved survival with bicalutamide (HR 0.68, 95% CI 0.50 to 0.92), while those with localized disease had decreased survival with bicalutamide (HR 1.47, 95% CI 1.06 to 2.03). Bicalutamide significantly improved PFS, decreasing the risk of disease progression by 43% compared with placebo (HR 0.57, 95% CI 0.48 to 0.68, p<0.0001). The rate of events was 35.4% for bicalutamide and 46.2% for placebo. Patients with locally advanced disease gained the greatest PFS benefits with bicalutamide (HR 0.40, 95% CI 0.31 to 0.52). Since 81% of the trial population were untreated before entry and would otherwise have undergone WW, the findings essentially reflect the results of immediate hormone therapy vs WW. CONCLUSIONS: Bicalutamide (150 mg) provides significant benefit in patients with locally advanced disease. In previously untreated patients there may be a tumor burden below which endocrine therapy provides no benefit or may even decrease survival. PMID- 15540742 TI - Squamous cell carcinoma of the penis presenting as a sterile inguinal abscess. PMID- 15540743 TI - Treatment of acute inguinal hemorrhage from metastatic penile carcinoma using an endovascular stent graft. PMID- 15540744 TI - Transdermal posterior tibial nerve laser therapy is not effective in women with interstitial cystitis. AB - PURPOSE: Interstitial cystitis (IC) is a debilitating condition which causes irritative bladder symptoms, pain and a decrease in health status. The pathophysiology is poorly understood so therapeutic options are diverse. Percutaneous posterior tibial nerve stimulation is an effective treatment and pulsed transdermal laser stimulation is an established technique for pain management. We evaluated the efficacy of transdermal laser stimulation of the posterior tibial nerve for patients with IC. MATERIALS AND METHODS: Women meeting the National Institutes of Health National Institute for Diabetes and Digestive and Kidney Diseases criteria for IC were prospectively recruited and randomized to treatment (29) or placebo (27) cohorts in a double-blind trial. At home the patient performed laser therapy daily for 30 seconds over the SP6 acupuncture point for 12 weeks. Measures at baseline and at 84-day followup included the 7 day voiding diary, the Interstitial Cystitis Problem Index, Interstitial Cystitis Symptom Index and RAND 36-Item Health Survey questionnaires. RESULTS: There were no significant differences between the treatment and control cohorts on any of the measures. However, there was a significant decrease between baseline and 12 week followup in the amount voided, symptom problems and severity, and on all 8 SF-36 scales. There was no significant effect of fluid intake. CONCLUSIONS: This study demonstrated no difference between the active and sham device. However, it is interesting that treatment and control cohorts experienced similar improvements, suggesting that the control cohort improvements may have been due to participants' belief that they were receiving active treatment from the stimulator. These findings provide support for investigating placebo effects in randomized trials. PMID- 15540745 TI - Simple solution for urethral caruncle. AB - PURPOSE: Although urethral caruncle is not a major disease, the technique to remove it has been rather difficult and complicated. We describe a simple method of removal that is comparable to its importance. MATERIALS AND METHODS: In 10 patients we applied a ligation technique using 1-zero silk at the base of the urethral caruncle during a 4-year period. RESULTS: This technique was well tolerated by patients without analgesics or anesthesia. It also allowed obtaining tissue pathological evaluation if wanted. Seven to 14 days after ligation the caruncles broke off without any problem. The caruncles base healed satisfactorily. However, this technique requires at least 1 assistant. CONCLUSIONS: We describe a simple treatment for urethral caruncle. PMID- 15540746 TI - Intermittent testicular torsion in a 58-year-old man. PMID- 15540747 TI - Comparison of 2 generations of piezoelectric lithotriptors using matched pair analysis. AB - PURPOSE: The Piezolith 3000 (Richard Wolf, Knittlingen, Germany) is the newest piezoelectric lithotriptor. Using a matched pair analysis model we compared treatment outcomes of this machine with those of an older generation Piezolith 2300 lithotriptor (Richard Wolf). MATERIALS AND METHODS: Patients with solitary, radio-opaque urinary calculi undergoing primary lithotripsy using the Piezolith 3000 were identified. All patients had a 3-month followup. These patients were matched with those from our database treated between 1992 and 1999 with a Piezolith 2300. Patients were initially matched for gender, side and site of the stone. For stones other than those in the lower calix and lower ureter, matching was performed for size in terms of maximum and minimum diameter of the index stone. Additional anatomical factors of caliceal pelvic height and vertical distance of the lower ureteral stone from pubic symphysis were also measured. Thereafter stones with size +/- 1 mm of the index stone were selected and the stone with the best matched anatomical factors was chosen. The initial stone fragmentation rate and stone-free rates at 3 months in the matched pairs were then compared. RESULTS: A total of 25 matched pairs were found between October 2002 and December 2002. There was no statistical difference between the initial fragmentation rate for the Piezolith 3000 (68%) and the Piezolith 2300 (84%, McNemar's test p = 0.388). The stone-free rate at 3 months for the Piezolith 3000 and the Piezolith 2300 were 36% and 48%, respectively, again with no statistical difference (McNemar's test p = 0.581). CONCLUSIONS: Despite significant design changes and technical modifications, the new piezoelectric lithotriptor does not appear to provide a better treatment outcome than the older generation machine. PMID- 15540748 TI - Innovations in shock wave lithotripsy technology: updates in experimental studies. AB - PURPOSE: We developed innovations in shock wave lithotripsy (SWL) technology. MATERIALS AND METHODS: Two technical upgrades were implemented in an original unmodified HM-3 lithotriptor (Dornier Medical Systems, Inc., Kennesaw, Georgia). First, a single unit ellipsoidal reflector insert was used to modify the profile of lithotriptor shock wave (LSW) to decrease the propensity of tissue injury in SWL. Second, a piezoelectric annular array (PEAA) generator (f = 230 kHz and F = 150 mm) was used to produce an auxiliary shock wave of approximately 13 MPa in peak pressure (at 4 kV output voltage) to intensify the collapse of LSW induced bubbles near the target stone for improved comminution efficiency. RESULTS: Consistent rupture of a vessel phantom made of single cellulose hollow fiber (i.d. = 0.2 mm) was produced after 30 shocks by the original HM-3 reflector at 20 kV. In comparison no vessel rupture could be produced after 200 shocks using the upgraded reflector at 22 kV or the PEAA generator at 4 kV. Using cylindrical BegoStone phantoms (Bego USA, Smithfield, Rhode Island) stone comminution efficiencies (mean +/- sd) after 1,500 shocks produced by the original and upgraded HM-3 reflectors, and the combined PEAA/upgraded HM-3 system, were 81.3% +/- 3.5%, 90.1% +/- 4.3% and 95.2% +/- 3.3%, respectively (p<0.05). CONCLUSIONS: Optimization of the pulse profile and sequence of LSW can significantly improve stone comminution while simultaneously decreasing the propensity of tissue injury during in vitro SWL. This novel concept and associated technologies may be used to upgrade other existing lithotriptors and to design new shock wave lithotriptors for improved performance and safety. PMID- 15540749 TI - Comparison between extracorporeal shock wave lithotripsy and semirigid ureterorenoscope with holmium:YAG laser lithotripsy for treating large proximal ureteral stones. AB - PURPOSE: Management of large impacted upper ureteral calculi remains challenging for urologists. These calculi are frequently associated with obstructive uropathy and deteriorated renal function. Extracorporeal shock wave lithotripsy (SWL) is the least invasive treatment but its success rate is decreased for large impacted upper ureteral calculi. According to the American Urological Association guidelines on ureteral stones published in 1997, the appropriateness of ureteroscopy decreases when stone size exceeds 1 cm. However, the application of advanced ureteroscopy and techniques has increased the success rate of treating proximal ureter calculi. In this study we compared the safety and efficacy of ureterorenoscopic holmium:YAG laser lithotripsy (URSL) with SWL for large impacted proximal ureteral stones. MATERIALS AND METHODS: This study evaluated 82 patients with large impacted upper ureteral stones. Two patients were excluded from study owing to conversion to open surgery. SWL and URSL were performed in 41 and 39 patients, respectively. Those in the SWL group were treated on an outpatient basis with Medispec Econolith 2000 (Medispec, Germantown, Maryland) under intravenous sedation. The URSL was performed with a 6/7.5Fr semirigid tapered ureterorenoscope and holmium:YAG laser with the patient under spinal anesthesia on an inpatient basis. Successful outcome was defined as the patient being stone-free on radiography 1 month after treatment. Stone size, success rate, postoperative complications and cost were analyzed in each group. RESULTS: A total of 80 patients were enrolled in this study. Hematuria and flank pain were the most common complaints in each group. Mean stone size +/- SD was 1.28 +/- 0.04 cm in the SWL group and 1.51 +/- 0.05 cm in URSL group (p = 0.0009). Accessibility of the semirigid ureterorenoscope for impacted upper ureteral stones was 95.1% (39 of 41) and the stone-free rate achieved after 1 sitting was 92% (36 of 39). The initial stone-free rate of in situ SWL was 61% (25 of 41). Notably, the initial stone-free rate in the URSL group was better than that of the SWL group (p = 0.003). The efficiency quotient was 0.53 for URSL and 0.59 for SWL. The average cost in the URSL group appears to be lower than in the SWL group. Both groups were free of major complications. CONCLUSIONS: This study demonstrated that URSL achieved excellent results for upper ureteral calculi greater than 1 cm. Thus, this procedure should be considered first line therapy for large proximal ureteral stones. PMID- 15540750 TI - Bladder calculus resulting from the migration of an intrauterine contraceptive device. PMID- 15540751 TI - Testicular microlithiasis and its relation to testicular cancer on ultrasound findings of symptomatic men. AB - PURPOSE: We reviewed testicular microlithiasis (TM) on scrotal ultrasound in relation to the incidence of testicular neoplasm in males 17 to 45 years old with scrotal symptoms. MATERIALS AND METHODS: We reviewed the radiographic and medical records of males age 4 weeks to 84 years with symptomatic complaint of scrotal pain or swelling between September 1998 and April 2002. Subgroup analysis was performed on 160 male patients between 17 and 45 years old since they were at higher risk for testicular carcinoma. RESULTS: TM was found in 12 patients (8%) and concomitant testicular neoplasm was found in 4 (33%) symptomatic individuals age 17 to 45. In the 148 (93%) patients without testicular microlithiasis, concomitant testicular neoplasm was found in 2 individuals (2%). Statistical analysis revealed a relative risk of testicular neoplasm in the presence of TM to be 36.5 (CI 4.2-429.6) in our symptomatic population. Sensitivity and specificity were 0.67 and 0.95, respectively. CONCLUSIONS: Symptomatic patients with TM exhibited an incidence of testicular neoplasm greater than that of the general population. An algorithm for the surveillance of patients with TM should be developed. The fact that our patient population was symptomatic at referral preselected toward a higher incidence of TM and testicular neoplasm compared to accepted incidence in the general population. Further investigation will require a multicenter trial to generate an adequate patient pool due to the low incidence of TM and testicular malignancy. PMID- 15540752 TI - Comparative study of the Yang-Monti channel and appendix for continent diversion in the Mitrofanoff and Malone principles. AB - PURPOSE: Continent conduits for Mitrofanoff and Malone procedures are widely used for incontinence management. Because the appendix is not always suitable or available, Monti proposed a transversely retubularized short segment of ileum. We present our experience and compare outcomes according to the type of conduit. MATERIALS AND METHODS: In a retrospective study from 1988 to 2003 case notes were reviewed specifying conduit characteristics, underlying disease, overall surgical management of incontinence and subsequent complications. Comparison was based on followup, details of complications and treatments. RESULTS: In 32 men and 14 women with a total of 65 conduits (Mitrofanoff in 41 and Malone procedure in 24) the condition was spina bifida in 29, bladder exstrophy in 5 and sacral agenesia in 3. Other cases included various origins. Mean age at operation was 14.2 years (range 1 to 31). Mean followup was 5.3 years (range 1 to 14.7). Appendix was used in 23 cases and a Yang-Monti channel was performed in 18 (ileum in 7 and sigmoid in 11) for urinary diversion. The Malone procedure was performed using appendix in 11 cases and a Yang-Monti tube in 13. At followup 61 conduits (93.8%) were in use and 4 (6.2%) were abandoned. Complications were cutaneous stenosis, kink blockage and stomal leakage. No significant differences were observed between the types and purposes of the conduit. CONCLUSIONS: With constant availability and various possibilities of construction the Yang-Monti channel is recommended for continent diversion creation when appendix does not look suitable or is already in use or absent. PMID- 15540753 TI - Post-void residual urine volume in women with overactive bladder symptoms. AB - PURPOSE: Most patients with overactive bladder symptoms initially present to their primary care provider for treatment. It is currently recommended that post void residual urine (PVR) should be measured during the assessment of women complaining of overactive bladder symptoms and anticholinergic medication should be used if PVR is low. However, many primary care providers are reluctant to measure PVR and, therefore, they may delay treatment or even inappropriately treat patients who have a high PVR. We identified clinical factors that are associated with increased PVR that could be used to select which patients are at risk for this. MATERIALS AND METHODS: The charts of women presenting to our clinic with complaints of urinary frequency, urgency and/or urge incontinence were retrospectively reviewed. Patient demographics, physical examination and PVR were analyzed to determine if any factor was associated with increased PVR (100 ml or greater). RESULTS: The charts of 201 patients with complete data were reviewed. Patient age was 20 to 90 years (mean 55). Overall 19% of patients were found to have elevated PVR. Univariate analysis identified that older age, prior incontinence surgery, history of multiple sclerosis, greater American Urological Association Symptom Score, vaginal parity greater than 2, greater pad use and stage 2 or greater vaginal prolapse were associated with elevated PVR. Multivariate analysis identified age older than 55 years, prior incontinence surgery, a history of multiple sclerosis and vaginal prolapse stage 2 or greater as independent predictors of elevated PVR. CONCLUSIONS: It is possible to identify patients with overactive bladder symptoms who are at risk for elevated PVR based on history and physical examination. This could result in fewer patients needing PVR measurement prior to initiating treatment. PMID- 15540754 TI - Detrusor overactivity. Does it represent a difference if patients feel the involuntary contractions? AB - PURPOSE: We evaluated the differences between patients with overactive bladder (OAB) who felt involuntary detrusor contractions during cystometry (detrusor overactivity [DO]) and those who did not feel them. MATERIALS AND METHODS: We prospectively studied 45 patients with symptoms of nonneurogenic, nonobstructed overactive bladder and with DO on cystometry. All patients underwent videourodynamics, the ice water test and electrical perception threshold determination. Continence, urodynamic parameters, data from specific sensory evaluation and outcome of drug treatment were examined. RESULTS: Almost half of our patients did feel the contractions of DO and half did not. The groups differed significantly. Those without DO sensation were more frequently incontinent, had more involuntary detrusor contractions and these occurred earlier during bladder filling. They had involuntary start of voiding more frequently, more pathological sensation of bladder filling and lower electrical sensory thresholds. The results of drug treatment were better in the group who felt DO. CONCLUSIONS: Contractions of DO are felt by some of the patients and they differ from those patients who do not feel such contractions. It is likely that this finding reflects the existence of different OAB conditions with a different neuropathological cause and a different treatment outcome. Therefore, we suggest that specific tests for the evaluation of sensation in the lower urinary tract should be part of the diagnosis of patients with DO and symptoms of OAB. PMID- 15540755 TI - Randomized, double-blind placebo controlled trial of the once daily antimuscarinic agent solifenacin succinate in patients with overactive bladder. AB - PURPOSE: In this phase 3 trial we assessed the efficacy of solifenacin 5 mg and 10 mg daily in patients with symptoms related to overactive bladder. In addition, we assessed the safety and acceptability of solifenacin. MATERIALS AND METHODS: The study was a multicenter, multinational, randomized, double-blind, placebo controlled trial. Patients were randomized to 12-week once daily treatment with solifenacin 5 mg, solifenacin 10 mg or placebo. The primary efficacy variable was changed from baseline to study end point in mean number of micturitions per 24 hours. Secondary efficacy variables included changes from baseline in mean number of urgency, nocturia and incontinence episodes per 24 hours, and mean volume voided per micturition. RESULTS: Compared with changes obtained with placebo ( 1.59), micturitions per 24 hours were statistically significantly decreased with solifenacin 5 mg (-2.37, p = 0.0018) and solifenacin 10 mg (-2.81, p = 0.0001). A statistically significant decrease was observed in the number of incontinence episodes with both solifenacin doses (5 mg, p = 0.002 and 10 mg, p = 0.016). This effect was also seen for episodes of urge incontinence (5 mg, p = 0.014 and 10 mg, p = 0.042). Of patients reporting incontinence at baseline, fully 50% achieved continence after treatment with solifenacin. Episodes of nocturia were statistically significantly decreased in patients treated with solifenacin 10 mg (-0.71, -38.5%) versus placebo (-0.52, -16.4%, p = 0.036). Episodes of urgency were statistically significantly reduced with solifenacin 5 mg (-2.84, -51%, p = 0.003) and solifenacin 10 mg (-2.90, -52%, p = 0.002). Mean volume voided per micturition was statistically significantly increased with both solifenacin doses (p = 0.0001). Treatment with solifenacin was well tolerated. Dry mouth, mostly mild in severity, was reported in 7.7% of patients receiving solifenacin 5 mg and 23% receiving solifenacin 10 mg (vs 2.3% with placebo). CONCLUSIONS: In this study treatment with solifenacin 5 mg and 10 mg once daily significantly improved all the major symptoms of overactive bladder including frequency, urgency and incontinence. Solifenacin 10 mg also decreased the frequency of nocturia. Solifenacin therapy was associated with a favorable tolerability profile and a low incidence of dry mouth, especially at the 5 mg starting dose. PMID- 15540756 TI - Erosion of a male polypropylene sling. PMID- 15540757 TI - Holmium laser enucleation versus transurethral resection of the prostate: results from a 2-center, prospective, randomized trial in patients with obstructive benign prostatic hyperplasia. AB - PURPOSE: To our knowledge we report the first multicenter, prospective, randomized study comparing holmium laser enucleation (HoLEP) and transurethral prostate resection (TURP) for obstructive benign prostatic hyperplasia. MATERIALS AND METHODS: From January to October 2002, 100 consecutive patients with symptomatic obstructive benign prostatic hyperplasia were randomized at 2 centers to surgical treatment with HoLEP (52 in group 1) or TURP (48 in group 2). Patients in the 2 groups were preoperatively assessed by scoring subjective symptoms questionnaires. Preoperative and perioperative parameters were also evaluated, the latter at 1, 6 and 12 months of followup. RESULTS: At baseline all patients had obstruction (Schafer grade greater than 2). At the 1, 6 and 12-month followups no statistically significant differences were observed between the 2 groups in terms of urodynamic findings and subjective symptom scoring. In the HoLEP group mean total time in the operating room +/- SD was significantly longer than for TURP (74 +/- 19.5 vs 57 +/- 15 minutes, p < 0.05), while catheterization time (31 +/- 13 vs 57.78 +/- 17.5 minutes, p < 0.001 and hospital stay (59 +/- 19.9 vs 85.8 +/- 18.9 hours, p < 0.001) were significantly shorter in the HoLEP group. Transient stress and urge incontinence were more common in the HoLEP group, although at the 12-month followup results were comparable. The overall complication rate was comparable in the 2 groups. Erectile function was also maintained in the followup period from baseline in each group, as expected. CONCLUSIONS: HoLEP and TURP were equally effective for relieving obstruction and lower urinary tract symptoms. HoLEP was associated with shorter catheterization time and hospital stay. At 1 year of followup complications were similar in the 2 groups. PMID- 15540758 TI - Evaluation of autonomic innervation of the clitoris and bulb. AB - PURPOSE: Female sexual dysfunction is a common disorder but there are few objective measures of female genital function. We present a novel technique to record electrical activity of the female genital erectile tissue. MATERIALS AND METHOD: We placed concentric needle electrodes in the right clitoral crus and the bulb of 22 healthy women. Following spontaneous activity recording we stimulated the median nerve to activate the sympathetic nervous system. We simultaneously measured evoked activity in the clitoris and bulb, and hand sympathetic skin responses (SSRs). RESULTS: Spontaneous electrical activity of the clitoris and bulb was present in 13 subjects. A total of 21 subjects had evoked activity in the bulb, while 18 had evoked activity in the clitoris. All subjects had hand SSRs. Evoked genital response latencies were similar to hand SSR latency (p > 0.05). Evoked electrical activity was recordable in the clitoris and bulb but it was more easily performed in the bulb. CONCLUSIONS: Evoked electrical activity is more consistently obtained than spontaneous activity. These tests represent possible objective measures of female genital autonomic innervation, which may have relevance to female sexual function. PMID- 15540759 TI - Interaction between the phosphodiesterase 5 inhibitor, tadalafil and 2 alpha blockers, doxazosin and tamsulosin in healthy normotensive men. AB - PURPOSE: Tadalafil, a phosphodiesterase type 5 inhibitor, is effective therapy for erectile dysfunction (ED). Men with ED have a high incidence of comorbid conditions including cardiovascular disease, diabetes mellitus and benign prostatic hyperplasia. Although phosphodiesterase type 5 inhibitors are safe when administered with most medications, sildenafil given with doxazosin and vardenafil given with terazosin evoke orthostatic hypotension in some patients. We examined the hemodynamic interactions of tadalafil with the alpha-blockers doxazosin and tamsulosin. MATERIALS AND METHODS: In separate double-blind, placebo controlled, randomized crossover studies (18 patients in each study) we evaluated the hemodynamic effects of doxazosin 8 mg with tadalafil 20 mg, and tamsulosin 0.4 mg with tadalafil 10 and 20 mg. Blood pressure (BP) and heart rate were recorded before dosing and for 24 hours after dosing. RESULTS: Tadalafil 20 mg augmented the hypotensive effect of doxazosin by producing a mean maximal decrease in standing systolic BP (SBP) that was significantly greater than placebo (a mean difference of 9.8 mm Hg). Analysis of BP outliers showed that the number of subjects with a standing SBP of less than 85 mm Hg was greater after doxazosin plus tadalafil (28%) versus doxazosin plus placebo (6%). In subjects on tamsulosin, tadalafil 10 and 20 mg produced mean maximal decreases in standing SBP that were similar to placebo (mean difference of 1.7 and 2.3 mm Hg, respectively). No subject taking tamsulosin had a decrease in standing SBP less than 85 mm Hg. CONCLUSIONS: Tadalafil augmented the hypotensive effects of doxazosin but had little hemodynamic interaction with tamsulosin. In patients taking tadalafil for ED, tamsulosin 0.4 mg may be given for the treatment of benign prostatic hyperplasia. PMID- 15540760 TI - The presence of vasal vessels in men with congenital bilateral absence of the vas deferens. AB - PURPOSE: We characterized spermatic cord microanatomy in men with congenital bilateral absence of the vas deferens (CBAVD) presenting for varicocelectomy. We discuss the implications of these findings for varicocele repair. MATERIALS AND METHODS: Between 1997 and 2003, 8 men with CBAVD underwent a total of 11 microsurgical subinguinal varicocelectomies at microsurgical epididymal sperm aspiration and cryopreservation. All 8 men had palpable grades II to III varicoceles and in 6 varicoceles were repaired due to painful symptomatology, while 2 had testicular hypotrophy with an abnormal hormonal profile. Three men had bilateral varicoceles repaired, while 5 underwent unilateral varicocelectomy. All patients provided a thorough history and underwent physical examination, hormonal evaluation, semen analysis, genetic testing and renal ultrasonography. RESULTS: Intraoperative microsurgical dissection confirmed dilated internal and external spermatic veins, and absence of the vas deferens in all 11 spermatic cords. Characteristic tortuous vasal vessels of normal caliber were clearly identified in all 11 (100%) of these spermatic cords between the internal and external spermatic fasciae in the location where the vas deferens is usually found. CONCLUSIONS: Despite the absence of the vas deferens normal sized, orthotopically located vasal vessels were present in 100% of the spermatic cords examined. Furthermore, the caliber of the vasal veins was sufficient to provide adequate venous return from the testis following ligation of the internal and external spermatic veins. In patients with CBAVD presenting for varicocele repair standard microsurgical varicocelectomy with ligation of all internal and external spermatic veins can be performed without the risk of testicular congestion secondary to inadequate venous drainage. PMID- 15540762 TI - Laparoscopic mobilization of the retroperitoneal vas deferens for microscopic inguinal vasovasostomy. PMID- 15540761 TI - Prediction of successful outcome of microdissection testicular sperm extraction in men with idiopathic nonobstructive azoospermia. AB - PURPOSE: Microsurgical techniques in testicular sperm extraction can improve sperm retrieval in patients with nonobstructive azoospermia (NOA). However, spermatozoa retrieval rates have still been reported to be around 50% for patients with NOA. Thus, a reliable prediction method for successful outcome is needed to avoid unnecessary surgery. In this retrospective study we determined the diagnostic and predictive values of noninvasive parameters used in the treatment of patients with NOA. MATERIALS AND METHODS: We analyzed 9 preoperative factors including patient age, testicular volume and endocrinological data of 100 patients with NOA using multivariate logistic modeling. Testicular spermatozoa were retrieved successfully in 41 of the 100 patients (41%). RESULTS: We found that the concentrations of follicle-stimulating hormone (FSH), total testosterone (TT) and inhibin B were considered the most influential preoperative factors. We developed a formula to calculate the probability of successful outcome, P = [1 + exp(5.201 - 0.048 x FSH - 0.449 x TT - 0.021 x inhibin B)](-1). Association of predicted probabilities and observed responses was 0.77. A predicted probability of more than 15.7% was found to be the best cutoff. Sensitivity was 71.0% and specificity was 71.4% as determined by receiver operating characteristic analysis. CONCLUSIONS: We concluded that our formula should be useful for doctors considering microdissection testicular sperm extraction for patients with NOA because our equation uses noninvasive parameters without a preoperative testicular biopsy, which is a relatively invasive examination. PMID- 15540763 TI - A new approach to urology training: a laboratory model for percutaneous nephrolithotomy. AB - PURPOSE: The efficacy of traditional operating room based training of urology residents is being reevaluated. The development of hands-on laboratory practicums to facilitate the acquisition of skills by surgical residents lessens learning curves and hastens familiarity with tissue and instrument handling. We describe an innovative model for simulated percutaneous renal access and nephrolithotomy. MATERIALS AND METHODS: Porcine kidneys pre-implanted with artificial stone material were placed within intact chicken carcasses as a model for percutaneous nephrolithotomy. Urology residents were taught needle access, tract dilation and renal access sheath insertion using fluoroscopy. Training in percutaneous nephrolithotomy with the nephroscope, graspers and stone fragmentation methods followed. RESULTS: This simple, cost-effective model closely simulates percutaneous nephrolithotomy. Anonymous evaluations submitted by training session participants revealed a high degree of satisfaction with model effectiveness in the application of percutaneous renal access and nephrolithotomy techniques. CONCLUSIONS: Our percutaneous nephrolithotomy laboratory model is an effective means of skills acquisition for a complex endourological procedure. Patient care can safely be of secondary importance with respect to trainee experience in a low stress environment that provides an opportunity for supervised, repetitive performance of essential technical skills. We describe an effective percutaneous renal access and nephrolithotomy surgical training model of original design. PMID- 15540764 TI - Face, content and construct validity of the University of Washington virtual reality transurethral prostate resection trainer. AB - PURPOSE: We examined the face, content and construct validity of version 1.0 of the University of Washington transurethral prostate resection (TURP) trainer. MATERIALS AND METHODS: Version 1.0 of a virtual reality based simulator for transurethral skills was developed at our laboratory by integrating TURP hardware with our virtual 3-dimensional anatomy, irrigation control, cutting, bleeding and haptics force feedback. A total of 72 board certified urologists and 19 novices completed a pre-task questionnaire, viewed an introductory training video and performed a pre-compiled 5-minute resection task. The simulator logged operative errors, gm resected, blood loss, irrigant volume, foot pedal use and differential time spent with orientation, cutting or coagulation. Trainees and experts evaluated the simulator on a modified likert scale. The 2-tailed Levene t test was used to compare means between experts and novices. RESULTS: Overall version 1.0 content was between slightly and moderately acceptable. Experts spent less time with orientation (p < 0.0001), resected more total tissue (p < 0.0001), had more gm resected per cut (p = 0.002) and less blood loss per gm resected (p = 0.032), used less irrigant per gm resected (p = 0.02) and performed fewer errors (p < 0.0001) than novices. CONCLUSIONS: We established the face, content and construct validity for version 1.0 of the University of Washington TURP trainer to simulate the skills necessary to perform TURP. A predictive validity study showing a translation of skills from the virtual environment to the operating room will complete the validation of this model. PMID- 15540765 TI - The value of your time: evaluation of effects of changes in medicare reimbursement rates on the practice of urology. AB - PURPOSE: Several reforms to Medicare have changed the reimbursement of physicians from payment based on usual, customary or reasonable charges to a resource based relative value scale. We studied the effect of these changes on hourly reimbursement rates for various services provided by urologists. MATERIALS AND METHODS: We used a previously published national survey of urologists who provided information regarding physician time and work required before, during and after most frequently performed urological services, including during the global period. For comparison mean operative times during the last year at our private hospital for several common urological procedures were obtained. Medicare reimbursement rates for common urological procedures and evaluation and management (E&M) codes for 1995, 1999 and 2004 were acquired from our department's billing office and used to calculate reimbursement rate per hour. RESULTS: There was a steady increase in reimbursement for outpatient services and a decrease in reimbursement for surgical procedures. For E&M codes the reimbursement rates per hour for 2004 represent a mean 51% increase since 1995. However, surgical procedures have had a mean decrease of 28.5% in reimbursement rates per hour. There was remarkable consistency in rates with 7 of the 9 surgical procedures losing between 25.5% and 32% in reimbursement. In 1995 outpatient E&M services were the least profitable at less than half the hourly rate of operative procedures. In 2004 office cystoscopy and transrectal ultrasound biopsy of the prostate had the highest reimbursement and, with the exception of shock wave lithotripsy, there was a minimal difference in hourly reimbursement rates between common surgical procedures and E&M services. CONCLUSIONS: Changes in Medicare reimbursement during the last decade have resulted in significant changes in rates for different urological services. The near equity in reimbursement rates for E&M and surgical services will likely have an increasingly important role in the future practice of urology. PMID- 15540766 TI - Bladder calculi after augmentation cystoplasty: risk factors and prevention strategies. AB - PURPOSE: Lower urinary tract reconstruction is an essential tool in the management of severely dysfunctional bladders in children. The incidence of calculi in augmented bladders has been reported in up to 50% of cases. We analyzed our experience with stone formation in this population to assess risk factors and outcomes. MATERIAL AND METHODS: We performed a retrospective cohort study of all patients who underwent bladder augmentation from 1988 to 2002 at our institution. Patient demographics, risk factors and management were abstracted from the medical record. RESULTS: A total of 105 patients (58 males and 47 females) were identified. Ileum, colon and stomach were used in 37, 18 and 50 patients, respectively. Median age was 8.0 years. Median followup was 8.4 years. A total of 12 patients (11%) were found to have bladder calculi. Ten patients with ileum (27%), 1 with colon (6%) and 1 with stomach (2%) formed stones. All patients had recurrent urinary tract infections. Nine patients were successfully treated with an endoscopic procedure. Four patients (33%) formed recurrent stones despite saline bladder irrigations. One patient had multiple recurrences but is now stone-free on a daily regimen of 20% urea instillation. CONCLUSIONS: Augmentation cystoplasty carries an overall low risk of bladder calculi. Gastrocystoplasty had a significantly lower rate of stone formation than augmentation with ileum and colon. Urinary tract infection is an independent risk factor for stone formation. Endoscopic management is safe and effective in the majority of patients and it may be facilitated by a percutaneous access. Recurrent stones form in some patients despite aggressive medical management. PMID- 15540767 TI - The use of stab incisions for instrument access in laparoscopic urological procedures. AB - PURPOSE: Traditional laparoscopic procedures use expensive cannulas to facilitate the insertion and removal of laparoscopic instruments. We report our experience with limited access stab incisions for the insertion of instruments into the peritoneal cavity during laparoscopic urological procedures to minimize the use of disposable cannulas. MATERIALS AND METHODS: All patients undergoing laparoscopic urologic procedures using stab incisions, as performed by us, from November 1999 through March 2003 were included. Procedures included nephrectomy, partial nephrectomy, varicocelectomy, nephroureterectomy, orchiopexy and adrenal procedures. A single cannula was used for telescope access. In select cases additional cannulas were used for unique instruments or specimen manipulation/extraction. Abdominal wall stab incisions were used for the remaining instruments. Stab incisions were closed with a Steri-Strip (3M Healthcare, St. Paul, Minnesota) at the skin level only. RESULTS: A total of 53 procedures were performed during the study period. Pneumoperitoneum was maintained in all cases. There were no complications associated with the use of stab incisions. A total of 105 cannulas were saved using our technique. At a cost to the patient of dollars 140 per cannula the overall cost saving was dollars 14,700 with an average saving of dollars 277 per case. CONCLUSIONS: Laparoscopic urological procedures can be performed effectively and safely with stab incisions for instrument access. There are significant cost savings related to the elimination of cannulas. We believe that our technique of stab incisions for instrument access is equivalent to the traditional cannula approach and should be used when possible. PMID- 15540768 TI - Long-term followup of buccal mucosa onlay graft for hypospadias repair: analysis of complications. AB - PURPOSE: We review the long-term results of buccal mucosa onlay grafting for urethral reconstruction in hypospadias surgery in patients with followup of at least 5 years. MATERIALS AND METHODS: We retrospectively evaluated 132 patients who underwent buccal mucosa onlay graft for hypospadias repair, including 34 who underwent "salvage" grafting, during a 10-year period at our institution. In 49 cases with available followup longer than 5 years (average 6.2) complications were analyzed in detail. RESULTS: The overall complication rate was 24% (12 of 49 patients), with all but 3 complications occurring during the first postoperative year (3 fistulas, 1 stricture, 1 meatal stenosis, 2 graft contractures, 2 scars at oral harvesting site). The remaining 3 complications manifested during postoperative years 2 and 3, and encompassed 2 anastomotic strictures at the proximal anastomosis and 1 meatal stenosis. CONCLUSIONS: Buccal mucosa onlay grafts for urethral reconstruction in hypospadias repair seem to provide stable long-term results, with complications occurring primarily during the first 12 months postoperatively. PMID- 15540769 TI - Comparison of hypospadiac and normal preputial vascular anatomy. AB - PURPOSE: Data about the differences between the vascularization of normal and hypospadiac prepuce are lacking. We investigated the course of the preputial arterial blood vessels in normal controls and children with hypospadias by using transillumination, arterial methylene blue injection and 3-dimensional (3-D) reconstruction of serial histological sections focusing on arterial vessels. MATERIALS AND METHODS: Prepuce of 48 normal controls and 15 children with hypospadias was transilluminated by a front and back lighting technique and then photographed. All of the normal and 12 of hypospadiac prepuces not used for urethroplasty or penile body skin reconstruction were removed. The blood vessels of normal prepuce were also identified after arterial injection of methylene blue. Selected prepuce of normal controls and children with hypospadias was serially sectioned, and arterial and venous vessels were histologically distinguished. A 3-D computer reconstruction of the arterial system of normal and hypospadiac prepuces was performed. RESULTS: We confirmed the reliability of the transillumination technique to describe the arterial vascular anatomy of the prepuce by comparing the transillumination to methylene blue injection and 3-D reconstruction of histological sections. We classified the arterial vascular anatomy of normal prepuce as 1 artery predominant (41.67%), 2 arteries predominant (25%), H-type arching artery (12.5%) and net-like arterial system (20.83%). However, hypospadiac prepuce revealed a net-like arterial system more frequently (50%). We noted that the frequency of net-like arterial system was higher in more severe hypospadiac prepuce. CONCLUSIONS: Understanding the differences between normal and hypospadiac prepuce vascular anatomy is germane to hypospadias surgery. The arterial blood supply of the hypospadiac prepuce is different than normal. A better knowledge of the vascular anatomy of the hypospadiac prepuce may improve the surgical results of hypospadias repair. PMID- 15540771 TI - Ability of a normal dysfunctional voiding symptom score to predict uroflowmetry and external urinary sphincter electromyography patterns in children. AB - PURPOSE: We correlated uroflowmetry/electromyography (EMG) patterns and post-void residual urine volume (PVR) in children with a normal dysfunctional voiding symptom score (DVSS) and determined whether a normal DVSS can predict uroflowmetry/EMG/PVR results. MATERIALS AND METHODS: We prospectively studied 51 children with a normal DVSS, and negative neurological and urological history who underwent uroflowmetry/EMG and PVR measurements. Statistical results were correlated with patient age and gender using Cohen's kappa and Fisher's exact test. RESULTS: All children had a normal DVSS for gender and 37 (73%) had bell shaped uroflowmetry curves. Eight of the 14 children with nonbell-shaped uroflowmetry curves had plateaued and 6 had fractionated curves. Five boys and 13 girls (35%) had decreased EMG patterns, 7 boys and 8 girls (29%) had increased patterns and 5 boys and 13 girls (35%) had variable patterns (Cohen's kappa score 0.11). Of the 37 children (65%) with bell-shaped uroflowmetry curves 24 had normal PVR, while 13 of 37 (35%) had elevated PVR. Eight of the 14 children (57%) with nonbell-shaped uroflowmetry curves had normal PVR and 6 (43%) had elevated PVR (Cohen's kappa score 0.13). Seven of 37 children (19%) with bell-shaped uroflowmetry curves and 7 of 14 (50%) with nonbell-shaped uroflowmetry curves had a significantly distended bladder (p = 0.04). Nine of the 14 children (64%) with a significantly distended bladder had abnormal PVR. Ten of the 37 children (27%) without a significantly distended bladder had abnormal PVR (p = 0.02). CONCLUSIONS: Normal DVSS for gender is able to predict a bell-shaped uroflowmetry curve in most children, while EMG patterns and PVR do not correlate with uroflowmetry curves in these children. Children with a significantly distended bladder frequently have nonbell-shaped uroflowmetry curves as well as elevated PVR. PMID- 15540770 TI - Giant bladder diverticula causing bladder outlet obstruction in children. AB - PURPOSE: Congenital giant diverticulum of the bladder is a consequence of deficiency in the detrusor musculature and has been reported in male infants as a rare cause of bladder outlet obstruction. MATERIALS AND METHODS: A 10-year retrospective review revealed 4 patients (3 boys and 1 girl) with bladder outlet obstruction due to a giant bladder diverticulum. Prenatal and postnatal clinical and imaging records were reviewed. RESULTS: Prenatal sonography was unremarkable in all patients. Three males (ages 4 months, 10 months and 3 years) had no medical history of voiding dysfunction, and 1 female (11 years) had the Ehlers Danlos syndrome. While the girl presented with urinary tract infection, all patients presented with progressively decreasing urinary stream and urinary retention. Each patient underwent voiding cystourethrography (VCUG) and ultrasound. In each patient VCUG showed a giant (greater than 7 cm) bladder diverticulum that descended below the bladder neck and compressed the urethra during voiding. Vesicoureteral reflux was seen in 2 patients. Ultrasonography demonstrated moderate unilateral hydronephrosis in 2 patients. Bladder diverticulectomy was successfully performed in all patients, with ureteral reimplantation in 3. CONCLUSIONS: A giant congenital bladder diverticulum, when noted on VCUG to descend below the bladder neck, may lead to bladder outlet obstruction. To our knowledge we report the first case of a female presenting with bladder outlet obstruction due to a giant bladder diverticulum. Children with connective tissue disorders may be predisposed to this disorder, which must be excluded, regardless of gender, in all patients presenting with voiding abnormalities. Surgical diverticulectomy, often with ureteral reimplantation, is the preferred treatment, with excellent long-term results. PMID- 15540772 TI - The diameter of the rectum on ultrasonography as a diagnostic tool for constipation in children with dysfunctional voiding. AB - PURPOSE: We proved the accuracy of the transverse diameter of the rectum on ultrasonography as an additional parameter for diagnosing constipation in children with lower urinary tract dysfunction. MATERIALS AND METHODS: The diameter of the rectum on bladder ultrasonography in a constipated group of patients with dysfunctional voiding was compared to this diameter in a control group of patients with a normal defecation pattern. A total of 49 children were included. Group 1 consisted of 23 patients with a positive history of dysfunctional voiding and, according to pediatric gastroenterological practice, constipation. Control group 2 consisted of 26 patients without lower urinary tract dysfunction and a normal defecation pattern. In each group a defecation questionnaire was administered and physical examination of the abdomen was done. In all patients a 7.5 MHz probe was used to measure the transverse diameter of the rectum behind the bladder on ultrasonography. The probe was applied on the abdominal skin approximately 2 cm above the symphysis. Measurement was performed with a filled bladder at an angle of about 15 degrees downward from the transverse plane. RESULTS: In constipated group 1 the mean diameter of the rectum was 4.9 cm (95% CI 4.4 to 5.3). In the control group the mean diameter of the rectum was 2.1 cm (95% CI 1.8 to 2.4). In group 1 the diameter of the rectum was significantly larger than in group 2 (p < 0.001). None of the patients had a sensation to defecate during the investigation. There was no significant difference in age between the 2 groups (p = 0.20) and no significant difference between them in the period between the last time that stool was passed prior to the time of rectal measurement (p = 0.16). CONCLUSIONS: The transverse diameter of the rectum measured by lower abdominal ultrasound provides an additional accurate parameter with which to diagnose constipation in patients with nonneurogenic bladder-sphincter dyssynergia. PMID- 15540773 TI - Heterozygotic twinning in a case of female vesicourethral duplication. PMID- 15540774 TI - Age and lean body weight related growth curves of kidneys using real-time 3 dimensional ultrasound in pediatric urology. AB - PURPOSE: Kidney volume and growth are important parameters for evaluating and monitoring several diseases in pediatric urology. Studies have shown the significant relationship of kidney volume and function. Until now normal values of kidney size and growth have been based on 2-dimensional measurements combined with arithmetic formulas for the ellipsoid. Real-time 3-dimensional (RT3D) ultrasound is a new imaging modality. Moreover, corresponding software allows RT3D imaging within a short time exposure. We created nomograms of kidney volumes for routine diagnostics that could serve as a standard for pediatric renal growth assessment, decreasing the need for invasive tests. MATERIALS AND METHODS: RT3D volumetry of a total of 620 kidneys in 310 children with a homogeneous age distribution (range newborn to 10 years) and without any evidence of renal impairment was performed using a Voluson 730 (Kretztechnik, Ultrasound, Zipf, Austria) system. Polynomial regression analysis was applied for the prediction and estimation of growth variables of kidney volumes as a function of gender, age, body mass index or lean body weight. RESULTS: Stepwise multiple regression analysis incorporating the different independent variables indicated with significant correlation that age and lean body weight were the strongest predictors of kidney volumes in children. CONCLUSIONS: This study shows that RT3D ultrasound is particularly qualified for evaluating kidney volume in pediatric urology. RT3D volumetry is suitable for monitoring renal growth, reflecting kidney function. Furthermore, this methodology is feasible in screening programs assessing congenital urogenital diseases. PMID- 15540775 TI - Fetal bladder sagittal length: a simple monitor to assess normal and enlarged fetal bladder size, and forecast clinical outcome. AB - PURPOSE: We examined if the parameter of fetal bladder sagittal length (FBSL) could serve as a monitor of normative and enlarged fetal bladder size. MATERIALS AND METHODS: There were 76 consecutive cases examined between 1984 and 2000 that included measurement of fetal bladder size as FBSL and postnatal urological followup. Fetal images used to assess normal bladder size were derived from cases in which the bladder was normal on prenatal imaging and postnatal testing. An enlarged bladder was categorized as being greater than the 95% CI for a given gestational age (GA). The presence and extent of renal pelvic dilatation were also noted and correlated with FBSL. RESULTS: Measuring normal FBSL in 39 fetuses showed an exponential growth pattern (r = 0.76), which could be represented by the approximate linear formula FBSL = GA in weeks -5 (+/- 95% upper/lower CI = 7). An enlarged bladder was diagnosed in 37 fetuses. A dilated bladder in 9 fetuses, defined as FBSL greater than the 95% upper CI of normal (ie between GA + 2 and GA + 12), showed outcomes of posterior urethral valves, vesicoureteral reflux or a normal outcome. Megacystis in 28 fetuses, defined as FBSL greater than 10 mm larger than that of a dilated bladder (ie greater than GA + 12), showed additional outcomes of megacystis megaureter/vesicoureteral reflux or prune-belly syndrome. A normal outcome was significantly more likely in fetuses with a dilated bladder than in those with megacystis (p < or = 0.05). The incidence of azotemia in those with a dilated bladder or megacystis and pyelectasis was significantly lower than that in those with megacystis with hydronephrosis (p < or = 0.03). CONCLUSIONS: Postnatal diagnosis of fetuses that show an enlarged bladder is predicted based on whether the bladder is enlarged as a dilated bladder or megacystis and if the renal pelvis is enlarged as pyelectasis or hydronephrosis. PMID- 15540776 TI - Unilateral nephrectomy as palliative therapy in an infant with autosomal recessive polycystic kidney disease. AB - Prenatal diagnosis of autosomal recessive polycystic kidney disease (ARPKD) implies a dire prognosis. Neonates affected by the more severe variants of ARPKD suffer respiratory failure caused by massive kidneys that restrict diaphragmatic expansion and result in pulmonary hypoplasia. Afflicted infants who survive the neonatal period and gain adequate respiratory function may subsequently suffer from an inability to tolerate enteral nutrition due to abdominal compression from the massive kidney and the systemic effects of renal compromise. Palliative unilateral or bilateral nephrectomy may be considered in rare instances to facilitate pulmonary expansion and gastrointestinal function. We report on an infant with severe ARPKD who was able to tolerate enteral nutrition only after left nephrectomy. PMID- 15540777 TI - Mullerian lesions of the bladder. PMID- 15540778 TI - Prostatic abscess. PMID- 15540779 TI - Comparison of laparoscopic saline infused to dry radio frequency ablation of renal tissue: evolution of histological infarct in the porcine model. AB - PURPOSE: We investigated and compared the gross and histopathological features of radio frequency (RF) ablated renal tissue using saline infused RF ablation (RFA) vs dry RFA in a porcine model. MATERIALS AND METHODS: Ten porcine kidneys underwent laparoscopic RFA. The lower and upper poles of each right kidney were treated with 1 and 2 cycles of saline augmented (wet) RFA, respectively. The upper pole of each left kidney was RF ablated without saline infusion (dry RFA) and a control lesion was created in the lower pole. The animals were sacrificed immediately after ', 2 and 9 days after (subacute), and 14 and 21 days after (chronic) treatment, respectively. Gross pathological and histological changes caused by the different RF treatments were compared. RESULTS: Mean time to attain target temperature was shorter with wet than with dry RFA. Grossly the lesion sizes achieved were larger using wet RFA but independent of the number of wet RFA treatment cycles. Gross lesion sizes decreased with time for each treatment modality. No histopathological differences were seen between the 2 RFA treatment modalities as well as between 1 vs 2 cycles of wet RFA. In the acute phase hematoxylin and eosin staining of ablated tissue revealed focal areas of alterations in renal tubular histology. However, nicotinamide adenine dinucleotide staining of corresponding areas confirmed the lack of cellular viability except for glomeruli. In the subacute phase there were focal coagulation necrosis and thrombosed blood vessels. Within the necrotic areas the glomeruli were the last structures to lose viability. In the chronic phase fibrosis with decreased lesion size was seen. Nicotinamide adenine dinucleotide staining demonstrated that the cellular kill was permanent during the entire study period. CONCLUSIONS: The effects of RFA of renal tissue has 2 phases, namely initial direct thermal ablation causing acute cell death and a later subacute phase causing subsequent infarction of the distal arterial vascular supply to the ablated region, resulting in more cell death and coagulative necrosis. Glomeruli are the last structures to lose viability in necrotic areas. Most importantly cellular death caused by RFA is permanent. PMID- 15540780 TI - Kinetics, biodistribution and therapeutic efficacy of hexylester 5 aminolevulinate induced photodynamic therapy in an orthotopic rat bladder tumor model. AB - PURPOSE: To optimize photodynamic therapy (PDT) we investigated the kinetics and biodistribution of hexylester 5-aminolevulinate (hALA) induced protoporphyrin IX (PpIX) and the therapeutic efficacy of PDT at different drug and light doses in an orthotopic rat bladder tumor model. MATERIALS AND METHODS: Healthy and tumor bearing rats were instilled intravesically with hALA (4, 8 and 16 mM) for 1 hour. Fluorescence was recorded spectroscopically in situ. PpIX fluorescence distribution and quantification across the bladders was visualized with fluorescence microscopy. PDT efficacy at different fluences (15 to 80 J/cm2) was histologically assessed 48 hours and 1 week after treatment. RESULTS: Spectroscopic analysis in normal or tumor bearing rats showed the highest tumor to-normal ratios 2 or 3 hours after the end of the 8 or 16 mM hALA instillation (5.4 and 5.7, respectively). Within the same tumor bearing animal the same fluorescence levels were observed in normal epithelium and transitional cell carcinoma, whereas the tumor-to-muscle ratio was 3. Tumor necrosis with an intact normal bladder wall was observed with a fluence of 20 J/cm2 for 8 mM hALA, while 15 J/cm2 was ineffective and 25 J/cm2 induced total wall necrosis. Although it induced comparable PpIX fluorescence, 16 mM hALA did not result in tumor eradication at any fluence. CONCLUSIONS: An optimal PDT effect was obtained with 8 mM hALA and a fluence of 20 J/cm2. While different hALA concentrations ind uce identical PpIX fluorescence intensities, the PDT outcome was considerably different. Thus, fluorescence does not necessarily predict the therapeutic efficacy of PDT. PMID- 15540781 TI - A novel intravesical therapy for superficial bladder cancer in an orthotopic model: oncolytic reovirus therapy. AB - PURPOSE: To our knowledge this is the first report of intravesical oncolytic reovirus for therapy of superficial bladder cancer in an orthotopic bladder tumor model. Superficial bladder carcinomas are often multifocal and have high recurrences after surgical resection. In 20% of cases intravesical immunotherapy fails to prevent recurrence and complications from bacillus Calmette-Guerin (BCG) are common. Human reovirus is an oncolytic virus that selectively destroys cancer cells with an activated Ras pathway. We examined the ability of this virus to kill bladder cancer cells in vitro and inhibit tumor growth in vivo. MATERIALS AND METHODS: Following cytotoxicity assays in vitro dose escalation of intravesical reovirus was tested for tumor control and toxicity in a rat model. Treatments were done twice weekly for 3 weeks. In parallel intravesical BCG was studied. Animals were monitored on a daily basis for health status and by routine urine cytology. Animals underwent necropsy at study end point and appropriate tissues were taken for histology. RESULTS: Side effects in reovirus groups were minor compared with BCG complications. Tumor response (animal survival) was 90% 100 days after tumor implantation in reovirus treated animals, whereas the highest survival in BCG treated groups was 50%. Animals treated with reovirus had significantly higher tumor-free survival than those treated with immunotherapy or normal saline (log rank test p = 0.0002 and 0.04, respectively). CONCLUSIONS: Intravesical reovirus is safe and effective in this animal model and it may have clinical applications for bladder cancer treatment. PMID- 15540783 TI - Identifying the combination of the transcriptional regulatory sequences on prostate specific antigen and human glandular kallikrein genes. AB - PURPOSE: The combination of prostate specific antigen (PSA) and human glandular kallikrein (KLK2) promoters and/or enhancers was used to establish a new model to determine the feasibility of tissue specific expression for prostate cancer. MATERIALS AND METHODS: In vitro studies used the construction of PSA and KLK2 promoters/enhancers vectors to elucidate the link between the promoter/enhancer of PSA and KLK2. Reverse transcriptase-polymerase chain reaction assays were used to determine cell specific expression. Therefore, an attractive tissue specific expression vector for PSA and KLK2 gene was identified. RESULTS: The reporter vectors driven by KLK2 promoter had much lower luciferase activities than those of the reporter vectors driven by PSA promoter in LNCaP cells. Furthermore, the most efficient and cell specific reporter activity after 5alpha-androstan-17beta ol-3-one treatment among the reporter vectors constructed in this study was that of pKLK2EPSABHE, which was driven by KLK2 enhancer and PSA promoter/enhancer. The pKLK2EPSABHE reporter vector could induce 800-fold higher than the KLK2 basic promoter and its reporter activity was 16 times that of the enhancer/promoter element of KLK2 following induction by androgen. CONCLUSIONS: The results verify that the PSA promoter/enhancer must be combined with KLK2 to ensure the full activity and cell specificity of the gene. These expressions coupled with mechanic target validation yield valuable clues regarding the model of action of complex mixtures. This model is a potentially useful tool in gene therapy for metastatic prostate cancer. PMID- 15540782 TI - Chemosensitization of bladder cancer cell lines by human telomerase reverse transcriptase antisense treatment. AB - PURPOSE: Responses of transitional cell carcinoma of the bladder (TCC) to commonly used chemotherapy agents such as mitomycin C (MMC), cisplatin and gemcitabine are often disappointing. Since human telomerase reverse transcriptase (hTERT) is tumor specifically expressed and contributes to the immortality and malignancy of the majority of tumors, it is regarded as a suitable antitumor target. We investigated whether combinations of hTERT antisense (AS) oligonucleotides (ODNs) with common chemotherapy (CT) schedules may improve drug mediated antitumor effects. MATERIALS AND METHODS: Initial screening for enhancement of the inhibitory effects of MMC, cisplatin and gemcitabine on viability by treatment with the 2 hTERT AS-ODNs ASt2206 and ASt2331 was performed in 4 TCC cell lines prior to CT. Apoptosis was assessed by annexin V staining and detection of activated caspase-3 using Western blot analysis. Nonsense (NS)-ODN served as a control in all experiments. RESULTS: All cell lines responded to the anticancer agents tested. Treatment with AS plus CT resulted in a significantly stronger inhibition of viability than the NS plus CT control in the majority of combinations, indicating an AS specific enhancement effect. For example, ASt2331 plus MMC decreased viability to 17% in contrast to NS plus MMC (58%) in EJ28 cells. All ASt2331 plus CT combinations specifically increased the rate of apoptosis 1.3 to 3.0-fold compared with NS plus CT. Apoptosis induction was associated with caspase 3 activation. CONCLUSIONS: Enhancement of cytotoxic drug effects on the growth of TCC cells by hTERT AS-ODNs presented herein allows a dose decrease in chemotherapy and confirms the suitability of hTERT as a target in a specific therapy approach. PMID- 15540784 TI - Down-regulation of the prostate specific antigen promoter by p53 in human prostate cancer cells. AB - PURPOSE: The WT p53 gene appears to have a broad role in suppressing prostatic tumorigenesis. We identified the mechanisms responsible for the effect of p53 on prostate specific antigen (PSA) expression by prostate cancer cell lines in vitro and investigated the role of a putative p53 response element in the PSA promoter region in prostate cancer cells. MATERIALS AND METHODS: LNCaP cells were used to determine the effect of doxorubicin on p53 and PSA expression. The putative p53 response element in the human PSA promoter was identified by transient gene expression with site direct mutagenesis assays using a PSA reporter vector. Quantitative PSA secretions were assessed using enzyme-linked immunosorbent assays. RESULTS: Enzyme-linked immunosorbent and immunoblot assays indicated that doxorubicin treatment increased p53 expression but inhibited PSA levels in LNCaP cells. Transient gene expression assays showed that human PSA promoter activity was blocked by doxorubicin treatment. Mutation of the p53 response element GGGCATGTCT to GGGAGGATCT abolished the blocking effects of doxorubicin on PSA gene promoter activity. CONCLUSIONS: Results demonstrate that p53 regulates PSA gene expression through a putative p53 response element in the PSA promoter within human prostate cancer cells. PMID- 15540785 TI - Genitourinary complications of inherited epidermolysis bullosa: experience of the national epidermylosis bullosa registry and review of the literature. AB - PURPOSE: We determined the frequency with which 6 genitourinary tract complications (urethral meatal stenosis, urinary retention, bladder hypertrophy, hydronephrosis secondary to ureteral strictures, pyelonephritis and cystitis) occur in inherited epidermolysis bullosa (EB) in the American EB population. MATERIALS AND METHODS: Up to 16 years of longitudinal followup was done in 3,280 consecutively enrolled patients in the National EB Registry, a National Institutes of Health funded epidemiological study covering the entire continental United States. Data were stratified by major EB type and subtype. Frequencies of occurrence were determined for each of 6 genitourinary tract variables and stratified into 10 mutually exclusive, major EB subtypes. RESULTS: Urinary tract complications occurred in a minority of patients across all major EB subtypes with the highest frequency seen in Herlitz junctional EB (JEB-H). Urethral meatus stenosis was the most common complication, occurring in 11.6% and 8.0% of patients with JEB-H and Hallopeau-Siemens recessive dystrophic EB (RDEB), respectively. Urinary retention, hydronephrosis and bladder hypertrophy occurred in 9.3%, 7.0% and 4.6% of JEB-H cases, respectively. In contrast, pyelonephritis and cystitis were most often seen in the setting of generalized EB simplex (Koebner variant) and inversa RDEB. CONCLUSIONS: The urinary tract may be involved in any subtype of inherited EB, although these complications usually arise in patients with the most severe subtypes of junctional and recessive dystrophic disease. Chronic surveillance for the presence of genitourinary tract disease activity is warranted, especially in patients with JEB and RDEB, given the potential for longterm kidney injury if untreated. PMID- 15540786 TI - Absorption of intravesically applied drugs: comparison of normal and ileal augmented rabbit bladder. AB - PURPOSE: Drug absorption within urinary diversions has been reported to cause prolonged and higher grade toxicity. Therefore, continuous urine drainage has been recommended during chemotherapy in patients with continent urinary diversion. We developed an animal model in which to examine the significance of drug absorption in normal rabbit bladders compared with ileal augmented bladders. MATERIALS AND METHODS: Ten rabbits with ileal bladder augmentation and 5 control animals were used for absorption studies with methotrexate and ofloxacin. One, 4 and 12 months after surgery the rabbits received an intravesical instillation of either drug. During 2 hours blood samples were drawn. To avoid overfilling the bladder by urine it was emptied after 30, 60, 90 and 120 minutes, and refilled with fresh solution to yield a relatively constant drug amount. After 12 months the animals were sacrificed. Area of the native bladder wall and of the intestinal segment was measured to allow the correction of absorbed drug amounts per surface area. Moreover, the median villous heights of native and augmented ileum were compared histologically. Serum levels of methotrexate and ofloxacin were determined by high performance liquid chromatography. RESULTS: The maximum serum concentration of the 2 drugs was typically seen after 60 to 120 minutes. During the 1-year period peak serum concentrations of ofloxacin remained consistently higher in ileal augmented than in control rabbits, although this did not achieve statistical significance at all instillation time points. For methotrexate a statistically significant difference was not shown for either time point. When absorption was corrected for total bladder surface area, the enhancement of ofloxacin absorption by ileal augmentation weakened and attained statistical significance only at the 1-month time point. Histological examinations after 12 months showed that augmented intestinal mucosa had a significantly smaller villous height than native ileum. CONCLUSIONS: Our data demonstrate that bladder surface is the most important factor for increased absorption but time dependent histological changes of the integrated intestinal mucosa also influence absorption. There is a broad interindividual discrepancy. Therefore, general rules in patients with urinary diversion may not be justified. PMID- 15540787 TI - Inhibition of the bladder cooling reflex in the awake state: an experimental study in the cat. AB - PURPOSE: We assessed the bladder cooling reflex in the awake cat. The bladder cooling reflex is consistently observed in anesthetized adult cats but not in awake, neurologically normal humans. This discrepancy could indicate a state dependant control of the reflex or a species difference. This study was designed to differentiate between these alternatives, MATERIALS AND METHODS: Under ketamine-xylazine 5 animals had an indwelling catheter inserted into the bladder. The cooling reflex was tested by injections of cold saline into the bladder (4C to 8C), lowering its wall temperature to about 30C to 32C. The volume used (5 ml) was subthreshold for the Adelta micturition reflex, as confirmed by control injections of body warm saline. The procedure was repeated with the animals fully awake and it was well tolerated by all of them. Reflex responses were assessed by induced bladder pressures. RESULTS: Typical bladder cooling reflexes with peak pressures greater than 3 kPa were evoked in all cats when in narcotic sleep (group mean +/- CI 7.4 +/- 3.1 kPa). No such reflexes were elicited when the animals were awake (2.0 +/- 1.0 kPa). The difference was significant at the level of individual animals. CONCLUSIONS: The bladder cooling reflex is suppressed in adult cats during wakefulness, as in humans. This state dependent control of the bladder cooling reflex adds to its resemblance to the extensor plantar response (Babinski's sign). PMID- 15540788 TI - Urodynamic effects of the K+ channel (KCNQ) opener retigabine in freely moving, conscious rats. AB - PURPOSE: Retigabine is a novel anticonvulsant drug that not only augments gamma aminobutyric acid mechanisms, but also opens voltage gated K+ channels (KCNQ). In this study we investigated the effects of retigabine on detrusor activity in rats. MATERIALS AND METHODS: To conscious, female Sprague-Dawley rats undergoing continuous cystometry retigabine was given intravenously (0.5, 1 and 5 mg/kg( 1)). The KCNQ channel blocker linopirdine was given intravenously (2 mg/kg(-1)) 5 minutes prior to retigabine (1 mg/kg(-1)). In addition, retigabine was given intracerebroventricularly (1, 5 and 10 microg) and intravesically (100, 500 and 1,000 ng ml(-1)). The effects of the drug (intravesical administration) on capsaicin induced bladder overactivity were also tested. RESULTS: Retigabine given intravenously (1 mg/kg(-1)) decreased baseline and maximal bladder pressures, increased voided and infused volumes, and increased voiding intervals. Retigabine (10 microg) given intracerebroventricularly decreased baseline pressure and increased voided and infused volumes as well as voiding intervals. However, bladder pressures were not significantly affected. Intravesical retigabine (1,000 ng.ml(-1)) decreased maximal bladder pressure, increased voided and infused volumes, and increased voiding intervals. Given intravesically for 30 minutes prior to intravesical capsaicin (30 microM) instillation retigabine (1,000 ng.ml(-1)) decreased the detrusor overactivity induced by capsaicin. The KNCQ channel blocker linopirdine (2 mg/kg(-1)) completely blocked the effects of intravenous retigabine (1 mg/kg(-1)). CONCLUSIONS: Retigabine given intravenously, intracerebroventricularly and intravesically increased micturition volume and voiding intervals and, when given intravesically, it decreased capsaicin induced detrusor overactivity, suggesting that KCNQ channels can be interesting targets for drugs aiming at micturition control. Retigabine may be a candidate to test as a treatment for detrusor overactivity in humans. PMID- 15540789 TI - Demonstration of bladder selective muscarinic receptor binding by intravesical oxybutynin to treat overactive bladder. AB - PURPOSE: The current study was done to elucidate the in vivo mechanism of action of intravesical instillation of oxybutynin to treat overactive bladder. MATERIALS AND METHODS: In rats receiving oral and intravesical oxybutynin we measured muscarinic receptors in the bladder and other tissues by radioligand binding assay using [3H]NMS ([N-methyl-3H] scopolamine methyl chloride) with the simultaneous measurement of plasma concentrations of oxybutynin and its active metabolite N-desethyl-oxybutynin. Pilocarpine induced salivary secretion was also measured. RESULTS: Following oral administration of oxybutynin there was a significant increase in the apparent dissociation constant (Kd) for specific [3H]NMS binding in the bladder, submaxillary gland, heart and colon of rats at 1 and 3 hours with a consistent decrease in the maximal number of binding sites (Bmax) in the submaxillary gland. Furthermore, a marked and prolonged decrease in pilocarpine induced salivary secretion in rats was observed by oral oxybutynin. In contrast, intravesical instillation of oxybutynin produced a significant increase in Kd for specific [3H]NMS binding in the bladder of rats at 0.5 to 4 hours later and also in the submaxillary gland only at 0.5 hours later. The enhancement in Kd was much larger and longer lasting in the bladder than in the submaxillary gland. Moreover, intravesical oxybutynin had little muscarinic receptor binding activity in the heart and colon, and little significant suppression of pilocarpine induced salivation in rats. The plasma concentrations of oxybutynin and N-desethyl-oxybutynin were much higher in rats receiving oxybutynin orally than intravesically. CONCLUSIONS: Intravesical oxybutynin in rats may cause selective binding of bladder muscarinic receptors via a direct local effect, while oral oxybutynin may exert predominant binding of salivary gland receptors. PMID- 15540790 TI - Efficacy of a vaginal sling procedure in a rat model of stress urinary incontinence. AB - PURPOSE: We validated the efficacy of the vaginal sling procedure for the restoration of leak point pressure (LPP) in the rat model of stress urinary incontinence (SUI). MATERIALS AND METHODS: SUI was created in 10 female Sprague Dawley rats by bilateral pudendal nerve transection (PNT) under urethane anesthesia. Vaginal dissection was performed, followed by placement of a 2 x 0.3 cm strip of Prolene (Ethicon, Somerville, New Jersey) mesh at the mid urethral level. LPP was measured before and after PNT through a suprapubic tube using a Crede maneuver. It was also measured after vaginal dissection (sham sling) and after true sling placement. All procedures and measurements were done at the same experimental setting. In each animal LPP was measured 4 or 5 times and the mean was taken. Pairwise differences in LPP between the true and sham slings were calculated using the Wilcoxon signed rank test with p <0.05 considered significantly different. RESULTS: Mean LPP at baseline in all rats was 65.1 +/- 6.0 cm water. LPP decreased to 42.7 +/- 3.7 cm water after PNT. The sham sling further decreased LPP to 26.5 +/- 2.2 cm water, whereas the true sling restored LPP to 56 +/- 4.8 cm water. LPP after true sling placement was not different from baseline (p = 0.16), whereas LPP after sham sling placement was significantly different from baseline (p = 0.002). CONCLUSIONS: This rat vaginal sling model represents a valid surgical method for LPP restoration in the rat model of SUI. This animal model could be used for future studies related to SUI and its treatment relevant to sling surgery. PMID- 15540791 TI - Block of external urethral sphincter contraction by high frequency electrical stimulation of pudendal nerve. AB - PURPOSE: High frequency electrical stimulation (1 to 10 kHz) of the pudendal nerve was evaluated as a method to block the external urethral sphincter contractions and increases in intraurethral pressure induced by electrical stimulation of pudendal nerve efferent axons. MATERIALS AND METHODS: Two stainless steel electrodes were positioned 5 to 10 mm apart on the decentralized pudendal nerve in alpha-chloralose anesthetized cats. The most central electrode was electrically stimulated (40 Hz) to activate the efferent input to the external urethral sphincter. The change in the neurally evoked urethral responses by high frequency electrical stimulation of the distal electrode using a sinusoidal waveform or biphasic charge balanced pulses was examined. RESULTS: For the 2 waveforms stimulation frequencies between 6 and 10 kHz were most effective in blocking external urethral sphincter activation. Intraurethral pressures caused by contractions of the external urethral sphincter were decreased more than 90% by high frequency stimulation using effective frequencies. When high frequency stimulation was turned off, external urethral sphincter responses recovered rapidly. CONCLUSIONS: Reversible block of the external urethral sphincter contractions by high frequency electrical stimulation of the pudendal nerves is a potential method for suppressing detrusor-sphincter dyssynergia and improving voiding in spinal cord injured patients. PMID- 15540792 TI - Localization of aquaporin-7 in human testis and ejaculated sperm: possible involvement in maintenance of sperm quality. AB - PURPOSE: We have previously reported the expression of aquaporin-7 (AQP7) in the rat testis and cloned human AQP7 from the testicular cDNA library. However, to our knowledge the spatial expression pattern and biological roles of AQP7 remain to be elucidated in humans. MATERIALS AND METHODS: We investigated AQP7 expression in the human testis and ejaculated sperm from fertile men and from infertile patients. RESULTS: AQP7 was expressed at the tail of spermatids and spermatozoa in the human testis. AQP7 protein was also detected at the middle piece and the anterior tale portion of ejaculated sperm. However, some infertile patients lacked AQP7 expression in ejaculated sperm, although all fertile men expressed AQP7 protein. The motility rate of AQP7 negative sperm was significantly lower than that of AQP7 positive sperm, while the sperm concentration was not different between AQP7 positive and negative subjects. CONCLUSIONS: AQP7 shows a spatial expression pattern in the human testis. AQP7 may be involved in the maintenance of sperm motility. Furthermore, a lack of AQP7 expression in sperm may be an underlying mechanism of male infertility. PMID- 15540793 TI - Pig kidney: anatomical relationships between the intrarenal arteries and the kidney collecting system. Applied study for urological research and surgical training. AB - PURPOSE: We present a systematic study of the anatomical relationships between intrarenal arteries and the kidney collecting system in pigs. MATERIALS AND METHODS: The intrarenal anatomy (collecting system and arteries) was studied in 91, 3-dimensional endocasts of the kidney collecting system together with the intrarenal arteries. RESULTS: Some anatomical details that have importance to help research and surgical training in urology when using the pig as an animal model were observed and described. It was found that there was only 1 artery per kidney. This artery divided into cranial and caudal branches in 85 cases (93.4%). In 6 cases (6.6%) the primary division of the renal artery was in a dorsal and in a ventral branch. In all cases 2 arteries (1 ventral and 1 dorsal) involved the cranial caliceal group. In the dorsal mid zone a dorsal artery originated from the cranial division of the renal artery, which was obliquely positioned in 47.25% of cases. The arterial supply related to the ventral mid zone of the kidney consisted of branches that coursed horizontally in the ventral surface of the renal pelvis in 81.32% of cases. The caudal division of the renal artery supplied the ventral and dorsal surfaces of the caudal caliceal group in 84.62% of cases, while in 15.38% a dorsal artery supplied its dorsal surface. CONCLUSIONS: Although the results of renal and intrarenal anatomy in pigs could not be completely transposed to humans, many similarities in the pig and human intrarenal arteries support its use as the best animal model for urological procedures. PMID- 15540794 TI - Re: Cost-effectiveness of zoledronic acid for the prevention of skeletal complications in patients with prostate cancer. PMID- 15540795 TI - Re: Intravesical nitric oxide production discriminates between classic and nonulcer interstitial cystitis. PMID- 15540796 TI - Re: Extracorporeal shock wave therapy for Peyronie's disease: exploratory meta analysis of clinical trials. PMID- 15540797 TI - Re: Does bladder preservation (as a surgical principle) lead to retaining bladder function in bladder/prostate rhabdomyosarcoma? Results from Intergroup Rhabdomyosarcoma Study IV. PMID- 15540798 TI - Alcohol, oxidative stress, and free radical damage. AB - Reactive oxygen species (ROS) are small, highly reactive, oxygen-containing molecules that are naturally generated in small amounts during the body's metabolic reactions and can react with and damage complex cellular molecules such as fats, proteins, or DNA. Alcohol promotes the generation of ROS and/or interferes with the body's normal defense mechanisms against these compounds through numerous processes, particularly in the liver. For example, alcohol breakdown in the liver results in the formation of molecules whose further metabolism in the cell leads to ROS production. Alcohol also stimulates the activity of enzymes called cytochrome P450s, which contribute to ROS production. Further, alcohol can alter the levels of certain metals in the body, thereby facilitating ROS production. Finally, alcohol reduces the levels of agents that can eliminate ROS (i.e., antioxidants). The resulting state of the cell, known as oxidative stress, can lead to cell injury. ROS production and oxidative stress in liver cells play a central role in the development of alcoholic liver disease. PMID- 15540799 TI - Dangerous byproducts of alcohol breakdown--focus on adducts. AB - Alcohol breakdown in the liver results in the generation of the reactive molecule acetaldehyde and, as a byproduct, highly reactive oxygen-containing molecules known as oxygen radicals. Both acetaldehyde and oxygen radicals can interact with proteins and other complex molecules in the cell, forming hybrid compounds called adducts. Other adducts are formed with aldehyde molecules, which are produced through the interaction of oxygen radicals with lipids in the cells. Adduct formation impedes the function of the original proteins participating in the reaction. Moreover, the adducts may induce harmful immune responses. Both of these effects may account for some of the damage observed in alcoholic liver disease. Adduct formation has been shown to occur in the livers of humans and animals consuming alcohol and to start and predominate in those liver regions that show the first signs of liver damage. PMID- 15540800 TI - Energy availability and alcohol-related liver pathology. AB - Alcohol consumption alters the metabolism of the most common type of cell found in the liver, the hepatocyte. The presence of alcohol in the body causes the liver to use more oxygen-for example, when breaking down the alcohol. Increased oxygen use, in turn, causes oxygen deficits in several key cells, particularly in hepatocytes located near the small hepatic veins. These veins return blood to the heart for re-oxygenation after it has passed through the liver. Hepatocytes surrounding these veins are the first to show signs of liver disease. The damage induced by oxygen deficits may be exacerbated by alcohol-induced deficits in other components that are essential for cell survival. For example, adenosine triphosphate (ATP), the cell's main source of energy, is generated primarily during the course of two sets of metabolic reactions: glycolysis and the mitochondrial oxidative phosphorylation process. Alcohol consumption may interfere with both of these pathways of ATP production through several mechanisms. An inadequate supply of ATP impairs the cell's ability to perform critical functions, including the repair of alcohol-induced cell damage, and may therefore contribute to cell death and alcoholic liver disease. PMID- 15540801 TI - Endotoxin and Kupffer cell activation in alcoholic liver disease. AB - One central component in the complex network of processes leading to the development of alcoholic liver disease is the activation of immune cells residing in the liver (i.e., Kupffer cells) by a substance called endotoxin, which is released by bacteria living in the intestine. Alcohol consumption can lead to increased endotoxin levels in the blood and liver. When activated, Kupffer cells produce signaling molecules (i.e., cytokines) that promote inflammatory reactions as well as molecules called reactive oxygen species (ROS), which can damage liver cells. Endotoxin activates Kupffer cells by interacting with a complex of protein molecules that are located on the outside of the Kupffer cell or which extend into the cell. Binding of endotoxin alters the activities of the proteins in this complex so that they trigger a cascade of biochemical signals in the Kupffer cell, resulting in cytokine and ROS production and, ultimately, liver damage. Because alcohol can enhance endotoxin release and, therefore, Kupffer cell activation, novel approaches to inhibit these processes might help prevent or ameliorate alcoholic liver disease. PMID- 15540802 TI - Cytokines--central factors in alcoholic liver disease. AB - Many processes related to the consumption or breakdown of alcohol that contribute to alcohol-induced liver disease are mediated by small proteins known as cytokines, which are produced and secreted by liver cells and many other cells throughout the body. Through a variety of actions, cytokines regulate certain biochemical. processes in the cells that produce them as well as in neighboring cells. For example, in case of an infection, they attract white blood cells to the tissues, triggering an inflammatory response. In the liver, persistent cytokine secretion resulting in chronic inflammation leads to conditions such as hepatitis, fibrosis, and cirrhosis. Cytokines also regulate a process known as programmed cell death, or apoptosis, which is in part responsible for alcohol induced destruction of liver tissue. Two cytokines-tumor necrosis factor alpha and transforming growth factor beta-play prominent roles in apoptosis. Finally, a cytokine network mediates the harmful effects of a bacterial protein called endotoxin on the liver. Because of their diverse functions, cytokines might make attractive targets in the prevention or treatment of alcoholic liver disease, and researchers already have obtained encouraging results when testing such approaches. PMID- 15540804 TI - Animal models of alcoholic liver disease--focus on the intragastric feeding model. AB - The use of animal models has contributed to greater understanding of how alcoholic liver disease (ALD) develops, and of how the severity of liver injury is influenced by factors other than alcohol, such as nutrition, oxygen deprivation (as occurs with sleep apnea or smoking), and gene regulation. This article focuses on the use of one animal model in particular, the intragastric feeding model in rats. This model allows scientists to rigorously control an animal's consumption of both alcohol and dietary nutrients and is providing important information on the mechanisms of injury of alcoholic liver disease. PMID- 15540803 TI - Intracellular proteolytic systems in alcohol-induced tissue injury. AB - The body constantly produces proteins and degrades proteins that are no longer needed or are defective. The process of protein breakdown, called proteolysis, is essential to cell survival. Numerous proteolytic systems exist in mammalian cells, the most important of which are the lysosomes, the ubiquitin-proteasome pathway, and enzymes called calpains. Lysosomes are small cell components that contain specific enzymes (i.e., proteases) which break down proteins. Alcohol interferes with the formation and activity of lysosomes and thus may contribute to protein accumulation in the liver, which can have harmful effects on that organ. In the ubiquitin-proteasome pathway, proteins that are to be degraded are first marked by the addition of ubiquitin molecules and then broken down by large protein complexes called proteasomes. Alcohol impairs this proteolytic system through several mechanisms, possibly leading to inflammation and even cell death. Calpains are proteases that are involved in several physiological processes, including the breakdown of proteins that give cells their shape and stability. In contrast to the lysosomal and ubiquitin-proteasome systems, calpains in brain cells are activated by alcohol, to potentially detrimental effect. PMID- 15540805 TI - J-aggregates on metal nanoparticles characterized through ultrafast spectroscopy and near-field optics. AB - Illumination of metal nanoparticles at the plasmon resonance produces enhanced evanescent fields on the nanoparticles' surfaces. The unusual strength of the field makes it a target for exploring photoinduced phenomena at the nanoscale, if efficient functionalization or coating of the nanoparticle surface with appropriate chromophores is possible. One direction is to use cyanine dyes that form monolayers of J-aggregates on the surface of noble metal nanoparticle colloids. The unique, collective electronic properties of J-aggregates produce excitons with enormous extinction coefficients that are of interest for their efficient energy transfer, electron transfer, and nonlinear optical properties. In that vein, we report our results on time-resolved spectroscopy and near-field scanning optical microscopy (NSOM) of J-aggregate exciton dynamics on Ag and Au nanoparticle colloids. Ultrafast transient absorption studies show that J aggregate exciton lifetimes on Ag nanoparticles are much longer than on Au nanoparticles, with a 300 ps lifetime that is two orders of magnitude longer than the electronic processes in the nanoparticles themselves. Complementary NSOM studies of the colloids show that fluorescence from the J-aggregates on the Ag nanoparticles is induced by the scanning probe. These results may be significant for improving the nanophotonic performance of hybrid materials for nanoscale applications. PMID- 15540806 TI - Near-field spectroscopy of water-soluble and water-insoluble porphyrin J aggregates. AB - Mesoscopic properties of J-aggregates of water-soluble [tetrakis(4 sulfonatophenyl)porphyrin] and water-insoluble [tetrakis(4 methoxyphenyl)porphyrin and tetraphenylporphyrin] tetraphenylporphyrin derivatives in thin films have been investigated by scanning near-field optical microscopy. From surface topography, the sample films were found to be composed of planar J-aggregate microcrystallines. The transmittance spectra of the water insoluble samples are site specific, while those of the water-soluble samples are not. This observation might reflect the spatial inhomogeneity of the thin films in mesoscopic scales and is related to the broad J-bands of water-insoluble samples. Excited state lifetimes, obtained from time-resolved, pump-probe measurements in small domains, were in the 30-70 ps range for 4-methoxy substituted and in the 40-100 ps range for unsubstituted tetraphenylporphyrin, which are of the same order as the reported far-field results for 4-sulfonated compound in water. PMID- 15540807 TI - Localization of surface plasmon by using a metal-coated axicon prism. AB - We present the localization of surface plasmon using a metal-coated axicon prism and its application to fluorescence scanning microscopy. The cone angle of the axicon prism is designed such that a plane wave incident on the prism can excite surface plasmon on the side surface of the prism. Since the surface plasmon propagates toward the apex of the prism, an extremely high enhancement of the electric field is expected. The proposed technique can overcome the trade offbetween the field-enhancement factor of surface plasmon and the resolution limit due to the propagation of surface plasmon. The experimental result obtained with the localized surface plasmon is also demonstrated. PMID- 15540808 TI - Discrete polarization absorption property of carbon nanotubes studied by using scanning near-field optical microscope. AB - By using scanning near-field optical microscopy (SNOM), we have observed the polarization phenomena of carbon nanotubes (CNTs), which were synthesized by pyrolysis of a metal phthalocyanine method. Using SNOM, an area of about 100 nm in the thin part of the honeycomblike aligned CNT can be investigated, where the CNTs are nearly parallel or cross aligned with only a few layers. Transmission intensity of the light absorption with constant height scan identified the thickness of the nanotube layers. By changing the linear polarization of the incident light, the dependence of transmission versus polarization angle was recorded. The observed different polarization cannot be explained by using the model of optical absorption of continuum medium. We have proposed a model of discrete polarization absorption, which can be used to identify the number of layers of CNTs and their relative orientations. The combination of SNOM technique and our model can be used for optical polarization of carbon nanotubes of discrete medium. PMID- 15540809 TI - Near-field optical technique applied for investigation of the characteristics of polymer fiber and waveguide structures. AB - This article summarizes the near-field optical technique applied for investigating the characteristics of polymer fiber and waveguide structures. The near-field optical technique is used to analyze multimode interference structures of fiber. The localized fluctuation of the transmission caused by fractal cluster is carried out in Nd3+- and Eu3+-doped polymer fiber and film by means of a scanning near-field optical microscopy. The near-field optical spectrum of Nd3+ doped polymer fiber is investigated. The topography and near-field intensity images of Azo-polymer liquid crystal film for waveguide are obtained simultaneously. PMID- 15540810 TI - In-situ and high-resolution optical observation in local anodic oxidation process using a scanning near-field optical microscope. AB - Using a local oxidation process with a probe tip of a scanning near-field optical microscope (SNOM), antimony oxide structures with a pattern width of several 100 nm were fabricated on antimony thin films. We also investigated the characteristics of the oxidation process using the transmission mode SNOM. Although slight topographic changes due to oxidation were observed by atomic force microscopy, an increase in transmission due to the phase change of the metal to transparent oxide was observed. The depth of the oxide embedded in the antimony film was estimated from the change of the transmission intensity without destroying the sample. This fabrication method is appropriate for making a prototype of future nano-optical devices and for trimming them in order to optimize their functions on demand. PMID- 15540811 TI - Topography-induced polarization anisotropy in mesoscale structures. AB - Optical polarization anisotropy on the surface of subwavelength air-dielectric structures was studied by polarization-modulation near-field optical microscopy. A large degree of polarization anisotropy was measured in the higher topographic region of transparent gratings. In two-dimensional air-hole arrays, the largerpolarization anisotropy occurs in the dielectric regions. Both dielectric and air regions have a 90 degrees difference in the directions of maximum transmission. From calculations with the finite-difference time-domain method, we verified that diffracted light from the mesoscale topographic edges contributes to polarization anisotropy and directional differences. PMID- 15540812 TI - Imaging near-field transverse modes of vertical-cavity surface-emitting lasers by near-field scanning optical microscopy. AB - We have characterized the oxide-confined vertical-cavity surface-emitting laser (VCSEL) using short-tip, tapping-mode tuning fork near-field scanning optical microscopy (TMTF-NSOM). The near-field radiation patterns of the VCSEL were measured. By comparing the topographic and optical images, we attribute the asymmetric transverse modes to the geometric defect outside the oxide aperture. We also performed spatially resolved spectroscopic imaging over the surface of the VCSEL by coupling NSOM to a spectrometer. PMID- 15540813 TI - Near-field imaging of the interactions of evanescent fields. AB - The near-field images of interferences generated by the reflection or by the interactions of four evanescent fields of two different wavelengths are obtained. The near-field interactions of the evanescent fields are studied. PMID- 15540814 TI - Probing the focused laser spot of the apodized light source. AB - This paper deals with both theoretical and experimental studies of near-field imaging of the focused laser spot. In theoretical simulation, we calculate the intensity distribution in the focal region of the apodized light source. Size reduction of the focusing spot of an annular light beam is found to be related with various epsilon, which is the ratio of the obstruction radius to the radius of lens aperture. The experimental setup is based on a near-field scanning optical microscope (NSOM) system. An opaque disk is used to make an annular beam. A high numerical aperture objective lens (NA = 0.8) is used to focus incident lights of different annular light beams, and a tapered near-field optical fiber probe of the NSOM system is used to collect the intensity of the focal field. Results demonstrate that a better transverse resolution can be obtained from the reduction of the focusing spot. The intensity profile of an annular beam may have a significant influence on the size reduction of the focusing spot. PMID- 15540815 TI - Numerical and experimental research on the near-field optical virtual probe. AB - A near-field optical virtual probe (NFOVP) is a type of immaterial tip based on the principle of near-field evanescent wave interference. Evanescent wave interference and the aperture type play very significant roles in generating near field optical virtual probes. Two evanescent waves, propagating in opposite directions, will interfere to generate the confinement field. The central peak of the wave distribution carries the preponderance of energy. An aperture can be used to suppress the sidelobe in the energy distribution while forming the NFOVP. In this paper, the NFOVP is investigated numerically by means of the three dimensional (3-D) finite-difference time-domain (FDTD) method. Some significant factors in the creation of the NFOVP, such as the shape and size of the aperture, the incident light, and the specimen (nanoparticle), are studied and discussed. The optical field of the evanescent wave interference has been measured by near field scanning optical microscopy (NSOM), and the preliminary experimental results are shown. PMID- 15540816 TI - Apertureless near-field scanning optical microscope working with or without laser source. AB - An apertureless near-field scanning optical microscope (ANSOM), used indifferent configurations, is presented. Our versatile home-made setup, based on a sharp tungsten tip glued onto a quartz tuning fork and working in tapping mode, allows to perform imaging over a broad spectral range. We have recorded optical images in the visible (wavelength, lambda = 655 nm) and in the infrared (lambda = 10.6 microm), proving that the setup routinely achieves an optical resolution of <50 nm regardless of the illumination wavelength. We have also shown optical images recorded in the visible (lambda = 655 nm) in an inverted configuration where the tip does not perturb the focused spot of the illumination laser. Approach curves as well as image profiles have revealed that on demodulating the optical signal at higher harmonics, we can obtain an effective probe sharpening which results in an improvement of the resolution. Finally, we have presented optical images recorded in the infrared without any illumination, that is, the usual laser source is replaced by a simple heating of the sample. This has shown that the ANSOM can be used as a near-field thermal optical microscope (NTOM) to probe the near field generated by the thermal emission of the sample. PMID- 15540817 TI - Design and experiments of a near-field optical disk head for very high efficiency. AB - A high-density optical disk system with super parallel optical heads using a vertical cavity surface emitting laser (VCSEL) array is described for higher data transfer rate and technological capability. Optical heads of the VCSEL array and microlens array play key roles for obtaining higher evanescent light from small apertures with the optical disk, which is coated with lubricant and protective films on the flat surface in order to keep the gap between the super parallel optical head and the disk surface within 20 nm. Higher throughput efficiency has been obtained in the near-field semiconductor optical probe array head. However, the obtained evanescent light power is about 10 microW from a 100 nm probe aperture and 1 m W VCSEL power, which is still not enough to write a bit on the phase change optical disk. One solution to improve the optical power more than 10 times is to develop a special nanofabricated optical probe array of higher throughput efficiency. A metal fine grating fabrication method to get evanescent light wave resonant enhancement has been studied along with a 2-dimensional finite difference time domain simulation technique. PMID- 15540818 TI - Simulation and fabrication of subwavelength structures for a nanometer feature enabling lens-less laser writers. AB - A set-up was proposed to propagate submicrometer light beams without resorting to the conventional approach of using a lens. The main target was to develop a method of either by passing or circumventing the diffraction limit to develop a new optical head for a laser writer system with submicrometer resolution. Starting from using surface plasma theory to explain the mechanism of extraordinary transmission phenomenon, we move to discuss the fabrication processes for free-standing metal films and the corresponding transmission light beam images in the far field. A finite difference time domain (FDTD) method was adopted to simulate the electromagnetic field distribution of transmission light beams from the near field to the far field. The experimental results confirm that surface structures on the incident side of incoming light beams can excite surface plasma and enhance the throughput energy of transmission light beams. In addition, we also established that surface structures on the exit side can suppress the diffraction effect and make transmission light beams directional. The experimental and simulation results were found to agree well with each other, which validates the effectiveness of the FDTD simulation approach. As a free standing film was too fragile to be of practical use, a proposition to develop a substrate-based nanostructured film to ease the construction of an optical head for a laser writer was developed. PMID- 15540819 TI - Second harmonic generation of biotin and biotin ester microcrystals trapped in optical tweezers with a mode-locked Ti:sapphire laser. AB - Intense second harmonic generation (SHG) was observed from microcrystals of biotin and biotin ester trapped by optical tweezers, formed with a focused near infrared pulsed laser beam. The intensity of SHG depends strongly on the states of the microcrystals and the excitation wavelength. Microscopic scanning images of biotin and biotin ester were obtained in high contrast with SHG. Simultaneous trapping and excitation of SHG and two-photon autofluorescence of biotin and biotin ester microcrystals allow us to investigate their structure and optical properties. These optically trapped particles (of submicron size) are useful as nonintrusive microscopic probes for high-resolution studies. PMID- 15540820 TI - Fabrication and optical studies of monolayer assemblies of Au nanoparticles. AB - Two-dimensional nanoparticle self-assemblies provide a means of tuning the optical properties by manipulation of the fabrication parameters, thereby controlling the light-matter interaction. We report fabrication of self-assembled monolayers of colloidal gold nanoparticles on organosilane-coated substrates and a study of the linear and nonlinear optical properties. With increase in particle coverage of the gold monolayer self assemblies, an additional band in the absorption spectrum due to the collective particle surface plasmon resonance could be seen. The intensity and lambda(max) of this feature depends on the interparticle spacing and particle aggregation. Z-scan measurements show a saturable absorption behavior and a negative nonlinear refractive index of the monolayer films. A strong correlation between particle density and the linear and nonlinear optical properties is established. PMID- 15540821 TI - Study of optical transmittance of AgOx nano thin film in super-resolution near field structure. AB - The AgOx-type super-resolution near-field structure (super-RENS) is one of the nano thin films that can give a nonlinear optical effect similar to that of the optical probe of the scanning near-field optical microscope. Structural variations of different thicknesses of the AgOx thin film are observed and studied using a field-emission scanning electron microscope (FE-SEM). For single AgOx nano thin film, the critical temperature and threshold of the evaporation temperature are 154 degrees and 600 degrees, respectively. Results show that the sandwiched AgOx nano thin film has a higher critical temperature, that is, about 280 degrees. For the sandwiched AgOx nano thin film, the transmittance peak is dependent on the thickness of the AgOx thin film. The interesting properties of the AgOx nano thin film of the super-resolution structure indicate a great potential for photonic applications. PMID- 15540822 TI - Evanescent signal detection and enhanced resolution with random silver nanoparticles. AB - Inspired by the subwavelength resolution capability of the super-resolution near field structure, a simplified optical structure was proposed to investigate the complicated near-field optical interactions between random silver nanoparticles and evanescent waves from a zeroth-order grating. The simplified structure consisted of a glass thin film embedded with random silver nanoparticles on a zeroth-order glass grating. The near-field and far-field optical properties of this structure were investigated by the finite-difference time-domain (FDTD) numerical simulations and the optical resolution beyond diffraction limit was demonstrated. PMID- 15540823 TI - Enhanced resolution of AgOx-type super-RENS disks with periodic silver nanoclusters. AB - The super-resolution near-field structure (super-RENS) of the AgOx-type can perform the task of the high-density near-field optical recording in a more feasible way and has demonstrated better signal-to-noise ratio. For further exploration of the optical resolution and controllability of the super-RENS disks, we studied the near-field and far-field optical properties of the AgOx type super-RENS embedded with periodic silver nanoparticles or nanoclusters with finite-difference time-domain (FDTD) simulations. It was found that the period of nanoclusters could selectively manipulate the resolution of the super-RENS disk. PMID- 15540824 TI - Near- and far-field optical properties of embedded scatters in AgOx-type super resolution near-field structures. AB - In recent near-field optical recording techniques, the super-resolution near field structures (super-RENS) have been successfully demonstrated to overcome the diffraction limit. To realize the possibility of replacing the conventional near field optical probe by the super-RENS, and to understand the relations between the near-field enhancements and detectable far-field signals, we use the two dimensional finite-difference time-domain (FDTD) method to study the near-field and far-field properties of different types of embedded scatters in the AgOx-type super-RENS. PMID- 15540825 TI - Dispersion relations of plasmon wave in metallic nanowires. AB - The dispersion relations of 0th transverse-magnetic (TM) mode plasmon wave propagating along silver nanowires are calculated. Based on these relations, the propagation loss, phase velocity, and group velocity of the plasmon wave are extracted to study the effects of nanowire diameter and surrounding dielectric medium. Several useful inferences for the design of an optical waveguide in nanometer scales are drawn from these results. The dispersion relations of high order hybrid modes of metallic nanowires are being investigated at the present stage. This study will be useful for the design of nanometer-scaled metallic waveguides. PMID- 15540826 TI - Numerically investigating near-field scattering for spatial enhancement of single irregular nano plasma particle. AB - Near-field scattering from a nano silver particle has been studied with an optical wave passing through it. The scattered field was found to be enhanced around the semisphere on which the incident wave impinges. Comparing it with the spherical particle, the irregular shape of a nanoparticle only changes the distribution of the scattered field. In this investigation, approached by finite difference time domain (FDTD), it was revealed that the peak value of the scattered field is proportional to the particle radius; this might be due to the fact that more plasmons could be excited. Furthermore, the highest enhancement occurred at the incident frequency close to plasma frequency of silver (approximately 2000 THz). Thus, the numerical results could give a guide for optimizing AgOx type super resolution near-field structure (RENS) applications. When the red light source can be replaced with shorter wavelength ones, or larger silver particles can be segregated, the near-field scattered enhancement might result. PMID- 15540827 TI - Localization of optical fields using a bow-tie nanoantenna. AB - Localization of optical fields in a bow-tie nanoantenna is studied using a high frequency structure simulator (HFSS). Simulation results show a strong localization effect in the gap of the antenna. The enhanced intensity in the gap is due to the surface plasmon resonance and the concentration of energy flow. PMID- 15540828 TI - Surface plasmon polariton in visible frequency of a nano-slab consisting of left handed material. AB - We have investigated the behavior of surface plasmon polariton of a left-handed material (LHM) nano-slab. The LHM slab is chosen to operate in visible region, that is, it has negative permittivity and permeability within a frequency range of visible light. The dispersion curves of the LHM slab and the corresponding attenuated total reflection (ATR) spectra have been calculated in this article. PMID- 15540830 TI - Why us? PMID- 15540833 TI - Healthcare ergonomics--what does systemic incident / accident investigation have to do with ergonomics? PMID- 15540832 TI - What CNAs taught us: lessons learned while teaching CNAs how to handle resident aggression. AB - The authors conducted a series of two hour training programs in twenty five nursing facilities involving 479 CNAs. The focus was on how to deal with resident aggression. In the process of presenting 42 programs, we were impressed by the CNAs. In fact, the CNAs taught us about themselves, their work and their lives. This paper shares what the CNAs taught us. PMID- 15540834 TI - A nursing guide to the assessment of GERD in long term care. PMID- 15540835 TI - Anticholinergic effects in older adults. Weighing the risk-benefit ratio. PMID- 15540836 TI - Osteoporosis in long term care. Part 2 of 2: treatment strategies. PMID- 15540837 TI - Bathing safety... Safeguarding the resident and your staff! PMID- 15540838 TI - Quality standards for choosing an adult incontinence product. AB - Urinary incontinence (UI) is the leading cause for institutionalization of the elderly. According to the Agency for Health Care Policy and Research (AHCPR), over 50% of the elderly resident population living in nursing homes are considered to be incontinent. In long term care facilities, 33% of medical supplies purchased are incontinence products. This 33% represents the number one item for this classification of expenses. PMID- 15540839 TI - Integrative care--spiritual care: not just the domain of clergy. PMID- 15540841 TI - Warning! Clinical placement crisis looming! PMID- 15540842 TI - Enrolled nurses treated unfairly. PMID- 15540843 TI - Made where? PMID- 15540844 TI - Work it. Claim it. PMID- 15540845 TI - What nursing shortage? PMID- 15540846 TI - Specialist hepatology nurses leading the way. AB - Hepatitis C virus (HCV) is transmitted when infected blood from one person gets directly into the bloodstream of another. The most common way that HCV is transmitted in Australia is through the sharing of injecting drug use equipment. It is extremely likely that all nurses will meet a person with chronic HCV at some stage of their career. PMID- 15540847 TI - A fine line. PMID- 15540848 TI - Ilizarov correction of malrotated femoral shaft fracture initially treated with an intramedullary nail: a case report. AB - Ideally, acquired femoral malrotation as a complication of intramedullary nailing of a femoral shaft fracture should be identified and corrected early in the course of treatment. In this report, we present a previously undescribed precise surgical technique for acutely correcting an acquired femoral malrotation deformity 20 days after fixation of a femoral shaft fracture with a statically locked intramedullary nail. An llizarov external fixator was used intraoperatively to correct a 27 degree femoral malrotation deformity in a 19 year-old man. PMID- 15540849 TI - Knee arthrodesis after total knee arthroplasty. PMID- 15540850 TI - Total elbow arthroplasty: current indications, factors affecting outcomes, and follow-up results. AB - Indications for total elbow arthroplasty (TEA) are expanding in light of newly published follow-up studies. The unique characteristics of the pathologic condition, appropriate patient selection, suitable choice of prosthesis, and proper surgical technique are critical factors that have a large impact on the ultimate results of TEA for any indication. This article addresses these factors as they relate to various indications and presents a review of the recently published results of the 10-year follow-up studies for TEA. PMID- 15540851 TI - Debridement and primary closure of nonhealing foot wounds. AB - Fifty-eight chronic nonhealing foot wounds (51 patients) were treated with irrigation, aggressive debridement, and primary tension-free closure. Factors such as wound location, wound size, presence of infection, and healing outcome were recorded. In addition, medical comorbidities and preoperative laboratory test results were reviewed. Thirty-seven (64%) of the 58 wounds healed after primary closure. Of the other 21 wounds, 16 healed after repeat irrigation, debridement, and closure or local wound care; 2 patients were lost to follow-up after initial failed wound healing, 1 patient died after initial failed wound healing, and 2 cases were salvaged with amputation. Failed primary closures were thought not to increase wound size; all but 3 of these closures decreased wound size significantly. Differences between the wounds that healed primarily and the wounds that failed healing were not statistically significant. Diabetes was present in 46% of the patients whose wounds healed primarily versus 71% of the patients whose wounds failed healing (P = .06). Irrigation, debridement, and primary closure of nonhealing foot wounds can be a useful treatment option for most such patients. Complete healing or reduced wound size occurs in 95% of cases. PMID- 15540852 TI - Comparison of efficacy of oral rofecoxib and ketorolac in controlling early postoperative outpatient orthopedic surgical pain. AB - We compared the efficacy of rofecoxib and ketorolac in controlling postoperative pain after outpatient surgery. Patients were randomly assigned to receive either rofecoxib or ketorolac preoperatively and every morning for 5 postoperative days. Patients in both groups received oxycodone as needed for breakthrough pain. After 5 postoperative days, patients were asked for information-pain score; number of supplemental analgesics used; and severity of any incision-site bleeding, nausea, or diarrhea. Rofecoxib and ketorolac did not differ on these measures. Rofecoxib and ketorolac are equally effective in controlling postoperative pain. PMID- 15540853 TI - Lipofibromatous hamartoma of the median nerve: case presentation of MRI, ultrasound, electrodiagnostic, histologic, and surgical findings. PMID- 15540854 TI - Isolated compartment syndrome of the extensor carpi ulnaris: a case report. PMID- 15540855 TI - Fracture of the lateral process of the talus: a report of 2 cases. AB - Fracture of the lateral process of the talus is historically a rare injury with an increasing incidence. This fracture is easily misdiagnosed as an ankle soft tissue sprain, and a high index of suspicion is essential in making a timely diagnosis to avoid any long-term sequelae. A CT scan should be performed in preference over an MRI scan. PMID- 15540857 TI - President's message. PMID- 15540858 TI - Endovascular stent insertion for abdominal aortic aneurysm: a nursing perspective. PMID- 15540856 TI - Evaluation and management of acromioclavicular joint injuries. AB - The acromioclavicular joint is stabilized by the coracoclavicular and acromioclavicular ligaments and by the trapezius and deltoid muscles. Joint dislocation commonly results from a direct blow to the acromion. Injury types I through III are generally treated nonoperatively, whereas types IV through VI are treated operatively. Nonoperative protocols should always begin with ice and immobilization. Operative techniques include acromioclavicular ligament repair, dynamic transfer of the conjoined tendon, coracoclavicular ligament reconstruction, and coracoacromial ligament transfer. The goal with any injury type should always be full return to the patient's preinjury condition. PMID- 15540859 TI - Keeping spirit alive at work: taking care of the caregiver. PMID- 15540860 TI - Latex allergy: still a challenge? PMID- 15540861 TI - Life-long dream fulfilled. AB - In the early 1940's, when I was a 14 year old in grade 9 at Chebucto Road School, in Halifax, NS, a Guidance Counsellor taught my class a compulsory subject titled CAREERS. The credit for this subject criteria required development of a 'CAREER BOOK'. The title page for my career book was Nurse As My Career. I had actually chosen this career long before the age of fourteen. PMID- 15540862 TI - One child with hope for the future. AB - CAMTA, the Canadian Association of Medical Teams Abroad, is a group of medical and lay individuals dedicated to helping underprivileged people suffering from orthopedic problems in Ecuador. The informal group conducted two missions prior to officially incorporating in 2001 and has, since its official founding, conducted 3 more missions in Ecuador. During their surgical missions the CAMTA team also aims to share information and provide teaching to local nurses and physicians. PMID- 15540863 TI - What pay for performance can mean for quality managers. AB - Top management wants to see the numbers before the public does. Learn to determine whose data are the most valid and accurate. Consumers evaluate hospitals far differently than the Joint Commission on Accreditation of Health Care or The Centers for Medicare & Medicaid. PMID- 15540864 TI - Take proactive steps with pay for performance. PMID- 15540865 TI - HealthGrades opening up methodology to review. AB - Workshop topics include risk-adjustment methodology and statistical models. The science behind this type of health care data still is in the nascent stage. Use by employers and consumers should drive quality improvement. PMID- 15540866 TI - Researchers unveil tool to predict cardiac death risk. AB - Calculating tool can fit on a pocket card or be downloaded onto a PDA. The data are current, having been collected through December 2003. The GRACE model is based on data from 15,007 patients who were followed for six months. PMID- 15540867 TI - Evidence-based design could help quality of care. AB - Design can help reduce infection, risk, injuries from falls, and medical errors. Stress levels can be lessened for patients, families, and staff. Hospital building boom under way provides significant opportunities. PMID- 15540869 TI - New surgery standards could save 8,000 lives. PMID- 15540868 TI - Children in hospitals often have adverse events. AB - A new study found a total of 51,615 patient safety events involving children in hospitals during 2000. The study quantifies impact of patient safety events in terms of excess hospital stays and charges. Postoperative respiratory failure increased death rate as much as 76%. PMID- 15540870 TI - Sorting things out. PMID- 15540871 TI - Tears, laughter, and "warm fuzzies". PMID- 15540872 TI - Need for more hospice research. PMID- 15540873 TI - Self-identifying sexuality. PMID- 15540874 TI - Do atypical antipsychotic agents cause tardive dyskinesia? PMID- 15540875 TI - Primary care update for mental health nurses: evidence-based guidelines for nursing assessment, intervention, and follow up. AB - Physical health comorbidities are often undiagnosed or undertreated in individuals with severe and persistent mental illness. Mental health nurses are in an ideal position to identify, refer, and monitor treatment of these comorbidities. National evidence-based practice guidelines provide key information about common physical health comorbidities that mental health nurses can readily use in their everyday practice. PMID- 15540876 TI - Adolescent postabortion groups: risk reduction in a school-based health clinic. AB - A short-term postabortion group for adolescents was developed. Three groups were conducted in an adolescent mental health clinic within an urban high school-based health clinic. The clinical group experiences offered the adolescents an opportunity to integrate the experience of pregnancy and the abortion decision into their lives. At follow up, adolescents who participated in th postabortion counseling group indicated that they chose and used a method of birth control, did not repeat an unplanned pregnancy, and remained in high school. PMID- 15540877 TI - My life on meds. PMID- 15540878 TI - I got this!--preparing NMA for America's needed healthcare revolution/NMA, America: making our nation's healthcare system equitable. PMID- 15540879 TI - Physician race and ethnicity, professional satisfaction, and work-related stress: results from the Physician Worklife Study. AB - There are limited data about minority physicians' professional satisfaction and job stress. In this study, we describe by race and ethnicity, satisfaction, and job stress among a national sample of physician. We analyzed data from 2,217 respondents to the Physicians' Worklife Survey (PWS), a career satisfaction survey of physicians drawn from the AMA Physician Masterfile. Scales measuring overall job and career satisfaction and work-related stress were constructed from Likert-response items. We examined the association between physician ethnicity and each of these scales. Respondents included 57 black, 134 Hispanic, 400 Asian or Pacific Islander, and 1,626 white physicians. In general, minority physicians appeared to serve a more demanding patient base than did white physicians. Hispanic physicians reported significantly higher job (p=0.05) and career (p=0.03) satisfaction compared to white physicians but no significant difference in stress. Asian or Pacific Islander physicians averaged lower job satisfaction (p=001) and higher stress (p<0.01) compared to white physicians. Black physicians did not differ significantly from white physicians on any of the three measures. Significant racial and ethnic variations were found with respect to several specific satisfaction domains: autonomy, patient care issues, relations with staff, relations with the community, pay, and resources. PMID- 15540880 TI - Uncovering myths and transforming realities among low-SES African-American men: implications for reducing prostate cancer disparities. AB - PURPOSE: Prostate cancer provides the most dramatic evidence of cancer disparities based on race and ethnicity among U.S. men. African-American men still hold a commanding lead in both prostate cancer incidence and mortality, particularly among those of low socioeconomic status (SES) and the medically underserved. Therefore, the need for early intervention persists. The purpose of this exploratory pilot study was to: a) assess the knowledge of a cohort of low SES African-American men regarding prostate health/prostate cancer, and b) uncover myths/misinformation as barriers to prostate health decisions and behaviors. PROCEDURES: Asymptomatic African-American men participated in focus groups to candidly discuss: a) health concerns, b) prostate health, c) prostate cancer screening, diagnosis and treatment, and d) factors influencing prostate health decisions/behaviors. FINDINGS: Participants revealed sociocultural and psychological barriers: myths and lack of accurate/adequate knowledge about prostate health and cancer, fear, denial and apathy. CONCLUSIONS: These findings suggest factors that may explain the reluctance and limited participation in prostate health and prostate cancer services among medically underserved, socioeconomically disadvantaged, African-American men. Lack of knowledge, which affects all barriers to care, is amenable to change. Therefore, improvements in prostate cancer outcomes are achievable through culturally and linguistically appropriate health education tailored to their specific needs. PMID- 15540881 TI - Clinical studies of thiazide-induced hyponatremia. AB - To determine the prevalence and vulnerability of symptoms from thiazide-induced hyponatremia, we reviewed 223 cases of symptomatic hyponatremia enrolled between January 1996 and April 2002. There was a high frequency of neurologic manifestation of thiazide-induced hyponatremia, whereas clinical dehydration was not a discernible feature. Female patients presented with lower serum sodium levels than male counterparts (114+/-8 versus 117+/-8 mmol/L, P=0.02), although the frequency of central nervous system manifestation was comparable between two gender groups. The most frequent symptoms were malaise and lethargy (49%), followed by dizzy spells (47%) and vomiting (35%). Degree of hyponatremia upon presentation predicted the development of confusion and vomiting symptoms. Serum sodium concentration < or =115 mmol/L was significantly associated with the development of confusion (odds ratio 2.6, 95% confidence interval 1.3 to 5.1, P=0.004). Our results show that symptoms from thiazide-induced hyponatremia primarily reflect osmotic water shift into brain cells rather than extracellular fluid volume depletion. PMID- 15540882 TI - Quality care improvement program in a community-based participatory research project: example of Project DIRECT. AB - A continuous quality care improvement program (CQIP) was built into Project DIRECT (Diabetes Interventions Reaching and Educating Communities Together) to improve providers' patterns of diabetes care and patients' glycemic control. Project DIRECT consisted of a comprehensive program aimed at reducing the burden of diabetes in the vulnerable high-risk African-American population of southeast Raleigh, NC. Forty-seven providers caring for this target population of adult diabetes patients were included in this quasi-experimental study. At the initial session, providers learned about the CQIP components, completed a planning worksheet, and chose a CQIP coordinator. Educational events included continuing education in practices and through conferences by experts, and guideline distribution. Follow-up was accomplished through phone calls and visits. Effectiveness was measured by a change in prevalence of selected patterns of care abstracted from 1,006 medical charts. Appropriate statistical methods were used to account for the cluster design and repeated measures. At the fourth follow-up year, approximately 40% of providers still participated in the program. Among the providers who stayed in the program for the whole study period, most selected quality care patterns showed significant upward trends. Glycemic control indicators did not change, however, despite an increased number of hemoglobin A1c tests per year. A diabetes CQI program can be effectively implemented in a community setting. Improved performance measures were not associated with improved outcomes. These results suggest that a patient-centered component should reinforce the provider-centered component. PMID- 15540883 TI - Screening for diabetes in an African-American community: the Project DIRECT experience. AB - AIM: To report the results of a community-based screening program associated with Project DIRECT, a multiyear diabetes mellitus prevention and control project targeting African-American residents of southeast Raleigh, NC. METHODS: Between December 1996 and June 1999, 183 screening events took place in community settings. Screening was by capillary glucose concentration. Participants with a positive screen were referred for confirmatory testing and physician follow-up. MAIN RESULTS: Risk factors for diabetes were prevalent, including ethnic minority race (88.2%), obesity (45.6%), and family history of diabetes (41.7%). In all, 197 personshad an elevated screening result; the prevalence of diabetes in the screened population that underwent follow-up testiing was 1.7%. Despite persistent tracking efforts, 28% of the persons with a high screening test received no final diagnosis CONCLUSIONS: In this community-based screening program targeted to high-risk African Americans, risk factors for diabetes were common, but new cases of undiagnosed diabetes among participants were uncommon. Intensive follow-up for persons with high screening values is necessary but difficult to achieve. Our results support national recommendations against community-based screening; opportunistic screening for diabetes in clinical settings is likely a more effective use of resources. PMID- 15540884 TI - The Tripler Army Medical Center's LE3AN program: a six-month retrospective analysis of program effectiveness for African-American and European-American females. AB - This is a retrospective study that examines the effectiveness of the Tripler Army Medical Center (TAMC) LE3AN Program for weight management among African-American and European American women. African-American and European-American active-duty females who enrolled in the TAMC LE3AN Program between July 1998 and December 2001, and completed six months of follow-up were included in the analysis. The results indicate that the program is associated with significant weight loss for participants, and that it is equally effective for African-American and European American women. Weekly follow-up visits were correlated with greater weight loss. PMID- 15540885 TI - The effect of parental race on fetal and infant mortality in twin gestations. AB - Previous work has found that singleton birth outcomes are better if the father is black and the mother is white than if the father is white and the mother is black. We sought to examine the effects of parental race on fetal and infant mortality in twins. We analyzed the fetal and infant mortality rates in four groups [both parents white (W-W), both parents black (B-B), father black and mother white (FB-MW), and father white and mother black (FW-MB)], using the 1995- 1997 U.S. twin registry data (249,221 twins). Compared to W-W, the infant mortality for B-B, FW-MB, and FB-MW (respectively, relative risk [RR] 1.84, 95% confidence interval [CI] 1.73-1.95; RR 1.39, 95% CI 1.03-1.51; and RR 1.49, 95% CI 1.26-1.77) were all significantly different from W-W but not from each other. When fetal mortality was added to infant mortality, the combined mortality was highest for B-B (RR 1.66, 95% CI 1.58-1.75), intermediate for FW-MB (RR 1.18, 95% CI 0.92-1.51) and FB-MW (RR 1.37, 95% CI 1.19-1.58) and lowest for W-W. Thus, twin infants born to black parents have higher risk of fetal and infant mortality compared with twin infants born to white parents and infants of mixed race parents generally have intermediate outcomes. PMID- 15540886 TI - Why I am entering into an executive MBA program. PMID- 15540887 TI - Ethnic and geographic variation in gastrostomy placement among hospitalized older patients. AB - While whites may receive more healthcare services than African Americans, gastrostomies are more commonly placed in older African Americans. The purpose of this study was to explore geographic and ethnic variations in gastrostomy placement among older individuals. Data from National Hospital Discharge Surveys conducted between 1996 and 1999 were analyzed. The overall gastrostomy placement rate (per 1,000 discharges) in individuals aged 65 years or older during the four years was 10.9 (95% Confidence Interval [CI]: 10.3-11.5)--among African Americans 20.0 (95% CI: 17.8-22.2) and among whites 10.2 (95% CI: 9.5-10.9). According to region, placement rates per 1000 discharges were 8.9 (95% CI: 7.6-10.2) in the west and 8.5 (95% CI: 7.8-9.2) in the midwest; but 11.8 (95% CI: 10.8-12.8) in the northeast and 12.9 (95% CI: 12.0-13.9) in the south. In multivariate analyses, including sex, age, any stroke diagnosis, ethnicity (African-American and white), and region, the ethnic and regional differences persisted. The substantially higher gastrostomy placement rate among older hospitalized African Americans in the United States appears to be independent of geographical variation. PMID- 15540888 TI - Esophageal impacted dentures. AB - OBJECTIVES: This study aims to highlight the problems associated with impacted acrylic dentures and proffers advice to check them. PATIENTS AND METHODS: Retrospective review of all cases of impacted acrylic dentures over a 16-year period. RESULTS: Twenty-two adults had impacted esophageal acrylic dentures of which 16 (72.7%) and six (27.3%) were males and females, respectively (M:F ratio = 2.7:1) with age range 23-77 years. Fourteen patients (63.6%) had worn their dentures for more than 10 years without check-up, and 54.5% presented within 48 hours of impaction. The common symptoms in all the patients were difficulty with swallowing, throat pain and discomfort, followed by tenderness in the neck in 15 (68.2%). Dentures were extracted through esophagoscopy (17 cases) and cervical (three cases) esophagotomy, respectively. Observed complications included pulmonary edema in one and esophageal perforation in five patients. CONCLUSION: Endoscopic extraction of dentures carries a high risk of perforation. Extraction of an impacted denture via esophagoscopy can be undertaken under direct vision and in an ideal situation with judicious use of the Shears forceps. In the absence of these, the safest option is an esophagotomy. Proper treatment planning in the fabrication of dentures with incorporation of radiopaque materials in the dental resins and adequate postdenture delivery instructions are necessary as preventive measures. PMID- 15540889 TI - Diversity and professional excellence. AB - There are compelling moral and practical reasons why the memberships in professions should mirror the populations that they serve. In order to address this general issue more exactly, this essay will confine itself to a particular case as an emblem for the general point. The particular case is the profession of orthopedic surgeons in the United States. From an examination of this specific case, it is hoped that more general issues of racial, ethnic, and gender fairness within the professions will also be addressed. PMID- 15540890 TI - Men who have sex with men: answers about HIV vaccine research. PMID- 15540892 TI - Impact factor misleading, citing all references. PMID- 15540891 TI - Malignant melanoma in a black child: predisposing precursors and management. AB - Malignant melanoma (MM) remains a pediatric rarity world-wide, but perhaps more so in black Africans. To the best of our knowledge, the current report of MM in a two-and-a-half-year-old Nigerian who had a pre-existing congenital giant hairy nevus is probably the first (in an accessible literature) in a black African child. Primary neoplastic transformation and metastatic spread were suggested by the appearance of multiple swellings over the "garment" precursor nevus at the posterior trunk, multiple ipsilateral axillary nodal enlargement, and fresh occipital swellings postadmission. Smaller-sized hyperpigmented lesions with irregular, nonlobulated, and frequently hairy surfaces were also discernible over the upper and lower extremities, but the face, anterior trunk, and mucosal surfaces were relatively spared. A diagnosis of MM was confirmed by the subsequent histopathologic findings from the fine-needle aspirate and biopsy specimens. Chemotherapy was initiated but was truncated shortly after by parent pressured discharge. Despite the rarity of MM in a tropical African setting where management options are few, the current case underscores the need for a high clinical index of diagnostic suspicion, an early pursuit of investigative confirmation, and prophylactic excision in children with the predisposing skin lesions, like congenital giant hairy nevus. An expounded discourse of the possible precursors and management options of MM is provided. We emphasize the need for institutional cost subsidy for anticancer care in tropical children. PMID- 15540893 TI - Bcl-2-related genes in lymphoid neoplasia. AB - The proto-oncogene BCL-2 was discovered with the cloning of the t(14;18) chromosomal translocation responsible for human follicular lymphoma. Since then other members of the Bcl-2 family of cell death regulators have been identified and their roles in cell death, normal lymphoid development, and lymphoid neoplasia have been characterized. Bcl-2 family members are important in tumor initiation, progression, and response to chemotherapy, and altered expression levels of various members serve as prognostic markers in many lymphoid malignancies. There are promising cancer therapeutics now targeted at members of the Bcl-2 family. PMID- 15540894 TI - Cytokine-mediated cell survival. AB - Pathways through which signals emanating from cytokine receptors support cell survival have long been a focus of intensive research. For Baf-3, a murine interleukin 3-dependent cell line, the 2 distinct pathways involved are JAK/STATs/Bcl-xL and Ras/PI3-K. The latter is indispensable for long-term cell survival through down-regulation of Bim, a BH3-only cell death activator of the Bcl-2 superfamily. Thus, Bim is likely to be a key factor for cytokine-initiated regulation of cell survival in both hematopoietic cells and neuronal cells. Cytokines (like neurotrophic factors) regulate Bim expression at at least 3 levels: (1) at the messenger RNA (mRNA) level through transcriptional regulation and possibly through mRNA stability, (2) at the protein level through proteasome dependent regulation of protein degradation, and (3) by affecting subcellular localization through regulation of the potential to bind to the dynein motor complex. Bim function may be regulated in different ways in certain situations such that the relative importance of these 3 mechanisms may differ among cell types. For hematopoiesis, mRNA regulation seems to be the most important. Bim is also implicated in leukemogenesis caused by the Bcr-Abl chimeric tyrosine kinase and constitutively active mutants of receptor tyrosine kinases. PMID- 15540895 TI - Nuclear factor kappaB activation and antiapoptosis in mucosa-associated lymphoid tissue lymphoma. AB - Three distinct chromosomal translocations, t(11;18), t(1;14), and t(14;18), involving the API2-MALT1 fusion protein, BCL10, and MALT1 have been convincingly implicated in the pathogenesis of mucosa-associated lymphoid tissue (MALT) lymphomas. Recent genetic and biochemical studies have indicated that BCL10 and MALT1 form a physical and functional complex and are both essential for nuclear factor kappaB (NF-kappaB) activation by antigen receptor stimulation in lymphocytes. API2-MALT1 can bypass the BCL10/MALT1 pathway linking to NF-kappaB activation, thereby inducing antigen receptor-independent events of lymphocytes. BCL10/MALT1- and API2-MALT1-induced NF-kappaB activation can be assumed to be able to contribute to antiapoptosis, probably through NF-kappaB-mediated up regulation of several apoptotic inhibitor genes. We also have provided direct evidence that API2-MALT1 can exert an antiapoptotic effect, in part through its direct interaction with apoptotic regulators. We therefore hypothesize that the antiapoptotic effect of API2-MALT1 may be mediated by the interaction with apoptotic regulators as well as by the up-regulation of apoptotic inhibitor genes. Finally, we hope that further studies will stimulate research leading to the development of therapeutic drugs that specifically inhibit the antigen receptor-stimulated NF-kappaB activation pathway. Such drugs should be useful for interfering with inappropriate proliferation of lymphocytes associated with inflammatory and neoplastic disorders, including MALT lymphomas. PMID- 15540896 TI - Apoptosis of multiple myeloma. AB - Multiple myeloma (MM) is a malignancy of terminally differentiated plasma cells. MM cells localize to the bone marrow, where cell adhesion-mediated autocrine or paracrine activation of various cytokines, such as interleukin 6, insulin-like growth factor 1, and interferon alpha, results in their accumulation mainly because of loss of critical apoptotic controls. Resistance to apoptosis, a genetically regulated cell death process, may play a critical role in both pathogenesis and resistance to treatment of MM. Abnormalities in regulation and execution of apoptosis can contribute to tumor initiation, progression, as well as to tumor resistance to various therapeutic agents. Apoptosis is executed via 2 main pathways that lead to activation of caspases: the death receptor (extrinsic) pathway and the mitochondrial (intrinsic) pathway. Ionizing radiation and chemotherapeutic agents act primarily through the intrinsic pathway, in which mitochondria play the central role. Various therapeutic modalities that are effective in MM modulate levels of the proapoptotic and antiapoptotic Bcl-2 family of proteins and of inhibitors of apoptosis, expression of which is primarily regulated by p53, nuclear factor KB, and STAT (signal transducers and activators of transcription) factors. This review focuses on the key concepts and some of the most recent studies of signaling pathways regulated in MM and summarizes what is known about the clinical role of these pathways. PMID- 15540898 TI - Gene expression profiling of human erythroid progenitors by micro-serial analysis of gene expression. AB - We compared the expression profiles of highly purified human CD34+ cells and erythroid progenitor cells by micro-serial analysis of gene expression (microSAGE). Human CD34+ cells were purified from granulocyte colony-stimulating factor-mobilized blood stem cells, and erythroid progenitors were obtained by cultivating these cells in the presence of stem cell factor, interleukin 3, and erythropoietin. Our 10,202 SAGE tags allowed us to identify 1354 different transcripts appearing more than once. Erythroid progenitor cells showed increased expression of LRBA, EEF1A1, HSPCA, PILRB, RANBP1, NACA, and SMURF. Overexpression of HSPCA was confirmed by real-time polymerase chain reaction analysis. MicroSAGE revealed an unexpected preferential expression of several genes in erythroid progenitor cells in addition to the known functional genes, including hemoglobins. Our results provide reference data for future studies of gene expression in various hematopoietic disorders, including myelodysplastic syndrome and leukemia. PMID- 15540897 TI - Survivin and leukemia. AB - Survivin is a unique member of the inhibitor-of-apoptosis family. Survivin plays a role in the proliferation and survival of normal hematopoietic cells. Survivin expression is aberrantly enhanced in most cancers and hematopoietic malignancies. Survivin is an attractive therapeutic target, and various approaches to targeting survivin have been tested in vitro and in vivo. We review here progress in survivin research. PMID- 15540899 TI - Humanized anti-interleukin 6 receptor antibody induced long-term remission in a patient with life-threatening refractory autoimmune hemolytic anemia. AB - Refractory autoimmune hemolytic anemia (AIHA) is associated with considerable rates of mortality. Interleukin 6 (IL-6) has been reported to play a role in the pathogenesis of AIHA. This report describes a patient with AIHA who was successfully treated with a humanized anti-human IL-6 receptor (IL-6R) monoclonal antibody (MoAb). He had experienced life-threatening AIHA and had received conventional therapy with corticosteroids, azathioprine, cyclophosphamide, cyclosporin A, melphalan, plasma exchange, and irradiation to his spleen. However, the patient's symptoms and laboratory data did not show a sufficient improvement. Because his serum IL-6 level was elevated, we attempted to block IL 6 signaling by using a humanized anti-IL-6R MoAb, MRA. With 8 mg/kg of MRA administration every 2 weeks, the serum hemoglobin level gradually increased and normalized within 4 months. After 2 years of MRA treatment, the disease activity was well controlled without adverse reactions. Anti-IL-6R MoAb can be a novel and effective therapeutic agent for AIHA. PMID- 15540900 TI - Successful treatment with cyclosporine and high-dose gamma immunoglobulin for persistent parvovirus B19 infection in a patient with refractory autoimmune hemolytic anemia. AB - We describe a patient with persistent pure red cell aplasia due to human parvovirus B19 (HPVB19) infection during immunosuppressive therapy for refractory autoimmune hemolytic anemia (AIHA). The patient had been given corticosteroid (CS) and/or azathioprine for AIHA. During the course of treatment, reticulocyte count and hemoglobin levels decreased suddenly. Bone marrow aspirate showed erythroid lineage-specific aplasia with a few giant proerythroblasts, suggesting the presence of HPVB19 infection. The diagnosis of aplastic crisis due to HPVB19 infection was based on positive test results by polymerase chain reaction for HPVB19 immunoglobulin M (IgM) antibody and B19 DNA. Although splenectomy followed by administration of high-dose gamma globulin (HDIG) and plasma exchange were performed, the crisis and hemolysis recurred. Aplastic crises occurred several times when the B19 IgG result became negative and the CD4+ lymphocyte count was less than 300/microL. The patient showed complete recovery from anemia after CS was switched to cyclosporin A (CsA) and intermittent administration of HDIG. The result for B19 IgG antibody was continuously positive, and the DNA result became negative after these treatments. The results in this case indicated that concomitant administration of CsA and intermittent administration of HDIG can lead to cure of chronic anemia due to HPVB19 infection in patients with refractory AIHA. PMID- 15540901 TI - Clinical and oncologic implications in epigenetic down-regulation of CD26/dipeptidyl peptidase IV in adult T-cell leukemia cells. AB - CD26/dipeptidyl peptidase IV (DPPIV), a T-cell-activation antigen, is a 110-kD type II surface glycoprotein expressed on various types of normal cells. CD26/DPPIV is considered a multifunction housekeeping protein. Malignant cells often show altered CD26/DPPIV expression or no CD26/DPPIV expression, thus suggesting a useful marker for assessing some T-cell malignancies. In this study, cell surface protein and messenger RNA expression profiles for CD26/DPPIV were examined in 49 patients with adult T-cell leukemia (ATL), 10 carriers of human T lymphotropic virus I (HTLV-I), and 4 HTLV-I-infected cell lines to assess the utility of CD26/DPPIV expression as a useful molecular marker for ATL pathology. In contrast to normal lymphocytes, ATL cells and HTLV-I-infected cell lines apparently down-regulated or completely lost the CD26/DPPIV antigen. Furthermore, the positive rate and antigen density for CD26/DPPIV in ATL cells gradually declined along with the advancement of ATL stage. Analysis of genomic DNA and the CD26/DPPIV transcript showed that CD26- ATL cells possessed faintly detected transcripts of the gene that were aberrantly methylated at the CpG islands within the promoter region in parallel with the advancement of ATL, a finding supported by a rescue experiment for transcript reexpression using 5-azacytidine as demethylation agent. Moreover, there was no relationship between loss of CD26/DPPIV and HTLV-I tax expression. These results indicate that ATL cells down regulate CD26 antigens by means of epigenetic machinery and that this antigen abnormality is a useful molecular marker for the pathology of ATL. PMID- 15540902 TI - Efficacy and safety of imatinib mesylate for patients in the first chronic phase of chronic myeloid leukemia: results of a Japanese phase II clinical study. AB - Imatinib mesylate is a relatively new drug that targets the BCR-ABL chimeric protein, the molecular basis of chronic myeloid leukemia (CML). A phase II clinical trial in 39 Japanese patients in the first chronic phase of CML was conducted with imatinib mesylate at a dose of 400 mg/day. Hematologic complete response was obtained in 92.3% of the patients, complete cytogenetic response (CR) was obtained in 43.6%, and major partial CR was obtained in 20.5% of the patients. Although 29 of 39 patients required an adjustment of dosing because of grade 3 or 4 adverse events, most of the events were reversible, and 25 of the 29 patients were able to resume therapy. Between day 15 and day 35, grade 3 or 4 neutropenia and/or leukocytopenia occurred in 13 patients, and grade 3 thrombocytopenia occurred in 5 patients. Overall, nonhematologic grade 3 adverse events occurred in 28.2% of the patients. These data support the use of imatinib mesylate as the treatment of choice for chronic-phase CML patients. PMID- 15540904 TI - Aleukemic leukemia cutis in a patient with Philadelphia chromosome-positive biphenotypic leukemia. AB - Aleukemic leukemia cutis is a rare condition characterized by the invasion of leukemic blasts into the skin before their appearance in the peripheral blood. Leukemia cutis usually occurs in patients with myeloid leukemia, especially the myelomonocytic and monocytic types of acute myeloblastic leukemia. We describe the case of a 62-year-old woman with aleukemic leukemia cutis who developed Philadelphia-positive acute leukemia 1 month after skin involvement. Leukemic cells expressed both myeloid and B-cell lineage surface markers, and monoclonal rearrangement of the immunoglobulin heavy chain was detected by Southern blot analysis. This report is the first of a case of aleukemic leukemia cutis preceding Philadelphia-positive biphenotypic leukemia. PMID- 15540905 TI - In vitro generation of functional dendritic cells from human umbilical cord blood CD34+ cells by a 2-step culture method. AB - Dendritic cells (DCs) are the most potent antigen-presenting cells in terms of initiating primary T-cell-dependent immune responses. We devised a 2-step culture method for obtaining sufficient numbers of functional DCs from umbilical cord blood (CB) CD34+ cells. In the first step, CB CD34+ cells were expanded by stimulation with early-acting cytokines such as stem cell factor (SCF), flt3 ligand (FL), and thrombopoietin (TPO) to amplify the hematopoietic progenitor cells. In the second step, granulocyte-macrophage colony-stimulating factor and interleukin 4 were added, and incubation was continued for another 5 days to induce differentiation of the expanded cells into DCs. During the first step of culturing with TPO, SCF, and FL, the total numbers of nucleated cells gradually increased, peaking at 4 weeks (245.3-fold). During the second step, expression of CD1a, CD83, and CD86 increased. Electron microscopic findings showed that these cells had cytosolic expansion to form dendrites and major histocompatibility complex class II compartments, which are characteristic of DCs. Functional analyses revealed that these cells had phagocytic activity and were capable of stimulating allogeneic T-cells in vitro. PMID- 15540903 TI - Durable response but prolonged cytopenia after cladribine treatment in relapsed patients with indolent non-Hodgkin's lymphomas: results of a Japanese phase II study. AB - We conducted a phase II study to evaluate the efficacy and safety of cladribine (2-chlorodeoxyadenosine [2-CdA]) for patients with refractory or relapsed indolent B-cell lymphoma or mycosis fungoides. Forty-five patients were enrolled, and 43 patients, including 34 with follicular lymphoma, were eligible. 2-CdA was given by continuous intravenous infusion at a dose of 0.09 mg/kg daily for 7 consecutive days, and this schedule was repeated every 4 weeks up to a maximum of 6 cycles. The overall and complete response rates were 58.1% (25/43; 90% confidence interval, 44.5%-70.9%) and 14.0% (6/43), respectively. The disease progression-free proportions of all 43 eligible and all 25 responding patients at 2 years were 30.3% and 48.1%, respectively. Neutropenia and thrombocytopenia of grade 3 or 4 were observed in 53.3% and 37.8% of patients, respectively, with prolonged cytopenia observed in patients with increased numbers of treatment cycles. Nonhematologic toxicities of grade 3 or greater included diarrhea, arrhythmia, malaise, and gastrointestinal bleeding in 1 patient each, an increase in glutamic-pyruvic transaminase level in 2 patients, and infection in 5 patients. Two treatment-related deaths were observed. Four patients developed myelodysplastic syndrome (MDS) at 13 months to 2 years after completion of the 2 CdA treatments. 2-CdA is an active agent with acceptable toxicity for refractory or relapsed indolent lymphoma; however, prolonged myelosuppression and the potential development of MDS should be carefully monitored. PMID- 15540906 TI - Successful unrelated cord blood transplantation using a reduced-intensity conditioning regimen in a 6-month-old infant with congenital neutropenia complicated by severe pneumonia. AB - Here we report the first successful unrelated cord blood transplantation (CBT) using reduced-intensity conditioning for the 'treatment of congenital neutropenia in a 6-month-old infant with complications of severe pneumonia probably due to Staphylococcus aureus infection. Because the patient showed no response to treatment with granulocyte colony-stimulating factor and had a cytogenetic aberration, unrelated CBT with an HLA-DRB1 genotypic mismatch was performed. The number of infused cells was 15 x 10(7)/kg. The preparative regimen was fludarabine, cyclophosphamide, and 6 Gy of total body irradiation. Teicoplanin was administered for bacterial pneumonia. Neutrophil engraftment was achieved on day 41 and was followed by clinical improvement. The patient gradually caught up on growth and development after the CBT. Unrelated CBT using a reduced-intensity conditioning regimen may be an effective treatment for congenital neutropenia. PMID- 15540907 TI - Successful treatment of tacrolimus (FK506)-related leukoencephalopathy with cerebral hemorrhage in a patient who underwent nonmyeloablative stem cell transplantation. AB - A 46-year-old woman with Hodgkin's disease who underwent nonmyeloablative allogeneic stem cell transplantation developed cortical blindness, seizures, and left hemiparesis on day 100 while receiving tacrolimus (FK506) and prednisone for the treatment of graft-versus-host disease (GVHD). Magnetic resonance imaging revealed multiple changes, mainly in the bilateral occipital lobes, suggesting FK506-related leukoencephalopathy. These abnormalities improved after discontinuation of FK506. However, 3 days after the episode, cerebral hemorrhage in the left occipital lobe with perforation to the left subdural space occurred. Although FK506-induced leukoencephalopathy with cerebral hemorrhage is considered the more severe form of such leukoencephalopathy, the patient's neurological symptoms almost completely resolved and radiographic findings improved after discontinuation of FK506, tapering of methylprednisolone, and initiation of mycophenolate mofetil. FK506-related leukoencephalopathy is a rare complication after allogeneic stem cell transplantation. Although the symptoms usually subside after discontinuation of FK506, therapeutic intervention in many cases may result in severe complications, including GVHD and vascular disease. We consider it important to use immunosuppressive agents without vascular endothelial toxicity for preventing the development of fatal GVHD after discontinuation of FK506. PMID- 15540908 TI - Genetic polymorphism of the CYP2C9 subfamily of 3 different races in warfarin maintenance dose. PMID- 15540909 TI - Impact of temporal lobe surgery. PMID- 15540910 TI - White matter fiber dissection of the optic radiations of the temporal lobe and implications for surgical approaches to the temporal horn. AB - OBJECT: The aim of this anatomical study was to define more fully the three dimensional (3D) relationships between the optic radiations and the temporal horn and superficial anatomy of the temporal lobe by using the Klingler white matter fiber dissection technique. These findings were correlated with established surgical trajectories to the temporal horn. Such surgical trajectories have implications for amygdalohippocampectomy and other procedures that involve entering the temporal horn for the resection of tumors or vascular lesions. METHODS: Ten human cadaveric hemispheres were prepared with several cycles of freezing and thawing by using a modification of the method described by Klingler. Wooden spatulas were used to strip away the deeper layers of white matter progressively in a lateromedial direction, and various association, projection, and commissural fibers were demonstrated. As the dissection progressed, photographs of each progressive layer were obtained. Special attention was given to the optic radiation and to the sagittal stratum of which the optic radiation is a part. The trajectories of fibers in the optic radiation were specifically studied in relation to the lateral, medial, superior, and inferior walls of the temporal horn as well as to the superficial anatomy of the temporal lobe. In three of the hemispheres coronal sections were made so that the relationship between the optic radiation and the temporal horn could be studied more fully. In all 10 hemispheres that were dissected the following observations were made. 1) The optic radiation covered the entire lateral aspect of the temporal horn as it extends to the occipital horn. 2) The anterior tip of the temporal horn was covered by the anterior optic radiation along its lateral half. 3) The entire medial wall of the temporal horn was free from optic radiation fibers, except at the level at which these fibers arise from the lateral geniculate body to ascend over the roof of the temporal horn. 4) The superior wall of the temporal horn was covered by optic radiation fibers. 5) The entire inferior wall of the temporal horn was free from optic radiation fibers anterior to the level of the lateral geniculate body. CONCLUSIONS: Fiber dissections of the temporal lobe and horn demonstrated the complex 3D relationships between the optic radiations and the temporal horn and superficial anatomy of the temporal lobe. Based on the results of this study, the authors define two anatomical surgical trajectories to the temporal horn that would avoid the optic radiations. The first of these involves a transsylvian anterior medial approach and the second a pure inferior trajectory through a fusiform gyrus. Lateral approaches to the temporal horn through the superior and middle gyri, based on the authors' findings, would traverse the optic radiations. PMID- 15540911 TI - Anatomical landmarks for hemispherotomy and their clinical application. AB - OBJECT: The authors introduce the surgical concept of the central core of a hemisphere, from which anatomical structures are disconnected during most current hemispherotomy techniques. They also propose key anatomical landmarks for hemispherotomies that can be used to disconnect the hemisphere from its lateral surface around the insula, through the lateral ventricle toward the midline. METHODS: This anatomical study was performed in five adult cadaveric heads following perfusion of the cerebral arteries and veins with colored latex. Anatomical landmarks were used in five hemispheric deafferentations. The central core of a hemisphere consists of extreme, external, and internal capsules; claustrum; lentiform and caudate nuclei; and thalamus. Externally, this core is covered by the insula and surrounded by the fornix, choroid plexus, and lateral ventricle. During most hemispherotomies, the surgeon reaches the lateral ventricle through the frontoparietal opercula or temporal lobe; removes the mesial temporal structures; and disconnects the frontal lobe ahead, the parietal and occipital lobes behind, and the intraventricular fibers of the corpus callosum above the central core. After a temporal lobectomy, the landmarks include the choroid plexus and posterior/ascending portion of the tentorium to disconnect the parietal and occipital lobes, the callosal sulcus or distal anterior cerebral artery (ACA) to sever the intraventricular fibers of the corpus callosum, and the head of the caudate nucleus and ACA to detach the frontal lobe. CONCLUSIONS: These landmarks can be used in any hemispherotomy during which a cerebral hemisphere is disconnected from its lateral surface. Furthermore, they can be used to perform any resection around the central core of the hemisphere and the tentorial incisura. PMID- 15540912 TI - Prophylactic administration of ceftriaxone for the prevention of meningitis after traumatic pneumocephalus: results of a clinical trial. AB - OBJECT: The purpose of this study was to compare the efficacy of the prophylactic use of ceftriaxone for the prevention of meningitis in patients with acute traumatic pneumocephalus. METHODS: In this prospective, single-institution, randomized clinical trial, 109 patients with mild head injury and traumatic pneumocephalus were randomly assigned to receive or not receive an antibiotic medication (ceftriaxone, 1 g given twice a day) until occurrence of meningitis or at least 5 days after trauma. The patients were followed up for 1 month posttrauma. The 109 patients were divided into two groups: 53 were assigned to the prophylactic antibiotics therapy group and 56 to the control group. The overall rate of meningitis was 20.1% and the rates of meningitis in the two groups were not significantly different. The results were the same when adjusted for the patient's Glasgow Coma Scale score, sex, and age, as well as for an intradural location of air, air volume, presence of cerebrospinal fluid (CSF) rhinorrhea or CSF otorrhea, radiological sign of a skull base fracture, or intracranial hemorrhage. CONCLUSIONS: The results of this study do not substantiate the efficacy of ceftriaxone used in the prevention of meningitis in patients with traumatic pneumocephalus after mild head injury or in any specific subgroup of these patients. Cerebrospinal fluid rhinorrhea and intracranial hemorrhage may be considered primary risk factors for the development of meningitis in patients with posttraumatic pneumocephalus and, in the absence of these symptoms, intradural location of air and air volume greater than 10 ml may be considered secondary risk factors. Further studies in this area are warranted. PMID- 15540913 TI - Continuous-flow shunt for treatment of hydrocephalus due to lesions of the posterior fossa. AB - OBJECT: Management of hydrocephalus caused by expansive lesions of the posterior fossa is complicated by two main drawbacks of shunt devices: sudden decompression and overdrainage. The ventriculoperitoneal (VP) continuous-flow (CF) shunt is characterized by a peritoneal catheter with an internal diameter of 0.51 mm that promotes continuous drainage of cerebrospinal fluid (CSF) at its production rate. The authors have previously demonstrated in adult patients with hydrocephalus that sudden decompression and overdrainage are absent when this shunt is used; here they report the findings of a prospective study in which the goal was to test the performance of this shunt in patients with severe hydrocephalus due to lesions of the posterior fossa. METHODS: During a 5-year period, 103 patients with severe hydrocephalus caused by lesions of the posterior fossa were treated by placement of a VP shunt. In 53 of these patients (control group) a shunt and valve system was surgically implanted and in 50 patients a CF shunt was implanted. All patients were followed up for a minimum of 2 years after surgery. Shunt revision or change was necessary in 21 patients (40%) with conventional shunts and in four patients (8%) with the CF shunt (p < 0.003). Signs of overdrainage were observed in 18 patients (34%) in the control group, four of whom had ascending transtentorial herniation; this complication was not seen in patients with the CF shunt. CONCLUSIONS: The CF shunt had a low rate of dysfunction and an absence of complications caused by overdrainage, which were frequently associated with the control shunts. The hydrodynamic properties of the CF shunt make it effective, even in severe cases of hydrocephalus caused by lesions of the posterior fossa. PMID- 15540914 TI - Transradial approach for neuroendovascular surgery of intracranial vascular lesions. AB - OBJECT: The authors present their experience in performing a transradial approach for neuroendovascular surgery of intracranial vascular lesions when a transfemoral approach was unfavorable. METHODS: Eight patients ranging in age from 52 to 88 years underwent a total of nine neuroendovascular procedures for intracranial vascular lesions. A transradial approach was used in all patients. The patients had previously undergone a transfemoral approach for the endovascular intervention, but that procedure was unsuccessful. Five patients had intracranial basilar artery (BA) aneurysms, one patient had symptomatic BA stenosis, one patient had a dural arteriovenous fistula in the posterior fossa, and one patient had a high-flow arteriovenous malformation in the frontal lobe. In each case, a transradial approach achieved a stable platform that allowed intracranial microcatheterization for neuroendovascular intervention. None of the patients experienced complications attributed to the transradial artery approach. CONCLUSIONS: During neuroendovascular surgery for the treatment of intracranial lesions, the transradial approach is a viable alternative if the transfemoral approach is unfavorable. This series represents the first known description of neuroendovascular surgery for intracranial lesions via a transradial approach. PMID- 15540915 TI - Contralateral motor rootlets and ipsilateral nerve transfers in brachial plexus reconstruction. AB - OBJECT: The goal of this study was to evaluate outcomes in patients with brachial plexus avulsion injuries who underwent contralateral motor rootlet and ipsilateral nerve transfers to reconstruct shoulder abduction/external rotation and elbow flexion. METHODS: Within 6 months after the injury, 24 patients with a mean age of 21 years underwent surgery in which the contralateral C-7 motor rootlet was transferred to the suprascapular nerve by using sural nerve grafts. The biceps motor branch or the musculocutaneous nerve was repaired either by an ulnar nerve fascicular transfer or by transfer of the 11th cranial nerve or the phrenic nerve. The mean recovery in abduction was 90 degrees and 92 degrees in external rotation. In cases of total palsy, only two patients recovered external rotation and in those cases mean external rotation was 70 degrees. Elbow flexion was achieved in all cases. In cases of ulnar nerve transfer, the muscle scores were M5 in one patient, M4 in six patients, and M3+ in five patients. Elbow flexion repair involving the use of the 11th cranial nerve resulted in a score of M3+ in five patients and M4 in two patients. After surgery involving the phrenic nerve, two patients received a score of M3 + and two a score of M4. Results were clearly better in patients with partial lesions and in those who were shorter than 170 cm (p < 0.01). The length of the graft used in motor rootlet transfers affected only the recovery of external rotation. There was no permanent injury at the donor sites. CONCLUSIONS: Motor rootlet transfer represents a reliable and potent neurotizer that allows the reconstruction of abduction and external rotation in partial injuries. PMID- 15540916 TI - Volumetric thermal devascularization of large meningiomas. AB - OBJECT: Controlling hemorrhage is crucial in the safe and efficient removal of large meningiomas. Intravascular embolization is not always a satisfactory means of accomplishing this goal because of the procedure's hemostatic effect and risk of complications. The authors in this study used a volumetric thermal ablation technique incorporating radiofrequency energy, image guidance, and local temperature control to devascularize tumor tissue. METHODS: Five patients with large meningiomas were treated. The target and orientation of the radiofrequency thermal ablation (RFTA) were simulated preoperatively to maximize devascularization of the lesion without thermal injury to adjacent critical structures. Image fusion, three-dimensional reconstruction, and image-guided methods provided for optimized trajectories and targets for insertion of the RFTA needle. During ablation, local temperatures of the tissue being cauterized were monitored continuously to limit the ablated lesion to within the target volume. The effects of devascularization and the softening of the tumor parenchyma facilitated lesion removal. The intracranial ablated meningioma changed into necrotic tissue and shrank within a few months. Histopathological examination of the ablated lesion revealed sharply demarcated coagulation necrosis. CONCLUSIONS: Volumetric thermal devascularization can be applied safely in the treatment of large meningiomas to facilitate surgical manipulation of the lesion as well as to reduce its size palliatively. The procedure's usefulness should be studied further in a larger number of cases with different tumor characteristics. PMID- 15540917 TI - Writing-specific sites in frontal areas: a cortical stimulation study. AB - OBJECT: The aim of this study was to determine whether cortical areas involved in the writing process are associated with reading or naming areas in patients undergoing surgery for brain tumors in frontal areas. This study was undertaken to spare all language areas found in patients during surgery. METHODS: Fourteen patients (eight women and six men [mean age 47 years] of whom 12 were right handed, two left handed, 12 monolingual, and two bilingual) who harbored brain tumors in the left (11 patients) or right (three patients) frontal gyri or in rolandic areas, were tested by direct stimulation by using the awake surgery technique for direct brain mapping. Mapping of the frontal gyri was performed using naming, reading, and writing under dictation tasks in the appropriate language(s). Considerable individual variability in language organization among patients was observed. Interferences in writing were found during direct stimulation in the frontal gyri, in cortical sites common or not common to interferences in naming or reading. In dominant regions, patterns of writing dysfunctions were variable and included writing arrest, illegible script, letter omissions, and paragraphia. These dysfunctions were nonspecific (stimulation induced eye movements) in nondominant frontal regions and in rolandic gyri (hand contractions). In the same patient, different writing impairments could sometimes be observed during stimulation of different sites. As is the case for naming or reading interference sites, writing interference sites could be extremely localized (1 cm2 in diameter). In this group of patients, writing interference sites found in Broca areas were associated with other sites of language interference, whereas writing-specific interference sites were found twice in the dominant middle frontal gyrus. CONCLUSIONS: In this series, we found that writing interference sites could be detected by direct cortical stimulation in dominant inferior and middle frontal gyri regardless of whether they were associated with naming or reading interference sites. Writing disorders elicited by direct stimulation in the frontal lobes are varied and probably depend on the functional status of the stimulated cortical area. PMID- 15540918 TI - Transient focal cerebral ischemia-induced neurogenesis in the dentate gyrus of the adult mouse. AB - OBJECT: Throughout the life of a mammal, new neurons are produced each day from resident progenitor cells located in the hippocampal dentate gyrus (DG). The availability of transgenic and knockout mice enables the evaluation of specific molecular mediators of this phenomenon. To facilitate such studies the authors characterized the proliferation, survival, and maturation of progenitor cells in the DG of adult mice following transient focal cerebral ischemia. METHODS: Anesthesia was induced in adult C57BL/6 mice by administering halothane. The middle cerebral artery (MCA) was then occluded for 120 minutes by applying an endovascular suture. The marker used to detect the presence of proliferating cells, 5-bromodeoxyuridine (BrdU; 50 mg/kg) was administered intraperitoneally twice daily on Days 2 through 6 after the MCA occlusion. Cohorts of mice were killed on Days 7 and 21, after which their brains were sectioned and BrdU positive cells were detected using immunohistochemical analysis. The phenotype of the BrdU-positive cells was identified by fluorescent triple labeling by using antibodies specific for neuronal and astroglial markers together with anti-BrdU antibodies. The infarction was confirmed by applying cresyl violet staining. Compared with sham-operated control animals, there was a 4.6-fold (p < 0.05) increase in BrdU-positive cells in the ipsilateral DG at Day 7 postischemia. Twenty-one percent of the newly proliferated cells survived to Day 21 postischemia. At this time, the newly proliferated cells expressed the immature and mature neuron markers doublecortin and NeuN, respectively, but none expressed the astroglial marker glial fibrillary acidic protein. CONCLUSIONS: Focal ischemia induces neurogenesis in the DG of the mouse brain; this may be critical for postischemic brain repair. PMID- 15540919 TI - Peripheral nerve regeneration by transplantation of bone marrow stromal cell derived Schwann cells in adult rats. AB - OBJECT: Bone marrow stromal cells (BMSCs) can be induced to form Schwann cells by sequentially treating the cells with beta-mercaptoethanol and retinoic acid, followed by forskolin and neurotrophic factors including heregulin. In this study the authors made artificial grafts filled with BMSC-derived Schwann cells (BMSC DSCs) and transplanted them into the transected sciatic nerve in adult rats to evaluate the potential of BMSCs as a novel alternative method of peripheral nerve regeneration. METHODS: The BMSC-DSCs were suspended in Matrigel and transferred into hollow fibers (12 mm in length), which were transplanted into the transected sciatic nerve in adult Wistar rats. Six months after cell transplantation, electrophysiological evaluation and walking track analysis were performed. Results of these studies showed significant improvement in motor nerve conduction velocity and sciatic nerve functional index in the BMSC-DSC-transplanted group compared with the control group (Matrigel graft only). Immunohistochemical study data demonstrated that transplanted BMSCs labeled with retrovirus green fluorescent protein were positive for P0 and myelin-associated glycoprotein and had reconstructed nodes of Ranvier and remyelinated regenerated nerve axons. The number of regenerated axons in the axial section of the central portion of the graft was significantly greater in the transplanted group. Although BMSCs can differentiate into several types of cells, tumor formation did not occur 6 months after engraftment. CONCLUSIONS: Results in this study indicate that BMSC-DSCs have great potential to promote regeneration of peripheral nerves. The artificial graft made with BMSC-DSCs represents an alternative method for the difficult reconstruction of a long distance gap in a peripheral nerve. PMID- 15540920 TI - Atorvastatin reduction of intravascular thrombosis, increase in cerebral microvascular patency and integrity, and enhancement of spatial learning in rats subjected to traumatic brain injury. AB - OBJECT: Atorvastatin, a beta-hydroxy-beta-methylglutaryl coenzyme A reductase inhibitor, has pleiotropic effects, such as promoting angiogenesis, increasing fibrinolysis, and reducing inflammatory responses, and has shown promise in enhancing recovery in animals with traumatic brain injury (TBI) and stroke. The authors tested the effect of atorvastatin on vascular changes after TBI. METHODS: Male Wistar rats subjected to controlled cortical impact injury were perfused at different time points with fluorescein isothiocyanate (FITC)--conjugated dextran 1 minute before being killed. Spatial memory function had been measured using a Morris Water Maze test at various points before and after TBI. The temporal profile of intravascular thrombosis and vascular changes was measured on brain tissue sections by using a microcomputer imaging device and a laser confocal microscopy. The study revealed the following results. 1) Vessels in the lesion boundary zone and hippocampal CA3 region showed a variety of damage, morphological alterations, reduced perfusion, and intraluminal microthrombin formation. 2) Atorvastatin enhanced FITC-dextran perfusion of vessels and reduced intravascular coagulation. 3) Atorvastatin promoted the restoration of spatial memory function. CONCLUSIONS: These results indicated that atorvastatin warrants investigation as a potential therapeutic drug for TBI. PMID- 15540921 TI - Atorvastatin reduction of intracranial hematoma volume in rats subjected to controlled cortical impact. AB - OBJECT: Atorvastatin, a beta-hydroxy-beta-methylglutaryl coenzyme A reductase inhibitor, has pleiotropic effects such as improving thrombogenic profile, promoting angiogenesis, and reducing inflammatory responses and has shown promise in enhancing neurological functional improvement and promoting neuroplasticity in animal models of traumatic brain injury (TBI), stroke, and intracranial hemorrhage. The authors tested the effect of atorvastatin on intracranial hematoma after TBI. METHODS: Male Wistar rats were subjected to controlled cortical impact, and atorvastatin (1 mg/kg) was orally administered 1 day after TBI and daily for 7 days thereafter. Rats were killed at 1, 8, and 15 days post TBI. The temporal profile of intraparenchymal hematoma was measured on brain tissue sections by using a MicroComputer Imaging Device and light microscopy. CONCLUSIONS: Data in this study showed that intraparenchymal and intraventricular hemorrhages are present 1 day after TBI and are absorbed at 15 days after TBI. Furthermore, atorvastatin reduces the volume of intracranial hematoma 8 days after TBI. PMID- 15540923 TI - Endoscopic anatomy of the chordae willisii in the superior sagittal sinus. AB - OBJECT: Chordae willisii are structures located in the lumen of the superior sagittal sinus (SSS). It is thought that they act as flow-improving structures within the sinuses. There are few anatomical descriptions of chordae willisii, and all previous observations were performed through standard anatomical dissections. The purpose of this study was to visualize and describe structural and topographical features of the chordae willisii with the aid of rigid endoscopy. METHODS: Twenty-five SSSs obtained from fresh human cadavers during autopsies were the material for this study. Specimens were flushed with tap water to remove clots. Bridging veins emptying into the sinus were ligated, and continuous flow of a saline solution through the sinus in a physiological direction was achieved by connecting the sinus to an irrigating system. Rigid endoscopes of different diameters (2.7-4.5 mm) and optic (0 and 30 degrees) were inserted into the lumen of the sinus. The endoscope was connected to a digital camera and a video system to allow for recording of the observed structures. Finally, the sinuses were opened and the chordae willisii were dissected using standard anatomical methods. The chordae willisii were observed in all examined specimens. Three different types of the cords were found: lamellar, trabecular, and valvelike types. The most common type was the valvelike (mixed) one, which comprised 45.1% of all cords. The chordae willisii were most commonly observed in the parietooccipital region of the SSS. CONCLUSIONS: Witout disturbing any structural relationships, the use of endoscopy allowed visualization and description of intraluminal structures as they behaved physiologically. PMID- 15540922 TI - Invasive phenotype observed in 1,3-bis(2-chloroethyl)-1-nitrosourea-resistant sublines of 9L rat glioma cells: a tumor model mimicking a recurrent malignant glioma. AB - OBJECT: Chemotherapy is suspected of having an effect on the generation of phenotypical heterogeneity and the development of drug resistance in tumors. Recurrent gliomas feature drug resistance as well as greater invasive growth than original tumors. The authors investigated phenotypical changes in invasion observed in 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-resistant sublines of the 9L rat gliosarcoma. METHODS: Two established BCNU-resistant sublines, derived from 9L gliosarcoma cells by treating these cells with BCNU in vivo or in vitro, were used in the study. An in vitro examination confirmed the resistance of the cells to BCNU treatment. The cells were implanted into the striatum of Fisher 344 rats, and histological examinations were performed to compare the growth patterns of the resultant tumors. A new brain tumor model was established by implanting 9L 2 cells in Fisher 344 rats. The 9L-2 and BTRC-19 cells displayed a distinct increase in BCNU resistance compared with the 9L cells. Both BCNU-resistant sublines developed a tumor mass with invasive margins, which is not the case with 9L tumor models. The newly developed 9L-2 tumor model demonstrated 100% tumor uptake with consistent growth patterns. CONCLUSIONS: Cells that acquire drug resistance also demonstrated invasive growth. Because the 9L-2 and BTRC-19 cells were derived from 9L cells that had been treated with BCNU in vivo and in vitro, this change in phenotype was likely caused by the drug treatment, which may have implications for chemotherapy of gliomas. The tumor model that developed from the 9L-2 cells can be used as a model of a recurrent glioma, which features drug resistance and invasive growth. PMID- 15540924 TI - Chorioretinal atrophy in a patient with moyamoya disease. Case report. AB - Moyamoya disease is characterized by constrictions of segments of the internal carotid arteries (ICAs) and a resultant abnormal anastomotic network. In the literature, visual disturbances from cerebrovascular accidents in patients with moyamoya disease have been described, but very few reports of intraocular pathological conditions have been published. The authors describe a patient with moyamoya disease who presented with chorioretinal atrophy; an association between these two diseases has not previously been reported in the literature. Findings of a clinical ophthalmological evaluation and angiographic series are presented. During the fundic examination, evidence of chorioretinal atrophy was found in this patient. Choroidal vascular insufficiency was revealed by intravenous fluorescein angiography and occlusion of the ICAs proximal to the origin of the posterior communicating arteries by selective carotid and vertebral arteriography. The vertebrobasilar system provided anastomotic connections via the posterior communicating arteries. This is the first case report of chorioretinal atrophy associated with moyamoya disease. It is believed that the vasoocclusive effects of moyamoya disease may predispose the patient to atrophic changes in the peripheral retina. The development of an anastomotic network precludes the progression of this fundic anomaly. PMID- 15540925 TI - Complex visual hallucinations (Charles Bonnet syndrome) in visual field defects following cerebral surgery. Report of four cases. AB - The development of visual hallucinations after loss of vision is known as the Charles Bonnet syndrome. This phenomenon was first described in 1760 by Charles Bonnet and others during their observations of elderly patients with degeneration of the retina or cornea. To date a clear association between visual hallucinations and neurosurgical procedures has not been reported. Because of their clear demarcation, however, surgical lesions in the cerebrum offer a unique opportunity to determine the pathoanatomical aspects of visual hallucinations. During a 3-year period, 41 consecutive patients who acquired visual field defects after neurosurgery were examined for the occurrence of visual hallucination. Postoperatively, four of these patients experienced visual hallucinations. In two of them an upper quadrantanopia developed after the patients had undergone selective amygdalohippocampectomy. In the other two patients a complete hemianopia developed, in one case after resection of a parietal astrocytoma and in the other after resection of an occipital glioblastoma multiforme. The visual hallucinations were transient and gradually disappeared between 4 days and 6 months postoperatively. The patients were aware of the fact that their hallucinations were fictitious and displayed no psychosis. Electroencephalographic recordings were obtained in only two patients and epileptic discharges were found. Deafferentiation of cortical association areas may lead to the spontaneous generation of complex visual phenomena. In the present series this phenomenon occurred in approximately 10% of patients with postoperative visual field defects. In all four cases the central optic radiation was damaged between the lateral geniculate nucleus and the primary visual cortex. The complex nature of the visual hallucination indicates that they were generated in visual association areas. PMID- 15540926 TI - Association of an irregularly shaped anterior choroidal aneurysm with CREST syndrome. Case report. AB - The authors present the case of a 50-year-old woman with a history of CREST syndrome (calcinosis, Raynaud phenomenon, esophageal motility disorders, sclerodactyly, and telangiectasia), a variant of scleroderma, who was incidentally found to have an irregular intracranial aneurysm. The patient presented with migraine headaches. A magnetic resonance image of the brain obtained during the headache workup revealed a right posterior carotid artery wall aneurysm in the region of the anterior choroidal artery (AChA). On digital subtraction angiograms, the lesion measured 3.5 mm at its largest diameter. Because of the irregular shape of the aneurysm, the patient's relatively young age, and the potential for further aneurysm growth due to collagen disease, surgical clip application was recommended following a discussion of available treatment options. At surgery, the aneurysm was identified as bilobed and broad based, and the AChA was found to be associated with the aneurysm neck. Satisfactory clipping of the aneurysm was achieved with preservation of the parent vessels. An association of CREST syndrome with intracranial aneurysms has only been reported once before. This case is presented to draw attention to the possibility of a pathophysiological connection between CREST syndrome and intracranial aneurysms and to postulate a possible mechanism whereby this condition may result in aneurysm formation. The association of aneurysms with other pathological collagen-related conditions is well known, and literature relevant to a possible connection between CREST syndrome and aneurysms is reviewed and discussed. PMID- 15540927 TI - A carcinoid tumor mimicking an isolated intracranial meningioma. Case report. AB - This 79-year-old woman presented with progressively worsening dementia, abulia, flat affect, urinary incontinence, and profuse watery diarrhea. Results of computerized tomography and magnetic resonance studies indicated an extraaxial, dural-based mass compressing the right frontal lobe and consistent with a convexity meningioma. A right frontal craniotomy was performed and the dural based mass was resected. Histopathological features on immunostaining of the lesion were consistent with a carcinoid tumor (low-grade neuroendocrine carcinoma). Further evaluation revealed no primary carcinoid tumor in the foregut from which they typically originate. The authors concluded that this intracranial carcinoid tumor was the primary lesion despite its unusual location and that it should be included in the differential diagnosis of dural-based, extraaxial brain lesions. PMID- 15540928 TI - Monitoring of the lateral spread response in the endovascular treatment of a hemifacial spasm caused by an unruptured vertebral artery aneurysm. Case report. AB - The lateral spread response (LSR) is used in the electrophysiological diagnosis of a hemifacial spasm or for monitoring during microvascular decompression. The authors used LSRs for intraoperative monitoring during endovascular surgery in a rare case of vertebral artery (VA) aneurysm that caused intractable hemifacial spasm. A 49-year-old woman presented with a right hemifacial spasm that had persisted for 9 months. No other clinical symptom was observed. Vertebral artery angiography revealed a saccular aneurysm of the right VA. Magnetic resonance (MR) imaging demonstrated that the aneurysm was compressing the root exit zone of the right facial nerve. Endovascular treatment of the VA aneurysm was performed while monitoring the patient's LSRs. During occlusion of the VA at sites distal and proximal to the aneurysm, the LSRs temporarily disappeared and then reappeared with a higher amplitude than those measured preceding their disappearance. The hemifacial spasm alleviated gradually and disappeared completely 6 months after treatment. The LSRs changed in parallel with the improvement in the patient's hemifacial spasms and eventually disappeared. No recurrence of symptoms has been noticed as of 18 months postoperatively. This is the first report of the use of LSR monitoring during endovascular surgery for an intracranial aneurysm that causes hemifacial spasm. Intraoperative and postoperative changes in the LSRs provided useful information regarding the pathophysiology of hemifacial spasm. PMID- 15540929 TI - Preoperative stent placement for intradural vertebral artery stenosis from a rare xanthogranuloma. Case report. AB - In this report, the authors discuss a novel use of intradural vertebral artery (VA) stent placement to protect a tumor-encased vessel from injury during lesion resection. The tumor was a rare foramen magnum region xanthogranuloma and a component of Erdheim-Chester disease (ECD). This 64-year-old man presented with large masses encasing and compressing the intracranial segments of each VA. Preoperatively, a left VA stent was placed to protect the arterial wall during resection of the tumor. Histopathological study results on the subtotally resected mass were consistent with xanthogranuloma, a rare benign histiocytic tumor frequently occurring in patients with ECD. Further radiographic evaluation in the patient revealed an osteolytic lesion of the eleventh thoracic vertebra supporting the diagnosis of ECD disease. Based on this case study, the authors recommend the following: 1) tumor-encased vessels can be protected preoperatively by stent placement to assist with tumor debulking; and 2) patients diagnosed with a xanthogranuloma should be evaluated for multisystem involvement consistent with ECD. PMID- 15540930 TI - Hemorrhage from moyamoya-like vessels associated with a cerebral arteriovenous malformation. Case report. AB - The authors describe a case of subarachnoid hemorrhage from moyamoya-like vessels associated with an arteriovenous malformation (AVM) in a 44-year-old Hispanic man who presented with severe headache. The AVM was located in the left parietal lobe and the ipsilateral middle cerebral artery was occluded. Although the patient was initially neurologically intact, he began to experience neurological deficits from mild vasospasm, illustrating the sensitivity of the underperfused portion of brain surrounding an AVM. His neurological deficits improved with aggressive hydration and elevated blood pressure. After a 3-week period, the AVM was resected without complication and all of the patient's neurological deficits resolved. The authors review radiographic findings of this unique case. PMID- 15540931 TI - Pathophysiology of trigeminal neuralgia: new evidence from a trigeminal ganglion intraoperative microneurographic recording. Case report. AB - The origin of trigeminal neuralgia (TN) appears to be vascular compression of the trigeminal nerve at the root entry zone; however, the physiological mechanism of this disorder remains uncertain. The authors obtained intraoperative microneurographic recordings from trigeminal ganglion neurons in a patient with TN immediately before percutaneous radiofrequency-induced gangliolysis. Their findings are consistent with the idea that the pain of TN is generated, at least in part, by an abnormal discharge within the peripheral nervous system. PMID- 15540932 TI - Secretion of high-molecular-weight adrenocorticotropic hormone from a pituitary adenoma in a patient without Cushing stigmata. Case report. AB - The authors report a case in which a patient harbored a corticotroph macroadenoma that secreted biologically inactive high-molecular-weight adrenocorticotropic hormone (ACTH) as well as authentic ACTH 1-39. The secretion of the high molecular-weight ACTH was determined using gel chromatography. The authors believe that these two molecules competed with each other at the ACTH receptor and, thus, the bioactivity of ACTH 1-39 was masked and Cushing features were not manifested in the patient. This type of silent corticotroph adenoma may be categorized as a clinically nonfunctioning adenoma. Plasmas from patients with silent corticotroph adenomas, which are identified by positive immunohistochemical staining of ACTH, should be frozen, stored, and analyzed using gel chromatography to examine whether the tumors produce and secrete high molecular-weight ACTH. PMID- 15540933 TI - Resolution of pseudotumor cerebri after bariatric surgery for related obesity. Case report. AB - Two obese women, both of whom were 42 years of age, were examined for pseudotumor cerebri. Intracranial venography revealed increased pressure in the dural venous sinuses and the right atrium. The increased right atrial pressure was attributable to the patients' obesity. Both patients underwent bariatric surgery to achieve weight loss. Approximately 1 year later, a clinical evaluation showed that in both women the pseudotumor cerebri had resolved. Repeated measurements of dural venous pressure indicated that the patients' pressures had returned to normal. Obese patients with pseudotumor cerebri and stable visual symptoms are best treated with weight loss to avoid shunt placement or optic nerve sheath fenestration. PMID- 15540934 TI - The Brain Research Laboratory at the Cleveland Metropolitan General Hospital and case Western Reserve University. AB - In 1961 Dr. Robert White, fresh from the Mayo Clinic, established the Brain Research Laboratory (BRL) at the Cleveland Metropolitan General Hospital, Cleveland, Ohio, under the auspices of Case Western Reserve University. During a span of 15 years, he and his colleagues contributed significantly to our knowledge and understanding of the central nervous system in deep hypothermic conditions, thus demonstrating the protective effects of cerebral and spinal cord cooling in patients with injuries as well as the ability of the brain to survive extended periods of total circulatory arrest at extremely low temperatures. It was there that isolated brain preparation and transplantation were first accomplished. These and other unique, surgically constructed brain models opened new fields of exploration in neurochemical, neurophysiological, rheological, immunological, and cognitive features of the brain in normothermic and various hypothermic states. During the laboratory's most productive years (1961-1976), there were 10 surgeons actively involved in scientific investigations who later became chairmen or chiefs of Departments or Divisions of Neurosurgery and another four who became professors of neurosurgery or other surgical specialties. PMID- 15540935 TI - Interoperative angiography. PMID- 15540936 TI - Pavlov and Cushing. PMID- 15540937 TI - Enzyme-activated, hypoxia-selective DNA damage by 3-amino-2 quinoxalinecarbonitrile 1,4-di-N-oxide. AB - The compound 3-amino-2-quinoxalinecarbonitrile 1,4-dioxide (4) displays potent hypoxia-selective cytotoxicity in cell culture. This compound is structurally similar to the known hypoxia-selective DNA-damaging agent tirapazamine (1, TPZ), but the ability of 4 to cause DNA damage under low-oxygen conditions has not previously been characterized. The results presented here provide the first evidence that 4 causes reductively activated DNA damage under hypoxic conditions. The findings indicate that one-electron reduction of 4 by NADPH:cytochrome P450 reductase yields an oxygen-sensitive intermediate (5). This activated intermediate is rapidly destroyed by reaction with O2 under aerobic conditions, but goes forward to cause DNA damage under low-oxygen conditions. Analysis of the DNA damage indicates that reductive activation of 4 leads to production of a highly reactive, freely diffusible oxidizing radical that causes sequence independent cleavage of the deoxyribose backbone and oxidative damage to the heterocyclic bases in duplex DNA. On the basis of the experiments reported here, the chemical nature of the DNA damage caused by redox-activated 4 is analogous to that reported previously for TPZ. PMID- 15540938 TI - 5'-(2-phosphoryl-1,4-dioxobutane) as a product of 5'-oxidation of deoxyribose in DNA: elimination as trans-1,4-dioxo-2-butene and approaches to analysis. AB - Oxidation of deoxyribose in DNA leads to the formation of a spectrum of electrophilic products unique to each position in the sugar. For example, chemical reactions following abstraction of the C5'-hydrogen atom partition to form either a nucleoside 5'-aldehyde residue attached to the 5'-end of the DNA strand or a 5'-formyl phosphate residue attached to the 3'-end of the DNA strand that is accompanied by a four-carbon fragment on the 5'-end. We now present two approaches that both identify the latter fragment as 5'-(2-phosphoryl-1,4 dioxobutane) and provide a means to quantify the formation of this residue by different oxidizing agents. The first approach involves oxidation of DNA followed by reaction with O-benzylhydroxylamine to form stable dioxime derivatives of the putative 5'-(2-phosphoryl-1,4-dioxobutane) residues. The beta-elimination product of this dioxime proved to be the expected trans-1,4-dioxo-2-butene, as judged by gas chromatographic and mass spectrometric (GC/MS) comparison to authentic dioximes of cis- and trans-1,4-dioxo-2-butene, which revealed a unique pattern of three signals for each isomer, and by X-ray crystallography. Using a benzylhydroxylamine dioxime derivative of [2H4]-labeled cis-1,4-dioxo-2-butene as an internal standard, the dose-response for the formation of 5'-(2-phosphoryl-1,4 dioxobutane) was determined to be linear for gamma-radiation, with approximately 6 lesions per 10(6) nt per Gy, and nonlinear for Fe2+-EDTA. A comparison of 5'-(2 phosphoryl-1,4-dioxobutane) formation to total deoxyribose oxidation suggests that gamma-radiation produces approximately 0.04 lesions per deoxyribose oxidation event. As a positive control for 5'-oxidation of deoxyribose, the enediyne calicheamicin was observed to produce 5'-(2-phosphoryl-1,4-dioxobutane) at the rate of approximately 9 lesions per 10(6) nt per microM. A second approach to identifying and quantifying the sugar residue involved derivatization with hydrazine and beta-elimination to form pyridazine followed by quantification of the pyridazine by GC/MS. Using this approach, it was observed that the enediyne, neocarzinostatin, produced a linear dose-response for pyridazine formation, as expected given the ability of this oxidant to cause 1'-, 4'-, and 5'-oxidation of deoxyribose in DNA. The antitumor antibiotic, bleomycin, on the other hand, produced pyridazine at a 10-fold lower rate, which is consistent with 4' chemistry as the predominant mode of deoxyribose oxidation by this agent. These results provide novel insights into the chemistry of deoxyribose oxidation in DNA and two approaches to quantifying the 5'-(2-phosphoryl-1,4-dioxobutane) precursor of trans-1,4-dioxo-2-butene, an electrophile known to react with nucleobases to form novel DNA adducts. PMID- 15540939 TI - Isolation of 41a-homoyessotoxin and the identification of 9-methyl-41a homoyessotoxin and nor-ring A-yessotoxin from Protoceratium reticulatum. AB - 41a-homoyessotoxin (1), a new analogue of yessotoxin (7), was isolated from extracts of Protoceratium reticulatum grown in culture. In addition, 9-methyl-41a homoyessotoxin (2) and a nor-ring A-yessotoxin (3) were identified as the major components of a mixed fraction. The structural information was initially obtained from LC-UV and LC-MS3 chromatograms, with subsequent definitive structure determination by NMR, LC-MS/MS, and high-resolution MS. Full 1H and indirectly detected 13C NMR assignments for all but two carbon atoms were obtained from ca. 100 microg of 1. Full 1H NMR assignments for 2 and 3 and identification of three new heptanor-41-oxo-analogues of 3 (4-6) during LC-MS3 analysis of a fraction containing 1-3 and 7 are also reported. PMID- 15540940 TI - A novel pectenotoxin, PTX-12, in Dinophysis spp. and shellfish from Norway. AB - Two novel pectenotoxins (PTXs) were detected by LC-MS in solid phase extracts of net hauls taken at Flodevigen, Norway, in June 2002 that were dominated by Dinophysis acuminata and Dinophysis norvegica. The new compounds were isolated as minor components from a large collection of a Dinophysis acuta-dominated bloom obtained from Skjer, Sognefjorden, Norway, in October 2002. LC-MS and NMR analyses revealed that the new components, 36S-PTX-12 and 36R-PTX-12, occurred as a pair of equilibrating diastereoisomers differing from PTX-2 in that they contained an exocylic olefinic methylene rather than a methyl group at C-38. Analyses of shellfish extracts revealed that PTX-12 accumulated in Norwegian blue mussels (Mytilus edulis) and cockles (Cerastoderma edule), along with PTX-12 seco acids occurring as a complex mixture of diastereoisomers. LC-MS analysis of algal cells picked from the net haul from Flodevigen revealed that PTX-12 predominated in D. acuta and D. norvegica, whereas PTX-2 was the predominant pectenotoxin in D. acuminata. Preliminary observations indicate that the relative contents of PTX 2 and PTX-12 vary between sites and years in Norway, even within a single species of Dinophysis. Our data also suggest that heterotrophic dinoflagellates may accumulate toxins from their prey. PMID- 15540941 TI - Experimental and theoretical studies on the pharmacodynamics of cisplatin in jurkat cells. AB - For Jurkat cells in culture exposed to cisplatin (1), we measured the number of platinum adducts on DNA and showed that it is proportional to the AUC, the area under the concentration vs time curve, for cisplatin. The number of platinum-DNA adducts is measured immediately following exposure to drug. The AUC is calculated either as the product of the initial cisplatin concentration and the exposure time or as the integral under the concentration vs time curve for the unreacted dichloro species, which decreases exponentially. We also show that the number of adducts correlates with decreases in respiration, with the amount of DNA fragmentation, and with cell viability, all measured 24 h after exposure to the drug. To study the reactions of cisplatin at concentrations approaching clinical relevance (65 microM), we use two-dimensional [1H15N]HSQC NMR and the 15N-labeled form of the drug, cis-Pt(15NH3)2Cl2, 1. In the absence of cells, 1 reacts with components of the growth medium and also transforms slowly (k(h) = 0.205 h-1 at 37 degrees C) into the chloro-aquo species, cis-[Pt(15NH3)2Cl(H2O)]+ (2), which at the pH of the medium (pH 7.15), is mainly in the deprotonated chloro-hydroxy form, cis-Pt(15NH3)2Cl(OH) (4). The concentration of 2 (4), as measured by HSQC NMR, decreases due to reaction with components of the medium. In the presence of 5 million or more cells, the concentration of 1 decreases with time, but the NMR signal for 2 (4) is not seen because it is rapidly removed from solution by the cells, keeping its concentration very low. These experiments confirm that the species preferentially removed from the medium by cells is 2 (4) and not 1. Our findings are discussed in the context of a kinetic model for platination of nuclear DNA by cisplatin, which includes aquation of cisplatin outside the cell, passage of 2 (4) through the cell membrane, reaction of reactive platinum species (RPS) in the cytosol with thiols, formation of adducts between RPS and accessible sites on genomic DNA, and removal of platinum from DNA by repair. Some of the rate constants involved are measured, but others can only be estimated. Calculations with this model show that little of the platinum reacts with intracellular thiols before reaching the nuclear DNA, indicating that binding to thiols is not important in cisplatin resistance. The model also predicts the circumstances under which the amount of platination of nuclear DNA is proportional to AUC. PMID- 15540942 TI - Oxidative DNA damage induced by benz[a]anthracene dihydrodiols in the presence of dihydrodiol dehydrogenase. AB - Tobacco smoke and polluted air are risk factors for lung cancer and contain many kinds of polycyclic aromatic hydrocarbons (PAHs) including benzo[a]pyrene (B[a]P) and benz[a]anthracene (BA). BA, as well as B[a]P, is assessed as probably carcinogenic to humans (IARC group 2A). BA is metabolized to several dihydrodiols. Dihydrodiol dehydrogenase (DD), a member of the aldo-keto reductase superfamily, catalyzes NAD(P)+-linked oxidation of dihydrodiols of aromatic hydrocarbons to corresponding catechols. To clarify the role of DD on PAH carcinogenesis, we examined oxidative DNA damage induced by trans-dihydrodiols of BA and B[a]P treated with DD using 32P-5'-end-labeled DNA fragments obtained from the human p53 tumor suppressor gene. In addition, we investigated the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), an indicator of oxidative DNA damage, in calf thymus DNA by using HPLC with an electrochemical detector. DD catalyzed BA-1,2-dihydrodiol caused Cu(II)-mediated DNA damage including 8-oxodG formation in the presence of NAD+. BA-1,2-dihydrodiol induced a Fpg sensitive and piperidine labile G lesion at the 5'-ACG-3' sequence complementary to codon 273 of the human p53 tumor suppressor gene, which is known as a hotspot. DNA damage was inhibited by catalase and bathocuproine, suggesting the involvement of H2O2 and Cu(I). The observation of NADH production by UV-visible spectroscopy suggested that DD catalyzed BA-1,2-dihydrodiol most efficiently to the corresponding catechol among the PAH-dihydrodiols tested. A time-of-flight mass spectroscopic study showed that the catechol form of BA-1,2-dihydrodiol formed after DD treatment. In conclusion, BA-1,2-dihydrodiol can induce DNA damage more efficiently than B[a]P-7,8-dihydrodiol and other BA-dihydrodiols in the presence of DD. The reaction mechanism on oxidative DNA damage may be explained by theoretical calculations with an enthalpy change of dihydrodiols and oxidation potential of their catechol forms. DD may play an important role in BA carcinogenesis via oxidative DNA damage. PMID- 15540943 TI - Co(II) and Cd(II) substitute for Zn(II) in the zinc finger derived from the DNA repair protein XPA, demonstrating a variety of potential mechanisms of toxicity. AB - XPA is one of the key members of the protein complex of the nucleotide excision repair (NER) pathway of DNA repair. The CCCC zinc finger domain of XPA is involved in the interactions with other NER proteins. To study the possible molecular mechanisms of XPA inhibition, we previously investigated Zn(II) and Ni(II) interactions with the synthetic 37 amino acid peptide (XPAzf), AcDYVICEECGKEFMDSYLMNHFDLPTCDNCRDADDKHKam, representing the XPA zinc finger sequence (Bal, W., Schwerdtle, T., and Hartwig, A. (2003) Mechanism of nickel assault on the zinc finger of DNA repair protein XPA. Chem. Res. Toxicol. 16, 242 248). In this work, we extended these studies on other carcinogenic metal ions, Co(II) and Cd(II). The binding constants and complex geometries were determined using UV-vis and CD spectroscopies, and oxidative damage to XPAzf was studied with HPLC. The conditional binding constants determined for Co(II) and Cd(II) in 50 mM phosphate buffer, pH 7.4, are 10(7.4)+/-(0.4) and 10(12.8)+/-(0.5), respectively, yielding binding constant ratios Zn(II)/Co(II) of 100 and Zn(II)/ Cd(II) of 0.001, which are the lowest values reported for zinc fingers so far. The Co(II) ion forms a tetrahedral complex with the sulfurs of XPAzf, which is isostructural with the native zinc finger. The Cd(II) complex is somewhat less structured. The oxidation of Zn(II)-saturated XPAzf by H2O2 is accelerated in the presence of Co(II), but the concentration profile of this effect indicates the formation of an active Co(II) complex external to the metal-sulfur center. The Cd(II)-saturated XPAzf is very resistant to oxidation by H2O2. Overall, our results indicate that XPAzf can undergo Co(II) and Cd(II) assault under specific conditions. PMID- 15540945 TI - Uranium(VI) solubility and speciation in simulated elemental human biological fluids. AB - The complete understanding of the human body response to uranium contamination exposure is vital to the development of exposure analysis and subsequent treatments for overexposure. Thermodynamic modeling has traditionally been used to study environmental metal contaminant migration (especially uranium and other radionuclides), allowing examination of chemical processes difficult to study experimentally. However, such techniques are rarely used in the study of metal toxicology. Chemical thermodynamics has a unique and valuable role in developing models to explain metal metabolism and toxicology. Previous computational models of beryllium in simulated biological fluids have been shown to be useful in predicting metal behavior in the human body. However, previous studies utilizing chemical thermodynamics in understanding uranium chemistry in body fluids are limited. Here, a chemical thermodynamic speciation code has been used to model and understand the chemistry of uranium in simulated human biological fluids such as intracellular, interstitial, and plasma fluids, saliva, sweat, urine, bile, gastric juice, pancreatic fluid, and a number of airway surface fluids from patients with acute lung conditions. The results show predicted uranium solubility, and speciation varies markedly between each biological fluid due to differences in fluid composition, ionic strength, and pH. The formation of uranium hydroxide, phosphate (sodium/potassium autunite), and calcium uranate was observed in most of the fluids. The results of this work, supported by experimental validation, can aid in understanding the metabolism and toxic effects of uranium with potential applications to biological monitoring as well as chelation treatment of uranium body burden. PMID- 15540944 TI - Inhibition of Hsp72-mediated protein refolding by 4-hydroxy-2-nonenal. AB - A proteomic approach was applied to liver cytosol from rats fed a diet consisting of high fat and ethanol to identify 4-hydroxy-2-nonenal (4-HNE)-modified proteins in vivo. Cytosolic Hsp72, the inducible variant of the Hsp70 heat shock protein family, was consistently among the proteins modified by 4-HNE. Despite 1.3-fold induction of Hsp72 in the livers of ethanol-fed animals, no increase in Hsp70 mediated luciferase refolding in isolated heptocytes was observed, suggesting inhibition of this process by 4-HNE. A 50% and 75% reduction in luciferase refolding efficiency was observed in rabbit reticulocyte lysate (RRL) supplemented with recombinant Hsp72 which had been modified in vitro with 10 and 100 microM 4-HNE, respectively. This observation was accompanied by a 25% and 50% decrease in substrate binding by the chaperone following the same treatment; however, no effect on complex formation between Hsp72 and its co-chaperone Hsp40 was observed. Trypsin digest and mass spectral analysis of Hsp72 treated with 10 and 100 microM 4-HNE consistently identified adduct formation at Cys267 in the ATPase domain of the chaperone. The role of this residue in the observed inhibition was demonstrated through the use of DnaK, a bacterial Hsp70 variant lacking Cys267. DnaK was resistant to 4-HNE inactivation. Additionally, Hsp72 was resistant to inactivation by the thiol-unreactive aldehyde malondialdehyde (MDA), further supporting a role for Cys in Hsp72 inhibition by 4-HNE. Finally, the affinity of Hsp72 for ATP was decreased 32% and 72% following treatment of the chaperone with 10 and 100 microM 4-HNE, respectively. In a model of chronic alcoholic liver injury, induction of Hsp72 was not accompanied by an increase in protein refolding ability. This is likely the result of 4-HNE modification of the Hsp72 ATPase domain. PMID- 15540946 TI - Novel identification of a sulfur-centered, radical-derived 5,5-dimethyl-1 pyrroline N-oxide nitrone adduct formed from the oxidation of DTT by LC/ELISA, LC/electrospray ionization-MS, and LC/tandem MS. AB - The detection of highly reactive free radicals generated in biological systems by an ESR spin-trapping technique is always difficult and limited due to the short lifetimes of ESR active spin-trapping radical adducts and poor structural information provided by ESR spectra. In this investigation, we have for the first time employed anti-5,5-dimethyl-1-pyrroline N-oxide (DMPO) polyclonal antiserum that specifically recognizes stable, ESR silent end products of DMPO radical adducts and combined HPLC with ELISA, electrospray ionization mass spectrometry (ESI-MS), and tandem mass spectrometry (MS/MS) to separate and characterize DMPO nitrone adducts derived from free radical metabolites. When mircoperoxidase-11 (MP-11) reacted with DTT in the presence of DMPO with or without H2O2, we detected radical-derived DMPO nitrone adducts by ELISA. Similar results were obtained when MP-11 was replaced by hemin. To identify the DMPO nitrone adducts formed in both reaction systems, LC separation was carried out, and the fractions eluted from the LC column were collected and analyzed by ELISA. In both reaction mixtures, we found that only one peak with the same retention time showed a strong positive ELISA signal, suggesting that this peak was from radical-derived DMPO nitrone adducts and that both systems produced the same free radical metabolites. Using online LC/ESI-MS, LC/MS/MS, and (1)H NMR, we demonstrated that the DMPO nitrone adducts formed are from the DMPO adducts of the sulfur-centered radical of DTT. The successful application of LC/ELISA, LC/MS, and LC/MS/MS in this study makes it possible to separate and identify the stable DMPO nitrone adducts derived from free radical metabolites generated in biological systems. PMID- 15540947 TI - Further development of 32P-postlabeling for the detection of alkylphosphotriesters: evidence for the long-term nonrandom persistence of ethyl phosphotriester adducts in vivo. AB - DNA phosphate oxygens are sites for alkylation leading to phosphotriester adducts (PTEs). PTEs are reported to be both abundant and persistent and so may serve as long-term markers of genotoxicity. Previously, we reported a 32P-postlabeling assay for the specific detection of PTEs plus identification of nucleosides located 5' to PTEs. Using this, we demonstrated the nonrandom nature of ethyl PTEs (Et-PTEs) in vivo, these results being suggestive of either the nonrandom formation of Et-PTEs in vivo or sequence specific Et-PTE repair. Presently, we report the further development and validation of the 32P-postlabeling assay, to permit the more straightforward determination of nucleosides 5' to PTEs and, using this, have investigated the long-term persistence of PTEs in vivo. Analysis of liver DNA of mice treated in vivo with N-nitrosodiethylamine reveals an initial decline in the level of Et-PTEs (t1/2<24 h) as well as their nonrandom persistence for the duration of the time course, with approximately 37 and approximately 15% of the initial Et-PTEs remaining 4 and 56 days after treatment, respectively. From this, we conclude that Et-PTEs are suitable as long-term markers of genotoxic exposure and that putative PTE repair is not responsible for their nonrandom manifestation. However, the possibility of active repair contributing to the initial decline of Et-PTEs is considered. PMID- 15540948 TI - Mass spectrometric identification of 4-hydroxy-2,5-dioxo-imidazolidine-4 carboxylic acid during oxidation of 8-oxoguanosine by peroxynitrite and KHSO5/CoCl2. AB - Several products were previously identified from the reaction of 8-oxodG with bolus additions of peroxynitrite. Those experiments involved very high instantaneous concentrations of peroxynitrite and raised the possibility that the product spectrum may have differed from what might occur in vivo where much lower -and steadier--peroxynitrite levels would be expected. We consequently examined this issue by treating 2',3',5'-tri-O-acetyl-8-oxoGuo with infused authentic peroxynitrite and 3-morpholinosydnonimine and have found that under these conditions an additional product, not found under bolus addition conditions, was observed. Using tandem mass spectrometry, isotope labeling, and synthesis of a structural analogue, this compound was identified as 4-hydroxy-2,5-dioxo imidazolidine-4-carboxylic acid (HICA). The mechanism of HICA formation is rationalized by water attack at C5 of a quininoid intermediate. PMID- 15540949 TI - Spiroiminodihydantoin and guanidinohydantoin are the dominant products of 8 oxoguanosine oxidation at low fluxes of peroxynitrite: mechanistic studies with 18O. AB - Peroxynitrite-mediated oxidation of 8-oxoguanosine results in the formation of two product classes distinguished by the source of their incorporated oxygen atoms. The first product class consists of dehydroguanidinohydantoin (DGh), N nitro-dehydroguanidinohydantoin (NO2-DGh), and 2,4,6-trioxo[1,3,5]triazinane-1 carboxamidine (CAC) with peroxynitrite as the exogenous O atom source, and the second includes spiroiminodihydantoin (Sp), guanidinohydantoin (Gh), and 4 hydroxy-2,5-dioxo-imidazolidine-4-carboxylic acid (HICA), with water serving as the exogenous O atom source. The first product class forms exclusively at high peroxynitrite fluxes, while the second forms exclusively at limiting peroxynitrite fluxes. At intermediate peroxynitrite fluxes, both sets of products are formed. At high fluxes, DGh was the major reaction product, and after several of the peroxynitrite-derived radicals were eliminated as the exogenous O atom source, the peroxynitrite anion emerged as the most likely candidate. On the other hand, at lower fluxes, either Gh or Sp was the major product, depending on the pH of the reaction mixture. At low and high pH, respectively, Gh and Sp were the major products, and the plot of pH vs ratio of Sp/(Sp+Gh) had an inflection at pH 5.8. Interestingly, the pH dependence for oxidation of 8-oxoGuo with CoCl2 and KHSO5 was identical to that for oxidation by peroxynitrite, indicating that the phenomenon arises due to characteristics of an 8-oxoGuo-derived rather than an oxidant-derived intermediate, since these two systems generate different reactive species. On the basis of these findings, a model in which 8-oxoGuo is oxidized to the bisimine intermediate, 1 is proposed. At high peroxynitrite fluxes, the reaction of 1 with ONOO- predominates over the reaction with H2O, leading exclusively to DGh, NO2-DGh, and CAC, while at limiting peroxynitrite concentrations, the reaction with H2O dominates, and Gh and Sp are formed exclusively. At intermediate peroxynitrite fluxes, the relative kinetics of the reaction between 1 and ONOO- or H2O are such that both product classes are formed. To explain the pH-dependent Gh and Sp yields, we propose that 5 has a pKa approximately 5.8 and that the differential reactivity of the protonated and deprotonated form of 5 leads to its partitioning into Gh and Sp, respectively. PMID- 15540950 TI - Identification of 14 quercetin phase II mono- and mixed conjugates and their formation by rat and human phase II in vitro model systems. AB - In this study, the HPLC, UV-vis, LC-MS, and 1H NMR characteristics of 14 different phase II mono- and mixed conjugates of quercetin were determined, providing a useful tool in the identification of quercetin phase II metabolite patterns in various biological systems. Using these data, the phase II metabolism of quercetin by different rat and human liver and intestine in vitro models, including cell lines, S9 samples, and hepatocytes, was investigated. A comparison of quercetin phase II metabolism between rat and human liver and intestinal cell lines, S9, and hepatocytes showed considerable variation in the nature and ratios of quercetin conjugate formation. It could be established that the intestine contributes significantly to the phase II metabolism of quercetin, especially to its sulfation, that organ-dependent phase II metabolism in rat and man differ significantly, and that human interindividual variation is higher for quercetin sulfation than for glucuronidation or methylation. Furthermore, quercetin conjugation by different in vitro models from corresponding origins may differ significantly. The identification of the various mono- and mixed quercetin phase II conjugates revealed significant differences in phase II conjugation by a variety of in vitro models and led to the conclusion that none of the in vitro models converted quercetin to a phase II metabolite mixture similar to the in vivo plasma metabolite pattern of quercetin. Altogether, the identification of a wide range of phase II metabolites of quercetin as presented in this study allows the determination of quercetin phase II biotransformation patterns and opens the way for a better-funded assessment of the biological activity of quercetin in a variety of biological systems. PMID- 15540951 TI - Determinants of selectivity in alkylation of nucleosides and DNA by secondary diazonium ions: evidence for, and consequences of, a preassociation mechanism. AB - Reactions have been carried out in which 1,3-diisopropyltriazene or N-isopropyl-N (1-hydroxyethyl)nitrosamine has been decomposed in neutral, buffered aqueous media in the presence of (15N2)2'-deoxyguanosine and (15N6)2'-deoxyadenosine. The products of covalent attachment of the isopropyl cation, derived from the isopropyl diazonium ion, to the heteroatoms of the purines have been separated and quantified by HPLC/electrospray mass spectrometry by employing isotopically distinct synthetic standards. The results indicate that the two different precursors of the isopropyl cation result in the formation of different yields of products in the reactions at all of the heteroatoms of both purines, outside experimental error, except possibly in the case of the N3 position of dAdo. For the different alkylating agents, the ratios of yields at any two sites vary as well. This leads to the conclusion that isopropylation occurs by a preassociation mechanism in which the isopropyl cation intermediate reacts in the solvation shell in which it is generated from its precursors. The reaction of N-isopropyl-N (1-hydroxyethyl)nitrosamine results in alkylation of 2'-deoxyguanosine in preference to 2-deoxyadenosine, by a factor of 3-4. In this reaction, the yields for reaction at N1, N3, N6, and N7 of adenine vary over less than a factor of 2, whereas the yields for N2, N3, O6, and N7 of guanine vary over less than a factor of 4. The N1 atom of guanine is disfavored over the major product, the O6 adduct, by a factor of <8. The reaction of 1,3-diisopropyltriazene shows a similar preference for alkylation of 2-deoxyguanosine, with a similar range of product distribution in the reactions at adenine heteroatoms and a somewhat larger range of distribution at guanine heteroatoms. In particular, the yield of 1 isopropylguanosine is 50-fold lower than that of O6-isopropylguanosine. The comparable yields of products of reaction at the "hard" and "soft" sites of the purines lead to the conclusion that nucleophilicity is unimportant in site selectivity of alkylation by the isopropyl cation. The noteworthy selectivities, above, are rationalized by: differences in the association constants of the precursors of the cations with the two nucleosides; steric, statistical, and electrostatic effects that favor reaction of the O6 atom of guanine; and larger steric and/or desolvation requirements for association of the 1,3 diisopropyltriazenium cation with the N1 atom of guanine. The reaction of N isopropyl-N-(1-hydroxyethyl)nitrosamine with double-stranded DNA has been similarly analyzed. The product distribution is remarkably similar in profile to that observed for the nucleosides in solution. In particular, exocyclic amino groups are competitive with the more traditional sites of diazonium ion-mediated alkylation. A comparison to earlier literature data on alkylation by methyl- and ethyl-diazonium ions illustrates some fundamental differences between the reaction of the diazonium ions and the isopropyl cation derived from the isopropyl diazonium ion. PMID- 15540952 TI - Rotenone, deguelin, their metabolites, and the rat model of Parkinson's disease. AB - Rotenone and deguelin are the major active ingredients and principal components of cuberesin from Lonchocarpus utilis used as a botanical insecticide and piscicide. They are also potent complex I (NADH:ubiquinone oxidoreductase) inhibitors. Rotenone was known earlier, and deguelin is shown here to induce a Parkinson's disease (PD)-like syndrome after subcutaneous treatment of rats by osmotic minipump. Rotenone at 3 mg/kg/day or deguelin at 6 but not 3 mg/kg/day induces degeneration of the nigrostriatal dopaminergic pathway, as shown by reduced tyrosine hydroxylase immunoreactivity with treatments for 5 or 6 days. The neuropathological lesions are associated with a brain level of parent rotenoid of 0.4-1.3 ppm but not with the much smaller brain level of 12abeta hydroxyrotenoids or other metabolites analyzed by HPLC and LC/MS. We previously established that the hydroxylated metabolites and derivatives of rotenone and deguelin are all less active (i.e., detoxified) as complex I inhibitors relative to the parent rotenoids. The PD-like syndrome induced in rats by rotenone and deguelin is therefore due to the parent compounds rather than metabolites. Deguelin is about half as active as rotenone in inducing the PD-like syndrome in rats and in acute ip LD50 in mice. Rotenone and deguelin are metabolized by human recombinant 3A4 and 2C19 but not five other P450 enzymes. 2C19 is more selective than 3A4 in forming the 12abeta-hydroxyrotenoids. Identified sites of metabolic attack individually or in combination are as follows: 12abeta hydroxylation and 2 O-demethylation of both compounds, oxidation of the rotenone isopropenyl substituent to mono and diol derivatives, and probable oxidation of the deguelin dimethylchromene double bond. These toxicological features must be considered in using rotenone, deguelin, and their analogues as pesticides, candidate radioimaging and cancer chemopreventive agents, and models of PD. PMID- 15540956 TI - Cardiovascular safety of rofecoxib (Vioxx): lessons learned and unanswered questions: we need processes in place to follow up suspicions about serious adverse events. PMID- 15540957 TI - Atkins and the new diet revolution: is it really time for regimen change? AB - Weight loss occurs in the short term, but not enough is known to recommend long term use. PMID- 15540958 TI - Improved evidence-based management of acute musculoskeletal pain: guidelines from the National Health and Medical Research Council are now available. PMID- 15540959 TI - The anti-TNF revolution in ankylosing spondylitis: patients with severe disease now have access to promising new drugs. PMID- 15540961 TI - Towards health equity through an adult health check for Aboriginal and Torres Strait Islander people: an important Australian initiative that sets an international precedent. PMID- 15540963 TI - The energy of slaves. PMID- 15540964 TI - Effectiveness and side effects of thiazolidinediones for type 2 diabetes: real life experience from a tertiary hospital. AB - OBJECTIVE: To assess effectiveness and side effects of thiazolidinediones (TZDs) as adjunctive therapy in suboptimally controlled patients with type 2 diabetes. DESIGN AND SETTING: Review of a prospectively recorded database at the Royal Melbourne Hospital diabetes clinic. PARTICIPANTS: 203 patients with type 2 diabetes who received pioglitazone or rosiglitazone between 1 May 2000 and 31 October 2002. OUTCOME MEASURES: Response in glycohaemoglobin (HbA(1c)) level, lipid profile changes and side effects, including hypoglycaemia, weight gain, oedema and precipitation of cardiac failure. RESULTS: Both pioglitazone and rosiglitazone improved glycaemic control, with a reduction in the HbA(1c) level of 1.02% (range, 0.85%-1.19%) and 0.96% (range, 0.81%-1.11%), respectively, in the first 6 months of therapy. Rosiglitazone was associated with a 0.45 mmol (range, 0.31-0.59 mmol) increase in cholesterol level and 0.99 mmol (range, 0.60 1.38 mmol) increase in triglyceride level, while pioglitazone was associated with insignificant declines in cholesterol and triglyceride levels. There was reduced requirement for insulin, but not for oral hypoglycaemic agent (OHA), in most patients who used these agents. Pioglitazone and rosiglitazone were associated with increased rates of hypoglycaemia (17% and 11% of patients, respectively), significant weight gain (48% and 58%) and oedema (33% and 21%). There were four cases of acute left ventricular failure and two cases of reversible liver dysfunction in patients treated with TZDs. CONCLUSIONS: Adding pioglitazone or rosiglitazone therapy to OHA or insulin in patients with type 2 diabetes significantly improved glycaemic control. However, the use of these drugs in routine clinical practice was associated with more frequent adverse events than previously reported in clinical trials. PMID- 15540965 TI - How does mental health status relate to accessibility and remoteness? AB - OBJECTIVE: To determine whether mental illness is associated with accessibility and remoteness. DESIGN: A cross-sectional, population-based, computer-assisted telephone interview survey, stratified by Accessibility and Remoteness Index of Australia (ARIA) categories. SETTING: Secondary analysis of data collected from 2545 South Australian adults in October and November 2000. OUTCOME MEASURES: Psychological distress and depression as determined by the Kessler 10 Psychological Distress Scale, the SF-12 measure of health status, and self reported mental illness diagnosed by a doctor in the previous 12 months. RESULTS: Overall, mental illness prevalence estimates were similar using the three measures of psychological distress (10.5%), clinical depression (12.9%) and self reported mental health problem (12.7%). For each measure, there was no statistically significant variation in prevalence across ARIA categories, except for a lower than expected prevalence of depression (7.7%) in the "accessible" category. There was no trend suggesting higher levels of mental illness among residents of rural and remote regions. CONCLUSIONS: The prevalence rates of psychological distress, depression and self-reported mental illness are high. However, we found no evidence that the prevalence of these conditions varies substantially across ARIA categories in South Australia. This finding may challenge existing stereotypes about higher levels of mental illness outside metropolitan Australia. PMID- 15540966 TI - Diabetes, psychotic disorders and antipsychotic therapy: a consensus statement. AB - Psychotic illness and its treatment are associated with an increased rate of diabetes and worsening blood sugar control, Australia. The newer, second generation antipsychotic agents are more likely to produce this effect than the first-generation agents, but both contribute to the problem. The effect is usually related to insulin resistance through weight gain, but other mechanisms may exist. Diabetic ketoacidosis is rare. Management of psychosis takes priority over concerns about the potential metabolic sequelae of treatment, but the prevalence of the latter requires that all patients taking antipsychotic agents be actively screened and treated. Patients treated with antipsychotic agents need baseline and regular checks, including weight, blood glucose and lipid levels and blood pressure. Management of psychosis with its attendant medical problems requires a multidisciplinary approach, with primary health practitioners playing a central role. Mortality and medical morbidity is higher in those with psychosis than expected; preventive measures, combined with early detection and treatment of hyperglycaemia and other metabolic problems, is a key public health issue. PMID- 15540967 TI - The dearth of new antibiotic development: why we should be worried and what we can do about it. AB - The emergence and spread of multidrug-resistant pathogens has increased substantially over the past 20 years. Over the same period, the development of new antibiotics has decreased alarmingly, with many pharmaceutical companies pulling out of antibiotic research in favour of developing "lifestyle" drugs. Reasons given for withdrawing from antibiotic development include poor "net present value" status of antibiotics, changes in regulations requiring larger drug trials and prolonged post-marketing surveillance, clinical preference for narrow-spectrum rather than broad-spectrum agents, and high new-drug purchase costs. Major improvements in infection control in Australia are needed to prevent further spread of resistant clones, buying some time to develop urgently needed new antibiotic agents. Perpetuating a culture of "pharma bashing" will simply lead to more pharmaceutical companies withdrawing from the market. A change in the health and research culture is needed to improve cooperation between public, academic and private sectors. PMID- 15540969 TI - Does probiotic milk prevent infections in children attending daycare centres? PMID- 15540968 TI - Partial oesophageal perforation associated with cold carbonated beverage ingestion. AB - We present a rare case of spontaneous intramural oesophageal perforation after the rapid ingestion of a cold carbonated beverage. A previously well patient presented with sudden onset of severe retrosternal pain associated with pain on swallowing. A contrast computed tomography scan and gastroscopy confirmed the diagnosis. With the widespread popular practice of drinking cold carbonated beverages, especially during the summer season, clinicians should be aware of this possible serious complication. PMID- 15540970 TI - Generalising the results of trials to clinical practice. PMID- 15540971 TI - 2. Acute infectious diarrhoea and dehydration in children. AB - Gastroenteritis in children is still a common reason for consulting a general practitioner and for hospital admission. Rotavirus is the most common cause of gastroenteritis in children and accounts for half of all hospital admissions for severe acute infectious diarrhoea. Most children with gastroenteritis do not develop dehydration and can be treated at home. Children with mild to moderate dehydration should be treated with low osmolarity oral rehydration solutions, and those with severe dehydration or shock need to be admitted for administration of intravenous fluids. Lactose-free feeds should not be routinely used after acute gastroenteritis, but there is some evidence that a lactose-free diet may reduce the duration of diarrhoea. Antimotility drugs are rarely indicated in children with gastroenteritis, as the potential risks outweigh the benefits. The development of a rotavirus vaccine would provide huge public health benefits and cost savings. Other preventive strategies include educating people about personal and food hygiene and encouraging breastfeeding. PMID- 15540972 TI - Prescribing of amino acid infant formula. PMID- 15540973 TI - A2 milk is allergenic. PMID- 15540974 TI - Rectal perforation from colonic irrigation administered by alternative practitioners. PMID- 15540975 TI - Critical shortage of injectable thiamine in Australia. PMID- 15540976 TI - To exercise or not to exercise in chronic fatigue syndrome? PMID- 15540977 TI - Pertussis vaccination for new parents? PMID- 15540978 TI - Three Australian whistleblowing sagas: lessons for internal and external regulation. PMID- 15540979 TI - Institutional racism in Australian healthcare: a plea for decency. PMID- 15540980 TI - Ethical and legal issues at the interface of complementary and conventional medicine. PMID- 15540981 TI - Timing of health assessments. PMID- 15540982 TI - UK health inequalities: the class system is alive and well. PMID- 15540983 TI - Should telemedicine in eye care be funded in Australia? PMID- 15540984 TI - Drugs, sport and the Olympics 2000-2004. PMID- 15540985 TI - Modulation of the p38 MAPK (mitogen-activated protein kinase) pathway through Bcr/Abl: implications in the cellular response to Ara-C. AB - The chimaeric protein Bcr/Abl, the hallmark of chronic myeloid leukaemia, has been connected with several signalling pathways, such as those involving protein kinase B/Akt, JNK (c-Jun N-terminal kinase) or ERKs (extracellular-signal regulated kinases) 1 and 2. However, no data about the p38 MAPK (mitogen activated protein kinase) have been reported. Here, we present evidence showing that Bcr/Abl is able to modulate this signalling pathway. Transient transfection experiments indicated that overexpression of Bcr/Abl in 293T cells is able to activate p38 MAPK or induce p73 stabilization, suggesting that c-Abl and Bcr/Abl share some biological substrates. Interestingly, the control exerted by Bcr/Abl on the p38 MAPK pathway was not only mediated by the tyrosine kinase activity of Bcr/Abl, as the use of STI571 demonstrated. In fact, Bcr alone was able to induce p38 MAPK activation specifically through MKK3 (MAP kinase kinase 3). Supporting these observations, chronic myeloid leukaemia-derived K562 cells or BaF 3 cells stably transfected with Bcr/Abl showed higher levels of phosphorylated p38 MAPK compared with Bcr/Abl-negative cells. While Bcr/Abl-negative cells activated p38 MAPK in response to Ara-C (1-beta-D-arabinofuranosylcytosine), Bcr/Abl-positive cells were unable to activate p38 MAPK, suggesting that the p38 MAPK pathway is not sensitive to Abl-dependent stimuli in Bcr/Abl-positive cells. Our results demonstrate that the involvement of Bcr/Abl in the p38 MAPK pathway is a key mechanism for explaining resistance to Ara-C, and could provide a clue for new therapeutic approaches based on the use of specific Abl inhibitors. PMID- 15540986 TI - Shedding and gamma-secretase-mediated intramembrane proteolysis of the mucin-type molecule CD43. AB - CD43 is a transmembrane molecule that contains a 123-aminoacids-long cytoplasmic tail and a highly O-glycosylated extracellular domain of mucin type. Endogenous CD43 expressed in COLO 205, K562 and Jurkat cells revealed a membrane-associated, 20 kDa CD43-specific cytoplasmic tail fragment (CD43-CTF) upon inhibition of gamma-secretase. This fragment was formed by an extracellular cleavage, as it was not accumulated after treating cells with 1,10-phenanthroline, a metalloprotease inhibitor. When CD43 was transfected into HEK-293 cells expressing dominant negative PS1 (presenilin-1), the CD43-CTF was accumulated, but not in cells with wild-type PS1. Owing to its accumulation in the presence of a non-functional PS variant, it may thus be a novel gamma-secretase substrate. This CTF is formed by an extracellular cleavage close to the membrane, is a fragment that can be concluded to be a substrate for gamma-secretase. However, the intracellular gamma secretase product has not been possible to detect, suggesting a quick processing of this product. During normal growth the CTF was not found without gamma secretase inhibition, but when the cells (COLO 205) were very confluent the fragment could be detected. The intracellular domain of CD43 has previously been shown to contain a functional nuclear localization signal, and has been suggested to be involved in gene activation. From this and the present results, a novel way to explain how mucin-type molecules may transduce intracellular signals can be proposed. PMID- 15540988 TI - Role of cytokines in cardiovascular diseases: a focus on endothelial responses to inflammation. AB - Complex cellular and inflammatory interactions are involved in the progress of vascular diseases. Endothelial cells, upon exposure to cytokines, undergo profound alterations of function that involve gene expression and de novo protein synthesis. The functional reprogramming of endothelial cells by cytokines is of importance especially in patients with chronic vascular inflammation. The intercellular network of dendritic cells, T-lymphocytes, macrophages and smooth muscle cells generates a variety of stimulatory cytokines [e.g. TNF-alpha (tumour necrosis factor-alpha), IL (interleukin)-1, IL-6 and IFN-gamma (interferon gamma)] and growth factors that promote the development of functional and structural vascular changes. High concentrations of proinflammatory cytokines increase oxidative stress, down-regulate eNOS (endothelial nitric oxide synthase) bioactivity and induce endothelial cell apoptosis. Chemoattractant cytokines [e.g. VEGF (vascular endothelial growth factor), TGF-beta1 (transforming growth factor-beta1) and IL-8] are important regulators of inflammation-induced angiogenesis and are directly modulated by nitric oxide. This review will focus on the vascular mechanisms orchestrated by cytokines and summarizes the current knowledge concerning the contribution of cytokines to cardiovascular diseases. PMID- 15540989 TI - Morphology and innervation pattern of the feline urogenital junction. AB - The feline urogenital junction is situated between the extratesticular rete and the spacious initial segments of the efferent ductules. The rete epithelium is cuboidal to low columnar. The rete cells forming the junction rest on a wavy basal lamina, display deep mutual invaginations, possess central nuclei with several infoldings and form a distinct border with the columnar epithelial cells of the initial segments of the ductuli efferentes. The epithelium of the initial segments is composed of ciliated cells and non-ciliated principal cells. The latter are the dominating type and characterized by an apical brush-border and a supranuclear endocytotic apparatus. The stroma of the extratesticular rete contains an abundance of collagen whereas contractile cells are here generally absent. In contrast, the initial segments of the efferent ductules are surrounded by elastic fibres and a layer of contractile cells. All nerves for the feline urogenital junction come from the nervus spermaticus superior. In the epididymal head, small nerve bundles deviate into the septa between the ductules. Single fibres establish a dense network within the muscular coat of the ductuli. At the transition to the extratesticular rete, this network ends abruptly. Nerve fibres in the confines of the rete are associated with blood vessels or proceed to the testicular interior, but establish no relationships with the rete epithelium or the myofibroblasts of the mediastinum. The nervous network in the walls of the efferent ductules and their initial segments is not only composed of sympathetic but also parasympathetic, non-myelinated fibres. Particularly noteworthy is the abundance of calcitonin gene-related peptide (CGRP)- and substance P (SP) containing axons around the initial segments. Both neuroproteins are consistent markers for sensory neurones. Taken together, it can be assumed that the entry of seminal fluid and spermatozoa into the efferent ductules is controlled by a regulatory nervous chain provided with afferent and efferent components. PMID- 15540987 TI - Lung Kruppel-like factor (LKLF) is a transcriptional activator of the cytosolic phospholipase A2 alpha promoter. AB - Increased expression of cPLA2 (cytosolic phospholipase A2) has been shown to be the cause of tumorigenesis of NSCLC (non-small-cell lung cancer). Our laboratory has previously demonstrated that oncogenic forms of Ras increase transcription of cPLA2 in normal lung epithelial cells and NSCLC lines through activation of the ERK (extracellular-signal-regulated kinase) and JNK (c-Jun N-terminal kinase) MAPK (mitogen-activated protein kinase) family. We have also defined a minimal region of the cPLA2 promoter that is critical for this induction. To identify potential transcription factors that bind to this region and regulate expression, a yeast one-hybrid screen was performed with a rat lung cDNA library. Multiple members of the Kruppel family were identified, with LKLF (lung Kruppel-like factor) being isolated a number of times. Overexpression of LKLF in lung epithelial cells or Drosophila SL-2 cells increased cPLA2 promoter activity. Conversely, expression of a dominant negative form of LKLF inhibited induction of cPLA2 promoter activity by oncogenic Ras in normal lung epithelial cells and NSCLC. By electrophoretic mobility-shift assay analysis, it was found that LKLF bound to a GC-rich region of the cPLA2 promoter located between -37 and -30 upstream from the transcription start site. Expression of siRNA (small interfering RNA) directed against LKLF inhibited basal expression of cPLA2 in lung epithelial cells and blocked induction by H-Ras. In NSCLC, siRNA against LKLF co-operated with siRNA against Sp1 (stimulatory protein 1) to inhibit cPLA2 promoter activity. Finally, recombinant LKLF was a substrate for ERKs. These results indicate that LKLF is an important regulator of cPLA2 expression and participates in the induction of this protein, which is critical for increased eicosanoid production associated with lung tumorigenesis. PMID- 15540990 TI - Origin and chemical coding of primary afferent neurones supplying the prostate of the dog. AB - Retrograde tracing technique combined with the double-fluorescent immunohistochemistry were used to investigate the distribution and chemical coding of primary afferent neurones supplying the canine prostate. After the injection of Fast Blue (FB) into the prostatic tissue retrogradely-labelled (FB(+)) primary afferent neurones were localized in bilateral L(1)-Ca(1) dorsal root ganglia (DRG). Statistical analysis using anova test showed that there are two major sources of afferent prostate innervation. The vast majority of prostate supplying primary afferent neurones were located in bilateral L(2)-L(4) DRG (56.9 +/- 0.6%). The second source of the afferent innervation of canine prostate were bilateral S(1)-Ca(1) DRG (40.6 +/- 1.0%). No statistically significant differences were found between average number of FB(+) neurones localized in the left and right DRG (49.5 +/- 1.7 and 50.5 +/- 1.7%, respectively). Immunohistochemistry revealed that FB(+) primary afferent neurones contain several neuropeptides in various combinations. In the prostate-supplying neurones of lumbar and sacro-caudal DRG the immunoreactivity to substance P (SP) and calcitonin gene-related peptide (CGRP) was found most frequently (50 +/- 3.7 and 37.3 +/- 1.9%, respectively). Both in the lumbar and sacro-caudal DRG, considerable population of FB(+) neurones immunoreactive neither to SP nor CGRP were also found (23 +/- 2.6 and 32.8 +/- 2.3%, respectively). In the lumbar DRG 10.7 +/- 1.1% of SP-immunoreactive FB(+) neurones also contained galanin (GAL). In 9.2 +/- 2.2% of the prostate-supplying primary afferent neurones located in the sacro-caudal DRG the co-localization of SP and GAL was also reported. Results of the retrograde tracing experiment demonstrated for the first time sources of afferent innervation of the canine prostate. Double immunohistochemistry revealed that many of the prostate-supplying primary afferent neurones express some of sensory neuropeptides which presumably may be involved in nociception and some pathological processes like inflammation or nerve injury. PMID- 15540991 TI - The effect of vascular endothelial growth factor on in vitro embryonic heart development in rats. AB - In vitro effects of vascular endothelial growth factor (VEGF) on heart development and total embryonic growth were investigated in 84 rat embryos (obtained from nine pregnant females) at 9.5 days of gestation that were cultured in whole rat serum (WRS), in <30 kDa + >50 kDa serum fractions [retenate (R)], and in R + VEGF. After 24-h culture, the embryos from each group were harvested and divided into two groups. One group was analysed morphologically and biochemically to obtain embryo protein content, the second group was serially sectioned and examined by light microscopy. Morphological score, embryo protein content, somite number and crown-rump length of embryos indicated that embryos cultured in R had significant embryonic retardation, whereas the addition of VEGF to R increased embryonic growth and development. The morphological scores for WRS, R and R + VEGF were 57.7 +/- 0.87, 46.6 +/- 1.90 and 52.1 +/- 0.97, somite numbers were 26.5 +/- 0.47, 20.1 +/- 0.63 and 24.4 +/- 0.46, crown-rump lengths were 3 +/- 0.07, 2.4 +/- 0.06 and 2.7 +/- 0.06 mm, and embryo protein contents were 160.5 +/- 7.41, 98.2 +/- 4.81 and 141.1 +/- 10.96 mug per embryo, respectively. The results of histological examination of heart development were similar. The hearts of embryos grown in R were unseptated and tubular. The atrioventricular endocardial cushions were incompletely developed. The addition of VEGF to R improved heart development. There were no gross morphological differences in the cardiac development between embryos grown in WRS and R + VEGF. In both groups, development of the muscular interventricular septum had begun. Development of the atrioventricular cushions was also similar in both groups and had caused narrowing of the atrioventricular canals, but the atrial septation was not observed. PMID- 15540992 TI - The subchondral split line patterns of the medial coronoid process in canine ulnae. AB - The subchondral split line patterns of the canine medial coronoid process (MCP) were compared with fragmentation line patterns of the MCP in case of elbow dysplasia. Split line patterns were determined in paired ulnae from the killed 26 large-breed dogs ranging in age between 0.8 and 15 years and in two ulnae affected by fragmentation of the MCP. The macerated ulnae were degreased with methylene chloride and decalcified in 5% nitric acid. The subchondral bony layer was pierced at right angles in regular intervals using a round needle that was dipped in black liquid acrylic colour. Three main types of split line patterns could be differentiated (i.e. a sagittal type, a transverse type and an intermediate type). In the sagittal type, split lines were aligned in parallel to the lateral border and at right angles to the rim of the tip and medial border of the MCP. In the transverse type, split lines were orientated in a transverse line to both collateral borders. The intermediate type was characterized as a transition type between sagittal and transverse type as the split lines were aligned obliquely to the longitudinal axis of the MCP. These three types corresponded well with the fissure and fragmentation line patterns of the MCP. The present findings strongly suggest an association between split line pattern and type of fragmentation of the MCP. PMID- 15540993 TI - Immunohistochemical localization of S-100 protein in the pig pituitary gland. AB - Immunohistochemical localization of S-100 protein was studied in anterior, intermediate and posterior lobe of the pig pituitary gland. Two immunopositive cells for S-100 protein were identified: the folliculo-stellate cells (FSc) in the glandular lobes and the pituicytes in the neural lobe. In the anterior lobe, immunoreactive folliculo-stellate cells were scattered among secretory cells. In the area where the secretory cells form strands and follicle-like groups the positive cells were concentrated in groups. In the intermediate lobe, S-100 protein-positive cells were located sparsely among secretory cells and next to secretory follicles and the pituitary cleft. These FSc were more voluminous and displayed fewer cytoplasmic processes. In the neurohypophysis, positive reaction for S-100 protein was seen in the pituicytes. These cells were distributed singly or concentrated in groups. The distribution, and morphologic characteristics of the FSc in the glandular lobes and the pituicytes in the neural lobe in the pig indicate different origin of both S-100 protein-positive cells. PMID- 15540994 TI - Decrease of nuclear reactivity to growth-regulatory galectin-1 in senescent human keratinocytes and detection of non-uniform staining profile alterations upon prolonged culture for galectin-1 and -3. AB - Summary Multipotent stem cells (source for interfollicular epidermis, hairs and sebaceous glands) are localized in the bulge region of the outer root sheath of hair follicles, while stem cells giving rise to interfollicular epidermis reside in its basal. Using the multifunctional lectin galectin-1 as a marker to localize accessible binding sites in situ as a step to figure out galectin functionality in stem cells, we studied hair follicle-derived keratinocytes. Specific nuclear binding of galectin-1 associated with expression of DeltaNp63alpha, a potential marker of epidermal stem cells, was detected. Binding of chimera-type galectin-3 to a nuclear site was not found in parallel assays. During the process of ageing in culture when cells acquire properties of senescence, disappearance of the nuclear signal for galectin-1 binding was accompanied by a similar decrease of nuclear DeltaNp63alpha expression and increased binding of galectin-3 to the cell membrane, namely in regions of intercellular contacts. Expression of cytokeratin 10, a marker of the terminal differentiation was seen only in a small fraction of the cell population. These data extend the evidence for nuclear sites with galectin-1 reactivity in squamous epithelial cells, the expression of which is modulated upon senescence. Moreover, the results document the divergence of galectin-1 and -3 on the level of ligand selection in this cell type, underscoring the importance of the technical aspect to employ tissue lectins as probe and to perform a fingerprinting with several markers of the galectin family in parallel. PMID- 15540995 TI - Is the rhesus monkey (Macaca mulatta) comparable to humans? Histomorphology of the sphincteric musculature of the lower urinary tract including 3D reconstruction. AB - The physiology of the muscle systems of the human lower urinary tract is still not known in detail. To study the functional basics of this complex organ system, experiments are often performed in animal models including rhesus monkeys. To apply the results of animal model studies to the humans, a clear knowledge of the comparative anatomy of both species is necessary. However, detailed comparative studies of the lower urinary tract of the rhesus monkey and the humans are lacking. Accordingly, a detailed study on the sphincteric musculature of the lower urinary tract of the rhesus monkey was performed in order to demonstrate anatomical correspondences and differences between both species. The lower urinary tract anatomy was investigated in 18 male and female rhesus monkeys (Macaca mulatta) by serial sections. Immunohistochemical staining methods were used to differentiate striated and smooth musculature. Three-dimensional reconstructions were performed in order to demonstrate the topographical anatomy of the different muscle systems. In both man and male rhesus monkeys, a urethral sphincter muscle exists independent of the pelvic floor musculature, with a smooth and a striated muscular part. A urinary diaphragm (diaphragma urogenitale) does neither exist in the rhesus monkey nor in the human. In contrast to women, a striated muscle encircles the urethra and vagina together in the female rhesus monkey. A vesical sphincter muscle, found in the human bladder outlet, does not exist in the rhesus monkey. PMID- 15540997 TI - There is no evidence of an association in children and teenagers between the apolipoprotein E epsilon4 allele and post-traumatic brain swelling. AB - Traumatic brain injury (TBI) is an important cause of mortality and disability in children and teenagers. A particular feature of the neuropathology at post-mortem is brain swelling. The cause of the swelling in some cases is not known, while in others it is associated with traumatic axonal injury or hypoxia. Apolipoprotein E (APOE) epsilon4 allele is known to be an important genetic determinant of outcome in children after TBI. We hypothesized a relationship between possession of APOEepsilon4 and diffuse traumatic brain swelling. A total of 165 cases aged between 2 and 19 years were identified from the department's tissue archive. APOE genotype was determined by polymerase chain reaction (PCR) in 106 cases. Bilateral swelling was present in 44 cases (11 with APOEepsilon4), unilateral swelling in 25 cases (7 with APOEepsilon4) and in 36 cases (9 with APOEepsilon4) there was no evidence of brain swelling. There was no significant relationship between possession of APOEepsilon4 and the presence of cerebral swelling (chi(2) = 0.09, df = 2, P = 0.96). The 95% confidence interval for difference in proportions with swelling in those with and without the APOE epsilon4 is -19% to 22%. Thus, a significant relationship was not found between diffuse brain swelling and possession of APOEepsilon4, and in this cohort of patients there was an identifying cause of the brain swelling in all cases. PMID- 15540998 TI - The extent of axonal loss in the long tracts in hereditary spastic paraplegia. AB - Hereditary spastic paraplegia (HSP) comprises a group of inherited neurodegenerative disorders with the shared characteristics of progressive weakness and spasticity predominantly affecting the lower limbs. Limited pathological accounts have described a 'dying back' axonal degeneration in this disease. However, the distribution and extent of axonal loss has not been elucidated in a quantitative way. We have studied post-mortem material from six HSP patients and 32 controls in detail. The population of axons was examined quantitatively in the corticospinal tracts from the medulla to the lumbar spinal cord and the sensory tracts from the lumbar to upper cervical spinal cord. Myelin and axon-stained sections were employed to estimate the notional area and axonal density, respectively, of both tracts. Our results indicate that in the corticospinal tracts there is a significant reduction in area and axonal density at all levels investigated in HSP compared to controls. In the corticospinal tracts, the ratio of medulla and lumbar total axonal number was significantly greater in HSP cases compared to controls suggesting more pronounced axonal loss in the distal neuraxis in HSP than in controls. The sensory tracts in HSP, in contrast, showed a significant reduction in area and axonal density only in the upper regions of the spinal cord. Similar to the corticospinal tracts, the ratio of lumbar and upper cervical cord total axonal number in the sensory tracts was increased in HSP cases compared to controls. These findings are consistent with a length-dependent 'dying back' axonopathy. Nerve fibre loss was not size-selective with both small and large diameter fibres affected. In HSP, axonal loss is widespread and symmetrical and its extent tract-specific. The characterization of the nature of axonal loss in HSP, where this is a primary phenomenon, may help the interpretation of axonal loss in conditions such as multiple sclerosis where the sequence of events is less clear. PMID- 15540999 TI - Transforming growth factor-betas in a rat model of neonatal posthaemorrhagic hydrocephalus. AB - Posthaemorrhagic ventricular dilatation (PHVD) is a common complication of intraventricular haemorrhage in premature infants. The aim of this study was to investigate the role of transforming growth factor-betas (TGF-betas), a family of polypeptides with potent desmoplastic properties, in the aetiology of PHVD in a newly developed neonatal rat model of this disorder. Pups were injected with citrated rat blood or artificial cerebrospinal fluid (ACSF) into alternate lateral ventricles on postnatal days 7 and 8. The brains were perfusion-fixed 14 days later and immunohistochemistry was performed for TGF-beta1, -beta2 and beta3, p44/42 mitogen-activated protein (MAP) kinases, and the extracellular matrix proteins laminin, vitronectin and fibronectin. Ventricular dilatation occurred in 58.3% of animals injected with blood and 36.7% of those injected with ACSF. Periventricular immunoreactivity for TGF-beta1 and -beta2 increased in injected animals irrespective of the presence or absence of ventricular dilatation, although the levels of both isoforms tended to be higher in animals with hydrocephalus. TGF-beta3 immunoreactivity was elevated in hydrocephalic rats only. The immunolabelling for phosphorylated p44/42 MAP kinases rose in a pattern similar to that for TGF-beta1 and -beta2. Expression of TGF-betas was accompanied by deposition of the extracellular matrix proteins fibronectin, laminin and vitronectin. The changes caused by injection of ACSF were the same as those caused by injection of blood. Our results raise the possibility that expression of TGF-betas, together with extracellular matrix protein deposition, may be involved in the development and/or maintenance of hydrocephalus after ventricular distension due to haemorrhage in the neonate. PMID- 15541000 TI - Differential expression of alpha-synuclein isoforms in dementia with Lewy bodies. AB - Dementia with Lewy bodies (DLB) is characterized by the widespread presence of Lewy bodies (LBs) in the brain. alpha-Synuclein, the main component of LBs, is expressed as two main isoforms (112 and 140), but little is known about their differential expression in the brain. We compared alpha-synuclein 112 and alpha synuclein 140 expression levels in the prefrontal cortices of six DLB patients, eight Alzheimer disease (AD) patients, and six control subjects. Relative alpha synuclein 112 and alpha-synuclein 140 expression levels were determined by real time polymerase chain reaction with competimer technology using a LightCycler System. Whereas total alpha-synuclein levels were just marginally elevated in DLB in comparison with the other groups, alpha-synuclein 112 was seen to be markedly increased in DLB compared with AD cases and controls. In contrast, alpha synuclein 140 levels were significantly diminished in both neurodegenerative disorders in comparison with controls. These results show differential overexpression of alpha-synuclein 112 in DLB, a finding that could be of importance in DLB pathogenesis. PMID- 15541001 TI - Presence of D110 antigen expressing immunocompetent cells in glioblastoma associates with prolonged survival. AB - Monoclonal antibody D110 has recently been described as a novel marker of hypoxic tissue damage, reacting with a so far unknown antigen with preferential expression in the central nervous system. The aim of the study was to investigate D110 immunoreactivity in glioblastoma, its association with the expression of hypoxia-related proteins and its impact on patient outcome. A total of 114 consecutive adult patients who underwent first operation of primary glioblastoma were included in this study. We evaluated D110 immunoreactivity qualitatively and semi-quantitatively and correlated it with expression of hypoxia inducible factor 1 alpha (HIF-1alpha), expression of vascular endothelial growth factor (VEGF), and with patient survival using univariate and multivariate statistical analysis. We observed D110 immunolabelling in 85.1% of the cases. D110 immunoreactivity was detectable in infiltrating HLA-DR and CD68 expressing cells, most likely microglial cells or haematogenous cells of monocyte/macrophage lineage. In the peripheral lymphoreticular system, immunohistochemistry disclosed selective D110 labelling of Langerhans cells and of dendritic cells of the thymic medulla. Univariate statistical analysis revealed significantly longer survival of patients whose glioblastomas contained D110 immunoreactive infiltrating cells. There was no association between presence of D110 immunoreactive cells and expression of HIF-1alpha and VEGF. We conclude that D110 immunoreactivity in glioblastoma does not seem to be related to tissue hypoxia. D110 identifies immunocompetent cells, which positively influence survival of glioblastoma patients. PMID- 15541002 TI - Depletion of MAP2 expression and laminar cytoarchitectonic changes in dorsolateral prefrontal cortex in adult autistic individuals. AB - The neuropathological substrates underlying the characteristic clinical phenotype of autism are unknown. Neuroimaging studies have identified a decrease in task related activation in the dorsolateral prefrontal cortex in autism. In the current study, we have analysed the dorsolateral prefrontal cortex in two adult individuals with a clinical diagnosis of autism, using Nissl staining and MAP2 immunohistochemistry. There was unchanged density of both neuronal and glial cell pools, although the autistic individuals had ill-defined neocortical cellular layers, substantially depleted MAP2 neuronal expression, and reduced dendrite numbers. Further studies on a larger number of individuals with autism are needed to establish the clinical relevance of the described changes, especially to determine whether the loss of dendritic markers is age associated or disease specific. PMID- 15541003 TI - Tau aggregates are abnormally phosphorylated in inclusion body myositis and have an immunoelectrophoretic profile distinct from other tauopathies. AB - Sporadic inclusion body myositis (s-IBM) is the most frequent progressive acquired inflammatory myopathy in people older than 50 years. Abnormal aggregates of 'Alzheimer's proteins', including tau proteins, have been previously demonstrated in s-IBM. In the present study, we have investigated by immunohistochemistry and immunoblotting analysis the presence of tau proteins in muscle biopsy samples from patients with s-IBM and other myopathies with rimmed vacuoles, using newly developed antibodies raised against tau protein epitopes found in Alzheimer's disease brain. Tau immunoreactivity was shown in rimmed vacuoles or inclusions, preferentially with antibodies directed against phosphorylated carboxy-terminal epitopes of tau proteins. Cytoplasmic reactivity was also demonstrated in atrophic, nonvacuolated fibres, as well as in non necrotic fibres invaded by inflammatory cells. Abnormally phosphorylated tau aggregates were also found in other compartments of the muscle fibre in s-IBM and other myopathies. Tau immunoblotting showed an electrophorectic profile of a doublet within the range of 60-62 kDa isovariants, which was different from tauopathies of the central nervous system. Finally, the unique pattern of immunoreactivity of s-IBM samples towards anti-tau antibodies is a new clue to a possible distinct subclass of peripheral tauopathy, different from the tauopathies of the central nervous system. PMID- 15541004 TI - Alterations in neurofilament protein(s) in human leprous nerves: morphology, immunohistochemistry and Western immunoblot correlative study. AB - Using a specific antibody (SMI 31), the state of phosphorylation of high and medium molecular weight neurofilaments (NF-H and NF-M) was studied in 22 leprous and four nonleprous human peripheral nerves by means of immunohistochemistry, sodium dodecyl sulfate-poly acrylamide gel electrophoresis (SDS-PAGE) and Western immunoblot (WB). The results thus obtained were compared with morphological changes in the respective nerves studied through light and electron microscopy. Many of the leprous nerves showing minimal pathology revealed lack of or weak staining with SMI 31, denoting dephosphorylation. Remyelinated fibres stained intensely with SMI 31 antibody. The WB analysis of Triton X-100 insoluble cytoskeletal preparation showed absence of regular SMI 31 reactive bands corresponding to 200 and 150 kDa molecular weight (NF-H and NF-M, respectively) in 10 nerves. Three of the 10 nerves revealed presence of NF protein bands in SDS PAGE but not in WB. Presence of additional protein band (following NF-M) was seen in four nerves. Two nerves revealed NF-H band but not NF-M band and one nerve showed trace positivity. In the remaining five nerves presence of all the three NF bands was seen. Thus, 77.3% (17/22) of human leprous nerves studied showed abnormal phosphorylation of NF protein(s). The ultrastructural study showed abnormal compaction and arraying of NF at the periphery of the axons in the fibres with altered axon to myelin thickness ratio (atrophied fibres) as well as at the Schmidt-Lantermann (S-L) cleft region. Such NF changes were more pronounced in the severely atrophied axons suggesting a direct correlation. The observed well-spaced NF in the remyelinated fibres under ultrastructural study was in keeping with both intense SMI 31 staining and presence of NF triplet bands seen in WBs in four of leprous nerves that showed a large number of regenerating fibres suggesting reversal of changes with regeneration. Findings in the present study suggest that atrophy, that is, the reduction in axonal calibre and paranodal demyelination, seen in leprous nerves may result from dephosphorylation of NF-H and NF-M proteins. PMID- 15541005 TI - Neurotrophin receptor immunoreactivity in the hippocampus of patients with mesial temporal lobe epilepsy. AB - Recent evidence supports a critical role of neurotrophins in the regulation of both neuronal survival and synaptic transmission during epileptogenesis. We have examined the immunohistochemical expression of high- (tyrosine kinase receptors, trk) and low-affinity (p75) neurotrophin receptors (NTRs) in the hippocampal specimens from 18 patients with chronic temporal lobe epilepsy [TLE; 14 patients with hippocampal sclerosis (HS) and four with focal lesions (tumours) not involving the hippocampus proper]. Nonepileptic autopsy brains (n = 6) and surgical specimens from tumour patients without epilepsy (n = 3) were used as controls. Immunoreactivity (IR) for the trk receptors (trkA, trkB, trkC) was detected in normal human brain within the pyramidal neurones of hippocampal cornus ammoni (CA) regions and in the dentate gyrus. There were no detectable differences in the neuronal trk IR patterns in the hippocampus between control and TLE cases with HS, except for a decrease in neuronal density in regions where cell death had occurred (CA1, CA3 and CA4). In contrast, a consistent increase in trkA IR was observed in reactive astrocytes in CA1 and dentate gyrus. The low affinity p75 neurotrophin receptor (p75(NTR)) was expressed in low levels in postnatal normal hippocampus. In contrast, neuronal p75(NTR) IR was detected in 10/14 cases of HS in spared neurones within the CA and hilar regions of the hippocampus. Double labelling revealed that p75(NTR)-positive neurones also contain trk receptor IR. In six cases with prominent glial activation strong p75(NTR) IR was observed in microglial cells within the sclerotic hippocampus. The present results indicate that changes in NTR expression are still detectable in the hippocampus of patients with chronic TLE and involve both glial and neuronal cells. Reactive astrocytes were immunoreactive for trkA, whereas activated microglia cells were reactive for p75(NTR), suggesting different functions for specific NTRs in the development of reactive gliosis. Moreover, the increased expression of p75(NTR) in hippocampal neurones of TLE patients may critically influence the neuronal survival during the epileptogenic process. PMID- 15541006 TI - Sharing of polyglutamine localization by the neuronal nucleus and cytoplasm in CAG-repeat diseases. AB - The expansion of a trinucleotide cytosine adenine and guanine (CAG) repeat that codes for polyglutamine is a common gene mutation in the family of hereditary neurodegenerative diseases that includes Machado-Joseph disease (MJD) and dentatorubral-pallidoluysian atrophy (DRPLA). The presence of ubiquitinated neuronal intranuclear inclusions (NIIs) has been recognized as a neuropathological hallmark of these diseases, although the significance of NIIs in the pathogenesis remains a matter of controversy. In a previous study of DRPLA, we proposed that intranuclear diffuse accumulation of mutant proteins is another pathological characteristic of neurones, and that the variable prevalence of this characteristic may be relevant to the variation of clinical symptoms in patients with different repeat sizes. Recently, we also disclosed that polyglutamine tracts are localized in a subset of lysosomes in affected neurones. The present immunohistochemical study of autopsied MJD and DRPLA brains shows that the nucleus and cytoplasm of affected neurones share the subcellular distribution of expanded polyglutamine tracts, the pattern of distribution being specific to each diseased brain. The results suggest that in CAG-repeat diseases, mutant proteins are involved in both the ubiquitin/proteasome and endosomal/lysosomal pathways for protein degradation in different intraneuronal compartments, where their accumulation may exert distinct influences on neuronal physiology. PMID- 15541007 TI - Ballooned neurones in the limbic lobe are associated with Alzheimer type pathology and lack diagnostic specificity. AB - Ballooned neurones (BNs) are one of the pathological hallmarks of several neurodegenerative diseases, including Pick's disease, corticobasal degeneration and argyrophilic grain disease (AGD). They have also been described in Alzheimer disease (AD), but the frequency of BNs in AD has not been systematically addressed. In the present study, immunohistochemistry for alphaB-crystallin was used as a sensitive method to detect BNs to determine the frequency of BNs in the limbic lobe in AD. At least a few BNs were detected in the limbic lobe of virtually all AD cases, and their density correlated with Braak stage, as well as the density of neurofibrillary tangles and senile plaques in the limbic lobe. The density of BN tended to be greater in AD cases with concurrent AGD than in pure AD. Given the high prevalence of AD in brain banks for neurodegenerative disease and the frequent presence of BNs in these areas with alphaB-crystallin immunohistochemistry, the present findings further indicate that BNs confined to the limbic lobe lack specificity in diagnostic neuropathology. PMID- 15541008 TI - Up-regulation of a growth arrest and DNA damage protein (GADD34) in the ischaemic human brain: implications for protein synthesis regulation and DNA repair. AB - GADD34 is a growth arrest and DNA damage inducible gene up-regulated in response to DNA damage, cell cycle arrest and apoptosis. It is thought that GADD34 may play a crucial role in cell survival in ischaemia. GADD34 expression was assessed immunohistochemically in post-mortem human hippocampal tissue obtained from patients surviving for defined periods (0-24 h; 24 h-7 days) after a cardiac arrest and in age-matched control subjects. In control brain, cytoplasm staining in GADD34 immunopositive cells was faint but present throughout the hippocampus and cortex. There was minimal change in GADD34 expression in the group surviving 0-24 h after cardiac arrest. However GADD34 immunostaining was markedly increased in selectively vulnerable regions in the 24 h-7 day survival group. Increased GADD34 staining was present in ischaemic neurones and in some morphologically normal neurones after cardiac arrest. Extensive ischaemic damage was found to correlate with elevated GADD34 immunostaining in the CA1 layer of the hippocampus (**P < 0.0016). In addition, GADD34 was found to colocalize with proliferating cell nuclear antigen in some neurones. The up-regulation of GADD34 in response to global ischaemia in the human brain plus its influence on protein synthesis and DNA repair suggests that this protein may have the potential to influence cell survival. PMID- 15541009 TI - Inflammatory myofibroblastic tumour of the sella turcica. PMID- 15541010 TI - Malignant intracranial solitary fibrous tumour with four recurrences over a 30 year period. PMID- 15541012 TI - Differential effects of FK506 and cyclosporin A on catecholamine release from bovine adrenal chromaffin cells. AB - 1 The effects of the immunosuppressants, tacrolimus (FK506) and cyclosporin A (CsA), on catecholamine (CA) release were examined in cultured bovine adrenal chromaffin cells. 2 In intact cells, FK506 (1-30 microM) inhibited CA release stimulated by acetylcholine (ACh; 100 microM), 1,1-dimethyl-4-phenyl-piperazinium (DMPP, 10 microM) or high K+ (40 mM). CsA (1-30 microM) had a little inhibitory effect on the ACh- or DMPP-stimulated CA release, whereas it enhanced the high K(+)-stimulated CA release. 3 In beta-escin-permeabilized cells, FK506 inhibited CA release stimulated by Ca2+ (1 and 10 microM) in the presence and absence of MgATP (2 mM). CsA induced CA release under Ca(2+)-free condition and enhanced the Ca(2+)-stimulated CA release in the presence and absence of MgATP. 4 It is known that the Ca(2+)-dependent exocytosis involves at least two distinct steps, ATP requiring priming stage and ATP-independent fusion step in adrenal chromaffin cells. Therefore, it is suggested that FK506 inhibits the Ca(2+)-dependent exocytosis probably at the fusion step whereas CsA induces CA release from bovine adrenal chromaffin cells. PMID- 15541013 TI - Actions of R- and S-verapamil and nifedipine on rat vascular and intestinal smooth muscle. AB - 1 We have investigated the actions of the calcium entry blockers nifedipine, R verapamil and S-verapamil in rat aorta, colon and vas deferens. 2 In aorta and colon, these agents produced concentration-dependent relaxations of KCl (80 mM) induced contractions. In both tissues, the order of potency was nifedipine > S verapamil > R-verapamil. However, nifedipine showed selectivity for aorta (potency ratio, colon/aorta: 4.36), S-verapamil showed no selectivity (0.62), but R-verapamil showed selectivity for colon (0.19). 3 In prostatic portions of rat vas deferens, nifedipine (10 microM) abolished the contraction to a single electrical stimulus, but R- and S-verapamil were without effect. In epididymal portions of rat vas deferens, R- and S-verapamil inhibited alpha1-adrenoceptor mediated contractions to a single electrical stimulus at concentrations of 10 microM and above. 4 In conclusion, R-verapamil may prove useful as an intestinal selective calcium entry blocker in the treatment of intestinal disease with a hypermotility component, e.g. irritable bowel syndrome. PMID- 15541014 TI - Effects of atipamezole--a selective alpha-adrenoceptor antagonist--on cardiac parasympathetic regulation in human subjects. AB - 1 This double-blind, cross-over, placebo-controlled study on six healthy male volunteers was designed to evaluate the effects of alpha2-adrenoceptor antagonism on cardiac parasympathetic regulation. 2 The subjects received atipamezole intravenously as a three-step infusion, which aimed at steady-state serum concentrations of 10, 30 and 90 ng ml(-1) at 50-min intervals. 3 Drug effects were assessed with repeated recordings of blood pressure and electrocardiogram, in which the high-frequency (0.15-0.40 Hz) R-R interval variation is supposed to reflect cardiac parasympathetic efferent neuronal activity. 4 At the end of the three steps of the infusion, the mean (+/-SD) concentrations of atipamezole were 10.5 (3.9), 26.8 (5.6) and 81.3 (21.1) ng ml(-1). 5 Within this concentration range, atipamezole appeared to reduce slightly the high-frequency R-R interval fluctuations, indicating a minor vagolytic effect in the heart. 6 Atipamezole increased systolic and diastolic arterial pressure, on average by 20 and 14 mmHg (maxima at the second step of the infusion), which evidently reflects an overall sympathetic augmentation. PMID- 15541015 TI - Comparison of noradrenaline and lysosphingolipid-induced vasoconstriction in mouse and rat small mesenteric arteries. AB - 1 We have compared vasoconstriction responses in isolated mesenteric small arteries from mice and rats as elicited by KCl, noradrenaline and the lysosphingolipids sphingosine-1-phosphate (S1P) and sphingosylphosphorylcholine (SPC). 2 Contractile responses to KCl and noradrenaline, but not those of S1P or SPC, were significantly related to vessel diameter in both species. 3 When comparing vessels of similar diameter, contractile responses for KCl and the three agonists were much smaller in mice than in rats, e.g. 8.3 +/- 0.4 vs. 14.7 +/- 0.7 mn for noradrenaline. 4 Based upon the antagonist rank order of potency of prazosin (pKB 8.80) > B8805-033 (pKB 7.89) > yohimbine (pKB 6.18) approximately BMY 7378 (pA2 6.03), noradrenaline responses in mice were mediated solely via alpha1A-adrenoceptors, similar to what repeatedly has been shown in rats. 5 The S1P3 receptor antagonist suramin (100 microM) significantly inhibited responses to S1P and SPC in rats but not in mice, and did not affect noradrenaline responses in either species. 6 We conclude that for any given diameter, mouse mesenteric arteries develop less contraction in response to various stimuli. Noradrenaline acts via alpha1A-adrenoceptors in both species. Responses to S1P and SPC differ between both species with regard to suramin sensitivity indicating involvement of different receptor subtypes for lysosphingolipids in both species. PMID- 15541017 TI - In memoriam-Kiyoshi Toda, MD (1931-2004). PMID- 15541018 TI - The neuromelanin of human substantia nigra: physiological and pathogenic aspects. AB - Neuromelanin (NM) accumulates as a function of age in normal human substantia nigra (SN) but is relatively depleted in the SN of patients with Parkinson disease (PD). Several studies have been performed to further our understanding of the role of NM in neuronal aging and neurodegenerative mechanisms of PD. To this purpose, NM from human SN was isolated and its structure and molecular interactions were investigated. Cysteinyl-dopamine was shown to be one precursor of NM synthesis. A striking affinity of NM for specific metals, lipids, drugs and pesticides was found in vitro, and in animal and human brain postmortem studies. Because of these affinities, NM seems to play a protective role in the human brain by blocking toxic molecules. On the other hand, experiments in cell culture indicate that NM can activate microglia, eliciting the release of cytotoxic factors that can induce neurodegeneration. PMID- 15541019 TI - Non-invasive measurements of skin pigmentation in situ. AB - Objective in situ measurements of skin pigmentation are needed for accurate documentation of pigmentation disorders, in studies of constitutive and induced skin pigmentation, for testing of the efficacy of pro-pigmentation or de pigmentation agents, etc. Non-invasive instrumental measurements of skin pigmentation have been used for many decades. All are based on the ability of melanin to attenuate light. However, hemoglobin in dermal capillaries also attenuates light and needs to be accounted for when pigmentation is assessed. The methods under consideration include: (a) single point measurements, in which light reflected from a defined skin area is collected and a pigment index is calculated representing the average pigmentation over the examined area, and (b) imaging methods that attempt to generate a concentration distribution map of melanin pigment for the skin area being imaged. In this article, we describe the potentials and the limitations of the different approaches to both single point and imaging methods. PMID- 15541020 TI - The melanocortin receptor-1 gene but not the proopiomelanocortin gene is expressed in melanoblasts and contributes their differentiation in the mouse skin. AB - Alpha-melanocyte stimulating hormone (alpha-MSH) added to serum-free primary culture of melanoblasts derived from epidermal cell suspensions of 0.5 d old C57BL/10JHir mice induced their differentiation. Analysis using the reverse transcription-polymerase chain reaction showed that the expression of the melanocyte-specific alpha-MSH receptor gene, melanocortin receptor-1 (MC1-R), had already been initiated before addition of alpha-MSH, and, in addition, no up regulation of the MC1-R gene was observed after addition of alpha-MSH. However, no expression of the proopiomelanocortin (POMC) gene was observed before or after the addition of alpha-MSH. The expression of the MC1-R and POMC genes in the epidermis and dermis of the dorsal skin was surveyed from 13 d old embryos to 5.5 d old neonates. The expression of the MC1-R gene was first observed in the epidermis of 13 d old embryos, and gradually increased up to 0.5 d after birth, and thereafter remained constant. By contrast, the expression of the MC1-R gene in the dermis was first observed in 16 d old embryos, and gradually increased up to 3.5 d after birth, and thereafter remained constant. However, no expression of the POMC gene was observed in the epidermis or dermis of the dorsal skin at any age of mice tested. These results suggest that the expression of the MC1-R gene, but not of the POMC gene, plays an important role in the regulation of melanocyte differentiation in mouse skin. PMID- 15541021 TI - Uveal melanocytes produce matrix metalloproteinases-2 and -9 in vitro. AB - The purpose of the present study was to investigate the expression of matrix metalloproteinase (MMP)-2 and MMP-9 by cultured human uveal melanocytes, and to test the effects of 12-O-tetradecanoyl-phorbol-13-acetate on the expression of these MMPs. Gelatin zymography of conditioned culture medium from four cultures of human uveal melanocytes (two cultures of iridal melanocytes and two cultures of choroidal melanocytes) detected MMP-2 (72 kDa) and a relatively small amount of MMP-9 (92 kDa), both in the latent form. RT-PCR analysis revealed the MMP-2 mRNA and MMP-9 mRNA in cultured uveal melanocytes. Addition of 12-O-tetradecanoyl phorbol-13-acetate (10 ng/ml) to the culture medium caused an increase of production of MMP-2 and MMP-9 by cultured uveal melanocytes, and also stimulated the transcription of MMP-2 and MMP-9 of these cells. PMID- 15541022 TI - Establishment by an original single-cell cloning method and characterization of an immortal mouse melanoblast cell line (NCCmelb4). AB - We devised a unique new single-cell cloning method which uses microscope cover glasses and established a melanoblast cell line derived from mouse neural crest cells. A microscope cover glass was nicked and broken into small pieces and put on a dish. Culture medium and a suspension of 20-30 cells/ml were dropped in the dish. After 1-3 d, a piece of glass to which only one cell was adhered was picked up and transferred to another dish containing culture medium. The greatest advantage of this method is that the derivation of a colony from a single cell can be directly confirmed by microscopy and there is no risk of migratory cells being contaminated by other colonies. Using this single-cell cloning method, in this study we established a cell line derived from a neural crest cell line (NCC S4.1) and designated it as NCCmelb4. When the culture medium was supplemented with stem cell factor (SCF) alone, NCCmelb4 cells were KIT-positive and tyrosinase-negative melanocyte precursors; they remained at an immature and undifferentiated stage. When the medium was supplemented with phorbol 12-o tetradecanoyl-13-acetate (TPA) + cholera toxin (CT), the cell morphology changed and became L-3,4-dihydroxyphenylalanine (DOPA)-positive. This observation indicates that the NCCmelb4 cells are capable of further differentiation with suitable stimulation. NCCmelb4 cells derived from the mouse neural crest has characteristics of melanocyte precursors (melanoblasts), and is a cell line which can be utilized to study differentiation-inducing factors and growth factors without the effects of feeder cells. PMID- 15541023 TI - Pigmentation in intrinsically aged skin of A1 guinea pigs. AB - It is known that skin often shows irregular pigmentation during aging which is frequently associated with hyperpigmentation. Many studies have utilized brownish A1 guinea pigs to investigate the pathogenesis of ultraviolet (UV)-induced skin pigmentation, however, changes associated with intrinsic aging in A1 guinea pig skin have not been documented. To characterize such changes, skin from the dorsal and neck areas of 20-week, 1-, 2-, 3- and 5-yr-old guinea pigs was examined. Skin color was measured using a colorimeter, and biopsy specimens were stained with Masson-Fontana, L-3,4-dihydroxyphenylalanine (DOPA), and antibodies against KIT (ACK-45), gp100 (HMB-45) and S-100 proteins. The L* value of skin color decreased with aging and melanin deposits increased in the epidermis. Further, DOPA+, gp100+ and S-100+ melanocytes increased, indicating that the number of melanocytes had increased with age, whereas KIT+ melanocytes did not increase in dorsal skin and actually decreased in neck skin with aging. Further, rippled pigmented areas appeared in the neck skin of the 3-yr-old animals, and in the dorsal and neck skin of 5-yr-old guinea pigs in the absence of UV irradiation. Melanocytes were distributed uniformly in younger skin, whereas they were clustered in older skin. UV irradiation caused an increase in the number of melanocytes, although they were not clustered. These results are the first to provide evidence that pigmentation is induced in the skin of intrinsically aged A1 guinea pigs in the absence of UV irradiation, a process that differs from that elicited by UV irradiation. PMID- 15541024 TI - All trans-retinoic acid (ATRA) elevated eukaryotic translation initiation factor 4A1 (eIF4A1) mRNA in ATRA-responsive vitiliginous epidermis. AB - Chronic skin disorders that require long-term treatment with corticosteroids, such as vitiligo, may use a combination of topical corticosteroids and topical all-trans-retinoic acid (ATRA) to prevent corticosteroid-induced skin atrophy. Besides protecting against the side effects of corticosteroids, ATRA produces a better clinical outcome in some patients. This study examined whether ATRA influences the expression of mRNAs responsible for the clinical correlation. Differential display was performed using kits incorporating an annealing control primer. Epidermis from suction blisters taken from six patients diagnosed with a generalized type of vitiligo, who were included in a placebo-controlled paired comparison left-right study using ATRA and vehicle for 3-6 months, were used. Ten differentially expressed mRNAs were identified in those six patients. Expression levels were restored to normal particularly in four types of mRNAs, which were matched with sequences encoding eukaryotic translation initiation factor 4A1 (eIF4A1), ribosomal protein L13, mediator of RNA polymerase to transcription (MRT) and ribosomal phosphoprotein PO. Of those mRNAs, the level of eIF4A1 mRNA showed a clinical correlation; The expression of eIF4A1 mRNA, examined by real time PCR, was elevated in four patients who showed a favorable clinical response to ATRA, whereas no change or a decrease occurred in three patients whose clinical responses did not differ between ATRA and vehicle treatment. The eIF4A1 protein expression from the other two patients, one of them with a favorable response to ATRA, also showed a clinical correlation. Therefore, eIF4A1 mRNA may be an important gene related to ATRA effects, although further studies are required. PMID- 15541025 TI - Isolation and culture of amelanotic melanocytes from human hair follicles. AB - We report a method to establish amelanotic melanocytes (AMMC) in culture and we investigate the effects of various components in the culture medium. Normal human scalp from cadaver donors was transected 1 mm below the epidermis, and hair follicles in the remaining dermis were isolated by a two-step enzyme treatment. The individual hair follicles were washed exhaustively and suspensions of hair follicle cells were prepared and cultured in Eagle's minimum essential medium supplemented with 12-o-tetradecanoyl-phorbol-13-acetate (TPA), cholera toxin and keratinocyte serum-free medium (K-SFM). Geneticin was used to eliminate contaminating fibroblasts. Proliferation of AMMC was observed after addition of TPA and K-SFM including bovine pituitary extract (BPE) into the culture medium. Cell type was determined by staining with monoclonal antibodies, NKI/beteb and HMB-45, which recognize premelanosomal and melanosomal antigens, respectively. The AMMC were also examined using transmission electron microscopy. Treatment with geneticin eliminates the majority of fibroblasts and does not impair the growth of keratinocytes or AMMC. After contaminating fibroblasts and keratinocytes were removed, two distinct cell morphologies remained: (1) large, dendritic and deeply pigmented cells, which did not proliferate and which disappeared by the third passage, and (2) small bipolar cells, which initially were unpigmented and proliferated very rapidly. We observed that TPA at various concentrations stimulated the proliferation of the cells, and at high concentrations could induce the formation of multiple dendrites. K-SFM including BPE accelerated the proliferation of the cells in a dose-dependent manner. After passage 3, almost all cells expressed premelanosomal and melanosomal antigens, recognized by NKI/beteb and HMB-45, respectively. Active mitochondria, abundant rough endoplasmic reticulum, Golgi complexes, ribosomes and melannosomes (predominantly in stages I, II or III with some at stage IV in some AMMC) were observed ultrastructurally in the cytoplasm of the cultured cells. PMID- 15541026 TI - Comparative histopathology of grey-horse-melanoma and human malignant melanoma. AB - Equine melanoma shows striking features particularly with regard to clinical development in grey horses: in contrast to malignant melanoma in humans and in solid coloured horses that are characterized by early onset of metastasis, pigment cell tumours display almost benign clinical features in ageing grey horses. Through evolution, grey horses appear to be in a favourable position in regard to the biological behaviour of melanomas. Yet unknown factors inhibiting or retarding early melanoma metastasis may be responsible for this phenomenon. In this study, immunostaining profiles and histopathologic patterns of equine vs. human melanotic tumours were compared. In addition, the expression of melanoma markers currently used in human melanoma detection and characterization were evaluated for their applicability in equine melanoma diagnosis. Immunohistopathologic investigations revealed that benign grey horse melanomas share common features with human blue nevi and with human malignant desmoplastic melanomas, whereas their resemblance to other types of human cutaneous malignant melanomas is less pronounced. Our data equally underline that S-100, proliferating cell nuclear antigen (PCNA), HMB-45, Ki-67, T-311 and CD44 can serve as reliable markers for horse melanomas. Further investigations aiming at identifying factors retarding metastasis in affected grey horses are needed, as they may contribute to the development of novel treatment strategies for human malignant melanoma. PMID- 15541029 TI - Hormonal and immunological mechanisms mediating sex differences in parasite infection. AB - The prevalence and intensity of infections caused by protozoa, nematodes, trematodes, cestodes, and arthropods is higher in males than females. The primary thesis of this review is that immunological differences exist between the sexes that may underlie increased parasitism in males compared to females. Several field and laboratory studies link sex differences in immune function with circulating steroid hormones; thus, the roles of sex steroids, including testosterone, oestradiol, and progesterone, as well as glucocorticoids will be discussed. Not only can host hormones affect responses to infection, but parasites can both produce and alter hormone concentrations in their hosts. The extent to which changes in endocrine-immune interactions following infection are mediated by the host or the parasite will be considered. Although males are more susceptible than females to many parasites, there are parasites for which males are more resistant than females and endocrine-immune interactions may underlie this sex reversal. Finally, although immunological differences exist between the sexes, genetic and behavioural differences may explain some variability in response to infection and will be explored as alternative hypotheses for how differences between the sexes contribute to dimorphic responses to parasites. PMID- 15541030 TI - Natural antibody response to Plasmodium falciparum Exp-1, MSP-3 and GLURP long synthetic peptides and association with protection. AB - A longitudinal study was undertaken in Burkina Faso among 293 children aged 6 months to 9 years in order to determine the correlation between an antibody response to several individual malarial antigens and malarial infection. It was found that the presence of a positive antibody response at the beginning of the rainy season to three long synthetic peptides corresponding to Plasmodium falciparum Exp-1 101-162, MSP-3 154-249 and GLURP 801-920 but not to CSP 274-375 correlated with a statistically significant decrease in malarial infection during the ongoing transmission season. The simultaneous presence of an antibody response to more than one antigen is indicative of a lower frequency of malarial infection. This gives scientific credibility to the notion that a successful malaria vaccine should contain multiple antigens. PMID- 15541031 TI - Enhanced inducible nitric oxide synthase expression and nitrotyrosine accumulation in experimental granulomatous hepatitis caused by Toxocara canis in mice. AB - The involvement of inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) in pathogenesis of toxocaral granulomatous hepatitis (TGH) in a murine host was quantitatively determined by biochemical, parasitological, pathological, and immunohistochemical assessments in a 42-week investigation. Mice were sacrificed for serum collection and histological processing as well as acid-pepsin digestion of the liver in a larval recovery study. Significantly increased levels of total serum NO were found in the trial, indirectly suggesting iNOS activation in the liver. iNOS reactivity was predominantly observed in infiltrating leucocytes in lesions and normal and apocrine-like cholangiocytes; in contrast, hepatocytes and multinucleated giant cells showed negative cytoplasmic staining in TGH. Strong iNOS-like reactivity was also detected on the body wall of larvae. The locations of NT reactivity were nearly identical to those of iNOS expression; infiltrating leucocytes or cholangiocytes stained for iNOS were also stained for NT in TGH. Enhanced iNOS expression, but not invading larvae (r = 0.256, P = 0.211), seemed to play a certain role in pathological damage in TGH due to a significant correlation between iNOS expression and serum alanine aminotransferase (ALT) levels (r =0.593, P = 0.021) in the trial. Our present results indicate a potential therapeutic strategy for treatment of GH caused by other nematodes through manipulation of iNOS expression. PMID- 15541032 TI - Adjuvant guided polarization of the immune humoral response against a protective multicomponent antigenic protein (Q) from Leishmania infantum. A CpG + Q mix protects Balb/c mice from infection. AB - It has been shown that vaccination with three doses of the Leishmania infantum poly-protein Q containing five genetically fused antigenic determinants from the Lip2a, Lip2b, H2A and P0 proteins, mixed with BCG induces clearance of parasites in 9 out of 10 Leishmania infantum-infected Beagle dogs, in addition to clinical protection. In the present paper we analysed the immunogenic potential of the poly-protein Q and the specificity and polarization of the response against the antigenic determinants of Q when mixed with various adjuvants. The data showed that the Q protein had high intrinsic immunogenic potential and that it was able to induce a long-lasting IgG response. The IgM immunogenic potential of the poly protein was mainly due to the LiP2a and LiP2b determinants, whereas the IgG immunogenic potential was mainly due to the LiP2a component. It was observed that the protein itself elicited a mixed IgG2a/IgG1 response and that the determinants of Q were endowed with different IgG2a/IgG1 potential. It was also observed that the adjuvants did not influence the intensity or specificity of the IgM response but that they modulated the intensity, the specificity and the polarization of the IgG response against the determinants of Q. CpG-ODN motifs or double-stranded DNA plasmids containing CpG motifs when mixed with Q induced a predominant IgG2a response mainly observed at early stages post-immunization. The data showed that a CpG + Q mix induced significant protection against L. infantum infection in Balb/c mice. PMID- 15541033 TI - Saliva of the Yellow Fever mosquito, Aedes aegypti, modulates murine lymphocyte function. AB - Saliva of many vector arthropods contains factors that inhibit haemostatic responses in their vertebrate hosts. Less is known about the effect of vector saliva on host immune responses. We investigated the effect of Aedes aegypti salivary gland extracts on antigen-stimulated responses of transgenic OVA-TCR DO11 mouse splenocytes in vitro. T-cell proliferation was inhibited in a dose dependent manner, with greater than 50% inhibition at 0.3 salivary gland pair (SGP) equivalents/mL. LPS-stimulated B-cell proliferation was also inhibited. Secretion of the Th1 cytokines IL-2 and IFN-gamma was reduced by 50% or more with 0.45-0.6 SGP/mL, as was secretion of the pro-inflammatory cytokines GM-CSF and TNF-alpha, and the Th2 cytokine IL-5. The Th2 cytokines IL-4 and IL-10 were similarly reduced with 0.6-2 SGP/mL. Inhibition of lymphocyte function involved modulation of viable T-cells at low salivary gland extract (SGE) concentrations, and decreased viability at higher concentrations. Dendritic cells were not killed by salivary gland extracts at concentrations as high as 25 salivary gland pairs/mL, but secretion of IL-12 was inhibited by 87% following exposure to 0.6 SGP/mL. Activity is present in saliva and extracts of female but not male salivary glands, and it is depleted from salivary glands of blood-fed mosquitoes. The activity is denatured by boiling and by digestion with the protease papain, indicating a protein; gel filtration HPLC indicates a mass of about 387 kDa. These results suggest that A. aegypti saliva exerts a marked immunomodulatory influence on the environment at the bite site. PMID- 15541034 TI - Immunization against sheep scab: preliminary identification of fractions of Psoroptes ovis which confer protective effects. AB - In an attempt to enrich for potentially protective Psoroptes ovis antigens, three separate vaccine trials were conducted in which groups of sheep were immunized three times with various fractions of a soluble extract of P. ovis mites using QuilA as adjuvant. These groups, as well as controls that received adjuvant only, were challenged with P. ovis, and protective immunity was assessed by measuring lesion areas and conducting mite counts 4 and 6 weeks later. All fractions stimulated high titre serum antibodies. Most conferred some protection on sheep with active disease, although there was considerable variation between sheep in all groups, including the controls. Some fractions were more protective than the extract itself, suggesting that the protective components had been concentrated. Indeed the best fraction, obtained by ion exchange chromatography, followed by a gel filtration step, slowed lesion growth to less than a third by 6 weeks after challenge and reduced mite numbers by more than 13 times compared to control sheep vaccinated with QuilA only. However, as judged by polyacrylamide gels, the polypeptide profile of this fraction was still complex, indicating that further work is required to identify the protective components. PMID- 15541035 TI - Interferon-gamma mediates neuronal killing of intracellular bacteria. AB - Neurons can be targets for microbes, which could kill the neurons. Just in reverse, we, in this study, report that bacteria can be killed when entering a neuron. Primary cultures of foetal mouse hippocampal neurons and a neuronal cell line derived from mouse hypothalamus were infected by Listeria monocytogenes. Treatment with interferon-gamma (IFN-gamma) did not affect bacterial uptake, but resulted in increased killing of intracellular bacteria, whereas the neuronal cell remained intact. The IFN-gamma-mediated bacterial killing was mapped to the neuronal cytosol, before listerial actin tail formation. Treatment with IFN-gamma induced phosphorylation of the transcription factor STAT-1 in neurons and IFN gamma-mediated listerial killing was not observed in STAT-1(-/-) neurons or neurons treated with IFN regulatory factor-1 antisense oligonucleotides. IFN gamma-treated neuronal cells showed increased levels of inducible nitric oxide synthase (iNOS) mRNA, and antisense iNOS oligonucleotides hampered the bacterial killing by neurons upon IFN-gamma treatment. This novel neuronal function - i.e., that of a microbe killer - could play a crucial role in the control of infections in the immuno-privileged nervous system. PMID- 15541036 TI - Expression of interleukin-22 in murine carcinoma cells did not influence tumour growth in vivo but did improve survival of the inoculated hosts. AB - Interleukin (IL)-22, a novel cytokine belonging to the IL-10 family, is secreted from activated T and natural killer cells and is possibly involved in inflammatory responses. We examined whether expression of the IL-22 gene in murine colon carcinoma Colon 26 cells (Colon 26/IL-22) could produce any antitumour effects in the inoculated mice. Although growth of Colon 26/IL-22 tumours in syngeneic mice was not different from that of parent tumours, survival of the mice that were subcutaneously or intraperitoneally inoculated with Colon 26/IL-22 tumours was significantly prolonged compared with the mice inoculated with parent tumours. Metastasis was not influenced by IL-22 expressed in tumours. Expression of the IL-22 receptor-specific gene, IL-22R, was not induced in spleen cells stimulated with concanavalin A, anti-CD3 or anti-CD40 antibody, despite constitutive expression of the IL-10R2 gene, which encodes another component of the heterodimeric IL-22 receptor complex. IL-22 thereby does not directly act on immunocompetent cells, and IL-22 expressed in tumours can favour apothanasia of inoculated hosts. PMID- 15541037 TI - Platelet-derived growth factor-AA increases IL-1beta and IL-8 expression and activates NF-kappaB in rheumatoid fibroblast-like synoviocytes. AB - The effect of platelet-derived growth factor (PDGF)-AA on the inflammation in rheumatoid arthritis (RA) and osteoarthritis (OA) was investigated using cultured fibroblast-like synoviocytes (FLS) obtained from RA and OA patients as well as control nonarthritic (NA) individuals. PDGF-AA increased the mRNA and protein expressions of proinflammatory cytokines, interleukin (IL)-1beta and IL-8 in RA FLS. Biological activity of IL-1 in the culture supernatant of RA FLS was also increased by PDGF-AA stimulation. Interestingly, PDGF-AA synergized with tumour necrosis factor (TNF)-alpha to upregulate the protein expressions of IL-1beta and IL-8. PDGF-induced enhancement of the IL-1beta and IL-8 mRNA expressions was also observed in OA FLS. However, the expression of these proinflammatory cytokines in NA FLS did not change by PDGF treatment, suggesting that the inflammatory condition might have modified the biological effects of PDGF. In accordance with the enhanced expression of inflammatory cytokines, the activity of nuclear factor kappaB was also induced in response to PDGF-AA in RA FLS. These results suggest that PDGF-AA plays an important role in the progression of RA inflammation, and inhibiting PDGF activity may be useful for the effective RA treatment. PMID- 15541038 TI - High-polarity Mycobacterium avium-derived lipids interact with murine macrophage lipid rafts. AB - Cholesterol- and sphingolipid-rich membrane microdomains (lipid rafts) are widely recognized as portals for pathogenic micro-organisms. A growing body of evidence demonstrates mobilization of host plasma cell membrane lipid rafts towards the site of contact with several pathogens as well as a strict dependence on cholesterol for appropriate internalization. The fate of lipid rafts once the pathogen has been internalized and the nature of the pathogen components that interact with them is however less understood. To address both these issues, infection of the J774 murine cell line with Mycobacterium avium was used as a model. After demonstrating that M. avium induces lipid raft mobilization and that M. avium infects J774 by a cholesterol-dependent mechanism, it is shown here that mycobacterial phagosomes harbour lipid rafts, which are, at least in part, of plasma cell membrane origin. On the other hand, by using latex microbeads coated with any of the three fractions of M. avium-derived lipids of different polarity, we provide evidence that high-polarity, in contrast to low-polarity and intermediate-polarity, mycobacterial lipids or uncoated latex beads have a strong capacity to induce lipid raft mobilization. These results suggest that high polarity mycobacterial lipid(s) interact with host cell cholesterol-enriched microdomains which may in turn influence the course of infection. PMID- 15541039 TI - Host-dependent serum specificity to the V3 domain of HIV-1. AB - The previous studies of anti-HIV-1 humoral immune response have found the stable individual structural bias in the antibody production that reflects the changes in the immune functional network caused by HIV-1. The correlation between antigen structure, in particular the V3 domain of HIV-1, and the serum specificity to the structure remains unclear. To clarify the role of host individual factors in the serum specificity, we used the slightly modified HIV-1 serotyping methodological approach for the sera of the patients infected with HIV-1 with a different level of genetic similarity. Substantial discrepancy between serum specificities and antigen structure was found. Patients infected with HIV-1 carrying similar and identical V3 sequences had significantly different serum specificities. The opposite situation was often observed as well. The results of the study suggest that the influence of the V3 loop structure in the development of humoral immune response to the V3 loop is substantially modified in a patient-specific manner. PMID- 15541040 TI - Modulation of cytokine release by differentiated CACO-2 cells in a compartmentalized coculture model with mononuclear leucocytes and nonpathogenic bacteria. AB - To further investigate the interaction between human mononuclear leucocytes [peripheral blood mononuclear cells (PBMC)] and enterocytes, the effect of a confluent layer of differentiated CACO-2 cells on cytokine kinetics during challenge with bacteria in a compartmentalized coculture model was investigated. Nonpathogenic Escherichia coli were added either to the apical or the basolateral compartment of this transwell cell culture system, the latter of which contained human leucocytes. The synthesis of tumour necrosis factor (TNF-alpha) and interleukin (IL)-12 was significantly suppressed by CACO-2 cells when leucocytes were stimulated directly with bacteria. This suppression was not paralleled by changes in the production of IL-10, IL-6 and transforming growth factor (TGF) beta. When the bacteria were applied apically to the CACO-2 cell layer, the production of TNF-alpha, IL-12, IL-1beta, IL-8, IL-6, IL-10, TGF-beta and interferon-gamma was pronouncedly lower as compared to the bacterial stimulation of leucocytes beneath the CACO-2 cells. In the latter experiments, IL-6, IL-8 and TNF-alpha were the cytokines being mostly induced by apical addition of E. coli. Quantitative mRNA expression analysis revealed that IL-8 gene expression was equally induced in both CACO-2 and PBMC after apical stimulation with bacteria. Of note, bacteria-stimulated CACO-2 cells produced little or no cytokines in the absence of leucocytes, supporting the concept of leucocyte-epithelial cell cross talk in modulating cytokine responses in the gut mucosa. PMID- 15541041 TI - Evaluation of the allergenicity and antigenicity of bovine-milk alphas1-casein using extensively purified synthetic peptides. AB - Alphas1-Casein (CAS1_BOVIN), the major allergen of cow's milk (CM), is widely used as hydrolysates in infant diet formulae and additive to other processed food items. To date, most of the reported B-cell epitope mapping were performed on polyethylene pins or cellulose-derivative membrane. We sought to locate the motifs critical for human-specific IgE and rabbit polyclonal IgG binding using extensively purified CAS1_BOVIN, synthetic peptides and derivatives. Thirteen overlapping peptides covering the whole CAS1_BOVIN encompassing 17 : 20 amino acid (AA) were synthesized by f-moc AA solid-phase polyamide peptide synthesis. In addition, six cyanogen bromide (CNBr) cleavage fragments were prepared. Limited hydrolysis, oxidized and reduced/alkylated derivatives were also produced. The preparations were purified by ion exchange, gel filtration chromatography, reversed phase and high-performance liquid chromatography. The homogeneity was visualized by sodium dodecyl sulfate (SDS) and poly acryl amide gel electrophoresis (PAGE) followed by IgE and IgG immunoblotting. IgE binding was measured by Biotin Streptavidin (Bio/strep) fluoro enzyme immuno assay (FEIA) or ELISA-inhibition. Eighteen CM allergy (CMA) sera from 45 clinically examined children (Melbourne) and five adults (Bergen) were selected. Individual sera and pools were used for mapping IgE-binding epitopes. Rabbit IgG sera and pools were used for locating the antigenic sites of the molecule. Results indicated that all the individual CMA sera and pools recognized the intact molecule and three of the CNBr fragments as major antibody-binding allergens. The N- and C-terminal peptides (CAS 16-35; CAS 136-155) showed high IgE-binding affinity. CAS 1-18 and CAS 181-199 showed high IgG bindings. Considering the diversity of the antibody specificities, a reasonable agreement between IgE and IgG epitopes were found at the N- and C-terminals of CAS1_BOVIN. Mapping IgE B-cell epitopes by direct Bio/strep FEIA allowed the development of a sensitive modified technique for detecting unlabelled, casein immune dominant peptides in food products. PMID- 15541042 TI - Differential expression of the C5a receptor and complement receptors 1 and 3 after LPS stimulation of neutrophils and monocytes. AB - Animal experiments recently suggested that administration of anti-C5a, anti-C5a receptor or soluble complement receptor type-1 may be of value in the treatment of septic shock. Because results regarding C5a receptor expression (C5a-R, CD-88) have been found to differ between septic animals and patients, the aim of this study was to investigate the neutrophil and monocyte receptor expression of CD-88 and complement receptor-1 (CR-1, CD-35) after stimulation with lipopolysaccharide (LPS) ex vivo. Whole blood or isolated neutrophils and monocytes from healthy people were incubated with LPS in a dose range of 0.1-1000 ng/ml. The expressions of CD-88 and CD-35 were analysed by means of flow cytometry. For comparison, the expressions of complement receptor-3 (CR-3, CD-11b/CD-18), Fc-gamma receptor type I (CD-64) and CEACAM-8 (CD-66b) were also investigated. In whole blood, CD-88 expression on neutrophils was reduced (P < 0.05). The expressions of CD-35 and CD 11b were increased both on neutrophils (P < 0.001; P < 0.05) and on monocytes (P < 0.001; P < 0.001). No effect was observed on isolated cells. In agreement with the findings in septic patients, LPS reduced the neutrophil C5a-R expression, whereas the expressions of CR-1 and CR-3 were increased. The effects of LPS were indirect and were mediated via factors in the blood. The clinical significance of this is not known, but may be associated with decreased chemotaxis. PMID- 15541043 TI - Modulation of CD28 and CD86 expression in patients with paracoccidioidomycosis in different periods of treatment. AB - Paracoccidioides brasiliensis, a thermo-dimorphic fungus, is the ethiologic agent of paracoccidioidomycosis (PCM). The recidive is the greatest obstacle of this disease, because the yeast usually returns after the long treatment period. In the present work, we have investigated the cellular immune response of cells from peripheral blood drawn from patients with different duration of PCM. The classification of patients ranged from nontreated to those with long-standing disease over 5 years. Unstimulated as well as cells stimulated with phytohemaglutinin or two different antigen preparations, secreted (MEXO) or somatic (PbAg) of P. brasiliensis, were characterized. We found that cells from patients with disease proliferate considerably upon stimulation with the antigen preparations and that cells from patients with disease of long duration does not proliferate that vigorously as from patients with more recent diagnosis. Both interferon (IFN)-gamma and interleukin (IL)-4 appear to be increased in patients, but IFN-gamma tended to increase upon treatment while IL-4-secretion decreased. With respect to CD28 and CD86, we found that the subset of CD28 positive CD8 cells are decreased in all stages of the disease as compared to control individuals. A subset of CD86 positive CD19 cells appeared to be considerably increased compared to the controls. Indeed, our results demonstrated that the treatment of PCM patients promoted a regulation of IFN-gamma, IL-4 levels and CD28, CD86 expression bringing new insight to the cellular immune response in PCM. PMID- 15541044 TI - Elicitation of both CD4 and CD8 T-cell-mediated specific immune responses to HCA587 protein by autologous dendritic cells. AB - We recently cloned a new member of cancer/testis antigen named HCA587, which was highly expressed in human hepatocellular carcinoma (HCC) tissues. To investigate it as a potential tumour-specific target for immunotherapy, the immunogenicity of this protein, especially the ability to induce specific cellular immune responses, was evaluated in the present study. As dendritic cells (DC) are the most potent antigen-presenting cells, DC-based vaccination has recently shown marked promise for the treatment of human malignancies by immunological intervention. Here, we demonstrate that autologous DC loaded with HCA587 protein could induce specific T-cell responses in healthy individuals by in vitro stimulations. Enzyme-linked immunospot analysis for interferon-gamma (IFN-gamma) secretion demonstrated HCA587-specific CD8(+) T cells in the antigen-stimulated peripheral blood lymphocytes, and the analysis of CD4(+) T cells by proliferation assay also showed antigen-specific reactivities in normal donors. Two-colour flow cytometric analysis of surface markers and intracellular cytokine expression demonstrated that HCA587-specific cytotoxic T lymphocytes exhibited a heterogeneous CD8(+)/CD56(+) expression, and a striking T-helper 1 cytokine bias (IFN-gamma(high)/IL-4(low)) was observed for both CD4(+) and CD8(+) HCA587 specific lymphocyte populations. We conclude that HCA587 is a potent immunogen that can induce CD4(+) and CD8(+) T-cell-mediated specific immune responses, and these findings propose HCA587 as a good candidate for the development of a therapeutic protein-based DC tumour vaccine for the treatment of HCC patients. PMID- 15541045 TI - Differential adherence of osteoarthritis and rheumatoid arthritis synovial fibroblasts to cartilage and bone matrix proteins and its implication for osteoarthritis pathogenesis. AB - In osteoarthritis (OA), cartilage and bone fragments have been described within the synovial tissue which are surrounded by synovial cells (i.e. detritus synovitis). These cells appear to attach actively to the cartilage and bone fragments. In rheumatoid arthritis (RA), on the other hand, synovial fibroblasts (SF) have also been shown to be localized at sites of invasion into cartilage and bone and to degrade extracellular matrix (ECM) by secreting proteolytic enzymes. One prerequisite for exerting their aggressive properties is the attachment to cartilage and bone ECM. This attachment appears to be mediated by the expression of different adhesion molecules for which corresponding binding sites on ECM components are known. As it has not been addressed to which ECM proteins SF adhere and with which affinity this process takes place, we investigated the adherence of SF from patients with OA and RA to different cartilage and bone matrix proteins. Synovial tissue samples were obtained during synovectomy or arthroplastic surgery and used for isolating and culturing SF. Synovial cells attaching to cartilage/bone fragments were characterized using immunohistochemistry. The adherence of SF to ECM proteins was examined using an adhesion assay with the following proteins coated on 96-well plates: aggrecan (AGG), bone sialoprotein (BSP), cartilage oligomeric matrix protein (COMP), collagen type I, II and VI, proline arginine-rich, end leucine-rich repeat protein (PRELP), osteopontin (OPN) and recombinant chondroadherin (CHAD). Bovine serum albumin was used as negative control. In addition, adhering fibroblasts were photographed using a phase-contrast microscope. As compared with RA-SF, significantly higher numbers of OA-SF adhering to collagen type II, OPN and CHAD could be detected (P < 0.05). In contrast, RA-SF showed increased attachment to collagen type II, OPN and BSP. Adhesion to AGG, COMP and PRELP appeared not to be significantly increased and differed widely among the SF samples, and, apart from one exception (BSP), OA-SF adhered in higher numbers to the matrix proteins than did RA-SF. Using immunohistochemistry, synovial cells attached to cartilage/bone fragments could be shown to predominantly express CD68 (>/=50%). The CD68 negative population was of the fibroblast phenotype (AS02 positive). The study demonstrates that the binding pattern of OA-SF and RA-SF to ECM proteins differs considerably and therefore provides novel insights into the difficult pathophysiology of OA and RA. In general, it appeared that SF adhere primarily to ECM proteins that contain known binding sites for adhesion molecules (e.g. integrins: collagen/integrin alpha(2)beta(1)) and that higher numbers of OA-SF adhered to the cartilage and bone matrix proteins than did RA-SF. PMID- 15541046 TI - Distribution of Entamoeba histolytica Gal/GalNAc lectin-specific antibody response in an endemic area. AB - To demonstrate the dynamics of specific antibody isotypes against Entamoeba histolytica Gal/GalNAc adhesin and its correlation, if any, with the development of immunity, we evaluated subjects suffering from a spectrum of amoebic infections ranging from amoebic liver abscess (ALA) to asymptomatic cyst passers. The quantitative analysis of antibody levels was done in the sera by enzyme linked immunosorbent assay. Gal/GalNAc adhesin-specific immunoglobulin G (IgG) was higher in ALA (and their follow-ups) and intestinal amoebiasis cases as compared with asymptomatic cyst passers (P < 0.05). Among the isotypes of IgG, high levels of IgG1 (60% of the total IgG concentration), suggestive of T-helper 2-type response, was associated with ALA cases. Intestinal amoebiasis cases and cyst passers had high percentage of IgG1 and IgG4 antibodies as compared with per cent IgG2 and IgG3 (of the total IgG), whereas follow-up cases of ALA had predominantly IgG2 and IgG3 isotypes of antibodies. Gal/GalNAc lectin-specific IgM antibodies were maximum in cases of intestinal amoebiasis. ALA cases and their follow-ups had significantly lower levels of Gal/GalNAc-specific IgM levels as compared with cyst passers (P < 0.05). Gal/GalNAc adhesin-specific IgA antibodies were raised maximally in intestinal amoebic infection cases. ALA cases and their 3-month follow-ups had significantly higher concentrations of lectin specific IgA (P < 0.05) as compared with the healthy subjects. PMID- 15541047 TI - Unique TCRDV1-positive lymphocytes that infiltrate colonic neoplasia originate from a selected cell population present in the intestinal mucosa and the peripheral blood. AB - TCRDV1-positive lymphocytes, which infiltrate colon carcinomas, were recently shown to be cytolytic for tumour cells. However, the immune compartment from which these cells originate is unknown. The aim of the present studies was to determine the origin of TCRDV1-positive cells in colonic neoplasia. Biopsies were obtained from normal colon, polyps or carcinomas, concurrently with a sample from the peripheral blood. RNA was extracted and a TCRDV1-specific reverse transcriptase-polymerase chain reaction (RT-PCR) was performed. Amplification products were analysed by a CDR3 display and sequence analysis. In five out of six patients, the TCRDV1 CDR3 display of the whole cell population within the neoplastic tissue was distinct from that in the normal mucosa and the peripheral blood. The nucleotide sequences of CDR3 domains from the three compartments were distinct as well. In one patient, a pattern similar to the CDR3 display was detected in neoplastic and normal intestinal tissues. However, using junction specific RT-PCR of CDR3 sequences derived from the neoplastic cells, such sequences could be detected in all three compartments. These findings suggest that in contrast to the current paradigm, a unique TCRDV1 cell population circulates in the peripheral blood and normal intestinal tissue and infiltrates colon neoplasia rather than being restricted to a single compartment as previously thought. PMID- 15541048 TI - Release of TNF-alpha from in vitro-stimulated monocytes is negatively associated with serum levels of apolipoprotein B in patients with type 2 diabetes. AB - Impaired course of inflammation is a likely mechanism behind a number of diabetic complications. The present study was undertaken to investigate lipopolysaccharide induced production of tumour necrosis factor (TNF)-alpha in monocytes from patients with type 2 diabetes and to assess its relationship with diabetes associated metabolic abnormalities. Monocytic TNF-alpha mRNA production was lower in the diabetic participants compared to their corresponding controls. Diabetic subjects who had been receiving simvastatin treatment had TNF-alpha mRNA production similar to that of the healthy participants. The release of TNF-alpha from diabetic cells correlated negatively with serum levels of apolipoprotein B (apoB) (R = -0.755, P = 0.001), total plasma cholesterol (R = - 0.702, P = 0.002) and the presence of retinopathy (R = -0.572, P = 0.021). No such associations were found in the control subjects. In a multiple linear regression model, only the level of apoB and diabetes duration demonstrated significant effects on the release of TNF-alpha, with apoB alone accounting for 57% of the variation. We conclude that production of TNF-alpha mRNA in response to the bacterial stimulant is compromised in poorly controlled type 2 diabetes. Lipid abnormalities are associated with the observed defect. Impaired cytokine production represents a significant defect in the functioning of the immune system and may contribute to aberrations in the course of inflammation in the diabetic state. PMID- 15541049 TI - The male reproductive system and its susceptibility to endocrine disrupting chemicals. AB - In the past years, there has been increased interest in assessing the relationship between impaired male fertility and environmental factors. Human male fertility is a complex process and therefore a great variety of sites may be affected by exogenous noxae. Lifestyle factors as well as various environmental and occupational agents may impair male fertility. Many studies have been published reporting on reproductive dysfunctions in male animals and humans. Especially environmental pollutants with endocrine activity are discussed as a possible cause of this detrimental development. Evidence from animal experiments show that substances with oestrogenic and antiandrogenic properties may cause hypospadia, cryptorchidism, reduction of sperm density and an increase of testicular tumours. Many adverse effects on animal male fertility have been documented for phthalates and some chlorinated hydrocarbons such as polychlorinated biphenyls and polychlorinated dibenzo-p-dioxins. For other chemicals such as bisphenol A and nonylphenols animal data are conflicting. Environmental pollutants may mediate their effects by receptor binding, modulation of hormone-regulated mechanisms or direct toxic effects. Data on environmental chemicals and human male fertility are scarce, and risk assessment is mostly based on the results of animal studies. However, there are indications that exposure to endocrine active chemicals during early development may alter hormone responsiveness in adulthood. Furthermore, some of the chemicals are found in fluids that are associated with human reproduction, such as follicular fluid, seminal fluid and cervical mucus. Recent studies suggest a correlation between pesticide exposure and standard semen parameters as well as in vitro fertilization rates. PMID- 15541050 TI - Male breast cancer--an andrological disease: risk factors and diagnosis. AB - Gynaecomastia, the enlargement of the male breast, is considered as an andrological disease. To date, a review on male breast cancer (MBC) has not been published in an andrological journal. The papers underlying this review were published from authors of different institutions: Clinical Genetics, Dermatology, Gynaecology, Internal Medicine, Oncology, Pathology, Psychiatry, Radiology and Surgery. MBC accounts for approximately 1% of breast cancer patients. A total of 182 men died of breast cancer in 1999, in Germany. In the US, 1500 new cases per year occur. MBC accounts for <5% of surgically removed breast lumps. Diseases with increased oestrogen action increase the risk of MBC. Mutations of distinct genes are estimated to account for up to roughly 10% of MBC. BRCA1 and BRCA2 gene mutations are responsible for approximately 80% of the families with hereditary breast cancer. The diagnosis of MBC is not possible without histological examination. Different diagnostic procedures such as clinical diagnosis, sonography, mammography, fine-needle biopsy and core needle facilitate the decision whether a biopsy is necessary. PMID- 15541051 TI - Testosterone therapy in the ageing male: what about the prostate? AB - The concerns about testosterone therapy in ageing men with late-onset hypogonadism mainly address the risk of prostatic disease, i.e. either benign prostatic hyperplasia (BPH) or prostate cancer (PCa). Both conditions are highly dependent on androgen action and recent clinical data on the cancer-preventive effect of the 5alpha-reductase inhibitor finasteride have supported the possible role of androgens in PCa. However, the clinical data especially on the long-term effects of exogenous androgen substitution in regard to prostate safety are nonconclusive in many respects. As sufficient clinical studies on these risks will not be available in the near future, the approach of testosterone therapy towards prostate complications should be kept on a safe but practical basis. This review includes some recommendations in regard to testosterone therapy and prostate monitoring in patients with BPH and bladder outlet obstruction, with previous history of curative treatment for PCa or with prostatic intraepithelial neoplasia. PMID- 15541052 TI - Use of a laser to detect viable but immotile spermatozoa. AB - A prerequisite for the success of intracytoplasmic sperm injection (ICSI) is the use of viable sperm that will support oocyte activation and embryo development. Usually, spermatozoa are selected according to motility, which is a clear indication for viability. However, in cases with complete asthenozoospermia, it is impossible to select viable sperm by morphological means. Here we report for the first time on the use of a laser to discriminate between viable and dead immotile spermatozoa. The application of a single laser shot to the far end of the sperm tail causes a curling of the sperm tail only in viable sperm, similar to the reaction observed in the hypo-osmotic swelling test. Using this method for sperm selection prior to ICSI we achieved high fertilization and cleavage rates in cases with immotile sperm in fresh testicular biopsy material as well in cases with ejaculated immotile sperm. It is concluded that laser sperm interaction allows for the identification of viable but immotile spermatozoa. PMID- 15541053 TI - Correlation between tyrosine phosphorylation intensity of a 107 kDa protein band and A23187-induced acrosome reaction in human spermatozoa. AB - This study, performed using semen samples from 10 men, investigated the relationship between sperm protein tyrosine phosphorylation and acrosomal status in conditions supporting in vitro capacitation. Percoll-selected spermatozoa (cells from the 95% fraction) were incubated for 3 h at 37 degrees C under an atmosphere of 5% CO2 in air, in a polyvinyl alcohol (1 mg ml(-1)) containing Biggers-Whitten-Whittingham's medium, nonsupplemented or supplemented with either bovine serum albumin (BSA; fatty acid free, 3 mg ml(-1)) or 2-hydroxy-propyl-beta cyclodextrin (2-OH-p-beta-CD; 0.5, 1, 2 mmol l(-1)). Sperm suspension in each medium was split into two aliquots. The first was used to evaluate the acrosomal status by staining with the fluorescein isothiocyanate Pisum sativum agglutinin after induction of the acrosome reaction (AR) for 45 min with 10 micromol l(-1) of A23187 calcium ionophore. The second aliquot was used for sodium dodecyl sulphate polyacrylamide gel electrophoresis and immunoblotting, followed by a densitometric analysis. Compared with the nonsupplemented medium, BSA- or 2-OH-p beta-CD-supplementation induced an increase in both the percentage of live acrosome-reacted sperm and the tyrosine phosphorylation intensity of the main phosphorylated 107 kDa protein. A correlation between the percentage of live acrosome-reacted sperm and the 107-kDa protein phosphotyrosine intensity was observed. Therefore, the 107 kDa protein-phosphotyrosine level measurement would bring additional information to conventional semen parameters in the assessment of the human sperm functionality. PMID- 15541054 TI - Overactive corpus cavernosum: a novel cause of erectile dysfunction. AB - Our recording of the electromyographic (EMG) activity of the corpus cavernosum (CC) in 59 patients with erectile dysfunction (ED) revealed 18 patients who had elevated electric activity, which presumably points to heightened tone of the CC smooth muscles. We investigated the hypothesis that this elevated EMG activity and muscular tone of the CC could be the cause of ED. The study comprised the said 18 subjects with the hypertonic CC muscles as study group (42.6 +/- 5.3 SD years), 15 healthy volunteers (41.8 +/- 5.1 SD years) and 15 patients (41.6 +/- 5.5 SD years) with ED who had not recorded elevated tone of the CC muscles as control group. The EMG activity was registered in the flaccid, erectile and detumescent phases by two electrodes inserted into the CC. Electrocavernosography (ECG) of healthy volunteers recorded in the flaccid phase showed regular slow waves (SW) and random action potentials (APs). The wave variables declined significantly in the erection phase (P < 0.01). In the study group, the SW variables in the flaccid phase increased significantly (P < 0.05) compared with the healthy volunteers and the rhythm was irregular. Erection did not occur with sildenafil but with intracavernosal injection of papaverine, which led to decline of the SW variables (P < 0.05). The control ED group exhibited in the flaccid phase diminished SW variables (P < 0.05) compared with the healthy volunteers. On erection with sildenafil administration, the SW variables showed significant reduction (P < 0.05). CC hypertonicity or 'overactive CC' was identified as a possible cause of ED. An elevated EMG activity of the CC muscle fibres in the flaccid phase presumably denotes hypertonicity of these fibres and their failure to relax to effect erection. The cause of elevated CCEMG activity and presumed muscle hypertonicity is unknown and could be functional or organic. Erection was produced by intracavernosus injection of papaverine and not by sildenafil. This condition of 'overactive CC' should be considered in the diagnosis of ED. However, further studies in the pathogenesis of the condition are warranted. PMID- 15541055 TI - Effect of paternal heat stress on the development in vitro of preimplantation embryos in the mouse. AB - This study was conducted to investigate the effects of paternal heat stress on the development in vitro of preimplantation embryos in the mouse. Female C57/CBA mice were superovulated using eCG/hCG and mated either to an untreated (control) male mouse or to one that had been exposed for 24 h to an ambient temperature of 36 +/- 0.1 degrees C and 62 +/- 0.4% relative humidity, between 3 and 42 days previously. Putative zygotes were collected from the oviducts of mated mice, 25 28 h after hCG injection, and cultured in vitro. Embryo development was evaluated at 24-h intervals for up to 120 h. Paternal heat stress significantly reduced the proportion of embryos that developed normally during 24-120 h of in vitro culture, when zygotes were sired by males which had been heat stressed between 7 and 35 days prior to mating. Maximum impairment to development (including nondevelopment, abnormal and dying/dead embryos) occurred in those embryos sired by males at days 14 and 21 after heating. Embryo development returned to control levels by day 42 after heat stress. Furthermore, whilst all stages of embryo development were affected by paternal heat stress, the proportion of embryos at the two-cell stage appeared to be most severely affected. Four-cell to morula stages and the morula to blastocyst stage also demonstrated impairment at days 14, 21, 28 and 35 after heating. These results demonstrate that a single episode of paternal heat stress significantly reduces the development of preimplantation embryos, and this is not recovered until day 42 after heating. The present results also support previous work demonstrating that sperm from the epididymis as well as germ cells in the testis are susceptible to damage by heat stress, with both spermatids and spermatocytes being the most vulnerable. PMID- 15541056 TI - Modulation of human sperm function by follicular fluid. AB - Human follicular fluid (hFF), present in the ampullary environment, can reduce the number of sperm bound to the zona pellucida. The aim of the present study was to investigate the effects of follicular fluid on sperm function. The presence of 50% v/v follicular fluid resulted in a significant reduction in the number of bound spermatozoa with respect to control medium (12.7 +/- 5.5 sp HZ(-1) versus 24.6 +/- 5.7 sp HZ(-1), P = 0.03) as measured by the hemizona binding assay. This reduction in zona binding capacity was not associated with a loss of sperm viability, motility or a premature acrosomal reaction. When capacitated spermatozoa were previously exposed 1 h to follicular fluid, a significant reduction in the number of alpha-d-mannose binding sites on sperm head was detected (23.7 +/- 3.1% versus 15.5 +/- 2.4%, P < 0.05). In addition, sperm fertilizing capacity (assessed as the acrosome reaction to ionophore challenge score) in the presence of follicular fluid was also diminished (38.0 +/- 4.8% versus 22.6 +/- 4.9%, P < 0.01). No modification in the pattern of protein tyrosine phosphorylation which occurs during capacitation was observed in the presence of the fluid. Taken together, the results indicate that the decrease in sperm zona-binding capacity observed in the presence of hFF was related to a lower number of sperm containing alpha-d-mannose receptors. PMID- 15541057 TI - Which is the association between erectile dysfunction and lower urinary tract symptoms? PMID- 15541060 TI - Clinical nurse specialist care managers' time commitments in a disease-management program for bipolar disorder. AB - OBJECTIVES: As part of a cost-effectiveness analysis for Department of Veterans Affairs Cooperative Studies Program #430, 'Reducing the Efficacy-Effectiveness Gap in Bipolar Disorder,' we conducted a time and motion study to quantify the time psychiatric clinical nurse specialist (CNS) care managers spent providing care for patients. METHODS: Clinical nurse specialist care managers completed activity logs in which they recorded time spent implementing the Bipolar Disorders Program (BDP) during a 1-week period in spring, summer, fall and winter over a 1-year period when caseloads were at steady state. Mean service time was estimated by use of univariate analysis of means and by multivariable regression analysis. RESULTS: On average CNS care managers spent 40% of their clinical time in activities that typically are reimbursed (e.g. clinic visits) and spent the remaining 60% of their time in activities that are typically unreimbursed. Total clinic time increased as the number of visits per day increased; however, this increase got smaller with each additional visit per day. CONCLUSIONS: As with other chronic illness management programs, CNS care managers expend a substantial portion of their clinical effort for the BDP in activities that are typically unreimbursed. Their activities have a fixed component per day as well as a component that systematically varies with the number of visits per day. These findings should be considered when costing out and disseminating psychiatric and other medical chronic illness management programs. PMID- 15541061 TI - Demographic and diagnostic characteristics of the first 1000 patients enrolled in the Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD). AB - OBJECTIVES: Bipolar disorder is a severe, recurrent, and often highly impairing psychiatric disorder. The Systematic Treatment Enhancement Program for Bipolar Disorder (STEP-BD) is a large-scale multicenter study funded by the National Institute of Mental Health (NIMH) to examine the longitudinal course of the disorder and the effectiveness of current treatments. The current report provides a context for interpreting studies resulting from STEP-BD by summarizing the baseline demographic and diagnostic characteristics of the first 1000 enrolled. METHODS: The majority of the sample met DSM-IV criteria for bipolar I disorder (71%). Mean age of patients was 40.6 (+/-12.7) years and mean duration of bipolar illness was 23.1 (+/-12.9) years. Among the first 1000 subjects enrolled, 58.6% are females and 92.6% Caucasian. This report compares the STEP-BD sample with other large cohorts of bipolar patients (treatment and community samples). RESULTS: Compared with US population and community studies, the first 1000 STEP BD patients were less racially diverse, more educated, had lower income, and a higher unemployment rate. Results are discussed in terms of the contributions of STEP-BD (and other large-scale treatment studies) in understanding the nature, treatments, and outcomes of bipolar disorder for patients seeking care at academic treatment centers. CONCLUSIONS: The current report provides a context for interpreting future studies resulting from STEP-BD. The comparison of demographic and clinical characteristics between the samples across clinic-based studies suggests broad similarities despite the substantial differences in geography, payer mix, and clinical entry point. PMID- 15541062 TI - Posttraumatic stress disorder in patients with bipolar disorder: a review of prevalence, correlates, and treatment strategies. AB - OBJECTIVES: In this article, we review the evidence for, and implications of, a high rate of comorbid posttraumatic stress disorder (PTSD) in individuals with bipolar disorder. METHODS: We reviewed studies providing comorbidity data on patients with bipolar disorder, and also examined the PTSD literature for risk factors and empirically supported treatment options for PTSD. RESULTS: Studies of bipolar patients have documented elevated rates of PTSD. Based on our review, representing 1214 bipolar patients, the mean prevalence of PTSD in bipolar patients is 16.0% (95% CI: 14-18%), a rate that is roughly double the lifetime prevalence for PTSD in the general population. Risk factors for PTSD that are also characteristic of bipolar samples include the presence of multiple axis I disorders, greater trauma exposure, elevated neuroticism and lower extraversion, and lower social support and socio-economic status. CONCLUSIONS: These findings are discussed in relation to the cost of PTSD symptoms to the course of bipolar disorder. Pharmacological and cognitive-behavioral treatment options are reviewed, with discussion of modifications to current cognitive-behavioral protocols for addressing PTSD in individuals at risk for mood episodes. PMID- 15541063 TI - A perspective on the use of psychoeducation, cognitive-behavioral therapy and interpersonal therapy for bipolar patients. AB - OBJECTIVES: Although pharmacological treatment is at present essential for treating bipolar patients, a number of psychological interventions have recently been shown to be efficacious as add-on therapies for the prophylactic treatment of bipolar illness. The study aimed critically to examine the efficacy of several tested patient-focused therapies. METHODS: A systematic review of the literature on this topic was performed, using MEDLINE, PSYCLIT and CURRENT CONTENTS. 'Bipolar', 'Psychotherapy', 'Psychoeducation', 'Interpersonal' and 'Cognitive behavioral' were entered as keywords. RESULTS: To date, psychoeducation and cognitive-behavioral therapy are the psychological interventions that have been shown to be more efficacious in the prophylaxis of new recurrences. There remains a need for studies investigating the role and efficacy of psychological interventions during acute phases of the illness. CONCLUSIONS: As their therapeutic goals are complementary, a combination of psychotherapy and pharmacotherapy may allow patients to achieve better symptomatic and functional recovery. Further research is needed to determine which patients may be better candidates for psychological interventions and to estimate the relative effects of the different components of psychological approaches on outcome. PMID- 15541064 TI - GAMIAN-Europe/BEAM survey II: cross-national analysis of unemployment, family history, treatment satisfaction and impact of the bipolar disorder on life style. AB - OBJECTIVES: In a previous report, we described the global analysis of the 'GAMIAN Europe/BEAM survey' carried out in order to gain a better understanding of what is like to live with a bipolar disorder (BD). We report here on a cross-national analysis of unemployment, family history, side effects, treatment satisfaction and the impact the disorder had and has on patients' perception of life style and quality of life. METHODS: The methodology has been described in the previous report [Morselli PL and Elgie R, J Bipolar Disord (2003) 5, 265]. The analysis was carried out on the data derived from eight countries (France, Italy, Holland, Portugal, Russia, Scotland, Spain and Sweden) on a total of 968 respondents. Data from three other countries were not evaluated because of the low number of respondents. RESULTS: The data suggest that the problems and difficulties encountered by bipolar patients are similar throughout the various European countries, regardless of the political, social or cultural settings. The disease leads to a very high rate of unemployment and has a significant negative impact on the perception of the quality of life, both within and outside the family. Data also indicate a high level of family history. Considering the many variables analysed in the different nations, trans-national differences are often present for a given specific issue. However, with regard to the core issues, no significantly different patterns appear to emerge for any given nation. The current attitude towards the disorder displays an increased insight about the condition on the part of patients. There is also a definite trend towards an improvement in their perception of the disease with an evident minor or reduced impact of the disorder on the life-style of patients. CONCLUSIONS: Globally, the data indicate that in all participating countries there has been, in recent years, a consistent improvement in the 'perceived quality of life' of the 'informed patient' with an increased insight into the condition and an enhanced rate of compliance. Nevertheless, a lot still needs to be done to markedly improve the 'social functioning' and the 'social integration' of those who suffer from BD. The data reported underline some of the issues that still represent a truly onerous burden for BD patients in whichever European country they may live. PMID- 15541065 TI - The current status of psychological treatments in bipolar disorders: a systematic review of relapse prevention. AB - OBJECTIVE: This paper reviews published randomized-controlled treatment trials of psychological therapies added to standard psychiatric treatment versus medication and standard psychiatric treatment alone to explore whether adjunctive psychotherapy reduces relapse rates in individuals with bipolar disorders (BDs). METHOD: Relapse rates were calculated for individual trials that met inclusion criteria and then pooled odds ratios were calculated using meta-analytic techniques. RESULTS: The majority of studies quoted demonstrate that individuals receiving psychological treatments had significantly fewer relapses. The length of therapy required was between 10 and 20 h over 6-9 months and the models of effective therapies had many shared characteristics in terms of style and content. CONCLUSIONS: Adjunctive psychological treatments for individuals with BDs are acceptable and feasible and reduce relapse risk. There are relatively few differences in the benefits that accrue from the different therapy models. It is now important to explore whether they have added value in terms of additional health gains and social functioning compared with standard treatment approaches. PMID- 15541066 TI - A collaborative approach to the treatment alliance in bipolar disorder. AB - OBJECTIVES: The treatment alliance is the arena in which psychopharmacological and other therapeutic interventions occur. The nature and quality of the treatment alliance may affect adherence to treatment and the realization of the benefits of effective pharmacological treatment in clinical practice. It is an area that has attracted little systematic study, despite the available evidence suggesting that it plays a measurable role in clinical outcomes. METHODS: A literature search was undertaken using Medline, Ovid, Psychinfo and Science Direct from 1975 to 2004. The following key words were used: bipolar disorder, patient adherence, non-adherence to medication, compliance, doctor-patient relationship, doctor-patient communication, treatment alliance, therapeutic alliance, chronic illness management, collaborative care, self-management, health beliefs, self-efficacy, self-determination, autonomy support, motivational interviewing. RESULTS: Psychosocial interventions have demonstrated positive effects on adherence problems. Studies of the impact of the treatment alliance on outcomes in mental illness highlight the possibilities of fruitful research in this area in bipolar disorder. Different theoretical models of changing health related behaviour may inform approaches to the treatment alliance. CONCLUSIONS: Results suggest the usefulness of a collaborative approach to the treatment alliance. Attention needs to be given to developing intervention models that target modifiable risk factors for non-adherence and address patient, clinician and illness related variables to enhance medication adherence in the treatment alliance. Refinement of these models through controlled evaluation in real world settings may lead to integration in health care delivery systems. PMID- 15541067 TI - Guidelines for maintenance treatment of bipolar disorder: are there discrepancies between European and North American recommendations? AB - Practice policies and guidelines for the long-term management of bipolar patients have appeared in many parts of Europe and North America. Although recommendations in most areas do concur remarkable differences are apparent both regarding diagnostic practice and pharmacological management. Differences among recommendations point towards professional and cultural differences between Europe and North America but also towards areas with unresolved research questions and lack of scientific evidence. PMID- 15541068 TI - Rapid cycling bipolar disorder: historical overview and focus on emerging treatments. AB - OBJECTIVE: Rapid cycling bipolar disorder presents a significant challenge with respect to treatment. The cyclical nature of bipolar disorder has been well recognised for over a century, and following Dunner and Fieve's landmark paper in 1974, investigators have increasingly turned their attention to issues such as the definition of rapid cycling, demographic characterisation, treatment response and pharmacologic intervention. METHODS: A literature search using Medline was performed, and selected articles which consider important developments in the definition, demographics and course of rapid cycling are reviewed. In addition, a systematic review of the literature published during the past 5 years (1999-2004) relating to treatment was conducted. RESULTS: Relevant articles are reviewed. CONCLUSIONS: This review highlights the important developments in our understanding of rapid cycling bipolar disorder, and focuses particularly on the recent literature regarding treatment. PMID- 15541069 TI - Bipolar depression: phenomenological overview and clinical characteristics. AB - OBJECTIVES: There has been increasing interest in the depressed phase of bipolar disorder (bipolar depression). This paper aims to review the clinical characteristics of bipolar depression, focusing upon its prevalence and phenomenology, related neuropsychological dysfunction, suicidal behaviour, disability and treatment responsiveness. METHODS: Studies on the prevalence of depression in bipolar disorder, the comparative phenomenology of bipolar and unipolar depression, as well as neuropsychology and brain imaging studies, are reviewed. To identify relevant papers, a literature search using MEDLINE and PubMed was undertaken. RESULTS: Depression is the predominant mood disturbance in bipolar disorder, and most frequently presents as subsyndromal, minor or dysthymic depression. Compared with major depressive disorder (unipolar depression), bipolar depression is more likely to manifest with psychosis, melancholic symptoms, psychomotor retardation (in bipolar I disorder) and 'atypical' symptoms. The few neuropsychological studies undertaken indicate greater impairment in bipolar depression. Suicide rates are high in bipolar disorder, with suicidal ideation, suicide attempts and completed suicides all occurring predominantly in the depressed phase of this condition. Furthermore, the depressed phase (even subsyndromal) appears to be the major contributant to the disability related to this condition. CONCLUSIONS: The significance of the depressed phase of bipolar disorder has been markedly underestimated. Bipolar depression accounts for most of the morbidity and mortality due to this illness. Current treatments have significant limitations. PMID- 15541070 TI - Changes in neuronal activation in patients with bipolar disorder during performance of a working memory task. AB - OBJECTIVES: Several lines of evidence suggest that deficits in cognition persist in bipolar patients during periods of euthymia. Working memory impairment has been observed in euthymic bipolar patients and noted to be a significant source of functional deficits in psychiatric disorders. Functional changes associated with these cognitive deficits however, remain poorly understood. We hypothesized that patients with bipolar disorder would demonstrate changes in neuronal activation in specific regions forming part of the working memory network. METHODS: Fifteen euthymic bipolar patients and fifteen age- and gender-matched healthy controls were recruited. Subjects participated in fMRI scans during which a two-back working memory task alternated with a zero-back control/attention task using a block-design paradigm. Groups were analyzed separately, and intergroup comparisons were made using an exploratory, voxel-by-voxel analysis. RESULTS: Bipolar patients performed more poorly on the cognitive tasks than did healthy controls (F = 3.77, p = 0.04). After covarying for task performance and reaction time, bipolar patients demonstrated significantly greater activation than healthy subjects in several regions including the fronto-polar prefrontal cortex, temporal cortex, basal ganglia, thalamus, and posterior parietal cortex. No areas showed a significant decrease in activation, compared with healthy controls. CONCLUSIONS: Our findings suggest that decreased working memory performance in bipolar patients reflects specific neurofunctional deficits. These deficits may represent primary areas of neuropathology or be secondary to neuropathology elsewhere in the working memory network. Continued research utilizing other imaging modalities may further clarify the underlying neuropathology involved in these cognitive deficits. PMID- 15541071 TI - A functional MRI study of working memory task in euthymic bipolar disorder: evidence for task-specific dysfunction. AB - OBJECTIVES: Even when euthymic bipolar disorder patients can have persistent deficits in working memory, but the neural basis of this deficit remains unclear. We undertook an functional magnetic resonance imaging investigation of euthymic bipolar disorder patients performing two working memory paradigms; the two-back and Sternberg tasks, selected to examine the central executive and the phonological loop respectively. We hypothesized that neuronal dysfunction would be specific to the network underlying the executive rather than the phonological loop component of working memory. METHODS: Twelve right-handed euthymic bipolar I males receiving lithium carbonate monotherapy were matched with 12 controls. The two-back task comprised a single working memory load contrasted with baseline vigilance condition. The Sternberg paradigm used a parametric design incorporating variable working memory load with fixed delay between presentation of an array of items to be remembered and a target item. Functional activation data were acquired during performance of the tasks and were analysed to produce brain activation maps representing significant group differences in activation (ANOVA). Load-response curves were derived from the Sternberg task data set. RESULTS: There were no significant between-group differences (t-test) in performance of the two-back task, or in 2 x 5 group by memory load ANOVA for the performance data from Sternberg task. In the two-back task, compared with controls bipolar disorder patients showed reductions in bilateral frontal, temporal and parietal activation, and increased activations with the left precentral, right medial frontal and left supramarginal gyri. No between-group differences were observed in the Sternberg task at any working memory load. CONCLUSIONS: Our findings support the notion that, in euthymic bipolar disorder, failure to engage fronto-executive function underpins the core neuropsychological deficits. PMID- 15541072 TI - Diagnosis in dysmorphology: clues from the skin. AB - Dysmorphology is the study of abnormal patterns of human development. A recurrent and recognizable combination of physical and behavioural abnormalities makes up a syndrome. Accurate recognition and diagnosis of syndromes is important because it influences medical management of patients, provides information about prognosis, and allows for genetic counselling including accurate estimation of genetic risk within families and, where possible, prenatal diagnosis. This review examines the diagnostic process in dysmorphology and indicates how skin signs may provide important clues to the clinician. PMID- 15541073 TI - Increase of laminin 5 synthesis in human keratinocytes by acute wound fluid, inflammatory cytokines and growth factors, and lysophospholipids. AB - BACKGROUND: Laminin 5 is known to induce the adhesion, spreading and migration of human keratinocytes. In skin wound healing, laminin 5 deposition beneath migrating keratinocytes occurs early and is followed by the formation of hemidesmosomes and then basement membrane. OBJECTIVES: To identify factors that regulate the synthesis and secretion of laminin 5 by human keratinocytes during acute wound healing. METHODS: Laminin 5 synthesis by human keratinocytes was determined by a specific sandwich enzyme-linked immunosorbent assay. To determine the total amount of laminin 5 synthesized, laminin 5 deposited on culture dishes and inside cells was solubilized by detergent solution and determined separately from conditioned medium, and the total laminin 5 synthesis was calculated. A quantitative polymerase chain reaction method was used to measure the expression levels of laminin 5 genes, LAMA3, LAMB3 and LAMC2, which correspond to the alpha3, beta3 and gamma2 chains of laminin 5. We also examined the effects of lysophospholipids, proinflammatory cytokines and growth factors, which are components in acute wound fluids, on laminin 5 synthesis in keratinocytes. RESULTS: Human acute wound fluid at days 1, 2 and 3 stimulated laminin 5 synthesis in cultured human keratinocytes in a concentration-dependent manner, although findings are restricted to one case. Human serum also increased laminin 5 production by human keratinocytes as strongly as the wound fluid did, suggesting that the major active components in acute wound fluid may be derived from those in human serum. Lysophospholipids such as lysophosphatidic acid (LPA), lysophosphatidylcholines (LPCs) and sphingosine-1-phosphate (S1P) increased laminin 5 synthesis in a concentration-dependent manner. Among growth factors, epidermal growth factor, insulin-like growth factor-1, interferon-gamma and keratinocyte growth factor increased laminin 5 production in keratinocytes, while platelet-derived growth factor, hepatocyte growth factor and basic fibroblast growth factor were ineffective. Although interleukin-1alpha had no effect, transforming growth factor (TGF)-alpha, tumour necrosis factor (TNF)-alpha and TGF-beta1 also stimulated laminin 5 synthesis, and TGF-alpha and TGF-beta1 showed a synergistic effect. Neutralizing antibodies to TGF-alpha and TGF-beta1 markedly inhibited the enhanced laminin 5 synthesis by human serum, suggesting that TGF alpha and TGF-beta1 are important components to increase laminin 5 in human serum. In line with the increase of laminin 5 synthesis, the expression levels of all three laminin 5 genes were also augmented by TGF-alpha and TGF-beta1. CONCLUSIONS: Laminin 5 synthesis in human keratinocytes was augmented by inflammatory cytokines and growth factors such as TGF-alpha, TGF-beta1 and TNF alpha, and lysophospholipids such as S1P, LPA and LPCs, which are supposed to be present in acute wound fluid. The increased laminin 5 protein in the wound area presumably enhances wound repair by stimulating adhesion and migration of keratinocytes on the wound bed and by facilitating basement membrane formation at the dermal-epidermal junction. PMID- 15541074 TI - The quality of human skin xenografts on SCID mice: a noninvasive bioengineering approach. AB - BACKGROUND: Animal models are important tools for studies in skin physiology and pathophysiology. Due to substantial differences in skin characteristics such as thickness and number of adnexa, the results of animal studies cannot always be directly transferred to the human situation. Therefore, transplantation of human skin on to SCID (severe combined immunodeficiency) mice might offer a promising tool to perform studies in viable human skin without the direct need for human volunteers. OBJECTIVES: To characterize the physiological and anatomical changes of a human skin transplant on a SCID animal host. METHODS: In this study human skin was transplanted on to 32 SCID mice and followed for 6 months. Barrier function was assessed by transepidermal water loss (TEWL; tewametry) and moisture content of the stratum corneum was studied by measurement of electrical capacitance (corneometry). RESULTS: The results showed considerable deviations of TEWL values and skin hydration between the grafts and human skin in vivo. The human skin showed epidermal hyperkeratosis and moderate sclerosis of the corium 4 and 6 months after transplantation on to SCID mice. CONCLUSIONS: Our results indicate that human skin does not completely preserve its physiological and morphological properties after transplantation on to SCID mice. Therefore, results from experiments using this model system need to be discussed cautiously. PMID- 15541075 TI - Topically applied S-nitrosothiol-containing hydrogels as experimental and pharmacological nitric oxide donors in human skin. AB - BACKGROUND: Nitric oxide (NO) has a wide range of functions in the skin, and topical NO donors have several potential clinical applications. However, currently available donors are either unstable on the skin surface, release low concentrations of NO, or have a short duration of action. Endogenous S nitrosothiols (RSNOs) store and transport NO within the body and can be used as exogenous sources of NO. OBJECTIVES: To study in vitro and in vivo the chemical and biological behaviour of two RSNO species, S-nitrosoglutathione (GSNO) and S nitroso-N-acetylcysteine (SNAC), in an easily applied hydrogel, and to correlate dermal nitrite concentration with erythema following application of the RSNOs. To assess the suitability of GSNO and SNAC as biologically effective NO donors for clinical research and as potential therapeutic agents. METHODS/PATIENTS: GSNO (0.3 mol g(-1)) and SNAC (0.6 mol g(-1)) were incorporated in Synperonic F-127 hydrogels (Uniquema, Belgium). The in vitro kinetics of decomposition were measured by spectrophotometry at 37 degrees C. The RSNO-containing hydrogels were applied to the forearm skin of eight subjects. Blood flow was measured by laser Doppler for 3 h following application of NO donors and dermal nitrite simultaneously measured in microdialysate in four subjects. RESULTS: The mean peak blood flow achieved was 250. At blood flow values of < 250, dermal nitrite correlated closely with blood flow and could be defined by the equation: blood flow = (nitrite concentration x 0.66) + 120, (P = 0.013). At higher blood flows there was a paradoxical fall in dermal nitrite concentration. CONCLUSIONS: Topical RSNOs produce a consistent, sustained and biologically effective release of NO on human skin in vivo, which offers advantages over currently available topical NO donors. Dermal nitrite concentration--the oxidation product of NO--is directly correlated with blood flow at low and moderate levels of blood flow. At high levels of blood flow, there is a reduction in dermal nitrite, which is presumed to be due to increased blood scavenging. PMID- 15541076 TI - Ovariectomy is sufficient to accelerate spontaneous skin ageing and to stimulate ultraviolet irradiation-induced photoageing of murine skin. AB - BACKGROUND: Wrinkling and sagging of the skin during photoageing is physiologically associated with diminished elasticity, which can be attributed to increased fibroblast-derived elastase activity. This degrades the dermal elastic fibres needed to maintain the three-dimensional structure of the skin. We previously reported that ovariectomy accelerates ultraviolet (UV)B-induced wrinkle formation in rat hind limb skin by altering the three-dimensional structure of elastic fibres. OBJECTIVES: In this study, we used hairless mice to assess the effects of ovariectomy with or without chronic UVA or UVB radiation on sagging and wrinkling of skin, on the elasticity of skin, as well as on matrix metalloproteinase activities in the skin. METHODS: Ovariectomies or sham operations were performed on 6-week-old female ICR/HR hairless mice. RESULTS: Even in the ovariectomy group without UV irradiation, the skin elasticity was significantly decreased during the 3-13 weeks after ovariectomy, which was accompanied by a significant increase in elastase activity in the skin. After UVA or UVB irradiation, skin elasticity was significantly decreased to a greater extent in the ovariectomy group than in the sham operation group, and this was accompanied by a reciprocal increase in elastase activity but not in the activities of collagenases I or IV in the skin. Consistent with the decreased skin elasticity, UVA irradiation for 12 weeks elicited more marked sagging in the ovariectomy group than in the sham operation group. UVB irradiation for 12 weeks also induced more marked wrinkle formation in the ovariectomy group than in the sham operation group. CONCLUSIONS: These results suggest that ovariectomy alone is sufficient to accelerate skin ageing and to increase UV sensitivity, which results in the further deterioration of the skin and photoageing, and may account for the accelerated skin ageing seen in postmenopausal women. PMID- 15541077 TI - Keratinocytes in the depigmented epidermis of vitiligo are more vulnerable to trauma (suction) than keratinocytes in the normally pigmented epidermis, resulting in their apoptosis. AB - BACKGROUND: Vitiligo may develop following minor physical trauma. However, in autologous epidermal grafting, depigmentation of the donor (normally pigmented) site from a suction blister is rare, even in cases displaying failure of repigmentation at the recipient (depigmented) site. OBJECTIVES: To examine whether the suction procedure is more likely to damage keratinocytes in the depigmented than in the normally pigmented epidermis of vitiligo, and to determine what kind of damage occurs to the keratinocytes. METHODS: Paired roofs of suction blisters from five patients with generalized vitiligo, five with localized and seven with segmental type, were used for the study. Multiple new lesions developed in two of the five patients with the generalized type. Apoptosis of keratinocytes in the epidermis was determined by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end labelling (TUNEL) staining, with immunohistochemistry for Bax and active caspase 3. Expression of Bcl-2, Bax, FLIP and p53, activation of caspases 3, 8 and 9, and cleavage of poly(adenosine diphosphate ribose) polymerase (PARP) in the epidermis were analysed by Western blotting in four patients with each type. RESULTS: Apoptotic keratinocytes, which stained with TUNEL and anti-Bax and antiactive caspase 3 antibodies, were scattered in the blistered epidermis, mainly in the lower portions. The depigmented epidermis displayed significantly more apoptotic keratinocytes than the normally pigmented epidermis. The numerical difference between the paired epidermides was related to the disease activity and not to the type of lesions. The number of apoptotic keratinocytes in the normally pigmented epidermis was as high as that in the depigmented epidermis in the two patients with active generalized type vitiligo. Expression of Bax and p53 in the depigmented epidermis was higher than in the normally pigmented epidermis, whereas expression of FLIP was lower. In addition, the activation of caspases 3, 8 and 9, and cleavage of PARP, were increased in the depigmented compared with the normally pigmented epidermis. The degree of difference in expression and activation was parallel to the results of the TUNEL assay. CONCLUSIONS: The keratinocytes in the depigmented compared with the normally pigmented epidermis of vitiligo may become apoptotic more easily after suction. PMID- 15541078 TI - Development of a novel ELISA system for detection of anti-BP180 IgG and characterization of autoantibody profile in bullous pemphigoid patients. AB - BACKGROUND: The NC16A immunodominant region of the bullous pemphigoid (BP) antigen BP180 has been used to develop several enzyme-linked immunosorbent assays (ELISAs) as diagnostic tools for BP autoantibody detection. OBJECTIVES: Because BP180 autoantibody reactivity is not restricted to NC16A, we have investigated the possibility of developing an ELISA based on selected epitopes additional to this immunodominant region. METHODS: Initially 78 BP sera were tested using an NC16A ELISA and IgG reactivity was detected in 64 BP sera (82%). The 14 NC16A negative BP sera were then analysed by immunological screening against seven BP180-specific epitopes. Recombinant phages displaying BP180 epitopes were grown as plaques, blotted onto a nitrocellulose filter and incubated with BP sera. RESULTS: Three and five NC16A-negative BP sera reacted with epitopes AA 1080-1107 and AA 1331-1404 of the BP180 ectodomain, respectively. Thus, a novel ELISA with GST-1080 and GST-1331 (GST-1080/1331) was developed: 32 of 78 BP sera (41%) proved positive by this assay. The combined use of ELISAs with GST-NC16A and GST 1080/1331 detected IgG reactivity in 72 of 78 BP sera, increasing the sensitivity from 82% to 92%. In addition, autoreactivity against the three extracellular epitopes appeared to be related to the presence of both skin and mucosal involvement as assessed by Fisher's exact probability test. CONCLUSIONS: Our findings further characterize the autoimmune response in BP by identifying a subgroup of NC16A-negative patients who react with different BP180 extracellular epitopes. The developed ELISA system appears more sensitive than the ELISA based on NC16A alone and also informative about the epitope profile of BP patients. PMID- 15541079 TI - Pregnancy and obstetrical outcomes in pseudoxanthoma elasticum. AB - BACKGROUND: Pseudoxanthoma elasticum (PXE) is a genetic multisystem disorder characterized by calcified dystrophic elastic fibres in skin, retina and arteries. Much of the earlier literature on pregnancy in PXE contained reports of severe complications, leading some healthcare providers to advise women with PXE against becoming pregnant and some women with PXE to avoid pregnancy. OBJECTIVES: To evaluate the obstetrical outcomes and the incidence of pregnancy complications in women with PXE and to determine if pregnancy is associated with an adverse effect on the course of the disease. METHODS: Women with PXE (n = 407) answered detailed questionnaires regarding reproductive history and pregnancy as well as the course of their disease. The frequency of reported pregnancy outcomes and complications was determined. Severity indices for the major clinical manifestations of PXE were developed and correlated with gravidity of affected women aged 40 years or over. RESULTS: Among the 306 respondents with PXE who had ever been pregnant, there were 795 pregnancies. Of these, 83% ended in live births and 1% in stillbirth. The median birth weight was within the normal range and the incidence of low birth weight for gestation was low. Hypertension occurred in 10% of pregnancies, gastric bleeding and retinal complications in < 1%, and 12% of pregnancies were associated with worsening of skin manifestations. There was no effect of gravidity and clinical severity on cutaneous (P = 0.07), ocular (P = 0.59) or cardiac (P = 0.42) manifestations of PXE in women aged 40+ years, nor did ever having been pregnant adversely affect these clinical severity indices. Of the 101 women who had never been pregnant, 17% made the decision because they were advised against becoming pregnant by a healthcare professional and 11% did not become pregnant because they feared an adverse outcome either in their pregnancy or disease. CONCLUSIONS: PXE is not associated with markedly increased fetal loss or adverse reproductive outcomes. The incidence of gastric bleeding, although probably higher than in the unaffected population, is much lower than previously reported, and retinal complications are uncommon. Although a few pregnancies were associated with worsening of skin manifestations, there was no correlation of either gravidity or ever having been pregnant with ultimate severity of skin, ocular or cardiovascular manifestations. There is no basis for advising women with PXE to avoid becoming pregnant, and most pregnancies in PXE are uncomplicated. PMID- 15541080 TI - Rapid healing of intractable diabetic foot ulcers with exposed bones following a novel therapy of exposing bone marrow cells and then grafting epidermal sheets. AB - BACKGROUND: Diabetic foot ulcers with exposed bones commonly result in amputation. OBJECTIVES: To determine whether exposure of bone marrow cells and subsequent grafting of epidermal sheets accelerates healing and reduces the need for amputation. METHODS: Thirty-eight patients with chronic wounds caused by diabetes mellitus were enrolled in this study. Epidermal sheets obtained from suction blisters of each patient were grafted on to diabetic foot ulcers without exposed bones (n = 10) and were compared with the standard treatment of local wound care, debridement with a scalpel when indicated, bed rest and parenteral antibiotics (n = 8). In another group of patients, diabetic wounds with exposed bones were treated either with the standard procedure (n = 9) or with a newly developed experimental procedure (n = 11). In that new procedure, the affected bone was initially exposed by debridement with a scalpel, followed by partial excision with a bone scraper until fresh bleeding was observed from the exposed bone. The lesions were then immediately covered with an occlusive dressing, and finally the wound was covered with an epidermal graft of skin harvested from suction blisters. Patients in each group were matched with their counterparts by age, sex, wound size, wound infection and wound duration, to compare the time needed for total skin repair and rates of amputation. RESULTS: Epidermal grafting significantly accelerated the healing of diabetic foot ulcers (P = 0.042) without exposed bones, with site-specific differentiation. The newly developed combination therapy resulted in the healing of all diabetic ulcers with exposed bones without the occurrence of osteomyelitis or the necessity for amputation (P < 0.0001). CONCLUSIONS: Our study indicates that early aggressive debridement of diabetic foot ulcers with exposed bones down to a bleeding vascularized base and then grafting epidermal sheets significantly improves healing and reduces the rate of amputation. PMID- 15541081 TI - Digital image analysis for diagnosis of cutaneous melanoma. Development of a highly effective computer algorithm based on analysis of 837 melanocytic lesions. AB - BACKGROUND: Digital image analysis has been introduced into the diagnosis of skin lesions based on dermoscopic pictures. OBJECTIVES: To develop a computer algorithm for the diagnosis of melanocytic lesions and to compare its diagnostic accuracy with the results of established dermoscopic classification rules. METHODS: In the Department of Dermatology, University of Tuebingen, Germany, 837 melanocytic skin lesions were prospectively imaged by a dermoscopy video system in consecutive patients. Of these lesions, 269 were excised and examined by histopathology: 84 were classified as cutaneous melanomas and 185 as benign melanocytic naevi. The remaining 568 lesions were diagnosed by dermoscopy as benign. Digital image analysis was performed in all 837 benign and malignant melanocytic lesions using 64 different analytical parameters. RESULTS: For lesions imaged completely (diameter < or = 12 mm), three analytical parameters were found to distinguish clearly between benign and malignant lesions, while in incompletely imaged lesions six parameters enabled differentiation. Based on the respective parameters and logistic regression analysis, a diagnostic computer algorithm for melanocytic lesions was developed. Its diagnostic accuracy was 82% for completely imaged and 84% for partially imaged lesions. All 837 melanocytic lesions were classified by established dermoscopic algorithms and the diagnostic accuracy was found to be in the same range (ABCD rule 78%, Menzies' score 83%, seven-point checklist 88%, and seven features for melanoma 81%). CONCLUSIONS: A diagnostic algorithm for digital image analysis of melanocytic lesions can achieve the same range of diagnostic accuracy as the application of dermoscopic classification rules by experts. The present diagnostic algorithm, however, still requires a medical expert who is qualified to recognize cutaneous lesions as being of melanocytic origin. PMID- 15541082 TI - Fruit acids and sodium hydroxide in the food industry and their combined effect with sodium lauryl sulphate: controlled in vivo tandem irritation study. AB - BACKGROUND: Cutaneous exposure to a variety of irritants has been extensively studied in recent years. Nevertheless, knowledge of the induction of irritant dermatitis, especially by mild irritants at low doses and for a short duration of exposure, is still incomplete. OBJECTIVES: To quantify the irritant effects and barrier disruption properties of ascorbic acid (ASC), acetic acid (ACA) and sodium hydroxide (NaOH), particularly in combination with an anionic detergent, sodium lauryl sulphate (SLS). METHODS: In a tandem repeated irritation test, the irritants were applied for 30 min twice daily for 4 days to the skin of the mid back of 19 healthy volunteers of both sexes. We used bioengineering techniques for measurement of transepidermal water loss (TEWL) and skin colour reflectance, as well as visual scoring. RESULTS: Repeated application of ASC and ACA caused a moderate increase in TEWL and erythema. The sequential application of ASC or ACA and SLS enhanced these effects. NaOH induced a strong reaction when applied both occlusively and nonocclusively as well as in combination with SLS, with an early onset of the inflammatory signs, leading to discontinuation of the application on the third day in most of the test fields. Notably, the irritant effect of NaOH was not as marked when applied sequentially with SLS. CONCLUSIONS: Our results demonstrate that concurrent application of an anionic detergent and a mild acidic irritant can lead to disruption of the barrier function which, although not additive, is still considerable. The combined application of SLS and mild acids does not prevent SLS-induced irritation. Furthermore, we showed that NaOH in low concentrations may also act as a potent irritant but that its effect is not enhanced by SLS. The necessity of adequate skin protection and reduction of contact with substances that are potentially barrier disruptive and irritant, e.g. in the food industry, is emphasized, not only when handling detergents, but also when processing food products. PMID- 15541083 TI - Depigmented extramammary Paget's disease. AB - BACKGROUND: Depigmented extramammary Paget's disease (EMPD) has been reported in a few cases. Depigmented macules or patches may be the only presenting sign or may coexist with the classical erythematous lesions. OBJECTIVES: To investigate the occurrence rate and clinical presentation of depigmentation in EMPD. METHODS: All pathology-proven cases of EMPD diagnosed in our department during 1990-2003 were retrieved. The clinical photographs were reviewed for evidence of local depigmentation. The pathological diagnosis of EMPD in the whitish lesions was confirmed by positive expression of cytokeratin 7 or carcinoembryonic antigen, and/or the presence of intracytoplasmic mucin. RESULTS: Of 19 cases of EMPD, six (30%) manifested depigmented lesions which were confirmed to be EMPD pathologically. In two patients, the hypopigmentation was associated with erythematous lesions at the initial presentation. In four others, the depigmentation developed later as local recurrence after excision, cryotherapy, photodynamic therapy or radiotherapy. The progressive enlargement of the depigmentation and the appearance of separate new white lesions in these four cases suggested that the localized depigmentation was unlikely to be simple postinflammatory hypopigmentation. CONCLUSIONS: Our study suggests that depigmented EMPD may not be rare. Localized depigmentation in the genital area can be an early sign of EMPD and its local recurrence. In patients with an established diagnosis of EMPD, appearance of new white lesions and continuous enlargement of depigmented patches should not be dismissed as simple treatment induced postinflammatory hypopigmentation or another type of hypopigmented lesion without biopsy confirmation. PMID- 15541084 TI - Comparative genomic hybridization in epithelioid sarcoma. AB - BACKGROUND: Epithelioid sarcoma is a rare mesenchymal neoplasm of unknown histogenesis. Data on genome-wide surveys for chromosomal aberrations in epithelioid sarcoma are limited. OBJECTIVES: To investigate genetic aberrations in epithelioid sarcoma. METHODS: We analysed seven cases of epithelioid sarcoma (classic type, three cases and proximal type, four cases) by comparative genomic hybridization (CGH), and correlated findings with the results of additional immunohistochemical study. RESULTS AND CONCLUSIONS: CGH analysis showed DNA copy number changes at one to five different genomic sites in six of seven cases (86%). The majority of the changes were gains. The most frequent gain was at 22q (six cases). Other recurrent changes include gains of 12q24-qter (four cases), 17 (four cases), and 5q32-qter (three cases). High-level homology was seen in chromosomal aberration in both types. In addition, expression of interleukin-2 receptorbeta, located in 22q, was revealed by immunohistochemical method in six cases with gain of 22q, suggesting it may play a role in epithelioid sarcoma tumorigenesis. PMID- 15541085 TI - Detection of epidermodysplasia verruciformis-associated human papillomavirus DNA in nongenital seborrhoeic keratosis. AB - BACKGROUND: DNA of epidermodysplasia verruciformis (EV)-associated human papillomaviruses (HPVs) has been widely detected in lesions of malignant skin tumours, benign tumours and other proliferative diseases of epithelial origin. OBJECTIVES: To investigate the presence of EV-associated HPV DNA in nongenital seborrhoeic keratosis (SK) and to elucidate the prevalence of distinct HPV genotypes. METHODS: We investigated HPV DNA in 55 nongenital SK biopsies, which were compared with 48 normal skin biopsies (healthy controls) using a nested polymerase chain reaction (PCR) using consensus primers CP65/CP70 and CP66/CP69. The positive PCR products were retracted and used to prepare recombination clones with T-vector. Distinct clones were analysed with endonucleases, and HPV genotypes were identified by direct sequencing. RESULTS: EV-associated HPV DNA was detected in 42 of 55 (76%) nongenital SK biopsies vs. only 13 of 48 (27%) healthy controls (chi2 = 22.087; P < 0.005). The prevalence was higher in patients with more than five lesions than in those with only one lesion (P < 0.05). Ten distinct HPV genotypes were detected in the nongenital SK biopsies: HPV 20, 23, 5, renal transplant recipient (RTR) X7, HPV 17, 37, 17b, RTRX4, RTRX4b and strain SK3. HPV 20 was found in 26 of 42 (62%) positive specimens, followed by HPV 23 (11 of 42, 26%) and HPV 5 (six of 42, 14%). Existence of multiple HPV genotypes was observed in 12 of 42 (29%) positive specimens. In healthy controls, five genotypes of EV-associated HPV (HPV 20, 23, 5, 17 and RTRX4) were detected, with the same predominant genotype of HPV 20 (five of 13, 38%). Several distinct HPV genotypes were found to coexist in four of 13 (31%) positive specimens. CONCLUSIONS: This study provides some evidence that EV associated HPVs might play a part in the pathogenesis of nongenital SK. PMID- 15541086 TI - Provocation testing in polymorphic light eruption using fluorescent ultraviolet (UV) A and UVB lamps. AB - BACKGROUND: Provocation testing is frequently performed during investigation of patients with suspected polymorphic light eruption (PLE). Techniques are not standardized between centres. OBJECTIVES: We sought to evaluate the efficacy of different fluorescent ultraviolet (UV) radiation lamps for provocation testing in PLE. METHODS: We analysed results in 68 patients referred consecutively for phototesting in whom a diagnosis of PLE seemed likely based on clinical history. Patients' case notes were reviewed and responses recorded to provocation testing on forearm skin over three consecutive days using broadband UVA, narrowband and broadband UVB lamps. RESULTS: A positive papular response to broadband UVA exposure was seen in 38 patients [56%, estimated 95% population confidence interval (CI) 43-67.9]. Thirty-four patients (50%) had a positive papular response to narrowband UVB exposure (95% CI 37.6-62.4). The probability of a positive provocation test following irradiation with both lamps was 80.9% (95% CI 69.5-89.4). From April 1999, 34 patients also had provocation testing with broadband UVB. Although six patients (18%) had a positive papular response, they all showed a positive response to one or both of the other lamp types. CONCLUSIONS: Provocation testing with fluorescent UVA and UVB lamps is a cheap and readily available method that can be used as a diagnostic aid to investigate patients with suspected PLE. Using both broadband UVA and narrowband UVB lamps for testing increases the likelihood of confirming the diagnosis than if either lamp is used alone. PMID- 15541087 TI - Pimecrolimus cream 1% vs. betamethasone 17-valerate 0.1% cream in the treatment of seborrhoeic dermatitis. A randomized open-label clinical trial. AB - BACKGROUND: Seborrhoeic dermatitis is a chronic inflammatory disease with remissions and exacerbations, characterized by erythema, scaling and pruritus primarily on the face, scalp and chest. Corticosteroids and antifungals are the mainstay of therapy. However, chronic use of corticosteroids is associated with side-effects such as skin atrophy and telangiectasia. Pimecrolimus, an inhibitor of calcineurin, has been used successfully in one patient with seborrhoeic dermatitis. OBJECTIVES: The objective of this randomized open-label clinical trial was to compare the efficacy and tolerability of pimecrolimus in comparison with a potent corticosteroid (betamethasone 17-valerate) in the treatment of seborrhoeic dermatitis. METHODS: Twenty patients with seborrhoeic dermatitis were included in this study, 11 patients in the pimecrolimus 1% cream group and nine patients in the betamethasone 17-valerate 0.1% cream group. Patients were instructed to use a thin layer of the study products twice daily at the lesional area and to discontinue treatment as soon as symptoms were absent. Clinical measures assessed were erythema, scaling and pruritus which were evaluated using a four-point scale (0-3). RESULTS: Both pimecrolimus and betamethasone were highly effective in the treatment of seborrhoeic dermatitis. Betamethasone reduced all three parameters, erythema, scaling and pruritus, faster than pimecrolimus, but the differences in reduction were not statistically significant. Relapses were observed more frequently and were more severe with betamethasone than with pimecrolimus. Moreover, pruritus was not observed after discontinuation of treatment from day 15 and beyond in the pimecrolimus group, whereas it was reported in most patients of the betamethasone group. This difference was statistically significant. CONCLUSIONS: It appears that pimecrolimus, a nonsteroidal topical treatment, may be an excellent alternative therapeutic modality for treating seborrhoeic dermatitis. PMID- 15541088 TI - Photodynamic therapy using meta-tetrahydroxyphenylchlorin (Foscan) for the treatment of vulval intraepithelial neoplasia. AB - BACKGROUND: Photodynamic therapy (PDT) has unique properties which make it suitable for the local treatment of superficial epithelial disorders; it has been suggested as a useful treatment for carcinoma in situ of the vulva. OBJECTIVES: To evaluate the effect of the systemic photosensitizing agent meta tetrahydroxyphenylchlorin (mTHPC or temoporfin; Foscan, Biolitec, Edinburgh, U.K.) in vulval intraepithelial neoplasia type III (VIN III). METHODS: PDT using mTHPC was performed in six patients with VIN III. A dose of 0.1 mg kg(-1) body weight mTHPC was injected intravenously and the area of VIN irradiated 96 h later with 652-nm light from a diode laser. Patients were reviewed 1 week, 6 months and 2 years following treatment. RESULTS: Patients experienced only minimal pain from the initial treatment but two patients subsequently developed severe pain at the treated site for up to 2 weeks following PDT. All patients developed oedema and slough formation at the treated site and one patient developed cellulitis. At 6 months two patients had developed small recurrences of VIN at the original site and one patient had an area of VIN at a new site. These were treated either with further PDT or with a small excision. At 2 years there was no recurrence of VIN at the original site in all patients reviewed. CONCLUSIONS: This small case series demonstrates that mTHPC-PDT is a useful initial treatment for VIN III. It is relatively selective, shows good cosmesis and conserves form and function. This is a major advantage over surgery. Repeat treatments are also possible, which is important in a condition such as VIN, which tends to be multifocal. Systemic mTHPC-PDT appears to have an advantage over topical 5-aminolaevulinic acid-PDT as the photosensitizer is distributed widely in areas of disease and consequently identifies foci which may not be apparent clinically but become evident when illuminated. PMID- 15541089 TI - Small diameter melanoma: a follow-up of the Norwegian Melanoma Project. AB - BACKGROUND: Melanoma prognosis is dependent upon early recognition and treatment. There is a need for good clinical guidelines that focus on the early signs of melanoma. The ABCD (asymmetry, border, colour and diameter) rule states that most melanomas are more than 6 mm in diameter. Critics crave a modification, arguing that small diameter melanomas are not infrequent. OBJECTIVES: The aim of the present study was to describe the frequency and prognosis of melanomas less than 7 mm in a clinical setting. METHODS: The Norwegian Melanoma Project was conducted as a multicentre, prospective study with inclusion criteria. Patients were recruited from five dermatological departments in Norway from 1990 to 1993. RESULTS: The frequency of small melanomas was 11.4% (18/158). One-third was in situ melanoma, the rest invasive with a median thickness of 0.8 mm. Four small melanomas were T2 lesions, with a Breslow thickness of more than 1 mm. One nodular T2 melanoma recurred locally 2 years after diagnosis and the patient died of distant metastasis only months later. CONCLUSIONS: The ABCD rule remains a practical guide for early recognition of melanoma. Clinicians must be aware of its limitations. PMID- 15541090 TI - Mild phenotype of familial cylindromatosis associated with an R758X nonsense mutation in the CYLD tumour suppressor gene. AB - Familial cylindromatosis is a rare dominantly inherited disease characterized by the development of multiple benign tumours of the skin appendages, including cylindromas, trichoepitheliomas and spiradenomas. The gene responsible was positionally cloned recently, and was designated CYLD. We describe a family with cylindromatosis, in which affected individuals have an inherited R758X nonsense mutation of CYLD. Affected members of this family manifest a relatively mild tumour phenotype; the largest tumour was only 30 mm in diameter. Thus far, there is no evident genotype-phenotype relationship in cylindromatosis, although the number of families reported with both phenotypic and genotypic data remains small. PMID- 15541091 TI - One-year follow-up of a lentigo maligna: first dermoscopic signs of growth. AB - We report a 64-year-old man with a pigmented lesion on his forehead, initially thought to be actinic lentigo. At follow-up 1 year later the lesion had increased in size and showed new areas of pigmentation. Dermoscopic observation and biopsy led to a diagnosis of lentigo maligna and the lesion was excised. The dermoscopic features indicative of early growth of lentigo maligna are identified and discussed. PMID- 15541092 TI - Leukocytapheresis treatment for pyoderma gangrenosum. AB - A 42-year-old man presented with painful erythema with pustules and multiple small ulcers on the shins. He had suffered from ulcerative colitis (UC) and received oral glucocorticosteroid and salicylazosulfapyridine therapies for 7 years. Biopsy of the lesion demonstrated mixed cellular infiltrates with dominant neutrophils. The patient was diagnosed with pyoderma gangrenosum (PG) and underwent leukocytapheresis (LCAP), an extracorporeal leucocyte removal therapy, once a week for 5 weeks without changing the doses of the oral medications. The skin lesions as well as clinical signs of UC rapidly improved after LCAP, and no recurrence was seen during a follow-up period. There were no major complications during LCAP. LCAP will provide an effective and safe tool for the treatment of PG. PMID- 15541093 TI - BOTOX delivery by iontophoresis. AB - We report two patients with severe palmar hyperhidrosis who responded to BOTOX delivered not by injection, the usual method of delivery, but by iontophoresis. The Botulinum molecule has been considered too large for delivery into the skin this way. However, other large peptides, both non-ionic and cationic, have been delivered successfully by this method, so we suspected that BOTOX could in fact be iontophoresed. Our saline-controlled treatment of these two patients with a small iontophoresis unit (Iomed Phoresor II) allowed small volumes of standard BOTOX dilutions to be used, and demonstrates that iontophoresis can indeed deliver BOTOX successfully. This has important therapeutic potential for the large number of patients with focal hyperhidrosis. They may be spared painful injections, and in more severe cases, invasive surgery. PMID- 15541094 TI - Successive development of cutaneous polyarteritis nodosa, leucocytoclastic vasculitis and Sweet's syndrome in a patient with cervical lymphadenitis caused by Mycobacterium fortuitum. AB - Mycobacterium fortuitum is a rapidly growing mycobacterium found in soil and water throughout the world. It can cause diseases in immunocompetent patients, usually resulting in localized skin and soft tissue infections. Cervical lymphadenitis caused by M. fortuitum is rare. We report a 46-year-old woman in whom skin lesions of cutaneous polyarteritis nodosa, leucocytoclastic vasculitis and Sweet's syndrome had successively developed before the diagnosis of cervical lymphadenitis caused by M. fortuitum was made. The skin lesions responded to colchicine and systemic corticosteroids but recurred intermittently. After establishment of the diagnosis, she received treatment with clarithromycin and ciprofloxacin. The cervical lymph nodes decreased in size 6 months later and no more new skin lesions were found. PMID- 15541095 TI - Mycobacterium chelonae infection successfully treated with oral clarithromycin and linezolid. PMID- 15541096 TI - Leukaemic macrocheilia in acute myeloblastic leukaemia. PMID- 15541098 TI - Unusual painful sclerotic plaques on the legs of a patient with late diagnosis of primary hyperoxaluria type I. PMID- 15541097 TI - Angiolymphoid hyperplasia with eosinophilia treated with anti-interleukin-5 antibody (mepolizumab). PMID- 15541099 TI - Chlorambucil as a steroid-sparing agent in bullous pemphigoid. PMID- 15541100 TI - Mycosis fungoides is not associated with hepatitis C virus infection. PMID- 15541101 TI - Angiolymphoid hyperplasia with eosinophilia successfully treated with imiquimod. PMID- 15541102 TI - Trichomegaly following treatment with gefitinib (ZD1839). PMID- 15541105 TI - A conversation between Darracott Vaughan and Michael Marberger. PMID- 15541106 TI - The era of serum prostate specific antigen as a marker for biopsy of the prostate and detecting prostate cancer is now over in the USA. PMID- 15541107 TI - Prostate cancer screening. PMID- 15541108 TI - Prostate-specific antigen testing for the early detection of prostate cancer. PMID- 15541109 TI - Future development of penile cancer services in the UK. PMID- 15541110 TI - Core competencies in urology: what are they and how are they best taught? PMID- 15541111 TI - Management of urethral stricture disease: developing options for surgical intervention. PMID- 15541112 TI - The role of the nonsurgical oncologist in the management of advanced transitional cell cancer. Part I: locally advanced disease. PMID- 15541113 TI - The role of the nonsurgical oncologist in the management of advanced transitional cell cancer. Part II: metastatic disease. PMID- 15541114 TI - Prostasomes: a role in prostatic disease? PMID- 15541115 TI - Evaluation of urinary extravasation and results after continence-preserving radical retropubic prostatectomy. AB - OBJECTIVE: To evaluate the feasibility of urinary catheter removal 10 days after a radical retropubic prostatectomy (RRP) by assessing the incidence of urinary extravasation and its effect on postoperative stricture and continence rates. PATIENTS AND METHODS: During a 4.5-year period, 619 patients undergoing RRP were evaluated. If no extravasation was detected on gravity cystography, the urinary catheter was removed 10 days after RRP. In patients with significant extravasation the catheter was left in place for 3 weeks. Overall stricture and continence rates were recorded in patients at 3, 6 and 12 months after surgery. RESULTS: There was extravasation during cystography in 29 patients (4.6%). At 3, 6 and 12 months, continence rates after catheter removal at 10 days were 74.9%, 87.9%, and 93.6%, respectively, while in the late-removal group they were 72.4%, 84.6% and 90.9%, respectively, with no significant difference between the groups. At 3 months the overall continence rate was 74.8% and at 12 months up to 93.5%. There was no difference in stricture rates between the groups, with an overall stricture rate of 0.7%. CONCLUSIONS: Catheter removal 10 days after RRP is feasible, giving excellent early and late continence rates, with low anastomotic stricture rates obtained using good surgical technique. Extravasation at 10 days was rare and with proper management did not influence the final results. PMID- 15541116 TI - A modified yeast assay used on archival samples of localized prostate cancer tissue improves the detection of p53 abnormalities and increases their predictive value. AB - OBJECTIVE: To determine the frequency and predictive value of p53 mutations in localized prostate cancer, comparing the accuracy of detection using immunohistochemistry (IHC) with a modified yeast assay, on archival tissue samples. MATERIALS AND METHODS: Prostate cancer tissue was obtained from 98 patients who had >/= 2 years of clinical follow-up after radical prostatectomy. DNA sequencing was used to verify the presence of p53 mutations in samples that were immunopositive or that gave evidence for p53 alterations using the yeast assay. The IHC and yeast findings were compared with patient outcome to determine the predictive value of these two test types. RESULTS: Fifty-five tumours (57%) were immunopositive, and 58 (59%) were positive using the yeast assay. Sequence confirmed p53 mutations occurred in 44 (45%) cases. The IHC protocol generated 49% (27/55) false-positive and 36% (15/42) false-negative results, and was 65% sensitive and 50% specific, with an overall accuracy of 57%. The yeast assay resulted in 24% (14/58) false-positive results with a specificity of 74% and an accuracy of 86%. When the p53 status of these patients was correlated with their clinical outcome, patients who had sequence-confirmed p53 mutations had a 2.6 fold greater failure rate (P = 0.026) and a 2.5-fold greater risk of dying from prostate cancer (P = 0.05). Notably, mutations in exon 6 predicted a six-fold increase in treatment failure (P = 0.043) and a 5.3-fold increase in the chance of dying from prostate cancer (P = 0.009). Abnormal yeast-assay findings gave similar predictive results to those obtained for DNA sequencing, while immunopositivity did not correspond to patient outcome. CONCLUSIONS: Mutations of p53 occurred in 45% of localized prostate cancers. These alterations have important prognostic implications. The yeast assay was more accurate for detecting p53 mutations than the IHC protocol used and, unlike IHC, the results of the yeast assay were predictive of patient outcome. PMID- 15541117 TI - Determinants of long-term quality of life and voiding function of patients treated with radical prostatectomy or permanent brachytherapy for prostate cancer. AB - OBJECTIVE: To assess the long-term quality of life (QoL) outcomes of three treatments for localized prostate cancer: radical prostatectomy (RP); brachytherapy monotherapy (BTM); and BT combined with external beam radiotherapy (BTC). PATIENTS AND METHODS: In August 2000, questionnaires were mailed to men with T1c-T3 adenocarcinoma of the prostate treated with either RP, BTM ((103)Pd monotherapy) or BTC. Questionnaires included validated outcome measures, i.e. the Functional Assessment of Cancer Therapy - General (FACT-G), American Urological Association Symptom Score (AUA-SS), Urinary Function Questionnaire for men after RP, and the Brief Sexual Function Inventory. Returned questionnaires were assessed using cross-sectional analysis. RESULTS: Data from 214 patients were included in the analysis (60 RP, 102 BTM and 52 BTC); the median follow-up was 18.8, 25.5 and 29.9 months, respectively. There were differences between both BT groups and the RP group in total AUA-SS and obstructive subscale symptom scores, with the former having worse symptom scores at a longer follow-up. Differences in overall QoL were not detected between groups using the total FACT-G but the BTC group generally had worse scores in the physical well-being subscale. The BT groups had higher continence rates with time after treatment. Sexual function was better with BT initially, but these differences did not persist at a longer follow-up. There were significant correlations between the FACT-G and the urinary symptom scores, and the degree of sexual function. CONCLUSIONS: Although patients treated with BTM and RP have a different spectrum of side-effects, their overall long-term QoL is similar, with urinary and sexual function being the primary determinants of this outcome. Men treated with BTC have a worse QoL. PMID- 15541118 TI - Threshold volumes for urological cancer surgery: a survey of UK urologists. AB - OBJECTIVE: To determine minimum threshold levels of activity set by surgeons for urological cancer surgery, and to relate threshold levels to stated current procedural volume. METHODS: In all, 307 consultant urological surgeons were sent a questionnaire asking them to state for four urological cancer operations of different complexity their current procedural volume; whether minimum volume thresholds per surgeon should be implemented; and if so, the level of such thresholds; 212 (69%) replied. RESULTS: For all four procedures >/= 75% of surgeons advocated the setting of a minimum volume threshold. Overall, surgeons set the highest thresholds for radical prostatectomy and the lowest for radical cystectomy with continent diversion. There was no significant association between either the principle of supporting minimum volume thresholds or the level of such a threshold and the number of years worked as a consultant surgeon. The level of surgeon-derived minimum thresholds increased with increasing surgeon procedural volume. CONCLUSION: Most surgeons supported the principle of setting minimum volume thresholds. These thresholds appear to be influenced by current procedural volume and by procedural complexity. By setting thresholds greater than their current volume, some surgeons implicitly indicate that their current volume is insufficient to maintain their surgical competency. PMID- 15541119 TI - Bleeding after transrectal ultrasonography-guided prostate biopsy: a study of 7 day morbidity after a six-, eight- and 12-core biopsy protocol. AB - OBJECTIVE: To compare, in a prospective study, bleeding (in three categories, i.e. haematuria, haematospermia and rectal) and consultations with the general practitioner (GP), after a six-, eight- or 12-core prostate biopsy, as data on whether taking more prostate core biopsies increases bleeding complications are not conclusive. PATIENTS AND METHODS: Over a 5-year period, patients undergoing outpatient transrectal ultrasonography (TRUS)-guided prostate biopsy (six, eight or 12-core biopsy) completed a self-administered questionnaire. The prevalence and duration of the three bleeding complications and GP or hospital visits for a biopsy-related complication were assessed and compared for the 7 days after biopsy. The contribution of local anaesthetic (LA) injection to bleeding rates was also assessed. RESULTS: Of 1384 patients biopsied, 1000 were given questionnaires and 884 (88%) forms were returned. Of these, 760 were suitable for analysis (307 after six-core, 325 eight-core and 128 12-core biopsies); 351 patients were given LA before biopsy. The prevalence of bleeding complications (six-, eight- and 12-core, respectively) was: haematuria 44%, 41% and 39%; haematospermia 13%, 16% and 12%; and rectal bleeding 17%, 26% and 27%. Rectal bleeding was significantly more prevalent in the eight- and 12-core groups (P = 0.0037 and 0.019). The duration of bleeding was not significantly greater in any biopsy group. Subgroup analysis showed no significant difference in the prevalence and duration of rectal bleeding after LA. About 5% of patients in each group consulted their GP because of a complication and 2.4% consulted because of bleeding. Three men with major complications required hospitalization, of which only one was caused by bleeding. CONCLUSIONS: Only rectal bleeding was more prevalent after taking more than six cores, but the duration was no greater. Giving LA did not affect the rectal bleeding rate. With all strategies the major complication and hospitalization rate was very low. PMID- 15541120 TI - Prostatic capsule- and seminal-sparing cystectomy for bladder carcinoma: initial results for selected patients. AB - OBJECTIVE: To evaluate the oncological outcome and functional results of prostate sparing cystectomy (PSC), proposed for treating bladder cancer, used since 1999 in our institution in an attempt to preserve male sexuality and to increase continence after cystectomy. PATIENTS AND METHODS: Between January 1999 and December 2001, 111 men were candidates for cystectomy; 42 were selected for a prostatic capsule- and seminal-sparing cystectomy with orthotopic urinary diversion. All patients had clinically organ-confined tumours (clinical stage /= 6 months duration with >/= 10 patients in each treatment arm. Data were extracted on study design, patient and treatment characteristics, urinary symptoms, urinary flow, adverse events and repeat treatment for BPH. RESULTS: Six studies were evaluated, involving 540 patients. The mean age (67.8 years), baseline symptom score (19.5), and peak urinary flow (PUF, 8.6 mL/s) did not differ by treatment group. The pooled mean urinary symptom score decreased by 65% with TUMT and 77% with TURP. The weighted mean (95% confidence interval) difference for the symptom score at the follow-up was 1.83 (-3.09 to -0.58) points, favouring TURP. The pooled mean PUF increased by 70% with TUMT and 119% with TURP. The weighted mean difference for the PUF at the follow-up was 5.37 (4.22-6.51) mL/s, favouring TURP. Retrograde ejaculation (57.6% vs 22.2%), transfusions (5.7% vs 0%) and re-treatment for strictures (relative hazard 9.76) were all significantly more common after TURP, but re treatment for BPH was significantly more common after TUMT (relative hazard 10.0). CONCLUSIONS: TUMT techniques are effective and safe short-term alternatives to TURP for treating BPH. However, TURP provided greater symptom and urinary flow improvements and fewer subsequent BPH treatments than TUMT. PMID- 15541123 TI - Long-term results of a self-expanding wallstent in the treatment of urethral stricture. AB - OBJECTIVE: To report the long-term outcome over 12 years of using the urethral Urolume wallstent (AMS, Minnetonka, MI, USA) for treating recurrent bulbar urethral stricture disease. PATIENTS AND METHODS: The case-notes of 60 consecutive men with urethral Urolume wallstents placed for treating recurrent bulbar strictures were reviewed retrospectively. Information was collected on patient demographics, stricture aetiology, stent-related complications and the need for further surgery to treat stent- or stricture-related complications. RESULTS: The mean (range) age of the men was 58 (32-76) years. The most common cause of stricture was iatrogenic, arising after previous endoscopic surgery or after an indwelling catheter (45%). Thirty-five men had complications, with re operation required in 27 (45%) of them. The most frequent nonsurgical complications were post-micturition dribble (32%) and recurrent urinary tract infections (27%). The most common surgical interventions required were transurethral resection of obstructing stent hyperplasia (32%), urethral dilatation or urethrotomy for stent obstruction or stricture (25%) and endoscopic litholapaxy for stent encrustation or stone (17%). CONCLUSIONS: The Urolume wallstent should only be used in patients who are unfit for or who refuse a bulbar urethroplasty. PMID- 15541124 TI - Treatment of stress urinary incontinence using a copolymer system: impact on quality of life. AB - OBJECTIVE: To investigate the effect on quality of life (QoL) of a novel system for treatment of stress urinary incontinence (SUI). PATIENTS AND METHODS: In an open, prospective, multicentre study, 42 women not previously treated by invasive therapy and with urodynamically verified SUI received 4 x 1.0 mL or 4 x 0.7 mL of non-animal stabilized hyaluronic acid/dextranomer copolymer injected transurethrally into the urethra via the Implacer device (Zuidex system, Q-Med AB, Uppsala, Sweden). QoL was assessed using the King's Health Questionnaire. The patients' perception of treatment benefit and the number of incontinence episodes/24 h were also investigated. RESULTS: There were significant improvements over baseline in seven of 10 domains of the King's Health Questionnaire at 3 months, and these improvements were sustained at 1 year. For the change from baseline to 1 year, eight of 10 domains showed a significant positive correlation with the number of incontinence episodes/24 h. In terms of treatment benefit, most of the women perceived an improvement at 1, 3, 6 and 12 months. Of the 18 women requiring re-treatment, most perceived an improvement at 3, 6 and 12 months. CONCLUSION: Treatment with the Zuidex system produced significant improvements over a year in both subjective QoL and objective incontinence measures, with a significant positive correlation between them. PMID- 15541125 TI - Urethral diverticula: a diagnostic dilemma. AB - OBJECTIVE: To review the urethral diverticula encountered in a tertiary-referral urogynaecology unit. PATIENTS AND METHODS: The case-notes of all women who had a diverticulectomy between March 1996 and May 2001 were reviewed. Demographic details, symptoms at presentation, duration of symptoms before diagnosis, investigations, operative details, postoperative complications and symptoms at follow-up were considered. RESULTS: In all, 18 women had had a urethral diverticulectomy. The median (range) time from presentation to diagnosis was 9.5 (2-96) months. The symptoms before surgery were variable; after surgery there were significant improvements in the symptoms of frequency, terminal dribbling and recurrent urinary tract infections (P < 0.05). There was no improvement in urgency, urge incontinence, nocturia and stress incontinence. Eleven of the 18 diverticula were palpable on vaginal examination. Video cysto-urethrography was used in 15 women and the diverticulum was seen in 14. In addition, seven women had additional lower urinary tract pathology. CONCLUSIONS: Urethral diverticula have no classical presentation; they often present with many symptoms and it is important that the diagnosis is not overlooked. Video cysto-urethrography is a good diagnostic test and allows the simultaneous evaluation of function of the whole of the lower urinary tract. PMID- 15541126 TI - A pilot randomized double-blind placebo-controlled trial on the use of antibiotics on urinary catheter removal to reduce the rate of urinary tract infection: the pitfalls of ciprofloxacin. AB - OBJECTIVE: To assess if a short course of antibiotics starting at the time of the removing a short-term urethral catheter decreases the incidence of subsequent urinary tract infection (UTI). PATIENTS AND METHODS: Patients across specialities with a urethral catheter in situ for >/= 48 h and /= 3 months and were assessed for any clinical and biochemical response. Of an initial group of 45 men (mean age 59.2 years) 38 completed the study. RESULTS: Most men presented with symptoms of sexual dysfunction, lack of energy and/or depression. There were differences before and after treatment only in bioavailable T (BT), with none in the levels of total T (TT). There was a strong correlation before and after treatment in the levels of luteinizing hormone and follicle-stimulating hormone, and a weak negative correlation between gonadotrophins and BT. Neither TT nor BT had predictive value for the treatment response. There was a trend to a correlation between BT levels and treatment success. Changes in serum prostate specific antigen were insignificant during the limited period. CONCLUSION: The lack of accurate methods for diagnosing SLOH suggests that a therapeutic trial of T supplementation is warranted in men in whom there are no contraindications. The 3-month period largely circumvents the placebo effect and has minimal risks for serious adverse effects (mostly in relation to prostate safety). This controversial position needs further evaluation with a larger cohort and other biochemical measurements. PMID- 15541130 TI - Anatomical studies of the neurovascular bundle and cavernosal nerves. AB - OBJECTIVE: To accurately define the anatomy of the neurovascular bundle (NVB) in men. MATERIALS AND METHODS: The NVB was microdissected in detail bilaterally in 12 fixed human male adult cadavers. The anatomy of the NVB and its relationship to surrounding pelvic structures was ascertained in each specimen. RESULTS: Previous reports of the anatomy of the NVB have not mentioned its levator ani and anterior rectal component. The anatomy of the cavernosal nerves is such that accurate graft anastomosis to proximal and distal cavernosal nerve segments is extremely difficult. CONCLUSION: The current anatomical description of the cavernosal nerve and NVB is inaccurate. PMID- 15541131 TI - A prospective study of the predictive power of spiral computed tomographic angiography for defining renal vascular anatomy before live-donor nephrectomy. AB - OBJECTIVE: To determine the accuracy of spiral computed tomography (CT) imaging of donor venous anatomy by comparing CT angiography (CTA) and operative findings, for both laparoscopic (LDN) and open donor nephrectomy. PATIENTS AND METHODS: LDN presents unique surgical challenges, particularly with complex venous or arterial anatomy. The limitations of surgical access, poor visibility of the superior and posterior borders of the renal vein during LDN, and the variability of venous anatomy in this region, contribute to the difficulty of LDN, underlining the importance of imaging beforehand. Forty live donors (mean age 46 years, sd 11; 65% female) were assessed by CTA before donation. Scans were reported by the same radiologist. The number and diameter of 'predicted' renal arteries, veins and renal vein tributaries were documented. The donor kidney was removed by two consultant surgeons, and after back-table perfusion the same details were recorded and taken as the 'reference' findings. Tributaries of <1 mm diameter were not recorded. The right kidney was retrieved in seven patients; 25 of the 40 kidneys were retrieved by LDN and the other 15 by open surgery. RESULTS: In all, 48 actual renal arteries were identified at nephrectomy; of these, 47 were predicted by CTA. Likewise, 41 actual renal veins were found at nephrectomy, 40 of which were predicted. The overall accuracy of spiral CTA in predicting the presence or absence of renal vein tributaries was 83% for gonadal and adrenal veins, and 75% for lumbar veins. There were seven false-negative lumbar veins found at nephrectomy; in these cases the CTA films were retrospectively examined, and five of these seven veins were identified. The predicted renal vein tributary diameter correlated poorly with the measured diameter at nephrectomy. CONCLUSIONS: Assessing potential renal donors before surgery with spiral CTA provides an accurate prediction of the presence or absence of the gonadal and adrenal vein, but is less accurate for predicting lumbar veins. This is especially pertinent as the posterior lumbar tributaries have the most intra individual variation, and are the most difficult to display and control at LDN. This highlights the need for meticulous dissection of the renal vein, particularly along its posterior wall. PMID- 15541133 TI - The Lahey clinic experience with continent urinary diversion. AB - OBJECTIVE: To compare the morbidity, mortality and clinical outcome of catheterizable continent urinary reservoirs (CUR) with orthotopic neobladders (ONB) at our institution. PATIENTS AND METHODS: Between September 1985 and October 2001, 238 patients (84 women and 154 men) had a continent urinary diversion, including 125 ONBs and 113 CURs. The charts of these patients were reviewed retrospectively and the patients interviewed by telephone when possible (for continence data and overall satisfaction). Over the 16 years the relative frequency of ONB diversion increased steadily and thus the mean follow-up was significantly longer for the CUR (9.4 years) than for the ONB group (5.2 years) (P < 0.001). This bias was addressed by comparing these large groups for the early outcome only. Separately, the long-term outcome was analysed, comparing the 40 most recently constructed CURs with all 113 ONBs; this gave a mean follow-up of 5.2 years for ONB and 5.9 years for CUR (not significant, P = 0.23). RESULTS: Of the 238 continent diversions, 125 were ONBs and 113 CURs; most patients had a diagnosis of transitional cell carcinoma before surgery. The mean age at surgery was 59.1 years for the ONB and 54.8 years for the CUR group (P < 0.001). Men were significantly more likely than women to have had an ONB. There were two deaths after surgery in the ONB and none in the CUR group. The hospital stay was significantly longer for the CUR than for the ONB group; the likelihood of an intensive care unit stay, estimated blood loss and reoperation rate were higher in the CUR group. There was no significant difference in the rates of short-term (<30 days) complications. The analysis of the time-controlled groups showed significantly more long-term (>30 days) complications and of reoperation in the CUR group. Fifty-one patients with ONB and 19 with CURs were contacted by telephone; of those with an ONB, 43 (84%) had daytime continence (<1 pad/day) while 13 (25%) were continent at night (<1 pad). Fifteen of 19 with a CUR reported full day and night-time continence. From separate telephone interviews, overall satisfaction was high for both groups (mean 4.5, scale 0-5), expectations were met in 92% for both, and 94% in both would choose the same procedure again if confronted with the same set of circumstances. CONCLUSIONS: Both ONB and CUR offer an excellent functional outcome, as reflected by patient satisfaction and continence rates. PMID- 15541132 TI - Is povidone iodine an alternative to silver nitrate for renal pelvic instillation sclerotherapy in chyluria? AB - OBJECTIVE: To compare the efficacy and toxicity of 1% silver nitrate, 0.2% povidone iodine and 50% dextrose in renal pelvic instillation sclerotherapy (RPIS) for chyluria. PATIENTS AND METHODS: In a prospective randomized comparative study from January 1999 to June 2003, 106 patients (61 males and 45 females; mean age 36 years, sd 12, range 14-65) were randomized to receive 1% silver nitrate, 0.2% povidone iodine or 50% dextrose as RPIS. In all, nine doses were given at 8-h intervals, and patients followed at 6 weeks and then at 3 monthly intervals. Patients with 'persistence' or 'recurrence' of chyluria were treated with second course of RPIS using same sclerosant. RESULTS: The dextrose treatment was discontinued at mid-term because of poor success (one of 21 patients, P < 0.001). Of 85 patients, 44 received silver nitrate and 41 povidone iodine; both groups were well-matched and the mean follow-up was 28.4 and 23.3 months, respectively. 'Immediate clearance' was recorded in 91% and 98%, and recurrence in 21% and 22% of patients after the first course of RPIS, after silver nitrate and povidone, respectively; Kaplan-Meier estimates of 'disease free duration' in the two groups (23.6 vs 20.1 months) were also similar (P = 0.7906). The cumulative success rate after two courses of RPIS was 82% (silver nitrate) and 83% (povidone; P = 1.0). Five (11%) patients in the silver nitrate and one (2%) in the povidone group had significant flank pain during treatment. CONCLUSIONS: Povidone iodine 0.2% is as effective for RPIS as 1% silver nitrate. PMID- 15541134 TI - Neobladder-vaginal fistula after cystectomy and orthotopic neobladder construction. AB - OBJECTIVE: To determine the potential surgical and clinical factors that contribute to the development of neobladder-vaginal fistula (NVF) after cystectomy and orthotopic neobladder (ONB) construction in women. PATIENTS AND METHODS: Of 37 patients who had vaginal-sparing cystectomy, the records of four who developed a NVF after radical cystectomy and ONB construction were reviewed. Retrospective clinical and surgical information was collected, including patient demographics, tumour pathology, surgical technique, presenting symptoms, and method and efficacy of surgical repair. RESULTS: In two of the four patients who developed a NVF a small injury to the anterior vaginal wall was noted during surgery and closed primarily. All patients presented with severe urinary incontinence. The NVF was diagnosed after cystoscopy and/or speculum examination. Three of the four patients had an attempted surgical repair, including one obturator flap interposition, one rectus flap interposition, and one primary two layer closure. To date, one patient is fistula-free and two were subsequently converted to an ileal conduit or continent cutaneous diversion because the fistula recurred. The fourth patient developed a NVF in association with local tumour recurrence and underwent conversion to an ileal conduit. CONCLUSION: The development of a NVF is a significant complication after cystectomy. Inadvertent injury to the vaginal wall is an important predisposing factor to subsequent NVF development. The repair of a NVF is often difficult; upon diagnosis, conversion to a continent cutaneous urinary diversion may be considered. PMID- 15541135 TI - Single-layer small intestinal submucosa or tunica vaginalis flap for correcting penile chordee. AB - OBJECTIVE: To evaluate the use of single-layer small intestinal submucosa (SIS) and tunica vaginalis flap (TVF) for covering defects in the ventral surface of the tunica albuginea to correct severe penile chordee. MATERIALS AND METHODS: In all, 24 New Zealand white rabbits had a rectangular area excised from the ventral surface of tunica albuginea. In 12 rabbits TVF was used to cover the defect and in the remaining animals single-layer SIS was used. Animals were killed in groups of four at 2-, 6- and 12-week intervals after surgery. Before death in the 12 week group, an artificial erection was induced and cavernosography performed. Transverse sections of the penis at the graft site were stained with haematoxylin and eosin and Masson's trichrome, and examined microscopically. RESULTS: None of the animals developed haematoma or bleeding. The mean operative duration for TVF and SIS grafts were 56 and 29 min, respectively (P < 0.001). At the time of autopsy, there was no contracture in any of the rabbits. Of the 8 rabbits assessed, all had a straight rigid erection and cavernosography showed evidence of an intact corporal veno-occlusive mechanism. Histologically at 6 and 12 weeks, the mesothelial layers of the TVF and the SIS graft were completely replaced by well-collagenized tissue similar to that of normal tunica albuginea, with no inflammatory infiltrate. CONCLUSIONS: Both the TVF and single-layer SIS graft are viable comparable options for corporal body grafting. The 'off-the-shelf' availability, significantly quicker operation and absence of donor site morbidity make single-layer SIS better than TVF for correcting chordee. PMID- 15541136 TI - Comparative study of dartos fascia and tunica vaginalis pedicle wrap for the tubularized incised plate in primary hypospadias repair. AB - OBJECTIVES: To report a prospective comparative study on using dartos fascia, i.e. subcutaneous tissue of penile skin and tunica vaginalis pedicled wrap (TVPW) from the parietal layer of the tunica vaginalis of the testis, for a one-stage tubularized incised-plate (TIP) repair for hypospadias. PATIENTS AND METHODS: Forty-nine patients (mean age 4.6 years, range 1-22) with hypospadias of different types (varying from coronal to penoscrotal) were repaired in one of three hospitals over 3 years. All patients were repaired using the TIP technique, with dartos fascia wrap used in 20 and TVPW in 29, without using a loupe or microscope during surgery. Urinary diversion and splinting were provided by a urethral catheter. The operative duration for both groups was similar at approximately 2 h. RESULTS: In the dartos fascia group there were three (15%) fistulae, but there were none in the TVPW group. CONCLUSIONS: Although the dissection for TVPW seems to be cumbersome theoretically, it is not difficult. The combination of TIP and TVPW in primary repair may be a good alternative to other techniques. PMID- 15541137 TI - Enhanced objective quantitative cystometric analysis of compliance and contractility. AB - OBJECTIVE: To show, for pressure-time data from cystometrography (CMG), the potential practical clinical application of automatically identified, displayed, analysed and quantified compliance and contractility, as undesirable high pressure detrusor storage may be caused by inefficient compliance or uninhibited contractions (UNC). MATERIAL AND METHODS: Bladder contractility was measured by UNC and compliance by relaxed-state detrusor pressure (RSDP), i.e. the detrusor (bladder-abdominal) pressure with all UNC removed. Forty-one CMG examinations were used retrospectively to: (i) validate the separation and identification, by comparing the resulting separate graphs (data) of UNC and RSDP with an expanded time scale for raw vesical and rectal data; (ii) show that the separation is correct by examples; and (iii) show the potential practical utility by results for typical cases. RESULTS: Separation into RSDP and UNC was correctly identified and plotted. The examples showed the utility and four types of UNC ('high', contractions of >25 cmH(2)O of long duration; 'medium', >25 cmH(2)O of short duration; 'low', 4-25 cmH(2)O of short duration; and 'frequent', of 2-6 cmH(2)O). CONCLUSIONS: UNCs as small as 2 cmH(2)O can be detected and measured. The explicit enhanced estimate of compliance and contractility will be useful in the follow-up when comparing different patients and studies, and assist in more appropriate diagnosis and treatment. Because the treatment for bladders with poor contractility differs greatly from those with detrusor instability, the ability to reliably and accurately differentiate between these causes is important. PMID- 15541138 TI - cGMP-generating cells in the bladder wall: identification of distinct networks of interstitial cells. AB - OBJECTIVE: To identify cells which might contribute to the complex physiological responses of the guinea-pig bladder, and specifically to describe the distribution and types of cell in the bladder wall of the guinea pig which respond to nitric oxide (NO) with an increase in intracellular cGMP, i.e. putative interstitial cells (ICs). MATERIALS AND METHODS: The whole bladder was removed from 11 male guinea pigs killed by cervical dislocation. Sections of the bladder wall, from the dome lateral wall and base, were isolated and incubated separately in Krebs' solution at 36 degrees C, gassed with 95% O(2) and 5% CO(2), and containing 1 mmol/L of the nonspecific phosphodiesterase inhibitor isobutyl methyl-xanthene. Individual pieces of tissue were then exposed to 100 micromol/L of the NO donor NONOate for 10 min; control tissues remained in Krebs' solution. Tissues were then fixed in 4% paraformaldehyde and processed for immunohistochemistry. cGMP and neuronal NO synthase (nNOS) were subsequently visualized using appropriate primary and secondary antibodies. RESULTS: Cells responding to NO with an increase in cGMP were detected in the dome, lateral wall and base, with positive cells in the thin outer surface of the wall (muscle coat), associated with muscle bundles in an outer layer of muscle, and in a region immediately beneath the urothelium. These cells (not urothelium, smooth muscle or vascular) are described as interstitial cells. Superficial urothelial umbrella cells were apparent and were strongly cGMP-positive. A high density of interstitial cells was associated with muscle bundles on the outer aspects of the wall, while few cells were detected on inner bundles. Thus there appeared to be two distinct types of muscle, inner and outer, with no obvious orientation of the fibres in each layer. Both muscle groups contained fibres expressing nNOS. In the outer muscle layer most of these fibres co-localized with cGMP, suggesting that different populations of nerves innervate each layer. There were more nNOS positive fibres in the base of the bladder than in the dome. Three populations of cGMP-positive interstitial cells were associated with the outer muscle layer; cells in the outer surface (muscle coat interstitial cells, MC-ICs), cells on the surface of the bundles (superficial, SM-ICs) and cells within the muscle bundles (intramuscular, IM-ICs). The IM-ICs form a network in close apposition to the smooth muscle cells while the SM-ICs may connect adjacent muscle bundles and connect to the MC-ICs. Thus, there is a network linking potentially the muscle cells in the outer muscle bundles. cGMP-positive cells were also detected in the suburothelial layer (suburothelial, SU-ICs) which had a different structure to the cells associated with muscle, had a oval cell bodies with bifurcating processes and appeared to form a complex network; they were prevalent in the base and virtually absent in the dome. CONCLUSIONS: There are structures within the bladder wall that can be identified and categorized by the ability of the constituent cells to increase intracellular cGMP in response to NO; these cells have been defined as ICs. Two distinct networks were identified, one associated with the outer muscle layers and another lying immediately beneath the urothelium, predominantly in the base of the bladder. The functions of these cells and networks are unknown; their possible roles in complex motor activity, urothelial signalling and bladder pathophysiology are discussed. PMID- 15541139 TI - Involvement of 5-hydroxytryptamine (HT)7 receptors in the 5-HT excitatory effects on the rat urinary bladder. AB - OBJECTIVE: To investigate the in vitro and in vivo effects of 5-hydroxytryptamine (5-HT) on the rat urinary bladder and to characterize the receptors involved in mediating these pharmacological effects by using selective antagonists. MATERIALS AND METHODS: Female Wistar rats (250-350 g) were used for all studies. In vitro, detrusor muscle strips were mounted between two platinum electrodes in organ baths filled with a modified Krebs' solution bubbled with 95% O(2) and 5% CO(2) at 37 degrees C. After equilibration and a contraction to 80 mmol/L KCl, strips were exposed to electrical field stimulation for 30 min and incubated with the antagonist or vehicle for a further 30 min, then a 5-HT concentration-response curve (CRC) was obtained. In vivo, rats were anaesthetized with pentobarbital, and the ureters and urethra ligated, the bladder catheterized and infused with saline. 5-HT (3-100 microg/kg intravenous) dose-dependently increased intravesical pressure (IVP). After administering 5-HT at 30 microg/kg three times at 10 min intervals (controls), one dose of antagonist was perfused for 5 min and, after a further 5 min, 30 microg/kg 5-HT was tested again. This cycle was repeated four times using increasing doses of the antagonist to be tested. RESULTS: In vitro, 5-HT (0.01-100 micromol/L) induced a concentration-dependent enhancement of the neurogenic response, with a mean (sd) pEC(50) of 6.36 (0.15) and E(max) of 41.1 (4.6)% KCl (eight rats). In unstimulated tissues, 5-HT induced no contractile effect. Selective 5-HT(4), 5-HT(3) and 5-HT(1A) receptor antagonists had no effect on the 5-HT potentiating effects. The potentiating effect of 5-HT was antagonized by mesulergine at 0.3 micromol/L, R(+)lisuride at 0.3 micromol/L and the selective 5-HT(7) receptor antagonist SB-258741 at 0.3 micromol/L. In vivo, in anaesthetized rats, IVP increases induced by repeated doses of 30 microg/kg 5-HT were reproducible. R(+)lisuride (3-100 microg/kg) dose dependently inhibited the 5-HT-induced increase of IVP. At the maximum dose tested, R(+)lisuride almost totally inhibited the 5-HT effect. CONCLUSIONS: In rat isolated detrusor muscle the 5-HT(7) receptor antagonists SB-258741, R(+)lisuride and mesulergine blocked the 5-HT potentiating effect with the expected potency. Moreover, in anaesthetized rats, R(+)lisuride abolished 5-HT effects on IVP at doses that antagonize physiological effects known to be mediated by 5-HT(7) receptor activation in several animal species. These results suggest the involvement of 5-HT(7) receptors in the modulation of rat bladder contraction both in vitro and in vivo. PMID- 15541140 TI - Urethral replacement: a comparison between small intestinal submucosa grafts and spontaneous regeneration. AB - OBJECTIVE: To evaluate the use of commercially available single-layer small intestinal submucosa (SIS) for urethral replacement, both as an onlay and as a tube, in a rabbit model. MATERIALS AND METHODS: Thirty-six male rabbits were assigned to four experimental groups. Group 1 had the ventral wall of the penile urethra excised for 15 mm; in group 2 this created defect was patched with a SIS onlay graft; group 3 had complete excision of a 15 mm segment of the penile urethra; and in group 4, this created defect was replaced with a SIS tube graft. In all rabbits the urethra was stented for 2 weeks. A retrograde urethrogram was taken in all rabbits before death at 3, 6 and 12 weeks after surgery. The urethra was then exposed, examined carefully and excised for histopathological examination. RESULTS: In groups 1 and 2 the retrograde urethrograms were normal in 13 rabbits and there was relative narrowing in two rabbits in group 1 and three in group 2. In groups 3 and 4 all rabbits developed urethral fistulae or strictures. Histological examination of the urethra showed epithelial regeneration supported by smooth muscle backing in all rabbits in group 1, while rabbits in group 2 showed no regeneration of smooth muscle. By contrast, rabbits in groups 3 and 4 showed incomplete regeneration and progressive fibrosis. CONCLUSIONS: Single-layer SIS is not a suitable urethral substitute in this animal model. When used as an onlay, healing is inferior to spontaneous urethral regeneration, as SIS impedes smooth muscle cell regeneration. When used as a tube, there is complete scarring and urethral luminal occlusion. PMID- 15541141 TI - The brain leads the way. PMID- 15541142 TI - Use of a deflectable laser fibre in a flexible cystoscope (nephroscope). PMID- 15541143 TI - Treatment of transurethral resection syndrome with intravenous 29.2% saline. PMID- 15541144 TI - Vaginal construction using sigmoid colon in children and young adults. PMID- 15541145 TI - MTOPS: conclusions about invasive therapy for BPH should be interpreted with caution. PMID- 15541146 TI - BJU International June issue DVD. PMID- 15541148 TI - Consensus statement on urethral trauma. PMID- 15541149 TI - 'Vanishing' prostate cancer in radical prostatectomy specimens: incidence and long-term follow-up in 38 cases. PMID- 15541150 TI - Increasing prostate biopsy cores based on volume vs the sextant biopsy: a prospective randomized controlled clinical study on cancer detection rates and morbidity. PMID- 15541151 TI - Quality of life compared during pharmacological treatments and clinical monitoring for non-localized prostate cancer: a randomized controlled trial. PMID- 15541152 TI - A single relaxing incision to correct different types of penile curvature: surgical technique based on geometrical principles. PMID- 15541154 TI - Strengthening the prevention of HIV/AIDS-related oral disease: a global approach. PMID- 15541155 TI - Recurrent herpes labialis in US children and youth. AB - OBJECTIVE: This study reports data from the Third National Health and Nutrition Examination Study, 1988-1994 (NHANES III). METHODS: NHANES III was a complex, multistage sample of 33 994 civilian, non-institutional individuals from 19 528 households. Dentist examiners were trained to recognize, classify oral mucosal lesions to include recurrent herpes labialis (RHL). Subjects >or=8 years of age were asked if they had cold sores in the past year and serologic tests for herpes virus type 1 (HSV-1) and type 2 (HSV-2) were performed on blood of youth >12 years of age. RESULTS: Examinations were performed on 10 032 individuals 2-17 years of age. Overall point prevalence was 1.42% (0.69-2.15); annual prevalence in individuals 8-17 years of age was 14.77% (12.74-16.80); and serologic prevalence of HSV-1 in youth 12-17 years of age was 43.18% (38.88-47.48). When the data were subset to youth 12-17, annual prevalence for seropositives was 24.13% (20.44-27.82) compared with 16.87 (14.16-19.57) for all subjects. Approximately 25% of the seropositive youth had at least one recurrence in the past year. CONCLUSION: As RHL is a recurrent infection, prevalence in a population will be related to the proportion of the population that has been infected with herpes simplex virus. When lesion-specific prevalences are cited in the literature, they should be stratified by covariates known to be associated with them. Future studies should examine RHL prevalence in infected individuals. PMID- 15541156 TI - A comparison of a computer-based questionnaire and personal interviews in determining oral health-related behaviours. AB - BACKGROUND: A major challenge for survey-based research is to reduce bias. In an interview, subjects may claim more favourable behaviour to please the interviewer or comply with accepted norms. OBJECTIVES: This study aimed to: (i) determine if adolescents give different answers when using a self-completed computer administered oral health questionnaire compared with a personal interview, (ii) ascertain if responses to a face-to-face interview are dependant on whether a dentist or nonclinical researcher administered the questionnaire and (iii) examine if responses were influenced by whether they undertook the computer questionnaire first or second. METHODS: A randomized crossover design was used to investigate the responses to 15 closed questions on oral health-related practices. These were administered to 453, 12-year olds attending school dental inspections in South Wales. RESULTS: A total of 449 valid pairings of computer/interview responses were available for analysis. Responses to the questionnaire variables demonstrated good to very good levels of agreement (kappa 0.68-0.90) when comparing the face-to-face and computer-administered questionnaire. With the exception of questions on dental attendance, responses were not influenced by whether the questions were posed by the research officer or the dentist. A minimal order effect could be detected when undertaking the computer-administered questionnaire first. CONCLUSIONS: Whilst a study such as this cannot determine the true validity of the responses achieved, it is concluded that a computer-administered questionnaire, comprising closed questions, can be utilized to determine oral health-related behaviours in oral health surveys. PMID- 15541157 TI - Assessment of different methods for diagnosing dental caries in epidemiological surveys. AB - AIMS: The aims of the study were: (i) to assess different clinical diagnostic methods of dental caries during epidemiological surveys; (ii) to determine which combinations of methods and diagnostic adjuncts show the best performances in epidemiological surveys when compared with examinations performed in a traditional dental setting (standard); (iii) to evaluate the influence of including noncavitated (NC) lesions in dental caries estimation. METHODS: Forty 12-year-old children were divided into low and moderate caries prevalence groups. The individuals were submitted to 12 epidemiological examinations (in an outdoor setting), which combined three methods (blade, mirror and mirror + CPI (Community Periodontal Index) dental probe) with or without diagnostic adjuncts (previous dental brushing and dental drying). The last examination was performed in a traditional dental setting (standard examination). The unit of measure was the DMFS (decayed, missing and filled surfaces) index according to WHO criteria. The variance analysis, Dunnet's and Tukey's tests were applied. RESULTS: For the DMFS analysis, the visual/tactile method, with or without diagnostic adjuncts, was the best method for both groups, presenting a performance higher than 90% when compared with the standard examination, except for the examinations without previous dental brushing for the low caries prevalence group. Previous dental brushing was more relevant than dental drying (P = 0.0054). All of the epidemiological examinations underestimated the NC diagnosis even with the association of diagnostic adjuncts when compared with the standard examination. CONCLUSION: The visual-tactile (for both groups) and the visual (mirror) methods plus dental brushing (for the moderate group) are appropriate for diagnosing cavitated lesions, but not NC lesions. PMID- 15541158 TI - Temporal stability of the theory of planned behavior: a prospective analysis of sugar consumption among Ugandan adolescents. AB - PURPOSE: This study addressed three questions: What is the power of the Theory of Planned Behavior (TPB) in predicting adolescents' intended and self-perceived consumption of non-milk extrinsic sugars using a non-intervention prospective approach? To what extent do the TPB constructs change across time following adolescents' mere exposure to an oral health survey? Do changes in self-perceived sugar consumption at follow-up associate with changes in behavioral intention as predicted by the TPB? METHOD: A survey was conducted in Kampala (urban) and Lira (rural) and 1146 secondary school students completed questionnaires assessing the TPB at school (Time 1). A random sub-sample of 415 students was selected from the original survey of which 372 students were examined clinically. After 3 months (Time 2), the questionnaire was administered a second time in the sub-sample. All analyses are based on the number of students who participated on both survey occasions, n = 372. RESULTS: Attitudes and perceived behavioral control predicted intended sugar consumption at Time 1 and Time 2, accounting for 58% (DeltaR(2) = 0.58) and 19% (DeltaR(2) = 0.19) of the variance, respectively. Time 1 intention provided significant prediction of Time 2 self-perceived sugar consumption with DeltaR(2) = 0.5. Adolescents with high-caries experience more than their counterparts with low, changed towards weaker intentions and less frequent sugar consumption across the survey period. Mean sugar consumption scores changed from 2.6 to 2.7 (ns), 3.1 to 2.6 (P < 0.001) and 2.3 to 3.2 (P < 0.001) among adolescents who, respectively, remained stable, increased and decreased their intentions across time. CONCLUSION: This study supports the validity of the TPB in predicting intended and self-perceived sugar consumption prospectively. PMID- 15541159 TI - Relative effects of pre- and post-eruption water fluoride on caries experience by surface type of permanent first molars. AB - OBJECTIVES: To determine the relative pre- and post-eruption exposure effects of fluoridated water on the caries experience of different surface types of first permanent molars. METHODS: Parental questionnaires covering residential history of participants were linked to the oral examinations of 6-15-year-old Australian children conducted in 1992 by the School Dental Services of South Australia and Queensland. Percentage of lifetime exposed to optimally fluoridated water pre- (PRE) and post-eruption (POST) was calculated with respect to tooth eruption age. Combined pre- and post-eruption categories were created to test PRE against POST exposure: PRE and POST = 0, PRE < POST, PRE = POST and in the range 0-90% of lifetime exposure, PRE > POST and, PRE and POST >or= 90% lifetime exposure. These categories were used as indicator variables in linear regression models with PRE and POST = 0 as reference in an analysis of first permanent molar DMFS scores overall and by surface type. RESULTS: Participation rates were 69.7% in South Australia (n = 9690) and 55.6% in Queensland (n = 10 195). Compared with the reference, the categories PRE > POST (beta = -0.033), PRE = POST (beta = -0.028) in the range 0-90% and, PRE and POST >or= 90% (beta = -0.055) showed significantly lower caries overall (P < 0.01), with a similar pattern for pit and fissure surface caries (beta = -0.035, -0.031 and -0.052, respectively). Only a high PRE and POST exposure decreased caries levels significantly in the approximal (beta = -0.038; P < 0.01) and free smooth surfaces (beta = -0.023; P = 0.03). CONCLUSIONS: Pre-eruption exposure was important for a caries preventive effect on first permanent molars in children 6-15 years old since post-eruption exposure alone could not lower caries levels significantly. For pit and fissure surfaces, a high pre-eruption exposure could decrease caries levels significantly. However, for other surface types, only a high pre- and post eruption exposure produced a caries preventive effect. PMID- 15541160 TI - Influence of perceived provider performance on satisfaction with oral health care among adolescents. AB - OBJECTIVE: To investigate satisfaction with dental care received at the last dental appointment among adolescents and to analyse factors influencing their satisfaction scores. METHOD: A total of 1146 subjects (mean age 15.8 years) attending secondary schools in Kampala (urban, n = 591) and Lira (rural, n = 555) completed structured questionnaires at school in 2001. RESULTS: A total of 63% and 75% of Kampala and Lira students, respectively, reported attendance to dental clinics during the previous 2 years. The corresponding rates of students who confirmed satisfaction with oral health care services received were 73 and 77. In a logistic regression model, the students of Kampala who attended a dentist more than once, had no painful experience at the visit, evaluated their oral condition positively, were satisfied with the dentist's communication and dentist's information, were more likely to be satisfied with the oral health services received (OR = 1.7, 2.2, 4.1, 2.9 and 4.9, respectively). Regarding rural students, being satisfied with oral condition, dentist's communication and dentist's information were associated with higher odds of being satisfied with oral health care services (OR = 2.9, 1.9 and 2.3, respectively). CONCLUSION: Inter-personal interaction with the dentist is a key determinant in establishing satisfaction with dental care among urban as well as rural adolescents. PMID- 15541161 TI - Children's coping with pain during dental care. AB - OBJECTIVES: The purpose of this study was (i) to assess the coping strategies of 11-year-old children when dealing with pain at the dentist, (ii) to determine the extent to which the level of the children's dental fear and their experience with pain at the dentist are related to their ability to cope and their choice of strategies, and (iii) to analyse the possible differences between subsamples concerning dental caries. METHODS: The coping strategies were investigated using the Dental Cope Questionnaire (n = 597); the level of dental fear was assessed using the Children's Fear Survey Schedule (CFSS-DS); a question is asked whether a child had experienced pain at the dentist in the past and dental caries was assessed using the DMFS index. RESULTS: The results show that 11-year olds use a variety of coping strategies. Internal strategies are used most frequently, external coping strategies are used less frequently, and destructive strategies are hardly used. The subjects rate internal and external strategies as effective. Children with pain experience and fearful children use more coping strategies, with fearful children using more internal strategies. Reported pain and anxiety were related to the dental status. CONCLUSIONS: The use and choice of coping strategies seems to be at least partly determined by the level of dental fear and the child's experience with pain. PMID- 15541162 TI - Salivary fluoride levels following application of fluoride varnish or fluoride rinse. AB - OBJECTIVES: This study examined the concentration of fluoride in whole saliva over time following the application of a fluoride varnish or a single rinse with a fluoride solution. METHODS: A two-period, two-treatment randomized cross-over experimental trial with a 2-week washout period was used with 16 adult subjects. In the first period, eight subjects rinsed once with a 0.05% NaF solution and 8 subjects had 5.0% NaF varnish applied to facial and lingual surfaces of 20 teeth. Stimulated whole saliva was collected at baseline, 5 and 15 min, 1, 2, 4, 8, 12, 24, 32, 48, 56, 72, 80, 96, 104 h. After the washout period each subject was switched to the other treatment and saliva was collected at the same intervals. Salivary fluoride content was measured with the micro-diffusion method. RESULTS: The NaF levels peaked at 5 min after application for both varnish (mean +/- SE 24.5 +/- 5.0 ppm) and rinse (3.2 +/- 0.8 ppm). Mean NaF levels returned to baseline, on average, within 2 h for the rinse and within 24 h for the varnish. The maximum fluoride levels were significantly greater (P < 0.01) with the varnish than with the rinse and remained above baseline levels for a longer duration. CONCLUSIONS: Salivary fluoride levels with the rinse returned to baseline, on average, in 2 h while they remained elevated for, on average, 24 h with the varnish. Salivary fluoride levels from the varnish were found to be comparable with those in previous studies for 1.1% neutral NaF. PMID- 15541164 TI - Modeling effective treatment of psychoses in the elderly: a focus on avoiding movement disorders. PMID- 15541165 TI - Caring for the whole person: integrated health care for older adults with severe mental illness and medical comorbidity. PMID- 15541166 TI - Pharmacology of antipsychotics in the elderly: a focus on atypicals. PMID- 15541167 TI - Neuroleptic-induced movement disorders: deconstructing extrapyramidal symptoms. PMID- 15541168 TI - Optimizing atypical antipsychotic treatment strategies in the elderly. PMID- 15541169 TI - A trial design for evaluation of empiric programming of implantable cardioverter defibrillators to improve patient management. AB - The delivery of implantable cardioverter defibrillator (ICD) therapy is sophisticated and requires the programming of over 100 settings. Physicians tailor these settings with the intention of optimizing ICD therapeutic efficacy, but the usefulness of this approach has not been studied and is unknown. Empiric programming of settings such as anti-tachycardia pacing (ATP) has been demonstrated to be effective, but an empiric approach to programming all VT/VF detection and therapy settings has not been studied. A single standardized empiric programming regimen was developed based on key strategies with the intention of restricting shock delivery to circumstances when it is the only effective and appropriate therapy. The EMPIRIC trial is a worldwide, multi center, prospective, one-to-one randomized comparison of empiric to physician tailored programming for VT/VF detection and therapy in a broad group of about 900 dual chamber ICD patients. The trial will provide a better understanding of how particular programming strategies impact the quantity of shocks delivered and facilitate optimization of complex ICD programming. PMID- 15541170 TI - Calibration and assessment of channel-specific biases in microarray data with extended dynamical range. AB - BACKGROUND: Non-linearities in observed log-ratios of gene expressions, also known as intensity dependent log-ratios, can often be accounted for by global biases in the two channels being compared. Any step in a microarray process may introduce such offsets and in this article we study the biases introduced by the microarray scanner and the image analysis software. RESULTS: By scanning the same spotted oligonucleotide microarray at different photomultiplier tube (PMT) gains, we have identified a channel-specific bias present in two-channel microarray data. For the scanners analyzed it was in the range of 15-25 (out of 65,535). The observed bias was very stable between subsequent scans of the same array although the PMT gain was greatly adjusted. This indicates that the bias does not originate from a step preceding the scanner detector parts. The bias varies slightly between arrays. When comparing estimates based on data from the same array, but from different scanners, we have found that different scanners introduce different amounts of bias. So do various image analysis methods. We propose a scanning protocol and a constrained affine model that allows us to identify and estimate the bias in each channel. Backward transformation removes the bias and brings the channels to the same scale. The result is that systematic effects such as intensity dependent log-ratios are removed, but also that signal densities become much more similar. The average scan, which has a larger dynamical range and greater signal-to-noise ratio than individual scans, can then be obtained. CONCLUSIONS: The study shows that microarray scanners may introduce a significant bias in each channel. Such biases have to be calibrated for, otherwise systematic effects such as intensity dependent log-ratios will be observed. The proposed scanning protocol and calibration method is simple to use and is useful for evaluating scanner biases or for obtaining calibrated measurements with extended dynamical range and better precision. The cross platform R package aroma, which implements all described methods, is available for free from http://www.maths.lth.se/bioinformatics/. PMID- 15541171 TI - Functional interdependence between septin and actin cytoskeleton. AB - BACKGROUND: Septin2 is a member of a highly conserved GTPase family found in fungi and animals. Septins have been implicated in a diversity of cellular processes including cytokinesis, formation of diffusion barriers and vesicle trafficking. Septin2 partially co-localises with actin bundles in mammalian interphase cells and Septin2-filamentmorphology depends upon an intact actin cytoskeleton. How this interaction is regulated is not known. Moreover, evidence that Septin2 is remodelled or redistributed in response to other changes in actin organisation is lacking. RESULTS: Septin2 filaments are associated with actin fibres, but Septin2 is not associated with actin at the leading edge of moving cells or in ruffles where actin is highly dynamic. Rather, Septin2 is spatially segregated from these active areas and forms O- and C-shaped structures, similar to those previously observed after latrunculin treatment. FRAP experiments showed that all assemblies formed by Septin2 are highly dynamic with a constant exchange of Septin2 in and out of these structures, and that this property is independent of actin. A combination of RNAi experiments and expression of truncated forms of Septin2 showed that Septin2 plays a significant role in stabilising or maintaining actin bundles. CONCLUSION: We show that Septin2 can form dynamic structures with differing morphologies in living cells, and that these morphologies are dependent on the functional state of the actin cytoskeleton. Our data provide a link between the different morphological states of Septin2 and functions of Septin2 in actin-dynamics, and are consistent with the model proposed by Kinoshita and colleagues, that Septin2 filaments play a role in stabilisation of actin stress fibres thus preventing actin turnover. PMID- 15541172 TI - Identification of a panel of tumor-associated antigens from breast carcinoma cell lines, solid tumors and testis cDNA libraries displayed on lambda phage. AB - BACKGROUND: Tumor-associated antigens recognized by humoral effectors of the immune system are a very attractive target for human cancer diagnostics and therapy. Recent advances in molecular techniques have led to molecular definition of immunogenic tumor proteins based on their reactivity with autologous patient sera (SEREX). METHODS: Several high complexity phage-displayed cDNA libraries from breast carcinomas, human testis and breast carcinoma cell lines MCF-7, MDA MB-468 were constructed. The cDNAs were expressed in the libraries as fusion to bacteriophage lambda protein D. Lambda-displayed libraries were efficiently screened with sera from patients with breast cancer. RESULTS: A panel of 21 clones representing 18 different antigens, including eight proteins of unknown function, was identified. Three of these antigens (T7-1, T11-3 and T11-9) were found to be overexpressed in tumors as compared to normal breast. A serological analysis of the 21 different antigens revealed a strong cancer-related profile for at least five clones (T6-2, T6-7, T7-1, T9-21 and T9-27). CONCLUSIONS: Preliminary results indicate that patient serum reactivity against five of the antigens is associated with tumor disease. The novel T7-1 antigen, which is overexpressed in breast tumors and recognized specifically by breast cancer patient sera, is potentially useful in cancer diagnosis. PMID- 15541173 TI - Use of interrupter technique in assessment of bronchial responsiveness in normal subjects. AB - BACKGROUND: A number of subjects, especially the very young and the elderly, are unable to cooperate and to perform forced expiratory manoeuvres in the evaluation of bronchial hyperresponsiveness (BHR). The objective of our study was to investigate the use of the interrupter technique as a method to measure the response to provocation and to compare it with the conventional PD20 FEV1. METHODS: We studied 170 normal subjects, 100 male and 70 female (mean +/- SD age, 38 +/- 8.5 and 35 +/- 7.5 years, respectively), non-smoking from healthy families. These subjects had no respiratory symptoms, rhinitis or atopic history. A dosimetric cumulative inhalation of methacholine was used and the response was measured by the dose which increases baseline end interruption resistance by 100% (PD100Rint, EI) as well as by percent dose response ratio (DRR). RESULTS: BHR at a cut-off level of 0.8 mg methacholine exhibited 31 (18%) of the subjects (specificity 81.2%), 21 male and 10 female, while 3% showed a response in the asthmatic range. The method was reproducible and showed good correlation with PD20FEV1 (r = 0.76, p < 0.005), with relatively narrow limits of agreement at 1.39 mumol and 1.27 mumol methacholine, respectively, but the interrupter methodology proved more sensitive than FEV1 in terms of reactivity (DRR). CONCLUSIONS: Interrupter methodology is clinically useful and may be used to evaluate bronchial responsiveness in normal subjects and in situations when forced expirations cannot be performed. PMID- 15541174 TI - Weather-based prediction of Plasmodium falciparum malaria in epidemic-prone regions of Ethiopia I. Patterns of lagged weather effects reflect biological mechanisms. AB - BACKGROUND: Malaria epidemics due to Plasmodium falciparum are reported frequently in the East African highlands with high case fatality rates. There have been formal attempts to predict epidemics by the use of climatic variables that are predictors of transmission potential. However, little consensus has emerged about the relative importance and predictive value of different factors. Understanding the reasons for variation is crucial to determining specific and important indicators for epidemic prediction. The impact of temperature on the duration of a mosquito's life cycle and the sporogonic phase of the parasite could explain the inconsistent findings. METHODS: Daily average number of cases was modeled using a robust Poisson regression with rainfall, minimum temperature and maximum temperatures as explanatory variables in a polynomial distributed lag model in 10 districts of Ethiopia. To improve reliability and generalizability within similar climatic conditions, we grouped the districts into two climatic zones, hot and cold. RESULTS: In cold districts, rainfall was associated with a delayed increase in malaria cases, while the association in the hot districts occurred at relatively shorter lags. In cold districts, minimum temperature was associated with malaria cases with a delayed effect. In hot districts, the effect of minimum temperature was non-significant at most lags, and much of its contribution was relatively immediate. CONCLUSIONS: The interaction between climatic factors and their biological influence on mosquito and parasite life cycle is a key factor in the association between weather and malaria. These factors should be considered in the development of malaria early warning system. PMID- 15541175 TI - Antigenized antibodies expressing Vbeta8.2 TCR peptides immunize against rat experimental allergic encephalomyelitis. AB - BACKGROUND: Immunity against the T cell receptor (TCR) is considered to play a central role in the regulation of experimental allergic encephalomyelitis (EAE), a model system of autoimmune disease characterized by a restricted usage of TCR genes. Methods of specific vaccination against the TCR of pathogenetic T cells have included attenuated T cells and synthetic peptides from the sequence of the TCR. These approaches have led to the concept that anti-idiotypic immunity against antigenic sites of the TCR, which are a key regulatory element in this disease. METHODS: The present study in the Lewis rat used a conventional idiotypic immunization based on antigenized antibodies expressing selected peptide sequences of the Vbeta8.2 TCR (93ASSDSSNTE101 and 39DMGHGLRLIHYSYDVNSTEKG59). RESULTS: The study demonstrates that vaccination with antigenized antibodies markedly attenuates, and in some instances, prevents clinical EAE induced with the encephalitogenic peptide 68GSLPQKSQRSQDENPVVHF88 in complete Freunds' adjuvant (CFA). Antigenized antibodies induced an anti idiotypic response against the Vbeta8.2 TCR, which was detected by ELISA and flowcytometry. No evidence was obtained of a T cell response against the corresponding Vbeta8.2 TCR peptides. CONCLUSIONS: The results indicate that antigenized antibodies expressing conformationally-constrained TCR peptides are a simple means to induce humoral anti-idiotypic immunity against the TCR and to vaccinate against EAE. The study also suggests the possibility to target idiotypic determinants of TCR borne on pathogenetic T cells to vaccinate against disease. PMID- 15541176 TI - Inconsistent self-reported mammography history: findings from the National Population Health Survey longitudinal cohort. AB - BACKGROUND: Self-reported information has commonly been used to monitor mammography utilization across populations and time periods. However, longitudinal investigations regarding the prevalence and determinants of inconsistent responses over time and the impact of such responses on population screening estimates are lacking. METHODS: Based on longitudinal panel data for a representative cohort of Canadian women aged 40+ years (n = 3,537) assessed in the 1994-95 (baseline) and 1996-97 (follow-up) National Population Health Survey (NPHS), we examined the prevalence of inconsistent self-reports of mammography utilization. Logistic regression models were used to estimate the associations between women's baseline sociodemographic and health characteristics and 2 types of inconsistent responses: (i) baseline reports of ever use which were subsequently contradicted by follow-up reports of never use; and (ii) baseline reports of never use which were contradicted by follow-up reports of use prior to 1994-95. RESULTS: Among women who reported having a mammogram at baseline, 5.9% (95% confidence interval (CI): 4.6-7.3%) reported at follow-up that they had never had one. Multivariate logistic regression analyses showed that women with such inconsistent responses were more often outside target age groups, from low income households and less likely to report hormone replacement therapy and Pap smear use. Among women reporting never use at baseline and ever use at follow-up, 17.4% (95%CI: 11.7-23.1%) reported their most recent mammogram as occurring prior to 1994-95 (baseline) and such responses were more common among women aged 70+ years and those in poorer health. CONCLUSIONS: Women with inconsistent responses of type (i), i.e., ever users at baseline but never users at follow-up, appeared to exhibit characteristics typical of never users of mammography screening. Although limited by sample size, our preliminary analyses suggest that type (ii) responses are more likely to be the result of recall bias due to competing morbidity and age. Inconsistent responses, if removed from the analyses, may be a greater source of loss to follow-up than deaths/institutionalization or item non response. PMID- 15541177 TI - Enhanced Dupuytren's disease fibroblast populated collagen lattice contraction is independent of endogenous active TGF-beta2. AB - BACKGROUND: Dupuytren's disease (DD) is a debilitating fibro-proliferative disorder of the hand characterized by the appearance of fibrotic lesions (nodules and cords) leading to flexion contractures of the fingers and loss of hand function. Although the molecular mechanism of DD is unknown, it has been suggested that transforming growth factor-beta2 (TGF-beta2) may play an important role in the underlying patho-physiology of the disease. The purpose of this study was to further explore this hypothesis by examining the effects of TGF-beta2 on primary cell cultures derived from patient-matched disease and normal palmar fascia tissue using a three-dimensional collagen contraction assay. METHODS: Fibroblast-populated collagen lattice (FPCL) contraction assays using primary cell cultures derived from diseased and control fascia of the same DD patients were studied in response to exogenous TGF-beta2 and neutralizing anti-TGF-beta2 antibodies. RESULTS: Contraction of the FPCLs occurred significantly faster and to a greater extent in disease cells compared to control cells. The addition of TGF-beta2 enhanced the rate and degree of collagen contraction in a dose dependent fashion for both control and diseased cells. Neutralizing anti-TGF beta2 antibodies abolished exogenous TGF-beta2 stimulated collagen contraction, but did not inhibit the enhanced basal collagen contraction activity of disease FPCL cultures. CONCLUSIONS: Although exogenous TGF-beta2 stimulated both disease and control FPCL contraction, neutralizing anti-TGF-beta2 antibodies did not affect the elevated basal collagen contraction activity of disease FPCLs, suggesting that the differences in the collagen contraction activity of control and disease FPCL cultures are not due to differences in the levels of endogenous TGF-beta2 activity. PMID- 15541178 TI - Knowledge of malaria influences the use of insecticide treated nets but not intermittent presumptive treatment by pregnant women in Tanzania. AB - BACKGROUND: To reduce the intolerable burden of malaria in pregnancy, the Ministry of Health in Tanzania has recently adopted a policy of intermittent presumptive treatment for pregnant women using sulphadoxine-pyrimethamine (IPTp SP). In addition, there is strong national commitment to increase distribution of insecticide treated nets (ITNs) among pregnant women. This study explores the determinants of uptake for both ITNs and IPTp-SP by pregnant women and the role that individual knowledge and socio-economic status has to play for each. METHODS: 293 women were recruited post-partum at Kibaha District Hospital on the East African coast. The haemoglobin level of each woman was measured and a questionnaire administered. RESULTS: Use of both interventions was associated with a reduced risk of severe anaemia (Hb<8 g/dL) compared to women who had used neither intervention (OR 0.31, 95% CI 0.14-0.67). In a logistic regression model it was found that attendance at MCH health education sessions was the only factor that predicted IPTp-SP use (OR 1.8, 95% CI 1.1-2.9) while high knowledge of malaria predicted use of ITNs (OR 2.3, 95% CI 1.1-4.9). CONCLUSION: Individual knowledge of malaria was an important factor for ITN uptake, but not for IPTp-SP use, which was reliant on delivery of information by MCH systems. When both these interventions were used, severe anaemia postpartum was reduced by 69% compared to use of neither, thus providing evidence of effectiveness of these interventions when used in combination. PMID- 15541180 TI - Factors involved in nurses' responses to burnout: a grounded theory study. AB - BACKGROUND: Intense and long-standing problems in burn centers in Tehran have led nurses to burnout. This phenomenon has provoked serious responses and has put the nurses, patients and the organization under pressure. The challenge for managers and nurse executives is to understand the factors which would reduce or increase the nurses' responses to burnout and develop delivery systems that promote positive adaptation and facilitate quality care. This study, as a part of more extensive research, aims to explore and describe the nurses' perceptions of the factors affecting their responses to burnout. METHODS: Grounded theory was used as the method. Thirty- eight participants were recruited. Data were generated by unstructured interviews and 21 sessions of participant observations. Constant comparison was used for data analysis. RESULTS: Nurses' and patients' personal characteristics and social support influenced nurses' responses to burnout. Personal characteristics of the nurses and patients, especially when interacting, had a more powerful effect. They altered emotional, attitudinal, behavioral and organizational responses to burnout and determined the kind of caring behavior. Social support had a palliative effect and altered emotional responses and some aspects of attitudinal responses. CONCLUSIONS: The powerful effect of positive personal characteristics and its sensitivity to long standing and intense organizational pressures suggests approaches to executing stress reduction programs and refreshing the nurses' morale by giving more importance to ethical aspects of caring. Moreover, regarding palliative effect of social support and its importance for the nurses' wellbeing, nurse executives are responsible for promoting a work environment that supports nurses and motivates them. PMID- 15541179 TI - Healthcare professionals' perceptions of pain in infants at risk for neurological impairment. AB - BACKGROUND: To determine whether healthcare professionals perceive the pain of infants differently due to their understanding of that infant's level of risk for neurological impairment. METHOD: Neonatal Intensive Care Units (NICU's) at two tertiary pediatric centers. Ninety-five healthcare professionals who practice in the NICU (50 nurses, 19 physicians, 17 respiratory therapists, 9 other) participated. They rated the pain (0-10 scale and 0-6 Faces Pain Scale), distress (0-10), effectiveness of cuddling to relieve pain (0-10) and time to calm without intervention (seconds) for nine video clips of neonates receiving a heel stick. Prior to each rating, they were provided with descriptions that suggested the infant had mild, moderate or severe risk for neurological impairment. Ratings were examined as a function of the level of risk described. RESULTS: Professionals' ratings of pain, distress, and time to calm did not vary significantly with level of risk, but ratings of the effectiveness of cuddling were significantly lower as risk increased [F (2,93) = 4.4, p = .02]. No differences in ratings were found due to participants' age, gender or site of study. Physicians' ratings were significantly lower than nurses' across ratings. CONCLUSION: Professionals provided with visual information regarding an infants' pain during a procedure did not display the belief that infants' level of risk for neurological impairment affected their pain experience. Professionals' estimates of the effectiveness of a nonpharmacological intervention did differ due to level of risk. PMID- 15541181 TI - Validity of the AusTOM scales: a comparison of the AusTOMs and EuroQol-5D. AB - BACKGROUND: Clinicians require brief outcome measures in their busy daily practice to document global client outcomes. Based on the UK Therapy Outcome Measure, the Australian Therapy Outcome Measures were designed to capture global therapy outcomes of occupational therapy, physiotherapy and speech pathology in the Australian clinical context. The aim of this study was to investigate the construct (convergent) validity of the Australian Therapy Outcome Measures (AusTOMs) by comparing it with the EuroQuol-5D (EQ-5D). METHODS: The research was a prospective, longitudinal cohort study, with data collected over a seven month time period. The study was conducted at a total of 13 metropolitan and rural health-care sites including acute, sub-acute and community facilities. Two hundred and five clients were asked to score themselves on the EQ-5D, and the same clients were scored by approximately 115 therapists (physiotherapists, speech pathologists and occupational therapists) using the AusTOMs at admission and discharge. Clients were consecutive admissions who agreed to participate in the study. Clients of all diagnoses, aged 18 years and over (a criteria of the EQ 5D), and able to give informed consent were scored on the measures. Spearman rank order correlation coefficients were used to analyze the relationships between scores from the two tools. The clients were scored on the AusTOMs and EQ-5D. RESULTS: There were many health care areas where correlations were expected and found between scores on the AusTOMs and the EQ-5D. CONCLUSION: In the quest to measure the effectiveness of therapy services, managers, health care founders and clinicians are urgently seeking to undertake the first step by identifying tools that can measure therapy outcome. AusTOMs is one tool that can measure global client outcomes following therapy. In this study, it was found that on the whole, the AusTOMs and the EQ-5D measure similar constructs. Hence, although the validity of a tool is never 'proven', this study offers preliminary support for the construct validity of AusTOMs. PMID- 15541182 TI - G-protein inwardly rectifying potassium channel 1 (GIRK 1) gene expression correlates with tumor progression in non-small cell lung cancer. AB - BACKGROUND: G-protein inwardly rectifying potassium channel 1 (GIRK1) is thought to play a role in cell proliferation in cancer, and GIRK1 gene expression level may define a more aggressive phenotype. We detected GIRK1 expression in tissue specimens from patients with non-small cell lung cancers (NSCLCs) and assessed their clinical characteristics. METHODS: Using reverse transcription-polymerase chain reaction (RT-PCR) analyses, we quantified the expression of GIRK1 in 72 patients with NSCLCs to investigate the relationship between GIRK1 expression and clinicopathologic factors and prognosis. RESULTS: In 72 NSCLC patients, 50 (69%) samples were evaluated as having high GIRK1 gene expression, and 22 (31%) were evaluated as having low GIRK1 gene expression. GIRK1 gene expression was significantly associated with lymph node metastasis, stage (p = 0.0194 for lymph node metastasis; p = 0.0207 for stage). The overall and stage I survival rates for patients with high GIRK1 gene expressed tumors was significantly worse than for those individuals whose tumors had low GIRK1 expression (p = 0.0004 for the overall group; p = 0.0376 for stage I). CONCLUSIONS: These data indicate that GIRK1 may contribute to tumor progression and GIRK1 gene expression can serve as a useful prognostic marker in the overall and stage I NSCLCs. PMID- 15541183 TI - Missing paternal demographics: A novel indicator for identifying high risk population of adverse pregnancy outcomes. AB - BACKGROUND: One of every 6 United Status birth certificates contains no information on fathers. There might be important differences in the pregnancy outcomes between mothers with versus those without partner information. The object of this study was to assess whether and to what extent outcomes in pregnant women who did not have partner information differ from those who had. METHODS: We carried out a population-based retrospective cohort study based on the registry data in the United States for the period of 1995-1997, which was a matched multiple birth file (only twins were included in the current analysis). We divided the study subjects into three groups according to the availability of partner information: available, partly missing, and totally missing. We compared the distribution of maternal characteristics, maternal morbidity, labor and delivery complications, obstetric interventions, preterm birth, fetal growth restriction, low birth weight, congenital anomalies, fetal death, neonatal death, post-neonatal death, and neonatal morbidity among three study groups. RESULTS: There were 304466 twins included in our study. Mothers whose partner's information was partly missing and (especially) totally missing tended to be younger, of black race, unmarried, with less education, smoking cigarette during pregnancy, and with inadequate prenatal care. The rates of preterm birth, fetal growth restriction, low birth weight, Apgar score <7, fetal mortality, neonatal mortality, and post-neonatal mortality were significantly increased in mothers whose partner's information was partly or (especially) totally missing. CONCLUSIONS: Mothers whose partner's information was partly and (especially) totally missing are at higher risk of adverse pregnant outcomes, and clinicians and public health workers should be alerted to this important social factor. PMID- 15541184 TI - Large bilateral adrenal metastases in non-small cell lung cancer. AB - BACKGROUND: The adrenal gland is one of the common sites of metastasis from primary lung cancer. Adrenal metastases are usually unilateral however bilateral adrenal metastases are seen in 10% of all lung cancer patients; of these 2-3% occurs at the initial presentation of non-small cell lung cancer. Secondary tumors can disrupt the structure and function of the adrenal. This can lead to adrenal hemorrhage, which constitutes a life threatening hazard for the patient. CASE PRESENTATION: A 59-year-old male presented with persisting abdominal pain. His initial work-up revealed significant anemia, an invasive process in the right upper lobe of the lung and large masses of heterogeneous texture, with hemorrhagic and necrotic elements in both adrenal glands. A biopsy confirmed it to be a large-cell carcinoma of the lungs. The patient developed severe leukocytosis akin to the paraneoplastic syndrome and died suddenly five days after the administration of chemotherapy. CONCLUSION: Intratumoral hemorrhage is a rare but life threatening complication of adrenal metastases and should be treated as soon as it has been diagnosed. If adrenalectomy is not feasible, combination chemotherapy should be applied as in metastatic disease. For choosing the appropriate chemotherapeutic regimen it is important to accurately achieve the diagnosis. PMID- 15541185 TI - Density-dependence and within-host competition in a semelparous parasite of leaf cutting ants. AB - BACKGROUND: Parasite heterogeneity and within-host competition are thought to be important factors influencing the dynamics of host-parasite relationships. Yet, while there have been many theoretical investigations of how these factors may act, empirical data is more limited. We investigated the effects of parasite density and heterogeneity on parasite virulence and fitness using four strains of the entomopathogenic fungus, Metarhizium anisopliae var. anisopliae, and its leaf cutting ant host Acromyrmex echinatior as the model system. RESULTS: The relationship between parasite density and infection was sigmoidal, with there being an invasion threshold for an infection to occur (an Allee effect). Although spore production was positively density-dependent, parasite fitness decreased with increasing parasite density, indicating within-host scramble competition. The dynamics differed little between the four strains tested. In mixed infections of three strains the infection-growth dynamics were unaffected by parasite heterogeneity. CONCLUSIONS: The strength of within-host competition makes dispersal the best strategy for the parasite. Parasite heterogeneity may not have effected virulence or the infection dynamics either because the most virulent strain outcompeted the others, or because the interaction involved scramble competition that was impervious to parasite heterogeneity. The dynamics observed may be common for virulent parasites, such as Metarhizium, that produce aggregated transmission stages. Such parasites make useful models for investigating infection dynamics and the impact of parasite competition. PMID- 15541186 TI - Molecular and epidemiologic analysis of a county-wide outbreak caused by Salmonella enterica subsp. enterica serovar Enteritidis traced to a bakery. AB - BACKGROUND: An increase in the number of attendees due to acute gastroenteritis and fever was noted at one hospital emergency room in Taiwan over a seven-day period from July to August, 2001. Molecular and epidemiological surveys were performed to trace the possible source of infection. METHODS: An epidemiological investigation was undertaken to determine the cause of the outbreak. Stool and blood samples were collected according to standard protocols per Center for Disease Control, Taiwan. Typing of the Salmonella isolates from stool, blood, and food samples was performed with serotyping, antibiotypes, and pulsed field gel electrophoresis (PFGE) following XbaI restriction enzyme digestion. RESULTS: Comparison of the number of patients with and without acute gastroenteritis (506 and 4467, respectively) during the six weeks before the outbreak week revealed a significant increase in the number of patients during the outbreak week (162 and 942, respectively) (relative risk (RR): 1.44, 95% confidence interval (CI): 1.22 1.70, P value < 0.001). During the week of the outbreak, 34 of 162 patients with gastroenteritis were positive for Salmonella, and 28 of these 34 cases reported eating the same kind of bread. In total, 28 of 34 patients who ate this bread were positive for salmonella compared to only 6 of 128 people who did not eat this bread (RR: 17.6, 95%CI 7.9-39.0, P < 0.001). These breads were produced by the same bakery and were distributed to six different traditional Chinese markets., Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) was isolated from the stool samples of 28 of 32 individuals and from a recalled bread sample. All S. Enteritidis isolates were of the same antibiogram. PFGE typing revealed that all except two of the clinical isolates and the bread isolates were of the same DNA macrorestriction pattern. CONCLUSIONS: The egg-covered bread contaminated with S. Enteritidis was confirmed as the vehicle of infection. Alertness in the emergency room, surveillance by the microbiology laboratory, prompt and thorough investigation to trace the source of outbreaks, and institution of appropriate control measures provide effective control of community outbreaks. PMID- 15541187 TI - Evaluating legume species as alternative trap crops to chickpea for management of Helicoverpa spp. (Lepidoptera: Noctuidae) in central Queensland cotton cropping systems. AB - Mounting levels of insecticide resistance within Australian Helicoverpa spp. populations have resulted in the adoption of non-chemical IPM control practices such as trap cropping with chickpea, Cicer arietinum (L.). However, a new leaf blight disease affecting chickpea in Australia has the potential to limit its use as a trap crop. Therefore this paper evaluates the potential of a variety of winter-active legume crops for use as an alternative spring trap crop to chickpea as part of an effort to improve the area-wide management strategy for Helicoverpa spp. in central Queensland's cotton production region. The densities of Helicoverpa eggs and larvae were compared over three seasons on replicated plantings of chickpea, Cicer arietinum (L.), field pea Pisum sativum (L), vetch, Vicia sativa (L.) and faba bean, Vicia faba (L.). Of these treatments, field pea was found to harbour the highest densities of eggs. A partial life table study of the fate of eggs oviposited on field pea and chickpea suggested that large proportions of the eggs laid on field pea suffered mortality due to dislodgment from the plants after oviposition. Plantings of field pea as a replacement trap crop for chickpea under commercial conditions confirmed the high level of attractiveness of this crop to ovipositing moths. The use of field pea as a trap crop as part of an area-wide management programme for Helicoverpa spp. is discussed. PMID- 15541188 TI - Morphometric analysis of Mexican and South American populations of the Anastrepha fraterculus complex (Diptera: Tephritidae) and recognition of a distinct Mexican morphotype. AB - Discriminant function and cluster analyses were performed on 19 morphometric variables of the aculeus, wing and mesonotum to determine whether populations of Anastrepha fraterculus (Wiedemann) from different parts of Mexico could be distinguished from populations from South America. Samples were collected from seven localities across Mexico, two from Brazil, and one each from Colombia and Argentina. Results showed there were statistically significant differences between Mexican and South American populations with respect to the aculeus (tip length, length of serrated section, mean number of teeth) and wing (width of S band and connection between S- and V-bands). The degree of morphological variation observed among Mexican populations was extremely low, and as a consequence, the Mexican populations were identified as a single morphotype by discriminant analysis. The 'Andean morphotype'consisting of the Colombian population, and the 'Brazilian morphotype'consisting of the two Brazilian populations plus the single Argentinian population were also distinguished. It was concluded that the macro-geographical morphotypes from Mexico, Colombia, and Brazil plus Argentina correspond to three distinct taxonomic entities. Comparisons of results with those obtained from behavioural, karyotypic, isozyme and DNA studies suggest that sufficient evidence now exists to name a new Mexican species from within the A. fraterculus complex. This will be done in a separate publication. A provisional key to the morphotypes of A. fraterculus studied is provided. PMID- 15541189 TI - Specificity of accumulation and transmission of tomato spotted wilt virus (TSWV) in two genera, Frankliniella and Thrips (Thysanoptera: Thripidae). AB - The accumulation and transmission of tomato spotted wilt virus (TSWV) was examined in second instar larvae and adults of two thrips genera, Frankliniella and Thrips. The species tested were F. occidentalis (Pergande), F. intonsa (Trybom), T. tabaciLindeman, T. setosus Moulton, T. palmi Karny and T. hawaiiensis (Morgan). In a standard petunia leaf disc assay, the efficiencies of TSWV transmission by two species of Frankliniella were higher than those of any Thrips species in the adult stage. A triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) showed that large amounts of the TSWV nucleocapsid (N) protein were present in the ELISA-positive larvae of each species, with the exception of T. palmi. The ELISA titre of and the proportion of virus-infected individuals of the two Frankliniella species increased or did not significantly change from the larval to the adult stages, whereas those of the four Thrips species decreased significantly. These results show that the specificity of virus transmission by adult thrips is probably affected by the amount of viral N protein accumulation in the adults and that the accumulation pattern from the larval to the adult stages is in between the two genera tested in the present study. PMID- 15541190 TI - Laboratory evaluations of a wild crucifer Barbarea vulgaris as a management tool for the diamondback moth Plutella xylostella (Lepidoptera: Plutellidae). AB - The term 'dead-end trap cropping' has recently been proposed to identify a plant that is highly attractive for oviposition by an insect pest, but on which offspring of the pest cannot survive. The potential of the wild crucifer Barbarea vulgaris R. Br. to allure and serve as a dead-end trap crop for the diamondback moth Plutella xylostella (L.), an important pest of cruciferous crops worldwide, was examined in laboratory experiments. When P. xylostella adults were provided with a dual-choice of plants of B. vulgaris, and Chinese cabbage Brassica campestris (L.), in one arena, adult moths laid 2.5-6.8 times more eggs on the former than on the latter. When P. xylostella adults were provided with a dual choice of plants of B. vulgaris and common cabbage Brassica oleracea L., adult moths laid virtually all their eggs on the former and ignored the latter. Nearly all P. xylostella eggs laid on the three species of plants hatched successfully, but nearly all individuals on plants of B. vulgaris died as neonates or early instar larvae, while 87-100% of the larvae on Chinese cabbage and common cabbage survived to pupation. Dual choice tests with a Y-tube olfactometer showed that volatiles from B. vulgaris were much more attractive to P. xylostella adults than those from common cabbage. The results demonstrate that B. vulgaris has a great potential as a dead-end trap crop for improving management of P. xylostella. Factors that may influence the feasibility of using B. vulgaris as a trap crop in the field are discussed, and ways to utilize this plant are proposed. PMID- 15541191 TI - Laboratory adaptation of Bactrocera tryoni (Diptera: Tephritidae) decreases mating age and increases protein consumption and number of eggs produced per milligram of protein. AB - A significant reduction in age of mating occurred during the first four generations (G1-G4) of laboratory adaptation of wild Bactrocera tryoni (Froggatt) and this was associated with the earlier attainment of peak egg load although no significant differences were detected in the peak egg load itself. A long term laboratory (LTL) strain had a significantly earlier mating age and higher peak egg load than flies of wild origin or those from the first four laboratory generations. The amount of protein consumed by females in the first week of adult life was significantly higher in the LTL strain than in flies of wild origin or G1-G4 but there were no significant changes (or only slight changes) with laboratory adaptation in the amounts of protein consumed up to the ages of mating and peak egg load. Laboratory adaptation resulted in no significant changes in egg size, egg dry weight, puparial fresh weight and the dry weight of newly emerged females. The large increase in fecundity with laboratory adaptation is associated with a 4- to 5-fold increase in the rate of conversion of dietary protein to eggs (i.e. eggs produced per mg of protein consumed). PMID- 15541192 TI - Mosquitoes of the Anopheles maculipennis group (Diptera: Culicidae) in Romania, with the discovery and formal recognition of a new species based on molecular and morphological evidence. AB - Mosquitoes of the Anopheles maculipennis group were collected in five districts of Romania (Constant,a, Giurgiu, Ilfov, Mehedint,i and Suceava) between March 2000 and June 2003. Two hundred and ninety-seven specimens were identified by molecular methods. Nuclear rDNA ITS2 sequences of 178 specimens were compared with GenBank sequences for nine known Palaearctic species of the group, and 119 specimens were identified using an ITS2 PCR-RFLP assay developed during the study. Five genetically distinct species of the group were identified: A. atroparvus van Thiel, A. maculipennis Meigen, A. melanoon Hackett and A. messeae Falleroni and a previously unrecognized species. The new species, herein formally described and named A. daciae sp. n., was collected in the Black Sea coastal region and plains adjacent to the Danube River in southern Romania. Anopheles daciae is most similar to and sympatric with A. messeae. It is contrasted with A. messeae and characterized on the basis of unique nuclear ITS2 and mitochondrial COI DNA sequences and morphological characters of the eggs. The larval, pupal and adults stages of the two species were also compared, but no reliable characters were found to distinguish them. It seems likely that A. daciae is more widespread in eastern Europe and the Balkan States, and could be responsible for malaria transmission in these regions that is currently attributed to A. messeae. Anopheles melanoon is reported from Romania for the first time. PMID- 15541193 TI - The classification of genus Anopheles (Diptera: Culicidae): a working hypothesis of phylogenetic relationships. AB - The internal classification of genus Anopheles is updated to reflect taxonomic actions published since the classification was last reviewed in 1994. Both formal and informal taxa are included. The classification is intended to aid researchers and students who are interested in analysing species relationships, making group comparisons and testing phylogenetic hypotheses. The genus includes 444 formally named and 40 provisionally designated extant species divided between six subgenera: Anopheles, Cellia, Kerteszia, Lophopodomyia, Nyssorhynchus and Stethomyia. Subgenera Anopheles, Cellia and Nyssorhynchus are subdivided hierarchically into nested informal groups of morphologically similar species that are believed to represent monophyletic lineages based on morphological similarity. Changes to the classification include additional species, eliminated species and changes to the hierarchical organization and composition of supraspecific groups, some as a result of molecular studies. PMID- 15541194 TI - Biological control of house flies Musca domestica and stable flies Stomoxys calcitrans(Diptera: Muscidae) by means of inundative releases of Spalangia cameroni(Hymenoptera: Pteromalidae). AB - The efficacy of the pupal parasitoid Spalangia cameroni Perkins as a biological control agent was tested against house flies Musca domestica Linnaeus and stable flies Stomoxys calcitrans (Linnaeus) in one dairy cattle and two pig installations in Denmark. Weekly releases of S. cameroni from April through to September-October 1999 and 2000 resulted in significant suppressions of house fly populations to below nuisance level, whereas no effect on stable flies was found. Parasitism was significantly higher in the release years compared to the control years, but was below 25% averaged over the fly season for each farm. A statistical model based on a functional relationship between the innate capacity of increase of the two fly species and three explanatory variables (air temperature, fly density and parasitism) provided a fairly good fit to data with the abundances of house flies and stable flies explained mostly by temperature, but intra- and interspecific competition, and parasitism had a significant effect as well. Overall, the model was capable of explaining 14% and 6.6% of the total variation in data for house fly and stable fly, respectively. Spalangia cameroni was the predominant parasitoid to emerge from exposed house fly pupae, but from mid summer onwards Muscidifurax raptor Girault & Sanders (Hymenoptera: Pteromalidae) was also quite common. The study indicated that biological control of house flies can be an efficient alternative to chemical control. PMID- 15541195 TI - Sex ratios of Sitodiplosis mosellana (Diptera: Cecidomyiidae): implications for pest management in wheat (Poaceae). AB - Sex ratios of populations of the wheat midge Sitodiplosis mosellana Gehin, developing on wheat Triticum aestivum L., were determined at reproduction, adult emergence, and dispersal. The patterns of sex ratio through the life cycle of S. mosellana result from: (i) a genetic mechanism that causes all or nearly all of the progeny of individual females to be a single sex, with an overall sex ratio that is slightly biased at 54-57% females; (ii) a differential mortality during diapause that increases the sex ratio to 60-65% females; (iii) mating which occurs near the emergence site followed by female dispersal which causes the post dispersal sex ratio to rise to nearly 100% females; and (iv) oviposition which spreads eggs among different plants and assures that the next generation has a local sex ratio close to the population average. These changes in sex ratio through the life cycle have implications for using crop resistance or pheromones to manage S. mosellana, because mating takes place quickly near emergence sites, and because mated females but not males disperse from emergence sites to oviposition sites. Crop refuges used to protect resistance genes against the evolution of virulence by S. mosellana must be interspersed to prevent assortative mating that would occur in separate blocks of resistant and susceptible plants. Monitoring or mating disruption using a pheromone would be ineffective when wheat is grown in rotation with a non-host crop. PMID- 15541196 TI - Ontogenetic shifts in intraguild predation on thrips by phytoseiid mites: the relevance of body size and diet specialization. AB - In greenhouse agroecosystems, a guild of spider mite predators may consist of the oligophagous predatory mite Phytoseiulus persimilis Athias-Henriot, the polyphagous predatory mite Neoseiulus californicus McGregor (both Acari: Phytoseiidae) and the primarily herbivorous but facultatively predatory western flower thrips Frankliniella occidentalis Pergande (Thysanoptera: Thripidae). Diet specialization and the predator body size relative to prey are crucial factors in predation on F. occidentalis by P. persimilis and N. californicus. Here, it was tested whether the relevance of these factors changes during predator ontogeny. First, the predator (protonymphs and adult females of P. persimilis and N. californicus): prey (F. occidentalis first instars) body size ratios were measured. Second, the aggressiveness of P. persimilis and N. californicus towards F. occidentalis was assessed. Third, survival, development and oviposition of P. persimilis and N. californicus with F. occidentalis prey was determined. The body size ranking was P. persimilis females > N. californicus females > P. persimilis protonymphs > N. californicus protonymphs. Neoseiulus californicus females were the most aggressive predators, followed by highly aggressive N. californicus protonymphs and moderately aggressive P. persimilis protonymphs. Phytoseiulus persimilis females did not attack thrips. Frankliniella occidentalis larvae are an alternative prey for juvenile N. californicus and P. persimilis, enabling them to reach adulthood. Females of N. californicus but not P. persimilis sustained egg production with thrips prey. Within the guild studied here, N. californicus females are the most harmful predators for F. occidentalis larvae, followed by N. californicus and P. persimilis juveniles. Phytoseiulus persimilis females are harmless to F. occidentalis. PMID- 15541197 TI - Infectious mononucleosis and Epstein-Barr virus. AB - Epstein-Barr virus (EBV) is a gamma-herpesvirus that infects over 90% of the human population worldwide. It is usually transmitted between individuals in saliva, and establishes replicative infection within the oropharynx as well as life-long latent infection of B cells. Primary EBV infection generally occurs during early childhood and is asymptomatic. If delayed until adolescence or later, it can be associated with the clinical syndrome of infectious mononucleosis (also known as glandular fever or 'mono'), an illness characterised by fevers, pharyngitis, lymphadenopathy and malaise. EBV infection is also associated with the development of EBV-associated lymphoid or epithelial cell malignancies in a small proportion of individuals. This review focuses on primary EBV infection in individuals suffering from infectious mononucleosis. It discusses the mechanism by which EBV establishes infection within its human host and the primary immune response that it elicits. It describes the spectrum of clinical disease that can accompany primary infection and summarises studies that are leading to the development of a vaccine designed to prevent infectious mononucleosis. PMID- 15541198 TI - Left ventricular mechanoenergetics in small animals. AB - Studies on left ventricular mechanical work and energetics in rat and mouse hearts are reviewed. First, left ventricular linear end-systolic pressure-volume relation (ESPVR) and curved end-diastolic pressure-volume relation (EDPVR) in canine hearts and left ventricular curved ESPVR and curved EDPVR in rat hearts are reviewed. Second, as an index for total mechanical energy per beat in rat hearts as in canine hearts, a systolic pressure-volume area (PVA) is proposed. By the use of our original system for measuring continuous oxygen consumption for rat left ventricular mechanical work, the linear left ventricular myocardial oxygen consumption per beat (VO2)-PVA relation is obtained as in canine hearts. The slope of VO2-PVA relation (oxygen cost of PVA) indicates a ratio of chemomechanical energy transduction. VO2 intercept (PVA-independent VO2) indicates the summation of oxygen consumption for Ca2+ handling in excitation contraction coupling and for basal metabolism. An equivalent maximal elastance (eEmax) is proposed as a new left ventricular contractility index based on PVA at the midrange left ventricular volume. The slope of the linear relation between PVA-independent VO2 and eEmax (oxygen cost of eEmax) indicates changes in oxygen consumption for Ca2+ handling in excitation-contraction coupling per unit changes in left ventricular contractility. The key framework of VO2-PVA-eEmax can give us a better understanding for the biology and mechanisms of physiological and various failing rat heart models in terms of mechanical work and energetics. PMID- 15541199 TI - Characterization of junctional sarcoplasmic reticulum Ca2+ content and release by short-term mechanical restitution in cardiac muscle. AB - To study Ca2+ handling by the junctional sarcoplasmic reticulum (JSR), the time course of short-term mechanical restitution after varying magnitudes of twitch contractions was assessed in rat papillary muscle. Mechanical restitution consisted of a pretwitch latency period followed by a rapid and a subsequent much slower restitution of twitch force. The rate of rapid restitution was independent of the magnitude of the preceding twitch, which suggests that the rate of JSR Ca2+ repletion was dependent on the amount of Ca2+ remaining in the JSR after a twitch contraction. Based on this finding, the functions Gt and Ht, representing the time courses of JSR Ca2+ repletion and release, respectively, were derived graphically from a family of the mechanical restitution curves. Gt increased monotonically with time at a decreasing rate, while Ht increased with time in a sigmoid manner. The mechanical alternans were simulated by using experimental values and mathematically predicted values of Gt and Ht. A substitution of extracellular Na+ with Li+ to inhibit Na+/Ca2+ exchange resulted in an augmentation of Gt by approximately 10%, presumably by increasing the tubular SR Ca2+ uptake. The inhibition of tubular SR Ca2+ uptake by thapsigargin (10 microM) reduced mechanical restitution by approximately 13% of the maximal twitch force, independent of the phase of mechanical restitution; the effect was greater at an earlier time point in the mechanical restitution. These results suggest that early JSR Ca2+ replenishment results mainly from the movement of Ca2+ from the tubular SR. PMID- 15541200 TI - Gastric myoelectrical activity increases after moderate-intensity exercise with no meals under suppressed vagal nerve activity. AB - Postprandial gastric myoelectrical activity recorded by electrogastrogram (EGG) with the subject in a supine position has shown to be enhanced after moderate intensity pedaling exercise in an upright seated position, despite the suppression of vagal nerve activity. However, it is still unknown whether the effect is due to the exercise itself and/or a meal or how the position change has influenced the effects. To address this, we used a position-controllable cycle ergometer to examine the effects of the moderate-intensity exercise on EGG activity and the high-frequency (HF) component of heart rate variability (HRV), an index of vagal nerve activity. To eliminate the effect of position change, we carried out the exercise and the EGG recording in the supine position. The peak amplitude of the EGG was enhanced by prior moderate-intensity exercise with a reduced HF component of HRV, which did not differ for postexercise conditions with or without a meal. The small amount of meal itself, however, enhanced both the peak amplitude of the EGG and the HF component of HRV. The peak frequency of EGG was reduced and the instability coefficient of EGG was increased only after the exercise itself. Taken together, these results suggest that the enhanced amplitude of gastric myoelectrical activity can be induced by moderate-intensity exercise itself, even with suppressed vagal nerve activity, and that the mechanism underlying the exercise effects would differ from that underlying the effect of a meal alone. PMID- 15541201 TI - Hindlimb suspension inhibits air-righting due to altered recruitment of neck and back muscles in rats. AB - Effects of 9-week hindlimb suspension and 8-week recovery on air-righting reaction in response to drop from a supine position were studied in adult rats. The righting time in rats at the end of suspension (approximately 220 ms) was longer than the age-matched controls (approximately 120 ms, p <0.05). The unloading-related change in righting time was accompanied by lowered activities of electromyogram (EMG) and altered recruitment of both neck and back muscles at a specific stage of drop. After 8 weeks of reambulation, righting time recovered toward the control level (approximately 153 ms, p <0.05), but the EMG activity of back muscle was still less than controls. In contrast, the EMG of neck muscle during fall was even increased. The differences in the characteristics of the muscle fibers between two groups were minor. It is suggested that inhibition of recruitment, rather than the changes in the fiber characteristics, of neck and back muscles is one of the major causes of the slow air-righting. PMID- 15541202 TI - Inhibitory effect of pain-eliciting transcutaneous electrical stimulation on vibration-induced finger flexion reflex in the human upper limb. AB - We studied the effects of non-pain transcutaneous electrical stimulation (TES) and pain-eliciting TES on vibration-induced finger flexion reflex (VFR) in 12 healthy volunteers. Tonic finger flexion reflex in the upper limb was induced by the application of vibratory stimulation on the volar side of the middle fingertip in the right hand before and after TES. Non-pain TES or pain-eliciting TES was applied on the skin between the bases of the first and second metacarpals in the right hand dorsal area in a crossover design. Pain-eliciting TES inhibited VFRs significantly (Fisher's PLSD, p <0.01), compared to those of the time control group during and after TES. VFRs were reduced approximately to 63.8% and 78.6% of prestimulation flexion force during and after pain-eliciting TES, respectively. Nonpain TES did not inhibit VFR. These results suggest that pain conducting afferent fibers have inhibitory neuronal connection over the ipsilateral reflex circuits of VFR in the upper limb. PMID- 15541203 TI - A genistein-sensitive Na+/Ca2+ exchange is responsible for the resting [Ca2+]i and most of the Ca2+ plasma membrane fluxes in stimulated rat cerebellar type 1 astrocytes. AB - The differential role of Na+/Ca2+ exchange in the regulation of intracellular ionized calcium ([Ca2+]i) in immunological and pharmacologically identified type 1 astrocytes and Purkinje cells was studied in rat cerebellar culture, using Ca2+ (Fluo-3, Fura-2) and Na+ (SBFI) fluorescence measurements. The mean resting [Ca2+]i was significantly higher (191 +/- 8 nM, n=25) in type 1 astrocytes than in Purkinje cells (92 +/- 2.5 nM, n=35). In contrast to Purkinje cells, in unstimulated cerebellar type 1 astrocytes, forward and reverse Na+/Ca2+ modes operate under resting physiological conditions, being responsible for most of the total Ca2+ transplasma membrane fluxes. Four observations support this hypothesis: (1) under resting conditions of temperature and ionic composition, Na+o removal causes a remarkable increase in [Ca2+]i, being inhibited by 2',4' dichlorobenzamil (DCB), and 2-[2-[4-(nitrobenzilloxiphenyl ethyl] isothiourea metanesulfonate (KB-R7943); (2) Ca2+o removal in the presence of Na+o causes an important drop in [Ca2+]i, which is absent in Li+o or NMG+o (N-methyl-D glucamine) containing medium; (3) the reverse mode exchange inhibitor KB-R7943 mimics the removal of Ca2+o only in the presence of Na+o; and (4) under loaded [Na+]i conditions (ouabain or the activation of taurine-Na+-cotransport), reverse mode exchange increases in both astrocytes and Purkinje cells. In type 1 astrocytes stimulated with endothelin-3 (ET-3), the recovery of the Ca2+i signal occurs largely through the Na+/Ca2+ exchanger. Genistein, a tyrosine kinase inhibitor, completely and reversibly blocks all exchange activity, but not its inactive analogue daidzein, thus suggesting that the Na+/Ca2+ exchanger of cerebellar type 1 astrocytes may be modulated by phosphorylation. Our main conclusion is that in rat cerebellar type 1 astrocytes under resting physiological conditions, most of the total transplasma membrane Ca2+ fluxes take place through the Na+/Ca2+ exchanger, thus accounting for the resting [Ca(2+)]i. PMID- 15541204 TI - Heat-stress enhances proliferative potential in rat soleus muscle. AB - The effects of heat-stress on proliferative potential in vivo were studied in rat skeletal muscle. Male Wistar rats (7-weeks-old) were divided into two groups: control (n=24) and heat-stressed (n=24). Rats in the experimental group were exposed to environmental heat-stress (41 degrees C for 60 min) in a heat chamber without anesthesia. The soleus muscles were dissected 1, 7, and 14 days after the heat exposure. The wet and dry weights of soleus muscle relative to body weight in the heat-stressed group were significantly higher than controls 7 days after the exposure (10.1% and 17.5%, respectively, p <0.05). The distribution of 5 bromo-2'-deoxyuridine and proliferating cell nuclear antigen-positive nuclei, that are the indicators for the cell proliferation, were increased by 2.2 and 5.1 times, respectively 1 day after heating (p <0.05). The expressions of heat shock protein 72 (58.0%) and phosphorylated p70S6 kinase (52.3%) were increased 1 day following heat exposure (p <0.05). These results suggest that heat-stress could promote the cell proliferation and induce muscular hypertrophy. PMID- 15541205 TI - Autonomic heart rate regulation during mild dynamic exercise in humans: insights from respiratory sinus arrhythmia. AB - To better understand the neural mechanism of heart rate (HR) regulation during dynamic exercise, the responses of HR and the magnitude of respiratory R-R interval variation were examined during exercise and recovery at mild intensities in humans. Eight subjects performed 3-min constant load cycle exercises in a semi supine position at work rates of 25, 50, and 100 W. The respiratory interval was fixed at 4 s. Peak-to-valley variation in R-R interval caused by respiration was measured breath-by-breath and standardized for tidal volume (DeltaRRst, a noninvasive index of the degree of parasympathetic cardiac control). At all work rates the HR increased significantly from 2.5 s after the beginning of exercise (p <0.05) and decreased temporarily and slightly at around 15 s, and the DeltaRRst varied almost inversely. The HR and the DeltaRRst until 12.5 s after the beginning of exercise changed independently of work rate (ANOVA, p=0.27 and p=0.08). The HR-DeltaRRst relationship at the initial phase of exercise (for 12.5 s) was almost the same at all work rates. These results suggest that the initial HR response to exercise is strongly parasympathetically regulated independently of work rate. The HR recovered slower than the DeltaRRst at 50 and 100 W. On the HR-DeltaRRst relationship, the HR during recovery was significantly higher than during exercise at 1/3, 1/2, and 2/3 levels of pre-exercise DeltaRRst at 50 and 100 W and at the 1/3 level at 25 W (p < 0.05). At 25 W, the difference in HR at the 1/3 level was 5.5 beats.min(-1), and the HR increase to exercise was 21.2 beats.min(-1). We suggest that a HR regulatory system responds slower than a cardiac parasympathetic system to exercise, a cardiac sympathetic system, is activated even during mild exercise in humans. PMID- 15541206 TI - Heat stress facilitates the recovery of atrophied soleus muscle in rat. AB - Effects of heat stress on the recovery of atrophied soleus muscle were studied in rats. Ten-week-old male Wistar rats were randomly divided into cage control (CC) and 5-day hindlimb suspension group (HS). The half of the rats in group HS was exposed to heat stress (41 degrees C for 60 min) in an incubator immediately after the hindlimb suspension (HS-H) and the other group of rats was not heat stressed (HS-C) prior to 10 days of ambulation recovery. One group of cage control rats (CH) was also exposed to heat similarly. The soleus muscles were dissected at four time points, i.e., immediately after the suspension (or heat stress), and 3, 5, and 10 days after the recovery (n=8 per group at each time point). The absolute wet weight and water and protein content of whole soleus muscle in group HS-C were approximately 36, 27, and 8 mg less than CC (p <0.05). Thus, the percentage contribution of water and protein loss to the decrease in muscle weight was 75 and 22%, respectively. Although water content, as well as muscle weight, was elevated within 3 days, the increase of protein was delayed. Heat exposure prior to recovery accelerated the increase in protein content even in the control group. These phenomena were closely associated with 72-kD heat shock protein (HSP72) content. It is suggested that heat stress applied at the end of hindlimb unloading facilitated the recovery of atrophied soleus muscle of rat, through possibly HSP72-related events of protein metabolism. The data also indicated that the combination of heat and mechanical stress evoked larger and long lasting HSP72 response than does heat or mechanical stress alone. PMID- 15541207 TI - Determination in vivo of newly synthesized gene expression in hamsters during phases of the hibernation cycle. AB - This study measured in vivo synthesis of total RNA and protein from cortex, cerebellum and midbrain/brainstem and 6 major organs from Syrian hamsters (Mesocricetus auratus) during (a) 33 h of torpor (body temperature 5-6 degrees C); (b) 90 min of the early arousal; (c) 90 min of the middle arousal; (d) 90 min in cold adapted cenothermic (CEN) hamsters of the same circannual period. Appropriate physiological parameters were used to confirm the phase of the hibernation cycle during infusion and incorporation of [3H]-uridine and [14C] leucine. In torpor, RNA synthesis was 5-25% of CEN levels depending upon tissue. In brain and heart mRNA was not preferentially synthesized. Protein was synthesized at low, tissue specific levels during torpor. Initiation of arousal and the warming of anterior organs via non-shivering thermogenesis during the early arousal occurred without measurable synthesis of RNA or proteins. Tissue specific levels of RNA and protein synthesis occurred later after shivering thermogenesis had been recruited and was strongly influenced by thermal gradients in the body. In the middle arousal phase, protein synthesis is most active in the brain despite modest synthesis of RNA and mRNA. The majority of molecular processing required for the induction and maintenance of torpor and the arousal from torpor up until the onset of shivering thermogenesis occurs during the cenothermic period before the hamster initiates the hibernation cycle. PMID- 15541208 TI - The urge to define urgency: a review of three approaches. PMID- 15541209 TI - Effect of trospium chloride on somnolence and sleepiness in patients with overactive bladder. AB - An objective of a large US phase-3 study was to determine the effects of trospium chloride 20-mg tablets versus placebo, administered twice daily, on central nervous system (CNS) adverse events (eg, somnolence) and on daytime sleepiness using the validated Stanford Sleepiness Scale. The impact of age groups also was assessed. Trospium chloride did not increase daytime sleepiness or appear to produce CNS adverse effects, as measured by the Stanford Sleepiness Scale and the incidence of CNS adverse events. Furthermore, there was no difference in these findings across age groups. PMID- 15541210 TI - Mixed incontinence: stressing about urge. AB - Urgency and urinary urge incontinence are troublesome and complex symptoms that can complicate the diagnosis and treatment of stress urinary incontinence. Despite this fact, first-line intervention frequently is directed toward the incompetent bladder neck using surgical approaches. Persistent urgency or detrusor instability following anti-incontinence surgery reduces patient satisfaction and overall continence in most series. This article discusses current theories explaining the etiology of mixed incontinence. It also reviews the results of anti-incontinence surgery for mixed incontinence and discusses management strategies. Possible predictors of outcome after sling surgery in this complex group of patients also are presented. PMID- 15541211 TI - A case for botulinum toxin-A in idiopathic bladder overactivity. AB - Botulinum toxin (BTX) has been shown to be an effective agent in suppressing detrusor overactivity due to neurogenic causes. Similar to results obtained with traditional agents to treat bladder overactivity (ie, antimuscarinic medications), the use of BTX has been extended to patients with idiopathic detrusor overactivity. This article briefly reviews the use of BTX to treat disorders of detrusor overactivity and, based on early clinical and laboratory results, establishes the case for its use as a therapeutic modality to treat idiopathic detrusor overactivity. PMID- 15541212 TI - Chronic prostatitis and sensory urgency: whose pain is it? AB - Difficulties encountered in diagnosing and effectively treating chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is frustrating for clinicians and patients. Scientific evidence cannot establish an exact relationship between the prostate and the symptoms of CP/CPPS, and the prostate continues to be the diagnosis of convenience in this complex syndrome in men. However, if the pain is not the prostate's, whose pain is it? A heterogeneous group of insults can result in a common neurogenic pain response, resulting in recurring pain and voiding or sexual dysfunction. To add to this dilemma, certain life-threatening diagnoses, such as carcinoma-in-situ, is in the differential diagnosis and must be excluded. Urodynamics may be useful in evaluating and treating patients whose voiding symptoms predominate. However, many patients with CP/CPPS will not have measurable abnormalities by conventional methods and likely suffer from a functional somatic syndrome that is best treated with a multimodality approach. PMID- 15541213 TI - Confusion about measuring central nervous system effects. AB - Anticholinergic therapy together with behavioral treatment are the main stays of treatment for the overactive bladder. Successful therapy and patient compliance depend very much on side effects. In the past, little attention has been paid to anticholinergic side effects in the central nervous system (CNS), which can be critical, especially for elderly patients. Incidence and intensity of CNS effects depend on the pharmacokinetic and pharmacodynamic properties that are decisive whether anticholinergics pass the blood-brain barrier as a result of passive and active transport mechanisms. To measure potential CNS side effects of anticholinergic drugs, rapid eye movement sleep analysis, quantitative topographic electroencephalogram studies, and psychometric tests were performed. Structural changes in brain morphology resulting from anticholinergics also were analyzed in a post-mortem study. PMID- 15541215 TI - Phosphodiesterase-5 inhibitors: clinical market and basic science comparative studies. AB - The purpose of this review is to examine the biologic, pharmacologic, and clinical differences between the three currently approved phosphodiesterase-5 inhibitors to help the clinician make an educated choice about which medication may be best for any individual patient. PMID- 15541216 TI - Penile prosthesis coating and the reduction of postoperative infection. AB - Despite the recent development of effective oral agents for the treatment of erectile dysfunction, penile prosthesis implantation remains an effective and acceptable treatment for the significant number of men who fail to respond to nonsurgical therapy. The most serious complication that can affect the use of most prosthetic devices is infection. In penile prostheses, this can be devastating and frequently results in removal of the device despite aggressive antibiotic therapy. In recent years, new strategies have been developed in an attempt to minimize this risk. This review focuses on one such method, namely the use of an antibiotic coating on the device. It reviews recent published data regarding the effectiveness of such devices at decreasing infection rates. PMID- 15541217 TI - Erectile dysfunction secondary to nerve-sparing radical retropubic prostatectomy: comparative phosphodiesterase-5 inhibitor efficacy for therapy and novel prevention strategies. AB - Postprostatectomy erectile dysfunction appears to be initiated by neuropraxia and perpetuated by cavernosal smooth muscle apoptosis. Phosphodiesterase-5 (PDE-5) inhibitor therapy is the current cornerstone of erectile dysfunction (ED) therapy in this population. Although no head-to-head trials have been performed with sildenafil, vardenafil, and tadalafil in this population, there are numerous studies in the general ED population. The results of these studies demonstrate that neither of the new PDE-5 inhibitors met statistical noninferiority to sildenafil. Sildenafil has been studied in a novel primary prevention modality using nightly administration after a bilateral nerve-sparing prostatectomy. In this novel approach, it effected a sevenfold improvement in return of spontaneous, normal erectile function 2 months after drug discontinuation. This effect appears to be mediated by properties unique to sildenafil that include improved endothelial function and neuronal regeneration and neuroprotection. In primary prevention, unlike ED therapy, one has only "one shot" by definition. Therefore, it is even more critical to apply evidence-based medicine. PMID- 15541218 TI - Testosterone treatment for the aging man: the controversy. AB - The issues surrounding androgen replacement therapy in men with symptomatic late onset hypogonadism has been marred in controversy even before the identification and synthesis of testosterone. The controversy has attained renewed importance because of the ever increasing aging population. Physicians attuned with the various diagnostic approaches, familiar with the advantages and drawbacks of testosterone preparations, and aware of the potential safety issues are well positioned to deal with the hypogonadal man at or beyond middle age. The urologist is in a unique position to assist patients and colleagues in other specialties, particularly in relation to prostate safety. This update focuses on the most prominent areas of controversy. PMID- 15541219 TI - New discoveries in the basic science understanding of Peyronie's disease. AB - Peyronie's disease is an acquired benign condition representing localized fibrosis of the penis. The disease is caused by microvascular trauma to the penis, with subsequent fibrin deposition resulting in aberrant wound healing and possibly other etiologies. Histopathologic studies of the Peyronie's plaque have demonstrated the role of transforming growth factor-beta 1 in the pathologic development of penile plaque. Animal models have been developed to test pathophysiology and therapeutics. The role of inducible nitric oxide synthase and nitric oxide has been established to limit and counteract fibrosis. Differential gene expression studies have identified candidate up-regulated or down-regulated genes that are involved in the pathophysiology of Peyronie's disease. PMID- 15541220 TI - Safety and immunogenicity of a new chromatographically purified rabies vaccine in comparison to the human diploid cell vaccine. AB - BACKGROUND: Although human diploid cell vaccine (HDCV) has been available for over two decades and has a proven record of efficacy, it is very expensive to produce and can only be made in small quantities. METHODS: In this trial, we compared the safety and immunogenicity of a new, chromatographically purified rabies vaccine (CPRV) with those of HDCV. One hundred and thirty-five healthy veterinary students were randomized in a 2:1 ratio between CPRV and HDCV respectively. Each student subsequently received an intramuscular injection of 0.5 mL of CPRV or 1mL of HDCV on days 0, 7, and 28, according to the standard preexposure regimen. Local safety data were collected for 7 days following each dose and systemic safety data for 42 days following the first dose. Vaccine administration and safety evaluation were performed by different site personnel. Sera for immunogenicity analysis were collected on days 0 (prevaccination), 28 and 42. RESULTS: All subjects achieved an antirabies antibody titer greater than or equal to the World Health Organization (WHO) accepted threshold level of seroconversion of 0.5 IU/mL after only two of three doses of vaccine in both groups. The geometric mean titers (IU/mL) in the CPRV and HDCV groups respectively were 6.54 (range 0.50 to 64.80) and 10.22 (range 0.70 to 51.40) on day 28, and 40.51 (range 5.40 to 278.00) and 37.71 (range 5.40 to 278.00) on day 42. The percentage of subjects experiencing local reactions within 3 days after any dose ranged from 65.2% to 80.9% in the CPRV group and from 77.3% to 84.4% in the HDCV group. The local reaction reported by the greatest percentage of subjects after each dose was pain/tenderness at the injection site, and most reactions were mild. Most of the reported local reactions resolved within 0 to 3 days postvaccination. Systemic reactions decreased from 76.4% after dose 1 to 36.0% after dose 3 in the CPRV group, and similarly from 55.6% to 31.8% in the HDCV group. For all postdose periods, the systemic reaction reported by the highest percentage of subjects was myalgia. No subjects experienced an immediate local or systemic reaction. CONCLUSIONS: In healthy adults, vaccination with CPRV using a preexposure schedule resulted in a safety and immunogenicity profile similar to that of HDCV. PMID- 15541221 TI - Immunogenicity of booster vaccination with a virosomal hepatitis A vaccine after primary immunization with an aluminum-adsorbed hepatitis A vaccine. AB - BACKGROUND: Increasing numbers of individuals are traveling to areas of high hepatitis A endemicity and require immunization against the hepatitis A virus (HAV). The option of using a virosomal, aluminum-free, HAV vaccine (Epaxal) for booster immunization following primary vaccination with an aluminum-adsorbed vaccine has been assessed. METHODS: In total, 142 healthy subjects, 79 men and 63 women, aged 12 to 72 years, were injected intramuscularly with a booster dose of Epaxal (0.5 mL containing < or =500 RIA units of HAV antigen) 6 to 24 months after primary vaccination with Havrix (0.5 or 1.0 mL containing 720 or 1440 ELISA units of HAV antigen, respectively, adsorbed onto aluminum hydroxide). Anti-HAV antibody titers were measured on days 0 and 28 by an enzyme immunoassay. Adverse events were recorded for 1 month postinjection. RESULTS: Overall, 98/118 subjects (83%) with no serologic evidence of past HAV infection were still seroprotected at enrolment (anti-HAV antibody titer < or = 20 mIU/mL). The seroprotection rate was 87% in those primed with Havrix 1440 6 to 12 months earlier (n=93) and 60% in those primed < or =12 months before enrolment (n=20, mean 16 months). The geometric mean anti-HAV antibody titer increased from 65 mIU/mL at day 0 to 1,722 mIU/mL at day 28 after a single booster dose with Epaxal in evaluable subjects who were primarily vaccinated with either a single dose of Havrix 1440 (n=111) or two separate doses of Havrix 720 (n=4). All subjects were seroprotected at day 28, and 98% showed at least a four-fold increase in anti-HAV antibody titer. Epaxal was well tolerated and no serious adverse events were reported. At day 28, the tolerability of the vaccination was judged as either "very good" or "good" by 96% of vaccinees and by all investigators. CONCLUSION: Epaxal can be successfully used to boost immunization following primary vaccination with an aluminum adsorbed vaccine, and is well tolerated. PMID- 15541222 TI - Medical supplies for travelers to developing countries. AB - BACKGROUND: There has been little research to date on the use of medicines and first aid supplies by travelers. In some developing countries such products may be difficult to obtain, and there is the danger that substandard medicines may be purchased. As space for medical supplies in the luggage of many individuals, particularly backpackers, may be restricted, it is important to identify correctly those items most likely to be needed. OBJECTIVES: The aim of this study was to survey a cohort of travelers from the UK visiting a variety of destinations in developing countries, regarding the medical supplies taken and used during their trip. METHOD: Travelers visiting a specialist travel pharmacy in London, UK were recruited consecutively into the study. Only those planning to visit destinations in South America, Asia, Africa or the Middle East for < or = 2 weeks and returning to the UK were included. Participants were handed a questionnaire to be posted back when they returned to the UK, asking them to note those items that they included in the kit, those actually used, and any others obtained while they were away. All subjects had consulted the pharmacist concerning the medical kit appropriate for their trip. RESULTS: Two hundred and ninety-nine travelers volunteered to take part, of whom 127 returned the postal questionnaire. Analgesics and medication for the treatment of diarrhea were most likely to be used, but many types of wound dressing were unlikely to be required. Twenty individuals (16%) required antibiotics, with eight people purchasing them while they were away. Thirty-two (31%) individuals did not use insect repellents despite traveling to potentially malaria-endemic countries. Only seven subjects purchased any other items while they were away. CONCLUSION: The items most likely to be required by travelers to developing countries are analgesics, treatments for diarrhea, antiseptics and sticking plasters. The provision of antibiotics to certain travelers is probably justified. PMID- 15541223 TI - Imported Plasmodium vivax malaria: demographic and clinical features in nonimmune travelers. AB - BACKGROUND: Imported malaria is an important problem in nonendemic countries due to increasing travel to and immigration from malaria-endemic countries. Plasmodium vivax malaria is relatively common in travelers but there are few published data regarding the outcome of P. vivax malaria in this group. METHODS: We analyzed 209 cases of P. vivax malaria that were reported to the GeoSentinel network and the VIDS database, Royal Melbourne Hospital. Analyses were performed on data including demographics, pretravel encounter, antimalarial prophylaxis, exposure history, type of travel, countries of recent and past travel, clinical presentation, treatment, outcome and final diagnoses. RESULTS: The majority of patients were travelers (61%), followed by expatriates (13%) and recent immigrants or foreign visitors (12%). Recent travel to Oceania, sub-Saharan Africa, and South and Central America was significantly more likely to be associated with P. vivax malaria than travel to all other regions. The clinical presentation of P. vivax malaria acquired in the Pacific region is indistinguishable from infection with P. falciparum. The use of chloroquine prophylaxis did not prolong the incubation period. Relapse of infection was not infrequent, and the only significant predictor of relapse was travel to Papua New Guinea (PNG), regardless of primaquine dose. Travelers returning from PNG were eight times more likely to relapse after primaquine treatment compared to travelers with P. vivax malaria acquired elsewhere. CONCLUSIONS: We have presented details of the epidemiology, clinical presentation and management of infection with P. vivax malaria in travelers. P. vivax malaria is an important cause of morbidity in travelers, and relapse following primaquine treatment is especially problematic with P. vivax malaria acquired in PNG. PMID- 15541224 TI - Atovaquone-proguanil (malarone): an effective treatment for uncomplicated Plasmodium falciparum malaria in travelers from Denmark. AB - BACKGROUND: Previous experience with unacceptable adverse effects with mefloquine as treatment for uncomplicated Plasmodium falciparum malaria prompted an evaluation of the effectiveness and side effects of atovaquone-proguanil (Malarone) in a hospital setting. METHODS: Atovaquone-proguanil was given as standard treatment (1,000/400 mgq.d. for 3 days) to 50 adults who had traveled in Africa and returned with uncomplicated Plasmodium falciparum malaria. Half of the treated patients were African and had lived outside Africa for varying periods of time; the other half were Danish-born persons without any previous immunity towards malaria. RESULTS: All patients treated with Malarone were cured without complications. The mean fever clearance times differed among the groups and according to various degrees of prior exposure to malaria and ranged from 1.3 to 2.2 days. Adverse effects during treatment were mild, and were likely to be due to the malaria itself. Fourteen people who had acquired falciparum malaria in spite of taking proguanil-chloroquine prophylaxis were also cured uneventfully without recrudescence. CONCLUSIONS: Malarone appears to be an effective, safe and acceptable oral treatment for uncomplicated malaria. PMID- 15541225 TI - African tick-bite fever: four cases among Swiss travelers returning from South Africa. AB - BACKGROUND: African tick-bite fever (ATBF) is a recently described disease belonging to the spotted fever group. It is caused by Rickettsia africae, and cases are mainly diagnosed in travelers returning from sub-Saharan Africa. METHODS: We report four cases of ATBF among Swiss travelers returning from a 1 month trip in rural South Africa. Diagnosis was made on the basis of clinical, epidemiologic and serologic findings that we describe in detail. Serology was performed using microimmunofluorescence (MIF) assay 2 weeks, 6 weeks and 14 months after the commencement of symptoms. RESULTS: All patients developed the typical eschar and a rash; two had a local lymphadenopathy and one a lymphangitic reaction. Two patients developed transient neuropsychiatric symptoms such as headache, irritability and depressed mood. All four patients had rises in both IgM and IgG classes of anti-R. africae antibodies. After 1 year, only two patients still had measurable circulating antibodies. Cross-reactions with R. conorii were noted. Three patients were cured after a short course of doxycycline; one required 15 days of treatment. CONCLUSIONS: ATBF is a benign disease increasingly being diagnosed in travelers. After ruling out malaria, ATBF diagnosis relies upon a detailed travel history and the classical findings of influenza-like symptoms, fever, one or more necrotic eschars, and rash. Serologic tests usually help to confirm the diagnosis. Neuropsychiatric symptoms specifically associated with ATBF are reported here for the first time. PMID- 15541226 TI - Epidemiology of travelers' diarrhea: details of a global survey. AB - BACKGROUND: Recent epidemiologic data on travelers' diarrhea (TD) are essential for the evaluation of conventional and future prophylactic and therapeutic measures. METHODS: To determine the epidemiology, including risk factors, impact and quality-of-life evaluation of TD, a cross-sectional survey was conducted over 12 months at the airports of Mombasa (Kenya), Goa (India), Montego Bay (Jamaica) and Fortaleza (Brazil) by distributing questionnaires to visitors just prior to their flying home. The study period was March 1996 to July 1998. RESULTS: Overall, 73,630 short-term visitors completed a questionnaire. The total diarrhea attack rate varied between a high of 54.6% in Mombasa and a low of 13.6% in Fortaleza, but only between 31.5% and 5.4% of all travelers had classic TD. The 14-day incidence rates varied between 19.5% and 65.7%. Few travelers meticulously avoided potentially dangerous food items, although in India and Kenya most travelers avoided those considered most dangerous. Risk factors were stays exceeding 1 week, age between 15 and 30 years, and residence in the UK. The impact, measured as incapacity or quality-of-life scores, was very considerable. CONCLUSIONS: TD continues to affect vacationers and business travelers as frequently as it did some 20 years ago. Compliance with recommendations to reduce exposure to pathogens by avoiding dangerous food items is poor among travelers from all countries. Implementation of food safety education programs may be difficult to achieve. PMID- 15541227 TI - Bismuth revisited: an effective way to prevent travelers' diarrhea. PMID- 15541228 TI - Air travel and cardiovascular disease. PMID- 15541229 TI - Ectopic schistosomiasis in a returned traveler. PMID- 15541230 TI - Seroprevalence of malaria infections in Korean troops on a peacekeeping mission in East Timor from 2001 to 2002. PMID- 15541231 TI - Protracted fever after a journey to India and Nepal: a case of persistent Salmonella paratyphi infection. PMID- 15541232 TI - Accelerated vaccination schedules provide protection against hepatitis A and B in last-minute travelers. PMID- 15541233 TI - Transparency in publishing. PMID- 15541235 TI - Usefulness of real-time myocardial perfusion imaging in the evaluation of patients with first time chest pain. AB - In patients who have acute coronary syndrome (ACS), rapid and accurate risk stratification is crucial. Real-time myocardial contrast echocardiography (MCE) extends the evaluation of wall motion abnormalities by assessing myocardial perfusion. We investigated whether MCE could contribute to clinical and biochemical markers in identifying patients who have ACS when presenting to the emergency department. Consecutive patients (n = 100) who presented with first occurrence of chest pain underwent MCE to evaluate myocardial perfusion. Contrast images were also analyzed quantitatively off-line by measuring peak signal intensity (A) and slope of signal intensity increase (beta) in 16 myocardial segments. Thirty-seven of 100 patients had ACS. MCE showed perfusion defects in 9 of 12 patients (75%) who had unstable angina and had high-grade stenotic lesions on an angiogram that were missed by assessment of troponin T. MCE identified all 6 patients who had non-ST-elevation myocardial infarction and no initial increase in troponin T and 17 of 19 patients who had non-ST-elevation myocardial infarction and an initial increase in troponin T. In 2 patients who had chest pain and increased troponin T, MCE excluded ACS by identifying perimyocarditis as the underlying cause. Multivariate logistic regression analysis showed that MCE was the strongest predictor of ACS, thus adding significant diagnostic value to conventional tests. Initial perfusion defect size correlated strongly with increased troponin T at 96 hours (r = 0.73, p <0.001) and with ejection fraction at 4 weeks of follow-up (r = -0.79, p <0.001). Thus, our data suggest that MCE can accurately identify patients who have ACS. PMID- 15541236 TI - Comparison of stress-induced myocardial ischemia in patients with and without coronary arterial collaterals. AB - Coronary artery collaterals may have a protective role against myocardial ischemia at rest. However, their role during stress is controversial and poorly defined. We evaluated 212 consecutive patients (57 +/- 16 years; 70% men) who underwent coronary angiography and stress echocardiography within a 3-month period. This cohort of patients (all had significant epicardial coronary artery disease) was divided into 3 groups based on the presence and type of collaterals: group A, no collaterals; group B, collaterals supplied by vessels without flow limiting stenosis; and group C, collaterals supplied by vessels with flow limiting stenosis. In all patients, angiographic jeopardy score, ejection fraction, and regional and cumulative wall motion score indexes (WMSIs) at rest and during stress were evaluated. Angiographic jeopardy scores were 2.9, 3.5, and 7.3 for groups A, B, and C, respectively. Baseline ejection fraction was similar between groups A and B (48% vs 46%, p = NS) but lower in group C (31%, p <0.01). During stress echocardiography, all groups demonstrated a significant increase in WMSI from baseline (WMSI at rest vs that during stress 1.72 +/- 0.06 vs 1.79 +/- 0.04 for group A, 1.97 +/- 0.06 vs 2.09 +/- 0.03 for group B, 2.35 +/- 0.11 vs 2.41 +/- 0.07 for group C; p <0.01 for all groups). In addition, the number of ischemic segments increased significantly in all groups (2.96 +/- 0.07 in group A vs 4.52 +/- 0.11 in group B vs 5.61 +/- 0.13 in group C, p <0.01). Coronary artery collaterals do not offer protection against stress-induced myocardial ischemia. Moreover, the presence of jeopardized angiographic coronary collaterals (group C) is associated with a higher angiographic jeopardy score, higher baseline WMSI, and decreased ejection fraction. PMID- 15541237 TI - Angiographic and three-dimensional intravascular ultrasound analysis of combined intracoronary beta radiation and self-expanding stent implantation in human coronary arteries. AB - This study tested the combination of vascular brachytherapy (VBT) and self expanding Wallstent implantation in coronary lesions of patients at high risk for restenosis as assessed angiographically by quantitative coronary analysis and by 3-dimensional intravascular ultrasound analysis. Twenty-nine "de novo" lesions were managed with a self-expanding stent alone (n = 19) or with a self-expanding stent after beta-VBT (n = 10) in 27 patients who had been identified by high levels of plasma angiotensin-converting enzyme as being prone to myointimal growth after stent implantation. At 6 months, the increase in stent strut diameter was similar in the 2 groups by quantitative coronary analysis and 3 dimensional intravascular ultrasound (Delta mean stent strut diameter -0.33 +/- 0.3 vs -0.40 +/- 0.3 mm, p = 0.5; Delta stent area -11.8 +/- 6.1 vs -12.0 +/- 6.1 mm(2), p = 0.9; Delta stent volume -96.9 +/- 112 vs -83.5 +/- 73 mm(3), p = 0.7; for groups treated with VBT and self-expanding stents and only self-expanding stents, respectively). In-stent neointimal proliferation was decreased in the group treated with VBT and self-expanding stents (minimal luminal diameter 2.5 +/ 0.8 vs 1.88 +/- 0.8 mm, p = 0.04) by quantitative coronary analysis (minimal luminal area 6.7 +/- 2.5 vs 4.1 +/- 1.9 mm(2), p = 0.01), by intravascular ultrasound, and proliferation volume (84.6 +/- 66.4 vs 159.2 +/- 103.5 mm(3), p = 0.05) by 3-dimensional intravascular ultrasound. Positive vessel and luminal remodelings were observed in 50% of the group treated with VBT and self-expanding stents and in 11% of the group treated only with self-expanding stents (p = 0.02). The combined use of VBT and self-expanding stents is a novel approach that enlarges vascular lumen by preventing vessel constriction and neointimal proliferation. The feasibility and good results of this experimental approach suggest that the simultaneous use of these 2 technologies may be an interesting alternative for difficult vascular districts with high restenosis rates, such as peripheral circulation in the lower limbs. PMID- 15541238 TI - A Consumer Use Study of Over-The-Counter lovastatin (CUSTOM). AB - The Consumer Use Study of OTC Mevacor evaluated the ability of subjects to self manage high levels of low-density lipoprotein (LDL) cholesterol by using a multifaceted cholesterol self-management program (the Mevacor Over-the-Counter Self-Management System; MOTC-SMS). This 26-week all-comers multicenter observational study was conducted in naturalistic storefront settings that used the fully functional MOTC-SMS to guide subjects' behavior. Of 3,316 subjects who evaluated the product (evaluators), 1,061 took >or=1 20-mg tablet of Mevacor OTC (users). Eighty-four percent of evaluators made appropriate initial use decisions. Most users demonstrated acceptable ongoing use behavior regarding treatment to goal, compliance/persistence, changes in health status, dietary patterns, and exercise habits. Throughout the study, 23 users (2%) demonstrated behavior that created the potential for suboptimal safety. After 26 weeks, median levels of LDL cholesterol were reduced by 25% among users who fasted. Of the 878 users who completed the study lipid test, 548 (62%) achieved the LDL cholesterol target goal (<130 mg/dl). Physician interactions were common. Mevacor OTC was well tolerated, with no observable adverse experiences from drug interactions or reports of myopathy. This actual use study demonstrates that the MOTC-SMS can effectively guide consumers to interact with health care professionals and to make appropriate initial and ongoing use decisions to manage their elevated levels of LDL cholesterol, with minimal potential or actual safety risk. PMID- 15541239 TI - Prevalence of coronary heart disease and carotid arterial thickening in patients with the metabolic syndrome (The ARIC Study). AB - We determined the prevalence of the metabolic syndrome (MS) with the criteria recommended by the National Cholesterol and Education Program, Adult Treatment Panel III report and estimated the magnitude of cross-sectional associations between the MS, coronary heart disease (CHD), and atherosclerosis in 14,502 black and white middle-age patients in the Atherosclerosis Risk in Communities Study. CHD was ascertained by standardized procedures and subclinical atherosclerosis was determined by measuring carotid intimal medial wall thickness using B-mode ultrasonography. The prevalence of MS was 30%, with substantial variation across race and gender subgroups. Among women but not among men, MS was significantly associated with increasing low-density lipoprotein cholesterol. CHD prevalence was 7.4% among those with the MS compared with 3.6% in comparison subjects (p <0.0001). After adjustment for established risk factors, subjects who had MS were 2 times more likely to have prevalent CHD than were those who did not have the syndrome. Among individuals free of CHD and stroke, after adjustment for age, gender, and race/center, the average intimal-medial wall thickness of carotid arteries was greater among those with versus those without MS (747 vs 704 mum, p <0.0001). Thus, MS was significantly associated with the presence of CHD and carotid intimal medial wall thickness. Identification of patients who have MS may provide opportunities to initiate CHD prevention strategies. PMID- 15541240 TI - Frequency of elevation of C-reactive protein in atrial fibrillation. AB - Inflammation has been implicated in the pathogenesis of cardiovascular diseases. C-reactive protein (CRP), a marker of systemic inflammation, predicts the risk of coronary events and stroke. Atrial fibrillation (AF) is associated with atrial structural changes that may have an inflammatory basis. We tested the hypothesis that CRP is a risk factor for AF. Subjects were those included in the database registry of the Intermountain Heart Collaborative Study from 1994 to 2001. Patients who had >or=1 electrocardiogram that demonstrated AF formed the disease group (n = 347), and those who had neither electrocardiographic nor clinical evidence for AF comprised the control group (n = 2,449). Logistic regression assessed the quartile (Q) of CRP and 13 other clinical and angiographic predictors of AF. Average age was 63 +/- 12 years, 33% were women, and 61% had advanced coronary artery disease. Patients who had AF were older (by 7 years) and more frequently had a history of heart failure than did controls (41% vs 9%). CRP was higher in patients who had AF than in controls (p <0.001). Q-CRP was a univariable predictor of AF (odds ratio 1.39/Q, 95% confidence interval 1.25 to 1.55, p <0.001). Adjusting for age and heart failure decreased the predictive value of Q-CRP to 1.20/Q (95% confidence 1.07 to 1.34, p = 0.002), whereas further adjustment for 11 other variables had little additional effect (odds ratio 1.19/Q, 95% confidence interval 1.06 to 1.33, p = 0.003). Thus, high levels of CRP independently predicted an increased risk of AF among a large, prospectively studied patient cohort that was assessed angiographically. Increased CRP is a new risk marker for AF propensity, and testing therapies that target inflammation should be considered. PMID- 15541241 TI - Dietary recommendations in the prevention and treatment of coronary heart disease: do we have the ideal diet yet? AB - To respond to the question of the best "heart-healthy" diet, we reviewed the effects of common diets on lipids, their efficacy, advantages, and limitations. The high-protein, low-carbohydrate diet is effective for weight loss over the short term, but its long-term benefits remain unproved. The very low-fat diet decreases levels of total and low-density lipoprotein cholesterol and, with lifestyle modifications, may slow progression of coronary atherosclerosis. The high-protein and very low-fat diets are difficult to follow over the long term. The American Heart Association diet, which is rich in fruits, vegetables, and nuts, decreases blood pressure and may be acceptable to most patients. However, it is rich in carbohydrates and may not be suitable for patients who are obese and/or have high levels of triglycerides. In such patients, diet based on foods with a low glycemic index may be an alternative. There is also immense interest in the Mediterranean diet, which is acceptable to most patients, may decrease some biomarkers of coronary atherosclerosis, and may decrease cardiovascular events and death. Despite these options, there is no "fits all" dietary recommendation for prevention of coronary heart disease. Importantly, dietary discretion is only 1 part of lifestyle changes, such as exercise and smoking cessation. PMID- 15541242 TI - Subliminal editorial conundra confounding publication of multicenter trials. PMID- 15541243 TI - Relation of left ventricular sphericity to 10-year survival after acute myocardial infarction. AB - After ST-elevation myocardial infarction, the association between left ventricular sphericity (measured by biplane ventriculography) and survival rate at a median of 6.5 years was determined in 825 patients. The highest tertile of sphericity (vs the lowest and middle tertiles) was associated with a decreased 10 year survival rate in patients who had anterior myocardial infarction (p = 0.002), inferior myocardial infarction (p = 0.011), Thrombolysis In Myocardial Infarction (TIMI) grade 3 flow (p = 0.005), or TIMI grade 0 to 2 flow (p = 0.001) in the infarct artery. The independent multivariate predictors of a 10-year survival rate were ejection fraction (p = 0.002), treadmill exercise duration (p = 0.004), biplane left ventricular sphericity index (p = 0.032), age (p = 0.043), and end-systolic volume index (p = 0.047), but not TIMI flow grade. PMID- 15541244 TI - Prognostic significance of early short-term measurements of heart rate variability following acute myocardial infarction. AB - This study of 164 subjects demonstrates that short-term (5-minute) recordings of heart rate variability, performed within 48 hours of admission, identify those who survive acute myocardial infarction but have an adverse prognosis. PMID- 15541245 TI - Echocardiographic predictors of prognosis after first acute myocardial infarction. AB - We prospectively studied 490 patients who had a first myocardial infarction and performed a complete 2-dimensional echocardiographic study or=40% increase in cardiac index and a >or=25% decrease in pulmonary capillary wedge pressure compared with pretreatment measurements. The primary end point was reached in one of the patients treated with dobutamine alone versus 7 patients (39%) treated with levosimendan and dobutamine combined (p = 0.008), whereas at 24 hours, the combined treatment was associated with a 0.76 +/- 0.78 L/min/m(2) (p = 0.001) mean increase in cardiac index and a 6.4 +/- 7.3 mm Hg (p = 0.002) mean decrease in pulmonary capillary wedge pressure compared with measurements obtained after 24 hours of dobutamine infusion alone. Symptoms were alleviated in all patients, and all but 3 were discharged from the hospital. PMID- 15541262 TI - Prediction of subclinical left ventricular dysfunction with strain rate imaging in patients with asymptomatic severe mitral regurgitation. AB - This study sought the ability of strain rate imaging to detect subclinical left ventricular dysfunction, as evidenced by reduced contractile reserve (CR) in 32 asymptomatic patients with isolated severe mitral regurgitation. Compared with CR patients (n = 10), CR+ patients (n = 22) had significantly higher end-systolic strain and peak systolic strain rate, but these parameters were not significantly different between CR+ patients and matched normal controls. PMID- 15541263 TI - Frequency of aortic root dilation in children with a bicuspid aortic valve. AB - Bicuspid aortic valve (BAV) is associated with aortic root dilation and dissection in adults, but the age and conditions when dilation begins are unknown. Using echocardiographic data, we compared the aortic root dimensions and valve hemodynamics of 76 children with BAV with valves derived from 41 normal children. The children with BAV had significantly larger aortic roots (p <0.001) regardless of the presence of aortic stenosis or regurgitation. PMID- 15541264 TI - Comparative roles of the atrial septal aneurysm versus patent foramen ovale in systemic embolization with inferences from neonatal studies. AB - Patent foramen ovale (PFO) and atrial septal aneurysm (ASA) have a significant statistical relation to ischemic strokes in adults. We reviewed their prevalence in 106 echocardiograms of neonates. Although all normal neonates have PFO, only 4% had ASA. From fetal and neonatal echocardiograms, we conclude that ASAs are more likely to be caused by inadequate size of the PFO, but the ASA can act as a funnel, enhancing the chances of paradoxic embolization in the presence of a PFO. There is no convincing evidence that an ASA alone causes strokes in adults; the confounder is probably atrial fibrillation. PMID- 15541265 TI - Usefulness of three-dimensional echocardiographic reconstruction of the Amplatzer septal occluder in patients undergoing atrial septal closure. AB - Eighty-eight patients referred for transcatheter closure of atrial septal or patent foramen ovale defects underwent 3-dimensional transesophageal echocardiography to correlate preclosure anatomy with the morphology and positioning of the atrial septal occluder. Despite the effectiveness of the trancatheter closure and absence of complications, 2 linear indentations of the aortic root by the 2 discs of the device were clearly demonstrated in 16 cases, suggesting caution in the choice of very large occluders. PMID- 15541266 TI - Diagnosis and management of restricted or closed foramen ovale in fetuses with congenital heart disease. AB - The incidence of foramen ovale restriction or closure is greater in fetuses with congenital heart defects (CHDs) and obligate atrial shunting and/or left atrial hypertension. Foramen ovale restriction or closure in fetuses with CHD can cause distress in utero or at birth. We suggest performing serial ultrasound assessment for at-risk fetuses with CHD to best assure fetal well-being plan the timing and mode of delivery, and develop management protocols for in utero and postnatal care. The future for these patients may be more optimistic as fetal intervention procedures are developed and mastered. PMID- 15541267 TI - Initial experience with cardiac resynchronization therapy for ventricular dysfunction in young patients with surgically operated congenital heart disease. AB - Data regarding cardiac resynchronization therapy (CRT) in pediatric patients are limited. The first reported use of CRT in a young patient with congenital heart disease showed promising early results with improvement in New York Heart Association classification, aerobic capacity, and ventricular contractility 1 month after implant. Other reports have shown acute benefits of CRT in young patients with postoperative right bundle branch block after surgery for congenital heart defects. To date, however, there are no published data regarding CRT for the long-term management of systemic ventricular dysfunction in the young with associated congenital heart disease. This report describes our initial experience with CRT in these patients. PMID- 15541268 TI - Extramural coronary arteries in adults with cyanotic congenital heart disease. AB - Dilatation and tortuosity of extramural coronary arteries are prevalent in cyanotic congenital heart disease. Two pathogenetic variables are operative, namely endothelial vasodilator substances and medial structural abnormalities. PMID- 15541269 TI - Prevalence of coronary artery disease, lower extremity peripheral arterial disease, and cerebrovascular disease in 110 men with an abdominal aortic aneurysm. AB - Of 110 men (mean age 66 years) with an abdominal aortic aneurysm, 78 (71%) had coronary artery disease (CAD), 50 (46%) had lower extremity peripheral arterial disease (PAD), and 30 (27%) had cerebrovascular disease. Twenty-four percent of the patients with an abdominal aortic aneurysm had CAD plus lower extremity PAD plus cerebrovascular disease, 22% had CAD plus lower extremity PAD and no cerebrovascular disease, 22% had only CAD, 4% had CAD plus cerebrovascular disease and no lower extremity PAD, and 29% had no CAD, lower extremity PAD, or cerebrovascular disease. PMID- 15541270 TI - Safety and efficacy of citrus aurantium for weight loss. AB - To examine the safety and efficacy of citrus aurantium, an herb now commonly used as a substitute for ephedra in dietary supplements marketed to promote weight loss, we conducted a systematic review. An extensive search of MEDLINE, EMBASE, BIOSIS, and the Cochrane Collaboration Database identified only 1 eligible randomized placebo controlled trial, which followed 20 patients for 6 weeks, demonstrated no statistically significant benefit for weight loss, and provided limited information about the safety of the herb. PMID- 15541271 TI - Over-the-counter statin drug. PMID- 15541272 TI - Levels of natural IgM antibodies against phosphorylcholine in healthy individuals and in patients undergoing isolated limb perfusion. AB - Natural IgM antibodies against phosphorylcholine (anti-Pc IgM) resemble C reactive protein (CRP) regarding specificity and have gained increasing attention because of their supposed role in clearance of damaged cells and in cardiovascular disease. In order to quantify these antibodies in human plasma, we have developed an ELISA system, in which p-aminophenylphosphorylcholine (PCH) coupled to human serum albumin (HSA) was coated on microtiters plates. Human plasma or serum samples were incubated in the plates, after which bound anti-Pc IgM was detected with mouse anti-human IgM-HRP. Pre-incubation of plasma with competitors such as phosphorylcholine, phosphorylethanolamine, phosphorylserine or glycine-HSA, confirmed that the ELISA was specific for anti PC IgM. Levels of anti Pc IgM in a cohort of healthy donors differed by more than 100-fold, whereas the fluctuation of anti-Pc IgM levels in individuals over time was small (coefficient of variation between 6% to 25%). Furthermore, there was no correlation between CRP and anti-Pc IgM in this cohort. Levels of anti-Pc IgM in the normal donors correlated significantly with IgM binding to apoptotic cells. To test the hypothesis that anti-Pc IgM can bind to neo-antigens expressed on necrotic or apoptotic cells, anti-Pc IgM was also quantified in patients with tumors undergoing isolated limb perfusion with tumor necrosis factor-alpha (TNF alpha). Following this procedure, a significant decrease of circulating anti-Pc IgM relative to total IgM was found in all five patients tested. In conclusion, we have developed a specific and reproducible ELISA for anti Pc IgM quantification. Fluctuation of levels of these natural antibodies over time in healthy individuals was limited, although the variation among individuals was large. Significant decreases of levels of anti-Pc IgM were found to occur during tissue damage. PMID- 15541273 TI - A subtractive fluorescence-activated cell-sorting strategy to identify mimotopes of HBV-preS protein from bacterially displayed peptide library. AB - A novel subtractive fluorescence-activated cell-sorting (FACS) strategy using a model system is described here to identify disease-specific (DS) epitopes from a bacterially displayed random peptide library. In this process, preimmune serum was used as "Driver " to block any common binding sites on the bacterial surface and the labeled anti-preS IgG polyclonal antibodies from immunized serum were used as "Tester" to enrich preS-specific mimotopes. Bacterial clones were identified out of this pool through an "antigen-independent" procedure only using both different sera samples. After four rounds of sub-FACS screening, 41 out of 50 bacterial clones were identified as reacting with the immunized serum but not reacting with the pre-immune one. Two motif sequences HQLD and DPAF were obtained from 13 clones. Immunization of mice with two representative bacterial clones elicited a strong specific response against native preS antigen in comparison with the control. This technique may provide a useful technology platform for high-throughput screening of disease-related epitope which is of importance to develop vaccine against some infectious diseases whose pathogen or immunodominant antigen is still unknown. PMID- 15541274 TI - Chemokine networks and in vivo T-lymphocyte trafficking in nonhuman primates. AB - T-lymphocyte migratory circuits in human and nonhuman primates remain largely unexplored due to the difficulty of defining cell trafficking in vivo. However, this knowledge may reveal critical aspects of immunity and T-lymphocyte homeostasis in both health and disease. Furthermore, in vivo T-lymphocyte trafficking studies may facilitate defining mechanism(s) of immune dysfunction in the nonhuman primate model for acquired immunodeficiency syndrome (AIDS). Here, we developed a model for in vivo T-lymphocyte trafficking in nonhuman primates, and delineated homing characteristics of unstimulated peripheral blood mononuclear cells (PBMCs) to lymphoid and nonlymphoid compartments in healthy rhesus macaques. T-lymphocyte homing of autologous, carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled PBMCs was defined within 48 h of intravenous transfer. The highest relative frequency of CFSE+ T lymphocytes was observed in peripheral blood and spleen. Expression of chemokine receptor CCR7 and its ligands correlated with recirculation of T lymphocytes through the periphery and homing to paracortical regions of lymph node, where cells remained largely excluded from B-cell follicles. T-lymphocyte trafficking was also detected to the liver and bone marrow, and at low levels to the thymus and small intestine. The liver contained the highest proportion of CD45RA- T lymphocytes, consistent with homing of activated/memory T lymphocytes to this nonlymphoid site. Our data suggest that lymphoid and nonlymphoid organs are under continuous immunosurveillance in healthy macaques, and that this model may serve to investigate aberrant patterns in disease. PMID- 15541275 TI - Heat-mediated enzyme-linked immunosorbent assay procedure on a photoactivated surface. AB - The human burden of infectious and allergic disease has necessitated rapid screening and measurement of antibodies in sera. Here, we report an enhancement of speed and sensitivity of an enzyme-linked immunosorbent assay (ELISA) technique by performing it at elevated temperature on an activated surface. The activated polymer was able to bind covalently anti-human IgG at 50 degrees C over 40 min to form a solid phase. The covalently bound solid phase was stable enough to withstand subsequent ELISA steps at elevated temperature. Thus, when blocking, human IgG and antibody-enzyme conjugate binding were performed on this solid phase at 40 degrees C in 40 min, 50 degrees C in 45 min and 50 degrees C in 40 min, respectively. The ELISA readings obtained were 1.5-fold higher than those obtained at 37 degrees C over similar incubation times. Total IgE was also determined by the heat-mediated ELISA (HELISA) technique in less than 3 h and gave similar ELISA values to those obtained by the conventional procedure carried out for 18 h on an untreated surface. A stable covalently bound solid phase is a prerequisite for the HELISA technique and was further verified when a solid phase prepared through adsorption onto an untreated surface showed less than half the ELISA absorbance values obtained with the activated surface at elevated temperatures. As surface activation can be achieved by application of a simple technique, the HELISA procedure could be a powerful alternative to conventional ELISA. PMID- 15541276 TI - Towards functional transplant donor matching by measurement of granzyme A and granzyme B production levels. AB - Graft-versus-host disease (GvHD) can be a major complication after allogeneic stem cell transplantation (SCT) especially when donor and recipient are unrelated. The latter serious complication, together with the growing number of available unrelated stem cell donors, demand a simple in vitro assay for functional stem cell donor selection. Activated donor cytotoxic T lymphocytes (CTLs) and natural killer cells produce granzymes (Gr) that are involved in the pathogenesis of GvHD. We measured granzymes A and B (GrA and GrB) production levels in the supernatants of 96 h pretransplant mixed lymphocyte cultures (MLC) of 26 sibling and 31 unrelated patient/donor pairs by enzyme-linked immunosorbent assay (ELISA). In detail, the GrA and GrB production levels from a selected cohort of 37 potential patient/donor pairs were correlated with relative responses (RR) of MLC and with human leukocyte antigen (HLA) class II mismatches and with the development of acute GvHD in a second, consecutive cohort of 20 sibling SCT recipients. In vitro measurement of GrA and GrB production levels significantly correlated with the RR of pretransplant MLC (r=0.492, p< or =0.01 and r=0.853, p< or =0.01, respectively) and increased with the number of HLA class II mismatches between patient and donor. Pretransplant GrA production levels were significantly associated with the in vivo development of acute GvHD grades II-IV in patients transplanted with an HLA-identical sibling donor (p< or =0.001). In conclusion, in vitro GrA and GrB production levels can be measured by a quantitative and sensitive ELISA. This novel and simple method may be used for functional selection of unrelated stem cell donors and for the identification of patients who are at risk for acute GvHD grades II-IV. PMID- 15541277 TI - Use of multiple antigenic peptides related to antigenic determinants of infectious bursal disease virus (IBDV) for detection of anti-IBDV-specific antibody in ELISA--quantitative comparison with native antigen for their use in serodiagnosis. AB - Multiple antigenic peptides (MAPs) prepared for the predicted antigenic determinants on the VP2 protein of infectious bursal disease virus (IBDV) were used as antigens in enzyme-linked immunosorbent assay (ELISA)--an alternative to whole viral antigen to detect anti-IBDV antibodies in the chicken sera. Two MAPs were synthesized, which could specifically detect the anti-IBDV antibodies in serum samples by ELISA. The optimum quantity of MAP1 and MAP2 required to coat the wells of the ELISA plate was 5 ng/ml, whereas the amount of purified IBDV whole viral antigen was 500 ng/ml, indicating the high efficiency of MAPs. In this study, we mainly focused on the antigenicity of two eight-branched MAPs to detect anti-IBDV antibodies in ELISA, which would serve as safe, chemically defined, noninfectious alternative antigens to whole virus in serodiagnosis. The specificity and sensitivity of both MAP1 and MAP2 were found to be relatively better than the whole viral antigen. PMID- 15541278 TI - Light-chain shuffling results in successful phage display selection of functional prokaryotic-expressed antibody fragments to N-glycolyl GM3 ganglioside. AB - Phage display technology makes it possible to introduce and rapidly screen diversity in antibody binding sites. Chain shuffling has been successfully used to humanize murine antibody fragments and also to obtain affinity matured variants. Here we report a different application of this method: the use of chain shuffling to overcome improper prokaryotic expression behavior of a hybridoma derived single-chain antibody fragment. Construction and expression of such recombinant antibody fragments remain as empirical entities, hampered by the inability to express some antibody genes coming from eukaryotic cells in bacterial expression systems. Such problems are different for each combination of variable regions and can be serious enough to preclude the use of some hybridomas as sources of V regions to obtain recombinant antibody fragments. The particular binding properties and potential usefulness of some monoclonal antibodies make it highly desirable to bypass these technical limitations in order to develop smaller size therapeutic agents in the form of antibody fragments. The 14F7 mouse monoclonal antibody is one such attractive candidate due to its high specificity for the N-glycolyl GM3 ganglioside overexpressed in tumor cells and its ability to distinguish this antigen from closely related gangliosides like N-acetyl GM3. Our goal was to construct a phage-displayed single-chain Fv antibody fragment derived from 14F7. After cloning the original variable regions from the 14F7 hybridoma in a phagemid vector, we were unable to detect either binding activity or even expression of antibody fragments in bacteria, despite repetitive efforts. We constructed light-chain shuffling libraries, from which functional antibody fragments were readily selected. These combined the original 14F7 heavy chain variable region with a wide variety of unrelated murine and human light-chain variable regions. New antibody fragments retained the valuable properties of the monoclonal antibody in terms of fine specificity, affinity and tumor recognition. They were readily produced by bacteria, either in phage-displayed form or as soluble molecules, and provided a panel of potentially useful variants for cancer diagnosis and immunotherapy. Chain shuffling and phage display were found to be useful strategies for selecting antibody fragments on the basis of both prokaryotic expression and antigen binding criteria. PMID- 15541279 TI - An improved method for utilization of peptide substrates for antibody characterization and enzymatic assays. AB - Synthetic peptides have become an important tool in antibody production and enzyme characterization. The small size of peptides, however, has hindered their use in assays systems, such as Western blots, and as immunogens. Here, we present a facile method to improve the properties of peptides for multiple applications by ligating the peptides to intein-generated carrier proteins. The stoichiometric ligation of peptide and carrier achieved by intein-mediated protein ligation (IPL) results in the ligation product migrating as a single band on a SDS-PAGE gel. The carrier proteins, HhaI methylase (M.HhaI) and maltose-binding protein (MBP), were ligated to various peptides; the ligated carrier-peptide products gave sharp, reproducible bands when used as positive controls for antibodies raised against the same peptides during Western blot analysis. We further show that ligation of the peptide antigens to a different thioester-tagged carrier protein, paramyosin, produced immunogens for the production of antisera in rabbits or mice. Furthermore, we demonstrate the generation of a substrate for enzymatic assays by ligating a peptide containing the phosphorylation site for Abl protein tyrosine kinase to a carrier protein. This carrier-peptide protein was used as a kinase substrate that could easily be tested for phosphorylation using a phosphotyrosine antibody in Western blot analysis. These techniques do not require sophisticated equipment, reagents, or skills thereby providing a simple method for research and development. PMID- 15541280 TI - Enzyme-linked immunomagnetic chemiluminescent detection of Escherichia coli O157:H7. AB - E. coli O157:H7 is a pathogenic microorganism that has been implicated in numerous cases of foodborne illnesses. A variety of rapid methods exist that show promise for the presumptive detection of this pathogen without the immediate need for incubating test samples for hours to days in microbial enrichment and culture media. In recent years, highly sensitive chemiluminescence has become a more affordable and portable detection method. Chemiluminescent detection has been coupled with the selectivity of antibodies, magnetic microparticle separation/isolation, and enzymatic signal amplification in order to develop a rapid method, termed enzyme-linked immunomagnetic chemiluminescence (ELIMCL). This work presents the application of ELIMCL to the detection of E. coli O157:H7 in pristine buffered saline with a detection limit of 7.6 x 10(3) for live cells in approx. 75 min assay time. The blocking agent casein and the surfactant Tween 20 were used to lower background luminescence and thus maximize signal-to-noise ratios. After 5.5 h of enrichment culture, ELIMCL was demonstrated to detect E. coli O157:H7 inoculated in ground beef at 10 CFU/g in a total assay time of about 7 h. PMID- 15541281 TI - A novel vector for the expression of SCR domains in insect cells. AB - Exploitation of recombinant technology to study proteins containing strings of short consensus repeat (SCR) domains largely depends on expression vectors. In this paper, we describe a vector for cloning and constitutive expression of single or multiple SCR domains. The recombinant vector has unique additive features over commercially available vectors that make it a universal cloning vector for SCR domains as well as a vector suitable for expressing any protein fragment beginning and ending with cysteine residues. As a demonstration of its usefulness, the constitutive extracellular expression of five SCR-containing proteins derived from complement factor H is presented. PMID- 15541282 TI - Development and validation of a cell-based bioassay for the detection of neutralizing antibodies against recombinant human erythropoietin in clinical studies. AB - An in vitro cell-based bioassay capable of detecting neutralizing antibodies (NAb) to recombinant human erythropoietin (rHuEPO) in clinical samples was developed and validated. The bioassay uses the IL-3-dependent murine 32D cell line transfected with human EPO receptors (EPOR). This cell line responds to rHuEPO with proliferation measurable by [methyl-3H] thymidine incorporation into the cellular DNA. The reduction of rHuEPO-induced cell proliferation response indicates the possible presence of anti-rHuEPO NAb. In addition, a specificity assay using murine IL-3 (mIL-3) induced proliferation of the same cell line was developed and validated. The specificity assay allowed testing of samples that inhibited the biologic activity of rHuEPO to evaluate whether the inhibition was specific and not attributable to cytotoxicity of the serum sample. Both assays are conducted in a 5% human serum matrix in 96-well microtiter plates. The Guidelines of the International Conference on Harmonization (ICH) were followed for the validation of different assay parameters including analytical recovery, precision, sensitivity, specificity, selectivity, and robustness. The anti-rHuEPO NAb assay is capable of detecting concentrations of NAb equivalent to 500 ng/ml of the positive control antibody in undiluted human serum. The anti-rHuEPO NAb assay yielded consistent results with cells cultured for up to 30 days. The positive control antibody maintained its ability to inhibit the biologic activity of rHuEPO upon freezing and thawing. The presence of free rHuEPO in serum samples interfered with the detection of the antibody. The validated assay was sensitive, specific and robust and was successfully used to monitor NAb development in patients. PMID- 15541283 TI - Comparative analysis of lymphocyte activation marker expression and cytokine secretion profile in stimulated human peripheral blood mononuclear cell cultures: an in vitro model to monitor cellular immune function. AB - Activation of lymphocytes is a complex, yet finely regulated cascade of events that results in the expression of cytokine receptors, production and secretion of cytokines and expression of several cell surface molecules that eventually lead to divergent immune responses. Assessing the qualitative and quantitative nature of lymphocyte function following immunotherapy provides valuable information about the immune responses mediated by a therapeutic agent. To facilitate evaluation of the immunomodulatory activity of therapeutic agents, we have established a platform of in vitro immunoassays with normal human peripheral blood mononuclear cells (PBMCs) treated with several polyclonal activators that are known to exhibit different modes of action. We evaluated the kinetics of cell surface marker expression and cytokine release from PBMCs stimulated in parallel with various activating agents over a time course. These stimulating agents induced early (CD69 and CD71) and late (CD25 and HLA-DR) activation markers to varying antigen densities, indicated different cytokine profiles, and showed differential inhibition with dexamethasone (DEX), an inhibitor of early signaling events. Based on the association or correlation of the kinetics of activation marker expression and secreted cytokines, the results of our study indicate the appropriate time points for the simultaneous measurement of both these activation products. This study defines the kinetics for both measures of T cell activation and provides a comprehensive review with various polyclonal activators that can serve as a reference for monitoring lymphocyte function in clinical study samples. PMID- 15541284 TI - A new immunofluorostaining method using red fluorescence of PerCP on formalin fixed paraffin-embedded tissues. AB - Immunofluorostaining, a versatile tissue staining method, is used in biomedical research because of its clear contrast and precise quantification of positive signals. However, its application in clinical diagnosis has been limited. A major obstacle is high fluorescent background of formalin-fixed, paraffin-embedded tissue sections (paraffin sections). On paraffin sections, strong and broad fluorescence of the section overlapping that of conventional fluorescent dyes such as fluorescein isothiocyanate (FITC) prevents detection of target immunofluorescence. To circumvent the background, we selected an albuminous dye, peridinin chlorophyll a protein (PerCP), for immunostaining of human tumor sections with tumor-reactive monoclonal antibodies. Red fluorescence of PerCP clearly distinguished the tumor region within the yellow-green autofluorescence of the section. Furthermore, it was possible to observe tissue morphology simultaneously without any counterstaining; autofluorescence served as counterstaining in this method. Digital quantification of PerCP-stained image intensity correlated (r2>0.99) well with extracted PerCP amount, indicating the usefulness of image quantification. We conclude that this new and simple immunofluorostaining method can be applied to pathological diagnosis of a wide range of conditions, including cancer. PMID- 15541285 TI - Long-term collection and characterization of afferent lymph from the ovine small intestine. AB - Reliable methods for long-term collection of afferent lymph draining from the small intestine of sheep are described and validated. The procedure was used successfully in normal sheep, in animals infected experimentally with the parasitic intestinal nematode Trichostrongylus colubriformis and in animals infected naturally with Mycobacterium avium subsp. paratuberculosis, the causative agent of Johne's disease. Our approach enabled afferent lymph draining from the small intestine to be collected continuously for up to 4 months, without any detrimental effects on the animals. Based on cytokine gene expression profiles of afferent intestinal lymph cells, the two infections induced contrasting regional immune responses, namely, Th2-type immunity in the case of T. colubriformis infection and Th1-type immunity in natural cases of Johne's disease. Some immune parameters differed markedly between the two disease models, highlighting the potential value of this approach to gain real-time insights into distinctive host-pathogen interactions as they occur in vivo within the regional immune system of the gastrointestinal tract. PMID- 15541286 TI - Implication of delayed TNF-alpha exposure on dendritic cell maturation and expansion from cryopreserved cord blood CD34+ hematopoietic progenitors. AB - Most currently used systems for dendritic cell (DC) production from progenitors entail tumor necrosis factor alpha (TNF-alpha) at the onset of cell culture, based on the notion that TNF-alpha might be required in the early stages of DC development. To optimize conditions for DC expansion from cryopreserved cord blood (CB) CD34+ hematopoietic progenitors, we took a dynamic approach to define the timing of TNF-alpha exposure to the culture. We cultured cord blood CD34+ cells in RPMI-1640 with 10% human AB plasma, stem cell factor (days 1-6), granulocyte-macrophage colony-stimulating factor (days 1-18), interleukin-4 (days 6-18) and varying schedules of TNF-alpha (0-144 h after thawing). Expression of the DC-associated markers, including CD83/CD1a, HLA DR/CD86/CD80, CD14/CD40, was monitored every 3 days. Our data demonstrate that delayed TNF-alpha exposure by 48-72 h after thawing gave rise to two- to three-fold increase in the yield of CD83+ DCs that were highly active in stimulating allogeneic T-cell proliferation compared to immediate TNF-alpha exposure. Thus, the immediate exposure of cryopreserved cord blood CD34+ cells to TNF-alpha, potentially compromising DC expansion, should be avoided. This finding should be of significant consideration when using cryopreserved CD34+ progenitor cells as a source of immunologically competent DCs in a clinical setting. PMID- 15541287 TI - ELISAs for free human immunoglobulin light chains in serum: improvement of assay specificity by using two specific antibodies in a sandwich detection method. AB - Free forms of light chains (FLCs) in serum are clinically useful markers, but their practical use in laboratories has been limited due to the cross-reaction with light chains associated with heavy chains (intact immunoglobulins) on FLC assays. In this study, we attempted to use anti-FLC antiserum and anti-FLC monoclonal antibody (mAb) in a sandwich enzyme-linked immunosorbent assay (ELISA) for serum FLCs. This combination resulted in a synergistic effect, showing 10(5) fold specificity for FLC kappa and 10(4)-fold for FLC lambda compared to IgG. The specific ELISAs had good sensitivity, detecting 0.78 microg/l of FLC kappa and 31.3 microg/l of FLC lambda. We then measured serum samples before and after the absorption of IgG, IgA, and IgM and found that there was no appreciable difference in the FLC values between them. These results indicate that the ELISAs quantify serum FLCs more precisely even in the presence of about a thousand-fold amount of intact immunoglobulin. Meanwhile, the use of two anti-FLC mAbs in an ELISA showed no response for FLCs probably because of the conflict of the two mAbs at the specific epitope in FLC. Thus, ELISAs with anti-FLC antiserum and anti-FLC mAb can provide a reliable method for the clinical test of serum FLCs. PMID- 15541288 TI - A single monoclonal antibody as probe to detect the entire set of native and partially unfolded rhEPO glycoforms. AB - Human erythropoietin (hEPO) is a highly heterogeneous glycosylated protein that requires well-characterised immunochemical reagents to evaluate the glycoform profile along its biotechnological production as a recombinant hormone. These reagents should be suitable for several assay conditions (like those used for immunoblotting analysis, liquid or solid-phase quantitative assays, immunoaffinity purification) with no glycoform selectivity. Five anti-recombinant hEPO monoclonal antibodies (mAbs) were characterised with the aim of selecting the appropriate reagent. These antibodies mapped two spatially distinct epitopes and neutralised the in vitro biological activity of the cytokine. All of them were able to bind to both, the partially denatured and the native form of the protein. Isoelectric focusing analysis followed by immunoblotting confirmed that all the mAbs, herein described, were able to bind to each glycoform, recognising amino acid sequences of the hEPO. Nevertheless, only mAb 2B2 preserved the ability to bind to soluble recombinant human erythropoietin (rhEPO) when it was coated to polystyrene plates or immobilised on CNBr-activated Sepharose matrix. Besides, mAb 2B2 was able to bind to the complete set of soluble rhEPO glycoforms, showing the same affinity for the glycosylated and deglycosylated cytokine. Thus, mAb 2B2 was useful as a capture antibody to develop a sandwich enzyme-linked immunosorbent assay (ELISA), performing a simple, specific and fast assay to quantify rhEPO with a detection limit of 7 ng ml(-1). mAb 2B2 was also satisfactorily employed as affinity ligand to purify rhEPO. Our work led us to find a suitable and single reagent to perform a variety of immunochemical approaches, where the binding of each glycoform in the native or partially unfolded form of rhEPO is required. PMID- 15541289 TI - An enzyme-linked immunosorbent assay for human carcinoembryonic antigen-related cell adhesion molecule 8, a biological marker of granulocyte activities in vivo. AB - Carcinoembryonic antigen-related cell adhesion molecule 8 (CEACAM8), also known as CD66b, NCA-95 and CD67, is a highly glycosylated protein expressed only in neutrophils and eosinophils in humans. The precise function of CEACAM8 remains unclear. As a member of the family of carcinoembryonic antigen (CEA), it may play a role in the interaction between granulocytes or between granulocytes and epithelial cells. We describe here an accurate, specific and reproducible enzyme linked immunosorbent assay (ELISA) using purified native CEACAM8 as standard for the measurement of CEACAM8 with a detection range of 1-64 microg/l. We also present data on the levels of CEACAM8 in the blood of healthy individuals and patients undergoing surgery, as well as in patients with acute infections. The highly elevated levels of CEACAM8 in the blood of these patients, which are significantly correlated with the surface expression of CEACAM8 on neutrophils and the number of circulating neutrophils, suggest that CEACAM8 could serve as a biological marker for granulocyte activities in vivo. PMID- 15541290 TI - Isolating pure populations of monocytes from the blood of pregnant women: comparison of flotation in iodixanol with elutriation. AB - Observations that the innate arm of the immune system is upregulated in pregnancy have highlighted the need for methods of isolating pure populations of monocytes for studies into pregnancy and pre-eclampsia without activating them during the isolation process. Density gradient centrifugation using iodixanol is a useful method for isolating relatively pure populations of unactivated monocytes from human blood but has not been validated in pregnant subjects. We compared the ability of monocytes isolated from pregnant women by density gradient centrifugation using iodixanol (n=6) with monocytes isolated by countercurrent centrifugal elutriation (n=6) in terms of their ability to produce interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) under basal conditions and after stimulation with bacterial lipopolysaccharide (LPS). Under basal conditions, monocytes isolated by density gradient centrifugation produced low amounts of IL-6 and MCP-1. Production of IL-6 and MCP-1 after stimulation of the monocytes with LPS was much greater (p<0.01). There was no statistically significant difference between the two methods in terms of stimulated levels of either cytokine. PMID- 15541291 TI - Heterologous expression of a plant arginine decarboxylase gene in Trypanosoma cruzi. AB - Wild-type Trypanosoma cruzi epimastigotes lack arginine decarboxylase (ADC) enzymatic activity. However, the transformation of these parasites with a recombinant plasmid containing the oat ADC cDNA coding region gave rise to the transient heterologous expression of the enzyme, suggesting the absence of endogenous mechanisms that could inhibit the expression of a hypothetical own ADC gene or the assay used to measure its enzymatic activity. The foreign ADC enzyme expressed in the transgenic T. cruzi was characterized by identification of the products, the stoichiometry of the catalysed reaction, the specific inhibition by alpha-difluoromethylarginine (DFMA) and the study of its metabolic turnover. The half-life of the heterologous ADC activity in T. cruzi was about 150 min. Bioinformatics studies and polymerase chain reaction (PCR) analyses seem to indicate the absence of ADC-like DNA sequences in the wild-type T. cruzi genome. PMID- 15541292 TI - Fifty-hertz magnetic fields induce free radical formation in mouse bone marrow derived promonocytes and macrophages. AB - Our findings show a significant increase of free radical production after exposure to 50 Hz electromagnetic fields at a flux density of 1 mT to mouse bone marrow-derived (MBM) promonocytes and macrophages, indicating the cell-activating capacity of extremely low frequency magnetic fields (ELF-MF). We demonstrate that after exposure to ELF-MF mainly superoxide anion radicals were produced, both in MBM macrophages (33%) and also in their precursor cells (24%). To elucidate whether NADPH- or NADH-oxidase functions are target proteins for MF interaction, the flavoprotein inhibitor diphenyleneiodonium chloride (DPI) was used. MF induced free radical production was not inhibited by DPI, whereas tetradecanoylphorbolacetate (TPA)-induced free radical production was diminished by about 70%. TPA is known to induce a direct activation of NADPH-oxidase through the PKC pathway. Since DPI lacks an inhibitory effect in MF-exposed MBM cells, we suggest that 50 Hz MF stimulates the NADH-oxidase pathway to produce superoxide anion radicals, but not the NADPH pathway. Furthermore, we showed an oscillation (1-10 days) in superoxide anion radical release in mouse macrophages, indicating a cyclic pattern of NADH-oxidase activity. PMID- 15541293 TI - Purification and characterization of two forms of endo-beta-1,4-mannanase from a thermotolerant fungus, Aspergillus fumigatus IMI 385708 (formerly Thermomyces lanuginosus IMI 158749). AB - Two extracellular endo-beta-1,4-mannanases, MAN I (major form) and MAN II (minor form), were purified to electrophoretic homogeneity from a locust bean gum-spent culture fluid of Aspergillus fumigatus IMI 385708 (formerly Thermomyces lanuginosus IMI 158749). Molecular weights of MAN I and MAN II estimated by SDS PAGE were 60 and 63 kDa, respectively. IEF afforded several glycoprotein bands with pI values in the range of 4.9-5.2 for MAN I and 4.75-4.9 for MAN II, each exhibiting enzyme activity. MAN I as well as MAN II showed highest activity at pH 4.5 and 60 degrees C and were stable in the pH range 4.5-8.5 and up to 55 degrees C. In accordance with the ability of the enzymes to catalyze transglycosylation reactions, 1H NMR spectroscopy of reaction products generated from mannopentaitol confirmed the retaining character of both enzymes. Both MAN I and MAN II exhibited essentially identical kinetic parameters for polysaccharides and a similar hydrolysis pattern of various oligomeric and polymeric substrates. Both beta-mannanases contained identical internal amino acid sequence corresponding to glycoside hydrolase family 5 and also a cellulose-binding module. These data suggested that both MAN I and MAN II are products of the same gene differing in posttranslational modification. Indeed, the corresponding gene was identified within the recently sequenced Aspergillus fumigatus genome (http://sanger.ac.uk/Projects/A_fumigatus/). PMID- 15541294 TI - Do the serum oxidative stress biomarkers provide a reasonable index of the general oxidative stress status? AB - The oxidant status of an individual is assessed by determining a group of markers in noninvasive samples. One limitation when measuring these biomarkers is that they do not give information about tissue localization of oxidative stress. The present study was undertaken to establish whether the serum oxidative stress biomarkers are indicative of oxidative stress in tissues of an individual. To accomplish this, we determined a few generic markers of oxidation in serum and tissues of six groups of rats treated experimentally, to modulate their oxidative stress status. The correlation between serum and tissue levels was calculated for each marker. Also, for each tissue, the correlation between the values of these oxidative stress biomarkers was analysed. Our results show that only lipid peroxides in serum could be useful to predict the oxidative stress in tissues. No correlation was found between any of the oxidative stress markers in serum. PMID- 15541295 TI - Iron nitrosyl hemoglobin formation from the reaction of hydroxylamine and hemoglobin under physiological conditions. AB - Sickle cell disease patients receiving hydroxyurea (HU) therapy have shown increases in the production of nitric oxide (NO) metabolites, which include iron nitrosyl hemoglobin (HbNO), nitrite, and nitrate. However, the exact mechanism by which HU forms HbNO in vivo is not understood. Previous studies indicate that the reaction of oxyhemoglobin (oxyHb) or deoxyhemoglobin (deoxyHb) with HU are too slow to account for in vivo HbNO production. In this study, we show that the reaction of methemoglobin (metHb) with HU to form HbNO could potentially be fast enough to account for in vivo HbNO formation but competing reactions of either excess oxyHb or deoxyHb during the reaction reduces the likelihood that HbNO will be produced from the metHb-HU reaction. Using electron paramagnetic resonance (EPR) spectroscopy we have detected measurable amounts of HbNO and metHb during the reactions of oxyHb, deoxyHb, and metHb with excess hydroxylamine (HA). We also demonstrate HbNO and metHb formation from the reactions of excess oxyHb, deoxyHb, or metHb and HA, conditions that are more likely to mimic those in vivo. These results indicate that the reaction of hydroxylamine with hemoglobin produces HbNO and lend chemical support for a potential role for hydroxylamine in the in vivo metabolism of hydroxyurea. PMID- 15541296 TI - Specific xyloglucanases as a new class of polysaccharide-degrading enzymes. AB - Three specific xyloglucanases (XGs) were isolated from Aspergillus japonicus (32 kDa, pI 2.8), Chrysosporium lucknowense (78 kDa, pI 3.8) and Trichoderma reesei (75-105 kDa, pI 4.1-4.3). The characteristic feature of these enzymes was their high specific activity toward tamarind xyloglucan, whereas the activity against carboxymethylcellulose (CMC) and barley beta-glucan was absent or very low. Peptide mass fingerprinting using MALDI-TOF mass spectrometry showed that the T. reesei XG represents Cel74A, whose gene has been discovered recently (GenBank accession no. AY281371 ), but the enzyme has not been characterized and described elsewhere. Tryptic peptides from A. japonicus and C. lucknowense xyloglucanases did not show any identity to those from known glycoside hydrolases. All enzymes produced XXXG, XXLG/XLXG and XLLG oligosaccharides as the end products of xyloglucan hydrolysis. A. japonicus XG displayed an endo-type of attack on the polymeric substrate, while the mode of action of two other xyloglucanases was similar to the exo-type, when oligosaccharides containing four glucose residues in the main chain were split off the ends of xyloglucan molecules. These results together with growing literature data allow concluding that specific xyloglucanases may represent a new class of glycoside hydrolases, which are different from regular endo-1,4-beta-glucanases. PMID- 15541297 TI - Specificity of Amaranthus leucocarpus syn. hypocondriacus lectin for O glycopeptides. AB - Amaranthus leucocarpus syn. hypochondriacus lectin (ALL) has been shown to be specific for N-acetyl-D-galactosamine (GalNAc). In this work, we determined a value of 1.0 x 10(-2) M for the association constant of ALL for GalNAc, calculated using fluorescence spectroscopy assays. Using neoglycopeptides obtained by in vitro O-glycosylation, we determined the main features of O glycopeptides recognized by ALL using molecular dynamics simulations, capillary electrophoresis, and ELISA. Neo-glycopeptides were obtained by in vitro O glycosylation reaction using microsomal preparations of murine thymocytes, human gastric fundus and colonic mucosa. ELISA assays were performed with peroxidase labeled murine monoclonal IgG2, kappa light chain (5D4) antibodies against ALL. Among the in vitro neoglycopeptides, only those of TTSAPTTS containing GalNAc at Thr in #2 and #6 reacted with ALL. Neither the TTSAPTTS glycopeptide, containing a unique GalNAc residue at Thr in #2, nor others (with more than two GalNAc residues) interacted with the lectin. Computational docking assays of the lower energy conformers for interactions between glycopeptides and lectins confirmed that ALL recognized GalNAc residues when they are spaced out in glycan structures, whereas GalNAc residues arranged in clusters prevented interaction with the lectin, indicating that ALL is specific for a special GalNAc-containing motif found in different O-glycoproteins. PMID- 15541298 TI - The structure-activity relationship of various YO compounds, novel plasmin inhibitors, in the apoptosis induction. AB - We have previously reported that YO-2, a selective plasmin inhibitor, induces thymocyte apoptosis. To elucidate the mechanism of YO-2-induced apoptosis, other YO compounds with different plasmin inhibitory action were tested for the pro apoptotic activity in this study. The treatment of rat thymocytes with the YO compounds which had the hydrophobic but not the hydrophilic moiety at the C terminal increased DNA fragmentation, the number of condensed nuclei and caspase 3-like activity. All pro-apoptotic YO compounds not only were potent plasmin inhibitors but also had the hydrophobic C-terminal as the common structure. Therefore, the target molecule of the YO compounds may be located not on the cell surface but rather inside the cells. PMID- 15541299 TI - Molecular association of lectin and beta-glucosidase in corn coleoptile. AB - Corn coleoptile lectin is present with beta-glucosidase (EC. 3.2.1.2.1) in a single tightly bound molecular association complex (88.7 kDa). SDS-PAGE of the molecular complex dissociates into two main components. Of these, at a concentration of 75%, the corn coleoptile beta-glucosidase (60 kDa) is identified by enzymatic activity, with two 16-amino acid tryptic peptides displaying close homology with the primary structure of the enzyme. In separate experiments, we isolated homogenous monomeric enzyme of corn coleoptile. This allowed us to conclude that lectin properties like erythrocyte agglutination, found in the (88.7 kDa) molecular complex, is not due to the beta-glucosidase bound in it. Another protein (30 kDa) dissociated from the same SDS-PAGE gels rendered several tryptic peptides, including a 20-amino acid sequence V(L)GP(Q)W(A)GGSGGSPVDITAEPQR closely homologous to the putative beta-glucosidase aggregating factor (BGAF) precursor described recently. Tryptic peptide SAFTE(A)WN(V)ELK(V) was also present in the BGAF precursor. KFHEQR peptide was not present in BGAF precursor or any other protein sequence examined. Tryptic peptide TYGPFGA showed good homology with the BGAF precursor protein, FEGLYLFHTPLGSGAN peptide displayed identity with the BGAF precursor sequence. Thus, the 30 kDa protein does not appear to be identical to BGAF, but is rather a similar molecule which could be endowed with the lectin properties of the 88.7 kDa molecular complex. PMID- 15541300 TI - Exaggerated polymerisation of beta-amyloid 40 stimulated by plasma lipoproteins results in fibrillar Abeta preparations that are ineffective in promoting ADP induced platelet aggregation. AB - The cytotoxic beta-amyloid peptide (Abeta) of Alzheimer's disease (AD) occurs in both plasma and platelets and may modulate platelet function. Its biological activity may relate to its fibril content and factors that promote Abeta fibrillogenesis, e.g., plasma lipoproteins could, therefore, have implications for Abeta action. We undertook a study in which structure-activity relationships were considered with respect to the actions of Abeta(1-40) on platelet function. Thus, the influence of soluble Abeta and various fibrillar Abeta preparations (0.1-10 microM) on platelet aggregation and endogenous 5-hydroxytryptamine (5-HT) efflux was investigated. Soluble Abeta(1-40) only enhanced platelet aggregation (+30%, P<0.05) and 5-HT release (+28%) stimulated by ADP (1 microM) at the highest concentration tested (10 microM). By contrast, fibrillar Abeta(1-40) at 1, 5 and 10 microM potentiated aggregation by 17.4%, 68.8% (P<0.05) and 99.5% (P<0.0001), respectively, and 5-HT efflux by 17.4%, 65% and 208% (P<0.001). Abeta(1-40) fibrils generated in the presence of native and oxidised very low density lipoprotein (VLDL), low-density lipoprotein (LDL) and high-density lipoprotein (HDL) yielded platelet responses that did not differ from those seen with the lipoproteins alone. These responses were markedly lower than those obtained with homogeneous Abeta fibrils. Our data indicate that homogeneous Abeta(1-40) fibrils are more potent than soluble Abeta(1-40) in promoting platelet reactivity and that interactions with plasma lipoproteins result in the formation of Abeta fibrils that are ineffective. We suggest that lipoproteins may interfere with the recognition of Abeta by appropriate platelet receptors and/or cause Abeta to assume an "overaggregated" biologically inert state. PMID- 15541301 TI - Inhibitory effects of trientine, a copper-chelating agent, on induction of DNA strand breaks in hepatic cells of Long-Evans Cinnamon rats. AB - Effects of treatment with trientine, a specific copper-chelating agent, on accumulation of copper and induction of DNA strand breaks were investigated in Long-Evans Cinnamon (LEC) rats, an animal model for human Wilson's disease. Copper accumulated in the livers of LEC rats in an age-dependent manner from 4 to 13 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, hepatic copper contents did not increase and were maintained at the same levels as those in 10-week-old LEC rats. When the amounts of DNA single-strand breaks (SSBs) were estimated by a comet assay, SSBs of DNA were induced in a substantial population of LEC rat hepatic cells around 8 weeks of age and the amounts of SSBs increased in an age-dependent manner from 8 to 15 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, the observed number of cells with DNA damage decreased dramatically, suggesting that induction of SSBs of DNA was inhibited and/or SSBs were repaired during the period of treatment with trientine. The results show that treatment of LEC rats with trientine decreases the number of DNA strand breaks observed, although copper contents remain high in the liver. PMID- 15541302 TI - The alpha-helical membrane spanning domain of cytochrome b5 interacts with cytochrome P450 via nonspecific interactions. AB - Cytochrome b5 (cyt b5) is an amphipathic membrane-bound heme protein found in the endoplasmic reticulum of eukaryotes. It consists of three domains, an N-terminal cytosolic, hydrophilic domain containing the heme, a short flexible linker and an alpha-helical membrane-spanning domain. This study investigated whether there are specific side chain helix-helix packing interactions between the COOH-terminal membrane anchor of cyt b5 and cytochrome P450 (cyt P450) 2B4 in a purified reconstituted system. Alanine was inserted at six positions in the membrane anchor of cyt b5. Insertion of alanine into an alpha-helix causes all amino acids at its carboxyl terminus to be rotated by 100 degrees . The ability of the alanine insertion mutants of cyt b5 to bind to cyt P450 2B4 was similar to that of the wild-type protein as was the ability of the mutant cyts b5 to stimulate the metabolism of the anesthetic, methoxyflurane. These results demonstrate that the C-terminal hydrophobic alpha-helix of cyt b5 does not interact with cyt P450 2B4 through a specific stereochemical fit of amino acid side chains, but rather through nonspecific interactions. PMID- 15541303 TI - Protein kinases linked to the pathogenesis of Parkinson's disease. AB - Two papers in this issue of Neuron identify a causative gene, LRRK2, for familial parkinsonism. Several dominantly inherited missense mutations have been identified in a number of families that exhibit a broad spectrum of neuropathological features, including deposition of alpha-synuclein and tau proteins. The LRRK2 gene is predicted to encode a large protein containing leucine-rich repeats and Ras/GTPase, tyrosine kinase-like, and WD40 domains. PMID- 15541304 TI - AMPA receptors bring on the pain. AB - The role of Ca(2+)-permeable AMPA receptors in pain processing has not been extensively studied. In this issue of Neuron, Hartmann et al. show that altering the levels of these receptors has consequences for inflammatory pain hypersensitivity but not acute pain processing. PMID- 15541305 TI - Pruning an axon piece by piece: a new mode of synapse elimination. AB - The process by which excess axons are pruned during development has remained unclear. In this issue of Neuron, Bishop et al. use time-lapse imaging and serial electron microscopy of developing neuromuscular junctions to describe a novel cellular mechanism in which retracting axon branches shed fragments rich in normal synaptic organelles. These "axosomes" are engulfed by adjacent Schwann cells and may be assimilated into the glial cytoplasm. Shedding of axosomes and glial engulfment may represent a widespread mechanism of synapse elimination. PMID- 15541306 TI - Ensemble dynamics of hippocampal regions CA3 and CA1. AB - Computational models based on hippocampal connectivity have proposed that CA3 is uniquely positioned as an autoassociative memory network, capable of performing the competing functions of pattern completion and pattern separation. Recently, three independent studies, two using parallel neurophysiological recording methods and one using immediate-early gene imaging, have examined the responses of CA3 and CA1 ensembles to alterations of environmental context in rats. The results provide converging evidence that CA3 is capable of performing nonlinear transformations of sensory input patterns, whereas CA1 may represent changes in input in a more linear fashion. PMID- 15541307 TI - A unique PDZ ligand in PKCalpha confers induction of cerebellar long-term synaptic depression. AB - Induction of cerebellar long-term depression (LTD) requires a postsynaptic cascade involving activation of mGluR1 and protein kinase C (PKC). Our understanding of this process has been limited by the fact that PKC is a large family of molecules, many isoforms of which are expressed in the relevant postsynaptic compartment, the cerebellar Purkinje cell. Here, we report that LTD is absent in Purkinje cells in which the alpha isoform of PKC has been reduced by targeted RNA interference or in cells derived from PKCalpha null mice. In both of these cases, LTD could be rescued by expression of PKCalpha but not other PKC isoforms. The special role of PKCalpha in cerebellar LTD is likely to derive from its unique PDZ ligand (QSAV). When this motif is mutated, PKCalpha no longer supports LTD. Conversely, when this PDZ ligand is inserted in a nonpermissive isoform, PKCgamma, it confers the capacity for LTD induction. PMID- 15541308 TI - Cloning of the gene containing mutations that cause PARK8-linked Parkinson's disease. AB - Parkinson's disease (PD; OMIM #168600) is the second most common neurodegenerative disorder in the Western world and presents as a progressive movement disorder. The hallmark pathological features of PD are loss of dopaminergic neurons from the substantia nigra and neuronal intracellular Lewy body inclusions. Parkinsonism is typically sporadic in nature; however, several rare familial forms are linked to genetic loci, and the identification of causal mutations has provided insight into the disease process. PARK8, identified in 2002 by Funayama and colleagues, appears to be a common cause of familial PD. We describe here the cloning of a novel gene that contains missense mutations segregating with PARK8-linked PD in five families from England and Spain. Because of the tremor observed in PD and because a number of the families are of Basque descent, we have named this protein dardarin, derived from the Basque word dardara, meaning tremor. PMID- 15541309 TI - Mutations in LRRK2 cause autosomal-dominant parkinsonism with pleomorphic pathology. AB - We have previously linked families with autosomal-dominant, late-onset parkinsonism to chromosome 12p11.2-q13.1 (PARK8). By high-resolution recombination mapping and candidate gene sequencing in 46 families, we have found six disease-segregating mutations (five missense and one putative splice site mutation) in a gene encoding a large, multifunctional protein, LRRK2 (leucine rich repeat kinase 2). It belongs to the ROCO protein family and includes a protein kinase domain of the MAPKKK class and several other major functional domains. Within affected carriers of families A and D, six post mortem diagnoses reveal brainstem dopaminergic degeneration accompanied by strikingly diverse pathologies. These include abnormalities consistent with Lewy body Parkinson's disease, diffuse Lewy body disease, nigral degeneration without distinctive histopathology, and progressive supranuclear palsy-like pathology. Clinical diagnoses of Parkinsonism with dementia or amyotrophy or both, with their associated pathologies, are also noted. Hence, LRRK2 may be central to the pathogenesis of several major neurodegenerative disorders associated with parkinsonism. PMID- 15541310 TI - Activated CREB is sufficient to overcome inhibitors in myelin and promote spinal axon regeneration in vivo. AB - Inhibitors in myelin play a major role in preventing spontaneous axonal regeneration after CNS injury. Elevation of cAMP overcomes this inhibition, in a transcription-dependent manner, through the upregulation of Arginase I (Arg I) and increased synthesis of polyamines. Here, we show that the cAMP effect requires activation of the transcription factor cAMP response element binding protein (CREB) to overcome myelin inhibitors; a dominant-negative CREB abolishes the effect, and neurons expressing a constitutively active form of CREB are not inhibited. Activation of CREB is also required for cAMP to upregulate Arg I, and the ability of constitutively active CREB to overcome inhibition is blocked by an inhibitor of polyamine synthesis. Finally, expression of constitutively active CREB in DRG neurons is sufficient to promote regeneration of subsequently lesioned dorsal column axons. These results indicate that CREB plays a central role in overcoming myelin inhibitors and so encourages regeneration in vivo. PMID- 15541311 TI - GFRalpha1 expression in cells lacking RET is dispensable for organogenesis and nerve regeneration. AB - The GDNF family ligands signal through a receptor complex composed of a ligand binding subunit, GFRalpha, and a signaling subunit, the RET tyrosine kinase. GFRalphas are expressed not only in RET-expressing cells, but also in cells lacking RET. A body of evidence suggests that RET-independent GFRalphas are important for (1) modulation of RET signaling in a non-cell-autonomous fashion (trans-signaling) and (2) regulation of NCAM function. To address the physiological significance of these roles, we generated mice specifically lacking RET-independent GFRalpha1. These mice exhibited no deficits in regions where trans-signaling has been implicated in vitro, including enteric neurons, motor neurons, kidney, and regenerating nerves. Furthermore, no abnormalities were found in the olfactory bulb, which requires proper NCAM function for its formation and is putatively a site of GDNF-GFRalpha-NCAM signaling. Thus RET independent GFRalpha1 is dispensable for organogenesis and nerve regeneration in vivo, indicating that trans-signaling and GFRalpha-dependent NCAM signaling play a minor role physiologically. PMID- 15541312 TI - The AMPA receptor subunits GluR-A and GluR-B reciprocally modulate spinal synaptic plasticity and inflammatory pain. AB - Ca(2+)-permeable AMPA receptors are densely expressed in the spinal dorsal horn, but their functional significance in pain processing is not understood. By disrupting the genes encoding GluR-A or GluR-B, we generated mice exhibiting increased or decreased numbers of Ca(2+)-permeable AMPA receptors, respectively. Here, we demonstrate that AMPA receptors are critical determinants of nociceptive plasticity and inflammatory pain. A reduction in the number of Ca(2+)-permeable AMPA receptors and density of AMPA channel currents in spinal neurons of GluR-A deficient mice is accompanied by a loss of nociceptive plasticity in vitro and a reduction in acute inflammatory hyperalgesia in vivo. In contrast, an increase in spinal Ca(2+)-permeable AMPA receptors in GluR-B-deficient mice facilitated nociceptive plasticity and enhanced long-lasting inflammatory hyperalgesia. Thus, AMPA receptors are not mere determinants of fast synaptic transmission underlying basal pain sensitivity as previously thought, but are critically involved in activity-dependent changes in synaptic processing of nociceptive inputs. PMID- 15541313 TI - Axon branch removal at developing synapses by axosome shedding. AB - In many parts of the developing nervous system, the number of axonal inputs to each postsynaptic cell is dramatically reduced. This synapse elimination has been extensively studied at the neuromuscular junction, but how axons are lost is unknown. Here, we combine time-lapse imaging of fluorescently labeled axons and serial electron microscopy to show that axons at neuromuscular junctions are removed by an unusual cellular mechanism. As axons disappear, they shed numerous membrane bound remnants. These "axosomes" contain a high density of synaptic organelles and are formed by engulfment of axon tips by Schwann cells. After this engulfment, the axosome's contents mix with the cytoplasm of the glial cell. Axosome shedding might underlie other forms of axon loss and may provide a pathway for interactions between axons and glia. PMID- 15541314 TI - The translational repressor Pumilio regulates presynaptic morphology and controls postsynaptic accumulation of translation factor eIF-4E. AB - Translational repression by Drosophila Pumilio (Pum) protein controls posterior patterning during embryonic development. Here, we show that Pum is an important mediator of synaptic growth and plasticity at the neuromuscular junction (NMJ). Pum is localized to the postsynaptic side of the NMJ in third instar larvae and is also expressed in larval neurons. Neuronal Pum regulates synaptic growth. In its absence, NMJ boutons are larger and fewer in number, while Pum overexpression increases bouton number and decreases bouton size. Postsynaptic Pum negatively regulates expression of the translation factor eIF-4E at the NMJ, and Pum binds selectively to the 3'UTR of eIF-4E mRNA. The GluRIIa glutamate receptor is upregulated in pum mutants. These results, together with genetic epistasis studies, suggest that postsynaptic Pum modulates synaptic function via direct control of eIF-4E expression. PMID- 15541315 TI - CPG2: a brain- and synapse-specific protein that regulates the endocytosis of glutamate receptors. AB - Long-term maintenance and modification of synaptic strength involve the turnover of neurotransmitter receptors. Glutamate receptors are constitutively and acutely internalized, presumptively through clathrin-mediated receptor endocytosis. Here, we show that cpg2 is a brain-specific splice variant of the syne-1 gene that encodes a protein specifically localized to a postsynaptic endocytotic zone of excitatory synapses. RNAi-mediated CPG2 knockdown increases the number of postsynaptic clathrin-coated vesicles, some of which traffic NMDA receptors, disrupts the constitutive internalization of glutamate receptors, and inhibits the activity-induced internalization of synaptic AMPA receptors. Manipulating CPG2 levels also affects dendritic spine size, further supporting a function in regulating membrane transport. Our results suggest that CPG2 is a key component of a specialized postsynaptic endocytic mechanism devoted to the internalization of synaptic proteins, including glutamate receptors. The activity dependence and distribution of cpg2 expression further suggest that it contributes to the capacity for postsynaptic plasticity inherent to excitatory synapses. PMID- 15541316 TI - Bidirectional parallel fiber plasticity in the cerebellum under climbing fiber control. AB - Cerebellar parallel fiber (PF)-Purkinje cell (PC) synapses can undergo postsynaptically expressed long-term depression (LTD) or long-term potentiation (LTP) depending on whether or not the climbing fiber (CF) input is coactivated during tetanization. Here, we show that modifications of the postsynaptic calcium load using the calcium chelator BAPTA or photolytic calcium uncaging result in a reversal of the expected polarity of synaptic gain change. At higher concentrations, BAPTA blocks PF-LTP. These data indicate that PF-LTD requires a higher calcium threshold amplitude than PF-LTP induction and suggest that CF activity acts as a polarity switch by providing dendritic calcium transients. Moreover, previous CF-LTD induction changes the relative PF-LTD versus -LTP induction probability. These findings suggest that bidirectional cerebellar learning is governed by a calcium threshold rule operating "inverse" to the mechanism previously described at other glutamatergic synapses (BCM rule) and that the LTD/LTP induction probability is under heterosynaptic climbing fiber control. PMID- 15541317 TI - Osmotic tension as a possible link between GABA(A) receptor activation and intracellular calcium elevation. AB - Intracellular calcium concentration rises have been reported following activation of GABA(A) receptors in neonatal preparations and attributed to activation of voltage-dependent Ca(2+) channels. However, we show that, in cerebellar interneurons, GABA(A) agonists induce a somatodendritic Ca(2+) rise that persists at least until postnatal day 20 and is not mediated by depolarization-induced Ca(2+) entry. A local Ca(2+) elevation can likewise be elicited by repetitive stimulation of presynaptic GABAergic afferent fibers. We find that, following GABA(A) receptor activation, bicarbonate-induced Cl(-) entry leads to cell depolarization, Cl(-) accumulation, and osmotic tension. We propose that this tension induces the intracellular Ca(2+) rise as part of a regulatory volume decrease reaction. This mechanism introduces an unexpected link between activation of GABA(A) receptors and intracellular Ca(2+) elevation, which could contribute to activity-driven synaptic plasticity. PMID- 15541318 TI - ApTrkl, a Trk-like receptor, mediates serotonin- dependent ERK activation and long-term facilitation in Aplysia sensory neurons. AB - The Trk family of receptor tyrosine kinases plays a role in synaptic plasticity and in behavioral memory in mammals. Here, we report the discovery of a Trk-like receptor, ApTrkl, in Aplysia. We show that it is expressed in the sensory neurons, the locus for synaptic facilitation, which is a cellular model for memory formation. Serotonin, the facilitatory neurotransmitter, activates ApTrkl, which, in turn, leads to activation of ERK. Finally, inhibiting the activation of ApTrkl with the Trk inhibitor K252a or using dsRNA to inhibit ApTrkl blocks the serotonin-mediated activation of ERK in the cell body, as well as the cell-wide long-term facilitation induced by 5-HT application to the cell body. Thus, transactivation of the receptor tyrosine kinase ApTrkl by serotonin is an essential step in the biochemical events leading to long-term facilitation in Aplysia. PMID- 15541319 TI - Decreased absolute amygdala volume in cocaine addicts. AB - The amygdala is instrumental to a set of brain processes that lead to cocaine consumption, including those that mediate reward and drug craving. This study examined the volumes of the amygdala and hippocampus in cocaine-addicted subjects and matched healthy controls and determined that the amygdala but not the hippocampus was significantly reduced in volume. The right-left amygdala asymmetry in control subjects was absent in the cocaine addicts. Topological analysis of amygdala isosurfaces (population averages) revealed that the isosurface of the cocaine-dependent group undercut the anterior and superior surfaces of the control group, implicating a difference in the corticomedial and basolateral nuclei. In cocaine addicts, amygdala volume did not correlate with any measure of cocaine use. The amygdala symmetry coefficient did correlate with baseline but not cocaine-primed craving. These findings argue for a condition that predisposes the individual to cocaine dependence by affecting the amygdala, or a primary event early in the course of cocaine use. PMID- 15541320 TI - Psychological effects of a suspicious prostate cancer screening test followed by a benign biopsy result. AB - PURPOSE: To evaluate the psychological implications of an apparently false positive screening result for prostate cancer. METHODS: The sample comprised 167 men with a benign biopsy result in response to a suspicious screening test result (biopsy group) and 233 men with a normal prostate-specific antigen (PSA) test result (control group). The men responded to a questionnaire within about 6 weeks of their biopsy or PSA results. They were asked about demographic characteristics, medical history, psychological effects, biopsy experience, and prostate cancer knowledge. RESULTS: The survey response rate was 85% (400/471). The mean (+/- SD) age of respondents was 60 +/- 9 years (range, 40 to 88 years); 88% (n = 350) were white. Forty-nine percent (81/167) of men in the biopsy group reported having thought about prostate cancer either "a lot" or "some of the time", compared with 18% (42/230) in the control group (P < 0.001). In addition, 40% (67/167) in the biopsy group reported having worried "a lot" or "some of the time" that they may develop prostate cancer, compared with 8% (18/231) in the control group (P < 0.001). CONCLUSION: Men who underwent prostate biopsy more often reported having thought and worried about prostate cancer, despite having received a benign result. This underrecognized human cost of screening should be considered in the debate about the benefits and harms of prostate cancer screening. PMID- 15541321 TI - Effects of guideline-concordant antimicrobial therapy on mortality among patients with community-acquired pneumonia. AB - PURPOSE: National practice guidelines have recommended specific initial empiric antimicrobial regimens for patients with community-acquired pneumonia. Our aim was to determine the association between the use of guideline-concordant antimicrobial therapy and 30-day mortality in patients with pneumonia. METHODS: We conducted a retrospective cohort study at two tertiary teaching hospitals. Eligible patients were admitted with a diagnosis of community-acquired pneumonia, had a chest radiograph consistent with pneumonia, and had a discharge diagnosis of pneumonia. All eligible patients were identified and a random sample was abstracted. We determined whether the use of guideline-concordant antibiotics was associated with 30-day mortality in an analysis that adjusted for potential confounders using propensity scores. RESULTS: Information was obtained on 420 patients with pneumonia. The mean (+/- SD) age was 63 +/- 16 years, 355 were men, and 82 patients were initially admitted to the intensive care unit. At 30 days after presentation, 41 patients (9.8%) had died: 21 of 97 (21.7%) in the non guideline-concordant group and 20 of 323 (6.2%) in the guideline-concordant group. Antibiotics were concordant with national guidelines in 323 patients. In the regression analysis, after adjustment for the propensity score, failure to comply with antimicrobial therapy guidelines was associated with increased 30-day mortality (odds ratio = 5.7; 95% confidence interval: 2.0 to 16.0). CONCLUSION: Receipt of antimicrobial regimens concordant with national published guidelines may reduce 30-day mortality among patients hospitalized with pneumonia. PMID- 15541322 TI - Antidepressants and risk of first-time hospitalization for myocardial infarction: a population-based case-control study. AB - PURPOSE: Several studies have found an increased risk of myocardial infarction among depressed patients. Selective serotonin reuptake inhibitors (SSRIs) appear to lack the arrhythmic adverse effects of tricyclic antidepressants, and are thought to inhibit platelet aggregation. We examined whether use of different antidepressant classes is associated with a lower risk of first-time hospitalization for myocardial infarction, as compared with nonuse. METHODS: We identified 8887 cases of first-time hospitalization for myocardial infarction and 88,862 age- and sex-matched population-based controls during 1994-2002, using data from North Jutland County, Denmark. Cases and controls were stratified according to history of cardiovascular disease. All prescriptions for antidepressants before hospitalization for myocardial infarction were identified using a prescription database. Conditional logistic regression was used to estimate odds ratios of myocardial infarction associated with antidepressant use, adjusted for possible confounding factors. RESULTS: In patients with a history of cardiovascular disease, we found indications of a lower risk of myocardial infarction among those who used SSRIs (adjusted odds ratio [OR] = 0.85; 95% confidence interval [CI]: 0.62 to 1.16), nonselective serotonin reuptake inhibitors (adjusted OR = 0.83; 95% CI: 0.50 to 1.38), and other antidepressants (adjusted OR = 0.55; 95% CI: 0.31 to 0.97). There were no such associations among persons without a history of cardiovascular disease. CONCLUSION: Antidepressant use may be associated with a decreased risk of hospitalization for myocardial infarction among persons with a history of cardiovascular disease, although it remains uncertain whether there are differences by class of antidepressant. PMID- 15541323 TI - Association of body weight with condition-specific quality of life in male veterans. AB - PURPOSE: We hypothesized that obese adults with coronary heart disease, obstructive lung disease, or depression would report greater impairments in health-related quality of life owing to their angina, dyspnea, or depressive symptoms as compared with persons with normal body weight. METHODS: We analyzed cross-sectional data from the Ambulatory Care Quality Improvement Project, a multicenter study of veterans enrolled in general internal medicine clinics. Health-related quality of life was assessed using the Medical Outcomes Study Short Form-36, the Seattle Angina Questionnaire, the Seattle Obstructive Lung Disease Questionnaire, and the Hopkins Symptom Checklist for Depression. RESULTS: Compared with patients of normal weight (body mass index: 18.5 to 24.9 kg/m2), underweight patients (body mass index <18.5 kg/m2) reported health-related quality-of-life scores that were at least 5% lower (worse) in all 15 quality-of life domains examined. Patients with class III obesity (body mass index > or =40 kg/m2) reported quality-of-life scores that were at least 5% lower than those of normal weight patients in eight domains. Scores of overweight patients (body mass index: 25 to 29.9 kg/m2) were higher (better) than those of normal weight patients in 11 domains. CONCLUSION: Body mass index was strongly associated with generic- and condition-specific health-related quality of life. Our results suggest that, when considering health-related quality-of-life outcomes among veterans, the optimal body mass index may be above the "normal" range. Further research should test the validity of the 1998 National Institutes of Health body mass index categories as predictors of health outcomes among veterans. PMID- 15541324 TI - Clinician awareness of adherence to hypertension guidelines. AB - PURPOSE: Little is known about how well clinicians are aware of their own adherence to clinical guidelines, an important indicator of quality. We compared clinicians' beliefs about their adherence to hypertension guidelines with data on their actual performance. METHODS: We surveyed 139 primary care clinicians at three Veterans Affairs medical centers, asking them to assess their own adherence to hypertension guidelines. We then extracted data from the centers' clinical databases on guideline-concordant medication use and blood pressure control for patients cared for by these providers during a 6-month period. Data were collected for patients with hypertension and diabetes, hypertension and coronary disease, or hypertension with neither of these comorbid conditions. RESULTS: Eighty-six clinicians (62%) completed the survey. Each clinician saw a median of 94 patients with hypertension (mean age, 65 years). Patients were treated with an average of 1.6 antihypertensive medications. Overall, clinicians overestimated the proportion of their patients who were prescribed guideline-concordant medications (75% perceived vs. 67% actual, P <0.001) and who had blood pressure levels <140/90 mm Hg on their last visit (68% perceived vs. 43% actual, P <0.001). Among individual clinicians, there were no significant correlations between perceived and actual guideline adherence (r = 0.18 for medications, r = 0.14 for blood pressure control; P > or =0.10 for both). Clinicians with relatively low actual guideline performance were most likely to overestimate their adherence to medication recommendations and blood pressure targets. CONCLUSION: Clinicians appear to overestimate their adherence to hypertension guidelines, particularly with regards to the proportion of their patients with controlled blood pressure. This limited awareness may represent a barrier to successful implementation of guidelines, and could be addressed through the use of provider profiles and point-of-service feedback to clinicians. PMID- 15541325 TI - Life expectancy and prognostic factors for survival in patients with polycythemia vera and essential thrombocythemia. AB - PURPOSE: To assess life expectancy and prognostic factors for survival in patients with polycythemia vera and essential thrombocythemia. METHODS: The study sample consisted of 831 consecutive patients with polycythemia vera (n = 396; 4184 person-years of follow-up) or essential thrombocythemia (n = 435; 4304 person-years of follow-up). Mortality in each group was compared with the Italian population using the standardized mortality ratio (SMR) based on life expectancy data obtained from the Italian Institute of Statistics. RESULTS: The 15-year survival was 65% in patients with polycythemia and 73% in those with thrombocythemia. By Cox regression analysis, the independent predictors of death were a history of thrombosis for polycythemia (hazard ratio [HR] = 2.2; P = 0.0002) and thrombocythemia (HR = 2; P = 0.01), and male sex (HR = 1.8; P = 0.03) for thrombocythemia. Mortality compared with the general population was 1.6-fold higher (P <0.001) in patients with polycythemia but was not increased in those with thrombocythemia (SMR = 1; P = 0.8). CONCLUSION: Life expectancy of patients with polycythemia vera (especially if younger than 50 years) was reduced compared with the general population, whereas life expectancy of patients with essential thrombocythemia was not affected significantly by the disease, reflecting the more indolent nature of the proliferation. History of thrombosis was the main predictor of death in both diseases. PMID- 15541326 TI - Long-term effectiveness of lifestyle and behavioral weight loss interventions in adults with type 2 diabetes: a meta-analysis. AB - BACKGROUND: Most persons with type 2 diabetes are overweight, and obesity worsens the metabolic and physiologic abnormalities associated with diabetes. Our objective was to assess the effectiveness of lifestyle and behavioral weight loss and weight control interventions in adults with type 2 diabetes. METHODS: Studies were obtained from searches of multiple electronic bibliographic databases, supplemented with hand searches of selected journals and consultation with experts in obesity research. Studies were included if they were published or unpublished randomized controlled trials in any language that examined weight loss or weight control strategies using one or more dietary, physical activity, or behavioral interventions, with a follow-up interval of at least 12 months. Effects were combined using a random-effects model. RESULTS: The 22 studies of weight loss interventions identified yielded a total of 4659 participants with a follow-up of 1 to 5 years. The pooled weight loss for any intervention in comparison with usual care among 585 subjects was 1.7 kg (95% confidence interval [CI]: 0.3 to 3.2 kg), or 3.1% of baseline body weight among 511 subjects. Among 126 persons who underwent a physical activity and behavioral intervention, those who also received a very low-calorie diet lost 3.0 kg (95% CI: -0.5 to 6.4 kg), or 1.6% of baseline body weight, more than persons who received a low-calorie diet. Among 53 persons who received identical dietary and behavioral interventions, those who received a more intense physical activity intervention lost 3.9 kg (95% CI: -1.9 to 9.7 kg), or 3.6% of baseline body weight, more than those who received a less intense or no physical activity intervention. Comparison groups often achieved substantial weight loss (up to 10.0 kg), minimizing between-group differences. Changes in glycated hemoglobin level generally corresponded to changes in weight and were not substantial when between group differences were examined. CONCLUSION: Weight loss strategies involving dietary, physical activity, or behavioral interventions were associated with small between-group improvements in weight. These results were minimized by weight loss in the comparison group, however, and examination of individual study arms revealed that multicomponent interventions, including very low-calorie diets or low-calorie diets, may hold promise for achieving weight loss in adults with type 2 diabetes. PMID- 15541327 TI - Effects of statins on vascular structure and function: a systematic review. AB - PURPOSE: Statins reduce cardiovascular events by more than can be explained by their effects on lipids. We conducted a systematic review of how statins affect vascular structure and function, differences among statins, and correlations between the effects of statins on vascular outcomes and either lipid levels or cardiovascular outcomes. METHODS: We primarily searched MEDLINE (1980 to March 2004) to identify all studies with at least 10 subjects that reported the effects of currently available statins on coronary artery stenosis, carotid intima-media thickness, and endothelial function (excluding studies of drug combinations and subjects with organ transplants). Meta-analyses were performed when feasible. RESULTS: Statins decrease the progression and increase the regression of coronary artery lesions and luminal narrowing. Compared with placebo, statins decrease the likelihood of coronary artery restenosis (summary risk ratio = 0.85; 95% confidence interval: 0.77 to 0.95). Statins appear to slow the progression of carotid artery intima-media thickness. Although the effect of statins on coronary endothelial function is uncertain, statins appear to improve peripheral endothelial function. There is no conclusive evidence to suggest that individual statins differ in their effects on these outcomes. Studies generally found weak or no correlation between the effects of statins on vascular outcomes and lipid levels. No study showed a correlation between vascular effect and clinical outcome. CONCLUSION: Statins slow the progression of, and may reverse, atherosclerosis. The magnitude of these effects, however, is small compared with the effects of statins on cardiovascular events. Statins also improve measures of vascular function, which may contribute to their clinical benefits. There is insufficient evidence to suggest that individual statins differ in their vascular effects. PMID- 15541328 TI - Incidence and predictors of renal disease in Chinese patients with systemic lupus erythematosus. PMID- 15541329 TI - Reducing PSAnxiety: The importance of noninvasive chronic disease management in prostate cancer detection and treatment. PMID- 15541330 TI - Community-acquired pneumonia guidelines: Peering back through the looking glass...clearly? PMID- 15541332 TI - Why not test for hypercoagulability in patients with idiopathic venous thromboembolism? PMID- 15541333 TI - A 1-week course of corticosteroids in the treatment of eosinophilic meningitis. PMID- 15541334 TI - Treatment of severe hypercholesterolemia with atorvastatin in congenital analbuminemia. PMID- 15541335 TI - Wernicke encephalopathy--an emerging trend after bariatric surgery. PMID- 15541336 TI - Advancing academic subspecialty medicine: the agenda for the future of the Association of Subspecialty Professors. PMID- 15541337 TI - The problem of protein kinase activity of small heat shock protein Hsp22 (H11 or HspB8). AB - The recently described protein denoted H11, Hsp22 or HspB8 seems to participate in regulation of proliferation, apoptosis, and cardiac hypertrophy. Mutation of Hsp22 causes distal motor neuropathy. Multitude action of Hsp22 is supposed to be due to its protein kinase and/or chaperone-like activities. There are many indirect evidences indicating that Hsp22 possesses intrinsic protein kinase activity. However, low homology to protein kinases, low extent of autophosphorylation, lack of significant protein kinase activity with commonly used substrates, and lack of information on stoichiometry, kinetics, and substrate specificity make the existence of intrinsic protein kinase activity of Hsp22 questionable. It is supposed that protein kinase activity ascribed to Hsp22 is due to contaminating protein kinases. Hsp22 is highly homologous to small heat shock proteins and effectively prevents aggregation of denatured protein both in vitro and in vivo. Therefore, it is supposed that chaperone-like activity is of great importance for Hsp22 functioning. PMID- 15541338 TI - Regulation of Rb gene expression by an MBD2-interacting zinc finger protein MIZF during myogenic differentiation. AB - The induction of Rb gene expression is a key event in the process of myogenic differentiation. We recently demonstrated that transcription of the Rb gene is repressed by MIZF, previously characterized as an MBD2-binding partner. Here we show the new roles of MIZF as a regulator of myogenic differentiation. MIZF mRNA was detected in undifferentiated C2C12 myoblasts but its expression decreased during myogenesis, correlating with an increase in Rb mRNA. To examine the function of MIZF in regulating myogenic differentiation, we transduced C2C12 myoblasts with adenoviral vectors to constitutively produce MIZF at high levels. When switched to differentiation medium, these cells showed decreased expression of Rb as well as differentiation markers such as myogenin and Troponin-T, and consequently could not differentiate into multinucleated myotubes. These results suggest that transcriptional repression of Rb by MIZF could be one of the critical determinants in myogenic differentiation. PMID- 15541339 TI - C75 activates malonyl-CoA sensitive and insensitive components of the CPT system. AB - Carnitine palmitoyltransferase I (CPT-I) and II (CPT-II) enzymes are components of the carnitine palmitoyltransferase shuttle system which allows entry of long chain fatty acids into the mitochondrial matrix for subsequent oxidation. This system is tightly regulated by malonyl-CoA levels since this metabolite is a strong reversible inhibitor of the CPT-I enzyme. There are two distinct CPT-I isotypes (CPT-Ialpha and CPT-Ibeta), that exhibit different sensitivity to malonyl-CoA inhibition. Because of its ability to inhibit fatty acid synthase, C75 is able to increase malonyl-CoA intracellular levels. Paradoxically it also activates long-chain fatty acid oxidation. To identify the exact target of C75 within the CPT system, we expressed individually the different components of the system in the yeast Pichia pastoris. We show here that C75 acts on recombinant CPT-Ialpha, but also on the other CPT-I isotype (CPT-Ibeta) and the malonyl-CoA insensitive component of the CPT system, CPT-II. PMID- 15541340 TI - Effect of human umbilical cord blood cells on glycemia and insulitis in type 1 diabetic mice. AB - Several studies have shown that transplantation of embryonic stem cells into diabetic animals either improved or normalized blood glucose levels. In this study, we examined the dose-dependent effect of early (prediabetic stage) intravenous administration of human umbilical cord blood (HUCB) mononuclear cells on blood glucose levels, survival, and insulitis in nonobese diabetic (NOD) mice with autoimmune type 1 diabetes. The results show that mice treated with HUCB cells significantly lowered their blood glucose levels and increased their lifespan, as compared with untreated mice. Also, a significant reduction in insulitis was observed in treated than in untreated mice. The mice that received the highest dosage (200 x 10(6)) of cells had greater reduction in blood glucose levels and the degree of insulitis than the mice that received lower dosage (100 150 x 10(6)) of cells. Prolonged lifespan in the former group of mice seems to be related to better control of blood glucose levels. Thus, administration of HUCB cells in the prediabetic stage without any immunosuppression improves type 1 diabetes by protecting the islets from insulitis in NOD mice. PMID- 15541341 TI - An RNA ligand inhibits hepatitis C virus NS3 protease and helicase activities. AB - The hepatitis C virus non-structural protein 3 (HCV NS3) possesses both protease and helicase activities that are essential for viral replication. In a previous study, we obtained RNA aptamers that specifically and efficiently inhibited NS3 protease activity (G9 aptamers). In order to add helicase-inhibition capability, we attached (U)14 to the 3'-terminal end of a minimized G9 aptamer, DeltaNEO-III. NEO-III-14U was shown to inhibit the NS3 protease activity more efficiently than the original aptamer and, furthermore, to efficiently inhibit the unwinding reaction by NS3 helicase. In addition, NEO-III-14U has the potential to diminish specific interactions between NS3 and the 3'-UTR of HCV-positive and -negative strands. NEO-III-14U showed effective inhibition against NS3 protease in living cells. PMID- 15541342 TI - Inhibition of hepatocyte growth factor induction in human dermal fibroblasts by interleukin-1 and its prevention by interferon-gamma. AB - Hepatocyte growth factor (HGF) is one of the vital factors for liver regeneration. HGF production is induced by the activation of protein kinase A and protein kinase C-mediated pathways, interleukin (IL)-1, tumor necrosis factor (TNF)-alpha, and epidermal growth factor (EGF) in mesenchymal cells. We here report that IL-1 and TNF-alpha, hitherto regarded as HGF inducers, potently inhibited HGF production stimulated by other HGF inducers. IL-1alpha, IL-1beta, and TNF-alpha alone had minimal stimulating effects on HGF production in human dermal fibroblasts, but they strongly inhibited production of HGF induced by cholera toxin, 8-bromo-cAMP, EGF, and phorbol 12-myristate 13-acetate (PMA). Moreover, although the high level of HGF production in MRC-5 cells was enhanced by PMA and less markedly by IL-1beta, HGF production in MRC-5 cells treated with PMA plus IL-1beta was less than that in the cells treated with PMA alone. In the presence of interferon (IFN)-gamma, however, cholera toxin- and 8-bromo-cAMP induced HGF production was not inhibited by IL-1beta. Pretreatment of cells with IL-1beta suppressed the phosphorylation of cAMP-responsive element-binding protein induced by cholera toxin but not that induced by 8-bromo-cAMP. Taken together, our results indicate that IL-1 inhibited HGF production stimulated by various inducers, including protein kinase A-activating agents, and that IFN gamma overcame this inhibition of induction of HGF production. PMID- 15541343 TI - Differential protein acetylation induced by novel histone deacetylase inhibitors. AB - Histone deacetylase (HDAC) inhibitors induce the hyperacetylation of nucleosomal histones in carcinoma cells resulting in the expression of repressed genes that cause growth arrest, terminal differentiation, and/or apoptosis. In vitro selectivity of several novel hydroxamate HDAC inhibitors including succinimide macrocyclic hydroxamates and the non-hydroxamate alpha-ketoamide inhibitors was investigated using isolated enzyme preparations and cellular assays. In vitro selectivity for the HDAC isozymes (HDAC1/2, 3, 4/3, and 6) was not observed for these HDAC inhibitors or the reference HDAC inhibitors, MS-275 and SAHA. In T24 and HCT116 cells these compounds caused the accumulation of acetylated histones H3 and H4; however, the succinimide macrocyclic hydroxamates and the alpha ketoamides did not cause the accumulation of acetylated alpha-tubulin. These data suggest "selectivity" can be observed at the cellular level with HDAC inhibitors and that the nature of the zinc-chelating moiety is an important determinant of activity against tubulin deacetylase. PMID- 15541344 TI - alpha-Trinositol inhibits FGF-stimulated growth of smooth muscle and breast cancer cells. AB - alpha-Trinositol (d-myo-inositol-1,2,6-trisphosphate), an isomer of the intracellular messenger IP(3), has been studied for its anti-inflammatory and other effects in animal experiments and in human. The mechanisms of action remain unknown. Several human pathologies are associated with uncontrolled production of fibroblast growth factors (FGFs). FGF-2 induces vascular smooth muscle cell proliferation, which contributes to restenosis after coronary balloon angioplasty. The expression of several FGFs is also increased in tumors. We studied the effects of the water- and lipid-soluble derivatives of alpha trinositol on the FGF-2- and/or FGF-8-induced proliferation of human pulmonary artery smooth muscle cells (HPASMC) and S115 mouse breast cancer cells. alpha Trinositol decreased the FGF-mediated proliferation of HPASMC and S115 cells. Membrane permeability did not seem obligatory since the lipid-soluble form of alpha-trinositol was less effective than the water-soluble derivative. These results suggest a new biological function for certain phosphoinositides in the modulation of FGF-regulated processes. PMID- 15541345 TI - IGFBP-4 degradation by pregnancy-associated plasma protein-A in MC3T3 osteoblasts. AB - Insulin-like growth factor (IGF) signaling is critical for osteoblast development and IGF binding protein (IGFBP)-4 is one of the principle IGFBPs expressed by osteoblasts. Release of bound IGF via proteolytic degradation of IGFBP-4 is likely to be critical for osteoblast development. We have investigated whether IGF-sensitive, IGFBP-4 degradation in mouse MC3T3-E1 osteoblasts is due to the metzincin pregnancy-associated plasma protein (PAPP)-A. Degradation of IGFBP-4 by PAPP-A or MC3T3-E1 conditioned medium was enhanced by IGF-II but inhibited by mutation of basic residues at or near the PAPP-A cleavage site in IGFBP-4. Furthermore, immunodepletion of PAPP-A from MC3T3-E1 conditioned medium abolished IGFBP-4 degradation. We also found that PAPP-A messenger RNA was expressed throughout differentiation of MC3T3-E1 cells. These results demonstrate for the first time that PAPP-A is the IGFBP-4 protease in MC3T3-E1 cells, a widely used model for osteoblast development, and that PAPP-A may regulate IGF release throughout osteoblast differentiation. PMID- 15541346 TI - Trimethylamine-N-oxide modulates the reductive unfolding of onconase. AB - The physiological osmolyte trimethylamine-N-oxide (TMAO) stabilizes proteins by decreasing the entropy of the unfolded state through a solvophobic effect. Our studies on the effect of TMAO on the reductive unfolding of onconase (ONC) to form its reductive intermediate, des [30-75], indicate that TMAO diminishes the reductive unfolding rate of the protein although it does not significantly affect the stability of the native protein relative to its denatured state. Since the reductive unfolding of ONC is a local event, our studies provide direct evidence for a TMAO-induced local structural change that reduces the rate of redox dependent protein unfolding. The implications of our findings for protein folding/unfolding are discussed. PMID- 15541347 TI - Poliovirus sampling by using sodium dodecyl sulfate/EDTA-pretreated chromatography paper strips. AB - To achieve the goal of poliovirus eradication, surveillance of endemic areas is a crucial step in the poliovirus eradication program. Currently, six countries still have endemic poliovirus. We have tested a novel method which uses SDS/EDTA treated chromatography paper strips to collect and transport poliovirus containing stool samples. The SDS/EDTA-treated paper strips were soaked with different dilutions of poliovirus-containing feces and stored at different temperatures. After storing the SDS/EDTA paper strips for 5 months at 37 degrees C, poliovirus RNA could be successfully amplified using RT-PCR. Infectivity of wild-type poliovirus type 1, 2, and 3 was lost upon contact with the SDS/EDTA treated strips. This easy, inexpensive, and biosafe chromatography paper strip method for the collection and transportation of poliovirus samples can be of use in poliovirus surveillance and polio vaccination programs. PMID- 15541348 TI - Stabilization, not polymerization, of microtubules inhibits the nuclear translocation of STATs in adipocytes. AB - Signal transducers and activators of transcriptions (STATs) are a family of latent transcription factors which are activated by a variety of growth factors and cytokines in many cell types. However, the mechanism by which these transcription factors translocate to the nucleus is poorly understood. The goal of this study was to determine the requirement of microfilaments and microtubules for cytokine induced STAT activation in cultured adipocytes. We used seven different actin-specific and microtubule-specific agents that are well established effectors of these cytoskeletal networks. Our results clearly demonstrate that inhibition of microfilaments or the prevention of microtubule polymerization has no effect on the ability of STATs to be tyrosine phosphorylated or to translocate to the nucleus. However, we observed that paclitaxel, a microtubule stabilizer, resulted in a significant decrease in the nuclear translocation of STATs without affecting the cytosolic tyrosine phosphorylation of these transcription factors. In summary, our results demonstrate that the dynamic instability, but not the polymerization, of microtubules contributes to nuclear translocation of STAT proteins in adipocytes. PMID- 15541349 TI - Neuroglobin, cytoglobin, and a novel, eye-specific globin from chicken. AB - Neuroglobin and cytoglobin are two recently discovered respiratory proteins of vertebrates. Here we report the first identification and expression analyses of these proteins in bird species. Neuroglobin from the domestic chicken Gallus gallus differs in approximately 30% from the mammalian proteins, but its genome structure shows the conservation of the B12.2, E11.0, and G7.0 intron positions. The chicken cytoglobin protein is shorter than the mammalian orthologs, from which it differs overall by approximately 25%, due to the absence of the C terminal exon in the gene. Comparison of chicken and mammalian gene order shows that neuroglobin and cytoglobin are located on conserved syntenic chromosomal segments. While neuroglobin is expressed in the chicken's brain and eye, cytoglobin RNA was detected in all investigated tissues. In addition, a novel globin-type has been identified that is only expressed in the chicken's eye. The gene of this eye-globin contains the typical globin introns at B12.2 and G7.0. Phylogenetic analyses suggest that this globin is most closely related to the cytoglobin lineage. Although the function of this eye-globin remains presently uncertain, it adds an additional diversity to the vertebrate globin family. PMID- 15541350 TI - Glycogen and related polysaccharides inhibit the laforin dual-specificity protein phosphatase. AB - Lafora disease, a progressive myoclonus epilepsy, is an autosomal recessive disease caused in approximately 80% of cases by mutation of the EPM2A gene, which encodes a dual specificity protein phosphatase called laforin. In addition to its phosphatase domain, laforin contains an N-terminal carbohydrate-binding domain (CBD). Mouse laforin was expressed as an N-terminally polyHis tagged protein in Escherichia coli and purified close to homogeneity. The enzyme was active towards p-nitrophenylphosphate (50-80mmol/min/mg, K(m) 4.5mM) with maximal activity at pH 4.5. Laforin binds to glycogen, as previously shown, and caused potent inhibition, half maximally at approximately 1mug/ml. Less branched glucose polymers, amylopectin and amylose, were even more potent, with half maximal inhibition at 10 and 100ng/ml, respectively. With all polysaccharides, however, inhibition was incomplete and laforin retained 20-30% of its native activity at high polysaccharide concentrations. Glucose and short oligosaccharides did not affect activity. Substitution of Trp32 in the CBD by Gly, a mutation found in a patient, caused only a 30% decrease in laforin activity but abolished binding to and inhibition by glycogen, indicating that impaired glycogen binding is sufficient to cause Lafora disease. PMID- 15541351 TI - In vitro non-natural amino acid mutagenesis using a suppressor tRNA generated by the cis-acting hepatitis delta virus ribozyme. AB - In vitro non-natural amino acid mutagenesis requires aminoacyl-charged suppressor transfer RNAs which read an internal stop codon. For the synthesis of aminoacyl tRNAs loaded with non-natural amino acids, T4 RNA ligase is used to ligate a chemically synthesised aminoacyl-dinucleotide to a truncated 74mer tRNA(-CA) lacking the two 3' end nucleotides. The 74mer tRNA(-CA) in turn is generated by run-off transcription from a linearised plasmid encoding the tRNA sequence under control of the T7 promoter. Transcripts with heterogeneous ends are commonly obtained, which interfere with subsequent reactions such as ligation or translation. Here we report an improved procedure for the generation and chromatographic purification of large amounts of homogeneous 3' end tRNA(-CA) by hepatitis delta virus ribozyme cis-cleavage and the first application of this tRNA to in vitro non-natural amino acid mutagenesis. Stop codon suppression is increased compared to conventionally synthesised suppressor tRNA; 2.5 microg of mutated protein was synthesised in a 50 microl batch reaction. PMID- 15541352 TI - Quantum dot-antibody and aptamer conjugates shift fluorescence upon binding bacteria. AB - CdSe/ZnS quantum dots (QDs) exhibited fluorescence emission blue shifts when conjugated to antibodies or DNA aptamers that are bound to bacteria. The intensity of the shifted emission peak increased with the number of bound bacteria. Curiously, the emission was consistently shifted to approximately 440 460 nm, which is distinctly different from the major component of the natural fluorescence spectrum of these QDs. This minor emission peak can grow upon conjugation to antibodies or aptamers and subsequent binding to bacterial cell surfaces. We hypothesize that the wavelength shift is due to changes in the chemical environment of the QD conjugates when they encounter the bacterial surface and may be due to physical deformation of the QD that changes the quantum confinement state. Regardless of the mechanism, these remarkable emission wavelength shifts of greater than 140 nm in some cases strongly suggest new applications for QD-receptor conjugates. PMID- 15541353 TI - Suppression of the facile latency transition of alpha(1)-antitrypsin variant M(malton) by stabilizing mutations. AB - Many genetic variants of alpha(1)-antitrypsin (alpha(1)AT) are associated with early onset emphysema and liver cirrhosis. We previously found that although the stability and inhibitory activity of the human alpha(1)AT variant M(malton) (Phe52-deleted) are comparable to those of wild-type alpha(1)AT, the M(malton) variant spontaneously undergoes a conformational change to a more stable, inactive, latent form under physiological conditions. Here, we show that insertion of an exogenous peptide having a sequence corresponding to the first strand of beta-sheet C (s1C) is facilitated in M(malton) alpha(1)AT, suggesting that the endogenous s1C and reactive center loop are easily released from beta sheet C, thus promoting latency conversion. When additional stabilizing mutations were introduced into M(malton) alpha(1)AT, they suppressed the conformational defect of this variant: the latency transition was greatly retarded, presumably by strengthening the interactions between s1C and beta-sheet C. PMID- 15541354 TI - The HIV-1 co-receptor CCR5 binds to alpha-catenin, a component of the cellular cytoskeleton. AB - The chemokine receptors CCR5 and CXCR4 belong to the family of seven transmembrane-spanning G protein-coupled receptors, which have diverse functions in host cell defense and are associated with numerous diseases. CCR5 and CXCR4 are known as co-receptors for entry of HIV-1. In this study the intracellular carboxy-terminus of CCR5, which is deleted in HIV-infected long-term non progressors, was shown to interact with the carboxy-terminus of alpha-catenin, a component of the cytoskeleton, in a yeast two-hybrid screen. This interaction was verified in mammalian cells. Furthermore, the interaction of alpha-catenin with CCR5 and CXCR4 at endogenous protein levels was demonstrated in PM1 T lymphocytes, a host cell line of HIV-1. Our results suggest that alpha-catenin links CCR5 and CXCR4 to the cytoskeleton and is involved in the organization of these receptors at the membrane, thereby possibly affecting HIV-1 infection. PMID- 15541355 TI - Induction of mouse c-src in RAW264 cells is dependent on AP-1 and NF-kappaB and important for progression to multinucleated cell formation. AB - C-src is known to play an essential role in osteoclastogenesis. We studied the regulatory mechanism as well as the significance of c-src induction in RANKL induced differentiation of mouse monocytic RAW264 cells to TRAP-positive multinucleated cells. We determined the genomic organization of the 5'-terminal region of mouse c-src. Mutational and biochemical analyses in the region 0.9 kb upstream of the transcription start site revealed that c-Fos and JNK pathways, in addition to NF-kappaB, participate in c-src induction in response to RANKL. On the other hand, when the expression of c-src was suppressed by introducing antisense src, the number of multinucleated cells formed was significantly reduced. Together, these findings show that the expression of c-src is under the control of AP-1 and NF-kappaB in the differentiation of RAW264 cells and that c src plays an essential role at the stage of progression to multinucleated cell formation. PMID- 15541356 TI - Designing effective siRNAs with off-target control. AB - Successful gene silencing by RNA interference requires a potent and specific depletion of the target mRNA. Target candidates must be chosen so that their corresponding short interfering RNAs are likely to be effective against that target and unlikely to accidentally silence other transcripts due to sequence similarity. We show that both effective and unique targets exist in mouse, fruit fly, and worm, and present a new design tool that enables users to make the trade off between efficacy and uniqueness. The tool lists all targets with partial sequence similarity to the primary target to highlight candidates for negative controls. PMID- 15541357 TI - MAP, a protein interacting with a tumor suppressor, merlin, through the run domain. AB - Merlin (or schwannomin) is a tumor suppressor encoded by the neurofibromatosis type 2 gene. Many studies have suggested that merlin is involved in the regulation of cell growth and proliferation through interactions with various cellular proteins. To better understand the function of merlin, we tried to identify the proteins that bind to merlin using the yeast two-hybrid screening. Characterization of the positive clones revealed a protein of 749 amino acids named merlin-associated protein (MAP), which showed wide tissue distribution in Northern blot analysis. Sequence analysis revealed that MAP is a potential homologue of a yeast check-point protein, BUB2, and contains TBC, SH3, and RUN domains, thereby implicating its role in the Ras-like GTPase signal pathways. MAP and merlin were directly associated in vitro and in vivo, and colocalized in NIH3T3 cells. The RUN domain of MAP and the C-terminus of merlin appeared to be responsible for their interaction. MAP decreased the AP-1-dependent promoter activity additively with merlin in NIH3T3 cells. In addition, merlin and MAP synergistically reduced the colony formation of NIH3T3 cells. These results suggest that MAP may play a cooperative role in the merlin-mediated growth suppression of cells. PMID- 15541358 TI - Heterogeneity within and between primary colorectal carcinomas and matched metastases as revealed by analysis of Ki-ras and p53 mutations. AB - Analysis of the genetic status of Ki-ras and p53 in primary colorectal carcinomas and matched colorectal liver metastasis from 30 patients reveals an overall heterogeneity both within and between the two tumoral tissues. Both genes were found mutated with a similar frequency in both tissues; however, identical mutations in primary tumor and matched metastasis were found less frequently in the case of the Ki-ras than the p53 gene. Only in three cases the same p53 and Ki ras mutations found in the primary tumor were found also in the metastasis. In several metastatic specimens the DNA bearing a mutation detected also in the primary tumor appears significantly less abundant than the wild-type DNA. These data are discussed in the light of current models of primary tumor/metastasis relationships. PMID- 15541359 TI - Cytokine-mediated induction of metallothionein in Hepa-1c1c7 cells by oleanolic acid. AB - Oleanolic acid (OA), a pentacyclic triterpene acid, has been reported to possess inducing activity of hepatic metallothionein (MT). However, the mechanism underlying its effects is unknown. This study investigated the effects of OA on the regulation of MT expression in an in vitro model. OA that was added directly to Hepa-1c1c7 cells had no effect on MT induction. However, MT and its mRNA levels increased markedly when the Hepa-1c1c7 cells were cultured with the OA treated conditioned media from the RAW 264.7 cells. Co-treating the RAW 264.7 cells with OA and pentoxifylline, a TNF-alpha synthesis inhibitor, resulted in a decrease in the effects of OA on the MT induction. In the OA-exposed RAW 264.7 cell cultures, production and mRNA levels of TNF-alpha and IL-6 were increased. However, the MT induction activity was inhibited when antibodies to TNF-alpha and/or IL-6 were added to the OA-treated conditioned media from the RAW 264.7 cells. These results suggest that the up-regulation of MT expression by OA was mediated by the TNF-alpha and IL-6 released from UA-activated macrophages. PMID- 15541360 TI - Identification and characterization of a novel gene EOLA1 stimulating ECV304 cell proliferation. AB - To study the changes in gene expression in endothelial cells stimulated by lipopolysaccharide (LPS) we performed subtraction hybridization on control human umbilical vein endothelial cells (HUVEC) versus HUVEC stimulated by LPS. A novel cDNA, named endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1), was cloned from our differentially expressed EST database of HUVEC cDNA library (GenBank Accession No. ). Computational analysis showed that EOLA1 is 1404bp long, encoding a 158aa, 17.8kDa protein, mapped to chromosome Xq27.4 with 5 exons, expressed in different human normal tissues and cancer cell lines. Using the EOLA1 cDNA as bait, we performed a yeast two-hybrid screening of a human liver cDNA library and identified metallothionein 2A (MT2A) as associated protein. Stable transfection of EOLA1 stimulates ECV304 cell proliferation. Our data suggest that the physical interaction of EOLA1 and MT2A may have an important role of cell protection in inflammation reaction. PMID- 15541361 TI - Redox sensing by Escherichia coli: effects of dithiothreitol, a redox reagent reducing disulphides, on bacterial growth. AB - Escherichia coli is able to grow with a high rate under anaerobic conditions upon decrease in redox potential (E(h)) both either in slightly alkaline (pH 7.5) or acidic (pH 5.5) medium. Upon transition of E. coli MC4100 culture to stationary growth phase a decrease in E(h) from the positive values of +120 to +160 mV to the negative ones of -380 to -550 mV, and the H(2) production are observed at various pH. A redox reagent dl-dithiothreitol (DTT) in a concentration of 3mM reduces E(h) to the negative values, and increases a latent (lag) growth phase duration, as well as delays a logarithmic growth phase independently of pH. At alkaline and acidic pH the changes in membrane potential (DeltaPsi) are observed in the presence of 3mM DTT. K(+) uptake is recovered. At pH 5.5 the H(2) production is suppressed by DTT only in a higher concentration of 10 mM. The results suggest DTT effects that are in addition to the effects of E(h). The mechanism of DTT action on bacterial growth might be intermediated through thiol group modulation of the membrane proteins, which is reflected as the generation of DeltaPsi as well as K(+) accumulation and the activity of the membrane associated enzymes. PMID- 15541362 TI - Green tea polyphenols as inhibitors of ribonuclease A. AB - Ribonucleases (RNases), which are essential for cleavage of RNA, may be cytotoxic due to undesired cleavage of RNA in the cell. The quest for small molecule inhibitors of members of the ribonuclease superfamily has become indispensable with a growing number exhibiting unusual biological properties. Thus, inhibitors of RNases may serve as potential drug candidates. Green tea catechins (GTC), particularly its major constituent (-)-epigallocatechin-3-gallate (EGCG), have reported potential against cell proliferation and angiogenesis induced by several growth factors including angiogenin, a member of the RNase superfamily. This study reports the inhibition of bovine pancreatic ribonuclease A (RNase A) by EGCG and GTC. This has been checked qualitatively by an agarose gel based assay. Enzyme kinetic studies with cytidine 2',3' cyclic monophosphate as the substrate have also been conducted. Results indicate substantial inhibitory activity of a noncompetitive nature with an inhibition constant of approximately 80 microM for EGCG and approximately 100 microM for GTC measured in gallic acid equivalents. PMID- 15541363 TI - Nuclear factor 1 regulates adipose tissue-specific expression in the mouse GLUT4 gene. AB - Previous studies demonstrated that an adipose tissue-specific element(s) (ASE) of the murine GLUT4 gene is located between -551 and -506 in the 5'-flanking sequence and that a high-fat responsive element(s) for down-regulation of the GLUT4 gene is located between bases -701 and -552. A binding site for nuclear factor 1 (NF1), that mediates insulin and cAMP-induced repression of GLUT4 in 3T3 L1 adipocytes is located between bases -700 and -688. To examine the role of NF1 in the regulation of GLUT4 gene expression in white adipose tissues (WAT) in vivo, we created two types of transgenic mice harboring mutated either 5' or 3' half-site of NF1-binding sites in GLUT4 minigene constructs. In both cases, the GLUT4 minigene was not expressed in WAT, while expression was maintained in brown adipose tissue, skeletal muscle, and heart. This was an unexpected finding, since a -551 GLUT4 minigene that did not have the NF1-binding site was expressed in WAT. We propose a model that explains the requirement for both the ASE and the NF1-binding site for expression of GLUT4 in WAT. PMID- 15541364 TI - Arachidonic acid activates tissue transglutaminase and stress fiber formation via intracellular reactive oxygen species. AB - We have investigated whether arachidonic acid could regulate tissue transglutaminase (tTGase) via intracellular reactive oxygen species (ROS) in NIH3T3 cells. tTGase was identified in NIH3T3 cells by Western blot and confocal microscopy. Arachidonic acid elevated in situ tTGase activity in dose- and time dependent manners with a maximal level at 1h, and ROS scavengers, N-(2 mercaptopropionyl)glycine and catalase, blocked the tTGase activation by arachidonic acid. The activation of tTGase by arachidonic acid was largely inhibited by transfection of tTGase siRNA. The role of intracellular ROS in the activation of in situ tTGase was supported by the activation of in situ tTGase by exogenous H(2)O(2). Arachidonic acid stimulated the formation of stress fibers in a dose- and time-dependent manner, and the ROS scavengers suppressed the arachidonic acid-induced formation of stress fibers. These results suggested that the activation of in situ tTGase and stress fiber formation by arachidonic acid was mediated by intracellular ROS in NIH3T3 cells. PMID- 15541366 TI - Identification of a novel splicing form of zebrafish p73 having a strong transcriptional activity. AB - p73 is a transcriptional activator related to tumor suppressor p53 and regulates differentiation, cell-cycle arrest, and apoptosis. Recently, zebrafish p73 (zp73alpha) was isolated and shown to be required for zebrafish embryogenesis. In this study, we isolated a novel splicing-variant of zp73 mRNA, which was generated by the use of an alternative splicing acceptor site, and designated it as zp73theta;. The zp73theta; mRNA encoded a carboxy-terminal structure distinct from that of zp73alpha. Whereas the expression level of zp73theta; mRNA was much lower than that of zp73alpha in zebrafish adult tissues, it was relatively high and fluctuated during embryogenesis. Using Saos-2 cells for a transient reporter assay, we found that zp73theta;, but not zp73alpha, had strong transcriptional activity when the experiments were performed at 34 degrees C. In addition, zp73theta; had the ability to suppress the growth of Saos-2 cells and to cause the developmental defects in zebrafish. These data indicated that zp73theta; could work as a transcriptional activator in zebrafish. PMID- 15541365 TI - mRNA levels of SREBP-1c do not coincide with the changes in adipose lipogenic gene expression. AB - Physiological differences in lipid metabolism exist according to adipose sites. To delineate at which step such gene regulation could occur, mRNA levels of various proteins involved in the overall lipogenic process were determined in subcutaneous (SC) and retroperitoneal (RP) adipose tissues. Fatty acid synthase, malic enzyme, ATP citrate lyase, insulin-sensitive glucose transporter, and glucose-6-phosphate dehydrogenase mRNA levels were coordinately reduced (by up to 50-fold) during fasting in RP and in SC relative to fed rats, and restored or overexpressed (by up to 5- to 6-fold) during refeeding. The response was most often delayed and lower in SC compared to RP. This could contribute to site specific differences. Interestingly, SREBP-1c mRNA levels were markedly decreased by fasting in SC but remained unchanged in RP. Refeeding tended to restore levels close to fed group values. We conclude that mRNA levels of SREBP-1c do not coincide with the expected changes in adipose lipogenic gene expression of fasted/refed rats. PMID- 15541367 TI - Activation and translocation of PKCdelta is necessary for VEGF-induced ERK activation through KDR in HEK293T cells. AB - VEGF-KDR/Flk-1 signal utilizes the phospholipase C-gamma-protein kinase C (PKC) Raf-MEK-ERK pathway as the major signaling pathway to induce gene expression and cPLA2 phosphorylation. However, the spatio-temporal activation of a specific PKC isoform induced by VEGF-KDR signal has not been clarified. We used HEK293T (human embryonic kidney) cells expressing transiently KDR to examine the activation mechanism of PKC. PKC specific inhibitors and human PKCdelta knock-down using siRNA method showed that PKCdelta played an important role in VEGF-KDR-induced ERK activation. Myristoylated alanine-rich C-kinase substrate (MARCKS) translocates from the plasma membrane to the cytoplasm depending upon phosphorylation by PKC. Translocation of MARCKS-GFP induced by VEGF-KDR stimulus was blocked by rottlerin, a PKCdelta specific inhibitor, or human PKCdelta siRNA. VEGF-KDR stimulation did not induce ERK phosphorylation in human PKCdelta knockdown HEK293T cells, but co-expression of rat PKCdelta-GFP recovered the ERK phosphorylation. Y311/332F mutant of rat PKCdelta-GFP which cannot be activated by tyrosine-phosphorylation but activated by DAG recovered the ERK phosphorylation, while C1B-deletion mutant of rat PKCdelta-GFP, which can be activated by tyrosine-phosphorylation but not by DAG, failed to recover the ERK phosphorylation in human PKCdelta-knockdown HEK293T cell. These results indicate that PKCdelta is involved in VEGF-KDR-induced ERK activation via C1B domain. PMID- 15541368 TI - Junctophilin type 2 is associated with caveolin-3 and is down-regulated in the hypertrophic and dilated cardiomyopathies. AB - Functional coupling between the sarcolemmal membrane and the sarcoplasmic reticulum is based on distinct structures called junctional membrane complexes (JMCs). Recently, junctophilins are found to be responsible for normal formation of JMCs. In the present study, we found that junctophilin type 2 (JP-2), a unique isoform in the heart, was localized in caveolin-rich membranes, and that the expression of JP-2 was up-regulated during normal development and down-regulated in a hypertrophic or a dilated cardiomyopathic mouse model. The expression levels of JP-2 may be associated with the development of T-tubules and impaired Ca(2+) induced Ca(2+) release in the heart. PMID- 15541369 TI - RERJ1, a jasmonic acid-responsive gene from rice, encodes a basic helix-loop helix protein. AB - Differential screening of a cDNA library constructed using poly(A)(+) RNA from suspension-cultured rice cells treated with jasmonic acid (JA) for 1/2h yielded a cDNA of a gene tentatively named RERJ1 that is upregulated in response to exogenous JA. Northern blot analysis indicated that the RERJ1 mRNA levels peaked at 1/2-1h after the addition of jasmonic acid and then decreased gradually. RERJ1 encodes a transcriptional regulator with a basic helix-loop-helix motif. The phenotypes of transgenic rice plants overexpressing sense or antisense RERJ1 mRNA demonstrated that RERJ1 is involved in the growth inhibition of rice shoots caused by JA. Other biological functions of RERJ1 are discussed from an evolutionary standpoint. PMID- 15541370 TI - Modification of soybean sucrose synthase by S-thiolation with ENOD40 peptide A. AB - The gene ENOD40 is expressed at an early stage of root nodule organogenesis and has been postulated to play a central regulatory role in the Rhizobium-legume interaction. In vitro translation of soybean ENOD40 mRNA showed that the gene encodes two peptides of 12 and 24aa residues (peptides A and B) that bind to sucrose synthase. Here we show that the small Cys-containing peptide A binds to sucrose synthase by disulfide bond formation, which may represent a novel form of posttranslational modification of this important metabolic enzyme. Assays using nanomolar concentrations of peptide A revealed that the monomeric reduced form of this peptide binds to purified sucrose synthase. Using a cysteinyl capture strategy combined with MALDI-TOF MS analysis we identified the Cys residue C264 of soybean sucrose synthase as the binding site of peptide A. Modification of sucrose synthase with ENOD40 peptide A activates sucrose cleavage activity whereas the synthesis activity of the enzyme is unaffected. The results are discussed in relation to the role of sucrose synthase in the control of sucrose utilization in nitrogen-fixing nodules. PMID- 15541371 TI - Rosuvastatin upregulates the antioxidant defense protein heme oxygenase-1. AB - Cholesterol-independent, pleiotropic actions of HMG-CoA reductase inhibitors (statins) lead to anti-inflammatory and antioxidant actions by as yet unidentified mechanisms. This study explores the role of heme oxygenase-1 (HO-1) as target and potential mediator of rosuvastatin. In cultured human endothelial cells (ECV 304), rosuvastatin increased HO-1 mRNA and protein levels in a concentration-dependent fashion. HO-1 induction by rosuvastatin remained unaffected by mevalonate and N-nitro-L-arginine-methylester, showing that isoprenoid- and NO-dependent pathways were not involved. Pretreatment of endothelial cells with rosuvastatin reduced NADPH-dependent production of oxygen radicals. The HO-1 metabolite bilirubin, when added exogenously to the cells, virtually abolished NADPH-dependent oxidative stress. Rosuvastatin-induced inhibition of free radical formation was rescued in the presence of the HO inhibitor, tin protoporphyrin-IX. Our results demonstrate that HO-1 is a target site and antioxidant mediator of rosuvastatin in endothelial cells. This novel pathway may contribute to and partially explain the pleiotropic antiatherogenic actions of rosuvastatin. PMID- 15541372 TI - Structural basis for broad substrate specificity of earthworm fibrinolytic enzyme component A. AB - Earthworm fibrinolytic enzyme component A (EFE-a) possesses an S1 pocket, which is typical for an elastase-like enzyme, but it can still hydrolyze varieties of substrates, and it exhibits wide substrate specificity. Former structure studies suggested that the four-residue insertion after Val(217) might endow EFE-a with this specificity. Based on the native crystal structure at a resolution of 2.3A, we improved the native crystal structure to 1.8A and determined its complex structure with the inhibitor Meo-Suc-Ala-Ala-Pro-Val-CMK at a resolution of 1.9A. The final structures show that: (1) EFE-a possesses multisubstrate-binding sites interacting with the substrates; (2) significant conformation adjustment takes place at two loops binding to the N-terminal of the substrates, which may enhance the interaction between the enzyme and the substrates. These characteristics make the substrate-specificity of EFE-a less dependent on the property of its S1 pocket and may endow the enzyme with the ability to hydrolyze chymotrypsin specific substrates and even trypsin-specific substrates. PMID- 15541373 TI - High affinity HERG K(+) channel blockade by the antiarrhythmic agent dronedarone: resistance to mutations of the S6 residues Y652 and F656. AB - Pharmacological inhibition of human-ether-a-go-go-related gene (HERG) K(+) channels by structurally and therapeutically diverse drugs is associated with the 'acquired' form of long QT syndrome and with potentially lethal cardiac arrhythmias. Two aromatic amino-acid residues (Y652 and F656) on the inner (S6) helices are considered to be key constituents of a high affinity drug binding site within the HERG channel pore cavity. Using wild-type (WT) and mutant HERG channels expressed in mammalian cell lines, we have investigated HERG channel current (I(HERG)) blockade at 37+/-1 degrees C by dronedarone (DRONED), a non iodinated analogue of the Class III antiarrhythmic agent amiodarone (AMIOD). Under our conditions WT I(HERG) tails, measured at -40 mV following activating pulses to +30 mV, were blocked with IC(50) values of approximately 59 and 70 nM for DRONED and AMIOD, respectively. I(HERG) inhibition by DRONED was contingent upon channel gating, with block developing rapidly on membrane depolarization, but with no preference for activated over inactivated channels. High external [K(+)] (94 mM) reduced the potency of I(HERG) inhibition by both DRONED and AMIOD. Strikingly, mutagenesis to alanine of the S6 residue F656 (F656A) failed to eliminate blockade by both DRONED and AMIOD, whilst Y652A had comparatively little effect on DRONED but some effect on AMIOD. These findings demonstrate that high affinity drug blockade of I(HERG) can occur without a strong dependence on the Y652 and F656 aromatic amino-acid residues. PMID- 15541374 TI - Differential regulation of gene expression by RNA polymerase II in response to DNA damage. AB - Cells change their gene expression profile dynamically in various conditions. By taking the advantage of ChIP, we examined the transcription profile of Saccharomyces cerevisiae genes in response to DNA damaging agents such as MMS or 4NQO. Gene expression profiles of different groups of genes roughly correlated with that revealed by Northern blot assay or microarray method. Damage-inducible genes showed increased cross-linking signals of RNA polymerase II, TFIIH, and TFIIF, meanwhile damage repressible genes decreased them, which means that gene expression is mainly regulated at the level of transcription. Interestingly, the characteristic occupancy pattern of TFIIH and polymerase with phosphorylated carboxy-terminal domain (CTD) in promoter or in coding regions was not changed by the presence of DNA damaging agents in both non-inducible and inducible genes. ChIP data showed that the extent of phosphorylation of CTD per elongating polymerase complex was still maintained. These findings suggest that overall increase in CTD phosphorylation in response to DNA damage is attributed to the global shift of gene expression profile rather than modification of specific polymerase function. PMID- 15541375 TI - ORF390 of white spot syndrome virus genome is identified as a novel anti apoptosis gene. AB - Apoptosis serves as an important defense strategy employed by host cells against viral invasion. Many viruses contain the anti-apoptotic genes to block the defense-by-death response of host cells. In this study, we tried to identify the putative anti-apoptotic genes in white spot syndrome virus (WSSV) genome. We confirmed that actinomycin D could induce apoptosis of shrimp primary cells. However, the apoptosis triggered by actinomycin D was inhibited by WSSV infection. As mutants of Autographa californica nucleopolyhedrovirus (AcMNPV), AcMNPVDelta35k/pol+ lacks a functional P35 gene undergoing apoptosis and its infection could induce Sf9 cell apoptosis. To identify the putative apoptotic suppressor gene of WSSV, overlapping cosmid clones representing the entire WSSV genome were individually cotransfected along with genome DNA of AcMNPVDeltaP35k/pol+. Using this marker rescue assay, a WSSV DNA fragment that was able to rescue AcMNPVDeltaP35k/pol+ infection in Sf9 cells was isolated. By further sequence analysis and rescue assay, the ORF390 was identified as a novel anti-apoptotic gene. The ORF displays two putative caspase9 cleavage sites LLVETDGPS, VKLEHDGSK, and a caspase3 cleavage site EEDEVDGVP. The ORF was cloned into the pIE1 vector and then the recombinant vector was transfected into Sf9 cells. The Sf9 cells did not show obvious characteristics of apoptosis when infected with AcMNPVDeltaP35k/pol+. And the transient expression of ORF390 allowed AcMNPVDeltaP35k/pol+ replication in Sf9 cells and resulted in the formation of polyhedra successfully. The results indicate that function of ORF390 in WSSV is a kind of apoptotic suppressor like P35 in AcMNPV. PMID- 15541376 TI - A role for scavenger receptor B-I in selective transfer of rhodamine-PE from liposomes to cells. AB - We investigated the potential role of scavenger receptor B-I (SR-BI) in the selective removal of liposomal markers from blood by hepatocytes. Liposomes were labeled with [(3)H]cholesteryloleyl-ether ([(3)H]COE), 1,2-di[1-(14)C]palmitoyl phosphatidylcholine ([(14)C]PC), and N-(lissamine rhodamine-B sulfonyl) phosphatidylethanolamine (N-Rh-PE). The radiolabels were eliminated at identical rates from plasma, while N-Rh-PE was cleared twice as fast. Involvement of SR-BI in the selective removal of N-Rh-PE from liposomes was studied in transfected Chinese hamster ovary cells over-expressing SR-BI. Uptake of N-Rh-PE from liposomes containing phosphatidylserine was higher than [(3)H]COE, and was further enhanced by apolipoprotein A-I, confirming involvement of SR-BI in the selective uptake of liposomal N-Rh-PE by cells. PMID- 15541377 TI - Agonist-independent localization of the NOP receptor in detergent-resistant membrane rafts. AB - A lipid rafts/detergent-resistant membrane (DRM) fraction was prepared from recombinant HEK293 cells stably expressing the human NOP receptor fused to the green fluorescent protein EGFP (hNOPr-EGFP), and probed for the presence and functionality of the fusion protein. Fluorescence detection as well as immunoblotting with an anti-GFP antibody revealed that most of the fusion protein was recovered in the DRM fraction, wherein it mediated efficiently NOP-induced stimulation of GTPgamma(35)S binding. Recovery of hNOPr-EGFP in the DRM fraction was not affected had the cells been acutely or chronically exposed to NOP prior to detergent treatment. Therefore, in HEK cells, the NOP receptor localizes constitutively to DRMs wherein it retains ability to couple with hetero-trimeric G protein. PMID- 15541378 TI - Conformational flexibility of avidin: the influence of biotin binding. AB - Ligand binding to proteins is a key process in cell biochemistry. The interaction usually induces modifications in the unfolding thermodynamic parameters of the macromolecule due to the coupling of unfolding and binding equilibria. In addition, these modifications can be attended by changes in protein structure and/or conformational flexibility induced by ligand binding. In this work, we have explored the effect of biotin binding on conformation and dynamic properties of avidin by using infrared spectroscopy including kinetics of hydrogen/deuterium exchange. Our results, along with previously thermodynamic published data, indicate a clear correlation between thermostability and protein compactness. In addition, our results also help to interpret the thermodynamic binding parameters of the exceptionally stable biotin:AVD complex. PMID- 15541379 TI - Spontaneous mutagenesis in haploid and diploid Saccharomyces cerevisiae. AB - To obtain insights into the mechanisms of spontaneous mutations in Saccharomyces cerevisiae, we have characterized the genetic alterations that inactivate either the CAN1 gene in haploid cells or heterozygously situated in diploid cells. The mutation rate in haploid cells was 9.08 x 10(-7), 100-fold lower than that in diploid cells (1.03 x 10(-4)). In haploid cells, among 69 independent CAN1 mutations, 75% were base substitutions and 22% frameshifts. The base substitutions were both transitions (33%) and transversions (42%), with G:C-->A:T and G:C-->T:A dominating. Minus frameshifts (12%) and plus frameshifts (10%) were also observed at run and non-run bases, and at A:T and G:C pairs with almost equal efficiency. An analysis of chromosome structure in diploid yeast cells indicated that allelic crossover was the predominant event followed by gene conversion and chromosome loss. We argued that genetic alterations leading to spontaneous phenotypic changes in wild-type diploid yeast cells occurred through two steps; replication-dependent alterations of bases in either allele then recombination-dependent transfer of the mutated allele to the intact one. PMID- 15541380 TI - Inactivation of the hMSH3 mismatch repair gene in bladder cancer. AB - Deficiency in the DNA mismatch repair (MMR) is frequently involved in various cancers. The hMSH3 gene is one of the human MMR genes whose role in bladder cancer is not known. We hypothesized that down-regulation of the hMSH3 gene might be involved in bladder cancer. In this study we analyzed this gene with regard to frame-shift mutation, single nucleotide polymorphism (SNP), a 9bp repeat in exon 1, loss of heterozygosity (LOH), immunohistochemistry, and methylation status in 102 bladder cancer samples. Immunohistochemistry revealed that hMSH3 expression in bladder cancer was significant decreased compared to normal epithelium (p<0.0001). An inverse correlation with pathological grade was found. The frame shift mutation in the (A) 8 tract was lacking in bladder cancer. There was no significantly difference between bladder cancer samples and healthy controls' with regard to SNP and the 9bp repeat. In bladder cancer, presence of the codon 222 polymorphism, LOH, and the 9bp repeats in exon 1 had a correlation with either pathological stage or pathological grade. Presence of the codon 1036 polymorphism had significant correlation with pathological stage and a trend to correlation with pathological grade. After 5-aza-dC treatment, MSH3 expression was significantly enhanced in TCC and UMUC bladder cancer cells when compared to untreated cells. This is the first report suggesting that genetic and epigenetic alterations in the human MSH3 gene might play a significant role in the progression of bladder tumors. PMID- 15541381 TI - Oxidative stress in the infarcted heart: role of de novo angiotensin II production. AB - This study aimed to elucidate whether angiotensin (Ang) II generated de novo at the infarct site regulates the redox state of inflammatory cells participating in cardiac repair. On days 3-28 following ligation of the rat left coronary artery, we addressed at the infarct site: (a) the appearance and cellular origin of oxidative stress by monitoring the expression (mRNA and protein) of gp91phox (a subunit of superoxide producing NADPH oxidase) and the antioxidant superoxide dismutase (SOD), together with the presence of 3-nitrotyrosine (a marker of oxidative stress); (b) the presence of components requisite to Ang peptide generation; and (c) response to treatment with losartan (Los, 10mg/kg/day). We found at the infarct site, macrophage-derived oxidative stress appears during week 1 coincident with the appearance of components requisite to AngII generation and activity in these cells. Based on observed response to AT1 receptor antagonism with Los, we would suggest de novo AngII, in an autocrine manner, participates in the induction of oxidative stress while suppressing the expression of antioxidant defenses. PMID- 15541382 TI - The murine Muc2 mucin gene is transcriptionally regulated by the zinc-finger GATA 4 transcription factor in intestinal cells. AB - MUC2, the major mucin in the intestine, is expressed early during development and shows an altered expression pattern in intestinal bowel diseases. However, the mechanisms responsible for MUC2 expression in the intestine during these events are largely unknown. Having found putative GATA binding sites in the murine Muc2 promoter and that GATA-4 is expressed in Muc2-expressing goblet cells of the mouse small intestine, we undertook to study its regulation by this transcription factor. A panel of deletion mutants made in pGL3 vector and covering 2.2kb of the promoter were used to transfect the murine CMT-93 colorectal cancer cell line. The role of GATA-4 on Muc2 gene regulation was investigated by RT-PCR and co transfections in the presence of expression vectors encoding either wild-type or mutated GATA-4 or by mutating the GATA-4 site identified within Muc2 promoter. Four GATA-4 cis-elements were identified in the promoter by EMSA and Muc2 promoter was efficiently activated when GATA-4 was overexpressed in the cells with a loss of transactivation when those sites were either mutated or a mutated form of GATA-4 was used. Altogether, these results identify Muc2, a goblet cell marker, as a new target gene of GATA-4 and point out an important role for this factor in Muc2 expression in the intestine. PMID- 15541383 TI - Fetal cells in mother rats contribute to the remodeling of liver and kidney after injury. AB - Fetal microchimerism indicates a mixture of cells of maternal and fetal origin seen in maternal tissues during and after pregnancy. Controversy exists about whether persistent fetal microchimerism is related with some autoimmune disorders occurring during and after pregnancy. In the current experiment, an animal model in which EGFP positive cells were taken as fetal-origin cells was designed to detect the fetal microchimerism in various maternal organs. Ethanol drinking and gentamicin injection were adopted to induce liver and kidney injury simultaneously. EGFP positive cells were engrafted not only in the maternal circulation and bone marrow, but also in the liver and kidney as hepatocytes and tubular cells, respectively. These results indicate that fetal cells are engrafted to maternal hematopoietic system without apparent injury and they also contribute to the repairing process of maternal liver and kidney. PMID- 15541384 TI - Wnt11 facilitates embryonic stem cell differentiation to Nkx2.5-positive cardiomyocytes. AB - Wnt signaling plays a crucial role in the control of morphogenesis in several tissues. Herein, we describe the role of Wnt11 during cardiac differentiation of embryonic stem cells. First, we examined the expression profile of Wnt11 during the course of differentiation in embryoid bodies, and then compared its expression in retinoic acid-treated embryoid bodies with that in untreated. In differentiating embryoid bodies, Wnt11 expression rose along with that of Nkx2.5 expression and continued to increase. When the embryoid bodies were treated with retinoic acid, Wnt11 expression decreased in parallel with the decreased expression of cardiac genes. Further, treatment of embryoid bodies with medium containing Wnt11 increased the expression of cardiac marker genes. Based on these results, we propose that Wnt11 plays an important role for cardiac development by embryoid bodies, and may be a key regulator of cardiac muscle cell proliferation and differentiation during heart development. PMID- 15541385 TI - Galantamine modulates nicotinic receptor and blocks Abeta-enhanced glutamate toxicity. AB - Galantamine is a plant alkaloid that is used in the treatment of Alzheimer's disease. We have studied the effects of galantamine on beta-amyloid-enhanced glutamate toxicity using primary rat cultured cortical neurons. Nicotine and galantamine alone, and in combination, protected neurons against this neurotoxicity. The protection was not blocked by alpha4beta2 nicotinic acetylcholine receptor (nAChR) antagonists, but was partially blocked by alpha7 nAChR antagonists. Galantamine induced phosphorylation of Akt, an effector of phosphatidylinositol 3-kinase (PI3K), while PI3K inhibitors blocked the protective effect and Akt phosphorylation. The antibody FK1, which selectively blocks the allosterically potentiating ligand site on nAChR, significantly reduced the galantamine-induced protection and Akt phosphorylation. Furthermore, suppression of alpha7 nAChR using an RNA interference technique reduced Akt phosphorylation induced by galantamine. Our data suggest that neuroprotection by galantamine is mediated, at least in part, by alpha7 nAChR-PI3K cascade. PMID- 15541386 TI - Crp of Streptomyces coelicolor is the third transcription factor of the large CRP FNR superfamily able to bind cAMP. AB - The chromosomal inactivation of the unique transcription factor of Streptomyces coelicolor that displays a cyclic-nucleotide-binding domain, Crp(Sco), led to a germination-defective phenotype similar to the mutant of the adenylate cyclase gene (cya) unable to produce cAMP. By means of cAMP affinity chromatography we demonstrate the specific cAMP-binding ability of Crp(Sco), which definitely demonstrate that a Cya/cAMP/Crp system is used to trigger germination in S. coelicolor. However, electromobility shift assays with the purified Crp(Sco)-cAMP complex and the CRP-like cis-acting element of its own promoter failed. Moreover, we were unable to complement an Escherichia coli crp mutant in trans with Crp(Sco). The fact that Vfr from Pseudomonas aeruginosa and GlxR from Corynebacterium glutamicum could complement such an E. coli mutant suggests that the way Crp(Sco) interacts with DNA should mechanistically differ from its most similar members. This hypothesis was further supported by homology modelling of Crp(Sco) that confirmed an unusual organisation of the DNA-binding domain compared to the situation observed in Crp(Eco). PMID- 15541387 TI - Multiple POU-binding motifs, recognized by tissue-specific nuclear factors, are important for Dll1 gene expression in neural stem cells. AB - We cloned the 5'-flanking region of the mouse homolog of the Delta gene (Dll1) and demonstrated that the sequence between nucleotide position -514 and -484 in the 5'-flanking region of Dll1 played a critical role in the regulation of its tissue-specific expression in neural stem cells (NSCs). Further, we showed that multiple POU-binding motifs, located within this short sequence of 30bp, were essential for transcriptional activation of Dll1 and also that multiple tissue specific nuclear factors recognized these POU-binding motifs in various combinations through differentiation of NSCs. Thus, POU-binding factors may play an important role in Dll1 expression in developing NSCs. PMID- 15541388 TI - Characterization of a MAPKK-like protein kinase TOPK. AB - A MAPKK-like protein kinase TOPK expresses in a wide range of proliferating cells and tissues such as cancer cells and testis. However, details of this kinase are still uncovered. We investigated the intracellular distribution of TOPK and its association with cdk1/cyclin B and microtubules. In interphase cells, TOPK expresses in cytosol and nucleus without any significant association with microtubule networks. During mitosis, TOPK-Thr-9 was phosphorylated by cdk1/cyclin B and TOPK significantly associates with mitotic spindles. When TOPK expression was suppressed, formation of spindle midzone was thinned and dimmed and cytokinesis was disturbed. We propose that TOPK plays a role in the formation of spindle midzone and in cytokinesis. PMID- 15541389 TI - Prion protein fate governed by metal binding. AB - The conversion of the normal cellular prion protein to an abnormal isoform is considered to be causal to the prion diseases or transmissible spongiform encephalopathies. The prion protein is a copper binding protein but under some conditions may bind other metals. In particular, the binding of manganese has been suggested to convert the prion protein (PrP) to a protease resistant isoform. Therefore, the differences in the way the protein binds copper and manganese might be revealing in terms of the mechanism of conversion of the protein or its normal cellular activity. We report the use of near-infrared spectroscopy for studies on aqueous solutions of prion protein binding Cu or Mn. These alloforms of the protein were analyzed by spectral data acquisition and multivariate analysis. Our results indicate that PrP binds both Mn and Cu differently. Analyses of Cu binding suggest that the PrP-Cu complex protected Cu from the water increasing protein stability. PrP-Mn does not protect Mn from water interactions. A real-time study of the protein alloforms showed that PrP-Cu remains stable in solution, but that PrP-Mn underwent highly different changes that led to fibril formation. PMID- 15541390 TI - Semicarbazide-sensitive amine oxidase in transgenic mice with diabetes. AB - Semicarbazide-sensitive amine oxidase (SSAO) activity in plasma is increased in diabetes, and in particular, in diabetic patients with vascular complications. It has been speculated that SSAO is involved in the development of such complications due to the production of cytotoxic compounds. In this work, we have induced diabetes in a previously described mouse-model, overexpressing SSAO in smooth muscle cells. SSAO activity was estimated as well as expression of the endogenous mouse gene and human transgene using real-time PCR. Diabetes induced an increase in SSAO activity in serum, kidney, and adipose tissue of transgenic animals. An inverse correlation between SSAO activity and mouse SSAO mRNA levels was observed in transgenic animals with diabetes. These results further support the suggestion of a negative feedback control of the SSAO gene expression. The increased SSAO activity in diabetes is most likely dependent on post transcriptional modifications or activation of existing inactive enzyme molecules. PMID- 15541391 TI - Repression of transcription at the human T-cell receptor Vbeta2.2 segment is mediated by a MAX/MAD/mSin3 complex acting as a scaffold for HDAC activity. AB - The identification of protein components in complex networks of co-regulators responsible for the modulation of proliferation versus differentiation modes of cell growth is a major problem. We use a combination of surface enhanced laser desorption/ionization mass spectrometry, surface plasmon resonance coupled to electrospray mass spectrometry, and immunoelectromobility shift assays to identify members of the MAX/MAD family binding to a specific DNA silencer fragment involved in the regulation of transcription for the human T-cell receptor Vbeta2.2 segment. We also identify the cofactors mSin3 and N-CoR known to interact with histone deacetylases. Inhibition of deacetylase activity in Jurkat cells prevented transcription inhibitor complex formation at the Vbeta2.2 segment, suggesting that this is either directly or indirectly dependent on the presence of HDACs. PMID- 15541392 TI - Acid excreting mutants of yeast Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae mutants acidifying glucose medium containing bromocresol purple were shown to excrete protons when placed in unbuffered water in the absence of any external carbon source. The mutants belong to 16 different complementation groups. Most of them do not grow on glycerol and the excreted protons are associated to particular sets of organic anions such as citrate, aconitate, succinate, fumarate or malate. These novel types of respiratory mutations seem to be located in genes operating in the Krebs or glyoxylate cycle. PMID- 15541393 TI - Unusual distribution of DNA methylation within the hTERT CpG island in tissues and cell lines. AB - The promoter region of the gene encoding the human telomerase reverse transcriptase (hTERT) is located in a CpG island and was shown to be regulated, at least in part, by DNA methylation. However, the observed correlation between hTERT methylation and gene expression was opposite to the general model of regulation by DNA methylation. We established a detailed mapping of methylcytosines at the CpG island (-1539 to +1732) surrounding the hTERT promoter in tissues and cell lines. In telomerase-positive samples, a methylation of all the CpG sites was observed for the hTERT promoter region (-500 to +1), whereas the exonic part (+1 to +450) revealed an unstable methylation pattern. Incomplete methylation of the proximal exon region could be necessary for, at least, a low level of hTERT transcription. In conclusion, hypermethylation of the CpG island plays a complex but essential role in the expression of hTERT in telomerase positive cells. PMID- 15541394 TI - Butyrate regulation of glycosylation-related gene expression: evidence for galectin-1 upregulation in human intestinal epithelial goblet cells. AB - Glycosylation of mucins produced by human intestinal goblet cells plays a crucial role in their functions: mucus gel physico-chemical protective properties, host bacteria interactions, cell-cell adhesion, cell migration, and cell signaling. Colonic mucin glycosylation can be modified by luminal metabolites of fiber fermentation like butyrate. Our aim was to assess the effect of butyrate on the expression of a large panel of glycosylation-related genes in human intestinal epithelial goblet cells HT29-Cl.16E. We found that only a very scarce group of genes: 9 out of 252 were evidenced by microarray screening, and only three had their modulation significantly confirmed by real time PCR quantification. The most striking effect of butyrate was its 8- to 18-fold increase of galectin-1 gene expression, which was confirmed at the protein level, specifically with a central and apical intracellular localization. Significant butyrate effects will be discussed in regard to their possible link with mucins expressed by HT29 Cl.16E cells. PMID- 15541395 TI - FGF-2 potently induces both proliferation and DSP expression in collagen type I gel cultures of adult incisor immature pulp cells. AB - We investigated the effects of both cytokines and extracellular matrices on the proliferation and differentiation of immature adult rat incisor dental pulp cells. These immature cells, which have a high-proliferative potency in vitro and do not express mRNAs for dentin non-collagenous proteins such as dentin sialoprotein (DSP), bone sialoprotein (BSP), and osteocalcin, exist in the root regions of adult rat incisors. Fibroblast growth factor-2 (FGF-2) stimulated the proliferation of these immature cells and the subsequent production of mineralized calcium was induced by beta-glycerophosphate treatment. Additionally, FGF-2 dramatically induced the expression of DSP and BSP mRNAs, but only in collagen type I gel cultures, whereas neither plate-coated collagen type I nor fibronectin, laminin or collagen type IV cultures could produce this effect and generate sufficient physiological levels of these transcripts. Although bone morphogenetic protein-4 could not induce the proliferation of immature dental pulp cells nor upregulate DSP mRNA expression, it had a synergistic effect upon DSP transcript levels in conjunction with FGF-2. These results suggest that both the presence of FGF-2 and the three-dimensional formation of immature dental pulp cells in collagen type I gel cultures are essential for both DSP expression and odontoblast differentiation. These observations provide valuable information concerning the study of the commitment and differentiation of odontoblast lineages, and also provide a basis for the rational design of cytokine and extracellular matrix based compounds for regenerative therapies in new dental treatments. PMID- 15541396 TI - Binding of arachidonic acid to myeloid-related proteins (S100A8/A9) enhances phagocytic NADPH oxidase activation. AB - Activation of the O(2)(-) generating NADPH oxidase of phagocytes results from the assembly of the membrane-bound flavocytochrome b(558) with cytosolic proteins, p67(phox), p47(phox), and Rac. However, it has been recently reported that the arachidonic acid- and calcium-binding heterodimer S100A8/A9, abundant in neutrophil cytosol, influences the activation process. In a semi-recombinant system comprising neutrophil membranes, recombinant proteins, p67(phox), p47(phox), GTPgamma S-loaded Rac2, and arachidonic acid (AA), both the rate and the extent of the oxidase activation were increased by S100A8/A9, provided it was preloaded with AA. Binding of [(14)C]AA to S100A8/A9 was potentiated by recombinant cytosolic phox proteins and GTPgammaS, suggesting the formation of a complex, comprising oxidase activating proteins and S100A8/A9, with a greater affinity for AA. The rate constant of oxidase activation was not increased by AA loaded S100A8/A9, whereas the maximal oxidase activity elicited was twice as high. AA-loaded S100A8/A9 increases oxidase activation probably by decreasing the deactivation rate. PMID- 15541397 TI - Temperature-dependent modulation of farnesyl diphosphate/geranylgeranyl diphosphate synthase from hyperthermophilic archaea. AB - Enzyme characteristics of trans-prenyl diphosphate synthase (Tk-IdsA) from Thermococcus kodakaraensis, which catalyzes the consecutive trans-condensation of isopentenyl diphosphate (C(5)) units with allylic diphosphate, were examined. Product analysis revealed that Tk-IdsA is a bifunctional enzyme, farnesyl diphosphate (FPP, C(15))/geranylgeranyl diphosphate (GGPP, C(20)) synthase, and mainly yields both C(15) and C(20). The FPP/GGPP product ratio increases with the rise of the reaction temperature. The kinetic parameters obtained at 70 and 90 degrees C demonstrated that the rise of the temperature elevates the k(0) value for the C(10) allylic substrate to more than those for the C(5) and C(15) allylic substrates. These data suggest that Tk-IdsA contributes to adjust the membrane composition to the cell growth temperature by modulating its substrate and product specificities. Mutation study indicated that the aromatic side chain of Tyr-81 acts as a steric hindrance to terminate the chain elongation and defines the final product length. PMID- 15541398 TI - Non-induced leukocyte extract reduces HIV replication and TNF secretion. AB - According to UNAIDS, the global HIV/AIDS epidemic increased to 40 million the number of people living with the virus around the world. Dialyzable leukocyte extract obtained by our group is a low molecular weight dialyzable material from peripheral human leukocytes previously in vitro induced with Sendai virus (DLE ind), and more recently, from non-induced leukocytes (DLE n/i). Previous results have shown the ability of DLE-ind to inhibit HIV in vitro replication in MT4 cell; to reduce TNFalpha secretion, and to delay in vivo progression to AIDS in early stage of HIV infection. In this work we present evidences that DLE n/i also inhibits HIV in vitro replication and reduces TNFalpha secretion in human whole blood like DLE obtained from induced leukocytes. Taking together these results show that both properties of DLE, HIV in vitro inhibition and TNF production modulation, are not dependent on in vitro Sendai virus induction of leukocytes. PMID- 15541399 TI - Role of weak interactions in thermal stability of proteins. AB - A database analysis was done to study the role of weak interactions such as CHcdots, three dots, centeredO, CHcdots, three dots, centeredPI(m) and NHcdots, three dots, centeredPI(m) in the thermal stability of proteins. The CHcdots, three dots, centeredO and CHcdots, three dots, centeredPI(m) interactions are more in the case of thermophilic proteins as compared to mesophiles. Amino acid analysis showed that hydrophobic amino acids like Val and Ile, and Cys contribute more to CHcdots, three dots, centeredO hydrogen bonds where as Pro and Gly contribute more to CHcdots, three dots, centeredPI(m) interactions. Though NHcdots, three dots, centeredPI(m) interactions are dominated by Lys and Arg in thermophiles and mesophiles, the Arg contribution is significantly higher in thermophiles. Interestingly, Glycine is a predominant contributor to all the weak interactions. The number of aromatic amino acids in the thermophiles is more and hence a large number of aromatic clusters were observed in this class. Thus, a cumulative effect of weak interactions seems to be important in thermal stability of proteins. The study also shows that introduction of Gly, Arg, Phe, Pro, and Tyr may enhance the thermal stability. PMID- 15541400 TI - Immunochemical detection of 3-deoxyglucosone in serum. AB - 3-Deoxyglucosone (3-DG) is a metabolite of glucose that is thought to lead to the production of advanced glycation end products in diabetes. The previous assay for 3-DG in serum was based on a multi-step protocol, including derivatization, extraction, HPLC separation, and detection. In the current studies, we established a monoclonal antibody that recognizes the 3-DG-derivative, which is generated by the reaction of 3-DG and a 2,3-diamino-benzene derivative. Attachment of a biotin moiety to the 2,3-diamino-benzene ring via a linker allowed development of a highly sensitive chemiluminescent enzyme immunoassay for 3-DG equivalents. Unlike the previous assay, this method does not require extraction of 3-DG derivatives from serum. Treatment of 3-DG in serum with the DAB-link-biotin produced a quinoxaline derivative, which was specifically recognized by the monoclonal antibody. Using this assay, we found that serum 3-DG was higher in streptozotocin-induced diabetic rats than in normal control rats (25+/-5.6 vs. 9.8+/-1.1 microg/L). This simple assay may allow the monitoring of conditions leading to the accumulation of advanced glycation end products and evaluation of the risk of complications in diabetic patients. PMID- 15541401 TI - High stability of the hinge region in the membrane-active peptide helix of zervamicin: paramagnetic relaxation enhancement studies. AB - Zervamicin IIB is a 16 amino acid peptaibol that forms voltage dependent ion channels with multilevel conductance states in planar lipid bilayers and vesicular systems. Stability of the hinge region and intermolecular interactions were investigated in the N- and C-terminally spin-labelled peptide analogues. Intermolecular and intramolecular paramagnetic enhancement indicates that zervamicin behaves as a rigid helical rod in methanol solution. There are no high amplitude hinge-bending motions, and the peptaibol is monomeric up to concentration 1.5 mM. Stability of the hinge region illustrates the helix stabilising propensity of the Pro residue in membrane mimic environments and implies absence of significant conformational rearrangement due to voltage peptaibol activation. PMID- 15541402 TI - Dominant negative insulin-like growth factor-1 receptor inhibits neointimal formation through suppression of vascular smooth muscle cell migration and proliferation, and induction of apoptosis. AB - Blocking of the IGF-1 signaling pathway targeting the IGF-1 receptor (IGF-1R) provides a potential treatment strategy for restenosis. In this study, we have examined the effects of a dominant negative IGF-1R (IGF-1Rt) on primary rat VSMCs in vitro and on injured rat carotid artery in vivo. Ad/IGF-1Rt infection inhibited VSMC migration and proliferation, and it also induced apoptosis by inhibiting phosphorylation of Akt and phosphorylation of ERK1/2. Consistent with the anti-proliferative and apoptotic effects in vitro, the Ad/IGF-1Rt infection markedly reduced neointimal formation in carotid injury model. Ad/IGF-1Rt treated carotid arteries exhibited a suppressed proliferation index, PCNA expression, and also were stained positive for TUNEL assay. These results indicate that a dominant negative IGF-1R has the potential to reduce neointimal formation of injured rats' carotid arteries. The delivery of dominant negative IGF-1R by adenoviral or other vectors may provide a useful strategy for inhibiting restenosis after angioplasty. PMID- 15541403 TI - In the new year, a new beginning to end HIV/AIDS. PMID- 15541404 TI - Progestogen-only contraception in women at high risk of venous thromboembolism. AB - The objective of the study was to evaluate the venous impact of a progestogen only contraception on women at high risk of venous thromboembolism (VTE). In this retrospective cohort study, 204 consecutive women at high risk of VTE were recruited between January 1992 and June 1997 and were prospectively followed. Women using chlormadinone acetate (CMA) at antigonadotropic doses (n=102) were matched by age and date of referral and history of venous thrombosis with women who had no hormonal contraception (n=102). During follow-up (mean of 33 months), nine episodes of VTE were observed: three in women receiving CMA and six in nontreated women. Using the Cox model to adjust for confounding variables such as age, thrombophilia and body mass index, the relative risk of VTE associated with the use of CMA was not significant [relative risk: 0.8 (0.2-3.9)]. These reassuring results need to be confirmed in other prospective studies. PMID- 15541405 TI - Pituitary-ovarian function following the standard levonorgestrel emergency contraceptive dose or a single 0.75-mg dose given on the days preceding ovulation. AB - We assessed to what extent the standard dose of levonorgestrel (LNG), used for emergency contraception, or a single dose (half dose), given in the follicular phase, affects the ovulatory process during the ensuing 5-day period. Fifty-eight women were divided into three groups according to timing of treatment. Each woman contributed with three treatment cycles separated by resting cycles. All received placebo in one cycle, and standard or single dose in two other cycles, in a randomized order. The diameter of the dominant follicle determined the time of treatment. Each woman had the same diameter assigned for all her treatments. Diameters were grouped into 33 categories: 12-14, 15-17 or 18-20 mm. Follicular rupture failed to occur during the 5-day period in 44%, 50% and 36% of cycles with the standard, half dose and placebo, respectively. Ovulatory dysfunction, characterized by follicular rupture associated with absent, blunted or mistimed gonadotropin surge, occurred in 35%, 36% and 5% of standard, single dose or placebo cycles, respectively. In conclusion, LNG can disrupt the ovulatory process in 93% of cycles treated when the diameter of the dominant follicle is between 12 and 17 mm. It is highly probable that this mode of action fully accounts for the contraceptive efficacy as well as the failure rate of this method. The present data suggest that half the dose may be as effective as the standard dose. PMID- 15541406 TI - Effectiveness and acceptability of vitamin E and low-dose aspirin, alone or in combination, on Norplant-induced prolonged bleeding. AB - A study (ISRCTN 77665712) was undertaken to test the effectiveness and the acceptability of vitamin E and low-dose aspirin, alone or in combination, as treatment for prolonged vaginal bleeding induced by Norplant. A total of 486 Norplant users who were requesting treatment for bleeding lasting longer than 7 days were enrolled in five centers: Beijing, China; Jakarta, Indonesia; Santiago, Chile; Santo Domingo, Dominican Republic; and Tunis, Tunisia. They were randomized to one of four different 10-day oral treatments: 200 mg vitamin E daily, 80 mg aspirin daily, both or a placebo. Treatment packs were designed to ensure blinding of both the subjects and the clinical staff. Neither vitamin E nor low-dose aspirin nor their combination was found to have any effect on reducing the length of the bleeding episode for which treatment was taken or on the vaginal bleeding patterns these women experienced during the year of follow up. PMID- 15541407 TI - Comparing vaginal and buccal misoprostol when used after methotrexate for early abortion. AB - OBJECTIVE: The primary objective of this study was to determine if buccal misoprostol was as effective as vaginal misoprostol in medical abortions. The secondary objectives were to compare side effects and acceptability. METHODS: This was a randomized controlled trial of 600 microg misoprostol by the buccal or vaginal routes used 3 to 6 days after 50 mg/m2 methotrexate. The participants were women presenting for abortion at 49 days or less gestation. The outcome measures were the number of women who had aborted by Day 8, side effects and acceptability. RESULTS: Day 8 completion rate was 53.5% for the buccal route and 67.5% for the vaginal route (p = 0.012). Side effects were similar in the two groups except that there was more burning with the buccal route. Overall acceptability and route acceptability were similar in the two groups. CONCLUSIONS: When used after methotrexate for early abortion, the vaginal route for misoprostol is more effective and preferred to the buccal route. PMID- 15541408 TI - Examination of changes caused by tubal sterilization in ovarian hormone secretion and uterine and ovarian artery blood flow rates. AB - OBJECTIVES: To examine the changes caused by tubal sterilization (TS) in ovarian hormone secretion and uterine and ovarian circulation. DESIGN: Tubal sterilization was performed by minilaparotomy and laparoscopy methods in 36 women. Blood samples were taken for hormonal tests on Preoperative Day 3 (D3) of the menstrual cycle, on Postoperative Days 13-15 (periovulatory period) of the same cycle and on D3 in the 1st and 6th months post-TS. Uterine and ovarian artery blood flow rates of the women were measured on the same days as hormonal tests by transvaginal color Doppler ultrasonography (TVCDUSG). The control group was composed of 15 volunteers in the same age group who preferred the barrier method and who had the same TVCDUSG and hormonal analyses in the same periods. RESULTS: There was a decrease in the uterine and ovarian artery pulsatility index (PI) measurements and an increase in serum luteinizing hormone (LH) and estradiol (E2) values during the periovulatory period as compared with preoperative and postoperative menstrual measurements in all groups. There was no difference between baseline uterine and ovarian artery PI and serum follicle-stimulating hormone, LH and E2 values and those measured on D3 of the menstrual cycle in the 1st and 6th months post-TS. CONCLUSIONS: The 6-month postoperative follow-up of groups that had undergone different TS methods showed no difference in uterine or ovarian artery blood flow rates or ovarian hormone secretion in comparison with baseline values. PMID- 15541409 TI - Impact of common contraceptive methods on quality of life and sexual function in Hong Kong Chinese women. AB - The combined oral contraceptive (COC) pills, injectables, intrauterine contraceptive device (IUCD) and female sterilization are the most common contraceptive methods used by women. Women's choice, compliance and satisfaction with specific contraceptive methods are influenced by any impact of the method on their quality of life and sexual function. Anxiety regarding possible adverse effects of the contraceptive methods on their quality of life and sexual function is one of the common concerns. The aim of this prospective observational study was to determine the impact of the abovementioned contraceptive methods on the quality of life and sexual function of the users. A sample of 361 Hong Kong Chinese women who were first-time users of the following contraceptive methods completed the study: COC pills (n=87), injectables (n=67), IUCD (n=96) and female sterilization (n=111). Quality of life and sexual function of the subjects were assessed before and 3-4 months after use of the method by a standardized questionnaire. The questions were adopted from the validated Chinese versions of the World Health Organization Quality of Life (WHOQOL) questionnaire and the Derogatis Sexual Functioning Inventory (DSFI). In the female sterilization group, we found a significantly higher score for sexual satisfaction (p=.004) and sexual drive (p=.003) 3-4 months after sterilization, as well as an improved WHOQOL social domain score (p=.009). However, the other DSFI subscale scores and WHOQOL domain scores were not significantly different (p>.05). No significant difference was demonstrated in all the WHOQOL domain scores and DSFI subscale scores after use of COC pills, injectables and IUCD (p>.05). We conclude that the COC pills, injectables, IUCD and female sterilization all do not have significant adverse impact on quality of life and sexual function. After female sterilization, there is a significant improvement in sexual satisfaction and sexual drive. PMID- 15541410 TI - Acceptability of levonorgestrel subdermal implants versus tubal ligation for long term contraception in a rural population of Zimbabwe. AB - This prospective cohort study of 257 rural Zimbabwean women was designed to compare patient satisfaction with levonorgestrel subdermal implants (Norplant and reg;) versus tubal ligation (TL) as a method of long-term contraception. Women were equally satisfied with both methods. At 1 year, 96% of Norplant users and 97% of the TL group reported being "satisfied" or "very satisfied," and 98% of Norplant users, and 99% of the TL cohort would recommend their chosen method to a friend or relative. Rating scores at both 6 months and 1 year were high and equal, with women in each cohort rating their method, on average, as 9 out of 10. Norplant users were significantly more likely to rate birth spacing as a reason for satisfaction with their method, whereas women in the TL cohort were more likely to select multiparity. The 1-year continuation rate with Norplant was 90%, with 4% discontinuing in order to have more children. Selecting the most appropriate method depends primarily on the potential desire of the woman to have more children. PMID- 15541411 TI - Expanding medical abortion in Tunisia: women's experiences from a multi-site expansion study. AB - From November 2000 to July 2001, 321 consenting women were enrolled at four sites across the country in an effort to demonstrate that mifepristone medical abortion could safely be used by providers throughout Tunisia. Women who met the study's inclusion criteria were given 200 mg oral mifepristone and offered the choice of taking 400 microg oral misoprostol 2 days later either at home or at the clinic. At follow-up, women were examined to determine completed abortion status and surveyed to gauge their satisfaction with the method. Ninety-six percent of women had a successful abortion using this method. Women expressed a strong preference for home use of misoprostol, indicating that it is more confidential (34%), easier (28%) and requires fewer clinic visits (28%). The high rate of success, demonstrated safety and acceptability of the method in new facilities and with new providers suggests that medical abortion can be safely expanded to new settings with reasonable levels of training and supervision. PMID- 15541412 TI - Postcoital testing after the use of a bio-adhesive acid buffering gel (ACIDFORM) and a 2% nonoxynol-9 product. AB - ACIDFORM is a gel already shown to be safe in vaginal use during six consecutive days through colposcopic observation. A randomized, blinded, crossover study was carried out to compare the spermicidal effect of ACIDFORM to that of a commercial 2% nonoxynol-9 (N-9) product. Twenty sexually active, sterilized women, aged 19 45 years, with regular cycles, normal gynecological exam, no previous sexually transmitted infection (STI) and stable partner willing to undergo four postcoital tests (PCTs) at midcycle during four different menstrual cycles, were enrolled in the study. Known allergy to N-9 or ACIDFORM, oligospermic partner, chronic diseases, use of hormones, previous treatment for STI or presence of STI at screening tests were criteria for exclusion. ACIDFORM or N-9 product administered 0-30 min precoitus or ACIDFORM given 8-10 h precoitus significantly reduced the mean number of progressively motile sperm compared to control cycles (0.19, 0.07, 0.75 vs. 17.94, respectively, p<.05, Wilcoxon test). All treated cycles had five or fewer progressively motile sperm in midcycle cervical mucus, confirming in vivo the spermicidal effect of ACIDFORM previously observed in vitro and in animal models. PMID- 15541413 TI - In vivo distribution of a vaginal gel: MRI evaluation of the effects of gel volume, time and simulated intercourse. AB - A microbicide is designed to coat the vaginal epithelium and prevent transmission of HIV. Complete coverage is desired for optimal protection. In vivo factors affecting coverage have not yet been studied. This randomized crossover trial evaluates the effect of gel volume and patient activity upon vaginal epithelial coating. Gynol II gel was mixed with a magnetic resonance imaging (MRI) contrast agent. Ten women self-inserted, on separate visits, 3 or 5 mL of gel and underwent serial MRI scanning both before and after simulated intercourse. Gel spread was dependent upon time and volume. There was modest spread during the first hour and greater spread 6 h after insertion. Five milliliters of gel resulted in statistically significantly greater coverage immediately following insertion, within the first 30 min and at 6 h after insertion. Simulated intercourse greatly enhances gel spread. After simulated intercourse, the distribution of the gel at each volume was similar. Less leakage of gel was reported with the smaller volume. PMID- 15541415 TI - Randomized trial to compare perioperative outcomes of Filshie clip vs. Pomeroy technique for postpartum and intraoperative cesarean tubal sterilization: a pilot study. PMID- 15541416 TI - Proliferation-stimulating effects of icaritin and desmethylicaritin in MCF-7 cells. AB - Icariin, icaritin and desmethylicaritin are constituents of Epimedium with a similar structure to genistein and daidzein. Using the modified MCF-7 cell proliferation assay (E-SCREEN assessment system), these compounds were tested for their estrogen-like activities. Icaritin and desmethylicaritin, but not icariin, strongly stimulated the proliferation of MCF-7/BUS cells. Cell cycle analysis revealed that the proliferation stimulatory effect was associated with a marked increase in the number of MCF-7/BUS cells in S phase and a significant increase in the G2/M population, with effects similar to those of estradiol. These actions were dose dependent (range from 1 nM to 10 microM) and could be significantly inhibited by the specific estrogen receptor antagonist ICI 182,780 [7 alpha [9(4,4,5,5,5-pentafluoropentyl)sulfinyl]nonyl)-estra-1,3,5(10)-triene-3,17beta diol)]. The estrogen receptor-regulated progesterone receptor and PS2 mRNA levels were increased by treatment with icaritin or desmethylicaritin within 24 h and the effects were also reversed by ICI 182,780. It was concluded that icaritin and desmethylicaritin are novel phytoestrogens and that the estrogenic effects of icaritin and desmethylicaritin are mediated by the estrogen receptor. PMID- 15541417 TI - Pharmacological and functional characterization of bradykinin B2 receptor in human prostate. AB - The objective of this study was to pharmacologically characterize bradykinin receptors, a component of the kallikrein-kinin system, in normal human prostate cells. In primary cultured human prostate stromal cells, bradykinin, but not [des Arg9]bradykinin or [des-Arg10]kallidin, produced calcium mobilization or inositol phosphates accumulation with potencies (pEC50) of 8.8+/-0.2 and 8.2+/-0.2, respectively. This was consistent with abundance of bradykinin B2 mRNA over bradykinin B1 mRNA in prostate stromal cells. Although the prostate epithelial cells (prostate epithelium, BPH-1, and PC-3) expressed mRNA for bradykinin B2 receptors (albeit in lesser amounts than stromal cells), bradykinin was not functionally efficacious in the epithelial cells. Increasing concentrations of D arginyl-L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thienyl)-L-alanyl L-seryl-D-1,2,3,4-tetrahhydro-3-isoquinolinecarbonyl-L-(2alpha,3beta,7alphabeta) octahydro-1H-indole-2-carbonyl-L-arginine (HOE-140), a bradykinin B2-selective peptide antagonist, attenuated bradykinin concentration-response curves in human prostate stromal cells with apparent estimate of affinity similar to that for the human bradykinin B2 receptor. Bradykinin (10 nM) caused proliferation of prostate stromal cells and phosphorylated extracellular signal-regulated kinases (ERK-1 and ERK-2) that were blocked by HOE-140 (1 microM). This study demonstrated that, in primary cultures of normal human prostate stromal cells, bradykinin activates bradykinin B2 receptors that may play a significant role in proliferation via activation of ERK-1/2 pathways. PMID- 15541418 TI - Effect of nicotine on 3-nitropropionic acid-induced oxidative stress in synaptosomes. AB - In this paper, the effect of nicotine on the oxidative changes produced by 3 nitropropionic acid (20 mg/kg i.p./day for 4 days) in striatal and cortical synaptosomes of Wistar rats was studied. The effects of 3-nitropropionic acid were evaluated as changes in the quantity of protein carbonyl groups, lipid peroxidation products, superoxide distumase activity and reduced succinate dehydrogenase activity. All changes were prevented by the pre-injection of nicotine (1.5 mg/kg i.p./day), beginning 4 days before and continuing for 4 days after the first injection of 3-nitropropionic acid. These findings indicate that: (i) 3-nitropropionic acid induces a state of oxidative stress in cortical and striatal synaptosomes and (ii) nicotine prevents oxidative stress induced by 3 nitropropinonic acid. In conclusion, the results show the ability of nicotine to modify neural response to 3-nitropropionic acid with the protective mechanism likely involving the antioxidative processes of nicotine. PMID- 15541419 TI - Increased release of immunoreactive dynorphin A1-17 from the spinal cord after intrathecal treatment with endomorphin-2 in anesthetized rats. AB - We previously demonstrated pretreatment with antiserum against dynorphin A1-17 attenuates endomorphin-2-induced analgesia and antianalgesia, suggesting that these endomorphin-2 effects are mediated by the release of dynorphin A1-17. Lumbar-cisternal spinal perfusion was used to measure the release of immunoreactive dynorphin A1-17 into spinal perfusates from urethane-anesthetized rats following endomorphin-2 or endomorphin-1 treatment within the perfusion solution. Treatment with endomorphin-2 (5-50 nmol) for 3 min caused a dose dependent increase of immunoreactive dynorphin A1-17 in spinal perfusates, with a maximal increase detected between 24 and 48 min after endomorphin-2 treatment, while levels returned to baseline within 60 min. Endomorphin-2-induced release of immunoreactive dynorphin A1-17 was attenuated by pretreatment with mu-opioid receptor antagonist naloxone or 3-methoxynaltrexone. Endomorphin-1 induced a slight increase in immunoreactive dynorphin1-17 as well, but only at the highest dose used (50 nmol). Our results suggest that endomorphin-2 stimulated a specific subtype of mu-opioid receptor to induce the release of immunoreactive dynorphin A1-17 in spinal cords of rats. PMID- 15541420 TI - Effects of chlorogenic acid and its metabolites on the sleep-wakefulness cycle in rats. AB - The effect of chlorogenic acid on the sleep-wakefulness cycle in rats was investigated in comparison with those of caffeic acid (the metabolite of chlorogenic acid) and dihydrocaffeic acid (the metabolite of caffeic acid). A significant prolongation of sleep latency was observed with chlorogenic acid and caffeic acid at a dose of 500 and 200 mg/kg, respectively. On the other hand, no remarkable effects were observed with dihydrocaffeic acid even at a dose of 500 mg/kg. Caffeine caused a significant increase in sleep latency and waking time and decrease in non-rapid eye movement sleep time at a dose of 10 mg/kg. In contrast, chlorogenic acid and its metabolites had no significant effects on each sleep state. From these results, it may be concluded that chrologenic acid caused a mild arousal effect compared with that of caffeine, and the effect of chlorogenic acid may have occurred through its metabolite caffeic acid. PMID- 15541421 TI - Effect of agmatine on the development of morphine dependence in rats: potential role of cAMP system. AB - Agmatine is an endogenous amine derived from arginine that potentiates morphine analgesia and blocks symptoms of naloxone-precipitated morphine withdrawal in rats. In this study, we sought to determine whether treatment with agmatine during the development of morphine dependence inhibits the withdrawal symptoms and that the effect is mediated by cAMP system. Exposure of rats to morphine for 7 days resulted in marked naloxone-induced withdrawal symptoms and agmatine treatment along with morphine significantly decreasing the withdrawal symptoms. The levels of cAMP were markedly increased in morphine-treated rat brain slices when incubated with naloxone and this increase was significantly reduced in rats treated with morphine and agmatine. The induction of tyrosine hydroxylase after morphine exposure was also reduced in locus coeruleus when agmatine was administered along with morphine. We conclude that agmatine reduces the development of dependence to morphine and that this effect is probably mediated by the inhibition of cAMP signaling pathway during chronic morphine exposure. PMID- 15541422 TI - The High-Ethanol Preferring rat as a model to study the shift between alcohol abuse and dependence. AB - The High-Ethanol Preferring line of rats (HEP), recently selected by R.D. Myers, is characterised by a high voluntary consumption of alcohol (3-4 g/kg/day for males and 6-8 g/kg/day for females, when a 10% ethanol solution is available as a choice vs. water) and a high sensitivity to taste reinforcement (saccharin, quinine). Our previous data obtained with HEP rats showed no evidence of development of dependence after long-term sustained alcohol intake. In this study, we subjected these rats to several long-term administration protocols suggested to favour the development of alcohol dependence, including multiple alcohol concentrations or sweetened alcohol solutions (ethanol 10% or 20%+saccharin), and deprivation periods. The results showed no increase in alcohol consumption, no shift of preference for alcohol solutions when offered as a free choice vs. a preferred saccharin solution, and a very limited alcohol deprivation effect when alcohol is made available after a period of deprivation, the three criteria used to demonstrate the development of dependence. Regardless of the method used, HEP rats failed to show dependence after long-term, heavy ethanol consumption. Resistance to ethanol dependence may in fact be genetically influenced and the HEP rat appears as a valuable model to search for factors involved in the transition from alcohol abuse to dependence. PMID- 15541423 TI - Involvement of dopamine receptors in the anti-immobility effects of dopamine re uptake inhibitors in the forced swimming test. AB - The effects of dopamine re-uptake inhibitors, bupropion and nomifensine on immobility in the forced swimming test were studied in mice. Bupropion and nomifensine reduced immobility time dose-dependently. Both drugs significantly displayed anti-immobility effects at doses without altering locomotor activity. Anti-immobility effects of bupropion and nomifensine were inhibited by the dopamine D1 receptor antagonist R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl 2,3,4,5-tetrahydro-1H-3-benzazepine-HCl (SCH 23390) and the dopamine D2 receptor antagonist sulpiride. These findings suggest that dopamine may be related to depression and dopamine D1 and dopamine D2 receptors play a role in the effects of dopamine re-uptake inhibitors. PMID- 15541424 TI - In vivo effectiveness of CGP7930, a positive allosteric modulator of the GABAB receptor. AB - The present study was aimed at assessing the in vivo effectiveness of the positive allosteric modulator of the gamma-aminobutyric acidB (GABAB) receptor, CGP7930 [2,6-di-tert-butyl-4-(3-hydroxy-2,2-dimethyl-propyl)-phenol]. The synergistic potentiation of GABAB receptor functioning, previously observed in different in vitro assays, has been confirmed in the present work, where pretreatment with CGP7930 (10-170 mg/kg, i.p.) resulted in a marked potentiation of the sedative/hypnotic effect of the GABAB receptor agonists, baclofen (40 mg/kg, i.p.) and gamma-hydroxybutyric acid (500 mg/kg, i.p.), in DBA mice. Pretreatment with the GABAB receptor antagonist, SCH 50911 [(S)-5,5-dimethyl-2 morpholine acetic acid; 100 mg/kg, i.p.], resulted in a complete blockade of the sedative/hypnotic effect of the combination of CGP7930 with either baclofen or gamma-hydroxybutyric acid. These results confirm that CGP7930 may constitute an interesting tool for pharmacological studies in the GABAB receptor field. PMID- 15541425 TI - Local transcutaneous electrical stimulation (TENS) effects in experimental inflammatory edema and pain. AB - Few studies in the literature associated transcutaneous electrical stimulation (TENS) use with an antiinflammatory activity. The purpose of this study was to investigate the effects of low (10 Hz)- and high (130 Hz)-frequency TENS on hyperalgesia and edema that occur after injection of carrageenan in rat paw. After induction of inflammation, either low- or high-frequency TENS was applied in the rat paw for 20 min, and the effect of TENS treatment on escape or paw withdrawal and edema was measured. Both low- and high-frequency TENS inhibited by 100% the hyperalgesia but not the edema response. However, low-frequency TENS presented longer lasting effect as compared with high-frequency TENS. Naltrexone treated animals showed a complete reversion of the analgesic effect induced by low- but not high-frequency TENS. Thus, our data demonstrated absence of an antiinflammatory effect associated to TENS use and confirmed the participation of endogenous opioids on low TENS-induced analgesia. PMID- 15541426 TI - Pharmacological profile of CR3465, a new leukotriene CysLT1 receptor antagonist with broad anti-inflammatory activity. AB - CR3465 (L-Tyrosine, N-[(2-quinolinyl)carbonyl]-O-(7-fluoro-2-quinolinylmethyl) sodium salt) is a potent antagonist of [3H]leukotriene D4 ([3H]LTD4) binding to guinea pig lung preparations, its Ki (4.7+/-0.7 nM) being comparable with that of montelukast (5.6+/-0.6 nM). In tracheal strips from standard or ovalbumin sensitized guinea pigs, CR3465 caused parallel rightward shifts in the concentration-response curves obtained with either LTD4 or antigen (pA(2), 8.74 and 8.15). Intravenous (i.v.) administration of the agent both antagonized (ED50, 9.9+/-1.9 microg/kg) and reverted LTD4 -induced bronchoconstriction of anesthetized guinea pigs. CR3465 reduced inflammatory infiltrates in the bronchoalveolar lavage fluid after antigen challenge of sensitized animals, and proved also active in inhibiting phosphodiesterase 3 (PDE3) and phosphodiesterase 4 (PDE4) activities exhibited by human platelets and neutrophils (IC50, 2.01+/ 0.07 and 4.7+/-0.5 microM). In line with properties shown by phosphodiesterase inhibitors, CR3465 reduced the contractile response of guinea pig airways to histamine and decreased N-formyl-Met-Leu-Phe (fMLP)-induced degranulation of human neutrophils (IC50, 13.8 microM). Oral administration (20 mg/kg) of the compound in rats produced a significant (37%) ex vivo inhibition of tumor necrosis factor-alpha (TNF-alpha) release from lipopolysaccharide-stimulated whole blood. Pharmacokinetic data in the rat demonstrated approximately 100% bioavailability of the agent. We conclude that CR3465 represents a potent leukotriene CysLT1 receptor antagonist with enhanced effects, being also useful for counteracting spasmogenic and inflammatory stimuli other than those elicited by cysteinyl-leukotrienes (Cys-LTs). PMID- 15541427 TI - Preimplantation genetic diagnosis--for or against humanity? PMID- 15541428 TI - How, where, and when should trial data be reported? PMID- 15541429 TI - Vaccination against human papillomaviruses shows great promise. PMID- 15541430 TI - Surgery for basal-cell carcinoma of the face. PMID- 15541431 TI - The metabolic vicious cycle in heart failure. PMID- 15541432 TI - Health research: what happened to the charcoal? PMID- 15541433 TI - Platelet transfusion and adverse outcomes. PMID- 15541434 TI - A novel drug target in Alzheimer's disease. PMID- 15541435 TI - Complex regional pain syndromes--how do we escape the diagnostic trap? PMID- 15541436 TI - Hearts and minds, from Darfur to locusts. PMID- 15541437 TI - A difficult ethics issue. PMID- 15541438 TI - A difficult ethics issue. PMID- 15541439 TI - Chlorproguanil-dapsone for malaria. PMID- 15541440 TI - Malaria initiative. PMID- 15541442 TI - Chlorproguanil-dapsone for malaria. PMID- 15541444 TI - Importance of protection of antimalarial combination therapies. PMID- 15541445 TI - Obstacles to health information for all. PMID- 15541446 TI - Obstacles to health information for all. PMID- 15541448 TI - Efficacy of a bivalent L1 virus-like particle vaccine in prevention of infection with human papillomavirus types 16 and 18 in young women: a randomised controlled trial. AB - BACKGROUND: Vaccination against the most common oncogenic human papillomavirus (HPV) types, HPV-16 and HPV-18, could prevent development of up to 70% of cervical cancers worldwide. We did a randomised, double-blind, controlled trial to assess the efficacy, safety, and immunogenicity of a bivalent HPV-16/18 L1 virus-like particle vaccine for the prevention of incident and persistent infection with these two virus types, associated cervical cytological abnormalities, and precancerous lesions. METHODS: We randomised 1113 women between 15-25 years of age to receive three doses of either the vaccine formulated with AS04 adjuvant or placebo on a 0 month, 1 month, and 6 month schedule in North America and Brazil. Women were assessed for HPV infection by cervical cytology and self-obtained cervicovaginal samples for up to 27 months, and for vaccine safety and immunogenicity. FINDINGS: In the according-to-protocol analyses, vaccine efficacy was 91.6% (95% CI 64.5-98.0) against incident infection and 100% against persistent infection (47.0-100) with HPV-16/18. In the intention-to-treat analyses, vaccine efficacy was 95.1% (63.5-99.3) against persistent cervical infection with HPV-16/18 and 92.9% (70.0-98.3) against cytological abnormalities associated with HPV-16/18 infection. The vaccine was generally safe, well tolerated, and highly immunogenic. INTERPRETATION: The bivalent HPV vaccine was efficacious in prevention of incident and persistent cervical infections with HPV-16 and HPV-18, and associated cytological abnormalities and lesions. Vaccination against such infections could substantially reduce incidence of cervical cancer. PMID- 15541449 TI - Surgical excision vs Mohs' micrographic surgery for basal-cell carcinoma of the face: randomised controlled trial. AB - BACKGROUND: Rates of recurrence of facial basal-cell carcinoma are consistently lower after Mohs' micrographic surgery (MMS) than after other treatments, such as surgical excision (SE). However, MMS is more time-consuming and therefore more expensive than SE. We investigated the types of facial basal-cell carcinomas in which MMS was more effective than SE. METHODS: 408 primary and 204 recurrent facial carcinomas (374 and 191 patients, respectively) were analysed separately in this prospective randomised study. Patients were assigned SE or MMS (each 204 primary, 102 recurrent), and received treatment at two hospitals in the Netherlands. The primary outcome was recurrence of carcinoma. Analysis was by intention to treat. FINDINGS: Of the basal-cell carcinomas included in the trial, 397 primary (198 MMS, 199 SE) and 201 recurrent (99, 102) tumours were actually treated. Of patients with primary carcinomas, 21 had both MMS and SE on different tumours. Nine with recurrent carcinomas had both treatments on different skin tumours. 66 primary and 13 recurrent carcinomas were lost to follow-up. Of the primary carcinomas, five (3%) recurred after SE compared with three (2%) after MMS during 30 months of follow-up. Of the recurrent carcinomas, three (3%) recurred after SE and none after MMS during 18 months of follow-up. Four recurrent carcinomas randomly assigned to the SE group were treated with MMS. Although both differences favoured MMS, they were not significant (primary, difference 1% [95% CI -2.5% to 3.7%], p=0.724; recurrent, 3.2% [-2.0% to 5.0%], p=0.119). Total operative costs of MMS were higher than those of SE (primary 405.79 Euros vs 216.86 Euros, recurrent 489.06 Euros vs 323.49 Euros; both p<0.001). INTERPRETATION: No definitive conclusion on recurrence rates of primary or recurrent basal-cell carcinomas is yet possible. Although recurrence rates were lower after MMS than after SE, the differences were not significant. PMID- 15541450 TI - The United Kingdom Infantile Spasms Study comparing vigabatrin with prednisolone or tetracosactide at 14 days: a multicentre, randomised controlled trial. AB - BACKGROUND: Infantile spasms, which comprise a severe infantile seizure disorder, have a high morbidity and are difficult to treat. Hormonal treatments (adrenocorticotropic hormone and prednisolone) have been the main therapy for decades, although little evidence supports their use. Vigabatrin has been recorded to have a beneficial effect in this disorder. We aimed to compare the effects of vigabatrin with those of prednisolone and tetracosactide in the treatment of infantile spasms. METHODS: The United Kingdom Infantile Spasms Study assessed these treatments in a multicentre, randomised controlled trial in 150 hospitals in the UK. The primary outcome was cessation of spasms on days 13 and 14. Minimum doses were vigabatrin 100 mg/kg per day, oral prednisolone 40 mg per day, or intramuscular tetracosactide depot 0.5 mg (40 IU) on alternate days. Analysis was by intention to treat. FINDINGS: Of 208 infants screened and assessed, 107 were randomly assigned to vigabatrin (n=52) or hormonal treatments (prednisolone n=30, tetracosactide n=25). None was lost to follow-up. Proportions with no spasms on days 13 and 14 were: 40 (73%) of 55 infants assigned hormonal treatments (prednisolone 21/30 [70%], tetracosactide 19/25 [76%]) and 28 (54%) of 52 infants assigned vigabatrin (difference 19%, 95% CI 1%-36%, p=0.043). Two infants allocated tetracosactide and one allocated vigabatrin received prednisolone. Adverse events were reported in 30 (55%) of 55 infants on hormonal treatments and 28 (54%) of 52 infants on vigabatrin. No deaths were recorded. INTERPRETATION: Cessation of spasms was more likely in infants given hormonal treatments than those given vigabatrin. Adverse events were common with both treatments. PMID- 15541451 TI - Prevention of cold-associated acute inflammation in familial cold autoinflammatory syndrome by interleukin-1 receptor antagonist. AB - BACKGROUND: Familial cold autoinflammatory syndrome (FCAS) is an autosomal dominant disorder characterised by recurrent episodes of rash, arthralgia, and fever after cold exposure. The genetic basis of this disease has been elucidated. Cryopyrin, the protein that is altered in FCAS, is one of the adaptor proteins that activate caspase 1, resulting in release of interleukin 1. METHODS: An experimental cold challenge protocol was developed to study the acute inflammatory mechanisms occurring after a general cold exposure in FCAS patients and to investigate the effects of pretreatment with an antagonist of interleukin 1 receptor (IL-1Ra). ELISA, real-time PCR, and immunohistochemistry were used to measure cytokine responses. FINDINGS: After cold challenge, untreated patients with FCAS developed rash, fever, and arthralgias within 1-4 h. Significant increases in serum concentrations of interleukin 6 and white-blood-cell counts were seen 4-8 h after cold challenge. Serum concentrations of interleukin 1 and cytokine mRNA in peripheral-blood leucocytes were not raised, but amounts of interleukin 1 protein and mRNA were high in affected skin. IL-1Ra administered before cold challenge blocked symptoms and increases in white-blood-cell counts and serum interleukin 6. INTERPRETATION: The ability of IL-1Ra to prevent the clinical features and haematological and biochemical changes in patients with FCAS indicates a central role for interleukin 1beta in this disorder. Involvement of cryopyrin in activation of caspase 1 and NF-kappaB signalling suggests that it might have a role in many chronic inflammatory diseases. RELEVANCE TO PRACTICE: These findings support a new therapy for a disorder with no previously known acceptable treatment. They also offer insights into the role of interleukin 1beta in more common inflammatory diseases. PMID- 15541452 TI - Uncoupling proteins in human heart. AB - Abnormal energetic activity in heart failure correlates inversely with plasma free-fatty-acid concentrations. However, the link between energetic and metabolic abnormalities is unknown. To investigate this association, we obtained blood samples from 39 patients undergoing coronary artery bypass graft surgery. Patients fasted overnight before samples were taken. When plasma free-fatty-acid concentrations were raised, cardiac mitochondrial uncoupling proteins (UCP) increased (isoform UCP2, p<0.0001; isoform UCP3, p=0.0036) and those of glucose transporter (GLUT4) protein decreased (cardiac, p=0.0001; skeletal muscle, p=0.0006). Consequently, energy deficiency in heart failure might result from increased mitochondrial UCPs (ie, less efficient ATP synthesis) and depleted GLUT4 (ie, reduced glucose uptake). New treatment to correct these energy defects would be to simultaneously lower plasma free fatty acids and provide an alternative energy source. PMID- 15541453 TI - Endometriosis. AB - Endometriosis is an oestrogen-dependent disorder that can result in substantial morbidity, including pelvic pain, multiple operations, and infertility. New findings on the genetics, the possible roles of the environment and the immune system, and intrinsic abnormalities in the endometrium of affected women and secreted products of endometriotic lesions have given insight into the pathogenesis of this disorder and serve as the background for new treatments for disease-associated pain and infertility. Affected women are at higher risk than the general female population of developing ovarian cancer, and they also may be at increased risk of breast and other cancers as well as autoimmune and atopic disorders. Clinicians should assess and follow up affected women for these and other associated disorders. There will probably be a new repertoire of approaches for treatment and perhaps cure of this enigmatic disorder in the near future. PMID- 15541454 TI - Coping with endometriosis. PMID- 15541455 TI - Lessons learned from complex emergencies over past decade. AB - Major advances have been made during the past decade in the way the international community responds to the health and nutrition consequences of complex emergencies. The public health and clinical response to diseases of acute epidemic potential has improved, especially in camps. Case-fatality rates for severely malnourished children have plummeted because of better protocols and products. Renewed focus is required on the major causes of death in conflict affected societies--particularly acute respiratory infections, diarrhoea, malaria, measles, neonatal causes, and malnutrition--outside camps and often across regions and even political boundaries. In emergencies in sub-Saharan Africa, particularly southern Africa, HIV/AIDS is also an important cause of morbidity and mortality. Stronger coordination, increased accountability, and a more strategic positioning of non-governmental organisations and UN agencies are crucial to achieving lower maternal and child morbidity and mortality rates in complex emergencies and therefore for reaching the UN's Millennium Development Goals. PMID- 15541456 TI - Delayed graft function in kidney transplantation. AB - Delayed graft function is a form of acute renal failure resulting in post transplantation oliguria, increased allograft immunogenicity and risk of acute rejection episodes, and decreased long-term survival. Factors related to the donor and prerenal, renal, or postrenal transplant factors related to the recipient can contribute to this condition. From experimental studies, we have learnt that both ischaemia and reinstitution of blood flow in ischaemically damaged kidneys after hypothermic preservation activate a complex sequence of events that sustain renal injury and play a pivotal part in the development of delayed graft function. Elucidation of the pathophysiology of renal ischaemia and reperfusion injury has contributed to the development of strategies to decrease the rate of delayed graft function, focusing on donor management, organ procurement and preservation techniques, recipient fluid management, and pharmacological agents (vasodilators, antioxidants, anti-inflammatory agents). Several new drugs show promise in animal studies in preventing or ameliorating ischaemia-reperfusion injury and possibly delayed graft function, but definitive clinical trials are lacking. The goal of monotherapy for the prevention or treatment of is perhaps unattainable, and multidrug approaches or single drug targeting multiple signals will be the next step to reduce post-transplantation injury and delayed graft function. PMID- 15541457 TI - When head rules heart. PMID- 15541458 TI - Calcium channels, transporters and exchangers in placenta: a review. AB - Calcium (Ca2+) entry in cells is crucial for development and physiology of virtually all cell types. It acts as an intracellular (second) messenger to regulate a diverse array of cellular functions, from cell division and differentiation to cell death. Among candidates for Ca2+ entry in cells are voltage-dependant Ca2+ channels (VDCCs), transient receptor potential (TRP) related Ca2+ channels and store-operated Ca2+ (SOC) channels. Plasma membrane Ca2+-ATPases (PMCA) and Na+/Ca2+ exchanger (NCX) are mainly responsible for Ca2+ extrusion. These different Ca2+channels/transporters and exchangers exhibit specific distribution and physiological properties. During pregnancy, the syncytiotrophoblast layer of the human placenta transfers as much as 30 g of Ca2+ from the mother to the fetus, especially in late gestation where Ca2+ transport through different channels must increase in response to the demands of accelerating bone mineralization of the fetus. The identification and characterization of the different Ca2+ channels/transporters and exchangers on the brush-border membrane (BBM) facing the maternal circulation, and the basal plasma membrane (BPM) facing the fetal circulation; placental membrane of the syncytiotrophoblasts have been the focus of numerous studies. This review discusses current views in this field regarding localization and functions during transcellular Ca2+ entry and extrusion from cells particularly in the placenta. PMID- 15541459 TI - Calyculin-A disrupts subplasmalemmal junction and recurring Ca2+ waves in vascular smooth muscle. AB - Excitation-contraction coupling (E-C coupling) in phenylephrine(PE)-stimulated rabbit inferior vena cava (IVC) depends on the generation of asynchronous recurring Ca2+ waves in the in situ vascular smooth muscle cells (VSMC). Previous studies by our group have implicated a putative non-selective cationic store operated channel and the reverse-mode Na+-Ca2+ exchange in refilling of the intracellular Ca2+ store via the sarco/endoplasmic reticulum ATPase (SERCA) and the maintenance of the recurring Ca2+ waves. We hypothesize that for the proper functioning of these three Ca2+ translocators in the process of SR refilling, the plasma membrane (PM) and the underlying superficial sarcoplasmic reticulum (SR) form specialized PM-SR junctions, which are essential for the maintenance of the recurring Ca2+ waves. In order to test this hypothesis, calyculin-A, a serine/threonine phosphatase inhibitor that has been demonstrated to result in the disruption of the PM-SR junctions was used. In the control rabbit IVC, electron microscopy of the in situ VSMC indicates that 14.2+/-0.7% of the PM is closely apposed by the prominent superficial SR network, forming numerous flattened PM-SR junctional cytoplasmic spaces. In the control IVC stimulation with 5 microM PE resulted in sustained recurring Ca2+ waves with a frequency of 0.42+/-0.02 Hz. In calyculin-A treated rabbit IVC, a concentration-dependent dissociation of the superficial SR and loss of PM-SR junctions was observed. This progressive loss of the PM-SR junctions occurs over the same concentration range as the inhibition of PE-induced recurring Ca2+ waves. These findings offer support for the hypothesis that the presence of the PM-SR junctions is required for the generation of asynchronous recurring Ca2+ waves, which underlie excitation-contraction coupling in the VSMC of the rabbit IVC. PMID- 15541460 TI - Control of SERCA2a Ca2+ pump mRNA stability by nuclear proteins: role of domains in the 3'-untranslated region. AB - Alternative splicing of the sarco/endoplasmic reticulum (SERCA2) Ca2+ pump transcript generates the two isoforms: SERCA2a in left ventricular myocytes (LVM) and SERCA2b in most tissues. Nuclear protein extracts from left ventricular myocytes can cause a decay of the 3'-region of the SERCA2a. To determine if all the domains in the 800 b SERCA2a 3'-end region (3344-4243) are equally stable, we examined in vitro decay of synthetically capped, polyadenylated overlapping RNA fragments 2A1-2A6 from the 3'-end region of SERCA2a. Whereas 2A1-2A5 RNAs were stable, the distal fragment 2A6 (4135-4243 b) decayed rapidly. Deleting the 2A6 sequence from the 800-b 3'-end region increased its stability. In mobility shift assays, 2A6 bound to protein(s) in the LVM nuclear extracts in a specific manner: unlabelled 2A6 or the 800 b 3'-region RNA competed for binding but poly A, poly U, and poly C RNA did not. Secondary structure analysis revealed three hairpin loops in 2A6. Experiments using small synthetic RNA fragments for competition with 2A6 binding to nuclear proteins were consistent with a model involving the three hairpin loops. Thus, the secondary structure of the distal domain of SERCA2a RNA may be important in regulating its stability. PMID- 15541462 TI - Prostaglandin F2alpha potentiates the calcium dependent activation of mitochondrial metabolism in luteal cells. AB - Cytoplasmic Ca2+ signals are transferred to the mitochondria and activate the Krebs cycle. We have compared the efficiency of this process for two Ca2+ mobilising agonists, PGF2alpha and ATP (acting at metabotropic P2 receptors) in rat luteal cells. [Ca2+]c, [Ca2+]m and mitochondrial NAD(P)H were monitored by means of microspectrofluorimetry and confocal microscopy. While both agonists caused similar elevations of [Ca2+]c, changes in NAD(P)H were larger in response to PGF2alpha than to ATP. PGF2alpha more effectively increased NAD(P)H level also in mouse luteal cells. PGF2alpha caused a faster rate of rise of NAD(P)H fluorescence than ATP when reoxidation was prevented with rotenone, suggesting a faster rate of NAD(P)+ reduction. The NAD(P)H response to both agonists was dependent on the mobilisation of stored Ca2+. We found no difference in the efficacy of transmission of the [Ca2+]c signal to mitochondria in response to PGF2alpha and ATP. Raising [Ca2+]c with ionomycin increased the NAD(P)H signal, which was further raised by PGF2alpha but not by ATP. These data suggest that PGF2alpha potentiates the Ca2+-induced stimulation of mitochondrial metabolism by a Ca2+-independent mechanism and shows that agonists may modulate mitochondrial function differentially through a novel process beyond the simple transfer of Ca2+ from ER to mitochondria. PMID- 15541461 TI - Intercellular communication: role of gap junctions in establishing the pattern of ATP-elicited Ca2+ oscillations and Ca2+-dependent currents in freshly isolated aortic smooth muscle cells. AB - Cytosolic-free [Ca2+] was evaluated in freshly dissociated smooth muscle cells from mouse thoracic aorta by the ratio of Fura Red and Fluo 4 emitted fluorescence using confocal microscopy. The role of intercellular communication in forming and shaping ATP-elicited responses was demonstrated. Extracellular ATP (250 microM) elicited [Ca2+]i transient responses, sustained [Ca2+]i rise, periodic [Ca2+]i oscillations and aperiodic repetitive [Ca2+]i transients. Quantity of smooth muscle cells in the preparation responding to ATP with periodical [Ca2+]i oscillations depended on the density of isolated cells on the cover slip. ATP-elicited bursts of [Ca2+]i spikes in 66+/-7% of cells in dense and in 33+/-8.5% of cells in non-dense preparations. The number of cells responding to ATP with bursts of [Ca2+]i spikes decreased from 55+/-5% (n=84) to 14+/-3% (n=141) in dense preparations pretreated with carbenoxolone. Simultaneous measurement of [Ca2+]i and ion currents revealed a correlation between [Ca2+]i and current oscillations. ATP-elicited bursts of current spikes in 76% of cells regrouped in small clusters and in 9% of isolated cells. Clustered cells responding to ATP with current oscillations had higher membrane capacity than clustered cells with transient and sustained ATP-elicited responses. Lucifer Yellow (1% in 130 mM KCl) injected into one of clustered cells was transferred to the neighboring cell only when ATP-elicited oscillations. Fast application of carbenoxolone (100 microM) inhibited ATP (250 microM) elicited Ca2+-dependent current oscillations. Taken together these results suggest that the probability of ATP (250 microM) triggered cytosolic [Ca2+]i oscillations accompanied with K+ and Cl- current oscillations increased with the coupling of smooth muscle cells. PMID- 15541463 TI - Ca2+ clearance mechanisms in neurohypophysial terminals of the rat. AB - The importance of intracellular calcium ([Ca2+]i) in the release of vasopressin (AVP) and oxytocin from the central nervous system neurohypopyhysial nerve terminals has been well-documented. To date, there is no clear understanding of Ca2+ clearance mechanisms and their interplay with transmembrane Ca2+ entry, intracellular [Ca2+]i transients, cytoplasmic Ca2+ stores and hence the release of AVP at the level of a single nerve terminal. Here, we studied the mechanism of Ca2+ clearance in freshly isolated nerve terminals of the rat neurohypophysis using Fura-2 Ca2+ imaging and measured the release of AVP by radioimmuno assay. An increase in the K+ concentration in the perfusion solution from 5 to 50 mM caused a rapid increase in [Ca2+]i and AVP release. Returning K+ concentration to 5 mM led to rapid restoration of both responses to basal level. The K+-evoked [Ca2+]i and AVP increase was concentration-dependent, reliable, and remained of constant amplitude and time course upon successive applications. Extracellular Ca2+ removal completely abolished the K+-evoked responses. The recovery phase was not affected upon replacement of NaCl with sucrose or drugs known to act on intracellular Ca2+ stores such as thapsigargin, cyclopiazonic acid, caffeine or a combination of caffeine and ryanodine did not affect either resting or K+-evoked [Ca2+]i or AVP release. By contrast, the plasma membrane Ca2+ pump inhibitor, La3+, markedly slowed down the recovery phase. The mitochondrial respiration uncoupler, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), slightly but significantly increased the basal [Ca2+]i, and also slowed down the recovery phase of both [Ca2+]i and release responses. In conclusion, we show in nerve terminals that (i) Ca2+ extrusion through the Ca2+ pump in the plasma membrane plays a major role in the Ca2+ clearance mechanisms of (ii) Ca2+ uptake by mitochondria also contributes to the Ca2+ clearance and (iii) neither Na+/Ca2+ exchangers nor Ca2+ stores are involved in the Ca2+ clearance or in the maintenance of basal [Ca2+]i or release of AVP. PMID- 15541464 TI - Developmental control of cAMP-induced Ca2+-influx by cGMP: influx is delayed and reduced in a cGMP-phosphodiesterase D deficient mutant of Dictyostelium discoideum. AB - It was previously shown that cGMP enhances cAMP-induced Ca2+-influx in Dictyostelium discoideum. This finding is based on experiments done with strains defective in cGMP-hydrolysis, the streamer F cells. In this work, we show that these chemically mutagenized cells display different properties in their cAMP induced light-scattering response and cAMP-induced Ca2+-influx compared with a cGMP-phosphodiesterase knock-out strain, pdeD KO, generated by homologous recombination. PdeD KO cells possess a reduced Ca2+-influx that is developmentally regulated. This finding contradicts the result of streamer F cells, where cAMP-induced Ca2+-influx is prolonged and elevated. Both mutants, however, showed a three to four-fold delayed response to cAMP at 3-4h of starvation. Thus, the consequence of an elevated cGMP concentration is a delay and an inhibition of Ca2+-influx and not an enhancement. Results obtained with streamer F cells should therefore be interpreted with caution because the mutation(s) responsible for the divergent phenotype to pdeD KO cells has not been identified. We show by the use of membrane-permeant cGMP-analogues in wild type (wt) cells, permeabilized cells and measurements on isolated vesicles that the cause for the reduced Ca2+-influx seems to be due to developmentally regulated Ca2+-channel inhibition by cGMP. PMID- 15541465 TI - Inhibition of the Na+-H+ exchanger with cariporide abolishes stretch-induced calcium but not sodium accumulation in mouse ventricular myocytes. AB - We address the question whether activation of the sodium-proton exchanger (NHE) does contribute to the stretch-induced accumulation of intracellular sodium and calcium in mouse ventricular myocytes. NHE-blocker cariporide (10 microM) were applied to the bath for 10 min. Axial stretch was applied for 2 min by increasing the distance between an adherent glass stylus and the patch pipette by 20%. Myocytes (stimulated at 3 Hz) were shock-frozen in diastole and the membrane currents monitored till cryofixation. Controls were treated identically, but not stretched. Total sodium and calcium concentrations ([Na], [Ca]=sum of free and bound Na and Ca) were measured by electron probe microanalysis (EPMA) in peripheral and central cytosol, mitochondria, nucleus and nuclear envelope. Cariporide did not reduce the stretch-activated negative current. The stretch induced rise in [Na] was not different in the presence and in the absence of cariporide. Cariporide significantly reduced diastolic [Ca] in the cytosol of stretched myocytes. Since cariporide does not prevent the stretch-induced [Na] accumulation, we suggest that not NHE but the stretch-activated streptomycin sensitive current I(SAC) causes the well documented stretch-induced [Na] accumulation. The discovery that cariporide prevents the stretch-induced rise in cytosolic [Ca] demonstrates an important additional effect of the drug on calcium handling. PMID- 15541466 TI - Role of the nicotinic acid group in NAADP receptor selectivity. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) has been shown to be an intracellular Ca2+-releasing messenger in a wide variety of systems to date. Its actions are both potent and highly specific despite differing structurally from the endogenous cellular co-factor and its precursor, NADP, only in the substitution of a hydroxyl for the amine group at the 3' position of the pyridine ring. This substitution allows NAADP to bind to a membrane-localized binding site in sea urchin egg homogenates with an IC50 at least 1000-fold greater than that of NADP as measured by competition radioligand binding assays. This suggests that the NAADP receptor protein must include certain features in the NAADP binding site that regulate this specificity. In order to investigate this interaction, we synthesised a series of NAADP analogues differing from NAADP at the 3' position of the pyridine ring that included both simple carboxylic acid analogues as well as a series of chemical isosters. We then investigated both their affinity for the NAADP binding site in sea urchin egg homogenates and their ability to activate the NAADP sensitive Ca2+ channel. We hereby show that a negative charge at the 3' position is an important determinant of affinity but the protein displays a large tolerance for the size of the group. Furthermore, the protein does not easily accommodate multiple charged groups or large uncharged groups. PMID- 15541467 TI - Nitroxyl triggers Ca2+ release from skeletal and cardiac sarcoplasmic reticulum by oxidizing ryanodine receptors. AB - The biological activity of nitric oxide (NO) and NO-donors has been extensively investigated yet few studies have examined those of nitroxyl (HNO) species even though both exist in chemical equilibrium but oxidize thiols by different reaction mechanisms: S-nitrosation versus disulfide bond formation. Here, sodium trioxodinitrate (Na2N2O3; Angeli's salt; ANGS) was used as an HNO donor to investigate its effects on skeletal (RyR1) and cardiac (RyR2) ryanodine receptors. At steady-state concentrations of nanomoles/L, HNO induced a rapid Ca2+ release from sarcoplasmic reticulum (SR) vesicles then the reducing agent dithiothreitol (DTT) reversed the oxidation by HNO resulting in Ca2+ re-uptake by SR vesicles. With RyR1 channel proteins reconstituted in planar bilayers, HNO added to the cis-side increased the open probability (Po) from 0.056+/-0.026 to 0.270+/-0.102 (P<0.005, n=4) then DTT (3 mM) reduced Po to 0.096+/-0.040 (P<0.01, n=4). In parallel experiments, the time course of HNO production from ANGS was monitored by EPR and UV spectroscopy and compared with the rate of SR Ca2+ release indicating that picomolar concentrations of HNO triggered SR Ca2+ release. Controls showed that the hydroxyl radical scavenger, phenol did not alter ANGS-induced SR Ca2+ release, indicating that hydroxyl radical production from ANGS did not account for Ca2+ release from the SR. The findings indicate that HNO is a more potent activator of RyR1 than NO and that HNO activation of RyRs may contribute to NO's activation of RyRs and to the therapeutic effects of HNO-releasing prodrugs in heart failure. PMID- 15541468 TI - Gemtuzumab ozogamicin: a component of induction therapy in AML? PMID- 15541470 TI - Cancer chemotherapy reduces plasma total antioxidant capacity in children with malignancies. AB - The sum of endogenous and food-derived antioxidants provides an estimate of the total antioxidant capacity (TAC) of the extracellular fluids, while corrected TAC (cTAC) is an estimation of the exogenously provided antioxidants. Similar values for TAC and cTAC were observed between cancer free children and children with malignancy at diagnosis. Antineoplastic treatment induced a significant decrease of TAC and cTAC during chemotherapy. Additionally to the dietary factors, this might be attributed to the antineoplastic drugs as shown by the significant increase of ROS after administration of chemotherapeutic agents both in vitro and in vivo. According to our preliminary results TAC and cTAC returned to normal after the end of therapy. PMID- 15541469 TI - A phase-I clinical trial of active immunotherapy for acute leukemia using inactivated autologous leukemia cells mixed with IL-2, GM-CSF, and IL-6. AB - We evaluated the efficacy and toxicity of vaccination in 29 patients with relapsed or refractory acute leukemia using inactivated autologous leukemia cells combined with interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-6. MHC-I, MHC-II, and B7-1 expression status on the surface of leukemia cells and the cytokine profile of IFN-gamma and IL-10 in serum before and after vaccination was detected. RESULTS: Five achieved a complete remission (CR) and six a partial remission (PR) in this vaccination procedure. Adverse effects were erythema, swelling erosion, and even ulcers at vaccination sites and low grade fever during the first three days of vaccination. No other significant side effects were observed. The expression of MHC-I and MHC II on leukemia cells was 100% and 90% positive, respectively. B7-1 was exclusively expressed on some cases of M4 and M5. The efficacy of the vaccine was statistically associated with the expression status of B7-1 on leukemia cells (P < 0.01). The serum level of IL-10 reduced significantly in the five patients who achieved complete remission (CR) after vaccination as compared with when they were originally diagnosed (P < 0.01). CONCLUSION: We presented here a promising immunotherapy in the treatment of acute leukemia, especially for F.A.B. M5. PMID- 15541471 TI - Structural analysis and expression profile of a novel gene on chromosome 5q23 encoding a Golgi-associated protein with six splice variants, and involved within the 5q deletion of a Ph(-) CML patient. AB - We have identified a novel gene, upstream of the cytokine gene cluster region in 5q23-31, residing within one of the most common deleted segments associated with MDS. The novel gene exhibits significant alternative splicing generating at least six splice variants encoding four putative proline-rich protein isoforms, one of which is Golgi-associated. The gene is ubiquitously expressed and conserved among species with the C. elegans homologue being the most interesting, since it resides within an operon with two other genes, phospholipase D and dishevelled, a member of the Wnt pathway, suggesting a functional association. In addition, the novel gene and other key regulatory genes of the region, such IL3, Ril, AF5q31 and TCF-1, were found to be deleted in an atypical CML case, thus underscoring the significance of this subregion in the leukemogenesis process. PMID- 15541472 TI - Clinical features of prognostic significance in myelodysplastic patients with normal karyotype at high risk of transformation. AB - The International Prognostic Scoring System (IPSS) for myelodysplastic syndromes (MDS) has defined patients with a normal karyotype as a good risk cytogenetic subgroup, but nevertheless a fraction of these patients has a poor outcome similar to that of high risk patients. We retrospectively analysed our series of myelodysplastic patients with normal karyotype observed in a period of 11 years, with the aim of identifying clinical features of possible prognostic significance within this subgroup of patients. Multivariate analysis showed that among clinical scoring systems, the Bournemouth score appears the best prognostic indicator for risk of leukemic transformation, and platelet count <100 x 10(9)/l( 1), presence of haemorrhagic symptoms at time of diagnosis and morphologic FAB classification are the main prognostic factors for prediction of survival. In the absence of genetic abnormalities as detected by conventional cytogenetics or even the more sensitive molecular techniques in MDS, clinical variables could be of help in identifying patients with different prognosis, suitable for risk adapted therapeutic strategies. PMID- 15541473 TI - Relation between bone marrow angiogenesis and serum levels of angiogenin in patients with myelodysplastic syndromes. AB - Angiogenesis is implicated in the progression of myelodysplastic syndromes (MDS). Bone marrow microvascular density (MVD), serum angiogenin (ANG) and interleukin 6 (IL-6) were measured in 67 patients with untreated MDS. MVD, ANG and IL-6 were significantly higher in the patient group as a whole when compared to controls (P < 0.01). MVD and ANG were significantly higher in subtypes with a high-risk for leukemic transformation (RAEB, RAEB-t and CMML) than in low-risk subtypes (RA and RARS) (P < 0.01). In the MDS group, a positive correlation was found between ANG and IL-6 (P < 0.001) and also between MVD and IL-6 (P < 0.05). Using multivariate analysis, only IL-6 displayed independent prognostic value and was inversely related to MDS survival. PMID- 15541474 TI - Methylation analysis of the adenomatous polyposis coli (APC) gene in adult T-cell leukemia/lymphoma. AB - We investigated methylation status of the adenomatous polyposis coli (APC) gene in adult T-cell leukemia/lymphoma (ATL). APC methylation was found in 15 of 31 (48%) primary samples, and 2 of 4 (50%) ATL cell lines. Methylation of the APC gene occurred more frequently in acute ATL (12/21) (57%) than chronic ATL (1/8) (13%) (P = 0.03). APC was not expressed in the APC-methylated ATL cell line ST1. Demethylation with 5-azacytidine treatment restored APC expression in the ST1 cell line. Our data show that hypermethylation of the APC gene is involved in the pathogenesis of ATL. PMID- 15541475 TI - Phase II pilot trial of gemtuzumab ozogamicin (GO) as first line therapy in acute myeloid leukemia patients age 65 or older. AB - The prognosis of acute myeloid leukemia (AML) in older adults is generally poor. Standard cytotoxic chemotherapy is usually poorly tolerated and provides dismal results in this patient population. Gemtuzumab ozogamicin (GO) is an immunoconjugate that has activity in the relapsed and refractory setting. We hypothesized that administering this agent, as an initial treatment in AML patients over the age of 65 would have, at least, similar efficacy to standard therapy, but at much less toxicity. We report on the first 12 patients that were treated solely with GO as induction, consolidation, and maintenance therapy as part of a single institution clinical trial. The treatment was well tolerated, with a response rate of 27%. Toxicities were acceptable, although five patients (41%) developed cardiac toxicity three of which had grade 3 and/or 4. Responses were present regardless of the karyotype, and lasted for a median of 7.6 months. These early results suggest that the option of single agent monoclonal antibody therapy is viable in this patient population and that additional studies are warranted with this agent either alone or in combination as initial therapy. PMID- 15541476 TI - Germline mutations in Dok1 do not predispose to chronic lymphocytic leukemia. AB - The genetic basis of familial CLL is poorly understood and to date no gene which when mutated in the germline has been unambiguously shown to confer susceptibility to the disease. Dok1 maps to chromosome 2p13, a region commonly rearranged in CLL. Dok1 inhibits MAP kinase activity, down-regulates cell proliferation and has a suppressive effect on cellular transformation and B-cell signalling pathways. A recent report has implicated mutation of Dok1 in the aetiology of CLL. To examine the proposition that germline mutations in Dok1 act as high penetrance susceptibility alleles for CLL we screened 140 familial cases for functional sequence variants. No pathogenic mutations were detected. This result indicates that germline mutations in Dok1 are unlikely to cause an inherited predisposition to CLL. PMID- 15541477 TI - Interaction of bone marrow stromal cells with lymphoblasts and effects of predinsolone on cytokine expression. AB - Cytokines play a key role in the differentiation, growth and survival of hematopoietic cells in the bone marrow (BM) stroma microenvironment. The mechanisms by which stromal derangements may contribute to the evolution of hematopoietic neoplasias are largely unknown. Here, we characterized BM stromal cells isolated from children with acute lymphoblastic leukemia and determined the effect of the interaction between stromal cells and lymphoblasts on cytokine expression as well as the effect of prednisolone using mono- and co-culture models. The analyses demonstrate that (1) stromal cells and lymphoblasts display different patterns of cytokine gene expression individually. (2) Stromal cells influence gene expression of cytokines in lymphoblasts and vice versa. (3) Glucocorticoid substitution inhibit cytokine gene expression in stromal cells. These findings indicate that stromal cells are important components involved in malignant hematopoiesis and also in response to therapy. PMID- 15541478 TI - Normal neutrophil maturation is associated with selective loss of MAP kinase activation by G-CSF. AB - Although both GM-CSF and G-CSF activate p42/44 MAPK in neutrophil progenitors, the ability of G-CSF to cause MAPK activation is lost in mature neutrophils, while GM-CSF exposure still causes activation. The mechanism of this differential effect related to maturation status has not been explored. We verified that G-CSF and GM-CSF receptors remain functional on purified mature neutrophils by demonstrating that both cytokines caused phosphorylation of STAT3. However, only GM-CSF was capable of activating MAPK as assessed by gel shift and in vitro kinase assay. Both G-CSF and GM-CSF caused activation of p21 ras in neutrophils, demonstrating that early events in the ras-MAPK pathway remain functional after stimulation by either cytokine. Inhibition of tyrosine phosphatase activity by pervanadate restored the ability of G-CSF to activate MAPK in mature neutrophils. Specific inhibition of the SHP-1 phosphatase, known to be activated by G-CSF but not GM-CSF also restored the ability of G-CSF to activate MAPK in neutrophils. These studies suggest that G-CSF activation of SHP-1 may be an important regulatory step for permitting optimal terminal differentiation during neutrophil production and add to our knowledge of the instructional role of G-CSF and GM-CSF for balancing proliferation and differentiation of neutrophil progenitor cells. This information may prove useful for the understanding of conditions in which neutrophil proliferative/differentiative balancing is dysregulated, such as myeloid leukemia and myelodysplastic disorders. PMID- 15541479 TI - 17beta-Estradiol transcriptionally represses human insulin receptor gene expression causing cellular insulin resistance. AB - In this study, we demonstrate that 17beta-estradiol (E(2)) inhibits human insulin receptor (IR) gene expression in a dose- and time-dependent manner in U-937 human promonocytic cells. Using cells transfected with the -1819 to -271 bp fragment of the human IR promoter (wild type promoter) and treated with E(2), we show that this repression is regulated at the transcriptional level. The steroid was also found to diminish the insulin responsiveness of the cells in terms of cell survival, DNA synthesis, glucose transport, and glucose oxidation, this last effect possibly involving reduced phosphatidylinositol 3-kinase (PI3-kinase) activity. These data provide new information on the molecular mechanisms of estrogen-inducing insulin resistance in human cells. PMID- 15541480 TI - Negative regulatory elements are present in the human LMO2 oncogene and may contribute to its expression in leukemia. AB - Ectopic expression of LMO2 occurs in approximately 45% of T-lineage acute lymphoblastic leukemias (T-ALL), sometimes in association with chromosomal translocations. Recently, a lymphoproliferative disorder developed in two participants in a gene therapy trial due to LMO2 activation via integration of the retroviral vector. To investigate these regulatory disruptions, we analyzed the promoter region and identified a tissue-specific repressor. The fragment containing this element could also produce tissue-specific suppression of transcription from the SV40 promoter. This suppression involves histone acetylation which can be relieved with Trichostatin A (TSA). The negative element is in a region consistently removed from LMO2 in the known chromosomal translocations. PMID- 15541481 TI - Triptolide inhibits transcription factor NF-kappaB and induces apoptosis of multiple myeloma cells. AB - Triptolide has been reported to be effective in the treatment of auto-immune diseases. This study investigates the cytotoxic function of triptolide on multiple myeloma (MM) cells. We found that triptolide inhibited the proliferation of both RPMI8226 and U266 cells in a dose-dependent manner (10-80 ng/mL). Triptolide induced apoptosis in MM cells through activation of the cystein protease caspase 8, 9 and 3, and subsequent cleavage of the DNA repair enzyme poly (ADP-ribose) polymerase. Apoptosis was confirmed with cell-cycle analysis and annexin V staining. Moreover, triptolide down-regulated nuclear factor (NF) kappaB activity in MM cell lines. In addition, triptolide also induced chemosensitivity to doxorubicin and suppressed cell proliferation of fresh MM cells. Therefore, triptolide appears to be a potent inducer of apoptosis in myeloma cells, and might have some benefit in the treatment of myeloma patients. PMID- 15541482 TI - Rituximab in primary conjunctiva lymphoma. AB - Our experience with a patient with a primary conjunctiva lymphoma who was treated with conventional dose of rituximab obtaining a complete response without acute and late local toxicity for the eye. PMID- 15541483 TI - PCR for monoclonal gene rearrangements can differentiate infantile acute lymphoblastic leukemia from cytomegalovirus infection. PMID- 15541484 TI - Role of all-trans-retinoic acid (ATRA) in the consolidation therapy of acute promyelocytic leukaemia (APL). AB - The role of all-trans-retinoic acid (ATRA) in the consolidation therapy of acute promyelocytic leukaemia (APL) is undefined at present. Between 1994 and 1999, we treated 38 newly diagnosed APL patients with ATRA and chemotherapy during the induction and consolidation. ATRA was given for 28 days each during the induction and 2 cycles of consolidation therapy. Chemotherapy consisted of daunorubicin and cytarabine. No maintenance therapy was given. ATRA was well-tolerated during consolidation therapy with no cases of retinoic acid syndrome. The 5-year overall and leukaemia-free survival of study patients was 82% (95% C.I. 70-95%) and 78% (95% C.I. 65-93%), respectively. These data are comparable to the studies that used prolonged maintenance with ATRA +/- chemotherapy. The role of ATRA during consolidation therapy of APL merits further investigation as this may allow shortening the overall duration of APL treatment. PMID- 15541485 TI - Role of BCRP and its expression with MDR1 in adult AML. PMID- 15541486 TI - Methoxy-phenyl substituted ansa-titanocenes as potential anti-cancer drugs derived from fulvenes and titanium dichloride. AB - Starting from 6-(4'-methoxyphenyl)fulvene (1a), 6-(2',4',6' trimethoxyphenyl)fulvene (1b), or 6-(3',5'-dimethoxyphenyl)fulvene (1c), [1,2 di(cyclopentadienyl)-1,2-di(4'-methoxyphenyl)-ethanediyl] titanium dichloride (2a), [1,2-di(cyclopentadienyl)-1,2-bis(2',4',6'-trimethoxyphenyl)-ethanediyl] titanium dichloride (2b), and [1,2-di(cyclopentadienyl)-1,2-bis(3',5' dimethoxyphenyl)-ethanediyl] titanium dichloride (2c) were synthesised. When titanocenes 2a-c were tested against pig kidney carcinoma cells (LLC-PK) inhibitory concentrations (IC50) of 2.8 x 10(-4), 3.6 x 10(-4) and 2.1 x 10(-4) M, respectively, were observed. PMID- 15541487 TI - Reinvestigation of the copper(II)-carcinine equilibrium system: "two-dimensional" EPR simulation and NMR relaxation studies for determining the formation constants and coordination modes. AB - The equilibria and solution structure of complexes formed between copper(II) and carcinine (beta-alanyl-histamine) at 2< or = pH< or =11.2 have been studied by EPR and NMR relaxation methods. Beside the species that have already been described in the literature from pH-potentiometric measurements, several new complexes have been identified and/or structurally characterized. The singlet on the EPR spectrum detected in equimolar solutions at pH 7, indicates the formation of an oligomerized (CuL)n(2n+) complex, with [NH2,Nim] coordination. The oligomerization is probably associated with the low stability of the ten-membered macrochelate ring, which would form in the mononuclear complex CuL2+. In presence of moderate excess of ligand the formation of four new bis-complexes (CuL2Hn(2+n), n=2,1 and 0/-1) was detected with [Nim][Nim], [NH2,Nim][Nim] and [NH2,N-,Nim][Nim] type co-ordination modes, respectively. At higher excess of ligand ([L]/[Cu2+]>10) and at pH approximately 7, the predominant species is CuL4H2(4+). The 1H and 13C relaxation measurements of carcinine solutions (0.6 M) in presence of 0 mM< or = [Cu2+](tot)< or = 5 mM at pH=6.8, allowed us to extract the carbon-to-metal distances, the electronic relaxation and tumbling correlation times, as well as the ligand exchange rate for the species CuL4H2(4+). According to these results, the metal ion is [4Nim] co-ordinated in the equatorial plane, while the neutral amino groups are unbounded. Since naturally occurring carcinine shows in vivo antioxidant property, the SOD-like activity of the copper(II) carcinine system has also been investigated and the complex CuLH(-1) was found to be highly active. PMID- 15541488 TI - Metal-mediated formation of fibrillar ABri amyloid. AB - British amyloid (ABri) peptide is precipitated as amyloid fibrils in pathological lesions which are characteristic of familial British dementia. Unlike for other amyloidogenic peptides which have been implicated in neurodegenerative disease, for example, Abeta in Alzheimer's disease and alpha synuclein in Parkinson's disease, nothing is yet known as to whether metals mediate the formation of ABri amyloid fibrils. We show herein that a concentration of ABri, which had not previously been shown to spontaneously form amyloid, formed fibrils when incubated for 12 months at 37 degrees C. The additional presence of Al(III), in particular, or Fe(III) increased significantly both the number and the size of the fibrillar amyloid deposits which were very similar in appearance to amyloid described in hippocampal plaques in familial British dementia. Co-incubation of ABri with either Zn(II) or Cu(II) precipitated the peptide but did not result in the formation of amyloid fibrils. PMID- 15541489 TI - A comparative study of the interaction of two structurally analogue ruthenium(II) complexes with DNA. AB - The binding of the ruthenium(II) complexes of [Ru(bpy)2(CAIP)]Cl2 and [Ru(bpy)2(HCIP)]Cl2 (where bpy=2,2'-bipyridine, CAIP=4-carboxyl-imidado[4,5 f][1,10]-phenanthroline, HCIP=3-hydroxyl-4-carboxyl-imidado[4,5-f][1,10] phenanthroline) to calf thymus DNA (ct-DNA) has been investigated with UV-visible and emission spectroscopy, steady-state emission quenching, and viscosity measurements. The experimental results indicate that the two complexes bind to ct DNA through an intercalative mode and [Ru(bpy)2(HCIP)]2+ intercalates into DNA more deeply than [Ru(bpy)2(CAIP)]2+ does. PMID- 15541490 TI - The copper(II) binding properties of the cyclic peptide c(HGHK). AB - The new cyclic tetrapeptide c(HGHK) was synthesised in the solid phase and its complexes with copper(II) were studied in aqueous solution at various pH values by means of potentiometric and spectroscopic methods (UV, EPR, CD). Six mononuclear coordination species were clearly identified within the pH range 3 11. Spectroscopic data strongly suggest sequential formation of N, 2N, 3N and 4N equatorial donor sets around the copper(II) centre from the lowest to the highest pH, involving both imidazole nitrogens and amide nitrogens. A detailed comparison with the copper(II) binding properties of HGHG and Ac-HGHG ligands is also reported. PMID- 15541491 TI - Orally administrated aluminum-maltolate complex enhances oxidative stress in the organs of mice. AB - Recently, aluminum (Al) in drinking water has been proposed to be a risk factor for development of Alzheimer's disease (AD). Because the physiological role of Al in humans is not yet known, we previously examined this role using an experimental animal model. Our results revealed a greater accumulation of Al in the brain, liver, kidney and spleen of mice who received long-term (90 days) administration of an Al complex, aluminum-maltolate (ALM), than in untreated controls. This observation prompted us to examine the degree of injury in the organs of mice in terms of lipid peroxidation evaluated by thiobarbituric acid reactive substances (TBARS) and NOx levels in order to determine the effects of Al accumulation. Six-week-old mice were given drinking water containing AlCl3 or ALM for 120 days. TBARS and NOx levels were found to change depending on the organs and chemical forms of Al. In particular, TBARS and NOx levels in the brain of mice given ALM for 30, 60 and 120 days were significantly increased compared with those of the control group. In addition, nervous degeneration was detected in the brain of the ALM-treated group. These results indicate that the chemical form of Al alters the distribution and oxidative stress in the brain. In addition, we propose a more precise method of determining Al levels in biological systems using neutron activation analysis. When the biological samples are irradiated with a neutron flux, both 27Al and 31P can be counted to 28Al by the reactions, 27Al(n,gamma)28Al and 31P(n,alpha)28Al, respectively. The level of Al in the organs can then be determined by subtraction of the radioactivity due to 28Al originated in 31P from the total radioactivity of the samples. PMID- 15541492 TI - A comparative study of clinically well-characterized human atherosclerotic plaques with histological, chemical, and ultrastructural methods. AB - Atherosclerotic plaques (six cases) with well-documented clinical history were analysed using histology, scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive X-ray (EDX) spectroscopy, infrared spectroscopy (IR), thermogravimetry (TG), and high-resolution synchrotron X-ray diffraction. All samples contained about 60-70 wt% biological carbonated apatite (in dry state) in a nanocrystalline form with particle sizes of about 20 nm. Structurally, there are strong similarities to bone mineral. Ultrastructural investigations documented typical calcospherites, mineralisation processes starting at collagen fibrils and ring-shaped crystalline mineralised structures. There were no significant ultrastructural or chemical differences between the calcifications of individual patients. PMID- 15541493 TI - Complexes of amylose and amylopectins with multivalent metal salts. AB - Metal cations [Cu(II), Fe(III), Mn(II), and Ni(II)] are ligated by amylose as well as potato, and corn amylopectins as proven by electron paramagnetic resonance spectra and conductivity measurements. The hydroxyl groups of polysaccharides are the coordination sites. Isolated starch polysaccharides did not coordinate to metal ions so well as starch did. The resulting polycenter Werner complexes were mainly square planar species. The ligation of the central metal atoms resulted in a variation of the thermal stability, pathway, and rate of thermal decomposition of starch as proven by thermogravimetric (TG, DTG) and scanning differential calorimetric measurements. Frequently, amylose and potato amylopectin willingly formed clathrates in which the water molecules were caged. The mode of the coordination of the hydroxyl groups to the central metal atom controlled the clathrate formation from amylose and in the case of potato amylopectin metal atoms bound to the phosphoric acid moiety formed cage by coordination of the hydroxyl groups to them. Coordination to selected metal salts controls pathway and products of polysaccharide ligand thermolysis. PMID- 15541494 TI - Synthetic analogue approach to metallobleomycins: syntheses, structure and properties of mononuclear and tetranuclear gallium(III) complexes of a ligand that resembles the metal-binding site of bleomycin. AB - As part of our interest into the bioinorganic chemistry of gallium, gallium(III) complexes of the peptide ligand N-(2-(4-imidazolyl)ethyl)pyridine-2-carboxamide (pypepH2) resembling a fragment of the metal-binding domain of bleomycins (BLMs), have been isolated. Reaction of pypepH2 with (Et4N)[GaCl4] and Ga(acac)3 [acac- is the acetylacetonate(-1) ion] affords the mononuclear complex [Ga(pypepH)2]Cl.2H2O (1) and the tetranuclear complex [Ga4(acac)4(pypep)4].4.4H2O (2), respectively. Both complexes were characterized by single-crystal X-ray crystallography, IR spectroscopy and thermal decomposition data. The pypepH- ion in 1 behaves as a N(pyridyl), N(deprotonated amide), N(pyridine-type imidazole) chelating ligand. The doubly deprotonated pypep2- ion in 2 behaves as a N(pyridyl), N(deprotonated amide), N(imidazolate), N'(imidazolate) mu2 ligand and binds to one Ga(III) atom at its pyridyl, amide and one of the imidazolate nitrogens, and to a second metal ion at the other imidazolate nitrogen; a chelating acac- ligand completes six coordination at each Ga(III) centre. The IR data are discussed in terms of the nature of bonding and known structures. The 1H NMR spectrum of 1 suggests that the cation of the complex maintains its integrity in dimethylsulfoxide (DMSO) solution. Complexes 1 and 2 are the first synthetic analogues of metallobleomycins with gallium(III). PMID- 15541495 TI - Complementary inhibition of synoviocyte, smooth muscle cell or mouse lymphoma cell proliferation by a vanadyl curcumin complex compared to curcumin alone. AB - A novel vanadyl curcumin complex (VO(cur)2) has been synthesized and and its physicochemical properties characterized. Biological characterization included in vitro testing for anti-rheumatic activity in synoviocytes, angiogenesis inhibition in smooth muscle cells and anti-cancer potential in mouse lymphoma cells; as well as in vivo testing for hypoglycemic activity by oral gavage in streptozotocin (STZ)-diabetic rats. VO(cur)2 was more effective as an anti-cancer agent, compared to uncomplexed curcumin or vanadyl ion alone, was more than twice as effective as curcumin alone as an anti-arthritic agent, and was more than four times as effective as curcumin alone in inhibiting smooth muscle cell proliferation. In both acute and chronic screening tests, VO(cur)2 was ineffective as an insulin mimetic agent; however, it also proved to be exceptionally non-toxic, with no evidence of negative symptomatology during a month-long treatment period, at doses up to and including 2.0 mmol kg(-1) day( 1). PMID- 15541496 TI - Synthesis, characterization, and cytotoxic activity of copper(II) and platinum(II) complexes of 2-benzoylpyrrole and X-ray structure of bis[2 benzoylpyrrolato(N,O)]copper(II). AB - Copper(II) and platinum(II) complexes of 2-benzoylpyrrole (2-BZPH) were synthesized and characterized with IR, 1H and 13C NMR spectroscopies and coordination geometry with ligands arranged in transoid fashion. The crystal structure of [Cu(II)(2-BZP)2] was determined by X-ray diffraction. Death of complex treated Jurkat cells was measured by flow cytometry. The bis-chelate complexes [Cu(II)(2-BZP)2] and [Pt(II)(2-BZP)2] adopt square-planar coordination geometry with ligands, arranged in transoid fashion. Concentrations of 1-10 microM Platinum(II) complexes reduced cell survival from 100% to 20%, in contrast to the copper(II) complex which caused no cell death at a concentration of 10 microM. While the Pt(II) complexes may have damaged DNA to induce cell death, treatment with the Cu(II) complex did not induce Jurkat cell death. PMID- 15541497 TI - Effects of chronic treatment with sodium tetrachloroaurate(III) in mice and membrane models. AB - Gold is a nonessential element with a variety of applications in medicine. A few gold(I) compounds are used in the clinics for treatment of rheumatoid arthritis and of discoid lupus. Some novel gold(III) compounds are under evaluation as anticancer agents. It is known that gold compounds generally produce toxic effects on the kidneys and characteristic lesions in the brain. However, information concerning the neurotoxicity of gold derivatives in humans as well as in experimental toxicology is rather scarce. For this reason we tried to shed some further light on this aspect of gold neurotoxicity by chronic treatment of mice with sodium tetrachloroaurate(III) in order to observe possible biophysical and morphological alterations that may occur in the brain. Chronic gold treatment resulted in a markedly decreased expression of metallothioneins and of glial fibrillary acidic protein in astrocytes of different brain areas. To examine its effects on cell membranes, interactions of sodium tetrachloroaurate(III) with molecular models were also evaluated. The models consisted in bilayers built-up of classes of phospholipids located in the outer and inner monolayers of biological membranes. Structural perturbation of cell membrane models was observed only at concentrations 10(5) times higher than those detected in the brains of animals after three months' treatment. These results show that toxic effects on animal brain upon treatment with sodium tetrachloroaurate develop with difficulty and may be observed only at high doses. PMID- 15541499 TI - A novel cytotoxic ternary copper(II) complex of 1,10-phenanthroline and L threonine with DNA nuclease activity. AB - A novel ternary copper(II) complex, [Cu(phen)(L-Thr)(H2O)](ClO4) (phen=1,10 phenanthroline, L-Thr=L-threonine), has been synthesized and structurally characterized. The complex crystallized in a triclinic system with space group P1 , a=7.526(15) A, b=11.651(2) A, c=12.127(2) A, alpha=115.41(3) degrees , beta=102.34(3) degrees and gamma=91.33(3) degrees . The copper(II) center is situated in a distorted square-pyramidal geometry. At a concentration of 10(-6) mol L(-1), the complex exhibited potent cytotoxic effects against human leukemia cell line HL-60 and human stomach cancer cell line SGC-7901 with inhibition rates of over 90%, however, less pronounced effects were observed for human liver carcinoma cell line BEL-7402 and human non-small-cell lung cancer cell line A 549. The complex was shown to bind DNA by intercalation and cleave pBR322 DNA in the presence of ascorbate. PMID- 15541498 TI - Vanadium(III) complexes with L-cysteine--stability, speciation and the effect on actin in hepatoma Morris 5123 cells. AB - The complexation processes of vanadium(III) with L-cysteinate and s-methyl-L cysteinate ligands have been studied in aqueous solutions in the pH range 2-7 by the pH-potentiometric, UV-Vis absorption and CD spectroscopy methods. The equilibria model of complex formation, evaluated by SUPERQUAD program, so as careful inspection of spectroscopic data have allowed to determine the speciation and the coordination mode of vanadium(III) ion in the major species present in aqueous solutions. Relatively stable ML2 species of vanadium(III)-L-cysteinate system exists in aqueous solutions above pH 5. It was deduced from spectral data that the coordination sphere of vanadium(III) ion in V(Cys)2 is completed by oxygen, nitrogen and sulfur atoms of two L-cysteinate ligands. Solution of vanadium(III) with L-cysteine (pH approximately 7, L/M=20) was administrated to the culture medium of hepatoma Morris 5123 growing cells. Cytotoxic effect of this solution towards tumor cells was observed. The viability of these cells depended on the complex concentration. It was reduced by 70% at 100 microM of the vanadium species concentration in the culture medium. The death of cancer cells seems to be induced on apoptotic route. The statistically significant increase of total actin level and filamentous to monomeric actin ratios (F/G) were found in the cytoplasm of cells exposed to the vanadium(III)-L-cysteine complex. It was accompanied by the rearrangement of actin cytoskeleton architecture. These factors are important for migration and metastasis formation of the cancer cells. PMID- 15541500 TI - Mimic models of peroxidase--kinetic studies of the catalytic oxidation of hydroquinone by H2O2. AB - The reactions of hydroquinone with hydrogen peroxide catalyzed by transition metal ions Cu2+, Fe2+, Fe3+, Co2+ and Mn2+ were investigated in aqueous solution at 25 degrees C. Two copper (II) complexes (bis(dimethylglyoxime) copper(II) and 5,7,7,12,14,14-hexamethyl-1,4,8,11-tetraazacyclotetradeca-4,11 dienatocopper(II)iodide) were prepared. Their catalytic activities on this oxidation were kinetically investigated in aqueous solution and in cetyltrimethylammonium bromide (CTAB) micellar solution at 25 degrees C. The kinetic equations for micellar catalysis and metallomicellar catalysis were established, respectively. CTAB micelle enhances the reaction rate due to its concentrated and electrostatic effects on substrates and/or intermediate. Metallomicelle exhibits remarkable catalytic activity on this oxidation reaction, which is attributed to the active center and the microenvironment effects. Metallomicelle enhances the rate of reaction by activating hydroquinone anion. The presence of co-ligand of imidazole (or pyridine) remarkably increases the catalytic activity of metal complex in micelle system in contrast to it lowers the activity of the complex in aqueous solution. Metallomicelles could be treated as the mimic models of peroxidase. PMID- 15541502 TI - Preparation and characterization of a (Cu,Zn)-pMMO from Methylococcus capsulatus (Bath). AB - We report the preparation of a (Cu,Zn)-particulate methane monooxygenase (pMMO) in which the bulk of the copper ions of the electron-transfer clusters (E clusters) has been replaced by divalent Zn ions. The Cu and Zn contents in the (Cu,Zn)-pMMO were determined by both inductively coupled plasma mass spectroscopy (ICP-MS) and X-ray absorption K-edge spectroscopy. Further characterization of the (Cu,Zn)-pMMO was provided by pMMO-activity assays as well as low-temperature electron paramagnetic resonance (EPR) spectroscopy following reductive titration and incubation in air or air/propylene mixtures. The pMMO-activity assays indicated that the (Cu,Zn)-pMMO was no longer capable of supporting catalytic turnover of hydrocarbon substrates. However, the EPR studies revealed that the catalytic cluster (C-cluster) copper ions in the (Cu,Zn)-pMMO were still capable of supporting the activation of dioxygen when reduced, and that the 14N superhyperfine features associated with one of the type 2 Cu(II) centers in the hydroxylation C-cluster remained unperturbed. The replacement of the E-cluster copper ions by Zn ions did compromise the ability of the protein to mediate the transfer of reducing equivalents from exogenous reductants to the C-clusters. These observations provide strong support for the electron transfer and catalytic roles for the E-cluster and C-cluster copper ions, respectively. PMID- 15541501 TI - Intramolecular stacking interactions in ternary copper(II) complexes formed by a heteroaromatic amine and 9-[2-(2-phosphonoethoxy)ethyl]adenine, a relative of the antiviral nucleotide analogue 9-[2-(phosphonomethoxy)ethyl]adenine. AB - The stability constants of the mixed-ligand complexes formed between Cu(Arm)2+, where Arm=2,2'-bipyridine (Bpy) or 1,10-phenanthroline (Phen), and the dianions of 9-[2-(2-phosphonoethoxy)ethyl]adenine (PEEA2-) and (2-phosphonoethoxy)ethane (PEE2-), also known as [2-(2-ethoxy)ethyl]phosphonate, were determined by potentiometric pH titrations in aqueous solution (25 degrees C; I=0.1 M, NaNO3). The ternary Cu(Arm)(PEEA) complexes are considerably more stable than the corresponding Cu(Arm)(R-PO3) species, where R-PO3(2-) represents a phosph(on)ate ligand with a group R that is unable to participate in any kind of interaction within the complexes. The increased stability is attributed to intramolecular stack formation in the Cu(Arm)(PEEA) complexes and also, to a smaller extent, to the formation of 6-membered chelates involving the ether oxygen atom present in the -CH2-O-CH2-CH2-PO3(2-) residue of PEEA2-. This latter interaction is separately quantified by studying the ternary Cu(Arm)(PEE) complexes which can form the 6-membered chelates but where no intramolecular ligand-ligand stacking is possible. Application of these results allows a quantitative analysis of the intramolecular equilibria involving three structurally different Cu(Arm)(PEEA) species; e.g., of the Cu(Bpy)(PEEA) system about 11% exist with the metal ion solely coordinated to the phosphonate group, 4% as a 6-membered chelate involving the ether oxygen atom of the -CH2-O-CH2CH2-PO3(2-) residue, and 85% with an intramolecular stack between the adenine moiety of PEEA2- and the aromatic rings of Bpy. In addition, the Cu(Arm)(PEEA) complexes may be protonated, leading to Cu(Arm)(H;PEEA)+ species for which it is concluded that the proton is located at the phosphonate group and that the complexes are mainly formed (50 and 70%) by a stacking adduct between Cu(Arm)2+ and the adenine residue of H(PEEA)-. Finally, the stacking properties of adenosine 5'-monophosphate (AMP2-), of the dianion of 9-[2-(phophonomethoxy)ethyl]adenine (PMEA2-) and of several of its analogues (=PA2-) are compared in their ternary Cu(Arm)(AMP) and Cu(Arm)(PA) systems. Conclusions regarding the antiviral properties of several acyclic nucleoside phosphonates are shortly discussed. PMID- 15541503 TI - Synthesis, characterization, EXAFS investigation and antibacterial activities of new ruthenium(III) complexes containing tetradentate Schiff base. AB - A series of new hexa-coordinated ruthenium(III) complexes of the type [Ru(X)(2 atmp-ba)(EPh3)] (where H2-2-atmp-ba=N,N'-bis(2-aminothiophenol)benzoylacetone; X=Cl or Br; E=P or As) have been prepared by reacting [RuX3(EPh3)3] (where X=Cl or Br; E=P or As) with tetradentate Schiff base ligand (H2-2-atmp-ba) in 1:1 molar ratio. The complexes have been characterized by elemental analyses, Infra red, electronic, electron paramagnetic resonance spectroscopy and cyclic voltammetry. In order to confirm the coordination and structure of the complexes extended X-ray absorption fine structure spectroscopy (EXAFS) studies have been carried out. Based on the above data, an octahedral structure has been confirmed for the complexes. The new complexes were also screened for their antibacterial properties. PMID- 15541504 TI - Identification of copper(II) binding sites in actinomycin D, a cytostatic drug- correlation of coordination with DNA damage. AB - Actinomycin D (AD) is a potent anticancer drug widely applied in therapy, which however exhibits very high toxicity in humans. As the character of donors present in the AD molecule seems to be very favorable for Cu(II) ions, we undertook the coordination study on the Cu(II)-AD system. Potentiometric experiments proved a formation of very stable complexes and with the use of spectroscopic methods the identification of the binding sites was made. The values of potential energy minima, provided by theoretical modeling, confirmed the feasibility of formation of the complexes in water solution. We also demonstrated a significant effect of Cu(II) ions on AD interactions with DNA. The strand-nicking activity was observed. This process could be correlated with the speciation of complex forms. We also found out that in the presence of H2O2, low levels of Cu(II)-AD complexes induce the formation of considerable amounts of linearised plasmid. In consequence, the hypothesis is proposed that the physiologically available cupric ions may participate in the drug-induced toxic effects. PMID- 15541505 TI - Characterization and crystal structure of cadmium(II) halide complexes with amino acids and their derivatives VI. The comparison of crystal structures of cadmium(II) halide complexes with three kinds of piperidine carboxylic acids. AB - Six cadmium(II) halide complexes with dl-piperidine-2-carboxylic acid (DL-Hpipe 2), dl-piperidine-3-carboxylic acid (DL-Hpipe-3), and piperidine-4-carboxylic acid (Hpipe-4), have been prepared and characterized by means of IR and Raman spectra and thermal analysis. The crystal structures of [CdCl2(DL-Hpipe-2)(H2O)], [CdBr2(DL-Hpipe-3)], and [CdCl2(Hpipe-4)] have been determined by X-ray diffraction. These three complexes have one-dimensional polymer structures bridged by halide atoms. The crystal of [CdCl2(DL-Hpipe-2)(H2O)] is orthorhombic with the space group Pca2(1). The cadmium atom is in an octahedral geometry, ligated by a carboxyl oxygen atom, two bridging chlorine atoms, a terminal chlorine atom, a water molecule and a carboxyl oxygen atom of a neighboring molecule. The carboxyl oxygen atoms of DL-Hpipe-2 are coordinated to two cadmium atoms. The unit cell consists of two types of one-dimensional polymer structures: [CdCl2(D-Hpipe-2)(H2O)] and [CdCl2(L-Hpipe-2)(H2O)]. Therefore, it is better to write [CdCl2(DL-Hpipe-2)(H2O)] as [CdCl2(D-Hpipe-2)(H2O)][CdCl2(L-Hpipe-2)(H2O)]. The crystal structure of [CdBr2(DL-Hpipe-3)] is monoclinic with space group P2(1). The cadmium atom is in a distorted octahedral geometry ligated by two carboxyl oxygen atoms and four bridging bromine atoms. This complex consists of either D-Hpipe-3 or L-Hpipe-3. Therefore [CdBr2(DL-Hpipe-3)] is written as [CdBr2(D or L-Hpipe-3)]. The crystal of [CdCl2(Hpipe-4)] is monoclinic with space group P2(1)/n. The structure is similar to that of [CdBr2(D or L-Hpipe-3)]. PMID- 15541506 TI - Inhibition of key cytokines by tetrathiomolybdate in the bleomycin model of pulmonary fibrosis. AB - Tetrathiomolybdate is an anticopper drug with a unique mechanism of action. Tetrathiomolybdate complexes copper to protein and itself, rendering the copper unavailable for cellular uptake. It was originally developed for Wilson's disease, and is now being developed as an antiangiogenic agent for the treatment of cancer. Many angiogenic cytokines require normal levels of copper, and lowered copper levels reduce cytokine signaling while cellular copper requirements are met. Cytokines of fibrosis and inflammation may be similarly copper dependent, since tetrathiomolybdate inhibits bleomycin induced pulmonary inflammation and fibrosis. The basis for this inhibition was evaluated here by examination of tetrathiomolybdate effects on cytokines in lung pathophysiologically important in the bleomycin mouse model of pulmonary damage. Results in mice injected endotracheally with bleomycin confirmed that tetrathiomolybdate therapy was effective in reducing fibrosis. This effect was associated with significant inhibition of bleomycin-induced tumor necrosis factor alpha and transforming growth factor beta expression in lung homogenates. These effects were shown to be independent of one another. This indicates that tetrathiomolybdate therapy can be effective even when fibrosis is at a more chronic stage, wherein inflammatory cytokines are playing a diminishing role. The inhibition of tumor necrosis factor alpha suggests that diseases of tumor necrosis factor alpha overexpression are also potential targets of tetrathiomolybdate therapy. PMID- 15541507 TI - Novel ruthenium complex K2[Ru(dmgly)Cl4].2H2O is toxic to C6 astrocytoma cell line, but not to primary rat astrocytes. AB - A novel class of ruthenium (III) complexes of formulas K[Ru(sar)2Cl2].12H2O and K2[Ru(dmgly)Cl4].2H2O, containing bidentate chelates N-methylglycine (sarcosine, sar) or N,N-dimethylglycine (dmgly) and additional chloro ligands were synthesized. The complexes have been obtained by direct reaction of ruthenium(III) chloride with corresponding bidentate ligand followed by addition of base (KOH). These new complexes were characterized by elemental analysis, IR and electronic absorption spectroscopy. As astrocytomas, the most common of all brain tumors, are still very difficult to treat, we examined the influence of newly synthesized ruthenium-based complexes, as well as the earlier synthesized analogue platinum(IV) complexes [Pt(dmgly)2Cl2], [Pt(sar)2Br2] and [Pt(dmgly)2Br2], on rat astrocytoma C6 cells in vitro. Among these complexes only K2[Ru(dmgly)Cl4].2H2O and [Pt(dmgly)2Br2] markedly inhibited the viability of non confluent C6 cells. Furthermore, only complex K2[Ru(dmgly)Cl4].2H2O was able to reduce viability in confluent C6 cultures. Importantly, this complex was not toxic to primary rat astrocytes or macrophages. Having in mind that appropriate chemotherapy should be effective against tumor cells without harming normal tissues, complex K2[Ru(dmgly)Cl4].2H2O could be a promising agent for developing therapeutics against astrocytomas. PMID- 15541509 TI - Dopamine: the salient issue. AB - There is general agreement that midbrain dopamine neurons play key roles in reward processing. What is more controversial is the role they play in processing salient stimuli that are not rewarding. This controversy has arisen for three main reasons. First, salient sensory stimuli such as tones and lights, which are assumed not to be rewarding, increase dopamine neuron activity. Second, aversive stimuli increase firing in a minority of putative dopamine neurons. Third, dopamine release is increased following aversive stimuli. Consequently, it has been suggested that these midbrain dopamine neurons are activated by all salient stimuli, rather than specifically by rewards. However, reconsideration of these issues, in light of new findings, suggests this controversy can be resolved in favour of reward theories. PMID- 15541508 TI - The expanding role of PSD-95: a new link to addiction. AB - One feature of addiction is the enhanced locomotor response known as behavioral sensitization that occurs with prolonged exposure to psychostimulants. In a recent study, Yao et al. have analyzed four distinct animal models that share this common phenotype. Evaluation of gene expression in the striatum reveals that postsynaptic density (PSD)-95, a synaptic scaffolding protein, is downregulated in each model, suggesting an essential role for this protein in addiction. PMID- 15541510 TI - Cognitive consonance: complex brain functions in the fruit fly and its relatives. AB - The fruit fly, Drosophila melanogaster, has become a model for the study of a growing number of human characteristics because of the power of its genetics. Higher cognitive functions, however, might be assumed to be out of reach for the little fly. But the cumulative history of cognitive studies in insects and some of their arachnid relatives, as well as specific probing of the capabilities of fruit flies, suggests that even in this ethereal realm these creatures have much to contribute. What are the degrees of sophistication in cognitive behavior displayed by these organisms, how have they been demonstrated, and what is their potential for understanding how our own brains work? PMID- 15541511 TI - The Bayesian brain: the role of uncertainty in neural coding and computation. AB - To use sensory information efficiently to make judgments and guide action in the world, the brain must represent and use information about uncertainty in its computations for perception and action. Bayesian methods have proven successful in building computational theories for perception and sensorimotor control, and psychophysics is providing a growing body of evidence that human perceptual computations are "Bayes' optimal". This leads to the "Bayesian coding hypothesis": that the brain represents sensory information probabilistically, in the form of probability distributions. Several computational schemes have recently been proposed for how this might be achieved in populations of neurons. Neurophysiological data on the hypothesis, however, is almost non-existent. A major challenge for neuroscientists is to test these ideas experimentally, and so determine whether and how neurons code information about sensory uncertainty. PMID- 15541512 TI - Neurobiology of dyslexia: a reinterpretation of the data. AB - Theories of developmental dyslexia differ on how to best interpret the great variety of symptoms (linguistic, sensory and motor) observed in dyslexic individuals. One approach views dyslexia as a specific phonological deficit, which sometimes co-occurs with a more general sensorimotor syndrome. This article on the neurobiology of dyslexia shows that neurobiological data are indeed consistent with this view, explaining both how a specific phonological deficit might arise, and why a sensorimotor syndrome should be significantly associated with it. This new conceptualisation of the aetiology of dyslexia could generalize to other neurodevelopmental disorders, and might further explain heterogeneity within each disorder and comorbidity between disorders. PMID- 15541513 TI - Divide et Impera--the midbrain-hindbrain boundary and its organizer. AB - The midbrain-hindbrain organizer (MHO) is a signalling centre that orchestrates development of the mesencephalic and anterior metencephalic primordia. In recent years, details have been revealed about the molecular nature of these signals, their transmission and reception, and the regulatory processes associated with MHO function. This article reviews recent progress in understanding the genetic and molecular components of the MHO, and how they synergize to control brain development. PMID- 15541514 TI - Astrocytic control of glutamatergic activity: astrocytes as stars of the show. AB - It is a major recent finding that astrocytes can influence synaptic activity by release of glutamate, but many other glutamate-mediated activities are also controlled by astrocytes. Even the most obvious neuronal function of glutamate - its release as a transmitter - is regulated by astrocytes; these cells are needed for formation of precursors for glutamate synthesis, for reuptake of released transmitter, and for disposal of excess glutamate. Without astrocytic involvement, normal function of glutamatergic neurons is not possible, as exemplified by almost instantaneous abrogation of normal vision and learning upon inhibition of astrocyte-specific metabolic pathways. In addition, astrocytes are essential for production of the neuroprotectant glutathione, yet they can also contribute to neuronal death during ischemia by maintaining glutamine synthesis, enabling neuronal formation of neurotoxic glutamate. PMID- 15541517 TI - Is the learning of goal-directed displacement effector-independent? AB - The present experiment was designed to investigate whether the learning of goal directed locomotion is effector independent. To answer this question a bilateral transfer of learning paradigm was used. We wanted to find out whether learning can be transferred from a trained effector system to an untrained one. Sixteen participants were asked to proceed through virtual hallways, while walking on a treadmill or handling of a joystick, in order to cross a pair of oscillating doors. Participants received 1050 training trials on the specific effector system before being transferred to the untrained one. Results indicated a clear transfer from handling to walking and only a moderate transfer from walking to handling. This asymmetrical transfer provides partial support in favor of the effector independent hypothesis. Both the theoretical implications of this work and the possible mediating effect of calibration are discussed. PMID- 15541515 TI - Discrete synaptic states define a major mechanism of synapse plasticity. AB - Synapses can change their strength in response to afferent activity, a property that might underlie a variety of neural processes such as learning, network synaptic weighting, synapse formation and pruning. Recent work has shown that synapses change their strength by jumping between discrete mechanistic states, rather than by simply moving up and down in a continuum of efficacy. Coincident with this, studies have provided a framework for understanding the potential mechanistic underpinnings of synaptic plastic states. Synaptic plasticity states not only represent a new and fundamental property of CNS synapses, but also can provide a context for understanding outstanding issues in synaptic function, plasticity and development. PMID- 15541518 TI - A new view on visuomotor channels: the case of the disappearing dynamics. AB - A considerable body of kinematic data supports the proposal that independent visuomotor channels are involved in the control of the transport and grip components of reach and grasp. These channels are seen as having separate perceptual inputs, outputs and internal processing and are thought by some to correspond to independent neuroanatomical pathways. The idea that different groups of muscles and biomechanical structures can be controlled independently is attractive, but this kinematically-inspired hypothesis fails to take into account the complexity of the dynamic relationships and their interactions within the neuromusculoskeletal system. Inertial, viscous, centrifugal, coriolis, gravitational and reflex cross couplings exist between efferent drives to muscles and resulting body movements. Rotation at even a single joint generates a complex set of dynamic reaction forces and requires coordinated activation of many muscles throughout the body to maintain posture and balance. In this theoretical paper we present a new view of independent visuomotor channels in the form of an adaptive neural controller that can compensate for the above interactions and decouple the relationships between efferent drives to muscles and resulting body movements. At the same time, the neural controller renders all the dynamics (linear and nonlinear), other than time delays, of the neuromusculoskeletal system, unobservable in the visuomotor relationships. Using the geometry of nonlinear dynamical systems we show that, providing certain constraints on the structure of time delays within the system are satisfied, there exists a neural controller that can render all the dynamics of the neuromusculoskeletal system (except for time delays) unobservable in the responses. The controller simultaneously decouples all the interactive dynamics so that each of the m independent inputs controls one and only one degree of freedom of the response. This means that each degree of freedom in a multi-joint response can be controlled by an independent component of the visual input, a behaviour that has long been observed in visual tracking experiments. The controller effectively establishes m independent visuomotor channels. However, rather than reflecting separate neuroanatomical pathways, the independent channels result from a neural controller with convergent and divergent connections to compensate for the interactive nonlinear dynamics within the neuromusculoskeletal system. This new view of visuomotor channels has implications for neural control processes involved in the acquisition and adaptability of skilled perceptual-motor behaviour in general, as well as for the design of robotic controllers. PMID- 15541519 TI - Effector dynamics of rhythmic wrist activity and its implications for (modeling) bimanual coordination. AB - To examine the role of the effector dynamics of the wrist in the production of rhythmic motor activity, we estimated the phase shifts between the EMG and the task-related output for a rhythmic isometric torque production task and an oscillatory movement, and found a substantial difference (45-52 degrees) between the two. For both tasks, the relation between EMG and task-related output (torque or displacement) was adequately reproduced with a physiologically motivated musculoskeletal model. The model simulations demonstrated the importance of the contribution of passive structures to the overall dynamics and provided an account for the observed phase shifts in the dynamic task. Additional simulations of the musculoskeletal model with added load suggested that particular changes in the phase relation between EMG and movement may follow largely from the intrinsic muscle dynamics, rather than being the result of adaptations in the neural control of joint stiffness. The implications of these results are discussed in relation to (models of) interlimb coordination in rhythmic tasks. PMID- 15541520 TI - Multiple time scales and subsystem embedding in the learning of juggling. AB - To gain insight into the multiform dynamics and integration of remote yet pertinent subsystems into the performance of complex perceptual-motor skills, we recently conducted a series of longitudinal and cross-sectional experiments on the acquisition of 3-ball cascade juggling in which we measured, next to the ball trajectories, postural sway, eye and head movements and respiration. The aim of the present paper is to review the main results and theoretical implications of these experimental studies for understanding skill acquisition. As regards the evolution of the quality of the juggling itself, we found that only certain aspects of throwing and catching were adjusted, while the goal behavior of sustained juggling (operationalized as the number of consecutive throws) and the degree of frequency and phase locking between the ball trajectories, indexing pattern stability, increased monotonically. The latter three aspects evolved at different rates, reflecting the existence of a temporal hierarchy in learning. Postural sway exhibited initial manifestations of task-specific, possibly mechanically induced, modes of 3:1 and 3:2 frequency locking with the ball trajectories and only few transitions between those modes. Functional stability appeared to be enhanced during practice by minimizing the sway amplitudes rather than by adjusting the sway dynamics itself. Eye and point-of-gaze movements also showed instances of 3:1 and 3:2 frequency locking with the ball trajectories; especially establishing a 3:1 locking (horizontal eye movements) appeared to be important. Expert behavior suggested that extended practice promotes reliance on multiple sources of information, allowing the proficient juggler to switch adaptively between functional organizations involving distinct perceptual systems. No consistent coordination between breathing and juggling was found. It was concluded that multiform dynamics, involving hierarchically ordered time scales, underlie the acquisition of complex skills and that the subsystems subserving realization of the task goal become assembled and embedded in a task- and subsystem-specific manner. PMID- 15541521 TI - Stretch reflexes in the human masticatory muscles: a brief review and a new functional role. AB - Stretch reflexes play a vital role in fine-tuning movements and in automatically maintaining posture. This article briefly reviews the operation of the stretch reflex in the human masticatory system. The conventional approach of stretching muscles in an open-loop manner has yielded much valuable information on the operation of this reflex. In particular, it has revealed that stretching the jaw closing muscles evokes a reflex response with two major components. The short latency reflex is favoured when stretches are brisk, but slower stretches evoke an additional long-latency component. In the hand muscles, the long-latency response is transcortical: in the masticatory muscles, it is not. In addition to its role in servo-control of muscle length during chewing, the stretch reflex in the jaw-closing muscles maintains the vertical position of the mandible during vigorous head movements such as those that occur during running, jumping, hopping and other vigorous whole-body movements in which the head moves briskly up and down. This is an interesting model system in which to investigate stretch reflexes with natural stimuli under unrestrained, physiological conditions. PMID- 15541522 TI - Gait adjustments in response to an obstacle are faster than voluntary reactions. AB - It has been reported that obstacle avoidance reactions during gait have very short latencies. This raises the question whether the cortex can be involved, as it is in voluntary reactions. In this study, latencies of obstacle avoidance (OA) reactions were determined and related to latencies of voluntary stride modifications and simple reaction times (SRT) of hand and foot. Twenty-five healthy young adults participated in this study. While they were walking on the treadmill, an obstacle suddenly fell in front of their left leg. The first reaction to the obstacle was the moment at which the differentiated acceleration curve of the foot deviated from the control signal. Latencies of OA reactions were 122 ms (SD 14 ms) on average. Two very different avoidance reactions (lengthening and shortening of the stride) were noticed, but there was no avoidance strategy effect on OA latencies. OA latencies were significantly shorter as compared to latencies of voluntary stride modifications and simple reaction times of hand and foot. The short OA latencies could not only be explained from the dynamic nature of the task. It is suggested that subcortical pathways might be involved in obstacle avoidance. PMID- 15541523 TI - Force matching errors following eccentric exercise. AB - During eccentric exercise contracting muscles are forcibly lengthened, to act as a brake to control motion of the body. A consequence of eccentric exercise is damage to muscle fibres. It has been reported that following the damage there is disturbance to proprioception, in particular, the senses of force and limb position. Force sense was tested in an isometric force-matching task using the elbow flexor muscles of both arms before and after the muscles in one arm had performed 50 eccentric contractions at a strength of 30% of a maximum voluntary contraction (MVC). The exercise led to an immediate reduction of about 40%, in the force generated during an MVC followed by a slow recovery over the next four days, and to the development of delayed onset muscle soreness (DOMS) lasting about the same time. After the exercise, even though participants believed they were making an accurate match, they made large matching errors, in a direction where the exercised arm developed less force than the unexercised arm. This was true whichever arm was used to generate the reference forces, which were in a range of 5-30% of the reference arm's MVC, with visual feedback of the reference arm's force levels provided to the participant. The errors were correlated with the fall in MVC following the exercise, suggesting that participants were not matching force, but the subjective effort needed to generate the force: the same effort producing less force in a muscle weakened by eccentric exercise. The errors were, however, larger than predicted from the measured reduction in MVC, suggesting that factors other than effort might also be contributing. One factor may be DOMS. To test this idea, force matches were done in the presence of pain, induced in unexercised muscles by injection of hypertonic (5%) saline or by the application of noxious heat to the skin over the muscle. Both procedures led to errors in the same direction as those seen after eccentric exercise. PMID- 15541524 TI - Temporal aspects of passive movement-related corticomotor inhibition. AB - We have previously shown that during rhythmic passive movement of the index finger, the amplitude of the motor evoke potential (MEP) of the first dorsal interosseous muscle (FDI) as the index finger moved through mid-range adduction, is significantly reduced compared to rest [Edwards, D. J., Thickbroom, G. W., Byrnes, M. L., Ghosh, S., & Mastaglia, F. L. (2002). Reduced corticomotor excitability with passive movement: A study using Transcranial Magnetic Stimulation. Human Movement Science 21, 533-540]. In the present study we have investigated the time-course of this phenomenon. We found that MEP amplitude was significantly reduced at the mid-range position in the first cycle of movement (50+/-6% of resting baseline values), and did not vary across subsequent cycles (10 cycles in 50 s), but that MEP amplitude returned to baseline values within 1s of cessation of movement. The results suggest that the pattern of afferent discharge set up by the kinematics of the movement acting at spinal or supraspinal levels underlies the inhibition observed, rather than an effect of central origin or a cumulative effect of ongoing cyclic movement. PMID- 15541525 TI - On the nature of phase attraction in sensorimotor synchronization with interleaved auditory sequences. AB - In a task that requires in-phase synchronization of finger taps with an isochronous sequence of target tones that is interleaved with a sequence of distractor tones at various fixed phase relationships, the taps tend to be attracted to the distractor tones, especially when the distractor tones closely precede the target tones [Repp, B. H. (2003a). Phase attraction in sensorimotor synchronization with auditory sequences: Effects of single and periodic distractors on synchronization accuracy. Journal of Experimental Psychology: Human Perception and Performance, 29, 290-309]. The present research addressed two related questions about this distractor effect: (1) Is it a function of the absolute temporal separation or of the relative phase of the two stimulus sequences? (2) Is it the result of perceptual grouping (integration) of target and distractor tones or of simultaneous attraction to two independent sequences? In three experiments, distractor effects were compared across two different sequence rates. The results suggest that absolute temporal separation, not relative phase, is the critical variable. Experiment 3 also included an anti phase tapping task that addressed the second question directly. The results suggest that the attraction of taps to distractor tones is caused mainly by temporal integration of target and distractor tones within a fixed window of 100 150 ms duration, with the earlier-occurring tone being weighted more strongly than the later-occurring one. PMID- 15541526 TI - Attentional costs of coordinating homologous and non-homologous limbs. AB - This study aimed to examine the attentional demands of coordinating movement patterns across limbs. Eighteen participants performed a circle drawing task involving in-phase and anti-phase coordination modes under homologous, contralateral and ipsilateral limb combinations. Results indicated that: (a) attentional focus further stabilised coordination patterns with a cost at the central level; (b) there was an inverse relationship between stability and probe reaction time (RT) for all coordination patterns, that is the stronger the coupling between the limbs the lower the central cost. Overall, the results support previous research suggesting that attention plays an important role in sustaining coordination pattern stability and that the co-variation between coordination stability and central cost can also be extended to coordination across limbs. PMID- 15541527 TI - The effects of viscous loading of the human forearm flexors on the stability of coordination. AB - This experiment investigated whether the stability of rhythmic unimanual movements is primarily a function of perceptual/spatial orientation or neuro mechanical in nature. Eight participants performed rhythmic flexion and extension movements of the left wrist for 30s at a frequency of 2.25 Hz paced by an auditory metronome. Each participant performed 8 flex-on-the-beat trials and 8 extend-on-the-beat trials in one of two load conditions, loaded and unload. In the loaded condition, a servo-controlled torque motor was used to apply a small viscous load that resisted the flexion phase of the movement only. Both the amplitude and frequency of the movement generated in the loaded and unloaded conditions were statistically equivalent. However, in the loaded condition movements in which participants were required to flex-on-the-beat became less stable (more variable) while extend-on-the-beat movements remained unchanged compared with the unload condition. The small alteration in required muscle force was sufficient to result in reliable changes in movement stability even a situation where the movement kinematics were identical. These findings support the notion that muscular constraints, independent of spatial dependencies, can be sufficiently strong to reliably influence coordination in a simple unimanual task. PMID- 15541528 TI - Attending to the non-preferred hand improves bimanual coordination in children. AB - The effect of attentional focus in bimanual coordination was investigated from a developmental perspective by examining performance of right- and left-handed children, 5-8-years and 9-12-years old, on bimanual reciprocal tapping tasks. Attentional focus was either specified, by asking the children to attend to the preferred or to the non-preferred hand, or unspecified for the execution of the tasks. When attention was oriented to the non-preferred hand we found a reduced movement time and a lower frequency of errors. Performance differences for handedness and age-groups were observed when the children were oriented to attend to the preferred hand or when there was no instruction regarding attention. These differences in performance were eliminated when attention was oriented to the non preferred hand. PMID- 15541529 TI - Development of unimanual versus bimanual task performance in an isometric task. AB - Sixty-three children between 5 and 12 years of age and 15 adults performed a unimanual and a bimanual isometric force task. The performance of the preferred hand in the unimanual task was compared to the performance of the preferred hand in the bimanual task. It was hypothesized that in the bimanual task the absolute error will be higher, there will be more irregularity and the participants will need more time due to the additional effort from the central nervous system, especially with respect to the communication between the hemispheres. Furthermore, in younger children bimanual force variability was expected to be higher due to developmental aspects concerning callosal maturation and attention. It was found that with respect to force generation the preferred hand was not affected by bilateral isometric force generation, but with respect to force regulation it was. The coefficient of variation (CV) of the force was 34% larger in the bimanual task as compared to the unimanual task. For the time to target force, the increase was 28%. With repetition of the trials the CV decreased in the bimanual task, but only in the youngest age group. During development there was no change in absolute error, yet there was a major reduction in force variability in the bimanual task. It is suggested that improvement in interhemispheric communication and in the ability to focus attention plays a role in the decrease in variability with age. PMID- 15541530 TI - Sensory-motor deficits in children with developmental coordination disorder, attention deficit hyperactivity disorder and autistic disorder. AB - Children who have been diagnosed with any one developmental disorder are very likely to meet diagnostic criteria for some other developmental disorder. Although comorbidity has long been acknowledged in childhood disorders, little is understood about the mechanisms that are responsible for the high level of comorbidity. In a series of studies, we have investigated the link between sensory-motor deficits and developmental disorders. Poor sensory-motor integration has long been implicated as a cause of motor problems in developmental disorders such as developmental coordination disorder (DCD), and our recent research has also investigated sensory-motor deficits in children with attention deficit hyperactivity disorder (ADHD) and autistic disorder. Based on a critical examination of relevant literature and some of our recent research findings, we argue that the importance of poor sensory-motor functioning in discriminating children with different disorders has been underestimated. Poor sensory-motor coordination appears to be linked to DCD, but not ADHD. Also, sensory-motor deficits in children with DCD and autistic disorder may provide insight into some of the social difficulties found in these groups of children. This research will increase our understanding of why children with one developmental disorder typically also have problems in other areas. PMID- 15541531 TI - Simplest tasks have greatest dual task interference with balance in brain injured adults. AB - Attention difficulties and poor balance are both common sequel following a brain injury. This study aimed to determine whether brain injured adults had greater difficulty than controls in performing a basic balance task while concurrently completing several different cognitive tasks varying in visuo-spatial attentional load and complexity. Twenty brain injured adults and 20 age-, sex- and education level-matched controls performed a balance-only task (step stance held for 30 s), five cognitive-only tasks (simple and complex non-spatial, visuo-spatial, and a control articulation task), and both together (dual tasks). Brain injured adults showed a greater centre of pressure (COP) excursion and velocity in all conditions than controls. Brain injured adults also demonstrated greater interference with balance when concurrently performing two cognitive tasks than control subjects. These were the control articulation and the simple non-spatial task. It is likely that distractibility during these simple tasks contributed to an increase in COP motion and interference with postural stability in stance. Performing visuo-spatial tasks concurrently with the balance task did not result in any change in COP motion. Dual task interference in this group is thus unlikely to be due to structural interference. Similarly, as the more complex tasks did not uniformly result in increased interference, a reduction in attentional capacity in the brain injured population is unlikely to be the primary cause of dual task interference in this group. PMID- 15541532 TI - Inter-limb coupling in bimanual rhythmic coordination in Parkinson's disease. AB - Recently, it has been shown that rhythmic inter-limb coordination is disturbed in patients with Parkinson's disease (PD). The present study aims to investigate whether this coordination deficit is primarily the result of an impaired coupling, related to hypoactivation of the supplementary motor area (SMA), or primarily the indirect result of an asymmetrical distribution of PD symptoms over the left and right limbs (a peripheral process). Thirty PD patients and 30 matched control participants tapped with the index fingers anti-phase and left and right leading gallop patterns in four visual feedback conditions. Symmetrically affected participants performed significantly worse than asymmetrically affected and control participants in the gallop patterns. This result suggested that the central deficit has a stronger effect on inter-limb coupling in PD than the neuromuscular and biomechanical asymmetry between the limbs. Detailed analysis of inter-tap intervals (variability and correlation) suggested that this deficit leads to a compensatory asymmetrical inter-limb coupling in the primarily right-affected patient group, and under specific circumstances also in the primarily left-affected patient group. The difference in coordination strategy between left- and right-affected patients suggested that pre-morbid hand preference is an important structural constraint on the coupling strategies available to the participants. PMID- 15541533 TI - Impaired modulation of corticospinal excitability following subthreshold rTMS in focal hand dystonia. AB - Previous studies have demonstrated that subthreshold 1 Hz repetitive transcranial magnetic stimulation (rTMS) causes a decrease in corticospinal excitability in neurologically normal subjects. The effects of subthreshold 1 Hz rTMS upon corticospinal excitability and intracortical inhibition in subjects with focal hand dystonia (FHD) is not yet clear. The purpose of this study was to examine the effects of low intensity 1 Hz rTMS upon these variables in control and FHD subjects. We recorded electromyographic (EMG) from the first dorsal interosseous (FDI) muscle of the dominant hands of seven control subjects, and seven affected hands of five FHD subjects. We used single and paired pulse TMS to examine motor evoked potential (MEP) amplitude, short interval intracortical inhibition (ICI) and silent period duration before, during and after 20 min of low intensity 1 Hz rTMS. MEP amplitude decreased significantly over the course of the rTMS in control subjects, but did not change in FHD subjects. Silent period duration was significantly longer in control subjects after rTMS, but there was no change in FHD subjects. There was no significant change in ICI after rTMS in either subject group, despite the rTMS intensity being set to preferentially activate intracortical inhibitory networks. This suggests that low intensity 1 Hz rTMS may have limited application in the normalisation of inhibitory function in FHD. PMID- 15541569 TI - Pituitary cell lines and their endocrine applications. AB - The pituitary gland is an important component of the endocrine system, and together with the hypothalamus, exerts considerable influence over the functions of other endocrine glands. The hypothalamus either positively or negatively regulates hormonal productions in the pituitary through its release of various trophic hormones which act on specific cell types in the pituitary to secrete a variety of pituitary hormones that are important for growth and development, metabolism, reproductive and nervous system functions. The pituitary is divided into three sections-the anterior lobe which constitute the majority of the pituitary mass and is composed primarily of five hormone-producing cell types (thyrotropes, lactotropes, corticotropes, somatotropes and gonadotropes) each secreting thyrotropin, prolactin, ACTH, growth hormone and gonadotropins (FSH and LH) respectively. There is also a sixth cell type in the anterior lobe-the non endocrine, agranular, folliculostellate cells. The intermediate lobe produces melanocyte-stimulating hormone and endorphins, whereas the posterior lobe secretes anti-diuretic hormone (vasopressin) and oxytocin. Representative cell lines of all the six cell types of the anterior pituitary have been established and have provided valuable information on genealogy of the various cell lineages, endocrine feedback control of hormone synthesis and secretions, intrapituitary interactions between the various cell types, as well as the role of specific transcription factors that determine each differentiated cell phenotype. In this review, we will discuss the morphology and function of the cell types that make up the anterior pituitary, and the characteristics of the various functional anterior pituitary cell systems that have been established to be representative of each anterior pituitary cell lineage. PMID- 15541570 TI - Adrenocortical cell lines. AB - The human adrenal cortex is a complex endocrine organ that secretes mineralocorticoids, glucocorticoids and adrenal androgens. These steroids arise from morphologically and biochemically distinct zones of the adrenal gland. Studying secretion of these distinct steroid hormones can make use of cells isolated from the adrenal gland but this requires animal sacrifice and the need for continued isolation for long-term studies. In addition primary cultures of adrenal cells have a limited life-span in culture and the cultured cells are often contaminated by the presence of non-steroidogenic cells. For that reason in vitro cell culture models have several benefits for research on adrenocortical function. Herein we discuss the available adrenocortical cell lines and their uses as model systems for adrenal studies. Focus is placed on the human NCI-H295 and mouse Y-1 adrenal cell lines, which have been used extensively as adrenocortical model systems. These cell lines have proven to be of considerable value in studying the molecular and biochemical mechanisms controlling adrenal steroidogenesis. The current review will discuss the attributes and limitations of the currently available adrenocortical cell lines as models for adrenal studies. PMID- 15541571 TI - Useful cell lines derived from the adrenal medulla. AB - Five approaches for the preparation of adrenal chromaffin cell lines have been developed. Initially, continuous chromaffin lines were derived from spontaneous pheochromocytoma tumors of the medulla, either from murine or human sources, such as the rat PC12 cell line and the human KNA and KAT45 cell lines. Over the last few decades, more sophisticated molecular methods have allowed for induced tumorigenesis and targeted oncogenesis in vivo, where isolation of specific populations of mouse cell lines of endocrine origin have resulted in model cells to examine a variety of regulatory pathways in the chromaffin phenotype. As well, conditional immortalization with retroviral infection of chromaffin precursors has provided homogeneous and expandable chromaffin cells for transplant studies in animal models of pain. This same strategy of immortalization with conditionally expressed oncogenes has been expanded recently to create the first disimmortalizable chromaffin cells, with an excisable oncogenic cassette, as might be envisioned for the creation of human chromaffin cell lines. Eventually, as we increase our understanding of regulating the phenotypic fate of chromaffin cells in vitro, stem or progenitor adrenal medullary cell lines will be derived as an alternative source for expansion and clinical use. PMID- 15541572 TI - Testicular cell lines. AB - The range of in vivo or in vitro immortalized cell lines currently available provides a variety of model systems for studies of normal and pathological cell functions. The cell lines have been derived from spontaneous or experimentally induced tumors, or through in vitro immortalization. The transgenic (TG) techniques provide a powerful approach, allowing the production of in vivo animal models for a variety of diseases, including malignant tumors, through tissue specific expression of oncogenes or other tumor-promoting genes. The TG techniques also enable the production of cell lines with specific characteristics, through insertion of desired genes into specific cell types, which can then be immortalized upon cell culture. The use of temperature sensitive immortalizing genes offers an additional advantage of controlling gene expression, including the proliferation and differentiation of the cells to be immortalized. As regards the male reproductive system, a number of cell lines of testicular somatic cells are currently available. This review covers mainly the immortalized cell lines of testicular Leydig and Sertoli cells, with special reference to murine cell lines for the study of testicular endocrine function and tumorigenesis. These cell lines also provide useful tools to investigate the molecular basis of hormone actions and testicular cell interactions. PMID- 15541573 TI - Ovarian granulosa cell lines. AB - The ovary is a complex endocrine gland responsible for production of sex steroids and is the source of fertilizable ova for reproduction. It also produces various growth factors, transcription factors and cytokines that assist in the complex signaling pathways of folliculogenesis. The ovary possesses two primary steroidogenic cell types. The theca cells (and to a lesser extent, the stroma) are responsible for androgen synthesis, and the granulosa cells are responsible for conversion of androgens to estrogens, as well as progesterone synthesis. These cells undergo a transformation in the luteal phase of the menstrual cycle, converting them from estrogen producing, to predominantly progesterone producing cells. Understanding the molecular mechanisms regulating these cells is essential in understanding the regulation of steroidogenesis and reproduction. Creation of appropriate in vitro cell model systems can provide important tools for the study of ovarian function. This has led to the development of ovarian steroidogenic cell lines in several laboratories. Developing theca cell lines has met with limited success. Conversely, numerous human and animal granulosa cell lines have been developed. This review will discuss the existing granulosa cell lines and their characteristics. PMID- 15541574 TI - Cell lines and primary cell cultures in the study of bone cell biology. AB - Bone is a metabolically active and highly organized tissue consisting of a mineral phase of hydroxyapatite and amorphous calcium phosphate crystals deposited in an organic matrix. Bone has two main functions. It forms a rigid skeleton and has a central role in calcium and phosphate homeostasis. The major cell types of bone are osteoblasts, osteoclasts and chondrocytes. In the laboratory, primary cultures or cell lines established from each of these different cell types provide valuable information about the processes of skeletal development, bone formation and bone resorption, leading ultimately, to the formulation of new forms of treatment for common bone diseases such as osteoporosis. PMID- 15541576 TI - Cell lines derived from pancreatic islets. AB - The islets of Langerhans play a major role in control of metabolic fuel homeostasis. The rapid increase in incidence of diabetes worldwide has spurred renewed interest in islet cell biology. However, gaining a detailed understanding of islet function at a molecular and biochemical level has been complicated by the difficulty and high cost associated with isolation of pancreatic islets. Until recently, islet-derived cell lines have represented sub-optimal surrogates for primary cells for functional studies due to their undifferentiated or unstable phenotypic features. New approaches have resulted in isolation and characterization of rodent insulinoma cell lines that retain many key functional attributes of normal islets and have become useful tools in the study of islet cell biology. PMID- 15541575 TI - Endocrine cell lines from the placenta. AB - Cell-lines derived from human placenta and chorion have been used extensively to model the endocrine functions of human trophoblast. In general terms, the endocrine functions of the primary cells and tissues are at least partially replicated within the cell-lines, suggesting that they may be used as appropriate models. There are, however, two major provisos that compromise this generalisation. Firstly, the endocrine function of placenta represents a complex interaction between cytotrophoblast, syncytiotrophoblast and multiple regulators, so a single cell population digested from the normal environment is unlikely to represent this. Secondly, the characterisation of primary trophoblast populations and of cell-lines is incomplete, complicating the assignment of functions to trophoblast populations. Despite these difficulties, useful information has been obtained from the available cell-lines, regardless of whether they have arisen spontaneously, been transformed in vitro, or derived from cancers in vivo. PMID- 15541577 TI - Radiotherapy for breast cancer in patients undergoing breast reconstruction or augmentation. AB - Due to increasing indications for postmastectomy radiotherapy and a growing demand for breast reconstruction or augmentation, increasing numbers of patients are currently being exposed to both these treatments. In view of the wide range of available techniques for breast reconstruction, either prosthetic or autologous, and their various sequencing in relation to radiotherapy, physicians can be faced with numerous clinical situations requiring comprehensive knowledge of the topic. This review discusses physical, radiobiological and clinical aspects of combining breast reconstruction and radiotherapy. The available data indicate the feasibility of such combinations, although at the expense of increased risk of complications and less satisfactory cosmesis. Of the two methods of breast reconstruction: using autologous tissue or prosthesis, the former seems to provide better cosmesis and a lower risk of complications in conjunction with radiotherapy. To minimize the risk of unfavourable outcome, the techniques and timing of both breast reconstruction and radiotherapy should be given meticulous attention. PMID- 15541578 TI - The impact of parental cancer on children and the family: a review of the literature. AB - OBJECTIVE: Children of cancer patients may go through a distressing time. The aim of this review was to survey present knowledge on the impact of parental cancer on children and the family. DESIGN: Studies published between January 1980 and March 2004 addressing emotional, social, behavioural, cognitive and physical functioning of children of a parent diagnosed with cancer, as well as the association with child, parental and familial variables were reviewed. RESULTS: Fifty-two studies were found. Emotional problems in school-aged children (11 years) were reported in several qualitative studies, but in only one quantitative study. Quantitative and qualitative studies reported anxiety and depression in adolescents (12 years), in particular in adolescent daughters of ill mothers. Quantitative studies generally showed no behavioural and social problems in school-aged children and adolescents. One quantitative study found physical complaints in school-aged children. However, qualitative studies revealed behavioural problems in school-aged children and also described restrictions in cognitive and physical functioning in children of all ages. The most consistent variables related to child functioning appeared to be parental psychological functioning, marital satisfaction and family communication. Intervention studies directed to the needs of children and their families reported positive effects. CONCLUSION: While quantitative studies reported especially disturbed emotional functioning, qualitative studies reported problems in all domains of child functioning. Well-designed studies are needed to gain more insight into the psychosocial functioning of children of cancer patients in order to develop tailored care. PMID- 15541579 TI - Selective oestrogen receptor modulators/new antioestrogens: a clinical perspective. AB - Following tamoxifen, the first selective oestrogen receptor modulator (SERM), a number of other antioestrogens have been developed. The first-generation SERMs exhibit cross-resistance with tamoxifen and have agonist effects on the uterus. Toremifene has equal efficacy to tamoxifen and may be useful as a tamoxifen alternative. Efficacy results for droloxifene and idoxifene were disappointing and their clinical development ceased. Response rates for second-generation SERMs such as raloxifene and arzoxifene are also not high, although raloxifene shows promise in the chemoprevention of breast cancer. Paradoxically, high-dose oestrogens are proving to be effective breast cancer treatment with similar responses to tamoxifen in postmenopausal women with advanced disease, although these drugs are not well tolerated. Fulvestrant is a new type of oestrogen receptor (ER) antagonist with no agonist effects, which binds, blocks and degrades the ER. Fulvestrant produces high response rates compared with the SERMs, is not cross-resistant with SERMs or aromatase inhibitors (AIs) and is equally as effective as the AI anastrozole in the treatment of postmenopausal women with advanced breast cancer who have progressed after prior antioestrogen therapy. Pure antioestrogens such as the ER antagonist fulvestrant provide opportunities for therapeutic sequencing with tamoxifen and AIs and offer exciting possibilities for the future treatment of breast cancer. PMID- 15541580 TI - Laparoscopically assisted colectomy is as safe and effective as open colectomy in people with colon cancer Abstracted from: Nelson H, Sargent D, Wieand HS, et al; for the Clinical Outcomes of Surgical Therapy Study Group. A comparison of laparoscopically assisted and open colectomy for colon cancer. N Engl J Med 2004; 350: 2050-2059. PMID- 15541581 TI - Adjuvant chemotherapy with oxaliplatin, in combination with fluorouracil plus leucovorin prolongs disease-free survival, but causes more adverse events in people with stage II or III colon cancer Abstracted from: Andre T, Boni C, Mounedji-Boudiaf L, et al. Multicenter international study of oxaliplatin/5 fluorouracil/leucovorin in the adjuvant treatment of colon cancer (MOSAIC) investigators. Oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment for colon cancer. N Engl J Med 2004;350:2343-51. PMID- 15541582 TI - Bevacizumab in combination with irinotecan plus fluorouracil plus leucovorin chemotherapy prolongs survival but increases adverse events in people with metastatic colorectal cancer. PMID- 15541583 TI - High-quality evidence in oncology from 18th May to 15th June 2004: a summary. PMID- 15541584 TI - Identifying the cutoff score for the PCL-R scale (psychopathy checklist-revised) in a Brazilian forensic population. AB - This study introduces a Portuguese-language version of psychopathy checklist revised (PCL-R) [Harv. Mental Health Lett. 12 (1995) 4] in the Brazilian penitentiary system. Hare's scale is used extensively in many other countries. In a forensic population sample of 56 male subjects classified as psychopaths and non-psychopaths [Manual for the Hare Psychopathy Checklist-Revised, Multi-Health System, Toronto, 1991], 33 correlated to global personality disorder (GPD) and 23 to partial personality disorder (PPD), respectively, subtypes of antisocial personality disorder [Manual for the Hare Psychopathy Checklist-Revised, Multi Health System, Toronto, 1991]. Subjects were evaluated through psychiatric and neurological examinations, review of judicial records, Rorschach and PCL-R. A control group of 30 subjects without criminological or psychiatric history was also evaluated with the same instruments. PCL-R validation and identification of cutoff score for Psychopathy (GPD group) was assessed through the concurrent use of the Rorschach. PCL-R cutoff score for the Brazilian population was set at 23. Sensitivity was determined at 84.8%, and reliability was high (Kappa index = 0.87). GPD individuals were characterized as clearly psychopathic according to PCL-R criteria while PPD individuals can only be considered mildly psychopathic, with better chance of rehabilitation. PMID- 15541585 TI - Concentrations of toluene in the body killed by an injury to the head shortly after ingesting thinner. AB - An autopsy was conducted on a male showing leather-like skin damage, revealing the cause of death to be an injury to the head. Thinner was found scattered around the scene of death, and stomach and intestine contents smelled strongly of solvent. Toxicological analysis was conducted to determine whether or not the solvent was of a lethal level. Using gas chromatography, peaks of toluene, xylene, and ethylbenzene were detected in the blood and gastric contents. No toluene was detected in the urine, and therefore it was concluded that the decedent died of a severe head injury shortly after solvent ingestion. In the literature, toluene concentrations in blood and lung samples were determined as both fatal and non-fatal but clear differences in the fatality of toluene in solid organ samples, namely, the brain, liver and kidneys were shown. The brain is especially useful in postmortem analysis. In this case, the concentration of toluene in the brain was 20.0 microl/g, which was considered as a non-lethal level. PMID- 15541586 TI - Latex agglutination for bacterial antigens and meningococcus PCR: two useful tools in legal sudden deaths. AB - Bacterial infections are considered to be a major cause of sudden deaths. The recognition of infections caused by Neisseria meningitidis is an essential duty of medicolegal offices due to the risk of secondary cases. Since other microorganisms, such as Haemophilus influenzae and Streptococcus pneumoniae, are also involved in infectious sudden deaths, the identification of the pathogen responsible for death is essential in order to establish a positive diagnosis while also preventing secondary meningococcal cases. However, because of the unreliability of culture methods used for autopsy specimens and the fragile nature of the microorganisms, other techniques were used. In this study, the detection of specific antigens of N. meningitidis (serogroups A, B, C, Y and W135), H. influenzae type b, S. pneumoniae and Group B Streptococcus was undertaken in 40 samples from sudden death cases in legal procedures with a latex agglutination test. In addition, a meningococcus polymerase chain reaction (PCR) assay (ctrA, crgA and siaD genes) was also used as a corroboration method for positive N. meningitidis agglutinations. Eleven cases of sudden death were confirmed to be due to meningococcus while one case was confirmed to have been caused by H. influenzae type b fulminant epiglottitis. Rapid laboratory diagnosis of meningococcal infection allowed contacts management and notification to the health authorities. From the point of view of the authors, forensic diagnosis of unascertained deaths should include latex agglutination and meningococcus PCR when a fulminant infection by N. meningitidis or H. influenzae is suspected as well as in deaths where the cause is unclear. PMID- 15541587 TI - Correlation of methamphetamine results and concentrations between head, axillary, and pubic hair. AB - This study was designed to compare the qualitative results and concentrations of methamphetamine (MA) and its metabolite amphetamine (AP) in head hair and hair collected from different parts of the body (axillae and pubis). Hair from subjects (N = 14) suspected MA users was simultaneously collected. Hair preparation involved washing step, fine cutting, overnight extraction, derivatization by the trifluoroacetic anhydride, and gas chromatography/mass spectrometry (GC/MS) using selective ion monitoring. In this study, we found a good correlation of the qualitative results for MA between head hair and hair on other parts of the body, but there were some differences in concentrations of MA and AP. Namely, the concentrations of MA and AP were higher in axillary and pubic hair than in head hair. PMID- 15541588 TI - Pathogenesis of conjunctival petechiae. AB - Conjunctival petechiae from 15 cases (cause of death: different natural and unnatural) were analysed using confocal laser scanning microscopy (CLSM) in order to visualize the kind of the damage within the vessel wall (diapedesis versus rhexis). The pathomorphological findings with multiple ruptures of vessels appearing to be filled to bursting point define the conjunctival petechiae as a rhexis-haemorrhage. PMID- 15541589 TI - Long persistence of rigor mortis at constant low temperature. AB - We studied the persistence of rigor mortis by using physical manipulation. We tested the mobility of the knee on 146 corpses kept under refrigeration at Torino's city mortuary at a constant temperature of +4 degrees C. We found a persistence of complete rigor lasting for 10 days in all the cadavers we kept under observation; and in one case, rigor lasted for 16 days. Between the 11th and the 17th days, a progressively increasing number of corpses showed a change from complete into partial rigor (characterized by partial bending of the articulation). After the 17th day, all the remaining corpses showed partial rigor and in the two cadavers that were kept under observation "a outrance" we found the absolute resolution of rigor mortis occurred on the 28th day. Our results prove that it is possible to find a persistence of rigor mortis that is much longer than the expected when environmental conditions resemble average outdoor winter temperatures in temperate zones. Therefore, this datum must be considered when a corpse is found in those environmental conditions so that when estimating the time of death, we are not misled by the long persistence of rigor mortis. PMID- 15541590 TI - Genetic analysis of 18 X-linked short tandem repeat markers in Korean population. AB - The population genetic data of 18 X-chromosomal short tandem repeat (STR) markers DXS6807, DXS8378, DXS9895, DXS9902, DXS6810, DXS7132, DXS981, DXS6800, DXS9898, DXS6789, DXS101, DXS6797, GATA172D05, GATA165B12, HPRTB, GATA31E08, DXS8377, and DXS7423 were analyzed in samples of unrelated 220 males and 181 females from Korean population. The exact test for genotype distribution of the markers showed no significant deviation from the Hardy-Weinberg equilibrium. Allele frequencies between male and female samples were not significantly different in all examined markers. All examined males and females showed different hemizygotic haplotype and combined genotypes, respectively. Four cases of mutation were found in GATA172D05, GATA31E08, DXS7132, and HPRTB from the analysis of 95 father-child mother trios. Details of X chromosomal STRs in Koreans would be useful in paternity tests and forensic purposes as well as whole X-chromosomal mapping studies. PMID- 15541591 TI - The effect of the ageing of crime scene objects on the results of scent identification line-ups using trained dogs. AB - In a scent identification line-up, a trained dog matches the scent trace left by a perpetrator at the crime scene to the odour of a suspect in a line-up of different odours. The procedures are strictly defined and the results are routinely used by the police and as evidence in court in a number of European countries. This paper describes the effect of ageing of the odour trace collected at the crime scene on the performance of the dogs in recognising the perpetrator in a line-up. The results show that whilst the dogs perform faultlessly in matching odours collected on the same day, the results drop to a lower level and become more variable in the period studied (2 weeks to 6 months). However, the results do not show a systematic decrease in performance. A possible explanation is the development of a steady state in the glass jars containing the perpetrator odour trace after initial differential evaporation of components of the residue or break down of unsaturated components into saturated ones. Prevention of this initial change may prevent the drop in performance observed in this study, thus increasing the reliability of these scent identifications. PMID- 15541592 TI - The evaluation of evidence in the forensic investigation of fire incidents (Part I): an approach using Bayesian networks. AB - The forensic investigation of the origin and cause of a fire incident is a particularly demanding area of expertise. As the available evidence is often incomplete or vague, uncertainty is a key element. The present study is an attempt to approach this through the use of Bayesian networks, which have been found useful in assisting human reasoning in a variety of disciplines in which uncertainty plays a central role. The present paper describes the construction of a Bayesian network (BN) and its use for drawing inferences about propositions of interest, based upon a single, possibly non replicable item of evidence: detected residual quantities of a flammable liquid in fire debris. PMID- 15541593 TI - The evaluation of evidence in the forensic investigation of fire incidents. Part II. Practical examples of the use of Bayesian networks. AB - This paper extends a previous discussion of the use of Bayesian networks for evaluating evidence in the forensic investigation of fire incidents. Bayesian networks are proposed for two casework examples and the practical implications studied in detail. Such networks were found to provide precious support in addressing some of the wide range of issues that affect the coherent evaluation of evidence. PMID- 15541594 TI - The transfer and persistence of petrol on car carpets. AB - The significance of the presence of petrol in motor vehicle fires has often been challenged due to the possibility of a natural occurrence of petrol residues inside the vehicle. Transfer and persistence studies were undertaken to investigate the potential transfer and persistence of petrol onto vehicle carpets through the 'normal' usage of motor vehicles. The results of the transfer study indicate that petrol may be transferred from the external environment in sufficient quantities via the shoes of drivers or passengers to be detected after a 24 h period, but not after 1 week. Low levels of petrol were detectable after 24 h on all carpet mats where the initial volume was 500 microL or more. The level of evaporation of the petrol detected increased with corresponding increases in the time period between transfer and analysis. The results of the persistence study indicate that small volumes of petrol (less than 100 microL) are unlikely to be detected on carpet after a 24 h period, and volumes of less than 1000 microL are unlikely to be detected on acoustic padding after this time period. Larger volumes may be detected after this period, but will generally not be detectable on either carpet or acoustic padding after 4 weeks. In each case, the petrol that is detected exhibits a chromatographic profile of greater than 60% evaporated petrol. These results demonstrate the significance of finding a large volume of fresh or slightly evaporated petrol on car carpet. PMID- 15541595 TI - Use of SPE and LC/TIS/MS/MS for rapid detection and quantitation of ketamine and its metabolite, norketamine, in urine. AB - Ketamine (K) has become more and more popular for drug abuse in recent years. A lot of pre-treatment work such as extraction and derivatizing increase difficulties in the tests for ketamine in biological specimens. A rapid method to detect and quantitate ketamine and its metabolite norketamine in urine used deuterated dilution followed by solid phase extraction and liquid chromatography/TurboIonSpray/tandem mass spectrometry (LC/TIS/MS/MS) is described. Control recovery for both low and high concentrations can reach to 90%. Ten ketamine positive urines were examinated by this method. Concentrations ranged from 114 to 2925 ng/mL and from 453 to 9805 ng/mL for norketamine. The method was sensitive, specific, accurate and provided easy operation to detect and quantitate ketamine and its metabolites in urine. PMID- 15541596 TI - Genetic polymorphisms of 15 STR loci in Han population from Shaanxi (NW China). AB - The genetic polymorphisms of 15 STR loci, namely D3S1358, D5S818, D7S820, D13S317, vWA, FGA, TH01, TPOX, CSF1PO, D16S539, D8S1179, D21S11, D18S51, D19S433 and D2S1338, were studied in 203 unrelated Han population from Shaanxi province using AmpF/STR Identifiler kit. PMID- 15541597 TI - Analysis of STR polymorphisms in the Chao Shan population in South China. AB - Chao Shao area is a littoral under the jurisdiction of Guangdong province, abutting on Fujian. Historically, the area was relatively isolated from other parts of China until 1990s, when it started to take a small number of immigrants from other regions. People residing in this area speak in unique dialect and have distinct lifestyle. Allele frequencies for the 15 short tandem repeats (STR) loci included in the AmpFLSTR Identifiler kit were obtained from a sample of 144 unrelated Chinese born and living in the Chao Shan area, South China. PMID- 15541598 TI - Genetic variation of 15 STR loci (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, D7S820, D16S539, D2S1338, D8S1179, D21S11, D18S51, and D19S433) in populations of north and central Poland. AB - Allele frequencies for 15 short tandem repeats (STRs) included in AmpFlSTR Profiler and AmpFlSTR SGM Plus kits were obtained from populations of Pomorze Gdanskie, Wielkopolska, Kujawy, Pomorze Zachodnie, Mazury and Mazowsze regions of Poland. PMID- 15541599 TI - Data for 15 autosomal STR markers (Powerplex 16 System) from two Tunisian populations: Kesra (Berber) and Zriba (Arab). AB - Allele frequencies, together with some parameters of forensic interest, for 15 STRs included in the Powerplex 16 System (CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, D21S11, FGA, Penta D, Penta E, TH01, TPO and VWA) were estimated from two samples of unrelated individuals from Tunisia, of different ethnicity: Kesra (Berber) and Zriba (Arab). No deviations from Hardy Weinberg equilibrium were observed after Bonferroni's correction for the number of loci analysed. Comparative analyses between our population data and other North African databases showed that significant differences were concentrated on loci with lowest values of diversity (mainly CSF1PO and D13S317), irrespective of ethnicity and geographic location. PMID- 15541600 TI - Bartonella quintana-induced apoptosis inhibition of human endothelial cells is associated with p38 and SAPK/JNK modulation and with stimulation of mitosis. AB - Previous studies demonstrated that live Bartonella quintana often induces angioproliferative lesions in humans. It modulates endothelial cell apoptotic and inflammatory patterns, thus inducing a very early overexpression of caspase 8 and Apaf-1 and increasing mRNA production of TNF-alpha, interleukin-8, and E selectin. However, starting at 10 hours postinfection, the bacteria provoke antiapoptotic effects that induce an increase of bcl-2 gene transcription. To gain further insight into the cellular mechanisms that regulate apoptosis, survival and proliferation, we studied the modulation of mitogen-activated protein kinase (MAPK) and the activation state of cdc2 kinase, which regulates progression into mitosis. Confocal microscopy findings indicated a maximum rate of Bartonella entry into host cells between postinfection hours 6 and 10. Live bacteria caused substantially higher apoptosis of human umbilical vein endothelial cells-cryopreserved (HUVEC-C) than heat- and trypsin-inactivated microorganisms. During the first 6 hours postinfection, B. quintana triggered a peak of apoptosis, induced activation of p38 MAPK and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), with bacterial clusters appearing at the cellular surface of the HUVEC-C. However, at 8 to 24 hours postinfection, B. quintana was internalized and inhibited proapoptotic signals such as p38 MAPK and SAPK/JNK while inducing antiapoptotic signals. Indeed, expression of the bcl-2 gene and the increase of the bcl-2 kinase active form was concomitant to activation of mitosis, as shown by cdc2 protein activation. These data thus suggest that mechanisms that induce mitotic activity and inhibit apoptotic signals may contribute to the ability of B. quintana to cause vascular proliferation. PMID- 15541601 TI - Differential quantitative blood cultures for the diagnosis of catheter-related bloodstream infections associated with short- and long-term catheters: a prospective study. AB - We prospectively evaluated the value of two levels of differential quantitative blood culture (DQBC) ratio (> or =2:1 or > or =5:1) in diagnosing catheter related bloodstream infections (CRBSIs) in patients with malignancy that have short-term and long-term central venous catheters (CVCs) (<30 and > or =30 days of placement). Diagnosis of CRBSIs was based on results of semiquantitative cultures of removed catheters. For short-term CVCs a 5:1 or greater DQBC ratio had an 18% sensitivity, 67% specificity, 18% positive predictive value (PPV) and 67% negative predictive value (NPV), whereas a 2:1 or greater cut-off point for the DQBC was associated with a 45% sensitivity, 48% specificity, 26% PPV, and 68% NPV for the diagnosis of CRBSIs. For long-term CVCs, a cut-off point of 5:1 or greater of the DQBC had an 81% sensitivity, 40% specificity, 36% PPV, and 83% NPV for the diagnosis of CRBSIa, whereas a 2:1 or greater cut-off point had a 90% sensitivity, 30% specificity, 35% PPV, and 88% NPV for the diagnosis of CRBSIs. In conclusion, DQBCs are not diagnostic of CRBSIs for short-term CVCs. In long term CVCs, DQBCs at 2:1 or greater or 5:1 or greater are sensitive but associated with low specificity and positive predictive value. PMID- 15541602 TI - Enzyme immunoassay and immunoblotting analysis of Helicobacter pylori infection in Turkish asymptomatic subjects. AB - Approximately half of the world population is infected with Helicobacter pylori, particularly in developing countries. The aims of the study were to detect H. pylori infection in asymptomatic Turkish subjects, correlate the infection with the associated risk factors, and to evaluate the cytotoxin-associated gene (CagA) status and other H. pylori antigens. Three hundred nine asymptomatic subjects (124 female) 1-82 years of age (average: 31 years) were serologically tested by enzyme immunoassay and immunoblotting. The enzyme immunoassay detected IgG anti H. pylori antibodies in sera of 216 (70%) out of 309 subjects, 132 (61%) male. Infection rates of 42% in subjects <10 years of age, 55% in 10-19 years, 66% in 20-29 years, 78% in 30-39 years, 79% in 40-49 years, 91% in 50-59 years, 100% in 60-69 years, and 80% in those >70 years of age were detected. Subjects >45 years of age had significantly higher antibody responses, odds ratio = 0.16 (95% confidence interval: 0.07-0.37), than those <45 years. H. pylori infection was significantly higher in married subjects, odds ratio = 0.38 (95% confidence interval: 0.20-0.73), and those with low socioeconomic status. No correlation between gender, education, smoking, and nonsteroidal anti-inflammatory drug intake and infection was detected. Immunoblots revealed antibodies to CagA in 58 (83%) of 70 samples tested. H. pylori infection is prevalent in the asymptomatic Turkish subjects. Marital and socioeconomic status was significantly associated with the acquisition of H. pylori. Antibodies to CagA antigen were highly prevalent in these subjects. PMID- 15541603 TI - Tolerance to amphotericin B in clinical isolates of Candida tropicalis. AB - A broth microdilution method was used for testing amphotericin B against 33 clinical isolates of Candida tropicalis. All isolates were in vitro susceptible to the polyene (MIC [minimal inhibitory concentration] < or = 1.0 microg/mL). However, when the isolates were cultured in a medium containing amphotericin B at a concentration of 1.5 microg/mL, a wide interstrain variation of growth rate was observed. Five isolates (15%) proved to be highly tolerant to the drug and grew at a frequency ranging from 1 x 10(-1) to 2 x 10(-2). Twenty-three isolates (70%) grew at a frequency ranging from 1 x 10(-5) to 1 x 10(-8). The remaining five isolates (15%) failed to grow in drug-containing medium. In general, this growth variation was not associated with amphotericin B MICs displayed by the single isolates. In addition, the strains grown in drug-containing medium did not represent amphotericin B-resistant mutants, as shown by the maintenance of MICs similar to those of their respective parent isolates. Killing experiments conducted in selected isolates confirmed a variation of fungicidal activity of amphotericin B. To see whether this phenomenon was associated with a variation of amphotericin B response in vivo, we established an experimental model of systemic murine candidiasis in CD1 mice by intravenous injection of cells belonging to Candida tropicalis 3147 (growth rate at a frequency of 1 x 10(-1) in amphotericin B medium) and Candida tropicalis 4055 (no growth). Low (0.3 mg/kg/day) and high (1 mg/kg/day) doses of amphotericin B were both effective at reducing the fungal burdens in the kidneys of mice infected with either strain (p, 0.01 to 0.02). However, whereas the burden of mice infected with isolate 3147 and treated with the polyene at 0.3 mg/kg/day was reduced by 1.2 +/- 0.25 (mean +/- standard deviation) log10 cfu/g compared to untreated mice, the same dosing regimen yielded a burden reduction of 2.6 +/- 0.07 log10 cfu/g in mice infected with isolate 4055 (p < 0.001). Similarly, amphotericin B at 1 mg/kg/day yielded a burden reduction of 1.8 +/- 0.20 vs. 2.5 +/- 0.30 log10 cfu/g in mice infected with isolates 3147 and 4055, respectively (p < 0.001). Our data revealed a variable pattern of tolerance to amphotericin B among isolates of Candida tropicalis and showed that this phenomenon might influence the rate of organ clearance during therapy. PMID- 15541604 TI - In vitro fluconazole susceptibility of 1565 clinical isolates of Candida species evaluated by the disk diffusion method performed using NCCLS M44-A guidelines. AB - We determined the in vitro activity of fluconazole against 1565 clinical Candida spp. isolates collected from different specimens of non-AIDS outpatients and inpatients in 3 different regions of Italy. Susceptibility testing was performed by agar disk diffusion using the NCCLS document M44-A guidelines. Candida albicans was the most frequently isolated yeast (68%) followed by C. glabrata (15%), C. tropicalis (5%), C. parapsilosis (5%), and C. krusei (5%). Other yeasts represented 4% of all isolates. Of the 1565 isolates tested, 1449 (92.6%) were susceptible (S) to fluconazole, 43 (2.7%) were susceptible dose-dependent (S-DD) and 73 (4.7%) were resistant (R). Almost all (98.2%) of the C. albicans isolates were classified as S or S-DD. Despite its widespread use, fluconazole displayed good activity against the isolates we tested, and the disk diffusion method was confirmed as a reliable approach to the evaluation of in vitro susceptibility of yeasts to this antimycotic agent. PMID- 15541605 TI - Real-time PCR for detection of Trypanosoma brucei in human blood samples. AB - We have developed a real-time PCR assay for detection of Trypanosoma brucei DNA in human blood samples. The PCR was conducted with newly designed primers targeting the 177-bp repeat satellite DNA in T. brucei and with Sybr Green to monitor the amplicon accumulation. DNA purification using Chelex 100 resin was performed on blood samples collected on Whatman FTA cards and was shown to be a simple and quantitative method as revealed by real-time PCR. The detection limit of the assay was 100 trypanosomes per mL blood, corresponding to an analytical sensitivity of 0.1 genome equivalents. Trypanosome DNA was detected in all blood samples from sleeping sickness patients and, furthermore, the identity of the amplicon was confirmed in all assays by dissociation analysis. Although template DNA from blood samples was amplified with significantly lower efficiency than genomic DNA, similar efficiency between all assays ensured quantitative results. No amplicon product was obtained with samples from uninfected individuals. The results indicate that the real-time PCR assay described is a rapid and sensitive method suitable for the detection of T. brucei in human blood samples in routine clinical laboratory practice. PMID- 15541606 TI - Antimicrobial activity of daptomycin against multidrug-resistant Gram-positive strains collected worldwide. AB - Daptomycin is a cyclic lipopeptide recently released for clinical use in the treatment of serious Gram-positive infections in hospitalized patients. We evaluated the in vitro activity of daptomycin tested against recently isolated multidrug-resistant Gram-positive clinical strains. A total of 386 isolates were selected from a large collection of strains from more than 70 centers located in Europe, North America, and South America. The strains were tested by reference broth microdilution methods in Mueller-Hinton broth with 50 mg/L Ca++ against daptomycin. Daptomycin was the most potent compound tested against penicillin resistant Streptococcus pneumoniae with MIC50/90 values at < or =0.12 and 0.25 microg/mL, respectively. Daptomycin was also highly active against vancomycin resistant enterococci and staphylococcal strains with various resistance patterns. Enterococcus faecium showed higher daptomycin MIC values (MIC90, 4 microg/mL) when compared to E. faecalis (MIC90, 1 microg/mL). In summary, resistance to vancomycin, teicoplanin, quinupristin/dalfopristin, or penicillin among the Gram-positive isolates did not adversely influence daptomycin activity. Daptomycin showed a significant potency and spectrum against Gram-positive species, including multidrug-resistant strains, and may represent a reasonable therapeutic option for infections caused by these important pathogens. PMID- 15541607 TI - Effect of levofloxacin on the viability of intracellular Chlamydia pneumoniae and modulation of proinflammatory cytokine production by human monocytes. AB - Although antibiotics are known to affect the intracellular growth of Chlamydia pneumoniae in acute infections, their efficacy in therapy for chronic infections, including atherosclerosis, remains debatable. Human monocyte-derived macrophages (MDM) obtained from monocytes of healthy donors were infected with C. pneumoniae AR-39 and treated with levofloxacin (8 microg/mL) immediately after infection (0 hours) or 24 hours after infection. Levofloxacin treatment at 24 hours, but not at 0 hours, resulted in a significant decrease in the number of C. pneumoniae inclusions within the MDM (p < 0.05). Also decreased were concentrations of proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, IL-6, and IL-8 in the extracellular medium (p < 0.01). Viable counts in titrations remained similar to those in untreated controls. In summary, levofloxacin administered to MDM at serum-attainable levels 24 hours after C. pneumoniae infection significantly decreased inclusion counts and proinflammatory cytokine production, but did not eliminate the C. pneumoniae infection. PMID- 15541608 TI - Bactericidal activity of garenoxacin tested by kill-curve methodology against wild type and QRDR mutant strains of Streptococcus pneumoniae. AB - Kill-curve bactericidal assays over 24 hours were determined for garenoxacin, a novel des-F(6) quinolone, and levofloxacin at achievable serum drug concentrations ranging from 2 to 8 microg/mL. Tested strains included 4 wild type and 2 target quinolone resistance-determining region (QRDR) mutant Streptococcus pneumoniae. Garenoxacin was 16-fold to 32-fold more active than levofloxacin, and mutant strains (3 QRDR sites) had garenoxacin minimum inhibitory concentration (MIC) values at 1 microg/mL (levofloxacin MIC >32 microg/mL, resistant), but equal to the levofloxacin potency against wild type strains. Garenoxacin killed mutant pneumococci with comparable rapidity, as did levofloxacin versus strains without target alterations. Garenoxacin appears to be a widely usable and highly active agent against S. pneumoniae resistant to other quinolones such as levofloxacin. PMID- 15541609 TI - Evaluation of API ID 32C and VITEK-2 to identify Candida dubliniensis. AB - We have compared two diagnostic systems, API-ID32C (bioMerieux) and VITEK-2ID-YST (bioMerieux), in their ability to diagnose 50 clinical isolates of Candida dubliniensis. API identified 48 isolates and VITEK-2 identified 33 of the 50 isolates. Lactic acid assimilation showed highly dispersed results with API, being of special importance, since this test is one of the four that makes it possible to differentiate C. dubliniensis from C. albicans. The detection of one enzyme, Phosphate-4MU, was always positive with VITEK-2 when it should have been negative. Overall, API ID 32C obtained better results than VITEK-2. However, the latter is simpler to use and has a greater database. PMID- 15541610 TI - Improvement of a latex agglutination test for the evaluation of oxacillin resistance in coagulase-negative staphylococci. AB - The "Slidex MRSA Detection" test (Denka Seiken, Japan) is a latex agglutination assay able to detect PBP2a. We evaluated its ability to differentiate mecA positive from mecA-negative coagulase-negative staphylococci. We included 100 coagulase-negative staphylococci clinical isolates belonging to 9 species, 54 mecA positive and 46 mecA negative, as characterized by PCR. The specificity achieved using the manufacturer's instructions was 100%, but the sensitivity was only 57%. To increase sensitivity, we introduced modifications into the standard protocol. Using either large inocula or oxacillin induction before test performance, we achieved 100% sensitivity. PMID- 15541612 TI - The importance of internal quality controls in the molecular detection of bacteria and fungi in blood culture milieu. PMID- 15541615 TI - Whole-patient assessment, goal planning, and inflection points: their role in achieving quality end-of-life care. AB - This article provides a framework for performing whole-patient assessment and goal planning. These clinical tasks involve a multidisciplinary, multidimensional, patient-centered approach to care and a deep appreciation for the complex interplay between the physical, psychological, social, and spiritual aspects of the human experience of dying. This article stresses the iterative nature of whole-patient assessment and goal planning, both of which should be conducted at certain important junctures in a patient's progression to manage effectively the evolving challenges faced by terminally ill persons and their families. This article also provides suggestions on successfully managing the communication challenges in caring for patients near the end of life and their family. PMID- 15541616 TI - Symptom management in the older adult. AB - Palliative care begins at the time of diagnosis of a life-threatening illness and continues beyond the time of death. Defined in the broadest sense, the goal of palliative care is to provide aggressive symptom management and address the psychological and spiritual needs of the patient and the family. This article reviews the management of some symptoms commonly observed in older patients, highlighting treatment considerations specific to the older population. Ultimately the approach to symptoms must be individualized, and treatment decisions must reflect the patient's goals of care. Although symptom management in older patients may be challenging, it is possible to provide care that significantly enhances quality of life throughout the course of illness. PMID- 15541617 TI - Last hours of living. AB - The last hours of living can be one of the most important times in the life of any patient and his/her family. With appropriate preparation and careful management of the process by skilled clinicians, dying and death can be a comfortable and even rewarding experience for everyone involved. After death, careful attention to the grief of survivors can help them cope with their loss and rebuild their lives. PMID- 15541619 TI - Family caregivers in palliative care. AB - This article defines caregiving broadly to include the informal (ie, unpaid) care provided by family members that goes beyond customary and normative social support provided in social relationships. Family caregivers in particular play significant roles in the care of elders with advanced chronic disease and in the context of palliative and end-of-life care. These caregivers typically are involved in critical medical decisions, provide vital assistance with activities of daily living, and carry out most nonpharmacologic and pharmacologic treatment recommendations for community-based elders. This article describes family caregivers and the costs of care and introduces the stress process model to highlight essential caregiving experiences and needs in the context of palliative care. Common interventions used to support caregivers are highlighted, and the bereavement experience among caregivers is discussed. PMID- 15541620 TI - Spirituality, religion, and healing in palliative care. AB - In end-of-life care, attending to spiritual needs ensures that a dying patient has the opportunity to find meaning in the midst of suffering and to have the opportunity for love, compassion, and partnership in their final journey. This article summarizes some of the beliefs and traditions from Judaism, Islam, and Christianity that affect people as they face their own dying and mortality. People who do not participate in any formal religion also have a drive to find meaning in the midst of suffering and dying. They may find this in personal ways. This article presents some practical tools to help clinicians address and respect spiritual and religious issues of patients. It is crucial that our culture and our systems of care for the dying include a spiritual approach so that dying can be meaningful and even filled with hope. PMID- 15541622 TI - Hospice and palliative care in nursing homes. AB - This article reviews the challenges inherent in providing high-quality palliative care to dying nursing home residents and summarizes the efforts to address these challenges. It is suggested that a stronger physician presence and oversight of physicians knowledgeable in palliative care in nursing homes are needed to improve the quality of end-of-life care in nursing homes. PMID- 15541623 TI - Palliative care in hospitals. AB - Hospital-based palliative care teams have evolved as a natural outgrowth of the modern hospice movement. This article examines why these hospital-based palliative care programs have proliferated, how they typically function, and what data exist as to their effectiveness. Crucial steps necessary for the design and implementation of a successful hospital-based palliative care service also are reviewed. PMID- 15541624 TI - Hospice care at the end of life. AB - Hospice care in the United States has evolved from a movement and philosophy to a new medical specialty that addresses sources of suffering at many levels. Hospice interdisciplinary teams use Maslow's hierarchy of human need to integrate the multiple domains that influence patients' well-being and assists in the development of treatment plans to prevent or alleviate suffering. Contributing to the effectiveness of this care is the Medicare Hospice Benefit, which since 1983 has served as a model and a reimbursement mechanism that has encouraged proliferation of hospices to deliver care in homes, hospitals, and long-term care facilities. The whole-person approach of hospice care may benefit all patients and can be integrated into all medical management. PMID- 15541625 TI - Financing palliative care. AB - Palliative care services are financed by a combination of revenue from a variety of sources: hospital, nursing home, hospice, and physician. Cost avoidance is calculated based on the patterns of care of the patients seen compared with control patients. Most programs also use grants and philanthropy as part of their "mix." PMID- 15541626 TI - PACE and hospice: two models of palliative care on the verge of collaboration. AB - This article examines the managed long-term care program PACE (Program for All inclusive Care of the Elderly) and traditional hospice as vehicles to provide quality palliative care. The missions, origins, and structures of the two programs are compared. Options for collaboration between these two models of care are explored. Local communities that share both models benefit significantly when these two models negotiate a successful collaboration. PMID- 15541627 TI - Home care of the frail elderly. AB - Statistics indicate that as the population ages, there will be an increased need for home care. Despite this increased need and the legal, clinical, and ethical obligations for physicians to participate, little training is available to physicians in the area of home care. This article provides physicians with an understanding of home care and how to incorporate it into clinical practice. The goals of this article are to (1) explain what home care is and what services are available, (2) teach physicians how to access these services and oversee their implementation, and (3) show how physicians can be reimbursed for this increasingly important part of their practice. PMID- 15541628 TI - Measurement of mood in adolescents with intellectual disability. AB - To date, there has been limited research into mood responses among adolescents with intellectual disability. One reason for this is the absence of a reliable and valid measure for the assessment of mood among this population. The present study evaluated such a measure among a sample of 135 adolescents with mild intellectual disability. Results supported the factorial validity of a 12-item derivative of the Profile of Mood States, referred to as the Intellectual Disability Mood Scale (IDMS). Convergent and divergent validity was also supported. Overall, the IDMS showed encouraging psychometric characteristics as a measure of mood among adolescents with intellectual disability. It is hoped that the results of this study will stimulate further research to expand our knowledge of mood responses among this population. PMID- 15541629 TI - Psychotropic medication in adults with mental retardation: prevalence, and prescription practices. AB - People with mental retardation comprise an overmedicated population. Studies the last 20 years or so indicate that nearly 50% of people with mental retardation receiving care have been using psychotropic medication, in the treatment of psychiatric disorders and/or problem behaviours. The recent years guidelines for prescription of psychotropics have been developed, emphasizing, e.g. the presence of a diagnosis, consideration of alternatives, and evaluation. In spite of all this, we found that 37% of people with mental retardation in one Norwegian county were using psychotropics, mostly neuroleptics. Moreover, prescriptions frequently violated current guidelines, especially when conducted by general practitioners. For example, a lot of prescriptions had not been indicated by a diagnosis, alternatives to medications had rarely been explored, and evaluation of effects and side effects were exceptions. Psychiatrists complied more with current guidelines. Implications of the findings are discussed. PMID- 15541630 TI - Pharmacological management of challenging behavior of individuals with intellectual disability. AB - In many Westernized countries, including Australia, concerns about the use of psychotropic drugs to manage the challenging behavior of individuals with intellectual disability have resulted in the development of legislative and procedural controls. Although these constraints may limit indiscriminate use, employing medication remains a common practice. This study examined information about 873 individuals (566 males, 307 females) who were the subjects of reports to the Intellectual Disability Review Panel in March 2000 concerning the use of chemical restraint. A high proportion of people with intellectual disability were reported to have received drugs for purposes of behavioral restraint. The range of drugs was extensive, although those from the antipsychotic class were the most frequently reported. Many individuals concurrently received more than one type of drug or more than one drug from the same drug class. More males than females and more older than younger individuals were administered medication. A relationship between gender and age was apparent, with younger males but older females dominating. The use of drugs to mange the behavior of people with intellectual disability may at times be warranted. However, it is important that the extent and type of drug use, as well as the characteristics of those who are medicated, be subject to ongoing scrutiny. PMID- 15541631 TI - Relationship of preschool special education outcomes to instructional practices and parent-child interaction. AB - Developmental outcomes attained by children receiving preschool special education services in relationship to both the general instructional approach used by their teachers and their parents' style of interaction were examined. The sample included 70 children from 41 Early Childhood Special Education (ECSE) classrooms. The type of instructional model children received was determined by dividing the sample into three clusters based upon six global ratings of children's classroom environment: Choice; Cognitive Problem-Solving; Child-Initiated Learning; Developmental Match; Child-Centered Routines; and Rewards and Discipline Strategies. Based on this analysis, 27 children were classified as receiving developmental instruction; 15 didactic instruction; and 28 naturalistic instruction. Observations of parent-child interaction collected at the beginning and end of the year were classified along four dimensions using the Maternal Behavior Rating Scale: Responsiveness, Affect, Achievement Orientation and Directiveness. Results indicated that the kinds of experiences that children received varied significantly across the three instructional models. However, there were no significant differences in the impact of these instructional models on children's rate of development. Regression analyses indicated that children's rate of development at the end of intervention was significantly related to their parents' style of interaction but was unrelated to the type of instructional model they received. PMID- 15541632 TI - Visual-auditory integration during speech imitation in autism. AB - Children with autistic spectrum disorder (ASD) may have poor audio-visual integration, possibly reflecting dysfunctional 'mirror neuron' systems which have been hypothesised to be at the core of the condition. In the present study, a computer program, utilizing speech synthesizer software and a 'virtual' head (Baldi), delivered speech stimuli for identification in auditory, visual or bimodal conditions. Children with ASD were poorer than controls at recognizing stimuli in the unimodal conditions, but once performance on this measure was controlled for, no group difference was found in the bimodal condition. A group of participants with ASD were also trained to develop their speech-reading ability. Training improved visual accuracy and this also improved the children's ability to utilize visual information in their processing of speech. Overall results were compared to predictions from mathematical models based on integration and non-integration, and were most consistent with the integration model. We conclude that, whilst they are less accurate in recognizing stimuli in the unimodal condition, children with ASD show normal integration of visual and auditory speech stimuli. Given that training in recognition of visual speech was effective, children with ASD may benefit from multi-modal approaches in imitative therapy and language training. PMID- 15541633 TI - Re-evaluation of constant versus varied punishers using empirically derived consequences. AB - Charlop, Burgio, Iwata, and Ivancic [J. Appl. Behav. Anal. 21 (1988) 89] demonstrated that varied punishment procedures produced greater or more consistent reductions of problem behavior than a constant punishment procedure. More recently, Fisher and colleagues [Res. Dev. Disabil. 15 (1994) 133; J. Appl. Behav. Anal. 27 (1994) 447] developed a systematic methodology for predicting the efficacy of various punishment procedures. Their procedure identified reinforcers and punishers (termed "empirically derived consequences" or EDC) that, when used in combination, reduced the destructive behavior of individuals with developmental disabilities who displayed automatically maintained destructive behavior. The current investigation combines these two lines of research by comparing the effects of constant versus varied punishers on the self-injury of two individuals with developmental disabilities. The punishing stimuli were selected via the procedures described by Fisher et al. and were predicted to be at varying levels of effectiveness. The varied presentation of punishers resulted in enhanced suppressive effects over the constant presentation of a punisher for one of two individuals, but only in comparison to a single stimulus predicted to be minimally effective. Even then, the differences were small. These results suggest that the additive effects of varied punishment are negligible if clinicians use stimuli predicted to be effective and are discussed in terms of the conditions under which stimulus variation could potentially enhance the effects of punishers. PMID- 15541634 TI - Epidermal desquamation. AB - Epidermal desquamation, a continuous but insensible bodily activity, is largely ignored unless the rate or amount of scale production becomes abnormal. It is the last topic to be considered in any serious discussion of epidermal growth and differentiation, but is becoming an increasingly fertile ground for investigation. This review summarizes: (a) methods for measuring desquamation; (b) variables that affect normal desquamation; (c) mechanisms of desquamation; (d) the role of desquamation in nutritional homeostasis; and (e) the role of desquamation as a first line of defense. Consideration is given to whether desquamation might be harnessed to eliminate or remediate toxins that have accumulated in the body. PMID- 15541635 TI - The role of pathogenic B-cell clones in antibody mediated autoimmune disorders. AB - Our understanding of the role of B-cells in the immunopathogenesis of certain antibody mediated diseases has developed remarkably in the past few years. In this review, autoantibody mediated immune disorders associated with pathogenic B cell clones are discussed. We have focused on the roles and pathogenic mechanisms of B-cell clones in autoantibodyimmune diseases. The roles of pathogenic B-cells in Castleman's disease in PNP patients is used as one example. The developments in the treatment of B-cell mediated autoimmune diseases, such as intravenous immunoglobulin (IVIg), targets the regulatory pathway of B-cells, using anti CD20, CD19, CD22 and, CD95 monoclonal antibody therapy, etc. are also discussed. Immunotherapy, targeting specific pathogenic B-cells, is believed to be one approach in the management of autoimmune diseases. PMID- 15541636 TI - Two kinds of ENU-induced scant hair mice and mapping of the mutant genes. AB - OBJECTIVE: To establish mouse models for human diseases through N-ethyl-N nitrosourea (ENU) mutagenesis, and to provide groundwork to clone genes and study their functions after mapping the mutant genes. METHODS: 18 male D2 mice (G0) at age of 8-10 weeks old were injected intraperitoneally with ENU (100 mg/kg) once a week for three consecutive weeks. The treated male mice were mated with females of the same strain, and their offspring (G1) were used to screen for dominant and recessive mutation. After breeding the mutant F2 (D2B6 F1 intercrossing) mice, 39 microsatellites that are equally distributed on the mouse genome and are different between B6 and D2 strains were used to scan the genome. According to the log odds score (LODS) we determined whether these microsatellites were linked to the mutant genes and calculated the location of mutant genes based on their recombination ratio. RESULTS: We screened 532 G1 mice, of which 14 exhibited mutation phenotypes. None was dominantly hereditable. Two cases of recessive inheritable scant hair mice were obtained through testing 30 G1 mice with normal phenotype and potential recessive mutant genes. All showed scant coat hair, grew slowly, and hyperkeratoses of epidermis and bollicular horn plug in histological sections. Their visceral organs were not markedly different from normal, and they were named scant hair 1 Baojin (symbol is snthr(-1Bao)) and scant hair 2 Baojin (symbol is snthr(-2Bao)). Through microsatellite screening we found that the LODS between snthr(-1Bao) and D9Mit243 was 7.73, and the linkage was determined. After analyzing the recombination ratio between snthr(-1Bao) and microsatellite D9Mit18 which was near snthr(-1Bao) based on a total number of 126 F2 mice with the scant hair phenotype, we determined that snthr(-1Bao) was located at chromosome 9 and was 71cM from centromere. Using the same technique, snthr(-2Bao) was mapped to the same position as snthr(-1Bao). CONCLUSION: In our research, two cases of scant hair mice provide good models for the study of dermatology, and the location of mutant genes provides a solid foundation for cloning new mice scant hair genes. PMID- 15541637 TI - Gene expression of enzymes for tryptophan degradation pathway is upregulated in the skin lesions of patients with atopic dermatitis or psoriasis. AB - BACKGROUND: Atopic dermatitis (AD) and psoriasis are common inflammatory skin diseases. Although many reports implicate Th2 cytokines in the pathophysiology of AD and Th1 cytokines in psoriasis, the precise etiology of these diseases remains elusive. OBJECTIVE: We investigated novel AD- or psoriasis-related genes to further understand the pathogenesis of these diseases. METHODS: We performed a comprehensive analysis of mRNA expression in skin biopsies from AD or psoriasis patients using DNA microarrays. Quantitative PCR was then used to monitor the expression of novel disease-related genes in human keratinocytes or pinnae from NC/Nga mice. RESULTS: Levels of mRNA for IDO (indoleamine 2,3-dioxygenase) and kynureninase, enzymes constituting the tryptophan degradation pathway, were found to be upregulated in the skin lesions as compared to the uninvolved skin of patients with AD or psoriasis. Expression of these two genes was induced in human epidermal keratinocytes stimulated with IFN-gamma in vitro. Moreover, in NC/Nga mice, the expression of kynureninase mRNA in the ear skin was induced following development of AD-like skin lesions. CONCLUSION: The tryptophan degradation pathway may play an important role in the pathophysiology of AD and psoriasis. PMID- 15541638 TI - Effect of an antiallergic drug (Olopatadine hydrochloride) on TARC/CCL17 and MDC/CCL22 production by PBMCs from patients with atopic dermatitis. AB - BACKGROUND: Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by the predominant infiltration of Th2-type cells in lesional skin. Thymus and activation-regulated chemokine (TARC/CCL17) and monocyte-derived chemokine (MDC/CCL22) are Th2-type cytokines, and it has been reported that serum CCL17 and CCL22 levels are associated with AD disease activity. Olopatadine hydrochloride (Olopatadine) is an antiallergic drug with selective histamine H(1) receptor antagonist activity. The effect of Olopatadine on chemokine production by peripheral blood mononuclear cells (PBMCs) in AD patients has not been completely elucidated. OBJECTIVES: This study was undertaken to clarify the effects of Olopatadine on CCL17 and CCL22 production by PBMCs from patients with AD during the treatment. METHODS: We measured plasma levels of CCL17, CCL22, IFNgamma, IL-12 and IL-18 in 15 patients with AD before and after treatment with oral Olopatadine (10 mg/day) for 4 weeks. We also examined disease activity using SCORAD index, eosinophil numbers in peripheral blood and serum levels of LDH. PBMCs from the patients were taken before and after the treatment and cultured with or without dust mite allergen extract (DME) for 3 or 5 days. CCL17, CCL22, IFNgamma, IL-12 and IL-18 levels in the supernatants of cultured PBMCs were measured. RESULTS: SCORAD index and eosinophil numbers in peripheral blood significantly decreased during treatment of AD patients with oral Olopatadine and topical corticosteroids for 4 weeks. The plasma levels of CCL17 and CCL22 significantly decreased after the treatment compared with before the treatment (p<0.05) and were significantly correlated with SCORAD index. PBMCs from AD patients taken after the treatment and cultured with DME for 5 days, showed significantly lower levels of CCL17 production than those taken before the treatment (p=0.018). PBMCs from AD patients taken after the treatment and cultured with DME for 5 days, also showed significantly lower levels of IFNgamma production than those taken before the treatment (p=0.012). CONCLUSION: Our data demonstrate that Olopatadine inhibits CCL17 and CCL22 production by PBMCs from AD patients, which are important regulators of Th2 recruitment in the skin. PMID- 15541639 TI - Staphylococcus aureus infection of human primary keratinocytes. PMID- 15541640 TI - Expression of androgen receptor, estrogen receptor alpha and beta in the dermal papilla of human hair follicles in vivo. PMID- 15541641 TI - Cutaneous mixed tumors: an immunohistochemical study using two antibodies, G-81 and C8/144B. PMID- 15541642 TI - Dominant Bcl-2 expression during telogen-anagen transition phase in human hair. PMID- 15541643 TI - Effects of MAPK inhibitors on CCR4-mediated chemotaxis against thymus and activation-regulated chemokine (TARC/CCL17). PMID- 15541644 TI - Zinc transporter LIV-1: a link between cellular development and cancer progression. AB - Genes involved in normal developmental processes often have a role in tumour progression. Epithelial-mesenchymal transition (EMT) is a general phenomenon that is an essential event in embryonic development, tissue remodelling and wound repair. Genes that regulate EMT are attracting significant attention as mediators of tumour progression. Indeed, EMT is considered an important event during the evolution of malignant tumours and their progression to metastasis, with several reports of molecules involved in this process facilitating migration and invasion of epithelial tumour cells. Recent work linking the zinc transporter LIV-1 with the transcription factors STAT3 and Snail in zebrafish embryogenesis, throws new light on this transition and has important implications for the development of cancer. PMID- 15541645 TI - The effect of stress on menstrual function. AB - Historically difficult to define, stress is, in one sense, the factor that stressors have in common in their impact on the body. Menstrual function is disrupted by stressors that activate the hypothalamic-pituitary-adrenal (HPA) axis; this activation is part of a catabolic response of the whole body that mobilizes metabolic fuels to meet energy demand. Functional menstrual disorders are associated with an increase in cortisol and with a broad spectrum of other symptoms of energy deficiency. Recent experiments suggest that exercise and other stressors have no disruptive effect on reproductive function beyond the impact of their energy cost on energy availability. These studies suggest that treatments for functional menstrual disorders should aim at dietary reform and that stress is simply low energy availability. Future experiments should carefully test this hypothesis. PMID- 15541646 TI - Puberty, obesity and ethnicity. AB - Lay and medical sources state that children enter puberty earlier now than in past decades. Although it is clear that puberty and menarche currently begin at younger ages than they did in past centuries, a comparison of the onset of puberty during the few past decades is more difficult to interpret. African American girls do start puberty earlier and menarche is earlier than in White girls, and the gap between the two has widened during the past 20 years. This change between ethnic groups might be due to an increasing difference in body mass index (BMI) values. Greater BMI values are associated with pubertal onset or menarche; thus, if BMI continues to increase in childhood, earlier puberty might well be found in the general population in the future. In the absence of accurate data from the past or definitive studies at present, however, it cannot be stated with assurance that puberty today starts significantly earlier than it did in the past 4-5 decades across the whole population. PMID- 15541647 TI - Animal models of preterm birth. AB - Preterm birth continues to pose a significant clinical dilemma and contributes to both acute and long-term neonatal morbidity. Despite efforts, the incidence of preterm birth has not decreased, partly because of our lack of understanding of the mechanisms that trigger parturition. Animal models are essential research tools for investigating the pathways that promote preterm parturition and for testing therapeutic interventions. Growing evidence correlates infection or inflammation with preterm birth. Consequently, many investigators have created animal models that reflect these findings. Current models of preterm parturition include diverse species, varying means of inducing an inflammatory or infectious state, and different routes of administration. Although each of these models can advance our knowledge, it is important to understand their advantages, disadvantages and unique characteristics. An understanding of such models will hopefully promote continued research that will ultimately lead to a decrease in preterm birth and an improvement in neonatal outcome. PMID- 15541648 TI - The electrophysiology of feeding circuits. AB - Obesity is quickly becoming one of the most common and debilitating disorders of the developed world. More than 60% of American adults are now overweight or obese, predisposing them to a host of chronic diseases. To understand the etiology of obesity, and to discover new therapies for obesity, we must understand the components of energy balance. In simple terms, energy intake (feeding) must equal energy expenditure (physical activity, basal metabolism and adaptive thermogenesis) for body weight homeostasis. To maintain homeostasis, neurocircuitry must sense both immediate nutritional status and the amount of energy stored in adipose tissue, and must be able to provide appropriate output to balance energy intake and energy expenditure. The brain receives various signals that carry information about nutritional and metabolic status including neuropeptide PYY(3-36), ghrelin, cholecystokinin, leptin, glucose and insulin. Circulating satiety signals access the brain either by "leakage" across circumventricular organs or transport across the blood-brain barrier. Signals can also activate sensory vagal terminals that innervate the whole gastrointestinal tract. PMID- 15541649 TI - Role of PPARgamma in macrophage biology and atherosclerosis. AB - Macrophages carry out key functions by defending a host from microbial invaders and by clearing endogenous cellular debris. Molecules that are essential for the recognition, phagocytosis and clearance of pathogens also mediate the uptake and degradation of pathogenic lipoproteins. During atherogenesis, for example, scavenging trapped lipoproteins leads to the formation of foam cells and subsequently the activation of these lipid-laden macrophages. Although they are initially clinically silent, these fatty streaks evolve into complex inflammatory plaques that cause significant morbidity and mortality. Thus, interventions that decrease foam cell formation and reduce the inflammatory response of macrophages could become effective therapies for coronary artery disease. Thiazolidinediones (TZDs) might be developed as anti-atherogenic agents on the basis of their actions as ligands for peroxisome proliferator-activated receptor-gamma (PPARgamma). PMID- 15541650 TI - Adaptors in immune regulation. PMID- 15541651 TI - Regulation of the immune system by SOCS family adaptor proteins. AB - Signal transduction via cytokine receptors is regulated by several mechanisms that control initiation, magnitude and duration of the signaling pathways. Cytokine-induced SOCS family adaptors function as feedback inhibitors of cytokine receptor signaling by inhibiting the JAK-STAT signal transduction pathway. Specific gene-targeted mice have unveiled critical, non-overlapping functions for SOCS1 and SOCS3 in lymphocyte development and homeostasis, and in the regulation of macrophage and dendritic cell functions. In this review, we will discuss the structure of SOCS proteins, mechanisms by which they control the JAK-STAT pathway and their role in immune regulation. PMID- 15541652 TI - Transmembrane adapters: structure, biochemistry and biology. AB - Transmembrane adapter proteins (TRAPs) represent a relatively new and unique group of signalling molecules in hematopoetic cells. They differ from other signalling proteins as they lack any enzymatic or transcriptional activity, instead they possesses multiple tyrosine-based signalling motifs (TBSMs). Triggering of immunoreceptors induces tyrosine phosphorylation of these motifs by members of the Src-, Syk- or Tec-family of protein tyrosine kinases thus enabling the TRAPs to recruit cytosolic adapter and/or effector molecules via their SH2 domains into close proximity to the immunoreceptors, a position from which they can coordinate and modulate signal transduction pathways important for lymphocyte function. PMID- 15541653 TI - The SLP-76 family of adapter proteins. AB - Adapter molecules are multidomain proteins lacking intrinsic catalytic activity, functioning instead by nucleating molecular complexes during signal transduction. The SLP-76 family of adapters includes SH2 domain-containing leukocyte phosphoprotein of 76kDa (SLP-76), B cell linker protein (BLNK), and cytokine dependent hematopoietic cell linker (Clnk). These proteins are critical for integration of numerous signaling cascades downstream of immunotyrosine-based activation motif (ITAM)-bearing receptors and integrins in diverse hematopoietic cell types. Mutations in genes encoding SLP-76 family adapters result in severe phenotypes, underscoring the critical role these proteins play in cellular development and function by directing formation of signaling complexes in a temporally- and spatially-specific manner. PMID- 15541654 TI - Involvement of the Wiskott-Aldrich syndrome protein and other actin regulatory adaptors in T cell activation. AB - The actin cytoskeleton is a dynamic structure recognized for many years as integral to the coupling of external stimuli to cell activation and ensuing changes in morphology and movement. It is only recently, however, that a molecular understanding of actin involvement in these activities has emerged coincident with the identification of cytosolic signaling effectors that couple extracellular stimuli to induction of actin nucleation. Notable among these actin regulatory effectors are members of the Wiskott-Aldrich syndrome protein (WASp) family, a group of cytoskeletal adaptors imbued with the capacity to connect various signal transduction pathways to the Arp 2/3 complex and Arp 2/3-mediated actin polymerization. In T cells, the functional characterization of WASp and other actin-modulatory adaptors has proved instrumental in delineating the molecular interactions evoking actin cytoskeletal reorganization downstream of antigen receptor engagement and in clarifying the influence of actin-based processes on T cell activation. In this review, the structural and functional properties of the major actin regulatory cytoskeletal adaptors in T cells are described with an emphasis on the roles of these proteins in fostering the TCR actin cytoskeletal interplay required for induction of T cell activation and expression of dynamic effector responses. PMID- 15541655 TI - The SAP family of adaptors in immune regulation. AB - SAP and EAT-2 define a new class of adaptor proteins composed almost exclusively of a Src homology 2 (SH2) domain. By way of their SH2 domain, SAP-like adaptors interact with tyrosine-based motifs in the cytoplasmic region of SLAM-related receptors, a family of immune cell-specific molecules involved in immunoregulation. Recent findings indicate that SAP is required for the functions of SLAM family receptors, as a consequence of its ability to promote recruitment of Src-related protein tyrosine kinase FynT and allow SLAM-related receptors to transduce tyrosine phosphorylation signals. SAP is mutated in X-linked lymphoproliferative (XLP) syndrome, a rare inherited human disease characterized by an deregulated immune response to Epstein-Barr virus infection. Several lines of evidence indicate that defects in the activities of SLAM-related receptors caused by SAP deficiency account for the immune dysfunctions associated with XLP. PMID- 15541656 TI - Inhibitory adaptors in lymphocytes. AB - The negative regulation of lymphocyte activation and function is mediated by inhibition of signaling through antigen-receptor, co-stimulation receptor or cytokine receptor. The suppression of downstream signaling through antigen receptor is mediated by negative regulators including adaptors and effectors such as phosphatases. "Inhibitory adaptors" exhibit their inhibitory function directly or indirectly by the localization to the vicinity of the antigen-receptor on the membrane. The strategy of inhibition by inhibitory adaptors includes the recruitment of inhibitory effector molecules, sequestration of positive regulators, internalization/degradation of receptor complexes, and the blockade of the dynamic movement of positive regulators. PMID- 15541657 TI - The roles of CARMA1, Bcl10, and MALT1 in antigen receptor signaling. AB - Lymphocyte activation plays a critical role in immune responses. Dysregulation of lymphocyte activation can cause autoimmune, immunodeficient diseases, or leukemia/lymphoma. Lymphocyte activation is triggered by stimulation of antigen receptors, T cell receptors (TCR) or B cell receptors (BCR), on the surfaces of T or B lymphocyte, respectively. Stimulation of TCR or BCR induces a series of signal transduction cascades leading to activation of multiple transcription factors including NF-kappaB. Recent studies demonstrate that CARMA1, a scaffold protein, plays an essential role in mediating TCR- or BCR-induced NF-kappaB activation by recruiting two adaptor proteins, Bcl10 and MALT1, to lipid rafts following stimulation of antigen receptors. In this review, we will discuss the mechanism by which proximal signaling components connect antigen receptor signaling to CARMA1, and how CARMA1 regulates Bcl10 and MALT1, leading to activation of NF-kappaB. In addition, the roles of CARMA1, Bcl10, and MALT1 in lymphocyte activation and development will also be discussed. PMID- 15541659 TI - History and evolution of disc replacement. AB - Total joint replacement has revolutionized the treatment of limb arthritides. Spinal arthroplasty is emerging as a treatment for spinal disc degeneration. The purpose of this review is to highlight the evolution of artificial interventions for nucleus and total disc replacement. This review will provide the foundation for the understanding of present and future technologies in the field. PMID- 15541660 TI - Why a mechanical disc? AB - Low back pain secondary to degenerative disc disease is an overwhelming and growing problem in the United States and Western countries. Most degenerative disc disease can be treated nonoperatively. There are, however, substantial numbers of patients who have not benefited from exhaustive nonoperative treatments and subsequently seek surgical solutions to their incapacitating back pain. Lumbar fusion for back pain and/or leg pain associated with degenerative disc disease is considered the gold standard by which other treatments are judged. A challenge to spinal fusion for degenerative disc disease is now being offered in the form of the artificial disc. The implantation of an artificial lumbar disc allows for maintenance or restoration of physiologic movement at affected segments. A major long-term complication of spinal fusion is degeneration of a disc adjacent to the fused segments. Theoretically, the maintenance of motion could minimize development of adjacent disc degeneration as seen with spinal fusion. It is interesting to note that fusion of the hip or knee is not considered a primary procedure, but fusion is a primary procedure for the lumbar spine. Four artificial lumbar discs are discussed in this article. Early results are promising in terms of clinical results and movement, but long-term follow-up clinical trials must be done in order to gain an accurate comparison with spinal fusion. Trials are currently ongoing. The clinical results up to now and the potential for maintaining lumbar mobility throughout life warrant continuation of this surgical procedure. PMID- 15541661 TI - Molecular basis of intervertebral disc degeneration. AB - This review will acquaint the reader with normal development, structure, and function of the intervertebral disc. The disc is composed of the vertebral endplate, nucleus pulposus, and annulus fibrosus. These three functional components all serve important purposes for health and function of the disc. Nutrition and biomechanics are also discussed. The molecular basis of disc degeneration is reviewed so that biologic approaches to the reversal and or treatment of disc degeneration may be better understood. PMID- 15541662 TI - A new classification system for degenerative disc disease of the lumbar spine based on magnetic resonance imaging, provocative discography, plain radiographs and anatomic considerations. AB - Prior attempts to classify degenerative disc disease (DDD) of the lumbar spine have been based on magnetic resonance imaging (MRI) signal intensity. Internal disruption of the disc is not reliably diagnosed by MRI alone. No attempt has been made to include provocative discography and plain radiographs. The intervertebral joint is a three-joint complex consisting not only of the end plate-disc-end plate joint of the anterior column, but also the two facet joints of the posterior column. To date, no classification system for lumbar DDD has taken into account the state of degeneration of the facet joints. In the past this may not have been important with regard to arthrodesis. But as early experience indicates, the degree of degeneration of the facet joints is paramount to the clinical success of an artificial disc replacement. As we transition from intervertebral joint fusion to intervertebral joint replacement, it is imperative that the state of degeneration of the three-joint complex be classified in such a way that all can evaluate lumbar DDD using the same terminology. A new classification system for lumbar DDD is presented. The intervertebral segment is graded in two parts: the anterior column and the posterior column, based on MRI, discography and plain radiographs. The system will be validated through a blinded study of 100 patients requiring stand-alone anterior lumbar interbody fusion or circumferential fusion. Although the debate over which discs are best treated surgically with indication for total disc replacement, or with indication for fusion will continue for some time, the basic understanding of intervertebral disc and posterior facet joint pathology, classified and validated in a simple forthright manner, is essential as we move forward with artificial disc technology. PMID- 15541663 TI - A review of spinal fusion for degenerative disc disease: need for alternative treatment approach of disc arthroplasty? AB - This article reviews the history of spinal fusion. The most common indication for fusion may be painful disc degeneration although the exact determination of this condition remains controversial. In the analysis of surgical fusion techniques, literature review has documented a higher rate of success in attaining successful fusion than in obtaining excellent clinical outcomes. Significant morbidity can be seen in the performance of spinal fusion. A limited number of patients presently undergoing spinal fusion will be candidates for disc arthroplasty, though indications for spinal arthroplasty are still evolving. PMID- 15541664 TI - The indications for lumbar and cervical disc replacement. AB - Although cervical and lumbar disc replacements are in their spinal surgical infancy with regard to clinical application, the indications are already delineated. Lumbar disc arthroplasty is indicated for one- or two-level discogenic mechanical back pain primarily in the absence of radiculopathy. In contrast, cervical disc replacement can be readily applied in patients presenting with neurologic deficit, radiculopathy or myelopathy because the approach and anterior spinal cord decompression are identical for anterior cervical disc replacement and traditional Smith-Robinson cervical decompression. In addition, the application of more complex spinal osteotomies, revision of pseudarthroses and deformity correction are much more applicable to the cervical arthroplasty procedures. This is because even the most experienced vascular access surgeon has difficulty with the formidable revision through a repeat anterior lumbar procedure, whereas most experienced cervical spinal surgeons are familiar with repeat anterior cervical approaches. The end result is that surgeons will have more trepidation with multilevel lumbar arthroplasties, especially those presenting in conjunction with neurologic symptoms. At the current time radiculopathy is an exclusion criteria for the four prospective Investigational Device Exemption (IDE) Food and Drug Administration (FDA) studies on lumbar disc replacement, whereas cervical radiculopathy is an inclusion criteria for the major IDE FDA investigations of cervical arthroplasties. PMID- 15541665 TI - Disc arthroplasty: lessons learned from total joint arthroplasty. AB - BACKGROUND CONTEXT: With the rapid emergence of new disc arthroplasty designs and ongoing clinical trials, there is great anticipation among clinicians. Inevitably there will be challenges associated with adoption of this new technology. Potential lessons learned from the extensive experience with extremity arthroplasty may predict some of the perils and pitfalls associated with motion preserving devices in spinal applications. PURPOSE: To review the extremity total joint literature for lessons that may be transferable to disc arthroplasty and anticipate predictable problems that will occur. STUDY DESIGN/SETTING: A literature review and conjectural analysis of key lessons learned from the experience of extremity total joint arthroplasty and anterior lumbar interbody fusion devices with subsequent application of these lessons to disc arthroplasty in the lumbar spine. METHODS: Limited data are available on disc arthroplasty. RESULTS: Particular areas of controversy and uncertainty exist. The ongoing US Food and Drug Administration Investigational Device Exemption clinical trials may provide additional insight into these questions. CONCLUSIONS: Complications will occur. These will be approach related, device related and patient-selection related. Studying the experience of extremity joint arthroplasty may accelerate the learning curve for lumbar disc arthroplasty. PMID- 15541666 TI - Adjacent segment degeneration and adjacent segment disease: the consequences of spinal fusion? AB - Given the number of spinal fusions performed annually, concerns have mounted over the potential for adjacent segment degeneration (radiographic changes of degeneration at levels adjacent to a spinal fusion) and adjacent segment disease (development of new symptoms correlating with adjacent segment degeneration). This article reviews documented evidence on adjacent segment degeneration and disease as it relates to cervical and lumbar arthrodesis. There appears to be an incidence of adjacent segment degeneration and disease after arthrodesis that may be related to natural degeneration or the adjacent fusion. It remains to be seen whether restoration of motion with disc arthroplasty will alter the rate of adjacent segment degeneration or disease. PMID- 15541667 TI - Assessing the potential impact of total disc arthroplasty on surgeon practice patterns in North America. AB - BACKGROUND CONTEXT: Intervertebral disc replacement has enjoyed a recent surge in popularity largely because of its anticipated release into the North American market. These devices offer an alternative to the 450,000 lumbar and cervical fusion procedures performed by US spine surgeons each year. The unanswered question at the present time is the potential impact, technologically and economically, of cervical and lumbar disc arthroplasty on the practice patterns of North American spine surgeons. PURPOSE: To assess the economic and societal impact of motion-sparing technology in the North American Market. SETTING: A review of the literature and an economic market analysis of the impact of intervertebral disc replacements on surgeon practice in North America. CONCLUSION: The potential economic effect of spine arthroplasty in the United States is staggering with conservative figures approximating $2.18 billion and 47.9% of the market share being captured by motion-sparing technology. Modeling spinal arthroplasty usage and influence after other spinal products, such as cylindrical cages and pedicle screws, may provide insight into the effects of motion-sparing technology. Regardless of the clinical outcome of disc arthroplasty, its potential economic impact will be significant. PMID- 15541668 TI - Degenerative mechanics of the lumbar spine. AB - This paper reviews both the normal biomechanical behavior of the lumbar motion segment and changes in that behavior with disc degeneration. Each level of the spine consists of a disc anteriorly and two facet joints posteriorly. A complex ligamentous structure also contributes to the mechanics of the spine. Degenerative changes include intervertebral disc (nuclear) degeneration, facet joint osteoarthritis, vertebral body degeneration, and ligament degeneration. The goal of this review is to discuss the effect of disc degeneration on the biomechanical behavior of the spine, specifically the load-bearing and kinematic behavior after degeneration. PMID- 15541669 TI - Artificial disc prosthesis: design concepts and criteria. AB - Clinical and biomechanical objectives of disc arthroplasty are reviewed in this paper. The available literature relative to these criteria for nucleus and total disc replacement where relevant is also reviewed. Clinical criteria include: pain relief, functional recovery, lower morbidity than spinal fusion, shorter recuperation, and ease of implantation and revision. Biomechanical criteria include: preservation of motion in compression-bending and compression-torsion, shock absorption, relief of abnormal stresses within and adjacent to the motion segment, stability, and alignment. A review of devices with these criteria in mind is reported. PMID- 15541670 TI - Basic scientific considerations in total disc arthroplasty. AB - BACKGROUND CONTEXT: Total disc arthroplasty serves as the next frontier in the surgical management of intervertebral discogenic pathology. PURPOSE: As we move from an era of interbody spinal arthrodesis to one in which segmental motion is preserved, this promising new technology offers increasing clinical and research challenges in the areas of spinal kinematics, histologic osseointegration at the prosthetic-bone interface and the effects of particulate wear debris. STUDY DESIGN: The primary focus of this paper is to provide a methodologic basis to investigate the spinal kinematics, histologic osseointegration and particulate wear debris after total disc arthroplasty by using in vitro and in vivo models. METHODS: Part I: Using an in vitro cadaveric model, multidirectional flexibility testing evaluated the functional unit kinematics under the following L4-L5 reconstruction conditions: 1) intact spine, 2) Charite disc prosthesis, 3) BAK cages, 4) BAK cages+ISOLA pedicle screw or rod fixation (anteroposterior). Part II: A total of 27 mature baboons (n=27, Papio cynocephalus) underwent L5-L6 total disk replacement procedures to investigate the biomechanical, histochemical and biologic ingrowth characteristics of two different lumbar disc prostheses (AcroFlex and Charite) for total disc arthroplasty. Functional spinal unit fusion status was assessed by using radiographic analysis, biomechanical testing, undecalcified histopathologic and histomorphometric analyses. Part III: Using a total of 50 New Zealand white rabbits, this investigation served to quantify the neural and systemic tissue histopathologic response, after epidural application of four different types of spinal instrumentation particulate wear debris: 1) sham (control) (n=10), 2) stainless steel 316LVM (n=10), 3) titanium alloy Ti-6AL 4V (n=10), 4) cobalt chrome alloy (n=10) and 5) ultrahigh molecular weight polyethylene (UHMWPE) (n=10). RESULTS: In vitro multidirectional flexibility testing demonstrates the operative and adjacent level motion-preserving properties of total disc arthroplasty versus interbody arthrodesis cages and pedicle screw spinal instrumentation. To this end, disc replacement preserves the normal centrode or locus of intervertebral rotation at the operative and adjacent intervertebral spinal levels compared with conventional stabilization implants. On the basis of nonhuman primate modeling in the current studies, porous titanium interface surfaces afforded the greatest percentage of trabecular ingrowth at the prosthesis-end plate interface. In vivo segmental motion under multidirectional testing was preserved with the Charite device and slightly diminished with the AcroFlex implants. The porous ingrowth coverage at the bone-metal interface was more favorable for total disk replacement (range, 40% to 50%) compared with that reported for cementless total joint components in the appendicular skeleton (range, 10% to 30%). Direct epidural application of spinal instrumentation particulate wear debris elicits a chronic histiocytic reaction localized primarily within the epidural fibrous layers. Moreover, particles have the capacity to diffuse intrathecally, eliciting a macrophage and cytokine response within the epidural tissues, cerebrospinal fluid and spinal cord itself. Overall, on the basis of the postoperative time periods evaluated, no evidence was observed of an acute neural or systemic histopathologic response to the materials included in the current project. CONCLUSION: The implementation of dynamic spinal stabilization systems for fusionless correction of spinal deformity, dynamic posterior stabilization and total disc arthroplasty necessitates improved understanding with regard to spinal kinematics, patterns and mechanisms of histologic osseointegration and the neurohistopathologic response to particulate wear debris. Collectively, the current studies provide a methodologic basis to comprehensively evaluate these three areas. PMID- 15541671 TI - Material considerations for intervertebral disc replacement implants. AB - Cervical and lumbar disc replacements are being performed with increasing frequency. Much of the background for the development for these implants is drawn from the literature of other joint replacements that have been in evolution and use for decades. Important variables for the function and longevity of such disc arthroplasty implants are clearly defined by the material properties of the components used for their production. The most frequently considered materials are cobalt-chrome alloys, titanium alloys, stainless steels, polyethylene, polyurethane and ceramics. In addition to implant materials, the interfaces of such materials must be considered. The bearing surfaces of an implant, in particular, are at risk of wear and failure. Overall, successful, long-term total disc arthroplasty requires a thorough understanding of biomaterials and how they can be used to achieve their desired goals. PMID- 15541672 TI - Evaluation of the biological response of wear debris. AB - STUDY DESIGN: An animal study was conducted to evaluate the biological response to titanium particles from an artificial intervertebral disc in terms of serology and histologic changes. OBJECTIVES: To determine the biological response to wear debris in the retroperitoneal and epidural space. SUMMARY OF BACKGROUND DATA: Few wear studies exist about mechanical artificial discs. METHODS: Twenty-three New Zealand white rabbits were used for two approaches of the lumbar spine. In a retroperitoneal group (10 rabbits), lateral flank approach at the L2-L3 area was used. In an epidural group (13 rabbits), a dorsal laminotomy of L2 was performed. The wear debris was obtained from mechanical test cycling of the implantable intervertebral disc. At 4 and 12 weeks postoperatively, five or six animals from each group were killed. The tissues, including deposition site, regional lymph nodes and major organs, were evaluated with hematoxylin and eosin staining. RESULTS: At death all rabbits were found to be healthy. Blood results from the predeath samples were found to be consistent with the preoperative blood work values. Scar tissue was minimal with good healing. All organs were found to be normal in appearance. On histopathology sections, adverse reactions such as fibrosis, granuloma formation or necrosis were not found in any tissues. Free particles were found sparingly in all tissue sections with minimal cellular response. No remarkable difference was found according to groups or time intervals. Smaller particles were found to be engulfed in macrophages without adverse biological consequences. CONCLUSION: Titanium particles traveled from the sites of deposition but elicited no to minimal biological response. PMID- 15541673 TI - AcroFlex design and results. AB - The AcroFlex prosthesis is unique when compared to other current devices, as it has an elastomeric core sandwiched between two titanium endplates. The purpose of this design was to optimize shock absorption qualities. This paper reviews the design of the AcroFlex Disc and relevant design and outcomes of two pilot studies carried out on 28 patients using the AcroFlex prosthesis. Although clinical outcomes of the pilot studies were successful, the pivotal randomized trial has not been carried out due to detection of mechanical failure of the elastomer on thin cut CT scans. The implications of this for the AcroFlex and other arthroplasties are discussed. PMID- 15541674 TI - Prospective randomized study of the Charite artificial disc: data from two investigational centers. AB - BACKGROUND CONTEXT: For decades there has been a desire to restore motion of a painful degenerated spinal segment. Artificial discs have been used in Europe for almost 20 years. In the few reports available in the literature, the results have been promising. However, there have been no prospective randomized studies comparing artificial discs with spinal fusion. PURPOSE: The purpose of this study was to determine if patients with symptomatic degenerative disc disease treated with Charite artificial disc (DePuy Spine, Raynham, MA) arthroplasty would show significant improvement in functional outcome measures and to compare these results to fusion. STUDY DESIGN/SETTING: This was a prospective randomized clinical trial comparing total disc replacement with anterior lumbar interbody fusion using cages. The data reported were collected from two spine specialty centers participating in a Food and Drug Administration regulated trial. METHODS: A consecutive series of 144 patients were randomized using a 2:1 ratio of Charite versus BAK (Zimmer Spine, Minneapolis, MN). All patients were being treated for single-level discogenic pain confirmed by plain radiography, magnetic resonance imaging and provocative discography. Data were collected at designated follow-up periods for up to 24 months. RESULTS: The mean age was 40.1 years (range, 21 to 56 years). Forty-four cases had BAK anterior interbody fusion, and 100 cases were randomized to Charite disc replacement. The mean operating time was 76.2 minutes (range, 54 to 137 minutes) for the Charite cases. The mean estimated blood loss was 196.2 cc (range, 50 to 1,800 cc). Most patients were discharged in 1 to 2 days with a soft corset and returned to normal activities within 3 weeks if they underwent the disc replacement. The mean Oswestry Disability Index score for the BAK group was 69.6+/-12.8 preoperatively and 27.5+/-26.4 at 24-month follow-up (p<.001) The corresponding mean Oswestry score for the Charite disc patients was 70.9 preoperatively and 30.0 at 24-month follow-up (p<.001). CONCLUSIONS: In this prospective randomized study, both surgical groups improved significantly. Complications of total disc replacement were similar to those encountered with anterior lumbar interbody fusion. Total disc replacement appears to be a viable alternative to fusion for the treatment of single-level symptomatic disc degeneration unresponsive to nonoperative management. PMID- 15541675 TI - Lumbar spine arthroplasty using the ProDisc II. AB - The ProDisc was developed by Thierry Marnay, a French orthopedic surgeon, in the late 1980s. Marnay and his associate implanted ProDiscs into 64 patients from 1990 to 1993. Demonstrating remarkable intellectual restraint, he let this cohort of patients "incubate" so he could see their progress before performing any additional implantations. In 1998, 61 of these patients were still alive, and 58 (95%) of them were available for a thorough follow-up evaluation. No device related safety issues were identified during this review, and 93% of these patients were satisfied with their implants. This unique and strong data set led to an unusually rapid Food and Drug Administration (FDA) approval for an Investigational Device Exemption study in the United States, prospectively comparing implantation of a ProDisc with a 360-degree fusion, with both single- and double-level study arms. This article represents data on the first 78 patients with at least 6-month follow-up, with 54 of these patients also having 1 year follow-up, enrolled in a prospective randomized FDA study evaluating the safety and efficacy of ProDisc II versus control, a 360-degree lumbar spinal fusion. Data were collected preoperatively at 6 weeks, 3 months, 6 months and 1 year postoperatively. Visual analog scale (VAS), Oswestry Low Back Pain Disability Questionnaire (ODQ) and patient satisfaction rates were evaluated at these intervals, as well as range of motion, return to work, recreational status and ambulatory status. At 6-month follow-up, there were there were 55 ProDisc patients and 23 who underwent fusion. Twenty-five of these patients had two-level surgery. Estimated blood loss (ProDisc, 103 cc, vs fusion, 213 cc) and operative time (ProDisc, 90 minutes, vs fusion, 232 minutes) were significantly different (p< .01). Hospital stays were shorter (ProDisc, 2.24 days, vs fusion, 3.26 days [p< .01]) for ProDisc patients. There was a significant reduction in the ODQ scores from preoperative values in both ProDisc and fusion groups. Similarly, there was a significant reduction for both groups in VAS scores from before to after surgery. A trend was identified at 6 months in patient satisfaction rates favoring ProDisc versus fusion (p= .08), which became more pronounced (although still not statistically significant) at 1 year. Flexion and lateral bend range of motion was significantly improved in ProDisc patients compared with the fusion group (p= .02). Ambulatory status as well as recreational activity improved faster in the ProDisc group. The data suggest that total disc arthroplasty may be an attractive option to lumbar fusion for the surgical treatment of disabling mechanical low back pain secondary to lumbar disc disease. PMID- 15541676 TI - Design rationale and biomechanics of Maverick Total Disc arthroplasty with early clinical results. AB - This paper reviews the design criteria, biomechanical and biological (wear and safety) testing of this chrome cobalt metal-on-metal, ball and socket design prosthesis. The surgical technique and early clinical results of the initial implantations are also reviewed. Initial results of 7 Maverick implantations showed all 7 patients attaining a 15 point Oswestry improvement within 3 months after implantation. This early result in a small sample is significantly quicker in recovery and improvement when compared to the historical control of the LT cage with Infuse IDE study. Longer term results and more careful study are needed of this interesting and optimistic finding. PMID- 15541677 TI - Design and evaluation of the FlexiCore metal-on-metal intervertebral disc prosthesis. AB - BACKGROUND CONTEXT: The technical difficulties associated with the development of an intervertebral disc prosthesis include endurance demands on the device, lack of consensus concerning the biomechanical principles governing the articulation of the spinal joint and the performance of materials available for implantation. PURPOSE: Although biologically based disc prostheses and augmentations may be the endpoint of spinal disc replacements, these devices and associated technologies will still require decades of work in order to achieve fruition. The more immediate solution will require a durable, biocompatible device capable of restoring range of motion. The evaluation of such a device must include failure testing of critical components as well as a series of fatigue experiments under overloaded conditions. STUDY DESIGN/SETTING: Recent literature citing adjacent level degeneration associated with segmental mobilization and a lack of correlation between successful fusion and clinical success has prompted the need for a dynamic intervertebral disc prosthesis. METHODS: Combined with a better understanding of the biomechanics and the prospect of an increasing percentage of more elderly patients, the future of spinal fusion for pain and instability may need to be reexamined. The authors propose a novel metal-on-metal design for an intervertebral device that features a fixed center of rotation, a mechanical torsional limit, a unique feature to allow for the location of the device in a patient-specific manner, and means by which the device may be implanted directly anterior or anterolaterally. RESULTS: Physiological ranges of motion are retained by the prosthesis. In addition, initial and long-term fixations are achieved through spines and bone ingrowth coating. The device comprises a retained ball and socket positioned between two baseplates. The ball and socket joint permits articulation through the appropriate physiological range of motion and center of rotation as the baseplates provide a stable platform for implantation. CONCLUSIONS: An intervertebral disc prosthesis has been designed and has demonstrated mechanical performance beyond what is required physiologically under preliminary testing. PMID- 15541678 TI - Complications and surgical revision for failed disc arthroplasty. AB - This review will outline the indications and options for revision of failed disc arthroplasty. Such indications may include implant loosening, malposition, displacement, early wear, and infection. Each indication will likely require different preoperative planning, testing, and strategies. PMID- 15541679 TI - Goals of cervical disc replacement. AB - The goals of cervical arthroplasty are reviewed against a backdrop of adjacent segment disease, restoring normal kinematics to the motion segment, and avoidance of fusion in situations where fusion may be difficult to obtain or unwanted. PMID- 15541680 TI - Choosing a cervical disc replacement. AB - BACKGROUND CONTENT: Three important basic scientific studies are presented that measured the volumetric density of longitudinal bony columns within the cervical vertebra. The most solid bone is lateral, adjacent to the uncovertebral joints in a radial pattern. PURPOSE: To characterize the best footprint, profile and biomaterials to construct a cervical disc replacement. STUDY DESIGN: A compilation of biomechanical and anatomical basic scientific studies. METHODS: Microcomputed tomographic imaging, trabecular density and mineral distribution were quantitated from human cervical vertebra. RESULTS: The lateral portions of the cervical vertebra are subjected to higher bending loads than the lumbar vertebral bodies. Therefore, the optimal prosthesis needs to be anchored in the lateral uncovertebral bone. To reduce the incidence of cervical subsidence, the prosthesis needs to be more rectangular than round to take advantage of the radially oriented lateral trabeculae. TiCaP (titanium/calcium phosphate) (Cervitech, Inc., Rockaway, NJ) bony ingrowth coating leads to 10% to 15% greater bony integration than plasma-sprayed titanium. TiCaP causes a supersaturated solution of CaP at the metal-bone interface, which enables reprecipitation of hydroxyapatite and superior bony integration. The optimum pore size of the ingrowth coating of the lumbar spine is 75 to 300 microns, whereas in the cervical spine the optimal ingrowth coating is 20 to 30 microns. This is an order of magnitude lower in pore size to match the smaller cervical trabecular architecture. CONCLUSIONS: Kinematic considerations for the cervical spine show the load is 1/9th the load carried by the lumbar spine or 50 N per segment. Knowing the sliding distance and wear characteristics of conventional biomaterials (ultrahigh molecular weight polyethylene and cobalt chrome) demonstrates that the generation of particulate debris should be a very minor consideration with cervical arthroplasty. PMID- 15541681 TI - The Bryan Cervical Disc: wear properties and early clinical results. AB - BACKGROUND CONTEXT: The rationale for motion preservation by disc replacement after anterior cervical discectomy is to diminish long-term morbidity secondary to adjacent segment degeneration. However, these disc prostheses will be subject to wear and its possible adverse consequences. PURPOSE: Assess the in vitro and in vivo wear properties of the Bryan Cervical Disc and the early clinical results. STUDY DESIGN: In vitro mechanical testing, caprine animal model and prospective European human trial. OUTCOME MEASURE: In vitro mechanical testing used American Society for Testing and Materials standards to measure wear rates and debris. Caprine results were analyzed by histopathologic analysis by a blinded veterinary pathologist. Clinical outcomes were measured by Odoms's criteria, independent radiologic analysis and Short Form-36. METHODS: In vitro wear testing in cervical spine simulator for up to 40 M cycles. Wear rates were determined and particles analyzed microscopically. The in vivo inflammatory response was studied in goats that had discectomy at C4-C5. They were sacrificed between 3 and 12 months and histopathologically were compared with controls who had fusions with titanium plates. A prospective clinical trial of 136 patients with minimum 1-year follow-up was analyzed using validated questionnaires and radiographs. RESULTS: In vitro wear averaged approximately 1.76% by weight at 10 M cycles and 18% at 40 M cycles. Wear debris were present in the periprosthetic tissues in 4 of 11 animals without inflammatory response. The early clinical results were satisfactory in over 90% of patients. CONCLUSION: The in vivo and in vitro wear properties are satisfactory for the expected duration of life of the prosthesis. The early clinical results are satisfactory and equal to fusion. PMID- 15541682 TI - The Prestige cervical disc replacement. AB - BACKGROUND CONTEXT: Anterior cervical discectomy and arthrodesis is the preferred treatment for a large number of patients with symptomatic cervical spondylosis. This management strategy is successful for the majority of patients, and the short-term results are very good. There is the potential to further improve patient outcome and the long-term success rate. PURPOSE: The purpose of this report is to review the rationale for cervical arthroplasty and describe the development of the Prestige cervical disc replacement (Medtronic Sofamor Danek, Inc., Memphis, TN). STUDY DESIGN/SETTING: This report reviews the literature concerning the need for cervical arthroplasty and details the basic and clinical work on the Prestige cervical disc replacement. METHODS: The key publications concerning the shortcomings of cervical arthrodesis and the clinical and basic studies on the Prestige cervical disc implant are discussed. New long-term clinical data on the Prestige device are reported. RESULTS: Cervical arthrodesis perturbs normal cervical spinal biomechanics and appears to be associated with accelerated degeneration of the adjacent segments. The Prestige cervical disc replacement is successful in preserving normal spinal motion, both experimentally and clinically. The device is durable, and the clinical outcomes have been excellent. CONCLUSIONS: The Prestige cervical disc replacement is an effective means of reconstructing the cervical spine after anterior discectomy. It preserves normal motion and may decrease the incidence of adjacent segment disease. PMID- 15541683 TI - Clinical experience with the new artificial cervical PCM (Cervitech) disc. AB - The results of a pilot study performed between December 2002 and October 2003 in which 82 cervical disc arthroplasties were implanted in 53 patients are reviewed in detail. Visual Analog Scale (VAS) pain scale, Neck Disability Index (NDI), and Treatment Intensity Gradient Test (TIGT) scales were evaluated as were static and dynamic radiographs. Significant improvents in all scales were seen postoperatively. One device migration of 4 millimeters was seen at 3 months and observed. Eighty percent of patients had a good or excellent result at one week, improving to ninety percent of patients being judged to have a good or excellent result by one month (Odom's criteria), which then remained at ninety percent at 3 months. PMID- 15541684 TI - Concepts in nuclear replacement. AB - This article describes the pathogenesis of early nuclear failure and current philosophical approaches to artificial, biological, and bioartificial nucleus replacement. Necessary in this review is an overview of current approaches to nucleus replacement with prosthetics and various gel materials. PMID- 15541685 TI - New developments in nucleus pulposus replacement technology. AB - Attempts to alleviate the pain attributed to degeneration of the nucleus pulposus using replacement or reinforcement techniques dating back to the 1950s are reviewed. The various materials and their insertion techniques are discussed as are results available from early clinical experiences. These techniques are in evolution and clinical outcomes will be necessary to establish the efficacy of these approaches. PMID- 15541687 TI - Advances in gene therapy for intervertebral disc degeneration. AB - BACKGROUND CONTEXT: Gene therapy is a growing concept in many fields of medicine, and its potential applications are numerous. With a growing understanding of the molecular and cellular biology of intervertebral disc degeneration, alternatives to current treatment options are under investigation. Gene therapy offers an exciting new direction in the treatment of intervertebral disc degeneration, and potential targets of genetic alteration are being explored. PURPOSE: To describe and update the recent advances in research on gene therapy for the treatment of intervertebral disc degeneration. STUDY DESIGN/SETTING: Review of current research for the application of gene therapy as potential treatment for intervertebral disc degeneration. METHODS: Literature review. RESULTS: There is a growing body of research pertaining to the use of gene therapy as an adjunct or alternative to the current treatment options for intervertebral disc degeneration. In vitro studies have demonstrated that transfer of cDNA encoding growth factors to intervertebral disc cells can favorably modify their metabolic and biological functions. Additionally, initial in vivo studies have demonstrated successful transduction of growth factors to the intervertebral disc with confirmed upregulation of extracellular matrix synthesis. Investigators continue to explore the potential of gene therapy with several factors for the treatment of intervertebral disc degeneration. CONCLUSIONS: The potential of gene therapy to alter the course of intervertebral disc degeneration holds much clinical promise and continues to stimulate further investigations. PMID- 15541686 TI - Growth factors and the intervertebral disc. AB - BACKGROUND CONTEXT: The concept of biologically regenerating the intervertebral disc (IVD) using growth factor injection was first proposed in early 1990. Since that time, the technology to produce recombinant proteins, including growth factors, on an industrial scale has been developed. Currently, the effects of recombinant growth factors on IVD cells are less well known than those on articular cartilage chondrocytes. STUDY DESIGN: Literature review. PURPOSE: The purpose of this review is to summarize the most recent findings of the effects of growth factors on the IVD and, further, to discuss trends in the biological repair of the degenerated intervertebral disc. METHODS: A literature search was conducted using available databases, such as the National Library of Medicine, as well as data presented at scientific conferences held in the past 2 years, primarily in the United States. RESULTS: Herein, we report increasing evidence for the biological regeneration of the IVD. The effect of growth factors on the metabolism of IVD cells or tissues was studied using in vitro and in vivo approaches. In addition to using recombinant growth factor proteins, the gene transfer technique was also applied to study the effect of growth factors on the IVD. For in vivo studies, the availability of recombinant proteins and the use of gene transfer techniques accelerated progress in this field of research. CONCLUSIONS: The results from these in vitro and in vivo studies clearly suggest the potential usefulness of recombinant growth factors as therapeutic drugs or as medical devices. PMID- 15541690 TI - Disc bulge bubble: spine economics 101. PMID- 15541688 TI - Stem cell regeneration of the nucleus pulposus. AB - Low back pain due to disc degeneration is one of the largest health problems faced in this nation when judged by lost work time and direct as well as indirect costs. Many experimental methods are being explored to treat or to reverse the effect of disc degeneration. This article reviews the strategy of a tissue engineering approach to disc regeneration. Mesenchymal stem cells (MSC) such as those obtained from marrow stroma when exposed to the appropriate microenvironment (hypoxia, growth factor, three dimensional culture) differentiate into nucleus pulposus-like cells. These then may be candidates for transplantation and nucleus repopulation. While the work is in its infancy, there is significant optimism that next steps will lead to organ culture models of disc degeneration and regeneration, and ultimately to in vivo rescue of degenerating discs with stem cell treatment. PMID- 15541691 TI - In vitro measurement of pressure in intervertebral discs and annulus fibrosus with and without annular tears during discography. AB - BACKGROUND CONTEXT: Discogram studies have shown that pain reproduction correlates with the extent of annular disruption. However, it has not been assessed if pressure changes in the annulus fibrosus vary incrementally with intradiscal pressure. PURPOSE: To determine the relationship between intradiscal pressure and outer annular pressure during discography in intervertebral discs with and without annular tears. STUDY DESIGN: Intradiscal and periannular pressures were measured simultaneously in vitro during intradiscal injection in porcine cadaver spines. METHODS: Twenty fresh porcine cadaver lumbar spines with intervening discs were tested. Intradiscal and periannular pressures were measured simultaneously using two pressure sensors during intradiscal contrast injection. The tip of a 25-gauge needle connected with a pressure manometer was placed in the center of the nucleus pulposus. A second pressure transducer was located at the outer third of the annulus fibrosus. Needle and transducer locations were confirmed by fluoroscopy. To compare the intact and torn annulus fibrosus, annular disruptions were created with a 20-gauge needle and confirmed by fluoroscopy. RESULTS: During intradiscal injections of discs with an intact annulus, annulus fibrosus pressure remained low and a sharp increase in intradiscal pressure was observed. Significantly higher pressures were noted in the outer annulus in discs with annular tears (p<.01). Mean pressures at the central nucleus pulposus, the outer third of intact annuli and torn annuli were 93.4+/-40.9 psi, 14.8+/-1.9 psi and 85.7+/-24.8 psi, respectively. CONCLUSIONS: Volumetric injection of intervertebral discs with a torn annulus fibrosis during discography may increase intra-annular pressure similar to the increase in pressure that may occur during spinal loading activities. This effect may not occur in discs with an intact annulus fibrosus. PMID- 15541692 TI - The effectiveness of extrication collars tested during the execution of spine board transfer techniques. AB - BACKGROUND CONTEXT: In the prehospital stages of emergency care, cervical collars are (supposedly) used to aid rescuers in maintaining in-line stabilization of the spinal column as patients with potential or actual injuries are shifted onto a spine board to achieve full spinal immobilization. Unfortunately, not a single study has examined the effectiveness of cervical collars to control motion during the execution of spine-board transfer techniques. PURPOSE: To evaluate the controlling effect of three cervical collars during the execution of spine-board transfer techniques. STUDY DESIGN: This was a repeated measures investigation in which a cadaveric model was used to test the effectiveness of the Ambu (Ambu, Inc., Linthicum, MD), Aspen (Aspen Medical Products, Inc., Long Beach, CA) and Miami J (Jerome Medical, Moorestown, NJ) collars during the execution of the log roll (LR) maneuver and the lift-and-slide (LS) technique. METHODS: Six medical professionals executed the LR and the LS on five cadavers. An electromagnetic tracking device was used to capture angular movements generated at the C5-C6 vertebral segment during the execution of both transfer techniques. The types of motion that were analyzed in this study were flexion-extension, lateral flexion and axial rotation motion. To test the three cervical collars, an experimental lesion (ie, a complete segmental instability) was created at the aforementioned spinal level of the cadavers and sensors from the electromagnetic tracking device were affixed to the specified vertebrae to record the motion generated at the site of the lesion. RESULTS: Statistical tests did not reveal a significant interaction between the independent variables of this study (ie, transfer technique and collar type), lending no support to the notion that there may be a combination of collar and transfer technique that could theoretically offer added protection to the patient. Although there was a decrease in the amount of motion generated in every one of the planes of motion as a result of wearing each of the three collars, none of the changes that emerged proved to be significantly different. A significant difference was noted between the LR and LS techniques when the amount of lateral flexion and axial rotation motion generated with each of the procedures were compared. In both cases, execution of the LR maneuver resulted in significantly more motion. CONCLUSIONS: The data presented here suggest that the collars tested in this study are functionally similar. It is recommended that this study be repeated with a larger sample size. PMID- 15541693 TI - Adjacent segment disease after anterior cervical interbody fusion. AB - BACKGROUND CONTEXT: There have been many follow-up studies on anterior interbody fusion for cervical nerve root and spinal cord compression, and excellent neurological outcomes have been reported. However, postoperative degenerative changes at adjacent discs may lead to the development of new radiculopathy or myelopathy. In the previous reports, the incidence of symptomatic adjacent segment disease has ranged from 7% to 15%. PURPOSE: The present study was undertaken to investigate the incidence of symptomatic adjacent segment disease after anterior cervical interbody fusion (ACIF) and to identify the factors that are related to the development of this disease. STUDY DESIGN/SETTING: This is a retrospective cohort study. PATIENT SAMPLE: A total of 112 patients were followed up clinically and radiologically for more than 2 years. OUTCOME MEASURES: Follow up evaluation was primarily by means of clinical visits. The postoperative course of any symptoms, the findings of neurological examination and serial follow-up radiographs were performed in all patients. METHODS: The diagnosis of symptomatic adjacent segment disease was based on the presence of new radiculopathy or myelopathy symptoms referable to an adjacent level, and the presence of a compressive lesion at an adjacent level by magnetic resonance imaging or myelography. We evaluated the correlation between the incidence of symptomatic adjacent segment disease and the following clinical parameters (age at operation, sex, number of the levels fused) and radiological parameters (preoperative cervical spine alignment, preoperative range of motion of C2-C7 cervical spine, anteroposterior spinal canal diameter, preoperative existence of an adjacent segment degeneration on plain radiograph, myelography and magnetic resonance imaging [MRI]). RESULTS: Symptomatic adjacent segment disease developed in 19 of 112 patients (19%) followed. A Kaplan-Meier survival analysis was performed in order to follow the disease-free survival of the entire series of patients. The disease-free survival rates were 89% at 5 years, 84% at 10 years and 67% at 17 years. The incidences of indentation of dura matter on preoperative myelography or disc protrusion on MRI at the adjacent level were significantly higher in disease cases (p=.0087, .0299, respectively; chi-squared test). However, the other parameters did not show a statistically significant difference. There were seven cases (37%) who had failure of nonoperative treatment and additional operations were performed. CONCLUSIONS: The incidence of symptomatic adjacent segment disease after ACIF was higher when preoperative myelography or MRI revealed asymptomatic disc degeneration at that level regardless of the number of the levels fused, preoperative alignment, spinal canal diameter or fusion alignment. PMID- 15541694 TI - Comparison of clinical and radiographic outcome in instrumented anterior cervical discectomy and fusion with or without direct uncovertebral joint decompression. AB - BACKGROUND CONTEXT: Anterior cervical discectomy and fusion (ACDF) is an established procedure for the operative treatment of cervical disc disease in patients with radiculopathy resulting from impingement from uncovertebral joint osteophytes. Studies demonstrate that direct decompression of the lesion provides good result. However, known complications include vertebral artery injury, dural tears, nerve root injury, loss of biomechanical stability and increased operative time. Other studies suggest that disc space distraction may play an important role by indirectly decompressing neural elements. Therefore, if equivalent functional outcomes can be achieved without sacrificing the uncovertebral joint, then potential morbidity and mortality could be decreased. PURPOSE: To assess and compare clinical and radiographic outcomes of patients with neck pain and cervical radiculopathy who underwent instrumented ACDF with or without direct uncovertebral joint decompression. STUDY DESIGN/SETTING: Retrospective clinical chart and radiographic review to assess clinical outcome and graft fusion in 109 patients who underwent one- or two-level ACDF with rigid anterior plate fixation. PATIENT SAMPLE: Radiographs and clinical charts for 109 patients (mean, 46 years; range, 27 to 83) who underwent ACDF with rigid anterior plate fixation were retrospectively reviewed at a single institution. Patients with radiculopathy resulting from herniated disc, spondylosis or a combination of both refractory to conservative treatment underwent surgery using a standard Smith-Robinson left sided approach. Seventy-one patients who received direct uncovertebral joint decompression (Group 1) were compared with 38 patients without direct decompression but indirect decompression by disc space distraction (Group 2). In Group 1, 37 one-level and 34 two-level ACDFs were performed. In Group 2, 11 and 27 were one-level and two-level ACDFs, respectively. Smoking and work-related injuries involved 26.7% and 38.0% of Group 1 and 28.9% and 28.9% of Group 2, respectively. Autologous iliac crest grafts were used in 51 patients, whereas 58 patients received allograft. OUTCOME MEASURES: Independent blinded analyses of plain lateral neutral, flexion and extension radiographs were conducted to assess fusion, evaluate graft and plate and screw integrity (mean, 12 months). Clinical outcomes were reported as excellent, good, fair or poor (mean, 23 months) based on Odom's criteria. METHODS: Postoperative clinical outcome and radiographic studies of graft and instrument integrity were assessed in 71 patients undergoing ACDF with uncovertebral joint decompression and 38 patients without uncovertebral joint decompression, but with indirect decompression through disc space distraction. RESULTS: Fusion occurred in 95.8% of Group 1 and 100% of Group 2. In Group 1, 26.8% of the patients reported excellent results, 57.7% reported good results, 12.7% reported fair results and 2.8% reported poor results. In Group 2, 23.7% of the patients reported excellent results, 60.5% reported good results and 15.8% reported fair results. All nonunions reported good outcomes. Postoperative respiratory distress developed in one patient and dysphagia developed in another both from Group 1. No other complications were noted. The presence or absence of direct uncovertebral joint decompression and clinical outcome was not statistically significant (p>.05). The use of graft-type, operative level, presence of smoking and work-related injury in relation to clinical outcome was not found to be significant (p>.05). CONCLUSION: Good to excellent results were obtained in 84.5% and 84.2% of patients for Groups 1 and 2, respectively. Indirect foraminal decompression through distraction remains somewhat controversial during ACDF. However, sacrificing the uncovertebral joint can increase operative time and potentially increase complication rates. This study demonstrates that ACDF with or without direct uncovertebral joint decompression can provide good clinical results for neck pain with cervical radiculopathy. Therefore, routine direct uncovertebral joint decompression should not be undertaken during ACDF. PMID- 15541695 TI - Does rigid instrumentation increase the fusion rate in one-level anterior cervical discectomy and fusion? AB - BACKGROUND CONTEXT: Although plate fixation enhances the fusion rate in multilevel anterior cervical discectomy and fusion (ACDF), debate exists regarding the efficacy of nonplating to rigid plate fixation in one-level ACDF. PURPOSE: To determine the efficacy of nonplating to rigid plate fixation in regards to fusion rate and clinical outcome in patients undergoing one-level ACDF with autograft. STUDY DESIGN: A review of 69 consecutive patients who underwent one-level ACDF with autograft and with or without rigid anterior cervical plate fixation. PATIENT SAMPLE: Sixty-nine patients who underwent one-level ACDF (mean age, 45 years) were evaluated for radiographic evidence of fusion (mean, 14 months) and for clinical outcome. All patients received tricortical iliac crest autografts. Disc space distraction was 2 mm, the grafts were inserted with the cortical surface positioned anteriorly, and each graft was countersunk 2 mm from the anterior vertebral border. Thirty-eight patients underwent nonplated ACDF and 31 patients underwent plated ACDF. Eighteen Orion (Sofamor-Danek, Memphis, TN), eight Atlantis (Sofamor-Danek) and five PEAK polyaxial (Depuy-Acromed, Rayham, MA) anterior cervical plating systems were used. Rigid plate fixation was used in all patients with instrumentation. Postoperatively, hard collars were worn 6 to 8 weeks in nonplated patients and soft collars were worn for 3 to 4 weeks in plated patients. Twenty-four patients were smokers (54.2% nonplating; 45.8% plating) and work-related injuries entailed 23 patients (47.8% nonplating; 52.2% plating). OUTCOME MEASURES: Fusion was assessed based on last follow-up of lateral neutral, flexion and extension radiographs. Radiographs were evaluated blindly to assess fusion and instrumentation integrity between nonplated and plated patients. Clinical outcomes were assessed with the Cervical Spine Outcomes Questionnaire and also assessed on last follow-up as excellent, good, fair or poor based on Odom's criteria. METHODS: Fusion rate and postoperative clinical outcome were assessed in 69 patients who underwent one-level ACDF with autograft and with or without rigid anterior plate fixation. Additional risk factors were also analyzed. Statistical significance was established at p<.05. RESULTS: Sixty-six patients (95.7%) achieved a solid fusion (100% nonplated; 90.3% plated). Nonunions occurred in three patients (1 smoker; 2 nonsmokers) with Orion instrumentation. Slight screw penetration into the involved and uninvolved interbody spaces occurred in one patient who was a nonsmoker and did not achieve fusion. One superficial cervical wound infection was noted in a nonplated patient. No other intraoperative or postoperative complications were noted. No statistically significant difference was noted between nonplating to rigid plating upon fusion rate (p>.05). All nonunions occurred at the C5-C6 level. Mean estimated intraoperative blood loss was significantly greater in plated patients (p=.043). Revision surgery involved 9.7% of the plated patients, whereas none of the nonplated patients required reoperation. Postoperative clinical outcome was assessed in all patients (mean, 21 months). Excellent results were noted in 18.8%, good results in 72.5% and fair results in 8.7% of the patients. Nonunion patients reported satisfactory clinical outcome. No statistical significance was noted between clinical outcome of fused and nonfused patients, the presence of a work-related injury and the use of plating (p>.05). Demographics and history of smoking were not factors influencing fusion or clinical outcome in this series (p>.05). The effect on fusion by various plate types could not be discerned from this study. CONCLUSION: A 100% and 90.3% fusion rate was obtained for one-level nonplated and plated ACDF procedures with autograft, respectively. The effects of smoking or level of fusion could not be discerned from these one-level cases. Excellent and good clinical outcome results were obtained for 91.3%. Nonplating or rigid plate fixation for ACDF in properly selected patients to treat radiculopathy with or without myelopathy has a high fusion rate and yields a satisfactory clinical outcome. Although controversy exists as to the efficacy of rigid plate fixation in one-level ACDF, solid bone fusion can be adequately obtained without plate fixation and instrumentation-related complications can be avoided. In line with the literature, plate fixation should be reserved for patients unwilling or unable to wear a hard orthosis postoperatively for an extended period of time or for those patients who seek a quicker return to normal activities. Proper patient selection, meticulous operative technique and postoperative care is essential to promote optimal graft-host incorporation. PMID- 15541696 TI - Minimally invasive anterior lumbar interbody fusion followed by percutaneous pedicle screw fixation for isthmic spondylolisthesis. AB - BACKGROUND CONTEXT: Most surgeons have thought that posterior decompression is necessary to treat isthmic spondylolisthesis with leg pain. However, the surgical procedure not only requires wide muscle dissection but can also lead to spinal instability. The authors' treatment concept for isthmic spondylolisthesis is one stage anterior reduction and posterior stabilization with minimally invasive surgical procedure without touching the spinal thecal sac and nerve. PURPOSE: To investigate a new surgical concept of minimally invasive anterior-posterior fusion without posterior decompression for the treatment of isthmic spondylolisthesis with leg pain. STUDY DESIGN: This is a retrospective study of 73 patients with isthmic spondylolisthesis who underwent minimally invasive anterior lumbar interbody fusion (mini-ALIF) followed by percutaneous pedicle screw fixation (PF) between October 2000 and February 2002. PATIENT SAMPLE: A total of 73 patients with low-grade isthmic spondylolisthesis (46 with Grade 1 and 27 with Grade 2) who underwent mini-ALIF followed by percutaneous PF were retrospectively analyzed. There were 20 men and 53 women, with a mean age of 50.6 (range, 19 to 77) years. All patients had low back pain and referred or radicular leg pain or neurogenic intermittent claudication in walking or standing. Average duration of symptoms was 26 (range, 3 to 120) months. OUTCOME MEASURES: The clinical outcome was graded according to the modified Macnab criteria. METHODS: The authors retrospectively reviewed clinical and radiological data of 73 patients who had isthmic spondylolisthesis. All patients underwent mini-ALIF and percutaneous PF on the same day between October 2000 and February 2002. The mean follow-up period was 16 months (range, 12 to 26). RESULTS: The mean operating time, blood loss and hospital stay were 210 minutes, 135 ml and 4.1 days, respectively. No blood transfusion was necessary. Clinical outcome was excellent in 26 patients (35.6%), good in 43 (58.9%), fair in 3 (4.1%) and poor in 1 (1.4%). The fusion rate was 97.3% (71 of 73). There were 6 cases (8.2%) of mini ALIF complications and 6 (8.2%) of percutaneous PF complications. There were no postoperative neurologic deficits. CONCLUSIONS: Mini-ALIF followed by percutaneous PF is an efficacious alternative for low-grade isthmic spondylolisthesis, and posterior decompression is not necessary to relieve leg symptoms. This minimally invasive combined procedure offers many advantages, such as preservation of posterior arch, no nerve retraction, less blood loss, excellent cosmetic results, high fusion rate and early discharge. PMID- 15541697 TI - Prophylaxis of C5 palsy after cervical expansive laminoplasty by bilateral partial foraminotomy. AB - BACKGROUND CONTEXT: It is known that postoperative motor palsy at the C5 level occurs with anterior decompression or posterior decompression and has a relatively good prognosis, but the pathogenesis and possible prophylactic measures of the palsy remain unknown. PURPOSE: The purpose of this study was to evaluate the effectiveness of bilateral partial foraminotomy for preventing C5 palsy from occurring after cervical decompression surgery. STUDY DESIGN: A retrospective review was performed concerning the risk factors of the C5 palsy based on the preoperative clinical findings. To investigate the prophylactic effect of the partial foraminotomy, we examined a difference of an incidence of the C5 palsy by performing concurrent partial foraminotomy with expansive laminoplasty. PATIENT SAMPLE: A total of 305 cases of cervical expansive laminoplasty performed for spondylotic myelopathy or ossification of the posterior longitudinal ligament were reviewed. METHODS: We analyzed 305 cases of cervical expansive laminoplasty to investigate the preoperative risk factors that may cause postoperative C5 palsy. To clarify the relationship of the foraminotomy and development of the C5 palsy, we examined 230 patients in whom foraminotomy could be confirmed by operative records. RESULTS: Of the 305 patients, postoperative C5 palsy occurred in 13 patients (4.3%): 10 patients had radicular pain (77%), and 8 patients had sensory disturbances (62%). We assessed all neurological findings and X-ray, computed tomography and electromyographic findings, but no statistical differences were found in any of the preoperative clinical findings relative to the occurrence of postoperative C5 palsy. For the open side, 108 cases underwent foraminotomy and 122 cases did not, whereas on the hinge side, 54 cases received foraminotomy and 176 cases did not. In order to investigate the prophylactic effect of foraminotomy, we totaled the open side and hinge side, and calculated the number of bone gutters: 162 gutters had concurrent foraminotomy and 298 gutters did not. Postoperatively, C5 palsy occurred in 1 gutter (0.6%) in the former group and in 12 gutters (4.0%) in the latter group (p<.05, Fisher's direct method). CONCLUSIONS: There were no specific risk factors among the preoperative clinical findings related to C5 palsy. Bilateral partial foraminotomy was effective for preventing C5 palsy. PMID- 15541698 TI - Electromyographic and kinematic exploration of whiplash-type rear impacts: effect of left offset impact. AB - BACKGROUND CONTEXT: Although there are some volunteer collision studies reporting the effects of rear impacts on head and neck kinematics, there are few studies detailing the cervical muscle electromyogram response. Moreover, the effect of a rear impact offset to the left on the resultant muscle responses is unknown. PURPOSE: The purpose of this study was to determine the response of the cervical muscles to increasing low-velocity rear impacts offset by 45 degrees to the subject's left, and to compare the quantitative effects of expected and unexpected impact. STUDY DESIGN/SETTING: Nine healthy volunteers were subjected to rear impacts, offset by 45 degrees to the subject's left, of 4.6, 8.7, 11.0 and 14.5 m/s2 acceleration, at two levels of expectation: expected and unexpected. METHODS: Bilateral electromyograms of the sternocleidomastoids, trapezii and splenii capitis were recorded. Triaxial accelerometers recorded the acceleration of the chair, torso at the shoulder level and head of the participant. RESULTS: At an acceleration of 14.5 m/s2, the left sternocleidomastoid generated 71% and the right sternocleidomastoid 82% of their maximal voluntary contraction electromyogram in the unexpected impact conditions. Under these conditions, the right splenius capitis (contralateral to the left offset rear impact) also generated 71% of its maximal voluntary contraction, whereas the left splenius capitis generated only 20% of this variable. The trapezii generated only 25% of their maximal voluntary contraction. Subjects exhibited lower levels of their maximal voluntary contraction electromyogram when the impact was expected. Electromyographic variables were significantly affected by the levels of acceleration and expectation (p<.001). The time to onset and time to peak electromyogram for all muscles progressively decreased with increasing levels of acceleration, in the unexpected condition. The kinetic variables and the electromyographic variables regressed significantly on the acceleration (p<.001). In response to rear impacts offset to the subject's left, muscle responses were greater with higher levels of acceleration, greater with unexpected impact conditions and greatest for both sternocleidomastoids and for the splenius capitis muscle contralateral to the side of impact. CONCLUSIONS: Because the muscular component of the head-neck complex plays a role in the abatement of impact at higher acceleration levels, they are likely a primary site of injury in the whiplash phenomenon in rear collisions. More specifically, when a rear impact is offset to the subject's left, it results in not only increased electromyographic generation in both sternocleidomastoids, but the splenius capitis contralateral to the direction of impact offset also bears part of the force of the neck perturbation. Expecting or being aware of imminent impact also plays a role in reducing muscle responses in low-velocity offset rear impacts. PMID- 15541702 TI - Experimental study of paraplegia caused by spinal tumors: an animal model of spinal tumors created by transplantation of VX2 carcinoma. AB - BACKGROUND: Little is known about the mechanism of the paraplegia produced by metastatic spinal tumors, although the quality of life of patients with paraplegia caused by malignant tumors depends to a great extent on treatment of the paraplegia. We previously established an experimental model of malignant femoral tumors by transplanting VX2 carcinoma into the femur, and we have reported the efficacy of chemical embolic therapy, radiotherapy and hyperthermal therapy in treating the tumors in the model. PURPOSE: The purpose of the present study was to establish an animal model of metastatic spinal tumors by transplanting VX2 carcinoma into a pedicle and to analyze the mechanism of the paraplegia produced by metastatic spinal tumors. STUDY DESIGN: An experimental study. METHODS: VX2 carcinoma that had been successively transplanted to the thigh muscles of Japanese White rabbits in the Department of Orthopaedic Surgery of Kyorin University School of Medicine was used. The third lumbar vertebra of 37 healthy rabbits was exposed, and a piece of VX2 carcinoma was transplanted into a hole created in the pedicle with an air drill after transplantation. The animals were examined every other day. As soon as paraplegia developed, the animals were sacrificed, and the axial localization of the tumor was evaluated based on the surgical staging of the spinal tumor (SSST classification by Tomita). Roentgenographic, histological and angiographic studies were also performed. RESULTS: Twenty-two rabbits (60%) had paraplegia at the time of the final examination. Seventeen of them had complete paraplegia, and the other five had incomplete paraplegia. The average interval until incomplete paraplegia developed was 18.4+/-4.3 days, and the average interval until complete paraplegia developed was 30.0+/-4.3 days. The average proportion of the diameter of the spinal canal occupied by the tumor was 37.2+/-4.9% in the incomplete cases, and 70.6% in the complete cases. The radiographs revealed an osteolytic area in the pedicle and posterior border of the third lumbar vertebra in the rabbits with complete paraplegia, but no osteolytic areas were detected in the rabbits with incomplete paraplegia. According to the SSST classification, the lesions of all of the rabbits with paraplegia were classified as stage IV. Pathological studies revealed that the spinal cord generally exhibited degenerative change, especially at the site of tumor compression. No tumor cells had invaded the spinal cord. Microangiography showed a marked reduction in blood vessels in the gray matter of the spinal cord in the complete paraplegia in comparison with incomplete paraplegia. CONCLUSION: Our method succeeded in establishing a model of metastatic spinal tumors, because the natural history of the disease, including the anatomical location of the tumors and the imaging findings, was similar to the natural history in clinical cases. This model was useful for elucidating the pathogenetic mechanism of the paraplegia caused by metastatic spinal tumors. PMID- 15541701 TI - The effects of doxorubicin (adriamycin) on spinal fusion: an experimental model of posterolateral lumbar spinal arthrodesis. AB - BACKGROUND CONTEXT: Malignant spinal lesions may require surgical excision and segmental stabilization. The decision to perform a concomitant fusion procedure is influenced in part by the need for adjunctive chemotherapy as well as the patient's anticipated survival. Although some evidence exists that suggests that chemotherapy may inhibit bony healing, no information exists regarding the effect of chemotherapy on spinal fusion healing. PURPOSE: To determine the effect of a frequently used chemotherapeutic agent, doxorubicin, on posterolateral spinal fusion rates. STUDY DESIGN/SETTING: Prospective animal model of posterolateral lumbar fusion. OUTCOME MEASURES: Determination of spinal fusion by manual palpation of excised spines. Plain radiographic evaluation of denuded spines to evaluate intertransverse bone formation. METHODS: Thirty-two New Zealand White rabbits underwent posterior intertransverse process fusion at L5-L6 with the use of iliac autograft bone. Rabbits randomly received either intravenous doxorubicin (2.5 mg/kg) by means of the central vein of the ear at the time of surgery (16 animals) or no treatment (16 animals; the control group). The animals were euthanized at 5 weeks, and the lumbar spines were excised. Spine fusion was assessed by manually palpating (by observers blinded to the treatment group) at the level of arthrodesis, and at the adjacent levels proximal and distal. This provided similar information to surgical fusion assessment by palpation in humans. Fusion was defined as the absence of palpable motion. Posteroanterior radiographs of the excised spines were graded in a blinded fashion using a five point scoring system (0 to 4) devised to describe the amount of bone observed between the L5-L6 transverse processes. Power analysis conducted before initiation of the study indicated that an allocation of 16 animals to each group would permit detection of at least a 20% difference in fusion rates with statistical significance at p=.05. RESULTS: Eleven of the 16 spines (69%) in the control group and 6 of the 16 spines (38%) in the doxorubicin group fused. This difference was statistically significant (=.038). There was no significant correlation (p>.05) between the radiographic grade of bone formation (0 to 4) and fusion as determined by palpation. There were four wound infections in the control group and four in the doxorubicin group. However, solid fusions were palpated in three of these four spines in both the control and treatment groups. CONCLUSIONS: No significant differences in wound complications were noted with doxorubicin administration. A single dose of doxorubicin administered intravenously at the time of surgery appears to play a significant inhibitory role in the process of spinal fusion. If similar effects occur in humans, these data suggest that doxorubicin may be harmful to bone healing in a spine fusion if given during the perioperative period. Further investigation will be necessary to determine the effect of time to aid at determining whether doxorubicin administered several weeks pre- or postoperatively results in improved fusion rate, and whether bone morphogenetic proteins can overcome these inhibitory effects. PMID- 15541703 TI - The Lumbar I/F Cage for posterior lumbar interbody fusion with the variable screw placement system: 10-year results of a Food and Drug Administration clinical trial. AB - BACKGROUND CONTEXT: The Lumbar I/F Cage is a carbon fiber reinforced polymer (CFRP) device designed to separate the mechanical and device functions of interbody fusion. A Investigational Device Exemption (IDE) clinical study of the CFRP cage was conducted during an enrollment period from 1991 to 1993. Based on the 2-year results of this study, the cage was approved by the US Food and Drug Administration (FDA) in February 1999. Since then, the Lumbar I/F Cage device has become widely used in the United States. PURPOSE: This study was designed to determine the long-term results of patients who received this device during the 1991-1993 enrollments. STUDY DESIGN/SETTING: Investigators from the original study were asked to evaluate their original patients according to FDA-reviewed case report forms. Although many of the centers were unable to provide significant follow-up, two centers that enrolled almost half of the original study group provided reports on a high percentage of their original patients. This paper reviews the results in those patients. PATIENT SAMPLE: Inclusion criteria included patients with degenerative disc disease who had at least one failed lumbar discectomy or decompression procedure at one or more levels. OUTCOME MEASURES: Clinical success was defined by a modified Prolo score evaluating pain, function, medication usage and economic status. Fusion success, determined by evaluation of plain radiographs, was defined by continuous bone bridging the fusion area with no lucencies. Flexion-extension X-rays were done on patients who had previous removal of pedicle screw implants. Any motion on flexion-extension films indicated pseudarthrosis. METHODS: All patients were contacted at their last known address. Internet search services were used to locate additional patients. Thirty-three of 43 eligible patients (77%) were evaluated, including 31 patients who reported for examination and X-ray and 2 additional patients by telephone survey and written questionnaire. RESULTS: Clinical success was achieved in 32 of 37 patients (86.5%) at 24 months and in 29 of 33 patients (87.8%) at 10 years. This included 61% excellent, 27% good, and 12% fair results. Fusion success was reported in 37 of 37 patients (100%) at 24 months and in 29 of 30 patients (96.7%) at 10 years. Patient satisfaction was reported in 31 of 33 (93.9%). Further lumbar surgery was done in 23 patients: in 18 patients for elective removal of pedicle screws and in 5 patients to extend the fusion to adjacent levels. Adjacent segment degeneration occurred in 61% of patients but was clinically significant in only 20%. Smokers had equal clinical and fusion success with nonsmokers at 24 months and 10 years and had adjacent segment degeneration in 37%, a rate significantly lower than nonsmokers at 87%. CONCLUSIONS: The high rate of clinical success, fusion success, and patient satisfaction at 24 months was maintained at 10-year follow-up. Adjacent segment degeneration was common but was usually not clinically significant. PMID- 15541704 TI - Rheumatoid arthritis: evaluation and surgical management of the cervical spine. AB - BACKGROUND CONTEXT: Rheumatoid arthritis is a debilitating polyarthropathic degenerative condition. Eighty-six percent of patients with rheumatoid arthritis have cervical spine involvement. Often these lesions are clinically asymptomatic or symptoms are erroneously attributed to peripheral manifestation of the patient's rheumatoid disease. Because these lesions are common and missed diagnosis can result in death, early recognition is vital. PURPOSE: The purpose of this literature review is to identify common lesions present in the rheumatoid neck and review diagnostic methods as well as treatment options for those requiring surgical intervention. STUDY DESIGN: A review of the English medical literature with focus on more recent studies on the presentation, diagnosis, management, surgical treatment and clinical outcomes of rheumatoid arthritis of the cervical spine. METHODS: A comprehensive literature review of the English medical literature obtained through Medline up to November 2003 was performed identifying relevant and more recent articles that addressed the presentation, evaluation, surgical management and outcomes of rheumatoid patients with cervical spine involvement. RESULTS: If left untreated, a large percentage of rheumatoid patients with cervical spine involvement progress toward complex instability patterns resulting in significant morbidity and mortality. Once myelopathy occurs, prognosis for neurologic recovery and long-term survival is poor. In properly selected patients, anterior and/or posterior cervical procedures can prevent neurologic injuries and preserve remaining function. CONCLUSION: Cervical spine involvement in the rheumatoid patient is common and progressive. Early diagnosis and treatment is imperative; however, surgical intervention should be considered carefully because associated morbidity and mortality is high. PMID- 15541705 TI - Unnecessary cervical disc surgery for a malingering patient. PMID- 15541707 TI - Cognitive reserve and the neurobiology of cognitive aging. AB - A hypothetical construct of "cognitive reserve" is widely used to explain how, in the face of neurodegenerative changes that are similar in nature and extent, individuals vary considerably in the severity of cognitive aging and clinical dementia. Intelligence, education and occupational level are believed to be major active components of cognitive reserve. Here, we summarize the main features of cognitive aging and their neuropathological correlates. We describe the neurobiology of cognitive aging and conclude that perturbations of neural health attributable to oxidative stress and inflammatory processes alone are insufficient to distinguish cognitive aging from Alzheimer's disease. We introduce the concept of cognitive reserve and illustrate its utility in explaining individual differences in cognitive aging. Structural and functional brain imaging studies suggest plausible neural substrates of cognitive reserve, probably involving processes that support neuroplasticity in the aging brain. The cognitive reserve hypothesis conforms with reported associations between early and mid life lifestyle choices, early education, lifelong dietary habit, leisure pursuits and the retention of late life mental ability. PMID- 15541708 TI - Biophysical alterations of hippocampal pyramidal neurons in learning, ageing and Alzheimer's disease. AB - A series of behavioral, electrophysiological, and molecular biochemical experiments are reviewed indicating that when animals learn hippocampus-dependent tasks, output neurons in the CA1 and CA3 hippocampal subfields show reductions in the slow, post-burst afterhyperpolarization (AHP). The slow AHP is mediated by an apamin-insensitive calcium-activated potassium current. A reduction in the slow AHP makes hippocampal neurons more excitable and facilitates NMDA receptor mediated response and temporal summation. During normal aging and in a mouse model of Alzheimer's disease (AD), the slow AHP is increased, making neurons less excitable and making learning more difficult. The subgroup of aging animals that are able to learn demonstrates the capacity to increase neuronal excitability by reducing the size of the slow AHP. Similarly, in a mouse model of AD, mice that are able to learn normally after a genetic alteration have a normal capacity for increasing hippocampal neuron excitability by reducing their slow AHP. We suggest that reduction in the slow AHP is basic to learning in young and aging animals. Inability to modulate the slow AHP contributes to learning deficits that occur during aging and early stages of AD. PMID- 15541709 TI - Regulation of late-phase LTP and long-term memory in normal and aging hippocampus: role of secreted proteins tPA and BDNF. AB - Long-lasting forms of memory are generally believed to be mediated by protein synthesis-dependent, late-phase long-term potentiation (L-LTP). L-LTP exhibits at least two distinctive characteristics compared with early phase LTP (E-LTP): synaptic growth and requirement of gene transcription and new protein synthesis. In this review, we discuss the cellular and molecular mechanisms underlying the structural and functional changes of hippocampal synapses during L-LTP, in the context of long-term memory. We describe experiments that reveal the critical role of cAMP/protein kinase A and MAP kinase pathways, and the downstream transcription factor CREB. Because transcription-dependent long-term changes are input specific, we also discuss the role of "local protein synthesis" and "synaptic tagging" mechanisms that may confer synapse specificity. We then focus on brain-derived neurotrophic factor (BDNF) and tissue plasminogen activator (tPA), two secreted proteins that have been repeatedly implicated in L-LTP. Biochemical and molecular biology experiments indicate that the expression and secretion of both factors are enhanced by strong tetanic stimulation that induces L-LTP as well as by training in hippocampal-dependent memory tasks. Inhibition of either tPA or BDNF by gene knockout and specific inhibitors results in a significant impairments in L-LTP and long-term memory. Further work will be required to address the relationship between BDNF and tPA in various forms of synaptic plasticity, and the mechanisms by which BDNF/tPA achieves synapse specific modulation. Finally, we discuss how the aging process affects L-LTP and long-term memory. PMID- 15541710 TI - Reactive oxygen species and synaptic plasticity in the aging hippocampus. AB - Aging is associated with a general decline in physiological functions including cognitive functions. Given that the hippocampus is known to be critical for certain forms of learning and memory, it is not surprising that a number of neuronal processes in this brain area appear to be particularly vulnerable to the aging process. Long-term potentiation (LTP), a form of synaptic plasticity that has been proposed as a biological substrate for learning and memory, has been used to examine age-related changes in hippocampal synaptic plasticity. A current hypothesis states that oxidative stress contributes to age-related impairment in learning and memory. This is supported by a correlation between age, memory impairment, and the accumulation of oxidative damage to cellular macromolecules. However, it also has been demonstrated that ROS are necessary components of signal transduction cascades during normal physiological processes. This review discusses the evidence supporting the dual role of reactive oxygen species (ROS) as cellular messenger molecules in normal LTP, as well their role as damaging toxic molecules in the age-related impairment of LTP. In addition, we will discuss parallel analyses of LTP and behavioral tests in mice that overexpress antioxidant enzymes and how the role of antioxidant enzymes and ROS in modulating these processes may vary over the lifespan of an animal. PMID- 15541711 TI - A neural signaling triumvirate that influences ageing and age-related disease: insulin/IGF-1, BDNF and serotonin. AB - The ageing process and its associated diseases all involve perturbed energy metabolism, oxidative damage, and an impaired ability of the organism and its cells to cope with adversity. We propose that some specific signaling pathways in the brain may be important determinants of health during ageing. Among such specific signaling modalities are those activated in neurons by insulin-like growth factors (IGFs), brain-derived neurotrophic factor (BDNF) and serotonin. This triumvirate may be particularly important because of their cooperative influence on energy metabolism, food intake, stress responses and cardiovascular function. The health benefits to the periphery and central nervous system of dietary restriction and exercise may be mediated by this triumvirate of signals in the brain. At the molecular level, BDNF, serotonin and IGFs can all stimulate the production of proteins involved in cellular stress adaptation, growth and repair, neurogenesis, learning and memory and cell survival. The importance of this triumvirate is emphasized when it is seen that their general roles in energy metabolism, stress adaptation and disease resistance are conserved among diverse organisms consistent with important roles in the ageing process. PMID- 15541712 TI - Aging and neuronal replacement. AB - Neural stem cells contribute to neurogenesis in both the embryonic and adult brain. However, while adult neural stem cells produce new neurons that populate the olfactory bulb and the granule cell layer of the hippocampus, they do not normally participate in reparative neurogenesis following injury or disease affecting regions distant from the subventricular zone or the dentate gyrus. Here we review differences between neural stem cells found in the embryo and the adult, and describe factors that enhance neuronal output from these cells in vivo. Additionally, we review evidence that neural stem cells can be transplanted into injured regions of the adult brain to enhance compensatory neurogenesis from endogenous precursors. Pre-differentiation of neural stem cells into immature neurons prior to transplantation can also aid in functional recovery following injury or disease. PMID- 15541714 TI - Fas ligand is enriched in the caveolae membrane domains of thymic epithelial cells. AB - Both Fas and Fas ligand (FasL) are expressed in the thymus. Although reports suggest that they are important throughout the thymocyte maturation process their precise role remains elusive. The present paper characterizes the expression of FasL in the thymus and in the TEA3A1 and BT1B functional thymic epithelial cell (TEC) lines. FasL expression by thymus fractions, TEA3A1, and BT1B cells was detected by Northern blot analysis. In TEA3A1 cells, we discovered that FasL protein expression was localized to caveolae membrane domains. This restricted subcellular localization of FasL, together with reports describing the localization of the major histocompatibility complex proteins, the T cell receptor and Fas to caveolae membrane domains, may provide a mechanism for the deletion of thymocytes during negative selection. Finally, using semi quantitative RT-PCR we found that FasL expression by TECs is regulated by glucocorticoids. PMID- 15541713 TI - Chronic nicotine inhibits inflammation and promotes influenza infection. AB - Epidemiological data suggest an association between smoking, respiratory infections, and impaired wound healing. Inflammation is critical in the body's defense against pathogens and in the wound-healing process. Although nicotine is used to treat some inflammatory conditions, the mechanism of this action is largely unknown. To determine how nicotine affects inflammation, rats and mice were exposed to nicotine via miniosmotic pumps, and the inflammatory response to turpentine or influenza virus was assessed. Results showed that while nicotine suppressed the migration of leukocytes to the inflammation/infection site, it increased the influenza titer in the lung. The decreased inflammation correlated with lower chemotaxis/chemokinesis of peripheral blood mononuclear cells (PBMC) toward formyl-methionyl-leucyl-phenylalanine and monocyte chemoattractant protein 1 without affecting the density of their respective receptors. However, nicotine suppressed the chemokine-induced Ca(2+) response in PBMC, indicating impaired chemokine signaling. Thus, because nicotine suppresses leukocyte migration, it might contribute to the delayed wound healing and increased incidence of respiratory infections among smokers. PMID- 15541715 TI - Differences in the expression of LPS-receptors are not responsible for the sex specific immune response after trauma and hemorrhagic shock. AB - Several studies demonstrated a sex-specific cytokine secretion by macrophages following trauma-hemorrhage (T-H) and incubation with lipopolysaccharide A (LPS). Although LPS is known to act via the receptors CD14 and TLR4 on macrophages, it remains unknown whether differences in LPS receptor expression in males and females may be responsible for the gender-specific LPS induced cytokine response following (T-H). To study this, male and proestrus female mice (C3H/HeN) were subjected to trauma (laparotomy) followed by hemorrhage or sham operation. At 2 h thereafter, SMphi and PMphi were harvested and cultured for 2 h. The expression of CD14 and TLR4 was measured by flow cytometry on unstimulated SMphi and PMphi as well as after LPS stimulation. The results indicate that the expression of CD14 and TLR4 on SMphi and PMphi from female and male mice was similar in sham operated animals and after (T-H). Incubation of macrophages with LPS did not alter CD14 and TLR4 expression in the study groups. Thus, the sex specific LPS induced cytokine secretion after (T-H) is not caused by differences in LPS receptor expression on Mphi of male and female mice. PMID- 15541716 TI - Synergy between IL-8 and GM-CSF in reproductive tract epithelial cell secretions promotes enhanced neutrophil chemotaxis. AB - Neutrophils occur in tissues of the female reproductive tract (FRT) under non infected conditions. These cells generally enter tissues under the influence of chemoattractants called chemokines. Primary epithelial cells (EC) from FRT were a potent source of chemokines, IL-8 being the chief neutrophil chemoattractant secreted. Blocking with neutralizing anti-IL-8 showed that IL-8 did not account for all of the chemoattraction observed. A mixture of 25 ng/mL rIL-8 and 1 ng/mL rGM-CSF mediated 2.7-fold more chemotaxis than that expected if the two agents were additive. We then found that GM-CSF was produced by EC in amounts that synergised strongly with IL-8 to enhance chemotaxis. Treatment of uterine EC conditioned medium with saturating doses of anti-IL-8 plus anti-GM-CSF antibodies produced an 84% inhibition of chemotaxis. These findings demonstrate that the majority of neutrophil chemoattractant activity produced by FRT EC results from the synergistic effects of IL-8 and GM-CSF. PMID- 15541717 TI - MHC class II biosynthesis by activated rat CD4+ T cells: development of repression in vitro and modulation by APC-derived signals. AB - This study focused on synthesis of MHC class II glycoproteins (MHCII) by rat CD4(+) T-helper cells. During activation in Con A and IL-2, purified rat splenic CD4(+) T cells expressed abundant surface MHCII together with transcripts for I-A alpha/beta, invariant chain, and the type III and type IV MHC class II transactivator (CIITA). Activated thymic CD8(+)CD4(-) and CD8(+)CD4(+) T cells exhibited essentially the same phenotype. MHCII synthesis by CD4(+) T cells enabled presentation of myelin basic protein (MBP) to antigen-specific responders. T cell expression of MHCII was due to direct biosynthesis rather than adsorption from professional APC; indeed, T cell-mediated expression of MHCII was optimal in the absence of professional APC. Despite periodic reactivation with Con A during 3-4 weeks of culture, CD4(+) T cells repressed MHCII synthesis and reverted to a MHCII(-) phenotype. These short-term lines resembled established lines of MBP-specific T cells in that mitogenic activation elicited extensive blastogenesis without MHCII synthesis. Activation-dependent synthesis of MHCII however was partially restored in lines of mitogen-stimulated T cells when the cultures were reconstituted with irradiated splenic APC. These data indicate that most naive rat CD4(+) T cells exhibit activation-dependent synthesis of MHCII whereas continuously propagated T cells require an APC-derived signal to support MHCII synthesis. PMID- 15541718 TI - Activation and migration of allo-peptide specific TCR transgenic T cells in cardiac allograft rejection. AB - T cells from TCR transgenic mice, expressing receptors specific for an allogeneic MHC class I peptide, were used to track T cell activation and migration in normal adoptive recipients that were subsequently transplanted with heterotopic hearts that were syngeneic except for a transgenic MHC class I antigen. T cells rapidly disappeared from the blood into the lymphoid tissues where they were activated within one day after transplantation. T cells initially formed discrete clusters in the spleen and lymph nodes. After proliferating for 2-4 days in lymphoid tissues, T cells reappeared in the blood and migrated to the heart and the intestines. The T cells underwent another round of proliferation in the heart, but not the intestines, and induced cardiac rejection uniformly on 6 day. PMID- 15541719 TI - Chlamydia trachomatis pulmonary infection induces greater inflammatory pathology in immunoglobulin A deficient mice. AB - Chlamydia trachomatis is an intracellular bacterial pathogen that primarily infects via mucosal surfaces. Using mice with a targeted disruption in IgA gene expression (IgA(-/-) mice), we have studied the contribution of IgA, the principal mucosal antibody isotype, in primary immune defenses against pulmonary C. trachomatis infection. Bacterial burden was comparable between IgA(-/-) and IgA(+/+) animals following C. trachomatis challenge. Serum and pulmonary anti Chlamydia antibody levels were higher in IgA(-/-) animals, with the exception of IgA. Lung sections of challenged IgA(-/-) mice showed more extensive immunopathology than corresponding IgA(+/+) animals. Real-time PCR analysis demonstrated significantly greater IFN-gamma and TGF-beta mRNA expression in IgA( /-) as compared to IgA(+/+) animals. Together, these results suggest that IgA may not be necessary for clearance of primary C. trachomatis infection. However, IgA( /-) mice displayed exaggerated lung histopathology and altered cytokine production, indicating an important role for IgA in regulating C. trachomatis induced pulmonary inflammation and maintenance of mucosal homeostasis. PMID- 15541720 TI - Decreased tumorigenicity of c-Myc-transformed fibroblasts expressing active USF2. AB - USF is a small family of basic helix-loop-helix leucine zipper (bHLH-zip) transcription factors with DNA binding specificities similar to that of the c-Myc oncoprotein. Evidence for a role of USF in growth control includes inhibition of c-Myc-dependent cellular transformation in vitro and loss of USF transcriptional activity in many cancer cell lines. However, a direct effect of USF on the tumorigenicity of an established cell line has never been demonstrated. Here, cell lines derived from rat embryo fibroblasts transformed by c-Ha-Ras and either c-Myc or E1A were used as model system to investigate the tumor suppression ability of USF. Overexpression of USF2 stimulated transcription and inhibited colony formation in c-Myc-transformed, but not E1A-transformed, fibroblasts. Stable clones expressing high USF2 levels were constructed from c-Myc-transformed fibroblasts. In two of these clones, overexpressed USF2 did not activate transcription, and there was no significant change in the transformed phenotype. In contrast, a clone that expressed transcriptionally active USF2 exhibited altered morphology and a strongly decreased ability to proliferate in semisolid medium. The ability of these cells to form tumors in nude mice was also decreased by a factor of more than 30 as compared to the parental cell line or cells overexpressing transcriptionally inactive USF2. Cotransfection assays with USF- or Myc-specific dominant-negative mutants indicated that active USF2 inhibited cellular transformation by preventing transcriptional repression by c-Myc. PMID- 15541721 TI - Activation of p21CIP1/WAF1 gene expression and inhibition of cell proliferation by overexpression of hepatocyte nuclear factor-4alpha. AB - The F9 murine embryonal carcinoma cell line provides an attractive system for studying epithelial differentiation and antiproliferative processes. We have recently established F9 cells expressing doxycycline-inducible hepatocyte nuclear factor (HNF)-4alpha and shown that HNF-4alpha triggers the gene expression of tight-junction molecules, occludin, claudin-6, and claudin-7, as well as formation of functional tight junctions and polarized epithelial morphology (Exp. Cell Res. 286, [2003] 288). Since these events were very similar to those induced by retinoids, we investigated whether HNF-4alpha, like retinoid receptors, was involved in the control of cell proliferation. We herein show that HNF-4alpha up regulates expression of the p21 gene, but not the p15, p16, p18, p19, or p27 gene, in a p53-independent manner, and inhibits cell growth in F9 cells. Similar results were observed in rat lung endothelial cells, in which expression of HNF 4alpha is conditionally induced by doxycycline. Furthermore, we demonstrate, by reporter assay, that HNF-4alpha significantly elevates the transcriptional activity of the p21 promoter. Since, HNF-4alpha is expressed not only in the liver but also in organs containing epithelial cells, such as kidney, intestine, pancreas, and stomach, it might also play critical roles in the regulation of epithelial morphogenesis and proliferation in these organs. PMID- 15541722 TI - A novel alternative spliced Mpv17-like protein isoform localizes in cytosol and is expressed in a kidney- and adult-specific manner. AB - Mpv17-like protein (M-LP) has been identified as a new protein that shows high sequence homology with Mpv17 protein, a peroxisomal membrane protein involved in the development of early onset glomerulosclerosis. We previously showed that the originally identified M-LP isoform, designated M-LPL, is, like Mpv17, localized in peroxisomes, and that transfection with M-LPL up-regulates expression of the manganese superoxide dismutase (SOD2) gene [R. Iida, T. Yasuda, E. Tsubota, H. Takatsuka, M. Masuyama, T. Matsuki, K. Kishi, M-LP, Mpv17-like protein, has a peroxisomal membrane targeting signal comprising a transmembrane domain and a positively charged loop and up-regulates expression of the manganese superoxide dismutase gene. J. Biol. Chem. 278 (2003) 6301-6306.]. We report here the identification of a novel alternative splicing product of the M-LP gene, designated M-LPS. A comparison of the genomic sequence with the cDNA sequences and an analysis of 5'-flanking regions revealed that the two isoforms are generated by alternative usage of two promoters. M-LPS consists of the C-terminal half of M-LPL (90 amino acids) and therefore lacks the peroxisome targeting signal of membrane protein that exists near the N-terminus of M-LPL. Expression of green fluorescent protein-tagged M-LPS in COS-7 cells demonstrated that M-LPS localizes in the cytosol. In mice, M-LPS is expressed exclusively in kidneys after the age of 6 weeks. Moreover, quantitative real-time PCR analysis revealed that transfection with M-LPS up-regulates expression of the SOD2 gene and down regulates expression of the cellular glutathione peroxidase (Gpx1) and plasma glutathione peroxidase (Gpx3) genes. Taken together, these results suggest different functional attributes of the two M-LP isoforms during aging and development. PMID- 15541723 TI - Leukotriene D4 activates distinct G-proteins in intestinal epithelial cells to regulate stress fibre formation and to generate intracellular Ca2+ mobilisation and ERK1/2 activation. AB - We have shown that the pro-inflammatory mediator LTD4, via its G-protein-coupled receptor CysLT1, signals through both pertussis-toxin-sensitive and -insensitive G-proteins to induce various cellular responses. To further characterise the initial step of the different signalling pathways emanating from the CysLT1 receptor, we transfected intestinal epithelial cells, Int 407, with different mini vectors that each express a specific inhibitory peptide directed against a unique alpha subunit of a specific heterotrimeric G-protein. Our results revealed that LTD4-induced stress fibre formation is inhibited approximately 80% by a vector expressing an inhibitory peptide against the pertussis-toxin-insensitive Galpha12-protein in intestinal epithelial Int 407 cells. Control experiments revealed that the LPA-induced stress fibre formation, mediated via the Galpha12 protein in other cell types, was blocked by the same peptide in intestinal Int 407 cells. Furthermore, the CysLT1-receptor-mediated calcium signal and activation of the proliferative ERK1/2 kinase are blocked in cells transfected with a vector expressing an inhibitory peptide against the Galphai3-protein, whereas in cells transfected with an empty ECFP-vector or vectors expressing inhibitory peptides against the Galphai1-2-, Galpha12-, GalphaR-proteins these signals are not significantly affected. Consequently, the CysLT1 receptor has the capacity to activate at least two distinctly different heterotrimeric G-proteins that transduce activation of unique downstream cellular events. PMID- 15541724 TI - Survival and neural differentiation of adult neural stem cells transplanted into the mature inner ear. AB - The cochlear sensory epithelium and spiral ganglion neurons (SGNs) in the adult mammalian inner ear do not regenerate following severe injury. To replace the degenerated SGNs, neural stem cell (NSC) is an attractive alternative for substitution cell therapy. In this study, adult mouse NSCs were transplanted into normal and deafened inner ears of guinea pigs. To more efficiently drive the implanted cells into a neuronal fate, NSCs were also transduced with neurogenin 2 (ngn2) before transplantation. In deafened inner ears and in animals transplanted with ngn2-transduced NSCs, surviving cells expressed the neuronal marker neural class III beta-tubulin. Transplanted cells were found close to the sensory epithelium and adjacent to the SGNs and their peripheral processes. The results illustrate that adult NSCs can survive and differentiate in the injured inner ear. It also demonstrates the feasibility of gene transfer to generate specific progeny for cell replacement therapy in the inner ear. PMID- 15541725 TI - Protocadherin 12 (VE-cadherin 2) is expressed in endothelial, trophoblast, and mesangial cells. AB - Protocadherin 12 protein (PCDH12, VE-cadherin 2) is a cell adhesion molecule that has been isolated from endothelial cells. Here, we have used Northern and Western blots, immunohistology, and flow cytometry to examine the distribution of PCDH12 in mouse tissues. It is an N-glycosylated protein of 150-kDa mass. In the endothelium, PCDH12 immunoreactivity was variable and dependent upon the vascular bed. In both the embryo and embryonic stem cell differentiation system, signals were localized in vasculogenic rather than angiogenic endothelium. In addition, the protein was strongly expressed in a subset of invasive cells of the placenta, which were identified as glycogen-rich trophoblasts. In adult mice, strong PCDH12 signals were observed in mesangial cells of kidney glomeruli whereas expression was not detected in other types of perivascular cells. As opposed to most protocadherins, PCDH12 is not expressed in early embryonic (day 12.5) and adult brains. As a first approach to obtain insight into PCDH12 function, we produced transgenic mice deficient in PCDH12, which were viable and fertile. They did not display any obvious histomorphological defects. We conclude that PCDH12 has a unique expression pattern and that its deficiency does not lead to conspicuous abnormalities. Moreover, PCDH12 is the first specific marker for both glycogen rich trophoblasts and mesangial cells. PMID- 15541726 TI - Association of PAP-1 and Prp3p, the products of causative genes of dominant retinitis pigmentosa, in the tri-snRNP complex. AB - PAP-1 has been identified by us as a Pim-1-binding protein and has recently been implicated as the defective gene in RP9, one type of autosomal dominant retinitis pigmentosa (adRP). We have then shown that PAP-1 plays a role in pre-mRNA splicing. Because four causative genes for adRP, including PAP-1, Prp31, Prp8, and Prp3, encode proteins that function as splicing factors or splicing modulating factors, we investigated the interaction of PAP-1 with Prp3p and Prp31p in this study. The results showed that PAP-1 interacted with Prp3p but not Prp31p in human cells and yeast, and that the basic region of PAP-1 and the C terminal region of Prp3p, regions beside spots found in adRP mutations, were needed for binding. Furthermore, both Prp3p and a part of PAP-1 were found to be components of the U4/U6.U5-tri-snRNP complex, one form of the spliceosome, in Ba/F3 and K562 cells by analysis of sucrose density gradients, suggesting that PAP-1 is weakly associated with the spliceosome. These results also suggest that splicing factors implicated in adRP contribute alone or mutually to proper splicing in the retina and that loss of their functions leads to onset of adRP. PMID- 15541727 TI - Survivin interacts with Smac/DIABLO in ovarian carcinoma cells but is redundant in Smac-mediated apoptosis. AB - Abnormalities in the control and execution of apoptosis are seen in many malignancies, including ovarian carcinoma. Many of these abnormalities involve the mitochondrial pathway of apoptosis, including overexpression of BIR containing inhibitor of apoptosis protein (IAP) family proteins as well as dysregulated apoptosome function. We sought to stimulate the mitochondrial pathway of apoptosis by constructing a recombinant adenovirus encoding mature, processed Smac/DIABLO (Ad CMV tSmac), the second mitochondrial activator of caspases. Transfection of ovarian carcinoma cells with Ad CMV tSmac leads to increasing apoptosis in a dose-dependent manner. By contrast, transfection of IOSE397 immortalized normal ovarian surface epithelial cells does not cause apoptosis. We also show that the processed form of Smac is primarily expressed in the cytosol of ovarian carcinoma cells. Smac co-immunoprecipitates with both survivin and XIAP and stimulates survivin, but not XIAP, down-regulation. This down-regulation does not result from transcriptional changes, as determined by quantitative real-time PCR, but cycloheximide treatment indicates that survivin half-life is reduced from 6 to 2 h, which is secondary to ubiquitination and proteasomal degradation. RNA interference, however, suggests that survivin does not act to inhibit Smac-mediated apoptosis, which is confirmed by cotransfection with the phosphorylation mutant, survivin T34A. Finally, intraperitoneal delivery of Ad CMV tSmac increases median survival of mice bearing human ovarian carcinoma xenografts. We believe that expression of Smac/DIABLO can stimulate the intrinsic pathway of apoptosis in ovarian carcinoma without damaging normal ovarian tissue and therefore has therapeutic potential. PMID- 15541728 TI - Membrane ruffles in cell migration: indicators of inefficient lamellipodia adhesion and compartments of actin filament reorganization. AB - During epithelial cell migration, membrane ruffles can be visualized by phase contrast microscopy as dark waves arising at the leading edge of lamellipodia that move centripetally toward the main cell body. Despite the common use of the term membrane ruffles, their structure, molecular composition, and the mechanisms leading to their formation remained largely unknown. We show here that membrane ruffles differ from the underlying cell lamella by more densely packed bundles of actin filaments that are enriched in the actin cross-linkers filamin and ezrin, pointing to a specific bundling process based on these cross-linkers. The accumulation of phosphorylated, that is, inactivated, cofilin in membrane ruffles suggests that they are compartments of inhibited actin filament turnover. High Rac1 and low RhoA activities were found under conditions of suboptimal integrin ligand interaction correlating with low lamellipodia persistence, inefficient migration, and high ruffling rates. Based on these findings, we define membrane ruffles as distinct compartments of specific composition that form as a consequence of inefficient lamellipodia adhesion. PMID- 15541730 TI - A basic peptide within the juxtamembrane region is required for EGF receptor dimerization. AB - The epidermal growth factor receptor (EGFR) is fundamental for normal cell growth and organ development, but has also been implicated in various pathologies, notably tumors of epithelial origin. We have previously shown that the initial 13 amino acids (P13) within the intracellular juxtamembrane region (R645-R657) are involved in the interaction with calmodulin, thus indicating an important role for this region in EGFR function. Here we show that P13 is required for proper dimerization of the receptor. We expressed either the intracellular domain of EGFR (TKJM) or the intracellular domain lacking P13 (DeltaTKJM) in COS-7 cells that express endogenous EGFR. Only TKJM was immunoprecipitated with an antibody directed against the extracellular part of EGFR, and only TKJM was tyrosine phosphorylated by endogenous EGFR. Using SK-N-MC cells, which do not express endogenous EGFR, that were stably transfected with either wild-type EGFR or recombinant full-length EGFR lacking P13 demonstrated that P13 is required for appropriate receptor dimerization. Furthermore, mutant EGFR lacking P13 failed to be autophosphorylated. P13 is rich in basic amino acids and in silico modeling of the EGFR in conjunction with our results suggests a novel role for the juxtamembrane domain (JM) of EGFR in mediating intracellular dimerization and thus receptor kinase activation and function. PMID- 15541729 TI - Estrogen regulation of trefoil factor 1 expression by estrogen receptor alpha and Sp proteins. AB - Estrogen-responsive genes in human breast cancer cells often have an estrogen response element (ERE) positioned next to an Sp1 binding site. In chromatin immunoprecipitation (ChIP) assays, we investigated the binding of estrogen receptor alpha (ER), Sp1, and Sp3 to the episomal and native estrogen-responsive trefoil factor 1 (TFF1; formerly pS2) promoter in MCF-7 breast cancer cells. Mutation of the Sp site upstream of the ERE reduced estrogen responsiveness and prevented binding of Sp1 and Sp3, but not ER to the episomal promoter. In the absence of estradiol (E2), Sp1, Sp3, histone deacetylase 1 (HDAC), and HDAC2, and low levels of acetylated H3 and H4 are associated with the native promoter, with the histones being engaged in dynamic reversible acetylation. Following E2 addition, levels of ER and acetylated H3 and H4 bound to the native promoter increases. There is clearance of Sp1, but not of Sp3, from the promoter while HDAC1 and HDAC2 remain bound. These data are consistent with a model in which Sp1 or Sp3 aid in recruitment of HDACs and histone acetyltransferases (HATs) to mediate dynamic acetylation of histones associated with the TFF1 promoter, which is in a state of readiness to respond to events occurring following the addition of estrogen. PMID- 15541731 TI - Caspase-9 plays a marginal role in serum starvation-induced apoptosis. AB - Serum withdrawal represents a potent trigger to induce caspase-dependent apoptosis in a series of cell culture models. In rat 423-cells, caspase-8 and caspase-3 were apparently sufficient to initiate and proceed apoptosis without involving the intrinsic amplification loop via caspase-9. To assess the reasons for this inactivity of an otherwise crucial initiator caspase, we examined the ability for apoptosome assembly in 423-cells. Caspase-9 and Apaf-1 were expressed and cytochrome c released from mitochondria upon serum withdrawal. Although functional apoptosomes were assembled successfully in vitro, caspase-9 processing was found essentially refrained during apoptosis in 423-cells. Cell fractionation experiments revealed that sequestration of caspase-9 to cytoskeletal structures in 423-cells contributed to the observed impairment of apoptosome formation. Altogether, these findings provide evidence that serum starvation-induced apoptosis may occur independently of the intrinsic pathway and that caspase-9 sequestration potentially represents a novel biological antiapoptotic strategy. PMID- 15541732 TI - Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. AB - To determine the molecular mechanisms by which N-cadherin transcription is regulated, we cloned and sequenced a 3681-bp of the 5'-flanking region of the human N-cadherin gene. Deletion analysis of the proximal region identified a minimal 318-bp region with strong promoter activity in human osteoblasts. The cryptic promoter is characterized by high GC content and a GA-rich binding core that may bind zing finger transcription factors. Electrophoretic mobility shift assays (EMSA), competition and supershift EMSA revealed that an Sp1/Sp3 binding site acts as a basal regulatory element of the promoter in osteoblasts. Incubation of osteoblast nuclear extracts with -163/-131 wild-type probe containing the GA-rich binding core revealed another specific complex, which was not formed with a -163/-131 probe mutated in the GA repeat. EMSA identified the nuclear factor involved as myeloid zinc finger-1 (MZF1). Mutation analysis showed that Sp1/Sp3 and MZF1 binding sites contribute to basal promoter activity. Cotransfection analyses showed that Sp1 and MZF1 overexpression increases whereas Sp3 antagonizes Sp1-induced N-cadherin promoter activity in osteoblasts. RT-PCR analysis showed that human osteoblastic cells express MZF1 and that Sp1/MZF1 overexpression increased N-cadherin expression. These results indicate that Sp1/Sp3 and MZF1 are important transcription factors regulating N-cadherin promoter activity and expression in osteoblasts. PMID- 15541733 TI - Bufadienolides from Drimia robusta and Urginea epigea (Hyacinthaceae). AB - Two bufadienolides, 6beta-acetoxy-3beta,8beta,14beta-trihydroxy-12-oxobufa 4,20,22-trienolide and 14beta-hydroxybufa-3,5,20,22-tetraenolide were isolated from the dichloromethane extract of the bulbs of Drimia robusta and the methanol extract of the bulbs of Urginea epigea, respectively. The bulbs of Drimia robusta also yielded several known compounds, 6beta-acetoxy-3beta,8beta,12beta,14beta tetrahydroxybufa-4,20,22-trienolide (12beta-hydroxyscillirosidin) from the dichloromethane extract and three common aromatic acids, 4-hydroxy-3 methoxybenzoic acid, 3,4-dihydroxybenzoic acid, and trans-3-(4'-hydroxyphenyl)-2 propenoic acid from the ethyl acetate extract. PMID- 15541734 TI - Limonoids from Cedrela sinensis. AB - Five limonoids were isolated from the leaves of Cedrela sinensis (Meliaceae) and their structures were determined to be 11beta-hydroxy-7alpha-obacunyl acetate, 11 oxo-7alpha-obacunyl acetate, 11-oxo-7alpha-obacunol, 11beta-hydroxycneorin G, and 11-oxocneorin G, by 2D NMR spectroscopic experiments, X-ray crystallographic analysis, and chemical methods. PMID- 15541735 TI - Two labdane diterpenoids and a seco-tetranortriterpenoid from Turreanthus africanus. AB - Examination of the methylene chloride soluble portion of the acetone extract of the seeds of Turreanthus africanus yielded two labdane diterpenoids 12,15 epoxylabda-8(17),12,14-trien-16-al (1) and 16-acetoxy-12(R),15-epoxy-15beta hydroxylabda-8(17),13(16)-diene (2) and a limonoid, 17-epi 12-dehydroxyheudebolin (3). Structures elucidation was based on the analysis of spectroscopic data. PMID- 15541736 TI - Saponins and acylated saponins from Dizygotheca kerchoveana. AB - Four new triterpenoid saponins were isolated from the leaves and stem of branches of Dizygotheca kerchoveana along with seven known ones. The new saponins were respectively characterized as 3-O-[beta-D-glucopyranosyl-(1-->3)]-[beta-D glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl echinocystic acid, 3-O-[beta-D glucopyranosyl-(1-->3)]-[beta-D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl echinocystic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1- >6)-beta-D-glucopyranosyl] ester, 3-O-[beta-D-3-O-trans-p-coumaroyl glucopyranosyl-(1-->3)]-[beta-D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl echinocystic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1- >6)-beta-D-glucopyranosyl] ester and 3-O-[beta-d-3-O-cis-p-coumaroyl glucopyranosyl-(1-->3)]-[beta-D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl echinocystic acid 28-O-[alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1- >6)-beta-D-glucopyranosyl] ester. Their structures were elucidated by 1D and 2D NMR experiments, FAB-MS as well as chemical means. PMID- 15541737 TI - Taxanes with C-5-amino-side chains from the needles of Taxus canadensis. AB - Five taxanes with an amino-side chain on C-5 were identified for the first time in the needles of the Canadian yew, Taxus canadensis. Their structures were characterized as 2alpha,7beta,9alpha,10beta,13-pentaacetoxy-11beta-hydroxy-5alpha (3'-N,N-dimethylamino-3'-phenyl)-propionyloxytaxa-4(20),12-diene (1), 2alpha,9alpha-dihydroxy-10beta,13alpha-diacetoxy-5alpha-(3'-methylamino-3' phenyl)-propionyloxytaxa-4(20),11-diene (2), 2alpha17-dihydroxy 9alpha,10beta,13alpha-triacetoxy-5alpha-(3'-N,N-dimethylamino-3'-phenyl) propionyloxytaxa-4(20),11-diene (3), 2alpha-hydroxy-7beta,9alpha,10beta,13alpha tetraacetoxy-5alpha-(2'-hydroxy-3'-N,N-dimethylamino-3'-phenyl)-propionyloxytaxa 4(20),11-diene (4), and 9alpha-hydroxy-2alpha,10beta,13alpha-triacetoxy-5alpha (3'-N,N-dimethylamino-3'-phenyl)-propionyloxytaxa-4(20),11-diene (5) on the basis of 1D-, 2D-NMR spectroscopic data and high-resolution fast atom bombardment MS analyses. Metabolite (1) was isolated from the needles of the Canadian yew for the first time but had previously been detected in the stems of the Japanese yew, whereas taxanes (2-5) are only now reported. Metabolite (3) is the first reported nitrogen-containing taxane with a 17-hydroxyl substitution. PMID- 15541738 TI - Germacranolides from Calea urticifolia. AB - Four germacranolides, named calealactones A-C and calealactone B [corrected] were isolated from the leaves of Calea urticifolia (Compositae) in addition to three known germacranolides. Their structures were established on the basis of spectroscopic analysis including by 2D NMR spectroscopy. Calealactone C and 2,3 epoxyjuanisulamin displayed potent toxicity to U937 cells. PMID- 15541739 TI - Corsifurans A-C, 2-arylbenzofurans of presumed stilbenoid origin from Corsinia coriandrina (Hepaticae). AB - Chemical investigation of the diethyl ether extract from the liverwort Corsinia coriandrina resulted in the isolation of a new 2-arylbenzofuran compound called corsifuran A. The structure was identified by spectroscopic techniques and confirmed by synthesis. Two minor constituents of similar structure, and two related stilbenoids and a bibenzyl were identified by comparison of the mass spectra and GC retention indices with authentic samples. Due to the similarity in substitution patterns a stilbenoid origin of the corsifurans is proposed. PMID- 15541740 TI - Three isocoumarins and a benzofuran from the cultured lichen mycobionts of Pyrenula sp. AB - The spore-derived mycobionts of the lichen Pyrenula sp. were cultivated on a malt yeast extract medium supplemented with 10% sucrose. The investigation of their metabolites resulted in isolation of four compounds, three isocoumarins and a biogenetically related benzofuran; their structures were determined by spectroscopic methods. PMID- 15541741 TI - Two aurone glycosides from heartwood of Pterocarpus santalinus. AB - Two new aurone glycosides, 6 hydroxy 5 methyl 3',4',5' trimethoxy aurone 4-O alpha-L-rhamnopyranoside and 6,4' dihydroxy aurone 4-O-rutinoside have been isolated from the ethanolic extract of the wood of Pterocarpus santalinus. Their structures were determined on the basis of chemical and spectroscopic analysis (UV, IR, EIMS, (1)H and (13)C NMR). PMID- 15541742 TI - Flavonoids from shoots, roots and roots exudates of Brassica alba. AB - Analysis of extracts obtained from shoots, roots and exudates of Brassica alba revealed the presence of 3,5,6,7,8-pentahydroxy-4'-methoxy flavone in shoots, as well as 2',3',4',5',6'-pentahydroxy chalcone and 3,5,6,7,8-pentahydroxy flavone in roots and exudates. Apigenin was also found in the shoots and roots, but not in the root exudates. PMID- 15541743 TI - Guangsangons F-J, anti-oxidant and anti-inflammatory Diels-Alder type adducts, from Morus macroura Miq. AB - Five Diels-Alder type adducts, named guangsangons F, G, H, I, and J, along with two known compounds, mulberrofuran J and kuwanon J, were isolated from Morus macroura Miq. Their structures were determined by means of spectroscopic analyses and chemical methods. These compounds were regarded biogenetically as Diels-Alder type adducts of dehydroprenylphenols and chalcone derivatives, and (1)H NMR variable temperature experiments suggested that they all existed as an equilibrium mixture of conformational isomers in solution. Among the isolated new compounds, guangsangons H, I, and J displayed potent antioxidant activities and moderate anti-inflammatory activities. PMID- 15541744 TI - Augustamine type alkaloids from Crinum kirkii. AB - Sixteen more Amaryllidaceae alkaloids have been isolated from bulbs of Crinum kirkii Baker of which noraugustamine and 4a,N-dedihydronoraugustamine are hitherto unknown. Their structures and those of earlier known alkaloids have been established by physical and spectroscopic analysis. Application of 2D NMR techniques was used for complete characterization of the alkaloids as well as of 3-O-acetylsanguinine. 1,2-Diacetyllycorine and 3-O-acetylsanguinine showed activity against Trypanosoma brucci rhodesiense, the parasite associated with sleeping sickness. 3-O-acetylsanguinine also showed some activity against Trypanosoma cruzi. PMID- 15541745 TI - In vitro anti-herpetic activity of sulfated polysaccharide fractions from Caulerpa racemosa. AB - A sulfated polysaccharide fraction was isolated from the hot water extract of the green alga Caulerpa racemosa and designated HWE. This polymer, which contained galactose, glucose, arabinose and xylose as the major component sugars, had [alpha](D)(30) + 46.2 degrees in water and contained 9% sulfate hemiester groups. Sugar linkage analysis indicates that HWE was branched and mainly contained 1,3- and 1,3,6-linked galactose, 1,3,4-linked arabinose, 1,4-linked glucose and terminal- and 1,4-linked xylose residues. Sulfation was deduced from infrared spectroscopy and methylation analysis to occur on O-6 of galactose and O-3 of arabinose. The native polysaccharide could be fractionated by size exclusion chromatography into two overlapping fractions and the major fraction has a hydrodynamic volume similar to that of 70 kDa dextran. HWE was a selective inhibitor of reference strains and TK(-) acyclovir-resistant strains of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) in Vero cells, with antiviral effective concentration 50% (EC(50)) values in the range of 2.2-4.2 microg/ml and lacking cytotoxic effects. Furthermore, HWE did not exhibit anticoagulant properties at concentrations near the EC(50). PMID- 15541746 TI - C31-C34 methylated squalenes from a Bolivian strain of Botryococcus braunii. AB - Three new triterpenes, synthesized by a Bolivian strain of the green microalga Botryococcus braunii, were isolated and their chemical structures determined by 1D and 2D NMR, and mass spectrometry. These compounds are tri-, di-, and mono methylsqualenes, co-occurring with the previously identified tetramethylsqualene and some C(30)-C(32) botryococcenes. In this strain, methylated squalenes constitute up to 24% of the total hydrocarbons and 4.5% of the dry biomass. The results of a pulse-chase experiment with L-[Me-(13)C] methionine provide evidence for the origin of these compounds via methylation of squalene at positions 3, 7, 18 and 22. PMID- 15541747 TI - Quassinoid glucosides from seeds of Brucea amarissima. AB - Quassinoid glucosides, javanicosides I, J, K and L, were isolated from the seeds of Brucea amarissima (Lour.) Desv. ex B. A. Gomes (Simaroubaceae), along with two known quassinoids, i.e. bruceins D and E, and seven known quassinoid glucosides, yadanziosides B, C, E, I and K, bruceoside B and yadanzigan. Their structures were elucidated by analysis of the spectral data and chemical evidence. PMID- 15541748 TI - Can mode of action predict mixture toxicity for risk assessment? AB - Recent regulatory guidance for mixture risk assessments and for regulating pesticide chemicals recommends using information about the "mode" or "mechanism" of action of individual chemicals to predict dose response characteristics of mixtures. Dose addition is assumed for mixtures of chemicals that have similar mechanisms and response addition for those with dissimilar mechanisms. Three different sets of criteria have been formulated to guide the selection of an appropriate data set for characterizing a chemical's mode of action, but the sufficiency of those criteria to predict dose addition for a mixture has not been validated experimentally. Several examples from the pharmacological and toxicological literature challenge the premise that dose response characteristics of a mixture can be predicted from the modes of action of its components. Detoxification pathways may need to be understood before dose addition in the observable effect range can be extrapolated to mixture concentrations below the no observable effect levels of the mixture components. Because elucidating discreet mechanisms of action may be possible only for chemicals that exhibit a high degree of biological specificity and dose sensitivity, practical limitations on the approach must be defined. To reduce the large uncertainties inherent in the recommended approach, future research should be focused on defining the mechanistic features that predict dose additive toxicity in mixtures. A detailed characterization of pharmacodynamics, pharmacokinetics, and slope of dose response curves may be necessary to evaluate whether the toxicity of a mixture can be predicted by the mode of action of its component chemicals. PMID- 15541749 TI - Involvement of glial cells in the neurotoxicity of parathion and chlorpyrifos. AB - An in vitro model, the aggregating brain cell culture of fetal rat telencephalon, has been used to investigate the influence of glial cells on the neurotoxicity of two organophosphorus pesticides (OPs), chlorpyrifos and parathion. Mixed-cell aggregate cultures were treated continuously for 10 days between DIV 5 and 15. Parathion induced astrogliosis at concentration at which MAP-2 immunostaining, found here to be more sensitive than neuron-specific enzyme activities, was not affected. In contrast, chlorpyrifos induced a comparatively weak gliotic reaction, and only at concentrations at which neurons were already affected. After similar treatments, increased neurotoxicity of parathion and chlorpyrifos was found in aggregate cultures deprived of glial cells. These results suggest that glial cells provide neuroprotection against OPs toxicity. To address the question of the difference in toxicity between parathion and chlorpyrifos, the toxic effects of their leaving groups, p-nitrophenol and trichloropyridinol, were studied in mixed-cell aggregates. General cytotoxicity was more pronounced for trichloropyridinol and both compounds had similar toxic effects on neuron specific enzyme activities. In contrast, trichloropyridinol induced a much stronger decrease in glutamine synthetase activity, the enzymatic marker of astrocytes. Trichloropyridinol may exert a toxic effect on astrocytes, compromising their neuroprotective function, thus exacerbating the neurotoxicity of chlorpyrifos. This is in line with the suggestion that glial cells may contribute to OPs neurotoxicity, and with the view that OPs may exert their neurotoxic effects through different mechanisms. PMID- 15541750 TI - Effect of mitoguazone on polyamine oxidase activity in rat liver. AB - Mitoguazone is a known inhibitor of polyamine biosynthesis through competitive inhibition of S-adenosylmethionine decarboxylase. A recent renewed interest in mitoguazone as an antineoplastic agent prompted us to investigate the effect of the drug on polyamine catabolism in rat liver, since the organ plays an important role in detoxification mechanisms. Thus, the purpose of this work was to evaluate the effect of in vivo mitoguazone administration on polyamine catabolic enzymes. In particular, our interest was directed to the changes in polyamine oxidase activity, since this enzyme has been recently confirmed to exert important functions that until now were underestimated. Mitoguazone administration induced hepatic polyamine oxidase activity starting at 4 h after administration, and the enzyme returned to basal levels 96 h after treatment. The changes in enzyme activity were accompanied by changes in putrescine concentrations, which increased starting at 4 h until 72 h after treatment. We also evaluated the activity of the newly identified spermine oxidase, which was not significantly changed by mitoguazone treatment. Therefore, we hypothesized that the enzyme involved in mitoguazone response of the liver is the polyamine oxidase, which acts on acetylated polyamines as substrate. PMID- 15541751 TI - CYP3A in horse intestines. AB - The intestinal enterocytes provide the initial site for cytochrome P450 (CYP) mediated metabolism of orally absorbed xenobiotics. In man and some animal species, the CYP3A subfamily is highly expressed in the intestines and considered to be important in the first-pass metabolism of drugs and other xenobiotics. The aim of the present study was to investigate the mRNA expression, immunohistochemical localization and catalytic activity of CYP3A in the intestines of horse. Real-time RT-PCR analyses showed that the highest CYP3A mRNA expression was present in the duodenum with a decreasing level towards jejunum, ileum, cecum, and colon. The CYP3A mRNA expression in the liver was similar as in the anterior part of the jejunum, but about 4.5 times lower than in the anterior part of the duodenum. Immunohistochemistry showed CYP3A immunoreactivity in the cytoplasm of the enterocytes, which decreased distally along the intestinal tract. CYP3A-dependent metabolic activity rose slightly from the anterior to the distal part of the duodenum and the anterior part of the jejunum and then declined to the middle and distal parts of the jejunum and the ileum, cecum, and colon. Our results suggest that CYP3A in the small intestine plays a major role in first-pass metabolism and may affect bioavailability and therapeutic efficiency of some orally administrated drugs in horse. PMID- 15541752 TI - Proteomic analysis of acrylamide-protein adduct formation in rat brain synaptosomes. AB - Evidence suggests that the neurological defects (gait abnormalities, foot splay, and skeletal muscle weakness) associated with acrylamide (ACR) intoxication are mediated by impaired neurotransmission at central and peripheral synapses. ACR can form adducts with nucleophilic residues on proteins and thereby alter corresponding structure and function. To evaluate protein adduction in nerve terminals as a possible mechanism of action, recombinant N-ethylmaleimide sensitive factor (NSF) was exposed in vitro to ACR (10 micromol) and mass spectrometry (MS) was used to identify adduct sites. MS analyses demonstrated that ACR formed adducts with sulfhydryl groups on cysteine residues (carbamoylethylcysteine, or CEC) of NSF. Ex vivo incubation of whole brain synaptosomes with ACR (0.001-1.0 M) produced concentration-dependent increases in CEC that were inversely correlated to reductions in neurotransmitter release that occurred over the same neurotoxicant concentration range. In synaptosomes isolated from rats intoxicated at a higher (50 mg/kg per day x 3, 5, 8, or 11 days) or a lower (21 mg/kg per day x 14, 21, or 28 day) ACR dose rate, CEC levels increased progressively up to a moderate level of neurotoxicity. To identify protein adducts, synaptosomal proteins labeled by ex vivo 14C-ACR exposure were separated by gel electrophoresis and probed by immunoblot analysis. Results showed that NSF and the SNARE protein, SNAP-25, were tentative ACR targets. Subsequent experiments indicated that ACR exposure increased synaptosomal levels of the 7S SNARE core complex, which is consistent with inhibition of NSF, SNAP-25 function, or both. These data suggest that adduction of cysteine residues on NSF and certain SNARE proteins might be causally involved in the nerve terminal dysfunction induced by ACR. PMID- 15541753 TI - Target-specific action of organochlorine compounds in reproductive and nonreproductive tissues of estrogen-reporter male mice. AB - Organochlorines are lipophylic molecules that accumulate in the fat where they remain for years. During weight loss, they are mobilized and their concentration increases in blood. The present work tests, in transgenic estrogen-reporter mice (ERE-tK-LUC), whether this increase is sufficient to modulate the estrogen receptors (ERs) in the whole body. Three weak estrogens were studied: p,p'DDT [1,1,1-trichloro2,2-bis(p-chlorophenyl) ethane], p,p'DDE [1,1-dichloro-2,2-bis(p chlorophenyl)ethylene], and betaBHC [beta-benzene-hexachloride]. Dose-dependent analysis of reporter expression (luciferase) were performed in tissues of acutely treated mice. A body map of ER activation was obtained. All these chemicals modulated the reporter, although with a different efficiency and depending upon the tissue analyzed. Induction was confirmed in the liver by determining the expression of the endogenous progesterone receptor (PR) gene, at the dose and time point at which the luciferase gene was maximally induced. After experimental accumulation in the fat tissue, followed by a 48-h period of fasting, we tested whether these compounds could be mobilized to reach sufficient levels to activate the ERs in selected reproductive and nonreproductive tissues (testicle, prostate, liver, and lung). This experimental setting produced results that were different than those obtained following acute treatments. In loaded mice, fasting induced betaBHC mobilization resulted in strong ER activation in the liver and the lung, which was blocked by ICI-182780. p,p'DDT mobilization had no effect in these tissues, but it acted efficiently in the prostate and testis. betaBHC inhibited the ERE-mediated reporter in the testicle and induced the reporter in the prostate. In this tissue, betaBHC action was not inhibited by the anti-estrogen ICI-182780. During fasting, betaBHC, p,p'DDT, and metabolite p,p'DDE increased in blood concentration, from 2.25 +/- 0.25, 0.51 +/- 0.09, and 0.38 +/- 0.06 microg/ml to 8.24 +/- 0.95, 4.52 +/- 0.68, and 5.06 +/- 0.57 microg/ml, respectively. The effect produced by these organochlorines in the liver correlates with the modulation of the ERalpha protein. We conclude that these organochlorines modulate differently the expression of estrogen-regulated genes in male mice. Their effect is tissue- and compound-specific and is dependent on the energetic balance. PMID- 15541754 TI - Zinc might protect oxidative changes in the retina and pancreas at the early stage of diabetic rats. AB - It is well documented that oxidative stress is a basic mechanism behind the development of diabetic retinopathy (DR). The current study was undertaken to elucidate the possible role of zinc as an antioxidant and a biological membrane stabilizer in the protection against (DR). Male Wistar rats weighing 250 +/- 50 g were made diabetic by injection with a single ip dose of alloxan (100 mg/kg). Another group of rats was simultaneously treated with alloxan (100 mg/kg) and a single ip dose of zinc chloride (ZnCl2) (5 mg/kg). Blood and tissue samples were collected at 24, 48, and 72 h post-treatment in both groups. Diabetic state was confirmed by the determination of plasma glucose levels (significantly elevated at any time of the experiment when compared with controls receiving vehicle). Plasma insulin was significantly increased 24 h after treatment in both alloxan and alloxan plus ZnCl2-treated groups, and then decreased markedly 48 and 72 h post treatment in both groups. Alloxan treatment depleted both retinal and liver glutathione contents. The decrease in retinal and liver GSH in alloxan-treated rats was accompanied with a sustained increase in their thiobarbituric acid (TBA) content. Simultaneous treatment of rats with alloxan and ZnCl2 blunted the sustained increment in plasma glucose induced by alloxan. The combined administration of alloxan and zinc reversed the depleting effect on retinal and hepatic GSH in alloxan-treated rats and reduced the elevations in TBA content of both retinas and livers. At variance with many other antioxidants the current results clearly indicate the beneficial effects of Zn in both controlling hyperglycemia and the protection of the retina against oxidative stress in diabetes which may help set a new direction toward the development of effective treatments of DR. PMID- 15541755 TI - Uptake of inorganic and organic derivatives of arsenic associated with induced cytotoxic and genotoxic effects in Chinese hamster ovary (CHO) cells. AB - Humans are exposed to arsenic and their organic derivatives, which are widely distributed in the environment, via food, water, and to a lesser extent, via air. Following uptake, inorganic arsenic undergoes biotransformation to mono- and dimethylated metabolites. Recent findings suggest that the methylation reactions represent a toxification rather than a detoxification pathway. In the present study, the genotoxic effects and the cellular uptake of inorganic arsenic [arsenate, As(i)(V); arsenite, As(i)(III)] and the methylated arsenic species monomethylarsonic acid [MMA(V)], monomethylarsonous acid [MMA(III)], dimethylarsinic acid [DMA(V)], dimethylarsinous acid [DMA(III)], trimethylarsenic oxide [TMAO(V)] were investigated in Chinese hamster ovary (CHO-9) cells. The chemicals were applied at different concentrations (0.1 microM to 10 mM) for 30 min and 1 h, respectively. Cytotoxic effects were investigated by the trypan blue extrusion test and genotoxic effects by the assessment of micronucleus (MN) induction, chromosome aberrations (CA), and sister chromatid exchanges (SCE). Intracellular arsenic concentrations were determined by ICP-MS techniques. Our results show that MMA(III) and DMA(III) induce cytotoxic and genotoxic effects to a greater extent than MMA(V) or DMA(V). Viability was significantly decreased after incubation (1 h) of the cells with > or = 1 microM As(i)(III), > or = 1 microM As(i)(V), > or = 500 microM MMA(III), > or = 100 microM MMA(V), and 500 microM DMA(V) and > or = 0.1 microM DMA(III). TMAO(V) was not cytotoxic at concentrations up to 10 mM. A significant increase of the number of MN, CA and SCE was found for DMA(III) and MMA(III). As(i)(III + V) induced CA and SCE but no MN. TMAO(V), MMA(V) and DMA(V) were not genotoxic in the concentration range tested (up to 5 mM). The nuclear division index (NDI) was not affected by any of the tested arsenic compounds after a recovery period of 14 to 35 h. When the uptake of the chemicals was measured by ICP-MS analysis, it was found that only 0.03% MMA(V) and DMA(V), and 2% MMA(III), As(i)(III) and (V) were taken up by the cells. In comparison, 10% of the DMA(III) dose was taken up. The total intracellular concentration of all arsenic compounds increased with increasing arsenic concentrations in the culture medium. Taken together, these data demonstrate that arsenic compounds in the trivalent oxidation state exhibit the strongest genotoxic effects. Trivalent organoarsenic compounds are more membrane permeable than the pentavalent species. The potency of the DNA damage decreases in the order DMA(III) > MMA(III) > As(i)(III and V) > MMA(V) > DMA(V) > TMAO(V). We postulate that the induction of genotoxic effects caused by the methylated arsenic species is primarily dependent upon their ability to penetrate cell membranes. PMID- 15541756 TI - Interindividual variation in the metabolism of arsenic in cultured primary human hepatocytes. AB - Liver is a prime site for conversion of inorganic arsenic (iAs) to methylated metabolites, including methylarsenicals (MAs) and dimethylarsenicals (DMAs). To assess interindividual variation in the capacity of liver to metabolize iAs, we examined the metabolic fate of arsenite (iAs(III)) in normal primary human hepatocytes obtained from eight donors and cultured under standard conditions. Methylation rates, yields, and distribution of arsenicals were determined for hepatocytes exposed to 0.3-30 nmol of iAs(III)/mg of protein for 24 h. Although the accumulation of arsenic (As) by cells was a linear function of the initial concentration of iAs(III) in culture, the concentration of As retained in cells varied several fold among donors. DMAs was the major methylated metabolite found in cultures exposed to low concentrations of iAs(III); at higher concentrations, MAs was always predominant. Maximal rates for methylation of iAs(III) were usually attained at 3 or 9 nmol of iAs(III)/mg of protein and varied about 7-fold among donors. For most donors, the methylation rate decreased at the highest iAs(III) concentrations. MAs was the major methylated metabolite retained in cells regardless of exposure level. DMAs was the major methylated metabolite found in medium. The interindividual differences in rates for iAs(III) methylation were not strictly associated with variations in basal mRNA levels for cyt19, an As-methyltransferase. Analysis of the coding sequence of cyt19 identified one heterozygote with Met287Thr mutation in a single allele. Thus, genetic polymorphism of cyt19 along with other cellular factors is likely responsible for interindividual differences in the capacity of primary human hepatocytes to retain and metabolize iAs(III). PMID- 15541757 TI - cDNA microarray analysis of rat alveolar epithelial cells following exposure to organic extract of diesel exhaust particles. AB - Diesel exhaust particles (DEP) induce pulmonary diseases including asthma and chronic bronchitis. Comprehensive evaluation is required to know the mechanisms underlying the effects of air pollutants including DEP on lung diseases. Using a cDNA microarray, we examined changes in gene expression in SV40T2 cells, a rat alveolar type II epithelial cell line, following exposure to an organic extract of DEP. We identified candidate sensitive genes that were up- or down-regulated in response to DEP. The cDNA microarray analysis revealed that a 6-h exposure to the DEP extract (30 microg/ml) increased (>2-fold) the expression of 51 genes associated with drug metabolism, antioxidation, cell cycle/proliferation/apoptosis, coagulation/fibrinolysis, and expressed sequence tags (ESTs), and decreased (<0.5-fold) that of 20 genes. In the present study, heme oxygenase (HO)-1, an antioxidative enzyme, showed the maximum increase in gene expression; and type II transglutaminase (TGM-2), a regulator of coagulation, showed the most prominent decrease among the genes. We confirmed the change in the HO-1 protein level by Western blot analysis and that in the enzyme activity of TGM-2. The organic extract of DEP increased the expression of HO-1 protein and decreased the enzyme activity of TGM-2. Furthermore, these effects of DEP on either HO-1 or TGM-2 were reduced by N-acetyl-l-cysteine (NAC), thus suggesting that oxidative stress caused by this organic fraction of DEP may have induced these cellular responses. Therefore, an increase in HO-1 and a decrease in TGM-2 might be good markers of the biological response to organic compounds of airborne particulate substances. PMID- 15541758 TI - Antitumor effects of the partially purified polysaccharides from Antrodia camphorata and the mechanism of its action. AB - Antrodia camphorata is a popular folk medicine that has attracted great attention due to its fame for antitumor activity against cancer. However, there is little information available about its action. In the present study, we purified a unique polysaccharide component from A. camphorata mycelia (AC-PS) and found that it has pronounced anti-tumor effects on both in vitro and in vivo model. Our results showed that AC-PS alone did not show any direct cytotoxic effect to human leukemic U937 cells, even at high concentration (200 microg/ml). However, it could inhibit the proliferation of U937 cells via activation of mononuclear cells (MNCs). Treatment of U937 cells with AC-PS-stimulated-MNC-CM could significantly inhibit its proliferation with 55.3% growth inhibition rate. The in vitro antitumor activity was substantiated by the in vivo therapeutical study of AC-PS in sarcoma 180-bearing mice. Intraperitoneal and oral administration of AC-PS, 100 and 200 mg/kg significantly suppressed the tumor growth with the inhibition rate of 69.1% and 58.8%, respectively. In vivo studies also showed that several immunoparameters, such as the spontaneous proliferation of spleen cells, after AC PS administration, were two-fold higher than in control mice. Furthermore, the cytolytic activity of spleen cells also increased from 9.8 +/- 1.1% in control mice to 34.2 +/- 5.5% and 48.2 +/- 2.5%, after oral and intraperitoneal treatment, respectively. Besides, the mice serum interleukin-12 levels increased significantly by AC-PS treatment. Considering all these results, it is suggested that AC-PS elicit its anti-tumor effect by promoting a Th1-dominant state and killer activities. PMID- 15541759 TI - The effects of benzene and the metabolites phenol and catechol on c-Myb and Pim-1 signaling in HD3 cells. AB - Exposure to the environmental toxicant benzene has been proposed to lead to leukemogenesis. The transcription factor c-Myb plays a role in blood cell differentiation and can be regulated by the serine-threonine kinase Pim-1. Overexpressed versions of c-Myb and Pim-1 are believed to play a key role in the development of a wide variety of leukemias and tumors. In our study, we evaluated the effects of benzene and the metabolites catechol and phenol on c-Myb signaling to investigate our hypothesis that benzene exerts its toxicity by interfering with this pathway. To evaluate this hypothesis, HD3 chicken erythroblast cells were transiently transfected with a c-Myb responsive luciferase reporter plasmid and then exposed to benzene, catechol, or phenol (0-300 microM) for 1-24 h before nonproprietary dual luciferase activities were measured. Our results demonstrated that catechol exposure caused a time- and concentration-dependent increase in c Myb activity with significance occurring at 100 and 300 microM after 24 h of exposure, which was independent of increased Pim-1 protein, but dependent on increased c-Myb phosphorylation. Benzene and phenol exposure resulted in small but significant decreases in c-Myb activity that were not dose- and time dependent, nor was increased Pim-1 protein involved. These results are consistent with other studies, which suggest metabolite differences in benzene-mediated toxicity. More importantly, this study supports the hypothesis that benzene may mediate its toxicity through metabolite-mediated alterations in the c-Myb signaling pathway. PMID- 15541762 TI - A contemporary understanding of progesterone receptor function. AB - Following the discovery of coactivators for nuclear receptors and the identification of a significant number of progesterone receptor (PR) target genes, our understanding of PR action has extended from its normally recognized functions to a wide variety of seemingly unlinked biological processes. Recent advances in defining the action of PR coactivators has revealed important mechanisms involving multiple layers of regulation in PR-mediated transcription. In this review, we will discuss the current understanding of PR physiology and the molecular basis of coactivator function in PR signaling events. PMID- 15541763 TI - A two-site model for antiestrogen action. AB - Evidence is presented for a unified model for the reaction of antiestrogens with estrogen receptors that explains much of the unusual pharmacology of these clinically important agents. Agonist activity results from occupancy of the estradiol-binding (primary) site in the receptor and antagonism from the additional interaction with a secondary locus not recognized by hormone. In the case of type I antiestrogens, such as tamoxifen, this is weaker than primary site binding, so these substances are agonists at low concentrations and antagonists at higher levels. With type II antiestrogens, affinities for both sites are comparable, so one never has the agonist situation (only primary site occupied), and these agents are pure antagonists. Reproductive tissues of the mouse and guinea pig contain a small macromolecule that binds hydroxytamoxifen, but not estradiol, keeping the free antiestrogen concentration below that required for secondary binding. Thus, in these species, tamoxifen is a pure agonist. PMID- 15541764 TI - Partial androgen insensitivity with phenotypic variation caused by androgen receptor mutations that disrupt activation function 2 and the NH(2)- and carboxyl terminal interaction. AB - Partial androgen insensitivity with sex phenotype variation in two unrelated families was associated with missense mutations in the androgen receptor (AR) gene that disrupted the AR NH(2)-terminal/carboxy terminal interaction. Each mutation caused a single amino acid change within the region of the ligand binding domain that forms activation function 2 (AF2). In one family, the mutation I737T was in alpha helix 4 and in the other F725L was between helices 3 and 4. Neither mutation altered androgen binding as determined by assays of mutant AR in the patient's cultured genital skin fibroblasts or of recombinant mutant receptors transfected into COS cells. In transient cotransfection assays in CV1 cells, transactivation with the AR mutants at low concentrations of DHT was reduced several fold compared with wild-type AR but increased at higher concentrations. Defects in NH(2)-terminal/carboxy terminal interactions were identified in mammalian two hybrid assays. In similar assays, there was reduced binding of the p160 coactivators TIF2/SRC2 and SRC1 to the mutant AR ligand binding domains (LBD). In the family with AR I737T, sex phenotype varied from severely defective masculinization in the proband to a maternal great uncle whose only manifestation of AIS was severe gynecomastia. He was fertile and passed the mutation to two daughters. The proband of the F725L family was also incompletely masculinized but was raised as a male while his half-sibling by a different father was affected more severely and reared as a female. These studies indicate that the function of an AR AF2 mutant in male development can vary greatly depending on the genetic background. PMID- 15541765 TI - Mechanisms of glucocorticoid receptor signaling during inflammation. AB - Glucocorticoids are among the most widely prescribed anti-inflammatory drugs. They act by binding to the glucocorticoid receptor (GR) that, upon activation, translocates to the nucleus and either stimulates or inhibits gene expression. GR inhibition of many proinflammatory response genes occurs through induction of the synthesis of anti-inflammatory proteins as well as through repression of proinflammatory transcription factors, such as nuclear factor-kappaB (NF-kappaB) or activator protein-1 (AP-1). In this review, we discuss the molecular mechanisms underlying GR inhibition of inflammatory responses, with an emphasis on repression of NF-kappaB and AP-1 and their respective signaling pathways. PMID- 15541766 TI - Convergence of lipid homeostasis through liver X and thyroid hormone receptors. AB - Members of the nuclear receptor gene family act as biological rheostats to maintain metabolic homeostasis in response to endocrine and nutritional changes. The liver X (LXR) and thyroid hormone (TR) receptors have been shown to regulate overlapping but distinct metabolic pathways important for overall lipid homeostasis. Dyslipidemia is one out of four key determinants for cardiovascular risk and both LXRs and TRs may provide attractive targets for intervention of cardiovascular disease. In this review we will compare the two receptor systems to highlight similarities and differences in structure and function with implications for development of novel treatments for dyslipidemia and atherosclerosis. PMID- 15541767 TI - The regulation of COUP-TFII gene expression by Ets-1 is enhanced by the steroid receptor co-activators. AB - Recent phenotypic analysis of orphan nuclear receptor chicken ovalbumin upstream promoter-transcription factor II (COUP-TFII) [NR2F2] knockout mice shows that COUP-TFII is involved in the angiogenic process in the developing embryos. Since Ets-1 expression is also correlated with angiogenesis, and both Ets-1 and COUP TFII mRNA are present in mesenchymal cells, we have sought to determine whether Ets-1 is a potential regulator of COUP-TFII gene expression. For this purpose, we performed transient transfection experiments using a luciferase reporter construct containing the mouse COUP-TFII promoter. We found that the COUP-TFII promoter activity is indeed regulated by Ets-1. We have identified two identical inverted potential ETS-binding sites located 47 nucleotides downstream of the start site. Mutation of both sites reduced the ability of Ets-1 to enhance the COUP-TFII promoter activity. Furthermore, other members of the ETS family such as Ets-2 or ETV1 are also potent regulators of the COUP-TFII promoter. Finally, the induction of the COUP-TFII gene is strongly enhanced by the expression of steroid receptor co-activator factors through a direct interaction with Ets-1. These results indicate that COUP-TFII is a potential downstream target of Ets-1 and it may partially mediate the Ets-1 function in angiogenesis. PMID- 15541768 TI - Gene regulation for the senescence marker protein DHEA-sulfotransferase by the xenobiotic-activated nuclear pregnane X receptor (PXR). AB - Dehydroepiandrosterone (DHEA)-sulfotransferase (SULT2A1) is a phase II metabolizing/detoxifying enzyme with substrate preference for physiological hydroxysteroids, diverse drugs and other xenobiotics. The first-pass tissues (liver and intestine) express SULT2A1 at high levels. In senescent male rodents, Sult2A1 gene transcription in the liver is markedly enhanced and calorie restriction retards this increase. Age-associated loss of the liver expression of androgen receptor in part explains the up-regulation of Sult2A1 expression at late life, since androgen receptor is a negative regulator of this gene. In line with its role in xenobiotic metabolism, the Sult2A1 gene is induced by the pregnane X receptor (PXR). PXR is a xenosensing nuclear receptor that is activated by endobiotic (natural steroids) and xenobiotic (therapeutic drugs and environmental chemicals) molecules. An inverted-repeat arrangement (IR0) of the consensus half site binding sequence for nuclear receptors mediates the xenobiotic induction of the Sult2A1 promoter. The IR0 element is a specific binding site for PXR and its heterodimer partner retinoid X receptor (RXR-alpha) and it directs PXR-mediated induction of a heterologous promoter. In contrast to the loss of androgen receptor expression, PXR and RXR-alpha mRNA expression is invariant during aging. Repression by the androgen receptor and induction by PXR may act coordinately to cause the senescence associated and xenobiotic mediated stimulation of Sult2A1 transcription. Increased Sult2A1 expression appears to be an adaptive response to ensure optimal metabolism of Sult2A1 substrates at old age. PMID- 15541769 TI - Impact of genome instability on transcription regulation of aging and senescence. AB - Genomic instability has been implicated as a major stochastic mechanism of aging. Using a transgenic mouse model with chromosomally integrated lacZ mutational target genes, mutations were found to accumulate with age at an organ- and tissue specific rate. Also, the spectrum of age-accumulated mutations was found to differ greatly from organ-to-organ; while initially similar, mutation spectra of different tissues diverged significantly over the lifetime. To explain how genomic instability, which is inherently stochastic, can be a causal factor in aging, it is proposed that randomly induced mutations may adversely affect normal patterns of gene regulation, resulting in a mosaic of cells at various stages on a trajectory of functional decline, eventually resulting in cell death or neoplastic transformation. To directly address this question, we demonstrate that it is now possible to analyze single cells, isolated from old and young tissues, for specific alterations in gene expression. PMID- 15541770 TI - Age-dependent modulation of DNA repair enzymes by covalent modification and subcellular distribution. AB - Chronic oxidative stress is generally believed to be a major etiologic factor in the aging process. In addition to modulation of signaling processes and oxidation of cellular proteins and lipids, reactive oxygen species (ROS) induce multiple damages in both nuclear and mitochondrial genomes, most of which are repaired via the DNA base excision repair pathway. 8-Oxoguanine (8-oxoG), a major ROS product in the genome, is excised by 8-oxoG-DNA glycosylase (OGG1) and the resulting abasic (AP) site is cleaved by AP-endonuclease (APE1) in the initial steps of repair. Here, we provide data showing that differences between young and aged cells' efficiency in import of OGG1 and APE1 may be responsible for age associated increase in DNA damage in both nuclear and mitochondrial compartments. It is also evident that age-dependent changes in covalent modifications of APE1 by acetylation regulate its action as a transcriptional repressor of many Ca(2+) responsive genes by binding to nCaRE, in addition to its endonuclease activity. Thus, ROS-induced altered signaling is responsible for age-dependent changes in post-translational modifications and import of DNA repair enzymes into nuclei and mitochondria (mt), which in turn affect repair of their genomes. PMID- 15541771 TI - Novel regulatory factors of HSF-1 activation: facts and perspectives regarding their involvement in the age-associated attenuation of the heat shock response. AB - An attenuated response to stress is characteristic of senescence. Heat shock (HS), a significant form of stress, is delayed and reduced in aging organisms. In the response to heat shock, heat shock factor 1 (HSF-1) is activated by trimerization of its monomeric subunits. This then initiates the transcription of a series of heat shock genes (hsp genes) that encode chaperone proteins protective against heat stress. Using a promoter binding electromobility shift assay (EMSA), we have found no activation of this transcription factor in the brains of old (36 months) rats in response to exposure to 41 degrees C for 1h while strong activation is elicited in young (6 months) animals. Since brains of young and old rats had approximately the same amount of HSF-1 subunits, we anticipated the presence of auxiliary regulatory factors essential for the activation of HSF-1 and the initiation of heat shock gene transcription. We describe three novel auxiliary factors--the proteins I-HSF [HSF inhibitor] and elongation factor-1 alpha (EF-1alpha) and a large non-coding RNA (HSR)--that participate in regulation and activation of HSF-1 in early stages of heat shock gene transcription. I-HSF inhibits trimerization of HSF-1 at normal temperatures. HSR and EF-1alpha form a complex with HSF-1 and facilitate its trimerization and binding to heat shock element (HSE) in the promoters of hsps. It is proposed that structural changes in any one or a combination of these factors in response to heat shock may contribute to the age-associated attenuation in the response to stress. PMID- 15541772 TI - Senescent fibroblasts resist apoptosis by downregulating caspase-3. AB - In replicative senescence, cells undergo permanent exit from cell cycle traverse; this is traditionally thought to occur at the end of a culture's in vitro life span, after serial passaging. In general, the checkpoint for replicative senescence is found at the G(1)/S border, controlled by the modulation of a battery of proteins, typified by gaining inhibitors of cell cycle traverse, such as cyclin-dependent kinases or RB hyperphosphorylation, and losing pro proliferation gene expressions such as c-fos, c-myc, and a cadre of proliferation dependent kinases. Here, we present evidence that replicatively senescent fibroblasts are resistant to apoptotic death, associated with a lack of key enzyme activities, caspase-3 being the chief executioner. This observation, coupled with our earlier report that senescent fibroblasts maintain persistently high levels of pro-survival factor Bcl-2, suggests that the molecular signaling program present in fibroblasts at the end of their in vitro life span may not only cater to the state of permanent exit from cell cycle traverse, but also dictate an inability to commit cellular suicide. Future experiments will reveal whether replicatively senescent fibroblasts that can neither proliferate nor die contribute to organismic aging, and whether their accumulation over time in tissue becomes detrimental to the normal aging process. PMID- 15541773 TI - Akt/PKB and p38 MAPK signaling, translational initiation and longevity in Snell dwarf mouse livers. AB - The insulin/IGF-1/GH and p38 MAPK signaling pathways play a key role in the regulation of protein synthesis. The regulation of GH and TSH secretion hormones, that affect the activity of these pathways, plays an important role in the decline of rates of protein synthesis in aged rodent tissues. Studies have indicated that longevity of the Snell dwarf (Pit-1) mouse mutant is associated with the reduction of function of the insulin/IGF-1/GH signaling pathway. We have previously shown that PI3K activity, a signaling protein that plays a key role in the regulation of translation, is also dramatically decreased in the Snell dwarf liver suggesting that the protein synthesis-signaling pathway may be attenuated in this long-lived mouse. Similarly, signaling via p38 MAPK also plays a role in the regulation of protein synthesis. In this study we examined the activities of these signaling pathways to determine if the translation-signaling pathway is altered in young versus aged Snell dwarf mouse livers. Our data indicate that the phosphorylation and kinase activities of Akt/PKB and p38 MAPK, and the levels of phosphorylation of downstream regulators of translation are decreased in dwarf mouse livers. Thus, the overall activities of major components of the translational initiation pathway are decreased in the long-lived Snell dwarf mouse livers. We propose that down-regulation of protein synthesis may be an important characteristic of the Pit-1 mutation and longevity of the Snell dwarf mouse. PMID- 15541774 TI - Nuclear factor kappa B activation by NADPH oxidases. AB - Reactive oxygen species (ROS) initiate activation of the transcription factor NF kappaB in a variety of cell systems. Perhaps the most potent biological source of ROS is the NADPH oxidase of phagocytic cells, a multi-component system that catalyzes the formation of superoxide anion. Although phagocytes use this oxidase to kill ingested microorganisms, the products also mediate a broad range of biological oxidation reactions and some evidence exists for activation of NF kappaB through this mechanism. Moreover, the components of the phagocyte NADPH oxidase are present in certain non-phagocytic cells and recently discovered homologues of the catalytic component gp91(phox) are expressed in a number of tissues. We explored the hypothesis that the products of NADPH oxidases cause the activation of NF-kappaB. K562 human erythrokeukemia cells transfected with constructs for expression of gp91(phox), plus other essential NADPH oxidase components generated substantial amounts of superoxide when activated with phorbol ester, lesser amounts with arachidonic acid exposure, and none with TNFalpha. Gel shift assays demonstrated induction of NF-kappaB in K562 cells exposed to TNFalpha and specificity was shown by oligonucleotide competition. Supershift assays demonstrated the presence in nuclear complexes of the NF-kappaB components p65/RelA and p50. Nuclear complexes of identical electrophoretic mobility were induced in phorbol ester-stimulated K562 cells that expressed the complete NADPH oxidase system, but not in cells lacking one of the essential oxidase components. K562 cells were relatively resistant to NF-kappaB induction by exogenous peroxide, but certain other cell types (HEK293 and HeLaS3) demonstrated such induction upon exposure to reagent hydrogen peroxide or glucose oxidase plus glucose and this was blocked by catalase. Finally, we found a biphasic pattern of gp91(phox) expression in rat liver during aging. High levels observed in young animals decreased in middle age, but increased again in old age. Collectively, these studies demonstrate the potential for NADPH-dependent induction of NF-kappaB and raise the possibility of a role for this pathway in the biology of aging. PMID- 15541775 TI - The role of oxidative damage and stress in aging. AB - The Free Radical/Oxidative Stress Theory of Aging, which was first proposed in 1956, is currently one of the most popular explanations for how aging occurs at the biochemical/molecular level. However, most of the evidence in support of this theory is correlative, e.g., oxidative damage to various biomolecules increases with age, and caloric restriction, which increases life span and retards aging, reduces the age-related increase in oxidative damage to biomolecules. The most direct test of the Free Radical/Oxidative Stress Theory of Aging is to specifically alter the age-related increase in oxidative damage and determine how this alteration affects life span. For the first time, investigators can use genetically altered animals to test directly the role of oxidative damage in aging. In this manuscript, we critically review the past research in this area and discuss potential future research directions in testing the Free Radical/Oxidative Theory of Aging. PMID- 15541776 TI - Replicative senescence: a critical review. AB - Human cells in culture have a limited proliferative capacity. After a period of vigorous proliferation, the rate of cell division declines and a number of changes occur in the cells including increases in size, in secondary lysosomes and residual bodies, nuclear changes and a number of changes in gene expression which provide biomarkers for senescence. Although human cells in culture have been used for over 40 years as models for understanding the cellular basis of aging, the relationship of replicative senescence to aging of the organism is still not clear. In this review, we discuss replicative senescence in the light of current information on signal transduction and mitogenesis, cell stress, apoptosis, telomere changes and finally we discuss replicative senescence as a model of aging in vivo. PMID- 15541777 TI - Cocaine craving and attentional bias in cocaine-dependent schizophrenic patients. AB - Cocaine craving has been implicated as a major factor underlying addiction and drug relapse. From a cognitive viewpoint, craving may reflect, in part, attentional processing biased in favor of drug-related cues and stimuli. Schizophrenic individuals (SZ), however, abuse cocaine in high numbers but typically manifest baseline cognitive deficits that impair their ability to selectively allocate their attentional resources. In this study, we examined the relationship between attentional bias and craving in patients with cocaine dependence (COC; n=20), schizophrenic patients comorbid for cocaine dependence (COC+SZ; n=23), as well as two other comparison groups using a modified version of the Stroop test to include cocaine-relevant words. Results revealed that only the COC patients demonstrated Stroop interference on the cocaine-related words. Moreover, COC patients' attentional processing biases were significantly associated with their cocaine craving severity ratings. COC+SZ patients, in contrast, did not demonstrate Stroop interference and manifested significantly fewer craving symptoms than their COC counterparts. These results suggest that COC+SZ patients' inability to selectively encode their drug-use experience may limit and shape their subjective experience of craving cocaine and motivation for cocaine use. PMID- 15541778 TI - Temperament types are associated with weak self-construct, elevated distress and emotion-oriented coping in schizophrenia: evidence for a complex vulnerability marker? AB - Knowledge concerning temperament factors involved in vulnerability to schizophrenia is limited. We hypothesized that temperament and self-variables (emotional distress, coping styles, self-efficacy and self-esteem) might present a complex trait marker for underlying vulnerability to schizophrenia. We sought to (1) assess temperament dimensions and types in schizophrenia patients and healthy controls using the Tridimensional Personality Questionnaire (TPQ), and (2) explore their association with symptom dimensions, emotional distress, coping styles, self-constructs, demographic and background variables. We evaluated 90 consecutively recruited DSM-IV schizophrenia patients and 136 healthy controls matched for gender and age. We found that the harm avoidance (HA) factor was higher, while reward dependence (RD) was lower in schizophrenia patients than in healthy controls. Relationships of negative symptoms to novelty seeking (NS) and general psychopathology with both NS and HA show a confounding relation to self variables. TPQ temperament types were defined by dichotomization into high and low according to medians of the three TPQ temperament dimensions. The odds ratios for the HA and HA/NS temperament types were significantly higher, while the NS/RD type was lower in schizophrenia patients than in healthy controls. HA/NS, HA/RD and high-HA/NS/RD types revealed higher scores for emotional distress, emotion oriented coping and lower scores on self-constructs. No links were found between temperament types and schizophrenia symptom dimensions, task and avoidance oriented coping, or demographic and background variables. Thus, our findings strengthen the hypothesis that temperament types, when associated with elevated emotional distress, emotion-oriented coping and weak self-constructs, might represent a complex trait marker for underlying vulnerability to schizophrenia. PMID- 15541779 TI - Nicotine use in schizophrenia and disinhibition. AB - The role of personality in nicotine use has been little studied in patients with schizophrenia. The objectives of the present study were to compare impulsivity, sensation seeking and anhedonia in a group of schizophrenic patients with and without current cigarette smoking. One hundred patients with schizophrenia or schizoaffective disorder (DSM-III-R criteria) were assessed with the Positive and Negative Syndrome Scale, the Fagerstrom Scale, the Barratt Impulsivity Scale, the Zuckerman Seeking Sensation Scale, and the Chapman Physical Anhedonia Scale. Among these patients, 67% were current smokers. The mean scores for sensation seeking were higher in the group of smokers than in the group of nonsmokers. The differences were found exclusively with the "disinhibition" subscale. No significant difference was found for impulsivity and physical anhedonia mean scores. The scores on the Sensation Seeking Scale were especially low in nonsmokers. Cigarette smoking could influence disinhibition and possibly help to normalize inhibition in schizophrenic patients. PMID- 15541780 TI - Differences in facial expressions of four universal emotions. AB - The facial action coding system (FACS) was used to examine recognition rates in 105 healthy young men and women who viewed 128 facial expressions of posed and evoked happy, sad, angry and fearful emotions in color photographs balanced for gender and ethnicity of poser. Categorical analyses determined the specificity of individual action units for each emotion. Relationships between recognition rates for different emotions and action units were evaluated using a logistic regression model. Each emotion could be identified by a group of action units, characteristic to the emotion and distinct from other emotions. Characteristic happy expressions comprised raised inner eyebrows, tightened lower eyelid, raised cheeks, upper lip raised and lip corners turned upward. Recognition of happy faces was associated with cheek raise, lid tightening and outer brow raise. Characteristic sad expressions comprised furrowed eyebrow, opened mouth with upper lip being raised, lip corners stretched and turned down, and chin pulled up. Only brow lower and chin raise were associated with sad recognition. Characteristic anger expressions comprised lowered eyebrows, eyes wide open with tightened lower lid, lips exposing teeth and stretched lip corners. Recognition of angry faces was associated with lowered eyebrows, upper lid raise and lower lip depression. Characteristic fear expressions comprised eyes wide open, furrowed and raised eyebrows and stretched mouth. Recognition of fearful faces was most highly associated with upper lip raise and nostril dilation, although both occurred infrequently, and with inner brow raise and widened eyes. Comparisons are made with previous studies that used different facial stimuli. PMID- 15541781 TI - Generic and health-related quality of life in patients with seasonal and nonseasonal depression. AB - Although a relatively large body of research has now accumulated concerning the relationship between quality of life (QoL) and nonseasonal depression, there is a dearth of information about QoL in seasonal affective disorder (SAD). The aim of this study was to compare perceived levels of broad ('generic') and health related QoL in patients with seasonal and nonseasonal depression. Participants were 72 patients with SAD enrolled in an on-going multicentre study in Canada, and 72 patients with nonseasonal major depressive disorder (MDD) matched for severity of depression attending an outpatient psychiatric clinic in Vancouver, British Columbia. All participants completed two measures of QoL (the 20-item Medical Outcomes Study [MOS] Short-Form General Health Survey [SF-20] and the Quality of Life Enjoyment and Satisfaction Questionnaire [Q-LES-Q]) at baseline prior to treatment. The results of the study indicated that both generic and health-related QoL were compromised in patients with SAD compared with general population norms. For example, mean Q-LES-Q scores (range 0-100, where higher scores indicate better QoL) were 44%, compared with scores of 83% reported for the general population. Patients with nonseasonal depression showed significantly poorer functioning in several domains on the SF-20, but no significant differences in Q-LES-Q scores emerged. Perceived QoL is impaired in patients with SAD. Degree of impairment between seasonal and nonseasonal depressives is equivalent when assessed using the Q-LES-Q, but significant inter-group differences are apparent in SF-20 domain scores. Future research is required to determine whether perceived QoL is improved by treatment interventions for seasonal depression such as light therapy or antidepressant medication. PMID- 15541782 TI - The relationship between alexithymia, depression, and sleep complaints. AB - The study assessed in normal subjects the hypothesis of a specific association between alexithymia and poor sleep quality, taking into consideration the contribution of depression. Five hundred fifty-four university students (480 F and 74 M) filled out the Italian version of the 20-item Toronto Alexithymia Scale (TAS-20), the Center for Epidemiologic Studies Depression Scale (CES-D), and the Pittsburgh Sleep Quality Index (PSQI). TAS-20 scores were significantly correlated with many measures of self-rated poor sleep quality and also strongly correlated with depression scores. Any association between alexithymia and sleep complaints disappears when the contribution of depression is partialled out by multiple regressions, and only the well-known relationship between depression and impairment of sleep quality is confirmed. PMID- 15541783 TI - Lymphocyte subsets and lymphokine production in patients with melancholic versus nonmelancholic depression. AB - Several studies have reported immune changes during depression, but the results have not been fully consistent. Some of these changes could be related to the presence of melancholic features. A total of 42 depressed patients (melancholic [MEL] and nonmelancholic [non-MEL]) and 20 healthy controls participated in the study. We detected a higher CD4+ lymphocyte subset in MEL patients than in controls during the depressive state, which disappeared after clinical remission. We also found an increase in interleukin-2 (IL-2) production both in MEL and non MEL patients, but these values did not differ from control values after clinical remission. Some of these changes may be related to the melancholic characteristics of depression. PMID- 15541784 TI - Organizational strategy use in obsessive-compulsive disorder. AB - Patients with obsessive-compulsive disorder (OCD) have been demonstrated to be less likely to use spontaneously generated organizational strategies during verbal episodic memory and visuoconstruction tasks. However, whether this organizational deficit is generalizable to other areas of cognitive functioning has not been established. In the present study, we assessed whether adults with OCD are less likely to spontaneously generate organizational strategies during performance of an executive function test, the Self-Ordered Pointing Task (SOPT). Participants included 30 adults with OCD and 24 healthy controls. Groups did not differ with respect to the time to complete or number of errors made on the SOPT. Furthermore, group differences were not observed in the ability to generate organizational strategies or in the specific types of strategies employed to complete the SOPT. These findings indicate that a reduced use of organizational strategies in OCD is not present across all cognitive domains. PMID- 15541785 TI - Use of factor analysis to detect potential phenotypes in obsessive-compulsive disorder. AB - This study aimed to identify symptom dimensions in obsessive-compulsive disorder (OCD) in order to reveal distinct clinical phenotypes. Factor analysis of the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) checklist on item level was performed on data from 335 outpatients with primary OCD. The relationship of demographic and clinical characteristics to the resulting factor scores was examined. A principal component analysis identified the following five consistent symptom dimensions: (1) contamination and cleaning, (2) aggressive, sexual and religious obsessions, (3) somatic obsessions and checking, (4) symmetry and counting/arranging compulsions and (5) high-risk assessment and checking. We observed significant differences in sex distribution, age of onset, Y-BOCS scores and familial prevalence of OCD in relation to the symptom dimensions. These findings provide further evidence for distinct clinical phenotypes in OCD. PMID- 15541786 TI - Factor structure of the Zung Self-rating Depression Scale in first-year university students in Japan. AB - The Zung-Self-rating Depression Scale (SDS) was distributed to 28,588 first-year university students. Factor analysis using PROMAX rotation revealed three factors interpretable as affective, cognitive, and somatic symptoms. The confirmatory factor analysis showed a goodness-of-fit index of 0.976 and an adjusted goodness of-fit index of 0.967. The two sexes exhibited virtually the same factor structure. The result suggests that studies with this scale should use these three subscales rather than a total score. PMID- 15541787 TI - Heart rate variability in coronary artery disease patients with and without panic disorder. AB - Panic disorder (PD) and coronary artery disease (CAD) often co-occur, and CAD patients with comorbid PD suffer greater cardiovascular morbidity and mortality relative to CAD patients without PD. However, the mechanisms underlying these associations are still unknown. Reduced heart rate variability (HRV), a non invasive measure of cardiac autonomic modulation, is an important predictor of adverse cardiac events. Interestingly, reduced HRV has been observed in patients with panic-like anxiety and PD, as well as in various CAD populations. However, the extent to which HRV is altered in patients with both PD and CAD is unknown. This study evaluated HRV in 42 CAD patients with (n=20) and without (n=22) PD. Patients underwent 48-h electrocardiographic monitoring. Power spectral analysis of HRV indicated that CAD patients with PD exhibited significantly lower LF/HF ratios, which may reflect lower sympathetic modulation, compared with non-PD patients. Additionally, total power in PD patients was made up of a significantly higher proportion of HF power and a significantly lower proportion of VLF power than in non-PD patients. No other significant differences in HRV indices were observed. Results suggest that contrary to what has been observed in the majority of PD-only and CAD-only populations; patients with both PD and CAD appear to exhibit lower sympathetic modulation during ordinary daily life conditions. Though preliminary, these findings suggest that changes in HRV may not be the mechanism underlying greater cardiovascular morbidity and mortality among CAD patients with PD. PMID- 15541788 TI - Seasonality and suicide. PMID- 15541789 TI - Fc receptors in livestock species. AB - Many of the receptors for the Fc domain of immunoglobulins in cattle, sheep, pigs and horses have been cloned and characterized recently. This review summarises recent developments and relates them to the current understanding of the primary structure, cellular specificity and binding properties of Fc receptors (FcRs). Although there is an obvious overall similarity to their human and mouse counterparts, some Fc receptors in domestic animals are unusual, perhaps most notably the bovine Fcgamma2R, which although related to other mammalian FcgammaRs, belongs to a novel gene family and the porcine FcgammaRIIIA, which associates with a molecule that contains significant homology to the cathelin family of antimicrobial proteins. Accumulating data suggest the possibility of a different role for the FcRn in ruminants, which may secrete IgG onto the mucosal surfaces, rather than absorbing it, as was suggested by mouse studies. These differences may be linked to the diversity of immunoglobulin classes in different mammalian species, and may contribute to different immune functions. The observations made so far emphasize the importance of elucidating and analyzing the roles of these molecules within the immune system of each host animal, rather than inferring roles from conclusions made in human and mouse studies. A better understanding of Fc receptor expression on immune effector cells should help in developing new immunization protocols, while knowledge of the Fc receptors involved in immunoglobulin transport, especially in the mammary gland, may help to develop new products which could be used not only for veterinary purposes but perhaps also for human therapy. PMID- 15541790 TI - The proliferation inhibitory proteins p27(Kip1) and retinoblastoma are involved in the control of equine lymphocyte proliferation. AB - Observations in early equine pregnancy clearly reveal maternal immune recognition of and response to the presence of the conceptus. Nevertheless, both maternal cellular and humoral responses appear ineffective in destroying the developing placenta and fetus in early pregnancy. Our previous studies had shown that the pre-conditioned medium generated from the culture of equine invasive trophoblast inhibited mitogen-induced lymphocyte proliferation and the expression of cytokine messenger RNA in vitro. Those findings also suggested that lymphocytes might have been halted in the G0/G1 phase of the cell cycle. To characterize the cell cycle and the intracellular mechanisms involved in the inhibition of lymphocyte proliferation, equine peripheral blood lymphocytes were cultured in the presence or absence of pokeweed mitogen (PWM) in fresh medium, or in medium pre conditioned through cell culture of invasive trophoblast cells or fetal fibroblasts. Two-color flow cytometric analysis for bromodeoxyuridine (BrdU) incorporation by stimulated lymphocytes, and concomitant DNA staining with 7 amino-actinomycin D (7-AAD), indicated that a greater proportion of lymphocytes were found in the G0/G1 phase of the cell cycle when cultured in the invasive trophoblast cell pre-conditioned medium compared to controls. Analysis using carboxyfluorescein diacetate succinimidyl ester (CFSE) fluorescence intensity demonstrated that lymphocytes cultured in the presence of invasive trophoblast cell pre-conditioned medium had fewer cells going through division, but that those fewer cells sustained similar numbers of cell divisions as in control cultures. Hypophosphorylated retinoblastoma (Rb) protein expression was increased and p27Kip1 expression was maintained at higher levels in lymphocytes cultured in invasive trophoblast pre-conditioned medium compared to fresh medium. In agreement with these data, flow cytometric measurement of the Ki-67 protein expression in lymphocytes cultured in invasive trophoblast pre-conditioned medium was lower in comparison to controls. These findings suggest that the equine lymphocyte proliferation is at least partially regulated by the expression of proliferation inhibitory proteins such as p27Kip1 and hypophosphorylated Rb. These proteins seem to be important regulators of cell cycle transition between G1 and S phase in equine lymphocytes. PMID- 15541791 TI - Immune responses in rainbow trout Oncorhynchus mykiss induced by a potential probiotic bacteria Lactobacillus rhamnosus JCM 1136. AB - This study was undertaken to examine the effect of supplementing a suggested probiotic bacteria Lactobacillus rhamnosus JCM 1136 in feed on immune response and gut flora composition of rainbow trout Oncorhynchus mykiss. The probiotic bacteria were incorporated into a commercial feed to constitute two experimental diets containing either 10(9) or 10(11) colony forming unit of live bacteria/g of feed while a third diet without the bacterial supplement served as the control diet. The diets were offered to rainbow trout (75g average weight) in triplicate tanks for 30 days. Fish were sampled at 10, 20 and 30 days after commencement of the feeding trial to determine the proportion of the given probiont in the gut microflora composition and the nonspecific humoral and cellular immune responses on the 30th day. The relative proportion of the probiont increased with the feeding duration in the intestine, but not in the stomach. The proportion of L. rhamnosus in the stomach corresponded to the intake levels while no such relation existed in the intestine. The serum lysozyme and complement activities were significantly greater in fish fed the higher level of probiont compared with the control fish. The phagocytic activity of head kidney leucocytes also showed similar tendencies. These observations indicate the potential immuno-regulatory role of probiotic organisms in rainbow trout. PMID- 15541792 TI - Interferon-gamma response of PBMC indicates productive pseudorabies virus (PRV) infection in swine. AB - In Chinese Meishan/German Landrace cross-bred swine F2 generation interferon gamma (IFN-gamma) production by peripheral blood mononuclear cells (PBMC) was determined directly ex vivo at different time points after survival of a virulent pseudorabies virus (PRV) infection. This reactivity was compared with the reactivity of naive PBMC. Significant IFN-gamma production was determined in ELISA and ELISPOT only after in vitro PBMC re-stimulation with PRV and not with the closely related bovine herpesvirus BHV-1. The PRV-specific IFN-gamma secretion from re-stimulated PBMC showed high levels 6 days after infection, before the presence of serum antibodies, and it persisted at a high level over a 3 months period. The response of a group of eight piglets infected intranasally with PRV varied. Only two animals showed the expected typical fever response. PRV specific IFN-gamma production by PBMC clearly indicated that infection had occurred. Early significant IFN-gamma production by primed PBMC turned out to be a reliable and specific ex vivo marker for cellular response against productive PRV infection in swine before antibody formation. PMID- 15541793 TI - The effect of repeated tuberculin skin testing of cattle on immune responses and disease following experimental infection with Mycobacterium bovis. AB - The comparative intradermal skin test, in which a delayed type hypersensitivity (DTH) response to purified protein derivative of tuberculin (PPD) from Mycobacterium bovis and M. avium is assessed and compared, may be used repeatedly on non-infected animals on farms where bovine tuberculosis (TB) has occurred. A skin test is known to affect subsequent skin tests in infected animals. The reported study was to determine whether repeated skin testing prior to infection with M. bovis might affect the development of the comparative skin test and IFNgamma response subsequent to exposure to virulent M. bovis. The comparative intradermal skin test was applied to one group of six calves five times at 8-week intervals. These and six control calves were subsequently inoculated intratracheally with a dose of M. bovis that produced mild disease. The development of the DTH reaction, IFNgamma, IL-10 and proliferative responses were compared in the two groups of animals. No differences in IFNgamma, IL-10 and proliferative responses were seen between the two groups of calves prior to challenge. After infection with M. bovis no differences in the development of the DTH and IFNgamma responses to PPD were noted as a consequence of the repeated skin testing prior to challenge. No differences between the groups were evident when ESAT-6 was used as antigen and IFNgamma was assayed, although two animals that responded to PPD did not respond with ESAT-6. However, there did appear to be subtle effects of repeated skin testing on the immune response post-challenge that did not affect the diagnostic tests. After challenge control animals showed greater proliferative responses than animals given repeated skin tests prior to challenge, indicating that the procedure did have consequences for immune responses following infection. In both groups a marked reduction in the intensity of the skin test and in the number of animals that would be recognized as reactors was evident when animals were tested 15 weeks post-infection compared to their responses 8 weeks earlier that could have consequences for diagnosis of TB. An antibody response was not evident as a result of repeat skin testing prior to infection but was seen in both groups of calves following skin testing performed 7 weeks after infection. PMID- 15541794 TI - The effect of the tuberculin test and the consequences of a delay in blood culture on the sensitivity of a gamma-interferon assay for the detection of Mycobacterium bovis infection in cattle. AB - The strategic use of the gamma-interferon (IFN-gamma) assay (Bovigam) can provide a means for the early identification of Mycobacterium bovis infected cattle, thus ensuring their removal from an infected herd. It has been reported that performance of the test can be influenced by various factors including a recent tuberculin skin test and the length of delay between collection and processing of blood samples. In this study, single intradermal comparative tuberculin test (SICTT) reactor and non-reactor cattle were recruited from herds infected with M. bovis and grouped according to their SICTT responses. Group 1 comprised reactor cattle selected on the basis of their SICTT response to PPD-bovine (purified protein derivative of tuberculin) exceeding that of PPD-avian by at least 12mm. Group 2 animals were selected from herds undergoing routine surveillance for bovine tuberculosis and contained standard SICTT reactor cattle (PPD-bovine exceeding that of PPD-avian by at least 4mm) and non-reactors. We investigated the effects of the SICTT on the assay results by measuring the in vitro IFN-gamma responses of Group 1 reactor cattle at time intervals pre- and post-skin test. No significant differences were measured in the IFN-gamma responses of the reactor animals to PPD-bovine and PPD-avian for up to 65 days. To investigate if a delay in processing of blood affected the performance of the assay, we compared results using duplicate blood samples from Group 1 and Group 2 cattle stimulated with PPD antigen at 8h and at 24h after collection. In both groups of animals the mean optical density (OD) values of the assay at 24h post-collection were significantly lower than those at 8h. Our results demonstrated that a delay in processing of the blood samples from cattle subjected to routine surveillance could significantly impact on the outcome of the IFN-gamma assay resulting in a change of the IFN-gamma status of the animals. PMID- 15541795 TI - Bioactivity and secretion of interleukin-18 (IL-18) generated by equine and feline IL-18 expression constructs. AB - Interleukin 18 (IL-18) is a cytokine capable of induction of IFNgamma, granulocyte monocyte-colony stimulating factor (GM-CSF), TNFalpha and IL-1 in immunocompetent cells. Equine and feline plasmid vectors expressing pro-IL-18, mature IL-18 and IL-18 fused to a synthetic signal sequence from human IL-1beta receptor antagonist protein (ILRAP), ILRAP-IL-18, have been generated. In vitro protein expression of these constructs was compared by Western blot analysis. These data demonstrated that ILRAP-IL-18 protein was secreted readily from transfected chinese hamster ovary (CHO) cells. A simple bioassay for human IL-18 was recently described using human myelomonocytic KG-1 cells, which produce human IFNgamma in response to human IL-18 in a dose dependent manner (Konishi et al., 1997). We demonstrated bioactivity of equine and feline IL-18 protein in transfection products of CHO cells using this assay. Bioactivity of ILRAP-IL-18 protein was demonstrated in the culture medium of transfected CHO cells. These data imply that the ILRAP-IL-18 construct shows potential for use in vivo, where cell secretion of protein is crucial. PMID- 15541796 TI - Characterization of a newly established nonproducer lymphoma cell line for feline leukemia virus. AB - A feline lymphoblastoid cell line (KO-1) was established from a 5-year-old neutered female cat with naturally occurring thymic lymphoma. KO-1 cells had a rearrangement of T-cell receptor beta-chain gene and a germ-line configuration of immunoglobulin heavy chain gene, however, they were devoid of T-cell-specific surface phenotype. Cytogenetically, KO-1 cells showed a hyperploidy (2n = 41) due to the trisomy of B2, F2 and X chromosomes. Although KO-1 cells were shown to be clonally expanded cells integrated with feline leukemia virus (FeLV) proviruses and expressed its structural proteins in their cytoplasm, they did not produce virus particles as shown by transmission electron microscopy and the absence of the viral protein and reverse transcriptase activity in the culture supernatant. The present study showed that the KO-1 cell line established here was a feline T cell lymphoma cell line having a unique characteristic as an FeLV nonproducer. PMID- 15541797 TI - Identification of CpG oligodeoxynucleotide sequences that induce IFN-gamma production in canine peripheral blood mononuclear cells. AB - Oligodeoxynucleotides containing the cytosine-phosphate-guanine (CpG) motif (CpG ODNs) have been shown to induce T(H)1 immune responses in animals. Since the sequences of CpG-ODNs that induce T(H)1 responses are considered to vary among animal species, it is necessary to identify effective CpG-ODNs in each animal. In order to identify the sequences of CpG-ODNs that induce T(H)1 responses in dogs, mRNA expression and protein production of IFN-gamma were examined in peripheral blood mononuclear cells (PBMCs) from healthy dogs treated with 11 kinds of synthetic CpG-ODNs. One of the 11 CpG-ODNs (No. 2 CpG-ODN, 5' GGTGCATCGATGCAGGGGGG-3') was shown to significantly increase mRNA expression and protein production of IFN-gamma in canine PBMCs in a manner dependent on the sequence of the CpG motif. This CpG-ODN also enhanced the expression of IL-12 p40 mRNA in canine PBMCs, whereas expression of IL-12 p35, IL-18, and IL-4 mRNAs was not induced by this CpG-ODN. These results indicate that this CpG-ODN was able to produce IFN-gamma by induction of T(H)1-skewed immune response in dogs. CpG-ODNs may be useful for inducing prophylactic and therapeutic immunity against allergic diseases, viral infection, and tumors in dogs. PMID- 15541798 TI - Interactions in biofilms between Listeria monocytogenes and resident microorganisms from food industry premises. AB - Twenty nine bacterial strains were grown as binary culture biofilms with Listeria monocytogenes to assess their influence on the settlement of the latter on stainless steel coupons. Most of the strains had been isolated from food processing plants after cleaning and disinfection and were tentatively identified by the APILAB Plus 3.3.3 database (bioMerieux). Sixteen of them decreased L. monocytogenes biofilm colony forming units (CFU) counts. Three strains, Bacillus sp. CCL 9 an unidentified Gram-positive strain CCL 59 and Pseudomonas fluorescens E9. 1, led to a 3-log difference in CFU counts between the pure L. monocytogenes biofilms and the mixed biofilms. Eleven strains had no effect and only four, Kocuria varians CCL 73, Staphylococcus capitis CCL 54, Stenotrophomonas maltophilia CCL 47 and Comamonas testosteroni CCL 24, had a positive effect, with a 0.5- to 1.0-log increase in the L. monocytogenes biofilm CFU counts. On its own, L. monocytogenes settled as single cells, but in binary biofilms, different spatial arrangements were observed: (i) with K. varians CCL 73, K. varians CCL 56 and S. capitis CCL 54, L. monocytogenes cells gathered around the microcolonies of the partner strain; (ii) with the two Gram-negative strains, C. testosteroni CCL 24 and CCL 25, L. monocytogenes cells formed its own microcolonies. No link could be found between the exopolysaccharide production capacity of the bacterial strains in pure-culture biofilms and their effect on the L. monocytogenes population in mixed biofilms. With one strain, C. testosteroni CCL 24, adding filter-sterilized supernatant from a pure-culture biofilm to a pure culture of L. monocytogenes increased the number of L. monocytogenes cells adhering to the stainless steel coupons and forming microcolonies. This study suggests that the "house flora" can have a strong effect on the likelihood of finding L. monocytogenes on inert surfaces. PMID- 15541799 TI - Purification, characterization and amino acid sequencing of divergicin M35: a novel class IIa bacteriocin produced by Carnobacterium divergens M35. AB - Carnobacterium divergens M35, isolated from a commercial sample of frozen smoked mussels, produces a new bacteriocin, divergicin M35, a class IIa bacteriocin. Divergicin M35 is sensitive to pronase-E, alpha-chymotrypsin and proteinase K, but not to trypsin and withstands thermal treatments up to 121 degrees C for 30 min. Divergicin M35 was extracted from the culture supernatant of C. divergens M35 using an SP-Sepharose cation-exchange column, desalted and purified on a C18 Sep-Pack column and further purified by reverse phase-high pressure liquid chromatography. This procedure allowed the recovery of 10% of the bacteriocin present in the culture supernatant with purity higher than 99%. Divergicin M35 had a molecular mass of 4518.75 Da as determined by mass spectrometry, a pI value of 8.3 and positive net charge (+3). The amino acid sequence of divergicin M35 was found to consist of 43 amino acid with four cysteine residues (Cys10, 15, 25, 43) and showed 80.5% homology with divercin V41 (80.5%) and 80.0% with bavaricin MN. Divergicin M35 showed powerful antilisterial activity, especially against Listeria monocytogenes and was also active against carnobacteria but not against strains of Lactococcus, Lactobacillus, Enterococcus, Bifidobacteria and Escherichia. Divergicin M35 production began in late exponential phase and reached a maximum activity of 65,000 AU/ml in early stationary phase. Initial broth pH, Tween 80 and acetate did not affect C. divergens M35 growth or divergicin production. This bacteriocin may be a potential tool for inhibiting L. monocytogenes in seafood products that do not usually undergo an adequate heat treatment. PMID- 15541800 TI - Application of extended single-reaction multiplex polymerase chain reaction for toxin typing of Staphylococcus aureus isolates in South Korea. AB - The extended single-reaction multiplex PCR (esr-mPCR) developed in this study to detect staphylococcal enterotoxins (SEs), including SEA, SEB, SEC, SED, SEE, SEH, SEI, and SEJ, requires fewer sets of primers than other conventional multiplex PCRs and can be used to detect newly identified staphylococcal enterotoxins SEs more readily. Esr-mPCR analysis of 141 isolates of Staphylococcus aureus obtained from abattoir and livestock product samples revealed that 27 of the S. aureus isolates were toxigenic, and two were 2 multitoxigenic isolates. The most prevalent SE type was SEI followed by SEA and SEH. In addition, we investigated the clonal relatedness of toxigenic S. aureus isolates by arbitrarily primed PCR (AP-PCR). AP-PCR analysis of toxigenic S. aureus isolates revealed that the discriminatory power of AP-PCR was 9 (D=0.81), 8 (D=0.77), and 10 types (D=0.83) with primers AP1, ERIC2, and AP7, respectively. The combination of three each AP PCR result could rearrange toxigenic S. aureus isolates into 10 types and five subtypes, with the D-value of 0.92. Interestingly, our data showed that toxigenic S. aureus isolates from different sources had different fingerprinting patterns although some of them carried the same types of SE genes. These data suggest that combinations of esr-mPCR and AP-PCR can provide a powerful approach for epidemiological investigation of toxigenic S. aureus isolates. PMID- 15541801 TI - Numbers and strains of lactobacilli in some probiotic products. AB - The numbers and types of Lactobacillus strains in probiotic feed or food products for sale in Europe were compared with the information stated on the product labels. The labels of all 10 products gave information on the genus and species of Lactobacillus, and seven gave information on the numbers of a specified strain(s). Lactobacilli were recovered using a recently validated method, developed as part of the European Community Project SMT4 CT98-2235 for the official control of probiotic microorganisms used as feed additives. Polymerase chain reaction (PCR)-based methods were used to assess the accuracy of labelling with regard to genus and species, and pulsed-field gel electrophoresis (PFGE) was used to identify strains. Five products were mislabelled with respect to the numbers and three with respect to species of lactobacilli. In four cases, the specified strains were not detected. Four fermented milks sold under three trademarks contained the same strain, which was named differently on each label. As safety and functionality of probiotics are strain dependent, these results demonstrated the need to control lactobacilli present in commercially probiotic human food products, not only at the species but also at the strain level, to ensure their quality and protect the consumer. The usefulness of the official methods developed for animal feed additives to identify lactobacilli in probiotic foods for humans was demonstrated. PMID- 15541802 TI - Metabolism of extracellular inositol hexaphosphate (phytate) by Saccharomyces cerevisiae. AB - Iron and zinc deficiencies are global problems, frequently leading to severe illness in vulnerable human populations. Addition of phytases can improve the bioavailability of iron and zinc in food. Saccharomyces cerevisiae would be an ideal candidate as a bioavailability improving food additive if it demonstrates significant phytase activity. The purpose of the paper was to study yeast phytase activity to obtain information required to improve strains. All yeasts tested readily degraded extracellular inositol hexaphosphate (phytate; IP6) in media with IP6 as the sole phosphorous source. Phosphate (Pi) addition yielded repression consistent with the PHO system. However, repression of IP6-degrading enzymes was not only dependent on level of Pi, but also on pH and medium composition. In complex medium, containing Pi at a concentration previously suggested to yield full repression of the secretory acid phosphatases (SAPs; e.g., [Mol. Biol. Cell 11 (2000) 4309]), and at relatively high pH, repression of phytate-degrading enzymes was weak. The capacity to degrade phytate, irrespective of Pi addition or not, was highest at the pH most distant from the pH optimum of the SAPs [Microbiol. Res. 151 (1996) 291], suggesting that expression rather than enzyme activity was affected by pH. In synthetic medium, repression was strong and pH-independent (no IP6 degradation within the range tested). The distinct difference between media shows that, in addition to known regulatory role of Pi for the PHO system, additional factors may be involved. Using a deletion strain, we further demonstrate that the main secretory acid phosphatase Pho5p is not essential for intact phytate-degrading capacity and growth without Pi, neither is Pho3p. However, when constitutively overexpressing PHO5 an increased net phytase activity was obtained, in repressing and non-repressing conditions. This proves that, although redundant in a wild type, Pho5p can catalyze hydrolysis of IP6 and that at least one more enzyme is capable of effective hydrolysis of IP6 (sufficient to provide the cell with phosphorous at a rate yielding maximum growth). Finally, a bread dough experiment showed that the typical concentrations of Pi during leavening exceed levels shown to repress phytate degradation by a wild-type S. cerevisiae. PMID- 15541803 TI - Proximate sources of bacteria on boneless loins prepared from routinely processed and detained carcasses at a pork packing plant. AB - Microbiological samples were obtained by swabbing detained and routinely processed pig carcasses before and after cooling, and sides, loin portions and loin cuts at various stages of the carcass breaking process. Aerobes, coliforms and Escherichia coli were enumerated in each sample. All three groups of bacteria were more numerous on detained than on routinely processed carcasses. Both trimming and cooling reduced the numbers of E. coli but not the numbers of aerobes on detained carcasses. After cooling, the log mean number of aerobes and E. coli on detained carcasses were each about 0.5 log unit more than the log mean numbers on routinely processed carcasses, but numbers of coliforms on the two types of carcass were similar. There were small increases in the numbers of coliforms and E. coli on carcasses during their movement from the cooler to the breaking facility. The numbers of bacteria on the meat apparently did not increase during the carcass-breaking process, although bacteria were redistributed on the product. Despite that, substantial numbers of bacteria were recovered from parts that do not contact food in cleaned conveying equipment used for carcass breaking. However, those bacteria included few coliforms and no E. coli. These findings suggest that the contamination of meat with E. coli from persistent detritus in carcass breaking equipment, such as has been found to occur at beef packing plants, may be prevented when carcass-breaking equipment and facilities are dried after cleaning, and wetting of equipment during processing is avoided. PMID- 15541804 TI - Detection of noroviruses in raspberries associated with a gastroenteritis outbreak. AB - Following an acute foodborne gastroenteritis outbreak in southern Sweden, stool specimens from five of nine ill patients were found positive for norovirus using reverse transcriptase polymerase chain reaction. Epidemiological data pointed to raspberry cakes as the source of the outbreak. Using a combination of generic and patient-specific primers and novel food analysis methodology (with extraction efficiency control and inhibitor removal), norovirus strains from two different genogroups were directly identified in the contaminated raspberries. PMID- 15541805 TI - Survival of Campylobacter jejuni on various fresh produce. AB - The survival of seven human and two chicken Campylobacter jejuni strains, with known Penner heat-stable (HS) serotypes and pulsed-field gel electrophoresis (PFGE) genotypes, was investigated on fresh-cut iceberg lettuce. In addition, the survival of four selected C. jejuni strains was assessed on cantaloupe pieces, cucumber slices, grated carrot and strawberries. Fresh produce was inoculated with 10(5) to 10(7) colony-forming units (CFU) of C. jejuni per gram, and the bacterium was enumerated using standard procedures after sample storage at 7 and 21 degrees C for 24, 48 and 72 h. The absolute values of the slopes (death rates) of the survival curves (log(10) CFU/g versus time) were calculated and compared. At 7 degrees C, the mean death rates (day(-1)) were 0.44 on cantaloupe, 0.41 on cucumber slices, 0.43 on grated carrot, 0.59 on iceberg lettuce and 1.02 on strawberries. The corresponding death rates (day(-1)) at 21 degrees C were 1.52, 1.55, 2.61, 1.39 and 8.74. The death rate of C. jejuni on strawberries (pH 3.4) was significantly (P<0.05) higher than on other produce. Moreover, the death rate at 21 degrees C as compared with 7 degrees C was significantly higher (P<0.05). Minor differences were observed in the survival of different C. jejuni strains. Our results suggest that after contamination of fresh produce, including strawberries, C. jejuni may survive sufficiently long to pose a risk to the consumer. PMID- 15541806 TI - Molecular quantification of lactic acid bacteria in fermented milk products using real-time quantitative PCR. AB - Real-time quantitative PCR assays were developed for the absolute quantification of lactic acid bacteria (LAB) (Streptococcus thermophilus, Lactobacillus delbrueckii, L. casei, L. paracasei, L. rhamnosus, L. acidophilus and L. johnsonii) in fermented milk products. The results of molecular quantification and classic bacterial enumeration did not differ significantly with respect to S. thermophilus and the species of the L. casei group which were detected in the six commercial fermented products tested, thus showing that DNA extraction was efficient and that genomic DNA solutions were free of PCR inhibitors. For L. delbrueckii, the results of bacterial enumeration were generally lower by a factor 10 to 100 than those of PCR quantification, suggesting a loss of viability during storage of the dairy products at 1-8 degrees C for most of the strains in this species. Real-time quantitative assays enabled identification of the species of lactic acid bacterial strains initially present in commercial fermented milk products and their accurate quantification with a detection threshold of 10(3) cells per ml of product. PMID- 15541807 TI - Effects of modified atmosphere and vacuum packaging on microbiological and chemical properties of rainbow trout (Oncorynchus mykiss) fillets. AB - Microbial (psychrotrophic, mesophilic aerobic bacteria and Enterobacteriacae counts), and chemical analysis [pH, total volatile bases nitrogen (TVB-N), lipid oxidation (Thiobarbituric acid reactive substance, TBARS)] of rainbow trout (Oncorynchus mykiss) fillets in air (control), vacuum and modified atmosphere packaging (MAP) with various gas mixtures conditions at 4+/-1 degrees C were determined. The gas mixtures evaluated were 100% CO2, 2.5% O2+7.5% N2+90% CO2 and 30% O2+30% N2+40% CO2. Psychrotrophic bacteria count was above 1 x 10(7) cfu/g on the 12th day in 100% CO2. However; mesophilic bacteria count was below 1 x 10(6) cfu/g at the end of the 14-day storage period. Enterobacteriaceae count was significantly lower in samples packaged with MAP. Lipid oxidation increased rapidly after 6 days of storage in the samples containing 30% O2. While minimum TBARS values were recorded in fillets containing 100% CO2 and vacuumed fillets, the lowest TVB-N values were obtained in fillets with 100% CO2. PMID- 15541808 TI - Factors affecting the antilisterial effects of nisin in milk. AB - The ability of Listeria monocytogenes to proliferate in milk and the antilisterial activities of nisin are well documented. Although milk fat was reported to reduce the antimicrobial activities of nisin, there is little information on the influence of milk fat on the antilisterial activities of nisin in refrigerated milk, and whether pasteurization and homogenization influence these activities. Fresh, pasteurized, and homogenized milk samples (0.1%, 2.0%, and 3.5% fat) were treated with nisin (0-500 IU/ml) and challenged with 10(4) CFU/ml L. monocytogenes strain Scott A. The organism was most sensitive to nisin in skim milk, showing rapid decline in cell numbers to <10 CFU/ml after 12 days at 5 degrees C following treatment with 250 IU/ml. An initial decline in cell numbers in 2% and whole milk was followed by regrowth of the organism. Loss of the antilisterial effects of nisin was confirmed in homogenized whole milk, whether raw or pasteurized, but not in raw or pasteurized whole milk that was not homogenized. Tween 80, a nonionic emulsifier, partially counteracted the loss of the antilisterial activity of nisin, whereas lecithin, an anionic emulsifier, had no effect. These results demonstrate that the chemical composition and treatment of foods may play an important role in the antilisterial effects of nisin. PMID- 15541809 TI - Absence ochratoxin A in soy sauce. AB - A method is described for the determination of ochratoxin A (OTA) in soy sauce using phosphate-buffered saline (PBS) extraction, an immunoaffinity clean-up, a liquid chromatographic determination with fluorescence detection (LC-FD) and confirmation with LC-FD after methylation of OTA. Recoveries of OTA spiked to soy sauce samples at 0.25 ng/ml level were 90% with relative standard deviations of 4%. The limit of detection was 0.01 ng/ml for OTA using the proposed method. Furthermore, the proposed method was applied to 60 soy sauce samples from China and Japan and none of them were found to contain OTA. PMID- 15541810 TI - Lung cancer mortality reduction after installation of tap-water supply system in an arseniasis-endemic area in Southwestern Taiwan. AB - Arsenic has been well documented as the major risk factor for blackfoot disease (BFD), a unique peripheral vascular disease (PVD) that was endemic in the Southwestern coast of Taiwan, where residents had used high-arsenic artesian well water for more than 50 years. Long-term arsenic exposure has also been reported to associated with lung cancer mortality in a dose-response relationship. A tap water supply system was implemented in the early 1960s in the BFD-endemic areas. Artesian well water was no longer used for drinking and cooking after the mid 1970s. The objective of this study is to examine whether lung cancer mortality decreased after the improvement of drinking water supply system through elimination of arsenic exposure from artesian well water. Standardized mortality ratios (SMRs) for lung cancer were calculated for the BFD-endemic area for the years 1971-2000. The study results show that mortality from lung cancer declined gradually after the improvement of drinking water supply system to eliminate arsenic exposure from artesian well water. Based on the reversibility criterion, the association between arsenic exposure and lung cancer mortality is likely to be causal. PMID- 15541811 TI - Molecular analysis of the TSC1 gene in adenocarcinoma of the lung. AB - We previously reported that loss of heterozygosity (LOH) in tuberous sclerosis 1 (TSC1)-gene-associated region on chromosome 9q34, was frequently observed in adenocarcinoma (AC) of the lung and its putative precursor lesion, atypical adenomatous hyperplasia (AAH). Some novel tumor suppressor gene for AC of the lung may be present in this region, and the TSC1 gene located on chromosome 9q34 is one candidate. The aim of this study was to determine whether the TSC1 gene itself acts as a tumor suppressor gene in the pathogenesis of AC of the lung. DNA extracted from frozen tumor tissue was used to screen 47 ACs of the lung for the presence of mutations in 21 coding exons of the TSC1 gene. When using these bulk tissue specimens, three different types of mutations were detected in five ACs. These positive samples were then submitted to laser capture microdissection to selectively collect tumor cells, DNA extraction and finally analysis of LOH and mutations of the TSC1 gene. Mutations were detected in three ACs, and LOH was detected in another AC. No mutations or LOH was detected in the other AC. Loss of heterozygosity and mutations were not detected simultaneously in the same tumor. We concluded that the TSC1 gene itself does not act as a tumor suppressor gene in the pathogenesis of AC of the lung in accordance with classical Knudsen's two-hit hypothesis. However, the frequent LOH in AC and AAH shown in our previous studies indicate the following possibility in the etiology of AC of the lung: the presence of a novel tumor suppressor gene close to the TSC1 gene, aberrant promoter methylation, or haploid insufficiency of the TSC1 gene. PMID- 15541812 TI - The multi-targeted kinase inhibitor SU5416 inhibits small cell lung cancer growth and angiogenesis, in part by blocking Kit-mediated VEGF expression. AB - SU5416 is a multi-targeted kinase inhibitor that potentially has the ability to directly block tumor growth by inhibiting Kit signaling, as well as blocking angiogenesis by inhibiting vascular endothelial growth factor receptor (VEGFR) signaling. Previous work has demonstrated that SU5416 efficiently blocks Kit mediated growth of small cell lung cancer (SCLC) in vitro. To determine the drug's effect on in vivo growth of SCLC, we studied its activity, alone and in combination with carboplatin, in chemotherapy-resistant H526, and chemotherapy sensitive H209 murine xenograft models. SU5416 efficiently inhibited Kit activity in vivo when administered on a twice-weekly schedule. When administered over a 3 week period to animals bearing established tumors, it inhibited growth by at least 70%. It was at least as effective as carboplatin in suppressing growth of H526 xenografts. However, the combination with carboplatin was not superior to the most active single agent in either xenograft model at the doses and schedule utilized. SU5416 clearly inhibited growth in part by inhibiting angiogenesis, with microvessel density dropping by approximately 50% in treated xenografts. In addition to the recognized mechanism of inhibition of VEGFR, we uncovered a novel mechanism of angiogenesis suppression by demonstrating reduced VEGF expression in SU5416-treated xenografts. In vitro, stem cell factor treatment of the H526 cell line enhanced expression of VEGF, which was efficiently blocked with SU5416. Thus, we have demonstrated that SU5416 can inhibit SCLC growth by directly inhibiting tumor cell proliferation and by inhibiting angiogenesis, in part by inhibiting Kit-mediated VEGF expression. These data suggest that kinase inhibitors that target both Kit and VEGFR could play an important role in the treatment of SCLC, as well as other malignancies that express Kit. PMID- 15541813 TI - VEGF gene sequence variation defines VEGF gene expression status and angiogenic activity in non-small cell lung cancer. AB - Different vascular endothelial growth factor (VEGF) gene polymorphisms have been shown to result in different VEGF gene responsiveness to various stimuli and different capacity for VEGF protein production. In the present study, we examined four VEGF gene polymorphisms in thirty-six individuals with non-small cell lung cancer (NSCLC). Gene polymorphisms were correlated with the VEGF protein expression in cancer cells and the tumor angiogenic activity. The -2578C/C, 634G/G and -1154A/A and G/A alleles in the VEGF gene were linked with low VEGF expression, while the -2578C/A, the -634 G/C and the -1154G/G alleles were linked with high VEGF expression. Tumors with -2578C/C had a significantly lower vascular density (VD) compared to the -2578C C/A. Similarly, cases with the 634G/G VEGF polymorphism had a singinificanltly lower vascular density compared to the combined C/C and G/C groups. In addition, the -1154A/A polymorphism seemed to relate with poor vaccularization but the difference did not reach significance. It is concluded that inherited VEGF sequence variations, which characterize the tumor genome itself, are strong determinants of the molecular VEGF and VEGF-downstream phenotype of NSCLC. The large variation in angiogenicity between tumors of similar histologic morphology emerges as a consequence of the 'parental' VEGF gene ability to produce VEGF. PMID- 15541814 TI - Expression of survivin mRNA and livin mRNA in non-small-cell lung cancer. AB - It has been suggested that suppression of apoptosis may contribute to the development and progression of cancer. Anti-apoptotic survivin and livin genes are highly expressed in cancer cells and transformed cells, but show little or no expression in normal differentiated tissues. However, there are no available data concerning livin expression in non-small-cell lung cancer (NSCLC). We therefore measured livin mRNA and survivin mRNA expression in 38 NSCLC cancer samples and 15 paired non-cancerous lung tissue samples using a quantitative reverse transcription-polymerase chain reaction (RT-PCR). While both mRNAs showed higher expression in cancers than non-cancerous tissues (P < 0.001), livin mRNA and survivin mRNA expression did not correlate with one another. When a cut-off value for positivity was set at the mean + S.D. for expression values in non-cancerous tissues, positivity rates for livin mRNA and survivin mRNA expression were 76.3% (29 of 38) and 36.8% (14 of 38) in lung cancers and 6.7% (1 of 15) and 0% (0 of 15), respectively, in paired non-cancerous lung tissue samples. Livin mRNA and survivin mRNA expression in tumors were up-regulated in 23 of 31 (74.2%) early stage NSCLC patients and 11 of 31 (35.5%), respectively. Expression of both mRNAs in tumors varied independently of tumor histology. These results support the possibility that the livin gene may play a role in NSCLC development and increased expression of livin mRNA may serve as a new target for lung cancer treatment as well as survivin. PMID- 15541815 TI - Clinicopathological significance of aberrant methylation of RARbeta2 at 3p24, RASSF1A at 3p21.3, and FHIT at 3p14.2 in patients with non-small cell lung cancer. AB - We investigated the clinicopathological significance of aberrant methylation of the retinoic acid receptor-beta2 (RARbeta2), RAS association domain family 1A (RASSF1A) and fragile histidine triad (FHIT) genes located on choromosome 3p in 120 patients with primary non-small cell lung cancer (NSCLC) by a methylation specific PCR method. Aberrant methylation of these was detected in 31 (26%), 35 (29%) and 43 (36%) tumors, respectively. There was no correlation with the methylation status of any of the genes. RARbeta2 methylation was more frequently observed in patients with a smoking history (19 of 61, 31%) than in patients without one (3 of 29, 10%, P = 0.0373). RARbeta2 methylation was also preferentially observed in advanced stage NSCLC (12 of 71 (17%) in stage I, 5 of 15 (33%) in stage II, 11 of 24 (46%) in stage III, and 3 of 8 (38%) in stage IV, P = 0.0057 (stage I versus II, III,and IV)). FHIT methylation was predominantly detected in tumors with vascular invasion (21 of 44, 48%, P = 0.0703) or lymphatic permeation (28 of 59, 47%, P = 0.0115). RASSF1A methylation was more frequently observed in adenocarcinomas (28 of 72, 39%) than in squamous cell carcinomas (6 of 45, 13%, P = 0.0033). These results indicate that aberrant methylation of the candidate tumor suppressor genes on 3p plays a respective role in the pathogenesis of NSCLC. PMID- 15541816 TI - Translating biomarkers into clinical practice: prognostic implications of cyclophilin A and macrophage migratory inhibitory factor identified from protein expression profiles in non-small cell lung cancer. AB - Biomarkers have the potential to significantly change diagnostic strategies and influence therapeutic management. We developed a MALDI-TOF protein expression profiling platform for biomarker discovery and a proof-of-principle study identified two proteins, cyclophilin A (CyPA) and macrophage migration inhibitory factor (MIF), that were overexpressed in non-small cell lung cancer (NSCLC). The current study focused on evaluating the potential of CyPA and MIF as prognostic markers in patients with a new diagnosis of lung cancer for rapid translation into clinical practice. Two hundred and thirty-four primary NSCLC specimens reflecting a broad range of histologies and stages were examined for CyPA and MIF reactivity by tissue microarray immunohistochemistry (TMA-IHC). The percent tumor cell reactivity, staining intensity and a composite staining score were compared with overall patient survival by Kaplan-Meier curves, log rank test and Cox model statistics. Although both proteins were overexpressed in most NSCLC tumors, neither CypA nor MIF showed a correlation with outcome. This pilot project approach can expedite integration of newly discovered biomarkers into clinical practice, with the goal of improving stratification of patients into appropriate treatment regimens. While both proteins considered in this study were overexpressed in the vast majority of NSCLCs, they were not found to be of prognostic significance. PMID- 15541817 TI - Prognostic value of tumor size in non-small cell lung cancer larger than five centimeters in diameter. AB - OBJECTIVES: The tumor size is an important prognostic factor in non-small cell lung cancer (NSCLC). However, a criterion for tumor size has remained unchanged for the last 25 years in TNM classification. The purpose of this study was to determine the relationship between tumor size and survival in patients with non small cell lung cancer. METHOD: Of 550 consecutive patients who underwent operation for NSCLC between 1994 and 1998, we reviewed the medical record of 509 patients. There were 470 men and 39 women. Survivals were compared according to groups of tumor size (Group I: < or = 3 cm, Group II: 3.1-4 cm, Group III: 4.1-5 cm, Group IV: >5 cm, Group A (II + III): 3.1-5 cm, and Group B (IV): >5 cm). The Cox proportional hazard model was used with same variables. RESULTS: The univariate analysis showed that poor pulmonary function test (P < 0.05), pneumonectomy (P < 0.05), limited resection (P < 0.001), tumor size larger than 5 cm (P = 0.006), T factor (P < 0.05), N status (P < 0.001), and advanced staged of disease (P < 0.001) were all significant prognostic factors. Further comparison of survival between tumor size groups demonstrated a significantly poor prognosis for larger tumors. There was a statistically significant difference between Group A (3.1-5 cm) and Group B (> 5 cm), with 5 years survivals of 45.9% and 31.4%, respectively (P = 0.005). CONCLUSION: We emphasize that tumor size is an important prognostic factor in NSCLC. The 5 years survival with tumors larger than 5 cm (31.4%), is significantly less than the tumors 3.1-5 cm (45.9%) (P = 0.005). These initial results should strongly encourage additional studies in different countries on the interaction between tumor size and lung cancer characteristics to better clarity. In future revisions of the TNM classification, 5 cm may be a new threshold. PMID- 15541818 TI - High serum YKL-40 level in patients with small cell lung cancer is related to early death. AB - YKL-40, a growth factor for connective tissue cells, is secreted by cancer cells and macrophages. Elevated serum YKL-40 in patients with metastatic carcinoma has been associated with poor prognosis. We evaluated serum YKL-40 in 131 patients with small cell lung cancer (SCLC). Twenty-two percent of the patients with limited disease and 40% of the patients with extensive disease had elevated serum YKL-40. The median survival was 5.1 months for patients with elevated serum YKL 40 and 9.0 months for patients with normal serum YKL-40. Patients with elevated serum YKL-40 had increased hazard for death within the first 6 months after the start of chemotherapy compared to patients with normal serum YKL-40 (HR = 2.06, P = 0.009). Multivariate Cox analysis including routine prognostic variables showed that serum YKL-40 (P = 0.02) is independent of prognostic variables for survival within the first 6 months. Studies are needed to determine the function of YKL-40 in SCLC. PMID- 15541819 TI - Synchronous lesions detected by autofluorescence bronchoscopy in patients with high-grade preinvasive lesions and occult invasive squamous cell carcinoma of the proximal airways. AB - The cancerization field concept implies that lung cancer multicentricity may be a frequent event and primary studies using white-light bronchoscopy (WLB) have reported a high prevalence of multicentricity in patients with roentgenographically occult lung cancer. We have used autofluorescence bronchoscopy (AFB) to reassess the prevalence of synchronous lesions in patients referred for the staging and/or treatment of occult lesions initially detected during WLB. All the patients referred with high-grade preinvasive lesions (severe dysplasia, DYS S and carcinoma in situ, CIS) and occult invasive squamous cell carcinoma (CIV) of the bronchus initially detected during WLB at other centers, underwent AFB. Data were prospectively collected and retrospectively analyzed to assess the prevalence of synchronous occult lesions. From January 1996 to December 2001, 28 patients (26 males, 2 females; mean age: 65 +/- 11) were assessed. After re-evaluation, in two cases, the referred lesions corresponded only to metaplasia and were discarded from analysis. The 26 other patients were referred for 28 lesions (3 DYS S, 19 CIS and 6 CIV; 2 patients were referred with two synchronous lesions). AFB revealed, in these 26 patients, six additional lesions (1 DYS S, 4 CIS and 1 CIV). Multicentricity in this group, initially estimated to amount to 7% with WLB alone, raised to 23% by using AFB. The high prevalence of synchronous lesions in this series of patients with occult DYS S, CIS and occult CIV suggests that AFB may be a useful adjunct in the pretreatment evaluation. PMID- 15541820 TI - Pit-fall sign on computed tomography predicts pleural involvement and poor prognosis in non-small cell lung cancer. AB - Pleural involvement by a peripheral non-small cell lung cancer (NSCLC) is an important prognostic factor, but the accurate preoperative evaluation is sometimes difficult. We have already identified a feature on computed tomography (CT) that suggests a possible pleural involvement (pit-fall sign, i.e. multiple linear strands between the nodule and chest wall and/or inter-lobar fissure), and the clinical impact was assessed in the present study. A total of 103 consecutive peripheral NSCLC patients were prospectively examined, and the sensitivity, specificity, and accuracy of pit-fall sign in the diagnosis of pleural involvement were 83.3, 76.1, and 78.6%, respectively. The 5-year survival rate of pit-fall sign-positive patients was 46.5%, which was significantly lower than that of pit-fall sign-negative patients (68.9%; P = 0.044), and the prognostic significance was confirmed by a multivariate analysis. In conclusion, the pit fall sign on preoperative CT suggests a possible pleural involvement correlated with a poor prognosis. PMID- 15541821 TI - Magnetic resonance imaging and computed tomography in the diagnoses of brain metastases of lung cancer. AB - We evaluated the usefulness of double-dose (0.2 mmol/kg of gadoteridol) contrast enhanced magnetic resonance imaging (C-E MRI) in detecting brain metastases of lung cancer. We prospectively enrolled 134 patients with lung cancer who had no neurologic symptoms and who underwent a staging work-up. Patients were assigned to receive both contrast-enhanced computerized tomography (C-E CT) and double dose C-E MRI. Double-dose C-E MRI detected brain metastases in 19 patients, while C-E CT detected brain metastasis in only 12 of the 19 (P = 0.02). The 3-month survival rate for patients in double-dose C-E MRI group was found to be 2.06 times that of patients in a C-E CT group (P = 0.029), although the survival rate fell to 1.45 (P = 0.387) at 6 months. The results imply that double-dose C-E MRI changed the clinical stage of lung cancer patients. We concluded that double-dose C-E MRI improves the rate of detection of brain metastases during the initial staging of lung cancer. PMID- 15541822 TI - Randomized phase II feasibility study of combining the matrix metalloproteinase inhibitor BMS-275291 with paclitaxel plus carboplatin in advanced non-small cell lung cancer. AB - BACKGROUND: This randomized, double-blind, placebo-controlled study was designed to assess whether the addition of the matrix metalloproteinase (MMP) inhibitor BMS-275291 to combined paclitaxel and carboplatin chemotherapy had an adverse impact on expected tumor response or had significant toxicity, especially arthrotoxicity, in patients with advanced non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Seventy-five chemotherapy-naive patients with stage IIIB-IV NSCLC were randomly assigned to BMS-275291 or placebo. All patients received paclitaxel 200mg/m(2) as a continuous 3-hour infusion followed by carboplatin calculated using the Calvert formula for a target AUC of 6 mg / (ml min), every 21 days for a maximum of eight cycles. BMS-275291 or placebo was administered on an outpatient basis at a daily oral dosage of 1200 mg. RESULTS: All 75 patients were evaluable for toxicity and 65 (86.7%) for response. Drug-related arthrotoxicity > or =grade 2 occurred in 12 patients (31.6%) in the BMS-275291 group (lower limit of one-sided 95% CI: 19.3) and in 11 patients (29.7%) in the placebo treatment arm. The incidence of rash was higher in patients receiving BMS 275291 (28.9% versus 18.9%). An objective response rate of 21.9% was observed in the BMS-275291 treatment arm and 36.4% in the placebo arm. CONCLUSION: BMS-275291 plus paclitaxel/carboplatin was well tolerated and active in advanced non-small cell lung cancer. Treatment with BMS-275291 was not limited by drug-related arthrotoxicity and tumor response was as expected. As planned, patient accrual continued to further investigate the effect of BMS-275291 on overall and progression-free survival in a phase III setting. PMID- 15541823 TI - Results of a Phase II study of carboplatin plus gemcitabine in patients with untreated extensive small cell lung cancer. AB - PURPOSE: To determine the response rate (RR) and survival produced by carboplatin + gemcitabine therapy in patients with untreated extensive small cell lung cancer (ESCLC). PATIENTS AND METHODS: Treatment consisted of carboplatin (AUC = 5) on day 1 and gemcitabine (1100 mg/m(2)) on days 1 and 8 of each 21-day cycle for 4 planned cycles (additional cycles allowed as per treating physician). ECOG performance status 0/1/2 was 29, 58, and 13%. Median age was 66.5 years (range: 41.3-83.1), 94% were white, and 50.7% were female. RESULTS: Between August 2000 and February 2002, 69 patients with ESCLC were enrolled. All 69 patients were included in the safety analysis, and 66 patients were evaluable for response. There were 2 CR (3.0%), 26 PR (39.5%), 23 SD (34.8%), and 15 PD (22.7%) resulting in a RR of 42.5%. The median survival was 9.2 months (range: <1-22.6), and the estimated 1-year survival was 33%. The median TTP was 3.9 months (range: <1 12.8), and the estimated 6-month progression free survival was 24%. The median duration of response was 3.8 months (range: 1.0-9.9). Out of 69 patients, 29, 3, and 16 received 4, 5, and 6 cycles of therapy, respectively. The major Grade 3, 4 toxicities included neutropenia (39.1%), thrombocytopenia (31.9%), anemia (13.0%), and fatigue (4.3%). CONCLUSION: This regimen resulted in survival data that was similar to other regimens for ESCLC and treatment appeared to be well tolerated. Gemcitabine in combination with carboplatin or other active drugs in ESCLC may be worth further investigation. PMID- 15541824 TI - Combination chemotherapy of gemcitabine and vinorelbine in patients with advanced non-small cell lung cancer. PMID- 15541825 TI - Is postoperative radiotherapy for radically resected NSCLC detrimental and therefore unnecessary? PMID- 15541827 TI - Carcinoma in situ from the view of complete resection. AB - World Health Organisation (WHO) defined three types of preinvasive epithelial lesions, one of which is preinvasive bronchial squamous lesions consisting of dysplasia and carcinoma in situ (CIS). It is not clear whether or not CIS at the bronchial resection margin is to be considered as incomplete resection in the literature. Follow-up data of such patients using autofluorescence bronchoscopy proved that CIS regresses without further treatment in significant number of patients. It is therefore reasonable to accept any reported CIS lesion on frozen section examination as complete clearance of the tumor and thus further resection may not be warranted. PMID- 15541828 TI - Epithelial-directed drug delivery: influence of formulation and delivery devices. AB - Technology to deliver drugs reliable throughout the entire respiratory tract has greatly improved in part because of asthma- and infectious disease-related applications. Recent improvements with spiral CT technology applied to detect early lung cancer in high risk populations have also enhanced the prospect of finding locally confined primary lung cancers. In this setting, the need to safely and economically manage the local regional phase of early lung cancer is assuming great strategic importance. At the same time the growing knowledge regarding the molecular and biochemical events driving the progression of lung cancer is allowing the development of targeted drug that may be useful in arresting lung cancer progression. The questions considered in this forum is whether improvements in these areas are sufficiently mature to allow application of local regional drug delivery with targeted drug agents to improve the management of early lung cancer. PMID- 15541829 TI - Impaired glucose tolerance-a new risk factor? PMID- 15541830 TI - The interventionalist's dilemma: innocent intimal hyperplasia or in-stent restenosis? PMID- 15541831 TI - Role of BNP in patients with severe asymptomatic aortic stenosis. PMID- 15541832 TI - The data monitoring experience in the MOXCON trial. AB - AIMS: This article describes a challenging data monitoring experience that occurred in a major international randomized placebo-controlled trial in patients with heart failure, in which the accumulating interim data showed an excess of deaths on the active treatment. METHODS AND RESULTS: The MOXonidine CONgestive Heart Failure trial was a randomized comparison of moxonidine, a central sympathetic inhibitor, with placebo. It was planned to recruit 4500 patients with heart failure. The primary endpoint was all-cause mortality, and average follow up was anticipated to be around 2.5 years until 724 deaths occurred. The trial Data Monitoring Board (DMB) was to conduct safety monitoring reviews of interim data at least every six months, and make their recommendations to the Executive Committee. Within a few months of the study starting, the Data Monitoring Board (DMB) observed an emerging trend of an excess of deaths on moxonidine compared with placebo. This article describes the sequence of events that unfolded: several DMB meetings to evaluate the accumulating evidence, a DMB recommendation to stop the trial, consequent dialogue with the Executive Committee and sponsor leading to a final decision to stop the trial. Ten months after the first patient was randomized, the study was stopped based on 46 versus 25 deaths in 990 moxonidine and 943 placebo patients, respectively, P=0.01. The final published evidence had 54 versus 32 deaths, P=0.012. CONCLUSIONS: This study illustrates the problems faced by a DMB, and subsequently the trial Executive Committee and sponsor, in deciding how to act in the face of an emerging (and agonizing) negative trend for mortality in a major international trial. PMID- 15541833 TI - Regulation of myocardial connexins during hypertrophic remodelling. AB - Cardiac hypertrophic remodelling, initiated by signalling cascades in response to increased workload, injury or intrinsic disease, is initially adaptive. However, prolonged hypertrophy as a consequence of pathological stress leads to maladaptive changes that increase the risk for fatal ventricular arrhythmias. One of these changes is the remodelling of myocardial gap junctions, which provide for electrical coupling of adjacent cardiomyocytes. Myocardial gap junctions are composed of three connexin isotypes, connexin40 (Cx40), -43 (Cx43), and -45 (Cx45) and each display a characteristic developmental and regional expression pattern. Alterations in the distribution and expression of Cx43, the predominant isoform in the adult ventricles, has been the main focus of examination in humans, experimental animal models and cultured cardiomyocytes in response to hypertrophy. The molecular mechanisms and signalling pathways underlying these changes have been studied less thoroughly. In this review we summarize what is known about the remodelling of myocardial gap junctions during hypertrophy, the putative underlying mechanisms and functional consequences thereof. PMID- 15541834 TI - Newly detected abnormal glucose tolerance: an important predictor of long-term outcome after myocardial infarction. AB - AIMS: Recent data revealed that patients with myocardial infarction (MI) have a high prevalence of previously unknown diabetes mellitus (DM) and impaired glucose tolerance (IGT). The added prognostic importance of this finding has not been prospectively explored. To investigate whether a newly detected abnormal glucose tolerance (IGT or DM) assessed early after an MI, is related to long-term prognosis. METHODS AND RESULTS: Patients (n=168; age 63.5+/-9.3 years) with MI, no previous DM and admission blood glucose <11.0 mmol/l were followed for major cardiovascular events defined as the composite of cardiovascular death, non-fatal MI, non-fatal stroke or severe heart failure (HF). According to an oral glucose tolerance test (OGTT) before hospital discharge, 55 patients had normal and 113 abnormal glucose tolerance (GT). During the follow-up of median 34 months there were eight cardiovascular deaths, 15 patients had a recurrent MI, six had a stroke and ten severe HF. All patients who died from cardiovascular causes had abnormal GT. The composite cardiovascular event occurred in 31 (18%) patients. The probability of remaining free from cardiovascular events was significantly higher in patients with normal than abnormal GT (p=0.002). Together with previous MI, abnormal GT was the strongest predictor of future cardiovascular events (hazard ratio 4.18; CI 1.26-13.84; p=0.019). CONCLUSIONS: Abnormal glucose tolerance is a strong risk factor for future cardiovascular events after myocardial infarction. Since it is common and possible to detect even during the hospital phase it may be a target for novel secondary preventive efforts. PMID- 15541835 TI - Association of glomerular filtration rate on presentation with subsequent mortality in non-ST-segment elevation acute coronary syndrome; observations in 13,307 patients in five TIMI trials. AB - AIMS: To determine the association of glomerular filtration rate (GFR) with clinical outcomes in the setting of non-ST-segment elevation acute coronary syndromes (NSTE-ACS). METHODS AND RESULTS: Data were pooled from five NSTE-ACS TIMI trials (TIMI 11A and B, TIMI 12, OPUS-TIMI 16 and TACTICS-TIMI 18) and were available in 13 307 patients. GFR was assessed as a continuous and a categorical variable (normal: > or = 90 mL/min/1.73 m2, n=4952; mildly decreased: 60-89 mL/min/1.73 m2, n=6262; and moderately to severely decreased GFR: <60 mL/min/1.73 m2, n=2093). There was an independent association between decreasing GFR and mortality at 30 days (OR 1.19, 95% CI 1.12-1.27, p<0.001) and at 6 months (OR 1.16, 95% CI 1.11-1.22, p<0.001). The combination of TIMI risk score (TRS) and decreasing GFR provided further mortality risk stratification with highest 30-day and 6-month mortality rates among patients with the lowest GFR who also had a TRS > or = 5 (9.1% and 15.4%, respectively). Decreasing GFR was also independently associated with stroke and recurrent ischaemia at 30-days as well as with major bleeding (p<0.001). CONCLUSION: In the setting of NSTE-ACS, impaired GFR is associated with higher mortality as well as higher rates of thrombotic and major bleeding events, independent of TRS. PMID- 15541836 TI - Troponin is more useful than creatine kinase in predicting one-year mortality among acute coronary syndrome patients. AB - AIMS: To compare the long-term prognostic value of troponins (Tn) vs. conventional cardiac biomarker creatine kinase (CK) and CK-MB across the spectrum of acute coronary syndromes (ACS). METHODS AND RESULTS: In the prospective, observational Canadian ACS Registry, 4627 patients with ACS were enrolled from 51 centres. The CK, CK-MB, Tn samples were analysed in each hospital clinical laboratory and the results related to the reference levels of the individual laboratories. The study cohort comprised 3138 (67.8%) patients who had both CK (or CK-MB) and Tn measurements during the first 24 h of hospitalisation. Vital status at one-year was determined by standardized telephone interview. 61.2% and 59.0% of patients had abnormal Tn and CK (or CK-MB) levels, respectively. Vital status at one-year was ascertained for 2950 patients (6% lost to follow-up). Among patients with normal CK (or CK-MB) levels, elevated Tn was associated with increased one-year mortality (odds ratio [OR] 2.06; 95% CI 1.37-3.11; P=0.001). Similarly, among patients with abnormal CK (or CK-MB) levels, abnormal Tn predicted higher one-year mortality (OR 1.83; 95% CI 1.14-2.93; P=0.01). In contrast, abnormal CK (or CK-MB) was not predictive of mortality after stratification by Tn status. In multivariable analysis controlling for other known prognosticators including creatinine, abnormal Tn (adjusted OR 1.78; 95% CI 1.30-2.44; P<0.001) but not CK/CK-MB was independently associated with increased one-year mortality. CONCLUSIONS: Elevated Tn was independently associated with worse outcome at one-year, while CK or CK-MB status did not provide incremental prognostic information. Our findings support the use of Tn in the risk stratification of unselected ACS patients. PMID- 15541837 TI - Mortality following non-ST elevation acute coronary syndrome: 4 years follow-up of the PRAIS UK Registry (Prospective Registry of Acute Ischaemic Syndromes in the UK). AB - AIM: To present information on long-term prognosis and risk factors following an admission with non-ST elevation acute coronary syndrome. METHODS: A cohort of 653 patients was followed for mortality and causes of death using data from the UK Office of National Statistics (ONS). Cox proportional hazards model was used to identify the prognostic factors. RESULTS: Overall survival at a maximum follow-up of 45 months was 77.8% (95% CI 74.1-81.1%). Seventy-three per cent of the deaths were clearly due to a cardiovascular cause. Age, male gender, heart failure, ST depression or bundle branch block were all associated with higher short- and long term risk. Taking aspirin or having a revascularization procedure, over the period of six months following initial hospitalisation were both associated with a lower long-term risk. CONCLUSION: Non-ST elevation acute coronary syndromes carry a high risk of death over a 4-year period. Conventional risk factors can predict both short- and long-term risk. More invasive management and the use of evidence-based therapies appear to be associated with a lower risk. PMID- 15541838 TI - Preconditioning during coronary angioplasty: no influence of collateral perfusion or the size of the area at risk. AB - AIMS: It is unclear whether the protection observed in human heart following repetition of brief episodes of ischaemia is due to opening of coronary collaterals or to ischaemic preconditioning. We investigated whether the improvement in ST segment change following repeated episodes of brief ischaemia during coronary angioplasty is due to preconditioning when the size of the area at risk and the collateral flow are taken into account. METHODS AND RESULTS: Thirty-six patients underwent percutaneous transluminal coronary angioplasty. Intracoronary ST segment changes were measured throughout the procedure and used as an endpoint. The size of the area at risk and the collateral perfusion within the ischaemic bed were measured using single photon emission computerized tomography (SPECT). Mean ST segment shift observed in all patients significantly decreased from 11.0+/-2.6 mm during the first balloon inflation to 8.5+/-2.3 mm during the second inflation. This protective effect occurred in the absence of any change in the size of the area at risk (mean: 46+/-5% of LV) and of the collateral perfusion to the ischaemic zone (mean: 23+/-4% of flow in the non ischaemic zone). CONCLUSION: These results suggest that ischaemic preconditioning does occur during repeated brief coronary artery occlusion in the human heart. PMID- 15541839 TI - Soft plaque detected on intravascular ultrasound is the strongest predictor of in stent restenosis: an intravascular ultrasound study. AB - AIMS: Although various predictors of in-stent restenosis (ISR) have been reported, the subject of parameters relating to ISR on intravascular ultrasound (IVUS) still leaves room for discussion. The aim of this study was to clarify the strongest predictors of ISR using IVUS. METHODS AND RESULTS: Ninety-two native coronary lesions undergoing single bare-metallic stent implantation were investigated retrospectively. We classified them into the ISR (n=46) and non-ISR (n=46) groups using quantitative coronary angiography. On serial IVUS studies, plaque morphology, and areas and volumes of each component in vessel were evaluated. Among all parameters, diabetes mellitus and soft plaque appearing hypoechoic on IVUS were associated with ISR. By multivariate analysis, soft plaque was the only independent predictor of ISR (p=0.0057). Compared with non soft plaque, soft plaque had a larger plaque reduction rate (-7.1% vs. -1.6%, p=0.0613) and smaller percent plaque volume (53.0% vs. 55.5%, p=0.0273) after stenting. Conversely, soft plaque had a larger neointimal area (4.39 vs. 3.33 mm2, p=0.0437) and percent plaque area (80.5% vs. 75.1%, p=0.0503) at follow-up. CONCLUSION: Soft plaque detected on IVUS was the strongest predictor of ISR. Soft plaque was compressed more easily by stenting, however, causing more proliferation of neointima subsequently and resulted in a worse prognosis. PMID- 15541840 TI - Alpha-adrenergic receptor blockade and hyperaemic response in patients with intermediate coronary stenoses. AB - BACKGROUND: Maximal hyperaemia is paramount in the diagnosis of patients with coronary artery disease. However in these patients, enhanced alpha-adrenergic microvascular vasoconstriction may preclude adenosine to induce maximal hyperaemia. AIM: To assess the presence and the clinical relevance of residual microvascular resistance after administration of adenosine. METHODS AND RESULTS: Fractional flow reserve (FFR, calculated by coronary pressure measurements during adenosine-induced hyperaemia) was assessed in 85 patients with an intermediate coronary stenosis (mean diameter stenosis of 50+/-1%) and normal left ventricular function which were divided into the following three groups: (a) 33 patients before and after IC bolus of phentolamine, an alpha1-, alpha2-adrenergic blocker; (b) 32 patients before and after IC bolus of urapidil, a selective alpha1 adrenergic blocker; (c) 20 patients before and after IC bolus of saline. Since minimal luminal diameter remained unchanged before and after phentolamine (1.46+/ 0.06 vs. 1.47+/-0.06 mm, ns), urapidil (1.46+/-0.06 vs. 1.39+/-0.08, ns), and saline (1.56+/-0.08 vs. 1.55+/-0.08, ns), changes in FFR reflects changes in microvascular resistance. Overall, phentolamine and urapidil induced a slight but significant decrease in FFR (phentolamine: 0.79+/-0.02 vs. 0.77+/-0.02, p<0.05; urapidil: 0.78+/-0.02 vs. 0.75+/-0.02, p<0.05). However, only 6 patients showed a change in FFR from > or = 0.75 to <0.75 and no patients showed a change in FFR from > or = 0.80 to <0.75 that could have influenced clinical decision making. Saline did not induce any change in FFR. Phentolamine and urapidil induced only transient and negligible haemodynamic changes in heart rate and blood pressure. CONCLUSIONS: The administration of alpha-adrenergic blockers in addition to adenosine unmasks a small, yet clinically irrelevant, degree of residual microvascular tone. The consequential changes in FFR values do not significantly affect clinical decision making. PMID- 15541841 TI - Utility of the fractional flow reserve in the evaluation of angiographically moderate in-stent restenosis. AB - AIMS: The evaluation of in-stent restenosis (ISR) is usually based on angiographic quantification. This evaluation is sometimes difficult and it has not an accurate clinical correlation. Fractional flow reserve (FFR) measured by intracoronary pressure wire has demonstrated its value in determining the functional repercussion of coronary stenosis. The aim of this study was to evaluate the relation between quantitative angiography-FFR in borderline in-stent restenotic lesions and the accuracy of FFR in deciding the treatment of ISR. METHODS AND RESULTS: Quantitative angiographic values of 65 lesions in 62 patients with angiographically moderate ISR are compared with the FFR value obtained by pressure wire. An FFR value<0.75 was considered significant. Patients with non-revascularized ISR (FFR > or = 0.75) were clinically followed during a year. An FFR value > or = 0.75 was obtained in 41 lesions (63%), 21 of them with stenosis > or = 50%. The co-efficient of correlation between parameters of quantitative angiography and FFR value was <0.5. No events related to the non treated lesions were observed. CONCLUSIONS: A poor correlation between angiographic quantification and FFR of moderate ISR was found. Conservative management of moderate 40-70% in-stent restenotic lesions with FFR value > or = 0.75 is safe avoiding unnecessary revascularizations based solely on the angiography. PMID- 15541842 TI - Predictors of outcome in patients with severe aortic stenosis and normal left ventricular function: role of B-type natriuretic peptide. AB - AIMS: B-type Natriuretic Peptide (BNP) is activated in patients with severe, symptomatic aortic stenosis (AS), but the prognostic value of BNP in this setting has not been extensively studied. This study aimed to assess the prognostic value of the BNP level in symptomatic and asymptomatic patients with severe AS. METHODS: Seventy consecutive patients referred to our echocardiography laboratory for severe AS with preserved left ventricular function were prospectively enrolled (40 men, median age 74 years [62-82]; aortic valve area 0.7 cm2 [0.6 0.8]; transaortic gradient 48 mmHg [38-60], and left ventricular fractional shortening 38% [32-43]). C-terminal BNP serum level at enrollment was evaluated against baseline functional and echocardiographic parameters as well as clinical outcome. RESULTS: BNP level was elevated in the presence of symptoms and increased with NYHA functional class. BNP serum level >66 pg/ml detected symptomatic patients with a sensitivity, specificity and accuracy of 84%, 82% and 84%, respectively. In symptomatic and asymptomatic patients, BNP level was a strong independent predictor for cardiovascular death by multivariable analysis adjusted to age and NYHA functional class. CONCLUSIONS: BNP serum level allows to differentiate symptomatic from asymptomatic patients with severe AS. BNP is an independent predictor of outcome in these patients and may be helpful for risk stratification. PMID- 15541843 TI - Guidelines on management (diagnosis and treatment) of syncope-update 2004. Executive Summary. PMID- 15541845 TI - Psychological interactions with infertility among women. AB - Despite the fact that various studies have demonstrated the importance of the mind-body connection and fertility, the psychosocial aspects of infertility have not been adequately addressed. Fertility treatments, ranging from medical monitoring, to hormonal remedies and in vitro fertilization (IVF), are both a physical and emotional burden on women and their partners. Psychological factors such as depression, state-anxiety, and stress-induced changes in heart rate and cortisol are predictive of a decreased probability of achieving a viable pregnancy. A couple that is trying to conceive will undoubtedly experience feelings of frustration and disappointment if a pregnancy is not easily achieved. However, if the difficulties progress and the man and or woman are labelled as having fertility problems, then this may result in a severe insult to self esteem, body image, and self-assessed masculinity or femininity. Three types of relationships have been hypothesized between psychological factors and infertility. These include: (1) psychological factors are risk factors of subsequent infertility; (2) the experience of the diagnosis and treatment of infertility causes subsequent psychological distress; (3) a reciprocal relationship exists between psychological factors and infertility. The evidence for these three relationships is reviewed and an alternative approach to the treatment of infertility including stress evaluation that precedes or is concurrent to fertility treatment is suggested. PMID- 15541846 TI - Endometrial adenocarcinoma in young patients: evaluation and fertility-preserving treatment. AB - Carcinoma of the endometrium is the most common female pelvic malignancy in the US. Although it is primarily a disease of the postmenopausal female, 25% of patients are premenopausal, with 3-5% in women 40 years old or younger. The younger group of women with endometrial carcinoma are frequently nulligravid with a history of infertility, and a strong desire to preserve fertility. This may pose a therapeutic dilemma for both patients and treating physicians. Medical treatment for young patients with grade 1 endometrial carcinoma who wish to preserve fertility is a reasonable and appealing option. A comprehensive evaluation prior to counseling the patient should include. A complete history and physical examination. A formal D&C with review of histology with an experienced gyn-onc pathologist. Evaluation of the pelvic and abdomen preferably with contrast-enhanced MRI or transvaginal ultrasound. In patients found to have a clinical stage I grade 1 tumor and who want to preserve fertility, thorough counseling including risks and benefits, and explanation that the data is partial and incomplete due to the lack of appropriate controlled studies is mandatory. In patients considered for medical treatment, a high dose progestin regimen should be started with endometrial sampling every 3 months until complete regression of the tumor is documented. For patients willing to conceive at this stage, treatment options should be discussed. In women who do not want pregnancy at this stage, a maintenance treatment with oral contraceptive agent or Depo-provera (medroxyprogesterone acetate 150 mg i.m. q 12 weeks) should be recommended, with periodic ultrasound evaluation of the endometrium. When the patient finishes her fertility plans, the option of hysterectomy should be considered. Although most responses are long standing, there is a small risk of progression during or after cessation of progestin therapy. PMID- 15541847 TI - Incidence of toxoplasmosis in 5532 pregnant women in Crete, Greece: management of 185 cases at risk. AB - OBJECTIVES: To study the incidence of toxoplasmosis in pregnant women in Crete and to test a designed protocol for handling those at risk of delivering congenitally infected infants. STUDY DESIGN: Pregnant women were screened serologically over a period of 5 years. Cases with suspected acute toxoplasmosis were treated, peripheral blood (PB), and amniotic fluid (AF) tested by polymerase chain reaction (PCR) and culture, and fetuses monitored by ultrasonography. The absence of congenital infection in infants was confirmed by serology and clinical evaluation. RESULTS: Of the 5532 pregnant women followed, 70.57% remained seronegative, 29.45% were seropositive, and there was direct evidence of seroconversion in six cases. Acute toxoplasmosis was suspected in 185 cases, maternal parasitemia was detected in five cases and positive amniotic fluid in one case. Congenital infection was excluded in all infants followed, based on the absence of ultrasound findings in utero, lack of clinical symptoms at birth, negative Western blotting (WB) at birth and 3 months later, and descending serology for a year. CONCLUSION: Overall, 29.45% of the pregnant women followed were seropositive, 3.3% with suspected acute toxoplasmosis, and in 0.02% cases there was evidence of maternofetal transmission. The protocol tested allowed differentiation between acute and latent toxoplasmosis, safe management of the cases at risk and assisted in avoidance of unwarranted pregnancy terminations. PMID- 15541848 TI - Adverse pregnancy outcomes are associated with multiple maternal thrombophilic factors. AB - OBJECTIVE: To determine to what extent adverse pregnancy outcomes are associated with thrombophilia. STUDY DESIGN: We studied 31 women who had HELLP syndrome, placental abruption, fetal growth restriction or unexplained stillbirth (study group), matched with 12 controls. All women were tested for: Factor V, Prothrombin, methylenetetrahydrofolate reductase gene (MTHFR) mutations; for Protein C, S and Antithrombin III deficiency; for lupus anticoagulant. Correlation with 24h BP monitoring and uterine Doppler velocimetry indexes at 22 24 weeks' gestation was performed. RESULTS: Women with multiple thrombophilic factors had a significant lower birth weight (1568.33+/-146.8 g versus 2546.45+/ 438 g), higher 24 h mean diastolic blood pressure at second trimester (76.3+/ 12.5 mmHg versus 65.2+/-7.8 mmHg) and higher RI of uterine arteries (0.69+/-0.05 versus 0.50+/-0.15) than women with single thrombophilic factor. CONCLUSION: Multiple thrombophilic factors carry a major additional risk for adverse maternal and fetal outcomes and correlate well with placental maladaptation as indicated by uterine Doppler velocimetry and 24h BP monitoring. PMID- 15541849 TI - Symmetrical fetal growth retardation after gestational cocaine exposure in the rat. AB - Cocaine use during pregnancy results in an increase in different maternal and perinatal complications. The fetal effects of cocaine could be mainly related to the disturbances in the brain development, microcephaly being the most common brain abnormality. The aim of this study was to analyze maternal outcome and fetal somatic effects of cocaine and to evaluate the hypothesis that maternal cocaine exposure would specifically impair fetal global brain development. Fifty four timed-pregnant female Sprague-Dawley rats were daily injected with 15 or 40 mg/kg per day from gestational day (GD) 1 or 8 and sacrificed at gestational day 20. By analyzing different maternal and fetal outcomes, it could be suggested that the cocaine exposure in pregnant rats decreased maternal weight gain without significant maternal mortality, did not affect the mean number of fetuses by litter, although notably increased stillbirths, reduced fetal birth weight, and reduced the fetal central nervous system weight. Present results are globally in agreement with the literature and underline a possible selective effect of cocaine on the fetal CNS resulting in symmetrical intrauterine fetal growth retardation in contrast to the asymmetrical retardation of undernutrition. PMID- 15541850 TI - Efficacy and safety of a new, chromatographically purified rhesus (D) immunoglobulin. AB - OBJECTIVES: To study the efficacy, safety, and tolerability of the 300 microg dose of a new chromatographically produced rhesus immunoglobulin (Rhophylac 300) for ante- and postnatal rhesus prophylaxis. DESIGN: In an open-label multi-centre study, rhesus D (RhD)-negative women were randomly allocated to receive Rhophylac 300 either intravenously or intramuscularly at the 28th week of gestation and within 72 h after delivery of an RhD-positive child. Serum samples were obtained prior to the antenatal dose and 6-11.5 months after delivery of an RhD-positive child and tested by the indirect antiglobulin test and papain test for anti-D. Safety parameters were assessed in all women who were treated with the study drug. RESULTS: Four hundred and thirty two women received the study drug antenatally. No differences were detected in efficacy or tolerability between intravenous and intramuscular administration. Of the 261 women who delivered an RhD-positive child and received rhesus prophylaxis according to the protocol, 248 women returned for follow-up investigations. None of them had detectable anti-D at their last visit. There were no serious adverse events, no cases of infectious disease transmission nor clinically relevant changes in laboratory safety values and vital signs attributable to the study drug. CONCLUSIONS: The results suggest that Rhophylac 300 given intravenously or intramuscularly is safe and efficacious in preventing rhesus (D) immunisation. PMID- 15541851 TI - Predicting pregnancy-induced hypertension with dynamic hemodynamics. AB - AIM: To study dynamic changes in the hemodynamic parameters in pregnant women during different weeks of gestation and to comprehend changes to the cardiovascular system in normal pregnant women and in patients with pregnancy induced hypertension (PIH), with the object of finding predictors of pregnancy induced hypertension. METHODS: Radialis artery pulse waves of 132 pregnant women were examined from 10 weeks of gestation onward, and the subjects' hemodynamics were examined every 4 weeks. RESULTS: In the women with PIH hemodynamic changes occurred earlier than the onset of disease and even presented in early pregnancy; there might be another mode, and there were factors disturbing the blood pressure modulating mechanism. CONCLUSION: The prediction of PIH from the dynamic changes in hemodynamic parameters has significant implications for clinical practice. PMID- 15541852 TI - Predictive factors of early cessation of breastfeeding. A prospective study in a university hospital. AB - OBJECTIVE: To emphasize the factors involved in early cessation of breastfeeding among patients who have given birth in a high-tech maternity hospital, both during the stay on the maternity ward and before their infants reach the age of 4 months. STUDY DESIGN: Prospective study involving all patients who gave birth during the first 2 months of 2002 in the maternity department of Tours University Hospital, France. Exclusion criteria were preterm delivery before 35 weeks of pregnancy, twin birth and any condition in the newborns that required intensive or long-term hospitalization. The methodology suggested by the World Health Organization in the report "Indicators for Assessing Breastfeeding Practices" was strictly followed, and the breastfeeding status was assessed on the day of discharge from hospital and then in personal telephone conversations with each patient held 1, 2, 3, and 4 months after the date of delivery. The data were analyzed with the Chi-square test or Fisher's Exact Test for the dichotomous variables, and the populations were compared for the continuous variables with the Student test. RESULTS: We studied 115 breastfeeding patients (=57% of the patients who gave birth in the study period). At the time of discharge from hospital, 95% of the 115 were still breastfeeding; the corresponding proportions 1, 2, 3, and 4 months after the birth were 78, 66, 44, and 17%. For patients who were breastfeeding exclusively, the corresponding proportions were: 95, 69, 47, 17, and 6% at discharge from hospital and 1, 2, 3, and 4 months after the birth. We found differences between multiparous and primiparous women only in the timing of the first breastfeeding session and the use of formula supplement while on the maternity ward. There were no differences between the group whose babies were born by cesarean section and those who had vaginal deliveries. The only factor found to be linked with early cessation of breastfeeding during the hospital stay was the lack of previous experience in a multipara who decided to breastfeed for the first time (OR=35.33; 95% CI=2.4531 < OR < 508.925). CONCLUSION: Among the women discharged from the maternity ward with a healthy baby, for a multipara the lack of any previous experience of breastfeeding with earlier babies is the greatest risk factor in early definitive cessation of breastfeeding. Targeting such women would make it possible to provide more support for them when they start breastfeeding and thus hopefully to avoid early cessation. PMID- 15541853 TI - Comparison of fetal heart rate baseline estimation by SisPorto 2.01 and a consensus of clinicians. AB - OBJECTIVE: Fetal heart rate (FHR) baseline estimation is considered one of the most difficult aspects of computer analysis of cardiotocographs, and a pre requisite for the remaining evaluation. This study aimed at comparing FHR baselines assigned by SisPorto 2.01 with those estimated by a consensus of three clinicians. STUDY DESIGN: A prospective evaluation of 300 consecutively acquired FHR tracings was undertaken, 150 from antepartum high-risk pregnancies and 150 from unselected intrapartum cases. Tracings were presented to three experienced clinicians for individual FHR baseline estimation and were processed by the SisPorto 2.01 program. Clinicians were asked to follow a previously described baseline definition, and cases where differences in estimation exceeded 3 bpm were re-evaluated at a consensus meeting. In the remaining cases, the average of the three estimations was considered the consensus value. Agreement between SisPorto 2.01 and the consensus of clinicians was evaluated using the kappa statistic, the proportions of agreement, the intra-class correlation coefficient, and the limits of agreement. RESULTS: Differences < or =3 bpm occurred in 89% of antepartum and 81% of intrapartum cases. No differences exceeding 10 bpm occurred in the antepartum, while in the intrapartum these were found in 1% of cases. Maximum difference between computer and clinicians was 8 bpm in the antepartum and 15 bpm in the intrapartum. Kappa statistics, proportions of agreement and correlation coefficients in the antepartum were 0.97, 0.97 and 0.98, respectively. In the intrapartum, they were 0.87, 0.89 and 0.95. Limits of agreement were -3.22 and 5.22 in the antepartum (mean difference: 1.0; standard deviation: 2.16) and -6.45 and 7.07 in the intrapartum (mean difference: 0.13; standard deviation: 3.45). CONCLUSION: An excellent agreement was found between SisPorto 2.01 and a consensus of clinicians in estimation of FHR baselines both in the antepartum and in the intrapartum. PMID- 15541854 TI - The effect of placental removal method at cesarean delivery on perioperative hemorrhage: a randomized clinical trial ISRCTN 49779257. AB - OBJECTIVES: To evaluate whether the method of placental removal during cesarean section has an impact on perioperative hemorrhage and maternal infectious morbidity. STUDY DESIGN: Three hundred and two patients admitted for abdominal delivery were recruited in a prospective randomized clinical intention-to-treat trial. Participants were assigned to have their placenta removed either manually or spontaneously. The drop in hematocrit was the primary outcome; postpartum maternal infectious morbidity was also assessed. RESULTS: Two patients were excluded for incomplete data. One-hundred-fifty-one were randomized to the manual removal group and 149 to the spontaneous group. The demographic characteristics of the two populations were similar. The mean drop (%) in the manual removal group was greater than in the spontaneous group (5.57+/-3.86 and 2.65+/-2.67, respectively; P<0.01). the incidence of postpartum infectious morbidity was also significantly greater in the manual group (RR 15.8, 95% CI 2.19-117.5). CONCLUSION: Routine manual removal of placenta at cesarean section significantly increases perioperative blood loss and postpartum maternal infectious morbidity. PMID- 15541855 TI - The protective effect of caffeic acid phenethyl ester on ischemia-reperfusion injury in rat ovary. AB - OBJECTIVE: This experimental study was designed to determine the changes in tissue levels of malondialdehyde, end-product of lipid peroxidation (MDA), reduced glutathione (GSH) and xanthine oxidase (XO) and the effect of caffeic acid (3,4-dihydroxycinnamic acid) phenethyl ester (CAPE) on these metabolite levels after adnexal torsion-detorsion model in rats. METHOD: Forty adult female albino rats were divided into five groups: basal control (n = 8), sham operation (n = 8), torsion-detorsion plus saline (n = 8), torsion-detorsion plus CAPE (n = 8). and only torsion (n = 8). Rats in the sham operation group underwent a surgical procedure similar to the other groups but the adnexa was not torsioned. Rats in the torsion group were killed after 360 degrees clockwise adnexal torsion for 3 h and ovaries were harvested. CAPE was injected intraperitoneally 30 min before detorsion in the CAPE/detorsion group and saline was administered in the saline/detorsion group. After 3 h of adnexal detorsion, the rats in both groups were killed and adnexa were surgically removed. RESULTS: MDA levels and XO activities in torsion-detorsion plus saline group increased significantly when compared to basal control, torsion and sham operation groups (P < 0.001). In the CAPE group, MDA levels and XO activities were lower than those of torsion detorsion plus saline group, and differences between the two groups were statistically significant (P < 0.001). GSH levels in torsion-detorsion plus saline group were decreased significantly when compared to basal control and sham operation groups (P < 0.001). GSH levels in the CAPE group were higher than those of torsion-detorsion plus saline group, and differences between the two groups were statistically significant (P < 0.004). Morphologically, polymorphonuclear leukocytic infiltration and vascular dilatation were obvious in the ischemia reperfusion damaged ovary, a change partially reversed by CAPE. CONCLUSIONS: These results suggest that administration of CAPE has beneficial effects in the prevention of ischemia-reperfusion injury of the ovaries. PMID- 15541856 TI - A flexible protocol for cryopreservation of pronuclear and cleavage stage embryos created by conventional in vitro fertilization and intracytoplasmic sperm injection. AB - OBJECTIVES: To evaluate the hospital's cryopreservation protocol. STUDY DESIGN: A retrospective cohort analysis of 30 conventional IVF and 44 ICSI cycles in an assisted conception unit at a tertiary referral hospital. All supernumerary embryos were cryopreserved at the pronuclear or blastomere stage. The survival, morphology, implantation and pregnancy rates were evaluated. The chi2 test and Fisher's exact test were used to determine the statistical significance. RESULTS: A total of 327 pronuclear and cleavage stage embryos were cryopreserved. The post thaw survival rates of 107 conventional IVF and 220 ICSI embryos were 90.6% and 69.0%, respectively. Of the thawed cleavage stage embryos from 43 IVF and 88 ICSI cycles, 90.6% and 69.3% were intact, respectively. Of the thawed pronuclear stage embryos from 64 IVF and 132 ICSI cycles, 90.6% and 68.9% were intact with cleavage rates of 57.8% and 56%, respectively. In the final 27 conventional IVF and 41 ICSI frozen-thawed embryo transfer cycles, the pregnancy rates were 18.5% and 19.5%, respectively. CONCLUSIONS: The adopted cryopreservation protocol flexibly allows for the selection of cleavage-stage embryos for fresh embryo transfer and the cryopreservation of all supernumerary embryos at the pronuclear or the cleavage stage in a single cycle with satisfactory pregnancy rates. Further validation of this protocol is required. PMID- 15541857 TI - Embryo transfer is equally effective at cleavage stage and blastocyst stage: a randomized prospective study. AB - OBJECTIVE: To compare the implantation and pregnancy rates after cleavage stage embryo transfer (ET) with transfer of blastocyst-stage (days 5-6) embryos. STUDY DESIGN: Prospective randomized trial at an assisted reproduction unit in a university hospital. Women with six or more follicles at the last ultrasound scan before oocyte aspiration were randomized for transfer of a maximum of two embryos after 2-3 days (n = 80) or after 5-6 days (n = 64) of culture. Embryo quality, implantation and pregnancy rates were evaluated. Statistical significance was tested with the Chi-square test and Fisher's exact test. RESULT(S): No significant difference was observed in implantation rates (21.1% versus 20.9%, respectively) and clinical pregnancy rates (36.7% versus 32.5% respectively) after blastocyst and cleavage stage transfers for the two groups. The pregnancy rate among subjects who had at least one good quality embryo transferred was 37.5% per day 2-3 ET and 60% per day 5-6 ET. CONCLUSION(S): The overall implantation and pregnancy rates after embryo transfer at cleavage stage and at blastocyst stage transfer were not statistically different. Women who had at least one good quality blastocyst (n = 25) had a high pregnancy rate (60% per ET). Blastocyst transfer is a good alternative for couples with many good quality embryos on day 2 after insemination. PMID- 15541858 TI - Laparoscopic colposuspension using sutures or prolene meshes: a 3-year follow-up. AB - OBJECTIVE(S): To compare the long-term effectiveness of two different laparoscopic colposuspension procedures. STUDY DESIGN: Sixty women affected by genuine stress incontinence (GSI) were enrolled in a prospective randomized controlled trial (RCT) and treated by transperitoneal laparoscopic colposuspension using nonabsorbable sutures (group A) or prolene mesh fixed with tackers or staplers (group B). In each group the subjective and objective failure rates were evaluated at 12, 24, and 36 months after surgery. For the subjective evaluation patients were asked whether they had experienced urine leakage and any urine loss they reported was recorded on a visual analogue scale (VAS). The objective evaluation was performed by means of a clinical examination and multichannel urodynamic studies. The data were analyzed by the intention-to-treat method. RESULTS: The subjective failure rate was significantly (P < 0.05) lower in group A than in group B at 12 months (3.3% versus 13.3%, respectively), 24 months (20.0% versus 36.7%, respectively), and 36 months (33.3% versus 53.3%, respectively) after surgery. The objective failure rate also differed significantly (P < 0.05) between the two groups after 12 (10.7% versus 25.0% for group A and group B, respectively), 24 (29.6% versus 57.7%, respectively), and 36 (42.3% versus 61.5%, respectively) months of follow-up. CONCLUSION(S): Laparoscopic colposuspension performed with sutures is more effective than laparoscopic colposuspension accomplished with the use of prolene meshes in the long term, and the use of prolene meshes should be avoided in treatment of GSI. PMID- 15541859 TI - Age-related complaints in women and their determinants based on a representative community study. AB - OBJECTIVES: To determine (a) the relationship between physical and somatoform complaints, distress, life satisfaction and ageing in the female community and (b) to identify their psychosocial determinants. METHODS: Two stratified random samples of the German female population (total of 2771 women) were investigated by standardised questionnaires of complaints (MFI-20, GBB), distress (HADS) and life satisfaction (LSQ). RESULTS: When participants were divided into six age groups (18-30, 31-40, 41-50, 51-60, 61-70, >70 years), we found a continuous increase of physical, mental and general fatigue, inactivity and reduced motivation. Age-associated increases were also observed for exhaustion, cardiovascular and musculoskeletal complaints, but not for gastrointestinal complaints. This was accompanied by a reduced satisfaction with health and sexuality and increased depression and anxiety scores. Marked increases of complaints were mostly found in the sixth decade. However, depression already increased in the fourth decade, musculoskeletal complaints and reduced motivation peaked in the fifth decade, whereas mental fatigue did not increase significantly before the seventh decade followed by a rise of anxiety in women over 70 years. Most consistently, complaints were predicted by a combination of negative subjective health, higher age, lack of a partnership, and additional sociodemographic vulnerability (unemployment, low income, residence) and protective (religion) factors. CONCLUSIONS: Community data provide important reference points in evaluating the ageing female. Based on regression analyses, we could also demonstrate the contribution of psychosocial vulnerability and protective factors to the development of age-related symptoms. PMID- 15541860 TI - Evaluation of causes of increased incidence of cervical cancer in Slovenia. AB - OBJECTIVE: In Slovenia the opportunistic cervical cancer (CC) screening programme has been performed within the regular gynaecological practice since 1960. The incidence rate 28.8/10(5) decreased to 16.1/10(5) in 1982, and increased to 23/10(5) in 1996. To explain the increased CC incidence the patients' screening histories, occurrence of symptoms, and details of preventive measures were studied on the basis of medical records and questionnaire. Errors of cytological screening were analysed by reviewing previous smears. MATERIAL: The answers obtained from 324 women, treated for CC in the period 1995-2000, were analysed in relation to age, stage and histology. The mean women's age was 46.76 +/- 13.07 (S.D.) years (range 23-85 years). After therapy, 271 patients were followed-up by the end of January 2004, (mean follow-up time 80.3 +/- 20 S.D. months, range 29.7 110.3 months). Statistical analysis was performed using Chi-square test. RESULTS: In 208 (80%) cases, CC was detected in stage 1, in 43 (17%) in stages 2A to 3B. Squamous-cell carcinoma (192, 74.1%) was followed by adenocarcinoma (61, 23.5%) and (5, 1.9%) other malignancies. The screening interval ranged from 6 months to 4 years. Statistically significant differences existed between clinical and screening variables. CC was diagnosed in higher stages in women who were ignorant of the Pap test. Re-screened smears (n = 126) showed 27.8% of false negative results. CONCLUSIONS: The reasons for high incidence of CC are poor performance of cytological screening, failures in gynecological examinations and diagnostic procedures, and the patients' negligence of attending regular screening. PMID- 15541861 TI - Risk factors for bacterial vaginosis. AB - OBJECTIVE: To analyse risk factors for bacterial vaginosis (BV). STUDY DESIGN: We conducted a case-control study on risk factors for BV, in several first-level gynecological centers in Italy. Cases were women aged 18-70 years with confirmed diagnosis of BV (using test for proline iminopeptidase (PIP) activity). Control were the first patients observed in the same center without any symptom suggesting vaginal infection, and a negative PIP activity test. A total of 476 cases and 450 controls entered the study. RESULTS: The number of sexual partners in the month before interview, but not the number of intercourses, was associated with an increased risk of BV. The risk of BV with vaginal douching or tight jeans/trousers once or more a week was 2.0 (95% CI 1.0-3.9) and 1.5 (95% CI 1.0 2.2), respectively, compared to never users. CONCLUSION: The findings are consistent with previous data and suggest that daily habits play an important role in risk of BV. PMID- 15541862 TI - Longitudinal and cross-sectional studies of HIV-1 RNA and DNA loads in blood and the female genital tract. AB - OBJECTIVE: To examine if correlates of HIV-1 genital shedding in cross-sectional studies can be used to determine the risk of shedding in individual HIV-1 positive women. STUDY DESIGN: Longitudinal samples from blood and cervix were obtained from 18 HIV-1 infected women, and HIV-1 RNA and cell-associated DNA virus, and beta-chemokine levels, were measured. Associations between variables were analyzed at both individual and group level. RESULTS: The variation over time was 2.9-, 2.1-, and 2.3-fold in plasma RNA, PBMC DNA and cervical RNA load, respectively, and reached 6.2-fold in cervical DNA load. Differences were observed between associations in individual- and group-level comparisons, suggesting that a separate reservoir of HIV replication may exist in the genital tract of some women, which is influenced by local environmental factors. CONCLUSIONS: Our study underscores the importance of caution during contact with genital fluids at all stages of infection and disease regardless of treatment and HIV-1 blood loads. PMID- 15541863 TI - Endometriosis with FDG uptake on PET. AB - BACKGROUND: The value of [18F]fluorodeoxyglucose positron emission tomography (FDG-PET) uptake in endometriosis has not yet been extensively reported. CASE REPORT: A 32-year-old woman was examined to find an explanation for right pelvic pain associated with right subcostal pain. A computerised tomography (CT) scan was compatible with a haemangioma or a focus of endometriosis in the liver. Transvaginal sonography and magnetic resonance imaging (MRI) showed a complex ovarian cyst on the left. Blood CA125 levels were elevated. FDG-PET revealed a focus of uptake in the right paravesical area. Laparoscopy showed a left endometrioma associated with diffuse inflammatory pelvic adhesions. After surgery and 3 months GnRH agonist treatment the pain had disappeared and neither MRI nor FDG-PET showed any pelvic abnormality. The patient subsequently presented with dyspareunia and rectal pain resulting from a right uterosacral nodule and a rectal nodule. These were resected laparoscopically. After a 1-year follow-up, the patient is doing well. CONCLUSION: Endometriosis can give rise to false positive results on FDG-PET. However, the FDG uptake in this particular case of endometriosis seems to have been due to inflammation rather than to a cyst. This report highlights the relationship between some of the biological features of endometriosis and some observed in neoplastic lesions. PMID- 15541864 TI - Preeclampsia and HIV infection. PMID- 15541865 TI - Bilateral pelvic ganglioneuroma: clinico-pathological findings: a case report. AB - Ganglioneuroma is a rare benign neurogenic tumor originating from the autonomic nervous system, and is considered the benign counterpart of neuroblastoma, lacking the immature neuroblastic cells. A case of pelvic ganglioneuroma with bilateral pelvic spread is described. PMID- 15541866 TI - Medical treatment of abdominal pregnancy. PMID- 15541867 TI - Emergency pelvic packing to control massive intraoperative bleeding during pelvic posterior exenteration. PMID- 15541868 TI - Unilateral cervico-vaginal atresia with ipsilateral renal agenesis. PMID- 15541869 TI - Traumatic rupture of gravid uterus with fetus lying into abdominal cavity: is easy to diagnose? PMID- 15541871 TI - Hospital visits: the trainee's perspective. PMID- 15541872 TI - Specialist life--Emile Papiernik. PMID- 15541873 TI - Microinjection of urocortin 2 into the dorsal raphe nucleus activates serotonergic neurons and increases extracellular serotonin in the basolateral amygdala. AB - The intra dorsal raphe nucleus (DRN) administration of corticotropin releasing hormone (CRF) inhibits serotonergic (5-HT) activity in this structure, an effect blocked by antagonists selective for the type 1 CRF receptor (CRF1). The DRN has a high density of the type 2 receptor (CRF2), and so the present experiments explored the impact of CRF2 activation within the DRN on 5-HT function. The intra DRN administration of the selective CRF2 agonist urocortin 2 (Ucn 2) dose dependently increased 5-HT efflux in the basolateral amygdala, a projection region of the DRN. Intra-DRN Ucn 2 also increased c-fos expression in labeled 5 HT neurons. Both of these effects of Ucn 2 were completely blocked by intra-DRN antisauvagine-30 (ASV-30), a relatively selective CRF2 antagonist. These data suggest that CRF1 and CRF2 activation within the DRN affect 5-HT neurons in opponent fashion. Implications of these results for understanding the behavioral effects of CRF and other CRF-like ligands are discussed. PMID- 15541874 TI - Attenuation of alpha2A-adrenergic receptor expression in neonatal rat brain by RNA interference or antisense oligonucleotide reduced anxiety in adulthood. AB - Brain alpha2-adrenergic receptors (alpha2-ARs) have been implicated in the regulation of anxiety, which is associated with stress. Environmental treatments during neonatal development could modulate the level of brain alpha2-AR expression and alter anxiety in adults, suggesting possible involvement of these receptors in early-life programming of anxiety state. The present study was undertaken to determine whether the reduction of the expression of A subtype of these receptors most abundant in the neonatal brain affects anxiety-related behavior in adulthood. We attenuated the expression of alpha2A-ARs during neonatal life by two different sequence specific approaches, antisense technology and RNA interference. Treatment of rats with the antisense oligodeoxynucleotide or short interfering RNA (siRNA) against alpha2A-ARs on the days 2-4 of their life, produced a marked acute decrease in the levels of both alpha2A-AR mRNA and [3H]RX821002 binding sites in the brainstem into which drugs were injected. The decrease of alpha2A-AR expression in the neonatal brainstem influenced the development of this receptor system in the brain regions as evidenced by the increased number of [3H]RX821002 binding sites in the hypothalamus of adult animals with both neonatal alpha2A-AR knockdown treatments; also in the frontal cortex of antisense-treated, and in the hippocampus of siRNA-treated adult rats. These adult animals also demonstrated a decreased anxiety in the elevated plus maze as evidenced by an increased number of the open arm entries, greater proportion of time spent in the open arms, and more than a two-fold increase in the number of exploratory head dips. The results provide the first evidence that the reduction in the brain expression of a gene encoding for alpha2A-AR during neonatal life led to the long-term neurochemical and behavioral alterations. The data suggests that alterations in the expression of the receptor-specific gene during critical periods of brain development may be involved in early-life programming of anxiety-related behavior. PMID- 15541875 TI - Neuronal cytoskeletal alterations in an experimental model of depression. AB - It has been proposed that depression is associated with hippocampal morphological changes. The apical dendrite atrophy of hippocampal CA3 pyramidal neurons has been described in experimental models of depression. The aim of the present study was to determine which cytoskeletal components are involved in the morphological changes previously described in the hippocampus of depressed animals. The expression of different neuronal cytoskeletal markers was analyzed by immunohistochemistry in rats exposed to a learned helplessness paradigm, an experimental model of depression. Rats were trained with 60 inescapable foot shocks (0.6 mA/15 s) and escape latencies and failures were tested 4 days after training. Animals in which learned helplessness behavior persisted for 21 days were included in the depressed group. No foot shocks were delivered to control rats. Microtubule-associated protein 2 (MAP-2) and light (NFL; 68 kDa), medium (NFM; 160 kDa) and heavy (NFH; 200 kDa) neurofilament subunit immunostainings were analyzed employing morphometric parameters. In the depressed group, NFL immunostaining decreased 55% (P<0.05) and 60% (P<0.001) in CA3 and dentate gyrus, respectively. In the same areas, MAP-2, NFM and NFH immunostainings did not differ between depressed and control animals. Since NFL is present in the core of mature neurofilament, it is proposed that hippocampal depression-associated plastic alterations may be due to changes in the dynamics of the neurofilament assembly. PMID- 15541876 TI - Effects of medial prefrontal cortex and dorsal striatum lesions on retrieval processes in rats. AB - Exposure to training-related cues is known to reactivate associated memory and improves subsequent retention performance under various circumstances. The present studies investigated the neural basis of retrieval cue effects, by studying in two separate experiments, the involvement of the medial prefrontal cortex and of the dorsal striatum. Rats with lesions to the prelimbic-infralimbic cortex (PL-IL), to the anterior dorsal cingulate (ACd), and to the lateral and medial parts of the dorsal striatum (lDS and mDS) were first trained in a brightness discrimination avoidance task. One day later, rats were tested after being placed in the cueing box with either no training-related cue or with additional exposures to the light discriminative stimulus. None of the lesions affected the acquisition performance. During the retention test, control rats cued with the light in the box exhibited significantly better retention performance than those simply placed in the box, confirming our previous results. While mDS lesions did not modify effects of the retrieval cue, lDS as well as both PL-IL and ACd lesions blocked the facilitative effects of the discriminative stimulus. The present data indicate that ACd, PL-IL and lDS are involved in processes promoted by exposure to training cues, the nature of which are reviewed and discussed. This study in conjunction with previous ones suggests that retrieval cues activate several subcircuits mainly based on an amygdalo prefrontal-striatum network. Activation of this network results in an improvement of the expression of the associated conditioned response, and may thus be viewed as increasing the efficacy of the retrieval processes. PMID- 15541877 TI - Behavioural endocrine immune-conditioned response is induced by taste and superantigen pairing. AB - Administration of bacterial superantigen, such as staphylococcal enterotoxin B (SEB), induces in vivo stimulation of T cell proliferation and cytokine production such as interleukin-2 (IL-2). It has been previously reported that SEB administration induces fever, c-Fos expression in the brain, and hypothalamus pituitary-adrenal axis activation, demonstrating that the brain is able to sense and respond to SEB. Previously it had been shown that immune functions can be behaviourally conditioned pairing a novel gustatory stimulus together with an immunomodulatory drug or an antigen. We designed an experimental protocol using Dark Agouti rats in which saccharin taste, as conditioned stimulus, was paired with an i.p. injection of SEB (2 mg/kg), as unconditioned stimulus. Six days later, when conditioned animals were re-exposed to the conditioned stimulus they displayed strong conditioned taste avoidance to the saccharin. More importantly, re-exposure to the conditioned stimulus significantly increased IL-2, interferon gamma and corticosterone plasma levels, in comparison with conditioned animals which had not been re-exposed to saccharin taste. These results demonstrate a behavioural-immune-endocrine conditioned response using a superantigen as unconditioned stimulus. In addition, they illustrate the brain abilities to mimic the unconditioned effects of a superantigen by yet unknown mechanisms. PMID- 15541878 TI - Behavioral characterization in a comprehensive mouse test battery reveals motor and sensory impairments in growth-associated protein-43 null mutant mice. AB - The growth-associated protein (GAP)-43 is a major neuronal protein associated with axonal growth, neuronal plasticity and learning. The observation that only 5 10% of mice with a full GAP-43 gene deletion survive weaning suggests that basic neural functions are disturbed. Here we used a comprehensive test battery to characterise and quantify the motor and sensory function of surviving adult homozygous GAP-43 (-/-) mice as compared with GAP-43 (+/-) and wild-type animals. The test battery was comprised of motor, sensory, and reflex tests producing 25 measures of locomotion, as well as epicritic, auditory, olfactory and visual function. The analysis revealed significant impairments in muscle strength, limb coordination and balance in GAP-43 (-/-) mice. Furthermore, GAP-43 (-/-) animals were hyperactive and showed reduced anxiety as measured by open field and light dark tests. In sensory tests, GAP-43 (-/-) mice were tested for impaired tactile and labyrinthine function. Abnormal reflexes were found in the contact and vibrissa placing responses, and in the crossed extensor reflex. GAP-43 (+/-) animals showed only moderate abnormalities as compared with wild-type animals. We conclude that GAP-43 is necessary for the development and function of a variety of neuronal systems. The results also show that the comprehensive test battery used in the present study represents a sensitive approach to assess the functional integrity of ascending and descending pathways in genetically manipulated mice. PMID- 15541879 TI - Impaired passive avoidance learning in mice lacking central neuronal nicotinic acetylcholine receptors. AB - The nicotinic cholinergic system influences cognition, anxiety, locomotion, and addiction by acting upon nicotinic acetylcholine receptors (nAChRs). To date, there are 12 known neuronal mammalian nAChR subunits leading to a rich pharmacological diversity that is difficult to attribute to specific subunits. We generated alpha7-beta2 nAChR double mutant mice by breeding to investigate the effect of a minimal number of nAChRs in the CNS. These mice have been used to determine the role these receptor subunits play in a variety of behaviors. A battery of behavioral tests was used to determine the effect of the mutation in anxiety, locomotor activity, startle response, pre-pulse inhibition, motor coordination and learning and memory. Mice lacking both the alpha7 and the beta2 nAChR subunits displayed impaired learning and memory performance in a passive avoidance test and showed enhanced motor performance on the rotarod. PMID- 15541880 TI - Altered mitogen-activated protein kinase signaling, tau hyperphosphorylation and mild spatial learning dysfunction in transgenic rats expressing the beta-amyloid peptide intracellularly in hippocampal and cortical neurons. AB - The pathological significance of intracellular Abeta accumulation in vivo is not yet fully understood. To address this, we have studied transgenic rats expressing Alzheimer's-related transgenes that accumulate Abeta intraneuronally in the cerebral and hippocampal cortices but do not develop extracellular amyloid plaques. In these rats, the presence of intraneuronal Abeta is sufficient to provoke up-regulation of the phosphorylated form of extracellular-regulated kinase (ERK) 2 and its enzymatic activity in the hippocampus while no changes were observed in the activity or phosphorylation status of other putative tau kinases such as p38, glycogen synthase kinase 3, and cycline-dependent kinase 5. The increase in active phospho-ERK2 was accompanied by increased levels of tau phosphorylation at S396 and S404 ERK2 sites and a decrease in the phosphorylation of the CREB kinase p90RSK. In a water maze paradigm, male transgenic rats displayed a mild spatial learning deficit relative to control littermates. Our results suggest that in the absence of plaques, intraneuronal accumulation of Abeta peptide correlates with the initial steps in the tau-phosphorylation cascade, alterations in ERK2 signaling and impairment of higher CNS functions in male rats. PMID- 15541881 TI - Increased P27KIP1 protein expression in the dentate gyrus of chronically stressed rats indicates G1 arrest involvement. AB - Various chronic stress paradigms decrease new cell proliferation in the hippocampal dentate gyrus, yet the exact underlying mechanism is still unclear. In the first gap (G1) phase of the cell cycle, both stimulatory and inhibitory signals derived from the extracellular environment converge. Corticosteroids, which increase during stress and are well-known anti-mitotics, cause cells in vitro to arrest in the G1 phase. Following 3 weeks of unpredictable stress, we therefore expected a change in protein expression of various important G1 cell cycle regulators in the adult rat subgranular zone. Using quantitative immunocytochemistry, we show that particularly cyclin-dependent kinase inhibitor p27Kip1 expression is significantly increased. In addition, 3 weeks of recovery after stress normalized the numbers of p27Kip1-expressing cells, consistent with the recovered adult cell proliferation in these animals. P27Kip1-positive cells do not overlap with GFAP-staining and only to a limited extent with Ki-67 expressing cells. Numbers of cyclin E- and cyclin D1-expressing cells did not change after chronic stress. These results indicate that chronic stress causes cycling cells in the adult hippocampus to arrest in G1, thereby providing more mechanistic insight in the stress-induced decrease in cell proliferation. PMID- 15541882 TI - Molecular and functional analysis of hyperpolarisation-activated nucleotide-gated (HCN) channels in the enteric nervous system. AB - Hyperpolarisation-activated non-specific cation currents (Ih currents) are important for the regulation of cell excitability. These currents are carried by channels of the hyperpolarisation-activated nucleotide-gated (HCN) family, of which there are four known subtypes. In the enteric nervous system (ENS), the Ih current is prominent in AH neurons. We investigated the expression and localization of HCN isoforms in the ENS of mice, rats and guinea-pigs. HCN1, HCN2 and HCN4 were expressed in enteric neurons. Immunoreactivity for HCN1 was observed on neuronal cell membranes of Dogiel type II neurons in rat and mouse. HCN2 channel immunoreactivity occurred in the majority of enteric neurons in the guinea-pig, rat and mouse. Immunoreactivity for HCN4 protein was revealed on the cell membranes of many neurons, including Dogiel type II neurons, in the guinea pig. HCN4 was expressed by glial cells in guinea-pig. There was no evidence of HCN3 channel protein in any species with either immunohistochemistry or Western analysis. RT-PCR (polymerase chain reaction) using mouse HCN primers revealed mRNA for all four channels in the longitudinal muscle plus myenteric plexus of mouse distal colon. Sequencing confirmed the identity of the mRNA. Quantitative PCR demonstrated that HCN2 was the most highly expressed HCN channel subtype in the myenteric plexus of mouse distal colon. HCN1 and HCN4 were expressed at lower levels. HCN3 subtype mRNA was 0.2% of HCN2. We used intracellular recording to identify neurons having Ih currents and intracellular dye filling to locate the neurons for the immunohistochemical determination of channel expression. AH neurons with Ih currents were HCN2 and HCN4 channel positive. There was no correlation between the magnitude of the Ih and intensity of channel immunoreactivity. Our results indicate that HCN1, 2 and 4 genes and protein are expressed in the ENS. AH/Dogiel type II neurons, which have a prominent Ih, express HCN2 and 4 in guinea-pig and HCN1 and 2 in mouse and rat. PMID- 15541883 TI - Gamma-secretase subunit composition and distribution in the presenilin wild-type and mutant mouse brain. AB - Studies conducted in cell culture indicate that the gamma-secretase involved in amyloid beta-formation and Notch signaling is a multisubunit aspartic protease. Little is known, however, of the structure, function, or localization of gamma secretase in the adult brain, or possible effects of familial Alzheimer's disease (FAD)-causing mutations on the brain protease. We report here that mouse brain contains a complex composed of gamma-secretase subunits presenilin-1 N-terminal fragment, presenilin-1 C-terminal fragment, Nicastrin, Aph-1a and Pen-2. A homozygous FAD-linked Presenilin-1 knock-in mutation does not alter relative subunit levels. Immunocytochemical localization of gamma-secretase subunits revealed overlapping but distinct regional and subcellular distributions. All subunits are expressed throughout the neuraxis predominantly in neurons, and are present in axons. Their distributions and levels of expression are unaffected by mutant presenilin-1. In a presenilin-1/amyloid precursor protein double knock-in mouse, subunits are associated with plaques, but are expressed at similar levels in amyloid-rich and -poor regions. gamma-Secretase subunits are distributed much more extensively than circumscribed amyloid deposits, suggesting the importance of other factors for localized amyloid deposition. These results indicate a widespread neuronal function for gamma-secretase in the adult brain, and suggest the pathogenic mechanism of FAD-linked mutations does not involve alterations in the composition, expression or brain distribution of the protease. The subcellular localization of gamma-secretase subunits is consistent with a nerve terminal source for amyloid aggregates. PMID- 15541884 TI - Rapid actions of estradiol on cyclic amp response-element binding protein phosphorylation in dorsal root ganglion neurons. AB - Actions of gonadal steroids have not been widely investigated in the peripheral nervous system, although many dorsal root ganglion (DRG) and autonomic pelvic ganglion (PG) neurons express estrogen receptors (ERs). We have studied the effects of 17beta-estradiol exposure on cultured DRG and PG neurons from adult rats. Western blotting analysis of DRG extracts detected phosphorylation of ERK1 and ERK2 (extracellular signal-regulated kinases) that peaked 10 min after exposure to 17beta-estradiol. These extracts contain both neurons and glia; therefore, to determine if this response occurred in DRG neurons, we developed an immunocytochemical method to specifically measure activation in individual neurons. These measurements showed that estradiol increased phosphorylation of CREB (cyclic AMP response-element binding protein), which was consistently blocked by the ERK pathway inhibitor PD98059 but not by the inhibitors of phosphatidylinositol 3-kinase, wortmannin and LY294002. 17beta-Estradiol activation of CREB in DRG neurons was reduced by the ER antagonist, ICI182780. In contrast, in PG neurons estradiol did not affect CREB phosphorylation, highlighting a difference in E2 responses in different populations of peripheral neurons. This study has shown that estrogens can rapidly activate signaling pathways associated with CREB-mediated transcriptional regulation in sensory neurons. As these pathways also mediate many effects of neurotrophic factors, changes in estrogen levels (e.g. during puberty, pregnancy or menopause) could have broad-ranging genomic and non-genomic actions on urogenital pain sensation and reflex pathways. PMID- 15541885 TI - Dual effect of GABA on descending monosynaptic excitatory postsynaptic potential in frog lumbar motoneurons. AB - Monosynaptic excitatory postsynaptic potentials (EPSPs) evoked by stimulating ipsilateral ventrolateral column (VLC) in the thoracic section were recorded in lumbar motoneurons within the isolated spinal cord of the frog Rana ridibunda. Bath application of the selective GABAB receptor agonist (-)-baclofen (0.05 mM) caused a reduction in the peak amplitude of VLC EPSP. Baclofen did not cause any consistent change in the membrane potential or in the EPSP waveform within frog motoneurones. The selective GABA(B) receptor antagonist saclofen (0.1 mM) completely blocked the effect of (-)-baclofen on VLC EPSP. A decrease in VLC EPSP peak amplitude was also observed during GABA (0.5 mM) application. Unlike (-) baclofen, inhibition of VLC EPSP induced by GABA was accompanied by a shortening of the EPSP time course and a reduction in membrane input resistance within lumbar motoneurons. The decrease in VLC EPSP peak amplitude induced by (-) baclofen and GABA was accompanied by an increase in the paired-pulse facilitation. These data provide evidence for a dual pre- and postsynaptic GABAergic inhibition of the VLC monosynaptic EPSP in lumbar motoneurons within the frog spinal cord. PMID- 15541886 TI - Contribution of different calcium channels to long-term potentiation in superior cervical ganglion of the rat. AB - We explored the contribution of different calcium channel types to the long-term potentiation (LTP) of superior cervical ganglion of the rat. Right after a conditioning train of 40 Hz for 5 s, the maximum amplitude of the postsynaptic response (maximum potentiation) increased 5.6+/-0.5-fold. Potentiation decreased to 20% of its initial value within the following 70.0+/-8.0 min (LTP decay time). The contribution of P/Q-, N- and L-type calcium channels to LTP was studied by blocking their activity with synthetic funnel-web spider toxin (10 or 100 microM), omega-conotoxin GVIA (5 microM) or nifedipine (10 microM), respectively. The three blockers reduced the amplitude of the postsynaptic compound action potential before the conditioning train. After the train, all of the toxins reduced the LTP decay time and the integral of the amplitude versus time curve, defined as the LTP extent. In addition, all three blockers increased the maximum potentiation. Our results demonstrate that different calcium channel types contribute to ganglionic LTP. These effects may be by coupling excitation secretion from different types of synaptic vesicles. PMID- 15541887 TI - Developmental plasticity of rat cerebellar cortex after cisplatin injury: inhibitory synapses and differentiating Purkinje neurons. AB - A single injection of cisplatin, a cytostatic agent, (5 microg/g body weight) in 10-day old rats leads later to the reorganization of the cerebellar cortex in lobules VI-VIII of the vermis. Double immunofluorescence reaction for glutamate receptor (GluR)2/3, a ionotropic glutamate receptor that labels postsynaptically Purkinje neurons, and glutamic acid decarboxylase (GAD)65, an isoform of the GABA synthesis enzyme that labels presynaptically inhibitory terminals in the molecular layer, were employed. Less-differentiated Purkinje cells were present in rats treated on postnatal day (PD)11 at the top of lobule VI and in lobules VII-VIII, in comparison with the deep zones of the same lobules and lobule III. The changes were interpreted as due to loss of trophic factors of Purkinje cell growth, e.g. signaling molecules and granule cells. However, we have shown that a remodelling of Purkinje cell dendrites occurred on PD30 (20 days after cisplatin). In fact, despite of the GluR2/3 labeling of the entire Purkinje cell dendrites, the GAD65 immunofluorescent terminals were adjacent to the proximal parts of the dendrite, while they were scarce in the distal dendritic branchlets. The findings were discussed in relation to the changed cytoarchitecture of the cerebellar cortex, which from PD17 to PD30 includes regeneration of the external germinal layer, reorientation of the main dendritic branches and of the Purkinje cell branchlets, and the presence of ectopic cells. PMID- 15541889 TI - Subcortical afferents to the lateral mediodorsal thalamus in cynomolgus monkeys. AB - The mediodorsal (MD) nucleus of the thalamus has long been known to provide the principal source of subcortical input to the primate prefrontal cortex, as well as to other areas of the frontal lobe that are thought to contribute to higher order cognitive functions. In this study, we used injections of retrograde tracers in the lateral portion of the monkey MD to assess the locations of labeled cells in subcortical structures. Three main patterns were identified in the distribution of subcortical connections. We found that the claustrum, superior colliculus and ventral midbrain regions were heavily labeled in the cases with injections in caudoventral MD. In these cases, labeled cells were also found in either the periaqueductal gray or zona incerta, depending on the specific case. In one case with an injection in anterodorsal MD, labeled cells were most numerous in the structures of the ventral midbrain, especially the ventral tegmental area. Finally, the claustrum and superior colliculus contained the largest percentage of labeled subcortical cells in cases with injections in ventrolateral MD. These three patterns of subcortical label corresponded to three equally distinctive trends in the distribution of MD connections with the cortex in these same cases [J Comp Neurol 473 (2004) 107]. Very few labeled cells were found in other areas such as the amygdala, globus pallidus and deep cerebellar nuclei, suggesting that pathways leading from these structures to dorsolateral and dorsomedial frontal cortices are not likely to include the lateral divisions of MD. In concert, these findings show that particular locales within lateral MD receive distinct profiles of subcortical afferents, and project into specific neocortical domains, suggesting that these different sites within lateral MD may participate in functionally distinct circuits of information processing. PMID- 15541888 TI - Effect of hypoxia on the expression and activity of mitogen-activated protein (MAP) kinase-phosphatase-1 (MKP-1) and MKP-3 in neuronal nuclei of newborn piglets: the role of nitric oxide. AB - Mitogen-activated protein kinase-1 (MAPK-1) and MAPK-3 regulate survival and programmed cell death of neurons under stress conditions. The activity of MAPK-1 and MAPK-3 is regulated by dual specificity phosphatases: MKP-1 and MKP-3. In previous studies, we have shown that cerebral hypoxia results in increased activation of MAPK-1 and MAPK-3. Furthermore, we have shown that the hypoxia induced activation of MAPK is nitric oxide (NO)-mediated. The present study tested the hypothesis that hypoxia results in altered expression and activity of MKP-1 and MKP-3 in neuronal nuclei and the administration of 7-nitro-indazole (7 NINA; 1 mg/kg, 60 min prior to hypoxia), a selective nNOS inhibitor, will prevent the hypoxia-induced alteration in the expression and activity of MKP-1 and MKP-3. To test this hypothesis expression and activity of MKP-1 and MKP-3 were determined in neuronal nuclei of normoxic (Nx; n=5), hypoxic (Hx; n=5) and 7-NINA pretreated-hypoxic (7-NINA-Hx; n=5). Hypoxia was achieved by exposing the animals to an FiO2 of 0.07 for 60 min. Cerebral tissue hypoxia was documented biochemically by determining ATP and phosphocreatine levels. Neuronal nuclei were isolated using discontinuous sucrose gradient centrifugation and purified. Nuclear proteins were analyzed by Western blot using specific antibodies for MKP 1 and MKP-3 (Santa Cruz, CA, USA). The protein band density was determined by imaging densitometry and expressed as OD x mm2. The density of MKP-1 was 61.57+/ 5.68, 155.86+/-44.02 and 69.88+/-25.54 in the Nx, Hx and 7-NINA-Hx groups, respectively (P<0.05, ANOVA). Similarly, the density of MKP-3 was 66.46+/-5.88, 172.04+/-33.10 and 116.88+/-14.66 in the Nx, Hx and 7-NINA-Hx groups, respectively (P<0.05, ANOVA). The data show an increased expression of MKP-1 and MKP-3 during hypoxia in neuronal nuclei of newborn piglets and the administration of 7-NINA, an nNOS inhibitor, prevented the hypoxia-induced increased expression of MKP-1 and MKP-3. The activity of MKP-1 (pmol/min) was 176.17+/-16.95 in Nx, 97.56+/-10.64 in Hx and 130+/-14.42 in the 7-NINA-Hx groups, respectively (P<0.05, ANOVA). Similarly the activity of MKP-3 was 104.11+/-12.17 in Nx, 36.29+/-16.88 in Hx and 77.89+/-20.18 in the 7-NINA groups, respectively (P<0.05, ANOVA). The results demonstrate that cerebral hypoxia results in increased expression of MKP-1 and MKP-3 expression that was prevented by the administration of 7-NINA. In contrast, hypoxia resulted in decreased activity of MKP-1 and MKP-3 that was prevented by the administration of a nNOS inhibitor. We conclude that hypoxia-induced decrease in MKP-1 and MKP-3 activity is not due to altered expression but due to NO-mediated modification of the cysteine residue at the active site of these dual specificity phosphatases, a mechanism of their inactivation that leads to activation of MAP kinases. PMID- 15541890 TI - Regeneration influences expression of the Na+, K+-atpase subunit isoforms in the rat peripheral nervous system. AB - Neural injury triggers changes in the expression of a large number of gene families. Particularly interesting are those encoding proteins involved in the generation, propagation or restoration of electric potentials. The expression of the Na+, K+-ATPase subunit isoforms (alpha, beta and gamma) was studied in dorsal root ganglion (DRG) and sciatic nerve of the rat in normal conditions, after axotomy and during regeneration. In normal DRG, alpha1 and alpha2 are expressed in the plasma membrane of all cell types, while there is no detectable signal for alpha3 in most DRG cells. After axotomy, alpha1 and alpha2 expression decreases evenly in all cells, while there is a remarkable onset in alpha3 expression, with a peak about day 3, which gradually disappears throughout regeneration (day 7). beta1 Is restricted to the nuclear envelope and plasma membrane of neurons and satellite cells. Immediately after injury, beta1 shows a homogeneous distribution in the soma of neurons. No beta2 expression was found. Beta3 Specific immunofluorescence appears in all neurons, although it is brightest in the smallest, diminishing progressively after injury until day 3 and, thereafter, increasing in intensity, until it reaches normal levels. FXYD7 is expressed weakly in a few DRG neurons (less than 2%) and Schwann cells. It increases intensely in satellite cells immediately after axotomy, and in all cell types at day 3. Transient switching of members of the Na+, K+-ATPase isoform family elicited by axotomy suggests variations in the sodium pump isozymes with different affinities for Na+, K+ and ATP from those in intact nerve. This adaptation may be important for regeneration. PMID- 15541891 TI - Calcium influx through presynaptic 5-HT3 receptors facilitates GABA release in the hippocampus: in vitro slice and synaptosome studies. AB - Serotonin 5-hydroxytryptamine type 3 receptors (5HT3R) are Ca2+-permeant, non selective cation channels that have been localized to presynaptic terminals and demonstrated to modulate neurotransmitter release. In the present study the effect of 5-HT on GABA release in the hippocampus was characterized using both electrophysiological and biochemical techniques. 5-HT elicited a burst-like, 6- to 10-fold increase in the frequency of GABAA receptor-mediated inhibitory postsynaptic currents (IPSCs) measured with whole-cell voltage-clamp recordings of CA1 neurons in hippocampal slices. When tetrodotoxin was used to block action potential propagation, the 5-HT-induced burst of IPSCs was still observed. Stimulation of hippocampal synaptosomes with 5-HT resulted in a significant increase in the amount of [3H]GABA released by hyperosmotic saline. In both preparations, the 5-HT effect was shown to be mediated by 5HT3Rs, as it was mimicked by the selective 5HT3R agonist m-chlorophenyl biguanide and blocked by the selective 5HT3R antagonist 3-tropanylindole-3-carboxylate hydrochloride. The 5HT3R-mediated increase in GABA release was blocked by 100 microM cadmium or by omitting Ca2+ in external solutions, indicating the Ca2+-dependence of the effect. The high voltage-activated Ca2+ channel blockers omega-conotoxin GVIA and omega-conotoxin MVIIC and 10 microM cadmium had no significant effect on the 5 HT3R-mediated enhancement of GABA release, indicating that Ca2+ influx through the 5-HT3R facilitates GABA release. Taken together, these data provide direct evidence that Ca2+ entry via presynaptic 5HT3Rs facilitates the release of GABA from hippocampal interneurons. PMID- 15541892 TI - Cocaine pre-exposure produces a sensitized and context-specific c-fos mRNA response to footshock stress in the central nucleus of the AMYGDALA. AB - In recent years, there has been growing interest in the putative relationship between stress and vulnerability to relapse in former drug addicts. In animal studies aimed at exploring this relationship, it has been shown that brief exposure to intermittent footshock stress produces reliable reinstatement of drug seeking after prolonged drug-free periods. Whereas footshock reinstates drug seeking, it does not reinstate behaviors maintained by non-drug reinforcers, suggesting that prior drug experience may produce a form of sensitization within neuronal systems that mediate stress-induced reinstatement. The primary objective of the present experiments was to determine whether pre-exposure to cocaine produces a long-lasting, sensitized neuronal response to footshock stress within two brain regions known to mediate footshock-induced reinstatement; the central nucleus of the amygdala (CeA) and bed nucleus of the stria terminalis (BNST). In experiment 1, animals were injected for 7 days with cocaine (days 1 and 7 in test chambers; days 2-6 in homecages) or saline. After 21 drug-free days, they were exposed to footshock or no footshock. In experiment 2, rats were injected daily for 7 days with cocaine in one of two contexts and saline in the alternate context. After 21 drug-free days, they were given footshock either in the same context that they were given cocaine in or the alternate context. In CeA, footshock produced enhanced expression of c-fos mRNA in cocaine, but not saline, pre-exposed animals. Furthermore, this effect was gated by the environmental context in which cocaine was given; footshock only enhanced c-fos mRNA expression when it was given in a context that had previously been paired with cocaine. Although footshock induced c-fos mRNA expression in the BNST, its effects in this region were not dependent on drug history. The major findings are that a history of cocaine exposure produces sensitization to an acute stressor within CeA, and this effect is gated by environmental context. PMID- 15541893 TI - Convergence of circadian and sleep regulatory mechanisms on hypocretin-1. AB - Hypocretin is a potential regulator of sleep and wakefulness and its levels fluctuate with the day-night cycle with high levels during the animal's activity period. Whether the daily fluctuations are driven endogenously or by external light cycles is unknown. We investigated the circadian and homeostatic regulation of hypocretin in the absence of environmental light cycles. To this purpose we performed repetitive samplings of cerebrospinal fluid in rats through implanted microcannulas in the cisterna magna and determined hypocretin-1 levels by radioimmunoassay. These experiments were also performed in rats that received a lesion of the suprachiasmatic nucleus (SCN), a major pacemaker for circadian rhythms in mammals. The results showed sustained rhythmicity of hypocretin in constant dim red light in control animals. SCN-lesioned animals showed no circadian rhythms in hypocretin and mean hypocretin levels were remarkably low. The results indicate that the SCN is indispensable for rhythmicity in hypocretin and induces a daily increase in hypocretin levels during the animal's active phase. Additional sleep deprivation experiments were carried out to investigate homeostatic regulation of hypocretin. Hypocretin levels increased in response to sleep deprivation in both control and SCN-lesioned animals, demonstrating that sleep homeostatic control of hypocretin occurs independently from the SCN. Our data indicate that the circadian pacemaker of the SCN and sleep homeostatic mechanisms converge on one single sleep regulatory substance. PMID- 15541894 TI - Ultra-low dose naltrexone potentiates the anticonvulsant effect of low dose morphine on clonic seizures. AB - Significant potentiation of analgesic effects of opioids can be achieved through selective blockade of their stimulatory effects on intracellular signaling pathways by ultra-low doses of opioid receptor antagonists. However, the generality and specificity of this interaction is not well understood. The bimodal modulation of pentylenetetrazole-induced seizure threshold by opioids provide a model to assess the potential usefulness of this approach in seizure disorders and to examine the differential mechanisms involved in opioid anti- (morphine at 0.5-3 mg/kg) versus pro-convulsant (20-100 mg/kg) effects. Systemic administration of ultra-low doses of naltrexone (100 fg/kg-10 ng/kg) significantly potentiated the anticonvulsant effect of morphine at 0.5 mg/kg while higher degrees of opioid receptor antagonism blocked this effect. Moreover, inhibition of opioid-induced excitatory signaling by naltrexone (1 ng/kg) unmasked a strong anticonvulsant effect for very low doses of morphine (1 ng/kg 100 microg/kg), suggesting that a presumed inhibitory component of opioid receptor signaling can exert strong seizure-protective effects even at very low levels of opioid receptor activation. However, ultra-low dose naltrexone could not increase the maximal anticonvulsant effect of morphine (1-3 mg/kg), possibly due to a ceiling effect. The proconvulsant effects of morphine on seizure threshold were minimally altered by ultra-low doses of naltrexone while being completely blocked by a higher dose (1 mg/kg) of the antagonist. The present data suggest that ultra-low doses of opioid receptor antagonists may provide a potent strategy to modulate seizure susceptibility, especially in conjunction with very low doses of opioids. PMID- 15541895 TI - Anandamide content is increased and CB1 cannabinoid receptor blockade is protective during transient, focal cerebral ischemia. AB - The role of endocannabinoid signaling in the response of the brain to injury is tantalizing but not clear. In this study, transient middle cerebral artery occlusion (MCAo) was used to produce ischemia/reperfusion injury. Brain content of N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol were determined during MCAo. Whole brain AEA content was significantly increased after 30, 60 and 120 min MCAo compared with sham-operated brain. The increase in AEA was localized to the ischemic hemisphere after 30 min MCAo, but at 60 and 120 min, was also increased in the contralateral hemisphere. 2-Arachidonoylglycerol content was unaffected by MCAo. In a second set of studies, injury was assessed 24 h after 2 h MCAo. Rats administered a single dose (3 mg/kg) of the cannabinoid receptor type 1 (CB1) receptor antagonist SR141716 prior to MCAo exhibited a 50% reduction in infarct volume and a 40% improvement in neurological function compared with vehicle control. A second CB1 receptor antagonist, LY320135 (6 mg/kg), also significantly improved neurological function. The CB1 receptor agonist, WIN 55212 2 (0.1-1 mg/kg) did not affect either infarct volume or neurological score. PMID- 15541896 TI - Intracerebroventricular administration of anti-endothelin-1 IgG selectively upregulates endothelin-A and kappa opioid receptors. AB - Endothelin (ET) type A receptor antagonists enhance morphine-induced antinociception and restore morphine analgesia in morphine tolerant rats [Peptides 23 (2002) 1837; Peptides 24 (2003) 553]. These studies suggest that the central ET and opioid systems functionally interact. To explore this idea further, we determined the effect of i.c.v. administration of anti-ET-1 IgG (rabbit) on brain opioid receptor and ET receptor expression. Three days after implanting cannula into the lateral ventricle, male Sprague-Dawley rats were administered 10 microl (i.c.v.) of either control rabbit IgG (2.5 microg/microl) or anti-ET IgG (2.5 microg/microl) on day 1, day 3, and day 5. On day 6, animals were killed and the caudate and hippocampus collected. Anti-ET IgG had no significant effect on expression, measured by Western blots, of mu, delta or ET-B receptors, but increased kappa opioid (59%) and ET-A (33%) receptor protein expression in the caudate. [35S]-GTP-gamma-S binding assays demonstrated that anti-ET IgG decreased [D-Ala2-MePhe4, Gly-ol5]enkephalin efficacy, but not potency in the caudate. Control experiments showed that there was no detectable rabbit IgG in caudate and hippocampal samples. These results suggest that ET in the CSF negatively regulates kappa opioid and ET-A receptors in certain brain regions. These findings support the hypothesis that CSF neuropeptides have regulatory effects and further demonstrate a link between ET and the opioid receptor system. PMID- 15541897 TI - Magnetic resonance and biochemical studies during pentylenetetrazole-kindling development: the relationship between nitric oxide, neuronal nitric oxide synthase and seizures. AB - The major aim of this study was to elucidate the role of nitric oxide (NO) in the development of pentylenetetrazole (PTZ)-kindling as an animal model of primary generalized epilepsy. The daily administration of PTZ is associated with an increase in the amount of neuronal nitric oxide synthase (nNOS). NO generation was measured directly by in vivo and ex vivo electron paramagnetic resonance on rodents undergoing progressive convulsions. We found that primary generalized epilepsy is caused by NO induction during the persistent up-regulation of nNOS expression, but that NO induction is not associated with severe generalized seizures following long-term kindling phenomena after PTZ withdrawal. Morphological changes in the brain structure of rats were measured by magnetic resonance imaging during epileptic convulsions induced by repetitive administration of PTZ. Cerebellum volume for kindled rats decreased 20% but not in rats treated with the nNOS inhibitor, 3Br-7NI, suggesting that generation of NO in the cerebellum is related to decrease in cerebellum volume following PTZ kindling. PMID- 15541898 TI - T lymphocytes play a role in neuropathic pain following peripheral nerve injury in rats. AB - A catastrophic consequence of peripheral nerve injury is the development of abnormal, chronic neuropathic pain. The inflammatory response at the injury site is believed to contribute to the generation and maintenance of such persistent pain. However, the physiological significance and potential contribution of T cells to neuropathic pain remains unclear. Here we show that T cells infiltrate injured sciatic nerves following chronic constriction injury (CCI), but not uninjured nerves. Congenitally athymic nude rats, which lack mature T cells, developed a significantly reduced mechanical allodynia and thermal hyperalgesia following CCI, compared with their heterozygous littermates. To understand further the role played by different T-cell subsets, we generated polarized populations of type 1 and type 2 T cells, with different cytokine secretion profiles, from spleens of sciatic nerve-injured heterozygous rats. Passive transfer of type 1 T cells, which produce proinflammatory cytokines, into nude rats enhanced the recipients' pain hypersensitivity to a level similar to that of heterozygous donor rats. In contrast, passive transfer of polarized type 2 T cells, which produce anti-inflammatory cytokines, into heterozygous rats modestly though significantly attenuated their pain hypersensitivity. Thus, injection of type 1 and type 2 T-cell subsets produces opposing effects on neuropathic pain. These findings suggest the modulation of the T-cell immune response as a potential target for the treatment of neuropathic pain. PMID- 15541899 TI - Comparative analysis of transcriptional profiles between two apoptotic pathways of light-induced retinal degeneration. AB - Light exposure can exacerbate the condition of a variety of human retinal diseases by increasing the rate of photoreceptor cell death. How light negatively affects photoreceptor cell survival is not yet fully understood. Previous studies involving light damage models have revealed two independent apoptotic pathways: low levels of light induce retinal degeneration in the arrestin -/- mouse via constitutive activation of the phototransduction cascade, whereas strong light exposure to the retina, such as in an albino eye, elicits photoreceptor cell death via activator protein (AP-1) induction. In order to better understand the initial gene expression changes underlying light damage, dark-reared arrestin -/- and albino BALB/c mice were exposed to constant white light (2000 lux), and their retinal morphology was assessed as a function of time. The expression profiles of retinal transcripts were then compared between dark-adapted and light-exposed arrestin -/-, pigmented wild-type and BALB/c mice at a time point when morphological changes were minimal. As expected, the dark-adapted samples showed little difference in expression pattern between the three genotypes. Among the genes differentially regulated by light in BALB/c, but not arrestin -/- retinas, were c-fos and other stress-induced early response genes. In both mouse models, a marked increase in expression of the bZIP family of transcription factors was observed. Our results show a select group of unique and overlapping sets of genes induced by light in the two mouse models. These expression changes may constitute the underlying initiating events leading to the two distinct mechanisms of light damage. PMID- 15541901 TI - Gabaergic neurons with alpha2-adrenergic receptors in basal forebrain and preoptic area express c-Fos during sleep. AB - The basal forebrain (BF) contains cholinergic neurons that stimulate cortical activation during waking. In addition, both the BF and adjacent preoptic area (POA) contain neurons that promote sleep. We examined c-Fos expression in cholinergic and GABAergic neurons in the BF and POA to determine whether they are differentially active following sleep deprivation versus recovery and whether the GABAergic neurons are active during sleep. Whereas the numbers of c-Fos+ cells and proportions of c-Fos+ cells that were cholinergic were decreased, the proportions that were GABAergic were increased following sleep recovery across BF and POA nuclei. Moreover, the sleep-active GABAergic neurons were immunostained for alpha2A-adrenergic receptors. We conclude that GABAergic neurons that commonly bear alpha2-adrenergic receptors comprise sleep-active cells of the BF and POA. These GABAergic cells would be inhibited by noradrenaline (NA) released from locus coeruleus neurons during waking; they would be disinhibited through diminished NA release during drowsiness and thus become active to promote sleep by inhibiting in turn wake-promoting neurons. PMID- 15541900 TI - Pre- and post-synaptic effects of manipulating surface charge with divalent cations at the photoreceptor synapse. AB - Persistence of horizontal cell (HC) light responses in extracellular solutions containing low Ca2+ plus divalent cations to block Ca2+ currents (ICa) has been attributed to Ca2+-independent neurotransmission. Using a retinal slice preparation to record both ICa and light responses, we demonstrate that persistence of HC responses in low [Ca2+]o can instead be explained by a paradoxical increase of Ca2+ influx into photoreceptor terminals arising from surface charge-mediated shifts in ICa activation. Consistent with this explanation, application of Zn2+ or Ni2+ caused a hyperpolarizing block of HC light responses that was relieved by lowering [Ca2+]o. The same concentrations of Zn2+ and Ni2+ reduced the amplitude of ICa at the rod dark potential and this reduction was relieved by a hyperpolarizing shift in voltage dependence induced by lowering [Ca2+]o. Block of ICa by Mg2+, which has weak surface charge effects, was not relieved by low [Ca2+]o. Recovery of HC responses in low [Ca2+]o was assisted by enhancement of rod light responses. To bypass light stimulation, OFF bipolar cells were stimulated by steps to -40 mV applied to presynaptic rods during simultaneous paired recordings. Consistent with surface charge theory, the post-synaptic current was inhibited by Zn2+ and this inhibition was relieved by lowering [Ca2+]o. Nominally divalent-free media produced inversion of HC light responses even though rod light responses remained hyperpolarizing; HC response inversion can be explained by surface charge-mediated shifts in ICa. In summary, HC light responses modifications induced by low divalent cation solutions can be explained by effects on photoreceptor light responses and membrane surface charge without necessitating Ca2+-independent neurotransmission. Furthermore, these results suggest that surface charge effects accompanying physiological changing divalent cation levels in the synaptic cleft may provide a means for modulating synaptic output from photoreceptors. PMID- 15541902 TI - Behavioral, neuroendocrine and thermoregulatory actions of apelin-13. AB - As the distribution of apelinergic neurons in the brain suggests an important role of apelin-13 in the regulation of neuroendocrine processes, in the present experiments the effects of this recently identified neuropeptide on the open field activity, the hypothalamo-pituitary-adrenal (HPA) system and the body temperature were investigated. I.c.v. administration of apelin-13 (1-10 microg) to rats caused significant increases in square crossing, rearing, plasma corticosterone release and core temperature, whereas it did not influence the spontaneous motor activity during telemetric observation. To determine the mediation of the actions of apelin, a corticotropin-releasing hormone (CRH) antagonist, the nonselective dopamine antagonist haloperidol, the selective dopamine D1 receptor antagonist SCH-23390 and the nitric oxide synthase inhibitor Nomega-nitro-L-arginine-methyl ester (L-NAME) were administered to the rats. The apelin-evoked HPA activation was diminished by preadministration of the CRH antagonist, while the dopamine antagonist and L-NAME attenuated only the square crossing and rearing induced by apelin-13. To characterize the transmission of the thermoregulatory action of apelin, animals were pretreated either with L NAME, the CRH antagonist or with the cyclooxygenase inhibitor noraminophenazone. L-NAME and the CRH antagonist did not cause significant inhibition of the apelin evoked increase in core temperature, while the cyclooxygenase inhibitor, applied 30 min before peptide treatment, did not prove effective in preventing the apelin evoked thermoregulatory response, whereas when it was administered 2 h after the peptide treatment, it transiently and significantly reduced the hyperthermic response. The present data suggest that apelin-13 plays an important role in the regulation of behavioral, endocrine and homeostatic responses in the CNS, and dopamine, nitric oxide and prostaglandins seem to take part in the mediation of its effects. Since the corticosterone response could be blocked by the CRH antagonist, it is likely to be mediated through the activation of the CRH neurons. PMID- 15541903 TI - Vestibulo-reticular projections in adult lamprey: their role in locomotion. AB - This study describes the anatomical projections from vestibular secondary neurons to reticulospinal neurons in the adult lamprey and the modulation of vestibular inputs during fictive locomotion. Anatomical tracers were applied in the posterior (PRRN) and middle rhombencephalic reticular nuclei as well as to the proximal stumps of cut vestibular nerve branches to identify the neurons projecting to the reticular nuclei that were in close proximity with vestibular primary afferents. Labeled neurons were found in the intermediate (ION) and posterior (PON) octavomotor nuclei, and were more numerous on the side of the injection (around 56-87 and 101-107 for the ION and the PON, respectively). Morphologies varied but cells were mostly round or oval. Axonal projections from the PON formed a dense bundle, whereas those from the ION were less densely packed. Based on their morphology and the distribution of their projections, most vestibulo-reticular neurons were presumed to be vestibulospinal cells. Reticulospinal cells from the PRRN were recorded intracellularly in the in vitro brainstem-spinal cord preparation and large excitatory post-synaptic potentials (EPSPs) were evoked following stimulation of the ipsilateral anterior and the contralateral posterior branches of the vestibular nerves, whereas inhibitory post-synaptic potentials (IPSPs) or smaller EPSPs were elicited by stimulation of the ipsilateral posterior or of the contralateral anterior branches. During fictive locomotion, both the excitatory and the inhibitory responses displayed phasic changes in amplitude such that the amplitude of the EPSPs was minimal when the spinal cord activity switched from the ipsilateral to the contralateral side of the recorded reticulospinal cell. The IPSPs were then of maximal amplitude. We propose that this modulation could serve to reduce the influence of vestibular inputs in response to head movements during locomotion. PMID- 15541904 TI - Cell-type specific signal transduction and gene regulation via mitogen-activated protein kinase pathway in catecholaminergic neurons by restraint stress. AB - It has been demonstrated that tyrosine hydroxylase (TH) gene is easily regulated in the CNS as well as peripheral nervous systems by stressful conditions. The stimuli, such as stress or reserpine administration, significantly increased the TH gene in noradrenergic neurons in the locus ceruleus (LC), but not in dopaminergic neurons in the substantia nigra (SN). To explore the molecular mechanisms governing differential TH gene regulation in catecholaminergic cells, the present study investigated the regulation of immediate early gene (c-Fos), transcription factors (pCREB, CREB binding protein [CBP]), mitogen-activated protein (MAP) kinases (phospho-extra-cellular regulated kinase [pERK]1/2, phospho p38 MAP kinase [p-p38 MAPK], phospho-c-Jun N-terminal kinase [pJNK]) in the LC and SN in control conditions and in response to 2 h restraint stress (RS). Significant induction of c-Fos expression was observed in the LC, but not in the SN. In addition, pERK1/2 significantly increased following 2 h RS specifically in the LC, but not in the SN. No significant change was observed in p-p38 MAPK and pJNK. The expression of c-Fos and pERK1/2 preceded the upregulation of TH in the LC. Furthermore, pCREB and CBP also increased in the LC in response to 2 h RS. The induction of c-Fos prior to TH, in conjunction with the upregulation of pCREB and CBP in the LC, suggests that activator protein 1 and CRE transcription sites in the TH gene may be involved in the cell-type specific activation in the stress response, at least, by pERK1/2. PMID- 15541905 TI - Glutamate-associated plasticity in the ventral tegmental area is necessary for conditioning environmental stimuli with morphine. AB - We sought to determine if plasticity in the ventral tegmental area (VTA) of the midbrain is involved in learning to associate morphine exposure with a specific environment. For this, we tested whether activation of glutamate receptors and protein kinase A is needed for the acquisition and expression of a morphine conditioned place preference (CPP). Rats received bilateral microinjections of either the NMDA antagonist AP5 (0.48 nmol/0.3 microl), the AMPA antagonist CNQX (0.21 nmol/0.3 microl), or vehicle into the VTA prior to each of three morphine conditioning sessions. Both the AMPA and NMDA receptor antagonists blocked the development of morphine CPP when given into the VTA but not when given outside the VTA. In similar studies the protein kinase A (PKA) inhibitor, Rp-cAMPS (13 nmol/0.3 microl), blocked the acquisition of morphine CPP when given into the VTA immediately after morphine conditioning. In separate experiments, glutamate antagonists, or Rp-cAMPS, immediately prior to the preference test blocked the expression of morphine CPP when microinjected into the VTA. These data indicate that the VTA is an important site for synaptic modifications involved in the learning and memory of environmental cues predicting reward, and that glutamate input and PKA activation are crucial to this process. PMID- 15541906 TI - Investigation of intestinal absorption and disposition of green tea catechins by Caco-2 monolayer model. AB - The current study was designed to investigate the absorption mechanism and identify the possible disposition pathways of green tea catechins (GTC), including epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG) and epigallocatechin gallate (EGCG), during their absorption across small intestine by Caco-2 monolayer model. The transport of each GTC from both apical to basolateral and basolateral to apical directions was measured in the absence and the presence of MK571, an MRP inhibitor. HPLC and LC/MS were employed to identify the possible metabolites of the four GTC formed during their bidirectional transport processes. The results indicated that the four GTC showed limited transepithelial absorption with relatively small P(app) values. However, significant efflux mediated by MRP was observed during the secretion of GTC, especially the non-gallated catechins. Methylation and sulfation were the main biotransformation pathways of GTC during their secretion transport and the efflux of the related metabolites seem to be mediated by MRP. PMID- 15541907 TI - Steady-state fluorescence study on release of camptothecin from agar hydrogel. AB - The slow drug release processes from agar hydrogel were studied by steady-state fluorescence (SSF) measurement. An anticancer drug, camptothecin (CPT), was used as the release drug and the fluorescence probe for the real-time monitoring of the release processes by measuring the fluorescence intensity of the probe. The release kinetics of CPT at different temperatures was investigated. The Fickian diffusion model was adopted to fit the results and a good linear relationship could be observed between the logarithmic release fraction and the releasing time. The diffusion coefficients (D) at these conditions were thus obtained from the slopes of the fitting curves. By plotting D against temperature, the diffusion coefficient of CPT was found to obey the Arrhenius relation and the activation energy was then obtained to be 70.6 kJ mol(-1). PMID- 15541908 TI - Physicochemical properties of 2'-benzoyloxycinnamaldehyde. AB - 2'-Benzoyloxycinnamaldehyde (BCA), a derivative of 2-hydroxycinnamaldehyde demonstrated a potent antitumor effect against several human solid tumor cell lines. The physicochemical properties and degradation kinetics of BCA were investigated to support the drug-development effort. The aqueous solubility of BCA was low, and it was not considered to be hygroscopic. The degradation of BCA followed the first-order kinetics, and the pH-rate profile revealed that the degradation of BCA was governed by general acid- and specific base-catalysis as well as spontaneous hydrolysis. BCA was very unstable in basic conditions, in particular pH above 9, and found to be more stable in acidic conditions such as pH between 2 and 4. The degradation of BCA was accelerated in elevated temperature and high-ionic strength. Therefore, it was suggested that BCA should be stored in slightly acidic conditions with lowered temperature and ionic strength. PMID- 15541909 TI - Influence of selected surfactants on the tackiness of acrylic polymer films. AB - Anti-tacking agents are always necessary in polymeric film coating formulations in order to prevent substrate agglomeration. The objective of this study was to investigate the abilities of certain nonionic surfactants in a group of sorbitan ester in reducing the tackiness of the films obtained from aqueous acrylic polymer dispersions (Eudragit), compared with those of talc and glyceryl monostearate (GMS). The results from the peel tests demonstrated that GMS, Span 60 and Span 40 could significantly reduce the tackiness of both Eudragit NE 30D and Eudragit RS 30D films. The mechanisms in reducing the film tackiness were investigated by analyzing the film compositions, using attenuated total internal reflectance infrared spectroscopy (ATR-IR) and optical microscopy. The storage modulus of the films was also examined. The results indicated that GMS, Span 60, and Span 40 could reduce the film tackiness by decreasing the polymer contents at the film surfaces, resulting in a notable reduction in the contact area of the polymers between the surfaces. The use of only 5% (w/w) of either GMS, Span 60 or Span 40 in the coating formulations is enough to prevent pellet agglomeration without adverse effects on film flexibility. The pellets coated with Eudragit RS 30D/RL 30D (9:1, w/w) did not exhibit any difference in the drug release profiles when either 100% (w/w) talc or 5% (w/w) GMS was used, whereas the formulations containing Span 60 or Span 40 gave a slightly faster release rate. PMID- 15541910 TI - Study on jelly fig extract as a potential hydrophilic matrix for controlled drug delivery. AB - The principal component of aqueous extract of jelly fig (Ficus awkeotsang Makino) seeds is a pectin-type polysaccharide, gelling even at room temperature without adding any sugars, acids or ions. The objective of this study was to evaluate jelly fig extract (JF) as a matrix base for sustained release tablets. Drug release profile from JF tablet was examined using theophylline as a model drug, compared with those from USP graded pectin (USP-P). Release profile from JF tablet was a sustained release pattern and not affected by pH of medium. USP-P tablet showed a similar release profile of JF tablet, however, the release mechanisms differed. Matrix erosion studies revealed that the percentage of drug released from USP-P tablet was proportional to that of matrix eroded. On the other hand, JF tablet was eroded up to 50% of matrix for 4h and showed a constant value thereafter. According to water uptake studies, JF tablet showed an initial burst swelling followed by slow water uptake, suggesting diffusion-controlled kinetics in later phase. Moreover, theophylline release rate from JF tablet was modified by drug content in the tablet, increasing with decrease in drug amount. These findings indicated JF was a potential hydrophilic matrix for controlled drug delivery. PMID- 15541911 TI - Clinical pharmacokinetic and pharmacodynamic evaluation of transdermal drug delivery systems of salbutamol sulfate. AB - Transdermal drug delivery formulation containing 5 mg/patch of salbutamol sulfate (SS), providing an input rate of 100 microg/h of SS was formulated and subjected for pharmacokinetic and pharmacodynamic evaluation in moderately asthmatic patients (n=6). A linear correlation was observed between cumulative amount of drug diffused in vitro and cumulative AUC0-t of serum concentration-time curve (R2=0.99). A steady-state serum concentration of 2.87+/-0.1 ng/ml (per milligram dose) was attained after an initial lag period of 4.67+/-1.03 h. The elimination half-life, clearance rate and elimination rate constant was 3.35+/-1.07 h, 256.12+/-3.55 ml/min and 0.24+/-0.09 h(-1), respectively. The mean forced expiratory volume in one minute (FEV1) of the patients was 2.2+/-0.14l during steady state. The pharmacokinetic results correlated well with the FEV1 response of patients. PMID- 15541912 TI - Process characteristics and compaction of spray-dried emulsions containing a drug dissolved in lipid. AB - The objective of the present study is to prepare directly compressible powders, containing a poorly water-soluble drug dissolved in medium-chain triglycerides (MCT), by spray drying o/w-emulsions in a pilot plant spray dryer. In addition to the lipid phase, the emulsions contained a water-soluble carrier (a sugar), a water-insoluble carrier (magnesium alumino metasilicate) and a combined emulsifier and film-forming agent (gelatine). A factorial design was used to investigate the effect of formulation variables on the spray drying process and powder properties. The factors varied were soluble carrier type (trehalose or mannitol), insoluble carrier particle size distribution (granular or fine powder) and amount of lipid phase in the emulsion (low or high). Compressibility and compactibility of the spray-dried emulsions were mainly affected by the content of lipid in the powders and decreased on increasing the amount of lipid. Increasing the particle size of the insoluble carrier decreased spray drying process yield and lipid encapsulation efficiency whereas compactibility and handling properties were improved. Incorporation of a soluble carrier becoming amorphous on spray drying resulted in tablets with an increased mechanical strength compared to powders containing a crystalline soluble carrier. PMID- 15541913 TI - Enhanced transdermal delivery of atenolol from the ethylene-vinyl acetate matrix. AB - To enhance transdermal delivery of atenolol, ethylene-vinyl acetate (EVA) matrix of drug containing penetration enhancer was fabricated. Effect of penetration enhancer on the permeation of atenolol through the excised rat skin was studied. Penetrating enhancers showed the increased flux probably due to the enhancing effect on the skin barrier, the stratum corneum. Among enhancers used such as glycols, fatty acids and non-ionic surfactants, polyoxyethylene 2-oleyl ether showed the best enhancement. For the controlling transdermal delivery of atenolol, the application of EVA matrix containing permeation enhancer could be useful in the development of transdermal drug delivery system. PMID- 15541914 TI - Development of lidocaine gels for enhanced local anesthetic action. AB - In relieving local pains, lidocaine, one of ester type local anesthetics, has been used. To develop the lidocaine gels of enhanced local anesthetic effects, hydroxypropyl methylcellulose (HPMC) based bioadhesive polymer gel containing an enhancer was formulated. As the drug concentration in the gels increased up to 3%, the permeation rate of drug linearly increased, thereafter reaching a plateau. As the temperature of surrounding solutions increased, the permeation of drug increased. The activation energy of drug permeation was 3.29 kcal/mol for lidocaine. The permeation rate of drug through skin was studied using various enhancers, such as glycols, non-ionic surfactants, and bile salts. Among the enhancers studied, diethylene glycol showed the greatest enhancing effects on drug permeation through skin. The analgesic activity was examined using a tail flick analgesimeter. In the area under the efficacy curve (AUEC) of the rat-tail flick tests, lidocaine gel containing diethylene glycol showed about 3.89-fold increase in analgesic activity compared with the control. The addition of vasoconstrictor in the gels prolonged the analgesic effects. The result of this study supports that the bioadhesive gel with efficient anesthetic effect could be developed using HPMC with combination of enhancer and vasoconstrictor. PMID- 15541915 TI - An investigation into the predictive value of cascade impactor results for side effects of inhaled salbutamol. AB - The aim of this study was to compare the Multistage Liquid Impinger (MSLI) and the Andersen Cascade Impactor (ACI) with respect to their power to predict differences in side effects of salbutamol delivered by a dry powder inhaler. Three preparations with the same nominal dose and the same inhaler device but generating aerosols with different aerodynamic particle size distributions were administered to six healthy volunteers in a randomized, placebo-controlled, four way crossover study. Cumulative doses from 400 up to 1600 microg were given. The serum potassium level (K+-serum) and the heart rate (HR) were measured at baseline and 15 min after each dose. Both the MSLI and ACI showed large differences between the aerodynamic particle size distributions of the three preparations. The decrease in K+-serum revealed significant differences between the three active preparations and was significant for doses of 800 microg and higher. The HR results showed differences between the active preparations only at a nominal dose of 1600 microg and only for the preparation with the highest fine particle dose (FPD) compared to the other two preparations. The K+-serum appears to be a more sensitive measure for side effects than the HR. In vivo-in vitro correlations (IVIVCs) were established between the amounts of salbutamol deposited on the various cumulative impactor stages and the K+-serum. The best IVIVCs were obtained in the FPD range, resulting in correlation coefficients of at least 0.78. It is concluded that cascade impactor results in the FPD range of the MSLI as well as the ACI correlate well with the K+-serum. Cascade impactor analysis thus provides a clinically meaningful tool in the development and the quality control of salbutamol inhalation powders. PMID- 15541916 TI - Enteric-coated layered double hydroxides as a controlled release drug delivery system. AB - Layered double hydroxides (LDHs) or so-called anionic clays consist of cationic brucite-like layers and exchangeable interlayer anions. Because of their biocompatibility, these layered inorganic solids can be used as host materials to create drug-LDH host-guest supramolecular structures. Because of the basicity of LDHs however, LDHs as drug delivery system will be limited for use in the stomach where pH is 1.2. A core-shell material has been prepared therefore in this work. A non-steroidal anti-inflammatory drug, Fenbufen-intercalated LDHs as the core was coated with enteric polymers, Eudragit S 100 or Eudragit L 100 as a shell, giving a composite material which shows controlled release of the drug under in vitro conditions which model the passage of a material through the gastrointestinal tract. PMID- 15541917 TI - Dissolution test as a surrogate for quality evaluation of rifampicin containing fixed dose combination formulations. AB - The present investigation was aimed at developing a dissolution methodology to predict in vivo performance of rifampicin containing fixed dose combination (FDC) products. Six FDC formulations were used in this study, of which four had passed bioequivalence while two failed. Dissolution studies were conducted at agitation intensity of 30-100 rpm as a measure of hydrodynamic stress and at pH media corresponding to gastric and intestinal conditions. Formulations showed variable dissolution at different conditions and dissolution at 50 rpm was most sensitive and differentiated the release profiles of rifampicin under various pH conditions. It was possible to predict in vivo performance of rifampicin from FDCs when in vitro rate and extent of release at various pH was correlated with site, pH and concentration dependent absorption of rifampicin along with gastric emptying time. It was also seen that dissolution conditions recommended in USP for different types of FDCs were insensitive for the formulation changes. Based on this comprehensive evaluation, a decision tree is proposed which will act as a guideline for quality evaluation of FDC products and also will provide a fundamental knowledge for optimization of formulations failing in dissolution studies. PMID- 15541918 TI - A study of liposome formation using a solution (isoperibol) calorimeter. AB - A solution (isoperibol) calorimeter has been employed to study the process of formation of phospholipid vesicles from natural and synthetic phospholipid films. Phospholipid films were hydrated in the solution calorimeter at temperatures exceeding the main phospholipid phase transition temperature, with continuous agitation to ensure conversion of the hydrating bilayers into multilamellar liposomes. It was seen that retention of chloroform in phospholipid films altered the apparent enthalpy change of vesicle formation to a far greater extent than would be expected from the contribution of the enthalpy of solution of chloroform; this indicates that chloroform alters the hydration process of the lipid. The overall measured enthalpy change for the formation of egg phosphatidylcholine vesicles was exothermic, whilst that for dimyristoylphosphatidylcholine was endothermic. This difference, it is suggested, results from the influence of the hydrocarbon chains mostly on the hydration process and also on the process of vesicle formation. PMID- 15541919 TI - Determining the critical relative humidity for moisture-induced phase transitions. AB - A new method to determine the onset relative humidity for a glass transition and crystallization processes in amorphous or partially amorphous materials was developed using dynamic gravimetric vapor sorption (DVS). Water vapor can act as a plasticizing agent in amorphous materials, thus lowering the glass transition temperature below room temperatures. Additional water sorption can lead to a crystallization event below the glass transition temperature. On spray-dried lactose the glass transition RH and crystallization RH values were 30 and 58% at 25 degrees C, respectively. Glass transition and crystallization RH values were also measured at 5, 15, 25, 35, and 45 degrees C on a spray-dried salbutamol sulfate sample. The glass transition RH values for the salbutamol sulfate sample ranged from 64.5% RH (5 degrees C) to 32.8% RH (45 degrees C) while the crystallization RH values ranged from 81.0% RH (5 degrees C) to 50.4% RH (45 degrees C). The results clearly show that the glass transition and crystallization humidity values decrease as the sample temperature increases. PMID- 15541920 TI - Search for technological reasons to develop a capsule or a tablet formulation with respect to wettability and dissolution. AB - Proquazone, a poorly wettable compound, was used as a model drug in the search for reasons to develop a capsule or tablet formulation. The capsules were filled with proquazone as active ingredient, with lactose monohydrate (200 mesh) as filler and with magnesium stearate as lubricant. The tablet was made out of a granulate as internal phase which consisted of proquazone as active ingredient, lactose as filler, corn starch as disintegrant and PVP as a binding agent. The external phase consisted of magnesium stearate and corn starch. The concentration of proquazone in the capsule and in the tablet formulation was varied. The capsule formulations showed a significantly slower dissolution of the drug substance than the tablet formulations especially for a high-drug load. Independently of the drug load, only the tablet formulation showed a high dissolution rate. Thus, concerning drug load, only the tablet formulations showed to be robust. It became clear that proquazone needs to be formulated as a granulate or a tablet to achieve a fast dissolution rate. Thus, a poorly wettable drug, especially when it is found in high concentrations, can have direct impact on the decision to develop a tablet or a capsule formulation. PMID- 15541921 TI - Uptake characteristics of mannosylated and fucosylated bovine serum albumin in primary cultured rat sinusoidal endothelial cells and Kupffer cells. AB - The purpose of this study is to delineate uptake characteristics of mannosylated and fucosylated proteins in primary cultured sinusoidal endothelial cells and Kupffer cells. In cultured sinusoidal endothelial cells, uptake of mannosylated and fucosylated bovine serum albumin (BSA) was significantly inhibited by excess mannosylated and fucosylated BSAs but not by galactosylated BSA, suggesting that both glycosylated proteins might be primarily taken up via mannose receptors. In cultured Kupffer cells, uptake of fucosylated BSA was significantly inhibited by excess galactosylated BSA as well as mannosylated and fucosylated BSAs, although that of mannosylated BSA was inhibited only by mannosylated and fucosylated BSAs. This suggests that uptake of fucosylated BSA by Kupffer cells might be mediated by both Kupffer cell lectin (fucose receptor) and mannose receptor. On the other hand, in vivo hepatic uptake of fucosylated BSA was inhibited to a greater extent by GdCl3 pretreatment than that of mannosylated BSA. Based on in vitro and in vivo experiments, it was concluded that fucosylated BSA is more Kupffer cell selective because it exhibited a lower sinusoidal endothelial cell uptake than mannosylated BSA. PMID- 15541922 TI - Uptake of folate-conjugated albumin nanoparticles to the SKOV3 cells. AB - The bovine serum albumin nanoparticles (BSANPs) were prepared by a coacervation method and chemical cross-linking with glutaraldehyde. Furthermore, the BSANPs were reacted with the activated folic acid to conjugate folate via amino groups of the BSANPs, to improve their intracellular uptake to target cells. The nanoparticles were apparently spherical with diameters less than 150 nm and their average diameter was 70 nm by a transmission electron microscope (TEM) and a laser light scattering particle analyzer, respectively. An extent of folate conjugation with the BSANPs was 169 micromol/g BSA by spectrophotometric analysis. Cell uptake studies were carried out in SKOV3 cells (human ovarian cancer cell line) using fluorescein isothiocyanate labeled nanoparticles. The extent of BSANPs taken up by the cells was measured with a fluorescence spectrophotometer. The nanoparticles were taken up to the cells and levels of binding and uptake were increased with the time of incubation until 4 h. The levels of folate-conjugated BSANPs were higher than those of BSANPs and saturable. The association of folate-conjugated BSANPs to SKOV3 cells was inhibited by an excess amount of folic acid, suggesting that the binding and/or uptake were mediated by the folate receptor. These results implied that the folate-conjugated BSANPs might be useful as a drug carrier system to deliver drugs into the cells expressing folate receptor. PMID- 15541923 TI - Nisin-loaded poly-L-lactide nano-particles produced by CO2 anti-solvent precipitation for sustained antimicrobial activity. AB - Nisin-loaded poly-L-lactide (PLA) nano-particles were fabricated by processing protein/polymer organic solutions by semi-continuous compressed CO2 anti-solvent precipitation. Preliminary solubility studies were carried out for an optimised selection of organic solvent mixtures leading to preparation of protein/polymer solutions. The particles were prepared by processing 50:50 dimethylsulfoxide/dichloromethane mixtures containing 1% polymer (w/v) and 5 or 20% nisin (nisin/polymer, w/w). Proper operative conditions (organic solution injection rate, precipitation temperature and gas pressure, CO2 flow rate, washing time, etc.) were set up to yield production reproducibility, high product recovery (over 70%) and high drug loading (over 95% of the recovered protein). Scanning electron microscopy demonstrated that spherical, smooth surfaced particles were produced. Light scattering showed that the particle size was in the range of 200-400 nm and the products were characterised by narrow polydispersity. In vitro release studies showed that the protein is slowly released throughout 1000 h. However, the release was slower as the salt concentration and the pH of the release buffer increased. Solubility investigations suggested that the observed differences in protein release rate out of nano-particles was attributable to the protein interaction with the polymer which was found to increase as the pH or the salt concentration increased in the release buffer. In vitro studies carried out by nano-particle incubation in medium containing Lactobacillus delbrueckeii showed that nisin was released in the active form and the antibacterial activity was maintained up to 45 days incubation. PMID- 15541926 TI - Cadaveric donors. AB - The diagnosis of brain death describes the final catastrophic state of a person for whom, except for ventilators and pharmacology, death would surely have already intervened. Although we think of death as an ending, if the patient becomes an organ donor it is also a beginning. For, at the moment we lose one patient, we potentially gain others in the fortunate recipients of viable grafts. PMID- 15541927 TI - Living donor surgery: overview of surgical and anesthesia issues. AB - Organ transplantation occupies the center stage in the treatment of many forms of end-stage organ disease. When the limits of conventional medical care are exhausted, bridging therapies, cadaveric transplantation, and posttransplant medical care come to the fore. Living donor transplantation has grown out of the numerical and immunosuppression limitations of this process. Living donor transplantation medicine and surgery encompass two of the most fascinating and compelling social and ethical dilemmas of modern health care. This article provides an overview of medical and ethical concerns for those who decide to become living donors and those who care for them in the perioperative period. PMID- 15541928 TI - Kidney transplantation: recent developments and recommendations for anesthetic management. AB - Kidney transplantation is the treatment of choice for patients with end-stage renal disease. After receiving a transplant, survival rates are higher and comorbidities may resolve. As a consequence, more patients with significant comorbidities such as advanced cardiovascular disease will present for transplantation. This review highlights commonly encountered issues in patients undergoing kidney transplantation and recommendations are made for their anesthetic management. PMID- 15541929 TI - Pancreas and islet cell transplantation. AB - Transplantation of the pancreas or islet cells constitutes surgical treatment for patients with type 1 diabetes mellitus. Pancreas transplantation is now an established procedure for the surgical treatment of diabetes mellitus. Islet cell transplantation has the potential to be the procedure of choice once it becomes more routine because of the minimal surgery involved. Included in this chapter are the pathophysiology of diabetes, rationale for transplantation, and the surgical procedure itself. PMID- 15541930 TI - Anesthetic management for small bowel transplantation. AB - Small bowel transplantation is quickly becoming the therapy of choice for short gut syndrome. Improvements in surgical techniques, immunosuppressants, and anesthetic management of patients have allowed this procedure to become the standard of care for patients with irreversible intestinal failure who are unable to continue total parenteral nutrition therapy. This article discusses the evaluation process for candidate selection, including work-up for vascular access and hypercoagulability, intraoperative management of patients (including appropriate monitoring, hemodynamic changes, fluid management, surgical techniques, and effects of immunosuppressants), and patient survival and outcome. PMID- 15541931 TI - Anesthesia for liver transplant surgery. AB - Liver transplantation offers patients with liver disease an optimal chance for long-term survival. Current indications, preoperative assessment, patient selection, intraoperative anesthetic management and outcomes are described. The management of special situations, including retransplantation, pediatric transplantation, and fulminant hepatic failure are also reviewed. The success of liver transplantation has led to increased demand. This demand, coupled with a nonexpanding supply of deceased donor organs, has resulted in a shortage of grafts and prolonged waiting times. Novel solutions using segmental liver grafts from living donors, and the challenges associated with this approach, are discussed. PMID- 15541932 TI - Surgical advances in liver and bowel transplantation. AB - Liver and intestinal transplantation are currently the treatments of choice for life-threatening hepatic and gastrointestinal failure. These technologies have evolved through contributions from the fields of immunology, anatomy, physiology, surgery, anesthesiology, critical care, ethics, epidemiology, and public health. Transplantation now accounts for the treatment of over 5,000 recipients per year who are in a state of organ failure. The available donor population, however, is not increasing to meet the demands of the faster growing recipient population. This discrepancy has led to the rapid development of novel strategies that require critical evaluation to build on the success rates in recent years. This article presents the most salient advances in liver and intestinal transplantation in the last 15 years. PMID- 15541933 TI - Anesthetic considerations for multivisceral transplantation. AB - Multivisceral transplantation is defined as the en bloc transplantation of three or more abdominal organs. Although multivisceral transplant recipients do not yet enjoy the same survival rates as renal and liver transplant recipients, this procedure can be life saving and has shown improvements in survival rates over time. Advances in immunosuppression hold promise for the future of multivisceral transplantation. PMID- 15541934 TI - Cardiac transplantation. AB - Cardiac transplantation is a proven, accepted mode of therapy for selected patients with end-stage heart failure, but the inadequate number of suitable donor hearts available ultimately limits its application. This chapter reviews adult cardiac transplantation, with an emphasis on the anesthetic considerations of the heart transplant operation itself. PMID- 15541935 TI - Anesthetic implications for lung transplantation. AB - Anesthetic challenges regarding lung transplantation are related to the expanded spectrum of diseases for which lung transplantation is offered and to the interval changes in health status likely to occur as patients wait longer for an organ to become available. Particular attention to avoiding or reducing the impact of increases in pulmonary vascular resistance and right heart failure are important and may necessitate cardiopulmonary bypass. Intraoperative and postoperative ventilator management should account for differences in pulmonary compliance after the new lung is implanted. Minimizing intravenous fluids without compromising end organ perfusion may avoid or reduce postoperative respiratory insufficiency. PMID- 15541936 TI - Surgical advances in heart and lung transplantation. AB - The successes of thoracic transplantation have led to the expansion of indications and a subsequent growth in demand for a short supply of organs. In response to this disparity, the criteria for organ donation have been liberalized. Despite these difficult challenges, with advances in surgical techniques and perioperative care of both the donor and recipient, outcomes have continued to improve over time. This article focuses on the more recent surgical advances in donor selection and management, procurement and implantation, and the impact of these advances on patient outcome. PMID- 15541937 TI - Anesthetic management of the pediatric patient undergoing solid organ transplantation. AB - Solid organ transplantation is now routinely performed at many institutions. Pediatric organ recipients present difficult challenges to pediatric anesthesiologists. Physiologic, anatomic, and pharmacologic derangements in this population may make both the surgical procedure and the anesthetic management complicated. This article presents an overview of the unique problems and the strategies to solve them in this population. PMID- 15541938 TI - The transplant recipient for nontransplant surgery. AB - The transplant recipient has traded a life-threatening illness for a chronically immunosuppressed state. Subsequent anesthetic management for non-transplant surgical procedures may be challenging. The anesthesia provider must be aware of the degree of post-transplant organ dysfunction and alter anesthesia techniques accordingly. This article reviews the anesthetic concerns for patients who have undergone a variety of organ transplants. PMID- 15541939 TI - Bridges to transplantation. AB - The definitive treatment of patients with acute liver failure is liver transplantation. Organ Procurement and Transplantation Network status 1 patients receive priority, but the median waiting time in 2002 was 11 days. Patients who are small or have an unusual blood type are expected to wait even longer. Because cerebral edema and death may occur before a liver is available, numerous methods of bridging patients to transplantation by artificial means have been proposed. To date, no system of hepatic support has been proven effective at delaying the onset of cerebral edema in controlled trials. PMID- 15541940 TI - Xenograft transplantation. AB - Interest in xenotransplantation has increased because conventional organ transplantation has been limited by a shortage of human organs. Although xenotransplantation could alleviate the existing and anticipated need for tissues and organs, the application is hindered by various biologic obstacles. This article reviews the basis for the demand for xenotransplantation, the obstacles to clinical application, and potential approaches to overcoming those obstacles. PMID- 15541941 TI - Cellular transplantation. AB - This article reviews the scant literature that exists describing the interface between anesthesiologists and marrow donors and islet recipients, introduces the issues surrounding future stem cell transplantation technologies, and describes pretranslational cell transplant applications that are closest to clinical trials. PMID- 15541942 TI - Bayesian representation learning in the cortex regulated by acetylcholine. AB - A brain needs to detect an environmental change and to quickly learn internal representations necessary in a new environment. This paper presents a theoretical model of cortical representation learning that can adapt to dynamic environments, incorporating the results by previous studies on the functional role of acetylcholine (ACh). We adopt the probabilistic principal component analysis (PPCA) as a functional model of cortical representation learning, and present an on-line learning method for PPCA according to Bayesian inference, including a heuristic criterion for model selection. Our approach is examined in two types of simulations with synthesized and realistic data sets, in which our model is able to re-learn new representation bases after the environment changes. Our model implies the possibility that a higher-level recognition regulates the cortical ACh release in the lower-level, and that the ACh level alters the learning dynamics of a local circuit in order to continuously acquire appropriate representations in a dynamic environment. PMID- 15541943 TI - Criteria for exponential [corrected] stability of Cohen-Grossberg neural networks. AB - In this paper, the Cohen-Grossberg neural network models without and with time delays are considered. By constructing several novel Lyapunov functionals, some sufficient criteria for the existence of a unique equilibrium and global exponential stability of the network are derived. These results are fairly general and can be easily verified. Besides, the approach of the analysis allows one to consider different types of activation functions, including piecewise linear, sigmoids with bounded activations as well as C1-smooth sigmoids. In the meantime, our approach does not require any symmetric assumption of the connection matrix. It is believed that these results are significant and useful for the design and applications of the Cohen-Grossberg model. PMID- 15541944 TI - Global exponential stability of cellular neural networks with time-varying coefficients and delays. AB - In this paper, a class of cellular neural networks with time-varying coefficients and delays is considered. By constructing a suitable Liapunov functional and utilizing the technique of matrix analysis, some new sufficient conditions on the global exponential stability of solutions are obtained. The results obtained in this paper improve and extend some of the previous results. PMID- 15541945 TI - A theory of epineuronal memory. AB - How can a brain maintain stable memories and behaviors when its underlying electrical and chemical structures are constantly changing? We investigate this stability problem thinking that the state variables (e.g. voltages, ionic currents, etc.) are governed by a complex system that itself is closely regulated. Regulation of the network is through its operating environment, which is described by parameters. We study a standard neural network model, but one whose parameters are governed by a mnemonic landscape function. Parameter configurations are attracted to local maxima of this landscape, which represent memorized parameter configurations. The operating environment changes slowly (relative to the state variables) guided by the mnemonic landscape function, and this provides a quasi-static operating environment for the network. We describe shaping the mnemonic landscape, how it acts as a probability density function to guide slow parameter dynamics, and how the parameters shape the network output. Simulations of memory registration, activity patterns, and sequential recall are described. PMID- 15541946 TI - Sparse distributed memory using N-of-M codes. AB - An analysis is presented of a sparse distributed memory (SDM) inspired by that described by Kanerva [Kanerva, P. (1988). Sparse distributed memory. Cambridge, MA: MIT Press] but modified to facilitate an implementation based on spiking neurons. The memory presented here employs sparse binary N-of-M codes, unipolar binary synaptic weights and a simple Hebbian learning rule. It is a two-layer network, the first (fixed) layer being similar to the 'address decoder' in Jaeckel's [Jaeckel, L.A. (1989). A class of designs for a sparse distributed memory. RIACS Technical Report 89.30, NASA Ames Research Centre] 'hyperplane' variant of Kanerva's SDM and the second (writeable) 'data store' layer being a correlation matrix memory as first proposed by Willshaw et al. [Willshaw, D. J., Buneman, O.P., & Longuet-Higgins, H.C. (1969). Non-holographic associative memory. Nature, 222, 960-962]. The resulting network is shown to have good storage efficiency and is scalable. The analysis is supported by numerical simulations and gives results that enable the configuration of the memory to be optimised for a range of noiseless and noisy environments. PMID- 15541947 TI - Feedback error learning and nonlinear adaptive control. AB - In this paper, we present our theoretical investigations of the technique of feedback error learning (FEL) from the viewpoint of adaptive control. We first discuss the relationship between FEL and nonlinear adaptive control with adaptive feedback linearization, and show that FEL can be interpreted as a form of nonlinear adaptive control. Second, we present a Lyapunov analysis suggesting that the condition of strictly positive realness (SPR) associated with the tracking error dynamics is a sufficient condition for asymptotic stability of the closed-loop dynamics. Specifically, for a class of second order SISO systems, we show that this condition reduces to K2D > K(P) where K(P) and K(D) are positive position and velocity feedback gains, respectively. Moreover, we provide a 'passivity'-based stability analysis which suggests that SPR of the tracking error dynamics is a necessary and sufficient condition for asymptotic hyperstability. Thus, the condition K2D > K(P) mentioned above is not only a sufficient but also necessary condition to guarantee asymptotic hyperstability of FEL, i.e. the tracking error is bounded and asymptotically converges to zero. As a further point, we explore the adaptive control and FEL framework for feedforward control formulations, and derive an additional sufficient condition for asymptotic stability in the sense of Lyapunov. Finally, we present numerical simulations to illustrate the stability properties of FEL obtained from our mathematical analysis. PMID- 15541948 TI - Fast exact leave-one-out cross-validation of sparse least-squares support vector machines. AB - Leave-one-out cross-validation has been shown to give an almost unbiased estimator of the generalisation properties of statistical models, and therefore provides a sensible criterion for model selection and comparison. In this paper we show that exact leave-one-out cross-validation of sparse Least-Squares Support Vector Machines (LS-SVMs) can be implemented with a computational complexity of only O(ln2) floating point operations, rather than the O(l2n2) operations of a naive implementation, where l is the number of training patterns and n is the number of basis vectors. As a result, leave-one-out cross-validation becomes a practical proposition for model selection in large scale applications. For clarity the exposition concentrates on sparse least-squares support vector machines in the context of non-linear regression, but is equally applicable in a pattern recognition setting. PMID- 15541949 TI - An on-line algorithm for creating self-organizing fuzzy neural networks. AB - This paper presents a new on-line algorithm for creating a self-organizing fuzzy neural network (SOFNN) from sample patterns to implement a singleton or Takagi Sugeno (TS) type fuzzy model. The SOFNN is based on ellipsoidal basis function (EBF) neurons consisting of a center vector and a width vector. New methods of the structure learning and the parameter learning, based on new adding and pruning techniques and a recursive on-line learning algorithm, are proposed and developed. A proof of the convergence of both the estimation error and the linear network parameters is also given in the paper. The proposed methods are very simple and effective and generate a fuzzy neural model with a high accuracy and compact structure. Simulation work shows that the SOFNN has the capability of self-organization to determine the structure and parameters of the network automatically. PMID- 15541950 TI - Interactive detection and visualization of breast lesions from dynamic contrast enhanced MRI volumes. AB - Mammography is currently regarded as the most effective and widely used method for early detection of breast cancer, but recently its sensitivity in certain high risk cases has been less than desired. The use of Dynamic Contrast Enhanced Magnetic Resonance Imaging (DCE-MRI) has gained considerable attention in the past 10 years, especially for high risk cases, for smaller multi-focal lesions, or very sparsely distributed lesions. In this work, we present an interactive visualization system to identify, process, visualize and quantify lesions from DCE-MRI volumes. Our approach has the following key features: (1) we determine a confidence measure for each voxel, representing the probability that the voxel is part of the tumor, using a rough goodness-of-fit for the shape of the intensity time curves, (2) our system takes advantage of low-cost, readily available 3D texture mapping hardware to produce both 2D and 3D visualizations of the segmented MRI volume in near real-time, enabling improved spatial perception of the tumor location, shape, size, distribution, and other characteristics useful in staging and treatment courses, and (3) our system permits interactive manipulation of the signal-time curves, adapts to different tumor types and morphology, thus making it a powerful tool for radiologists/physicians to rapidly assess probable malignant volumes. We illustrate the application of our system with four case studies: invasive ductal cancer, benign fibroadenoma, ductal carcinoma in situ and lobular carcinoma. PMID- 15541951 TI - Integration of color and boundary information for improved region of interest identification in electron magnetic resonance images. AB - A Windows-based, object-oriented application system for segmentation and analysis of electron magnetic resonance (EMR) images is described. The integrated system is developed for better recognition of regions of interest (ROI) in murine EMR images. The system combines the clustering method of color segmentation with boundary detection, for efficient segmentation of regions of interest in EMR images. Initially, the red/green/blue (RGB) color space is converted into spherical coordinates transform (SCT) space. Color quantization is then achieved by center split algorithm applied on the color dimensions of the SCT space. Subsequently, Laplacian boundary detection operator is used to extract the contours of the ROI from the variegated coloring information. The system is implemented in Visual C++ and tested on temporal EMR color images of mouse. The system performs well giving perceptually reasonable segmentation of tumor, kidney and bladder of the mouse image. Experimental results with extensive set of EMR color images demonstrate the efficacy of the system developed. PMID- 15541952 TI - Noise reduction in spine videofluoroscopic images using the undecimated wavelet transform. AB - Videofluoroscopy permits using sequences of low quality images to study the spine movement. In this work the problem of enhancing the quality of these images is considered in order to facilitate the extraction of kinematic parameters. The method is based on the undecimated wavelet transform and on a preliminary training of a sub-set of images. The anatomical features are preserved using a mask. Key element of the method is its fast and automated implementation. The concept of improving the extraction of kinematic parameters by improving the image representation instead of the technique to extract these is also innovative. The technique has been tested on two sequences of images and the results demonstrates that the method enhances images not related with training sub-set. PMID- 15541953 TI - Speckle reducing anisotropic diffusion for 3D ultrasound images. AB - This paper presents an approach for reducing speckle in three dimensional (3D) ultrasound images. A 2D speckle reduction technique, speckle reducing anisotropic diffusion (SRAD), is explored and extended to 3D. 3D SRAD is advantageous in that, like 2D SRAD, it keeps the advantages of the conventional anisotropic diffusion and the traditional speckle reducing filter, the Lee filter, by exploiting the instantaneous coefficient of variation (ICOV). Besides, 3D SRAD uses 3D information; thus it overcomes the shortcoming of the 2D technique that only uses 2D information. The algorithm of 3D SRAD is presented in the continuous domain as well as in the discrete domain. Experiments have been performed on both synthetic and real 3D ultrasound images and the experimental results were compared with those obtained by 3D anisotropic diffusion and the 3D Lee filter. The experimental results show that the quality of the 3D SRAD for speckle reduction in 3D ultrasound images improves upon that of 3D anisotropic diffusion and 3D Lee filter in terms of edge preservation and the smoothness of homogenous regions. PMID- 15541954 TI - Acceleration of 3D, nonlinear warping using standard video graphics hardware: implementation and initial validation. AB - Thin Plate Spline (TPS) transformations are used in many medical imaging algorithms, but are time-prohibitive for iterative or interactive use. We have utilized current generation consumer 3D graphics cards to accelerate the application of the TPS nonlinear transformation by combining hardware accelerated 3D textures, vertex shaders and trilinear interpolation. Our hardware accelerated algorithm warped a 512x512x173 Computed Tomography (CT) dataset in 2.3 s, and the same study, scaled to 256x256x173, in 0.5 s, using a set of 92 landmarks. An accelerated software implementation of the TPS transformation warped the 512x512x173 study in 32.9 s and the 256x256x173 study in 9.5 s. Subtracted images were used for qualitative analysis of the warp and the Mutual Information (MI) and Sum of Absolute Difference (SAD) similarity metrics were used to quantitatively compare the output of the hardware accelerated algorithm to that of the software algorithm. The hardware accelerated algorithm provided a 7-65 times performance increase when compared to the software algorithm, and produced output of comparable quality (MI > 300 and SAD < 1.75%). PMID- 15541956 TI - The human genome project and the discovery of genetic determinants of cancer susceptibility. AB - The Human Genome Project has recently provided a great deal of information on the sequence that comprises the human genome. We are now in the process of structuring and deciphering the 3 x 10(9) base sequence in order to gain insights into its functional role. Several efforts are focusing on the search for DNA sequence variations underlying common/complex diseases that constitute a real burden in terms of public health measures. As expected, the genetic architecture of these complex traits, shows tremendous complexity, and the discovery and characterisation of susceptibility alleles constitute a real challenge for the geneticist. Conceptual and experimental genetic approaches aimed at dissecting the molecular features of susceptibility genes, in particular those predisposing to cancer, are outlined and discussed in this review. PMID- 15541957 TI - Transcription profiling of gene expression in drug discovery and development: the NCI experience. AB - Transcript profiling, using microarray or other analogous technologies, to query on a large-scale the expression of genes in tumours or their derivative cell lines has numerous potential uses in oncology drug discovery and development. Characterisation of genes expressed in tumours may allow tumours to be separated into subsets defining subtypes that have a distinctive pathway utilisation. The molecular entities comprising the pathways which distinguish one disease subset from another then become potential candidate drug targets. Alternatively, gene expression patterns may be correlated with the degree of antiproliferative effect of candidate drug leads. This can reveal aspects of the drug's action that could serve to provide a further basis for benchmarking the generation of analogues or provide important information about pathways potentially modulated by the drug in achieving cytotoxicity. New information is emerging that the expression of drug transport-related molecules is a major variable that can be usefully explored using gene expression data, and the features promoting successful drug handling by the tumour cell may be an additional variable which can be illuminated by gene expression studies. PMID- 15541958 TI - Improving the selectivity of cancer treatments by interfering with cell response pathways. AB - The cellular response to the stress induced by treatment with anticancer agents is a key determinant of drug activity. A pivotal role in this response is played by checkpoint proteins that control the normal passage of cells through the cell cycle. There is evidence that cancer cells often have defects in one checkpoint control that makes them more vulnerable to inhibition of a second checkpoint, thereby enhancing the overall response to treatment. The G1 and G2 checkpoints are particularly crucial for the decision of a cell to arrest in the cell cycle after damage. The checkpoints are used to try to allow the repair of any damage, or to activate the apoptotic (programmed cell death) machinery. Inhibition of both G1 and G2 checkpoints in cancer cells is therefore likely to result in an induction of the death response in cancer cells. Similarly, an increasing knowledge of the molecular mechanisms that form the basis of apoptotic pathways has helped to define why cancer cells have a reduced propensity to undergo apoptosis following the activation of apoptotic inhibitory pathways or the inhibition of pro-apoptotic pathways. Therefore, the possibility to modulate these pathways is likely to result not only in the increased activity of anticancer agents, but also in an increase in their specificity. PMID- 15541959 TI - Gene expression microarray technologies in the development of new therapeutic agents. AB - We review in detail how gene expression microarray technology is benefiting all phases of the discovery, development and subsequent use of new cancer therapeutics. Global gene expression profiling is valuable in cancer classification, elucidation of biochemical pathways and the identification of potential targets for novel molecular therapeutics. We exemplify the value in tissue culture and animal models of cancer, as well as in clinical studies. The power of expression profiling alongside gene knockout or knockdown methods such as RNA interference is illustrated. The use of basal or constitutive gene expression profiling to understand and predict drug sensitivity or resistance is described. The ability of expression profiling to define detailed molecular signatures of drug action is emphasised. The approach can identify on-target and off-target effects. It can be used to identify molecular biomarkers for proof of concept studies, pharmacodynamic endpoints and prognostic markers for predicting outcome and patient selection. PMID- 15541960 TI - Interpretation of expression-profiling results obtained from different platforms and tissue sources: examples using prostate cancer data. AB - The analysis of expression signatures is a powerful tool for the classification of cancer and other tissue samples. Several protocols and platforms are available on the market, and these lead to both confirmatory and complementary results. We review the main processing techniques for cross-platform comparisons and the different tissue sources for cancer profiling. Some examples and the cross interpretation of bibliographic data related to prostate cancer are also presented. PMID- 15541961 TI - Biomarkers of ovarian tumours. AB - Ovarian cancer is one of the most aggressive gynaecological malignancies and most often the high mortality is a direct result of delays in diagnosis. The development of an ovarian cancer-specific biomarker for the early detection of disease has the capacity to improve the dismal survival rate. Currently, there are multiple investigations that are utilising both genomic and proteomic technologies to identify genes, gene products and proteins that may potentially identify diagnostic ovarian cancer biomarkers. Here, we review the studies that are involved in biomarker development for the detection of ovarian cancer. PMID- 15541962 TI - Circulating biomarkers from tumour bulk to tumour machinery: promises and pitfalls. AB - In this paper, we provide a working classification for circulating biomarkers according to their potential clinical application. We broadly divided biomarkers into four groups: (i) biomarkers of cancer risk, (ii) biomarkers of tumour-host interactions, (iii) biomarker of tumour burden, and (iv) function-related biomarkers. We hope this classification will provide a framework to which the results of future studies can be added. We also discuss the promises and pitfalls in the optional use of biomarkers in oncology. PMID- 15541963 TI - Genomic and proteomic technologies for individualisation and improvement of cancer treatment. AB - The development of microarray-based technologies for characterising tumours, both at the genomic and proteomic levels, has had a significant impact on the field of oncology. Gene expression profiling of various human tumour tissues has led to the identification of expression patterns related to disease outcome and drug resistance, as well as to the discovery of new therapeutic targets and insights into disease pathogenesis. Protein microarray technologies, such as reverse-phase protein arrays, provide the unique opportunity to profile tissues and assess the activity of signalling pathways within isolated cell populations. This technology can be used to identify patients likely to benefit from specific treatment modalities and also to monitor therapeutic response in samples obtained during and after treatment. Routine application of genomic and proteomic microarray technologies in clinical practice will require significant efforts to standardise the techniques, controls and reference standards, and analytical tools used. Extensive, independent validation using large, statistically-powered datasets will also be necessary. Inclusion of concomitant genomic and proteomic-based molecular profiling techniques into clinical trial protocols will bring us closer to the reality of patient-tailored therapy. PMID- 15541964 TI - The next challenge-adapting to change. PMID- 15541965 TI - Comparison of somatic development and status of conduit after extracardiac Fontan operation in young and older children. AB - OBJECTIVE: We set out to examine whether the extracardiac Fontan operation (ECFO) in young children is beneficial for the early postoperative course and whether it has a negative impact on the mid-term hemodynamics and growth of the children due to absent growth potential of the prosthetic conduit. Therefore we compared our medium-term experience with ECFO in children under 4 years of age to that in older children regarding the incidence of postoperative complications, somatomotoric development and conduit status. METHODS AND RESULTS: Between 11/95 and 12/02 ECFO was performed in 30 children under 4 years of age and 21 older children aged 4-13 years. There were no deaths in children under 4 years of age and two older children died. No prolonged support (>72 h) of suprarenin was required in small children compared to 4 older children. In twenty-seven children, who underwent postoperative heart catheterization no pulmonary artery or systemic vein distortion occurred. One re-operation and one transcatheter intervention were performed because of the partial conduit stenosis. During the median follow-up of 4.8 years a manifestly accelerated postoperative weight gain was observed in children operated on under 4 years of age, compared to that in older children (up to the 50 vs. 10th percentile, P<0.05). CONCLUSIONS: The ECFO could be performed in young children with low morbidity and mortality rates. In the majority of patients, implanted prosthetic grafts maintain stable form without the development of stenosis. Accelerated somatic development, especially in small children, is to be observed after completion of the Fontan circulation. PMID- 15541966 TI - Prevalence and optimal management strategy for aortic regurgitation in tetralogy of Fallot. AB - OBJECTIVE: Aortic regurgitation (AR) in the tetralogy of Fallot (TOF) is not frequent, but when present it impacts significantly on surgical management. Furthermore, the incidence of late AR development has been increasing, along with surgical interest in current practices. METHODS: Pre- and post-operative studies on 427 patients (TOF, 374; TOF/PA (TOF with pulmonary atresia), 53) who survived corrective operation were reviewed. AR (> or =mild) was detected in 28. RESULTS: Nine had AR preoperatively, while 25 (including six with preoperative AR) exhibited AR post-operatively. In the 19 who developed AR post-operatively, the aortic root diameter (AoRoD) and indexed AoRoD (%AoRoD) were 42+/-11 mm and 166+/ 36%, increased from the preoperative values of 30+/-10mm and 149+/-24%. AR-free rate at 20 years was 95.1% of all cases studied, 84.3 vs 96.5% in TOF/PA vs classic TOF (P<0.0001), and 82.2 vs 97.0% in bulboventricular VSD vs infracristal VSD (P<0.0001). Older age at repair, and bulboventricular VSD were identified as risk factors for the progression of AR. Aortic valvuloplasty (AVP; n=5) or replacement (AVR; n=4) was performed nine times in eight patients before (n=1), during (n=4), or late after TOF repair (n=4); all showed improvement of NYHA class. Survival- and reoperation-free survival curves showed no significant difference between patients with or without AR. CONCLUSIONS: After repair of TOF, careful observation for a late progression of AR is needed for the optimal timing of surgical intervention, especially in patients who repaired at higher age with a dilated aortic root or in patients with bulboventricular VSD. PMID- 15541967 TI - Modified Ross procedure using a conduit with a synthetic valve. AB - OBJECTIVE: In the Ross procedure, a homograft conduit is commonly used in place of an autotransplanted pulmonary valve. Homograft availability may be a problem and has resulted in a search for alternatives. We performed a modified Ross procedure for right ventricular outflow tract reconstruction with a synthetic valved conduit as an alternative to homograft. Our early results of valvular and right ventricular function were evaluated in patients who used a conduit with a synthetic valve. METHODS: Subjects consisted of 11 patients, who ranged in age from 5 to 22 years (12.0+/-4.9), and whose body weight ranged from 15.1 to 52.5 (34.3+/-14.4) kg. Indications for surgery were aortic stenosis (n=3), aortic stenosis and regurgitation (n=4), and aortic regurgitation (n=4). Right ventricular outflow tract reconstruction was performed using a hand-fashioned valved conduit prepared by sewing a 0.1 mm thick polytetrafluoroethylene sheet onto the luminal cavity of the 20-28 mm conduit. A conduit made with polytetrafluoroethylene was used in 8 patients, and a Dacron graft was used in 3 patients. RESULTS: There was no in-hospital or late mortality and angiocardiography at discharge revealed that all artificial valves remained active. The mean right atrial pressure and right ventricular end-diastolic pressure were not statistically different from preoperative values. The latest echocardiography (mean interval, 12.6 months) revealed that a mean pressure gradient across the synthetic valve was 11.4+/-11.1 mmHg and none of the patients had moderate or severe regurgitation. CONCLUSIONS: We demonstrated that a modified Ross procedure for right ventricular outflow tract reconstruction using a conduit with an appropriate synthetic valve is particularly effective in older children. PMID- 15541968 TI - Gene transfer of hepatocyte growth factor with prostacyclin synthase in severe pulmonary hypertension of rats. AB - OBJECTIVE: Hepatocyte growth factor (HGF) is a multi-potent growth factor, which has anti-fibrotic effects for lung injuries. In this study, we investigated whether human HGF gene transfer may attenuate the medial hypertrophy of pulmonary arteries and enhance the ameliorating effect of prostacyclin in monocrotaline (MCT)-induced pulmonary hypertension in rats. METHODS AND RESULTS: The day before MCT injection, HVJ-liposome complex with the cDNA encoding HGF gene (H group), PGIS gene (P group), and both HGF and PGIS gene (HP group) were transfected to the liver of rats as drug delivery system for the lung. Rats transfected with control vector served as controls (C group). Twenty-eight days after MCT injection, histological examination showed marked thickening of medial wall of pulmonary arteries and right ventricular hypertrophy. Percent medial wall thickness (%WT) of peripheral pulmonary arteries, pressure ratio of the right ventricle (RV) to the left ventricle (LV), and weight ratio of the RV to the LV plus septum were significantly increased in the control. Percent medial wall thickness was significantly ameliorated in H group and HP group in comparison with C group. Pressure and weight ratio of RV to LV was significantly ameliorated in P group and HP group in comparison with C group, and was significantly ameliorated in HP group than P group. CONCLUSIONS: In vivo gene transfection with HGF gene attenuated the medial hypertrophy of pulmonary arteries and enhanced the ameliorating effect of prostacyclin for pulmonary hypertension in MCT rats. Thus, gene therapy with HGF and PGIS may be a promising strategy for severe pulmonary hypertension. PMID- 15541969 TI - Tissue microarray detection of matrix metalloproteinases, in diseased tricuspid and bicuspid aortic valves with or without pathology of the ascending aorta. AB - OBJECTIVE: The degeneration of bicuspid aortic valve and its frequent association with ascending aortic pathology, point to a still unidentified genetic tissue defect with unknown mediators. Metalloproteinases (MMPs) are lytic enzymes that have been strongly implicated in aneurysm formation. The purpose of this study was to detect the presence of these enzymes in aortic valvular tissue in healthy and diseased aortic valves with or without the presence of synchronous ascending aortic pathology. METHODS: Aortic valve specimens from 26 aortic valve replacement patients as well as 4 healthy control tricuspid aortic valves were included. 10 patients had bicuspid aortic valves, and 16 had tricuspid aortic valves. Half of our patient population had a concomitant aortic procedure for aortic pathology. The study detected MMPs 1,2 and 9 as well as their Tissue inhibitors (TIMPs) 1 and 2. MMP and TIMP detection was accomplished with the construction of a tissue micro array and immunohistochemistry. CONCLUSIONS: MMP-9 expression was significantly higher in bicuspid aortic valves compared to normal valves (P<0.05). When compared to the tricuspid valve group, MMP-9 mean value was significantly higher in bicuspid valves (P<0.05). When the entire rest of the valve group (n=4+16, i.e. control and tricuspid valve groups) was compared to the bicuspid valve group, bicuspid valves had significantly higher MMP-2, and MMP-9 (P<0.01) expression. TIMP expression also changed in diseased valves, among different patient groups. This increased proteolytic presence in bicuspid aortic valves may attribute to the observed decreased elastin and collagen content, and their resultant functional failure. PMID- 15541970 TI - Surgical results for active endocarditis with prosthetic valve replacement: impact of culture-negative endocarditis on early and late outcomes. AB - OBJECTIVE: Surgical treatment of active infective endocarditis requires not only hemodynamic repair, but also special emphasis on the eradication of the infectious focus to prevent recurrence. This goal can be achieved by the combination of aggressive debridement of infective tissue and appropriate and adequate antibiotic treatment. We reviewed our experience with active endocarditis and identified factors determining early and late outcomes, particularly focusing on the factor of culture-negative endocarditis. METHODS: Sixty seven patients with clinical evidence of active endocarditis who underwent operation between 1991 and 2001 were evaluated. The aortic valve was infected in 28 (42%), the mitral valve in 23 (34%), and multiple valves in 16 (24%). Native valve endocarditis was present in 58 (87%) and prosthetic valve endocarditis in 9 (13%). Mean follow-up was 5.7 years (range, 0.2-11.5 years). RESULTS: Microorganisms were detected in 46 (69%): Staphylococcus aureus in 9 (13%), other staphylococci in 9 (13%), streptococcus species in 19 (28%), and others in 9 (28%), whereas 21 (31%) patients had culture-negative endocarditis. Operative mortality was 17.8% (12 patients). Reoperation was required in 8 (12%), while 3 late deaths (5.5% of hospital survivors) occurred. All events, including death, reoperation, periprosthetic leak, and recurrence of infection, occurred within 2 years after operation. Actuarial freedom from reoperation, late survival, and events at 5 years were 81.6, 76.4, and 68.6%, respectively. On multivariate analysis, no independent adverse predictor was detected for hospital death, whereas the following independent adverse predictors were identified: preoperative heart failure (P=0.0375), prosthetic valve endocarditis (P=0.0391) and culture-negative endocarditis (P=0.0354) for poor late survival; culture negative endocarditis (P=0.0354) and annular abscess (P=0066) for poor event-free survival. Freedom from events was similar between patients with Staphylococcus aureus infection (3-year freedom 55.6%) and culture-negative endocarditis (3-year freedom 47.6%), whereas events were significantly low in patients with streptococcus infection (3-year freedom 100%). CONCLUSIONS: In our analysis, no independent adverse predictor was detected for hospital death; however, culture negative endocarditis was identified as an independent predictor for both late survival and events after surgery. Event-free survivals were similar between staphylococcus infection and culture-negative endocarditis, and all events occurred within 2 years after operation, suggesting the necessity of close follow up during that period. PMID- 15541971 TI - Ischemic mitral valve prolapse: mechanisms and implications for valve repair. AB - OBJECTIVE: The aim of this study was to assess the mechanisms of prolapse in ischemic mitral valve regurgitation (MR) and the techniques of valve repair. METHODS: Out of 121 patients operated upon for ischemic MR, a prolapse was present in 44 patients (36.4%). The operation was performed emergently in four cases (9.1%) and electively in 40 patients (90.9%). Fifteen patients (34.1%) were operated upon within 60 days following acute myocardial infarction. RESULTS: The diagnosis of prolapse had been overlooked by echography in five cases (11.4%). A commissural area was involved as the site of prolapse in 31 cases (70.4%). The mechanism of prolapse was a papillary muscle (PM) lesion in 38 cases (86.4%) (anterior PM: n=8, posterior PM n=36) or a chordal lesion in six cases (13.6%). PM injury was elongation (n=16), or rupture (total n=1, partial n=21, incomplete n=4). The operative technique was mitral valve repair with Carpentier's techniques in 42 cases (95.5%) or replacement in two cases (4.5%). Hospital mortality was 11.4% (n=4). The mean follow-up was to 44.7+/-29.6 months. Overall survival and freedom from reoperation were 68.3+/-9.0 and 89.9+/-5.7% at 5 years, respectively. Freedom from MR equal or > grade 2 was 69.7+/-9.5% at 5 years. CONCLUSIONS: The mechanisms of ischemic mitral valve prolapse were variable and tightly linked to the PM anatomy. A reliable mitral valve repair could be achieved in most cases with acceptable mid-term results. PMID- 15541972 TI - Impact of mitral insufficiency on reoperative coronary artery surgery in ischemic cardiomyopathy patients. AB - OBJECTIVE: To determine the impact of mitral insufficiency on survival after reoperative coronary artery bypass grafting (CABG) in ischemic cardiomyopathy patients. METHODS: We retrospectively studied 891 (Initial 708, Redo 183) consecutive CABG patients (1993-2002) with ejection fraction (EF) 35% or less. PATIENT CHARACTERISTICS: mean age=67.0+/-10.5 yrs, men=77.1%, EF=26.4+/-7.4%, mean CCS=3.47+/-0.73, mean NYHA=3.50+/-0.68. There were 180 (Initial 141, Redo 39) patients with 3/4+mitral regurgitation (MR). Late survival statistics of cohorts were compiled using National Death Index. RESULTS: At a mean follow-up period of 3 years, reoperative CABG with MR (Redo/MR+) survival was 41.7+/-9.2% (n=39), which was worse than reoperative CABG without MR (Redo/MR-) survival of 71.8+/-4.1% (n=144, P=0.0003), initial CABG with MR (Initial/MR+) survival of 68.5+/-4.2% (n=141, P=0.014) and initial CABG without MR (Initial/MR-) survival of 76.2+/-2.0% (n=567, P<0.0001). By multivariate analysis, congestive heart failure (P=0.029), 3/4+MR (P=0.044) were independent predictors of Redo late mortality. In contrast, renal failure on dialysis, stroke, no angina, age >65 yrs, absence of hypercholesterolemia, EF<26% but not 3/4+MR were independent predictors of Initial late mortality. In subset analysis, adverse impact of 3/4+MR on late survival was greatest in Redo with EF<26%. The 3- and 5-yr late survival were only 44.4 and 26.8% (P=0.041). Concomitant mitral valve repair (MVrep) was performed in 100 (Initial 75, Redo 25) patients. MVrep in Initial/MR+patients achieved similar late survival as Initial/MR- patients. MVrep did not produce the same late survival benefit in Redo/MR+ patients. CONCLUSIONS: (1) Mitral insufficiency has a greater survival impact on redo than initial CABG patients with ischemic cardiomyopathy. (2) Mitral insufficiency and congestive heart failure are the primary independent predictors of late survival following redo CABG. (3) Mitral repair has less neutralizing effect on late survival in redo than initial CABG patients. PMID- 15541973 TI - Atrial fibrillation after surgical revascularization: is there any difference between on-pump and off-pump? AB - OBJECTIVE: Postoperative atrial fibrillation (AF) is still frequent complication after cardiac surgery in spite of the improvements in the surgical procedures. There is still controversy whether or not, the absence of cardiopulmonary bypass results in a lower incidence of AF. METHODS: Six hundred and seventy patients that underwent revascularization by using in situ LIMA for single vessel disease were included in this retrospective study and the patients were divided in two groups. Group I included 328 patients who underwent complete revascularization with cardiopulmonary bypass and group II consisted of 342 patients who underwent complete revascularization without cardiopulmonary bypass. Then, the incidence and predictive perioperative factors of AF in two groups were determined and compared with each other. RESULTS: There were no significant differences between two groups with respect to the preoperative demographic characteristics of the patients. The incidence of postoperative AF was determined as 16.1% after on-pump and 14.6% after off-pump revascularization. Avoiding cardiopulmonary bypass did not decrease the incidence of postoperative AF. Sex, age over 65 years, prophylactic beta-blocker usage and left ventricular dysfunction were independent predictive factors in group I (r2=0.51; P<0.001). However, only age over 65 years and prophylactic beta-blocker usage were independent predictive factors in group II (r2=0.59; P<0.01). The rates of AF in both groups were decreased by using prophylactic beta-blocker usage (P=0.05 in group I, P<0.001 in group II). CONCLUSIONS: There is no reduction of AF rate in myocardial revascularization without cardiopulmonary bypass. However, prophylactic beta-blocker usage decreases the incidence of AF after both on-pump and off-pump myocardial revascularization. PMID- 15541974 TI - Logistic versus additive EuroSCORE. A comparative assessment of the two models in an independent population sample. AB - OBJECTIVE: Validation of EuroSCORE outside the boundaries of the original database has been limited to the additive model and has occasionally shown inconsistencies. Therefore we sought to validate the logistic model and assess its predictive performance compared to the additive approach. METHODS: Twenty four hundred and twenty-six consecutive patients were prospectively assigned individual expected risks of dying calculated by the logistic and the additive EuroSCORE algorithms. Discriminating ability of the two models was tested by Receiver Operating Characteristic (ROC) curves. Calibration was assessed by the Hosmer-Lemeshow (H-T) test and further explored by additional cross-tabulations. A percent difference among the estimates was calculated and plotted across score groups. The series was then sorted by date of operation and split in halves to separately explore the potential effect of variation of performance. RESULTS: Observed mortality (5.6%) was not significantly different from the additive (5.3%) and the logistic estimates (6.9%). Both models satisfactorily discriminated outcomes (ROC areas of 0.80 and 0.79 for the logistic and the additive model, respectively). The H-T test showed that calibration was good for the logistic model (P=0.12) but turned out being inadequate for the additive model (P<0.0001). Further cross-tabulations confirmed a good correlation among observed and predicted death rates by the logistic model across all groups. The additive model, on the other hand, revealed a propensity to over-predict in medium-risk categories and under-predict in the very high-risk cases. Direct comparison of additive vs logistic estimates showed a similar behaviour demonstrating it as an intrinsic property of the additive approach. The split file analysis revealed a significantly improved outcome for patients treated in the second half of the series though the predictive performance of the two models was unaltered. CONCLUSIONS: Logistic EuroSCORE reliably predicted outcomes in our series despite the higher risk profile compared to the reference EuroSCORE sample and the observed variation in clinical performance during the study period. The additive model was less precise, exhibiting a predictive distortion which should be accounted for, particularly when employing it at the individual patient level. PMID- 15541975 TI - Long-term follow-up of total arterial myocardial revascularization using exclusively pedicle bilateral internal thoracic artery and right gastroepiploic artery. AB - OBJECTIVE: In order to reduce remote cardiac events associated with graft occlusions, arterial conduits are being increasingly utilized in coronary artery bypass grafting (CABG). While the internal thoracic artery (ITA) is the graft of choice for CABG, it is sometimes difficult or impossible to obtain a complete arterial revascularization only with ITAs in three-vessel diseases. We present our experience with total arterial myocardial revascularization with bilateral internal thoracic artery (BITA) and right gastroepiploic artery (rGEA). METHODS: From April 1994 to January 2004, 174 patients (165 male, mean age 55.9+/-7.4) underwent coronary artery bypass procedure with exclusive use of BITA and rGEA. Left ventricular ejection fraction ranged from 20 to 68% (mean 55.9+/-6.8%). Seven patients (4%) had poor ejection fraction (<0.30), 23 (13, 2%) had acute myocardial infarction, 14 (8%) had left main disease. The mean CPB time was 96.9+/-15.7 min and the mean cross clamping time was 70+/-14.2 min. The mean number of distal anastomoses was 3.3+/-0.5 per patient. RESULTS: Early mortality was 1.7%. The patients were followed for up to 9 years (mean follow-up time 6.3+/ 2.6 years). Actuarial freedom from cardiac death (including hospital death) was 97.6%, at 9 years after the operation. Actuarial freedom from angina and cardiac events at 9 years was 79, 5% and 77, 6%, respectively. No perioperative myocardial infarction occurred. None of the patients needed a redo-CABG after leaving the hospital. CONCLUSIONS: This study indicates that the myocardial revascularization in young patients with three-vessel disease using exclusively pedicle BITA and rGEA provides excellent 9-year patient survival and improvement in terms of freedom from return of angina pectoris and freedom from any cardiac related event. These results encourage the more extensive use of BITA and rGEA in selected patients with three-vessel coronary disease. PMID- 15541976 TI - Inhibition of myosin light chain kinase provides prolonged attenuation of radial artery vasospasm. AB - OBJECTIVE: Current treatments for conduit vessel vasospasm are short-acting and do not inhibit all vasospastic stimuli. This study tests the hypothesis that irreversible inactivation of myosin light chain kinase provides sustained inhibition of arterial vasoconstriction stimulated by a spectrum of vasopressors. METHODS: Canine radial artery segments were soaked for 60 min in control buffer or buffer with wortmannin, an irreversible inhibitor of myosin light chain kinase. The vessels were then thoroughly washed and contractile responses were quantified in response to a spectrum of vasopressors at 2 and 48 h after treatment. After 48 h, selected vessels were examined for morphologic changes and development of apoptosis. RESULTS: Two hours after treatment, wortmannin-soaked vessels contracted significantly less than controls in response to norepinephrine (0.19+/-0.07 g vs. 7.22+/-0.37 g, P<0.001), serotonin (0.92+/-0.35 g vs. 9.64+/ 0.67 g, P<0.001), thromboxane-mimetic U46619 (1.25+/-0.17 g vs. 10.99+/-0.50 g, P<0.001), and KCl (1.98+/-0.27 g vs.15.00+/-0.48 g, P<0.001). At 48 h, vasoconstriction remained significantly inhibited in wortmannin-treated vessels compared to control vessels in response to norepinephrine (2.36+/-0.17 vs. 6.95+/ 0.47 g, P<0.001), serotonin (4.67+/-0.39 vs. 12.42+/-0.70 g, P<0.001), U46619 (5.42+/-0.34 vs. 9.29+/-0.74 g, P=0.008), and KCl (7.49+/-0.48 vs. 13.32+/-0.60 g, P<0.001). Histology of wortmannin-treated vessels revealed no overt smooth muscle or endothelial cell damage. TUNEL staining revealed a significantly greater proportion of apoptotic smooth muscle and endothelial cells in wortmannin treated vessels as compared to controls. CONCLUSIONS: Disengaging the smooth muscle contractile apparatus by irreversibly binding myosin light chain kinase with wortmannin significantly attenuates radial artery vasoconstriction up to 48 h after brief treatment. This novel strategy may prevent vasospasm of arterial grafts from all causes for several postoperative days. PMID- 15541977 TI - 'Tamponade' following cardiac surgery: terminology and echocardiography may both mislead. AB - OBJECTIVE: Echocardiography is widely considered the gold standard for the diagnosis of tamponade. While a relatively common complication of cardiac surgery in adults, determining whether haemodynamics are compromised by a pericardial collection early post-operatively can be difficult. The aim of the current study was to determine the nature and magnitude of the diagnostic challenge posed by cardiac tamponade following cardiac surgery. We therefore examined the accuracy of echocardiography in the diagnosis of tamponade in this patient group. METHODS: From January 2000 to January 2002, 2297 adult patients underwent cardiac surgery in a tertiary referral cardiothoracic centre. A retrospective analysis of prospectively collected data, from all patients diagnosed with post-operative bleeding and/or tamponade was performed. Data included demographics, surgery, anticoagulation/anti-platelet medication, clinical/echocardiographic features of tamponade and surgical findings at re-exploration. RESULTS: The diagnosis of 'tamponade' was confirmed at re-exploration in 148 patients. When it occurred early (<72 h) following cardiac surgery trans-thoracic echocardiography failed to visualise the majority of collections (60%), necessitating trans-esophageal echocardiography. Effusions were small (160+/-17 ml) and localised (92%), showing no echocardiographic features of classical tamponade (79%). Where patients developed tamponade late (>72 h) following cardiac surgery, clinical features were atypical, effusions larger (640+/-71 ml, P<0.0001)) and global (77%). Classical echocardiographic features of tamponade were usually present (70%) and readily visualised using trans-thoracic echocardiography. CONCLUSIONS: Haemodynamically significant pericardial collections occurring early following cardiac surgery rarely cause classical clinical or echocardiographic features of tamponade. Recognition of this as a separate diagnostic entity is necessary to ensure appropriate surgical intervention is not delayed. PMID- 15541978 TI - beta2 adrenoceptor gene therapy ameliorates left ventricular dysfunction following cardiac surgery. AB - OBJECTIVE: Heart surgery is associated with impairment of the myocardial beta adrenoceptor (betaAR) system. Effective therapies for post-operative ventricular dysfunction are limited. Prolonged inotrope exposure is associated with further betaAR down-regulation. Left ventricular (LV) dysfunction and myocardial betaAR impairment were assessed following cardiopulmonary bypass (CPB) and cardioplegic arrest in a pig model. Transfer of the human beta2-adrenoceptor transgene (Adeno beta2AR) during cardioplegic arrest was then tested as a potential therapy. METHODS: Five groups of six neonatal piglets were studied. One group did not undergo surgery (Group A). Adeno-beta2AR or phosphate buffered saline (PBS) were delivered via the aortic root during cardioplegic arrest. Groups B (PBS) and C (Adeno-beta2AR) were assessed at 2 days while Groups D (PBS) and E (Adeno beta2AR) were assessed at 2 weeks from the time of surgery. An LV micromanometer was inserted under sedation to obtain pressure recordings following surgery. betaAR density was measured subsequently. RESULTS: Following cardiac surgery LV betaAR density was reduced (104+/-5.7 vs 135+/-6.1 fmol/mg membrane protein; P=0.007), and, in response to beta agonist stimulation, LV dP/dtmax was reduced (4337+/-405 vs 5328+/-194 mmHg/s; P<0.05) compared to animals which did not undergo surgery. Adeno-beta2AR therapy during cardiac surgery resulted in elevated LV betaAR density (520+/-250.9 fmol/mg) 2 days post-operatively compared to PBS (104+/-5.7 fmol/mg; P=0.002) and compared to the no surgery group (135+/ 6.1 fmol/mg; P=0.002). Elevated LV betaAR density was also present at 2 weeks (315+/-74.1 vs 119+/-7.1 fmol/mg; P=0.002). In addition, Adeno-beta2AR therapy enhanced beta agonist stimulated LV dP/dtmax (5348+/-121 vs 4337+/-405 mmHg/s; P<0.05) and heart rate (209+/-6.9 vs 173+/-11.0 bpm; P<0.05), and reduced LVEDP (2.1+/-0.4 vs 6.4+/-1.8 mmHg; P<0.05) compared to PBS treatment. Interestingly, gene delivery was cardiac-selective and beneficial effects on function persisted for 2 weeks. Moreover, beta2AR gene transfer ameliorated LV dysfunction following surgery such that there were no significant differences between non-operated controls and animals treated with Adeno-beta2AR during CPB and cardioplegic arrest. CONCLUSIONS: Reduced betaAR density and impaired LV function were present following CPB and cardioplegic arrest. Cardiac-selective beta2AR gene transfer during CPB resulted in amelioration of LV dysfunction after cardiac surgery. Such a technique may offer a new approach to post-operative ventricular support. PMID- 15541979 TI - Fulminant myocarditis in adults and children: bi-ventricular assist device for recovery. AB - OBJECTIVE: Fulminant myocarditis (FM) is uncommon and may be followed by a rapidly intractable cardiogenic shock. We report five consecutive patients with FM successfully bridged to recovery with a mechanical paracorporel biventricular assist device (BiVAD). METHODS: Five patients, four adults and one child (mean age 27+/-6 years, range, 5-36 years) underwent implantation from November 1999 to May 2003, for FM. Prior to implantation, all patients required maximal inotropic support, three of them had an intra-aortic balloon pump, the child had an extra corporel membrane oxygenation (ECMO) support previously inserted in another institution. Cardiac catheterisation showed a mean CPW of 37+/-1 mmHg, mean CVP 18+/-2 mmHg, and mean CI 1.7+/-0.1l/min. Echocardiogram showed a severe biventricular hypokinesia, without any ventricular dilatation and a mean LVEF at 12.5%. Two patients were implanted in cardiac arrest under external cardiac resuscitation. All patients underwent BiVAD implantation (MEDOS HIA-VAD). A 72 ml right paracorporel ventricle (a 23 ml in the child) was instituted between the double stage venous canula used during CPB and a pulmonary artery outflow canula. A 80 ml left paracorporel ventricle (a 25 ml in the child) was instituted between a left ventricle apical canula and an aorta outflow canula. RESULTS: There was no death. The mean duration support time was 11+/-6 days (from 7 to 21 days). Two patients experienced transitory deficiency due to a stroke. Four patients showed signs of FM on histological findings. Despite serologic examination and viral genome research on myocardial biopsies, pathogenic agents were not identified. At mean follow-up of 31+/-15 months, all the patients fully recovered with a mean LVEF=60% and no left ventricular dilatation. CONCLUSIONS: In FM with intractable cardiogenic shock, the use of a BiVAD as a bridge to recovery is a life saving approach and should be considered before multi-end organ failure. PMID- 15541980 TI - Residual fibrosis affects a long-term result of left ventricular volume reduction surgery for dilated cardiomyopathy in a rat experimental study. AB - OBJECTIVES: The aim of this study is to evaluate the relationship between left ventricular (LV) wall property and the results of LV volume reduction surgery (LVR) to treat dilated cardiomyopathy (DCM) in an experimental model. METHODS: DCM was introduced in 18 Lewis rats by autoimmunization with cardiac myosin. Among them, 12 rats underwent LVR and the rest were served as controls. They were subjected to echocardiography and cardiac catheterization for dimensional and functional measurements. The animals were sacrificed 4 weeks after surgery, and the fraction of myocardial fibrosis was calculated in 4 divided parts of the LV wall. RESULTS: Percent fibrosis varied widely from 4.7 to 45.2%. LV volume reduction surgery improved cardiac function immediately after surgery in all rats (Emax, 0.28+/-0.14 to 0.48+/-0.18 mmHg/microl; LV end-diastolic pressure, 21.0+/ 6.1 to 13.3+/-5.1 mmHg, P<0.05, respectively). Four weeks later, 6 hearts remained in good shape with smaller LV end-diastolic dimension (Dd) than baseline values (LV Dd, 9.7+/-0.6mm; fractional area change (FAC), 40.3+/-8.4%) and the other 6 had more redilation in diameter and more deterioration in function than baseline values (LV Dd, 10.9+/-0.6mm; FAC, 25.8+/-6.9%; P<0.05, respectively). Percent fibrosis in the septum differed 11.1+/-3.4 vs. 27.8+/-2.8% between the two groups (P<0.01). There was a significant correlation between the ratio of LV redilatation after surgery and percent fibrosis in the septum (r=0.951, P<0.01). CONCLUSIONS: Although the initial benefit of LVR was confirmed, the long-term result was affected by the amount of residual fibrosis. This information suggests that surgical site selection is important to achieve a good result of LV restoration surgery for DCM. PMID- 15541981 TI - Long-term outcome of lung transplantation for cystic fibrosis--Danish results. AB - OBJECTIVE: Over the last decades improvements in medical therapies have delayed the progression of lung disease in cystic fibrosis (CF). However, lung disease is still the most common cause of premature death, and lung transplantation today is the only treatment for end-stage lung disease in patients with CF. We present a retrospective review of the outcome of CF patients transplanted in Denmark since start of the national lung transplantation programme in 1992. METHODS: In a 10 year period, 47 patients with CF were listed for lung transplantation; 29 patients underwent transplantation and 18 patients died while waiting for donor organs. Eleven patients received en block double lung transplantation with direct bronchial artery revascularization and 18 patients received bilateral sequential lung transplantation. Median age at transplantation was 29 years (range 11-50). RESULTS: The perioperative mortality (< or =30 days) was 3.5% (1/29 patients). Actuarial survival of transplanted patients at 1, 3, 5 and 8 years was 89, 80, 80 and 70%, respectively. Actuarial survival of non-transplanted patients on the waiting list at 1 and 2 years was 28 and 11% (P<0.0001). Causes of death of transplanted patients were: respiratory failure on day 7 (n=1), bronchiolitis obliterans syndrome (n=2), infection (Cytomegalovirus, Aspergillus fumigatus) (n=2), bronchial anastomosis dehiscence (n=1). Pulmonary function (FEV1% predicted) improved from median 20% (range 13-31) pre-transplant to 71% (range 19 118) after 5 years (P<0.0001). Renal function (51Cr-EDTA clearance) decreased from median 97 ml/min (range 45-190) pre-transplant to 32 ml/min (range 8-84) 6 months after transplantation (P<0.001). Three patients (11%) received dialysis post-transplant of whom two underwent kidney transplantation. Immunosuppressive induction therapy with rabbit-antithymocyte-globulin compared to daclizumab resulted in fewer treatments for acute rejection within the first 3 months post transplant (P=0.05 at 5-8 weeks). Burkholderia multivorans was present in three patients pre-transplant with satisfying long-term outcome in one patient. CONCLUSIONS: Lung transplantation is a well-established life-extending treatment for patients with CF and end-stage lung disease. The operative mortality is low and CF patients have a significant early survival benefit after lung transplantation. Satisfying long-term results can be achieved in this young and severely ill group of patients. PMID- 15541982 TI - The effect of autologous fibrin sealant (Vivostat) on morbidity after pulmonary lobectomy: a prospective randomised, blinded study. AB - OBJECTIVE: Postoperative air leakage is the most frequent complication after pulmonary surgery. The development of modern surgical techniques has been influenced strongly by the need to manage air leakage effectively during pulmonary resection. This study evaluated the effect of using an autologous fibrin sealant (Vivostat) during lobectomy on morbidity following surgery. METHODS: This was a prospective, blinded, randomised clinical study. Patients undergoing lobectomy were enrolled into two groups (Vivostat or non-treatment control, 20 per group). Air leakage was measured over a 1-h period (using a mechanical suction pump) on the day of operation, and both air leakage and bleeding/exudation (drainage volume) were recorded every morning postoperatively until the chest tubes were removed. Personnel recording these parameters were blinded to the intervention received. RESULTS: Compared with the control group, mean bleeding/exudate volumes were significantly reduced in the Vivostat group (day 1,370 vs. 525 ml; total, 424 vs. 782 ml; both P<0.001), and drains were inserted for a shorter time (medians, 1 vs. 2 days, P=0.07). Significantly fewer patients had air leakage at any time in the Vivostat group (40 vs. 80%, P=0.02), and air leakage volumes were significantly lower compared with the control group (median differences: day of surgery: 0.6l/min, P=0.01; total 0.8l/min, P=0.03). Postoperative hospitalisation time was shorter in the Vivostat group than in the control group but the difference was not significant (0.5 days, P=0.12). CONCLUSIONS: Vivostat fibrin sealant significantly reduces post-surgical air leakage and drainage volumes following lobectomy in pulmonary surgery and is suitable for routine use in this procedure. PMID- 15541983 TI - Cutting lung parenchyma using the harmonic scalpel--an animal experiment. AB - OBJECTIVE: Applicability of harmonic scalpel in lung surgery was investigated using an animal model. METHODS: Air tightness, control of bleeding and features of tissue regeneration were compared in a 4-week time frame of investigation in animals in which either surgical stapler or harmonic scalpel were used for pulmonary resection. RESULTS: No significant differences between the two methods were found on a clinical and histopathological basis. CONCLUSIONS: Complete lack of granuloma formation at the resection line and in its vicinity consequently restitutio ad integrum demonstrate the advantage of the harmonic scalpel over the stapler in the circumstances investigated. Overall the vibration transmission method was shown not to be inferior to the standard methods in peripheral lung tissue resection. PMID- 15541984 TI - Bronchoplastic procedures for central carcinoid tumors: clinical experience. AB - OBJECTIVE: To evaluate the short-term and the long-term results of bronchoplastic resections (BR) performed for central carcinoid tumors (CCT). METHODS: Retrospective study of patients who underwent BR for a CCT between 1966 and 2003. RESULTS: BRs were performed in 25 patients out of 92 (27%) who were resected for CCT. Median age was 37 years. All patients were symptomatic. Preoperative bronchoscopy was diagnostic in 15 patients. The bronchoplasties performed were: 11 sleeve lobectomies, 1 sleeve segmentectomy, 8 wedge lobectomies, 4 flap lobectomies and 1 wedge segmentectomy. There were 22 typical and 3 atypical carcinoids without nodal metastasis. No major complications or mortality occurred. One patient with a typical carcinoid developed pretracheal metastatic adenopathy 19 years after resection. No recurrence or stenosis has occurred at the sites of bronchoplasty so far. Three patients died of unrelated disease. Overall the 10-, 15- and 20-year survival rates were 100, 100 and 71%. CONCLUSIONS: Bronchoplastic resections are the treatment of choice for CCT. Short and long-term results are excellent. Life-long follow-up is necessary, however, due to the possibility of late recurrence. PMID- 15541985 TI - Resection of chest wall invasion in patients with non-small cell lung cancer. AB - OBJECTIVE: The treatment of patients with non-small cell lung cancer invading the parietal pleura or chest wall is still debated. It is unsolved whether the depth of chest wall involvement or the type of resection (extrapleural or en bloc) affects long-term survival. METHODS: design, retrospective analysis; setting, Hyogo Medical Center for Adults, patients: the 97 patients who underwent surgical resection for non-small cell lung cancer involving the parietal pleura or chest wall between 1985 and 1997 were reviewed. RESULTS: Of the 97 patients, 76 had apparently complete resection, 21 had incomplete resection. The overall 5-year survival of completely resected patients was 34.2%, and that of incompletely resected patients was 14.3% (P=0.0489). In complete resection cases, the chest wall involvement was limited to the parietal pleura in 40, extended into the subpleural soft tissues in 10, and extended into the ribs in 26. The 5-year survivals were 32.5, 30.0 and 38.5%, respectively (no significant difference). The 5-year survival of completely resected patients with T3 N0 M0 disease was 44.2%, T3 N1 M0 disease 40.0%, and T3N2 M0 disease 6.2% (P=0.0019). The 5-year survival of completely resected patients with extrapleural resections was 30.0%, that of en bloc resections 38.9% (no significant difference). CONCLUSIONS: Survival of patients with lung cancer invading the chest wall or parietal pleura after resection is highly dependent on the completeness of resection and the extent of nodal involvement, but not so much on the depth of chest wall invasion or type of resection. PMID- 15541986 TI - Lung adenocarcinoma can be subtyped according to tumor dimension by computed tomography mediastinal-window setting. Additional size criteria for clinical T1 adenocarcinoma. AB - OBJECTIVE: We sought to verify that the size of the solid component, which can be evaluated using the computed tomography mediastinal-window setting, provides new criteria for CT classification of lung adenocarcinoma. METHODS: Between 1994 and September 2002, we examined 60 patients who were clinically classified with stage T1 adenocarcinoma of the lung and normal serum CEA, who underwent standard surgical procedures. Tumor maximum dimension was evaluated using two different CT imaging settings: the lung window (lDmax), and the mediastinal window (mDmax). We analyzed the relationships between prognosis or lymph node involvement and tumor dimensions. RESULTS: The mDmax was a significant (OR 1.11, P=0.02) predictive factor for lymph node metastasis. However, lDmax was not significant (P=0.83). Age, gender, lDmax, mDmax, and lymph node involvement were analyzed as predictive factors for prognosis. In univariate analysis, mDmax and lymph node involvement were significant predictive factors for prognosis (OR 1.07, P=0.01; OR 2.56, P=0.04; respectively). In multivariate analysis, mDmax was a significant predictive factor for prognosis (OR 1.06, P=0.04). We then classified the C-T1 adenocarcinoma patients into three groups according to mDmax: T1a (< or =10 mm), T1b (from 11 to 20 mm), and T1c (from 21 to 30 mm). There was a significant difference between the three groups: the disease-free 5-year survivals were 93.3, 58.1, and 32.7%, respectively (P=0.01). CONCLUSIONS: The mDmax can give additional, useful prognostic data. This finding may provide new criteria for CT classification of lung adenocarcinoma. PMID- 15541987 TI - Relationship of shuttle walk test and lung cancer surgical outcome. AB - OBJECTIVE: Surgery remains the treatment of choice in patients with potentially resectable lung carcinoma. Both the British Thoracic Society and American Chest Physician guidelines for the selection of patients with lung cancer surgery suggest the use of a shuttle walk test to predict outcome in patients with borderline lung function. The guidelines suggest that if the patient is unable to walk 250 m during a shuttle walk test, they are high risk for surgery. However, there is no published evidence to support this recommendation. Therefore, we undertook a prospective study to examine the relationship between shuttle walk test and surgical outcome in 139 patients undergoing assessment for possible lung cancer surgery. METHODS: The shuttle walk test was performed in 139 potentially resectable patients, recruited over a 2 year period, prior to surgery. One hundred and eleven patients underwent surgery. Outcome of surgery, including duration of hospital stay, complication and mortality rates was recorded. Student's t-test was used to compare the shuttle walk distance in patients with good and poor outcome from surgery. RESULTS: Mean age of patients undergoing surgery was 69 years (42-85). Mean shuttle walk distance was 395 m (145-780), with a mean oxygen desaturation of 4% (0-14) during the test. Sixty nine patients had a good surgical outcome and 34 had a poor outcome. The shuttle walk distance was not statistically different in the two outcome groups. CONCLUSION: Shuttle walk distance should not be used to predict poor surgical outcome in lung cancer patients, contrary to current recommendations. It is therefore advisable to perform a formal cardiopulmonary exercise test if at all possible. The usefulness of a shuttle walk test might be improved. It could be compared to a predicted value, as for a formal cardiopulmonary exercise test. PMID- 15541988 TI - Bilateral thoracic extraadrenal myelolipoma. AB - Myelolipoma commonly occurs in the adrenal gland and is composed of both adipose tissue and normal hematopoietic elements. Extraadrenal myelolipoma may occur in the retroperitoneum, stomach, liver, lung, and in 3% of cases even in the mediastinum. We present a 65-year-old female patient with unspecific clinical symptoms. Routine chest roentgenograms revealed bilateral widening of the posterior mediastinum. Computed tomography showed bilateral, paravertebral lesions of 4.5 and 6.5 cm in diameter, respectively. After surgical removal, bilateral thoracic myelolipoma was pathomorphologically diagnosed. The imaging differential diagnosis of bilateral solid lesions in the posterior mediastinum including lymph node metastases, lymphomas, neurogenic tumors and extramedullary hematopoietic tumors is discussed. PMID- 15541989 TI - Thirteen cases with intramural metastasis to the stomach in 1259 patients with oesophageal squamous cell carcinoma. AB - Among a total of 1259 patients with oesophageal cancer who underwent surgical resection, intramural metastasis (IMM) was verified in 93 patients (7.4%), of which IMM to the stomach (IMMS) constituted 13 patients (1.0%). In all 13 cases, the primary cancer was located in the middle or lower thoracic oesophagus and all had lymph nodes metastasis, while 12 of the 13 (92.3%) had lymphatic invasion. In our series, as the depth of invasion advanced, the number of patients with IMM or IMMS increased, although even superficial IMMS was revealed. The gross appearance of the metastatic tumours in the stomach resembled submucosal tumours. The possibility exists that metastasis via a lymphatic duct allows expansive growth in the gastric submucosae. These findings suggest that oesophageal cancer metastasizes to the stomach via a lymph duct. In conclusion, oesophageal cancer with lymphatic invasion may lead to IMM and IMMS. Therefore, careful examination for the existence of IMM, including the stomach, is required. PMID- 15541990 TI - REV (Lecompte) procedure: how much better than the Rastelli operation? PMID- 15541991 TI - Coronary flow in sinus grafts. PMID- 15541993 TI - Epidural anesthesia for coronary artery bypass surgery. PMID- 15541994 TI - Safety of off-pump coronary artery surgery: no more skepticism please! PMID- 15541997 TI - Immunologic approaches to breast cancer treatment. AB - The need for less toxic adjuvant therapies and a better understanding of the processes by which the immune system can eradicate micrometastatic disease has generated significant interest in breast cancer immunotherapy. There are many potential approaches to stimulating an immune response against a tumor, each with relative advantages and disadvantages in regards to cost, immunogenicity, and clinical applicability in treating breast cancer. This article will review the mechanisms by which the immune system can recognize and eradicate neoplastic cells and the various methods of stimulating an anti-tumor immune response. Obstacles to the clinical effectiveness of immunotherapies in breast cancer are also discussed. PMID- 15541998 TI - Nonsurgical ablation of primary breast cancer. AB - The potential for these therapies is overwhelming when one considers that as many as 90,000 patients may be candidates annually in the United States. The ideal therapy should be safe, as painless as possible, accessible, and effective in reducing local recurrence. Because of the novelty of these techniques, no therapy has met all of these goals. Even if these ideals are attained, questions remain about how to follow these patients with regard to frequency and imaging modality for local recurrence. As these technologies are refined, and they mature; it is conceivable that the future treatment of breast cancer may be less morbid than the open breast biopsies now being used for just the diagnosis of the disease. PMID- 15541999 TI - Ductal lavage for breast cancer risk assessment. AB - Ductal lavage is a minimally-invasive procedure that permits sampling of breast ductal fluids for cytopathologic analysis. The technique is performed with topical anesthesia and involves cannulation of any fluid-yielding nipple orifice with a specially-designed catheter for lavage and aspiration of the ductal system. The procedure is used for women who have clinical evidence of increased breast cancer risk; if atypia is detected, it may strengthen a woman's interest in committing to a risk-reducing strategy or a chemoprevention trial. The technology also is being used as a tool in ongoing translational research studies. PMID- 15542000 TI - Beyond lymph node staging: molecular predictors of outcome in breast cancer. AB - This article briefly reviews the exciting, yet unproven, reports of alternate methods of predicting outcomes in breast cancer and highlights new molecular methods of diagnosing, classifying, and treating this disease. The author starts by reviewing well-known factors in breast cancer with which all alternate methods of staging and predicting outcomes must be compared prospectively. PMID- 15542001 TI - Breast cancer treatment in older women. AB - This article presents the author's current approaches to the management of breast cancer in older women, with emphasis on clinical and surgical treatment of the disease in this population. There are controversies surrounding the management of breast cancer in this population regarding adjuvant therapy, radiation therapy and surgical options. We endeavor to address these issues in the article. PMID- 15542002 TI - The quality of early-stage breast cancer treatment: what can we do to improve? AB - Early-stage breast cancer is a highly curable disease with well-established protocols, including surgery, and the adjuvant modalities of regional radiation therapy, chemotherapy, and hormonal therapy. Yet, there is clear evidence that these adjuvant modalities are underused significantly. This article reviews the evidence that supports the use of efficacious local and systemic therapies in early-stage breast cancer, reasons for underuse, and interventions that have proven to be effective in ensuring the delivery of appropriate breast cancer care and suggests strategies to improve the quality of breast cancer care. PMID- 15542003 TI - Factors that affect breast cancer treatment: underserved and minority populations. AB - Breast cancer treatment in underserved populations continues to deviate from established guidelines. Significant barriers persist at the system, physician, and patient levels that ultimately may affect survival adversely. Successful strategies to reduce the disparities must be developed to improve outcomes in this population of women. PMID- 15542004 TI - Is the neuropathological 'gold standard' diagnosis dead? Implications of clinicopathological findings in an autosomal dominant neurodegenerative disorder. AB - Genetically-derived neurodegenerative disorders offer a rare opportunity to test validity of neuropathological criteria for diagnosis. Implications regarding an autosomal dominant neurodegenerative disorder (PARK 8) in which four different neuropathological diagnoses were found at autopsy are discussed. We suggest that just as there is currently no clinical 'gold standard' for Parkinson's disease, there is no pathological 'gold standard.' We conclude that in certain circumstances genetic studies may provide definitive arbitration of validity of clinical and pathological diagnostic criteria. PMID- 15542005 TI - Protective effect of caffeine against neurodegeneration in a model of Parkinson's disease in rat: behavioral and histochemical evidence. AB - Epidemiological studies have consistently demonstrated an inverse association between coffee consumption and Parkinson's disease (PD). This study was designed to investigate the beneficial effect of caffeine at a dose comparable to that of human exposure in a model of PD. For this purpose, unilateral intrastriatal 6 hydroxydopamine (6-OHDA)-lesioned rats were pretreated with caffeine (20 mg/kg; i.p.) 1 h before surgery and treated twice a day (10 mg/kg) for 1 month. Apomorphine-induced rotations and number of Nissl-stained neurons of substantia nigra pars compacta (SNC) were counted. The results demonstrated that caffeine administration for 1 month could attenuate the rotational behavior in lesioned rats and protect the neurons of SNC against 6-OHDA toxicity. PMID- 15542006 TI - The homozygote AA genotype of the alpha1-antichymotrypsin gene may confer protection against early-onset Parkinson's disease in women. AB - There has been increasing evidence suggesting that inflammatory response maybe involved in the pathogenesis of Parkinson's disease (PD). Alpha1-antichymotrypsin gene (ACT) has been regarded as a susceptibility factor for PD in the past, but the evidence remains controversial. This case-control study was designed to investigate the association of alpha1-antichymotrypsin gene (ACT) polymorphism between 210 Taiwanese patients with clinical definite sporadic PD and 260 controls, matched by age and sex. There were no differences of allelic frequency (A and T) and genotype polymorphism (AA, AT and TT) of the ACT in PD patients from the controls. However, there were significantly fewer early-onset PD (onset age younger than 60 years) or PD women carrying the homozygote AA genotype (ACT AA) than in controls (p=0.046 and 0.044, respectively). Further analysis revealed that the reduced risk of ACT-AA was particularly significant among PD women with the onset age younger than 60 years (OR=0.796, 95% CI=0.749-0.847, p<0.0001). This study shows that ACT-AA may confer a modest protection against developing early-onset PD in women. PMID- 15542007 TI - Deep brain stimulation of the subthalamic nucleus reduces antiparkinsonian medication costs. AB - This study reports a retrospective analysis of 16 patients to determine changes in medication costs associated with deep brain stimulation of the bilateral subthalamic nucleus (DBS B-STN). Antiparkinsonian medication (APMED) costs were evaluated pre- and post-operatively at 1 and 2 years, based on prescribed dosages. After treatment with DBS, patients experienced a 32% reduction in APMED costs after 1 year and a 39% reduction after 2 years. Hypothetical projections of total potential savings are presented, accounting for increasingly complex medication regimens and medication cost inflation. DBS patients may experience a significant long-term reduction in the cost of their pharmacologic treatment. PMID- 15542008 TI - Environmental factors and Parkinson's disease: a case-control study in the Tuscany region of Italy. AB - To date the aetiology of Parkinson's disease (PD) is unknown although both genetic susceptibility and environmental factors appear to play an important role in the development of the disease. Recent data have also indicated that chronic exposure to a common pesticide can reproduce the neurochemical, behavioral and neuropathological features of PD. The epidemiological studies previously carried on the prevalence of PD in population exposed to environmental factors have produced controversial results, probably because of different trial design and different analysis methods. A case-control retrospective study was conducted in a well-defined geographic area in Tuscany-Italy with the aim to identify environmental factors possibly related to PD. No significant difference between PD patients and control subjects was observed in time spent in rural or industrial residence, in well water drinking and in the exposure to herbicides and pesticides. A significant difference between patients with PD and controls was reported for cigarette smoking, controls resulting more likely cigarette smokers in comparison with PD patients. The present findings support the view of a protective effect of cigarette smoking and do not show any significant association between environmental factors and the risk of development of PD. PMID- 15542009 TI - Somatic symptom progression in idiopathic Parkinson's disease. AB - The substantia nigra (SN) pars compacta is arranged somatotopically. Symptoms of idiopathic Parkinson's disease (IPD) are caused by a lesion in this nucleus, which spreads in a stereotyped spatio-temporal pattern during the course of the disease. We investigated the order of somatic symptom progression in a group of 30 patients with IPD to determine if progression of symptoms was consistent with ordered spread of pathology through the SN pars compacta. Thirty outpatients with IPD were interviewed retrospectively about the progression of their symptoms using a semi-structured questionnaire. All the patients experienced somatic symptom progression in one of two distinct patterns, which was dependent on the location of the initial symptom. The typical pattern of progression for patients with lower limb onset was: (i) foot, (ii) leg, (iii) arm, (iv) hand, (v) face, (vi) voice and (vii) swallowing. For patients whose first symptom was in the upper limb progression to the face and lower limbs occurred roughly simultaneously or sequentially, with the typical pattern of progression: (i) hand/arm, (ii) foot/leg and face, (iii) speech and (iv) swallowing. These patterns of disease progression may reflect two distinct patterns of neuropathology within the SN pars compacta. Although the sample size was relatively small, this is the first time this phenomenon has been described in a full-length article. PMID- 15542010 TI - Validity and reliability of the PDQ-39 and the PDQ-8 in English-speaking Parkinson's disease patients in Singapore. AB - The purpose of the study was to assess the validity and reliability of the Parkinson's disease questionnaire (PDQ-39, UK English version) as well as its briefer version (the PDQ-8) among Asian patients with Parkinson's disease (PD) in Singapore. Eighty-eight patients recruited from movement disorder clinics or patient support groups completed the PDQ-39 and EQ-5D, a generic health related quality of life questionnaire previously validated for use in Singapore. PDQ-39 items showed good convergent and discriminant validity. Construct validation against the EQ-5D showed strong correlation between these scales as hypothesised (Spearman's rho: 0.53-0.71, P<0.001 for all). Factor analysis yielded a component on which all eight PDQ-39 dimensions were substantially loaded (loading range: 0.50-0.79). The PDQ-39 also demonstrated acceptable internal consistency (Cronbach's alpha: 0.59-0.94) and test-retest reliability (intra-class correlation coefficient:0.67-0.87, n=68). Six of eight PDQ-8 items showed the strongest correlation with their PDQ-39 dimensions as hypothesised. PDQ-39 and PDQ-8 summary indexes were highly correlated (Pearson's r=0.96, ICC=0.95), and both were strongly correlated with the EQ-5D utility index and EQ-5D visual analogue scale (P<0.001). We conclude that the PDQ-39 and PDQ-8 are valid and reliable disease-specific HRQoL instruments for PD in Singapore. PMID- 15542011 TI - Dynamic estimation of hand position is abnormal in Parkinson's disease. AB - Parkinson's disease (PD) is widely viewed as a disorder of central motor control. However, recent studies suggest that disordered kinesthetic processing may also contribute to bradykinesia and hypometria in PD. To examine the hypothesis that abnormal kinesthesia in PD would result in impaired hand motion estimation used for motor control, we tested PD patients, elderly people, and young adults in an active, multi-joint kinesthetic-to-visual matching task. To minimize initial localization errors, visual information about the starting position was always available. The participants performed center-out drawing movements to visual targets in the absence of visual feedback of hand/pen motion at their preferred speed. Movement time (MT), end-point position error (EPE), and initial directional error (IDE) were measured. No detrimental effects of aging were observed; however, the PD group showed prolonged MTs and higher EPE scores as compared to the elderly and young groups. Principal component analysis of the end point error distributions showed that the PD patients had larger variability in both the extent and direction axes. These results suggest that PD patients have abnormal proprioception and deficits in the central processing and integration of kinesthetic signals, resulting in the incorrect assembly of multiple sensorimotor inputs into a motor plan. It is hypothesized that altered kinesthesia in PD causes improper estimation of hand motion used for motor control due to the degraded maintenance of a dynamic internal hand representation. PMID- 15542012 TI - Short-term and practice effects of metronome pacing in Parkinson's disease patients with gait freezing while in the 'on' state: randomized single blind evaluation. AB - OBJECTIVE: In a randomized single blind parallel study, we tested the efficacy of an auditory metronome on walking speed and freezing in Parkinson's disease (PD) patients with freezing gait impairment during their 'on' function. BACKGROUND: No pharmacological treatment is effective in managing 'on' freezing in PD. Like visual cues that can help overcome freezing, rhythmic auditory pacing may provide cues that help normalize walking pace and overcome freezing. DESIGN/METHODS: Non demented PD patients with freezing during their 'on' state walked under two conditions, in randomized order: unassisted walking and walking with the use of an audiocassette with a metronome recording. The walking trials were randomized and gait variables were rated from videotapes by a blinded evaluator. Outcome measures were total walking time (total trial time-total freezing time), which was considered the time over a course of specified length, freezing time, average freeze duration and number of freezes. All outcomes were averaged across trials for each person and then compared across conditions using Signed Rank tests. RESULTS: Twelve non-demented PD patients with a mean age of 65.8 +/- 11.2 years, and mean PD duration of 12.4 +/- 7.3 years were included. The use of the metronome slowed ambulation and increased the total walking time (P < 0.0005) only during the first visit, without affecting any freezing variable. In the nine patients who took the metronome recording home and used it daily for 1 week while walking, freezing remained unimproved. CONCLUSIONS: Though advocated in prior publications as a walking aid for PD patients, auditory metronome pacing slows walking and is not a beneficial intervention for freezing during their 'on' periods. PMID- 15542014 TI - Gene name identification and normalization using a model organism database. AB - Biology has now become an information science, and researchers are increasingly dependent on expert-curated biological databases to organize the findings from the published literature. We report here on a series of experiments related to the application of natural language processing to aid in the curation process for FlyBase. We focused on listing the normalized form of genes and gene products discussed in an article. We broke this into two steps: gene mention tagging in text, followed by normalization of gene names. For gene mention tagging, we adopted a statistical approach. To provide training data, we were able to reverse engineer the gene lists from the associated articles and abstracts, to generate text labeled (imperfectly) with gene mentions. We then evaluated the quality of the noisy training data (precision of 78%, recall 88%) and the quality of the HMM tagger output trained on this noisy data (precision 78%, recall 71%). In order to generate normalized gene lists, we explored two approaches. First, we explored simple pattern matching based on synonym lists to obtain a high recall/low precision system (recall 95%, precision 2%). Using a series of filters, we were able to improve precision to 50% with a recall of 72% (balanced F-measure of 0.59). Our second approach combined the HMM gene mention tagger with various filters to remove ambiguous mentions; this approach achieved an F-measure of 0.72 (precision 88%, recall 61%). These experiments indicate that the lexical resources provided by FlyBase are complete enough to achieve high recall on the gene list task, and that normalization requires accurate disambiguation; different strategies for tagging and normalization trade off recall for precision. PMID- 15542015 TI - Enhancing HMM-based biomedical named entity recognition by studying special phenomena. AB - The purpose of this research is to enhance an HMM-based named entity recognizer in the biomedical domain. First, we analyze the characteristics of biomedical named entities. Then, we propose a rich set of features, including orthographic, morphological, part-of-speech, and semantic trigger features. All these features are integrated via a Hidden Markov Model with back-off modeling. Furthermore, we propose a method for biomedical abbreviation recognition and two methods for cascaded named entity recognition. Evaluation on the GENIA V3.02 and V1.1 shows that our system achieves 66.5 and 62.5 F-measure, respectively, and outperforms the previous best published system by 8.1 F-measure on the same experimental setting. The major contribution of this paper lies in its rich feature set specially designed for biomedical domain and the effective methods for abbreviation and cascaded named entity recognition. To our best knowledge, our system is the first one that copes with the cascaded phenomena. PMID- 15542016 TI - Comparison of character-level and part of speech features for name recognition in biomedical texts. AB - The immense volume of data which is now available from experiments in molecular biology has led to an explosion in reported results most of which are available only in unstructured text format. For this reason there has been great interest in the task of text mining to aid in fact extraction, document screening, citation analysis, and linkage with large gene and gene-product databases. In particular there has been an intensive investigation into the named entity (NE) task as a core technology in all of these tasks which has been driven by the availability of high volume training sets such as the GENIA v3.02 corpus. Despite such large training sets accuracy for biology NE has proven to be consistently far below the high levels of performance in the news domain where F scores above 90 are commonly reported which can be considered near to human performance. We argue that it is crucial that more rigorous analysis of the factors that contribute to the model's performance be applied to discover where the underlying limitations are and what our future research direction should be. Our investigation in this paper reports on variations of two widely used feature types, part of speech (POS) tags and character-level orthographic features, and makes a comparison of how these variations influence performance. We base our experiments on a proven state-of-the-art model, support vector machines using a high quality subset of 100 annotated MEDLINE abstracts. Experiments reveal that the best performing features are orthographic features with F score of 72.6. Although the Brill tagger trained in-domain on the GENIA v3.02p POS corpus gives the best overall performance of any POS tagger, at an F score of 68.6, this is still significantly below the orthographic features. In combination these two features types appear to interfere with each other and degrade performance slightly to an F score of 72.3. PMID- 15542017 TI - Biomedical named entity recognition using two-phase model based on SVMs. AB - Named entity (NE) recognition has become one of the most fundamental tasks in biomedical knowledge acquisition. In this paper, we present a two-phase named entity recognizer based on SVMs, which consists of a boundary identification phase and a semantic classification phase of named entities. When adapting SVMs to named entity recognition, the multi-class problem and the unbalanced class distribution problem become very serious in terms of training cost and performance. We try to solve these problems by separating the NE recognition task into two subtasks, where we use appropriate SVM classifiers and relevant features for each subtask. In addition, by employing a hierarchical classification method based on ontology, we effectively solve the multi-class problem concerning semantic classification. The experimental results on the GENIA corpus show that the proposed method is effective not only in reducing computational cost but also in improving performance. The F-score (beta=1) for the boundary identification is 74.8 and the F-score for the semantic classification is 66.7. PMID- 15542018 TI - Enhancing performance of protein and gene name recognizers with filtering and integration strategies. AB - Named entity (NE) recognition is a fundamental task in biological relationship mining. This paper considers protein/gene collocates extracted from biological corpora as restrictions to enhance the precision rate of protein/gene name recognition. In addition, we integrate the results of multiple NE recognizers to improve the recall rates. Yapex and KeX, and ABGene and Idgene are taken as examples of protein and gene name recognizers, respectively. The precision of Yapex increases from 70.90 to 85.84% at the low expense of the recall rate (i.e., it only decreases 2.44%) when collocates are incorporated. When both filtering and integration strategies are employed together, the Yapex-based integration with KeX shows good performance, i.e., the F-score increases by 7.83% compared to the pure Yapex method. The results of gene recognition show the same tendency. The ABGene-based integration with Idgene shows a 10.18% F-score increase compared to the pure ABGene method. These successful methodologies can be easily extended to other name finders in biological documents. PMID- 15542019 TI - Improving the performance of dictionary-based approaches in protein name recognition. AB - Dictionary-based protein name recognition is often a first step in extracting information from biomedical documents because it can provide ID information on recognized terms. However, dictionary-based approaches present two fundamental difficulties: (1) false recognition mainly caused by short names; (2) low recall due to spelling variations. In this paper, we tackle the former problem using machine learning to filter out false positives and present two alternative methods for alleviating the latter problem of spelling variations. The first is achieved by using approximate string searching, and the second by expanding the dictionary with a probabilistic variant generator, which we propose in this paper. Experimental results using the GENIA corpus revealed that filtering using a naive Bayes classifier greatly improved precision with only a slight loss of recall, resulting in 10.8% improvement in F-measure, and dictionary expansion with the variant generator gave further 1.6% improvement and achieved an F measure of 66.6%. PMID- 15542020 TI - Use of morphological analysis in protein name recognition. AB - Protein name recognition aims to detect each and every protein names appearing in a PubMed abstract. The task is not simple, as the graphic word boundary (space separator) assumed in conventional preprocessing does not necessarily coincide with the protein name boundary. Such boundary disagreement caused by tokenization ambiguity has usually been ignored in conventional preprocessing of general English. In this paper, we argue that boundary disagreement poses serious limitations in biomedical English text processing, not to mention protein name recognition. Our key idea for dealing with the boundary disagreement is to apply techniques used in Japanese morphological analysis where there are no word boundaries. Having evaluated the proposed method with GENIA corpus 3.02, we obtain F-measure of 69.01 on a strict criterion and 79.32 on a relaxed criterion. The result is comparable to other published work in protein name recognition, without resorting to manually prepared ad hoc feature engineering. Further, compared to the conventional preprocessing, the use of morphological analysis as preprocessing improves the performance of protein name recognition and reduces the execution time. PMID- 15542021 TI - Using automatically learnt verb selectional preferences for classification of biomedical terms. AB - In this paper, we present an approach to term classification based on verb selectional patterns (VSPs), where such a pattern is defined as a set of semantic classes that could be used in combination with a given domain-specific verb. VSPs have been automatically learnt based on the information found in a corpus and an ontology in the biomedical domain. Prior to the learning phase, the corpus is terminologically processed: term recognition is performed by both looking up the dictionary of terms listed in the ontology and applying the C/NC-value method for on-the-fly term extraction. Subsequently, domain-specific verbs are automatically identified in the corpus based on the frequency of occurrence and the frequency of their co-occurrence with terms. VSPs are then learnt automatically for these verbs. Two machine learning approaches are presented. The first approach has been implemented as an iterative generalisation procedure based on a partial order relation induced by the domain-specific ontology. The second approach exploits the idea of genetic algorithms. Once the VSPs are acquired, they can be used to classify newly recognised terms co-occurring with domain-specific verbs. Given a term, the most frequently co-occurring domain-specific verb is selected. Its VSP is used to constrain the search space by focusing on potential classes of the given term. A nearest-neighbour approach is then applied to select a class from the constrained space of candidate classes. The most similar candidate class is predicted for the given term. The similarity measure used for this purpose combines contextual, lexical, and syntactic properties of terms. PMID- 15542022 TI - Using name-internal and contextual features to classify biological terms. AB - There has been considerable work done recently in recognizing named entities in biomedical text. In this paper, we investigate the named entity classification task, an integral part of the named entity extraction task. We focus on the different sources of information that can be utilized for classification, and note the extent to which they are effective in classification. To classify a name, we consider features that appear within the name as well as nearby phrases. We also develop a new strategy based on the context of occurrence and show that they improve the performance of the classification system. We show how our work relates to previous works on named entity classification in the biological domain as well as to those in generic domains. The experiments were conducted on the GENIA corpus Ver. 3.0 developed at University of Tokyo. We achieve f value of 86 in 10-fold cross validation evaluation on this corpus. PMID- 15542023 TI - Term identification in the biomedical literature. AB - Sophisticated information technologies are needed for effective data acquisition and integration from a growing body of the biomedical literature. Successful term identification is key to getting access to the stored literature information, as it is the terms (and their relationships) that convey knowledge across scientific articles. Due to the complexities of a dynamically changing biomedical terminology, term identification has been recognized as the current bottleneck in text mining, and--as a consequence--has become an important research topic both in natural language processing and biomedical communities. This article overviews state-of-the-art approaches in term identification. The process of identifying terms is analysed through three steps: term recognition, term classification, and term mapping. For each step, main approaches and general trends, along with the major problems, are discussed. By assessing previous work in context of the overall term identification process, the review also tries to delineate needs for future work in the field. PMID- 15542024 TI - Periosteum: biology, regulation, and response to osteoporosis therapies. AB - Periosteum contains osteogenic cells that regulate the outer shape of bone and work in coordination with inner cortical endosteum to regulate cortical thickness and the size and position of a bone in space. Induction of periosteal expansion, especially at sites such as the lumbar spine and femoral neck, reduces fracture risk by modifying bone dimensions to increase bone strength. The cell and molecular mechanisms that selectively and specifically activate periosteal expansion, as well as the mechanisms by which osteoporosis drugs regulate periosteum, remain poorly understood. We speculate that an alternate strategy to protect human bones from fracture may be through targeting of the periosteum, either using current or novel agents. In this review, we highlight current concepts of periosteal cell biology, including their apparent differences from endosteal osteogenic cells, discuss the limited data regarding how the periosteal surface is regulated by currently approved osteoporosis drugs, and suggest one potential means through which targeting periosteum may be achieved. Improving our understanding of mechanisms controlling periosteal expansion will likely provide insights necessary to enhance current and develop novel interventions to further reduce the risk of osteoporotic fractures. PMID- 15542025 TI - Influence of the degradation of the organic matrix on the microscopic fracture behavior of trabecular bone. AB - In recent years, the important role of the organic matrix for the mechanical properties of bone has become increasingly apparent. It is therefore of great interest to understand the interactions between the organic and inorganic constituents of bone and learn the mechanisms by which the organic matrix contributes to the remarkable properties of this complex biomaterial. In this paper, we present a multifaceted view of the changes of bone's properties due to heat-induced degradation of the organic matrix. We compare the microscopic fracture behavior (scanning electron microscopy; SEM), the topography of the surfaces (atomic force microscopy; AFM), the condition of bone constituents [X ray diffraction (XRD), thermogravimetric analysis (TGA), and gel electrophoresis], and the macromechanical properties of healthy bovine trabecular bone with trabecular bone that has a heat-degraded organic matrix. We show that heat treatment changes the microfracture behavior of trabecular bone. The primary failure mode of untreated trabecular bone is fibril-guided delamination, with mineralized collagen filaments bridging the gap of the microcrack. In contrast, bone that has been baked at 200 degrees C fractures nondirectionally like a brittle material, with no fibers spanning the microcracks. Finally, bone that has been boiled for 2 h in PBS solution fractures by delamination with many small filaments spanning the microcracks, so that the edges of the microcracks become difficult to distinguish. Of the methods we used, baking most effectively weakens the mechanical strength of bone, creating the most brittle material. Boiled bone is stronger than baked bone, but weaker than untreated bone. Boiled bone is more elastic than untreated bone, which is in turn more elastic than baked bone. These studies clearly emphasize the importance of the organic matrix in affecting the fracture mechanics of bone. PMID- 15542026 TI - Three children with lower limb fractures and a mineralization defect: a novel bone fragility disorder? AB - In this report, we describe three unrelated children with an apparently novel bone fragility disorder that is associated with an idiopathic mineralization defect. Recurrent lower limb fractures started with weight bearing. The patients had none of the phenotypic, radiological, or histomorphometric features classically associated with known bone fragility disorders such as osteogenesis imperfecta (OI), idiopathic juvenile osteoporosis (IJO), or mild autosomal dominant osteopetrosis. Radiologically, there was increased metaphyseal trabeculation, normal to increased cortical thickness, and no evidence of rickets or osteomalacia. Areal and volumetric bone mineral density (BMD) of the lumbar spine did not show any major alteration. Peripheral quantitative computed tomography of the radius showed elevated cortical thickness and total and trabecular volumetric bone mineral density in one patient. Qualitative histology of iliac bone biopsy specimens showed a paucity of the birefringent pattern of normal lamellar bone. Quantitative histomorphometric analysis demonstrated osteomalacia with a prolonged mineralization lag time in the presence of a decreased mineral apposition rate. There was no biochemical evidence of abnormal calcium or phosphate metabolism. Type I collagen mutation analysis was negative. We conclude that this is a bone fragility disorder of moderate severity that tends to cause fractures in the lower extremities and is associated with the accumulation of osteoid due to an intrinsic mineralization defect. The pathogenetic basis for this disorder remains to be elucidated. PMID- 15542027 TI - A family history of fracture and fracture risk: a meta-analysis. AB - The aims of the present study were to determine whether a parental history of any fracture or hip fracture specifically are significant risk factors for future fracture in an international setting, and to explore the effects of age, sex and bone mineral density (BMD) on this risk. We studied 34,928 men and women from seven prospectively studied cohorts followed for 134,374 person-years. The cohorts comprised the EPOS/EVOS study, CaMos, the Rotterdam Study, DOES and cohorts at Sheffield, Rochester and Gothenburg. The effect of family history of osteoporotic fracture or of hip fracture in first-degree relatives, BMD and age on all clinical fracture, osteoporotic fracture and hip fracture risk alone was examined using Poisson regression in each cohort and for each sex. The results of the different studies were merged from the weighted beta coefficients. A parental history of fracture was associated with a modest but significantly increased risk of any fracture, osteoporotic fracture and hip fracture in men and women combined. The risk ratio (RR) for any fracture was 1.17 (95% CI=1.07-1.28), for any osteoporotic fracture was 1.18 (95% CI=1.06-1.31), and for hip fracture was 1.49 (95% CI=1.17-1.89). The risk ratio was higher at younger ages but not significantly so. No significant difference in risk was seen between men and women with a parental history for any fracture (RR=1.17 and 1.17, respectively) or for an osteoporotic fracture (RR=1.17 and 1.18, respectively). For hip fracture, the risk ratios were somewhat higher, but not significantly higher, in men than in women (RR=2.02 and 1.38, respectively). A family history of hip fracture in parents was associated with a significant risk both of all osteoporotic fracture (RR 1.54; 95CI=1.25-1.88) and of hip fracture (RR=2.27; 95% CI=1.47-3.49). The risk was not significantly changed when BMD was added to the model. We conclude that a parental history of fracture (particularly a family history of hip fracture) confers an increased risk of fracture that is independent of BMD. Its identification on an international basis supports the use of this risk factor in case-finding strategies. PMID- 15542028 TI - Intravenous pamidronate treatment of children under 36 months of age with osteogenesis imperfecta. AB - INTRODUCTION: Bone mineral density (BMD) and fracture rates in children with osteogenesis imperfecta (OI) have been shown to improve with bisphosphonate therapy. There are limited data available on the efficacy of this therapy in children with OI under the age of 3 years. To examine this, we instituted a prospective clinical trial of intravenous bisphosphonate to study safety, feasibility, and efficacy of this therapy. MATERIALS AND METHODS: Nine infants and young children with osteogenesis imperfecta (age range 1-35 months) were treated with intravenous pamidronate. Six had type II OI, two had type I, and one had type IV. Pamidronate was administered in cycles of 3 consecutive days. The total duration of therapy ranged from 11 to 29 months (mean 17 months). RESULTS: During treatment, the mean annualized percent change in total body areal BMD was 25% (range 11-40%). Pamidronate therapy resulted in sustained and significant decreases in serum calcium and bone-specific alkaline phosphatase and in urine calcium/creatinine and NTX/creatinine. Fracture rate in the group decreased from 80 fractures in 111 months before treatment to 25 fractures in 152 months after treatment (P<0.01). Linear growth and weight gain were maintained. Other than fevers in several infants following the initial dose of intravenous bisphosphonate no adverse effects of therapy were noted. CONCLUSIONS: Our data support that intravenous pamidronate therapy is safe, increases BMD, and reduces fracture rates in very young children with OI. Currently, it would seem to be the best available treatment for these children. PMID- 15542029 TI - Congenic mice with low serum IGF-I have increased body fat, reduced bone mineral density, and an altered osteoblast differentiation program. AB - Targeted gene studies have demonstrated the importance of insulin-like growth factor-I (IGF-I) for osteoblast (OB) differentiation and the acquisition of peak bone mineral density (BMD). The skeletal response to allelic differences in IGF-I expression can also be measured in vivo, using congenic mice. We created a congenic strain with reduced (approximately 20%) circulating IGF-I (C3H.B6-6T [6T]) by backcrossing a small genomic region (30 cM) of Chromosome 6 (Chr6) from C3H/HeJ (C3H) onto a C57Bl/6J (B6) background. 6T female mice have lower serum IGF-I (P<0.001 vs. B6) but similar growth hormone (GH) and serum IGF binding protein (IGFBP) concentrations as B6. At 16 weeks of age, congenics have greater body fat (P<0.02 vs. B6) despite less total body weight, and exhibit smaller femoral cross-sectional size (P=0.001), reduced cortical thickness (P<0.001) and lower trabecular BV/TV (P<0.05) than B6. 6T mice also have suppressed serum leptin (P<0.01), but compared to B6 have similar markers of bone resorption (i.e., urine CTx and serum TRAP 5B). At 8 weeks of age, skeletal IGF-I mRNA from long bones was reduced by 40% (P<0.05) as were liver mRNA transcripts (i.e., 50%, P<0.01). Osteoblast progenitors from the bone marrow of 6T mice formed less colony forming unit fibroblasts by crystal violet staining than B6 (P<0.007) and had significantly reduced alkaline phosphatase-positive colonies than B6(P<0.0001). In addition, staining of bone marrow with oil red O revealed greater numbers of adipocytes in 6T than B6. Several candidate genes in the Chr6 QTL were excluded by lack of strain-related expression differences in bone, but genes positively regulating adipocyte differentiation including Alox 5 and PPAR gamma require further study as either "pathway" or candidate genes. In summary, allelic differences in a QTL on Chr6 result in altered IGF-I gene expression, changes in OB lineage allocation, and reduced peak bone mass. Congenic mice are useful models not only for mapping genes related to bone mass but also for elucidating the biology underlying various skeletal phenotypes associated with more subtle manipulation of the mouse genome. PMID- 15542030 TI - The deficiency of immunoregulatory receptor PD-1 causes mild osteopetrosis. AB - Recently, the involvement of immune responses in metabolic bone disease and/or local bone destruction has received much attention. Cytotoxic T lymphocyte associated antigen 4 (CTLA-4), a member of the immunoglobulin (Ig) superfamily, negatively regulates T cell activation. The deficiency of CTLA-4 induces profound osteopenia with an increase in osteoclastogenesis, suggesting the important role of activated T cells in osteoclastogenesis. Programmed death-1 (PD-1) is the newly identified immunoregulatory receptor, which also belongs to the Ig superfamily. Both CTLA-4 and PD-1 are induced on activated T cells, however, there are no reports linking PD-1 with osteoclasts. In the present study, we have examined the bone phenotype in PD-1-deficient mice PD-1-/- and the role of PD-1 in osteoclastogenesis and osteoclast function. Both trabecular and cortical bone mineral densities of tibia were significantly increased, as observed in peripheral quantitative computed tomography (pQCT), at 12 weeks of age in PD-1-/- mice. Histomorphometric analysis of the PD-1-/- mice and the age-matched controls at 12 weeks of age showed a 2-fold increase in bone volume (BV/TV) with a 55% decrease in osteoclast number (N.Oc/BS). Bone formation indices were similar in both groups. The number of soluble receptor activator of nuclear factor kappaB ligand (sRANKL)-induced osteoclast-like cells (OCLs) derived from the PD-1 deficient splenocytes was significantly decreased (by 25%). On the other hand, PD 1 deficiency did not affect the bone-resorbing activity of mature osteoclasts. Our results suggest that PD-1 deficiency reduces osteoclastogenesis resulting in an osteopetrotic phenotype. Identical members of the Ig superfamily, CTLA-4 and PD-1, which negatively regulate immune responses, may differentially affect osteoclastogenesis and bone remodeling. PMID- 15542031 TI - Developmental failure of phalanges in the absence of growth/differentiation factor 5. AB - Growth/differentiation factor 5 (GDF5) is a member of the bone morphogenetic protein (BMP) family, which has been implicated in several skeletogenic events including cartilage and bone formation. To study the role of GDF5, we analyzed digit development in brachypodism (bp) mice, which carry functional null mutations of the Gdf5 gene and exhibit a reduction in the length of digit bones and loss of the middle phalanges. In situ detection of apoptosis and whole-mount detection of cell death showed abnormal apoptosis in the developing phalanges of bp mice. In situ hybridization in bp mice showed overexpression of Gdf5 mRNA in the developing phalanges where apoptotic cells were increased. In addition, bp mice exhibited excessive apoptosis in the interdigital regions. The condensed mesenchymal cells were progressively decreased in the developing phalanges and failed to form cartilage models of the middle phalanges. These findings show that excessive apoptosis in the absence of GDF5 results in developmental failure of the phalanges. We conclude that GDF5 is essential for maintenance and growth of the developing phalanges. PMID- 15542032 TI - Site-specific localization of two distinct phosphatases along the osteoblast plasma membrane: tissue non-specific alkaline phosphatase and plasma membrane calcium ATPase. AB - In osteoblasts, alkaline phosphatase has been reported to be restricted to the basolateral domains. In recent studies, we have demonstrated phosphatase activities different from those of tissue non-specific alkaline phosphatase (TNSALP) along the osteoidal aspect of osteoblast membrane at alkaline and neutral pH on undecalcified freshly frozen sections of rat bones. In the present study, we sought to further characterize and define the nature of membrane associated phosphatases along the osteoidal aspect of osteoblasts. Histochemical properties of the enzymes and their localization in vivo were examined in long bones of normal Wistar rats and TNSALP null mutant mice and their wild type littermates. Molecular profiles of the enzymes in the osteoblast extracts were also examined. The enzymatic activity of the phosphatase along the osteoidal surface of osteoblasts proved to be activated by both Mg2+ and Ca2+. Unlike TNSALP, the activity was inhibited by vanadate but resistant to levamisole, implicating a similarity between this enzyme and plasma membrane Ca2+ transport ATPase (PMCA). Immunohistochemistry showed that PMCA immunoreactions were restricted to the osteoidal domain of the plasma membrane. Native-PAGE analysis of osteoblast extracts suggested the presence of two phosphatases corresponding, respectively, to TNSALP and PMCA. Western blot analysis after SDS-PAGE of osteoblast extracts confirmed the existence of PMCA (140 kDa) and TNSALP (80 kDa). Gel-chemical analysis of the osteoblast extract from TNSALP null mutant mice depicted phosphatase activity, which was resistant to levamisole. These data suggest the presence of a phosphatase different from TNSALP, most plausibly PMCA, on the osteoidal surface of osteoblasts. PMID- 15542033 TI - Mapping quantitative trait loci that influence blood levels of alkaline phosphatase in MRL/MpJ and SJL/J mice. AB - To examine the hypothesis that serum alkaline phosphatase (ALP) levels have a heritable component, we analyzed blood from two inbred strains of mice, MRL/MpJ and SJL, which exhibit 90% difference in total serum ALP activity (268+/-26 vs. 140+/-15 U/l, respectively, P<0.001). A genome-wide scan was carried out using 137 polymorphic markers in 518 F2 female mice. Serum ALP activity in the F2 progeny showed a normal distribution with an estimated heritability of 56%. Genome-wide scan for cosegregation of genetic marker data with serum ALP activity revealed three major quantitative trait loci (QTL), one each on chromosomes 2 (LOD score 3.8), chromosome 6 (LOD score 12.0), and chromosome 14 (LOD score 3.7). In addition, there was one suggestive QTL on chromosome 2 (LOD score of 3.3). In aggregate, these QTLs explain 22.5% of variance in serum ALP between these two strains. Serum ALP showed a moderate but significant correlation with body weight adjusted total body bone mineral density (r=0.12, P=0.0108) and periosteal circumference at midshaft tibia (r=0.15, P=0.0006) in F2 mice. The chromosome 6 locus harboring the major serum ALP QTL also contains a major BMD and bone size QTL, identified earlier, between these two strains of mice; in addition, this QTL is also close to the locus that regulates IGF-I levels (LOD score 8-9) in C3HB6 F2 mice. These common QTLs indicate that the observed difference in ALP and BMD or bone size may be regulated by same loci (or genes). Accordingly, the osteoblast cells isolated from femur and tibia of MRL mice showed a significantly higher number of ALP +ve cells/colony and two- to threefold higher ALP activity (P<0.001) as compared to the cells isolated from SJL mice, thus suggesting that differences in serum ALP between MRL and SJL reflect difference in ALP expression from osteoblasts from these strains of mice. These data suggest that serum ALP levels are genetically determined and correlate with cellular mechanisms that differentiate BMD accrual in these two strains of mice. The findings that ALP and BMD traits share the same loci on chromosome 6 suggest a role for genetic determinants of bone formation in overall BMD accretion. PMID- 15542034 TI - Osteocyte density in woven bone. AB - Woven bone forms rapidly during tissue growth, following injury and in response to certain anabolic stimuli. Functional differences between woven and lamellar bone may be due, in part, to differences in osteocyte density (cells per unit tissue). Woven bone has been estimated to contain four to eight times more osteocytes than lamellar bone, although primary data to support this assertion are limited. Given recent findings implicating osteocytes as regulators of bone remodeling, bone formation and bone volume, such large differences in osteocyte density between woven and lamellar bone may have important consequences. In this study, we compared the density of osteocyte lacunae (lacunae/mm(2) tissue) in rat lamellar bone with that in woven bone formed under several different circumstances. We found that the lacunar density of lamellar cortical bone in the rat (834+/-83 cells/mm2, mean+/-SD) did not differ significantly from that of periosteal woven bone formed via intramembranous osteogenesis, either in response to mechanical loading (921+/-204 cells/mm2) or in the periosteal buttressing region of the fracture callus (1138+/-168 cells/mm2). In contrast, lacunar density of endochondrally derived woven bone in the center (gap) region of fracture callus was nearly 100% greater (1875+/-270 cells/mm2) than in lamellar cortical bone while lacunar density of primary spongiosa of the growth plate was 40% greater (1674+/-228 cells/mm2) than that in lamellar cancellous bone (1189+/ 164). These findings demonstrate that lacunar density in woven bone varies depending on skeletal site and developmental history and appears to be elevated in endochondrally derived woven bone adjacent to marrow space. Given the considerable evidence supporting osteocytes as local initiators of bone remodeling, we suggest that woven bone with increased lacunar density may undergo remodeling at an accelerated rate. PMID- 15542035 TI - Accelerated bone formation and increased osteoblast number contribute to the abnormal tooth germ development in parathyroid hormone-related protein knockout mice. AB - Our previous study showed that tooth germs at late embryonic stage [later than embryonic day 17.5 (E17.5)] and neonatal homozygous parathyroid hormone-related protein (PTHrP)-knockout mice are compressed or penetrated by the surrounding alveolar bone tissue. In vivo and in vitro studies have shown that the development of the tooth germ proper is not disturbed, but insufficient alveolar bone resorption, due to the decreased number and hypofunction of osteoclasts, is the main cause of this abnormality. In addition to the insufficient alveolar bone resorption, progressive bone formation toward tooth germs was observed in homozygous mice, suggesting that accelerated bone formation also contributes to this abnormality. To further investigate this, homozygous mice at E14.0 and E15.5, when alveolar bone is forming, were used for histochemical and bone histomorphometric analyses. In contrast to the late embryonic stage, the alveolar bone did not yet compress developing tooth germs in homozygous mice on E14.0, but a larger amount of bone tissue was seen compared to wild-type littermates. Histomorphometric analysis of bone at E14.0 revealed that the osteoblast numbers and surfaces in the mandibles and in the bone collar of femora of homozygous mice were significantly higher than those of wild-type mice. However, unlike our previous study showing the osteoclast surface on E18.5 in homozygous mice to be significantly lower than that of wild-type mice, this study at E14.0 showed no significant difference between the two genotypes. To evaluate the amount of calcification around tooth germs, 3D images of mandibles were reconstructed from the calcein-labeled sections of the wild-type and mutant mice. Labeling was performed at E14.0, and the mice were sacrificed 1 h after the calcein injection to minimize the effect of bone resorption. Comparison of the 3D images revealed that the labeled surface was larger around developing tooth germs in homozygous mouse than in wild-type mouse. On day E15.5, osteoblasts approached the enamel organ of homozygous mice but this was not observed in wild-type mice. In this study, we report a systemic increase in osteoblast number and accelerated bone formation in homozygous PTHrP-knockout mice, both of which contribute to the abnormal tooth development. PMID- 15542036 TI - A scrutiny of matrix metalloproteinases in osteoclasts: evidence for heterogeneity and for the presence of MMPs synthesized by other cells. AB - Genetic diseases and knockout mice stress the importance of matrix metalloproteinases (MMPs) in skeletal turnover. Our study aims at clarifying which MMPs are expressed by osteoclasts. Previous analyses of this basic question led to conflicting reports in the literature. In the present study, we used a variety of approaches: PCR, Northern blots, Slot blots, in situ hybridization, and immunohistochemistry. We analyzed osteoclasts in culture as well as osteoclasts in native bone at different locations and compared mouse and rabbit osteoclasts. Osteoclasts express MMP-9 and -14 in all conditions, although to a variable extent, and they are able to synthesize MMP-3, -10, and -12, at least under some circumstances. The induction of a given MMP in osteoclasts is influenced by its environment (e.g., osteoclast culture vs. native bone, and various sites within the same bone) and depends on the species (e.g., mouse vs. rabbit). Osteoclasts show high amounts of MMP-2 and -13 protein presumably made to a large extent by other cells, thereby documenting how proteinases of nonosteoclastic origin may contribute to osteoclast activities and giving insight in why the resorptive activity of purified osteoclasts appears insensitive to MMP inhibitors. Our study shows that the confusion about osteoclastic MMPs in the literature reflects the remarkable ability of osteoclasts to adapt to their environment, as required by the structural or functional diversity of bone tissue. Our observations provide basic information needed for understanding the emerging role of MMPs in controlling cell signaling and bone resorption. PMID- 15542037 TI - Detection of osteoclastic cell-cell fusion through retroviral vector packaging. AB - Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-kappaB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors. PMID- 15542038 TI - Interactions between estrogen and mechanical strain effects on U2OS human osteosarcoma cells are not influenced by estrogen receptor type. AB - Estrogens (E) and mechanical strain (MS) exert direct effects on osteoblast activity, with good evidence of interactions between their respective effects. Osteoblasts express both forms of estrogen receptors (ER) ERalpha and ERbeta, and previous studies have suggested a specific role for each receptor. Therefore, our working hypothesis was that the interactions between E and MS on osteoblast activity vary depending on which ER is preferentially activated. Using human osteosarcoma cells U2OS stably transfected either with ERalpha or ERbeta, we evaluated the effects of cyclical cell loading on a F-3000 Flexercell Strain Unit (1.5% elongation, 10 min/day) in presence of estradiol (E2) 10(-8) M or not. The original U2OS cell line, which does not express ER, was characterized by low alkaline phosphatase (AP) activity. In both U2OS-ERalpha and U2OS-ERbeta cell lines, MS induced similar increases in AP activity and gene expression as measured by real-time quantitative RT-PCR, and a decrease in type I collagen gene expression. MS and E2 had a synergistic effect on AP activity as compared to each stimulus alone. No change in proliferation rate was observed. Neither proliferation nor differentiation of the original U2OS cell line was altered by strain or E2. In summary, our data showing differences in response to MS between the U2OS with no ER expression and the U2OS-ERalpha or -ERbeta cell lines provide additional evidence that ER plays a critical role in mechanotransduction. However, we were not able to demonstrate that interactions between E and MS were dependent on ER type in U2OS osteosarcoma cells. PMID- 15542039 TI - Effects of vitamin K2 (menatetrenone) and alendronate on bone mineral density and bone strength in rats fed a low-magnesium diet. AB - In this study, we examined changes in bone parameters and bone strength in rats fed low-Mg diets (experiment 1) and the effects of vitamin K2 (MK-4, experiment 3) and alendronate (ALN, experiment 2) in this model. In experiment 1, 5-week-old male Wistar rats were fed three low-Mg diets (Mg 9, 6, 3 mg/100 g diet) for 4 weeks. Although the cortical bone mineral content (CtBMC) and cortical thickness (CtTh) of the femoral diaphysis in all low-Mg-diet groups were the same as or greater than those in the intact group (Mg: 90 mg/100 g diet), the maximum load and elastic modulus were significantly reduced in the 3-mg-Mg group. In experiment 2, 4-week-old Wistar rats were fed a 6-mg-Mg diet for 8 weeks, and the effect of ALN (2, 20, and 200 microg/kg twice a week) was evaluated. The administration of ALN at 200 microg/kg increased the cortical bone mineral content (CtBMC), CtTh, and maximum load, but had no effect on the elastic modulus, as compared with the low-Mg-control group. In experiment 3, the effect of MK-4 was evaluated under the same conditions as in experiment 2. The administration of MK-4 had no effect on CtBMC, CtTh, or bone components of the femoral diaphysis. However, MK-4 inhibited the decreases in maximum load and elastic modulus due to the low-Mg diet. Since there is no other experimental model in which there is a decrease in bone mechanical properties without a decrease in bone mineral content, the low-Mg diet model is considered to be an excellent model for examining bone quality. Our results from this model suggest that MK-4 and ALN affect bone mechanical properties by different mechanisms. PMID- 15542040 TI - Everolimus suppresses cancellous bone loss, bone resorption, and cathepsin K expression by osteoclasts. AB - The proliferation inhibitor of the macrolide class, everolimus, is a drug shown to be effective in the prevention of organ transplant rejection and to have a potential in the treatment of rheumatoid arthritis and certain cancers. As these diseases or their current treatments are associated with bone loss, we examined the effect of everolimus on mouse and human bone cells in vitro and on bone in an ovariectomized (OVX) rat model. Everolimus potently inhibited primary mouse and human osteoclast activity in the pit assay (IC50 values of 0.6-4.0 nM), as well as osteoclast formation, measured as the number of tartrate-resistant acid phosphatase (TRAP) multinucleated cells (IC50 values of 7.7-10.5 nM). Inhibition of osteoblastic differentiation was also observed (IC50 value of 13.5 nM). As expected, everolimus inhibited proliferation of osteoclast precursors and stimulated apoptosis, albeit with insufficient potency and efficacy to explain inhibition of osteoclast activity. Thus, everolimus appeared to directly inhibit bone resorption, which is in accord with the detected inhibition of mRNA and protein expression of cathepsin K; the main collagen-degrading protease in osteoclasts. Despite the in vitro antiproliferative activity of everolimus and the observed inhibition of osteoblast differentiation, no detrimental effects were detected at different skeletal sites in mature OVX rats at doses up to 3 mg/kg/day. This everolimus dose also prevented the OVX-induced loss of cancellous bone by 60%, an effect predominantly associated with decreased osteoclast mediated bone resorption, resulting in a partial preservation of the cancellous bone network. Everolimus inhibited S6 kinase 1 activity in rat blood cells, skin, and bone, at doses equivalent to those used for efficacy experiments in the OVX rat model, which demonstrated in vivo targeting of the expected molecular pathway. In conclusion, everolimus directly inhibits bone resorption by osteoclasts and thus could at least be neutral or protective for bone in vivo, which would favor its use in disease indications associated with bone loss. PMID- 15542041 TI - Heritability of calcaneal quantitative ultrasound measures in healthy adults from the Fels Longitudinal Study. AB - Quantitative ultrasound (QUS) measurements of bone have been reported to predict osteoporotic fracture risk in postmenopausal women and older men. Although many studies have examined the heritability of bone mineral density (BMD), few studies have estimated the heritability of calcaneal QUS phenotypes. In the present study, we examined the genetic regulation of calcaneal QUS parameters in individuals from nuclear and extended families. The study population includes 260 men and 295 women aged 18-91 years (mean+/-SD: 46+/-16 years) who belong to 111 pedigrees in the Fels Longitudinal Study. Three measures of calcaneal structure were collected from both the right and left heel using the Sahara bone sonometer. These measures included broadband ultrasound attenuation (BUA), speed of sound (SOS), and the quantitative ultrasound index (QUI). We used a variance components based maximum likelihood method to estimate the heritability of QUS parameters while simultaneously adjusting for covariate effects. Additionally, we used bivariate extensions of these methods to calculate additive genetic and random environmental correlations among QUS measures. All phenotypes demonstrated statistically significant heritabilities (P<0.0000001). Heritabilities in the right heel (h2+/-SE) were h2=0.59+/-0.10 for BUA, h2=0.73+/-0.09 for SOS, and h2=0.72+/-0.09 for QUI. Similarly, heritabilities for the left heel were h2=0.52+/-0.10, h2=0.75+/-0.10, and h2=0.70+/0.10, respectively. There was evidence for significant genetic and environmental correlations among these six QUS measures. Combinations of QUS measures in the right and left heel demonstrated genetic correlations of 0.94-0.99 and all were significantly different from one indicating at least a partially unique genetic architecture for each of these measures. This study demonstrates that QUS measures of the calcaneus among healthy men and women are heritable, and there are large shared additive genetic effects among all of the traits examined. PMID- 15542042 TI - Treatment with raloxifene for 2 years increases vertebral bone mineral density as measured by volumetric quantitative computed tomography. AB - Volumetric quantitative computed tomography (vQCT), using multiple thin-slice acquisition, measures three-dimensional volumetric bone mineral density (BMD, mg/cm3). vQCT is often used to measure BMD of lumbar vertebrae and may detect early changes in trabecular, cortical, or integral BMD that extend beyond the technical limits of areal dual X-ray absorptiometry (DXA) BMD measurements. The objective of this study was to determine the effect of 2 years of raloxifene (RLX) treatment on several volumetric BMD measures in a subset of postmenopausal women (n=58) enrolled in the Multiple Outcomes of Raloxifene Evaluation (MORE) trial. Patients in this study were randomized to one of three treatment groups: placebo (n=21), RLX 60 mg/day (n=17), or RLX 120 mg/day (n=20), and all patients received daily calcium (500 mg) and vitamin D (400-600 IU) supplementation. Data from the raloxifene treatment groups were pooled for each analysis. Following 2 years of raloxifene treatment, there was a significant percent change from baseline in the vQCT regions of interest (ROIs) of midintegral BMD, total trabecular BMD, and total integral BMD (P<0.05) compared to placebo, while there was no significant change in the spinal DXA BMD measurement. These data provide the first longitudinal assessment by vQCT of changes in vertebral bone density after 2 years of treatment with raloxifene. vQCT appears to be a valuable technique for measuring the effects of raloxifene treatment in this population of postmenopausal women with osteoporosis. PMID- 15542043 TI - Bone mineral density by age, gender, pubertal stages, and socioeconomic status in healthy Lebanese children and adolescents. AB - Gender, ethnicity, and lifestyle factors affect bone mass acquisition during childhood, thus the need for age- and sex-adjusted Z scores using ethnic-specific data for bone mineral density (BMD) measurement. This study aimed at establishing normative data for BMD in healthy Lebanese children and adolescents. Three hundred sixty-three healthy children aged 10 to 17 years (mean+/-SD: 13.1+/-2.0) were studied. BMD, bone mineral content (BMC), and lean mass were measured by dual-energy X-ray absorptiometry (DXA) using a Hologic 4500A device, and apparent volumetric BMD (BMAD) of the lumbar spine and the femoral neck were calculated. BMD, BMC, and BMAD were expressed by age groups and Tanner stages for boys and girls separately. There was a significant effect of age and puberty on all bone parameters, except at the femoral neck BMAD in boys. BMC and BMD were higher at cortical sites in boys, including subtotal body and hip; whereas, in girls, it was higher at a site more enriched in trabecular bone, namely the lumbar spine. At several skeletal sites, girls had significantly higher BMD adjusted for lean mass than boys. By the end of puberty, adolescents had a mean BMD that was 43-66% higher at the lumbar spine and 25-41% higher at cortical sites than pre-pubertal children, depending on the gender. Mean BMD values in the study group were significantly lower (P<0.01) than Western normative values, with Z scores ranging between -0.2 and -1.1. In both genders, children of lower socioeconomic status tended to have lower BMD than those from a higher socioeconomic background. This study allows additional insight into gender dimorphism in mineral accretion during puberty. It also provides a valuable reference database for the assessment of BMD in children with pubertal or growth disorders who are of Middle Eastern origin. PMID- 15542044 TI - Low-level lifetime exposure to cadmium decreases skeletal mineralization and enhances bone loss in aged rats. AB - The effects of low-level lifetime exposure to cadmium (Cd) on the skeleton mineral status and the risk of bone loss in the elderly were studied in an experimental model of human environmental exposure in non-Cd-polluted areas. Young female Wistar rats were exposed to 1 mg Cd/l in drinking water for 24 months. Bone mineral content (BMC), density (BMD) and area of the lumbar spine (L1-L5) and femur, and total skeleton BMD (T-BMD) were measured densitometrically at the baseline and after 6, 12, 18, and 24 months. Prevalence of osteopenia and osteoporosis was evaluated based on the BMD T score and Z score. Osteocalcin (OC) in the serum and total alkaline phosphatase (total ALP) in the serum, cortical and trabecular bone samples as bone formation markers, and C-terminal cross linking telopeptide of type I collagen (CTX) in the serum and urine as bone resorption markers were measured. Calcium (Ca) and Cd concentrations in the serum/blood and urine were determined as well. In the Cd-exposed females, the L1 L5 and femur BMC and BMD at all the studied time points were lower compared to control. The exposure to Cd resulted in lower accumulation of peak bone mass, accelerated osteopenia, and enhanced the prevalence of osteoporosis in aged rats. The effect of Cd was more pronounced at the L1-L5 than at the femur. CTX concentration in the urine was decreased after 6 months and next increased compared to control, whereas the urinary loss of Ca was enhanced during the exposure to Cd. After 24 months of the treatment, the serum total ALP activity and the activity of this enzyme in cortical and trabecular bone decreased and serum CTX concentration increased, whereas the concentrations of OC and Ca were unchanged. The study clearly revealed that low-level lifetime exposure to Cd diminishes the accumulation of bone mass during skeletal growth and influences bone metabolism at maturity causing osteopenia, and enhances the age-related bone loss due to high turnover rate leading in consequence to osteoporosis in aged rats. The results together with our previous findings confirm the hypothesis that environmental exposure to Cd may be a risk factor for skeletal diseases. PMID- 15542045 TI - Bone as a source of FGF23: regulation by phosphate? AB - The identification of FGF23 as a factor involved in several disorders of phosphate regulation and of PHEX as the gene mutated in X-linked Hypophosphatemic Rickets indicates that both these genes may be involved in phosphate homeostasis, although their physiological roles are unclear. In this study, FGF23 mRNA expression was analyzed by real-time RT-PCR and found to be higher in normal human bone than in kidney, liver, thyroid, or parathyroid tissue, while expression in oncogenic osteomalacia tumor tissue was several hundred-fold higher than in bone. Expression of FGF23 mRNA in human osteoblast-like bone cells, quantitated by real-time RT-PCR, increased with increasing extracellular phosphate and was 2-fold higher in cells treated with 2 mM extracellular phosphate compared to 0 mM phosphate treatment. PHEX mRNA expression increased 1.3-fold after treatment with 2 mM phosphate. FGF23 expression in the bone cells increased with increased mineralization over a 20-day treatment period under mineralizing conditions with beta-glycerophosphate, while PHEX expression decreased. The results indicate that FGF23 mRNA expression in bone cells is regulated by extracellular phosphate and by mineralization. These results support proposals that bone may be a source of circulating FGF23 and suggest that FGF23 expression by bone is regulated. PMID- 15542046 TI - TNF-alpha antibodies and osteoprotegerin decrease systemic bone loss associated with inflammation through distinct mechanisms in collagen-induced arthritis. AB - INTRODUCTION: Rheumatoid arthritis (RA) is associated with focal and systemic bone loss involving cytokines such as RANKL and TNF-alpha. RANK-L promotes focal and systemic osteoporosis, whereas osteoprotegerin (OPG) inhibits bone resorption. Although anti-TNF-alpha antibodies (anti-TNF-alpha Ab) decrease joint inflammation and bone erosions, their effects on bone loss are unknown. The aim of this study was to evaluate the effects of OPG and anti-TNF-alpha Ab, separately or in combination, on inflammation and bone remodeling in collagen induced arthritis (CIA), a model of RA. METHODS: DBA/1 mice (n=28) were immunized with bovine type II collagen and treated with OPG-Fc or anti-TNF-alpha Ab or both, or saline. One group of mice (n=7) was not immunized (naive group). Urinary deoxypyridinoline (D-pyr) and whole-body bone mineral density (BMD) were measured at baseline and at sacrifice. Histomorphometric parameters were evaluated at the femoral metaphysis. RESULTS: Anti-TNF-alpha Ab, but not OPG, decreased the clinical arthritis score (P<0.02 vs. saline) and the histological score of inflammation. The BMD change from baseline to sacrifice (DeltaBMD) was significantly smaller in CIA mice than naive mice. OPG and anti-TNF-alpha Ab significantly increased DeltaBMD versus saline, and the effect was greater with OPG (P<0.003). DeltaD-pyr decreased by 65% with OPG and 13% with anti-TNF-alpha Ab. Compared with saline, OPG increased trabecular bone volume (BV/TV) (P<0.02), decreased trabecular separation (P<0.02), and decreased the bone formation rate (BFR) (P<0.01). Anti-TNF-alpha Ab produced no significant changes in bone volume or trabecular separation but increased trabecular thickness (P<0.02 vs. saline) to a value close to that in naive mice, suggesting preservation of bone formation. No additive effects of OPG and anti-TNF-alpha Ab were found. CONCLUSIONS: Systemic OPG and anti-TNF-alpha Ab therapy prevented bone loss in CIA mice through distinct mechanisms involving decreased bone resorption and preserved bone formation. Combining these two agents might help to prevent bone loss in inflammatory diseases. PMID- 15542047 TI - Enhanced bone mass and physical fitness in young female handball players. AB - This study evaluates the effect of physical activity on the bone content (BMC) and density (BMD) in 51 girls (14.2+/-0.4 yr). Twenty-four were placed in the handball group as they have been playing handball for at least 1 year (3.9+/ 0.4). The other 27 who did not perform in any kind of regular physical activity other than that programmed during the compulsory physical education courses comprised the control group. Bone mass and areal density were measured by dual energy X-ray absorptiometry (DXA). The maximal leg extension isometric force in the squat position with knees bent at 90 degrees and the peak force, mean power, and height jumped during vertical squat jump were assessed with a force plate. Additionally, 30-m run (running speed) and 300-m run (as an estimate of anaerobic capacity) tests were also performed. Maximal aerobic capacity was estimated using the 20-m shuttle-run tests. Compared to the controls, handballers attained better results in the physical fitness tests and had a 6% and 11% higher total body and right upper extremity lean mass (all P<0.05). The handballers showed enhanced BMC and BMD in the lumbar spine, pelvic region, and lower extremity (all P<0.05). They also showed greater BMC in the whole body and enhanced BMD in the right upper extremity and femoral neck than the control subjects (all P<0.05). As expected, total lean mass strongly correlated with total and regional BMC and BMD (r=0.79-0.91 P<0.001). Interestingly, 300-m running speed correlated with BMC and BMD variables (r=0.59-0.67 and r=0.60-0.70, respectively; all P<0.001). Multiple regression analysis showed that the 30-m running speed test, combined with the height and body mass, has also predictive value for whole-body BMC and BMD (R=0.93 and R=0.90, P<0.001). In conclusion, handball participation is associated with improved physical fitness, increased lean and bone masses, and enhanced axial and appendicular BMD in young girls. The combination of anthropometric and fitness-related variables may be used to detect girls with potentially reduced bone mass. PMID- 15542048 TI - The effect on cartilage of different forms of application of postmenopausal estrogen therapy: comparison of oral and transdermal therapy. AB - The effect on urine C-telopeptides of type II collagen (uCTX-II) of oral and transdermal estradiol treatment was compared using samples from two randomized, double-blind, placebo-controlled trials. A total of 171 healthy, Danish postmenopausal women, 45-65 years of age completed the 2-year study periods. The uCTX-II marker assessed cartilage degradation, and this response was compared with the effect on urine C-telopeptides of type I collagen (uCTX-I), considered a specific marker of bone resorption. Doses in the oral estradiol treatment groups (continuous combined therapy) were 1 mg 17-beta-estradiol+1 mg drosperinone or 1 mg 17-beta-estradiol+2 mg drosperinone or 1 mg 17-beta-estradiol+3 mg drosperinone or placebo. Doses in the transdermal estradiol treatment groups (continuous combined therapy) were 45 microg 17-beta-estradiol+30 levonorgestrel or 45 microg 17-beta-estradiol+40 microg levonorgestrel or placebo. The effect of oral and transdermal estradiol therapy on cartilage degradation was reflected as a decrease of 19-30% in uCTX-II (P=0.02 and P=0.003 vs. placebo) after 1 year of treatment. uCTX-I decreased 70% (P<0.0001 vs. placebo) reflecting a pronounced effect on bone resorption that was consistent with a 2-year increase in spine and hip BMD of 7-8% and 4-6%, respectively. The results indicate that different regimens of postmenopausal HRT both have an effect on cartilage and bone thus protecting against osteoporosis and osteoarthritis (OA). However, long-term clinical trials are needed to further investigate this issue. PMID- 15542049 TI - Letter to the editor. PMID- 15542051 TI - 2-Phenyl-beta-lapachone can affect mitochondrial function by redox cycling mediated oxidation. AB - 2-Phenyl-beta-lapachone (3,4-dihydro-2-methyl-2-phenyl-2H-naphtho[1,2b]pyran-5,6 dione) (2PBL) is a o-naphthoquinone synthesized as a possible antitumoral agent. The addition of micromolar concentrations of 2PBL to rat liver mitochondria (in the presence of malate-glutamate or succinate, as respiratory substrates): (1) stimulated O(2) consumption in state 4 and inhibited O(2) consumption in state 3, thus decreasing respiratory control index (RCI); and (2) collapsed the mitochondrial membrane potential. The addition of 2PBL to rat liver submitochondrial particles: (1) stimulated NADH oxidation in the presence of rotenone, antimycin, myxothiazol or cyanide; (2) stimulated (.-)O(2)(-) production in the presence of NADH and antimycin; and (3) led to 2PBL semiquinone radical production. Control studies carried out with two p-naphthoquinones, menadione and atovaquone, did not produced equivalent effects. These findings support the hypothesis that 2PBL, undergoes redox cycling and affects mitochondrial function. The 2PBL effect is complex, involving inhibition of electron transfer, uncoupling of oxidative phosphorylation, collapse of mitochondrial membrane potential and (.-)O(2)(-) production by redox cycling. The mitochondrion could be a target organelle for 2PBL cytotoxicity. PMID- 15542052 TI - Enantiospecific (+)- and (-)-germacrene D synthases, cloned from goldenrod, reveal a functionally active variant of the universal isoprenoid-biosynthesis aspartate-rich motif. AB - The naturally occurring, volatile sesquiterpene hydrocarbon germacrene D has strong effects on insect behaviour and genes encoding enzymes that produce this compound are of interest in the study of plant-insect interactions and in a number of biotechnological approaches to pest control. Goldenrod, Solidago canadensis, is unusual in that it produces both enantiomers of germacrene D. Two new sesquiterpene synthase cDNAs, designated Sc11 and Sc19, have been isolated from goldenrod and functional expression in Escherichia coli identified Sc11 as (+)-germacrene D synthase and Sc19 as (-)-germacrene D synthase. Thus, the enantiomers of germacrene D are the products of separate, but closely related (85% amino-acid identity), enzymes. Unlike other sesquiterpene synthases and the related monoterpene synthases and prenyl transferases, which contain the characteristic amino-acid motif DDXX(D,E), Sc11 is unusual in that this motif occurs as (303)NDTYD. Mutagenesis of this motif to (303)DDTYD gave rise to an enzyme that fully retained (+)-germacrene D synthase activity. The converse mutation in Sc19 (D303N) resulted in a less efficient but functional enzyme. Mutagenesis of position 303 to glutamate in both enzymes resulted in loss of activity. These results indicate that the magnesium ion-binding role of the first aspartate in the DDXXD motif may not be as critical as previously thought. Further amino-acid sequence comparisons and molecular modelling of the enzyme structures revealed that very subtle changes to the active site of this family of enzymes are required to alter the reaction pathway to form, in this case, different enantiomers from the same enzyme-bound carbocationic intermediate. PMID- 15542053 TI - A photoaffinity probe designed for host-specific signal flavonoid receptors in phytopathogenic Peronosporomycete zoospores of Aphanomyces cochlioides. AB - Aphanomyces cochlioides zoospores show chemotaxis to cochliophilin A (5-hydroxy 6,7-methylenedioxyflavone, 1), a host derived attractant, and also respond to 5,7 dihydroxyflavone (2) known as an equivalent chemoattractant. To investigate the chemotactic receptors in the zoospores, we designed photoaffinity probes 4'-azido 5,7-dihydroxyflavone (3) and 4'-azido-7-O-biotinyl-5-hydroxyflavone (4) considering chemical structure of 2. Both 3 and 4 had zoospore attractant activity which was competitive with that of 1. When zoospores were treated with the biotinylated photoaffinity probe followed by UV irradiation and streptavidin gold or peroxidase-conjugated streptavidin, probe-labeled proteins were detected on the cell membrane. This result indicated that the 1-specific-binding proteins, a candidate for hypothetical cochliophilin A receptor, were localized on the cell membrane of the zoospores. This is the first experimental evidence of flavonoid binding proteins being present in zoospores, using chemically synthesized azidoflavone as photoaffinity-labeling reagent. PMID- 15542054 TI - Gene expression profiles in rat intestine identify pathways for 1,25 dihydroxyvitamin D(3) stimulated calcium absorption and clarify its immunomodulatory properties. AB - Microarray technology has been used to discover 1,25-dihydroxyvitamin D(3) (1,25 (OH)(2)D(3)) induced gene expression changes in rat small intestine in vivo. Here, we report gene expression changes related to intestinal absorption or transport, the immune system and angiogenesis in response to 1,25-(OH)(2)D(3). Vitamin D deficient rats were intrajugularly given vehicle or vehicle containing 730 ng of 1,25-(OH)(2)D(3)/kg of body weight. Intestinal mRNA was harvested from duodenal mucosa at 15 min, 1, 3, and 6 h post-injection and studied by Affymetrix microarrays. Genes significantly affected by 1,25-(OH)(2)D(3) were confirmed by quantitative RT-PCR with remarkable agreement. The most strongly affected gene in intestine was CYP24 with 97-fold increase at 6 h post-1,25-(OH)(2)D(3) treatment. Intestinal calcium absorption genes: TRPV5, TRPV6, calbindin D(9k), and Ca(2+) dependent ATPase all were up-regulated in response to 1,25-(OH)(2)D(3), supporting the currently accepted mechanism of 1,25-(OH)(2)D(3) induced transcellular calcium transport. However, a 1,25-(OH)(2)D(3) suppression of several intra-/intercellular matrix modeling proteins such as sodium/potassium ATPase, claudin 3, aquaporin 8, cadherin 17, and RhoA suggests a vitamin D regulation of tight junction permeability and paracellular calcium transport. Several other genes related to the immune system and angiogenesis whose expression was changed in response to 1,25-(OH)(2)D(3) provided evidence for an immunomodulatory and anti-angiogenic role of 1,25-(OH)(2)D(3). PMID- 15542055 TI - Altered mitochondrial function and cholesterol synthesis influences protein synthesis in extended HepG2 spheroid cultures. AB - Cultures of hepatocytes and HepG2 cells provide useful in vitro models of liver specific function. In this study, we investigated metabolic and biosynthetic function in 3-D HepG2 spheroid cultures, in particular to characterise changes on prolonged culture. We show that HepG2 cells cultured in spheroids demonstrate a reduction in mitochondrial membrane potential and respiration following 10 days of culture. This coincides with a modest reduction in glycolysis but an increase in glucose uptake where increased glycogen synthesis occurs at the expense of the intracellular ATP pool. Lowered biosynthesis coincides with and is linked to mitochondrial functional decline since low glucose-adapted spheroids, which exhibit extended mitochondrial function, have stable biosynthetic activity during extended culture although biosynthetic function is lower. This indicates that glucose is required for biosynthetic output but sustained mitochondrial function is required for the maintenance of biosynthetic function. Furthermore, we show that cholesterol synthesis is markedly increased in spheroids cf. monolayer culture and that inhibition of cholesterol synthesis by lovastatin extends mitochondrial and biosynthetic function. Therefore, increased cholesterol synthesis and/or its derivatives contributes to mitochondrial functional decline in extended HepG2 spheroid cultures. PMID- 15542056 TI - Mitochondrial and nuclear DNA damage induced by 5-aminolevulinic acid. AB - 5-Aminolevulinic acid (ALA) is a heme precursor accumulated in plasma and in organs in acute intermittent porphyria (AIP), a disease associated with neuromuscular dysfunction and increased incidence of hepatocellular carcinoma (HCC). Liver biopsies of AIP patients showed odd-shaped mitochondria and autophagic vacuoles containing well-preserved mitochondria. ALA yields reactive oxygen species upon metal-catalyzed oxidation and causes in vivo and in vitro impairment of rat liver mitochondria and DNA damage. Using a quantitative polymerase chain reaction assay, we demonstrated that ALA induces a dose dependent damage in nuclear and mitochondrial DNA in human SVNF fibroblasts and rat PC12 cells. CHO cells treated with ALA also show nuclear DNA damage and human HepG2 cells entered in apoptosis and necrosis induced by ALA and its dimerization product, DHPY. The present data provide additional information on the genotoxicity of ALA, reinforcing the hypothesis that it may be involved in the development of HCC in AIP patients. PMID- 15542057 TI - Oxidation of methionine residues in the prion protein by hydrogen peroxide. AB - Reaction of H(2)O(2) with the recombinant SHa(29-231) prion protein resulted in rapid oxidation of multiple methionine residues. Susceptibility to oxidation of individual residues, assessed by mass spectrometry after digestion with CNBr and lysC, was in general a function of solvent exposure. Met 109 and Met 112, situated in the highly flexible amino terminus, and key residues of the toxic peptide PrP (106-126), showed the greatest susceptibility. Met 129, a residue located in a polymorphic position in human PrP and modulating risk of prion disease, was also easily oxidized, as was Met 134. The structural effect of H(2)O(2)-induced methionine oxidation on PrP was studied by CD spectroscopy. As opposed to copper catalyzed oxidation, which results in extensive aggregation of PrP, this reaction led only to a modest increase in beta-sheet structure. The high number of solvent exposed methionine residues in PrP suggests their possible role as protective endogenous antioxidants. PMID- 15542058 TI - The lipocalin alpha(1)-microglobulin binds heme in different species. AB - The lipocalin alpha(1)-microglobulin (alpha(1)m), found in plasma and tissues of various vertebrates, is brown, forms complexes with other proteins and has immunomodulatory effects in vitro, but the physiological function is not yet established. Human alpha(1)m was recently shown to bind heme and, after cleavage of a C-terminal tetrapeptide, initiate heme degradation, thus suggesting a heme scavenger function. In this work the heme-binding of alpha(1)m was characterized using heme immobilized on agarose beads, spectrophotometry, and electrophoresis. alpha(1)m, both in plasma and in purified form, displayed a concentration dependent binding to heme-agarose. The apparent dissociation-constant was estimated to be around 2 x 10(-6)M for both free alpha(1)m and the IgA-alpha(1)m complex. Incubation with free heme resulted in two forms of alpha(1)m with different electrophoretic mobility. alpha(1)m, identified on Western blotting, was found in eluates from heme-agarose after incubation with human biological fluids as well as sera from non-human species, indicating evolutionary conservation of the heme-binding property. Heme-binding could be instrumental for isolating new alpha(1)m-homologues. PMID- 15542059 TI - The cattle tick antigen, Bm95, expressed in Pichia pastoris contains short chains of N- and O-glycans. AB - Bm95 is an antigen isolated from Boophilus microplus strains with low susceptibility to antibodies developed in cattle vaccinated with the recombinant Bm86 antigen (Gavac, HeberBiotec S.A., Cuba). It is a Bm86-like surface protein, which by similarity contains seven EGF-like domains and a lipid-binding GPI anchor site at the C-terminal region. The primary structure of the recombinant (rBm95) protein expressed in Pichia pastoris was completely verified by LC/MS. The four potential glycosylation sites (Asn 122, 163, 329, and 363) are glycosylated partially with short N-glycans, from Man(5)GlcNAc(2) to Man(9)GlcNAc(2) of which, Man(8-9)GlcNAc(2) were the most abundant. O Glycopeptides are distributed mostly towards the protein N-terminus. While the first N-glycosylated site (Asn(122)) is located between EGF-like domains 2 and 3, where the O-glycopeptides were found, two other N-glycosylated sites (Asn(329) and Asn(363)) are located between EGF-like domains 5 and 6, a region devoid of O glycosylated Ser or Thr. PMID- 15542060 TI - Purification, physicochemical characterization, saccharide specificity, and chemical modification of a Gal/GalNAc specific lectin from the seeds of Trichosanthes dioica. AB - A new galactose-specific lectin has been purified from the extracts of Trichosanthes dioica seeds by affinity chromatography on cross-linked guar gum. The purified lectin (T. dioica seed lectin, TDSL) moved as a single symmetrical peak on gel filtration on Superose-12 in the presence of 0.1 M lactose with an M(r) of 55 kDa. In the absence of ligand, the movement was retarded, indicating a possible interaction of the lectin with the column matrix. In SDS-PAGE, in the presence of beta-mercaptoethanol, two non-identical bands of M(r) 24 and 37 kDa were observed, whereas in the absence of beta-mercaptoethanol, the lectin yielded a single band corresponding to approximately 55,000 Da, indicating that the two subunits of TDSL are connected by one or more disulfide bridges. TDSL is a glycoprotein with about 4.9% covalently bound neutral sugar. Analysis of near-UV CD spectrum by three different methods (CDSSTR, CONTINLL, and SELCON3) shows that TDSL contains 13.3% alpha-helix, 36.7% beta-sheet, 19.4% beta-turns, and 31.6% unordered structure. Among a battery of sugars investigated, TDSL was inhibited strongly by beta-d-galactopyranosides, with 4-methylumbelliferyl-beta-d galactopyranoside being the best ligand. Chemical modification studies indicate that tyrosine residues are important for the carbohydrate-binding and hemagglutinating activities of the lectin. A partial protection was observed when the tyrosine modification was performed in the presence of 0.2 M lactose. The tryptophan residues of TDSL appear to be buried in the protein interior as they could not be modified under native conditions, whereas upon denaturation with 8 M urea two Trp residues could be selectively modified by N-bromosuccinimide. The subunit composition and size, secondary structure, and sugar specificity of this lectin are similar to those of type-2 ribosome inactivating proteins, suggesting that TDSL may belong to this protein family. PMID- 15542061 TI - Macrophage-derived peroxynitrite diffusion and toxicity to Trypanosoma cruzi. AB - We studied the capacity of macrophage-derived peroxynitrite to diffuse into and exert cytotoxicity against Trypanosoma cruzi, the causative agent of Chagas' disease. In two types of macrophage-T. cruzi co-cultures, one with a fixed separation distance between source and target cells, and another involving cell to-cell interactions, peroxynitrite resulted in significant oxidation of intracellular dihydrorhodamine and inhibition of [(3)H]thymidine incorporation in T. cruzi, which were not observed by superoxide or nitric oxide alone. The effects were attenuated in the presence of bicarbonate, in agreement with the extracellular consumption of peroxynitrite by its fast reaction with CO(2). However, studies using different T. cruzi densities, which allow to modify average diffusion distances of exogenously added peroxynitrite to target cells, indicate that at distances <5 microm, the diffusion process outcompetes the reaction with CO(2) and that the levels of peroxynitrite formed by macrophages would be sufficient to cause toxicity to T. cruzi during cell-to-cell contact and/or inside the phagosome. PMID- 15542062 TI - The reaction of indole with the aminoacrylate intermediate of Salmonella typhimurium tryptophan synthase: observation of a primary kinetic isotope effect with 3-[(2)H]indole. AB - The bacterial tryptophan synthase alpha(2)beta(2) complex catalyzes the final reactions in the biosynthesis of L-tryptophan. Indole is produced at the active site of the alpha-subunit and is transferred through a 25-30 A tunnel to the beta active site, where it reacts with an aminoacrylate intermediate. Lane and Kirschner proposed a two-step nucleophilic addition-tautomerization mechanism for the reaction of indole with the aminoacrylate intermediate, based on the absence of an observed kinetic isotope effect (KIE) when 3-[(2)H]indole reacts with the aminoacrylate intermediate. We have now observed a KIE of 1.4-2.0 in the reaction of 3-[(2)H]indole with the aminoacrylate intermediate in the presence of monovalent cations, but not when an alpha-subunit ligand, disodium alpha glycerophosphate (Na(2)GP), is present. Rapid-scanning stopped flow kinetic studies were performed of the reaction of indole and 3-[(2)H]indole with tryptophan synthase preincubated with L-serine, following the decay of the aminoacrylate intermediate at 350 nm, the formation of the quinonoid intermediate at 476 nm, and the formation of the L-Trp external aldimine at 423 nm. The addition of Na(2)GP dramatically slows the rate of reaction of indole with the alpha-aminoacrylate intermediate. A primary KIE is not observed in the reaction of 3-[(2)H]indole with the aminoacrylate complex of tryptophan synthase in the presence of Na(2)GP, suggesting binding of indole with tryptophan synthase is rate limiting under these conditions. The reaction of 2-methylindole does not show a KIE, either in the presence of Na(+) or Na(2)GP. These results support the previously proposed mechanism for the beta-reaction of tryptophan synthase, but suggest that the rate limiting step in quinonoid intermediate formation from indole and the aminoacrylate intermediate is deprotonation. PMID- 15542063 TI - The iSH2 domain of PI 3-kinase is a rigid tether for p110 and not a conformational switch. AB - Class IA PI 3-kinases are heterodimeric proteins with distinct catalytic (p110) and regulatory (p85) subunits. The minimal fragment of p85 capable of regulating p110 activity (p85ni) is the N-terminal SH2 domain linked to the iSH2 coiled-coil domain. We used cysteine mutagenesis and (14)C-NEM-labeling to show that the p110 binding site in the iSH2 domain includes two regions: residues 482-484 and 532 541. These regions are adjacent to each other in the three-dimensional structural model of the iSH2 domain, and define a coherent binding site. We then used spin labeling and EPR spectroscopy to demonstrate that the conformation of the iSH2 domain is unaffected by binding to the N-terminal fragment of p110 (residues 1 108), and/or by phosphopeptide binding to p85ni/p110(1-108) heterodimers. Finally, we show that the cSH2 domain cannot substitute for the nSH2 domain with regard to inhibition of p110. These data support a model in which the iSH2 domain is a rigid tether for p110, and regulation of p85/p110 is mediated by nSH2-p110 contacts. PMID- 15542064 TI - Diallyl sulfide induces heme oxygenase-1 through MAPK pathway. AB - Diallyl sulfide (DAS), is protective against chemically induced heptotoxicity, mutagenesis, and carcinogenesis. The mechanism of its protective effects is not fully understood. In this study, we found that DAS can induce the expression of heme oxygenase-1 (HO-1), which plays a critical role in the cell defense system against oxidative stress. DAS causes a dose- and time-dependent increase of HO-1 protein and mRNA level without toxicity in HepG2 cells. DAS-induced HO-1 protein expression is dependent on newly synthesized mRNA and newly synthesized protein. DAS increases Nrf2 protein expression, nuclear translocation, and DNA-binding activity. The MAP kinase ERK is activated by DAS. Both ERK and p38 pathways play an important role in DAS-induced Nrf2 nuclear translocation and ho-1 gene activation. DAS stimulates a transient increase of reactive oxygen species (ROS). N-Acetyl-cysteine blocked this increase of ROS production as well as DAS-induced ERK activation, Nrf2 protein expression and nuclear translocation, and ho-1 gene activation. The increase in HO-1 produced by DAS protected the HepG2 cells against toxicity by hydrogen peroxide or arachidonic acid. These results suggest that DAS induces ho-1 through production of ROS, and Nrf2 and MAPK (ERK and p38) mediate this induction. Induction of ho-1 may play a role in the protective effects of DAS. PMID- 15542065 TI - Store-operated Ca(2+) entry and tyrosine kinase pp60(src) hyperactivity are modulated by hyperglycemia in platelets from patients with non insulin-dependent diabetes mellitus. AB - We have investigated the involvement of store-operated Ca(2+) entry (SOCE) in the abnormal platelet Ca(2+) homeostasis in patients with non insulin-dependent diabetes mellitus (NIDDM). In a medium containing 180 mg/dL glucose, platelets from NIDDM patients showed an increased SOCE compared to controls. We found that tyrosine phosphorylation was elevated in platelets from NIDDM patients. Consistent with this, the activity of the tyrosine kinase pp60(src) is enhanced in platelets from diabetic patients. When the experiments were performed in a medium containing 90 mg/dL both, SOCE and pp60(src) activity, were similar to those found in control platelets. Our results indicate that SOCE is altered in platelets from NIDDM patients probably due to the increased activity of the tyrosine kinase pp60(src). Both, SOCE and pp60(src) activity in platelets from NIDDM patients are more susceptible to the extracellular glucose concentration, which seems to be involved in the dysfunction of these mechanisms. PMID- 15542066 TI - Twitchin, a thick-filament protein from molluscan catch muscle, interacts with F actin in a phosphorylation-dependent way. AB - Twitchin belongs to the titin-like giant proteins family, it is co-localized with thick filaments in molluscan catch muscles and regulates the catch state depending on its level of phosphorylation. The mechanism by which twitchin controls the catch state remains to be established. We report for the first time the ability of twitchin to interact with F-actin. The interaction is observed at low and physiological ionic strengths, irrespective of the presence or absence of Ca(2+). It was demonstrated by viscosity and turbidity measurements, low- and high-speed co-sedimentation, and with the light-scattering particle size analysis revealing the specific twitchin-actin particles. The twitchin-actin interaction is regulated by twitchin phosphorylation: in vitro phosphorylated twitchin does not interact with F-actin. We speculate that the catch muscle twitchin might provide a mechanical link between thin and thick filaments, which contributes to catch force maintenance. PMID- 15542067 TI - Fluoxetine-treated male wrasses exhibit low AVT expression. AB - In many species, increasing serotonergic activity can reduce aggression and reverse dominance relationships. These effects may in part be mediated through interactions with the arginine vasotocin/vasopressin (AVT/AVP) system. We tested this hypothesis in a territorial coral reef fish, the bluehead wrasse (Thalassoma bifasciatum), by experimentally enhancing serotonergic neurotransmission, using the selective serotonin re-uptake inhibitor (SSRI) fluoxetine. Terminal phase (TP) males received 2 weeks of nightly intraperitoneal fluoxetine injections (6 microg/g body weight) and were then tested for their aggressive response to an intruder and killed to examine AVT phenotype in the preoptic area of the hypothalamus (POA), an area important to social behavior in fishes. Our previously published study demonstrated that fluoxetine-treated males are less aggressive [H.A.N. Perreault, K. Semsar, J. Godwin, Fluoxetine treatment decreases territorial aggression in a coral reef fish, Physiol. and Behav. 79 (2003) 719-724.]. Here, further study of these same fluoxetine-treated males shows approximately twofold lower AVT mRNA expression relative to saline-treated controls in all regions of the POA (all p< or =0.05) without any changes in AVT ir soma size (all p>0.4). This study experimentally supports the hypothesis that behavioral effects of SSRIs may be mediated in part through interactions with the AVT/AVP system. These results parallel findings from rodents and humans and are consistent with an indirect neurosteroidogenic rather than a solely direct serotonergic mechanism for SSRI effects on the AVT/AVP system. Furthermore, they suggest that SSRI effects on neuroendocrine function may be best modeled in animals with sensitive stress responses such as those found in nondomesticated animals. PMID- 15542068 TI - Nitric oxide mediates an LPS-induced depression of cytochrome P450 (CYP1A) activity in astrocytes. AB - During inflammatory responses in the brain, the expression of cytochrome P450 isoforms in the CNS are modulated and the capacity of the brain to metabolize drugs and to synthesize or degrade certain endogenous chemicals and drugs is diminished. While this response can be attributed in part, to the production and action of cytokines within the brain, it is also likely that other inflammatory mediators play an integral role. This paper investigates a potential role for nitric oxide (NO) in the loss of cytochrome P450 (CYP1A) in the brain during inflammation. Escherichia coli lipopolysaccharide (LPS), a commonly used proinflammatory endotoxin, was incubated with cultured rat astrocytes to provide a model of inflammation in the CNS. CYP1A activity was significantly decreased in cultured astrocytes incubated with LPS for 24 h. This loss in enzyme activity was accompanied by a substantial production of nitric oxide (NO) by these cells. Immunohistochemical examination demonstrated an upregulation of inducible nitric oxide synthase (iNOS) expression following the exposure of astrocytes to LPS. The addition of a selective iNOS blocker (1400W) caused a partial but significant reversal of the LPS-mediated loss in CYP1A. The incubation of astrocytes with the NO-generating compound (DETA NONOate) resulted in a loss of CYP1A. Taken together, these observations suggest that NO plays a pivotal role in the inflammation mediated loss in CYP1A activity in the brain. PMID- 15542069 TI - Signal transmission from cones to amacrine cells in dark- and light-adapted tiger salamander retina. AB - Amacrine cells (ACs) are third-order interneurons in the retina that mediate antagonistic surround inputs to retinal ganglion cells and motion-related signals in the inner retina. Previous studies have revealed that rod-to-AC signals in dark-adapted retina are mediated by a nonlinear high-gain synaptic pathway. In this study, we investigated how cone signals are transmitted to ACs under dark- and light-adapted conditions. By using the spectral subtraction method, we found that the voltage gain of the cone-AC synaptic pathway in dark-adapted salamander retina (GD) is between 28 and 72, which is about one order of magnitude lower than the voltage gain of the rod-AC pathway. This suggests that, in darkness, rod signals are more efficiently transmitted to the ACs than cone signals. The voltage gain of the cone-AC synaptic pathway in the presence of 500 nm/-2.4 background light, GL, ranges between 28 and 56. Linear regression analysis indicates that GD and GL are strongly, positively, and linearly correlated. The average GL/GD ratio is 0.73, suggesting that, on average, GL in any given AC is about 73% of GD. This adaptation-induced change in cone-AC voltage gain exemplifies use-dependent modulations of synaptic transmission in the retina, and possible mechanisms underlying light-mediated alterations of retinal synaptic function are discussed. PMID- 15542070 TI - Myelinated nerve fibres in the subcortical white matter of cerebral hemispheres are preserved in alcoholic subjects. AB - Previous studies have found that the primary change in the brain of chronic alcoholics is a selective loss of white matter in cerebral hemispheres. It is unknown whether the loss of white matter is due to alcohol-induced degeneration of axons in white matter, and this hypothesis has never been tested. We used a newly developed stereological method to investigate whether the length and diameter of myelinated fibres change in the subcortical white matter of male alcoholic subjects. We found no significant differences between control and alcoholic subjects with respect to the total volume, total length, and mean diameter of myelinated fibres in white matter. Our results do not find any support for the hypothesis that chronic alcoholic subjects may suffer from a degeneration of axons (myelinated fibres) in white matter in cerebral hemispheres. PMID- 15542071 TI - Elevated voltage-gated Ca2+ channel immunoreactivities in the hippocampus of seizure-prone gerbil. AB - In present study, we investigated voltage-gated Ca2+ channel (VGCC) expressions in the hippocampus of the Mongolian gerbil and its association with different sequelae of spontaneous seizures, in an effort to identify the epileptogenesis in this animal. In the hippocampus of pre-seizure seizure sensitive (SS) gerbils, VGCC subunit expressions were significantly elevated, as compared with seizure resistant (SR) gerbils. In 3 h postictal group, the alteration of VGCC expressions showed regional- and neuronal-specific manners; VGCC immunoreactivities in principal neurons were markedly decreased; however, their immunoreactivities in interneurons were significantly elevated. These results are the first comprehensive description of the distribution of VGCC immunoreactivities in the normal and epileptic hippocampus of gerbils, and suggest that these alterations in the hippocampus of the SS gerbil may be related with tissue excitability and have a role in modulating recurrent excitation following seizures. PMID- 15542072 TI - Neuroprotective properties of catalpol in transient global cerebral ischemia in gerbils: dose-response, therapeutic time-window and long-term efficacy. AB - The present study evaluated for the first time the dose-effectiveness, therapeutic time-window and long-term efficacy of the neuroprotection of catalpol by behavioral and histological measures in gerbils subjected to transient global cerebral ischemia. Catalpol (1 mg/kg ip) used immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h significantly rescued neurons in the hippocampal CA1 subfield and reduced cognitive impairment. The neuroprotective efficacy of catalpol became more evident at the doses of 5 and 10 mg/kg. Of great importance were the findings that the neuroprotective efficacy of catalpol still could be seen even when the treatment was delayed 3 h and when the observational period was lasted out 35 days after ischemia. It was reasonable to draw the conclusion that catalpol was truly neuroprotective rather than simply delayed the onset of neuronal damage. These results suggested that catalpol might be of therapeutic value for global cerebral ischemia. PMID- 15542073 TI - Tamoxifen increases methamphetamine-evoked dopamine output from superfused striatal tissue fragments of male mice. AB - The antiestrogen, tamoxifen (TMX), has been shown to function as a neuroprotectant against the nigrostriatal dopaminergic (NSDA) neurotoxin, methamphetamine (MA), within male mice. In the present report, we examined the effects of a combined infusion of TMX and MA within superfused striatal tissue fragments of male mice as an approach to understand some of the bases for TMX to function as a NSDA neuroprotectant within male mice. In Experiment 1, a coinfusion of TMX at 1, 10, or 100 pg/ml were all equally effective in increasing MA-evoked dopamine (DA) output as compared with a 0 pg/ml (control) dose. In Experiment 2, we tested whether this effect of TMX was specific for MA-evoked DA output by coinfusing TMX with a depolarizing concentration of potassium chloride (K+ -30 mM). No statistically significant differences were obtained between superfusions of striatal tissue fragments stimulated with K+ in the presence or absence of TMX (100 pg/ml). In Experiment 3, we assessed whether these effects of TMX may be exerted upon the dopamine transporter (DAT) by coinfusing DA (1 microM) in the presence or absence of TMX (100 pg/ml). No differences in DA recovery rates were obtained between superfusions performed in the presence or absence of TMX. Taken together, these results show that the striatum of male mice is very sensitive to the modulatory effects of TMX upon MA-evoked DA output. These effects of TMX appear to be relatively specific for MA-evoked DA output, as K+ -stimulated DA was not altered by TMX, and do not appear to exert these effects by altering dopamine transporter function. PMID- 15542074 TI - Effect of intermittent hypoxia on long-term potentiation in rat hippocampal slices. AB - Intermittent hypoxia (IH) during sleep has been shown to induce apoptosis in a time-dependent manner and spatial learning deficits in adult rats. Recently, we have demonstrated that IH induced significant decreases in Ser-133-phosphorylated cAMP-response element-binding protein (pCREB) without changes in total CREB. The expression of cleaved caspase 3 in the hippocampal CA1, a marker of apoptosis, peaked at 3 days of IH and returned to normoxic values at 14 days of IH. In addition, biphasic changes in spatial task learning were correlated with the CREB phosphorylation time course. In the present study, the rat hippocampal slice preparation was used to evaluate the ability to induce and maintain a CA1 population spike long-term potentiation (PS-LTP) in room air (RA)-maintained and IH-exposed rats. A significant decrease in the ability to sustain PS-LTP for 15 min in slices prepared from IH-exposed rats for either 3 days (34% of total) or 7 days (51% of total) as compared to slices prepared from RA-maintained rats (76% of total) was observed. These results suggest that the diminishment in the ability of neuronal tissue to express and sustain PS-LTP is correlated with previously reported biphasic changes in CREB phosphorylation and programmed cell death. PMID- 15542075 TI - Neuroprotective effects of the free radical scavenger Edaravone (MCI-186) in mice permanent focal brain ischemia. AB - The present study was aimed to evaluate the effect of the free radical scavenger Edaravone on infarct volume due to permanent MCA occlusion in mice and, if so, to elucidate the mechanism of its neuroprotective effects. Male Balb/c mice were subjected to permanent middle cerebral artery occlusion and were treated with 3.0 mg/kg of Edaravone or vehicle 30 min before ischemia. Infarct volume was assessed by 2,3,5-triphenyltetrazolium chloride (TTC) method after 24 h. Furthermore, in situ detection of superoxide in the ipsilateral neocortex was carried out using the superoxide-sensitive dye dihydroethidium (DHE) staining technique. Pretreatment with 3.0 mg/kg of Edaravone ameliorated the tissue damage in the infarct rim and significantly reduced infarct volume to about 77% of the control (p<0.05). Semi-quantitative measurement of red fluorescence emitted from DHE revealed that the superoxide increased in the ischemic core at 1 h after the onset of ischemia and extended towards the infarct rim at 3 and 6 h, and that pretreatment with 3.0 mg/kg of Edaravone significantly inhibited the increase of superoxide in the infarct rim at 3 and 6 h (p<0.01). Double staining with DHE and monoclonal antibody against NeuN showed that the majority of the nuclei positive for DHE were also positive for NeuN. These findings suggest that Edaravone salvages the boundary zone of infarct by scavenging reactive oxygen species especially in the neurons during permanent focal cerebral ischemia. PMID- 15542076 TI - Distribution of alpha1, alpha4, gamma2, and delta subunits of GABAA receptors in hippocampal granule cells. AB - GABAA receptors are pentamers composed of subunits derived from the alpha, beta, gamma, delta, theta, epsilon, and pi gene families. alpha1, alpha4, gamma2, and delta subunits are expressed in the dentate gyrus of the hippocampus, but their subcellular distribution has not been described. Hippocampal sections were double labeled for the alpha1, alpha4, gamma2, and delta subunits and GAD65 or gephyrin, and their subcellular distribution on dentate granule cells was studied by means of confocal laser scanning microscopy (CLSM). The synaptic versus extrasynaptic localization of these subunits was inferred by quantitative analysis of the frequency of colocalization of various subunits with synaptic markers in high resolution images. GAD65 immunoreactive clusters colocalized with 26.24+/-0.86% of the alpha1 subunit immunoreactive clusters and 32.35+/-1.49% of the gamma2 subunit clusters. In contrast, only 1.58+/-0.13% of the alpha4 subunit immunoreactive clusters and 1.92+/-0.15% of the delta subunit clusters colocalized with the presynaptic marker GAD65. These findings were confirmed by studying colocalization with immunoreactivity of a postsynaptic marker, gephyrin, which colocalized with 27.61+/-0.16% of the alpha1 subunit immunoreactive clusters and 23.45+/-0.32% of the gamma2 subunit immunoreactive clusters. In contrast, only 1.90+/-0.13% of the alpha4 subunit immunoreactive clusters and 1.76+/-0.10% of the delta subunit clusters colocalized with gephyrin. These studies demonstrate that a subset of alpha1 and gamma2 subunit clusters colocalize with synaptic markers in hippocampal dentate granule cells. Furthermore, all four subunits, alpha1, alpha4, gamma2, and delta, are present in the extrasynaptic locations. PMID- 15542077 TI - Evaluation of the tail formalin test in mice as a new model to assess local analgesic effects. AB - Opioids are effective topical analgesics in the radiant heat tailflick assay and display synergistic interactions with a number of other classes of drugs. To determine whether these actions extend to other types of nociception, we examined the actions of topical morphine and lidocaine in a tail formalin assay in the mouse. Formalin responses in the tail were similar to those seen in the hind paw, but were limited to licking. Unlike the traditional hind paw assay, the time course of nociceptive behavior in the tail was monophasic; lasting 40-60 min. Morphine, MK-801 and acetylsalicylic acid (ASA) were active systemically in the tail formalin assay with potencies similar to those seen in the second phase of the paw formalin test. Both morphine and lidocaine were active topically in the tail formalin assay, although their time-course of action appeared to be shorter than that of the formalin. However, morphine displayed ceiling effect not seen when it was administered systemically. Lidocaine also had a ceiling effect. When given together, the response to the combination was supra-additive, consistent with our prior studies showing synergy in the radiant heat tailflick assay. These studies validate the formalin assay in the tail and support the topical actions of opioids and other drugs in a second pain model. They also suggest supra additive interactions between morphine and lidocaine similar to those previously seen. The tail formalin assay will be valuable in assessing the activity of topical drugs. PMID- 15542078 TI - Electrophysiological and neurochemical characterization of neurons of the medial preoptic area in Japanese quail (Coturnix japonica). AB - Intracellular recordings of medial preoptic neurons demonstrated that most neurons show a spontaneous firing, a linear I-V relationship and low-threshold like events suppressed by the application of Ni2+. Some neurons had a depolarizing sag of the membrane potential in response to hyperpolarizing current pulses. The majority of the cells exhibited a robust spontaneous synaptic activity suppressed by SR95531 (100 microM), a GABAA receptor antagonist, and/or by 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), an (RS)-alpha-amino-3 hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/kainate (KA) glutamate receptor antagonist. Most neurons were affected by the application of AMPA (10 microM), kainate (30 microM), N-methyl-D-aspartic acid (NMDA, 10 microM), isoguvacine (a GABAA receptor agonist, 100 microM), dopamine (100 microM), and norepinephrine (100 microM). Biocytin injections coupled to aromatase immunocytochemistry identified 19 recorded neurons including 3 displaying a dense aromatase immunoreactivity. All of them responded to kainate, dopamine, and norepinephrine, while only one responded to isoguvacine and NMDA. Taken together, these results demonstrate a relative electrical and neurochemical homogeneity of the medial preoptic neurons, including a few aromatase-immunoreactive neurons that could be identified by immunocytochemistry after biocytin labeling of the recorded neurons. PMID- 15542079 TI - Binding characteristics and daily rhythms of melatonin receptors are distinct in the retina and the brain areas of the European sea bass retina (Dicentrarchus labrax). AB - Melatonin is synthesized, with a circadian rhythm, in the pineal organ of vertebrates, high levels being produced during the scotophase and low levels during the photophase. The retina also produces melatonin, although in the case of the European sea bass, its secretion pattern appears to be inverted. In the study described here, radioreceptor assay techniques were used to characterize the melatonin binding sites, their regional distribution and their daily variations. Brain and retina membrane preparations were used in all the binding assays and 2-[125I]iodomelatonin ([125I]Mel) as radioligand at 25 degrees C. The specific binding of [125I]Mel was seen to be saturable, reversible, specific and of high affinity. In all the tissues assayed, the power of the ligands to inhibit [125I]Mel binding decreased in the following order: melatonin>>4-P PDOT>luzindole> or =N-acetylserotonin, which points to the presence of Mel1-like receptors. The inhibition curves of 4-P-PDOT suggested the presence of two different binding sites in the brain areas, but only one type of site of low affinity in the neural retina. No daily variations in [125I]Mel binding capacity (Bmax) or affinity (Kd) were detected in the brain areas, while a clear rhythm in Kd melatonin receptor affinity and Bmax binding capacity was observed in the retina. Kd and Bmax retinal rhythms were out of phase with the lowest Kd and the highest Bmax occurring at scotophase. This result suggests that retinal melatonin is a paracrine factor able to control receptor desensitization during photophase when ocular melatonin is higher in this species. PMID- 15542080 TI - Tetrodotoxin-sensitive and -resistant Na+ channel currents in subsets of small sensory neurons of rats. AB - Voltage-activated Na+ channels in the primary sensory neurons are important for generation of action potentials and regulation of neurotransmitter release. The Na+ channels expressed in different types of dorsal root ganglion (DRG) neurons are not fully known. In this study, we determined the possible difference in tetrodotoxin-sensitive (TTX-S) and -resistant (TTX-R) Na+ channel currents between isolectin B4 (IB4)-positive and IB4-negative small DRG neurons. Whole cell voltage- and current-clamp recordings were performed in acutely isolated DRG neurons labeled with and without IB4 conjugated to Alexa Fluor 594. The peak Na+ current density was significantly higher in IB4-negative than IB4-positive DRG neurons. While all the IB4-negative neurons had a prominent TTX-S Na+ current, the TTX-R Na+ current was present in most IB4-positive cells. Additionally, the evoked action potential had a higher activation threshold and a longer duration in IB4-positive than IB4-negative neurons. TTX had no effect on the evoked action potential in IB4-positive neurons, but it inhibited the action potential generation in about 50% IB4-negative neurons. This study provides complementary new information that there is a distinct difference in the expression level of TTX-S and TTX-R Na+ channels between IB4-negative than IB4-positive small diameter DRG neurons. This difference in the density of TTX-R Na+ channels is responsible for the distinct membrane properties of these two types of nociceptive neurons. PMID- 15542081 TI - No facilitation of amphetamine- or cocaine-induced hyperactivity in adult rats after various 192 IgG-saporin lesions in the basal forebrain. AB - Lesions of basal forebrain cholinergic neurons by intracerebroventricular (i.c.v.) injections of 192 IgG-saporin increased the locomotor response to 0.5 and 1.5 mg/kg of D-amphetamine in adult rats [A. Mattsson, S.O. Ogren, L. Olson, Facilitation of dopamine_mediated locomotor activity in adult rats following cholinergic denervation, Exp Neurol. 174 (2002) 96-108.]. In the present study, adult male rats were subjected to bilateral injections of 192 IgG-saporin either into the septum (Sp), the nucleus basalis magnocellularis (Nbm), both structures (SpNbm) or i.c.v. Locomotor activity was assessed in the home cage 23 days after surgery, and, subsequently, thrice after an intraperitoneal injection of D amphetamine (1 mg/kg) and twice after an injection of cocaine (15 mg/kg). Analysis of AChE-stained material showed that Sp lesions induced preferentially hippocampal denervation, Nbm lesions induced preferentially cortical denervation, while both SpNbm and i.c.v. lesions deprived the hippocampus and the cortex of almost all AChE-positive reaction products. The spontaneous and drug-induced locomotor activity of all lesioned rats did not differ significantly from that of control rats, except in rats subjected to i.c.v. injections, in which the locomotor response was significantly increased after the second administration of cocaine. In addition, in Nbm and SpNbm rats, the locomotor reaction to cocaine was weaker right after the second injection. The present results do not confirm the report by Mattsson et al. on the potentiation of amphetamine-induced locomotion by i.c.v. injections of 192 IgG-saporin, but suggest that cocaine induced locomotion can be increased by such lesions and, to some respect, attenuated by cholinergic damage in the Nbm. PMID- 15542082 TI - Load- and cadence-dependent modulation of somatosensory evoked potentials and Soleus H-reflexes during active leg pedaling in humans. AB - Modulation of transmission in group I muscle afferent pathways to the somatosensory cortex and those to the alpha-motoneuron were investigated during active leg pedaling. Cerebral somatosensory evoked potentials (SEPs) and Soleus (Sol) H-reflexes following posterior tibial nerve stimulation were recorded at four different pedaling phases. The subjects were asked to perform pedaling at three different cadences (30, 45 and 60 rpm with 0.5 kp, cadence task; C-task) and with three different workloads (at 45 rpm with 0.0, 0.5 and 1.0 kp, load task; L-task). In both C- and L-tasks, Sol H-reflexes were modulated in a phase dependent manner, showing an increase in the power phase and a decrease in the recovery phase. In contrast, the early SEP (P30-N40) components were modulated in a phase-dependent manner when the cadence and load were low. When focusing on the power phases, significant cadence- and load-dependent modulations of the P30-N40 were found, and inversely graded with the cadence and load. The H-reflex was found to be significantly decreased at the highest cadence, i.e., cadence dependent modulation. In contrast, the H-reflex during the L-task was found to be proportional to the load. The correlation analysis between the size of H-reflex and the amount of background (BG) electromyographic (EMG) activity demonstrated that the H-reflex in the power phase did not depend on the BG EMG in either C- or L-task. These findings suggested that transmission of muscle afferents along the ascending pathways to the cerebral cortex and the spinal cord is independently controlled in accordance with the biomechanical constraints of active pedaling. PMID- 15542083 TI - Indium-promoted Barbier-type allylations in aqueous media: a convenient approach to 4-C-branched monosaccharides and (1-->4)-C-disaccharides. AB - Starting from methyl 6-bromo-4,6-dideoxy-alpha-D-threo-4-enopyranoside, 4-C branched sugars have been prepared through indium-promoted Barbier-type allylation of various aldehydes in aqueous media followed by hydroboration of the resulting double bond. The intermediate unsaturated monosaccharides were shown to rearrange in acidic media to give 4-C-acetyl-5-C-alkyl pyranose compounds. From beta-1-formyl sugars the corresponding beta-(1-->4)-C-disaccharides were obtained. PMID- 15542084 TI - Synthetic studies on glycosphingolipids from Protostomia phyla: total syntheses of glycosphingolipids from the parasite, Echinococcus multilocularis. AB - Efficient and systematic syntheses of four neutral glycosphingolipids that have been isolated from the metacestodes of Echinococcus multilocularis have been achieved. A key step is the direct glycosylation of galactosyl donors using thioglycosides with benzoyl ceramide in the presence of N-iodosuccinimide (NIS)/TfOH, which gave the desired oligosaccharide derivatives. The fully protected glycosides 13, 20, 22 and 25 were deprotected to give four target glycosphingolipids (1-4). PMID- 15542085 TI - Synthesis of divalent beta-(1-->6)-branched (1-->3)-glucohexaose and trivalent beta-(1-->6)-branched (1-->3)-glucotriose. AB - Hexaose, beta-D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-alpha-D-Glcp-(1-->3)-beta-D Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-beta-D-Glcp, based dimers were synthesized by twofold glycosidation of the hexaosyl trichloroacetimidate with hexylene 1,6 diol, diethylene glycol and triethylene glycol, respectively. Meanwhile, a triose, beta-1D-Glcp-(1-->3)-[beta-D-Glcp-(1-->6)]-beta-D-Glcp, based trimer was obtained by glycosidation of the triosyl trichloroacetimidate with a glycerol derived triol scaffold. PMID- 15542086 TI - Enzymatic glycosidation of sugar oxazolines having a carboxylate group catalyzed by chitinase. AB - Enzymatic glycosidation using sugar oxazolines 1-3 having a carboxylate group as glycosyl donors and compounds 4-6 as glycosyl acceptors was performed by employing a chitinase from Bacillus sp. as catalyst. All the glycosidations proceeded with full control in stereochemistry at the anomeric carbon of the donor and regio-selectivity of the acceptor. The N,N'-diacetyl-6'-O carboxymethylchitobiose oxazoline derivative 1 was effectively glycosidated, under catalysis by the enzyme, with methyl N,N'-diacetyl-beta-chitobioside (4), pent-4-enyl N-acetyl-beta-D-glucosaminide (5), and methyl N-acetyl-beta-D glucosaminide (6), affording in good yields the corresponding oligosaccharide derivatives having 6-O-carboxymethyl group at the nonreducing GlcNAc residue. The N,N'-diacetyl-6-O-carboxymethylchitobiose oxazoline derivative 2 was subjected to catalysis by the enzyme catalysis; however, no glycosidated products were produced through the reactions with 4, 5, and 6. Glycosidation reactions of the beta-d-glucosyluronic-(1-->4)-N-acetyl-D-glucosamine oxazoline derivative 3 proceeded with each of the glycosyl acceptors, giving rise to the corresponding oligosaccharide derivative having a GlcA residue at their nonreducing termini in good yields. PMID- 15542087 TI - In vitro enzymatic modification of puerarin to puerarin glycosides by maltogenic amylase. AB - Puerarin (daidzein 8-C-glucoside), the most abundant isoflavone in Puerariae radix, is prescribed to treat coronary heart disease, cardiac infarction, problems in ocular blood flow, sudden deafness, and alcoholism. However, puerarin cannot be given by injection due to its low solubility in water. To increase its solubility, puerarin was transglycosylated using various enzymes. Bacillus stearothermophilus maltogenic amylase (BSMA) was the most effective transferase used compared with Thermotoga maritima maltosyl transferase (TMMT), Thermus scotoductus 4-alpha-glucanotransferase (TS4alphaGTase), and Bacillus sp. I-5 cyclodextrin glucanotransferase (BSCGTase). TMMT and TS4alphaGTase lacked acceptor specificity for puerarin, which lacks an O-glucoside linkage between D glucose and 7-OH-daidzein. The yield exceeded 70% when reacting 1% puerarin (acceptor), 3.0% soluble starch (donor), and 5U/100 microL BSMA at 55 degrees C for 45 min. The two major transfer products of the BSMA reaction were purified using C(18) and GPC chromatography. Their structures were identified as alpha-d glucosyl-(1-->6)-puerarin and alpha-D-maltosyl-(1-->6)-puerarin using ESI+ TOF MS MS and 13C NMR spectroscopy. The solubility of the transfer products was 14 and 168 times higher than that of puerarin, respectively. PMID- 15542088 TI - Depolymerization of beta-chitin to mono- and disaccharides by the serum fraction from the para rubber tree, Hevea brasiliensis. AB - The serum fraction of latex from Hevea brasiliensis, the para rubber tree, is known to contain an endo-chitinolytic enzyme, hevamine. Herein the activity of the rubber serum towards beta-chitin is investigated. The serum contained 6 mg/mL of protein and a chitinolytic activity of 18 mU permg of protein. The optimum ratio of enzyme to chitin was 0.22 mU/mg, and the optimum substrate concentration was 60 mg/mL. The optimum pH range was pH2-4, and the optimum temperature was 45 degrees C. At these conditions both (GlcNAc)2 and GlcNAc were produced in a molar ratio of approximately 2:1. The hydrolysis of 300 mg of chitin with 64 mU of the rubber serum for 8 days under the optimum conditions gave 39 mg of GlcNAc and 108 mg of (GlcNAc)2 as determined by HPLC. Mixing the rubber serum preparation with an Aspergillus niger pectinase preparation containing beta-N-acetylhexosaminidase can be used to produce almost exclusively the GlcNAc monomer in about 50% yield. PMID- 15542089 TI - Structure and stability of columnar cyclomaltohexaose (alpha-cyclodextrin) hydrate. AB - Rapid recrystallization of cyclomaltohexaose (alpha-cyclodextrin, alpha-CD) from aqueous solution resulted in formation of the columnar crystal structure of alpha CD containing only water as the guest molecule. Complementary water vapor sorption and wide-angle X-ray diffractometry (WAXD) experiments were performed on the alpha-CD columnar structure to elucidate the crystal structure present at varying sorption levels. Equilibrium isothermal water vapor sorption experiments at 40 degrees C revealed that the alpha-CD columnar structure is unstable above a water activity of approximately 0.67. This was confirmed by WAXD diffractograms collected over time, which further revealed that alpha-CD columnar structure undergoes a phase transformation to the cage structure after approximately 0.25 h at 40 degrees C and a water activity of 1.0. PMID- 15542090 TI - Crystal structure of ammonium isosaccharate and aqueous solubility of ammonium and sodium isosaccharates. AB - Ammonium isosaccharate, C6H15NO6.H2O (NH4-ISA), has been synthesized and its crystal structure solved by single-crystal X-ray diffraction methods. NH4-ISA crystallizes in the monoclinic space group P2(1) (#4) with cell parameters a=8.6470(12)A, b=5.0207(7)A, c=9.8193(14)A, beta=91.643(3) degrees , V=426.12(10)A3, Z=2. The structure was refined by full-matrix least-squares on F2 yielding final R-values (all data) R1=0.0485 and Rw2=0.1104. The structure consists of alternating (NH4)+ and (C6H11O6)- layers parallel to the ab plane. An extended network of O-H...O intermolecular (ISA)...(ISA) hydrogen bonds links the (ISA)- anions within the ab plane, while the 3-D connectivity along the c-axis is provided only by (ISA-)...(NH4+)...(ISA-) hydrogen bonds. The aqueous solubility (Si, [ML(-1)]) of NH4- and Na-ISA has been shown to be pH independent at ambient conditions within the range 4.5 or =2500 micro g/plate), both with and without S9-mix. The mutagenic potential of CEDU was further investigated in vivo and in vitro. It did not induce DNA repair in rat hepatocyte primary cultures, and was negative in the micronucleus test in V79 cells and in the comet assay in human leukocytes. In vivo, CEDU was negative in the bone marrow micronucleus test in CD1 mice. The mouse spot test provided a clearly positive result. Treatment of mice on day 9 of pregnancy with 2000 mg/kg resulted in 5.9% of the F1 animals having genetically relevant spots, whereas the corresponding vehicle control group had a spot rate of 1.9%. Since these data clearly identified CEDU as an inducer of gene mutations in vivo, this potential was further investigated in lacZ transgenic Muta Mouse. Six female animals were treated daily on five consecutive days with 2000 mg/kg/day and sacrificed, after a treatment-free sampling time, 14 days later. The data showed a clear increase in the mutant frequency in the bone marrow, the lung and in the spleen. CEDU is an exception in the group of nucleoside analogues, because it was found to be a strong gene mutagen and, in contrast to the other compounds of this group investigated so far, had no considerable clastogenic effects. PMID- 15542108 TI - Induction of A:T to G:C transition mutations by 5-(2-chloroethyl)-2'-deoxyuridine (CEDU), an antiviral pyrimidine nucleoside analogue, in the bone marrow of Muta Mouse. AB - 5-(2-chloroethyl)-2'-deoxyuridine (CEDU) is a pyrimidine nucleoside analogue formerly in development for the treatment of herpes simplex virus infections. The compound proved clearly mutagenic in the mouse spot test and exhibited weak activity in the Salmonella reverse mutation test, which led to the termination of the compound's development. In another study, CEDU, administered orally to beta galactosidase (lacZ) transgenic mice (Muta Mouse) for five days, induced a clear increase in lacZ mutant frequencies in spleen, lung, and bone marrow. In the present follow-up study, we analyzed 32 of those lacZ mutants isolated from the bone marrow of the Muta Mouse animals of the experiments mentioned above, in order to obtain further information on the type of mutations induced by CEDU. CEDU induced a pronounced increase in A:T to G:C transitions. The distribution of A:T to G:C transitions was clearly non-random, showing a bias towards T to C substitutions in the coding DNA strand and a preference to occur in the sequence motif 5'-(G or C)-T-G-3'. Our data support the hypothesis that CEDU, after being phosphorylated, is incorporated into cellular DNA in place of thymidine, which leads to mispairing with guanosine during subsequent DNA replication. As a result, the compound is thought to exert its mutagenicity by inducing mismatches leading to T to C transitions. Our findings point towards a mode of mutagenic action of CEDU that differs fundamentally from that of other antiviral antinucleosides whose clastogenic and recombinogenic activities prevail. PMID- 15542109 TI - A comparative investigation of DNA strand breaks, sister chromatid exchanges and K-ras gene mutations induced by cadmium salts in cultured human cells. AB - Cadmium (Cd) is a toxic heavy metal of continuing occupational and environmental concern with a wide variety of adverse effects. Several studies have shown that cadmium produces DNA strand breaks, DNA-protein cross-links, oxidative DNA damage, chromosomal aberrations, dysregulation of gene expression resulting in enhanced proliferation, depressed apoptosis and/or altered DNA repair. This study was undertaken to investigate the ability of cadmium chloride (CdCl(2)) and cadmium sulphate (CdSO(4)) to induce point mutations in codon 12 of the K-ras protooncogene assessed by polymerase chain reaction-single strand conformation polymorphisms (PCR-SSCP) and RFLP-enriched PCR methods. Also their genotoxic effects were analyzed by the comet assay and sister chromatid exchanges test. The human lung fibroblast cell line MRC-5 was used for the experiments. Sister chromatid exchanges assay (SCEs) frequencies were significantly increased in cells exposed to cadmium salts in relation to controls (p<0.001). Despite the slow increment observed in the three comet parameters considered when cells were treated with cadmium chloride, significant differences between groups were only found in the variable comet moment (CM) (p<0.005). On the other hand, when cells were exposed to cadmium sulphate, the Kruskal-Wallis test showed highly significant differences between groups for migration, tail moment and comet moment parameters (p<0.001). Nevertheless, a null or weak point mutation induction in K-ras protooncogene was detected using polymerase chain reaction-low ionic strength-single strand conformation polymorphisms (PCR-LIS-SSCP) and RFLP enriched PCR methods when cells were treated with cadmium salts. Thus, inorganic cadmium produces genotoxicity in human lung fibroblast MRC-5 cells, in the absence of significant point mutation of the K-ras gene. PMID- 15542110 TI - Microsatellite mutation directed by an external stimulus. AB - Microsatellites are regions of DNA containing tandem repeats of a core 2-6 bp nucleotide sequence. To test the hypothesis that microsatellite mutation can be directed by exposure to specific external cues, control and treatment groups of resistant and susceptible wheat varieties were grown under controlled conditions and genotyped at a number of microsatellite loci that map to chromosomes known to contain Fusarium head blight (FHB) resistance/susceptibility loci. Genotyping was undertaken both prior to and following exposure to Fusarium graminearum, the FHB pathogen. Within a month of inoculation of inflorescences, 58% of experimental plants, and no control plants, had acquired a novel allele at the locus Xgwm112.1. This allele was detected only in head blight affected tissue. Uninoculated control plants, and leaf samples from inoculated plants, showed no mutation. Cloning and sequencing of PCR products indicates that the new allele was generated by contraction of the (CT)(n) repeat motif. Observation of the same deletion-based mutation in all varieties, its absence in control plants not exposed to the head blight pathogen, and the detection of no similar mutational events in a control panel of loci not expected to show mutation, indicates that this example of microsatellite mutation is induced and/or caused by FHB infection. PMID- 15542111 TI - DNA damage and mutations produced by chloroacetaldehyde in a CpG-methylated target gene. AB - Chloroacetaldehyde (CAA) is a metabolite of the human carcinogen vinyl chloride. CAA produces several types of DNA adducts including the exocyclic base adducts 3,N(4)-ethenocytosine, 1,N(6)-ethenoadenine, N(2),3-ethenoguanine, and 1,N(2) ethenoguanine. Adducts of CAA with 5-methylcytosine have not yet been characterized. Here we have analyzed the mutational spectra produced by CAA in the supF gene of the pSP189 shuttle vector when present in either an unmethylated or CpG-methylated state. The vectors were replicated in human nucleotide excision repair-deficient XP-A fibroblasts. The mutational spectra obtained with the unmethylated and methylated supF target genes were generally similar with a preponderance of C/G to T/A transitions and C/G to A/T transversions. CAA-induced DNA adducts were mapped along the supF gene by using thermostable thymine DNA glycosylase (TDG) in conjunction with ligation-mediated PCR or by a Taq polymerase stop assay. Prominent CAA-induced TDG-sensitive sites were seen at several CpG positions but were independent of methylation. Methylated CpG sites were sites of CAA-induced mutations but were not the major mutational hotspots. Taq polymerase arrest sites were observed at numerous sequence positions in the supF gene and reflected the rather broad distributions of mutations along the sequence. We conclude that methylated CpG sites are not preferential targets for chloroacetaldehyde-induced mutagenesis. PMID- 15542112 TI - From mutation to malignancy. PMID- 15542113 TI - Information dynamics in carcinogenesis and tumor growth. AB - The storage and transmission of information is vital to the function of normal and transformed cells. We use methods from information theory and Monte Carlo theory to analyze the role of information in carcinogenesis. Our analysis demonstrates that, during somatic evolution of the malignant phenotype, the accumulation of genomic mutations degrades intracellular information. However, the degradation is constrained by the Darwinian somatic ecology in which mutant clones proliferate only when the mutation confers a selective growth advantage. In that environment, genes that normally decrease cellular proliferation, such as tumor suppressor or differentiation genes, suffer maximum information degradation. Conversely, those that increase proliferation, such as oncogenes, are conserved or exhibit only gain of function mutations. These constraints shield most cellular populations from catastrophic mutator-induced loss of the transmembrane entropy gradient and, therefore, cell death. The dynamics of constrained information degradation during carcinogenesis cause the tumor genome to asymptotically approach a minimum information state that is manifested clinically as dedifferentiation and unconstrained proliferation. Extreme physical information (EPI) theory demonstrates that altered information flow from cancer cells to their environment will manifest in-vivo as power law tumor growth with an exponent of size 1.62. This prediction is based only on the assumption that tumor cells are at an absolute information minimum and are capable of "free field" growth that is, they are unconstrained by external biological parameters. The prediction agrees remarkably well with several studies demonstrating power law growth in small human breast cancers with an exponent of 1.72+/-0.24. This successful derivation of an analytic expression for cancer growth from EPI alone supports the conceptual model that carcinogenesis is a process of constrained information degradation and that malignant cells are minimum information systems. EPI theory also predicts that the estimated age of a clinically observed tumor is subject to a root-mean square error of about 30%. This is due to information loss and tissue disorganization and probably manifests as a randomly variable lag phase in the growth pattern that has been observed experimentally. This difference between tumor size and age may impose a fundamental limit on the efficacy of screening based on early detection of small tumors. Independent of the EPI analysis, Monte Carlo methods are applied to predict statistical tumor growth due to perturbed information flow from the environment into transformed cells. A "simplest" Monte Carlo model is suggested by the findings in the EPI approach that tumor growth arises out of a minimally complex mechanism. The outputs of large numbers of simulations show that (a) about 40% of the populations do not survive the first two-generations due to mutations in critical gene segments; but (b) those that do survive will experience power law growth identical to the predicted rate obtained from the independent EPI approach. The agreement between these two very different approaches to the problem strongly supports the idea that tumor cells regress to a state of minimum information during carcinogenesis, and that information dynamics are integrally related to tumor development and growth. PMID- 15542114 TI - Clinicopathologic features in colorectal cancer patients with microsatellite instability. AB - The microsatellite instability (MSI) mutational pathway is critical to carcinogenesis in a small but significant proportion of colorectal cancers. While MSI is identified in most cancers in individuals with hereditary non-polyposis colorectal cancer, the majority of MSI tumors are found in individuals with sporadic disease. Colorectal cancers arising as a result of MSI have distinct clinicopathologic features distinguishing them from those with microsatellite stability. MSI colorectal cancers affect a larger percentage of women, are usually localized proximal to the splenic flexure, and have a higher incidence of synchronous and metachronous tumors. They are associated with a mucinous histology, tumor-infiltrating lymphocytes, a Crohn's-like inflammatory response, and a higher grade but lower stage. Overall survival is better in individuals with MSI. The benefit of chemotherapy in MSI colorectal cancers, with and without lymph node metastases, remains unclear. PMID- 15542115 TI - Genomic heterogeneity and instability in colorectal cancer: spectral karyotyping, glutathione transferase-Ml and ras. AB - Genomic instability in cancer is frequently described as being either chromosomal instability or microsatellite instability, although when events within chromosomes are monitored, extensive intrachromosomal instability is also found. Spectral karyotyping was used to visualize how extensively genomic instability gives rise to intratumor genomic heterogeneity in sporadic colorectal carcinomas. Two factors were then examined which might relate to intrachromosomal instability in colorectal cancers: the presence of the glutathione transferase-Ml gene to detoxify potential carcinogens, and the presence of activated ras which has been associated with chromosomal instability when first expressed. Intrachromosomal genomic instability was previously determined by inter-(simple sequence repeat) PCR (inter-SSR PCR) and by fractional allelic loss rate for 348 markers. GSTM1 status was determined for each of 49 tumors through use of specific PCR, and 28 of the tumors showed the GSTM1 null genotype. A significant association was found between GSTMl-null status and elevated inter-(simple sequence repeat) PCR instability. In contrast, no association was found with fractional allelic loss rate. The first exons of the K-ras and H-ras oncogenes were sequenced in 72 colorectal cancers; 19 of the tumors had a mutation in codon 12 of the K-ras gene (24.5%), but no H-ras mutations were found. A weak correlation (p=0.10) was observed between mutant K-ras and inter-(simple sequence repeat) PCR genomic instability, and no association existed with fractional allelic loss rate. PMID- 15542117 TI - The genesis and evolution of acrylic bone cement. AB - In 1957 John Chamley and I began to discuss the question of cement fixation of femoral prostheses. This was an independent development, because we had no knowledge of any other work in this area. Our conclusion was to grout in the prosthesis with a suitable cement. I selected a cold-curing acrylic denture repair material called Nu-Life as most appropriate. This material proved satisfactory in laboratory and clinical trials, and in 1958 Chamley undertook hip replacement using a cemented femoral prosthesis in six patients. This clinical success aroused great interest and some skepticism, because it was not always appreciated that Charnley and I had a substantial body of scientific data that justified this new approach to arthroplasty. Not surprisingly,the clinical aspects received most emphasis in the next decade. The scientific data were published mostly in the dental literature and were forgotten by the time cementation arthroplasty was generally accepted and so were rediscovered in subsequent years. PMID- 15542118 TI - Charnley low frictional torque arthroplasty: clinical developments. PMID- 15542119 TI - Acrylic bone cements: composition and properties. AB - Cementing endoprostheses is one of the most frequently performed orthopedic procedures in the world today. The bone cements that are currently available commercially are all based on the chemical substance methylmethacrylate. Although all of them have the same chemical basis, acrylic bone cements are not all alike. This article discusses the composition of acrylic bone cements and the influence of the composition on the properties of the material. PMID- 15542120 TI - Acrylic bone cements: mechanical and physical properties. AB - Mechanical and physical properties are of particular significance for the performance of acrylic bone cement. Several mechanical test methods are described in the literature to characterize the mechanical performance of bone cements. The simulation of the in vivo situation is extremely difficult, however, because of the complex mechanism of loading in the bone. The usefulness of the different mechanical and physical test methods, several results of commercial acrylic bone cements, and the influence of different parameters, such as temperature, test environment, and preparation of specimens on these results are discussed in this article. PMID- 15542121 TI - Wear particulate and osteolysis. AB - Total joint replacements of the hip and knee are generally highly successful, with satisfactory longevity and clinical results. Using modern biocompatible materials, optimal component design, and meticulous surgical technique, survivorship of cemented or cementless joint replacements is approximately 15 years with more than a 90% probability. The host's biologic response is critical to implant longevity. Particulate disease refers to the host's adverse biologic response to wear debris and byproducts generated from the prosthesis. Initially, emphasis was placed on particulate polymethylmethacrylate (cement disease), but more recently polyethylene wear debris has been underscored. Debris from several materials in sufficient quantities and physicochemical forms, however, can generate an inflammatory cascade resulting in periprosthetic bone destruction (osteolysis), jeopardizing long-term success of the implant. PMID- 15542122 TI - The role of polymethylmethacrylate bone cement in revision arthroplasty of the hip. AB - When restoration of bone stock is necessary in revision arthroplasty of the hip using morselized or structural allograft bone, cement is used. Cement strengthens allograft bone, impairs its resorption, allows anatomic positioning of components (ie, cementing the cup in a protective ring), and allows delivery of antibiotics. Allograft bone provides a perfect lattice for cementation and also allows a cementless reconstruction as far as host bone is concerned. PMID- 15542123 TI - Acrylic bone cements: clinical developments and current status: Scandinavia. AB - This article focuses on bone cement, the cementing technique used, and their influence on aseptic loosening and infection of acrylic and joint implants- Scandinavian view. PMID- 15542124 TI - Clinical development and current status: Europe. AB - This article reviews the development and current status of cemented fixation in total hip replacement in Europe. Key points include the wide country-to-country variation in use of cemented vs. non-cemented fixation and the largely overlooked importance of the choice of bone cement as a factor highly correlated with clinical outcome. Laboratory studies by the authors are also reviewed. Results suggest that the type of acrylic bone cement used affects wear phenomena at the implant/cement interface. Further studies by microcalorimetry suggest that certain aspects of acrylic starting materials (low molecular weight and use of radiation sterilization) affect long-term physico-chemical stability and may thus influence clinical outcomes. PMID- 15542125 TI - Acrylic bone cement: clinical development and current status in North America. AB - This article discusses issues surrounding acrylic bone cement and covers functional results, mechanism and definition of failure, design issues, wear and alternate bearing surfaces, revision, and evolution of concepts regarding infection, dislocation, thromboembolic issues, surgical approach, bone loss and bone grafts, and finally a couple of problems the author sees for the future. PMID- 15542126 TI - Acrylic bone cement: its clinical development and current status in Japan. AB - This article describes the transition of annual total sales figures of bone cement in Japan during the last 12 years, bone cement-related complications, the present status of bone cement application in Japan as surveyed by the Implant Committee of the Japanese Orthopaedic Association, and its proposed precautions to be taken for the prevention of bone cement complications. PMID- 15542127 TI - Injectable calcium phosphate cement for bone repair and implant fixation. AB - The studies as described are aimed at determining the efficacy of newly developed calcium phosphate cement when this material is used as a bone defect filler or gap filler around metal implants. An overview is provided about bone graft substitutes and methods of metal implant fixation. PMID- 15542128 TI - The potential of tissue engineering in orthopedics. AB - This article presents models of human phalanges and small joints developed by tissue engineering. Biodegradable polymer scaffolds support growth of osteoblasts, chondrocytes, and tenocytes after implantation of the models in athymic mice. The cell-polymer constructs are vascularized by the host mice, form new bone, cartilage, and tendon with characteristic gene expression and protein synthesis and secretion, and maintain the shape of human phalanges with joints. The study demonstrates critical progress in the design and fabrication of bone, cartilage, and tendon by tissue engineering and the potential of this field for human clinical orthopedic applications. PMID- 15542129 TI - Biologic fixation and bone ingrowth. AB - Total hip arthroplasty has provided thousands of patients with pain relief and has improved their quality of life. Advances in orthopaedic surgical techniques and implant biomaterials now allow predictable surgical results in most patients. Despite the overwhelming success of this surgical procedure, the debate continues surrounding the optimal choice of implants. Femoral and acetabular implants with varying geometries and fixation methods are currently available. Acrylic bone cement has been used extensively in the past for acetabular and femoral fixation. This mode of component fixation currently remains the technique used most frequently throughout Europe and has shown excellent long-term results. Problems inherent with acrylic bone cement, however, have encouraged other surgeons to use alternative surfaces to allow biologic fixation. PMID- 15542130 TI - Cementless implant fixation--toward improved reliability. AB - Cementless implants offer the advantage of fixation by direct bone-to-implant osseointegration, thereby avoiding the use of a synthetic intermediary material (such as acrylic bone cement) of limited mechanical strength. Successful osseointegration, however, depends on several conditions being satisfied during the peri-implant bone healing period, including the need for limited early loading resulting in minimal relative movement at the implant-bone interface. Sintered porous- and plasma spray-coated implants represent the most common cementless orthopedic implants in current clinical use, although novel cast structures also are being investigated. All stand to benefit from surface modifications currently being explored to enhance osteoconductive or osteoinductive characteristics of the implants. The faster osseointegration that such modified surface designs potentially might offer would result in more reliable and convenient (from the patient perspective) cementless implants. Encouraging results of early animal-based studies exploring such modifications have been reported. PMID- 15542132 TI - Medicare reform and your practice: what you need to know. AB - Reimbursements for Medicare Part B physician-administered medications have been cut and will be further reduced in the years to come. Physician groups have supported the prescription benefit within the Medicare reform package. However, physician and patient advocacy groups alike are concerned about how other portions of the legislation will change the administration of Medicare Part B covered drugs in the office setting. This article explores the payment changes built into the Medicare Prescription Drug, Improvement, and Modernization Act of 2003, what the proponents and opposition have been saying, and how these changes will impact the practice of urology and patients with prostate cancer. PMID- 15542133 TI - Practice management and patient care: is there an ideal situation for both patients and physician practices in light of legislative reform? AB - The Medicare reimbursement rules have changed, and urologists must be prepared to change as well. They must take an active role in the management of their office practices, for the alternative will almost certainly have a negative impact on access for many patients, including those with prostate cancer. Urologists need to critically evaluate the daily operation of their practice, with an eye toward eliminating excess and increasing efficiency. This article discusses areas, such as staffing, collections, coding, and ancillary services, which can be optimized to meet the requirements of a successful urology practice in this changing environment. PMID- 15542136 TI - Optimized large-scale production of high titer lentivirus vector pseudotypes. AB - The goal of the present study was to develop an efficient transient transfection method for large-scale production of high titer lentivirus vector stocks of eight different pseudotypes. The envelope genes used for this purpose were those from VSV-G, Mokola, Rabies, MLV-Ampho, MLV-10A1, LCMV-WE, and LCMV-Arm53b. All envelopes were cloned into phCMV, which yielded lentivirus vector titers one, two, or three orders of magnitude higher than the original plasmids for the Rabies, MLV-10A1, and MLV-Ampho envelopes, respectively. When these newly constructed envelope expression plasmids were used for packaging, treatment with sodium butyrate resulted in almost five-fold increase in titers for some of the pseudotypes, had no effect for others (VSV-G and Rabies), and negatively impacted titers for the LCMV-derived pseudotypes. Production of vectors in serum-free media yielded titers only slightly lower than those obtained in the presence of serum. The efficiency of concentrating vector supernatants by ultracentrifugation or ultrafiltration was compared, with higher recovery efficiencies for the latter method, but the highest titers for most pseudotypes were obtained by ultracentrifugation. The best conditions for each individual pseudotype yielded lentivirus vector stocks with titers above 1 x 10(9) tu/mL for most pseudotypes, and higher than 1 x 10(10) tu/mL for VSV-G. PMID- 15542137 TI - A natural vaccinia virus promoter with exceptional capacity to direct protein synthesis. AB - A survey of vaccinia virus promoters, through a reporter gene approach, has identified the viral I1L promoter as having exceptional activity. The I1L promoter exhibited over 10 times the activity of other vaccinia promoters and even rivaled the activity of the bacteriophage T7 promoter in the hybrid vaccinia/T7 expression system. The I1L promoter had high activity in both transient transfection experiments and in the context of recombinant viruses. The I1L promoter should be useful for high-level protein synthesis and poxvirus studies in general. PMID- 15542138 TI - Effects of freezing and storage temperature on MS2 viability. AB - Monitoring human enteric virus levels in domestic wastewater effluent is crucial to protecting human health. Occasionally, during intensive sampling, wastewater samples must be stored for later viral analysis. Little data exist regarding how enteric viruses survive during storage at different temperatures in secondary treated wastewater. During a field-scale study assessing pathogen removal performance by various onsite treatment technologies, the MS2 bacteriophage, an indicator of enteric viruses, was inoculated into septic tank (STE), sand filter, peat filter and constructed wetland (CW) effluents to determine virus decay at various storage temperatures. Virus stored at temperatures > or =10 degrees C and at -20 degrees C decayed nearly twice as fast as those stored at 4 degrees C or 80 degrees C. Decreased water quality decreased viral decay rates at 4 degrees C and -80 degrees C, with slowest decay occurring in STE and the fastest in sterile PBS and low pH peat effluent. In CW effluent after 8 days, less MS2 was inactivated when stored at 4 degrees C (20%) compared to -80 degrees C (58%); however, during extended storage (approximately 300 days), less MS2 was inactivated at -80 degrees C (75%) compared to 4 degrees C (93%). We recommend that viruses in wastewater be stored in the dark at 4 degrees C unless storage for >40 days is necessary. PMID- 15542139 TI - Biolistic inoculation of plants with viroid nucleic acids. AB - Parameters for biolistic transfer of viroid nucleic acids using a Helios Gene Gun device were assayed. The main achievement of this method is high efficiency of inoculation with linear monomeric viroid cDNAs and RNAs. This greatly facilitates the study of mutated sequence variants, viroid libraries and mixed populations. The lower limits for efficient inoculation of monomeric cDNA fragments with the sequence of potato spindle tuber viroid (PSTVd) and native PSTVd RNA as detected 21 days p.i. are in the range of 50 ng and 200 pg per tomato plant, respectively. At a higher dose, i.e. 2 ng of native RNA per plant, biolistic transfer causes drastic stunting compared to conventional mechanical inoculation, which points to higher PSTVd titers after the biolistic transfer. Infection is readily achieved with exact length monomeric RNA transcripts having 5'-triphosphate and 3'-OH termini in amounts ranging from 2 to 20 ng per plant, suggesting no need for any supplementary modifications of ends or RNA circularization. The biolistic transfer is efficient for viroid "thermomutants", which exhibit low or no infectivity with conventional mechanical inoculation with Carborundum. The biolistic inoculation is also efficient for two other members of the Pospiviroidae family, hop stunt and hop latent viroid. PMID- 15542140 TI - Apoptosis induced by bovine ephemeral fever virus. AB - The potential significance of bovine ephemeral fever virus (BEFV)-induced apoptosis and involved viral molecules was fully unknown. In the present study, evidence is provided demonstrating that bovine ephemeral fever virus induces apoptosis in several cell lines. Five types of assays for apoptosis were used in examining BEFV-infected cells. (1) Assay for DNA fragmentation, (2) nuclear staining with acridine orange, (3) ELISA detection of cytoplasmic histone associated DNA fragment, (4) terminal deoxynucleotidyltransferase-mediated dUTP biotin nick-end labelling (TUNEL) assay of BEFV-infected cells, (5) observation of blebbing of the plasma membrane and the formation of apoptotic bodies of apoptic cells by scanning electron microscope. The level of lactate dehydrogenase (LDH) in BEFV-infected cells was increased significantly after 20-25 h post infection. Caspases-2, -3, -4, -6, -8, -9, and -10 were activated in BEFV infected BHK-21 cells. To determine further whether BEFV-induced apoptosis was caspase-dependent, the effect of the tripeptide pan-ICE (caspase) inhibitor N benzyloxycarbonyl-Val-Ala-Asp-fluoromethyketone on the inhibition of apoptosis in BEFV-infected BHK-21 cells, was investigated. Apoptosis could be blocked by the caspase inhibitor (Z-VAD-fmk), indicating that BEFV induces caspase-dependent apoptosis in cultured cells. PMID- 15542141 TI - Quantitation of porcine circovirus type 2 isolated from serum/plasma and tissue samples of healthy pigs and pigs with postweaning multisystemic wasting syndrome using a TaqMan-based real-time PCR. AB - Porcine circovirus type 2 (PCV2) has been linked to several disease syndromes during the last decade. In this context, postweaning multisystemic wasting syndrome (PMWS) has emerged as a significant disease. As most pig herds are infected with PCV2, the determination of viral load in animals may be useful in discriminating between healthy and PMWS pigs. A TaqMan-based real-time PCR for quantitation of PCV2 in serum/plasma and tissue samples was established. A standard curve was created from serial dilutions of a plasmid encoding ORF2 (cap gene) of PCV2 and used to estimate the number of viral DNA copies in the analyzed samples. Comparison of viral load in mesenteric lymph nodes and serum/plasma from healthy animals and PMWS animals showed statistical significant difference between the two groups (p < 0.01). No healthy pigs had viral load greater than 10(6) PCV2 genomes per ml serum or 500 ng tissue sample, while all clinically sick PMWS pigs had PCV2 loads above 10(7) in both serum/plasma and in tissue samples. Furthermore, the estimated viral load in tissue samples from PMWS pigs was related to the immunohistochemical findings, with especially lymph nodes, ileum, and tonsil giving both high viral load, and a high degree of staining by immunohistochemistry. PMID- 15542142 TI - Detection of HCV antibodies in oral fluid. AB - Although conventionally the detection of HCV antibodies is carried out on serum, the collection of oral fluid is non-invasive, safe and cost effective. In this study, the efficacy of the detection of HCV antibodies in oral fluid was assessed. 73 anti-HCV positive and 73 anti-HCV negative paired serum/oral fluid samples, drawn from patients visiting a Belgian academic hospital, were tested using the modified Ortho HCV 3.0 and LIA confirmation assay. Performing the test on oral fluid with the modified protocol, 61/73 anti-HCV positive samples were tested positive, while 73/73 anti-HCV negative samples were tested negative, giving a sensitivity and specificity of 83.6% (95% CI: 72.7-90.9%) and 100.0% (95% CI: 93.8-100.0%), respectively. Comparing S/CO of concordantly positive and negative samples, the cut-off point was lowered by 30% resulting in a sensitivity of 89.0% (95% CI: 79.0-94.8%) while the specificity remained 100.0% (95% CI: 93.8 100.0%). The confirmation assay was carried out as described by the manufacturer, diluting the oral fluid 1:10. Testing paired samples gave a concordance of 85.6% (125/146), yielding no more accurate results. These findings suggested that the modified ELISA method for anti-HCV detection in oral fluid can be used for epidemiological surveys. PMID- 15542143 TI - Evaluation of the sensitivity and specificity of six HIV combined p24 antigen and antibody assays. AB - In this study, we evaluated the performance of six HIV combined p24 antigen and antibody (Ag/Ab) assays versus two third-generation anti-HIV antibody assays. The assays were evaluated using p24 antigen panel of 31 HIV-1 subtypes (n = 124), 25 HIV-1 seroconversion panels (n = 176), HIV-1 antibody positive samples including group M subtypes and group O (n = 559), HIV-2 positive samples (n = 110), and unselected HIV negative samples from four French private laboratories (n = 1005). The results showed that overall HIV combined Ag/Ab assays present better performance, when compared to antibody-only assays. However, some differences were observed in the sensitivity of the six HIV combined Ag/Ab assays evaluated. The AxSYM and Murex Combo assays had the best sensitivity score in this study and reduced the window period by 2.0-2.35 days relative to antibody only assays and 1 2.17 days relative to the other combined Ag/Ab assays. Among combined HIV Ag/Ab assays, Genscreen Plus and AxSYM Combo presented the highest specificity, with 99.9% and 99.8%, respectively. PMID- 15542144 TI - Efficient purification of bromoviruses by ultrafiltration. AB - Ultrafiltration using polyethersulfone-membranes was evaluated as an efficient and preferred method for purifying Cowpea Chlorotic Mottle Virus (CCMV). Cesium chloride (CsCl) ultracentrifugation and ultrafiltration protocols are described, and comparative UV-spectroscopic and electron micrograph results are presented. CCMV purified by ultrafiltration are shown to be equivalent to CCMV purified by ultracentrifugation, while reducing purification time by two days and avoiding the need for expensive capital overheads such as ultracentrifuges, rotors and toxic CsCl chemical waste. PMID- 15542145 TI - Semiconductor quantum dots as contrast agents for whole animal imaging. AB - Recent developments in quantum dot (QD) technology have paved the way for using QDs as optical contrast agents for in vivo imaging. Pioneering papers showed the use of QDs as luminescent contrast agents for imaging cancer and guiding cancer surgery. The possible future use of QDs for clinical applications is expected to have a significant impact, however many challenges in this field have yet to be overcome. PMID- 15542146 TI - The maturing of the human embryonic stem cell transcriptome profile. AB - Several recent reports used microarray, serial analysis of gene expression (SAGE), and expressed sequence tags (EST) strategies to characterize the human embryonic stem cell transcriptome and those of their differentiated derivatives. All three approaches yielded valuable data and highlight the fact that a large percentage of genes remain uncharacterized in these cells representative of early human development. PMID- 15542147 TI - The search for new beta-cell sources. AB - Type 1 diabetes affects an estimated 150 million people worldwide and results from an autoimmune-mediated destruction of insulin-producing beta-cells. In the United States alone more than 16 million people are affected by this disease and it is estimated that spending for diabetes management accounts for one out of every eight healthcare dollars. In this context, scientists are proposing novel therapeutic strategies that might allow a perfect glycemic control of most patients with diabetes. PMID- 15542148 TI - New hepatocytes for toxicology? AB - In a recent article M.G. Sacco and co-workers described the establishment of immortalized untransformed transgenic hepatocyte (MMH-GH) cell lines, obtained from a cross between the AT/cytoMet and Hsp70/hGH transgenic mice. This strategy proved to be successful because the MMH-GH showed stability in culture and sensitivity to chemical exposure. Based on these results, the MMH-GH cell lines could prove to be a valid alternative cell-based assay for use in toxicological studies. PMID- 15542150 TI - Peptide nucleic acids on microarrays and other biosensors. AB - The analysis of biomolecules using microarrays and other biosensors has a significant role in molecular biotechnology, and will become even more important in the future as a versatile tool for research and diagnostics. For many applications, the synthetic DNA mimic peptide nucleic acid (PNA) could be advantageous as a probe molecule, owing to its unique physicochemical and biochemical properties. PNA exhibits superior hybridization characteristics and improved chemical and enzymatic stability relative to nucleic acids. Furthermore, its different molecular structure enables new modes of detection, especially procedures that avoid the introduction of a label. In our opinion, all of these factors contribute significantly toward the establishment of faster and more reliable analytical processes and opens new fields of application. PMID- 15542151 TI - Gene therapy meets vaccine development. AB - Therapeutic vaccines such as those used to combat cancer or persistent viral infection are required to reprogramme a downregulated immune system. This presents a difficult challenge for vaccine design and merits the development of novel immunization protocols. Currently, we know that mobilization of dendritic cells (DCs) to present antigens to T lymphocytes is crucial for effective immunization. Our increasing understanding of DC biology, coupled with the growing sophistication of viral vectors developed for gene therapy, makes more rational vaccine design an exciting possibility. Here we propose that engineering viral vectors to express antigens in activated DCs will provide the most effective vaccines for priming an immune response. PMID- 15542152 TI - Marker-free site-specific gene integration in plants. AB - For nearly 15 years, the use of site-specific recombination systems in plants has focused on the deletion or integration of DNA. Each of these applications offers practical solutions to two problems in biotechnology: the presence of unneeded DNA in the transgene locus and variation in the locus structure among independent transgenic lines. Given that each of these separate applications is becoming more practical for commercial use, it is time to consider combining them into an integrated technology. Here we propose a strategy for a "combined step" method that makes use of two site-specific recombination systems: one for integrating the DNA and a second for removing sequences that are not needed after DNA transfer. This strategy is based on published data and exemplifies the use of the Cre-lox, FLP-FRT and R-RS inducible systems. PMID- 15542153 TI - Microarrays--status and prospects. AB - Microarrays have become an extremely important research tool for life science researchers and are also beginning to be used in diagnostic, treatment and monitoring applications. This article provides a detailed description of microarrays prepared by in situ synthesis, deposition using microspotting methods, nonplanar bead arrays, flow-through microarrays, optical fiber bundle arrays and nanobarcodes. The problems and challenges in the development of microarrays, development of standards and diagnostic microarrays are described. Tables summarizing the vendor list of various derivatized microarray surfaces, commercially sold premade microarrays, bead arrays and unique microarray products in development are also included. PMID- 15542154 TI - Patenting for the research scientist: an update. AB - Academic institutional research constantly produces results worthy of patent protection, but coping with the demands of patent law presents considerable challenges to bioscientists working in these institutions. Inventors need, however, to be aware of recent patent office guidelines and court decisions if they are to seek useful intellectual property as a basis for technology transfer to industry. PMID- 15542155 TI - Rapid prototyping in tissue engineering: challenges and potential. AB - Tissue engineering aims to produce patient-specific biological substitutes in an attempt to circumvent the limitations of existing clinical treatments for damaged tissue or organs. The main regenerative tissue engineering approach involves transplantation of cells onto scaffolds. The scaffold attempts to mimic the function of the natural extracellular matrix, providing a temporary template for the growth of target tissues. Scaffolds should have suitable architecture and strength to serve their intended function. This paper presents a comprehensive review of the fabrication methods, including conventional, mainly manual, techniques and advanced processing methods such as rapid prototyping (RP) techniques. The potential and challenges of scaffold-based technology are discussed from the perspective of RP technology. PMID- 15542156 TI - Impact of genomics on microbial food safety. AB - Genome sequences are now available for many of the microbes that cause food-borne diseases. The information contained in pathogen genome sequences, together with the development of themed and whole-genome DNA microarrays and improved proteomics techniques, might provide tools for the rapid detection and identification of such organisms, for assessing their biological diversity and for understanding their ability to respond to stress. The genomic information also provides insight into the metabolic capacity and versatility of microbes; for example, specific metabolic pathways might contribute to the growth and survival of pathogens in a range of niches, such as food-processing environments and the human host. New concepts are emerging about how pathogens function, both within foods and in interactions with the host. The future should bring the first practical benefits of genome sequencing to the field of microbial food safety, including strategies and tools for the identification and control of emerging pathogens. PMID- 15542157 TI - Multi-modality treatment in oesophageal cancer: a curse for radiotherapy progress. PMID- 15542158 TI - The radiation-induced fibroatrophic process: therapeutic perspective via the antioxidant pathway. AB - The radiation-induced fibroatrophic process (RIF) constitutes a late, local and unavoidable sequela to high-dose radiotherapy, traditionally considered irreversible. Today, this process is partly reversible, thanks to recent progress in understanding the physiopathology of the lesions it causes and the results of recent clinical trials using antioxidant therapy. This review includes a synthetic description of the static and dynamic features of the RIF process, as reflected by its clinical, instrumental and histopathological characteristics, and by its cellular and molecular regulation. Schematically, three successive clinical and histopathological phases can be distinguished: a pre-fibrotic aspecific inflammatory phase, a constitutive fibrotic cellular phase, and a matrix densification and remodelling phase, possibly ending in terminal tissular necrosis. The respective roles of the chief actors in the RIF process are defined, as well as their development with time. A fibroblastic stromal hypothesis is suggested revolving around a 'gravitational effect' exerted by the couple ROS (reactive oxygen species)--fibroblasts, and partly mediated by TGF beta1. A variety of strategies have been tested for the management of RIF. In the light of the mechanisms described, a curative procedure has been proposed via the antioxidant pathway. In particular, it was showed that superoxide dismutase and combined pentoxifylline-tocopherol treatment enables the process of established radiation-induced fibroatrophy to be greatly reduced or even reversed, both in clinical practice and animal experiments. The efficacy of combined pentoxifylline tocopherol treatment in superficial RIF was confirmed in a randomised clinical trial, and then in successful phase II trials especially in uterine fibroatrophy and osteoradionecrosis. It is of critical importance to evaluate these new management approaches in larger clinical trials and to improve the recording of results for better outcome analysis. Mechanistic studies are always necessary to improve understanding of the RIF process and the antifibrotic drug action. PMID- 15542159 TI - Double-blind placebo-controlled randomised trial of vitamin E and pentoxifylline in patients with chronic arm lymphoedema and fibrosis after surgery and radiotherapy for breast cancer. AB - BACKGROUND AND PURPOSE: Treatment-induced arm lymphoedema is a common and distressing complication of curative surgery and radiotherapy for early breast cancer. A number of studies testing alpha-tocopherol (vitamin E) and pentoxifylline suggest evidence of clinical regression of superficial radiation induced fibrosis but there is only very limited evidence from randomised trials. Arm lymphoedema after lymphatic radiotherapy and surgery has been used in the present study as a clinical system for testing these drugs in a double-blind placebo-controlled randomised phase II trial. PATIENTS AND METHODS: Sixty-eight eligible research volunteers with a minimum 20% increase in arm volume at a median 15.5 years (range 2-41) after axillary/supraclavicular radiotherapy (plus axillary surgery in 51/68 (75%) cases) were randomised to active drugs or placebo. All volunteers were given dl-alpha tocopheryl acetate 500 mg twice a day orally plus pentoxifylline 400 mg twice a day orally, or corresponding placebos, for 6 months. The primary endpoint was volume of the ipsilateral limb measured opto-electronically using a perometer and expressed as a percentage of the contralateral limb volume. RESULTS: At 12 months post-randomisation, there was no significant difference between treatment and control groups in terms of arm volume. Absolute change in arm volume at 12 months was 2.5% (95% CI -0.40 to 5.3) in the treatment group compared to 1.2% (95% CI -2.8 to 5.1) in the placebo group. The difference in mean volume change between randomisation groups at 12 months was not statistically significant (P = 0.6), -1.3% (95% CI -6.1 to 3.5), nor was there a significant difference in response at 6 months (P = 0.7), where mean change in arm volume from baseline in the treatment and placebo groups was 2.3% (95% CI -7.9 to 3.4) and -1.1% (95% CI -3.9 to 1.7), respectively. There were no significant differences between randomised groups in terms of secondary endpoints, including tissue induration (fibrosis) in the irradiated breast or chest wall, pectoral fold or supraclavicular fossa, change in photographic breast/chest wall appearance or patient self-assessment of function and Quality of Life at either 6 or 12 months. CONCLUSIONS: The study fails to demonstrate efficacy of dl-alpha tocopheryl acetate plus pentoxifylline in patients with arm lymphoedema following axillary surgery and lymphatic radiotherapy, nor does it suggest any benefits of these drugs in radiation-induced induration (fibrosis) in the breast, chest wall, pectoral fold, axilla or supraclavicular fossa. PMID- 15542160 TI - Is extended volume external beam radiation therapy covering the anastomotic site beneficial in post-esophagectomy high risk patients? AB - BACKGROUND AND PURPOSE: To assess the impact of extended volume radiation therapy (RT) with anastomotic coverage on local control in high risk post-operative esophageal cancer patients. PATIENTS AND METHODS: This is a retrospective study of high risk (T(3), T(4), nodes positive, with or without margin involvement) post-operative esophageal cancer patients treated at London Regional Cancer Centre from 1989 to 1999. After esophagectomy, all patients received adjuvant combined modality therapy consisting of four cycles of fluorouracil-based chemotherapy, and loco-regional RT with or without coverage of the anastomotic site. RT dose ranged from 45 to 60 Gy at 1.8-2.0 Gy/fraction with treatment fields tailored to the pathologic findings and location of the anastomosis. CT planning was used in all patients to design spinal cord sparing beam arrangements. First relapse rate (first incidence of an event), disease specific survival and overall survival were calculated by Chi-Square, Log-Rank, and Kaplan Meier (K-M) methods. RESULTS: During the study period, 72 patients had underwent esophagectomy and were considered for adjuvant chemoradiation therapy. Three patients were excluded due to disease progression prior to therapy. The 69 remaining patients formed the study cohort for the present analysis. The median age of the study group was 60 years (range 35-82 years). Pathologic stage distribution (AJCC 1997 staging) was T(2,3) N(1) in 94% patients, 65% of the cases were adenocarcinoma and had undergone transhiatal esophagectomy (86%) with positive/close margins in 34 (49%) patients. Median follow-up was 30.5 months (range 3.4-116.3 months). Two- and 5-year actuarial overall survivals rates were 50 and 31%, respectively. First relapse rate after adjuvant therapy was 63.7% (n = 44) and median time to relapse was 27.2 months. Anastomosis recurrence rates were 29% with small volume and 0% with extended volume RT (P = 0.041). Local and regional relapse occurred in 74.2% of patients treated with small volume RT compared to 15.4% in patients treated with extended volume RT (P < 0.001). After adjusting for resection margin status, the local control benefit of extended volume RT remained significant (P = 0.003). Treatment interruptions and late gastrointestinal toxicity were not significantly increased with the use of extended volume RT. CONCLUSIONS: A significant decrease in local and regional relapse without added late toxicity was achieved with the use of extended volume RT encompassing the anastomotic site post-operatively in high risk esophageal cancer patients. PMID- 15542161 TI - The role of endoscopic ultrasound (EUS) in planning radiotherapy target volumes for oesophageal cancer. AB - Computed tomography (CT) scanning is the main imaging modality utilised for planning radical oesophageal radiotherapy. Endoscopic ultrasound allows accurate localisation and local staging of oesophageal tumours. A method of incorporating this information into the CT planning process is described. Gross tumour volume position changes that occur when using this technique are presented. PMID- 15542162 TI - Does aqueous or sucralfate cream affect the severity of erythematous radiation skin reactions? A randomised controlled trial. AB - BACKGROUND AND PURPOSE: Evidence on which to base decisions about the management of radiation skin reactions is lacking. The purpose of this study was to investigate whether sucralfate or aqueous cream reduced acute skin toxicity during radiotherapy to the head and neck, breast or anorectal area (phase A), and to evaluate the effect of hydrogels and dry dressings on moist desquamation (phase B). This paper presents the results of phase A. PATIENTS AND METHODS: Three hundred and fifty seven patients were randomised to apply aqueous cream, sucralfate cream or no cream to the irradiated area from day one of radical radiotherapy treatment. All patients were instructed to wash using unperfumed soap. Acute skin toxicity was measured using a modified radiation therapy oncology group (RTOG) score, reflectance spectrophotometry, patient diary card and dermatology life quality index (DLQI). A cost minimisation approach was used to compare the costs of each skin care approach. RESULTS: No consistent differences were found in the severity of skin reactions or levels of discomfort suffered by patients in each of the randomised groups. Patients with a higher body mass index, who smoked, received concomitant chemotherapy, boost or bolus during treatment were more likely to develop skin reactions. CONCLUSIONS: There is no evidence to support the prophylactic application of either of the creams tested for the prevention of radiation skin reactions. Our results show that it is possible to predict which patients are at greatest risk of skin reactions. We suggest that known risk factors should be incorporated into future study protocols. PMID- 15542163 TI - Radiotherapy for nasopharyngeal carcinoma--transition from two-dimensional to three-dimensional methods. AB - This review describes the clinical background that underlies the transition from two-dimensional to three-dimensional (3D) planning techniques in the treatment of nasopharyngeal cancer (NPC). A systematic search of the Medline was performed using 'nasopharyngeal carcinoma', 'radiotherapy', '3-dimensional conformal radiotherapy', 'stereotactic radiosurgery/radiotherapy' and 'intensity-modulated radiotherapy' as keywords. Citing evidence from the published literature and their own institutional experience, the authors critically examined the positive impact of 3D methods--with emphasis on intensity-modulated radiotherapy (IMRT)- on target coverage and geometric accuracy, sparing of normal organs, and dosimetric homogeneity. Potential problems related to the widespread practice of IMRT such as quality assurance, utilization of medical resources and the risk of developing radiation-induced secondary cancers were highlighted. Application of IMRT within the context of altered fractionation, dose escalation and concurrent chemotherapy were discussed. The article concluded with a suggested treatment approach and research direction for different stages of NPC. PMID- 15542164 TI - Hyperfractionated accelerated radiochemotherapy (HFA-RCT) with mitomycin C for advanced head and neck cancer. AB - To investigate efficacy and feasibility of hyperfractionated accelerated radiotherapy combined with mitomycin C, patients with locally advanced unresectable squamous cell carcinomas of the head and neck region were administered 64-66 Gy in four weeks and mitomycin C (20 mg/m(2)) on day five. Twenty-one consecutive patients were included between November 1997 and June 1999 (median age: 57 years). All tumours were stage T3-4 and 18/21 were N2-3. Eighteen patients experienced grade 3 and three patients grade 2 mucosal toxicity. With median follow up for surviving patients of 42 months, loco-regional control was 55% at three years, overall survival was 33% at three years. This treatment is at the edge of local tolerability, but there is a good curative chance even for very advanced localised tumours, provided a complete remission is induced at primary treatment. PMID- 15542166 TI - Correlation between the treated volume, the GTV and the CTV at the time of brachytherapy and the histopathologic findings in 33 patients with operable cervix carcinoma. AB - BACKGROUND AND PURPOSE: This study correlates the treated volume, the GTV and the CTV at the time of intracavitary brachytherapy (BT) with the histopathological findings obtained by surgery (S) in 33 patients (pts) with cervix carcinoma. PATIENTS AND METHODS: Sixteen pts (group I), FIGO stage IB1 (1), IB2 (4), IIB (10), IIIB (1), received external beam radiotherapy (EBT) with a total dose of 45 Gy in 5 weeks and concomitant CISPLATIN 40 mg/m(2) weekly, followed by BT up to a total dose of 15 Gy. S was performed 6-8 weeks thereafter. Seventeen pts (group II), FIGO IA2 (1), IB1 (14), IIB (2), were treated by BT alone with a total dose of 60 Gy and S after 6-8 weeks. All pts had a MRI examination after BT with a moulded applicator in situ for exact delineation of GTV, CTV and critical organs and a 3D dosimetry directly from MRI data. RESULTS: In group I (EBT + BT + S), the histopathological findings showed complete tumour sterilization (CR) in 56% of pts. Residual disease (RD) was found in 43%. Dosimetric data showed in pts with CR a larger mean treated volume (213 vs. 166 cm(3)) and a better mean coverage of the GTV and the CTV by the reference isodose (99 and 91%) as in pts with RD (85 and 77%). In group II (BT + S), CR was found in 52%, RD in 41%. Dosimetric data showed a larger mean treated volume (154 vs. 109 cm(3)) for pts with RD and a mean coverage of the GTV and the CTV by the reference isodose of 97 and 84% vs. 89 and 80% for pts with CR. CONCLUSIONS: An incomplete coverage of the GTV and/or the CTV by the reference isodose is an important risk factor for RD at the time of surgery. Furthermore, for pts who received BT alone, tumour size seemed to be a limiting factor for an accurate coverage of the CTV by the reference isodose. PMID- 15542165 TI - Reduced recurrence of late hemorrhagic radiation cystitis by WF10 therapy in cervical cancer patients: a multicenter, randomized, two-arm, open-label trial. AB - BACKGROUND AND PURPOSE: To evaluate the efficacy and the safety of WF10 as adjunct to standard treatment in the management of late hemorrhagic radiation cystitis compared to standard treatment alone. PATIENTS AND METHODS: Cervical cancer patients with Grade 2 or 3 late hemorrhagic radiation cystitis, were randomized and treated with WF10 0.5 ml/kg body weight, diluted in physiological saline or 5% dextrose water 250 ml, intravenous infusions over 2 h on 5 consecutive days, every 3 weeks for 2 cycles plus standard treatment (WF10 group) or standard treatment alone (control group). Fifty patients in each group were evaluated by questioning; urinalysis and cystoscopy during a 1 year follow up. RESULTS: At week 7, 37 patients (74%) in the WF10 group and 32 patients (64%) in the control group showed complete resolution in objective hematuria (P = 0.28). Significantly lower use of antibiotics (P = 0.002) and antispasmodics (P < 0.001) was found in the WF10 group. Among the responders, 24 patients (77%) in the control group experienced recurrent objective hematuria, whereas in the WF10 group only 17 patients (47%) experienced a recurrence (P = 0.01). Recurrence of objective hematuria occurred significantly faster in the control group as evidenced by Kaplan-Meier and log-rank statistics (P = 0.004), suggesting a long term effect of WF10. Cystoscopy, at the end of the treatment period and after the one year follow up showed overall improvement without significant difference between two groups. No severe toxicity was monitored. CONCLUSIONS: WF10 therapy is a safe, non-invasive and convenient method in the management of late hemorrhagic radiation cystitis. WF10 therapy, as adjunct to standard treatment, has significantly reduced recurrence of objective hematuria, compared to standard treatment alone, during a one year follow up. PMID- 15542167 TI - High dose rate afterloading intraluminal brachytherapy for advanced inoperable rectal carcinoma. AB - BACKGROUND AND PURPOSE: High dose rate intraluminal brachytherapy for tumours of the rectal and anal canal which were inoperable either because of the age and frailty of the patient or because of advanced disease has been evaluated. PATIENTS AND METHODS: In a retrospective review of 50 consecutive patients the two main indications for brachytherapy were as part of a radical radiation programme in those unfit for major surgery (26 patients) or as palliation for advanced or metastatic disease (22 patients). Radical treatment was either sole treatment delivering 6 Gy fraction 2 to 3 times weekly up to 36 Gy or as a boost of 12 Gy after 45 Gy in 25 fractions external beam chemoradiation. Palliative treatments were given predominantly as a single dose of 10 Gy. RESULTS: This was predominantly a group of frail elderly patients with a median age of 82 years (range 35-91). Local tumour response was seen in 21/25 assessable patients with 14 complete responses. Median survival for the entire population was 6 months (range 1-54 months); in patients treated with 'radical' intent this was 25 months (range 1.5-54) and in the palliative group 7.2 months (range 1-37). The most common presenting symptom was bleeding per rectum for which a 64% response rate was obtained with 57% complete responses. Mucous discharge responded in 64% with 28% complete responses. The median duration of response was 7 months. CONCLUSION: Intraluminal HDR brachytherapy is an effective local treatment for patients otherwise unfit for radical surgery both as a component of radical treatment, or as a simple single palliative procedure. PMID- 15542168 TI - Sensitivity of treatment plan optimisation for prostate cancer using the equivalent uniform dose (EUD) with respect to the rectal wall volume parameter. AB - BACKGROUND AND PURPOSE: To analyse the sensitivity of plan optimisation of prostate cancer treatments with respect to changes in the volume parameter (n), when the EUD is used to control the dose in the rectal wall. PATIENTS AND METHODS: A series of plans was defined, by varying n over a range between 0.08 and 1, and testing different cost functions and beam arrangements. In all cases, the aim was to minimise the EUD in the rectal wall, while ensuring specific dose coverage of the PTV, and limiting the dose in the other OARs. The results were evaluated in terms of 3-D dose distribution and with respect to the current clinical knowledge about late rectal toxicity after irradiation. RESULTS: Different values of n lead to very similar dose distributions over the PTV (differences in mean dose < 1 Gy, differences in dose given to 99% of the volume < 1%). For the rectal wall, the following observations were made: (a) all cumulative DVH curves crossed each other around 60 Gy; (b) the rectal wall volume receiving doses between 30 and 45 Gy could change by 45 and 30%, respectively, depending on the value of n; (c) for doses higher than 70Gy the differences were typically within 5%. Different values of n also affected the position of isodose surfaces. The distance between the 70 and the 30 Gy isodose curves changed in the AP direction by a factor of 3 when n decreased from 1 to 0.08. High values of n were associated with less dose conformity and a larger volume (at least 20%) of normal tissues receiving 50 Gy or more. All DVHs for the rectal wall were below published dose toxicity thresholds except when the prescribed dose was escalated up to 86 Gy. CONCLUSIONS: In most cases, the solutions associated with n values up to 0.25 produced similar dose distribution in the rectal wall for doses above 45 Gy, complying with the dose-toxicity thresholds we analysed. The choice of a specific value of n in the optimisation requires an analysis of its effects on the dose distribution for the rectal wall, but also on other aspects, such as the value of the dose to the non-involved normal tissues. PMID- 15542169 TI - High dose three-dimensional conformal boost (3DCB) using an orthogonal diagnostic X-ray set-up for patients with gynecological malignancy: a new application of real-time tumor-tracking system. AB - The feasibility and accuracy of high dose three-dimensional conformal boost (3DCB) using three internal fiducial markers and a two-orthogonal X-ray set-up of the real-time tumor-tracking system on patients with gynecological malignacy were investigated in 10 patients. The standard deviation of the distribution of systematic deviations (Sigma) was reduced from 3.8, 4.6, and 4.9 mm in the manual set-up to 2.3, 2.3 and 2.7 mm in the set-up using the internal markers. The average standard deviation of the distribution of random deviations (sigma) was reduced from 3.7, 5.0, and 4.5 mm in the manual set-up to 3.3, 3.0, and 4.2 mm in the marker set-up. The appropriate PTV margin was estimated to be 10.2, 12.8, and 12.9 mm in the manual set-up and 6.9, 6.7, and 8.3 mm in the gold marker set-up, respectively, using the formula 2Sigma + 0.7sigma. Set-up of the patients with three markers and two fluoroscopy is useful to reduce PTV margin and perform 3DCB. PMID- 15542170 TI - Intensity modulated radiation therapy with electrons using algorithm based energy/range selection methods. AB - BACKGROUND AND PURPOSE: In recent years photon intensity modulated radiation therapy (IMRT) has gained attention due to its ability to improve conformity of dose distributions. A potential advantage of electron-IMRT is that the dose fall off in the depth dose curve makes it possible to modulate the dose distribution in the direction of the beam by selecting different electron energies. This paper examines the use of a computer based energy selection in combination with the IMRT technique to optimise the electron dose distribution. MATERIALS AND METHODS: One centimetre square electron beamlets ranging from 2.5 to 50 MeV were pre calculated in water using Monte Carlo methods. A modified IMRT optimisation tool was then used to find an optimum mix of electron energies and intensities. The main principles used are illustrated in some simple geometries and tested on two clinical cases of post-operated ca. mam. RESULTS: It is clearly illustrated that the energy optimisation procedure lowers the dose to lung and heart and makes the dose in the target more homogeneous. Increasing the energy at steep gradients compensates for lack of target coverage at beam edges and steep gradients. Comparison with a clinically acceptable four segment plan indicates the advantage of the used electron IMRT technique. CONCLUSIONS: Using an intensity optimised mix of computer selected electron energies has the potential to improve electron treatments for mastectomy patients with good target coverage and reduced dose to normal tissue such as lung and heart. PMID- 15542171 TI - Adjuvant high dose rate brachytherapy (Ir-192) in the management of keloids which have recurred after surgical excision and external radiation. AB - We describe our experience with adjuvant high dose rate brachytherapy (Ir-192) (HDRB) in patients, who failed surgery and post-operative external radiation therapy. The salvage treatment consisted of excision of the keloid and wound closure followed by HDRB (15 Gy in three fractions given on three consecutive business days beginning the day of surgery). At the time of last follow up, 88% (15/17) of the keloids were without any evidence of recurrence. PMID- 15542172 TI - Audit of effectiveness of routine follow-up clinics after radiotherapy for cancer: a report of the REACT working group of ESTRO. AB - BACKGROUND AND PURPOSE: The European Society for Therapeutic Radiology and Oncology was funded by the EU for a project on recording providing education, and ameliorating the consequences of treatment (REACT). An European audit was carried out as part of which to assess the usefulness of current follow-up practices. PATIENTS AND METHODS: Over a 4-month period in 15 cancer centres in 10 countries, patients attending for routine follow-up completed a questionnaire covering their expectations of and satisfaction with the visit. This was matched with a questionnaire completed by the physician about the content and usefulness of the consultation. The feasibility of a short toxicity scale developed by Dische and Saunders was also investigated. RESULTS: In total, 2303 matched questionnaires were analysed. Forty percent of the patients had symptoms or medical problems related to their disease. In 18% there was a positive finding on clinical examination. In 28% investigations were undertaken part of departmental routine practice. Ten percent of the investigations showed an abnormal result. Ninety nine percent of physicians and 85% of the patients expressed satisfaction. Using the short toxicity scale rates of recording toxicity could be increased from 28 to 93%. CONCLUSIONS: There is wide variation in follow-up practices among European centres. There was a low incidence of positive findings clinically or with routine investigations. A simple scale for recording morbidity has proved easy to use by departments, which have not routinely used one of the standard measures. Further work will attempt to produce an European guideline for effective routine follow-up after radiotherapy. PMID- 15542173 TI - Re: new radiotherapy technologies for meningiomas: 3D conformal radiotherapy? Radiosurgery? Sterotactic radiotherapy? Intensity modulated radiotherapy? Proton beam radiotherapy? Spot scanning proton radiation therapy? Or nothing at all? [Radiother Oncol 2004;71(3):247-249]. PMID- 15542174 TI - Re: Vikram, B., The PSA conundrum. [Radiother Oncol, 2004. 71(1): p. 1-2]. PMID- 15542176 TI - Henoch-Schonlein purpura with antiphospholipid antibodies after influenza vaccination: how fearful is it in children? PMID- 15542177 TI - Post-marketing surveillance of immediate allergic reactions: polygeline-based versus polygeline-free pediatric TBE vaccine. AB - Scattered cases of immediate allergic reactions occurred in the nineties after widespread use of the original (polygeline-based) pediatric tick-borne encephalitis (TBE) vaccine and were reported to Pharmacovigilance, Chiron Vaccines. Although, still indicating a very rare frequency of about two cases per 100,000 doses sold, the benefit/risk assessment resulted in its withdrawal from the market in early 1998. An intensive evaluation revealed that polygeline used as a vaccine stabilizer was the most probable cause of the reported allergic reactions. Consequently, an improved pediatric TBE vaccine, free of polygeline and other protein-derived vaccine stabilizers, was developed. A post-marketing surveillance analysis covering the first two vaccination seasons after the introduction of this new pediatric TBE vaccine in early 2002 reveals a very low reporting rate of immediate allergic reactions post immunization (within the range as noted for other widely used vaccines for childhood immunization), i.e., 0.08-0.24 cases per 100,000 doses sold depending on case definition and medical assessment. In conclusion, this analysis provides post-marketing surveillance evidence that the change in the vaccine formulation, with regards to the potential risk of immediate allergic reactions, has led to an intended improvement in the vaccine's safety profile. PMID- 15542178 TI - Targeting of Porphyromonas gingivalis with a bispecific antibody directed to FcalphaRI (CD89) improves in vitro clearance by gingival crevicular neutrophils. AB - Phagocytosis and killing of pathogens by polymorphonuclear neutrophils (PMN) from gingival crevicular fluid (GCF) is diminished in chronic periodontitis patients. As an approach to improve targeting of PMN toward a periodontopathogen, Porphyromonas gingivalis, the efficacy of a bispecific antibody (BsAb) directed against both recombinant 130 kDa hemagglutinin domain (r130k-HMGD) of P. gingivalis, and PMN Fc receptor (FcR) was evaluated. GCF PMN exhibited higher IgA FcR (FcalphaRI) levels, and lower IgG FcR (FcgammaRIIa and FcgammaRIIIb) levels than PB PMN. Functional studies revealed that GCF PMN exhibited a higher capacity to phagocytose and kill P. gingivalis opsonized with a BsAb targeting P. gingivalis r130k-HMGD to FcalphaRI as compared to an anti-r130k-HMGD antibody. However, phagocytosis and killing activity of PB PMN that were incubated with the two antibodies proved comparable. These data support targeting of pathogens toward FcalphaRI as an option to improve antibacterial immunity in chronic periodontitis patients. PMID- 15542179 TI - Oral vaccination of pigs with an invasive gyrA-cpxA-rpoB Salmonella Typhimurium mutant. AB - The potency to protect pigs against colonization and against clinical salmonellosis was evaluated after oral immunization with a live gyrA-cpxA-rpoB Salmonella (S.) Typhimurium mutant (S. Tm. Nal2/Rif9/Rtt). Twenty 4-week-old male hybrid piglets were immunized orally, a control group received a placebo. Three weeks postimmunization, all pigs were challenged orally with a highly virulent S. Typhimurium DT104 strain. Clinical investigation revealed that immunization prevented the vaccinated pigs from clinical symptoms of salmonellosis. While all placebo-treated animals showed a 2-4-day episode of moderate to severe clinical symptoms, 90% of immunized pigs did not show any clinical signs at all. The bacteriological results showed a marked beneficial effect of the oral immunization. Vaccinated pigs showed a significantly decreased rate of colonization of the inner organs (42.5% versus 87.5%) when compared to the placebo-treated animals. Furthermore, in comparison to the non-immunized pigs, the vaccines developed a higher specific immunoglobulin (Ig)A antibody activity, but a significant lower IgM antibody activity in serum. The findings underline the ability of an attenuated oral live S. Typhimurium mutant to prevent clinical symptoms of salmonellosis in pigs and to significantly reduce the colonization of tissues and inner organs, as well as the shedding of S. Typhimurium. PMID- 15542180 TI - Bordetella pertussis adenylate cyclase delivers chemically coupled CD8+ T-cell epitopes to dendritic cells and elicits CTL in vivo. AB - The adenylate cyclase (CyaA) produced by Bordetella pertussis is able to deliver CD8+ and CD4+ T-cell epitopes genetically grafted within the catalytic domain of the molecule into antigen presenting cells in vivo. We develop now a new approach in which peptides containing CD8+ epitopes are chemically linked to CyaA. We show that CTL responses were induced in mice immunized with CyaA bearing these CD8+ epitopes. Moreover, we demonstrate that the OVA257-264 CD8+ epitope chemically grafted to CyaA is presented to CD8+ T cells by a mechanism requiring (1) proteasome processing, (2) TAP and (3) neosynthesis of MHC class I molecules. Thus, this novel strategy represents a very versatile system as a single CyaA carrier protein could be easily and rapidly coupled to any desired synthetic peptide. PMID- 15542181 TI - Co-administration of CpG oligonucleotides enhances the late affinity maturation process of human anti-hepatitis B vaccine response. AB - We assessed the avidity maturation process elicited by human immunization with alum-adsorbed HBsAg alone or with a novel adjuvant containing CpG motifs (CpG 7909). Mean avidity indexes and distribution of low- and high-avidity anti-HBs indicated that avidity maturation essentially takes place late after priming. CpG 7909 markedly enhanced this affinity maturation process, increasing the pool of high-avidity antibodies. The influence of CpG 7909 was antigen-specific, isotype specific and distinct from the influence on anti-HBs production, as avidity did not correlate with anti-HBs IgG titers. This is the first demonstration that a novel human adjuvant may induce antibodies with higher antigen-binding affinity. PMID- 15542182 TI - Risk factors for acellular and whole-cell pertussis vaccine failure in Senegalese children. AB - Although use of acellular pertussis vaccine was associated with a higher rate of vaccine failure than that of whole-cell vaccine in the Senegal Pertussis Trial conducted in 1990-1994 on 4189 children, risk factors for vaccine failure regarding exposure and susceptibility to pertussis have not been studied so far. Pertussis occurred in 346 vaccinated children. Three factors were found to be associated with vaccine failure, independently of the vaccine type, namely the degree of exposure, birth rank, and time since weaning. In the whole-cell vaccine group, the risk of failure increased with birth rank [RR = 2.95 (1.51-5.75)] and was higher in non stunted children [RR = 1.43 (1.05-1.94)]. In the acellular vaccine group, the risk of failure increased with age at exposure to B. pertussis [RR = 2.24 (1.21-4.12) after 18 months of age] and the degree of exposure [RR = 2.14 (1.17-3.93) when the child shared the hut of an index case]. These results highlight the influence of environmental factors on the success of pertussis vaccination. However, they do not explain the shorter duration of protection provided by the acellular vaccine compared to the whole-cell vaccine which persist after controlling and thus might be related to the nature of the vaccine. PMID- 15542183 TI - Plasmid encoding papillomavirus Type 16 (HPV16) DNA constructed with codon optimization improved the immunogenicity against HPV infection. AB - Human papillomavirus Type 16 (HPV16) infections can cause neoplasia, which is thought to be closely associated with the development of cervical cancers. In the study, we attempted to construct a DNA plasmid encoding a HPV16 capsid protein (L1) and a HPV16 oncoprotein (E7), which was capable of preventing HPV16 infection and eliminating HPV16-infected cells. A plasmid, L1E7hpSCA1, encoding the L1 and E7 genes with the codon usage optimized for mammalian cell expression, was constructed. Mutations were introduced into the E7 gene sequence for reducing its oncogenicity. C57BL/6 mice were intramuscularly immunized at tibialis anterior (TA) muscles with the newly constructed L1E7hpSCA1 plasmid. The immune responses induced by the L1E7hpSCA1 plasmid (with codon optimization) and a control L1E7pSCA1 plasmid (without codon optimization) were compared. It is shown that the L1E7hpSCA1 was able to induce much stronger immune responses than the L1E7pSCA1. Sera obtained from immunized animals were found to contain anti-HPV16 antibodies as detected by ELISA and hemagglutination inhibition (HAI) assays. Cytotoxicity and interferon-gamma assays showed that spleenocytes from immunized animals were able to recognize and lyze E7 expressing tumor TC-1 cells. Moreover, the growth of E7 expressing tumor mass was inhibited in vaccinated mice. In vivo tumor protection test indicated that tumor formation was prevented in the experimental animals (67%) after vaccination with L1E7hpSCA1, while for the control group injected with L1E7pSCA1 only and the animal group injected with pSCA1 only, tumor formation was observed in all experimental animals. Our results suggest that the L1E7h gene (with codon optimization) is more effective against HPV16 than the L1E7 gene (without codon optimization). The L1E7hpSCA1 plasmid was able to provide protection against E7 expressing tumor, and it might have the potential to be a vaccine candidate for HPV prevention. PMID- 15542184 TI - Is influenza vaccination cost effective for healthy people between ages 65 and 74 years? A randomised controlled trial. AB - The aim of this study was to determine the cost effectiveness of influenza vaccination for healthy people aged 65-74 years living in the UK. People without risk factors for influenza (chronic heart, lung or renal disease, diabetic, immunosuppressed or those living in an institution) were identified from 20 general practitioner (GP) practices in Liverpool in September 1999. 729/5875 (12.4%) eligible individuals were recruited and randomised to receive either influenza vaccine or placebo (ratio 3:1), with all participants receiving 23 valent-pneumococcal polysaccharide vaccine unless already administered. The primary analysis was the frequency of influenza as recorded by a GP diagnosis of pneumonia or influenza like illness. In 2000, the UK vaccination policy was changed with influenza vaccine becoming available for all people aged 65 years and over irrespective of risk. As a consequence of this policy change, the study had to be fundamentally restructured and only results obtained over a one rather than the originally planned two-year randomised controlled trial framework were used. Results from 1999/2000 demonstrated no significant difference between groups for the primary outcome (relative risk 0.8, 95% CI 0.16-4.1). In addition, there were no deaths or hospitalisations for influenza associated respiratory illness in either group. The subsequent analysis, using both national and local sources of evidence, estimated the following cost effectiveness indicators: (1) incremental NHS cost per GP consultation avoided = 2000 pound sterling; (2) incremental NHS cost per hospital admission avoided = 61,000 pound sterling; (3) incremental NHS cost per death avoided = 1,900,000 pound sterling and (4) incremental NHS cost per QALY gained = 304,000 pound sterling. The analysis suggested that influenza vaccination in this population would not be cost effective. PMID- 15542185 TI - Bovine respiratory syncytial virus ISCOMs--protection in the presence of maternal antibodies. AB - The protection induced by immunostimulating complexes (ISCOMs) against bovine respiratory syncytial virus (BRSV) was evaluated and compared to that of a commercial inactivated vaccine (CV) in calves with BRSV-specific maternal antibodies. Following experimental challenge, controls (n = 4) and animals immunized with CV (n = 5) developed moderate to severe respiratory disease, whereas calves immunized with ISCOMs (n = 5) remained clinically healthy. BRSV was re-isolated from the nasopharynx of all controls and from all calves immunized with CV, but from none of the calves immunized with ISCOMs. BRSV-RNA was detected by real-time PCR from a single animal in this group. Significantly higher BRSV-specific nasal IgG, serum IgG1 and IgG2 titers were detected before and after challenge in animals immunized with ISCOMs versus CV. In conclusion, the ISCOMs overcame the suppressive effect of maternal antibodies in calves and induced strong clinical and virological protection against a BRSV challenge. PMID- 15542186 TI - Safety and immunogenicity of a booster dose of Staphylococcus aureus types 5 and 8 capsular polysaccharide conjugate vaccine (StaphVAX) in hemodialysis patients. AB - StaphVAX, an unadjuvanted, bivalent vaccine composed of Staphylococcus aureus (S. aureus) capsular polysaccharides (CPS) types 5 and 8 bound to the mutant non toxic recombinant Pseudomonas aeruginosa exotoxin A (rEPA) conferred approximately 60% protection for 10 months against bacteremia caused by this pathogen in hemodialysis patients. A protective level of 80 microg/ml was estimated based upon geometric mean (GM) antibody levels at the end of the efficacy period. To extend the duration of protection conferred by StaphVAX in hemodialysis patients, recipients of the vaccine were reinjected in a randomized double-blinded, placebo-controlled study. Vaccinees received StaphVAX and a saline placebo injection 14 days apart according to the randomization schedule. The booster dose of StaphVAX was administered an average of 958 days (753-1167 days) after the first injection. There were no serious adverse reactions. Antibody levels at day 14, 28, 92, and 182 post-injection were measured by ELISA. Maximal levels of IgG anti-CPS were observed at the 28-day interval. For type 5, GM antibody levels increased from 73 microg/ml at day 0 to 162 microg/ml (P < 0.001) and for type 8 from 59 microg/ml to 133 microg/ml (P < 0.001). Anti-CPS antibody levels of approximately 80 microg/ml to type 5 and type 8 were achieved in 72.4 and 74.3% of vaccinees, respectively. There was excellent correlation between the level of anti-CPS and opsonic titer (r = 0.93). Moreover, the decline of anti-CPS antibody levels at six months was significantly less rapid than that observed from the first immunization (P < 0.001). We conclude that a booster immunization to maintain protective levels of specific antibodies for an extended period of time is feasible for patients at continuous risk for S. aureus bacteremia. PMID- 15542187 TI - Vaccination of persons allergic to latex: a review of safety data in the Vaccine Adverse Event Reporting System (VAERS). AB - Vaccine products currently licensed in the US and other countries are marketed in vials and syringes that may contain natural latex allergens. Little scientific information exists regarding the safety of vaccination of latex-allergic individuals. A review of data within the Vaccine Adverse Event Reporting System (VAERS), a large registry of reported possible vaccine adverse reactions was conducted. A search of the database, which contains >160,000 vaccine adverse event reports, revealed only 28 cases of possible immediate-type hypersensitivity reactions in vaccine recipients with a history of allergy to latex. Given the large number of immunizations administered every year in the US, the reported risk of allergic reactions possibly due to latex contamination of vaccines appears to be very small. PMID- 15542188 TI - Spontaneously reported adverse reactions after diphtheria-tetanus revaccination at 4-6 years of age--a comparison of two vaccines with different amounts of diphtheria toxoid. AB - On 1 January 1996, diphtheria-tetanus revaccination at the age of 5 years was implemented in the Danish Childhood Vaccination Programme. Initially, a combined DT vaccine containing 25 Lf diphtheria toxoid was used. Due to a high frequency of spontaneously reported adverse reactions, however, concerns were raised that the diphtheria dose was too high, and it was reduced to 6.25 Lf. This survey presents the rates of spontaneously reported adverse reactions following diphtheria-tetanus revaccinations of 4-6-year-old from 1996 to 2002. The change to the lower dose of diphtheria toxoid resulted in a remarkable reduction in the yearly rates of injection site reactions: in 1996, the rate was 180 injection site reactions per 100,000 vaccinations; from 1998 to 2002, this changed to between 12 and 24 reactions per 100,000 vaccinations. Furthermore, the rates of systemic reactions such as fever were reduced. PMID- 15542189 TI - Cationic microparticles are a potent delivery system for a HCV DNA vaccine. AB - We initially evaluated in mice the ability of naked DNA encoding intracellular forms of the E1E2 envelope proteins from HCV to induce antibody responses and compared the responses induced with the same plasmid adsorbed onto cationic poly (lactide co-glycolide) (PLG) microparticles. Although naked DNA was only able to induce detectable responses at the 100 microg dose level, making this approach impractical for evaluation in larger animals, PLG/DNA induced detectable responses at 10 microg. In addition, the PLG/DNA microparticles induced significantly enhanced responses to naked DNA when compared at the same dose level. Remarkably, PLG/DNA induced comparable responses to recombinant E1E2 protein adjuvanted with the emulsion MF59. Furthermore, PLG/DNA effectively primed for a booster response with protein immunization, while naked DNA did not. Therefore, PLG/DNA was selected for further evaluation in a non-human primate model. In a study in rhesus macaques, PLG/DNA induced seroconversion in 3/3 animals following three immunizations. Although the antibody responses appeared lower than those induced with recombinant protein adjuvanted with MF59, following a fourth dose, PLG/DNA and protein induced comparable responses. However, a single booster dose of recombinant protein administered to the animals previously immunized with PLG/DNA induced much higher responses. In addition, one of three animals immunized with PLG/DNA showed a cytotoxic T lymphocyte response in peripheral blood lymphocytes. In conclusion, cationic PLG microparticles with adsorbed HCV DNA generates potent immune responses. PMID- 15542190 TI - Epidermal powder immunization against influenza. AB - Epidermal powder immunization (EPI) can efficiently deliver powdered protein vaccines to the epidermis. A phase I clinical trial was conducted to evaluate powdered trivalent influenza vaccine delivered using the PowderJect ND5.2 delivery system. Subjects received either Fluvirin IM injection (15 microg of each influenza strain), a single EPI vaccination (15 microg of each influenza strain) or two adjacent EPI (total of 30 microg of each influenza strain). Systemic reactogenicity was similar between control and EPI vaccines. Site reactions following EPI were primarily mild and self-limiting. Seroconversions, titer increases and geometric mean titers to all strains were equivalent or higher in EPI-immunized groups than in controls. Powdered influenza vaccine delivered by EPI is safe and elicits humoral immune responses in humans. PMID- 15542191 TI - Lack of induction of autoantibody responses following immunization with cytomegalovirus (CMV) glycoprotein B (gB) in healthy CMV-seronegative subjects. AB - Possible correlations have been proposed between autoimmune diseases, such as systemic lupus erythematosus (SLE), and infection with human cytomegalovirus (CMV). The recent observation that an adenovirus expressing the immunodominant envelope glycoprotein of CMV, glycoprotein B (gB), may be capable of inducing autoantibodies in certain mouse strains has prompted interest in exploring potential relationships between gB immunization and autoimmune disease. We examined whether a recombinant CMV gB vaccine, or a gB canarypox vectored vaccine (ALVAC-CMVgB), administered to a total of 76 CMV-seronegative subjects, was capable of inducing cross-reactive antibodies to Smith antigen (Sm), ribonucleoprotein complex (RNP), and the U1-70 kDa component of the RNP complex. Using immunofluorescence, EIA and immunoblot analyses, we failed to identify induction of autoantibodies following vaccination with gB, whether administered alone as a purified protein subunit with adjuvant, or in combination with expression in a vectored approach using a recombinant canarypox. These data reinforce the favorable safety profile of CMV gB vaccines. PMID- 15542192 TI - Structure and adsorption properties of commercial calcium phosphate adjuvant. AB - Calcium phosphate adjuvant is a commercially available vaccine adjuvant that potentiates the immune response to antigens. Although its name suggests that it is Ca3(PO4)2, X-ray diffraction, FTIR spectroscopy, thermal analysis and the Ca/P molar ratio identify commercial calcium phosphate adjuvant as non-stoichiometric hydroxyapatite, Ca10-x (HPO4)x (PO4)6-x (OH)2-x, where x varies from 0 to 2. The surface charge is pH-dependent (point of zero charge = 5.5). Consequently, commercial calcium phosphate adjuvant exhibits a negative surface charge at physiological pH and electrostatically adsorbs positively charged antigens. The presence of hydroxyls allows calcium phosphate adjuvant to adsorb phosphorylated antigens by ligand exchange with surface hydroxyls. PMID- 15542193 TI - Genetically programmed B lymphocytes are highly efficient in inducing anti-virus protective immunity mediated by central memory CD8 T cells. AB - The hallmarks of specific T cell immunity include proliferative expansion, acquisition of effector function and memory T cell formation. Here, we used priming with B lymphocytes transgenic for the dominant epitope (NP366-374) of the influenza virus nucleoprotein, to study the characteristics of the CD8 T cell memory response in C57Bl/6 mice and elucidate which subset of CD8 T cells memory mediates protection from disease. We found that (i) the size of the memory CTL response is independent of the priming dose and is similar to that induced by the live virus, (ii) priming with a low dose (3 x 10(2)cells/inoculum) of transgenic B lymphocytes confers a protective memory CTL response, and (iii) protection from disease is mediated by central memory (T(CM)) CD8 T cells. PMID- 15542194 TI - Efficacy of an Escherichia coli J-5 mutant strain bacterin in the protection of calves from endotoxin disease caused by subcutaneous challenge with endotoxins from Escherichia coli. AB - The purpose of this trial was to examine the potential of a new Escherichia (E) coli J-5 mutant strain bacterin to reduce the severity of clinical disease caused by subcutaneous challenge with endotoxins of Gram-negative bacteria in calves. Day-old to 3-day old calves (n = 40 per study phase) were randomly assigned to either of two treatment groups, i.e. a vaccinated or a placebo group. Calves in the vaccinated group received an inactivated bacterin containing a J-5 mutant strain of E. coli via subcutaneous route at 2-4 days of age and at 14 days thereafter. The placebo contained only adjuvant and saline in lieu of the antigen. Lipopolysaccharides (LPS) originating from E. coli were administered subcutaneously 3 weeks after the booster dose. The LPS challenge dosages were 1 and 8 microg/kg in study phases I and II, respectively. Various clinical, physiological, hematological, and serological parameters were measured at specific time intervals after challenge. The data were mostly analysed using peak changes from baseline recorded during the observation period. By the time of challenge the titers in vaccinated calves had increased significantly more than in the unvaccinated controls. Disease severity following subcutaneous challenge was dose dependent. In phase I, placebo calves were only mildly challenged whereas in phase II placebo calves showed a moderate challenge. After a mild challenge, there was little evidence of protection due to vaccination as only attitude was significantly improved in the vaccinates. In contrast, after a moderate challenge rectal temperature, hematocrit, blood glucose concentrations, and leukocyte changes were significantly better in the vaccinated group. In conclusion, the results of this study show that following a subcutaneous endotoxin challenge that induces a moderate clinical response, calves that were previously vaccinated with the E. coli J-5 bacterin were better protected than those in the placebo group. PMID- 15542195 TI - Human antibodies to recombinant protein constructs of Plasmodium falciparum Apical Membrane Antigen 1 (AMA1) and their associations with protection from malaria. AB - Serum antibodies from 1071 people in two Kenyan villages were assayed using eight different recombinant Apical Membrane Antigen 1 (AMA1) protein constructs to investigate their role in naturally acquired immunity. In both communities, antibodies against the full-length ectodomain (both FVO and 3D7 allele constructs) prior to a malaria transmission season were significantly associated with protection from malaria in the following 6 months, even after adjusting for age and antibody reactivity to whole parasite (schizont) extract. However, these protective associations of antibodies were only seen among subjects that were parasite slide positive at the time of pre-season serum sampling. Competition ELISAs with the FVO and 3D7 allele constructs showed that antibodies can recognise either conserved or allele-specific epitopes in AMA1. Results encourage the development of an AMA1 vaccine based on the full-length ectodomain, and indicate that the function of human antibodies to allele-specific and conserved epitopes in AMA1 should be studied further. PMID- 15542196 TI - Estimating the force of measles virus infection from hospitalised cases in Lusaka, Zambia. AB - Estimates of the force of infection (the rate at which susceptible individuals acquire infection) are essential for modelling the transmission dynamics of infectious diseases and can be a useful tool in evaluating mass vaccination strategies. Few estimates exist of the force of infection of measles virus in sub Saharan Africa. A mathematical model was applied to age-specific recorded hospital admissions between September 1996 and September 1999 to estimate the force of measles virus infection in Lusaka, Zambia. The average force of infection was estimated to be 20% per year (95% confidence intervals (CI) 16.5, 23.5) which was insensitive to varying assumptions about vaccine coverage. The force of infection varied from year to year (P < 0.001) reflecting the cyclic pattern of measles incidence. The estimated probability of a case being hospitalised decreased with age, consistent with less severe disease in older children. Estimates of the force of infection using routinely available data were consistent with those based upon serological surveys in other sub-Saharan African countries. PMID- 15542197 TI - Functional memory CD8+ T cells can be generated in vivo without evident T help. AB - Synthetic cytotoxic T cell (CTL) epitope peptides provide an effective and safe means of vaccination against cancers and viruses, as these peptides can induce specific CD8+ effector T cells in vivo. However, the effector CD8+ T cells induced by the minimal CTL epitope peptides do not last past about 3 weeks after the induction and no functional memory CD8+ T cells are generated. It is held that simultaneous induction of CD4+ T cells by incorporating peptides containing T-helper epitopes in the vaccine at the time of primary vaccination are necessary for the induction of long-lived functional memory CD8+ T cells. We now report that, surprisingly, incorporation of medium length (>20 AA) peptides devoid of detectable T-helper epitopes in a minimal CTL epitope-based vaccine can also induce long-lasting functional tumour antigen specific memory CD8+ T cells that are capable of promoting protection against tumour challenge. This observation may have implications for the formulation of therapeutic anti-cancer and anti virus peptide vaccines where a strong induction of CD4 T help would be undesirable. PMID- 15542198 TI - Sucrose fatty acid sulphate esters as novel vaccine adjuvants: effect of the chemical composition. AB - Adjuvant activity of novel, synthetic sucrose derivatives towards a recombinant glycoprotein was determined in large, non-rodent animal species. Compared to antigen alone, up to 3000-fold higher virus neutralizing antibody titres (VNTs) and 10-fold higher cellular responses against classical swine fever virus were observed in pigs after two immunizations with the sucrose derivatives combined with a squalane-in-water emulsion. The chemical composition of the derivative was crucial and sucrose esters containing one sulphate and seven dodecanoic (C12) or decanoic (C10) esters exerted the highest adjuvanticity. Derivatives without sulphate, with fewer fatty acid esters or with shorter or longer alkyl esters were less effective. Strong adjuvant activity of these formulations was the result of synergistic collaboration between the sucrose ester and the squalane emulsion, as factor of increase in VNT by the individual components was between 4 and 34. Enhanced humoral and cell-mediated immune responses lasted for at least 24 weeks. We concluded that combinations of hydrophobic, negatively-charged sucrose fatty acid sulphate esters plus submicron emulsions of squalane-in-water are strong adjuvants for humoral and cell-mediated immunity and that these formulations are promising adjuvants for future vaccines containing poor immunogens. PMID- 15542199 TI - The burden and cost of hospitalised varicella and zoster in Australian children. AB - BACKGROUND: Economic analyses of varicella-zoster virus (VZV) immunisation are sensitive to the costs of hospitalised cases, so there is a need to validate VZV hospitalisation data. AIMS: To assess the accuracy of hospital VZV coding data and to apply these parameters to a population-based sample to estimate incidence and costs. METHODS: A 3-year retrospective chart review from one hospital to document clinical features and validate coding data. A separate 9-year analysis of discharge data from two hospitals draining a defined region of suburban Melbourne, with adjustment for miscoding and estimates of direct hospital costs. RESULTS: After correction for miscoding, 224 patients were admitted to one hospital over 3 years, 79% with varicella and 21% with zoster. Miscoding resulted in a 15% underestimate of zoster cases and a 4% overestimate of varicella cases. Thirty-six percent of varicella admissions compared to 80% of zoster admissions were immunocompromised and/or had chronic disease. Compared to otherwise-healthy patients, immunocompromised patients were admitted earlier in their illness and had lower complication rates. Forty-two percent of immunocompromised/chronic disease patients with varicella had a known exposure, usually from a family member. The incidence of hospitalised varicella and zoster in under 15-year olds was 15.7 and 1.8 per 100,000 per year, respectively. This suggests that there are 615 varicella hospitalisations and 72 zoster hospitalisations in this age group each year in Australia, at a total direct cost of over 2.2 million AU dollars. CONCLUSION: These results highlight the considerable burden of hospitalised zoster and the importance of immunising non-immune contacts of immunocompromised individuals. They also support previous estimates of the incidence of hospitalised varicella in Australian children and adolescents, although direct medical costs may be higher than those previously estimated. PMID- 15542200 TI - Persistence of antibody response to pneumococcal capsular polysaccharides in vaccinated long term-care residents in Brazil. AB - To evaluate the immunogenicity of 23-valent pneumococcal polysaccharide vaccine in 52 nursing homes residents aged > or = 60 years, IgG antibodies to serotypes 1, 5, 6B, and 8 were measured by ELISA and compared before, and 1 and 12 months following vaccination. A significant immunological response for all serotypes was observed at 1 month after vaccination. The mean increase in antibody concentration was highly variable and ranged from 1.6 to 2.7. After 1 year, the mean concentrations remained significantly higher than prior to vaccination for serotypes 1, 6B, and 8, although there was a decrease in all mean IgG concentrations. Antibody levels were higher in men than in women, before and after immunisation. Post-vaccination values tended to be lower among subjects aged >75 years. Reduction in IgG concentrations by 33% 1 year after vaccination suggests that revaccination of institutionalised elderly people may be needed. PMID- 15542201 TI - CD80 and CD86, but not CD154, augment DNA vaccine-induced protection in experimental bovine tuberculosis. AB - DNA vaccination is known to elicit robust cellular and humoral responses to encoded antigen. The co-administration of costimulatory molecules CD80 (B7-1), CD86 (B7-2) and CD154 (CD40L) has been shown to enhance immune responses in several murine models. The role of specific costimulatory molecules in non-rodent species remains incompletely characterized. In these studies, we demonstrate that the co-administration of CD80 and CD86, but not CD154, to an existing candidate subunit DNA vaccine (ESAT-6) against bovine tuberculosis, enhances protection after aerosol challenge with virulent Mycobacterium bovis. Additionally, we have shown that vaccination with M. bovis BCG is protective against tuberculosis following aerosol challenge in cattle. Two independent trials were conducted in cattle to determine the adjuvant effect of encoded antigen + CD80/CD86 and directly compare the adjuvant activities of CD80/CD86 to those of CD154. Co administration of either CD80/CD86 or CD154 enhanced ESAT-6-specific IFN-gamma responses as compared to animals vaccinated with ESAT-6 DNA alone. However, following aerosol challenge, only animals vaccinated with CD80/CD86 possessed decreased pathology of the lungs and associated lymph nodes, as measured by gross examination, radiographic lesion morphometry and bacterial recovery. Collectively, these results demonstrate that the co-administration of costimulatory molecules with a protective antigen target enhances bovine immune responses to DNA vaccination, and that CD80/CD86 is superior to CD154 in augmenting DNA vaccine-induced protection in experimental bovine tuberculosis. PMID- 15542202 TI - A polyvalent DNA vaccine expressing an ESAT6-Ag85B fusion protein protects mice against a primary infection with Mycobacterium tuberculosis and boosts BCG induced protective immunity. AB - In this study, we evaluated the protective efficacy of a DNA vaccine (pE6/85) expressing an ESAT6-Ag85B fusion protein against a primary Mycobacterium tuberculosis infection in mice. In short-term studies, vaccination with pE6/85 protected as well as Mycobacterium bovis BCG immunization with similar lung pathology and bacterial burdens detected 28 days after a low dose aerogenic challenge (>1.0 log(10) reduction relative to naives). In a survival experiment, the protection induced by pE6/85 immunization was also not significantly different than that elicited by BCG vaccination with the mean-times-to-death (+/ standard error of the mean) being 102+/-20, 271+/-32 and 299+/-14 days for naive, pE6/85 and BCG-vaccinated mice, respectively. Furthermore, boosting with pE6/85 but not BCG or a DNA vaccine cocktail at 1 year after an initial BCG immunization (when BCG-induced protection was declining), augmented protection in the lung at 15 and 18 months to levels detected at 3 months post-BCG vaccination. PMID- 15542203 TI - Characterization of antibodies to PspA and PsaA in adults over 50 years of age with invasive pneumococcal disease. AB - We characterized antibody responses to two Streptococcus pneumoniae surface proteins, PspA and PsaA, in 14 adults over 50 years of age hospitalized with invasive pneumococcal disease (IPD), and in two groups of age-matched controls (18 patients with invasive disease due to other microorganisms and 35 patients hospitalized for non infectious conditions). All patients with IPD and all control subjects had detectable antibodies to both proteins on hospital admission. Three weeks later, the geometric mean concentrations of antibodies to PspA and PsaA in IPD patients were respectively 10 and 25 times higher than on admission. In contrast, acute and convalescent antibody levels were similar in control patients with invasive diseases due to other microorganisms. PMID- 15542204 TI - A novel dry powder influenza vaccine and intranasal delivery technology: induction of systemic and mucosal immune responses in rats. AB - Intranasal (i.n.) vaccination represents an attractive non-invasive alternative to needle-based injection and provides superior protection at mucosal surfaces. However, new formulations are needed to improve efficacy and reduce the refrigerated storage and distribution requirements associated with standard liquid vaccines. Here, we describe a powder formulation of whole inactivated influenza virus and a novel i.n. delivery platform. The powder-formulated vaccine elicited a significant serum antibody response in rats that was at least as strong as that provided by the liquid vaccine administered i.n. or via intramuscular (i.m.) injection. Significant nasal IgA responses were also observed solely after i.n. delivery. This study demonstrates for the first time the generation of potent nasal mucosal and systemic immune responses using an i.n. delivered influenza vaccine powder and suggests an alternative approach to vaccination against influenza and other infectious diseases. PMID- 15542205 TI - Dose response of CRM197 and tetanus toxoid-conjugated Haemophilus influenzae type b vaccines. AB - High vaccine cost has limited use of conjugate vaccines in the developing world where the disease burden is greatest. Fixed fractional doses of Haemophilus influenzae type b (Hib) vaccines have been shown to be immunogenic, but dose responses of these vaccines in humans are needed to determine the lowest immunogenic dose as an option for lowering vaccine cost. We randomized children to receive one of five doses (0.625, 1.25, 2.5, 5.0 and 10 microg) of either a diphtheria CRM197 or tetanus toxoid-conjugated Hib vaccine. The children received a primary three-dose series at 6, 10, and 14 weeks of age and a booster dose at 9 months. Anti-PRP IgG antibodies were measured at each vaccination, at 18 weeks, and at one week following the booster dose. Concentrations of > or =1.25 microg of HibCRM197 vaccine produced mean anti-PRP responses at 18 weeks of > or =5.72 microg/ml and > or =0.15 microg/ml was achieved in >98% of the children with at least 79% reaching anti-PRP concentrations of > or =1.0 microg/ml. Concentrations of > or =1.25 microg of Hib-tetanus vaccine produced mean anti-PRP responses at 18 weeks of > or =8.63 microg/ml and > or =0.15 microg/ml was achieved in 100% of the children with at least 88.9% reaching anti-PRP concentrations of > or =1.0 microg/ml. While mean antibody concentrations after either vaccine decreased over time, the proportion of children with antibody levels of > or =0.15 microg/ml had not changed significantly at the 9 month measurement. Immunologic memory was demonstrated by significant increases in mean antibody concentrations one week after the booster dose for doses > or =1.25 microg of HibCRM197 and Hib-tetanus to mean concentrations > or =37.71 and 16.07 microg/ml, respectively. There were no differences in antibody responses for vaccine doses > or =1.25 microg of the same vaccine or between the same concentrations of the two different vaccines. Our data suggest that doses of these vaccines of > or =1.25 microg may be sufficient to stimulate an immune response that offers both short and longer term protection from invasive Hib disease. PMID- 15542206 TI - Molecular analysis of Helicobacter pylori-associated gastric inflammation in naive versus previously immunized mice. AB - To identify mechanisms of immunity against Helicobacter pylori, we performed microarray analysis on gastric tissue from infected mice and mice vaccinated prior to challenge. RNA from gastric tissue was used to screen over 10,000 genes. MHC antigens and GTP binding proteins were upregulated in both groups. Infected mice were characterized by expression of innate host defense markers while immune mice expressed many IFN-gamma response genes and T cell markers. Results were confirmed for several genes by RT-PCR. CD4+ spleen cells from immune mice produced significantly more IFN-gamma than from infected mice. These results support a role for T cell regulated inflammation in H. pylori immunity. PMID- 15542207 TI - DNA and low titer, helper-free, recombinant AAV prime-boost vaccination for cytomegalovirus induces an immune response to CMV-pp65 and CMV-IE1 in transgenic HLA A*0201 mice. AB - A prime-boost immunization regimen allowed the use of low titer, helper-free rAAV pp65mII and rAAV-IE1 virus to elicit specific humoral and cellular responses to two important cytomegalovirus (CMV) antigens: the immediate-early 1 (IE-1) and pp65 proteins. Simultaneous immunization of both CMV proteins, using DNA vaccine priming followed by rAAV boost, induced antibody (Ab) response, CD8 lymphocytes with cytotoxic function, and detectible binding of the cognate peptide epitopes for human HLA A*0201 restriction using tetramer technology. PMID- 15542208 TI - Vaccination with gp120-depleted HIV-1 plus immunostimulatory CpG oligodeoxynucleotides in incomplete Freund's adjuvant stimulates cellular and humoral immunity in rhesus macaques. AB - Whole killed human immunodeficiency virus type 1 (HIV-1) immunogens contain the more conserved epitopes of HIV-1 and therefore may provide some utility as potential HIV-1 vaccine candidates. Previous studies have shown that synthetic oligodeoxynucleotides (ODN) containing unmethylated cytosine-guanine (CpG) dinucleotides trigger rapid stimulation of both CD4+ and CD8+ T cells. Here, we investigated whether immunization of rhesus macaques with an inactivated gp120 depleted HIV-1 immunogen, emulsified in incomplete Freund's adjuvant (IFA) together with immunostimulatory CpG-containing ODN (ODN 2006), would elicit HIV specific cellular and humoral immune responses. High titer anti-p24 antibody levels were induced in all four immunized animals that were sustained 6 weeks after the fifth and final boost at 23 months. These anti-gag antibodies mapped to linear B-cell epitopes within the matrix (MA), capsid (CA), p2, nucleocapsid (NC) and p6 proteins of HIV-1 gag. HIV-specific interferon-gamma-producing CD4+ and CD8+ T-cell responses were measured before and after the fourth and fifth immunizations by both intracellular cytokine (ICC) and ELISPOT techniques; responses were detected in three of the four immunized animals. CD4+ T-cell epitopes appear to map within amino acids 261-290 and 291-320 of p24 CA protein. Immunizations were well tolerated both locally and systemically. Based on these results, further studies of this approach are warranted. PMID- 15542209 TI - Immunological properties of FMDV-gP64 fusion proteins expressed on SF9 cell and baculovirus surfaces. AB - In the present report, we characterized the immune response and the protection conferred by recombinant baculoviruses or infected insect cells expressing the fusions gp64-P1 and gp64-site A FMDV antigens. Mice, vaccinated intraperitoneally with gp64-P1 immunogens, showed a low-antibody response and a variable degree of protection. However, when mice received recombinant baculoviruses or infected insect cells expressing the fusion protein gp64-site A, high-ELISA and seroneutralizing titers (SNT) against FMDV were elicited. All mice immunized with Sf9 cells expressing FMDV site A developed a protective immune response against challenge with virulent FMDV, indicating that the baculovirus display of foreign epitopes is a promising approach to biosynthetic vaccines. PMID- 15542210 TI - Editorial: an age of enlightenment or information overload? PMID- 15542211 TI - Early active mobilization of primary repairs of the flexor pollicis longus tendon with two Kessler two-strand core sutures and a strengthened circumferential suture. AB - This study reports our treatment of divided flexor pollicis longus (FPL) tendons by primary repair from 1999 to 2002. Forty-eight FPL repairs were performed using two Kessler two-strand repairs with a cross-linked Silfverskiold circumferential suture. All were rehabilitated by early active mobilization. Excellent or good results were observed in 73/77% of cases (White/Buck-Gramcko assessments, respectively). No patients (0%) ruptured their repair as a result of early active mobilization. Two patients (4%) developed post-operative infections with wound and tendon dehiscence. This combination of repairs addresses the problem of rupture of FPL during early mobilization which we experienced in previous studies. Its problems and alternatives are discussed. PMID- 15542212 TI - A comparative analysis of the biomechanical behaviour of five flexor tendon core sutures. AB - Five core suture techniques were compared by static tensile testing in vitro. Fifty porcine tendons were used. The core sutures were performed with 3-0 or 4-0 braided polyester suture (Ticron) and the over-and-over running peripheral sutures with 6-0 monofilament polypropylene (Prolene). The core sutures were: (1) Pennington modified Kessler (3-0), (2) Double Pennington modified Kessler (3-0), (3) 4-strand Savage (3-0), (4) 4-strand Savage (4-0), and (5) 6-strand Savage (4 0). Repairs were compared as paired in regard to one variable: the number of core suture strands, the suture calibre, or the suture configuration. Biomechanical differences between the repair groups started during the linear region, with the yield force and stiffness increasing along with the number of core suture strands. All three variables influenced the strain at the yield point. Thus, the strength of the intact repair can be improved by modifying the core suture. In all repairs gap formation started near the yield point after failure of the peripheral suture. The yield force represents the strength of the intact repair composite and should be considered the strength of the tendon repair. PMID- 15542213 TI - The non-opposable triphalangeal thumb: a new technique of management. AB - Fifteen non-opposable triphalangeal thumbs (eight patients) were treated with a new two-staged surgical technique. The first stage included excision of the proximal interphalangeal joint and osteotomy of the thumb metacarpal, and aimed to restore a correct thumb length, position and alignment and to create a wide thumb web space. The second stage consisted of an opponensplasty with refining of any of the first stage components, as necessary. The mean age at surgery was 3.2 years (range, 2.5-6 years). After a mean follow-up period of 42 months (range, 26 months to 7 years), the targets of treatment had been accomplished without loss of growth potential, joint stability or thumb motion. All children had improved function and cosmesis of the hand. PMID- 15542214 TI - Rehabilitation of digital nerve repair: is splinting necessary? AB - Forty isolated, sharp digital nerve divisions, which had been repaired by microsurgical techniques, were reviewed between 12 and 36 months postoperatively (mean, 20 months). Half of the repairs had been splinted beyond the immediate postoperative period and half had not. Non-splinted patients returned to work significantly quicker than those who were splinted. Splinted patients reported more stiffness and cold intolerance but splinting made no difference to either the measured sensibility or movement of the digit. We would conclude that, after repair of sharp, uncomplicated digital nerve divisions, splinting beyond the immediate postoperative period is at least unnecessary and may be deleterious. PMID- 15542215 TI - Sensory nerves transfers. AB - Various types of sensory nerve transfer in the hand are presented. They were all performed after brachial plexus injury to restore sensation, mainly to the adjacent borders of the thumb and index finger or to the ulnar aspect of the little finger. Twenty one cases were assessed. Their outcomes according to the Highet-Zachary system were: S2 in nine cases, S2 in four cases, S1 in four cases and S0 in four cases. PMID- 15542216 TI - Treatment of cubital tunnel syndrome by frontal partial medial epicondylectomy. A retrospective series of 55 cases. AB - The outcomes of 55 cases of cubital tunnel syndrome treated by a partial frontal epicondylectomy are presented at a mean follow-up of 38 months follow-up. According to McGowan classification, 25 cases were grade I (45%), 12 grade II (22%) and 18 grade III (33%). The results (Wilson and Krout classification) were excellent or good in 41 patients (75%), fair in nine patients and unchanged in five, without any worsening or recurrence. Total relief was reported in 80% of grade I, 75% of grade II and 66% of grade III patients. Seven painful scars and one persistent 15( composite function) elbow extension deficit were the only complications. The satisfaction rate was 93%. This technique preserves bony protection, the blood supply and gliding tissues for the nerve and nerve recovery were comparable to other surgical procedures. Residual pain at the osteotomy site was not a serious problem. PMID- 15542217 TI - Return-to-work interval and surgery for carpal tunnel syndrome. Results of a prospective series of 233 patients. AB - AIM OF THE STUDY: To evaluate the connection between the type of patient insurance and the time taken to return to work after carpal tunnel surgery. PATIENTS AND METHODS: Two hundred and thirty-three patients in full-time work were operated on for carpal tunnel syndrome between 1 January and 30 June 1998. They were divided into three groups: independent workers (n=87), wage earners in the private sector (n=90) and civil servants (n=56). Four categories were defined: manual workers, non-manual workers, patients with social security insurance and patients with workers compensation. The average return-to-work interval after surgery for each of the groups was evaluated and compared group by group. RESULTS: For independent workers the average time off work is 17 days, for those in the private sector it is 35 days, and for civil servants it is 56 days. Patients with social security insurance were off work for 32 days and those with workers compensation for 49 days. DISCUSSION: The comparison shows significant differences with regard to social security insurance: the return-to-work interval in civil servants is larger than for private sector workers, and this is higher than in independent workers. The difference between patients with workers compensation and those with social security insurance is 17 days and significant. There is a significant difference between manual and non-manual workers in independent and private sector workers. There is no significant difference between the sub-groups in the civil servants. These cross references enable us to work out the influence that social security status has on the return-to-work time following surgery. PMID- 15542218 TI - Self-administered hand symptom diagram for carpal tunnel syndrome diagnosis. AB - In a prospective case series in Singapore we studied whether a hand symptom diagram can be used to predict carpal tunnel syndrome as defined by standardized clinical and neurophysiological protocol. Forty patients (mean age, 48 years, range 18-76) with carpal tunnel syndrome found it difficult to complete the hand diagram, and only four patients were able to mark it helpfully. There was no relation between drawings, clinical and neurophysiological parameters. A self administered hand symptoms diagram is not useful for supporting a diagnosis of carpal tunnel syndrome amongst Singaporeans. Cultural factors may significantly influence subjective assessment scales such as the self-administered hand symptom diagram, which has been found effective in North American populations. PMID- 15542219 TI - Providing care for hand disorders a re-appraisal of need. AB - An audit of hand surgery activity in Derby during the period 1989-1990 produced manpower and resource recommendations for the speciality per 100,000 of population per year for the United Kingdom. The decade that followed the audit has seen major changes in health care provision, including reduced service activity by trainee doctors through restricted hours of work and less unsupervised surgery. A further audit of hand surgery activity was performed during 2000-2001 to assess the effects of these and other changes. This showed that there has been a 2% rise in trauma attendances, though trauma bed utilization had reduced by 12% and surgery time by 38%. Trauma out-patient visits had also reduced by 11%. Day-case trauma surgery rates were virtually unchanged at 63%. Women attend more frequently with traumatic hand injuries than they did 10 years ago and there is a rising incidence of hand injuries in the home, with a falling incidence at work. Elective referrals have risen by 36% and operations by 34%. The top ten diagnoses relate to the same conditions although their rankings have changed. Elective day-case surgery rates have risen from 64% to 94% over the decade. The 34% increase in elective operations has been absorbed within a 5% reduction in elective bed use and a 23% reduction in surgery time. Elective out patient visits have also dropped 14% overall. This audit indicates that in 2000 2001 one whole time equivalent hand surgeon can service a population of 125,000. The national requirement for a 56 million population would be 448 whole time equivalent hand surgeons. PMID- 15542220 TI - Scaphotrapeziotrapezoid (STT)-arthrodesis in Kienbock's disease. AB - This study reviews the results of 59 of 84 patients with severe Kienbock's disease who were treated with STT fusion. The average follow-up period was 4 (ranges: 2-8 years). The average arc of wrist extension and flexion was 67 degrees (60% of the contralateral side, 81% of pre-operative range) and that of ulnar and radial deviation was 31 degrees (52% of the contralateral side, 56% of pre-operative range). Pre-operative pain values (VAS) were 56 (non-stress) and 87 (stress) and were significantly higher than the postoperative values of 12 (non stress) and 41 (stress). Grip strength improved from 45kPa pre-operatively to 52kPa postoperatively. The mean modified Mayo wrist score was 63 points. The patients reported low disability in the DASH scores, with an average of 28 points. Our data show that STT fusion is a reliable and effective treatment for pain relief and offers a good functional result in advanced stages of Kienbock's disease. However the long-term effect of this procedure on radioscaphoid and other intercarpal joints is yet to be determined. PMID- 15542221 TI - Late reconstruction of neglected metacarpal shaft defects due to gunshot wound. AB - We evaluated eight patients after delayed treatment of nine metacarpal bone defects due to gunshot injuries. The mean length of the metacarpal defects was 3 cm and the average time between the gunshot injury and the reconstruction surgery was 10 months. Although all of the patients had been treated with wound irrigation and debridement immediately following injury, no attempt had been made to repair the metacarpal defect or to maintain metacarpal length. As a result, serious shortening had occurred. After the original length of the metacarpal had been restored by distraction of the soft tissues (1 mm/day), a tri-cortical iliac bone graft was inserted into the bone defect. The average follow-up time was 15 months. Clinical and radiological union was established in all cases after an average of 12 weeks. The mean grip strength of the hand and the mean range of motion of the metacarpophalangeal joint increased by 24% and 60%, respectively. PMID- 15542222 TI - Use of bioabsorbable osteofixation devices in the hand. AB - Bioabsorbable internal fixation by means of pins, tacks, screws and miniplates offers an alternative to metallic osteofixation for the stabilization of small bone fractures, osteotomies, ligament injuries and fusions in the hand. The advantages of using them include avoidance of metallic-implant-related long-term complications and a secondary removal operation. Currently the most commonly used devices are made of poly L-lactide (PLLA) and copolymers of polylactides (P(L/DL)LA) and polyglycolide (PLGA). In areas of mechanical stress, the use of ultra-high-strength self-reinforced devices is recommended. Biomechanical studies on fresh frozen bones have shown that the fixation rigidity achieved with self reinforced devices approaches that of metallic osteofixation methods. The reliability of modern implants has been confirmed in several experimental and clinical studies. PMID- 15542223 TI - Surgery of the hand in severe systemic sclerosis. AB - Ten patients with scleroderma and severe hand problems required surgery, and seven were available for follow-up (two died from scleroderma-related complications and one was lost to follow-up). The mean duration of follow-up was 4 (range 1.5-9) years. Thirty-three procedures were carried out, including five metacarpophalangeal joint excisional arthroplasties, 13 proximal interphalangeal joint fusions, ten distal interphalangeal joint fusions, and one thumb interphalangeal joint fusion. The metacarpophalangeal joint excision arthroplasties and proximal interphalangeal joint fusions were performed for the correction of severe fixed "finger-in-palm" deformities. Lesions of cutaneous calcinosis were removed in four patients. Fixation was satisfactory in all cases of interphalangeal joint fusion, with no cases of nonunion. Wound healing was satisfactory in six of seven patients. A second surgical procedure was required in three patients for the removal of tension band wires following interphalangeal fusion. Calcinosis was effectively removed using a high-speed dental burr. The results of hand surgery for systemic sclerosis are reliable, but goals must be limited and patient expectations should be modest. PMID- 15542224 TI - The effect of cellular proliferative activity on recurrence and local tumour extent of localized giant cell tumour of tendon sheath. AB - This study investigates whether the proliferative activity of giant cell tumour of tendon sheath is related to its recurrence rate and local aggressiveness. The clinicopathological and immunohistochemical features of 30 localized giant cell tumours of tendon sheath were studied and the influence of the MIB-1 staining index on recurrence, tumour extent around the phalanx and involvement of the bone were evaluated. No significant difference in the MIB-1 staining index was found between the lesions which recurred and those which did not. Also there was no significant association between local aggressiveness and the MIB-1 staining index. These results suggest that the proliferative activity of localized giant cell tumour of tendon sheath is not related to its high recurrence rate and local aggressiveness. PMID- 15542225 TI - The Sauve-Kapandji procedure and the Darrach procedure for distal radio-ulnar joint dysfunction after Colles' fracture. AB - This retrospective study evaluated the results of the Darrach procedure and the Sauve-Kapandji procedure for the treatment of distal radio-ulnar joint derangement following malunion of dorsally displaced, unstable, intraarticular fractures of the distal radius in patients under 50 years of age. Twelve of 18 possible patients in the Sauve-Kapandji group completed the disabilities of the arm, shoulder, and hand survey at a mean of 4 years postoperatively and nine of the 18 returned for a follow-up examination at a mean of 2 years. Twenty-one of 30 possible patients in the Darrach group completed the disabilities of the arm, shoulder, and hand survey at a mean of 6 years postoperatively and 13 of these 30 returned for follow-up examination at a mean of 4 years. The Darrach procedure and the Sauve-Kapandji procedure yielded comparable and unpredictable results with respect to both subjective and objective parameters. PMID- 15542226 TI - Hypoplastic synpolydactyly as a new clinical subgroup of synpolydactyly. AB - A large kindred which was first described in 1995 was investigated again. We present the clinical, radiological, genetic and surgical findings of the hand deformities found in homozygote individuals which we called "hypoplastic synpolydactyly". There were 125 affected (heterozygote or homozygote) people out of 245 subjects in the five last generations. We identified seven marriages of two affected people. Twelve offsprings, of these marriages had a homozygote genetic pattern and "hypoplastic synpolydactyly". From both the clinical and surgical perspectives, their hand deformity was distinctive from that of their parents. We surgically treated both hands of three individuals with this deformity. The hand deformity of these homozygotes was so complicated and distinctive that it can be evaluated as a new subgroup of synpolydactyly. PMID- 15542227 TI - The Consolidated Standards of Reporting Trials (CONSORT): better presentation of surgical trials in the Journal of Hand Surgery. AB - To assure readers that study results are scientifically valid, the methods of a clinical trial should be described adequately. Since randomization, blinding, and intention-to-treat-analysis are major bias-reducing techniques, these aspects should be reported most accurately. The Consolidated standards of reporting trials (CONSORT) are recommendations to improve the reporting of trials. CONSORT requires that trial authors describe basic methodological aspects that readers need to appraise the strengths of reported clinical trials. This article presents the CONSORT recommendations and explains some of their main aspects. From now on, the Journal of Hand Surgery will use CONSORT to assist authors of randomized controlled trials in improving the description of their studies. We believe that this decision increases the scientific validity of study reports and helps readers when critically appraising articles. PMID- 15542228 TI - Duplicate publication in the Journal of Hand Surgery. AB - This study evaluates the extent of duplicate publication in the Hand Surgery literature. A retrospective review of original articles published in the American and the British & European editions of Journal of Hand Surgery during the years 1999 and 2000 was performed using MEDLINE (PUBMED) search engine. Index articles suspected of dual publication were identified by using key words in the title and the names of the first, second and last authors. The full initial text was carefully studied and suspected duplicate articles were classified as dual, potentially dual or fragmented. Six hundred articles were evaluated, of which 25 (4%) index articles were identified with 33 "suspects". Eleven "index" and 15 "suspected" articles were cleared on closer scrutiny. Thus 14 "index" articles (2%) were found to be associated with 18 duplicated articles. Of these, four were classified as dual, five as potentially dual and nine as fragmented. We conclude that although duplicate publication of articles in the Journals of Hand Surgery (American and British/European Volumes) does occur, the incidence in the sample studied is lower than some other surgical journals. PMID- 15542229 TI - An isolated ring finger metacarpal shaft fracture?--beware an associated little finger carpometacarpal joint dislocation. AB - Dislocations of the ulnar carpometacarpal joint are easily missed because of a low index of suspicion as well as their subtle clinical and radiological features. Often, the presence of a more obvious adjacent injury also draws attention away from the carpometacarpal joint. Two cases of ring finger metacarpal shaft fractures with associated little finger carpometacarpal joint dislocations are presented. In both cases, the metacarpal fractures were diagnosed but the carpometacarpal joint dislocations were initially overlooked. The presence of an apparently isolated ring finger metacarpal fracture due to an indirect force should raise the possibility of an associated carpometacarpal joint injury. PMID- 15542230 TI - Closed multiple tendinous rupture of flexors and extensors of hand: a case report. AB - Multiple ruptures of the extensor and flexor tendons of the fingers, thumb and wrist at the musculotendinous junctions are reported after a blast injury. PMID- 15542231 TI - The use of chloramphenicol ointment as an adhesive for replacement of the nail plate after simple nail bed repairs. AB - Fingertip injuries are common and are often repaired. The nail plate is replaced to protect the repair and maintain the alignment of the nail bed edges after repair. Once replaced, this can be secured by any means and often a suture is used. We advocate that a small bleb of chloramphenicol provides a simple method of securing the nail plate. It is fast, easy and secure. In addition it provides an antibacterial effect, which may be of some benefit. PMID- 15542232 TI - Dorsal Dupuytren's disease causing a swan-neck deformity. AB - A previously undescribed lesion of Dupuytren's disease is presented. An oblique cord coursed parallel to the oblique retinacular ligament of Landsmeer, but inserted proximal to the proximal interphalangeal joint, tethering the central slip and radial lateral band across the intervening transverse retinacular ligament. Contraction of this cord caused a rigid swan-neck deformity. Excision of the cord resulted in complete resolution of the deformity and a full range of motion in the affected digit. PMID- 15542234 TI - Re. Prevalence and epidemiological variation of anomalous muscles at Guyon's canal. PMID- 15542235 TI - Re. A model for the conservative treatment of mallet finger. PMID- 15542236 TI - Inhibitory effects of first syllable-frequency in lexical decision: an event related potential study. AB - Electrophysiological correlates of the behaviorally well-documented inhibitory effect of first syllable-frequency during lexical access are presented. In a lexical decision task, response times to words with high-frequency first syllables were longer than those to words with low-frequency first syllables and resulted in more negative event-related potentials (ERPs) in an early time window from 190 ms to 280 ms and in the N400 component. The onset of the observed first syllable-frequency effect was prior to the onset of the effect of lexicality (i.e., the first reliable differentiation in ERP waveforms in response to words and pseudowords, a potential marker of lexical access). The present study's results support Barber et al.'s [Neuroreport 15 (2004) 545] notion of the prelexical nature of the first syllable-frequency effect by (A) providing evidence for electrophysiological correlates of first syllable-frequency in another, non-Romance orthography (i.e., German), (B) relating the onset of the first syllable-frequency effect to the onset of the lexicality effect and (C) strengthening this pattern of results by means of a novel item-based analysis of ERP data. Implications of the prelexical nature of the inhibitory first syllable frequency effect for computational models of reading, specifically for Ans et al.'s [Psychol. Rev. 105 (1998) 678] multiple-trace memory (MTM) model of reading are discussed. PMID- 15542237 TI - Bone marrow-derived mast cells in mice respond in co-culture to scorpion venom activation of superior cervical ganglion neurites according to level of expression of NK-1 receptors. AB - In virtually all tissues of the body, mast cells are closely associated with nerve fibers, mostly of sensory origin. While mast cells can be activated by substance P, evidence for the involvement of NK-1 receptors is very limited. To study functional interactions between mast cells and peripheral nerves, bone marrow-derived mast cells (BMMC) and superior cervical ganglia (SCG) were co cultured. Murine bone marrow-derived mast cells are homologues for mucosal mast cells and have recently been shown to express NK-1 receptors. Bi-directional interaction was studied using a fluorescent calcium indicator as an index of cellular activation. Scorpion venom, not affecting BMMC by itself, caused a rapid increase in neurite fluorescence subsequently followed by activation of the mast cell. The latter was inhibited by the NK-1 receptor antagonist SR140333, showing the direct involvement of substance P and its receptor in this co-culture system. Activation of BMMC seemed to be directly correlated with extent of NK-1 receptor expression. Immature c-kit positive cells not expressing NK-1 gave a negligible response to neurite activation. In addition, there was a maximum stimulation occurring when NK-1 expression exceeded 16% on BMMC after cytokine stimulation. Our findings show that the expression of NK-1 receptors appears to be important for nerve-mast cell communication. PMID- 15542238 TI - Selective translocation of diacylglycerol kinase zeta in hippocampal neurons under transient forebrain ischemia. AB - The molecular mechanisms responsible for differential neuronal vulnerability to ischemic injury are incompletely understood. Previous studies have reported that the expression and activity of protein kinase C (PKC), some subtypes of which are activated by Ca(2+) and diacylglycerol (DG), are altered after ischemic insults. Therefore, DG kinase (DGK), which is responsible for controlling PKC activity through DG metabolism, may also be involved in this process. DGKzeta, which is abundantly expressed in the brain, contains a nuclear localization signal (NLS), suggesting its involvement in some nuclear processes in neuronal cells. To elucidate the functional implications of DGKzeta in ischemia, we examined detailed localization of DGKzeta in rat brain after ischemic insults. We used an ischemic model of global cerebral ischemia for 20 min by bilateral common carotid artery occlusion combined with hypotension and followed time-points of reperfusion. DGKzeta expression was evaluated by immunohistochemistry using affinity-purified anti-DGKzeta antibody. In sham-operated rats, a strong DGKzeta immunoreactivity was observed in the nucleus of neurons in various parts of the brain. In the global ischemic model DGKzeta-immunoreactivity was reduced in intensity in the hippocampal formation and detected in the cytoplasm of CA1 pyramidal neurons throughout reperfusion time courses. Change in the subcellular localization was restricted to the pyramidal cells in CA1 and later in CA3, but not observed in other areas of hippocampus. No change was observed in the cerebral and cerebellar cortices. The present study suggests that DGKzeta might be involved in the process of selective vulnerability of hippocampal pyramidal neurons in postischemic brain. PMID- 15542239 TI - A deletion in DJ-1 and the risk of dementia--a population-based survey. AB - The DJ-1 gene is associated with autosomal recessive early-onset Parkinsonism, most likely through its role in defense against oxidative stress. Oxidative stress is not only involved in Parkinson's disease, but also in other neurodegenerative disorders, such as dementia. We assessed the presence of a 14 kb DJ-1 deletion in 191 patients with dementia, ascertained from the genetically isolated population where the first kindred with DJ-1 related Parkinsonism was originally identified. The control group consisted of 129 non-demented subjects. We found the deletion in two patients and one control. There was no evidence for an increased risk of dementia in carriers. All subjects were heterozygous for the deletion and related to a common ancestor within eight generations. Our results suggest it is unlikely that haploinsufficiency in the DJ-1 gene imparts an increased risk for dementia. PMID- 15542240 TI - Transmission disequilibrium analysis of the GSN gene in a cohort of family trios with schizophrenia. AB - Apoptosis is thought to play a role in neuronal pathology in schizophrenia. Recently, the GSN gene was reported to have anti-apoptotic properties. In a genome-wide expression analysis on schizophrenia, GSN was also found to be significantly down-regulated in schizophrenia. All the hints suggest that GSN is a novel candidate gene in occurrence of schizophrenia. In this work, we genotyped 3 SNPs around the GSN locus in 493 sets of the Han Chinese trio sample using allele-specific PCR. A weak association or a marginally positive result was detected (0.05 for P-value of the overtransmitted haplotype and 0.02 for a global P-value). PMID- 15542241 TI - Brain activation to phobia-related words in phobic subjects. AB - Behavioural studies suggest that phobic subjects are hypersensitive in the processing of phobia-related linguistic stimuli. We used functional magnetic resonance imaging (fMRI) to investigate blood oxygen level dependent (BOLD) brain activation to phobia-relevant words in spider phobic and non-phobic subjects. Phobia-related versus phobia-unrelated words elicited increased activation in prefrontal cortex, insula, and posterior cingulate cortex in spider phobics, while these effects were absent in controls. Furthermore, between-group comparisons confirmed that differential activations within these brain regions were specifically due to increased responses to phobia-related stimuli in phobics. Our results provide first insights into brain activation patterns when phobics are confronted with phobia-specific linguistic information und suggest a neural network for the processing of these threatening stimuli. PMID- 15542242 TI - Analysis of the autism chromosome 2 linkage region: GAD1 and other candidate genes. AB - Autism has a strong and complex genetic component, involving several genes. Genomic screens, including our own, have shown suggestive evidence for linkage over a 20-30 cM region on chromosome 2q31-q33. Two subsequent reports showed that the linkage evidence increased in the subset of families with phrase speech delay (PSD), defined as onset of phrase speech later than 3 years of age. To further investigate the linkage in the presumptive candidate region, microsatellite markers in a 2 cM grid covering the interval from 164 to 203 cM were analyzed in 110 multiplex (2 or more sampled autism patients) families. A maximum heterogeneity LOD (HLOD) score of 1.54 was detected at D2S1776 (173 cM) in the overall dataset (dominant model), increasing to 1.71 in the PSD subset. While not conclusive, these data continue to provide suggestive evidence for linkage, particularly considering replication by multiple independent groups. Positive LOD scores extended over the entire region, continuing to define a broad candidate interval. Association studies were performed on several functional candidates mapping within the region. These included GAD1, encoding GAD67, whose levels are reduced in autopsy brain material from autistic subjects, and STK17B, ABI2, CTLA4, CD28, NEUROD1, PDE1A, HOXD1 and DLX2. We found no evidence for significant allelic association between autism and any of these candidates, suggesting that they do not play a major role in the genetics of autism or that substantial allelic heterogeneity at any one of these loci dilutes potential disease-allele association. PMID- 15542243 TI - Wallerian degeneration after crush injury of rat sciatic nerve increases endo- and epineurial tumor necrosis factor-alpha protein. AB - The pro-inflammatory cytokine tumor necrosis factor-alpha (TNF) contributes to injury-induced peripheral nerve pathology and to the development of neuropathic pain. Here, we investigated whether TNF protein is altered at the site of crush injury of rat sciatic nerve using enzyme-linked immunoassay (ELISA) and immunohistochemistry (IHC). TNF protein levels determined by ELISA were low in nerve homogenates from naive rats. After crush injury, local TNF increased rapidly with a two-fold increase on day 0.5. TNF content remained elevated on day 3 and returned to baseline levels again by day 14 after crush. IHC revealed prominent TNF-immunoreactivity in many epineurial macrophages on days 0.5 to 3 after crush injury. These data indicate that TNF protein is early and transiently upregulated at the site of peripheral nerve trauma. PMID- 15542244 TI - Involvement of alpha6/alpha3 neuronal nicotinic acetylcholine receptors in neuropsychiatric features of Dementia with Lewy bodies: [(125)I]-alpha-conotoxin MII binding in the thalamus and striatum. AB - Dementia with Lewy bodies (DLB) is a neurodegenerative disease associated with a range of neuropsychiatric symptoms and reduced expression of neuronal nicotinic acetylcholine receptors (nAChRs) in neocortex, hippocampus, thalamus and basal ganglia. To determine whether there are selective associations between alterations in alpha6/alpha3 neuronal nicotinic acetylcholine receptors (nAChRs) and the two key neuropsychiatric features of DLB, impaired consciousness (IC) and visual hallucinations (VH), quantitative [(125)I]-alpha-conotoxin MII ([(125)I] alpha-Ctx MII) autoradiography was undertaken on 28 people with DLB and 15 control cases from the Newcastle Brain Bank. There was a highly significant overall trend for reduced thalamic [(125)I]-alpha-Ctx MII binding in DLB (p < 0.001), with significant deficits in the centromedian, ventral lateral and ventroposterior medial thalamic nuclei (p < 0.05), together with caudate and putamen (p < 0.001). [(125)I]-alpha-Ctx MII binding was significantly lower in DLB cases with IC than without IC in the putamen (p < 0.05), however there was no significant association between [(125)I]-alpha-Ctx MII binding and VH. Reductions in [(125)I]-alpha-Ctx MII binding in caudate and putamen were paralleled by similar reductions in [(125)I]PE2I binding. [(125)I]PE2I binding was also significantly lower in DLB cases with IC than without IC in the caudate (p < 0.05) and putamen (p < 0.001). These results demonstrate that deficits in alpha6/alpha3 nAChRs occur in specific brain regions in DLB, may in part be related to the loss of dopaminergic neurons and may contribute to the development of impaired consciousness in the disorder. PMID- 15542245 TI - Genetic association study of PINK1 coding polymorphisms in Parkinson's disease. AB - Parkinson's disease (PD) is the most common neurodegenerative movement disorder with a substantial genetic component (which is more pronounced in earlier onset cases). In addition to three well-confirmed PD genes (SNCA, parkin and DJ-1), mutations in the PTEN Induced Kinase (PINK1) gene have recently been identified in families with recessive early onset PD. We tested the hypothesis that three common coding variations (Leu63Leu, Ala340Thr and Asn521Thr) could increase the risk of PD. We performed a case control association study in a series of 91 PD cases (Caucasian of Canadian origin) and 182 normal controls. The patients were largely pre-selected for having an early age of onset (<50 years) and/or a positive family history. Our results did not reveal any evidence of association between PD and any of the three SNPs at the allelic or genotypic levels (p > 0.25). Furthermore, we did not detect a modifying effect for any genotype upon the age of onset in the PD group (p > 0.19). Nevertheless, it remains to be evaluated whether PINK1 variations contribute to the risk of common late onset sporadic PD. PMID- 15542247 TI - Visual and motor constraints on trajectory planning in pointing movements. AB - The aim of the present study was to show that planning and controlling the trajectory of a pointing movement is influenced not solely by physical constraints but also by visual constraints. Subjects were required to point towards different targets located at 20 degrees , 40 degrees , 60 degrees and 80 degrees of eccentricity. Movements were either constrained (i.e. two-dimensional movements) or unconstrained (i.e. three-dimensional movements). Furthermore, movements were carried out either under a direct or a remote visual control (use of a video system). Results revealed that trajectories of constrained movements were nearly straight whatever the eccentricity of the target and the type of visual control. A different pattern was revealed for unconstrained movements. Indeed, under direct vision the trajectory curvature increased as the eccentricity augmented, whereas under indirect vision, trajectories remained nearly straight whatever the eccentricity of the target. Thus, movements controlled through a remote visual feedback appear to be planned in extrinsic space as constrained movements. PMID- 15542246 TI - Alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate receptor subunit expression in rat olfactory bulb. AB - The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors (AMPARs) mediate rapid responses at most central excitatory synapses, including those in the olfactory bulb (OB). These receptors are composed of the glutamate subunits GluR1-4, which each has two splice variant (flip/flop) forms. We recently showed that AMPARs on OB neurons are kinetically and pharmacologically diverse. Here, we explored whether this functional heterogeneity reflects a diverse expression of AMPAR subunits and/or splice variants. Total RNA from rat OBs was amplified by RT-PCR. Digestion of the panGluR PCR product with subunit specific restriction enzymes revealed that the OB expresses mRNAs for GluR1-4 but in different relative amounts i.e., GluR2 (61 +/- 2.4%), GluR1 (31 +/- 3.5%), GluR4 (6.3 +/- 1.4%), GluR3 (1.4 +/- 0.7%). Furthermore, GluR2 and GluR4 transcripts were composed of similar amounts of flip and flop, whereas GluR1 and GluR3 transcripts consisted mostly of flip. If similar to other brain regions, this heterogeneity in patterns of expression may facilitate information processing. PMID- 15542248 TI - Analysis of MDR1 haplotypes in Parkinson's disease in a white population. AB - The MDR1 multidrug transporter is important in regulating environmental xenobiotics and hence may play a causative role in Parkinson's disease (PD). MDR1 haplotype comprising 2677 G > T/A and 3435 C > T may be protective against PD. Using a case control methodology, we investigated the association of MDR1 haplotypes (single nucleotide polymorphisms (SNPs) 2677 G > T/A and 3435 C > T) in a Polish PD population. Seven SNPs, extending from the promoter to exon 28 of the MDR1 gene in 158 PD patients and 139 healthy controls were evaluated. Specifically we examined the association of haplotypes containing SNPs 2677 G > T/A and 3435 C > T and risk of PD. The multivariate logistic regression model was used to evaluate the effects of the covariates on the phenotypes. Haplotypes' frequencies were estimated using the Expectation-Maximization algorithm. The frequency of each individual SNPs; -41 A > G (intron -1), -145 C > G (exon 1), 129 T > C (exon 1), 1236 T > C (exon 12), 2677 G > T/A (exon 21), 3435 C > T (exon 26), and 4036 A > G (exon 28) did not differ between PD and controls. However, there was a trend towards significance in PD patients having the haplotype 2677G-3435C (p < 0.09, chi-square 2.85, odds ratio 0.25, 95% CI 0.06 1.08). Haplotype constructs of the other loci did not differ significantly between the two groups. There was a weak protective effect of the haplotype 2677G 3435C in our white population. However, the MDR1 haplotypes did not generally modulate the risk of PD. PMID- 15542249 TI - Short-term habituation of the intracranially recorded auditory evoked potentials P50 and N100. AB - At an interstimulus interval (ISI) of 500-ms stimulus repetition leads to a strong decrease in cortical response. The functional foundation of this response suppression (or sensory gating) is yet not fully understood. Experiments on short term habituation using the same stimulus material as sensory gating experiments and same ISI might help to elucidate the mechanisms behind the P50 suppression. Event-related potentials were recorded intracranially in epileptic patients undergoing presurgical evaluation with subdural and depth electrodes. Stimulus material consisted of trains of six clicks, with the last stimulus deviating in pitch and duration. P50 and N100 were calculated for each stimulus in the train separately and compared by analysis of variance (ANOVA). A highly significant amplitude reduction was found from the 1st to 2nd stimulus for both P50 and N100. From the 2nd to 5th stimulus no further amplitude decrease was observable. The deviating 6th stimulus led to a response recovery of both components, but the P50 elicited by the 6th stimulus was still smaller than the P50 of the 1st stimulus. Current results indicate that the P50 suppression as investigated in sensory gating experiments seems to be completed after the 2nd stimulus. PMID- 15542250 TI - Sexual dimorphism in the antinociception mediated by kappa opioid receptors in the rat temporomandibular joint. AB - This study assessed the effect of the kappa opioid receptor agonist U50,488 administered into the rat temporomandibular joint (TMJ) on nociceptive behavioral responses evoked by formalin injected into the same site. Groups consisted of females, stratified into proestrus and diestrus phases of the estrous cycle, and males. Intra-TMJ formalin induced significantly different dose-dependent responses among the three groups, with diestrus females showing greater responses than males or proestrus females; therefore, equi-nociceptive formalin doses were chosen to test the effects of U50,488. U50,488 significantly reduced formalin induced nociceptive behavior in all groups, but the reduction was significantly greater in females, especially those in diestrus. Pre-injection of the selective kappa opioid receptor antagonist nor-binaltorphimine (nor-BNI) into the same site significantly attenuated the effect of U50488; U50,488 injection into the contralateral TMJ failed to reduce nociceptive behavior. These findings support a role for kappa opioid receptors local to the site of inflammation to modulate inflammatory pain. Furthermore, since plasma levels of ovarian hormones are low during diestrus, these findings are consistent with the suggestion that sex hormones may play an antagonistic role in these peripheral kappa-mediated effects. PMID- 15542251 TI - Treadmill exercise improves short-term memory by suppressing ischemia-induced apoptosis of neuronal cells in gerbils. AB - In the present study, the effect of treadmill exercise on short-term memory, apoptotic neuronal cell death, and cell proliferation in the hippocampal dentate gyrus following transient global ischemia in gerbils was investigated. Step-down inhibitory avoidance task, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay, and immunohistochemistry for caspase-3 and 5 bromo-2'-deoxyuridine (BrdU) were used. Ischemia was induced by the occlusion of both common carotid arteries (CCA) of gerbils for 5 min. Gerbils in exercise groups were forced to run on a treadmill for 30 min once a day for 10 consecutive days. Such treadmill exercise improved short-term memory by suppressing the ischemia-induced apoptotic neuronal cell death in the dentate gyrus. In addition, treadmill running suppressed the ischemia-induced cell proliferation in the dentate gyrus. The present results suggest that treadmill exercise overcomes the ischemia-induced apoptotic neuronal cell death and thus facilitates the recovery following ischemic cerebral injury. PMID- 15542252 TI - Arginine release from rat cerebellar astrocytes: autocrine roles for glutamate and nitric oxide? AB - In this study we have investigated the relationship between glutamate and arginine release from cultured cerebellar astrocytes. We found that the nitric oxide (NO) donor S-nitroso-N-acetylpenicillamine (SNAP) promoted the release of both amino acids in a concentration-dependent manner, and that these responses were partially reversed by a guanylate cyclase inhibitor. Application of the non NMDA glutamate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) resulted in a 60% reduction in basal arginine release but no change in that of glutamate. This effect was not overcome by the subsequent addition of SNAP despite a two-fold increase in glutamate release. Incubation with the nitric oxide synthase (NOS) inhibitor N-nitro-L-arginine methyl ester (L-NAME) elicited 40 and 60% reductions in the basal release of glutamate and arginine, respectively. Basal release of both amino acids was restored by the addition of SNAP. We conclude that glutamate released from cerebellar astrocytes in response to increased levels of extracellular NO acts in an autocrine manner to promote arginine release via the activation of non-NMDA receptors. In addition, our data suggest that basal glutamate release is regulated to some extent by tonic NO synthesis in these cells. PMID- 15542253 TI - Clinicopathological correlates of aspartyl (asparaginyl) beta-hydroxylase over expression in cholangiocarcinoma. AB - Aspartyl (asparaginyl) beta-hydroxylase (AAH) expression in surgically resected intrahepatic cholangiocarcinoma significantly correlated with tumor size, growth type, differentiation, vascular invasion, and prognosis after surgery. AAH may have a role in regulating invasive or metastatic tumor cell growth of human intrahepatic cholangiocarcinoma. BACKGROUND: Recent studies demonstrated increased expression of the AAH gene in the majority of cholangiocarcinomas. The present study was undertaken to determine the relationship between high or low levels of AAH expression and the clinical course of intrahepatic cholangiocarcinoma (ICC). METHODS: AAH expression was examined in 50 surgically resected primary ICCs, 12 samples of normal liver, and 12 cases of primary sclerosing cholangitis (PSC). The sections were evaluated by immunohistochemical staining with the FB-50 monoclonal antibody to human AAH protein. The sections were examined under code and graded for relative levels of AAH immunoreactivity. The results were analyzed with respect to multiple clinical and histopathological variables to determine correlates of AAH expression in ICCs. RESULTS: Forty-six of the 50 (92%) ICCs had AAH immunoreactivity, whereas the 12 normal liver and 12 PSC specimens were AAH negative. In the ICC specimens, the highest levels of AAH immunoreactivity were detected at the infiltrating margins that interfaced with uninvolved liver tissue, and the lowest levels occurred in the central portions of the tumors. Multivariate analysis demonstrated that high levels of AAH expression were correlated with tumor size (P < 0.05), infiltrative growth pattern (P < 0.01), aggressive histological grade (P < 0.01), vascular invasion (P < 0.05), and poor prognosis (P < 0.05). CONCLUSIONS: These findings suggest that AAH has an important role in regulating invasive or metastatic tumor cell growth of human ICC, and that high levels of AAH expression correlate with poor prognosis. PMID- 15542254 TI - Correlation of elevated plasma levels of two structurally related growth factors, heparin affin regulatory peptide and midkine, in advanced solid tumor patients. AB - Heparin affin regulatory peptide (HARP) and midkine (MK) are growth factors, expressed in carcinomas, neuroblastomas and gliomas. In this study, we measured the levels of HARP and MK in plasma samples from 77 cancer patients. The patients had advanced tumors with loco-regional (n=18) or metastatic (n=49) diseases and 10 patients have their diseases limited to the primary site. HARP and MK plasma concentrations were significantly higher in all of these different subgroups of cancer patients (P<0.05 in all cases), when compared to healthy controls (n=30). Neither HARP nor MK levels were significantly different between patients with loco-regional and metastatic tumors (P=0.203 and 0.242, respectively). Moreover, a strong correlation between the elevations of the plasma levels of these two proteins (r2=0.546) in these cancer patients was found. Measurements of these secreted angiogenic growth factors may be useful for evaluation of cancer diagnosis. PMID- 15542255 TI - Immunosuppressive mechanisms in the microenvironment of malignant pleural effusions. AB - Malignant effusions in serous cavities constitute a unique milieu for direct contact of tumor cells with host lymphoid cells in a fluid phase. The aim of this study was to depict agents responsible for suppression of lymphoid cells with putative anti-tumor potential. Pleural effusions drawn from 44 (18 non-malignant and 26 malignant) patients were tested for selected cytokines--interleukin-10 (IL 10), transforming growth factor beta (TGF-beta1) and soluble Fas ligand (sFasL) and nuclear membrane proteins (NMPs) content by ELISA. TCR-zeta expression of T cells and TUNEL reaction for apoptosis were evaluated by three color flow cytometry. Both cytokine concentrations were found to be significantly elevated in malignant pleural effusions (MPE) as compared to non-malignant ones. It was also true for sFasL content. Moreover, NMPs corresponding to decoy cell fragments, were also heightened in MPE. Concentrations of NMPs correlated with the percent of apoptotic (TUNEL+) T CD3+ lymphocytes and inversely correlated with the percent of T cells. The low expression of TCR-zeta chain on T cells corresponded to high concentration of sFasL in MPE. In conclusion, the above data suggest that out of three suppression agents tested, only sFasL appears to show correlation with the downregulation of T cells in MPE. PMID- 15542256 TI - A recurrent chromosome 4 marker in primary squamous cell lung cancer. AB - A sample of eight primary lung tumors histologically identified as squamous cell type carcinomas (SQC) were cytogenetically analyzed after a short-term (7 days) culture. Several numerical and structural abnormalities were observed. As G banding analysis is unable to fully characterize complex marker chromosomes, we applied the fluorescence chromosome painting technique to identify one as a marker derived from a chromosome 4 that has not been described until now. We found it in half of the analyzed cases. PMID- 15542257 TI - Behavioural risk factors for cervical cancer from a prospective study in Khon Kaen, Northeast Thailand. AB - Fifty seven cases of cancer of the cervix were identified within a cohort of 16,648 women enrolled in a cohort study of lifestyle and cancer in a rural population of Northeast Thailand. They were compared with 228 matched controls from the same cohort, with respect to demographic, behavioural and reproductive factors potentially associated with risk of the disease. Number of pregnancies and age at having first child were associated with the risk of cervical cancer, but in contrast to studies elsewhere, age at having first sexual intercourse, number of sexual partners and sexually-transmitted infections were not associated with risk. The lack of effect may be simply due to the small size of the study. But it is also likely that these aspects of female sexual behaviour are not very relevant in a relatively conservative rural population. These aspects may be investigated further in future, as new incident cancers are accrued into the cohort. PMID- 15542258 TI - Survey of surgeons' and internists' knowledge of colorectal cancer screening. AB - BACKGROUND: Colorectal cancer (CRC) is the most common of the gastrointestinal cancers in Israel, with an incidence of 50 cases per 100,000 for men and 45 per 100,000 for women in 2000. AIM: The low rate of patient compliance noted in CRC screening program conducted in our center, prompted the present survey of the knowledge and practices regarding CRC screening recommendations. METHODS: A 23 item questionnaire formulated according to the Israeli Ministry of Health policy and recommendation of the American Gastroenterological Association was distributed among the physicians of the six departments of internal medicine, and the department of surgery of the Rabin Medical Center. The relative ratio of correct answers was calculated for every question and by subgroups: internists and surgeons, experts and residents in internal medicine and in surgery. RESULTS: Fifty-five of 80 physicians (69%) completed the questionnaire. The total score for all participants was 15.82 (average 0.68), significantly lower than the optimal correct score of 23 (average 1.00). The internists had a similar total score to the surgeons, 15.61 and 15.28. When the questions were clustered according to specific issues and only the correct answers for each cluster scored, the success rate decreased with a range of 19-69%. Only 22% of responders did not advise fecal occult blood test (FOBT) for the high-risk population; 57% thought the screening age range was 40-65 years, and only 37% knew that the high risk criteria include a family history of ovarian cancer. CONCLUSION: Both internists and surgeons' knowledge about screening recommendations of CRC can be improved. Health authorities should invest greater efforts in continuing physician education. We believe that this strategy will increase the participation rate in CRC preventive programs. PMID- 15542259 TI - The role of low-level magnification in visual inspection with acetic acid for the early detection of cervical neoplasia. AB - Several studies have investigated the accuracy of naked eye visual inspection with acetic acid (VIA) in the early detection of cervical neoplasia. It is not clear whether low-level (2-4x) magnification (VIAM) can improve the sensitivity and specificity of VIA. The accuracy of both VIA and VIAM, provided by independent health workers, were evaluated in three cross-sectional studies involving 18,675 women aged 25-65 years in Kolkata and Mumbai in India. All screened women were investigated with colposcopy and biopsies were obtained based on colposcopy findings. The final disease status was based on the reference standard of histology (if biopsies had been taken) or colposcopy. Data from the studies were pooled to calculate the test characteristics for the detection of high-grade squamous intraepithelial lesions (HSIL). 14.1% and 14.2% were positive on testing with VIA and VIAM respectively. Two hundred twenty-nine were diagnosed with HSIL and 68 with invasive cancer. The pooled sensitivity, specificity, positive and negative predictive values for VIA in detecting high-grade squamous intraepithelial lesions (HSIL) were 60.3% (95% CI: 53.6-66.7), 86.8% (95% CI: 86.3-87.3), 5.9% (95% CI: 5.0-7.0), and 99.4% (95% CI: 99.2-99.5), respectively. The values were 64.2% (95% CI: 57.6-70.4), 86.8% (95% CI: 86.2-87.3), 6.3% (95% CI: 5.3-7.3) and 99.4% (95% CI: 99.3-99.6), respectively, for VIAM. Low-level magnification did not improve the test performance of naked eye visualization of acetic acid impregnated uterine cervix. PMID- 15542260 TI - Evaluation of CaDet, a computer-based clinical decision support system for early cancer detection: a comparison with the performance of clinicians. AB - Based on an analysis of the epidemiologic and clinical attributes of individual patients, the CaDet computer program presents physicians with data patterns that may require clinical attention with regard to early cancer detection. This study evaluated the performance of the program as a possible tool to aid clinicians in the latter. Patient data were obtained by a questionnaire from 160 healthy volunteers. Scored cancer alerts generated by the computer program in response to this data were evaluated in comparison to similar alerts provided by five expert internists who reviewed the same information in blinded fashion. The alert profiles generated by the computer for each of the patients examined were highly correlated to those provided by the clinicians. The computer's alert rate increased with the number of physicians who raised corresponding alerts (20%, 32%, 54%, 73% and 91%, respectively, for 1, 2, 3, 4 and 5 physicians, p < 0.0001) and with the overall weight assigned by the physicians to these alerts (22%, 56%, 84% and 90% for cumulative scores of 1-4, 5-8, 9-12 and 13 or more points, respectively, p < 0.0001). It is concluded that the CaDet computer program may have a role in improving early detection, pending the results of further clinical research. PMID- 15542261 TI - Bilateral oophorectomy and breast cancer risk reduction among women with a family history. AB - PURPOSE: We investigated whether removal of the ovaries is an appropriate risk reduction option for women at elevated risk of breast cancer based on family history of breast cancer. PATIENTS AND METHODS: This question was investigated among a group of 851 women less than age 60 who underwent bilateral oophorectomy between 1970 and 1994 for various reasons. Questionnaire information was collected from 680 (80%) and women were grouped into family risk categories. Reported occurrences of breast cancer were compared to expected rates based on the Gail model. RESULTS: The number of observed breast cancers among women in the cohort was lower than expected for all levels of familial risk, with women in the highest risk groups experiencing about half to one-fourth the number of cancers expected. The apparent protective effect of oophorectomy was stronger among women who were both premenopausal and less than age 50 at time of surgery. CONCLUSION: These data support oophorectomy as a valid breast cancer prevention option for women of all risk levels. PMID- 15542262 TI - Influence of direct referrals on time to diagnosis after an abnormal breast screening result. AB - This study examined the influence of a direct referral process implemented by a population-based provincial breast screening program on the time from screening to first procedure, first procedure to diagnosis, and screening to diagnosis following an abnormal breast screening result. The direct referral process shifted the responsibility for the coordination of diagnostic follow-up procedures from the family physician to the screening program. Three cohorts of women were included: a control cohort (screened prior to the initiation of a direct referral process, n = 1347), a usual care cohort (screened after the initiation of a direct referral process but for whom permission to refer was denied by the family physician, n = 1225), and a direct referral cohort (screened after the initiation of the direct referral process and for whom permission to refer was given by the family physician, n = 1232). The direct referral cohort was subdivided into women referred to a breast health centre (BHC group) (n = 606) and women referred to a diagnostic facility (diagnostic group) (n = 626). The direct referral cohort completed all three time intervals significantly faster than the other two cohorts (P < 0.0001). The diagnostic group experienced a significantly lower time from screening to the first procedure than the other cohorts or the BHC group (P < 0.0001). However, the BHC group had a significantly lower time from first procedure to diagnosis than the other cohorts or the diagnostic group (P < 0.0001). The control and the usual care cohorts were not significantly different from each other (P = 0.6250). The direct referral process significantly reduced the time to diagnosis after an abnormal screening result. PMID- 15542263 TI - Glutathione S-transferase P1 Ile105Val polymorphism, cigarette smoking and prostate cancer. AB - The enzyme glutathione S-transferase P1 (GSTP1) detoxifies carcinogenic products of tobacco smoke. This exploratory case-control study evaluates the possible effect modification by the GSTP1 Ile105Val polymorphism (replacement of isoleucine by valine at codon 105) on smoking and prostate cancer. Because the Val variant possesses up to a five-fold greater enzymatic activity towards the carcinogenic metabolites of tobacco smoke, the Ile allele is expected to be related to an increase in the risk of prostate cancer among smokers. GSTP1 genotype and epidemiological data were obtained from 122 cases of prostate cancer and 135 healthy males as controls. A logistic regression model was used to estimate odds ratios and 95% confidence intervals. The adjusted OR of homozygous Ile compared to other genotypes for prostate cancer was 1.21 (95% CI: 0.61-2.83). Smoking was not significantly associated with prostate cancer with an adjusted OR of 1.56 (95% CI: 0.78-3.12). However, among individuals with the Ile/Ile genotype, smoking was strongly associated with an increased risk of prostate cancer with an adjusted odds ratio of 4.09 (95% CI: 1.25-13.35). A potential multiplicative interaction was suggested between GSTP1 and smoking on the risk of prostate cancer with the adjusted OR for the interaction of 4.52 (95% CI: 1.07 19.17). To our knowledge, this is the first time that a potential effect modification by the GSTP1 Ile/Ile genotype on smoking and the risk of prostate cancer is suggested. PMID- 15542264 TI - Prostate cancer mortality in Connecticut, Iowa and New Mexico African American men. AB - We sought to assess trends in prostate cancer incidence, treatment and mortality in African American men by means of analysis of prostate cancer data from three states, Connecticut, Iowa and New Mexico, all participants in the Surveillance, Epidemiology, and End Results (SEER) Program. Compared with levels before prostate specific antigen (PSA) testing, prostate cancer incidence increased in all three states after widespread testing. For men diagnosed with localized or regional prostate cancer, the respective increases in radical prostatectomy in Connecticut, Iowa, and New Mexico were 3.2, 2.3, and 4.9 times pre-test levels. Age-standardized mortality in Connecticut and Iowa increased slightly; in New Mexico the 104.7 deaths per 100,000 in 1979-1986, 62.1 in 1987-1990, dropped to 47.6 in 1991-1998, an amount of decline that was statistically significant. Introduction of PSA testing influenced early detection and treatment of prostate cancer in all three states. Although decline in prostate cancer mortality in New Mexico over time may be linked with use of the PSA test and definitive therapy, the relationship among these factors, and thus the proper treatment for the early stages of this condition, is unclear on the basis of these data. PMID- 15542266 TI - Neuropathological examination in forensic context. AB - Different diseases of and trauma to the central nervous system (CNS), as well as their consequences are common causes of death and therefore it is important to examine the CNS appropriately in forensic autopsy, bearing in mind that the site of the disease is often as crucial as its nature. The CNS is a complex organ and its examination requires special methods and knowledge and often consultation with a neuropathologist. The prerequisite for the proper examination is correct handling and processing of the CNS. Because of its soft consistency fixation of the CNS in toto before detailed macroscopic analysis is recommended but guidance for an expedited limited examination is also given. The key features to which attention should be paid during the removal and later macroscopic examination of the CNS are described. Processing CNS for microscopy also requires special techniques and in addition to the routine stains both special histological and selected immunohistochemical stainings are often needed to reach the correct diagnosis. PMID- 15542267 TI - Inflicted head injury in infants. AB - There is scant neuropathological information in the child abuse literature; even the best reviews include assumptions based on the findings of a few inadequate early studies. Our recent series of 53 fatal cases (Brain 124 (2001) 1290, 1299 [1,2]) demonstrated age-related patterns of brain injury and showed the substrate of severe encephalopathy in the infants to be hypoxic brain damage, not diffuse traumatic axonal injury ('DAI'), as had previously been thought. About one-third had craniocervical injuries, particularly in the brain stem, suggestive of stretch injury to the neuraxis. Our interpretation was that this finding implied a mechanism of injury--brain stem damage from stretch injury to the neck with resultant apnoea--that could account for the clinical scenario in many cases, and for which violence would not necessarily be required. Since publishing this study we have turned our attention to the subgroup of infants who die without objective signs of injury, such as skull fracture or impact, whose carers are accused of abuse, usually, "violent shaking", on the pathologic findings alone. Given the striking discrepancy that there often is in such cases between the relatively trivial findings in the brain and the accusations of violence, we have been looking at the pathogenesis of the typical intracranial bleeding. A histologic study of dura from 50 paediatric autopsies, none of whom had suffered a head injury, has led us to propose that the subdural and retinal bleeding in such cases may well have a physiological aetiology, rather than being caused directly by trauma. PMID- 15542268 TI - Trials and tribulations of using beta-amyloid precursor protein immunohistochemistry to evaluate traumatic brain injury in adults. AB - Axonal pathology is increasingly identified by beta-amyloid precursor protein (betaAPP) immunohistochemistry in the brains of patients who may or may not have a history of trauma. The presence of betaAPP-IR(+) has been variously interpreted as either that diffuse traumatic axonal injury (TAI) is indeed a universal finding in cases of fatal traumatic brain injury (TBI) or there are other causes of betaAPP-IR(+) axons which under certain circumstances may be sufficient to mimic TBI and therefore make the medico-legal interpretation of certain cases very difficult. To address some of the uncertainties we have undertaken a detailed analysis of the amount and distribution of betaAPP immunohistochemistry in 63 cases of fatal TBI, 17 cases of patients dying after cardiac arrest, 12 cases dying in association with status epilepticus, 3 cases of carbon monoxide (CO) poisoning, 13 cases of hypoglycaemia and in 60 controls. Three patterns of betaAPP-IR(+) were identified. First, diffuse multi-focal, second, corresponding to the outline of an infarct or haematoma, and thirdly a mixture of the two. The first pattern was seen in cases of the lesser grades of TAI, CO poisoning, and hypoglycaemia, the second pattern in cases in which there was evidence of raised intracranial pressure and the third in cases of severe TAI. It is concluded that the proper interpretation of cases requires the examination of a sufficient number of blocks ( [Formula: see text] ), processing using standardised protocols including betaAPP immunohistochemistry and in some cases the mapping of any IR(+) on anatomical line diagrams. betaAPP carried out on a small number of randomly taken blocks is likely to lead to misinterpretation of the clinico-pathological correlations and possibly to a miscarriage of justice. PMID- 15542269 TI - Long-term intracerebral inflammatory response after traumatic brain injury. AB - Epidemiological and pathological studies suggest that head injury is a significant risk factor for subsequent neurodegeneration and cognitive decline in later life. The precise mechanisms for the development of post-traumatic neurodegenerative change are unclear but we hypothesize that persistence of inflammatory processes in the brain may play a key role and that some individuals are more susceptible to such changes based on their genetic make-up. In support of this hypothesis we present evidence of persistent elevated microglial activity in long-term survivors of head injury and the suggestion of an association between the extent of this activity and interleukin-1 genotype. PMID- 15542270 TI - Hypoglycemic brain damage. AB - Hypoglycemia was long considered to kill neurons by depriving them of glucose. We now know that hypoglycemia kills neurons actively from without, rather than by starvation from within. Hypoglycemia only causes neuronal death when the EEG becomes flat. This usually occurs after glucose levels have fallen below 1 mM (18 mg/dl) for some period, depending on body glycogen reserves. At the time that abrupt brain energy failure occurs, the excitatory amino acid aspartate is massively released into the limited brain extracellular space and floods the excitatory amino acid receptors located on neuronal dendrites. Calcium fluxes occur and membrane breaks in the cell lead rapidly to neuronal necrosis. Significant neuronal necrosis occurs after 30 min of electrocerebral silence. Other neurochemical changes include energy depletion to roughly 25% of control, phospholipase and other enzyme activation, tissue alkalosis and a tendency for all cellular redox systems to shift towards oxidation. The neurochemistry of hypoglycemia thus differs markedly from ischemia. Hypoglycemia often differs from ischemia in its neuropathologic distribution, a phenomenon applicable in forensic practice. The border-zone distribution of global ischemia is not seen, necrosis of the dentate gyrus of the hippocampus can occur and a predilection for the superficial layers of the cortex is sometimes seen. Cerebellum and brainstem are universally spared in hypoglycemic brain damage. Hypoglycemia constitutes a unique metabolic brain insult. PMID- 15542271 TI - Gunshot injuries to the head and brain caused by low-velocity handguns and rifles. A review. AB - OBJECTIVES: Reconstruction of brain injuries is a basic task of forensic neuropathology. For better understanding of the wound ballistics of gunshot injuries to the brain caused by low-velocity firearms (E(o) < 550 J), we reviewed the respective contributions of: (1) biomechanical reconstruction by postmortem imaging techniques, (2) biometry of the extent of very early microscopic tissue destruction, and (3) microscopic studies on the type and extent of early microscopic reactions around the permanent missile track. MATERIAL AND METHODS: A selected case material of 47 victims of lethal gunshot wounding to the brain was studied. (1) Computed tomography (CT) and magnetic resonance imaging (MRI) techniques were compared with macroscopic findings in 17 cases. (2) Morphometric evaluation of the zones of cellular and axonal destruction around the permanent track was performed in 20 cases (survival time: <90 min). (3) Microscopic studies of the emigration of leukocytes and macrophages plus axonal expression of beta amyloid precursor protein (beta-APP) were conducted in 10 cases (survival time: >90 min). RESULTS AND CONCLUSIONS: (1) Imaging procedures provided valuable information on entrance and exit wounds, the missile track and secondary changes. (2) Biometry revealed a destruction zone of ca. 3.6 cm around the permanent track corresponding to the "temporary cavity". (3) Microscopic studies of reactive changes demonstrated axonal injury at sites remote from the permanent cavity that could explain the very early respiratory arrest following low-velocity gunshot injury. PMID- 15542272 TI - Accumulation of plasma proteins in Purkinje cells as an indicator of blood-brain barrier breakdown. PMID- 15542273 TI - Non-traumatic forensic neuropathology. AB - The practicing Forensic Pathologist is likely to encounter case material in which either the cause of death or a major contribution to the cause of death is underlying damage to or disease of the central nervous system. While it is good practice in many instances to have a working relationship with a Department of Neuropathology, from which advice and practical help can be sought, there may be instances when the Forensic Pathologist needs to proceed on a basis of a working knowledge of Forensic Neuropathology up to and including how to examine the specimen and take tissue blocks for processing and subsequent histological examination. Some of the more common conditions of the central nervous system such as damage consequent to hypoxia-ischaemia, hypoglycemia and epilepsy, the encephalopathies associated with altered sodium concentration, deficiency due to Vitamin B(1) and various neurodegenerative diseases that manifest as dementia and include Alzheimer's disease, cortical Lewy body disease and the prion disorders, are outlined in this article. PMID- 15542274 TI - The year in cardiac imaging. PMID- 15542275 TI - Leukocyte count and coronary heart disease: implications for risk assessment. AB - Inflammation is a key feature of atherosclerosis and its clinical manifestations. The leukocyte count is a marker of inflammation that is widely available in clinical practice. This paper reviews the available epidemiologic evidence for a relationship between the leukocyte count and coronary heart disease (CHD). Numerous epidemiologic and clinical studies have shown leukocytosis to be an independent predictor of future cardiovascular events, both in healthy individuals free of CHD at baseline and in patients with stable angina, unstable angina, or a history of myocardial infarction. This relationship has been observed in prospective and retrospective cohort studies, as well as in case control studies. It is strong, consistent, temporal, dose-dependent, and biologically plausible. The relationship persists after adjustment for multiple CHD risk factors, including smoking. Elevated differential cell counts, including eosinophil, neutrophil, and monocyte counts, also predict the future incidence of CHD. Leukocytosis affects CHD through multiple pathologic mechanisms that mediate inflammation, cause proteolytic and oxidative damage to the endothelial cells, plug the microvasculature, induce hypercoagulability, and promote infarct expansion. In summary, leukocytosis has been consistently shown to be an independent risk factor and prognostic indicator of future cardiovascular outcomes, regardless of disease status. The leukocyte count is inexpensive, reliable, easy to interpret, and ordered routinely in inpatient and outpatient settings. However, its diagnostic and prognostic utility in CHD is widely unappreciated. Further studies are needed to assess the true impact of leukocytosis on CHD, compare it with other inflammatory markers such as C reactive protein and lipoprotein phospholipase A(2) levels, and promote its use in CHD prediction. PMID- 15542276 TI - A time of accelerated change in academic cardiovascular medicine: implications for academic divisions of cardiology and their training programs. AB - With the vast epidemic of vascular disease predicted to be the leading cause of death and disability by a growing margin over the next 30 years, academic medical centers with cardiology training programs have a special responsibility. Given the dramatic advances of biotechnology in producing highly effective but expensive strategies of prevention and treatment, cardiovascular specialists should assist their academic centers in envisioning the future to prepare trainees for a different environment. Cardiologists of the future must be able to adapt to a societal need for patient-oriented, team-based clinical care and rapidly evolving technology, while maintaining the fundamental skills and knowledge required for individual patient interaction. Academic programs should benchmark their activities to ensure responsible resource allocation so that cardiologists of the future will be trained in an environment stimulating excellence and creativity. PMID- 15542277 TI - Effect of two different neuroprotection systems on microembolization during carotid artery stenting. AB - OBJECTIVES: This study sought to compare the efficacy of two different cerebral protection systems for the prevention of embolization during carotid artery stenting (CAS) using a transcranial Doppler (TCD) monitoring with the detection of microembolic signals (MES). BACKGROUND: Despite the introduction of cerebral protection systems, neurologic complications during CAS cannot completely be prevented. Transcranial Doppler and detection of MES may aid in assessing the efficacy of different neuroprotection systems. METHODS: A total of 42 patients with internal carotid artery stenoses were treated by CAS using either a filter (E.P.I. FilterWire, Boston Scientific Corp., Santa Clara, California) (n = 21) or a proximal endovascular clamping device (MO.MA system, Invatec s.r.l., Roncadelle, Italy) (n = 21). Microembolic signal counts were compared during five phases: placement of the protection device, passage of the stenosis, stent deployment, balloon dilation, and retrieval of the protection device. RESULTS: There were no significant differences in clinical or angiographic outcomes between the two groups. Compared to the filter device, the MO.MA system significantly reduced MES counts during the procedural phases of wire passage of the stenosis, stent deployment, balloon dilation, and in total (MES counts for the filter device were 25 +/- 22, 73 +/- 49, 70 +/- 31, and 196 +/- 84 during the three phases and in total, MES counts for the MO.MA system were 1.8 +/- 3.2, 11 +/- 19, 12 +/- 21, and 57 +/- 41, respectively; p < 0.0001). CONCLUSIONS: In comparison to a filter device the MO.MA system led to significantly lower MES counts during CAS. The detection of MES by TCD may facilitate the evaluation and comparison of different neuroprotection systems. PMID- 15542278 TI - Prognostic value of osteoprotegerin in heart failure after acute myocardial infarction. AB - OBJECTIVES: We sought to determine the relationship between osteoprotegerin (OPG) and clinical outcomes in patients with heart failure (HF) after acute myocardial infarction (AMI). BACKGROUND: Arterial calcification is a prominent feature of arterial atherosclerosis and is associated with the occurrence of AMI. Osteoprotegerin is a recently discovered member of the tumor necrosis superfamily that may link the skeletal with the vascular system. METHODS: We assayed plasma OPG levels in 234 patients with AMI complicated with HF and their relation to adverse outcomes during follow-up in patients randomly assigned to angiotensin converting enzyme inhibition or angiotensin II antagonism. Blood was sampled at baseline (median three days after AMI), one month, and at one and two years. RESULTS: Elevated plasma levels of OPG at baseline were associated with adverse outcomes during a median of 27 months follow-up; OPG remained an independent prognostic indicator also after adjustment for other known predictors of mortality and cardiovascular events after AMI (e.g., creatinine clearance, N terminal B-type natriuretic peptide, high-sensitivity C-reactive protein). In non survivors, plasma OPG levels were persistently elevated during longitudinal testing, suggesting that OPG may be of value for monitoring patients at risk. CONCLUSIONS: Osteoprotegerin is a novel marker for cardiovascular mortality and clinical events in patients with AMI complicated with HF. These findings are compatible with the hypothesis suggesting a possible association between mediators of bone homeostasis and cardiovascular disease. PMID- 15542279 TI - Skeleton key to vascular disease. PMID- 15542280 TI - Detection of exercise-induced ischemia by changes in B-type natriuretic peptides. AB - OBJECTIVES: The purpose of this study was to examine the effect of exercise induced ischemia on levels of B-type natriuretic peptide (BNP) and its inactive N terminal fragment (NT-pro-BNP)and to determine whether measurement of these peptides can improve the diagnostic accuracy of exercise testing. BACKGROUND: The ability of exercise testing to detect coronary artery disease (CAD) is limited by modest sensitivity and specificity. B-type natriuretic peptides (NT-pro-BNP and BNP) are released by ventricular myocytes in response to wall stress. We hypothesized that exercise-induced ischemia results in increased wall stress and triggers release of NT-pro-BNP and BNP. METHODS: A total of 74 patients with known CAD, normal left ventricular function, and normal resting levels of NT-pro BNP and BNP who were referred for exercise testing with radionuclide imaging, and 21 healthy volunteers, were enrolled. Blood was drawn before and after maximal exercise and analyzed for NT-pro-BNP and BNP. RESULTS: Of the patients with CAD, 40 had ischemia on perfusion images and 34 did not. Median post-exercise increases in NT-pro-BNP and BNP (DeltaNT-pro-BNP and DeltaBNP) were approximately four-fold higher in the ischemic group than in the nonischemic group (DeltaNT-pro BNP 14.5 vs. 4 pg/ml, p < 0.0001; DeltaBNP 36.5 vs. 7.5 pg/ml, p < 0.0001). In volunteers, median DeltaNT-pro-BNP was almost identical to that of the nonischemic patient group. At equal specificity to the electrocardiogram (ECG) (58.8%), the sensitivities of DeltaNT-pro-BNP and DeltaBNP for detecting ischemia were 90% and 80%, respectively; in contrast, the sensitivity of the exercise ECG was 37.5%. CONCLUSIONS: Measurement of exercise-induced increases in BNPs more than doubles the sensitivity of the exercise test for detecting ischemia with no loss of specificity. PMID- 15542281 TI - Acute changes in circulating natriuretic peptide levels in relation to myocardial ischemia. AB - OBJECTIVES: The aim of this study was to determine the effect of transient myocardial ischemia on circulating natriuretic peptide levels. BACKGROUND: Natriuretic peptides are released by the heart in response to wall stress. We hypothesized that transient myocardial ischemia would cause acute changes in circulating natriuretic peptide levels. METHODS: B-type natriuretic peptide (BNP), N-terminal fragment of BNP pro-hormone (NT-pro-BNP), and N-terminal fragment of atrial natriuretic peptide pro-hormone (NT-pro-ANP) levels were measured in 112 patients before, immediately after, and 4 h after exercise testing with nuclear perfusion imaging. RESULTS: Baseline levels of BNP were associated with the subsequent severity of provoked ischemia, with median levels of 43, 62, and 101 pg/ml in patients with none, mild-to-moderate, and severe inducible ischemia, respectively (p = 0.03). Immediately after exercise, the median increase in BNP was 14.2 pg/ml in patients with mild-to-moderate ischemia (p = 0.0005) and 23.7 pg/ml in those with severe ischemia (p = 0.017). In contrast, BNP levels only rose by 2.3 pg/ml in those who did not develop ischemia (p = 0.31). A similar relationship was seen between baseline NT-pro-BNP levels and inducible ischemia, but the changes in response to ischemia were less pronounced. NT-pro-ANP levels rose with exercise in both ischemic and non ischemic patients. When added to traditional clinical predictors of ischemia, a post-stress test BNP >or=80 pg/ml remained a strong and independent predictor of inducible myocardial ischemia (odds ratio 3.0, p = 0.025). CONCLUSIONS: Transient myocardial ischemia was associated with an immediate rise in circulating BNP levels, and the magnitude of rise was proportional to the severity of ischemia. These findings demonstrate an important link between the severity of an acute ischemic insult and the circulating levels of BNP. PMID- 15542282 TI - Serum levels of thiobarbituric acid reactive substances predict cardiovascular events in patients with stable coronary artery disease: a longitudinal analysis of the PREVENT study. AB - OBJECTIVES: The objective of this study was to test the predictive value of an oxidative stress biomarker in 634 patients from the Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial (PREVENT). BACKGROUND: Oxidative stress contributes to mechanisms of atherosclerosis and plaque instability. Biomarkers of oxidation, such as malondialdehyde (MDA), may represent independent indicators of risk for patients with stable coronary artery disease (CAD). METHODS: Serum MDA levels were measured as thiobarbituric acid reactive substances (TBARS) in 634 patients with documented CAD using reverse phase high-performance liquid chromatography and spectrophotometric approaches. RESULTS: During the three-year study, there were 51 major vascular events such as fatal/nonfatal myocardial infarction, 149 hospitalizations for nonfatal vascular events, and 139 patients underwent a major vascular procedure. At baseline, patients with TBARS levels in the highest quartile had a relative risk (RR) of 3.30 (95% confidence interval [CI] 1.47 to 7.42; p = 0.038) for major vascular events, RR of 4.10 (95% CI 2.55 to 6.60; p < 0.0001) for nonfatal vascular events, and RR of 3.84 (95% CI 2.56 to 5.76; p < 0.0001) for major vascular procedures. The effect of TBARS on events and procedures was also seen in a multivariate model adjusted for inflammatory markers (C-reactive protein, soluble intercellular adhesion molecule-1, interleukin-6), and other risk factors (age, low-density lipoprotein, high-density lipoprotein, total cholesterol, triglycerides, body mass index, and blood pressure). This analysis showed an independent effect of TBARS on major vascular events (p = 0.0149), nonfatal vascular events (p < 0.0001), major vascular procedures (p < 0.001), and all vascular events and procedures (p < 0.0001). CONCLUSIONS: Serum levels of TBARS were strongly predictive of cardiovascular events in patients with stable CAD, independently of traditional risk factors and inflammatory markers. PMID- 15542283 TI - The association between cardiorespiratory fitness and C-reactive protein in subjects with the metabolic syndrome. AB - OBJECTIVES: We sought to study relationship between cardiorespiratory fitness and C-reactive protein (CRP) in subjects with the metabolic syndrome. BACKGROUND: Recent studies have shown an association between the metabolic syndrome and chronic subclinical inflammation, as determined by elevated CRP. Cardiorespiratory fitness is associated with a lower risk of diabetes and improved insulin resistance. METHODS: Physical fitness was assessed in 1,640 subjects using the Bruce treadmill protocol and expressed as maximal metabolic equivalents. The level of CRP was measured using a high-sensitivity assay. RESULTS: Geometric mean CRP was calculated across quartiles of fitness after adjustment for age, gender, smoking, use of medications, and coronary disease. A strong inverse trend toward decreasing CRP levels with increasing fitness quartiles was present in subjects without metabolic abnormalities, subjects with one or two metabolic abnormalities, and subjects with the metabolic syndrome (all p 50%, n = 201), or those receiving angiotensin receptor antagonists (n = 51). Patients were genotyped for the ACE D/I polymorphism, followed to the end point of death or cardiac transplantation, and transplant-free survival compared by genotype. RESULTS: The ACE-D allele was associated with an increased risk of events (p = 0.026). In analysis by ACE inhibitor dose, this effect was primarily in the low-dose group (1-year percent event-free survival: II/ID/DD = 86/77/71,2-year = 79/66/59, p = 0.032). In the standard-dose group, the impact was markedly diminished (1-year: II/ID/DD = 91/81/80, 2-year: 77/70/71, p = 0.64). The impact of beta-blockers and high dose ACE inhibitors was greatest in subjects with the ACE DD genotype (p = 0.001) and was less apparent with the II and ID genotypes (p = 0.38). CONCLUSIONS: Higher doses of ACE inhibitors diminished the impact of the ACE-D allele, and the benefits of beta-blockers and high-dose ACE inhibitors appeared maximal for DD patients. Determination of ACE genotype may help target therapy for patients with heart failure. PMID- 15542287 TI - Regional coronary flow and contractile reserve in patients with idiopathic dilated cardiomyopathy. AB - OBJECTIVES: The purpose of this study was to assess regional coronary flow and contractile reserve in patients with idiopathic dilated cardiomyopathy (IDCM). BACKGROUND: Although IDCM has been associated with alterations in coronary blood flow and contractile reserve, little is known about their regional distribution and correlation. METHODS: Fourteen patients with IDCM and 11 control subjects underwent coronary flow velocity (APV) measurements in the left anterior descending (LAD), left circumflex (LCx), and right coronary (RCA) arteries at baseline (b) and at maximal hyperemia (h). Coronary flow reserve (CFR) was defined as h-APV/b-APV. Wall thickening was assessed in 16 segments (7 assigned to LAD, 5 to LCx, and 4 to RCA) both at rest and under peak stress during low dose dobutamine echocardiography. Regional contractile reserve was defined as the percentage difference in wall motion score index between rest and stress in each vascular territory. RESULTS: Although there were no significant differences in b APV, patients with IDCM had significantly lower h-APV than controls in all three vascular territories and reduced CFR (LAD: 2.79 +/- 0.43 vs. 3.48 +/- 0.51, p < 0.05; LCx: 2.71 +/- 0.39 vs. 3.36 +/- 0.65, p < 0.05; and RCA: 3.43 +/- 0.55 vs. 4.02 +/- 0.73, p < 0.05). There was also a significant correlation between CFR and the corresponding contractile reserve in the vascular territory of the LAD (r = 0.75, p = 0.002) and the LCx (r = 0.64, p = 0.014). CONCLUSIONS: Patients with IDCM have alterations in regional coronary flow and reduced CFR. Furthermore, the correlation between regional CFR and the corresponding contractile reserve indicates that microvascular dysfunction may have a pathophysiologic role in the evolution of the disease. PMID- 15542288 TI - Severe disease expression of cardiac troponin C and T mutations in patients with idiopathic dilated cardiomyopathy. AB - OBJECTIVES: We performed genetic investigations of cardiac troponin T (TNNT2) and troponin C (TNNC1) in 235 consecutive patients with idiopathic dilated cardiomyopathy (DCM) to evaluate prevalence of mutations and associated disease expression in affected families. BACKGROUND: Recently, mutations in sarcomeric genes have been reported in DCM. However, the prevalence, penetrance, and clinical significance of sarcomere gene mutations in large consecutive cohorts of DCM patients are poorly defined. METHODS: Mutation detection was performed by fluorescent SSCP/DHPLC analysis and direct sequencing. The functional effects of mutations on interactions within the troponin complex were assessed by a two hybrid luciferase assay. RESULTS: A total of 43% (102 of 235) of the study cohort had familial DCM. One TNNC1 and four TNNT2 (three novel) mutations were identified in one and four families, respectively. The prevalence of TNNC1/TNNT2 mutations in familial DCM was 5% with a penetrance of 100%. A total of 21 mutation carriers were identified; 6 underwent cardiac transplantation, 5 died of heart failure, and 4 died suddenly at a mean age of 29 years, while 6 remained stable on medication. Functional studies showed significant impairment of mutated troponin interaction compared with wild-type control, indicating an altered regulation of myocardial contractility. CONCLUSIONS: Cardiac troponin C was identified as a novel DCM gene. The disease expression associated with TNNC1 and TNNT2 mutations was severe with complete penetrance. The data suggest that mutation analysis of the troponin complex in DCM patients may prove valuable in early identification of individuals with an adverse prognosis and a high risk of premature death. This may lead to improved management and survival. PMID- 15542289 TI - Genetics of dilated cardiomyopathy: more genes that kill. PMID- 15542290 TI - The case for surgery in obstructive hypertrophic cardiomyopathy. AB - Relief of left ventricular (LV) outflow obstruction in patients with hypertrophic cardiomyopathy (HCM) and disabling symptoms refractory to maximum medical management has historically been a surgical problem. Surgical septal myectomy permanently abolishes systolic anterior motion of the mitral valve and mitral regurgitation, while normalizing LV pressures and wall stress. Also, these salutary goals are achieved without encumbering patients with post-procedural devices (e.g., pacemakers or defibrillators) or creating potentially arrhythmogenic substrates, as may occur with alcohol septal ablation. Procedural morbidity and mortality risk with myectomy is similar to, and in some institutions less than those for alcohol septal ablation. Over four decades, reports from numerous centers worldwide have consistently and unequivocably documented the benefits of surgery on hemodynamic and functional state, restoring normal and acceptable quality of life to patients of all ages by largely reversing the complications of heart failure. Long-term survival after myectomy is similar to that of the general population and superior to non-operated patients with obstruction. The LV outflow tract morphology in HCM is heterogeneous and not uncommonly includes congenital anomalies of the mitral valve apparatus for which the surgeon has the flexibility to adapt the repair, often employing an extended myectomy. In the current atmosphere of increasing and perhaps excessive enthusiasm for newer catheter-based interventions, it is a critical time to promote and re-emphasize that surgery is the time-honored (and presently the most effective) treatment strategy for relieving heart failure related disability resulting from dynamic LV outflow obstruction in HCM, and is the primary treatment option for this subgroup of severely symptomatic drug refractory patients. PMID- 15542291 TI - New treatment strategies for hypertrophic obstructive cardiomyopathy: alcohol ablation of the septum: the new gold standard? AB - Hypertrophic cardiomyopathy is a primary myocardial disorder with an autosomal pattern of inheritance characterized by inappropriate myocardial hypertrophy. Annual mortality has been reported to be 1% to 2% and sudden death represents the most common cause. Treatment strategies are 1) medical therapy in patients with mild to moderate symptoms, 2) reduction of septal hypertrophy by surgical myectomy or alcohol ablation, and 3) implantation of an automatic cardioverter defibrillator in the presence of non-sustained ventricular tachyarrhythmias. A debate has been started on whether surgical myectomy or alcohol ablation of the septum is the appropriate treatment for hypertrophic obstructive cardiomyopathy. Surgical (transaortic) myectomy has been the gold standard in the past 20 to 30 years for treatment of symptomatic patients with significant hemodynamic outflow tract obstruction. However, modern interventional technologies allow reduction of the myocardial septum by injection of alcohol into the first or second septal branch under guidance of two-dimensional (2D)-contrast echocardiography. This percutaneous technique not only has a lower morbidity than surgical myectomy but can be guided precisely by 2D echocardiography. One potential complication is transient (<30%) or permanent (<10%) atrioventricular block III; however, this complication is relatively rare. A randomized trial comparing the two treatment modalities is lacking, and the chance is small that such a trial will be performed because alcohol ablation can be done with high success and low complication rates, leaving only complex interventions (with valvular reconstructions and so on) for surgical myectomy. PMID- 15542292 TI - Global impairment of brachial, carotid, and aortic vascular function in young smokers: direct quantification by high-resolution magnetic resonance imaging. AB - OBJECTIVES: The purpose of this study was to assess vascular dysfunction in young smokers by high-resolution magnetic resonance imaging (MRI). BACKGROUND: Cigarette smoking is a well-known cause of endothelial dysfunction, reflected by impaired brachial artery reactivity to hyperemia. We hypothesized that smoking induces both peripheral endothelial dysfunction and altered function in central conduit arteries, and that these global changes in vascular function could be directly quantified in a single noninvasive examination using high-resolution MRI. METHODS: A total of 22 healthy young volunteers (mean age 31 +/- 2 years; 12 nonsmokers, 10 smokers: cumulative cigarette consumption 11.9 +/- 6.0 pack-years) underwent noninvasive high-resolution MRI to assess central vascular distensibility and pulse-wave velocity (PWV), and cross-sectional flow-mediated dilation (FMD) of the brachial artery. RESULTS: Brachial artery FMD was significantly reduced in smokers compared with nonsmokers (7.5 +/- 2.7% vs. 15.5 +/- 2.0%; p = 0.03), indicating impaired endothelium-dependant relaxation, whereas endothelium-independent responses to sublingual glyceroltrinitrate(400 mug) were identical in both groups. Impaired peripheral endothelial function in smokers was accompanied by striking decreases in central vascular distensibility in both the common carotid arteries (-45.7%; p = 0.02) and at multiple sites in the aorta (ascending aorta -26.9%, p = 0.04; thoracic descending aorta -25.0%, p = 0.01; abdominal descending aorta -25.5%, p = 0.02). Aortic arch PWV in smokers was increased by 19% (p = 0.02). CONCLUSIONS: Cigarette smoking induces global changes in both peripheral and central vascular function. Direct quantification of multiple parameters of vascular function using high-resolution MRI will provide powerful new approaches to the assessment of vascular disease pathogenesis, diagnosis, and treatment. PMID- 15542293 TI - Cardiac transplantation after the Fontan or Glenn procedure. AB - OBJECTIVES: The purpose of this study was to review the clinical course and outcome of cardiac transplantation after a failed Glenn or Fontan procedure. BACKGROUND: Late complications of the Glenn or Fontan procedure, including ventricular failure, cyanosis, protein-losing enteropathy, thromboembolism, and dysrhythmias often lead to significant morbidity and mortality. If other therapies are ineffective, cardiac transplantation is the only therapeutic recourse. Transplantation in this unique population presents significant challenges in the operative and perioperative periods. METHODS: The anatomic diagnoses, previous operations, clinical status, and indications for transplantation were characterized in patients transplanted after a Glenn or Fontan procedure. Outcomes after transplantation, including postoperative complications and mortality, were reviewed. Comparisons were made between survivors and nonsurvivors. RESULTS: Primary orthotopic cardiac transplantation was performed in 35 patients (mean age 15.7 +/- 8.5 years) with a mean follow-up of 54 +/- 46 months. A total of 11 patients had undergone a Glenn shunt and 24 patients a Fontan procedure. Indications for transplantation were a combination of causes including ventricular dysfunction, failed Fontan physiology, and/or cyanosis. Ten patients died 3.5% (w/v), corresponding to opaque gel above 30 degrees C, a percolated structure, i.e., network-like domain is formed by NE as a result of macrophase separation due to dehydration of the PNIPA chains. As the temperature increases toward 40 degrees C, the network domain is squeezed along a direction parallel to the NE interface, which leads to increase of the interfacial thickness given by swollen PEG chains and to the macroscopic synerisis behavior. PMID- 15542329 TI - Re-entrant volume phase transition of hydrogel membrane of microcapsule. AB - Re-entrant volume phase transition of hydrogel wall membrane of microcapsules (MC) was first observed using MC suspensions consisting of poly (L-lysine-alt terephthalic acid) wall and aqueous inner and outer solutions with different pHs. To analyze the dynamics of the re-entrant phase transition, we extended the theory for the swelling and the shrinking dynamics of the microcapsule gel [T. Narita, T. Yamamoto, D. Suzuki, T. Dobashi, Langmuir 19 (2004) 4051]. In the theory, the microcapsule size and the force constant for the driving force which gives rise to the size relaxation were chosen as the thermodynamic variables. The time course of the cross-sectional area of the microcapsules fitted well to the theoretical equations, and the time constants determined as the fitting parameters were discussed in terms of the force constant relaxation and the size relaxation. PMID- 15542330 TI - Metal ion absorption of carboxymethylcellulose gel formed by gamma-ray irradiation. For the environmental purification. AB - Carboxymethylcellulose (CMC) is well known as a safe and biodegradable material, which is widely used as food additives, wash paste, etc., in our daily life. Aiming at the environmental purification of this eco-friendly substance, the authors have investigated Cu2+ absorption property of CMC gel that was crosslinked by irradiating with gamma-rays without toxic crosslinker. The CMC gel has revealed to capture Cu2+ of which number depends on the gamma-ray dose as well as the CMC concentration, indicating the chelation by carboxyl group at the end of the side chain. PMID- 15542331 TI - The effect of the linear charge density of carrageenan on the ion binding investigated by differential scanning calorimetry, dc conductivity, and kHz dielectric relaxation. AB - The effect of the linear charge density of natural polyelectrolyte, carrageenan, on the ion binding to carrageenan molecules in relation to the gelation was investigated by using the dielectric relaxation spectroscopy, dc conductivity, optical rotation, and differential scanning calorimetry (DSC). Although carrageenan is an anionic polysaccharide, carrageenan molecules in the helix state at low temperatures can bind not only cation, such as potassium and cesium, but also anion, such as iodide. The dc conductivity steeply decreases just below the coil-helix transition temperature, which indicates the binding of ion to the carrageenan molecules in the helix state due to the increase of the linear charge density compared with that in the coil state. The addition of NaI promotes the helix formation, and prevents from aggregation of helices, which was suggested by the results of the dynamic shear modulus and the DSC, and resulted in an increase of the relaxation amplitude of the lowest frequency relaxation ( approximately kHz) attributed to the fluctuation of the tightly bound counter ions along the high charge density region (helix). It is concluded that binding of iodide induces (1) the increase in the amount of tightly bound counterions to carrageenan molecules and (2) the formation of non-aggregated helix. PMID- 15542332 TI - Effect of deacetylation rate on gelation kinetics of konjac glucomannan. AB - Effect of deacetylation rate on the gelation behaviors on addition of sodium carbonate for native and acetylated konjac glucomannan (KGM) samples with a degree of acetylation (DA) range of 1.38-10.1 wt.% synthesized using acetic anhydride in the presence of pyridine as catalyst was studied by dynamic viscoelastic measurements. At a fixed alkaline concentration (C(Na)), both the critical gelation times (t(cr)) and the plateau values of storage moduli (G'(sat)) of the KGM gels increased with increasing DA. While at a fixed ratio of alkaline concentrations to values of DA (C(Na)/DA), the similar t(cr) and (G'(sat)) values independent of DA were observed. On the whole, increasing KGM concentration or temperature shortened the gelation time and enhanced the elastic modulus for KGM gel. The effect of deacetylation rate related to the C(Na)/DA on the gelation kinetics of the KGM samples were discussed. PMID- 15542333 TI - Wetting characteristics of aqueous rhamnolipids solutions. AB - The wetting properties of surfactants on solid surfaces form the basis of many industrial and biological processes. The preferential adsorption of the surfactants from aqueous solutions onto solid surfaces alter the adhesion tension of the surface and this behavior may cause partial to complete wetting of the surfaces by the aqueous surfactant solutions. However, different types of surfactants show different wetting characteristics. To study the wetting properties of biologically produced rhamnolipids (RL), advancing contact angles of the aqueous solutions of the RL mixture of R1 and R2 in a ratio of R2/R1=1.1 were measured as a function of surfactant concentration. For a comparison of the wetting performance, sodium dodecyl sulfate (SDS) was chosen as the reference surfactant. A hydrophilic glass surface, a hydrophobic polymer, polyethylene terephthalate (PET), and gold surface were used as the solid surfaces to determine the wetting characteristics of rhamnolipids. At low surfactant concentrations (RL concentration <3x10(-5)M, SDS concentration<3x10(-4)M) contact angle (Theta) varied in a certain range depending on the character of the surfactant interactions with the surface. This was followed by a decrease in contact angle. Parallel to this behavior, at low surfactant concentrations the adhesion tension decreased, then remained constant and an increase at higher surfactant concentrations was obtained on hydrophobic surfaces. On hydrophilic surfaces a steady decrease in adhesion tension was observed with both surfactant solutions. PMID- 15542334 TI - Protein resistant polyurethane surfaces by chemical grafting of PEO: amino terminated PEO as grafting reagent. AB - The objective of this work was to gain a better understanding of the mechanism of resistance to protein adsorption of surfaces grafted with poly(ethylene oxide) (PEO). A polyurethane-urea was used as a substrate to which PEO was grafted. Grafting was carried out by introducing isocyanate groups into the surface followed by reaction with amino-terminated PEO. Surfaces grafted with PEO of various chain lengths (PUU-NPEO) were prepared and characterized by water contact angle and X-ray photoelectron spectroscopy (XPS). XPS data indicated higher graft densities on the PUU-NPEO surfaces than on analogous surfaces prepared using hydroxy-PEO (PUU-OPEO) as reported previously [J.G. Archambault, J.L. Brash, Colloids Surf. B: Biointerf. 33 (2004) 111-120]. Protein adsorption experiments using radiolabeled myoglobin, concanavalin A, albumin, fibrinogen and ferritin as single proteins in buffer showed that adsorption was reduced on the PEO-grafted surfaces by up to 95% compared to the control. Adsorption decreased with increasing PEO chain length and reached a minimum at a PEO MW of 2000. Adsorption levels on surfaces with 5000 and 2000 MW grafts were similar. There was no clear effect of protein size on resistance to protein adsorption. Adsorption on the PUU NPEO surfaces was significantly lower than on the corresponding PUU-OPEO surfaces, again suggesting higher graft densities on the former. Adsorption of fibrinogen from plasma was also greatly reduced on the grafted surfaces. From analysis (SDS-PAGE, immunoblotting) of the proteins eluted after plasma exposure, it was found that the grafted surfaces and the unmodified substrate adsorbed the same proteins in roughly the same proportions, suggesting that adsorption to the PEO surfaces occurs on patches of bare substrate. The PEO grafts did not apparently cause differential access to the substrate based on protein size. PMID- 15542335 TI - Preparation and properties of monodispersed rifampicin-loaded poly(lactide-co glycolide) microspheres. AB - Monodispersed rifampicin (RFP)-loaded poly(lactide-co-glycolide) (PLGA) microspheres were prepared by a solvent evaporation method. In order to control the sizes of the microspheres, a membrane emulsification technique using Shirasu porous glass (SPG) membranes was applied. RFP/PLGA microspheres with the average diameters of 1.3, 2.2, 5.2, and 9.0 microm were obtained. They were relatively monodisperse and the values of the coefficient of variation (CV) for the size distributions of the microspheres were in the range between 7.0 and 16.0%. The loading efficiency of RFP was in the range between 50.3 and 67.4% independent of the microsphere size. The release ratio of RFP from RFP/PLGA microspheres was measured in pH 7.4 PBS at 37 degrees C. From RFP/PLGA microspheres with average diameters of 1.3 and 2.2 microm, almost 60% of RFP loaded in the microspheres was released in the initial day and the release was terminated almost within 10 days. On the other hand, from those with average diameters of 5.2, and 9.0 microm, the release of RFP was observed even 20 days after the release started. PMID- 15542336 TI - Inhibition of heat-induced aggregation of a beta-lactoglobulin-stabilized emulsion by very small additions of casein. AB - Heat stability has been studied in model systems of oil-in-water emulsions (3 wt.% total protein, 45 vol.% n-tetradecane, pH 6.8, ionic strength 30-50 mM) with pure beta-lactoglobulin (beta-lg) as the main emulsifier. The effect of small additions of sodium caseinate, beta-casein or alpha s1-casein prior to emulsion preparation has been investigated. Samples heated for 3 min at 90 degrees C were monitored with respect to changes in viscosity and particle-size distribution. As expected, the pure beta-lg-stabilized emulsions were susceptible to heat-induced changes. But the replacement of just 1% of the beta-lg by sodium caseinate (0.03 wt.% caseinate in the total emulsion) led to complete elimination of any heat induced viscosity or particle size increase. These findings show that a very small proportion of casein can inhibit the susceptibility of a beta-lg-based emulsion to heat-induced destabilization. The magnitude of the effect is dependent on the type of casein, with the order of effectiveness being beta casein>sodium caseinate>alpha s1-casein. This work has potential implications for the development of milk protein-stabilized emulsions of improved shelf life. PMID- 15542337 TI - Preparation and characterization of metal-chitosan nanocomposites. AB - Various metal-chitosan nanocomposites were synthesized, including silver (Ag), gold (Au), platinum (Pt), and palladium (Pd) in aqueous solutions. Metal nanoparticles were formed by reduction of corresponding metal salts with NaBH4 in the presence of chitosan. And chitosan molecules adsorbing onto the surface of as prepared metal nanoparticles formed the corresponding metal-chitosan nanocomposites. Transmission electron microscopy (TEM) images and UV-vis spectra of the nanocomposites revealed the presence of metal nanoparticles. Comparison of all the resulting particles size, it shows that silver nanoparticles are much larger than others (Au, Pt and Pd). In addition, the difference in particles size leads to develop different morphologies in the films cast from prepared metal chitosan nanocomposites. Polarized optical microscopy (POM) images show a batonet like structure for Ag-chitosan nanocomposites film, while for the films cast from other metal (Au, Pt, and Pd)-chitosan nanocomposites, some branched-like structures with a few differences among them were observed under POM observation. PMID- 15542338 TI - Extractable substances from human hair: a discussion about the origin of the holes. AB - The existence of holes in the endocuticle, in the inter-macrofibrilar matrix and in the cell membrane complex of hair fibers is described. In this paper, we show that these holes are absent in the hair follicle. However, cell remnants found in young hair inside the follicle are located in the same regions and are of the same dimensions as the holes observed in mature fibers. Since holes in the endocuticle have been described to appear as a result of daily care actions, it is probable that the development of holes in the inter-macrofibrilar matrix and in the cell membrane complex is related to the removal of extractable substances. We discuss hole formation in terms of the nature of the extractable substances and the presence of cell remnants in the young fibers. PMID- 15542339 TI - E. coli adhesion to silica in the presence of humic acid. AB - The influence of humic acid on the adhesion of Escherichia coli to silica particles or glass surfaces was investigated. After adsorbing various amounts of humic acid to the particles or surfaces, bacteria were added to the sample and allowed to adhere. For the silica particles the number of bacteria-particle couplets formed were counted from video microscopy images. For the glass surfaces, a differential electrophoresis force was applied, and the force required to detach the bacteria was quantified. These experiments showed a slight increase in the number of couplets formed in the presence of humic acid, and also showed a slight increase in the force required for detachment of the bacteria. Although an increase in adhesion number and strength was measured, the magnitude of the increase was small, indicating that humic acid plays a small role in bacterial adhesion to silica or glass surfaces. PMID- 15542340 TI - A theory of overshoot phenomena in an ion-exchange membrane system. AB - A simple theory based on a kinetics equation with time delay is presented for ion permeation through an ion-exchange membrane for the case where two or more species of ions of the same charge are permeating from solutions 1 to 2 through the membrane. This theory explains the observation of Neihof and Sollner that if the exchange rates of ionic species are quite different from each other, then the concentration of the fastest exchanging species of ions in solution 2 exhibits overshoot [R. Neihof, K. Sollner, J. Phys. Chem. 61 (1957) 159]. PMID- 15542341 TI - Properties of various phosphatidylcholines as emulsifiers or dispersing agents in microparticle preparations for drug carriers. AB - We investigated characteristics of various phosphatidylcholines (PCs) used as dispersing agents and emulsifiers. Six PCs with different lengths of acyl hydrocarbon chains and different degrees of unsaturated acyl hydrocarbon chains were selected to examine influences of a lipophillic part of phosphatidylcholines in emulsion and dispersion systems. Vesicles and oil-in-water emulsions were prepared by sonication under several ambient temperature conditions. Mean diameters of vesicles and oil droplets in emulsions were measured by a submicron particle sizer. In vesicles that are generated by hydration of the PCs and have a bilayer form, particle size was influenced by length and degree of unsaturation of acyl hydrocarbon chains of a PC. PCs with shorter acyl hydrocarbon chains or unsaturated bonds are considered more potent dispersing agents. Preparation temperature of the PC is also a factor affecting potency of dispersion. In O/W emulsions in which PCs were absorbed at water-oil interfaces and which have a single layer form or liquid-crystal layer form, particle size was also influenced by length and degree of unsaturation of acyl hydrocarbon chains of a PC. PCs with shorter and saturated acyl hydrocarbon chains are considered more potent emulsifiers. Unsaturation of acyl hydrocarbon chains weaken the ability of emulsification due to vulnerable double bonds. For stable emulsions, it is considered beneficial for PCs to form small oil droplets and lamellae liquid crystal phase. From this perspective, saturated PCs with short hydrocarbon chains, i.e., DLPC and DMPC, may have advantages in preparing a stable emulsion not only by giving a smaller droplet size but also by forming lamellae liquid crystal phase. When considering characteristics of PCs as emulsifiers, their characteristics as dispersing agents is also useful information. PMID- 15542342 TI - Overshoot and oscillation in surface tension of gelatin solutions. AB - The experimental data of the surface tension of aqueous gelatin solution reported by Sato and Ueberreiter [H. Sato, K. Ueberreiter, Macromol. Chem. 180 (1979) 829; H. Sato, K. Ueberreiter, Macromol. Chem. 180 (1979) 1107; H. Sato, K. Ueberreiter, ACS Polym. Preprints 20 (1979) 907] were analyzed with a theory of adsorption kinetics with time delay. It is found that the observed overshoot and oscillation of the surface tension are well explained by the present theory. The delay time and the rate constants of adsorption and desorption of gelatin molecules onto the air/gelatin solution interface have been evaluated. PMID- 15542343 TI - Self-assembly and characterization of paclitaxel-loaded N-octyl-O-sulfate chitosan micellar system. AB - N-octyl-O-sulfate chitosan micellar system loaded paclitaxel was prepared by using dialysis method. The critical micelle concentration (CMC) of the modified chitosan was found to be 0.45 mg/ml. Compared with the amount of N-octyl-O sulfate chitosan, the paclitaxel loading amount in the system was up to 25% (w/w), depending on both of the solvents used in dialysis and the feed weight ratio of paclitaxel to the derivative. The polymeric micelles forming and loading occurred simultaneously in the dialysis process when ethanol and water were utilized as the solvents for paclitaxel and the polymer, respectively. Paclitaxel loaded micellar system of N-octyl-O-sulfate chitosan was characterized by DSC, WXRD and TEM. TEM photograph revealed that paclitaxel existed as the colloid particulates in ethanol before loading and in the cores of the spherical polymeric micelles after loading. The results of DSC and WXRD indicated that paclitaxel was transferred from the crystalline state to amorphous state after loading. The lyophilized powder of micellar system (25% (w/w) loading) could be reconstituted easily in aqueous media even after 2 months storage at 4 degrees C without the change of paclitaxel entrapment and micelle size. The reconstituted solution (2.1 mg paclitaxel/ml) also showed good stability. The dilution with saline may decrease the loading and physical stability based on the dilution times which was related with CMC of the polymer. In vitro tests showed that paclitaxel was slowly released from micellar solution and the release lasted up to 220 h by means of the dialysis method. PMID- 15542344 TI - Simulations of temperature dependence of the formation of a supported lipid bilayer via vesicle adsorption. AB - Recent experimental investigations of the kinetics of vesicle adsorption in solution on SiO2 demonstrate a thermally activated transition from adsorbed intact vesicles to a supported lipid bilayer. Our Monte Carlo simulations clarify the mechanism of this process. The model employed is an extension of the model used earlier to describe vesicle adsorption at room temperature. Specifically, it includes limitations of the adsorption rate by vesicle diffusion in the solution, and adsorption- and lipid-membrane-induced rupture of arriving and already adsorbed vesicles. Vesicles and lipid molecules, formed after rupture of vesicles, are considered immobile. With these ingredients, the model is able to quantitatively reproduce the temperature-dependent adsorption kinetics, including a higher critical surface concentration of intact vesicles for lower temperatures, and the apparent activation energy for the vesicle-to-bilayer transition E(a) approximately 5 kcal/mol. PMID- 15542345 TI - Chemical graft polymerization of sulfobetaine monomer on polyurethane surface for reduction in platelet adhesion. AB - Surface modification is an effective way to improve the hemocompatibility and remain bulk properties of biomaterials. Recently, polymer tailored with zwitterions was found having good blood compatibility. In this study, the zwitterionic monomer of sulfobetaine was graft polymerized onto polyurethane (PU) surface in a three-step heterogenous system through the vinyl bonds of acrylic acid (AA) or hydroxyethyl methacrylate (HEMA), which was immobilized with hexamethylene diisocyanate (HDI) beforehand. First, PU was activated with isocyanate groups using HDI as coupling agent. Second, AA or HEMA was introduced through reaction of AA or HEMA with NCO groups bonded on PU surface. Last, zwitterionic monomer of sulfobetain was graft polymerized with vinyl group of AA or HEMA using AIBN as polymerization initiator. The reaction process was monitored with ATR-IR spectra and XPS spectra. Variation of graft yield with temperature and monomer feed concentration was investigated and feasible conditions were optimized. The wettability of films was investigated by water contact angle measurement and water absorbance. Platelet adhesion experiment was conducted as a preliminary test to confirm the improved blood compatibility of PU. The number of platelets adhering to PU decreased greatly comparing with the originals after 1 and 3 h of contact with human plate-rich plasma (PRP). PMID- 15542346 TI - The influence of cetyltrimethyl ammonium bromide on electrochemical properties of thyroxine reduction at carbon nanotubes modified electrode. AB - The effect of cetyltrimethyl ammonium bromide (CTAB) on the electrochemical behaviors of thyroxine at a glassy carbon electrode (GCE) modified with single walled carbon nanotubes (SWNTs) was investigated. At the SWNTs film-coated GCE, a well-defined oxidation peak of thyroxine at 0.78 V was obtained, but the reduction peak of thyroxine was indiscernible. When trace CTAB was added to the working solution, the reduction current could be greatly enhanced and the oxidation current remained stable. The reaction mechanisms for the reduction of thyroxine were explored by chronocoulometry. Thyroxine might form particular ion complex with CTAB via the interaction between iodine atoms on thyroxine and bromide ions in CTAB, which made the concentration of thyroxine at the surface of the modified electrode increased and the electron transfer rate enhanced. The proper mechanisms for the enhanced reduction of thyroxine in the present of CTAB were explored by several electrochemical techniques including cycle voltammetry linear sweep voltammetry and others. It was concluded that the special interactions between the thyroxine CTAB and SWNTs resulted in the increase of the reduction peak current. All results indicated that two iodine atoms on the thyroxine and four electrons were involved the reduction process which was irreversible and two iodine ions produced. In this system, the sensitive reduction peak of thyroxine at 0.3 V was employed to determine thyroxine and a low detection limit of 2x10(-8) mol/L was obtained for 2 min accumulation at 0.9 V. The SWNTs coated GCE had good stability and reproducibility. PMID- 15542347 TI - Zinc deficiency, DNA damage and cancer risk. AB - A large body of evidence suggests that a significant percentage of deaths resulting from cancer in the United States could be avoided through greater attention to proper and adequate nutrition. Although many dietary compounds have been suggested to contribute to the prevention of cancer, there is strong evidence to support the fact that zinc, a key constituent or cofactor of over 300 mammalian proteins, may be of particular importance in host defense against the initiation and progression of cancer. Remarkably, 10% of the U.S. population consumes less than half the recommended dietary allowance for zinc and are at increased risk for zinc deficiency. Zinc is known to be an essential component of DNA-binding proteins with zinc fingers, as well as copper/zinc superoxide dismutase and several proteins involved in DNA repair. Thus, zinc plays an important role in transcription factor function, antioxidant defense and DNA repair. Dietary deficiencies in zinc can contribute to single- and double-strand DNA breaks and oxidative modifications to DNA that increase risk for cancer development. This review will focus on potential mechanisms by which zinc deficiency impairs host protective mechanisms designed to protect against DNA damage, enhances susceptibility to DNA-damaging agents and ultimately increases risk for cancer. PMID- 15542348 TI - Correlation of apolipoprotein M with leptin and cholesterol in normal and obese subjects. AB - Apolipoprotein M (apoM) is a recently characterized apolipoprotein that is exclusively expressed in the liver and kidney. In plasma it is present predominantly in high-density lipoprotein (HDL). The physiological function of apoM is not yet known. In the present study we investigated relationships between plasma apoM levels and leptin levels, body mass index (BMI), as well as fasting glucose and other lipid parameters in women with a wide range of BMI (18.9-57.1 kg/m(2), n = 51). In univariate analysis, apoM correlated significantly with leptin (r = 0.54, P < 0.001), BMI (r = 0,70, P < 0.001), fasting insulin (r = 0.33, P = 0.025), total cholesterol (r = -0.41, P = 0.016), and LDL-cholesterol (r = -0.39, P = 0.018). The correlations between apoM and cholesterol and between apoM and leptin remained significant after adjustment for the influence of BMI. Forward stepwise multiple regressions when leptin, BMI, insulin, and cholesterol were entered into a model as independent variables and apoM as the dependent variable, showed that cholesterol and leptin were independent predictors of circulating apoM. These two parameters yielded a value of r(2) = 0.28, thereby explaining approximately 30% of the variance in apoM. Hence, we show that apoM is positively related to leptin and negatively related to cholesterol in humans. PMID- 15542349 TI - Effects of soybean isoflavones, probiotics, and their interactions on lipid metabolism and endocrine system in an animal model of obesity and diabetes. AB - The effects of soybean isoflavones with or without probiotics on tissue fat deposition, plasma cholesterol, and steroid and thyroid hormones were studied in SHR/N-cp rats, an animal model of obesity, and were compared to lean phenotype. We tested the hypothesis that probiotics by promoting the conversion of isoflavone glycosides to their metabolically active aglycone form will have a synergistic effect on body fat, cholesterol metabolism, and the endocrine system. Obese and lean SHR/N-cp rats were fed AIN-93 diets containing 0.1% soy isoflavone mixture, 0.1% probiotic mixture, or both together. Different fat tissues were teased and weighed. Plasma was analyzed for cholesterol and steroid and thyroid hormones. In both phenotypes, isoflavones lowered fat deposition in several fat depots. Probiotics alone had no significant effect on fat depots. Isoflavones lowered total, LDL, and HDL cholesterol in lean rats, but in obese rats isoflavones lowered only total and LDL cholesterol. Isoflavones also lowered many of the steroid hormones involved in lipid metabolism but had no significant effect on thyroid hormones. Probiotics had no significant effect on cholesterol or hormones. Thus, our data show that soy isoflavones also lower plasma cholesterol and that this hypocholesterolemic effect appears to be due in part to the modulation of steroid hormones. Probiotics do not seem to enhance the effect of isoflavones. PMID- 15542350 TI - Biotin deficiency decreases life span and fertility but increases stress resistance in Drosophila melanogaster. AB - Biotin deficiency is associated with fetal malformations and activation of cell survival pathways in mammals. In this study we determined whether biotin status affects life span, stress resistance, and fertility in the fruit fly Drosophila melanogaster. Male and female flies of the Canton-S strain had free access to diets containing 6.0 (control), 4.8, 2.5, or 0 pmol biotin/100 mg. Biotin concentrations in diets correlated with activities of biotin-dependent propionyl CoA carboxylase and biotin concentrations in fly homogenates, but not with biotinylation of histones (DNA-binding proteins). Propionyl-CoA carboxylase activities and biotin concentrations were lower in males than in females fed diets low in biotin. The life span of biotin-deficient males and females was up to 30% shorter compared to biotin-sufficient controls. Exposure to oxidative stress reversed the effects of biotin status on survival in male flies: survival times increased by 40% in biotin-deficient males compared to biotin-sufficient controls. Biotin status did not affect survival of females exposed to oxidative stress. Exposure of flies to cold, heat, and oxidative stress was associated with mobilization of biotin from yet unknown sources. Biotin deficiency decreased fertility of flies. When biotin-deficient males and females were mated, the hatching rate (larvae hatched per egg) decreased by about 28% compared to biotin sufficient controls. These findings are consistent with the hypothesis that biotin affects life span, stress resistance, and fertility in fruit flies. PMID- 15542351 TI - Pulsed electric fields-processed orange juice consumption increases plasma vitamin C and decreases F2-isoprostanes in healthy humans. AB - Orange juice, a rich source of vitamin C, accounts for 60% of all fruit juices and juice-based drinks consumed in western Europe. Orange juice preservation is currently accomplished by traditional pasteurization. Pulsed electric fields (PEF) have been studied as a nonthermal food preservation method. Food technology needs in the area of processing are driven by nutrition. Therefore, the objectives of this study were to assess the bioavailability of vitamin C from pulsed electric fields-treated orange juice in comparison with freshly squeezed orange juice and its impact on 8-epiPGF(2alpha) concentrations (biomarker of lipid peroxidation) in a healthy human population. Six subjects consumed 500 mL/day of pulsed electric fields-treated orange juice and six subjects consumed 500 mL/day of freshly squeezed orange juice for 14 days, corresponding to an intake of about 185 mg/day of ascorbic acid. On the first day of the study, subjects drank the juice in one dose, and on days 2-14 they consumed 250 mL in the morning and 250 mL in the afternoon. Blood was collected every hour for 6 hours on the first day and again on days 7 and 14. In the dose-response study, the maximum increase in plasma vitamin C occurred 4 hours postdose. Vitamin C remained significantly higher on days 7 and 14 in both orange juice groups. Plasma 8-epiPGF(2alpha) concentrations was lower at the end of the study (P < 0.001) in both groups. Plasma levels of vitamin C and 8-epiPGF(2alpha) were inversely correlated. Pulsed electric fields-preservation of orange juice retains the vitamin C bioavailability and antioxidant properties of fresh juice with a longer shelf-life. PMID- 15542352 TI - Antioxidant role of zinc in PCB (Aroclor 1254) exposed ventral prostate of albino rats. AB - The ability of zinc to retard oxidative processes has been recognized for many years. Polychlorinated biphenyls (PCBs) are persistent and bioaccumulative environmental toxicants. Previous study has indicated that PCBs can have deleterious effects, including oxidative stress, on various aspects of reproduction in male rats. The aim of this study was to determine the antioxidant role of zinc in PCB-exposed ventral prostate of albino rats. A group of 20 rats were treated with Aroclor 1254 (2 mg/kg body weight/day, i.p.) for 30 days. After the PCB treatment, 10 rats were treated as PCB control. The remaining 10 rats were given zinc (Zn SO(4)) (200 mg/kg body weight/day, p.o.) for 10 days. Ventral prostatic enzymatic antioxidants such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) were estimated in all the groups. Hydrogen peroxide (H(2)O(2)), lipid peroxidation (LPO) and ventral prostatic acid phosphatase (ACP) were also estimated. Serum hormonal profiles such as total tri-iodothyronine (T(3)), thyroxine (T(4)), thyroid stimulating hormone (TSH), testosterone, and estradiol were estimated. Ventral prostatic androgen and estrogen receptors, ventral prostatic zinc content, and serum zinc concentration were also quantified in all the groups. Antioxidant enzymes such as SOD, CAT, GPx, GST, and ACP were decreased while an increase in H(2)O(2) and LPO were observed in PCB-treated animals. Decreased serum total T(3), T(4), testosterone, estradiol and increased TSH were observed in PCB-exposed rats. Ventral prostatic androgen and estrogen receptors were also decreased significantly in PCB-exposed rats. Zinc administration restored to previous levels all parameters except ventral prostatic ACP. These results suggest that PCB induces oxidative stress in rat ventral prostate by decreasing the levels of antioxidant enzymes; the effects could be reversed by the administration of zinc. The adverse effect of PCBs (Aroclor 1254) and zinc on ventral prostate might be due to indirect action through hormonal regulation. PMID- 15542353 TI - Characterization of diabetes-related traits in MSM and JF1 mice on high-fat diet. AB - We examined the effect of a high-fat diet on the diabetes-related traits of the Japanese Fancy mouse 1 (JF1), MSM, and C57BL/6J (B6J) mice. MSM and JF1 mice were derived from Mus musculus molossinus. B6J is a commonly used laboratory strain, with the vast majority of genome segments derived from Mus musculus domesticus and Mus musculus musculus, and is susceptible to high-fat diet-induced type 2 diabetes. None of the strains showed symptoms of diabetes or obesity when fed a laboratory chow diet. Under a high-fat diet, JF1 mice developed impaired glucose tolerance, hyperglycemia, hyperinsulinemia, and obesity. B6J mice fed a high-fat diet mildly developed these diabetes-related traits compared to JF1 mice fed a high-fat diet. JF1 mice fed a high-fat diet were classified as having type 2 diabetes and were susceptible to high-fat diet-induced diabetes and obesity. On the other hand, MSM mice were resistant to high-fat diet-induced diabetes. These results indicate that the JF1 strain, with its unique genetic origin, is a useful new animal model of high-fat diet-induced diabetes and obesity. Further investigations using JF1 mice will help to clarify the role of the high-fat diet on human diabetes and obesity. PMID- 15542354 TI - Oat (Avena sativa L.) and amaranth (Amaranthus hypochondriacus) meals positively affect plasma lipid profile in rats fed cholesterol-containing diets. AB - Cereals are an important part of diets for hypercholesterolemic patients. However, some of these patients are allergic to these natural products. The purpose of the current study was to compare oatmeal with equal in nutritional values two allergy-free amaranth meals to determine whether this pseudocereal can be a substitute for allergic to cereals individuals. The total phenols of the samples were determined with the Folin-Chocalteu reagent, anthocyanins, and flavonoids spectrophotometrically. The antioxidant activities were estimated with nitric oxide scavenging radical (NO) and by beta-carotene bleaching (beta carotene). It was found that the contents of different protein fractions, antioxidant compounds, and the antioxidant activities of oatmeal were significantly higher than those of the two amaranth samples. The results of kinetic reactions showed that samples differed in their capacities to quench these radicals, and oats have shown more antioxidant activity than amaranth. High correlation was observed between antioxidant activities and phenols (R(2) = 0.99). In the in vivo part of the investigation, 60 male Wistar rats were divided into five diet groups of 12 animals each; these groups were designated as Control, Chol, Chol/Oat, Chol/AmarI, and Chol/AmarII. The rats of the Control group were fed basal diet (BD) only. To the BD of the four other groups were added the following: 1% of cholesterol (Chol), 10% of oat meal and 1% of cholesterol (Chol/Oat), 10% of amaranth I meal, and 1% of cholesterol (Chol/AmarI) and 10% of amaranth II meal and 1% of cholesterol (Chol/AmarII). After 32 days of different feeding, diets supplemented with oat meal and, to lesser degree, with amaranth I and amaranth II hindered the rise in the plasma lipids: a) TC: 3.14 vs. 4.57 mmol/L, - 31.3%; 3.31 vs. 4.57 mmol/L - 27.6%; and 3.40 vs. 4.57, - 25.6%, respectively b) LDL-C: 1.69 vs. 3.31 mmol/L, - 49.9%; 2.05 vs. 3.31 mmol/L, - 38.1%; and 2.16 vs. 3.31 mmol/L, - 34.8%, respectively; c) TG: 0.73 vs. 0.88 mmol/L, - 17.1%; 0.75 vs. 0.88 mmol/L, - 14.8%; and 0.79 vs. 0.88 mmol/L, -10.2%, respectively. The HDL-PH was increased as follows: 0.79 vs. 0.63 mmol/L, -25.3%; 0.75 vs. 0.63 mmol/L, -23.0%; and 0.71 vs. 0.63 mmol/L, 12.7% for the Chol/Oat, Chol/AmarI and Chol/AmarII, respectively. No significant changes in the concentrations of HDL-C and TPH were found; however the HDL-C in the Chol/Oat group was slightly higher than in other groups. No changes in the Control group were registered. In conclusion, oat and amaranth meals positively affect plasma lipid profile in rats fed cholesterol-containing diets. The degree of this positive influence is directly connected to the contents of the bioactive components and the antioxidant activities of the studied samples. It is suggested that amaranth could be a valuable substitute for hypercholesterolemic patients allergic to cereals. PMID- 15542355 TI - Age-associated changes in intestinal fructose uptake are not explained by alterations in the abundance of GLUT5 or GLUT2. AB - A reduction in nutrient absorption may contribute to malnourishment in the elderly. The objectives of this study were to determine the effects of aging on the absorption of fructose in rats, as well as the mechanisms of these adaptive changes. Male Fischer 344 rats aged 1, 9, and 24 months were fed standard Purina chow for 2 weeks (PMI #5001, PMI Nutritionals, Brentwood, MO). The uptake of (14)C-labeled D-fructose was determined in vitro using the intestinal sheet method. Intestinal samples were taken for RNA isolation and for brush border membrane (BBM) and basolateral membrane (BLM) preparation. Northern blotting, Western blotting, and immunohistochemistry were used to determine the effects of age and diet on GLUT5 and GLUT2. When expressed on the basis of intestinal or mucosal weights, aging was associated with a decline in jejunal and ileal fructose uptake, whereas jejunal fructose uptake was increased when expressed on the basis of serosal or mucosal surface area. The alterations in fructose uptake were not paralleled by changes in GLUT5 or GLUT2 abundance. These results indicate that 1) the effect of age on fructose uptake depends on the method used to express results, and 2) the age-associated changes in uptake are not explained by alterations in GLUT5 and GLUT2. PMID- 15542356 TI - Influence of co-administration of oral insulin and docosahexaenoic acid in mice. AB - Insulin and docosahexaenoic acid are both present in human milk. The aim of this study was to examine the effect of co-administration of oral insulin and DHA in mice. Immediately after weaning, Balb C mice were divided into four groups of seven mice each for a period of 4 weeks. Group 1 received a chow diet only. Group 2 received a chow diet and also was given human insulin (1 unit/mL of drinking water) without docosahexaenoic acid. Group 3 received a chow diet supplemented with docosahexaenoic acid (500 mg/kg/day in the chow) and no insulin. Group 4 received a chow diet and supplementation with both human insulin and docosahexaenoic acid. At 28 days, fasting blood levels of glucose, insulin, lipids, lipid peroxidation analysis, docosahexaenoic acid plasma levels, and docosahexaenoic acid content in red blood cells were determined. We found that glucose levels were lower in the group that was supplemented with insulin only (group 2, 61.4 mg/dL +/- 2.8,mean +/- SD) and in the group that was supplemented with DHA only (group 3, 61.1 mg/dL +/- 2.0) compared to controls (group 1, 71 mg/dL +/- 6.9, P < 0.0001). Supplementation of both insulin and docosahexaenoic acid (group 4) resulted in significantly lower glucose levels (56.4 mg/dL +/- 2.6) compared to those in groups 2 and 3 (P < 0.01). No significant differences were found in lipid profile or lipid peroxidation between the groups. We conclude that adding insulin or docosahexaenoic acid to the diet of weaned Balb C mice reduces glucose blood levels. Supplementation with both substances has a synergistic effect. The presence of insulin and docosahexaenoic acid in human milk may be the cause for reduced glucose levels in breast-fed infants, in addition to the known effects of DHA on insulin sensitivity. PMID- 15542357 TI - Bacteriorhodopsin photocycle kinetics analyzed by the maximum entropy method. AB - A maximum entropy method (MEM) was developed for the study of the bacteriorhodopsin photocycle kinetics. The method can be applied directly to experimental kinetic absorption data without any assumption for the number of the intermediate states taking part in the photocycle. Though this method does not give a specific kinetics, its result is very useful for selection between possible photocycle kinetics. Using simulated data, it is shown that MEM gives correct results for the number of the intermediate states and the amplitude distributions around the characteristic lifetimes. Analyzing experimental absorption data at five different wavelengths, MEM gives seven or eight characteristic lifetimes, which means that at least so many distinct intermediate states exist during the photocycle. Many possible photocycle kinetic models were studied and compared with the MEM result. The best agreement was found with a branching photocycle model of eight intermediate states (K, L, M(1), M(2), M(3), M(4), N, O). The branching occurs at the L intermediate state (M(1) and M(2) being in one branch and M(3) and M(4) in the other branch), but at high pH it occurs already at the K state. PMID- 15542358 TI - Efficacy of different targeting agents in the photolysis of interleukin-2 receptor bearing cells. AB - The multichain interleukin-2 receptor (IL-2R) has been proposed as a target for immunotherapy in the treatment of certain cancers including adult T-cell leukemia and cutaneous T-cell lymphoma as well as certain autoimmune diseases. The IL-2R is abnormally expressed on cells associated with each of these diseases; while normal, non-activated T-cells do not express the receptor. This report describes the selective photolysis of activated and non-activated IL-2R expressing cells using several immunoconjugates synthesized with one of two photosensitizers, hematoporphyrin (HP) or chlorin-e(6) (Ce(6)), covalently linked to IL-2 or an anti-IL-2R antibody. Destruction of IL-2R bearing cells was achieved after photosensitizer internalization and irradiation using all tested photosensitizer conjugates. Chlorin containing conjugates were more effective, by a factor of 4 or more, than HP containing conjugates. Conjugates made with IL-2 were up to 30 times more effective than conjugates that used a monoclonal antibody against the IL-2R for targeting. Activation of the cells to increase IL-2R expression decreased the internalization time required for optimal therapeutic efficacy; however, stimulation of the cell to increase IL-2 secretion greatly reduced conjugate effectiveness. This work could lead to the development of more effective strategies to treat T-cell diseases. PMID- 15542359 TI - Effect of UVA radiation on membrane fluidity and radical decay in human fibroblasts as detected by spin labeled stearic acids. AB - The ultraviolet A (UVA) radiation component of sunlight (320-400 nm) has been shown to be a source of oxidative stress to cells via generation of reactive oxygen species. We report here some consequences of the UVA irradiation on cell membranes detected by electron paramagnetic resonance (EPR) spectroscopy. Paramagnetic nitroxide derivatives of stearic acid bearing the monitoring group at different depths in the hydrocarbon chain were incorporated into human fibroblasts membranes to analyze two main characteristics: kinetics of the nitroxide reduction and membrane fluidity. These two characteristics were compared for control and UVA-irradiated (0-250 kJ/m(2)) cells. The term relative redox capacity (RRC) was introduced to characterize and to compare free radical reduction measured by EPR with some well-known viability/clonogenicity tests. Our results showed that UVA-irradiation produces a more rigid membrane structure, especially at higher doses. Furthermore, we found that trends agree in survival measured by neutral red (NR), trypan blue (TB), and clonogenic efficiency compared with RRC values measured by EPR for low and medium exposure doses. Above 100 kJ/m(2), differences between these tests were observed. Antioxidant effect was modeled by alpha-tocopherol-acetate treatment of the cells before UVA irradiation. While NR, TB and clonogenicity tests showed protection at the highest UVA doses (>100 kJ/m(2)), results obtained with EPR measurements, both membrane fluidity and kinetics, or using MTT test did not exhibit this protective effect. PMID- 15542360 TI - Inactivation of dengue virus by methylene blue/narrow bandwidth light system. AB - Peracetic acid was one of the most commonly used disinfectants on solid surfaces in hospitals or public places. However, peracetic acid is an environmental toxin. Therefore, safer, alternative disinfectants or disinfectant systems should be developed. Because photodynamic virus inactivation with methylene blue (MB)/light system has proven effective in blood banking, MB was selected as a photosensitizing agent, dengue virus as a model virus for enveloped RNA viruses, and an in-house fabricated narrow bandwidth light system overlapping the absorption spectrum of MB as the light source. Dengue virus was mixed with different concentrations of MB, and illuminated by the narrow bandwidth light system under different illumination distances and times. The amount of dengue virus remaining was evaluated by plaque forming assays. Results showed that the concentration of MB working solution, illumination intensity of light source, illumination distance and time were four key factors affecting efficiency of virus inactivation using the MB/narrow bandwidth light system. Dengue virus could be completely inactivated at 2.5 m in 5 min when MB >/= 1.0 microg/ml. However, when the distance reached 3.0 m, only greater concentrations of MB (2.0 microg/ml) could completely inactivate virus in a reasonably short time (20 min), and smaller concentrations of MB (1.0 microg/ml) could only completely inactivate virus using longer times (25 min). The results of this virus inactivation model indicate that our MB/narrow bandwidth light system provides a powerful, easy way to inactivate dengue viruses. PMID- 15542361 TI - DNA ligand Hoechst-33342 enhances UV induced cytotoxicity in human glioma cell lines. AB - The effects of minor groove binding ligand bisbenzimidazole derivative Hoechst 33342 on the cellular response to UV damage have been studied in two human glioma cell lines BMG-1 and U-87 grown as monolayer cultures. Treatment induced cell death (macro-colony assay) and growth inhibition, potential lethal damage recovery, cytogenetic damage (micronuclei formation) and proliferation kinetics were studied as parameters for cellular response. Pre and post-irradiation treatment with Hoechst-33342 (1-20 microM) enhanced the UV-induced growth inhibition and cell death in a concentration dependent manner in both cell lines. At higher Hoechst-33342 concentrations (>5 microM), the cytotoxic effects of the combination (Hoechst-33342+UV) were highly synergistic and mainly mediated through apoptosis implying the possible interactions of lesions caused by both the agents. The enhanced cell death due to Hoechst-33342 was accompanied by a significant increase (2-3 folds at 5 microM) in UV-induced micronuclei formation in BMG-1 cells. Under these conditions, Hoechst-33342 also enhanced the UV induced cell cycle delay, mainly due to S and G(2) blocks. The increase in UV induced micronuclei formation observed after treatment with Hoechst-33342 indicates that the DNA bound Hoechst-33342 may interfere with the rejoining of DNA strand breaks. Since the treatment of cells with the replication inhibitor aphidicolin reduced the enhancement of UV induced cytotoxicity by Hoechst-33342, ongoing DNA replication appears to stimulate Hoechst-33342 and UV-induced cytotoxicity. PMID- 15542362 TI - Near-ultraviolet photolysis of L-mandelate, formation of reactive oxygen species, inactivation of phage T7 and implications on human health. AB - Compared with ultraviolet B and C, UVA is considered to have little direct effects on biological systems. However, damaging effects of UVA on biological systems are often synergistically enhanced in the presence of sensitizers. Production of reactive oxygen species (ROS) has been implicated in the process. Several ROS have been identified but their involvement in inducing cellular damage is yet to be fully evaluated. Although membranes and proteins are affected, DNA is an important target and a variety of types of damage have been reported. Here, we present evidence that L-mandelate can act as a near UV (NUV) sensitizer, when activated by a lamp emitting 99% UVA and 1% UVB. Although evidence is available that H(2)O(2) and a small amount of *OH are produced, an alternative effect of the sensitization reaction may involve direct electron transfer. Studies have shown that NUV photolysis of mandelate can inactivate phage T7. Employment of tetrazolium blue test to detect superoxide anion may not be sufficient evidence as this agent may be reduced by alternative routes. PMID- 15542363 TI - Light-induced modulation of Porphyromonas gingivalis growth. AB - The bacterium Porphyromonas gingivalis is a clinically significant agent in periodontitis, a disease for which there is no definitive cure. Several groups have attempted to kill this bacterium using low levels of light in the absence of a photosensitizer, with conflicting results. We hypothesize that it is not possible to kill P. gingivalis by targeting endogenous porphyrins for a photochemical reaction. We demonstrated that irradiation of P. gingivalis with 455 or 625 nm light emitting diodes did not induce a photochemical killing of the cultures. Controlled temperature experiments indicate that irradiation at either wavelength did not significantly impact the growth of P. gingivalis cultures, as compared to non-irradiated controls. Rather, the irradiation caused a temperature increase in the growth medium, which altered the growth of the cultures. These results indicate that heat-induced killing of P. gingivalis could be the mechanism behind successful irradiation experiments with this bacterium. PMID- 15542364 TI - UVR reflections at the surface of the eye. AB - Sunlight plays an etiological role in the formation of skin cancers [Phys. Med. Biol. 24 (1979) 931]. Non-melanoma skin cancers commonly arise in sun-exposed parts of the body, especially on the head and neck regions [Int. J. Dermatol. 34 (6) (1995) 398] although the amount of sun exposure that is required for the formation of skin cancers is still unknown. It is known that the larger the dose of UVR, and in particular the erythemal action spectrum, the more likely it is to form the non-melanoma skin cancers, basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs). A number of regions on the face exhibit a seemingly high rate of occurrence of BCCs in relation to apparent direct exposure. One of these regions is the inner canthus located next to the eye on the bridge of the nose which has an occurrence rate of 7.1% of all BCCs that occur on the head and neck [Gen. Surg. 51 (6) (1981) 576, Aust. NZ J. Surg. 60 (1990) 855, Malignant Skin Tumours, Longman Singapore Publishers, Singapore, 1991]. The inner canthus seems to be well protected from large direct doses of ultraviolet radiation (UVR) and to explain the higher incidence of BCCs on the inner canthus it is proposed that a significant proportion of the incident UVR on the eye and surrounding areas is reflected onto the inner canthus. This paper presents a preliminary investigation of the contribution of UVR reflected to the inner canthus from the tear film covering the eye using a two-dimensional theoretical model on the horizontal plane (0 degrees elevation angle). Calculations show that up to 30% of the total radiation that is received on the inner canthus on a cellular level in this plane is reflected from the eye. A three-dimensional computer-generated ray tracing model of the eye, surrounding facial features and the inner canthus is being created to investigate the effect that these reflections have on the total dose of UVR. PMID- 15542365 TI - The action of oxygen on chlorophyll fluorescence quenching and absorption spectra in pea thylakoid membranes under the steady-state conditions. AB - The effect of oxygen concentration on both absorption and chlorophyll fluorescence spectra was investigated in isolated pea thylakoids at weak actinic light under the steady-state conditions. Upon the rise of oxygen concentration from anaerobiosis up to 412 microM a gradual absorbance increase around both 437 and 670 nm was observed, suggesting the disaggregation of LHCII and destacking of thylakoids. Simultaneously, an increase in oxygen concentration resulted in a decline in the Chl fluorescence at 680 nm to about 60% of the initial value. The plot of normalized Chl fluorescence quenching, F(-O(2))/F(+O(2)), showed discontinuity above 275 microM O(2), revealing two phases of quenching, at both lower and higher oxygen concentrations. The inhibition of photosystem II by DCMU or atrazine as well as that of cyt b(6)f by myxothiazol attenuated the oxygen induced quenching events observed above 275 microM O(2), but did not modify the first phase of oxygen action. These data imply that the oxygen mediated Chl fluorescence quenching is partially independent on non-cyclic electron flow. The second phase of oxygen-induced decline in Chl fluorescence is diminished in thylakoids with poisoned PSII and cyt b(6)f activities and treated with rotenone or N-ethylmaleimide to inhibit NAD(P)H-plastoquinone dehydrogenase. The data suggest that under weak light and high oxygen concentration the Chl fluorescence quenching results from interactions between oxygen and PSI, cyt b(6)f and Ndh. On the contrary, inhibition of non-cyclic electron flow by antimycin A or uncoupling of thylakoids by carbonyl cyanide m-chlorophenyl hydrazone did not modify the steady-state oxygen effect on Chl fluorescence quenching. The addition of NADH protected thylakoids against oxygen-induced Chl fluorescence quenching, whereas in the presence of exogenic duroquinone the decrease in Chl fluorescence to one half of the initial level did not result from the oxygen effect, probably due to oxygen action as a weak electron acceptor from PQ pool and an insufficient non photochemical quencher. The data indicate that mechanism of oxygen-induced Chl fluorescence quenching depends significantly on oxygen concentration and is related to both structural rearrangement of thylakoids and the direct oxygen reduction by photosynthetic complexes. PMID- 15542367 TI - From genome to vaccine--new immunoinformatics tools for vaccine design. PMID- 15542366 TI - Inhibition of immediate type hypersensitivity reaction by combined irradiation with ultraviolet and visible light. AB - Recently we found that ultraviolet B (UVB) irradiation in erythematous doses significantly inhibited the immediate type hypersensibility reaction in the skin. In the present study we investigated the effects of different wavelengths on the skin prick test reaction (SPT). The forearm of ragweed allergic patients was irradiated with increasing doses of ultraviolet A (UVA), visible light (VIS) or combined UVB, UVA and VIS light, referred to as mUV/VIS. SPTs were performed 24 h after irradiation both on irradiated and non-irradiated control skin areas using ragweed extract. UVA and VIS irradiation led to a slight, not significant inhibition of allergen-induced wheal formation. Mixed irradiation with mUV/VIS light resulted in a dose-dependent inhibition of the allergen-induced wheal formation. The inhibition was significant already at suberythematous doses. As there is a good correlation between SPT and the nasal symptoms in patients with hay fever these data suggest that phototherapy with mUV/VIS light might be an effective and safe treatment modality for immediate type hypersensibility reactions in the skin and nasal mucosa. PMID- 15542368 TI - Virtual models of the HLA class I antigen processing pathway. AB - Antigen recognition by cytotoxic CD8 T cells is dependent upon a number of critical steps in MHC class I antigen processing including proteosomal cleavage, TAP transport into the endoplasmic reticulum, and MHC class I binding. Based on extensive experimental data relating to each of these steps there is now the capacity to model individual antigen processing steps with a high degree of accuracy. This paper demonstrates the potential to bring together models of individual antigen processing steps, for example proteosome cleavage, TAP transport, and MHC binding, to build highly informative models of functional pathways. In particular, we demonstrate how an artificial neural network model of TAP transport was used to mine a HLA-binding database so as to identify HLA binding peptides transported by TAP. This integrated model of antigen processing provided the unique insight that HLA class I alleles apparently constitute two separate classes: those that are TAP-efficient for peptide loading (HLA-B27, -A3, and -A24) and those that are TAP-inefficient (HLA-A2, -B7, and -B8). Hence, using this integrated model we were able to generate novel hypotheses regarding antigen processing, and these hypotheses are now capable of being tested experimentally. This model confirms the feasibility of constructing a virtual immune system, whereby each additional step in antigen processing is incorporated into a single modular model. Accurate models of antigen processing have implications for the study of basic immunology as well as for the design of peptide-based vaccines and other immunotherapies. PMID- 15542369 TI - Computational methods for prediction of T-cell epitopes--a framework for modelling, testing, and applications. AB - Computational models complement laboratory experimentation for efficient identification of MHC-binding peptides and T-cell epitopes. Methods for prediction of MHC-binding peptides include binding motifs, quantitative matrices, artificial neural networks, hidden Markov models, and molecular modelling. Models derived by these methods have been successfully used for prediction of T-cell epitopes in cancer, autoimmunity, infectious disease, and allergy. For maximum benefit, the use of computer models must be treated as experiments analogous to standard laboratory procedures and performed according to strict standards. This requires careful selection of data for model building, and adequate testing and validation. A range of web-based databases and MHC-binding prediction programs are available. Although some available prediction programs for particular MHC alleles have reasonable accuracy, there is no guarantee that all models produce good quality predictions. In this article, we present and discuss a framework for modelling, testing, and applications of computational methods used in predictions of T-cell epitopes. PMID- 15542370 TI - Quantitative structure-activity relationships and the prediction of MHC supermotifs. AB - The underlying assumption in quantitative structure-activity relationship (QSAR) methodology is that related chemical structures exhibit related biological activities. We review here two QSAR methods in terms of their applicability for human MHC supermotif definition. Supermotifs are motifs that characterise binding to more than one allele. Supermotif definition is the initial in silico step of epitope-based vaccine design. The first QSAR method we review here--the additive method--is based on the assumption that the binding affinity of a peptide depends on contributions from both amino acids and the interactions between them. The second method is a 3D-QSAR method: comparative molecular similarity indices analysis (CoMSIA). Both methods were applied to 771 peptides binding to 9 HLA alleles. Five of the alleles (A*0201, A*0202, A*0203, A*0206 and A*6802) belong to the HLA-A2 superfamily and the other four (A*0301, A*1101, A*3101 and A*6801) to the HLA-A3 superfamily. For each superfamily, supermotifs defined by the two QSAR methods agree closely and are supported by many experimental data. PMID- 15542371 TI - A structure-based approach for prediction of MHC-binding peptides. AB - Identification of immunodominant peptides is the first step in the rational design of peptide vaccines aimed at T-cell immunity. The advances in sequencing techniques and the accumulation of many protein sequences without the purified protein challenge the development of computer algorithms to identify dominant T cell epitopes based on sequence data alone. Here, we focus on antigenic peptides recognized by cytotoxic T cells. The selection of T-cell epitopes along a protein sequence is influenced by the specificity of each of the processing stages that precede antigen presentation. The most selective of these processing stages is the binding of the peptides to the major histocompatibility complex molecules, and therefore many of the predictive algorithms focus on this stage. Most of these algorithms are based on known binding peptides whose sequences have been used for the characterization of binding motifs or profiles. Here, we describe a structure-based algorithm that does not rely on previous binding data. It is based on observations from crystal structures that many of the bound peptides adopt similar conformations and placements within the MHC groove. The algorithm uses a structural template of the peptide in the MHC groove upon which peptide candidates are threaded and their fit to the MHC groove is evaluated by statistical pairwise potentials. It can rank all possible peptides along a protein sequence or within a suspected group of peptides, directing the experimental efforts towards the most promising peptides. This approach is especially useful when no previous peptide binding data are available. PMID- 15542372 TI - Candidate epitope identification using peptide property models: application to cancer immunotherapy. AB - Peptides derived from pathogens or tumors are selectively presented by the major histocompatibility complex proteins (MHC) to the T lymphocytes. Antigenic peptide MHC complexes on the cell surface are specifically recognized by T cells and, in conjunction with co-factor interactions, can activate the T cells to initiate the necessary immune response against the target cells. Peptides that are capable of binding to multiple MHC molecules are potential T cell epitopes for diverse human populations that may be useful in vaccine design. Bioinformatical approaches to predict MHC binding peptides can facilitate the resource-consuming effort of T cell epitope identification. We describe a new method for predicting MHC binding based on peptide property models constructed using biophysical parameters of the constituent amino acids and a training set of known binders. The models can be applied to development of anti-tumor vaccines by scanning proteins over-expressed in cancer cells for peptides that bind to a variety of MHC molecules. The complete algorithm is described and illustrated in the context of identifying candidate T cell epitopes for melanomas and breast cancers. We analyzed MART-1, S 100, MBP, and CD63 for melanoma and p53, MUC1, cyclin B1, HER-2/neu, and CEA for breast cancer. In general, proteins over-expressed in cancer cells may be identified using DNA microarray expression profiling. Comparisons of model predictions with available experimental data were assessed. The candidate epitopes identified by such a computational approach must be evaluated experimentally but the approach can provide an efficient and focused strategy for anti-cancer immunotherapy development. PMID- 15542373 TI - Discovery of promiscuous HLA-II-restricted T cell epitopes with TEPITOPE. AB - TEPITOPE is a prediction model that has been successfully applied to the in silico identification of T cell epitopes in the context of oncology, allergy, infectious diseases, and autoimmune diseases. Like most epitope prediction models, TEPITOPE's underlying algorithm is based on the prediction of HLA-II peptide binding, which constitutes a major bottleneck in the natural selection of epitopes. An important step in the design of subunit vaccines is the identification of promiscuous HLA-II ligands in sets of disease-specific gene products. TEPITOPE's user interface enables the systematic prediction of promiscuous peptide ligands for a broad range of HLA-binding specificity. We show how to apply the TEPITOPE prediction model to identify T cell epitopes, and provide both a road map and examples of its successful application. PMID- 15542374 TI - Engineering immunogenic consensus T helper epitopes for a cross-clade HIV vaccine. AB - Developing a vaccine that will stimulate broad HIV-specific T cell responses is difficult because of the variability in HIV T cell epitope sequences, which is in turn due to the high mutation rate and consequent strain diversity of HIV-1. We used a new Class II version of the EpiMatrix T cell epitope-mapping tool and Conservatrix to select highly conserved and promiscuous Class II HLA-restricted T cell epitopes from a database of 18,313 HIV-1 env sequences. Criteria for selection were: (1) number of HIV-1 strains represented as measured by Conservatrix; (2) EpiMatrix score; and (3) promiscuity (number of unique MHC motifs contained in the peptide). Using another vaccine design tool called the EpiAssembler, a new set of overlapping, conserved and immunogenic HIV-1 peptides were engineered creating extended "immunogenic consensus" sequences. Each overlapping 9-mer of the 20-23 amino acid long immunogenic consensus peptides was conserved in a large number (range 893-2254) of individual HIV-1 strains, although the novel peptides were not representative of any single strain of HIV. We synthesized nine representative peptides. T helper cell responses to the peptides were evaluated by ELISpot (gamma-interferon) assay, using peripheral blood monocytes (PBMC) obtained from 34 healthy long term non-progressor (LT) or moderate-progressor (MP) donors (median years infected = 8.88, median CD4 T cells = 595, median VL = 1044). Nine peptides were tested, of which eight were confirmed in ELISpot assays using PBMC from the LT/MP subjects. These epitopes were ranked by Conservation and EpiMatrix score 1, 2, 3, 5, 7, 11, and 14 out of the set of 9 original peptides. Five of these peptides were selected for inclusion in an epitope-driven cross-clade HIV-1 vaccine (the GAIA vaccine). These data confirm the utility of bioinformatics tools to select and construct novel "immunogenic consensus sequence" T cell epitopes for a globally relevant vaccine against HIV. PMID- 15542375 TI - Textmining in support of knowledge discovery for vaccine development. AB - Complete genome data of infectious microorganisms permit systematic computational sequence-based predictions and experimental testing of candidate vaccine epitopes. Both, predictions and the interpretation of experiments rely on existing information in the literature which is mostly manually extracted and curated. The growing amount of data and literature information has created a major bottleneck for the interpretation of results and maintenance of curated databases. The lack of suitable free-text information extraction, processing, and reporting tools prompted us to develop a knowledge discovery support system that enhances the understanding of immune response and vaccine development. The current prototype system, Gene expression/epitpopes/protein interaction (GEpi), focuses on molecular functions of HIV-infected T-cells and HIV epitope information, using textmining, and interrelation of biomolecular data from domain specific databases with MEDLINE abstract-inferred information. Results showed that extraction and processing of molecular interaction, disease associations, and gene ontology-derived functional information generate intuitive knowledge reports that aid the interpretation of host-pathogen interaction. In contrast, epitope (word and sequence) information in MEDLINE abstracts is surprisingly sparse and often lacks necessary context information, such as HLA-restriction. Since the majority of epitope information is found in tables, figures, and legends of full-text articles, its extraction may not require sophisticated natural language processing techniques. Support of vaccine development through textmining requires therefore the timely development of domain-specific extraction rules for full-text articles, and a knowledge model for epitope related information. PMID- 15542376 TI - Contribution of vasoconstriction to the origin of atherosclerosis: a conceptual study. AB - Research during the past century has clearly shown that endothelial injury (EI) and/or endothelial dysfunction (ED) are among the major events determining the onset of atherosclerosis. Included in the events that may elicit endothelial damage, vasoconstriction (VC) has received relatively little attention. This conceptual review attempts to show that in elastic and conduit arteries, VC is not only capable of producing EI/ED, but is also closely associated with many recognized proatherogenic stimuli. Of related interest is the observation that a number of suspected antiatherogenic stimuli oppose VC by their vasodilatory effects, lending further support to this relationship. In addition, recent developments in the knowledge of the molecular basis of VC (including the role of specific inhibitors) are discussed, and their potential for preventing lesion formation and thus becoming novel therapeutic alternatives against the onset of atherosclerosis are highlighted. PMID- 15542377 TI - Autologous cell-based therapies for vascular disease. AB - Strategies that enhance the number of endothelial cells (ECs) in the vessel wall following injury may limit complications such as thrombosis, vasospasm, and neointimal formation through reconstitution of a luminal barrier and cellular secretion of paracrine factors. Proof of principle has been demonstrated by studies in which mature ECs, culture expanded from harvested vascular tissue, were seeded in the arterial wall following balloon injury. The recent identification of circulating cells capable of developing an endothelial phenotype, including progenitor cells, has raised the possibility of using blood derived cells as therapeutic agents. This article reviews data suggesting that such cells confer vascular protective effects after injury, raising the potential for novel, autologous approaches to the treatment of vascular disease. PMID- 15542378 TI - iNOS gene transfer for graft disease. AB - Vascular bypass surgery involves the use of a vascular conduit to circumvent a site of vascular compromise. Vascular graft failure continues to plague both the patients receiving and the surgeons performing these interventions. Demand for the development of a therapy to reduce intimal hyperplasia--the most common cause of bypass graft failure--is significant and has been the goal of many biotechnology groups. The development of gene therapy as a feasible clinical intervention has allowed for novel methods of inhibiting intimal hyperplasia to be conceived. This review describes the evolution of gene transfer of the inducible nitric oxide synthase (iNOS) gene, one of the most successful preclinical interventions to date for vascular disease. PMID- 15542379 TI - Selenoproteins, cholesterol-lowering drugs, and the consequences: revisiting of the mevalonate pathway. AB - 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) and peroxisome proliferator-activated receptor alpha activators (fibrates) are the backbone of pharmacologic hypercholesterolemia and dyslipidemia treatment. Many of their clinical effects, however, are still enigmatic. This article describes how a side road of the mevalonate pathway, characterized in recent years, can rationalize a major fraction of these unexplained observations. This side road is the enzymatic isopentenylation of selenocysteine-tRNA([Ser]Sec) (Sec-tRNA), the singular tRNA to decode the unusual amino acid selenocysteine. The functionally indispensable isopentenylation of Sec-tRNA requires a unique intermediate from the mevalonate pathway, isopentenyl pyrophosphate, which concomitantly constitutes the central building block for cholesterol biosynthesis, and whose formation is suppressed by statins and fibrates. The resultant inhibition of Sec tRNA isopentenylation profoundly decreases selenoprotein expression. This effect might seamlessly explain the immunosuppressive, redox, endothelial, sympatholytic, and thyroidal effects of statins and fibrates as well as their common side effects and drug interactions. PMID- 15542381 TI - Smooth muscle progenitor cells in vascular disease. AB - Vascular smooth muscle cells have long been described as phenotypically diverse, with some populations possessing the potential for monoclonal expansion within the adult vessel wall. Recent experimental and human data suggest a potential role in atherosclerotic plaque, transplant arteriopathy, and angiogenesis biology for putative smooth muscle progenitor cells in the circulation, adult tissues, and the perivascular adventitia. The location, mobilization, and function of such progenitors is still poorly understood, but may include reparative or alternatively proliferative consequences for plaque development and progression. These observations provide a platform for re-evaluation of smooth muscle cell heterogeneity and progenitor biology in atherosclerosis, with implications for diagnosis and therapy of vascular disease. PMID- 15542380 TI - Genomic and proteomic profiles of heart disease. AB - Genomics and proteomics are becoming powerful tools for profiling diseased states. The human genome is estimated to encode 30,000 to 40,000 genes, generating more than 100,000 functionally distinct proteins. Microarray data are available for multiple models of heart disease as well as for diseased and failing human hearts. Similarly, two-dimensional gel data banks of normal and diseased myocardium from multiple species are published and are available on the Internet. The combined technologies are beginning to provide new insights into the causes and pathways of cardiac dysfunction. This article reviews the novel findings that have been acquired from genomic and proteomic screens of diseased hearts in animal models and humans. PMID- 15542382 TI - MR imaging of the neonatal brain at 3 Tesla. AB - 3 Telsa MR scanners are now becoming more widely available and 3 Telsa is likely to become the filed strength of choice for clinical imaging of the brain. The neonatal brain can be safely and successfully imaged at 3 Telsa. The improved signal to noise afforded by a higher field strength may be used to improve image quality or shorten acquisition times. This may be exploited for conventional T1 and T2 weighted imaging and also for advanced techniques such as diffusion tensor imaging, angiography and functional magnetic resonance studies. PMID- 15542383 TI - Behavioural and neuropsychological problems in refractory paediatric epilepsies. AB - The associated behavioural and neuropsychological profiles were studied in 573 children with refractory epilepsy, admitted in our residential Rehabilitation and Epilepsy Unit during the period 1984-2000. The aim of this study was to look for possible correlations between epileptic categories and behavioural profiles. The most frequent neuro-behavioural correlates in the different epilepsy categories were pervasive disorders (48/573=8%), attentional problems (43/573=7.5%), loss of self-esteem (n=49 or 9%) and self-induction of seizures (n=34 or 7%). Pervasive disorders were significantly more frequent in secondary generalised epilepsies. In 86/573 children (15%), mental decline due to epileptic process itself was observed. As expected this was seen in all patients with Lennox-Gastaut syndrome, West syndrome, severe myoclonic epilepsy of infancy and in continuous spikes and slow waves during slow wave sleep. The only behavioural problem that was more frequent in females was self-induction of seizures. PMID- 15542384 TI - Analysis of the mitochondrial encoded subunits of complex I in 20 patients with a complex I deficiency. AB - NADH-ubiquinone oxidoreductase or complex I deficiency is a frequently diagnosed enzyme defect of the oxidative phosphorylation (OXPHOS) system in humans. However, in many patients, with complex I deficiency and clinical symptoms suggestive of mitochondrial disease, often no genetic defect can be found after investigation of the most common mitochondrial DNA (mtDNA) mutations. In this study, 20 patients were selected with a biochemically documented complex I defect and no common mtDNA mutation. We used the Denaturing Gradient Gel Electrophoresis (DGGE) method with primers encompassing all mitochondrial encoded fragments, to search in a systematic manner for mutations in the mitochondrial genome of complex I. In our group of patients, we were able to detect a total of 96 nucleotide changes. We were not able to find any disease causing mutation in the mitochondrial encoded subunits of complex I. These results suggested that the complex I deficiency in this group of patients is most probably caused by a defect in one of the nuclear encoded structural genes of complex I, or in one of the genes involved in proper assembly of the enzyme. PMID- 15542385 TI - Recurrent cerebral fever in the seventeenth and twenty first centuries. AB - The development of modern neuroscience away from the concepts of Hippocrates and Galen can be traced to the writings of some 17th century clinicians, especially Thomas Willis. His exceptional skills in observation and description allow a window into the experiences of our medical forebears. His approach to the management of infection-related coma in a child is amenable to modern interpretation and comparison with modern management because of the clarity of his clinical descriptions. Modern clinicians may benefit from this historical perspective into influences on the origins of neuroscience. The different outcome for a child presenting in the 17th and 21st century encourage grateful reflection on our current privileged position. PMID- 15542386 TI - A severe case of facioscapulohumeral muscular dystrophy (FSHD) with some uncommon clinical features and a short 4q35 fragment. AB - Severe and early facioscapulohumeral muscular dystrophy (FSHD) is relatively rare. In this report, we describe a case of severe, infantile onset FSHD in a patient with asymmetric progressive ptosis and early hyperlordosis. DNA analysis revealed a very short 4q35 allele of 8.6 kb and a somatic mosaicism for the 4q35 deletion detected in a subclinically affected parent of the patient. This case demonstrates difficulties in the management of abnormal posture, especially early hyperlordosis in children with this disorder. PMID- 15542387 TI - Progressive encephalopathy with edema, hypsarrhythmia and optic atrophy (PEHO) syndrome in a Swiss child. AB - Progressive encephalopathy with edema, hypsarrhythmia and optic atrophy (PEHO) syndrome is a rare neurodegenerative syndrome first reported in 1991. Most patients described are of Finnish descent and very few patients have been reported in other countries. We report the first Swiss patient who fulfils the criteria of the PEHO syndrome. The course of the epilepsy is less severe than previously reported. Our patient developed a severe dystonic state after a febrile gastrointestinal infection, with a hypotonic state that may have been a dysregulation of brainstem origin. The diagnosis was made because of marked cerebellar atrophy in the repeated MRI. In patients with infantile spasms and severe developmental delay PEHO syndrome should be considered; it is not confined to Finnish heritage. Optic atrophy should be looked for and repeat MRI is indicated. PMID- 15542388 TI - The influence of phenobarbital overdose on aEEG recording. AB - Monitoring infants with amplitude integrated electroencephalography (aEEG) in neonatal units is becoming widespread. A case of a preterm infant who was monitored with aEEG due to suspected seizures is presented. The case emphasizes both the usefulness and drawback of aEEG. An erroneous diagnosis of seizure lead to phenobarbital overdose and the change of aEEG voltage and pattern assisted in a prompt diagnosis. PMID- 15542389 TI - Autism in tuberous sclerosis. AB - Despite considerable progress in the last few years, the neurobiologic basis of autism in tuberous sclerosis complex is still largely unknown and its clinical management represents a major challenge for child neurologists. Recent evidence suggests that early-onset refractory epilepsy and functional deficits associated with the anatomical lesions in the temporal lobes may be associated with autism. No one factor alone (cognitive impairment, tuber localization, occurrence of infantile spasms, focal EEG abnormalities), can be causally linked with the abnormal behaviour. Autism may also reflect a direct effect of the abnormal genetic program. Incidence of autism associated with Tuberous Sclerosis may be significantly higher than the rates of cardiac and renal abnormalities, for which screening is routinely conducted in this population. Hopefully, early diagnosis of autism will allow for earlier treatment and the potential for better outcome for children with Tuberous Sclerosis. PMID- 15542390 TI - Phenotype of non-syndromic deafness associated with the mitochondrial A1555G mutation is modulated by mitochondrial RNA modifying enzymes MTO1 and GTPBP3. AB - Phenotypic expression of the deafness-associated mitochondrial A1555G mutation in the 12S rRNA gene is influenced by aminoglycosides and complex inheritance of nuclear-encoded modifier genes. The position of a major nuclear modifier gene has been localized to chromosome 8p23.1, but the identification of this gene has remained elusive. Recently, we identified a second modifier gene, mitochondrial transcription factor B1 (TFB1M), involved in mitochondrial rRNA modification. In the present study, we tested three genes involved in mitochondrial tRNA or rRNA modification, and two genes associated with non-syndromic deafness, for linkage and linkage disequilibrium (LD) in 214 DNA samples from Spanish, Italian, and Arab-Israeli families with maternally inherited non-syndromic hearing loss. The multipoint non-parametric linkage analysis and transmission disequilibrium test testing were done using all families combined as well as divided based on linkage to the chromosome 8 locus and ethnicity. Two genes, MTO1 and GTPBP3, showed strongly suggestive linkage and significant LD results. Since both genes, as well as TFB1M, are involved in the process of mitochondrial RNA modification, it appears that the modification of mitochondrial RNA is an important regulatory pathway in the phenotypic expression of the deafness-associated mitochondrial A1555G mutation. This conclusion was supported by comparing linkage results of simulated genotypes with actual results for the four genes involved in mitochondrial RNA modification. PMID- 15542391 TI - Phenylalanine loading as a diagnostic test for DRD: interpreting the utility of the test. AB - Phenylalanine loading has been proposed as a diagnostic test for autosomal dominant DRD (dopa-responsive dystonia), and recently, a phenylalanine/tyrosine (phe/tyr) ratio of 7.5 after 4 h was reported as diagnostic of DRD. To test the utility of this test in another sample with DRD, we administered an oral challenge of phenylalanine (100 mg/kg) to 11 individuals with DRD and one non manifesting gene carrier. Only 6/12 had a 4 h phe/tyr ratio of greater than 7.5, suggesting that additional parameters must be set to avoid missing the diagnosis of DRD, including the need for the plasma phenylalanine to reach a minimum level 600 in order for the test to be valid. We propose that in cases where this minimum plasma phenylalanine level is not reached, plasma tetrahydrobiopterin should be measured or alternatively other symptomatic family members should be screened. PMID- 15542392 TI - Clinical heterogeneity of neonatal intrahepatic cholestasis caused by citrin deficiency: case reports from 16 patients. AB - A deficiency of citrin, which is encoded by the SLC25A13 gene, causes both adult onset type II citrullinemia (CTLN2) and neonatal intrahepatic cholestasis (NICCD). We analyzed 16 patients with NICCD to clarify the clinical features of the disease. Severe intrahepatic cholestasis with fatty liver was the most common symptom, but the accompanying clinical features were variable, namely; suspected cases of neonatal hepatitis or biliary atresia, positive results from newborn screening, tyrosinemia, failure to thrive, hemolytic anemia, bleeding tendencies and ketotic hypoglycemia. Laboratory data showed elevated serum bile acid levels, hypoproteinemia, low levels of vitamin K-dependent coagulation factors, and hypergalactosemia. Hypercitrullinemia was detected in 11 out of 15 patients examined. Most of the patients were given a lactose-free and/or medium chain triglycerides-enriched formula and lipid-soluble vitamins. The prognosis of the 16 patients is going fairy well at present, but we should observe these patients carefully to see if they manifest any symptom of CTLN2 in the future. PMID- 15542393 TI - Niemann-Pick type C disease: importance of N-glycosylation sites for function and cellular location of the NPC2 protein. AB - Niemann-Pick disease type C (NPC), a neurovisceral disorder characterized by accumulation of cholesterol and glycolipids in the lysosomal/late endosomal system, is due to mutations on either the NPC1 or the NPC2 genes. Although NPC1 and NPC2 proteins appear essential for proper cellular cholesterol trafficking, their precise functions and relationship have remained elusive. Mutation identification in NPC2 patients did not provide insights into structure-function relationships, but recent studies brought important information on the cholesterol-binding site of the NPC2 protein. The present work was focused on localization and N-glycosylation of NPC2, considering that glycosylation is often essential for targeting, stability and biological function of proteins. Using immunocytofluorescence in cultured human fibroblasts, we found that the native NPC2 protein is essentially lysosomal, at variance with the late endosomal location of NPC1. Expression of cDNA mutants affecting each of the three potential NPC2 N-glycosylation sites in NPC2-/- fibroblasts showed that only two sites are used. The intracellular human NPC2 protein occurred as two N glycosylated forms, with either one single oligosaccharide chain attached to Asn 58 or two oligosaccharides attached to Asn 58 and 135. The oligosaccharidic chains were of the hybrid and/or high mannose type, with no complex chains. Further studies on the cellular location of Asn 58 and Asn 135 mutant proteins and their respective effect on restoration of normal cholesterol traficking in NPC2-/- cells led to the conclusion that only the oligosaccharide chain carried by Asn 58 is responsible for proper targeting of NPC2 to lysosomes, and is crucial for NPC2 function. PMID- 15542394 TI - Determination of oligosaccharides and glycolipids in amniotic fluid by electrospray ionisation tandem mass spectrometry: in utero indicators of lysosomal storage diseases. AB - Prenatal diagnosis is available for many lysosomal storage disorders (LSD) using chorionic villus samples or amniocytes. Such diagnoses can be problematical if sample transport and culture are required prior to analysis. The purpose of this study was to identify useful biochemical markers for the diagnosis of lysosomal storage disorders from amniotic fluid. Amniotic fluid samples from control (n=49) and LSD affected (n=36) pregnancies were analysed for the protein markers LAMP-1 and saposin C by ELISA, and for oligosaccharide and lipid metabolite markers by electrospray ionisation-tandem mass spectrometry. Lysosomal storage disorder samples include; aspartylglucosaminuria, galactosialidosis, Gaucher disease, GM1 gangliosidosis, mucopolysaccharidosis types I, II, IIIC, IVA, VI, and VII, mucolipidosis type II, multiple sulfatase deficiency, and sialidosis type II. Each disorder produced a unique signature metabolic profile of protein, oligosaccharide, and glycolipid markers. Some metabolite elevations directly related to the disorder whilst others appeared unrelated to the primary defect. Many lysosomal storage disorders were clearly distinguishable from control populations by the second trimester and in one case in the first trimester. Samples from GM1 gangliosidosis and mucopolysaccharidosis type VII displayed a correlation between gestational age and amount of stored metabolite. These preliminary results provide proof of principal for the use of biomarkers contained in amniotic fluid as clinical tests for some of the more frequent lysosomal storage disorders causal for hydrops fetalis. PMID- 15542395 TI - Purification and characterization of recombinant murine sulfamidase. AB - Mucopolysaccharidosis type IIIA (MPS IIIA) is a lysosomal storage disorder caused by a deficiency in the lysosomal enzyme sulfamidase, which is required for the degradation of heparan sulfate. The disease is characterized by neurological dysfunction but relatively mild somatic manifestations. A naturally occurring mouse model to MPS IIIA exhibits a similar disease progression to that observed in patients. Disease in the mice results from a base substitution at codon 31 in the sulfamidase gene, altering an aspartic acid to an asparagine (D31N). This aspartic 31 is involved in binding of the divalent metal ion needed for catalytic function, and as such reduces the specific activity of the enzyme to about 3% of that of wild-type. The mutant protein has decreased stability and shows increased degradation over a 24 h chase period when compared to wild-type mouse sulfamidase. Mouse sulfamidase that was purified using a two-step ion exchange procedure was shown to have similar kinetic properties to that of purified human sulfamidase. Recombinant murine sulfamidase was able to correct the storage phenotype of MPS IIIA fibroblasts after endocytosis via the mannose-6-phosphate receptor. PMID- 15542396 TI - Expression and functional characterization of human mutant sulfamidase in insect cells. AB - Mucopolysaccharidosis IIIA (MPS IIIA; Sanfilippo syndrome) is an autosomal recessive lysosomal disorder caused by the deficiency of sulfamidase (EC 3.10.1.1), required for the degradation of the mucopolysaccharide heparan sulfate. The molecular defects of 26 unrelated Spanish MPS IIIA patients were recently reported by our group. Here we describe the heterologous expression, using a baculovirus system, of the cDNAs corresponding to eight out of the 14 mutant alleles present in this patient group and the characterization of the corresponding mutant enzymes. In particular, we expressed the following alleles: p.S66W, p.R74H, p.Q85R, p.R206P, p.L386R, p.R433W, p.R433Q, and c.1079delC (previously named as c.1091delC), and the two variants of the polymorphism p.R456H. The expression of the mutant alleles and the characterization of the corresponding enzymes revealed that their activity was severely compromised. Only mutations p.S66W and p.R206P retained low levels of residual activity. However, Western blot analysis showed in all cases the presence of the expected two forms of the sulfamidase, the precursor and the mature proteins, indicating a normal processing of the mutant enzyme. PMID- 15542397 TI - The PEX Gene Screen: molecular diagnosis of peroxisome biogenesis disorders in the Zellweger syndrome spectrum. AB - Peroxisome biogenesis disorders in the Zellweger syndrome spectrum (PBD-ZSS) are caused by defects in at least 12 PEX genes required for normal organelle assembly. Clinical and biochemical features continue to be used reliably to assign patients to this general disease category. Identification of the precise genetic defect is important, however, to permit carrier testing and early prenatal diagnosis. Molecular analysis is likely to expand the clinical spectrum of PBD and may also provide data relevant to prognosis and future therapeutic intervention. However, the large number of genes involved has thus far impeded rapid mutation identification. In response, we developed the PEX Gene Screen, an algorithm for the systematic screening of exons in the six PEX genes most commonly defective in PBD-ZSS. We used PCR amplification of genomic DNA and sequencing to screen 91 unclassified PBD-ZSS patients for mutations in PEX1, PEX26, PEX6, PEX12, PEX10, and PEX2. A maximum of 14 reactions per patient identified pathological mutations in 79% and both mutant alleles in 54%. Twenty five novel mutations were identified overall. The proportion of patients with different PEX gene defects correlated with frequencies previously identified by complementation analysis. This systematic, hierarchical approach to mutation identification is therefore a valuable tool to identify rapidly the molecular etiology of suspected PBD-ZSS disorders. PMID- 15542398 TI - Screening of SNPs at 18 positional candidate genes, located within the GD-1 locus on chromosome 14q23-q32, for susceptibility to Graves' disease: a TDT study. AB - Graves' disease (GD) is a complex autoimmune thyroid disorder with a strong genetic component. Genome-wide screens resolved several susceptibility loci that contribute to the development of GD. One of the susceptibility loci (GD-1 locus) was mapped on chromosome 14q31. However, a susceptibility gene located within the GD-1 locus remains undefined. Here we screen eighteen single nucleotide polymorphisms (SNPs), each is situated at a corresponding positional candidate gene, located within the GD-1 susceptibility locus on chromosome 14q23-q32, for predisposition to GD using the transmission disequilibrium test in 126 simplex Russian families affected with GD. Among SNPs tested, a significant preferential transmission of the Ala allele (41 transmissions vs. 17 nontransmissions, corrected P=0.031) of the Thr92Ala SNP within the DIO2 gene, encoding type II iodothyronine deiodinase, from parents to affected children was found in a Russian family data set. The Thr92Ala SNP of the DIO2 gene and the D727E substitution of the thyrotropin receptor (TSHR) gene have been found to be in pair-wise linkage disequilibrium. The A92/E727 haplotype showed significant preferential transmission from parents to affected sibling (17 transmissions vs. 8 nontransmissions, P=0.039) in simplex families. This suggests that the Thr92Ala variant of the DIO2 gene is associated or may be in linkage disequilibrium with a functional DIO2 polymorphism which involves in the development of GD in a Russian population. PMID- 15542399 TI - DNA-based subtyping of glycogen storage disease type III: mutation and haplotype analysis of the AGL gene in Chinese. AB - Glycogen storage disease type III (GSD III) is an inborn error of glycogen metabolism caused by a deficiency of glycogen debranching enzyme (AGL). Here, we investigate two unrelated Hong Kong Chinese GSD III patients and identify a novel 5-base pair deletional mutation, 2715_2719delTCAGAin exon 22, in one patient and a nonsense mutation, 1222C>T (R408X) in exon 11, in another patient. Since GSD IIIb is only caused by mutation in exon 3 of the AGL gene, we diagnose our patients to have GSD IIIa, which is consistent with the clinical diagnosis. Until now, R408X has only been reported in Faroe Islands GSDIII patients and was thought to demonstrate a founder effect. In this study, haplotyping of the disease-bearing chromosomes in the AGL locus by 19 intragenic single nucleotide polymorphisms shows that R408X is linked with IVS16+8T and IVS23-21T in our patient while R408X is linked with IVS16+8C and IVS23-21A in the Faroe Islands. The different haplotypes of R408X in Chinese and Faroese indicated that R408X is a recurrent mutation. PMID- 15542400 TI - Glycogen storage disease type Ia in Argentina: two novel glucose-6-phosphatase mutations affecting protein stability. AB - Glycogen storage disease type Ia (GSD-Ia) is caused by deleterious mutations in the glucose-6-phosphatase gene (G6PC). A molecular study of this gene was carried out in 11 Argentinean patients from 8 unrelated families. Four missense (p.Gln54Pro, p.Arg83Cys, p.Thr16Arg, and p.Tyr209Cys) and one deletion (c.79delC) mutations have been identified. Two novel mutations, p.Thr16Arg (c.47C>G) located within the amino-terminal domain and p.Tyr209Cys (c.626A>G) situated in the sixth transmembrane helix, were uncovered in this study. Site-directed mutagenesis and transient expression assays demonstrated that both p.Thr16Arg and p.Tyr209Cys mutations abolished enzymatic activity as well as reduced G6Pase stability. PMID- 15542401 TI - A single nucleotide polymorphism -1131T>C in the apolipoprotein A5 gene is associated with an increased risk of coronary artery disease and alters triglyceride metabolism in Chinese. AB - The disorder of triglyceride (TG) metabolism leading to hypertriglyceridemia is an independent risk factor for coronary artery disease (CAD). Variants in the newly identified apolipoprotein APOA5 gene were found to be strongly associated with elevated TG levels in different racial groups. In this study, we investigated the phenotypic effects of two polymorphisms (APOA5-1131T>C and APOC3 482C>T) on susceptibility to CAD in 312 Chinese CAD patients diagnosed by angiography. The frequency of the APOA5-1131C allele in these patients was significantly higher than that of the control group (39.9 vs. 33.3%, P=0.02). Compared with the wild type TT, CC homozygotes had a significantly increased CAD risk (OR=1.93 and OR=1.80 using unadjusted and adjusted logistic regression models, respectively). This association still existed after adjustment for the APOC3-482 variant. The APOA5-1131C allele also showed a correlation with increasing plasma TG levels (P<0.001). These data suggest that the APOA5-1131T>C polymorphism might contribute to an increased risk of CAD among Chinese as a result of its effect on TG metabolism; this effect was found to be independent of the APOC3-482C>T variant. PMID- 15542402 TI - Acute coronary syndromes presenting solely with heart failure symptoms: are they under recognised? PMID- 15542403 TI - The 6 minute walking test in chronic heart failure: indications, interpretation and limitations from a review of the literature. PMID- 15542404 TI - Differential expression of cytokines in the rat heart in response to sustained volume overload. AB - OBJECTIVE: The present study aimed to investigate whether sustained volume overload is capable of inducing persistent upregulation of cardiac cytokines including tumor necrosis factor alpha (TNF)-alpha, interleukin (IL)-1beta, interleukin (IL)-6 and transforming growth factor (TGF)-beta(1). METHODS AND RESULTS: Volume overload-induced heart hypertrophy in rats was established by aortacaval fistula, and the cardiac cytokines were measured in the myocardium from 1 to 4 weeks after operation. In the post-fistula rats, cardiac IL-1beta and IL-6 gene and protein levels were upregulated throughout the time of measurement. Immunohistochemistry demonstrated that IL-1beta and IL-6 immunoreactive cells were widely distributed in the myocardium in the earlier time intervals, and mainly localized in the regions close to the endocardium in the later time intervals. The cardiac IL-1beta immunoreactive cells were mainly localized in the blood vessels whereas the IL-6 positive cells were composed of non-myocytes and cardiomyocytes. TGF-beta(1) positive staining was increased in the myocardium up to 3 weeks after aortacaval fistula and then decreased to basal levels thereafter. In contrast to the activation of cardiac IL-1beta and IL-6 in response to volume overload, TNF-alpha expression appeared unaltered in response to sustained volume overload in the transcription and protein levels. CONCLUSION: The results of the present study indicate that sustained volume overload is capable of inducing persistent upregulation of some cardiac cytokines. In addition, the differential expressions of TNF-alpha, IL-1beta and IL-6 suggest that the induction of IL-6 and IL-1beta is independent of TNF-alpha mediated pathways in this animal model. PMID- 15542405 TI - Nitric oxide synthase inhibition impairs myocardial efficiency and ventriculo arterial matching in acute ischemic heart failure. AB - BACKGROUND AND AIMS: The effect of nitric oxide (NO) manipulation in acute heart failure has not been sufficiently investigated. Therefore, we assessed the impact of NO-synthase (NOS) inhibition on left ventricular (LV) function and energetics as well as overall hemodynamics, in a porcine model of acute ischemic LV failure. METHODS: Acute heart failure was induced by left coronary artery microembolization in fourteen anesthetized pigs. LV pressure-volume relationships and mechanical work (PVA) were assessed 30 min after stable heart failure, using pressure-conductance catheters. Myocardial oxygen consumption (MVO(2)) was determined from coronary flow and coronary arteriovenous oxygen difference. Microembolization led to a significant decrease in cardiac output, arterial pressure and LV systolic and diastolic performance. Animals were then randomized to a control group (n=7) or to receive 15 mg/kg N(omega)-Nitro-L-arginine-metyl ester (n=7), an inhibitor of NO synthase (NOS). RESULTS: Measurements 15 min later revealed that NOS inhibited animals had significantly reduced cardiac output (1.53+/-0.45 vs. 2.13+/-0.49 l/min, P=0.003) and stroke work (1054+/-461 vs. 1296+/-348 mmHg ml, P=0.03), and also displayed a significant increase in the slope of the MVO(2)-PVA relationship (2.57+/-0.53 vs. 1.92+/-0.15, P=0.008), i.e. an inefficient chemomechanical coupling. NOS inhibition did not alter contractility, diastolic function or arterial pressure, but afterload was significantly increased compared to controls (arterial elastance 6.03+/-1.48 vs. 2.74+/-0.34 mmHg/ml, P=0.009). CONCLUSION: Inhibition of NOS in experimental acute heart failure increased afterload without altering left ventricular systolic and diastolic function. Consequently, cardiac output was reduced. Furthermore, mechanoenergetic efficiency was severely impaired. NOS inhibition in acute heart failure and cardiogenic shock warrants further investigations. PMID- 15542406 TI - Cardiac rotation and relaxation in patients with chronic heart failure. AB - BACKGROUND: The normal left ventricle shows a systolic wringing motion with clockwise rotation at the base and counterclockwise rotation at the apex. PURPOSE: The aim of the present study was (1) to assess left ventricular (LV) contraction and relaxation in patients with chronic heart failure (CHF), and (2) to evaluate the effect of medical therapy on LV contraction-relaxation behavior. METHODS: Magnetic resonance was used to examine LV motion by labeling specific LV regions in three planes (myocardial tagging). Twenty-three subjects were included, nine healthy controls and 14 CHF patients. Cardiac motion was determined from the deformation of a rectangular grid in a basal and apical plane. CHF patients were put on triple therapy with ACE-inhibitors, beta-blockers and spironolactone. Follow-up examination (n=9) was performed after 6 months. RESULTS: In controls, systolic rotation was -9.5+/-2 degrees at the base and +3.3+/-1 degrees at the apex. In CHF patients, rotation was reduced both at the base (-3.4+/-2 degrees , P<0.01) and the apex (+0.9+/-3 degrees , P<0.05). Similarly, regional ejection fraction (REF) was reduced in CHF patients both at the base and the apex. Medical therapy was associated with an improvement in REF, but systolic rotation improved only at the base (-4.6+/-2 degrees , P<0.05). CONCLUSIONS: Systolic wringing motion with clockwise rotation at the base and counterclockwise rotation at the apex is maintained in CHF although reduced. Heart failure treatment is associated with an improvement in REF, whereas rotation improved only at the base, but not at the apex. Thus, there is an uncoupling between regional shortening and rotation in CHF patients. PMID- 15542407 TI - Clinico-pathological evaluation of restrictive cardiomyopathy (endomyocardial fibrosis and idiopathic restrictive cardiomyopathy) in India. AB - BACKGROUND: Restrictive heart disease is characterized by impairment of ventricular filling during diastole with preserved systolic function. The clinical and histopathological profile on endomyocardial biopsy of a cohort of patients with restrictive cardiomyopathy (RCM) is presented. METHODOLOGY: The medical records of patients presenting with heart failure with systemic congestion, subsequently diagnosed as restrictive heart disease after evaluation including cardiac catheterisation, were studied retrospectively to determine the clinical spectrum of restrictive cardiomyopathy. The diagnosis of RCM was made, based on systemic congestion with dilated atria and near normal ventricular size and function. Only patients who had an endomyocardial biopsy were included in the study. Patients with chronic constrictive pericarditis and secondary restrictive heart disease mainly amyloidosis were excluded from the study. RESULTS: All 52 patients had heart failure with normal or near normal left ventricular size and function. Based on right and left ventricle angiography, patients were classified into two groups. Group I with findings suggestive of EMF (n=30) and Group II no evidence of EMF on angiography i.e. 'idiopathic RCM' (IRCM) (n=22). Baseline characteristics were similar in the two groups. Echocardiography revealed typical features of endomyocardial fibrosis in Group I patients, with apical obliteration of right and left ventricular apices. Group II patients had no apex obliteration (except in four patients, who were misclassified and in whom angiography did not show apex obliteration). The Group II patients had features of IRCM in the form of normal left and right ventricular size and function with restrictive features of doppler filling along with dilated left and right atria. Angiocardiography in EMF patients showed isolated RV involvement in only two patients. In the remaining 28 patients, the obliterative changes were biventricular with RV involvement more severe than LV involvement. Angiographic findings in Group II (IRCM) patients were unremarkable with preservation of normal trabecular pattern and absence of obliterative changes. Mild atrioventricular regurgitation was present in 10/22 patients. Histopathological examination revealed that endocardial thickening was more common (77% vs. 23%) in EMF patients. The presence of myocyte hypertrophy (70-80%), myocytolysis (40-50%) and interstitial fibrosis (46-56%) were similar in both groups. CONCLUSIONS: The majority of our patients had biventricular EMF. A significant number of patients had clinical hemodynamic features of restrictive heart disease but no evidence of EMF on angiography. These IRCM patients had similar clinical profiles to EMF but on endomyocardial biopsy the endocardial thickening was minimal and seen in few patients (5/22). PMID- 15542408 TI - C-reactive protein predicts response to pentoxifylline in patients with idiopathic dilated cardiomyopathy. PMID- 15542409 TI - Restrictive filling pattern is associated with increased humoral activation and impaired exercise capacity in dilated cardiomyopathy. AB - OBJECTIVES: Although heart failure (HF) is characterized by increased proinflammatory cytokines, natriuretic peptide levels and impaired exercise capacity, the effect of concomitant diastolic dysfunction on those parameters has not been adequately studied. METHODS: We analyzed circulating levels of IL-1, IL 6, TNF-alpha and its soluble receptors, sTNFRI and sTNFRII, Nt-ANP and Nt-BNP natriuretic peptides in 81 patients, aged 56+/-12 years, with non-ischemic dilated cardiomyopathy (NIDC), LVEF 29.7+/-7.75% and functional NYHA class II III. An echocardiographic study and cardiopulmonary exercise test (CPE) were performed in all patients. RESULTS: Patients were divided into restrictive (24 patients, group I) and non-restrictive (57 patients, group II) groups, according to their transmitral-filling pattern. No differences in LV dimensions or LVEF were found between the two groups. Group I showed increased levels of IL-6 (P=0.006), TNF-alpha (P=0.05), sTNFRII (P=0.02), Nt-ANP (P<0.001) and Nt-BNP (P<0.001) and decreased exercise duration (P<0.001) and PVO(2) (P<0.001) compared to group II. The strongest independent predictors for restrictive filling pattern were Nt-ANP and IL-6 levels, while Nt-BNP levels were the strongest PVO(2) predictor. CONCLUSIONS: Restrictive filling pattern implying greater diastolic dysfunction may contribute to increased cytokine production in the heart failure syndrome, as well as greater increases in natriuretic peptides and decreased exercise tolerance. PMID- 15542410 TI - The effect of aspirin on the ventilatory response to exercise in chronic heart failure. AB - INTRODUCTION: Patients with chronic heart failure (CHF) experience breathlessness and fatigue on exercise. One of the abnormalities seen on maximal exercise testing is an increased ventilatory response to exercise (VE/VCO(2) slope). The cause of this is unknown, but is likely to be due to a combination of interacting peripheral and central factors. Recent data have demonstrated a relation between VE/VCO(2) slope and prostaglandin levels in contracting muscles. The present study examined the influence of the presence of a potent non-selective prostaglandin inhibitor, aspirin, on the ventilatory response to exercise in a group of patients with CHF. METHODS: We investigated the ventilatory response to exercise of 120 consecutive patients in sinus rhythm attending a specialist heart failure clinic. We excluded those taking clopidogrel (six patients) and those on both warfarin and aspirin or taking other non-steroidal anti-inflammatory agents (five patients). The other 109 patients were grouped according to whether they were taking aspirin (n=52 (48%)) or not (n=57 (52%)). Each patient underwent echocardiography to assess left ventricular function, and exercise testing with metabolic gas exchange to derive peak oxygen consumption (pVO(2)) and the VE/VCO(2) slope. RESULTS: The groups were similar in terms of age, (67 (13) vs. 66 (12) years; P=0.34) drug use, heart failure aetiology, left ventricular function (ejection fraction; 33.3 (9.4) vs. 31.8 (9.9)%; P=0.05)) and exercise tolerance (pVO(2); 20.4 (5.3) vs. 19.9 (6.0); P=0.68, and VE/VCO(2) slope; 35.4 (6.2) vs. 35.7 (9.3); P=0.73). There was no difference in the ventilatory response to exercise or the symptoms of breathlessness between the two groups. CONCLUSIONS: Aspirin does not appear to affect exercise performance in CHF. PMID- 15542411 TI - NT-proBNP: a marker for successful myocardial reperfusion in AMI patients treated with primary percutaneous coronary intervention. PMID- 15542412 TI - NT-brain natriuretic peptide levels in pleural fluid distinguish between pleural transudates and exudates. AB - BACKGROUND: Pleural effusion is not pathognomic and distinguishing between transudates and exudates often presents a diagnostic dilemma. The purpose of our study was to examine whether the inclusion of pleural fluid brain natriuretic peptide (BNP) measurement into the analysis improves the diagnostic accuracy of pleural effusion. METHODS: The pleural effusion of 14 patients with CHF (group A) and 14 subjects with different pleural pathology (group B) were analyzed. Samples of pleural fluid and serum were obtained from all patients on admission and biochemical analysis, bacterial and fungal culture, acid-fast bacilli smear and culture and cytology were performed on the pleural fluid. In vitro quantitative determination of N-terminal pro-Brain natriuretic peptide (NT-proBNP) in serum and pleural fluid were performed by electrochemiluminescence immunoassay proBNP method on an Elecsys 2010 (Roche) analyzer. RESULTS: The median NT-proBNP levels in groups A and B were 6295 pg/ml and 276 pg/ml, respectively: (P=0.0001). There was no overlap between the two groups. While the Light's criteria had a sensitivity of 93% and specificity of 43% for transudates, the pleural fluid NT proBNP level accurately differentiated between the two groups. CONCLUSIONS: The pleural NT-proBNP levels were elevated in all patients who had transudate. Therefore if the NT-proBNP levels of pleural effusion are within the normal range, transudate resulting from congestive heart failure can be ruled out. Our results suggest that the inclusion of pleural fluid NT-proBNP measurement in the routine diagnostic panel would enhance discrimination among the different causes of pleural effusions. PMID- 15542413 TI - Plasma brain natriuretic peptide levels in patients with rheumatic heart disease. AB - BACKGROUND: Brain natriuretic peptide (BNP) is a cardiac hormone secreted from the ventricular myocardium as a response to ventricular volume expansion and pressure overload. Rheumatic heart disease (RHD) is still an important cause of heart failure in developing countries. AIMS: To measure BNP levels in patients with RHD and to determine whether BNP concentrations correlate with clinical and echocardiographic findings. METHODS: Eighty-eight patients with rheumatic valve disease and 24 age- and sex-matched healthy subjects were entered in the study. BNP was measured using the Triage B-Type Natriuretic Peptide test (Biosite Diagnostics, San Diego, CA). Transthoracic echocardiography was performed in all patients to assess the severity of the valve disease and for the measurement of pulmonary artery pressure. RESULTS: The plasma concentrations of BNP were significantly higher in patients with rheumatic heart disease than in control subjects (232+/-294 vs. 14+/-12 pg/ml, p<0.0001). The plasma BNP level was significantly higher in NYHA class III+IV than in class II (463+/-399 vs. 192+/ 243 pg/ml, p<0.0001) and in NYHA class II than in class I (192+/-243 vs. 112+/ 135 pg/ml, p<0.001). The independent determinants of higher BNP levels were NYHA functional class and systolic pulmonary artery pressure in multivariate analysis. CONCLUSION: We found increased plasma BNP levels in patients with rheumatic heart disease compared with healthy subjects. PMID- 15542414 TI - Betablockers in heart failure: Carvedilol Safety Assessment (CASA 2-trial). AB - BACKGROUND: Betablockers are a cornerstone in the treatment of patients with chronic heart failure (CHF). The purpose of the present study was to assess safety and tolerability of carvedilol in CHF-patients. METHODS: 66 general practitioners, who were supervised by a local cardiologist, enrolled 151 CHF patients. All patients were on standard therapy with ACE-inhibitors and diuretics. Carvedilol treatment was started with 3.125 mg twice daily and slowly uptitrated in 2-week intervals to 2x25 mg per day. Mean follow-up was 12 weeks. RESULTS: 145 of the 151 patients (96%) finished the study according to protocol, six patients were lost to follow-up (4%). 59 patients (41%) experienced minor and nine (6%) serious adverse events. 68 were under maximal therapy with 50 mg daily, 33 received 25 mg, and 15 12.5 mg. Overall tolerability was good and NYHA-class fell significantly from 2.2 to 1.8 (P<0.001). Mean heart rate decreased from 78 to 69 bpm (P<0.001), mean systolic blood pressure from 137 to 132 mmHg (P<0.001) and mean diastolic blood pressure from 80 to 76 mmHg (P<0.001). Quality of life significantly improved under carvedilol with a reduction in the Minnesota living with heart failure score from 1.28 to 0.88 (P<0.001). CONCLUSIONS: Carvedilol is well tolerated in CHF-patients treated by general practitioners. Serious adverse events and hospitalisations are rare. Thus, carvedilol is a safe drug in the treatment of CHF-patients and can be easily initiated and managed by the general practitioner. PMID- 15542415 TI - The 'real' woman with heart failure. Impact of sex on current in-hospital management of heart failure by cardiologists and internists. AB - AIM: To identify differences between sexes in the clinical profile, use of resources, management and outcome in a large population of 'real world' patients with heart failure (HF). METHODS: A prospective cross-sectional survey was conducted on 2127 consecutive patients (47% women) admitted with HF to 167 cardiology and 250 internal medicine departments between February 14 and 25, 2000. RESULTS: Women were older, had a higher prevalence of atrial fibrillation, and more frequently a hypertensive or valvular aetiology. Females were admitted more frequently in Medical than in Cardiology Departments. The rate of invasive and non-invasive procedures was lower in women than in men, slightly higher if managed by cardiologists. Women were less frequently prescribed ACE-inhibitors, amiodarone, and spironolactone, and more frequently prescribed digoxin. In hospital mortality was similar, without difference between health-care providers. A 6-month follow-up was performed in 56.4% of the cases in both setting, but less frequently in women. Event rates were similar with nearly half of patients re hospitalised at least once. CONCLUSION: The 'real' HF woman has generally a more severe disease; she is an old lady who is more frequently hospitalised in a medical unit, receives few diagnostic, and cardiovascular procedures and pharmacological therapy, has a relatively low probability of dying in hospital, but a high likelihood of requiring readmission. PMID- 15542416 TI - Sex-related bedside clinical variables associated with survival of older inpatients with heart failure. AB - BACKGROUND: Little is known about sex-related differences in factors affecting prognosis of heart failure (HF). We prospectively investigated the relationship between bedside clinical variables and survival of older females vs. males with HF. METHODS: Included were consecutive unselected inpatients, age >/=60 years, admitted for various acute conditions. HF was chronic and of diverse etiologies. Follow-up extended up to 5 years. All-cause mortality was registered and statistically analyzed for association with in-hospital clinical variables. RESULTS: Included were 162 females and 200 males. Survival rates were 52.4% and 59%, respectively, (P=0.1). Advanced age and renal dysfunction (RD) were associated with low survival in both sexes (P<0.03 and 0.02, P<0.001 and 0.01, respectively). An association with low survival was found with respect to; admission for pulmonary edema (P<0.02), using furosemide >/=80 mg/day (P<0.005) and severe HF [NYHA class III-IV (P<0.01)] in females, as well as for hypokalemia (P<0.03) and hypocalcemia (P<0.03) in males. On multivariate analysis RD (P<0.001), increasing age (P=0.008) and furosemide dosage >/=80 mg (P=0.02) were most significantly associated with low survival in females, while RD only was significantly associated with low survival in males (P=0.03). CONCLUSIONS: Several clinical variables, which affect prognosis in older HF patients are sex related and probably carry practical significance. PMID- 15542417 TI - Clinical trials update from the European Society of Cardiology: SENIORS, ACES, PROVE-IT, ACTION, and the HF-ACTION trial. AB - This article provides information and a commentary on landmark trials presented at the European Society of Cardiology Congress in August 2004, relevant to the pathophysiology, prevention or treatment of heart failure. The SENIORS trial suggests that nebivolol is well tolerated and effective in older patients with heart failure, even if left ventricular systolic function is not markedly depressed. However, patients aged >75 years appeared to gain less benefit. Further data on the effects of nebivolol on symptoms and quality of life are awaited. Two new trials of long-term antibiotic prophylaxis after myocardial infarction (ACES and PROVE-IT) showed no benefit. The ACTION trial showed no reduction in serious cardiovascular events with nifedipine GITS in patients with chronic stable angina, despite a substantial reduction in blood pressure. The HF ACTION trial announced that the first 700 patients of a projected 3000 had been randomised to either an exercise program or encouragement to exercise but without a formal program. The primary outcome measure is death or hospitalisation for any reason. PMID- 15542418 TI - Diagnosing heart failure in primary care: lessons from EPICA. PMID- 15542419 TI - The diagnosis of heart failure in primary care: value of symptoms and signs. AB - BACKGROUND: The value of symptoms and signs in the diagnosis of CHF has rarely been tested in large numbers of patients in the community. The aim of this study was to evaluate the importance of symptoms, signs, and past medical history in the diagnosis of CHF in primary care. METHODS: Data on a sample of Portuguese men and women attending 365 primary care centres for any condition other than the treatment of acute infection, metabolic conditions or pregnancy were collected. All subjects who scored three or more points in the sum of categories one and two of the Boston questionnaire (history and physical examination) and those being treated for heart failure with loop or thiazide diuretics were considered to have possible heart failure and referred for further assessment including a resting echocardiogram. The sensitivity, specificity, positive predictive value, negative predictive value and likelihood ratio (LR) for the diagnosis of heart failure were calculated. RESULTS: A total of 5434 subjects were identified, of whom 1058 fulfilled the criteria for further assessment; 551 subjects had cardiac dysfunction at rest, of which 35.5% were in NYHA class I and 4.9% in class IV. Prior use of digoxin (LR 24.9) and/or diuretics (LR 10.6), a history of coronary artery disease (LR 7.1) or of pulmonary oedema (LR 54.2), were associated with a greater likelihood of having heart failure. Amongst current symptoms, a history of paroxysmal nocturnal dyspnoea (LR 35.5), orthopnea (LR 39.1) and breathlessness when walking on the flat (LR 25.8) were associated with a diagnosis of heart failure. However, these symptoms were not frequent amongst patients with heart failure within this population (sensitivity <36%). Jugular pressure > 6 cm with hepatic enlargement, and oedema of the lower limbs (LR 130.3), a ventricular gallop (LR 30.0), a heart rate above 110 bpm (LR 26.7), and rales at pulmonary auscultation (LR 23.3) were all associated with a diagnosis of heart failure, but were infrequent findings in patients with heart failure (sensitivity <10%). CONCLUSIONS: Symptoms and signs, and clinical history had limited value in diagnosing heart failure when used alone. The signs and symptoms that best predicted a diagnosis of heart failure were those associated with more severe disease. If investigation is limited to patients with more definite symptoms and signs of heart failure, fewer than 50% of cases will be identified and a large number of patients with mild symptoms will be missed. PMID- 15542420 TI - Aetiology, comorbidity and drug therapy of chronic heart failure in the real world: the EPICA substudy. AB - BACKGROUND: Chronic heart failure (CHF) is common and is frequently managed by primary care physicians (PCPs). Despite the European Society of Cardiology (ESC) Guidelines, standard treatments for CHF are frequently underutilised, particularly in primary care. AIM: To evaluate current drug therapy for CHF in adults with HF diagnosed according to ESC guidelines in the context of the EPICA study. Aetiological features and therapy relevant comorbidities were also analysed. METHODS: EPICA was a community-based epidemiological study conducted in mainland Portugal. The study involved 365 primary care physicians, who evaluated 6300 primary care attendees aged over 25 years. CHF was diagnosed by clinical and echocardiography criteria according to ESC guidelines. RESULTS: Total of 551 cases of CHF were identified, with a mean age of 65+/-9 years. The estimated overall prevalence of CHF in the Portuguese population was 4.4%; 1.3% with and 1.7% without left ventricular systolic dysfunction (LVSD). There are 6,280,792 people aged >25 years in Portugal, which extrapolates to 261,400 cases of heart failure. About 80% of patients had a history of hypertension, 39% had a history of coronary artery disease and 15% had atrial fibrillation. Only 58% of patients were on angiotensin-converting enzyme (ACE) inhibitors and 7% on beta-blockers. The type of ventricular dysfunction, age and presence of renal failure had little effect on prescription rates. Diuretics were prescribed in 78%. Thiazides were used more frequently in those with preserved systolic function and frusemide in those with left ventricular systolic dysfunction. Digoxin was prescribed more often to patients with than without left ventricular systolic dysfunction (34% vs. 17%; p=0.02). Long-acting nitrates were prescribed to 20% and amiodarone to 8% of patients. CONCLUSION: The EPICA study, as in other studies in primary care in Europe, particularly the IMPROVEMENT study, suggests that greater efforts are required to improve training of primary care teams in the management of CHF. PMID- 15542421 TI - The value of the electrocardiogram and chest X-ray for confirming or refuting a suspected diagnosis of heart failure in the community. AB - BACKGROUND: There is a common assumption that a normal ECG or a normal heart size on chest X-ray virtually rules out a diagnosis of heart failure. AIMS: To assess the value of the electrocardiogram and chest X-ray in identifying patients with chronic heart failure in the community. METHODS AND RESULTS: This study was a secondary analysis of data prospectively collected at the time of patient's enrollment in the EPICA study, an epidemiological study of the prevalence of heart failure in Portugal. A total of 6300 subjects were clinically evaluated. Patients who presented with symptoms or signs of heart failure, and/or were receiving diuretics for chronic heart failure (CHF) had a chest X-ray, ECG, and echocardiogram. The diagnosis of heart failure was confirmed in 551 cases. Patients with right atrial enlargement, atrial flutter, atrial fibrillation, 2nd degree-Mobitz I atrioventricular block, 1st degree atrioventricular block, left bundle branch block, lung interstitial oedema, and bilateral pleural effusion were more likely to be diagnosed with heart failure. For the diagnosis of heart failure, in the Portuguese population aged over 25 years, an abnormal electrocardiogram had an estimated sensitivity of 81%, and negative predictive value of 75%; an abnormal chest X-ray had an estimated sensitivity of 57%, and negative predictive value of 83%. Twenty five percent of patients with CHF had a normal ECG or chest X-ray. CONCLUSION: Our results show that electrocardiographic and roentgenographic features are not sufficient to allow heart failure to be reliably predicted in the community and support the recommendation that all patients with suspected heart failure should undergo echocardiography. PMID- 15542422 TI - Evaluation of the performance and concordance of clinical questionnaires for the diagnosis of heart failure in primary care. AB - AIM: To validate and estimate the performance statistics and concordance of seven clinical questionnaires for the diagnosis of chronic heart failure (HF). METHODS: Community-based epidemiological survey of patients aged >25 years attending a random sample of primary health care centers in Portugal. Heart failure was identified according to the Guidelines of the European Society of Cardiology (ESC). RESULTS: A total of 5434 subjects evaluated by 365 investigators were eligible for analysis, 551 of which had criteria for heart failure. Overall, the questionnaires had high specificity, usually above 90%, but low sensitivity, providing an increase in the likelihood of heart failure from 4.3% pre-test to 25 35% post-test in most cases. The Goteborg questionnaire was the most balanced regarding sensitivity (84%) and specificity (81%) but this may reflect its use of prescription of digoxin or diuretics as diagnostic criteria for diagnosis. The Walma, Framingham and NHANES-I questionnaires performed similarly (Sensitivity: 63%, Specificity: 93%), while the Boston and the Gheorghiade questionnaires had a somewhat lower sensitivity (55%). Concordance was good between the Boston, Framingham, Gheorghiade, NHANES-I and Walma questionnaires. CONCLUSIONS: This study evaluated seven clinical questionnaires for the diagnosis of heart failure in the community. Their low sensitivity impairs their usefulness as diagnostic instruments, but their high specificity makes them useful for the identification of patients with fluid retention and/or exercise intolerance from non-cardiac causes. PMID- 15542423 TI - HER2/HER3 heterodimers in prostate cancer: Whither HER1/EGFR? AB - In this issue of Cancer Cell, Mellinghoff et al. (2004) demonstrate that a small molecule inhibitor of the EGF receptor (EGFR) and the HER2/ErbB2/c-Neu kinase blocks signaling to the androgen receptor by a mechanism that involves HER2/HER3 heterodimerization. Surprisingly, the EGFR is peripheral to this signaling mechanism. These results have implications for the design of targeted therapy for hormone-refractory prostate cancer. PMID- 15542424 TI - A new key in breast cancer metastasis. AB - Unlocking the mysteries of cell metastasis, a major cause of cancer mortality, is essential in the development of novel therapies. In this issue of Cancer Cell, Li et al. (2004) identify a link between HER2 and CXCR4, two receptors previously implicated in breast cancer progression and metastasis. HER2 enhances the expression of CXCR4 by stimulating CXCR4 translation and attenuating CXCR4 degradation. Importantly, coexpression of HER2 and CXCR4 occurs in approximately 22% of human breast tumors and correlates with poor survival of breast cancer patients. PMID- 15542425 TI - Kick it up a Notch: NOTCH1 activation in T-ALL. AB - While the human NOTCH1 gene initially was cloned as part of a translocation breakpoint in T cell acute lymphoblastic leukemia (T-ALL) tumors, this translocation is present in only a small percentage of T-ALL patients. A recent paper by Weng et al. (2004) demonstrates that novel types of activating mutations in the NOTCH1 gene occur in more than half of all T-ALL cases, implicating NOTCH1 as a major player in the etiology of T-ALL. PMID- 15542426 TI - Somatic alterations in the human cancer genome. AB - Most human malignancies are caused by somatic alterations within the cancer genome, leading to oncogene activation or tumor suppressor gene inactivation. The sequence of the human genome has enabled systematic approaches to identify cancer genome alterations, including point mutations, copy number increases and decreases, loss of allelic heterozygosity, and chromosome translocations. Systematic cancer genome analysis has recently led to the discovery of somatic mutations in the BRAF, PIK3CA, and EGFR genes, among others. With further development of targeted cancer therapies and improvement in genome analysis technology, genome-wide surveys of cancer will likely become tools for diagnosis as well as discovery. PMID- 15542427 TI - Focus on multiple myeloma. PMID- 15542428 TI - RE: A two-gene expression ratio predicts clinical outcome in breast cancer patients treated with tamoxifen. PMID- 15542429 TI - Ras-induced interleukin-8 expression plays a critical role in tumor growth and angiogenesis. AB - The role of Ras oncogenes in promoting cellular transformation is well established. However, the contribution of Ras signaling to interactions between tumor cells and their host environment remains poorly characterized. Here, we demonstrate that the inflammatory mediator interleukin-8 (CXCL-8/IL-8) is a transcriptional target of Ras signaling. Using a tumor xenograft model, we show that Ras-dependent CXCL-8 secretion is required for the initiation of tumor associated inflammation and neovascularization. Collectively, our data identify a novel mechanism by which the Ras oncogene can elicit a stromal response that fosters cancer progression. PMID- 15542430 TI - Upregulation of CXCR4 is essential for HER2-mediated tumor metastasis. AB - The receptor tyrosine kinase HER2 enhances tumor metastasis; however, its role in homing to metastatic organs is poorly understood. The chemokine receptor CXCR4 has recently been shown to mediate the movement of malignant cancer cells to specific organs. Here, we show that HER2 enhances the expression of CXCR4, which is required for HER2-mediated invasion in vitro and lung metastasis in vivo. HER2 also inhibits ligand-induced CXCR4 degradation. Finally, a significant correlation between HER2 and CXCR4 expression was observed in human breast tumor tissues, and CXCR4 expression correlated with a poor overall survival rate in patients with breast cancer. These results provide a plausible mechanism for HER2 mediated breast tumor metastasis and establish a functional link between HER2 and CXCR4 signaling pathways. PMID- 15542431 TI - Integrin beta4 signaling promotes tumor angiogenesis. AB - Mice carrying a targeted deletion of the signaling portion of the integrin beta4 subunit display drastically reduced angiogenesis in response to bFGF in the Matrigel plug assay and to hypoxia in the retinal neovascularization model. Molecular cytology indicates that alpha6beta4 signaling promotes branching of beta4+ medium- and small-size vessels into beta4- microvessels without exerting a direct effect on endothelial cell proliferation or survival. Signaling studies reveal that alpha6beta4 signaling induces endothelial cell migration and invasion by promoting nuclear translocation of P-ERK and NF-kappaB. Upon subcutaneous implantation of various cancer cells, the mutant mice develop smaller and significantly less vascularized tumors than wild-type controls. These results provide genetic evidence that alpha6beta4 signaling promotes the onset of the invasive phase of pathological angiogenesis and hence identify a novel target for antiangiogenic therapy. PMID- 15542432 TI - Loss of HIF-1alpha in endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis. AB - We deleted the hypoxia-responsive transcription factor HIF-1alpha in endothelial cells (EC) to determine its role during neovascularization. We found that loss of HIF-1alpha inhibits a number of important parameters of EC behavior during angiogenesis: these include proliferation, chemotaxis, extracellular matrix penetration, and wound healing. Most strikingly, loss of HIF-1alpha in EC results in a profound inhibition of blood vessel growth in solid tumors. These phenomena are all linked to a decreased level of VEGF expression and loss of autocrine response of VEGFR-2 in HIF-1alpha null EC. We thus show that a HIF-1alpha-driven, VEGF-mediated autocrine loop in EC is an essential component of solid tumor angiogenesis. PMID- 15542433 TI - An autocrine mechanism for constitutive Wnt pathway activation in human cancer cells. AB - Autocrine Wnt signaling in the mouse mammary tumor virus model was the first identified mechanism of canonical pathway activation in cancer. In search of this transformation mechanism in human cancer cells, we identified breast and ovarian tumor lines with upregulation of the uncomplexed transcriptionally active form of beta-catenin without mutations afflicting downstream components. Extracellular Wnt antagonists FRP1 and DKK1 caused a dramatic downregulation of beta-catenin levels in these tumor cells associated with alteration of biological properties and increased expression of epithelial differentiation markers. Colorectal carcinoma cells with knockout of the mutant beta-catenin allele retained upregulated beta-catenin levels, which also could be inhibited by these Wnt antagonists. Together, these findings establish the involvement of autocrine Wnt signaling in human cancer cells. PMID- 15542434 TI - Suppression of angiogenesis and tumor growth by selective inhibition of angiopoietin-2. AB - Angiopoietin-2 (Ang2) exhibits broad expression in the remodeling vasculature of human tumors but very limited expression in normal tissues, making it an attractive candidate target for antiangiogenic cancer therapy. To investigate the functional consequences of blocking Ang2 activity, we generated antibodies and peptide-Fc fusion proteins that potently and selectively neutralize the interaction between Ang2 and its receptor, Tie2. Systemic treatment of tumor bearing mice with these Ang2-blocking agents resulted in tumor stasis, followed by elimination of all measurable tumor in a subset of animals. These effects were accompanied by reduced endothelial cell proliferation, consistent with an antiangiogenic therapeutic mechanism. Anti-Ang2 therapy also prevented VEGF stimulated neovascularization in a rat corneal model of angiogenesis. These results imply that specific Ang2 inhibition may represent an effective antiangiogenic strategy for treating patients with solid tumors. PMID- 15542435 TI - HER2/neu kinase-dependent modulation of androgen receptor function through effects on DNA binding and stability. AB - Given the role of the EGFR/HER2 family of tyrosine kinases in breast cancer, we dissected the molecular basis of EGFR/HER2 kinase signaling in prostate cancer. Using the small molecule dual EGFR/HER2 inhibitor PKI-166, we show that the biologic effects of EGFR/HER-2 pathway inhibition are caused by reduced AR transcriptional activity. Additional genetic and pharmacologic experiments show that this modulation of AR function is mediated by the HER2/ERBB3 pathway, not by EGFR. This HER2/ERBB3 signal stabilizes AR protein levels and optimizes binding of AR to promoter/enhancer regions of androgen-regulated genes. Surprisingly, the downstream signaling pathway responsible for these effects appears to involve kinases other than Akt. These data suggest that the HER2/ERBB3 pathway is a critical target in hormone-refractory prostate cancer. PMID- 15542440 TI - Hematopoietic secretory granules as vehicles for the local delivery of cytokines and soluble cytokine receptors at sites of inflammation. AB - Cytokines play an important role in the regulation of homeostasis and inflammation. A de-regulated cytokine function can subsequently promote chronic inflammation. This is supported by clinical evidence showing the beneficial effect of inhibiting TNF-alpha through injection of antibodies and soluble receptor in disorders such as rheumatoid arthritis and Crohn's disease. Systemic anti-TNF-alpha therapy however is associated with infectious complications. We therefore suggest a concept for the local deposition of therapeutically active agents into areas of inflammation or malignancy, based on the use of hematopoietic storage and secretory granules as delivery vehicles. Hematopoietic cells are induced to express the therapeutically active protein and to store it in the secretory lysosomes. The cells migrate into a tumour or site of inflammation, where the cells become activated and release the contents of their secretory lysosomes resulting in the local delivery of the therapeutically active protein. In support of this concept, gene transfer and granule loading can be achieved using the soluble TNF-alpha receptor (sTNFR1) after cDNA expression in hematopoietic cell lines. Endoplasmic reticulum (ER)-export can be facilitated by the addition of a transmembrane domain, and constitutive secretion can be prevented by incorporating a cytosol-sorting signal resulting in secretory lysosome targeting. The sTNFR1 is released from the transmembrane domain by proteolytic cleavage and finally, regulated sTNFR1-secretion can be triggered by a calcium signal. In vivo investigations are currently determining the feasibility of local protein delivery at sites of inflammation. PMID- 15542441 TI - Stromal cell-derived factor-1 (SDF-1)?CXCR4 couple plays multiple roles on haematopoietic progenitors at the border between the old cytokine and new chemokine worlds: survival, cell cycling and trafficking. AB - Generation of haematopoietic cells is regulated by cellular and humoral interactions in which stromal cells, adhesion molecules, cytokines and chemokines play a crucial role. Among the chemokines, SDF-1 and its CXCR4 receptor have been reported to be key players in the nesting of haematopoietic progenitors within the bone marrow. Disruption of the SDF-1?CXCR4 axis results in cell mobilization and may participate in leukaemia extramedullary infiltration. In this review we will discuss the manifold roles of the SDF-1 chemokine and of its receptor in haematopoiesis regulation. By recruiting quiescent progenitors, by participating in their survival?cycling and by sensitizing them to further cytokine synergistic action, SDF-1 likely contributes to haematopoiesis homeostasis under physiological conditions and in stress situations. The complexity of the SDF 1?CXCR4 interactions in the regulation of haematopoiesis illustrates a dynamic and sequential cross-talk between chemokine and cytokine?growth factor worlds. Because of their pleiotropic effects on haematopoietic progenitor trafficking, survival and proliferation, the SDF-1?CXCR4 couple could be considered as promising molecules for improvement of cell-based therapy protocols in haematopoietic transplantation. PMID- 15542442 TI - Plasmodium falciparum-specific interleukin-2 and tumor necrosis factor-alpha expressing-T cells are associated with resistance to reinfection and severe malaria in healthy African children. AB - The frequency of P. falciparum-specific interleukin (IL)-2-, interferon (IFN) gamma-, tumor necrosis factor (TNF)-alpha- and IL-10-expressing CD3+ cells was studied in healthy Gabonese children segregated according to their clinical presentation at admission to a longitudinal study of severe and mild malaria. The percentage of IL-2- and TNF-alpha- expressing P. falciparum-specific CD3+ cells was significantly higher in the children with prior mild malaria and less frequent reinfections compared to the children with prior severe malaria and more frequent reinfections. No differences were shown for P. falciparum-specific IFN gamma and IL-10 expression within CD3+ cells and parasite-non-specific expression of IL-2, IL-4, IL-6, IL-10, IL-13, TNF-alpha, and IFN-gamma within the CD4+, CD8+, TCRgamma?delta+ CD3+ and CD94+ CD3- cell populations, indicating that immunological determinants regulating the susceptibility to malaria in age matched children are parasite-specific. The ability of P. falciparum-specific T cells to mount a rapid IL-2 and TNF-alpha response might be of significance in preventing severe disease and reinfection. PMID- 15542443 TI - Short-term sequential analysis of sex hormones and helper T cells type 1 (Th1) and helper T cells type 2 (Th2) cytokines during and after multiple sclerosis relapse. AB - Multiple sclerosis (MS) is an immune-mediated disease with a clear sex-bias that may be attributed to sex hormones, sex' linked genes or both. Here we sought to determine the evolution pattern of cortisol and sex hormones at MS relapse and 2 months later in 7 male patients with relapsing remitting MS, and whether there was a correlation with a specific Th1 and Th2 cytokine pattern. Our findings indicate the activation of the hypothalamic-pituitary-adrenal axis and the concomitant upregulation of pro- and anti-inflammatory cytokines during relapse. The further increase of sex hormones, in particular estradiol in our male MS patients suggest their possible implication in the physiopathology of the illness and a putative anti-inflammatory and neuroreparatory effect. PMID- 15542444 TI - PADMA-28, a Tibetan herbal preparation is an inhibitor of inflammatory cytokine production. AB - BACKGROUND: Previous studies have shown that PADMA-28, a multicomponent, traditional Tibetan herbal plant preparation possesses a variety of beneficial effects on several experimental models of inflammatory and immune processes, including autoimmune diabetes and autoimmune encephalomyelitis. In humans, PADMA 28 attenuated the symptoms associated with intermittent claudications in atherosclerotic patients. OBJECTIVE: To assess the effect of PADMA 28 on the immune system, e.g. cytokine (interleukins) production. DESIGN: Cytokine production by human blood monocytes (derived from 12 healthy donors) stimulated in vitro, either by endotoxin (LPS) from Salmonella typhi or by lipoteichoic acid (LTA) from group A Streptococci was modulated by PADMA-28. RESULTS: The present study showed that an aqueous extract of PADMA-28 strongly decreased the production of the inflammatory cytokines IL-1beta, IL-6, IL-8 and TNF-alpha, and more moderately, also decreased the anti-inflammatory cytokine IL-10 induced by LPS. However, the LTA - induced IL-10 production was [not significantly] increased by the low dose PADMA-28, while not effected at all by the higher dose of PADMA-28. CONCLUSIONS: The data from these finding suggest a possible clinical efficacy of PADMA-28 either in autoimmune and in inflammatory conditions or in post-inflammatory sequelae, as previously shown in in vivo and human studies, probably by decreasing inflammatory cytokines. PMID- 15542445 TI - Elevated interleukin-18 protein expression in early active and progressive plaque type psoriatic lesions. AB - Psoriasis is a T cell-mediated inflammatory skin disease characterized by an elevated IFN-gamma and IL-12p70 expression in skin lesions. Interleukin-18 (IL 18) synergizes with IL-12 to induce IFN-gamma production and a strong T-helper-1 mediated immune response, or to induce Th2 polarization depending on the immunological context. We have previously shown that keratinocytes in normal skin produce and store large amounts of pro-IL-18. In this study, we hypothesized that the expression of IL-18 in psoriatic lesional skin might be altered compared to normal skin. Therefore, IL-18 expression was assessed in psoriatic, stable, plaque-type lesions and early active and progressive lesions. IL-18 mRNA and protein concentrations were constitutively high, and did not differ between normal and stable, plaque-type epidermis. In active and progressive lesions an elevated expression of total IL-18 protein relative to normal and stable, plaque type epidermis was detected using ELISA, while on Western blot, the differences in pro- or mature IL-18 were less clear. Our results indicate that the role of IL 18 in the pathogenesis of early phases of psoriasis may be more prominent than in established psoriatic lesions. PMID- 15542446 TI - Whole blood pro-inflammatory cytokines and adhesion molecules post lipopolysaccharides exposure in hyperbaric conditions. AB - Hyperbaric oxygen (HBO) is a therapeutic intervention with applications in a large variety of diseases, including traumatic injuries and acute or chronic infections. The presence of pro-inflammatory cytokines regulates certain factors including adhesion molecules, which play a significant role in HBO effects. We have investigated the effect of HBO on pro-inflammatory cytokine release [tumor necrosis factor-alpha (TNF-alpha), interleukin 6 and 8 (IL-6 and IL-8)], and the regulation of adhesion molecules [soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular adhesion molecule (sVCAM)] after lipopolysaccharide (LPS) stimulation in 16 healthy individuals, originating from an urban area. A total number of 64 samples were treated, divided into four groups: Group A: not stimulated with LPS and not exposed to HBO. Group B: stimulated with LPS and not exposed to HBO. Group C: not stimulated with LPS and exposed to HBO. Group D: stimulated with LPS and exposed to HBO. The LPS stimulation dose was 100 pg?ml for 0.1 ml whole blood diluted 1:10. After incubation, samples were exposed to HBO with 100% O2 at 2.4 atmospheres absolute (ATA) for 90 min. TNF-alpha, IL-6, IL-8 and sICAM-1, sVCAM levels were determined in culture supernatant, with ELISA. We observed an enhanced effect of LPS stimulation following exposure to HBO, which caused an increase in cytokine production (TNF-alpha, IL-6, IL-8), a reduction in sICAM, and no change to sVCAM, while their levels without stimulation remained almost invariable. The decrease in sICAM levels could be related to the increased levels of IL-8, as the production of this chemokine is involved in the regulation of adhesion molecules. PMID- 15542447 TI - Relationship between peripheral blood dendritic cells and cytokines involved in the pathogenesis of systemic lupus erythematosus. AB - Recent studies indicate that dendritic cells (DC) and several cytokines are implicated in the induction of autoimmune diseases. In this study we investigated the relationship between the total number of DC (tDC), and their plasmacytoid (pDC) and myeloid (mDC) subpopulations, with serum concentrations of interferons (IFN-alpha and IFN-gamma) and selected cytokines (TNF-alpha, IL-4, IL-6), in patients with systemic lupus erythematosus (SLE) and healthy persons. Subpopulations of DC were determined by the following antigen expression profiles: BDCA-1+?CD11c+?HLA-DR+ (for mDC) and BDCA-2 +?CD123+?HLA-DR+ (for pDC), using flow cytometry. Serum levels of interferons and cytokines were assessed by an enzyme-linked immunosorbent assay (ELISA). The study was performed in 36 SLE patients and 19 healthy volunteers. The mean number of tDC was lower in SLE patients (13.9 +/- 6.4?microL) than in healthy persons (24.1 +/- 12.6?microL) (P < 0.001). The number of pDC was also significantly lower in SLE (6.6 +/- 3.6?microL) than in the control group (12.0 +/- 8.3?microL) (P < 0.02). Moreover, the mean pDC count was lower in active than in inactive disease (5.5 +/- 3.6?microL vs 7.6 +/- 3.4?microL; P < 0.04). The mean serum levels of IFN-alpha and IFN-gamma were significantly higher in SLE patients (63.8 pg?mL and 6.6 pg?mL, respectively) than in the control group (2.7 pg?mL and 0.5 pg?mL, respectively) (P < 0.008 and P < 0.001, respectively). Serum levels of TNF-alpha and IL-6 were also higher in SLE patients (mean 7.3 pg?mL and 18.4 pg?mL, respectively) than in healthy controls (4.2 pg?mL and 0.5 pg?mL, respectively) (P < 0.02 and P < 0.001, respectively). The mean serum IL-4 concentrations were similar in SLE and healthy persons (0.2 pg?mL and 0.31 pg?mL, respectively; P -/+ 0.119). A negative correlation was found between pDC number and the serum level of IFN-alpha (rho -/+ -- 0.386, P -/+ 0.02) and between mDC and IFN-gamma (rho /+ -- 0.377, P -/+ 0.024). In conclusion, the correlation between peripheral blood DC subsets and serum levels of IFN-alpha and IFN-gamma suggests a possible relationship between these cytokines in the pathogenesis of SLE. PMID- 15542448 TI - Prolonged activation of tumor necrosis factor (TNF)-alpha and its soluble receptors in chronic heart failure patients both in the compensated and decompensated state. Interplay between their levels and metalloproteinase-3. AB - INTRODUCTION: Recent clinical and experimental studies indicate that upregulation of the TNF system can contribute to the progression of cardiac remodeling and heart failure decompensation, by promoting alterations in cardiomyocyte biology and extracellular matrix metabolism. Extracellular matrix turnover is regulated by the matrix metalloproteinases (MMPs), which are endogenous enzymes responsible for extracellular collagen degradation. The present study investigates the fluctuation of serum levels of TNF-alpha, soluble TNF receptor-1 (sTNFR1) and -2 (sTNFR2), in patients with chronic heart failure both during acute decompensation and the stable state of the syndrome. The second goal of this study was to determine if a relationship exists between serum MMPs profiles (MMP-1, MMP-2, MMP 3) and circulating TNF-alpha or its soluble receptors. METHODS: Our patient group consisted of 52 patients with chronic heart failure (NYHA III-IV; mean age: 65 +/ 4 years; hypertensive cardiomyopathy: 20, ischemic cardiomyopathy: 17, dilated cardiomyopathy: 10, valvular disease: 5), who were hospitalized for acute decompensation of the syndrome. Our control group consisted of 30 healthy subjects (mean age: 57 +/- 6 years). Serum levels of TNF-alpha, sTNFR1, sTNFR2 and MMP-1,-2,-3 were measured in heart failure patients by ELISA at admission and after one month as follow-up. Values are expressed as medians and interquartile ranges. RESULTS: In our patient group, we observed a statistically significant increase in the levels of sTNFR1 and sTNFR2 at admission (sTNFR1: 5.15 ng?mL, 4.49-8.90 ng?mL, P < 0.001, sTNFR2: 13.40 ng?mL, 6.10-21.50 ng?mL, P < 0.001), and at one-month follow-up (sTNFR1: 5.30 ng?mL, 4.61-6.90 ng?mL, P < 0.001, sTNFR2: 21.80 ng?mL, 11.50-25.20 ng?mL, P < 0.001), compared to the control group (sTNFR1: 3.83 ng?mL, 3.70-3.95 ng?mL, sTNFR2: 4.00 ng?mL, 3.40-5.40 ng?mL). There was a statistically significant difference in the levels of sTNFR2 between admission and follow-up (P < 0.05). Significant correlations between serum MMP-3 and sTNFR2 levels both at admission and follow up (r -/+ 0.460, P -/+ 0.005 and r -/+ 0.338, P -/+ 0.044, respectively) were also found. CONCLUSIONS: Soluble TNF receptors are elevated in heart failure patients both in acute decompensation and stable phase. We have detected higher levels of soluble TNFR2 during the compensated phase of heart failure, suggesting that TNFR2 receptors appear to stabilize the cytokine and thereby prolong its half-life and biological functions. Finally, TNF system-mediated cardiac remodeling may exist through the activation of MMP-3 signaling pathways. PMID- 15542449 TI - Expression of an interleukin-6 - interleukin-2 fusion protein (pIL-6-IL-2) in P. pastoris. AB - Interleukin-2 and interleukin-6 can stimulate the growth and proliferation of T lymphocytes and the differentiation of activated B lymphocytes respectively, and in turn enhance cellular and humoral immune responses. In this work, an expression clone using Pichia pastoris, a methylotrophic yeast strain, has been developed in order to produce large amounts of the functional recombinant fusion protein pIL-6-IL-2, which contains the mature porcine interleukin-6 peptide and the mature porcine interleukin-2 peptide. Two components of the fusion protein were connected by means of a flexible linker (Gly-Gly-Gly-Gly-Ser-Glu-Phe-Gly-Ser Gly-Gly). In response to 1% methanol induction, the recombinant strain GS115?9K IL6-IL2 secreted an exogenous protein, with a molecular weight of approximately 40 kD, into the culture medium. This was confirmed to be pIL-6-IL-2 by means of SDS-PAGE and Western Blot analysis. The protein was visible on the 2nd day following methanol induction, and peaked on the 4th day. By this time, the level had reached 50 mg?L as determined using the method of Bradford. After treatment with PNGase F and analysis of the concentration of sugar, the fusion protein pIL 6-IL-2 was further confirmed to be mainly a glycoprotein with an approximately 2 kDa sugar decoration. In addition, the IL-6 and IL-2 biological activities of the fusion protein, determined by cell proliferation assays using the IL6-dependent cell line B9 and the IL2-dependent cell line CTLL-2, reached 1 x 10(5) U?mg and 8 x 10(5) U?mg, respectively. This report is the first description of fused porcine cytokines expressed in P. pastoris, which might be an interesting adjuvant product for veterinary vaccines. PMID- 15542450 TI - Interleukine-17-induced inhibitory effect on late stage murine erythroid bone marrow progenitors. AB - Recent studies have shown that the T cell-derived cytokine, interleukin-17 (IL 17), stimulates hematopoiesis, specifically granulopoiesis inducing expansion of committed and immature progenitors in bone marrow. Our previous results pointed to its role in erythropoiesis too, demonstrating significant stimulation of BFU-E and suppression of CFU-E growth in the bone marrow from normal mice. As different sensitivities of erythroid and myeloid progenitor cells to nitric oxide (NO) were found, we considered the possibility that the observed effects of IL-17 were mediated by NO. The effects of recombinant mouse IL-17, NO donor (sodium nitroprusside - SNP) and two NO synthases inhibitors (L-NAME and aminoguanidine) on erythroid progenitor cells growth, as well as the ability of IL-17 to induce nitric oxide production in murine bone marrow cells, were examined. In addition, we tested whether the inhibition of CFU-E colony formation by IL-17 could be corrected by erythropoietin (Epo), the principal regulator of erythropoiesis. We demonstrated that IL-17 can stimulate low level production of NO in murine bone marrow cells. Exogenously added NO inhibited CFU-E colony formation, whereas both L-NAME and aminoguanidine reversed the CFU-E suppression by IL-17 in a dose dependent manner. The inhibition of CFU-E by IL-17 was also corrected by exposure to higher levels of Epo. The data obtained demonstrated that at least some of the IL-17 effects in bone marrow related to the inhibition of CFU-E, were mediated by NO generation. The fact that Epo also overcomes the inhibitory effect of IL-17 on CFU-E suggests the need for further research on their mutual relationship and co signalling. PMID- 15542451 TI - Expression of biologically active mouse ciliary neutrophic factor (CNTF) and soluble CNTFRalpha in Escherichia coli and characterization of their functional specificities. AB - Ciliary neurotrophic factor (CNTF) is a neuroprotective cytokine initially identified in chick embryo. It has been evaluated for the treatment of neurodegenerative diseases. CNTF also acts on non-neuronal cells such as oligodendrocytes, astrocytes, adipocytes and skeletal muscles cells. CNTF has regulatory effects on body weight and is currently in clinical trial for the treatment of diabetes and obesity. CNTF mediates its function by activating a tripartite receptor comprising the CNTF receptor alpha chain (CNTFRalpha), the leukemia inhibitory factor receptor beta chain (LIFRbeta) and gp130. Human, rat and chicken CNTF have been expressed as recombinant proteins, and most preclinical studies in murine models have been performed using rat recombinant protein. Rat and human CNTF differ in their fine specificities: in addition to CNTFR, rat CNTF has been shown to activate the LIFR (a heterodimer of LIFRbeta and gp130), whereas human CNTF can bind and activate a tripartite receptor comprising the IL-6 receptor alpha chain (IL-6Ralpha) and LIFR. To generate tools designed for mouse models of human diseases; we cloned and expressed in E. coli both mouse CNTF and the CNTFRalpha chain. Recombinant mouse CNTF was active and showed a high level of specificity for mouse CNTFR. It shares the arginine residue with rat CNTF which prevents binding to IL-6Ralpha. It did not activate the LIFR at all concentrations tested. Recombinant mouse CNTF is therefore specific for CNTFR and as such represents a useful tool with which to study CNTF in mouse models. It appears well suited for the comparative evaluation of CNTF and the two additional recently discovered CNTFR ligands, cardiotrophin-like cytokine?cytokine-like factor-1 and neuropoietin. PMID- 15542452 TI - Myeloid differentiation factor 88 (MyD88) is required for murine resistance to Candida albicans and is critically involved in Candida -induced production of cytokines. AB - We have studied the role of myeloid differentiation factor 88 (MyD88), the universal Toll-like receptor (TLR) adaptor protein, in murine defenses against Candida albicans. MyD88-deficient mice, experimentally infected in vivo, had a very significant impaired survival, and a higher tissue fungal burden when compared with control mice. The recruitment of neutrophils to the site of infection was also significantly diminished in MyD88-?- mice. In vitro production of proinflammatory cytokines such as TNF-alpha, IFN-gamma and IL-12p70, by antigen-stimulated splenocytes from mice intravenously infected with the low virulence C. albicans PCA2 strain, could not be detected in MyD88-?- mice. This default of production of Th1 cytokines in MyD88-deficient mice correlated with a greatly diminished frequency of IFN-gamma-producing CD4 + T lymphocytes. Also, the frequency of IFN-gamma-producing CD8 + T lymphocytes was lower in MyD88-?- mice than in control mice. Although C. albicans-specific antibody titers in PCA2 infected mice appeared more quickly in MyD88-?- mice than in control mice, the MyD88-?- group was not able to maintain the Candida-specific IgM nor IgG titers at the third week of infection. The complexity of antigens recognized by sera from MyD88-?- mice was quite similar to that from infected control mice. Taken together, these data show that MyD88-?- mice are extremely susceptible to C. albicans infections, suggesting that MyD88-dependent signaling pathways are essential for both the innate and adaptive immune responses to C. albicans. PMID- 15542454 TI - Reevaluating thyrotropin receptor-induced mouse models of graves' disease and ophthalmopathy. AB - We aimed to establish and extend the characterization of murine models of thyroiditis and Graves' ophthalmopathy, induced by transfer of TSH receptor (TSHR) primed T cells. Experiments were performed in a different animal unit but using female BALB/cbyJico mice from the same supplier as previously. We report our findings together with a reevaluation of the earlier studies. In the first experiment, genetic immunization or TSHR fusion protein induced TSHR antibodies in all nine mice. Some of the antibodies functioned as thyroid-stimulating antibodies and/or TSH binding inhibiting Igs with two of seven mice having elevated T4. Thyroiditis and orbital changes were absent. Splenocyte transfer induced no immune response in naive BALB/cbyJico recipients. Subsequently genetic immunization or fusion protein-treated mice were maintained in either local or Brussels conditions (water, chow, and bedding). TSHR antibodies were induced in nine of nine Brussels (with decreased T4 in one of nine) but five of nine local mice. No thyroiditis or orbital changes were induced, but misleading fixation artefacts in extraocular muscles were noted. Nonspecific in vitro stimulation induced more CD-4+/IL-4+ cells in Brussels maintained. TSHR stimulation produced a significant increase in IL-4 secretion in six of nine local but one of seven Brussels mice. Thyroids from many TSHR-treated and control mice contained ectopic thymus. Our results confirm that thyroiditis is required for disease transfer but indicate the heterogeneity in TSHR-induced immune response in an inbred strain. Ectopic thymus can masquerade as thyroiditis, and care is required to avoid muscle artefacts. Because neither animal unit is pathogen free, microbial environment may contribute to determining TSHR-induced responses. PMID- 15542455 TI - NIH Molecular Libraries Initiative. PMID- 15542456 TI - Genetics of length of productive life and lifetime prolificacy in the Finnish Landrace and Large White pig populations. AB - The objective of this study was to estimate direct and indirect selection potential for length of productive life and lifetime prolificacy in Finnish Large White and Landrace swine populations. To study the direct selection potential, the heritabilities of these traits were estimated. The genetic correlations of length of productive life and lifetime prolificacy with prolificacy traits and overall leg conformation were estimated to evaluate whether selection for these traits could indirectly improve measures of sow longevity. In addition, correlations between length of productive life, lifetime prolificacy, ADG, and backfat thickness were estimated. Records were used from Finnish purebred Landrace (n = 26,744) and Large White (n = 24,007) sows born on operations that perform on-farm production tests on all females. Heritabilities were estimated using both a survival analysis procedure and a linear model. Due to computational limitations, correlations were estimated with the linear model only. Estimated length of productive life heritabilities obtained from linear model analyses were less (0.05 to 0.10) than those obtained from survival analyses (0.16 to 0.19). This may be indicative of the superiority of survival analysis compared with linear model analysis methods when evaluating longevity or similar types of data. All the prolificacy traits were genetically correlated with length of productive life and lifetime prolificacy, and the correlations were greater than 0.13. These results indicate that selection for increased number of piglets weaned in the first litter and for short first farrowing interval is beneficial for sow longevity and also for sow's lifetime prolificacy. The genetic correlations between length of productive life and leg conformation score also were favorable (0.32 in Landrace and 0.17 in Large White). The heritability estimates indicate that survival analysis is likely the most appropriate method of evaluating longevity traits in swine. Because of computational problems, simultaneous analysis of linear traits and longevity is not currently possible. More research is needed to develop methods for multiple linear and survival trait analyses. PMID- 15542457 TI - Genetics of osteochondral disease and its relationship with meat quality and quantity, growth, and feed conversion traits in pigs. AB - The main objective of this research was to estimate heritabilities of seven osteochondrosis (OC) lesions in station-tested pigs and their genetic and phenotypic correlations with four meat quality (MQ) traits, the percentage of premium cuts (PPC), daily weight gain (DWG), and feed conversion ratio (FCR). Observed OC lesions were on the head of humerus (HK), condylus medialis humeri (CMH), condylus lateralis humeri (CLH), radius and ulna proximal (RUP), distal epiphyseal cartilage of ulna (DEU), head of femur (FK), and condylus medialis femoris (CMF). Meat quality traits were i.m. fat (IMF), muscle pH at 1 h after slaughter (pH1), muscle pH at 30 h after slaughter (pH30), and light reflectance on muscle (H30). The data set comprised 2,710 animals, of which 1,291 animals had OC records. All traits were analyzed by multiple-trait linear mixed model, with the animal's genetic and common litter effects as random. Fixed effects in the model varied between traits. Each OC lesion was further analyzed by a univariate generalized linear mixed model or, equivalently, "threshold models," assuming logistic, probit (normal), and Poisson distributions of the underlying "liability" to the disease. For OC lesions, estimates of heritability were low on the original "incidence" scale (0.06 for HK to 0.16 for CLH) and moderate to high on the liability scale (0.08 to 0.42). Genetic correlations (r(g)) between OC lesions and most MQ traits and PPC were generally unfavorable. Significant r(g) were -0.44 for DWG-CMH, 0.31 for DWG-CMF, 0.40 for FCR-HK, 0.21 for PPC-CLH, 0.32 for PPC-RUP, 0.30 for PPC-CMF, -0.54 for pH1-CLH, 0.47 for pH1-DEU, -0.34 for pH30-CMH, 0.58 for pH30-DEU, -0.50 for H30-HK, -0.31 for H30-DEU, and 0.31 for H30-CMF. Genetic susceptibilities to some OC lesions within the front leg were positively related to each other (r(g) range = 0.57 to 0.69), but r(g) between front and hind leg OC lesions were mostly negative (range = -0.21 to -0.40). Estimated h2 was 0.60 for PPC, and ranged from 0.12 to 0.66 for MQ traits, 0.28 for DWG, and 0.42 for FCR. Genetic correlations among meat quality and quantity traits ranged from -0.66 to 0.37. This is the first study to report genetic and phenotypic correlations between OC lesions and several meat quality and quantity traits in pigs. These findings will be useful to pig industry, especially in designing breeding programs for robust pigs. PMID- 15542458 TI - Effects of a quantitative trait locus for muscle hypertrophy from Belgian Texel sheep on carcass conformation and muscularity. AB - A QTL for muscle hypertrophy has been identified in the Belgian Texel breed. A population of F2 and backcross lambs created from crosses of Belgian Texel rams with Romanov ewes was studied. Effects on carcass traits and muscle development of the Belgian Texel breed polygenes and Belgian Texel single QTL were compared. In both cases, carcass conformation and muscularity were improved. The Texel polygenic environment improved conformation mainly through changes in skeletal frame shape. Segments were shorter and bone weight lower. Muscles were more compact, shorter, and thicker. The single QTL affected muscle development. Thickness and weight of muscles were increased. Composition in myosin changed toward an increase of fast contractile type. The relative contribution of hind limb joint to carcass weight was increased. Differences in skeletal frame morphology among the three genotypes of the single QTL were small. Conformation scoring was mainly influenced by leg muscularity. Back and shoulder muscle development, which largely contributed to variability of muscularity, were less involved in the conformation scoring. Lastly, the QTL explains a small part of differences between these Belgian Texel and Romanov breeds for conformation or muscle development. A large part of genetic variability remains to be explored. PMID- 15542459 TI - The effect of selection for growth rate and slaughter age on carcass composition and meat quality traits in rabbits. AB - The effect of selection for growth rate on carcass and meat quality was assessed by comparing selected and control populations of rabbits measured at the same stage of maturity and slaughtered at 9 and 13 wk of age. Embryos belonging to Generation 7 were frozen, thawed, and implanted in does to produce the control group. The control group was formed from the offspring of the embryos belonging to the Generation 7. Selected animals belonging to Generation 18 (S) were compared with contemporary animals of the control group (C). Carcasses were dissected and measured according to World Rabbit Science Association recommended practices. When animals were compared at similar degrees of maturity, selection for growth rate did not produce a negative effect on carcass and meat quality. There was no increase in fat content of the carcass, and there was an improvement of the meat:bone ratio with selection, with a difference between C and S groups of -0.42. However, the carcasses of S animals have 1.45% lower water-holding capacity. Carcass quality changed markedly with animal age. Heavy rabbit carcasses had lower organ percents and a higher loin percent. Dissectible and i.m. fat content were higher in older rabbits, with older animals having 0.97 and 0.79% more dissectible and i.m. fat content, respectively. Meat quality traits improved with age of slaughter, although there was an increase in glycolytic metabolism. Results from this study indicate that selection for growth rate has little effect in carcass and meat quality when rabbits are measured at the same stage of maturity. PMID- 15542460 TI - Evaluation of Dorset, Finnsheep, Romanov, Texel, and Montadale breeds of sheep: IV. Survival, growth, and carcass traits of F1 lambs. AB - The objectives of this study were to estimate effects of sire breed (Dorset, Finnsheep, Romanov, Texel, and Montadale), and dam breed (Composite III and Northwestern whiteface) on survival, growth, carcass, and composition traits of F1 lambs. Effects of mating season (August, October, and December) were estimated for survival and growth traits. Data were collected on 4,320 F1 lambs sired by 102 purebred rams over 3 yr. Birth weight was recorded on all lambs, and subsequent BW were adjusted to 56 (weaning), 70, and 140 d of age (n = 3,713, 3,654, and 3,579 observations, respectively). Survival of dam-reared progeny (n = 4,065) to weaning was recorded. Each year, wethers from October matings were slaughtered in three groups at 25, 29, and 33 wk of age to obtain carcass data (n = 546). In addition to standard carcass traits, resistive impedance measurements were recorded on the warm carcass to predict lean mass. Dam breed (P = 0.37) did not influence lamb survival to weaning, but sire breed (P < 0.05) was important. Romanov-sired lambs excelled in survival rate to weaning (94.1%), followed by Finn-sheep (93.0%), Texel (90.7%), Dorset (90.0%), and Montadale (89.1%) sired progeny. Lower (P < 0.01) postweaning growth rate was observed for Texel (267 g/d) and Finnsheep (272 g/d) sired progeny than for Dorset (285 g/d), Montadale (282 g/d), and Romanov (278 g/d) sired progeny. Sire breed and dam breed were generally significant for most carcass traits. Breed differences in distribution of carcass fat and carcass shape were detected; however, carcass composition was similar for all sire breeds when compared at a constant carcass weight. When evaluated at a constant 12th-rib fat depth, carcasses of lambs from Finnsheep, Romanov, and Texel sires produced 1 to 1.5 kg less (P < 0.001) predicted lean mass per lamb than carcasses of lambs from Dorset and Montadale sires. These experimental results provide information about the direct breed effects for survival, growth, and carcass traits of these breeds and their potential use in crossbreeding systems. PMID- 15542461 TI - Conceptus and maternal responses to increased feed intake during early gestation in pigs. AB - Maternal diet influences fetal growth and postnatal development. We hypothesized that conceptuses gestated in sows provided ad libitum vs. restricted feed intake would differ in the milieu of hormones, growth factors, nutrients, and metabolites associated with growth and metabolism. This hypothesis was tested in two experiments by providing fourth-parity sows (Pig Improvement Co. C15 bred to Line 326 boars) with either 1.81 kg/d (as-fed basis; control) or ad libitum access to gestation diet. In Exp. 1, control (n = 6) or ad libitum (6.4 +/- 0.11 kg/d; n = 9) treatments were provided from d 29 to 45 (onset of estrus is d 0), and sows were slaughtered on d 46. Ad libitum sows gained more weight from d 29 to 45 than controls (34.0 vs. 4.32 kg, respectively; P < 0.01). No differences were observed on d 46 for the number of fetuses, conceptus attachment length, allantoic + amniotic fluid volume, placental weight, fetal weight, and fetal crown-to-rump length. Variation in fetal crown-to-rump length was less (P < 0.03) in sows fed ad libitum. Sows fed ad libitum had greater (P < 0.01) IGF-I and insulin concentrations in plasma than controls on d 43. In Exp. 2, sows were fed 1.81 kg/d (n = 6) or ad libitum (7.0 +/- 0.11 kg/d; n = 4) from d 30 to 56 of gestation, when sows were anesthetized and samples were collected surgically from their gravid uteri. Sows fed ad libitum gained more weight (P < 0.01) than did controls and had more (P < 0.06) IGF-I in their plasma and the plasma collected from umbilical veins of their fetuses. No differences were found for concentrations of insulin or glucose in plasma of sows or fetuses, but urea N concentrations were greater (P < 0.05) in maternal plasma and in the plasma, and allantoic and amniotic fluids of conceptuses from sows fed ad libitum. Combined data from Exp. 1 and 2 revealed a treatment x fetal number interaction (P < 0.05) for average fetal weight. The expected negative relationship between within litter average fetal weight and the number of fetuses per uterus was observed for control sows (y = 115.4 -1.75 x fetal number; P < 0.05), but litters of ad libitum sows did not show this effect. The hypothesis that providing feed in excess of established requirements in early gestation affects the in utero milieu is supported by these results. Data further reveal that, at least at mid gestation, the restraint to fetal growth that is exhibited when fetal number increases in control sows is not exhibited when sows are fed ad libitum. PMID- 15542462 TI - Cloning and expression of porcine adiponectin and adiponectin receptor 1 and 2 genes in pigs. AB - In mice, adiponectin receptors (AdipoR) have been found to mediate the effect of adiponectin in muscle and liver in regulation of glucose and fatty acid metabolism. The purposes of this study were to clone these receptors from pig tissues by reverse transcription PCR using mRNA from skeletal muscle and adipose tissue and to investigate the expression of these genes in various pig tissues. Sequences of adiponectin, AdipoR1, and AdipoR2 were determined and found to be highly homologous to those of the human and mouse. The AA sequences predicted for the full-length cDNA of porcine adiponectin, AdipoR1, and AdipoR2 were similar to those of the human and mouse, ranging from 81 to 97% homology, suggesting similar functions of these genes in pigs as in other species. Transcripts for adiponectin were abundant in s.c. adipose tissue in Lee-Sung pigs and in crossbred pigs. Transcripts for AdipoR1 were abundant in heart and skeletal muscle and also detected to a lesser extent (P < 0.05) in adipose tissue, liver, and spleen of the Lee-Sung pigs. Transcripts for AdipoR2 were abundant in s.c. adipose tissue and present to a lesser extent (P < 0.05) in the liver, heart, skeletal muscle, and spleen. These results indicate that the effect of adiponectin may be mediated through these receptors in various porcine tissues. Fasting for 8 h did not have a significant effect on the expression of adiponectin and AdipoR1 mRNA, but it increased (P < 0.05) the AdipoR2 mRNA in the s.c. adipose tissue of crossbred pigs. These results indicate that the AdipoR2-mediated fatty acid oxidation may be responsible at least in part for the fasted state fatty acid oxidation in porcine adipose tissues. The successful cloning of pig adiponectin and adiponectin receptors will enhance the understanding of the involvement of these genes in regulating energy metabolism in pigs. PMID- 15542463 TI - Effects of elevated temperature in vivo on the maturational and developmental competence of porcine germinal vesicle stage oocytes. AB - Postslaughter processing of sow carcasses results in the ovaries being exposed to temperatures of 41.3 to 42.1 degrees C within a 30-min time frame. This study investigated whether the maturational and developmental competence of the recovered germinal vesicle stage oocytes could be compromised by post-slaughter processing. The results showed that the in vitro maturation rates of GV stage oocytes exposed to elevated temperature did not significantly differ from the corresponding controls (74.1 vs. 75.8%). Immunocytochemical staining revealed that elevated temperature did not adversely affect metaphase II spindle formation but resulted in extensive disruption of oocyte cytoskeletal organization. This, in turn, had a detrimental effect on parthenogenetic development compared with the corresponding nonheat-treated controls (cleavage rate = 27.7 vs. 65.3%, P < 0.01; blastulation rate = 6.7 vs. 20.6%, P < 0.01). Hence, transient exposure to elevated temperature during slaughter did not have any detrimental effects on nuclear maturation per se, but it did result in extensive cytoskeletal damage, which in turn drastically decreased the developmental competence. PMID- 15542464 TI - Abundance of mRNA encoding for components of the somatotropic axis and insulin receptor in different layers of the jejunum and ileum of neonatal calves. AB - Insulin-like growth factors-1 and -2, IGFBP-2 and -3, and receptors for IGF type 1 and type-2 (IGF-1R, IGF-2R), growth hormone (GHR), and insulin (InsR) in neonatal calves are variably expressed among gastrointestinal sites and thought to exert site-specific physiological functions. We studied by real-time reverse transcription PCR, whether there are differences in the abundance of mRNA coding for IGF-I, IGF-2, IGFBP-2, IGFBP-3, IGF-1R, IGF-2R, GHR, and InsR in compartmentalized layers (fractions) of jejunum and ileum of 5-d-old calves fed colostrum. Samples of jejunum consisted primarily of villi and crypts; samples from ileum consisted mainly of villus tips, crypts, and lamina propria (LP; containing mainly Peyer's patches). After slaughter, segments of middle areas of jejunum and ileum were flushed with 154 mM NaCl. Pieces (5 mm x 5 mm) of jejunal (n = 9) and ileal walls (n = 5) were placed on glass slides and snap-frozen in liquid N before being cut horizontally into 10-mum-deep slices using a cryotome at -20 degrees C. Fifteen consecutive and morphologically similar slices were collected as fractions of villus, crypt, and LP layers, respectively. Fractions were characterized by use of 5'-bromo-2-deoxyuridine (BrdU) that labeled proliferating cells, and by expression of lactase mRNA. The BrdU-labeled cells were present in crypts and LP, but not in tips of villi. Lactase mRNA levels were greater in villus than crypt fractions, but lactase mRNA was absent in LP. In jejunum, mRNA levels, relative to levels of housekeeping genes (sum of levels of mRNA coding for ubiquitin, glyceraldehyde phosphate dehydrogenase, beta-actin, and ribosomal RNA), differed (P < 0.05) between fractions for InsR (crypts > villi), IGFBP-2 (crypts > villi), and IGFBP-3 (crypts > villi), and total RNA levels were greater (P < 0.05) in crypt than villus fractions. In ileum, mRNA levels, expressed relative to housekeeping genes, differed (P < 0.05) between fractions for IGF-I (LP > villi, crypts), IGF-2, and IGFBP-3 (villi > crypts, LP), GHR and InsR (crypts > LP), IGFBP-2 (crypts > villi, LP), and total RNA levels were greater (P < 0.05) in LP and crypt than in villus fractions. In conclusion, the tested fractionation technique is quite applicable for gene expression studies in the intestine of calves. Members of the somatotropic axis and of the insulin receptor are not equally expressed in different jejunal and ileal layers of neonatal calves. PMID- 15542465 TI - Effects of dietary iron supplementation on growth performance, hematological status, and whole-body mineral concentrations of nursery pigs. AB - An experiment was conducted to evaluate the effects of supplementing increasing concentrations of Fe to the diet of nursery pigs on growth performance and indices of hematological and mineral status. Pigs (n = 225; 6.5 kg; 19 +/- 3 d) were allotted randomly by BW, litter, and gender to one of five dietary treatments (five pigs per pen; nine pens per treatment). Basal diets for each phase (Phase 1: d 0 to 7; Phase 2: d 7 to 21; Phase 3: d 21 to 35) were formulated to contain minimal Fe concentration and then supplemented with 0, 25, 50, 100, and 150 mg Fe/kg of diet (as-fed basis) from ferrous sulfate. Three pigs per pen (n = 135) were chosen and bled throughout (d 0, 7, 21, and 35) to determine hemoglobin (Hb), hematocrit (Hct), transferrin (Tf), and plasma Fe (PFe). In addition, pigs (n = 5; 5.9 kg; 19 +/- 3 d) from the contemporary group were killed at d 0 to establish baseline (BL), and 30 pigs (six pigs/treatment) were killed at d 35 to determine whole-body and liver mineral concentrations. The improvements in growth performance during Phase 2 (ADG = linear, P = 0.04; ADFI = linear, P = 0.10; G:F = quadratic, P = 0.07) were of sufficient magnitude that dietary treatments tended to increase ADG (linear, P = 0.08), ADFI (quadratic, P = 0.09), and G:F (quadratic, P = 0.10) for the 35-d experiment. Hematological variables were not affected until d 21, at which time dietary Fe supplementation resulted in a linear increase (P = 0.03) in Hb, Hct, and PFe. This linear increase (P = 0.001) was maintained until d 35 of the experiment; however, dietary treatments resulted in a linear decrease (P = 0.01) in Tf on d 35. Whole body Fe concentration increased (linear, P = 0.01) in pigs due to increasing dietary Fe concentrations. Moreover, pigs fed for 35 d had greater (P = 0.02) whole-body Fe, Zn, Mg, Mn, Ca, and P concentrations and lower (P = 0.001) whole body Cu concentration than BL. Hepatic Fe concentration increased (linear, P = 0.001) in pigs due to dietary treatments; however, the hepatic Fe concentration of all pigs killed on d 35 was lower (P = 0.001) than the BL. Results suggest that Fe contributed by feed ingredients was not sufficient to maintain indices of Fe status. The decrease in Fe stores of the pigs was not severe enough to reduce growth performance. Even so, the lessening of a pig's Fe stores during this rapid growth period may result in the occurrence of anemia during the subsequent grower and finisher periods. PMID- 15542466 TI - Ileal amino acid digestibilities by pigs fed soybean meals from five major soybean-producing countries. AB - Growing conditions and processing technologies to which soybeans (SB) are exposed have an effect on digestibilities of AA found in the resultant soybean meals (SBM). This study evaluated SBM from five major SB-producing countries (Argentina, Brazil, China, India, and the United States). An industry representative in each country collected samples of unprocessed SB and SBM subjectively determined to be of high, intermediate, or low quality. The SB from each country were processed into SBM under uniform conditions in the United States. Five experiments (each examining the three SBM and the SB processed in the United States from a single country) were conducted to determine true ileal AAd digestibilities. In addition, a standard SBM purchased on the open market in the United States was used in all experiments as a control. Data from pigs fed a low-protein casein diet in each study were used to calculate true AA digestibilities. Pigs were fitted with simple T-cannulas at the terminal ileum and allotted to treatments in Latin square design experiments. Duplicate experiments were conducted at the University of Illinois and at The Ohio State University. Within each country comparison, pigs fed the SBM processed in the U.S. from SB grown in the five countries had lower (P < 0.05) true total amino acid (TAA) digestibilities than did pigs fed any of the SBM prepared within the country of origin, except the United States. This indicates that processing conditions used at the U.S. pilot plant were not ideal when using SB from other countries. True TAA digestibilities of the diets containing the high-, intermediate-, and low-quality SBM did not differ, except for China, where the low-quality SBM (83.5%) had a lower (P < 0.05) digestibility than the intermediate- (89.6%) or high- (89.0%) quality meals. Soybean meal produced in Argentina (average, 87%) and Brazil (average, 82%) had lower (P < 0.05) true TAA digestibilities than did the standard SBM (91%), indicating that the processing plants in those countries may produce a less digestible SBM than that available on the open market in the United States. PMID- 15542467 TI - Effect of plant extracts and formic acid on the intestinal equilibrium of early weaned pigs. AB - We evaluated the effects of a plant extracts mixture (XT) standardized in 5% (wt/wt) carvacrol, 3% cinnamaldehyde, and 2% capsicum oleoresin (oregano, cinnamon and Mexican pepper), alone or in combination with formic acid (FA), on the productive performance and the intestinal ecosystem of the early-weaned pig. Pigs weaned at 20 +/- 1 d of age (n = 216) were allocated in 24 pens and fed a standard medicated prestarter diet for 12 d. Twelve days after weaning, a stress management system based on social and dietary stress factors was applied to the animals, after which, each group was allocated to one of six dietary treatments, which followed a factorial arrangement, with three levels (as-fed basis) of the XT (0, 150, and 300 mg/kg) and two levels of FA (0 and 0.5%). On d 24 and 25 after the stress episode, eight pigs per treatment were killed to examine variables describing some aspects of the gastrointestinal ecology. Two days after the stress episode, an Escherichia coli K88 diarrhea episode occurred, and five casualties were registered. Four of the five deaths occurred in pens of pigs not fed the XT. The FA resulted in better G:F (P = 0.040) in coincidence with shorter villous height (P = 0.073) and lower rectal total microbial mass (P = 0.078). Both XT and FA addition increased stomach content (P = 0.006 and 0.003, respectively) and percentage of DM (P = 0.089 and 0.010, respectively), suggesting an increased gastric retention time; consequently, pH was also increased (P = 0.005 and 0.060, respectively). The XT decreased ileum total microbial mass (P = 0.025) and increased the lactobacilli:enterobacteria ratio (P = 0.002). The VFA profile in the cecum and colon was modified by XT inclusion, increasing the proportion of acetate (P = 0.018 and 0.025, respectively) and diminishing the proportion of butyrate (P = 0.096 and 0.040, respectively) and valerate (P = 0.001 and 0.039, respectively). Both XT and FA were shown to be effective in modifying the gastrointestinal ecosystem, stomach contents, and stomach emptying rate, which are proposed as important aspects in the mechanisms of action for these additives. PMID- 15542468 TI - Effect of dietary crude protein level and degradability on ruminal fermentation and nitrogen utilization in lactating dairy cows. AB - The objectives of this experiment were to investigate the effects of two ruminally degradable protein (RDP) levels in diets containing similar ruminally undegradable protein (RUP) and metabolizable protein (MP) concentrations on ruminal fermentation, digestibility, and transfer of ruminal ammonia N into milk protein in dairy cows. Four ruminally and duodenally cannulated Holstein cows were allocated to two dietary treatments in a crossover design. The diets (adequate RDP [ARDP] and high RDP [HRDP]), had similar concentrations of RUP and MP, but differed in CP/RDP content. Ruminal ammonia was labeled with 15N and secretion of tracer in milk protein was determined for a period of 120 h. Ammonia concentration in the rumen tended to be greater (P = 0.06) with HRDP than with ARDP. Microbial N flow to the duodenum, ruminal digestibility of dietary nutrients, DMI, milk yield, fat content, and protein content and yield were not statistically different between diets. There was a tendency (P = 0.07) for increased urinary N excretion, and blood plasma and milk urea N concentrations were greater (P = 0.002 and P = 0.01, respectively) with HRDP compared with ARDP. Milk N efficiency was decreased (P = 0.01) by the HRDP diet. The cumulative secretion of ammonia 15N into milk protein, as a proportion of 15N dosed intraruminally, was greater (P = 0.003) with ARDP than with HRDP. The proportions of bacterial protein originating from ammonia N and milk protein originating from bacterial or ammonia N averaged 43, 61, and 26% and were not affected by diet. This experiment indicated that excess RDP in the diet of lactating dairy cows could not be efficiently utilized for microbial protein synthesis and was largely lost through urinary N excretion. At a similar MP supply, increased CP or RDP concentration of the diet would result in decreased efficiency of conversion of dietary N into milk protein and less efficient use of ruminal ammonia N for milk protein syntheses. PMID- 15542469 TI - Effects of natural plant extracts on ruminal protein degradation and fermentation profiles in continuous culture. AB - Eight dual-flow continuous culture fermenters were used in four consecutive periods of 10 d to study the effects of six natural plant extracts on ruminal protein degradation and fermentation profiles. Fermenters were fed a diet with a 52:48 forage:concentrate ratio (DM basis). Treatments were no extract (CTR), 15 mg/kg DM of a mixture of equal proportions of all extracts (MIX), and 7.5 mg/kg DM of extracts of garlic (GAR), cinnamon (CIN), yucca (YUC), anise (ANI), oregano (ORE), or pepper (PEP). During the adaptation period (d 1 through 8), samples for ammonia N and VFA concentrations were taken 2 h after feeding. On d 9 and 10, samples for VFA (2 h after feeding), and peptide, AA, and ammonia N concentrations (0, 2, 4, 6, and 8 h after feeding) were also taken. Differences were declared at P < 0.05. During the adaptation period, total VFA and ammonia N concentrations were not affected by treatments. The acetate proportion was higher from d 2 to 6 in CIN, GAR, ANI, and ORE, and the propionate proportion was lower from d 2 to 4 in CIN and GAR, and from d 2 to 5 in ANI and ORE, compared with CTR. However, the proportion of individual VFA (mol/100 mol) was similar in all treatments after d 6, except for valerate in d 9 and 10, which was lower in PEP (2.8 +/- 0.27) compared with CTR (3.5 +/- 0.27). The average peptide N concentration was 31% higher in MIX, and 26% higher in CIN and YUC compared with CTR (6.5 +/- 1.07 mg/100 mL). The average AA N concentration was 17 and 15% higher in GAR and ANI, respectively, compared with CTR (7.2 +/- 0.77 mg/100 mL). The average ammonia N concentration was 31% higher in ANI and 25.5% lower in GAR compared with CTR (5.5 +/- 0.51 mg/100 mL). The accumulation of AA and ammonia N in ANI suggested that peptidolysis and deamination were stimulated. The accumulation of AA N and the decrease in ammonia N in GAR suggests that deamination was inhibited. The accumulation of peptide N and the numerical decrease in AA N in CIN suggest that peptidolysis was inhibited. Results indicate that plant extracts modified ruminal fermentation, but microbes were adapted to some extracts after 6 d of fermentation. Therefore, data from short-term in vitro fermentation studies may lead to erroneous conclusions, and should be interpreted with caution. Careful selection of these additives may allow the manipulation of protein degradation in the rumen. PMID- 15542470 TI - Source and level of supplemental protein for growing lambs. AB - Two 3 x 2 factorial growth trials and a companion metabolism trial with 13, 15, or 17% dietary CP (DM basis), with or without 3% of the DM replaced with slowly degraded menhaden fish meal, were conducted to determine if level of dietary protein influences whether slowly degraded protein improves lamb growth and protein use. The growth trials included 32 and 34 pens of two weanling lambs initially weighing 23 to 26 kg and fed for 42 d. The metabolism trial included 12 additional lambs fed in metabolism cages with a 2-wk adjustment period, a 1-wk preliminary period, and a 7-d collection period. Plasma urea N (PUN) was measured in all lambs at the conclusion of the second growth trial and at the end of the metabolism trial. There was a protein level x protein source interaction (P = 0.05) for PUN of the 12 lambs in the metabolism trial but not for the 68 lambs in the second growth trial. Replacement of part of the soybean meal protein with protein from fish meal did not affect ADG or G:F at any protein level, but it lowered (P = 0.08) PUN in the second growth trial. Plasma urea N values were higher (P = 0.002) in lambs fed diets with 15 or 17% CP; however, ADG (P = 0.037 in Exp. 1 and P = 0.055 in Exp. 2), and G:F (P = 0.094 in Exp. 1 and P = 0.003 in Exp. 2) were lower for lambs fed the diets with 13% CP. There was little difference in ADG or G:F between lambs fed the diets with 15 or 17% CP, suggesting that a CP level of 15% with supplemental protein from soybean meal would be optimal for 25- to 40-kg growing Finnsheep x Dorset lambs. PMID- 15542471 TI - The effect of dietary ractopamine concentration and duration of feeding on growth performance, carcass characteristics, and meat quality of finishing pigs. AB - A total of 400 barrows from Dekalb EB and 83 terminal sires mated to 43 and 45 maternal lines were used to evaluate the effects of dietary ractopamine (RAC; Paylean, Elanco Animal Health, Greenfield, IN) concentrations (0, 5, 10, or 20 ppm; as-fed basis) and feeding durations (6 to 34 d) on growth, efficiency, carcass, and meat quality characteristics of finishing pigs. Barrows were weighed and sorted into five weight blocks, each block consisting of 16 pens (five pigs per pen). Weight blocks were allocated to feeding duration treatments and assigned consecutively by weight from lightest to heaviest to represent 34, 27, 20, 13, and 6 d on test, respectively. The lightest and heaviest blocks averaged 79.8 and 103.8 kg, respectively, at the start of the test. Within a weight block, pens (four per treatment) were randomly assigned to one of four dietary concentrations of RAC in a basal diet containing 18.5% CP and 1.13% lysine. The experiment-wide target slaughter weight was 109 kg, and pigs and feeders were weighed weekly. Weight blocks (80 barrows per block) were slaughtered at a commercial packing plant after 6, 13, 20, 27, or 34 d on test. Overall, RAC supplementation improved (P < 0.05) ADG; however, ADG was not different (P > 0.08) from controls for pigs fed 5, 10, and 20 ppm RAC for 27, 34, and 6 d, respectively. During each feeding period, RAC-fed pigs had improved (P < 0.05) G:F, and, after 20, 27, and 34 d on test, pigs fed 20 ppm RAC had greater (P < 0.05) G:F compared with those fed 0 or 5 ppm RAC. Hot carcass weight was increased (P < 0.05) by RAC feeding after 13 and 27 d of feeding, and by feeding 10 and 20 ppm RAC after 20 d of feeding. After 34 d, pigs fed 20 ppm RAC had heavier (P < 0.05) hot carcass weight than pigs fed 10 ppm RAC. Fat-free lean estimates and the 10th-rib LM area were increased (P < 0.05) by feeding 10 and 20 ppm RAC after 27 d, and by feeding 20 ppm RAC after 34 d compared with controls. Japanese and American color scores, as well as L*, a*, and b* values of the LM, were not affected (P > 0.11) by 5 and 10 ppm RAC compared with controls during each feeding period. Visual marbling score for the LM was decreased (P < 0.05) when RAC was fed at 10 and 20 ppm compared with 0 ppm RAC when fed for 34 d. Dietary RAC improved growth performance at all feeding durations, whereas carcass composition was improved at longer feeding durations. In addition, 5 and 10 ppm RAC did not affect objective and subjective measures of pork quality. PMID- 15542472 TI - Oxidative environments decrease tenderization of beef steaks through inactivation of mu-calpain. AB - This study was designed to test the hypothesis that oxidative conditions in postmortem (PM) tissue decrease calpain activity and proteolysis, subsequently minimizing the extent of tenderization. To achieve different levels of oxidation, the diets of beef cattle were supplemented with vitamin E for the last 126 d on feed, and beef steaks were irradiated early PM. Ten steers were fed a finishing diet with the inclusion of vitamin E at 1,000 IU per steer daily (VITE). Another 10 beef steers were fed the same finishing diet without added vitamin E (CON). At 22 to 24 h PM, strip loins from each carcass were cut into 2.54-cm-thick steaks and individually vacuum packaged. Within 26 h PM, steaks were irradiated at 0 or 6.4 kGy and then aged at 4 degrees C for 0, 1, 3, 7, and 14 d postirradiation. Steaks from each time point were used to determine Warner-Bratzler shear force (WBSF) and calpain activity, and for western blotting of sarcoplasmic proteins and myofibrillar proteins. Calpastatin activity was determined at 0, 3, and 14 d postirradiation. At 1, 3, 7, and 14 d postirradiation, WBSF values of irradiated steaks were higher (P < 0.03) than for nonirradiated steaks. Western blots of troponin-T and desmin showed decreased proteolysis in irradiated samples compared with nonirradiated samples. At 2 d PM, troponin-T degradation products were more evident (P < 0.03) in nonirradiated steaks supplemented with VITE than nonirradiated steaks from the CON diet. Similarly, VITE treatment resulted in steaks with lower (P < 0.05) calpastatin activity at 1 d PM than in steaks from steers fed the CON diet. Irradiation diminished the rate of calpastatin inactivation. Irradiated samples, regardless of diet, had no detectable levels of intact titin or nebulin. Irradiation decreased mu-calpain activity and autolysis, whereas mu-calpain activity was not affected by diet or irradiation. Inactivation of mu-calpain by oxidation during early times PM decreased the amount of myofibrillar proteolysis, thereby decreasing the extent of tenderization of beef steaks. PMID- 15542473 TI - Effect of supplemental manganese on performance and carcass characteristics of growing-finishing swine. AB - Three hundred sixteen crossbred pigs were used in two experiments to determine the effect of supplemental manganese source and dietary inclusion level during the growing-finishing period on performance and pork carcass characteristics. All pigs were blocked by weight, and treatments were assigned randomly to pens within blocks. In Exp. 1, a total of 20 pens (five pigs/pen) was randomly assigned to one of five dietary treatments consisting of control grower and finisher diets, or control diets supplemented with either 350 or 700 ppm (as-fed basis) Mn either from MnSO4 or a Mn AA complex (MnAA). In Exp. 2, a total of 36 pens (six pigs per pen) was assigned randomly to one of six dietary treatments formulated with 0, 20, 40, 80, 160, or 320 ppm (as-fed basis) Mn from MnAA. Pigs were slaughtered when the lightest block averaged 120.0 kg (Exp. 1) or at a mean BW of 106.8 kg (Exp. 2). Neither ADG nor ADFI was affected (P > 0.21) by Mn source or high inclusion level (Exp. 1); however, across the entire feeding trial, pigs consuming 320 ppm Mn from MnAA were more (P < 0.04) efficient than pigs fed diets formulated with 20 to 160 ppm Mn from MnAA (Exp. 2). Color scores did not differ (P > 0.79) at the low inclusion (20 to 320 ppm Mn) levels used in Exp. 2; however, in Exp. 1, the LM from pigs fed Mn tended to receive higher (P = 0.10) American color scores than that of pigs fed the control diet, and Japanese color scores were higher for the LM from pigs fed diets containing 350 ppm Mn from MnAA than 350 Mn from ppm MnSO4 or 700 ppm Mn from MnAA (source x inclusion level; P = 0.04; Exp. 2). Chops of pigs fed 350 ppm Mn from MnAA were darker than the LM of pigs fed 350 ppm Mn from MnSO4, and 700 ppm Mn from MnAA diets (source x inclusion level; P = 0.03; Exp. 1), but L* values were not (P = 0.76) affected by lower MnAA inclusion levels (Exp. 2). Even though the LM tended to became redder as dietary MnAA inclusion level increased from 20 to 320 ppm Mn (linear effect; P < 0.10), a* values were not (P = 0.71) altered by including 350 or 700 ppm Mn (Exp. 1). Chops of pigs fed MnAA had lower cooking losses (P = 0.01) and shear force values (P = 0.07) after 2 d of aging than did chops from pigs fed diets formulated with MnSO4. Results from these experiments indicate that feeding 320 to 350 ppm Mn from MnAA during the growing-finishing period may enhance pork quality without adversely affecting pig performance or carcass composition. PMID- 15542474 TI - Effects of dietary lysine and energy density on performance and carcass characteristics of finishing pigs fed ractopamine. AB - Two hundred sixteen crossbred barrows and gilts (84.3 kg BW) were used to test the effects of dietary energy density and lysine:energy ratio (Lys:ME) on the performance, carcass characteristics, and pork quality of finishing pigs fed 10 ppm ractopamine. Pigs were blocked by BW and gender, allotted to 36 pens (six pigs per pen), and pens were assigned randomly within blocks to dietary treatments (as-fed basis) arranged in a 2 x 3 factorial design, with two levels of energy (3.30 or 3.48 Mcal/kg) and three Lys:ME (1.7, 2.4, or 3.1 g lysine/Mcal) levels. Pigs were fed experimental diets for 28 d, and weights and feed disappearance were recorded weekly to calculate ADG, ADFI, and G:F. Upon completion of the feeding trial, pigs were slaughtered and carcass data were collected before fabrication. During carcass fabrication, hams were analyzed for lean composition using a ham electrical conductivity (TOBEC) unit, and loins were collected, vacuum-packaged, and boxed for pork quality data collection. Energy density had no (P > 0.22) effect on ADG or ADFI across the entire 28-d feeding trial; however, pigs fed 3.48 Mcal of ME were more (P < 0.02) efficient than pigs fed 3.30 Mcal of ME. In addition, ADG and G:F increased linearly (P < 0.01) as Lys:ME increased from 1.7 to 3.1 g/Mcal. Carcasses of pigs fed 3.48 Mcal of ME were fatter at the last lumbar vertebrae (P < 0.08) and 10th rib (P < 0.04), resulting in a lower (P < 0.03) predicted fat-free lean yield (FFLY). Conversely, 10th-rib fat thickness decreased linearly (P = 0.02), and LM depth (P < 0.01) and area (P < 0.01) increased linearly, with increasing Lys:ME. Moreover, FFLY (P < 0.01) and actual ham lean yield (P < 0.01) increased as Lys:ME increased in the diet. Dietary energy density had no (P > 0.19) effect on pork quality, and Lys:ME did not (P > 0.20) affect muscle pH, drip loss, color, and firmness scores. Marbling scores, as well as LM lipid content, decreased linearly (P < 0.01) as Lys:ME increased from 1.7 to 3.1 g/Mcal. There was a linear (P < 0.01) increase in shear force of cooked LM chops as Lys:ME increased in the finishing diet. Results indicate that 3.30 Mcal of ME/kg (as-fed basis) is sufficient for optimal performance and carcass leanness in pigs fed ractopamine. The Lys:ME for optimal performance and carcass composition seems higher than that currently used in the swine industry; however, feeding very high Lys:ME (> 3.0 g/Mcal, as-fed basis) to ractopamine-fed pigs may result in decreased marbling and cooked pork tenderness. PMID- 15542475 TI - Consumer visual preference and value for beef steaks differing in marbling level and color. AB - To determine visual preference and value for fresh beef steaks differing in marbling level and color, consumers in Chicago and San Francisco (n = 124 per city) evaluated two pairs of steaks in a retail case. Steaks differing in marbling level (Modest/Moderate vs. Slight) and color (bright, cherry-red vs. dark red) were purchased at retail stores in each city. Consumers selected their preferred steak in each pair, described their selection criteria, and provided the price they were willing to pay for each of the four steaks. There was a difference in visual preference in each city, with most consumers preferring (P < 0.01) low (Slight) over high (Moderate/Modest) marbling; however, more (P < 0.01) consumers in Chicago (86.7%) preferred low marbling than in San Francisco (67.0%). Selection criteria were categorized into five groups: marbling, fat, color, appearance, and palatability. Marbling was mentioned by 65.4% of consumers who preferred high marbling, whereas 64.9% of consumers who preferred low marbling mentioned fat as a selection criterion. Bright, cherry-red color was preferred by a higher (P < 0.01) percentage of consumers in both Chicago and San Francisco (67.6 and 76.5%, respectively). Color was mentioned both by consumers who preferred bright, cherry-red color (64.8%) and those who preferred dark red color (63.9%). All preference groups were willing to pay more for their preferred steak (P < 0.01), but consumers who preferred low marbling were willing to pay more (P < 0.01) for their preferred steak than consumers who preferred high marbling. Consumers who preferred bright, cherry-red color were willing to pay more (P < 0.01) for their preferred steak than consumers who preferred dark red color. Consumers who preferred low marbling seemed to desire lean products, and consumers who preferred high marbling seemed to desire products with high eating quality. In this study, consumers were willing to pay more to purchase their preferred product; however, most consumers preferred low marbling and bright, cherry-red color. PMID- 15542476 TI - Consumer sensory acceptance and value for beef steaks of similar tenderness, but differing in marbling level. AB - To determine consumer sensory acceptance and value of beef steaks differing in marbling level (high = upper USDA Choice and low = USDA Select), but similar in Warner-Bratzler shear value, consumers in Chicago and San Francisco (n = 124 per city) evaluated two matched pairs of high- and low-marbled strip steaks, and had the opportunity to participate in a silent, sealed-bid auction to purchase steaks from the same strip loins as the samples. Consumers who purchased steaks also evaluated the steaks when prepared in their homes. Based on overall acceptability ratings, consumers were categorized into three groups: 1) those who consistently found high marbling more acceptable, 2) those who consistently found low marbling more acceptable, and 3) those who were indifferent. Consumers who evaluated at least one high-marbled and one low-marbled sample in their home were included in an evaluation environment analysis (n = 50). High-marbled steaks were rated higher (P < 0.01) in juiciness, flavor, and overall acceptability than low marbled steaks. In Chicago, consumers tended to bid more (P < 0.10) for high marbled steaks, whereas consumers in San Francisco did not. Consumers who found high-marbled steaks more acceptable and those who found low-marbled steaks more acceptable were willing to pay more (P < 0.01) for the more acceptable product. Consumers who evaluated high- and low-marbled samples in both the laboratory and home environments rated high- and low-marbled samples similar (P > 0.10) in flavor, juiciness, tenderness and overall acceptability when evaluating the steaks in their homes. In addition, these consumers were willing to pay similar (P > 0.10) amounts for high- and low-marbled samples in both environments. Overall, consumers found high-marbled steaks to be more acceptable than low marbled steaks in flavor and overall acceptability when tenderness differences were minimized in the laboratory environment. Consumers were willing to pay more for their preference, whether that preference was for high-marbled or low-marbled steaks. PMID- 15542477 TI - A comparison of consumer sensory acceptance and value of domestic beef steaks and steaks from a branded, Argentine beef program. AB - To determine consumer sensory acceptance and value of branded, Argentine (grass finished, aged 30+ d) and domestic (U.S. grain-finished beef, aged 9 d) strip loins were paired based on similar Warner-Bratzler shear force values (P = 0.34) and similar marbling levels (P = 0.82). Consumers in Chicago, IL, and San Francisco, CA (n = 124 per city), evaluated one pair of Argentine and domestic steaks, and had the opportunity to participate in a silent, sealed-bid auction to purchase steaks matching the taste panel samples. Consumers were categorized into three groups based on overall acceptability ratings: 1) those who found Argentine steaks more acceptable, 2) those who found domestic steaks more acceptable, and 3) those who were indifferent. Consumers rated domestic steaks higher (P < 0.05) in juiciness, tenderness, flavor, and overall acceptability. Consumers in both Chicago and San Francisco were willing to pay more (P < 0.05) for domestic steaks (0.86 dollars and 0.52 dollars per 0.45 kg, respectively). In both cities, consumers who found Argentine samples more acceptable were willing to pay more (P < 0.05) for Argentine steaks (0.74 dollars per 0.45 kg in Chicago and 1.82 dollars per 0.45 kg in San Francisco), and consumers who found domestic samples more acceptable were willing to pay more (P < 0.05) for domestic steaks (1.66 dollars per 0.45 kg in Chicago and 1.34 dollars per 0.45 kg in San Francisco). Consumers who were indifferent were willing to pay similar (P = 0.99) amounts for Argentine and domestic steaks. Although some consumers found Argentine beef more acceptable than domestic beef (19.7 and 16.5% in Chicago and San Francisco, respectively) and were willing to pay more for it, most consumers found domestic beef to be more acceptable (59.0% in Chicago and 61.5% in San Francisco) and were willing to pay more to obtain a more acceptable product. PMID- 15542478 TI - Influence of ceftiofur sodium biobullet administration on tenderness and tissue damage in beef round muscle. AB - The effect of a biobullet (BB) containing freeze-dried ceftiofur sodium antibiotic on the presence of injection lesions, tissue damage, and histological properties, as well as Warner-Bratzler shear force (WBSF), of the biceps femoris was investigated. Steer calves (n = 25) were individually identified and assigned randomly to a product administration treatment date (7, 14, 21, 28, or 35 d before slaughter). At each pre-slaughter ceftiofur BB administration time, identified steers (n = 5) were humanely placed into a standard commercial restraining chute, where a BB implant was administered from a distance of 6.09 m. Following a standard finishing period (120 d), steers were transported to a commercial beef processing and humanely slaughtered. Following a 36-h postmortem chilling (1 degree C) period, carcasses were graded and fabricated according to industry-accepted procedures. Paired muscle samples were individually identified, collected, and aged for 14 d postmortem. Muscles were dissected into 1.27-cm strips, followed by observation and palpation for the presence of injection site lesions. Preslaughter administration times of 7 and 14 d resulted in the presence of injection lesions (80 and 20%, respectively). In addition to the control samples, no muscle damage was observed in cattle treated with BB implants 21, 28, or 35 d before slaughter. Warner-Bratzler shear force measurements taken near lesions of BB steaks and in areas 5.08 cm from lesions of control steaks tended to be higher (P < 0.10) than for other BB and control sample locations. Concentrations of insoluble and soluble collagen were higher (P < 0.05) at the site of the lesion center in lesion-afflicted vs. with control steaks. Histological determinations of the relative proportions of muscle, connective tissue, and fat were altered (P < 0.05) in BB lesion-afflicted steak cores; however, these differences were negated outside the core location of BB-treated and control steaks. It seems that using the ceftiofur BB implant system within 14 d of slaughter does create injection site lesions and increase WBSF; however, when the BB implant system, containing 100 mg of freeze-dried ceftiofur sodium, was used according to the recommended procedure (> or = 30 d preslaughter), tissue damage, alterations in histological and collagen properties, and increased meat toughness were not observed. PMID- 15542479 TI - Ionophore taste preferences of dairy heifers. AB - Two taste preference studies were conducted using six Holstein heifers in each experiment to determine preferences for no ionophore, lasalocid, or monensin in the diet. In Exp. 1, individually penned (approx. 5 mo old; 220 +/- 14 kg BW) heifers were fed a basal total mixed ration containing 46% corn silage, 46% grass haylage, and 8% soybean meal (DM basis). There were five treatments (mg/kg BW( 1)*d(-1)): 0 ionophore (control), 1 lasalocid (1L), 2 lasalocid (2L), 1 monensin (1M), or 2 monensin (2M). Ionophores were provided as part of the mineral mix that had been added to the control diet and through an ionophore-grain by-product mix to make the 2L and 2M treatments. All five diets were offered for 7 d, with the first 2 d for adaptation and the last 5 d for measurement of feed intake. The most preferred diet was then removed and the study continued with the four remaining diets. The most preferred diets were again eliminated sequentially, so that only two diets remained on d 13 and 14. Each feeding segment ranking of treatment preferences was determined based on the weight of feed refused at the end of each feeding segment. In Exp. 2, six 6-wk-old heifers (75 +/- 5 kg of BW) were individually fed either 0, 1L, or 1M in a study similar to Exp. 1, except that the most preferred diet was removed after 4 d, with the first day for adaptation and the last 3 d for measurement of feed intake. In Exp. 1, orthogonal contrasts indicated that heifers preferred the 1L and 2L diets over the 1M and 2M diets. Preferences between diet concentrations of ionophores (1 and 2 mg/kg of BW; Exp. 1) and the control and ionophore treatments did not differ, nor was there an interaction between ionophores and their concentration. Dairy heifers previously fed lasalocid prefer lasalocid over monensin when given a choice; however, heifers without previous exposure to an ionophore did not indicate a preference (Exp. 2). PMID- 15542480 TI - Cubicle housing systems for cattle: Comfort of dairy cows depends on cubicle adjustment. AB - Housing is important for the welfare of cows. Although recommendations have been proposed, abnormal movements and injury problems are still observed in cubicle houses. We conducted a survey on 70 French dairy farms that used cubicles. We examined the design of the cubicles, and the behavior, injuries, and cleanliness of the cows. Most of the cubicles did not comply with the recommendations, often being too narrow and/or too short. Difficulties in lying behavior and injuries were more common when the neck rail was high. No improvement was noted when cubicles of a recent design were used ("U.S." cubicles), apparently because these cubicles were most often cantilevered on a double head rail rather than fixed on freestanding posts. An experiment was conducted, making similar measurements, on 84 cows to compare two configurations for U.S. cubicles (cantilevered on a double head rail as observed in the survey with a high and rear neck rail vs. fixed on freestanding posts as recommended) and another recent cubicle type (Euroconfort, cantilevered on head rails, but with a large space between the rails and fixed as recommended), with and without a brisket board. In U.S. cubicles on rails, cows spent more time lying and less time fully standing inside the cubicles than in the other cubicles (percentage of time: lying, 53.9 vs. 51.5; fully standing, 7.3 vs. 8.5); in Euroconfort cubicles, they hit bars more often when getting up than in U.S. Cubicles (percentage of observations: 42.4 vs. 26.4. Without a brisket board, cows lay down more often in a fore position in U.S. cubicles than in Euroconfort ones. Somatic cell counts increased with time in U.S. cubicles on rails and decreased in the other cubicles. It is suggested that the position of the neck rail in U.S. cubicles cantilevered on rails did not leave enough space for the cow to stand inside the cubicle, thereby encouraging the cow to lie down. This could in turn favor udder contamination and/or inflammation. It is concluded that the positioning of the neck rail is of prime importance, that U.S. cubicles should be used with a brisket board and with correct positioning of the neck rail (even when a head rail is used), and that leaving a large space between head rails does not offer an adequate remedial solution for keeping a free head space in front of the cubicle. PMID- 15542481 TI - Dried skim milk as a replacement for soybean meal in growing-finishing diets: effects on growth performance, apparent total-tract nitrogen digestibility, urinary and fecal nitrogen excretion, and carcass traits in pigs. AB - Two trials were conducted to determine the replacement nutritive value of dried skim milk for growing-finishing pigs. In a three-phase feeding trial, 180 growing composite barrows (40.8 +/- 2.9 kg BW) were allotted to three dietary treatments. Each phase lasted 28 d. Treatment 1 comprised a basal corn-soybean meal diet supplemented with crystalline AA to contain true ileal digestible concentrations (as-fed basis) of 0.83, 0.66, and 0.52% Lys; 0.53, 0.45, and 0.40% Thr; and 0.51, 0.45, and 0.42% sulfur amino acids (SAA; Met + Cys) in Phases 1, 2, and 3, respectively. Treatments 2 and 3 were the basal diets with 5 and 10% (as-fed basis) dried skim milk added. The three diets at each phase were formulated to have the same quantities of DE, true ileal digestible Lys, Thr, Trp, SAA, Ca, and available P. Pigs were housed 10 per pen (six pens/treatment), allowed ad libitum access to feed, and slaughtered at 121.6 +/- 9.3 kg BW. No differences were detected between pigs fed the basal diet and the dried skim milk diets or between pigs fed the 5 and 10% dried skim milk diets, respectively, in 84-d ADG (P = 0.84 or P = 0.71), ADFI (P = 0.54 or P = 0.91), and G:F (P = 0.80 or P = 0.97), in hot carcass weight (P = 0.66 or P = 0.74), 45-min postmortem LM pH (P = 0.90 or P = 0.53), 10th-rib backfat thickness (P = 0.24 or P = 0.77), LM area (P = 0.13 or P = 0.63), weights of belly (P = 0.43 or P = 0.70), trimmed wholesale cuts (P = 0.18 to 0.85 or P = 0.06 to 0.53), and ham components (P = 0.25 to 0.98 or P = 0.32 to 0.63). In the N balance trial, four littermate pairs of finishing gilts (82.9 +/- 2.0 kg BW) were assigned within pair to the basal or the 10% dried skim milk (as-fed basis) finishing diet. Daily feed allowance was 2.6x maintenance DE requirement and was given in two equal meals. Total fecal collection from eight meals and a 96-h urine collection began on d 14 when gilts weighed 92.1 +/- 2.2 kg BW. No differences were found between dietary treatments in gilt's daily N intake (P = 0.33) and the daily output of urinary urea (P = 0.88), urinary N (P = 0.97), fecal N (P = 0.69), and total manure (P = 0.62), as well as apparent total tract N digestibility (P = 0.84) and N retention (P = 0.84). It is concluded that growing-finishing pigs fed diets containing 10% dried skim milk would have growth performance, carcass traits, and N digestibility and use similar to those fed typical corn-soybean meal diets. PMID- 15542482 TI - Methane emissions from beef cattle: Effects of monensin, sunflower oil, enzymes, yeast, and fumaric acid. AB - Methane emitted from the livestock sector contributes to greenhouse gas (GHG) emissions. Understanding the effects of diet on enteric methane production can help refine GHG emission inventories and identify viable GHG reduction strategies. Our study focused on measuring methane and carbon dioxide emissions, total-tract digestibility, and ruminal fermentation in growing beef cattle fed a diet supplemented with various additives or ingredients. Two experiments, each designed as a 4 x 4 Latin square with 21-d periods, were conducted using 16 Holstein steers (initial BW 311.6 +/- 12.3 kg). In Exp. 1, treatments were control (no additive), monensin (Rumensin, Elanco Animal Health, Indianapolis, IN; 33 mg/kg DM), sunflower oil (400 g/d, approximately 5% of DMI), and proteolytic enzyme (Protex 6-L, Genencor Int., Inc., CA; 1 mL/kg DM). In Exp. 2, treatments were control (no additive), Procreatin-7 yeast (Prince Agri Products, Inc., Quincy, IL; 4 g/d), Levucell SC yeast (Lallemand, Inc., Rexdale, Ontario, Canada; 1 g/d), and fumaric acid (Bartek Ingredients Inc., Stoney Creek, Ontario, Canada; 80 g/d). The basal diet consisted of 75% barley silage, 19% steam-rolled barley grain, and 6% supplement (DM basis). Four large chambers (two animals per chamber) were equipped with lasers and infrared gas analyzers to measure methane and carbon dioxide, respectively, for 3 d each period. Total-tract digestibility was determined using chromic oxide. Approximately 6.5% of the GE consumed was lost in the form of methane emissions from animals fed the control diet. In Exp. 1, sunflower oil decreased methane emissions by 22% (P = 0.001) compared with the control, whereas monensin (P = 0.44) and enzyme had no effect (P = 0.82). However, oil decreased (P = 0.03) the total-tract digestibility of NDF by 20%. When CH(4) emissions were corrected for differences in energy intake, the loss of GE to methane was decreased by 21% (P = 0.002) using oil and by 9% (P = 0.09) using monensin. In Exp. 2, Procreatin-7 yeast (P = 0.72), Levucell SC yeast (P = 0.28), and fumaric acid (P = 0.21) had no effect on methane emissions, although emissions as a percentage of GE intake were 3% (non-significant, P = 0.39) less for steers fed Procreatin-7 yeast compared with the control. This study demonstrates that sunflower oil, ionophores, and possibly some yeast products can be used to decrease the GE lost as methane from cattle, but fiber digestibility is impaired with oil supplementation. PMID- 15542483 TI - Effect of feed delivery fluctuations and feeding time on ruminal acidosis, growth performance, and feeding behavior of feedlot cattle. AB - Research was conducted to determine whether fluctuations in the amount of feed delivered and timing of feeding affect ruminal pH and growth of feedlot cattle. In Exp. 1, the effects of constant (C) vs. fluctuating (F) daily feed delivery on ruminal pH were assessed in a crossover experiment (two 28-d periods) involving six mature, ruminally cannulated steers. The diet consisted of 86.8% barley grain, 4.9% supplement, and 8.3% barley silage (DM basis) and was offered ad libitum for 2 wk to estimate DMI by individual steers. Steers in group C were offered a constant amount of feed daily equal to their predetermined DMI, whereas steers in group F were offered 10% more or less than their predetermined DMI on a rotating 3-d schedule. Ruminal pH of each steer was measured continuously via an indwelling electrode placed in the rumen during the last 6 d of each period. Mean pH tended to be lower (0.10 units) for F than C (5.63 vs. 5.73; P = 0.15), and ruminal pH of steers in group F tended to remain below 5.8 (P = 0.03) or 5.5 (P = 0.14) for greater proportions of the day than steers in group C. Inconsistent delivery of feed lowered ruminal pH, suggesting increased risk of subclinical acidosis. In Exp. 2, a 2 x 2 factorial was used to study the effects of pattern (C vs. F) and feeding time (morning [0900] vs. evening [2100]) on the feeding behavior and performance of 234 (310 +/- 23 kg) Charolais x Hereford beef steers during backgrounding and finishing phases over 209 d. One pen per treatment was equipped with a radio frequency identification (GrowSafe Systems Ltd., Airdrie, Canada) system that monitored bunk attendance by each steer throughout the trial. Pattern of feed delivery did not affect (P = 0.16) DMI (7.36 kg/d), ADG (1.23 kg/d), G:F (0.17), or time spent at the bunk (141 min/d), nor were pattern of feed delivery x time of feeding interactions observed (P = 0.18). Late feeding increased (P < 0.05) daily DMI (7.48 vs. 7.26 kg), ADG (1.28 vs. 1.00 kg/d), and G:F (0.21 vs. 0.15). These studies indicate that the risk of subclinical acidosis was increased with fluctuating delivery of feed, but the greater risk of acidosis did not impair growth performance by feedlot cattle. Consequently, daily intake fluctuations of 10% DMI or less that do not alter overall intake by feedlot cattle are unlikely to have any negative consequences on growth performance. PMID- 15542484 TI - The relationship of average backfat thickness of feedlot steers to performance and relative efficiency of fat and protein retention. AB - Selection for growth and improved carcass merit has resulted in cattle that are variable in composition of gain during the finishing phase. This study assessed the relative performance among cattle with different levels of initial backfat thickness. It also exploited the ability to track carcass composition in the live animal with ultrasound estimates of backfat and marbling. A procedure was developed to partition and estimate relative efficiency of fat and protein gain. The trial periods were the last 43 or 50 d before slaughter and included 10 pens (average of 27 animals per pen) that ranged in average backfat thickness from 6.3 to 13.1 mm. There was no correlation (r2 = 0.0026) between average backfat thickness and G:F (g/kg of DMI). Correlations between average backfat thickness and ADG or DMI were also nearly zero (r2 = 0.0007 and 0.0042, respectively). Fat deposition from NEg was 3.98 times more efficient than protein deposition. Carcass backfat thickness was a poor predictor of carcass marbling score (r2 = 0.083), even though backfat thickness was an important predictor of the percentage of empty body fat (r2 = 0.807). The results indicate that a measure of backfat thickness on the live animal during the finishing phase is not an effective predictor of future feed efficiency. They also confirm that protein accretion is energetically expensive, and that using a single coefficient for predicting gain from NEg is valid regardless of whether gain is predominantly muscle or fat. These data document that there is little relationship between body composition and marbling score, which is contrary to models that assume a USDA quality grade target at a specified percent fat end point. PMID- 15542485 TI - Response of barrows to space allocation and ractopamine. AB - An experiment using 264 crossbred barrows was conducted to examine the interaction between space allocation and dietary ractopamine addition on pig performance and carcass characteristics using a 2 x 2 factorial arrangement of treatments. Treatments were 0.55 (19 pigs per pen) or 0.74 (14 pigs per pen) m2/pig from start (29.7 +/- 0.1 kg BW) to slaughter (108 kg BW) in a fully slatted facility and 0 or 10 ppm (as-fed basis) ractopamine for 28 d before slaughter. There were few treatment interactions. Pigs given 0.55 m2/pig had a lower ADG (P = 0.010), ADFI (P = 0.088), 10th-rib backfat depth on d 86 (P = 0.010), and carcass loin muscle depth (P = 0.011) than pigs given 0.74 m2/pig. There was no difference in feed conversion (P = 0.210) as a result of space allocation. Pigs fed diets containing 10 ppm ractopamine had decreased (P = 0.004) ADFI and improved (P = 0.001) feed conversion efficiencies for the 28-d feeding period, along with greater loin depth (P = 0.005) and carcass lean percent (P = 0.001). The improvements in 28-d carcass lean growth associated with feeding 10 ppm ractopamine resulted in an improvement in overall daily fat-free lean gain (P = 0.046). Under these experimental conditions, the response to dietary ractopamine was similar for crowded and uncrowded pigs. PMID- 15542486 TI - Testosterone and luteinizing hormone responses to naloxone help predict sexual performance in rams. AB - The first objective of this study was to determine whether LH and testosterone respond differently to a naloxone injection in relation to varying sexual performance in rams. If differences occurred, the second objective was to determine whether differences would predict variation in sexual performance. From a group of 1.5- to 3-yr-old rams, 20 sexually active and 39 sexually inactive rams were selected based on previously observed sexual behavior with estrual ewes. Each ram was exposed to three estrual ewes for 18 30-min sexual performance tests, and those found to be inactive were given two 30-min sexual partner preference tests. The final distribution was 28 sexually active, 22 inactive, and nine male-oriented rams. Rams were treated with 1.5 mg of naloxone/kg BW in December of Year 1 and again with either 0.75 or 1.5 mg of naloxone/kg BW in November of Year 2. Plasma concentrations of LH and testosterone were evaluated with mixed model analyses for repeated measures separately for each year to coincide with logistic procedures for modeling the probability that rams were sexually active. For Year 1, a sexual activity x age x time interaction for LH after naloxone was observed (P < 0.03). For testosterone, there was a sexual activity x time interaction (P < 0.03), with a similar, early increase for sexually active female-and male-oriented rams compared with a delayed, minimal increase for inactive rams. For Year 2, when all rams were over 2.5 yr of age, a sexual activity x time interaction for both LH and testosterone (P < 0.02) seemed more related to an earlier increase of both hormones for sexually active rams than the increase observed for inactive rams. In addition, sexually active rams had a greater increase in testosterone than inactive rams. No significant difference was observed between 0.75 and 1.50 mg of naloxone/kg BW. Testosterone and LH were used as explanatory variables and sexual activity was used as the response variable in logistic procedures. In Year 1, greatest prediction accuracy was 73.5% using testosterone at 60 min after naloxone injection. In Year 2, the greatest prediction accuracy was 85% using LH at 15 min multiplied by testosterone at 60 min after naloxone. Test repeatability for both years on the same rams was 76%. In conclusion, pattern and magnitude of naloxone-induced changes in endocrine function may facilitate identification of sexually active and inactive rams during the breeding season. Prediction accuracy of the naloxone based test was 69 to 85%. PMID- 15542487 TI - Productivity of cow-calf pairs grazing tall fescue pastures infected with either the wild-type endophyte or a nonergot alkaloid-producing endophyte strain, AR542. AB - The nonergot alkaloid-producing endo-phyte, AR542, has been shown to improve the persistence and yield of tall fescue pastures without causing the animal disorders commonly associated with tall fescue toxicosis. A 3-yr grazing study was conducted to compare effects of AR542-infected tall fescue pastures with wild type endophyte-infected (E+) tall fescue pastures on cow-calf performance. Replicated 7.3-ha pastures of each treatment were grazed by cow-calf pairs (16 pairs per pasture replication) each year from March to weaning in September. The cows were exposed to breeding on their respective pasture treatments from April 1 through June 15. The treatment groups were compared for reproductive performance, ADG, BCS, calf growth rate, and weaning weight. Blood samples were also collected for serum prolactin (PRL) analysis. There were no significant differences in calving rate (P = 0.98) or calving interval (P = 0.62) between pasture treatments. Cows that grazed the AR542 pastures subsequently gave birth to calves that were heavier (P < 0.05) than calves from cows that had grazed the E+ pastures. Cows grazing the AR542 pastures had higher (P < 0.05) BCS at the end of the grazing period, and had higher ADG during the grazing period. Calves raised on the AR542 pasture had higher (P < 0.05) ADG and weaning weights than calves of the same sex raised on the E+ pastures. Serum PRL concentrations were decreased (P < 0.05) in both cows and calves on the E+ pastures compared with serum PRL concentrations in cows and calves grazing the AR542 pastures. The results indicate that grazing tall fescue pastures infected with the AR542 endophyte may give significant advantages in cow-calf growth rates and BCS over grazing E+ pastures. However, there did not seem to be any benefit in reproductive performance in this trial. There was a small, but significant increase in birth weight in cows grazing AR542 pasture. PMID- 15542488 TI - An unaddressed issue of agricultural terrorism: a case study on feed security. AB - In the late winter of 2003, a number of livestock animals in the Midwest were poisoned due the accidental contamination of a popular commercial feed with a lethal additive. Although all the evidence indicates this incident had no malicious or terrorist intent, it is informative as a case study highlighting potential security implications with respect to a terrorist event directed at U.S. agriculture. PMID- 15542489 TI - Rice contains two disparate ent-copalyl diphosphate synthases with distinct metabolic functions. AB - Rice (Oryza sativa) produces ent-copalyl diphosphate for both gibberellin (GA) phytohormone and defensive phytoalexin biosynthesis, raising the question of how this initial biosynthetic step is carried out for these distinct metabolic processes. Here, a functional genomics approach has been utilized to identify two disparate ent-copalyl diphosphate synthases from rice (OsCPS1ent and OsCPS2ent). Notably, it was very recently demonstrated that only one of these (OsCPS1ent) normally operates in GA biosynthesis as mutations in this gene result in severely impaired growth. Evidence is presented here strongly indicating that the other (OsCPS2ent) is involved in related secondary metabolism producing defensive phytochemicals. In particular, under appropriate conditions, OsCPS2ent mRNA is specifically induced in leaves prior to production of the corresponding phytoalexins. Thus, transcriptional control of OsCPS2ent seems to be an important means of regulating defensive phytochemical biosynthesis. Finally, OsCPS1ent is significantly more similar to the likewise GA-specific gene An1/ZmCPS1ent in maize (Zea mays) than its class II terpene synthase paralogs involved in rice secondary metabolism. Hence, we speculate that this cross-species conservation by biosynthetic process reflects derivation of related secondary metabolism from the GA primary biosynthetic pathway prior to the early divergence between the separate lineages within the cereal/grass family (Poaceae) resulting in modern rice and maize. PMID- 15542490 TI - Organization of monoterpene biosynthesis in Mentha. Immunocytochemical localizations of geranyl diphosphate synthase, limonene-6-hydroxylase, isopiperitenol dehydrogenase, and pulegone reductase. AB - We present immunocytochemical localizations of four enzymes involved in p menthane monoterpene biosynthesis in mint: the large and small subunits of peppermint (Mentha x piperita) geranyl diphosphate synthase, spearmint (Mentha spicata) (-)-(4S)-limonene-6-hydroxylase, peppermint (-)-trans-isopiperitenol dehydrogenase, and peppermint (+)-pulegone reductase. All were localized to the secretory cells of peltate glandular trichomes with abundant labeling corresponding to the secretory phase of gland development. Immunogold labeling of geranyl diphosphate synthase occurred within secretory cell leucoplasts, (-)-4S limonene-6-hydroxylase labeling was associated with gland cell endoplasmic reticulum, (-)-trans-isopiperitenol dehydrogenase labeling was restricted to secretory cell mitochondria, while (+)-pulegone reductase labeling occurred only in secretory cell cytoplasm. We discuss this pathway compartmentalization in relation to possible mechanisms for the intracellular movement of monoterpene metabolites, and for monoterpene secretion into the extracellular essential oil storage cavity. PMID- 15542491 TI - Promoter analysis of the barley Pht1;1 phosphate transporter gene identifies regions controlling root expression and responsiveness to phosphate deprivation. AB - Previous studies have shown that the promoter from the barley (Hordeum vulgare) phosphate transporter gene, HvPht1;1, activates high levels of expression in rice (Oryza sativa) roots and that the expression level was induced by up to 4-fold in response to phosphorus (P) deprivation. To identify promoter regions controlling gene regulation specificities, successive promoter truncations were made and attached to reporter genes. Promoters of between 856 and 1,400 nucleotides activated gene expression in a number of cell types but with maximal expression in trichoblast (root hair) cells. For shorter promoters the trichoblast specificity was lost, but in other tissues the distribution pattern was unchanged. The low P induction response was unaffected by promoter length. Domain exchange experiments subsequently identified that the region between -856 and 547 nucleotides (relative to the translational start) is required for epidermal cell expression. A second region located between 0 and -195 nucleotides controls root-tip expression. The HvPht1;1 promoter contains one PHO-like motif and three motifs similar to the dicot P1BS element. Analysis of promoters from which the PHO-like element was eliminated (by truncation) showed no change in the gene induction response to P deficiency. In contrast, mutation of the P1BS elements eliminated any induction of gene expression in response to low P. An internal HvPht1;1 promoter fragment, incorporating a single P1BS element, had an increased response to P deprivation in comparison with the unmodified promoter (containing three elements). Together these findings further our understanding of the regulation of the HvPht1;1 gene and provide direct evidence for a functional role of the P1BS element in the expression of P-regulated genes. PMID- 15542492 TI - Ca2+-permeable channels in the plasma membrane of Arabidopsis pollen are regulated by actin microfilaments. AB - Cytosolic free Ca2+ and actin microfilaments play crucial roles in regulation of pollen germination and tube growth. The focus of this study is to test the hypothesis that Ca2+ channels, as well as channel-mediated Ca2+ influxes across the plasma membrane (PM) of pollen and pollen tubes, are regulated by actin microfilaments and that cytoplasmic Ca2+ in pollen and pollen tubes is consequently regulated. In vitro Arabidopsis (Arabidopsis thaliana) pollen germination and tube growth were significantly inhibited by Ca2+ channel blockers La3+ or Gd3+ and F-actin depolymerization regents. The inhibitory effect of cytochalasin D (CD) or cytochalasin B (CB) on pollen germination and tube growth was enhanced by increasing external Ca2+. Ca2+ fluorescence imaging showed that addition of actin depolymerization reagents significantly increased cytoplasmic Ca2+ levels in pollen protoplasts and pollen tubes, and that cytoplasmic Ca2+ increase induced by CD or CB was abolished by addition of Ca2+ channel blockers. By using patch-clamp techniques, we identified the hyperpolarization-activated inward Ca2+ currents across the PM of Arabidopsis pollen protoplasts. The activity of Ca2+-permeable channels was stimulated by CB or CD, but not by phalloidin. However, preincubation of the pollen protoplasts with phalloidin abolished the effects of CD or CB on the channel activity. The presented results demonstrate that the Ca2+-permeable channels exist in Arabidopsis pollen and pollen tube PMs, and that dynamic actin microfilaments regulate Ca2+ channel activity and may consequently regulate cytoplasmic Ca2+. PMID- 15542493 TI - ER-derived compartments are formed by highly regulated processes and have special functions in plants. PMID- 15542494 TI - Targeting of proteins to endoplasmic reticulum-derived compartments in plants. The importance of RNA localization. PMID- 15542495 TI - Protein quality control mechanisms and protein storage in the endoplasmic reticulum. A conflict of interests? PMID- 15542496 TI - Endoplasmic reticulum, oleosins, and oils in seeds and tapetum cells. PMID- 15542497 TI - Diversity and formation of endoplasmic reticulum-derived compartments in plants. Are these compartments specific to plant cells? PMID- 15542498 TI - Endoplasmic reticulum to vacuole trafficking of endoplasmic reticulum bodies provides an alternate pathway for protein transfer to the vacuole. PMID- 15542499 TI - Transcription factor networks. Pathways to the knowledge of root development. PMID- 15542500 TI - Sequence and comparative analysis of the maize NB mitochondrial genome. AB - The NB mitochondrial genome found in most fertile varieties of commercial maize (Zea mays subsp. mays) was sequenced. The 569,630-bp genome maps as a circle containing 58 identified genes encoding 33 known proteins, 3 ribosomal RNAs, and 21 tRNAs that recognize 14 amino acids. Among the 22 group II introns identified, 7 are trans-spliced. There are 121 open reading frames (ORFs) of at least 300 bp, only 3 of which exist in the mitochondrial genome of rice (Oryza sativa). In total, the identified mitochondrial genes, pseudogenes, ORFs, and cis-spliced introns extend over 127,555 bp (22.39%) of the genome. Integrated plastid DNA accounts for an additional 25,281 bp (4.44%) of the mitochondrial DNA, and phylogenetic analyses raise the possibility that copy correction with DNA from the plastid is an ongoing process. Although the genome contains six pairs of large repeats that cover 17.35% of the genome, small repeats (20-500 bp) account for only 5.59%, and transposable element sequences are extremely rare. MultiPip alignments show that maize mitochondrial DNA has little sequence similarity with other plant mitochondrial genomes, including that of rice, outside of the known functional genes. After eliminating genes, introns, ORFs, and plastid-derived DNA, nearly three-fourths of the maize NB mitochondrial genome is still of unknown origin and function. PMID- 15542502 TI - Increased gut permeability in Crohn's disease: is TNF the link? PMID- 15542503 TI - Recurrent bile duct stones after endoscopic sphincterotomy. PMID- 15542504 TI - Molecular diagnosis of pancreatobiliary malignancies in brush cytologies of biliary strictures. PMID- 15542505 TI - Lifestyle related risk factors in the aetiology of gastro-oesophageal reflux. AB - BACKGROUND/AIM: The aetiology of gastro-oesophageal reflux is largely unknown. The authors' aim was to examine the relation between lifestyle habits and gastro oesophageal reflux symptoms. SUBJECTS: Participants of two consecutive public health surveys in Nord-Trondelag, Norway. METHODS: In a case control study within the two public health surveys, 3153 individuals who in the second survey reported severe heartburn or regurgitation during the last 12 months were defined as cases, while 40 210 people without reflux symptoms constituted the control group. The risk of reflux symptoms was estimated and multivariately calculated as odds ratios in relation to exposure to tobacco smoking, alcohol, coffee, tea, table salt, cereal fibres, and physical exercise. RESULTS: There was a significant dose response association between tobacco smoking and reflux symptoms. Among people who had smoked daily for more than 20 years the odds ratio was 1.7 (95% confidence interval 1.5 to 1.9) compared with non-smokers. A similar positive association was found for table salt intake. The odds ratio for reflux was 1.7 (95% CI 1.4 to 2.0) among those who always used extra table salt compared with those who never did so. We found moderately strong negative associations between the risk of reflux and exposure to coffee, bread high in dietary fibre content, and frequent physical exercise. Intake of alcohol or tea did not affect the risk of reflux. CONCLUSIONS: Tobacco smoking and table salt intake seem to be risk factors for gastro-oesophageal reflux symptoms. Dietary fibres and physical exercise may protect against reflux. Alcohol, coffee, and tea do not seem to be risk factors for reflux. PMID- 15542507 TI - Brown spots in the bowel. PMID- 15542508 TI - Endoluminal gastroplication in children with significant gastro-oesophageal reflux disease. AB - AIMS: To describe paediatric experience, and to assess complications and therapeutic effectiveness of the use of endoluminal gastroplication in children with gastro-oesophageal reflux disease (GORD) refractory to, or dependent on, proton pump inhibitors. METHODS: Seventeen (five male) consecutive children/adolescents (median (range) age 12.4 (6.1-15.9) years, median (range) weight 46.0 (16.5-87.5) kg) with GORD either dependent for more than 12 months on proton pump inhibitors or non-responsive to medical treatment underwent endoscopic gastroplication using a flexible endoscopic sewing device (EndoCinch). Three plications were placed in gastric tissue below the lower oesophageal sphincter. Drug dose requirement, pH measurements, daily symptom severity and frequency, and validated reflux (QOLRAD) and general gastrointestinal (GSRS) quality of life scores were compared before and after endoscopic gastroplication. RESULTS: All patients showed post-treatment improvement in symptom severity, frequency, and quality of life scores (p<0.0001). Three patients with recurrent symptomatic GORD had a repeat procedure within six weeks and did well subsequently. At up to 33 months of follow up (median 23), 14/17 patients remained off all antireflux medications, and 14/17 had maintained their symptomatic improvement. All pH parameters improved and had returned to normal values in 14/16 patients post-treatment and in 6/9 after one year of follow-up: in particular the reflux index had decreased from a median of 16.6% (0.9-67%) to 2.5% (0.7-15.7%) (p<0.0001) six weeks and 4.3% (2.2-20.6) (p<0.02) 12 months post procedure. The only complication observed was gastric bleeding in one patient due to previously undiagnosed coagulopathy, which spontaneously resolved. CONCLUSIONS: Endoluminal gastroplication is an effective and safe procedure in children/adolescents with significant GORD refractory to, or dependent on, medical anti-GORD therapy. PMID- 15542506 TI - What is the best management strategy for high grade dysplasia in Barrett's oesophagus? A cost effectiveness analysis. AB - BACKGROUND: Multiple treatment strategies for subjects with high grade dysplasia (HGD) in Barrett's oesophagus (BO) have been suggested. However, it is unclear which of these strategies provides the greatest life expectancy, and the costs associated with the management strategies are unknown. AIM: To compare the efficacy and cost effectiveness of competing management strategies for BO with HGD. METHODS: We created a decision analysis model in Data 4.0 to assess possible treatment strategies for BO with HGD. The strategies included: (1) no preventative strategy, (2) elective surgical oesophagectomy, (3) endoscopic ablation, and (4) surveillance endoscopy. The base case was a healthy 50 year old White male with an initial diagnosis of BO with HGD. The model allowed for complications of surgery, including death. Ablative therapy could cause stricture or perforation. Pathological misinterpretation was allowed, and modelled after reported rates. Estimates were derived from the literature for the rate of progression of HGD to cancer and for complication rates for the various treatment modalities. The endoscopic ablation arm was modelled as photodynamic therapy. Sensitivity analyses were performed over a wide range of cancer incidences, complication rates, and procedure costs. RESULTS: Endoscopic ablation was the most effective strategy, yielding 15.5 discounted quality adjusted life years (dQALY), compared with 15.0 for endoscopic surveillance and 14.9 for oesophagectomy. No preventative strategy was the most inexpensive option, yielding an average cost per quality adjusted life year of US dollars 54 (44) per dQALY, but resulted in high rates of cancer. Endoscopic surveillance dominated oesophagectomy, being both less costly and more effective. The condition of extended dominance occurred when comparing endoscopic ablation to endoscopic surveillance because, although the total costs of ablation were greater than those of surveillance, it was less expensive to buy an additional life year using endoscopic ablation than endoscopic surveillance. The incremental cost effectiveness ratio when moving from no therapy to ablative therapy was a reasonable US dollars 25 621/dQALY (21 009/dQALY). Sensitivity analysis demonstrated that when yearly rates of progression to cancer from HGD exceeded 30%, oesophagectomy became the most cost effective option. CONCLUSIONS: A strategy of endoscopic ablation provided the longest quality adjusted life expectancy for BO with HGD. Although endoscopic surveillance was less expensive than endoscopic ablation, it was associated with shorter survival. Optimal utilisation of healthcare resources may be achieved with endoscopic ablative therapy for BO with HGD. PMID- 15542510 TI - Helicobacter pylori test and eradicate versus prompt endoscopy for management of dyspeptic patients: 6.7 year follow up of a randomised trial. AB - BACKGROUND: Dyspepsia is a chronic disease with significant impact on the use of health care resources. A management strategy based on Helicobacter pylori testing has been recommended but the long term effect is unknown. AIM: To investigate the long term effect of a test and treat strategy compared with prompt endoscopy for management of dyspeptic patients in primary care. PATIENTS: A total of 500 patients presenting in primary care with dyspepsia were randomised to management by H pylori testing plus eradication therapy (n = 250) or by endoscopy (n = 250). Results of 12 month follow up have previously been presented. METHODS: Symptoms, quality of life, and patient satisfaction were recorded during a three month period, a median 6.7 years after randomisation (range 6.1-7.3 years). Number of endoscopies, antisecretory medication, H pylori treatments, and hospital visits were recorded from health care databases for the entire follow up period. RESULTS: Median age was 45 years; 28% were H pylori infected. Use of resources was registered in all 500 patients (3084 person years) of whom 312 completed diaries. We found no difference in symptoms between the two groups. Median proportion of days without symptoms was 0.52 (interquartile range 0.10-0.88) in the test and eradicate group versus 0.64 (0.14-0.90) in the prompt endoscopy group (p = 0.27) (mean difference 0.05 (95% confidence interval (CI) -0.03 to 0.14)). Compared with the prompt endoscopy group, the test and eradicate group underwent fewer endoscopies (mean difference 0.62 endoscopies/person (95% CI 0.38 0.86)) and used less antisecretory medication (mean difference 102 defined daily doses/person (95% CI -1 to 205)). CONCLUSION: On a long term basis, a H pylori test and eradicate strategy is as efficient as prompt endoscopy for management of dyspeptic patients in primary care and reduces the use of endoscopy and antisecretory medication. PMID- 15542511 TI - Acetylated sialic acid residues and blood group antigens localise within the epithelium in microvillous atrophy indicating internal accumulation of the glycocalyx. AB - BACKGROUND: Microvillous atrophy, a disorder of intractable diarrhoea in infancy, is characterised by the intestinal epithelial cell abnormalities of abnormal accumulation of periodic acid-Schiff (PAS) positive secretory granules within the apical cytoplasm and the presence of microvillous inclusions. The identity of the PAS positive material is not known, and the aim of this paper was to further investigate its composition. METHODS: Formaldehyde fixed sections were stained with alcian blue/PAS to identify the acidic or neutral nature of the material, phenylhydrazine blocking was employed to stain specifically for sialic acid, and saponification determined the presence of sialic acid acetylation. The specificity of sialic acid staining was tested by digestion with mild sulphuric acid. Expression of blood group related antigens was tested immunochemically. RESULTS: Alcian blue/PAS staining identified a closely apposed layer of acidic material on the otherwise neutral (PAS positive) brush border in controls. In microvillous atrophy, a triple layer was seen with an outer acidic layer, an unstained brush border region, and accumulation within the epithelium of a neutral glycosubstance that contained acetylated sialic acid. Blood group antigens were detected on the brush border, in mucus, and within goblet cells in controls. In microvillous atrophy they were additionally expressed within the apical cytoplasm of epithelial cells mirroring the PAS abnormality. Immuno electron microscopy localised expression to secretory granules. CONCLUSIONS: A neutral, blood group antigen positive, glycosubstance that contains acetylated sialic acid accumulates in the epithelium in microvillous atrophy. Previous studies have demonstrated that the direct and indirect constitutive pathways are intact in this disorder and it is speculated that the abnormal staining pattern reflects accumulation of glycocalyx related material. PMID- 15542509 TI - Cyclooxygenase 1 is required for pH control at the mouse gastric surface. AB - BACKGROUND: Endogenous cyclooxygenase (COX) activity is required to maintain a relatively alkaline surface pH at the gastric luminal surface. AIMS: The purpose of this study was to determine which COX isoform, COX-1 or COX-2, is responsible for regulating the protective surface pH gradient and to test if COX inhibitors also had non-COX mediated effects in vivo. METHODS: Immunofluorescence and western blot analysis showed constitutive expression of both COX isoforms in the normal mouse stomach. We used in vivo confocal microscopy to measure pH near the mucosal surface of anaesthetised COX-1 (-/-), COX-2 (-/-), or wild-type mice of the same genetic background. RESULTS: When the gastric mucosal surface was exposed and superfused (0.2 ml/min) with a weakly buffered saline solution (pH 3) containing the pH indicator Cl-NERF, the pH directly at the gastric surface and thickness of the pH gradient were similar in wild-type and COX-2 (-/-) mice, but COX-1 (-/-) mice had a significantly thinner pH gradient. Addition of indomethacin had minimal effects on the residual surface pH gradient in COX-1 (-/ ) mice, suggesting no role for COX-2 in surface pH regulation. Whole stomach perfusion studies demonstrated diminished net alkali secretion in COX-1 (-/-) mice, and application of SC-560 or rofecoxib to wild-type mice and mutant mice confirmed that only COX-1 inhibition reduced alkali secretion. CONCLUSION: COX-1 is the dominant isoform regulating the normal thickness of the protective surface pH gradient in mouse stomach. PMID- 15542512 TI - Unexplained digestive bleeding in a cirrhotic patient. PMID- 15542513 TI - Depletion of intestinal resident macrophages prevents ischaemia reperfusion injury in gut. AB - BACKGROUND AND AIMS: The cellular and molecular events involved in ischaemia reperfusion (IR) injury are complex and not fully understood. Previous studies have implicated polymorphonuclear neutrophils (PMN) as major inflammatory cells in IR injury. However, anti-PMN antiserum treatment offers only limited protection, indicating that other inflammatory cells are involved. We have therefore investigated the contribution of resident macrophages in IR injury using an IR gut injury model. METHODS: DA rats were divided into sham operation and IR groups. The superior mesenteric artery was clamped for 30, 45, or 60 minutes (ischaemia) followed by 60 minutes of reperfusion. IR injuries were evaluated by histological staining. Expression of early growth response factor 1 (Egr-1), myeloperoxidase (MPO), and proinflammatory cytokines was analysed by immunohistochemistry, reverse transcription-polymerase chain reaction, and western blotting analysis. The specific role of macrophages in IR gut injury was also evaluated in resident macrophage depleted rats. RESULTS: Mucosal sloughing and villi destruction were seen in 45/60 minute and 60/60 minute IR guts. PMN infiltration at the damaged mucosal area was undetectable in 45/60 minute and 60/60 minute IR guts. PMN were localised around the capillaries at the base of the crypts in 60/60 minute IR gut. Obvious PMN infiltration was only observed in damaged villi after three hours of reperfusion. Elevated nuclear Egr-1 immunostaining was localised in resident macrophages at the damaged villi before histological appearance of mucosal damage. Furthermore, resident macrophages at the damaged site expressed MPO. Protein levels of the proinflammatory cytokines RANTES and MCP-1 were increased in IR gut. Depletion of resident macrophages by dichloromethylene bisphosphonate significantly reduced mucosal damage in rat guts after IR. CONCLUSION: Our findings indicate that resident macrophages play a role in early mucosal damage in IR gut injury. Therefore, macrophages should be treated as a prime target for therapeutic intervention for IR damage. PMID- 15542514 TI - Citrulline increases arginine pools and restores nitrogen balance after massive intestinal resection. AB - OBJECTIVE: Arginine supplementation seems logical in situations where this amino acid becomes essential, for example after massive intestinal resection. Arginine is taken up and metabolised by the liver to a large extent and its supplementation is potentially unsafe. Citrulline is not captured by the liver and passes freely to the kidneys where it is metabolised to arginine, and so is a good candidate to generate arginine and thereby improve nutritional status. METHODS: Twenty four rats were assigned to four groups: citrulline, arginine, control, and sham. The sham group underwent transection and the three other groups resection of 80% of the small intestine. All rats were fed by enteral nutrition and its composition was as follows: supplementation with citrulline in the citrulline group, supplementation with arginine in the arginine group, and standard polymeric enteral nutrition in the control and sham groups. All groups received isonitrogenous nutrition and citrulline and arginine intakes were equimolar in their respective groups. After 10 days, the rats were sacrificed. RESULTS: Arginine concentration was higher (p<0.05) in plasma and muscle in the citrulline group than in the three other groups. Plasma levels of arginine were 110 (12), 79 (7), 167 (22), and 228 (13) mumol/l in the sham, control, arginine, and citrulline groups respectively. Arginine concentrations in the gastrocnemius were: 0.15 (0.02), 0.16 (0.02), 0.40 (0.05), and 0.94 (0.20) mumol/g, respectively. Citrulline preserved nitrogen balance in resected rats but not in arginine supplemented rats (mean J10: 2.27 (0.29), 1.67 (0.15), 1.98 (0.29), and 2.43 (0.41) g/24 hours in the sham, control, arginine, and citrulline groups, respectively). CONCLUSION: Supplementing the diet with citrulline is a very efficient means of increasing arginine levels and improving nitrogen balance after massive intestinal resection. The results of this study form a strong rationale for citrulline supplementation in these patients. PMID- 15542515 TI - The role of tension receptors in colonic mechanosensitivity in humans. AB - BACKGROUND: Perception of colonic distension, which is enhanced in a subset of patients with irritable bowel syndrome, requires activation of mechanoreceptors. In animal studies, distension activates both in series ("tension") and in parallel ("elongation") mechanoreceptors. During active contractions against a fixed volume balloon, tension receptors are activated without elongation of receptor activation. AIM: To evaluate the role of tension receptors in the perception of mechanical stimuli from the colon in healthy subjects. METHODS: A 700 ml balloon connected to a barostat-manometer assembly was placed in the descending colon of 10 healthy subjects. After volume controlled distension (50 ml/2 minutes) to assess the first perception threshold, fixed volume subthreshold distension (122 (16) ml) was maintained for a 30 minute period before and after administration of neostigmine 0.5 mg intravenously. Mean intraballoon pressure, number, amplitude, and duration of contractions, and frequency of sensations were analysed. The period after neostigmine was divided into 10 second intervals and evaluated for the occurrence of contractions and onset of sensations. Fisher's exact test was applied to calculate the sensation-contraction association probability (SAP) as (1.0-p)x100%. RESULTS: Neostigmine increased intraballoon pressure (p<0.01), number of contractions (p<0.01), and number of sensations (p<0.01) per minute in all subjects. In seven of 10 subjects a significant association (SAP >95%) was found between sensations and contractions. In the remaining subjects, contractions were not associated with sensations and had lower amplitude (p<0.05) and duration (p<0.01) compared with contractions in the other seven subjects. CONCLUSION: In humans, tension receptors are involved in mediating colonic mechanosensitivity. PMID- 15542516 TI - Complication of Waldenstroem's macroglobulinaemia following ERCP. PMID- 15542517 TI - Acute tryptophan depletion affects brain-gut responses in irritable bowel syndrome patients and controls. AB - BACKGROUND: Serotonin, a key denominator of the brain-gut axis, is involved in the regulation of gastrointestinal motility, secretion, and perception as well as cognition and mood. AIM: To assess the effects of an acutely lowered serotonin synthesis, using the acute tryptophan depletion (ATD) method, on visceral perception, affective memory performance, and mood in diarrhoea predominant irritable bowel syndrome patients (d-IBS) and controls. METHODS: In a randomised, double blind, crossover design, 14 d-IBS patients and fourteen matched controls were studied under ATD and placebo conditions, respectively. Perception of urge and pain was scored during rectal distensions. Affective memory performance, mood, and biochemical parameters of serotonergic metabolism were simultaneously assessed. RESULTS: ATD significantly decreased plasma tryptophan (67.0 (2.0) v 24.9 (2.0) mumol/l) and 5-hydroxyindole acetic acid concentrations (29.9 (1.0) v 15.8 (0.6) nmol/l). ATD was associated with significantly increased urge scores specifically in the lower pressure range and overall increased pain scores. ATD significantly lowered the perceptual threshold for first perception compared with placebo (patients 10.6 (1.2) v 13.6 (0.8) mm Hg, controls 12.6 (1.3) v 15.7 (1.2) mm Hg) but not for maximal tolerable discomfort (patients 50.5 (3.6) v 51.6 (3.3) mm Hg, controls 50.9 (3.3) v 48.8 (2.9) mm Hg). ATD induced a significant shift in affective memory bias towards preferential loss of positive material but no significant changes in mood. ATD did not differentially affect the patient or control group. CONCLUSIONS: We have provided evidence that serotonergic modulation by ATD affects both visceral perception as well as cognition in d-IBS and controls. Simultaneous measurement of brain and gut function and the application of ATD contribute to the elucidation of the complex pathophysiology of IBS. PMID- 15542518 TI - Human lymphocyte stimulation with pouchitis flora is greater than with flora from a healthy pouch but is suppressed by metronidazole. AB - BACKGROUND/AIMS: The gut flora may play an important role in the pathogenesis of inflammatory bowel disease. An ileal reservoir or pouch can be created to replace the excised rectum after proctocolectomy. In patients with ulcerative colitis this is subject to inflammation and termed pouchitis. Using bacteria from patients the authors sought evidence for the presence rather than the identity of a pathogenic species in pouchitis, and for its absence in healthy pouches by the differential effect on lymphocyte proliferation. METHODS: An ex vivo cell culture assay was used in which peripheral blood mononuclear cells or lamina propria mononuclear cells were cultured with sterile sonicates of gut flora from patients with or without pouchitis in the presence of antigen presenting cells. RESULTS: Sonicated pouchitis flora produced a consistent and intense proliferation of the mononuclear cells but that produced by sonicates from healthy pouches was minimal (p = 0.012 or 0.018, peripheral blood or lamina propria mononuclear cells). Preparation of the sonicates with the antibiotic metronidazole abolished their stimulatory ability (p = 0.005, peripheral blood mononuclear cells). In separate assays neither direct addition of metronidazole nor of its hydroxy metabolite affected the mononuclear cells' proliferation with alternative stimuli. CONCLUSIONS: These results strongly support a bacterial aetiology for pouchitis. PMID- 15542519 TI - Clinical and subclinical intestinal inflammation assessed by the mucosal patch technique: studies of mucosal neutrophil and eosinophil activation in inflammatory bowel diseases and irritable bowel syndrome. AB - BACKGROUND AND AIMS: There is a clear need for a rapid, simple, safe, and sensitive method of determining the type and intensity of inflammation in the gut mucosa in clinical practice. In this study, we have evaluated the potential of a new method, the mucosal patch technique, in patients with and without apparent gut inflammation, as assessed by conventional diagnostic procedures. SUBJECTS AND METHODS: The technique tested is based on the idea that inflammatory mediators released from the rectal mucosa can be absorbed by and then extracted from cellulose patches brought into contact with the mucosa by use of an instrument with an inflatable balloon. Measurements were performed in healthy controls (n = 16) and in patients with active (n = 19) and inactive ulcerative colitis (UC, n = 8), collagen colitis (CC, n = 12), coeliac disease (n = 13), and irritable bowel syndrome (IBS, n = 13). RESULTS: Inflammatory mediators from neutrophils (myeloperoxidase (MPO)) and eosinophils (eosinophil cationic protein (ECP)) were increased on average 300- and 10-fold, respectively, in patients with active UC compared with healthy controls and were correlated with the endoscopic score. Patients with inactive UC, CC, coeliac disease, and IBS exhibited no endoscopic signs of inflammation. These patient groups had significantly lower levels of MPO and ECP than the active UC group but showed on average a four- to sevenfold increase in MPO compared with healthy controls. CONCLUSION: The mucosal patch technique was well tolerated by patients and easily applied by the investigator. Pronounced neutrophil and eosinophil involvement in UC was demonstrated. With the high sensitivity of the technique, low degree mucosal neutrophil activation could also be quantified in patients with CC and UC in clinical remission. The finding of increased neutrophil involvement in patients with IBS contributes to the pathophysiological ideas of this disease. PMID- 15542520 TI - Cancer surveillance in longstanding ulcerative colitis: endoscopic appearances help predict cancer risk. AB - BACKGROUND AND AIMS: The risk of colorectal cancer is increased in ulcerative colitis (UC). Patients with UC have diverse colonoscopic appearances. Determining colonoscopic markers for cancer risk could allow patient risk stratification. PATIENTS AND METHODS: Following on from an earlier study which demonstrated a correlation between inflammation severity and neoplasia risk, a case control study was performed to look for colonoscopic markers of colorectal neoplasia risk in UC. Each patient with neoplasia detected between 1988 and 2002 was matched with two non-dysplastic colitic controls. Data were collected on post inflammatory polyps, scarring, strictures, backwash ileitis, a shortened, tubular, or featureless colon, severe inflammation, and normal looking surveillance colonoscopies. RESULTS: Cases (n = 68) and controls (n = 136) were well matched. On univariate analysis, cases were significantly more likely to have post-inflammatory polyps (odds ratio (OR) 2.14 (95% confidence interval 1.24 3.70)), strictures (OR 4.22; 1.08-15.54), shortened colons (OR 10.0; 1.17-85.6), tubular colons (OR 2.03; 1.00-4.08), or segments of severe inflammation (OR 3.38; 1.41-10.13), and less likely to have had a macroscopically normal looking colonoscopy (OR 0.40; 0.21-0.74). After multivariate analysis, a macroscopically normal looking colonoscopy (OR 0.38; 0.19-0.73), post-inflammatory polyps (2.29; 1.28-4.11), and strictures (4.62; 1.03-20.8) remained significant. The five year risk of colorectal cancer following a normal looking colonoscopy was no different from that of matched general population controls. CONCLUSIONS: Macroscopic colonoscopic features help predict neoplasia risk in UC. Features of previous/ongoing inflammation signify an increased risk. A macroscopically normal looking colonoscopy returns the cancer risk to that of the general population: it should be possible to reduce surveillance frequency to five years in this cohort. PMID- 15542521 TI - Increased epithelial uptake of protein antigens in the ileum of Crohn's disease mediated by tumour necrosis factor alpha. AB - BACKGROUND AND AIMS: The exact nature of the epithelial barrier defect in Crohn's disease remains to be elucidated. Previously we showed increased permeability to proteins in ileal Crohn's disease. Our aims were to study if this barrier defect (a) involves endocytotic uptake of antigens and (b) is related to low grade inflammation not detectable by histology. METHODS: Macroscopically normal segments of distal ileum of Crohn's disease patients (n = 10) were subgrouped into non-inflamed (histologically unaffected) and slightly inflamed tissues and studied in Ussing chambers, with normal ileal specimens from colon cancer patients (n = 9) as controls. Endocytotic uptake into enterocytes of the protein antigen horseradish peroxidase was assessed by measuring the area of horseradish peroxidase containing endosomes in electron photomicrographs. Mucosal tumour necrosis factor alpha (TNF-alpha) mRNA was quantified using real time polymerase chain reaction. For comparison, the effects of low doses of TNF-alpha on endosomal uptake of horseradish peroxidase were studied in cultured T84 cells grown on filter supports. RESULTS: The area of horseradish peroxidase containing endosomes was increased (p<0.001) in enterocytes of non-inflamed ileum of Crohn's disease (2.8 (0.7) mum(2)/300 mum(2)) compared with control ileum (0.6 (0.06)). In non-inflamed mucosa, a significant association between endosomal uptake and mucosal expression of TNF-alpha mRNA (p = 0.03) was found. Low concentrations of TNF-alpha (0.25-1.0 ng/ml) enhanced the endosomal uptake of horseradish peroxidase in polarised T84 cells, without affecting transepithelial electrical resistance. CONCLUSIONS: Our findings suggest increased endosomal uptake of antigens in ileal Crohn's disease that may be mediated by TNF-alpha. These data highlight the transcellular route of antigen uptake in barrier dysfunction and implicate the interaction between epithelial cells and the innate immune system in the development of mucosal inflammation. PMID- 15542522 TI - An unusual case of colitis. PMID- 15542523 TI - Effects of folylpolyglutamate synthetase modulation on chemosensitivity of colon cancer cells to 5-fluorouracil and methotrexate. AB - BACKGROUND: Folylpoly-gamma-glutamate synthetase (FPGS) converts intracellular folates and antifolates (for example, methotrexate (MTX)) to polyglutamates. Polyglutamylated folates and antifolates are retained in cells longer and are better substrates than their monoglutamate counterparts for enzymes involved in one carbon transfer. Polyglutamylation of intracellular 5,10 methylenetetrahydrofolate may also enhance the cytotoxicity of 5-fluorouracil (5 FU) by allowing more efficient formation and stabilisation of the inhibitory ternary complex involving thymidylate synthase and a 5-FU metabolite. AIM: We investigated the effects of FPGS modulation on the chemosensitivity of colon cancer cells to 5-FU and MTX. METHODS: Human HCT116 colon cancer cells were stably transfected with the sense or antisense FPGS cDNA or blank (control). FPGS protein expression and enzyme activity, growth rate, intracellular folate content and composition, and in vitro chemosensitivity to 5-FU and MTX were determined. RESULTS: Compared with cells expressing endogenous FPGS, those overexpressing FPGS had significantly faster growth rates and higher concentrations of total folate and long chain folate polyglutamates while antisense FPGS inhibition produced opposite results. FPGS overexpression significantly enhanced, whereas FPGS inhibition decreased, chemosensitivity to 5-FU. No significant difference in chemosensitivity to MTX was observed. CONCLUSIONS: These data provide functional evidence that FPGS overexpression and inhibition modulate chemosensitivity of colon cancer cells to 5-FU by altering intracellular folate polyglutamylation, providing proof of principle. Thus FPGS status may be an important predictor of chemosensitivity of colon cancer cells to 5-FU based chemotherapy, and FPGS gene transfer may increase the sensitivity of colon cancer cells to 5-FU-based chemotherapy. PMID- 15542524 TI - Evidence for genetic predisposition to desmoid tumours in familial adenomatous polyposis independent of the germline APC mutation. AB - BACKGROUND: Many patients with familial adenomatous polyposis (FAP) die from desmoid tumours which can arise spontaneously but often appear to be surgically induced by prophylactic colectomy. FAP results from germline adenomatous polyposis coli (APC) gene mutations and desmoids arise following biallelic APC mutation, with one change usually occurring distal to the second beta-catenin binding/degradation repeat of the gene (3' to codon 1399). We have suggested that because families with germline mutations in this region already have the requisite change, they are more likely to develop desmoids. However, there are families with 5' germline mutations where desmoids are common. PATIENTS AND METHODS: We examined desmoid risk dependent on germline APC mutation, sex, history of abdominal surgery, and family history in FAP patients from the St Mark's Hospital Polyposis Registry. RESULTS: Overall desmoid prevalence was 15%. Desmoids tended to cluster in susceptible individuals, irrespective of the germline APC mutation. Independent predictors of increased desmoid risk were: germline mutation distal to codon 1399; any family history of disease; and a strong family history of desmoids. A family history of multiple desmoids (>1) increased an individual's own risk of multiplicity. Females had twice the odds of developing desmoids compared with males. There was no significant interaction between any of the three explanatory variables. CONCLUSIONS: Our results indicate the influence of unknown genetic factors independent of APC in susceptibility to desmoid tumours in FAP. The data have implications in terms of clinical management of FAP patients and assessing the balance between chemoprevention and prophylactic colectomy. PMID- 15542525 TI - Bile secretory function in the obese Zucker rat: evidence of cholestasis and altered canalicular transport function. AB - BACKGROUND: Obese Zucker rats (ZR) have been used as an experimental model for non-alcoholic fatty liver disease and are particularly susceptible to various types of liver injury. Bile secretory function has not been assessed in ZR. AIM: To study bile secretion and expression of the main hepatobiliary transporters in ZR. METHODS: Bile flow and biliary secretion of lipids and glutathione were determined in eight and 14 week old obese ZR and their lean controls. Protein mass and mRNA of the Na(+)/taurocholate cotransporting polypeptide (Ntcp), the bile salt export pump (Bsep), and the multidrug resistant associated protein 2 (Mrp2) were assessed by western and northern blot, respectively. The effects of administration of a tumour necrosis factor alpha inactivator (etanercept) and an insulin sensitiser (rosiglitazone) were assessed in obese ZR while leptin was given to non-obese rats to study its effect on Mrp2 expression. RESULTS: ZR exhibited increased body weight and hyperlipidaemia. Only 14 week old obese ZR has fatty liver. Decreased bile flow and biliary lipid and glutathione secretion as well as reduced hepatic transport of both taurocholate and bromosulphthalein were found in obese ZR. Hepatic Mrp2 protein mass was markedly reduced (-70%) in obese rats while Ntcp and Bsep protein levels were similar to lean rats. Downregulation of Mrp2 seems to involve both transcriptional and post transcriptional mechanisms probably related to insulin and leptin resistance. CONCLUSIONS: Obese ZR exhibit an impaired bile secretory function with significant functional and molecular alterations consistent with mild cholestasis. A defective hepatobiliary transport capacity may be a contributory factor in rendering the obese ZR more susceptible to liver injury. PMID- 15542526 TI - Tezosentan, an endothelin receptor antagonist, limits liver injury in endotoxin challenged cirrhotic rats. AB - BACKGROUND/AIMS: Lipopolysaccharide (LPS) induces liver injury which is associated with upregulated endothelin (ET)-1 production. The aim of this study was to investigate the effects of tezosentan, a non-selective ETA and ETB receptor antagonist, in LPS challenged rats with cirrhosis. METHODS: Rats with cirrhosis received LPS and then tezosentan or placebo one hour later. Four hours after LPS administration, rats were killed to measure serum transaminase activity and plasma tumour necrosis factor alpha (TNF-alpha) levels. Hepatic inducible nitric oxide synthase (iNOS), myeloperoxidase (MPO), a marker of neutrophil infiltration, and cyclooxygenase (COX)-2 expression were also measured. RESULTS: LPS administration significantly decreased arterial pressure and significantly increased plasma endothelin levels. Following LPS and tezosentan administration, serum aspartate aminotransferase and alanine aminotransferase activities were similar to those in the control group while they were increased by more than 700% with LPS alone. Plasma TNF-alpha levels were significantly lower in rats receiving LPS and tezosentan (182 (38) pg/ml) compared with those receiving LPS alone (821 (212) pg/ml). Tezosentan significantly decreased hepatic MPO activity and hepatic neutrophils but had no effect on LPS induced iNOS or COX-2. Survival rate was significantly higher in rats receiving LPS plus tezosentan (80%) than in rats receiving LPS alone (50%). CONCLUSION: In LPS challenged cirrhotic rats, tezosentan administration prevents LPS induced liver injury by decreasing intrahepatic neutrophil infiltration. In addition, tezosentan increases survival in these rats. PMID- 15542527 TI - Influence of biliary cirrhosis on the detoxification and elimination of a food derived carcinogen. AB - BACKGROUND AND AIMS: The liver is the central organ for the detoxification of numerous xenobiotics, including carcinogens. We studied the influence of cholestasis and biliary cirrhosis on the detoxification, elimination, and tissue distribution of a model compound and food derived carcinogen, 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine (PhIP). METHODS: Wistar rats were injected with (14)C-PhIP into the portal vein one or six weeks after common bile duct ligation (CBDL). Bile flow was reconstituted, bile and urine were collected over 120 minutes, and metabolites were analysed using high performance liquid chromatograpy. Total tissue radioactivity levels in several organs as well as tissue bound (ethanol insoluble tissue fraction) radioactivity levels were determined. RESULTS: Significant downregulation of the transport proteins multidrug resistance associated protein 2 and breast cancer resistance protein was observed in biliary cirrhosis. Biliary excretion of radioactivity was significantly reduced in cholestasis and biliary cirrhosis compared with controls (15 (2.9)% and 3.2 (1)% of the dose v 36.5 (2)%, respectively). Phase II metabolism was severely reduced in cirrhotic rats, resulting in a twofold increase in tissue radioactivity levels in the liver, kidney, and colon. Biliary cirrhosis increased tissue binding of reactive metabolites, as expressed in cpm/100 mg tissue in the liver and the colon (3267 (1218) v 1191 (429) in the liver, 3044 (1913) v 453 (253) in the colon). CONCLUSIONS: Biliary cirrhosis induced by CBDL causes impaired metabolism and elimination of PhIP, and leads to higher tissue levels of potentially genotoxic metabolites in the liver and colon of rats. These data may explain the increased incidence of hepatic and extrahepatic cancers in cholestasis and liver cirrhosis. PMID- 15542528 TI - Endoscopic retreatment of recurrent choledocholithiasis after sphincterotomy. AB - BACKGROUND: Endoscopic sphincterotomy (ES) carries a substantial risk of recurrent choledocholithiasis but retreatment with endoscopic retrograde cholangiopancreatography (ERCP) is safe and feasible. However, long term results of repeat ERCP and risk factors for late complications are largely unknown. AIMS: To investigate the long term outcome of repeat ERCP for recurrent bile duct stones after ES and to identify risk factors predicting late choledochal complications. METHODS: Eighty four patients underwent repeat ERCP, combined with ES in 69, for post-ES recurrent choledocholithiasis. Long term outcomes of repeat ERCP were retrospectively investigated and factors predicting late complications were assessed by multivariate analysis. RESULTS: Complete stone clearance was achieved in all patients. Forty nine patients had no visible evidence of prior sphincterotomy. Two patients experienced early complications. During a follow up period of 2.2-26.0 years (median 10.9 years), 31 patients (37%) developed late complications, including stone recurrence (n = 26), acute acalculous cholangitis(n = 4), and acute cholecystitis (n = 1). There were neither biliary malignancies nor deaths attributable to biliary disease. Multivariate analysis identified three independent risk factors for choledochal complications: interval between initial ES and repeat ERCP < or =5 years, bile duct diameter > or =15 mm, and periampullary diverticulum. Choledochal complications were successfully treated with repeat ERCP in 29 patients. CONCLUSIONS: Choledochal complications after repeat ERCP are relatively frequent but are endoscopically manageable. Careful follow up is necessary, particularly for patients with a dilated bile duct, periampullary diverticulum, or early recurrence. Repeat ERCP is a reasonable treatment even for recurrent choledocholithiasis after ES. PMID- 15542529 TI - Use of microsatellite marker loss of heterozygosity in accurate diagnosis of pancreaticobiliary malignancy from brush cytology samples. AB - BACKGROUND: Brush cytology of biliary strictures to diagnose pancreaticobiliary malignancy suffers from poor sensitivity. AIM: To improve the diagnostic yield of pancreaticobiliary brush cytology through analysis of tumour suppressor gene linked microsatellite marker loss of heterozygosity (LOH) and k-ras codon 12 mutation detection. METHODS: Twenty six patients with biliary strictures underwent endoscopic retrograde cholangiography with brush cytology. A panel of 12 polymorphic microsatellite markers linked to six tumour suppressor genes was developed. Genomic DNA from cell clusters acquired from brush cytology specimens and microdissected surgical malignant and normal tissue underwent polymerase chain amplification reaction (PCR). PCR products were compared for LOH and k-ras codon 12 mutations. RESULTS: Seventeen patients were confirmed to have pancreaticobiliary adenocarcinoma. Nine patients had benign strictures (eight proven surgically, one by follow up). Cytomorphological interpretation was positive for malignancy (n = 8), indeterminate (n = 10), and negative for malignancy (n = 8). Selected malignant appearing cytological cell clusters and microdissected histological samples from cancer showed abundant LOH characteristic of malignancy while brushings from nine cases without cancer carried no LOH (p<0.001). LOH and k-ras mutations profile of the cytological specimens was almost always concordant with the tissue samples. Presence of k-ras mutation predicted malignancy of pancreatic origin (p<0.001). CONCLUSION: LOH and k-ras codon 12 mutation analysis of PCR amplified DNA from biliary brush cytology discriminates reactive from malignant cells, with 100% sensitivity, specificity, and accuracy. Minor variations in LOH in brushings and in different sites within the same tumour likely reflect intratumoral mutational heterogeneity during clonal expansion of pre- and neoplastic lineages. PMID- 15542530 TI - Role of video endoscopy in managing small bowel disease. PMID- 15542531 TI - Should we remove all lesions at colonoscopy? PMID- 15542532 TI - Ultrastructural changes in enterocytes in subjects with Hashimoto's thyroiditis. PMID- 15542533 TI - Liver fibrosis: natural history may be affected by the biopsy sample. PMID- 15542535 TI - Methodology for estimating regional and global trends of child malnutrition. AB - BACKGROUND: Child malnutrition is an important indicator for monitoring progress towards the Millennium Development Goals (MDG). This paper describes the methodology developed by the World Health Organization (WHO) to derive global and regional trends of child stunting and underweight, and reports trends in prevalence and numbers affected for 1990-2005. METHODS: National prevalence data from 139 countries were extracted from the WHO Global Database on Child Growth and Malnutrition. A total of 419 and 388 survey data points were available for underweight and stunting, respectively. To estimate trends we used linear mixed effect models allowing for random effects at country level and for heterogeneous covariance structures. One model was fitted for each United Nation's region using the logit transform of the prevalence and results back-transformed to the original scale. Best models were selected based on explicit statistical and graphical criteria. RESULTS: During 1990-2000 global stunting and underweight prevalences declined from 34% to 27% and 27% to 22%, respectively. Large declines were achieved in Eastern and South-eastern Asia, while South-central Asia continued to suffer very high levels of malnutrition. Substantial improvements were also made in Latin America and the Caribbean, whereas in Africa numbers of stunted and underweight children increased from 40 to 45, and 25 to 31 million, respectively. CONCLUSION: Linear mixed-effect models made best use of all available information. Trends are uneven across regions, with some showing a need for more concerted and efficient interventions to meet the MDG of reducing levels of child malnutrition by half between 1990 and 2015. PMID- 15542534 TI - Overdiagnosis of malaria in patients with severe febrile illness in Tanzania: a prospective study. AB - OBJECTIVE: To study the diagnosis and outcomes in people admitted to hospital with a diagnosis of severe malaria in areas with differing intensities of malaria transmission. DESIGN: Prospective observational study of children and adults over the course a year. SETTING: 10 hospitals in north east Tanzania. PARTICIPANTS: 17,313 patients were admitted to hospital; of these 4474 (2851 children aged under 5 years) fulfilled criteria for severe disease. MAIN OUTCOME MEASURE: Details of the treatment given and outcome. Altitudes of residence (a proxy for transmission intensity) measured with a global positioning system. RESULTS: Blood film microscopy showed that 2062 (46.1%) of people treated for malaria had Plasmodium falciparum (slide positive). The proportion of slide positive cases fell with increasing age and increasing altitude of residence. Among 1086 patients aged > or = 5 years who lived above 600 metres, only 338 (31.1%) were slide positive, while in children < 5 years living in areas of intense transmission (< 600 metres) most (958/1392, 68.8%) were slide positive. Among 2375 people who were slide negative, 1571 (66.1%) were not treated with antibiotics and of those, 120 (7.6%) died. The case fatality in slide negative patients was higher (292/2412, 12.1%) than for slide positive patients (142/2062, 6.9%) (P < 0.001). Respiratory distress and altered consciousness were the strongest predictors of mortality in slide positive and slide negative patients and in adults as well as children. CONCLUSIONS: In Tanzania, malaria is commonly overdiagnosed in people presenting with severe febrile illness, especially in those living in areas with low to moderate transmission and in adults. This is associated with a failure to treat alternative causes of severe infection. Diagnosis needs to be improved and syndromic treatment considered. Routine hospital data may overestimate mortality from malaria by over twofold. PMID- 15542536 TI - Exhaled nitric oxide before and after cardiac surgery with cardiopulmonary bypass -response to acetylcholine and nitroglycerin. AB - BACKGROUND: Pulmonary endothelial dysfunction may occur after ischaemia reperfusion injury and can be revealed as a reduced vasodilatory response upon administration of acetylcholine (ACh). ACh also releases the endothelium-derived vasodilator nitric oxide but direct measurements of this gas are difficult to perform in vivo. We wanted to study the effects of i.v. administration of ACh and the endothelium-independent vasodilator nitroglycerin on exhaled nitric oxide in relation to pulmonary endothelial dysfunction after open-heart surgery and cardiopulmonary bypass (CPB). METHODS: Basal exhaled nitric oxide and the response in exhaled nitric oxide to i.v. injections of ACh and nitroglycerin were measured with chemiluminescence in 10 patients before and after open-heart surgery. RESULTS: Exhaled nitric oxide decreased significantly after CPB. I.V. bolus injections of ACh induced a reproducible and dose-dependent increase in exhaled nitric oxide that was unaltered after CPB. In contrast, the increase in exhaled nitric oxide evoked by nitroglycerin was attenuated after CPB. The response in pulmonary vascular resistance index (PVRI) to an infusion of ACh decreased after CPB, indicating endothelial dysfunction. The decrease in PVRI response to ACh correlated to the duration of CPB. CONCLUSIONS: Interestingly, pulmonary vascular dysfunction after CPB was accompanied by a reduction in the exhaled nitric oxide response to nitroglycerin and lower levels of basal exhaled nitric oxide. The ACh-induced responses in exhaled nitric oxide were unchanged, which could indicate nitric oxide-independent mechanisms behind the endothelial dysfunction in this study. The possibility of using exhaled nitric oxide dynamics to investigate pulmonary endothelial dysfunction merits further studies. PMID- 15542537 TI - Role of protein kinase C-epsilon (PKCepsilon) in isoflurane-induced cardioprotection. AB - BACKGROUND: Volatile anaesthetics precondition the heart against infarction, an effect partly mediated by activation of the epsilon isoform of protein kinase C (PKCepsilon). We investigated whether cardioprotection by activation of PKCepsilon depends on the isoflurane concentration. METHODS: Anaesthetized rats underwent 25 min of coronary artery occlusion followed by 120 min of reperfusion and were randomly assigned to the following groups (n=10 in each group): isoflurane preconditioning induced by 15 min administration of 0.4 minimal alveolar concentration (MAC) (0.4MAC), 1 MAC (1MAC) or 1.75 MAC (1.75MAC) followed by 10 min washout before ischaemia. Each protocol was repeated in the presence of the PKC inhibitor staurosporine (10 microg kg(-1)): 0.4MAC+S, 1MAC+S and 1.75MAC+S. Controls were untreated (CON) and additional hearts received staurosporine without isoflurane (S). In a second set of experiments (n=6 in each group) hearts were excised before the infarct inducing ischaemia, and phosphorylation and translocation of PKCepsilon were determined by western blot analysis. RESULTS: Isoflurane reduced infarct size from a mean of 61(SEM 2)% of the area at risk in controls to 20(1)% (0.4MAC), 26(3)% (1MAC) and 30(1)% (1.75MAC) (all P<0.01 vs CON or S). This protection was partially reversed by administration of staurosporine in the 0.4MAC+S group (30[2]%; P<0.05 vs 0.4MAC) group, but not after administration of 1 MAC or 1.75 MAC isoflurane (26[2]% and 31[2]%, respectively). Thus 0.4MAC increased PKCepsilon phosphorylation, and this effect was blocked by staurosporine. Higher concentrations of isoflurane did not change PKCepsilon phosphorylation. PKCepsilon was translocated to the membrane fraction after administration of 0.4 MAC isoflurane, but not after 1.0 or 1.75 MAC. CONCLUSIONS: Although isoflurane preconditioning resulted in a reduction in infarct size at all concentrations used, the protection was mediated by phosphorylation and translocation of PKCepsilon only at 0.4 MAC. PMID- 15542538 TI - Effect of timing on the response to postal questionnaires concerning satisfaction with anaesthesia care. AB - BACKGROUND: There is little information on the effect of time on the assessment by the patient of quality of anaesthesia care. This study compared the patient's assessment of anaesthesia care after three different periods of time following discharge from hospital. Materials. Three groups of patients were assigned to receive a standardized, validated psychometric questionnaire either 1, 5, or 9 weeks after discharge from hospital. We measured response rate and the total mean problem score of six dimensions. RESULTS: Groups 1, 2, and 3 received 748, 743, and 723 questionnaires, respectively. The response rates including one reminder were 67.3 (95% confidence interval [CI] 63.9-70.6%), 64.5% (CI 61.1-67.9%), and 58.9% (CI 55.5-62.4%), respectively (Group 1 vs Group 3, P<0.001, and Group 2 vs Group 3, P<0.05). The total mean problem scores were not significantly different with 17 (CI 1.4%), 17 (CI 1.4%), and 15% (CI 1.3%), respectively. In two out of six dimensions ('Continuity of personal care by anaesthetist' and 'Nursing care in recovery room') significantly less problems were reported after 9 weeks. The other dimensions of the questionnaire showed no consistent differences between groups. CONCLUSIONS: The response rate is significantly lower at 9 weeks compared with 1 and 5 weeks after discharge. The total mean problem score remains unchanged but certain fields show fewer problems after 9 weeks compared with 1 and 5 weeks. Questionnaires on patient satisfaction with anaesthesia care should be sent within 5 weeks of discharge. PMID- 15542539 TI - Interstitial muscle concentrations of rocuronium under steady-state conditions in anaesthetized dogs: actual versus predicted values. AB - INTRODUCTION: The objective of this study was to compare rocuronium effect (C(e)) and peripheral (C(2)) compartment concentrations predicted by pharmacokinetic pharmacodynamic (PK-PD) modelling with those measured in plasma (C(p)) and in the interstitial fluid of muscle tissue (C(ISF,u)) by microdialysis in anaesthetized dogs. METHODS: After approval by the Animal Care Committee, eight adult male dogs with a body weight ranging from 7 to 18 kg were anaesthetized with pentobarbital. Each dog received a 2-min rocuronium infusion of 0.15 mg kg(-1) min(-1) followed by a 118-min infusion of 60 microg kg(-1) min(-1) via the right jugular vein. Arteriovenous gradient across the hindlimb was measured at 40, 60, 100 and 120 min. Three microdialysis samples were collected at 40-min intervals. Once the infusion stopped, arterial samples were collected every 2 min for the first 10 min and every 20 min for the next 120 min. Neuromuscular function was monitored using train-of-four stimulation until full recovery. Dogs were then killed and a biopsy of muscle tissue was performed (C(m)). RESULTS: At steady state, the mean C(ISF,u) value was 1353 ng ml(-1). After correction for the unbound fraction in plasma, the mean C(e,corr) and C(2,corr) were 1681 and 1481 ng ml(-1), respectively. At the terminal sampling point, C(m) was 10-fold higher than C(p). CONCLUSION: Unbound concentration of rocuronium measured in the muscle interstitial fluid under steady-state conditions confirms that parametric PK-PD modelling gives reliable estimates of effect site concentrations. Rocuronium accumulates in muscle tissue, probably by non-specific protein binding in the interstitial space. PMID- 15542540 TI - Improved design of an antigen with enhanced specificity for the broadly HIV neutralizing antibody b12. AB - In an attempt to design immunogens that elicit broadly HIV-neutralizing antibodies, we recently engineered monomeric HIV-1 gp120 to bind preferentially b12, a broadly neutralizing antibody to the CD4-binding site (CD4bs) on gp120, by mutating four central residues in the CD4bs to alanine and introducing extra N glycosylation sites potentially to mask unwanted B-cell epitopes. Despite the favorable antigenicity of this mutant, it harbors two potential caveats that may limit its effectiveness to elicit b12-like antibodies: (i) b12-binding affinity is reduced relative to wild-type gp120 and (ii) binding of some non-neutralizing antibodies to the N-terminal C1 region of gp120 is still observed. Here, we sought to correct these potential limitations. By reverting one of the added N glycosylation sites on the gp120 core, b12 binding was improved without affecting the epitope-masking properties of the original mutant. Furthermore, truncation of the gp120 N-terminus eliminated binding of the anti-C1 antibodies. Finally, based on the binding profiles of additional non-neutralizing antibodies tested here, further N-glycosylation sites were incorporated to mask their corresponding epitopes. The resulting hyperglycosylated gp120 variants bind b12 and another broadly neutralizing antibody, 2G12, with apparent affinities approaching that of wild-type gp120, but do not bind 21 non- or weakly neutralizing antibodies to seven different epitopes on gp120. These hyperglycosylated variants expand our panel of glycoengineered gp120s that are currently being evaluated for their ability to elicit broadly neutralizing antibodies. PMID- 15542541 TI - alpha-SNAP and NSF are required in a priming step during the human sperm acrosome reaction. AB - The acrosome is a membrane-limited granule that overlies the nucleus of the mature spermatozoon. In response to physiological or pharmacological stimuli it undergoes a special type of Ca2+-dependent exocytosis termed the acrosome reaction (AR), which is an absolute prerequisite for fertilization. Aided by a streptolysin-O permeabilization protocol developed in our laboratory, we have previously demonstrated requirements for Rab3A, N-ethylmaleimide-sensitive factor (NSF), several soluble NSF-attachment protein receptor (SNARE) proteins, and synaptotagmin VI in the human sperm AR. Here, we show that alpha-soluble NSF attachment protein (alpha-SNAP), a protein essential for most fusion events through its interaction with NSF and the SNARE complex, exhibits a direct role in the AR. First, the presence of alpha-SNAP is demonstrated by the Western blot of human sperm protein extracts. Immunostaining experiments reveal an acrosomal localization for this protein. Second, the Ca2+ and Rab3A-triggered ARs are inhibited by anti-alpha-SNAP antibodies. Third, bacterially expressed alpha-SNAP abolishes exocytosis in a fashion that depends on its interaction with NSF. Fourth, we show a requirement for alpha-SNAP/NSF in a prefusion step early in the exocytotic pathway, after the tethering of the acrosome to the plasma membrane and before the efflux of intra-acrosomal Ca2+. These results suggest a key role for alpha-SNAP/NSF in the AR, and strengthen our understanding of the molecular players involved in the vesicle-to-plasma membrane fusion taking place during exocytosis. PMID- 15542542 TI - Burn injury induces a change in T cell homeostasis affecting preferentially CD4+ T cells. AB - Burn injuries are known to be associated with altered immune functions, resulting in decreased resistance to subsequent infection. In the present study, we determined the in vivo changes in T cell homeostasis following burn injury. Two groups of mice were used: a sham-burn group receiving buprenorphine as an analgesic and a burn group receiving buprenorphine and subjected to burn injury on 20% of the total body surface area. Results showed an important decrease in splenocytes following burn injury. This decrease persisted for 5 days and was followed, at day 10, by a 63% increase in number of cells. In vivo cell proliferation, as determined by the incorporation of 5-bromo-2'-dexoxyuridine, showed a significant increase of cycling splenocytes between days 2 and 10 after burn injury. The percentage of CD4+ and CD8+ T cells in the spleen was altered for 10 days after thermal injury. Analysis of naive (CD62Lhigh CD44low) and effector/memory (CD62Llow CD44high) T cells showed a percent decrease, independent of the expression of CD4 or CD8 molecules. However, early activation markers, such as CD69+, were expressed only on CD4+ T cells after a number of days following injury. Even with an activated phenotype, 10 days post-burn injury, CD4+ naive T cells significantly increased spontaneous apoptosis, detected by using a fluorescent DNA-binding agent 7-amino-actinomycin D. CD8+ T lymphocytes did not express early activation markers and were more resistant to apoptosis. Using purified T cells, we have shown unresponsiveness at day 10. Overall, these results demonstrate that mechanisms of T cell homeostasis were perturbed following burn injury. However, after 10 days, this perturbation persisted only in CD4+ T cells. PMID- 15542543 TI - Properties and expression of human tartrate-resistant acid phosphatase isoform 5a by monocyte-derived cells. AB - Human serum tartrate-resistant acid phosphatase exists as two enzyme isoforms (TRACP 5a and 5b), derived by differential, post-translational processing of a common gene product. Serum TRACP 5b is from bone-resorbing osteoclasts (OC) and becomes elevated in diseases of increased bone resorption. TRACP 5a is secreted by macrophages (MPhi) and dendritic cells (DC) and is increased in many patients with rheumatoid arthritis. Our purpose was to fully characterize the properties of human TRACP isoforms and to produce an antibody specific to TRACP 5a for use as a biomarker in chronic inflammatory diseases. Partially purified, natural serum TRACP isoforms and recombinant TRACP 5a (rTRACP 5a) were compared with respect to specific activity and subunit structure and presence of sialic acid. Mice were immunized with rTRACP 5a, and resulting hybridomas were screened for monoclonal antibody to serum TRACP 5a. One antibody, 220, was tested for its epitope specificity and use in various immunological techniques. rTRACP 5a had properties identical to serum TRACP 5a. Antibody 220 was specific for the trypsin sensitive epitope in the loop peptide, present only in TRACP 5a. Antibody 220 was effective for specific immunoprecipitation, immunoassay, and immunoblot of TRACP 5a. Intact TRACP was present in MPhi, DC, and OC. TRACP 5a was the predominant isoform secreted by MPhi and DC, whereas TRACP 5b was the predominant isoform secreted by OC. TRACP isoforms 5a and 5b may have different functions inside and outside of monocyte-derived cells. Antibody 220 is an important resource for studies of the biosynthetic relationship among TRACP isoforms and of the significance of serum TRACP 5a as a marker in diseases of bone metabolism and inflammation. PMID- 15542544 TI - Hyperoxia alters the expression and phosphorylation of multiple factors regulating translation initiation. AB - Hyperoxia is cytotoxic and depresses many cellular metabolic functions including protein synthesis. Translational control is exerted primarily during initiation by two mechanisms: 1) through inhibition of translation initiation complex formation via sequestration of the cap-binding protein, eukaryotic initiation factor (eIF) 4E, with inhibitory 4E-binding proteins (4E-BP); and 2) by prevention of eIF2-GTP-tRNA(i)(Met) formation and eIF2B activity by phosphorylated eIF2alpha. In this report, exposure of human lung fibroblasts to 95% O2 decreased the incorporation of thymidine into DNA at 6 h and the incorporation of leucine into protein beginning at 12 h. The reductions in DNA and protein synthesis were accompanied by increased phosphorylation of eIF4E protein and reduced phosphorylation of 4E-BP1. At 24 h, hyperoxia shifted 4E-BP1 phosphorylation to lesser-phosphorylated isoforms, increased eIF4E expression, and increased the association of eIF4E with 4E-BP1. Although hyperoxia did not change eIF2alpha expression, it increased its phosphorylation at Ser51, but not until 48 h. In addition, the activation of eIF2alpha was not accompanied by the formation of stress granules. These findings suggest that hyperoxia diminishes protein synthesis by increasing eIF4E phosphorylation and enhancing the affinity of 4E-BP1 for eIF4E. PMID- 15542545 TI - Both cAMP and cGMP are required for maximal ciliary beat stimulation in a cell free model of bovine ciliary axonemes. AB - Previously, we have shown that the ATPase-dependent motion of cilia in bovine bronchial epithelial cells (BBEC) can be regulated through the cyclic nucleotides, cAMP via the cAMP-dependent protein kinase (PKA) and cGMP via the cGMP-dependent protein kinase (PKG). Both cyclic nucleotides cause an increase in cilia beat frequency (CBF). We hypothesized that cAMP and cGMP may act directly at the level of the ciliary axoneme in BBEC. To examine this, we employed a novel cell-free system utilizing detergent-extracted axonemes. Axoneme movement was whole-field analyzed digitally with the Sisson-Ammons video analysis system. A suspension of extracted axonemes remains motionless until the addition of 1 mM ATP that establishes a baseline CBF similar to that seen when analyzing intact ciliated BBEC. Adding 10 microM cAMP or 10 microM cGMP increases CBF beyond the established ATP baseline. However, the cyclic nucleotides did not stimulate CBF in the absence of ATP. Therefore, the combination of cAMP and cGMP augments ATP driven CBF increases at the level of isolated axoneme. PMID- 15542546 TI - Self-consistent proteomic field theory of stochastic gene switches. AB - We present a self-consistent field approximation approach to the problem of the genetic switch composed of two mutually repressing/activating genes. The protein and DNA state dynamics are treated stochastically and on an equal footing. In this approach the mean influence of the proteomic cloud created by one gene on the action of another is self-consistently computed. Within this approximation a broad range of stochastic genetic switches may be solved exactly in terms of finding the probability distribution and its moments. A much larger class of problems, such as genetic networks and cascades, also remain exactly solvable with this approximation. We discuss, in depth, certain specific types of basic switches used by biological systems and compare their behavior to the expectation for a deterministic switch. PMID- 15542547 TI - Distance-restrained docking of rifampicin and rifamycin SV to RNA polymerase using systematic FRET measurements: developing benchmarks of model quality and reliability. AB - We are developing distance-restrained docking strategies for modeling macromolecular complexes that combine available high-resolution structures of the components and intercomponent distance restraints derived from systematic fluorescence resonance energy transfer (FRET) measurements. In this article, we consider the problem of docking small-molecule ligands within macromolecular complexes. Using simulated FRET data, we have generated a series of benchmarks that permit estimation of model accuracy based on the quantity and quality of FRET-derived distance restraints, including the number, random error, systematic error, distance distribution, and radial distribution of FRET-derived distance restraints. We find that expected model accuracy is 10 A or better for models based on: i), > or =20 restraints with up to 15% random error and no systematic error, or ii), > or =20 restraints with up to 15% random error, up to 10% systematic error, and a symmetric radial distribution of restraints. Model accuracies can be improved to 5 A or better by increasing the number of restraints to > or =40 and/or by optimizing the distance distribution of restraints. Using experimental FRET data, we have defined the positions of the binding sites within bacterial RNA polymerase of the small-molecule inhibitors rifampicin (Rif) and rifamycin SV (Rif SV). The inferred binding sites for Rif and Rif SV were located with accuracies of, respectively, 7 and 10 A relative to the crystallographically defined binding site for Rif. These accuracies agree with expectations from the benchmark simulations and suffice to indicate that the binding sites for Rif and Rif SV are located within the RNA polymerase active center cleft, overlapping the binding site for the RNA-DNA hybrid. PMID- 15542548 TI - DNA ejection from bacteriophage T5: analysis of the kinetics and energetics. AB - DNA ejection from bacteriophage T5 can be passively driven in vitro by the interaction with its specific host receptor. Light scattering was used to determine the physical parameters associated with this process. By studying the ejection kinetics at different temperatures, we demonstrate that an activation energy of the order of 70 k(B)T must be overcome to allow the complete DNA ejection. A complex shape of the kinetics was found whatever the temperature. This shape may be actually understood using a phenomenological model based on a multistep process. Passing from one stage to another requires the mentioned thermal activation of pressurized DNA inside the capsids. Both effects contribute to shorten or to lengthen the pause time between the different stages explaining why the T5 DNA ejection is so slow compared to other types of phage. PMID- 15542549 TI - Elucidating protein thermodynamics from the three-dimensional structure of the native state using network rigidity. AB - Given the three-dimensional structure of a protein, its thermodynamic properties are calculated using a recently introduced distance constraint model (DCM) within a mean-field treatment. The DCM is constructed from a free energy decomposition that partitions microscopic interactions into a variety of constraint types, i.e., covalent bonds, salt-bridges, hydrogen-bonds, and torsional-forces, each associated with an enthalpy and entropy contribution. A Gibbs ensemble of accessible microstates is defined by a set of topologically distinct mechanical frameworks generated by perturbing away from the native constraint topology. The total enthalpy of a given framework is calculated as a linear sum of enthalpy components over all constraints present. Total entropy is generally a nonadditive property of free energy decompositions. Here, we calculate total entropy as a linear sum of entropy components over a set of independent constraints determined by a graph algorithm that builds up a mechanical framework one constraint at a time, placing constraints with lower entropy before those with greater entropy. This procedure provides a natural mechanism for enthalpy-entropy compensation. A minimal DCM with five phenomenological parameters is found to capture the essential physics relating thermodynamic response to network rigidity. Moreover, two parameters are fixed by simultaneously fitting to heat capacity curves for histidine binding protein and ubiquitin at five different pH conditions. The three free parameter DCM provides a quantitative characterization of conformational flexibility consistent with thermodynamic stability. It is found that native hydrogen bond topology provides a key signature in governing molecular cooperativity and the folding-unfolding transition. PMID- 15542550 TI - Line tension and interaction energies of membrane rafts calculated from lipid splay and tilt. AB - Membrane domains known as rafts are rich in cholesterol and sphingolipids, and are thought to be thicker than the surrounding membrane. If so, monolayers should elastically deform so as to avoid exposure of hydrophobic surfaces to water at the raft boundary. We calculated the energy of splay and tilt deformations necessary to avoid such hydrophobic exposure. The derived value of energy per unit length, the line tension gamma, depends on the elastic moduli of the raft and the surrounding membrane; it increases quadratically with the initial difference in thickness between the raft and surround; and it is reduced by differences, either positive or negative, in spontaneous curvature between the two. For zero spontaneous curvature, gamma is approximately 1 pN for a monolayer height mismatch of approximately 0.3 nm, in agreement with experimental measurement. Our model reveals conditions that could prevent rafts from forming, and a mechanism that can cause rafts to remain small. Prevention of raft formation is based on our finding that the calculated line tension is negative if the difference in spontaneous curvature for a raft and the surround is sufficiently large: rafts cannot form if gamma < 0 unless molecular interactions (ignored in the model) are strong enough to make the total line tension positive. Control of size is based on our finding that the height profile from raft to surround does not decrease monotonically, but rather exhibits a damped, oscillatory behavior. As an important consequence, the calculated energy of interaction between rafts also oscillates as it decreases with distance of separation, creating energy barriers between closely apposed rafts. The height of the primary barrier is a complex function of the spontaneous curvatures of the raft and the surround. This barrier can kinetically stabilize the rafts against merger. Our physical theory thus quantifies conditions that allow rafts to form, and further, defines the parameters that control raft merger. PMID- 15542551 TI - Effect of graded hydration on the dynamics of an ion channel peptide: a fluorescence approach. AB - Water plays an important role in determining the folding, structure, dynamics, and, in turn, the function of proteins. We have utilized a combination of fluorescence approaches such as the wavelength-selective fluorescence approach to monitor the effect of varying degrees of hydration on the organization and dynamics of the functionally important tryptophan residues of gramicidin in reverse micelles formed by sodium bis(2-ethylhexyl) sulfosuccinate. Our results show that tryptophans in gramicidin, present in the single-stranded beta6.3 conformation, experience slow solvent relaxation giving rise to red-edge excitation shift (REES). In addition, changes in fluorescence polarization with increasing excitation or emission wavelength reinforce that the gramicidin tryptophans are localized in motionally restricted regions of the reverse micelle. Interestingly, the extent of REES is found to be independent of the [water]/[surfactant] molar ratio (w(o)). We attribute this to heterogeneity in gramicidin tryptophan localization. Fluorescence intensity and mean fluorescence lifetime of the gramicidin tryptophans show significant reductions with increasing w(o) indicating sensitivity to increased polarity. Since the dynamics of hydration is related to folding, structure, and eventually function of proteins, we conclude that REES could prove to be a potentially sensitive tool to explore the dynamics of proteins under conditions of changing hydration. PMID- 15542552 TI - Understanding the shape of sickled red cells. AB - To understand the physical basis of the wide variety of shapes of deoxygenated red cells from patients with sickle cell anemia, we have measured the formation rate and volume distribution of the birefringent domains of hemoglobin S fibers. We find that the domain formation rate depends on the approximately 80th power of the protein concentration, compared to approximately 40th power for the concentration dependence of the reciprocal of the delay time that precedes fiber formation. These remarkably high concentration dependences, as well as the exponential distribution of domain volumes, can be explained by the previously proposed double nucleation model in which homogeneous nucleation of a single fiber triggers the formation of an entire domain via heterogeneous nucleation and growth. The enormous sensitivity of the domain formation rate to intracellular hemoglobin S concentration explains the variable cell morphology and why rapid polymerization results in cells that do not appear sickled at all. PMID- 15542553 TI - Unfolding studies on soybean agglutinin and concanavalin a tetramers: a comparative account. AB - The unfolding pathway of two very similar tetrameric legume lectins soybean agglutinin (SBA) and Concanavalin A (ConA) were determined using GdnCl-induced denaturation. Both proteins displayed a reversible two-state unfolding mechanism. The analysis of isothermal denaturation data provided values for conformational stability of the two proteins. It was found that the DeltaG of unfolding of SBA was much higher than ConA at all the temperatures at which the experiments were done. ConA had a T(g) 18 degrees C less than SBA. The higher conformational stability of SBA in comparison to ConA is largely due to substantial differences in their degrees of subunit interactions. Ionic interactions at the interface of the two proteins especially at the noncanonical interface seem to play a significant role in the observed stability differences between these two proteins. Furthermore, SBA is a glycoprotein with a GlcNac2Man9 chain attached to Asn-75 of each subunit. The sugar chain in SBA lies at the noncanonical interface of the protein, and it is found to interact with the amino acid residues in the adjacent noncanonical interface. These interactions further stabilize SBA with respect to ConA, which is not glycosylated. PMID- 15542554 TI - Asymmetric electron transfer in cyanobacterial Photosystem I: charge separation and secondary electron transfer dynamics of mutations near the primary electron acceptor A0. AB - Point mutations were introduced near the primary electron acceptor sites assigned to A0 in both the PsaA and PsaB branches of Photosystem I in the cyanobacterium Synechocystis sp. PCC 6803. The residues Met688PsaA and Met668PsaB, which provide the axial ligands to the Mg2+ of the eC-A3 and eC-B3 chlorophylls, were changed to leucine and asparagine (chlorophyll notation follows Jordan et al., 2001). The removal of the ligand is expected to alter the midpoint potential of the A0/A0- redox pair and result in a change in the intrinsic charge separation rate and secondary electron transfer kinetics from A0- to A1. The dynamics of primary charge separation and secondary electron transfer were studied at 690 nm and 390 nm in these mutants by ultrafast optical pump-probe spectroscopy. The data reveal that mutations in the PsaB branch do not alter electron transfer dynamics, whereas mutations in the PsaA branch have a distinct effect on electron transfer, slowing down both the primary charge separation and the secondary electron transfer step (the latter by a factor of 3-10). These results suggest that electron transfer in cyanobacterial Photosystem I is asymmetric and occurs primarily along the PsaA branch of cofactors. PMID- 15542555 TI - Normal mode-based fitting of atomic structure into electron density maps: application to sarcoplasmic reticulum Ca-ATPase. AB - A method for the flexible docking of high-resolution atomic structures into lower resolution densities derived from electron microscopy is presented. The atomic structure is deformed by an iterative process using combinations of normal modes to obtain the best fit of the electron microscopical density. The quality of the computed structures has been evaluated by several techniques borrowed from crystallography. Two atomic structures of the SERCA1 Ca-ATPase corresponding to different conformations were used as a starting point to fit the electron density corresponding to a different conformation. The fitted models have been compared to published models obtained by rigid domain docking, and their relation to the known crystallographic structures are explored by normal mode analysis. We find that only a few number of modes contribute significantly to the transition. The associated motions involve almost exclusively rotation and translation of the cytoplasmic domains as well as displacement of cytoplasmic loops. We suggest that the movements of the cytoplasmic domains are driven by the conformational change that occurs between nonphosphorylated and phosphorylated intermediate, the latter being mimicked by the presence of vanadate at the phosphorylation site in the electron microscopy structure. PMID- 15542556 TI - An analysis of core deformations in protein superfamilies. AB - An analysis is presented on how structural cores modify their shape across homologous proteins, and whether or not a relationship exists between these structural changes and the vibrational normal modes that proteins experience as a result of the topological constraints imposed by the fold. A set of 35 representative, well-populated protein families is studied. The evolutionary directions of deformation are obtained by using multiple structural alignments to superimpose the structures and extract a conserved core, together with principal components analysis to extract the main deformation modes from the three dimensional superimposition. In parallel, a low-resolution normal mode analysis technique is employed to study the properties of the mechanical core plasticity of these same families. We show that the evolutionary deformations span a low dimensional space of 4-5 dimensions on average. A statistically significant correspondence exists between these principal deformations and the approximately 20 slowest vibrational modes accessible to a particular topology. We conclude that, to a significant extent, the structural response of a protein topology to sequence changes takes place by means of collective deformations along combinations of a small number of low-frequency modes. The findings have implications in structure prediction by homology modeling. PMID- 15542557 TI - Effect of membrane characteristics on phase separation and domain formation in cholesterol-lipid mixtures. AB - We examine, using an analytical mean-field model, the distribution of cholesterol in a lipid bilayer. The model accounts for the perturbation of lipid packing induced by the embedded cholesterol, in a manner similar to that of transmembrane proteins. We find that the membrane-induced interactions between embedded cholesterol molecules vary as a function of the cholesterol content. Thus, the effective lipid-cholesterol interaction is concentration-dependent. Moreover, it transitions from repulsive to attractive to repulsive as the cholesterol content increases. As the concentration of cholesterol in the bilayer exceeds a critical value, phase separation occurs. The coexistence between cholesterol-rich and cholesterol-poor domains is universal for any bilayer parameters, although the composition of the cholesterol-rich phase varies as a function of the lipid properties. Although we do not assume specific cholesterol-lipid interactions or the formation of a lipid-cholesterol cluster, we find that the composition of the cholesterol-rich domains is constant, independent of the cholesterol content in the bilayer. PMID- 15542558 TI - The dynamic stress responses to area change in planar lipid bilayer membranes. AB - The viscoelastic properties of planar phospholipid (dimyristoylphosphatidylcholine) bilayer membranes at 308 K are studied, many of them for the first time, using the nonequilibrium molecular dynamics simulation (NEMD) method for membrane area change. First, we present a unified formulation of the intrinsic three-dimensional (3D) and apparent in-plane viscoelastic moduli associated with area change based on the constitutive relations for a uniaxial system. The NEMD simulations of oscillatory area change process are then used to obtain the frequency-domain moduli. In the 4-250 GHz range, the intrinsic 3D elastic moduli of 20-27 kbar and viscous moduli of 0.2-9 kbar are found with anisotropy and monotonic frequency dispersion. In contrast, the apparent in-plane elastic moduli (1-9 kbar) are much smaller than, and the viscous moduli (2-6 kbar) comparable to, their 3D counterparts, due to the interplay between the lateral and normal relaxations. The time-domain relaxation functions, separately obtained by applying stepwise strains, can be fit by 4-6 exponential decay modes spanning subpicosecond to nanosecond timescale and are consistent with the frequency-domain results. From NEMD with varying strain amplitude, the linear constitutive model is shown to be valid up to 6 and 20% area change for the intrinsic 3D elastic and viscous responses, respectively, and up to 20% area change for the apparent in-plane viscoelasticity. Inclusion of a gramicidin A dimer (approximately 1 mol %) yields similar response properties with possibly smaller (<10%) viscous moduli. Our results agree well with available data from ultrasonic experiments, and demonstrate that the third dimension (thickness) of the planar lipid bilayer is integral to the in-plane viscoelasticity. PMID- 15542559 TI - Protein structural perturbation and aggregation on homogeneous surfaces. AB - We have demonstrated that globular proteins, such as hen egg lysozyme in phosphate buffered saline at room temperature, lose native structural stability and activity when adsorbed onto well-defined homogeneous solid surfaces. This structural loss is evident by alpha-helix to turns/random during the first 30 min and followed by a slow alpha-helix to beta-sheet transition. Increase in intramolecular and intermolecular beta-sheet content suggests conformational rearrangement and aggregation between different protein molecules, respectively. Amide I band attenuated total reflection/Fourier transformed infrared (ATR/FTIR) spectroscopy was used to quantify the secondary structure content of lysozyme adsorbed on six different self-assembled alkanethiol monolayer surfaces with CH3, -OPh, -CF3, -CN, -OCH3, and -OH exposed functional end groups. Activity measurements of adsorbed lysozyme were in good agreement with the structural perturbations. Both surface chemistry (type of functional groups, wettability) and adsorbate concentration (i.e., lateral interactions) are responsible for the observed structural changes during adsorption. A kinetic model is proposed to describe secondary structural changes that occur in two dynamic phases. The results presented in this article demonstrate the utility of the ATR/FTIR spectroscopic technique for in situ characterization of protein secondary structures during adsorption on flat surfaces. PMID- 15542560 TI - Penetratin-membrane association: W48/R52/W56 shield the peptide from the aqueous phase. AB - Using molecular dynamics simulations, we studied the mode of association of the cell-penetrating peptide penetratin with both a neutral and a charged bilayer. The results show that the initial peptide-lipid association is a fast process driven by electrostatic interactions. The homogeneous distribution of positively charged residues along the axis of the helical peptide, and especially residues K46, R53, and K57, contribute to the association of the peptide with lipids. The bilayer enhances the stability of the penetratin helix. Oriented parallel to the lipid-water interface, the subsequent insertion of the peptide through the bilayer headgroups is significantly slower. The presence of negatively charged lipids considerably enhances peptide binding. Lateral side-chain motion creates an opening for the helix into the hydrophobic core of the membrane. The peptide aromatic residues form a pi-stacking cluster through W48/R52/W56 and F49/R53, protecting the peptide from the water phase. Interaction with the penetratin peptide has only limited effect on the overall membrane structure, as it affects mainly the conformation of the lipids which interact directly with the peptide. Charge matching locally increases the concentration of negatively charged lipids, lateral lipid diffusion locally decreases. Lipid disorder increases, through decreased order parameters of the lipids interacting with the penetratin side chains. Penetratin molecules at the membrane surface do not seem to aggregate. PMID- 15542561 TI - Membrane-protein interactions in mechanosensitive channels. AB - In this article, we examine the mechanical role of the lipid bilayer in ion channel conformation and function with specific reference to the case of the mechanosensitive channel of large conductance (MscL). In a recent article we argued that mechanotransduction very naturally arises from lipid-protein interactions by invoking a simple analytic model of the MscL channel and the surrounding lipid bilayer. In this article, we focus on improving and expanding this analytic framework for studying lipid-protein interactions with special attention to MscL. Our goal is to generate simple scaling relations which can be used to provide qualitative understanding of the role of membrane mechanics in protein function and to quantitatively interpret experimental results. For the MscL channel, we find that the free energies induced by lipid-protein interaction are of the same order as the measured free energy differences between conductance states. We therefore conclude that the mechanics of the bilayer plays an essential role in determining the conformation and function of the channel. Finally, we compare the predictions of our model to experimental results from the recent investigations of the MscL channel by a variety of investigators and suggest a suite of new experiments. PMID- 15542562 TI - Insights into saquinavir resistance in the G48V HIV-1 protease: quantum calculations and molecular dynamic simulations. AB - The spread of acquired immune deficiency syndrome has increasingly become a great concern owing largely to the failure of chemotherapies. The G48V is considered the key signature residue mutation of HIV-1 protease developing with saquinavir therapy. Molecular dynamics simulations of the wild-type and the G48V HIV-1 protease complexed with saquinavir were carried out to explore structure and interactions of the drug resistance. The molecular dynamics results combined with the quantum-based and molecular mechanics Poisson-Boltzmann surface area calculations indicated a monoprotonation took place on D25, one of the triad active site residues. The inhibitor binding of the triad residues and its interaction energy in the mutant were similar to those in the wild-type. The overall structure of both complexes is almost identical. However, the steric conflict of the substituted valine results in the conformational change of the P2 subsite and the disruption of hydrogen bonding between the -NH of the P2 subsite and the backbone -CO of the mutated residue. The magnitude of interaction energy changes was comparable to the experimental K(i) data. The designing for a new drug should consider a reduction of steric repulsion on P2 to enhance the activity toward this mutant strain. PMID- 15542563 TI - Exonuclease I hydrolyzes DNA with a distribution of rates. AB - We report heterogeneity in the time necessary for Exonuclease I to hydrolyze identical DNA fragments. A real-time fluorescence method measured the time required by molecules of Exonuclease I to hydrolyze single-stranded DNA that was synthesized to have two fluorescently labeled nucleotides. One fluorescently labeled nucleotide was located near the 3' end of the DNA and the other near the 5' end. Heterogeneity in the hydrolysis rate of the exonuclease population was inferred from the distribution of times necessary to cleave these DNA fragments. In particular, we found simple first-order kinetics, using a single hydrolysis rate, did not result in a good fit to the data. Better fits to the data were obtained if one assumed a distribution of hydrolysis rates for the exonuclease population. Under our experimental conditions, this broad distribution of rates was centered near 100 nt/s. PMID- 15542564 TI - Dynamic properties of an oriented lipid/DNA complex studied by neutron scattering. AB - The formation of lipid-DNA (CL-DNA) complexes called lipoplexes, proposed as DNA vectors in gene therapy, is obtained by adding DNA to a solution containing liposomes composed of cationic and neutral lipids. The structural and dynamic properties of such lipoplexes are determined by a coupling between the electrostatic interactions and the elastic parameters of the lipid mixture. An attempt to achieve a better understanding of the structure-dynamics relationship is reported herein. In particular, an elastic neutron scattering investigation of DOTAP-DOPC (dioleoyl trimethylammonium propane-dioleoyl phosphatidylcoline) complexed with DNA is described. Proton dynamics in this oriented CL-DNA lipoplex is found to be strongly dependent upon DNA concentration. Our results show that a substantial modification of the membrane dynamics is accompanied by the balancing of the total net charge inside the complex, together with the consequent displacement of interlayer water molecules. PMID- 15542565 TI - Desmin filaments influence myofilament spacing and lateral compliance of slow skeletal muscle fibers. AB - Intermediate filaments composed of desmin interlink Z-disks and sarcolemma in skeletal muscle. Depletion of desmin results in lower active stress of smooth, cardiac, and skeletal muscles. Structural functions of intermediate filaments in fast (psoas) and slow (soleus) skeletal muscle were examined using x-ray diffraction on permeabilized muscle from desmin-deficient mice (Des-/-) and controls (Des+/+). To examine lateral compliance of sarcomeres and cells, filament distances and fiber width were measured during osmotic compression with dextran. Equatorial spacing (x-ray diffraction) of contractile filaments was wider in soleus Des-/- muscle compared to Des+/+, showing that desmin is important for maintaining lattice structure. Osmotic lattice compression was similar in Des-/- and Des+/+. In width measurements of single fibers and bundles, Des-/- soleus were more compressed by dextran compared to Des+/+, showing that intermediate filaments contribute to whole-cell compliance. For psoas fibers, both filament distance and cell compliance were similar in Des-/- and Des+/+. We conclude that desmin is important for stabilizing sarcomeres and maintaining cell compliance in slow skeletal muscle. Wider filament spacing in Des-/- soleus cannot, however, explain the lower active stress, but might influence resistance to stretch, possibly minimizing stretch-induced cell injury. PMID- 15542566 TI - Morphometric changes after thermal and methacholine bronchoprovocations. AB - To determine whether there are distinctions in the location and pattern of response between different bronchoprovocations, we performed high-resolution computer-assisted tomography in 10 asthmatic subjects before and after isocapnic hyperventilation of frigid air (HV) and methacholine (Meth). The luminal areas of the trachea, main stem, lobar, and segmental bronchi were computed before and after each provocation and blindly compared. Both stimuli reduced the 1-s forced expiratory volume similarly (percent change in 1-s forced expiratory volume HV = 28.1 +/- 5.5%, Meth = 25.8 +/- 5.2%; P = 0.69) but did so in different fashions. Each provocation was associated with the development of both bronchial narrowing and dilation; however, more airways constricted with HV (67.7%) than with Meth (47.0%; P < 0.001). Furthermore, there was little concordance between either the magnitude or direction of change between stimuli in any region of the lung (r = 0.25). In general, the frequency of narrowing increased with branching. Constriction became more prominent in the lobar regions and increased further in the segmental branches, but a wide range of intensity existed. These data demonstrate that provocational stimuli evoke complex morphometric changes within the tracheobronchial tree and that different agonists produce different patterns. Thermal stimuli chiefly influence the segmental level, whereas the response to Meth develops more distally. Even within this distribution, the same airway does not respond in an identical fashion to different stimuli, so there does not appear to be a uniform trigger zone. PMID- 15542567 TI - Spike-triggered average torque and muscle fiber conduction velocity of low threshold motor units following submaximal endurance contractions. AB - The motor unit twitch torque is modified by sustained contraction, but the association to changes in muscle fiber electrophysiological properties is not fully known. Thus twitch torque, muscle fiber conduction velocity, and action potential properties of single motor units were assessed in 11 subjects following an isometric submaximal contraction of the tibialis anterior muscle until endurance. The volunteers activated a target motor unit at the minimum discharge rate in eight 3-min-long contractions, three before and five after an isometric contraction at 40% of the maximal torque, sustained until endurance. Multichannel surface electromyogram signals and joint torque were averaged with the target motor unit potential as trigger. Discharge rate (mean +/- SE, 6.6 +/- 0.2 pulses/s) and interpulse interval variability (33.3 +/- 7.0%) were not different in the eight contractions. Peak twitch torque and recruitment threshold increased significantly (93 +/- 29 and 12 +/- 5%, P <0.05) in the contraction immediately after the endurance task with respect to the preendurance values (0.94 +/- 0.26 mN.m and 3.7 +/- 0.5% of the maximal torque), whereas time to peak of the twitch torque did not change (74.4 +/- 10.1 ms). Muscle fiber conduction velocity decreased and action potential duration increased in the contraction after the endurance (6.3 +/- 1.8 and 9.8 +/- 1.8%, respectively, P <0.05; preendurance values, 3.9 +/- 0.2 m/s and 11.1 +/- 0.8 ms), whereas the surface potential peak to-peak amplitude did not change (27.1 +/- 3.1 microV). There was no significant correlation between the relative changes in muscle fiber conduction velocity or surface potential duration and in peak twitch torque (R2= 0.04 and 0.10, respectively). In conclusion, modifications in peak twitch torque of low threshold motor units with sustained contraction are mainly determined by mechanisms not related to changes in action potential shape and in its propagation velocity. PMID- 15542568 TI - Conduction velocity of low-threshold motor units during ischemic contractions performed with surface EMG feedback. AB - The aim of this study was to analyze the effect of ischemia on low-threshold motor unit conduction velocity. Nine subjects were trained to isolate the activity of a single motor unit (target motor unit) in the abductor pollicis brevis muscle with feedback on surface EMG signals recorded with a 16-electrode linear array. After training, the subjects activated the target motor unit at approximately 8 pulses per second (pps) for five 3-min-long contractions. During the third and fourth contractions, a cuff inflated at 180 mmHg around the forearm induced ischemia of the hand. The exerted force (mean +/- SE, 4.6 +/- 2.1% of the maximal voluntary contraction force), discharge rate (8.6 +/- 0.4 pps), interpulse interval variability (34.8 +/- 2.5%), and peak-to-peak amplitude of the target motor unit action potentials (176.6 +/- 18.2 microV) were not different among the five contractions. Conduction velocity, mean power spectral frequency, and action potential duration were the same in the beginning of the five contractions (2.8 +/- 0.2 m/s, 195.2 +/- 10.5 Hz, and 5.4 +/- 0.3 ms, respectively) and changed over the 3 min of sustained activation only during the fourth contraction. Conduction velocity and mean power spectral frequency decreased (10.05 +/- 1.8% and 8.50 +/- 2.18% during the 3 min, respectively) and action potential duration increased (8.2 +/- 4.6% in the 3 min) during the fourth contraction. In conclusion, 1) subjects were able to isolate the activity of a single motor unit with surface EMG visual feedback in ischemic conditions maintained for 16 min, and 2) the activation-induced decrease in single motor unit conduction velocity was significantly larger with ischemia than with normal circulation, probably due to the alteration of mechanisms of ion exchange across the fiber membrane. PMID- 15542569 TI - Dexamethasone treatment in the newborn rat: fatty acid profiling of lung, brain, and serum lipids. AB - Dexamethasone is used as treatment for a variety of neonatal syndromes, including respiratory distress. The present study utilized the power of comprehensive lipid profiling to characterize changes in lipid metabolism in the neonatal lung and brain associated with dexamethasone treatment and also determined the interaction of dexamethasone with hypoxia. A 4-day tapering-dose regimen of dexamethasone was administered at 0800 on postnatal days 3 (0.5 mg/kg), 4 (0.25 mg/kg), 5 (0.125 mg/kg), and 6 (0.05 mg/kg). A subgroup of rats was exposed to hypoxia from birth to 7 days of age. Dexamethasone treatment elicited numerous specific changes in the lipid profile of the normoxic lung, such as increased concentrations of saturated fatty acids in the phosphatidylcholine and cholesterol ester classes. These increases were more profound in the lungs of hypoxic pups. Additional increases in cardiolipin concentrations were also measured in lungs of hypoxic pups treated with dexamethasone. We measured widespread increases in serum lipids after dexamethasone treatment, but the effects were not equivalent between normoxic and hypoxic pups. Dexamethasone treatment in hypoxic pups increased 20:4n6 and 22:6n3 concentrations in the free fatty acid class of the brain. Our results suggest that dexamethasone treatment in neonates elicits specific changes in lung lipid metabolism associated with surfactant production, independent of changes in serum lipids. These findings illustrate the benefits of dexamethasone on lung function but also raise the potential for negative effects due to hyperlipidemia and subtle changes in brain lipid metabolism. PMID- 15542570 TI - IL-6-induced skeletal muscle atrophy. AB - Chronic, low-level elevation of circulating interleukin (IL)-6 is observed in disease states as well as in many outwardly healthy elderly individuals. Increased plasma IL-6 is also observed after intense, prolonged exercise. In the context of skeletal muscle, IL-6 has variously been reported to regulate carbohydrate and lipid metabolism, increase satellite cell proliferation, or cause muscle wasting. In the present study, we used a rodent local infusion model to deliver modest levels of IL-6, comparable to that present after exercise or with chronic low-level inflammation in the elderly, directly into a single target muscle in vivo. The aim of this study was to examine the direct effects of IL-6 on skeletal muscle in the absence of systemic changes in this cytokine. Data included cellular and molecular markers of cytokine and growth factor signaling (phosphorylation and mRNA content) as well as measurements to detect muscle atrophy. IL-6 infusion resulted in muscle atrophy characterized by a preferential loss of myofibrillar protein (-17%). IL-6 induced a decrease in the phosphorylation of ribosomal S6 kinase (-60%) and STAT5 (-33%), whereas that of STAT3 was increased approximately twofold. The changes seen in the IL-6-infused muscles suggest alterations in the balance of growth factor-related signaling in favor of a more catabolic profile. This suggests that downregulation of growth factor-mediated intracellular signaling may be a mechanism contributing to the development of muscle atrophy induced by elevated IL-6. PMID- 15542571 TI - Inter- and intrastrain variation in mouse critical running speed. AB - With the generation of mouse models of human cardiovascular or neuromuscular disorders, the development of noninvasive methods to evaluate the physiological responses to exercise presents an important challenge. The possibility for determining critical speed (CS) in the mouse model was examined according to strain (CD1, C57BL/6J, FVB/N) and sex. Sixty mice performed four exhaustive runs on a treadmill to determine their CS. Twenty-one performed an incremental test to determine the velocity at the lactate threshold. CS was significantly different between the strains (P < 0.0001) but not between sexes. Two measures of heritability showed that CS was partially heritable. CS was not significantly different from lactate threshold velocity. We conclude that CS, which reflects the aerobic capacity, can be determined in mice, as in humans and horses. Considering the intrastrain variability, CS could represent a valuable means for designing an optimal and individualized physical training in mice. PMID- 15542572 TI - Sympathetic outflow enhances the stretch reflex response in the relaxed soleus muscle in humans. AB - Animal experiments suggest that an increase in sympathetic outflow can depress muscle spindle sensitivity and thus modulate the stretch reflex response. The results are, however, controversial, and human studies have failed to demonstrate a direct influence of the sympathetic nervous system on the sensitivity of muscle spindles. We studied the effect of increased sympathetic outflow on the short latency stretch reflex in the soleus muscle evoked by tapping the Achilles tendon. Nine subjects performed three maneuvers causing a sustained activation of sympathetic outflow to the leg: 3 min of static handgrip exercise at 30% of maximal voluntary contraction, followed by 3 min of posthandgrip ischemia, and finally during a 3-min mental arithmetic task. Electromyography was measured from the soleus muscle with bipolar surface electrodes during the Achilles tendon tapping, and beat-to-beat changes in heart rate and mean arterial blood pressure were monitored continuously. Mean arterial pressure was significantly elevated during all three maneuvers, whereas heart rate was significantly elevated during static handgrip exercise and mental arithmetic but not during posthandgrip ischemia. The peak-to-peak amplitude of the short-latency stretch reflex was significantly increased during mental arithmetic (P < 0.05), static handgrip exercise (P < 0.001), and posthandgrip ischemia (P < 0.005). When expressed in percent change from rest, the mean peak-to-peak amplitude increased by 111 (SD 100)% during mental arithmetic, by 160 (SD 103)% during static handgrip exercise, and by 90 (SD 67)% during posthandgrip ischemia. The study clearly indicates a facilitation of the short-latency stretch reflex during increased sympathetic outflow. We note that the enhanced stretch reflex responses observed in relaxed muscles in the absence of skeletomotor activity support the idea that the sympathetic nervous system can exert a direct influence on the human muscle spindles. PMID- 15542573 TI - Elevated body temperature enhances the laryngeal chemoreflex in decerebrate piglets. AB - Hyperthermia and reflex apnea may both contribute to sudden infant death syndrome (SIDS). Therefore, we investigated the effect of increased body temperature on the inhibition of breathing produced by water injected into the larynx, which elicits the laryngeal chemoreflex (LCR). We studied decerebrated, vagotomized, neonatal piglets aged 3-15 days. Blood pressure, end-tidal CO(2), body temperature, and phrenic nerve activity were recorded. To elicit the LCR, we infused 0.1 ml of distilled water through a polyethylene tube passed through the nose and positioned just rostral to the larynx. Three to five LCR trials were performed with the piglet at normal body temperature. The animal's core body temperature was raised by approximately 2.5 degrees C, and three to five LCR trials were performed before the animal was cooled, and three to five LCR trials were repeated. The respiratory inhibition associated with the LCR was substantially prolonged when body temperature was elevated. Thus elevated body temperature may contribute to the pathogenesis of SIDS by increasing the inhibitory effects of the LCR. PMID- 15542574 TI - Two temporal components within the human pulmonary vascular response to approximately 2 h of isocapnic hypoxia. AB - The time course of the pulmonary vascular response to hypoxia in humans has not been fully defined. In this investigation, study A was designed to assess the form of the increase in pulmonary vascular tone at the onset of hypoxia and to determine whether a steady plateau ensues over the following approximately 20 min. Twelve volunteers were exposed twice to 5 min of isocapnic euoxia (end-tidal Po(2) = 100 Torr), 25 min of isocapnic hypoxia (end-tidal Po(2) = 50 Torr), and finally 5 min of isocapnic euoxia. Study B was designed to look for the onset of a slower pulmonary vascular response, and, if possible, to determine a latency for this process. Seven volunteers were exposed to 5 min of isocapnic euoxia, 105 min of isocapnic hypoxia, and finally 10 min of isocapnic euoxia. For both studies, control protocols consisting of isocapnic euoxia were undertaken. Doppler echocardiography was used to measure cardiac output and the maximum tricuspid pressure gradient during systole, and estimates of pulmonary vascular resistance were calculated. For study A, the initial response was well described by a monoexponential process with a time constant of 2.4 +/- 0.7 min (mean +/- SE). After this, there was a plateau phase lasting at least 20 min. In study B, a second slower phase was identified, with vascular tone beginning to rise again after a latency of 43 +/- 5 min. These findings demonstrate the presence of two distinct phases of hypoxic pulmonary vasoconstriction, which may result from two distinct underlying processes. PMID- 15542575 TI - A fractal analysis of the radial distribution of bronchial capillaries around large airways. AB - We analyzed published measurements of the bronchial circulation and airway wall (Anderson JC, Bernard SL, Luchtel DL, Babb AL, and Hlastala MP. Respir Physiol Neurobiol 132: 329-339, 2002) and determined that the radial distribution of bronchial capillary cross-sectional area was fractal. We limited our analysis to bronchial capillaries, diameter < or =10 mum, that resided between the epithelial basement membrane and adventitia-alveolar boundary, the airway wall tissue. Thirteen different radial distributions of capillary-to-tissue area were constructed simply by changing the number of annuli (i.e., the annular size) used to form each distribution. For the 13 distributions created, these annuli ranged in size from to of the size of the airway wall area. Radial distributions were excluded from the fractal analysis if the sectioning procedure resulted in an annulus with a radial thickness less than the diameter of a capillary. To determine the fractal dimension for a given airway, the coefficient of variation (CV) for each distribution was calculated, and ln(CV) was plotted against the logarithm of the relative piece area. For airways with diameter >2.4 mm, this relationship was linear, which indicated the radial distribution of bronchial capillary cross-sectional area was fractal with an average fractal dimension of 1.27. The radial distribution of bronchial capillary cross-sectional area was not fractal around airways with diameter <1.5 mm. We speculated on how the fractal nature of this circulation impacts the distribution of bronchial blood flow and the efficiency of mass transport during health and disease. A fractal analysis can be used as a tool to quantify and summarize investigations of the bronchial circulation. PMID- 15542576 TI - Interaction between the canine diaphragm and intercostal muscles in lung expansion. AB - Changes in intrathoracic pressure produced by the various inspiratory intercostals are essentially additive, but the interaction between these muscles and the diaphragm remains uncertain. In the present study, this interaction was assessed by measuring the changes in airway opening (DeltaPao) or transpulmonary pressure (DeltaPtp) in vagotomized, phrenicotomized dogs during spontaneous inspiration (isolated intercostal contraction), during isolated rectangular or ramp stimulation of the peripheral ends of the transected C(5) phrenic nerve roots (isolated diaphragm contraction), and during spontaneous inspiration with superimposed phrenic nerve stimulation (combined diaphragm-intercostal contraction). With the endotracheal tube occluded at functional residual capacity, DeltaPao during combined diaphragm-intercostal contraction was nearly equal to the sum of the DeltaPao produced by the two muscle groups contracting individually. However, when the endotracheal tube was kept open, DeltaPtp during combined contraction was 123% of the sum of the individual DeltaPtp (P < 0.001). The increase in lung volume during combined contraction was also 109% of the sum of the individual volume increases (P < 0.02). Abdominal pressure during combined contraction was invariably lower than during isolated diaphragm contraction. It is concluded, therefore, that the canine diaphragm and intercostal muscles act synergistically during lung expansion and that this synergism is primarily due to the fact that the intercostal muscles reduce shortening of the diaphragm. When the lung is maintained at functional residual capacity, however, the synergism is obscured because the greater stiffness of the rib cage during diaphragm contraction enhances the DeltaPao produced by the isolated diaphragm and reduces the DeltaPao produced by the intercostal muscles. PMID- 15542577 TI - Previous exercise attenuates muscle sympathetic activity and increases blood flow during acute euglycemic hyperinsulinemia. AB - Insulin infusion causes muscle vasodilation, despite the increase in sympathetic nerve activity. In contrast, a single bout of exercise decreases sympathetic activity and increases muscle blood flow during the postexercise period. We tested the hypothesis that muscle sympathetic activity would be lower and muscle vasodilation would be higher during hyperinsulinemia performed after a single bout of dynamic exercise. Twenty-one healthy young men randomly underwent two hyperinsulinemic euglycemic clamps performed after 45 min of seated rest (control) or bicycle exercise (50% of peak oxygen uptake). Muscle sympathetic nerve activity (MSNA, microneurography), forearm blood flow (FBF, plethysmography), blood pressure (BP, oscillometric method), and heart rate (HR, ECG) were measured at baseline (90 min after exercise or seated rest) and during hyperinsulinemic euglycemic clamps. Baseline glucose and insulin concentrations were similar in the exercise and control sessions. Insulin sensitivity was unchanged by previous exercise. During the clamp, insulin levels increased similarly in both sessions. As expected, insulin infusion increased MSNA, FBF, BP, and HR in both sessions (23 +/- 1 vs. 36 +/- 2 bursts/min, 1.8 +/- 0.1 vs. 2.2 +/- 0.2 ml.min(-1).100 ml(-1), 89 +/- 2 vs. 92 +/- 2 mmHg, and 58 +/- 1 vs. 62 +/- 1 beats/min, respectively, P < 0.05). BP and HR were similar between sessions. However, MSNA was significantly lower (27 +/- 2 vs. 31 +/- 2 bursts/min), and FBF was significantly higher (2.2 +/- 0.2 vs. 1.8 +/- 0.1 ml.min(-1).100 ml(-1), P < 0.05) in the exercise session compared with the control session. In conclusion, in healthy men, a prolonged bout of dynamic exercise decreases MSNA and increases FBF. These effects persist during acute hyperinsulinemia performed after exercise. PMID- 15542578 TI - Unmasking Evans syndrome: T-cell phenotype and apoptotic response reveal autoimmune lymphoproliferative syndrome (ALPS). AB - Autoimmune lymphoproliferative syndrome (ALPS) is a rare disorder of disrupted lymphocyte homeostasis. Clinical manifestations of ALPS vary but typically include autoimmune cytopenias, organomegaly, lymphadenopathy, and increased risk of malignancies. A similar spectrum of symptoms may be seen in some patients with Evans syndrome (ES), a hematologic disorder defined by autoimmune destruction of at least 2 hematologic cell types. We hypothesized that a subset of patients diagnosed with ES may have ALPS. We screened 12 children with ES by flow cytometric analysis for CD4-/CD8- (double negative) T cells (DNTs) and with the definitive test for ALPS, defective in vitro Fas-mediated apoptosis. Six of the patients had elevated DNTs, suggestive of ALPS and also had defective Fas mediated apoptosis. The other 6 patients displayed normal T-cell apoptosis; 5 of whom had normal DNTs, and 1 had a borderline result. Thus, 7 (58%) of 12 patients with ES had elevated DNTs suggestive of ALPS, with functional confirmation in 6 of 7. This suggests that analysis of DNTs may be a sensitive first-line screening test, serving as a marker of patients who should undergo definitive testing for ALPS. Our data further suggest that a number of patients with ES may have ALPS, a novel finding with important therapeutic implications. PMID- 15542579 TI - The CXC chemokine MIP-2 stimulates neutrophil mobilization from the rat bone marrow in a CD49d-dependent manner. AB - The acute release of neutrophils from the bone marrow is a critical step in their trafficking to sites of inflammation. This process is stimulated by systemically acting inflammatory mediators, such as the CXC chemokines. In this study we have used a novel in situ perfusion system of the rat femoral bone marrow to directly investigate the role of specific adhesion molecules in chemokine-stimulated neutrophil mobilization. We show here that neutrophils mobilized in response to rat macrophage inflammatory protein-2 (MIP-2) shed L-selectin and expressed significantly higher levels of CD11b and CD49d. However, inhibition of L-selectin sheddase activity with KD-IX-73-4 had no effect on the number of neutrophils mobilized in response to rat MIP-2. Blockade of CD18, using a neutralizing monoclonal antibody (mAb), did not inhibit neutrophil mobilization but unexpectedly increased the rate and number of neutrophils released from the bone marrow in response to chemokine, suggesting that CD18 could play a role in neutrophil retention within the bone marrow. Blockade of CD49d using either a selective mAb or a specific antagonist resulted in a dramatic inhibition (> 75%) of the chemokine-stimulated neutrophil mobilization from the bone marrow. These data reveal contrasting roles for CD18 and CD49d in the retention and release of neutrophils from the bone marrow. PMID- 15542580 TI - Increased peripheral platelet destruction and caspase-3-independent programmed cell death of bone marrow megakaryocytes in myelodysplastic patients. AB - To investigate underlying mechanisms of thrombocytopenia in myelodysplastic syndrome (MDS), radiolabeled platelet studies were performed in 30 MDS patients with platelet counts less than 100 x 10(9)/L. Furthermore, plasma thrombopoietin and glycocalicin index (a parameter of platelet or megakaryocyte destruction) were determined. Mean platelet life (MPL), corrected for the degree of thrombocytopenia, was reduced in 15 of 30 patients (4.3 +/- 0.9 days [mean +/- SD] vs 6.0 +/- 1.3, P = .0003). Platelet production rate (PPR) was reduced in 25 of 30 patients (68 +/- 34 x 10(9)/d vs 220 +/- 65, P < .0001). Thrombopoietin levels were not significantly correlated with the PPR. However, the glycocalicin index was significantly higher compared with controls (15 +/- 16 vs 0.7 +/- 0.2, P = .001) and significantly correlated with the PPR (P = .02, r = -0.5), but not with the MPL (P = 1.8). Ultrastructural studies demonstrated necrosis-like programmed cell death (PCD) in mature and immature megakaryocytes (n = 9). Immunohistochemistry of the bone marrow biopsies demonstrated no positive staining of MDS megakaryocytes for activated caspase-3 (n = 24) or cathepsin D (n = 21), while activated caspase-8 was demonstrated in a subgroup of patients (5/21) in less than 10% of megakaryocytes. These results indicate that the main cause of thrombocytopenia in MDS is caspase-3-independent necrosis-like PCD resulting in a decreased PPR in conjunction with an increased glycocalicin index. PMID- 15542581 TI - PKCdelta plays opposite roles in growth mediated by wild-type Kit and an oncogenic Kit mutant. AB - The Kit receptor tyrosine kinase is critical for normal hematopoiesis. Mutation of the aspartic acid residue encoded by codon 816 of human c-kit or codon 814 of the murine gene results in an oncogenic form of Kit. Here we investigate the role of protein kinase Cdelta (PKCdelta) in responses mediated by wild-type murine Kit and the D814Y mutant in a murine mast cell-like line. PKCdelta is activated after wild-type (WT) Kit binds stem cell factor (SCF), is constitutively active in cells expressing the Kit catalytic domain mutant, and coprecipitates with both forms of Kit. Inhibition of PKCdelta had opposite effects on growth mediated by wild-type and mutant Kit. Both rottlerin and a dominant-negative PKCdelta construct inhibited the growth of cells expressing mutant Kit, while SCF-induced growth of cells expressing wild-type Kit was not inhibited. Further, overexpression of PKCdelta inhibited growth of cells expressing wild-type Kit and enhanced growth of cells expressing the Kit mutant. These data demonstrate that PKCdelta contributes to factor-independent growth of cells expressing the D814Y mutant, but negatively regulates SCF-induced growth of cells expressing wild-type Kit. This is the first demonstration that PKCdelta has different functions in cells expressing normal versus oncogenic forms of a receptor. PMID- 15542582 TI - Promoter trapping reveals significant differences in integration site selection between MLV and HIV vectors in primary hematopoietic cells. AB - Recent reports have indicated that human immunodeficiency virus (HIV) and murine leukemia virus (MLV) vectors preferentially integrate into active genes. Here, we used a novel approach based on genetic trapping to rapidly score several thousand integration sites and found that MLV vectors trapped cellular promoters more efficiently than HIV vectors. Remarkably, 1 in 5 MLV integrations trapped an active promoter in different cell lines and primary hematopoietic cells. Such frequency was even higher in growth-stimulated lymphocytes. We show that the different behavior of MLV and HIV vectors was dependent on a different integration pattern within transcribed genes. Whereas MLV-based traps showed a strong bias for promoter-proximal integration leading to efficient reporter expression, HIV-based traps integrated throughout transcriptional units and were limited for expression by the distance from the promoter and the reading frame of the targeted gene. Our results indicate a strong propensity of MLV to establish transcriptional interactions with cellular promoters, a behavior that may have evolved to enhance proviral expression and may increase the insertional mutagenesis risk. Promoter trapping efficiency provides a convenient readout to assess transcriptional interactions between the vector and its flanking genes at the integration site and to compare integration site selection among different cell types and in different growth conditions. PMID- 15542583 TI - Treatment of nasopharyngeal carcinoma with Epstein-Barr virus--specific T lymphocytes. AB - Conventional treatment for nasopharyngeal carcinoma (NPC) frequently fails and is accompanied by severe long-term side effects. Since virtually all undifferentiated NPCs are associated with Epstein-Barr virus (EBV), this tumor is an attractive candidate for cellular immunotherapy targeted against tumor associated viral antigens. We now demonstrate that EBV-specific cytotoxic T-cell (CTL) lines can readily be generated from individuals with NPC, notwithstanding the patients' prior exposure to chemotherapy/radiation. A total of 10 patients diagnosed with advanced NPC were treated with autologous CTLs. All patients tolerated the CTLs, although one developed increased swelling at the site of pre existing disease. At 19 to 27 months after infusion, 4 patients treated in remission from locally advanced disease remain disease free. Of 6 patients with refractory disease prior to treatment, 2 had complete responses, and remain in remission over 11 to 23 months after treatment; 1 had a partial remission that persisted for 12 months; 1 has had stable disease for more than 14 months; and 2 had no response. These results demonstrate that administration of EBV-specific CTLs to patients with advanced NPC is feasible, appears to be safe, and can be associated with significant antitumor activity. PMID- 15542584 TI - Immunoproliferative small intestinal disease (IPSID): a model for mature B-cell neoplasms. AB - Immunoproliferative small intestinal disease (IPSID) was recently added to the growing list of infectious pathogen-associated human lymphomas. Molecular and immunohistochemical studies demonstrated an association with Campylobacter jejuni. IPSID is a variant of the B-cell lymphoma of mucosa-associated lymphoid tissue (MALT), which involves mainly the proximal small intestine resulting in malabsorption, diarrhea, and abdominal pain. Geographically, IPSID is most prevalent in the Middle East and Africa. IPSID lymphomas reveal excessive plasma cell differentiation and produce truncated alpha heavy chain proteins lacking the light chains as well as the first constant domain. The corresponding mRNA lacks the variable heavy chain (V(H)) and the constant heavy chain 1 (C(H)1) sequences and contains deletions as well as insertions of unknown origin. The encoding gene sequence reveals a deletion of V region and parts of C(H)1 domain. Cytogenetic studies demonstrated clonal rearrangements involving predominantly the heavy and light chain genes, including t(9;14) translocation involving the PAX5 gene. Early stage IPSID responds to antibiotics (30%-70% complete remission). Most untreated IPSID patients progress to lymphoplasmacytic and immunoblastic lymphoma invading the intestinal wall and mesenteric lymph nodes, and may metastasize to a distant organ. IPSID lymphoma shares clinical, morphologic, and molecular features with MALT lymphoma, lymphoplasmacytic lymphoma, and plasma cell neoplasms. PMID- 15542585 TI - Rapid and large amount of autocrine IL-3 production is responsible for mast cell survival by IgE in the absence of antigen. AB - Cross-linking FcepsilonRI on mast cells by immunoglobulin E (IgE) and antigen (Ag) initiates cascades leading to antiparasitic or allergic responses. It was recently reported that IgE without antigen, IgE(-Ag), actively promotes mast cell survival. Although we have demonstrated that the immunoreceptor tyrosine-based activation motif within FcRgamma is essential for IgE(-Ag)-induced mast cell survival, the underlying mechanism remains still unclear. Here, we investigated the mechanism of IgE(-Ag)-induced survival using mast cells lacking several downstream molecules. Lyn and Syk were essential, whereas Fyn, Gab2, and the phosphoinositide 3-kinase-Akt pathway were not critical for survival. Failure of survival in FcRgamma-/- bone marrow mast cells (BMMCs) was rescued by coculture with IgE-treated wild-type BMMCs, suggesting that survival is induced not directly through FcepsilonRI signals. We found that the survival is predominantly mediated by high production of interleukin 3 (IL-3), evidenced by severe impairment of survival by anti-IL-3 and in IL-3-/- BMMCs. The up-regulation of Bcl-xL/Bcl-2 by IgE was abrogated in IL-3-/- BMMCs, whereas the expression of histidine decarboxylase was normally induced. These results indicate that IL-3 plays a crucial role for IgE(-Ag)-induced mast cell survival, functioning in an autocrine manner by inducing the Bcl-xL/Bcl-2 via signal transducer and activator of transduction 5. We further suggest that IgE(-Ag)-mediated gene expression in mast cells is regulated at least 2 mechanisms: autocrine IL-3 dependent and independent. PMID- 15542586 TI - Treatment of acquired hemophilia by the Bonn-Malmo Protocol: documentation of an in vivo immunomodulating concept. AB - Acquired hemophilia (AH) is an extremely rare condition in which autoantibodies (inhibitors) against clotting factor VIII induce acute and life-threatening hemorrhagic diathesis because of abnormal blood clotting. The mortality rate of AH is as high as 16%, and current treatment options are associated with adverse side effects. We investigated a therapeutic approach for AH called the modified Bonn-Malmo Protocol (MBMP). The aims of MBMP include suppression of bleeding, permanent elimination of inhibitors, and development of immune tolerance, thereby avoiding long-term reliance on coagulation products. The protocol included immunoadsorption for inhibitor elimination, factor VIII substitution, intravenous immunoglobulin, and immunosuppression. Thirty-five high-titer patients with critical bleeding who underwent MBMP were evaluated. Bleeding was rapidly controlled during 1 or 2 apheresis sessions, and no subsequent bleeding episodes occurred. Inhibitor levels decreased to undetectable levels within a median of 3 days (95% confidence interval [95% CI], 2-4 days), factor substitution was stopped within a median of 12 days (95% CI, 11-17 days), and treatment was completed within a median of 14 days (95% CI, 12-17 days). Long-term follow-up (7 months-7 years) showed an overall response rate of 88% for complete remission (CR). When cancer patients were excluded, the CR rate was 97%. PMID- 15542587 TI - Transient expansion of synaptically connected dendritic spines upon induction of hippocampal long-term potentiation. AB - Dendritic spines are small protrusions from dendritic shafts that contain the postsynaptic sites of glutamatergic synapses in the brain. Spines undergo dramatic activity-dependent structural changes that are particularly prominent during neuronal development. Although changes in spine shape or number have been proposed to contribute to forms of synaptic plasticity that underlie learning and memory, the extent to which spines remain plastic in the adult brain is unclear. We find that induction of long-term potentiation (LTP) of synaptic transmission in acute hippocampal slices of adult mice evokes a reliable, transient expansion in spines that are synaptically activated, as determined with calcium imaging. Similar to LTP, transient spine expansion requires N-methyl-D-aspartate (NMDA) receptor-mediated Ca2+ influx and actin polymerization. Moreover, like the early phase of LTP induced by the stimulation protocol, spine expansion does not require Ca2+ influx through L-type voltage-gated Ca2+ channels nor does it require protein synthesis. Thus, transient spine expansion is a characteristic feature of the initial phases of plasticity at mature synapses and so may contribute to synapse remodeling important for LTP. PMID- 15542588 TI - A dual-genome Symbiosis Chip for coordinate study of signal exchange and development in a prokaryote-host interaction. AB - The soil-dwelling alpha-proteobacterium Sinorhizobium meliloti engages in a symbiosis with legumes: S. meliloti elicits the formation of plant root nodules where it converts dinitrogen to ammonia for use by the plant in exchange for plant photosynthate. To study the coordinate differentiation of S. meliloti and its legume partner during nodule development, we designed a custom Affymetrix GeneChip with the complete S. meliloti genome and approximately 10,000 probe sets for the plant host, Medicago truncatula. Expression profiling of free-living S. meliloti grown with the plant signal molecule luteolin in defined minimal and rich media or of strains altered in the expression of key regulatory proteins (NodD1, NodD3, and RpoN) confirms previous data and identifies previously undescribed regulatory targets. Analyses of root nodules show that this Symbiosis Chip allows the study of gene expression in both partners simultaneously. Our studies detail nearly 5,000 transcriptome changes in symbiosis and document complex transcriptional profiles of S. meliloti in different environments. PMID- 15542589 TI - LIM domains-containing protein 1 (LIMD1), a tumor suppressor encoded at chromosome 3p21.3, binds pRB and represses E2F-driven transcription. AB - LIM domains-containing protein 1 (LIMD1) is encoded at chromosome 3p21.3, a region commonly deleted in many solid malignancies. However, the function of LIMD1 is unknown. Here we show that LIMD1 specifically interacts with retinoblastoma protein (pRB), inhibits E2F-mediated transcription, and suppresses the expression of the majority of genes with E2F1-responsive elements. LIMD1 blocks tumor growth in vitro and in vivo and is down-regulated in the majority of human lung cancer samples tested. Our data indicate that LIMD1 is a tumor suppressor gene, the protein product of which functionally interacts with pRB and the loss of which promotes lung carcinogenesis. PMID- 15542590 TI - The crystal structure of guinea pig 11beta-hydroxysteroid dehydrogenase type 1 provides a model for enzyme-lipid bilayer interactions. AB - The metabolic reduction of 11-keto groups in glucocorticoid steroids such as cortisone leads to the nuclear receptor ligand cortisol. This conversion is an example of pre-receptor regulation and constitutes a novel pharmacological target for the treatment of metabolic disorders such as insulin resistance and possibly other derangements observed in the metabolic syndrome, such as hyperlipidemia, hypertension, and lowered insulin secretion. This reaction is carried out by the NADPH-dependent type 1 11beta-hydroxysteroid dehydrogenase (11beta-HSD1), an enzyme attached through an integral N-terminal transmembrane helix to the lipid bilayer and located with its active site within the lumen of the endoplasmic reticulum. Here we report the crystal structure of recombinant guinea pig 11beta HSD1. This variant was determined in complex with NADP at 2.5 A resolution and crystallized in the presence of detergent and guanidinium hydrochloride. The overall structure of guinea pig 11beta-HSD1 shows a clear relationship to other members of the superfamily of short-chain dehydrogenases/reductases but harbors a unique C-terminal helical segment that fulfills three essential functions and accordingly is involved in subunit interactions, contributes to active site architecture, and is necessary for lipid-membrane interactions. The structure provides a model for enzyme-lipid bilayer interactions and suggests a funneling of lipophilic substrates such as steroid hormones from the hydrophobic membrane environment to the enzyme active site. PMID- 15542591 TI - Residues 10-18 within the C5a receptor N terminus compose a binding domain for chemotaxis inhibitory protein of Staphylococcus aureus. AB - Chemotaxis inhibitory protein of Staphylococcus aureus (CHIPS) is excreted by the majority of S. aureus strains and is a potent inhibitor of C5a- and formylated peptide-mediated chemotaxis of neutrophils and monocytes. Recently, we reported that CHIPS binds to the C5a receptor (C5aR) and the formylated peptide receptor, thereby blocking activation by C5a and formylated peptides, respectively. The anaphylatoxin C5a plays an important role in host immunity and pathological inflammatory processes. For C5a a two-site binding model is proposed in which C5a initially binds the C5aR N terminus, followed by interaction of the C5a C terminal tail with an effector domain on the receptor. We have shown here that CHIPS does not affect activation of the C5aR by a peptide mimic of the C5a C terminus. Moreover, CHIPS was found to bind human embryonic kidney 293 cells expressing only the C5aR N terminus. Deletion and mutation experiments within this C5aR N-terminal expression system revealed that the binding site of CHIPS is contained in a short stretch of 9 amino acids (amino acids 10-18), of which the aspartic acid residues at positions 10, 15, and 18 plus the glycine at position 12 are crucial. Binding studies with C5aR/C3aR and C5aR/IL8RA chimeras confirmed that CHIPS binds only to the C5aR N terminus without involvement of its extracellular loops. CHIPS may provide new strategies to block the C5aR, which may lead to the development of new C5aR antagonists. PMID- 15542592 TI - Structures of APRIL-receptor complexes: like BCMA, TACI employs only a single cysteine-rich domain for high affinity ligand binding. AB - TACI is a member of the tumor necrosis factor receptor superfamily and serves as a key regulator of B cell function. TACI binds two ligands, APRIL and BAFF, with high affinity and contains two cysteine-rich domains (CRDs) in its extracellular region; in contrast, BCMA and BR3, the other known high affinity receptors for APRIL and BAFF, respectively, contain only a single or partial CRD. However, another form of TACI exists wherein the N-terminal CRD is removed by alternative splicing. We find that this shorter form is capable of ligand-induced cell signaling and that the second CRD alone (TACI_d2) contains full affinity for both ligands. Furthermore, we report the solution structure and alanine-scanning mutagenesis of TACI_d2 along with co-crystal structures of APRIL.TACI_d2 and APRIL.BCMA complexes that together reveal the mechanism by which TACI engages high affinity ligand binding through a single CRD, and we highlight sources of ligand-receptor specificity within the APRIL/BAFF system. PMID- 15542593 TI - The dileucine motif at the COOH terminus of human multidrug resistance P glycoprotein is important for folding but not activity. AB - P-glycoprotein (P-gp, ABCB1) actively transports a broad range of cytotoxic compounds out of the cell. The COOH terminus of P-gp contains a dileucine motif (Leu(1260)-Leu(1261)) and a conserved phenylalanine (Phe(1268)). Similar residues in SUR1 (ABCC8) were reported to be important plasma membrane-targeting signals (Sharma, N., Crane, A., Clement, J. P. t., Gonzalez, G., Babenko, A. P., Bryan, J., and Aguilar-Bryan, L. (1999) J. Biol. Chem. 274, 20628-20632). Here, we used alanine-scanning mutagenesis to test whether these residues were essential for trafficking of P-gp to the cell surface. Mutant L1260A expressed a 150-kDa immature protein that did not reach the cell surface and was sensitive to digestion by Endo H(f). By contrast, mutants L1261A, F1268A, and wild-type P-gps expressed the 170-kDa mature proteins at the cell surface. Mutation of Leu(1260) to Gly, Ile, Trp, Lys, or Glu also resulted in the expression of the 150-kDa immature protein. All of the mutants, however, expressed the 170-kDa protein in the presence of the drug substrate/specific chemical chaperone cyclosporin A. Mutant L1260A P-gp exhibited drug-stimulated ATPase activities similar to that of wild-type enzyme after rescue with cyclosporin A. Deletion of the last 22 amino acids (Q(1259)-Q(1280)) also caused misprocessing. The mutant, however, was rescued by expression in the presence of cyclosporin A and conferred resistance to colchicine in transfected cells. These results show that the dileucine motif is not a plasma membrane targeting signal. The COOH terminus is required for proper folding of P-gp but not for activity. PMID- 15542594 TI - Crystal structures of novel vascular endothelial growth factors (VEGF) from snake venoms: insight into selective VEGF binding to kinase insert domain-containing receptor but not to fms-like tyrosine kinase-1. AB - Vascular endothelial growth factor-A (VEGF-A(165)) exerts multiple effects upon binding to the fms-like tyrosine kinase-1 (Flt-1) and the kinase insert domain containing receptor (KDR). We recently identified two novel snake venom VEGFs (vammin and VR-1) having unique properties. These VEGFs, designated VEGF-Fs, are highly specific ligands for the kinase insert domain-containing receptor and exhibit potent biological activity both in vitro and in vivo when compared with VEGF-A(165). Here, we solved the crystal structures of vammin and VR-1 at 1.9 and 2.0 A resolutions, respectively. Both structures are very similar to each other, and these structures exhibit similar but significantly different features from the known structures of other VEGFs. These differences include a conformational difference in receptor-binding loop 3 caused by an amino acid residue insertion and a difference in surface potential on the possible binding surface for domain 3 of the receptor. These structural differences may be related to the highly selective ligand properties of VEGF-F. PMID- 15542595 TI - Succinate complex crystal structures of the alpha-ketoglutarate-dependent dioxygenase AtsK: steric aspects of enzyme self-hydroxylation. AB - The alkylsulfatase AtsK from Pseudomonas putida S-313 is a member of the non-heme iron(II)-alpha-ketoglutarate-dependent dioxygenase superfamily. In the initial step of their catalytic cycle, enzymes belonging to this widespread and versatile family coordinate molecular oxygen to the iron center in the active site. The subsequent decarboxylation of the cosubstrate alpha-ketoglutarate yields carbon dioxide, succinate, and a highly reactive ferryl (IV) species, which is required for substrate oxidation via a complex mechanism involving the transfer of radical species. Non-productive activation of oxygen may lead to harmful side reactions; therefore, such enzymes need an effective built-in protection mechanism. One of the ways of controlling undesired side reactions is the self-hydroxylation of an aromatic side chain, which leads to an irreversibly inactivated species. Here we describe the crystal structure of the alkylsulfatase AtsK in complexes with succinate and with Fe(II)/succinate. In the crystal structure of the AtsK-Fe(II) succinate complex, the side chain of Tyr(168) is co-ordinated to the iron, suggesting that Tyr(168) is the target of enzyme self-hydroxylation. This is the first structural study of an Fe(II)-alpha-ketoglutarate-dependent dioxygenase that presents an aromatic side chain coordinated to the metal center, thus allowing structural insight into this protective mechanism of enzyme self inactivation. PMID- 15542597 TI - Structural changes of D1 C-terminal alpha-carboxylate during S-state cycling in photosynthetic oxygen evolution. AB - Changes in the chemical structure of alpha-carboxylate of the D1 C-terminal Ala 344 during S-state cycling of photosynthetic oxygen-evolving complex were selectively measured using light-induced Fourier transform infrared (FTIR) difference spectroscopy in combination with specific [(13)C]alanine labeling and site-directed mutagenesis in photosystem II core particles from Synechocystis sp. PCC 6803. Several bands for carboxylate symmetric stretching modes in an S(2)/S(1) FTIR difference spectrum were affected by selective (13)C labeling of the alpha-carboxylate of Ala with l-[1-(13)C]alanine, whereas most of the isotopic effects failed to be induced in a site-directed mutant in which Ala-344 was replaced with Gly. Labeling of the alpha-methyl of Ala with l-[3 (13)C]alanine had much smaller effects on the spectrum to induce isotopic bands due to a symmetric CH(3) deformation coupled with the alpha-carboxylate. The isotopic bands for the alpha-carboxylate of Ala-344 showed characteristic changes during S-state cycling. The bands appeared prominently upon the S(1)-to-S(2) transition and to a lesser extent upon the S(2)-to-S(3) transition but reappeared at slightly upshifted frequencies with the opposite sign upon the S(3)-to-S(0) transition. No obvious isotopic band appeared upon the S(0)-to-S(1) transition. These results indicate that the alpha-carboxylate of C-terminal Ala-344 is structurally associated with a manganese ion that becomes oxidized upon the S(1) to-S(2) transition and reduced reversely upon the S(3)-to-S(0) transition but is not associated with manganese ion(s) oxidized during the S(0)-to-S(1) (and S(2) to-S(3)) transition(s). Consistently, l-[1-(13)C]alanine labeling also induced spectral changes in the low frequency (670-350 cm(-1)) S(2)/S(1) FTIR difference spectrum. PMID- 15542596 TI - The SNARE proteins SNAP-25 and SNAP-23 display different affinities for lipid rafts in PC12 cells. Regulation by distinct cysteine-rich domains. AB - SNAP-25 and its ubiquitously expressed homologue, SNAP-23, are SNARE proteins that are essential for regulated exocytosis in diverse cell types. Recent work has shown that SNAP-25 and SNAP-23 are partly localized in sphingolipid/cholesterol-rich lipid raft domains of the plasma membrane and that the integrity of these domains is important for exocytosis. Here, we show that raft localization is mediated by a 36-amino-acid region of SNAP-25 that is also the minimal sequence required for membrane targeting; this domain contains 4 closely spaced cysteine residues that are sites for palmitoylation. Analysis of endogenous levels of SNAP-25 and SNAP-23 present in lipid rafts in PC12 cells revealed that SNAP-23 (54% raft-associated) was almost 3-fold more enriched in rafts when compared with SNAP-25 (20% raft-associated). We report that the increased raft association of SNAP-23 occurs due to the substitution of a highly conserved phenylalanine residue present in SNAP-25 with a cysteine residue. Intriguingly, although the extra cysteine in SNAP-23 enhances its raft association, the phenylalanine at the same position in SNAP-25 acts to repress the raft association of this protein. These different raft-targeting signals within SNAP-25 and SNAP-23 are likely important for fine-tuning the exocytic pathways in which these proteins operate. PMID- 15542598 TI - Proteolysis of non-phosphorylated and phosphorylated tau by thrombin. AB - The microtubule-associated protein tau aggregates intracellularly by unknown mechanisms in Alzheimer's disease and other tauopathies. A contributing factor may be a failure to break down free cytosolic tau, thus creating a surplus for aggregation, although the proteases that degrade tau in brain remain unknown. To address this issue, we prepared cytosolic fractions from five normal human brains and from perfused rat brains and incubated them with or without protease inhibitors. D-Phenylalanyl-L-prolylarginyl chloromethyl ketone, a thrombin specific inhibitor, prevented tau breakdown in these fractions, suggesting that thrombin is a brain protease that processes tau. We next exposed human recombinant tau to purified human thrombin and analyzed the fragments by N terminal sequencing. We found that thrombin proteolyzed tau at multiple arginine and lysine sites. These include Arg(155)-Gly(156), Arg(209)-Ser(210), Arg(230) Thr(231), Lys(257)-Ser(258), and Lys(340)-Ser(341) (numbering according to the longest human tau isoform). Temporally, the initial cleavage occurred at the Arg(155)-Gly(156) bond. Proteolysis of the resultant C-terminal tau fragment then proceeded bidirectionally. When tau was phosphorylated by glycogen synthase kinase-3beta, most of these proteolytic processes were inhibited, except for the first cleavage at the Arg(155)-Gly(156) bond. Furthermore, paired helical filament tau prepared from Alzheimer's disease brain was more resistant to thrombin proteolysis than following dephosphorylation by alkaline phosphatase. The results suggest a possible role for thrombin in proteolysis of tau under physiological and/or pathological conditions in human brains. They are consistent with the hypothesis that phosphorylation of tau inhibits proteolysis by thrombin or other endogenous proteases, leading to aggregation of tau into insoluble fibrils. PMID- 15542599 TI - Genes regulated in neurons undergoing transcription-dependent apoptosis belong to signaling pathways rather than the apoptotic machinery. AB - Neuronal apoptosis has been shown to require de novo RNA/protein synthesis. However, very few genes whose expression is necessary for inducing apoptosis have been identified so far. To systematically identify such genes, we have used genome-scale, long oligonucleotide microarrays and characterized the gene expression profile of cerebellar granule neurons in the early phase of apoptosis elicited by KCl deprivation. We identified 368 significantly differentially expressed genes, including most of the genes previously reported to be transcriptionally regulated in this paradigm. In addition, we identified several hundreds of genes whose transcriptional regulation has never been associated with neuronal apoptosis. We used automated Gene Ontology annotation, analysis of promoter sequences, and statistical tools to characterize these regulations. Although differentially expressed genes included some components of the apoptotic machinery, this functional category was not significantly over-represented among regulated genes. On the other hand, categories related to signal transduction were the most significantly over-represented group. This indicates that the apoptotic machinery is mainly constitutive, whereas molecular pathways that lead to the activation of apoptotic components are transcriptionally regulated. In particular, we show for the first time that signaling pathways known to be involved in the control of neuronal survival are regulated at the transcriptional level and not only by post-translational mechanisms. Moreover, our approach provides insights into novel transcription factors and novel mechanisms, such as the unfolded protein response and cell adhesion, that may contribute to the induction of neuronal apoptosis. PMID- 15542600 TI - Bimodal regulation of the human H1 histamine receptor by G protein-coupled receptor kinase 2. AB - The H1 histamine receptor (H1HR) is a member of the G protein-coupled receptor superfamily and regulates numerous cellular functions through its activation of the G(q/11) subfamily of heterotrimeric G proteins. Although the H1HR has been shown to undergo desensitization in multiple cell types, the mechanisms underlying the regulation of H1HR signaling are poorly defined. To address this issue, we examined the effects of wild type and mutant G protein-coupled receptor kinases (GRKs) on the phosphorylation and signaling of human H1HR in HEK293 cells. Overexpression of GRK2 promoted H1HR phosphorylation in intact HEK293 cells and completely inhibited inositol phosphate production stimulated by H1HR, whereas GRK5 and GRK6 had lesser effects on H1HR phosphorylation and signaling. Interestingly, catalytically inactive GRK2 (GRK2-K220R) also significantly attenuated H1HR-mediated inositol phosphate production, as did an N-terminal fragment of GRK2 previously characterized as a regulator of G protein signaling (RGS) protein for Galpha(q/11). Disruption of this RGS function in holo-GRK2 by mutation (GRK2-D110A) partially reversed the quenching effect of GRK2, whereas deletion of both the kinase activity and RGS function (GRK2-D110A/K220R) effectively relieved the inhibition of inositol phosphate generation. To evaluate the role of endogenous GRKs on H1HR regulation, we used small interfering RNAs to selectively target GRK2 and GRK5, two of the primary GRKs expressed in HEK293 cells. A GRK2-specific small interfering RNA effectively reduced GRK2 expression and resulted in a significant increase in histamine-promoted calcium flux. In contrast, knockdown of GRK5 expression was without effect on H1HR signaling. These findings demonstrate that GRK2 is the principal kinase mediating H1 histamine receptor desensitization in HEK293 cells and suggest that rapid termination of H1HR signaling is mediated by both the kinase activity and RGS function of GRK2. PMID- 15542601 TI - Activation of the epidermal growth factor receptor by respiratory syncytial virus results in increased inflammation and delayed apoptosis. AB - Respiratory syncytial virus (RSV) preferentially infects lung epithelial cells. Infection by RSV leads to an extended inflammatory response, characterized by the release of interleukin-8 (IL-8). Activation of ERK MAP kinase is required for both RSV-induced inflammation and the extended survival of infected cells. In this study, we analyzed the role of the epidermal growth factor receptor (EGFR) in RSV activation of ERK. We demonstrate for the first time that RSV activates EGFR in lung epithelial cells. Activation of EGFR results in increased ERK activity, contributing to both the inflammatory response (IL-8 release) and prolonging the survival of RSV-infected cells. Inhibition of EGFR with siRNA decreased both ERK activation and IL-8 production after RSV. In analyzing the effect of EGFR activation on survival of RSV-infected cells, we found that EGFR activation by RSV resulted in ERK-dependent alterations in the balance of pro- versus anti-apoptotic Bcl2 proteins. RSV altered the balance between pro- and anti-apoptotic Bcl2 proteins (increased BclxL and decreased BimEL) increasing the relative amount of pro-survival proteins. This occurred in an EGFR-dependent manner. This study supports an important role for EGFR activity in the lifespan and inflammatory potential of RSV-infected epithelial cells. PMID- 15542602 TI - UreG, a chaperone in the urease assembly process, is an intrinsically unstructured GTPase that specifically binds Zn2+. AB - Bacillus pasteurii UreG, a chaperone involved in the urease active site assembly, was overexpressed in Escherichia coli BL21(DE3) and purified to homogeneity. The identity of the recombinant protein was confirmed by SDS-PAGE, protein sequencing, and mass spectrometry. A combination of size exclusion chromatography and multiangle and dynamic laser light scattering established that BpUreG is present in solution as a dimer. Analysis of circular dichroism spectra indicated that the protein contains large portions of helices (15%) and strands (29%), whereas NMR spectroscopy indicated the presence of conformational fluxionality of the protein backbone in solution. BpUreG catalyzes the hydrolysis of GTP with a kcat=0.04 min(-1), confirming a role for this class of proteins in coupling energy requirements and nickel incorporation into the urease active site. BpUreG binds two Zn2+ ions per dimer, with a KD=42 +/- 3 microm, and has a 10-fold lower affinity for Ni2+. A structural model for BpUreG was calculated by using threading algorithms. The protein, in the fully folded state, features the typical structural architecture of GTPases, with an open beta-barrel surrounded by alpha-helices and a P-loop at the N terminus. The protein dynamic behavior observed in solution is critically discussed relative to the structural model, using algorithms for disorder predictions. The results suggest that UreG proteins belong to the class of intrinsically unstructured proteins that need the interaction with cofactors or other protein partners to perform their function. It is also proposed that metal ions such as Zn2+ could have important structural roles in the urease activation process. PMID- 15542603 TI - Phosphorylation-independent stabilization of p27kip1 by the phosphoinositide 3 kinase pathway in glioblastoma cells. AB - The PTEN tumor suppressor gene is a frequent target of somatic mutation, particularly in glioblastoma multiform and prostate cancer. The expression of PTEN in PTEN-mutant glioblastoma cells leads to a cell cycle arrest in G(0)/G(1) that is mediated at least partially by increased p27(kip1) levels. Here we show that p27(kip1) is not regulated by transcriptional control but that p27(kip1) protein shows increased stability after inhibition of the phosphoinositide (PI) 3 kinase pathway. Because p27(kip1) protein stability is known to be regulated by phosphorylation, we have examined modifications in the phosphorylation pattern after PI 3-kinase inhibition. Biochemical evidence suggests that p27(kip1) is phosphorylated on several serine residues, including Ser-10 and Ser-178, but that phosphorylation is unaltered by PI 3-kinase activity. This is further confirmed by the inducible expression of p27(kip1) phosphorylation site mutants, suggesting that p27(kip1) is destabilized in a phosphorylation-independent manner by the PI 3-kinase pathway at the G(1)/S transition. PMID- 15542604 TI - Mechanism of chaperone function in small heat shock proteins: dissociation of the HSP27 oligomer is required for recognition and binding of destabilized T4 lysozyme. AB - Mammalian small heat shock proteins (sHSP) form polydisperse and dynamic oligomers that undergo equilibrium subunit exchange. Current models of their chaperone activity hypothesize that recognition and binding of protein non-native states involve changes in the oligomeric state. The equivalent thermodynamic representation is a set of three coupled equilibria that includes the sHSP oligomeric equilibrium, the substrate folding equilibrium, and the equilibrium binding between the sHSP and the substrate non-native states. To test this hypothesis and define the binding-competent oligomeric state of human Hsp27, we have perturbed the two former equilibria and quantitatively determined the consequences on binding. The substrate is a set of T4 lysozyme (T4L) mutants that bind under conditions that favor the folded state over the unfolded state by 10(2)-10(4)-fold. The concentration-dependent oligomer equilibrium of Hsp27 was perturbed by mutations that alter the relative stability of two major oligomeric states including phosphorylation-mimicking mutations that result in the dissociation to a small multimer over a wide range of concentrations. Correlation of binding isotherms with size exclusion chromatography analysis of the Hsp27 oligomer equilibrium demonstrates that the multimer is the binding-competent state. Binding occurs through two modes, each characterized by different affinity and number of binding sites, and results in T4L.Hsp27 complexes of different hydrodynamic properties. Mutants of the Hsp27 phosphorylation mimic that reverse the reduction in oligomer size also reduce the extent of T4L binding. Taken together, these results suggest a central role for the oligomeric equilibrium in regulating the chaperone activity of sHSP. The mutants identify sequence features important for modulating this equilibrium. PMID- 15542605 TI - Tryptophan insertion mutagenesis of liver fatty acid-binding protein: L28W mutant provides important insights into ligand binding and physiological function. AB - Liver fatty acid-binding protein (FABP) binds a variety of non-polar anionic ligands including fatty acids, fatty acyl CoAs, and bile acids. Previously we prepared charge reversal mutants and demonstrated the importance of lysine residues within the portal region in ligand and membrane binding. We have now prepared several tryptophan-containing mutants within the portal region, and one tryptophan at position 28 (L28W) has proved remarkably effective as an intrinsic probe to further study ligand binding. The fluorescence of the L28W mutant was very sensitive to fatty acid and bile acid binding where a large (up to 4-fold) fluorescence enhancement was obtained. In contrast, the binding of oleoyl CoA reduced tryptophan fluorescence. Positive cooperativity for fatty acid binding was observed while detailed information on the orientation of binding of bile acid derivatives was obtained. The ability of bound oleoyl CoA to reduce the fluorescence of L28W provided an opportunity to demonstrate that fatty acyl CoAs can compete with fatty acids for binding to liver FABP under physiological conditions, further highlighting the role of fatty acyl CoAs in modulating FABP function in the cell. PMID- 15542606 TI - Structure-activity relationships of diastereomeric lysine ring size analogs of the antimicrobial peptide gramicidin S: mechanism of action and discrimination between bacterial and animal cell membranes. AB - Structure-activity relationships were examined in seven gramicidin S analogs in which the ring-expanded analog GS14 [cyclo-(VKLKVdYPLKVKLdYP)] is modified by enantiomeric inversions of its lysine residues. The conformation, amphiphilicity, and self-association propensity of these peptides were investigated by circular dichroism spectroscopy and reversed phase high performance liquid chromatography. (31)P nuclear magnetic resonance spectroscopic and dye leakage experiments were performed to evaluate the capacity of these peptides to induce inverse nonlamellar phases in, and to permeabilize phospholipid bilayers; their growth inhibitory activity against the cell wall-less mollicute Acholeplasma laidlawii B was also examined. The amount and stability of beta-sheet structure, effective hydrophobicity, propensity for self-association in water, ability to disrupt the organization of phospholipid bilayers, and ability to inhibit A. laidlawii B growth are strongly correlated with the facial amphiphilicity of these GS14 analogs. Also, the magnitude of the parameters segregate these peptides into three groups, consisting of GS14, the four single inversion analogs, and the two multiple inversion analogs. The capacity of these peptides to differentiate between bacterial and animal cell membranes exhibits a biphasic relationship with peptide amphiphilicity, suggesting that there may only be a narrow range of peptide amphiphilicity within which it is possible to achieve the dual therapeutic requirements of high antibiotic effectiveness and low hemolytic activity. These results were rationalized by considering how the physiochemical properties of these GS14 analogs are likely to be reflected in their partitioning into lipid bilayer membranes. PMID- 15542607 TI - Adhesion stimulates direct PAK1/ERK2 association and leads to ERK-dependent PAK1 Thr212 phosphorylation. AB - The Rac1/Cdc42 effector p21-activated kinase (PAK) is activated by various signaling cascades including receptor-tyrosine kinases and integrins and regulates a number of processes such as cell proliferation and motility. PAK activity has been shown to be required for maximal activation of the canonical Ras/Raf/MEK/ERK Map kinase signaling cascade, likely because of PAK co-activation of Raf and MEK. Herein, we found that adhesion signaling also stimulates an association between PAK1 and ERK1/2. PAK1 and ERK1/2 co-immunoprecipitated from rat aortic smooth muscle cells (SMC) plated on fibronectin, and the two proteins co-localized in membrane ruffles and adhesion complexes following PDGF-BB or sphingosine 1-phosphate treatment, respectively. Far Western analysis demonstrated a direct association between the two proteins, and peptide mapping identified an ERK2 binding site within the autoinhibitory domain of PAK1. Interestingly, deletion of a major ERK binding site in PAK attenuates activation of an ERK-dependent serum-responsive element (SRE)-luciferase reporter gene, indicating that association between PAK and ERK is required to facilitate ERK signaling. We also show that ERK2 phosphorylates PAK1 on Thr(212) in vitro and that Thr(212) is phosphorylated in smooth muscle cells following PDGF-BB treatment in an adhesion- and MEK/ERK-dependent fashion. Expression of a phosphomimic variant, PAK-T212E, does not alter ERK association, but markedly attenuates downstream ERK signaling. Taken together, these data suggest that PAK1 may facilitate ERK signaling by serving as a scaffold to recruit Raf, MEK, and ERK to adhesion complexes, and that subsequent growth factor-stimulated phosphorylation of PAK-Thr(212) by ERK may serve to provide a negative feedback signal to control coordinate activation of ERK by growth factor- and matrix induced signals. PMID- 15542608 TI - Variations on the GFP chromophore: A polypeptide fragmentation within the chromophore revealed in the 2.1-A crystal structure of a nonfluorescent chromoprotein from Anemonia sulcata. AB - We have determined to 2.1 A resolution the crystal structure of a dark state, kindling fluorescent protein isolated from the sea anemone, Anemonia sulcata. The chromophore sequence Met(63)-Tyr(64)-Gly(65) of the A. sulcata chromoprotein was previously proposed to comprise a 6-membered pyrazine-type heterocycle (Martynov, V. I., Savitsky, A. P., Martynova, N. Y., Savitsky, P. A., Lukyanov, K. A., and Lukyanov, S. A. (2001) J. Biol. Chem. 276, 21012-21016). However, our crystallographic data revealed the chromophore to comprise a 5-membered p hydroxybenzylideneimidazolinone moiety that adopts a non-coplanar trans conformation within the interior of the GFP beta-can fold. Unexpectedly, fragmentation of the polypeptide was found to occur within the chromophore moiety, at the bond between Cys(62C) and Met(63N1.) Our structural data reveal that fragmentation of the chromophore represents an intrinsic, autocatalytic step toward the formation of the mature chromophore within the specific GFP-like proteins. PMID- 15542610 TI - A Na+/Cl- -coupled GABA transporter, GAT-1, from Caenorhabditis elegans: structural and functional features, specific expression in GABA-ergic neurons, and involvement in muscle function. AB - GABA functions as an inhibitory neurotransmitter in body muscles and as an excitatory neurotransmitter in enteric muscles in Caenorhabditis elegans. Whereas many of the components of the GABA-ergic neurotransmission in this organism have been identified at the molecular and functional levels, no transporter specific for this neurotransmitter has been identified to date. Here we report on the cloning and functional characterization of a GABA transporter from C. elegans (ceGAT-1) and on the functional relevance of the transporter to the biology of body muscles and enteric muscles. ceGAT-1 is coded by snf-11 gene, a member of the sodium-dependent neurotransmitter symporter gene family in C. elegans. The cloned ceGAT-1 functions as a Na(+)/Cl(-)-coupled high-affinity transporter selective for GABA with a K(t) of approximately 15 microm. The Na(+):Cl(-):GABA stoichiometry for ceGAT-1-mediated transport process is 2:1:1. The transport process is electrogenic as evidenced from GABA-induced inward currents in Xenopus laevis oocytes that express ceGAT-1 heterologously. The transporter is expressed exclusively in GABA-ergic neurons and in two other additional neurons. We also investigated the functional relevance of ceGAT-1 to the biology of body muscles and enteric muscles by ceGAT-1-specific RNA interference (RNAi) in rrf-3 mutant, a strain of C. elegans in which neurons are not refractory to RNAi as in the wild type strain. The down-regulation of ceGAT-1 by RNAi leads to an interesting phenotype associated with altered function of body muscles (as evident from changes in thrashing frequency) and enteric muscles (as evident from the rates of defecation failure) and also with altered sensitivity to aldicarb-induced paralysis. These findings provide unequivocal evidence for a modulatory role of GABA and ceGAT-1 in the biology of cholinergic neurons and in the function of body muscles and enteric muscles in this organism. PMID- 15542609 TI - 15-Hydroxyprostaglandin dehydrogenase is down-regulated in colorectal cancer. AB - Prostaglandin E2 (PGE2) can stimulate tumor progression by modulating several proneoplastic pathways, including proliferation, angiogenesis, cell migration, invasion, and apoptosis. Although steady-state tissue levels of PGE2 stem from relative rates of biosynthesis and breakdown, most reports examining PGE2 have focused solely on the cyclooxygenase-dependent formation of this bioactive lipid. Enzymatic degradation of PGE2 involves the NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH). The present study examined a range of normal tissues in the human and mouse and found high levels of 15-PGDH in the large intestine. By contrast, the expression of 15-PGDH is decreased in several colorectal carcinoma cell lines and in other human malignancies such as breast and lung carcinomas. Consistent with these findings, we observe diminished 15-Pgdh expression in ApcMin+/- mouse adenomas. Enzymatic activity of 15-PGDH correlates with expression levels and the genetic disruption of 15-Pgdh completely blocks production of the urinary PGE2 metabolite. Finally, 15-PGDH expression and activity are significantly down-regulated in human colorectal carcinomas relative to matched normal tissue. In summary, these results suggest a novel tumor suppressive role for 15-PGDH due to loss of expression during colorectal tumor progression. PMID- 15542611 TI - TRPC1-mediated inhibition of 1-methyl-4-phenylpyridinium ion neurotoxicity in human SH-SY5Y neuroblastoma cells. AB - Mammalian homologues of the Drosophila canonical transient receptor potential (TRP) proteins have been implicated to function as plasma membrane Ca(2+) channels. This study examined the role of TRPC1 in human neuroblastoma (SH-SY5Y) cells. SH-SY5Y cells treated with an exogenous neurotoxin, 1-methyl-4 phenylpyridinium ion (MPP(+)) significantly decreased TRPC1 protein levels. Confocal microscopy on SH-SY5Y cells treatment with MPP(+) showed decreased plasma membrane staining of TRPC1. Importantly, overexpression of TRPC1 reduced neurotoxicity induced by MPP(+). MPP(+)-induced alpha-synuclein expression was also suppressed by TRPC1 overexpression. Protection of SH-SY5Y cells against MPP(+) was significantly decreased upon the overexpression of antisense TRPC1 cDNA construct or the addition of a nonspecific transient receptor potential channel blocker lanthanum. Activation of TRPC1 by thapsigargin or carbachol decreased MPP(+) neurotoxicity, which was partially dependent on external Ca(2+). Staining of SH-SY5Y cells with an apoptotic marker (YO-PRO-1) showed that TRPC1 protects SH-SY5Y neuronal cells against apoptosis. Further, TRPC1 overexpression inhibited cytochrome c release and decreased Bax and Apaf-1 protein levels. Interpretation of the above data suggests that reduction in the cell surface expression of TRPC1 following MPP(+) treatment may be involved in dopaminergic neurodegeneration. Furthermore, TRPC1 may inhibit degenerative apoptotic signaling to provide neuroprotection against Parkinson's disease-inducing agents. PMID- 15542612 TI - Reconstitution of GDP-mannose transport activity with purified Leishmania LPG2 protein in liposomes. AB - Activated nucleotide sugars required for the synthesis of glycoconjugates within the secretory pathway of eukaryotes are provided by the action of nucleotide sugar transporters (NSTs). Typically, NSTs are studied in microsomal preparations from wild-type or mutant lines; however, in this setting it can be difficult to assess NST properties because of the presence of glycosyltransferases and other interfering activities. Here we have engineered Leishmania donovani to express high levels of an active LPG2 Golgi GDP-Man transporter bearing a C-terminal polyhistidine tag. The functional LPG2-HIS was solubilized, purified by metal affinity chromatography, and reconstituted into phosphatidylcholine-containing liposomes using polystyrene SM-2 beads. The proteoliposomes exhibited robust GDP Man transport activity with an apparent K(m) of 6.6 mum. Transport activity was enhanced by preloading of GMP and showed specificity for multiple substrates (GDP Ara and GDP-Fuc). In contrast to the activity in crude microsomes, transport was not dependent on the presence of divalent cations. Thus, reconstitution of transport activity using purified LPG2 protein in liposomes provides firm experimental evidence that a single polypeptide is solely required for NST activity and is able to mediate the uptake of multiple substrates. These studies are relevant to the study of NST structure and function in both protozoan parasites as well as their higher eukaryotic hosts. PMID- 15542614 TI - Differences in transductional tropism of adenoviral and lentiviral vectors in the rat brainstem. AB - Adenoviral vectors (AVVs) and lentiviral vectors (LVVs) are highly useful research tools which can be used to investigate the function of specific cell phenotypes in the brain. The transductional tropism of viral vectors has a critical impact upon the transgene expression in different brain areas. This largely depends on the properties of the viral particles, which for AVVs are most commonly analogous to the serotype 5 adenovirus and, in the case of LVVs, are determined by the envelope used for pseudotyping, for example the vesicular stomatitis virus coat (VSVG). We have created a matching set of shuttle plasmids that allow a one-step transfer of an entire expression cassette between the backbones of AVVs and LVVs. This has permitted a fair assessment of the impact of the vector type on tropism for both AVVs and LVVs. Thus, the aims of this study were twofold: (i) to develop and demonstrate the validity of a transgene 'swap' system between AVVs and LVVs; and (ii) using this system, to assess the tropism of AVVs and LVVs for neuronal versus glial cell types. We have constructed AVVs and VSVG-coated LVVs to express monomeric red fluorescent protein (mRFP) driven by the human cytomegalovirus promoter (hCMV). Transgene expression in neurones and glia in the hypoglossal and dorsal vagal motor nuclei of the rat brainstem was compared by determining the colocalization with immunostaining for the neuronal marker NeuN (neuronal nuclear antigen) and the glial marker GFAP (glial fibrillatory acidic protein). We found that 55% of mRFP-expressing cells transduced with AVVs were immunopositive for GFAP, while only 38% were NeuN immunoreactive. In contrast, when the same expression cassette was delivered by VSVG-coated LVVs, the neurone/glia ratio of mRFP expression was reversed with 56% of mRFP-positive cells identified as neurones and 26% as glia. Thus, the present study provides compelling evidence that VSVG-coated LVVs significantly shift transgene expression towards neurones while transduction with AVVs favours glia. PMID- 15542615 TI - Differential cardiovascular responses to stressors in hypertensive and normotensive rats. AB - The aim of this study was to determine to what extent stress-induced cardiovascular responses depend upon rat strain and/or stressor. Spontaneously hypertensive rats (SHRs), Wistar-Kyoto rats (WKYs) and Sprague-Dawley (SD) rats were implanted with telemetry probes in order to measure heart rate and blood pressure changes when exposed to a stressor. The stress protocols employed included handling, air-jet and restraint, where each stressor was repeated over 10 consecutive days. In addition, a heterologous protocol was established whereby the experimental groups having experienced 10 days of air-jet stress were then immediately exposed to 10 consecutive days of restraint. Each stressor caused graded tachycardic and pressor responses in all strains. For all strains, the magnitude and duration of heart rate and blood pressure increases were greatest in the restraint-based protocols while handling and air-jet caused submaximal changes. A comparison between strains indicated that SHRs exhibited prolonged pressor responses to each of the stressor types tested as compared to the normotensive strains. In addition, repeated exposure over 10 days to handling and air-jet in SHRs caused tachycardic and/or pressor responses to adapt to 'normotensive-like' levels. Heterologous restraint stress caused sensitization of cardiovascular responses upon first exposure, predominantly in normotensive strains. Collectively these data show that the magnitude and duration of the tachycardia and pressor responses evoked by the stressors were different within the strains and were also modified by prior experience. In addition, the cardiovascular profiles presented in this study demonstrate that, within each strain, the heart rate response during stress is graded according to the type of stressor encountered. PMID- 15542616 TI - Neuronal gene transfer by baculovirus-derived vectors accommodating a neurone specific promoter. AB - Recombinant baculoviruses have been employed as gene delivery vectors for mammalian cells, including neurones, during recent years. The aim of the current study was to develop a new recombinant baculovirus vector capable of enhancing gene expression in neurones. A hybrid promoter constructed by fusing the enhancer of human cytomegalovirus (CMV) immediately early promoter to the human platelet derived growth factor (PDGF) beta-chain promoter was placed into a baculovirus expression cassette. In cultured neurones, baculovirus vectors containing the hybrid promoter augmented transgene expression up to 100-fold greater than that mediated by titre-matched baculovirus vectors with the PDGF promoter alone. Double immunostaining of tissue sections collected from the striatum and the retina injected with the new baculovirus vector demonstrated its specificity in driving gene expression almost exclusively in neurones, confirming the feasibility of using a tissue-specific promoter in the context of baculovirus vectors to provide cell type-specific transgene expression. The attributes of the new baculovirus vector might have practical implications for gene therapy in the nervous system. PMID- 15542617 TI - Optimizing regulatable gene expression using adenoviral vectors. AB - Inducible gene expression systems have typically encountered limitations, such as pleitropic effects of the inducer, basal leakiness, toxicity of inducing agents and low levels of expression. However, recently non-toxic, tightly regulated control of transgene expression has been reported for several systems, the most frequently cited being the tetracycline gene control system. We have found that the individual components of the Tet system [the Tet transactivators and tetracycline responsive element (TRE)] function optimally to control gene expression when they are incorporated into separate adenoviral vectors. Furthermore, incorporation of the Woodchuck hepatitis virus post-transcriptional enhancer (WPRE) allows a dual vector Tet-regulatable Ad system to be used at very low titres (2 x 10(4)) that elicit a minimal inflammatory response, with no loss of transgene expression or ability to regulate transgene expression. This and similar regulatable systems will benefit studies investigating neuronal gene function and those seeking to develop effective neuronal gene therapy strategies. PMID- 15542618 TI - Targeting specific neuronal populations using adeno- and lentiviral vectors: applications for imaging and studies of cell function. AB - We employ viral vectors to address questions related to the function of specific types of neurones in the central control of blood pressure. Adenoviral vectors (AVVs) or lentiviral vectors (LVVs) can be used to visualize specifically living GABAergic or noradrenergic (NAergic) neurones or to interfere with intracellular signalling within these cell types. Here, we review recent in vitro, in situ and in vivo applications of these vectors in the rat brainstem as performed in our laboratories. In organotypic slice cultures prepared from defined cardiovascular brainstem areas, viral vectors were used to study the electrophysiological properties, intracellular signalling and gene expression in selected neuronal phenotypes. In vivo, vectors were microinjected into brainstem nuclei to inhibit specific aspects of cell signalling by expression of dominant negative proteins, for example. Outcomes for cardiovascular control were measured either acutely in situ or chronically in vivo with radio telemetry in freely moving rats. We showed that AVVs and LVVs have distinct properties that need to be considered prior to their application. For example, LVVs can be manufactured very quickly, have no immunogenicity and can be pseudotyped to display higher tropism for neurones than glia. However, comparatively lower production yields of LVVs may limit their use for some types of applications. In contrast, AVVs require a lengthy construction period, are easy to amplify to high yields at moderate cost but may trigger an immune response when used at high titres in vivo. These features make AVVs particularly suitable for in vitro applications. As the two vector types complement each other in several ways we generated a shuttle system that simplifies transfer of transgene cassettes between the backbones of AVVs and LVVs. Thus, AVVs and LVVs are powerful experimental tools that can be used in a variety of experimental designs in vivo, in situ and in vitro. PMID- 15542619 TI - Promoters and serotypes: targeting of adeno-associated virus vectors for gene transfer in the rat central nervous system in vitro and in vivo. AB - The brain parenchyma consists of several different cell types, such as neurones, astrocytes, microglia, oligodendroglia and epithelial cells, which are morphologically and functionally intermingled in highly complex three-dimensional structures. These different cell types are also present in cultures of brain cells prepared to serve as model systems of CNS physiology. Gene transfer, either in a therapeutic attempt or in basic research, is a fascinating and promising tool to manipulate both the complex physiology of the brain and that of isolated neuronal cells. Viral vectors based on the parvovirus, adeno-associated virus (AAV), have emerged as powerful transgene delivery vehicles. Here we describe highly efficient targeting of AAV vectors to either neurones or astrocytes in cultured primary brain cell cultures. We also show that transcriptional targeting can be achieved by the use of small promoters, significantly boosting the transgene capacity of the recombinant viral genome. However, we also demonstrate that successful targeting of a vector in vitro does not necessarily imply that the same targeting works in the adult brain. Cross-packaging the AAV-2 genome in capsids of other serotypes adds additional benefits to this vector system. In the brain, the serotype-5 capsid allows for drastically increased spread of the recombinant vector as compared to the serotype-2 capsid. Finally, we emphasize the optimal targeting approach, in which the natural tropism of a vector for a specific cell type is employed. Taken together, these data demonstrate the flexibility which AAV-based vector systems offer in physiological research. PMID- 15542620 TI - Alphaviral cytotoxicity and its implication in vector development. AB - A great variety of viruses have been engineered to serve as expression vectors. Among them, the alphaviruses Semliki Forest virus and Sindbis virus represent promising tools for heterologous gene expression in a wide variety of host cells. Several applications have already been described in neurobiological studies, in gene therapy, for vaccine development and in cancer therapy. Both viruses trigger stress pathways in the cells they infect, sometimes culminating in the death of the host. This inherent property is either an advantage or a drawback, depending on the type of application. This review covers the development and applications of alphavirus vectors and, as our work has been mainly with Semliki Forest virus, we have focused on this virus with special emphasis on how the understanding of Semliki Forest virus cytotoxicity enables it to be manipulated and used. PMID- 15542621 TI - Cell-selective viral gene delivery vectors for the vasculature. AB - Clinical gene therapy for cardiovascular disease remains achievable. To date, however, preclinical studies and clinical trials have highlighted shortfalls in viral gene delivery to vascular cells. These include poor efficiency, poor target tissue selectivity, the presence of pre-existing neutralizing antibodies and immunogenicity generated by the host to vectors such as adenovirus. These important issues require careful consideration when applying viral vectors for gene therapy. Each delivery vector requires precise optimization and tailoring for each disease application since parameters relating to vector : tissue exposure time, route of delivery and target cell type vary considerably. Optimization can be achieved through modification of the structure of the virus capsid proteins and expression cassette to generate vectors that are highly selective and efficient for target cell binding and entry as well as instilling transcriptional control and/or longevity on transgene expression. This ultimately will improve the efficacy and toxicity profiles of gene delivery vectors and has become a very important area in gene therapy. Here, we review recent advances in the targeting of viral gene delivery vectors to the vasculature. PMID- 15542622 TI - Differential mechanisms of antianalgesia induced by endomorphin-1 and endomorphin 2 in the ventral periaqueductal gray of the rat. AB - The effects of pretreatment with endomorphin-1 (EM-1) and endomorphin-2 (EM-2) given into the ventral periaqueductal gray (vPAG) to induce antianalgesia against the tail-flick (TF) inhibition produced by morphine given into the vPAG were studied in rats. Pretreatment with EM-1 (3.5-28 nmol) given into vPAG for 45 min dose-dependently attenuated the TF inhibition produced by morphine (9 nmol) given into vPAG. Similarly, pretreatment with EM-2 (1.7-7.0 nmol) for 45 min also attenuated the TF inhibition induced by morphine; however, a high dose of EM-2 (14 nmol) did not attenuate the morphine-produced TF inhibition. The attenuation of morphine-produced TF inhibition induced by EM-2 or EM-1 pretreatment was blocked by pretreatment with mu-opioid antagonist (-)-naloxone (55 pmol) but not nonopioid (+)-naloxone (55 pmol). However, pretreatment with a morphine-6beta glucuronide-sensitive mu-opioid receptor antagonist 3-methoxynaltrexone (6.4 pmol) selectively blocked EM-2- but not EM-1-induced antianalgesia. Pretreatment with dynorphin A(1-17) antiserum reversed only EM-2- but not EM-1-induced antianalgesia. Pretreatment with antiserum against beta-endorphin, [Met(5)]enkephalin, [Leu(5)]enkephalin, substance P or cholecystokinin, or with delta-opioid receptor antagonist naltrindole (2.2 nmol) or kappa-opioid receptor antagonist norbinaltorphimine (6.6 nmol) did not affect EM-2-induced antianalgesia. It is concluded that EM-2 selectively releases dynorphin A(1-17) by stimulation of a novel subtype of mu-opioid receptor, tentatively designated as mu(3) in the vPAG to induce antianalgesia, whereas the antianalgesia induced by EM-1 is mediated by the stimulation of another subtype of mu(1)- or mu(2) opioid receptor. PMID- 15542623 TI - Nicotinic acetylcholine receptor-mediated [3H]dopamine release from hippocampus. AB - The mechanism of nicotinic acetylcholine receptor (nAChR)-induced hippocampal dopamine (DA) release was investigated using rat hippocampal slices. nAChRs involved in hippocampal DA and norepinephrine (NE) release were investigated using prototypical agonists and antagonists and several relatively novel compounds: ABT-594 [(R)-5-(2-azetidinylmethoxy)-2-chloropyridine], (+/-)-UB-165 [(2-chloro-5-pyridyl)-9-azabicyclo [4.2.1]non2-ene], and MG 624 [N,N,N-triethyl-2 [4-(2 phenylethenyl)phenoxy]-ethanaminium iodine]. (+/-)-Epibatidine, (+/-)-UB 165, anatoxin-a, ABT-594, (-)-nicotine, 1,1-dimethyl-4-phenyl-piperazinium iodide, and (-)-cytisine (in decreasing order of potency) evoked [(3)H]DA release in a mecamylamine-sensitive manner. Aside from (+/-)-UB-165, all the agonists displayed full efficacy relative to 100 microM (-)-nicotine in [(3)H]DA release. In contrast, (+/-)-UB-165 was a partial agonist, evoking 58% of 100 microM (-) nicotine response. Mecamylamine, MG 624, hexamethonium, d-tubocurare, and dihydro beta-erythroidine (in decreasing order of potency), but not alpha-conotoxin-MII, methyllycaconitine, alpha-conotoxin-ImI, or alpha-bungarotoxin, attenuated 100 microM (-)-nicotine-evoked [(3)H]DA release in a concentration-dependent manner. (+/-)-UB-165, ABT-594, and MG 624 exhibited different pharmacologic profiles in the [(3)H]NE release assay when compared with their effect on [(3)H]DA release. ABT-594 was 4.5-fold more potent, and (+/-)-UB-165 was a full agonist in contrast to its partial agonism in [(3)H]DA release. MG 624 potently and completely blocked NE release evoked by 100 microM (-)-nicotine and 10 microM (+/-)-UB-165, whereas it only partially inhibited (-)-nicotine-evoked [(3)H]DA release. In conclusion, we provide evidence that [(3)H]DA can be evoked from the hippocampus and that the pharmacologic profile for nAChR-evoked hippocampal [(3)H]DA release suggests the involvement of alpha3beta4(*) and at least one other nAChR subtype, thus distinguishing it from that of nAChR-evoked hippocampal [(3)H]NE release. PMID- 15542624 TI - Dopamine uptake inhibitor-induced rotation in 6-hydroxydopamine-lesioned rats involves both D1 and D2 receptors but is modulated through 5-hydroxytryptamine and noradrenaline receptors. AB - Dopamine uptake inhibitors may provide a means of sustaining endogenous and exogenous striatal dopamine levels in Parkinson's disease, but most are not selective and also inhibit the noradrenaline and 5-hydroxytryptamine (5-HT) transporters. To determine the involvement of the individual monoamine transporters in the production of motor activity, the effect of the nonselective monoamine uptake inhibitor BTS 74 398 1-([1-(3,4-dichlorophenyl)cyclobutyl]-2-(3 diaminethylaminopropylthio) ethanone monocitrate) and the selective dopamine, GBR 12909 [1-(2-(bis-(4-fluorphenyl)-methyl)ethyl)-4-(3-phenylpropyl)piperazine) dihydrochloride], noradrenaline (nisoxetine), and 5-HT (fluvoxamine) reuptake inhibitors on circling in the unilateral 6-hydroxydopamine-lesioned rat was investigated. GBR 12909 induced ipsilateral circling, but fluvoxamine and nisoxetine were without effect. However, when administered with GBR 12909, fluvoxamine enhanced rotation, whereas nisoxetine had no effect. The results suggest that 5-HT, but not noradrenaline, reuptake inhibition facilitates dopamine-mediated motor activity. To test this hypothesis, BTS 74 398 was administered in combination with selective dopamine, 5-HT, and noradrenaline receptor antagonists. Both D(1) and D(2) receptor antagonists, SCH 23390 [R(+)-7 chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine] and raclopride, inhibited BTS 74 398-induced circling. In contrast, the 5-HT(1A) 5 HT(1A/B) antagonists, WAY 100635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl] N-2-pyridinyl-cyclohexane-carboxamide maleate) and pindolol, and the 5-HT(2A) antagonist, ketanserin, had no effect. The nonspecific 5-HT((1/2)) antagonists, methysergide and metergoline, and the specific 5-HT(2C) antagonist, N desmethylclozapine, enhanced BTS 74 398-induced circling, as did the alpha(2) adrenoceptor antagonist idazoxan. Overall, the data suggest that inhibition of the 5-HT and noradrenaline transporters modulate dopamine uptake inhibitor mediated motor activity. However, the mechanism of this interaction is complex, involving opposing effects of noradrenaline and 5-HT agonism and antagonism. PMID- 15542625 TI - The enigmatic X gene of hepatitis B virus. PMID- 15542627 TI - Resistance to vesicular stomatitis virus infection requires a functional cross talk between the eukaryotic translation initiation factor 2alpha kinases PERK and PKR. AB - Phosphorylation of the alpha (alpha) subunit of the eukaryotic translation initiation factor 2 (eIF2) leads to the inhibition of protein synthesis in response to diverse stress conditions, including viral infection. The eIF2alpha kinase PKR has been shown to play an essential role against vesicular stomatitis virus (VSV) infection. We demonstrate here that another eIF2alpha kinase, the endoplasmic reticulum-resident protein kinase PERK, contributes to cellular resistance to VSV infection. We demonstrate that mouse embryonic fibroblasts (MEFs) from PERK(-/-) mice are more susceptible to VSV-mediated apoptosis than PERK(+/+) MEFs. The higher replication capacity of VSV in PERK(-/-) MEFs results from their inability to attenuate viral protein synthesis due to an impaired eIF2alpha phosphorylation. We also show that VSV-infected PERK(-/-) MEFs are unable to fully activate PKR, suggesting a cross talk between the two eIF2alpha kinases in virus-infected cells. These findings further implicate PERK in virus infection, and provide evidence that the antiviral and antiapoptotic roles of PERK are mediated, at least in part, via the activation of PKR. PMID- 15542626 TI - Class II integrase mutants with changes in putative nuclear localization signals are primarily blocked at a postnuclear entry step of human immunodeficiency virus type 1 replication. AB - Integrase has been implicated in human immunodeficiency virus type 1 (HIV-1) nuclear import. Integrase analyses, however, can be complicated by the pleiotropic nature of mutations: whereas class I mutants are integration defective, class II mutants display additional assembly and/or reverse transcription defects. We previously determined that HIV-1(V165A), originally reported as defective for nuclear import, was a class II mutant. Here we analyzed mutants containing changes in other putative nuclear localization signals, including (186)KRK(188)/(211)KELQKQITK(219) and Cys-130. Previous work established HIV-1(K186Q), HIV-1(Q214L/Q216L), and HIV-1(C130G) as replication defective, but phenotypic classification was unclear and nuclear import in nondividing cells was not addressed. Consistent with previous reports, most of the bipartite mutants studied here were replication defective. These mutants as well as HIV-1(V165A) synthesized reduced cDNA levels, but a normal fraction of mutant cDNA localized to dividing and nondividing cell nuclei. Somewhat surprisingly, recombinant class II mutant proteins were catalytically active, and class II Vpr-integrase fusion proteins efficiently complemented class I mutant virus. Since a class I Vpr-integrase mutant efficiently complemented class II mutant viruses under conditions in which class II Vpr-integrases failed to function, we conclude that classes I and II define two distinct complementation groups and suggest that class II mutants are primarily defective at a postnuclear entry step of HIV-1 replication. HIV-1(C130G) was also defective for reverse transcription, but Vpr-integrase(C130G) did not efficiently complement class I mutant HIV-1. Since HIV-1(C130A) grew like the wild type, we conclude that Cys 130 is not essential for replication and speculate that perturbation of integrase structure contributed to the pleiotropic HIV-1(C130G) phenotype. PMID- 15542628 TI - Conserved methylation patterns of human papillomavirus type 16 DNA in asymptomatic infection and cervical neoplasia. AB - DNA methylation contributes to the chromatin conformation that represses transcription of human papillomavirus type16 (HPV-16), which is prevalent in the etiology of cervical carcinoma. In an effort to clarify the role of this phenomenon in the regulation and carcinogenicity of HPV-16, 115 clinical samples were studied to establish the methylation patterns of the 19 CpG dinucleotides within the long control region and part of the L1 gene by bisulfite modification, PCR amplification, DNA cloning, and sequencing. We observed major heterogeneities between clones from different samples as well as between clones from individual samples. The methylation frequency of CpGs was measured at 14.5%. In addition, 0.21 and 0.23%, respectively, of the CpA and CpT sites, indicators of de novo methylation, were methylated. Methylation frequencies exceeded 30% in the CpGs overlapping with the L1 gene and were about 10% for most other positions. A CpG site located in the linker between two nucleosomes positioned over the enhancer and promoter of HPV-16 had minimal methylation. This region forms part of the HPV replication origin and is close to binding sites of master-regulators of transcription during epithelial differentiation. Methylation of most sites was highest in carcinomas, possibly due to tandem repetition and chromosomal integration of HPV-16 DNA. Methylation was lowest in dysplasia, likely reflecting the transcriptional activity in these infections. Our data document the efficient targeting of HPV genomes by the epithelial methylation machinery, possibly as a cellular defense mechanism, and suggest involvement of methylation in HPV oncogene expression and the early-late switch. PMID- 15542629 TI - Nef associates with p21-activated kinase 2 in a p21-GTPase-dependent dynamic activation complex within lipid rafts. AB - We have previously reported that Nef specifically interacts with a small but highly active subpopulation of p21-activated kinase 2 (PAK2). Here we show that this is due to a transient association of Nef with a PAK2 activation complex within a detergent-insoluble membrane compartment containing the lipid raft marker GM1. The low abundance of this Nef-associated kinase (NAK) complex was found to be due to an autoregulatory mechanism. Although activation of PAK2 was required for assembly of the NAK complex, catalytic activity of PAK2 also promoted dissociation of this complex. Testing different constitutively active PAK2 mutants indicated that the conformation associated with p21-mediated activation rather than kinase activity per se was required for PAK2 to become NAK. Although association with PAK2 is one of the most conserved properties of Nef, we found that the ability to stimulate PAK2 activity differed markedly among divergent Nef alleles, suggesting that PAK2 association and activation are distinct functions of Nef. However, mutations introduced into the p21-binding domain of PAK2 revealed that p21-GTPases are involved in both of these Nef functions and, in addition to promoting PAK2 activation, also help to physically stabilize the NAK complex. PMID- 15542630 TI - The zinc finger antiviral protein directly binds to specific viral mRNAs through the CCCH zinc finger motifs. AB - The zinc finger antiviral protein (ZAP) is a recently isolated host antiviral factor. It specifically inhibits the replication of Moloney murine leukemia virus (MLV) and Sindbis virus (SIN) by preventing the accumulation of viral RNA in the cytoplasm. For this report, we mapped the viral sequences that are sensitive to ZAP inhibition. The viral sequences were cloned into a luciferase reporter and analyzed for the ability to mediate ZAP-dependent destabilization of the reporter. The sensitive sequence in MLV was mapped to the 3' long terminal repeat; the sensitive sequences in SIN were mapped to multiple fragments. The fragment of SIN that displayed the highest destabilizing activity was further analyzed by deletion mutagenesis for the minimal sequence that retained the activity. This led to the identification of a fragment of 653 nucleotides. Any further deletion of this fragment resulted in significantly lower activity. We provide evidence that ZAP directly binds to the active but not the inactive fragments. The CCCH zinc finger motifs of ZAP play important roles in RNA binding and antiviral activity. Disruption of the second and fourth zinc fingers abolished ZAP's activity, whereas disruption of the first and third fingers just slightly lowered its activity. PMID- 15542631 TI - Role of the proximal enhancer of the major immediate-early promoter in human cytomegalovirus replication. AB - The human cytomegalovirus (CMV) enhancer has a distal component (positions -550 to -300) and a proximal component (-300 to -39) relative to the transcription start site (+1) of the major immediate-early (MIE) promoter. Without the distal enhancer, human CMV replicates slower and has a small-plaque phenotype. We determined the sequence requirements of the proximal enhancer by making 5'-end deletions to positions -223, -173, -116, -67, and -39. Even though recombinant virus with the proximal enhancer deleted to -39 has the minimal TATA box containing MIE promoter element, it cannot replicate independently in human fibroblast cells. Recombinant virus with a deletion to -67 has an Sp-1 transcription factor binding site which may represent a minimal enhancer element for recombinant virus replication in human fibroblast cells. Although recombinant virus with a deletion to -223 replicates to titers at least 100-fold less than that of the wild-type virus, it replicates to titers 8-fold higher than that of recombinant virus with a deletion to -173 and 20-fold higher than that of virus with a deletion to -67. Recombinant virus with a deletion to -173 replicates more efficiently than that with a deletion to -116. There was a direct correlation between the level of infectious virus replication and time after infection, amount of MIE gene transcription, MIE and early viral protein synthesis, and viral DNA synthesis. The extent of the proximal enhancer determines the efficiency of viral replication. PMID- 15542632 TI - Target cell cyclophilin A modulates human immunodeficiency virus type 1 infectivity. AB - The peptidyl-prolyl isomerase cyclophilin A (CypA) increases the kinetics by which human immunodeficiency virus type 1 (HIV-1) spreads in tissue culture. This was conclusively demonstrated by gene targeting in human CD4(+) T cells, but the role of CypA in HIV-1 replication remains unknown. Though CypA binds to mature HIV-1 capsid protein (CA), it is also incorporated into nascent HIV-1 virions via interaction with the CA domain of the Gag polyprotein. These findings raised the possibility that CypA might act at multiple steps of the retroviral life cycle. Disruption of the CA-CypA interaction, either by the competitive inhibitor cyclosporine (CsA) or by mutation of CA residue G89 or P90, suggested that producer cell CypA was required for full virion infectivity. However, recent studies indicate that CypA within the target cell regulates HIV-1 infectivity by modulating Ref1- or Lv1-mediated restriction. To examine the relative contribution to HIV-1 replication of producer cell CypA and target cell CypA, we exploited multiple tools that disrupt the HIV-1 CA-CypA interaction. These tools included the drugs CsA, MeIle(4)-CsA, and Sanglifehrin; CA mutants exhibiting decreased affinity for CypA or altered CypA dependence; HeLa cells with CypA knockdown by RNA interference; and Jurkat T cells homozygous for a deletion of the gene encoding CypA. Our results clearly demonstrate that target cell CypA, and not producer cell CypA, is important for HIV-1 CA-mediated function. Inhibition of HIV-1 infectivity resulting from virion production in the presence of CsA occurs independently of the CA-CypA interaction or even of CypA. PMID- 15542633 TI - Conventional protein kinase C inhibition prevents alpha interferon-mediated hepatitis C virus replicon clearance by impairing STAT activation. AB - Hepatitis C virus (HCV) has evolved complex strategies to evade host immune responses and establish chronic infection. The only treatment available for HCV infections, alpha interferon (IFN-alpha), is effective in a limited percentage of patients. The mechanisms by which IFN-alpha interferes with the HCV life cycle and the reasons for limited effectiveness of IFN-alpha therapy have not yet been fully elucidated. Using a cell-based HCV replication system and specific kinase inhibitors, we examined the role played by various signaling pathways in the IFN alpha-mediated HCV clearance. We reported that conventional protein kinase C (cPKC) activity is important for the effectiveness of IFN-alpha treatment. In cells treated with a cPKC-specific inhibitor, IFN-alpha failed to induce an efficient HCV RNA degradation. The lack of cPKC activity leads to a broad reduction of IFN-alpha-stimulated gene expression due to a significant impairment of STAT1 and STAT3 tyrosine phosphorylation. Thus, modulation of cPKC function by either host or viral factors could influence the positive outcome of IFN-alpha mediated antiviral therapies. PMID- 15542634 TI - Multiple genotypes of influenza B virus circulated between 1979 and 2003. AB - The segmented genome of influenza B virus allows exchange of gene segments between cocirculating strains. Through this process of reassortment, diversity is generated by the mixing of genes between viruses that differ in one or more gene segments. Phylogenetic and evolutionary analyses of all 11 genes of 31 influenza B viruses isolated from 1979 to 2003 were used to study the evolution of whole genomes. All 11 genes diverged into two new lineages prior to 1987. All genes except the NS1 gene were undergoing linear evolution, although the rate of evolution and the degree to which nucleotide changes translated into amino acid changes varied between lineages and by gene. Frequent reassortment generated 14 different genotypes distinct from the gene constellation of viruses circulating prior to 1979. Multiple genotypes cocirculated in some locations, and a sequence of reassortment events over time could not be established. The surprising diversity of the viruses, unrestricted mixing of lineages, and lack of evidence for coevolution of gene segments do not support the hypothesis that the reassortment process is driven by selection for functional differences. PMID- 15542635 TI - Identification of cellular cofactors for human immunodeficiency virus replication via a ribozyme-based genomics approach. AB - Ribozymes are small, catalytic RNA molecules that can be engineered to down regulate gene expression by cleaving specific mRNA. Here we report the selection of hairpin ribozymes that inhibit human immunodeficiency virus (HIV) replication from a combinatorial ribozyme library. We identified a total of 17 effective ribozymes, each capable of inhibiting HIV infection of human CD4(+) cells. These ribozymes target diverse steps of the viral replication cycle, ranging from entry to transcription. One ribozyme suppressed HIV integration and transcription by inhibiting the expression of the Ku80 subunit of the DNA-activated protein kinase. Another ribozyme specifically inhibited long terminal repeat transactivation, while two additional ones blocked a step that can be bypassed by vesicular stomatitis virus G-protein pseudotyping. The function of Ku80 in HIV replication and its mechanism of action were further confirmed using short interfering RNA. Identification of the gene targets of these and other selected ribozymes may reveal novel therapeutic targets for combating HIV infection. PMID- 15542636 TI - Coexpression of hepatitis C virus E1 and E2 chimeric envelope glycoproteins displays separable ligand sensitivity and increases pseudotype infectious titer. AB - We have previously reported that a pseudotype virus generated by reconstitution of hepatitis C virus (HCV) chimeric envelope glycoprotein E1-G or E2-G on the surface of a temperature-sensitive mutant of vesicular stomatitis virus (VSVts045) interacts independently with mammalian cells to initiate infection. Here, we examined whether coexpression of both of the envelope glycoproteins on pseudotype particles would augment virus infectivity and/or alter the functional properties of the individual subunits. Stable transfectants of baby hamster kidney (BHK) epithelial cells expressing either one or both of the chimeric envelope glycoproteins of HCV on the cell surface were generated. The infectious titer of the VSV pseudotype, derived from a stable cell line incorporating both of the chimeric glycoproteins of HCV, was approximately 4- to 5-fold higher than that of a pseudotype bearing E1-G alone or approximately 25- to 30-fold higher than that of E2-G alone when assayed with a number of mammalian cell lines. Further studies suggested that that the E1-G/E2-G or E2-G pseudotype was more sensitive to the inhibitory effect of heparin than the E1-G pseudotype. Treatment of the E1-G/E2-G pseudotype with a negatively charged sulfated sialyl lipid (NMSO3) displayed a approximately 4-fold-higher sensitivity to neutralization than pseudotypes with either of the two individual glycoproteins. In contrast, VSVts045, used as a backbone for the generation of pseudotypes, displayed at least 20-fold-higher sensitivity to NMSO3-mediated inhibition of virus plaque formation. The effect of low-density lipoprotein on the E1-G pseudotype was greater than that apparent for the E1-G/E2-G pseudotype. The treatment of cells with monoclonal antibodies to CD81 displayed an inhibitory effect upon the pseudotype with E1-G/E2-G or with E2-G alone. Taken together, our results indicate that the HCV E1 and E2 glycoproteins have separable functional properties and that the presence of these two envelope glycoproteins on VSV/HCV pseudotype particles increases infectious titer. PMID- 15542637 TI - Identification of Epstein-Barr virus RK-BARF0-interacting proteins and characterization of expression pattern. AB - The Epstein-Barr virus (EBV) BamHI A transcripts are a family of transcripts that are differentially spliced and can be detected in multiple EBV-associated malignancies. Several of the transcripts may encode proteins. One transcript of interest, RK-BARF0, is proposed to encode a 279-amino-acid protein with a possible endoplasmic reticulum-targeting sequence. In this study, the properties of RK-BARF0 were examined through identification of cellular-interacting proteins through yeast two-hybrid analysis and characterization of its expression in EBV infected cells and tumors. In addition to the interaction previously identified with cellular Notch, it was determined that RK-BARF0 also bound cellular human I mfa domain-containing protein (HIC), epithelin, and scramblase. An interaction between RK-BARF0 and Notch or epithelin induced proteasome-dependent degradation of Notch and epithelin but not of HIC or scramblase. Low levels of endogenous Notch expression in EBV-positive cell lines may correlate with RK-BARF0 expression. However, a screen of EBV-positive cell lines and tumors with an affinity-purified alpha-RK-BARF0 antiserum did not consistently detect RK-BARF0. These data suggest that while RK-BARF0 may have important cellular functions during EBV infection, and while the phenotype of EBV-positive cells suggest its expression, RK-BARF0 levels may be too low to detect. PMID- 15542638 TI - A cyclin-binding motif within the amino-terminal homology domain of EBNA3C binds cyclin A and modulates cyclin A-dependent kinase activity in Epstein-Barr virus infected cells. AB - The Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA3C) is a virus-encoded latent antigen essential for primary B-cell transformation. In this report we demonstrate that although the carboxy terminus of EBNA3C predominantly regulates cyclin A-dependent kinase activity, the region of greatest affinity for cyclin A lies within the EBNA3 amino-terminal homology domain of EBNA3C. Detailed mapping studies employing both in vitro binding assays and coimmunoprecipitation experiments implicated a small region of EBNA3C, amino acids 130 to 159 within the EBNA3 homology domain, as having the greatest affinity for cyclin A. The EBNA3 homology domain has the highest degree of amino acid similarity (approximately 30%) between the EBNA3 proteins, and, indeed, EBNA3B, but not EBNA3A, showed binding activity with cyclin A. We also show that EBNA3C binds to the alpha1 helix of the highly conserved mammalian cyclin box, with cyclin A amino acids 206 to 226 required for strong binding to EBNA3C amino acids 130 to 159. Interestingly, EBNA3C also bound human cyclins D1 and E in vitro, although the affinity was approximately 30% of that seen for cyclin A. Previously it was demonstrated that full-length EBNA3C rescues p27-mediated suppression of cyclin A dependent kinase activity (J. S. Knight and E. S. Robertson, J. Virol. 78:1981 1991, 2004). It was also demonstrated that the carboxy terminus of EBNA3C recapitulates this phenotype. Surprisingly, the amino terminus of EBNA3C with the highest affinity for cyclin A was unable to rescue p27 suppression of kinase activity and actually downregulates cyclin A activity when introduced into EBV infected cells. The data presented here suggests that the amino terminus of EBNA3C may play an important role in recruiting cyclin A complexes, while the carboxy terminus of EBNA3C is necessary for the functional modulation of cyclin A complex kinase activity. PMID- 15542639 TI - Adenovirus VA1 noncoding RNA can inhibit small interfering RNA and MicroRNA biogenesis. AB - Although inhibition of RNA interference (RNAi) by plant virus proteins has been shown to enhance viral replication and pathogenesis in plants, no viral gene product has as yet been shown to inhibit RNAi in vertebrate cells. Here, we present evidence demonstrating that the highly structured approximately 160 nucleotide adenoviral VA1 noncoding RNA can inhibit RNAi at physiological levels of expression. VA1, which is expressed at very high levels in adenovirus-infected cells, potently inhibited RNAi induced by short hairpin RNAs (shRNAs) or human microRNA precursors but did not affect RNAi induced by artificial short interfering RNA duplexes. Inhibition appeared to be due both to inhibition of nuclear export of shRNA or premicro-RNA precursors, competition for the Exportin 5 nuclear export factor, and inhibition of Dicer function by direct binding of Dicer. Together, these data argue that adenovirus infection can result in inhibition of RNAi and identify VA1 RNA as the first viral gene product able to inhibit RNAi in human cells. PMID- 15542640 TI - Recombinant human Metapneumovirus lacking the small hydrophobic SH and/or attachment G glycoprotein: deletion of G yields a promising vaccine candidate. AB - Human metapneumovirus (HMPV) has recently been identified as a significant cause of serious respiratory tract disease in humans. In particular, the emerging information on the contribution of HMPV to pediatric respiratory tract disease suggests that it will be important to develop a vaccine against this virus for use in conjunction with those being developed for human respiratory syncytial virus and the human parainfluenza viruses. A recently described reverse genetic system (S. Biacchesi, M. H. Skiadopoulos, K. C. Tran, B. R. Murphy, P. L. Collins, and U. J. Buchholz, Virology 321:247-259, 2004) was used to generate recombinant HMPVs (rHMPVs) that lack the G gene, the SH gene, or both. The DeltaSH, DeltaG, and DeltaSH/G deletion mutants were readily recovered and were found to replicate efficiently during multicycle growth in cell culture. Thus, the SH and G proteins are not essential for growth in cell culture. Apart from the absence of the deleted protein(s), the virions produced by the gene deletion mutants were similar by protein yield and gel electrophoresis protein profile to wild-type HMPV. When administered intranasally to hamsters, the DeltaG and DeltaSH/G mutants replicated in both the upper and lower respiratory tracts, showing that HMPV containing F as the sole viral surface protein is competent for replication in vivo. However, both viruses were at least 40-fold and 600-fold restricted in replication in the lower and upper respiratory tract, respectively, compared to wild-type rHMPV. They also induced high titers of HMPV-neutralizing serum antibodies and conferred complete protection against replication of wild type HMPV challenge virus in the lungs. Surprisingly, G is dispensable for protection, and the DeltaG and DeltaSH/G viruses represent promising vaccine candidates. In contrast, DeltaSH replicated somewhat more efficiently in hamster lungs compared to wild-type rHMPV (20-fold increase on day 5 postinfection). This indicates that SH is completely dispensable in vivo and that its deletion does not confer an attenuating effect, at least in this rodent model. PMID- 15542641 TI - Requirement of Sur2 for efficient replication of mouse adenovirus type 1. AB - Mouse adenovirus type 1 (MAV-1) early region 1A (E1A) encodes a virulence gene in viral infection of mice. To broaden our understanding of the functions of E1A in MAV-1 pathogenesis, an unbiased experimental approach, glutathione S-transferase (GST) pulldown, was used to screen for cellular proteins that interact with E1A protein. We identified mouse Sur2, a subunit of Mediator complex, as a protein that binds to MAV-1 E1A. The interaction between Sur2 and MAV-1 E1A was confirmed in virus-infected cells. Conserved region 3 (CR3) of MAV-1 E1A was mapped as the region required for Sur2-E1A interaction, as is the case for human adenovirus E1A. Although it has been proposed that human adenovirus E1A recruits the Mediator complex to transactivate transcription of viral early genes, Sur2 function in adenovirus replication has not been directly tested previously. Studies on the functions of Sur2 with mouse embryonic fibroblasts (MEFs) showed that there was a multiplicity-dependent growth defect of MAV-1 in Sur2(-/-) MEFs compared to Sur2(+/+) MEFs. Comparison of the viral DNA and viral mRNA levels in Sur2(+/+) and Sur2(-/-) MEFs confirmed that Sur2 was important for efficient viral replication. The viral replication defects in Sur2(-/-) MEFs appeared to be due at least in part to a defect in viral early gene transcription. PMID- 15542642 TI - Improved efficiency of a Salmonella-based vaccine against human papillomavirus type 16 virus-like particles achieved by using a codon-optimized version of L1. AB - Cervical cancer results from cervical infection by human papillomaviruses (HPVs), especially HPV16. An effective vaccine against these HPVs is expected to have a dramatic impact on the incidence of this cancer and its precursor lesions. The leading candidate, a subunit prophylactic HPV virus-like particle (VLP) vaccine, can protect women from HPV infection. An alternative improved vaccine that avoids parenteral injection, that is efficient with a single dose, and that induces mucosal immunity might greatly facilitate vaccine implementation in different settings. In this study, we have constructed a new generation of recombinant Salmonella organisms that assemble HPV16 VLPs and induce high titers of neutralizing antibodies in mice after a single nasal or oral immunization with live bacteria. This was achieved through the expression of a HPV16 L1 capsid gene whose codon usage was optimized to fit with the most frequently used codons in Salmonella. Interestingly, the high immunogenicity of the new recombinant bacteria did not correlate with an increased expression of L1 VLPs but with a greater stability of the L1-expressing plasmid in vitro and in vivo in absence of antibiotic selection. Anti-HPV16 humoral and neutralizing responses were also observed with different Salmonella enterica serovar Typhimurium strains whose attenuating deletions have already been shown to be safe after oral vaccination of humans. Thus, our findings are a promising improvement toward a vaccine strain that could be tested in human volunteers. PMID- 15542644 TI - Epitope determinants of a chimpanzee Fab antibody that efficiently cross neutralizes dengue type 1 and type 2 viruses map to inside and in close proximity to fusion loop of the dengue type 2 virus envelope glycoprotein. AB - The epitope determinants of chimpanzee Fab antibody 1A5, which have been shown to be broadly reactive to flaviviruses and efficient for cross-neutralization of dengue virus type 1 and type 2 (DENV-1 and DENV-2), were studied by analysis of DENV-2 antigenic variants. Sequence analysis showed that one antigenic variant contained a Gly-to-Val substitution at position 106 within the flavivirus conserved fusion peptide loop of the envelope protein (E), and another variant contained a His-to-Gln substitution at position 317 in E. Substitution of Gly(106)Val in DENV-2 E reduced the binding affinity of Fab 1A5 by approximately 80-fold, whereas substitution of His(317)Gln had little or no effect on antibody binding compared to the parental virus. Treatment of DENV-2 with beta mercaptoethanol abolished binding of Fab 1A5, indicating that disulfide bridges were required for the structural integrity of the Fab 1A5 epitope. Binding of Fab 1A5 to DENV-2 was competed by an oligopeptide containing the fusion peptide sequence as shown by competition enzyme-linked immunosorbent assay. Both DENV-2 antigenic variants were shown to be attenuated, or at least similar to the parental virus, when evaluated for growth in cultured cells or for neurovirulence in mice. Fab 1A5 inhibited low pH-induced membrane fusion of mosquito C6/36 cells infected with DENV-1 or DENV-2, as detected by reduced syncytium formation. Both substitutions in DENV-2 E lowered the pH threshold for membrane fusion, as measured in a fusion-from-within assay. In the three-dimensional structure of E, Gly(106) in domain II and His(317) in domain III of the opposite E monomer were spatially close. From the locations of these amino acids, Fab 1A5 appears to recognize a novel epitope that has not been mapped before with a flavivirus monoclonal antibody. PMID- 15542643 TI - Chimpanzee Fab fragments and a derived humanized immunoglobulin G1 antibody that efficiently cross-neutralize dengue type 1 and type 2 viruses. AB - Passive immunization with monoclonal antibodies from humans or nonhuman primates represents an attractive alternative to vaccines for prevention of illness caused by dengue viruses (DENV) and other flaviviruses, including the West Nile virus. In a previous study, repertoire cloning to recover Fab fragments from bone marrow mRNA of chimpanzees infected with all four DENV serotypes (dengue virus serotype 1 [DENV-1] to DENV-4) was described. In that study, a humanized immunoglobulin G1 (IgG1) antibody that efficiently neutralized DENV-4 was recovered and characterized. In this study, the phage library constructed from the chimpanzees was used to recover Fab antibodies against the other three DENV serotypes. Serotype-specific neutralizing Fabs were not identified. Instead, we recovered DENV-neutralizing Fabs that specifically precipitated the envelope protein and were cross-reactive with all four DENV serotypes. Three of the Fabs competed with each other for binding to DENV-1 and DENV-2, although each of these Fabs contained a distinct complementarity determining region 3 (CDR3)-H sequence. Fabs that shared an identical or nearly identical CDR3-H sequences cross-neutralized DENV-1 and DENV-2 at a similar high 50% plaque reduction neutralization test (PRNT(50)) titer, ranging from 0.26 to 1.33 microg/ml, and neutralized DENV-3 and DENV-4 but at a titer 10- to 20-fold lower. One of these Fabs, 1A5, also neutralized the West Nile virus most efficiently among other flaviviruses tested. Fab 1A5 was converted to a full-length antibody in combination with human sequences for production in mammalian CHO cells. Humanized IgG1 1A5 proved to be as efficient as Fab 1A5 for cross-neutralization of DENV-1 and DENV-2 at a titer of 0.48 and 0.95 microg/ml, respectively. IgG1 1A5 also neutralized DENV-3, DENV 4, and the West Nile virus at a PRNT(50) titer of approximately 3.2 to 4.2 microg/ml. This humanized antibody represents an attractive candidate for further development of immunoprophylaxis against DENV and perhaps other flavivirus associated diseases. PMID- 15542645 TI - Comparison of the transcription profile of simian parvovirus with that of the human erythrovirus B19 reveals a number of unique features. AB - Simian parvovirus (SPV) is a member of the genus Erythrovirus and is closely related to the human parvovirus B19. Natural and experimental infection of monkeys with SPV resembles B19 infection of human. We report a detailed characterization of the viral RNAs and proteins generated following transfection of cloned SPV into COS cells and SPV infection of the human erythroid progenitor line UT-7/Epo-S1. SPV and B19 are 50% identical at the nucleotide level, and although their basic transcription and protein expression profiles were generally similar, there were also significant differences. SPV pre-mRNAs contain three introns, compared to two found for B19: an additional intron was found within the capsid-coding region. RNAs in which this intron was spliced were abundant and encoded the SPV 14-kDa protein (analogous to the B19 11-kDa protein), which initiated at an AUG in the exon preceding the third intron. Unlike B19, SPV RNAs were also spliced between the donor of the first intron and the acceptor of the second intron. The third intron was additionally spliced from a portion of these molecules; these mRNAs encoded the 14-kDa protein. A portion was not spliced further and encoded VP2. Like B19, SPV has a polyadenylation signal [AAUAAA (pA)p] in the middle of the genome, which directed efficient polyadenylation of both spliced and unspliced mRNAs (encoding a putative 10-kDa protein, analogous to the B19 7.5-kDa protein, and SPV NS1, respectively). The 14-kDa protein was localized to both in the nucleus and cytoplasm. PMID- 15542646 TI - The DNA architectural protein HMGB1 facilitates RTA-mediated viral gene expression in gamma-2 herpesviruses. AB - Replication and transcription activator (RTA), an immediate-early gene product of gamma-2 herpesviruses including Kaposi's sarcoma-associated herpesvirus (KSHV) and murine gamma herpesvirus 68 (MHV-68), plays a critical role in controlling the viral life cycle. RTA acts as a strong transcription activator for several downstream genes of KSHV and MHV-68 through direct DNA binding, as well as via indirect mechanisms. HMGB1 (also called HMG-1) protein is a highly conserved nonhistone chromatin protein with the ability to bind and bend DNA. HMGB1 protein promoted RTA binding to different RTA target sites in vitro, with greater enhancement to low-affinity sites than to high-affinity sites. Box A or box B and homologues of HMGB1 also enhanced RTA binding to DNA. Transient transfection of HMGB1 stimulated RTA transactivation of RTA-responsive promoters from KSHV and MHV-68. Furthermore, MHV-68 viral gene expression, as well as viral replication, was significantly reduced in HMGB1-deficient cells than in the wild type. This abated viral gene expression was partially restored by HMGB1 transfection into HMGB1(-/-) cells. These results suggest an important function of the DNA architectural protein, HMGB1, in RTA-mediated gene expression, as well as viral replication in gamma-2 herpesviruses. PMID- 15542647 TI - Two modes of pseudorabies virus neuroinvasion and lethality in mice. AB - We describe two distinct modes of neuroinvasion and lethality after murine flank inoculation with virulent and attenuated strains of pseudorabies virus (PRV). Mice infected with virulent (e.g., PRV-Becker, PRV-Kaplan, or PRV-NIA3) strains self-mutilate their flank skin in response to virally induced pruritus, die rapidly with no identifiable symptoms of central nervous system (CNS) infection such as behavioral abnormalities, and have little infectious virus or viral antigen in the brain. In distinct contrast, animals infected with an attenuated PRV vaccine strain (PRV-Bartha) survive approximately three times longer than wild-type PRV-infected animals, exhibit severe CNS abnormalities, and have an abundance of infectious virus in the brain at the time of death. Interestingly, these animals have no skin lesions and do not appear pruritic at any time during infection. The severe pruritus and relatively earlier time until death induced by wild-type PRV infection may reflect the peripheral nervous system (PNS) and immune responses to infection rather than a fatal, virally induced CNS pathology. Based on previously characterized afferent (sensory) and efferent (motor) neuronal pathways that innervate the skin, we deduced that wild-type virulent strains transit through the PNS via both afferent and efferent routes, whereas PRV-Bartha travels by only efferent routes in the PNS en route to the brain. PMID- 15542648 TI - Cyclooxygenase-1 and -2 are required for production of infectious pseudorabies virus. AB - We have recently shown that cyclooxygenase-2 (COX-2) transcription is markedly induced after herpes simplex virus type 1 and pseudorabies virus (PRV) infections of rat embryonic fibroblast (REF) cells (N. Ray and L. W. Enquist, J. Virol. 78:3489-3501, 2004). For this study, we investigated the role of cyclooxygenase induction in the replication and growth of PRV. We demonstrate here a concordant increase in COX-2 mRNA and protein levels after the infection of REF cells. Inhibitors blocking the activity of cyclooxygenases caused a dramatic reduction in PRV growth. Viral growth could be restored if prostaglandin E(2), the final product of COX-2 activity, was added simultaneously with the COX inhibitors. Immediate-early protein IE180, major capsid protein VP5, and glycoprotein expression were slightly reduced in the presence of COX-2 inhibitors, but expression of the early protein EP0 was not affected by COX inhibition. Viral DNA replication was marginally reduced in the presence of a COX-1/2 inhibitor, but there was no defect in viral DNA cleavage. Electron microscopy analysis revealed an increased number of unusual empty capsid structures in the nuclei of cells infected with PRV in the presence of a COX-1/2 inhibitor. These capsid structures shared some characteristics with procapsids but had a novel appearance by negative staining. Our data establish a role for COX-1 and COX-2 in facilitating the efficient growth and replication of PRV in primary cells. PMID- 15542649 TI - Characterization of the outer domain of the gp120 glycoprotein from human immunodeficiency virus type 1. AB - The core of the gp120 glycoprotein from human immunodeficiency virus type 1 (HIV 1) is comprised of three major structural domains: the outer domain, the inner domain, and the bridging sheet. The outer domain is exposed on the HIV-1 envelope glycoprotein trimer and contains binding surfaces for neutralizing antibodies such as 2G12, immunoglobulin G1b12, and anti-V3 antibodies. We expressed the outer domain of HIV-1(YU2) gp120 as an independent protein, termed OD1. OD1 efficiently bound 2G12 and a large number of anti-V3 antibodies, indicating its structural integrity. Immunochemical studies with OD1 indicated that antibody responses against the outer domain of the HIV-1 gp120 envelope glycoprotein are rare in HIV-1-infected human sera that potently neutralize the virus. Surprisingly, such outer-domain-directed antibody responses are commonly elicited by immunization with recombinant monomeric gp120. Immunization with soluble, stabilized HIV-1 envelope glycoprotein trimers elicited antibody responses that more closely resembled those in the sera of HIV-1-infected individuals. These results underscore the qualitatively different humoral immune responses elicited during natural infection and after gp120 vaccination and help to explain the failure of gp120 as an effective vaccine. PMID- 15542650 TI - Interferon regulatory factor 7 is associated with Epstein-Barr virus-transformed central nervous system lymphoma and has oncogenic properties. AB - Interferon regulatory factor 7 (IRF-7) is implicated in the regulation of Epstein Barr virus (EBV) latency. EBV transforms primary B cells, and the major EBV oncoprotein, latent membrane protein 1 (LMP-1), is required for the process. LMP 1 both induces the expression of IRF-7 and activates the IRF-7 protein by phosphorylation and nuclear translocation. Here we report that the expression of IRF-7 is increased in EBV-immortalized B lymphocytes compared with that in primary B cells. IRF-7 was phosphorylated and predominantly localized in the nucleus in the immortalized cells. The expression of IRF-7 was detected in 19 of 27 specimens of primary lymphomas of the human central nervous system by immunohistochemical analysis. The association between LMP-1 and IRF-7 was statistically highly significant for these specimens. An appreciable amount of the IRF-7 expressed in lymphoma cells was localized in the nucleus. Furthermore, IRF-7 promoted the anchorage-independent growth of NIH 3T3 cells. LMP-1 and IRF-7 showed additive effects on the growth transformation of NIH 3T3 cells. IRF-7 expressing NIH 3T3 cells formed tumors in athymic mice. Thus, IRF-7 has oncogenic properties and, along with LMP-1, may mediate or potentiate the EBV transformation process in the pathogenesis of EBV-associated lymphomas. PMID- 15542651 TI - Inhibition of human immunodeficiency virus replication by a dual CCR5/CXCR4 antagonist. AB - Here we report that the N-pyridinylmethyl cyclam analog AMD3451 has antiviral activity against a wide variety of R5, R5/X4, and X4 strains of human immunodeficiency virus type 1 (HIV-1) and HIV-2 (50% inhibitory concentration [IC(50)] ranging from 1.2 to 26.5 microM) in various T-cell lines, CCR5- or CXCR4 transfected cells, peripheral blood mononuclear cells (PBMCs), and monocytes/macrophages. AMD3451 also inhibited R5, R5/X4, and X4 HIV-1 primary clinical isolates in PBMCs (IC(50), 1.8 to 7.3 microM). A PCR-based viral entry assay revealed that AMD3451 blocks R5 and X4 HIV-1 infection at the virus entry stage. AMD3451 dose-dependently inhibited the intracellular Ca(2+) signaling induced by the CXCR4 ligand CXCL12 in T-lymphocytic cells and in CXCR4 transfected cells, as well as the Ca(2+) flux induced by the CCR5 ligands CCL5, CCL3, and CCL4 in CCR5-transfected cells. The compound did not interfere with chemokine-induced Ca(2+) signaling through CCR1, CCR2, CCR3, CCR4, CCR6, CCR9, or CXCR3 and did not induce intracellular Ca(2+) signaling by itself at concentrations up to 400 microM. In freshly isolated monocytes, AMD3451 inhibited the Ca(2+) flux induced by CXCL12 and CCL4 but not that induced by CCL2, CCL3, CCL5, and CCL7. The CXCL12- and CCL3-induced chemotaxis was also dose-dependently inhibited by AMD3451. Furthermore, AMD3451 inhibited CXCL12- and CCL3L1-induced endocytosis in CXCR4- and CCR5-transfected cells. AMD3451, in contrast to the specific CXCR4 antagonist AMD3100, did not inhibit but enhanced the binding of several anti-CXCR4 monoclonal antibodies (such as clone 12G5) at the cell surface, pointing to a different interaction with CXCR4. AMD3451 is the first low molecular-weight anti-HIV agent with selective HIV coreceptor, CCR5 and CXCR4, interaction. PMID- 15542652 TI - Differential rescue of poliovirus RNA replication functions by genetically modified RNA polymerase precursors. AB - We have previously described the RNA replication properties of poliovirus transcripts harboring chimeric RNA polymerase sequences representing suballelic exchanges between poliovirus type 1 (PV1) and coxsackievirus B3 (CVB3) utilizing an in vitro translation and RNA replication assay (C. Cornell, R. Perera, J. E. Brunner, and B. L. Semler, J. Virol. 78:4397-4407, 2004). We showed that three of the seven chimeras were capable of RNA replication in vitro, although replication levels were greatly reduced compared to that of wild-type transcripts. Interestingly, one of the replication-competent transcripts displayed a strand specific RNA synthesis defect suggesting (i) a differential replication complex assembly mechanism involving 3D and/or precursor molecules (i.e., 3CD) required for negative- versus positive-strand RNA synthesis or (ii) effect(s) on the ability of the 3D polymerase to form higher-ordered structures required for positive-strand RNA synthesis. In this study, we have attempted to rescue defective RNA replication in vitro by cotranslating nonstructural proteins from a transcript encoding a large precursor polyprotein (P3) to complement 3D polymerase and/or precursor polypeptide functions altered in each of the chimeric constructs. Utilization of a wild-type P3 construct revealed that all transcripts containing chimeric PV1/CVB3 polymerase sequences can be complemented in trans for both negative- and positive-strand RNA synthesis. Furthermore, data from experiments utilizing genetically modified forms of the P3 polyprotein, containing mutations within 3C or 3D sequences, strongly suggest the existence of different protein-protein and protein-RNA interactions required for positive- versus negative-strand RNA synthesis. These results, combined with data from in vitro RNA elongation assays, indicate that the delivery of active 3D RNA polymerase to replication complexes requires a series of macromolecular interactions that rely on the presence of specific 3D amino acid sequences. PMID- 15542653 TI - Structural protein requirements in equine arteritis virus assembly. AB - Equine arteritis virus (EAV) is an enveloped, positive-stranded RNA virus belonging to the family Arteriviridae of the order Nidovirales. EAV particles contain seven structural proteins: the nucleocapsid protein N, the unglycosylated envelope proteins M and E, and the N-glycosylated membrane proteins GP(2b) (previously named G(S)), GP(3), GP(4), and GP(5) (previously named G(L)). Proteins N, M, and GP(5) are major virion components, E occurs in virus particles in intermediate amounts, and GP(4), GP(3), and GP(2b) are minor structural proteins. The M and GP(5) proteins occur in virus particles as disulfide-linked heterodimers while the GP(4), GP(3), and GP(2b) proteins are incorporated into virions as a heterotrimeric complex. Here, we studied the effect on virus assembly of inactivating the structural protein genes one by one in the context of a (full-length) EAV cDNA clone. It appeared that the three major structural proteins are essential for particle formation, while the other four virion proteins are dispensable. When one of the GP(2b), GP(3), or GP(4) proteins was missing, the incorporation of the remaining two minor envelope glycoproteins was completely blocked while that of the E protein was greatly reduced. The absence of E entirely prevented the incorporation of the GP(2b), GP(3), and GP(4) proteins into viral particles. EAV particles lacking GP(2b), GP(3), GP(4), and E did not markedly differ from wild-type virions in buoyant density, major structural protein composition, electron microscopic appearance, and genomic RNA content. On the basis of these results, we propose a model for the EAV particle in which the GP(2b)/GP(3)/GP(4) heterotrimers are positioned, in association with a defined number of E molecules, above the vertices of the putatively icosahedral nucleocapsid. PMID- 15542654 TI - Induction of Epstein-Barr virus latent membrane protein 1 by a lytic transactivator Rta. AB - Latent membrane protein 1 (LMP1) of Epstein-Barr virus (EBV) is a transforming protein that affects multiple cell signaling pathways and contributes to EBV associated oncogenesis. LMP1 can be expressed in some states of EBV latency, and significant induction of full-length LMP1 is also observed frequently during virus reactivation into the lytic cycle. It is still unknown how LMP1 expression is regulated during the lytic stage and whether any EBV lytic protein is involved in the induction of LMP1. In this study, we first identified that LMP1 expression is associated with the spontaneous virus reactivation in EBV-infected 293 cells and that its expression is a downstream event of the lytic cycle. We further found that LMP1 can be induced by ectopic expression of Rta, an EBV immediate early lytic protein. The Rta-mediated LMP1 induction is independent of another immediate-early protein, Zta. Northern blotting and reverse transcription-PCR analysis revealed that Rta upregulates LMP1 at the RNA level. Reporter gene assays further demonstrated that Rta activates both the proximal and distal promoters of the LMP1 gene in EBV-negative cells. Both the amino and carboxyl termini of the Rta protein are required for the induction of LMP1. In addition, Rta transactivates LMP1 not only in epithelial cells but also in B-lymphoid cells. This study reveals a new mechanism to upregulate LMP1 expression, expanding the knowledge of LMP1 regulation in the EBV life cycle. Considering an equivalent case of Kaposi's sarcoma-associated herpesvirus, induction of a transforming membrane protein by a key lytic transactivator during virus reactivation is likely to be a conserved event for gammaherpesviruses. PMID- 15542655 TI - Immunogenicity and protection efficacy of replication-deficient influenza A viruses with altered NS1 genes. AB - We explored the immunogenic properties of influenza A viruses with altered NS1 genes (NS1 mutant viruses). NS1 mutant viruses expressing NS1 proteins with an impaired RNA-binding function or insertion of a longer foreign sequence did not replicate in murine lungs but still were capable of inducing a Th1-type immune response resulting in significant titers of virus-specific serum and mucosal immunoglobulin G2 (IgG2) and IgA, but with lower titers of IgG1. In contrast, replicating viruses elicited high titers of serum and mucosal IgG1 but less serum IgA. Replication-deficient NS1 mutant viruses induced a rapid local release of proinflammatory cytokines such as interleukin-1beta (IL-1beta) and IL-6. Moreover, these viruses also elicited markedly higher levels of IFN-alpha/beta in serum than the wild-type virus. Comparable numbers of virus-specific primary CD8(+) T cells were determined in all of the groups of immunized mice. The most rapid onset of the recall CD8(+)-T-cell response upon the wild-type virus challenge was detected in mice primed with NS1 mutant viruses eliciting high levels of cytokines. It is noteworthy that there was one NS1 mutant virus encoding NS1 protein with a deletion of 40 amino acids predominantly in the RNA binding domain that induced the highest levels of IFN-alpha/beta, IL-6 and IL 1beta after infection. Mice that were immunized with this virus were completely protected from the challenge infection. These findings indicate that a targeted modification of the RNA-binding domain of the NS1 protein is a valuable technique to generate replication-deficient, but immunogenic influenza virus vaccines. PMID- 15542656 TI - Cell-type-dependent effect of transforming growth factor beta, a major cytokine in breast milk, on human immunodeficiency virus type 1 infection of mammary epithelial MCF-7 cells or macrophages. AB - Breastfeeding plays a substantial role in mother-to-child transmission of human immunodeficiency virus type 1 (HIV-1). Mammary epithelial cells, as well as macrophages and lymphocytes, are thought to serve as sources of the virus in breast milk. Soluble factors in breast milk exert various biological functions, including immune tolerance or immune modulation, and may influence milk-borne infection with HIV-1. In this study we show that transforming growth factor beta (TGF-beta), a major cytokine in breast milk, inhibited HIV-1 infection of mammary epithelial MCF-7 cells but enhanced that of macrophages. TGF-beta downregulated the HIV-1 long terminal repeat (LTR) promoter in MCF-7 cells but upregulated it in macrophages. Stimulation with TGF-beta suppressed NF-kappaB binding to the HIV 1 LTR in MCF-7 cells, at least in part by downregulating induced IkappaB kinase expression. Cell type-dependent effects of TGF-beta on HIV-1 expression may play a role in milk-borne infection with HIV-1. PMID- 15542657 TI - Amino acid substitutions in the F-specific domain in the stalk of the newcastle disease virus HN protein modulate fusion and interfere with its interaction with the F protein. AB - The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus mediates attachment to sialic acid receptors, as well as cleavage of the same moiety. HN also interacts with the other viral glycoprotein, the fusion (F) protein, to promote membrane fusion. The ectodomain of the HN spike consists of a stalk and a terminal globular head. The most conserved part of the stalk consists of two heptad repeats separated by a nonhelical intervening region (residues 89 to 95). Several amino acid substitutions for a completely conserved proline residue in this region not only impair fusion and the HN-F interaction but also decrease neuraminidase activity in the globular domain, suggesting that the substitutions may alter HN structure. Substitutions for L94 also interfere with fusion and the HN-F interaction but have no significant effect on any other HN function. Amino acid substitutions at other positions in the intervening region also modulate only fusion. In all cases, diminished fusion correlates with a decreased ability of the mutated HN protein to interact with F at the cell surface. These findings indicate that the intervening region is critical to the role of HN in the promotion of fusion and may be directly involved in its interaction with the homologous F protein. PMID- 15542658 TI - Identification of a mouse cytomegalovirus gene selectively targeting CD86 expression on antigen-presenting cells. AB - We and others have shown that infection of dendritic cells with murine cytomegalovirus (MCMV) leads to severe functional impairment of these antigen presenting cells (D. M. Andrews, C. E. Andoniou, F. Granucci, P. Ricciardi Castagnoli, and M. A. Degli-Esposti, Nat. Immunol. 2:1077-1084, 2001; S. Mathys, T. Schroeder, J. Ellwart, U. H. Koszinowski, M. Messerle, and U. Just, J. Infect. Dis. 187:988-999, 2003). Phenotypically, reduced surface expression of costimulatory molecules and major histocompatibility complex molecules was detected. In order to identify the molecular basis for the observed effects, we generated MCMV mutants with large deletions of nonessential genes. The study was facilitated by the finding that a monocyte-macrophage cell line displayed similar phenotypic alterations after MCMV infection. By analyzing the expression of cell surface molecules on infected cells, we identified a mutant virus which is no longer able to downmodulate the expression of the costimulatory molecule CD86. Additional mutants with smaller deletions allowed us to pin down the responsible gene to a certain genomic region. RNA analysis led to the identification of the spliced gene m147.5, encoding a protein with 145 amino acids. Experiments with an m147.5 mutant revealed that the protein affects CD86 expression only, suggesting that additional MCMV genes are responsible for downmodulation of the other surface molecules. Identification of viral gene products interfering with functionally important proteins of antigen-presenting cells will provide the basis to dissect the complex interaction of CMV with these important cells and to evaluate the biological importance of these viral genes in vivo. PMID- 15542659 TI - Lentiviral vectors interfering with virus-induced CD4 down-modulation potently block human immunodeficiency virus type 1 replication in primary lymphocytes. AB - CD4 down-modulation is essential for the production of human immunodeficiency virus (HIV) infectious particles. Disease progression correlates with enhanced viral induced CD4 down-modulation, and a subset of long-term nonprogressors carry viruses defective in this function. Despite multiple pieces of evidence highlighting the importance of this function in viral pathogenesis in vivo, to date, HIV-induced CD4 down-modulation has not been used as a target for intervention. We describe here HIV-based vectors that deliver truncated CD4 molecules resistant to down-modulation by the viral products Nef and Vpu. Infection of cells previously transduced with these vectors proceeded normally, and viral particles were released in normal amounts. However, the infectivity of the released virions was reduced 1,000-fold. Lentiviral vectors expressing truncated CD4 molecules were efficient at blocking HIV-1 infectivity and replication in several cell lines and in CD4-positive primary lymphocytes. The findings presented here provide proof-of-principle that approaches targeting the virus-induced CD4 down-modulation may constitute the basis for novel anti-HIV therapies. PMID- 15542660 TI - CD4+ CD25+ T cells regulate vaccine-generated primary and memory CD8+ T-cell responses against herpes simplex virus type 1. AB - It has become evident that naturally occurring CD25(+) regulatory T cells (T(reg) cells) not only influence self-antigen specific immune response but also dampen foreign antigen specific immunity. This report extends our previous findings by demonstrating that immunity to certain herpes simplex virus (HSV) vaccines is significantly elevated and more effective if T(reg) cell response is curtailed during either primary or recall immunization. The data presented here show that removal of CD25(+) T(reg) cells prior to SSIEFARL-CpG or gB-DNA immunization significantly enhanced the resultant CD8(+) T-cell response to the immunodominant SSIEFARL peptide. The enhanced CD8(+) T-cell reactivity in T(reg) cell-depleted animals was between two- and threefold and evident in both acute and memory stages. Interestingly, removal of CD25(+) T(reg) cells during the memory recall response to plasmid immunization resulted in a twofold increase in CD8(+) T-cell memory pool. Moreover, in the challenge experiments, memory CD8(+) T cells generated with plasmid DNA in the absence of T(reg) cells cleared the virus more effectively compared with control groups. We conclude that CD25(+) T(reg) cells quantitatively as well as qualitatively affect the memory CD8(+) T-cell response generated by gB-DNA vaccination against HSV. However, it remains to be seen if all types of vaccines against HSV are similarly affected by CD25(+) T(reg) cells and if it is possible to devise means of limiting T(reg) cell activity to enhance vaccine efficacy. PMID- 15542661 TI - Bracoviruses contain a large multigene family coding for protein tyrosine phosphatases. AB - The relationship between parasitic wasps and bracoviruses constitutes one of the few known mutualisms between viruses and eukaryotes. The virions produced in the wasp ovaries are injected into host lepidopteran larvae, where virus genes are expressed, allowing successful development of the parasite by inducing host immune suppression and developmental arrest. Bracovirus-bearing wasps have a common phylogenetic origin, and contemporary bracoviruses are hypothesized to have been inherited by chromosomal transmission from a virus that originally integrated into the genome of the common ancestor wasp living 73.7 +/- 10 million years ago. However, so far no conserved genes have been described among different braconid wasp subfamilies. Here we show that a gene family is present in bracoviruses of different braconid wasp subfamilies (Cotesia congregata, Microgastrinae, and Toxoneuron nigriceps, Cardiochilinae) which likely corresponds to an ancient component of the bracovirus genome that might have been present in the ancestral virus. The genes encode proteins belonging to the protein tyrosine phosphatase family, known to play a key role in the control of signal transduction pathways. Bracovirus protein tyrosine phosphatase genes were shown to be expressed in different tissues of parasitized hosts, and two protein tyrosine phosphatases were produced with recombinant baculoviruses and tested for their biochemical activity. One protein tyrosine phosphatase is a functional phosphatase. These results strengthen the hypothesis that protein tyrosine phosphatases are involved in virally induced alterations of host physiology during parasitism. PMID- 15542662 TI - Separate sequences in a murine retroviral envelope protein mediate neuropathogenesis by complementary mechanisms with differing requirements for tumor necrosis factor alpha. AB - The innate immune response, through the induction of proinflammatory cytokines and antiviral factors, plays an important role in protecting the host from pathogens. Several components of the innate response, including tumor necrosis factor alpha (TNF-alpha), monocyte chemoattractant protein 1, interferon inducible protein 10, and RANTES, are upregulated in the brain following neurovirulent retrovirus infection in humans and in animal models. However, it remains unclear whether this immune response is protective, pathogenic, or both. In the present study, by using TNF-alpha(-/-) mice we analyzed the contribution of TNF-alpha to neurological disease induced by four neurovirulent murine retroviruses, with three of these viruses encoding portions of the same neurovirulent envelope protein. Surprisingly, only one retrovirus (EC) required TNF-alpha for disease induction, and this virus induced less TNF-alpha expression in the brain than did the other retroviruses. Analysis of glial fibrillary acidic protein and F4/80 in EC-infected TNF-alpha(-/-) mice showed normal activation of astrocytes but not of microglia. Thus, TNF-alpha-mediated microglial activation may be important in the pathogenic process initiated by EC infection. In contrast, TNF-alpha was not required for pathogenesis of the closely related BE virus and the BE virus induced disease in TNF-alpha(-/-) mice by a different mechanism that did not require microglial activation. These results provide new insights into the multifactorial mechanisms involved in retrovirus-induced neurodegeneration and may also have analogies to other types of neurodegeneration. PMID- 15542663 TI - Inhibition of tumor necrosis factor (TNF) signal transduction by the adenovirus group C RID complex involves downregulation of surface levels of TNF receptor 1. AB - Adenoviruses employ multiple genes to inhibit the host antiviral responses. There is increasing evidence that these immunoregulatory genes may function either during lytic or latent infection. Adenovirus early transcription region 3 (E3) encodes at least seven proteins, five of which block the acquired or innate immune response. Previous findings from this laboratory demonstrated that the E3 proteins 10.4K and 14.5K, which form a complex in the plasma membrane, inhibit tumor necrosis factor (TNF)-induced activation of NF-kappaB and the synthesis of chemokines. To determine the mechanism of inhibition of these pathways by the adenovirus E3 10.4K/14.5K proteins, we have examined the effects of this viral complex on the inhibition of AP-1 and NF-kappaB activation by TNF and found a reduction in assembly of the TNF receptor 1 (TNFR1) signaling complex at the plasma membrane accompanied by downregulation of surface levels of TNFR1. PMID- 15542664 TI - Requirement of heat shock protein 90 for human hepatitis B virus reverse transcriptase function. AB - The initiation of reverse transcription and nucleocapsid assembly in hepatitis B virus (HBV) depends on the specific recognition of an RNA signal (the packaging signal, epsilon) on the pregenomic RNA (pgRNA) by the viral reverse transcriptase (RT). RT-epsilon interaction in the duck hepatitis B virus (DHBV) was recently shown to require the molecular chaperone complex, the heat shock protein 90 (Hsp90). However, the requirement for RT-epsilon interaction in the human HBV has remained unknown due to the inability to obtain a purified RT protein active in specific epsilon binding. We now report that Hsp90 is also required for HBV RT epsilon interaction. Inhibition of Hsp90 led to diminished HBV pgRNA packaging into nucleocapsids in cells, which depends on RT-epsilon interaction. Furthermore, using truncated HBV RT proteins purified from bacteria and five purified Hsp90 chaperone factors, we have developed an in vitro RT-epsilon binding assay. Our results demonstrate that Hsp90, in a dynamic process that was dependent on ATP hydrolysis, facilitated RT-epsilon interaction in HBV, as in DHBV. Specific epsilon binding required sequences from both the amino-terminal terminal protein and the carboxy-terminal RT domain. Only the cognate HBV epsilon, but not the DHBV epsilon, could bind the HBV RT proteins. Furthermore, the internal bulge, but not the apical loop, of epsilon was required for RT binding. The establishment of a defined in vitro reconstitution system has now paved the way for future biochemical and structural studies to elucidate the mechanisms of RT-epsilon interaction and chaperone activation. PMID- 15542665 TI - JNK regulates the release of proapoptotic mitochondrial factors in reovirus infected cells. AB - Reovirus-induced apoptosis is associated with activation of the proapoptotic mitogen-activated protein kinase c-Jun N-terminal kinase (JNK) and the JNK associated transcription factor c-Jun. Here we show that reovirus-induced apoptosis and activation of caspase 3 are inhibited in cells deficient in MEK kinase 1, an upstream activator of JNK in reovirus-infected cells. Inhibition of JNK activity following reovirus infection delays the release of proapoptotic mitochondrial factors and the subsequent onset of apoptosis. In contrast, reovirus-induced apoptosis is not blocked by infection with adenovirus expressing dominant-negative c-Jun, and c-Jun activation does not correlate with apoptosis in reovirus-infected cells. This is the first report demonstrating that JNK is associated with regulation of mitochondrial pathways of apoptosis following viral infection. PMID- 15542666 TI - Spread and replication of and immune response to gamma134.5-negative herpes simplex virus type 1 vectors in BALB/c mice. AB - We have previously shown that intracranial infection of herpes simplex virus type 1 (HSV-1) vector R8306 expressing interleukin-4 (IL-4) can abolish symptoms of experimental autoimmune encephalomyelitis, which is used as a model for human multiple sclerosis (Broberg et al., Gene Ther. 8:769-777, 2001). The aim of the current study was to search for means other than intracranial injection to deliver HSV-derived vectors to the central nervous system of mice. We also aimed to study the replication efficiency of these vectors in nervous system tissues and to elucidate the effects of the viruses on the immune response. We studied the spread and replication of the following viruses with deletions in neurovirulence gene gamma(1)34.5: R3616, R849 (lacZ transgene), R3659 (alpha-tk), R8306 (murine IL-4 transgene), and R8308 (murine IL-10 transgene). The samples were taken from trigeminal ganglia and brains of BALB/c mice after corneal, intralabial, and intranasal infection, and the viral load was examined by viral culture, HSV DNA PCR, and VP16 reverse transcription (RT)-PCR. The results show that (i) intranasal infection was the most efficient means of spread to the central nervous system (CNS) besides intracranial injection; (ii) the viruses did not grow in the culture from the brain samples, but the viral DNA persisted even until day 21 postinfection; (iii) viral replication, as observed by VP16 mRNA RT PCR, occurred mainly on days 4 and 7 postinfection in trigeminal ganglia and to a low extent in brain; (iv) R3659, R8306, and R8308 showed reactivation from the trigeminal ganglia in explant cultures; (v) in the brain, the vectors spread to the midbrain more efficiently than to other brain areas; and (vi) the deletions in the R3659 genome significantly limited the ability of this virus to replicate in the nervous system. The immunological studies show that (i) the only recombinant to induce IL-4 mRNA expression in the brain was R8306, the gamma interferon response was very low in the brain for R3659 and R8306, and the IL 23p19 response to R8306 decreased by day 21 postinfection, unlike for the other viruses; (ii) Deltagamma(1)34.5 HSV vectors modulated the subsets of the splenocytes differently depending on the transgene; (iii) R3659 infection of the nervous system induces expression and production of cytokines from the stimulated splenocytes; and (iv) HSV vectors expressing IL-4 or IL-10 induce expression and production of both of the Th2-type cytokines from splenocytes. We conclude that the intranasal route of infection is a possible means of delivery of Deltagamma(1)34.5 HSV vectors to the CNS in addition to intracranial infection, although replication in the CNS remains minimal. The DNA of the HSV vectors is able to reside in the brain for at least 3 weeks. The features of the immune response to the vectors must be considered and may be exploited in gene therapy experiments with these vectors. PMID- 15542667 TI - SYNCRIP, a member of the heterogeneous nuclear ribonucleoprotein family, is involved in mouse hepatitis virus RNA synthesis. AB - Several cellular proteins, including several heterogeneous nuclear ribonucleoproteins (hnRNPs), have been shown to function as regulatory factors for mouse hepatitis virus (MHV) RNA synthesis as a result of their binding to the 5' and 3' untranslated regions (UTRs) of the viral RNA. Here, we identified another cellular protein, p70, which has been shown by UV cross-linking to bind both the positive- and negative-strand UTRs of MHV RNA specifically. We purified p70 with a a one-step RNA affinity purification procedure with the biotin-labeled 5'-UTR. Matrix-assisted laser desorption ionization (MALDI)-mass spectrometry identified it as synaptotagmin-binding cytoplasmic RNA-interacting protein (SYNCRIP). SYNCRIP is a member of the hnRNP family and localizes largely in the cytoplasm. The p70 was cross-linked to the MHV positive- or negative-strand UTR in vitro and in vivo. The bacterially expressed SYNCRIP was also able to bind to the 5'-UTR of both strands. The SYNCRIP-binding site was mapped to the leader sequence of the 5'-UTR, requiring the UCUAA repeat sequence. To investigate the functional significance of SYNCRIP in MHV replication, we expressed a full-length or a C-terminally truncated form of SYNCRIP in mammalian cells expressing the MHV receptor. The overexpression of either form of SYNCRIP inhibited syncytium formation induced by MHV infection. Furthermore, downregulation of the endogenous SYNCRIP with a specific short interfering RNA delayed MHV RNA synthesis; in contrast, overexpression or downregulation of SYNCRIP did not affect MHV translation. These results suggest that SYNCRIP may be directly involved in MHV RNA replication as a positive regulator. This study identified an additional cellular hnRNP as an MHV RNA-binding protein potentially involved in viral RNA synthesis. PMID- 15542668 TI - Gender differences in human immunodeficiency virus type 1-specific CD8 responses in the reproductive tract and colon following nasal peptide priming and modified vaccinia virus Ankara boosting. AB - Induction of mucosal anti-human immunodeficiency virus type 1 (HIV-1) T-cell responses in males and females will be important for the development of a successful HIV-1 vaccine. An HIV-1 envelope peptide, DNA plasmid, and recombinant modified vaccinia virus Ankara (rMVA) expressing the H-2D(d)-restricted cytotoxic T lymphocyte P18 epitope were used as immunogens to test for their ability to prime and boost anti-HIV-1 T-cell responses at mucosal and systemic sites in BALB/c mice. We found of all prime-boost combinations tested, an HIV-1 Env peptide subunit mucosal prime followed by systemic (intradermal) boosting with rMVA yielded the maximal induction of gamma interferon (IFN-gamma) spot-forming cells in the female genital tract and colon. However, this mucosal prime-systemic rMVA boost regimen was minimally immunogenic for the induction of genital, colon, or lung anti-HIV-1 T-cell responses in male mice. We determined that a mucosal Env subunit immunization could optimally prime an rMVA boost in female but not male mice, as determined by the magnitude of antigen-specific IFN-gamma responses in the reproductive tracts, colon, and lung. Defective mucosal priming in male mice could not be overcome by multiple mucosal immunizations. However, rMVA priming followed by an rMVA boost was the optimal prime-boost strategy for male mice as determined by the magnitude of antigen-specific IFN-gamma responses in the reproductive tract and lung. Thus, prime-boost immunization strategies able to induce mucosal antigen-specific IFN-gamma responses were identified for male and female mice. Understanding the cellular and molecular basis of gender determined immune responses will be important for optimizing induction of anti HIV-1 mucosal immune responses in both males and females. PMID- 15542669 TI - Human cytomegalovirus-specific CD4+-T-cell cytokine response induces fractalkine in endothelial cells. AB - Cytomegalovirus (CMV) infection has been linked to inflammation-related disease processes in the human host, including vascular diseases and chronic transplant rejection. The mechanisms through which CMV affects the pathogenesis of these diseases are for the most part unknown. To study the contributing role of the host immune response to CMV in these chronic inflammatory processes, we examined endothelial cell interactions with peripheral blood mononuclear cells (PBMC). Endothelial cultures were monitored for levels of fractalkine induction as a marker for initiating the host inflammatory response. Our results demonstrate that in the presence of CMV antigen PBMC from normal healthy CMV-seropositive donors produce soluble factors that induce fractalkine in endothelial cells. This was not observed in parallel assays with PBMC from seronegative donors. Examination of subset populations within the PBMC further revealed that CMV antigen-stimulated CD4(+) T cells were the source of the factors, gamma interferon and tumor necrosis factor alpha, driving fractalkine induction. Direct contact between CD4(+) cells and the endothelial monolayers is required for this fractalkine induction, where the endothelial cells appear to provide antigen presentation functions. These findings indicate that CMV may represent one member of a class of persistent pathogens where the antigen-specific T-cell response can result in the induction of fractalkine, leading to chronic inflammation and endothelial cell injury. PMID- 15542670 TI - Human cytomegalovirus 5-kilobase immediate-early RNA is a stable intron. AB - Immediate-early viral gene products of human cytomegalovirus (HCMV) are derived from several genomic loci and largely serve to establish a cellular environment conducive to viral replication. We have further examined an unusual immediate early transcript known as the 5-kb RNA, concluding that it is a stable intron encoded by HCMV. The 5-kb RNA is highly AT rich in sequence and lacks open reading frames likely to be translated into protein. We confirmed the absence of polyadenylation of the transcript and showed that it is primarily nuclear localized during viral infection. We mapped the 5' end of the 5-kb RNA to a consensus splice donor site and localized the 3' end in the vicinity of a splice acceptor site. In transfection studies, we showed that the 5-kb RNA can be spliced from a heterologous primary transcript. Using bacterial artificial chromosome technology, we constructed a viral recombinant containing a mutation in the 5' splice donor site that defines the 5' end of the RNA and found that this mutation eliminates expression of the 5-kb RNA during viral infection. This mutant grows in human fibroblasts without complementation. Taken together, these data support the conclusion that the 5-kb RNA is a stable intron expressed by HCMV. PMID- 15542671 TI - Sequence variation within the dominant amino terminus epitope affects antibody binding and neutralization of human immunodeficiency virus type 1 Tat protein. AB - Tat is among the required regulatory genes of human immunodeficiency virus type 1 (HIV-1). Tat functions both within infected cells as a transcription factor and as an extracellular factor that binds and alters bystander cells. Some functions of extracellular Tat can be neutralized by immune serum or monoclonal antibodies. In order to understand the antibody response to Tat, we are defining antibody epitopes and the effects of natural Tat sequence variation on antibody recognition. The dominant Tat epitope in macaque sera is within the first 15 amino acids of the protein amino terminus. Together with a subdominant response to amino acids 57 to 60, these two regions account for most of the macaque response to linear Tat epitopes and both regions are also sites for the binding of neutralizing antibodies. However, the dominant and subdominant epitope sequences differ among virus strains, and this natural variation can preclude antibody binding and Tat neutralization. We also examined serum samples from 31 HIV-positive individuals that contained Tat binding antibodies; 23 of the 31 sera recognized the amino terminus peptide. Similar to binding in macaques, human antibody binding to the amino terminus was affected by variations at positions 7 and 12, sequences that are distinct for clade B compared to other viral clades. Tat-neutralizing antibodies to the dominant amino terminus epitope are affected by HIV clade variation. PMID- 15542672 TI - Expression and characterization of a soluble form of tomato spotted wilt virus glycoprotein GN. AB - Tomato spotted wilt virus (TSWV), a member of the Tospovirus genus within the Bunyaviridae, is an economically important plant pathogen with a worldwide distribution. TSWV is transmitted to plants via thrips (Thysanoptera: Thripidae), which transmit the virus in a persistent propagative manner. The envelope glycoproteins, G(N) and G(C), are critical for the infection of thrips, but they are not required for the initial infection of plants. Thus, it is assumed that the envelope glycoproteins play important roles in the entry of TSWV into the insect midgut, the first site of infection. To directly test the hypothesis that G(N) plays a role in TSWV acquisition by thrips, we expressed and purified a soluble, recombinant form of the G(N) protein (G(N)-S). The expression of G(N)-S allowed us to examine the function of G(N) in the absence of other viral proteins. We detected specific binding to thrips midguts when purified G(N)-S was fed to thrips in an in vivo binding assay. The TSWV nucleocapsid protein and human cytomegalovirus glycoprotein B did not bind to thrips midguts, indicating that the G(N)-S-thrips midgut interaction is specific. TSWV acquisition inhibition assays revealed that thrips that were concomitantly fed purified TSWV and G(N)-S had reduced amounts of virus in their midguts compared to thrips that were fed TSWV only. Our findings that G(N)-S binds to larval thrips guts and decreases TSWV acquisition provide evidence that G(N) may serve as a viral ligand that mediates the attachment of TSWV to receptors displayed on the epithelial cells of the thrips midgut. PMID- 15542673 TI - Novel replication-incompetent vector derived from adenovirus type 11 (Ad11) for vaccination and gene therapy: low seroprevalence and non-cross-reactivity with Ad5. AB - A novel plasmid-based adenovirus vector system that enables manufacturing of replication-incompetent (DeltaE1) adenovirus type 11 (Ad11)-based vectors is described. Ad11 vectors are produced on PER.C6/55K cells yielding high-titer vector batches after purification. Ad11 seroprevalence proves to be significantly lower than that of Ad5, and neutralizing antibody titers against Ad11 are low. Ad11 seroprevalence among human immunodeficiency virus-positive (HIV(+)) individuals is as low as that among HIV(-) individuals, independent of the level of immune suppression. The low level of coinciding seroprevalence between Ad11 and Ad35 in addition to a lack of correlation between high neutralizing antibody titers towards either adenovirus strongly suggest that the limited humoral cross reactive immunity between these two highly related B viruses appears not to preclude the use of both vectors in the same individual. Ad11 transduces primary cells including smooth muscle cells, synoviocytes, and dendritic cells and cardiovascular tissues with higher efficiency than Ad5. Ad11 and Ad35 appear to have a similar tropism as judged by green fluorescent protein expression levels determined by using a panel of cancer cell lines. In addition, Ad5 preimmunization did not significantly affect Ad11-mediated transduction in C57BL/6 mice. We therefore conclude that the Ad11-based vector represents a novel and useful candidate gene transfer vehicle for vaccination and gene therapy. PMID- 15542674 TI - Mutation of all Runx (AML1/core) sites in the enhancer of T-lymphomagenic SL3-3 murine leukemia virus unmasks a significant potential for myeloid leukemia induction and favors enhancer evolution toward induction of other disease patterns. AB - SL3-3 murine leukemia virus is a potent inducer of T-lymphomas in mice. Using inbred NMRI mice, it was previously reported that a mutant of SL3-3 with all enhancer Runx (AML1/core) sites disrupted by 3-bp mutations (SL3-3dm) induces predominantly non-T-cell tumors with severely extended latency (S. Ethelberg, J. Lovmand, J. Schmidt, A. Luz, and F. S. Pedersen, J. Virol. 71:7273-7280, 1997). By use of three-color flow cytometry and molecular and histopathological analyses, we have now performed a detailed phenotypic characterization of SL3-3- and SL3-3dm-induced tumors in this mouse strain. All wild-type induced tumors had clonal T-cell receptor beta rearrangements, and the vast majority were CD3(+) CD4(+) CD8(-) T-lymphomas. Such a consistent phenotypic pattern is unusual for murine leukemia virus-induced T-lymphomas. The mutant virus induced malignancies of four distinct hematopoietic lineages: myeloid, T lymphoid, B lymphoid, and erythroid. The most common disease was myeloid leukemia with maturation. Thus, mutation of all Runx motifs in the enhancer of SL3-3 severely impedes viral T lymphomagenicity and thereby discloses a considerable and formerly unappreciated potential of this virus for myeloid leukemia induction. Proviral enhancers with complex structural alterations (deletions, insertions, and/or duplications) were found in most SL3-3dm-induced T-lymphoid tumors and immature myeloid leukemias but not in any cases of myeloid leukemia with maturation, mature B-lymphoma, or erythroleukemia. Altogether, our results indicate that the SL3-3dm enhancer in itself promotes induction of myeloid leukemia with maturation but that structural changes may arise in vivo and redirect viral disease specificity to induction of T-lymphoid or immature myeloid leukemias, which typically develop with moderately shorter latencies. PMID- 15542676 TI - Comparative analysis of signal transduction by CD40 and the Epstein-Barr virus oncoprotein LMP1 in vivo. AB - There is much evidence, based primarily on in vitro studies, indicating that the Epstein-Barr virus oncoprotein latent membrane protein 1 (LMP1) mimics an activated CD40 receptor. In order to investigate the extent of similarity between LMP1 and CD40 functions in vivo, we analyzed the cytoplasmic signaling properties of LMP1 and CD40 in B cells in a directly comparable manner. For this purpose, we generated transgenic mice expressing either LMP1 or a chimeric LMP1CD40 molecule, which constitutively activates the CD40 pathway, under the control of the CD19 promoter. LMP1 and LMP1CD40 were expressed at similar levels in a B-lymphocyte specific manner. Similar to LMP1, LMP1CD40 suppressed germinal center (GC) formation and antibody production in response to thymus-dependent antigens, albeit to a greater extent than LMP1. Furthermore, the avidity of the antibodies produced against thymus-dependent antigens was lower for LMP1CD40 transgenic mice than for wild-type and LMP1 transgenic mice. GC suppression was linked to the ability of LMP1CD40 and LMP1 to downregulate mRNA and protein levels of BCL6 and to suppress the activity of the BCL6 promoter. In contrast to LMP1, LMP1CD40 caused an upregulation of CD69, CD80, and CD86 in B cells and a dramatic increase in serum immunoglobulin M. In addition, LMP1CD40 but not LMP1 transgenic mice had elevated numbers of marginal-zone B cells and increased populations of polymorphonuclear cells and/or neutrophils. Consistent with these findings, LMP1CD40 but not LMP1 transgenic mice showed signs of spontaneous inflammatory reactions and the potential for autoimmunity. PMID- 15542675 TI - Comprehensive cross-clade neutralization analysis of a panel of anti-human immunodeficiency virus type 1 monoclonal antibodies. AB - Broadly neutralizing monoclonal antibodies (MAbs) are potentially important tools in human immunodeficiency virus type 1 (HIV-1) vaccine design. A few rare MAbs have been intensively studied, but we still have a limited appreciation of their neutralization breadth. Using a pseudovirus assay, we evaluated MAbs from clade B infected donors and a clade B HIV(+) plasma against 93 viruses from diverse backgrounds. Anti-gp120 MAbs exhibited greater activity against clade B than non B viruses, whereas anti-gp41 MAbs exhibited broad interclade activity. Unexpectedly, MAb 4E10 (directed against the C terminus of the gp41 ectodomain) neutralized all 90 viruses with moderate potency. MAb 2F5 (directed against an epitope adjacent to that of 4E10) neutralized 67% of isolates, but none from clade C. Anti-gp120 MAb b12 (directed against an epitope overlapping the CD4 binding site) neutralized 50% of viruses, including some from almost every clade. 2G12 (directed against a high-mannose epitope on gp120) neutralized 41% of the viruses, but none from clades C or E. MAbs to the gp120 V3 loop, including 447 52D, neutralized a subset of clade B viruses (up to 45%) but infrequently neutralized other clades ( 80 g ethanol/day throughout adulthood) and four matched controls (<20 g/day) were generated. Two-dimensional electrophoresis was performed in triplicate on alcoholic and control samples and the resultant protein profiles analyzed for differential expression. Overall, 182 proteins differed by the criterion of twofold or more between case and control samples. Of these, 139 showed significantly lower expression in alcoholics, 35 showed significantly higher expression, and 8 were new or had disappeared. To date, 63 proteins have been identified using MALDI-MS and MS-MS. The finding that the expression level of differentially expressed proteins is preponderantly lower in the alcoholic brain is supported by recent results from parallel studies using microarray mRNA transcript. PMID- 15542696 TI - No association found between the type 1 sigma receptor gene polymorphisms and methamphetamine abuse in the Japanese population: a collaborative study by the Japanese Genetics Initiative for Drug Abuse. AB - It has been suggested that individual genetic factors are involved in susceptibility to drug dependence and the manifestation of drug-induced psychosis. The aim of this study was to examine the relation between methamphetamine abusers/psychosis and the type 1 sigma receptor gene polymorphisms. Subjects comprised 143 MAP abusers and 181 healthy controls. Two polymorphisms in the type 1 sigma receptor gene, GC-241-240TT and A61C (Gln2Pro), were examined in the present study. No significant differences were observed in either polymorphism between healthy controls and MAP abusers/psychosis. In the subgroup analyses, the rate of CC genotype of A61C tended to be higher in MAP patients who had experienced spontaneous relapse without MAP use than in those who had not (P = .06, OR = 3.02 95%CI = 0.92-9.92). However, the level of this significant trend did not remain after the Bonferroni's multiple correction. This study suggests that type 1 sigma receptor gene is unlikely to play a major role in substance abuse liability and/or the development of MAP psychosis. PMID- 15542697 TI - No association is found between the candidate genes of t-PA/plasminogen system and Japanese methamphetamine-related disorder: a collaborative study by the Japanese Genetics Initiative for Drug Abuse. AB - In the central nervous system, tissue-plasminogen activator (t-PA)/plasmin system is involved in long-term synaptic plasticity and remodeling, and participates in rewarding effects of methamphetamine (MAP), by acutely regulating MAP-induced dopamine release in the nucleus accumbens. The aim of this study was to examine the relationships between the patients with MAP abusers/psychosis and the t PA/plasminogen system genes. Subjects comprised 185 MAP abusers and 288 healthy controls. Four polymorphisms in the t-PA, plasminogen activator inhibitor, and plasminogen genes were examined in the present study. No significant differences were observed in each polymorphism between healthy controls and MAP abusers/psychosis. This study suggests that t-PA/plasminogen system is unlikely to be a major contributor to the substance abuse liability and/or the development of MAP psychosis. PMID- 15542698 TI - Association studies of neurotransmitter gene polymorphisms in alcoholic Caucasians. AB - Ethanol enhances mesolimbic/cortical dopamine activity in reward and reinforcement circuits. We investigated the hypothesis that risk for alcoholism may be mediated by genes for neurotransmitters associated with the dopamine reward system as well as genes for enzymes involved in ethanol metabolism. DNA was extracted from brain tissue collected at autopsy from pathologically characterized alcoholics and controls. PCR-based assays showed that alcoholism was associated with polymorphisms of the dopamine D2 receptor (DRD2) TaqI B (P = .029) and the GABAA-beta2 subunit C1412T (P = .012) genes, but not with the glutamate receptor subunit gene NMDAR2B (366C/G), the serotonin transporter gene (5HTTL-PR), the dopamine transporter gene DAT1(SLC6A3), the dopamine D2 receptor gene DRD2 TaqI A, or the GABAA alpha1(A15G), alpha6(T1519C), and gamma2(G3145A) subunit genes. The glial glutamate transporter gene EAAT2 polymorphism G603A was associated with alcoholic cirrhosis (P = .048). The genotype for the most active alcohol dehydrogenase enzyme ADH1C was associated with a lower risk of alcoholism (P = .026) and was less prevalent in alcoholics with DRD2TaqIA2/A2 (P = .047), GABAA-beta2 1412C/C (P = .01), or EAAT2 603G/A (P = .022) genotypes. Combined DRD2TaqI A or B with GABAA-beta2 or EAAT2 G603A genotypes may have a concerted influence in the predisposition to alcoholism. PMID- 15542700 TI - Serial analysis of gene expression in methamphetamine- and phencyclidine-treated rodent cerebral cortices: are there common mechanisms? AB - Pharmacological actions of methamphetamine (METH) and phencyclidine (PCP) are different, but both of them can induce similar psychiatric disorders including abuse, intoxication, withdrawal, and psychotic symptoms like those of schizophrenia. These mental disorders are caused not only by their direct pharmacological effects, but also by secondary brain damage containing gene expression changes. In order to broadly grasp these alterations, we used serial analysis of gene expression (SAGE), a transcriptome analysis. We analyzed three cDNA libraries from cerebral cortices of saline (1 mL/kg)-, METH (4 mg/kg)-, or PCP (10 mg/kg)-treated Wistar rats (one hour after i.p. administration). The numbers of total tags were about 50,000 in each library, and approximately 18,000 kinds of tags were identified respectively. From the comparisons of three groups, we found both METH- and PCP-reactive genes. Upregulated genes contained calmodulin 2, stromal cell-derived factor receptor 1, brain-specific angiogenesis inhibitor 1-associated protein 2, ras homologue enriched in brain, basigin and thyrotropin-releasing hormone receptor. Downregulated genes contained lipocalin 2, aldolase A, importin 13, fatty acid binding protein 3, and glycine receptor alpha2 subunit. These data suggest important clues of common molecular basis in METH- and PCP-related psychiatric disorders. PMID- 15542699 TI - Molecular mechanisms underlying the rewarding effects of cocaine. AB - The initially surprising observation that cocaine retains its rewarding effects in dopamine transporter (DAT) knockout (KO) mice led our laboratory to examine the effects of deletion of other monoaminergic genes on cocaine reward. Our initial approach to this problem was to combine DAT KO mice with serotonin transporter (SERT) KO mice to make combined DAT/SERT KO mice. The combination of these knockouts eliminates cocaine reward as assessed in the conditioned place preference (CPP) paradigm. We have also identified evidence that, in the absence of DAT, there is greater participation in cocaine reward by serotonin (SERT) and norepinephrine (NET) transporters. Both NET and SERT blockers (nisoxetine and fluoxetine) produced significant CPPs in DAT KO mice, but not in wild-type (WT) mice. The striking elimination of cocaine CPP in combined DAT/SERT KO mice contrasts with effects that we have identified in combined NET/SERT knockout mice, which display increases in cocaine reward, and with recent reports that suggest that DAT/NET combined KOs retain substantial cocaine CPP. Overall, these studies indicate important requirements for several monoaminergic system genes to fully explain cocaine reward, in particular those expressed by dopamine and serotonin systems. PMID- 15542701 TI - Involvement of signal transduction cascade via dopamine-D1 receptors in phencyclidine dependence. AB - We investigated the molecular mechanisms of development to phencyclidine (PCP) induced rewarding effect by using tyrosine hydroxylase (TH) heterozygous (TH(+/ )) mice. PCP (8 mg/kg) induced the place preference in wild-type mice pretreated with PCP (10 mg/kg/day for 28 days). The place preference induced by PCP is attenuated by 6-hydroxydopamine, a dopaminergic neurotoxin, and (+) SCH-23390, a dopamine-D1 receptor antagonist, but not by DSP-4, a noradrenergic neurotoxin, and (-) sulpiride, a dopamine-D2 receptor antagonist. In TH(+/-) mice pretreated with PCP (10 mg/kg/day for 28 days), no PCP (8 mg/kg)-induced place preference was observed. In wild-type mice pretreated with PCP, the levels of cAMP, cAMP response element binding protein (CREB), and c-fos mRNA in the nucleus accumbens were increased. The levels of cAMP, CREB, and c-fos mRNA in the nucleus accumbens were not increased by the same treatment schedule of PCP in TH(+/-) mice. These findings suggest that changes in dopaminergic and/or cAMP signal cascades induced by repeated PCP treatment play an important role in the development of PCP induced rewarding effect. PMID- 15542702 TI - Specific gene expression and possible involvement of inflammation in methamphetamine-induced neurotoxicity. AB - To reveal specific gene expression in methamphetamine (METH) -induced dopamine neurotoxicity, temporal characteristics of METH-induced changes in gene expression in dopaminergic neuronal cells were examined using the cDNA array and the differential display method. A number of genes in the class of "trafficking & protein turnover," "metabolic pathways," "transmitters & receptors," and "growth factors, cytokines" were upregulated after the METH treatment in the cDNA array assay. Whereas, some genes related to trafficking & protein turnover and "modulators, effectors & intracellular transducers" were decreased by METH. Some proteins associated with synaptic vesicle transportation indeed up- or downregulated after the METH treatment. These data suggest that the protein trafficking and degradation system is involved in the dopaminergic cell death induced by METH. Furthermore, focusing on inflammatory reactions after METH injection, possible neuroprotective property of nonsteroidal anti-inflammatory drugs were examined against METH-induced neurotoxicity. Coadministration of NSAID with METH significantly attenuated striatal dopamine terminal degeneration and microgliosis induced by METH, suggesting that the protective effects are based on their inhibitory activity on production of cytokines and nitric oxides or their suppressive action against microglia activation. PMID- 15542703 TI - Profiling of methamphetamine-induced modifications of gene expression patterns in the mouse brain. AB - Recently described DNA microarray technology allows parallel screening of expression patterns and regulation of hundreds of thousands of genes. In the present study, we used a microarray to examine the gene expressions in the midbrains of mice sacrificed 24 h after completion of a 7-day treatment period consisting of a once-daily treatment with saline (SS), saline followed by a single 2 mg/kg of body weight dose of methamphetamine (METH) (S-METH), or repeated 2 mg/kg METH doses (M-METH) that produced sensitization and place preference (rewarding effect). We used the commercially available cDNA microarray. Approximately 80% of the assessed transcripts in the total brain reached the Affymetrix criteria for "present" and "changed," as well as displaying > or =1.5-fold differences in hybridization intensity difference values in a comparison of SS data to S-METH or M-METH data. S-METH gene expression changes were observed in both up- and down-regulation, with 13 transcripts upregulated and 13 downregulated, whereas the majority of M-METH gene expression changes were observed in down-regulation, with 5 transcripts upregulated and 21 downregulated. We identified several genes that altered expression in both the S-METH and M-METH groups: a transcription factor gene, cellular stress/molecular chaperones, and a cellular regulatory gene. PMID- 15542704 TI - Prefrontal abnormality of schizophrenia revealed by DNA microarray: impact on glial and neurotrophic gene expression. AB - DNA microarrays with isotope labeling from gene-specific primers enable sensitive detection of rare mRNAs, including neurotrophin and cytokine mRNAs in the brain. Using high-quality RNA from postmortem brains, gene-expression profiles covering 1373 genes were assessed in the dorsoprefrontal cortex of schizophrenic patients and compared with those of nonpsychiatric subjects. Statistical analysis of the DNA microarray data confirmed the findings of a previous GeneChip study by Hakak et al. (Proc. Natl. Acad. Sci. USA Vol. 98, pp. 4746-4751, 2001). The highest frequency of mRNA expression alterations occurred in oligodendrocyte- and astrocyte-related genes in the prefrontal cortex of schizophrenic patients, followed by the category for the genes for growth factors/neurotrophic factors and their receptors. Whether each mRNA signal represents the expression of the individual genes or homologous genes in the category remains to be determined, however. To control for potential medication effects on patients, RNA from cynomolgus monkeys that were treated with haloperidol for 3 months was also subjected to DNA microarray analysis. A few genes overlapped between the gene expression profiles of the monkeys and patients. The present profiling study suggests a potential biological link between abnormal neurotrophic signals and impaired glial functions in schizophrenic pathology. PMID- 15542705 TI - Changes in expression of the mouse homologues of KIAA genes after subchronic methamphetamine treatment. AB - Amphetamine abuse may be associated with adaptive changes in gene expression in the brain. In the present study, a newly developed cDNA array system comprising mouse KIAA (mKIAA) cDNA clones was used to examine the gene expression affected by chronic methamphetamine treatment. Approximately 800 mKIAA clones were blotted onto a nylon membrane and hybridized with 33P-labeled cDNA derived from mRNAs isolated from the whole brains of mice that had been treated daily with saline or methamphetamine (2 mg/kg, i.p.) for 2 weeks. The arrays displayed robust hybridization for almost all transcripts. The results obtained from five experiments were averaged, each performed with triplicate samples. Several clones were chosen as positive candidates for methamphetamine-induced changes; however, only Per2 and mKIAA0099 genes showed a significantly increased expression (P < .05). Subsequently, with the focus on the period-related proteins, the expression of these proteins in various parts of the rat brain were assessed by immunoblot analysis. Chronic administration of methamphetamine (8 mg/kg, i.p., for 10 days) caused increased Per2 protein expression in the hippocampus. Interestingly, chronic methamphetamine treatment at a lower dose (4 mg/kg, i.p., for 10 days) induced an increase in SCN circadian oscillatory protein (SCOP) expression, also in the hippocampus. These data suggest that long-lasting alterations of the period-related gene expressions in the hippocampus might play an important role in methamphetamine addiction. PMID- 15542706 TI - Psychostimulant alters expression of DNA methyltransferase mRNA in the rat brain. AB - Methamphetamine (MAP), the most frequently abused substance in Japan, causes severe drug dependence and psychosis, similar to schizophrenia. It is suggested that long-term alterations in gene expression is related to MAP-induced brain dysfunction, including dependence and psychosis. DNA (cytosine-5) methyltransferase (Dnmt), a methylating enzyme of cytosine residues on CpG dinucleotides, plays an important role in X chromosome inactivation, genomic imprinting, and gene expression. Reelin is an extracellular matrix protein secreted by GABAergic interneurons. Heterozygous reeler mice that exhibit a 50% downregulation of reelin expression replicate the dendritic spine and GABAergic defects described in schizophrenia. DNA methylation plays an important role in the epigenetic modification of reelin expression. We previously found that MAP could alter expression of Dnmt1 mRNA in the rat brain. In this study, we examined the brain mRNA for Dnmt2 and reelin in MAP-treated Wistar rats. Acute MAP (4 mg/kg) treatment significantly decreased Dnmt2 mRNA by 27% to 39% in hippocampus dentate gyrus, CA1, and CA3 24 h after treatment, and significantly decreased reelin mRNA by 28% in frontal cortex 3 h after treatment. These results suggest that (1) MAP can alter DNA methylation as well as expression of genes in these brain regions, and (2) decrease in reelin mRNA in the frontal cortex is similar to heterozygous reeler mice, which might be related to schizophrenia-like psychotic symptoms of MAP psychosis. PMID- 15542707 TI - Effect of methamphetamine on cognition and repetitive motor behavior of mice deficient for dopamine D2 and D3 receptors. AB - Mice deficient for dopamine D2 and D3 receptors exhibit blunted D(1)-receptor responses to agonist stimulation. This blunted D1-receptor activity is prominent in the medial prefrontal cortex (mPFC) and results in a significantly impaired performance of the mutants in a test for spatial working memory. A single dose of methamphetamine (METH; 5 mg/kg i.p.), however, elicits a long-lasting increase in agonist-stimulated D1 receptor activity in the mPFC. In D2 mutants, this increase reaches wild-type levels, and the working memory of METH-treated mutants is completely rescued. In D3 mutants, however, the METH-induced increase in D1 receptor activity remains below wild-type levels and does not result in improved working memory performance. D2 and D3 mutants also differ in their locomotor responses to METH. Repeated administration of this drug (5 mg/kg administered three times at 2-h intervals) leads to a transition from horizontal hyperlocomotion to excessive orofacial stereotypy (taffy pulling) only in wild type and D3 mutants. In both genotypes, this transition is accompanied by a change in the relative ratios of striatal neuronal activation in two neurochemically distinct compartments, with striosomal neuronal activation exceeding that of the striatal matrix during stereotypy. Both the stereotypic response to METH and the associated predominant activation of neurons located in striosomes require D2-receptor expression. These studies indicate a differential requirement for D1- and D2-like receptor activation in mediating the effects of METH on cognitive and motor function. PMID- 15542708 TI - Differential response of nNOS knockout mice to MDMA ("ecstasy")- and methamphetamine-induced psychomotor sensitization and neurotoxicity. AB - It has been shown that mice deficient in neuronal nitric oxide synthase (nNOS) gene are resistant to cocaine-induced psychomotor sensitization and methamphetamine (METH)-induced dopaminergic neurotoxicity. The present study was undertaken to investigate the hypothesis that nNOS has a major role in dopamine (DA)- but not serotonin (5-hydroxytryptamine; 5-HT)-mediated effects of psychostimulants. The response of nNOS knockout (KO) and wild-type (WT) mice to the psychomotor-stimulating and neurotoxic effects of 3,4 methylenedioxymethamphetamine (MDMA; "Ecstasy") and METH were investigated. Repeated administration of MDMA for 5 days resulted in psychomotor sensitization in both WT and nNOS KO mice, while repeated administration of METH caused psychomotor sensitization in WT but not in KO mice. Sensitization to both MDMA and METH was persistent for 40 days in WT mice, but not in nNOS KO mice. These findings suggest that the induction of psychomotor sensitization to MDMA and METH is NO independent and NO dependent, respectively, while the persistence of sensitization to both drugs is NO dependent. For the neurochemical studies, a high dose of MDMA caused marked depletion of 5-HT in several brain regions of both WT and KO mice, suggesting that the absence of the nNOS gene did not afford protection against MDMA-induced depletion of 5-HT. Striatal dopaminergic neurotoxicity caused by high doses of MDMA and METH in WT mice was partially prevented in KO mice administered with MDMA, but it was fully precluded in KO mice administered with METH. The differential response of nNOS KO mice to the behavioral and neurotoxic effects of MDMA and METH suggests that the nNOS gene is required for the expression and persistence of DA-mediated effects of METH and MDMA, while 5-HT-mediated effects of MDMA (induction of sensitization and 5-HT depletion) are not dependent on nNOS. PMID- 15542709 TI - Methamphetamine and brain histamine: a study using histamine-related gene knockout mice. AB - The central histamine (HA) neurons that originate from the posterior hypothalamus modulate a variety of physiological functions. In order to investigate the roles of brain histaminergic neuron system in the behavioral effects of methamphetamine (METH), we administrated METH repeatedly to L-histidine decarboxylase (HDC)-, histamine H1 receptor-, H2 receptor-gene knockout (KO) mice, H1/H2 receptor-gene double KO mice, and wild type (WT) mice corresponding to each of them, and we measured locomotor activities. We also measured the contents of monoamines and amino acids in the brain of HDC-gene KO and WT mice after a single administration of METH. METH-induced locomotor hyperactivity and the development of behavioral sensitization were facilitated more in the HDC-gene KO mice and H1/H2 gene double KO mice than the WT mice, suggesting that brain histamine has an inhibitory effect on the METH action through both H1 and H2 receptors. In addition, neurochemical study suggested the involvement of the GABAergic neuron system in the inhibitory effect of brain histamine. PMID- 15542710 TI - Nootropic nefiracetam inhibits proconvulsant action of peripheral-type benzodiazepines in epileptic mutant EL mice. AB - Piracetam and structurally related nootropics are known to potentiate the anticonvulsant effects of antiepileptic drugs. It remains to be seen, however, whether these nootropics inhibit proconvulsant actions of many toxic agents including Ro 5-4864, a specific agonist for peripheral-type benzodiazepine receptors (PBR). The present study was designed to address this issue using EL mice, an animal model of epilepsy. In behavioral pharmacological experiments, EL mice were highly susceptible to convulsions induced by Ro 5-4864 (i.p.) in comparison with nonepileptic DDY mice. Nefiracetam administered orally to EL mice inhibited spontaneous seizures. In DDY mice, convulsions induced by Ro 5-4864 were prevented by nefiracetam when administered by i.v. injection. Aniracetam (i.v.) was partially effective, but piracetam and oxiracetam were ineffective as anticonvulsants. Binding assay for brain tissues revealed a higher density of mitochondrial PBR in EL mice compared with DDY mice. Binding of the PBR ligands Ro 5-4864 to either EL or DDY mouse brain was inhibited by micromolar concentrations of these nootropic agents in the sequence of nefiracetam > aniracetam >> oxiracetam, piracetam. This rank order is identical to potency as anticonvulsants. These data suggest that nefiracetam may prevent toxic effects of PBR agonists through interacting with PBR. PMID- 15542711 TI - Food-reinforced operant behavior in dopamine transporter knockout mice: enhanced resistance to extinction. AB - Dopamine (DA) plays roles in circuits that are important for brain reward and in striatal brain regions that are important for certain types of habit learning. These processes in wildtype, heterozygous, and homozygous dopamine transporter knockout (DAT-KO) mice, which were mildly food deprived and allowed to make nose poke responses for food reinforcement, were studied. The mice were given 20-min sessions of daily (a) baseline exposure to the operant chambers, (b) acquisition of nose-poke responses in which responses were reinforced under a fixed ratio (FR5) schedule, (c) a progressive ratio schedule in which the number of responses required to obtain food was gradually increased, and (d) extinction of responses in which nose pokes were not followed by food. Neither heterozygous nor homozygous DAT-KO mice differed from their wildtype litter mates in the number of nose pokes displayed during baseline exposures to the chambers, the number of sessions required for acquisition, the number of responses under the FR5 schedule, or the number of responses under the progressive ratio schedule. Interestingly, however, in the five extinction sessions in which food was no longer delivered by nose poking, homozygous DAT-KO mice exerted significantly more responses than mice of either of the other two genotypes. These lines of evidence suggest a greater resistance of DAT-KO mice to the elimination of the response and support roles of dopaminergic systems in habit memory. PMID- 15542712 TI - Psychostimulants and vesicle trafficking: a novel mechanism and therapeutic implications. AB - The monoamine vesicular transporter 2 (VMAT-2) has been associated with dopamine (DA) sequestration and protection against neurodegeneration caused by the intracellular oxidation of this monoamine. The data presented herein suggest that methylphenidate treatment enhances the amount of VMAT-2 protein and possibly its activity in the presynaptic cytosol, where it is able to increase the sequestration of DA and likely protect against its instability. In contrast, methamphetamine (METH) has an opposite effect on cytosolic VMAT-2 resulting in degradation of DA terminals. The fact that posttreatment of methylphenidate after a neurotoxic regimen of METH protects against resulting loss of DA parameters suggests that treatment with methylphenidate, or other DA transporter blockers, may be protective against degenerative disorders of DA pathways, such as Parkinson's disease. PMID- 15542713 TI - Substituted amphetamines that produce long-term serotonin depletion in rat brain ("neurotoxicity") do not decrease serotonin transporter protein expression. AB - Administration of high-dose D-fenfluramine (D-FEN) or parachloroamphetamine (PCA) produces long-lasting decreases in serotonin transporter (SERT) binding and tissue levels of serotonin (5-HT) in rat forebrain. These changes have been viewed as evidence for 5-HT neurotoxicity, but few studies have measured SERT protein levels. Thus, in the present study we determined the effect of high-dose D-FEN or PCA, administered according to a "neurotoxic" dosing regimen, on the density of SERT sites using ligand binding methods and on SERT protein levels using Western blots. Rats were sacrificed 2 days and 2 weeks after administration of drug or saline. The density of SERT was determined in homogenates of caudate and whole brain minus caudate. d-FEN and PCA decreased SERT binding by 30 to 60% in both tissues and at both time points. Similarly, D-FEN and PCA administration profoundly decreased tissue 5-HT and 5-HIAA in frontal cortex. Despite the large decreases in SERT binding and depletion of tissue 5-HT that occurred with d-FEN administration, SERT protein expression, as determined by Western blot analysis, did not change in either tissue or time point. PCA administration decreased SERT protein by about 20% only at the 2-day point in the caudate. Drug treatments did not change expression of glial fibrillary acidic protein (GFAP), a hallmark indicator of neuronal damage, in whole brain minus caudate in the 2-week group. These results support the hypothesis that D-FEN- and PCA-induced decreases in tissue 5-HT and SERT binding sites reflect neuroadaptive changes rather than neurotoxic effects. PMID- 15542714 TI - Similarities between methamphetamine toxicity and proteasome inhibition. AB - The monoamine neurotoxin methamphetamine (METH) is commonly used as an experimental model for Parkinson's disease (PD). In fact, METH-induced striatal dopamine (DA) loss is accompanied by damage to striatal nerve endings arising from the substantia nigra. On the other hand, PD is characterized by neuronal inclusions within nigral DA neurons. These inclusions contain alpha-synuclein, ubiquitin, and various components of a metabolic pathway named the ubiquitin proteasome (UP) system, while mutation of genes coding for various components of the UP system is responsible for inherited forms of PD. In this presentation we demonstrate for the first time the occurrence of neuronal inclusions in vivo in the nigrostriatal system of the mouse following administration of METH. We analyzed, in vivo and in vitro, the shape and the fine structure of these neuronal bodies by using transmission electron microscopy. Immunocytochemical investigation showed that these METH-induced cytosolic inclusions stain for ubiquitin, alpha-synuclein, and UP-related molecules, thus sharing similar components with Lewy bodies occurring in PD, with an emphasis on enzymes belonging to the UP system. In line with this, blockade of this multicatalytic pathway by the selective inhibitor epoxomycin produced cell inclusions with similar features. Moreover, using a multifaceted pharmacological approach, we could demonstrate the need for endogenous DA in order to form neuronal inclusions. PMID- 15542715 TI - Antagonists of the neurokinin-1 or dopamine D1 receptors confer protection from methamphetamine on dopamine terminals of the mouse striatum. AB - Methamphetamine (METH) is a highly addictive compound that induces toxicity of the dopamine (DA) terminals of the neostriatum. Exposure to METH induces long term deficits in dopamine transporter (DAT) and tyrosine hydroxylase (TH) levels as well as induction of glial fibrillary acidic protein (GFAP) in the caudate putamen (CPu) and the nucleus accumbens (NAc). The primary effect of exposure to METH is elevation of the level of extracellular DA; therefore, we assessed the role of the DA D1 receptor (D1R) and neurokinin-1 receptor (NK-1R) on the expression of toxicity. METH was injected intraperitoneally (10 mg/kg) four times at 2-h intervals (an acute toxic dose), and the mice were sacrificed three days after the treatment. Exposure to METH resulted in marked reduction of DAT sites (reduced to 30 and 21% relative to control in medial and lateral aspects of the CPu) assessed by binding of [125I]RTI-121 by autoradiography or Western blot analysis. Pretreatment with the nonpeptide NK-1R antagonist WIN-51,708 (10 mg/kg) 30 min prior to the first and fourth injections of METH prevented the loss of DAT sites of the CPu. Moreover, pretreatment with WIN-51,708 also prevented the reduction of TH levels induced by METH as well as the induction of GFAP in astrocytes. Pretreatment with the D1R antagonist SCH-23390 (0.25 mg/kg) 30 min before the first and fourth injections of METH conferred partial protection on DAT sites of the CPu. These results demonstrate that receptors postsynaptic to the DA terminals of the CPu are needed in order to express the neurotoxic effects of METH on integral components of the DA terminals of the nigrostriatal projection. PMID- 15542716 TI - Effects of repeated low doses of MDMA on EEG activity and fluoro-jade B histochemistry. AB - The psychostimulant 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") is an amphetamine derivative that is widely abused. In previous studies, depending on the animal species, neurotoxicity has been demonstrated for either serotonin (5 HT) or/and dopamine (DA) nerve endings. These studies focused on the basal ganglia circuitry; however, in humans chronic abuse of MDMA often results in neurological symptoms that last after MDMA withdrawal and are not related to the extrapyramidal system such as electroencephalographic (EEG) abnormalities and cognitive impairment. These alterations might be due to the concomitant intake of other illicit compounds, the consequence of MDMA-induced hyperthermia, or to a primary neurotoxicity directed to extrastriatal regions. These observations call for a more in-depth analysis on the potential involvement of brain areas outside the basal ganglia in the toxic effects induced primarily by MDMA. In the present study, we treated C57Black mice chronically (25 days) with daily injections of MDMA (2.5 mg/kg). During treatments, mice were monitored in order to detect behavioral modifications, and epidural electrodes were installed to perform EEG recording. Behavioral data showed a sensitization as measured by locomotor activity, which related to progressive and long-lasting EEG changes and neuronal degeneration within the hippocampus. PMID- 15542717 TI - Effects of "Legal X" piperazine analogs on dopamine and serotonin release in rat brain. AB - 3,4-Methylenedioxymethamphetamine (MDMA) is a popular illicit drug that evokes transporter-mediated release of serotonin (5-HT) and dopamine (DA) from nerve cells. Recently, drug users have ingested combinations of the piperazine analogs, 1-benzylpiperazine (BZP) and 1-(m-trifluoromethylphenyl)piperazine (TFMPP), in an attempt to mimic the subjective effects of MDMA. In the present study, we compared neurochemical effects of MDMA, BZP, and TFMPP in rat brain. The ability of MDMA, BZP, and TFMPP to stimulate efflux of [3H]5-HT and [3H]MPP+ (a DA transporter substrate) was determined in vitro using release assays in synaptosomes. The ability of these drugs to increase extracellular 5-HT and DA in vivo was assessed using intracranial microdialysis in nucleus accumbens. MDMA stimulated transporter-mediated release of 5-HT (EC50 = 58 nM) and MPP+ (EC50 = 119 nM). BZP was a selective releaser of MPP+ (EC50 = 175 nM), whereas TFMPP was a selective releaser of 5-HT (EC50 = 121 nM). MDMA injections (1 and 3 mg/kg, i.v.) increased dialysate 5-HT and DA in a dose-related manner, but actions on 5 HT were predominant. BZP (3 and 10 mg/kg, i.v.) elevated dialysate DA and 5-HT, while TFMPP (3 and 10 mg/kg, i.v.) elevated only 5-HT. The coadministration of BZP plus TFMPP (BZP/TFMPP) produced marked elevations in extracellular 5-HT and DA that mirrored the effects of MDMA. At the high dose of BZP/TFMPP (10 mg/kg, i.v.), the rise in dialysate DA exceeded the summed effects of the drugs alone. Our results support the hypothesis that the BZP/TFMPP combination mimics the neurochemical mechanism of MDMA, providing a basis for recreational use of these agents. Additionally, the findings suggest possible drug-drug synergism when piperazine drugs are coadministered at high doses. PMID- 15542718 TI - Pro- and anti-addictive neurotrophic factors and cytokines in psychostimulant addiction: mini review. AB - Drug addiction is defined as a chronically relapsing disorder that is characterized by compulsive drug taking, inability to limit the intake, and intense drug craving. While the positive reinforcing effects of psychostimulants such as cocaine and amphetamines depend on the mesocorticolimbic dopamine system innervating nucleus accumbens, chronic drug exposure causes stable changes in the structure and function of the brain that may underlie the long-lived behavioral abnormalities in drug addiction. Recent evidence has suggested that various neurotrophic factors and cytokines are involved in the effects of psychomotor stimulants, suggesting that these factors play a role in drug addiction. In this article, a role of neurotrophic factors and cytokines in psychostimulant addiction is discussed. PMID- 15542719 TI - Developmental and genetic influences upon gender differences in methamphetamine induced nigrostriatal dopaminergic neurotoxicity. AB - The gonadal steroid hormone estrogen (E) may play an important role in sex differences in methamphetamine (MA)-induced neurotoxicity of the nigrostriatal dopaminergic (NSDA) system because E can serve as a neuroprotectant in female, but not male, mice. Gonadal steroid hormones also exert important organizational/developmental effects upon the brain at critical developmental periods. In Part 1 we assessed whether organizational (neonatal) or developmental (prepubertal) effects of gonadal steroids would alter gender/E-dependent neuroprotection of MA-induced NSDA neurotoxicity. Attempts to feminize male mice by gonadectomy at either the neonatal or prepubertal period failed to enable E to function as a neuroprotectant within the adult male mouse. Attempts to masculinize the female by testosterone administration at the neonatal period did not abolish the capacity for E to function as a neuroprotectant. However, prepubertal gonadectomy of female mice did disrupt E's capacity to serve as a neuroprotectant. These results suggest that genetic sex may prove the primary determinant for the sex differences observed in response to MA-induced NSDA neurotoxicity. In Part 2 we examined whether gender differences in response to MA induced NSDA neurotoxicity would interact with a specific genetic alteration in a neurotrophic factor, brain-derived neurotrophic factor (BDNF). Female and male mice that were either deficient (+/- BDNF) or overexpressing (DBH:BDNF+) BDNF were treated with MA. Sex differences in MA-induced NSDA neurotoxicity remained present in +/- BDNF mice and were less severe as compared with their wild-type controls. A similar result was obtained in mice that overexpress BDNF, with female and mutant mice showing less NSDA neurotoxicity. In both BDNF-deficient mice and mice that overexpress BDNF, the relative degree of MA-induced NSDA neurotoxicity was lower in males. Taken together, these results suggest that a selective alteration in BDNF expression offers some neuroprotective potential against MA-induced NSDA neurotoxicity, and the relative degree of this neuroprotection may interact with the gender of the subject. PMID- 15542720 TI - Comparative effects of NO-synthase inhibitor and NMDA antagonist on generation of nitric oxide and release of amino acids and acetylcholine in the rat brain elicited by amphetamine neurotoxicity. AB - The aim of this study was to clarify the role of nitric oxide (NO) and lipid peroxidation (LPO) processes as well as the contribution of various neurotransmitters in pathophysiological mechanisms of neurotoxicity induced by amphetamine (AMPH). NO level was determined directly in brain tissues using electron paramagnetic resonance spectroscopy technique. The content of the products of lipid peroxidation (LPO) was measured spectrophotometrically as thiobarbituric acid reactive species (TBARS). The output of neurotransmitter amino acids (glutamate, aspartate, and GABA) and acetylcholine (ACH) was monitored in nucleus accumbens (NAc) by push-pull technique with HPLC detection. Repeated, systemic application of AMPH elevated striatal and cortical NO generation and LPO production. Moreover, administration of AMPH led to a marked and long-lasting increase of ACH release. Surprisingly, while glutamate output was not affected, aspartate release was enhanced 30 to 50 min after each AMPH injection. The release rate of GABA was also elevated. The selective NO-synthase inhibitor 7-nitroindazole (7-NI) was highly effective in abating the rise in the neurotransmitter release induced by the AMPH. The NOS inhibitor also abolished the increase of NO generation produced by AMPH, but did not influence the intensity of LPO elicited by the AMPH administration. Pretreatment with the noncompetitive NMDA receptor antagonist dizocilpine (MK-801) completely prevented increase of NO generation and TBARS formation induced by multiple doses of AMPH. Dizocilpine also abolished the effect of the psychostimulant drug on the release of neurotransmitters ACH, glutamate, aspartate, and GABA in the NAc. Our findings suggest a key role of NO in AMPH-induced transmitter release, but not in the formation of LPO products. It appears that AMPH enhances release of ACH and neurotransmitter amino acids through increased NO synthesis and induces neurotoxicity via NO and also by NO-independent LPO. PMID- 15542721 TI - Effects of N-acetyl-L-cysteine on the reduction of brain dopamine transporters in monkey treated with methamphetamine. AB - Several lines of evidence suggest that oxidative stress might contribute to neurotoxicity in the dopaminergic nerve terminals after administration of methamphetamine (MAP). The present study undertakes to determine whether intravenous administration of N-acetyl-L-cysteine (NAC), a potent antioxidant drug, could attenuate the reduction of dopamine transporter (DAT) in the striatum of monkey brain after administration of MAP. Positron emission tomography (PET) studies demonstrated that repeated administration of MAP (2 mg/kg as a salt, four times at 2-h intervals) significantly decreased the accumulation of radioactivity in the striatum after intravenous administration of [11C]b-CFT (for DAT). In contrast, the binding of [11C]DASB to 5-hydroxytryptamine transporter (5-HTT) in the monkey brain was slightly decreased after the administration of MAP, although the difference was not statistically significant. The binding of [11C]SCH 23390 to dopamine D1 receptors in the striatum was also not altered after the administration of MAP. A bolus injection of NAC (150 mg/kg, i.v.) 30 min before administration of MAP and a subsequent continuous infusion of NAC (12 mg/kg/h, i.v.) over 8.5 h significantly attenuated the reduction of DAT in the monkey striatum 3 weeks after the administration of MAP. These results suggest that NAC could attenuate the reduction of DAT in the monkey striatum after repeated administration of MAP. Therefore, it is likely that NAC would be a suitable drug for treatment of neurotoxicity on dopaminergic nerve terminals related to chronic use of MAP in humans. PMID- 15542722 TI - Fos expression associated with the discriminative stimulus effects of methamphetamine in rats. AB - Methamphetamine, a psychostimulant drug, produces both acute psychomotor stimulation and long-lasting behavioral effects including addiction and psychosis. To identify anatomical substrates for the discriminative stimulus effects of methamphetamine in rats, we examined the drug discrimination associated c-Fos expression in the brains of rats that were trained to discriminate methamphetamine from saline under a two-lever fixed ratio (FR-20) schedule of food reinforcement. c-Fos expression in the brains of rats trained to discriminate methamphetamine from saline was significantly increased in the nucleus accumbens (NAc) and the ventral tegmental area (VTA) as compared with the expression in the control rats that were maintained under the FR-20 schedule, but no alternation was observed in other areas including the cerebral cortex, caudate putamen, substantia nigra, hippocampus, amygdala, and habenulla. Methamphetamine treatment in the trained rats caused a significant increase in c-Fos expression in the VTA, and a decrease in the NAc core, as compared to saline treatment. However, c-Fos expression in the NAc and VTA of rats that received chronic intermittent methamphetamine administration without discrimination training, did not differ from the expression in saline-treatment animals. These results suggest that the VTA and the NAc play an important role in the discriminative stimulus effects of methamphetamine. PMID- 15542723 TI - Augmentation of serotonin-induced inhibition of neuronal activity in the hippocampus following repeated treatment with methamphetamine. AB - Electrophysiological studies were performed to determine whether or not hippocampal serotonergic modulation was affected following repeated administration of methamphetamine (MAP). Rats were i.p. administered with MAP (5 mg/kg) or saline once a day for 5 days. Hippocampal slices were prepared at 24 h, 5 and 10 days after the final MAP or saline injection. The population spikes (PS) induced by stimulation of Schaffer collateral/commissural fibers were recorded in the hippocampal CA1 region. Application of serotonin (5-HT) via a bath perfusion system inhibited the PS in a concentration-dependent manner. At 24 h after the final injection, 10-microM 5-HT-induced inhibition of PS was not affected by MAP treatment. However, 5 days after the final injection, the inhibition by 5-HT (10 microM) of PS was significantly augmented in the MAP-treated group. Ten days after the final injection, this augmentation was not statistically significant compared with that of control group. 8-OH-DPAT, a 5-HT1A receptor agonist, inhibited PS, but the inhibition was not enhanced or reduced 5 days after the final MAP treatment. However, the enhancement of PS by RS 67333, a 5-HT4 receptor agonist, was attenuated 5 days after the MAP treatment. It was found that 5-HT induced inhibition of PS in the hippocampal CA1 region was potentiated 5 days after cessation of MAP, and this effect was suggested to be due to reduction of excitatory 5-HT4 receptor functions. PMID- 15542724 TI - The altered disposition of methamphetamine in the model of methamphetamine induced neurotoxicity. AB - Methamphetamine (METH) is a drug of abuse, causing neurotoxic effects in mammals. Many hypotheses have been proposed to explain the underlying mechanisms of METH induced toxicity, based on neurochemical/neuroanatomical changes. However, the pharmacokinetic properties of METH in the METH-induced neurotoxic model have not yet been evaluated. Thus, we investigated plasma and tissue levels of METH in the METH-induced neurotoxic model. As a result, when METH is administered multiply (5 mg/kg 4 times at 2-h intervals) in male Wistar rats, plasma METH levels at the third and forth injections were significantly higher than those at the first. The tissue distributions of METH in the brain as well as in the kidney were significantly decreased in the third injections, suggesting the importance of decreased transport of METH into tissues. Alternatively, one week after the establishment of METH-induced neurotoxicity, plasma levels of METH were back to normal, although METH levels in brain microdialysates were significantly higher than those in normal animals. These results suggest that the altered pharmacokinetic properties of METH, due to the abnormal membrane transport/disposition of METH into both central and peripheral tissues, might partially affect the emergence of METH-induced neurotoxicity. PMID- 15542725 TI - Social isolation stress significantly enhanced the disruption of prepulse inhibition in mice repeatedly treated with methamphetamine. AB - Repeated administration of methamphetamine (METH) causes reverse tolerance or behavioral sensitization in mice. However, the effects of social isolation stress on the METH-caused reverse tolerance have not been studied until now. The aim of this study was to investigate the effects of social isolation stress on METH caused reverse tolerance by examining the prepulse inhibition of startle response (PPI). PPI was tested in socially isolated and grouped mice after repeated METH injections. Locomotor activity and PPI were also examined just after a four-week isolation rearing period as a control experiment. After completing behavioral experiments, the mice were sacrificed, and the contents of monoamines, including histamine in the brain, were measured. Social isolation stress significantly lowered the locomotion and disrupted PPI. Repeated injections of METH enhanced the effects of social isolation on PPI. The content of dopamine and histamine significantly increased in the cortex, and the turnover rate of dopamine decreased significantly. These findings demonstrate that social isolation stress significantly enhances METH-induced behavioral sensitization and that the altered histaminergic neuron system might play an important role in METH-induced behavioral sensitization in addition to dopaminergic and serotoninergic neurotransmission. Our data suggest that social isolation is involved in the development of METH-induced psychosis, schizophrenia, and other related psychiatric disorders. PMID- 15542726 TI - Role of mitochondrial dysfunction in neurotoxicity of MPP+: partial protection of PC12 cells by acetyl-L-carnitine. AB - The damage to the central nervous system that is observed after administration of either methamphetamine (METH) or 1-methyl-4-phenylpyridinium (MPP+), the neurotoxic metabolite of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), is known to be linked to dopamine (DA). The underlying neurotoxicity mechanism for both METH and MPP+ seem to involve free radical formation and impaired mitochondrial function. The MPP+ is thought to selectively kill nigrostriatal dopaminergic neurons by inhibiting mitochondrial complex I, with cell death being attributed to oxidative stress damage to these vulnerable DA neurons. In the present study, MPP+ was shown to significantly inhibit the response to MTT by cultured PC12 cells. This inhibitory action of MPP+ could be partially reversed by the co-incubation of the cells with the acetylated form of carnitine, acetyl-L carnitine (ALC). Since at least part of the toxic action of MPP+ is related to mitochondrial inhibition, the partial reversal of the inhibition of MTT response by ALC could involve a partial restoration of mitochondrial function. The role carnitine derivatives, such as ALC, play in attenuating MPP+ and METH-evoked toxicity is still under investigation to elucidate the contribution of mitochondrial dysfunction in mechanisms of neurotoxicity. PMID- 15542727 TI - Nefiracetam attenuates methamphetamine-induced discriminative stimulus effects in rats. AB - Nefiracetam has been reported to increase intracellular cyclic AMP levels and enhance calcium channel current. Since the cAMP cascade is involved in the development of drug dependence, we investigated whether nefiracetam attenuates the methamphetamine (MAP)-induced discriminative responses in rats. Nefiracetam (50 mg/kg) inhibited MAP-induced discriminative responses. Furthermore, rolipram, raclopride, and L-745870, all of which can enhance cAMP activity, disrupted MAP paired lever press of rats. Nifedipine and neomycin, which are blockers of voltage-sensitive calcium channels (VSCCs), decreased MAP-induced discriminative responses. However, pretreatment of these VSCC blockers failed to affect the inhibitory effects of nefiracetam on MAP-induced discriminative responses. Our findings suggested that nefiracetam inhibits MAP-induced discriminative responses, which may be at least partly associated with the changes in intracellular cAMP levels. PMID- 15542728 TI - Clinical features of sensitization to methamphetamine observed in patients with methamphetamine dependence and psychosis. AB - Methamphetamine (METH) has been the most popular drug of abuse in Japan for more than 50 years, resulting in serious health and social issues. Most adult abusers in Japan consume only METH; multiple-substance abusers are rare. This unusual aspect of drug abuse makes it possible to observe clearly the sequential alteration of psychiatric symptoms induced by METH without modification by other illegal drugs. Clinical investigation reveals three core characteristics of METH abuse: (1) progressive qualitative alteration in mental symptoms from a nonpsychotic to a prepsychotic to a severely psychotic state; (2) enhanced vulnerability to relapse of psychosis; and (3) very long duration of the vulnerability to relapse. These findings indicate that the phenomenon of sensitization to METH develops during abuse and plays a key role in the susceptibility to and onset of psychosis and in the refractory process. Molecular findings using animal sensitization models may facilitate a better understanding of, and open the way for innovative therapies for, METH psychosis and also chronic schizophrenia. PMID- 15542729 TI - Neuromechanism of developing methamphetamine psychosis: a neuroimaging study. AB - The long-term use of methamphetamine (MAP) induces a psychotic state, called MAP psychosis. To understand the neuromechanisms of the persistent psychosis, we used SPECT, MR spectroscopy (MRS), and PET on the MAP users. The SPECT study showed a high incidence of multiple patchy deficits in cerebral blood flow among the users. The MRS study MAP users showed a significantly reduced ratio of creatine plus phosphocreatine (Cr + PCr)/choline-containing compounds (Cho) in the brain compared with the healthy control subjects. In addition, the reduction in the ratio of Cr + PCr/Cho was significantly correlated with the duration of MAP use and with the severity of residual psychiatric symptoms. PET revealed no significant differences between the ex-users of MAP and the healthy controls in the density of striatal dopamine D2 receptors. On the other hand, the density of dopamine transporter in the nucleus accumbens and caudate/putamen in the MAP users was significantly less compared with the controls. This reduction was significantly correlated with the length of use and severity of psychotic symptoms. These findings suggest that long-term use of MAP causes abnormal cerebral blood flow patterns, reduction of brain dopamine transporter density, and metabolite alteration, which may be closely related to a susceptibility to MAP psychosis. PMID- 15542730 TI - The role of noradrenergic and dopaminergic hyperactivity in the development of spontaneous recurrence of methamphetamine psychosis and susceptibility to episode recurrence. AB - The role of dopaminergic activity in susceptibility of methamphetamine (MAP) psychosis (flashbacks) to subsequent spontaneous recurrences was studied. Plasma monoamine metabolite levels were assayed in 23 flashbackers, of whom 10 experienced a single flashback, 8 exhibited subsequent flashbacks and 5 with the last episode; 18 nonflashbackers with a history of MAP psychosis; 9 subjects with persistent MAP psychosis; and 19 MAP user and 10 nonuser controls. All flashbackers had undergone frightening stressful experiences during previous MAP use. Their flashbacks were triggered by mild psychosocial stressors. Plasma norepinephrine (NE) levels increased with the increase in plasma levels of 3 methoxytyramine (3-MT), an index of dopamine release, during flashbacks in the 23 flashbackers. Of these, the 8 with subsequent episodes had markedly increased NE levels and increased 3-methoxytyramine levels during flashbacks. However, the 5 flashbackers with the last episode had moderately increased NE levels, and the 10 with a single episode displayed small increases in NE levels during flashbacks. Their 3-MT levels did not significantly differ from the levels in the control groups. Thus, increased DA release in addition to robust noradrenergic hyperactivity in response to mild psychosocial stressors may be important in susceptibility to subsequent flashbacks. PMID- 15542731 TI - A polymorphism of DRD2 gene and brain atrophy in methamphetamine psychosis. AB - Our group, Ujike et al., recently reported that the A1 allele of TaqI A polymorphism of the dopamine receptor D2 (DRD2) gene, associated with transient psychosis, significantly differs from that of patients with prolonged psychosis in methamphetamine psychosis. Therefore, we examined the association between the TaqI A polymorphism of the DRD2 gene and the brain MRI view for patients with methamphetamine psychosis. The subjects underwent brain MRI scans using the FLAIR method. Genotyping was performed by PCR-RFLP methods using genomic DNA extracted from peripheral blood by the phenol method. Ten subjects had the A1/A2 genotype, eleven subjects had the A2/A2 genotype, and no subject had the A1/A1 genotype. The domain size, including the thalamus and basal ganglia that were inside each side of the putamens, did not differ between the three groups (the A1/A2-group, the A2/A2-group, and the young healthy person group). In the comparison based on this domain, the temporal lobe tended to narrow in the A2/A2-group compared to the A1/A2-group (P = .06). The other domain (cerebrum, corpus callosum, etc.) showed no difference between the A1/A2-group and the A2/A2-group. It is suggested that in methamphetamine psychosis the TaqI A polymorphism not only regulates prolongation of psychosis symptoms but also influences the form of the temporal lobe. PMID- 15542732 TI - Gene polymorphisms of the mu opioid receptor in methamphetamine abusers. AB - In drug addiction, the opioid system is thought to mediate motivational effects through dopamine-independent mechanisms. We have investigated associations of the mu-opioid receptor gene (OPRM) variations with methamphetamine (MAP) dependence/psychosis. The allelic frequency of A118G (Asn40Asp) in exon 1 of ORPM was 45.3% in our control subjects, but only 7.5-25.8% in the Caucasian or African American population of previous studies. We have identified several novel polymorphisms in intron 1 and the 5' untranslated region (5'UTR) of OPRM. Polymorphisms in the functionally relevant 5' regulatory region of OPRM were different in our Japanese population from Caucasian or African-American populations. No significant differences between controls and MAP abusers were found in either genotype or allele frequency at any single nucleotide polymorphism (SNP) or (AC)n dinucleotide repeat in intron 1. A subdivision of our MAP group revealed that A118G of OPRM shows a significant association with MAP psychosis having latency less than three years. Further analysis should be capable of identifying associations between the OPRM variations and MAP dependence/psychosis. PMID- 15542734 TI - Role of Cdk5 in drug abuse and plasticity. AB - Neuronal plasticity serves as the basis for learning and memory in the adult brain. Contextual, motor, and reward-based learning are important in reinforcing survival behavior in animals. Most psychostimulant drugs of abuse target the dopaminergic reward system of the brain. Drugs of abuse cause long-standing cellular and molecular neuroadaptations in the brain. The neuronal protein kinase Cdk5 is emerging as an important player in the cellular and physiological responses to drugs of abuse. Substantial evidence indicates that Cdk5 controls dopamine neurotransmission through regulation of the protein phosphatase-1 inhibitor, DARPP-32. Furthermore, the morphological changes associated with chronic cocaine exposure are dependent on Cdk5. Thus, Cdk5 mediates cellular responses to psychostimulant drug-induced changes in dopamine signal transduction and cytoskeletal reorganization. In this regard, Cdk5, through its targeting of various substrates, integrates a number of intracellular pathways that are targeted by psychostimulant drugs. These studies and the emerging role of Cdk5 in various forms of neuronal plasticity are reviewed. PMID- 15542733 TI - Study of association between alpha-synuclein gene polymorphism and methamphetamine psychosis/dependence. AB - Methamphetamine (MAP) dissipates proton gradients across the membranes of synaptic vesicles, enhances cytoplasmic dopamine (DA) concentrations, and causes calcium-independent, nonvesicular DA release into synapses. MAP is taken into the cytosol by the dopamine transporter (DAT) on the synaptic terminals of DA neurons, and endogenous DA is concurrently released through the transporter by carrier exchange mechanisms, resulting in a robust increase in DA concentration in the synaptic clefts. The enhanced DA release through DAT by MAP is the main mechanism for the reinforcing effects of MAP. The complexes of alpha-synuclein and DAT facilitate membrane clustering of the DAT, thereby accelerating DA uptake in vitro. alpha-Synuclein has been shown to be overexpressed in the midbrain DA neurons of chronic cocaine abusers. The present study was performed to study the association between the alpha-synuclein gene polymorphisms and MAP psychosis/dependence in Japanese population. Since the T10A7 polymorphic site at the 5' end of the noncoding exon 1' in the alpha-synuclein gene is highly polymorphic, we analyzed the noncoding exon 1' and intron 1, including this polymorphic site by sequencing. We confirmed four single nucleotide polymorphisms (SNPs) within 1.38 kbp of the T10A7 polymorphic site. No significant difference was found in genotype or allele frequencies in the T10A7 polymorphic site between MAP psychotic/dependent and control subjects. We found significant association between three SNPs in the vicinity of this polymorphic site in intron 1 and MAP psychosis/dependence in female subjects, but not in males. These results suggest an association of the alpha-synuclein gene polymorphisms with MAP psychosis/dependence in our female subjects. Further analyses are necessary to clarify the gender difference, by using a larger sample size and/or different ethnic groups, as well as functional variations in the alpha-synuclein gene. PMID- 15542735 TI - Cocaine downregulates the expression of the mitochondrial genome in rat brain. AB - Analysis by differential display of genes induced in response to acute cocaine administration to rats revealed the significant downregulation of several mitochondrial genes in the cingulate cortex, including the subunits 1, 2, 4, 5, and 6 of NADH dehydrogenase and the subunit 2 of cytochrome c oxidase. Although the mechanism of the downregulation of expression of these mitochondrial genes by cocaine is presently not well understood, one can envisage that it involves an increased production of reactive oxygen species in cells of the cerebral cortex. PMID- 15542736 TI - Repetitive cocaine administration decreases neurogenesis in adult rat hippocampus. AB - Cocaine HCl (20 mg/kg) was administered to adult male rats to investigate the effects of cocaine on neurogenesis in the hippocampus. Proliferation of granule cells in the dentate gyrus was measured by in vivo labeling with 5-bromo-2' deoxyuridine (BrdU). Rats that received repetitive cocaine treatment for 14 days showed 26% fewer BrdU-positive cells relative to control rats, while no difference was observed in the rats that received a single injection of cocaine. Differentiation of newly born cells was not influenced. The present experiment is the first to demonstrate the influence of cocaine on hippocampal neurogenesis. These data suggest that the regulation of hippocampal neurogenesis may be involved in the emergence of certain symptoms of cocaine addiction, such as cognitive impairment and behavioral sensitization. PMID- 15542737 TI - CART peptides: modulators of mesolimbic dopamine, feeding, and stress. AB - CART peptides have been shown to be peptide neurotransmitters and endocrine factors in a series of cumulative studies over the past eight years or so. This brief review touches on three aspects of CART: CART as a mediator or modulator of mesolimbic dopamine, CART's regulation by glucocorticoids, and CART as a regulator of feeding, satiety, and body weight. There have been several recent reviews and publications on various aspects of CART peptides. These aspects include the sequence and numbering of the peptides, and their structure, processing, and roles in various physiologic processes. PMID- 15542738 TI - A possible genetic mechanism underlying individual and interstrain differences in opioid actions: focus on the mu opioid receptor gene. AB - Individual differences in responses to opioids limit effective pain treatment with these drugs. Identifying the mechanism could help to improve the analgesic effects of them. Since the molecular cloning of the mu opioid receptor (muOR) gene, substantial advances in opioid research have been made, including the discoveries that muOR plays a mandatory role in the analgesic effects of opioids and that the sequence of the muOR gene varies from one individual to another. It is conceivable that the differences in the muOR gene cause individual differences in opioid actions. The present review summarizes the recent advances made in research on human and mouse muOR genes and proposes that the variances in the 3' untranslated region (39-UTR) of the muOR gene might participate in the variability of the opioid response. PMID- 15542739 TI - Locus-specific involvement of anti-opioid systems in morphine tolerance and dependence. AB - Opioid tolerance and addiction could be discussed as two types of plasticity or counteradaptation, at the cellular level and through neuronal circuits. Cellular counteradaptation mechanisms include receptor desensitization through phosphorylation and endocytosis and through altered gene expression. The former mechanisms are related to the acute tolerance mechanisms, while the latter to chronic one. From current studies, it is known that various phosphorylation steps, such as protein kinase C (PKC) and G protein-coupled receptor (GPCR) kinase (GRK) regulate endocytosis. Of interest is that there are some differences in the physiological roles between opioid receptor endocytosis and other GPCR ones. Endocytosis of the opioid receptor is conceived as a recycling and resensitization step rather than the desensitization step. PKC phosphorylation inhibits endocytosis (PKC hypothesis). Therefore the PKC inhibitor attenuates acute analgesic tolerance. The agonist, which shows high-endocytosis stimulation, therefore makes less significant tolerance liability (RAVE hypothesis). Chronic tolerance is more likely related to the mechanisms through plastic modulation of neuronal circuits, where anti-opioidergic neurons are involved. The knockout mice lacking the receptors for anti-opioidergic nociceptin/orphanin FQ (N/OFQ) or glutamatergic neurons show weak or no morphine tolerance and dependence. As their gene expression or protein expression increases during chronic morphine treatments, we propose the hypothesis that the enhanced anti-opioid system may cause a counteradaptation to show tolerance and dependence. By a novel electroporation technique to deliver the receptor into the brain of knockout mice, we succeeded in determining the specific locus for the site of anti-opioid (through GluRepsilon1 or NR2A) action. All these results suggest that enhanced anti-opioid systems may contribute to the development of morphine tolerance and dependence, and their contributions could be brain locus specific. PMID- 15542740 TI - Involvement of glial glutamate transporters in morphine dependence. AB - There are several lines of evidence implying the involvement of the central glutamatergic system in morphine dependence. Extracellular glutamate released from nerve terminals is counterbalanced by glutamate transporters in neurons (EAAC1 and EAAT4) and glial cells (GLT-1 and GLAST), thereby modulating the glutamatergic system and protecting neurons from an excitotoxic action of glutamate. Here we show that a glial glutamate transporter GLT-1 could be involved in physical and psychological morphine dependence. By Northern blot analysis, the expression of glial glutamate transporter GLT-1, but not GLAST, mRNA was decreased in the striatum/nucleus accumbens (NAc) and thalamus of morphine-dependent rats. Subcutaneous administration of a glutamate transporter activator suppressed the development of physical morphine dependence and morphine induced conditioned place preference. Intracerebroventricular administration of a glutamate transporter inhibitor to morphine-dependent rats facilitated the expression of naloxone-precipitated morphine withdrawal-induced somatic signs and conditioned place aversion. Furthermore, gene transfer techniques using recombinant adenoviruses revealed that GLT-1 in the locus coeruleus and NAc shell plays inhibitory roles in physical and psychological morphine dependence, respectively. These findings may provide evidence that a glial glutamate transporter GLT-1 could be a new target for preventing physical and psychological morphine dependence. PMID- 15542742 TI - Pharmacological characterization of nociceptin/orphanin FQ receptors, a novel opioid receptor family, in the midbrain periaqueductal gray. AB - A fourth opioid receptor family was cloned and named after its endogenous ligand as nociceptin/orphanin FQ (N/OFQ) peptide (NOP) receptor. We have characterized several NOP receptor ligands pharmacologically at native NOP receptors of ventrolateral periaqueductal gray (vlPAG) neurons by investigating their interactions with N/OFQ in activating G protein-coupled inwardly rectifying K+ (GIRK) channels. They are listed here: (1) [Phe1Psi(CH2-NH)Gly2]N/OFQ(1-13)NH2, which was claimed to be the first selective antagonist of NOP receptors, is a partial agonist of NOP receptors in vlPAG neurons. (2) [Nphe1]N/OFQ(1-13)NH2 is a pure, selective, and competitive peptide antagonist of NOP receptors (pA2 value = 6.6). (3) CompB (J-113397) is a potent and selective nonpeptide antagonist of NOP receptors (pA2 = 8.4). (4) Naloxone benzoylhydrazone is a competitive NOP receptor antagonist but also a noncompetitive mu-opioid receptor antagonist. (5) Ro 64-6198, though being developed as a potent nonpeptide NOP receptor agonist, affected only part of vlPAG neurons and acted as a weak NOP receptor agonist. In Ro 64-6198-unresponsive neurons, N/OFQ activated GIRK channels through NOP receptors. (6) Nocistatin, a functional antagonist of N/OFQ in the spinal cord, did not affect the effect of N/OFQ in most of the recorded neurons. Our functional studies of NOP receptor ligands at native brain NOP receptors reveal that some of them act differently from those at expressed receptors of cell cultures. PMID- 15542741 TI - Negative regulation of opioid receptor-G protein-Ca2+ channel pathway by the nootropic nefiracetam. AB - It has recently been reported that nefiracetam, a nootropic agent, is capable of attenuating the development of morphine dependence and tolerance in mice. The mechanism of this antimorphine action is not clear. The present study was designed to address this issue using Xenopus oocytes expressing delta-opioid receptors, G proteins (G(i3alpha) or G(o1alpha)), and N-type (alpha1B) Ca2+ channels. Membrane currents through Ca2+ channels were recorded from the oocytes under voltage-clamp conditions. The Ca2+ channel currents were reduced reversibly by 40-60% in the presence of 1 microM leucine-enkephalin (Leu-Enk). The Leu-Enk induced current inhibition was recovered promptly by nefiracetam (1 microM), while control currents in the absence of Leu-Enk were not influenced by nefiracetam. A binding assay revealed that 3H-nefiracetam preferentially bound to the membrane fraction of oocytes expressing G(i3alpha). When delta-opioid receptors were coexpressed, the binding was significantly increased. However, an additional expression of alpha1B Ca2+ channels decreased the binding. The results suggest that nefiracetam preferentially binds to G(i3alpha) associated with delta opioid receptors, thereby inhibiting the association of G proteins with Ca2+ channels. In conclusion, nefiracetam negatively regulates the inhibitory pathway of opioid receptor-G protein-Ca2+ channel. PMID- 15542743 TI - Possible pharmacotherapy of the opioid kappa receptor agonist for drug dependence. AB - Because there are few efficacious medications for drug dependence, many clinical trials are being conducted in earnest to find such medications. Considerable evidence has shown that opioid kappa receptor agonists attenuate several behavioral responses induced by drugs of abuse. Although this raises the possibility that opioid kappa receptor agonists may be useful for the treatment of drug dependence on drugs of abuse, it has been previously reported that treatment with selective opioid kappa receptor agonists causes a psychotomimetic effect and dysphoria both in clinical studies and experimental animal models. As a result, we found the novel opioid kappa receptor agonist TRK-820, another chemical class of opioid kappa receptor agonist that has a morphinan scaffold unlike prototypical opioid kappa receptor agonists, by application of a modified message-address concept. TRK-820 showed high selectivity for an opioid kappa receptor, and strong agonistic activity in both in vitro and in vivo experiments. Like other opioid kappa receptor agonists, TRK-820 could markedly suppress the rewarding effects induced by morphine and cocaine and the discriminative stimulus effect of cocaine. Furthermore, TRK-820 attenuated the mecamylamine-precipitated nicotine-withdrawal aversion in a conditioned place preference paradigm. It is worthwhile to note that unlike prototypical opioid kappa receptor agonists, TRK 820 failed to produce a significant place aversion in rodents at doses that were sufficient to produce significant antinociception. Taken together, these findings indicate that TRK-820 may be useful for the treatment of drug dependence without any aversive effects. PMID- 15542744 TI - A single exposure to morphine induces long-lasting hyporeactivity of rat caudate putamen dopaminergic nerve terminals. AB - The long-lasting effects of exposure to drugs of abuse on the brain is a central theme in drug addiction research. This study was designed to evaluate whether enduring neurochemical adaptations within caudate putamen can be evoked by a single injection of a high dose of morphine. Rats were pretreated once with 10 mg/kg morphine. Seven days later the effect of another injection of 10 mg/kg morphine on total levels of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanilic acid (HVA) in caudate putamen was assessed in half the pretreated animals. An irreversible mu-opioid receptor antagonist, cloccinamox (C CAM; 0.1 mg/kg), significantly antagonized the elevation of the HVA/DA ratio, but not the elevation of the DOPAC/DA ratio induced by morphine in the caudate putamen from drug-naive animals. Pretreatment with morphine blunted changes in the HVA/DA ratio induced by another morphine challenge, but it had no effect on the DOPAC/DA ratio within the caudate putamen. Therefore, a single dose of 10 mg/kg morphine hampered nigrostriatal DA release and extraneuronal metabolism, mu opioid receptor mediated, on another 10 mg/kg morphine challenge. This confirms that the first exposure to morphine does not go without long-lasting neurochemical adaptations. PMID- 15542745 TI - Effects of L-745,870, a dopamine D4 receptor antagonist, on naloxone-induced morphine dependence in mice. AB - We examined whether dopamine D4 receptor is involved in morphine dependence in mice. Mice pretreated with morphine (10 mg/kg, s.c.) twice a day for 5 days showed withdrawal syndromes such as jumping, rearing, and forepaw tremors after the administration of naloxone (2 mg/kg, i.p.) on the sixth day. Such mice exhibited significant elevation of cAMP levels in the thalamus compared with the control mice. L-745,870 (1 mg/kg, i.p.), a selective dopamine D4 receptor antagonist, pretreated with morphine on the sixth day, significantly attenuated the severity of withdrawal syndromes and the increase in cAMP levels after the administration of naloxone. These results suggest that (1) the elevation of cAMP levels is involved in the expression of morphine-induced withdrawal syndromes, and (2) dopamine D4 receptor antagonists inhibit the expression of morphine induced withdrawal syndromes accompanied with biochemical changes in mice. Furthermore, dopamine D4 receptor antagonists may be useful drugs for attenuating the expression of morphine dependence. PMID- 15542746 TI - Recent trends of drug abuse in Japan. AB - Methamphetamine has been a continuous cause for concern for many years in Japan. Japan experienced the pandemics of methamphetamine abuse twice after World War II and is now facing the third pandemic. New data indicate that the abuse of drugs such as MDMA (ecstasy) and cannabis is rising at an alarming pace. An epidemiologic survey conducted by the National Research Institute reveals that drug abuse in Japan is still very low compared to that in other countries and regions; however, the current situation seems to be changing for the worse. Two major factors accelerate access to illicit drugs among young people. One is the presence of illicit drug-dealing groups, which indiscriminately offer drugs. The other is new technology such as the Internet and mobile phones. Strict law enforcement and intensive public awareness and prevention activities are indispensable in protecting young people from drug abuse. PMID- 15542747 TI - Drug abuse in China. AB - The drug problem in China reappeared in the late 1980s. At that time, most drug abusers used opium only, with heroin accounting for a very small proportion and its use being limited to border areas in the southwest and rural areas in the northwest. Beginning in the early 1990s, drug abuse spread quickly. The number of registered drug addicts increased from 70,000 in 1990 to one million by the end of 2002. Besides opioids, some "new" kinds of drugs including amphetamines and ketamine have penetrated the country through various channels since 1997. Drug abuse has caused severe problems for both the abusers and society. Three types of treatment settings for detoxification and rehabilitation are available: compulsory detoxification institutions, rehabilitation units through labor, and voluntary detoxification institutions run by sections of public security, justice, and health, respectively. Medical treatment is often combined with psychological counseling and physical training. Pharmaceutical therapy, traditional Chinese medicine, and nonpharmaceutical therapy are utilized for opioid addiction. Drug prevention, especially for young people, is emphasized. NGOs at different levels have all joined in this work. Intervention work was also started in regions where drug abuse and HIV/AIDS problems are serious. Drug abusers are helped to keep drug free after being released from detoxification settings by their communities. Different levels of institutions engaged in research on drug abuse are found in all parts of China. PMID- 15542748 TI - Drug problem in southeast and southwest Asia. AB - In 2002, the drug problem in Southeast and Southwest Asia was serious, particularly in the production of opium and heroin in Afghanistan, Myanmar, and Laos, the three largest producers of illicit opium in the world. The increasing illicit manufacture of ATS, particularly methamphetamine, in Southeast Asia, mainly in China and Myanmar, was also a major concern. Some reports indicated that ephedrine, used for illicitly producing methamphetamine in Southeast Asia, is diverted and smuggled out of China and India, whereas caffeine, the adulterant used for producing methamphetamine tablets, is mainly smuggled into Myanmar through its border with Thailand. Seizure data showed a dramatic increase in trafficking in MDMA through Southeast Asia. In terms of the drug epidemic, in 2002, cannabis remained overall the main drug of abuse in all of the countries of Southeast and Southwest Asia. Opiates, mainly opium and heroin, were also the drugs of choice except in Thailand, where opiate abuse declined, but ATS was the main drug of abuse due to its low cost and availability. A significant increase in ATS abuse, amphetamine, methamphetamine, and MDMA among the youth who smoked, sniffed, and inhaled them was reported in Cambodia, China, Indonesia, Laos, Myanmar, the Philippines, and Thailand. Injecting drug use among opiate abusers has been identified as the prime cause of the rapid spread of HIV/AIDS in Southeast and Southwest Asia. PMID- 15542749 TI - Recent trends of drug abuse and drug-associated deaths in Korea. AB - Methamphetamine is the most abused drug in Korea followed by cannabis and opiates. Recent characteristics of the drug problem in Korea include increased drug smuggling from abroad, drug trafficking by organized gangs, varieties of drug smuggling, foreigners engaged in drug smuggling, and spread among drug abusers and areas. New drugs such as MDMA, Yaba, and LSD are found in greater proportion in the seizure records, indicating diversification of smuggled drugs in Korea. In addition, there is a growing tendency for the abuse of common medicines among young people in Korea because they are easily available. Methamphetamine is so seriously abused that fatalities from its overdose have occurred; since 1985, 20 such fatalities have been reported. Many deaths from the abuse of noncontrolled substances, especially dextromethorphan, zipeprol, and carisoprodol, which are taken for their hallucinogenic effects, were also reported. Recently, there was even a fatality related to smuggling of cocaine by body-packing. PMID- 15542750 TI - Drug abuse trends among youth in the United States. AB - Several years of survey data indicate that illicit drug use among U.S. youth was at its highest level in 1979. Generally, the use of some illicit drugs declined progressively throughout the 1980s, stabilized, and then decreased slightly. Reducing illicit drug use among U.S. youth continues to be a priority of federal, state, and local law enforcement agencies. The U.S. government tracks youth illicit drug use through three nationally representative surveys: National Household Survey on Drug Abuse, Monitoring the Future Study, and Youth Risk Behavior Survey. Other surveys include the Partnership Attitudes Tracking Study, conducted annually to assess youth and parental attitudes about drugs, and Drug Abuse Awareness Network (DAWN), a national surveillance system that monitors trends in drug-related emergency department visits and deaths. Although survey methods differ, similar trends in drug use are well documented by these surveys. Among American youth, illicit use of drugs such as tobacco, alcohol, inhalants, marijuana, LSD, cocaine, heroin, and methamphetamine has remained relatively stable. However, a noteworthy exception is the increased use of MDMA (methylenedioxymethamphetamine). The physical consequences of "club and recreational drug use" are evidenced in the increased numbers of emergency department visits, specifically those related to MDMA and gamma-hydroxy-butyrate use, which may represent a new and emerging trend in illicit drug use. PMID- 15542751 TI - Influence of genetic variations of ethanol-metabolizing enzymes on phenotypes of alcohol-related disorders. AB - Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase-2 (ALDH2) play central roles in the metabolism of ethanol and its metabolite, acetaldehyde, in the liver. In ADH2, one nucleotide replacement causes either a super-active beta 2 subunit encoded by the ADH2*2 allele or a less active beta 1 subunit (ADH2*1 allele). In the same way, a G/A replacement at codon 487 of the ALDH2 gene produces an inactive form of the enzyme. Because the geno-types of these genes may explain individual differences in concentration and elimination of ethanol and acetaldehyde in the blood after drinking, they could be used as models to elucidate the contribution of these substances to the development of addiction and various types of organ damage. We have examined the influence of genetic variations of these enzymes on alcohol-related disorders in the Japanese. The results revealed that (1) the less active allele of the ADH2 gene (ADH2*1) is associated with an increased risk for alcohol dependence, alcohol-induced persistent amnestic disorder, alcohol withdrawal syndrome, and cancer of the upper GI tract; (2) the inactive allele of the ALDH2 gene (ALDH2*2) is associated with a decreased risk for alcohol dependence, and an increased risk for alcoholic polyneuropathy and cancer in the same region; and (3) these genetic variations modify clinical features of alcohol dependence. Possible mechanisms of altered risk for these disorders are discussed. PMID- 15542752 TI - Clinical features of nicotine dependence compared with those of alcohol, methamphetamine, and inhalant dependence. AB - A new clinical evaluation form was developed to compare the clinical features of nicotine dependence with those associated with other abused drugs. A new scoring system for clinical evaluation was developed. The form consisted of five scoring items: subjective effects, liking (of drug), withdrawal syndrome, acute psychic and physical disorders, and social disturbance. A preliminary clinical investigation was performed to test the validity of the evaluation form. Study subjects were those showing dependence on nicotine (cigarette smoking, n = 40), alcohol (n = 39), methamphetamine (n = 31), and inhalants (n = 30), who fulfilled the DSM-IV-TR criteria for drug dependence disregarding the state of "a maladaptive pattern of substance use, leading to clinically significant impairment or distress," and gave written informed consent for participation in the study. Nicotine caused a mild or the least degree of subjective effects, liking, and psychic and physical withdrawal symptoms, without any significant social disturbance or acute disorders. With alcohol, liking, withdrawal syndrome, and acute physical disorders were prominent. Methamphetamine produced the most serious acute psychic disorders, with intensive acute physical disorders and psychic withdrawal symptoms. Inhalants were characterized by an intensive degree of acute psychic disorders. As for social disturbance, alcohol, methamphetamine, and inhalants showed more significant influence than nicotine. Our study findings revealed that the clinical features of drug dependence could be evaluated by using the new clinical evaluation form. Further study is required to clarify the clinical features of nicotine dependence compared with those of other drugs of dependence. PMID- 15542753 TI - Influence of nicotine on brain reward systems: study of intracranial self stimulation. AB - We used the rate-frequency curve-shift procedure to evaluate the effects of nicotinic blockers, locally infused into the mesopontine tegmentum or ventral tegmentum, on the threshold of brain stimulation reward. Mecamylamine, the nicotinic acetylcholine receptor blocker, was infused one hour before the self stimulation of the medial forebrain bundle. When injected into the mesopontine tegmentum, mecamylamine shifted rate-frequency curves to the right. Similar effects were also observed when the drug was injected into the ventral tegmentum. Thus, in both the mesopontine tegmentum and the ventral tegmentum, nicotinic receptors appear to facilitate the rewarding effect of the self-stimulation. PMID- 15542754 TI - Role of gamma-aminobutyric acid (GABA) and metabotropic glutamate receptors in nicotine reinforcement: potential pharmacotherapies for smoking cessation. AB - Previous work indicated a role for GABA and glutamate in the reinforcing effects of drugs of abuse. The present studies assessed the effects of GABAergic and glutamatergic manipulations on the reinforcing effects of nicotine as assessed by intravenous nicotine self-administration. Male Wistar rats were allowed to self administer either of two nicotine doses under a fixed ratio or a progressive ratio schedule of reinforcement. The effects of a glutamatergic compound on nicotine self-administration in male DBA/2J mice were also explored. Finally, to assess for nonspecific effects of the drug manipulations, the effects of all test compounds on responding maintained by a food reinforcer were investigated. The pharmacological manipulations used were: gamma-vinyl-GABA (vigabatrin or GVG), an irreversible inhibitor of GABA transaminase, the GABAB receptor agonists ( )baclofen and CGP44532, and the metabotropic glutamate receptor 5 (mGluR5) antagonist MPEP. GVG, CGP44532, and (-)baclofen dose-dependently decreased nicotine self-administration on the fixed-ratio schedule, but also decreased food maintained responding. Furthermore, CGP44532 decreased breakpoints for nicotine and food at identical doses under the progressive-ratio schedule. MPEP dose dependently decreased nicotine self-administration with no effect on food maintained responding in rats. MPEP also decreased nicotine self-administration in the mice. These results demonstrate that activation of GABAB receptors or blockade of mGluR5 decreased nicotine self-administration. Although there was some selectivity for the effects of the GABAergic manipulations, there was clear selectivity of the effects of MPEP on nicotine- versus food-maintained responding. Thus, compounds that increase GABAergic neurotransmission and antagonists at mGluR5 have potential as anti-smoking medications for humans. PMID- 15542755 TI - Nicotine attenuates relapse to methamphetamine-seeking behavior (craving) in rats. AB - This study clarifies the modulating action of the nicotinic cholinergic system on reinstatement of methamphetamine (MAP)-seeking behavior (craving) using an intravenous, self-administration paradigm in rats. After self-administration of MAP for 10 days, replacing MAP with saline solution (MAP withdrawal) gradually decreased lever-pressing responses. On the sixth day of MAP withdrawal, MAP (1.0 mg/kg, i.p.)-priming injection significantly increased lever-pressing responses (reinstatement of MAP-seeking behavior). This MAP-seeking behavior was attenuated by repeated nicotine administration for 5 days during MAP withdrawal, and this attenuating effect was antagonized by the nicotinic antagonist mecamylamine. These results suggest that the appearance of MAP-seeking behavior may be due to inactivation of the nicotinic cholinergic neuron. Furthermore, it is suggested that nicotinic activating agents may be useful in preventing relapse to drug abuse. PMID- 15542756 TI - Alpha 7-type nicotinic acetylcholine receptor and prodynorphin mRNA expression after administration of (-)-nicotine and U-50,488H in beta-amyloid peptide (25 35)-treated mice. AB - We previously reported that (-)-nicotine and kappa-opioid receptor agonists lessened impairment of learning and/or memory in several animal models. Furthermore, these drugs prevented neurodegenerative damage induced by ischemia or beta-amyloid peptide (25-35). In the present study, we tested whether (-) nicotine and U-50,488H prevent delayed-memory impairment induced by beta-amyloid peptide (25-35), and changes of expression of alpha7-type nicotinic acetylcholine receptor mRNA and prodynorphin mRNA. Seven days after treatment with beta-amyloid peptide (25-35) (9 nmol/mouse, i.c.v.), memory impairment was observed in the Y maze test. Memory impairment was prevented when (-)-nicotine (6.16 micromol/kg, s.c.) or U-50,488H (21 micromol/kg, s.c.) was administered 1 h before, but not 1 h after, beta-amyloid peptide (25-35) treatment. There was no change in prodynorphin mRNA or alpha7-type nicotinic acetylcholine receptor mRNA expression in the hippocampus 10 days after beta-amyloid peptide (25-35) treatment alone. Of interest, mRNA expression of not only prodynorphin, but also the alpha7-type nicotinic acetylcholine receptor, was significantly decreased when U-50,488H was administered 1 h before, but not 1 h after, treatment with beta-amyloid peptide (25-35). However, these changes were not observed after the administration of (-) nicotine. These results suggest that activation of the kappa-opioid system, but not beta7-type nicotinic receptors has a neuroprotective effect on beta-amyloid peptide (25-35)-induced memory impairment, and may be involved in the long lasting changes in the expression of these mRNAs. PMID- 15542757 TI - Role of endocannabinoids and cannabinoid CB1 receptors in alcohol-related behaviors. AB - This review presents the remarkable research during the past several years indicating that some of the pharmacological and behavioral effects of alcohol, including alcohol drinking and alcohol-preferring behavior, are mediated through one of the most abundant neurochemical systems in the central nervous system, the endocannabinoid signaling system. The advances, with the discovery of specific receptors and the existence of naturally occurring cannabis-like substances in the mammalian system and brain, have helped in understanding the neurobiological basis for drugs of abuse, including alcoholism. The cDNA and genomic sequences encoding G-protein-coupled cannabinoid receptors (CB1 and CB2) from several species have now been cloned. This has facilitated discoveries of endogenous ligands (endocannabinoids). To date, two fatty acid derivatives characterized to be arachidonylethanolamide and 2-arachidonylglycerol have been isolated from both nervous and peripheral tissues. Both these compounds have been shown to mimic the pharmacological and behavioral effects of Delta9-tetrahydrocannabinol, the psychoactive component of marijuana. The involvement of the endocannabinoid signaling system in tolerance development to drugs of abuse, including alcohol, were unknown until recently. Studies from our laboratory demonstrated for the first time the downregulation of CB1 receptor function and its signal transduction by chronic alcohol. The observed downregulation of CB1 receptor binding and its signal transduction results from the persistent stimulation of receptors by the endogenous CB1 receptor agonists arachidonylethanolamide and 2 arachidonylglycerol, the synthesis of which is increased by chronic alcohol treatment. The deletion of CB1 receptor has recently been shown to block voluntary alcohol intake in mice, which is consistent with our previous findings where the DBA/2 mice known to avoid alcohol intake had significantly reduced brain CB1 receptor function. These findings suggest a role for the CB1 receptor gene in excessive alcohol drinking behavior and development of alcoholism. Ongoing investigations may lead to the development of potential therapeutic agents to modulate the endocannabinoid signaling system, which will be helpful for the treatment of alcoholism. PMID- 15542758 TI - 4-methylpyrazole decreases 1,4-butanediol toxicity by blocking its in vivo biotransformation to gamma-hydroxybutyric acid. AB - 1,4-Butanediol (1,4-BD), a prodrug converted in vivo to gamma-hydroxybutyric acid by alcohol dehydrogenase, has resulted in life-threatening overdoses and deaths. We investigated whether 4-methylpyrazole (4-MP), an alcohol dehydrogenase antagonist, can be used as an antidote in a murine model of 1,4-BD overdose. CD-1 mice were overdosed with 1,4-BD, 600 mg/kg i.p. Mice then received 4-MP, 25 mg/kg i.p., or control injections after 1 min, 5 min, and symptom appearance. Mice were then evaluated for toxicity by the righting reflex and rotarod test every 10 min after intervention. When 4-MP was administered 1 and 5 min after 1,4-BD overdose, mice completely maintained their righting reflex. Conversely, control mice lost their righting reflex for 110 and 130 min, respectively (P < 0.05). When 4-MP was administered after symptomatic 1,4-BD overdose, mice lost their righting reflex but recovered it by 60 min. Conversely, control mice lost their righting reflex and recovered it by 140 min (P < 0.05). When 4-MP was administered at 1 min after 1,4-BD overdose, mice never failed the rotarod test. Conversely, control mice failed the rotarod test for 210 min (P < 0.05). When 4-MP was administered 5 min after 1,4-BD and after symptomatic 1,4-BD overdose, mice failed the rotarod test for 100 and 110 min, respectively. Conversely, control mice failed the rotarod test for 210 and 180 min, respectively (P < 0.05). In addition, treatment of mice with 4-MP significantly attenuated increases in blood gamma-hydroxybutyric acid concentrations and prevented loss of the righting reflex and failure of the rotarod test. In this murine model of 1,4-BD overdose, 4-MP conferred antidotal effects by inhibiting alcohol dehydrogenase-mediated biotransformation of 1,4-BD to gamma-hydroxybutyric acid. PMID- 15542759 TI - Evaluation of gamma-hydroxybutyric acid for genotoxicity in the mouse micronucleus assay. AB - Gamma-hydroxybutyric acid (GHB) is an endogenous compound found in the brain and other tissues of mammals. Neurotransmitter/neuromodulator functions have been ascribed to GHB, which has lately become a drug of abuse. In this study, we tested GHB for genotoxicity by measuring its ability to induce micronuclei in polychromatic erythrocytes (reticulocytes) in the peripheral blood of mice. Intraperitoneal injection with a dose of 25 mg/kg/day for 3 days or 50 mg/kg/day x 3 days resulted in a significant (by Dunnett's test) increase of 1.9- to 2.1 fold in micronuclei. However, because increases were small and because no consistent dose-dependent increase in induced micronuclear frequency could be demonstrated, our results do not conclusively show that GHB is an in vivo genotoxicant in mammals. PMID- 15542760 TI - Repeated exposure to the abused inhalant toluene alters levels of neurotransmitters and generates peroxynitrite in nigrostriatal and mesolimbic nuclei in rat. AB - Toluene, a volatile hydrocarbon found in a variety of chemical compounds, is misused and abused by inhalation for its euphorigenic effects. Toluene's reinforcing properties may share a common characteristic with other drugs of abuse, namely, activation of the mesolimbic dopamine system. Prior studies in our laboratory found that acutely inhaled toluene activated midbrain dopamine neurons in the rat. Moreover, single systemic injections of toluene in rats produced a dose-dependent increase in locomotor activity which was blocked by depletion of nucleus accumbens dopamine or by pretreatment with a D2 dopamine receptor antagonist. Here we examined the effects of seven daily intraperitoneal injections of 600 mg/kg toluene on the content of serotonin and dopamine in the caudate nucleus (CN) and nucleus accumbens (NAC), substantia nigra, and ventral tegmental area at 2, 4, and 24 h after the last injection. Also, the roles of nitric oxide, peroxynitrite, and the production of 3-nitrosotyrosine (3-NT), in the CN and NAC were assessed at the same time points. Toluene treatments increased dopamine levels in the CN and NAC, and serotonin levels in CN, NAC, and ventral tegmental area. Measurements of the dopamine metabolite dihydroxyphenylacetic acid (DOPAC) further suggested a change in transmitter utilization in CN and NAC. Lastly, 3-NT levels also showed a differential change between CN and NAC, but at different time points post-toluene injection. These results point out the complexity of action of toluene on neurotransmitter function following a course of chronic exposure. Changes in the production of 3 NT also suggest that toluene-induced neurotoxicity may mediate via generation of peroxynitrite. PMID- 15542761 TI - The role of N-methyl-D-aspartate receptors in neurobehavioral changes induced by toluene exposure during synaptogenesis. AB - Toluene abuse during pregnancy results in newborns with fetal solvent syndrome. N Methyl-D-aspartate (NMDA) receptor has been identified as a target site for toluene. Since the normal function of NMDA receptor is critical for synaptogenesis, the long-term effects of toluene exposure during synaptogenesis on the neurobehavioral function and the expression of NMDA receptor subunits (NR1, NR2A, and NR2B) were examined. Rats exposed to l g/kg of toluene (i.p.) over postnatal days 4 to 9 were found to exhibit reduction in body weight, NMDA induced seizure thresholds, and MK-801-induced hyperlocomotor activity. Furthermore, immunoblotting and immunohistochemical analysis revealed a significant increase in NR2A subunit expression in the hippocampus and cerebellum of toluene-exposed rats on PN30. These results suggest that the region-specific changes in the expression of NMDA receptor subunits may play a role in the neurobehavioral dysfunction following toluene exposure during synaptogenesis. PMID- 15542762 TI - Neonatal toluene exposure alters glutamate-induced calcium signaling in developing cerebellar granule neurons. AB - Glutamatergic neurotransmission is critical both for neurogenesis and mature functioning of the central nervous system (CNS), and is thought to be one target for toluene-induced damage. It has been reported that toluene antagonizes the function of N-methyl-D-aspartate (NMDA) receptor. In this study, we examined the effect that neonatal toluene exposure has on NMDA receptor in cerebellar granule neurons. Sprague-Dawley rats were treated with 0, 200, 500, and 1000 mg/kg of toluene by intraperitoneal injection from postnatal day (PN) 4 to 7. After culture under toluene-free condition, Ca2+ signaling in response to glutamate and NMDA was measured using fura-2 Ca2+ imaging for up to 14 days. Toluene exposure dose-dependently reduced glutamate/glycine and NMDA/glycine induced Ca2+ signaling in culture cerebellar granule neurons at DIV5 (day in vitro), and the effects were gradually recovered. The effects of toluene exposure on NMDA stimulated Ca2+ signals in response to NMDA receptor inhibitors were also compared. The results indicated that neonatal toluene exposure can induce long term but reversible changes in NMDA-induced Ca2+ signaling pathway. Such changes could be involved in the impairment of CNS function and development observed in fetal solvent syndrome. PMID- 15542763 TI - Using a DNA microarray method to examine gene expression in brain from clozapine injected mice. AB - After our 2002 report on the changes of gene expression in the brain from phencyclidine-treated mouse by using DNA microarray (Toyooka et al., Ann. N.Y. Acad. Sci. 965: 10-20), we decided to apply this DNA microarray method for the brain of the mouse treated with other drugs. We are now examining the effects of clozapine on the brain function. Clozapine is an atypical antipsychotic drug. Clozapine has affinity for neurotransmitter receptors, including D4 dopamine, serotonin, histamine, and adrenergic receptors. MacGibbon et al. (Mol. Brain Res. 23: 21-32) reported that clozapine and haloperidol produced a differential pattern of immediate early gene expression in rat caudate-putamen. Kobayashi et al. (Br. J. Pharmacol. 123: 421-426) have observed the effects of clozapine on the opioid receptors and G-protein-activated inwardly rectifying potassium channel of Xenopus oocytes. Thomas et al. (J. Neurochem. 76: 789-796) found that clozapine increases apolipoprotein D expression in the mouse brain. Therefore, we are going to apply this DNA microarray method to examine the effects of clozapine on mouse brain function. After we injected clozapine into mice for 20 days, we decapitated the mice. We then used the DNA microarray method to examine the gene expression of mouse brain. We found some changes in the mouse brain treated with clozapine. For example, the intensity of a potassium channel spot decreased, and that of a serotonin receptor spot increased. PMID- 15542764 TI - Comparison of G-protein selectivity of human 5-HT2C and 5-HT1A receptors. AB - We compared the ability of human 5-HT2C and 5-HT1A receptors to couple to selected G proteins expressed in insect Sf9 cells through simultaneous infection with recombinant baculoviruses. We also examined the coupling of G proteins to these same receptors in membranes derived from the Sf9 cells using in situ reconstitution with purified G proteins. Our data show that unoccupied 5-HT2C and 5-HT1A receptors can attain an activated conformation that is stabilized by interaction with specific G proteins. While high-affinity agonist binding to the 5-HT2C receptor was increased to a greater extent by Galphaq than by Galphai2, the high-affinity agonist binding to the 5-HT1A receptor was preferentially enhanced by Galphai2 coexpression. When the two 5-HT receptors were expressed in cells also expressing G proteins, both 5-HT2C and 5-HT1A receptors appear to activate Galphai2 in preference to Galphaq. In contrast, in situ reconstitution data show that 5-HT2C receptors robustly activate Galphaq and marginally activate Galphao or Galphai, whereas 5-HT1A receptors only marginally activate Galphaq and robustly activate Galphao and Galphai. These results suggest that the overexpression of receptor and potential G-protein coupling partners in Sf9 cells may lead to erroneous conclusions as to the signaling selectivity of receptors. PMID- 15542765 TI - Pharmacological basis for management of drug dependence. AB - Neurochemical mechanisms underlying development of drug dependence and withdrawal syndrome remain unclear. Several clinical features of withdrawal syndrome, such as anxiety, are considered to be common among patients with drug dependence induced by different drugs of abuse. In the present study, we investigated whether diazepam-binding inhibitor (DBI), an endogenous anxiogenic neuropeptide, participates in the anxiety associated with drug dependence and its withdrawal symptoms. When we examined brain from mice dependent on alcohol, nicotine, and morphine, we observed that the levels of DBI protein and its mRNA significantly increased. Abrupt cessation of these drugs facilitated further increases in DBI expression. In the cases of nicotine- and morphine-dependent mice, concomitant administration of specific antagonists for nicotinic acetylcholine and m-opioid receptors, respectively, abolished the increased expression. Similar patterns of DBI expression were observed in the neurons after sustained exposure to these drugs and its removal from culture medium. Sustained exposure of the neurons to abused drugs significantly increased the KCl (30 mM)-induced 45Ca2+ influx and enhanced expression of alpha1 subunits for L-type high voltage-gated Ca2+ channels (HVCCs). In addition, the increase in DBI expression was completely blocked by L-type HVCC inhibitors. Therefore, these alterations in DBI expression, mediated via increased influx of Ca2+ through upregulated L-type HVCCs, are closely related to drug dependence and/or its withdrawal syndrome and are considered to be part of a common biochemical process in drug dependence induced by different drugs of abuse. PMID- 15542766 TI - Monitoring drug-induced neurodegeneration by imaging of peripheral benzodiazepine receptors. AB - Several drugs of abuse are known to produce an array of deleterious effects, including alterations in neuronal circuitry and, ultimately, neuronal degeneration. For instance, methamphetamine was shown to induce substantial nigrostriatal dopaminergic terminal damage, including an increase in glial fibrillary acidic protein, a marker for astrocyte proliferation. Nevertheless, there was almost no attempt to define neurodegeneration by measuring the abundance of reactive microglia. In fact, some investigators fail to differentiate between astrocytes and microglia and claim glial fibrillary acidic protein to be a marker for gliosis. To date, there are numerous methods designed to assess brain neuropathologies resulting from a wide arsenal of insults. Regardless of the cause of neuronal damage, reactive glial cells always appear at and around the site of degeneration. These cells are distinguished by the exceptional abundance of peripheral benzodiazepine receptors (PBRs; omicron3 sites), particularly as compared to surrounding neurons. Measuring the binding of specific ligands to these PBRs (for example, [3H]PK 11195) offers a unique indirect marker for reliable impairment estimation in the central nervous system. Moreover, the availability of agents such as [11C]PK 11195 paved the road to in vivo animal and human brain positron emission tomography scanning, demonstrating inflammation-like processes in several diseases. Additionally, the measurement of increased binding of PBR ligands provides a faithful indicator for the behavioral and cognitive deficits accompanying neuronal injury. PMID- 15542767 TI - Modulators of G protein-activated inwardly rectifying K+ channels: potentially therapeutic agents for addictive drug users. AB - G protein-activated inwardly rectifying K+ (GIRK, Kir3) channels play an important role in the inhibitory regulation of neuronal excitability in most brain regions and heart rate through activation of various G protein-coupled receptors, such as opioid, cannabinoid, and D2 dopamine receptors. Therefore, modulators of GIRK channels may affect many brain functions. We have shown using Xenopus oocyte expression assays that ethanol directly activates GIRK channels, whereas various antipsychotics (thioridazine, clozapine, pimozide, and haloperidol) inhibit the channels. Here we investigated not only the effects of various selective serotonin reuptake inhibitor (SSRI) antidepressants (fluoxetine, citalopram, fluvoxamine, and zimelidine) and risperidone, an atypical antipsychotic, on GIRK channels, but also those of the various drugs tested on other Kir channels using the Xenopus oocyte system. Fluoxetine inhibited GIRK channels, whereas the other SSRIs and risperidone had a small or no effect on the channels. In contrast, Kir1.1 and Kir2.1 channels were insensitive to ethanol and various SSRIs and antipsychotics, although thioridazine weakly inhibited Kir1.1 channels. It has been shown that the function of GIRK channels is involved in seizure susceptibility, antinociception by opioids, cannabinoids, or ethanol, and cocaine reinforcement in studies using GIRK knockout mice and weaver mutant mice that have mutant GIRK2 channels insensitive to G proteins and ethanol. Activation of GIRK channels by opioids, cannabinoids, or ethanol may be one of these key effects. Therefore, GIRK channel modulators might be potential agents for the treatment of users of addictive drugs, such as cocaine, opioids, cannabinoids, and ethanol, as well as for the treatment of epilepsy and pain. PMID- 15542768 TI - Convergent effects on cell signaling mechanisms mediate the actions of different neurobehavioral teratogens: alterations in cholinergic regulation of protein kinase C in chick and avian models. AB - Although the actions of heroin on central nervous system (CNS) development are mediated through opioid receptors, the net effects converge on dysfunction of cholinergic systems. We explored the mechanisms underlying neurobehavioral deficits in mouse and avian (chick, Cayuga duck) models. In mice, prenatal heroin exposure (10 mg/kg on gestation days 9-18) elicited deficits in behaviors related to hippocampal cholinergic innervation, characterized by concomitant pre- and postsynaptic hyperactivity, but ending in a reduction of basal levels of protein kinase C (PKC) isoforms betaII and gamma and their desensitization to cholinergic receptor-induced activation. PKCalpha, which is not involved in the behaviors studied, was unaffected. Because mammalian models possess inherent confounding factors from maternal effects, we conducted parallel studies using avian embryos, evaluating hyperstriatal nucleus (intermedial part of the hyperstriatum ventrale, IMHV)-related, filial imprinting behavior. Heroin injection to the eggs (20 mg/kg) on incubation days 0 and 5 diminished the post-hatch imprinting ability and reduced PKCg and bII content in the IMHV membrane fraction. Two otherwise unrelated agents that converge on cholinergic systems, chlorpyrifos and nicotine, elicited the same spectrum of effects on PKC isoforms and imprinting but had more robust actions. Pharmacological characterization also excluded direct effects of opioid receptors on the expression of imprinting; instead, it indicated participation of serotonergic innervation. The avian models can provide rapid screening of neuroteratogens, exploration of common mechanisms of behavioral disruption, and the potential design of therapies to reverse neurobehavioral deficits. PMID- 15542769 TI - Effects of neonatal exposure to methamphetamine: catecholamine levels in brain areas of the developing rat. AB - Neonatal exposure to moderate doses of methamphetamine during the first month of life in the rat affects tyrosine hydroxylase gene expression in the substantia nigra and nigrostriatal tyrosine hydroxylase activity. The main goal of this work was to evaluate the ontogeny of the neurochemical effects of repeated exposure to moderate doses of methamphetamine during the first month of life in the rat. Norepinephrine, dopamine, and dihydroxyphenylacetic acid levels were measured in target areas of methamphetamine: the substantia nigra, ventral tegmental area, caudate-putamen, nucleus accumbens, and medial prefrontal cortex. On postnatal day 1 (PND1), Wistar rat litters, culled to eight pups, sex balanced, were randomly attributed to either methamphetamine or control groups. Methamphetamine groups were administered 10 mg of (+/-)-methamphetamine/kg body weight/day, subcutaneously, from PND1 until the day prior to sacrifice; control groups received isovolumetric saline. Groups were sacrificed on PND7, PND14, and PND30. Neonatal methamphetamine exposure increased norepinephrine levels in the substantia nigra of PND30 rats; on PND14, this variation was evident only in male pups. In the substantia nigra, the dihydroxyphenylacetic/dopamine ratio was also affected in PND30 males. In the ventral tegmental area, catecholamine levels were not affected by methamphetamine. Norepinephrine levels were also increased in the caudate-putamen of PND7 male and PND14 female methamphetamine-exposed pups and in the nucleus accumbens of PND14 female and PND30 male and female pups. Catecholamine levels in the medial prefrontal cortex were not affected by neonatal methamphetamine administration. PMID- 15542770 TI - Conditioned place preference and locomotor sensitization after repeated administration of cocaine or methamphetamine in rats treated with epidermal growth factor during the neonatal period. AB - Epidermal growth factor (EGF) and its structurally related proteins are involved in the developmental regulation of various brain neurons, including midbrain dopaminergic neurons. We recently reported EGF and EGF-receptor abnormalities in both the brain tissues and blood of schizophrenic patients. Administration of EGF to neonatal rats transiently increases tyrosine hydroxylase expression and subsequently results in behavioral abnormalities in prepulse inhibition of acoustic startle, locomotor activity, and social interaction after development. The enhanced locomotor and stereotypic responses of the neonatally EGF-treated rats are considered to be an animal model for positive schizophrenia symptoms. In the present study, we investigated psychostimulant sensitivity of neonatally EGF treated rats. At the adult stage, EGF-treated rats were challenged with cocaine (15 mg/kg) or methamphetamine (1 mg/kg), and conditioned place preference and locomotor activity were examined. The rats that received EGF during the neonatal period had significantly higher conditioned place preference for where cocaine or methamphetamine was administered than controls. The neonatal EGF treatment enhanced behavioral response to methamphetamine and behavioral sensitization to cocaine at the adult stage. Drug-naive controls gradually increased locomotor responses to cocaine during their daily injections, whereas EGF-treated rats exhibited a larger increase in cocaine responses. These results indicate that overactivation of the EGF receptors (ErbB1) during the neonatal period influences future sensitivity to psychostimulants. Our findings indicate a potential link between EGF-receptor activation and drug addiction. PMID- 15542771 TI - Effects of postnatal cocaine exposure and environmental enrichment on rat behavior in a forced swim test. AB - This study examined the effects of environmental enrichment on rats exposed to cocaine during the first month of life, in several categories of behavior observed in a forced swim test. Wistar rats were divided in four groups. The first included pups that were subjected to injections of cocaine hydrochloride (15 mg/kg body weight/day, subcutaneously, in two daily doses, from postnatal days 1 to 27) and reared in an enriched environment (CocEE); the second, pups that were subjected to injections of cocaine (as previously described) and reared in a standard environment (CocSE); the third, pups that were subjected to saline injections and reared in an enriched environment (SalEE); the fourth, pups that were subjected to saline injections and reared in a standard environment (SalSE). On postnatal days 26 and 27, rats were tested in a swimming pool in two 5-min sessions. The categories of behavior studied in this work were: fast swim, slow swim, struggling, diving, and immobility. Results showed that postnatal cocaine exposure decreased the time spent on fast swim during the two sessions and increased the immobility behavior during the second session in CocSE pups compared with SalSE pups. SalEE pups increased the time spent in fast swim, slow swim, and diving, and decreased the time spent in struggling and immobility during the two sessions compared with SalSE pups. CocEE animals spent more time in fast swim and struggling and less the time in immobility compared with CocSE pups. The present results suggest that postnatal cocaine exposure affects the ability of these animals to cope with stressful situations, and that environmental enrichment seems to enable the rats to adopt a more active strategy, one that allows them to better cope with this particular stress situation. PMID- 15542772 TI - Abnormal immunoreactivity to serotonin in cerebellar Purkinje cells after neonatal cocaine exposure. AB - Neonatal cocaine is known to affect the developing serotonergic system in many brain structures, including the cerebellum. Changes in the cerebellar Purkinje cells after drug exposure are well documented and result in impairment of movement and other cerebellar disorders such as ataxia. These cells have a major postnatal developmental pattern; therefore, neonatal exposure to cocaine is likely to affect them. In this work, male and female Wistar rats were injected with 15 mg of cocaine hydrochloride/kg body weight/day, subcutaneously, in two daily doses, from postnatal day 1 (PND1) to PND29. Controls were given 0.9% of saline. On PND14, PND21, and PND30, rats were transcardially perfused, and brains removed and cryoprotected. Coronal sections from the cerebellum were processed for immunocytochemistry of cells containing serotonin (5-hydroxytryptamine, or 5 HT). At the same postnatal age, rats from at least three different litters were sacrificed by decapitation, and brains were dissected for determination of 5-HT in the cerebellum by high-performance liquid chromatography with electrochemical detection. Upon the expected distribution of immunoreactivity to 5-HT, an abnormal immunoreactivity to 5-HT was observed in the Purkinje cells of six cocaine-exposed animals, but not in control animals. Also, levels of cerebellar 5 HT in cocaine-exposed rats were significantly increased on PND21. These results, together with previously reported observations of altered patterns of motor behavior, indicate that neonatal cocaine exposure affects the serotonergic cerebellar system, altering the standard development of Purkinje cells and possibly compromising the motor function. PMID- 15542773 TI - Sequence and helicity requirements for the proapoptotic activity of Bax BH3 peptides. AB - Overexpression of the antiapoptotic proteins Bcl-2 and Bcl-XL is commonly observed in human malignancies and contributes to chemotherapy and radiation resistance. Bcl-2 and Bcl-XL inhibit apoptosis by binding to proapoptotic proteins such as Bax, thereby preventing chemotherapy-induced or radiation induced release of cytochrome c from mitochondria and subsequent activation of the caspase protease cascade. Efforts to inhibit Bcl-2 or Bcl-XL function in tumor cells have focused on developing agents to inhibit the interactions of these proteins with proapoptotic proteins. Peptides derived from the BH3 domains of proapoptotic proteins have been shown to disrupt the interactions of Bcl-2 and Bcl-XL with key binding partners in cell-free reactions and to promote cellular apoptosis. However, less is known about the targets of BH3 peptides in intact cells as well as the sequence, length, and conformational requirements for peptide biological activity. In this report, we show that cell-permeable Bax BH3 peptides physically disrupt Bax/Bcl-2 heterodimerization in intact cells and that this disruption correlates with peptide-induced cell death. A point-mutant, control peptide that failed to disrupt intracellular Bax/Bcl-2 interactions also failed to promote apoptosis. To determine important sequence, length, and structural requirements for peptide activity, we generated and systematically analyzed the biological activities of 17 Bax BH3 peptide variants. Peptides were quantitatively examined for their ability to inhibit Bax/Bcl-2 and Bax/Bcl-XL heterodimerization in vitro and to promote cytochrome c release from mitochondria isolated from Jurkat, HL-60, U937, and PC-3 cells. Our results define 15 amino acids as the minimal length required for Bax BH3 peptide biological activity and show that amino acids COOH terminal to the BH3 core sequence are less critical than those located NH2 terminal to the core. In addition, circular dichroism spectroscopy revealed that high alpha-helical content generally correlated with, but was not sufficient for, peptide activity. Taken together, these studies provide a basis for future optimization of Bax BH3 peptide as a therapeutic anticancer agent. PMID- 15542774 TI - Inhibitory effect of epidermal growth factor on resveratrol-induced apoptosis in prostate cancer cells is mediated by protein kinase C-alpha. AB - Resveratrol, a naturally occurring stilbene with antitumor properties, caused mitogen-activated protein kinase [MAPK, extracellular signal-regulated kinase 1/2 (ERK1/2)] activation, nuclear translocation of Ser15-phosphorylated p53, and p53 dependent apoptosis in hormone-insensitive DU145 prostate cancer cells. Exposure of these cells to epidermal growth factor (EGF) for up to 4 hours resulted in brief activation of MAPK followed by inhibition of resveratrol-induced signal transduction, p53 phosphorylation, and apoptosis. Resveratrol stimulated c-fos and c-jun expression in DU145 cells, an effect also suppressed by EGF. An inhibitor of protein kinase C (PKC)-alpha, -beta, and -gamma (CGP41251) enhanced Ser15 phosphorylation of p53 by resveratrol in the absence of EGF and blocked EGF inhibition of the resveratrol effect. EGF caused PKC-alpha/beta phosphorylation in DU145 cells, an effect reversed by CGP41251. Activation of PKC by phorbol ester (phorbol 12-myristate 13-acetate) enhanced EGF action on ERK1/2 phosphorylation without significantly altering p53 phosphorylation by resveratrol. DU145 cells transfected with a dominant-negative PKC-alpha construct showed resveratrol-induced ERK1/2 phosphorylation and Ser15 phosphorylation of p53 but were unresponsive to EGF. Thus, resveratrol and EGF activate MAPK by discrete mechanisms in DU145 cells. The stilbene promoted p53-dependent apoptosis, whereas EGF opposed induction of apoptosis by resveratrol via a PKC alpha-mediated mechanism. Resveratrol also induced p53 phosphorylation in LNCaP prostate cancer cells, an effect also inhibited by EGF. Inhibition of PKC activation in LNCaP cells, however, resulted in a reduction, rather than increase, in p53 activation and apoptosis, suggesting that resveratrol-induced apoptosis in these two cell lines occurs through different PKC-mediated and MAPK dependent pathways. PMID- 15542775 TI - Discovery and mechanism of action of a novel series of apoptosis inducers with potential vascular targeting activity. AB - A novel series of 2-amino-4-(3-bromo-4,5-dimethoxy-phenyl)-3-cyano-4H-chromenes was identified as apoptosis-inducing agents through our cell-based apoptosis screening assay. Several analogues from this series, MX-58151, MX-58276, MX 76747, MX-116214, MX-126303, and MX-116407, were synthesized and further characterized. MX-116407, a lead compound from this series, induced apoptosis with an EC50 of 50 nmol/L and inhibited cell growth with a GI50 of 37 nmol/L in T47D breast cancer cells. Treatment of cells with these analogues led to G2-M arrest, cleavage of essential proapoptotic caspase substrates, and induction of nuclear fragmentation. We identified these compounds as tubulin destabilizers with binding site at or close to the colchicine binding site. Compounds in this series were also active in drug-resistant cancer cell lines with a GI50 value for one of the analogues (MX-58151) of 2.5 nmol/L in paclitaxel-resistant, multidrug resistant MES-SA/DX5 tumor cells. This series of compounds displayed high selectivity against proliferating versus resting cells. Interestingly, these compounds were shown to disrupt preformed endothelial cell capillary tubules in vitro and affect functional vasculature to induce tumor necrosis in vivo and are thus likely to work as tumor vasculature targeting agents. Among these compounds, MX-116407 showed capillary tubule disruption activity in vitro at concentrations well below the cytotoxic dose. In a separate study, we further characterized the antitumor efficacy and pharmacokinetic profile of this series of compounds and identified MX-116407 as a potent apoptosis-inducing agent with apparent activity as tumor vasculature targeting agent. PMID- 15542776 TI - Antivascular and antitumor evaluation of 2-amino-4-(3-bromo-4,5-dimethoxy-phenyl) 3-cyano-4H-chromenes, a novel series of anticancer agents. AB - A novel series of 2-amino-4-(3-bromo-4,5-dimethoxy-phenyl)-3-cyano-4H-chromenes was identified as potent apoptosis inducers through a cell-based high throughput screening assay. Six compounds from this series, MX-58151, MX-58276, MX-76747, MX 116214, MX-116407, and MX-126303, were further profiled and shown to have potent in vitro cytotoxic activity toward proliferating cells only and to interact with tubulin at the colchicine-binding site, thereby inhibiting tubulin polymerization and leading to cell cycle arrest and apoptosis. Furthermore, these compounds were shown to disrupt newly formed capillary tubes in vitro at low nanomolar concentrations. These data suggested that the compounds might have vascular targeting activity. In this study, we have evaluated the ability of these compounds to disrupt tumor vasculature and to induce tumor necrosis. We investigated the pharmacokinetic and toxicity profiles of all six compounds and examined their ability to induce tumor necrosis. We next examined the antitumor efficacy of a subset of compounds in three different human solid tumor xenografts. In the human lung tumor xenograft (Calu-6), MX-116407 was highly active, producing tumor regressions in all 10 animals. Moreover, MX-116407 significantly enhanced the antitumor activity of cisplatin, resulting in 40% tumor-free animals at time of sacrifice. Our results identify MX-116407 as the lead candidate and strongly support its continued development as a novel anticancer agent for human use. PMID- 15542777 TI - Caspase-mediated apoptosis and caspase-independent cell death induced by irofulven in prostate cancer cells. AB - Irofulven (hydroxymethylacylfulvene) is a novel antitumor drug, which acts by alkylating cellular macromolecular targets. The drug is a potent inducer of apoptosis in various types of tumor cells, whereas it is nonapoptotic in normal cells. This study defined molecular responses to irofulven involving mitochondrial dysfunction and leading to death of prostate tumor LNCaP-Pro5 cells. Irofulven caused early (2-5 hours) translocation of the proapoptotic Bax from cytosol to mitochondria followed by the dissipation of mitochondrial membrane potential and cytochrome c release at 4 to 12 hours. These effects preceded caspase activation and during the first 6 hours were not affected by caspase inhibitors. Processing of caspase-9 initiated the caspase cascade at approximately 6 hours and progressed over time. The activation of the caspase cascade provided a positive feedback loop that enhanced Bcl-2-independent translocation and cytochrome c release. General and specific caspase inhibitors abrogated irofulven-induced apoptotic DNA fragmentation with the following order of potency: pan-caspase > or = caspase-9 > caspase-8/6 > caspase-2 > caspase-3/7 > caspase-1/4. Abrogation of caspase-mediated DNA fragmentation failed to salvage irofulven-treated cells from growth inhibition and loss of viability, demonstrating a substantial contribution of a caspase-independent cell death. Monobromobimane, an inhibitor of alternative caspase-independent apoptotic pathway that is mediated by mitochondrial permeability transition, antagonized both apoptosis, measured as phosphatidylserine externalization, and cytotoxicity of irofulven. Collectively, the results indicate that irofulven-induced signaling is integrated at the level of mitochondrial dysfunction. The induction of both caspase-dependent and caspase-independent death pathways is consistent with pleiotropic effects of irofulven, which include targeting of cellular DNA and proteins. PMID- 15542778 TI - The histone deacetylase inhibitor FR901228 induces caspase-dependent apoptosis via the mitochondrial pathway in small cell lung cancer cells. AB - Histone deacetylase inhibitors modulate the transcription of target genes and represent a new class of anticancer agents. The histone deacetylase inhibitor FR901228 has been reported to show antiproliferative and apoptotic effects in various malignancies including small cell lung cancer (SCLC) in vitro; however, the underlying mechanism is not fully understood. BCL-2 and BCL-XL are antiapoptotic proteins, of which overexpression has been reported to confer resistance to anticancer agents. High levels of BCL-2 and BCL-XL are frequently expressed in SCLC tumors. The present study was designed to clarify the apoptotic pathway of FR901228 in SCLC cells in vitro. FR901228 induced apoptosis in three SCLC cell lines after 24 hours of treatment. FR901228 activated caspase-9 and caspase-3 but not caspase-8, and the caspase-3 inhibitor Z-DEVD-fmk blocked the cytotoxicity of FR901228. FR901228 down-regulated the expression of bcl-2 and bcl xL mRNA through de novo protein synthesis and suppressed the expression of BCL-2 and BCL-XL proteins. In addition, the combination of bcl-2 antisense oligonucleotides with FR901228 enhanced FR901228-induced caspase-3 activity and cytotoxicity. These findings suggest that FR901228 induces caspase-dependent apoptosis via the mitochondrial pathway rather than the death receptor pathway. Considering the possible contributions of BCL-2 and BCL-XL to multidrug resistance, FR901228 is a promising agent in the treatment of refractory as well as primary SCLC tumors. PMID- 15542779 TI - Enhanced topoisomerase II targeting by annamycin and related 4-demethoxy anthracycline analogues. AB - Targeting topoisomerase II (topo II) is regarded as an important component of the pleiotropic mechanism of action of anthracycline drugs. Here, we show that 4 demethoxy analogues of doxorubicin, including annamycin, exhibit a greater ability to trap topo II cleavage complexes than doxorubicin and some other 4 methoxy analogues. In leukemic CEM cells with wild-type topo II, annamycin induced substantial levels of topo II-mediated DNA-protein cross-links (15-37% of total DNA for 0.5-50 micromol/L drug), whereas doxorubicin-induced DNA-protein cross-links were marginal (0-4%). In CEM/VM-1 cells that harbor mutated, drug resistant topo II, both 4-methoxy and 4-demethoxy drugs produced marginal DNA protein cross-links. Annamycin, but not doxorubicin, formed topo II-mediated DNA protein cross-links also in isolated CEM nuclei. In disparity with the unequal DNA-protein cross-link induction, both drugs induced comparable levels of DNA strand breaks in CEM cells. Compared with CEM, drug cytotoxicity against CEM/VM-1 cells was reduced 10.5- to 13.8-fold for 4-demethoxy analogues but only 3.8- to 5.5-fold for 4-methoxy drugs. Hence, growth inhibition by 4-demethoxy analogues seems more dependent on the presence of wild-type topo II. The enhanced topo II targeting by 4-demethoxy analogues was accompanied by a profound induction of apoptotic DNA fragmentation in leukemic CEM cells. Normal WI-38 fibroblasts, however, were markedly more resistant to annamycin-induced DNA-protein cross links, apoptosis, and growth inhibition. The enhanced topo II targeting by 4 demethoxy doxorubicin analogues underscores the mechanistic diversity of anthracycline drugs. This diversity needs to be recognized as a factor in responses to drugs such as annamycin and doxorubicin. PMID- 15542780 TI - 8-amino-adenosine is a potential therapeutic agent for multiple myeloma. AB - Multiple myeloma (MM) is a malignancy of clonal B-cells that accounts for 10% of all hematologic malignancies. We have shown previously that a novel purine analogue, 8-chloro-adenosine, has significant activity for MM in preclinical studies. OBJECTIVE: Using MM cell lines, we investigated the molecular mechanism of related congener of adenosine, 8-amino-adenosine (8-NH2-Ado). METHODS: We employed biological and biochemical assays in MM cell lines to evaluate the clinical potential of 8-NH2-Ado. RESULTS: In MM cell lines both sensitive and resistant to conventional chemotherapies, 8-NH2-Ado is cytotoxic, with IC50 ranging from 300 nmol/L to 3 micromol/L. A mouse leukemic cell line lacking adenosine kinase activity was resistant to 8-NH2-Ado, indicating that phosphorylation of 8-NH2-Ado to its triphosphate form is required for cytotoxicity. A 4-hour incubation of MM cells with 10 micromol/L analogue resulted in an accumulation of >7 mmol/L 8-NH2-ATP with a parallel decline in the endogenous ATP levels. Accumulation of 8-NH2-ATP was dependent on both exogenous concentration of 8-NH2-Ado and incubation time. The accumulation of 8-NH2-ATP was accompanied by a decrease in both RNA and DNA synthesis. The mechanism of 8-NH2 Ado-mediated cytotoxicity was due to apoptosis as measured by an increase in Annexin V binding, a decrease in mitochondrial membrane potential, an increase in caspase activity, cleavage of caspase substrates, and an increase in cells with a sub-G1 DNA content. CONCLUSION: Based on these results, we conclude that 8-NH2 Ado may hold great potential as a therapeutic agent for the treatment of MM. PMID- 15542781 TI - Cyclooxygenase-2-dependent and -independent effects of celecoxib in pancreatic cancer cell lines. AB - Cyclooxygenase-2 (COX-2) is involved in inhibition of apoptosis, potentiation of cell growth, and angiogenesis and as such is a target for drug development. The COX-2 enzyme is frequently overexpressed in pancreatic cancer. The aim of this study was to determine the effects of celecoxib on the growth inhibition and induction of apoptosis by gemcitabine in pancreatic cancer cell lines. Baseline expression of COX-2 enzyme was determined by Western blot analysis in five human pancreatic cancer cell lines. Cells were treated with gemcitabine (100 nmol/L), celecoxib (1, 10, and 50 micromol/L), and the combination. No potentiation in growth inhibition was observed in MIAPaCa cells (low COX-2 expression). However, growth inhibition and apoptosis were significantly increased with celecoxib in the BxPC-3 cells that have a high COX-2 expression. Significant down-regulation of nuclear factor-kappaB activation was observed in BxPC-3 cells treated with celecoxib and gemcitabine. Moreover, down-regulation of COX-2 mRNA and protein expression was also observed in the BxPC-3 cells treated with the combination as compared with the untreated and the celecoxib-treated and gemcitabine-treated cell lines. We conclude that celecoxib potentiates gemcitabine-induced growth inhibition and apoptosis in pancreatic cell lines. In addition to inhibition of the COX-2 enzyme, the celecoxib and gemcitabine combination down-regulated nuclear factor-kappaB activation, which in turn may have contributed to the induction of apoptosis and the down-regulation of transcription of the COX-2 enzyme. PMID- 15542782 TI - Specific inhibition of cyclin-dependent kinase 4/6 by PD 0332991 and associated antitumor activity in human tumor xenografts. AB - PD 0332991 is a highly specific inhibitor of cyclin-dependent kinase 4 (Cdk4) (IC50, 0.011 micromol/L) and Cdk6 (IC50, 0.016 micromol/L), having no activity against a panel of 36 additional protein kinases. It is a potent antiproliferative agent against retinoblastoma (Rb)-positive tumor cells in vitro, inducing an exclusive G1 arrest, with a concomitant reduction of phospho Ser780/Ser795 on the Rb protein. Oral administration of PD 0332991 to mice bearing the Colo-205 human colon carcinoma produces marked tumor regression. Therapeutic doses of PD 0332991 cause elimination of phospho-Rb and the proliferative marker Ki-67 in tumor tissue and down-regulation of genes under the transcriptional control of E2F. The results indicate that inhibition of Cdk4/6 alone is sufficient to cause tumor regression and a net reduction in tumor burden in some tumors. PMID- 15542783 TI - Screening a combinatorial peptide library to develop a human glandular kallikrein 2-activated prodrug as targeted therapy for prostate cancer. AB - OBJECTIVE: Prostate cancer cells secrete the unique protease human glandular kallikrein 2 (hK2) that represents a target for proteolytic activation of cytotoxic prodrugs. The objective of this study was to identify hK2-selective peptide substrates that could be coupled to a cytotoxic analogue of thapsigargin, a potent inhibitor of the sarcoplasmic/endoplasmic reticulum calcium ATPase pump that induces cell proliferation-independent apoptosis through dysregulation of intracellular calcium levels. METHODS: To identify peptide sequence requirements for hK2, a combination of membrane-bound peptides (SPOT analysis) and combinatorial chemistry using fluorescence-quenched peptide substrates was used. Peptide substrates were then coupled to 8-O-(12[L-leucinoylamino]dodecanoyl)-8-O debutanoylthapsigargin (L12ADT), a potent analogue of thapsigargin, to produce a prodrug that was then characterized for hK2 hydrolysis, plasma stability, and in vitro cytotoxicity. RESULTS: Both techniques indicated that a peptide with two arginines NH2-terminal of the scissile bond produced the highest rates of hydrolysis. A lead peptide substrate with the sequence Gly-Lys-Ala-Phe-Arg-Arg (GKAFRR) was hydrolyzed by hK2 with a Km of 26.5 micromol/L, kcat of 1.09 s(-1), and a kcat/Km ratio of 41,132 s(-1) mol/L(-1). The GKAFRR-L12ADT prodrug was rapidly hydrolyzed by hK2 and was stable in plasma, whereas the GKAFRR-L peptide substrate was unstable in human plasma. The hK2-activated thapsigargin prodrug was not activated by cathepsin B, cathepsin D, and urokinase but was an excellent substrate for plasmin. The GKAFRR-L12ADT was selectively cytotoxic in vitro to cancer cells in the presence of enzymatically active hK2. CONCLUSION: The hK2 activated thapsigargin prodrug represents potential novel targeted therapy for prostate cancer. PMID- 15542784 TI - Secretory phospholipase A2 as a tumor-specific trigger for targeted delivery of a novel class of liposomal prodrug anticancer etherlipids. AB - The use of many common clinically relevant chemotherapeutics is often limited due to insufficient delivery to the tumor and dose-limiting systemic toxicities. Therefore, therapeutics that specifically target tumor cells and are nontoxic to normal cells are required. Here, we report the development of a novel class of liposomes composed of lipid prodrugs, which use the increased secretory phospholipase A2 type IIA (sPLA2) activity of the tumor microenvironment as a trigger for the release of anticancer etherlipids (AEL). Treatment of sPLA2 secreting tumor cells in vitro with liposomes consisting of proAELs resulted in growth inhibition comparable with addition of the AELs alone. Using a specific sPLA2 inhibitor, we showed the low cytotoxicity of the nonhydrolyzed proAEL liposomes and have proven the sPLA2 dependency of the activation of proAELs to cytotoxic AELs. In addition, we showed that our proAEL liposomes circumvent the inherent hemolytic toxicities associated with the use of etherlipids, thereby allowing i.v. administration of such therapeutics as nontoxic prodrug liposomes. Furthermore, using a sPLA2-secreting human colon cancer xenograft model, we showed that the proAEL liposomes are capable of inducing a tumor growth delay in vivo. Taken together, these data support the validity of this novel tumor selective liposomal prodrug delivery strategy. This new approach also provides a promising system for tumor-selective delivery and release of conventional chemotherapeutics encapsulated in the sPLA2-degradable prodrug liposomes. PMID- 15542785 TI - Modulation of p53 in 7,12-dimethylbenz[a]anthracene-induced skin tumors by diallyl sulfide in Swiss albino mice. AB - Allium vegetables have been shown to have beneficial health effects against several chronic diseases including cancer. Diallyl sulfide (DAS), an organosulfur compound present in garlic, is well known for its chemopreventive properties in several tumor models. The pharmacologic role of DAS in prevention and treatment of cancer is well documented in the literature, but its molecular mechanism of action is not yet well defined. In the present study, modulation in p53 expression by topical application of DAS was recorded in 7,12 dimethylbenz[a]anthracene (DMBA)-induced skin tumors in Swiss albino mice. Western blot analysis and immunohistochemical protein detection, combined with multivariable flow cytometry, show that DAS application induces the expression of the wild-type (wt) p53 and down-regulates the expression of mutant (mut) p53. Immunoblotting analysis of tumors showed significant increase in levels of wtp53 by DAS application, whereas for mutp53 the DMBA-induced levels of protein were found to reduce to near normal levels with DAS application. The quantitative analysis of immunostained skin/tumor sections using image analysis and quantitative stereology showed 66.6% and 54.2% increases in wtp53 levels and 53.4% and 44.3% decreases in mutp53 levels in animals where DAS was applied 1 hour prior to or 1 hour after DMBA application, respectively. Flow cytometric analysis further confirmed modulation of wtp53 and mutp53 protein in DAS supplemented tumors. The increase in the expression of wt tumor suppressor gene protein p53 was accompanied by elevation of the levels of cyclin-dependent kinase inhibitor p21/waf1. The percentage increase in the levels of p21/waf1 was found to be 72.9% and 61.3%, respectively, in DAS-supplemented groups before and after administration. These results thus show that DAS is a potential chemopreventive agent capable of modulating and regulating the tumor suppressor p53 along with its downstream effective molecule, p21/waf1. Thus, DAS can be a potential chemopreventive agent against skin tumor development. PMID- 15542786 TI - Disruption of WT1 gene expression and exon 5 splicing following cytotoxic drug treatment: antisense down-regulation of exon 5 alters target gene expression and inhibits cell survival. AB - Deregulated expression of the Wilms' tumor gene (WT1) has been implicated in the maintenance of a malignant phenotype in leukemias and a wide range of solid tumors through interference with normal signaling in differentiation and apoptotic pathways. Expression of high levels of WT1 is associated with poor prognosis in leukemias and breast cancer. Using real-time (Taqman) reverse transcription-PCR and RNase protection assay, we have shown up-regulation of WT1 expression following cytotoxic treatment of cells exhibiting drug resistance, a phenomenon not seen in sensitive cells. WT1 is subject to alternative splicing involving exon 5 and three amino acids (KTS) at the end of exon 9, producing four major isoforms. Exon 5 splicing was disrupted in all cell lines studied following a cytotoxic insult probably due to increased exon 5 skipping. Disruption of exon 5 splicing may be a proapoptotic signal because specific targeting of WT1 exon 5 containing transcripts using a nuclease-resistant antisense oligonucleotide (ASO) killed HL60 leukemia cells, which were resistant to an ASO targeting all four alternatively spliced transcripts simultaneously. K562 cells were sensitive to both target-specific ASOs. Gene expression profiling following treatment with WT1 exon 5-targeted antisense showed up-regulation of the known WT1 target gene, thrombospondin 1, in HL60 cells, which correlated with cell death. In addition, novel potential WT1 target genes were identified in each cell line. These studies highlight a new layer of complexity in the regulation and function of the WT1 gene product and suggest that antisense directed to WT1 exon 5 might have therapeutic potential. PMID- 15542787 TI - The role of nuclear Y-box binding protein 1 as a global marker in drug resistance. AB - Gene expression can be regulated by nuclear factors at the transcriptional level. Many such factors regulate MDR1 gene expression, but what are the sequence elements and transcription factors that control the basal and inducible expression of this gene? The general principles through which transcription factors participate in drug resistance are now beginning to be understood. Here, we review the factors involved in the transcriptional regulation of the MDR1 gene. In particular, we focus on the transcription factor Y-box binding protein 1 and discuss the possible links between Y-box binding protein 1 expression and drug resistance in cancer, which are mediated by the transmembrane P-glycoprotein or non-P-glycoprotein. PMID- 15542788 TI - Biological impediments to monoclonal antibody-based cancer immunotherapy. AB - The ability of antibodies to exploit antigenic differences between normal and malignant tissues and to exact a variety of antitumor responses offers significant advantages to conventional forms of therapy. Several monoclonal antibodies (mAb) have already proved to be relatively well tolerated and effective for the treatment of many different malignant diseases. However, mAbs must overcome substantial obstacles to reach antigens presented on target cells to be of therapeutic value. Intravenously administered antibodies must avoid host immune response and contend with low or heterogeneous expression of antigen on tumor cells. Antibodies must also overcome significant physical barriers en route to a solid tumor mass, including the vascular endothelium, stromal barriers, high interstitial pressure, and epithelial barriers. Here we review the application and evolution of mAbs as effective forms of treatment, with particular attention to the barriers and impediments to successful treatment and discuss strategies to overcome these barriers and improve the efficacy of mAb-based therapy. PMID- 15542789 TI - Cytochrome P450 enzymes and tumor therapy. PMID- 15542790 TI - Oleic acid inhibits alveolar fluid reabsorption: a role in acute respiratory distress syndrome? AB - Levels of oleic acid (OA) are elevated in plasma and bronchoalveolar lavage fluids of patients with acute respiratory distress syndrome (ARDS). OA is also widely used to provoke edema, by unknown mechanisms, in experimental models of ARDS. We investigated the impact of intravascularly applied OA on epithelial lining fluid balance. OA (25 microM) dramatically blocked active transepithelial (22)Na(+) transport (by 92%) in an isolated, ventilated, and perfused rabbit lung model, provoking alveolar edema, assessed by increases in lung weight and epithelial lining fluid volume. OA did not alter epithelial permeability, measured by [(3)H]mannitol and fluorescently labeled albumin flux, but did increase endothelial permeability, assessed by capillary filtration coefficient. In A549 cells, OA completely blocked amiloride-sensitive sodium currents measured by patch clamp, and also largely abrogated ouabain-sensitive Na(+),K(+)-ATPase mediated (86)Rb(+) uptake. Although OA did not alter epithelial sodium channel or Na(+),K(+)-ATPase surface expression, it covalently associated with both molecules and directly, dramatically, and dose-dependently inhibited the catalytic activity of purified Na(+),K(+)-ATPase. Therefore, OA impaired the two essential transepithelial active sodium transport mechanisms of the lung, and could thus promote alveolar edema formation and prevent edema resolution, thereby contributing to the development of ARDS. PMID- 15542791 TI - Association of habitual snoring with glucose and insulin metabolism in nonobese Korean adult men. AB - Habitual snoring is associated with cardiovascular morbidity and mortality, and metabolic abnormalities such as impaired glucose homeostasis. Many studies were performed in obese Western populations. The purpose of this study was to examine the association of habitual snoring with glucose and insulin metabolism in nonobese Korean men who were free of diabetes and hypertension. A total of 2,719 men ages 40-69 years from the Korean Health and Genome Study participated in this study. Information of snoring frequency was obtained by a questionnaire and glucose and insulin levels during oral glucose tolerance test were measured. Repeated measures analysis of variance detected significant differences in the changing patterns of glucose and insulin levels at 1 hour and 2 hours between habitual snorers and nonhabitual snorers, whereas there were no significant differences in fasting blood glucose and insulin levels. Multivariate logistic regression analyses revealed that habitual snoring was independently associated with elevated 1-hour and 2-hour glucose levels and a 2-hour insulin level, respectively. The present data suggest that habitual snoring may affect glucose insulin metabolism, independent of diabetes and hypertension, even in nonobese Korean middle-age men. Further prospective studies are needed to examine the causal relationship between habitual snoring and insulin resistance or glucose intolerance. PMID- 15542792 TI - Montelukast reduces asthma exacerbations in 2- to 5-year-old children with intermittent asthma. AB - The PREVIA study was designed to investigate the role of montelukast, a leukotriene receptor antagonist, in the prevention of viral-induced asthma exacerbations in children aged 2 to 5 years with a history of intermittent asthma symptoms. The study was a 12-month multicenter, double-blind, parallel-group study of patients with asthma exacerbations associated with respiratory infections and minimal symptoms between episodes. Patients were randomized to receive oral montelukast 4 or 5 mg (depending on age) (n = 278) or placebo (n = 271) once per day for 12 months. Caregivers recorded children's symptoms, beta agonist use, and health care resource use in a diary card. Over 12 months of therapy, montelukast significantly reduced the rate of asthma exacerbations by 31.9% compared with placebo. The average rate of exacerbation episodes per patient was 1.60 episodes per year on montelukast compared with 2.34 episodes on placebo. Montelukast also delayed the median time to first exacerbation by approximately 2 months (p = 0.024), and the rate of inhaled corticosteroid courses (p = 0.027) compared with placebo. Montelukast effectively reduced asthma exacerbations in 2- to 5-year-old patients with intermittent asthma over 12 months of treatment and was generally well tolerated. PMID- 15542793 TI - Two-year cognitive, emotional, and quality-of-life outcomes in acute respiratory distress syndrome. AB - Acute respiratory distress syndrome (ARDS) has a high mortality and is associated with significant morbidity. Prior outcome studies have focused predominant on short-term outcomes (6-12 months). We assessed longitudinal neurocognitive, emotional, and quality of life in ARDS survivors at hospital discharge, and 1 and 2 years after hospital discharge using neuropsychologic tests and emotional and quality-of-life questionnaires. Neurocognitive sequelae occurred in 73% (54 of 74) of ARDS survivors at hospital discharge, 46% (30 of 66) at 1 year, and 47% (29 of 62) at 2 years. ARDS survivors report moderate to severe depression (16% and 23%) and anxiety (24% and 23%) at 1 and 2 years, respectively. The ARDS survivors had decreased quality of life, with the physical domains improving at 1 year, with no additional change at 2 years. Role emotional, pain, and general health did not change from hospital discharge to 2 years. Mental health improved during the first year and declined at 2 years. ARDS results in significant neurocognitive and emotional morbidity and decreased quality of life that persists at least 2 years after hospital discharge. ARDS can cause significant long-term, brain-related morbidity manifest by neurocognitive impairments and decreased quality of life. PMID- 15542794 TI - Hyperventilation with cold versus dry air in 2- to 5-year-old children with asthma. AB - Cold air challenge (CACh) has been shown to discriminate between children with asthma and healthy young children. Hyperventilation with dry room-temperature air is a simplified alternative. We compared responsiveness in young children with asthma between two standardized, single-step protocols: dry air challenge (DACh) performed as 6 minutes of eucapnic hyperventilation with dry room-temperature air and CACh as 4 minutes of hyperventilation. Response was measured as specific airway resistance by whole-body plethysmography and expressed as change from baseline in numbers of within-subject SDs (SDw). The challenge sequence was randomly assigned. A comparator challenge was performed 1 hour later if the first challenge gave a change of 3 SDw or more. Forty 2- to 5-year-old children with asthma were included. Responsiveness to cold versus dry air showed significant, but weak, correlation (r(2) = 0.34, p < 0.0001), but responsiveness to CACh exceeded DACh (7.6 vs. 5.4 SDw, p < 0.02). CACh seemed to induce reduction in response to the following DACh (p < 0.01), whereas no such reduction was seen after DACh. CONCLUSION: Responsiveness to CACh exceeded responsiveness to DACh, and CACh seemed to induce refractoriness in contrast to DACh, probably because of the additional stimulus from airway cooling. This finding suggests CACh as the preferred method of challenge. PMID- 15542795 TI - Prospective surveillance for Pseudomonas aeruginosa cross-infection at a cystic fibrosis center. AB - We have performed a 4-year prospective surveillance for Pseudomonas aeruginosa cross-infection at a large regional adult cystic fibrosis center. Despite purpose built facilities in a new building and the practice of strict hygiene, P. aeruginosa cross-infection has continued. In contrast, individuals segregated from the cohort of patients with chronic P. aeruginosa infection but who attend the same center have not acquired infection with transmissible P. aeruginosa strains. Simple infection control measures alone do not prevent the spread of transmissible P. aeruginosa strains between individuals with cystic fibrosis. However, in our clinic patient segregation effectively controlled spread of such strains. PMID- 15542796 TI - Clinical oncology 2004: the president's report. PMID- 15542797 TI - Increasing access to clinical cancer trials and emerging technologies for minority populations: the Native American Project. PMID- 15542798 TI - Immunologic and clinical outcomes of vaccination with a multiepitope melanoma peptide vaccine plus low-dose interleukin-2 administered either concurrently or on a delayed schedule. AB - PURPOSE: A phase II trial was performed to test whether systemic low-dose interleukin-2 (IL-2) augments T-cell immune responses to a multipeptide melanoma vaccine. Forty patients with resected stage IIB-IV melanoma were randomly assigned to vaccination with four gp100- and tyrosinase-derived peptides restricted by human leukocyte antigen (HLA) -A1, HLA-A2, and HLA-A3, and a tetanus helper peptide plus IL-2 administered daily either beginning day 7 (group 1), or beginning day 28 (group 2). PATIENTS AND METHODS: T-cell responses were assessed by an interferon gamma ELIspot assay in peripheral blood lymphocytes (PBL) and in a lymph node draining a vaccination site (sentinel immunized node [SIN]). Patients were followed for disease-free and overall survival. RESULTS: T cell responses to the melanoma peptides were observed in 37% of PBL and 38% of SINs in group 1, and in 53% of PBL and 83% of SINs in group 2. The magnitude of T cell response was higher in group 2. The tyrosinase peptides DAEKSDICTDEY and YMDGTMSQV were more immunogenic than the gp100 peptides YLEPGPVTA and ALLAVGATK. T-cell responses were detected in the SINs more frequently, and with higher magnitude, than responses in the PBL. Disease-free survival estimates at 2 years were 39% (95% CI, 18% to 61%) for group 1, and 50% (95% CI, 28% to 72%) for group 2 (P = .32). CONCLUSION: The results of this study support the safety and immunogenicity of a vaccine composed of four peptides derived from gp100 and tyrosinase. The low-dose IL-2 regimen used for group 1 paradoxically diminishes the magnitude and frequency of cytotoxic T lymphocyte responses to these peptides. PMID- 15542799 TI - Adjuvant chemotherapy for resected adenocarcinoma of the esophagus, gastro esophageal junction, and cardia: phase II trial (E8296) of the Eastern Cooperative Oncology Group. AB - PURPOSE: To evaluate the effect of postoperative paclitaxel and cisplatin on 2 year survival in patients with completely resected adenocarcinoma of the distal esophagus, gastro-esophageal (GE) junction, and cardia. PATIENTS AND METHODS: We conducted a multicenter phase II trial. Patients had pathologically staged T2 node-positive to T3-4, any node status adenocarcinoma of the distal esophagus, GE junction, or gastric cardia with negative margins (R0). Treatment consisted of four cycles of paclitaxel 175 mg/m2 intravenously (i.v.) over 3 hours followed by cisplatin 75 mg/m2 i.v. every 21 days. A positive outcome was considered to be an improvement in 2-year survival rate by > or = 20% compared to historic controls. RESULTS: Fifty-nine patients were recruited from 20 centers. Of 55 eligible patients, 49 (89%) had lymph node involvement. Forty-six patients (84%) completed all four cycles. Of the total 59 patients, 31 (56%) developed grade 3 or 4 toxicity with leukopenia/neutropenia, nausea/vomiting, and metabolic toxicities were most common. The median follow-up for surviving patients was 4 years. At 2 years, 33 patients were alive and 22 were dead, with a survival rate of 60% (95% CI, 46% to 73%; one-sided P = .0008 compared with the historic controls). CONCLUSION: Our data suggest that adjuvant paclitaxel and cisplatin may improve survival in R0 resected patients with locally advanced adenocarcinoma of the distal esophagus, GE junction, and cardia. These results warrant further testing in randomized trials. PMID- 15542800 TI - Wilms' tumor in adults: results of the Society of Pediatric Oncology (SIOP) 93 01/Society for Pediatric Oncology and Hematology (GPOH) Study. AB - PURPOSE: In the Society of Pediatric Oncology (SIOP) 93-01 study, 30 patients older than 16 years were found to have Wilms' tumor. They were treated according to the pediatric protocol and were analyzed for clinical presentation, stage distribution, and prognosis. PATIENTS AND METHODS: Patient age ranged from 16 to 62 years (median, 25.4 years). Tumor stages were defined according to SIOP, and treatment was risk-adapted according to SIOP 93-01/Society for Pediatric Oncology and Hematology (GPOH) protocol. The patients were evaluated with regard to response, toxicity, and prognosis. Specimens of all tumors were centrally reviewed. RESULTS: Ten patients (33%) had metastatic disease at the time of diagnosis (liver, four patients; lung, three patients; liver and lung, three patients). The local stage distribution showed a predominance of higher stages (stage I, eight patients; stage IIN-, three patients; stage IIN+, four patients; stage III, 15 patients). Histologic studies revealed intermediate-risk in 23 of 30 tumors; two tumors were classified as high-risk; and three tumors were clear cell sarcomas. Two of 30 patients showed a nephroblastoma and a renal cell carcinoma simultaneously in the same kidney. A complete remission was achieved in 24 patients; four patients relapsed after complete remission; and three of them reached a second remission with further treatment. Event-free survival was 57%, with an overall survival of 83% (median observation time, 4 years). CONCLUSION: Adults can be cured in a high percentage by a multimodal treatment according to pediatric protocols. Toxicity is higher than in children, but acceptable in view of the high remission rate. PMID- 15542801 TI - Insulin, physical activity, and caloric intake in postmenopausal women: breast cancer implications. AB - PURPOSE: Increased physical activity and programs to reduce body mass index (BMI) with both increased physical activity and decreased caloric intake have been proposed to reduce insulin as a potential mediator of breast cancer and other chronic diseases. However, there are few data on the relative contribution of physical activity, caloric intake, and BMI to fasting insulin levels. MATERIALS AND METHODS: An ethnically diverse subsample of 2,996 mostly healthy postmenopausal women with no prior cancer history was randomly identified from the 161,809 participants in the Women's Health Initiative clinical trials and observational study. Information was collected on diet, recreational physical activity, and anthropometrics including BMI. Fasting insulin levels were determined. Using a cross-sectional design, insulin levels were then compared across quintiles of caloric intake and physical activity in linear regression model analyses controlled for BMI and other factors. RESULTS: Lower BMI (P < .0001), higher levels of physical activity (P < .0001), and lower caloric intake (P < .02) were all independently associated with significantly lower mean fasting insulin levels throughout the range of observed values. Insulin levels of 8.74 microU/mL +/- 4.16 SD were seen in the highest physical activity and lowest caloric intake quintile compared with insulin levels of 15.08 microU/mL +/- 16.32 SD in the lowest physical activity and highest caloric intake quintile (P < .0001). CONCLUSION: These findings suggest that reduction in BMI achieved by increasing physical activity, reducing caloric intake, or both, should lower insulin levels, providing support for clinical trials evaluating insulin level change and breast cancer risk. PMID- 15542802 TI - Prevalence of KIT expression in human tumors. AB - PURPOSE: KIT is a target for imatinib mesylate (Gleevec; Novartis Pharma, Basel, Switzerland). Gastrointestinal stromal tumors (GISTs) express KIT and respond favorably to imatinib therapy. To determine other tumors in which such a molecular targeted therapy might be indicated, we investigated KIT expression in different human tumor types. Because recent studies in GISTs suggest that KIT activating mutations predict response to imatinib therapy, we also sequenced a subset of positive tumors. MATERIALS AND METHODS: More than 3,000 tumors from more than 120 different tumor categories were analyzed by immunohistochemistry in a tissue microarray format. Seven commercially available anti-KIT antibodies were initially evaluated. The antibody A4502 (DAKO) was selected for analysis because of a high frequency of positivity in GIST and low staining background in other tissues. To determine the frequency of KIT mutations in various tumor types, the exons 2, 8, 9, 11, 13, and 17 (where mutations previously were reported) were sequenced in 36 tumors with strong KIT expression. RESULTS: KIT positivity was detected in 28 of 28 GISTs (100%), 42 of 50 seminomas (84%), 34 of 52 adenoid cystic carcinomas (65%), 14 of 39 malignant melanomas (35%), and eight of 47 large-cell carcinomas of the lung (17%), as well as in 47 additional tumor types. KIT mutations were found in six of 12 analyzed GISTs, but only in one of 24 other tumors. CONCLUSION: The results suggest that KIT expression occurs infrequently in most tumor types and that, with the exception of GISTs, KIT gene mutations are rare in immunohistochemically KIT-positive tumors. PMID- 15542803 TI - Phase II study of CT-2103 in patients with recurrent epithelial ovarian, fallopian tube, or primary peritoneal carcinoma. AB - PURPOSE: To evaluate the safety and efficacy of CT-2103, a novel conjugate of paclitaxel and poly-L-glutamic acid, in heavily pretreated patients with recurrent ovarian, fallopian tube, or peritoneal cancer. PATIENTS AND METHODS: Ninety-nine patients with measurable disease received intravenous CT-2103 at 175 mg/m2 of conjugated paclitaxel over 10 minutes every 3 weeks without routine premedications. Platinum-sensitive (n = 42) and platinum-refractory or platinum resistant patients (n = 57) were enrolled. Thirty-nine patients (39%) had received one or two prior regimens, and 60 patients (61%) had received between three and 12 regimens. RESULTS: In 99 patients, the median number of cycles was three (range, one to 14 cycles). The response rate (RR) for all patients was 10% (10 of 99 patients), with median time to disease progression (TTP) of 2 months. The RR (partial response) in platinum-sensitive and platinum-resistant patients was 14% (six of 42 patients) and 7% (four of 57 patients), respectively. In patients with only one or two prior regimens, the RR in platinum-sensitive and platinum-resistant patients was 28% (five of 18 patients) and 10% (two of 21 patients), with a median TTP of 4 and 2 months, respectively. Grade 2 (15 patients) or 3 (15 patients) neuropathy was reported in 30 patients (30%). Grade 2 hypersensitivity occurred in eight patients (8%) who were subsequently treated with premedications; one patient had grade 3 hypersensitivity and was removed. Grade 2 alopecia was absent. CONCLUSION: CT-2103 is active in patients with recurrent ovarian cancer. Neurotoxicity in these heavily pretreated patients was more frequent than predicted from phase I trials. Further study to define toxicity and efficacy in patients with less prior therapy is ongoing. PMID- 15542804 TI - High-dose therapy and autologous hematopoietic stem-cell transplantation for recurrent or refractory pediatric Hodgkin's disease: results and prognostic indices. AB - PURPOSE: To evaluate the outcome of pediatric patients with refractory or relapsed Hodgkin's disease (HD) who undergo high-dose therapy and autologous hematopoietic stem-cell transplantation (AHSCT). PATIENTS AND METHODS: From 1989 to 2001, 41 pediatric patients with relapsed or primary refractory HD underwent high-dose therapy followed by AHSCT according to one of four autologous transplantation protocols at Stanford University Medical Center (Stanford, CA). Pretreatment factors were analyzed by univariate and multivariate analysis for prognostic significance for 5-year overall survival (OS), event-free survival (EFS), and progression-free survival (PFS). RESULTS: At a median follow-up of 4.2 years (range, 0.7 to 11.9 years), the 5-year OS, EFS, and PFS rates were 68%, 53%, and 63%, respectively. Multivariate analysis determined the following three factors to be significant predictors of poor OS and EFS: extranodal disease at first relapse, presence of mediastinal mass at time of AHSCT, and primary induction failure. Two of these factors also predicted for poor PFS (extranodal disease at time of first relapse and presence of mediastinal mass at time of transplantation). CONCLUSION: More than half of children with relapsed or refractory HD can be successfully treated with the combination of high-dose therapy and AHSCT, confirming the efficacy of this approach. Further investigation is now required to determine the optimal timing of AHSCT, as well as to develop alternative regimens for those patients with factors prognostic for poor outcome after AHSCT. PMID- 15542805 TI - Risk-adapted, combined-modality therapy with VAMP/COP and response-based, involved-field radiation for unfavorable pediatric Hodgkin's disease. AB - PURPOSE: To evaluate the efficacy of vinblastine, doxorubicin, methotrexate, and prednisone (VAMP) and cyclophosphamide, vincristine, and procarbazine (COP) chemotherapy and response-based, involved-field radiation, a combined-modality regimen that limits doses of alkylating agents, anthracyclines, and radiation, in children with advanced and unfavorable Hodgkin's disease. PATIENTS AND METHODS: From 1993 to 2000, 159 children and adolescents with unfavorable Hodgkin's disease received three alternating cycles (total of six cycles) of VAMP/COP chemotherapy followed by response-based, involved-field radiation therapy: 15 Gy was administered to patients achieving a complete response, and 25.5 Gy was administered to those achieving a partial response after the first two cycles of chemotherapy and to all sites of bulky lymphadenopathy. Unfavorable disease was defined as clinical stage I and II with bulky peripheral nodal disease greater than 6 cm, initial bulky mediastinal mass 33% or more of the intrathoracic diameter, and/or "B" symptoms and all stage III and IV. RESULTS: Study enrollment was closed after an interim analysis estimated a 5-year event-free survival (EFS) rate below a predefined level. Disease presentation was localized (stage I/II) in 77 patients (48.4%) and advanced (stage III/IV) in 82 patients (51.6%). At a median follow-up of 5.8 years (range, 1.3 to 10.0 years), 38 patients had events, including relapse/progression (n = 35), second malignancy (n = 2), and accidental death (n = 1); nine relapses (25.7%) occurred greater than 4 years from diagnosis. Five-year survival and EFS estimates are 92.7% +/- 2.5% and 75.6% +/- 4.1%, respectively. CONCLUSION: Risk-adapted combined-modality therapy with VAMP/COP and response-based, involved-field radiation therapy results in an unsatisfactory outcome for pediatric patients with unfavorable presentations of Hodgkin's disease. PMID- 15542806 TI - White matter lesions detected by magnetic resonance imaging after radiotherapy and high-dose chemotherapy in children with medulloblastoma or primitive neuroectodermal tumor. AB - PURPOSE: White matter lesions (WMLs) have been described as a delayed effect of cranial irradiation in children with brain tumors, or a transient subacute effect characterized by an intralesional or perilesional reaction. We report the occurrence of subacute WMLs detected by magnetic resonance imaging (MRI) in children treated for medulloblastoma or primitive neuroectodermal tumor (PNET) and document the associated clinical, radiologic, and neurocognitive findings. PATIENTS AND METHODS: Among 134 patients with medulloblastoma or supratentorial PNET treated prospectively with risk-adjusted craniospinal irradiation and conformal boost to the tumor bed, followed by four high-dose chemotherapy (HDC) cycles with stem-cell rescue, 22 developed WMLs on T1-weighted imaging with and without contrast and/or T2-weighted imaging on MRI. Patients had > or = 12 months of follow-up. Neurocognitive assessments included intelligence quotient (IQ) tests and tests of academic achievement. RESULTS: Twenty-two patients developed WMLs at a median of 7.8 months after starting therapy (range, 1.9 to 13.0 months). Lesions were predominantly in the pons (n = 8) and cerebellum (n = 6). Sixteen patients (73%) had WML resolution at a median of 6.2 months (range, 1.68 to 23.5 months) after onset; two patients developed necrosis and atrophy. Three developed persistent neurologic deficits. Cumulative incidence of WMLs at 1 year was 15% +/- 3%. Patients with WMLs had a significant decline in estimated IQ ( 2.5 per year; P = .03) and math (-4.5 per year; P = .003) scores. CONCLUSION: WMLs in medulloblastoma or PNET patients treated with conformal radiotherapy and HDC are typically transient and asymptomatic, and may mimic early tumor recurrence. A minority of patients with WMLs develop permanent neurologic deficits and imaging changes. Overall, the presence of WMLs is associated with greater neurocognitive decline. PMID- 15542807 TI - Effects of pretransplantation treatment with rituximab on outcomes of autologous stem-cell transplantation for non-Hodgkin's lymphoma. AB - PURPOSE: To analyze the effects of preautografting treatment with rituximab (R) on stem-cell mobilization, post-transplantation complications, engraftment, disease-free survival, and overall survival in patients with non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Single-institution retrospective comparative outcome analysis in a cohort of 273 relapsed chemosensitive NHL patients of whom 127 (47%) received R pretransplantation. RESULTS: R was administered a median of 3 months before autologous transplantation. When compared to the nonrituximab group, R patients were older (56 v 50 years; P < .001), and had delays in post-transplantation platelets recovery (39 v 27 days; P = .001). Pretransplantation R did not affect stem-cell mobilization, post transplantation early complications, duration of hospitalization, or mortality rates at days 30 and 100. In contrast to patients with low-grade NHL, both disease-free and overall survival rates were significantly better when R was included in the pretransplantation salvage therapy for patients with intermediate grade NHL. CONCLUSION: In this large, single-center retrospective analysis, pretransplantation treatment with R was associated with improved survival in patients with intermediate-grade NHL, at the price, however, of a delay in platelet engraftment. PMID- 15542808 TI - Cohort analysis of patients with localized, high-risk, extremity soft tissue sarcoma treated at two cancer centers: chemotherapy-associated outcomes. AB - PURPOSE: Patients with American Joint Committee on Cancer stage III soft tissue sarcoma (STS) have high risks of distant recurrence and death. The role of chemotherapy for these patients remains controversial despite several randomized trials and a meta-analysis. METHODS: We reviewed the treatments and outcomes of 674 consecutive adult patients presenting with primary stage III extremity STS between 1984 and 1999. Pre- or postoperative doxorubicin-based chemotherapy was used in a nonrandomized fashion in approximately half of this high-risk population. The objective of this review was to evaluate the impact of chemotherapy while accounting for known prognostic variables. RESULTS: Among 674 patients, 338 (50%) were treated with local therapy only, and 336 (50%) were treated with local therapy plus chemotherapy. The median follow-up for survivors was 6.1 years. Five-year local and distant recurrence-free interval probabilities were 83% and 56%, respectively, for the two groups combined. The 5-year disease specific survival (DSS) rate was 61%. Cox regression analyses showed a time varying effect associated with chemotherapy. During the first year, the hazard ratio associated with DSS for patients treated with chemotherapy versus no chemotherapy was 0.37 (95% CI, 0.20 to 0.69; P = .002). Thereafter, this hazard ratio was 1.36 (95% CI, 1.02 to 1.81; P = .04). CONCLUSION: It seems that the clinical benefits associated with doxorubicin-based chemotherapy in patients with high-risk extremity STS are not sustained beyond 1 year. These results suggest that caution should be used in the interpretation of randomized clinical trials of adjuvant chemotherapy that seem to demonstrate clinical benefits with relatively short-term follow-up. PMID- 15542809 TI - Prognostic factors in resected stage I non-small-cell lung cancer: a multivariate analysis of six molecular markers. AB - PURPOSE: To analyze the prognostic significance of six molecular biomarkers (death-associated protein kinase [DAPK] promoter methylation, interleukin-10 [IL 10] protein expression, cyclooxygenase-2 [COX-2] mRNA expression, human telomerase reverse transcriptase catalytic subunit [hTERT] mRNA expression, retinoic acid receptor-beta [RAR-beta] mRNA expression, and K-ras mutational status) in stage I non-small-cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: Biomarker analyses were performed on tumors from 94 patients with stage I NSCLC who underwent surgical resection at our institution. A minimum follow-up period of 5 years was required. DAPK methylation was assessed by methylation specific polymerase chain reaction (PCR). RAR-beta, COX-2, and hTERT mRNA levels were determined by in situ hybridization with digoxigenin-labeled antisense riboprobes. K-ras mutation status was determined by the PCR-primer introduced restriction with enrichment for mutant alleles method. IL-10 protein expression was analyzed by immunohistochemistry using a polyclonal antihuman IL-10 antibody. Cancer-specific survival was analyzed with a Cox proportional hazards model. To identify independent prognostic factors, a stepwise selection method was used. RESULTS: DAPK methylation, IL-10 lack of expression, COX-2 expression, hTERT expression, RAR-beta expression, and K-ras mutations were observed in 46.8%, 29.8%, 59.6%, 34.0%, 23.4%, and 34.0% of patients, respectively. In the final model, DAPK methylation and IL-10 lack of expression were significant negative prognostic factors for cancer-specific survival, whereas COX-2 expression was of borderline significance. CONCLUSION: In this cohort of resected stage I NSCLC patients, molecular markers that independently predict cancer-specific survival have been identified. The prognostic roles of DAPK methylation, IL-10, and other biomarkers in NSCLC merit further investigation. PMID- 15542810 TI - Colorectal cancer with mutation in BRAF, KRAS, and wild-type with respect to both oncogenes showing different patterns of DNA methylation. AB - PURPOSE: BRAF mutations are common in sporadic colorectal cancers (CRCs) with a DNA mismatch repair (MMR) deficiency that results from promoter methylation of hMLH1, whereas KRAS mutations are common in MMR proficient CRCs associated with promoter methylation of MGMT. The aim of this study was to further investigate the link between genetic alterations in the RAS/RAF/ERK pathway and an underlying epigenetic disorder. PATIENTS AND METHODS: Activating mutations of BRAF and KRAS were identified and correlated with promoter methylation of 11 loci, including MINT1, MINT2, MINT31, CACNA1G, p16(INK4a), p14(ARF), COX2, DAPK, MGMT, and the two regions in hMLH1 in 468 CRCs and matched normal mucosa. RESULTS: BRAF V599E mutations were identified in 21 (9%) of 234 CRCs, and KRAS mutations were identified in 72 (31%) of 234 CRCs. Mutations in BRAF and KRAS were never found in the same tumor. CRCs with BRAF mutations showed high-level promoter methylation in multiple loci, with a mean number of methylated loci of 7.2 (95% CI, 6.6 to 7.9) among 11 loci examined (P < .0001). Tumors with KRAS mutations showed low-level promoter methylation, and CRCs with neither mutation showed a weak association with promoter methylation, with an average number of methylated loci of 1.8 (95% CI, 1.5 to 2.1) and 1.0 (95% CI, 0.79 to 1.3), respectively. CONCLUSION: In CRC, the methylation status of multiple promoters can be predicted through knowledge of BRAF and, to a lesser extent, KRAS activating mutations, indicating that these mutations are closely associated with different patterns of DNA hypermethylation. These changes may be important events in colorectal tumorigenesis. PMID- 15542811 TI - The additional value of chemotherapy to radiotherapy in locally advanced nasopharyngeal carcinoma: a meta-analysis of the published literature. AB - PURPOSE: The purpose of this meta-analysis was to determine the additional value of neoadjuvant, concurrent, and/or adjuvant chemotherapy to radiation in the treatment of locally advanced nasopharyngeal carcinoma (NPC) with regard to the overall survival (OS) and the incidence of local-regional recurrences (LRR) and distant metastases (DM). PATIENTS AND METHODS: To be eligible, full published studies had to deal with biopsy-proven NPC and have patients randomly assigned to receive conventional radiotherapy (66 to 70 Gy in 7 weeks) or radiotherapy combined with chemotherapy. RESULTS: Ten randomized clinical studies were identified, including 2,450 patients. The pooled hazard ratio (HR) of death for all studies was 0.82 (95% CI, 0.71 to 0.95; P = .01) corresponding to an absolute survival benefit of 4% after 5 years. Three categories of trials were defined according to the sequence of chemotherapy, including neoadjuvant chemotherapy, at least concomitant chemoradiotherapy, and adjuvant chemotherapy. A significant interaction term (P = .02) was found among these three categories. The largest effect was found for concomitant chemotherapy, with a pooled HR of 0.48 (95% CI, 0.32 to 0.72), which corresponds to a survival benefit of 20% after 5 years. Comparable results were found for the incidence of LRR and DM. CONCLUSION: The results of this study indicate that concomitant chemotherapy in addition to radiation is probably the most effective way to improve OS in NPC. PMID- 15542812 TI - Enrollment of elderly patients in clinical trials for cancer drug registration: a 7-year experience by the US Food and Drug Administration. AB - PURPOSE: To analyze the age-related enrollment of cancer patients onto registration trials of new drugs or new indications approved by the US Food and Drug Administration from 1995 to 2002. PATIENTS AND METHODS: This study involved retrospective analyses of demographic data of cancer patients enrolled onto registration trials. The data on 28,766 cancer patients from 55 registration trials were analyzed according to age distributions of > or = 65, > or = 70, and > or = 75 years. The rates of enrollment in each age group for each cancer were compared with the corresponding rates in the US cancer population. The age distributions of the US cancer population were derived from the Surveillance, Epidemiology, and End Results Program of the National Cancer Institute for the period 1995 to 1999 based on the 2000 US Census. RESULTS: The proportions of the overall patient populations aged > or = 65, > or = 70, and > or = 75 years were 36%, 20%, and 9% compared with 60%, 46%, and 31%, respectively, in the US cancer population. Statistically significant under-representation of the elderly (P < .001) was noted in registration trials for all cancer treatment except for breast cancer hormonal therapies. Patients aged > or = 70 years accounted for most of the under-representation. CONCLUSION: Elderly were under-represented in the registration trials of new cancer therapies. Various strategies may be needed to evaluate cancer therapies for the elderly in prospective clinical trials and to improve cancer care in the elderly population. PMID- 15542813 TI - DNA methylation and cancer. AB - DNA methylation is an important regulator of gene transcription, and its role in carcinogenesis has been a topic of considerable interest in the last few years. Alterations in DNA methylation are common in a variety of tumors as well as in development. Of all epigenetic modifications, hypermethylation, which represses transcription of the promoter regions of tumor suppressor genes leading to gene silencing, has been most extensively studied. However, global hypomethylation has also been recognized as a cause of oncogenesis. New information concerning the mechanism of methylation and its control has led to the discovery of many regulatory proteins and enzymes. The contribution of dietary folate and methylene terahydrofolate reductase polymorphisms to methylation patterns in normal and cancer tissues is under intense investigation. As methylation occurs early and can be detected in body fluids, it may be of potential use in early detection of tumors and for determining the prognosis. Because DNA methylation is reversible, drugs like 5'-azacytidine, decitabine, and histone deacetylase inhibitors are being used to treat a variety of tumors. Novel demethylating agents such as antisense DNA methyl transferase and small interference RNA are being developed, making the field of DNA methylation wider and more exciting. PMID- 15542814 TI - Challenges in outpatient end-of-life care: wishes to avoid resuscitation. PMID- 15542815 TI - Complications of therapy in cancer patients: Case 1. Paronychia and skin hyperpigmentation induced by gefitinib in advanced non-small-cell lung cancer. PMID- 15542816 TI - Complications of therapy in cancer patients: Case 2. Scrotal ulceration during all-trans-retinoic acid therapy for acute promyelocytic leukemia. PMID- 15542817 TI - Complications of therapy in cancer patients: Case 3. Toxic epidermal necrolysis induced by oral phenobarbital and whole-brain radiotherapy in a breast cancer patient. PMID- 15542818 TI - Mammographic screening is dramatically changing age-incidence data for breast cancer. PMID- 15542819 TI - Imatinib mesylate may improve fasting blood glucose in diabetic Ph+ chronic myelogenous leukemia patients responsive to treatment. PMID- 15542820 TI - Geriatric oncology: a new subspecialty? PMID- 15542821 TI - Interaction between a G-patch protein and a spliceosomal DEXD/H-box ATPase that is critical for splicing. AB - Prp2 is an RNA-dependent ATPase that activates the spliceosome before the first transesterification reaction of pre-mRNA splicing. Prp2 has extensive homology throughout the helicase domain characteristic of DEXD/H-box helicases and a conserved carboxyl-terminal domain also found in the spliceosomal helicases Prp16, Prp22, and Prp43. Despite the extensive homology shared by these helicases, each has a distinct, sequential role in splicing; thus, uncovering the determinants of specificity becomes crucial to the understanding of Prp2 and the other DEAH-splicing helicases. Mutations in an 11-mer near the C-terminal end of Prp2 eliminate its spliceosome binding and splicing activity. Here we show that a helicase-associated protein interacts with this domain and that this interaction contributes to the splicing process. First, a genome-wide yeast two-hybrid screen using Prp2 as bait identified Spp2, which contained a motif with glycine residues found in a number of RNA binding proteins. SPP2 was originally isolated as a genetic suppressor of a prp2 mutant. In a reciprocal screen, Spp2 specifically pulled out the C-terminal half of Prp2. Mutations in the Prp2 C-terminal 11-mer that disrupted function or spliceosome binding also disrupted Spp2 interaction. A screen of randomly mutagenized SPP2 clones identified an Spp2 protein with a mutation in the G patch that could restore interaction with Prp2 and enhanced splicing in a prp2 mutant strain. The study identifies a potential mechanism for Prp2 specificity mediated through a unique interaction with Spp2 and elucidates a role for a helicase-associated protein in the binding of a DEXD/H-box protein to the spliceosome. PMID- 15542822 TI - Evidence for eviction and rapid deposition of histones upon transcriptional elongation by RNA polymerase II. AB - Biochemical experiments indicate that transcriptional elongation by RNA polymerase II (Pol II) is inhibited by nucleosomes and hence requires chromatin modifying activities. Here, we examine the fate of histones upon passage of elongating Pol II in vivo. Histone density throughout the entire Saccharomyces cerevisiae GAL10 coding region is inversely correlated with Pol II association and transcriptional activity, suggesting that the elongating Pol II machinery efficiently evicts core histones from the DNA. Furthermore, new histones appear to be deposited onto DNA less than 1 min after passage of Pol II. Transcription dependent deposition of histones requires the FACT complex that travels with elongating Pol II. Our results suggest that Pol II transcription generates a highly dynamic equilibrium of histone eviction and histone deposition and that there is significant histone exchange throughout most of the yeast genome within a single cell cycle. PMID- 15542823 TI - Ski negatively regulates erythroid differentiation through its interaction with GATA1. AB - The Ski oncoprotein dramatically affects cell growth, differentiation, and/or survival. Recently, Ski was shown to act in distinct signaling pathways including those involving nuclear receptors, transforming growth factor beta, and tumor suppressors. These divergent roles of Ski are probably dependent on Ski's capacity to bind multiple partners with disparate functions. In particular, Ski alters the growth and differentiation program of erythroid progenitor cells, leading to malignant leukemia. However, the mechanism underlying this important effect has remained elusive. Here we show that Ski interacts with GATA1, a transcription factor essential in erythropoiesis. Using a Ski mutant deficient in GATA1 binding, we show that this Ski-GATA1 interaction is critical for Ski's ability to repress GATA1-mediated transcription and block erythroid differentiation. Furthermore, the repression of GATA1-mediated transcription involves Ski's ability to block DNA binding of GATA1. This finding is in marked contrast to those in previous reports on the mechanism of repression by Ski, which have described a model involving the recruitment of corepressors into DNA bound transcription complexes. We propose that Ski cooperates in the process of transformation in erythroid cells by interfering with GATA1 function, thereby contributing to erythroleukemia. PMID- 15542824 TI - A Tel1/MRX-dependent checkpoint inhibits the metaphase-to-anaphase transition after UV irradiation in the absence of Mec1. AB - In Saccharomyces cerevisiae, Mec1/ATR plays a primary role in sensing and transducing checkpoint signals in response to different types of DNA lesions, while the role of the Tel1/ATM kinase in DNA damage checkpoints is not as well defined. We found that UV irradiation in G(1) in the absence of Mec1 activates a Tel1/MRX-dependent checkpoint, which specifically inhibits the metaphase-to anaphase transition. Activation of this checkpoint leads to phosphorylation of the downstream checkpoint kinases Rad53 and Chk1, which are required for Tel1 dependent cell cycle arrest, and their adaptor Rad9. The spindle assembly checkpoint protein Mad2 also partially contributes to the G(2)/M arrest of UV irradiated mec1Delta cells independently of Rad53 phosphorylation and activation. The inability of UV-irradiated mec1Delta cells to undergo anaphase can be relieved by eliminating the anaphase inhibitor Pds1, whose phosphorylation and stabilization in these cells depend on Tel1, suggesting that Pds1 persistence may be responsible for the inability to undergo anaphase. Moreover, while UV irradiation can trigger Mec1-dependent Rad53 phosphorylation and activation in G(1)- and G(2)-arrested cells, Tel1-dependent checkpoint activation requires entry into S phase independently of the cell cycle phase at which cells are UV irradiated, and it is decreased when single-stranded DNA signaling is affected by the rfa1-t11 allele. This indicates that UV-damaged DNA molecules need to undergo structural changes in order to activate the Tel1-dependent checkpoint. Active Clb cyclin-dependent kinase 1 (CDK1) complexes also participate in triggering this checkpoint and are required to maintain both Mec1- and Tel1-dependent Rad53 phosphorylation, suggesting that they may provide critical phosphorylation events in the DNA damage checkpoint cascade. PMID- 15542825 TI - Stimulus-coupled spatial restriction of extracellular signal-regulated kinase 1/2 activity contributes to the specificity of signal-response pathways. AB - Current understanding of cell regulatory systems suggests a diverse array of extracellular stimuli commonly recruit a limited cadre of core signal transduction modules to drive discrete stimulus-specific responses. One such module is the Raf-MEK-extracellular signal-regulated kinase (ERK) kinase cascade. Little information exists about how this pathway can be appropriately coupled to discrete cell biological processes. Contributing factors may include regulation of the duration, amplitude, and/or subcellular compartmentalization of active ERK1/2. To define properties of ERK1/2 that may help mediate stimulus-selective signal propagation, we have examined the dynamic behavior of native ERK1/2 activation at the single-cell level. In primary human cell cultures, ERK1/2 activation is not an all-or-none response. Instead, the amount of active ERK1/2 in individual cells accumulated in proportion to the concentration of external stimulus. The variable degree of ERK1/2 activation correlated well with the degree of ERK1/2 effector activation. Therefore, the relative amplitude of ERK1/2 activation within a cell can be modulated and may contribute to the generation of stimulus-specific biological responses. Importantly, we also found that the capacity of active ERK1/2 to accumulate in the nucleus and drive immediate-early gene expression is dependent upon the nature of the inductive signal, but independent of the amplitude of ERK1/2 activation. Therefore, nuclear accumulation of active ERK1/2 is a discrete regulated step that can direct the function of the kinase in response to specific stimuli. PMID- 15542826 TI - Jumonji represses atrial natriuretic factor gene expression by inhibiting transcriptional activities of cardiac transcription factors. AB - Mice with a homozygous knockout of the jumonji (jmj) gene showed abnormal heart development and defective regulation of cardiac-specific genes, including the atrial natriuretic factor (ANF). ANF is one of the earliest markers of cardiac differentiation and a hallmark for cardiac hypertrophy. Here, we show that JMJ represses ANF gene expression by inhibiting transcriptional activities of Nkx2.5 and GATA4. JMJ represses the Nkx2.5- or GATA4-dependent activation of the reporter genes containing the ANF promoter-enhancer or containing the Nkx2.5 or GATA4-binding consensus sequence. JMJ physically associates with Nkx2.5 and GATA4 in vitro and in vivo as determined by glutathione S-transferase pull-down and immunoprecipitation assays. Using mutational analyses, we mapped the protein protein interaction domains in JMJ, Nkx2.5, and GATA4. We identified two DNA binding sites of JMJ in the ANF enhancer by gel mobility shift assays. However, these JMJ-binding sites do not seem to mediate ANF repression by JMJ. Mutational analysis of JMJ indicates that the protein-protein interaction domain of JMJ mediates the repression of ANF gene expression. Therefore, JMJ may play important roles in the down-regulation of ANF gene expression and in heart development. PMID- 15542827 TI - Translational repression mediates activation of nuclear factor kappa B by phosphorylated translation initiation factor 2. AB - Numerous stressful conditions activate kinases that phosphorylate the alpha subunit of translation initiation factor 2 (eIF2alpha), thus attenuating mRNA translation and activating a gene expression program known as the integrated stress response. It has been noted that conditions associated with eIF2alpha phosphorylation, notably accumulation of unfolded proteins in the endoplasmic reticulum (ER), or ER stress, are also associated with activation of nuclear factor kappa B (NF-kappaB) and that eIF2alpha phosphorylation is required for NF kappaB activation by ER stress. We have used a pharmacologically activable version of pancreatic ER kinase (PERK, an ER stress-responsive eIF2alpha kinase) to uncouple eIF2alpha phosphorylation from stress and found that phosphorylation of eIF2alpha is both necessary and sufficient to activate both NF-kappaB DNA binding and an NF-kappaB reporter gene. eIF2alpha phosphorylation-dependent NF kappaB activation correlated with decreased levels of the inhibitor IkappaBalpha protein. Unlike canonical signaling pathways that promote IkappaBalpha phosphorylation and degradation, eIF2alpha phosphorylation did not increase phosphorylated IkappaBalpha levels or affect the stability of the protein. Pulse chase labeling experiments indicate instead that repression of IkappaBalpha translation plays an important role in NF-kappaB activation in cells experiencing high levels of eIF2alpha phosphorylation. These studies suggest a direct role for eIF2alpha phosphorylation-dependent translational control in activating NF-kappaB during ER stress. PMID- 15542828 TI - G protein-coupled receptor kinase 5 contains a DNA-binding nuclear localization sequence. AB - G protein-coupled receptor kinases (GRKs) mediate desensitization of agonist occupied G protein-coupled receptors (GPCRs). Here we report that GRK5 contains a DNA-binding nuclear localization sequence (NLS) and that its nuclear localization is regulated by GPCR activation, results that suggest potential nuclear functions for GRK5. As assessed by fluorescence confocal microscopy, transfected and endogenous GRK5 is present in the nuclei of HEp2 cells. Mutation of basic residues in the catalytic domain of GRK5 (between amino acids 388 and 395) results in the nuclear exclusion of the mutant enzyme (GRK5(Delta)(NLS)), demonstrating that GRK5 contains a functional NLS. The nuclear localization of GRK5 is subject to dynamic regulation. Calcium ionophore treatment or activation of Gq-coupled muscarinic-M3 receptors promotes the nuclear export of the kinase in a Ca(2+)/calmodulin (Ca(2+)/CaM)-dependent fashion. Ca(2+)/CaM binding to the N-terminal CaM binding site of GRK5 mediates this effect. Furthermore, GRK5, but not GRK5(Delta)(NLS) or GRK2, binds specifically and directly to DNA in vitro. Consistent with their presence in the nuclei of transfected cells, all the GRK4, but not GRK2, subfamily members contain putative NLSs. These results suggest that the GRK4 subfamily of GRKs may play a signaling role in the nucleus and that GRK4 and GRK2 subfamily members perform divergent cellular functions. PMID- 15542829 TI - Yeast chromatin assembly complex 1 protein excludes nonacetylatable forms of histone H4 from chromatin and the nucleus. AB - In yeast, the establishment and maintenance of a transcriptionally silent chromatin state are dependent upon the acetylation state of the N terminus of histone proteins. Histone H4 proteins that contain mutations in N-terminal lysines disrupt heterochromatin and result in yeast that cannot mate. Introduction of a wild-type copy of histone H4 restores mating, despite the presence of the mutant protein, suggesting that mutant H4 protein is either excluded from, or tolerated in, chromatin. To understand how the cell differentiates wild-type histone and mutant histone in which the four N-terminal lysines were replaced with alanine (H4-4A), we analyzed silencing, growth phenotypes, and the histone composition of chromatin in yeast strains coexpressing equal amounts of wild-type and mutant H4 proteins (histone H4 heterozygote). We found that histone H4 heterozygotes have defects in heterochromatin silencing and growth, implying that mutations in H4 are not completely recessive. Nuclear preparations from histone H4 heterozygotes contained less mutant H4 than wild-type H4, consistent with the idea that cells exclude some of the mutant histone. Surprisingly, the N-terminal nuclear localization signal of H4-4A fused to green fluorescent protein was defective in nuclear localization, while a mutant in which the four lysines were replaced with arginine (H4-4R) appeared to have normal nuclear import, implying a role for the charged state of the acetylatable lysines in the nuclear import of histones. The biased partial exclusion of H4-4A was dependent upon Cac1p, the largest subunit of yeast chromatin assembly factor 1 (CAF-1), as well as upon the karyopherin Kap123p, but was independent of Cac2p, another CAF-1 component, and other chromatin assembly proteins (Hir3p, Nap1p, and Asf1p). We conclude that N terminal lysines of histone H4 are important for efficient histone nuclear import. In addition, our data support a model whereby Cac1p and Kap123 cooperate to ensure that only appropriately acetylated histone H4 proteins are imported into the nucleus. PMID- 15542830 TI - Transcription factor binding and induced transcription alter chromosomal c-myc replicator activity. AB - The observation that transcriptionally active genes generally replicate early in S phase and observations of the interaction between transcription factors and replication proteins support the thesis that promoter elements may have a role in DNA replication. To test the relationship between transcription and replication we constructed HeLa cell lines in which inducible green fluorescent protein (GFP) encoding genes replaced the proximal approximately 820-bp promoter region of the c-myc gene. Without the presence of an inducer, basal expression occurred from the GFP gene in either orientation and origin activity was restored to the mutant c-myc replicator. In contrast, replication initiation was repressed upon induction of transcription. When basal or induced transcription complexes were slowed by the presence of alpha-amanitin, origin activity depended on the orientation of the transcription unit. To test mechanistically whether basal transcription or transcription factor binding was sufficient for replication rescue by the uninduced GFP genes, a GAL4p binding cassette was used to replace all regulatory sequences within approximately 1,400 bp 5' to the c-myc gene. In these cells, expression of a CREB-GAL4 fusion protein restored replication origin activity. These results suggest that transcription factor binding can enhance replication origin activity and that high levels of expression or the persistence of transcription complexes can repress it. PMID- 15542831 TI - Diminished S-phase cyclin-dependent kinase function elicits vital Rad53-dependent checkpoint responses in Saccharomyces cerevisiae. AB - Cyclin-dependent kinase (CDK) is required for the initiation of chromosomal DNA replication in eukaryotes. In Saccharomyces cerevisiae, the Clb5 and Clb6 cyclins activate Cdk1 and drive replication origin firing. Deletion of CLB5 reduces initiation of DNA synthesis from late-firing origins. We have examined whether checkpoints are activated by loss of Clb5 function and whether checkpoints are responsible for the DNA replication defects associated with loss of Clb5 function. We present evidence for activation of Rad53 and Ddc2 functions with characteristics suggesting the presence of DNA damage. Deficient late origin firing in clb5Delta cells is not due to checkpoint regulation, but instead, directly reflects the decreased abundance of S-phase CDK, as Clb6 activates late origins when its dosage is increased. Moreover, the viability of clb5Delta cells depends on Rad53. Activation of Rad53 by either Mrc1 or Rad9 contributes to the survival of clb5Delta cells, suggesting that both DNA replication and damage pathways are responsive to the decreased origin usage. These results suggest that reduced origin usage leads to stress or DNA damage at replication forks, necessitating the function of Rad53 in fork stabilization. Consistent with the notion that decreased S-CDK function creates stress at replication forks, deletion of RRM3 helicase, which facilitates replisome progression, greatly diminished the growth of clb5Delta cells. Together, our findings indicate that deregulation of S-CDK function has the potential to exacerbate genomic instability by reducing replication origin usage. PMID- 15542832 TI - Nuclear speckle-associated protein Pnn/DRS binds to the transcriptional corepressor CtBP and relieves CtBP-mediated repression of the E-cadherin gene. AB - Previously, we have shown that pinin/DRS (Pnn), a 140-kDa nuclear and cell adhesion-related phosphoprotein, is involved in the regulation of cell adhesion and modulation of the activity of multiple tumor suppressor genes. In the nucleus Pnn is concentrated in the "nuclear speckles," zones of accumulation of transcriptional and mRNA splicing factors, where Pnn is involved in mRNA processing. Alternatively, other roles of Pnn in gene regulation have not yet been established. By utilizing in vitro pull-down assays, in vivo interaction studies, and immunofluorescence in combination with overexpression and RNA interference experiments, we present evidence that Pnn interacts with the known transcriptional corepressor CtBP1. As a consequence of this interaction Pnn was capable of relieving the CtBP1-mediated repression of E-cadherin promoter activity. Our results suggest that the interaction of Pnn with the corepressor CtBP1 may modulate repression of transcription by CtBP1. This interaction may reflect the existence of coupling factors involved in CtBP-mediated transcriptional regulation and mRNA processing events. PMID- 15542833 TI - Human matrix attachment regions are necessary for the establishment but not the maintenance of transgene insulation in Drosophila melanogaster. AB - Human matrix attachment regions (MARs) can insulate transgene expression from chromosomal position effects in Drosophila melanogaster. To gain insight into the mechanism(s) by which chromosomal insulation occurs, we studied the expression phenotypes of Drosophila transformants expressing mini-white transgenes in which MAR sequences from the human apoB gene were arranged in a variety of ways. In agreement with previous reports, we found that a single copy of the insulating element was not sufficient for position-independent transgene expression; rather, two copies were required. However, the arrangement of the two elements within the transgene was unimportant, since chromosomal insulation was equally apparent when both copies of the insulator were upstream of the mini-white reporter as when the transcription unit was flanked by insulator elements. Moreover, experiments in which apoB 3' MAR sequences were removed from integrated transgenes in vivo by site-specific recombination demonstrated that MAR sequences were required for the establishment but not for the maintenance of chromosomal insulation. These observations are not compatible with the chromosomal loop model in its simplest form. Alternate mechanisms for MAR function in this system are proposed. PMID- 15542834 TI - In vivo analysis of importin alpha proteins reveals cellular proliferation inhibition and substrate specificity. AB - The "classical" nuclear import pathway depends on importins alpha and beta. Humans have only one importin beta, while six alpha importins have been described. Whether or not distinct alpha importins are essential for specific import pathways in living human cells is unclear. We used RNA interference technology to specifically down-regulate the expression of ubiquitously expressed human alpha importins in HeLa cells. Down-regulation of importins alpha3, alpha5, alpha7, and beta strongly inhibited HeLa cell proliferation, while down regulation of importins alpha1 and alpha4 had only a minor effect or no effect. Nucleoplasmin import was not prevented by down-regulation of any alpha importin, indicating that the importin alpha/beta pathway was generally not affected. In contrast, importin alpha3 or alpha5 down-regulation specifically inhibited the nuclear import of the Ran guanine nucleotide exchange factor, RCC1. Coinjection of recombinant alpha importins and RCC1 into down-regulated cells demonstrated that these transport defects were specifically caused by the limited availability of importin alpha3 in both cases. Thus, importin alpha3 is the only alpha importin responsible for the classical nuclear import of RCC1 in living cells. PMID- 15542835 TI - B-cell translocation gene 2 (Btg2) regulates vertebral patterning by modulating bone morphogenetic protein/smad signaling. AB - Btg2 is a primary p53 transcriptional target gene which may function as a coactivator-corepressor and/or an adaptor molecule that modulates the activities of its interacting proteins. We have generated Btg2-null mice to elucidate the in vivo function of Btg2. Btg2-null mice are viable and fertile but exhibit posterior homeotic transformations of the axial vertebrae in a dose-dependent manner. Consistent with its role in vertebral patterning, Btg2 is expressed in the presomitic mesoderm, tail bud, and somites during somitogenesis. We further provide biochemical evidence that Btg2 interacts with bone morphogenetic protein (BMP)-activated Smads and enhances the transcriptional activity of BMP signaling. In view of the genetic evidence that reduced BMP signaling causes posteriorization of the vertebral pattern, we propose that the observed vertebral phenotype in Btg2-null mice is due to attenuated BMP signaling. PMID- 15542836 TI - Multiple, distant Gata2 enhancers specify temporally and tissue-specific patterning in the developing urogenital system. AB - Transcription factor GATA-2 is expressed in a complex temporally and tissue specific pattern within the developing embryo. Loss-of-function studies in the mouse showed that GATA-2 activity is first required during very early hematopoiesis. We subsequently showed that a 271-kbp yeast artificial chromosome (YAC) transgene could fully complement the loss of Gata2 hematopoietic function but that these YAC-rescued Gata2 null mutant mice die perinatally due to defective urogenital development. The rescuing YAC did not display appropriate urogenital expression of Gata2, implying the existence of a urogenital-specific enhancer(s) lying outside the boundaries of this transgene. Here we outline a coupled general strategy for regulatory sequence discovery, linking bioinformatics to functional genomics based on the bacterial artificial chromosome (BAC) libraries used to generate the mouse genome sequence. Exploiting this strategy, we screened >1 Mbp of genomic DNA surrounding Gata2 for urogenital enhancer activity. We found that the spatially and tissue-specific functions for Gata2 in the developing urogenital system are conferred by at least three separate regionally and temporally specific urogenital enhancer elements, two of which reside far 3' to the Gata2 structural gene. Including the additional enhancers that were discovered using this strategy (called BAC trap) extends the functional realm of the Gata2 locus to greater than 1 Mbp. PMID- 15542837 TI - SWAP-70 regulates c-kit-induced mast cell activation, cell-cell adhesion, and migration. AB - SWAP-70, an unusual phosphatidylinositol-3-kinase-dependent protein that interacts with the RhoGTPase Rac, is highly expressed in mast cells. Cultured bone marrow mast cells (BMMC) from SWAP-70(-/-) mice are reduced in FcepsilonRI triggered degranulation. This report describes the hitherto-unknown role of SWAP 70 in c-kit receptor signaling, a key proliferation and differentiation pathway in mast cells. Consistent with the role of Rac in cell motility and regulation of the actin cytoskeleton, mutant cells show abnormal actin rearrangements and are deficient in migration in vitro and in vivo. SWAP-70(-/-) BMMC are impaired in calcium flux, in proper translocation and activity of Akt kinase (required for mast cell activation and survival), and in translocation of Rac1 and Rac2 upon c kit stimulation. Adhesion to fibronectin is reduced, but homotypic cell association induced through c-kit is strongly increased in SWAP-70(-/-) BMMC. Homotypic association requires extracellular Ca(2+) and depends on the integrin alpha(L)beta(2) (LFA-1). ERK is hyperactivated upon c-kit signaling in adherent and dispersed mutant cells. Together, we suggest that SWAP-70 is an important regulator of specific effector pathways in c-kit signaling, including mast cell activation, migration, and cell adhesion. PMID- 15542838 TI - Bcr-Abl-mediated protection from apoptosis downstream of mitochondrial cytochrome c release. AB - Bcr-Abl, activated in chronic myelogenous leukemias, is a potent cell death inhibitor. Previous reports have shown that Bcr-Abl prevents apoptosis through inhibition of mitochondrial cytochrome c release. We report here that Bcr-Abl also inhibits caspase activation after the release of cytochrome c. Bcr-Abl inhibited caspase activation by cytochrome c added to cell-free lysates and prevented apoptosis when cytochrome c was microinjected into intact cells. Bcr Abl acted posttranslationally to prevent the cytochrome c-induced binding of Apaf 1 to procaspase 9. Although Bcr-Abl prevented interaction of endogenous Apaf-1 with the recombinant prodomain of caspase 9, it did not affect the association of endogenous caspase 9 with the isolated Apaf-1 caspase recruitment domain (CARD) or Apaf-1 lacking WD-40 repeats. These data suggest that Apaf-1 recruitment of caspase 9 is faulty in the presence of Bcr-Abl and that cytochrome c/dATP-induced exposure of the Apaf-1 CARD is likely defective. These data provide a novel locus of Bcr-Abl antiapoptotic action and suggest a distinct mechanism of apoptosomal inhibition. PMID- 15542839 TI - Dependence of ORC silencing function on NatA-mediated Nalpha acetylation in Saccharomyces cerevisiae. AB - N(alpha) acetylation is one of the most abundant protein modifications in eukaryotes and is catalyzed by N-terminal acetyltransferases (NATs). NatA, the major NAT in Saccharomyces cerevisiae, consists of the subunits Nat1p, Ard1p, and Nat5p and is necessary for the assembly of repressive chromatin structures. Here, we found that Orc1p, the large subunit of the origin recognition complex (ORC), required NatA acetylation for its role in telomeric silencing. NatA functioned genetically through the ORC binding site of the HMR-E silencer. Furthermore, tethering Orc1p directly to the silencer circumvented the requirement for NatA in silencing. Orc1p was N(alpha) acetylated in vivo by NatA. Mutations that abrogated its ability to be acetylated caused strong telomeric derepression. Thus, N(alpha) acetylation of Orc1p represents a protein modification that modulates chromatin function in S. cerevisiae. Genetic evidence further supported a functional link between NatA and ORC: (i) nat1Delta was synthetically lethal with orc2-1 and (ii) the synthetic lethality between nat1Delta and SUM1-1 required the Orc1 N terminus. We also found Sir3p to be acetylated by NatA. In summary, we propose a model by which N(alpha) acetylation is required for the binding of silencing factors to the N terminus of Orc1p and Sir3p to recruit heterochromatic factors and establish repression. PMID- 15542840 TI - Activation of the DNA damage checkpoint in yeast lacking the histone chaperone anti-silencing function 1. AB - The packaging of the eukaryotic genome into chromatin is likely to be important for the maintenance of genomic integrity. Chromatin structures are assembled onto newly synthesized DNA by the action of chromatin assembly factors, including anti silencing function 1 (ASF1). To investigate the role of chromatin structure in the maintenance of genomic integrity, we examined budding yeast lacking the histone chaperone Asf1p. We found that yeast lacking Asf1p accumulate in metaphase of the cell cycle due to activation of the DNA damage checkpoint. Furthermore, yeast lacking Asf1p are highly sensitive to mutations in DNA polymerase alpha and to DNA replicational stresses. Although yeast lacking Asf1p do complete DNA replication, they have greatly elevated rates of DNA damage occurring during DNA replication, as indicated by spontaneous Ddc2p-green fluorescent protein foci. The presence of elevated levels of spontaneous DNA damage in asf1 mutants is due to increased DNA damage, rather than the failure to repair double-strand DNA breaks, because asf1 mutants are fully functional for double-strand DNA repair. Our data indicate that the altered chromatin structure in asf1 mutants leads to elevated rates of spontaneous recombination, mutation, and DNA damage foci formation arising during DNA replication, which in turn activates cell cycle checkpoints that respond to DNA damage. PMID- 15542841 TI - Proapoptotic function of the MET tyrosine kinase receptor through caspase cleavage. AB - The MET tyrosine kinase, the receptor of hepatocyte growth factor-scatter factor (HGF/SF), is known to be essential for normal development and cell survival. We report that stress stimuli induce the caspase-mediated cleavage of MET in physiological cellular targets, such as epithelial cells, embryonic hepatocytes, and cortical neurons. Cleavage occurs at aspartic residue 1000 within the SVD site of the juxtamembrane region, independently of the crucial docking tyrosine residues Y1001 or Y1347 and Y1354. This cleavage generates an intracellular 40 kDa MET fragment containing the kinase domain. The p40 MET fragment itself causes apoptosis of MDCK epithelial cells and embryonic cortical neurons, whereas its kinase-dead version is impaired in proapoptotic activity. Finally, HGF/SF treatment does not favor MET cleavage and apoptosis, confirming the known survival role of ligand-activated MET. Our results show that stress stimuli convert the MET survival receptor into a proapoptotic factor. PMID- 15542842 TI - Ternary complex factor-serum response factor complex-regulated gene activity is required for cellular proliferation and inhibition of apoptotic cell death. AB - Members of the ternary complex factor (TCF) subfamily of the ETS-domain transcription factors are activated through phosphorylation by mitogen-activated protein kinases (MAPKs) in response to a variety of mitogenic and stress stimuli. The TCFs bind and activate serum response elements (SREs) in the promoters of target genes in a ternary complex with a second transcription factor, serum response factor (SRF). The association of TCFs with SREs within immediate-early gene promoters is suggestive of a role for the ternary TCF-SRF complex in promoting cell cycle entry and proliferation in response to mitogenic signaling. Here we have investigated the downstream gene regulatory and phenotypic effects of inhibiting the activity of genes regulated by TCFs by expressing a dominantly acting repressive form of the TCF, Elk-1. Inhibition of ternary complex activity leads to the downregulation of several immediate-early genes. Furthermore, blocking TCF-mediated gene expression leads to growth arrest and triggers apoptosis. By using mutant Elk-1 alleles, we demonstrated that these effects are via an SRF-dependent mechanism. The antiapoptotic gene Mcl-1 is identified as a key target for the TCF-SRF complex in this system. Thus, our data confirm a role for TCF-SRF-regulated gene activity in regulating proliferation and provide further evidence to indicate a role in protecting cells from apoptotic cell death. PMID- 15542843 TI - Sustained activation of p38 mitogen-activated protein kinase and c-Jun N-terminal kinase pathways by hepatitis B virus X protein mediates apoptosis via induction of Fas/FasL and tumor necrosis factor (TNF) receptor 1/TNF-alpha expression. AB - Activation of the cellular stress pathways (c-Jun N-terminal kinase [JNK] and p38 mitogen-activated protein [MAP] kinase) is linked to apoptosis. However, whether both pathways are required for apoptosis remains unresolved. Hepatitis B virus X protein (pX) activates p38 MAP kinase and JNK pathways and, in response to weak apoptotic signals, sensitizes hepatocytes to apoptosis. Employing hepatocyte cell lines expressing pX, which was regulated by tetracycline, we investigated the mechanism of apoptosis by p38 MAP kinase and JNK pathway activation. Inhibition of the p38 MAP kinase pathway rescues by 80% the initiation of pX-mediated apoptosis, whereas subsequent apoptotic events involve both pathways. pX-mediated activation of p38 MAP kinase and JNK pathways is sustained, inducing the transcription of the death receptor family genes encoding Fas/FasL and tumor necrosis factor receptor 1 (TNFR1)/TNF-alpha and the p53-regulated Bax and Noxa genes. The pX-dependent expression of Fas/FasL and TNFR1/TNF-alpha mediates caspase 8 activation, resulting in Bid cleavage. In turn, activated Bid, acting with pX-induced Bax and Noxa, mediates the mitochondrial release of cytochrome c, resulting in the activation of caspase 9 and apoptosis. Combined antibody neutralization of FasL and TNF-alpha reduces by 70% the initiation of pX-mediated apoptosis. These results support the importance of the pX-dependent activation of both the p38 MAP kinase and JNK pathways in pX-mediated apoptosis and suggest that this mechanism of apoptosis occurs in vivo in response to weak apoptotic signals. PMID- 15542844 TI - p53 Stabilization and accumulation induced by human vaccinia-related kinase 1. AB - Variations in intracellular levels of p53 regulate many cellular functions and determine tumor susceptibility. Major mechanisms modulating p53 levels include phosphorylation and interaction of p53 with specific ubiquitin ligases that promote its degradation. N-terminal phosphorylation regulates the interaction of p53 with several regulatory molecules. Vaccinia-related kinase 1 (VRK1) is the prototype of a new Ser-Thr kinase family in the human kinome. VRK1 is located in the nucleus outside the nucleolus. Overexpression of VRK1 increases the stability of p53 by a posttranslational mechanism leading to its accumulation by a mechanism independent of the Chk2 kinase. Catalytically inactive VRK1 protein (a K179E mutant) does not induce p53 accumulation. VRK1 phosphorylates human p53 in Thr18 and disrupts p53-Mdm2 interaction in vitro, although a significant decrease in p53 ubiquitination by Mdm2 in vivo was not detected. VRK1 kinase does not phosphorylate Mdm2. VRK1-mediated p53 stabilization was also detected in Mdm2(-/ ) cells. VRK1 also has an additive effect with MdmX or p300 to stabilize p53, and p300 coactivation and acetylation of p53 is enhanced by VRK1. The p53 stabilized by VRK1 is transcriptionally active. Suppression of VRK1 expression by specific small interfering RNA provokes several defects in proliferation, situating the protein in the regulation of this process. VRK1 might function as a switch controlling the proteins that interact with p53 and thus modifying its stability and activity. We propose VRK1 as the first step in a new pathway regulating p53 activity during cell proliferation. PMID- 15542845 TI - The mouse genomic instability mutation chaos1 is an allele of Polq that exhibits genetic interaction with Atm. AB - chaos1 (for chromosome aberrations occurring spontaneously 1) is a recessive mutation that was originally identified in a phenotype-based screen for chromosome instability mutants in mice. Mutant animals exhibit significantly higher frequencies of spontaneous and radiation- or mitomycin C-induced micronucleated erythrocytes, indicating a potential defect in homologous recombination or interstrand cross-link repair. The chaos1 allele was genetically associated with a missense mutation in Polq, which encodes DNA polymerase theta;. We demonstrate here that chaos1 is a mutant allele of Polq by using two genetic approaches: chaos1 mutant phenotype correction by a bacterial artificial chromosome carrying wild-type Polq and a failed complementation test between chaos1 and a Polq-disrupted allele generated by gene targeting. To investigate the potential involvement of Polq in DNA double-strand break repair, we introduced chaos1 into an Atm (for ataxia telangiectasia mutated)-deficient background. The majority ( approximately 90%) of double-homozygous mice died during the neonatal period. Surviving double mutants exhibited synergistic phenotypes such as severe growth retardation and enhanced chromosome instability. However, remarkably, double mutants had delayed onset of thymic lymphoma, significantly increasing life span. These data suggest a unique role of Polq in maintaining genomic integrity, which is probably distinctive from the major homologous recombination pathway regulated by ATM. PMID- 15542846 TI - 3'-End polishing of the kinetoplastid spliced leader RNA is performed by SNIP, a 3'-->5' exonuclease with a Motley assortment of small RNA substrates. AB - In all trypanosomatids, trans splicing of the spliced leader (SL) RNA is a required step in the maturation of all nucleus-derived mRNAs. The SL RNA is transcribed with an oligo-U 3' extension that is removed prior to trans splicing. Here we report the identification and characterization of a nonexosomal, 3'-->5' exonuclease required for SL RNA 3'-end formation in Trypanosoma brucei. We named this enzyme SNIP (for snRNA incomplete 3' processing). The central 158-amino-acid domain of SNIP is related to the exonuclease III (ExoIII) domain of the 3'-->5' proofreading epsilon subunit of Escherichia coli DNA polymerase III holoenzyme. SNIP had a preference for oligo(U) 3' extensions in vitro. RNA interference mediated knockdown of SNIP resulted in a growth defect and correlated with the accumulation of one- to two- nucleotide 3' extensions of SL RNA, U2 and U4 snRNAs, a five-nucleotide extension of 5S rRNA, and the destabilization of U3 snoRNA and U2 snRNA. SNIP-green fluorescent protein localized to the nucleoplasm, and substrate SL RNA derived from SNIP knockdown cells showed wild-type cap 4 modification, indicating that SNIP acts on SL RNA after cytosolic trafficking. Since the primary SL RNA transcript was not the accumulating species in SNIP knockdown cells, SL RNA 3'-end formation is a multistep process in which SNIP provides the ultimate 3'-end polishing. We speculate that SNIP is part of an organized nucleoplasmic machinery responsible for processing of SL RNA. PMID- 15542847 TI - An intramolecular association between two domains of the protein kinase Fused is necessary for Hedgehog signaling. AB - The protein kinase Fused (Fu) is an integral member of the Hedgehog (Hh) signaling pathway. Although genetic studies demonstrate that Fu is required for the regulation of the Hh pathway, the mechanistic role that it plays remains largely unknown. Given our difficulty in developing an in vitro kinase assay for Fu, we reasoned that the catalytic activity of Fu might be highly regulated. Several mechanisms are known to regulate protein kinases, including self association in either an intra- or an intermolecular fashion. Here, we provide evidence that Hh regulates Fu through intramolecular association between its kinase domain (DeltaFu) and its carboxyl-terminal domain (Fu-tail). We show that DeltaFu and Fu-tail can interact in trans, with or without the kinesin-related protein Costal 2 (Cos2). However, since the majority of Fu is found associated with Cos2 in vivo, we hypothesized that Fu-tail, which binds Cos2 directly, would be able to tether DeltaFu to Cos2. We demonstrate that DeltaFu colocalizes with Cos2 in the presence of Fu-tail and that this colocalization occurs on a subset of membrane vesicles previously characterized to be important for Hh signal transduction. Additionally, expression of Fu-tail in fu mutant flies that normally express only the kinase domain rescues the fu wing phenotype. Therefore, reestablishing the association between these two domains of Fu in trans is sufficient to restore Hh signal transduction in vivo. In such a manner we validate our hypothesis, demonstrating that Fu self-associates and is functional in an Hh-dependent manner. Our results here enhance our understanding of one of the least characterized, yet critical, components of Hh signal transduction. PMID- 15542848 TI - Genetic interaction between Rb and K-ras in the control of differentiation and tumor suppression. AB - Although the retinoblastoma protein (pRb) has been implicated in the processes of cellular differentiation, there is no compelling genetic or in vivo evidence that such activities contribute to pRb-mediated tumor suppression. Motivated by cell culture studies suggesting that Ras is a downstream effector of pRb in the control of differentiation, we have examined the tumor and developmental phenotypes of Rb and K-ras double-knockout mice. We find that heterozygosity for K-ras (i) rescued a unique subset of developmental defects that characterize Rb deficient embryos by affecting differentiation but not proliferation and (ii) significantly enhanced the degree of differentiation of pituitary adenocarcinomas arising in Rb heterozygotes, leading to their prolonged survival. These observations suggest that Rb and K-ras function together in vivo, in the contexts of both embryonic and tumor development, and that the ability to affect differentiation is a major facet of the tumor suppressor function of pRb. PMID- 15542849 TI - Stage-specific repression by the EKLF transcriptional activator. AB - Dynamic changes in transcription factor function can be mediated by switching its interaction with coactivators and corepressors. Erythroid Kruppel-like factor (EKLF) is an erythroid cell-specific transcription factor that plays a critical role in beta-globin gene activation via its interactions with CBP/p300 and SWI/SNF proteins. Unexpectedly, it also interacts with Sin3A and histone deacetylase 1 (HDAC1) corepressors via its zinc finger domain. We now find that selected point mutants can uncouple activation and repression and that an intact finger structure is not required for interactions with Sin3A/HDAC1 or for transrepression. Most intriguingly, EKLF repression exhibits stage specificity, with reversible EKLF-Sin3A interactions playing a key role in this process. Finally, we have located a key lysine residue that is both a substrate for CBP acetylation and required for Sin3A interaction. These data suggest a model whereby the stage of the erythroid cell alters the acetylation status of EKLF and plays a critical role in directing its coactivator-corepressor interactions and downstream transcriptional effects. PMID- 15542850 TI - Partial cleavage of RasGAP by caspases is required for cell survival in mild stress conditions. AB - Tight control of apoptosis is required for proper development and maintenance of homeostasis in multicellular organisms. Cells can protect themselves from potentially lethal stimuli by expressing antiapoptotic factors, such as inhibitors of apoptosis, FLICE (caspase 8)-inhibitory proteins, and members of the Bcl2 family. Here, we describe a mechanism that allows cells to survive once executioner caspases have been activated. This mechanism relies on the partial cleavage of RasGAP by caspase 3 into an amino-terminal fragment called fragment N. Generation of this fragment leads to the activation of the antiapoptotic Akt kinase, preventing further amplification of caspase activity. Partial cleavage of RasGAP is required for cell survival under stress conditions because cells expressing an uncleavable RasGAP mutant cannot activate Akt, cannot prevent amplification of caspase 3 activity, and eventually undergo apoptosis. Executioner caspases therefore control the extent of their own activation by a feedback regulatory mechanism initiated by the partial cleavage of RasGAP that is crucial for cell survival under adverse conditions. PMID- 15542851 TI - A mammalian ortholog of Saccharomyces cerevisiae Vac14 that associates with and up-regulates PIKfyve phosphoinositide 5-kinase activity. AB - Multivesicular body morphology and size are controlled in part by PtdIns(3,5)P(2), produced in mammalian cells by PIKfyve-directed phosphorylation of PtdIns(3)P. Here we identify human Vac14 (hVac14), an evolutionarily conserved protein, present in all eukaryotes but studied principally in yeast thus far, as a novel positive regulator of PIKfyve enzymatic activity. In mammalian cells and tissues, Vac14 is a low-abundance 82-kDa protein, but its endogenous levels could be up-regulated upon ectopic expression of hVac14. PIKfyve and hVac14 largely cofractionated, populated similar intracellular locales, and physically associated. A small-interfering RNA-directed gene-silencing approach to selectively eliminate endogenous hVac14 rendered HEK293 cells susceptible to morphological alterations similar to those observed upon expression of PIKfyve mutants deficient in PtdIns(3,5)P(2) production. Largely decreased in vitro PIKfyve kinase activity and unaltered PIKfyve protein levels were detected under these conditions. Conversely, ectopic expression of hVac14 increased the intrinsic PIKfyve lipid kinase activity. Concordantly, intracellular PtdIns(3)P to-PtdIns(3,5)P(2) conversion was perturbed by hVac14 depletion and was elevated upon ectopic expression of hVac14. These data demonstrate a major role of the PIKfyve-associated hVac14 protein in activating PIKfyve and thereby regulating PtdIns(3,5)P(2) synthesis and endomembrane homeostasis in mammalian cells. PMID- 15542852 TI - Deficiency in SNM1 abolishes an early mitotic checkpoint induced by spindle stress. AB - Spindle poisons represent an important class of anticancer drugs that act by interfering with microtubule polymerization and dynamics and thereby induce mitotic checkpoints and apoptosis. Here we show that mammalian SNM1 functions in an early mitotic stress checkpoint that is distinct from the well-characterized spindle checkpoint that regulates the metaphase-to-anaphase transition. Specifically, we found that compared to wild-type cells, Snm1-deficient mouse embryonic fibroblasts exposed to spindle poisons exhibited elevated levels of micronucleus formation, decreased mitotic delay, a failure to arrest in mitosis prior to chromosome condensation, supernumerary centrosomes, and decreased viability. In addition, we show that both Snm1 and 53BP1, previously shown to interact, coimmunoprecipitate with components of the anaphase-promoting complex (APC)/cyclosome. These findings suggest that Snm1 is a component of a mitotic stress checkpoint that negatively targets the APC prior to chromosome condensation. PMID- 15542853 TI - Disruption of PLZP in mice leads to increased T-lymphocyte proliferation, cytokine production, and altered hematopoietic stem cell homeostasis. AB - Deregulated function of members of the POK (POZ and Kruppel) family of transcriptional repressors, such as promyelocytic leukemia zinc finger (PLZF) and B-cell lymphoma 6 (BCL-6), plays a critical role in the pathogenesis of acute promyelocytic leukemia (APL) and non-Hodgkin's lymphoma, respectively. PLZP, also known as TZFP, FAZF, or ROG, is a novel POK protein that displays strong homology with PLZF and has been implicated in the pathogenesis of the cancer-predisposing syndrome, Fanconi's anemia, and of APL, in view of its ability to heterodimerize with the FANC-C and PLZF proteins, respectively. Here we report the generation and characterization of mice in which we have specifically inactivated the PLZP gene through in-frame insertion of a lacZ reporter and without perturbing the expression of the neighboring MLL2 gene. We show that PLZP-deficient mice display defects in cell cycle control and cytokine production in the T-cell compartment. Importantly, PLZP inactivation perturbs the homeostasis of the hematopoietic stem and/or progenitor cell. On the basis of our data, a deregulation of PLZP function in Fanconi's anemia and APL may affect the biology of the hematopoietic stem cell, in turn contributing to the pathogenesis of these disorders. PMID- 15542854 TI - Binding to nonmethylated CpG DNA is essential for target recognition, transactivation, and myeloid transformation by an MLL oncoprotein. AB - The MLL gene is a frequent target for leukemia-associated chromosomal translocations that generate dominant-acting chimeric oncoproteins. These invariably contain the amino-terminal 1,400 residues of MLL fused with one of a variety of over 30 distinct nuclear or cytoplasmic partner proteins. Despite the consistent inclusion of the MLL amino-terminal region in leukemia oncoproteins, little is known regarding its molecular contributions to MLL-dependent oncogenesis. Using high-resolution mutagenesis, we identified three MLL domains that are essential for in vitro myeloid transformation via mechanisms that do not compromise subnuclear localization. These include the CXXC/Basic domain and two novel domains of unknown function. Point mutations in the CXXC domain that eliminate myeloid transformation by an MLL fusion protein also abolished recognition and binding of nonmethylated CpG DNA sites in vitro and transactivation in vivo. Our results define a critical role for the CXXC DNA binding domain in MLL-associated oncogenesis, most likely via epigenetic recognition of CpG DNA sites within the regulatory elements of target genes. PMID- 15542855 TI - Yeast shuttling SR proteins Npl3p, Gbp2p, and Hrb1p are part of the translating mRNPs, and Npl3p can function as a translational repressor. AB - A major challenge in current molecular biology is to understand how sequential steps in gene expression are coupled. Recently, much attention has been focused on the linkage of transcription, processing, and mRNA export. Here we describe the cytoplasmic rearrangement for shuttling mRNA binding proteins in Saccharomyces cerevisiae during translation. While the bulk of Hrp1p, Nab2p, or Mex67p is not associated with polysome containing mRNAs, significant amounts of the serine/arginine (SR)-type shuttling mRNA binding proteins Npl3p, Gbp2p, and Hrb1p remain associated with the mRNA-protein complex during translation. Interestingly, a prolonged association of Npl3p with polysome containing mRNAs results in translational defects, indicating that Npl3p can function as a negative translational regulator. Consistent with this idea, a mutation in NPL3 that slows down translation suppresses growth defects caused by the presence of translation inhibitors or a mutation in eIF5A. Moreover, using sucrose density gradient analysis, we provide evidence that the import receptor Mtr10p, but not the SR protein kinase Sky1p, is involved in the timely regulated release of Npl3p from polysome-associated mRNAs. Together, these data shed light onto the transformation of an exporting to a translating mRNP. PMID- 15542856 TI - Sox7 plays crucial roles in parietal endoderm differentiation in F9 embryonal carcinoma cells through regulating Gata-4 and Gata-6 expression. AB - During early rodent development, the parietal endoderm appears from an inner cell mass and produces large amounts of basement membrane components, such as laminin 1 and collagen IV. To elucidate the regulatory network for gene expression during these procedures, we constructed a series of short interfering RNA expression vectors targeted to various transcription factors, transfected them into F9 embryonal carcinoma cells, and evaluated the effects of the gene silencing on the induction of parietal endoderm differentiation and basement membrane component production by treating F9 cells with all trans-retinoic acid and dibutyryl cyclic AMP. Among the transcription factors tested, silencing of Sox7 or combined silencing of Gata-4 and Gata-6 resulted in suppression of cell shape changes and laminin-1 production, which are the hallmarks of parietal endoderm differentiation. In cells silenced for Sox7, induction of Gata-4 and Gata-6 by retinoic acid and cyclic AMP treatment was inhibited, while induction of Sox7 was not affected in cells silenced for Gata-4 and Gata-6, indicating that Sox7 is an upstream regulatory factor for these Gata factors. Nevertheless, silencing of Sox7 did not totally cancel the action of retinoic acid, since upregulation of coup-tf2, keratin 19, and retinoic acid receptor beta2 was not abolished in Sox7 silenced F9 cells. Although overexpression of Sox7 alone was insufficient to induce parietal endoderm differentiation, overexpression of Gata-4 or Gata-6 in Sox7-silenced F9 cells restored the differentiation into parietal endoderm. Sox7 is therefore required for the induction of Gata-4 and Gata-6, and the interplay among these transcription factors plays a crucial role in parietal endoderm differentiation. PMID- 15542857 TI - Analysis of the localization and topology of nurim, a polytopic protein tightly associated with the inner nuclear membrane. AB - Nurim is an inner nuclear membrane (INM) protein that was first isolated in a visual screen for nuclear envelope-localizing proteins. Nurim lacks an N-terminal domain characteristic of other INM proteins examined to date and may represent a class of proteins that localize to the INM by a distinct mechanism. To further characterize this protein, we constructed nurim-green fluorescent protein fusions and analyzed aspects of localization, biochemistry, and membrane topology. Results from immunoprobing and protease protection assays together with other analyses indicate that nurim (total length of 262 residues) is a six transmembrane-spanning protein and contains a hairpin turn in its C-terminal transmembrane domain, resulting in the N and C termini residing on the same side of the membrane. A loop region between the fourth and fifth transmembrane domains is exposed toward the nucleoplasm and contains a region accessible for site specific endoproteinase cleavage. In biochemical fractionation, nurim remained extremely tightly bound to nuclear fractions and was released in significant quantities only in the presence of 4 m urea. Under conditions in which nuclear lamins were completely extracted, a significant population of nurim remained resistant to solubilization. This tight binding requires the C-terminal region of the protein. DNase treatment only marginally influenced its retention characteristics in nuclei. Results from consideration of sequence alignments and identification of specific topological features of nurim indicate that it may possess enzymic function. These results are discussed with reference to the retention mechanism and possible nuclear function of nurim. PMID- 15542858 TI - Identification of the structural and functional domains of the large serine recombinase TnpX from Clostridium perfringens. AB - Members of the large serine resolvase family of site-specific recombinases are responsible for the movement of several mobile genetic elements; however, little is known regarding the structure or function of these proteins. TnpX is a serine recombinase that is responsible for the movement of the chloramphenicol resistance elements of the Tn4451/3 family. We have shown that TnpX binds differentially to its transposon and target sites, suggesting that resolvase-like excision and insertion were two distinct processes. To analyze the structural and functional domains of TnpX and, more specifically, to define the domains involved in protein-DNA and protein-protein interactions, we conducted limited proteolysis studies on the wild-type dimeric TnpX(1-707) protein and its functional truncation mutant, TnpX(1-597). The results showed that TnpX was organized into three major domains: domain I (amino acids (aa) 1-170), which included the resolvase catalytic domain; domain II (aa 170-266); and domain III (aa 267-707), which contained the dimerization region and two separate regions involved in binding to the DNA target. A small polypeptide (aa 533-587) was shown to bind specifically to the TnpX binding sites providing further evidence that it was the primary DNA binding region. In addition, a previously unidentified DNA binding site was shown to be located between residues 583 and 707. Finally, the DNA binding and multerimization but not the catalytic functions of TnpX could be reconstituted by recombining separate polypeptides that contain the N- and C terminal regions of the protein. These data provide evidence that TnpX has separate catalytic, DNA binding, and multimerization domains. PMID- 15542859 TI - RNA structural rearrangement via unwinding and annealing by the cyanobacterial RNA helicase, CrhR. AB - Rearrangement of RNA secondary structure is crucial for numerous biological processes. RNA helicases participate in these rearrangements through the unwinding of duplex RNA. We report here that the redox-regulated cyanobacterial RNA helicase, CrhR, is a bona fide RNA helicase possessing both RNA-stimulated ATPase and bidirectional ATP-stimulated RNA helicase activity. The processivity of the unwinding reaction appears to be low, because RNA substrates containing duplex regions of 41 bp are not unwound. CrhR also catalyzes the annealing of complementary RNA into intermolecular duplexes. Uniquely and in contrast to other proteins that perform annealing, the CrhR-catalyzed reactions require ATP hydrolysis. Through a combination of the unwinding and annealing activities, CrhR also catalyzes RNA strand exchange resulting in the formation of RNA secondary structures that are too stable to be resolved by helicase activity. RNA strand exchange most probably occurs through the CrhR-dependent formation and resolution of an RNA branch migration structure. Demonstration that another cyanobacterial RNA helicase, CrhC, does not catalyze annealing indicates that this activity is not a general biochemical characteristic of RNA helicases. Biochemically, CrhR resembles RecA and related proteins that catalyze strand exchange and branch migration on DNA substrates, a characteristic that is reflected in the recently reported structural similarities between these proteins. The data indicate the potential for CrhR to catalyze dynamic RNA secondary structure rearrangements through a combination of RNA helicase and annealing activities. PMID- 15542860 TI - Role of H+-ATPase-mediated acidification in sorting and release of the regulated secretory protein chromogranin A: evidence for a vesiculogenic function. AB - The constitutive and regulated secretory pathways represent the classical routes for secretion of proteins from neuroendocrine cells. Selective aggregation of secretory granule constituents in an acidic, bivalent cation-rich environment is considered to be a prerequisite for sorting to the regulated secretory pathway. The effect of selective vacuolar H+-ATPase (V-ATPase) inhibitor bafilomycin A1 on the pH gradient along the secretory pathway was used here to study the role of acidification on the trafficking of the regulated secretory protein chromogranin A (CgA) in PC12 cells. Sorting of CgA was assessed by three-dimensional deconvolution microscopy, subcellular fractionation, and secretagogue-stimulated release, examining a series of full-length or truncated domains of human CgA (CgA (1-115), CgA-(233-439)) fused to either green fluorescent protein or to a novel form of secreted embryonic alkaline phosphatase (EAP). We show that a full-length CgA/EAP chimera is sorted to chromaffin granules for exocytosis. Inhibition of V ATPase by bafilomycin A1 markedly reduced the secretagogue-stimulated release of CgA-EAP by perturbing sorting of the chimera (at the trans-Golgi network or immature secretory granule) rather than the late steps of exocytosis. The effect of bafilomycin A1 on CgA secretion depends on a sorting determinant located within the amino terminus (CgA-(1-115)) but not the C-terminal region of the granin. Moreover, examination of chromaffin granule abundance in PC12 cells exposed to bafilomycin A1 reveals a substantial decrease in the number of dense core vesicles. We propose that a V-ATPase-mediated pH gradient in the secretory pathway is an important factor for the formation of dense-core granules by regulating the ability of CgA to form aggregates, a crucial step that may underlie the granulogenic function of the protein. PMID- 15542861 TI - Biochemical and NMR mapping of the interface between CREB-binding protein and ligand binding domains of nuclear receptor: beyond the LXXLL motif. AB - CBP, cAMP-response element-binding protein (CREB)-binding protein, plays an important role as a general cointegrator of various signaling pathways and interacts with a large number of transcription factors. Interactions of CBP with ligand binding domains (LBDs) of nuclear receptors are mediated by LXXLL motifs, as are those of p160 proteins, although the number, distribution, and precise sequences of the motifs differ. We used a large N-terminal fragment of murine CBP to map by biochemical methods and NMR spectroscopy the interaction domain of CBP with the LBDs of several nuclear receptors. We show that distinct zones of that fragment are involved in the interactions: a 20-residue segment containing the LXXLL motif (residues 61-80) is implicated in the interaction with all three domains tested (peroxisome proliferator-activated receptor gamma-LBD, retinoid X receptor alpha-LBD, and estrogen-related receptor gamma-LBD), whereas a second N terminal well conserved block of around 25 residues centered on a consensus L(40)PDEL(44) motif constitutes a secondary motif of interaction with peroxisome proliferator-activated receptor gamma-LBD. Sequence analysis reveals that both zones are well conserved in all vertebrate p300/CBP proteins, suggesting their functional importance. Interactions of p300/CBP coactivators with the LBDs of nuclear receptors are not limited to the canonical LXXLL motifs, involving both a longer contiguous segment around the motif and, for certain domains, an additional zone. PMID- 15542863 TI - Effects of unpaired nucleotides within HIV-1 genomic secondary structures on pausing and strand transfer. AB - Reverse transcriptase-mediated RNA displacement synthesis is required for DNA polymerization through the base-paired stem portions of secondary structures present in retroviral genomes. These regions of RNA duplex often possess single unpaired nucleotides, or "bulges," that disrupt contiguous base pairing. By using well defined secondary structures from the human immunodeficiency virus, type 1 (HIV-1), genome, we demonstrate that removal of these bulges either by deletion or by introducing a complementary base on the opposing strand results in increased pausing at specific positions within the RNA duplex. We also show that the HIV-1 nucleocapsid protein can increase synthesis through the pause sites but not as efficiently as when a bulge residue is present. Finally, we demonstrate that removing a bulge increases the proportion of strand transfer events to an acceptor template that occur prior to complete replication of a donor template secondary structure. Together our data suggest a role for bulge nucleotides in enhancing synthesis through stable secondary structures and reducing strand transfer. PMID- 15542864 TI - A proteomic strategy for gaining insights into protein sumoylation in yeast. AB - Sumoylation represents a vital post-translational modification that pervades numerous aspects of cell biology, including protein targeting, transcriptional regulation, signal transduction, and cell division. However, despite its broad reaching effects, most biological outcomes of protein sumoylation remain poorly understood. In an effort to provide further insight into this complex process, a proteomics approach was undertaken to identify the targets of sumoylation en mass. Specifically, SUMO-conjugated proteins were isolated by a double-affinity purification procedure from a Saccharomyces cerevisiae strain engineered to express tagged SUMO. The components of the isolated protein mixture were then identified by subsequent LC-MS/MS analysis using an LTQ FT mass spectrometer. In this manner, 159 candidate sumoylated proteins were identified by two or more peptides. Furthermore, the high accuracy of the instrument, combined with stringent search criteria, enabled the identification of an additional 92 putative candidates by only one peptide. The validity of this proteomics approach was confirmed by performing subsequent Western blot experiments for numerous proteins and determining the actual sumoylation sites for several other substrates. These data combine with recent works to further our understanding of the breadth and impact of protein sumoylation in a diverse array of biological processes. PMID- 15542865 TI - Hendrikje van Andel, bristlecone pines, and why we die. PMID- 15542866 TI - Place, time and certified cause of death in people who die after hospital admission for myocardial infarction or stroke. AB - BACKGROUND: Two areas of uncertainty about routine statistics for mortality after hospital admission for myocardial infarction (MI) or stroke are i) whether most deaths occur in the admission episode itself rather than after discharge, and ii) whether most deaths are certified on death certificates as, respectively, MI or stroke. METHODS: Use of linked hospital and mortality statistics to analyse the time, place and certified cause of death in people aged 35-74 after admission for MI or stroke. RESULTS: Of 7,964 deaths within a year of admission for MI, 5,686 (71.4%) occurred within 30 days of admission. Of these, 4,856 (85.4%) occurred during the initial hospital admission. Of 7,070 deaths within a year of admission for stroke, 4,905 (69.4%) were within 30 days, and 4,509 (91.9%) of these occurred during the initial admission. As expected, deaths at longer intervals than 30 days occurred mainly after discharge. Of deaths within 30 days of MI and stroke, 85.2% and 80.0%, respectively, were certified with MI or stroke as the underlying cause of death. CONCLUSION: In-hospital death rates alone, calculated without record linkage to death certificates, would have identified most deaths that occurred within 30 days of admission. Nonetheless, linkage added to completeness of ascertainment even within this time period. Data without linkage are unreliable in identifying deaths at longer time intervals. Routine mortality statistics for MI and stroke, as the underlying cause, reliably included most deaths that occurred within 30 days of admission for each respective disease. PMID- 15542867 TI - Risk factors for asymptomatic abdominal aortic aneurysm: systematic review and meta-analysis of population-based screening studies. AB - BACKGROUND: The incidence of and mortality from ruptured abdominal aortic aneurysm (AAA) is increasing. There is uncertainty regarding the indicators which could be used to identify groups at high risk. This issue has been addressed in a systematic review of population-based screening studies. METHODS: MEDLINE and EMBASE were searched, reference lists scanned and manual searches made of eight journals. The search was restricted to four languages (English, German, French and Italian). Population-based studies investigating risk factors associated with screening-detected AAA were included. The following risk factors were considered: sex, smoking, hypertension, diabetes, a history myocardial infarction, and peripheral vascular disease. RESULTS: Fourteen cross-sectional studies met our inclusion criteria. Most studies screened people aged 60 years or older. The prevalence of AAA ranged from 4.1% to 14.2% in men and from 0.35% to 6.2% in women. Male sex showed a strong association with AAA (OR 5.69), whereas smoking (OR 2.41), a history of myocardial infarction (OR 2.28) or peripheral vascular disease (OR 2.50) showed moderate associations. Hypertension was only weakly associated with AAA (OR 1.33) and no association was evident with diabetes (OR 1.02). CONCLUSIONS: The efficacy of screening men aged 60 years or older and women of the same age who smoke or have a history of peripheral or coronary artery disease should be evaluated in randomized controlled trials. PMID- 15542868 TI - Return to work after first myocardial infarction in 1991-1996 in Finland. AB - BACKGROUND: A substantial number of myocardial infarctions (MI) occur at working age. It is, however, insufficiently well known how many of these patients return to work after their MI. METHODS: Sources of information were the Hospital Discharge Register, the Causes of Death Register and the registers for social security benefits. Availability for the labour market was used as the return to work criterion. Altogether 10,244 persons (8,733 men, 1,511 women) aged 35-59 years had their first MI or coronary death during 1991-1994 in Finland. Persons who survived for 28 days and were not on pension at the time of MI were included in a two-year follow-up. RESULTS: Twenty-nine per cent of patients were already pensioned at the time of their first MI. Of the patients not pensioned at the time of their MI, 4,929 were alive two years after the event. Of them, 38% of men and 40% of women received disability pension, 3% of both genders were on sick leave and 1% of both genders were on unemployment pension. The remainder, 58% of men and 56% of women, did not receive any of these benefits, thus, being available to the labour force. CONCLUSIONS: Nearly one-third of persons having their first MI at working age were already out of the labour force at the time of their MI. Of those who were not pensioned and who survived the event, slightly more than half were available to the labour market two years later. PMID- 15542869 TI - Parental economic stress: evidence of an overlooked public health risk among Swedish families. AB - BACKGROUND: Studies have demonstrated that when parents shoulder considerable financial responsibilities, adverse health outcomes may occur. The present study assesses the association between economic stress and self-rated health in a sample of Swedish parents, and especially how this relation is affected by foreign origin and employment status. METHODS: A questionnaire was sent to a random sample of 5,600 individuals between the ages of 21 and 81 in Malmo, Sweden. The total response rate was 69%. Among the respondents, 824 were parents having at least one child living at home. The main exposures were such sociodemographic variables as country of origin and employment status, and economic stress. The outcome variable was self-rated health. RESULTS: Of the parents in the study, the 34.7% coded as exposed to economic stress showed a significantly increased odds ratio for poor self-rated health (OR=3.12, 95% CI: 2.01-4.84) adjusted for age and sex. After controlling for foreign origin and unemployment, the odds ratio remained statistically significant regarding exposure to economic stress (OR=1.94; 1.16-3.23). In the multivariate model, foreign origin and unemployment were also strongly associated with poor self rated health (OR=1.78, 95% CI: 1.12-2.88; OR=1.67, 95% CI: 1.01-2.75, respectively). The adjusted population-attributable risk for poor self-rated health was estimated to be 27.4% for economic stress, 26.6% for foreign origin, and 16.7% for unemployment. CONCLUSIONS: Parental economic stress was associated with low self-rated health to a statistically significant degree, even when accounting for employment status and foreign origin. It, therefore, deserves to be seriously considered as an potential public health risk factor among Swedish families. PMID- 15542870 TI - Prevalence of hypertension, hypercholesterolemia, smoking and overweight in older Belgian adolescents. AB - BACKGROUND: Risk factors in young adulthood may persist into later life and eventually lead to cardiovascular (CV) disease. The purpose of this study was to describe the prevalence of hypertension and other established CV risk factors in older adolescents. METHODS: Study nurses examined 120 girls and 80 boys with a mean age of 17.4 (SD, 0.8; range 15.8-19.6 years). After 5 minutes of sitting rest, resting blood pressure (BP) of the participants was measured in triplicate by sphygmomanometry. Body height and weight were measured and body mass index (BMI) was calculated. Serum lipids, smoking and drinking status were assessed. Hypertension was diagnosed according to the criteria specified in the 1996 update of the Task Force on BP treatment in Children and Adolescents (Pediatrics 1996;98:649-58). RESULTS: Almost 25% of the subjects currently smoked with a median daily consumption of 6 (Interquartile 4-9) and 11 (6-16) cigarettes in girls and boys, respectively. Fourty-nine (41%) girls used oral contraceptives. Systolic (118 versus 109 mm Hg, p<0.001) and diastolic BPs (70.5 versus 68.0 mm Hg, p=0.033) were significantly higher in boys than in girls. Two (2%) girls and four (5%) boys had systolic hypertension; diastolic hypertension was found in six (5%) girls and one (1%) boy. Twelve (10%) girls and six (8%) boys were overweight (>25.0 kg/m2). Twenty (17%) girls and seven (9%) boys had hypercholesterolemia (total serum cholesterol > or =5.2 mmol/L). Overall, 50 (42%) girls and 31 (39%) boys had at least one CV risk factor, 12 (10%) girls and four (5%) boys had two risk factors, and three (3%) girls and one (1%) boy had more than two risk factors. CONCLUSIONS: In 17-year-olds living in an affluent society the prevalence of CV risk factors was high. These findings underscore the importance of health education and prevention at this age. PMID- 15542871 TI - Are parents aware of their schoolchildren's headaches? AB - BACKGROUND: The objective of this study is to determine parents' awareness of their children's headaches and to evaluate some of the factors that affect this awareness. METHODS: The subjects of the study are 2601 children who were diagnosed with headache. Data on the children and the parents was collected using a detailed data form. The diagnosis of headache in children was made on the basis of the criteria of the International Headache Society (IHS). If the parents of a child diagnosed with headache reported that their child had headache, the parent was evaluated to be aware of his/her child's headache. In the statistical analyses, chi-square and binary logistic regression were used. RESULTS: Almost 74% of parents were aware of their children's headache. It was found that migraine type headache, female gender, being the first child of the family, travel sickness of children, the presence of headache history in one of the family members; the number of family members and mother's age are factors that affect the awareness level of parents. It was also revealed that parents who do not work outside are more aware of their children's headache and that educational and financial status do not have any effect on the degree of awareness. CONCLUSIONS: In a city like Mersin, which is economically well developed when compared with the rest of the country, one quarter of the parents are not aware of their children's headache. PMID- 15542872 TI - Differences in older adults' use of primary and specialist care services in two Nordic countries. AB - BACKGROUND: The aim of the study was to compare elderly persons' self-reported use of physician services and associated sociodemographic factors and self-rated health in two Nordic countries with different health care systems, Finland and Norway. METHODS: Population based, cross-sectional surveys conducted in Norway (1995-97) and in Finland (1997) were employed. In the Norwegian data a total of 7,919 individuals, and in the Finnish data 1, 500 individuals, aged 65-74 years old were included in the samples. The outcome variables were having visited a general practitioner, a specialist or both during the past 12 months. Associations between utilization of physician's services and sociodemographic factors and self-rated health were analysed by multiple logistic regression. RESULTS: Approximately the same proportion of elderly in Norway and Finland reported having visited a physician during the previous 12 months. Finnish elderly more often visited a specialist compared to Norwegians. Self-rated health was strongly associated with visits to a specialist in both countries and to a GP in Norway. In Finland visits to a GP were only weakly connected with self-rated health. The use of specialist services increased with increasing education in both countries and in Finland the association was steeper than in Norway. Marital status was not consistently associated with visiting a physician. CONCLUSIONS: Higher rates of specialist care among the elderly in Finland may indicate a more efficient gate-keeping role among Norwegian general practitioners or inducement caused by two overlapping service sectors. Inconsistent associations between utilization and health variables may be due to cultural differences. PMID- 15542873 TI - Birthweight differences among infants of North African immigrants and Belgians in Belgium. AB - BACKGROUND: Infants of North African immigrants are reported to have higher birthweights than their Belgian counterparts. It is unclear what mechanism contributes to this difference. METHODS: Analyses were based on a hospital-based cohort of 1,162 women. RESULTS: Infants of North African immigrants were less likely to be born preterm, compared to infants of Belgian women. After adjusting for sociodemographic and maternal factors, the estimated difference in mean birthweight was 74 g (p=0.05). When limited to term births, this difference was 28 g (p=0.42). CONCLUSION: The difference in mean birthweight between North African and Belgian infants was explained by differences in preterm birth and other risk factors. PMID- 15542874 TI - Multiethnic differences in smoking in Israel: pooled analysis from three national surveys. AB - BACKGROUND: Smoking is a major public health risk and information regarding high risk groups is needed to plan, implement and evaluate interventions aimed at lowering the number of smokers. METHODS: During the years 1999-2001 data was collected regarding the smoking behaviour of the Israeli population in three national surveys. All three surveys included ages 25-64 and two included also ages 21-25 and over 64. RESULTS: Smoking was associated with age, sex, ethnicity, education and religiosity after adjusting for the various demographic characteristics of the survey population. The prevalence of smoking among Arab and immigrant men from the former Soviet Union is higher than among Jewish men. Among women the opposite association exists. The older, religious and more educated reported smoking less frequently. In the Jewish population respondents, defining themselves as secular, reported higher rates of smoking. Only in women was marital status associated with smoking. A few specific high-risk groups for smoking can be identified such as young, less educated men, Arab men, single Jewish women and young immigrant men and women. CONCLUSIONS: The three ethnic groups residing in Israel differ in the prevalence of smoking; each has a distinct pattern of smoking, positioning them at different stages within the conceptual framework of the larger smoking pandemic. Ethnicity, religiosity, age and education are associated with smoking in both sexes. This calls for specific tailored interventions aimed at younger men with less education, Arab men, and young immigrants. PMID- 15542875 TI - Sociodemographic and health behaviour factors associated with obesity in adult populations in Estonia, Finland and Lithuania. AB - BACKGROUND: The aim of this study was to analyse the pattern of sociodemographic variations in the prevalence of obesity in Estonia, Finland and Lithuania. In addition, the association between obesity and selected health behaviours was examined. METHODS: Cross-sectional surveys were conducted among representative national samples of adult populations in 1994, 1996 and 1998. The number of participants aged 20-64 was 3759 in Estonia, 9488 in Finland and 5635 in Lithuania. The data were obtained from mailed questionnaires (covering sociodemographic characteristics, health behaviour indicators, and self-reported height and weight). Obesity was defined as BMI > or = 30 kg/m2. RESULTS: The prevalence of obesity among men and women was 10% and 15% in Estonia, 11% and 10% in Finland, and 10% and 18% in Lithuania respectively. The prevalence of obesity increased only among Estonian men. Obesity was more prevalent among those aged over 50 in all three countries. It was also more prevalent among the less educated women in all countries and among the less educated men in Finland. Obesity was less prevalent among daily smokers among Estonian men and women and Lithuanian men. Physically inactive Estonian women and Finnish men and women were more likely to be obese. CONCLUSIONS: A significant association was found between low educational level and obesity in women in all the countries, but this association was found for men only in Finland. In Finland obesity was also more consistently associated with indicators of unhealthy behaviour than in Estonia and Lithuania. Thus, even though the social gradient of obesity was broadly similar in all the countries studied, differences emerged with regard to the behavioural correlates of obesity. PMID- 15542876 TI - Overweight and obesity in Greek warship personnel: prevalence and correlations. AB - OBJECTIVE: To estimate the prevalence of obesity among the personnel of a Greek warship and to highlight the potential relationship of the body-mass index (BMI). METHODS: Data on behavioural and demographic characteristics, among 274 men of a Greek warship, aged between 19 and 38 years, were analysed. RESULTS: 26.5% of participants were overweight and 4.7% obese. BMI was correlated with age, and cigarette pack-years, and inverse correlated with the aerobic exercise. No significant associations were found between BMI and alcohol consumption. CONCLUSION: A relatively high proportion of obesity was found in a Greek military population. Intervention and prevention measures should be targeted at the promotion of physical activity and the reduction of the smoking behaviour of personnel. PMID- 15542877 TI - Predictors of disability pension in long-term sickness absence: results from a population-based and prospective study in Norway 1994-1999. AB - BACKGROUND: While several socio-demographic predictors of disability pension (DP) have been identified, less is known about the importance of the medical aspects. METHODS: A representative sample of Norwegian long-term sickness absentees, 2043 women and 1585 men, with detailed diagnostic information based on the International Classification of Primary Care (ICPC) was followed up for 5 years. The date of granting DP was obtained from the Norwegian DP-register and used as the dependent variable in Cox multivariate regression analysis. Medical and socio demographic factors were entered as explanatory variables. RESULTS: Kaplan-Meier estimates of the 5 year risk of DP were 22.9% for the full sample, 22.5% for men and 23.3% for the women. Men on sick leave for mental health disorders had an increased disability risk. Except for pregnancy-related cases, which carried a very low risk for future DP, there was no significant difference between the main diagnostic groups among women. Previous sickness absence increased the disability risk but was significant only for total absence above 20 weeks in the 4 years preceding inclusion. Age was the strongest predictor of future DP. Increasing income decreased the risk, bur not linearly. Unemployment status in the year preceding inclusion increased disability risk for women, but not for men. Among cases with musculoskeletal disorders (54.5% of the sample), subgroups with different disability risks were identified in Cox' regression analysis, with a gender-specific pattern. CONCLUSION: In addition to previously known socio demographic predictors, medical variables were important in identifying sickness absentees with an increased risk of DP. PMID- 15542878 TI - The importance of social characteristics of communities for the medically based disability pension. AB - BACKGROUND: The aim of this study was to look for any possible contextual effect of deprivation at municipality level on the risk of being granted the medically based disability pension, controlled for compositional effects due to spatial concentration of people with a high risk of disability. METHODS: The material consists of the residentiary part of a total Norwegian county population aged 20 54 years without disability pension at baseline, n=40,083. This study was performed as a 10-year follow-up study. The relative risk of being granted a disability pension was estimated by logistic regression analyses as odds ratios (OR) between people living in different municipalities according to a municipality deprivation index at three levels, adjusted for individual factors. RESULTS: The OR of disability pension was 1.36 (1.22, 1.51) for people residing in intermediate deprived municipalities and 1.48 (1.31, 1.67) for people residing in the most deprived municipalities compared to the most affluent municipalities, adjusted for gender and age. After adjustment for individual risk factors the OR was 1.26 (1.12, 1.41) and 1.18 (1.04, 1.35) respectively. Analyses stratified by gender showed that the increased risk of receiving a disability pension for men in the most deprived municipalities was explained by individual factors alone. CONCLUSION: Relative municipality deprivation seems to account for an increase in the incidence of disability pension. This effect contributes to marginalization of people living in less affluent areas out of employment and thus to widening socioeconomic inequalities in the population. PMID- 15542879 TI - A comparison of socio-economic differences in long-term sickness absence in a Japanese cohort and a British cohort of employed men. AB - OBJECTIVES: To compare the magnitude of socio-economic differences in sickness absence rates between a Japanese cohort and a British cohort. To assess the effects of self-rated health and behavioural risk factors on sickness absence in the two cohorts, and whether they explain socio-economic differences in sickness absence within and between cohorts. METHODS: An 8 year follow up study of sickness absence in 2504 Japanese male employees in a factory in Japan and 6290 British male employees in civil service departments in London. The rates of first occurrences of long-term (>7 calendar days) sickness absence were determined and compared between these cohorts. Socio-economic status was measured with hierarchical employment grades. RESULTS: The first time sickness absence rates were about two times higher among British men as compared with Japanese men. The rate ratio of lower to higher employment grade was 1.2, 1.3 and 2.1 among Japanese white-collar, Japanese blue-collar and British white-collar employees respectively. Baseline self-rated health and smoking habit predicted sickness absence in both cohorts. After adjusting for these factors a significant difference between the Japanese and British cohorts, and between employment grades remained. CONCLUSIONS: The rate of long-term sickness absence was higher in the British cohort than the Japanese cohort. PMID- 15542880 TI - Smoking from adolescence to adulthood: the effects of parental and own socioeconomic status. AB - BACKGROUND: The aim of the study was to examine the effects of parental socioeconomic status, own socioeconomic status and social mobility upon the development of smoking from adolescence to adulthood. METHODS: Subjects were the participants of the North Karelia Youth Project study from six schools in Eastern Finland. At the baseline in 1978 they were 13 year-olds (n=903) and in the last of the six surveys in 1993 they were 28-year-olds. The parents were studied in 1978 and 1980. The association between smoking and socioeconomic status was measured by education, occupation and income in adolescence and adulthood, and social mobility was measured by the difference between parental and own education. RESULTS: In general, parental socioeconomic status was not significantly associated with the subjects' smoking in adolescence or adulthood. Own socioeconomic status measured at the age of 21 and 28 was strongly related to smoking. Those who were most educated in adulthood had smoked the least already from the age of 13. Social mobility was not significantly associated with smoking. CONCLUSION: The study stresses the importance of own socioeconomic status in relation to smoking, but parental socioeconomic status or social mobility does not have direct effects on smoking. Socioeconomic differences in smoking should be understood as an important determinant for health inequalities. PMID- 15542881 TI - Contribution of deaths related to alcohol or smoking to the gender difference in life expectancy: Finland in the early 1990s. AB - BACKGROUND: In comparison to most other countries in Western Europe, Finland ranks high with regard to male excess mortality. This study examined the contribution of smoking and alcohol to the gender difference in mortality in Finland during 1991-93 among the population aged 15 or over. METHODS: The study is based on data from linked registers. The number of alcohol-related deaths was assessed on the basis of information included in the death certificate, while the conventional method of population attributable fraction was applied to estimate the aggregated contribution of smoking. In combining the effects of alcohol and smoking, three alternative assumptions of their overlap were used. RESULTS: Approximately 51-56% of the gender difference of 7.6 years in the life expectancy at age 15 was estimated to be attributable to alcohol or smoking, depending on the assumption about the overlap of the contribution of alcohol and smoking. CONCLUSIONS: The study confirms the important role of alcohol and smoking as a mechanism contributing to the difference in mortality between men and women in Finland. PMID- 15542882 TI - Impact of cigarette advertising on smoking behaviour in Spanish adolescents as measured using recognition of billboard advertising. AB - BACKGROUND: Cross-sectional studies provide empirical support for associations between advertising and adolescent smoking. The aim of this study was to investigate the relationship between Spanish adolescent smoking behaviour and prior awareness of cigarette advertisements on billboards, using a prospective design. METHODS: 3,664 Spanish children aged 13 and 14 years filled in self completion questionnaires at baseline, and 6, 12, and 18 months later (cohort study). Slides of three advertisements were projected at baseline. A multivariate logistic regression analysis was carried out to detect possible association between number of identified tobacco advertisements brands at baseline and smoking status along time, controlling ASE Model smoking determinants, smoking prevention interventions, age, gender and socio-economic status. RESULTS: The more advertisements identified at baseline, the greater was the risk of being a smoker (p<0.0001). Final percentages of smokers were 15.8%, 16.3%, 19.3%, and 32.6%, respectively, for zero, one, two and three advertisements recognized. When confounders were controlled, the probability of being a smoker increased with the number of advertisements identified [OR 1.26 (95% CI: 1.09-1.46) after 6 months, OR 1.18 (95% CI: 1.03-1.35) after 12 months and 1.15 (95% CI: 1.02-1.30)] after 18 months. It is possible the association would have been even greater if there had not been a differential loss of smokers from the sample. CONCLUSION: Increased awareness of cigarette advertising was associated with a higher smoking incidence and an increased risk of Spanish children becoming smokers. It is, therefore, imperative that cigarette advertising should be banned as a matter of urgency. PMID- 15542884 TI - The million women study: a critique. AB - The regulatory authority in the UK, the Committee on Safety of Medicines, issued advice to health professionals on the day the results of the Million Women Study (MWS) were published. This course of action was undertaken before review of the study by the international medical and scientific community. We have reviewed the methodology of the MWS. It is our belief that the flaws in the design render the results largely uninterpretable because built in biases have affected risk estimates. PMID- 15542885 TI - Is the WHI relevant to HRT started in the perimenopause? AB - The Women's Health Initiative (WHI) hormone replacement therapy (HRT) estrogen plus progestin (E+P) and estrogen-only arms are part of a large NIH-sponsored randomized controlled trial (RCT). Both arms were terminated prematurely after 5 and 8 yr, respectively. The E+P arm showed non-statistically significant increased incidences of cardiovascular events and breast cancer, whereas the E only arm did not. Both arms showed an increased rate of thromboembolic events and stroke. Both arms showed protection against fractures and with protection against colon cancer only in the E+P arm. These results have been widely generalized as indicating a negative risk/benefit ratio for HRT in menopausal women. The WHI results are at odds with results of large epidemiological studies that showed protection against cardiovascular disease. Although the latter data are, in part, confounded by a "healthy user bias," much of the inconsistency may be explained by the fact that women in the latter studies initiated HRT at the menopausal transition, whereas the WHI trial was conducted in older women (mean age 63.3), who were, on average, approx 12 yr postmenopausal. In addition, older trials included women on either unopposed estrogen therapy (ERT) or cyclic HRT regimens. Whatever other forces may have been at work, observational and experimental evidence supports the conclusion that estrogen's atheropreventive effects predominate early, in the absence of vulnerable plaque to be ruptured or thrombotic episodes propagated by narrowed lumens and intravascular turbulence. On the contrary, age-related adverse effects of HRT may prevail once complex atheromas and luminal narrowing/irregularity are established. It is known that prevalence of subclinical "at-risk" atherosclerotic lesions increases in women during the first 5-10 yr after menopause. Furthermore, animal and clinical evidence supports the use of lower doses of estrogen than were employed in the WHI in older/longer postmenopausal women. PMID- 15542886 TI - Adverse neoplastic and cardiovascular outcomes of HRT: the validity of the evidence. AB - The possibility that HRT may increase the risk of cancer, female cancers in particular, has been a general concern ever since HRT was first advocated in the mid-1950s. For cancer of the uterus that concern was vindicated when it was shown that unopposed estrogens greatly increase the risk. However, because HRT is undoubtedly effective in relieving menopausal symptoms, and because it was thought that HRT may reduce the risk of coronary heart disease, the use of estrogens in combination with progestins soon supplanted unopposed estrogen use (if less so among women who had undergone hysterectomies), in the belief, soon confirmed by epidemiological evidence, that combined therapy probably eliminates any increase in the risk of uterine cancer, or perhaps even reduces it. PMID- 15542887 TI - Postmenopausal hormone therapy and breast cancer: a clinician's message for patients. AB - The Women's Health Initiative agrees with some but not all case-control and cohort studies that current use of postmenopausal estrogen-progestin therapy is associated with a small increase in the risk of breast cancer. It is not known whether this is because of new tumor growth or an effect of hormonal therapy on preexisting tumors. Many studies indicate that women who develop breast cancer while using postmenopausal hormone therapy have a reduced risk of dying from breast cancer; this is consistent with an effect on preexisting tumors so that tumors appear at a less virulent and aggressive stage. PMID- 15542888 TI - The rationale for low-dose hormonal therapy. AB - A trend in hormonal therapy has been to use a lower dose. The justification for this approach based on both efficacy and safety is presented. PMID- 15542889 TI - Hormones and bone health in postmenopausal women. AB - Although it has been known for some time that estrogen deficiency is a major pathogenetic factor for osteoporosis related fractures among postmenopausal women, the capability of estrogen (with or without a progestin) to prevent fractures has often been questioned. The publication of the data from the two hormone clinical trials of the Women's Health Initiative lays that discussion to rest. In both studies what have been considered a standard dose of conjugated estrogen with or without medroxyprogesterone acetate significantly reduced the risk of all fractures, including clinical vertebral fractures and hip fracture, in a population of postmenopausal women, average age 63 yr, not selected for osteoporosis by BMD. These results are particularly impressive given the difficulty of finding a fracture benefit in lower risk populations with other anti-resorptive agents. Surrogate data on lower doses of hormone therapy suggest a fracture benefit would be seen if studies were to be done. The other outcomes in WHI make it important to define appropriate clinical guidelines for use of hormone therapy for prevention of fractures in postmenopausal women. PMID- 15542890 TI - New HRT options for the treatment of menopausal symptoms and the maintenance of quality of life in postmenopausal women. AB - Quality of life (QoL) is an important aspect that reflects the way people feel and function. The assessment of QoL quality of life is an essential tool for estimating the efficacy of any therapy in different pathophysiological conditions. In normal, healthy women, the perception of QoL can be significantly modified during the peri/postmenopausal period. Conventional HRT improves not only symptoms, but also QoL. Selection of patients, as well as the choice of a personalized HRT in terms of doses, types, routes of administration, and combination, are the keys to optimize the benefits and reduce the risks. One type of HRT cannot fit all populations of postmenopausal women. The safety and the benefit/risk ratios reported for the standard higher doses used in the past, as well as in the HERS and WHI trials, cannot vaguely be referred to different preparations, and particularly to newer HRT schedules with lower dosages. The demonstration of efficacy of lower-dose HRT provides important information for the treatment of the postmenopausal syndrome. Lower-dose HRT (as 0.3 mg/d of conjugated estrogens) minimizes the side effects and is likely to improve compliance to the treatment. The choice of lower estrogen doses may at least in part reduce the potential risks of postmenopausal hormone use, while maintaining the benefits of conventional HRT. PMID- 15542891 TI - The metabolic basis for the effects of HRT on coronary heart disease. AB - Hormone replacement therapy has profound effects on the cardiovascular system, through metabolic effects as well as direct effects on arterial function. Observational studies showing coronary heart disease benefit have appeared superficially to be at odds with the outcomes from randomized clinical trials, but the lack of benefit from the latter appears to arise from early harm caused by hormone replacement therapy that outweighs any later benefit. The early harm could arise from starting doses of such therapies that are inappropriately high for the women's age, and could therefore cause transient increases in thrombogenesis and adverse vascular remodeling. Later benefit could result from estrogen action on metabolic risk factors, such as lipids, glucose, and insulin metabolism, as well as direct arterial effects reducing atherogenesis. In contrast, this is avoided in observational studies because the women are 15 yr younger and those same starting doses of hormone replacement therapy are appropriate for their age. It is therefore possible that more appropriate hormone replacement therapy regimens, particular in terms of dose, could be devised to effect coronary benefit in older age groups. This concept needs to be tested as a priority before hormone replacement therapies are rejected as being of no cardiovascular benefit. PMID- 15542892 TI - Hormones and coronary atherosclerosis in women. AB - Heart disease is the major health issue facing women in the United States today. Yet, less than 50% of women are aware cardiovascular disease is a health problem. Atherosclerosis begins in childhood and lipid streaks have been identified in girls ages 15-19 in the abdominal aorta and the right coronary artery. Risk factors for cardiovascular disease in women include smoking, diabetes, hypertension, lipid disorders, and menopause. Observational studies have reported a 30-50% reduction in cardiovascular events when estrogen was administered to younger women for menopausal symptoms, yet randomized trials in older patients have failed to show benefit with hormonal replacement therapy. Recent studies have reported preservation of lipid and vascular vasodilatation with low-dose conjugated equine estrogens (CEE) in women and an absence of inflammatory and clotting changes that were observed in high-dose CEE. Recommendations for reducing cardiovascular risk in postmenopausal women include smoking cessation, regular exercise, and weight control. Should hormone therapy be continued beyond management of menopausal symptoms and treatment for osteoporosis, a statin drug should be added to eliminate future cardiovascular complications. Future research will examine low-dose hormonal therapy, earlier administration after menopause, newer agents, and routes of estrogen administration. PMID- 15542893 TI - Postmenopausal hormone therapy: lessons from observational and randomized studies. AB - The effect of estrogen replacement therapy (ERT) and hormone replacement therapy (HRT) for cardioprotection in postmenopausal women remains controversial. Observational studies conducted in the past two decades have suggested an average risk reduction of 50% for the primary prevention of CAD, these findings, however, have not been confirmed by recent randomized clinical trials (RCTs). The discrepancies in results between observational and randomized studies are related to several differences in patient selection, hormone regimen, and biological effect of hormones in different periods of women's life. In an attempt to justify the use of hormone replacement therapy against the mounting contraindications for any use by several opinion leaders and scientific societies, several authors have criticized the design and the results of the randomized clinical trials as the cause of the unexpected results. The randomized clinical studies were conducted exceptionally well; therefore, methodologic issues are not the problem. The main difference between the observational and randomized studies, which may fully explain the discrepancies between these studies, are the women under study and their reasons for taking hormone therapy. In the observational studies women choose to take ovarian hormones initially for menopausal symptoms and then may have decided to continue for other reasons, while in the randomized studies the absence of menopausal symptoms was a pre-requisite for inclusion in the study. This apparently small difference has important implications because symptomatic women are younger and have clinical symptoms that suggest the lack of estrogen effect on several organs or systems. In conclusion, several biological reasons may have contributed to the divergent findings from observational studies and RCTs. Clearly time elapsed since menopause seems to be an important one for its effect on vascular responsiveness to ovarian hormones and to prothrombotic effects. In the meantime, a role remains for combined estrogen and progestin supplementation in the treatment of menopausal symptoms. PMID- 15542894 TI - Hormone replacement in women with breast cancer: the HABITS study. AB - Ever since Professor William T. Creasman suggested the use of hormone replacement therapy in breast cancer survivors in the early 1980s, interest in this field has been guarded but present. Prescribing HRT to breast cancer survivors was initially thought of as being outrageous. Yet even then with experience in HRT spanning a good three decades, and with the breast cancer epidemic, so confidently predicted, then as it is now never actually materializing, doctors working in the field had started to question the conventional wisdom. The debate on whether to treat breast cancer survivors with HRT has been revisited from time to time as there has been a powerful demand for a solution for such symptomatic women. The HABITS study was thus designed to investigate the use of HRT in breast cancer survivors. PMID- 15542896 TI - Identification and analysis of precursors to invasive pancreatic cancer. AB - Histologically distinct noninvasive precursor lesions have been recognized in the pancreas for close to a century. The recent development of a consistent reproducible nomenclature and classification system for these lesions has been a major advance in the study of these noninvasive precursors. The "pancreatic intraepithelial neoplasia" or PanIN system was developed at a National Cancer Institutes sponsored think tank in Park City, Utah. Numerous studies have now demonstrated that genetic alterations in cancer-associated genes are more common in higher grade PanIN lesions then they are in lower grade PanIN lesions, and that higher grade PanIN lesions have many of the same genetic alterations that are found in invasive ductal adenocarcinomas of the pancreas. Thus, just as there is a progression in the colorectal of adenomas to invasive adenocarcinoma, so too is there a progression in the pancreas of histologically low-grade PanIN, to high grade PanIN to invasive ductal adenocarcinoma. PMID- 15542895 TI - Neoplasia of the female reproductive tract: effects of hormone therapy. AB - This review presents the data on the relationship between estrogen and estrogen plus progestin therapy in postmenopausal women and the occurrence of neoplasia in the endometrium, ovary, and uterine cervix. Estrogen only in women with an intact uterus consistently is shown to increase the incidence of endometrial cancer. Estrogen plus a cyclic or sequential progestin reduces the incidence of endometrial cancer to that found in never users. The duration of the progestin administration appears to be important with less than 10 d of progestin having an increased incidence of cancer after 5 yr of therapy. Continuous estrogen plus progestin does not increase the incidence of endometrial cancer. Estrogen and estrogen plus progestin effects on the occurrence of ovarian cancer are inconsistent. The data suggest a possible increase in ovarian epithelial tumors with >10 years use of estrogen only. There is no evidence of a change in the incidence of uterine cervical neoplasia with either estrogen or progestin. PMID- 15542897 TI - Optimal molecular profiling of tissue and tissue components: defining the best processing and microdissection methods for biomedical applications. AB - Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of pancreatic malignancy and other biological phenomena. This chapter reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genotype. The quality of samples isolated from tissue and used for molecular analysis is often glossed-over or omitted from publications, making interpretation and replication of data difficult or impossible. Fortunately, recently developed techniques allow life scientists to better document and control the quality of samples used for a given assay, creating a foundation for improvement in this area. Tissue processing for molecular studies usually involves some or all of the following steps: tissue collection, gross dissection/identification, fixation, processing/embedding, storage/archiving, sectioning, staining, microdissection/annotation, and pure analyte labeling/identification. High-quality tissue microdissection does not necessarily mean high-quality samples to analyze. The quality of biomaterials obtained for analysis is highly dependent on steps upstream and downstream from tissue microdissection. We provide protocols for each of these steps, and encourage you to improve upon these. It is worth the effort of every laboratory to optimize and document its technique at each stage of the process, and we provide a starting point for those willing to spend the time to optimize. In our view, poor documentation of tissue and cell type of origin and the use of nonoptimized protocols is a source of inefficiency in current life science research. Even incremental improvement in this area will increase productivity significantly. PMID- 15542898 TI - Immunohistochemistry and in situ hybridization in pancreatic neoplasia. AB - There are many types of pancreatic neoplasms. Pathologic examination, which includes both routine (e.g., hematoxylin-and-eosin staining) and ancillary (e.g., immunohistochemistry and in situ hybridization) techniques, is essential in correctly typing a pancreatic neoplasm. This chapter focuses on the use of immunohistochemistry and in situ hybridization in the differentiation of pancreatic neoplasms. The materials and methods of these two techniques are described in detail. PMID- 15542899 TI - Practical methods for tissue microarray construction. AB - The tissue microarray (TMA) of Kononen et al. is an extension of an idea originally developed by Battifora and consists of an array of cylindrical cores of paraffin-embedded tissue that are removed from preexisting "donor" paraffin blocks. The donor block is a standard tissue block that may be from surgical pathology, autopsy, or research material. A morphologically representative area of interest within the donor block is identified under the microscope using a stained section (usually hematoxylin and eosin stained) on a glass slide as a guide. The tissue cores are removed from the donor and inserted into a "recipient" paraffin block usually using a custom patented instrument from Beecher Instruments. Using a precise spacing pattern, tissues are inserted at high density, with up to 1000 tissue cores in a single paraffin block. Sections from this block that are cut with a microtome are placed onto standard slides that can then be used for in situ analysis. Depending on the overall depth of tissue remaining in the donor blocks, tissue arrays can generate between 100 and 500 sections. Once constructed tissue microarrays can be used with a wide range of techniques including histochemical staining, immunohistochemical/immunofluorescent staining, or in situ hybridization for either DNA or mRNA. In this chapter we present methods of TMA construction with emphasis on providing detailed tips and techniques. PMID- 15542900 TI - Xenografting and harvesting human ductal pancreatic adenocarcinomas for DNA analysis. AB - Xenotransplantation (xenografting) of primary cancers or cancer cell lines into immunodeficient mice is a commonly used technique to assess tumor growth in response to a variety of experimental agents. When primary pancreatic cancers are xenografted, cancer cells proliferate in the mouse, but the human stroma does not. This growth pattern enables facile genetic analysis of cancer genetics profiles without the contamination of admixed stromal cells typical of primary cancers. PMID- 15542901 TI - Culture and immortalization of pancreatic ductal epithelial cells. AB - Some populations of the epithelial cells from the duct and ductular network of the mammalian pancreas have been isolated and maintained in vitro for up to 3 mo. These cells express many of the surface factors that are unique to them in vivo. They also retain significant drug- and carcinogen-metabolizing capacity in vitro. In this chapter we review the progression of the methods for the isolation, culture and maintenance in vitro for these cells from the earliest when only duct/ductular fragments were obtainable to the current ones which provide epithelial cells. The critical steps in the isolation process are identified and strategies are provided to facilitate these steps. These include the selection of tissue digestive enzymes, the importance of extensive mincing before culture and the importance of roles of some co-factors used in the culture medium. PMID- 15542902 TI - DNA methylation analysis in human cancer. AB - Many tumor suppressor genes (such as p16, Rb, VHL, E-cadherin, and hMLH1) that are silenced by mutation are also inactivated by gene silencing through DNA methylation. Characterization of genes hypermethylated in human cancers but not in normal tissues not only provides insights into cancer biology but also permits the use of methylation-specific polymerase chain reaction-based assays that could serve as diagnostic tests for the early detection and early diagnosis of this disease. To this end, research aimed at the identification and characterization of the methylation status of known and candidate tumor suppressor genes is one strategy for finding putative diagnostic markers. This chapter describes several methods of methylation analysis. PMID- 15542903 TI - Digital single-nucleotide polymorphism analysis for allelic imbalance. AB - Digital single-nucleotide polymorphism (SNP) analysis is developed to amplify a single template from a pool of DNA samples, thereby generating the amplicons that are homogeneous in sequence. Different fluorophores are then applied as probes to detect and discriminate different alleles (paternal vs maternal alleles or wild type vs mutant), which can be readily counted. In this way, digital SNP analysis transforms the exponential and analog signals from conventional polymerase chain reaction (PCR) to linear and digital ones. Digital SNP analysis has the following advantages. First, statistical analysis of the PCR products becomes available as the alleles can be directly counted. Second, this technology is designed to generate PCR products of the same size; therefore, DNA degradation would not be a problem as it commonly occurs when microsatellite markers are used to assess allelic status in clinical samples. Last, digital SNP analysis is designed to amplify a relatively small amount of DNA samples, which is available in some clinical samples. Digital SNP analysis has been applied in quantification of mutant alleles and detection of allelic imbalance in clinical specimens and it represents another example of the power of PCR and provides unprecedented opportunities for molecular genetic analysis. PMID- 15542904 TI - Representational difference analysis as a tool in the search for new tumor suppressor genes. AB - The recognition of a homozygous deletion of genetic material in a tumor genome has been instrumental in several tumor suppressor gene searches. The representational difference analysis (RDA) allows one to identify homozygous deletions even from among the high background of allelic losses that is typical for most cancers. RDA is a polymerase chain reaction (PCR)-based subtractive hybridization method. Two major obstacles to successful enrichment of target sequences from complex genomes were circumvented by RDA. Incomplete reassociation of complex DNA populations is overcome by using representative subpopulations of the tester and driver genomes. In addition, reiterated hybridization, selection, and amplification of the difference products introduces a kinetic component in the enrichment of target sequences. RDA thus enables the identification of homozygous deletions as small as 100 kilobases. Here, we provide a detailed protocol of the RDA procedure, including reflections on frequently encountered technical problems and on the particulars of its application in cancer. PMID- 15542905 TI - Serial analyses of gene expression (SAGE). AB - Serial analysis of gene expression (SAGE) is a molecular biology technique that was developed to measure the global gene expression levels. It has been applied successfully to characterize transcriptomes, compare the transcript levels between normal and diseased tissues, and uncover novel molecules within defined signal transduction pathways. A detailed description is presented in this chapter of the procedures involved to prepare the SAGE libraries. Protocols for automated sequencing and other standard molecular biology techniques can be found elsewhere, and thus are not included herein. PMID- 15542906 TI - Oligonucleotide-directed microarray gene profiling of pancreatic adenocarcinoma. AB - Successful gene profiling studies involve careful experimental design, use of sensitive and accurate technologies, and statistically valid analysis of experimental results. In this chapter we describe our approach to the profiling of pancreatic adenocarcinoma to illustrate the various steps and methods involved in this type of study. Pancreatic adenocarcinoma is a particularly challenging subject for gene profiling, as these tumors have a profound desmoplastic response such that neoplastic epithelium makes up only a small proportion of the tissue mass. We have utilized statistical comparisons of gene expression between adenocarcinoma, normal pancreas, samples of chronic pancreatitis, and pancreatic cancer cell lines that provides a means to deduct the influence of the stromal elements. We utilized oligonucleotide-directed gene chips (Affymetrix), as they allow the simultaneous interrogation of thousands of genes in an efficient, reproducible, sensitive, and highly quantitative manner. The details of the approach we utilized are reported here, including information on experimental design, sample collection, expression level measurements, and data analysis for gene profiling. PMID- 15542907 TI - Identification of differentially expressed proteins in pancreatic cancer using a global proteomic approach. AB - Proteomics is the term used for the large-scale analysis of proteins in biological fluids or cells by biochemical methods. Two approaches are used for proteomics analysis: two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and a mass-spectrometry-based approach, such as surface-enhanced laser desorption ionization (SELDI). SELDI can be used for large protein profiling or peptide identification after enzymatic digestion. In pancreatic cancer, proteomics analysis can be performed with the aim to identify all differentially expressed proteins in cancer cells vs normal pancreatic cells. Protein profiling of pancreatic juice or serum may also identify biomarkers for pancreatic cancer that could be used as diagnostic markers or therapeutic targets. This chapter outlines the use of 2D-PAGE and SELDI for profiling the protein content of pancreatic juice samples and for identifying proteins differentially expressed in pancreatic cancer patient samples compared to control patient samples. PMID- 15542908 TI - Detection of telomerase activity in patients with pancreatic cancer. AB - Telomerase, which ensures the unlimited proliferation by adding TTAGGG repeat at the end of the chromosome, is strongly activated at a very high incidence in a variety of malignant neoplasms including pancreatic cancer. In addition to the acquisition of the immortality, telomerase plays an important role in the aggressive behavior of pancreatic cancer. Invasiveness of human pancreatic cancer cells correlates well with telomerase activity. Exposure of pancreatic cancer to anticancer drugs up-regulates telomerase activity, and the increase in telomerase activity correlates with resistance to the drug-induced apoptosis. More important, diagnositic values of telomerase activity are highly focused because of the lack of other specific genetic markers for pancreatic cancer. Samples of pancreatic juice are obtained at endoscopic retrograde pancreatography using a balloon catheter after intraveneous injection of secretin. Because the pancreatic juice has strong protease and RNase activity, addition of protease inhibitors and RNase inhibitors in the telomerase extraction buffer is necessary for the detection of telomerase activity in the pancreatic juice. A telomeric ladder was detected in 80% patients with carcinoma, whereas only 4.3% patients with adenoma and none with chronic pancreatitis showed positive telomerase activity. PMID- 15542909 TI - Serological analysis of expression cDNA libraries (SEREX): an immunoscreening technique for identifying immunogenic tumor antigens. AB - SEREX (serological analysis of recombinant tumor cDNA expression libraries) is a technique designed to isolate tumor antigens that have elicited high-titer IgG responses in human hosts. This is an immunoscreening method for gene cloning, with two key features that distinguish it from earlier immunoscreenings used to identify targets in autoimmune diseases. First, the assay was designed, at last originally, to analyze autologous immunological responses to cancer, that is, the tumor cDNA library and the screening serum were obtained from the same patient. Second, the screening is performed with serum samples at high dilution (1:1000 1:100), and the secondary antibody used was specific for human IgG. This ensures that only antigens eliciting high-titer IgG responses will be isolated. This latter feature is important in that a key purpose of SEREX is to identify tumor antigens that have elicited both humoral and cell-mediated immune responses in cancer patients. Several tumor antigens identified by SEREX on various types of cancer indeed showed such characteristics and these antigens are being tested as targets for therapeutic cancer vaccines. PMID- 15542910 TI - Modeling pancreatic cancer in animals to address specific hypotheses. AB - Multiple experimental approaches have been employed to study exocrine pancreatic cancer, including the use of animals as surrogates for the human disease. Animals have the advantage that they can be manipulated to address specific hypotheses regarding mechanisms underlying this disease. Implicit in this opportunity is the necessity to match the question being asked with an appropriate animal model. Several approaches to modeling pancreatic cancer have been established that involve animals. First, xenogeneic cell transplantation, generally into immunocompromised rodent subcutis or pancreas, allows examination of (1) the effect of host environment on human or rodent pancreatic cancer cells, (2) whether specific genetic changes in donor cells correlate with certain cancer cell behaviors, and (3) novel approaches to cancer therapy or imaging of tumor growth. Second, carcinogen administration, typically to hamster or rat, allows examination of whether specific genetic, biochemical, cellular, and tissue phenotypic changes, including progression to neoplasia, accompany exposure to a particular chemical. Third, genetically engineered animals, usually transgenic or gene targeted mice, allow examination of (1) whether genetic changes, including oncogene overexpression/mutation or tumor suppressor gene loss, can increase the risk for neoplastic progression, (2) whether specific genetic changes can cooperate during pancreatic carcinogenesis, and (3) how the genetic signature of a neoplasm correlates with particular biological aspects of tumor initiation and progression. Collectively, these experimental approaches permit detailed exploration of pancreatic cancer genetics and biology in the whole animal context, thereby mimicking the environment in which human disease occurs. PMID- 15542911 TI - Strategies for the use of site-specific recombinases in genome engineering. AB - Conventional gene targeting has been very useful in the study of gene function and regulation in mice. However, the methodologies involved have several limitations. First, mutations that cause embryonic lethality largely preclude studies of gene function at a later stage in development. Second, conditional and/or tissue-specific alterations of gene expression cannot be achieved using these methods. In addition, classical gene targeting can be difficult and time consuming. Strategies that make use of site-specific recombinases such as Cre and/or Flp have been developed in recent years to overcome these limitations. These new techniques include global and conditional knockouts, recombinase mediated DNA insertion (RMDI), and recombinase-mediated cassette exchange (RMCE). Together, they have tremendously increased the number and variety of genetic manipulations that can be achieved. PMID- 15542912 TI - Primary explant cultures of adult and embryonic pancreas. AB - The developmental plasticity of adult pancreas is evidenced by the ability to undergo conversion between different epithelial cell types. Specific examples of such conversions include acinar to ductal metaplasia, ductal to islet metaplasia, and generation of ductal structures within islets. Although 90% of human pancreatic cancers are classified as ductal adenocarcinoma, markers of all pancreatic epithelial cell types (acini, ductal, and endocrine) as well as markers of gastric and intestinal lineages can be detected in these tumors. In recent years considerable knowledge has been gained regarding regulation of cellular differentiation and various signaling pathways involved in normal and neoplastic pancreas through studies of pancreatic cancer and immortalized ductal cell lines. However, these studies provide little insight into the context of normal developmental cues, the disruption of which leads to organ pathology. Here we have described a detailed method for preparation, maintenance, and manipulation of adult and embryonic mouse pancreas. These methods may be utilized in studies involving normal epithelial differentiation, contributing to improved understanding of pancreatic development and disease. PMID- 15542913 TI - Zebrafish as a model for pancreatic cancer research. AB - Elucidation of basic mechanisms that regulate pancreatic organogenesis may help define molecular pathways involved in the development of exocrine pancreas cancer. The zebrafish has emerged as a powerful model for genetic dissection of the mechanisms underlying vertebrate organogenesis including formation of the pancreas. Unique properties of zebrafish enable genetic and embryological analyses not feasible using other vertebrate model organisms. The optical clarity of the zebrafish embryos allows visual detection of markers for pancreatic morphogenesis and cytodifferentiation by whole mount immunohistochemistry and RNA in situ hybridization. This feature, coupled with the accessibility of the externally fertilized zebrafish embryo and the small size and fecundity of adult zebrafish, facilitates large-scale forward genetic screens using chemical or insertional mutagenesis techniques. Furthermore, these properties allow high throughput studies that target functions of known genes via antisense or enforced expression studies. Together, such studies are predicted to identify novel genes, or known genes essential for pancreas development. Work in zebrafish is predicted to complement research performed using other vertebrate model organisms, and may help identify markers that define early stages of pancreatic tumorigenesis as well as potential targets for therapy. PMID- 15542914 TI - Development of a cytokine-modified allogeneic whole cell pancreatic cancer vaccine. AB - The management of patients with pancreatic cancer is a multidisciplinary approach that presents enormous challenges to the clinician. Overall 5-yr survival for all patients remains < 3%. Symptoms of early pancreas cancer are nonspecific. As such, only a fraction of patients are candidates for surgery. While surgical resection provides the only curative option, most patients will develop tumor recurrence and die of their disease. To date, the clinical benefits of chemotherapy and radiation therapy have been important but have led to modest improvements. Tumor vaccines have the potential to specifically target the needle of pancreas cancer cells amidst the haystack of normal tissue. The discovery of pancreas tumor-specific antigens and the subsequent ability to harness this technology has become an area of intense interest for tumor immunologists and clinicians alike. Without knowledge of specific antigen targets, the whole tumor cell represents the best source of immunizing antigens. This chapter will focus on the development of whole tumor cell vaccine strategies for pancreas cancer. PMID- 15542915 TI - Overview of linkage analysis: application to pancreatic cancer. AB - Linkage analysis has aided in the identification of genes involved in many diseases, including several cancers. It relies on using family-based data to detect genetic loci that may harbor disease predisposing genes. Although linkage studies were first designed to find the genes responsible for simple Mendelian diseases (diseases caused by alterations in a single gene), today it is more common for investigators to use linkage analysis to locate genes involved in complex diseases (diseases caused by the independent and joint effects of multiple genes often in conjunction with environmental factors), such as pancreatic cancer. During the past decade linkage analysis has been key step in the identification of several cancer genes, including BRCA2 and STK11, which additional studies have shown also carry an increased risk of pancreatic cancer. However, these known genes explain very little of the observed familial aggregation of pancreatic cancer. While the foundations of linkage analysis are relatively straightforward, the actual implementation of linkage studies, especially for complex diseases such as pancreatic cancer, can be quite difficult. This chapter focuses on the basics of linkage analysis for qualitative traits (affected/unaffected) as could be applied to the study of pancreatic cancer. PMID- 15542916 TI - Drop-in, drop-out allele-specific PCR: a highly sensitive, single-tube method. AB - Allelotyping large numbers of samples by allele-specific polymerase chain reaction (PCR) can be problematic if the DNA samples to be tested are of highly variable concentration. On the one hand, analysis of dilute DNA samples often requires nested PCR to produce a product of sufficient yield to be detectable on ethidium bromide-stained agarose gels. Such two-step assays require additional reagents, are labor-intensive, and have a higher risk of contamination. On the other hand, the specificity of allele-specific PCR assays can be lost at high input DNA concentrations. Large population-based genetic studies using DNA from varied sources would benefit from one-tube assays that could detect mutations in samples over a wide range of concentration. We describe a one-tube nested allele specific PCR-based assay, in which the input DNA concentration has little effect on the assay's yield or specificity. An assay using this method is highly sensitive and specific, and was used to type several thousand DNA samples, obtained from various sources, for a G to A transition at human transthyretin codon 122. Similar assays could be readily adapted to any high-throughput allelotype assay where input DNA is of highly variable concentration. PMID- 15542917 TI - Synthesis and assembly of anthrax lethal factor-cholera toxin B-subunit fusion protein in transgenic potato. AB - A DNA encoding the 27-kDa domain I of anthrax lethal factor protein (LF), was linked to the carboxyl terminus of the cholera toxin B-subunit (CTB-LF). The CTB LF fusion gene was transferred into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated in vivo transformation methods and antibiotic-resistant plants were regenerated. The CTB-LF fusion gene was detected in transformed potato leaf genomic DNA by polymerase chain reaction (PCR)-mediated DNA amplification. Immunoblot analysis with anti-CTB and anti-LF primary antibodies verified the synthesis and assembly of biologically active CTB-LF fusion protein oligomers in transformed plant tuber tissues. Furthermore, the binding of CTB-LF fusion protein pentamers to intestinal epithelial cell membrane receptors measured by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA) indicated that the CTB-LF fusion protein made up approx 0.002% of the total soluble tuber protein. Synthesis of CTB-LF monomers and their assembly into biologically active CTB-LF fusion protein pentamers in potato tuber tissues demonstrates the feasibility of using edible plants for production and delivery of adjuvanted LF protein for CTB-mediated immunostimulation of mucosal immune responses against anthrax toxin. PMID- 15542918 TI - Expression of an antimicrobial peptide identified in the male reproductive system of rats. AB - Bin1b is a beta-defensins-like molecule originally isolated from the rat epididymis. Owing to its bactericidal activity, Bin1b may have therapeutic properties suitable for the treatment of sexually transmitted diseases. The amino terminus of the mature Bin1b peptide contains a conserved myristoylated Gly residue. We studied the requirement of the terminal myristoylated Gly residue in the bactericidal activity of Bin1b and found that the terminal myristoylated Gly residue is not essential for the bactericidal activity. In addition, we expressed the tandem repeats of Bin1b in Escherichia coli and found that two tandem repeats of Bin1b protein were successfully expressed. The bacterially expressed tandem Bin1b repeats may be used in a diverse array of biochemical and cell biological studies. PMID- 15542919 TI - A modified hybridization protocol for low-density diagnostic DNA microarrays. AB - To promote the application of DNA microarrays for clinical diagnosis, the problems of cross-hybridization and low signal intensity in the hybridization processes has been addressed. We tested a new hybridization protocol for low density diagnostic DNA microarrays, by skipping the purification step during sample labeling, while elevating the hybridization temperature from 42 degrees C to 52 degrees C, adding a step of distilled water rinsing immediately after hybridization and before the low stringency washing steps. It was found that the modified hybridization protocol works well in our study, which increased detection sensitivity and eliminated nonspecific signals. PMID- 15542920 TI - Knockdown of mouse adult beta-globin gene expression in MEL cells by retrovirus vector-mediated RNA interference. AB - RNA interference (RNAi) efficiently induces sequence-specific gene silencing in mammalian cells through short interfering RNA (siRNA) of 21-23 nucleotides synthesized in vitro or expressed by DNA-based vector. However, introduction of siRNA into mammalian cells by transfection limits the application of RNAi, especially when it is necessary to generate long-term gene silencing in vivo. Virus vector-mediated RNAi provides an alternative to transfection. In the present study, we investigated such transduction system and showed that retrovirus vector-mediated RNAi can substantially down-regulate expression of mouse adult beta-globin gene in MEL cells. The results suggest that retrovirus vector-delivered RNAi may find its use in functional genomics and in gene therapy. PMID- 15542921 TI - An efficient bipartite PCR technique to introduce specific changes in large plasmids. AB - Amplifying an entire double-stranded plasmid by an inverse polymerase chain reaction (PCR) using a pair of tail-to-tail primers is a particularly efficient approach for introducing changes into DNA sequences. However, the approach generally works best for plasmids less than 5 Kb and it can be difficult to amplify the large multicomponent vectors that are used for protein expression in various eukaryotic cells. We have therefore adopted an alternative approach in which two smaller PCR products are generated and then ligated to produce the complete plasmid. A mutagenic primer is used to introduce the desired change and each reaction includes one of a pair of tail-to-tail primers from within an antibiotic resistance gene contained on the plasmid so that the two PCR products contain complementing parts of the complete gene. Ligating the two products generates various combinations but only the correctly ligated molecules recreate the antibiotic resistance gene and are able to replicate in Escherichia coli. When combined with methods to minimize the carryover of template plasmid, this can be an efficient way of introducing mutations into large plasmids. PMID- 15542922 TI - Joint oligogenic segregation and linkage analysis using bayesian Markov chain Monte Carlo methods. AB - One of the most challenging areas in human genetics is the dissection of quantitative traits. In this context, the efficient use of available data is important, including, when possible, use of large pedigrees and many markers for gene mapping. In addition, methods that jointly perform linkage analysis and estimation of the trait model are appealing because they combine the advantages of a model-based analysis with the advantages of methods that do not require prespecification of model parameters for linkage analysis. Here we review a Markov chain Monte Carlo approach for such joint linkage and segregation analysis, which allows analysis of oligogenic traits in the context of multipoint linkage analysis of large pedigrees. We provide an outline for practitioners of the salient features of the method, interpretation of the results, effect of violation of assumptions, and an example analysis of a two-locus trait to illustrate the method. PMID- 15542923 TI - Application of microarrays in high-throughput enzymatic profiling. AB - This review focuses on recent developments in microarray-based technologies for high-throughput screenings of enzymes. Novel methods of protein immobilization, detection of enzymatic activities, and inhibitions were highlighted. PMID- 15542925 TI - Community genetic services in Latin America and regional network of medical genetics. Recommendations of a World Health Organization consultation. AB - The World Health Organization sponsored a Consultation on Community Genetic Services and a Regional Network of Medical Genetics in Latin America in Porto Alegre, Brazil, on June 19, 2003. The main recommendations of the meeting included: (a) the call for government funding of services, research and education in medical genetics; (b) the conduct of epidemiological research on the prevalence and types of birth defects, genetic disorders and genetic predispositions to common diseases; (c) the education of health professionals in genetics; (d) the education of genetic professionals in community health and public health genetics; (e) the fostering of interactions between clinical geneticists, public health personnel, primary health care workers and community organizations, and (f) a better planning of regionalized services to avoid duplication and inefficiency. PMID- 15542924 TI - Appropriate glycosylation of recombinant proteins for human use: implications of choice of expression system. AB - One of the commonest and least well understood posttranslational modifications of proteins is their glycosylation. Human glycoproteins are glycosylated with a bewilderingly heterogeneous array of complex N- and O-linked glycans, which are the product of the coordinated activity of enzymes resident in the endoplasmic reticulum and Golgi apparatus of the cell. Glycosylation of proteins is highly regulated and changes during differentiation, development, under different physiological--and cell culture--conditions and in disease. The glycosylation of recombinant proteins, especially those destined for potential administration to human subjects, is of critical importance. Glycosylation profoundly affects biological activity, function, clearance from circulation, and crucially, antigenicity. The cells of nonhuman species do not glycosylate their proteins in the same way as human cells do. In many cases, the differences are profound. Overall, the species most distant to humans in evolutionary terms, such as bacteria, yeasts, fungi, insects and plants--the species used most commonly in expression systems--have glycosylation repertoires least like our own. This review gives a brief overview of human N- and O-linked protein glycosylation, summarizes what is known of the glycosylation potential of the cells of nonhuman species, and presents the implications for the biotechnology industry. PMID- 15542926 TI - Fish protein stimulated the fibrinolysis in rats. AB - OBJECTIVE: We hypothesized that fish protein affects blood coagulation and/or fibrinolysis, and compared the activity and amounts of factors involved in blood coagulation and fibrinolysis in rats fed the fish protein, which was treated to remove water-soluble and ethanol-soluble elements, from sardine (sardine protein). METHODS: In the first experiment, rats were fed for 21 days an AIN-93G based control diet, and diets in which the casein of the control diet was exchanged for sardine protein at 5, 10 and 20% levels. In the second experiment, rats were fed an AIN-93G control diet and diets containing 5% fish oil, 10% sardine protein or both (5% fish oil + 10% sardine protein) for 21 days. At the end of the experiments, blood coagulation time, hemostatic parameters and fibrinolysis parameters were measured. RESULTS: The activated partial thromboplastin time (APTT), which is an assay for blood coagulation time in the intrinsic blood coagulation pathway, of rats fed the 20% sardine protein diet was significantly prolonged compared to that of rats fed the control diet. The prolonged APTT by dietary sardine protein was due to a significant decrease of the activities of plasma blood coagulation factors VIII, IX, XI and XII. On the other hand, dietary sardine protein significantly increased the activity of tissue-type plasminogen activator, and the amount of plasma plasmin-alpha(2) plasmin inhibitor complex, which are markers of activated plasmin. Moreover, we observed that the 20% sardine protein diet increased the amount of plasma D dimer, which is a degraded product of the fibrin polymer by plasmin. In the second experiment, the APTT and PT of rats fed the F diet were prolonged compared to those of rats fed the control diet, however the concentration and amount of fibrinolytic parameters in the plasma were almost the same as those of rats fed the control diet. In contrast, the F+S diet not only prolonged APTT and PT, but also increased the concentration and amount of fibrinolytic parameters in plasma. CONCLUSIONS: We consider that the beneficial effects to health and amelioration of cardiovascular and cerebrovascular diseases by fish consumption are caused by a combination of the suppressing effect on blood coagulation of n-3 polyunsaturated fatty acids and the promoting effect on fibrinolysis of fish protein. PMID- 15542927 TI - Excretion of vitamin A in urine of women during normal pregnancy and pregnancy complications. AB - BACKGROUND/AIMS: The renal function, including the excretion of low-molecular weight proteins, changes during pregnancy and may cause a urinary excretion of retinol-binding protein (RBP). Whether it is accompanied by a substantial loss of vitamin A (retinol) has not been established yet. We therefore determined the excretion of retinol and RBP in urine of pregnant women. METHODS: The study involved analyses of urine samples from 40 healthy pregnant women and 29 women with pregnancy complications during the third trimester. Analyses of plasma and urine of 7 healthy women and 5 women with pregnancy complications were also carried out 6 weeks antepartum, at time of delivery and 1 week postpartum. RESULTS: Urinary retinol was higher in women who suffered from pregnancy disorders with an influence on maternal metabolism (p < 0.01). RBP was excreted at substantial concentrations in the urine of all 69 women, but there were no differences between the groups. Women with a concomitant excretion of retinol had higher levels of urinary RBP than those without a retinol excretion (p < 0.05). Differences in plasma retinol and RBP were not significant. CONCLUSION: The excretion of urinary retinol may increase significantly during pregnancy complications, which needs further clarification to which extent this condition may negatively affect the vitamin A status in such women. PMID- 15542928 TI - Characterization of breast milk received by infants with gross blood in stools. AB - OBJECTIVE: The aim of this study was to ascertain factors that might be protective of the appearance of gross blood in the stools of breast-fed infants. METHODS: Logistic regression models were formed to search for variables possibly explaining the condition. In addition to the analyzed breast milk factors, mother's allergic disease was introduced into the models to control for its possible confounding effect. The breast milk samples, collected from mothers of infants with gross blood in stools (n = 23) and from mothers of healthy age matched infants (n = 71), were analyzed for concentrations of transforming growth factor-beta2, tumor necrosis factor-alpha, interleukin (IL)-4, IL-10, prostaglandin (PG)E2, cysteinyl leukotrienes (Cys-LTs) and fatty acid composition. RESULTS AND CONCLUSIONS: Increase in the concentrations of PGE2 and Cys-LTs in the breast milk together with mother's allergic disease reduced the likelihood of gross blood in stools in the breast-fed infant. The results suggest that no single factor, but a combination of immunomodulatory factors may protect the child from gross blood in the stools of breast-fed infants. Allergic disease was not a risk factor as mother's allergic disease appeared to counterbalance the gross blood in stools. Due to the preliminary nature of the study, the results need to be verified in a larger setting. The challenge for the future lies in identifying of such active compounds for dietary modification to enforce particularly the properties of the breast milk which are immunoprotective for the infant and to reduce the likelihood of intestinal disorders in at risk infants. PMID- 15542929 TI - Serum C-reactive protein (CRP) levels and insulin resistance in non-obese women with polycystic ovarian syndrome, and effect of bicalutamide on hirsutism, CRP levels and insulin resistance. AB - BACKGROUND/AIMS: Insulin resistance is associated with serum C-reactive protein (CRP) levels. We aimed to evaluate the effect of bicalutamide on insulin resistance and serum CRP levels in non-obese polycystic ovarian syndrome (PCOS) patients. METHODS: 40 non-obese patients (BMI < or =25 kg/m2) with PCOS and, 40 age- and BMI-matched healthy women were studied. Patients received bicalutamide orally at the dose of 25 mg/day. Serum CRP levels were measured with immunometric assay. Homeostasis model assessment (HOMA-IR) index was used for insulin resistance. RESULTS: Mean Ferriman-Gallwey score (FGS) (p = 0.001), insulin (p = 0.001), serum glucose (p = 0.001), prolactin (p < 0.003), total (p < 0.04) and free testosterone (p = 0.001) and free androgen index (FAI) levels (p = 0.001) of PCOS subjects were higher than in the control group. Mean HOMA-IR of PCOS patients was higher than in control subjects (2.43 +/- 1.2 and 0.94 +/- 0.37, p = 0.001). CRP levels in subjects with PCOS was also higher than in control subjects (4.27 +/- 1.33 and 0.98 +/- 0.19, p = 0.001). After bicalutamide treatment, FGS, free and total testosterone and FAI decreased (p = 0.001). HOMA-IR, prolactin and CRP levels did not show any statistical difference with bicalutamide treatment. CONCLUSIONS: PCOS patients had insulin resistance and a high CRP level. Bicalutamide treatment did not influence insulin resistance and CRP level in PCOS, and this ineffectiveness of bicalutamide on CRP levels may be the result of insulin resistance and/or high prolactin levels at this time. PMID- 15542930 TI - Influence of smoking on serum and milk of mothers, and their infants' serum insulin-like growth factor-I and insulin-like growth factor binding protein-3 levels. AB - OBJECTIVE: To investigate the serum and milk in active-smoking and nonsmoking mothers, and their infants' insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding protein-3 (IGFBP-3) levels. DESIGN AND SETTING: A cohort study conducted at a tertiary medical center. SUBJECTS: Forty-four mothers (age range: 21-34 years) and their newborns (7 days old) were enrolled in the study. Mothers were interviewed and classified according to their smoking status into one of two groups: the active-smoking mothers (n = 21) and the nonsmoking mothers (n = 23). RESULTS: There was no difference noted in either IGF-I, IGFBP-3 or IGF I/IGFBP-3 ratios in serum and milk of mothers, and their infants' serum samples according to maternal smoking. CONCLUSION: This study demonstrated that maternal smoking (5-10 cigarettes/day) did not influence the maternal and infant serum levels of IGF-I and IGFBP-3 as well as the breast milk levels of these peptides. PMID- 15542931 TI - Clinical-biochemical correlations in acromegaly at diagnosis and the real prevalence of biochemically discordant disease. AB - OBJECTIVE: To analyze clinical-biochemical correlations in newly diagnosed acromegaly, focusing in particular on patients with discrepant parameters. DESIGN: Retrospective study. METHODS: Data from 164 patients with acromegaly seen between 1995 and 2003. Patients were reviewed for the presence of headaches, arthralgias, hypertension, menstrual abnormalities, impotence, glucose intolerance or diabetes. Biochemical evaluation consisted of age- and gender adjusted IGF-I levels and glucose-suppressed GH. RESULTS: Magnetic resonance imaging (MRI) revealed macroadenoma in 127 patients and microadenoma in 37. Patients with macroadenomas were younger than those with microadenomas and the disease was more frequent in females. Excluding acral enlargement, which was present in all the patients, the most commonly reported complaints were headaches (66%) and arthralgias (52%). Hypertension was present in 37% of patients, whereas the prevalence of glucose intolerance and diabetes was 27 and 32%, respectively. Hyperprolactinemia was present in 20% of patients with microadenomas and in 40% of patients with macroadenomas. Hypogonadism was demonstrated in more than half of the patients and was not related to tumor size or prolactin level. Of all the clinical and metabolic abnormalities of acromegaly, only the presence of diabetes correlated with both basal and nadir post-glucose GH levels. Only 4 patients (2.4%) had glucose-suppressed GH values of <1 ng/ml in the presence of clinical evidence of acromegaly, an elevated IGF-I level and a pituitary adenoma on MRI. CONCLUSIONS: Clinical features of acromegaly correlate poorly with indices of biochemical activity. The prevalence of biochemically discordant acromegaly is considerably lower than recently reported. PMID- 15542932 TI - Radiosurgical planning of meningiomas: compromises with conformity. AB - The radiosurgical planning of meningiomas frequently necessitates compromises between irradiating tumour and risking damage to adjacent structures. In selected cases, we resolved this by excluding part of the tumour from the prescription isodose volume. Most of these compromises or 'suboptimal' plans achieved growth control. Growth control could not be related to conformity indices or to various measures of the radiation dose received by the meningioma. Examining recurrences, 75% arose from dura outside the original treatment field. These findings are discussed in terms of dose prescription protocols and the use of conformity indices in planning. The importance of the dural origin of meningiomas is well established in surgical practice, as reflected by Simpson's grades, but may be equally significant in radiosurgical practice. PMID- 15542933 TI - Increase in collagen turnover but not in collagen fiber content is associated with flow-induced arterial remodeling. AB - BACKGROUND: Degradation and synthesis of collagen are common features in arterial geometrical remodeling. Previous studies described an association between arterial remodeling and an increase in collagen fiber content after balloon injury. However, this does not exclude that the association between collagen content and remodeling depends on arterial injury since the association of collagen fiber content and arterial remodeling, without arterial injury, has not been investigated. The aim of the present study was to study the relation between flow-induced arterial geometrical remodeling, without arterial injury, and collagen synthesis and degradation, collagen fiber content and cell-migration associated moesin levels. METHODS AND RESULTS: In 23 New Zealand White rabbits an arteriovenous shunt (AV shunt) was created in the carotid and femoral artery to induce a structural diameter increase or a partial ligation (n = 27 rabbits) to induce a diameter decrease. In both models, arterial remodeling was accompanied by increased procollagen synthesis, reflected by increased procollagen mRNA or Hsp47 protein levels. In both models, however, no changes were detected in collagen fiber content. Active MMP-2 and moesin levels were increased after AV shunting. CONCLUSIONS: Collagen synthesis and MMP-2 activation were associated with arterial remodeling. However, a change in collagen fiber content was not observed. These results suggest that, during flow-induced geometrical arterial remodeling, increases in collagen synthesis are used for matrix collagen turnover and cell migration but not to augment collagen fiber content. PMID- 15542934 TI - Effects of combined estriol/pravastatin therapy on intima-media thickness of common carotid artery in hyperlipidemic postmenopausal women. AB - BACKGROUND: Several studies show that 17beta-estradiol (E2) has protective effects on atherosclerosis in the arterial wall in postmenopausal women. Little information is, however, available regarding the effect of estriol (E3) on atherosclerosis. This study was conducted to investigate the effects of E3 alone and combined E3/pravastatin therapy on intima-media thickness (IMT) of common carotid artery in postmenopausal women. METHODS: Thirty-three postmenopausal women were allocated to four groups: daily treatment with E3 (2 mg) alone (E3 group, n = 10), pravastatin (10 mg) alone (pravastatin group, n = 6), combined treatment with E3 (2 mg) and pravastatin (10 mg; E3/pravastatin group, n = 7) and untreated control group (n = 10). All women attended the Kobe University Hospital once a year for routine gynecological and ultrasonographic examinations for the evaluation of atherosclerosis. RESULTS: A significant decrease in IMT was noted in the E3/pravastatin group compared with that in the untreated control group (p < 0.05), whereas there was no significant difference in the reduction rate of IMT in the pravastatin group, E3 group and untreated control group. CONCLUSIONS: The combined E3/pravastatin therapy appeared to retard the progression of atherosclerosis in postmenopausal women. PMID- 15542935 TI - Plasma leptin concentration in tamoxifen-treated ovariectomized rats. AB - Leptin, the ob gene product, has an important role in the regulation of body weight. Although tamoxifen, a nonsteroidal antiestrogenic agent, is known to have estrogenic effects on fat metabolism, its influence on adipose tissue remains unknown. In the present study, the effect of tamoxifen on the concentration of leptin was investigated in ovariectomized rats treated with tamoxifen or vehicle. The dosage of tamoxifen was extrapolated from the human dosage. Food intake, adipose tissue weight, and plasma insulin were assessed at the end of the experiment. RESULTS: Tamoxifen-treated rats showed a significant reduction of body weight gain, food intake, adipose tissue weight and leptin concentration (p < 0.001). The plasma insulin level was significantly higher after tamoxifen treatment (p = 0.01) in tamoxifen-treated rats than in control rats. We concluded that tamoxifen reduces food intake in the acute phase and a reduction of adipose tissue gain may result in reduced levels of plasma leptin in ovariectomized rats. Furthermore, rats treated with tamoxifen may be resistant to insulin action. PMID- 15542936 TI - The major histopathologic characteristics in the vulvar vestibulitis syndrome. AB - OBJECTIVE: In order to better understand the etiology of the vulvar vestibulitis syndrome, we examined the histopathologic parameters in vestibular mucosa, and compared the findings in specimens obtained from women with vulvar vestibulitis with those obtained from the control group. STUDY DESIGN: Specimens of vestibulitis were obtained from 24 patients, undergoing circumferential vestibulectomy under general anesthesia due to the symptoms and signs consistent with vulvar vestibulitis. Control specimens were obtained from 16 women, without symptoms or signs of vulvar vestibulitis, undergoing reconstructive introital surgery due to roomy vagina, rectocele or painful episiotomy. All vestibular tissue specimens were examined for intensity of inflammation, extension of inflammatory cells into the epithelium, vascular proliferation, the presence of mast cells and proliferation of peripheral nerve bundles. RESULTS: No significant difference was found regarding the degree of inflammation, the extension of inflammatory cells into the epithelium, the vascular proliferation and the presence of mast cells while comparing the study and the control groups. The only histopathologic feature, differentiating the patients with vulvar vestibulitis from the control group, was the proliferation of peripheral nerve bundles found in 19 out of 24 (79.1%) specimens expressing vestibulitis and in none of 16 control specimens (p < 0.0001). CONCLUSION: Our results, therefore, support the existence of peripheral nerve hyperplasia in vestibular tissue obtained from patients with vulvar vestibulitis, and exclude the role of active inflammation or mast cells as probable etiologies for the vulvar vestibulitis syndrome. PMID- 15542937 TI - Is the mycobacteria-derived purified protein response in atopic asthmatic children different? AB - BACKGROUND: The response to mycobacteria-derived purified protein (PPD) is mediated primarily by T-helper-1 response and is expected to be inhibited in atopic diseases. The aim of this study was to investigate whether the PPD response is different in atopic asthmatic children. METHODS: 40 atopic asthmatic children (mean age 8.3 +/- 4.9 years) and 40 healthy age- and sex-matched children who had received bacillus Calmette-Guerin (BCG) vaccination were included in the study. Five PPD units were administered intradermally to all children and were interpreted after 48 h. RESULTS: There was no correlation between serum total IgE level and PPD induration (p=0.054). The PPD induration was not statistically different between the children who used inhaled corticosteroid and those who did not. Although the PPD positivity (induration > or =5 mm) rate was higher in atopic asthmatic children (50%) than in healthy children (32.5%), the difference was not found to be statistically significant. The PPD induration in atopic asthmatic children (7.41 +/- 5.58 mm) was found to be greater than the one in healthy children (5.21 +/- 3.39) (p < 0.039). The induration in atopic asthmatic children (5.21 +/- 3.77) and healthy children (4.43 +/- 2.32) did not show a difference in children who where vaccinated only once with BCG, but it was found to be statistically significantly greater in atopic asthmatic children (12.50 +/- 5.90) than healthy children (7.08 +/- 4.70) who were vaccinated with BCG twice (p <0.012). The proportion of having a PPD induration of > or =10 mm was found to be higher in atopic asthmatic children than in the healthy ones (32.5 vs. 12.5%) (p <0.032). CONCLUSION: Our data showed that the PPD response was stronger in BCG-vaccinated atopic asthmatic children than in healthy BCG-vaccinated ones. PMID- 15542938 TI - Mouse models of atopic eczema critically evaluated. AB - Atopic eczema (AE) is a chronic relapsing inflammatory skin disorder with increasing prevalence in Western societies. Even though we have made considerable progress in understanding the cellular and molecular nature of cutaneous inflammation, the precise pathomechanisms of AE still remain elusive. Experimental animal models are indispensable tools to study the pathogenic mechanisms and to test novel therapeutic approaches in vivo. For AE a considerable number of mouse models have been proposed and have been used to study specific aspects of the disease, such as genetics, skin barrier defects, immune deviations, bacteria-host interactions or the role of cytokines or chemokines in the inflammatory process. While some models closely resemble human AE, others appear to reflect only specific aspects of the disease. Here we review the currently available mouse models of AE in light of the novel World Allergy Organization classification of eczematous skin diseases and evaluate them according to their clinical, histopathological and immunological findings. The pathogenetic analogies between mice and men will be discussed. PMID- 15542939 TI - The management of glottic and supraglottic cancers of the larynx in relation to neck metastasis. AB - OBJECTIVES: We evaluated the management of glottic and supraglottic laryngeal cancers in relation to neck metastasis. PATIENTS AND METHODS: Fifty-two patients (51 males, 1 female; mean age 59.2 years; range 27 to 82 years) underwent surgery for primary laryngeal cancers. Preoperative and postoperative TNM classification and staging were made according to the AJCC 1997 criteria. The mean follow-up period was 39.4 months (range 6 to 74 months). RESULTS: The tumors were glottic in 31 patients and supraglottic in 21 patients. Laryngectomies were partial in 34 patients and total in 18 patients. All the patients with supraglottic tumors and those with glottic T2-4 tumors underwent neck dissection, as well. Radiotherapy was administered to 22 patients with established N2 or N3 tumors. Clinical assessment was in agreement with the pathological result in 63.5%; 13.5% and 23.1% of cases were underdiagnosed and overdiagnosed, respectively. The rates of neck metastasis were 0% in T1, 25% in T2, 75% in T3, and 66.7% in T4 supraglottic tumors. The corresponding rates for glottic cancers were 0%, 16.7%, 28.6%, and 60%, respectively. The overall rate of N+ tumors was 28.9%. Four patients (7.7%) developed local, two patients (3.9%) developed regional recurrences. Mortality occurred in eight patients (15.4%) due to following causes: laryngeal recurrence (T4N2, T4N2, T3N1), regional recurrence (T4N2), a second primary malignancy in one patient, and other causes in three patients. Five- and two-year disease specific survival rates (Kaplan-Meier analysis) were 90.7%, and overall survival rates were 73.7% and 87.3%, respectively. CONCLUSION: Neck metastasis and advanced stage of the tumor were the most effective prognostic factors. PMID- 15542940 TI - Detection of Epstein-Barr virus DNA by polymerase chain reaction in surgical specimens of patients with squamous cell carcinoma of the larynx and vocal cord nodules. AB - OBJECTIVES: We investigated the incidence of Epstein-Barr virus (EBV) infection in surgical specimens of squamous cell carcinoma of the larynx and vocal cord nodules. PATIENTS AND METHODS: Formalin-fixed, paraffin-embedded tissue samples randomly selected from 22 patients with squamous cell carcinoma of the larynx (20 males, 2 females; mean age 52 years) and from 17 patients with vocal cord nodules (14 males, 3 females; mean age 38 years) were examined by the polymerase chain reaction. The patients were assessed in terms of localization of the disease, smoking habits, duration of smoking, and the presence or absence of EBV DNA. RESULTS: Twenty-one patients with laryngeal carcinoma and nine patients with vocal cord nodules were smokers. Polymerase chain reaction showed EBV DNA in 11 patients (50%) with laryngeal carcinoma and in seven patients (41.2%) with vocal cord nodules. No significant differences were found with respect to EBV DNA positivity, smoking habits, smoking period, the number of cigarettes consumed daily, localization of disease, and tumor stage (p>0.05). CONCLUSION: Epstein Barr virus does not seem to be directly associated with the pathogenesis of laryngeal squamous cell carcinoma, but its role in the etiology of laryngeal proliferative diseases needs more elucidation. PMID- 15542941 TI - Causes of dysphonia in patients above 60 years of age. AB - OBJECTIVES: We evaluated the causes of hoarseness in patients above 60 years of age. PATIENTS AND METHODS: The study included 98 patients (40 females, 58 males; mean age 64 years; range 60 to 77 years) who presented with hoarseness. All the patients were examined by videolaryngoscopy. Temporary hoarseness secondary to respiratory tract infections was excluded. RESULTS: The most common cause of hoarseness was benign vocal fold lesions (28.6%) followed by malignant lesions (27.6%), vocal fold paralysis (25.5%), functional dysphonia (10.2%), and presbyphonia (8.2%). Laryngopharyngeal reflux was found in 13 patients (13.3%). Fourteen male and 11 female patients had paralysis of the recurrent laryngeal nerve. Malignancies that caused hoarseness without paralyzing the vocal folds were larynx carcinoma (n=18), hypopharynx carcinoma (n=8), and multiple malignancy (n=1). CONCLUSION: Our data show that dysphonia develops due to disease processes associated with aging rather than to physiologic aging alone. Considering adverse influences of dysphonia on the quality of life of elderly population, efforts should be directed to elucidation of the cause and to performing appropriate treatment. PMID- 15542942 TI - A case of Ewing's sarcoma in the mandible and the skull base. AB - A thirteen-year-old boy with Ewing's sarcoma was presented. The treatment was considerably delayed because of the initial diagnostic difficulties due to its insidious presentation and the unwillingness of the child's parents to further investigations and treatment. During a three-year delay, the tumor turned out to be a gross painful mass from a painless lesion. Cranial computed tomography and magnetic resonance imaging with contrast showed a destructive lesion extensively involving the right mandible and the skull base, with invasion to the intracranial space. Facial and intratemporal portions of the mass were removed totally, but the intracranial extension could not be totally excised because of cavernous sinus involvement. Histologic and immunohistochemical findings were consistent with the diagnosis of Ewing's sarcoma. The patient received systemic chemotherapy and radiotherapy following surgery. A month after radiation therapy, radiologic investigations showed a lung mass suggestive of metastasis. He died two days after hospitalization, from an intracranial hemorrhage associated with the intracranial mass. PMID- 15542943 TI - A case of carotid body tumor. AB - A thirty-five-year-old woman had an asymptomatic, slow growing mass that pushed the right tonsilla palatina and was pulsatile in the oropharyngeal region. Magnetic resonance imaging showed a vascular mass at the carotid bifurcation, causing displacement of the external and internal carotid arteries. The diagnosis of a carotid body tumor was confirmed by magnetic resonance angiography. Surgery included a subadventitial dissection at the carotid bifurcation, preserving the hypoglossal and vagal nerves. The patient developed loss of function of the tenth cranial nerve postoperatively. After a year follow-up period, there was no change in the function of the tenth cranial nerve. PMID- 15542944 TI - Thyroid metastasis from renal cell carcinoma eight years after radical nephrectomy: a case report. AB - A forty-five-year-old female patient presented with thyroid nodules eight years after radical nephrectomy for renal cell carcinoma (T2N0M0, grade 2, clear cell subtype) in the right kidney. Ultrasonography of the neck showed multiple hypoechoic macronodules. There was no lymphadenomegaly. During total thyroidectomy following an initial diagnosis of multinodular goiter, frozen section analysis was made, which showed metastasis from renal cell carcinoma. Immunohistochemical studies demonstrated that the tumor cells were negative for thyroglobulin staining. No recurrences or other metastatic disease were seen during a follow-up period of 12 months. Although clinically the thyroid gland is a rare site for tumor metastasis, this possibility should be kept in mind in patients with a previous history of renal cell carcinoma. PMID- 15542945 TI - Kimura's disease in the parotid and submandibular regions: two case reports. AB - Two male patients (aged 13 and 50 years) who presented with a complaint of painless mass in the parotid and submandibular regions, respectively, were found to have eosinophilia and increased blood immunoglobulin E (IgE) levels. Following ultrasonographic examination of the masses, surgical excision was performed. Histopathologic evaluation revealed massive lymphocytic and eosinophilic infiltration. Histopathologic findings and the presence of peripheral eosinophilia and high IgE levels led to a diagnosis of Kimura's disease. Postoperatively, no recurrences were detected within a year follow-up. PMID- 15542946 TI - The plerocercus of Ditrachybothridium macrocephalum Rees, 1959 from two deep-sea elasmobranchs, with a molecular analysis of its position within the order Diphyllidea and a checklist of the hosts of larval diphyllideans. AB - New collections were made of Ditrachybothridium macrocephalum Rees, 1959 from the deep-sea elasmobranchs Apristurus laurussonii (Saemundsson) and cf. Rajella bigelowi (Stehmann) in the northern Atlantic Ocean, allowing studies of the morphology of its plerocercus. The material has also allowed an estimate of its position within the Diphyllidea inferred from molecular data for the first time. The plerocercus is withdrawn, but not invaginated, within a cyst with a thick vacuolated wall. The encysted strobila is distinct, with several putative segments and advanced genital anlagen, and the scolex is fully developed with a distinct apical pad. 28S rDNA (D1-D3 regions) sequences were generated from both encysted and excysted D. macrocephalum , as well as from other diphyllidean and pseudophyllidean (outgroup) taxa for use in phylogenetic analyses. Maximum parsimony and Bayesian inference analyses both showed that the diphyllidean genus Echinobothrium is paraphyletic without inclusion of the only two other diphyllidean genera, Ditrachybothridium Rees, 1959 and Macrobothridium Khalil & Abdul-Salam, 1989. A list of published records of larval diphyllideans, their hosts and localities is included. PMID- 15542947 TI - Nematodes from the squeaker fishes Synodontis nigromaculatus and S. vanderwaali from the Okavango River, Botswana, including three new species. AB - Five nematode species were recorded from the stomach and rectum of the spotted squeaker Synodontis nigromaculatus Boulenger or the finetooth squeaker S. vanderwaali Skelton & White (Mochokidae, Siluriformes) from the Okavango River, Botswana: Falcaustra similis n. sp. (Kathlaniidae), Labeonema africanum n. sp. (Cosmocercidae), Synodontisia okavangoensis n. sp. (Pharyngodonidae), Procamallanus ( Procamallanus ) laeviconchus (Wedl, 1861) (Camallanidae) and Spinitectus sp. (Cystidicolidae) (only a single female). F. similis (type-host Synodontis nigromaculatus ) differs from the similar species F. straeleni Campana Rouget, 1961 mainly by the number and disposition of the male caudal papillae and the structure of the mouth; L. africanum (type-host S. nigromaculatus ) differs from its congeners in having distinctly longer spicules (105-120 microm), a relatively shorter gubernaculum (30-36 microm) and in the number and arrangement of the male caudal papillae; Synodontisia okavangoensis (type-host Synodontis nigromaculatus ) is characterised principally by crescent-shaped cephalic papillae, length (87 microm) and shape of the spicule as well as large eggs (0.156-0.180x69-102 microm). Synodontisia moraveci Anderson & Lim, 1996 is transferred to a newly established genus, Royandersonia n. g., (as its type species) as R. moraveci (Anderson & Lim, 1996) n. comb. Some taxonomic problems concerning Procamallanus ( P .) laeviconchus are discussed. All species were studied by scanning electron microscopy for the first time. PMID- 15542948 TI - Two nematodes, Dentinema trichomycteri n. g., n. sp. (Cosmocercidae) and Procamallanus chimusensis Freitas & Ibanez, 1968 (Camallanidae), from catfishes Trichomycterus spp. (Pisces) in Colombia. AB - Light and scanning electron microscopical examinations of nematode samples collected from the stomachs and intestines of catfishes Trichomycterus spp. (Trichomycteridae, Siluriformes) from three streams in Colombia revealed the presence of two species, Dentinema trichomycteri n. g., n. sp. (Cosmocercidae) (type-host Trichomycterus sp.) and Procamallanus ( Spirocamallanus ) chimusensis Freitas & Ibanez, 1968 (Camallanidae). The new, monotypic genus Dentinema is characterised mainly by the presence of a triangular mouth surrounded by three poorly developed lips, four submedian cephalic papillae, three conical teeth in the well-developed buccal cavity, an elongate oesophageal isthmus which is clearly separated from the corpus, precloacal oblique muscle bands, and by the absence of a ventral sucker. P. chimusensis , recorded from Colombia for the first time, is redescribed from specimens collected from Trichomycterus chapmani (Eigenmann) (a new host record) and Trichomycterus sp.; new observations show that this species belongs to a small group of Procamallanus spp. exhibiting a distinct sexual diversity in the structure of the buccal capsule. P. pexatus Pinto, Fabio, Noronha & Rolas, 1976 is synonymised with P. chimusensis . PMID- 15542949 TI - Description of two new species of Glypthelmins Stafford, 1905 (Digenea: Macroderoididae) in Rana spp. from Mexico, based on morphology and mtDNA and rDNA sequences. AB - Glypthelmins Stafford, 1905 includes 29 putative species commonly found in the intestine and liver of anurans from all over the world but mainly in the Americas. Partial sequences of the cytochrome c oxidase subunit 1 ( cox 1), ribosomal internal transcribed spacer region 2 (ITS2) and the large subunit 28S rDNA gene were obtained and analysed using pairwise distance matrices and parsimony methods in order to characterise the interrelationships between 14 isolates of four nominal species of Glypthelmins recognised on morphological grounds. The highest intra-specific sequence divergence occurred in the cox 1 (18.53%) sequence, followed by that of the ITS2 (5.44%) and 28S (4.63%). Genetic variability was detected between the three isolates originally identified as G. facioi Brenes et al., 1959 from two localities in Mexico and one locality in Costa Rica. Sequence divergence exhibited among these isolates ranged from 10.70 to 11.22%, from 0.48 to 0.97% and from 1.33 to 1.88% for cox 1, ITS2 and 28S, respectively. Phylogenetic analysis combining all three data-sets generated a single most parsimonious tree. The three isolates of G. facioi form a clade, with an isolate collected from frogs in Veracruz State as the sister group to an isolate from Tabasco State + G. facioi from Costa Rica. The information derived from pairwise distance of independent data-sets plus the phylogenetic information indicate that each of the two isolates from Mexico, identified a priori as G. facioi, represent separate species. A re-examination of specimens was carried out and a re-evaluation made of the morphological characters to find reliable differences that had been overlooked. As a consequence, G. brownorumae n. sp. from Tabasco and G. tuxtlasensis n. sp. from Veracruz are described based on molecular and morphological differences. PMID- 15542950 TI - Anthobothrium lesteri n. sp. (Cestoda: Tetraphyllidea) in Carcharhinus melanopterus from Heron Island, Australia, with comments on its site, mode of attachment, reproductive strategy and membership of the genus. AB - Anthobothrium lesteri n. sp. in Carcharhinus melanopterus from Heron Island, Australia is described and figured. A combination of characters, namely the number of testes, a weakly laciniate strobila and mature proglottis which is conspicuously longer than wide distinguishes it from other species that appear to be consistent with van Beneden's original concept of Anthobothrium . The scolex and the peduncle are relatively small and frail when compared with the rest of the strobila and with the detached proglottis. A myzorhynchus is absent and the stalked, undivided, simple bothridia have thickened margins. Two circular areas of muscles, which resemble accessory suckers, occur in the centre of each bothridium. There is a distinct cephalic peduncle with spiniform blade-like microtriches followed by craspedote proglottides which have a posterior four lobed velum. Apolysis is well developed and the oncosphere has a polar filament and a dense covering of spines. Comments are made on the site, the mode of attachment, an aspect of the reproductive strategy of the new species in relation to intestinal eversion in some sharks, and membership of the genus. PMID- 15542951 TI - Revision of Allobenedenia Yamaguti, 1963 (Monogenoidea: Capsalidae) with the description of A. zhangi n. sp. from Epinephelus fasciatus (Teleostei: Serranidae) in the South China Sea. AB - Allobenedenia Yamaguti, 1963 is revised to include capsalid species with five radial septa in the haptor. Megalocotyloides Bychowsky & Nagibina, 1967 and Allosprostonia Lawler & Hargis, 1968 are considered junior synonyms of Allobenedenia. Eight described species comprise the genus: A. convoluta (Yamaguti, 1937) Yamaguti, 1963 (type-species), A. epinepheli (Bychowsky & Nagibina, 1967) n. comb., A. patagonica (Evdokimova, 1969) n. comb., A. pedunculata Raju & Rao, 1980, A. pseudomarginata (Bravo-Hollis, 1958) n. comb., A. sebastodi (Egorova, 1994) n. comb., A. yamagutii (Egorova, 1994) n. comb. and A. zhangi n. sp. A. ishikawae (Goto, 1894) Yamaguti, 1963 is considered a species inquirenda; Megalocotyloides hongkongensis Wu, Lu & Woo, 2002 is placed in synonymy with A. yamagutii; M. grandiloba (Paperna & Kohn, 1964) Egorova & Aleshkina, 1984 is provisionally returned to Megalocotyle as M. grandiloba Paperna & Kohn, 1964; and specimens identified as M. grandiloba from Serranus cabrilla (Serranidae) by Buhrnheim et al. (1973) are considered an undescribed species of Allobenedenia. A. zhangi n. sp. is described from the external surface of Epinephelus fasciatus (Serranidae) of Allobenedenia. A. zhangi n. sp. is described from the external surface of Epinephelus fasciatus (Serranidae) in the South China Sea and is characterised by having an elongtate proximally uncoiled male copulatory organ extending to near the level of the anterior margin of the ovary, a large prostatic reservoir apparently external to the male copulatory canal, and a submedial protuberance on the dorsal surface of the accessory sclerite of the haptor. PMID- 15542952 TI - Gigantolaelaps gilmorei Fonseca, 1939 (Acari: Laelapidae): taxonomic status, lectotype and paralectotype designation and new distributional records. AB - A lectotype series is designated for Gigantolaelaps gilmorei Fonseca, 1939 in the Acari Collection of the Instituto Butantan in Sao Paulo, Brazil. This laelapid mite species has a widespread geographical distribution. We report a close association with Oryzomys russatus (Wagner) in southeastern Brazil and Argentina, the most southerly part of its range. In Paraguay, the primary host is O. nitidus (Thomas). PMID- 15542954 TI - Early goal-directed therapy: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for early goal directed therapy that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: During the first 6 hrs of resuscitation of sepsis-induced hypoperfusion, specific levels of central venous pressure, mean arterial pressure, urine output, central venous (or mixed venous) oxygen saturation should be targeted. When central venous oxygen saturation remains low, despite achieving central venous pressure and mean arterial pressure targets, packed red blood cells or dobutamine should be considered. Increasing cardiac index to achieve an arbitrarily predefined elevated level is not recommended. PMID- 15542955 TI - Fluid resuscitation in severe sepsis and septic shock: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for fluid resuscitation in severe sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Fluid resuscitation of severe sepsis may consist of natural or artificial colloids or crystalloids. Fluid challenge should be administered and repeated based on response (increase in blood pressure and urine output) and tolerance (evidence of intravascular volume overload). PMID- 15542956 TI - Vasopressor and inotropic support in septic shock: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for vasopressor and inotropic support in septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and to improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: An arterial catheter should be placed as soon as possible in patients with septic shock. Vasopressors are indicated to maintain mean arterial pressure of <65 mm Hg, both during and following adequate fluid resuscitation. Norepinephrine or dopamine are the vasopressors of choice in the treatment of septic shock. Norepinephrine may be combined with dobutamine when cardiac output is being measured. Epinephrine, phenylephrine, and vasopressin are not recommended as first-line agents in the treatment of septic shock. Vasopressin may be considered for salvage therapy. Low-dose dopamine is not recommended for the purpose of renal protection. Dobutamine is recommended as the agent of choice to increase cardiac output but should not be used for the purpose of increasing cardiac output above physiologic levels. PMID- 15542957 TI - Diagnosis of infection in sepsis: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for the diagnosis of infection in sepsis that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSIONS: Obtaining a precise bacteriological diagnosis before starting antibiotic therapy is, when possible, of paramount importance for the success of therapeutic strategy during sepsis. Two to three blood cultures should be performed, preferably from a peripheral vein, without interval between samples to avoid delaying therapy. A quantitative approach is preferred in most cases when possible, in particular for catheter-related infections and ventilator-associated pneumonia. Diagnosing community-acquired pneumonia is complex, and a diagnostic algorithm is proposed. Appropriate samples are indicated during soft tissue and intraabdominal infections, but cultures obtained through the drains are discouraged. PMID- 15542958 TI - Antimicrobial therapy for patients with severe sepsis and septic shock: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for antimicrobial therapy for patients with severe sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Since the prompt institution of therapy that is active against the causative pathogen is one of the most important predictors of outcome, clinicians must establish a system for rapid administration of a rationally chosen drug or combination of drugs when sepsis or septic shock is suspected. The expanding number of antibacterial, antifungal, and antiviral agents available provides opportunities for effective empiric and specific therapy. However, to minimize the promotion of antimicrobial resistance and cost and to maximize efficacy, detailed knowledge of the likely pathogens and the properties of the available drugs is necessary for the intensivist. PMID- 15542959 TI - Source control in the management of severe sepsis and septic shock: an evidence based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for source control in the management of severe sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and to improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Source control represents a key component of success in therapy of sepsis. It includes drainage of infected fluids, debridement of infected soft tissues, removal of infected devices or foreign bodies, and finally, definite measures to correct anatomic derangement resulting in ongoing microbial contamination and to restore optimal function. Although highly logical, since source control is the best way to reduce quickly the bacterial inoculum, most recommendations are, however, graded as D or E due to the difficulty to perform appropriate randomized clinical trials in this respect. Appropriate source control should be part of the systematic checklist we have to keep in mind in setting up the therapeutic strategy in sepsis. PMID- 15542960 TI - Use of corticosteroid therapy in patients with sepsis and septic shock: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for the use of corticosteroid therapy in patients with sepsis and septic shock that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built upon a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSIONS: Low doses of corticosteroids are recommended in patients with septic shock. In the absence of vasopressor requirement, corticosteroids should not be used to treat sepsis. High-dose corticosteroids are not recommended in severe sepsis. The use of adrenal function tests to guide decisions on corticosteroid therapy, the weaning of steroids at the end of the treatment period, the decision to discontinue steroids earlier with resolution of shock, and the addition of oral fludrocortisone are considered optional approaches. PMID- 15542961 TI - Recombinant human activated protein C in the treatment of severe sepsis: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for recombinant human activated protein C that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Recombinant human activated protein C is recommended in patients at high risk of death (septic shock, sepsis-induced acute respiratory distress syndrome, Acute Physiology and Chronic Health Evaluation II score of >/=25, and sepsis-induced multiorgan failure) and no absolute contraindication related to bleeding risk or relative contraindication that outweighs the potential benefit. The presence or absence of disseminated intravascular coagulation should not influence the decision to administer recombinant human activated protein C. Heparin should be withheld during administration of recombinant human activated protein C. PMID- 15542962 TI - Use of blood products in sepsis: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for the use of blood products in sepsis that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and to improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: In the absence of extenuating circumstances and following resolution of tissue hypoperfusion, red blood cell transfusion should be targeted to maintain hemoglobin at 7.0 g/dL or greater. Erythropoietin is not recommended as a specific treatment for sepsis-associated anemia. Fresh frozen plasma should be given for documented deficiency of coagulation factors and in the presence of active bleeding or before surgical or invasive procedures. Antithrombin administration is not recommended. Specific platelet transfusion thresholds are based on the presence or absence of bleeding, significant risk for bleeding, plans for surgery or invasive procedures, and platelet count 7.15. PMID- 15542966 TI - Other supportive therapies in sepsis: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for other supportive therapies in sepsis that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Patients with severe sepsis should be treated with deep-vein thrombosis prophylaxis. Low-dose unfractionated heparin or low molecular weight heparin is preferred. Use of graduated compression devices is recommended in septic patients with contraindication to the use of heparin or combined with heparin in very high-risk patients. Stress ulcer prophylaxis should be given to all patients with severe sepsis. Histamine-2 receptor antagonists are more effective than sucralfate in decreasing bleeding risk and transfusion requirements. Proton pump inhibitors have not been assessed in a direct comparison with histamine-2 receptor antagonists but do demonstrate equivalency and ability to increase gastric pH. PMID- 15542967 TI - Sepsis associated with immunosuppressive medications: an evidence-based review. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for sepsis associated with immunosuppressive medications that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and improve outcome in severe sepsis. DESIGN: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. METHODS: The modified Delphi methodology used for grading recommendations built on a 2001 publication sponsored by the International Sepsis Forum. We undertook a systematic review of the literature graded along five levels to create recommendation grades from A to E, with A being the highest grade. Pediatric considerations to contrast adult and pediatric management are in the article by Parker et al. on p. S591. CONCLUSION: Immunosuppressed patients, by definition, are susceptible to a wider spectrum of infectious agents than immunologically normal patients and, thus, require a broader spectrum antimicrobial regimen when they present with sepsis or septic shock. Special expertise managing immunosuppressed patient populations is needed to predict and establish the correct diagnosis and to choose appropriate empiric and specific agents and maximize the likelihood that patients will survive these microbial challenges. PMID- 15542968 TI - Pediatric considerations. AB - OBJECTIVE: In 2003, critical care and infectious disease experts representing 11 international organizations developed management guidelines for other supportive therapies in sepsis that would be of practical use for the bedside clinician, under the auspices of the Surviving Sepsis Campaign, an international effort to increase awareness and to improve outcome in severe sepsis. DESIGN AND METHODS: The process included a modified Delphi method, a consensus conference, several subsequent smaller meetings of subgroups and key individuals, teleconferences, and electronic-based discussion among subgroups and among the entire committee. Pediatric representatives attended the various section meetings and workshops to contrast adult and pediatric management. These are published here as pediatric considerations. CONCLUSION: Pediatric considerations included a more likely need for intubation due to low functional residual capacity, more difficult intravenous access, fluid resuscitation based on weight with 40-60 mL kg or higher needed, decreased cardiac output and increased systemic vascular resistance as the most common hemodynamic profile, greater use of physical examination therapeutic endpoints, the unsettled issue of high-dose steroids for therapy of septic shock, and greater risk of hypoglycemia with aggressive glucose control. PMID- 15542969 TI - Sepsis change bundles: converting guidelines into meaningful change in behavior and clinical outcome. PMID- 15542971 TI - Update on Creutzfeldt-Jakob disease. AB - PURPOSE OF REVIEW: Prion diseases are transmissible fatal neurodegenerative disorders in which infectivity is associated with the accumulation of PrP(Sc), a disease-related isoform of normal cellular prion protein. The recent emergence of variant Creutzfeldt-Jakob disease has led to major public health concerns, and the need for the development of effective treatments. As PrP(Sc) is associated both with pathology and infectivity, therapeutic approaches to date have largely aimed at preventing its accumulation, but this strategy has produced only modest results in animal models. The link between PrP(Sc) and neurotoxicity is unclear, and alternative pathological processes need to be considered. Here we focus on the latest progress in therapeutic strategies and potential mechanisms of prion neurotoxicity. RECENT FINDINGS: Passive immunisation with anti-prion protein antibodies prevents peripheral prion replication and blocks progression to clinical disease in peripherally infected mice. A new approach, in which neuronal cellular prion protein is depleted in mice with established neuroinvasive prion infection, prevents the onset of clinical disease, blocks neuronal cell loss and reverses early spongiform pathology. This dramatic protective effect occurs despite the continued build-up of extraneuronal PrP(Sc) and continued replication of prion infectivity, effectively producing a sub-clinical state. SUMMARY: New insights into the mechanisms of neurotoxicity in prion diseases support the concept that PrP(Sc) itself is not directly neurotoxic. They suggest that neuronal prion propagation results in the production of a toxic intermediate or depletion of a key constituent. Prevention of the formation of such a species rather than PrP(Sc) accumulation itself is a clear target for prion therapeutics. PMID- 15542972 TI - Supportive management in neuro-oncology: opportunities for patient care, teaching, and research. PMID- 15542973 TI - Advances in the biology of astrocytomas. AB - PURPOSE OF REVIEW: Conventional surgery, radio- and chemotherapy have failed to significantly improve the prognosis of patients with malignant astrocytomas- hence the need for understanding their molecular biology. Harvesting this understanding to yield novel biological targeted therapies has approached the clinical doorstep. Therapeutic efficacy will likely require combinatorial therapy involving biologicals and conventional therapies, with small incremental efficacy in selected sub-groups. This review highlights some of the findings over the past year (June 2003-2004) that have contributed to this slow but essential journey towards our understanding of the biology of astrocytomas. RECENT FINDINGS: The accumulation of loss and/or gain of function molecular alterations underlying astrocytoma formation, progression and key growth parameters including proliferation, angiogenesis, apoptosis, invasion and resistance are emerging. These alterations involve those regulating the growth factor/receptor and downstream signaling networks, cell cycle, immune modulators and other key biological processes. The advances are facilitated by interactions amongst clinician and basic scientists, in both academia and industry. They have incorporated high-throughput bioinformatics analysis of genomic and expression array data, the emerging field of proteomics and development of various genetically engineered models of astrocytomas. SUMMARY: Astrocytomas, like other cancers, are a result of several molecular alterations, some of which strongly correlate to their pathological grade. However, molecular heterogeneity exists between astrocytomas of similar grades and likely between varying micro environmental regions of a single tumor. Characterization of the molecular signature of an astrocytoma and linking with the appropriate 'tailored' therapie(s) is the hope of the future. PMID- 15542974 TI - Non-cytotoxic drugs as potential treatments for gliomas. AB - PURPOSE OF REVIEW: Despite advances in surgery, radiation therapy, and chemotherapy, malignant gliomas continue to be associated with a poor prognosis. Even the most intensive combinations of radiotherapy and chemotherapy are not curative. In recent years our understanding of how tumor cells overcome cell cycle control, evade programmed cell death, induce blood vessel formation, and escape immune regulation has increased substantially. Significant efforts are directed towards the development of novel experimental therapies to target these molecular and biological mechanisms that lead to the development and growth of brain tumors. This review summarizes the most recent developments in non cytotoxic therapy for malignant gliomas, such as targeted molecular drugs, inhibitors of angiogenesis and intratumoral therapy. RECENT FINDINGS: The first generation of studies using these novel therapies is nearing completion. In general, most of these treatments are well tolerated, but single-agent activity is modest. There is significant interest in combining these therapies with each other and with conventional cytotoxic therapies such as radiation therapy and chemotherapy. SUMMARY: These new therapeutic approaches for malignant gliomas are showing modest activity. As we learn to use these agents more effectively, and as an increasing number of new and potentially promising agents are developed, it is likely that therapies for malignant gliomas will improve over the next few years. PMID- 15542975 TI - Advances in the biology and treatment of oligodendrogliomas. AB - PURPOSE OF REVIEW: The sensitivity of oligodendroglioma to chemotherapy and the prognostic significance of combined loss of 1p/19q in these tumors are now well established. This review discusses recent molecular, genetic and clinical advances made in studies on oligodendroglioma and mixed oligoastrocytoma. RECENT FINDINGS: Methylation of genes is a frequent event in oligodendroglioma (OD), but no specific methylation sites have been discovered. In contrast to earlier reports, the expression of the basic helix-loop-helix transcription factors is not specific for OD, but may occur in glial tumors of all lineages. In a number of studies, the prognostic relevance of 1p/19q loss has been confirmed, and several studies have shown that loss of 1p/19q are early events in OD. No candidate genes have so far been identified on 1p36 and 19q13. Gene expression profiling using gene arrays allows the separation of glial tumors according to histology, tumor grade and prognosis. A number of genes have been identified that are significantly more highly expressed in OD. On MRI imaging, OD with combined 1p/19q loss has typical characteristics, including indistinct borders and a mixed signal intensity on T2-weighted images. Despite the large increase in knowledge on the molecular abnormalities in OD, the therapeutic options for these tumors have not improved significantly since the introduction of temozolomide. The increased survival after chemotherapy has been clearly established, but the timing of chemotherapy seems less critical. It is clear that temozolomide is a good alternative to procarbazine, CCNU and vincristine (PCV) chemotherapy, in particular, because it is better tolerated. No randomized trials, however, have compared PCV-chemotherapy to temozolomide. New agents--and probably more targeted therapies--are needed to further improve treatment. Chemo-irradiation deserves further study in anaplastic OD. SUMMARY: The progress in the understanding of genetic and molecular abnormalities of OD has improved the recognition of treatment-sensitive OD, although this has not yet been mirrored in improved therapies or new treatment options. While chemotherapy improves the outcome of OD, further improvements will likely require new drugs or new treatment concepts. PMID- 15542976 TI - Update of stereotactic radiosurgery for brain tumors. AB - PURPOSE OF REVIEW: This paper will review the recent publications of stereotactic radiosurgery for brain tumors. RECENT ADVANCES: Despite its controversial beginning, stereotactic radiosurgery has rapidly gained acceptance among neurosurgeons, radiation oncologists, and neuro-oncologists as a valuable treatment option for patients with certain benign and malignant brain tumors. Over the past year, a number of publications have confirmed the efficacy and safety of this treatment modality as the sole treatment modality or as part of the multimodality management of brain tumor patients. These publications ranged from the first multi-institutional phase III trial of radiosurgery for patients with brain metastases to numerous retrospective papers about treatment outcomes. Also, a number of these publications have explored the use of newer imaging modalities to improve treatment outcomes while others reported on the rare complication of radiation-associated second tumors. SUMMARY: Recent publications of stereotactic radiosurgery have increased our understanding of the use of this technology. Future studies are needed to further improve outcomes, minimize toxicities and increase our understanding of this treatment modality. PMID- 15542977 TI - Meningioma: an update. AB - PURPOSE OF REVIEW: Recent clinical and molecular research has shed new light on the biology of meningiomas--a common but understudied CNS neoplasm. This review will focus on recent advances and their significance for future research and treatment. RECENT FINDINGS: Meningiomas represent the second most common brain tumor in adults, and while improved diagnostic modalities are available, these tumors remain underreported. Radiosurgery is an effective adjuvant therapy against meningioma; however, no effective chemotherapy exists. In addition to histologic grading and estimates of the extent of resection, biomarkers, such as progesterone receptor, cyclooxygenase 2, S100A5 and ornithine decarboxylase may be useful in predicting tumor recurrence and/or progression potential in patients with meningioma. On the genetic level, cytogenetic losses on chromosomes 1, 7, 10 and 14 and telomerase activation are observed in clinically aggressive meningioma, whereas monosomy 22 is a common early molecular event in tumor formation. Several candidate growth regulatory genes have been identified, including the Neurofibromatosis 2 (NF2), Tumor Suppressor in Lung Cancer-1 (TSLC1), Protein 4.1B, p53/MDM2 and S6-Kinase genes. The roles of these genes in meningioma formation and progression, as well as the clinical implications of these genetic changes, are discussed. SUMMARY: The recent insights into the molecular biology and genetics of meningioma provide new avenues for basic science research aimed at understanding the mechanisms underlying meningioma formation and malignant progression. These advances may be useful in improving our ability to predict clinical outcome and developing targeted therapies to improve outcomes in patients with clinically aggressive meningiomas. PMID- 15542978 TI - Diagnosis and treatment of pituitary tumors. AB - PURPOSE OF REVIEW: Treatment of pituitary adenomas remains an interdisciplinary challenge involving neurosurgeons, endocrinologists and radiation oncologists. The different disciplines inaugurated advanced techniques to improve the already relatively high standard of outcome for the benefit of patients, covering molecular pathogenesis, novel therapeutic strategies for the different adenoma subtypes, developments in perioperative magnetic resonance imaging and radiosurgical management of pituitary adenomas. RECENT FINDINGS: Despite the progress achieved in medical treatment of hormone-secreting pituitary adenomas throughout recent years, surgery remains the primary therapy of choice except for prolactinomas. Recent studies in molecular pathogenesis aiming to find novel therapy targets and reports on new pharmacological drugs effecting GH-secreting pituitary adenomas are reviewed (for example, lanreotide 60, SOM320 and pegvisomant). Advances in surgical treatment of pituitary macroadenomas are obtained by pre- and especially by intraoperative (high-field) MRI offering a higher rate of safe and complete tumor removal. Therapy pitfalls mentioned in the literature throughout the last year as well as key points in the management of pituitary adenomas with focus on acromegaly and Cushing's disease are reported. Adjuvant irradiation for recurrent or residual adenomas is often a necessity. In comparison to standard conventional radiation strategies an increasing number of radiation oncologists and neurosurgeons report their experience with radiosurgery especially for smaller tumor remnants in pituitary adenomas. SUMMARY: Recent molecular studies suggest a new level of complexity in the tumorigenisis of pituitary adenomas in terms of possible cell-type-specific molecular changes. Except for prolactinomas surgery remains the primary treatment for pituitary adenomas. New pharmacological drugs achieve very encouraging endocrine results although no long-term follow-up is available so far. The results of trans sphenoidal surgery will further improve by modern imaging techniques, especially by applying intraoperative high-field magnetic resonance imaging and neuronavigation. The results of radiosurgical techniques with regard to tumor control are mostly convincing, but definitive conclusions on long-term recurrence and/or late complications are not reliable so far. PMID- 15542979 TI - The functional relevance of cerebral hemodynamics: why blood flow matters to the injured and recovering brain. PMID- 15542980 TI - Traumatic brain injury: physiology, mechanisms, and outcome. AB - PURPOSE OF REVIEW: This review on traumatic brain injury consolidates the substantial current literature available on the pathophysiology, mechanisms, developments, and their subsequent effects on outcome. In particular, it tries to conceptualize why our greatly improved understanding of pathophysiology and neurobiology in traumatic brain injury has not translated into clear outcome improvements. RECENT FINDINGS: Early cerebral ischaemia has been characterized further, with ischaemic brain volume correlating with 6-month outcome. The Brain Trauma Foundation has revised perfusion pressure targets, and there are additional data on the outcome impact of brain tissue oxygen response and asymmetric patterns of cerebral autoregulation. Mechanistic studies have highlighted the role of inflammation and introduced concepts such as therapeutic vaccination and immune modulation. Experimental neurogenesis and repair strategies show promise. Despite continuing gains in knowledge, the experimental successes have not yet translated to the clinic. Indeed, several major articles have attempted to understand the clinical failure of highly promising strategies such as hypothermia, and set out the framework for further studies (e.g. addressing decompressive craniectomy). High-dose mannitol has shown promise in poor grade patients, while hypertonic saline has shown better intracranial pressure control. Negative results may be the consequence of ineffective therapies. However, there is a gathering body of work that highlights the outcome impact of subtle neurocognitive changes, which may not be quantified adequately by outcome measures used in previous trials. Such knowledge has also informed improved definition of mild traumatic brain injury, and allowed validation of management guidelines. SUMMARY: The evidence base for current therapies in this heterogeneous patient group is being refined, with greater emphasis on long-term functional outcomes. Improved monitoring techniques emphasize the need for individualization of therapeutic interventions. PMID- 15542981 TI - Head injury and dementia. AB - PURPOSE OF REVIEW: The link between head injury and dementia/Alzheimer's disease is controversial. This review discusses some recent epidemiological, human autopsy and experimental studies on the relationship between traumatic head injury and dementia. RECENT FINDINGS: Recent epidemiological studies have shown that head injury is a risk factor for the development of dementia/Alzheimer's disease, whereas others have not. After experimental brain trauma the long-term accumulation of amyloid beta peptide suggests that neurodegeneration is influenced by apolipoprotein E epsilon 4, and after human brain injury both amyloid beta peptide deposition and tau pathology are seen, even in younger patients. Amyloid beta peptide levels in the cerebrospinal fluid and the overproduction of beta amyloid precursor protein in humans and animals after traumatic brain injury are increased. Repeated mild head trauma in both animals and humans accelerates amyloid beta peptide accumulation and cognitive impairment. Retrospective autopsy data support clinical studies suggesting that severe traumatic brain injury with long-lasting morphological residuals are a risk factor for the development of dementia/Alzheimer's disease. The influence of the apolipoprotein E genotype on the prognosis of traumatic brain injury is under discussion. SUMMARY: Although epidemiological studies and retrospective autopsy data provide evidence that a later cognitive decline may occur after severe traumatic brain injury, the relationship between dementia after head/brain trauma and apolipoprotein E status is still ambiguous. Both human postmortem and experimental studies showing apolipoprotein beta deposition and tau pathology after head injury support the link between traumatic brain injury and dementia, and further studies are warranted to clarify this relationship. PMID- 15542982 TI - Functional reorganization of the cerebral motor system after stroke. AB - PURPOSE OF REVIEW: Recovery of function after stroke is now widely considered to be a consequence of central nervous system reorganization. Non-invasive techniques such as functional magnetic resonance imaging, transcranial magnetic stimulation, electroencephalography and magnetoencephalography now allow the study of the working human brain. Studies in stroke patients can now address how cerebral networks in the human brain respond to focal injury and whether these changes are related to functional recovery. This understanding may in turn lead to the development of techniques that will drive cerebral reorganization in a way that promotes functional improvement. RECENT FINDINGS: The relationship between cerebral reorganization and functional recovery has been examined in both cross sectional and longitudinal studies. It appears that the motor system reacts to damage in a way that attempts to generate motor output through surviving brain regions and networks. There are changes in cortical excitability after stroke that may provide the substrate whereby the effects of motor practice or experience can be more effective in driving long lasting changes in motor networks. This will be particularly important in intact portions of neural networks subserving motor skills learning. SUMMARY: Functionally relevant adaptive changes occur in the human brain following focal damage. A greater understanding of how these changes are related to the recovery process will allow the development of novel therapeutic techniques that are based on neurobiological principles and which are designed to minimize impairment in appropriately targeted patients suffering from stroke. PMID- 15542984 TI - Bibliography. Current world literature. Degenerative and cognitive diseases. PMID- 15542983 TI - Epilepsy after head injury. AB - PURPOSE OF REVIEW: The purpose of this short review is to provide an update on the epidemiology of posttraumatic epilepsy, associated risk factors, data from prevention studies, and recent breakthroughs in experimental research. RECENT FINDINGS: There is increasing evidence that neuroimaging findings, stratification by neurosurgical procedures performed, and genomic information (e.g. apolipoprotein E and haptoglobin genotypes) may provide useful predictors of the individual risk of developing posttraumatic epilepsy. While antiepileptic drug prophylaxis can be effective in protecting against acute (provoked) seizures occurring within 7 days after injury, no antiepileptic drug treatment has been found to protect against the development of posttraumatic epilepsy and therefore long-term anticonvulsant prophylaxis is not recommended. Glucocorticoid administration early after head injury also has not been found to reduce the risk of posttraumatic epilepsy. At the basic research level, there have been advances in the understanding of pathophysiological changes in posttraumatic excitatory and inhibitory synapses, and the critical period for epileptogenesis after head injury has been better defined. Finally, the development of a novel animal model, which mimicks more closely human posttraumatic epilepsy, may facilitate efforts to characterize relevant epileptogenic mechanisms and to identify clinically effective antiepileptogenic treatments. SUMMARY: Despite the continuing lack of clinically effective agents for posttraumatic epilepsy prophylaxis, recent advances in basic and clinical research offer new hope for success in the development of new strategies for prevention and treatment. PMID- 15542986 TI - Administration of analgesia for acute abdominal pain sufferers in the accident and emergency setting. AB - BACKGROUND: Recent trials suggest that the early administration of analgesia in patients with acute abdominal pain facilitates examination and does not delay diagnosis. We investigated current practice regarding analgesia for these patients. METHODS: All patients admitted via the accident and emergency department with abdominal pain were included. The main outcome measures evaluated were waiting time for analgesia and its relationship to subjective visual analogue pain scores and clinical diagnoses. RESULTS: Data from 107 consecutive patients were investigated; seven patients were excluded. Forty-two per cent were male. The mean age was 40.1 years (6-85). The mean overall waiting time for analgesia was 1.4 h (2 min to 14 h). Sixty-seven per cent received analgesia within one hour, although 22% waited 2-14 h after presentation. Those with mild pain waited significantly longer for analgesia (mean 247 min) than those with severe pain (mean 82 min; P=0.01). Those with moderate pain had intermediate waiting times (mean 111 min), although they were not statistically different from the severe group (P=0.43). Female patients had to wait longer (mean 129 min) than male patients (mean 69 min; P=0.09 analysis of variance). Of 64% who were general practitioner referrals, only 11% (all severe group, P=0.02) received analgesia in the community. Neither clinical diagnosis nor age influenced the timing of analgesia. Seventy-three per cent received analgesia in the casualty department (mean 0.5 h; range 0.02-3.2), whereas those admitted in the ward without receiving analgesia in casualty had to wait significantly longer for their pain relief (mean 5 h; 1.2-14). CONCLUSION: This study shows the need for standardized protocols for analgesia usage in patients presenting with acute abdominal pain. PMID- 15542987 TI - Is the management of dog bite wounds evidence based? A postal survey and review of the literature. AB - OBJECTIVES: To determine current practice in the management of dog bite wounds with regard to the use of prophylactic antibiotics and primary closure and to compare the available evidence. METHODS: We conducted a national postal survey of Accident and Emergency (A and E) departments in the UK to ascertain the current practice in the management of recent dog bite wounds. A questionnaire was designed and posted to a named A and E consultant. Non-respondents were sent a single reminder 8 weeks after the initial mailing. A detailed literature search was carried out using Medline, Embase and Cochrane databases to look at the available evidence. RESULTS: The postal survey yielded an 80% response. Prophylactic antibiotics were routinely used by 53% of the respondents. Although 99% of the respondents were happy to close facial dog bite lacerations, 60% were reluctant to suture other body regions. Recent available evidence suggests that restricting the use of prophylactic antibiotics for high-risk dog bite wounds and the primary suturing of appropriate dog bite lacerations is safe. CONCLUSIONS: The management of recent dog bite wounds is not entirely evidence based in many A and E departments in the UK. PMID- 15542988 TI - Drug and alcohol use among patients admitted to a Danish trauma centre: a prospective study from a regional trauma centre in Scandinavia. AB - OBJECTIVES: The incidence of drug and alcohol use is unknown among Danish trauma patients, and has not been thoroughly investigated in Europe. METHODS: Patients admitted to the regional trauma centre in Aarhus, Denmark, were prospectively screened by blood and urine tests for the presence of alcohol, and legal and illicit drugs. The correlation with the Injury Severity Score, hospitalization time, and mortality after drug or alcohol intake was investigated. RESULTS: A quarter of all patients admitted in 1999 and 2000 had an alcohol level exceeding the national legal driving limit of 50 mg/dl, and one or more drugs were found in one in five patients. The presence of any drug or alcohol correlated positively with the Injury Severity Score, whereas alcohol level, hospitalization time, and mortality did not correlate. CONCLUSION: The rate of trauma patients with a blood serum level of alcohol greater than 100 mg/dl was similar to a previous European study and to US studies. The prevalence of drugs was less, and benzodiazepines were the most used group of drugs in our study. The routine screening of Danish trauma patients in order to implement preventative measures may be beneficial. PMID- 15542989 TI - US emergency department visits for hip fracture, 1992-2000. AB - BACKGROUND: Hip fracture is a significant injury for older persons. Little has been reported about emergency department (ED) care of these patients. OBJECTIVE: To characterize the management of older patients treated in the ED for hip fracture with specific attention to the use of analgesia. METHODS: This study analysed data from the National Hospital Ambulatory Medical Care Survey ( approximately 400 hospitals report each year) for the years 1992-2000. We included records of patients who were at least 50 years old and had a diagnosis of hip fracture (International Classification of Disease, 9th revision, clinical modification 820-820.9) in any of the three fields allowed for recording diagnosis. RESULTS: There were 1,935,000 ED visits ( approximately 215,000 a year), predominantly among older white women. The rate in those over 80 years old was 25 times that of the youngest group. Fifty-six per cent of patients received analgesia (44% narcotics). There were no ethnic or racial distinctions in the use of analgesia. CONCLUSIONS: The ED visit presents a largely untapped opportunity for focused efforts in fall and hip fracture prevention, especially in countries with comprehensive geriatric services and well integrated healthcare delivery systems. Despite the high likelihood of pain, the administration of analgesics appears to be low and may be a worthy focus for practice improvement. PMID- 15542990 TI - Functions and principles in the management of bombing mass casualty incidents: lessons learned at the Tel-Aviv Souraski Medical Center. AB - Mass casualty incidents (MCI) resulting from terrorist bombings pose special problems and may overwhelm even the most experienced trauma centre. Although role assignments for MCI management and control are documented, we would like to share several aspects that we have found to be crucial for the management of terrorist bombing MCI. Twelve topics and possible answers were brought up and discussed extensively in MCI debriefing sessions in our institution. They were of two main categories: assignments and functions of key personnel, and principles and definitions. The first category includes the Triage Officer, the Medical Director, the Administrative Director, the Head Nurse, the Emergency Medical System Coordinator, the Blood Bank Liaison, and the trauma teams. The second category encompasses the concept of triage hospital, the unidirectional patient flow, ancillary evaluation during MCI, the consultancy, and tertiary survey. All were identified as critical for proper event handling. The integration and implementation of the topics discussed throughout the medical system may enable emergency departments to handle MCI resulting from terrorist bombings better. PMID- 15542991 TI - Resuscitative mild hypothermia as a protective tool in brain damage: is there evidence? AB - Resuscitative mild hypothermia is and will increasingly be used in the emergency department as protection for the brain after an ischaemic insult. The clinical application of resuscitative mild hypothermia and its limitations will be summarized in this paper. The evidence for each application and its underlying mechanism will also be reviewed. PMID- 15542992 TI - A case of multiple missed fractures! PMID- 15542993 TI - Pulmonary thromboembolism presenting as epileptiform generalized seizure. AB - A 69-year-old patient with a new-onset generalized epileptiform tonic-clonic seizure was successfully diagnosed with pulmonary thromboembolism in the prehospital setting even though the clinical picture suggested an unknown, primarily neurological, problem. Prehospital diagnostic procedures also included capnometry and D-dimer assay. The former showed lower than normal end-tidal carbon dioxide pressure values, and a value of the latter proved to be highly pathological. In the intensive care unit the diagnosis of massive pulmonary thromboembolism was confirmed. PMID- 15542994 TI - Bilateral pleural effusion and ascites in the ovarian hyperstimulation syndrome. AB - We describe a pregnant woman with ovarian hyperstimulation syndrome with bilateral pleural effusion and ascites. Ovarian hyperstimulation syndrome is an iatrogenic complication of ovarian stimulation, characterized by a massive crossing of a protein-rich fluid from the vascular compartment into the peritoneal, pleural, or to a lesser extent, pericardial cavities. Management is usually conservative, with fluid and electrolytes correction and thromboprophylaxis. Prevention is very difficult, but an age younger than 35 years, low body mass index, polycystic ovarian disease, a high number of follicles, a high plasma oestradiol concentration, pregnancy, hyperandrogenism, and hypothyroidism are predisposing factors. PMID- 15542995 TI - Expect the unexpected: malposition of a large-bore central venous catheter in the urinary bladder. AB - We report the case of a femoral vein cannulation in a critically ill trauma patient with the malposition of a large-bore central venous catheter in the urinary bladder. Recognition of the malposition was hampered by bloody tamponade of the bladder in the context of blunt thoraco-abdominal trauma with kidney and liver laceration. A high index of clinical suspicion and the institution of adequate therapy were the key to achieving a successful clinical outcome. We discuss the anatomy of femoral veins, including their close relation to a distended bladder. The application of ultrasound even in emergency situations is stressed. PMID- 15542996 TI - Hypercapnia and oxygen therapy in older asthmatic patients. AB - Excessive oxygen administration in hypercapnic chronic obstructive pulmonary disease predisposes to worsening respiratory failure during intercurrent respiratory illness. Chronic hypercapnia is thought to downregulate carbon dioxide chemoreceptor sensitivity, adversely affecting respiratory function/mechanics and worsening ventilation-perfusion inequality. These patients are dependent on hypoxic drive to maintain adequate spontaneous respiration. Whether an analogous situation occurs in asthma in older adults is unknown. These conditions may be difficult to differentiate clinically, and both may respond adversely to the administration of excessive oxygen in the presence of chronic hypercapnia. Although unrestricted oxygen is beneficial and safe in children and young adults with asthma, it may lead to progressive hypercapnia in older patients with asthma, a potential risk highlighted by this case. To avert progressive hypercapnia, oxygen therapy that is carefully adjusted to achieve adequate, but not maximal, tissue oxygenation may be a safer strategy than unrestricted oxygen use in older asthmatic patients. However, the correction of hypoxia overrides strategies to avert oxygen-related hypercapnia. PMID- 15542997 TI - Spinning-induced rhabdomyolysis: a case report. AB - Exercise induced rhabdomyolysis has previously been described as occurring in unfit adults or those attempting exhaustive exercise, but is rarely seen in previously fit adults undertaking moderate exercise. This case report describes what we believe to be the first reported case of exercise induced rhabdomyolysis due to the patient completing a 'spinning class' at her local gymnasium. The report also covers diagnosis and management of this condition. PMID- 15542998 TI - Exacerbation of atopic dermatitis in the emergency department. AB - A 38-year-old man was admitted to the Emergency Department suffering from an exacerbation of atopic dermatitis, fever and a burning sensation in the eyes. He was first treated with systemic corticosteroids. A subsequent dermatological and ophthalmological examination established the diagnosis of Kaposi-Juliusberg disease or eczema herpeticum with bilateral herpetic keratitis. Eczema herpeticum is an uncommon herpes simplex virus infection that occurs in patients with atopic dermatitis. Because it is a possible life-threatening condition, this disease must be recognized by all emergency physicians. The association with herpetic keratitis is not frequent but is a major ophthalmological problem. Treatment consists of the administration of high-dose intravenous acyclovir and acyclovir ophthalmic ointment. PMID- 15542999 TI - Coeliac crisis in adults: report of two cases. AB - Adult coeliac disease, in contrast to its childhood counterpart, almost always has an indolent course with a wider spectrum of clinical manifestations. Approximately half of the patients have no overt gastrointestinal symptoms and many are asymptomatic. A rare and life-threatening complication, affecting mainly children younger than 2 years of age, is the so-called coeliac crisis, a term that applies to profuse diarrhoea leading to dehydration, hypokalemia, and acidosis. We report here two cases of adult coeliac disease that presented as coeliac crisis. Coeliac disease should be a differential diagnosis in adult patients with severe acute diarrhoea and acidosis, although a rare presentation. PMID- 15543000 TI - What is the price of an erythrocyte and neutrophil? PMID- 15543001 TI - Predictive value of childhood cancer diagnoses in the Danish Cancer Registry. PMID- 15543002 TI - Favorable-biology neuroblastoma presenting with leptomeningeal metastases?: a case presentation. AB - Intrinsic biologic tumor features are critical prognosticators of survival in patients with neuroblastoma. Patients with localized neuroblastoma and favorable biologic parameters may be observed without treatment. Conversely, leptomeningeal metastases in patients with primary extracranial neuroblastoma are highly unusual and, despite aggressive multimodality therapies, invariably fatal. The authors describe a newly diagnosed infant with neuroblastoma and radiographic imaging suggestive of leptomeningeal metastases. The patient underwent partial surgical resection of the primary tumor. The primary tumor revealed favorable biologic characteristics. The patient was observed with no cytotoxic therapy and remained well with no evidence of disease progression more than 3 years since diagnosis. This case illustrates that some infants with favorable-biology neuroblastoma may be observed without treatment despite the advanced INSS stage. PMID- 15543003 TI - Tumor necrosis factor, interleukin 11, and leukemia inhibitory factor produced by Langerhans cells in Langerhans cell histiocytosis. AB - OBJECTIVES: : The etiology and pathophysiology of Langerhans cell histiocytosis (LCH) remain elusive. The 3-year survival in pediatric multisystem LCH is still around 80%, and children with risk organ involvement (i.e., liver, spleen, hematopoietic system, or lungs) have a less favorable outcome. To further elucidate the pathogenesis of LCH in the search for a rationale cure, the authors investigated intracellular synthesis of tumor necrosis factor (TNF), interleukin (IL)-11, and leukemia inhibitory factor (LIF) from biopsied lesions. METHODS: : Lesional cells were obtained by fine-needle aspiration biopsy from nine children with LCH. The study was accomplished by the use of an immunofluorescence staining method that allowed cytokine-producing cells to be differentiated from cytokine binding cells. RESULTS: : All patients had histiocytes expressing TNF. Seven patients had histiocytes expressing IL-11 and six patients had histiocytes expressing LIF. The two children with the highest proportion of histiocytes displaying TNF and the three with the highest proportion of histiocytes expressing IL-11 and LIF all had risk organ involvement. Two-color staining revealed that histiocytes expressing TNF, IL-11, and LIF co-expressed CD1a molecules. CONCLUSIONS: : These observations suggest that LCH represents a cytokine-driven condition partially mediated by TNF, IL-11, and LIF. These three cytokines are all osteoclastogenic, suggesting a pathogenetic pathway for the osteolytic lesions in LCH. Furthermore, thrombocytosis in LCH may be explained by IL-11 and LIF activity. PMID- 15543004 TI - Massive congenital intracranial teratoma: perioperative coagulation impairment. AB - Massive congenital intracranial teratoma is a rare neoplasm with a poor prognosis. Surgery may be curative only if complete resection can ben obtained. Several single case reports have been published in the pediatric literature, mostly focusing on prenatal diagnosis. The authors describe six patients with congenital intracranial teratoma treated at their institution in the past decade. Perioperatively, a marked hemostatic derangement was observed in three of them undergoing surgery: the pathophysiology of this complication is discussed. The surgical indication itself represents an ethical dilemma when treating a large intracranial tumor with partial destruction and replacement of brain structures. PMID- 15543005 TI - Immunophenotype of blood lymphocytes in neuroblastoma-associated opsoclonus myoclonus. AB - OBJECTIVE: To determine whether the distribution of peripheral blood mononuclear cells (PBMCs) is altered in paraneoplastic opsoclonus-myoclonus (POM). METHODS: PBMCs from 17 children with POM, 17 children with OM but no tumor, and 17 controls were immunophenotyped using a comprehensive panel of surface markers by dual-laser flow cytometry. All groups were matched for age and gender; POM and OM patients were matched for treatment. RESULTS: In the POM patients, the CD4+ T cell subset was smaller in both relative size (-18%, P = 0.02) and absolute size (-41%, P = 0.03) compared with controls. The CD4/CD8 ratio also was less (-29% to -44%) and was related to POM duration (P = 0.03). The absolute but not relative size of the gammadelta T-cell subset was reduced (-44%, P = 0.02). There were no significant abnormalities of CD19+ B-cells, CD3- or CD3+ NK cells, HLA-DR+ or CD25+ T-cells, or CD45RA+ or CD45RO+ T-cells. Prior tumor chemotherapy, which was associated with a higher percentage but not number of CD8+ T-cells, did not restore the CD4+ T-cell subset. When the POM and OM groups, which were not significantly different, were combined, chemotherapy decreased both the relative and absolute size of the CD19+ B-cell pool and had small effects on other lymphocyte subsets. CONCLUSIONS: POM is characterized by T-cell abnormalities of PBMCs, the most robust of which is reduction of the CD4+ T-cell subset and the CD4/CD8 ratio. Although this reduction was found previously in cerebrospinal fluid in POM patients, PBMC subsets did not otherwise reflect cerebrospinal fluid abnormalities. Longitudinal studies will be necessary to determine whether PBMC abnormalities could serve as treatment markers. PMID- 15543006 TI - Long-term outcome of chronic idiopathic thrombocytopenic purpura in children. AB - OBJECTIVES: Children with chronic idiopathic thrombocytopenic purpura (ITP) generally have a favorable outcome, but it is not known whether there are any prognostic factors to predict outcome. The objectives of this study were to assess the spontaneous remission rate and the prognostic significance of age, gender, initial platelet count, initial treatment, and response to treatment. METHODS: In this retrospective review of 62 consecutive children with chronic ITP, 37 were girls and 27 were 10 years of age or older (median age 9 years; range, 0.75-19). RESULTS: Thirty-five patients (56%) achieved spontaneous remission (remission without splenectomy), 30 of them (48%) within 4 years from diagnosis. Twenty-eight (45%) were complete remissions (platelet counts of >/=100,000) and 7 (11%) were partial remissions (50,000-99,000). There was no significant difference in the spontaneous remission rate between the younger (<10 years) and older children (55.8% vs. 57.1%, P = 0.95) or between boys and girls (56% vs. 56.7%, P = 0.98). Similarly, platelet count at initial diagnosis, initial therapy, or response to initial therapy did not have any prognostic significance. All 14 patients who underwent splenectomy achieved complete remission. CONCLUSIONS: More than 50% of children with chronic ITP achieve spontaneous remission. Age, gender, platelet count at initial diagnosis, initial treatment, and response to initial treatment do not have any prognostic significance toward the outcome of chronic ITP. PMID- 15543007 TI - Intensive treatment for childhood acute lymphoblastic leukemia reduces immune responses to diphtheria, tetanus, and Haemophilus influenzae type b. AB - OBJECTIVES: Immunity to diphtheria toxoid (D), tetanus toxoid (T), and Haemophilus influenzae type b (Hib) is affected in children with acute lymphoblastic leukemia (ALL). The aims were to examine immunity and to compare the response to immunization at 1 or 6 months after treatment. METHODS: Thirty one patients were immunized with DT and conjugated Hib vaccine (ActHib) at 1 month or 6 months after treatment of ALL with the NOPHO 92 protocol. Antibody levels were determined before and 3 weeks after vaccination. Specific T and Hib antibody-secreting cells of IgG/IgA/IgM isotypes were analyzed in peripheral blood using an ELISPOT technique. RESULTS: All specific antibody levels decreased during ALL treatment, and protective levels after treatment were noted for 17% against D, 33% against T, and 100% against Hib. No high-risk patient had full D or T protection after treatment. After vaccination all the standard- and intermediate-risk patients achieved full protection against D, T, and Hib. The high-risk group showed insufficient immune response (full protection after vaccination: D 56%, T 22%, Hib 78%). No difference was found between vaccination at 1 month or 6 months after treatment. The poor antibody production in the high risk group correlated to low numbers of antibody-secreting cells. CONCLUSIONS: Nonprotective antibody levels against D, T, and Hib after childhood ALL are more common than previously thought. Insufficient immune response was restricted to the high-risk group and was related to a low number of memory B cells in this study. Immunizations should be included in follow-up after childhood ALL, and the policy should be adapted to treatment intensity. PMID- 15543008 TI - Cerebellar ataxia in pediatric patients with Langerhans cell histiocytosis. AB - The pathogenetic mechanisms of the central nervous system (CNS) problems associated with Langerhans cell histiocytosis (LCH) are not well established. Effective treatment strategies for these CNS complications are not yet available, while diabetes insipidus, also associated with LCH, can be managed effectively. Three Japanese boys with LCH who developed cerebellar ataxia were evaluated. Similar pediatric cases from the literature are also discussed. All three patients initially developed multifocal LCH lesions during early childhood (age <3 years) that responded well to chemotherapy; however, two of the three patients later developed diabetes insipidus. Ataxia, associated with mild developmental delay, was noted in the patients between the ages of 4 to 8 years. Analysis of these three cases, along with previously reported cases, indicates that the median age of onset of LCH was 2.5 (range 0.1-6.5) years and the median age of onset of cerebellar lesions/ataxia was 7 (range 3.5-16.5) years. Although the incidence of cerebellar LCH involvement is low, delayed onset of CNS disease must be monitored during follow-up care of pediatric LCH patients. Brain magnetic resonance imaging is strongly recommended for early detection of cerebellar lesions, but it remains to be determined whether there are any therapeutic measures to prevent exacerbation of CNS disease. PMID- 15543009 TI - Outcome after autologous hemopoietic stem cell transplantation in relapsed or refractory childhood Hodgkin disease. AB - OBJECTIVES: To determine the clinical outcome and prognostic factors for overall survival in children with recurrent and/or primary refractory Hodgkin disease (HD) after high-dose therapy and autologous hemopoietic stem cell transplantation (AHSCT). The survival outcome of this treatment was compared with conventional salvage therapy without stem cell transplantation. METHODS: Clinical records of 51 patients with relapsed or refractory HD who underwent AHSCT were reviewed. The source of the stem cells was bone marrow (n = 22) or peripheral blood (n = 29). At the time of high-dose therapy, 39 patients were in complete remission and 1 was in partial remission, while the remaining 11 had refractory disease. The records of 78 patients from the HD 1 trial who underwent conventional salvage treatment but without AHSCT for relapsed or refractory HD were also reviewed. All patients received HDT without radiation for conditioning. RESULTS: Overall survival from diagnosis of patients treated with AHSCT did not differ significantly from that of those treated with conventional salvage therapy (hazard ratio = 1.5; 95% confidence interval = 0.9-8.2; P = 0.4). There were also no statistically significant differences in survival data between the two approaches for patients whose duration of first remission was less than or greater than 1 year (P = 0.5; stratified log-rank). Of the 11 patients who received AHSCT for refractory disease, 9 remain alive and well with followups ranging from 2 to 18 years. No deaths due to treatment-related complications were seen in the AHSCT group. CONCLUSIONS: Stem cell transplantation does not offer any significant survival advantage over conventional salvage therapy in children with relapsed HD, although it may be of benefit for patients with primary refractory disease. PMID- 15543010 TI - A comprehensive study of the neonatal manifestations of congenital dyserythropoietic anemia type I. AB - Congenital dyserythropoietic anemia (CDA) type I is an inherited disorder characterized by macrocytic anemia with pathognomonic morphologic ultrastructural features of the erythroid precursors. The authors recently cloned the CDAN1 gene and identified one founder missense mutation in all of their Bedouin patients. In a previous study, the authors found that the majority of their 31 Bedouin patients had anemia and jaundice during the first month of life and required blood transfusions; some had persistent pulmonary hypertension. In the present retrospective evaluation of 70 Bedouin patients with CDA type I, the authors more than doubled the number. Forty-five (64%) patients were symptomatic in the neonatal period, 29 (65%) had hepatomegaly, 24 (53%) had early jaundice, 11 (27%) were born small for gestational age, 7 (15%) had persistent pulmonary hypertension, and 6 (13%) had direct hyperbilirubinemia and another 6 (13%) had transient thrombocytopenia. Thirty-six of the symptomatic neonates (80%) required at least one blood transfusion. These results confirm the authors' previous findings and add neonatal manifestations not previously described, particularly hyperbilirubinemia and thrombocytopenia. Early diagnosis of CDA type I may be beneficial in light of the potential efficacy of alpha-interferon in avoiding transfusions in some patients. PMID- 15543011 TI - Clinical presentation and treatment outcome of children with Burkitt lymphoma in Lebanon: a single institution's experience. AB - The authors reviewed the medical records of 42 children younger than 13 years of age diagnosed with Burkitt lymphoma at the American University of Beirut Medical Center between 1983 and 1993. The male:female ratio was 3.9. The abdomen was the most common site of disease (86%). Jaw, central nervous system, and bone marrow involvement occurred in 16.6%, 16.6%, and 9.5%, respectively. The mean LDH level was 447 U/L. The mean age at diagnosis was 6.9 years. Thirty-nine patients received a variation of the COMP protocol. The total duration of treatment ranged from 6 to 18 months. At a median follow-up of 5 years the event-free survival was 100% for children with stages I and II disease, 77.4% (+/- 2 SE) for stage III, and 0% for stage IV. Failures in stage III patients were due to tumor lysis (3/24) and progressive disease (2/24). Aggressive therapy with high doses of methotrexate and anthracyclines may not be necessary for the treatment of children with extensive abdominal disease (stage III) in Lebanon. If confirmed in a larger series of patients, this study could have a major impact on the treatment of Burkitt lymphoma in Lebanon and other countries with limited resources. PMID- 15543012 TI - Posttraumatic intracerebral venous thrombosis: an infant who has fallen off a hammock. AB - A 45-day-old boy presented with drowsiness, pallor, and respiratory failure of about 24 hours' duration. Initially, the family gave no history of trauma, but after the laboratory investigations, they admitted that he had fallen off a hammock. On the plain films and cranial computed tomography (CT), no fracture of the cranium was detected, but cranial CT and magnetic resonance imaging revealed thrombosis in the sigmoid sinus, superior sagittal sinus, and straight sinus; right subdural hematoma; diffuse cerebral edema; and diffuse infarction in the right posterior temporal, parietal, and occipital lobes. Full recovery was achieved by anticoagulation with unfractionated heparin within 14 days. Falling off a hammock was the only risk factor for this patient. Thus, when cerebral thrombosis develops in childhood, trauma should be considered regardless of whether such information is provided by the family. PMID- 15543013 TI - Carboplatinum and vincristine chemotherapy for central nervous system gliofibroma: case report and review of the literature. AB - Gliofibromas are bimorphic tumors of the central nervous system. Although they are composed of astrocytic and fibroblastic elements, their histogenesis is not clear. An attempt has been made to classify the tumors as low- or high-grade based on morphology and proliferative labeling index, but the clinical behavior and the optimal therapeutic strategies remain unknown. Although they are considered benign, the authors' review shows a 23% mortality rate. The authors report the successful use of carboplatinum and vincristine as treatment of this disease. PMID- 15543014 TI - Splenic rupture in a parental donor undergoing peripheral blood progenitor cell mobilization. AB - Peripheral blood progenitor cells (PBPCs) are now widely used as a source of progenitor cells for allogeneic transplantation. Recombinant human granulocyte colony-stimulating factor is used to mobilize PBPCs for collection by leukapheresis. Although side effects of mobilization are generally benign, adverse effects have been reported. The authors present a case of spontaneous splenic rupture, without splenomegaly, in a parental donor undergoing PBPC mobilization, review the literature regarding this adverse event, and explore issues regarding donor safety. PMID- 15543015 TI - Effect of fractionated ifosfamide on the pharmacokinetics of irinotecan in pediatric patients with osteosarcoma. AB - The combination of irinotecan (daily for 5 days for 2 consecutive weeks) and ifosfamide (daily on days 1 through 3) was investigated in children with osteosarcoma. Irinotecan pharmacokinetic investigations were performed before ifosfamide (day 1), after 3 days of ifosfamide (day 3), and 9 days after the end of ifosfamide (day 12). On day 3, the concentrations of irinotecan's active metabolite, SN-38, were below the limit of quantitation in two patients and were decreased in a third patient. The SN-38 area under the concentration-time curve remained below the day 1 value in two patients on day 12. The reduced area under the curve to the active metabolite SN-38 during ifosfamide therapy predicts a compromised efficacy of irinotecan in this combination. PMID- 15543016 TI - First report of a vincristine dose-related Raynaud's phenomenon in an adolescent with malignant brain tumor. AB - The authors report a case of a vincristine-induced, reproducible dose-related Raynaud's phenomenon. It occurred in a 14-year-old boy with a malignant brain tumor who received repeated vincristine injections. The authors describe how they handled this severe secondary Raynaud's phenomenon with acral cutaneous tissue necrosis. Reducing the dose of the vinca-alkaloid injections, together with an additional medication with a calcium-channel blocking agent, was a successful strategy in this patient. There are few case reports of secondary Raynaud's phenomenon in adult oncologic patients receiving certain anticancer drugs (including vincristine, bleomycin, and cisplatin), and to the authors' knowledge this kind of vincristine toxicity has not previously been described in either adults or children. PMID- 15543017 TI - Congenital peripheral primitive neuroectodermal tumor refractory to treatment. AB - The authors report an unusual case of an infant presenting with a congenital peripheral primitive neuroectodermal tumor (PPNET) of the left hand refractory to treatment. A newborn girl was born with a large bluish-red mass of 4.5 cm diameter protruding into the palm and the dorsum of the left hand. Tumor biopsy confirmed the diagnosis of PPNET. The initial metastatic workup for the detection of metastases was negative. Four cycles of chemotherapy according to CCSG 7881/POG 8850 regimen B were given. Despite this aggressive chemotherapy the tumor grew to involve the entire left hand. The left hand was amputated, and then two cycles of topotecan/cyclophosphamide were given. Five months later extensive metastases developed, involving the brain, lungs, liver, and skeleton, and the child died at the age of 2.5 years. PPNET presenting at birth is uncommon; presentation in the hand is unusual, and the fact that it did not respond to treatment is still more uncommon. PMID- 15543018 TI - A novel sickle hemoglobin: hemoglobin S-south end. AB - Sickle hemoglobin (Hb S; beta Glu6Val) is due to an AGTG; beta Lys132Asn, AAA>AAC). When present alone, the beta Lys132Asn mutation has low oxygen affinity. Therefore, this mutation may enhance the polymerization of the Hb S variant. Furthermore, the variant hemoglobin mimics Hb A on high-pressure liquid chromatography, and its identity is not easily diagnosed. A succinct review of variant sickle hemoglobins is also presented. PMID- 15543019 TI - Immune-mediated thrombocytopenia following dactinomycin therapy in a child with alveolar rhabdomyosarcoma: the unresolved issues. AB - The authors report a case of severe dactinomycin-induced thrombocytopenia in a child with alveolar rhabdomyosarcoma. The phenomenon is consistent with an immune process leading to the formation of platelet-specific antibodies. This study shows that this can be induced even with the first dose of actinomycin, and its persistence is unpredictably prolonged and does not correlate linearly in an inverted fashion with the platelet count. It will be important to identify the subsets of patients who can develop this phenomenon by molecular techniques and to define the exact mechanism in vitro leading to formation of these antibodies. This would facilitate profiling the therapy, preventing the need for multiple platelet transfusions with their obvious hazards. PMID- 15543020 TI - Severe lactic acidosis in a 14-year-old female with metastatic undifferentiated carcinoma of unknown primary. AB - A 14-year-old girl was found to have a large, non-tender breast mass with anemia and thrombocytopenia. The diagnosis of an undifferentiated carcinoma of unknown primary was made after open breast biopsy of the mass with negative immunohistochemical studies for breast malignancies. Further evaluation showed extensive metastatic disease affecting the bone marrow, ribs, liver, and brain with magnetic resonance imaging evidence of carcinomatous meningitis. Despite 2 months of chemotherapy and intensive supportive care, the patient died of severe lactic acidosis and disseminated intravascular coagulation after exaggerated menstrual bleeding. The association of severe lactic acidosis and undifferentiated carcinoma of unknown primary in an adolescent has not been previously described. PMID- 15543023 TI - Introduction to special issue on women's sexuality and outline of assessment of sexual problems. PMID- 15543024 TI - Recent advances in women's sexual function and dysfunction. AB - Current reconceptualization of women's sexual response acknowledges that women have many reasons or incentives for engaging in sex over and beyond sexual desire. Normative changes in their sexuality across the life span, with reproductive events, and with duration of relationship are recognized. Psychophysiological and preliminary functional magnetic resonance imaging data clarify that women's subjective experience of arousal may correlate poorly with signals reflective of genital congestion and also correlate poorly with activation of areas of the brain involved in organizing the reflexive genital vasocongestion. These aspects have been incorporated into new models of sexual response. Definitions of women's sexual dysfunction have recently been revised and expanded in keeping with these concepts. Mental well-being and other psychological and biological factors modulating desire, arousability, and response are areas of active research. Current understanding of the pathophysiology of chronic pain can be applied to the chronic intermittent pain and allodynia of chronic dyspareunia. PMID- 15543025 TI - Low sexual desire in midlife and older women: personality factors, psychosocial development, present sexuality. AB - OBJECTIVE: Recent population-based surveys indicate that the prevalence of sexual dysfunction, particularly low sexual desire and arousal disorders, is increasing with age. However, there seems to be greater variability of the sexual experience and functioning in midlife and older women, suggesting a higher dependence on basic conditions like general well-being, physical and mental health, quality of relationship, and life situation. DESIGN: A series of studies was conducted in the authors' Female Sexual Dysfunction research group to assess differences in (1) determinants of sexual satisfaction, (2) personality factors, and (3) present sexuality between younger and older women in both patient and nonpatient populations. RESULTS: The results of these studies highlight that in comparison with functional women, patients with hypoactive sexual desire are generally characterized by a vulnerable self-system with several rather inadequate self regulatory mechanisms. The results of the brief sexual function questionnaire indicate that the present sexuality of women seeking professional help for low sexual desire is significantly different from the sexuality of a control group of nonpatients. These between-group effects proved to be far more important than any age effects within both groups and showed that all domains of sexuality were negatively affected and overshadowed by the sexual dysfunction. CONCLUSIONS: These results are supportive of the growing evidence against a simple model of midlife sexuality that depicts women as victims of their bodily and hormonal changes. Instead, life stressors, contextual factors, past sexuality, and mental health problems are more significant predictors of midlife women's sexual interest than menopause status itself. Evaluation and treatment approaches require consideration of the full range of contextual factors, including relationship quality, personality factors, past experience, and mental and physical health. PMID- 15543026 TI - Genital vascular responsiveness and sexual feelings in midlife women: psychophysiologic, brain, and genital imaging studies. AB - The incidence of women's experiences of lack of sexual arousal increases in the years around the natural menopause transition. This raises the questions to what extent are sexual complaints in postmenopausal women related to hormonal changes, and what is the contribution of psychologic and contextual factors to sexual function in this transitional phase of life. This article reviews all published evidence regarding (1) the relation between menopausal status and sexual arousal symptoms, (2) the extent to which these sexual symptoms relate to measured genital vascular responses to sexual stimuli, and (3) the correlation between women's genital vascular response and their experience of subjective sexual arousal. Psychophysiologic and preliminary functional magnetic resonance imaging (MRI) studies of increases in genital congestion in response to erotic stimulation fail to identify differences between premenopausal and postmenopausal women. This suggests that although urogenital aging results in changes in anatomy and physiology of the genitals, postmenopausal women preserve their genital responsivity when sufficiently sexually stimulated. The vaginal dryness and dyspareunia experienced by some postmenopausal women may result from longstanding lack of sexual arousal and protection from pain previously afforded by estrogen related relatively high blood flow in the unaroused state. Psychophysiologic studies confirm similar increases in genital vasocongestion from erotic stimuli in women with and without chronically low sexual arousal, even in those women focusing specifically on their perceived lack of genital response. Moreover, studies repeatedly confirm highly variable correlations between subjective sexual arousal and measured increases in genital congestion. PMID- 15543027 TI - The effects of postmenopausal hormone therapies on female sexual functioning: a review of double-blind, randomized controlled trials. AB - Double-blind randomized controlled trials of estrogen and/or testosterone on sexual function among natural or surgical menopause in women are reviewed. Power, validity, hormone levels, and methodological issues were examined. Certain types of estrogen therapy were associated with increased frequency of sexual activity, enjoyment, desire, arousal, fantasies, satisfaction, vaginal lubrication, and feeling physically attractive, and reduced dyspareunia, vaginal dryness, and sexual problems. Certain types of testosterone therapy (combined with estrogen) were associated with higher frequency of sexual activity, satisfaction with that frequency of sexual activity, interest, enjoyment, desire, thoughts and fantasies, arousal, responsiveness, and pleasure. Whether specific serum hormone levels are related to sexual functioning and how these group effects apply to individual women are unclear. Other unknowns include long-term safety, optimal types, doses and routes of therapy, which women will be more likely to benefit from (or be put at risk), and the precise interplay between the two sex hormones. PMID- 15543028 TI - Sexual dysfunction in women with premature menopause. AB - OBJECTIVE: To discuss assessment and management of sexual dysfunctions associated with premature menopause. DESIGN: Literature review plus clinical observations. RESULTS: Studies on assessment and management of sexual dysfunctions associated with premature menopause are minimal. Premature menopause affects sexual identity, sexual function, and sexual relationship. Major modulating factors include etiology, life cycle stage, factors personal to the woman, and family and societal influences. Loss of ovarian function in adolescence may delay psychosexual maturity via psychological and biological mechanisms. Maternity becomes impossible, unless ovodonation is feasible, accepted, and legally available. Loss of sexual desire, subjective and genital arousal, and orgasm may be highly distressing. Reduction of sex hormones and psychological reactions of both partners to the prematurity and infertility are etiologically important. Estrogen therapy is usually indicated, but long-term safety data in this population are lacking. Testosterone therapy, also lacking in safety data and mainly investigational, may be needed to restore sexual arousability. Presence, age, and sexual health of the partner, having children, and quality of intimacy in previous and present relationships modulate coping attitudes and clinical outcome. CONCLUSIONS: An interdisciplinary medical and psychosexual approach comprises individualized hormone therapy and specific psychosexual treatment(s). Research into optimal management of sexual dysfunction with premature menopause is urgently needed. PMID- 15543029 TI - Women's sexual functioning, lifestyle, mid-age, and menopause in 12 European countries. AB - OBJECTIVE: To assess whether there are differences between women from different European countries in lifestyles and sexual functioning, and to identify the relative impact of social, hormonal, and lifestyle factors on mid-aged women's sexual functioning. DESIGN: Cross-sectional survey of a convenience sample of 601 women aged 45 to 60 years, consulting general practitioners in 12 European countries. Women completed the Short Personal Experiences Questionnaire and questions regarding social factors, lifestyle, attitudes, well-being, stress, health, and menopause status. RESULTS: Significant differences were found in psychosocial and lifestyle variables between countries. The only domain of sexual functioning that varied between countries was frequency of sexual intercourse (P < 0.05), the highest being for the group of Latin or southern countries (France, Portugal, Italy, Spain). General linear modeling found that change of partner, existence of partner, well-being, parity, and exercise were associated with both sexual response and frequency of sexual intercourse. Menopause status and stress were significantly associated with sexual response. Country of origin and body mass index were significantly associated with frequency of sexual intercourse. Structured equation modeling found that recalled experience on each sexual domain was the most important factor influencing that domain, whereas feelings for partner and change of partner had significant effects. A similar explanatory model was found for the sexual functioning for mid-aged European women as for Australian mid-aged women. CONCLUSIONS: There are significant differences between European countries in women's reports of lifestyles, psychosocial factors, and frequency of sexual intercourse but not in other aspects of their sexual response. These findings generate hypotheses to be tested in larger studies of female sexual functioning. PMID- 15543031 TI - Build it so they'll come. PMID- 15543033 TI - Hold a place of greatness for staff. AB - Lead your staff members to better team-work and their best performances. PMID- 15543034 TI - Anticipate responses to change. PMID- 15543035 TI - Take the workforce development challenge. PMID- 15543036 TI - Shared visions-new pathways illuminates the way to better hospitals. AB - Shared Visions-New Pathways focuses on continuous improvement rather than score achievement. PMID- 15543037 TI - Ethics paramount when patient lacks capacity. AB - Nurse leaders must use ethical decision-making skills when patients can't decide for themselves. PMID- 15543038 TI - Your hand in pain management. AB - Nurses in all practice settings encounter patient pain and struggle to manage it, despite medical advances. PMID- 15543040 TI - Congress 2004's silver anniversary a sparkling success. PMID- 15543041 TI - Can't we all just get along? Know your role in fostering hospital-provider collaboration. AB - When nurses and physicians, both staff and management, work together toward mutually acceptable and productive goals, the patient and the institution prosper. PMID- 15543042 TI - Restraint regulation: the tie that binds. AB - Restraint litigation ranks among nurses' most common legal concerns. Better understand legal and regulatory standards to safely incorporate them into policies, procedures, practice, and education. PMID- 15543044 TI - Break down the barriers to CPR compliance. AB - Review the steps nurse leaders at one facility implemented to greatly improve the organization's CPR compliance. PMID- 15543046 TI - Using technology to help alleviate pain. AB - Learn the latest pain management techniques of PCA pumps and implanted systems. PMID- 15543048 TI - Staff in-services: the basics for betterment. AB - Learn how to turn your experiences at a nursing conference into a worthy in service for your staff members. PMID- 15543052 TI - Flexion-extension response of the thoracolumbar spine under compressive follower preload. AB - STUDY DESIGN: The authors conducted an in vitro biomechanical flexibility study of T2-S1 specimens in flexion-extension under compressive follower preloads of physiological magnitudes. OBJECTIVES: The objectives of this study were to test the hypotheses that 1) the thoracolumbar spine will support compressive preloads of in vivo magnitudes and 2) allow physiological mobility under flexion-extension moments if the preload is applied along an optimized follower load path that approximates the kypholordotic curve of the thoracolumbar spine. SUMMARY OF BACKGROUND DATA: In the absence of muscle forces, the ligamentous thoracolumbar spine specimens cannot support the compressive loads expected in vivo. As a result, the flexibility of the thoracolumbar spine in flexion-extension has not been studied in vitro under physiological compressive preloads. METHODS: Seven human thoracolumbar spines (T2-sacrum) were subjected to flexion and extension moments (up to 8 and 6 Nm, respectively) under compressive preloads from 0 to 800 N applied along an optimized follower preload path. The experimental technique applied the compressive preload such that: 1) it minimized the internal shear forces and bending moments resulting from the preload application, 2) made the internal force resultant compressive, and 3) caused the preload path to approximate the tangent to the curve of the thoracolumbar spine. The range of motion was measured in the T2-sacrum, T2-T11, T11-L1, and L1-sacrum regions. RESULTS: All thoracolumbar specimens supported the compressive follower preload up to 800 N without damage or instability. At 800 N preload, the total flexion extension range of motion of the T2-sacrum region decreased by 22%, from a mean of 73 degrees to 57 degrees (P < 0.05). The range of motion of the T2-T11 and L1 sacrum regions decreased from the baseline value by 23% and 30%, respectively, at a preload of 800 N. The sagittal mobility of the thoracolumbar junction (T11-L1) was not affected by the preload. The follower preload did not significantly affect the proportion of the total T2-sacrum flexion-extension range of motion contributed by the T2-T11 and L1-sacrum regions of the thoracolumbar spine. CONCLUSIONS: The optimized follower preload vector minimizes the effects of artifact moment and shear force on the range of motion of the thoracolumbar spine in flexion-extension. This model allows the entire thoracolumbar spine to be investigated under physiological loading for different clinical applications. PMID- 15543053 TI - Changes in the cross-sectional area of multifidus and psoas in patients with unilateral back pain: the relationship to pain and disability. AB - STUDY DESIGN: Prospective, cross-sectional observational study. OBJECTIVES: The aim of this study was to determine if there was an association between wasting of psoas and multifidus as observed on MRI scans and the presenting symptoms, reported pathology, pain, or disability of a cohort of patients presenting with unilateral low back pain. SUMMARY OF BACKGROUND DATA: Current physiotherapy practice is often based on localized spine stabilizing muscle exercises; most attention has been focused on transversus abdominus and multifidus with relatively little on psoas. METHOD: Fifty consecutive patients presenting to a back pain triage clinic with unilateral low back pain lasting more than 12 weeks were recruited. The cross-sectional surface area (CSA) of the muscles was measured. Duration of symptoms, rating of pain, self-reported function, and the presence of neural compression were recorded. RESULTS: Data analysis compared the CSA between the symptomatic and asymptomatic sides. There was a statistically significant difference in CSA between the sides (P < 0.001). There was a positive correlation between the percentage decrease in CSA of psoas on the affected side and with the rating of pain (rho = 0.608, P < 0.01), reported nerve root compression (rho = 0.812, P < 0.01), and the duration of symptoms (rho = 0.886, P < 0.01). There was an association between decrease in the CSA of multifidus and duration of symptoms. CONCLUSIONS: Atrophy of multifidus has been used as one of the rationales for spine stabilization exercises. The evidence of coexisting atrophy of psoas and multifidus suggests that a future area for study should be selective exercise training of psoas, which is less commonly used in clinical practice. PMID- 15543054 TI - Mobile and reducible atlantoaxial dislocation in presence of occipitalized atlas: report on treatment of eight cases by direct lateral mass plate and screw fixation. AB - STUDY DESIGN: This is a retrospective review on the application of the authors' technique of atlantoaxial lateral mass plate and screw fixation on a rare subset of patients having mobile and reducible atlantoaxial subluxation in the presence of occipitalized atlas. OBJECTIVE: Mobile and reducible atlantoaxial dislocation in the presence of an occipitalized atlas is extremely rare. We retrospectively analyzed the management of eight such patients who were treated by lateral mass plate and screw fixation. SUMMARY OF BACKGROUND DATA: The authors report their experience with a rare congenital craniovertebral anomaly where there was a mobile and reducible atlantoaxial dislocation in the presence of the occipitalized atlas. The cases were treated by lateral mass plate and screw method of treatment described by the authors in 1994. METHODS: Eight patients with occipitalized atlas had a mobile and completely reducible atlantoaxial dislocation. These patients were treated between the years 1996 to 2003 and underwent a direct atlantoaxial lateral mass fixation. The follow-up ranges from 4 months to 6 years (average, 48 months). RESULTS: All patients underwent a successful fixation and ultimately achieved a solid bony fusion. There were no intraoperative or postoperative complications. CONCLUSIONS: Although lateral mass plate and screw fixation in the presence of occipitalized atlas is technically a relatively difficult and anatomically a precise surgical procedure, the firm and segmental stabilization that it provides offers an optimum situation for bony fusion. PMID- 15543055 TI - Juxtaposition of an epidural intraforaminal cavernous hemangioma and an intradural schwannoma. AB - STUDY DESIGN: A case of juxtaposition of a purely extradural cavernous hemangioma and an intradural schwannoma in the thoracic spine is reported. OBJECTIVES: The objectives of this study were to present a rare and before-surgery unexpected combination of spinal tumors, which may complicate surgical removal; and to discuss the radiologic features and the possible pathogenesis of this combination of tumors and its implications on surgical therapy. SUMMARY OF BACKGROUND DATA: Juxtaposition of different spinal tumors is exceedingly rare in patients without neurofibromatosis. To the authors' knowledge, no combination of a purely epidural spinal cavernous hemangioma with an intradural schwannoma has been previously reported. MATERIAL AND METHODS: A 47-year-old woman presented with progressive paraparesis. Magnetic resonance imaging revealed a partly cystic intradural mass at the T6-T7 level and a solid extradural intraforaminal component that demonstrated slightly different signal characteristics. An intra-/extradural schwannoma was assumed. RESULTS: Surgical exposure displayed a highly vascularized lobulated mass within the left neuroforamen at T6-T7. There was no continuity with the larger intradural, cystic, and mildly vascularized lesion. Histologic examination diagnosed an extradural cavernous hemangioma and an intradural schwannoma. CONCLUSIONS: The segmental juxtaposition of lesions should be considered in the differential diagnosis of spinal pathology. PMID- 15543056 TI - Traumatic epidural emphysema. AB - STUDY DESIGN: This is a case report. OBJECTIVE: A rare case of epidural emphysema secondary to traumatic pneumomediastinum in the absence of pneumothorax is reported. SUMMARY OF BACKGROUND DATA: Epidural air secondary to traumatic pneumomediastinum in the absence of pneumothorax has only been reported previously by Willing. Other causes of traumatic epidural emphysema include pneumothorax, pelvic fracture, dural enteric fistula, and herniation of a spinal disc. METHODS: A young man who sustained blunt cervical and chest trauma after a basketball game accident was found to have pneumomediastinum and surgical emphysema on a chest radiograph and underwent computed tomography (CT). RESULTS: The CT scan demonstrated surgical emphysema, pneumomediastinum, and epidural emphysema with no pneumothorax. The patient was managed conservatively, and the epidural emphysema and pneumomediastinum resolved spontaneously. CONCLUSION: Epidural emphysema secondary to traumatic pneumomediastinum is benign and self limiting. However, the life-threatening causes should be considered and ruled out. PMID- 15543058 TI - Selective inhibition of tumor necrosis factor-alpha prevents nucleus pulposus induced histologic changes in the dorsal root ganglion. AB - STUDY DESIGN: The possibility to prevent nucleus pulposus-induced structural changes of the dorsal root ganglion (DRG) by selective tumor necrosis factor alpha (TNF-alpha) inhibition was assessed in an experimental model in the rat spine. OBJECTIVES: To evaluate the role of TNF-alpha in the mediation of nucleus pulposus-induced structural changes by using selective inhibition and to confirm the effect of TNF-alpha inhibitor at the point of histologic findings. SUMMARY OF BACKGROUND DATA: TNF-alpha is known to be released from the nucleus pulposus, and has been suggested to play a key role in chemical damage of the adjacent nerve tissue. The TNF-alpha inhibitor prevents the reduction of nerve conduction velocity and may limit the nerve fiber injury, intracapillary thrombus formation, and intraneural edema formation caused by nucleus pulposus. However, there is no report on the effect of the inhibitor regarding histologic findings and the appearance of the TNF-alpha in the DRG exposed to nucleus pulposus. METHODS: 1) Rats were treated with an intraperitoneal injection of infliximab. Nucleus pulposus from the disc was obtained 1, 3, 7, 14, and 21 days after the injection. The TNF-alpha-positive cells were observed using immunohistochemistry. 2) Disc herniation of the nucleus pulposus was made on the L4-L5 disc in rats. Two groups were treated with selective TNF-alpha inhibitor 1 day before or 3 hours after surgery. The other group received no TNF-alpha inhibitor. The L4 DRG was resected 1, 3, 7, 14, and 21 days after surgery. The specimens were processed for hematoxylin and eosin staining and immunohistochemistry using rabbit antisera to TNF-alpha. The histologic findings and TNF-alpha-positive cells were observed by light microscopy. RESULTS: 1) While positively stained immunoreactive TNF-alpha appeared between 7 and 21 days, no immunoreactive TNF-alpha was observed 1 and 3 days after injection in the nucleus pulposus. 2) The histologic changes of the DRG caused by nucleus pulposus were smaller in the infliximab treatment group than those in the nontreatment group. The number of immunoreactive TNF-alpha cells was high 1 and 3 days after surgery in the DRGs of disc herniation rats that were treated without an injection of the inhibitor, low on day 7 and 14, and very low on day 21 after surgery. No immunoreactive TNF-alpha was observed in the DRGs of the TNF-alpha inhibitor treatment groups on day 1, 3, and 21 after surgery. Weakly stained cells were sometimes observed in rats at day 7 and 14 after surgery. CONCLUSIONS: Infliximab may prevent the histologic damage induced by nucleus pulposus. When rats were given a single intraperitoneal injection of infliximab at the beginning of disc herniation, the histologic damage seemed to be reduced in comparison with the nontreated rats. PMID- 15543059 TI - Modeling of the sagittal cervical spine as a method to discriminate hypolordosis: results of elliptical and circular modeling in 72 asymptomatic subjects, 52 acute neck pain subjects, and 70 chronic neck pain subjects. AB - STUDY DESIGN: Computer analysis of digitized vertebral body corners on lateral cervical radiographs. OBJECTIVES: Using elliptical and circular modeling, the geometric shape of the path of the posterior bodies of C2-C7 was sought in normal, acute pain, and chronic pain subjects. To determine the least squares error per point for paths of geometric shapes, minor axis to major axis elliptical ratios (b/a), Cobb angles, sagittal balance of C2 above C7, and posterior tangent segmental and global angles. SUMMARY OF BACKGROUND DATA: When restricted to cervical lordotic configurations, normal, acute pain, and chronic pain subjects have not been compared for similarities or differences of these parameters. Conventional Cobb angles provide only a comparison of the endplates of the distal vertebrae, while geometric modeling provides the shape of the entire sagittal curves, the orientation of the spine, and segmental angles. METHODS: Radiographs of 72 normal subjects, 52 acute neck pain subjects, and 70 chronic neck pain subjects were digitized. For normal subjects, the inclusion criteria were no kyphotic cervical segments, no cranial-cervical symptoms, and less than +/- 10 mm horizontal displacement of C2 above C7. In pain subjects, inclusion criteria were no kyphotic cervical segments and less than 25 mm of horizontal displacement of C2 above C7. Measurements included segmental angles, global angles of lordosis (C1-C7 and C2-C7), height-to-length ratios, anterior weight bearing, and from modeling, circular center, and radius of curvature. RESULTS: In the normal group, a family of ellipses was found to closely approximate the posterior body margins of C2-C7 with a least squares error of less than 1 mm per vertebral body point. The only ellipse/circle found to include T1, with a least squares error of less than 1 mm, was a circle. Compared with the normal group, the pain group's mean radiographic angles were reduced and the radius of curvature was larger. For normal, acute, and chronic pain groups, the mean angles between posterior tangents on C2-C7 were 34.5 degrees, 28.6 degrees, and 22.0 degrees, C2-C7 Cobb angles were 26.8 degrees, 16.5 degrees, and 12.7 degrees, and radius of curvature were r = 132.8 mm, r = 179 mm, and r = 245.4 mm, respectively. CONCLUSIONS: The mean cervical lordosis for all groups could be closely modeled with a circle. Pain groups had hypolordosis and larger radiuses of curvature compared with the normal group. Circular modeling may be a valuable tool in the discrimination between normal lordosis and hypolordosis in normal and pain subjects. PMID- 15543060 TI - Results of surgical treatment for degenerative cervical myelopathy: anterior cervical corpectomy and stabilization. AB - STUDY DESIGN: This retrospective study involves 26 patients with degenerative cervical myelopathy who were surgically treated by anterior corpectomy, titanium mesh cage (TMC) filled with autogenous bone, and anterior plate +/- posterolateral plate and fusion. OBJECTIVES: This study was conducted to determine the indications, efficacy, and complication rate associated with performing corpectomy to achieve anterior decompression of neural elements or for removing anterior lesions. SUMMARY OF BACKGROUND DATA: This retrospective study involves patients with degenerative cervical myelopathy who were surgically treated by > or =2-level anterior corpectomy, TMC filled with autogenous bone, and anterior plate +/- posterolateral plate and fusion. The purpose was to evaluate and compare the results in terms of neurologic recovery and function and effectivity of TMC as a structural support. METHODS: Twenty-six patients with degenerative cervical myelopathy who had surgical treatment and average 30 months (range, 24-52 months) follow up were included. The mean age was 64.9 years (range, 55-74 years) and average period between myelopathic symptoms and surgery was 2.8 years (range, 6 months-5 years). Preoperative evaluation of every patient consisted of anterior-posterior, lateral, bilateral oblique, flexion, and extension radiographs, computed tomography reconstructions and magnetic resonance imaging of the cervical spine, Doppler ultrasound of the carotid arteries, vertebral artery magnetic resonance angiography, neurologic examination, and electromyography. Degree of pre- and postoperative myelopathy was determined according to the scoring systems developed by Nurick and Japanese Orthopedic Association (JOA). Twelve patients had a mild balance problem and difficulty while walking but were able to perform their daily activities. Fourteen patients had spastic quadriparesis ambulating on either crutches or with wheelchairs. Of these, 11 experienced bladder disturbance as well. Surgical treatment in 18 patients consisted of anterior decompressive corpectomy, structural TMC, and anterior plate stabilization in 14 patients who had 2-level corpectomy. Posterior plate stabilization without laminectomy was added to this procedure in another 4 patients who had 3- or more level corpectomy. The remaining 8 patients had first laminectomy and posterolateral plate, then anterior corpectomy, TMC, and anterior plate on the same stage. Corpectomy levels were between C3 and T1, and anterior corpectomy, structural TMC, and anterior plating was the procedure that all patients had in common. RESULTS: Mean sagittal Cobb angle (C2-C7) was 9 degrees (range, 0-23 degrees) before surgery, 17.1 degrees (range, 11-22 degrees) on the third postoperative month, and 16.9 degrees (range, 10-22 degrees) at last follow up. The difference in sagittal alignment on the third month and last follow up was not statistically significant (P > 0.05). Average preoperative Nurick score was 3.5 (range, 2-5) and JOA score was 7 (range, 1-14). Major and statistically significant neurologic recovery was within the first 3 months, and average Nurick and JOA scores at 3 months were 2 (range, 0-3) and 11 (range, 8-17) (P < 0.001), respectively. All patients had improved neurologic status at final follow up. As confirmed by plain radiographs and computed tomography reconstructions, solid fusion was achieved across the TMC with no settling or migration, and we had no implant-related complication or failure. As major complications, 1 (3.8%) early deep posterior infection developed but responded to early debridement and antibiotics. Also, 3 patients (11.5%) had transient C5 nerve root injury. At final follow up, all patients were able to ambulate without support and maintain their daily activities. CONCLUSIONS: Anterior decompression provides good neurologic recovery in patients with degenerative cervical myelopathy. TMC provides good structural support, and solid fusion can be achieved with TMC and anterior plate (for < or =2-level corpectomy) and/or posterior plate (> or =3 level corpectomy). There is increased risk of C5 nerve root injury when first laminectomy and posterolateral plate stabilization are performed. PMID- 15543061 TI - Integrated outcome assessment after anterior cervical discectomy and fusion: myelocompression but not adjacent instability affect patient-reported quality of life and cervical spine symptoms. AB - STUDY DESIGN: The authors conducted a cross-sectional study. OBJECTIVE: Integrated assessment of adjacent instability (AI), myelocompression (MC), magnetic resonance imaging (MRI) signs of myelopathy (MRISM), physician-assessed clinical signs and symptoms, including clinical signs of myelopathy (CSM), patients' self-reported symptoms and quality of life after anterior cervical discectomy and fusion (ACDF). MATERIALS AND METHODS: Fifty-four patients who had ACDF between 1986 and 1995 received MRI scans, conventional and flexion/extension radiographs to assess myelocompression, MRISM, fusion, and AI. Clinical outcome was assessed using signs and symptoms based on selected items of Odom's criteria, Oswestry low back pain disability questionnaire, and the neck disability index. Patients reported their quality of life (QL) on a standardized instrument (Profiles of QL of Chronically Ill [PLC]) and by a specific validated Cervical Spine Symptom Scale (CSSS). RESULTS: Myelocompression was found more frequently than expected (24%). MRISM were seen in 2 patients (4%). One of the 2 patients developed CSM. Fusion was achieved in 94% (with kyphosis in 17%). AI was found in 30%. However, only myelocompression but not AI was associated with statistically significant decreases in most QL scores (i.e., everyday capabilities, positive mood) and high cervical spine symptom burden (all P's < 0.05). CONCLUSIONS: The study results underline the need for a better understanding of the biomechanical changes in the adjacent unfused segments. Consensus is needed on postoperative follow-up guidelines, including pain management strategies. Future studies on the outcome of ACDF will profit from an integrated outcome approach, including assessments based on imaging, physicians, and patients. PMID- 15543062 TI - Transpedicular screw placement evaluated by axial computed tomography of the cervical pedicle. AB - STUDY DESIGN: We evaluated the trajectory of transpedicular screws in the cervical spine using axial computed tomography (CT). OBJECTIVES: To provide a safe transpedicular screw trajectory by measuring the dimensions of the cervical pedicle and evaluating the entrance points and the insertion angles of transpedicular screws. SUMMARY OF BACKGROUND DATA: The morphology of the cervical pedicle has been studied, but few in vivo CT-based studies of pedicle dimensions and transpedicular screw placement in the cervical spine have been reported. METHODS: The dimensions of the pedicles (C3-C7) were determined in 30 patients with cervical spinal lesions from CT images. The space available for transpedicular screws (SAS) was defined as the distance between two parallel lines tangential to the spinal canal and the transverse foramen, respectively. SAS was evaluated at 25 degrees and 50 degrees insertion angles. RESULTS: SAS at a 25 degrees insertion angle (SAS-25) ranged from 4.7 to 5.4 mm. SAS at 50 degrees (SAS-50) ranged from 6.1 to 6.6 mm. SAS-25 and SAS-50 were significantly different. Four-millimeter-diameter screws would fit in all 120 C3-C6 vertebrae studied at 50 degrees, but 20 (17%) would not fit at 25 degrees. CONCLUSIONS: Axial CT measurements should facilitate transpedicular screw fixation in the cervical spine. We believe that the screw insertion angle should be close to 50 degrees, which is the mean pedicle transverse angle from C3-C6. The entry point of the pedicle screw should be located as laterally as possible in the posterior surface of the lateral mass. PMID- 15543064 TI - Reoperation rate after instrumented posterior lumbar interbody fusion: a report on 1680 cases. AB - STUDY DESIGN: Retrospective study on the incidence of reoperation in patients previously treated by instrumented posterior lumbar interbody fusion. OBJECTIVES: To answer the following questions: Reoperation rate after PLIF? And is there any influence of the length of fusion on the reoperation rate? SUMMARY OF BACKGROUND DATA: The use of different techniques demonstrates that there is currently no ideal procedure for lumbar fusion. The instrumented posterior lumbar interbody fusion (PLIF) shows a comparable success rate to the so-called 360 degrees fusion techniques (combined dorsoventral spondylodesis) without the need of an anterior approach. METHODS: We reviewed 1680 patients who underwent a PLIF at our institution between January 1995 and December 2000. A total of 3053 levels were fused. The reoperation rate was analyzed. The mean follow-up was 5 years. RESULTS: There were 221 (13.2%) reoperations in 206 patients (12.2%). Of 1680 PLIFs, 312 were multisegmental (>2 segments). Within this group, 45 (14.4%) revisions were done. We found that the most important difference between the multisegmental PLIFs and the mono- or bisegmental PLIFs is the rate of adjacent segment decompensation (5.1% vs. 2.3%), and this was statistically significant. The reoperation rate between those two groups was only slightly different with 12.9% for mono- or bisegmental and 14.4% for multisegmental PLIFs. CONCLUSIONS: The fusion length does not show a significant difference in the reoperation rate as such. Nevertheless, we registered a significantly higher incidence for decompensation of adjacent segments after multisegmental PLIFs. PMID- 15543065 TI - Correlation of the amount of disc removed in a lumbar microdiscectomy with long term outcome. AB - STUDY DESIGN: The authors conducted a prospective clinical study. OBJECTIVES: The objectives of this study were to investigate the relationship, if any, of the amount of removed disc in a standard first-time lumbar microdiscectomy and long term outcome as well as recurrence and postoperative instability rates. SUMMARY OF BACKGROUND DATA: There is lack of data on the amount of disc that needs to be removed during a lumbar microdiscectomy. Anecdotal data and dogmatic recommendations make the subject even more controversial. MATERIAL AND METHODS: A total of 106 patients underwent a standard first-time lumbar microdiscectomy for medically refractory sciatica resulting from a herniated disc. The removed disc material was weighted. All patients were clinically followed for at least 2 years and outcome was evaluated by using pain intensity, presence of physical signs, functional capacity, return to work, and patients' opinion regarding their outcome. RESULTS: The mean amount of disc removed was calculated at 2.1 +/- 0.9 g. Ninety-one patients had an excellent outcome and returned to their preoperative work. Fifteen patients had persistent symptomatology and underwent extensive radiographic workup, which revealed a disc recurrence at the same level in 8 patients and first-degree instability in 3 patients. In the remaining 4 patients, no clinical or radiographic abnormality was proven and the patients were treated conservatively, whereas all of them had applied for disability. No relationship was proven between the amount of the removed disc and the intraoperative blood loss or the intraoperative complication rate. Our statistical analysis showed no correlation between the amount of the removed disc and the long- term outcome, recurrence rate, or postoperative instability. CONCLUSIONS: The degree of disc removal did not influence the outcome or complication rate in our clinical series. PMID- 15543067 TI - Relation between laminectomy and development of adjacent segment instability after lumbar fusion with pedicle fixation. AB - STUDY DESIGN: A retrospective study of 101 patients who had undergone posterolateral lumbar fusion, to analyze the association between adjacent instability and the extent of laminectomy. OBJECTIVES: To investigate the hypothesis that the integrity of the posterior complex (spinous process/supraspinous ligament/spinous process) between the fused segments and the neighboring motion segments significantly influences lumbar spine stability. SUMMARY OF BACKGROUND DATA: Spinal fusion with pedicle fixation accelerates the degeneration of adjacent motion segments. The lowest cranial motion segment is the most common level for the development of adjacent instability. Laminectomy, including removal of the spinous process, supraspinous ligament, interspinous ligament, lamina and ligamentum flavum, jeopardizes the integrity of the posterior complex of the spine. METHODS: This study enrolled 101 patients, followed up for at least 6 years, who had been treated with posterolateral lumbar fusion with pedicle fixation because of lumbar spondylolisthesis. The diagnosis of adjacent instability depended on the dynamic lateral views of the lumbosacral spine during each follow-up. The integrity of the posterior complex was based on the extent of laminectomy and the fusion level. RESULTS: At the cranial adjacent motion segment, 2 of 31 (6.5%) patients with preserved posterior complex integrity between the fused segment and motion segment developed adjacent instability, compared with 17 of 70 (24.3%) without preserved posterior complex integrity. At the caudal adjacent motion segment, none of 13 patients (0%) with preserved integrity developed adjacent instability, compared with 3 of 54 (5.6%) without preserved integrity. CONCLUSIONS: Damaging the integrity of the posterior complex between the fused segments and the neighboring motion segments may jeopardize lumbar spine stability. Sacrificing either the supraspinous ligament or the tendon insertion points on the spinous processes leads to an accelerated development of adjacent instability. PMID- 15543069 TI - Bacterial spondylodiscitis in the patients with hemodialysis. AB - STUDY DESIGN: Retrospective clinical and laboratory data analyses were performed on hemodialysis (HD) patients with bacterial spondylodiscitis. OBJECTIVES: The purpose of this study was to investigate clinical characteristics and related problems for the diagnosis and treatment of spondylodiscitis in patients on maintenance HD. Possible factors for the development of spondylodiscitis were also discussed. SUMMARY OF BACKGROUND DATA: Although bacterial spondylodiscitis is one of most serious complications that can occur in HD patients, few reports are seen describing its clinical course and treatment in HD patients. METHODS: A total of 9 HD patients were diagnosed as having bacterial spondylodiscitis at our institute. The onset of infection, characteristics of clinical symptoms, and clinical course were reviewed retrospectively. RESULTS: Latent form occurrence was most frequent, and only 1 in 9 cases presented high-grade fever at the beginning of treatment. Many complications were encountered both in conservatively treated and operated cases. Three patients were operated on, 1 of whom died 2 days after operation. Two of six patients in the conservatively treated group also died during the treatment period. CONCLUSIONS: The presence of bacterial spondylodiscitis must be considered when treating back pain of HD patients even when they are afebrile. Careful observation of general status in addition to local conditions is essential. Indication of operation should be considered carefully because of the poor general status and bone quality of HD patients. MRI, in addition to plain radiographs, was necessary to differentiate destructive spondylarthropathy from bacterial spondylodiscitis. PMID- 15543070 TI - The prevalence of contraindications to total disc replacement in a cohort of lumbar surgical patients. AB - STUDY DESIGN: This is a retrospective review of the epidemiology of contraindications to lumbar total disc replacement (TDR). OBJECTIVE: To define the prevalence of contraindications to lumbar total disc replacement in a cohort of patients undergoing lumbar surgery in the senior author's (F.P.C.) practice. SUMMARY OF BACKGROUND DATA: No published reports have documented the prevalence of contraindications to lumbar total disc replacement. METHODS: We performed a retrospective review of 100 consecutive patients who had lumbar surgery by one surgeon between September and December 2002. Procedures performed and contraindications to TDR were recorded. Contraindications to TDR included central or lateral recess stenosis, facet arthrosis, spondylolysis or spondylolisthesis, herniated nucleus pulposus with radiculopathy, scoliosis, osteoporosis, and postsurgical pseudarthrosis or deficiency of posterior elements. Patients were divided into fusion and nonfusion groups. The percentage of patients without contraindications to TDR was calculated. RESULTS: Of 100 patients, 56 had fusions and 44 had nonfusion surgery. In the fusion group, 56 of 56 patients had contraindications to TDR. In the nonfusion group, 11% (5 of 44) were candidates for TDR. Overall, 5% of patients in this series were candidates for TDR. The average number of contraindications to TDR was 2.48 (range, 0-5). CONCLUSIONS: Predictions that TDR will replace fusion are premature. A small percentage (5%) of the patients currently indicated for lumbar surgery at our institution have no contraindications to TDR. Future growth in TDR implantation will result from the indication of patients for surgery who would not be indicated today or from the elimination of current contraindications. PMID- 15543071 TI - Complications associated with lumbar laminectomy: a comparison of spinal versus general anesthesia. AB - STUDY DESIGN: A case-controlled, comparative study of 400 patients undergoing lumbar surgery, treated with either spinal or general anesthesia. An independent observer analyzed outcomes. OBJECTIVES: To determine the rate and type, of perioperative complications associated with each anesthetic method among lumbar surgery patients. SUMMARY OF BACKGROUND DATA: Spinal anesthesia is infrequently used for spinal procedures. While complications associated with spinal anesthesia are rare, some authors have suggested that spinal anesthesia may exacerbate existing neurologic disease and have recommended against its use in lumbar disc surgery. Others have found the technique safe and effective. General anesthesia may be preferred because it is seen as the routine accepted practice, because of greater patient acceptance and the ability to perform longer operations, or because of a general sense that general anesthesia is "safer" in these procedures. METHODS: Patients treated between 1994 and 1998 were matched for anesthetic class, preoperative diagnosis, surgical procedure, and perioperative protocols. All patients were treated according to a uniform protocol and recovered in the same perianesthetic environment. Data from the intraoperative period through hospital discharge were collected and compared. RESULTS: A total of 200 patients were included in each group. Overall complication rates and time to discharge were significantly lower in spinal anesthetic patients. Total anesthetic and operative times were significantly longer for general anesthetic patients, and perioperative heart rate and mean arterial pressures were elevated compared with those in spinal anesthetic patients. Nausea, requirements for antiemetic medication, and the incidence of urinary retention were significantly increased among general anesthesia patients. Spinal anesthesia patients had fewer spinal headaches compared with the general anesthetic group, but statistical significance was not obtained. CONCLUSIONS: For patients undergoing decompressive lumbar surgery, spinal anesthesia is at least comparable to general anesthetic with respect to complications. Specific advantages to spinal anesthesia include decreased nausea and antiemetic requirements, reduced analgesic requirements, and reduced overall complication rate. PMID- 15543072 TI - The assessment of intraobserver and interobserver error in the measurement of noncongenital scoliosis in children < or = 10 years of age. AB - STUDY DESIGN: Retrospective review of scoliosis radiographs. OBJECTIVES: To determine measurement variability in children < or = 10 years of age with noncongenital scoliosis. SUMMARY OF BACKGROUND DATA: Measurement variability in congenital and adolescent idiopathic scoliosis has been studied. There is no study of measurement variability in young children with noncongenital scoliosis. METHODS: A retrospective review of children < or = 10 years of age followed for noncongenital scoliosis was performed. End vertebrae were identified on radiographs, and the curves were measured (Cobb method) twice by each of six observers. The same soft lead pencil and goniometer was used. Intraobserver and interobserver variability for continuous data was determined. RESULTS: There were 64 children. The diagnosis was infantile/juvenile idiopathic scoliosis in 42, neuromuscular scoliosis in 7, scoliosis associated with mesenchymal disorders or other syndromes in 12, and unknown in 3 children. The curve was thoracic in 54, thoracolumbar in 8, and lumbar in 2. There were 19 left and 45 right curves. The average age was 6.6 +/- 2.6 years. There were a total of 768 Cobb angle measurements with an average Cobb angle of 38 +/- 22 degrees (range, 10 degrees 115 degrees ). Intraobserver variability was +/- 6 degrees; interobserver variability was +/- 7 degrees. CONCLUSION: In children < or =10 years of age with noncongenital scoliosis, intraobserver measurement variability in Cobb angle measurement is +/- 6 degrees and interobserver variability is +/-7 degrees. To be certain that there is a significant difference between Cobb angle measurements in children with noncongenital scoliosis and < or = 10 years of age there must be a change of at least +/-7 degrees. PMID- 15543073 TI - Radiographic changes in the lumbar spine in former elite athletes. AB - STUDY DESIGN: The authors conducted a retrospective cohort study. OBJECTIVE: The objective of this study was to clarify the occurrence of radiographic changes (vertebral osteophytes, heights of lumbar discs, concavity index) of the lumbar spine in former elite athletes of different track and field disciplines. SUMMARY OF BACKGROUND DATA: The influence of physical activity on occurrence of radiographic changes in the lumbar spine is not well known and seems to be contradictory. The loadings in the different track and field disciplines seem to play an important role in the development of radiographic changes. METHODS: One hundred fifty-nine former male elite track and field athletes were selected for a radiologic study. The heights of lumbar discs, the concavity index, the presence of anterior vertebral osteophytes, a radiographic evaluation according to Kellgren and Lawrence, and the FFbH-R score for the assessment of functional limitations in activities of daily living (ADLs) were determined. The influence of age, body mass index, current physical activity, and training history was also examined. RESULTS: In high jumpers and throwers, the absolute heights of lumbar discs increased from level T12/L1 to a maximum at L4/L5 and decreased again from level L4/L5-L5/S1. In endurance athletes and other jumpers, the absolute heights increased linearly from level T12/L1 to a maximum at L5/S1. The concavity index did not yield any significant differences between athlete categories. Shot putters, discus throwers, and high jumpers showed a significantly higher prevalence of osteophytes after adjustment for possible confounders. According to Kellgren and Lawrence, the highest prevalence of radiographic changes in the lumbar spine is seen in javelin throwers. Significant differences in the assessment of functional limitations in ADLs are not found between the disciplines. CONCLUSION: In throwing disciplines, the lumbar spine is more highly loaded than in jumpers and runners. Despite the observation of evident degenerative changes in some former athletes, there were only minor changes seen in ADLs. Even if body constitution is taken as a preselection factor, athletes in throwing disciplines as well as high jumpers have a higher risk of developing vertebral osteophytes of the lumbar spine. PMID- 15543074 TI - Changes in recruitment of the abdominal muscles in people with low back pain: ultrasound measurement of muscle activity. AB - STUDY DESIGN: Ultrasound and electromyographic (EMG) measures of trunk muscle activity were compared between low back pain (LBP) and control subjects in a cross-sectional study. OBJECTIVES: To compare the recruitment of the abdominal muscles (measured as a change in thickness with ultrasound imaging) between people with and without low back pain and to compare these measurements with EMG recordings made with intramuscular electrodes. SUMMARY OF BACKGROUND DATA: Although ultrasonography has been advocated as a noninvasive measure of abdominal muscle activity, it is not known whether it can provide a valid measure of changes in motor control of the abdominal muscles in LBP. METHODS: Ten subjects with recurrent LBP and 10 matched controls were tested during isometric low load tasks with their limbs suspended. Changes in thickness from resting baseline values were obtained for transversus abdominis (TrA), obliquus internus (OI), and obliquus externus (OE) using ultrasonography. Fine wire EMG was measured concurrently. RESULTS: Study participants with LBP had a significantly smaller increase in TrA thickness with isometric leg tasks compared with controls. No difference was found between groups for OI or OE. Similar results were found for EMG. People with LBP had less TrA EMG activity with leg tasks, and there was no difference between groups for EMG activity for OI or OE. CONCLUSIONS: This study reinforces evidence for changes in automatic control of TrA in people with LBP. Furthermore, the data establish a new test of recruitment of the abdominal muscles in people with LBP. This test presents a feasible noninvasive test of automatic recruitment of the abdominal muscles. PMID- 15543075 TI - Pelvic pain during pregnancy: a descriptive study of signs and symptoms of 870 patients in primary care. AB - STUDY DESIGN: The authors conducted a cross-sectional analysis. OBJECTIVE: The objective of this study was to describe the signs and symptoms of pregnant women with pain and dysfunction in the pelvic area. SUMMARY OF BACKGROUND DATA: Pelvic pain during pregnancy is common with incidence rates of 48% to 56%. The exact cause of pelvic pain during pregnancy is still unclear. Also unclear is the association between physical examination and the occurrence of pelvic pain during pregnancy. METHODS: Pregnant women with pelvic pain were sent for treatment to 2 physical therapy practices between January 1997 and January 2002. A standardized clinical examination protocol and an extensive questionnaire were used to obtain relevant clinical and demographic characteristics. RESULTS: In total, 870 women were included. The average score of the overall severity of the complaints was 7.8 (scale 0-15). Main complaints were located around the sacroiliac joints (76.6%) and the pubic symphysis (57.2%). The area of pain was not related to positive signs in passive hip movements and sacroiliac tests. The highest positive test results were found for Patrick sign (71.7%), active straight leg raise (ASLR) test (66.4%), resisted adduction (54.4%), and passive hip abduction (36.9%). Overall severity of complaints was related to age, number of weeks pregnant, passive hip flexion and internal rotation, and swimming. The total explained variance of these factors was 15.9%. No relationship was found between overall complaints and sacroiliac tests, pelvic pain during a previous pregnancy, extremely painful or type of previous deliveries. CONCLUSIONS: Pregnant patients with pelvic pain show a considerable level of complaints. The overall severity of complaints is not related to previous peripartum pelvic pain or type of deliveries or to commonly used tests. Further study on the role of clinical examination, including passive flexion and internal rotation of the hip joints, is recommended. PMID- 15543076 TI - Occupational gonadal and embryo/fetal doses from fluoroscopically assisted surgical treatments of spinal disorders. AB - STUDY DESIGN: Simulation of lumbar spine fluoroscopy used during surgical treatments of spinal disorders on a humanoid phantom and monitoring of the scattered radiation levels. OBJECTIVES: To assess the potential of adverse effects to progeny due to the preconceptual and embryo/fetal exposure to ionizing radiation resulting from the parental occupational exposure to scattered radiation from lumbar fluoroscopy. SUMMARY OF BACKGROUND DATA: There are no available data on embryo/fetal doses resulting from maternal occupational exposure in the orthopedic theater. Besides, studies on staff gonadal doses from fluoroscopically assisted spine surgery are scarce and their results are not generally applicable. METHODS: Lumbar spine anterior-posterior and lateral fluoroscopy were performed on an anthropomorphic phantom. Scattered radiation within the orthopedic theater was recorded at the staff genitals and waist level. Gonadal, abdominal surface, and embryo/fetal doses normalized to the dose-area product specific to each projection were calculated. RESULTS: If the annual dose limits of occupational exposure are continuously exhausted for 10 years, the resulting radiogenic risk of congenital malformation in infants born to the orthopedic theater staff will be at least two orders of magnitude lower than the corresponding spontaneous probability. The occupational exposure of the pregnant mother bears a negligible contribution to the risk of hereditary effect on the newborn's progeny compared with the natural incidence rate. CONCLUSIONS: Radiogenic genetic and embryo/fetal risks resulting from occupational exposure due to fluoroscopically assisted surgical treatments of spinal disorders are well within tolerance levels provided that rigorous confinement to all pertinent occupational dose constraints is established. PMID- 15543077 TI - New technique in congenital scoliosis involving fixation to the pelvis after hemivertebra excision. AB - STUDY DESIGN: The authors conducted a retrospective study involving description of a new surgical technique. OBJECTIVES: The objective of this study was to describe and evaluate the efficacy of a new technique to stabilize the spine of young and small patients with congenital scoliosis after lumbosacral hemivertebra excision. SUMMARY OF BACKGROUND DATA: Congenital scoliosis develops as the result of anomalous vertebral development and asymmetric growth. The vertebral anomalies include congenital fusions and congenital hemivertebrae. With lumbosacral hemivertebrae at the lumbosacral junction, surgical excision followed by instrumentation to close the resulting space is frequently the best way to correct the deformity and obtain spinal balance. Surgical stabilization is required for preventing recurrent deformity and restoration of spinal balance. METHODS: A new surgical technique for spinal correction and stabilization in congenital scoliosis is described. The technique involves hemivertebra excision followed by fixation of the adjacent normal vertebra to the ilium with screws and cables. This technique allows efficient correction in both the coronal and sagittal planes. RESULTS: This construct was prospectively studied in 3 cases over a 3- to 5(1/2)-year period. All cases had solid fusion and well-balanced spine at latest follow up. CONCLUSIONS: This prospective study demonstrates a successful surgical technique for spinal stabilization after congenital hemivertebra excision in infants and young children with relatively small and soft bones when standard constructs like pedicle screws may not be optimal devices. Coronal and sagittal balance (restoration with improvement of lordosis) was successfully achieved in all cases. PMID- 15543078 TI - The profit motive and spine surgery. AB - STUDY DESIGN: The profit motive and market medicine have had a significant impact on clinical practice and research in the field of spine surgery. An overview of current concerns is presented. OBJECTIVE: The objective of this study was to provide those involved in the study and treatment of spinal disorders with a critical overview of the effects of the profit motive on our practices. SUMMARY OF BACKGROUND DATA: Historically, the profit motive has been viewed as eroding the standards of spine surgery, encouraging surgeons to operate aggressively and researchers to bias their results. Although there are legitimate concerns regarding the role played by such market forces, the profit motive exerts several quite positive effects on spine surgery as well. METHODS: Negative and positive aspects of the profit motive in spine surgery are explored along with alternative approaches. RESULTS: The profit motive in spine surgery can result in unnecessary surgery, as well as the push to market of unproven technologies. Yet, without a robust profit motive, it is unclear where sufficient funding could be found to support research and education, and to underwrite the advancement of new technologies. CONCLUSIONS: The profit motive significantly influences the way we practice and conduct research in spine surgery. To minimize the negative aspects of the profit motive, spine surgeons and researchers must refrain from being used by companies to rush products to market and/or compromising patient care out of self-interest. PMID- 15543079 TI - The coxib conundrum: lessons from the rise and fall of rofecoxib. PMID- 15543080 TI - The politics of therapeutics. PMID- 15543081 TI - A randomized, double-blind comparison of lercanidipine 10 and 20 mg in patients with stable effort angina: clinical evaluation of cardiac function by ambulatory ventricular scintigraphic monitoring. AB - We evaluated the antiischemic action and the effects on left ventricular response to exercise of lercanidipine, a long-acting dihydropyridine calcium antagonist, in 23 patients with stable effort angina in a randomized, double-blind, parallel trial. Left ventricular function was assessed during upright bicycle exercise using an ambulatory radionuclide detector for continuous noninvasive monitoring of cardiac function. Exercise was performed under control conditions before (run in placebo period) and after 2-week treatment with lercanidipine 10 or 20 mg once daily. During the placebo run-in period and at the study end, patients underwent clinical examination, ECG, exercise tests, ambulatory ventricular scintigraphic monitoring (VEST). Results showed that both drug doses increased time to onset of ST segment depression >/=1 mm and peak ST segment depression, with improvement of total exercise duration. Heart rate, blood pressure, and the rate-pressure product did not significantly change with respect to pretreatment value. The left ventricular ejection fraction, indicating contractility state of myocardium, was unchanged at rest and during exercise after both lercanidipine doses. In conclusion, lercanidipine is safe and effective in reducing ischemia in patients with stable effort angina without any deterioration of cardiac function. PMID- 15543082 TI - Cotransport of macrolide and fluoroquinolones, a beneficial interaction reversing P-glycoprotein efflux. AB - The purpose of this study was to determine the interactions of erythromycin and various fluoroquinolones with P-glycoprotein (P-gp) and in turn assess their effects on transport kinetics across a model cell monolayer. MDCKII-MDRI cells were selected as a model monolayer to evaluate the effects of various fluoroquinolones, ie, norfloxacin, lomefloxacin, ofloxacin, enoxacin, grepafloxacin, levofloxacin, and sparfloxacin on the P-gp-mediated efflux of H cyclosporine (CsA) and C-erythromycin. IC50 values associated with grepafloxacin , levofloxacin-, and sparfloxacin-mediated inhibition of P-gp were calculated across Caco-2 cells with erythromycin as the model P-gp substrate. Transport of erythromycin was then studied with P-gp saturable concentrations of fluoroquinolones. Western blot analysis was performed on Caco-2 cells to confirm P-gp expression. Only grepafloxacin elevated the uptake of H-CsA across the MDCKII-MDRI cell monolayer, whereas norfloxacin, lomefloxacin, ofloxacin, and enoxacin did not exert any effect on H-CsA uptake. Inhibition studies indicate that grepafloxacin, levofloxacin, and sparfloxacin are potent inhibitors of P-gp mediated efflux of C-erythromycin in the MDCKII-MDRI cell monolayer. Similar studies were conducted across Caco-2 cells and IC50 values were calculated. Inhibitory potency of sparfloxacin (IC50 = 607.6 microM) exceeded that of levofloxacin (IC50 = 1644 microM) and grepafloxacin (IC50 = 2266 microM). Permeability ratio (BL-AP/AP-BL) of C-erythromycin was found to be 8.67, which was reduced to 1.18, 1.83, and 1.39 in the presence of grepafloxacin (1 mmol/L), levofloxacin (5 mmol/L), and sparfloxacin (1 mmol/L), respectively. Log partition coefficient of grepafloxacin (1.58), levofloxacin (0.553), and sparfloxacin (0.45) were correlated with the inhibition of P-gp. Western blot analysis indicated the expression of P-gp in Caco-2 cells. Fluoroquinolones like grepafloxacin, levofloxacin, and especially sparfloxacin significantly inhibit the efflux of erythromycin, which can modulate oral absorption and disposition of macrolide drugs when administered concomitantly. PMID- 15543083 TI - Acetaminophen availability increases in Canada with no increase in the incidence of reports of inpatient hospitalizations with acetaminophen overdose and acute liver toxicity. AB - In September 1999, several Canadian provinces had place-of-sale restrictions lifted that had limited the sale of acetaminophen >325 mg and packages >24 tablets (any strength) to pharmacies only. This allowed the sale of all strengths of immediate-release acetaminophen in all package sizes in nonpharmacy locations. This study's purpose was to explore the effect that lifting restrictions on acetaminophen place of sale may have had on reported hospitalizations in Canada related to acetaminophen overdose toxicity. Using hospital discharge data, provinces with no preexisting restrictions on place of sale were compared with those in which restrictions were lifted in September 1999. Cases of reported APAP overdose included ICD-9/9-CM code 965.4, ICD-9 code E850.2, or ICD-9-CM code E850.4. Cases with reported acute liver toxicity included ICD-9/9-CM codes 570, 572.2, 572.4, V42.7, or procedure code 50.5. There were no significant differences between the 1.5-year periods pre- and post-September 1999 in annual incidence rates per 100,000 persons ages >/=12 years of hospitalizations reported with acetaminophen overdose, either overall or limited to those with death as an outcome, or in hospitalization reports with both acetaminophen overdose and acute liver toxicity, either overall (provinces with no restrictions: pre = 0.70, post = 0.80, P = 0.6328; provinces with restrictions lifted in September 1999: pre = 0.49, post = 0.47, P = 0.8649) or limited to those with death as an outcome (provinces with no restrictions: pre = 0.22, post = 0.12, P = 0.3030; provinces with restrictions lifted in September 1999: pre = 0.13, post = 0.09, P = 0.3589). In conclusion, the decision to lift Canadian place-of-sale restrictions increased acetaminophen availability and did not increase the rate of reported hospitalizations related to acetaminophen overdose toxicity. PMID- 15543084 TI - Examination of the cutaneous absorption of copper after the use of copper containing ointments. AB - Because copper-containing ointments are frequently used in anthroposophical medicine, a phase I trial to investigate the cutaneous absorption of copper was conducted. Sixty-one volunteers were randomized [group A: 0.4% copper (I) oxide, 13 men and 18 women (19-55 years); group B: 20% elementary copper, 11 men and 19 women (18-70 years)]. The ointment was applied over a 4-week period followed by a 4-week wash-out phase. Serum and urine copper concentrations were measured by atomic absorption spectrometry and hair copper concentration by inductive coupled plasma mass spectrometry. For statistical analysis, the Student t test for related random samples was used; alpha = 0.05 was chosen for the standard error. In group A, an increase of copper in serum and scalp hair and a decrease in urine were found in the study period. The mean serum concentration in all premenopausal women using oral contraceptives was above normal. In group B, the serum copper concentration increased significantly; in urine, it decreased, and in scalp hair, it remained stable. A higher level of serum copper was found in female volunteers using hormonal contraception. Treatment with the 2 different ointments did not cause toxic irritations on the skin, and it can therefore be deduced that the appropriate application of ointment preparations containing copper in concentrations up to 20% do not present a toxic risk. PMID- 15543085 TI - Inhibition of human plasma leucine5-enkephalin aminopeptidase hydrolysis by various endogenous peptides and a select number of clinically used drugs. AB - We identified a number of clinically used drugs and biologically active endogenous peptides able to significantly decrease the rate of human plasmatic aminopeptidase (AP) leucine-enkephalin (LEU) degradation. Bacitracin, bestatin, fluvoxamine, and each of 4 peptides tested significantly increased, in a dose dependent manner (10-10 M), LEU degradation half-life (t1/2) in each of 5 plasma samples studied. Each sample was obtained by pooling equal volume of 6 randomly selected, individual plasmas (4 male and 2 female healthy, drug-free volunteers). Thirty subjects (20 females and 10 males) participated in this study. With the exception of fluvoxamine, this inhibitory effect was lacking in various other commonly used drugs with widely different chemical structures and pharmacological profiles, eg, antidepressants (SSRIs, imipramine-like tricyclics, MAOIs), acute antimigraine agents (triptan class drugs), the nonselective beta-adrenergic antagonist propranolol, and serotonin receptor agonists and antagonists. Agents (concentration 10 M used as illustration), listed in decreasing order of LEU-AP inhibitory activity: substance P > angiotensin III > methionine-enkephalin > angiotensin II > fluvoxamine > bestatin gave t1/2 values (+/- SD) of 39.3 +/- 1.1, 29.4 +/- 0.8, 28.3 +/- 0.8, 27.4 +/- 0.7, 24.5 +/- 1.5, and 23.6 +/- 0.9 minutes, respectively. Control, bacitracin, and fluphenazine (known LEU-AP inhibitors were used for comparison) values of 11.8 +/- 1.0, 31.3 +/- 0.7, and 19.6 +/- 1.0 minutes, respectively. As expected, these drugs significantly decreased the initial velocity of peptide degradation; Iv values (+/- SD) of: 0.17 +/- 0.1 (0.02 +/- 0.01), 0.23 +/- 0.2 (0.02 +/- 0.01), 0.25 +/- 0.2 (0.02 +/ 0.01), 0.26 +/- 0.2 (0.03 +/- 0.01), 0.31 +/- 0.1 (0.03 +/- 0.01), and 0.33 +/- 0.1 (0.03 +/- 0.01), respectively; control, bacitracin, and fluphenazine: 1.10 +/ 0.3 (0.12 +/- 0.03), 0.20 +/- 0.1 (0.02 +/- 0.01), and 0.82 +/- 0.2 (0.08 +/- 0.02) pg LEU/min (pg LEU/mg protein/min), respectively. Results emphasize the selective nature of chemical structures required to significantly inhibit AP activity and provided information that could help the rational design of agents with high specificity in a biologic milieu containing multiple peptidases. In this case, targeted modulation of the bioavailability of LEU and other endogenous AP-degraded hormonal and nonhormonal peptides could be useful in the treatment of the pathophysiology associated with various disease conditions. Whether their development could find useful pharmacological applications remains to be explored. PMID- 15543087 TI - Guidelines for the diagnosis and management of heart failure. AB - Heart failure has reached epidemic proportions. Five million Americans have been diagnosed with heart failure, and this number is expected to double within the next 30 years. One million patients are hospitalized annually with decompensated heart failure, and half will be readmitted for recurrent symptoms within 6 months. Heart failure primarily affects the elderly and is the most common reason for hospitalization in this age group. The prognosis for heart failure is poor, worse than most malignancies. Heart failure accounts for 280,000 deaths annually, and the 5-year survival rate is less than 50% despite multiple therapeutic advances. Poor survival may be attributed in part to poor application of evidence based heart failure therapies and patient noncompliance. This review describes current guidelines for diagnosing and managing patients with heart failure. PMID- 15543088 TI - Management of decompensated heart failure. AB - Hospitalizations for heart failure have increased threefold during the past 3 decades, and this trend is expected to continue for the next 25 years. Heart failure now is the largest single expense for Medicare, and hospitalizations account for more than half of these costs. Most hospitals sustain financial losses with heart failure management because expenses exceed reimbursement. The hospital emergency department often is the initial encounter site for patients with new-onset heart failure, but most heart failure emergency department visits are for recurrent decompensation. The majority of these patients will be admitted to the hospital. Although accurate diagnosis and effective treatment are important for improving outcomes and lowering costs, there are no published guidelines for managing acutely decompensated heart failure. This review describes new diagnostic and management strategies, utilizing the emergency department observational unit as a triage area, that may decrease hospital length of stay, reduce hospital costs, and prevent readmissions. PMID- 15543089 TI - The pharmacologic treatment of heart failure. AB - Congestive heart failure is common, affecting 4-5 million American's with more than 500,000 new cases each year. A number of therapies have been proposed for the treatment of CHF; some have been found useful and some not. Digoxin therapy long a mainstay of therapy has been found beneficial, but lower doses and caution with its use in women has been recently advised based on new information. Converting enzyme inhibitors and beta blockers both add benefit when used in combination. The addition of aldosterone antagonists to the digoxin, diuretic, ACE, and beta-blocker combination appears to offer benefit and the use of the new less toxic eplerone will become more frequently employed in selected patients. Sudden death is also an important contributor to mortality in CHF patients. Use of a amiodarone and ICD's have both been reported to offer benefit. Selecting the optimum cost effective therapy is a challenge to those treating heart failure patients. ICD's are recommended in class II, III CHF with EF < 30 and amiodarone may be the therapy of choice in selected class IV patients and in patients with EF > 30%. PMID- 15543090 TI - Atrial fibrillation: focus on new therapeutic strategies: pharmacologic and nonpharmacologic approaches. PMID- 15543091 TI - Adverse reactions to selective cyclooxygenase-2 inhibitors (coxibs). AB - Coxibs, selective inhibitors of arachidonic acid cyclooxygenase-2, constitute a new, clinically useful therapeutic class of drugs that possess analgesic, antiinflammatory, and antipyretic properties. The drug inserts of prescription information presently provide a warning contraindicating their use in patients with aspirin-intolerant asthma and aspirin and NSAID hypersensitivity. However, according to investigations reported until now, most of those NSAID-sensitive individuals will tolerate coxibs. Careful oral challenge is useful for patient management since some cutaneous and systemic reactions of hypersensitivity, especially when sulfonamide-containing coxibs are employed, may occur. PMID- 15543092 TI - Evidence-based medical perspectives: the evolving role of PSA for early detection, monitoring of treatment response, and as a surrogate end point of efficacy for interventions in men with different clinical risk states for the prevention and progression of prostate cancer. AB - Following FDA approval and introduction into the clinic in the mid-1980s, PSA testing has become arguably the most versatile serum tumor marker in urologic oncology with clinical use for early detection (screening) of prostate cancer (PC), risk stratification for clinical staging, prognosis, intermediate biomarker for monitoring tumor recurrence, and more recently as an intermediate biomarker for assessing therapeutic response to antiandrogens, radiation therapy, and chemotherapy. PSA now routinely guides health care providers for the clinical management of PC over a wide range of clinical risk states for men at risk of PC, after local definitive therapy and after systemic therapy to prevent progression to metastatic bone disease, and to palliate men with hormone refractory prostate cancer (HRPC). To further assess the evidence that supports these clinical applications, this commentary reviews and critically evaluates the emerging body of new data focusing on several recently published seminal articles by D'Amico et al and Thompson et al, the new National Comprehensive Cancer Network 2004 recommendations for starting PSA testing at the age of 40 years old, the latest results from 2 phase 3 randomized, controlled trials of taxane-based regimens showing improved survival for men with HRPC, and the recent US FDA Public Workshop on Clinical Trial Endpoints in Prostate Cancer that helped to distill and synthesize the current state of the art and the progress toward validation of PSA metrics (eg, PSA velocity) as a surrogate end point (SE) for treatment efficacy with taxane-based regimens. Furthermore, several randomized, controlled chemoprevention trials in progress evaluating agents such as selenium and vitamin E in high-risk cohorts are well poised to confirm the validity of PSA as an SE for clinical efficacy for the prevention and progression of PC. Although there continues to be a need to validate better biomarkers before diagnosis of PC (more sensitive and specific) and after diagnosis to discern between indolent and aggressive forms of PC, it is very likely that some metric of PSA as a biomarker alone or as part of a panel of other serum proteomic markers or tissue-derived multiplex gene expression arrays will be around for years to come as a useful tool for risk stratification, early detection, prognosis, prediction, and as an SE of efficacy for prevention and treatment of PC. PMID- 15543093 TI - Polar hysteria: an expression of hypervitaminosis A. AB - Isotretinoin (Accutane) is a drug closely related to the chemical structure of Vitamin A. The pharmacology and toxicology of these two retinoids is similar enough to warrant comparison. Accutane is a powerful drug which its manufacturer, Roche, indicates is limited for severe recalcitrant nodular acne. This potency is also reflected in Accutane's well-known ability to produce severe birth defects if taken during pregnancy. Less well-known is the risk of this lipid soluble chemical to affect the Central Nervous System. Reports of intracranial hypertension, depression, and suicidal indeation with Accutane use have prompted an examination of this serious and life threatening potential. Though Roche has added a warning to its product label for signs of depression and suicidal ideation, this product is being overprescribed for all forms of acne, including mild cases and moderate acne that have not been treated with alternative medications, which have a lesser risk of depression and suicide. There is no contesting that this drug is effective at clearing up the most severe forms of acne, but the public must be informed of its proper, limited indication for use; depression and suicide can follow in patients with no prior history of psychiatric symptoms or suicide attempts. PMID- 15543094 TI - Possible pathomechanisms of sudden infant death syndrome: key role of chronic hypoxia, infection/inflammation states, cytokine irregularities, and metabolic trauma in genetically predisposed infants. AB - Chronic hypoxia, viral infections/bacterial toxins, inflammation states, biochemical disorders, and genetic abnormalities are the most likely trigger of sudden infant death syndrome (SIDS). Autopsy studies have shown increased pulmonary density of macrophages and markedly more eosinophils in the lungs accompanied by increased T and B lymphocytes. The elevated levels of immunoglobulins, about 20% more muscle in the pulmonary arteries, increased airway smooth muscle cells, and increased fetal hemoglobin and erythropoietin are evidence of chronic hypoxia before death. Other abnormal findings included mucosal immune stimulation of the tracheal wall, duodenal mucosa, and palatine tonsils, and circulating interferon. Low normal or higher blood levels of cortisol often with petechiae on intrathoracic organs, depleted maternal IgG antibodies to endotoxin core (EndoCAb) and early IgM EndoCAb triggered, partial deletions of the C4 gene, and frequent IL-10-592*A polymorphism in SIDS victims as well as possible hypoxia-induced decreased production of antiinflammatory, antiimmune, and antifibrotic cytokine IL-10, may be responsible for the excessive reactions to otherwise harmless infections. In SIDS infants, during chronic hypoxia and times of infection/inflammation, several proinflammatory cytokines are released in large quantities, sometimes also representing a potential source of tissue damage if their production is not sufficiently well controlled, eg, by pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP). These proinflammatory cytokines down-regulate gene expression of major cytochrome P-450 and/or other enzymes with the specific effects on mRNA levels, protein expression, and enzyme activity, thus affecting metabolism of several endogenous lipophilic substances, such as steroids, lipid soluble vitamins, prostaglandins, leukotrienes, thromboxanes, and exogenous substances. In SIDS victims, chronic hypoxia, TNF-alpha and other inflammatory cytokines, and arachidonic acid (AA) as well as n-3 polyunsaturated fatty acids (FA), stimulated and/or augmented superoxide generation by polymorphonuclear leukocytes, which contributed to tissue damage. Chronic hypoxia, increased amounts of nonheme iron in the liver and adrenals of these infants, enhanced activity of CYP2C9 regarded as the functional source of reactive oxygen species (ROS) in some endothelial cells, and nicotine accumulation in tissues also intensified production of ROS. These increased quantities of proinflammatory cytokines, ROS, AA, and nitric oxide (NO) also resulted in suppression of many CYP450 and other enzymes, eg, phosphoenolpyruvate carboxykinase (PEPCK), an enzyme important in the metabolism of FA during gluconeogenesis and glyceroneogenesis. PEPCK deficit found in SIDS infants (caused also by vitamin A deficiency) and eventually enhanced by PACAP lipolysis of adipocyte triglycerides resulted in an increased FA level in blood because of their impaired reesterification to triacylglycerol in adipocytes. In turn, the overproduction and release of FA into the blood of SIDS victims could lead to the metabolic syndrome and an early phase of type 2 diabetes. This is probably the reason for the secondary overexpression of the hepatic CYP2C8/9 content and activity reported in SIDS infants, which intensified AA metabolism. Pulmonary edema and petechial hemorrhages often present in SIDS victims may be the result of the vascular leak syndrome caused by IL-2 and IFN-alpha. Chronic hypoxia with the release of proinflammatory mediators IL-1alpha, IL-1beta and IL-6, and overloading of the cardiovascular and respiratory systems due to the narrowing airways and small pulmonary arteries of these children could also contribute to the development of these abnormalities. Moreover, chronic hypoxia of SIDS infants induced also production of hypoxia-inducible factor 1alpha (HIF-1alpha), which stimulated synthesis and release of different growth factors by vascular endothelial cells and intensified subclinical inflammatory reactions in the central nervous system, perhaps potentiated also by PACAP and VIP gene mutations. These processes could lead to the development of brainstem gliosis and disorders in the release of neuromediators important for physiologic sleep regulation. All these changes as well as eventual PACAP abnormalities could result in disturbed homeostatic control of the cardiovascular and respiratory responses of SIDS victims, which, combined with the nicotine effects and metabolic trauma, finally lead to death in these often genetically predisposed children. PMID- 15543095 TI - A wider perspective. PMID- 15543096 TI - Curry leaves. PMID- 15543097 TI - Rotation of wisdom teeth. PMID- 15543098 TI - Rofecoxib and dental prescribing. PMID- 15543099 TI - Dental understanding. PMID- 15543100 TI - Sugar-sweetened drinks. PMID- 15543108 TI - Endodontics: Part 6 Rubber dam and access cavities. AB - Rubber dam is easy to apply once the basic components and principles are understood. An efficient and well-trained dental nurse will greatly facilitate the application procedure. Although preparation of the access cavity may be commenced before rubber dam is applied to enable anatomical landmarks to be followed, the rubber dam should be placed as soon as possible with adequate protection against contamination of the access. The access cavity reflects the shape of the pulp chamber, modified by the angle of instrument approach. PMID- 15543109 TI - Facilitators and barriers to improving the quality of referrals for potential oral cancer. AB - The quality and content of referral letters are important for prioritisation of patients who may have oral cancer. Referrals letters to the Oral Medicine Clinic at Birmingham Dental Hospital were analysed and practitioners interviewed. Whilst acceptable for general purposes, most letters did not contain sufficient information to allow effective prioritisation. Interviews disclosed a misunderstanding amongst practitioners about the way in which referrals were handled. A number of barriers to increasing the information included in letters were identified. Referral guidelines and a standardised proforma might help improve the ability of the service to operate a fast-track system. PMID- 15543110 TI - An easy way of intruding an upper central incisor. AB - Discrepancies of the incisal edges of the anterior teeth, particularly in the upper jaw, could adversely affect dental aesthetics. This paper presents an easy and straight forward treatment of such a case, which was accomplished taking the patient's demands and profession into consideration. In dentistry, the social status of the patient could affect the treatment plan. PMID- 15543117 TI - An RCT pilot study to test the effects of intravenous midazolam as a conscious sedation technique for anxious children requiring dental treatment--an alternative to general anaesthesia. AB - AIM: To add to the evidence base for acceptable and effective paediatric conscious sedation techniques in dental primary care. OBJECTIVES: To compare three conscious sedation techniques for primary care as an alternative to dental general anaesthesia (DGA) in children. To assess the feasibility and practicality of running the trial in general dental practice. To form the basis for sample size calculations and assess scales of measurement. DESIGN: Single centre, randomised control trial (RCT). SETTING: Queensway Anxiety Management Clinic (QAMC). A primary care based general and referral dental practice for the management of anxious patients. SUBJECTS, MATERIALS AND METHODS: Sixty five children too anxious for management with relative analgesia, requiring invasive dental procedure for which dental general anaesthesia (DGA) will be required if an alternative cannot be found. INTERVENTIONS: Group 1 (n = 20) - A combination of inhaled medical air and titrated intravenous midazolam. Group 2 (n = 22) - A combination of inhaled 40% nitrous oxide in oxygen and titrated intravenous midazolam. Group 3 (n = 23) - A combination of an inhaled mixture of 0.3% sevoflurane and 40% nitrous oxide in oxygen with titrated intravenous midazolam. MAIN OUTCOME MEASURES: Successful completion of the intended dental treatment with a child who is co-operative and responsive to verbal commands. RESULTS: Fifty per cent (ten children) successfully completed treatment in Group 1, 73% (16 children) in Group 2 and 83% (19 children) in Group 3. This difference was not significant at a 5% level (chi(2) = 5.53, df = 2, P = 0.07) Of the total of 20 failures, eight children in Group 1 and one child in Group 2 were successfully treated with the addition of sevoflurane and nitrous oxide in oxygen. Only two children required referral to a hospital setting for DGA and the remaining nine children were managed with an alternative conscious sedation technique. CONCLUSION: This pilot shows that intravenous midazolam especially in combination with the addition of inhaled nitrous oxide or sevoflurane and nitrous oxide were promising safe and effective techniques, sufficient to justify progression to a definitive RCT with appropriate methods. PMID- 15543118 TI - A study to assess the efficacy of a new detergent free, whitening dentifrice in vivo using QLF planimetric analysis. AB - OBJECTIVE: To determine the effects of a detergent-free, whitening dentifrice using an in vivo plaque regrowth model with the novel application of QLF as a planimetric analysis tool. METHOD: A total of 20 subjects took part in a double blind, single-centre, crossover study in which slurry rinses were the only form of plaque control over a 5-day period. Following a washout and prophylaxis the subjects used 2 daily rinses in the absence of all other plaque control methods. Subjects returned to the clinic on the afternoon of day 5 when plaque was disclosed and assessed by the plaque index and area using both a photographic and novel fluorescent planimetric technique. A further 9-day washout was carried out and the rinse period repeated to ensure that each subject had used both experimental and comparator slurries. RESULTS: Twenty subjects completed the trial. The test product showed a significant inhibition of plaque re-growth (16.9%) compared with a fluoride-matched comparator using the Turesky index (P < 0.0001), the photographic planimetric technique (17.5%) (P < 0.0001) and the novel QLF technique (18.4%) (P < 0.0001). CONCLUSION: The results confirm that plaque inhibition capability of a detergent-free whitening dentifrice is at least as effective as a fluoride matched comparator. QLF is a promising tool for disclosed plaque quantification. PMID- 15543119 TI - The future of the profession--a survey of dental school applicants. AB - PURPOSE: To gather background information about applicants to the dental undergraduate course, and ascertain the factors involved in their decision to study dentistry. METHOD: Applicants attending for interview at Dundee and Manchester Dental Schools completed anonymous questionnaires. The useable response rate was 94% (n = 436). RESULTS: There were equal numbers of male and female applicants to the two schools as a whole. Although there was a much broader ethnic mix in the applicants to Manchester, the overall proportion of minority ethnic groups was considerably greater than in the UK population as a whole. Seventy-five per cent came from professional backgrounds, although marked differences were apparent between ethnic groups. A quarter had family relations who were dentists, and 30% were related to doctors. Over half (53%) decided to apply within the last year, with popular motivating factors including a general interest in dentistry, a desire to help people and to work with their hands. Dentistry was the first choice of career of 89%, with medicine the most popular alternative. CONCLUSION: Valuable information was gathered from these prospective dental students regarding their backgrounds and the decision process involved in their career selection. This will prove beneficial when developing the recruitment process and allow appropriate tailoring to the target audience. PMID- 15543120 TI - In sickness and in health. AB - This final instalment in the literary series deals with the relationship between general and oral health and methods of maintaining good oral hygiene. PMID- 15543132 TI - Mast cells promote homeostasis by limiting endothelin-1-induced toxicity. AB - Endothelin-1 (ET-1) is a 21-amino-acid peptide, derived from vascular endothelial cells, with potent vasoconstrictor activity. ET-1 has been implicated in diverse physiological or pathological processes, including the vascular changes associated with sepsis. However, the factors that regulate ET-1-associated toxicity during bacterial infections, or in other settings, are not fully understood. Both the pathology associated with certain allergic and autoimmune disorders, and optimal host defence against bacterial and parasitic infections are mediated by mast cells. In vitro, mast cells can produce ET-1 (ref. 11), undergo ET-1-dependent and endothelin-A receptor (ET(A))-dependent activation, and release proteases that degrade ET-1 (ref. 14). Although the potential relationships between mast cells and the ET-1 system thus may be complex, the importance of interactions between ET-1 and mast cells in vivo is obscure. Here we show that ET(A)-dependent mast-cell activation can diminish both ET-1 levels and ET-1-induced pathology in vivo, and also can contribute to optimal survival during acute bacterial peritonitis. These findings identify a new biological function for mast cells: promotion of homeostasis by limiting the toxicity associated with an endogenous mediator. PMID- 15543133 TI - Complete sequence and comparative genome analysis of the dairy bacterium Streptococcus thermophilus. AB - The lactic acid bacterium Streptococcus thermophilus is widely used for the manufacture of yogurt and cheese. This dairy species of major economic importance is phylogenetically close to pathogenic streptococci, raising the possibility that it has a potential for virulence. Here we report the genome sequences of two yogurt strains of S. thermophilus. We found a striking level of gene decay (10% pseudogenes) in both microorganisms. Many genes involved in carbon utilization are nonfunctional, in line with the paucity of carbon sources in milk. Notably, most streptococcal virulence-related genes that are not involved in basic cellular processes are either inactivated or absent in the dairy streptococcus. Adaptation to the constant milk environment appears to have resulted in the stabilization of the genome structure. We conclude that S. thermophilus has evolved mainly through loss-of-function events that remarkably mirror the environment of the dairy niche resulting in a severely diminished pathogenic potential. PMID- 15543134 TI - RNAi-mediated replacement of morphine with the nonnarcotic alkaloid reticuline in opium poppy. AB - We report on the silencing of codeinone reductase (COR) in the opium poppy, Papaver somniferum, using a chimeric hairpin RNA construct designed to silence all members of the multigene COR family through RNA interference (RNAi). After gene silencing, the precursor alkaloid (S)-reticuline-seven enzymatic steps upstream of codeinone-accumulated in transgenic plants at the expense of morphine, codeine, oripavine and thebaine. Methylated derivatives of reticuline also accumulated. Analysis verified loss of Cor gene transcript, appearance of 22 mer degradation products and reduction of enzyme activity. The surprising accumulation of (S)-reticuline suggests a feedback mechanism preventing intermediates from general benzylisoquinoline synthesis entering the morphine specific branch. However transcript levels for seven other enzymes in the pathway, both before and after (S)-reticuline, were unaffected. This is the first report of gene silencing in transgenic opium poppy and of metabolic engineering to cause the high-yield accumulation of the nonnarcotic alkaloid reticuline. PMID- 15543135 TI - Rab27A-binding protein Slp2-a is required for peripheral melanosome distribution and elongated cell shape in melanocytes. AB - The synaptotagmin-like protein (Slp) family is implicated in regulating Rab27A mediated membrane transport, but how it might do this is unknown. Here we report that Slp2-a, a previously uncharacterized Rab27A-binding protein in melanocytes, controls melanosome distribution in the cell periphery and regulates the morphology of melanocytes. Slp2-a is the most abundantly expressed of the Slp- and Slac2-family proteins in melanocytes and colocalizes with Rab27A on melanosomes. Knockdown of endogenous Slp2-a protein by small-interfering RNAs (siRNAs) markedly reduced the number of melanosomes in the cell periphery of mouse melanocytes ('peripheral dilution'). Expression of siRNA-resistant Slp2-a (Slp2-a(SR)) rescued the peripheral dilution of melanosomes induced by Slp2-a siRNAs, but Slp2-a(SR) mutants, which failed to interact with either phospholipids or Rab27A, did not. Loss of Slp2-a protein also induced a change in melanocyte morphology, from their normal elongated shape to a more rounded shape, which depended on the phospholipid-binding activity of Slp2-a, but not on its Rab27A-binding activity. By contrast, knockdown of Slac2-a (also called melanophilin), another Rab27A-binding protein in melanocytes, caused perinuclear aggregation of melanosomes alone without altering cell shape. These results reveal the differential and sequential roles of Rab27A-binding proteins in melanosome transport in melanocytes. PMID- 15543136 TI - The Williams syndrome transcription factor interacts with PCNA to target chromatin remodelling by ISWI to replication foci. AB - Chromatin states have to be faithfully duplicated during DNA replication to maintain cell identity. It is unclear whether or how ATP-dependent chromatin remodelling factors are involved in this process. Here we provide evidence that the Williams syndrome transcription factor (WSTF) is targeted to replication foci through direct interaction with the DNA clamp PCNA, an important coordinator of DNA and chromatin replication. WSTF, in turn, recruits imitation switch (ISWI) type nucleosome-remodelling factor SNF2H to replication sites. These findings reveal a novel recruitment mechanism for ATP-dependent chromatin-remodelling factors that is fundamentally different from the previously documented targeting by sequence-specific transcriptional regulators. RNA-interference-mediated depletion of WSTF or SNF2H causes a compaction of newly replicated chromatin and increases the amount of heterochromatin markers, including HP1beta. This increase in the amount of HP1beta protein is mediated by progression through S phase and is not the result of an increase in HP1beta mRNA levels. We propose that the WSTF ISWI complex has a role in the maintenance of chromatin structures during DNA replication. PMID- 15543137 TI - Guidance of myocardial patterning in cardiac development by Sema6D reverse signalling. AB - Cardiac chamber formation involves dynamic changes in myocardial organization, including trabeculation and expansion of the compact layer. The positional cues that regulate myocardial patterning, however, remain unclear. Through ligation of the Plexin-A1 receptor, the transmembrane-type semaphorin Sema6D regulates endocardial cell migration. Here, we demonstrate that knockdown of either Sema6D or Plexin-A1 leads to the generation of a small, thin ventricular compact layer and to defective trabeculation. In the heart, expression of the Plexin-A1 extracellular domain alone can rescue the defective trabeculation induced by suppression of Plexin-A1, but not that resulting from defective Sema6D expression. This indicates that reverse signalling by Sema6D occurs within the myocardium. In a ligand-dependent manner, Abl kinase is recruited to the cytoplasmic tail of Sema6D and activated, resulting in phosphorylation of Enabled and dissociation from Sema6D. Constitutive activation of Sema6D signalling enhances the migration of myocardial cells into the trabeculae, whereas inhibition arrests cells within the compact layer. Thus, Sema6D coordinates both compact-layer expansion and trabeculation, functioning as both a ligand and a receptor for Plexin-A1. PMID- 15543138 TI - Non-canonical Wnt signals are modulated by the Kaiso transcriptional repressor and p120-catenin. AB - Gastrulation movements are critical for establishing the three principal germ layers and the basic architecture of vertebrate embryos. Although the individual molecules and pathways involved are not clearly understood, non-canonical Wnt signals are known to participate in developmental processes, including planar cell polarity and directed cell rearrangements. Here we demonstrate that the dual specificity transcriptional repressor Kaiso, first identified in association with p120-catenin, is required for Xenopus gastrulation movements. In addition, depletion of xKaiso results in increased expression of the non-canonical xWnt11, which contributes to the xKaiso knockdown phenotype as it is significantly rescued by dominant-negative Wnt11. We further demonstrate that xWnt11 is a direct gene target of xKaiso and that p120-catenin association relieves xKaiso repression in vivo. Our results indicate that p120-catenin and Kaiso are essential components of a new developmental gene regulatory pathway that controls vertebrate morphogenesis. PMID- 15543139 TI - Selective reconfiguration of layer 4 visual cortical circuitry by visual deprivation. AB - Visual deprivation during a developmental sensitive period markedly alters visual cortical response properties, but the changes in intracortical circuitry that underlie these effects are poorly understood. Here we use a slice preparation of rat primary visual cortex to show that 2 d of prior visual deprivation early in life increases the excitability of layer 4 circuitry. Slice recordings showed that spontaneous activity of layer 4 star pyramidal neurons increased 25-fold after 2 d of visual deprivation between postnatal days (P) 15 and P17. This effect was mediated by increased net excitatory and decreased net inhibitory synaptic drive. Paired recordings showed that excitatory connections between star pyramidal neurons doubled in amplitude, whereas inhibitory connections decreased or increased depending on the interneuron class. These effects reversed when vision was restored. This dynamic adjustment of the excitation-inhibition balance may allow the networks within layer 4 to maintain stable levels of activity in the face of variable sensory input. PMID- 15543140 TI - Zebrafish unplugged reveals a role for muscle-specific kinase homologs in axonal pathway choice. AB - En route to their target, pioneering motor growth cones repeatedly encounter choice points at which they make pathway decisions. In the zebrafish mutant unplugged, two of the three segmental motor axons make incorrect decisions at a somitic choice point. Using positional cloning, we show here that unplugged encodes a homolog of muscle-specific kinase (MuSK) and that, unlike mammalian MuSK, unplugged has only a limited role in neuromuscular synaptogenesis. We demonstrate that unplugged is transiently expressed in cells adjacent to the choice point and that unplugged signaling before the arrival of growth cones induces changes in the extracellular environment. In addition, we find that the unplugged locus generates three different transcripts. The splice variant 1 (SV1) isoform lacks the extracellular modules essential for agrin responsiveness, and signaling through this isoform mediates axonal pathfinding, independent of the MuSK downstream component rapsyn. Our results demonstrate a new role for MuSK homologs in axonal pathway selection. PMID- 15543141 TI - Math1 regulates development of the sensory epithelium in the mammalian cochlea. AB - The transcription factor Math1 (encoded by the gene Atoh1, also called Math1) is required for the formation of mechanosensory hair cells in the inner ear; however, its specific molecular role is unknown. Here we show that absence of Math1 in mice results in a complete disruption of formation of the sensory epithelium of the cochlea, including the development of both hair cells and associated supporting cells. In addition, ectopic expression of Math1 in nonsensory regions of the cochlea is sufficient to induce the formation of sensory clusters that contain both hair cells and supporting cells. The formation of these clusters is dependent on inhibitory interactions mediated, most probably, through the Notch pathway, and on inductive interactions that recruit cells to develop as supporting cells through a pathway independent of Math1. These results show that Math1 functions in the developing cochlea to initiate both inductive and inhibitory signals that regulate the overall formation of the sensory epithelia. PMID- 15543142 TI - Adapter protein 14-3-3 is required for a presynaptic form of LTP in the cerebellum. AB - Long-term potentiation (LTP) of granule cell-Purkinje cell synapses in the mouse cerebellum requires phosphorylation by protein kinase A of the active-zone protein RIM1alpha at Ser413. Here, we show that the adapter protein 14-3-3 readily binds phosphorylated Ser413 in RIM1alpha, and that presynaptic transfection with a dominant-negative 14-3-3eta mutant, or a RIM1alpha mutant with enhanced 14-3-3 binding, inhibits LTP. Thus, RIM1alpha phosphorylation triggers presynaptic LTP in part through recruitment of 14-3-3 to phospho-Ser413 RIM1alpha. PMID- 15543143 TI - The neurotoxic MEC-4(d) DEG/ENaC sodium channel conducts calcium: implications for necrosis initiation. AB - Hyperactivation of the Caenorhabditis elegans MEC-4 Na(+) channel of the DEG/ENaC superfamily (MEC-4(d)) induces neuronal necrosis through an increase in intracellular Ca(2+) and calpain activation. How exacerbated Na(+) channel activity elicits a toxic rise in cytoplasmic Ca(2+), however, has remained unclear. We tested the hypothesis that MEC-4(d)-induced membrane depolarization activates voltage-gated Ca(2+) channels (VGCCs) to initiate a toxic Ca(2+) influx, and ruled out a critical requirement for VGCCs. Instead, we found that MEC-4(d) itself conducts Ca(2+) both when heterologously expressed in Xenopus oocytes and in vivo in C. elegans touch neurons. Data generated using the Ca(2+) sensor cameleon suggest that an induced release of endoplasmic reticulum (ER) Ca(2+) is crucial for progression through necrosis. We propose a refined molecular model of necrosis initiation in which Ca(2+) influx through the MEC 4(d) channel activates Ca(2+)-induced Ca(2+) release from the ER to promote neuronal death, a mechanism that may apply to neurotoxicity associated with activation of the ASIC1a channel in mammalian ischemia. PMID- 15543144 TI - Baz, Par-6 and aPKC are not required for axon or dendrite specification in Drosophila. AB - Par-3/Baz, Par-6, and aPKC are evolutionarily conserved regulators of cell polarity, and overexpression experiments implicate them as axon determinants in vertebrate hippocampal neurons. Here we examined their mutant and overexpression phenotypes in Drosophila melanogaster. We found that mutants neurons had normal axon and dendrite morphology and remodeled axons correctly in metamorphosis, and that overexpression did not affect axon or dendrite specification. Baz/Par-6/aPKC are therefore not essential for axon specification in Drosophila. PMID- 15543145 TI - Receptor tyrosine kinase ErbB4 modulates neuroblast migration and placement in the adult forebrain. AB - Neural progenitor proliferation, differentiation and migration are continually active in the rostral migratory stream of the adult brain. Here, we show that the receptor tyrosine kinase ErbB4 is expressed prominently by the neuroblasts present in the subventricular zone and the rostral migratory stream. The neuregulins (NRG1-NRG3), which have been identified as ErbB4 ligands, are detected either in the stream or in adjacent regions. Mice deficient in ErbB4 expressed under the control of either the nestin or the hGFAP promoter have altered neuroblast chain organization and migration and deficits in the placement and differentiation of olfactory interneurons. These findings suggest that ErbB4 activation helps to regulate the organization of neural chains that form the rostral migratory stream and influences the differentiation of olfactory interneuronal precursors. PMID- 15543146 TI - Mutations in PTF1A cause pancreatic and cerebellar agenesis. AB - Individuals with permanent neonatal diabetes mellitus usually present within the first three months of life and require insulin treatment. We recently identified a locus on chromosome 10p13-p12.1 involved in permanent neonatal diabetes mellitus associated with pancreatic and cerebellar agenesis in a genome-wide linkage search of a consanguineous Pakistani family. Here we report the further linkage analysis of this family and a second family of Northern European descent segregating an identical phenotype. Positional cloning identified the mutations 705insG and C886T in the gene PTF1A, encoding pancreas transcription factor 1alpha, as disease-causing sequence changes. Both mutations cause truncation of the expressed PTF1A protein C-terminal to the basic-helix-loop-helix domain. Reporter-gene studies using a minimal PTF1A deletion mutant indicate that the deleted region defines a new domain that is crucial for the function of this protein. PTF1A is known to have a role in mammalian pancreatic development, and the clinical phenotype of the affected individuals implicated the protein as a key regulator of cerebellar neurogenesis. The essential role of PTF1A in normal cerebellar development was confirmed by detailed neuropathological analysis of Ptf1a(-/-) mice. PMID- 15543147 TI - 'Racial' differences in genetic effects for complex diseases. AB - 'Racial' differences are frequently debated in clinical, epidemiological and molecular research and beyond. In particular, there is considerable controversy regarding the existence and importance of 'racial' differences in genetic effects for complex diseases influenced by a large number of genes. An important question is whether ancestry influences the impact of each gene variant on the disease risk. Here, we addressed this question by examining the genetic effects for 43 validated gene-disease associations across 697 study populations of various descents. The frequencies of the genetic marker of interest in the control populations often (58%) showed large heterogeneity (statistical variability) between 'races'. Conversely, we saw large heterogeneity in the genetic effects (odds ratios) between 'races' in only 14% of cases. Genetic markers for proposed gene-disease associations vary in frequency across populations, but their biological impact on the risk for common diseases may usually be consistent across traditional 'racial' boundaries. PMID- 15543148 TI - Rapid analysis of the DNA-binding specificities of transcription factors with DNA microarrays. AB - We developed a new DNA microarray-based technology, called protein binding microarrays (PBMs), that allows rapid, high-throughput characterization of the in vitro DNA binding-site sequence specificities of transcription factors in a single day. Using PBMs, we identified the DNA binding-site sequence specificities of the yeast transcription factors Abf1, Rap1 and Mig1. Comparison of these proteins' in vitro binding sites with their in vivo binding sites indicates that PBM-derived sequence specificities can accurately reflect in vivo DNA sequence specificities. In addition to previously identified targets, Abf1, Rap1 and Mig1 bound to 107, 90 and 75 putative new target intergenic regions, respectively, many of which were upstream of previously uncharacterized open reading frames. Comparative sequence analysis indicated that many of these newly identified sites are highly conserved across five sequenced sensu stricto yeast species and, therefore, are probably functional in vivo binding sites that may be used in a condition-specific manner. Similar PBM experiments should be useful in identifying new cis regulatory elements and transcriptional regulatory networks in various genomes. PMID- 15543149 TI - A transcriptomic analysis of the phylum Nematoda. AB - The phylum Nematoda occupies a huge range of ecological niches, from free-living microbivores to human parasites. We analyzed the genomic biology of the phylum using 265,494 expressed-sequence tag sequences, corresponding to 93,645 putative genes, from 30 species, including 28 parasites. From 35% to 70% of each species' genes had significant similarity to proteins from the model nematode Caenorhabditis elegans. More than half of the putative genes were unique to the phylum, and 23% were unique to the species from which they were derived. We have not yet come close to exhausting the genomic diversity of the phylum. We identified more than 2,600 different known protein domains, some of which had differential abundances between major taxonomic groups of nematodes. We also defined 4,228 nematode-specific protein families from nematode-restricted genes: this class of genes probably underpins species- and higher-level taxonomic disparity. Nematode-specific families are particularly interesting as drug and vaccine targets. PMID- 15543150 TI - Visualizing dendritic cell networks in vivo. AB - In the steady state, dendritic cells (DCs) in the lymph node induce T cell tolerance to self antigens. Innate signals trigger the maturation of tissue DCs, which migrate into lymph nodes and activate T cells. To examine DCs in vivo, we produced transgenic mice whose DCs expressed enhanced yellow fluorescent protein. Two-photon microscopy of lymph nodes in live mice showed that most of the steady state DCs were enmeshed in an extensive network and remained in place while actively probing adjacent T cells with their processes. Mature DCs were more motile than steady-state DCs and were rapidly dispersed and integrated into the sessile network, facilitating their interaction with migrating T cells. PMID- 15543151 TI - Anisotropic lattice distortions in biogenic aragonite. AB - Composite biogenic materials produced by organisms have a complicated design on a nanometre scale. An outstanding example of organic-inorganic composites is provided by mollusc seashells, whose superior mechanical properties are due to their multi-level crystalline hierarchy and the presence of a small amount (0.1-5 wt%) of organic molecules. The presence of organic molecules, among other characteristics, can influence the coherence length for X-ray scattering in biogenic crystals. Here we show the results of synchrotron high-resolution X-ray powder diffraction measurements in biogenic and non-biogenic (geological) aragonite crystals. On applying the Rietveld refinement procedure to the high resolution diffraction spectra, we were able to extract the aragonite lattice parameters with an accuracy of 10 p.p.m. As a result, we found anisotropic lattice distortions in biogenic aragonite relative to the geological sample, maximum distortion being 0.1% along the c axis of the orthorhombic unit cell. The organic molecules could be a source of these structural distortions in biogenic crystals. This finding may be important to the general understanding of the biomineralization process and the development of bio-inspired 'smart' materials. PMID- 15543152 TI - Atomic dynamics and Marangoni films during liquid-metal spreading. AB - Despite its apparent simplicity, spreading of liquid metals at high temperatures has defied description and generalization. Wetting at high temperature is usually accompanied by interdiffusion and chemical reaction, but the forces that drive reactive spreading and the mechanisms that control its kinetics have been very poorly understood. The unsolved challenge has been to link macroscopic measurements such as the dynamic contact angle or the speed of a moving liquid front to phenomena occurring at the microscopic and even atomic level in the vicinity of the triple solid-liquid-vapour junction. We have taken a big step towards meeting this challenge. Our systematic analysis of the spreading of metal metal systems with varying degrees of mutual solubility allows us to report on the fundamental differences between the mechanisms controlling spreading of organic liquids and liquid metals and on formation of Marangoni films driven by surface-tension gradients in high-temperature systems. PMID- 15543153 TI - Mice lacking calsarcin-1 are sensitized to calcineurin signaling and show accelerated cardiomyopathy in response to pathological biomechanical stress. AB - Signaling by the calcium-dependent phosphatase calcineurin profoundly influences the growth and gene expression of cardiac and skeletal muscle. Calcineurin binds to calsarcins, a family of muscle-specific proteins of the sarcomeric Z-disc, a focal point in the pathogenesis of human cardiomyopathies. We show that calsarcin 1 negatively modulates the functions of calcineurin, such that calcineurin signaling was enhanced in striated muscles of mice that do not express calsarcin 1. As a consequence of inappropriate calcineurin activation, mice with a null mutation in calsarcin-1 showed an excess of slow skeletal muscle fibers. The absence of calsarcin-1 also activated a hypertrophic gene program, despite the absence of hypertrophy, and enhanced the cardiac growth response to pressure overload. In contrast, cardiac adaptation to other hypertrophic stimuli, such as chronic catecholamine stimulation or exercise, was not affected. These findings show important roles for calsarcins as modulators of calcineurin signaling and the transmission of a specific subset of stress signals leading to cardiac remodeling in vivo. PMID- 15543154 TI - U2-U6 RNA folding reveals a group II intron-like domain and a four-helix junction. AB - Intron removal in nuclear precursor mRNA is catalyzed through two transesterification reactions by a multi-megaDalton ribonucleoprotein machine called the spliceosome. A complex between U2 and U6 small nuclear RNAs is a core component of the spliceosome. Here we present an NMR structural analysis of a protein-free U2-U6 complex from Saccharomyces cerevisiae. The observed folding of the U2-U6 complex is a four-helix junction, in which the catalytically important AGC triad base-pairs only within U6 and not with U2. The base-pairing of the AGC triad extends the U6 intramolecular stem-loop (U6 ISL), and the NMR structure of this extended U6 ISL reveals structural similarities with domain 5 of group II self-splicing introns. The observed conformation of the four-helix junction could be relevant to the first, but not the second, step of splicing and may help to position the U6 ISL adjacent to the 5' splice site. PMID- 15543155 TI - Human CD59 is a receptor for the cholesterol-dependent cytolysin intermedilysin. AB - Cholesterol is believed to serve as the common receptor for the cholesterol dependent cytolysins (CDCs). One member of this toxin family, Streptococcus intermedius intermedilysin (ILY), exhibits a narrow spectrum of cellular specificity that is seemingly inconsistent with this premise. We show here that ILY, via its domain 4 structure, binds to the glycosyl-phosphatidylinositol linked membrane protein human CD59 (huCD59). CD59 is an inhibitor of the membrane attack complex of human complement. ILY specifically binds to huCD59 via residues that are the binding site for the C8alpha and C9 complement proteins. These studies provide a new model for the mechanism of cellular recognition by a CDC. PMID- 15543156 TI - Hsp70 and Hsp40 attenuate formation of spherical and annular polyglutamine oligomers by partitioning monomer. AB - Protein conformational changes that result in misfolding, aggregation and amyloid fibril formation are a common feature of many neurodegenerative disorders. Studies with beta-amyloid (Abeta), alpha-synuclein and other amyloid-forming proteins indicate that the assembly of misfolded protein conformers into fibrils is a complex process that may involve the population of metastable spherical and/or annular oligomeric assemblies. Here, we show by atomic force microscopy that a mutant huntingtin fragment with an expanded polyglutamine repeat forms spherical and annular oligomeric structures reminiscent of those formed by Abeta and alpha-synuclein. Notably, the molecular chaperones Hsp70 and Hsp40, which are protective in animal models of neurodegeneration, modulate polyglutamine aggregation reactions by partitioning monomeric conformations and disfavoring the accretion of spherical and annular oligomers. PMID- 15543157 TI - Structures of human MAP kinase kinase 1 (MEK1) and MEK2 describe novel noncompetitive kinase inhibition. AB - MEK1 and MEK2 are closely related, dual-specificity tyrosine/threonine protein kinases found in the Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) signaling pathway. Approximately 30% of all human cancers have a constitutively activated MAPK pathway, and constitutive activation of MEK1 results in cellular transformation. Here we present the X-ray structures of human MEK1 and MEK2, each determined as a ternary complex with MgATP and an inhibitor to a resolution of 2.4 A and 3.2 A, respectively. The structures reveal that MEK1 and MEK2 each have a unique inhibitor-binding pocket adjacent to the MgATP-binding site. The presence of the potent inhibitor induces several conformational changes in the unphosphorylated MEK1 and MEK2 enzymes that lock them into a closed but catalytically inactive species. Thus, the structures reported here reveal a novel, noncompetitive mechanism for protein kinase inhibition. PMID- 15543158 TI - Redox regulation of OxyR requires specific disulfide bond formation involving a rapid kinetic reaction path. AB - The Escherichia coli OxyR transcription factor is activated by cellular hydrogen peroxide through the oxidation of reactive cysteines. Although there is substantial evidence for specific disulfide bond formation in the oxidative activation of OxyR, the presence of the disulfide bond has remained controversial. By mass spectrometry analyses and in vivo labeling assays we found that oxidation of OxyR in the formation of a specific disulfide bond between Cys199 and Cys208 in the wild-type protein. In addition, using time-resolved kinetic analyses, we determined that OxyR activation occurs at a rate of 9.7 s( 1). The disulfide bond-mediated conformation switch results in a metastable form that is locally strained by approximately 3 kcal mol(-1). On the basis of these observations we conclude that OxyR activation requires specific disulfide bond formation and that the rapid kinetic reaction path and conformation strain, respectively, drive the oxidation and reduction of OxyR. PMID- 15543159 TI - Estimated change in physical activity level (PAL) and prediction of 5-year weight change in men: the Aerobics Center Longitudinal Study. AB - OBJECTIVE: To determine the relation between the average daily physical activity level (PAL) and the trajectory of weight change in men at risk for weight gain. DESIGN AND SETTING: Clinic-based cohort study over an average of 5 y. SUBJECTS: Healthy men (N=2501) ages 20-55 y participating in the Aerobics Center Longitudinal Study who had received at least four medical examinations at the Cooper Clinic between 1970 and 1998. MEASUREMENTS: Daily leisure-time physical activity was reported and body weight was measured at all four examinations. The average daily PAL (METs 24 h(-1)) was estimated from all activities, as well as from other incidental active and passive activities. Weight change over four examinations was regressed on the change in PAL between the first and third examinations. RESULTS: Random coefficient regression modeling indicated a curvilinear slope for weight gain over the follow-up among those maintaining the same PAL between the first and third examinations. Weight gain was further accelerated among men who decreased their activity. A shift from a low PAL (<1.45 METs 24 h(-1)) to a moderate (1.45-1.60 METs 24 h(-1)) or high (>1.60 METs 24 h( 1)) PAL was necessary for weight loss over time. Men with initially the lowest PAL had the greatest benefit from increasing activity. CONCLUSIONS: Daily PAL was inversely related to weight gain in this cohort. Increasing to or maintaining a daily PAL at least 60% above the resting metabolic rate (ie, PAL >1.60 METs 24 h( 1)) may be necessary to maintain body weight in middle-age and can be achieved by incorporating 45-60 min of brisk walking, gardening/yardwork, or cycling into the daily routine. PMID- 15543160 TI - Food selection and eating behaviour during weight maintenance intervention and 2 y follow-up in obese men. AB - OBJECTIVE: The aim was to assess long-term changes in food consumption and eating behaviour during and 2 y after dietary counselling in weight-reduced obese men. DESIGN: Observational study from a randomised controlled trial. SETTING: Outpatient clinic of a research institute. SUBJECTS: A total of 36 subjects with complete data on food intake during the study. Subjects were obese (mean body mass index (BMI) 32.8 kg/m2) men aged 35-50 y, recruited by media advertising. INTERVENTIONS: Dietary counselling was included in 2 months weight reduction with very-low-energy-diet and in 6 months weight maintenance programme, which also included physical activity counselling. This was followed by a 23 months unsupervised follow-up with yearly assessments. Food intake was assessed six times during the study by 4-day food records. Eating behaviour was assessed by Three-Factor Eating Questionnaire (TFEQ). RESULTS: Increased consumption of low fat cheese, low-fat margarine, vegetables and high-fibre bread, and decreased consumption of sugar, sausage, high-fat cheese, high-fat margarine, fat products and sweets were observed during dietary counselling. Most of these changes returned later to prestudy consumption level. The relapse in dietary changes was partly associated with scoring low in restraint and high in disinhibition and hunger. CONCLUSION: In obese men, long-term maintenance of dietary changes was difficult. New ways to ease self-monitoring and increase self-efficacy might be necessary to improve maintenance of dietary changes. PMID- 15543161 TI - Reduction of risk factors for cardiovascular diseases in morbid-obese patients following biliary-intestinal bypass: 3 years' follow-up. AB - BACKGROUND: Obese patients are often affected by hypertension, dyslipidaemia, impaired glucose metabolism, and suffer from cardiovascular disease (CVD), related to the characteristic metabolic alterations. AIM OF THE STUDY: To evaluate reduction of risk factors for CVDs in morbid-obese patients (body mass index (BMI)>40 kg/m2) after weight loss upon bariatric surgery intervention of biliary-intestinal bypass. SUBJECTS: 45 (17 men, 28 women) morbid-obese patients (age: 19-49 y, BMI>40 kg/m2). All patients were selected on the basis of medical history, physical and biochemical evaluation and of psychiatric tests, which were performed on all individuals admitted to our Day Hospital to verify the safety of surgical intervention. MEASUREMENTS: Body weight, body composition (by dual X-ray absorptiometry, DXA), blood pressure, lipid profile, fibrinogen and glucose metabolism were monitored at baseline and 1, 3, 6, 9, 12, 24 and 36 months after surgery. RESULTS: A significant and persistent weight loss was present in all patients at the end of the 3 y follow-up period (P<0.001), with a progressive reduction of total and trunk fat mass as evaluated by means of DXA. Additionally, a parallel significant reduction in systolic (P<0.001) and diastolic (P<0.001) blood pressure was observed. Total and LDL cholesterol were significantly reduced (P<0.001), while HDL showed no modifications; triglycerides declined progressively during the 3 y follow-up (P<0.001). Fibrinogen decreased from 364.5+/-82.4 to 266.4+/-45.7 mg/dl at the end of the period (P<0.001). Fasting glucose levels and glucose levels 120 min after an oral glucose tolerance test were reduced from 95.1+/-20.3 to 78.6+/-9.1 mg/dl (P<0.001) and from 116.9+/-34.7 to 77.6+/-15.5 mg/dl (P<0.001), respectively, at baseline and at the end of the study. Moreover, fasting insulin decreased from 30.0+/-20.4 to 8.6+/-2.9 microUI/ml (P<0.001) after 3 y, while insulin levels after (120 min) oral glucose load decreased from 105.5+/-61.5 to 12.0+/-6.0 microUI/ml (P<0.001). CONCLUSION: Our results show that biliary-intestinal bypass may represent a valid and alternative therapeutic approach in patients with morbid obesity since it induces a significant and stable reduction of body weight and obesity-related risk factors for CVD. PMID- 15543163 TI - The Caenorhabditis elegans CED-9 protein does not directly inhibit the caspase CED-3, in vitro nor in yeast. AB - A genetically defined pathway orchestrates the removal of 131 of the 1090 somatic cells generated during the development of the hermaphrodite nematode Caenorhabditis elegans. Regulation of apoptosis is highly evolutionarily conserved and the nematode cell death pathway is a valuable model for studying mammalian apoptotic pathways, the dysregulation of which can contribute to numerous diseases. The nematode caspase CED-3 is ultimately responsible for the destruction of worm cells in response to apoptotic signals, but it must first be activated by CED-4. CED-9 inhibits programmed cell death and considerable data have demonstrated that CED-9 can directly bind and inhibit CED-4. However, it has been suggested that CED-9 may also directly inhibit CED-3. In this study, we used a yeast-based system and biochemical approaches to explore this second potential mechanism of action. While we confirmed the ability of CED-9 to inhibit CED-4, our data argue that CED-9 can not directly inhibit CED-3. PMID- 15543164 TI - Nuclear localization of FADD protein. PMID- 15543169 TI - Cytomegalovirus in aetiology of Posner-Schlossman syndrome: evidence from quantitative polymerase chain reaction. PMID- 15543168 TI - Iatrogenic eccentric full thickness macular holes following vitrectomy with ILM peeling for idiopathic macular holes. AB - PURPOSE: To document a previously unreported complication after vitrectomy with peeling of the internal limiting membrane (ILM) for idiopathic macular holes. METHOD: Retrospective review of notes of 232 consecutive patients who underwent vitrectomy with peeling of the ILM for idiopathic macular holes from 1996 to 2001. Four patients were found to have eccentric iatrogenic macular holes postoperatively. Optical coherence tomography was used to evaluate these holes. RESULTS: The idiopathic macular holes were graded from stages II to IV preoperatively with visual acuities from 6/18 to 6/60. All patients had surgery within 6 months of presentation. They underwent vitrectomy with complete separation of the posterior cortical vitreous, peeling of the ILM, injection of platelets (0.1 ml), and gas tamponade with SF6 20%. Postoperatively the patients postured strictly face down for 10 days. Follow-up ranged from 8 months to 6 years. Iatrogenic eccentric macular holes were noted postoperatively. The holes were located between 3 and 6 o'clock in three patients and at 9 o'clock in the fourth patient, relative to the macula. Optical coherence tomography showed them to be full thickness and completely flat. No further intervention was necessary. No complications have arisen during follow-up. COMMENT: To our knowledge iatrogenic eccentric full thickness macular holes after macular hole surgery have never been reported. We believe that the location of the holes represents the initial site of ILM elevation. These holes are asymptomatic, have not required any treatment and have not caused any complications in up to 6 years of follow up. PMID- 15543170 TI - Spontaneous hyphaema from pupillary vascular tufts in a patient with branch retinal vein occlusion. PMID- 15543171 TI - Intraocular pressure during haemodialysis: a review. AB - OBJECTIVES: Although several works in the past have examined the effect of haemodialysis (HD) on intraocular pressure (IOP), reported findings, theories, and conclusions are very different. The objectives of this article are to resume the reported evidence of IOP changes during HD, to review the proposed hypothesis of HD influence on IOP, and to determine if ophthalmic examination is imperative in HD patients. METHODS: We analysed the peer-reviewed English literature and selected all possible relevant articles. RESULTS: The influence of HD on IOP is not clear, and even in recent studies opposite findings can be found. CONCLUSIONS: Future studies are needed to clarify the effects of HD on IOP. In patients with glaucoma or with predisposed narrow angles, or eyes with impaired aqueous outflow, the possibility of acute IOP rise during HD could be much more frequent than in normal patients. So in these patients, a more strict ophthalmic scheduled examination seems to be feasible. PMID- 15543172 TI - The first cut is the deepest: basic surgical training in ophthalmology. AB - PURPOSE: To examine the basic surgical training received by Senior House Officers (SHOs) in ophthalmology and the influence on training of sociodemographic and organisational factors. METHODS: Cross-sectional survey of SHOs in recognised UK surgical training posts asking about laboratory training and facilities, surgical experience, demographic details, with the opportunity to add comments. RESULTS: A total of 314/466 (67%) questionnaires were returned. In all, 67% had attended a basic surgical course, 40% had access to wet labs and 39% had spent time in a wet lab in the previous 6 months. The mean number of part phakoemulsification (phako) procedures performed per week was 0.79; the mean number of full phakos performed per week was 0.74. The number of part phakos performed was negatively correlated, and the number of full phakos completed was positively correlated, with length of time as an SHO. Respondents who had larger operating lists performed more full phakos per week (P<0.001). Compared to men, women were less likely to have access to a wet lab (P=0.013), had completed fewer full phakos per week (P=0.003), and were less likely to have completed 50 full phakos (P=0003). SHOs' comments revealed concerns about their limited 'hands on' experience. CONCLUSIONS: There are significant shortcomings in the basic surgical training SHOs receive, particularly in relation to wet lab experience and opportunities to perform full intraocular procedures. SHOs themselves perceive their training as inadequate. Women are disadvantaged in both laboratory and patient-based training, but minority ethnic groups and those who qualified overseas are not. PMID- 15543173 TI - Ocular manifestations of head injury: a clinical study. AB - AIM: This prospective study aimed to clinically correlate the various ocular findings with the neurological status in cases of closed head injury. METHODS: A total of 200 consecutive cases of closed head injury admitted to a major teaching hospital underwent a thorough ophthalmic assessment. The Glasgow coma scale (GCS) and the Revised trauma score (RTS) were applied to grade the severity of injury and assess the prognosis. Kendall's tau-b and Fisher's exact test were used in the analysis. RESULTS: The main causes of head injury were road traffic accidents 52.5% followed by assaults in 34%. Ocular involvement was found in 167(83.5%) cases. These included corneal and scleral tears in 2%, subconjunctival haemorrhage or ecchymosis in 46%, orbital fractures 12%, pupillary involvement 6.5%, papilloedema 5.5%, intraocular trauma 5.5%, proptosis 3%, lateral rectus palsy 2%, lacrimal gland prolapse 1%, and optic nerve trauma 0.5%. All 21 patients (10.5%) who died had eye involvement. In all, 150 cases (75%) with a RTS of 12 had a good prognosis. Of these 124 (82.6%) had ocular involvement of no neurological significance. CONCLUSIONS: Although sophisticated imaging techniques are available to localize lesions, early ophthalmic assessment in correlation with the GCS aids in prognosticating outcomes. Pupillary involvement, papilloedema, and ocular motor paresis pointed to a more severe head injury. To our knowledge, this is the only prospective study recording ocular findings in the first few hours and attempting a correlation with the final outcome. PMID- 15543174 TI - One- and two-step hydrogen peroxide contact lens disinfection solutions against Acanthamoeba: how effective are they? AB - PURPOSE: Effective contact lens disinfection solutions are important to keep the storage case free of acanthamoebae and thus prevent an infection of the eye. The aim of our study was to evaluate the effectivity of two new one-step hydrogen peroxide disinfecting solutions against Acanthamoeba spp. and compare it to the effectivity of other commercially available systems. METHODS: Nine one-step 3% hydrogen peroxide systems including the new systems Silver Sept (platinum and silver disk for intensifying disinfection) and Blue Vision (newly composed catalytic tablet) and 2 two-step systems (0.6 and 3.0% H(2)O(2)) were tested for their effectivity against cysts of two Acanthamoeba keratitis isolates at different concentrations. RESULTS: After a soaking time of 8 h (overnight soaking of contact lenses) the 2 two-step systems completely destroyed the cysts of both Acanthamoeba strains, even at the highest concentration of cysts tested. The nine tested one-step systems showed weaker effects. The new Blue Vision system was able to eradicate the cysts of one strain at the low concentration of cysts. CONCLUSIONS: One-step hydrogen peroxide systems do not have sufficient effects on Acanthamoeba cysts and therefore may not protect the contact lens user from a possible infection of the eye. Further development of tablets like the ones used in the Blue Vision system may result in better cysticidal effects for one-step systems. PMID- 15543175 TI - The impact of national diabetic retinopathy screening on ophthalmology: the need for urgent planning. PMID- 15543176 TI - Hemiretinal arterial supply by the cilioretinal artery. PMID- 15543177 TI - Acute nonarteritic ischaemic optic neuropathy after phentermine. PMID- 15543178 TI - Reverse self-sealing sclerostomies. PMID- 15543179 TI - New paradigms in the mechanisms and management of glaucoma. AB - During the last 30 years, the definition of glaucoma as been revised to eliminate the inclusion of intraocular pressure. Open angle glaucoma is the second leading cause of blindness in the world, but the proportion of those with the disease who become blind is low. Diagnostic methods for glaucoma need improvement. The pathogenetic steps to loss of neurons in glaucoma are increasingly understood and non-pressure lowering therapies are on the horizon. PMID- 15543180 TI - Kinetic anaesthesia for laser surgery. AB - PURPOSE: Local anaesthetic is widely used in ophthalmic surgery and more recently in vitreoretinal surgery. Akinesia is a useful effect of local anaesthetic blocks, but there are situations where some residual globe movements are of benefit. We looked to see whether reducing the volume of anaesthetic solution used in a block could retain some kinesia while achieving good analgesia. METHODS: We compared two groups of patients undergoing panretinal photocoagulation (PRP) with an indirect laser. The control group received 5 ml of anaesthetic solution in a single injection by a standard intraconal technique; a second group received a lower volume of solution by the same technique. We recorded the adequacy of anaesthesia and the amount of residual kinesia for the two groups. Differences between groups were analysed using the Student's t-test and chi(2) tests. RESULTS: The low-volume group received an average of 2.8 ml, compared to 5 ml in the control group. There was no significant difference in the adequacy of analgesia achieved, however 16/18 (89%: 95% confidence intervals (CI)=81.5-96.3%) of the low-volume group had good perioperative kinesia compared to just 3/21 (14%: 95% CI=6.6-21.9%) of the controls (P<0.001). CONCLUSIONS: We have shown that low-volume intraconal blocks retain some perioperative kinesia without compromising their analgesic effect. PMID- 15543181 TI - Radiotherapy for age-related macular degeneration: no more pilot studies please. PMID- 15543182 TI - Results of radiotherapy of subfoveal neovascularization with 16 and 20 Gy. AB - PURPOSE: In a nonrandomized, prospective study the efficacy of radiotherapy with 16 and 20 Gray (Gy) for subfoveal neovascularization in age-related macular degeneration (ARMD) was analysed. MATERIAL AND METHODS: From 1996 to 1998, 63 eyes were irradiated with 16 Gy and 38 eyes with 20 Gy for exudative ARMD. A total of 12 eyes had classic ARMD, 89 eyes occult ARMD, median baseline visual acuity (VA) was 6/30 (range: 3/60-6/9.5), median age was 78 years. Risk factors (type of ARMD, baseline VA) were evenly distributed in both groups. Median follow up was 1.3 years (range: 4 months-4.7 years). VA of +/-1 line or better and unchanged size and activity of the membrane in fluorescein angiography were defined as stable. Actuarial methods were used. RESULTS: Median loss of VA was -3 lines (range: -14 to +5), neovascularization remained unchanged or decreased in size and activity in 35 eyes. At 18 months, the probability of stabilized VA was 0.4 (95% confidence interval (CI): 0.3-0.5), at 24 months 0.3 (95% CI: 0.2-0.4). Radiation dose, type of ARMD or baseline VA had no significant impact on outcome of VA and membrane size and activity (P>0.05). Side effects were mild and transient increased tearing. CONCLUSION: In this study, the results after radiotherapy were comparable to the natural course of the disease. An impact of radiation dose (16 vs 20 Gy) on stabilizing visual acuity and subfoveal neovascularization could not be shown. The results of studies on dose escalation using very small fields and high radiation doses should be awaited. PMID- 15543183 TI - Anterior capsular phimosis with complete occlusion of the capsulorhexis opening. PMID- 15543184 TI - Endophthalmitis following 25-gauge vitrectomy. PMID- 15543186 TI - The proview phosphene tonometer: a clinical evaluation. PMID- 15543187 TI - The influence of pharmacological mydriasis on biomicroscopic evaluation of the glaucomatous optic nerve head. AB - AIMS: To investigate whether pharmacological mydriasis influences interobserver agreement or within-observer agreement (comparing estimates made before and after dilation) in the evaluation of the optic nerve head (ONH) of the glaucoma suspect or patient. METHODS: Monoscopic disc photographs of the ONH were assessed by each observer on two separate occasions in order to establish baseline intra- and inter-observer agreement. Then the ONH of 53 eyes of 53 patients was examined by each observer on two separate occasions, the pupil being pharmacologically dilated on only one of these visits. Each observer commented on the vertical and horizontal cup-to-disc (C/D) ratio, and the presence of the following ONH parameters: laminar dots; disc haemorrhage; disc saucering; disc notching, and peripapillary atrophy (PPA). RESULTS: Intersessional variability of C/D ratio estimates, based on photographs of the ONH, was similar for the two observers. The mean (+/-standard deviation) age of the 53 patients was 70 (+/-15) years, and the male : female ratio was 28 : 25. Intraobserver agreement of C/D ratio estimation performed through a dilated pupil on one occasion and an undilated pupil on the other occasion was not statistically different between observers. Interobserver agreement of C/D ratio estimates were not adversely affected in a statistically meaningful way if ONH evaluation was performed by each observer under conditions of nonmydriasis or by each observer under conditions of pharmacological mydriasis. Within-observer (before and after dilation) and interobserver agreement with respect to the presence of laminar dots (kappa=0.41 0.69), PPA (kappa=0.36-0.54), and pathological cupping (kappa=0.46-0.68) was typically moderate to good across the two visits where the pupil was dilated on one occasion only, but less reliable for saucering, disc notching and disc margin haemorrhage. CONCLUSION: Routine pharmacological mydriasis is not essential for reproducible evaluation of the ONH for patients in whom a satisfactory view of the optic disc can be achieved through the undilated pupil. PMID- 15543188 TI - Smoking delays the response to treatment in episcleritis and scleritis. AB - PURPOSE: To evaluate the influence of smoking on comorbidity, treatment, visual and general outcome in patients with scleritis. METHODS: The smoking habits of 103 patients with a diagnosis of episcleritis or scleritis were evaluated. These patients were treated by one ruling protocol at the Leiden University Medical Center between 1997 and 2000. Medical records of each patient were evaluated in detail. Data on possible factors concerning smoking were collected by postal questionnaire. RESULTS: Of all 103 patients diagnosed with either episcleritis or scleritis, 41 (39.8%) were smoking during treatment of the scleral inflammation. In total, 19 patients (18.4%) had a smoking history while 43 (41.7%) patients have never smoked. The response to any of the given medications could be delayed by at least 4 weeks in many smoking patients (odds ratio (OR) 5.4 [95% confidence interval 1.9-15.5]), particularly those with posterior scleritis. Smoking patients above the age of 48 years were even more likely to respond belatedly to any given therapy (OR 6.6 [2.1-20.7]). However, having a smoking history did not delay the response. Furthermore, smoking did not worsen the visual prognosis and was not associated with additional recurrences or ocular complications after successful treatment. CONCLUSIONS: Although scleritis patients who smoked during treatment eventually responded, there was frequently over a month's delay before the medication became effective when compared to nonsmokers. This was irrespective of the type of disease or given therapy. As a consequence, smokers required more intensive therapy than those who did not smoke. PMID- 15543189 TI - Argon laser photocoagulation for retinopathy of prematurity: long-term outcome. AB - PURPOSE: To assess the visual, refractive, ocular structural, and neurodevelopmental outcome in a group of children who were screened, treated with laser photocoagulation for threshold retinopathy of prematurity (ROP), and followed up at our institution. METHODS: The 21 survivors of a cohort of 23 children were recalled at the age of 5 years or more. They underwent a full ophthalmologic examination. A paediatric neurologist and/or developmental paediatrician performed a neurodevelopmental assessment and a neuropsychologist performed psychological testing. RESULTS: A total of 30 eyes (71.4%) had a best corrected visual acuity (BCVA) of 20/40 or better. In all, 26 (62%) eyes were myopic; the overall mean spherical equivalent was -4.95 D. No eye had Stage 4 findings and only one eye (2.4%) progressed to stage 5 despite photocoagulation. In total, 14 patients (66.6%) showed evidence of stereopsis. Strabismus was seen in six patients (28.5%) and nystagmus in three (14.3%). Periventricular leukomalacia (PVL) was identified in five patients (23.8%), cerebral palsy (CP) in seven (33.3%), and intraventricular haemorrhage (IVH) in 13 (62%). Cerebral palsy and IVH associated strongly with visual acuity of less than 20/40 (P=0.009 and 0.047). There was also a strong association between visual acuity of 20/40 or above and a better cognitive outcome (P=0.013). CONCLUSIONS: Retinal scarring and detachment are relatively rare causes of visual morbidity 5 years or more after laser treatment for threshold ROP, but the neurological sequelae of extreme prematurity and amblyopia remain important causes of impaired visual function. PMID- 15543191 TI - Bilateral vitreous haemorrhage associated with dengue fever. PMID- 15543190 TI - Outcome of pterygium surgery: analysis over 14 years. AB - AIM: To report the outcome of pterygium surgery performed at a tertiary eye care centre in South India. METHODS: Retrospective analysis of medical records of 920 patients (989 eyes) with primary and recurrent pterygia operated between January 1988 and December 2001. The demographic variables, surgical technique (bare sclera, primary closure, amniotic membrane transplantation (AMT), conjunctival autograft (CAG), conjunctival-limbal autograft (CLAG), or surgical adjuvants), recurrences and postoperative complications were analysed. RESULTS: A total of 496 (53.9%) were male and 69 (7.5%) had bilateral pterygia. Bare sclera technique was performed in 267 (27.0%) eyes, primary conjunctival closure in 32 (3.2%), AMG in 123 (12.4%), CAG in 429 (43.4%), and CLAG in 70 (7.1%). Adjuvant mitomycin C was used in 44 (4.4%) cases. The mean duration of follow-up was 8.9+/-17.0 and 5.9+/-8.8 months for unilateral primary and recurrent pterygia, respectively. The overall recurrence rate was 178 (18.0%). Following primary and recurrent unilateral pterygium excision respectively, recurrences were noted in 46 (19.4%) and 1 (33.3%) eyes after bare sclera technique, five (16.7%) and 0 after primary closure, 28 (26.7%) and 0 with AMG, 42 (12.2%) and five (31.3%) with CAG, and nine (17.3%) and two (40%) with CLAG. Recurrences were significantly more in males with primary (23.3 vs 10.7%, P<0.0001) and recurrent (26.7 vs 0%, P=0.034) pterygia, and in those below 40 years (25.2 vs 14.8%, P=0.003). CONCLUSION: CAG appears to be an effective modality for primary and recurrent pterygia. Males and patients below 40 years face greater risk of recurrence. Bare sclera technique has an unacceptably high recurrence. Prospective studies comparing CAG, CLAG, and AMG for primary and recurrent pterygia are needed. PMID- 15543192 TI - Keratoacanthoma of the lower eyelid. PMID- 15543193 TI - Phacoemulsification and intraocular lens implantation following pars plana vitrectomy: a prospective study. PMID- 15543194 TI - Cardiac toxicity of high-dose chemotherapy. AB - Cardiac toxicity is an uncommon but potentially serious complication of high-dose (HD) chemotherapy and little is known about incidence, severity and underlying mechanisms. We have systematically reviewed the literature of the last 30 years to summarize and appraise the published evidence on cardiac toxicity associated with HD chemotherapy. HD cyclophosphamide-containing regimens have been most commonly associated with cardiac toxicity, with a progressively decreasing incidence over time. Dosage, application regimens and coadministration of other chemotherapeutic agents emerged as risk factors. While cardiac toxicity has been rarely associated with other cytotoxic drugs, an unexpected incidence of severe cardiotoxicity resulted from reduced-intensity conditioning regimens containing melphalan and fludarabine. Predictive value of cardiologic examination of patients is limited, and patients with a slight depression of cardiac performance could tolerate HD chemotherapy. Clinical examination, resting electrocardiography and dosage adjustment in overweight patients remain the mainstay of prevention, with bidimensional echocardiography (2D echo) for patients with a history of anthracycline exposure. Strategies to decrease the long-term negative impact of anthracycline administration on cardiac performance are being investigated. New 2D echo-based techniques and circulating markers of cardiac function hold promise for allowing identification of patients at high risk for and early diagnosis of cardiac toxicity. PMID- 15543195 TI - Long-term follow-up of nonmyeloablative allogeneic stem cell transplantation for renal cell carcinoma: The University of Chicago Experience. AB - Nonmyeloablative allogeneic stem cell transplantation (NST) has considerable activity in patients with metastatic renal cell carcinoma (RCC), although there are limited long-term follow-up data. Between February 1999 and May 2003, 18 patients with metastatic RCC underwent 19 matched-sibling NSTs after conditioning with fludarabine and cyclophosphamide with tacrolimus and mycophenolate mofetil as post-transplant immunosuppression. Among the four objective responses, all were partial and have relapsed with a median response duration of 609 days (range, 107-926). All responders are alive at a median of 41 months. Median overall survival for the entire cohort was 14 months. There were four early treatment-related deaths and one late treatment-related death. Eight patients died from progressive disease and five (28%) from treatment-related mortality. Stratifying transplant outcome as early death, intermediate (no response, no early death), or response, the combination of pre-treatment anemia and decreased performance status, was associated with adverse outcome (P = 0.015) and reduced survival (HR 5.4, 95% confidence interval of 1.4 to 21, P = 0.007). Responders demonstrated prolonged survival compared to nonresponders (P = 0.002). NST leads to durable responses in a minority of metastatic RCC patients. Appropriate patient selection is paramount. Anemia and decreased performance status may enable risk stratification. PMID- 15543196 TI - Cure of myeloma: hype or reality? AB - High-dose treatment (HDT) with autologous stem cell transplant(s) (ASCT) improved survival, when compared to standard treatment, in multiple myeloma patients. Although the superiority of HDT is clearly recognized by the medical community, what is less appreciated is the disproportionate benefit enjoyed (as a result of this approach) by various patient subgroups. As the clinical heterogeneity of myeloma can be currently traced to its underlying genetic features, prognostically different patient groups can be identified largely based on the presence of adverse cytogenetic abnormalities and high serum levels of lactate dehydrogenase at baseline (high-risk features). While HDT applied to high-risk patients leads to modest survival gains, the same treatment, as the backbone of a comprehensive approach, can be curative in a minority of low-risk patients. A third group of low-risk patients will enjoy rather prolonged (10-year) survival, interrupted, however, by responsive relapses. In a manner analogous to follicular lymphoma, this latter group may transform to a more aggressive disease, characterized by the new acquisition of adverse cytogenetic abnormalities. Improving the complete response rate in these patients, by integrating newer therapeutic agents, may increase their cure rate. Currently non-myeloablative, allogeneic transplants (and possibly proteasome inhibitors) are the most promising approaches for high-risk patients. PMID- 15543197 TI - Stem cell factor improves SCID-repopulating activity of human umbilical cord blood-derived hematopoietic stem/progenitor cells in xenotransplanted NOD/SCID mouse model. AB - Poor in vivo homing capacity of hematopoietic stem/progenitor cells (HS/PCs) from umbilical cord blood (UCB) can be reversed by short-term ex vivo manipulation with recombinant human stem cell factor (rHuSCF). This study was designed to evaluate the effect of ex vivo manipulation of UCB-derived HS/PCs with rHuSCF on human cell engraftment rates in xenotransplanted NOD/SCID mouse model. The human cell engraftment rates in xenotransplanted primary and secondary NOD/SCID mice were characterized using four-color flow cytometric analysis and progenitor assay. Grafts of rHuSCF-treated UCB CD34(+) cells resulted in significantly higher levels of human cell engraftment than that of nontreated ones in both xenotransplanted primary and secondary NOD/SCID recipients. Fresh UCB CD34(+) cells did not express either of the matrix metalloproteinase (MMP) family members MMP-2 or MMP-9. rHuSCF-treated UCB CD34(+) cells expressed significant levels of MMP-2 and MMP-9. Pretreatment of UCB CD34(+) cells with the specific MMP inhibitor completely blocked human cell engraftment in xenotransplanted NOD/SCID recipients. Our results indicate that ex vivo manipulation of human HS/PCs with rHuSCF might provide an optimal approach to develop more effective stem cell based therapies in situations where engraftment is delayed due to limiting HS/PCs number, for example, UCB transplantation. PMID- 15543198 TI - Successful haemopoietic stem cell transplantation does not correct mannan-binding lectin deficiency. AB - It has been reported that in allogeneic stem cell transplantation, the mannan binding lectin (MBL) status of the donor has prognostic value for the recipient. Two MBL-deficient patients, with coexisting haematological malignancy, were identified who were treated with bone marrow from donors with normal MBL concentrations. Although both patients engrafted successfully and remain in complete remission, neither seroconverted to the MBL sufficiency status of his donor over a follow-up period exceeding 2 years. This does not support the concept of MBL replacement by stem cell therapy, and does not provide an explanation for high MBL concentrations in stem cell donors protecting recipients from post transplant infections. PMID- 15543199 TI - T-cell depletion improves outcome after autologous stem cell transplant in patients with systemic lupus erythematosus (SLE). PMID- 15543200 TI - Prognostic factors for survival after autologous transplantation: a single centre experience in 133 multiple myeloma patients. AB - Autologous stem cell transplantation (ASCT) has an established role in the treatment of symptomatic multiple myeloma (MM). Our aim was to analyse the impact of selected prognostic parameters on the survival of patients with MM after ASCT. The new International Staging System (ISS) was also evaluated. A total of 133 MM patients were transplanted in our centre between 1995 and 2002. Following ASCT, 35% of patients were in complete remission (CR) and 60% were in partial remission (PR). The median progression-free (PFS) and overall (OS) survival from transplantation were 29.5 and 68.8 months, respectively. Transplant-related mortality (TRM) was 3%. On multivariate analysis, factors associated with significantly shorter OS were lack of CR after transplant (P = 0.002, hazard ratio (HR): 3.1), stage 3 according to ISS (P = 0.001, HR: 3.0) and age at transplant over 60 years (P = 0.035, HR: 2.0). The status of disease before ASCT did not significantly affect PFS and OS after transplantation. We conclude that ASCT is a safe and effective procedure in MM patients, associated with low TRM. The survival after ASCT was dependent on response after ASCT, stage according to ISS and age. PMID- 15543201 TI - Use of intrathecal prophylaxis in allogeneic haematopoietic stem cell transplantation for malignant blood diseases: a survey of the European Group for Blood and Marrow Transplantation (EBMT). AB - SUMMARY: A survey was carried out among EBMT centres to describe the current practice concerning intrathecal (i.t.) prophylaxis in allogeneic stem cell transplantation for malignant diseases in patients with no central nervous system (CNS) manifestations of the disease at any time. A total of 90 centres reported their practice: 42 centres (47%) never used pre-transplant i.t. prophylaxis as part of the conditioning, whereas 48 centres (53%) gave i.t. prophylaxis to selected groups. The main indications were acute lymphoblastic leukaemia (ALL), acute myeloid leukaemia (AML), and lymphoma (53, 33, and 23% of all centres, respectively). Prophylaxis was usually given to all patients with ALL, but often restricted to high-risk patients in AML and lymphoma. Of the 90 centres, 29 (32%) gave prophylactic i.t. treatment after the transplantation, mainly for the same indications as pre-transplant. This survey illustrates the heterogeneity in the current practice of i.t. prophylaxis in allogeneic transplantation for malignant blood disorders in Europe. The documentation in the literature to support the use of i.t. prophylaxis as part of transplantation for malignant diseases in patients without preceding CNS involvement is sparse. Based on the rarity of isolated CNS relapse after allogeneic stem cell transplantation, EBMT does not recommend routine i.t. prophylaxis to patients without prior CNS involvement. PMID- 15543202 TI - In vivo characterization of developing chronic pancreatitis in rats. AB - Despite numerous experimental and clinical investigations, there is no unifying concept on pathophysiology and pathogenesis of chronic pancreatitis. Defining the interplay between pancreatic microcirculation and parenchymal tissue, we will provide a basis for the better understanding of pancreatic fibrogenesis using in vivo high-resolution multifluorescence microscopy in dibutyltin chloride (DBTC) exposed rats. Pancreatic microcirculation at days 3 and 7 after DBTC revealed leukocyte activation with a two-fold higher fraction of rolling cells and a nine- to 10-fold increase of cells firmly adherent to the endothelial lining, followed by subsequent transendothelial migration into tissue, as given by chloracetate esterase histology. In vivo staining of acinar tissue with bisbenzimide presented single cells exhibiting nuclear chromatin condensation and fragmentation. Apoptotic cell death was confirmed by immunohistochemical staining for active caspase-3 as well as by TUNEL analysis. Necrotic cells were found dispersed throughout the exocrine tissue under observation. Both modes of cell death were found highest in extent at days 3 and 7 with 15-20 cells/mm2, but progressively decreased below 10 cells/mm2 up to 28 days after DBTC. By means of in vivo microscopy yellow-green autofluorescent collagen deposits were found at day 7 and progressively increased up to approximately 12% at day 28 after DBTC. Concomitantly, density of capillaries progressively decreased and capillaries failing to conduct blood flow became apparent. Present on-line analysis indicates an early inflammatory response with acinar cell death, most probably triggering progression of disease with collagen deposition, capillary rarefication and manifestation of perfusion failure. These temporal and spatial multiparameter measurements of the in vivo microenvironment provide new insights into the pathological processes of pancreatic fibrogenesis. PMID- 15543203 TI - Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes. AB - For interpretation of quantitative gene expression measurements in clinical tumor samples, a normalizer is necessary to correct expression data for differences in cellular input, RNA quality, and RT efficiency between samples. In many studies, a single housekeeping gene is used for normalization. However, no unequivocal single reference gene (with proven invariable expression between cells) has been identified yet. As the best alternative, the mean expression of multiple housekeeping genes can be used for normalization. In this study, no attempt was made to determine the gold-standard gene for normalization, but to identify the best single housekeeping gene that could accurately replace the measurement of multiple genes. Expression patterns of 13 frequently used housekeeping genes were determined in 80 normal and tumor samples from colorectal, breast, prostate, skin, and bladder tissues with real-time quantitative RT-PCR. These genes included, large ribosomal protein, beta-actin, cyclophilin A, glyceraldehyde-3 phosphate dehydrogenase, phosphoglycerokinase 1, beta-2-microglobin, beta glucuronidase, hypoxanthine ribosyltransferase (HPRT), TATA-box-binding protein, transferrin receptor, porphobilinogen deaminase, ATP synthase 6, and 18S ribosomal RNA. Principal component analysis was used to analyze these expression patterns, independent of the level of expression. Our approach identified HPRT as the single best reference gene that could be used as an accurate and economic alternative for the measurement of multiple housekeeping genes. We recommend this gene for future studies to standardize gene expression measurements in cancer research and tumor diagnostics until a definite gold standard has been determined. PMID- 15543204 TI - Overexpression of fatty acid synthase in chemically and hormonally induced hepatocarcinogenesis of the rat. AB - Fatty acid synthase (FAS) is the key enzyme of de novo fatty acid synthesis and has been shown to be involved in carcinogenesis of numerous human malignancies, including breast, colorectal, and prostate carcinomas, often associated with a worse prognosis. Although FAS is mainly expressed in the liver, an implication of FAS in hepatocarcinogenesis, has not yet been investigated. FAS expression is stimulated by insulin and glucose, and insulin is also the primary trigger of hepatocarcinogenesis in an endocrine experimental model, which is induced by low number transplantation of islets of Langerhans into the livers of diabetic rats. We therefore investigated, whether FAS is implicated in hepatocarcinogenesis in this model and in comparison to chemically induced hepatocarcinogenesis after N nitrosomorpholine (NNM) treatment in diabetic and normoglycemic rats. Preneoplastic clear-cell foci of altered hepatocytes (FAH), harvested after laser microdissection of kryostat sections, showed an overexpression of FAS messenger RNA in gene expression profiles, done by array-hybridization, and in quantitative RT-PCR (Light-Cycler). Virtually, all (96-98%) of the subsequently investigated FAH and the glycogenotic hepatocellular adenomas and carcinomas showed an additional strong FAS protein overexpression. In the NNM-model, FAS protein was also overexpressed in the vast majority (87%) of glycogenotic FAH and neoplasms, in particular in the diabetic animals. Also two spontaneous glycogenotic FAH in control animals displayed strong FAS overexpression. Basophilic lesions and neoplasms, which occasionally develop out of the primary clear-cell FAH at later stages of carcinogenesis, however, lost FAS overexpression. In conclusion, FAS overexpression is an early phenonemon in spontaneous, hormonally and chemically induced rat hepatocarcinogenesis, demonstrable in early clear-cell (glycogenotic) FAH and hepatocellular neoplasms. FAS overexpression can be attributed to the local hyperinsulinemia in the transplantation model and belongs to cellular and metabolic alterations in the chemical model, which are induced by an insulinomimetic but yet unknown mechanism. PMID- 15543205 TI - Potential role of heme oxygenase-1 in the progression of rat adjuvant arthritis. AB - Rat adjuvant arthritis is an experimental model widely used to evaluate etiopathogenetic mechanisms in chronic inflammation. We have examined the participation of heme oxygenase-1 (HO-1) in this experimental arthritis. In this study, an increased nitric oxide (NO) production in the paw preceded the upregulation of HO-1, whereas selective inhibition of inducible NO synthase (iNOS) after the onset of arthritis decreased HO-1 expression, suggesting that the induction of this enzyme may depend on NO produced by iNOS. Therapeutic administration of the HO-1 inhibitor tin protoporphyrin IX was able to control the symptoms of arthritis. This agent significantly decreased leukocyte infiltration, hyperplastic synovitis, erosion of articular cartilage and osteolysis, as well as the production of inflammatory mediators. In this experimental model, HO-1 can be involved in vascular endothelial growth factor production and angiogenesis. These results support a role for HO-1 in mediating the progression of the disease in this model of chronic arthritis. PMID- 15543206 TI - EGF-induced proliferation of adult human pancreatic duct cells is mediated by the MEK/ERK cascade. AB - Human postnatal pancreatic duct cells are a potential source of new beta cells. Factors regulating proliferation of human pancreatic duct cells in vitro are unknown. In several other cell types, this process is influenced by ligands of the ErbB receptor family. The expression and functionality of the ErbB family members and their possible role in duct cell proliferation were determined. In cultured adult human pancreatic duct cells the different members of the ErbB family (ErbB1-4) were present at transcript and protein level. Stimulation of the duct cells with epidermal growth factor (EGF) and betacellulin results in Tyr phosphorylation of ErbB1 and ErbB2, followed by activation of Shc, MEK1/2 and ERK1/2. Duct cells with activated ErbB signaling changed morphology and motility. EGF induced proliferation of a fraction of the duct cells and treatment with PD98059 prevented Ki67 expression in EGF-supplemented cells. When transduced with recombinant adenovirus expressing constitutively activated MEK1, duct cells proliferate and spread even in the absence of EGF. Importantly, the adult human duct cells retain their capacity to recapitulate ngn3-induced embryonic (neuro)endocrine differentiation after proliferation. Therefore, the present data support a possible role for human adult pancreatic duct cells, following expansion and transdifferentiation, as a source of insulin by transplantation to type I diabetes patients. PMID- 15543208 TI - Overweight and obesity epidemic among children. Answer from European countries. AB - OBJECTIVE: To investigate the methods and results used and conclusions found in available published papers on childhood overweight and obesity in Europe. SURVEYS: This paper compares the two available published papers on the prevalence of child and adolescent overweight and obesity in Europe. The first paper was published in November 2003 and was based on 20 previously conducted surveys performed from 1992 to 2001. The other paper was published in January 2004 and was based on data from the WHO collaborative survey 'Health Behaviour in School Children', which collected the data in 1997-1998 within a period of 9 months. Height and weight were included as optional questions and 13 European countries, Israel and United States participated. METHODS: In both papers, body mass index (BMI) was used. In the first paper, measured BMI was used and prevalence of overweight was calculated using IOTF cutoff points. In the other paper, overweight and obesity prevalences were calculated from self-reported height and weight using an internal study reference standard. RESULTS: The first paper found a north-south trend in overweight in Europe, whereas the second found a more equal distribution of overweight in European children. Thus, overweight was significantly increased among 13 y olds of both sexes in Finland, Ireland, and Greece, and in Portuguese girls. Among 15 y olds, the prevalence of overweight was significantly increased in Greek boys, and in Danish and Portuguese girls. On the contrary, in Lithuania, there was a significantly low prevalence of overweight among both ages and both sexes. CONCLUSION: Critical reviews of the results of the two published papers show that the year of data collection, methods and use of appropriate statistics are of critical importance for the conclusion drawn from comparative epidemiological surveys on the prevalence of overweight. PMID- 15543209 TI - Role of television in childhood obesity prevention. AB - OBJECTIVE: To assess the role of television as tool for childhood obesity prevention. METHOD: Review of the available literature about the relationship between television and childhood obesity, eating habits and body shape perception. RESULTS: The reviewed studies showed the following: television watching replaces more vigorous activities; there is a positive correlation between time spent watching television and being overweight or obese on populations of different age; obesity prevalence has increased as well as the number of hours that TV networks dedicate to children; during the last 30 y, the rate of children watching television for more than 4 h per day seems to have increased; children are exposed to a large number of important unhealthy stimulations in terms of food intake when watching television; over the last few years, the number of television food commercials targeting children have increased especially when it comes to junk food in all of its forms; the present use of food in movies, shows and cartoons may lead to a misconception of the notion of healthy nutrition and stimulate an excessive intake of poor nutritional food; and obese subjects shown in television programmes are in a much lower percentage than in real life and are depicted as being unattractive, unsuccessful and ridiculous or with other negative traits and this is likely to result in a worsening of the isolation in which obese subjects are often forced. The different European countries have different TV legislations. CONCLUSION: The usual depiction of food and obesity in television has many documented negative consequences on food habits and patterns. The different national regulations on programs and advertising directed to children could have a role in the different prevalence of childhood obesity in different European countries. Television could be a convenient tool to spread correct information on good nutrition and obesity prevention. PMID- 15543213 TI - Micro-environmental and socio-demographic determinants of childhood obesity. AB - AIM: To review the available data in Spain about the socio-demographic and home environmental determinants in children and adolescents. METHOD: Review of the main studies conducted in Spain that have analysed the relationship between overweight and socio-economic and environmental determinants in children and adolescents. RESULTS: In children aged 6-7 y from Aragon (Spain), the socio demographic determinants of childhood overweight were size of municipality, year of examination, gender, and province; in adolescents aged 13-14 y, the socio demographic determinants were year of examination, type of school, size of municipality, gender, and province; overweight showed a significant positive main effect with public schools and low municipality size. In a nationally representative sample of Spanish adolescents from 13 to 18.5 y (AVENA Study), there was a significant relationship between overweight and socio-economic status in males but not in females; in males, the lowest overweight prevalences were observed in both extreme socio-economic groups; moreover, overweight prevalences increased when socio-economic status decreased, from the high to the medium-low socio-economic group. The studied variables related with family environment did not show any significant effect in overweight prevalence. CONCLUSION: Better knowledge of the relationship between social class and childhood obesity would lead to clearer hypotheses for the relationship in adults and might improve the preventive measures by identifying children at risk. PMID- 15543214 TI - The nutrition transition: worldwide obesity dynamics and their determinants. AB - OBJECTIVE: This paper explores the major changes in diet and physical activity patterns around the world and focuses on shifts in obesity. DESIGN: Review of results focusing on large-scale surveys and nationally representative studies of diet, activity, and obesity among adults and children. SUBJECTS: Youth and adults from a range of countries around the world. MEASUREMENTS: The International Obesity Task Force guidelines for defining overweight and obesity are used for youth and the body mass index > or =25 kg/m(2) and 30 cutoffs are used, respectively, for adults. RESULTS: The nutrition transition patterns are examined from the time period termed the receding famine pattern to one dominated by nutrition-related noncommunicable diseases (NR-NCDs). The speed of dietary and activity pattern shifts is great, particularly in the developing world, resulting in major shifts in obesity on a worldwide basis. Data limitations force us to examine data on obesity trends in adults to provide a broader sense of changes in obesity over time, and then to examine the relatively fewer studies on youth. Specifically, this work provides a sense of change both in the United States, Europe, and the lower- and middle-income countries of Asia, Africa, the Middle East, and Latin America. CONCLUSION: The paper shows that changes are occurring at great speed and at earlier stages of the economic and social development of each country. The burden of obesity is shifting towards the poor. PMID- 15543215 TI - Epidemiology of childhood obesity--methodological aspects and guidelines: what is new? AB - INTRODUCTION: It is still a matter of debate as to how to define obesity in young people, although a growing consensus is to use body mass index (BMI) cutoffs to classify obesity in children and adolescents. OBJECTIVE: This article provides a brief overview of issues related to the assessment of obesity in children and adolescents. RESULTS: At present, BMI is probably the best choice among available measures. BMI can be easily assessed at low cost, and has a strong association with body fatness and health risks. However, as an indirect measure of adipose tissue, BMI has a number of limitations. Cole et al published a set of sex- and age-specific BMI cutoffs, which had been developed based on data collected in six countries, and the reference has been recommended for international use. Recently, several researchers have raised concerns regarding this international reference. It has been argued that population-specific standards should be used due to biological differences between populations. CONCLUSION: BMI is a valid and feasible indirect measure of body fatness, but it suffers from a number of limitations. More efforts are needed to develop valid classifications of childhood obesity. PMID- 15543216 TI - Genes, lifestyles and obesity. AB - OBJECTIVE: To review the role of genes and lifestyle factors, particularly dietary habits and physical activity patterns, in obesity risk as well as their potential interactions. DESIGN AND MEASUREMENTS: A descriptive report of a number of genes definitely ascribed or potentially implicated in excessive fat accumulation leading to obesity as assessed by different research approaches (Mendelian transmission, genetic animal models, epidemiological association/linkage studies and genome-wide scans). Also, the involvement of macronutrient intake and composition (fat/carbohydrate) as well as the role of activity-linked energy expenditure in obesity onset is reviewed. RESULTS: Examples of the role of the genotype as well as of the dietary macronutrient composition/intake and sedentary/low energy cost of physical activities in obesity prevalence are reported. CONCLUSIONS: Both genes and everyday life environmental factors such as cultural and social mediated food intake and reduced domestic and living work activities are involved in the obesity pandemia. The occurrence of gene x gene and gene x environmental factors interactions makes it more difficult to interpret the specific roles of genetics and lifestyle in obesity risk. PMID- 15543217 TI - Gene-gene interaction between PPAR gamma 2 and ADR beta 3 increases obesity risk in children and adolescents. AB - AIMS: Multiple genes are likely to be involved in obesity and these genes may interact with environmental factors to influence obesity risk. Our aim was to explore the synergistic contribution of the two polymorphisms: Pro12Ala of the PPAR gamma 2 gene and Trp64Arg of the ADR beta 3 gene to obesity risk in a Spanish children and adolescent population. METHODS: We designed a sex- and age matched case-control study. Participants were 185 obese and 185 control children (aged 5-18 y) from the Navarra region, recruited through Departments of Pediatrics (Hospital Virgen del Camino, Navarra University Clinic and several Primary Health Centers). The obesity criterion (case definition) was BMI above the 97th percentile according to Spanish BMI reference data for age and gender. Anthropometric parameters were measured by standard protocols. The genotype was assessed by PCR-RFLP after digestion with BstUI for PPAR gamma 2 mutation and BstNI for ADR beta 3 variants. Face-to-face interviews were conducted to assess the physical activity. Using a validated physical activity questionnaire, we computed an activity metabolic equivalent index (METs h/week), which represents the physical exercise during the week for each participant. Statistical analysis was performed by conditional logistic regression, taking into account the matching between cases and controls. RESULTS: Carriers of the polymorphism Pro12Ala of the PPAR gamma 2 gene had a significantly higher obesity risk than noncarriers (odds ratio (OR)=2.18, 95% CI=1.09-4.36) when we adjusted for sex, age and physical activity. Moreover, the risk of obesity was higher (OR=2.59, 95% CI=1.17-5.34) when family history of obesity was also taken into account in the model. The OR for obesity linked to both polymorphisms (PPAR gamma 2 and ADR beta 3) was 5.30 (95% CI=1.08-25.97) when we adjusted for sex, age and physical activity. After adjustment for family history of obesity, the OR for carriers of both polymorphisms was 19.5 (95% CI=2.43-146.8). CONCLUSIONS: A synergistic effect between polymorphism Pro12Ala of the PPAR gamma 2 gene and Trp64Arg of the ADR beta 3 gene for obesity risk was found in a case-control study including children and adolescents. PMID- 15543218 TI - Adjustable gastric banding surgery in morbidly obese adolescents: experiences with eight patients. AB - BACKGROUND: Excessive obesity in children and adolescents is increasing in western countries all over the world. Treatment of those young subjects is extremely difficult. In cases who do not respond in any regard to conservative treatment regimens, it is worthwhile to also offer bariatric surgical procedures. METHODS: In eight young subjects with a mean age of 16.0+/-1.3 y, mean BMI 49.1+/ 5.2 who where totally resistant against different therapeutic regimens, we performed adjustable laparoscopic banding surgery. Psychological tests were carried out to find out if there were any psychological disturbances. In all patients, the depression inventory for adolescents (DIKJ) and clinical interviews were carried out. RESULTS: No complications were seen; mean follow-up of 10.5+/ 6.0 months showed a weight loss of 25+/-6.5 kg. All adolescents felt for the first time that it was possible to lose weight and were very happy about their body attitude. They were able to adhere to a strict dietary regimen, which allowed them to eat without any hunger problems. It was observed that most of these patients have depressive symptoms and a very low self-esteem. Some patients suffer from nervous anxiety, poor peer acceptance, less athletic competence or forms of victimisation too. CONCLUSIONS: From these results, we conclude that laparoscopic gastric banding surgery seems to be a method that could be offered to morbidly obese adolescents. However, long-term follow-up is essential and psychological problems should be carefully studied. PMID- 15543219 TI - Programming of obesity and cardiovascular disease. AB - BACKGROUND: There is evidence that malnutrition in early life induces a growth retardation leading, in adult life, to manifest components of the metabolic syndrome. However, the impact on obesity seems less clearly established. OBJECTIVE: To review the effects of foetal and postnatal malnutrition on the programming of obesity in the context of the metabolic syndrome, as well as the link between central obesity and cardiovascular diseases. METHODS: Included in the review were recent papers exploring the mechanisms linking maternal nutrition with impaired foetal growth and later obesity, cardiovascular disease, hypertension and diabetes in humans and animals. RESULTS: The programming of obesity during foetal and early postnatal life depends of the timing of maternal malnutrition as well as the postnatal environment. Obesity arises principally in offspring submitted to malnutrition during early stages of gestation and which presented early catch-up growth. The programming may involve the dysregulation of appetite control or the hormonal environment leading to a context favourable to obesity development (hypersecretion of corticosteroids, hyperinsulinaemia and hyperleptinaemia and anomalies in the IGF axis). Adipose tissue secretes actively several factors implicated in inflammation, blood pressure, coagulation and fibrinolysis. The programmed development of intra-abdominal obesity after early growth restriction may thus favour higher prevalence of hypertension and cardiovascular diseases. CONCLUSIONS: Abdominal obesity appears in malnourished offspring and is aggravated by early catch-up growth. Higher rates of intra abdominal obesity observed after growth restriction may participate to hypertension and create atherothrombotic conditions leading to the development of cardiovascular diseases. PMID- 15543220 TI - Body composition in adolescents: measurements and metabolic aspects. AB - INTRODUCTION: Adolescence is a decisive period in human life in which important body composition changes occur. Increase of total body mass and its relative distribution are mainly related to gender and pubertal development. OBJECTIVE: This review explores the specific measurements that may be used in this age group to assess excess body fat and to define obesity and overweight. RESULTS: Identification of subjects at risk for adiposity requires simple anthropometric cutoffs for the screening of overweight and obesity. In this context, BMI criterion is the most frequently used but, in spite of its high sensitivity and specificity, an important number of adolescents classified as overweight or obese do not have really high adiposity (32.1% of females and 42% of males). Excess total body fat and intra-abdominal visceral fat are related to metabolic abnormalities that increase the risk of cardiovascular diseases. Waist circumference seems to be the best simple anthropometric predictor for the screening of the metabolic syndrome in children and adolescents. CONCLUSIONS: Early identification of adolescents at risk for adiposity and its related metabolic complications requires reliable, simple and specific measures of excess body fat for this age group. PMID- 15543221 TI - Inflammatory mediators in overweight and obese Spanish adolescents. The AVENA Study. AB - OBJECTIVE: The aim of this study was to clarify if there is an association between overweight and a state of chronic, low-grade inflammation in adolescents. DESIGN: The study is a part of the cross-sectional multicenter study AVENA, designed to evaluate the nutritional status of a representative sample of Spanish adolescents. The adolescents were divided into two groups: (1) nonoverweight and (2) overweight/obesity using the International Obesity Task Force (IOTF) standards. SUBJECTS: A geographically representative subsample of the AVENA study including 493 Spanish adolescents, aged 13-18 y (236 females/257 males), participated in this study. MEASUREMENTS: Serum C-reactive protein (CRP) and in vitro production of interleukin 6 and tumor necrosis factor alpha were measured, together with a detailed anthropometry. RESULTS: The inflammatory markers showed generally higher values in subjects with overweight/obesity than in those with nonoverweight, with only CRP showing significant differences (the means were 0.83 and 1.27 mg/l in the nonoverweight and overweight/obesity groups, respectively). CONCLUSION: Although we have not studied if adolescent overweight and obesity play an initiating role in the development of future diseases, we suggest it may induce a chronic low-grade inflammatory state, which points out the importance of maintaining an appropriate body weight, to avoid obesity-related diseases in adulthood. PMID- 15543222 TI - Metabolic risk in early childhood: the EarlyBird Study. AB - OBJECTIVE: For a decade or more, poor nutrition during gestation, expressed as low weight at birth, was held to be the factor responsible for insulin resistance later in life. Birth weights, however, are rising and insulin-resistant states, such as diabetes, faster still. Alternative explanations are needed for insulin resistance in contemporary society. This review cites data from the EarlyBird study on the relationships of insulin resistance and metabolic disturbance in early childhood. DESIGN: EarlyBird is a nonintervention prospective cohort study that asks the question 'Which children develop insulin resistance, and why?' It is unique in taking serial blood samples from a young age with which to monitor the behaviour of insulin resistance and its metabolic correlates, and in its comprehensive assessment of factors known or thought to influence insulin resistance SUBJECTS: In all, 307 randomly selected healthy school children at school entry (mean age 4.9 y) and at 12 and 24 months later. MEASUREMENTS: In the children: Birth weight and, at each time point height, weight, body mass index (BMI, kg/m(2)), skinfolds at five sites, circumferences, resting energy expenditure, physical activity, body composition, heart rate variability, diet, HOMA-IR and HOMA-ISC, blood pressure, full blood count, haemoglobin and haematocrit, HbA1C, total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, uric acid, IGF-1, gonadotrophins and SHBG. In their parents: At baseline height, weight, BMI, waist circumference, HOMA-IR and HOMA-ISC, full blood count, haematocrit, HbA1C, total cholesterol, HDL cholesterol, calculated LDL cholesterol, triglycerides, uric acid, gonadotrophins and SHBG. RESULTS: Four observations are reported here: (1) There are clear correlations in contemporary children between insulin resistance and weight at 5 y, but none with birth weight. (2) Females throughout life are intrinsically more insulin resistant than males. (3) The substantial variation of physical activity among young children is attributable to the child, and not to his environment. (4) There is dissociation in young children between fatness and insulin resistance. CONCLUSION: There is much yet to be learned about the development of obesity and insulin resistance in children. The notions of overnutrition and underactivity alone are too simplistic. PMID- 15543223 TI - The prevalence of the metabolic syndrome and type 2 diabetes mellitus in children and adolescents. AB - OBJECTIVE: To review the prevalence of the metabolic syndrome and type 2 diabetes in children and adolescents. METHOD: Literature review. RESULTS: It is well demonstrated that cardiovascular risk factors are frequent in childhood obesity and they tend to cluster. However, the frequency of the metabolic syndrome in childhood and adolescence has been investigated only by few studies. In spite of the diverse criteria used for defining the metabolic syndrome, it is evident that the syndrome is already highly prevalent among obese children and adolescents. Population-based data suggest that the epidemic of pediatric obesity is being followed by an increase of type 2 diabetes mellitus, especially in the United States and in minorities. For the European countries, there are no population based incidence and prevalence data concerning type 2 diabetes mellitus in children and adolescents. From the available data, the magnitude of the problem in the European Caucasian population seems to be much less than in North America. CONCLUSION: There is an urgent need to establish internationally acceptable criteria for the metabolic syndrome in children and adolescents and to commence screening for this syndrome. Although type 2 diabetes mellitus is still rare among European children, screening is recommended for type 2 diabetes mellitus or impaired glucose tolerance in children and especially in adolescents with substantial risk for the development of this disease. PMID- 15543224 TI - Hormones regulating lipid metabolism and plasma lipids in childhood obesity. AB - OBJECTIVE: To review the mechanisms by which leptin, insulin and adiponectin influence lipid metabolism and plasma lipids in obesity, as well as to describe the associations between these hormones in prepubertal children. METHOD: Revision of relevant papers published in the last 5 y related to the interactions of leptin, insulin and adiponectin, with special emphasis on those reporting potential mechanisms by which these hormones regulate lipid metabolism and plasma lipids. We also provide original results concerning the relationships found between plasma lipids and leptin, and insulin and adiponectin in prepubertal obese children. RESULTS: Recent data in the literature shed new light to explain the effects of both leptin and adiponectin in the regulation of lipid metabolism in peripheral tissues. Activation of the AMP-dependent kinase pathway and subsequent increased fatty acid oxidation seems to be the main mechanism of action of these hormones in the regulation of lipid metabolism. In addition, we have found that insulin plasma levels are positively associated to leptin but negatively correlated with adiponectin in obese children. Adiponectin is negatively associated to plasma lipid markers of metabolic syndrome but positively related to HDL-cholesterol, whereas insulin and leptin show opposite patterns. These results support the effect of adiponectin in increasing insulin sensitivity and decreasing plasma triglycerides. CONCLUSION: Leptin, insulin and adiponectin are associated hormones that regulate lipid metabolism in childhood. Adiponectin appears to be the missing link to explain the alterations in lipid metabolism and plasma lipids seen in obesity. PMID- 15543225 TI - The use of anthropometry in the prevention of childhood overweight and obesity. AB - OBJECTIVE: To review concepts and propose measures related to the use of anthropometry for early identification of excessive weight gain in children. METHODS: Review of results from national and international studies focusing on the assessment of childhood growth, and evaluation of the weight-for-height z scores of individual children using the 1977 National Center for Health Statistics and the 2000 Centers for Disease Control and Prevention growth charts. RESULTS: At present, few countries (23%) use indicators based on weight and height measurements to classify child body weight status. Less than one-third of growth monitoring programmes assess the growth of children beyond 6 y of age. Growth charts based on descriptive samples of populations undergoing increasing trends of childhood overweight and obesity result in substantial underestimation of true rates of these conditions. CONCLUSION: Early recognition of excessive weight gain relative to linear growth should become standard clinical practice by the following: (a) the routine collection of height measurements to enable monitoring weight-for-height and body mass index (BMI); (b) the expansion of existing monitoring programmes to include the assessment of all children up to 18 y at least once a year; (c) the interpretation of weight-for-height and BMI indices based on prescriptive reference data; and (d) the early intervention after an increase in weight-for-height or BMI percentiles has been observed. PMID- 15543226 TI - Outcome measurements in paediatric obesity prevention trials. AB - OBJECTIVE: Obesity in children impacts on their health in both the short and long term. Having an accurate and precise body composition assessment, it may be possible to control growth process and predict adult status in order to reduce the risk factors for various diseases. METHOD: To review methods for body composition assessment that may provide new insights into the clinical practicality of paediatric obesity prevention/treatment. To present which specific outcome measurements in paediatric obesity prevention trials could be used to detect subjects at risk as early as possible. RESULTS: We discussed body composition measurements that could be used in daily clinical practice and as outcome measurements in prevention trials. CONCLUSION: These measurement procedures could be associated with methods for preventing obesity onset or retarding the weight gain associated with ageing. PMID- 15543227 TI - Obesity prevention in children: opportunities and challenges. AB - OBJECTIVE: Longitudinal survey data from a number of countries confirm that the number of overweight children continues to increase at alarming rates, and even developing countries are experiencing a rise in their overweight population. There is ample consensus that prevention strategies are essential to turn the tide of the obesity epidemic, and yet there are still relatively few proven prevention approaches for children. This paper briefly discusses some of the common features of childhood obesity prevention programs, focusing on the experience in the US and Canada. APPROACH: Most prevention programs include at least one of the following components: dietary changes, physical activity, behavior and social modifications, and family participation. School-based prevention programs may also include elements related to the school environment and personnel. Primary prevention programs cannot usually restrict caloric intake, but may effectively reduce the energy intake by reducing the energy density of foods, increasing offering of fresh fruits and vegetables, using low calorie versions of products, and reducing offering of energy-dense food items. Physical activity interventions have recently focused more on reducing inactive time, particularly television viewing. Results from recent studies have reported success in reducing excess weight gain in preadolescents by restricting TV viewing. SUMMARY: Integrating all the activities of a multi-component prevention intervention, and delivering and sustaining it in different environments, continues to be a major challenge for health professionals as well as for parents, educators, and children themselves. Still, encouraging progress has been made in several areas, and the increased awareness of the problem of childhood obesity by all concerned will continue to foster our efforts in this area. PMID- 15543228 TI - Intervention centred on adolescents' physical activity and sedentary behaviour (ICAPS): concept and 6-month results. AB - OBJECTIVE: To evaluate the 6-month impact of a physical activity (PA) multilevel intervention on activity patterns and psychological predictors of PA among adolescents. The intervention was directed at changing knowledge and attitudes and at providing social support and environmental conditions that encourage PA of adolescents inside and outside school. SUBJECTS AND DESIGN: Randomised, controlled ongoing field trial (ICAPS) in middle-school's first-level adolescents from eight schools selected in the department of the Bas-Rhin (Eastern France) with a cohort of 954 adolescents (92% of the eligible students) initially aged 11.7+/-0.6 y. The 6-month changes in participation in leisure organised PA (LOPA), high sedentary (SED) behaviour (>3 h/day), self-efficacy (SELF) and intention (INTENT) towards PA were analysed after controlling for baseline measures and different covariables (age, overweight, socioprofessional occupation), taking into account the cluster randomisation design. RESULTS: The proportion of intervention adolescents not engaged in organised PA was reduced by 50% whereas it was unchanged among control students. After adjustment for baseline covariables, LOPA participation significantly increased among the intervention adolescents (odds ratio (95% confidence interval) (OR)=3.38 (1.42 8.05) in girls; 1.73 (1.12-2.66) in boys), while high SED was reduced (OR=0.54 (0.38-0.77) in girls; 0.52 (0.35-0.76) in boys). The intervention improved SELF in girls, whatever their baseline LOPA (P<10(-4)) and INTENT in girls with no baseline LOPA (P=0.04). SELF tended to improve in boys with no baseline LOPA, without reaching statistical significance. When included in the regression, follow-up LOPA was associated with improvement of SELF in girls (P=0.02) and of INTENT in girls with no baseline PA (P<0.02). The intervention effect was then attenuated. CONCLUSION: After 6 months of intervention, ICAPS was associated with a significant improvement of activity patterns and psychological predictors, indicating a promising approach for modifying the long-term PA level of adolescents. PMID- 15543229 TI - RhoB mRNA is stabilized by HuR after UV light. AB - RhoB is a small GTP-binding protein that is involved in apoptotic signal transduction. We have cloned the mouse RhoB mRNA including a 1377 nucleotide 3' untranslated region (UTR) that contains six AU-rich elements (AREs) as well as several uridine-rich stretches. There is 94% homology overall between the mouse and rat RhoB genes and 92% homology between the mouse and a putative human clone. Ultraviolet light (UVL) induces RhoB production through regulated changes in gene transcription and mRNA stabilization although the latter mechanism is unknown. We observed that UVL increased the half-life of RhoB mRNA from 63 min to 3.3 h in NIH/3T3 cells and from 87 min to 2.7 h in normal human keratinocyte cells. In vitro mobility shift assays demonstrated that HuR bound the 3'-UTR of RhoB at three distinct locations (nucleotides 1342-1696, 1765-1920 and 1897-1977) suggesting a regulatory role for this RNA-binding protein. HuR immunoprecipitations were positive for RhoB mRNA indicating an in vivo association, and Western blot analysis and immunofluorescence demonstrated that HuR rapidly partitions from the nucleus to the cytoplasm after UVL. Therefore, we propose a model in which UVL induces stress-activated signal transduction leading to nuclear/cytoplasmic shuttling of HuR and subsequent stabilization of RhoB mRNA. PMID- 15543230 TI - p12(CDK2-AP1) mediates DNA damage responses induced by cisplatin. AB - We examined the biological role of p12(CDK2-AP1) in cisplatin-mediated responses by using murine ES p12(CDK2-AP1) knockout clones generated by a targeted disruption of murine p12(CDK2-AP1). Homozygous knockout clones showed an increased cellular proliferation along with an increase in S and a decrease in G2/M phase populations. Interestingly, ES p12(CDK2-AP1) knockout clones showed a resistance to cisplatin treatment along with an increased DNA repair activity assessed by host cell reactivation assay using a cisplatin-damaged reporter DNA and a significant reduction of apoptosis upon cisplatin treatment. By using stable p12(CDK2-AP1) short interfering RNA (siRNA) clones from human normal oral keratinocytes, we confirmed that downregulation of p12(CDK2-AP1) resulted in a resistance to cisplatin. More interestingly, cisplatin treatment resulted in a reduction of CDK2 kinase activity in control clones, but p12(CDK2-AP1) knockout clones showed a sustained CDK2 kinase activity. These data suggest that p12(CDK2 AP1) plays a role in cisplatin-mediated cellular responses by modulating CDK2 activity. These data further suggest p12(CDK2-AP1) is a potential gene therapeutic agent for oral/head and neck cancer in conjunction with DNA-damaging agents such as cisplatin. PMID- 15543231 TI - The expression of TA and DeltaNp63 are regulated by different mechanisms in liver cells. AB - The TP63 gene, a member of the TP53 gene family, encodes several isoforms with (TAp63) or without (DeltaNp63) transactivating properties. Whereas the role of p63 in the normal development of squamous epithelia is well established, its function in other cell types remains to be elucidated. Here, we have analysed the expression of TA and DeltaNp63 isoforms in liver cells, by using both primary hepatocytes from wild type and p53-null mice and three human hepatocellular carcinoma (HCC) cell lines, according to the transformation state and the TP53 status of the cells. We observed the expression of DeltaNp63 isoforms only in a p53-null context. On the other hand, the expression of TAp63 isoforms was restricted to the HCC cell lines, whatever the TP53 status. We then studied the expression of TP63 upon genotoxic treatment. When treated with UVB or H(2)O(2), hepatocytes did not exhibit any change in p63 mRNA level. At the opposite, upon treatment with topoisomerase II inhibitors (doxorubicin or etoposide), the expression of TAp63 isoforms was clearly induced, independently of the TP53 status of cells. The same treatment did not induce any variation in the expression of DeltaNp63 isoforms, both at mRNA and protein levels. In HCC cell lines, doxorubicin or etoposide treatment also resulted in an increase of TAp63 transcripts only. This increase was accompanied by an increase in the intracellular level of TAp63 alpha protein. In parallel, we observed an upregulation of some p53-target genes related to cell cycle regulation, such as WAF1/CIP1, PIG3, 14-3-3sigma or GADD45, independently of the TP53 status of cells. In conclusion, we report for the first time that TA and DeltaNp63 alpha proteins are present in liver cells. Furthermore, our results suggest that p63 may partially substitute for wild-type p53, in counteracting uncontrolled liver cell proliferation in response to certain forms of DNA-damage. PMID- 15543232 TI - Efficacy and mechanism of action of the proteasome inhibitor PS-341 in T-cell lymphomas and HTLV-I associated adult T-cell leukemia/lymphoma. AB - HTLV-I associated adult T-cell leukemia (ATL) and HTLV-I-negative peripheral T cell lymphomas are associated with poor prognosis. Using pharmacological concentrations of the proteasome inhibitor PS-341, we demonstrate inhibition of cell proliferation and induction of apoptosis in fresh ATL cells, HTLV-I transformed and HTLV-I-negative malignant T cells, while normal resting or activated T lymphocytes were resistant. Combination of PS-341 and doxorubicin or etoposide resulted in an additive growth inhibition. In HTLV-I-negative malignant cells, PS-341 treatment significantly downregulated the antiapoptotic protein X IAP and to a lesser extent c-IAP-1 and bcl-X(L) and resulted in caspase-dependent apoptosis. In HTLV-I transformed cells, the inhibition of the proteasomal degradation of Tax by PS-341 likely explains the relative protection of HTLV-I infected cells against caspase-dependent apoptosis. PS-341 treatment of these cells stabilized IkappaBalpha, IkappaBbeta, IkappaBvarepsilon, p21, p27 and p53 proteins and selectively inhibited Rel-A DNA binding NF-kappaB complexes. In both HTLV-I-positive and -negative cells, PS-341 treatment induced ceramide accumulation that correlated with apoptosis. We conclude that PS-341 affects multiple pathways critical for the survival of HTLV-I-positive and -negative malignant T cells supporting a potential therapeutic role for PS-341 in both ATL and HTLV-I-negative T-cell lymphomas, whether alone or in combination with chemotherapy. PMID- 15543233 TI - Manganese superoxide dismutase overexpression inhibits the growth of androgen independent prostate cancer cells. AB - This study investigates the role of the antioxidant enzyme manganese superoxide dismutase (MnSOD) in androgen-independent human prostate cancer (PC-3) cells' growth rate in vitro and in vivo. MnSOD levels were found to be lower in parental PC-3 cells compared to nonmalignant, immortalized human prostate epithelial cells (P69SV40T). To unravel the role of MnSOD in the prostate cancer phenotype, PC-3 cells were stably transfected with MnSOD cDNA plasmid. The MnSOD protein and activity levels in clones overexpressing MnSOD were increased seven- to eightfold. These cell lines showed elongated cell doubling time, reduced anchorage-independent growth in soft agar compared to parental PC-3 (Wt) cells, and reduced growth rate of PC-3 tumor xenografts in athymic nude mice. Flow cytometric studies showed an increase in membrane potential in the MnSOD overexpressing clone (Mn32) compared to Wt and Neo cells. Also, production of extracellular H(2)O(2) was increased in the MnSOD-overexpressing clones. As determined by DNA cell cycle analysis, the proportion of cells in G(1) phase was enhanced by MnSOD overexpression. Therefore, MnSOD not only regulates cell survival but also affects PC-3 cell proliferation by retarding G(1) to S transition. Our results are consistent with MnSOD being a tumor suppressor gene in human prostate cancer. PMID- 15543234 TI - Regulation of apoptosis by fau revealed by functional expression cloning and antisense expression. AB - Functional expression cloning is a powerful strategy for identifying critical steps in biological pathways independently of prior assumptions. It is particularly suitable for the identification of molecules crucial to the control of apoptosis. Our screen for sequences suppressing T-cell apoptosis isolated a sequence antisense to fau (Finkel-Biskis-Reilly murine sarcoma virus (FBR-MuSV) associated ubiquitously expressed gene). The fox gene in FBR murine osteosarcoma virus is also antisense to fau and several reports have indicated that fau displays tumour suppressor and oncogenic properties in different contexts. Our observations indicate that the fau antisense sequence suppresses expression of endogenous fau mRNA and produces resistance to apoptosis induced both by the glucocorticoid analogue dexamethasone' by ultraviolet radiation, and by the anticancer drug cisplatin. In all cases, colony-forming ability is protected, indicating that fau affects the critical events prior to commitment to cell death. Overexpression of fau in the sense orientation induces cell death, which is inhibited both by Bcl-2 and by inhibition of caspases, in line with its proposed role in apoptosis. PMID- 15543235 TI - Demonstration of tumor suppression by mannose 6-phosphate/insulin-like growth factor 2 receptor. AB - The mannose 6-phosphate/IGF-2 receptor has been proposed to be a tumor suppressor gene on the basis of loss of heterozygosity and mutations in tumors from cancer patients. To test this hypothesis, the receptor was expressed in 66cl4, a mouse mammary tumor cell line deficient in the receptor. Expression of the receptor corrected the abnormal lysosomal trafficking phenotype displayed by these cells. Receptor expression had no apparent effect on growth or invasiveness of the cells in vitro but effectively inhibited formation of mammary tumors in BALB/c mice. Analysis of cell proliferation and apoptosis in tumors indicated that the primary effect of the receptor was to inhibit cell proliferation. Proliferation indices for receptor-deficient and receptor-expressing tumors, as determined by BrdU incorporation, were 24.6 and 7.6%, respectively. No significant effect of receptor expression on apoptosis was observed. Receptor expression similarly inhibited tumor growth in BALB/c scid mice indicating that cytotoxic T cells and other components of the immune system missing in scid mice are not involved in the receptor's tumor suppressing effect. These findings establish a role for the receptor as a bona fide tumor suppressor gene and together with previous studies, suggest an important role for the receptor in human and rodent cancers. PMID- 15543236 TI - Ras-dependent induction of HIF-1alpha785 via the Raf/MEK/ERK pathway: a novel mechanism of Ras-mediated tumor promotion. AB - Hypoxia-inducible factor-1alpha (HIF-1alpha) plays crucial roles in tumor promotion by transactivating approximately 60 kinds of its target genes. Recently, we reported a novel splice variant HIF-1alpha(785), which is regulated primarily by phorbol ester. This variant can be stabilized under normoxic conditions because it loses an acetylation site Lys532. Its expression was found to promote xenografted tumor growth in nude mice. We here found that the Ras oncogene regulates HIF-1alpha(785) expression via the Raf/MEK/ERK pathway, and that both phorbol ester and epidermal growth factor also induced HIF-1alpha(785) via the same pathway. We also identified the nonhypoxic regulatory domain responsible for phorbol ester-induced HIF-1alpha(785) expression. These results imply that HIF-1alpha(785) may play an important role in tumor promotion mediated by the Ras oncogene, phorbol ester or tumor growth factors. PMID- 15543237 TI - Identification of new classes among acute myelogenous leukaemias with normal karyotype using gene expression profiling. AB - Conventional cytogenetic analysis currently stratifies acute myelogenous leukaemia (AML) into prognostically relevant groups. However, approximately 50% of adult AMLs have normal cytogenetics (NC-AMLs), and represent a heterogeneous and poorly understood group. We analysed gene expression in 55 AML samples including 53 cases from adult patients with NC-AML (n = 36), trisomy 8, t(15;17), t(8;21), t(11;19), 7q deletion, and two cell lines using 9000-gene DNA microarrays. Global hierarchical clustering showed that NC-AMLs are a heterogeneous group. Supervised analysis distinguished two subgroups of NC-AML: one subgroup constituted a homogeneous NC cluster ('pure NC-AML'), and the other NC-AMLs were close to the AML cases with translocations ('translocation like'). Gene expression signatures were also derived for patients with trisomy 8, as well as FLT3 and MLL gene duplications. Importantly, samples from 24 NC-AML patients who could be evaluated for clinical outcome were analysed. In all, 43 genes that discriminated two classes of patients with significantly different prognosis were identified. The poor prognosis class contained a majority of 'pure NC-AMLs', whereas the 'translocation-like' AMLs were in the good prognosis class. Discriminator genes included genes involved in drug resistance (TOP2B), protein transport (MTX2, SLC35A2), and cell signalling (MAPK1, PRKAB2). Our results demonstrate the transcriptional heterogeneity of NC-AMLs, and suggest the existence of 'translocation-like' NC-AMLs and of a gene expression signature that may predict response to chemotherapy. PMID- 15543238 TI - Tobacco control in Australia: victims of our past success? PMID- 15543239 TI - Current tobacco smoking by the New South Wales population and the consequences for health. PMID- 15543240 TI - The social costs of smoking in Australia. PMID- 15543241 TI - A report on the New South Wales Tobacco Action Plan 2001-2004. PMID- 15543243 TI - Litigation and its current role in tobacco regulation in Australia. PMID- 15543242 TI - Managing nicotine dependence in New South Wales hospital patients. PMID- 15543244 TI - Promoting tobacco to the young in the age of advertising bans. PMID- 15543245 TI - The 2004 United States Surgeon General's Report: The Health Consequences of Smoking. PMID- 15543246 TI - The impact of low tar cigarettes. PMID- 15543247 TI - Tobacco and health fact sheet: light cigarettes. PMID- 15543248 TI - Continuous New South Wales Health Survey: quarterly report on health status, health behaviours, and risk factors. PMID- 15543249 TI - Communicable diseases report, New South Wales, for March and April 2004. PMID- 15543251 TI - Alternative treatments for disseminated intravascular coagulation. AB - The acronym DIC is commonly interpreted as "death is coming." This pessimistic view emphasizes the deficiency of available treatment options following diagnosis of disseminated intravascular coagulation. Clinically, DIC manifests as a systemic hemorrhagic disorder associated with widespread activation and eventual exhaustion of the coagulation system, although events underlying DIC also involve effectors of inflammation. DIC can be associated with diverse conditions including sepsis and major trauma and, when identified, signifies a significant worsening in prognosis and expected mortality. Although recent clinical studies have shown that activated protein C reduces mortality in patients with severe sepsis, there is a need for further investigation and a better understanding of the underlying mechanisms. PMID- 15543252 TI - Advances in the treatment of hypertension: a report from the XIV Annual Meeting of the European Society of Hypertension. PMID- 15543254 TI - Treatment of hypertension in 2004: new research disclosed during the Annual Meeting of the American Society of Hypertension. PMID- 15543255 TI - Severe heart failure. PMID- 15543256 TI - Moving drug therapies in cardiology: summary of presentations at the American College of Cardiology Scientific Session 2004. PMID- 15543257 TI - Highlights from the 74th Congress of the European Atherosclerosis Society. PMID- 15543259 TI - Moving cardiovascular pharmacotherapy: a report of poster presentations during the European Society of Cardiology 2004 Annual Congress. PMID- 15543260 TI - Cholesterol-lowering with statins: are they all the same? - Summary of the Expert Symposium at the 53rd Annual Scientific Sessions of the American College of Cardiology. AB - Since they share a common mode of action and certain structural features, all statins exhibit the same broad pharmacological effects. Nevertheless, important differences exist between the statins in terms of LDL cholesterol-lowering, endpoint reduction, pleiotropic effects and toxicity. For this reason, it is incorrect to view all statins as being interchangeable as lipid-lowering therapy. In addition, it should be remembered that perhaps the main difficulty in clinical practice today does not concern details of the differences between statins but rather the fact that in many studies long-term compliance has been shown to be only around 30%. PMID- 15543261 TI - Electrostatic interactions by design. Versatile methodology towards multifunctional assemblies/nanostructured electrodes. AB - This Feature Article describes how multi-site interactions between positively or negatively charged carbon forms, such as fullerenes and single wall carbon nanotubes, and porphyrinic chromophores have been utilized en route towards novel multifunctional and nanostructured materials. Specifically, we discuss (i) the behavior of molecular assemblies in homogeneous solutions and (ii) the controlled self-assembly on surfaces. PMID- 15543262 TI - Proton transfer and N(+)-H...S(-) hydrogen bonds in the crystal structure of 4 aminothiophenol. AB - 4-Aminothiophenol exists as 4-ammonio-1-benzenethiolate in the solid and liquid state. The crystal structure is characterised by a tetrahedral beta-As type network which is the driving force for the proton transfer. PMID- 15543263 TI - Crystal engineering in the aminophenols. Novel carborundum network in a supramolecular homologous series. AB - Molecular complexes of 4-(4-aminophenoxy)aniline with a series of diphenols are structurally homologous and adopt the carborundum III topology, which is an unprecedented network for organic solids. PMID- 15543264 TI - Capturing a metastable chiral polymorph of an achiral molecule--hexa-O-benzoyl myo-inositol. AB - myo-Inositol hexabenzoate having meso configuration produces chiral polymorph (form I) when crystallized rapidly but yields achiral polymorph (form II) when allowed to crystallize slowly; in the mother liquor form I slowly but completely disappears to give form II. PMID- 15543265 TI - Photon-driven nanomechanical cyclic motion. AB - Microcantilevers modified by a monolayer of azobenzene molecules bend up and down periodically, switched by a 365 nm UV light, as a result of the conversion of the two configurations of azobenzene molecules in the monolayer. PMID- 15543266 TI - Sextuplet phenyl embrace in a metal-organic Kagome lattice. AB - A novel Kagome lattice that demonstrates the modular nature of metal-organic networks has been prepared and is to our knowledge the first example of a metal organic coordination polymer that incorporates the sextuplet phenyl embrace as a supramolecular synthon. PMID- 15543267 TI - Highly efficient bromination of aromatic compounds using 3-methylimidazolium tribromide as reagent/solvent. AB - 3-Methylimidazolium tribromide proves to be an alternative highly efficient reagent/solvent for the halogenation of non-activated aromatic compounds. PMID- 15543268 TI - Photoactive dimesogen having different pathways of light driven phase transitions at different temperatures. AB - We hereby describe a novel example of light driven phase transitions in a photoresponsive dimesogen, which goes through different pathways at different temperatures. The isomer ratios required to obtain the phase transition in each case have also been determined. PMID- 15543269 TI - Mo2P4O15--the most complex oxide structure solved by single crystal methods? AB - We report the crystal structure and phase transitions of Mo2P4O15 which, despite a simple chemical formula, has 441 crystallographically unique atoms in its asymmetric unit and thus has the most complex structure of any extended oxide reported to date. PMID- 15543270 TI - Chiral self-dimerization of vanadium complexes on a SiO2 surface: the first heterogeneous catalyst for asymmetric 2-naphthol coupling. AB - The self-dimerized chiral assembly of vanadium-Schiff-base complexes was found to occur on a SiO2 surface and to be the first heterogeneous catalyst for the asymmetric oxidative coupling of 2-naphthol with 100% selectivity and 90% enantioselectivity. PMID- 15543271 TI - Development of porous materials for heterogeneous catalysis: kinetic resolution of epoxides. AB - Template copolymerization methods have been utilized to prepare porous materials with immobilized cobalt complexes that catalyze the hydrolytic kinetic resolution of epoxides. PMID- 15543272 TI - Carene terpenoids by gold-catalyzed cycloisomerization reactions. AB - Propargyl acetates in the presence of catalytic amounts of AuCl3 constitute synthetic equivalents of alpha-diazoketones as illustrated by a concise entry into the carene family of natural products. PMID- 15543273 TI - Co(II)5(OH)6(SO4)2(H2O)4: the first ferromagnet based on a layered cobalt hydroxide pillared by inorganic...OSO3-Co(H2O)4-O3SO... AB - The synthetic mineral Co(II)5(OH)6(SO4)2(H2O)4 (1), obtained by hydrothermal reaction of CoSO4.7H2O and NaOH at 165 degrees C and consisting of brucite-like Co4(OH)6O2 layers pillared by OSO3-Co(H2O)4-O3SO, is a ferromagnet (T(Curie)= 12 K, Hc= 580 Oe). PMID- 15543274 TI - Enhanced selectivity in non-heme iron catalysed oxidation of alkanes with peracids: evidence for involvement of Fe(IV)=O species. AB - Catalytic alkane oxidation with high selectivity using peracids and an (N4Py)Fe complex is presented and the role of [(N4Py)Fe(IV)=O]2+ species, molecular oxygen and hydroxyl radicals in the catalysis is discussed. PMID- 15543275 TI - Biomimetic synthesis of the pyrrolobenzoxazine core of paeciloxazine. AB - Starting from a protected L-tryptophan derivative the pyrrolobenzoxazine core unit of paeciloxazine can be synthesized in two oxidation steps. PMID- 15543276 TI - Hierarchically ordered CdS doped nanoporous membrane. AB - Large-dimension heterogeneous nanoporous membranes were obtained by the formation of CdS doped mesoporous silica within the porous channels of alumina membranes. PMID- 15543277 TI - The substituent-dependent base-treatment chemistry of (eta5 C5H5)Fe(CO)2SiR2SiR2Cl: formation of 1,2-disila-3-metallacyclobutanes, their ring opened polymers, migrations and substitutions. AB - We report the formation of disilametallacycles, high molecular weight polymers or simple substitution products from the reactions of 1-(cyclopentadienyliron dicarbonyl)-2-chlorodisilanes with bases, LDA or n-BuLi. PMID- 15543278 TI - Sonochemical synthesis of SnO2 nanobelt/CdS nanoparticle core/shell heterostructures. AB - SnO2 nanobelt/CdS nanoparticle core/shell heterostructures are successfully achieved via a simple sonochemical approach; their ethanol-sensing properties are discussed. PMID- 15543279 TI - Magnetic nanoparticle assemblies on denatured DNA show unusual magnetic relaxivity and potential applications for MRI. AB - Denatured (substantially single-stranded) herring sperm DNA acts as a template for the preparation of magnetic nanowires, forming stable aqueous suspensions, which exhibit unprecedentedly high relaxivity at low field, suggesting that the material may be a potentially useful reagent for MRI. PMID- 15543280 TI - Oxide surface-promoted Pd-complex catalysis for intramolecular O-activated alkene hydroamination: catalyst preparation, characterization, and performance. AB - Supported Pd-P and Pd-N complexes prepared by a selective reaction of Pd-monomer precursors with SiO2, Al2O3 and TiO2 surfaces and characterized by EXAFS, XPS, XRF and gas analysis exhibited oxide surface-promoted catalysis for the catalytic intrahydroamination of 3-amino propanol vinyl ether to produce a cyclic amine. PMID- 15543281 TI - An N,P-disubstituted-2-aminophosphaalkene and lithium and potassium complexes of the deprotonated "phosphaamidinate" anion. AB - The reaction of DippPH2(Dipp = 2,6-iPr2C6H3) with DippN=C(p-CH3C6H4)Cl in refluxing xylenes affords DippP=C(p-CH3C6H4)N(H)Dipp; deprotonation with alkali metal reagents produces unique lithium and potassium complexes with the ligand in a different geometry to that of the free phosphaamidine. PMID- 15543282 TI - Controlling stereochemistry during oxidative coupling. Preparation of Rp or Sp phosphoramidates from one P-chiral precursor. AB - Stereochemical outcome of oxidative coupling of H-phosphonate diesters with amines, promoted by iodine, can be controlled to obtain the corresponding phosphoramidate diesters with inversion or with retention of configuration at the phosphorus centre. PMID- 15543283 TI - Bidentate amidine ligands for nickel(0)-mediated coupling of carbon dioxide with unsaturated hydrocarbons. AB - Novel bidentate amidines were designed and synthesized as easily available electron-donating N-ligands for Ni0-mediated coupling of carbon dioxide with alkynes or allenes, and high regioselectivity was achieved even for the carboxylation of aryl substituted internal alkynes. PMID- 15543284 TI - Cofactor-dependent enzyme catalysis in functionalized ionic solvents. AB - Functionalized, hydrogen-bonding ionic liquids have been successfully evaluated as media for the performance of cofactor-dependent enzyme catalysed oxidations; the effects of incorporating hydroxyl groups into both the cation and anion have been studied and the dependence of activity upon water content has been evaluated. PMID- 15543285 TI - Ethanol vapor-mediated maturing for the enhancement of structural regularity of hexagonal mesoporous silica films. AB - Structural regularity of hexagonal mesoporous silica films is dramatically enhanced when allowed to stand for a few hours in an ethanol vapor-containing closed bottle before dryness. PMID- 15543286 TI - NRIS study on the [FeN6] core in photo-induced high-spin state of [Fe(2 pic)3]Cl2.EtOH. AB - A comparison is reported for thermally and photo-induced high-spin phases of [Fe(2-pic)3]Cl2.EtOH (2-pic: 2-picolylamine) using the nuclear resonant inelastic scattering (NRIS) technique. PMID- 15543287 TI - Novel pitch-based carbons with bimodal distribution of uniform mesopores. AB - A new method is proposed for the synthesis of pitch-based carbons with bimodal distribution of uniform mesopores formed by co-imprinting of spherical silica colloids and hexagonally ordered mesoporous particles of SBA-15 into mesophase pitch particles and subsequent silica dissolution. PMID- 15543288 TI - Red luminescent polymeric cuprous organosulfide generated by solvothermal redox reaction. AB - A new polymeric metal organosulfide [Cu(I)3(pymt)3]n(pymt = pyrimidine-2 thiolate) with strong red photoluminescence was synthesized through solvothermal redox reaction, and crystallographically characterized to be a one-dimensional chiral structure containing metal-metal interactions. PMID- 15543289 TI - 'Double asymmetric induction' as a novel tool for high stereocontrol in Baylis Hillman reaction. AB - The strategy of double asymmetric induction was utilized in Baylis-Hillman reaction for the first time by the coupling of chiral aldehydes with chiral acrylate (1,2:5,6-di-O-isopropylidene-alpha-D-glucofuranose-3-acrylate) to obtain corresponding adducts with high syn diastereoselectivities (de >90%) in moderate to good yields. PMID- 15543290 TI - Synthesis of Ag2S quantum dots in water-in-CO2 microemulsions. AB - Ag2S nanocrystals with a mean diameter of 5.9 nm (sigma= 1.65 nm) and characteristic surface plasmon resonance absorption at 330 nm have been synthesized in water-in-supercritical CO2 reverse microemulsion using the commonly used AOT surfactant with 2,2,3,3,4,4,5,5-octafluoro-1-pentanol (F pentanol) as cosurfactant. PMID- 15543291 TI - A samarium cyclooctatetraene complex as catalyst for hydroamination/cyclisation catalysis. AB - The cyclooctatetraene-bis(phosphinimino)methanide complex [[CH(PPh2NSiMe3)2]Sm(eta8-C8H8)] has been prepared; although this compound has no alkyl or amide ligand it shows moderate activity as a catalyst for the hydroamination/cyclisation reaction. PMID- 15543292 TI - Electrophilicity parameters for sigma-complexation by uncharged electron deficient aromatic and heteroaromatic structures. AB - Using appropriate sets of reference nucleophiles, the reactivity of neutral electrophiles of widely different reactivity and structure has been ranked on the comprehensive electrophilicity scale of Mayr (Acc. Chem. Res., 2003, 36, 66), holding promise of a general rationalization of sigma-complexation processes and related SNAr substitutions. PMID- 15543293 TI - An unprecedented intramolecular coupling of o-carboranyl and cyclopentadienyl. Synthesis and structural characterization of a ruthenium complex containing a novel doubly-bridged cyclopentadienyl-carboranyl ligand. AB - A novel ruthenium-mediated coupling reaction of a carboranyl with a cyclopentadienyl was discovered for the first time, leading to a brand new doubly bridged cyclopentadienyl-carboranyl ligand bearing two five-membered rings and to a new type of chiral ruthenium complex. PMID- 15543294 TI - A mixed ladderane/n-alkyl glycerol diether membrane lipid in an anaerobic ammonium-oxidizing bacterium. AB - A novel glycerol diether containing ladderane and tetradecyl moieties has been identified in an anaerobic ammonium-oxidizing bacterium by GC/MS and high-field NMR spectroscopy. PMID- 15543295 TI - Bundles of identical double-walled carbon nanotubes. AB - In a sample produced by catalytic chemical vapor deposition (CCVD), the structure of the carbon nanotubes (diameter and helicity) which governs their electronic properties, is determined by electron diffraction. We found that most of the smallest bundles are constituted of identical double-walled carbon nanotubes. PMID- 15543296 TI - Novel fullerene-porphyrin-fullerene triad linked by metal axial coordination: Synthesis, X-ray crystal structure, and spectroscopic characterizations of trans bis([60]fullerenoacetato)tin(IV) porphyrin. AB - A novel fullerene-porphyrin-fullerene triad linked by the axial coordination of tin(IV) porphyrin is reported, exhibiting strong pi-pi interactions between the tin porphyrin and fullerene moieties owing to their close proximity. PMID- 15543297 TI - Aminooxazolinate; a chiral amidinate analogue. AB - High levels of diastereoselection with respect to chirality-at-metal are achieved at equilibrium for complexes containing a new and available range of diazaallyl ligands. PMID- 15543298 TI - A simple, general and efficient ketone synthesis via alkylation and dephosphinoylation of beta-keto-diphenylphosphine oxides. AB - Products of difficult ketone alkylation reactions can be made selectively via activation with a diphenylphosphinoyl group; subsequent dephosphinoylation is easily achieved in base. PMID- 15543299 TI - Biosynthetic studies on the azinomycins: the pathway to the naphthoate fragment. AB - Isotopically labelled intermediates in a proposed biosynthesis of the naphthoate fragment of azinomycin B have been made and successfully incorporated by the azinomycin producing organism. PMID- 15543300 TI - Lanthanide(III) complexes of a pyridine N-oxide analogue of DOTA: exclusive M isomer formation induced by a six-membered chelate ring. AB - The presence of a six-membered chelate ring involving a pyridine N-oxide moiety induces exclusive M isomer formation throughout the whole lanthanide series endowed with a fast water exchange in the case of the Gd(III) complex. PMID- 15543301 TI - Site-specific ligation of anthracene-1,8-dicarboxylates to an Mn12 core: a route to the controlled functionalisation of single-molecule magnets. AB - A novel single-molecule magnet of the Mn12 family, [Mn12O12(O2CC6H5)8(L)4(H2O)4].8CH2Cl2, has been synthesised by site-specific ligand exchange using a tailor-made dicarboxylate (L2-), which leads to selective occupation of axial binding sites. PMID- 15543302 TI - Stereoconservative Negishi arylation and alkynylation as an efficient approach to enantiopure 2,2'-diarylated 1,1'-binaphthyls. AB - Negishi arylation and alkynylation of easily synthesized chiral 2,2'-diodo-1,1' binaphthyl rapidly proceeds in refluxing THF utilizing controlled microwave irradiation, affording enantiopure 2,2'-diarylated 1,1'-binaphthyls in good to excellent yields. PMID- 15543303 TI - A photo-responsive organogel. AB - A photo-responsive organogel has been made by addition of a novel stilbene containing photo-surfactant to toluene: exposure to UV light led to a gel-to-sol transition with spatial control. PMID- 15543304 TI - Catalyst surface characterization in microfabricated reactors using pulse chemisorption. AB - The metal dispersion of a Pt-Al2O3 catalyst was measured reproducibly using pulse CO chemisorption with 4 mg of sample in a silicon microfabricated packed-bed reactor, demonstrating the applicability of micoreactors for high-throughput catalyst characterization with quantitative comparison. PMID- 15543305 TI - Ordered mesoporous titanosilicates with catalytically stable and active four coordinated titanium sites. AB - The stable ordered mesoporous titanosilicate (Ti-JLU-20) has been successfully synthesized from an assembly of mixed surfactants (fluorocarbon and triblock copolymer surfactants) with preformed titanosilicate zeolite precursors at high temperature (180-220 degrees C), and catalytic tests show that Ti-JLU-20 has highly stable and active four-coordinated titanium sites in oxidations. PMID- 15543306 TI - Effective and selective iodofunctionalisation of organic molecules in water using the iodine-hydrogen peroxide tandem. AB - Efficient hydrogen peroxide enhanced iodofunctionalisation of ketones, 1,3 dicarbonyl derivatives and activated aromatic molecules using elemental iodine in water is achieved, whereas alkynes were stereoselectively converted into (E)-1,2 diiodoalkenes. PMID- 15543307 TI - New reactions in water: metal-free conversion of alcohols and ketones into alpha iodoketones. AB - Using water as the reaction medium, ketones can be transformed into alpha iodoketones upon treatment with sodium iodide, hydrogen peroxide and an acid; interestingly, alpha-iodoketones can be also obtained from secondary alcohols through a metal-free tandem oxidation-iodination approach. PMID- 15543308 TI - SSZ-60: a new large-pore zeolite related to ZSM-23. AB - SSZ-60 is a new borosilicate zeolite that may be prepared using N-ethyl-N-(2,4,4 trimethylcyclopentyl)pyrrolidinium or N-ethyl-N-(3,3,5 trimethylcyclohexyl)pyrrolidinium as a structure directing agent; the framework of SSZ-60 was determined by the FOCUS Fourier recycling method, its structure refined in space group P21nm and found to possess a one-dimensional channel system with pores delimited by twelve-rings; the topological structure of SSZ-60 may be derived from a sigma-expansion of the framework of ZSM-23 (MTT). PMID- 15543309 TI - Role of pore curvature on the thermal stability of gold nanoparticles in mesoporous silica. AB - Pores arranged in a two-dimensional hexagonal structure inside spherical mesoporous silica particles help to prevent the thermal sintering of gold nanoparticles compared to straight pores in MCM-41. PMID- 15543310 TI - Nitrile oxide cycloadditions in supercritical carbon dioxide. AB - The regioselectivity of dipolar cycloadditions of mesitonitrile oxide to various dipolarophiles in supercritical carbon dioxide can be tuned by changes in density, the magnesium bromide-mediated cycloaddition to pent-1-en-3-ol proceeding with higher stereoselectivity than in most conventional solvents. PMID- 15543311 TI - Short and stereoselective synthesis of C-glycosylated glycine derivatives from glycals by radical addition and reduction. AB - Only three steps are required for the convenient synthesis of 2-C-branched glyco amino acids from glycals with good yields and stereoselectivities. PMID- 15543312 TI - [Li(thf)3cyclo-(P4tBu4CH)]--synthesis, molecular structure and dynamic behaviour. AB - [Li(thf)3cyclo-(P4tBu4CH)] (2-Li), containing the first tetraphosphacyclopentanide anion cyclo-(P4tBu4CH)- (2), was prepared, and its dynamic behaviour in solution analysed by variable-temperature 31P NMR spectroscopy. PMID- 15543313 TI - Unprecedented Barbier-type reactions catalysed by titanocene(III). AB - Selective Barbier-type allylations, benzylations and propargylations of aldehydes and ketones can be carried out under extremely mild conditions employing titanocene(III) complexes as catalysts. In this way, chiral titanocene catalysts provided yields ranging from 50-80% of optically active products. PMID- 15543314 TI - Highly efficient epoxidation method of olefins with hydrogen peroxide as terminal oxidant, bicarbonate as a co-catalyst and oxodiperoxo molybdenum(VI) complex as catalyst. AB - A combination of the newly synthesized and structurally characterized compound, [MoO(O2)2(saloxH)](saloxH2= salicylaldoxime) as catalyst, H2O2 as terminal oxidant and NaHCO3 as co-catalyst when stirred in CH3CN (10 cm3) at room temperature (rt) shows a very pronounced efficiency epoxidation of olefinic compounds, the method being green and economical. PMID- 15543315 TI - The Thr715Pro single nucleotide polymorphism of P-selectin: does it really matter in cardiovascular or thrombotic disorders? PMID- 15543316 TI - The story of the serpin plasminogen activator inhibitor 1: is there any need for another mutant? AB - The importance of obtaining insight in the structure/function relationship in the serpin plasminogen activator inhibitor type-1 can be understood from the major role PAI-1 plays in different (patho)physiological processes, mainly because of its involvement in the plasminogen/plasmin system. Moreover, during the past years, studies indicated a contribution of PAI-1 to the development of cardiovascular disease in common syndromes such as atherosclerosis, diabetes and hypertension. Furthermore, PAI-1 also inhibits u-PA, attributing a role in phenomena such as cell migration and tissue remodelling. Considering the role of PAI-1 in such various pathogenic path-ways, detailed insight into the structure/function relationship in PAI-1 might provide a means of interfering with a given pathological situation without disturbing other physiological processes. Therefore, since the discovery of PAI-1 and the cloning of its cDNA 20 years ago, over 600 PAI-1 variants have been constructed, elucidating the most important structural features of PAI-1. This review gives an overview of the contribution of the different PAI-1 variants to the understanding of the structure/function relationship in PAI-1, based on the different functional features of PAI-1. PMID- 15543317 TI - Cyclooxygenase COX-2a, a novel COX-2 mRNA variant, in platelets from patients after coronary artery bypass grafting. AB - There are two principal cyclooxygenase isoforms referred to as COX-1 and COX-2. Recently, COX-3 has been identified. We have demonstrated the expression of COX-2 in platelets from patients after coronary artery bypass grafting (CABG). Careful biochemical analysis revealed that, when compared to recombinant COX-2, platelet COX-2 had a slightly higher electrophoretic mobility. Two COX-2 sequences (approximately 1.8 kb, approximately 1.7 kb) were cloned from platelet mRNA. The approximately 1.7 kb sequence, designated COX-2a, differed from the human COX-2 sequence only in a deletion from position +458 to +567. Similar to the human COX 3, there is a frame shift in the COX-2a sequence resulting in a TAA stop codon at position +490. Thus, the expression of a COX-2a protein corresponding to the 67 kDa COX-2 protein is not clear. However, the marked shifting from COX-2 to COX-2a in platelets from some patients after CABG is a striking finding. PMID- 15543318 TI - Accelerating ability of synthetic oligosaccharides on antithrombin inhibition of proteinases of the clotting and fibrinolytic systems. Comparison with heparin and low-molecular-weight heparin. AB - The abilities of three synthetic oligosaccharides to accelerate antithrombin inhibition of ten clotting or fibrinolytic proteinases were compared with those of unfractionated, fractionated high-affinity and low-molecular-weight heparins. The results show that the anticoagulant effects of the latter three heparins under conditions approximating physiologic are exerted almost exclusively by acceleration of the inactivation of thrombin, factor Xa and factor IXa to near diffusion-controlled rate constants of approximately 10(6) - 10(7) M(-1).s(-1). All other proteinases are inhibited with at least 20-fold lower rate constants. The anti-coagulant ability of the synthetic regular (fondaparinux) and high affinity (idraparinux) pentasaccharides is due to a common mechanism, involving acceleration of only factor Xa inhibition to rate constants of approximately 10(6) M(-1).s(-1) . A synthetic hexadecasaccharide, containing both the pentasaccharide sequence and a proteinase binding site, exerts its anticoagulant effect by accelerating antithrombin inactivation of both thrombin and factor Xa to rate constants of approximately 10(6) - 10(7) M(-1).s(-1), although thrombin appears to be the more important target. In contrast, factor IXa inhibition is appreciably less stimulated. The conformational change of antithrombin induced both by the pentasaccharides and longer heparins contributes substantially, approximately 150-500-fold, to accelerating the inactivation of factors Xa, IXa and VIIa and moderately, approximately 50-fold, to that of factor XIIa and tissue plasminogen activator inhibition. The bridging effect due to binding of antithrombin and proteinase to the same, long heparin chain is dominating, approximately 1000-3000-fold, for thrombin inhibition and is appreciably smaller, although up to approximately 250-350-fold, for the inactivation of factors IXa and XIa. These results establish the proteinase targets of heparin derivatives currently used in or considered for thrombosis therapy and give new insights into the mechanism of heparin acceleration of antithrombin inhibition of proteinases. PMID- 15543319 TI - Effects of heparin and low-molecular-weight heparins on bone mechanical properties in rats. AB - Long-term administration of heparin can be associated with development of osteoporosis. The present study was carried out to compare the effects of standard heparin and two low-molecular-weight heparins (nadroparin and enoxaparin) on bone mechanical properties in rats. The experiment was conducted on 3-month-old female Wistar rats. Standard heparin was administered in doses of 1000 or 2000 IU/kg sc daily, nadroparin and enoxaparin were administered in doses of 1000 or 2000 anti-Xa IU/kg sc daily, for 28 days. Bone mass, length, diameter, mineral content and mechanical properties of the whole femur (extrinsic stiffness, ultimate and breaking load, deformation caused by the applied load) and the femoral neck (load at fracture) were examined. Standard heparin (2000 IU/kg sc daily) weakened the femoral neck. Enoxaparin and the higher doses of standard heparin and nadroparin induced similar unfavourable changes in mechanical properties of the whole femur; the changes were noticeable the most after administration of enoxaparin. PMID- 15543320 TI - Herpes simplex virus type 1-encoded glycoprotein C enhances coagulation factor VIIa activity on the virus. AB - Tissue factor (TF) is the blood coagulation initiator, whose cofactor function is required for physiological factor VIIa (FVIIa)-mediated activation of factor X (FX) to FXa. A previous study reported TF on herpes simplex virus type 1 (HSV1), but this explained only part of FVIIa-dependent FXa generation observed on the virus surface (Sutherland et al. (1997) Proc. Natl. Acad. Sci. USA. 94:13510-14). In the current study, we investigated the role of HSV1-encoded glycoprotein C (gC) in this process. Purified gC-deficient HSV1 facilitated several fold less FX activation by FVIIa than either wild type or gC-rescued strains. To confirm the implication of gC in FVIIa-dependent FX activation, purified soluble gC (sgC) enhanced FXa production in the absence of TF. sgC required FVIIa, calcium and anionic phospholipid to participate in FX activation, suggesting similarity to TF. When purified virus was combined with sgC, the sgC-dependent FXa generation was enhanced three orders of magnitude, suggesting synergy with an additional HSV1 component and explaining the relatively low activity of purified sgC compared to the viral counterpart. FX activation on gC-competent HSV1 was inhibited 20% by a gC-specific antibody, inhibited 40% by a TF-specific antibody, inhibited 65% by combining the gC- and TF-specific antibodies, and nearly completely inhibited by the TF antibody alone on gC-deficient HSV-1. Cumulatively, these observations show that two pathways initiating FX activation function in parallel on the virus surface. In addition to the previously described TF-dependent pathway, HSV-1-encoded gC also enhances FXa generation, and like TF, requires FVIIa. PMID- 15543321 TI - P-selectin-targeting of the fibrin selective thrombolytic Desmodus rotundus salivary plasminogen activator alpha1. AB - During thrombosis, P-selectin is expressed on the surface of activated endothelial cells and platelets. We hypothesized that targeting a plasminogen activator (PA) to P-selectin would enhance local thrombolysis and reduce bleeding risk. Previously, a urokinase (uPA)/anti-P-selectin antibody (HuSZ51) fusion protein was shown to increase fibrinolysis in a hamster pulmonary embolism model. To explore the therapeutic potential of this targeting strategy, we fused the fibrin-selective Desmodus rotundus salivary PA alpha1 (dsPA alpha 1) to HuSZ51 and compared the fibrinolytic activity of P-selectin-targeted dsPA alpha 1 (HuSZ51-dsPA alpha 1) to unmodified dsPA alpha 1 in vitro and in vivo. HuSZ51 dsPA alpha 1 and dsPA alpha 1 were expressed in CHO cells and purified to homogeneity by affinity chromatography. HuSZ51-dsPA alpha 1 bound to thrombin activated human and dog platelets with comparable affinities to that of parental antibody SZ51. The fusion protein retained the catalytic activities of dsPA alpha 1 in chromogenic and clot lysis assays, indicating that dsPA alpha 1 is fully functional when fused to HuSZ51. Compared to dsPA alpha 1, HuSZ51-dsPA alpha 1 had similar thrombolytic efficacy in a rat pulmonary embolism model and anti thrombotic potency in a dog model of femoral artery thrombosis. However, HuSZ51 dsPA alpha 1 was less effective in lysis of preexisting arterial thrombi in the dog model. The reduced arterial thrombolysis was not due to the pharmacokinetic properties of HuSZ51-dsPA alpha 1 because antigen level and amidolytic activity were higher in plasma from HuSZ51-dsPA alpha 1-treated groups than corresponding dsPA alpha 1-treated groups. These data indicate that the thrombolytic efficacy of HuSZ51-dsPA alpha 1 varied dependent on the physical composition of thrombi. The lack of stimulation by fibrin in arterial thrombi may contribute to the attenuated thrombolytic efficacy of HuSZ51-dsPA alpha 1 in the dog model. PMID- 15543322 TI - Temporal changes in pulmonary expression of key procoagulant molecules in rabbits with endotoxin-induced acute lung injury: elevated expression levels of protease activated receptors. AB - Recently a new concept has emerged implicating thrombin signaling as the "bridge" that connects tissue damage to hemostatic and inflammatory responses. In view of this concept, we hypothesized that induction of protease-activated receptor (PAR) expression may play a critical role in endotoxin-induced tissue injury through the cellular actions of thrombin. Thus, in this study, temporal changes in expression of key precoagulant molecules, including PARs, in lungs from rabbits rendered endotoxemic by 100 microg/kg lipopolysaccharide (LPS) were examined with measurements of variables reflecting acute lung injury (ALI). ALI induction by LPS was confirmed by blood gas derangement, lung vascular hyperpermeability, and histopathological changes, and was characterized by the deposition of fibrin in the alveolar spaces, bronchioles and vessels. Plasminogen activator inhibitor-1 (PAI-1) and tissue factor (TF) were highly expressed in lungs after LPS injection. While the peaks in levels of PAI-1 and TF were comparable (12 approximately 13-fold from control), their expression time-courses were different: PAI-1 exhibited a bell-shaped expression pattern and peak at 6 h, whereas TF level reached maximum at 10 h. Of note, LPS induced a rapid and significant increase in levels of PAR-1 compared to control, with a peak level at 1 h (3.3-fold). Although declining thereafter, it remained significantly higher than the control level throughout the study period. Expressions of PAR-2, -3, and -4 were also increased by LPS with different time courses from PAR-1 expression. Immunofluorescence staining for PAR-1 were localized in blood vessels, bronchial epithelium, and alveolar pneumocytes after LPS. These results suggest that the increased expression levels of PARs, in addition to PAI-1 and TF, may, in part, underlie the development of ALI occurring during endotoxemia. PMID- 15543323 TI - Ultrasound affects distribution of plasminogen and tissue-type plasminogen activator in whole blood clots in vitro. AB - Ultrasound of 2 MHz frequency and 1.2 W/cm(2) acoustic intensity was applied to examine the effect of sonication on recombinant tissue-type plasminogen activator (rt-PA)-induced thrombolysis as well as on the distribution of plasminogen and t PA within whole blood clots in vitro. Thrombolysis was evaluated quantitatively by measuring clot weight reduction and the level of fibrin degradation product D dimer (FDP-DD) in the supernatant. Weight reduction in the group of clots treated both with ultrasound and rt-PA was 35.2% +/-6.9% which is significantly higher (p<0.0001) than in the group of clots treated with rt-PA only (19.9% +/-4.3%). FDP-DD level in the supernatants of the group treated with ultrasound and rt-PA increased sevenfold compared to the group treated with rt-PA alone, (14895 +/ 2513 ng/ml vs. 2364 +/-725 ng/ml). Localization of fibrinolytic components within the clots was accomplished by using gel-entrapping technique and immunohistochemistry. Spatial distributions of t-PA and plasminogen showed clearly that ultrasound promoted the penetration of rt-PA into thrombi significantly (p<0.0001), and broadened the zone of lysis from 8.9 +/-2.6 microm to 21.2 +/-7.2 microm. We speculate that ultrasound enhances thrombolysis by affecting the distribution of rt-PA within the clot. PMID- 15543324 TI - The factor VII activating protease G511E (Marburg) variant and cardiovascular risk. AB - A previous study had shown a strong relationship between a variant in factor VII activating protease (FSAP G511E) and advanced carotid atheroma. In-vitro, the variant has reduced fibrinolytic but normal pro-coagulant activity, which may constitute a prothrombotic state. The current study has addressed risk for coronary heart disease in a prospective study of cardiovascular disorders (Northwick Park Heart Study II). An interactive effect upon risk was found between the 511E allele and elevated levels of cholesterol and triglyceride. Fibrinogen could substitute for triglyceride levels in this risk-interaction analysis. The findings support the proposal that the FSAP 511E allele exacerbates atherosclerosis or its clinical sequelae. PMID- 15543325 TI - Incidence of post-thrombotic syndrome in patients with previous pulmonary embolism. A retrospective cohort study. AB - There is little information available about the true incidence of post-thrombotic syndrome (PTS) after pulmonary embolism (PE). The aim of this study was to investigate the incidence of PTS in patients with previous pulmonary embolism without concomitant ultrasonographically-detectable deep vein thrombosis (DVT). A retrospective cohort study was conducted at a single tertiary care centre, Cosenza, Italy. Forty-seven consecutive patients with proved PE without DVT within the previous 2 to 6 years, 45 patients with previous DVT in the same years, and 45 patients with diseases unrelated to venous thromboembolism (VTE) underwent a blind assessment for PTS using a clinical score. Two of 47 (4.2%, 95%CI: 0.01-9.9) patients with PE, 2 of 45 (4.4%, 95%CI: 0.01-10.4) patients with diseases unrelated to VTE, and 23 of 45 (53.3%, 95%CI: 38.7-67.9) patients with DVT showed signs and symptoms of PTS. The difference between the first two groups was not statistically significant (p = 0.7). In conclusion, the incidence of PTS after pulmonary embolism without DVT is low, and no different from that of patients without previous VTE. PMID- 15543326 TI - Compensated activation of coagulation in patients with abdominal aortic aneurysm: effects of heparin treatment prior to elective surgery. AB - Elective surgery of abdominal aortic aneurysm (AAA) sometimes leads to excessive bleeding and disseminated intravascular coagulation (DIC), even in patients with normal preoperative coagulation parameters. Coagulation screen, performed routinely before surgery is of limited value in the assessment of compensated activation of the haemostatic system. In this study, we used a number of additional tests (D-dimer, prothrombin fragment 1+2, antithrombin, and activation of fibrinolysis in the platelet poor plasma) for the diagnosis of compensated activation of the haemostatic system in AAA-patients. D-dimer and marker of thrombin generation (prothrombin fragment 1+2) positively correlated with each other (r = 0.768, P < 0.001). Out of 71 AAA patients, 15 patients had normal global coagulation times, but those with a D- dimer concentration above 3000 ng/ml were selected for preoperative low molecular weight heparin (LMWH) treatment. Administration of LMWH diminished coagulation abnormalities (D-dimer and prothrombin fragment 1+2 decreased significantly) and resulted in the increase of platelet number and fibrinogen concentration, indicating their previous consumption. Despite differences in aneurysm diameters between the groups of 15 LMWH treated patients (mean 70.9 +/- 16 mm) and the reference group of 20 untreated AAA patients (mean 52.3 +/- 8.0 mm), intraoperative parameters (operation time, blood loss and transfusion demands) were similar. PMID- 15543327 TI - Effects of epidural-and-general anesthesia combined versus general anesthesia alone on the venous hemodynamics of the lower limb. A randomized study. AB - Our hypothesis was that, due to its sympatholytic action, epidural anesthesia (EA) administered as part of anesthesia in abdominal surgery would generate a marked venous leg flow enhancement, thus aiding in the prevention of peroperative venous stasis. We studied, and comprehensively quantified the venous haemodynamic changes in the lower limb during and immediately after abdominal surgery performed under EA and general (GA) anesthesia combined, in comparison to GA alone. This is a prospective, randomized, controlled study, stratified for hypertension and smoking, comprising ASA 1-2 patients undergoing elective total abdominal hysterectomy. Those with peripheral vascular or chronic venous disease, prior DVT or BMI>35 were excluded. Eligible recruits received either GA (Group GA) (n = 10; age 36-65, median 50) alone or epidural anesthesia (EA) and GA combined (Group EA/GA) (n = 9; age 32-58, median 46). EA (L(1-2)) was administered using lignocaine 2%. Both groups had GA induced with fentanyl and propofol, maintained with N(2)O and isoflurane; larygoscopy was facilitated with vecuronium; analgesia was provided either with morphine (Group GA) or epidurally with 2% lignocaine boli (Group EA/GA). Hemodynamics were determined at the popliteal vein in the horizontal supine position at baseline (resting prior to anesthesia), post epidural (20 min after delivery of EA), post induction (15 min after laryngeal intubation), surgery (upon uterus removal) and recovery (30 min after extubation). There was no difference in the mean velocity[V(mean)] between the 2 groups at baseline (p = 0.35([Mann-Whitney])), and post induction (p = 0.5([Mann-Whitney])). However V(mean) was significantly higher in Group EA/GA than Group GA, both at surgery (point estimate[PE]: 1.8 cm/s; 95% CI: 0.01, 6.3 cm/s; p <0.05([Mann-Whitney])) and recovery (PE: 2.6 cm/s; 95% CI: 0.4, 5.1 cm/s; p = 0.02([Mann-Whitney])). Volume flow[V(Q)] was similar in the 2 groups at baseline and post induction (both, p >0.1([Mann-Whitney])), but was significantly higher in Group EA/GA at surgery (PE: 54 ml/min; 95% CI: 18, 159 ml/min; p = 0.045([Mann-Whitney])) and recovery (PE: 49 ml/min; 95% CI: 16, 129 ml/min; p=0.0037([Mann-Whitney])). Peak velocity, V(mean) and V(Q) increased significantly post epidural in Group EA/GA. Contrary to the venous leg flow attenuation in elective abdominal surgery under GA and upon its recovery, EA administered as part of GA is associated with a significant enhancement of both V(mean) and V(Q). This beneficial hemodynamic effect of EA at the vulnerable stage of recovery may be critically essential in light of enhanced blood viscosity, fibrinolytic shut-down, endothelial/platelet activation and immobility, acting in synergy with putative cardiorespiratory protection. The results of this study lend support to the preferential selection of combined EA/GA in subjects at high risk for venous thromboembolism, particularly when optimal DVT prophylaxis is practically unattainable due to limitations pertaining to the nature of surgery. PMID- 15543328 TI - Warfarin prophylaxis and venous thromboembolism in the first 5 days following hip and knee arthroplasty. AB - Many orthopaedic surgeons use warfarin to prevent venous thromboembolism (VTE) following hip or knee arthroplasty. Since warfarin's antithrombotic effects are delayed, we hypothesized that early VTE (occurring within 5 days post operatively) would be more common in arthroplasty patients receiving warfarin monotherapy compared to those receiving enoxaparin. We performed a secondary analysis of a case-control study examining risk factors for post-operative thrombosis in postmenopausal women. We defined cases as patients who were diagnosed with thrombosis within 5 days of surgery. Controls without thrombosis were matched with cases by age, surgeon, year of surgery and surgical joint. 84 women with early post-operative thrombosis (cases) were matched with 206 controls. 18 cases (21.4%) had been prescribed warfarin mono-therapy, compared with 7 controls (3.4%). 58 (69.1%) cases and 195 (94.7%) controls had been prescribed subcutaneous enoxaparin 30 mg twice daily, starting 12-24 hours after surgery. The odds ratio for any early thrombosis in patients receiving warfarin as opposed to enoxaparin 30 mg twice daily was 8.6 (p<0.0001). For proximal thrombosis, the odds ratio was 11.3 (p<0.0001). Multivariate analysis did not alter these findings. Warfarin's delayed antithrombotic effects may not provide adequate VTE prophylaxis in the immediate post-operative setting. We suggest caution in employing warfarin monotherapy following joint arthroplasty. PMID- 15543329 TI - Over-the-counter vitamin K1-containing multivitamin supplements disrupt warfarin anticoagulation in vitamin K1-depleted patients. A prospective, controlled trial. AB - Most multivitamin supplements contain far less vitamin K(1) than thought to affect warfarin anticoagulation. Having described 3 patients with multivitamin warfarin interactions, we hypothesized that vitamin K(1)-depleted patients are sensitive to even small increments. Therefore, we compared the effect of a vitamin K(1)-containing multivitamin on warfarin anticoagulation between patients with low versus normal vitamin K(1) status. We screened 102 warfarin-treated patients and recruited nine with "low" (< 1.5 mcg/L, 10(th) percentile) (group 1) and 7 with "normal" (>4.5 mcg/L, median) (group 2) total vitamin K(1) plasma levels (vitamin K(1) + vitamin K(1) 2,3-epoxide). Patients received one multivitamin tablet containing 25 mcg of vitamin K(1) daily, for 4 weeks (period 1). A predefined algorithm was used to adjust warfarin doses if the INR was outside the therapeutic range. Patients requiring warfarin increments were then switched to 4 weeks of a vitamin K(1)-free multivitamin supplement (period 2). During period 1, subtherapeutic INRs occurred in 9/9 and 1/7 patients in group 1 and 2, respectively (p <0.001). In group 1, INR decreased by a median of 0.51 (p <0.01), and warfarin dose had to be raised by 5.3% (p <0.01), whereas INR and warfarin dose did not change significantly in group 2. During period 2 (7 patients), there were trends towards decreased total vitamin K(1) and rising INRs associated with significantly lower warfarin doses. We conclude that vitamin K(1) containing multivitamins reduce INR in patients with low vitamin K(1) status. Our study suggests that vitamin K-depleted patients are sensitive to even small changes in vitamin K(1) intake. PMID- 15543330 TI - Thrombin activatable fibrinolysis inhibitor and its fibrinolytic effect in normal pregnancy. AB - We investigated changes in both thrombin activatable fibrinolysis inhibitor (TAFI) antigen levels and its functional effect on in vitro fibrinolysis in normal pregnancy. 152 pregnant women and 31 women in the immediate postpartum period were studied, with pregnancy divided into 6 windows at 4 weekly intervals. As TAFI influences and is in turn influenced by components of the protein C (PC) pathway, its measurements were correlated with levels of soluble thrombomodulin, PC, protein S (PS) and the overall phenotype of activated PC resistance (APCR). Compared with mean TAFI levels at booking gestation (6.6 +/- 1.2 microg/ml), levels peaked at 35-39 weeks gestation (9.6 +/- 2 microg/ml, p = 0.001), followed by a significant drop within 24 hours of delivery (7.2 +/- 1.1 microg/ml). In functional terms, the mean clot lysis time (CLT) (101 +/- 13 min at booking) also peaked at 35-39 weeks gestation (141 +/- 42 min, p = 0.007) and dropped after delivery (99 +/- 33 min), and was significantly correlated with gestational age (r = 0.410, p = 0.001) and could be abrogated in the presence of an inhibitor to TAFI activation. A significant negative correlation was found between TAFI levels and APCR (r = -0.478, p <0.001), APCRV (r = -0.598; p <0.001), PS (r = -0.490, P <0.001) and PC (r = -0.198, p = 0.02). In summary, there is a significant increase in TAFI levels, which translates into increased CLT during pregnancy. Furthermore, changes in TAFI contribute to the increasing APCR of pregnancy. PMID- 15543331 TI - Fibrinogen adsorption and platelet adhesion to metal and carbon coatings. AB - In order to study the haemocompatibility of metal and carbon coatings, fibrinogen adsorption and platelet adhesion to various coatings have been investigated. Two metallic coatings--titanium and zirconium, and two carbon coatings - isotropic diamond-like and isotropic graphite-like coatings, were prepared by plasma vapour deposition onto stainless steel substrate. It has been shown that the adsorption of fibrinogen to metal and carbon coatings and its post-adsorptive transition are dependent on both the material properties and the fibrinogen environment. The adsorption of fibrinogen from human plasma on titanium and zirconium coatings is similar to that on uncoated stainless steel surface. Both carbon coatings adsorb much greater amount of fibrinogen from plasma, and fibrinogen retention by carbon surfaces is also greater than by metal surfaces. Increased numbers of adhered platelets have been found on both carbon coatings in comparison to the metal materials, although this does not correlate with the amount of adsorbed fibrinogen. PMID- 15543332 TI - Glanzmann thrombasthenia Frankfurt I is associated with a point mutation Thr176Ile in the N-terminal region of alpha IIb subunit integrin. AB - In this study, we report on the characterization of a patient with Glanzmann thrombasthenia (GT). Immunochemical analysis on platelets from the patient showed that the expression of alpha IIb beta 3 was only 25% of that in normal healthy controls, suggesting a case of GT. Functional analysis revealed a total lack of fibrinogen binding capacity. Molecular genetic analysis of the full-length cDNA sequences of alpha IIb and beta 3 subunits showed a novel point mutation C621T in alpha IIb cDNA, leading to a missense substitution of threonine for isoleucine at position 176. Coexpression of normal beta 3 and mutant alpha IIb(1176) isoform in mammalian cells showed a marked reduction in the expression of alpha IIb beta 3 heterodimer when compared to the wild-type and a decreased intracellular level of alpha IIb. The T176 I mutation is located in the N-terminal region in the W3:1-2 connecting strand of the beta-propeller. These data suggest that the N-terminal alpha IIb domain plays an important structural role in the formation of heterodimer and that it is also involved in fibrinogen binding. PMID- 15543333 TI - Oxidative status of platelets in normal and thalassemic blood. AB - Chronic platelet activation may be involved in thromboembolic complications, a leading cause of morbidity and mortality in beta-thalassemia. Oxidative stress, with the generation of reactive oxygen species (ROS), is suspected to play a role in the patho-physiology of thalassemia and cardiovascular disorders. In the present study, we adapted flow cytometric techniques to measure oxidative state markers, ROS generation and reduced glutathione (GSH) content in platelets. Our results show that platelets obtained from beta-thalassemic patients contain higher ROS and lower GSH levels than do platelets from normal donors, indicating a state of oxidative stress. In the absence of any known inherent abnormality in thalassemia platelets, this may be attributed to continuous exposure to oxidative insults from extra-platelet sources. We found that exposure of platelets to oxidants such as hydrogen peroxide and tertbutylhydroperoxide or to the platelet activators thrombin, calcium ionophore or phorbol myristate acetate stimulated the platelets' oxidative stress. This was also increased by plasma of thalassemia patients, and decreased following treatment of the plasma with the iron-chelator Desferoxamin. Iron and hemin, the levels of which are augmented in plasma of thalassemia patients, stimulated the platelets' oxidative stress. The oxidative status of the platelets was also affected by red blood cells (RBC); it was higher in normal platelets incubated with thalassemic RBC than with normal RBC. Normal RBC stimulated with hydrogen peroxide had a greater effect on platelets than did unstimulated RBC. The platelets' oxidative stress was ameliorated by antioxidants such as N-acetyl-L-cysteine and vitamin C. Our findings indicate that in thalassemia, platelets undergo a state of oxidative stress, leading to their activation and potentially to thromboembolic consequences, and suggest that this hypercoagulable state might be treated with antioxidants. PMID- 15543334 TI - Association between the Thr715Pro P-selectin gene polymorphism and soluble P selectin levels in a multiethnic population in South London. AB - The aim was to investigate whether the Thr715Pro P-selectin polymorphism is associated with soluble P-selectin (sP-selectin) levels in individuals from different ethnic groups. Plasma sP-selectin and Thr715Pro (A/C) P-selectin gene polymorphism were measured in 237 white (106 females), 177 black African origin (92 females) and 201 South Asian (94 females) individuals living in England. All were free from coronary heart disease (CHD), stroke and other cardiovascular disease, diabetes, drug therapy for hypertension or high lipids, hormone replacement therapy or oral contraceptive pill. The Thr715Pro C allele was rare in blacks (0.8%) and intermediate in South Asians (3.0%) compared to whites (11.2%; p <0.001). sP-selectin levels were significantly lower in the individuals with the AC or CC compared to the AA genotype in both whites (-25% (95% C.I. 33.3 to -16.9); p <0.001) and South Asians (-25.2% (-40.5 to -6.1); p <0.012). There was insufficient power for this analysis in blacks. In conclusion, in whites and South Asians the C allele of the Thr715Pro P-selectin polymorphism is associated with lower sP-selectin levels. Lower levels of sP-selectin were not accounted for by this polymorphism in blacks, in whom the C allele was very rare. PMID- 15543335 TI - Neutrophils stimulated by apolipoprotein(a) generate fragments that are stronger inhibitors of plasmin formation than apo(a). AB - Apolipoprotein(a), the plasminogen-like component of lipoprotein(a), is transformed into fragments by polymorphonuclear neutrophils (PMNs) elastase. Since stimulated PMNs express urokinase-type plasminogen activator (uPA), we sought to investigate the relevance of apo(a) fragmentation on plasminogen activation by neutrophils. Freshly isolated human PMNs stimulated by a 10 kringle recombinant apo(a), r-apo(a), activate plasminogen in a specific and saturable manner (Km = 476 +/- 42 nM, Vmax = 896 +/- 18 pmol min(-1)). This activation is prevented by amiloride, an inhibitor of u-PA, and epsilon-aminocaproic acid, epsilon-ACA, a lysine analogue that blocks plasminogen binding to PMNs. Stimulation of PMNs by apo(a) results in the formation of elastase-derived apo(a) fragments. These fragments produce a concentration-dependent decrease in the formation of plasmin. Addition of elastase inhibitors to PMNs prevented degradation of apo(a) and partially restored the formation of plasmin. In a similar manner, isolated r-apo(a) fragments were able to produce a 100% decrease in plasmin generation as compared to intact r-apo(a). These data indicate that apo(a) fragments produce a more pronounced inhibition in the generation of cell bound plasmin by uPA than the parent apo(a). These effects of apo(a) and its fragments were neutralised by a monoclonal antibody directed against the lysine binding site of apo(a). This mechanism may be of biological relevance to the effects of Lp(a) in conditions where PMNs accumulate and release elastase, i.e. thrombus lysis and inflammatory lesions. PMID- 15543336 TI - Strenuous, acute exercise suppresses polymorphonuclear leukocyte respiratory burst under adherence to surface-adherent platelets in men. AB - Interaction between polymorphonuclear leukocyte (PMN) and platelets is important in the pathogenesis of thrombosis and inflammation. This study investigates how strenuous, acute exercise affects PMN oxidative burst activity under adherence to surface-adherent platelets. Thirty sedentary healthy men exercised strenuously (up to maximal oxygen consumption) on a bicycle ergometer. Before and immediately after exercise, the kinetics of oxidant production, phosphorylation of various protein kinase C (PKC) isoforms, and translocation of p47(phox) in PMNs under adherence to surface-adherent platelets were measured using fluorescence microscopy combined with computerized image analysis. Analytical results can be summarized as follows: (i) either treating the platelet with P-selectin (CD62P) and glycoprotein IIb/IIIa (CD41) antibodies or treating the PMN with beta 2 integrin (CD18) and Mac-1 (CD11b) anti-bodies and PKC zeta pseudosubstrate effectively inhibits platelet-promoted oxidant production of PMN; (ii) PMNs adhesion to surface-adherent platelets is associated with a higher amount of phospho-PKC zeta and a larger ratio of membrane to cytosolic p47(phox) than suspended PMNs; (iii) strenuous, acute exercise decreases platelet-promoted oxidant production of PMN and is accompanied by suppressed phosphorylation of PKC zeta, translocation of p47(phox), and inhibition of PKC zeta pseudosubstrate to oxidant production; (iv) no significant changes occur in PKC alpha/beta II and delta phosphorylation of adherent PMNs following this exercise. Therefore, we conclude that strenuous, acute exercise suppresses platelet-promoted oxidative burst of PMN, possibly by reducing phosphorylation of PKC zeta and translocation of the cytosolic p47(phox) to the plasma membrane, thus inhibiting the assembly and activation of NADPH oxidase in PMN. PMID- 15543337 TI - The effect of homocysteine reduction by B-vitamin supplementation on markers of endothelial dysfunction. AB - Hyperhomocysteinemia is a risk factor for arterial vascular disease and venous thrombosis. The pathophysiology of this relation is unclear, but several studies suggest that hyperhomocysteinemia impairs endothelial function. We examined the effect of homocysteine lowering by B-vitamin supplementation on tissue plasminogen activator (tPA), plasminogen activator inhibitor type 1 (PAI) and von Willebrand factor (vWf)--markers of endothelial dysfunction--in hyperhomocysteinemic and normohomocysteinemic volunteers. A total of 123 healthy volunteers were randomized to placebo or B-vitamins (5 mg folic acid, 0.4 mg hydroxycobalamin and 50 mg pyridoxine) daily for 8 weeks. Before and after the intervention period, blood samples were taken for measurements of homocysteine, tPA, PAI and vWf. There was no evident association between homocysteine concentration and concentrations of markers of endothelial dysfunction at baseline. The mean reduction of homocysteine concentration was 31% (95%CI 22.7 to 39.1) in the B-vitamin group compared to 3% reduction in the placebo group. Concentrations of tPA, PAI and vWf did not change after supplementation of B vitamins. In conclusion, the results of our study show that homocysteine reduction by B-vitamin supplementation has no effect on markers of endothelial dysfunction in healthy volunteers. PMID- 15543338 TI - Amino acid mutagenesis within ligand-binding loops in alpha v confers loss-of function or gain-of-function phenotype on integrin alpha v beta 3. AB - The crystal structure of alpha(v)beta(3) in complex with a cyclic RGD-containing ligand has recently been demonstrated. However, the functional significance of each residue within ligand binding loops has not been fully elucidated. Here, by employing alanine-scanning mutagenesis, we have examined the functional role of ligand contact residues in alpha(v). Tyr178 --> Ala substitution (Tyr178Ala) and Asp218Ala abolished a monovalent ligand, WOW-1 Fab binding as well as soluble fibrinogen binding, which is in perfect agreement with the crystallography. However, Asp150Ala showed no or only a modest inhibition of ligand binding. In contrast, Tyr substitution at Ala215 (Ala215Tyr) increased WOW-1 Fab binding, suggesting that the substitution increased the integrin affinity. The adhesion assay to immobilized fibrinogen showed essentially the same data as obtained using soluble ligands. Our present data indicate that Tyr178 and Asp218, but not Asp150 in alpha(v) is critically involved in ligand-binding and that Ala215 could regulate the affinity of alpha(v)beta(3). PMID- 15543339 TI - The K+-channel opener NS1619 increases endothelial NO-synthesis involving p42/p44 MAP-kinase. AB - Ca(2+)-activated K(+) channels with large conductance (BK(Ca)) have been shown to play an important role in the regulation of vascular tone. We examined the role of the p42/p44 MAP-kinase (p42/p44(MAPK)) on nitric oxide (NO) production in human endothelial cells induced by the BK(Ca)-opener NS1619. Using DiBAC fluorescence imaging a concentration-dependent (2.5-12.5 microM) hyperpolarization induced by NS1619 was observed. A significant increase of intracellular Ca(2+)-concentration by NS1619 was seen using Fura-2-fluorescence imaging, which was blocked by 2-APB, or reduction of extracellular Ca(2+) (n=30; p<0.05). A cGMP-radioimmunoassay was used to examine NO synthesis. NS1619 significantly increased cGMP levels which was inhibited by LNMMA, iberiotoxin, BAPTA, 2-APB, reduction of extracellular Ca(2+), PD 98059, or U0126 (cGMP (pmol/mg protein): NS1619 3.25 +/- 0.85; NS1619 + L-NMMA 0.86 +/- 0.02; NS1619 + iberiotoxin 0.99 +/- 0.09; NS1619 + BAPTA 0.93 +/- 0.29; NS1619 + 2-APB 0.99 +/- 0.31; NS1619 + Ca(2+)-reduction 1.17 +/- 0.06; NS1619 + PD98059 1.06 +/- 0.49; NS1619 + U0126 1.10 +/- 0.24; n=10; p<0.05). The phosphorylation of eNOS and p42/p44(MAPK) was examined by immunocytochemistry. Phosphorylation of p42/p44(MAPK) was significantly increased after 10 minutes of NS1619 stimulation, whereas eNOS phosphorylation was not changed over a period of 1 to 30 minutes. NS1619-induced hyperpolarization was not affected by treatment with PD 98059 or U0126. Additionally, NS1619 inhibited endothelial proliferation involving a NO dependent mechanism. Our data demonstrate that NS1619 causes a transmembrane Ca(2+)-influx leading to an increased NO production involving p42/p44(MAPK). This rise of NO formation is responsible for the NS1619 induced reduction of endothelial cell growth. PMID- 15543340 TI - High plasma heparin cofactor II activity protects from restenosis after femoropopliteal stenting. AB - High heparin cofactor II (HCII) activity has recently been described to protect from coronary instent restenosis, presumably by inactivating thrombin in injured arteries. In this study, we investigated the association of HCII activity and restenosis after femoropopliteal stenting. We studied 63 consecutive patients with peripheral artery disease who underwent femoropopliteal stent implantation after initial failure of plain balloon angioplasty due to a significant residual stenosis (>30% lumen diameter reduction) or a flow limiting dissection. HCII activity was measured before stenting and patients were followed for median 10 months (interquartile range 6 to 17) for the occurrence of a first instent restenosis, defined as a >50% lumen diameter reduction by color coded duplex sonography and confirmed by angiography. Cumulative freedom from restenosis at 6 and 12 months in patients with lower HCII activity (100%, lower tertile, n=20) was 84% and 35% as compared to 93% and 72% in patients with high HCII activity (>100%, middle and upper tertile, n=43; p=0.024 by Log Rank test). Adjusting for the material of the implanted stents (nitinol vs. Wallstents), patients with a high HCII activity had a 0.39-fold reduced risk for instent restenosis (95% CI 0.17 to 0.90, p=0.028), additional adjustment for diabetes mellitus, poor run off, critical limb ischemia and cumulative length of the stented segment did not alter the observed effect. Higher activity of heparin cofactor II may exert a protective effect against instent restenosis also in the femoropopliteal vessel area, confirming a prior observation after coronary stenting. PMID- 15543341 TI - Percutaneous endoluminal arterial cryoenergy improves vascular remodelling after angioplasty. AB - This study aimed to investigate the effect of percutaneous endoluminal arterial cryoenergy immediately after balloon angioplasty on vascular remodeling. Restenosis, the main complication after coronary artery angioplasty, is a complex phenomenon in which vascular remodeling plays a prominent role. Observations of reduced scarring in freeze-induced wounds suggest potential value for cryoenergy in the prevention of restenosis. Juvenile swine underwent a first oversized balloon injury in both carotid arteries (CA) (3 injury sites/artery) and a second injury at day 14. At that time, one CA was randomly assigned to endoluminal cryoenergy of the sequential segments (proximal, medial, distal) at -15 degrees C, -30 degrees C, and -50 degrees C for 120 sec, and the other CA was used as control. Animals were sacrificed 28 days after the second procedure. Compared with intact reference segments, angioplasty reduced both the luminal (LA) and the external elastic lamina (EEL) areas from 6.66+/-0.59 to 3.13+/-0.54 mm(2) (p<0.05) and from 8.81+/-0.81 to 6.48+/-0.52 mm(2) (p<0.05), respectively. Cryoenergy, at the temperature with maximal benefits (-50 degrees C), caused a temperature-related protection, as the LA was maintained (6.79+/-0.89 versus 7.18+/-0.78 mm(2) for reference) and the EEL area increased from 9.12+/-0.78 to 12.98+/-1.07 mm(2), p<0.05. Moreover, cryoenergy increased the density of collagen III (p<0.05) which correlated with the maintenance of the LA (r=0.8045, p<0.009). Cryoenergy prevents late luminal loss after double-injury angioplasty by improving vascular remodeling, and is an interesting new therapeutic approach for the prevention of restenosis after angioplasty. The increased synthesis of collagen III appears to be involved in this phenomenon and could be a potential method of stabilizing the vulnerable plaque. PMID- 15543342 TI - Effect of Interleukin-6 promoter polymorphisms in survivors of myocardial infarction and matched controls in the North and South of Europe. The HIFMECH Study. AB - Elevated plasma IL-6 levels have been implicated in the pathogenesis of coronary heart disease. We have investigated the association of two polymorphisms in the promoter of IL-6 (-572G>C and -174G>C) with levels of inflammatory markers and risk of myocardial infarction (MI) in a European study of MI survivors and age matched controls from two high-risk centres in the North of Europe, and two low risk centres in the South. IL-6 and CRP levels were similar in controls in both regions, but were higher in cases. For the -174G>C polymorphism the rare -174C allele showed a regional difference in allele frequency, being more common in the North European group (0.43 vs 0.28; p < 0.0005), where -174C allele carriers showed an apparent reduced risk of MI compared to -174GG homozygotes (OR 0.53, 95%CI 0.32, 0.86). No such effect was observed in the South or with the -572G>C in either group. Neither genotype was associated with a significant effect on plasma IL-6 levels in either cases or controls. Furthermore, no regional difference was observed in the frequency of the -572G>C SNP, suggesting that these polymorphisms are unlikely to be contributing to the observed increased risk of cardiovascular disease in Northern Europe. PMID- 15543343 TI - Statin and fibrate treatment of combined hyperlipidemia: the effects on some novel risk factors. AB - The effects of cerivastatin and fenofibrate on proteins involved in haemostasis and on markers of inflammation were investigated in otherwise healthy middle-aged males with combined hyperlipidemia. Besides classical risk factors, other so called novel risk factors for coronary artery disease are seen to be playing an increasingly important role in the development and progression of atherosclerosis. Thirty-eight males, aged 49 +/-5 years were randomised to 12 weeks treatment either with cerivastatin at a daily dose of 0.2 mg to 0.4 mg to achieve the LDL cholesterol goal of <3.0 mM, or with fenofibrate 250 mg daily. Fasting serum lipids, homocysteine, total and free tissue factor pathway inhibitor (TFPI), plasminogen activator inhibitor (PAI-1) and tissue plasminogen activator (t-PA) antigen and activity, C-reactive protein (CRP), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured. No change in homocysteine level was observed in the cerivastatin group, while after fenofibrate administration it increased (p <0.0001). Total TFPI decreased significantly after cerivastatin (p = 0.002), but not after fenofibrate. Free TFPI did not decrease after either drug. Neither drug affected (t-PA) antigen and activity, while fenofibrate increased PAI-1 antigen (p <0.05) and activity (p <0.05). Cerivastatin decreased serum CRP values by 49.5% (p = 0.001), and fenofibrate by 29.8% (p = 0.03). The decreases of CRP in the two groups differed significantly (p = 0.04). IL-6 levels decreased significantly in the fenofibrate group (39%; p <0.0001), but not in the cerivastatin group (15%; p = 0.24) No significant decreases were observed for TNF-alpha. Cerivastatin had neutral effects on fibrinolysis, homocysteine or coagulation. On the other hand, fenofibrate increased PAI-1 antigen and activity and homocysteine, and did not affect coagulation. Both cerivastatin and fenofibrate reduced CRP levels, the decrease being significantly greater after cerivastatin. Fenofibrate also significantly decreased IL-6. PMID- 15543344 TI - An in vitro study on the mechanisms of coagulation activation in acute myelogenous leukemia (AML): role of tissue factor regulation by cytotoxic drugs and GM-CSF. AB - AML patients may suffer from a disseminated coagulopathy, which can aggravate a pre-existing bleeding tendency due to thrombocytopenia and platelet dysfunction. The cellular and molecular mechanisms underlying this coagulopathy, however, are not completely understood. Indeed, the broad and increasing therapeutic use of cytotoxic drugs and growth factors is likely to contribute to the complexity of hemostatic abnormalities encountered in this hematologic malignancy. The nature of coagulation activation in AML was therefore investigated in vitro using the human leukemic cell line, HL60. Tissue factor (TF) was almost entirely located on the cell surface and bound factor VIIa, but only 15-25% of this TF was primarily functionally active. Treatment with increasing concentrations of daunorubicin or cytosine-beta-D-arabinofuranoside, two cytotoxic drugs commonly used in AML therapy, induced apoptosis and secondary necrosis of HL60 cells and resulted in marked decryption of TF PCA independent of de novo protein synthesis. This PCA modulating effect was concomitant with and functionally dependent on the exposure of phosphatidylserine on the outer membrane leaflet. Similar observations were made in analogous ex vivo studies on patient-derived myeloblasts. Incubation of HL60 cells with GM-CSF, a cytokine expressed in the bone marrow microenvironment and used as an adjunct to AML treatment, evoked a cellular response, which included both enhanced TF production and release of VEGF-A and uPA into the culture medium. We conclude that both decryption of pre-formed TF PCA by chemotherapeutic drugs and de novo induction of TF by cytokines such as GM-CSF can regulate the pro-coagulant phenotype of HL60 cells in vitro. PMID- 15543345 TI - Familial pernicious anaemia with hyperhomocysteinaemia in recurrent early pregnancy loss. PMID- 15543346 TI - B cell depletion may lead to normalization of anti-platelet, anti-erythrocyte and antiphospholipid antibodies in systemic lupus erythematosus. PMID- 15543347 TI - Percutaneous coronary intervention with stenting in a patient with haemophilia A and an acute myocardial infarction following a single dose of desmopressin. PMID- 15543348 TI - Liver transplantation for type 1 primary hyperoxaluria as a cure for combined thrombophilia. PMID- 15543349 TI - Morphological evidence that 2-arachidonoylglycerol is a true agonist of human platelets. PMID- 15543350 TI - Two new factor X mutations (Pro382Leu and Phe356Cys) associated with low activity and low antigen levels. PMID- 15543351 TI - Plasminogen activator inhibitor-1 plasma level increases with age in subjects with the 4G allele at position -675 in the promoter region. PMID- 15543352 TI - A survey of thrombosis prophylaxis use in patients with lower limb fractures. PMID- 15543353 TI - Local and general factors are the likely cause of venous thrombosis in lower limb arthroplasty. PMID- 15543354 TI - [The importance of the clinico-radiological correlation in the diagnosis of congenital heart disease]. PMID- 15543355 TI - [Unsupervised or semi supervised cardiac rehabilitation. A practical alternative]. PMID- 15543356 TI - [Renin-angiotensin-aldosterone system genetic polymorphisms associated with coronary artery disease and its anatomic features]. PMID- 15543357 TI - Endomyocardial biopsy foretells ventricular function recovery after coronary artery bypass grafting. AB - OBJECTIVE: Patients with ischemic heart failure may benefit from coronary artery bypass grafting. The histopathological variables associated with improvement in ejection fraction 6 months after surgery were assessed. METHODS: This study comprised 24 patients indicated for coronary artery bypass grafting, ejection fraction < 35%, functional class II-IV heart failure, and mean age 59 +/- 9 years. Endomyocardial biopsies were performed during and 6 months after surgery. Extension of the fibrosis, number of cells with myocytolysis, and hypertrophy of the muscle fiber were quantified by using a system of image analysis. Clinical and functional review was repeated within 6 months. RESULTS: A significant improvement in heart failure functional class was observed in 16 patients after 6 months of follow-up (from NYHA functional class 2.8 +/- 0.7 to 1.7 +/- 0.6; P < 0.001), but the ejection fraction did not change (25 +/- 6 % vs. 26 +/- 10%). Hypertrophy of the muscle fiber was similar in the specimens biopsied in the pre- and postoperative periods (21 +/- 4 vs. 22 +/- 4 microm), but the extension of fibrosis (8 +/- 8 vs. 21 +/- 15% area) and the number of cells with myocytolysis (9 +/- 11 vs. 21 +/- 15% cell) significantly increased. However, the composition of a histological score combining those 3 variables indicated a greater increase in the ventricular function of those with a lower degree of preoperative histopathological alterations. CONCLUSION: Patients with ischemic cardiomyopathy undergoing coronary artery bypass grafting improved their ventricular function when the preoperative adverse histopathological alterations were of a lower degree. PMID- 15543358 TI - Acute effects of the use of estrogens in association with progestogens on postprandial triglyceridemia and vascular reactivity. AB - OBJECTIVE: To assess whether hormone replacement therapy with estrogens in association with progestogens in postmenopausal hypertensive women alters postprandial triglyceridemia and vascular reactivity. METHODS: A double-blind, placebo-controlled, crossover study was carried out with 15 postmenopausal women (age range: 50 to 70 years, mean = 61.6 +/- 6 years) randomly assigned to 2 weeks of placebo or oral ingestion of 0.625 mg of equine conjugated estrogens and 2.5 mg of medroxyprogesterone, fed a high-fat diet (897 calories; 50.1% fat). Vascular reactivity (VR - % of vessel diameter variation in the fasting period and 2 hours after meals) was measured by using the automated ultrasound method. Lipid profile and glycemia during the fasting period and 2 hours after a high-fat meal were measured. RESULTS: With placebo, vascular reactivity (VR) decreased from 3.20 +/- 17% during the fasting period to -2.1 +/- 30% 2 hours after the meal (P = 0.041). With the hormone replacement therapy, vascular reactivity decreased from 6.14 +/- 27% during the fasting period to - 0.05 +/- 18% 2 hours after the meal (P = NS). Postprandial triglyceridemia increased as follows: 35 +/ 25% with placebo; and 12 +/- 10% with hormone replacement therapy (P < 0.05). CONCLUSION: In postmenopausal hypertensive women, 2 weeks of hormone replacement with an association of estrogens and progestogens decreased hypertriglyceridemia after a high-fat meal, an effect that may reduce the endothelial dysfunction occurring in the postprandial period. PMID- 15543359 TI - Guide of internet sites for the study of cardiology. AB - OBJECTIVE: To increase the offer of didactic resources for medical students, physicians, and health care professionals, stimulating the use of the Internet for academic purposes or cardiological updating. METHOD: Electronic addresses with academic content in the areas of anatomy, biophysics, physiology, semiology, electrocardiography, and image diagnosis were researched and selected. The selection criteria included the following: relevance of the content, neatness of presentation, and richness of animation resources. The sites obtained were classified in regard to their contents and academic level. RESULTS: The sites obtained were as follows: 5 sites of anatomy and anatomical pathology, one of biophysics, 3 of physiology, 8 of semiology, 7 of image diagnosis, and 2 of electrocardiography. The sites were also organized according to academic level to provide an alternative access. The set of addresses resulted in a simplified and hierarchic guide of contents for the study of cardiac morphology and image diagnosis in cardiology. CONCLUSION: The resulting list of sites is an example of the potential of the Internet as a learning instrument to be used in association with other conventional pedagogical methods. PMID- 15543360 TI - The A1166C polymorphism of the angiotensin II type-1 receptor in acute myocardial infarction. AB - OBJECTIVE: To assess the association of the A1166C polymorphism of the angiotensin II type-1 receptor (AT1R) gene with acute myocardial infarction and also with the severity of coronary artery disease. METHODS: A prospective, cross sectional study was carried out with 110 patients with acute myocardial infarction, who, on coronary angiography, had significant lesions (> 50%) assessed according to 3 criteria of severity: number of vessels affected, morphology of the atherosclerotic plaque, and coronary risk score. The control group comprised 104 individuals with no coronary lesions. The A1166C polymorphism of AT1R gene was determined by polymerase chain reaction in the DNA of leukocytes in peripheral blood. The classic coronary risk factors were analyzed in all individuals. RESULTS: When stratifying the genotypes in regard to risk factors, only smoking predominated in the AC heterozygous patients (P = 0.02). The genotypic frequency in the infarcted patients was as follows: AA = 54.5%; AC = 35.5%; and CC = 10%, which was similar and nonsignificant in regard to that in the control group (P = 0.83). No risk increase occurred for acute myocardial infarction when comparing the genotypes as follows: CC vs AA (OR = 1.35; 95% CI = 0.50 - 3.59); AC vs AA (OR = 1.03; 95% CI = 0.58 - 1.84); and AA+AC vs AA (OR = 1.33; 95% CI = 0.51 - 3.45). None of the severity criteria showed a significant correlation with the genotypes. CONCLUSION: According to our results, no correlation exists between the A1166C polymorphism of the angiotensin II type-1 receptor (AT1R) gene and acute myocardial infarction or the severity of coronary artery disease. PMID- 15543361 TI - Myocardial contrast echocardiography in patients with suspected or known coronary artery disease: comparison with myocardial nuclear scintigraphy. AB - OBJECTIVE: To compare myocardial contrast echocardiography (MCE) using PESDA and adenosine in bolus (ADN) with myocardial nuclear scintigraphy (NS) in patients (pts) undergoing routine investigation with a high probability of having coronary artery disease. METHODS: This study comprised 125 pts (85 men) with 58.4 +/- 10.6 years, who underwent MCE and NS within 4 weeks. MCE was performed with PESDA in a continuous infusion at rest and after administration of an adenosine bolus. The LV walls was divided into 3 territories related to the coronary arteries, in a total of 375 territories. MCE was normal when an increase in contrast intensity occurred after ADN. The reduction in contrast intensity at rest or after ADN was defined as an abnormal MCE result. NS was performed according to classical protocols. When compared per patient, both examinations were considered concordant when they were normal or abnormal, independent of its location. The comparison by territory was considered concordant when perfusion defects existed or not in the same territory. The chi-square test was used to determine the significance of concordance. RESULTS: In 106/125 pts, MCE and NS were concordant (84.8% - P < 0.001). Concordance occurred in 342/375 territories (91.2% - P < 0.001). For the LAD territory, concordance was 87.2%; for the RCA, 93.6%; and for the CX, 92.8% (P < 0.001). CONCLUSION: An excellent concordance exists between MCE and NS in assessing pts for coronary artery disease; therefore, MCE may represent a good alternative for assessing myocardial perfusion. PMID- 15543362 TI - Prevalence, awareness, and control of systemic arterial hypertension in the state of Rio Grande do Sul. AB - OBJECTIVE: To report the prevalence of cardiovascular risk factors, particularly of systemic arterial hypertension, in the adult population of the Brazilian state of Rio Grande do Sul, in addition to the public's level of awareness, hypertensive control, and associated factors. METHODS: A cross-sectional, population-based study with random sampling from a population pool was carried out with 918 patients older than 20 years from 1999 to 2000. Systemic arterial hypertension was defined as blood pressure > or = 140/90 or current use of antihypertensive drugs. RESULTS: The prevalence of systemic arterial hypertension was 33.7% (n = 309), and 49.2% of the individuals were unaware of their hypertensive condition; 10.4% knew they were hypertensive, but did not undergo treatment; 30.1% underwent antihypertensive treatment, but did not have adequate control; and 10.4% underwent antihypertensive treatment with adequate control. Based on multivariate analysis, the following variables were found to be significantly associated with the presence of systemic arterial hypertension: age (OR = 1.06), obesity (OR = 3.03), and low educational level (OR = 1.82). These same variables were associated with unawareness of the hypertensive condition: age (OR = 1.05), obesity (OR = 2.46), and low educational level (OR = 2.17). CONCLUSION: Prevalence of systemic arterial hypertension in the state of Rio Grande do Sul has been remained at constant levels for the past decades, and the population's level of awareness of it has improved slightly. However, the control level of systemic arterial hypertension has not increased. This study allowed the definition of a target group -- elderly, obese individuals with low educational level -- for both diagnostic campaigns and better control of blood pressure levels. PMID- 15543363 TI - Syncope and complete atrioventricular block related to pulmonary thromboembolism. PMID- 15543364 TI - Stent implantation in critical stenosis of the celiac trunk: enlarging the frontiers of percutaneous vascular intervention. PMID- 15543365 TI - [Standardization of equipment and technics for supervised cardiovascular rehabilitation]. PMID- 15543366 TI - Screening for abdominal aortic aneurysm. PMID- 15543367 TI - Hemodynamics and renal function during administration of low-dose dopamine in severely ill patients. AB - CONTEXT: Although a large number of studies have been performed regarding the renal and hemodynamic effects of the infusion of low-dose dopamine (LDD) in severely ill patients, there is still controversy on this subject. OBJECTIVE: To evaluate the effects of dopamine (2 microg/kg/min) on systemic hemodynamics (lowest mean arterial pressure, MAP, highest heart rate, HR, central venous pressure, CVP), creatinine clearance (CLcr), diuresis and fractional sodium excretion (FENa+). TYPE OF STUDY: A non-randomized, open, prospective clinical trial. SETTING: An intensive care unit in a tertiary university hospital. PARTICIPANTS: 22 patients with hemodynamic stability admitted to the intensive care unit. PROCEDURES: Patients were submitted to three two-hour periods: without dopamine (P1), with dopamine (P2) and without dopamine (P3). MAIN MEASUREMENTS: The above mentioned variables were measured during each period. CLcr was assessed based upon the formula U x V/P, where U is urinary creatinine (mg/dl), V is diuresis in ml/min and P is serum creatinine (mg/dl). FENa+ was calculated based upon the formula: urinary sodium (mEq/l) x P/plasma sodium (mEq/l) x U) x 100. Results were presented as mean and standard deviation. The Student t test was used and results were considered significant if p was less than 0.05. RESULTS: Twelve patients (seven males and five females) were included, with a mean age of 55.45 years. There was no significant variation in MAP, HR, CVP, CLcr or FENa+ with a dopamine dose of 2 microg/kg/min. On the other hand, diuresis significantly increased during P2, from 225.4 to 333.9 ml. CONCLUSION: Infusion of 2 microg/kg/min of dopamine for 2 hours increases diuresis. At the doses studied, dopamine does not induce significant alterations in MAP, HR, CVP, CLcr and FENa+. PMID- 15543368 TI - Totally implantable venous catheters for chemotherapy: experience in 500 patients. AB - CONTEXT: Totally implantable devices are increasingly being utilized for chemotherapy treatment of oncological patients, although few studies have been done in our environment to analyze the results obtained from the implantation and utilization of such catheters. OBJECTIVE: To study the results obtained from the implantation of totally implantable catheters in patients submitted to chemotherapy. TYPE OF STUDY: Prospective. SETTING: Hospital do Cancer A.C. Camargo, Sao Paulo, Brazil. METHODS: 519 totally implantable catheters were placed in 500 patients submitted to chemotherapy, with preference for the use of the right external jugular vein. Evaluations were made of the early and late stage complications and patient evolution until removal of the device, death or the end of the treatment. RESULTS: The prospective analysis showed an average duration of 353 days for the catheters. There were 427 (82.2%) catheters with no complications. Among the early complications observed, there were 15 pathway hematomas, 8 cases of thrombophlebitis of the distal stump of the external jugular vein and one case of pocket infection. Among the late-stage complications observed, there were 43 infectious complications (0.23/1000 days of catheter use), 11 obstructions (0.06/1000 days of catheter use) and 14 cases of deep vein thrombosis (0.07/1000 days of catheter use). Removal of 101 catheters was performed: 35 due to complications and 66 upon terminating the treatment. A total of 240 patients died while the catheter was functioning and 178 patients are still making use of the catheter. CONCLUSION: The low rate of complications obtained in this study confirms the safety and convenience of the use of totally implantable accesses in patients undergoing prolonged chemotherapy regimes. PMID- 15543369 TI - Clinical and demographic profile of users of a mental health system for medical residents and other health professionals undergoing training at the Universidade Federal de Sao Paulo. AB - CONTEXT: A postgraduate and resident trainee mental health assistance center was created in September 1996 within our university. OBJECTIVE: To describe the clinical and demographic profile of its users. TYPE OF STUDY: Retrospective. SETTING: Universidade Federal de Sao Paulo--Escola Paulista de Medicina (Unifesp EPM). METHODS: The study was carried between September 1996 and November 2002, when 233 semi-structured registration forms were filled out either by the psychologist or the psychiatrist during their first contact with the trainees, who were medical and nursing residents, and postgraduate students at specialization, master or doctoral levels. The registration forms included demographic, occupational and clinical data. RESULTS: The trainees were predominantly young (mean of 27 years old), single (82.0% of cases), women (79.4%), seeking help especially during the first year of training (63.1%). In 70.8% of the cases, they came to the service spontaneously. Such individuals showed greater adherence to the treatment than those who were referred by supervisors (p < 0.05). In 30% of the cases, the trainee sought psychological guidance or support at the service due to specific situational conflicts. Depression and anxiety disorders were the most frequent diagnoses; 22.3% of the trainees followed up mentioned a tendency towards suicidal thoughts. In comparison with other trainees, there was a higher prevalence of males among the medical residents (p < 0.01), with more cases of sleep disorders (p < 0.05), a smaller number of individuals refraining from the use of alcohol (p < 0.05) and a higher number of trainees requiring leave of absence (p < 0.001). DISCUSSION: The first year of training in health sciences is the most stressful, especially for women. Depression and anxiety symptoms are common, reflecting transitory self limited deadaptation. However, the severity of the cases can also be evaluated in view of the large number of trainees who mentioned suicidal tendencies. CONCLUSIONS: This study emphasizes the need and importance of providing formal, structured and confidential mental health services for medical residents and postgraduate students from other health professions, in the training programs of academic institutions. PMID- 15543370 TI - Prevalence of abdominal aortic aneurysms--a screening program in Sao Paulo, Brazil. AB - CONTEXT: Abdominal aortic aneurysm is an asymptomatic but potentially fatal condition. Elective surgery can prevent death from rupture, and is indicated for aneurysms larger than 45 mm. Because aneurysms tend to grow with time, detection of small ones (> 29 mm) may lead to a closer follow-up of patients at risk. OBJECTIVE: To determine the prevalence of abdominal aortic aneurysms in Sao Paulo, Brazil. DESIGN: Prospective, descriptive. SETTING: University Hospital. PARTICIPANTS: Persons aged 50 years or more were offered, through the press, the opportunity to be screened for abdominal aortic aneurysm. The total number screened was 2,756. PROCEDURE: All were submitted to abdominal palpation and ultrasound examination. PARAMETER STUDIED: A maximum diameter of 30 mm or more was considered to be an aneurysm. RESULTS: Sixty-four aneurysms were detected, nine of which measuring more than 49 mm. Palpation detected 60 aneurysms, but only 20 of these were confirmed by the ultrasound. Conversely, 41 of the ultrasound-detected aneurysms were not palpable. The percentages of abdominal aortic aneurysms found in the subgroups via ultrasound examination (with 95% confidence interval) were as follows: total group, 2.3 (1.8-3); men, 4.6 (3.5 5.9); women, 0.6 (0.3-1.1); men aged 60 or more, 6 (4.3-8); women aged 60 or more, 0.9 (0.4-1.8). CONCLUSION: In Sao Paulo, Brazil, 1.8 to 3 % of persons aged 50 years or more are expected to have abdominal aortic aneurysms. In the subgroup of men aged 60 or more, the expected prevalence is between 4.3 and 8%. PMID- 15543371 TI - Reassessment of diagnostic criteria in cutaneous lymphocytic infiltrates. AB - CONTEXT: Non-specific lymphocytic infiltrates of the skin pose difficulties in daily practice in pathology. There is still a lack of pathognomonic signs for the differential diagnosis between benign and malignant lymphocytic infiltrates. OBJECTIVE: To evaluate the morphological and immunohistochemical profile of lymphocytic infiltrations of the skin according to clinical outcome. TYPE OF STUDY: Retrospective; histopathological and immunohistochemical analysis. SETTING: Referral center, university hospital. SAMPLE: 28 cases of lymphocytic infiltrates of difficult differential diagnosis selected from the records. MAIN MEASUREMENTS: Eighteen histological variables and the immunophenotypic profile were assessed using the CD4, CD8, CD3, CD20 and CD30 lymphoid markers and compared to subsequent follow-up. RESULTS: The most common diagnoses were: initial mycosis fungoides (eight cases) and drug reactions (five cases). Single morphological variables did not discriminate between benign and malignant infiltrates except for the presence of Pautrier-Darier's microabscesses, which were found only in mycosis fungoides (p = 0.015). Patterns of superficial and deep infiltration (p = 0.037) and also the presence of eosinophils (p = 0.0207) were more frequently found in benign lymphocytic infiltrates. Immunohistochemical profile of T-cell subsets showed overlap between benign and malignant infiltrates with a predominance of CD4-positive (helper) lymphocytes in the majority of cases. CONCLUSIONS: A combination of clinical and histological features remains the most reliable approach for establishing a definite diagnosis in cases of lymphoid skin infiltrates. PMID- 15543372 TI - Multiple drug resistance protein (MDR-1), multidrug resistance-related protein (MRP) and lung resistance protein (LRP) gene expression in childhood acute lymphoblastic leukemia. AB - CONTEXT: Despite the advances in the cure rate for acute lymphoblastic leukemia, approximately 25% of affected children suffer relapses. Expression of genes for the multiple drug resistance protein (MDR-1), multidrug resistance-related protein (MRP), and lung resistance protein (LRP) may confer the phenotype of resistance to the treatment of neoplasias. OBJECTIVE: To analyze the expression of the MDR-1, MRP and LRP genes in children with a diagnosis of acute lymphoblastic leukemia via the semiquantitative reverse transcription polymerase chain reaction (RT-PCR), and to determine the correlation between expression and event-free survival and clinical and laboratory variables. DESIGN: A retrospective clinical study. SETTING: Laboratory of Pediatric Oncology, Department of Pediatrics, Faculdade de Medicina de Ribeirao Preto, Universidade de Sao Paulo, Brazil. METHODS: Bone marrow aspirates from 30 children with a diagnosis of acute lymphoblastic leukemia were assessed for the expression of messenger RNA for the MDR-1, MRP and LRP genes by semi-quantitative RT-PCR. RESULTS: In the three groups studied, only the increased expression of LRP was related to worsened event-free survival (p = 0.005). The presence of the common acute lymphoblastic leukemia antigen (CALLA) was correlated with increased LRP expression (p = 0.009) and increased risk of relapse or death (p = 0.05). The relative risk of relapse or death was six times higher among children with high LRP expression upon diagnosis (p = 0.05), as confirmed by multivariate analysis of the three genes studied (p = 0.035). DISCUSSION: Cell resistance to drugs is a determinant of the response to chemotherapy and its detection via RT-PCR may be of clinical importance. CONCLUSIONS: Evaluation of the expression of genes for resistance to antineoplastic drugs in childhood acute lymphoblastic leukemia upon diagnosis, and particularly the expression of the LRP gene, may be of clinical relevance, and should be the object of prospective studies. PMID- 15543373 TI - Radiofrequency fistulotomy: a better alternative for treating low anal fistula. AB - CONTEXT: Wide varieties of approaches are employed in dealing with low anal fistula. However, the simple method of laying open the fistula tract (fistulotomy) is still considered to be the favored one. MATERIALS AND METHODS: A modified approach to the procedure of fistulotomy is discussed. This study describes the procedure, which used a technique of radiofrequency surgery, and its outcome in 232 patients with low anal fistula. The patients were followed for a period of 15 months. RESULTS: The patients were discharged on the same day as the procedure. The mean period off work was four days. The average healing time recorded was 67 days. Four wound complications in the form of premature closure of the external wound were noted, which required trimming of the edges. Two of these wounds remained unhealed. The recurrence rate was 1.7%. CONCLUSION: In this era when the emphasis is on criteria like the minimization of hospital stay, reduction of postoperative pain, early resumption of work and low and comparable recurrence rates, there is a future for the procedure of radiofrequency fistulotomy. PMID- 15543374 TI - Phospholipid transfer protein activity in two cholestatic patients. AB - CONTEXT: Plasma phospholipid transfer protein mediates the transfer of phospholipids from triglyceride-rich lipoproteins, very low density lipoproteins and low density lipoproteins to high density lipoproteins, a process that is also efficient between high density lipoprotein particles. It promotes a net movement of phospholipids, thereby generating small lipid-poor apolipoprotein AI that contains particles and subfractions that are good acceptors for cell cholesterol efflux. CASE REPORT: We measured the activity of plasma phospholipid transfer protein in two cholestatic patients, assuming that changes in activity would occur in serum that was positive for lipoprotein X. Both patients presented severe hypercholesterolemia, high levels of low density lipoprotein cholesterol and, in one case, low levels of high density lipoprotein cholesterol and high levels of phospholipid serum. The phospholipid transfer activity was close to the lower limit of the reference interval. To our knowledge, this is the first time such results have been presented. We propose that phospholipid transfer protein activity becomes reduced under cholestasis conditions because of changes in the chemical composition of high density lipoproteins, such as an increase in phospholipids content. Also, lipoprotein X, which is rich in phospholipids, could compete with high density lipoproteins as a substrate for phospholipid transfer protein. PMID- 15543375 TI - Insulin autoimmune syndrome: case report. AB - CONTEXT: Insulin autoimmune syndrome (IAS, Hirata disease) is a rare cause of hypoglycemia in Western countries. It is characterized by hypoglycemic episodes, elevated insulin levels, and positive insulin antibodies. Our objective is to report a case of IAS identified in South America. CASE REPORT: A 56-year-old Caucasian male patient started presenting neuroglycopenic symptoms during hospitalization due to severe trauma. Biochemical evaluation confirmed hypoglycemia and abnormally high levels of insulin. Conventional imaging examinations were negative for pancreatic tumor. Insulin antibodies were above the normal range. Clinical remission of the episodes was not achieved with verapamil and steroids. Thus, a subtotal pancreatectomy was performed due to the lack of response to conservative treatment and because immunosuppressants were contraindicated due to bacteremia. Histopathological examination revealed diffuse hypertrophy of beta cells. The patient continues to have high insulin levels but is almost free of hypoglycemic episodes. PMID- 15543376 TI - Pan-cytokeratin immunoexpression in Wilms' tumors: a simple approach for understanding tumor epithelial differentiation. AB - Wilms' tumor is one of the most common solid tumors in children and is an interesting model for understanding the pathogenesis of embryonal tumors. Cytokeratins are intracellular fibrous proteins present in tissue of epithelial origin. The immunoexpression of the pan-cytokeratin AE1AE3 was studied in paraffin-embedded tissue sections from 24 Wilms' tumors (12 with nephrogenic rests) and also tissue samples from 15 corresponding normal kidneys, to evaluate epithelial differentiation in the genesis of Wilms' tumor. We observed that the intensity of the expression of AE1AE3 in the epithelial component of Wilms' tumors directly correlated with the degree of maturity of the epithelial structures correspondent to the collecting ducts. PMID- 15543377 TI - Dysplasia in the Barrett's esophagus: utopia, difficult or impossible concordance. PMID- 15543378 TI - [Dysplasia in Barrett's esophagus--intra- and interobserver variability in histopathological diagnosis]. AB - BACKGROUND: Barrett's esophagus is a well-known pre-malignant condition. Pathologic interpretation of biopsy specimens guides endoscopic surveillance as well as the therapeutic approach that will be carried out. However, the predictive value of histopathologic diagnosis can be questioned due to its poor intra- and interobserver reproducibility. AIMS: To assess intra- and interobserver variability in the diagnosis of Barrett's dysplasia. MATERIAL AND METHODS: Three-micrometer thick sections from biopsy specimens from 42 patients with Barrett's esophagus were stained with hematoxylin-eosin and PAS-alcian blue. The reading of the slides was carried out blindly in a light microscope. Intra and interobserver variability in the interpretation of the slides was determined by kappa statistics. RESULTS: The number of tissue specimens was 229, with average of 5.45 (1 to 18) fragments for patient. Low grade dysplasia was diagnosed by pathologists in 21.4% to 52.4% of the cases. The intra-observer agreement for the diagnosis of low grade dysplasia was slight (kappa = 0.30). The interobserver agreement for the diagnosis of low grade dysplasia was poor, with kappa scores between 0.05 and 0.16. The diagnosis of dysplasia, with agreement for all pathologists examining the same set of slides, was 14.3%. CONCLUSIONS: Pathologic interpretation of Barrett's dysplasia may be subject to marked intra- and interobserver variabiliaty. Interpretation of low grade dysplasia, as high grade dysplasia, should also be considered for review by two or more pathologists. PMID- 15543379 TI - Vertical transmission of hepatitis C virus in a hospital in southern Brazil. AB - BACKGROUND: There still are controversies concerning the vertical transmission of hepatitis C virus. AIM: To evaluate the prevalence of antibodies against hepatitis C virus in pregnant women, as well as the rate of vertical transmission of this virus. PATIENTS AND METHODS: Between August 1998 and November 1999, 1,090 consecutive pregnant women were screened for anti-hepatitis C virus; positive results were confirmed by the polymerase chain reaction assay. Patient's viral load was evaluated by the branched deoxyribonucleic acid assay. Hepatitis C virus genotype was identified by direct sequencing of the polymerase chain reaction amplification products. The same tests were performed in the children born from infected mothers at the 1st and 6th month of life. RESULTS: Of the 1,090 mothers surveyed, 29 were positive for anti-hepatitis C virus (prevalence of 2.66%). Twenty-five patients presented with hepatitis C virus RNA, with a median hepatitis C virus viral load of 3.132 +/- 5.891 MEq/mL. Twenty-two patients (six human immunodeficiency virus-coinfected) were followed and gave birth to 23 children; 18 of them had blood samples tested at the 1st month of life, and 22, at the 6th month. Vertical transmission rate was 5.56%; it affected a girl who had hepatitis C virus RNA detectable only in the 1st month sample (41.570 MEq/mL). The mother who transmitted hepatitis C virus was coinfected with human immunodeficiency virus and presented with an hepatitis C virus viral load of 3.765 MEq/mL, with 100% homology with her daughter's hepatitis C virus genotype. CONCLUSION: These results suggest that the prevalence of hepatitis C virus infection in pregnant women should not be neglected, and early diagnosis of vertical transmission and the follow up of infected children should be emphasized. PMID- 15543380 TI - Determination of carcinoembryonic antigen levels in peripheral and draining venous blood in patients with colorectal carcinoma. AB - BACKGROUND: The problem of the relationship between blood carcinoembryonic antigen (CEA) levels and tissue CEA content in colorectal carcinoma, and the mechanisms for CEA release from tumor cells in tissue adjacent to the neoplasm is important to understanding the biology of colorectal carcinoma. It has not been adequately explained whether CEA in the peripheral blood is drained mainly by portal system blood or by the lymphatic system, or indeed by both systems. AIM: To study the behavior of CEA levels in peripheral blood (CEA-p) and venous effluent blood (CEA-d) among patients with colorectal tumors, who underwent curative operation. METHOD: A total of 28 patients were studied (12 male [42.9%] and 16 female [57.1%], mean age 66.1 years [range: 43 - 84]). Immediately after laparotomy, peripheral venous blood was extracted by antecubital venous puncture and venous effluent blood was collected from the main drainage vein of the lesions. Values of CEA-p, CEA-d and the gradient between CEA-d and CEA-p that were less than 5.0 ng/mL were considered normal. RESULTS: Eight (28.6%) patients were stage A in Duke's classification, nine (32.1%) stage B and 11 (39.3%) stage C. The neoplasm was located in the rectum of 14 patients (50.0%), in the transverse colon in five (17.9%), in the sigmoid in four (14.3%), in the cecum and/or ascending colon in three (10.7%), and in the descending colon in two (7.1%). The histopathological examination revealed well-differentiated adenocarcinoma in all the patients. Only one patient (3.6%), Duke's classification stage C, presented neoplasm with venous invasion. The gradient between the CEA-p and CEA-d levels were normal in 25 patients (88.3%) and high in three (10.7%). The mean value for CEA-p was 3.8 +/- 4.1 ng/mL (0.1-21.1 ng/mL) and for the drained CEA (CEA-d) it was 4.5 +/- 4.3 ng/mL (0.3-20.2 ng/mL), without significant difference between these values. There was a significant difference between the mean value for CEA-p and CEA-d levels greater than 5 ng/mL. CONCLUSION: The CEA-p and CEA-d levels in the colorectal carcinoma patients were not shown to be different. The results from this study suggest that, in colorectal neoplasm without venous invasion, there may not be notable CEA drainage from the tumor by the portal vein effluent blood. PMID- 15543381 TI - [Sensibility of anthropometric-laboratory markers of protein-energy malnutrition in cirrhotic patients]. AB - AIM: The prevalence and severity of protein-energy malnutrition (PEM) were investigated through the anthropometric (body mass index, triceps and subescapular skinfolds and upper arm circumferences) and blood measures in 117 cirrhotic patients. The sensitivity and specificity of single or combined PEM markers were tested among Child A (n=18), Child B (n=42) and Child C (n=57) adults (51+/-13y). RESULTS: Were calculated as z score and considered deficient when z< or =-1.28SD according to local standards. The most deficient markers where albumin (93%), hemoglobin (90%), upper arm circumference (61%), triceps (55%) and subescapular (53%) skinfolds. By combining upper arm circumference with triceps or subescapular skinfolds, PEM were detected in 63% of patients varying from 39-44% (Child A) to 64-68% (Child B or C). CONCLUSION: Thus the pattern of PEM present in cirrhosis is predominantly in their protein compartment and worsened with the severity of hepatocellular insuficiency. Upper arm circumference can be used as sensitive markers of presence and severity of PEM in cirrhotic patients but showing low specificity for discriminate PEM among Child grades (B and C) of hepatocellular dysfunctions. PMID- 15543382 TI - The influence of endoscopic procedures upon the contamination of Helicobacter pylori cultures. AB - BACKGROUND: Among the various diagnostic methods for the detection of Helicobacter pylori infection, histological examination and microbiological processing of gastric biopsy samples are assumed to be the gold standard techniques. AIMS: Since H. pylori culture can be affected by the presence of non H. pylori bacteria, we evaluated the efficacy of endoscope disinfection and the influence of endoscopic procedures on culture contamination. PATIENTS AND METHODS: The procedures used during the first two routine endoscopies were evaluated during 28 consecutive days. Endoscopy room, forceps and endoscopic channel were analyzed before and after the beginning of normal procedures. After disinfection, a biopsy simulation was performed to verify the gastric bacteria. RESULTS: Endoscope disinfection removed all organisms from forceps and endoscopic channel with 100% efficacy. The most frequent non-H. pylori bacteria detected were Streptococcus bovis, Enterobacter hormaechei, and Staphylococcus aureus. The sensibility of the H. pylori culture was affected by the presence of non-H. pylori bacteria. CONCLUSION: The risk of transmission of microorganisms was not detectable when sterilized biopsy forceps and stringent disinfection standards were employed. Whilst S. bovis and E. hormaechei may be common in gastric microbial flora, the presence of P. aeruginosa and S. aureus indicated that the manipulation of biopsies could be responsible for culture contamination with these bacteria. PMID- 15543383 TI - Inflammatory fibroid polyp: an immunohistochemical study. AB - BACKGROUND: Inflammatory fibroid polyp is a localized lesion, which arises in the submucosa of the gastrointestinal tract, most often in the stomach. Although it is generally believed to represent a reactive, nonneoplastic condition, its histogenesis remains controversial. AIM: To study inflammatory fibroid polyp by immunohistochemistry in an attempt to further clarify their histogenesis. MATERIAL AND METHODS: Nine cases were studied by immunohistochemistry using a panel of antibodies against smooth-muscle actin, vimentin, S-100 protein, factor VIII- R.Ag and macrophage (HAM-56). RESULTS: There was a strong diffuse positive staining pattern in the spindle cells with vimentin antibody. A patchy staining for smooth-muscle actin was observed in these cells. Immunophenotyping revealed a heterogeneous reaction with HAM-56. In edematous areas, HAM-56-positive cells show voluminous cytoplasm and reniform nuclei. In cell-rich areas, the HAM-56 positive cells had fusiform cytoplasm. Stains for S-100 and factor VIII RAg were negative in the proliferating elements. CONCLUSIONS: The present immunohistochemical study refutes the suggested neural or vascular nature of the lesion. The strong positivity for vimentin in all cases suggests a major component of spindle cells best recognizable as fibroblasts. These results would favor the existence of a span of morphological and immunohistochemical patterns possibly indicating evolutive phases of an inflammatory reaction. PMID- 15543384 TI - [Gastrointestinal alterations in diabetes mellitus: oxidative stress and blood flow in the superior mesenteric artery--experimental study]. AB - BACKGROUND: Gastrointestinal symptoms are frequent in diabetes mellitus and may be related to the oxidative stress, which is defined by an imbalance between the pro-oxidant and antioxidant systems. AIMS: To assess some of the gastrointestinal changes in the diabetes mellitus model, such as the oxidative stress in the stomach and liver of diabetic animals and the blood flow in the superior mesenteric artery at different times of study. MATERIAL AND METHODS: Male Wistar rats weighing 250-350 g were used and ascribed to four different groups as follows: group I--7 days of diabetes, group II--30 days of diabetes, group III- 60 days of diabetes, and group IV--90 days of diabetes. Diabetes was obtained by administering streptozotocin (70 mg/kg) intraperitoneally. RESULTS: A significant increase in liver and stomach lipoperoxidation of diabetic animals was found only at 90 days. A significant decrease in the activity of antioxidant enzymes catalase e glutathione transferase was found in stomach, whereas in liver only glutathione transferase was significantly decreased. The blood flow in the superior mesenteric artery of animals with 90 days of diabetes was increased as compared to controls. CONCLUSIONS: It is possible to suppose that the increase in the oxidative stress in stomach and liver as well as the change in the blood flow in the superior mesenteric artery are affected by the length of diabetes and the hyperglycemia found in the animals studied, which would determine gastrointestinal changes. PMID- 15543385 TI - [Cytokine participation in the acute rejection of intestinal transplantation in rats]. AB - BACKGROUND: Intestinal transplantation is a possible treatment for patients with short bowel syndrome, aiming the reintroduction of oral diet. However, the major obstacle in this procedure is the strong rejection. Delay in rejection diagnosis may be irreversible and lethal. AIM: To define method for early diagnosis of rejection based on the presence of interleucin-6 (IL-6) e interferon- gamma (IFN gamma) from intestinal allograft. MATERIAL AND METHODS: Isogenic rats Brown Norway (BN) and Lewis (LEW) were submitted to intestinal heterotopic allotransplantation and divided in two groups: LEW donor to LEW recipient isograft group (C) and BN donor to LEW recipient allograft group (Tx). According to the day of sacrifice, Tx group were subdivided in three subgroups with eight animals each as follow: Tx3--sacrificed at third postoperative day (POD), Tx5- sacrificed at fifth POD and Tx7--sacrificed at seventh POD. Eight animals from control group were subdivided in three moments according to the time of biopsy from the graft as follow: C3--biopsy at third POD; C5--biopsy at fifth POD and C7 -biopsy at seventh POD. All animals from control group were sacrificed at seventh POD. Rejection parameters were compared between the control groups (C3 vs C5, C3 vs C7 and C5 vs C7, and allograft group (Tx3 vs Tx5, Tx3 vs Tx7 and Tx5 vs Tx7). The same parameters were analyzed between the control group and allograft groups (C3 vs Tx3, C5 vs Tx5 and C7 vs Tx7). RESULTS: In C group no statistical significant difference regarding the immunoexpression of the cytokines, while in Tx group, immunoexpression of IL-6 and IFN-gamma were remarkable since the fifth postoperative day. PMID- 15543386 TI - [Genetics and immunopathogenics aspects of the celiac disease: a recent vision]. AB - BACKGROUND: Celiac disease, or gluten-sensitive enteropathy, is a strongly inherited condition. Although the genetic association of CD with the DQ2 and DQ8 HLA haplotypes has been known for long, others HLA and non-HLA genes are also important in the development of the disease. Celiac disease results of the combined effect of different normally functioning genes' products. The tissue damage in celiac disease is immunologically mediated and several effector mechanisms are responsible for the disease expression. The interplay between genetic, immunological and environmental factors explains the large spectrum of clinical, histological and serological alterations observed in the different stages of the disease development, pointing out to the polygenic nature of celiac disease. CONCLUSION: The recent advances in the understanding of the immunopathogenesis, genetics and diagnoses of celiac disease have allowed the revision of strict concepts and previous criteria and their adequation to the new evidences, aiming a better diagnostic and orientation to celiac patients and relatives. PMID- 15543387 TI - [Metastatic cutaneous adenocarcinoma of unknown primary site. Case report]. AB - BACKGROUND: Metastases may be the first manifestation of adenocarcinoma. Up to 60% are cutaneous and present in advanced stage neoplasms. Research for the primary site is costly and requires endoscopy, imaging and immunohistochemical exams. The primary site becomes obvious in only 15% to 20% of live patients and is detected mainly at autopsy. AIM: To report a case of metastatic cutaneous moderately differentiated adenocarcinoma of unknown primary site, located in the lower left abdomen. RESULTS: The lesion was surgically resected. Primary site was not found by any imaging or endoscopy exams. The immunohistochemistry was negative for CEA, CK20, PSA and positive for CK7. Based on these exams, prostate and colorectal cancer were excluded. The pancreas and biliary tract were considered as the probable primary site. CONCLUSION: The research for the primary site still is difficult, expensive and is not effective to the treatment of metastatic adenocarcinomas. PMID- 15543388 TI - Celiac disease onset after pegylated interferon and ribavirin treatment of chronic hepatitis C. AB - AIM: Report of a case of a woman patient who developed celiac disease after pegylated interferon alpha-2a and ribavirin use for chronic hepatitis C. PATIENT AND METHOD: A 34-year-old woman with chronic hepatitis C, genotype 3, receiving pegylated interferon alpha-2a and ribavirin for 6 months, developed progressive malaise and anemia 6 months after the end of treatment. RESULT: Additional investigation revealed duodenal villous atrophy and positivity for anti endomysium and anti-gliadin antibodies. Celiac disease diagnosis was performed and symptoms and laboratory abnormalities improved after gluten-free diet. CONCLUSION: Celiac disease must be ruled out in patients with malabsorption complaints in or after interferon (or pegylated interferon) therapy. Screening for celiac disease with detection of anti-endomysium antibodies would be done in susceptible patients. PMID- 15543389 TI - [Recidivation of bilateral longitudinal peritoneum-aponeurotic transposition on incisional hernioplasty]. AB - OBJECTIVES: To carry out a study of the recidivation rate of incisional hernioplasty by means of bilateral longitudinal peritoneum-aponeurotic transposition. METHOD: A retrospective study was carried out in patients from the University Hospital in Belo Horizonte, MG, Brazil and from a surgeon's private clinic who had undergone incisional hernioplasty by means of bilateral peritoneum aponeurotic transposition. A total of 132 patients (80 from University Hospital and 52 from private clinic) were monitored over an average period of 4 years and 10 months. Most the patients were obese and all of them had disloged longitudinal hernias of medium and large volume. Three aspects must be stressed: the procedure for reaching the internal abdominal wall, the posterior aponeurotic incision and the threads used. RESULTS: Recidivation occurred in 7.69% of the patients belonging to a group operated on by one surgeon and in 18.75% of patients operated on by a trainee surgeon. PMID- 15543390 TI - [Quality of life in patients with inflammatory bowel diseases: translation to Portuguese language and validation of the "Inflammatory Bowel Disease Questionnaire" (IBDQ)]. AB - BACKGROUND: Crohn's disease comprises, together with ulcerative colitis, the inflammatory bowel diseases, which because their morbidity and chronic evolution have a considerable impact on the patient's quality of life. The measurement of quality of life requires the determination of the degree of subjective well-being attributed to the lack of symptoms, to psychological state and daily activity, and is useful as an instrument of evaluation, both in clinical trials and health programs. Inflammatory Bowel Disease Questionnaire is an instrument developed by North-American investigators of McMaster University which reproducibility and validity has been determined in several countries studies, and has shown its use in the measurement of the quality of life of patients with inflammatory bowel diseases. AIMS: Our purpose was to translate that instrument into Portuguese language, its adaptation to Brazilian culture and verify its psychometric properties (validity and reproducibility). METHODS: It was applied to a population of 50 inflammatory bowel diseases patients (24 with Crohn's disease, 26 with ulcerative colitis). To test the criteria and construction validity of the instrument's, it was compared with SF-36, a general questionnaire for establishing quality of life, and the Crohn's Disease Activity Index and Lichtiger's Index. RESULTS: The correlations obtained were mostly moderate. About the intestinal diseases activity index, Lichtiger's index showed better correlations than Crohn's Disease Activity Index. To evaluate its reproducibility, the instrument was applied on three occasions, twice by the same observer (12 days average between applications) and once by a second observer, resulting in a high degree of intra- and interobserver correlation. CONCLUSIONS: Portuguese version of Inflammatory Bowel Disease Questionnaire is a valid and reproducible instrument which can be used to evaluate the quality of life of Brazilian inflammatory bowel diseases patients. PMID- 15543392 TI - Low coronary perfusion pressure is associated with endocardial fibrosis in a rat model of volume overload cardiac hypertrophy. AB - Left ventricular hypertrophy following volume overload is regarded as an example of cardiac remodeling without increased fibrosis accumulation. However, infarction is associated with increased fibrosis within the noninfarcted, hypertrophied myocardium, particularly in the subendocardial regions. It is conceivable to suppose that, as also occurs postinfarction, low coronary driving pressure may also interfere with accumulation of myocardial fibrosis following aortocaval fistula. PURPOSE: To investigate the role of acute hemodynamic changes in subsequent deposition of cardiac fibrosis in response to aortocaval fistula. METHOD: Aortocaval fistula were created in 4 groups of Wistar rats that were followed over 4 and 8 weeks: aortocaval fistula 4 and aortocaval fistula 8 (10 rats each) and their respective controls (sham-operated controls - Sh), Sh4 and Sh8 (8 rats each). Hemodynamic measurements were performed 1 week after surgery. Hypertrophy and fibrosis were quantified by myocyte diameter and collagen volume fraction at the end of follow up. RESULT: Compared with Sh4 and Sh8, pulse pressure, left ventricular end-diastolic pressure, and +dP/dt were higher in aortocaval fistula 4 and aortocaval fistula 8, but -dP/dt was similar. Coronary driving pressure (mm Hg), used as an estimate of perfusion pressure, was lower in aortocaval fistula 8 (52.6 +/- 4.1) than in Sh8 (100.8 +/- 1.3), but comparable between aortocaval fistula 4 (50.0 +/- 8.9) and Sh4 (84.8 +/- 2.3). Myocyte diameter was greater in aortocaval fistula 8, whereas interstitial and subendocardial fibrosis were greater in aortocaval fistula 4 and aortocaval fistula 8. Coronary driving pressure correlated inversely and independently with subendocardial fibrosis (r(2) = .86, P <.001), whereas left ventricular systolic pressure (r(2) = 0.73, P = .004) and end-diastolic pressure (r(2) = 0.55, P = 012) correlated positively and independently with interstitial fibrosis. CONCLUSION: Coronary driving pressure falls and ventricular pressures increase early after aortocaval fistula and are associated with subsequent myocardial fibrosis deposition. PMID- 15543393 TI - Oropharyngeal colonization by Haemophilus influenzae in healthy children from Taubate (Sao Paulo), prior to the Haemophilus influenzae type B vaccination program in Brazil. AB - Haemophilus influenzae is one of the most important bacterial agents of otitis and sinusitis. H. influenzae type b (Hib) is one of the main causes of meningitis, pneumonia, and septicemia in nonvaccinated children under 6 years of age. The aims of this study were to determine the prevalence of H. influenzae and Hib oropharyngeal colonization prior to the onset of the Hib vaccination program in Brazil in previously healthy children and to assess the susceptibility profile of this microorganism to a selected group of antimicrobials that are used to treat acute respiratory infections. METHOD: Cultures of Haemophilus influenzae were made from oropharynx swabs from 987 children under 6 years of age who were enrolled in 29 day-care centers in Taubate (a city of Sao Paulo state, Brazil) between July and December 1998. RESULTS: The prevalence of H. influenzae carriers was 17.4%, and only 5.5% of the strains were beta-lactamase producers. The prevalence of Hib carriers was high, 7.3% on average (range, 0.0 - 33.3%). CONCLUSIONS: The low prevalence of colonization by penicillin-resistant strains indicates that it is not necessary to substitute ampicilin or amoxicilin to effectively treat otitis and sinusitis caused by H. influenzae in Taubate. PMID- 15543394 TI - Glycemia in newborns of hypertensive mothers according to maternal treatment. AB - PURPOSE: To evaluate the evolution of glycemic levels in newborns of hypertensive mothers according to maternal treatment. METHODS: Prospective randomized study, including 93 newborns of mothers treated with isradipine (n = 39), atenolol (n = 40), or low sodium diet (control group - n=14). Glycemia was determined at birth (mother and newborn by the oxidase glucose method) and in the 1st, 3rd, 6th, 12th, and 24th hours after birth (newborn by a test strip method). The evolution of glycemia was analyzed in each group (Friedman test). The groups were compared regarding glycemia (Kruskall-Wallis test), and linear regression models were constructed for the analyses (independent variable = maternal glycemia; dependent variables = umbilical cord, 3rd, and 6th hour glycemia). RESULTS: There were no statistically significant differences among the mean blood glucose levels of the 3 groups in any of the assessments. There was a correlation between maternal and umbilical cord blood glucose in the isradipine (r = 0.61; P <.05) and control (r = 0.84; P <.05) groups. Regarding glycemia levels of the mothers and newborns in the third and sixth hours postpartum, this correlation was present only in the control group (maternal x third hour: r = 0.65; P <.05; maternal x sixth hour: r = 0.68; P <.05). There were no correlations in the atenolol group. Hypoglycemia was detected in 51.3% of the isradipine group, 60% of the atenolol group, and 35.7% of the control group, and it was more frequent in the first hour postpartum in all groups. CONCLUSIONS: The results suggest a similar effect of the 3 types of treatment upon newborn glycemia. The correlation analysis suggests that isradipine could have effects upon newborn glycemia only after birth (correlation only in umbilical cord blood), whereas atenolol could act earlier (there was no correlation at any moment). The results also point to the need for glycemic control from the first hour postpartum of newborns of hypertensive mothers whether they have or have not undergone treatment with antihypertensive drugs. PMID- 15543395 TI - Endemic pemphigus foliaceus (fogo selvagem) and pemphigus vulgaris: immunoglobulin G heterogeneity detected by indirect immunofluorescence. AB - Pemphigus are autoimmune intraepidermal blistering diseases in which immunoglobulin G (IgG) autoantibodies are directed against desmosomal glycoproteins. The aim of this study was to determine the IgG subclass profile of endemic pemphigus foliaceus (fogo selvagem) and pemphigus vulgaris utilizing indirect immunofluorescence. PATIENTS AND METHODS: Twenty-five patients with pemphigus vulgaris, 25 with endemic pemphigus foliaceus (fogo selvagem), and 25 healthy controls were analyzed by indirect immunofluorescence for circulating autoantibodies (total IgG and its subclasses). RESULTS: Our data revealed a significant correlation (P <.05) of disease activity and autoantibody levels in both forms of pemphigus, i.e., negative titers related to clinical remission, whereas positive results related to active disease. Immunoglobulin G subclass analysis in fogo selvagem demonstrated that in patients in remission, 56% showed positive immunoglobulin G4; in active disease, immunoglobulin G4 was the predominant subclass (100% positive in all cases). The IgG subclass profile in pemphigus vulgaris showed that in patients in remission, only 10% were positive for immunoglobulin G4; in active disease, positivity for immunoglobulin G4 was present in 78% to 88% of the cases. CONCLUSION: Subclass characterization of immunoglobulin G autoantibodies is a useful tool for pemphigus follow-up, since immunoglobulin G4 (IgG4) is the subclass that is closely related to recognition of pathogenic epitopes, and consequently with disease activity. Careful monitoring should be performed for fogo selvagem in clinical remission with a homogeneous IgG4 response, since this may indicate more frequent relapses. PMID- 15543396 TI - Induction of experimental mammary carcinogenesis in rats with 7,12 dimethylbenz(a)anthracene. AB - PURPOSE: To test an experimental model of chemical mammary carcinogenesis induction in rats. METHODS: Twenty young virgin Sprague-Dawley female rats, aged 47 days, received 20 mg of 7,12-dimethylbenz(a)anthracene (DMBA) intragastrically by gavage. Afterwards, at 8 and 13 weeks, their mammary glands were examined. At the end of the experiment, the animals were sacrificed, and the mammary tumors were measured and weighed. Tumor fragments were analyzed using light microscopy. RESULTS: Eight weeks after DMBA injection, 16 rats presented at least 1 breast tumor (80%). After 13 weeks, all of them (100%) developed breast carcinomas that were confirmed by histopathological analysis. CONCLUSION: This experimental animal model of chemical mammary induced carcinogenesis is feasible and can be used in further experiments on the role of tumorigenic biomodulator substances. PMID- 15543397 TI - Urethral catheter removal 7 or 14 days after radical retropubic prostatectomy: clinical implications and complications in a randomized study. AB - PURPOSE: To evaluate the hypothesis that a 7-day period of indwelling catheter after radical retropubic prostatectomy is effective and safe without the need of performing cystography. METHODS: In the period from January of 2000 to July of 2002, 73 patients underwent radical retropubic prostatectomy, and these patients were prospectively randomized in 2 groups: Group 1-37 patients who had the urethral catheter removed 7 days after the procedure, and Group 2-36 patients who had the catheter removed 14 days after the surgery. The 2 groups were similar, the surgeons and the technique were the same, and no cystography was performed to evaluate the presence of leaks. RESULTS: Two patients in Group 1 had bleeding and clot retention after having the catheter taken out in the seventh postoperative day and were managed by putting the catheter back in for 7 more days. Two patients in Group 2 developed bladder neck stricture and were treated by bladder neck incision with success. The continence rate was the same, with 2 cases of incontinence in each group. About 2 pads a day were used by the patients with incontinence. The average follow-up was 17.5 months (12-36 months). No urinary fistula, urinoma, or pelvic abscesses developed after catheter removal. Two patients were excluded from the analysis of this series: 1 died with a pulmonary embolus in the third postoperative day, and 1 developed a urinary suprapubic fistula before catheter withdrawal, which was maintained for 16 days. CONCLUSION: Withdrawal of the urethral catheter 7 days after radical retropubic prostatectomy, without performing cystography, has a low rate of short-term complications that are equivalent to withdrawal 14 days after the surgery. PMID- 15543398 TI - Renal and urinary findings in 20 patients with Williams-Beuren syndrome diagnosed by fluorescence in situ hybridization (FISH). AB - PURPOSE: Williams-Beuren syndrome is a rare multiple anomalies/mental retardation syndrome caused by deletion of contiguous genes at chromosome region 7q11.23. The aim of this work was to determine the frequency and the types of renal and urinary tract anomalies in 20 patients with Williams-Beuren syndrome. METHODS: The fluorescence in situ hybridization test using a LSI Williams syndrome region DNA probe was performed for all 20 patients to confirm the diagnosis of Williams Beuren syndrome. A prospective study was performed in order to investigate renal and urinary aspects using laboratory assays to check renal function, ultrasonography of the kidneys and urinary tract, voiding cystourethrogram and urodynamics. RESULTS: Deletion of the elastin gene (positive fluorescence in situ hybridization test) was found in 17 out of 20 patients. Renal alterations were diagnosed in 5 of 17 (29%) the patients with the deletion and in 1 of 3 patients without the deletion. Fourteen patients with the deletion presented dysfunctional voiding. Arterial hypertension was diagnosed in 3 patients with deletions and 1 of these presented bilateral stenosis of the renal arteries. CONCLUSIONS: Due to the high incidence of renal and urinary abnormalities in Williams-Beuren syndrome, performing a systematic laboratory and sonographic evaluation of the patients is recommended. PMID- 15543399 TI - Influence of nephrotic state on the infectious profile in childhood idiopathic nephrotic syndrome. AB - Patients with idiopathic nephrotic syndrome present alterations in their cellular and humoral immune reactions that predispose them to the development of infectious processes. PURPOSE: To characterize the infectious processes in patients with idiopathic nephrotic syndrome. PATIENTS AND METHODS: Ninety-two children and adolescents with idiopathic nephrotic syndrome were assessed retrospectively. The types of infection were grouped as follows: upper respiratory tract infections; pneumonia; skin infections; peritonitis; diarrhea; urinary tract infection ; herpes virus; and others. The patients were divided into 2 groups: Group I (steroid-responsive) n = 75, with 4 subgroups-IA (single episode) n = 10, IB (infrequent relapsers) n = 5, IC (frequent relapsers) n = 14, and ID (steroid-dependent) n = 46; and Group II (steroid-resistant) n = 17. The incidence-density of infection among the patients was assessed throughout the follow-up period. Comparisons for each group and subgroup were done during the periods of negative and nephrotic proteinuria. RESULTS: The analysis revealed a greater incidence-density of infections during the period of nephrotic proteinuria in all the groups and subgroups, with the exception of subgroup IA. During the period of nephrotic proteinuria, subgroups IC, ID, and Group II presented a greater incidence-density of infections as compared to subgroup IA. For the period of negative proteinuria, there was no difference in the incidence density of infections between the groups and subgroups. Upper respiratory tract infections were the most frequent infectious processes. CONCLUSION: The nephrotic condition, whether as part of a course of frequent relapses, steroid dependence, or steroid resistance, conferred greater susceptibility to infection among the patients with idiopathic nephrotic syndrome. The results of this study suggest that the best preventive action against infection in this disease is to control the nephrotic state. PMID- 15543400 TI - Microsatellite instability in solitary and sporadic gastric cancer. AB - Recently, the presence of microsatellite instability (MSI) has been reported in gastric cancer and associated with older age of presentation, distal tumor location, early disease staging, and better overall prognosis. Different characteristics in presentation and in tumor behavior may be explained by different genetic alterations during carcinogenesis of gastric cancer. Identification of specific genetic pathways in gastric cancer may have direct impact on prognosis and selection of treatment strategies. PATIENTS AND METHODS: All 24 patients were treated by radical surgery. Fragments of normal and tumor tissues were extracted from the specimen and stored at -80 degrees C before DNA purification and extraction. PCR amplification utilizing microsatellite markers was performed. Tumors presenting PCR products of abnormal sizes were considered positive for microsatellite instability (MSI+). RESULTS: Five patients (21%) had tumors that were MSI+ in at least 1 marker. In the group of patients with Lauren's intestinal-type gastric carcinoma, 3 had tumors that were MSI+ (23%), while in the group of diffuse-type gastric cancer, 2 patients had tumors that were MSI+ (19%). The mean age of presentation and the male:female ratio was similar in both groups. Tumors that were MSI+ were more frequently located in proximal portion of the stomach compared to microsatellite-stable (MSS) tumors (40% vs. 16%). Although there was a trend of patients with MSI+ tumors towards a proximal gastric tumor location, early staging, and negative lymph node metastasis, there was no statistical significance compared to those with MSS tumors (P >.1). Comparison of overall and disease-free survival between gastric tumors that were MSI+ and those that were MSS found no statistically significant differences (P >.1). CONCLUSIONS: Microsatellite instability is a frequent event in gastric carcinogenesis and shows a trend towards distinct clinical and pathological characteristics of gastric cancer. PMID- 15543401 TI - New method for evaluation of cutaneous sensibility in diabetic feet: preliminary report. AB - Diabetic neuropathy is an important complication of the disease, responsible for ulceration and amputation of the foot. Prevention of these problems is difficult mainly because there is no method to correctly access sensibility on the skin of the foot. The introduction of the Pressure-Specified Sensory Device (PSSD) in the last decade made possible the measurement of pressure thresholds sensed by the patient, such as touch, both static and in movement, on a continuous scale. This paper is the first in Brazil to report the use of this device to measure cutaneous sensibility in 3 areas of the foot: the hallux pulp, the calcaneus, and the dorsum, which are territories of the tibial and fibular nerves. METHOD: Non diabetic patients were measured as controls, and 2 groups of diabetic patients - with and without ulcers - were compared. The PSSD was used to test the 3 areas described above. The following were evaluated: 1 PS (1-point static), 1 PD (1 point dynamic), 2 PS (2-points static), 2 PD (2-points dynamic). RESULTS: The diabetic group had poorer sensibility compared to controls and diabetics with ulcers had poorer sensibility when compared to diabetics without ulcers. The differences were statistically significant (P <.001). CONCLUSION: Due to the small number of patients compared, the results should be taken as a preliminary report. PMID- 15543402 TI - Comparison of catheter-related infection risk in two different long-term venous devices in adult hematology-oncology patients. AB - PURPOSE: Infection is the leading complication of long-term central venous catheters, and its incidence may vary according to catheter type. The objective of this study was to compare the frequency and probability of infection between two types of long-term intravenous devices. METHODS: Retrospective study in 96 onco-hematology patients with partially implanted catheters (n = 55) or completely implanted ones (n = 42). Demographic data and catheter care were similar in both groups. Infection incidence and infection-free survival were used for the comparison of the two devices. RESULTS: In a median follow-up time of 210 days, the catheter-related infection incidence was 0.2102/100 catheter-days for the partially implanted devices and 0.0045/100 catheter-days for the completely implanted devices; the infection incidence rate was 46.7 (CI 95% = 6.2 to 348.8). The 1-year first infection-free survival ratio was 45% versus 97%, and the 1-year removal due to infection-free survival ratio was 42% versus 97% for partially and totally implanted catheters, respectively (P <.001 for both comparisons). CONCLUSION: In the present study, the infection risk was lower in completely implanted devices than in partially implanted ones. PMID- 15543403 TI - Anorectal leiomyomas: report of two cases with different anatomical patterns and literature review. AB - Gastrointestinal mesenchymal tumors comprise a rare group of gastrointestinal tract wall tumors that have long been a source of confusion and controversy, especially in terms of pathological classification, preoperative diagnosis, management strategies, and prognosis. This report describes the clinical manifestations and management of 2 rectal leiomyomas and reviews the pertinent literature. Case 1: A 44-year-old woman was admitted reporting a nodule in the right para-anal region for the previous 2 years. At proctological examination, a 4-cm diameter fibrous mass situated in the para-anal region that produced an arch under the smooth muscle on the right rectal wall just above the anorectal ring was noted. Computed tomography and magnetic resonance imaging of the abdomen and pelvis showed the lesion and detected no other abnormalities. Surgical treatment consisted of wide local resection of the tumor through a para-anal incision, with no attempts to perform lymphadenectomy. Case 2: A 40-year-old male patient was admitted reporting constant anal pain for 4 months. He presented a 3-cm submucosal nodule at the anterior rectal wall just above the dentate line. After 2 inconclusive preoperative biopsies, transanal resection of the tumor was performed. Histological analysis of the specimen showed a benign leiomyoma. A review of the literature is presented, emphasizing some clinical and therapeutic aspects of this unusual rectal tumor. PMID- 15543404 TI - The effect of intravenous zoledronic acid on glucocorticoid-induced multiple vertebral fractures in juvenile systemic lupus erythematosus. AB - Glucocorticoids are widely used in the treatment of lupus patients, and adverse effects, which include osteoporosis and associated fractures, are frequent. Treatment of osteoporosis of young patients should be effective and not harmful to bone growth and remodeling. Bisphosphonates are drugs that decrease the incidence of bone fractures, but their use in juvenile patients is still controversial because of their possible side effects on the growing skeleton. However, recently published studies showed that linear growth continued normally after treatment with these drugs, and there was no excessive suppression of bone remodeling or mineralization defects. Zoledronic acid is a new intravenous bisphosphonate that has been approved by the US FDA for use with hypercalcemia of malignancies and might be an effective treatment for postmenopausal osteoporosis. The authors report a case of a young girl with systemic lupus who developed multiple vertebral collapses due to glucocorticoid therapy, and zoledronic acid was used producing significant clinical and densitometric improvement. PMID- 15543405 TI - Hypophosphatemia in critically ill children. AB - The purpose of this paper is to review clinical studies on hypophosphatemia in pediatric intensive care unit patients with a view to verifying prevalence and risk factors associated with this disorder. We searched the computerized bibliographic databases Medline, Embase, Cochrane Library, and LILACS to identify eligible studies. Search terms included critically ill, pediatric intensive care, trauma, sepsis, infectious diseases, malnutrition, inflammatory response, surgery, starvation, respiratory failure, diuretic, steroid, antiacid therapy, mechanical ventilation. The search period covered those clinical trials published from January 1990 to January 2004. Studies concerning endocrinological disorders, genetic syndromes, rickets, renal diseases, anorexia nervosa, alcohol abuse, and prematurity were not included in this review. Out of 27 studies retrieved, only 8 involved pediatric patients, and most of these were case reports. One clinical trial and one retrospective study were identified. The prevalence of hypophosphatemia exceeded 50%. The commonly associated factors in most patients with hypophosphatemia were refeeding syndrome, malnutrition, sepsis, trauma, and diuretic and steroid therapy. Given the high prevalence, clinical manifestations, and multiple risk factors, the early identification of this disorder in critically ill children is crucial for adequate replacement therapy and also to avoid complications. PMID- 15543406 TI - The best infertility treatment for vasectomized men: assisted reproduction or vasectomy reversal? AB - In men with prior vasectomy, microsurgical reconstruction of the reproductive tract is more cost-effective than sperm retrieval with in vitro fertilization and intracytoplasmic sperm injection if the obstructive interval is less than 15 years and no female fertility risk factors are present. If epididymal obstruction is detected or advanced female age is present, the decision to use either microsurgical reconstruction or sperm retrieval with in vitro fertilization and intracytoplasmic sperm injection should be individualized. Sperm retrieval with in vitro fertilization and intracytoplasmic sperm injection is preferred to surgical treatment when female factors requiring in vitro fertilization are present or when the chance for success with sperm retrieval and intracytoplasmic sperm injection exceeds the chance for success with surgical treatment. PMID- 15543407 TI - Blackflies (Diptera: Simuliidae) of Southern Guyana with keys for the identification of adults and pupae--a review. AB - A revision is made of the previously poorly studied blackfly fauna from the south western border of Guyana with Brazil. Notes on the biosystematics of the species found are provided, together with keys and illustrations based on their morphology. Of the 14 species recorded, eight are anthropophilic and two of these (Simulium oyapockense s.l. and S. guianense s.l.) are proven vectors of human onchocerciasis in the nearby Amazonia focus of the disease in neighbouring Brazil. PMID- 15543408 TI - Isoenzimatic analysis of four Anopheles (Kerteszia) cruzii (Diptera: Culicidae) populations of Brazil. AB - Anopheles cruzii is a small sylvatic mosquito and primary human Plasmodium vector in Southern Brazil. The distribution of this bromeliad-breeding mosquito follows the Atlantic forest coastal distribution, where bromeliads are abundant. Morphological, genetic, and molecular polymorphisms among different populations have been reported and it has recently been suggested that An. cruzii is a complex of cryptic species. The aim of this work is to analyze the gene flow between different populations of An. cruzii collected in four localities within the geographic distribution range of the species, and to examine if An. cruzii is a complex of cryptic species. The genetic distances show that populations of the states of Santa Catarina, Sao Paulo, and Rio de Janeiro are genetically closer (0.032 to 0.083) than populations of Bahia (0.364 to 0.853) based on profiles from 10 distinct isoenzyme loci. The Fst was lower (0.077) when the Bahia population was excluded than when it was included (0.300) in the analyses. The inferred number of migrants per generation was 2.99 individuals among populations from the states of Santa Catarina, Sao Paulo, and Rio de Janeiro and 0.58 migrants per generation among all populations. Results suggest that An. cruzii is a complex of species and that the specimens of state of Bahia can be considered as belonging to a species that is distinct from other three closely-related populations studied. PMID- 15543409 TI - Monitoring of the dissemination of Salmonella in the chicken Frankfurt-sausage production line of a sausage factory in the state of Sao Paulo, Brazil. AB - Poultry meat and its derivatives are among the foodstuffs considered by environmental health authorities to present the highest risks to the public. A total of 185 samples were collected in five monthly batches, from different processing stages in a sausage plant that uses mechanically-deboned chicken meat (MDCM), and tested for the presence of Salmonella. Enrichment was carried out in both Kauffman's tetrathionate broth and Rappaport-Vassiliadis broth and isolation on Salmonella-Shigella agar and brilliant-green agar. Live Salmonella bacteria were isolated from six samples of the raw meat and from the emulsion, in batches three, four, and five, but not from any sample in batches one or two. The six isolated strains were all classified as Salmonella Albany, which has not previously been reported in MDCM. Of the two enrichment broths, Rappaport Vassiliadis gave the better results. The pattern of contamination suggests a probable common source, given that a new supplier was used in the third, fourth, and fifth months. It was also shown that the industrial cooking was effective in preventing Salmonella surviving in the final product. PMID- 15543410 TI - Phlebotomine sand flies in Porteirinha, an area of American visceral leishmaniasis transmission in the State of Minas Gerais, Brazil. AB - A study of the phlebotomine sand fly fauna was carried out in an endemic area of American visceral leishmaniasis (AVL) in the municipality of Porteirinha, in the Brazilian state of Minas Gerais. Captures were performed with CDC light traps in 7 districts, 5 days per month, during 2 consecutive years (January 2000 to December 2001). A total of 3240 sand flies were captured and identified. Sixteen species were found, among which 15 belonged to the genus Lutzomyia and one to the genus Brumptomyia. Lutzomyia longipalpis, a proven vector of AVL, was the predominant species (71.85%) throughout the time period. The interference of climatic factors (temperature, humidity, and rainfall) over the populational dynamics of the sand flies was determined. Statistical analysis of the data showed a significant correlation among the number of phlebotomine sand flies collected, rainfall, and humidity, whereas the effect of temperature was negligible, in that particular region. The amount of collected phlebotomine, the number of human cases, and the prevalence of canine AVL in the districts of Porteirinha are discussed. PMID- 15543411 TI - Production of L-asparaginase by filamentous fungi. AB - L-asparaginase production was investigated in the filamentous fungi Aspergillus tamarii and Aspergillus terreus. The fungi were cultivated in medium containing different nitrogen sources. A. terreus showed the highest L-asparaginase (activity) production level (58 U/L) when cultivated in a 2% proline medium. Both fungi presented the lowest level of L-asparaginase production in the presence of glutamine and urea as nitrogen sources. These results suggest that L-asparaginase production by of filamentous fungi is under nitrogen regulation. PMID- 15543412 TI - Histopathological and ultrastructural effects of delta-endotoxins of Bacillus thuringiensis serovar israelensis in the midgut of Simulium pertinax larvae (Diptera, Simuliidae). AB - The bacterium Bacillus thuringiensis (Bt) produces parasporal crystals containing delta-endotoxins responsible for selective insecticidal activity on larvae. Upon ingestion, these crystals are solubilized in the midgut lumen and converted into active toxins that bind to receptors present on the microvilli causing serious damage to the epithelial columnar cells. We investigated the effect of these endotoxins on larvae of the Simulium pertinax, a common black fly in Brazil, using several concentrations during 4 h of the serovar israelensis strain IPS-82 (LFB-FIOCRUZ 584), serotype H-14 type strain of the Institute Pasteur, Paris. Light and electron microscope observations revealed, by time and endotoxin concentration, increasing damages of the larvae midgut epithelium. The most characteristic effects were midgut columnar cell vacuolization, microvilli damages, epithelium cell contents passing into the midgut lumen and finally the cell death. This article is the first report of the histopathological effects of the Bti endotoxins in the midgut of S. pertinax larvae and the data obtained may contribute to a better understanding of the mode of action of this bacterial strain used as bioinsecticide against black fly larvae. PMID- 15543413 TI - Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials. AB - Freshwater snails belonging to the genus Biomphalaria act as intermediate hosts for the parasite trematode Schistosoma mansoni in Africa and in the neotropical region. Identification of such molluscs is carried out based on morphological characters and the presence of cercariae is verified through squeezing snails between two glass slides or by exposing them to artificial light. However, sometimes, the material collected includes molluscs with decomposed bodies or, yet, only empty shells, which precludes their identification and S. mansoni detection. Due to these difficulties, we have developed a methodology in which DNA may be extracted from traces of organic material from inside shells in order to identify molluscs through polymerase chain reaction and restriction fragment length polymorphism and to detect S. mansoni into these snails, by using low stringency polymerase chain reaction. Species-specific profiles obtained from B. glabrata, B. straminea, and B. tenagophila snails and their shells, maintained in laboratory for ten years, showed the same profiles. S. mansoni profiles showed to be present in shell specimens as far as the eighth week after being removed from aquarium. PMID- 15543414 TI - The use of the polymerase chain reaction and restriction fragment length polymorphism technique associated with the classical morphology for characterization of Lymnaea columella, L. viatrix, and L. diaphana (Mollusca: Lymnaeidae). AB - The specific identification of Lymnaeid snails is based on a comparison of morphological characters of the shell, radula, renal and reproductive organs. However, the identification is complicated by dissection process, intra and interspecific similarity and variability of morphological characters. In the present study, polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) techniques targeted to the first and second internal transcribed spacers (ITS1 and ITS2) rDNA and to the mitochondrial 16S ribosomal gene (16S rDNAmt) were used to differentiate the species Lymnaea columella, L. viatrix, and L. diaphana from some localities of Brazil, Argentina, and Uruguay as well as to verify whether the molecular results corroborates the classical morphological method.PCR-RFLP analysis of the ITS1, ITS2, and 16S using 12 restriction enzymes revealed characteristic patterns for L. columella and L. diaphana which were concordant with the classical morphology. On the other hand, for L. viatrix populations a number of 1 to 6 profiles were generated while morphology provided the species pattern results. PMID- 15543415 TI - Detection of Mycobacterium leprae DNA by polymerase chain reaction in the blood and nasal secretion of Brazilian household contacts. AB - DNA samples from blood and nasal swabs of 125 healthy household contacts was submitted to amplification by polymerase chain reaction (PCR) using a Mycobacterium leprae-specific sequence as a target for the detection of subclinical infection with M. leprae. All samples were submitted to hybridization analysis in order to exclude any false positive or negative results. Two positive samples were confirmed from blood out of 119 (1.7%) and two positive samples from nasal secretion out of 120 (1.7%). The analysis of the families with positive individuals showed that 2.5% (n = 3) of the contacts were relatives of multibacilary patients while 0.8% of the cases (n = 1) had a paucibacilary as an index case. All positive contacts were followed up and after one year none of them presented clinical signs of the disease. In spite of the PCR sensitivity to detect the presence of the M. leprae in a subclinical stage, this molecular approach did not seem to be a valuable tool to screen household contacts, since we determined a spurious association of the PCR positivity and further development of leprosy. PMID- 15543416 TI - HygR and PurR plasmid vectors for episomal transfection of Trypanosoma cruzi. AB - This work describes the development and functional testing of two episomes for stable transfection of Trypanosoma cruzi. pHygD contained the 5'- and 3'- flanking regions of the gene encoding the cathepsin B-like protease of T. cruzi as functional trans-splicing and polyadenylation signals for the hygR ORF. Evidence is presented to support extrachromosomal maintenance and organization as tandem repeats in transfected parasites. pPac was derived from pHygD by replacement of the entire hygR ORF with a purR coding region. The ability to modify pHygD and the availability of the complete DNA sequence make these plasmids useful tools for the genetic manipulation of T. cruzi. PMID- 15543417 TI - Study of the antibody response against Mycobacterium tuberculosis antigens in Warao Amerindian children in Venezuela. AB - This study was aimed at investigating alternate methods for serodiagnosis of tuberculosis (TB), which are needed because bacteriologic diagnosis of childhood TB is difficult. A selection of 80 serum and saliva samples were tested from Warao indigenous children under 15 years of age; 34 high TB suspects (28 positive and 6 negative for the tuberculin skin test, TST) and 46 healthy contact children (32 positive and 14 negative for the TST). Several enzyme-linked immunosorbent assay (ELISA) serological tests were developed to test for Mycobacterium tuberculosis-specific antibodies, including serum IgA, IgG, IgE, and secretory IgA (sIgA) in saliva against 3 specific antigens (PPD, HSP60, 38 kDa). Of these, 2 antigens, PPD and 38 kDa, showed significantly higher reactivity. The sensitivity and specificity of these tests for diagnosis remained limited, between 26.5% and 38.2%, and 77.4% and 97%, respectively. Of all the samples studied and combinations realized between all isotypes and antigens combined with 3 isotypes (anti-PPD IgG, IgE, and anti-38kDa sIgA) managed to detect the largest number of patients, showing an improved sensitivity level of 64.7%, although specificity levels dropped to 81.8%. These results were compared with the Omega diagnostics commercial kit results. The commercial kits showed significantly lower reactivity (sensitivity of 20% and 13.33% to Myco G and Complex Plus, respectively) and a specificity of 100%. This study shows that in indigenous populations of Venezuela, where invasive procedures cannot be used to select samples but evaluation with a chest X-ray for radiological studies is available, the combination of 3 specific isotypes may be a useful tool to increase diagnostic accuracy with pulmonary TB in this population, when used together with clinical and epidemiological criteria. PMID- 15543418 TI - Antileishmanial IgG and IgE antibodies recognize predominantly carbohydrate epitopes of glycosylated antigens in visceral leishmaniasis. AB - The specificity of human antileishmanial IgG and IgE antibodies to glycosylated antigens of Leishmania chagasi was evaluated. An ELISA was performed with soluble leishmanial antigen (SLA) and a panel of 95 sera including samples from patients with subclinical infection (SC) and visceral leishmaniasis (VL), subjects cured of visceral leishmaniasis (CVL), and from healthy individuals from endemic areas (HIEA). Antileishmanial IgG were verified for 18 (40%) of 45 SC subjects (mean absorbance of 0.49 +/- 0.17). All nine sera from VL patients had such antibody (0.99 +/- 0.21), while 11 (65%) of 17 CVL individuals were seropositive (0.46 +/- 0.05). Only three (12%) of 24 HIEA controls reacted in IgG-ELISA. Antileishmanial IgE was detected in 26 (58%) of 45 SC patients (0.35 +/- 0.14), and in all VL patients (0.65 +/- 0.29). These antibodies were also detected in 13(76%) of 17 CVL subjects (0.42 +/- 0.14) while all HIEA controls were seronegative. There was no correlation between antileishmanial IgG and IgE antibody absorbances. Mild periodate oxidation at acid pH of SLA carbohydrates drastically diminished its antigenicity in both IgG and IgE-ELISA, affecting mainly the antigens of 125, 102, 94, and 63 kDa as demonstrated by western immunoblotting. PMID- 15543419 TI - Rabies neutralizing antibody detection by indirect immunperoxidase serum neutralization assay performed on chicken embryo related cell line. AB - The aim of this study was to evaluate the indirect immunoperoxidase virus neutralization (IPVN) and mouse neutralization test (MNT) to detect antibodies against rabies virus from vaccinated dogs and cattle. The IPVN was set up for the ability to measure 0.5 International Units/ml (IU) of antibody required by the World Health Organization and the Office International des Epizooties as the minimum response for proof of rabies immunization. IPVN was developed and standardized in chicken embryo related (CER) cell line when 141 dog and 110 cattle sera were applied by serial five-fold dilutions (1:5, 1:25, 1:125) as well as the positive and negative reference controls, all added in four adjacent wells, of 96-well microplates. A 50 microl amount of CVS32 strain dilution containing 50-200 TCID50/ml was mixed to each serum dilution, and after 90 min 50 microl of 3 x 10(5) cells/mlcell suspension added to each well. After five days of incubation, the monolayers were fixed and the IPVN test performed. The correlation coefficient between the MNT and IPVN performed in CER cells was r = 0.9949 for dog sera (n = 100) and r = 0.9307 for cattle sera (n = 99), as well as good specificity (94.7%), sensitivity (87.5%), and agreement (96.6%) were also obtained. IPVN technique can adequately identify vaccinated and unvaccinated animals, even from low-responding vaccinated animals, with the advantage of low cost and faster then MNT standard test. PMID- 15543420 TI - Evaluation of Mesocyclops annulatus (Copepoda: Cyclopoidea) as a control agent of Aedes aegypti (Diptera: Culicidae) in Argentina. AB - We evaluated the potential of Mesocyclops annulatus as a control agent of Aedes aegypti in La Plata city (Argentina). Mosquito larval survivorship due to predation by these copepods was estimated at weekly intervals during the oviposition period of A. aegypti. Mean weekly A. aegypti larval survivorship in cylindrical plastic containers (12 cm height and 11 cm diameter) with copepods was significantly lower than in control containers. Furthermore, weekly larval survival was negatively correlated with M. annulatus adult density, and approximately 23 adult copepods/container would be a threshold density over which the weekly mosquito larval survivorship approached zero. The copepods were able to persist in all containers during approximately 100 days (in three of them until the end of the experiment: 155 days) without the resource represented by A. aegypti larvae. The predation and persistence observed suggest that M. annulatus is a potential control agent to be considered in biological control programs. PMID- 15543421 TI - Larvicidal activity of essential oils from Brazilian plants against Aedes aegypti L. AB - Aedes aegypti L. is the major vector of dengue fever, an endemic disease in Brazil. In an effort to find effective and affordable ways to control this mosquito, the larvicidal activities of essential oils from nine plants widely found in the Northeast of Brazil were analyzed by measurement of their LC50. The essential oils were extracted by steam distillation and their chemical composition determined by GL-chromatography coupled to mass spectroscopy. The essential oils from Cymbopogon citratus and Lippia sidoides, reported in the literature to have larvicidal properties against A. aegypti, were used for activity comparison. The results show that Ocimum americanum and Ocimum gratissimum have LC50 of 67 ppm and 60 ppm respectively, compared to 63 ppm for L. sidoides and 69 ppm for C. citratus. These results suggest a potential utilization of the essential oil of these two Ocimum species for the control of A. aegypti. PMID- 15543422 TI - Dietary supplementation with apple juice concentrate alleviates the compensatory increase in glutathione synthase transcription and activity that accompanies dietary- and genetically-induced oxidative stress. AB - Increased oxidative stress, which can arise from dietary, environmental and/or genetic sources, contributes to the decline in cognitive performance during normal aging and in neurodegenerative conditions such as Alzheimer's disease. Supplementation with fruits and vegetables that are high in antioxidant potential can compensate for dietary and/or genetic deficiencies that promote increased oxidative stress. We have recently demonstrated that apple juice concentrate (AJC) prevents the increase in oxidative damage to brain tissue and decline in cognitive performance observed when transgenic mice lacking apolipoprotein E (ApoE-/-) are maintained on a vitamin-deficient diet and challenged with excess iron (included in the diet as a pro-oxidant). However, the mechanism by which AJC provided neuroprotection was not conclusively determined. Herein, we demonstrate that supplementation with AJC also prevents the compensatory increases in glutathione synthase transcription and activity that otherwise accompany maintenance of ApoE-/- mice on this vitamin-free diet in the presence of iron. Inclusion of the equivalent composition and concentration of sugars of AJC did not prevent these increases. These findings provide further evidence that the antioxidant potential of AJC can compensate for dietary and genetic deficiencies that otherwise promote neurodegeneration. PMID- 15543423 TI - Sociodemographic differences in dietary habits in a population-based sample of elderly subjects: the 3C study. AB - PURPOSE: Diet plays an important role in maintaining quality of life in ageing and can be influenced by sociodemographic factors. We aimed to describe dietary habits in a large population-based sample of elderly people and to explore how they may vary according to sex, age, educational level and lifestyle. METHODS: 9250 community dwellers aged 65 years and over recruited in three French cities were interviewed about their food habits with a brief frequency questionnaire. For each food considered, the subjects were classified as occasional or regular consumers. Quantities of alcoholic beverages consumed daily were also assessed. Dietary habits were compared for men and women separately according each sociodemographic factor. RESULTS: Women were characterized by a more regular consumption of fruit and vegetables and drunk fewer alcoholic beverages. Older subjects ate meat, fish, cereals, raw vegetables and pulses less regularly. The proportion of regular consumers of fish, raw fruit, raw vegetables and cooked fruit or vegetables and the quantity of alcohol consumed increased with educational level. Subjects living alone were less regular consumers of almost all foods. DISCUSSION: The results show that even in a sub-population of elderly people, dietary patterns can vary greatly according to several factors. Identifying such factors is of value for future analytic studies on nutrition and health in the elderly. In view of dietary recommendations advising a high consumption of fruit and vegetables, subjects living alone and people with a low educational level are particularly at risk and should be encouraged to eat more balanced meals. PMID- 15543424 TI - A 6-month follow-up of nutritional status in institutionalized patients with Alzheimer's disease. AB - OBJECTIVE: A follow up of nutritional status in institutionalized patients with Alzheimer's disease. DESIGN: Observational study. SETTING: Specialized unit for patients with Alzheimer's disease in Dijon hospital, France. SUBJECTS: Fourteen women, aged 72-92 years. ASSESSMENTS: On admission of patients to the specialized unit for dementia, body weight, body mass index, arm muscle circumference (AMC) and triceps skin fold (TSF) were measured. Serum concentrations of albumin, prealbumin, homocysteine, orosomucoide, calcium, folates, vitamins B12 and B6 and C-reactive protein were recorded. The same clinical and biological measurements were repeated at day 30, 90 and 180. RESULTS: Both mean weight and mean BMI increased throughout the study with significant differences between day 0 and day 90, and day 180 (p<0.01). The weight gain was associated with a significant increase in AMC and TSF (p<0.01 and p<0.001, respectively). There was no significant difference for values of both mean serum levels of albumin and prealbumin between day 0 and day 180. Whatever the period, serum concentrations of folates and vitamin B12 were in the normal range, while mean levels of vitamin B6 were lower than the normal range; For all these values, there was no significant difference between day 0, 30, 90, 180. Mean levels of C-reactive protein and orosomucoide and lymphocyte counts were in the normal range both at admission and day 180. No significant difference was noted between value of homocysteine at admission and at day 180. CONCLUSION: In this special care unit, the patients with Alzheimer's disease who take part in daily activities and particularly preparation of the meal don't lose weight. It is certain that future studies recording food intake and energy expenditure are necessary to explain the benefits in the nutritional status observed in patients showing dementia when they are institutionalized in a special unit. PMID- 15543425 TI - The epidemiology of recent involuntary weight loss in the United States population. AB - BACKGROUND: Although recent involuntary weight loss (RIWL) has been associated with mortality, no national studies described the prevalence among the general population, characteristics and long-term outcomes of people with RIWL. METHODS: The authors analyzed data from the NHANES II Mortality Study of 5838 individuals 50-74.9 years old who between 1976-1980 underwent a physical examination that included height and weight measurements, biochemical tests and responded to questions about involuntary weight loss within the past six months. Vital status was determined through 1992. Logistic regression was used to examine characteristics associated with RIWL and Cox proportional hazard modeling was used to measure associations between RIWL and mortality. RESULTS: 13.3% of the population reported RIWL with 6.9% reporting > or = 5% RIWL. Obese individuals were at significantly higher risk of RIWL of > or = 5% compared to those with BMI 19-24.9 (OR=1.57. 95% CI: 1.13, 2.18). Other significant risk factors for RIWL included; poor self-reported health, cancer, high white blood cell count, low albumin and low hemoglobin levels, age and current smoking status. RIWL of > or = 5% was significantly associated with mortality (RR=1.24, 95% CI: 1.01, 1.53). CONCLUSION: In summary, RIWL is fairly common among community-dwelling older adults, occurs disproportionately among obese individuals, is associated with characteristics of poor health and independently associated with mortality. These results indicate that RIWL needs to be considered an adverse health indicator even among obese individuals and despite the absence of several clinical indicators of disease. PMID- 15543426 TI - Energy and nutrient intake of elderly hospitalized patients in a steady metabolic status versus catabolic status. AB - Protein undernutrition enhances frailty and aggravates intercurrent diseases generally observed in elderly patients. Undernutrition results from insufficient food intake and catabolic status. Daily nutrient intakes were explored for hospitalized geriatric patients. Nutrient intake (carbohydrates, lipids, proteins, and calcium) was determined in randomly selected geriatric patients (n=49) over five consecutive days by weighting food in the plate before and after meals. For each geriatric patient, catabolic status and risk factors of undernutrition were considered. Results were compared between patients in a steady status or catabolic status. In steady status patients, protein, lipid and carbohydrate intake but not calcium intake, met recommended dietary allowances (total caloric intake:1535 +/- 370 Cal/day ; protein:1+/- 0.4 g/kg/day ; carbohydrates:55 +/- 7.7 % ; lipids: 30 +/- 6.3 % ; calcium:918 +/- 341 mg/day) . Patients in catabolic status (cardiopulmonary deficiency , neurologic disease , inflammatory process) had lower total caloric intake, lower protein intake and dramatically lower calcium intake (total caloric intake : 1375 +/- 500 Cal/day ; protein :0.9 +/- 0.4 g/kg/day ; carbohydrates : 54 +/- 8.3 % ; lipids : 31 +/-6.2 % ; calcium : 866 +/- 379 mg/day). Nutrient intake was lower in elderly patients hospitalized in short stay care units, perhaps due to failure to recognize suitable nutrient requirements. Protein-caloric undernutrition should be diagnosed early during hospitalization in order to allow appropriate dietary supplementation. However the incidence of protein undernutrition among elderly patients as a cause or a consequence of adverse pathophysiological processes remains a cause of debate. PMID- 15543427 TI - Lifetime physical activity and determinants of estimated bone mineral density using calcaneal ultrasound in older South African adults. AB - Increased levels of physical activity (PA) have been linked to higher peak bone mass and increased bone area. The aim of the study was to identify the association between lifetime and current PA with estimated Bone Mineral Density (BMD) in a population of older, socio-economically disadvantaged South Africans of mixed racial ancestry. METHODS: Estimated BMD and T-scores were measured using calcaneal ultrasound (SAHARATM) in 47 men and 105 women, mean age, 65 +/-7 years. Lifetime PA was assessed using a questionnaire comprised of three activity domains (household, occupational, leisure) during five age epochs (14- 21; 22-34; 35-49; 50-64; > 65 years). Current PA was assessed using the Yale Physical Activity Survey. Peak bone strain units for each activity were estimated, based on impact loading. Confounding factors such as BMI, smoking patterns and nutritional status were also quantified. RESULTS: Men and women had similar mean estimated calcaneal BMD, 0.454+/-0.01 and 0.453+/-0.1g/m2, respectively. The proportion of subjects presenting with apparent osteopenia and osteoporosis was similar in men and women (52% vs. 53% and 7% vs. 6%, respectively). Occupational PA between 14 and 21 years, for men (r=0.35; p=0.034) and 22-34 years for women (r=0.24; p=0.033) was significantly correlated to estimated BMD. There were no other significant associations between any of the PA domains and estimated BMD. Leisure time PA only accounted for 0.8% of total energy expenditure for both the men and women. Individuals who were occupationally active between ages 14 and 34 years, were more likely to remain active throughout life. Smokers had lower estimated BMD and T-scores than non-smokers (p<0.01). Reported alcohol and calcium intake was not correlated to any of the ultrasound parameters. CONCLUSION: Estimated BMD was weakly, but significantly correlated to occupational PA during the years of peak-bone mass accretion, which may be protective against accelerated bone loss in later life. From a public health perspective, these data highlight the importance of quantifying lifetime PA in all domains, including occupation. PMID- 15543428 TI - Mini-nutritional assessment score and the risk for undernutrition in free-living older persons. AB - BACKGROUND: Many scales have been proposed for the brief nutritional assessment of older persons, with the goal of increasing undernutrition detection and the detection of nutritional risk. The Mini-Nutritional Assessment (Guigoz et al., 1994) has been increasingly used worldwide, but its efficacy has been assessed in few countries. OBJECTIVES: This study aimed to assess, through complete clinical evaluation, anthropometric measurements and laboratorial tests, the nutritional state of older persons living in the community and compare the results with the score obtained by the application of the Mini Nutritional Assessment. DESIGN: Forty-two persons aged 60 years or older (55% women, mean age 70.9 years) were studied. All volunteers were submitted to a criterious clinical evaluation, anthropometric measures and laboratorial testing (serum albumin, hemoglobin, lymphocite count, iron and unsatured iron binding capacity). RESULTS: Twenty seven volunteers (64%) were considered eutrophic, 12 volunteers were considered obese (28,6%) and three volunteers were considered undernourished. Thirteen volunteers were classified as in risk of undernutrition by the Mini-Nutritional Assessment score, 29 scored within the normal range and no volunteer was considered to be undernourished. The Mini Nutritional Assessment score was significantly associated with age and with the unsatured iron binding capacity. When compared to the final nutritional diagnosis, the questionnaire showed 100% sensibility and 74.3% specificity. CONCLUSION: This study detected a prevalence of undernutrition in the elderly living in the community similar to those described in developed countries. In this population, the Mini Nutritional Assessment showed to be specially efficient for the detection of nutritional risk. PMID- 15543429 TI - The ER function BiP is a master regulator of ER function. AB - The endoplasmic reticulum (ER) is a command center of the cell that is second only to the nucleus in terms of the breadth of its influence on other organelles and activities. It is a major site of protein synthesis, contains the cellular calcium stores that are an essential component of many signaling pathways, and is the proximal site of a signal transduction cascade that responds to cellular stress conditions and serves to maintain homeostasis of the cell. All eucaryotic cells possess an ER, which can comprise nearly 50% of the membranes of a cell. Its functions can be divided into those that occur on the cytosolic side of the membrane (where protein translation and signal transduction cascades occur) and the luminal space (where most other ER functions take place). Our studies during the past several years have revealed that the ER molecular chaperone BiP is a master regulator of ER function. It is responsible for maintaining the permeability barrier of the ER during protein translocation, directing protein folding and assembly, targeting misfolded proteins for retrograde translocation so they can be degraded by the proteasome, contributing to ER calcium stores, and sensing conditions of stress in this organelle, to activate the mammalian unfolded protein response. PMID- 15543430 TI - Measurement of end points in heart failure trials: jousting at windmills? AB - Advances in the treatment of congestive heart failure, a condition of increasing incidence and prevalence, have been made possible by knowledge gained from randomized clinical trials. The selection of end points in these trials has become a pivotal step in the drug and device approval process. In part because of the success of earlier trials, the number of study subjects required in order to realize an important improvement in survival has increased. This has led to the development of alternative combined and composite end points, often including surrogates for mortality. The limitations of these end points, the specific challenges associated with studies of patients with decompensated and diastolic heart failure, and other issues encountered during and after completion of heart failure trials, are discussed. PMID- 15543431 TI - AAV vectors for hemophilia B gene therapy. AB - Adeno-associated viral (AAV) vector is attracting significant interest for use in gene therapy for genetic diseases, because of its unique and advantageous characteristics, compared to other currently available viral vectors. Eight natural serotypes of AAV have been identified, of which AAV serotype 2 is the one best characterized and most widely used in current gene delivery studies. The application of AAV serotype 2 in hemophilia B gene therapy is a promising development in gene therapy for genetic diseases such as hemophilia. Preliminary studies have demonstrated relation and distinction of host, genome sequences, replication, tropism, packaging of recombinant virions and cross-reactivity of neutralizing antibodies among different serotypes of AAV. This review summarizes the progress of studies in AAV serotypes and pertinent applications in hemophilia B gene therapy. The latest progress in gene delivery of coagulant factor IX (for hemophilia B) using AAV serotype vectors is described in detail. PMID- 15543432 TI - The Institute of Medicine report "Unequal Treatment": implications for academic health centers. AB - BACKGROUND: The United States has achieved dramatic improvements in overall health and life expectancy, largely due to initiatives in public health, health promotion and disease prevention. Academic health centers have played a major role in this effort, given their mission of engaging in research, educating health professionals, providing primary and specialty medical services, and caring for the poor and uninsured. However, national data indicate that minority Americans have poorer health outcomes (compared to whites) from preventable and treatable conditions such as cardiovascular disease, diabetes, asthma, cancer and HIV/AIDS. Two factors contribute heavily to these racial and ethnic disparities in health: minorities are subjected to adverse social determinants, and they are disproportionately represented among the uninsured. In the last twenty years, however, the literature has highlighted the fact that racial and ethnic disparities occur not only in health, but also in health care. The Institute of Medicine Report, "Unequal Treatment." The Institute of Medicine (IOM) was asked to determine the extent of racial and ethnic disparities in health care. Their report, entitled "Unequal Treatment," found that racial and ethnic disparities in health care do exist, and that many sources, including health care systems, health care providers, patients and utilization managers, are contributors. Recommendations from "Unequal Treatment": Implications for Academic Health Centers. The IOM Report, "Unequal Treatment," provides a series of recommendations to address racial and ethnic disparities in health care, targeted to a broad audience (the executive summary and full IOM Report can be found at www.nap.edu under the search heading "Unequal Treatment"). Several of the recommendations speak directly to the mission and roles of academic health centers, and have clear and direct implications for patient care, education, and research. These recommendations include collecting and reporting health care access and utilization data by patient=s race/ethnicity, encouraging the use of evidence-based guidelines and quality improvement, supporting the use of language interpretation services in the clinical setting, increasing awareness of racial/ethnic disparities in health care, increasing the proportion of underrepresented minorities in the health care workforce, integrating cross cultural education into the training of all health care professionals, and conducting further research to identify sources of disparities and promising interventions. CONCLUSION: "Unequal Treatment" provides the first detailed, systematic examination of racial/ethnic disparities in health care, and provides a blueprint for how to address them. The report=s recommendations are broad in scope, yet have direct implications for academic health centers. PMID- 15543433 TI - Anticoagulation in pregnant women with prosthetic heart valves. AB - The combination of heart disease and pregnancy can present a formidable challenge to the clinician entrusted to care for both the mother and fetus. Since most data is retrospective, a definitive prognosis for such a patient may be difficult to obtain. Nevertheless, certain cardiac conditions carry greater risks of maternal mortality than do others. However, even for certain preexisting conditions, a tremendous amount of debate persists with respect to risks during pregnancy and optimal peripartum management. One such area of controversy concerns anticoagulation in pregnant women with prosthetic heart valves. For patients who require anticoagulation for mechanical valves, the choice of some combination of warfarin, unfractionated heparin, and low-molecular-weight heparin (LMWH) has resulted in many small-scale trials, which have not yet provided definite guidance as to the best course of action. Even more controversial has been the recent labeling change that advises against the use of LMWH in all patients with prosthetic heart valves, as a result of two cases of prosthetic valve thrombosis in women using LMWH while pregnant. Although the latest product labeling, in the summer of 2003, was changed to a less restrictive recommendation, debate persists. A discussion of the available data on anticoagulation in pregnant women with prosthetic heart valves is presented here, to inform the clinician and the patient of the risks and benefits of the options presently available. PMID- 15543434 TI - Acute myocardial infarction and left subclavian artery occlusion in Behcet's disease: a case report. AB - A 42-year-old woman with Behcet's disease and pericardial effusion for 14 years presented with acute myocardial infarction and received thrombolytic therapy. Coronary angiography showed total occlusion of the left anterior descending artery. Subsequently, the left internal mammary artery was grafted to the left anterior descending artery. Approximately one year after bypass surgery, digital subtraction angiography of the left subclavian artery, performed because of chest and left arm pain, showed total occlusion of the left subclavian artery. Retrograde flow from the left vertebral artery filled the distal portion of the left subclavian artery. In conclusion, patients with Behcet's disease should be investigated closely for involvement of other arteries when one vessel's involvement has been detected. We also recommend that free arterial grafts be used for any coronary surgical intervention, because of the risk of occlusion associated with the vasculitis of Behcet's disease. PMID- 15543435 TI - Physicians' religiosity and end-of-life care attitudes and behaviors. AB - BACKGROUND: Physicians play the central role in decisions to initiate, withhold and withdraw life-sustaining medical care. Prior studies show that physicians= religiosity is related to end-of-life care attitudes and practices, which if not in concert with the patient or family may be a source of conflict. We surveyed physicians of one religion to describe the relationship between religiosity and end-of-life care. METHODS: Cross-sectional survey of 443 Jewish physicians at four Israeli hospitals, which characterized religiosity and asked about attitudes and communication with patients about end-of-life issues and care practices. RESULTS: Very religious physicians, compared to moderately religious and secular physicians, were much less likely to believe that life-sustaining treatment should be withdrawn (11% vs. 36% v. 51%, p<0.001), to approve of prescribing needed pain medication if it will hasten death (69% vs. 80% vs. 85%, p<0.01), or to agree with euthanasia (5% vs. 42% vs. 70%, p<0.001). Religiosity was not related to withholding most life-sustaining treatments, but even after adjustment for physician and practice characteristics, very religious physicians were much less likely to "ever stop life-sustaining treatment provided to a suffering terminally ill patient" (p<0.0003). Religiosity was unrelated to physician patient communication or to desire for support concerning end-of-life care. Desire for support was universally high. CONCLUSIONS: Physicians' religiosity can have a major effect on the way their patients die, including whether patients receive adequate analgesia near death. Patients may need to query physicians' religious perspectives to ensure that they are consistent with patients' end-of life care preferences. Evaluation of religiosity-related clinical behavior in other cultures is needed. PMID- 15543436 TI - Fibrous pleural tumor with hypoglycemia: case study. AB - Many neoplastic tumors exhibit paraneoplastic syndromes manifested by endocrinopathy. This is particularly true of intrathoracic tumors such as lung cancers, thymomas, carcinoid tumors and mediastinal germ cell neoplasm. Fibrous tumors of the pleura are rare intrathoracic tumors, which are usually benign and often grow to huge size. A subset of these neoplasms present with the syndrome of hypoglycemia. Although first reported more than 70 years ago, the diagnosis is rarely considered when a patient presents with syncope and hypoglycemia. This article reports a patient who presented with a large pleural mass and a hypoglycemic syndrome. (The disease was surgically cured.) The probable mechanism of hypoglycemia is discussed. PMID- 15543437 TI - Cytomegalovirus colitis in an immunocompetent patient with amebiasis: case report and review of the literature. AB - Most cases of cytomegalovirus colitis occur in adults with severe immune deficiency. Only a few cases involving immunocompetent patients have been reported. We describe the first case reported in English, of cytomegalovirus colitis in an immunocompetent patient with preceding amebiasis. Eight previous cases have been reported of cytomegalovirus colitis occurring after colonic mucosal injury in immunocompetent patients. Similar to our case, all eight of these cases resolved without the use of antiviral therapy. This suggests that disruption of colonic mucosa may predispose immunocompetent patients to cytomegalovirus colitis, and that the colitis may become self-limited once the preceding intestinal damage resolves. PMID- 15543438 TI - Liver abscess in Crohn's disease: a report of three cases. AB - Liver abscess is a rare complication of Crohn's disease and must be considered part of the differential diagnosis. We present three case reports and a review of the literature. The incidence of liver abscess is higher in patients with Crohn's disease than in the general population. Since Crohn's disease patients who present with a liver abscess generally do not respond to medical management alone, surgical or percutaneous drainage of the abscess is necessary. PMID- 15543439 TI - "Thou shalt not kill": some legal and linguistic problems. AB - Large plaques of the religious precepts generally known as the Ten Commandments were recently placed in official public spaces in several states. This practice has been successfully challenged in federal courts of appeals, and the challenge was upheld by the Supreme Court in April 2003. Yet there is another problem with such plaques, if older translations are used. "Thou shalt not kill" is actually a mistranslation of the Hebrew, "You shall not murder. The two statements are not synonymous, yet many people in the world still use archaic biblical translations, with unfortunate bioethical consequences. There is also widespread usage of a non commandment, "Thou shalt not kill; but needst not strive officiously to keep alive." This is not a biblical injunction; it is a line from A.H. Clough's satirical poem, "The Latest Decalogue." PMID- 15543440 TI - Eighty hours a week: a third-year resident's perspective on duty-hour limitations. PMID- 15543441 TI - "Stroke, stroke": a coxswain's call for more work and more innovation. PMID- 15543442 TI - Automated Constraint-Induced Therapy Extension (AutoCITE) for movement deficits after stroke. AB - We report progress in the development of AutoCITE, a workstation that delivers the task practice component of upper-limb Constraint-Induced Movement therapy and that can potentially be used in the clinic or the home without the need for one on-one supervision from a therapist. AutoCITE incorporates a computer and eight task devices arranged on a modified cabinet. Task performance is automatically recorded, and several types of feedback are provided. In preliminary testing, nine chronic stroke subjects with mild to moderate motor deficits practiced with AutoCITE for 3 h each weekday for 2 weeks. Subjects wore a padded mitt on the less-affected hand for a target of 90% of their waking hours. In terms of effect sizes, gains were large and significant on the Motor Activity Log, and moderate to large on the Wolf Motor Function Test. These gains were comparable to the gains of a matched group of 12 subjects who received standard Constraint-Induced Movement therapy. PMID- 15543443 TI - The experience of time in the transition from hospital to home following stroke. AB - This paper reports findings related to the transition from hospital to home during the first month after discharge following acute stroke. Qualitative data were obtained from in-depth, semistructured interviews with 51 male stroke survivors and their caregivers. Data were analyzed with the N6 software application, designed to assist with qualitative data analysis. Stroke survivors described experiences related to changes in the temporal order of life, disruptions in sense of self, and strategies used to manage time. Findings indicate that changes in the temporal order of life are related to functional impairments and disruption in the taken-for-granted body. At 1 month post discharge, survivors are struggling with establishing routines in their day and coping with an increased amount of idle time. In conclusion, strategies for managing increased idle time are "passing time," "waiting on time," and "killing time." PMID- 15543444 TI - Why ethnic designation matters for stroke rehabilitation: comparing VA administrative data and clinical records. AB - Using existing administrative data to look at issues of ethnic disparities in rehabilitation-related outcomes may lead to misleading results. Problems can emerge from apparently small issues of reliability that are magnified by reclassification of ethnic designation and missing data in complete-subject analyses. We compared the reliability of ethnic assignment in Department of Veterans Affairs (VA) medical rehabilitation records for stroke patients with administrative records; reclassified the racial identifier from the administrative data in two ways; and examined the different sources of ethnic information in relation to severity, length-of-stay, disability assessment, and discharge disposition. Our results show how small changes increase the potential for Type II error when describing ethnic differences in outcomes or using ethnicity as a predictor with dichotomous response variables. We discuss our results with reference to the literature on ethnic classification and underline the importance of initiatives for improved data collection on ethnicity in VA data sources and in rehabilitation research. PMID- 15543445 TI - How strong is the relationship between functional status and quality of life among persons with stroke? AB - The quantitative relationship between functional status and self-reported quality of life is relatively unexamined. As part of the 1-, 6-, and 12-month telephone follow-up of consecutive patients in an observational study of patients with stroke, we found that while higher functional status was associated with better quality of life, this relationship was relatively weak (Spearman correlation <0.25). Patients with similar levels of disability reported quite different qualities of life. Any improvement in quality of life over time was modest at best. Mean utilities for patients with minor stroke were near 0.80, while those for patients with major stroke were near 0.60, the latter figure exceeding previous reports. Quality of life with major stroke may not necessarily be as low as that reported before such a stroke occurs. Quality of life after stroke is heterogeneous and depends on more than just level of physical function. PMID- 15543446 TI - Stepping over obstacles to improve walking in individuals with poststroke hemiplegia. AB - For this study, we evaluated two training interventions for improving gait parameters in individuals with poststroke hemiplegia using a training methodology that required them to step over objects. Gait velocity, step length, ability to step over obstacles, and walking endurance were compared before and after 2 weeks of training and 2 weeks after cessation of training. Twenty subjects with poststroke hemiplegia completed six intervention sessions in which they were asked to step over either virtual objects while walking on a motorized treadmill or real foam objects on a 10 m walkway. With the virtual object training, if either foot collided with the virtual object, a tone sounded and a vibrotactile stimulus was applied to the colliding foot. All subjects tolerated the training sessions well, and no incidences of falling or undue cardiovascular responses occurred. The virtual obstacle training generated greater improvements in gait velocity compared with real training (20.5% vs. 12.2% improvement) during the fast walk test (p < 0.01). Improvements in gait velocity for both training methods were similar in the self-selected walk test (33.3% vs. 34.7% improvement). Overall, subjects showed clinically meaningful changes in gait velocity, stride length, walking endurance, and obstacle clearance capacity as a result of either training method. These changes persisted for 2 weeks posttraining. The inclusion of enhanced safety and visual augmentation may be responsible for the effectiveness of the virtual object intervention. These results demonstrate preliminary evidence for clinical effectiveness of obstacle training for improving gait velocity poststroke. In addition, these results provide evidence for enhanced clinical performance with virtual obstacle training. PMID- 15543447 TI - Weakness and strength training in persons with poststroke hemiplegia: rationale, method, and efficacy. AB - Several converging lines of contemporary evidence suggest that weakness presents a more serious compromise to movement function in poststroke hemiplegia than spasticity. This review examines the clinical and functional phenomena of weakness in poststroke hemiplegia, currently available evidence identifying physiologic substrates contributing to weakness, and reports of early investigations involving high-resistance training targeted at improving strength and the transfer of strength to improvements in functional capacity. Based on this information, we describe some unsolved problems and indicate some likely lines of development to increase our knowledge regarding how resistance training can be included in effective stroke rehabilitation. PMID- 15543448 TI - Application of adaptive filters to visual testing and treatment in acquired pendular nystagmus. AB - Acquired pendular nystagmus (APN) complicates multiple sclerosis and other neurological disorders, causes visual impairment, and frequently resists treatment. Vision could be improved by a visual aid that gates or shifts the seen world in lockstep with the APN. Since the pathological oscillations are embedded in normal eye movements, such a device must track the nystagmus selectively. We evaluated the ability of an adaptive filter to perform this tracking and improve acuity when coupled to either of two devices--a shutter that permitted brief glimpses of the world synchronized with the nystagmus, or simulated image shifting optics. In 10 normal subjects whose decimal acuity averaged 1.46 +/- 0.20, acuity fell to 0.36 +/- 0.08 under viewing conditions simulating APN. The synchronized shutter restored acuity to 0.60 +/- 0.12, while image-stabilization raised it to 1.17 +/- 0.13. Adaptive filters provide a practical means by which to track nystagmus. The most effective visual aid would couple such filters to image-stabilizing optics. PMID- 15543449 TI - Image-shifting optics for a nystagmus treatment device. AB - Acquired pendular nystagmus (APN) complicates multiple sclerosis and other neurological disorders, causes visual impairment, and is often refractory to available treatments. Vision could be improved by an optical aid that shifts the seen world in lockstep with the APN. An essential component of such a device is the image-shifting mechanism, which must be light, accurate, suitable for battery operation, and capable of image shifting at the frequencies and amplitudes seen in APN. We determined that a three-lens image-shifting mechanism used in commercial image-stabilizing lenses has the potential to satisfy all these requirements. In combination with software designed to track nystagmus, the optical mechanism proved capable of improving visual acuity in 12 normal subjects experiencing simulated two-dimensional nystagmus. Acuity was restored to within an average of 0.12 logMAR (range 0.0-0.22) of the subjects' values without the simulated nystagmus. These results support the feasibility of an assistive device for patients with APN. PMID- 15543450 TI - Mobility function in older veterans improves after blind rehabilitation. AB - This study was conducted to investigate the effects of blind rehabilitation training on self-reported mobility function in visually impaired adults. Mobility function was assessed with a questionnaire administered before and 2 months after subjects completed a comprehensive blind rehabilitation program that included orientation and mobility training. Subjects rated the level of difficulty performing in 26 of 34 mobility situations as significantly lower after rehabilitation. Subjects also rated their confidence as higher after rehabilitation. Substantial improvement occurred in the self-reported mobility function of visually impaired adults after blind rehabilitation and mobility training. PMID- 15543451 TI - Relationship of retinal structural and clinical vision parameters to driving performance of diabetic retinopathy patients. AB - This study identifies clinical vision measures or retinal structural measures associated with the driving performance of diabetic retinopathy patients. Twenty five licensed drivers with diabetic retinopathy (median age, 53 years; range, 34 72 years) completed clinical tests (visual acuity, letter contrast sensitivity, and Humphrey 30-2 visual fields) and structural examinations (retinal thickness analysis and fundus photograph grading of retinopathy and laser scarring). Driving performance was assessed with an interactive driving simulator and a driving history questionnaire. Increased retinal thickness was significantly correlated with a higher frequency of simulator accidents and near accidents. Laser scar grades significantly correlated with steeper brake-response slopes, increased brake-pressure standard deviation (SD), and longer response times. Subjects with focal laser scars had significantly higher average brake-pedal pressure and brake-pressure SD than subjects without focal laser scars. Retinal thickness and laser scarring correlated with driving simulator performance in subjects with diabetic retinopathy. PMID- 15543452 TI - Multidimensional visual field maps: relationships among local psychophysical and local electrophysiological measures. AB - Multidimensional psychophysical and electrophysical maps of the central retina are essential for assessing the functioning of the diseased retina. In this study, grating acuity, contrast sensitivity, duration for letter identification, multifocal electroretinograms, and Humphrey visual field thresholds were measured at equivalent positions throughout the central 20 degrees. We found that the rates of sensitivity loss were not equivalent for all psychophysical measures. The rate of loss in the duration required for letter identification as a function of eccentricity was the steepest, followed by acuity and contrast sensitivity. The rate of loss in luminance sensitivity as measured in the Humphrey visual field was the shallowest. The pattern of losses also varied across meridians. Specifically, the rate of loss as a function of eccentricity was highest in the vertical meridian and lowest in the horizontal meridian. These maps and the correlations among measures as a function of retinal position serve as a baseline so that we can examine disease effects throughout the retina. In addition, the development of vision rehabilitation programs focused on eccentric viewing training should consider the differential sensitivities of the peripheral retina. PMID- 15543453 TI - Use of auditory brainstem responses for the early detection of ototoxicity from aminoglycosides or chemotherapeutic drugs. AB - Effective objective HF (high-frequency) testing methodology provides for the early detection of ototoxic hearing loss because it typically progresses from high to low frequencies. Such early detection is considered necessary to prevent hearing loss from progressing into the frequency range important for understanding speech. Objective tests must be reliable, sensitive to hearing change, and time efficient. Auditory brainstem responses (ABRs) appear well suited to this task; however, current ABR techniques have limitations. Conventional clicks stimulate middle (1-4 kHz) rather than high frequencies (>8 kHz). Responses to HF tone bursts require considerable recording time. We hypothesized that using HF band-limited clicks (HF clicks) could overcome these limitations. Two different HF clicks, with bandwidths of 8-14 kHz were used to elicit ABRs. The current study compared responses among these stimuli. The results demonstrate the reliability of HF-click responses and of tone bursts presented in trains. PMID- 15543454 TI - Optimizing care for combat amputees: Experiences at Walter Reed Army Medical Center. PMID- 15543455 TI - A report on the amputee healthcare and prosthetics workshop sponsored by Walter Reed Army Medical Center and the Department of Veterans Affairs (VA) November 17 18, 2003. PMID- 15543456 TI - The biomechanics of wheelchair propulsion in individuals with and without upper limb impairment. AB - We used an instrumented wheelchair ergometer and 3D motion analysis system to collect joint kinematic and temporal data, as well as hand rim and joint kinetics, in 47 manual wheelchair users (MWCUs) (15 with upper-limb impairment and 32 without upper-limb impairment). The group with upper-limb impairment propelled with a higher stroke frequency and reduced hand-rim contact time, and smaller peak joint angles and joint excursion of the wrist, elbow, and shoulder during the contact phase. They also propelled with a reduced power output and reduced hand-rim propulsive and resultant forces, moments, and joint compressive forces. We concluded that these kinematic and kinetic strategies might be a mechanism for allowing MWCUs with upper-limb impairment to remain independent. Additionally, the reduced joint excursion and reduced magnitude of forces may protect them from the development of secondary upper-limb pathologies. PMID- 15543457 TI - Prevalence and identification of shoulder pathology in athletic and nonathletic wheelchair users with shoulder pain: A pilot study. AB - Although many wheelchair users report shoulder pain, the prevalence of specific pathologies remains controversial. Rotator cuff impingement, glenohumeral instability, and biceps tendonitis have been stated as the most commonly found pathology. This study investigated the prevalence and identity of shoulder pathology in athletic and nonathletic manual wheelchair users (MWCUs). Fifty-two MWCUs (26 athletes, 26 nonathletes) completed a survey regarding the nature of their injury, sports involvement, history, and presence of current and/or past shoulder pathology. Subjects currently experiencing shoulder pain underwent a clinical examination of both shoulders. Analysis of variance (p > GGPP > FPP. The DOC activated WBM synthesized less polyprenyl intermediates (butanol extractable) but more final rubber product (toluene/hexane extract), different than FPP and GGPP. WBM enzymes were highly versatile in using diverse different allylics, but UPP was most preferable. WRP was found a little active for UPP with DOC, but still much lower than WBM. Rubber product analysis by RP-TLC with acetone/hexane solvent system showed that WBM was mostly rubber, but WRP was mainly the intermediates. Quantitative analysis showed that WBM labeled rubber was confined to the origin spot, different than WRP as mainly labeled intermediates. It was thus confirmed that the WBM plays the key role in RB functions, and not WRP as mostly reported. WBM served as the actual rubber synthesis site, and bacterial UPP was very good RB initiator. PMID- 15543543 TI - Physical characterization of coupled poly(lactic acid)/starch/maleic anhydride blends plasticized by acetyl triethyl citrate. AB - Acetyl triethyl citrate (ATC) was used as a plasticizer for poly(lactic acid) (PLA)/starch blends coupled with maleic anhydride and an initiator of 2,5 bis(tert-butylperoxy)-2,5-dimethylhexane (L101). Elongation of the blend at break was markedly increased when the ATC content was above the 8% loading level, which is referred to as the percolation threshold. The extended elongation was achieved at the expense of tensile strength and elastic modulus. Thermal transitions of the blend, including the glass transition temperature (T(g)), cold crystallization temperature (T(c)) and melting temperature (T(m)), decreased with ATC content. Thermally induced ATC migration affected the thermal behavior of the plasticized blends and reduced elongation and tensile strength, whereas the elasticity modulus increased. ATC migration increased with ambient temperature, which was controlled by the activation energy of the blend system. Leaching of ATC was slow at room temperature in distilled water, but significant in boiling water. Additionally, the leaching rate was also directly proportional to the ATC content of the blend. Glass transition temperatures of PLA/starch/MA/L101 blends with various acetryl triethyl citrate contents. PMID- 15543544 TI - False diagnosis of renal agenesis on fetal MRI. AB - We report a twin pregnancy in which prenatal ultrasound revealed anhydramnios and bilateral absence of the kidneys in both fetuses. To overcome the difficulties faced in obtaining accurate information on fetal ultrasound in cases with oligohydramnios, an attempt was made to use fetal magnetic resonance imaging (MRI) to confirm the renal status. However, while the fetal MRI showed renal agenesis in both twins, postnatal follow-up revealed that one twin, which survived, had a small unilateral kidney not seen antenatally on fetal ultrasound or MRI. The second twin had bilateral renal agenesis and died of pulmonary hypoplasia. Care should be taken when using fetal MRI to replace fetal ultrasound in cases with poor visibility. PMID- 15543545 TI - Anogenital malformation with ambiguous genitalia as part of the OEIS complex. PMID- 15543546 TI - Mobile, outdoor continuous-flow isotope-ratio mass spectrometer system for automated high-frequency 13C- and 18O-CO2 analysis for Keeling plot applications. AB - A continuous-flow isotope-ratio mass spectrometer (CF-IRMS, custom-made GasBenchII and Delta(plus)Advantage, ThermoFinnigan) was installed on a grassland site and interfaced with a closed-path infrared gas analyser (IRGA). The CF-IRMS and IRGA were housed in an air-conditioned travel van. Air was sampled at 1.5 m above the 0.07-m tall grassland canopy, drawn through a 17-m long PTFE tube at a rate of 0.25 L s(-1), and fed to the IRGA and CF-IRMS in series. The IRMS was interfaced with the IRGA via a stainless steel capillary inserted 0.5 m into the sample air outlet tube of the IRGA (forming an open split), a gas-tight pump, and a sample loop attached to the eight-port Valco valve of the continuous-flow interface. Air was pumped through the 0.25-mL sample loop at 10 mL s(-1) (a flushing frequency of 40 Hz). Air samples were analysed at intervals of approx. 2.8 min. Whole system precision was tested in the field using air mixed from pure CO2 and CO2-free air by means of mass flow controllers. The standard deviation of repeated single measurements was 0.21-0.07 per thousand for delta13C and 0.34 0.14 per thousand for delta18O of CO2 in air with mixing ratios ranging between 200-800 micromol mol(-1). The CO2 peak area measured by the IRMS was proportional to the CO2 mixing ratio (r2 = 1.00), allowing estimation of sample air CO2 mixing ratio from IRMS data. A 1-day long measurement cycle of CO2, delta13C and delta18O of air sampled above the grassland canopy was used to test the system for Keeling plot applications. Delta18O exhibited a clear diurnal cycle (4 per thousand range), but short-term (1-h interval) variability was small (average SD 0.38 per thousand). Yet, the correlation between delta18O and CO2 mixing ratio was relatively weak, and this was true for both the whole data set and 1-h subsets. Conversely, the delta13C of all 541 samples measured during the 25.2-h interval fitted well the Keeling regression (r2 = 0.99), yielding an intercept of -27.40 per thousand (+/-0.07 per thousand SE). Useful Keeling regressions (r2 > 0.9, average r2 = 0.96) also resulted from data collected over 1-h intervals of the 12-h long twilight and dark period. These indicated that 13C content of ecosystem respiration was approx. constant near -27.6 per thousand. The precision of the present system is similar to that of current techniques used in ecosystem studies which employ flask sampling and a laboratory-based CF-IRMS. Sampling (and measurement) frequency is greatly increased relative to systems based on flask sampling, and sampling time (0.025 s per sample) is decreased. These features increase the probability for sampling the entire CO2 range which occurs in a given time window. The system obviates sample storage problems, greatly minimises handling needs, and allows extended campaigns of high frequency sampling and analysis with minimal attendance. PMID- 15543547 TI - Selective determination of organophosphate flame retardants and plasticizers in indoor air by gas chromatography, positive-ion chemical ionization and collision induced dissociation mass spectrometry. AB - Gas chromatography/ion trap mass spectrometry with in-source ionization and dissociation was used in positive-ion chemical ionization (PICI) mode for the determination of organophosphate triesters in indoor air. These compounds are widely used as additive flame retardants and plasticizers in different types of materials and have become ubiquitous pollutants in indoor environments. When using collision-induced dissociation in PICI mode the fragmentation of the organophosphate triesters can be performed in a more controllable way than in electron ionization (EI) mode. The developed selected-reaction monitoring method provided high selectivity for the investigated compounds. For 8-h air measurements (corresponding to 1.5 m3 of sampled air) the limit of detection of the method was determined to be in the range 0.1-1.4 ng m(-3), which is comparable with nitrogen-phosphorus detection and about 50-fold lower than when using EI in selected-ion monitoring mode. The presented method was applied to samples from three common indoor environments, in which a number of organophosphate triesters were identified and quantified. The dominating compound was found to be tris(2-chloropropyl) phosphate, which occurred at levels up to 0.8 microg m(-3). PMID- 15543548 TI - Estimation of the intervention effect in a non-randomized study with pre- and post-mismeasured binary responses. AB - In non-randomized clinical studies, the regression phenomenon can confound interpretation of the effectiveness of an intervention. The regression effect arises due to daily variation and/or misclassification of the biologic marker used in selection as well as in the assessment of the intervention effect. We consider a scenario in which the selection criterion for a subject's participation in the study is such that he/she must have a positive diagnostic test at screening. The disease status is then reassessed at the end of intervention. Thus, two repeated measurements of a binary disease outcome are available, with only selected subjects having a second measurement upon follow up. We propose methods for estimating the change in event probability resulting from implementing the intervention while adjusting for the misclassification that produces the regression effect. We extend this approach to estimation of both the placebo and intervention effects in placebo-controlled studies designed with a misclassified binary outcome. Analyses of two biomedical studies are used for illustration. PMID- 15543549 TI - Mass spectral study of alkali-cationized Boc-carbo-beta3-peptides by electrospray tandem mass spectrometry. AB - Electrospray tandem mass spectrometry was used to study the dissociation reactions of [M+Cat]+ (Cat = Na+ and Li+) of Boc-carbo-beta3-peptides. The collision-induced dissociation (CID) spectra of [M+Cat-Boc]+ of these peptides are found to be significantly different from those of [M+H-Boc]+ ions. The spectra are more informative and display both C- and N-terminus metallated ions in addition to characteristic fragment ions of the carbohydrate moiety. Based on the fragmentations observed in the CID spectra of the [M+Cat-Boc]+ ions, it is suggested that the dissociation involves complexes in which the metal ion is coordinated in a multidentate arrangement involving the carbonyl oxygen atoms. The CID spectra of [M+Cat-Boc]+ ions of the peptide acids show an abundant N terminal rearrangement ion [b(n)+17+Cat]+ which is absent for esters. Further, two pairs of positionally isomeric Boc-carbo-beta3-peptide acids, Boc-NH-Caa(S) beta-hGly-OH (11) and Boc-NH-beta-hGly-Caa(S)-OH (12), and [Boc-NH-Caa(S)-beta hGly-Caa(S)-beta-hGly-OH] (13) and [Boc-NH-beta-hGly-Caa(S)-beta-hGly-Caa(S)-OH] (14), were differentiated by the CID of [M+Cat-Boc]+ ions. The CID spectra of compounds 11 and 13 are significantly different from those of 12 and 14, respectively. The abundance of [b(n)+17+Cat]+ ions is higher for peptide acids 12 and 14 with a sugar group at the C-terminus when compared to 11 and 13 which contain a sugar moiety at the N-terminus. The observed differences between the CID spectra of these isomeric peptides are attributed to the difference in the preferential site of metal ion binding and also on the structure of the cyclic intermediate involved in the formation of the rearrangement ion. PMID- 15543550 TI - Lymphoma and peripheral neuropathy: a clinical review. AB - Lymphoma occasionally affects the peripheral nervous system. When it does, the diagnosis can be elusive since many patients present without known lymphoma. Most peripheral nerve complications are due to non-Hodgkin's lymphoma (NHL), which infiltrates nerves causing axonal damage. This disorder can affect nerve roots and cranial nerves, often associated with lymphomatous meningitis. NHL may also infiltrate peripheral nerves and cause plexopathy, mononeuropathy, or generalized neuropathy. These neuropathies may resemble an asymmetric mononeuropathy multiplex or a generalized disorder such as chronic inflammatory demyelinating polyradiculoneuropathy. When NHL infiltrates diffusely, the term neurolymphomatosis is used. Hodgkin's lymphoma (HL), by contrast, rarely infiltrates nerves. More often, HL causes immunological disorders of the peripheral nervous system such as inflammatory plexopathy or Guillain-Barre syndrome. Other rare lymphomas such as intravascular lymphoma and Waldenstrom's macroglobulinemia can also affect peripheral nerves in specific ways. In addition, other malignant and nonmalignant lymphoproliferative disorders enter into the differential diagnosis of lymphomatous neuropathy. This review discusses the multiple peripheral nerve presentations of lymphoma from the clinician's point of view and provides a guide to the evaluation and diagnosis of these uncommon, challenging disorders. PMID- 15543551 TI - Electrophysiological signs of permanent axonal loss in a follow-up study of patients with Guillain-Barre syndrome. AB - The neurophysiological mechanisms for persisting impairment of motor function after Guillain-Barre syndrome (GBS) were assessed in 37 unselected patients 1-13 years after diagnosis. For evaluation of reinnervation and axonal loss, macroelectromyography (macro-EMG) including measurement of fiber density (FD) was performed. Data from neuropathy symptom score, neuropathy disability score, nerve conduction studies, and quantitative sensory examination were ranked and summed to a neuropathy rank sum score (NRSS). The isokinetic muscle strength at the ankle was measured. Signs of axonal loss with increase of either macro motor unit potential (macro-MUP) amplitude or FD occurred in 76% of patients. The macro-MUP amplitude correlated with muscle strength and with NRSS. Patients with evidence of residual neuropathy had increased macro-MUP amplitude and FD as well as decreased muscle strength compared to patients without evidence of residual neuropathy. We conclude that axonal loss takes place in a substantial number of GBS patients and is associated with permanent muscle weakness caused by insufficient reinnervation. Possible patterns of pathology are discussed in relation to the macro-EMG findings. PMID- 15543552 TI - Activity patterns of wrist extensor muscles during wrist extensions and deviations. AB - Wrist extensor muscles are prone to certain focal musculoskeletal disorders for which the activation pattern of the extensor carpi radialis (ECR) and ulnaris (ECU) muscles may be important risk factors. Surface and intramuscular EMG of these muscles were recorded during isometric low-force wrist extension in semipronation and pronation as well as for ulnar/radial deviation, and were analyzed using root mean square (RMS) and decomposition methods. Despite shorter ECR length at semipronation, higher amplitudes of intramuscular EMG and of motor unit action potentials (MUAPs) were found in pronation than in semipronation. However, these changes were not detectable in the surface EMG. Higher ECR activity levels were also found during wrist extension compared to ulnar/radial deviation, and differences in the motor unit (MU) properties were found during ulnar deviation compared to radial deviation and extension. Remarkably, the MUAPs of ECR were almost twice as large as those of the ECU. Overall, the ECR muscle did not respond as predicted from biomechanical considerations, and in general activity level was higher than expected. This may partly explain why the tendon of the ECR often is associated with lateral epicondylitis. PMID- 15543553 TI - Magnetic resonance study of the influence of tissue damage and cortical reorganization on PASAT performance at the earliest stage of multiple sclerosis. AB - We sought to determine the influence of tissue damage and the potential impact of cortical reorganization on the performance to the Paced Auditory Serial Addition Test (PASAT) in patients at the earliest stage of multiple sclerosis (MS). Magnetization transfer ratio (MTR) imaging and functional magnetic resonance imaging (fMRI) experiments using PASAT as paradigm were carried out in 18 patients with clinically isolated syndrome suggestive of MS (CISSMS) compared to 18 controls. MTR histogram analyses showed structural abnormalities in patients involving the normal-appearing white matter (NAWM) but also the gray matter (GM). Mean PASAT scores were significantly lower in the group of patients taken as a whole, and were correlated with the mean NAWM MTR value. No correlation was observed between PASAT scores and GM MTR. However, in the subgroup of patients with normal PASAT performance (n = 9), fMRI showed larger activations in bilateral Brodmann area 45 (BA45) and right BA44 compared to that in controls (n = 18). In these areas with potentially compensatory reorganization, the whole group of patients (n = 18) showed significantly greater activation than controls (n = 18). Activation in the right BA45 was inversely correlated with the mean NAWM MTR and the peak position of GM MTR histograms of patients. This study indicates that even at the earliest stage of MS, cortical reorganization is present inside the executive system of working memory and could tend to limit the determinant functional impact of NAWM injury on the execution of the PASAT. PMID- 15543554 TI - Distinct neural systems underlie learning visuomotor and spatial representations of motor skills. AB - Motor skill learning depends upon acquiring knowledge about multiple features of sequential behaviors, including their visuomotor and spatial properties. To investigate the neural systems that distinguish these representations, we carried out functional magnetic resonance imaging (fMRI) as healthy adults learned to type sequences on a novel keyboard. On the initial training day, learning-related changes in brain activation were found in distributed cortical regions, only a subset of which correlated with improvements in movement time (MT), suggesting their preeminence in controlling movements online. Subjects received extended training on the sequences during the ensuing week, after which they returned to the scanner for another imaging session. Relative to performance at the end of the first training day, continued plasticity was most striking in the inferior parietal cortex and new areas of plasticity were uncovered in the caudate and cerebellum. Plasticity in these regions correlated with reaction time (RT), suggesting their role in planning sequences before movement onset. Two transfer conditions probed for "what" subjects learned. The probe for visuomotor learning produced increased activation in visual analysis (left inferior visual cortex) and advance planning (left caudate) systems. The probe for spatial learning produced increased activation in visuomotor-transformation (left dorsal visual pathway) and retrieval (left precuneus) systems. Increased activity in all of these regions correlated with increased RT, but not MT, indicating that both transfer conditions interfered with the neural representation of plans for the sequences, but not processes that controlled their implementation. These findings demonstrated that neuroanatomically dissociable systems support the acquisition of visuomotor and spatial representations of actions. PMID- 15543555 TI - S19 ribosomal protein dimer augments metal-induced apoptosis in a mouse fibroblastic cell line by ligation of the C5a receptor. AB - To analyze the role of S19 ribosomal protein (RP S19) in apoptosis, murine NIH3T3 were transfected with either hemagglutinin peptide-tagged (HA) wild-type human RP S19 or a mutant (Gln137Asn) that is resistant to transglutaminase-catalyzed cross linked-dimerization. Transfection with the mutant HA-RP S19 inhibited manganese (II) (Mn II)-induced apoptosis whereas the wild-type HA-RP S19 augmented apoptosis and a mock transfection had no effect. Release of the wild-type HA-RP S19 dimer but not the mutant HA-RP S19 was observed during the apoptosis. The reduced rate of apoptosis of the cells transfected with the mutant HA-RP S19 was overcome by addition of extracellular wild-type RP S19 dimer. The apoptosis rates in cells transfected with either form of human HA-RP S19 and in mock transfectants were reduced to about 40% by the presence of anti-RP S19 antibody in the culture medium. Immunofluorescence staining and fluorescence-activated cell sorting (FACS) analysis showed that the cell surface expression of the receptor for cross-linked RP S19 dimer, C5a receptor, increased during apoptosis, concomitant with phosphatidylserine exposure. The expression of the C5a receptor gene also increased twofold. Apoptosis rates in the transfected and control cell lines were also reduced by the presence of an anti-mouse C5a receptor monoclonal antibody or of a peptide C5a receptor antagonist. These results indicated the presence of an RP S19 dimer- and C5a receptor-mediated autocrine-type augmentation mechanism during Mn II-induced apoptosis in the mouse fibroblastic cell line. In contrast to the RP S19 dimer, C5a actually inhibited apoptosis, suggesting that signaling through the C5a receptor varies depending on the ligand bound. PMID- 15543556 TI - Spatio-temporal dynamics at rDNA foci: global switching between DNA replication and transcription. AB - We have investigated the in situ organization of ribosomal gene (rDNA) transcription and replication in HeLa cells. Fluorescence in situ hybridization (FISH) revealed numerous rDNA foci in the nucleolus. Each rDNA focus corresponds to a higher order chromatin domain containing multiple ribosomal genes. Multi channel labeling experiments indicated that, in the majority of cells, all the rDNA foci were active in transcription as demonstrated by co-localization with signals to transcription and fibrillarin, a protein involved in ribosomal RNA processing. In some cells, however, a small portion of the rDNA foci did not overlap with signals to transcription and fibrillarin. Labeling for DNA replication revealed that those rDNA foci inactive in transcription were restricted to the S-phase of the cell cycle and were replicated predominantly from mid to late S-phase. Electron microscopic analysis localized the nucleolar transcription, replication, and fibrillarin signals to the dense fibrillar components of the nucleolus and at the borders of the fibrillar centers. We propose that the rDNA foci are the functional units for coordinating replication and transcription of the rRNA genes in space and time. This involves a global switching mechanism, active from mid to late S-phase, for turning off transcription and turning on replication at individual rDNA foci. Once all the rRNA genes at individual foci are replicated, these higher order chromatin domains are reprogrammed for transcription. PMID- 15543557 TI - RNA-dependent nuclear matrix contains a 33 kb globin full domain transcript as well as prosomes but no 26S proteasomes. AB - Previously, we have shown that in murine myoblasts prosomes are constituents of the nuclear matrix; a major part of the latter was found to be RNase sensitive. Here, we further define the RNA-dependent matrix in avian erythroblastosis virus (AEV) transformed erythroid cells in relation to its structure, presence of specific RNA, prosomes and/or proteasomes. These cells transcribe but do not express globin genes prior to induction. Electron micrographs show little difference in matrices treated with DNase alone or with both, DNase and RNase. In situ hybridization with alpha globin riboprobes shows that this matrix includes globin transcripts. Of particular interest is that, apparently, a nearly 35 kb long globin full domain transcript (FDT), including genes, intergenic regions and a large upstream domain is a part of the RNA-dependent nuclear matrix. The 23K type of prosomes, previously shown to be co-localized with globin transcripts in the nuclear RNA processing centers, were found all over the nuclear matrix. Other types of prosomes show different distributions in the intact cell but similar distribution patterns on the matrix. Globin transcripts and at least 80% of prosomes disappear from matrices upon RNase treatment. Interestingly, the 19S proteasome modulator complex is insensitive to RNase treatment. Only 20S prosomes but not 26S proteasomes are thus part of the RNA-dependent nuclear matrix. We suggest that giant pre-mRNA and FDTs in processing, aligning prosomes and other RNA-binding proteins are involved in the organization of the dynamic nuclear matrix. It is proposed that the putative function of RNA within the nuclear matrix and, thus, the nuclear dynamic architecture, might explain the giant size and complex organization of primary transcripts and their introns. PMID- 15543558 TI - Characterization of events associated with apoptosis/anoikis induced by snake venom metalloproteinase BaP1 on human endothelial cells. AB - Human endothelial EA.hy926 cells were incubated with BaP1, a hemorrhagic metalloproteinase purified from Bothrops asper snake venom. Since the first hour of incubation with the proteinase, cells started showing DNA fragmentation, detected by a terminal deoxynucleotidyl transferase-mediated dUDP nick-end labeling (TUNEL)-based photometric enzyme-linked immunosorbent assay (ELISA). At later times, DNA fragments were predominantly located outside the cells, evidencing plasma membrane rupture. DNA fragmentation was completely abolished by Batimastat, a potent inhibitor of metalloproteinase enzymatic activity. Apoptosis induced by BaP1 on endothelial cells was independent of two Bcl-2 family members (anti-apototic Bcl-xL and pro-apoptotic Bax), that did not show any changes in their expression during a 24 h-treatment period. Interestingly, IkappaBalpha, an inhibitor of NFkappaB, decreased after 24 h of treatment, suggesting further activation of the transcription factor. When some elements of the apoptotic extrinsic pathway were assessed, it was observed that procaspase-8 completely disappeared after 24 h of treatment with BaP1, probably indicating its activation by a death receptor, whereas caspase-8 inhibitor, cellular FLICE-inhibitory protein (cFLIP(L)), increased its expression since the first hours of BaP1 incubation. In conclusion, treatment of human endothelial cells with BaP1 induces apoptosis/anoikis, independently of Bcl-2 family members Bax and Bcl-xL and associated with caspase-8 activation and cFLIP(L) up-regulation. Apoptosis was completely dependent on BaP1 enzymatic activity. Similarities between this and other endothelial cell anoikis-related systems suggest that BaP1 and other snake venom metalloproteinases may be useful experimental tools in the study of death related events that occur when adherent cells loose contact with extracellular matrix. PMID- 15543559 TI - Adenylyl cyclase-cAMP system inhibits thrombin-induced HSP27 in vascular smooth muscle cells. AB - We previously reported that thrombin stimulates the induction of heat shock protein (HSP) 27 via p38 mitogen-activated protein (MAP) kinase activation in aortic smooth muscle A10 cells. In the present study, we investigated the effect of the adenylyl cyclase-cAMP system on the thrombin-stimulated induction of HSP27 in A10 cells. Forskolin, a direct activator of adenylyl cyclase, reduced the thrombin-induced p38 MAP kinase phosphorylation, and significantly suppressed the thrombin-stimulated accumulation of HSP27. However, dideoxyforskolin, a forskolin derivative that does not activate cAMP, failed to suppress the HSP27 accumulation. Furthermore, dibutyryl-cAMP (DBcAMP), a permeable analog of cAMP, significantly suppressed the accumulation of HSP27. On the other hand, calphostin C, an inhibitor of protein kinase C (PKC), reduced the thrombin-induced p38 MAP kinase phosphorylation, and significantly suppressed the thrombin-stimulated accumulation of HSP27. Moreover, forskolin reduced the p38 MAP kinase phosphorylation induced by the 12-O-tetradecanoylphorbol-13-acetate (TPA), a PKC activating phorbol ester, and significantly suppressed the TPA-stimulated accumulation of HSP27. These results indicate that adenylyl cyclase-cAMP system has an inhibitory role in thrombin-stimulated HSP27 induction in aortic smooth muscle cells, and the effect seems to be exerted on the thrombin-induced PKC- p38 MAP kinase signaling pathway. PMID- 15543560 TI - Deregulation of DNA methyltransferases and loss of parental methylation at the insulin-like growth factor II (Igf2)/H19 loci in p53 knockout mice prior to tumor development. AB - To ascertain whether p53 deficiency in vivo leads to the deregulation of DNA methylation machinery prior to tumor development, we investigated the expression profile of DNA methyltransferases in the thymus and the liver of p53(+/+), p53(+/ ), and p53(-/-) mice at 7 weeks of age before tumor development. The expression of DNA methyltransferases was examined in the thymus at 7 weeks of age, since the malignant T-cell lymphoma develops most frequently in p53(-/-) mice around 20 weeks of age. Both mRNA and protein levels of Dnmt1 and Dnmt3b were increased in the thymus and the liver of p53-deficient mice. The expression of Dnmt3a was also increased in the liver but not in the thymus of p53-deficient mice. Dnmt3L expression was reduced in the thymus of p53(+/-) and p53(-/-) mice. The total 5 methylcytosine (5-MeC) in the genomic DNA of p53(+/+), p53(+/-), and p53(-/-) mice was quantitated by dot-blot using antibody against 5-MeC. Global methylation was increased in the thymus and the liver of p53-deficient mice. To correlate the deregulated expression of DNA methyltransferases with the disturbance of the epigenetic integrity, we examined the DNA methylation of the imprinting control region (ICR) at the insulin-like growth factor II (Igf2)/H19 loci in the thymus and the liver of p53(+/+), p53(+/-), and p53(-/-) mice. The region containing two CCCTC binding factor (CTCF) binding sites in the 5'-ICR tended to be hypomethylated in the thymus of p53(-/-) mice, but not in the liver. The expression profile of Igf2 and H19 indicated that the thymus-specific changes of Igf2 and H19 expression were coherent to the hypomethylation of the ICR in the thymus. Our results suggest that p53 is required for the maintenance of DNA methylation patterns in vivo. PMID- 15543561 TI - Recommended dietary allowance for vitamin C in the United States is also applicable to a population of young Japanese women. AB - The recommended dietary allowance (RDA) for ascorbic acid (AA) in Canada and the United States has been set for several years at 75 mg/day for women 19-30 years old. Recently this level was questioned, and an increase to 90 mg/day was suggested. For Japanese women in the same age group, we found that the RDA for AA is currently 100 mg/day. Our goal was to determine which RDA is sufficient for maintaining a serum concentration of AA in young Japanese women above the lower reference limit of 7.0 mg/L. We measured serum AA concentrations by an ascorbate oxidase method in 176 healthy Japanese women (19-26 years old). We also performed an ROC analysis to estimate the optimal cutoff value for oral dosage to distinguish individuals with hypovitaminosis-C (<7.0 mg/L) from those with a normal serum AA. We evaluated the Japanese RDA using the 75 or 90 mg/day U.S. RDA and the weight ratio between Japanese and U.S. women, and discovered that the RDA value ranged between 66 and 79 mg/day. From the ROC analysis, we found that the optimal daily dosage of AA is approximately 75 mg/day. This value gave the highest efficiency, sensitivity, negative predictive value, and positive likelihood ratio, and the lowest negative likelihood ratio. Therefore, an RDA of 100 mg/day may be unnecessarily high for young Japanese women. PMID- 15543562 TI - Peroxisome proliferator-activated receptor gamma gene polymorphism is associated with serum triglyceride levels and body mass index in Japanese type 2 diabetic patients. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) controls adipocyte differentiation and regulates lipid and glucose homeostasis. Therefore, the PPARgamma gene may affect insulin sensitivity and resistance. We analyzed the relationship between C/T exon 6 polymorphism of the PPARgamma gene and various clinical parameters in type 2 diabetic patients. There were no significant differences in the frequencies of genotype and allele between diabetic patients with and without nephropathy. Diabetic patients were divided into two groups: patients bearing at least one T allele (CT/TT), and patients with no T allele (CC). Levels of serum triglyceride and body mass index (BMI) were significantly higher in the CT/TT genotype group than in the CC genotype group. Since obesity affects insulin resistance, the diabetic patients were also divided into two groups: those with a BMI of <23, and those with a BMI of >23. In patients with a BMI of <23, there was no significant change in the levels of glycosylated hemoglobin A1c (HbA1c) between the CC and CT/TT genotype groups. However, in patients with a BMI of >23, HbA1c levels were significantly higher in the CT/TT genotype group than in the CC genotype group. It appears that the CT/TT genotype with PPARgamma gene polymorphism may contribute to higher BMI and higher serum triglyceride and HbA1c levels in Japanese type 2 diabetic patients. PMID- 15543563 TI - Five polymorphisms in gene candidates for cardiovascular disease in Afro Brazilian individuals. AB - Associations of polymorphisms in the angiotensin I-converting enzyme (ACE), apolipoprotein B (APOB) and apolipoprotein E (APOE) genes with hypertension and variations in lipid serum levels were evaluated in 184 Afro-Brazilians with a familial history of coronary artery disease (CAD). ACE (Ins/Del) and APOB (Ins/Del, XbaI, and EcoRI) and APOE (HhaI) polymorphisms were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses on agarose, and polyacrylamide gel electrophoresis. Serum lipids were measured by means of routine enzymatic assays. The results showed a high frequency of hypertension (44%) in Afro-Brazilians that was increased in subjects >40 years old and those with a blood mass index (BMI) higher than 25 kg/m(2) (P<0.001). The ACE Del allele was associated with hypertension in men >40 years old (P<0.05). APOE (HhaI) and APOB (XbaI and Ins/Del) polymorphisms were not associated with hypertension or variations in serum concentrations of lipids, while subjects with the APOB E- allele had higher low-density lipoprotein cholesterol (LDL-C) levels than E+ carriers (P<0.05). These results suggest that ACE Ins/Del polymorphism is associated with hypertension, and APOB EcoRI polymorphism is associated with LDL-C variation in Afro-Brazilians. PMID- 15543564 TI - Retrospective study on poliomyelitis cases in northern Greece. AB - Here we report the results of a retrospective study on the epidemiological characteristics and genetic relationships of the virus isolates responsible for the last poliomyelitis cases in Greece. The last wild poliomyelitis case in Greece was detected in 1996, and the last vaccine-related strain was isolated in 1998. The whole of Europe, including Greece, is now considered to be polio-free. PMID- 15543565 TI - Sensitive noninvasive marker for the diagnosis of probable bacterial or viral infection. AB - Urinary trypsin inhibitor (uTi) is a product of elastase-mediated degradation of interleukin-alpha-inhibitor (I-alpha-I). Its activity increases in the urine of patients with a malignancy, inflammation, or infection, or in late pregnancy. The objective of this study was to compare the sensitivity of uTi in urine with that of serum quantitative C-reactive protein (CRP) for diagnosing infection, as indicated by white cell response and clinical assessment. Ninety controls and 171 patients with various systemic infections were enrolled. We measured uTi enzymatically on a Cobas Fara (Roche Diagnostics). Patients were separated into bacterial, probable bacterial, viral, or probable viral groups based on the results of a complete blood count with differential (CBC), urinalysis (UA), and clinical assessment. In the bacterial (n=70) and control (n=90) groups, the uTi values (mean+/-SE) were 25.3+/-3.1 mg/L and 2.8+/-0.8 mg/L, respectively. uTi (at 2.7 mg/L) had a diagnostic sensitivity of 91% and specificity of 82% (AUC=0.889), whereas CRP (at a cutoff of 10 mg/L) had a sensitivity and specificity of 82% and 96%, respectively (AUC=0.921). As a marker of infection (positive in both bacterial and viral groups), uTi had a sensitivity of 91% (AUC=0.884) vs. 89% (AUC=0.828) for CRP. Our data indicate that uTi has sufficient clinical sensitivity for screening systemic infections, and may have diagnostic value as a noninvasive test. PMID- 15543566 TI - Insulin-like growth factors in patients with liver cysts. AB - Insulin-like growth factors (IGFs) play an important role in cell growth and differentiation, and the liver is the main source of IGFs and IGF-binding proteins (IGFBPs) that appear in the circulation. The effect of liver cysts on the circulating IGF system was studied in this work. Serum concentrations of IGF I and -II were measured by radioimmunoassay, IGFBP patterns were characterised by ligand-affinity and immunoblotting, and a lectin-binding assay was used to investigate the glyco component of IGFBP-3 complexes. IGF-I and -II concentrations in patients with cysts were significantly lower compared to those in healthy individuals (P<0.0001 and P<0.01, respectively), and the decrease was related to age but not sex. The overall mean concentrations of IGF-I and -II were not significantly different whether the cysts were caused by Echinococcus granulosus, cross-reactive pathologies, or some other factor. IGFBP profiles correlated with the amount of IGF present: patients with lower IGF-I concentrations expressed decreased IGFBP-3 and elevated IGFBP-2 levels. Increased IGFBP-3 proteolytic activity in the patients' blood was not detected by immunoblotting. In the lectin-binding assay, IGFBP-3 complexes in the circulation of patients demonstrated reactivity similar to that in healthy persons, suggesting that the overall structure of the saccharide moieties of the IGFBP-3 complexes was not significantly altered due to liver cyst formation. PMID- 15543567 TI - Changing epidemiology of rotavirus G-types circulating in Hong Kong, China. AB - Group A rotaviruses are the most common cause of severe diarrhoeal disease in young children worldwide. The development of a vaccine is advocated by the World Health Organization. Obtaining local baseline information regarding rotavirus strain variation is important to ensure matching of circulating and vaccine strains. The current study was undertaken to determine the epidemiology of rotavirus G-types in Hong Kong in anticipation of a vaccination program. From 2001 to 2002 over a period of one year, diarrhoeal stool specimens known to be positive for rotavirus were subjected to G-typing by reverse transcriptase polymerase chain reaction using nested type-specific primers. Rotavirus G-type distribution was correlated with patient demographics. Among 747 rotavirus positive stool specimens, 723 strains could be G-typed as G1 (302, 40.4%), G2 (128, 17.1%), G3 (231, 30.9%), G4 (24, 3.2%), and G9 (38, 5.1%). G1 strains were found predominantly in those 5 years old or younger (P < 0.0001), while G2 strains were more prevalent among those over 5 years of age (P < 0.001). When compared with similar studies in 1983 to 1984 and 1999 to 2000, there were significant changes in the prevalence of various G-types, with consistent detection of G9 strains in the current study. It is concluded that rotavirus G type distribution in Hong Kong has varied with time. Continuous monitoring of the epidemiology of rotavirus is important, especially in anticipation of the introduction of a vaccine, in order to document its impact and to ensure its continued effectiveness. PMID- 15543568 TI - Do Bcl-2 and survivin help distinguish benign from malignant B-cell lymphoid aggregates in bone marrow biopsies? AB - Bcl-2 and survivin are cellular proteins that are known to be inhibitors of apoptosis and are commonly found in malignant tissues, including lymphomas. In previous studies, it has been shown that staining for bcl-2 can help distinguish between benign and malignant lymphoid aggregates in bone marrow biopsies. To determine whether staining for survivin expression in lymphoid aggregates can aid investigators in making this clinically important distinction, we stained bone marrow biopsies from 10 patients with benign lymphoid aggregates, and 15 malignant ones derived from B cells (six mantle cell, four follicular cells, two diffuse large cell, two small lymphocytic cell, and one marginal zone lymphoma) with antibodies to CD3, CD20, bcl-2, and survivin by an indirect immunoperoxidase technique. Whereas staining for bcl-2 was significantly stronger in the malignant lymphoid aggregates (P=0.001), both the control and malignant cases were almost uniformly negative for survivin expression. Only three cases (two mantle cell and one small lymphocytic lymphoma) showed very faint expression of survivin. Although bcl-2 and survivin both act to inhibit apoptosis, their expressions do not parallel each other. Survivin is not significantly expressed in either benign or malignant bone marrow aggregates, and therefore measuring its expression does not help distinguish benign from malignant B-cell bone marrow lymphoid aggregates. PMID- 15543569 TI - Heparan sulfate proteoglycans interact exclusively with conformationally intact HPV L1 assemblies: basis for a virus-like particle ELISA. AB - In this article, we demonstrate that interaction of human papillomavirus-like particles (HPV-VLPs) with the putative glucosaminoglycan binding receptor is strictly dependent on conformational integrity. Such conformations are present on VLPs and capsomeres but not on monomers of the major capsid protein, L1, confirming reports that capsomeres can induce virus-neutralizing antibodies. Furthermore, we show the suitability of this specific interaction for development of VLP-based enzyme-linked immunosorbent assays (ELISAs), using heparin for indirect coupling of VLPs to microtiter plates, which may add an intrinsic quality control. This avoids presentation of linear, often highly cross-reactive epitopes of L1. In addition, heparin specifically interacts with a wide variety of HPV types, making it a prime candidate for a universal capture molecule. PMID- 15543570 TI - Identification of epitopes in the nucleocapsid protein of Nipah virus using a linear phage-displayed random peptide library. AB - A random peptide library of heptamers displayed on the surface of M13 bacteriophage was used to identify specific epitopes of antibodies in pooled sera of swine naturally infected by Nipah virus. The selected heptapeptides were aligned with protein sequences of Nipah virus and several putative epitopes were identified within the nucleocapsid protein. A total of 41 of 60 (68%) selected phage clones had inserts resembling a region with the sequence SNRTQGE, located at the C-terminal end (amino acids 503-509) of the nucleocapsid protein. The binding of antibodies in the swine and human antisera to the phage clone was inhibited by a synthetic peptide corresponding to this region. Epitopes identified by phage display are consistent with the predicted antigenic sites for the Nipah virus nucleocapsid protein. The selected phage clone used as a coating antigen discriminated swine and human Nipah virus sera-positive from sera negative samples exhibiting characteristics, which might be attractive for diagnostic tests. PMID- 15543571 TI - Epstein-Barr viral load as a marker of lymphoma in AIDS patients. AB - Epstein-Barr virus (EBV) is implicated in the pathogenesis of acquired immunodeficiency syndrome (AIDS) lymphoma, and viral DNA is present within the malignant cells in about half of affected patients. We examined the extent to which EBV viral load is elevated in the plasma of AIDS lymphoma patients compared to AIDS patients with opportunistic infections. Sixty-one AIDS patients were studied including 35 with lymphoma (24 non-Hodgkin, six Hodgkin, and five brain lymphoma) and 26 with various opportunistic infections. In situ hybridization revealed EBV encoded RNA (EBER) expression in the malignant cells of 17/28 AIDS lymphomas (61%). In 232 serial plasma samples from 35 lymphoma patients and in 128 samples from AIDS controls, EBV viral load was assayed by quantitative polymerase chain reaction (Q-PCR) using a TaqMan probe targeting the BamH1W sequence. EBV was detected in plasma from all 17 EBER-positive AIDS lymphoma patients, with viral loads ranging from 34 to 1,500,000 copies per ml (median 3,210). Viral load usually fell rapidly upon initiation of lymphoma therapy and remained undetectable except in two patients with persistent tumor. In 11 AIDS patients, whose lymphoma lacked EBER expression, and in 26 control patients without lymphoma, levels of EBV in plasma were usually low or undetectable (range 0-1,995 and 0-2,409, median 0 and 0, respectively). There was no association between EBV viral load and human immunodeficiency virus (HIV) load or CD4 count. In conclusion, EBV viral load shows promise as a tool to assist in diagnosis and management of EBV-related lymphoma patients. PMID- 15543572 TI - In vitro and in vivo analysis of human herpesvirus-6 U90 protein expression. AB - In order to establish a reliable method for the detection of human herpesvirus-6 (HHV-6) B antigens in peripheral blood mononuclear cells (PBMCs) collected from HHV-6 infected patients, we created a polyclonal antibody against the HHV-6 B U90 protein (IEA/ex3) and used it to examine the expression of this protein in virus infected cells and patients' PBMCs. This antibody reacted with 170 and 195 kDa proteins in HHV-6 B-infected cord blood mononuclear cells. The IEA/ex3 antigen was detected in cord blood mononuclear cells at 6 hr post-infection, and the number of infected cells reached its maximum at 48 hr post-infection. The antigen stained in a punctate pattern and partially localized to the promyelocytic leukemia (PML) protein body. We also examined 60 PBMC samples from 60 febrile children (3-19 months old) and detected IEA/ex3 antigen in the PBMCs by laser scanning microscopy. HHV-6 was isolated from 31 of the 60 samples. The sensitivity and specificity of the antigen detection were 84% (26/31) and 97% (28/29), respectively, in the samples with virus detected. The mean number of antigen-positive PBMCs was 409/10(6) cells in 20 samples with viral isolation. A significant correlation (r = 0.566; P = 0.008) was observed between the viral load and number of antigen-positive cells. Although IEA/ex3 antigen was detected by laser-scanning microscopy in PBMCs (without cultivation) collected from six patients with isolated virus, it was detected in only one sample by conventional fluorescence microscopy. Increasing the intensity by cultivation (24 hr) resulted in a higher detection rate (5/6) even by conventional fluorescence microscopy, which is available in most hospital laboratories. PMID- 15543573 TI - Phylogenetic analysis of hepatitis A virus in Thailand. AB - Human hepatitis A virus (HAV) is a major causative agent of acute hepatitis, and the isolates are categorised into four genotypes: I (GI), II (GII), III (GIII), and VII (GVII). Although viral hepatitis has been detected under a nationwide surveillance system in Thailand, the genetic variation of HAV has not yet been determined. In the present study, serum specimens were collected from acute hepatitis patients in Thailand from 1998-2002. The IgM-class antibody to HAV was detected in 156 out of 394 sera, counting as many as 39.6% of acute viral hepatitis cases. HAV RNA in the serum specimens was amplified by reverse transcription polymerase chain reaction (RT-PCR), and a phylogenetic analysis of the putative VP1/2A junction of the genome was performed. The isolates were grouped into two genetic groups, GIA and GIB. This is the first report to identify subgenotype IB (GIB) in Thailand. The genetic segregation was closely related to the province where hepatitis A occurred and the serum specimens were collected. In addition, genetically similar strains were identified in both 1998 and 2001-2002 isolates from two close provinces in the southern part of Thailand, suggesting that a strain indigenous to the province or district has been circulating in southern Thailand. PMID- 15543574 TI - Comparison of full length sequences of hepatitis B virus isolates in hepatocellular carcinoma patients and asymptomatic carriers of Korea. AB - Relatively few genomic sequences of Korean hepatitis B virus (HBV) isolates are available. Moreover, no comparative study has been made between the full-length genomes of Korean HBV isolates and clinical status. To evaluate mutations in HBV isolates obtained from chronically infected HBV patients in terms of clinical significance, we determined the genomic sequences of HBV isolates obtained from three hepatocellular carcinoma (HCC) patients (He52, He53, and He82) and from three asymptomatic carriers (He74, He100, and He127). A comparison of sequence variations showed that the HBV isolates from the three HCC patients showed higher frequencies of mutation than the isolates from the three asymptomatic carriers. Three characteristic mutation patterns were identified in the HBV isolates from the HCC patients, which distinguished the HBV isolates from the asymptomatic carriers. First, HBV isolates from the three HCC patients both had double mutations in a core promoter (T1762/A1764) and a precore mutation (A1896). Second, although these isolates belonged to genotype C, 11 amino acids deletions in the preS1 region, specific for HBV genotype D, were detected in the isolates of two HCC patients (He52 and He82). Third, mutations (I127T/N, K130M, and V131I) at three codons in the carboxy functional region of X protein were observed in isolates from all three HCC patients. Additionally, phylogenetic analysis based on the entire HBV sequences showed that all six isolates belonged to genotype C2, as do other Korean strains. PMID- 15543575 TI - Recurrent high level parvovirus B19/genotype 2 viremia in a renal transplant recipient analyzed by real-time PCR for simultaneous detection of genotypes 1 to 3. AB - Organ transplant recipients infected with parvovirus B19 frequently develop persistent viremia associated with chronic anemia and pure red cell aplasia. In this study, a male renal transplant recipient who had been infected with parvovirus B19/genotype 2 after renal transplantation at the age of 34 years is described. The patient was repeatedly treated with high dose intravenous immunoglobulin (IVIG) that resulted in the resolvement of symptoms but not in virus eradication. During an observation period of 33 months after transplantation three phases associated with high parvovirus B19 viremia were observed. Both the first and the second viremic phases were combined with severe anemia. Parvovirus B19 specific IgM-antibodies were initially detected at the beginning of the second phase in continually rising concentrations. Initially eradication of the virus by immunoglobulin therapy was reported after the first viremic phase [Liefeldt et al. (2002): Nephrol Dial Transplant 17:1840-1842]. Retrospectively this statement has to be corrected. It was based on the use of a qualitative PCR assay specific for parvovirus B19 genotype 1 associated with reduced sensitivity for detection of genotype 2. After sequence analysis of the viral DNA and adjustment of a real-time PCR assay (TaqMan) for quantitative detection of all three B19 virus genotypes analysis of consecutive serum samples allowed the demonstration of long lasting phases with reduced viral loads following IVIG-treatment. These results demonstrate that IVIG treatment of parvovirus B19-triggered anemia in transplant recipients offers an opportunity to resolve symptoms, but does not guarantee eradication of the virus. Since reactivation of parvovirus B19 infection can result in high virus load associated with the recurrence of symptoms repeated screening for viral DNA is recommended using the TaqMan system established for quantitative detection of all three genotypes of parvovirus B19. PMID- 15543576 TI - An evaluation of single nucleotide polymorphisms used to differentiate vaccine and wild type strains of varicella-zoster virus. AB - Rashes following immunization with the vaccine strain (vOka) of varicella-zoster virus (VZV) may occur in up to 5% of children and 10% of adults. In 40% of cases, the causative virus is the vaccine strain and in 60% wild type virus is found. Several reports have identified three restriction site polymorphisms in ORF 62 and the loss of one in ORF 6, which differentiate vOka from wild type VZV, including the parental wild type strain from which vOka, is derived. Using polymerase chain reaction (PCR), restriction enzyme analysis, and sequencing, we analyzed the presence of these markers in the GlaxoSmithKline (GSK, UK) and Merck vaccine preparations as well as in 15 vaccine virus rashes and 15 wild type UK viruses. Our data suggest that a Sma1 positive and an Nae1 positive site in ORF 62 are present in the GSK and Merck vaccine preparations and all vaccine virus rashes. By contrast, a BssHII positive vaccine virus restriction site in ORF 62 and an Alu1 negative site in ORF 6 were mixed in the GSK and Merck vaccines and absent in some of the vaccine rashes. The BssHII site was also present in the European wild type C viruses in UK. The data suggest that unlike the Biken vaccine preparation, the Merck and GSK vaccine preparations are polymorphic for the BssHII and Alu1 restriction sites. These sites are also present variably in the vaccine viruses causing rashes following vaccination, and are therefore unreliable markers for differentiating vOka and wild type VZV strains. PMID- 15543577 TI - Measles virus genotype circulation in Spain after implementation of the national measles elimination plan 2001-2003. AB - Molecular characterization of measles virus is important for disease surveillance and for monitoring elimination of the virus throughout the world. Furthermore, knowledge of genotype distribution in as many countries as possible, is useful for tracing the origin of a strain, especially in countries without endemic measles disease, where most cases are imported. Data on genotypes circulating in Spain from 1970 to 1997 showed the prevalence of genotypes C1, C2, and D6, with subsequent replacement of each other. After the establishment of the Spanish Measles Elimination Plan, genotyping with a new retrotranscriptase polymerase chain reaction (RT-PCR) was undertaken directly on 92 specimens, corresponding to 90 patients, which were positive for measles by a different diagnostic RT-PCR. Genotypes B3, D4, D8, A, C2, H1, and D7 were found in different autonomous communities (Madrid, Balearic Islands, Valencia Community, Extremadura, Andalusia, Canary Islands and Murcia) between 2001 and 2003 with none of these genotypes being prevalent. After the introduction of the vaccine in 1978, the incidence of the disease decreased from 150,000 cases in 1977 to 64 in 2002. This could be the reason for the change observed in the pattern of measles genotype circulation, since this pattern was reported in countries at an advanced stage of eradication of measles. This report considers that Spain is on the way to eradicating measles. PMID- 15543578 TI - Histone acetyltransferase activity of p300 enhances the activation of IL-18 promoter. AB - Interleukin-18 (IL-18), an important regulator of innate and acquired immune responses expressed from a variety of cell types, is a pleiotropic cytokine in the development of T helper type 1 (Thl) cells. The p300/CBP (CREB-binding protein) coactivator proteins are important histone acetyltransferases (HATs) that regulate the transcription of many genes. Whether p300/CBP play a role in the IL-18 expression has not been investigated previously. In this study, we analyzed the roles of p300 in the regulation of mouse IL-18 by using RT-PCR and a series of co-transfection studies. We showed that p300 had a stimulating effect on the endogenous IL-18 mRNA synthesis and on the activity of IL-18 p1 promoter. The results also showed that IL-18 p1 promoter activity was enhanced by p300 in a dose-dependent manner. Moreover, the p300-mediated activation function can be suppressed by the adenovirus E1A protein, which inhibits the HAT function of p300. Also, a mutation in p300 HAT region abolished the effect of p300 on IL-18 activation. These data further indicate that the acetylase activity of p300 was indispensable to its function. Furthermore, we found that p300 was able to enhance the effect of the transcription factor c-Fos on activation of the IL-18 promoter. Data presented in this paper implicate important roles of p300 in the transcriptional regulation of IL-18. PMID- 15543579 TI - Prevalence of human papillomavirus genotypes in women from three clinical settings. AB - Prevalence of 27 human papillomavirus (HPV) genotypes was assessed in 1,331 women in three clinical settings: Family planning clinic (low-risk HIV-, n = 202, 21.3% HPV+), colposcopy clinic (high-risk HIV-, n = 854, 34.3% HPV+), and HIV outpatient clinic (HIV+, n = 275, 48.7% HPV+). Compared to women from both family planning and colposcopy clinics, HIV+ women revealed significantly higher prevalence of infection with oncogenic, non-oncogenic, and multiple HPV types. HPV types 52 and 51 were most prevalent in the low-risk HIV- women, whereas in the high-risk HIV- women, HPV types 16, 52, 58, and 35 were most prevalent. Interestingly, in the HIV+ women, less characterized types 83, 53, and 54 were most prevalent. The distinct profiles of genotype prevalence persisted after stratification by Pap smear status. After adjustment for concurrent infections with other types, HPV type 51 in the low-risk HIV- women, and types 16, 35, 39, 45, 52, and 58 in the high-risk HIV- women were significantly associated with cytologic abnormalities (exact P < 0.05). In HIV+ women across CD4 cell count strata, HPV types 42, 16, and 82 revealed significant decreasing trends with increasing CD4 counts (exact P for trend < 0.05). These data suggest distinct genotypic prevalence profiles in women at diverse risk for cervical cancer. The association of several genotypes with cytologic abnormalities underscores the need for vaccines targeting a wide range of HPV types. PMID- 15543580 TI - Laboratory diagnosis and surveillance of human respiratory viruses by PCR in Victoria, Australia, 2002-2003. AB - Respiratory viruses were identified by the polymerase chain reaction (PCR) in more than 4,200 specimens collected during 2002 and 2003 in Victoria, Australia from patients admitted to hospitals or participating in an influenza surveillance program. Influenza viruses and picornaviruses were important causes of morbidity in both years. Additional testing of picornavirus-positive samples suggested that rhinoviruses but not enteroviruses were more likely to be associated with respiratory symptoms, irrespective of the season in which they circulated. Detection of influenza viruses was strongly associated with the clinical symptoms of cough, fever, and fatigue; but each of the other respiratory viruses occasionally caused these symptoms or was responsible for symptoms severe enough to require hospitalization. Human coronaviruses HCoV-OC43 and HCoV-229E circulated at low levels throughout the study period with peak activity in winter, but overall did not circulate as widely as has often been reported for these agents. Evidence for the human metapneumovirus (hMPV) was only sought in the second year of the study and revealed low-level circulation of this virus, mainly in the cooler months among the very young and adult populations. The detection rate of all viruses declined with increasing age of the patient, particularly in hospital patients. Infection with more than one respiratory virus occurred in a small number of patients; picornaviruses were most commonly implicated in these dual infections. PMID- 15543581 TI - Quantitative analysis of human herpesvirus 6 cell tropism. AB - Although HHV-6A and -6B are known to replicate preferably in human T-lymphocytes, in vitro infection of several other cell types has been described. Also, the finding that both variants use the ubiquitous molecule CD46 as a membrane receptor fully supports the possibility of a broad cellular tropism. However, productive infection, which requires complete progression through the viral replication cycle, depends on multiple cellular processes. Our studies were aimed at determining the differences in replication efficiency according to the cell type infected and at relating these differences to the sequential transcriptional events preceding DNA replication. A strong expression of immediate-early, early, and late genes was only seen in the T-lymphoblastoma lines. In the other cell lines, there was no clear correlation between the level of transcription and the final outcome of replication. Finally, we investigated the cytopathic effects of HHV-6 on different cell types of neural origin (oligodendrocytes, astrocytes, and neurons) in greater detail, and found that although all three sustained HHV-6 replication, HHV-6A was more neurovirulent than HHV-6B. This was confirmed in primary human oligodendrocyte cultures. PMID- 15543582 TI - Epidemiological characteristics of human herpesvirus-8 infection in a large population of antenatal women in Zambia. AB - Comprehensive data describing epidemiological characteristics of the human herpesvirus-8 or Kaposi's sarcoma-associated herpesvirus (HHV-8 or KSHV) infection among pregnant women in a central sub-Saharan Africa are not available. This study determined virus prevalence estimates and the risk factors associated with HHV-8 infection. Cross-sectional, enrollment visit data were analyzed from a prospective cohort study of perinatal transmission of HHV-8 in Lusaka, Zambia. Exposure data were obtained via structured interview, physical examination, medical chart review, and laboratory testing. Among 3,160 antenatal women serologically screened for HHV-8 between September 1998 and October 2000, 40.2% were seropositive. The HHV-8 positive women were more likely to be co-infected with HIV-1 than those who were HHV-8 negative (34% vs. 26%; P < 0.0001). Of 154 variables evaluated by logistic regression analyses, only three risk factors, have emerged as independent predictors of HHV-8 positive serology: diagnosis of genital warts, HIV-1 co-infection and primary education. The association of HHV-8 infection with genital warts and HIV-1 co-infection suggests heterosexual transmission of HHV-8. HIV-1 infection may also act as a marker for particular behaviors, which could be sexual in nature, that are associated with both HIV-1 and HHV-8 transmission. Since HHV-8 facilitates development of AIDS-related Kaposi's sarcoma (KS), the results of this study could be utilized to identify specific population groups of pregnant women who are at increased risk for this disease. PMID- 15543583 TI - Serum Epstein-Barr virus DNA load in primary Epstein-Barr virus infection. AB - Specific viral laboratory diagnosis of primary Epstein-Barr Virus (EBV) infection is usually based on antibody-detection assays. During acute, lytic phase of infection, viral DNA can also be detected in serum. In the present study, the diagnostic utility of EBV DNA detection and quantitation in serum in primary EBV infection was investigated. The level of EBV DNA in the serum of 98 immunocompetent patients aged 1-47 years with symptomatic, antibody-confirmed EBV primary infection was assessed using a quantitative real-time PCR assay. The association between viral load and time after onset of disease, age and clinical and laboratory data was investigated. Quantitative PCR detected EBV DNA in 93 of 98 samples (94.9%), and the measured viral loads ranged from 3.8 x 10(1) to 6.6 x 10(4) copies/ml. EBV DNA detection exhibited a sensitivity of 94.9% and a specificity of 97.4% for primary EBV infection. EBV DNA was always detectable until day 12 after onset of symptoms, whereas no further positive PCR results were found after a period of 22 days after onset of disease. Detection of EBV DNA also showed a clearer association with the clinical manifestation of disease than the presence of EBV specific VCA IgG antibodies of low avidity. EBV DNA load was found to correlate inversely with the time after onset of disease (P < 0.001), and higher viral load levels were detected in younger (P = 0.009) and in hospitalized patients (P = 0.038). The results indicate that real-time PCR is a reliable tool for diagnosis of primary EBV infection early in the course of disease. In addition, EBV DNA detection may serve as a useful diagnostic supplement in serologically indeterminate EBV infections. PMID- 15543584 TI - HHV-6 and 7 DNA loads in lung tissues collected from patients with interstitial pneumonia. AB - The aim of this study is to determine whether human herpesvirus 6 (HHV-6) and HHV 7 might play an important role in causing interstitial pneumonia in patients who have not undergone transplantation. HHV-6 and HHV-7 DNAs were quantitated by real time polymerase chain reaction (PCR) in paraffin embedded lung tissues collected from 24 patients having the disease. Control tissues (without fibrosis) were also collected from 19 of the 24 patients. Statistical analysis was carried out by the Wilcoxon signed rank test or the Mann-Whitney U-test. HHV-6 DNA was detected in 3 (12.5%) of the 24 target tissues and 3 (15.8%) of the 19 control tissues, respectively. In contrast, HHV-7 DNA was detected in 19 (79.2%) of the 24 target tissues and 11 (57.9%) of the 19 control tissues. Neither HHV-6 DNA load (P = 0.6395) nor HHV-7 DNA load (P = 0.5966) in target tissues differed between males and females. Neither HHV-6 DNA load (P = 0.9589) nor HHV-7 DNA load (P = 0.7419) in target tissues differed between cases with and without underlying collagen disease. While HHV-6 DNA load did not differ between the target and control tissues (P > 0.9999), the HHV-7 DNA load was significantly higher in the target tissue than in the control tissue (P = 0.0298). This study suggests that HHV-7 may play an important role in causing interstitial pneumonia in patients who are not transplant recipients. PMID- 15543585 TI - Mode of HCV infection examined by polymorphism of hypervariable region-1 in cases of acute hepatitis C after accidental exposure to blood-borne pathogens. AB - Acute hepatitis C is known to respond better to interferon therapy than chronic hepatitis C. The reason for this difference remains unclear. The present study was undertaken to examine HCV quasispecies in blood from patients with acute hepatitis C caused by accidental exposure to blood-borne pathogens and in blood from the source patients. Three patients who developed hepatitis C (recipient patients; R-Pt.) and two patients who served as a source of HCV infection (source patients; S-Pt.) were the subjects of this study. The number of quasispecies and the genetic diversity in hypervariable region-1 (HVR-1) were examined on the basis of fluorescence single-strand conformation polymorphism and sequence analysis (FSSA). On the day of the accident, the number of quasispecies and genetic diversity were 13 and 36 in S-Pt.1 and 6 and 20 in S-Pt.3, respectively. At the time of diagnosis of acute hepatitis, the number of quasispecies and nucleotide diversity were 2 and 2 in R-Pt.1, 2 and 0 in R-Pt.2, and 4 and 0 in R Pt.3, respectively. Immediately before the start of treatment, the number of quasispecies and genetic diversity were 4 and 4 in R-Pt.1, 2 and 0 in R-Pt.2, and 3 and 0 in R-Pt.3., respectively. In three R-Pts, interferon therapy resulted in eradication of HCV. These findings indicate that in the early stage of HCV infection, only a portion of HCV transmitted from S-Pts to R-Pts can proliferate. The low number of quasispecies of HCV appears to be one of the reasons why acute hepatitis responds well to interferon therapy. PMID- 15543586 TI - Stability of human cytomegalovirus genotypes in persistently infected renal transplant recipients. AB - Although most genes are highly conserved among strains of human cytomegalovirus (HCMV), several are unusually variable. By analyzing the sequence of two variable genes (UL146 and UL74) amplified directly from whole blood DNA extracts, multiple HCMV strains were detected in blood samples from 5/11 virus-infected renal transplant patients. These five patients were seronegative prior to transplantation and their likely acquisition of the virus was via the donated organ; HCMV-positive immunocompetent donors may thus be capable of harboring and transmitting multiple virus genotypes. In sequential samples taken up to 140 days post transplant no mutations in either gene were detected from 9/10 patients, and in the remaining patient a single nucleotide change was detected in UL146, and none in UL74. All sequences grouped into previously defined genotypes, with the detection of multiple members of the UL74 group 5 genotype establishing this previously tentative genotype. Additionally, identical sequences were identified in viruses from different patients, including examples from geographically distinct regions. Thus, although UL146 and UL74 exhibit impressive variation among strains, their sequences are maintained stably within and between infected individuals, suggesting that sequence differences between genotypes may be driven by differing gene function. PMID- 15543587 TI - Effect of different immunisation schedules on the excretion and reversion of oral poliovaccine strains. AB - Excretion of live oral poliovaccine and molecular markers of increased virulence in the viruses isolated were examined in children who were either previously immunised with inactivated or live vaccine or were unimmunised. There appeared to be some effect of previous immunisation with either live or killed vaccine at the second dose of live vaccine. Where an effect was seen it took the form of reduced rates of excretion, shorter time periods of excretion and more rapid and complete reversion of the excreted virus. The data are consistent with the view that poliovirus is able to escape the immune pressure in the gut to some extent by improving its general fitness rather than direct evasion of immunity. PMID- 15543588 TI - Clinical and molecular characterization of chronic hepatitis B in Albania: a country that is still highly endemic for HBV infection. AB - Albania is a Mediterranean country, still with a high endemicity level of hepatitis B virus (HBV) infection. The chronic hepatitis B profile was characterized in this geographical area and used as a model to investigate the impact of endemicity level on the prevalence of the two major forms of chronic hepatitis B (HBeAg-positive and HBeAg-negative chronic hepatitis B). A cross sectional study was conducted among 62 chronic hepatitis B patients consecutively admitted to the most important tertiary health care center for the diagnosis and treatment of liver disease in Albania. HBV-DNA was measured with an in-house PCR with a sensitivity of 10(4) copies/ml which uses primers encompassing the pre core/core region. PCR products were subjected to sequencing and oligohybridization assay. Of the 62 patients, 75.8% had HBeAg-negative chronic hepatitis B. Genotype D was found in all 39 patients with detectable HBV viremia, for whom the heterogeneity of the region modulating HBeAg expression was assessed. Basic core promoter (BCP) mutations (1762/1764) were observed more often in anti-HBe-positive and older patients. In more than 90% of the HBeAg negative chronic hepatitis B patients with detectable viremia, HBV that carries the G to A pre-core mutation at nucleotide 1896 was found. Patients with HBeAg positive chronic hepatitis B were younger than HBeAg-negative chronic hepatitis B patients, and for symptomatic and asymptomatic liver-disease patients, the age of peak prevalence was at least 10 years lower for HBeAg-positive chronic hepatitis B patients. In conclusion, the virological and clinical pattern of chronic hepatitis B in Albania is similar to that observed in other Mediterranean countries; it seems to be independent of the HBV endemicity level. PMID- 15543589 TI - Impact of human metapneumovirus in childhood: comparison with respiratory syncytial virus and influenza viruses. AB - This study evaluated the overall impact of human metapneumovirus (hMPV) infection in 1,505 children and their households, and compared it with infections due to respiratory syncytial virus (RSV) and influenza viruses. Nasopharyngeal swabs were used at enrollment to collect specimens for the detection of hMPV, RSV, and influenza virus RNA by reverse-transcriptase polymerase chain reaction (RT-PCR). hMPV was detected in 42 children (2.8%), RSV in 143 (9.5%; P < 0.0001 vs. hMPV), and influenza viruses in 230 (15.3%; P < 0.0001 vs. hMPV). Of the 42 hMPV positive samples, one was also positive for RSV and six for influenza viruses, for a co-infection rate of 16.7%. Clinically, hMPV was identified only in patients with acute respiratory infection, whereas RSV and influenza viruses were also detected in patients with different clinical manifestations. Symptoms with statistically significant different proportions at presentation were fever (more frequent in the hMPV- and influenza-positive children) and wheezing with bronchiolitis or asthma exacerbation (more frequent among hMPV- and RSV-positive cases). The households of the hMPV- and the influenza-positive children had significantly more illnesses, needed significantly more medical visits, received more antipyretics, and missed significantly more work or school days than those of the RSV-positive children. Results show that hMPV is an emerging cause of acute respiratory infection in childhood, and may have a significant clinical and socioeconomic impact on children and their families. PMID- 15543590 TI - Protection against influenza virus infection by intranasal administration of hemagglutinin vaccine with chitin microparticles as an adjuvant. AB - Chitin in the form of microparticles (chitin microparticles, CMP) has been demonstrated to be a potent stimulator of macrophages, promoting T-helper-1 (Th1) activation and cytokine response. In order to examine the mucosal adjuvant effect of CMP co-administered with influenza hemagglutinin (HA) vaccine against influenza infection, CMP were intranasally co-administered with influenza HA vaccine prepared from PR8 (H1N1) virus. Inoculation of the vaccine with CMP induced primary and secondary anti-HA IgA responses in the nasal wash and anti-HA IgG responses in the serum, which were significantly higher than those of nasal vaccination without CMP, and provided a complete protection against a homologous influenza virus challenge in the nasal infection influenza model. In addition, CMP-based immunization using A/Yamagata (H1N1) and A/Guizhou (H3N2) induced PR8 HA-reactive IgA in the nasal washes and specific-IgG in the serum. The immunization with A/Yamagata and CMP resulted in complete protection against a PR8 (H1N1) challenge in A/Yamagata (H1N1)-vaccinated mice, while that with A/Guizhou (H3N2) and CMP exhibited a 100-fold reduction of nasal virus titer, demonstrating the cross-protective effect of CMP and influenza vaccine. It is suggested that CMP provide a safe and effective adjuvant for nasal vaccination with inactivated influenza vaccine. PMID- 15543591 TI - Early viral kinetics and treatment outcome in combination of high-dose interferon induction vs. pegylated interferon plus ribavirin for naive patients infected with hepatitis C virus of genotype 1b and high viral load. AB - An investigation was carried out to determine whether early viral monitoring could predict efficiently the virological response to combination therapy of two different regimens in treatment-naive chronic hepatitis C patients infected with genotype 1b with high baseline viral load. Patients were randomly assigned to receive interferon (IFN) alpha-2b induction (6 MU daily for 2 weeks) followed by 6 MU thrice weekly for 46 weeks (IFN/R group; n = 20), or pegylated IFN alpha-2b (1.5 microg/kg) weekly for 48 weeks (PEG/R group; n = 28), in combination with ribavirin (600-1,000 mg daily). Serum HCV RNA was quantitated at 0, 6, 12, 24, and 48 hr post-dose, weekly during the first 4 weeks, and thereafter viral kinetics were assessed every 4 weeks. The sustained virological response rates in the IFN/R and PEG/R groups were 40% (8/20) and 43% (12/28), respectively. The non virological response rates were 40% (8/20) and 39% (11/28), respectively. The cumulative virological response rates were similar in both groups. Multivariate analyses identified no independent baseline variables linked to sustained virological or non-virological response. Early log viral load changes from baseline in both groups were significantly greater at all time-points after 24 hr in virological response patients than in non-virological response patients (P < 0.001 for all). On the receiver operating characteristics curves for prediction of non-virological response, the area under the curves (0.951-1.000), sensitivity (90%-100%), and negative predictive value (96%- 100%) were similar at any time points after 24 hr. For prediction of sustained virological response, sensitivity of 80% with 86% negative predictive value was observed for negative HCV RNA at week 12, with the highest area under the curves value of 0.919. The results suggest that early monitoring of viral kinetics is a useful measure to predict virological response, and might facilitate development of rational and effective therapeutic strategies. PMID- 15543592 TI - Association between enterovirus endomyocardial infection and late severe cardiac events in some adult patients receiving heart transplants. AB - Enteroviruses and other cardiotropic viruses have been associated with the development of late severe adverse cardiac events in infants receiving heart transplants. However, the source and the chronology of cardiac allograft infection by an enterovirus in patients receiving heart transplants remain unknown. Using RT-PCR and immunohistochemistry assays, endomyocardial tissue samples of 30 adult patients were tested to detect the presence of specific enterovirus 5' non-coding (5'NC) sequences and of VP1 capsid protein, and this at the time of cardiac transplantation and at the 12-month biopsy for graft rejection control. Moreover, the endomyocardial detection of genomic sequences of enteroviruses, Epstein-Barr virus, herpes simplex virus, cytomegalovirus (CMV), varicella-zoster virus, adenoviruses, and parvovirus B19 was carried out by RT PCR and polymerase chain reaction (PCR) assays at the time of late severe cardiac events. Enterovirus RNA and VP1 antigen were both detected in 4 (13%) of 30 patients at the time of the 12-month biopsy for graft rejection control, whereas no enterovirus component was detected in the explanted and implanted heart tissues taken from these 4 patients at the time of transplantation. At the time when severe cardiac events were developed, within 3 months after the positive enterovirus cardiac detection, these four patients demonstrated the presence of endomyocardial enterovirus RNA sequences whereas they were tested negative for the endomyocardial detection of genomic sequences from DNA viruses (except for CMV in two cases), and for a significant level of pp65 CMV antigenemia. Taken together, these findings indicate that enteroviruses could be acquired as a new endomyocardial infection within 12 months after transplantation in adults receiving heart transplants, and suggest that this infection might be an etiological cause for unexplained late severe adverse cardiac events in the heart transplantated adults. PMID- 15543593 TI - Rho-GTPases: new members, new pathways. AB - Proteins comprising the Rho family of GTPases mediate reorganization of the actin cytoskeleton as well as transcription of genes. Recent findings from genome sequencing efforts, genetic screens, and signal transduction research have revealed that the Rho family contains several new, hitherto unrecognized members. In this review, we focus on these newly discovered Rho-GTPases and discuss their role in signaling to the cytoskeleton and the nucleus. PMID- 15543594 TI - Nucleotide correlation between HAV isolates from human patients and environmental samples. AB - Hepatitis A disease is endemic in Albania even though records from the Medical Authority show outbreaks involving only limited numbers of people. In the city of Lac, 200 people became ill following an outbreak of hepatitis A started on November 2002. The age distribution of the cases shows a peak in the age group 5 9 (43.5%) followed by the age group 10-14 (28%) and the age group 1-4 (18%). No cases were recorded in the age group 0-1. Drinking water and sewage samples were collected, using electropositive cartridges, during the outbreak. Rotavirus was identified by RT-PCR in two out of five drinking water samples; however, all the samples tested negative for HAV and astrovirus. Rotavirus was also identified in three out of five sewage samples from which four were HAV positive. HAV-RNA was identified in 28 (62.2%) out of 45 IgM anti-HAV positive sera collected during the outbreak. Genotype IA was expressed from all the amplified samples. Sequence analysis of the overlapping VP1-2A region shows 97-99% homology with three Italian strains IT-SCH-00, IT-ZAM-01, and IT-CAP-00 and one Spanish strain (Sa 30/06/95), whereas the phylogenetic tree built from the 168 bp sequence of the VP1-2A shows four clusters, one including 24 out of 28 RT-PCR positive sera. Considering the deduced amino acid sequence, only one substitution was identified and reported previously for genotype IB. PMID- 15543595 TI - Extended release of a novel antidepressant, venlafaxine, based on anionic polyamidoamine dendrimers and poly(ethylene glycol)-containing semi interpenetrating networks. AB - The multiple daily administration of venlafaxine, a novel third-generation antidepressant, was reduced based on polyamidoamine and polyethylene glycol (PEG) containing semi-interpenetrating network (IPN), respectively. Venlafaxine was covalently linked to water-soluble G2.5 anionic polyamidoamine dendrimer via a hydrolyzable ester bond. Semi-IPN hydrogels were prepared by crosslinking acrylamide in the presence of PEG, and venlafaxine with predetermined amounts was loaded in situ. Dendrimer-venlafaxine conjugate and semi-IPNs were characterized by proton nuclear magnetic resonance and Fourier transform infrared, respectively. The effect of PEG concentration and molecular weight was studied and discussed for an optimal controlled release. PMID- 15543596 TI - High-resolution MRI reflects myeloarchitecture and cytoarchitecture of human cerebral cortex. AB - Maps of cytoarchitectonically defined cortical areas have proven to be a valuable tool for anatomic localization of activated brain regions revealed by functional imaging studies. However, architectonic data require observations in a sample of postmortem brains. They can only be used reliably for comparison with functional data as probabilistic maps after spatial normalization to a common reference space. The complete architectonic analysis of an individual living brain has not been achievable to date, because the relationship remains unclear between laminar gray value changes of cerebral cortex in magnetic resonance (MR) images and those of cyto- and myeloarchitectonic histologic sections. We examined intensity profiles through the cortex in five imaging modalities: in vivo T1 and postmortem T2 MRI, one cell body stain, and two myelin stains. After visualizing the dissimilarities in the shapes of these profiles using a canonical analysis, differences between the profiles from the different image modalities were compared quantitatively. Subsequently, the profiles extracted from the in vivo T1 weighted images were estimated from profiles extracted from cyto- and myeloarchitectonic sections using linear combinations. We could verify statistically the mixed nature of the cortical T1 signal obtained in vivo: The MR intensity profiles were significantly more similar to myeloarchitectonic than to cytoarchitectonic profiles, but a weighted sum of both fitted the T1 profiles best. PMID- 15543597 TI - A recurrent duodenal gastrointestinal stromal tumor with a frameshift mutation resulting in a stop codon in KIT exon 13. AB - Gastrointestinal stromal tumors (GISTs) are a specific and rare subset of human gastrointestinal tract tumors. Most GISTs show gain-of-function mutations of KIT, mainly in exon 11, that always maintain the reading frame. We report on data from a 43-year-old Japanese man with recurrent duodenal GIST and a frameshift mutation in KIT exon 13 together with an in-frame deletion in KIT exon 11 detected by genomic DNA sequencing. Deletion of 48 base pairs of KIT exon 11, which preserved the reading frame, was identified in both primary and recurrent tumors, whereas deletion of one nucleotide of codon 642 of KIT exon 13, which changed the reading frame and induced a novel stop codon at amino acid 644, was found only in the recurrent tumor. The predicted protein resulting from the latter would lack part of the kinase domain. To the best of our knowledge, this is the first documentation of a GIST with a frameshift mutation of KIT. PMID- 15543598 TI - Genetic reconstruction of the aerobic central metabolism in Escherichia coli for the absolute aerobic production of succinate. AB - Most reported efforts to enhance production of the industrially valuable specialty chemical succinate have been done under anaerobic conditions, where E. coli undergoes mixed-acid fermentation. These efforts have often been hampered by the limitations of NADH availability, poor cell growth, and slow production. An aerobic succinate production system was strategically designed that allows E. coli to produce and accumulate succinate efficiently and substantially as a product under absolute aerobic conditions. Mutations in the tricarboxylic acid cycle (sdhAB, icd, iclR) and acetate pathways (poxB, ackA-pta) of E. coli were created to construct the glyoxylate cycle for aerobic succinate production. Experiments in flask studies showed that 14.28 mM of succinate could be produced aerobically with a yield of 0.344 mole/mole using 55 mM glucose. In aerobic batch reactor studies, succinate production rate was faster, reaching 0.5 mole/mole in 24 h with a concentration of 22.12 mM; further cultivation showed that succinate production reached 43 mM with a yield of 0.7. There was also substantial pyruvate and TCA cycle C(6) intermediate accumulation in the mutant. The results suggest that more metabolic engineering improvements can be made to this system to make aerobic succinate production more efficient. Nevertheless, this aerobic succinate production system provides the first platform for enhancing succinate production aerobically in E. coli based on the creation of a new aerobic central metabolic network. PMID- 15543600 TI - Cytogenetic features of de novo CD5-positive diffuse large B-cell lymphoma: chromosome aberrations affecting 8p21 and 11q13 constitute major subgroups with different overall survival. AB - De novo CD5-positive diffuse large B-cell lymphoma (CD5(+)DLBCL) is regarded as a different clinicopathological entity from CD5-negative DLBCL (CD5(-)DLBCL) and mantle cell lymphoma (MCL). Because only a few published cytogenetic studies of de novo CD5(+)DLBCL are available, we investigated chromosomal changes in 23 Japanese patients who had de novo CD5(+)DLBCL. A characteristic of cytogenetic abnormalities in de novo CD5(+)DLBCL was a high incidence of chromosomal aberrations affecting 8p21 and 11q13. Major chromosomal breakpoints were concentrated at 8p21, 11q13, and 3q27. Patients with 8p21 aberrations showed aggressive clinical features, including advanced stage of disease, elevated serum LDH level, poor performance status, and an inferior survival curve compared with patients who had 11q13 changes (P = .043). Chromosomal abnormalities of both 8p21 and 11q13 were not observed in the same patient, and each abnormality showed different chromosomal gains and losses. These results indicate that de novo CD5(+)DLBCL may occur in previously unidentified subgroups that differ in their chromosomal abnormalities. The conflicting results of previous studies on prognosis may thus be explained in part by the differences in chromosomal changes. PMID- 15543601 TI - Pigment pattern formation in zebrafish during late larval stages: a model based on local interactions. AB - We present a model to explain pigment cell pattern formation in the late larval stages of zebrafish, Danio rerio, and its mutants, based on new time sequence experimental data. The model stresses the importance of differential intercellular adhesion and the mechanisms of stem cell regulation to explain pigment pattern formation in zebrafish. All interactions included in the model are local in contrast to the previous models for pattern formation, which are based on long range interactions. The hypotheses of the model are tested through a mathematical (cellular automaton) model. Simulations show that differential cellular adhesion together with an appropriate mechanism of stem cell regulation are able to reproduce the main characteristics of pattern formation in the wild type zebrafish and mutants. The results obtained with the simulations are compared with experimental data and specific experimental tests to the predictions of the model are provided. PMID- 15543602 TI - Translocation t(X;11)(q13;q23) in B-cell chronic lymphocytic leukemia disrupts two novel genes. AB - Deletion of chromosome region 11q22-q23 defines a subgroup of patients with B cell chronic lymphocytic leukemia (B-CLL) characterized by poor survival. Although the tumor-suppressor gene ATM in the consensus deletion region was found to be biallelically inactivated in about one third of B-CLL cases, in the majority of those who have this deletion, inactivation of the remaining ATM allele was not observed. To identify a second disease-associated gene, we investigated two B-CLL cases with translocation breakpoints in the critical 11q23 deletion region. In one case, a t(X;11)(q13;q23) was cloned and two novel genes were isolated. The breakpoint on 11q23 affected the ARHGAP20 gene, which encodes a protein predicted to be involved in the regulation of Rho family GTPases. The breakpoint on Xq13 occurred in BRWD3, which codes for a putative novel transcription factor. The rearrangement of ARHGAP20 and BRWD3 did not result in fusion transcripts, but it disrupted both genes. Mutation analysis of 28 B-CLL samples with monoallelic deletions and two B-CLL samples with 11q23 translocations detected no deleterious mutation in the remaining copy of ARHGAP20. Quantitative expression analysis in 22 B-CLLs revealed significant up regulation of ARHGAP20 in CLL B cells, whereas BRWD3 was slightly down-regulated. Thus, deregulation of ARHGAP20 by altered gene expression or by gene disruption (but not point mutation) might be a general molecular mechanism of B-CLL leukemogenesis. PMID- 15543603 TI - Proliferation, activity, and osteogenic differentiation of bone marrow stromal cells cultured on calcium titanium phosphate microspheres. AB - In this study, the behavior of bone marrow stromal cells cultured on calcium titanium phosphate (CTP) microspheres was analyzed. Cell adhesion and proliferation were estimated by the neutral red assay and by total DNA quantification. Morphology and deposition of extracellular matrix were assessed by confocal laser scanning microscopy and/or scanning electron microscopy. The expression of the osteoblastic phenotype was evaluated by monitoring alkaline phosphatase activity and osteocalcin secretion. Results revealed that cells were able to attach and spread on the surface of CTP microspheres, and gradually grow into nearly confluent monolayers. Moreover, cells were able to bridge adjacent microspheres forming microsphere-cell clusters. Cells produced an abundant amount of fibrillar extracellular matrix that covered the substrate surface. Alkaline phosphatase activity peaked around days 7-14 and then decreased until day 21. Cells secreted osteocalcin, with higher levels being detected at day 14 than at day 21. Taken together, these results suggest that CTP microspheres are appropriate scaffolds for the growth and differentiation of cells along the osteoblastic lineage. PMID- 15543604 TI - Specification of the enveloping layer and lack of autoneuralization in zebrafish embryonic explants. AB - We have analyzed the roles of cell contact during determination of the outermost enveloping layer (EVL) and deeper neurectoderm in zebrafish embryos. Outer cells, but not deeper cells, are specified to express the EVL-specific marker, cyt1 by late blastula. EVL specification requires cell contact or close cell proximity, because cyt1 is not expressed after explant dissociation. The EVL may be homologous to the Xenopus epithelial layer, including the ventral larval epidermis. While Xenopus epidermal cytokeratin gene expression is activated by bone morphogenetic protein (BMP) signaling, zebrafish cyt1 is not responsive to BMPs. Zebrafish early gastrula ectodermal explants are specified to express the neural markers opl (zic1) and otx2, and this expression is prevented by BMP4. Dissociation of zebrafish explants prevents otx2 and opl expression, suggesting that neural specification in zebrafish requires cell contact or close cell proximity. This finding is in contrast to the case in Xenopus, where ectodermal dissociation leads to activation of neural gene expression, or autoneuralization. Our data suggest that distinct mechanisms direct development of homologous lineages in different vertebrates. PMID- 15543605 TI - Fluorinated polymers: liquid crystalline properties and applications in lithography. AB - Fluorinated polymers form an interesting class of materials with a wealth of unique properties including self-assembly, remarkably low surface energies, low absorbance to 157 nm UV light, and solubility in supercritical carbon dioxide. As a result many fluorinated polymers are of use in advanced technology applications. We review some of our work on the synthesis and characterization of block copolymers with fluorinated side chains, with special emphasis on surfaces formed using these polymers. The use of fluorinated polymers as photoresists for 157 nm lithography, with the possibility for processing in environmentally friendly supercritical carbon dioxide is also discussed. PMID- 15543606 TI - PDGFR-alpha signaling is critical for tooth cusp and palate morphogenesis. AB - Platelet-derived growth factor receptor alpha (PDGFR-alpha) and PDGF ligands are key regulators for embryonic development. Although Pdgfralpha is spatially expressed in the cranial neural crest (CNC)-derived odontogenic mesenchyme, mice deficient for Pdgfralpha are embryonic lethal, making it impossible to investigate the functional significance of PDGF signaling in regulating the fate of CNC cells during tooth morphogenesis. Taking advantage of the kidney capsule assay, we investigated the biological function of PDGF signaling in regulating tooth morphogenesis. Pdgfralpha and Pdgfa are specifically and consistently expressed in the CNC-derived odontogenic mesenchyme and the dental epithelium, respectively, throughout all stages of tooth development, suggesting a paracrine function of PDGF signaling in regulating tooth morphogenesis. Highly concentrated expression patterns of Pdgfralpha and Pdgfa are associated with the developing dental cusp, suggesting possible functional importance of PDGF signaling in regulating cusp formation. Loss of the Pdgfralpha gene does not affect proper odontoblasts proliferation and differentiation in the CNC-derived odontogenic mesenchyme but perturbs the formation of extracellular matrix and the organization of odontoblast cells at the forming cusp area, resulting in dental cusp growth defect. Pdgfralpha-/- mice have complete cleft palate. We show that the cleft palate in Pdgfralpha mutant mice results from an extracellular matrix defect within the CNC-derived palatal mesenchyme. The midline epithelium of the mutant palatal shelf remains functionally competent to mediate palatal fusion once the palatal shelves are placed in close contact in vitro. Collectively, our data suggests that PDGFRalpha and PDGFA are critical regulators for the continued epithelial-mesenchymal interaction during tooth and palate morphogenesis. Disruption of PDGFRalpha signaling disturbs the growth of dental cusp and interferes with the critical extension of palatal shelf during craniofacial development. PMID- 15543607 TI - Homochirality and life. AB - Before the emergence of life, left-handed amino acids (L-enantiomers) were selected and right-handed amino acids (D-enantiomers) were eliminated on the primal earth. Nevertheless, with the progress of analytical methods, D-amino acids have recently been found in higher order living organisms in the form of free amino acids, peptides, and proteins. Free D-amino acids have numerous physiological functions. D-amino acids containing animal peptides are well known as opioid peptides. D-amino acids in protein are related to aging. In this review, we describe the D-amino acids that are present and function as D-amino acid biosystems in our bodies. PMID- 15543608 TI - Unimolecular rectifiers and prospects for other unimolecular electronic devices. AB - We briefly review the progress towards useful one-molecule electronic devices, toward the ultimate reduction in integrated circuit sizes, and then describe five unimolecular rectifiers, or one-way conductors of electrical current: gamma hexadecylquinolinium tricyanoquinodimethanide, an acetyl sulfoxide derivative of this compound, 2,6-di[dibutylamino-phenylvinyl]-1-butylpyridinium iodide, dimethylanilino-aza[C60]fullerene, and a new fullerene derivative. PMID- 15543609 TI - Ligand-ligand redox interaction through some metal-cluster units. AB - The ligand-ligand redox interaction separated by di-, tri-, and hexanuclear cluster units is discussed in terms of the splitting of the redox waves (DeltaE(L)) and the comproportionation constants (Kcom(L)) of the ligand-based mixed valence state. Although two redox-active monodentate ligands in the mononuclear ruthenium(II) complexes (either cis or trans configurations) do not show appreciable splitting in their reduction waves, interestingly those separated by giant triruthenium and hexarhenium cluster units clearly show splitting. The molecular orbital considerations in the literature suggest that these units possess some pi-type molecular orbitals composed of metal dpi and possibly ligand ppi orbitals. Absence of the redox interactions in oxo-bridged diruthenium(II) and oxo-centered trirhodium(III) complexes where such pi molecular orbitals (including an antibonding one) are fully occupied, indicates the importance of empty pi* orbitals for the ligand redox communication. PMID- 15543610 TI - Butanol fermentation research: upstream and downstream manipulations. AB - An overview of advances in acetone-butanol fermentation research is presented with specific reference to the history of acetone-butanol fermentation, genetic manipulation of the butanol-producing Clostridium beijerinckii NCIMB 8052, as well as upstream and downstream processing. Specific reference is made to the development of the hyperamylolytic, hyper-"butanolagenic" C. beijerinckii BA101 strain. Amylolytic enzyme production by C. beijerinckii BA101 was 1.8- and 2.5 fold greater than that of the C. beijerinckii NCIMB 8052 strain grown in starch and glucose, respectively. We confirmed the presence of a phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) associated with cell extracts of C. beijerinckii BA101 by glucose phosphorylation by PEP and ATP-dependent glucose phosphorylation. It was found that C. beijerinckii BA101 was defective in PTS activity and that it compensates for this defect with enhanced glucokinase activity, resulting in an ability to transport and utilize glucose during the solventogenic stage. The principal problem associated with acetone-butanol fermentation by C. beijerinckii or C. acetobutylicum is butanol toxicity/inhibition to the culture. To solve this problem, we have attempted various alternative in situ/online techniques of butanol removal including membrane-based systems such as pervaporation, liquid-liquid extraction, and gas stripping. We found that gas stripping and pervaporation appear to be the most promising of the in situ acetone-butanol fermentation and recovery techniques but, in terms of cost-effective industrial applications, gas stripping appears to be the most promising. PMID- 15543611 TI - Frequent genetic and biochemical alterations of the PI 3-K/AKT/PTEN pathway in head and neck squamous cell carcinoma. AB - We investigated the status of the PI 3-kinase/AKT/PTEN signaling pathway in a series of 117 head and neck squamous cell carcinomas (HNSCC) in a search for molecular alterations in genes/proteins with potential prognostic value. For this purpose, PIK3CA and AKT2 gene amplification was assessed by multiplex and Quantitative Real-Time PCR. Protein expression of AKT, p-AKT, p110alpha and PTEN was determined by Western blot. PTEN allelic loss was evaluated by microsatellite analysis. PTEN-exon 5 was screened for point mutations by PCR-SSCP. Homozygous deletions were determined by multiplex PCR. PIK3CA gene was amplified in 43/117 (37%) fresh tumor samples, a frequency that did not differ from that found in archival premalignant tissues: 15/38 (39%); 12/40 (30%) fresh tumors harbored AKT2 gene amplification. AKT was found activated in 6/36 (17%) fresh tumor samples, when compared to their normal tissue counterparts. Of these 6 cases, 1 showed p110alpha overexpression and 5 displayed PTEN protein downregulation. Neither allelic loss (found in 11/77 informative cases) nor point mutations or homozygous deletions accounted for the reduced PTEN protein expression observed in our tumor series. The histologically normal mucosa of 4 patients displayed some of the molecular alterations analyzed. Dysregulation of the PI 3-K/AKT/PTEN pathway might contribute to early HNSCC tumorigenesis and might constitute a potential clinical target. Overall, 17/36 (47%) cases showed at least 1 of the molecular alterations studied here, which makes the PI 3-kinase-initiated signaling pathway one of the most frequently altered in HNSCC. PMID- 15543612 TI - Expression and prognostic value of lactoferrin mRNA isoforms in human breast cancer. AB - We investigated the expression levels of human lactoferrin (Lf), a steroid hormone-inducible gene product the expression of which is often altered during oncogenesis, and of Delta-lactoferrin (DeltaLf), its alternative isoform, which has been shown to be absent from tumor cell lines in commonly used human breast epithelial cell lines, using semiquantitative RT-PCR. Both mRNAs were detected but with levels of expression lower than those found in normal breast epithelial cells. This downregulation was much more visible for DeltaLf since its expression was either significantly diminished (BT-20, MCF-7 cell lines) or practically absent (MDA-MB-231, T-47D, HBL 100 cell lines). In order to determine whether Lf gene products are useful prognosic tools, we further analyzed their expression levels in 99 primary breast cancer biopsies. DeltaLf transcripts were found in all of the samples, whereas Lf transcripts were found in 88% of them. Lf and DeltaLf expression levels were positively correlated (p = 0.003). Lf expression was related to tumor type with a higher recovery in lobular-type tumors (p = 0.04). DeltaLf expression was related to the histoprognostic grading (p = 0.02). In univariate analyses, DeltaLf and Lf expressions were prognosis parameters, high concentrations being associated with a longer overall survival. PMID- 15543613 TI - Self-reported health and use of health care services in long-term cancer survivors. AB - Owing to an increasing number of long-term cancer survivors, the use of health care services and somatic health problems were compared between cancer survivors and a noncancer population. Data from the Nord-Trondelag Health Survey 2 (HUNT 2, 1995-1997) was merged with the Cancer Registry of Norway. Six cancer subgroups were constructed with diagnosis 5 years prior HUNT 2: testicular cancer (n= 59), colorectal cancer (n= 175), prostate cancer (n= 87), breast cancer (n= 258), gynaecological cancer (n= 153) and lymphoma/leukaemia (n= 83). For each cancer survivor 3 matched noncancer controls were selected from the HUNT 2 survey. The prevalence of common health problems, use of health care services and unfavourably life style parameters were compared between the 2 groups. Cancer survivors used health care services and received social welfare benefits more often than the controls. There was an increased risk of perceiving poor health after a history of cancer. Common health problems and/or unfavourable life style parameters could not explain poor health or the increased use of health care services among cancer survivors. Further studies are needed to investigate the reasons for increased use of health care services and perceived poor health in cancer survivors. PMID- 15543614 TI - Serenoa repens (Permixon) inhibits the 5alpha-reductase activity of human prostate cancer cell lines without interfering with PSA expression. AB - The phytotherapeutic agent Serenoa repens is an effective dual inhibitor of 5alpha-reductase isoenzyme activity in the prostate. Unlike other 5alpha reductase inhibitors, Serenoa repens induces its effects without interfering with the cellular capacity to secrete PSA. Here, we focussed on the possible pathways that might differentiate the action of Permixon from that of synthetic 5alpha reductase inhibitors. We demonstrate that Serenoa repens, unlike other 5alpha reductase inhibitors, does not inhibit binding between activated AR and the steroid receptor-binding consensus in the promoter region of the PSA gene. This was shown by a combination of techniques: assessment of the effect of Permixon on androgen action in the LNCaP prostate cancer cell line revealed no suppression of AR and maintenance of PSA protein expression at control levels. This was consistent with reporter gene experiments showing that Permixon failed to interfere with AR-mediated transcriptional activation of PSA and that both testosterone and DHT were equally effective at maintaining this activity. Our results demonstrate that despite Serenoa repens effective inhibition of 5alpha reductase activity in the prostate, it did not suppress PSA secretion. Therefore, we confirm the therapeutic advantage of Serenoa repens over other 5alpha reductase inhibitors as treatment with the phytotherapeutic agent will permit the continuous use of PSA measurements as a useful biomarker for prostate cancer screening and for evaluating tumour progression. PMID- 15543615 TI - Real-time quantitative PCR determination of urokinase-type plasminogen activator receptor (uPAR) expression of isolated micrometastatic cells from bone marrow of breast cancer patients. AB - Disseminated tumor cells (DTC) in bone marrow are independently related to poor outcome in patients with breast cancer. Phenotypic characterization of DTC may be useful to improve evaluation of the metastasizing potential of DTC and also to more accurately target aggressive tumor cells. DTC were screened in bone marrow aspirates from breast cancer patients by immunocytochemistry with an anticytokeratin (anti-CK) antibody (A45B/B3). Because the cell permeabilization and fixation required for intracellular CK staining is deleterious for mRNA, we used microaspiration to isolate single tumor cells stained with a monoclonal antibody directed against a membrane epitope, epithelial cell adhesion molecule (EpCAM), in CK-positive cases. Urokinase-type plasminogen activator receptor (uPAR) was quantified by real-time quantitative RT-PCR. The SKBR3 human breast cancer cell line was used to calibrate RT-PCR. A linear relationship was observed between the cycle threshold (Ct) of uPAR and 18S gene expression and SKBR3 cells spiked (1, 3, 7, 10 and 20) in control patient bone marrow. EpCAM-positive cells were aspirated in 21 out of 25 bone marrow specimens from breast cancer patients with CK-positive cells and uPAR mRNA expression was determined in 16 cases. A high level of uPAR mRNA in DTC was detected in 8 out of 16 patients (50%) and was associated with a more aggressive primary tumor phenotype (estrogen receptor [ER] negative, progesterone receptor [PR]-negative or HER2-positive) (p = 0.01). We demonstrated that real-time quantitative RT-PCR was reliably adapted to phenotype analysis of isolated micrometastatic cells. A larger study would be useful to confirm the importance of uPAR to define higher risk subgroups of breast cancer patients with micrometastatic disease. PMID- 15543616 TI - Differential expression of 37 selected genes in hormone-refractory prostate cancer using quantitative taqman real-time RT-PCR. AB - Progression of prostate cancer to androgen independence remains the primary obstacle to improved survival. The development of more effective treatments depends on our understanding of the molecular events associated with the hormone refractory stage. We quantified, among 90 screened genes, the expression of 37 target genes, using real-time quantitative RT-PCR. Gene expression was studied in 13 samples of HPRC compared to 33 clinically localised cancers and normal prostate tissue. We identify 19 genes with significant differential expression in HRPC compared to localised prostate cancer. Genes with decreased expression included receptors for growth factors, MMR genes and the serine protease hepsin. Analysis of increased gene expression confirmed the importance of AR upregulation and highlighted genes not previously linked to HRPC, including enzymes involved in steroid synthesis and the antiapoptotic factor survivin. Progression of prostate cancer to the hormone-refractory state is associated with differential gene expression, which may prove useful for both understanding disease progression and the development of new therapeutic approaches. PMID- 15543617 TI - Characterization of oncogene dysregulation in multiple myeloma by combined FISH and DNA microarray analyses. AB - Chromosomal translocations involving the immunoglobulin heavy chain (IGH) locus and various partner loci frequently are associated with multiple myeloma (MM). We investigated the expression profiles of the FGFR3/MMSET, CCND1, CCND3, MAF, and MAFB genes, which are involved in t(4;14)(p16.3;q32), t(11;14)(q13;q32), t(6;14)(p21;q32), t(14;16)(q32;q23), and t(14;20)(q32;q12), respectively, in purified plasma cell populations from 39 MMs and six plasma cell leukemias (PCL) by DNA microarray analysis and compared the results with the presence of translocations as assessed by dual-color FISH or RT-PCR. A t(4;14) was found in 6 MMs, t(11;14) in 9 MMs and 1 PCL, t(6;14) in 1 MM, t(14;16) in 2 MMs and 1 PCL, and t(14;20) in 1 PCL. In all cases, the translocations were associated with the spiked expression of target genes. Furthermore, gene expression profiling enabled the identification of putative translocations causing dysregulation of CCND1 (1 MM and 1 PCL) and MAFB (1 MM and 1 PCL) without any apparent involvement of immunoglobulin loci. Notably, all of the translocations were mutually exclusive. Markedly increased MMSET expression was found in 1 MM showing associated FGFR3 and MMSET signals on an unidentified chromosome. Our data suggest the importance of using combined molecular cytogenetic and gene expression approaches to detect genetic aberrations in MM. PMID- 15543618 TI - ERBB2 and MYC alterations in BRCA1- and BRCA2-associated cancers. PMID- 15543619 TI - Lymphoid gene expression as a predictor of risk of secondary brain tumors. AB - Gene expression profiles are tissue-specific but may also reflect germ-line driven expression patterns across tissue types. Previously, using a targeted pharmacologic approach, we identified germ-line polymorphisms in a single gene (thiopurine methyltransferase) associated with the risk of irradiation- and chemotherapy-induced secondary brain tumors in children with acute lymphoblastic leukemia (ALL). To identify additional candidate genetic risk factors, in identically treated patients, we compared the gene expression profiles of diagnostic ALL blasts of those who did develop irradiation-associated brain tumors (n = 9) with the profiles from those who did not (n = 33). Weighted rank regression was used to identify 33 probe sets associated with the time-dependent development of brain tumors; k-means clustering (k = 2) identified 2 groups that differed significantly in cumulative incidence of brain tumors (P = 0.012). Permutation analysis was used to estimate the probability (P = 0.18) of obtaining 2 such clusters by chance. Linear discriminant analysis (time-independent categorization of outcome) was used to identify 70 probe sets whose expression differentiated between the 2 groups of patients. Permutation analyses (n = 1,000) was used to estimate the probability of selecting these probe sets by chance (P = 0.055). Five probe sets were in common between the time-independent and time dependent methods. The distinguishing genes are involved in neural growth (FGFR1) and in nuclear trafficking (HNRPL, KPNB1). These data suggest that gene expression profiling from accessible tissues may identify targets involved in therapy-related malignancies in unrelated tissues. PMID- 15543620 TI - CDK4 is a probable target gene in a novel amplicon at 12q13.3-q14.1 in lung cancer. AB - Several chromosomal regions are recurrently amplified or deleted in lung tumors, but little is known about the underlying genes, which could be important mediators in tumor formation or progression. In lung cancer, the RB1-CCND1-CDKN2A pathway, involved in the G1-S transition, is damaged in nearly all tumors. In the present study, we localized a novel amplicon in lung tumors to a fragment of less than 0.5 Mb at 12q13.3-q14.1 by using comparative genomic hybridization (CGH) on cDNA microarrays. This approach enabled us to identify 10-15 genes with the most consistent amplifications. Semiquantitative RT-PCR analyses of 13 genes in this region showed that four of them (CDK4, CYP27B1, METTL1, and TSFM) were also highly up-regulated. Immunohistochemical (IHC) analysis of 141 tumor samples on a tissue microarray showed that CDK4 was expressed at a high level in 23% of lung tumors. Six (21.4%) of the tumors with high CDK4 expression (n = 28) were shown by fluorescence in situ hybridization (FISH) to contain the 12q13.3-q14.1 amplification. For CDK4, a positive correlation was found between gene copy number (FISH and CGH array), mRNA expression (RT-PCR), and level of protein expression (IHC). CDK4 expression did not correlate with CDKN2A methylation status. Amplification of CDK4 has been described in other tumor types, but its role in lung cancer remains to be elucidated. Although CDK4 amplification seems to be a relatively rare event (4.3%) in lung tumors, it indicates the significance of the RB1-CCND1 pathway in lung tumorigenesis. PMID- 15543621 TI - Re: Gutierrez-Enriquez S, Fernet M, Dork T, Bremer M, Lauge A, Stoppa-Lyonnet D, Moullan N, Angele S, Hall J, "Functional consequences of the ATM sequence variants for chromosomal radiosensitivity". PMID- 15543623 TI - Complex biallelic IGH rearrangements in IgM-expressing Z-138 cell line: Involvement of downstream immunoglobulin class switch recombination. AB - Chromosomal translocations involving the immunoglobulin (Ig) receptor loci usually disrupt and silence these loci. On the basis of observations in follicular lymphoma (FL) with downstream Ig heavy chain (IGH) class switch recombination (CSR), we hypothesized that downstream CSR-mediated chromosomal translocations would leave the V(D)J-Cmu transcription unit intact, thereby still allowing IgM expression from the IGH allele involved in the translocation. To test this hypothesis, we analyzed biallelic IGH translocations in the IgM expressing cell line Z-138 by interphase FISH, DNA fiber-FISH, long-distance vectorette PCR, and DNA sequencing. One IGH allele was involved in a t(11;14), showing a break in the JH region that juxtaposed the Emu enhancer and the 3' Calpha enhancers to the cyclin D1 gene. The other IGH allele contained a t(8;14) breakpoint involving the 3' end of a Sgamma region, whereas the reciprocal breakpoint at 8q24 was approximately 40 kb centromeric of MYC. Molecular analysis showed that this IGH allele harbored a normal V(D)J-Cmu complex, which is responsible for IgM expression. These data show that chromosomal breakpoints such as the t(8;14) can occur in downstream IGH constant regions and do not necessarily interfere with Ig expression. PMID- 15543624 TI - Formation of der(19)t(1;19)(q23;p13) in acute lymphoblastic leukemia. AB - The t(1;19)(q23;p13), which results in a fusion of TCF3 (previously E2A) at 19p13 with PBX1 at 1q23, is one of the most common translocations in acute lymphoblastic leukemia (ALL). It is seen either as a balanced t(1;19) or as an unbalanced der(19)t(1;19); occasional cases with coexisting t(1;19)- and der(19) positive clones also have been described. Although it generally has been assumed that the unbalanced form arises from the balanced t(1;19) through loss of the derivative chromosome 1 followed by duplication of the normal homologue, this has never been proved. At least two other mechanisms are possible for the formation of the der(19): an initial trisomy 1 followed by translocation and subsequent loss of the der(1) or a rearrangement during the G2 phase of the cell cycle, with the derivative chromosomes 1 and 19 ending up in separate daughter cells. The different alternatives may be distinguished by investigation of markers proximal to the breakpoint in 1q23 because they would be expected to lead to different allelic patterns. Thus, loss of heterozygosity as a result of the presence of uniparental disomy (UPD)-both copies of a chromosome being derived from only one parent-for chromosome 1 would be present in all der(19)-harboring cases arising via the duplication pathway and in one-third of cases arising via the trisomy pathway, but in none of the der(19) formed via the G2 pathway. In this study, we used quantitative fluorescence PCR with polymorphic microsatellite markers to investigate chromosomes 1 and 19 in two t(1;19)- and four der(19)-positive ALLs. None of the der(19) cases displayed UPD for chromosome 1, excluding that this aberration arises through the duplication pathway. Because previous findings of cases with coexisting t(1;19) and der(19) clones are difficult to explain if the translocation originated in G2, the present results suggest that an unbalanced der(19) may arise from an initial trisomy 1 followed by t(1;19) translocation and loss of the derivative chromosome 1. PMID- 15543625 TI - Hydrolytic resolution of (R,S)-naproxen 2,2,2-trifluoroethyl thioester by Carica papaya lipase in water-saturated organic solvents. AB - For the first time, the Carica papaya lipase (CPL) stored in crude papain is explored as a potential enantioselective biocatalyst for obtaining chiral acids from their racemic thioesters. Hydrolytic resolution of (R,S)-naproxen 2,2,2 trifluoroethyl thioester in water-saturated organic solvents is employed as a model system for studying the effects of temperature and solvents on lipase activity and enantioselectivity. An optimal temperature of 60 degrees C, based on the initial rate of (S)-thioester and a high enantiomeric ratio (i.e., E-value defined as the ratio of initial rates for both substrates) of >100 at 45 degrees C in isooctane, is obtained. Kinetic analysis, considering product inhibition and enzyme deactivation, is also performed, showing agreement between the experimental and best-fit conversions for (S)-thioester. A comparison of the kinetic and thermodynamic behaviors of CPL and Candida rugosa lipase (CRL) in isooctane and cyclohexane indicates that both lipases are very similar in terms of thermodynamic parameters DeltaDeltaH and DeltaDeltaS, initial rate of (S) substrate, and E-value when (R,S)-naproxen 2,2,2-trifluoroethyl thioester or ester is employed as substrate. PMID- 15543627 TI - Estimation of endogenous noradrenaline release in rat brain in vivo using [3H]RX 821002. AB - Noradrenaline plays an important role in many normal brain functions, e.g., attention, memory, and emotion. Dysfunction in the noradrenergic system is thought to lead to a number of abnormal brain conditions. The lack of suitable in vivo tracers to monitor noradrenaline release, levels, and regulation has hampered our fully understanding the roles that it plays in the brain. Presented here are data showing that the in vivo binding of the alpha2-adrenoceptor antagonist [3H]RX 821002 is sensitive to endogenous noradrenaline. Elevation of extracellular noradrenaline, using three different pharmacological challenges in rat, led to a reduction in the binding potential (BP) of [3H]RX 821002 when compared with vehicle controls. The challenges used were i.p. administration of D amphetamine, the imidazoline2 binding site-selective ligand BU224, and L deprenyl. Of the cortical regions measured, the reduction in BP reached significance in the anterior cingulate cortex for all of these pharmacological challenges. These initial observations in rat indicate that labelling of the alpha2-adrenoceptors with RX 821002 can be used to estimate changes in extracellular noradrenaline concentration in the cortex. This has the potential to enable the investigation of the role that noradrenaline plays both in the normal and abnormal brain and, if the ligand can be radiolabelled with a suitable positron-emitting isotope at high specific radioactivity, it could be an invaluable PET tracer. PMID- 15543628 TI - First visualization of adenosine A(2A) receptors in the human brain by positron emission tomography with [11C]TMSX. AB - [11C]TMSX is a new positron emission tomography (PET) radioligand that provides visualization of adenosine A(2A) receptors (A(2A)Rs) in the brain, heart and skeletal muscle. Here we report on the first visualization of the A(2A)Rs in the human brain by PET and [11C]TMSX in a male healthy volunteer, compared with the adenosine A1 receptors (A1Rs) and dopamine D2 receptors (D2Rs) which were measured by PET with [11C]MPDX and [11C]raclopride, respectively. The distribution volume (DV) of [11C]TMSX in the baseline was relatively high in the head of caudate nucleus, putamen, and thalamus and relatively low in the cortical regions. Infusion of theophylline, a nonselective A(2A)R antagonist (Ki for A(2A)Rs = 16000 nM for theophylline vs 5.9 nM for TMSX), slightly reduced the DVs in the head of caudate nucleus (8.0% reduction) and putamen (4.5% reduction), but not in the other regions having much lower levels of A(2A)Rs, demonstrating the A(2A)R-specific binding of [11C]TMSX. On the other hand, the A1Rs were widely distributed in the whole brain except for the cerebellum, while the binding potential of [11C]raclopride was predominantly high in the striatum. We concluded that [11C]TMSX is an applicable PET ligand for mapping the A(2A)Rs in the caudate nucleus and putamen in clinical studies because of no availability of other radioligands until now. The [11C]TMSX PET is of great interest for studying the pathophysiology of neurological and psychiatric disorders together with the [11C]raclopride PET for D2Rs evaluation and/or the [11C]MPDX PET for A1Rs evaluation. PMID- 15543629 TI - Correlation of non-uniform protein expression with variation in transmitter release probability. AB - The strength of synaptic transmission is highly variable between different synapses. The present study examined some factors that may contribute to this variation in the strength of neurotransmission in sympathetic varicosities of the mouse vas deferens. Transmitter release was measured using a focal macropatch electrode placed over pairs of visualised varicosities. By regulating the calcium concentration of the solutions inside the recording electrode and in the bath independently of each other, transmitter release was restricted to one or two surface varicosities at each recording site. Using this technique, transmitter release probability was shown to be highly variable, even between adjacent varicosities on single axon branches. Very little variation was observed in the calcium influx following single impulse nerve stimulation between adjacent Oregon Green BAPTA-1 loaded varicosities. However, the staining intensities of three vesicular proteins, SV2, synaptophysin, and synaptotagmin 1, showed considerable variation between adjacent varicosities on single axon branches. This variation in staining intensity may be partly explained by variation in the density of synaptic vesicles. However, double staining experiments using two vesicular antigens showed some varicosities staining for one vesicular antigen, but not for the second, suggesting that the expression of these release machinery proteins is regulated locally within the varicosities. The results of the present study strengthen suggestions that synaptic strength is at least in part, regulated by variation in the expression of vesicular proteins. PMID- 15543630 TI - Safety and efficacy of gamma-vinyl GABA (GVG) for the treatment of methamphetamine and/or cocaine addiction. AB - This study examined the safety and efficacy of gamma vinyl-GABA (GVG, vigabatrin) for the treatment of methamphetamine and/or cocaine addiction. A total of 30 subjects, who met DSM-IV criteria for methamphetamine and/or cocaine dependence, were enrolled in an open label 9-week safety study. The protocol was specifically designed to include extensive visual field monitoring as well as outcome measures of therapeutic efficacy. Patients were screened twice weekly for the presence of urinary cocaine, methamphetamine, heroin, alcohol, and marijuana. In total, 18/30 subjects completed the study and 16/18 tested negative for methamphetamine and cocaine during the last 6 weeks of the trial. GVG did not produce any visual field defects or alterations in visual acuity. Furthermore, it did not produce changes in vital signs even with continued use of methamphetamine and cocaine. Thus, under conditions that appear to be appropriate for the successful treatment of methamphetamine and/or cocaine addiction, GVG is safe. PMID- 15543631 TI - Development and validation of vertical scanning interferometry as a novel method for acquiring chondrocyte geometry. AB - Chondrocytes are sensitive to changes in shape which depend on the type of substrate, mechanical factors, or biochemical stimuli. Shape changes can cause metabolic and phenotypic alterations. Cell geometry is also important for mechanical models, determination of mechanical properties, and the study of cell attachment and spreading. In this study, a novel method called vertical scanning interferometry (VSI) was developed to allow rapid and straightforward determination of the height, diameter, surface area, volume, and curvature of single chondrocytes. The dimensions of single chondrocytes at 4 and 18 h were obtained and validated. Differences in the heights of zonal chondrocytes were found to be statistically significant. This method was also used to capture the geometry of a cell dehydrating as it was exposed to air. VSI has advantages over confocal microscopy and atomic force microscopy in terms of speed, ease of use, field of view, and precision. VSI quickly obtains and graphically represents the three-dimensional geometry of chondrocytes in a simple format. These methods could be expanded to image many types of cells on various biomaterials to assess biocompatibility and attachment and cell spreading characteristics. PMID- 15543632 TI - The influence of microtextured basal lamina analog topography on keratinocyte function and epidermal organization. AB - The rational design of future bioengineered skin substitutes requires an understanding of the mechanisms by which the three-dimensional microarchitecture of tissue scaffolds modulates keratinocyte function. Microtextured basal lamina analogs were developed to investigate the relationship between the characteristic topography at the dermal-epidermal interface of native skin and keratinocyte function. Microfabrication techniques were used to create master patterns, negative replicates, and collagen membranes with ridges and channels of length scales (e.g., grooves of 50-200 microm in depth and width) similar to the invaginations found in basal lamina at the dermal-epidermal junction of native skin. Keratinocytes were seeded on the surfaces of basal lamina analogs, and histological analyses were performed after 7 days of tissue culture at the air liquid interface. The keratinocytes formed a differentiated and stratified epidermis that conformed to the features of the microtextured membranes. Morphometric analyses of immunostained skin equivalents suggest that keratinocyte stratification and differentiation increases as channel depth increases and channel width decreases. This trend was most pronounced in channels with the highest depth-to-width ratios (i.e., 200 microm deep, 50 microm wide). It is anticipated that the findings from these studies will elucidate design parameters to enhance the performance of future bioengineered skin substitutes. PMID- 15543633 TI - Accelerated bone repair with the use of a synthetic BMP-2-derived peptide and bone-marrow stromal cells. AB - A novel synthetic peptide corresponding to BMP-2 residues 73-92 that can induce bone formation and can form a conjugate with a carrier to localize its effect has been reported previously. The synthetic peptide was bound to a BMP-2-specific receptor, and it elevated both the alkaline phosphatase activity and the osteocalcin mRNA in the murine multipotent mesenchymal cell line, C3H10T1/2. The 73-92 peptide also induced ectopic bone formation when conjugated to a covalently crosslinked alginate gel and implanted into a rat's calf muscle. Here, it is reported that the 73-92 peptide-conjugated alginate gel particles significantly promoted the repair of rat tibial bone defects, whereas the alginate gel sponge that the peptide was conjugated with was less effective. Further acceleration and denser bone regeneration was achieved when the 73-92 peptide-conjugated alginate gel particles were coimplanted with syngeneic rat bone-marrow stromal cells. Therefore, the 73-92 peptide can induce differentiation of osteoblast precursor cells into osteoblasts, and can activate osteoblasts to promote the repair of bone defects. PMID- 15543634 TI - Verification bias-corrected estimators of the relative true and false positive rates of two binary screening tests. AB - The relative accuracy of two binary screening tests can be quantified by estimating the relative true positive rate (rTPR) and relative false positive rate (rFPR) between the two tests. Ideally all study subjects are administered both screening tests as well as a gold standard to determine disease status. In practice, however, often the gold standard is so invasive or costly that only a percentage of study subjects receive disease verification and the percentage differs depending on the results of the two screening tests. This is known as verification-biased sampling and may be by design or due to differential patient dropout or refusal to have the gold standard test administered. In this paper, maximum likelihood estimators of rTPR and rFPR and corresponding confidence intervals are developed for studies with verification-biased sampling assuming that disease status is missing at random (MAR). Simulation studies are used to show that if the MAR assumption holds, then the verification bias-corrected point estimators have little small sample bias and the confidence intervals have good coverage probabilities. Simulation studies also demonstrate that the verification bias-corrected point estimators may not be robust to violation of the MAR assumption. The proposed methods are illustrated using data from a study comparing the accuracy of Papanicolaou and human papillomavirus tests for detecting cervical cancer. PMID- 15543635 TI - SNPs, haplotypes, and model selection in a candidate gene region: the SIMPle analysis for multilocus data. AB - Modern molecular techniques make discovery of numerous single nucleotide polymorphims (SNPs) in candidate gene regions feasible. Conventional analysis relies on either independent tests with each variant or the use of haplotypes in association analysis. The first technique ignores the dependencies between SNPs. The second, though it may increase power, often introduces uncertainty by estimating haplotypes from population data. Additionally, as the number of loci expands for a haplotype, ambiguity in interpretation increases for determining the underlying genetic components driving a detected association. Here, we present a genotype-level analysis to jointly model the SNPs via a SNP interaction model with phase information (SIMPle) to capture the underlying haplotype structure. This analysis estimates both the risk associated with each variant and the importance of phase between pairwise combinations of SNPs. Thus, rather than selecting between genotype- or haplotype-level approaches, the SIMPle method frames the analysis of multilocus data in a model selection paradigm, the aim to determine which SNPs, phase terms, and linear combinations best describe the relation between genetic variation and a trait of interest. To avoid unstable estimation due to sparse data and to incorporate both the dependencies among terms and the uncertainty in model selection, we propose a Bayes model averaging procedure. This highlights key SNPs and phase terms and yields a set of best representative models. Using simulations, we demonstrate the utility of the SIMPle model to identify crucial SNPs and underlying haplotype structures across a variety of causal models and genetic architectures. PMID- 15543637 TI - Genetic epidemiology and haplotypes. PMID- 15543638 TI - Evaluating associations of haplotypes with traits. AB - Haplotypes have played a major role in the study of highly-penetrant single-gene disorders, and recent evidence that the human genome has hot-spots and cold-spots for recombination have suggested that haplotype-based methods may play a key role in the study of common complex traits. This report reviews the motivation of using haplotypes for the study of the genetic basis of human traits, ranging from biologic function, to statistical power advantages of haplotypes, to linkage disequilibrium fine-mapping. Recent developments of regression models for haplotype analyses are reviewed, offering a synthesis of current methods, as well as their limitations and areas that require further research. Regression models provide significant advantages, such as the ability to control for non-genetic covariates, the effects of the haplotypes can be modeled, step-wise selection can be used to screen for a subset of markers that explain most of the association, haplotype x environment interactions can be evaluated, and regression diagnostics are well developed. Despite these strengths, the current regression methods tend to lack the sophisticated population genetic perspectives offered by coalescent and other similar approaches. Future work that links regression methods with population genetic models may prove beneficial. PMID- 15543639 TI - Two-Stage sampling designs for gene association studies. AB - We consider two-stage case-control designs for testing associations between single nucleotide polymorphisms (SNPs) and disease, in which a subsample of subjects is used to select a panel of "tagging" SNPs that will be considered in the main study. We propose a pseudolikelihood [Pepe and Flemming, 1991: JASA 86:108-113] that combines the information from both the main study and the substudy to test the association with any polymorphism in the original set. SNP tagging [Chapman et al., 2003: Hum Hered 56:18-31] and haplotype-tagging [Stram et al., 2003a; Hum Hered 55:27-36] approaches are compared. We show that the cost efficiency of such a design for estimating the relative risk associated with the causal polymorphism can be considerably better than for a single-stage design, even if the causal polymorphism is not included in the tag-SNP set. We also consider the optimal selection of cases and controls in such designs and the relative efficiency for estimating the location of a causal variant in linkage disequilibrium mapping. Nevertheless, as the cost of high-volume genotyping plummets and haplotype tagging information from the International HapMap project [Gibbs et al., 2003; Nature 426:789-796] rapidly accumulates in public databases, such two-stage designs may soon become unnecessary. PMID- 15543640 TI - Allelic association patterns for a dense SNP map. AB - A dense set of 5,000 SNPs on a 10-Mb region of human chromosome 20 has been typed on samples of African Americans, East Asians, and United Kingdom Caucasians. There are departures from Hardy-Weinberg equilibrium beyond the level at which markers are often discarded because of possible genotyping errors. The observation that markers showing such departures are often close together on the chromosome confirms the result that Hardy-Weinberg tests at two loci are correlated to an extent that depends on the linkage disequilibrium between those two markers. Linkage disequilibrium can be described by the composite linkage disequilibrium coefficient, the parameter that determines the behavior of case control allelic tests of association. A useful preliminary investigation of datasets of this type is provided by counting the numbers of distinct multi-locus genotypes in windows of a few markers. PMID- 15543641 TI - Genotype prediction using a dense map of SNPs. AB - The International Haplotype Mapping Project (HapMap) aims to characterize the distribution and extent of linkage disequilibrium (LD) throughout the human genome, thereby facilitating genome-wide association analysis and the search for the genetic determinants of complex diseases. Implicit in the rationale behind the project is the expectation that hidden (unobserved) disease-causing variants will be in significant LD with surrounding typed markers and will thus be amenable to detection using association-based mapping approaches. In order to investigate the validity of this assumption, we examined more than 5,000 SNPs across a 10-MB region of chromosome 20 in a sample of 96 unrelated African American and 96 unrelated Caucasian individuals. We treated observed loci as surrogates for hidden SNPs by pretending that individuals' genotypes were unknown. We then attempted to predict these genotypes at the surrogate hidden SNP by using information about LD in the region and genotypes at surrounding observed loci. Our method is based on finding the most likely genotype for each individual, given all possible haplotype pairs consistent with observed genotypes for that individual at surrounding loci, and given the frequencies of those haplotypes in an independent sample. Our method performs extremely well in predicting genotypes in areas of high LD. Furthermore, in areas of low LD, our method results in substantial gains in predictive accuracy as compared to pair wise strategies. These results suggest that pair-wise tests of disease-marker association may be inferior to multipoint methods, which take advantage of the information contained within multi-locus haplotypes. PMID- 15543642 TI - Neutrophils from Brazilian patients with Graves' disease: some biochemical and functional aspects. AB - Graves' disease shows important systemic inflammatory complications and has been considered to be systemic autoimmune thyroid, skeletal muscle and connective tissue syndrome. Neutrophils participate in the pathophysiology of the two major immune and inflammatory manifestations of the disease, ophthalmopathy and myxedema, and may worsen the inflammatory picture. In this study we analysed some biochemical and functional aspects of neutrophils in Graves' disease patients to assess their participation in these processes. The results show that the complement and/or Fcgamma receptor-mediated oxygen radical production by neutrophils was increased when patient cells were compared with controls. However the percentage of cells expressing complement and IgG receptors and the per-cell fluorescence, were similar, indicating that the increased oxidative burst was not due to an abnormal expression of mediating receptors. The production of hydrogen peroxide was also increased in hyperthyroid patient neutrophils as compared to controls. Conversely, antioxidant defences (superoxide dismutase activity and reduced glutathione content) in neutrophils from patients were not significantly different from healthy controls. The liberation of potent oxidative compounds together with the absence of adequate quenching of them by antioxidant mechanisms could be responsible for greater tissue damage in inflammatory conditions, as is the case in ophthalmopathy and myxedema patients. Considering our results and those of other workers, we encourage and suggest an associated antioxidant therapy to complement the conventional anti-thyroid therapy, especially in obvious inflammatory cases and in individuals who smoke. PMID- 15543643 TI - DNA polymorphism of Pvu II site in the lipoprotein lipase gene in patients with non-insulin dependent diabetes mellitus. AB - We studied the effect of variation at the lipoprotein lipase (LPL) gene locus on the susceptibility of individuals with non-insulin dependent diabetes mellitus (NIDDM) in a population of 110 NIDDM patients and 91 controls. Our objective was to study the relationship between the LPL-Pvu II polymorphism and NIDDM and lipid metabolism. PCR-RFLP was used to determine the DNA polymorphism of the sixth intron of the LPL gene. The frequencies of the genotypes in case and control groups were 29.1 and 30.8% for P+/P+; 45.5 and 36.3% for P+/P-; 25.5 and 33% for P-/P- respectively. There was no significant difference in frequencies of genotypes between the two groups. Logistic regression analysis revealed that triacylglycerol (TAG) and apolipoprotein E levels were associated with NIDDM, whereas Pvu II genotypes were not found as independent risk factors for the disease. Overall this study demonstrates the role of the Pvu II polymorphism in the LPL gene in modulating plasma lipid/lipoprotein levels in patients with NIDDM. PMID- 15543644 TI - Comparative acute effects of leptin and insulin on gluconeogenesis and ketogenesis in perfused rat liver. AB - The acute effects of physiological levels of leptin (10 ng ml(-1)) and insulin (20 microU ml(-1)) on hepatic gluconeogenesis and ketogenesis were compared. Leptin or insulin alone decreased (p<0.05) the activation of hepatic glucose, L lactate and urea production from L-alanine. However, the hepatic glucose production was not modified if leptin was combined with insulin. These results indicated that both, i.e. leptin and insulin, could promote a non-additive reduction in the rate of catabolism of L-alanine. However, in contrast with insulin (p<0.05), leptin did not inhibit the activation of hepatic glucose production from pyruvate or glycerol. On the other hand, activation of hepatic production of acetoacetate and beta-hydroxybutyrate from octanoate was not affected by leptin or insulin. Thus, our data demonstrate that the acute effect of leptin on hepatic metabolism was partially similar to insulin (activation of glucose production from L-alanine and activation of acetoacetate or beta hydroxybutyrate production from octanoate) and partially different from insulin (activation of glucose production from pyruvate or glycerol). PMID- 15543645 TI - Tandem (quadrupole-time-of-flight) electrospray mass spectrometry of oligo(vinyl acetates) with isopropylol or (1-hydroxyethyl)-2-oxyisopropanyl end groups. AB - Poly(vinyl acetates) with either isopropylol or (1-hydroxyethyl)-2-oxyisopropanyl end groups were analysed by tandem mass spectrometry using a quadrupole-time-of flight (Q-TOF) instrument. Random scission along the polymer backbone was not observed. Instead the initial scissions were derived from the cleavage of end group species. The resultant macrocations were subject to further elimination reactions that eventually produced polyene macrocations. A smaller fraction of the initial macrocations also reacted by loss of ketene and this reaction produced macrocations containing vinyl alcohol units. PMID- 15543646 TI - Enatiomeric analysis of simendan by CE with beta-CD as chiral selector compared with CMPA-HPLC. AB - The chiral separation of simendan enantiomers using capillary electrophoresis was studied with beta-cyclodextrin (beta-CD) as chiral selector. The influences of the concentration and pH of borate buffer solution, beta-CD concentration and methanol content in the background electrolyte were investigated. These factors were compared with those in an HPLC with beta-CD as chiral mobile phase additive (CMPA-HPLC). The quantification properties of the developed CE method were examined. A baseline separation of simendan enantiomers was achieved in the background electrolyte of 20 mmol/L borate buffer (pH 11.0) containing 12 mmol/L beta-CD-methanol (50:50 in volume ratio). The CE method is comparable with CMPA HPLC in chiral resolution, although the optimal pH in CE (11.0) is much higher than that (6.0) in CMPA-HPLC. This chiral CE method is applicable to the quantitative ananlysis and enantiomeric excess value determination of L-simendan. PMID- 15543647 TI - [Medical development of invagination: pediatric surgeon's point of view. Commentary on Hofer H (1932) Chirurg 4:604-613]. PMID- 15543648 TI - [Liposuction]. PMID- 15543649 TI - [How do economic pressures, medical tasks and Christian ethics reconcile in the modern medicine?]. PMID- 15543650 TI - [DRG and maximal care hospitals. Extent and causes of underfinancing]. PMID- 15543652 TI - [Working hours law--what is valid?]. PMID- 15543651 TI - Slaves to fear. PMID- 15543653 TI - [Re: Disparity in expert opinion of suspected treatment errors, for example, in injuries of the N. accessorius on the occasion of lymph node sample excision in the neck Hansis & Kienzle, Chirurg BDC (2004) 43:M31-M34]. PMID- 15543654 TI - [Re: Disparity in expert opinion of suspected treatment errors, for example, in injuries of the N. accessorius on the occasion of lymph node sample excision in the neck. Hansis & Kienzle, Chirurg BDC (2004) 43:M31-M34]. PMID- 15543655 TI - Bluetooth might bite. PMID- 15543656 TI - HMBA induces cell death and potentiates doxorubicin toxicity in malignant mesothelioma cells. AB - PURPOSE: Malignant pleural mesothelioma(MM), a rare tumor characterized by high local invasiveness and low metastatic efficiency, is poorly responsive to current therapeutic approaches. The aim of the present study was to evaluate the cytotoxic efficacy of the hybrid polar compound hexamethylene bisacetamide(HMBA), either as a single agent or in combination with the anthracycline doxorubicin (DOX), against MM cells. METHODS: The MM cell lines MM-B1 and MM-El were treated with HMBA, DOX or with combinations of the two drugs. Cell survival and death were assessed by the MTS assay and trypan blue staining/TUNEL, respectively. The interactions between drugs were evaluated by the method of Kern et al. Western blot analysis was used to investigate the expression of Bcl-2 family proteins. RESULTS: When administered alone, HMBA dose-dependently decreased the number of viable cells and increased the death rate of MM-B1 and MM-E1 cultures. Combinations of HMBA and DOX achieved a synergistic inhibition of MM cell survival, and the simultaneous administration of HMBA counteracted the resistance induced by DOX in MM-El cells. HMBA,used at cytostatic concentrations, reduced the ratio be-tween antiapoptotic (Bcl-2, Bcl-XL) and proapoptotic(Bax) members of the Bcl-2 family of proteins, thus lowering the threshold for MM cell death commitment. CONCLUSIONS: HMBA has therapeutic potential in MM both as a single agent and through potentiation of DOX toxicity. These results support future investigations on the feasibility of intrapleural chemotherapy with this hybrid polar compound. PMID- 15543657 TI - Preventing mother to child transmission of HIV. PMID- 15543658 TI - Primary care physicians who treat blacks and whites. PMID- 15543659 TI - Primary care physicians who treat blacks and whites. PMID- 15543660 TI - High versus low PEEP in ARDS. PMID- 15543661 TI - Aldosterone revisited. PMID- 15543662 TI - Aldosterone revisited. PMID- 15543663 TI - Deep-vein thrombosis. PMID- 15543664 TI - Evolution: how do characters evolve? AB - Ricklefs claims to show that morphological evolution in birds is associated with speciation events--that is, it is punctuational--by inference from data on only species number, clade age and character variance from a range of passerine clades. He suggests that variance increases in proportion with clade age under gradual change, but in proportion to the logarithm of species number if change is punctuational. Here I show that both clade age and the logarithm of species number independently predict variance under both gradual and punctuational change, rendering Ricklefs' results uninformative about his central hypothesis. PMID- 15543665 TI - [Development of chromate allergies]. PMID- 15543666 TI - [Patients as partners. Tumor patients and their participation in medical decisions]. AB - Between March 2002 and August 2003 as part of the research project "Patients as partners -- tumour patients and their participation in medical decisions" tumour patients undergoing palliative therapy (n=272) were interviewed and asked about their level of information, their desired place to die and whether they had prepared an advance directive. Furthermore, 72 relatives of deceased patients who had been looked after by the project's palliative care team were given a similar questionnaire including questions concerning their knowledge about disease and prognosis, the actual place of death and the relevance of advance directives. According to patients and relatives, information particularly about prognosis is unsatisfactory. Of the inter-viewed patients, 75% said they wanted to die at home and 15% in a hospital. According to their relatives, 36% of the patients looked after by the palliative care team had an advance directive. The survey of the relatives showed a significant relation between the preparation of an advance directive and dying at the desired place. According to the relatives, medical and health reasons, hope for an improvement up to the very end,acute worsening of the condition and deficits in medical care were important reasons for dying in hospital against the patient'swish. In future, advance directives should be used as an aid for communication and the planning of care. Therefore, cooperation between doctors and patients based on a partnership is necessary. The required competence in communication should be improved. PMID- 15543667 TI - Novel ideas for fast muscle action potential simulations using the line source model. AB - Using a signal processing approach, we analyze the line source model for muscle action potential (AP) modeling. We show that the original model presents a tradeoff between violating the Nyquist criterion on one hand and using a discretization frequency that is unnecessarily high with respect to the bandwidth of the generated AP on the other. Here, we present an improved line source model that, compared to the original, allows a lower discretization frequency while retaining the accuracy by simply introducing a continuous-time anti-aliasing filter. Moreover, a transfer function form of the transmembrane current is presented that promote the use of sophisticated signal processing methods on these type of signals. Both continuous-time and discrete-time models are presented. We also address and analyze the implications of the finite length of the muscle fibers. Including this in the model is straightforward, owing to the convolutional form of the line source model, and is manifested by a simple transformation of the associated weighting function. AP modeling is discussed for the three different electrode models: the concentric needle electrode, the single fiber electrode, and the macro electrode. The presented model is suitable for modeling large motor units, where both accuracy and computational efficiency are important factors. To simplify the selection of the discretization interval, we derive what we call the cumulative cutoff frequency that provides an estimate of the required Nyquist frequency. PMID- 15543668 TI - Nitrite oxidation inhibition by hydroxylamine: experimental and model evaluation. AB - A proposed approach for biological nitrogen removal significantly reduces cost by reducing biomass production and carbon requirements via inhibition of nitrite oxidation (NO2- to NO3-). Batch experiments were conducted to examine the effect of hydroxylamine (HM) on nitrite oxidizers, ammonia oxidizers, and nitrite reducers. Hydroxylamine effect experiments were done at initial pH values of 7.4 8.4, nitrogen concentrations of 100 mg N/L, biomass concentrations of 100-400 mg VSS/L and HM dosages up to 43 mg/L. Nitrite oxidizer activity was completely inhibited by HM at dosages of 7.0 and 8.9 mg/L for pH values of 8.4 and 7.6, respectively. Relatively low HM concentrations (0.35-5.5 mg/L) can be used to completely inhibit nitrite oxidation, but do not significantly affect ammonia oxidizers and nitrite reducers. A model developed to describe the effect of pH on nitrite oxidation rate fits the data well (R2 = 0.89) with values for Vmax of 0.372 (mg N/mg VSS-hr), pH* of 7.72, and the inhibition constant Kh of 0.154. Incorporation of HM inhibition into the model provided a good fit to relative nitrite oxidation rate as a function of undissociated HM concentration (R2 = 0.80, Vmax = 0.028 mg N/mg VSS-hr, pH = 7.89, Kh = 0.302, a = 0.195, and Ki= 0.277 mg/L). PMID- 15543669 TI - AR: Sedated colonoscopy pt. killed while driving: judgment for G.I. Center & nurse reversed. PMID- 15543670 TI - Pharmacokinetics of recombinant human basic fibroblast growth factor in mice using a radioiodination method combined with SDS-PAGE and a sandwich enzyme linked immunosorbent assay. AB - Pharmacokinetics of recombinant human basic fibroblast growth factor (rhbFGF) in mice were studied by using a radioiodination method combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and a sandwich enzyme Linked immunosorbent assay (ELISA). RhbFGF concentration in serum were dertermined after iv administration of 2.5, 5 and 10 microg/kg of 125I-rhbFGF or rhbFGF. The AUCs were linearly correlated to doses. Concentrations in tissues, recoveries of 125I-rhbFGF in urine and feces were also analyzed following iv dose of 5 microg/kg. Pharmacokinetic profiles of radiolabelled and unlabelled rhbFGF were consistent in trend but exit certain differences. Mice cleared unlabeled rhbFGF markedly faster than their radiolabeled counterparts. PMID- 15543671 TI - Nurses' attitudes toward people with mental illnesses in a psychiatric hospital in Singapore. AB - A positive attitude toward people with mental illnesses is a necessary dimension of psychiatric nursing practice. Although studies have been conducted in Israel and Australia to examine the attitudes of nurses toward people with mental illnesses, no such study had been conducted in Singapore. This study explored the factors influencing nurses' attitudes toward and their involvement in caring for people with mental illnesses, and sought to establish the relevance and effects of mental health training on enhancing the nurses' positive attitudes. The study used a descriptive, self-administered questionnaire, which contained 24 statements scored on a Likert scale. Factors related to attitudes toward people with mental illnesses included age, professional qualification, years of psychiatric nursing experience, type of ward, and designation (i.e., position within the institution). The questionnaires were given to all of the local nurses working in a psychiatric hospital in Singapore, and 96% returned the questionnaires. Results showed that nurses with any of the following characteristics had a more positive attitude: age group of 31 to 50; professional qualification of advanced diploma in mental health nursing, nursing degree, or post-basic certificate; or more than 10 years of psychiatric nursing experience. In addition, nursing officers had a more positive attitude than staff nurses or assistant nurses, and nurses working in the short-stay wards had more positive attitudes than those working in the long-stay wards. PMID- 15543672 TI - Association between cerebrospinal fluid levels of asymmetric dimethyl-L-arginine, an endogenous inhibitor of endothelial nitric oxide synthase, and cerebral vasospasm in a primate model of subarachnoid hemorrhage. AB - OBJECT: Decreased availability of nitric oxide (NO) has been proposed to evoke delayed cerebral vasospasm after sub-arachnoid hemorrhage (SAH). Asymmetric dimethyl-L-arginine (ADMA) inhibits endothelial NO synthase (eNOS) and, therefore, may be responsible for decreased NO availability in cases of cerebral vasospasm. The goal of this study was to determine whether ADMA levels are associated with cerebral vasospasm in a primate model of SAH. METHODS: Twenty-two cynomolgus monkeys (six control animals and 16 with SAH) were used in this study. The levels of ADMA, L-arginine, L-citrulline, nitrites, and nitrates in cerebrospinal fluid (CSF) and serum were determined on Days 0, 7, 14, and 21 following onset of SAH. Cerebral arteriography was performed to assess the degree of vasospasm. Western blot analyses of the right and left middle cerebral arteries (MCAs) were performed to assess the expression of eNOS, type I protein arginine methyl transferase (PRMT1) and dimethylarginine dimethylaminohydrolase (DDAH2). Cerebrospinal fluid levels of ADMA remained unchanged in the control group (six animals) and in animals with SAH that did not have vasospasm (five animals; p = 0.17), but the levels increased in animals with vasospasm (11 animals) on Day 7 post-SAH (p < 0.01) and decreased on Days 14 through 21 (p < 0.05). Cerebrospinal fluid levels of ADMA correlated directly with the degree of vasospasm (correlation coefficient = 0.7, p = 0.0001; 95% confidence interval: 0.43-0.83). Levels of nitrite and nitrate as well as those of L-citrulline in CSF were decreased in animals with vasospasm. Furthermore, DDAH2 expression was attenuated in the right spastic MCA on Day 7 post-SAH, whereas eNOS and PRMT1 expression remained unchanged. CONCLUSIONS: Changes in the CSF levels of ADMA are associated with the development and resolution of vasospasm found on arteriograms after SAH. The results indicate that endogenous inhibition of eNOS by ADMA may be involved in the development of delayed cerebral vasospasm. Inhibition of ADMA production may provide a new therapeutic approach for cerebral vasospasm after SAH. PMID- 15543673 TI - Gender choice: is it playing God? PMID- 15543674 TI - Ethics & life's ending: an exchange. PMID- 15543675 TI - Human reproduction and rights of action and of recipience. PMID- 15543676 TI - Bioethics as a scientific enterprise. AB - The authors of this article attempt to look at the manner in which the field of bioethics has emerged to represent an entirely new science. The article does this by first looking at various developments in the field of medicine and science. Specifically, the authors point to the developments in pharmacology, biology, and in genetics, as the stimuli for the field of bioethics. Since these three fields of study directly deal with human beings and their respective environment, new developments in these respective areas of science have questioned what society has traditionally held concerning these two concepts. In an effort to help answer such questions, the field of bioethics has emerged. The authors argue that bioethics represents more than just a theoretical field. Instead, bioethics, as an entirely new science in itself, has the ability to provide answers to the question, and moral dilemmas that new developments in other sciences raise. The article attempts to show that science/medicine has direct links to proper therapy and procedures. The authors do this by making a direct connection between ethics and medicine, as Hippocrates understood. For Hippocrates, medicine and science should be distinguished from religion, but not from proper ethos. Therefore, the authors argue that the field of bioethics has gained its status as a scientific enterprise since it provides the ability for scientists and physicians to conduct their work in an ethos that aims to protect the human person. PMID- 15543677 TI - Human stem cell research. PMID- 15543678 TI - Is regulation of human cloning necessary? PMID- 15543679 TI - Exploiting abstract possibilities: a critique of the concept and practice of product patenting. AB - Developments in biotechnology and genomics have moved the issue of patenting scientific and technological inventions toward the center of interest. In particular, the patentability of genes of plants, animals, or humans and of genetically modified (parts of) living organisms has been discussed, and questioned, from various normative perspectives. This paper aims to contribute to this debate. For this purpose, it first explains a number of relevant aspects of the theory and practice of patenting. The focus is on a special and increasingly significant type of patents, namely product patents. The paper provides three general arguments against the concept and practice of product patenting. The first argument briefly considers the claim that patents are legitimate because they promote socially useful innovation. Against this claim, it is argued that product patents may hamper rather than promote such innovation. The second and main argument concludes that product patents are not adequately based on actual technological inventions, as they should be according to the usual criteria of patentability. The principal moral issue is that product patents tend to reward patentees for inventions they have not really made available. The final argument proposes a method for patenting the heat of the sun. Assuming that granting this patent will be generally considered absurd, the argument exposes a further, fundamental problem of the concept and practice of product patenting. PMID- 15543680 TI - The battle over stem cells. PMID- 15543681 TI - Opposing a proposed assisted-suicide law. PMID- 15543682 TI - Biotechnology and its public. AB - The author provides a perspective on three key areas of biotechnology: the public debate over hazards, the commercialization of the technology, and ethical issues that are arising in terms of application of genetic therapy to humans. He reviews the history of the recombinant DNA research debate beginning at Asilomar in 1975, when the molecular biologists called for a temporary moratorium on certain experiments, so the NIH might develop guidelines to govern this research. The author then characterizes the lobbying activities of the biologists against proposed regulation at the federal, state and local level. Despite the lobbying, he concludes that Congress did not enact legislation because of the exemplary behavior of the biologists, beginning with their meeting at Asilomar. The author next describes the impact commercialization of this technology will have on academic biologists who traditionally have not been involved in that process (as have chemists and physicists). He also comments on the ethics of biotechnology, concluding that the more powerful biology becomes, the more its uses and some control of those uses will be debated. PMID- 15543684 TI - Bile formation: do not ignore the role of plasma membrane-cytoskeleton linking proteins. PMID- 15543683 TI - A role for insect galectins in parasite survival. AB - Insect galectins are associated with embryonic development or immunity against pathogens. Here, we show that they can be exploited by parasites for survival in their insect hosts. PpGalec, a tandem repeat galectin expressed in the midgut of the sandfly Phlebotomus papatasi, is used by Leishmania major as a receptor for mediating specific binding to the insect midgut, an event crucial for parasite survival, and accounts for species-specific vector competence for the most widely distributed form of cutaneous leishmaniasis in the Old World. In addition, these studies demonstrate the feasibility of using midgut receptors for parasite ligands as target antigens for transmission-blocking vaccines. PMID- 15543685 TI - Life expectancy after traumatic brain injury. PMID- 15543687 TI - Integration of digitall-ceramic restorations. PMID- 15543686 TI - NMR assignment of the turtle prion protein fragment tPrP(121-225). PMID- 15543688 TI - Avoiding insufficient reduction in fixed prosthodontics. PMID- 15543689 TI - Cementation considerations for CAD/CAM all-ceramic restorations. PMID- 15543690 TI - Treatment selection for anterior endodontically involved teeth. AB - Innovations in material science and clinical techniques have expanded the number of treatment options available for nonvital anterior teeth. These options include the use of composite to fill the access opening with no additional treatment, crown placement, orthodontic extrusion, crown lengthening with or without orthodontic extrusion, dowel restorations with crown placement, and fixed bridge or implant therapy when extraction is necessary. Clinicians need to understand the benefits and limitations of each option in order to provide their patients with optimum function and aesthetics. Using case presentations, this article describes predictable approaches for the diagnosis, treatment planning, and restoration or replacement of endodontically treated teeth in the anterior region. PMID- 15543691 TI - The cutting edge: instrumentation and preparation concepts. PMID- 15543692 TI - The diagnostic tracing analysis--visualization by the numbers. AB - Aesthetic treatment planning for dentistry often involves nontraditional methods of extracting information from clinical data. Being able to visualize the final treatment solution accurately in the initial diagnostic stages of treatment planning benefits both the clinician and patient. For the dentist it helps define the final treatment plan, allows for identification of unique nuances of the particular case, and contributes to the ease of fabrication of three dimensional treatment aids. It is also important for the dentist to verify that the patient's expectations of treatment are consistent with clinical reality. This article will describe the diagnostic tracing analysis, a tool that can help the visualization process. PMID- 15543693 TI - Placement of an implant-stabilized complete denture: a case report. PMID- 15543694 TI - Use of a zirconia framework and pressed veneering ceramics to replicate natural interproximal contours. PMID- 15543695 TI - New trends in polishing direct resin composites. PMID- 15543696 TI - Rotated palatal flaps: A functional and aesthetic solution in endentulous sites. AB - Resorption of alveolar process generally occurs following tooth extraction, and this process may cause a variety of jaw deformities. Bone deformities in the aesthetic zone impair the appearance, quality, and prognosis of the definitive restoration. The rotated palatal flap is a surgical modality that can be used to reduce or prevent alveolar resorption and assist in the repairing of exisiting defects. This presentation demonstrates the application of the rotated palatal flap in a variety of clinical cases as a means of addressing such resorption. PMID- 15543697 TI - HIPAA training requirements. AB - The addition of security standard training requirements increases the complexity of HIPAA implementation and ongoing program management. In comparison to the expectations of the Privacy Rule, security elements are more numerous and complicated--they also represent new territory for the radiology manager, who may be encountering a new language and unfamiliar technological terminology. The deadline for compliance with the Security Standard is April 21, 2005. It is, therefore, important to review training expectations as part of overall HIPAA compliance preparations. PMID- 15543698 TI - The demands of the workplace. PMID- 15543699 TI - Paperless medical records: measuring success. AB - North Shore Magnetic Imaging Center (NSMIC) underwent a major transformation of the patient process through an 18-month "Reinvention Project." The project began in October 2002, with an assessment of systems in place. A complete review of each stage of the patient process--scheduling, registration, insurance verification, screening, scanning, transcription, and billing--resulted in the discovery that the paperwork for a single patient examination could go though as many as 20 sets of hands. The project was supported by the formation of an internal team comprised of staff members from all departments (support, patient accounts, and technical), radiologists, and members of the center's senior management team. The team had 2 goals: increase the level of patient care, and create a paperless environment. External teams were formed to address specific areas targeted to support the process. The transformation for all involved- patients, staff, radiologists, and referring physicians--has proved to be very positive. The work, however, was not finished. Upon the project's completion, NSMIC recognized the importance of identifying successes and areas for improvement. These included ongoing reviews of the project's impact on all stakeholders and looking for new technologies and programs to enhance the new systems in place. There are plans for a project "sequel." Strategies are being developed for "Reinvention, Phase II." Elements of these strategies include enhancement of the scheduling programs to create more checks and balances for the staff and investigating an online scheduling option for the center's referring physicians. The purchase and implementation of a voice recognition system--tabled during Phase I--is scheduled for the first quarter of 2005. PMID- 15543700 TI - PACS strategy for imaging centers. AB - Picture archiving and communications systems (PACS) have been available in imaging centers for many years, but they often were less functional, were not well integrated into patient information systems, and lacked the network backbone to implement a system. As modalities are replaced and technology improves, the ability and time for an imaging center to acquire, integrate, and utilize PACS has arrived. However, each imaging center must determine why it should invest in PACS. A business plan is the fundamental need. Each imaging center must understand its target market, growth rate, and staffing plans. Additional considerations lie in current and future modality availability, the need for offsite delivery of images and reports, and the potential need for remote transmission of images. These issues must be identified and prioritized. A multidisciplinary team is essential. The most successful PACS implementation begins with complete involvement from all levels. The team should be comprised of people with complementary skills who are committed to a common purpose, set of performance goals, and approach for which they hold themselves mutually accountable. The team must jointly decide on the project's objectives. These objectives fall under 4 categories: clinical, service, financial, and performance. PACS must be considered a tool to help accomplish each objective. The imaging center must determine its top priorities, then translate them into a technology "wish list." The center can then list those pieces of technology that are most important and prioritize them. There are even more considerations for connecting multiple imaging centers. The team must create a comprehensive request for proposal (RFP) and determine the vendors that will receive the document. Once the RFP responses have been received and the vendor has been selected, an effective training plan must be executed. Training plans should be competency based, ensuring comfort and competency among all staff. Upon the project's completion, it is essential for the imaging center to evaluate PACS' effect upon its services and relationships with patients, staff, and referring physicians. Marketing and demonstrating the latest technology can positively impact all these areas. PMID- 15543701 TI - New healthcare trends that will impact radiology. PMID- 15543702 TI - Providing support to family members. AB - Providing recognition and simple information can be powerful and reassure a family member who is anxious and worried about an ill relative. While "answers" concerning the procedure are often not available, providing information that is available--the locations of the rest rooms and coffee shop; approximately how long the procedure will take; where to wait; and the process involved in the radiological procedure, reading and getting the results--can give some feeling of control, and thus reduce feelings of powerlessness. Most importantly, family members who are recognized and included with the patient in the treatment process will be reassured of the competency of the staff and gain hopefulness about the outcome of the diagnostic and treatment experience. And providing hope in the healthcare setting is a radiology professional's most important job. PMID- 15543703 TI - Management and art. PMID- 15543704 TI - Women's health issues require unique approach. PMID- 15543705 TI - Project primer. PMID- 15543706 TI - Managment imaging operations: tracking paid hours/volume/budget performance. PMID- 15543707 TI - Current perspectives on CT technology. AB - In 2004, the worldwide sales for computed tomography (CT) scanners will be approximately dollars 2.95 billion. Multislice scanning has had a profound effect on radiology and, subsequently, on diagnostic imaging. When planning a purchase for conventional CT imaging, high quality is always a prime consideration. Additionally, it appears to be almost mandatory to purchase a multislice system just to have the security of remaining technologically current, and capable, for the next 5 to 7 years. When buying a multislice system, it is essential to have the appropriate applications and technology--including a picture archiving and communication system (PACS) and workstations--that can handle the proliferation of data associated with new CT systems. Coverage is emerging as the most significant factor (over number of slices and speed) when considering CT technology. Coverage becomes more important when considering studies of organ function over time--such as cardiac function or perfusion of blood in a stroke imaging, etc.--rather than just morphology. A major challenge is the ability of being able to store all the data that result from a scanning procedure. In some cases,this could mean storing literally thousands of images per exam. In that regard, the information technology (IT) environment in the average hospital is not always equipped to handle the intensity of data from a high-end scanner. Cardiac imaging probably represents the greatest impact of CT on diagnostic medicine, while molecular imaging shows the greatest potential for diagnostic imaging, including CT. PMID- 15543708 TI - [Resource support as the basis of projects for the prevention of violence and addiction]. AB - Lastingly effective projects for the prevention of violence and addiction must ask which resources adolescents require for coping with their lives. A central criteria for a successful life is the creation of coherence in one's inner life and thus the basis for becoming capable of functioning in society. In earlier social eras, the readiness to take on ready-made identity packages was the central criteria for coping with life. Today, this depends on individual adaptation and identity work, and thus the capability for self-organisation, for "intentionality" or "embedding". Children and young people need "free spaces" in their worlds, in order to be able to outline themselves and to act formatively upon their everyday environment. The future prospects of adolescents depend upon their options for learning the "craft of freedom". Sustainable key qualifications for coping with life in globalised, digital capitalism must grasp education as a hard-headed process in which the subject's capacity for self-organisation is to be given optimal encouragement, so that the patchwork of their own identity can succeed as a self-determined creative project. PMID- 15543709 TI - [Change to the power of three--counselling for adolescents in a society which causes uncertainty]. AB - The conditions for becoming adult have changed perceptibly for young people in the course of social modernization processes. Adolescents find themselves in increasingly complex and contradictory areas of life which offer them more choices of action, but also require of them more self-organisation and more decision-making processes in uncertain conditions. The adolescents primarily at risk of failing due to the changes in learning- and development-related tasks are those denied access to necessary coping resources. Owing to the profound effects of social change (for example, changed forms of family life, increased educational pressure) it can generally be assumed that young people have an intensified counselling need. Against this background, the present paper examines which new requirements for action arise in the institutionalized counselling of adolescents. In regard to this, hints are provided in the results of an evaluation study by the present author, within the scope of which adolescents were interviewed about their counselling experiences. Finally, counselling approaches are described which can contribute to facilitating access for young clients, and to structuring counselling in a more suitable way for adolescents (peer counselling). PMID- 15543710 TI - [Counselling with modern expressions of mystic pictures]. AB - The picture-worlds of children in the 21th century are not only evoked anymore by fairytales of the Brothers Grimm, but also by modern audio-visual stories (in books, comics, mangas, swap cards, films, videos and computer games). In psychological counselling and psychotherapy of children and adolescents we like to use such picture-worlds, modern myths and metaphors, to tie up with the experiences of the children and to kick start corresponding processes of maturation. Consultants can not only use their own treasures of myths remembering their own childhood (reading), but should also be familiar with the modern picture-worlds common to children in order to develop images which are shared by both counsellor and child, and should be capable of understanding the meaning of the characters and their actions. Some narratives and forms will be presented here in relation to developmental psychology. PMID- 15543711 TI - [Virtual counselling: online help for adolescents]. AB - The internet provides a new platform for communication especially used by teenagers and young adults. Professional counselling in this medium offers the chance to reach especially those individuals perhaps not reachable through existing counselling services. With this in mind, the internet portal bke (youth counselling of the Federal conference for Educational Counselling) was inaugurated in October 2000. It offers the following services: Group Chat, Discussion Forum and E-Mail Counselling. The experience made in this project phase indicates that professional counselling in this medium can reach individuals directly, who would otherwise not or only with difficulty be reached by the ordinary counselling services on offer. This is due to the fact that the internet is easily accessible, offers anonymity and therefore presents no noteworthy access barrier to overcome. This seems to apply especially to traumatised teenagers and young adults. Online counselling offers great opportunities exactly in these cases. The special task of a counsellor is to grasp this opportunity whilst at the same time having an awareness of the mediums limitations. PMID- 15543712 TI - Topical retinoids in the management of acne: the best path to clear results. PMID- 15543713 TI - Topical tazarotene: The BEST (balancing efficacy, speed, and tolerability) in acne trial. AB - Results of clinical trials have shown that tazarotene cream used as monotherapy effectively reduces the number of both inflammatory and noninflammatory lesions in patients with acne vulgaris. The efficacy of tazarotene cream in clinical practice, however, has not been reported. To evaluate the benefits of tazarotene cream in everyday clinical practice, an uncontrolled observational study was conducted in a large population of patients with facial acne vulgaris; 243 investigators enrolled 1118 patients for treatment with once-daily tazarotene 0. 1% cream as monotherapy, or in combination with other agents, for up to 12 weeks. Tazarotene efficacy was well rated by both investigators and patients and had a rapid onset of action. Reduction in comedone counts was greatest when tazarotene 0.1% was used as first-line therapy. The incidence of adverse events (almost exclusively peeling, erythema, dryness, or burning) was low and typically declined with continued use of the medication. The combination of tazarotene 0.1% cream and clindamycin-benzoyl peroxide was the best tolerated of all the treatment regimens. Patients and investigators reported high levels of treatment satisfaction with topical tazarotene 0.1% cream. PMID- 15543714 TI - Meta-analysis of topical tazarotene in the treatment of mild to moderate acne. AB - Topical tazarotene has shown superior efficacy over other topical retinoids, including adapalene and tretinoin, in the treatment of mild to moderate acne. A meta-analysis of data from 6 multicenter, double-blind, randomized comparative trials was performed to determine how patient characteristics influence the efficacy and tolerability of topical tazarotene. Data on 468 patients who used tazarotene 0.1% gel or cream once daily for 12 weeks were collected. Topical tazarotene was effective and well tolerated, regardless of patients' acne severity, skin type, sex, or ethnicity. In general, 'using the cream formulation rather than the gel formulation optimized tolerability. These results indicate that topical tazarotene 0.1% gel and cream are efficacious and well-tolerated treatment options for clearing acne vulgaris across a broad range of patients. PMID- 15543715 TI - Topical tazarotene in acne vulgaris: treatment approaches. AB - Topical tazarotene is effective and well tolerated in the treatment of acne vulgaris and has been used successfully in diverse patient populations. However, because factors such as patient skin type and climate may influence its optimal use and frequency of application, treatment approaches often differ according to the populations in which tazarotene is used. The authors review their prescribing practices and special considerations for women, African Americans, patients living in dry climates, adolescents, and Asian Americans. PMID- 15543716 TI - The flu shot fiasco. PMID- 15543717 TI - Scary strains. PMID- 15543718 TI - Arsenic and lead leach out of popular fertilizer. PMID- 15543719 TI - Neglected forms of phosphorus play important role. PMID- 15543720 TI - Eutrophication implicated in deformed frogs. PMID- 15543722 TI - Are environmental exemptions for the U.S. military justified? PMID- 15543721 TI - PBDEs and the environmental intervention time lag. PMID- 15543723 TI - Targeting methane emissions. PMID- 15543724 TI - Scrutinizing pharmaceuticals and personal care products in wastewater treatment. PMID- 15543725 TI - Clean Water Act assessment processes in relation to changing U.S. Environmental Protection Agency management strategies. AB - During the 1970s the U.S. Environmental Protection Agency (EPA) devised a multiscale system of basin planning and regional implementation that encouraged a balanced mixture of monitoring and modeling-based assessments. By the 1980s, this goal had not been achieved. Modeling and monitoring assessment approaches became largely decoupled. To a significant degree, modeling was viewed as too inaccurate to handle issues such as setting permit limits involving toxics. During the 1980s, EPA also encouraged the idea that monitoring approaches were adequate to document water quality problems, guide the development of management plans, and demonstrate the achievement of management goals. By the late 1990s, large numbers of waters listed under the Clean Water Act's Total Maximum Daily Load (TMDL) provisions showed the widespread nature of pollutant concerns, but the uneven nature of the listing information also revealed fundamental problems in the ability of state monitoring programs to achieve credible and comprehensive assessments. Statistics are presented from the 1998 and the most current publicly available 2000 baseline periods showing the limitations in the scope of state assessments. There are significant opportunities for the increased use of relatively simple modeling systems that can be flexibly implemented over a variety of spatial scales. In addition to conventional modeling frameworks, the value of bioassessment monitoring techniques is stressed. Bioassessment indicators can often be combined with landscape modeling methods, as well as analyses from conventional modeling outputs, to help target small area monitoring by use of tiered approaches. These findings underscore the value of integrated monitoring and modeling approaches to build properly balanced assessment systems. PMID- 15543726 TI - Emissions of polycyclic aromatic hydrocarbons from batch hot mix asphalt plants. AB - This study was set out to assess the characteristics of polycyclic aromatic hydrocarbon (PAH) emissions from batch hot mix asphalt (HMA) plants and PAH removal efficiencies associated with their installed air pollution control devices. Field samplings were conducted on six randomly selected batch HMA plants. For each selected plant, stack flue gas samples were collected from both stacks of the batch mixer (n = 5) and the preheating boiler (n = 5), respectively. PAH samples were also collected from the field to assess PAHs that were directly emitted from the discharging chute (n = 3). To assess PAH removal efficiencies of the installed air pollution control devices, PAH contents in both cyclone fly ash (n=3) and bag filter fly ash (n = 3) were analyzed. Results show that the total PAH concentration (mean; RSD) in the stack flue gas of the batch mixer (354 microg/Nm3; 78.5%) was higher than that emitted from the discharging chute (107 microg/Nm3; 70.1%) and that in the stack flue gas of the preheating boiler (83.7 microg/Nm3; 77.6%). But the total BaPeq concentration of that emitted from the discharging chute (0.950 microg/Nm3; 84.4%) was higher than contained in the stack flue gas of the batch mixer (0.629 microg/Nm3; 86.8%) and the stack flue gas of the preheating boiler (= 0.112 microg/Nm3; 80.3%). The mean total PAH emission factor for all selected batch mix plants (= 139 mg/ton x product) was much higher than that reported by U.S. EPA for the drum mix asphalt plant (range = 11.8-79.0 mg/ton x product). We found the overall removal efficiency of the installed air pollution control devices (i.e., cyclone + bag filter) on total PAHs and total BaPeq were 22.1% and 93.7%, respectively. This implies that the installed air pollution control devices, although they have a very limited effect on the removal of total PAHs, do significantly reduce the carcinogenic potencies associated with PAH emissions from batch HMA plants. PMID- 15543727 TI - Speciation of PM10 sources of airborne nonferrous metals within the 3-km zone of lead/zinc smelters. AB - The purpose of this study was to estimate the speciation of PM10 sources of airborne Pb, Zn, and Cd metals (PM10 is an aerosol standard of aerodynamic diameter less than 10 microm.) in the atmosphere of a 3 km zone surrounding lead/zinc facilities in operation for a century. Many powdered samples were collected in stacks of working units (grilling, furnace, and refinery), outdoor storages (ores, recycled materials), surrounding waste slag (4 Mt), and polluted topsoils (3 km). PM10 samples were generated from the raw powders by using artificial resuspension and collection devices. The bulk PM10 multielemental analyses were determined by inductively coupled plasma-atomic emission spectrometry (ICP-AES). The proportions in mass of Pb (50%), Zn (40%), and Cd (1%) contents and associated metals (traces) reach the proportions of corresponding raw powdered samples of ores, recycled materials, and fumesize emissions of plants without specific enrichment. In contrast, Pb (8%) and Zn (15%) contents of PM10 of slag deposit were found to be markedly higher than those of raw dust, Pb (4%), and Zn (9%), respectively. In the same way, Pb (0.18%), Zn (0.20%), and Cd (0.004%) were enriched by 1.7, 2.1, and 2.3 times, respectively, in PM10 as compared with raw top-soil corresponding values. X-ray wavelength dispersive electron-microprobe (EM-WDS) microanalysis did not indicate well-defined phases or simple stoichiometries of all the PM10 samples atthe level of the spatial resolution (1 microm3). X-ray photoelectron spectroscopy (XPS) indicated that minor elements such as Cd, Hg, and C are more concentrated on the particle surface than in the bulk of PM10 generated by the smelting processes. (XPS) provided also the average speciation of the surface of PM10; Pb is mainly represented as PbSO4, Zn as ZnS, and Cd as CdS or CdSO4, and small amounts of coke were also detected. The speciation of bulk PM10 crystallized compounds was deduced from XRD diffractograms with a raw estimation of the relative quantities. PbS and ZnS were found to be the major phases in PM10 generated by the smelting facilities with PbSO4, PbSO4 x PbO, PbSO4 x 4PbO, Pb metal, and ZnO as minor phases. The slag waste PM10 was found to contain some amounts of PbCO3, PbSO4 x PbO, and ZnFe2O4 phases. The large heterogeneity at the level of the individual particle generates severe overlap of chemical information even at the microm scale using electron microprobe (WDS) and Raman microprobe techniques. Fortunately, scanning Raman microspectrometry combined with SIMPle-to-use Interactive Self-modeling Mixture Analysis (SIMPLISMA) performed the PM10 speciation at the level of individual particles. The speciation of major Pb, Zn, and Cd compounds of PM10 stack emissions and wind blown dust of ores and recycled materials were found to be PbSO4, PbSO4 x PbO, PbSO4 x 4PbO, PbO, metallic Pb, ZnS, ZnO, and CdS. The PM10 dust of slag waste was found to contain PbCO3, Pb(OH)2 x 2PbCO3, PbSO4 x PbO, and ZnS, while PM10-bound Pb, Zn of the top-soils contain Pb5(PO4)3Cl, ZnFe2O4 as well as Pb(II) and Zn(II) compounds adsorbed on Fe(III) oxides and in association with clays. PMID- 15543728 TI - Annual variations of pesticide concentrations in Great Lakes precipitation. AB - Twenty pesticides and related analytes were measured in 28-day integrated precipitation samples from five U.S. sites in the Integrated Atmospheric Deposition Network (IADN) between 1997 and 2002. Consistent, significant decreases in concentration as a function of time were observed only for p,p'-DDE and p,p'-DDD, while increases in beta-HCH were observed at all sites. Significant annual variations were observed for most analytes at each site with higher concentrations in the summer for current-use pesticides (endosulfan and gamma HCH) and peaks in the winter for most others. The increased concentrations in the winter are likely the result of the increased scavenging efficiency of snow compared to rain and, for some analytes, higher concentrations in the particulate phase during winter. These seasonal differences appear to account for a large portion of the observed variability in pesticide concentrations in precipitation samples. PMID- 15543729 TI - Biota--sediment accumulation factors for polychlorinated biphenyls, dibenzo-p dioxins, and dibenzofurans in southern Lake Michigan lake trout (Salvelinus namaycush). AB - A set of high-quality, age-specific biota-sediment accumulation factors (BSAFs) for polychlorinated biphenyls (PCBs), dibenzo-p-dioxins (PCDDs), and dibenzofurans (PCDFs) have been determined from concentrations measured with high resolution gas chromatography/high-resolution mass spectrometry, by use of the 13C isotopic dilution technique, in lake trout and surficial (0-2 cm) sediment samples from southern Lake Michigan. BSAFs ranged from <0.1 to 18 for PCBs and from <0.001 to 0.32 for PCDDs and PCDFs detected in the fish. PCBs with zero or one chlorine in an ortho position had smaller BSAFs than other PCBs. PCDDs and PCDFs with chlorines at the 2,3,7,8-positions had larger BSAFs than most other PCDDs and PCDFs. The fidelity of the relative bioaccumulation potential data between independent lake trout samples, within and among age classes, suggests that differences in slight rates of net metabolism in the food chain are important and contribute to the apparent differences in BSAFs, not only for PCDDs and PCDFs but also possibly for some PCBs. A complicating factor for non-ortho- and mono-ortho-PCBs is the uncertain contribution of enhanced affinity for black carbon (and possibly volatility) acting in concert with metabolism to reduce measured BSAFs for lake trout. On the basis of the association between chemicals with apparent slight rates of metabolism and measured dioxin-like toxicity, several PCDFs with similar measured BSAFs but unknown toxicity may be candidates for toxicity testing. PMID- 15543730 TI - Polybrominated diphenyl ethers contamination of United States food. AB - Elevated levels of polybrominated diphenyl ethers (PBDEs), a type of brominated flame retardant, were recently detected in U.S. nursing mothers' milk. These halogenated compounds chemically and toxicologically resemble others such as polychlorinated biphenyls (PCBs), whose route of intake is almost exclusively through food of animal origin. This study is the first to report the levels of PBDEs in U.S. foods in a market basket survey of 30 food types (total of 32 food samples) from three major supermarket chains in Dallas, TX. Food samples were almost exclusively foods of animal origin: meat, fish, and dairy products. Thirteen PBDE congeners were measured for each sample. Levels were then compared to existing PBDE food studies from other countries where available. In this study, levels of PBDEs are highest in fish, then meat, and lowest in dairy products; median levels were 1725 (range 8.5-3078), 283 (range 0.9-679), and 31.5 (0.2-1373), parts per trillion (ppt), or pg/g, wet weight, respectively. Nonfat milk did not have any detectable PBDE levels. In fish, PBDE congener 47 (2,2',4,4'-tetraBDE) contributes up to 70% of the total PBDEs, followed by congeners 100 (2,2',4,4',6) and 99 (2,2',4,4',5). In meat congener 99 predominates, followed by 47. In dairy, BDE 47 predominates followed by 99. U.S. food PBDE levels measured in this study are higher than reported in two other published market based studies from Spain and Japan. Although these findings are preliminary and will be updated with analyses of new samples, they suggest that food is a major route of intake for PBDEs. PMID- 15543734 TI - Chloroacetanilide herbicide metabolites in Wisconsin groundwater: 2001 survey results. AB - A survey of agricultural chemicals in Wisconsin groundwater was conducted between October 2000 and April 2001 to obtain a current picture of agricultural chemicals in groundwater used for private drinking water. A stratified, random sampling procedure was used to select 336 sampling locations. Water from private drinking water wells randomly selected from within the 336 sampling locations was analyzed for 18 compounds including herbicides, herbicide metabolites, and nitrate. This report focuses on the frequency and concentration of chloroacetanilide herbicides and their metabolites. Analysis of data resulted in an estimated proportion of 38+/-5.0% of wells that contained detectable levels of a herbicide or herbicide metabolite. The most commonly detected compound was alachlor ESA with a proportion estimate of 28+/-4.6%. Other detected compounds in order of prevalence were metolachlor ESA, metolachlor OA, alachlor OA, acetochlor ESA, and parent alachlor. Estimates of the mean concentration for the detects ranged from 0.15+/ 0.082 microg/L for acetochlor ESA to 1.8+/-0.60 microg/L for alachlor OA. Water quality standards have not been developed for these chloroacetanilide herbicide metabolites. The results of this survey emphasize the need for toxicological assessments of herbicide metabolite compounds and establishment of water quality standards at the state and federal levels. PMID- 15543738 TI - Accumulation of arsenic in drinking water distribution systems. AB - The tendency for iron solid surfaces to adsorb arsenic is well-known and has become the basis for several drinking water treatment approaches that remove arsenic. It is reasonable to assume that iron-based solids, such as corrosion deposits present in drinking water distribution systems, have similar adsorptive properties and could therefore concentrate arsenic and potentially re-release it into the distribution system. The arsenic composition of solids collected from drinking water distribution systems (pipe sections and hydrant flush solids), where the waters had measurable amounts of arsenic in their treated water, were determined. The elemental composition and mineralogy of 67 solid samples collected from 15 drinking water utilities located in Ohio (7), Michigan (7), and Indiana (1) were also determined. The arsenic content of these solids ranged from 10 to 13 650 microg of As/g of solid (as high as 1.37 wt %), and the major element of most solids was iron. Significant amounts of arsenic were even found in solids from systems that were exposed to relatively low concentrations of arsenic (<10 microg/L) in the water. PMID- 15543735 TI - Radiocarbon apportionment of fossil versus biofuel combustion sources of polycyclic aromatic hydrocarbons in the Stockholm metropolitan area. AB - Source-diagnostic markers and the isotopic composition of polycyclic aromatic hydrocarbons (PAHs) were examined in surface sediments from the greater Stockholm waterways to deduce the contribution from biomass sources to the environmental PAH load. The summed concentration of 20 PAHs ranged from 0.8 to 45.1 microg/g (dry weight) and exhibited a steep decline with increasing distance from the city center evidencing that sources within the metropolitan area of Stockholm dominate its PAH burden. Several diagnostic PAH ratios indicated an overwhelming predominance of pyrogenic sources over the petrogenic ones, while retene and 1,7 dimethylphenanthrene were unable to correctly evaluate the contribution from biomass combustion. The stable carbon isotope composition (delta13C) of individual PAHs ranged from -24.8 to -27.0% but also was proved inefficient to discriminate between different types of fuels due to the overlapping signals in various sources. The delta14C values of PAHs ranged between -550.4 and -934.1%, indicating a clear predominance of fossil fuel sources. By using an isotopic mass balance approach, we estimated that on average 17+/-9% of PAHs derived from biomass combustion. This radiocarbon apportionment, in conjunction with detailed energy statistics for the Stockholm region, revealed that the ambient PAH burden is roughly similar, per unit energy produced, from fossil fuels and biofuels. Societies' shifting energy policies toward a larger reliance on biofuels may thus not lead to further deterioration of air quality and respiratory ailments for the urban population. PMID- 15543732 TI - Trace metal and major ion inputs into the Olentangy River from an urban storm sewer. AB - Trace metal clean techniques were used to sample and analyze the input of dissolved trace metals, major ions, and dissolved organic carbon (DOC) from a storm sewer along an urban highway in Columbus, OH. The outfall, draining a 3.6 ha sewershed with 100% impermeable surface area, discharges into the Olentangy River. Dissolved Pb (average concentration of 3 nM) and dissolved Zn (average concentration of 127 nM) were found to be much lower in concentration than reported in previous investigations of dissolved metals in urban stormwater runoff. Average concentrations of dissolved Cr (1 microM), Ni (0.087 microM), and Cu (0.33 microM) were similar to those reported in previous studies. The storm sewer is shown to be a significant source of V, Ni, and Zn to the river. The outfall is also a significant source of Na, NH4, Cl, and DOC. The storm sewer input is depleted in NO2 and NO3 as compared to the river, reflecting the highly agricultural land use of the watershed upstream of the sewershed. Input from the storm sewer is also depleted, as compared to the river, with respect to dissolved Mg, Sr, and U with probable sources in the limestone/shale bedrock and glacial till-derived soils in the watershed. PMID- 15543739 TI - Structure elucidation and characterization of polychlorinated biphenyl carboxylic acids as major constituents of chromophoric dissolved organic matter in seawater. AB - Chromophoric or colored dissolved organic matter (CDOM) is one of the principal light adsorbing components of seawater, particularly in the ultraviolet, where it attenuates over 90% of downwelling ultraviolet radiation. In highly productive coastal regions and throughout most of the global ocean, in situ biological production is the major source of CDOM. However, little is known about CDOM composition on the molecular level, and there are only a few reports that link CDOM composition to autochthonous biological sources. Here we report the isolation and characterization of CDOM components from one coastal and two open ocean sites. Each sample contains a complex mixture of light absorbing (300-400 nm) components, including 2,4-dichlorobenzoic acid and a suite of novel, polychlorinated biphenyl carboxylic acids that closely resemble polychlorinated biphenyls (PCBs) of anthropogenic origin. However, the global inventory and isomer distribution of dissolved chlorinated aromatic acids suggest they are derived from in situ biological production rather than anthropogenic contaminants. These novel chlorinated aromatic acids account for a significant amount of CDOM adsorption in the ultraviolet. PMID- 15543733 TI - VOCs in shallow groundwater in new residential/commercial areas of the United States. AB - The quality of shallow groundwater in urban areas was investigated by sampling 518 monitoring wells between 1996 and 2002 as part of the National Water-Quality Assessment Program of the U.S. Geological Survey. Well networks were installed primarily in new residential/commercial areas less than about 30 years old (17 studies) and in small towns (2 studies) by randomly locating as many as 30 monitoring wells in each study area. The median well depth was 10 m. Based on samples with age-date information, almost all groundwater was recharged after 1950. Samples were analyzed for 53 volatile organic compounds (VOCs). Concentrations ranged from about 0.001 to 1000 microg/L (median 0.04), with less than 1% of the samples exceeding a Maximum Contamination Level or Drinking Water Advisory established by the U.S. Environmental Protection Agency. Using uncensored concentration data, at least one VOC was detected in 88% of the samples, and at least two VOCs were detected in 69% of the samples. Chloroform, toluene, and perchloroethene were the three most frequently detected VOCs. Dissolved oxygen concentration, estimated recharge index, and land-use were significant variables in logistic regression models that explained the presence of the commonly detected VOCs. Dissolved oxygen concentration was the most important explanatory variable in logistic regression models for 6 of the 14 most frequently detected VOCs. Bromodichloromethane, chloroform, and 1,1,1 trichloroethane had a positive correlation with dissolved oxygen; in contrast, dichloroethane, benzene, and toluene had a negative correlation with dissolved oxygen. PMID- 15543736 TI - Indoor and outdoor polycyclic aromatic hydrocarbons in residences surrounding a Soderberg aluminum smelter in Canada. AB - Ambient air in 18 residences surrounding an aluminum smelter were sampled to study the relationship between indoor and outdoor polycyclic aromatic hydrocarbons (PAHs). Objectives of the study were to quantify the indoor distribution of PAHs, indoor/outdoor (I/O) concentration ratios, and the relationship among PAH compounds. Correlation coefficients inside residences suggested an indoor source of 2-3 ring PAHs and an external source of 4-6 ring PAHs. The I/O ratios of 4-6 ring PAHs for homes without any substantial indoor sources were below unity, indicating that the presence of these PAHs was attributable to the aluminum smelter. Least squares linear regression of the coupled measurements without indoor sources of 5-6 ring PAHs resulted in average infiltration efficiencies (P(PAH)) of 0.49, 0.20, and 0.47 for benzo[a]pyrene, benzo[k]fluoranthene, and benzo[g,h,i]perylene, respectively. These P(PAH) values suggest that simultaneous measurements of indoor and outdoor concentrations of PAHs > 4 rings predominantly associated with the fine fraction of particulate matter could provide useful estimates of particle infiltration efficiency. Overall, study results indicate that when an industrial facility is the main source of outdoor 4-6 ring PAHs, the contribution of facility emissions may greatly exceed indoor sources in nonsmoking residences. PMID- 15543731 TI - Passive sampling survey of polybrominated diphenyl ether flame retardants in indoor and outdoor air in Ottawa, Canada: implications for sources and exposure. AB - The polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants in plastics of soft furnishings, TV sets and computers, and insulation in the indoor environment. The penta-BDEs--now banned in most parts of Europe but still used in North America--are additive flame retardants that may be released to the indoor environment via volatilization or as dusts. In this study, to investigate general population PBDE exposure, air was sampled in 74 randomly selected homes in Ottawa, Canada and at seven outdoor sites during the winter of 2002--3, using polyurethane foam (PUF) disk passive air samplers. The passive sampling rate (2.5 m3 day(-1)) was determined through a pilot study employing active and passive samplers side-by-side at selected indoor locations. Indoor air concentrations of PBDEs were log-normally distributed with a geometric mean of 120 pg m(-3) and a median of 100 pg m(-3), approximately 50 times higher than the range of outdoor air concentrations (<0.1-4.4 pg m(-3)). The maximum daily human exposure via the inhalation pathway based on median PBDE levels found in this survey was estimated to be 1.9 ng day(-1) (female) and 2.0 ng day(-1) (male), representing 4.1% (f) and 4.4% (m) of overall daily intake. PMID- 15543743 TI - Arsenic and lead leaching from the waste derived fertilizer ironite. AB - The toxicity characteristic leaching procedure (TCLP) and the synthetic precipitation leaching procedure (SPLP) were performed on commercially purchased samples of the waste-derived soil amendment marketed as Ironite. Ten samples of the 1-0-0 grade (the most widely available in Florida) were tested. Two samples of the 12-10-10 grade and three samples of the 6-2-1 grade (a liquid version) were tested as well. TCLP leachate concentrations from the 1-0-0 grade samples ranged from 5.0 to 8.0 mg L(-1) for lead and 2.2 to 4.8 mg L(-1) for arsenic. SPLP concentrations from the 1-0-0 samples ranged from 0.62 to 3.1 mg L(-1) for lead and 1.9 to 8.2 mg L(-1) for arsenic. All of the 1-0-0 grade samples exceeded the U.S. hazardous waste toxicity characteristic (TC) limit for lead (5 mg L( 1)), while five of the 10 SPLP samples exceeded the TC limit for arsenic (5 mg L( 1)). The greater arsenic leachability in the SPLP relative to the TCLP was determined to be a result of lower pH conditions in the SPLP. A composite sample of the 1-0-0 grade was found to leach much greater concentrations of both arsenic and lead using California's waste extraction test (WET). Lead leachate concentrations were much lower in the two 12-10-10 samples (0.03 mg L(-1) or less); arsenic concentrations in these leachates (both TCLP and SPLP) exceeded 5 mg L(-1). None of the 6-2-1 samples contained lead or arsenic above TC limits. An experiment performed on the 1-0-0 grade which examined leachability as a function of pH found that at pH values in the range of what is encountered in the human digestive system (pH 4.0 to 1.5) lead leached 2-36% of its initial content, and arsenic leached 1-6% of its initial content. A simple gastric acid leaching experiment found 83 and 37% of the lead and arsenic present to leach, respectively. PMID- 15543737 TI - Transport of gas-phase polycyclic aromatic hydrocarbons to the Venice lagoon. AB - Concentrations of gas-phase polycyclic aromatic hydrocarbons (PAHs) were studied over one year at two sites of the Venice lagoon (designated Marine and Industrial) and at a mainland station (designated Rural) in Italy. Average sigmaPAH concentrations, calculated as sum of 16 PAHs, at Marine are about three and five times lower than those at Industrial and Rural, respectively. The seasonal trends, the temperature-PAH relationship, and principal component analysis indicate that at Industrial and Marine sites several local sources (vehicle and industrial emissions, etc.) could be the PAH sources in the warmer months, whereas in the colder months the main PAH sources could alternate between vehicle emissions and residential heating. At Rural the main PAH sources are: vehicle emissions in the spring and autumn; vehicle emissions, field burning, and wood combustion in the summer; and vehicle emissions and fuel consumption for residential heating in the winter. To evaluate the contribution from different sources to the Venice Lagoon air, horizontal fluxes of PAHs have been obtained. The estimated annual flux of PAHs is about 9 times greater at Industrial (193.5 mg m(-2) y(-1)) than at Marine (20.6 mg m(-2) y(-1)). These results show that study of the chemical contamination of the Venice atmosphere must take into account the PAH flux derived from marine sources as well as the continental input. PMID- 15543741 TI - Temperature dependence of the distribution of organochlorine compounds in the mosses of the Andean mountains. AB - Organochlorine compounds (OC), namely pentachlorobenzene, hexachlorobenzene, hexachlorocyclohexanes, polychlorobiphenyls (PCBs), and DDTs, have been studied in mosses distributed over three altitude gradients of the Andean mountains in Chile at 18 degrees S (3200-4500 m above sea level), 37 degrees S (345-1330 m), and 45 degrees S (10-700 m). The observed concentrations range among the lowestvalues ever reported in remote sites, but they are still higher than those found in previously studied Antarctic areas. The log transformed OC concentrations show a significant linear dependence from reciprocal of temperature independently of the origin of the compounds, e.g. industrial, agricultural, or mixed. In the case of the more volatile OC these correlations involve variance percentages higher than 50%. This good agreement gives further ground to temperature as the driving factor for the retention of long-range transported OC in remote ecosystems, including those in the southern hemisphere such as the Andean mountains. In the context of the samples selected for study, the temperature dependences in the areas of similar latitude are related to altitude. Thus, all OC in the highest altitude gradient (18 degrees S) and most compounds in the other two profiles (37 degrees S and 45 degrees S) exhibit higher concentrations with decreasing annual average temperature and thus increase with elevation above sea level. However, theoretical examination of the exponential equation relating OC concentrations to reciprocal of absolute temperatures shows that besides the temperature differences between highest and lowest elevation, the most relevant factor determining the OC concentration gradients is the lowest temperature value of each altitudinal series. That is, the point at highest elevation. PMID- 15543740 TI - Perfluoroalkyl contaminants in a food web from Lake Ontario. AB - Perfluorooctane sulfonate (PFOS) is a persistent and bioaccumulative perfluorinated acid detectable in humans and wildlife worldwide that has alerted scientists to examine the environmental fate of other fluorinated organic contaminants. Recently a homologous series of perfluoroalkyl carboxylates (PFCAs) was detected in the Arctic, yet little is known about their sources, breadth of contamination, or environmental distribution. In this study we analyzed for PFOS, the homologous series of PFCAs ranging from 8 to 15 carbons in chain length, and the PFOS-precursor heptadecafluorooctane sulfonamide (FOSA) in various organisms from a food web of Lake Ontario. The sampled organisms included a top predator fish, lake trout (Salvelinus namaycush), three forage fish species including rainbow smelt (Osmerus mordax), slimy sculpin (Cottus cognatus), and alewife (Alosa pseudoharengus), and two invertebrates Diporeia (Diporeia hoyi) and Mysis (Mysis relicta). A striking finding was that the highest mean concentration for each fluorinated contaminantwas detected in the benthic macroinvertebrate Diporeia, which occupies the lowest trophic level of all organisms analyzed. Perfluorinated acid concentrations in Diporeia were often 10-fold higher than in Mysis, a predominantly pelagic feeder, suggesting that a major source of perfluoroalkyl contaminants to this food web was the sediment, not the water. PFOS was the dominant acid in all samples, but long-chain PFCAs, ranging in length from 8 to 15 carbons, were also detected in most samples between <0.5 and 90 ng/ g. Among Mysis and the more pelagic fish species (e.g. excluding Diporeia and sculpin) there was evidence for biomagnification, but the influence of foraging on highly contaminated Diporeia and sculpin by these fish may have overestimated trophic magnification factors (TMFs), which ranged from 0.51 for FOSA to 5.88 for PFOS. By accounting for the known diet composition of lake trout, it was shown that bioaccumulation was indeed occurring at the top of the food web for all perfluoroalkyl compounds except PFOA. Future monitoring at other locations in Lake Ontario, and in other aquatic environments, is necessary to determine if these food web dynamics are widespread. Archived lake trout samples collected between 1980 and 2001 showed that mean whole body PFOS concentrations increased from 43 to 180 ng/g over this period, but not linearly, and may have been indirectly influenced by the invasion and proliferation of zebra mussels (Dreissena polymorpha) through effects on the population and ecology of forage fishes. PMID- 15543745 TI - Hydrophilic and hydrophobic sorption of organic acids by variable charge soils: effect of chemical acidity and acidic functional group. AB - Sorption of organic acids by variable-charge soil occurs through both hydrophilic and hydrophobic sorption. In this study, the effect of chemical acidity and the type of acidic functional group on the relative contribution of hydrophilic and hydrophobic processes to sorption by a gibbsite-dominated and a kaolinite dominated variable-charge soils was quantified by measuring sorption isotherms from different electrolytes (CaCl2, Ca(H2PO4)2, and KCl). The A1 soil is dominated by gibbsite whereas the DRC soil is primarily kaolinite. The organic acids investigated include five chlorinated phenols (pentachlorophenol, 2,3,4,6 tetrachlorophenol, 2,4,6-trichlorophenol, 2,4,5-trichlorophenol, and 2,4 dichlorophenol) with pKa values ranging from 4.69 to 7.85 and two acidic herbicides (2,4-D (pKa = 2.8) and prosulfuron (pKa = 3.76)) that contain carboxyl and urea functional groups, respectively. Anion exchange of chlorinated phenols and prosulfuron on both variable-charge soils as well as 2,4-D sorption on the A1 soil was linearly correlated to chemical acidity. The effective positive surface charge [AEC/(AEC + CEC)] and the anionic fraction of the organic acid in solution, which are both pH-dependent, were sufficient to estimate the contribution of anion exchange to organic acid sorption except for 2,4-D sorption by DRC soil. The latter was much greater than would be predicted from the pKa of 2,4-D. Calcium bridging between silanol edge group and 2,4-D was hypothesized and corroborated by differences in sorption measured from KCl and CaCl2 solutions. For predicting contributions from hydrophobic processes, a log-log linear relationship between pH-dependent octanol-water (Kow(pH)) and organic carbon normalized sorption coefficients (Koc(pH)) appeared adequate. PMID- 15543742 TI - Geochemical modulation of pesticide sorption on smectite clay. AB - Pesticide adsorption by soil clays can be dramatically influenced by the exchangeable cations present. Among the common exchangeable base cations in soils (Ca2+, Mg2+, K+, and Na+), K+-saturated clays frequently demonstrate the strongest affinity for pesticides. In the presence of multiple exchangeable cations in the system, we hypothesize that the magnitude of pesticide sorption to soil minerals is proportional to the fraction of clay interlayers saturated with K+ ions. To test this hypothesis, we measured sorption of three pesticides with different polarities (dichlobenil, monuron, and biphenyl) by homoionic K- and Ca smectite (SWy-2) in KCl/CaCl2 aqueous solutions. The presence of different amounts of KCl and CaCl2 resulted in varying populations of K+ and Ca2+ on the clay exchange sites. The sorption of dichlobenil and, to a lesser extent monuron, increased with the fraction of K+ on clay mineral exchange sites. Ca- and K-SWy-2 displayed the same sorption capacities for nonpolar biphenyl. X-ray diffraction patterns indicated that at lower fractions of K+-saturation, exchangeable K+ ions were randomly distributed in clay interlayers and did not enhance pesticide sorption. At higher populations of K+ (vs Ca2+), demixing occurred causing some clay interlayers, regions, or tactoids to become fully saturated by K+, manifesting greatly enhanced pesticide sorption. The forward and reverse cation exchange reactions influenced not only K+ and Ca2+ populations on clays but also the nanostructures of clay quasicrystals in aqueous solution which plays an important, if not dominant, role in controlling the extent of pesticide sorption. Modulating the cation type and composition on clay mineral surfaces through cation exchange processes provides an environmental-safe protocol to manipulate the mobility and availability of polar pesticides, which could have applications for pesticide formulation and in environmental remediation. PMID- 15543747 TI - EXAFS study of Zn sorption mechanisms on montmorillonite. AB - Extended X-ray absorption fine structure (EXAFS) analysis of zinc sorption on montmorillonite showed that different types of surface complexes or surface precipitates were formed depending on the reaction time. With an initial zinc concentration of 10(-3) M at neutral pH, zinc remained octahedrally coordinated with about six oxygen atoms at Zn-O bond distances of 2.02-2.07 A for up to six months. For samples aged up to 11 days, the Zn-Zn contribution in the second shell suggested formation of multinuclear surface complexes or surface precipitates. For samples aged 20 days and more, Zn-Zn and Zn-Si contributions in the second shell suggested formation of mixed metal coprecipitates such as a Zn phyllosilicate-like phase. Formation of these mixed metal solids probably accounts for the slow continuous sorption reaction at aging times exceeding 20 days. Sequestration of Zn in mixed metal precipitates and the stability of these phases can reduce the concentration, mobility, and toxicity of Zn in soils or sediments. PMID- 15543746 TI - Quantifying unfrozen water in frozen soil by high-field 2H NMR. AB - To understand wintertime controls of biogeochemical processes in high latitude soils it is essential to distinguish between direct temperature effects and the effects of changes in water availability mediated by freezing. Efforts to separate these controls are hampered by a lack of adequate methods to determine the proportion of unfrozen water. In this study we present a high-field 2H2O NMR method for quantifying unfrozen water content in frozen soil. The experimental material consisted of the humic layer of a boreal spruce forest soil mixed with varying proportions of quartz sand and humidified with deuterium-enriched water. The relative standard deviation of unfrozen water content (measured as NMR signal integral) was less than 2% for repeated measurements on a given sample and 3.5% among all samples, based on a total of 16 measurements. As compared to 1H NMR, this 2H NMR method was found to be superior for several reasons: it is less sensitive to field inhomogeneity and paramagnetic impurities, it gives a bigger line shape difference between the ice and liquid signal, it shows a sharper response to water fusion, and it excludes the possibility of hydrogen in the organic material interfering with the measurement. PMID- 15543744 TI - Air-water transfer of MTBE, its degradation products, and alternative fuel oxygenates: the role of temperature. AB - The gasoline oxygenate methyl tert-butyl ether (MTBE) has become one of the world's mostwidespread groundwater and surface water contaminants. As a result, there has been increasing interest in the environmental behavior of MTBE and its degradation products, mainly tert-butyl formate (TBF) and tert-butyl alcohol (TBA). In contrast, the environmental behavior of the proposed alternatives to MTBE, namely ethyl tert-butyl ether (ETBE), tert-amyl methyl ether (TAME), and diisopropyl ether (DIPE) has hardly been studied yet, although some of them are already in substantial use in various countries. A key parameter for the assessment of the fate, transport, and possible remediation of these contaminants is the air-water partitioning constant (KiH). The KiH is highly temperature dependent, and it is therefore necessary to obtain reliable experimental values at relevant temperatures. Hence, the KiH of MTBE, ETBE, TAME, and DIPE, along with the degradation products, TBF and methyl acetate, were determined from 5 degrees C-40 degrees C. The alternatives to MTBE generally had a higher KiH, which implies that, upon emission into the environment, the alternatives partition more readily into the air phase than MTBE. This may favor their use, as it is in the air phase where dilution and degradation are the most effective. The degradation products of MTBE, with the exception of TBF, have much lower KiH values at all temperatures. Hence, the degradation products will have a stronger affinity for the water phase. The temperature dependency of the kinetics of air watertransfer is discussed using a boundary layer model. Only for TBA but not for the ethers a significant effect of temperature was found. PMID- 15543749 TI - Proton binding onto soil by nonelectrostatic models: isolation and identification of mineral contributions. AB - In this paper a methodological approach is proposed to validate mechanistic modeling for proton binding onto active sites of mineral and soil samples by reducing the uncertainty and arbitrariness of model schematization. This approach is based on the quantitative formulation (X-ray calibration method) of a simulating mineral mixture (SMM) accounting for the main mineral phases in the soil (quartz, goethite, hematite, muscovite, clinochlore). Mineral and organic contributions were separated by comparing titration curves of river sediment and SMM. Specific mineral contributions to the acid properties of SMM were separated by comparing titration models of SMM and single minerals. Different nonelectrostatic models were used for titrations of SMM and single minerals: two site/three-KH models (one amphoteric plus one monoprotic site) for clay minerals and SMM; one-site/two-KH models (one amphoteric site) for goethite and hematite; and a one-site/one-KH model (one monoprotic site) for quartz. Crossed-comparisons of titration models allow for identifying and quantifying the specific contributions of the distinct edge hydroxyl groups of iron oxides, clay minerals, and quartz in the different pH ranges. In particularthe amphoteric sites of aluminosilicates mainly contribute in the acid-neutral pH range, the amphoteric sites of iron oxides take part in the neutral-basic range, and finally the monoprotic edge hydroxyl groups of quartz react in the upper basic region of pH. The good simulation of the acid-base properties of SMM (according to single mineral titration models and quantitative composition by X-ray) confirms both model schematization and SMM formulation. Speciation diagrams of the active sites of the different mineral components (aluminosilicates, iron oxides, and quartz) were obtained by implementing the database of a dedicated software with the apparent equilibrium constants regressed by titration modeling of single minerals. PMID- 15543748 TI - Thermodynamics of nitroaromatic compound adsorption from water by smectite clay. AB - Nitroaromatic compounds enter the environment through their use as explosives, pesticides, solvents, and synthetic intermediates in the manufacturing of dyes, perfumes, and drugs. Recent studies have found that many nitroaromatic compounds are strongly retained by smectites, especially K+-saturated smectites. Sorption occurs when nitroaromatic compounds replace water associated with the clay and form complexes between K+ and -NO2 groups. This study seeks to further understand nitroaromatic-clay interactions from the viewpoint of energetics. Adsorption isotherms of 1,3-dinitrobenzene, 1,4-dinitrobenzene, and 1,3,5-trinitrobenzene from aqueous solution by K+- and Ca2+-saturated smectite (SWy-2) were measured at several temperatures between 4 degrees C and 37 degrees C to determine the molar differential adsorption enthalpies. Adsorption was found to be an exothermic process on both homoionic K+- and Ca2+-smectite. The smaller adsorption enthalpy on Ca-SWy-2 was consistent with its much smaller adsorption capacity for nitroaromatics compared to K-SWy-2. Our best estimate forthe enthalpy of 1,3,5 trinitrobenzene interactions with K-SWy-2 is -124 kJ/mol, which is referenced to gas-phase 1,3,5-trinitrobenzene, corrected forthe displacement of interlayer water, and can be directly compared with quantum chemical enthalpies from the literature. Our comparable estimates for 1,3- and 1,4-dinitrobenzene interaction enthalpies are near -90 kJ/mol. We conclude that our adsorption enthalpy results are consistent with the hypothesis that nitroaromatic compounds are sorbed strongly by K-smectites because they form inner- and/or outer-sphere complexes with K+ cations in clay interlayers. Indeed, the basal spacings of rewetted clay films in the presence of nitroaromatic compounds imply that water molecules cannot effectively compete with the adsorbed nitrobenzenes for reactive sites on K-SWy-2. PMID- 15543750 TI - Cyclic exchanges and level of coupling between environmental media: intermedia feedback in multimedia fate models. AB - The importance of cyclic transport of chemicals between media in the environment can be expressed in terms of the Feedback correction factor--a multiplier that accounts for the fraction of an emission that returns to the medium of release after transfer to other media. This factor is calculated analytically by explicitly solving the appropriate system of mass balance equations or using matrix techniques. It generalizes the concept of stickiness, the ratio between the net and the overall deposition rate constants, to multipathway feedback, while providing a clearer view of the level of coupling between media and analyzing the importance of coupling. This paper first shows the usefulness of the total removal rate coefficient in each media (sum of degradation rate and all intermedia transfer rates) as a baseline to determine the chemical mass in different media, the characteristic travel distance and to understand the cyclic behavior, rather than starting from the degradation lifetimes or the overall persistence in the environment. Starting from this baseline, the importance of feedback is limited for most organic chemicals. The predicted media concentrations are influenced by less than 10% due to the cyclic nature of the intermedia transport for more than 90% of the 317 tested chemicals in a 4 compartment, steady-state, closed-system multimedia model. The Feedback correction factor is always less than a factor of 5 with the greatest values when transfer fractions are important in both directions for adjacent media. This corresponds to a restricted range in the K(AW) and K(OA) space with long chemical lifetimes in both adjacent media. This analysis of the importance of the Feedback correction factor, in conjunction with resultant criteria for when cyclic exchanges between media are likely to be significant, facilitates a more transparent understanding of how substance masses are distributed in the modeled system. It is one of the important criteria to determine to what extent media can be independently modeled. PMID- 15543752 TI - Identification of the reactive oxygen species responsible for carbon tetrachloride degradation in modified Fenton's systems. AB - The reactive oxygen species responsible for the transformation of carbon tetrachloride (tetrachloromethane, CT) by modified Fenton's reagent using hydrogen peroxide (H2O2) concentrations >0.1 M was investigated. Addition of the hydroxyl radical scavenger 2-propanol to modified Fenton's reactions did not significantly lower CT transformation rates. Scavenging by 2-propanol not only confirmed that hydroxyl radicals are not responsible for CT destruction, but also suggested that a major product of an iron (III)-driven initiation reaction, superoxide radical anion (O2-), is the species responsible for CT transformation. To investigate this hypothesis, CT degradation was studied in aqueous KO2 reactions. Minimal CT degradation was found in CT-KO2 reactions; however, when H2O2 was added to the KO2 reactions at concentrations similar to those in the modified Fenton's reactions (0.1, 0.5, and 1 M), CT degradation increased significantly. Similar results were obtained when 1 M concentrations of other solvents were added to aqueous KO2 reactions, and the observed first-order rate constant for CT degradation correlated strongly (R2 = 0.986) with the empirical solvent polarity (E(T)N) of the added solvents. The results indicate that even dilute concentrations of solvents, including H202, can increase the reactivity of O2- in water, probably by changing its solvation sphere. The higher reactivity of O2- generated in modified Fenton's reagent, which has a less polar nature due to the presence of H2O2, may result in a wider range of contaminant degradation than previously thought possible. PMID- 15543751 TI - Addition of carbon sorbents to reduce PCB and PAH bioavailability in marine sediments: physicochemical tests. AB - The addition of activated carbon as particulate sorbent to the biologically active layer of contaminated sediment is proposed as an in-situ treatment method to reduce the chemical and biological availability of hydrophobic organic contaminants (HOCs) such as polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs). We report results from physicochemical experiments that assess this concept. PCB- and PAH-contaminated sediment from Hunters Point Naval Shipyard, San Francisco Bay, CA, was contacted with coke and activated carbon for periods of 1 and 6 months. Sediment treated with 3.4 wt % activated carbon showed 92% and 84% reductions in aqueous equilibrium PCB and PAH concentrations, 77% and 83% reductions in PCB and PAH uptake by semipermeable membrane devices (SPMD), respectively, and reductions in PCB flux to overlying water in quiescent systems up to 89%. Adding coke to contaminated sediment did not significantly decrease aqueous equilibrium PCB concentrations nor PCB or PAH availability in SPMD measurements. Coke decreased PAH aqueous equilibrium concentrations by 38-64% depending on coke dose and particle size. The greater effectiveness of activated carbon as compared to coke is attributed to its much greater specific surface area and a pore structure favorable for binding contaminants. The results from the physicochemical tests suggest that adding activated carbon to contaminated field sediment reduces HOC availability to the aqueous phase. The benefit is manifested relatively quickly under optimum contact conditions and improves in effectiveness with contact time from 1 to 6 months. Activated carbon application is a potentially attractive method for in-situ, nonremoval treatment of marine sediment contaminated with HOCs. PMID- 15543757 TI - [Prediction rules in community acquired pneumonia]. AB - The determination of site of care is an essential decision in the management of patients with community-acquired pneumonia (CAP). Patients with mild to moderate CAP may be safely treated at home. Instead, those patients with severe pneumonia must be hospitalized to assure an effective treatment. Severity of CAP is associated with mortality that depends both on the patient's frailty and the intensity of lung inflammation. Because there is no single predictor factor to assess prognosis, diverse prediction rules have been developed to establish severity of CAP and guide the decision of site of care. In our country a new prediction rule, derived from hospitalized patents that incorporate simple clinical variables has been developed. However, this rule requires to be validated in the ambulatory setting before its wide spread use is suggested. Prediction rules are objective and relatively accurate models to assess prognosis that may aid clinicians to evaluate patient's risks and to improve hospitalization decisions. Nevertheless, although the prediction rules may guide the initial management of patients with CAP, they are not intended to replace the clinical judgment, which remains as the "art of medicine." PMID- 15543755 TI - Comment on "intermittent rainfall in dynamic multimedia fate modeling". PMID- 15543753 TI - Electrochemical oxidation as a final treatment of synthetic tannery wastewater. AB - Vegetable tannery wastewaters contain high concentrations of organics and other chemicals that inhibit the activity of microorganisms during biological oxidations, so biorefractory organics that are not removed by biological treatment must be eliminated by a tertiary or advanced wastewater treatment. In this paper, the applicability of electrochemical oxidation as a tertiary treatment of a vegetable tannery wastewater was investigated by performing galvanostatic electrolysis using lead dioxide (Ti/PbO2) and mixed titanium and ruthenium oxide (Ti/TiRuO2) as anodes under different experimental conditions. The experimental results showed that both the electrodes performed complete mineralization of the wastewater. In particular, the oxidation took place on the PbO2 anode by direct electron transfer and indirect oxidation mediated by active chlorine, while it occurred on the Ti/TiRuO2 anode only by indirect oxidation. Furthermore, the Ti/PbO2 gave a somewhat higher oxidation rate than that observed for the Ti/TiRuO2 anode. Although the Ti/TiRuO2 required almost the same energy consumption for complete COD removal, it was more stable and did not release toxic ions, so it was the best candidate for industrial applications. With the Ti/TiRuO2 anode, the rate of tannery wastewater oxidation increased with the current density, pH, and temperature of the solution. These results strongly indicate that electrochemical methods can be applied effectively as a final treatment of vegetable tannery wastewater allowing the complete removal of COD, tannin, and ammonium and decolorization. PMID- 15543756 TI - Comment on "sorption nonlinearity for organic contaminants with diesel soot: method development and isotherm interpretation". PMID- 15543754 TI - Degradation of endocrine disrupting chemicals bisphenol A, ethinyl estradiol, and estradiol during UV photolysis and advanced oxidation processes. AB - The degradation of three endocrine disrupting chemicals (EDCs), bisphenol A, ethinyl estradiol, and estradiol, was investigated via ultraviolet (UV) radiation photolysis and the UV/hydrogen peroxide advanced oxidation process (AOP). These EDCs have been detected at low levels in wastewaters and surface waters in both the United States and European countries, can cause adverse effects on humans and wildlife via interactions with the endocrine system, and thus must be treated before entering the public drinking water supply. Because many EDCs can only be partially removed with conventional water treatment systems, there is a need to evaluate alternative treatment processes. For each EDC tested, direct UV photolysis quantum yields were derived for use with both monochromatic low pressure (LP) UV lamps and polychromatic medium-pressure (MP) UV lamps and second order hydroxyl radical rate constants were developed. These parameters were utilized to successfully model UV treatment of the EDCs in laboratory and natural waters. The polychromatic MP UV radiation source was more effective for direct photolysis degradation as compared to conventional LP UV lamps emitting monochromatic UV 254 nm radiation. However, in all cases the EDCs were more effectively degraded utilizing UV/H2O2 advanced oxidation as compared to direct UV photolysis treatment. PMID- 15543759 TI - [Development of an instrument to assess the severity of community acquired pneumonia among hospitalized patients]. AB - BACKGROUND: Community acquired pneumonia (CAP) severity assessment is crucial. AIM: To develop a practical clinical severity assessment model for stratifying immunocompetent adult patients hospitalized with CAP into different management groups. PATIENTS AND METHODS: During a 24 months period, 455 adult patients (250 male, mean age 69 +/- 19 years old) were evaluated. All the relevant clinical information recorded and they were followed during hospital stay until discharge or death. Mortality until 30 days after admission was determined. RESULTS: The mean hospital length of stay was 9.9 +/- 9.4 days and 76% had an underlying disease. In hospital mortality was 7.6% and 10.1% at 30 days follow up. Admission prognostic factors associated with high mortality at 30 days follow up were: advanced age, presence of comorbidity, suspicion of aspiration, duration of symptoms < or = 2 days, altered mental status, absence of cough, fever and cbills, low blood pressure, tachypnea, hypoxemia and multilobar radiographic pulmonary infiltrates. A clinical prognostic index derived from a logistic regression analysis including five independent variables associated with mortality (confuson, comorbidity, low systolic blood pressure, temperature < 37.5 degrees C and respiratory rate > 20/min), enabled patients to be stratified according to increasing risk of mortality: class 1: 0.9%, class 2: 4.9%, class 3: 14.2%, and class 4: 35.6%. CONCLUSION: A simple clinical severity assessment tool based on confusion, comorbidity, blood pressure, temperature and respiratory rate could be used to stratify patients with CAP into different risk class categories and management groups. PMID- 15543760 TI - [Presence of DRD4/7R and DAT1/10R allele in Chilean family members with attention deficit hyperactivity disorder]. AB - BACKGROUND: Genes for dopamine receptor DRD4 and dopamine transporter DAT1 have been implicated in attention deficit with hyperactivity disorder (ADHD). However, the findings are not conclusive. More studies in populations with different genetic backgrounds may contribute to solve the discrepancies observed. AIM: To test the hypothesis that affected members of Chilean families exhibit higher frequencies of the DRD4/7R and DAT1/10R alleles then their healthy sibs. MATERIAL AND METHODS: The parents of 51 children belonging to families of the Metropolitan Region of Chile, were approached to obtain clinical histories and blood samples, after the signature of a written informed consent. ADHD was diagnosed according to DSM-IV criteria, ancd intellectual coefficient was tested using the WISC-R test. Genomic DNA was extracted from lymphocytes and amplified by PCR. RESULTS: The 7R allele was identified in 13 out of 26 subjects diagnosed as ADHD and in 6 of 25 healthy sibs (p < 0.05). Parents with a history of ADHD, were conmpared with their healthy counterparts, exhibiting an identical tendency, that did not reach statistical significance. No significant differences in the frequencies of DAT1/10R alleles, were observed between cases and controls or their parents. CONCLUSIONS: Our results showed that ADHD in Chilean families is associated with the presence of DRD4/7R allele. PMID- 15543762 TI - [Polymorphisms of the multiple drug resistance gene (MDR1) in Mapuche, Mestizo and Maori populations in Chile]. AB - BACKGROUND: There are significant differences in drug responses among different ethnic groups. The multidrug transporter P-gp, encoded by the MDR1 gene, plays a key role in determining drug bioavailability, and an association between a polymorphism in exon 26 (C3435T) and lower P-gp expression has been found. The co segregation of this polymorphism with the polymorphism in exon 12 (C1236T) and in exon 21 (G2677T/A) determines several MDR1 haplotypes in humans. AIM: To characterize the polymorphisms of exons 26, 21 and 12 of the MDR1 gene in different Chilean populations. MATERIAL AND METHODS: Using a polymerase chain reaction and restriction fragment length polymorphism technique, we studied the allelic frequencies and the distribution of MDR1 haplotypes in 3 Chilean populations: Mestizo (n=104), Mapuche (n=96, living in the National Reservation of the Huapi Island, Ranico Lake) and Maori (n=52, living in Eastern Island). RESULTS: The frequency of the normal MDR1*1 haplotype, without mutations, was lower in Mapuches than in Mestizos or Maoris (p<0.005) but similar to that reported in Asian population (p=0.739), probably due to the Asian origin of the Amerindian populations. In addition, the MDR1*l haplotype fequency hin Mestizos was similar to the frequency reported in Caucasians (p=0.49), in agreement with the origin of our population, with a strong influence of Caucasian genes from the Spanish conquerors. The MDR1*2 haplotype distribution, with the three polymoyphisms and probably lower multidrug transporter expression, was similar in the three Chilean populations studied (p>0.0.5), but lower than the frequencies reported in Caucasians or Asians (p<0.05). CONCLUSIONS: We found significant differences in the frequencies of genetic polymorphisms of the MDR1 gene in Chilean populations, related to the ethnic origins of our ancestors. PMID- 15543758 TI - [Prognostic value of hyperuricemia in chronic heart failure]. AB - BACKGROUND: Hyperuricemia has been proposed as a risk marker in chronic heart failure, but its value as an independent prognostic is not well established. AIM: To determine the prognostic value of hyperuricemia, in patients with chronic stable heart failure. PATIENTS AND METHODS: Forty six male patients with chronic heart failure, aged 62 +/- 13 years, were studied. Their election fraction was less than 40% and their serum creatinine was less than 2 mg/dl. Serum uric acid and catecholamines, maximal oxygen consumption (VO2 max) and left ventricular ejection fraction were measured. Mortality and the need for cardiac transplant were recorded as endpoints during a mean follow up of 39 +/- 18 months. The relationship between basal measures and the occurrence of events was analyzed using univariate and multivariate methods. RESULTS: Basal VO2 max and left ventricular ejection fraction were 16 +/- 4.6 ml/kg/min and 22 +/- 7% respectively. Eighteen patients died and three required transplantation during the follow up. Patients reaching these endpoints had a lower VO2 max and left ventricular ejection fraction and higher uric acid levels. Multivariate analysis accepted left ventricular ejection fraction (relative risk 0.89, 95% CI 0.82 0.97) and serum uric acid (relative risk 1.335 95% CI 1.02-1.74) as significant predictors of events. The relative risk for cardiac transplantation was 7.07 times higher among those with a serum uric acid over 7 mg/dl. CONCLUSIONS: A high serum uric acid is an independent predictor of bad prognosis in patients with stable chronic heart failure. PMID- 15543761 TI - [Free urinary cortisol is elevated in patients with low-renin essential hypertension]. AB - BACKGROUND: Glucocorticoids play a key role in blood pressure (BP) control and are associated with hypertension in patients with Cushing's syndrome. A number of reports indicate that cortisol (F) may be involved in etiology of essential hypertension (EH). F can bind to the mineralocorticoid receptor, triggering both sodium and water reabsorption in kidney, increase BP and cause renin suppression. AIM: To evaluate urinary free cortisol (UFF) excretion as a potential intermediate phenotype of essential hypertension and correlate F level with plasma renin activity (PRA) and serum aldosterone (SA). PATIENTS AND METHODS: We recruited 132 EH patients and 16 normotensive healthy controls. Blood samples and 24 hours urine were collected for PRA, SA and UFF analysis. Differences in UFF excretion between sexes were normalized by urinary creatinine (Creat) excretion. The upper limit of UFF/Creat was determined in normotensives considering the mean value plus 2 standard deviations. According to this value, subjects were classified as having high or normal UFF. RESULTS: In EH patients and in normotensives, the UFF/Creat was 36.9 +/- 17.0 microg/gr and 30.9 +/- 8.8 microg/gr, respectively. The upper limit was set at 48.5 microg/gr. A high UFF/Creat was found in 20/132 EH (15%) patients and 0/16 normotensive subjects. EH patients with high UFF showed lower PRA levels than patients with normal cortisol levels (0.78 +/- 0.47 vs. 1.13 +/- 0.66 ng/ml x h, respectively, p=0.027) and lower SA values (4.52 +/- 1.65 vs 6.34 +/- 3.37 ng/dl, respectively, p=0.018). There was a negative correlation between UFF and PRA (r=-0.176, p=0.044) and between UFF and SA (r=-0.183, p=0.036). CONCLUSIONS: We have identified a subgroup of EH patients with increased UFF excretion. Patients with the highest UFF showed lower renin and aldosterone levels. These data suggest a potential influence of cortisol in the genesis of hypertension. PMID- 15543765 TI - Non-urgent patients in an Emergency Medical Service. AB - BACKGROUND: Emergency Medical Services (ES) are medical-surgical facilities where extremely ill patients, victims of accidents, intoxications and other urgent clinical situations should be rapidly and efficiently attended. However, a worldwide experience is that a large proportion of patients who require attention in an ES do not qualify as emergencies and they obstruct the possibilities of other patients that should be attended immediately. AIM: To evaluate the medical records in our ES and to assess whether patients understand what is a true emergency and which is the role of an ES. METHODS: Patients attended in the ES of Haydarpasa Military Teaching Hospital in 2001-2002 were classified according to their diagnosis. A random sample of patients and relatives were surveyed about why they required medical attention in the ES. RESULTS: 77,134 patients (39,327 female, 51%) were attended. The most common illness diagnosed was upper respiratory tract infection (18,423 patients, 23.9%). The true emergency condition rate in patients presenting to our ES was lower (44%) than the non emergency condition rate. CONCLUSIONS: Improvements in public education are needed to solve this problem. Also, Primary Care facilities should be improved in order to reduce the demand for medical attention in ES by non-urgent patients. PMID- 15543768 TI - [Work absenteeism in Chile according to the type of work place]. AB - BACKGROUND: The issue of medically justified work absenteeism has a great relevance in Chile at the present moment. AIM: To analyze sick leaves among people working in hospitals, mines, automotive industry and universities. MATERIAL AND METHODS: Analysis of 14 thesis and research papers about absenteeism in Chile. The incapacity rate (number of days with sick leave per worker per year, the frequency rate (number of sick leaves per year per worker) and the severity rate (mean duration of sick leaves) were calculated. The diseases causing the highest rates of absenteeism were also recorded. RESULTS: The mean age of the studied populations was 36 years old and the most common diseases causing absenteeism were respiratory, rheumatologic and trauma. Hospital workers had the highest incapacity rate with 14.3 days of sick leave per worker per year, followed by mining industry with 12 days, automotive industry with 7.1 days and universities with 6 days. CONCLUSIONS: In Chile, respiratory diseases are the main cause of sick leaves and hospital workers have the highest incapacity rate. PMID- 15543763 TI - [Promoter methylation profile in breast cancer]. AB - BACKGROUND: Genomic DNA methylation, mutations and allelic deletions explain the inactivation of genes involved in cell proliferation and cell cycle control mechanisms. AIM: To analyze the methylation pattern of important genes related to different carcinogenic mechanisms in patients with breast cancer and the relationship with its biological behavior. MATERIAL AND METHODS: Seventy fresh frozen breast cancer samples were selected. The methylation specific PCR (MSP) test was used to analyze promoter methylation status for genes CDKN2A (p16), hMLH1, APC, CDH1 (Cadherin E) and FHIT. RESULTS: We found methylation in at least one of the genes studied in 88% of cases and in 3 or more genes in 40.5% of cases. The frequencies of promoter hypermethylation of CDKN2A, hMLH1, APC, CDH1 and FHT were 41.4%, 11.4%, 52.9%, 70% and 42.9%, respectively. We found a relationship between CDKN2A methylatlon and better survival (p=0.002). CDH1 methylation and poor histological differentiation (p=0.007), hMLH1 methylation and non-Mapuche ethnicity (p=-0.03), APC methylation and larger tumor size (p<0.05), FHIT methylatton and lack of estrogen rectptor IHC expression (p<0.05). CONCLUSIONS: The high frequency of promoter methylation in patients with breast cancer confirms its role in breast carcinogenesis. The finding of alterations in the methylation pattern of genes studied and its association with prognostic factors is a helpful tool in the search of new criteria for clinical and therapeutic decision making. PMID- 15543764 TI - [Laboratory diagnosis of patients with exanthematic or febrile syndromes occurring in the Zulia State, Venezuela, during 1998]. AB - BACKGROUND: Exanthematic diseases are a group of syndromes mainly caused by acute viral infections. AIM: To obtain information about the viruses that cause exanthematic diseases in our region. PATIENTS AND METHODS: During 1998, 267 serum samples from patients with an acute rash or patients presenting a febrile syndrome accompanied by enlarged lymph nodes, headache and other symptoms, were collected. Specific antibody of the IgM class (anti-IgM) against Rubella, Measles, Dengue types 1-4 and Cytomegalovirus (CMV) were measured by immunoenzymatic assay (EIA). Epstein-Barr virus (EBV) antibodies were measured by immunofluorescence. RESULTS: An etiologic agent was detected in 208 cases (77.9%). Cases due to Dengue (40.6%) and Rubella (21%) viruses predominated, but the frequency of other agents was also high in specific age groups. The agreement between the clinical suspicion and the laboratory findings varied broadly, from a 100% for suspected Dengue to just a 14.8% for cases of suspected CMV infection. CONCLUSIONS: Dengue was the most common viral exanthematic disease in the Zulia State during 1998. PMID- 15543767 TI - [Familial generalized thyroid hormone resistance. Report of one case]. AB - Thyroid hormone resistance is a rare autosomal dominant disease associated, in more than 90% of cases, to mutations in the beta thyroid hormone receptor. We report a 23 years old male that consulted for a psychiatric condition. Clinically, the patient was euthyroid in spite of high total and free T4 and T3 concentrations, while TSH remained normal. Also, TSH showed a five fold increase under TRH stimulation. The mother and one of his brothers had the same pattern of abnormal serum thyroid hormones. We discuss the diagnostic considerations and the protocol to study this rare pathology. PMID- 15543766 TI - [Hepatic veno-occlusive disease, idiopathic ulcerative colitis and portal thrombosis. Report of one case]. AB - We report a previously healthy 29 years old man, presenting with a sudden episode of abdominal pain, mild jaundice, hepatomegaly and ascites. Magnetic resonance imaging study and liver biopsy were compatible with veno-occlusive disease. Incidentally, an ulcerative colitis and portal vein thrombosis were diagnosed. Anticoagulant treatment was started, with good clinical and radiological response. Veno-occlusive disease of the liver must be suspected In cases of liver failure and ascites associated to procoagulant conditions. PMID- 15543769 TI - [Intercultural health care policy from the perspective of health care providers and Mapuche clients]. AB - BACKGROUND: Intercultural health is becoming an emergent topic in the design of health care programs for Mapuche people of Chile. This process faces important challenges such as the scarce theoretical support about the meaning of intercultural health and their practical consequences for providers and clients. AIM: To explore the perception in providers and Mapuche clients about intercultural health. MATERIAL AND METHODS: A survey performed in 11 counties with the highest concentration of Mapuche people, of the IX region of Chile. The perception about the development of a new health policy specially designed for Mapuche patients was surveyed in 399 Mapuche patients and 64 providers of primary health care centers. RESULTS: Mapuche clients considered, as the main regional challenges, the indifference and discrimination of health care teams towards Mapuche patients, aggravated by the indifference of authorities. Providers considered that the main problem was a lack of knowledge about Mapuche culture and skills to deal with this ethnic group. Patients and providers agreed on the need to use Mapuche dialect in health care attentions, to coordinate actions with traditional healers and to accept ethnical therapeutic practices. CONCLUSIONS: There is scarce agreement between providers and Mapuche clients about the need for an special intercultural health policy, its contents, and the regional conditions for its implementation and development. PMID- 15543770 TI - [Advances in cellular immunotherapy for malignant melanoma]. AB - An alternative strategy for cancer treatment is the manipulation of the immune system, denominated cancer immunotherapy. The immunotherapeutical use of cells of the immune system, like dendritic cells (DC), is being explored in different clinical protocols. Recently, we finalized a clinical phase I protocol, for the treatment of malignant melanoma, using DCs loaded with tumor lysates. Our results indicate that the subcutaneous application of DCs do not produce adverse effects. We also observed an increase of tumor specific T lymphocytes precursors in the blood, associated to hypersensitivity reactions (DTH) in 60% of the treated patients. In most cases, an stability in the disease was observed, although without a significant association between vaccination and survival. Additionally, therapies based on Interleukin-2 (IL-2) have been used with relative success in the treatment of some kind of tumors since 1985. However, problems associated to the toxicity of IL-2 still restrict its massive use. Our direct experience with the use of IL-2, indicates that low doses and its subcutaneous application, maintains the beneficial effects for patients, eliminating the adverse effects. Based on the accumulated evidence during last the five years, we decided to implement an optimized clinical protocol, which alternatively combines dendritic cells vaccines with the use of low doses of IL-2 for the reinforcement of the immunological system. PMID- 15543771 TI - [The influence of phychological features and learning styles on the academic performance of medical students]. AB - BACKGROUND: The degree of difficulty we experience while learning different concepts and skills depends, among other things, on our psychological features and learning style. This may be particularly true for medical students, whose formation involves the acquisition of multiple cognitive, affective and psychomotor skills. AIM: To assess whether the psychological features and learning styles of medical students are associated with their academic performance. SUBJECTS AND METHODS: The psychological preferences and learning styles of 66 students of the 2001-graduating cohort were determined with the Myers Briggs Type Inventory (MBTI) and the Kolb Learning Style Inventory (LSI), respectively. The academic performance was assessed by the Calificacion Medica Nacional (CMN), Chile and by the marks obtained during the Basic (1st to 3rd), Preclinical (4th and 5th) and Clinical (6th and 7th) years of undergraduate training. RESULTS: The psychological features, together with the sex of students were found to be associated with the performance in the Preclinical and Clinical years, and to the CMN. In men, the interest and ability to communicate with people and the concern for harmony, and in women the tendency to function in a systematic and orderly way are the features associated to high academic performance. No associations were found between learning styles and academic performance. CONCLUSIONS: The finding that the psychological preferences of medical students are relevent to their academic performance opens a new perspective to analyze the medical education and to design programs aimed at improving learning. PMID- 15543772 TI - [Blinding for clinical trials. Does it matter?]. PMID- 15543774 TI - [Training in hematology, in Chile]. PMID- 15543773 TI - [Critical appraisal: Subcutaneous adjusted-dose unfractionated heparin vs fixed dose low-molecular-weight heparin in the initial treatment of venous thromboembolism. Prandoni P, Carnovali M, Marchiori A, Galilei investigators. Arch Intern Med 2004; 164: 1077-83]. AB - BACKGROUND: Few reports have addressed the value of unfractionated heparin (UFH) or low-molecular-weight heparin in treating the fill spectrum of patients with venous thromboembolism (VTE), including recurrent VTE and pulmonary embolism. METHODS: In an open, multicenter clinical trial, 720 consecutive patients with acute symptomatic VTE, including 119 noncritically ill patients (16.5%) with pulmonary embolism and 102 (14.2%) with recurrent VTE, were randomly assigned to treatment with subcutaneous UFH with dose adjusted by activated partial thromboplastin time by means of a weight-based algorithm (preceded by an intravenous loading dose), or fixed-dose (adjusted only to body weight) subcutaneous nadroparin calcium. Oral anticoagulant therapy was started concomitantly and continued for at least 3 months. We recorded the incidence of major bleeding during the initial heparin treatment and that of recurrent VTE and death during 3 months of follow-up. RESULTS: Fifteen (4.2%) of the 360 patients assigned to UFH had recurrent thromboembolic events, as compared with 14 (3.9%) of the 360 patients assigned to nadroparin (absolute difference between rates, 0.3%; 95% confidence interval, -2.5% to 3.1%). Four patients assigned to UFH (1.1%) and 3 patients assigned to nadroparin (0.8%) had episodes of major bleeding (absolute difference between rates, 0.3%; 95% confidence interval, -1.2% to 1.7%). Overall mortality was 3.3% in each group. CONCLUSIONS: Subcutaneous UFH with dose adjusted by activated partial thromboplastin time by means of a weight based algorithm is as effective and safe as fixed-dose nadroparin for the initial treatment of patients with VTE, including those with pulmonary embolism and recurrent VTE. PMID- 15543775 TI - Educational programs for imaging physicists should emphasize the science of imaging rather than the technology of imaging. PMID- 15543776 TI - Educational programs for imaging physicists should emphasize the science of imaging rather than the technology of imaging. PMID- 15543777 TI - Comparison of small photon beams measured using radiochromic and silver-halide films in solid water phantoms. AB - In this study, we compared the dosimetric properties of four of the most commonly used films for megavoltage photon-beam dosimetry when irradiated under identical conditions by small multileaf-collimator (MLC) defined beamlets. Two silver halide films (SHFs), Kodak XV2 and EDR2, and two radiochromic films (RCFs), Gafchromic HS and MD55-2, were irradiated by MLC-defined 1 x 1 cm2 beamlets from a Varian 2100 C/D linac equipped with a 120-leaf MLC. The beamlets were delivered with the accelerator gantry set laterally (90 degrees rotation) upon a solid water compression film phantom at 100 cm source-to-surface distance which was positioned with the films parallel to the beam axis. Beamlets were delivered at central axis, 5.0 cm, and 10.5 cm off-axis for both leaf-end and leaf-side defined beamlets. The film dosimetry was performed using a quantitative optical density (OD) imaging system that was validated in a previous study. No significant differences between SHF and RCF measurements were observed in percentage depth doses, horizontal depth profiles, or two-dimension spatial isodose distributions in both the central axis and off-axis measurements. We found that regardless of the type of film used, RCF or SHF, a consistent data set for small beam dose modeling was generated. Previous validation studies based on the use of RCF and OD imaging system would indicate that all film produce an accurate result for small beam characterization. PMID- 15543778 TI - Respiration tracking in radiosurgery. AB - Respiratory motion is difficult to compensate for with conventional radiotherapy systems. An accurate tracking method for following the motion of the tumor is of considerable clinical relevance. We investigate methods to compensate for respiratory motion using robotic radiosurgery. In this system the therapeutic beam is moved by a robotic arm, and follows the moving target through a combination of infrared tracking and synchronized x-ray imaging. Infrared emitters are used to record the motion of the patient's skin surface. The position of internal gold fiducials is computed repeatedly during treatment, via x-ray image processing. We correlate the motion between external and internal markers. From this correlation model we infer the placement of the internal target during time intervals where no x-ray images are taken. Fifteen patients with lung tumors have recently been treated with a fully integrated system implementing this new method. The clinical trials confirm our hypothesis that internal motion and external motion are indeed correlated. In a preliminar study we have extended our work to tracking without implanted fiducials, based on algorithms for computing deformation motions and digitally reconstructed radiographs. PMID- 15543779 TI - DMLC leaf-pair optimal control of IMRT delivery for a moving rigid target. AB - In dynamic multileaf collimation (DMLC), pairs of servo-controlled leaves sweep across the target to deliver the modulated radiation intensity map while the beam is on continuously. The mathematical model for dynamic, optimal control of a single leaf pair has been developed for the case of a rigid target, translating parallel to the leaf trajectories. This mathematical model ensures delivery of the modulated intensity map while minimizing beam-on time. Numerical solutions of the model are presented here for optimal IMRT delivery for stationary and oscillating targets, together with a discussion of the results. Comparisons between solutions for stationary and mobile targets, as well as comparisons between optimal and suboptimal algorithms, are provided. These comparisons allow us to estimate potential gains in the effectiveness of DMLC IMRT delivery when it is based on optimal algorithms. PMID- 15543780 TI - Nitroxide conjugate of a thermally responsive elastin-like polypeptide for noninvasive thermometry. AB - Hyperthermia, as an adjuvant with radiation and chemotherapy, has shown promise in the treatment of cancer. The relevant biological effects of a hyperthermia treatment are both time and temperature-dependent, creating a need for accurate thermometry. We present a novel noninvasive thermometry modality that combines a temperature responsive biopolymer, the elastin-like polypeptide (ELP), and nitroxide to produce an ELP-nitroxide conjugate. When examined with electron paramagnetic resonance (EPR) spectroscopy, the ELP-nitroxide conjugate has temperature-dependent spectral line widths whose predictive accuracy is approximately 0.3 degrees C (80 microM). We believe that the temperature dependent changes observed in the EPR spectrum are due to the combined effect of temperature, viscosity and effective radius on the rotational correlation time of the ELP-nitroxide conjugate. PMID- 15543781 TI - Direct analysis of molybdenum target generated x-ray spectra with a portable device. AB - In routine applications, information about the photon flux of x-ray tubes is obtained from exposure measurements and cataloged spectra. This approach relies mainly on the assumption that the real spectrum is correctly approximated by the cataloged one, once the main characteristics of the tube such as voltage, target material, anode angle, and filters are taken account of. In practice, all this information is not always available. Moreover, x-ray tubes with the same characteristics may have different spectra. We describe an apparatus that should be useful for quality control in hospitals and for characterizing new radiographic systems. The apparatus analyzes the spectrum generated by an x-ray mammographic unit. It is based on a commercial CZT produced by AMPTEK Inc. and a set of tungsten collimator disks. The electronics of the CZT are modified so as to obtain a faster response. The signal is digitized using an analog to digital converter with a sampling frequency of up to 20 MHz. The whole signal produced by the x-ray tube is acquired and analyzed off-line in order to accurately recognize pile-up events and reconstruct the emitted spectrum. The energy resolution has been determined using a calibrated x-ray source. Spectra were validated by comparison of the HVL measured using an ionization chamber. PMID- 15543782 TI - Dosimetric evaluation of the clinical implementation of the first commercial IMRT Monte Carlo treatment planning system at 6 MV. AB - In this work we dosimetrically evaluated the clinical implementation of a commercial Monte Carlo treatment planning software (PEREGRINE, North American Scientific, Cranberry Township, PA) intended for quality assurance (QA) of intensity modulated radiation therapy treatment plans. Dose profiles calculated in homogeneous and heterogeneous phantoms using this system were compared to both measurements and simulations using the EGSnrc Monte Carlo code for the 6 MV beam of a Varian CL21EX linear accelerator. For simple jaw-defined fields, calculations agree within 2% of the dose at d(max) with measurements in homogeneous phantoms with the exception of the buildup region where the calculations overestimate the dose by up to 8%. In heterogeneous lung and bone phantoms the agreement is within 3%, on average, up to 5% for a 1 x 1 cm2 field. We tested two consecutive implementations of the MLC model. After matching the calculated and measured MLC leakage, simulations of static and dynamic MLC defined fields using the most recent MLC model agreed to within 2% with measurements. PMID- 15543783 TI - An algorithm for independent verification of Gamma Knife treatment plans. AB - A formalism for independent treatment verification has been developed for Gamma Knife radiosurgery in analogy to the second checks being performed routinely in the field of external beam radiotherapy. A verification algorithm is presented, and evaluated based on its agreement with treatment planning calculations for the first 40 Canadian Gamma Knife patients. The algorithm is used to calculate the irradiation time for each shot, and the value of the dose at the maximum dose point in each calculation matrix. Data entry consists of information included on the plan printout, and can be streamlined by using an optional plan import feature. Calculated shot times differed from those generated by the treatment planning software by an average of 0.3%, with a standard deviation of 1.4%. The agreement of dose maxima was comparable with an average of -0.2% and a standard deviation of 1.3%. Consistently accurate comparisons were observed for centrally located lesions treated with a small number of shots. Large discrepancies were almost all associated with dose plans utilizing a large number of collimator plugs, for which the simplifying approximations used by the program are known to break down. PMID- 15543784 TI - Reproducibility of multi-slice spiral computed tomography scans: an experimental study. AB - In multi-slice spiral computed tomography (CT) images interpolation artifacts are present. The relationship between the x-ray tube rotation angle and these artifacts is demonstrated. A head phantom was repeatedly scanned with a four slice CT scanner at different pitch values. Two scans, made with identical scan parameters, nearly always have different x-ray tube starting angles. Consequently, artifacts appeared differently and residual artifacts appeared when subtractions were made. We conclude that only if the x-ray tube starting angle is equal for both scans or if a very low pitch is used, images are highly reproducible. PMID- 15543785 TI - Dose enhancement close to platinum implants for the 4, 6, and 10 MV stereotactic radiosurgery. AB - Three photon interaction processes, namely, the photoelectric effect, Compton effect, and pair production, can occur when materials with high atomic numbers are irradiated by the high- and low-energy bremsstrahlung photons from a linear accelerator. A dose enhancement, due to the photoelectric effect and pair production, near targets with platinum implants (with a high atomic number) in radiosurgery cannot be predicted by the XKnife radiosurgery treatment planning system. In the present work, Monte Carlo simulations using PRESTA EGS4 were employed to investigate the resulting dose enhancements from 4, 6, and 10 MV energies commonly used in the stereotactic radiosurgery system. Dose enhancements from 32% to 68% were observed close to the platinum implant for the above energies when using a 12.5 mm collimator. Comparatively higher dose enhancements were observed when using smaller collimators. It was found that this dose enhancement increased with beam energy but decreased as beam size increased. PMID- 15543786 TI - The behavior of several microionization chambers in small intensity modulated radiotherapy fields. AB - The aim of this work is to compare different ion chambers available for dose measurements in small fields used in intensity modulated radiotherapy. Some dosimetric aspects, related to these small radiation fields, i.e., lack of electronic lateral equilibrium and steep dose region, must be evaluated, in order to obtain an accurate technique implementation. Furthermore, the size of the sensitive volume of the chambers compared with the mapping of the beams or segments needs consideration. If the size of the chamber is too large for the flatness of the field, the measurement can deviate from the expected absorbed dose at a point. We propose a comparison of various dosimetric values between different microionization chambers with respect to a smaller dosimeter, such as the diamond detector. PMID- 15543787 TI - Steepest changes of a probability-based cost function for delineation of mammographic masses: a validation study. AB - Our purpose in this work was to develop an automatic boundary detection method for mammographic masses and to rigorously test this method via statistical analysis. The segmentation method utilized a steepest change analysis technique for determining the mass boundaries based on a composed probability density cost function. Previous investigators have shown that this function can be utilized to determine the border of the mass body. We have further analyzed this method and have discovered that the steepest changes in this function can produce mass delineations that include extended projections. The method was tested on 124 digitized mammograms selected from the University of South Florida's Digital Database for Screening Mammography (DDSM). The segmentation results were validated using overlap, accuracy, sensitivity, and specificity statistics, where the gold standards were manual traces provided by two expert radiologists. We have concluded that the best intensity threshold corresponds to a particular steepest change location within the composed probability density function. We also found that our results are more closely correlated with one expert than with the second expert. These findings were verified via Analysis of Variance (ANOVA) testing. The ANOVA tests obtained p-values ranging from 1.03 x 10(-2)-7.51 x 10( 17) for the single observer studies and 2.03 x 10(-2)-9.43 x 10(-4) for the two observer studies. Results were categorized using three significance levels, i.e., p<0.001 (extremely significant), p <0.01 (very significant), and p <0.05 (significant), respectively. PMID- 15543788 TI - Adaptation of GEANT4 to Monte Carlo dose calculations based on CT data. AB - The GEANT4 Monte Carlo code provides many powerful functions for conducting particle transport simulations with great reliability and flexibility. However, as a general purpose Monte Carlo code, not all the functions were specifically designed and fully optimized for applications in radiation therapy. One of the primary issues is the computational efficiency, which is especially critical when patient CT data have to be imported into the simulation model. In this paper we summarize the relevant aspects of the GEANT4 tracking and geometry algorithms and introduce our work on using the code to conduct dose calculations based on CT data. The emphasis is focused on modifications of the GEANT4 source code to meet the requirements for fast dose calculations. The major features include a quick voxel search algorithm, fast volume optimization, and the dynamic assignment of material density. These features are ready to be used for tracking the primary types of particles employed in radiation therapy such as photons, electrons, and heavy charged particles. Recalculation of a proton therapy treatment plan generated by a commercial treatment planning program for a paranasal sinus case is presented as an example. PMID- 15543789 TI - Influence of x-ray pulse parameters on the image quality for moving objects in digital cardiac imaging. AB - The image quality of a single frame in a modern cardiac imaging x-ray facility can be improved by adjusting the automatic pulse exposure parameters. The effects of acquisition rate on patient dose and the detectability of moving objects have been fully described in scientific literature. However, the influence of automatic pulse exposure parameters is still to be determined. Images of a moving wheel (with lead wires) were acquired using an H5000 Philips Integris cardiac x ray system. Poly(methylmethacrylate) plastic samples 20 and 30 cm thick were employed as the build-up phantom to simulate a patient. The images were obtained using preset clinical parameters for cardiac imaging procedures. The signal detectability and motion blur of a contrast bar at a transversal speed in the range of 100-150 mm/s were evaluated with a cine pulse width of 3, 5, 7, and 10 ms under automatic mA kV regulation. Two levels of exposure at the image intensifier entrance were included in this study. Signal detectability was analyzed in terms of the signal-to-noise ratio (SNR) and the value of SNR2/entrance surface dose. The blurring was modeled as a Gaussian-shaped blurring function, and the motion blur was expressed in terms of the peak full width at half maximum and amplitude (apparent contrast) of the resolution functions. A contrast bar simulating a vessel in motion at the maximum velocities of typical cardiac structures was exposed. Severe loss of image quality occurred at pulse widths > or =7 ms. It is also shown that below 5 ms static nonlinearities, likely caused by the need to use a large focus for cine acquisition, dominate the blurring process. PMID- 15543790 TI - Determination of the reference air kerma rate for 192Ir brachytherapy sources and the related uncertainty. AB - Different methods exist to determine the air kerma calibration factor of an ionization chamber for the spectrum of a 192Ir high-dose-rate (HDR) or pulsed dose-rate (PDR) source. An analysis of two methods to obtain such a calibration factor was performed: (i) the method recommended by [Goetsch et al., Med. Phys. 18, 462-467 (1991)] and (ii) the method employed by the Dutch national standards institute NMi [Petersen et al., Report S-EI-94.01 (NMi, Delft, The Netherlands, 1994)]. This analysis showed a systematic difference on the order of 1% in the determination of the strength of 192Ir HDR and PDR sources depending on the method used for determining the air kerma calibration factor. The definitive significance of the difference between these methods can only be addressed after performing an accurate analysis of the associated uncertainties. For an NE 2561 (or equivalent) ionization chamber and an in-air jig, a typical uncertainty budget of 0.94% was found with the NMi method. The largest contribution in the type-B uncertainty is the uncertainty in the air kerma calibration factor for isotope i, N(i)k, as determined by the primary or secondary standards laboratories. This uncertainty is dominated by the uncertainties in the physical constants for the average mass-energy absorption coefficient ratio and the stopping power ratios. This means that it is not foreseeable that the standards laboratories can decrease the uncertainty in the air kerma calibration factors for ionization chambers in the short term. When the results of the determination of the 192Ir reference air kerma rates in, e.g., different institutes are compared, the uncertainties in the physical constants are the same. To compare the applied techniques, the ratio of the results can be judged by leaving out the uncertainties due to these physical constants. In that case an uncertainty budget of 0.40% (coverage factor=2) should be taken into account. Due to the differences in approach between the method used by NMi and the method recommended by Goetsch et al., an extra type-B uncertainty of 0.9% (k= 1) has to be taken into account when the method of Goetsch et al. is applied. Compared to the uncertainty of 1% (k= 2) found for the air calibration of 192Ir, the difference of 0.9% found is significant. PMID- 15543791 TI - Incomplete skin representation in digital mammograms. AB - Flat-panel digital detector systems have limited dynamic range and saturate at a particular x-ray exposure. Hence some of the breast edge may not be represented in the displayed image. We developed a model to estimate the amount of skin loss. Model predictions agreed well with phantom measurements. In our database of 884 clinical digital mammograms, 98% had saturated backgrounds. The estimated skin loss exceeded 0.5 mm in 5% of images and 1.0 mm in 0.7% of images. Any skin thickening that is present should still be visualized, so we conclude that any skin-line loss may not be of clinical significance. PMID- 15543792 TI - Inverse treatment planning with adaptively evolving voxel-dependent penalty scheme. AB - In current inverse planning algorithms it is common to treat all voxels within a target or sensitive structure equally and use structure specific prescriptions and weighting factors as system parameters. In reality, the voxels within a structure are not identical in complying with their dosimetric goals and there exists strong intrastructural competition. Inverse planning objective function should not only balance the competing objectives of different structures but also that of the individual voxels in various structures. In this work we propose to model the intrastructural tradeoff through the modulation of voxel-dependent importance factors and deal with the challenging problem of how to obtain a sensible set of importance factors with a manageable amount of computing. Instead of letting the values of voxel-dependent importance to vary freely during the search process, an adaptive algorithm, in which the importance factors were tied to the local radiation doses through a heuristically constructed relation, was developed. It is shown that the approach is quite general and the EUD-based optimization is a special case of the proposed framework. The new planning tool was applied to study a hypothetical phantom case and a prostate case. Comparison of the results with that obtained using conventional inverse planning technique with structure specific importance factors indicated that the dose distributions from the conventional inverse planning are at best suboptimal and can be significantly improved with the help of the proposed nonuniform penalty scheme. PMID- 15543793 TI - The impact of daily shifts on prostate IMRT dose distributions. AB - It is becoming common clinical practice to shift prostate patients daily based on transabdominal ultrasound (US) or imaging of implanted fiducial markers. This paper investigates the dosimetric impact of these shifts by looking at five patients shifted using an optically guided 3D US localization system and treated with IMRT. Treatment plans were generated for each patient for the following 3 cases: (1) the initial preplan, which represents the ideal case in which no shifts are necessary; (2) a postplan incorporating each day's actual shifts; and (3) a postplan in which no shifts were made but the internal organs move by the amounts indicated by daily US imaging. Results show that when daily shifts are made, doses to the target, rectal wall, and bladder wall are nearly identical to those in the preplan. Equivalent uniform dose (EUD) and tumor control probability (TCP) for these plans were also the same as for the preplans. When no shifts were made, however, the dose distributions were degraded, and the computed target EUD and TCP were lower for all five patients. The magnitude of these differences varied: for three patients, the TCP was only 1%-2% lower than for the preplan. For the other two patients, however, the EUD was reduced by more than 10%, resulting in TCP reductions of 6% and 11%. These results indicate that for a symmetric beam arrangement and properly chosenmargins, shifting the patient each day and treating without recalculating the dose is unlikely toaffect local control or the sparing of normal tissues. PMID- 15543794 TI - Determination of projection geometry from quantitative assessment of the distortion of spherical references in single-view projection radiography. AB - A method is introduced, inferring the three-dimensional (3-D) location from the 2 D radiographic shadow of an opaque spherical reference body of known radius by considering its elliptical distortion, the 2-D shadow location and a known source to-receptor distance. Three noncollinear spheres fixed to a rigid object constitute all possible degrees of freedom, i.e., the entire 3-D imaging geometry. The method may be used (a) to determine the 3-D imaging geometry from a single 2-D view and (b) to correct for foreshortening of object distances coplanar with the plane defined by the sphere triplet. Apart from the mathematical background the article describes a small feasibility experiment, performed with four different sphere diameters and a commercial dental ccd receptor system (pixel length: 0.0195 mm). The mouse-cursor based image evaluation revealed an average underestimation of the critical depth- (x-) coordinate decreasing with increasing radius (-30.3% for r=0.5 mm to 2.8% for r=2.5 mm). Intraobserver reliability (the standard deviation between three single cursor-based assessments) ranged between 0% and 8% of the actual true depth. The main source of the input error is associated with the assessment of the amount of elliptical distortion, where subpixel accuracy is demanded. Consequently, software-based automated image evaluation is required using available methods for pattern recognition and point-spread correction. Provided sufficient accuracy, the method provides an important tool for foreshortening correction, depth assessment, motion analysis, and 3-D reconstruction from two or more 2-D views. PMID- 15543795 TI - Reduction of false positives on the rectal tube in computer-aided detection for CT colonography. AB - PURPOSE: To eliminate false-positive (FP) polyp detections on the rectal tube (RT) in CT colonography (CTC) computer-aided detection (CAD). METHODS: We use a three-stage approach to detect the RT: detect the RT shaft, track the tube to the tip and label all the voxels that belong to the RT. We applied our RT detection algorithm on a CTC dataset consisting of 80 datasets (40 patients scanned in both prone and supine positions). Two different types of RTs were present, characterized by differences in shaft/bulb diameters, wall intensities, and shape of tip. RESULTS: The algorithm detected 90% of RT shafts and completely tracked 72% of them. We labeled all the voxels belonging to the completely tracked RTs (72%) and in 11 out of 80 (14%) cases the RT voxels were partially labeled. We obtained a 9.2% reduction of the FPs in the initial polyp candidates' population, and a 7.9% reduction of the FPs generated by our CAD system. None of the true positive detections were mislabeled. CONCLUSIONS: The algorithm detects the RTs with good accuracy, is robust with respect to the two different types of RT used in our study, and is effective at reducing the number of RT FPs reported by our CAD system. PMID- 15543796 TI - Feasibility study of beam orientation class-solutions for prostate IMRT. AB - IMRT is being increasingly used for treatment of prostate cancer. In practice, however, the beam orientations used for the treatments are still selected empirically, without any guideline. The purpose of this work was to investigate interpatient variation of the optimal beam configuration and to facilitate intensity modulated radiation therapy (IMRT) prostate treatment planning by proposing a set of beam orientation class-solutions for a range of numbers of incident beams. We used fifteen prostate cases to generate the beam orientation class-solutions. For each patient and a given number of incident beams, a multiobjective optimization engine was employed to provide optimal beam directions. For the fifteen cases considered, the gantry angle of any of the optimized plans were all distributed within a certain range The angular distributions of the optimal beams were analyzed and the most selected directions are identified as optimal directions. The optimal directions for all patients are averaged to obtain the class-solution. The class-solution gantry angles for prostate IMRT were found to be: three beams (0 degrees, 120 degrees, 240 degrees), five beams (35 degrees, 110 degrees, 180 degrees, 250 degrees, 325 degrees), six beams (0 degrees, 60 degrees, 120 degrees, 180 degrees, 240 degrees, 300 degrees), seven beams (25 degrees, 75 degrees, 130 degrees, 180 degrees, 230 degrees, 285 degrees, 335 degrees), eight beams (20 degrees, 70 degrees, 110 degrees, 150 degrees, 200 degrees, 250 degrees, 290 degrees, 340 degrees), and nine beams (20 degrees, 60 degrees, 100 degrees, 140 degrees, 180 degrees, 220 degrees, 260 degrees, 300 degrees, 340 degrees). The level of validity of the class-solutions was tested using an additional clinical prostate case by comparing with the individually optimized beam configurations. The difference between the plans obtained with class-solutions and patient-specific optimizations was found to be clinically insignificant. PMID- 15543797 TI - Computerized nipple identification for multiple image analysis in computer-aided diagnosis. AB - Correlation of information from multiple-view mammograms (e.g., MLO and CC views, bilateral views, or current and prior mammograms) can improve the performance of breast cancer diagnosis by radiologists or by computer. The nipple is a reliable and stable landmark on mammograms for the registration of multiple mammograms. However, accurate identification of nipple location on mammograms is challenging because of the variations in image quality and in the nipple projections, resulting in some nipples being nearly invisible on the mammograms. In this study, we developed a computerized method to automatically identify the nipple location on digitized mammograms. First, the breast boundary was obtained using a gradient-based boundary tracking algorithm, and then the gray level profiles along the inside and outside of the boundary were identified. A geometric convergence analysis was used to limit the nipple search to a region of the breast boundary. A two-stage nipple detection method was developed to identify the nipple location using the gray level information around the nipple, the geometric characteristics of nipple shapes, and the texture features of glandular tissue or ducts which converge toward the nipple. At the first stage, a rule based method was designed to identify the nipple location by detecting significant changes of intensity along the gray level profiles inside and outside the breast boundary and the changes in the boundary direction. At the second stage, a texture orientation-field analysis was developed to estimate the nipple location based on the convergence of the texture pattern of glandular tissue or ducts towards the nipple. The nipple location was finally determined from the detected nipple candidates by a rule-based confidence analysis. In this study, 377 and 367 randomly selected digitized mammograms were used for training and testing the nipple detection algorithm, respectively. Two experienced radiologists identified the nipple locations which were used as the gold standard. In the training data set, 301 nipples were positively identified and were referred to as visible nipples. Seventy six nipples could not be positively identified and were referred to as invisible nipples. The radiologists provided their estimation of the nipple locations in the latter group for comparison with the computer estimates. The computerized method could detect 89.37% (269/301) of the visible nipples and 69.74% (53/76) of the invisible nipples within 1 cm of the gold standard. In the test data set, 298 and 69 of the nipples were classified as visible and invisible, respectively. 92.28% (275/298) of the visible nipples and 53.62% (37/69) of the invisible nipples were identified within 1 cm of the gold standard. The results demonstrate that the nipple locations on digitized mammograms can be accurately detected if they are visible and can be reasonably estimated if they are invisible. Automated nipple detection will be an important step towards multiple image analysis for CAD. PMID- 15543798 TI - Large efficiency improvements in BEAMnrc using directional bremsstrahlung splitting. AB - The introduction into the BEAMnrc code of a new variance reduction technique, called directional bremsstrahlung splitting (DBS), is described. DBS uses a combination of interaction splitting for bremsstrahlung, annihilation, Compton scattering, pair production and photoabsorption, and Russian Roulette to achieve a much better efficiency of photon beam treatment head simulations compared to the splitting techniques already available in BEAMnrc (selective bremsstrahlung splitting, SBS, and uniform bremsstrahlung splitting, UBS). In a simulated 6 MV photon beam (10 x 10 cm2 field) photon fluence efficiency in the beam using DBS is over 8 times higher than with optimized SBS and over 20 times higher than with UBS, with a similar improvement in electron fluence efficiency in the beam. Total dose efficiency in a central-axis depth-dose curve improves by a factor of 6.4 over SBS at all depths in the phantom. The performance of DBS depends on the details of the accelerator being simulated. At higher energies, the relative improvement in efficiency due to DBS decreases somewhat, but is still a factor of 3.5 improvement over SBS for total dose efficiency using DBS in a simulated 18 MV photon beam. Increasing the field size of the simulated 6 MV beam to 40 x 40 cm2 (broad beam) causes the relative efficiency improvement of DBS to decrease by a factor of approximately 1.7 but is still up to 7 times more efficient than with SBS. PMID- 15543799 TI - Comparison of dose calculation algorithms in phantoms with lung equivalent heterogeneities under conditions of lateral electronic disequilibrium. AB - An extensive set of benchmark measurement of PDDs and beam profiles was performed in a heterogeneous layer phantom, including a lung equivalent heterogeneity, by means of several detectors and compared against the predicted dose values by different calculation algorithms in two treatment planning systems. PDDs were measured with TLDs, plane parallel and cylindrical ionization chambers and beam profiles with films. Additionally, Monte Carlo simulations by means of the PENELOPE code were performed. Four different field sizes (10 x 10, 5 x 5, 2 x 2, and 1 x 1 cm2) and two lung equivalent materials (CIRS, p(w)e=0.195 and St. Bartholomew Hospital, London, p(w)e=0.244-0.322) were studied. The performance of four correction-based algorithms and one based on convolution-superposition was analyzed. The correction-based algorithms were the Batho, the Modified Batho, and the Equivalent TAR implemented in the Cadplan (Varian) treatment planning system and the TMS Pencil Beam from the Helax-TMS (Nucletron) treatment planning system. The convolution-superposition algorithm was the Collapsed Cone implemented in the Helax-TMS. The only studied calculation methods that correlated successfully with the measured values with a 2% average inside all media were the Collapsed Cone and the Monte Carlo simulation. The biggest difference between the predicted and the delivered dose in the beam axis was found for the EqTAR algorithm inside the CIRS lung equivalent material in a 2 x 2 cm2 18 MV x-ray beam. In these conditions, average and maximum difference against the TLD measurements were 32% and 39%, respectively. In the water equivalent part of the phantom every algorithm correctly predicted the dose (within 2%) everywhere except very close to the interfaces where differences up to 24% were found for 2 x 2 cm2 18 MV photon beams. Consistent values were found between the reference detector (ionization chamber in water and TLD in lung) and Monte Carlo simulations, yielding minimal differences (0.4%+/-1.2%). The penumbra broadening effect in low density media was not predicted by any of the correction-based algorithms, and the only one that matched the experimental values and the Monte Carlo simulations within the estimated uncertainties was the Collapsed Cone Algorithm. PMID- 15543800 TI - Registration of central paths and colonic polyps between supine and prone scans in computed tomography colonography: pilot study. AB - Computed tomography colonography (CTC) is a minimally invasive method that allows the evaluation of the colon wall from CT sections of the abdomen/pelvis. The primary goal of CTC is to detect colonic polyps, precursors to colorectal cancer. Because imperfect cleansing and distension can cause portions of the colon wall to be collapsed, covered with water, and/or covered with retained stool, patients are scanned in both prone and supine positions. We believe that both reading efficiency and computer aided detection (CAD) of CTC images can be improved by accurate registration of data from the supine and prone positions. We developed a two-stage approach that first registers the colonic central paths using a heuristic and automated algorithm and then matches polyps or polyp candidates (CAD hits) by a statistical approach. We evaluated the registration algorithm on 24 patient cases. After path registration, the mean misalignment distance between prone and supine identical anatomic landmarks was reduced from 47.08 to 12.66 mm, a 73% improvement. The polyp registration algorithm was specifically evaluated using eight patient cases for which radiologists identified polyps separately for both supine and prone data sets, and then manually registered corresponding pairs. The algorithm correctly matched 78% of these pairs without user input. The algorithm was also applied to the 30 highest-scoring CAD hits in the prone and supine scans and showed a success rate of 50% in automatically registering corresponding polyp pairs. Finally, we computed the average number of CAD hits that need to be manually compared in order to find the correct matches among the top 30 CAD hits. With polyp registration, the average number of comparisons was 1.78 per polyp, as opposed to 4.28 comparisons without polyp registration. PMID- 15543801 TI - Removal of bone in CT angiography of the cervical arteries by piecewise matched mask bone elimination. AB - In maximum intensity projection (MIP) images of CT angiography (CTA) scans, the arteries are often obscured by bone. A bone removal method is presented that uses an additional, nonenhanced scan to create a mask of the bone by thresholding and dilation. After registration of the CTA scan and the additional scan, the bone in the CTA scan is masked. As the cervical area contains bones that can move with respect to each other, these bones are separated first using a watershed algorithm, and then registered individually. A phantom study was performed to evaluate and quantify the tradeoff between the removal of the bone and the preservation of the arteries contiguous to the bone. The influence of algorithm parameters and scan parameters was studied. The method was clinically evaluated with data sets of 35 patients. Best results were obtained with a threshold of 150 HU and a dilation of 8 in-plane voxels and two out-of-plane voxels. The mean width of the soft tissue layer, which is also masked, was approximately 1 mm. The mAs value of the nonenhanced scan could be reduced from 250 mAs to 65 mAs without a loss of quality. In 32 cases the bones were registered correctly and removed completely. In three cases the bone separation was not completely successful, and consequently the bone was not completely removed. The piecewise matched mask bone elimination method proved to be able to obtain MIP images of the cervical arteries free from overprojecting bone in a fully automatic way and with only a slight increase of radiation dose. PMID- 15543802 TI - Nitric oxide in the cerebellum of mutant mice. PMID- 15543803 TI - Purkinje cell degeneration elevates eupneic and hypercapnic ventilation in rats. AB - Previous studies have demonstrated that among cerebellar nuclei, the fastigial nucleus (FN) plays a major role in facilitation of respiration, especially during hypercapnia. Since the FN primarily receives inhibitory afferents from Purkinje cells (PCs), we hypothesized that degeneration of PCs would increase both eupneic and hypercapnic ventilation. Experiments were carried out on 20 animals (n=10 for both normal and PC-degenerated) that were divided into three groups based on the different preparations used, i.e., four pairs for the awake, three pairs for the anesthetized, and three other pairs initially for the awake and subsequently for the anesthetized. The awake normal and PCD rats were separately placed in an unrestrained whole-body plethysmograph and ventilatory parameters measured before (room air) and during hypercapnia (5% CO2 + 21% O2 + 74% N2) for 30 min. The anesthetized animals were exposed to the same level of hypercapnia applied for approximately 5 min. The results showed that both eupneic breathing and hypercapnia-induced ventilatory augmentation were significantly greater in the awake PCD rat than those observed in the normal one, primarily due to a remarkable elevation in VT with little changes in f. The same results were also observed in anesthetized preparations. A Fos protein Immunocytochemical approach was employed to determine the effect of degeneration on PCs and FN neuronal activity. Fos expression of PCs was very intensive in normal rats, but absent or diminished in PCD rats. In sharp contrast, FN Fos expression was obscure in normal rats, but very apparent in PCD rats. These data suggest that PCs play an inhibitory role in modulation of eupneic and hypercapnic ventilation via inhibiting FN neuronal activity. PMID- 15543804 TI - Neuronal nitric oxide synthase expression in cerebellar mutant mice. AB - Nitric oxide (NO) is a diffusible, multifunctional signaling molecule found in many areas of the brain. NO signaling is involved in a wide array of neurophysiological functions including synaptogenesis, modulation of neurotransmitter release, synaptic plasticity, central nervous system blood flow and cell death. NO synthase (NOS) activity regulates the production of NO and the cerebellum expresses high levels of nitric oxide synthase (NOS) in granule, stellate and basket cells. Cerebellar mutant mice provide excellent opportunities to study changes of NO/NOS concentrations and activities to gain a greater understanding of the roles of NO and NOS in cerebellar function. Here, we have reviewed the current understanding of the functional roles of NO and NOS in the cerebellum and present NO/NOS activities that have been described in various cerebellar mutant mice and NOS knockout mice. NO appears to exert neuroprotective effects at low to moderate concentrations, whereas NO becomes neurotoxic as the concentration increases. Excessive NO production can cause oxidative stress to neurons, ultimately impairing neuronal function and result in neuronal cell death. Based on their genetics and cerebellar histopathology, some of cerebellar mutant mice display similarities with human neurological conditions and may prove to be valuable models to study several human neurological disorders, such as autism and schizophrenia. PMID- 15543805 TI - Pyruvate kinase activity in cerebral hemispheres and cerebellum-brainstem of normal and hypoxic-ischemic newborn rats. AB - Energy metabolism is affected in hypoxia-ischemia. Changes in the tissue concentrations of the high-energy phosphate reserves occur early during the course of the metabolic insult and with concurrent increases in cellular ADP and AMP leading glycolysis. It has been shown that enzymes of glycolysis tend to be regulated in hypoxia and ischemia. In this study we determined pyruvate kinase (PK) activity, one of the main enzymes in glycolysis, in brain tissues of healthy (n = 15) and hypoxic-ischemic (n = 18) 7-day-old newborn rats. Left common carotid artery was ligated in the hypoxic-ischemic group and after 2 hours rats were exposed to hypoxia in a chamber at 34-36 degrees C with 8% oxygen in nitrogen. The rats were decapitated after 2 hours of hypoxia and right and left cerebral hemispheres (CH) and cerebellum-brain stem (C-BS) were removed. Pyruvate kinase activity was significantly higher in C-BSs than CHs in both groups (p < 0.00005). There was no significant difference in enzyme activities of either CHs or C-BS of hypoxic-ischemic group compared to control healthy group (p > 0.05). In conclusion, brain pyruvate kinase activity did not change in hypoxia-ischemia and suggests that PK of brain differs from other tissues where it usually increases in hypoxiaischemia. PMID- 15543806 TI - Progressive myoclonic epilepsy. AB - Progressive myoclonic epilepsies (PMEs) are a group of rare disorders characterized by the occurrence of seizures, myoclonus, and progressive neurological dysfunction. This article discusses epidemiology, genetics, pathology, clinical manifestations, EEG characteristics, methods of diagnosis and treatment of the most common causes of PME, including Unverricht-Lundborg Disease (Baltic Myoclonus), MERRF, neuronal ceroid lipofuscinosis, dentatorubropallidoluysan atrophy, Gaucher disease, Lafora disease, and sialidosis. The aim of this paper is to provide clinicians with useful clinical information in order to facilitate the diagnosis and treatment of these rare diseases. PMID- 15543807 TI - Cerebellar encoding of limb position. AB - In this paper, we review single and multijoint studies that, over the years, have provided insight on the cerebellar encoding of limb spatial position. In particular, we present support to the idea that the cerebellum integrates signals from multiple sources to encode global limb parameters. Then, we highlight the result of recent studies that analyzed quantitatively the relationships between limb end-point position and cerebellar activity. These findings suggest that the cerebellum may share with other central sensory-motor structures an anisotropic representation of limb position characterized by a strong bias along the anteroposterior axis. Finally, we speculate that this anisotropy may also subtend an internal representation of limb mechanics. PMID- 15543810 TI - Table of critical limits. PMID- 15543809 TI - The vestibulo-ocular reflex as a model system for motor learning: what is the role of the cerebellum? AB - Motor systems are under a continuous adaptive process to maintain behavior throughout developmental changes and disease, a process called motor learning. Simple behaviors with easily measurable inputs and outputs are best suited to understand the neuronal signals that contribute to the required motor learning. Considering simple behaviors, the vestibulo-ocular reflex (VOR) allows quantification of its input and motor output and its neural circuitry is among the best documented. The main candidates for plastic change are the cerebellum and its target neurons in the brainstem. This review focuses on recent data regarding the involvement of the cerebellum in VOR motor learning. Learning can be divided into that acutely acquired over a period of hours and that chronically acquired over longer periods. Both acute and chronic learning have three phases named acquisition, consolidation, and retention. The cerebellar role in retention is disputed, but there is a consensus on the need of an intact cerebellum for acquisition. Data from neuronal recording, lesion studies and transgenic mouse experiments is complex but suggests that the signal representation in the cerebellum contains aspects of both motor output and sensory input. The cerebellum apparently uses different mechanisms for acute and chronic learning as well as for increases and decreases in VOR gain. Recent studies also suggest that the signal content in the cerebellum changes following learning and that the mechanisms used for chronic adaptation involve not only changes in a head velocity component but also in the efference copy of an eye movement command signal reaching Purkinje cells. This data leads to a new conceptual framework having implications for developing theories on the role of the cerebellum in motor learning and in the search for plastic elements within the VOR circuitry. For chronic learning we hypothesize that changes in the head velocity information traveling through the circuitry occur in parallel with changes in the integrator pathway and the efference copy pathway. We further propose that these changes are necessary to maintain the broadband characteristics of the learned behavior. PMID- 15543811 TI - Table of reference intervals. PMID- 15543812 TI - Critical values for therapeutic drug levels. PMID- 15543808 TI - The effects of ethanol on the developing cerebellum and eyeblink classical conditioning. AB - In rats, developmental ethanol exposure has been used to model the central nervous system deficits associated with human fetal alcohol syndrome. Binge-like ethanol exposure of neonatal rats depletes cells in the cerebellum, including Purkinje cells, granule cells, and deep nuclear cells, and produces deficits in simple tests of motor coordination. However, the extent to which anatomical damage is related to behavioral deficits has been difficult to estimate. Eyeblink classical conditioning is known to engage a discrete brain stem-cerebellar circuit, making it an ideal test of cerebellar functional integrity after developmental ethanol exposure. Eyeblink conditioning is a simple form of motor learning in which a neutral stimulus (such as a tone) comes to elicit an eyeblink when repeatedly paired with a stimulus that evokes an eyeblink prior to training (such as mild periorbital stimulation). In eyeblink conditioning, one of the deep cerebellar nuclei, the interpositus nucleus, as well as specific Purkinje cell populations, are sites of convergence for tone conditioned stimulus and somatosensory unconditioned stimulus information, and, together with brain stem nuclei, provide the necessary and sufficient substrate for the learned response. A series of studies have shown that eyeblink conditioning is impaired in both weanling and adult rats given binge-like exposure to ethanol as neonates. In addition, interpositus nucleus neurons from ethanol-exposed rats showed impaired activation during eyeblink conditioning. These deficits are accompanied by a permanent reduction In the deep cerebellar nuclear cell population. Because particular cerebellar cell populations are utilized in well-defined ways during eyeblink conditioning, conclusions regarding the underlying neural substrates of behavioral change after developmental ethanol exposure are greatly strengthened. PMID- 15543813 TI - Cut-off and toxicity levels for drugs-of-abuse testing. PMID- 15543815 TI - Pioneering the high-tech practice: fully integrated system boosts efficiency. PMID- 15543814 TI - Why integration is key for dental office technology. AB - BACKGROUND: Information technology, or IT, devices and applications for dental practice have multiplied tremendously during the last two decades. While many of these innovations can bring significant benefits to clinical practice, integrating them into a smooth working environment is a significant challenge. METHODS: The author reviewed the level of integration of different IT-related products for dental practice and analyzed the consequences for work flow and ergonomics. He identified practical examples for different levels of integration among dental technology products. RESULTS: The integration of hardware devices and software applications in the dental care environment leaves much to be desired. Practitioners are engaged in a constant struggle to keep their "technology parks" working smoothly. Four levels of integration exist: hardware hardware, hardware-software, software-software and task-oriented information integration. While there are many examples for the successful integration of hardware devices, products tend to be less integrated in other areas. Task oriented information integration is a vision that, in general, has not been realized. CONCLUSIONS: Integration is crucial to the efficient and effective functioning of the dental office. However, practitioners cannot do much when industry does not integrate its products sufficiently. The industry, therefore, should adopt user-centered design principles, and its designers should work together to improve the integration of products from different vendors. PMID- 15543816 TI - Can technology help dentists deliver better patient care? AB - BACKGROUND: Dental practitioners are faced with a myriad of high-technology tools. Many purchase these tools without fully understanding how to use them. It is necessary to look at the simple operations of a dental office and see where these products can be of value. CONCLUSIONS AND PRACTICE IMPLICATIONS: Patient care and communication can be enhanced substantially by using several new technologies. The practitioner can use these tools to improve--not necessarily change--his or her methods. The result should be a more efficient and productive dental practice. PMID- 15543817 TI - The personal touch: how technology can strengthen the bonds between patients, professionals. PMID- 15543818 TI - Integrated office technology: how technology can help improve office efficiency. AB - BACKGROUND: While innovation in the technology industry has produced ever more advanced equipment specifically for dentistry, it is important to understand how these advances actually improve internal operations in a dental practice. OVERVIEW: The overall trend in general industry in the United States is to use technology as an instrument in enhancing productivity, and dentistry is no exception. This increased use of technology does not necessarily threaten the dentist/patient relationship. While the benefits of technology in dentistry are real, costs are not limited to just the price of the equipment itself. Each technology purchase should be evaluated on the basis of a thorough cost-benefit analysis. CONCLUSIONS AND PRACTICE IMPLICATIONS: Though dentistry is essentially a personal service, selective use of technology can benefit patient care as well as contain costs. Dentists should consider technology as a partner in their practices. PMID- 15543819 TI - Designing your office for technology. AB - BACKGROUND: To practice efficiently, dentists need to consider the successful integration of technologies, which can benefit their practice of dentistry. The physical environment of the office must be developed to accommodate not only the appropriate placement of computer hardware and high-tech dental devices, but their interconnectivity, as well. CONCLUSION: Dentists need to make appropriate decisions regarding the types of technology they choose to integrate into their offices, and they need to understand how the technology will be installed and integrated. An office designed to optimize the use of technology will produce ongoing benefits for dentists, their staff members and their patients throughout the lives of their practices. PRACTICE IMPLICATIONS: A dentist's practice must be planned to accommodate networks of systems hidden below floors, above ceilings and within walls, as well as to support and connect diverse technology items throughout the office. PMID- 15543820 TI - Putting technology in place successfully. AB - BACKGROUND: This article discusses how to integrate clinical and administrative workstations into your dental practice from the planning phase to the implementation phase. The author discusses the costs that are associated with integrating technology, as well as the hardware components and configuration. He then discusses in greater detail the core clinical technologies and how they tie in together to facilitate building a cohesive digital patient record. CONCLUSIONS: There are no shortcuts to successfully integrating technology into a dental practice. A significant commitment of time, energy and money is a prerequisite to building a secure and reliable computer network that incorporates all clinical and administrative applications. PRACTICE IMPLICATIONS: Technology is reinventing the world, and dentists need to keep pace with the people they serve. These new and not-so-new technologies will enhance dental services and productivity, which ultimately will raise the bar for the standard of care in dentistry. PMID- 15543821 TI - Viruses of mass destruction. PMID- 15543822 TI - Fixing Social Security. PMID- 15543823 TI - Both candidates are on drug trip. PMID- 15543824 TI - Will Merck survive Vioxx? PMID- 15543825 TI - Variations on a theme. Dartmouth studies suggest that more care isn't always better, with better hospitals using fewer resources, physician visits. AB - More care isn't always better, according to new studies, which arrive as pay-for performance plans are taking hold. WellPoint's Leonard Schaeffer, left, said the studies prompt questions about whether health plans should pay hospitals with high-usage patterns more if their outcomes are no better than hospitals with less use. PMID- 15543827 TI - Final farewell. Osteopathic Medical Center of Texas closes its doors. PMID- 15543826 TI - Trail of words, numbers. Fraud lawsuit targets transcription giant MedQuist. PMID- 15543828 TI - Deep sleep? Hospitals shouldn't skimp on anesthesia: JCAHO. PMID- 15543829 TI - Fall guys. Healthcare's new police dramas are about quality and reality shows. PMID- 15543830 TI - Making the vision a reality. Implementing a national IT initiative to require a highly skilled workforce. PMID- 15543831 TI - Lining up in the lobby. As work speeds up--and money is spent--on healthcare IT, vendors, trade groups want to make sure their voices are heard. PMID- 15543832 TI - Germ warfare. Vaccines, other products join a growing arsenal in fight against hospital-acquired infections. PMID- 15543833 TI - Tympanostomy tubes: an overview. PMID- 15543834 TI - Tympanostomy tubes: a review of recent studies. PMID- 15543835 TI - Ototopical agents are superior to systemic therapy for the treatment of acute and chronic otitis media. PMID- 15543836 TI - Summary of the findings of a joint consensus panel on the use of potentially ototoxic ototopicals. PMID- 15543837 TI - New developments in our understanding of ototoxicity. PMID- 15543838 TI - Absorption of intratympanic topical antibiotics. PMID- 15543839 TI - Acute otitis externa: treatment perspectives. PMID- 15543840 TI - Treatment of otologic disease in Australian Aboriginal children. PMID- 15543842 TI - Texas Health and Human Services realign. PMID- 15543843 TI - 2005-2010 Texas State Health Plan. Innovative approaches to health workforce planning in Texas. PMID- 15543844 TI - The Nobel Prize in Physiology or Medicine in 2003 to Paul C. Lauterbur, Peter Mansfield for magnetic resonance imaging. AB - Imaging of human internal organs by exact and non-invasive methods is very important for medical diagnosis, treatment and follow-up. 2003 Nobel Laureates in Physiology or Medicine have made original discoveries in the use of magnetic resonance to visualize different structures. Their discoveries have led to the development of modern magnetic resonance imaging, MRI, which represents a breakthrough in medical diagnostics and research. Atomic nuclei within a strong magnetic field rotate with a frequency depending on the strength of the magnetic field. Their energy can increase if they absorb radio waves with the same frequency (resonance). When atomic nuclei return to their previous energy status, radio waves are emitted. These discoveries were awarded the Nobel Prize in Physics in 1952. During the following decades, magnetic resonance was used mainly for studies of the chemical structure of substances. In the beginning of the 1970s, 2003 Nobel Laureates made pioneering contributions, which later led to the applications of magnetic resonance in medical imaging. Paul Lauterbur (born 1929), Urbana, Illinois, USA, discovered the possibility to create a two dimensional picture by introducing gradients to a magnetic field. Analysing the characteristics of the emitted radio waves he could determine their origin. This fact made it possible to build up a two-dimensional picture of structures that could not be visualized by other methods. Peter Mansfield (born 1933), Nottingham, England, further developed the utilization of gradients in the magnetic field. He showed, that the signals could be mathematically analysed, which made it possible to develop a useful imaging technique. Mansfield also showed how extremely fast imaging could be. It became technically possible within a decade. Magnetic resonance imaging, MRI, is now a routine method in medical diagnostics. Worldwide, more than 60 million investigations are performed each year, and the method is still rapidly evolving. MRI is often superior to other imaging techniques and significantly improves diagnostics in many diseases. MRI has replaced several invasive methods of examination and in this way has reduced the risk and discomfort of many patients. (Fig. 3, Ref. 3.). PMID- 15543845 TI - Blood platelets decrease concentration of reactive oxygen species produced by polymorphonuclear leukocytes. AB - BACKGROUND: Reactive oxygen species produced by polymorphonuclear leukocytes (PMNL) participate substantially in vascular injury induced by ischaemia and reperfusion. Blood platelets, accumulated simultaneously with PMNL may modulate this process. OBJECTIVE: To compare effects of resting and completely stimulated platelets on PMNL-derived oxidants. METHODS: Autologous human platelets and PMNL were co-incubated in the physiological cell ratio 50:1, and the formation of reactive oxygen species was detected by luminol- and isoluminol-enhanced chemiluminescence methods. To compare effects of platelets at different degrees of their activation, FMLP (selective PMNL stimulus) and Ca2+-ionophore A23187 (activates both PMNL and platelets) were used as chemiluminescence stimuli. The liberation of serotonin from platelets was estimated fluorometrically. RESULTS: Both stimulated and non-stimulated platelets inhibited PMNL chemiluminescence. However, while the decreasing effect of resting platelets disappeared at increased extracellular peroxidase concentration, the inhibition of chemiluminescence by activated platelets became even more pronounced after the addition of peroxidase and was accompanied by liberation of serotonin. The concentrations of serotonin released from platelets were sufficiently high to inhibit PMNL chemiluminescence. CONCLUSION: The obtained data indicate that by interference of platelets with peroxidase liberation from PMNL and the scavenging effect of platelet serotonin, resting and stimulated platelets might be respectively operative in inhibiting chemiluminescence. Since the presence of blood platelets in the proximity of PMNL effectively decreased the concentration of reactive oxygen species, platelets may represent a unique protective mechanism, active only in case of emergency and selectively at sites exposed to toxic effects of reactive oxygen species. (Tab. 1, Fig. 4, Ref. 42.). PMID- 15543846 TI - Cardiovascular risk factors in young Gypsy population. AB - Lipid and non-lipid cardiovascular risk parameters (cholesterol, HDL- and LDL cholesterol, triglycerides, homocysteine, C-reactive protein, insulin resistance) and data about blood pressure, smoking, body mass index were assessed in two ethnic groups aged 19-35 years--the Gypsy group (n=122) and the Slovak group (n=137) of two regions with a high density of Gypsy population. In the Gypsy group, the values of triglycerides, atherogenic index, insulin, insulin resistance were significantly increased and the level of HDL-cholesterol was significantly decreased. The risk value of atherogenic index was found in 27 % of Gypsy vs 13 % of majority subjects, and 28 % vs 24 % of subjects had hypertriglyceridemia. Risk value of insulin resistance (HOMA) was presented in 11 % of the Gypsy vs 5 % of the majority group. More obese subjects (20 % vs 8 %), more smokers (55 % vs 25 %) and more subjects with low education (85 % vs 27 %) were recorded in the minority group. The greater occurrence of dyslipidemia, obesity and insulin resistance in young Gypsy subjects is influenced with lifestyle (nutrition /prevalence of animal fat consumption, low consumption of food with low glycemic index and soluble fibre/, smoking, low physical activity) as well as low educational status. (Tab. 2, Ref. 22.). PMID- 15543847 TI - Macroenzymes and their clinical significance. AB - Macroenzymes are serum enzymes that have higher molecular mass than the corresponding enzyme normally found in serum under physiologic or pathophysiologic conditions. They are formed either by self-polymerization or by association with other serum components. Because they are cleared much more slowly than the usual enzymes, macroenzymes accumulate in the plasma and thus cause an increase in the activity of the corresponding enzyme in blood samples. Biochemically macroenzymes can be classified into two groups (macroenzymes type 1 and type 2). The importance of macroenzymes in this time is mainly in the possibility of misinterpretation of blood plasma enzyme activities. Macroenzymes are interesting also because of their association with several diseases and are being investigated as possible diagnostic markers. (Tab. 2, Ref. 20.). PMID- 15543848 TI - Central airways stenoses management--a single institution experience. AB - Tracheal stenosis is a serious, life-threatening disease with an increasing tendency. The number of complicated tracheal lesions, where resection and anastomosis can not be performed, still increases and the situation requires solution by endoprosthesis. Consequent the management of such complicated obstructive tracheal lesions is individual and time-consuming. The main objective of this study is to review the single institution experience with central airways stenosis treatment and to define the role of endotracheal stenting in tracheal reconstruction surgery. This study presents the retrospective analysis of tracheal stenosis reconstruction by means of our own modification of Montgomery T tube. (Tab. 3, Fig. 3, Ref. 12.) PMID- 15543849 TI - Radio surgery: a new tool in proctology practice. AB - BACKGROUND: An apparatus that generates ultra-high frequency current waveform [radiofrequency] has emerged as a new tool to deal with many of the common ano rectal conditions. OBJECTIVE: The purpose of this paper is to enumerate the benefits of radiofrequency with emphasis on its advantages over conventional procedures. Based on extensive personal experience of using radiofrequency surgery in the performance of various anal pathologies, an attempt is made here to relay in narrative form the procedures from a "how-we-do-it" perspective. CONCLUSION: Our experience indicates that radiofrequency surgery not only facilitates but also improves the performance of surgical procedures in treating anal fistula, hemorrhoids, pilonidal sinus, anal papillae, anal polyps, anal warts, anal condylomas, anal antibiomas and papillomas. It significantly shortens operative time, causes negligible intra-operative bleeding, facilitates faster recovery by allowing rapid healing and minimizes the chances of postoperative complications. It permitted us to perform most of the above procedures as a day care surgery. Add to this, its cost-effectiveness coupled with better patient acceptance, radiofrequency surgery, applied judicially, could significantly improve the performance of a proctologist. The technique could be effectively applied in conjunction with conventional maneuvers to better the results of these procedures. Further studies, nonetheless, are called for to define long-term benefits of the treatment. (Tab. 4, Fig. 1, Ref: 36.) PMID- 15543850 TI - Lipid peroxidation in men after dietary supplementation with a mixture of antioxidant nutrients. AB - Antioxidants and antioxidant enzymes protect living organisms against the attack of reactive oxygen species. An adequate daily intake of the individual antioxidants is therefore important to prevent the cells against oxidative damage. We investigated the effect of a modest dietary supplementation with a mixture of antioxidant nutrients (100 mg vitamin E, 100 mg vitamin C,6 mg beta carotene and 50 microg of selenium per day) for 3 months on the plasma antioxidant capacity and indices of oxidative stress. Two groups of middle-age men were selected: group 1 with survivors of myocardial infarction (MI), and group 2 with clinically normal controls. The values of total antioxidant capacity of plasma (FRAP) significantly increased after supplementation with antioxidants in the both groups. Markers of in vivo lipid peroxidation, plasma malondialdehyde (MDA) and conjugated diene (CD) levels significantly decreased in the both supplemented groups. MDA and CD values were significantly higher at baseline in the group of survivors of myocardial infarction when compared with the group of healthy men. The results demonstrate that short-term and modest supplementation with a mixture of antioxidant nutrients improves antioxidative capacity and reduces products of lipid peroxidation in plasma. Since a more pronounced effect was observed within the group of survivors of myocardial infarction, a recommendation of antioxidant supplements seems appropriate for patients with a history of cardiovascular disease. (Fig. 1, Ref. 19.) PMID- 15543851 TI - Students' research and scientific activity at the Faculty of Medicine, Comenius University in Bratislava. AB - The Faculty of Medicine of the Comenius University in Bratislava commemorated the 85th Anniversary of its foundation. In the context of this anniversary the authors draw attention to an important component of the scientific life at the Faculty of Medicine, Comenius University--namely the Students scientific and research activity "SSARA." The commencement of SSARA is related to the Youth creativity contest. The first Faculty conference was held in 1950 as a all faculty activity. One would say this conference has started the students scientific work at the faculty. In 1956 was the Students scientific board (SSB) instituted, headed by the Board of SSB. In 1967/68 a joint conference with JMF CU took place in Martin. At present one can say that SSARA at the Faculty of Medicine CU, Bratislava inspired not only faculties of CU, but also other faculties of the former Czechoslovakia to start with this activity. SSARA has become an important extension of the study and a preparation for diploma thesis. In consequence with application of credit forms of study we are in preparation of conditions allowing SSARA to become a part of optional forms of study. (Ref. 15.) PMID- 15543852 TI - A day in the life of an emergency nurse! PMID- 15543853 TI - Shared dreams, shared experiences. Interview by Jennifer Isaac. PMID- 15543855 TI - Effects of concentrated ambient particles on normal and hypersecretory airways in rats. AB - Epidemiological studies have reported that elevated levels of particulate air pollution in urban communities are associated with increases in attacks of asthma based on evidence from hospital admissions and emergency department visits. Principal pathologic features of chronic airway diseases, like asthma, are airway inflammation and mucous hypersecretion with excessive amounts of luminal mucus and increased numbers of mucus-secreting cells in regions of the respiratory tract that normally have few or no mucous cells (ie, mucous cell metaplasia). The overall goal of the present project was to understand the adverse effects of urban air fine particulate matter (PM2.5; < or = 2.5 pm in aerodynamic diameter)* on normal airways and airways compromised with airway inflammation and excess mucus. Our project was specifically designed to (1) examine the chemical and physical characteristics of PM2.5 and other airborne pollutants in the outdoor air of a local Detroit community with a high incidence of childhood asthma; (2) determine the effects of this community-based PM2.5 on the airway epithelium in normal rats and rats compromised with preexisting hypersecretory airway diseases (ie, animal models of human allergic airway disease--asthma and chronic bronchitis); and (3) identify the chemical or physical components of PM2.5 that are responsible for PM2.5 -induced airway inflammation and epithelial alterations in these animal models. Two animal models of airway disease were used to examine the effects of PM2.5 exposure on preexisting hypersecretory airways: neutrophilic airway inflammation induced by endotoxin challenge in F344 rats and eosinophilic airway inflammation induced by ovalbumin (OVA) challenge in BN rats. A mobile air monitoring and exposure laboratory equipped with inhalation exposure chambers for animal toxicology studies, air pollution monitors, and particulate collection devices was used in this investigation. The mobile laboratory was parked in a community in southwestern Detroit during the summer months when particulate air pollution is usually high (July and September 2000). We monitored the outdoor air pollution in this community daily, and exposed normal and compromised rats to concentrated PM2.5 from this local urban atmosphere. Rats in the inhalation studies were exposed for 1 day or for 4 or 5 consecutive days (10 hours/day) to either filtered air (controls) or concentrated ambient particles (CAPs) delivered by a Harvard ambient fine particle concentrator. Rats were killed 24 hours after the end of the exposure. Biochemical, morphometric, and molecular techniques were used to identify airway epithelial and inflammatory responses to CAPs. Lung lobes were also either intratracheally lavaged with saline to determine cellular composition and protein in bronchoalveolar lavage fluid (BALF) or removed for analysis by inductively coupled plasma-mass spectrometry (ICPMS) to detect retention of ambient PM2.5--derived trace elements. The Harvard concentrator effectively concentrated the fine ambient particles from this urban atmosphere (10-30 times) without significantly changing the major physicochemical features of the atmospheric particles. Daily CAPs mass concentrations during the 10-hour exposure period (0800-1800) in July ranged from 16 to 895 microg/m3 and in September ranged from 81 to 755 microg/m3. In general, chemical characteristics of ambient particles were conserved through the concentrator into the exposure chamber. Single or repeated exposures to CAPs did not cause adverse effects in the nasal or pulmonary airways of healthy F344 or BN rats. In addition, CAPs related toxicity was not observed in F344 rats pretreated with bacterial endotoxin. Variable airway responses to CAPs exposure were observed in BN rats with preexisting allergic airway disease induced by OVA sensitization and challenge. Only OVA-challenged BN rats exposed to CAPs for 5 consecutive days in September 2000 had significant increases in airway mucosubstances and pulmonary inflammation compared to saline-challenged/air-exposed control rats. OVA challenged BN rats that were repeatedly exposed to CAPs in July 2000 had only minor CAPs-related effects. In only the September 5-day exposure protocol, PM2.5 trace elements of anthropogenic origin (La, V, and S) were recovered from the lung tissues of CAPs-exposed rats. Recovery of these specific trace elements was greatest in rats with OVA-induced allergic airway disease. Additional laboratory experiments using intratracheal instillations of ambient PM2.5 samples were performed to identify bioactive agents in the CAPs to which rats had been exposed in the inhalation exposure component. Because the most pronounced effects of CAPs inhalation were found in BN rats with OVA-induced allergic airways exposed in September, we used ambient PM2.5 samples that were collected on 2 days during the September CAPs inhalation exposures to use for instillation. Ambient PM2.5 samples were collected, fractionated into soluble and insoluble species, and then compared with each other and with total PM2.5 for their effects in healthy BN rats and those with OVA-induced allergic airway disease. Intratracheal instillation of the insoluble fraction of PM2.5 caused mild neutrophilic inflammation in the lungs of healthy rats. However, total PM2.5 or the soluble or insoluble fractions instilled in rats with OVA-induced airway inflammation did not enhance the inflammation or the airway epithelial remodeling that was evident in some of the BN rats exposed to CAPs by inhalation. Therefore, the results from this instillation component did not suggest what fractions of the CAPs may have been responsible for enhancing OVA-induced airway mucosubstances and pulmonary inflammation observed in the inhalation exposure component. In summary, inhaled CAPs-related pulmonary alterations in the affected OVA-challenged rats appeared to be related to the chemical composition, rather than the mass concentration, to which the animals were exposed. Results of the trace element analysis in the lungs of CAPs-exposed BN rats exposed in September suggested that air particles derived from identified local combustion sources were preferentially retained in allergic airways. These results demonstrate that short-term exposures to CAPs from this southwestern Detroit community caused variable responses in laboratory rats and suggest that adverse biological responses to ambient PM2.5 may be associated more closely with local sources of particles and weather patterns than with particle mass. PMID- 15543856 TI - Efficiency considerations for the purely tapered interference fit (TIF) abutments used in dental implants. AB - A tapered interference fit provides a mechanically reliable retention mechanism for the implant-abutment interface in a dental implant. Understanding the mechanical properties of the tapered interface with or without a screw at the bottom has been the subject of a considerable amount of studies involving experiments and finite element (FE) analysis. In this paper, approximate closed form formulas are developed to analyze the mechanics of a tapered interference fit. In particular, the insertion force, the efficiency, defined as the ratio of the pull-out force to insertion force, and the critical insertion depth, which causes the onset of plastic deformation, are analyzed. It is shown that the insertion force is a function of the taper angle, the contact length, the inner and outer radii of the implant, the static and the kinetic coefficients of friction, and the elastic modulii of the implant/abutment materials. The efficiency of the tapered interference fit, which is defined as the ratio of the pull-out force to insertion force, is found to be greater than one, for taper angles that are less than 6 deg when the friction coefficient is 0.3. A safe range of insertion forces has been shown to exist. The lower end of this range depends on the maximum pull-out force that may occur due to occlusion in the multiple tooth restorations and the efficiency of the system; and the upper end of this range depends on the plastic deformation of the abutment and the implant due to interference fit. It has been shown that using a small taper angle and a long contact length widens the safe range of insertion forces. PMID- 15543857 TI - Quantitative examination of a perfusion microscope for the study of osmotic response of cells. AB - The perfusion microscope was developed for the study of the osmotic response of cells. In this microscope, the cells are immobilized in a transparent chamber mounted on the stage and exposed to a variety of milieus by perfusing the chamber with solutions of different concentrations. The concentration of the supplied solution is controlled using two variable-speed syringe pumps, which supply an isotonic solution and a hypertonic solution. Before using this system to characterize the osmotic response of cells, the change in the concentration of NaCl solution flowing through the chamber is examined quantitatively using a laser interferometer and an image processing technique. The NaCl concentration is increased from an isotonic condition to a hypertonic condition abruptly or gradually at a given constant rate, and decreased from a hypertonic condition to an isotonic condition. It is confirmed that the concentration is nearly uniform in the cross direction at the middle of the chamber, and the change in the NaCl concentration is reproducible. The average rate of increase or decrease in the measured concentration agrees fairly well with the given rate when the concentration is changed gradually at a constant rate. The rate of the abrupt change is also determined to be the highest limit achieved by the present method. As the first application of using the perfusion microscope for biological studies, the volume change of cells after exposure to a hypertonic solution is measured. Then, the hydraulic conductivity of the cell membrane is determinedfrom the comparison of the volume change between the experiment and the theoretical estimation for the measured change in the NaCl concentration of the perfused solution. PMID- 15543858 TI - The effect of airway wall motion on surfactant delivery. AB - Soluble surfactant and airway surface liquid transport are examined using a mathematical model of Marangoni flows which accounts for airway branching and for cyclic airway stretching. Both radial and longitudinal wall strains are considered. The model allows for variation of the amplitude and frequency of the motion, as may occur under a variety of ventilatory situations occurring during surfactant replacement therapy. The soluble surfactant dynamics of the thin fluid film are modeled by linear sorption. The delivery of surfactants into the lung is handled by setting the proximal boundary condition to a higher concentration compared to the distal boundary condition. Starting with a steady-state, nonuniform, surfactant distribution, we find that transport of surfactant into the lung is enhanced for increasing strain amplitudes. However, for fixed amplitude, increasing frequency has a smaller effect. At small strain amplitudes, increasing frequency enhances transport, but at large strain amplitudes, increasing cycling frequency has the opposite effect. PMID- 15543859 TI - Low reynolds number viscous flow in an alveolated duct. AB - Flow visualization studies and supplementary numerical simulations are carried out on slow flow through a model alveolated duct. The results reveal that the type of flow that develops in the alveoli, or cavities, is controlled by the ratio of the depth to the width of the cavity and by the ratio of cavity volume to duct volume. While weak, the slowly rotating flow in the cavity is thought to be important to the convective transport of heat and mass transfer to, or from, the walls of the cavity. The relevance of these finding to particle transport and deposition deep in the lung is discussed. PMID- 15543860 TI - Wall shear-rate estimation within the 50cc Penn State artificial heart using particle image velocimetry. AB - Particle image velocimetry (PIV) has been gaining acceptance as a routine tool to evaluate the flow fields associated with fluid mechanical devices. We have developed algorithms to investigate the wall shear-rates within the 50cc Penn State artificial heart using low magnification, conventional particle image velocimetry (PIV). Wall shear has been implicated in clot formation, a major post implant problem with artificial hearts. To address the issues of wall scattering and incomplete measurement volumes, associated with near wall measurements, we have introduced a zero masking and a fluid centroid shifting technique. Simulations using different velocity fields were conducted with the techniques to assess their viability. Subsequently, the techniques were applied to the experimental data collected. The results indicate that the size of the interrogation region should be chosen to be as small as possible to maximize resolution while large enough to ensure an adequate number of particles per region. In the current study, a 16 x 16 interrogation window performed well with good spatial resolution and particle density for the estimation of wall shear rate. The techniques developed with PIV allow wall shear-rate estimates to be obtained from a large number of sites at one time. Because a planar image of a flow field can be determined relatively rapidly, PIV may prove useful in any preliminary design procedure. PMID- 15543861 TI - Pulsatile flow in fusiform models of abdoiminal aortic aneurysms: flow fields, velocity patterns and flow-induced wall stresses. AB - As one important step in the investigation of the mechanical factors that lead to rupture of abdominal aortic aneurysms, flow fields and flow-induced wall stress distributions have been investigated in model aneurysms under pulsatile flow conditions simulating the in vivo aorta at rest. Vortex pattern emergence and evolution were evaluated, and conditions for flow stability were delineated. Systolic flow was found to be forward-directed throughout the bulge in all the models, regardless of size. Vortices appeared in the bulge initially during deceleration from systole, then expanded during the retrograde flow phase. The complexity of the vortex field depended strongly on bulge diameter In every model, the maximum shear stress occurred at peak systole at the distal bulge end, with the greatest shear stress developing in a model corresponding to a 4.3 cm AAA in vivo. Although the smallest models exhibited stable flow throughout the cycle, flow in the larger models became increasingly unstable as bulge size increased, with strong amplification of instability in the distal half of the bulge. These data suggest that larger aneurysms in vivo may be subject to more frequent and intense turbulence than smaller aneurysms. Concomitantly, increased turbulence may contribute significantly to wall stress magnitude and thereby to risk of rupture. PMID- 15543862 TI - Biodynamic response of human fingers in a power grip subjected to a random vibration. AB - BACKGROUND: Knowledge of the biodynamic response (BR) of the human hand-arm system is an important part of the foundation for the measurement and assessment of hand-transmitted vibration exposure. This study investigated the BR of human fingers in a power grip subjected to a random vibration. METHOD: Ten male subjects were used in the experiment. Each subject applied three coupling actions to a simulated tool handle at three different finger grip force levels. RESULTS AND CONCLUSIONS: The BR is practically independent of the hand coupling actions for frequencies at or above 100 Hz. Above 50 Hz, the BR is correlated to finger and hand sizes. Increasing the finger coupling force significantly increases the BR. Therefore, hand forces should be measured and used when assessing hand transmitted vibration exposure. The results also show that under a constant velocity vibration, the finger vibration power absorption at frequencies above 200 Hz is approximately twice that at frequencies below 100 Hz. This suggests that the frequency weighting specified in the current ISO 5349-1 (2001) may underestimate the high frequency effect on vibration-induced finger disorders. PMID- 15543863 TI - Design of an MRI-compatible robotic stereotactic device for minimally invasive interventions in the breast. AB - The objective of this work was to develop a robotic device to perform biopsy and therapeutic interventions in the breast with real-time magnetic resonance imaging (MRI) guidance. The device was designed to allow for (i) stabilization of the breast by compression, (ii) definition of the interventional probe trajectory by setting the height and pitch of a probe insertion apparatus, and (iii) positioning of an interventional probe by setting the depth of insertion. The apparatus is fitted with five computer-controlled degrees of freedom for delivering an interventional procedure. The entire device is constructed of MR compatible materials, i.e. nonmagnetic and non-conductive, to eliminate artifacts and distortion of the MR images. The apparatus is remotely controlled by means of ultrasonic motors and a graphical user interface, providing real-time MR-guided planning and monitoring of the operation. Joint motion measurements found probe placement in less than 50 s and sub-millimeter repeatability of the probe tip for same-direction point-to-point movements. However, backlash in the rotation joint may incur probe tip positional errors of up to 5 mm at a distance of 40 mm from the rotation axis, which may occur for women with large breasts. The imprecision caused by this backlash becomes negligible as the probe tip nears the rotation axis. Real-time MR-guidance will allow the physician to correct this error Compatibility of the device within the MR environment was successfully tested on a 4 Tesla MR human scanner PMID- 15543864 TI - Vocal fold tissue failure: preliminary data and constitutive modeling. AB - In human voice production (phonation), linear small-amplitude vocal fold oscillation occurs only under restricted conditions. Physiologically, phonation more often involves large-amplitude oscillation associated with tissue stresses and strains beyond their linear viscoelastic limits, particularly in the lamina propria extracellular matrix (ECM). This study reports some preliminary measurements of tissue deformation and failure response of the vocal fold ECM under large-strain shear The primary goal was to formulate and test a novel constitutive model for vocal fold tissue failure, based on a standard-linear cohesive-zone (SL-CZ) approach. Tissue specimens of the sheep vocal fold mucosa were subjected to torsional deformation in vitro, at constant strain rates corresponding to twist rates of 0.01, 0.1, and 1.0 rad/s. The vocal fold ECM demonstrated nonlinear stress-strain and rate-dependent failure response with a failure strain as low as 0.40 rad. A finite-element implementation of the SL-CZ model was capable of capturing the rate dependence in these preliminary data, demonstrating the model's potential for describing tissue failure. Further studies with additional tissue specimens and model improvements are needed to better understand vocal fold tissue failure. PMID- 15543865 TI - Tetrapolar measurement of electrical conductivity and thickness of articular cartilage. AB - A tetrapolar method to measure electrical conductivity of cartilage and bone, and to estimate the thickness of articular cartilage attached to bone, was developed. We determined the electrical conductivity of humeral head bovine articular cartilage and subchondral bone from a 1- to 2-year-old steer to be 1.14+/-0.11 S/m (mean+/-sd, n =11) and 0.306+/-0.034 S/m, (mean+/-sd, n =3), respectively. For a 4-year-old cow, articular cartilage and subchondral bone electrical conductivity were 0.88+/-0.08 S/m (mean+/-sd, n =9) and 0.179+/-0.046 S/m (mean+/ sd, n =3), respectively. Measurements on slices of cartilage taken from different distances from the articular surface of the steer did not reveal significant depth-dependence of electrical conductivity. We were able to estimate the thickness of articular cartilage with reasonable precision (<20% error) by injecting current from multiple electrode pairs with different inter-electrode distances. Requirements for the precision of this method to measure cartilage thickness include the presence of a distinct layer of calcified cartilage or bone with a much lower electrical conductivity than that of uncalcified articular cartilage, and the use of inter-electrode distances of the current injecting electrodes that are on the order of the cartilage thickness. These or similar methods present an attractive approach to the non-destructive determination of cartilage thickness, a parameter that is required in order to estimate functional properties of cartilage attached to bone, and evaluate the need for therapeutic interventions in arthritis. PMID- 15543866 TI - Analysis of the dynamic permeation experiment with implication to cartilaginous tissue engineering. AB - In the present study, a I-D dynamic permeation of a monovalent electrolyte solution through a negatively charged-hydrated cartilaginous tissue is analyzed using the mechano-electrochemical theory developed by Lai et al. (1991) as the constitutive model for the tissue. The spatial distributions of stress, strain, fluid pressure, ion concentrations, electrical potential, ion and fluid fluxes within and across the tissue have been calculated. The dependencies of these mechanical, electrical and physicochemical responses on the tissue fixed charge density, with specified modulus, permeability, diffusion coefficients, and frequency and magnitude of pressure differential are determined. The results demonstrate that these mechanical, electrical and physicochemical fields within the tissue are intrinsically and nonlinearly coupled, and they all vary with time and depth within the tissue. PMID- 15543867 TI - Altered mechanical behavior of epicardium under isothermal biaxial loading. AB - Most soft tissues that are treated clinically via heating experience multiaxial states of stress and strain in vivo and are subject to complex constraints during treatment. Remarkably, however, there are no prior data on changes in the multiaxial mechanical behavior of a collagenous tissue subjected to isometric constraints during heating. This paper presents the first biaxial stress-stretch data on a collagenous membrane (epicardium) before and after heating while subjected to various biaxial isometric constraints. It was found that isometric heating does not allow the increase in stiffness at low strains that occurs following isotonic heating. Moreover increasing the degree of stretch prior to heating increased the thermal stability of the tissue consistent with the concept that mechanical loading primarily affects the activation entropy, not the activation energy. PMID- 15543868 TI - Static indentation of anisotropic biomaterials using axially asymmetric indenters -a computational study. AB - Indentation has historically been used by biomechanicians to extract the small strain elastic or viscoelastic properties of biological tissues. Because of the axisymmetry of indenters used in these studies however, analysis of the results requires the assumption of material isotropy and often yields an "effective" elastic modulus. Since most biological tissues such as bone and myocardium are known to be anisotropic, the use of conventional indentation techniques for estimating material properties is therefore limited. The feasibility of using an axially asymmetric indenter to determine material directions and in-plane material properties for anisotropic tissue is explored here using finite element analysis. The load versus displacement curves as would be measured by an indenter depend on the orientation of the indenter cross section relative to the in-plane material axes, thus suggesting a method for determining the underlying material directions. Additionally, the stiffness of the tissue response to indentation is sensitive to the values of the in-plane anisotropic material properties and prestretches, and thus test results can be used to back out relevant constitutive parameters. PMID- 15543869 TI - Dependence of calculus retropulsion dynamics on fiber size and radiant exposure during Ho:YAG lithotripsy. AB - During pulsed laser lithotripsy, the calculus is subject to a strong recoil momentum which moves the calculus away from laser delivery and prolongs the operation. This study was designed to quantify the recoil momentum during Ho:YAG laser lithotripsy. The correlation among crater shape, debris trajectory, laser induced bubble and recoil momentum was investigated. Calculus phantoms made from plaster of Paris were ablated with free running Ho:YAG lasers. The dynamics of recoil action of a calculus phantom was monitored by a high-speed video camera and the laser ablation craters were examined with Optical Coherent Tomography (OCT). Higher radiant exposure resulted in larger ablation volume (mass) which increased the recoil momentum. Smaller fibers produced narrow craters with a steep contoured geometry and decreased recoil momentum compared to larger fibers. In the presence of water, recoil motion of the phantom deviated from that of phantom in air. Under certain conditions, we observed the phantom rocking towards the fiber after the laser pulse. The shape of the crater is one of the major contributing factors to the diminished recoil momentum of smaller fibers. The re entrance flow of water induced by the bubble collapse is considered to be the cause of the rocking of the phantom. PMID- 15543870 TI - Branch angle and flow into a symmetric bifurcation. AB - Arterial branches are found to be a major site for formation of arterial plaque. In this study, we investigate the role of the bifurcation angle on the flow into a symmetric bifurcation. Specially, how the changes in the bifurcation angle influences the distribution of axial wall shear in the bifurcation model. The flow in a range of branch opening half-angle of pi/25< or =theta< or =pi/4 are numerically simulated. The flow in the above models is calculated for the inlet flow Reynolds numbers of 250, 500, 1000, and 2000. It is found that at higher values of the opening angle of the bifurcation, the possibility and severity of flow separation at the appropriate wall location increases. PMID- 15543871 TI - Benchmark solution for the prediction of temperature distributions during radiofrequency ablation of cardiac tissue. AB - Several studies on radiofrequency (RF) ablation are aimed at accurately predicting tissue temperature distributions by numerical solution of the bioheat equation. This paper describes the development of a solution that can serve as a benchmark for subsequent numerical solutions. The solution was obtained using integral transforms and evaluated using a C program. Temperature profiles were generated at various times and for different convection coefficients. In addition, a numerical model was developed using the same assumptions made in obtaining the benchmark solution. Comparison of surface and axial temperature profiles shows that the two solutions match very closely, cross validating the numerical methods used in evaluating both solutions. PMID- 15543872 TI - Measurement of strain in physical models of brain injury: a method based on HARP analysis of tagged magnetic resonance images (MRI). AB - Two-dimensional (2-D) strain fields were estimated non-invasively in two simple experimental models of closed-head brain injury. In the first experimental model, shear deformation of a gel was induced by angular acceleration of its spherical container In the second model the brain of a euthanized rat pup was deformed by indentation of its skull. Tagged magnetic resonance images (MRI) were obtained by gated image acquisition during repeated motion. Harmonic phase (HARP) images corresponding to the spectral peaks of the original tagged MRI were obtained, following procedures proposed by Osman, McVeigh and Prince. Two methods of HARP strain analysis were applied, one based on the displacement of tag line intersections, and the other based on the gradient of harmonic phase. Strain analysis procedures were also validated on simulated images of deformed grids. Results show that it is possible to visualize deformation and to quantify strain efficiently in animal models of closed head injury. PMID- 15543873 TI - A method for matching the refractive index and kinematic viscosity of a blood analog for flow visualization in hydraulic cardiovascular models. AB - In this work, we propose a simple method to simultaneously match the refractive index and kinematic viscosity of a circulating blood analog in hydraulic models for optical flow measurement techniques (PIV, PMFV, LDA, and LIF). The method is based on the determination of the volumetric proportions and temperature at which two transparent miscible liquids should be mixed to reproduce the targeted fluid characteristics. The temperature dependence models are a linear relation for the refractive index and an Arrhenius relation for the dynamic viscosity of each liquid. Then the dynamic viscosity of the mixture is represented with a Grunberg Nissan model of type 1. Experimental tests for acrylic and blood viscosity were found to be in very good agreement with the targeted values (measured refractive index of 1.486 and kinematic viscosity of 3.454 milli-m2/s with targeted values of 1.47 and 3.300 milli-m2/s). PMID- 15543874 TI - Comment on "Hydrostatic pressurization and depletion of trapped lubricant pool during creep contact of a rippled indenter against a biphasic articular cartilage layer". PMID- 15543875 TI - Letter to the editor commenting on "hydrostatic pressurization and depletion of trapped lubricant pool during creep and sliding contact of a rippled indenter against a biphasic articular cartilage layer.". PMID- 15543876 TI - Key developments in orthopaedic surgery. PMID- 15543877 TI - Managing fibromyalgia. PMID- 15543878 TI - Painful bones and joints in children. PMID- 15543879 TI - Connective tissue diseases. PMID- 15543880 TI - Managing soft tissue tumours. PMID- 15543881 TI - Foot problems in children. PMID- 15543882 TI - Asthma: ten myths debunked. PMID- 15543883 TI - A GP guide to glomerulonephritis. PMID- 15543884 TI - Consulting skills: lights, cameras, action! PMID- 15543887 TI - Should nurses aim to be as autonomous as midwives? PMID- 15543888 TI - The voice of experience. PMID- 15543889 TI - PREP: making it work for you. PMID- 15543890 TI - The back to work business. PMID- 15543891 TI - Bringing health to the homeless. Interview by Janis Smy. PMID- 15543893 TI - What you need to know about ... Munchausen's syndrome. PMID- 15543892 TI - Ensuring optimum uptake of influenza immunisation. AB - Influenza poses a significant health threat to older people or those with chronic conditions. Prevention with immunisation is effective, but uptake could be hindered this year because of problems with supply. PMID- 15543894 TI - Ensuring deafened patients can participate in their health care. AB - This article discusses the general effects of adult-acquired deafness in relation to communication methods and barriers, and the difficulties posed by case conferences where professionals gather to discuss and review patients. It also examines how deafened people can be enabled to participate fully in such important meetings, how health professionals can ensure that all the information has been delivered and understood, and achieve their own objectives, and how nurses can play a key role in supporting deafened patients. PMID- 15543895 TI - Finding the best holistic tool to assess pain in the acute setting. PMID- 15543896 TI - Benchmarking Essence of Care using electronic patient records. AB - Essence of Care and its nine standards were introduced in February 2001 (Department of Health, 2001). These standards require a lot of work from benchmarking groups in clinical areas, and the translation of the national guidance into workable local practice is proving demanding for most organisations. This article describes one trust's use of an electronic toolkit for benchmarking. PMID- 15543897 TI - The recognition and treatment of acute alcohol withdrawal. AB - For most patients alcohol detoxification is a comparatively smooth process. However, on rare occasions life-threatening complications can occur. This article describes the symptoms of alcohol withdrawal that are likely to be encountered in a general hospital setting, and offers evidence-based guidance on monitoring the process and providing nursing care. PMID- 15543898 TI - Broaden your skills with a nursing secondment. PMID- 15543899 TI - Pay for performance: a business case for quality for California physician groups. PMID- 15543900 TI - Genetic variation and cardiovascular care. PMID- 15543901 TI - Metabolic syndrome: what might be occurring? PMID- 15543902 TI - Pediatric and adolescent obesity: an increasing problem. PMID- 15543903 TI - Congestive heart failure: medical management programs that work. PMID- 15543904 TI - Looking back. PMID- 15543905 TI - Oral health-related quality of life in patients treated with fixed, removable, and complete dentures 1 month and 6 to 12 months after treatment. AB - PURPOSE: This study described oral health-related quality of life (OHRQoL) before and after treatment in patients with fixed, removable, and complete dentures. MATERIALS AND METHODS: OHRQoL was measured using the German version of the Oral Health Impact Profile (OHIP-G) in a convenience sample of 107 prosthodontic patients at baseline and 1 and 6 to 12 months after treatment. The sum of OHIP-G item responses (OHIP-G49, range 0 to 196) characterized OHRQoL impairment in 42 patients treated with fixed prosthodontics, 31 patients treated with removable dentures, and 34 patients treated with complete dentures. OHIP-G49 medians were compared with the OHRQoL level in a general population sample (n = 2,026). A multivariable binomial regression analysis, controlling for the effects of baseline OHRQoL and follow-up wave, was used to compare the level of impaired OHRQoL in different prosthodontic treatment groups at follow-ups. RESULTS: OHRQoL improved in 96% of the subjects. OHIP-G49 medians reached the level of OHRQoL in the general population 1 month after treatment (fixed prosthodontics patients 6 OHIP-G units; general population subjects 5 units; removable denture patients 23 units, 15 units in general population subjects; complete denture patients 13 units, 23 units in general population subjects). OHIP-G49 medians were below population norms 6 to 12 months after treatment. In patients treated with removable/complete dentures, the expected posttreatment OHIP-G49 problem rate was 1.9 times the problem rate in patients treated with fixed prosthodontics, holding baseline OHIP-G49 and follow-up wave constant. CONCLUSION: OHRQoL changed substantially comparing pretreatment scores with 1 and 6 to 12 months of follow up in patients treated with fixed, removable, and complete dentures. PMID- 15543906 TI - The use of implant-supported prostheses in the functional and psychosocial rehabilitation of tumor patients. AB - PURPOSE: The present study investigated the residual functional constraints as well as the psychosocial rehabilitation of tumor patients following prosthetic treatment with implant-supported dentures. MATERIALS AND METHODS: A clinical examination and semistructured interview were performed in 66 of 132 consecutive patients who underwent tumor resection and subsequent implant-supported restoration between 1985 and 1997. RESULTS: Functional and psychosocial constraints were improved by between 91% (general comfort) and 47% (social reintegration) of all cases, but these constraints were never fully compensated for. Restrictions in tongue mobility, loss of sensation, and radiotherapy-induced hyposalivation led to problems in chewing and swallowing, which were hardly improved by prosthetic rehabilitation. The best subjective assessments referred to appearance, followed by masticatory improvement and denture retention. CONCLUSION: Functional impairment cannot be fully compensated by implant supported prosthodontic reconstructions, but such treatment contributes essentially to general well-being and relief of disease-related social restrictions. PMID- 15543907 TI - Effect of occlusal morphology on the accuracy of bite force measurements using thin film transducers. AB - PURPOSE: Occlusal transducer foils using piezoelectricity or pressure-dependent electric resistance are potential candidates for a measurement system for both absolute levels and changes in bite force. This preliminary in vitro study evaluated the possible usefulness of such transducer foils in the assessment of occlusal forces in centric occlusion. MATERIALS AND METHODS: Piezoelectric force transducer foils 33 microm thick were placed between pairs of teeth with the dentition in centric occlusion. Occlusal forces were recorded in five patients, who were instructed to bite as hard as possible for about 1 second. Acrylic resin casts of each pair of antagonistic teeth were aligned with the help of interocclusal records. The casts were mounted in a jig, where a defined load could be applied. The ratio of applied force and measured sensor signal permitted a set of calibration factors. RESULTS: A correction factor for each pair of teeth helped determine the ratio by which measured occlusal force exceeded actual tooth load. Differences in occlusal morphology gave rise to a wide span of correction factors (1.01 to 2.80). Steep cusp angles resulted in a wedge action that gave rise to a strong increase in occlusal forces, which were about twice as high as actual tooth load, with wide variation. CONCLUSION: This method of bite force measurement eliminated the influence of occlusal morphology by individual correction for each pair of opposing teeth. To measure the absolute load of antagonistic teeth with thin film transducer foils, one must take into account individual occlusal morphology. Forces measured with the films are a function of both actual tooth load and occlusal morphology. PMID- 15543908 TI - Clinical success of zirconium oxide posts with resin composite or glass-ceramic cores in endodontically treated teeth: a 4-year retrospective study. AB - PURPOSE: This study retrospectively evaluated zirconium oxide posts with either direct resin composite cores or indirect glass-ceramic cores after several years of clinical service. MATERIALS AND METHODS: One hundred forty-five endodontically treated teeth in 88 patients were restored with zirconium oxide posts; 87 teeth were restored with direct composite cores, and 58 were restored with Empress cores prior to crowning. Seventy-nine posts with direct composite cores in 52 patients and 34 posts with glass-ceramic cores in 19 patients were reevaluated clinically and radiographically after a mean clinical service of 57.7 months and 46.3 months, respectively. RESULTS: Periodontal probing depths remained on a healthy clinical level, and esthetics were judged excellent for the majority of teeth in both groups. In the group with direct composite cores, no failures were observed in the 79 posts that were reevaluated. In the group with indirect glass ceramic cores, three failures were observed after 42, 43, and 55 months of clinical service. All failures were due to loss of retention. In a best-case scenario posts that could not be reevaluated were considered successful, and in a worst-case scenario they were considered failures; respective success rates were 100% and 91% in the direct group and 95% and 53% in the indirect group. CONCLUSION: The observed clinical success of zirconium oxide posts with direct composite cores suggests that this method of post-and-core reconstruction is clinically promising. Zirconium oxide posts with indirect glass-ceramic cores displayed a significantly higher failure rate and a high dropout rate that precludes valid conclusions. PMID- 15543909 TI - Marginal adaptation and microleakage of Procera AllCeram crowns with four cements. AB - PURPOSE: This study investigated the effect of different cements on microleakage and marginal adaptation of porcelain crowns. MATERIALS AND METHODS: Eighty extracted molars were divided into two groups. Teeth in one group were prepared to receive Procera AllCeram crowns, whereas the other group was prepared to receive metal-ceramic crowns. Copings were made following standard techniques, and groups were divided for cementation with zinc phosphate, glass-ionomer, resin modified glassionomer, or resin cement. Specimens were subjected to thermocycling prior to microleakage testing, then sectioned. Microleakage was scored using a five-point scale; marginal adaptation was assessed with a traveling microscope. RESULTS: A significant association was found between cement type and degree of microleakage. With zinc phosphate, 76% of Procera AllCeram and 90% of metal ceramic copings exhibited extensive microleakage. With glass-ionomer, 49% of Procera AllCeram and 66% of metal-ceramic copings had 0 microleakage scores; with resin-modified glass-ionomer, 10% of Procera AllCeram and 84% of metal-ceramic copings had 0 microleakage scores. With resin cement, 34% of Procera AllCeram and 96% of metal-ceramic copings exhibited 0 microleakage. Procera AllCeram copings had a significantly larger mean marginal gap (54 microm) compared to metal ceramic (29 microm). CONCLUSION: In both types of crowns, the use of resin cement resulted in the highest percentage of 0 microleakage scores, whereas the zinc phosphate cement resulted in the highest percentage of extensive microleakage. PMID- 15543910 TI - Oral implant surfaces: Part 1--review focusing on topographic and chemical properties of different surfaces and in vivo responses to them. AB - PURPOSE: This article reviews the topographic and chemical properties of different oral implant surfaces and in vivo responses to them. MATERIALS AND METHODS: The article considers detailed mechanical, topographic, and physical characteristics of implant surfaces. Anchorage mechanisms such as biomechanical and biochemical bonding are examined. Osteoattraction and doped surfaces are discussed. RESULTS: Surface quality of an oral implant may be subdivided into mechanical, topographic, and physicochemical properties. Topographic properties are evaluated at the micrometer level of resolution. Moderately rough surfaces (Sa between 1.0 and 2.0 microm) show stronger bone responses than smoother or rougher surfaces. The majority of currently marketed implants are moderately rough. Oral implants permit bone ingrowth into minor surface irregularities biomechanical bonding or osseointegration. Additional biochemical bonding seems possible with certain surfaces. Osteoattraction is a commercial term without precise biologic correspondence. Surfaces doped with biochemical agents such as bone growth factors have been developed. CONCLUSION: Moderately roughened surfaces seem to have some clinical advantages over smoother or rougher surfaces, but the differences are small and often not statistically significant. Bioactive implants may offer some promise. PMID- 15543911 TI - Oral implant surfaces: Part 2--review focusing on clinical knowledge of different surfaces. AB - PURPOSE: This article reviews clinical knowledge of selected oral implant surfaces. MATERIALS AND METHODS: The surfaces most commonly used in clinical practice, marketed by the five largest oral implant companies, are identified; their clinical documentation was scrutinized following a strict protocol. Experimental knowledge of the surfaces is briefly summarized. Retrospective, prospective, and comparative clinical studies were analyzed separately, as were studies of implants in conjunction with bone grafts. RESULTS: TiUnite anodized surfaces are clinically documented in 1- to 2-year follow-up studies at best, with failures at about 3%. Sandblasted and acid-etched SLA surfaces are documented with good clinical results for up to 3 years. Osseotite dual acid etched implants are documented with good clinical results for up to 5 years. Frialit-2 sandblasted and etched implants are positively documented for about 3 years in one study only. The Tioblast implant is the only design documented for survival over 10 years of follow-up and success over 7 years of follow-up. CONCLUSION: Generally, oral implants are introduced clinically without adequate clinical documentation. Implant companies initiate clinical documentation after product launch. The standards of clinical reporting have improved over the years. Proper long-term reports have been published for only one surface, Tioblast. PMID- 15543912 TI - Radiographic study of changes in the mandibular condyle after implant prosthodontic rehabilitation. AB - PURPOSE: The aim of this follow-up investigation was to assess the influence of clinical variables after implant prosthodontic rehabilitation in patients without temporomandibular disorders on alterations of condylar morphology as seen radiographically. MATERIALS AND METHODS: In 167 patients, the condylar findings of flattening, erosion, osteophytes, and sclerosis were scored according to severity on rotational panoramic radiographs. Findings were followed up on images taken before and on average 5 years after implant prosthodontic rehabilitation. Changes were analyzed in relation to age, gender, initial radiologic status, and occlusal support pre- and posttreatment, according to Eichner groups, by logistic regression analysis. RESULTS: The majority of patients did not show structural alterations over the follow-up period, regardless of whether the condyle was already affected at the outset of the investigation. Condyles unaffected at the time of the first radiograph made up the majority of increases in radiographic score. Flattening and sclerosis were cumulative, whereas erosions and osteophytes had a more transient character. There was no influence of the mentioned clinical or radiographic findings on the development of changes. CONCLUSION: Over the investigated period, the incidence of all four noted findings doubled. However, the statistical methods employed could not detect any influence of patient variables or the insertion of implant-retained prosthodontics on the development of condylar changes in this specific patient group. Both assertions reflect the complexity of the topic. PMID- 15543913 TI - Influence of fatigue loading on the performance of adhesive and nonadhesive luting cements for cast post-and-core buildups in maxillary premolars. AB - PURPOSE: This study evaluated the influence of fatigue loading on the performance of an adhesive and a nonadhesive cement for cast post-and-core restorations in maxillary premolars. MATERIALS AND METHODS: The adhesive cement used was Panavia 21, a resin-based composite cement, and the nonadhesive cement was PhosphaCem/C, a zinc-oxy-phosphate cement. The coronal sections of single-rooted human maxillary premolars were removed at the level of the proximal CEJ. After endodontic treatment, a cast post and core was prepared for each tooth and cemented into the root canal with either Panavia 21 (n = 8) or PhosphaCem/C (n = 8). Half of the specimens from each cement group were exposed to fatigue loading almost perpendicular to the axial axis; the other half were used as controls. Three parallel transverse root sections were cut from each specimen and used for evaluation of the influence of fatigue loading. For each section, cement integrity was studied by SEM, and retention strength of the cemented post section was determined with a push-out test. RESULTS: For SEM evaluation and the push-out test, Panavia 21 proved significantly better than PhosphaCem/C. However, fatigue loading did not show any effect. CONCLUSION: Under the conditions of this study, fatiguing of cemented cast post-and-core restorations was not decisive as a single test to evaluate the quality of the cement. PMID- 15543914 TI - Precision of a CAD/CAM technique for the production of zirconium dioxide copings. AB - PURPOSE: The precision of a computer-aided design/manufacturing (CAD/CAM) system to manufacture zirconium dioxide copings with a predetermined internal space was investigated. MATERIALS AND METHODS: Two master models were produced in acrylic resin. One was directly scanned by the Decim Reader. The Decim Producer then manufactured 10 copings from prefabricated zirconium dioxide blocks. Five copings were prepared, aiming for an internal space to the master of 45 microm. The other five copings were prepared for an internal space of 90 microm. The second test model was used to try in the copings produced. The obtained internal space of the ceramic copings was evaluated by separate measurements of the master models and inner surfaces of the copings. The master models were measured at predetermined points with an optical instrument. The zirconium dioxide copings were measured with a contact instrument at the corresponding sites measured in the masters. RESULTS: The first group of copings had a mean internal space to the scanned master of 41 microm and of 53 microm to the try-in master. In general, the internal space along the axial walls of the masters was smaller than that along the occlusal walls. The second group had a mean internal space of 82 microm to the scanned master and of 90 microm to the try-in master. CONCLUSION: The aimed for internal space of the copings was achieved by the manufacturer. The CAD/CAM technique tested provided high precision in the manufacture of zirconium dioxide copings. PMID- 15543915 TI - Accuracy and precision of a system for assessing severity of tooth wear. AB - PURPOSE: The purpose of this study was to introduce a computer-assisted quantitative tooth wear-analyzing system and test its accuracy and precision. MATERIALS AND METHODS: An image of a horizontally projected dental arch was captured, and the facet area of each tooth was calculated using a digital image analyzer. For normalization, the facet area of the individual tooth was divided by the total area of the corresponding tooth; this result was the tooth wear score. The accuracy and precision of this method were determined through the intraclass correlation coefficient using 10 dental casts. Tooth wear on the dental casts of 50 subjects was actually measured. RESULTS: The accuracy of the tooth wear score determined by a newly trained examiner was greater than 0.85 of the intraclass correlation coefficient; this was tested against the score determined by consensus of three experienced examiners. The reproducibility within a 1-week interval was also greater than 0.85. With regard to the tooth wear score from 50 sets of dental casts, the distribution of tooth wear was generally consistent with data reported in the literature. CONCLUSION: The results suggest that the newly trained examiner can exhibit adequate accuracy and precision after appropriate training in identifying tooth wear facets. Recorded data from 50 subjects added face validity to this new system. PMID- 15543916 TI - The effect of sulcular width on the linear accuracy of impression materials in the presence of an undercut. AB - PURPOSE: This study examined the effect of gingival sulcular width, in the presence of an undercut, on the accuracy of dies poured from impressions made with different impression materials. MATERIALS AND METHODS: Three polyvinyl siloxanes (Examix, Elite, and Express), one polyether (Permadyne), and one polysulfide (Permlastic) were used to make impressions of six metal dies. The dies were machined to simulate teeth with a chamfer preparation, root convergence, and sulci of varying widths (0.10 to 0.40 mm). The impressions were poured with type IV stone, and the resulting dies were measured under a traveling microscope at the level of the finish line. The distortion of each die was calculated. RESULTS: The distortion ranged from 0.01% to 0.89%. All materials showed similar behavior: At the narrowest sulcular width (0.10 mm), the distortion was considerably larger (0.43% to 0.89%) than at the wider sulci (up to 0.20% in most cases). Examix (polyvinyl siloxane), Permlastic (polysulfide), and Permadyne (polyether) gave the most accurate dies for sulci 0.15 mm and wider. Express (polyvinyl siloxane) showed greater deformation than other materials in most sulcular widths under the conditions of this study. CONCLUSION: Accurate impressions can be expected in sulci > or = 0.15 mm in width at the level of the finish line coronal to an undercut. Impressions with 0.10-mm sulci widths are not predictable. The physical properties of the impression materials used cannot be anticipated based on their chemical group. PMID- 15543917 TI - Effect of mixing technique on shrinkage rate of one polyether and two polyvinyl siloxane impression materials. AB - PURPOSE: Different mixing techniques may affect the quality of set impression materials. Therefore, dimensional accuracy is the most important factor when constructing a passive and accurate prosthesis. In this study, observation of the shrinkage rate of one polyether and two polyvinyl siloxane impression materials, which are both commercially available for cartridge- and hand-mix techniques, was used for comparative analysis. The hypothesis was that the cartridgemix technique would produce a more precise impression. The authors also sought to confirm the hypothesis that the studied materials would remain stable after setting. MATERIALS AND METHODS: The low-viscosity polyether material was Permadyne Garant 2:1, type 3 (ESPE). The polyvinyl siloxane materials were: (1) medium-viscosity Provil Novo Medium, cartridge- and hand-mix type 2 (Heraeus Kulzer); and (2) low viscosity President, hand- and cartridge-mix type 3 (Coltene). Measurements were made according to American Dental Association specification No. 19. Ten specimens were made of each impression material; the same examiner measured each specimen 10 times. Shrinkage rates of the same materials mixed using different techniques were compared 30 minutes, 24 hours, and 72 hours after mixing. Dimensional changes at the different measuring times were also compared. The results were statistically analyzed and compared with the SPSS for Windows program package, version 11.0 (SPSS); a two-sample t test was applied to compare the mixing techniques at every measuring time; and a Friedman test was used to analyze the changes in shrinkage rate during the evaluation period (P > .05). RESULTS: Figure 1 shows the shrinkage rate data for all materials. Statistical analysis showed no significant difference at any measuring point when the mixing techniques of the polyvinyl siloxane materials were compared. However, analysis showed significant differences for both hand- and cartridge-mixed materials when measurements at 30 minutes, 24 hours, and 72 hours were compared; the shrinkage rate increased significantly as time passed. Statistical analysis of the results of the polyether material showed no significant difference at 30 minutes, but at 24 and 72 hours, the cartridge-mixed material produced a statistically higher shrinkage rate. Analysis also showed significant differences for both hand- and cartridge mixed materials when measurements at 30 minutes, 24 hours, and 72 hours were compared; shrinkage rate also increased significantly as time passed. CONCLUSION: We could not detect significant differences in dimensional changes when hand- and cartridge-mix techniques were compared at the same measuring time for the tested polyvinyl siloxane materials. The cartridge-mix technique for the polyether material showed significantly higher shrinkage at 24 and 72 hours, while the mean shrinkage rate of all six materials showed a significant time-dependent increase. PMID- 15543918 TI - Evaluation and treatment of hyperthyroidism and hypothyroidism. AB - This month's clinical practice guideline (CPG) review is on the diagnosis and treatment of hyperthyroidism and hypothyroidism as provided by the American Association of Clinical Endocrinologists. An estimated 27 million Americans have thyroid disease, and about 13 million of them are undiagnosed. The risk of thyroid disease increases with age, and women are seven times more likely than men to develop thyroid problems. Unfortunately, thyroid symptoms are usually very nonspecific and should be kept in the differential diagnosis of many clinical complaints. The format of this month's column is a little different because we received two independent manuscripts, each of which had merit. We made a decision to present both points of view and deleted repetitive material. This is an example of variable approaches one can take with reviewing and critiquing CPGs. PMID- 15543919 TI - Critique of the hyperthyroidism and hypothyroidism CPG. AB - The clinical practice guideline (CPG) for the evaluation and treatment of hyperthyroidism and hypothyroidism was developed in 2002 to update the original American Association of Clinical Endocrinologists (AACE) thyroid guidelines, which were published in 1995 (AACE Thyroid Task Force). The mission of this guideline as stated by the authors was to provide a framework for managing the diagnosis and treatment of patients with hyperthyroidism and hypothyroidism. The guidelines are intended to improve outcomes and reduce costs while stressing the importance of routine continual care provided by one physician. The guidelines are also intended to enhance physicians' awareness of subclinical hyperthyroidism and subclinical hypothyroidism, which are frequently encountered in the clinical setting and often have detrimental health consequences when untreated. The organization of the guideline is clear and easy to follow. Hyperthyroidism and hypothyroidism are described in two separate sections, each of which covers clinical features, diagnosis, laboratory evaluation, differential diagnosis, treatment, and management. (See previous article in this column.). PMID- 15543920 TI - Challenges in the assessment of erectile dysfunction and treatment with oral therapies: review with case examples. AB - PURPOSE: To review the clinical dimensions, etiology, assessment, and treatment of erectile dysfunction (ED) and to increase awareness of oral therapies for this condition. DATA SOURCES: Scientific literature, consensus guidelines, and manufacturers' product information and guidelines. CONCLUSIONS: A prevalent condition, particularly in middle-aged and elderly men, ED is vastly underreported and undertreated. Oral therapies, including phosphodiesterase type 5 inhibitors, are effective and well tolerated in many patients and represent potential first-line treatments for ED. IMPLICATIONS FOR PRACTICE: ED has been associated with both life-threatening illnesses and reduced quality of life. Improving recognition and treatment of this condition and counseling patients about ED represent important clinical opportunities. A broad and expanding array of ED treatments enable therapy to be individualized to the needs of patients and their partners. PMID- 15543921 TI - Cardiac resynchronization therapy and automatic implantable cardiac defibrillators in the treatment of heart failure: a review article. AB - PURPOSE: To review the use of cardiac resynchronization therapy (CRT) and automatic implantable cardiac defibrillators (AICDs) in heart failure (HF) patients. DATA SOURCES: Selected scientific literature. CONCLUSIONS: New developments in device therapy for HF patients are helping to decrease morbidity and mortality in this challenging patient population. CRT improves left ventricular (LV) ejection fraction, quality of life, 6-min walk distances, and New York Heart Association scores in select patients. AICDs can prevent sudden cardiac death in those who have LV dysfunction and are at risk for ventricular arrhythmias. Cardiac devices are now becoming a standard of care for those with HF who meet certain criteria. IMPLICATIONS FOR PRACTICE: Despite advances in medical therapy for treating LV dysfunction, newly diagnosed patients face a 50% mortality rate in 5 years. The natural history of HF leads to continual deterioration of function unless adverse cardiac remodeling is reversed. Until recently, the only means for improving symptoms and cardiac function has been through the optimization of standard medicines that are indicated for LV dysfunction, such as angiotensin-converting enzyme inhibitors and beta-blockers. However, not all patients benefit from medical management alone. Cardiac devices may now be considered when significant symptoms persist after standard medicines are optimized. When practitioners use a multiple-modality approach, careful patient selection based on the inclusion criteria used in the trials outlined in this article will likely lead to improved management of those with LV dysfunction. PMID- 15543922 TI - Atopic dermatitis: an overview for the nurse practitioner. AB - PURPOSE: To provide the primary care provider (PCP) with a review of the clinical presentation, aggravating factors, and basic options for treatment of atopic dermatitis (AD). DATA SOURCES: Selected scientific literature and clinical experiences of the author. CONCLUSIONS: AD is a common, chronic skin condition that is frequently seen in both children and adults. Although AD can be challenging to treat, a number of therapeutic options exist to help alleviate the pruritus and rash that accompany the disorder. IMPLICATIONS FOR PRACTICE: The PCP can manage AD in most patients by prescribing avoidance measures, good skin care, antihistamines, and conservative topical medications. Patients with more severe disease may require aggressive therapies such as phototherapy, balneo phototherapy, or systemic agents. PMID- 15543923 TI - Evidence for practice: oral contraception and risk of cervical cancer. AB - PURPOSE: The issue of continued oral contraceptive use among women with abnormal Pap smears may be controversial due to reported evidence of an increased risk of cervical cancer among long term oral contraceptive users. This article reviews the evidence concerning oral contraception and cervical cancer risk. The role of Human Papilloma virus (HPV) and proposed mechanisms for the development of cervical cancer are explained. Evidence from World Health Organization (WHO) studies and additional research evidence are discussed. Implications for clinical practice are included. DATA SOURCES: WHO commissioned studies, additional relevant studies searchable from an EBSCO database, and texts that describe the natural history of HPV. CONCLUSIONS: Epidemiological studies and biomedical research suggest a role for steroid hormones, such as oral contraceptives, in facilitating the action of HPV. However, a majority of HPV infections resolve spontaneously, despite widespread use of oral contraception. Oral contraception does not appear to increase incident HPV infection or persistence of HPV infection. The benefits of oral contraceptives appear to outweigh the risks associated with HPV facilitation. IMPLICATIONS FOR PRACTICE: NPs inform patients of abnormal Pap smears, manage the clinical care of women at risk of cervical cancer, and provide educational counseling regarding contraceptive choices. Ethical considerations include clear disclosure of potential risk. However, a risk-benefit analysis supports continued use of oral contraception among women who have abnormal Pap smears but also have access to clinical surveillance. PMID- 15543924 TI - Do nurse practitioners make a difference in provision of health counseling in hospital outpatient departments? AB - PURPOSE: This study examined whether nurse practitioners (NPs) had any impact on the type and amount of health counseling provided during patient visits to hospital outpatient departments (OPDs). DATA SOURCES: This is a secondary data analysis of the National Hospital Ambulatory Medical Care Survey from 1997 to 2000. Only patient visits to hospital OPDs were included. Rates of health counseling provided at patient visits involving an NP were compared with those without an NP. Adjusted odds ratio was reported separately for each type of health counseling provided at patient visits for nonillness care, for chronic problems, and for acute problems. CONCLUSIONS: Health counseling for diet, exercise, human immunodeficiency virus (HIV) and sexually transmitted disease (STD) prevention, tobacco use, and injury prevention are more likely to be provided at nonillness care visits involving an NP than at those not involving an NP. The presence of an NP is associated not only with higher rates of counseling for diet, exercise, and tobacco use provided at patient visits for chronic problems but also with higher rates of counseling for diet and HIV/STD prevention provided at patient visits for acute problems. IMPLICATIONS FOR PRACTICE: This study indicates an important role NPs can play in providing preventive services in outpatient hospital departments. The findings reflect the emphasis of the NP education on health counseling and patient education in clinical practice. PMID- 15543925 TI - Stating the case for overactive bladder: a nurse practicioner's perspective. AB - Overactive bladder (OAB) is a widespread syndrome characterized by symptoms of urgency, with or without urge urinary incontinence, frequency of micturition, and nocturia. Urgency is a sudden, intense, difficult-to-defer desire to pass urine and must be differentiated from normal urge or desire to void that can be deferred. It is the core symptom of OAB in the International Continence Society definition. Many misconceptions surround OAB, including that it is a normal part of aging; these contribute to the barriers to OAB diagnosis. Women are more likely than men to have OAB with incontinence; these women report diminished quality of life and are more susceptible to sleep disorders, fractures, and infections. The nurse practitioner plays an important role in diagnosing OAB during examination by eliciting specific information about symptoms from patients in their care, and in determining which treatment approaches (i.e., behavioral, pharmacologic, or a combination of both) will help them achieve optimal outcomes. PMID- 15543926 TI - Current perspectives on management of urgency using bladder and behavioral training. AB - Behavioral interventions for overactive bladder (OAB) include bladder training and behavioral training. Bladder training focuses on modifying bladder function by changing voiding habits; a bladder diary is indispensable. Behavioral training focuses on improving voluntary control rather than bladder function. It works through modification of the bladder outlet using methods, such as pelvic floor muscle training and urge suppression strategies to inhibit detrusor (bladder) muscle contraction. Behavioral methods--with biofeedback (which requires instrumentation), verbal feedback during a pelvic examination (using vaginal or anal palpation), or electrical stimulation--appear to be at least as effective in terms of reducing incontinence episodes as currently available drug therapy. However, evidence suggests that combining behavioral and drug therapy may produce better results than either approach alone. More research is needed to elucidate the role of behavioral modification in reducing urgency, frequency, and nocturia. PMID- 15543927 TI - Elevating our therapeutic expectations in overactive bladder. AB - Drug therapy for overactive bladder (OAB) most commonly includes antimuscarinic agents, which work by relaxing bladder smooth muscle through inhibition of acetylcholine receptors in the bladder. The major adverse effects with existing antimuscarinic agents are anticholinergic in nature (e.g., dry mouth, constipation, blurred vision). Oxybutynin and tolterodine have been used for several years for treatment of OAB; both are available in immediate- and extended release formulations. Fewer or less severe adverse effects are reported with the extended- versus the immediate-release formulations, with little or no difference in efficacy. Oxybutynin is also available as a transdermal patch. Trospium, which was recently approved for use in the United States, has efficacy and an incidence of dry mouth similar to existing agents but does not cross the blood-brain barrier. It requires twice-daily dosing. Two new antimuscarinic agents- darifenacin and solifenacin--are in development. Both show significantly better efficacy compared with placebo for key symptoms of OAB, including urgency. The incidence of dry mouth at the lowest effective dose is 19% for darifenacin and 8% and 14% for solifenacin (2 studies). PMID- 15543928 TI - Non-enzymatic and enzymatic lipid peroxidation of microsomes and nuclei obtained from rat liver. AB - The present study investigates in a experimental system in vitro the relationship between the non-enzymatic (ascorbate-Fe2+) and enzymatic (NADPH) lipid peroxidation in rat liver microsomes and nuclei. Chemiluminescence was measured as cpm/mg protein during 180 min at 37 degrees C. Approximately 50-55% of the fatty acids located in rat liver microsomes and nuclei are polyunsaturated with a prevalence of C18:2 n6 and C20:4 n6. The values of total light emission during the non-enzymatic and enzymatic lipid peroxidation were highest in microsomes than in nuclei. A significant decrease of C20:4 n6 and C22:6 n3 in rat liver microsomes and nuclei was observed during the lipid ascorbate-Fe2+-dependent peroxidation, whereas a significant decrease of C20:4 n6 in rat liver microsomes was observed during enzymatic lipid peroxidation. Over the time course studies, analysis of chemiluminescence in microsomes and nuclei demonstrated that the lipid peroxidation in the presence of ascorbate-Fe2+ reach a maximum at 50 and 30 min, respectively, whereas in the presence of NADPH it reachs a maximum at 20 min in both organelles. In liver microsomes and nuclei the peroxidizability index (pi) which indicates the degree of vulnerability to degradation of a selected membrane showed statistically significant differences between control versus ascorbate-Fe2+ when microsomes or nuclei were compared. Our results indicate that non-enzymatic (ascorbate-Fe2+) and enzymatic (NADPH) lipid peroxidation are operative in rat liver microsomes and nuclei but the sensitivities of both organelles to lipid peroxidation evidenced by chemiluminescence was greater in the presence of ascorbate-Fe2+ when compared with NADPH. PMID- 15543929 TI - Effect of conjugated linoleic acid on Delta-5 desaturase activity in yeast transformed with fungal Delta-5 desaturase gene. AB - Conjugated linoleic acid (CLA), a mixture of positional and geometric isomers derived from linoleic acid (LA: delta9, 12-18:2), has been shown to exhibit various biological functions based on studies using cell culture and animal models. It was postulated that the beneficial effects of CLA were exerted through suppression of production of arachidonic acid (AA; delta5,8,11,14-20:4) and consequently, production of pro-inflammatory eicosanoids. In this study, we used the baker's yeast, Saccharomyces cerevisiae, transformed with fungal delta5 desaturase gene as a model, to study whether CLA affects the activity of delta5 desaturase, a rate-limiting step which converts dihomo-gamma-linolenic acid (DGLA; delta8,11, 14-20:3) to AA. The activity of delta5-desaturase was examined in the transformed yeast incubated in a medium supplemented with DGLA and one of four different CLA isomers (c9, t11-, t10, c12-, c9, c11- and t9, t11). Results show that all four isomers were taken up readily by the yeast, and all of them suppressed the conversion of DGLA to AA. The degree of suppression, which varied significantly among four isomers was modulated by the level of CLA isomers added in the medium. Since portions of these CLA isomers could be converted to form delta5-CLA metabolites (delta5, c9, t11-, delta5, t10, c12-, delta5, c9, c11- and delta5, t9, t11-18:3), it is suggested that CLA suppressed the delta5 desaturation of DGLA to AA through substrate competition between DGLA and CLA isomers. PMID- 15543930 TI - Inhibition of Helicobacter pylori in vitro by various berry extracts, with enhanced susceptibility to clarithromycin. AB - The objective of this study was to evaluate the effects of various berry extracts, with and without clarithromycin on Helicobacter pylori. Resistance to clarithromycin by H. pylori has been reported, leading to interest in alternatives/adjuncts to therapy with clarithromycin. H. pylori American type culture collection (ATCC) strain 49503 was grown, cell suspensions were made in PBS and diluted 10-fold. One hundred microL of the suspension was then incubated for 18 h with extracts of raspberry, strawberry, cranberry, elderberry, blueberry, bilberry, and OptiBerry, a blend of the six berries, at 0.25-1% concentrations. Serially diluted cell suspensions were exposed for 1 h to clarithromycin at 15 microg/ml. Ten microl of bacterial samples from the 10(-7) dilution tube were plated and incubated for 18 h and the number of colonies were counted. Growth of H. pylori was confirmed by the CLO test. All berry extracts significantly (p < 0.05) inhibited H. pylori, compared with controls, and also increased susceptibility of H. pylori to clarithromycin, with OptiBerry demonstrating maximal effects. PMID- 15543931 TI - Intracellular localization of constitutive and inducible heat shock protein 70 in rat liver after in vivo heat stress. AB - The level and intracellular redistribution of the two nucleo-cytoplasmic members of 70 kDa heat shock protein family (constitutive, Hsc70 or Hsp73, and inducible, Hsp72) were studied in rat liver during a 24-h period after exposure of the animals to 41 degrees C whole body hyperthermic stress. The examined proteins were detected in the liver cytosol and nuclei by Western blotting and immunohistochemical staining of paraffin sections, as well as by immnocytochemical staining of isolated nuclear smears. All three techniques applied were based on the use of monoclonal antibodies recognizing both constitutive and inducible Hsp70 isoforms or only the inducible isoform, and gave consistent results. The exposure of the animals to in vivo heat stress was shown to induce the synthesis of otherwise non-existing Hsp72, rendering Hsc70 level unchanged in comparison to unstressed controls. However, immediately after the stress the intracellular redistribution of Hsc70, i.e. its nuclear accumulation, was observed. The maximal level of Hsp70 both in the cytoplasm and in the nuclei was registered 5 h after the stress, which coincided with the maximal level of Hsp72 induction. The alterations in the level and intracellular distribution of examined proteins were still noticeable 24 h after the stress. The results of this study could shed some more light on, as yet uncertain, differences between cellular functions of these two proteins, as well as on the role of the constitutive form under normal and stress conditions. PMID- 15543932 TI - Iron accumulation and iron-regulatory protein activity in human hepatoma (HepG2) cells. AB - Iron may populate distinct hepatocellular iron pools that differentially regulate expression of proteins such as ferritin and transferrin receptor (TfR) through iron-regulatory mRNA-binding proteins (IRPs), and may additionally regulate uptake and accumulation of non-transferrin-bound iron (NTBI). We examined iron regulatory protein (IRP) binding activity and ferritin/TfR expression in human hepatoma (HepG2) cells exposed to iron at different levels for different periods. Several iron-dependent RNA-binding activities were identified, but only IRP increased with beta-mercaptoethanol. With exposures between 0 and 20 microg/ml iron, decreases in IRP binding accompanied large changes in TfR and ferritin expression, while chelation of residual iron with deferoxamine (DFO) caused a large increase in IRP binding with little additional effect on TfR or ferritin expression. Cellular iron content increased beyond 4 days of exposure to iron at 20 microg/ml, when IRP binding, TfR, and ferritin had all reached stable levels. However, iron content of the cells plateaued by 7 days, or decreased with 24 h exposure to very high concentrations (>50 microg/ml) of iron. These results indicate that iron-replete HepG2 cells exhibit a narrow range of maximal responsiveness of the IRP-regulatory mechanism, whose functional response is blunted both by excessive iron exposure and by removal of iron from a chelatable pool. HepG2 cells are able to limit iron accumulation upon higher or prolonged exposure to NTBI, apparently independent of the IRP mechanism. PMID- 15543933 TI - Despite minimal hemodynamic alterations endotoxemia modulates NOS and p38-MAPK phosphorylation via metalloendopeptidases. AB - In the present study, we hypothesized that endotoxemia produces metalloendopeptidase (MEPD)-dependent generation of endothelin-1 (ET-1) and alters NOS expression correlating with p38-mitogen-activated protein kinase (MAPK) phosphorylation in thoracic aorta. Male Sprague-Dawley rats (350-400 g) were subjected to two groups randomly; sham-treated (N = 10) and lipopolysaccharide (LPS)-treated (N = 10) (E. coli LPS 2 mg/kg bolus + 2 mg/kg infusion for 30 min). The animals in each group were further subdivided into vehicle and MEPD inhibitor phosphoramidon (1 mg/kg bolus, PHOS)-treated groups. LPS produces a significant decrease in mean arterial pressure (MAP) at 2 h post endotoxemia that was blocked by PHOS. PHOS attenuated LPS-induced increase in tumor necrosis factor-alpha (TNF-alpha) concentration at 2- and 24 h post-LPS administration. LPS significantly elevated plasma concentrations of ET-1 at 2- and 24 h post endotoxemia. An upregulated preproET-1 expression following both LPS and MEPD inhibition was observed in thoracic aorta at 2 h post treatment. PHOS effectively blocked conversion of preproET-1 to ET-1 in thoracic aorta locally at 24 h post treatment in endotoxic rats. PHOS inhibited LPS-induced upregulation of inducible NOS (iNOS), downregulation of endothelial NOS (eNOS) and elevation of NO byproducts (NOx) in thoracic aorta. PHOS also blocked LPS induced upregulated p38-MAPK phosphorylation in thoracic aorta at 24 h post endotoxemia. The data revealed that LPS induces MEPD-sensitive inflammatory response syndrome (SIRS) at 2- and 24 h post endotoxemia. We concluded that inhibition of MEPD not only decreases the levels of ET-1 but also simultaneously downregulates protein expression of iNOS and phosphorylated p38-MAPK while increasing eNOS in thoracic aorta during SIRS in endotoxemia. We suggest that MEPD-dependent ET-1 and NO mechanisms may be involved in endotoxemia-induced altered p38-MAPK phosphorylation. PMID- 15543934 TI - Effects of hemodialysis on serum lipids and phospholipids of end-stage renal failure patients. AB - Patients with chronic renal failure undergoing periodic maintenance hemodialysis frequently present dyslipoproteinaemia which has been linked to the sharply increased risk of cardiovascular disease in these subjects. Reported defects on lipoprotein-related enzyme activities suggest a possible influence of hemodialysis not just to plasma lipid and lipoprotein levels but also to the composition of cell membranes. In this study, it was investigated whether the reported lipid abnormalities are accompanied by changes in serum phospholipids levels. Blood samples were obtained from 140 patients undergoing maintenance hemodialysis treatment and 122 normolipidemic healthy controls and analyzed for total serum phospholipids and their individual subclasses, as well as for total cholesterol and triglycerides, HDL-cholesterol and its subclasses. A significant decrease was observed in serum HDL cholesterol levels (p < 0.001) and its subclasses, HDL2-cholesterol (p < 0.01) and HDL3-cholesterol (p < 0.01) in patients when compared with healthy controls. A critical increase in the serum triglyceride content (p < 0.001) of patients was also observed. In addition, the serum levels of sphingomyelin (p < 0.01) and diphosphatidylglycerol (p < 0.001) were increased in the patient group, while the levels of phosphatidylcholine (p < 0.01) and phosphatidylinositol (p < 0.01) were significantly decreased in the patient group compared to healthy controls. In conclusion, this work clearly demonstrates that hemodialysis treatment contributes significantly to the dyslipidemic profile of end-stage renal failure patients by altering serum lipoprotein and phospholipids concentrations. PMID- 15543935 TI - Role and differential expression of calpastatin mRNA and protein in cultured cardiomyocytes exposed to hypoxic stress. AB - We previously proposed that the calpain-mediated proteolytic pathway is activated in cultured cardiomyocytes following exposure to hypoxia (Mol Cell Biochem 214: 47, 2000). The potential role of calpastatin, the endogenous specific inhibitor of calpain, and its expression in the hypoxic state were investigated here. Hypoxia induced the expression of two calpastatin and multiple VEGF splice variants. Although cardiomyocytes and fibroblasts responded to hypoxia differentially, both cell types exhibited hypoxia-induced calpastatin transcription. The two functional calpastatin splice variants encoding the 593- and 654-amino acid calpastatin isoforms differed only in their N-terminal leader domain sequences. In spite of the increased mRNA expression, levels of the calpastatin protein doublet were not increased, but rather slightly decreased under the hypoxic condition. Cardiac hypoxia was accompanied by preferential proteolytic cleavage of troponin I (TnI), one of the major myofibrillar proteins. Forced expression of calpastatin through an adenoviral vector effectively prevented the hypoxia- and calpain-mediated TnI proteolysis. Our results highlight the discordant expression pattern of cardiac calpastatin mRNA and protein in the hypoxic state. We suggest that although induction of calpastatin gene transcription may constitute a compensatory mechanism coping with the hypoxic stress, a sustained high calpastatin protein level appears to be essential in the intervention of the activated calpain proteolytic cascade. PMID- 15543936 TI - Increased expression of H11 kinase stimulates glycogen synthesis in the heart. AB - OBJECTIVE: H11 kinase is a serine/threonine kinase preferentially expressed in the heart, which participates in cardiac cell growth and also in cytoprotection during ischemia. A cardiac-specific transgenic mouse overexpressing H11 kinase (2 to 7-fold protein increase) has been generated, and is characterized by cardiac hypertrophy with preserved function and protection against irreversible damage during ischemia/reperfusion. In this study, we tested whether H11 kinase also participates in the metabolic adaptation to cardiac hypertrophy and ischemia. METHODS AND RESULTS: A yeast two-hybrid screen using H11 kinase as a bait in a human heart library revealed a potential interaction with phosphoglucomutase (PGM), the enzyme converting glucose 6-phosphate into glucose 1-phosphate. Interaction between H11 kinase and PGM was confirmed by co-immunoprecipitation. To test the biochemical relevance of this interaction, PGM activity was measured in the heart from wild type and transgenic mice, showing a 20% increase of Vmax in the transgenic group, without change in KM. Glycogen content was increased proportionately to the expression of the transgene, reaching a 40% increase in high-expression transgenic mice (7-fold increase in H11 kinase protein) versus wild type (p < 0.01). Increased incorporation of glucose into glycogen was coupled to a 3-fold increase in the protein expression of the glucose transporter GLUT1 in plasma membrane of transgenic mice (p < 0.01). CONCLUSION: H11 kinase promotes the synthesis of glycogen, an essential fuel for the stressed heart in both conditions of overload and ischemia. Therefore, H11 kinase represents an integrative sensor in the cardiac adaptation to stress by coordinating cell growth, survival and metabolism. PMID- 15543938 TI - A correlation between membrane cholesterol level, cell adhesion and tumourigenicity of polyoma virus transformed cells. AB - We have investigated the implications of the rise in membrane cholesterol levels on several in vitro and in vivo properties of polyoma virus transformed rat fibroblasts (PyF), with a special emphasis on alpha5beta1 integrin functions. We show that increased membrane cholesterol causes the PyF cells to change their shape and become more bipolar in appearance. These cells also show significantly higher adhesion to the cell-binding domain of fibronectin, increased localization of alpha5beta1 integrin and talin molecules in focal adhesions and a more robust actin cytoskeleton organization. PyF cells with increased membrane cholesterol show reduced growth in vitro and tumours caused by these cells in nude mice are slow growing. These changes in the growth properties of PyF cells are reversible when the cholesterol levels of PyF cells become normal. Our results suggest that changes in membrane cholesterol levels influence the growth and morphological properties of transformed cells, which can be exploited in controlling the growth of tumours in vivo. PMID- 15543937 TI - Effect of dexamethasone on moesin gene expression in rabbit bone marrow stromal cells. AB - The influence of dexamethasone on rabbit bone marrow stromal cells differentiation was studied by screening the action of dexamethasone on gene expression. Using differential display, we observed some differential amplifications. The use of five of thirteen different primers combination allowed to identify one or more differential bands. One of them was identified as moesin gene. Real-time PCR confirmed a significant reduction of moesin gene expression following dexamethasone treatment. The decrease of expression for this protein, involved in cytoskeletal organization, could explain the effects of dexamethasone treatment on bone marrow stromal cells differentiation. PMID- 15543939 TI - Bioenergetic remodeling of heart mitochondria by thyroid hormone. AB - Changes in thyroid status are associated with profound alterations in biochemical and physiological functioning of cardiac muscle impacting metabolic rate, contractility and structural hypertrophy. Using an in vivo model of chronic treatment with thyroid hormone (T4, 0.3 mg/kg/day), we evaluated how mitochondria are regulated in response to T4, and assessed the relationship of T4-induced mitochondrial biogenesis and bioenergetics to overall cardiac hypertrophy. The role of thyroid hormone in cardiac bioenergetic remodeling was addressed in rats treated with T4 for 5, 10 and 15 days. Over that time, myocardial oxygen consumption substantially increased as did cardiac hypertrophy. Myocardial levels of mitochondrial enzyme activities, mitochondrial DNA (mtDNA), specific proteins and transcript were assessed. Activity levels of respiratory complexes I-V and citrate synthase significantly increased with 15 but not with 5 or 10-day T4 treatment. Myocardial levels of mtDNA, mitochondrial proteins (e.g. cytochrome c, cytochrome b, ATPase subunits, MnSOD) and the global transcription factor PPARalpha were significantly elevated with 15-day T4. Transcript analysis revealed increased expression of transcription factors and cofactors involved in mitochondrial biogenesis including PPARalpha, mtTFA, ErbAalpha and PGC-1alpha. Our findings indicate parallel increases in myocardial mitochondrial bioenergetic capacity, oxygen consumption and markers of mitochondrial biogenesis with 15-day T4; these changes were not present with 10-day T4 even with significant cardiac hypertrophy. The marked, parallel increases in PPARalpha levels suggest its potential involvement in mediating myocardial-specific remodeling of mitochondria in response to T4. PMID- 15543940 TI - Luteolin and luteolin-7-O-glucoside from dandelion flower suppress iNOS and COX-2 in RAW264.7 cells. AB - Both reactive oxygen- and nitrogen-derived reactive species play important roles in physiological and pathophysiological conditions. Flavones, luteolin and luteolin-7-O-glucoside along with a rich plant source of both flavones, namely dandelion (Taraxacum officinale) flower extract were studied for antioxidant activity in different in vitro model systems. In this current study, luteolin and luteolin-7-O-glucoside at concentrations lower than 20 microM, significantly (p < 0.05) suppressed the productions of nitric oxide and prostaglandin E2 (PGE2) in bacterial lipopolysaccharide activated-mouse macrophage RAW264.7 cells without introducing cytotoxicity. The inhibitory effects were further attributed to the suppression of both inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression, and not reduced enzymatic activity. Similar suppression for both inducible enzymes was also found with the presence of dandelion flower extract, specifically, the ethyl acetate fraction of dandelion flower extract which contained 10% luteolin and luteolin-7-O-glucoside. PMID- 15543942 TI - Diethyldithiocarbamate can induce two different type of death: apoptosis and necrosis mediating the differential MAP kinase activation and redox regulation in HL60 cells. AB - Although apoptosis and necrosis have been considered different pathways to cell death, only one compound induces both types of cell death. Diethyldithiocarbamate (DDC) has been shown to have antioxidant or prooxidant effects in several different systems. We observed in our present study that DDC induced not only apoptosis but also necrosis depending on its dosage in HL60 premyelocytic leukemia cells. Moreover, in hypoxia cell culture conditions, DDC-induced necrotic cells decreased but DDC-induced apoptosis continued. We investigated the DDC-induced different cell death mechanisms as they are correlated with reactive oxygen species (ROS). High-dose DDC-induced necrotic cell death is thought to depend on the increase of intracellular ROS, while low-dose DDC-induced apoptosis is thought to depend on changes of the intracellular redox state by the transporting of external metal ions. There was no sequential or quantitative change of Bcl-2 family proteins in DDC-induced apoptotic or necrotic pathways. However, the mitochondrial transmembrane potential was remarkably decreased in the DDC-induced necrosis. Finally, duration of c-Jun N-terminal kinase (JNK) activation resulted in different types of cell death. PMID- 15543941 TI - Cell kinetic status of mouse lens epithelial cells lacking alphaA- and alphaB crystallin. AB - alphaA- and alphaB-crystallins are small heat shock proteins and molecular chaperones that are known to prevent non-specific aggregation of denaturing proteins. Recent work indicates that alphaA-/- lens epithelial cells grow at a slower rate than wild-type cells, and cultured alphaB-/- cells demonstrate increased hyperproliferation and genomic instability, suggesting that these proteins may exert a direct effect on the cell cycle kinetics, and influence cell proliferation. However, the cell cycle parameters of alphaA/alphaBKO (double knockout) cells have not been analyzed. Here we investigate the cell cycle kinetics of synchronized mouse lens epithelial cultures derived from wild-type and alphaA/alphaB double knockout (alphaA/alphaBKO) mice using BrdU labeling of proliferating cells, and flow cytometric analysis. We also provide data on the changing pattern of expression of HSP25, a small heat shock protein in alphaA/alphaBKO and wild-type cells during the cell cycle. Using serum starvation to synchronize cells in the quiescent G0 phase, and restimulation with serum followed by BrdU labeling and flow cytometry, the data indicated that as compared to wild-type cells, a <50% smaller fraction of the alphaA/alphaBKO cells entered the DNA synthetic S phase of the cell cycle. Furthermore, there was a delay in cell cycle transit through S phase in alphaA/alphaBKO cells, suggesting that although capable of entering S phase, the alphaA/alphaBKO cells are blocked in G1 phase, and are delayed in their cell cycle progression. Immunoblot analysis with antibodies to the small heat shock protein HSP25 indicated that although HSP25 increased in G1 phase of wild-type cells, and remained elevated on further progression through the cell cycle, HSP25 accumulation was delayed to S phase in alphaA/alphaBKO cells. These data can be interpreted to indicate that mouse lens epithelial cell progression through the cell cycle is significantly affected by expression of alphaA and alphaB-crystallin. PMID- 15543943 TI - Effect of homologous series of n-alkyl sulfates and n-alkyl trimethylammonium bromides on low molecular mass protein tyrosine phosphatase activity. AB - The effect of anionic and cationic surfactants on acid phosphatase denaturation has been extensively studied. Low molecular mass (LMr) protein tyrosine phosphatase (PTP), a key regulatory enzyme involved in many different processes in the cell, was distinctly affected by anionic (homologous series of n-alkyl sulfates (C8-C14)) and cationic (n-alkyl trimethylammonium bromides (C12-C16)) surfactants. At concentrations 10-fold lower critical micellar concentration (cmc) values, the enzyme was completely inactivated in the presence of anionic surfactants, in a process independent of the pH, and dependent on the chain length of the surfactants. Under the same conditions, the effect of cationic surfactants on the enzyme activity was pH-dependent and only at pH 7.0 full inactivation was observed at concentrations 10-fold higher cmc values. In contrast to cationic surfactants the effect of anionic surfactants on the enzyme activity was irreversible and was not affected by the presence of NaCl. Inorganic phosphate, a known competitive inhibitor of PTP, protected the enzyme against inactivation by the surfactants. Our results suggest that the inactivation of the LMr PTP by anionic and cationic surfactants involved both electrostatic and hydrophobic interactions, and that the interactions enzyme-surfactants probably occurred at or near the active site. PMID- 15543944 TI - Ventricular but not atrial electro-mechanical delay of the embryonic heart is altered by anoxia-reoxygenation and improved by nitric oxide. AB - BACKGROUND/AIM: Excitation-contraction coupling is modulated by nitric oxide (NO) which otherwise has either beneficial or detrimental effects on myocardial function during hypoxia-reoxygenation. This work aimed at characterizing the variations of electromechanical delay (EMD) induced by anoxia-reoxygenation within the developing heart and determining whether atrial and ventricular EMD are modulated by NO to the same extent. METHODS: Hearts of 4 or 4.5-day-old chick embryos were excised and submitted in vitro to normoxia (45 min), anoxia (30 min) and reoxygenation (60 min). Electrocardiogram and atrial and ventricular contractions were simultaneously recorded throughout experiment. Anoxia reoxygenation-induced chrono-, dromo-and inotropic disturbances and changes in EMD in atrium (EMDa) and ventricle (EMDv) were investigated in control hearts and in hearts exposed to 0.1, 1, 10, 50 and 100 microM of DETA-NONOate (a NO donating agent) or to 50 microM of L-NAME (a NOS inhibitor). RESULTS: Under normoxia, heart rate, PR interval, ventricular shortening velocity, EMDa and EMDv were similar in control, L-NAME-treated and DETA-NONOate-treated hearts. Under anoxia, cardiac activity became markedly erratic within less than 10 min in all groups. At the onset of reoxygenation, EMDv was increased by about 300% with respect to the preanoxic value while EMDa did not vary significatively. Compared to control conditions, L-NAME or DETA-NONOate had no influence on the negative chrono-, dromo- and inotropic effects induced by anoxia-reoxygenation. However, L-NAME prolonged EMDv during anoxia and delayed EMDv recovery during reoxygenation while 100 microM DETA-NONOate had the opposite effects. EMDa was neither affected by NOS inhibitor nor NO donor. At the end of reoxygenation, all the investigated parameters returned to their basal values. CONCLUSION: This work provides evidence that a NO-dependent pathway is involved in regulation of the ventricular excitation-contraction coupling in the anoxic-reoxygenated developing heart. PMID- 15543945 TI - Flowcytometric assessment of the effect of drugs on Giardia lamblia trophozoites in vitro. AB - The effect of albendazole, vincristine and metronidazole on the adherence, morphology, cell cycle and viability of Giardia lamblia trophozoites was evaluated. Albendazole rendered the trophozoites unable to attach which is a prerequisite for establishment of Giardia infection. The adherence assay showed that albendazole caused the detachment of trophozoites at a significantly lower concentration (0.1 microg/ml) as compared to metronidazole and vincristine (40 and 60 microg/ml, respectively) in the time period of 9 h. However, at the higher concentration (10 microg/ml) albendazole caused the similar effect within the time period of 3 h. The microscopic studies revealed that albendazole brought about gross morphological changes in G. lamblia, i.e., rounding of trophozoites and disruption of ventral adhesive disc along with vacuolation in cytoplasm. Metronidazole caused the ballooning of cytoplasm and induced the most lethal effect on trophozoites (8.17% viability) as compared to vincristine and albendazole (72.13 and 65.49% viability) as shown by flowcytometric analysis. Further, the flowcytometric findings indicated that the three drugs led to the arrest of growth of trophozoites at different stages of cell cycle. Albendazole and vincristine interfered with the formation of spindle/basal body microtubules thereby arresting the cell cycle in G2 + M phase whereas metronidazole arrested the growth in S-phase by inhibiting the DNA segregation and cell division. Hence, flowcytometry has been usefully employed to study the effect of drugs on viability and DNA of G. lamblia trophozoites in the present study. PMID- 15543946 TI - Jak2 tyrosine kinase residues glutamic acid 1024 and arginine 1113 form a hydrogen bond interaction that is essential for Jak-STAT signal transduction. AB - Angiotensin II is a well-known vasoactive peptide, but it can also act as a potent growth factor, partially through activation of the tyrosine kinase Jak2. Activated Jak2 tyrosine phosphorylates and activates members of the Signal Transducers and Activators of Transcription (STAT) family of cytoplasmic transcription factors. Recently, we demonstrated that tryptophan 1020 and glutamic acid 1024 within the Jak2 activation loop are required for Jak2 tyrosine kinase activity. Here, we sought to elucidate the requirement of glutamic acid 1024 for Jak2 function. Using molecular modeling algorithms of the Jak2 kinase domain, we identified a putative interaction between glutamic acid 1024 and an arginine at position 1113. We generated a series of charge-based substitution mutations at position 1113 and found that conversion of arginine 1113 to glutamic acid, alanine, or lysine prevented Jak2 autophosphorylation. Furthermore, mutation of arginine 1113 prevented the following angiotensin II-dependent processes from occurring: (1) Jak2 tyrosine phosphorylation, (2) Jak2/AT1receptor co-association, (3) STAT1 recruitment to the Jak2/AT1receptor complex, (4) STAT1 tyrosine phosphorylation, and (5) STAT-mediated gene expression. We determined that the interaction between glutamic acid 1024 and arginine 1113 consists of two distinct hydrogen bonds. We conclude that these hydrogen bond interactions are critical for Jak2 kinase function and subsequent angiotensin II-dependent activation of the Jak/STAT signaling pathway. PMID- 15543947 TI - The p50-p50 NF-kappaB complex as a stimulus-specific repressor of gene activation. AB - The transcription factor NF-kappaB can be activated in different forms, including transcriptional activating and repressing forms. Intestinal epithelial cells have been found to modulate the relative levels of the p65-p50 and p50-p50 NF-kappaB complexes in a number of instances, and here we show that this ratio was altered in response to dietary fiber (wheat bran) and carcinogen exposure (azoxymethane). The influence of these complexes on gene regulation was examined in more detail in cell culture models. The colon-derived HT-29 cell line likewise activated both p65-p50 and p50-p50 NF-kappaB complexes: TNF-alpha triggered a strong, sustained p65-p50 activation with lower relative levels of p50-p50, whereas IL-1beta transiently activated p65-p50 with higher relative levels of p50-p50. Transfection experiments with an NF-kappaB reporter plasmid indicated that p50 was a repressor in HT-29 cells. Increased expression of the p50-p50 dimer by an adenovirus showed that the p50-p50 dimer suppressed IL-1beta activation of endogenous genes more than 5-fold (TNF-alpha, Cox-2 and IL-8), whereas gene activation by TNF-alpha was not significantly affected. DNA binding analyses showed a number of strong p50-p50 binding sites on these promoters. The selective p50-p50 suppression of IL-1beta gene activation corresponded to the transient nature of p65-p50 activation induced by IL-1beta (in both HT-29 and Caco-2 cells). Our findings demonstrate a novel gene regulatory mechanism for the NF kappaB p50-p50 complex: a signal-specific transcriptional repression that appears to selectively inhibit stimuli that transiently activate p65-p50 complexes. PMID- 15543948 TI - Modulation of iron on mitochondrial aconitase expression in human prostatic carcinoma cells. AB - The mitochondrial aconitase (mACON) containing a [4Fe-4S] cluster is regarded as the key enzyme for citrate oxidation in the epithelial cells of human prostate. In vitro studies using the human prostatic carcinoma cells, PC-3 cells, found that both hemin and ferric ammonium citrate (FAC) significantly increased mACON enzymatic activity and gene expression. The effect of FAC on mACON was enhanced 2 fold by co-treating with ascorbic acid but blocked by co-treating with iron chelator, deferoxamine mesylate. Hemin treatments blocked 30% of citrate secretion from PC-3 cells but upregualted 2-fold of intracellular ATP biosynthesis. Results from reporter assay by using a cytomegalovirus enhance/promoter driven luciferase mRNA ligated to the iron response element (IRE) of mACON as a reporter construct demonstrated that modulation of FAC on gene translation of mACON gene is dependent on the IRE. Transient gene expression assays indicated that upregulation of mACON gene transcription by FAC may through the putative antioxidant response element (ARE) signal pathway. This study provides the first evidence of the biologic mechanism of human mACON gene translation/transcription and suggests a regulatory link between the energy utilization and the iron metabolism in human prostatic carcinoma cells. PMID- 15543949 TI - Thyroid hormone stimulates 5'-ecto-nucleotidase of neonatal rat ventricular myocytes. AB - Degradation of adenine nucleotides in myocardial cells has important physiological implications associated with the regulation of the high-energy phosphate precursor pool and the production of adenosine. Adenosine may be released as from cells or, following adenine nucleotides release, they may be metabolized and rapidly converted to adenosine via the action of an ectoenzyme cascade formed by an ATP diphosphohydrolase and a 5'-nucleotidase. Thyroid hormones are known to have profound effects on the cardiovascular system, as demonstrated by the changes accompanying both hypothyroidism and hyperthyroidism. We previously reported that thyroid hormone significantly increases the ecto-5' nucleotidase (CD73) activity and expression in C6 glioma cells culture. The object of the present study was to evaluate the extracellular adenosine production from AMP in cardiomyocytes and also the effect of (T3) on activity and expression of the enzyme, CD73. Primary cultures of rat ventricular neonatal cardiac myocytes were submitted to increasing doses of T3 for 24 h. Cell viability and purity were estimated by measuring the release of lactate dehydrogenase (LDH) activity and immunofluorescence cell staining, respectively. CD73 activity was measurement using a malachite green method and RT-PCR was used to analyze enzyme expression. T3 stimulated CD73 activity and expression of the cells, suggesting that this effect could promote an increase in adenosine formation and, therefore, has an important modulatory role in the elicitation of responses that serve to restore the tissue oxygen supply-to-demand ratio back to normal. PMID- 15543951 TI - [Increased mortality in corticosteroid treated head injuries]. PMID- 15543952 TI - [The case method teaching can be a bridge. Integrated preclinical and clinical training--examination required for study motivation]. PMID- 15543950 TI - Fasting-induced inhibition of collagen biosynthesis in rat skin. A possible role for phosphoenolpyruvate in this process. AB - Fasting is accompanied by a decrease in collagen biosynthesis. The mechanism of this phenomenon involves inhibition of prolidase activity, an enzyme that plays a key role in upregulation of collagen metabolism. The mechanism of fasting-induced inhibition of prolidase activity is not known. Phosphoenolpyruvate (PEP) is known as a strong inhibitor of prolidase activity. It exerts this effect by inhibition of the enzyme phosphorylation. Unphosphorylated prolidase is inactive. One may expect that fasting-associated increase in posphoenolpyruvate content in animal tissues may be a factor which inactivates prolidase and makes it inactive in collagen biosynthesis. We measured the levels of phosphoenolpyruvate, pyruvate, and pyruvate kinase in the skin of control and fasted rats and correlated these parameters with prolidase expression, prolidase activity and collagen biosynthesis in this tissue. Significant increase of PEP concentration (about 30%) was found in the skin of fasted rats. In the same time prolidase activity and collagen biosynthesis decreased by about 50% and 30%, respectively, compared to controls. It is known that phosphoenolpyruvate is converted to pyruvate by the action of pyruvate kinase. Since fasting significantly decreases the activity of this enzyme, one may suggest that the accumulation of PEP is caused by a reduced utilisation of this metabolite. As demonstrated by Western immunoblot analysis the decrease in prolidase activity was not accompanied by a decrease in the amount of the enzyme protein. Instead, a decrease in the enzyme phosphorylation was observed. The reduction in phosphorylation seems to be responsible for the decrease in prolidase activity. These data suggest that fasting-evoked accumulation of PEP reduces the activity of prolidase, providing a mechanism for inhibition of collagen biosynthesis in the skin. PMID- 15543953 TI - [The case method--engaging teaching method. Realistic exercises make practical training of the medical profession possible]. PMID- 15543954 TI - [Great regional differences in quality of obstetrical care]. AB - In Sweden antenatal care is free of charge, easily available and utilized by practically all pregnant women. The obstetrical care is of high quality and perinatal mortality among the lowest in the world. However, there is a potential for improvement, as evidenced by the regional differences in quality of care shown in this study. Five quality indicators of antenatal and delivery care 1998 2002 are presented for all the counties in Sweden: still birth rate, smoking habits in early pregnancy, Apgar score at five minutes, neonatal mortality, and severe perineal tear. The results were adjusted for several maternal background factors as well as for pregnancy duration. In some counties the frequency of still birth, neonatal mortality, and low Apgar score at five minutes, were two to three times higher compared to the counties with the best outcome. Differences regarding severe perineal tear and smoking during early pregnancy were also evident. PMID- 15543955 TI - [Perianal/vulvovaginal/penile streptococcal infection. A childhood disease easily diagnosed with rapid testing]. PMID- 15543956 TI - [Continuous education activities arranged by specialty associations surveyed]. PMID- 15543957 TI - [Euthanasia discussion in Lakartidningen during 100 years. Few studies but many arguments for one's conviction]. PMID- 15543958 TI - [Do physicians have time for their patients?]. PMID- 15543959 TI - [Medical associations in Stockholm: Stop compulsory referrals--they take time from patient management]. PMID- 15543960 TI - [Clinical trials: GCP is no quality guarantee]. PMID- 15543961 TI - [Before the next ticks season: vaccinate children older than 7 years living in endemic regions against TBE but let the small children get away]. PMID- 15543962 TI - [Report from the inclined plane!]. PMID- 15543963 TI - [Is severity of ADHD due to the environment?]. PMID- 15543964 TI - [A questionnaire about compulsory referrals in Stockholm]. PMID- 15543965 TI - Honoring Barry L. Karger. PMID- 15543966 TI - Barry Karger 65. PMID- 15543967 TI - Strategies for shotgun identification of post-translational modifications by mass spectrometry. AB - The global identification of post-translationally modified proteins is a difficult challenge that is currently being addressed by many researchers in the field of mass spectrometry (MS)-based proteomics. The ability to identify thousands of proteins by shotgun-based strategies has made the mere idea of a global analysis of a particular protein modification seem reasonable. There has been much progress in the development of methods that make use of shotgun-based protein identification in the analysis of a wide variety of protein modifications, some of which will be discussed here. PMID- 15543968 TI - Surfing silica surfaces superciliously. AB - The present mini-review summarizes the experience gathered by our group in developing different classes of novel quaternarized heterocyclic compounds able to modulate and reverse the electroendoosmotic flow (EOF) in a most peculiar manner. The first class comprises mono-salt compounds, with the determinant omega iodoalkyl chains of different lengths (typically C4-C8), able to be adsorbed by silicas, at alkaline pH, and spontaneously alkylate ionised silanols, thus becoming covalently affixed to it. The second class is constituted by di-salt compounds, attached at the termini of an alkyl chain of variable lengths (here too, typically, C4-C8). This second class is unable to bind covalently silica surfaces, although, in thin-layer chromatography, it exhibits an extraordinary affinity for silica beads, contrary to the first one. On the basis of the strikingly different behaviour, structural rules are derived for the minimum requirements for general classes of amines to bind to silica walls and modify EOF. For compounds unable to bind covalently to the wall, the most important structural motif is two quaternary nitrogens spaced apart by a C4 chain: this seems to be the average distance (i.e., 0.8 nm) between two adjacent, ionized silanols for a snug fit. The other structural binding motif is the "hydrophobic decoration", i.e., the ratio of charged groups to alkyl residue in the various amines; amines with high levels of such alkane groups (i.e., with higher hydrophobicity), seem to bind more tenaciously to the wall, probably due to hydrophobic interaction not to the wall but among the amine derivatives themselves, when carpeting the silica. PMID- 15543969 TI - The role of liquid chromatography in proteomics. AB - Proteomics represents a significant challenge to separation scientists because of the diversity and complexity of proteins and peptides present in biological systems. Mass spectrometry as the central enabling technology in proteomics allows detection and identification of thousands of proteins and peptides in a single experiment. Liquid chromatography is recognized as an indispensable tool in proteomics research since it provides high-speed, high-resolution and high sensitivity separation of macromolecules. In addition, the unique features of chromatography enable the detection of low-abundance species such as post translationally modified proteins. Components such as phosphorylated proteins are often present in complex mixtures at vanishingly small concentrations. New chromatographic methods are needed to solve these analytical challenges, which are clearly formidable, but not insurmountable. This review covers recent advances in liquid chromatography, as it has impacted the area of proteomics. The future prospects for emerging chromatographic technologies such as monolithic capillary columns, high temperature chromatography and capillary electrochromatography are discussed. PMID- 15543970 TI - Sheathless electrospray ionization interfaces for capillary electrophoresis-mass spectrometric detection advantages and limitations. AB - A review of sheathless interfaces for capillary electrophoresis (CE)-mass spectrometry (MS) is presented. The review discusses the on-line CE-MS system requirements, advantages and weaknesses of current sheathless interface designs for CE-electrospray ionization MS, and comparison between sheath flow and sheathless interfaces. The advantages and limitations of three sheathless designs are discussed and commented upon, these include single-capillary, two-capillary and three-piece designs. PMID- 15543971 TI - Ionic boundaries in biological capillary electrophoresis. AB - Ion migration in electrophoresis always leads to the formation of ionic boundaries. While some types of the ionic boundaries can be formed intentionally, e.g., to improve separation efficiency or sensitivity of the analysis, other, naturally formed boundaries, may cause unexpected effects during the electrophoretic experiment. Some of the boundaries often go unnoticed in practice; however, many effects of the ionic boundaries formed by the sample matrix and background electrolyte components are frequently observed in capillary electrophoresis (CE). The interpretation of these effects may not be trivial and different explanations of the same phenomena may be found in the literature. This critical article attempts to review some of the important effects of the ionic boundaries observed or utilized during the CE of biologically important samples. PMID- 15543972 TI - Adsorption isotherms of the fullerenes C60 and C70 on a tetraphenylporphyrin bonded silica. AB - The single-component adsorption isotherms of the C60 (from 0 to 15 g/L) and C70 (from 0 to 8 g/L) buckminsterfullerenes on a tetraphenylporphyrin-bonded silica were acquired by frontal analysis, using a solution of toluene-1 methylnaphthalene (40:60, v/v) as the mobile phase. The best isotherm model derived from the fitting of these adsorption data was the bi-Langmuir model, a choice supported by the bimodal affinity energy distribution (AED) obtained for C60. The isotherm parameters derived from the inverse method (IM) of isotherm determination (by fitting calculated profiles to experimental overloaded band profiles of C60 and C70) are in very good agreement with those derived from the FA data. According to the isotherm parameters found by these three methods (FA, AED, IM), the tetraphenylporphyrin-bonded silica can adsorb 54 and 42 mmol/L of C60 and C70 fullerenes, respectively, a result that is consistent with the relative molecular size of these two compounds. The 20% lower surface accessibility for C70 is compensated by a three times higher equilibrium constant on the low-energy sites, giving a selectivity alpha(C70/C60) = 3.6. Large volumes (0.2, 0.8 and 1.7 mL) of mixtures of C60 (3.2 g/L) and C70 (1.3 g/L) were injected and their elution profiles compared to those calculated from the competitive bi-Langmuir model derived from the single-component isotherm data. A good agreement is obtained between calculated and experimental profiles, which supports the two-site adsorption mechanism derived from the single-component adsorption data. The measurements of the influence of the pressure on the retention of C60 and C70 demonstrate that the partial molar volumes of the two buckminsterfullerenes are 12 mL/mol larger in the stationary than in the mobile phase. PMID- 15543973 TI - Capillary liquid chromatographic determination of cellular flavins. AB - A capillary LC system was set up and optimized, in which a UV absorbance detector was used and a monolithic silica-ODS column as the separation column. Two on-line concentration techniques, namely, gradient elution mode and in-tube solid-phase ion-pair microextraction (SPIPME), were combined with the capillary LC system, which proved to be beneficial to enhance the concentration sensitivity by enabling the injection of large volumes of samples. The limits of detection at ppb levels for the flavins [riboflavin, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD)] were achieved using the two techniques. For in-tube SPIPME, a monolithic silica-ODS column was employed as the extraction column, on which FAD and FMN were retained b y interaction with an ion-pair reagent, tetrabutylammonium phosphate, resulting in greater than 110-fold enhancement in their concentration sensitivities relative to conventional injection method. The reproducibility and linearity of the two methods were investigated. The two methods were applied to analyze trace amounts of flavins in bacterial Escherichia coli cell extracts and recombinant flavoenzymes. PMID- 15543974 TI - Approach to the comprehensive analysis of glycoproteins isolated from human serum using a multi-lectin affinity column. AB - Glycosylation is one the most common post-translational modifications (PTM) and glycoproteins play fundamental roles in a diversity of biological processes. The development of an analytical approach to the study of variation of glycosylation patterns in serum samples has been hindered by the structural heterogeneity of this post-translational modification and the complexity of serum proteome. We have used the ability of different lectins to recognize specific glycosylation motifs to develop a specific affinity system that can achieve a comprehensive capture of serum glycoproteins. In a preliminary investigation, we evaluated the ability of five commonly used immobilized lectins to capture glycoproteins from human serum. SDS-PAGE analysis showed each lectin was able to enrich a subset of the serum glycoproteome and overlaps in lectin specificity were indeed observed. Based on these results and with the goal of studying the extent of the human serum glycoproteome, we then developed a multi-lectin affinity column containing Concanavalin A (Con A), Wheat germ and Jacalin lectin. The selection of lectins was also based on the known N-linked and O-linked glycan structures that are considered representative of the serum proteome. We then demonstrated that the capture of glycoproteins was specific, efficient and reproducible with this multi lectin column. The results obtained with this affinity step indicated that about 10% of human serum proteins are glycosylated (weight/weight) and, after removal of six high abundance proteins, including albumin, at least 50% of the remaining proteins were glycosylated. We then evaluated the use of this affinity column to monitor changes in the pattern of glycosylation in serum samples by a controlled, stepwise release of the captured proteins from the multi-lectin affinity column with specific displacers. PMID- 15543975 TI - Selective separations of peptides with sequence deletions, single amino acid polymorphisms, and/or epimeric centers using macrocyclic glycopeptide liquid chromatography stationary phases. AB - Separating closely related peptides (those differing by one or two amino acids or the chirality of a single amino acid) can be challenging using reversed-phase liquid chromatography (LC), ion-exchange LC, or using ion-pairing agents. Also, the mobile phases that give the best separations in these modes may not be electrospray ionization mass spectrometry (ESI-MS) compatible. Forty-two peptides from 11 peptide families were separated on three macrocyclic glycopeptide stationary phases in reverse-phase mode using ESI-MS-compatible mobile phases. The peptide classes studied were angiotensin, bradykinin, alpha-bag cell factor, beta,gamma-bag cell factor, beta-casomorphin, dynorphin, enkephalin, leucokinin, lutinizing hormone releasing hormone, neurotinsin, substance P, and vasopressin. High selectivity was observed for single amino acid substitutions (achiral and chiral) regardless of the position of the substitution in the sequence. Mobile phase optimization, its effect on peptide elution behavior, and chromatographic efficiency is also discussed. Using LC-ESI-MS, a 2 ng limit of detection was obtained, two orders of magnitude lower than the UV detection limit. PMID- 15543976 TI - Latex-functionalized monolithic columns for the separation of carbohydrates by micro anion-exchange chromatography. AB - A novel stationary phase for micro ion chromatography has been prepared by coating a porous poly(butyl methacrylate-co-ethylene dimethacrylate-co-2 acrylamido-2-methyl-1-propanesulfonic acid) monolith with quaternary amine functionalized latex particles via simple electrostatic binding. This stationary phase enabled the separation of saccharides in the mobile phase with a high-pH value consisting of aqueous ammonia solution in anion-exchange mode using evaporative light scattering for detection. Effects of both porous properties of the monolithic stationary phase and chromatographic conditions on the separation ability were studied. Under optimized conditions, an efficient separation of seven saccharides was achieved in less than 10 min. The stationary phase also enables the separation of saccharides obtained by the enzymatic hydrolysis of corn starch. PMID- 15543977 TI - Operational variables in high-performance liquid chromatography-electrospray ionization mass spectrometry of peptides and proteins using poly(styrene divinylbenzene) monoliths. AB - Capillary reversed-phase high-performance liquid chromatography (RP-HPLC) utilizing monolithic poly(styrene-divinylbenzene) columns was optimized for the coupling to electrospray ionization mass spectrometry (ESI-MS) by the application of various temperatures and mobile phase additives during peptide and protein analysis. Peak widths at half height improved significantly upon increasing the temperature and ranged from 2.0 to 5.4 s for peptide and protein separations at 70 degrees. Selectivity of peptide elution was significantly modulated by temperature, whereas the effect on proteins was only minor. A comparison of 0.10% formic acid (FA), 0.050% trifluoroacetic acid (TFA), and 0.050% heptafluorobutyric acid (HFBA) as mobile phase additives revealed that highest chromatographic efficiency but poorest mass spectrometric detectabilities were achieved with HFBA. Clusters of HFBA, water, and acetonitrile were observed in the mass spectra at m/z values >500. Although the signal-to-noise ratios for the individual peptides diverged considerably both in the selected ion chromatograms and extracted mass spectra, the average mass spectrometric detectabilities varied only by a factor of less than 1.7 measured with the different additives. Limits of detection for peptides with 500 nl sample volumes injected onto a 60 mm x 0.20 mm monolithic column were in the 0.2-13 fmol range. In the analysis of hydrophobic membrane proteins, HFBA enabled highest separation selectivity at the cost of lower mass spectral quality. The use of 0.050% TFA as mobile phase additive turned out to be the best compromise between chromatographic and mass spectrometric performance in the analysis of peptides and proteins by RP-HPLC-ESI MS using monolithic separation columns. PMID- 15543978 TI - Novel urea-linked cinchona-calixarene hybrid-type receptors for efficient chromatographic enantiomer separation of carbamate-protected cyclic amino acids. AB - Two novel diastereomeric cinchona-calixarene hybrid-type receptors (SOs) were synthesized by inter-linking 9-amino(9-deoxy)-quinine (AQN)/9-amino(9-deoxy) epiquinine (eAQN) and a calix[4]arene scaffold via an urea functional unit. Silica-supported chiral stationary phases (CSPs) derived from these SOs revealed, for N-protected amino acids, complementary chiral recognition profiles in terms of elution order and substrate specificity. The AQN-derived CSP showed narrow scoped enantioselectivity for open-chained amino acids bearing pi-acidic aromatic protecting groups, preferentially binding the (S)-enantiomers. In contrast, the eAQN congener exhibited broad chiral recognition capacity for open-chained as well as cyclic amino acids, and preferential binding of the (R)-enantiomers. Exceedingly strong retention due to nonenantioselective hydrophobic analyte calixarene interactions observed with hydro-organic mobile phases could be largely suppressed with organic mobile phases containing small amounts of acetic acid as acidic modifier. With the eAQN-calixarene hybrid-type CSP particularly high levels of enantioselectivity could be achieved for tert-butoxycarbonyl (Boc) , benzyloxycarbonyl (Z)- and fluorenylmethoxycarbonyl (Fmoc)-protected cyclic amino acids using chloroform as mobile phase, e.g. an enantioselectivty factor alpha >5.0 for Boc-proline. Increasing amounts of acetic acid compromised enantioselectivity, indicating the crucial contributions of hydrogen bonding to chiral recognition. Comparison of the performance characteristics of the urea linked eAQN-calixarene hybrid-type CSP with those of structurally closely related mutants provided evidence for the active involvement of the urea and calixarene units in the chiral recognition process. The urea linker motif was shown to contribute to analyte binding via multiple hydrogen bonding interactions, while the calixarene module is believed to support stereodiscrimination by enhancing the shape complementarity of the SO binding site. PMID- 15543979 TI - Use of two-dimensional liquid fractionation for separation of proteins from cell lysates without the presence of methionine oxidation. AB - A novel two-dimensional (2D) chromatographic method is developed to separate proteins from malignant breast cancer whole cell lysates. Protein mixtures are first separated according to their pIby chromatofocusing followed by an orthogonal non-porous reversed-phase separation. An important advantage of this 2D chromatographic method is that, unlike gel-based methods, it does not result in methionine oxidation. The lack of methionine oxidation during separation is demonstrated by the analysis of protein tryptic digests using matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) MS. Our novel 2D chromatographic method used in combination with on-target light-induced methionine oxidation provides a means for studying methionine-containing peptides. Methionine residues in peptide sequences are partially oxidized with light exposure. Neither the location nor the modification of methionine in the peptide sequence affects the oxidation. As a result, multiple peaks are observed in MALDI-TOF-MS spectra after light exposure. Sequence information derived from light-induced methionine can be applied to enhance the database search results obtained through peptide mass fingerprinting. PMID- 15543980 TI - Phenolic extraction of DNA from mammalian tissues and conversion to deoxyribonucleoside-5'-monophosphates devoid of ribonucleotides. AB - Towards a goal of detecting scaled-up DNA adducts as altered deoxynucleotides by mass spectrometry, we have set up a practical and general method for isolating DNA-derived deoxyribonucleoside-5'-monophosphates devoid of ribonucleotides starting with a 1 g sample of mammalian tissue. The method is practical because costs have been minimized, and it is general because it can be applied to a more difficult sample such as mouse skin or non-fresh calf liver. The procedure, consisting of a series of steps that were largely gleaned and tuned from prior literature, proceeds as follows: (1) homogenize the tissue in sodium dodecyl sulfate; (2) digest with ribonuclease A, ribonuclease TI, alpha-amylase and proteinase K; (3) partition between water and phenol; (4) precipitate the DNA with ethanol followed by redissolving and dialysis; and (5) digest with nuclease P1 and phosphodiesterase I followed by ultrafiltration and boric acid gel chromatography. The yellow to brown color of DNA from difficult tissues only persisted up to the ultrafiltration step. Apparently this DNA was contaminated with iron-containing proteins. Residual ribonucleotides were not observable (<0.1%) by HPLC in the final sample. Without boric acid gel chromatography, residual contamination by ribonucleotides was about 1% even when the DNA was purified before digestion by phenol partitioning followed by use of a Genomic Tip kit from Qiagen. PMID- 15543981 TI - Improved liquid chromatography-mass spectrometry performance in quantitative analysis using a nanosplitter interface. AB - Several pairs of analytes in plasma were investigated to demonstrate the successful utility of a novel interface in quantitative bioanalytical LC-MS and LC-MS/MS. Recently in our laboratory, an interface (the nanosplitter) was developed that allows the coupling of normal-bore liquid chromatography with microelectrospray mass spectrometry. The post-column concentric split minimizes turbulence and is shown to produce significant gains in the mass spectrometric signal. This configuration of the splitter allows sampling of the center portion of the parabolic HPLC plug, which maintains chromatographic integrity while producing high split ratios and effectively conserving nearly 99.9% of the sample. When utilizing a Finnigan mass spectrometer (with a heated capillary interface design), the signal gain with the nanosplitter ranged from 5 to 16 times the peak area obtained using the conventional interface without splitting. The linearity of the nanosplitter and conventional interface are shown to be comparable for all analytes tested. The nanosplitter was also fitted to a Sciex mass spectrometer and the results were compared to those from turbo ionspray. While in this case no significant signal improvement was observed, when normalized to the actual analyte mass introduced into the MS, the mass sensitivity was still increased 270-fold. The variations in signal gain utilizing the nanosplitter on instruments from different manufacturers reflect the inherent differences in the source designs while confirming the benefits of coupling high flow LC separations with low flow mass spectrometric detection. PMID- 15543982 TI - Monitoring the hydrophilicity/hydrophobicity of amino acid side-chains in the non polar and polar faces of amphipathic alpha-helices by reversed-phase and hydrophilic interaction/cation-exchange chromatography. AB - The ability to monitor precisely the hydrophobicity/hydrophilicity effects of amino acid substitutions in both the non-polar and polar faces of amphipathic alpha-helical peptides is critical in such areas as the rational de novo design of more effective antimicrobial peptides. The present study reports our initial results of employing the complementary separation modes of reversed-phase high performance liquid chromatography (RP-HPLC) and hydrophilic interaction/cation exchange chromatography (HILIC/CEX) to monitor the effect on apparent peptide hydrophilicity/hydrophobicity and amphipathicity of substituting single L- or D amino acids into the centre of the non-polar or polar faces of a 26-residue biologically active amphipathic alpha-helical peptide, V681. Our results clearly show that RP-HPLC and HILIC/CEX are best suited for resolving amphipathic peptides where substitutions are made in the non-polar and polar faces, respectively. Further, RP-HPLC and HILIC/CEX were demonstrated to be excellent monitors of hydrophilicity/hydrophobicity variations where amino acid substitutions were made in these respective faces. We believe these complementary high-performance modes offer excellent potential for rational design of novel amphipathic alpha-helical biologically active peptides. PMID- 15543983 TI - Single-molecule spectroscopy for molecular identification in capillary electrophoresis. AB - The electrophoretic mobilities of individual DNA molecules were determined by using fluorescence imaging. The average mobility agreed well with that obtained in capillary electrophoresis (CE). The signal-to-noise ratio (S/N) did not decrease in the presence of up to 8% plasma or 8% raw blood. Single-molecule detection was still possible in the presence of 50% raw blood. Single-molecule CE of two differently labeled molecules was carried out in the presence of a transmission grating. Even when the mobility difference is not sufficient because of low S/N, identification using different fluorescence wavelengths can be performed at >99% accuracy. So, when small differences in DNA sequence due to disease or mutation can lead to hybridization to labels with different dyes, the screening of the mutated DNA will be facilitated by online spectroscopy in addition to the electrophoretic information from CE. PMID- 15543984 TI - Influence of ignored and well-known zone distortions on the separation performance of proteins in capillary free zone electrophoresis with special reference to analysis in polyacrylamide-coated fused silica capillaries in various buffers. I. Theoretical studies. AB - Distortion of the starting zone upon its electrophoretic migration toward the detection window gives rise to both symmetrical zones caused by diffusion, sedimentation in the horizontal section of the capillary and the curvature of the capillary, and asymmetrical zones having their origin in Joule heating, sedimentation in the vertical section of the capillary, pH and conductivity differences between the sample zone and the surrounding buffer, solute adsorption onto the capillary wall, and association-dissociation of complexes between the analyte and a buffer constituent or between analytes. Interestingly and importantly a theoretical study shows that moderate pH and conductivity differences as well as adsorption and all of the above interactions when they are characterized by a fast on/off kinetics do not increase the zone broadening (or only slightly), because the sharpening of one boundary of the zone is about the same as the broadening of the other boundary. In addition the peak symmetry caused by a conductivity difference is in most experiments counteracted by a pH difference. The experimentally determined plate numbers in the absence of electroosmosis exceeded one million per meter in some experiments (Part II). These plate numbers are among the highest reported [Z. Zhao, A. Malik, M.L. Lee, Anal. Chem. 65 (1993) 2747; M. Gilges, K. Kleemiss, G. Schomburg, Anal. Chem. 66 (1994) 2038; H. Wan, M. Ohman, L.G. Blomberg, J. Chromatogr. A 924 (2001) 591 (plate numbers determined in the presence of electroosmosis may be higher, although the width of the zone in the capillary may be larger) [p. 680 in S. Hjerten, Electrophoresis 11 (1990) 665]). Capillary free zone electrophoresis is perhaps the only separation method, which, under optimum conditions, gives a plate number not far from the theoretical limit. A prerequisite for this high performance is that the polyacrylamide-coated capillary is washed with 2 M HCl between the runs and stored in water over night (Part II). The difference between the experimentally determined total variance and the sum of the calculated variances originating from the width of the starting zone, longitudinal diffusion, Joule heating, sedimentation in the vertical section of the capillary, curvature of the capillary (i.e., the sum of all other variances) was in our most successful experiments about 28% of the variance of diffusion. The zone broadening, 2sigma, caused by diffusion was estimated at 0.77 mm. The total zone width (2sigma) calculated from the experimentally determined plate number was as small as 1 mm when the migration distance was 40 cm. Accordingly, the only efficient way to reduce drastically the total zone width is to decrease the analysis time and, thereby, the diffusional broadening. An important finding was that the variance originating from the loops of the capillary is not always negligible in high-performance runs. Therefore, one should employ straight capillaries and avoid CE apparatus with cartridges that require a strong curvature of the capillary, common in most commercial instruments. Mathematical formulas have been derived for the sedimentation of the solute zone, the enrichment factor, and the migration time in experiments where the solute is dissolved in a dilute running buffer. This zone sharpening method gave very narrow starting zones (0.04-0.4 mm). However, upon high dilution of the buffer the enrichment becomes so strong that part of the sample zone probably sediments out of the capillary; the almost inevitable change in pH may decrease the mobility of the proteins and, thus, cause the enrichment factor to become still lower than expected. Diffusion of the protein in the very narrow starting zone (located close to the tip of the capillary) and sometimes the thermal expansion of the buffer in the capillary contributes to additional loss of protein in the enrichment step. In some buffers, the interaction between the protein and the buffer constituents is so slow that the peaks become broad. Therefore, different types of buffers should be tested when high resolution is required. The relation sigma2 (the variance of the interaction between a protein and the buffer constituents) = constant x u (the mobility) seems to be valid for all proteins in the applied sample, at least when they have similar molecular masses. To facilitate the understanding of the progress of a free zone electrophoresis experiment, we have discussed in simple terms how the concentrations of the background electrolytes become rearranged during a run and why the difference between the mobilities of the proteins and the mobilities of the background electrolyte determines whether a peak exhibits fronting or tailing. A theoretical analysis of zone broadening in capillary zone electrophoresis, chromatography, and electrochromatography indicates that electrochromatography in homogeneous gels might be the only chromatographic technique which can compete in performance with free electrophoresis. Using an equation, valid not only for electrophoresis, but also for chromatography and centrifugation, the mobility of a concentration boundary has been calculated for the first time and was, as expected, low. Equations based on the Kohlrausch regulating function do not permit such calculations. Another regulating function (the H function) and some of its characteristics are briefly discussed. The theoretical discussions in this paper and the experimental studies in Part II show that high-performance electrophoresis deserves its prefix when the runs are designed to give minimum zone broadening. Some guidelines are given to facilitate this optimization. The plate numbers are so high that the resolution cannot be increased by more than 30% even if they approach the theoretically maximum values. PMID- 15543985 TI - Influence of ignored and well-known zone distortions on the separation performance of proteins in capillary free zone electrophoresis with special reference to analysis in polyacrylamide-coated fused silica capillaries in various buffers. II. Experimental studies at acidic pH with on-line enrichment. AB - The separation of acidic and basic model proteins was studied in capillary free zone electrophoresis in a polyacrylamide-coated, electroosmosis-free capillary at pH below their isoelectric points (pI) using various buffers at pH 2.7-4.8 with UV detection at 200 nm. The separation performance was significantly dependent on the coating quality, which may even differ within the same batch of capillaries. In addition, a washing step with 2 M HCl and the storage of the capillary in distilled water was essential for the performance. For high efficiency and resolution the choice of buffer constituents was extremely important which is discussed in quantitative terms in Part I. The most promising buffers were ammonium acetate and ammonium hydroxyacetate at pH 4 (ionic strengths: 0.12 and 0.15 M, respectively) with plate numbers up to 1,700,000 plates/m, corresponding to a zone width (2sigma) of only 1 mm in a capillary with 40 cm effective length, when the injected samples were dissolved in a 10-fold diluted background electrolyte (BGE), a zone even narrower than those obtained in polyacrylamide gel electrophoresis, the characteristic feature of which is remarkably thin zones. In the experiment giving this plate number, the calculated variance for longitudinal diffusion was larger than all the other calculated variances (those for the width of the starting zone, Joule heating, sedimentation and the curvature of the capillary). Interestingly, the effect of capillary curvature was significant. In addition, the sum of all other imaginable variances (corresponding to various types of slow on/off kinetics and hyper-sharp peaks) was in the most successful experiments only 28-50% of the variance for longitudinal diffusion. One hundred- to two hundred-fold dilution of the BGE improved the detection limits and provided high precision in both migration times and peak areas with ammonium hydroxyacetate and ammonium acetate as background electrolytes. However, that high dilution increased the variance 140-400% for these buffers, respectively, at least partly due to conductivity or pH differences between the sample and buffer zones (hyper-sharp peaks). Sedimentation of the enriched sample, a factor that has not previously been treated theoretically or experimentally, was probably another reason for our finding that peak heights did not increase when the sample was dissolved in a buffer diluted more than 200-fold, although pH changes and in some cases thermal expansion in the capillary also may contribute. Loss of protein may occur at the ionic strength 0.01 and lower due to precipitation. Limits of detection were in the range 4-17 pmol of proteins with ammonium acetate as BGE. No indication of denaturation of proteins at pH 4 was observed. However, the separation performance at pH 3 was not satisfactory and loss of proteins was observed, possibly indicating such problems. The protein mobilities decreased unexpectedly from pH 4 to 3--a further indication of conformation changes. PMID- 15543986 TI - Determination of impurities in the drug 5-aminosalicylic acid by micellar electrokinetic capillary chromatography using an electrolyte pH that approaches the isoelectric point of the parent compound. AB - Process impurities found in the drug substance 5-aminosalicylic acid were determined by micellar electrokinetic capillary chromatography (MECC). In order to enhance the detection of trace impurities, the parent drug was dissolved to an unusually high concentration of 5 mg/mL. To reduce the dispersive processes of electromigration dispersion and anti-stacking, both of which occur at high solute concentration, the electrolyte pH was adjusted to be close to the apparent isoelectric point (pI) of the zwitterionic drug. In this fashion, the net charge on the solute should approach zero thereby minimizing the aforementioned sources of band-broadening. Two additional developments are reported. Short sodium hydroxide washes were used to optimize the MECC reproducibility. The elimination of anti-stacking permitted the use of peak heights to quantitate low level impurities with improved precision. The method was compared to the high performance liquid chromatographic (HPLC) method found in the United States Pharmacopoeia (USP). Both the MECC and HPLC methods were found to be similar with regard to migration-retention time reproducibility, peak area-height reproducibility, linearity and limit of quantitation. The MECC separation is 2x faster than HPLC. Both methods meet the system suitability requirements described in the USP. PMID- 15543987 TI - Determination of the amylose-amylopectin ratio of starches by iodine-affinity capillary electrophoresis. AB - This paper describes the application of capillary electrophoresis to separate and quantify the main polysaccharide components, amylose and amylopectin, present in starch samples. The separation is based on the well-known affinity of these compounds to iodine. The starch components could be effectively separated in less than 7 min using an uncoated fused-silica 'bubble cell' capillary. The proposed method has been applied for the quantitative determination of the soluble amylose content and the ratio amylose-amylopectin in commercial starches. It is shown that the present method is reliable, gives detection limits in the order of 0.1 mg mL(-1), is faster than other methodologies reported in the literature, and can be easily applied to the analysis of different starches. In spite of differences in solubility of amylopectin from different sources, a reasonable estimate of the amylose-amylopectin ratio could be made. Additionally, it was shown that the resulting profiles obtained after hydrolysis with isoamylase and alpha-amylase can provide information on the structure of the starches studied. PMID- 15543988 TI - Separation of fatty alcohol polyethoxylates by capillary electrophoresis through easy electroosmotic flow control with a quaternary diammonium salt. AB - An optimised protocol for the fast and reliable analysis of fatty alcohol polyethoxylates (FAEs), with resolution between the alkyl chain and ethylene oxide (EO) oligomers, is reported. Phthalic and maleic anhydrides were used to quickly form the hemiesters, which were separated by capillary electrophoresis. Effective reduction of the electroosmotic flow to very low values was achieved by previously rinsing the capillary with a quaternary diammonium salt, which remains strongly adsorbed to the silica wall. The phthalic hemiesters of a FAE mixture with 10 carbon atoms in the alkyl chain, and an average EO number of 6, were separated in a background electrolyte (BGE) containing 25 mM borate buffer of pH 9.0 in water. The same capillary pre-treatment was used to separate mixtures of the maleic hemiesters of hydrophobic FAEs, having 12-16 carbon atoms in the alkyl chain and an average EO number of 3. Full resolution of all the oligomers was achieved by using a finely tuned BGE in which a 35% borate buffer was mixed with 65% acetonitrile. PMID- 15543989 TI - Genotyping and haplotyping of the dopamine D4 receptor gene by capillary electrophoresis. AB - In this paper we report on simultaneous genotyping of adjacent polymorphisms (referred to as haplotyping) by combining double-tube allele-specific polymerase chain reaction, restriction fragment length polymorphism and capillary gel electrophoresis analysis of the resulting fragments. Direct molecular haplotyping is of particular importance in the case of double heterozygote samples, since in these instances the haplotype structure cannot be constructed based on genotype data. Our approach provided a powerful tool for coincidental genotype analysis of the 48 base pair (bp) variable number of tandem repeats of the third exon and haplotype investigation of the -616CG and -521CT single nucleotide polymorphisms of the dopamine D4 receptor (DRD4) gene. The linear polyacrylamide sieving matrix was optimized for the size range of the double-stranded DNA fragments of interest varying from 35 to 763 bp. We demonstrated that capillary gel electrophoresis in combination with laser induced fluorescence detection offers a sensitive and accurate tool for automated haplotyping in clinical settings. PMID- 15543990 TI - Capillary electrophoretic approach to screen for enzyme inhibitors in natural extracts. AB - This paper demonstrates development of electrophoretically mediated micro analysis (EMMA), for screening protein tyrosine phosphatase (PTP) inhibitors in natural extracts. It is demonstrated that capillary electrophoresis (CE) separation of the substrate and the product allows for using the assay in an on column format to monitor the reaction without typically used fluorogenic substrates. Michaelis-Menten kinetics parameters calculated based on the EMMA results (Km = 1.2-1.5 microM) were in a good agreement (Km = 1.0-1.5 microM) obtained using an off-line CE functional assay (CE FA). EMMA of PTP titrated with different concentrations of ligand demonstrated the peak-shift phenomenon normally seen in affinity capillary electrophoresis. This feature of EMMA gives an indication of the binding affinity of the ligand in addition to its functional activity, providing another dimension in characterization of the protein inhibitor interaction. It was demonstrated that simultaneous screening of the primary PTP target and a secondary, counter target (PTP-C) using the EMMA format can be used to prioritize hits based on their specificity. PMID- 15543991 TI - Capillary isoelectric focusing and affinity capillary electrophoresis approaches for the determination of binding constants for antibodies to the prion protein. AB - For the development of specific immunological assays, the binding of a specific antibody (Ab) to the target antigen (Ag) has to be relatively strong. In this study, we have utilized affinity capillary electrophoresis (ACE), a form of capillary zone electrophoresis (CZE) to determine the binding constant (Kb) of specific Abs against bovine serum albumin (BSA) and the healthy prion protein (PrPc), in buffer solutions at fixed pHs, approximating in vivo conditions. We have also utilized capillary isoelectric focusing (cIEF) to determine the complexity and recognition of the various isoforms of PrPc Abs towards their Ag, PrPc. Only ACE and CZE have been used to derive Kb values. The selected Abs for the prion protein can recognize both healthy and diseased states of the protein and are commercially available. The Kb values of PrPc Abs appear to be as strong as the anti-BSA (Ab to BSA) and other reported Kb values for proteins of similar size to PrPc. This appears to be one of the few reports on Kb values for any PrPc Abs, and their applications for in vitro immunoassays (e.g., enzyme-linked immunosorbent assays (ELISAs)). Such assays are being used to detect the infectious agent, PrPres, in brain and related matter/tissues. PMID- 15543992 TI - Chromatographic preconcentration coupled on-line to capillary electrophoresis via a tee-split interface. AB - Solid-phase extraction (SPE) and capillary electrophoresis (CE) are on-line coupled via a Tee-split interface, which provides hydrodynamic injection of the SPE eluate by flow splitting. The interface allows sample preconcentration independently from the CE separation and prevents sample matrix and washing solvents from entering the separation capillary. The effect of the Tee-split interface on the CE efficiency was examined using enkephalin peptides as model compounds. Most favorable plate numbers were obtained using a split ratio of 1:40. Breakthrough volume, desorption efficiency and elution volume for the C18 micro SPE column (5 mm x 0.5 mm i.d.) were found to be 750 microL, 65% and 1 microL, respectively. The performance of the complete system was demonstrated by the preconcentration and separation of an enkephalin mixture. Plate numbers up to 120,000 were obtained using a sample volume of 250 microL and a split ratio of 1:40. Enkephalin peak areas were linear (R2 = 0.996) over the 10-1000 ng/mL range. UV absorbance concentration limits of detection (SIN = 3) were about 5 ng/mL. For 250 microL injections of 100 ng/mL, the relative standard deviation (n = 5) of peak area was lower than 10%. PMID- 15543993 TI - Free flow electrophoresis coupled with liquid chromatography-mass spectrometry for a proteomic study of the human cell line (K562/CR3). AB - The requirement for prefractionation in proteomic analysis is linked to the challenge of performing such an analysis on complex biological samples and identifying low level components in the presence of numerous abundant housekeeping and structural proteins. The employment of a preliminary fractionation step results in a reduction of complexity in an individual fraction and permits more complete liquid chromatography/mass spectrometry (LC/MS) analysis. Free flow electrophoresis (FFE), a solution-based preparative isoelectric focusing technique, fractionates and enriches protein fractions according to their charge differences and is orthogonal in selectivity to the popular reversed phase high performance liquid chromatography (HPLC) fractionation step. In this paper, we explored the advantages of a combination of FFE and liquid chromatography/mass spectrometry to extend the dynamic range of a proteomic analysis of a complex cell lysate. In this study, the whole cell lysate of a chronic myelogeneous leukemia cell line, K562/CR3, was prefractionated by FFE into 96 fractions spanning pH 3-12. Of these, 35 fractions were digested with trypsin and then analyzed by LC/MS. Depending on the algorithm used for peptide assignment from MS/MS data, at least 319 proteins were identified through database searches. The results also suggested that pI could serve as an additional criterion besides peptide fragmentation pattern for protein identification, although in some cases, a pI shift might indicate post translational modification. In summary, this study demonstrated that free flow electrophoresis provided a useful prefractionation step for proteomic analysis and when combined with LC/MS allowed the identification of significant number of low level proteins in complex samples. PMID- 15543994 TI - Microscale protein expression profiling during disease evolvement. AB - Advances in technology, such as laser capture microdissection (LCM), have allowed for the specific sampling of cells within their natural functional micro environment. In model systems using LCM, we have studied the global protein expression profiles of airway epithelial cells during a response to allergen provocation. Bronchial epithelial cells were first identified and phenotyped histologically in snap frozen lung samples of experimentally sensitised mice. Consecutive thin sections of whole lung were then sampled using preparative LCM procedures. Lysates of the captured epithelium (7500 shots) or whole lung were prepared for two-dimensional gel electrophoretic separation and 1400 protein spots were annotated by image analysis. Protein identities were established by matching peptide masses detected using matrix-assisted laser desorption ionization time-of-flight MS as well as electrospray ionization MS-MS sequencing. Using the Mascot database of protein/peptide identities high significance scores in terms of sequence coverage (range 22-70%) and number of peptides (range 7-22 peptides/protein) were obtained for approximately 500 proteins, with examples listed in Table 1. In quantitative terms, the LCM procedure allows the statistical sampling of singular populations of cells distributed throughout tissues and organs. The absolute number of cells required for "entry level" measurements of protein profiles will vary over an order of magnitude depending on the physical size and frequency of the cells being studied within each biological compartment as well as the dynamic range of the proteins being measured, and the absolute limits of detection within the technologies being employed. PMID- 15543995 TI - Effect of solvent composition on signal intensity in liquid chromatography-matrix assisted laser desorption ionization experiments. AB - The use of reversed phase liquid chromatography for the preparation of complex peptide mixtures for analysis by matrix assisted laser desorption ionization mass spectrometry has led to the observation of the critical importance of the matrix/analyte formulation in regards to the percent organic solvent in the mixture. This paper outlines the study using liquid chromatography to systematically vary the acetonitrile concentration in the formulation used for MALDI spot preparation to examine the impact the parameter has on analyte signal intensity. The results show that for five of six peptides tested across a wide mass range a formulation of approximately 75% acetonitrile is optimal for average MALDI signal intensity as determined on both time-of-flight and quadropole mass spectrometers. Examination of the individual spots shows that the organic solvent content in formulation significantly affects parameters such as crystal density and morphology. PMID- 15543996 TI - Development of an analytical reversed-phase high-performance liquid chromatography-electrospray ionization mass spectrometry method for characterization of recombinant antibodies. AB - Analytical characterization of monoclonal antibodies has been hindered by the lack of appropriate chromatographic methods to be used in conjunction with high resolution MS. Current methodologies for standard RP-HPLC are incompatible with antibodies due to irreproducibility, low recovery, short column lifetimes, and poor resolution of degradation products. An analytical RP-HPLC-MS method was developed for monitoring and characterizing intact IgG1antibodies. Key parameters required for improved chromatographic resolution included long alkyl chains of the stationary phase (Zorbax SB300 C8), column temperatures elevated to 65-70 degrees C and combination of trifluoroacetic acid and heptafluorobutyric acid ion pairing agents. RP chromatographic separation of degradation species and C terminal lysine variants along with the characterization of glycosylation profile by mass spectrometry demonstrates the capability of this method for whole antibody analysis. PMID- 15543997 TI - Sleep disorders: an overview. AB - Although sleep disorders medicine is a relatively young discipline, understanding of the diagnosis, pathophysiology, and treatment of sleep disorders is evolving at a rapid pace. This overview discusses the history of the development of sleep disorders medicine, tracing changes in the diagnostic classification of sleep disorders as well as the role of polysomnography in diagnosis. This evolution is most evident for insomnia, one of the major sleep disturbances. The accumulation of epidemiologic data on the prevalence and temporal course of insomnia and emerging information regarding its pathophysiology derived from laboratory assessments have led to the development of new therapeutic approaches for primary insomnia and insomnia associated with medical and psychiatric disorders. PMID- 15543998 TI - Chronic insomnia: current issues. AB - Insomnia is a common problem in the general population and has a higher prevalence in persons with medical and psychiatric disorders. Although insomnia is most often transient, occurring as a result of identifiable stressors, a substantial portion of insomnia cases involve persistent sleep difficulty. This chronic form of insomnia may be associated with a wide range of adverse consequences. An understanding of the characteristics and causes of this disorder and the available therapeutic strategies will promote more effective identification and treatment of patients with chronic insomnia. PMID- 15543999 TI - [Prevalence of transmission of methicillin resistant Staphylococcus in home care patients]. AB - Methicilline-resistant Staphylococcus aureus (MRSA) is among the most important causes of hospital-acquired infections. The aim of the present study was to investigate the prevalence of MRSA carriage among home care patients as well as among the health care personnel taking care of those patients. 93 patients and 30 staff members of the team of a home care sector in the western part of Luxembourg were screened for MRSA carriage and factors associated with carriage were investigated among the patients. PMID- 15544000 TI - [Cerebral apoplexy--importance of diagnosis and therapy in acute stroke]. AB - Every day occur about 4 strokes in the Grand-Duchy of Luxemburg. Stroke is the third cause of death and the first cause of acquired handicap. Stroke constitutes a major individual and social problem. Stroke and TIA are neurological emergency conditions that warrant immediate hospitalisation on a stroke unit, where a dedicated equipe of neurologists, nurses, speech and occupational therapists, physiotherapists and social workers in collaboration with other medical and surgical specialties provide up-to-date diagnostic and therapeutic measures. These include the diagnostic work-up of stroke aetiology and the vascular status, monitoring to ensure the maintenance of homeostasis, thrombolysis and revascularisation, antithrombotic drug treatment relying on individual stroke pathophysiology, treatment of risk factors and early rehabilitation. By treatment on a stroke unit, death and disability can be reduced by 27%, in the case of intravenous thrombolysis even by 45%. PMID- 15544001 TI - [Evaluation of the national epidemiological program of breast cancer screening in the Grand Duchy of Luxembourg, 1999-2002]. AB - In 1992 a national breast cancer screening programme was launched in Luxembourg with following characteristics; invitation at 2-yearly interval, 2 views, double reading for women aged between 50 and 64 and since 2001, those aged 65-69 are as well included. From the beginning on, one of the main concern was to respect a rigorous methodology, in accordance with the recommendations of the European Commission. This paper presents the early performance indicators of the years 1999-2002 of the "Programme Mammographie". Almost all histopathological results were obtained, because only one national pathology laboratory exists in Luxembourg, where also the national tumor's register is located. In ten years, a total of 91.432 mammograms were performed. The results of the early performance indicators are in agreement with the European Recommendations. A 36% detection rate of small size cancers, < or = 10 mm, and since 2000, more then 70% of node negative cancers are observed among women aged 50-64 attending the Programme. The cancer rate with a good prognostic factor for the patients is high. Despite good results, the rate of interval cancers is still too high, and two conclusions can be drown: the women's participation at screening has to be steady in time without trespassing a two years intervals, and after the detection of small cancer a high quality assessment should be followed by an adequate treatment, leading to a reduction of mortality and also to a decrease of morbidity by treatment. PMID- 15544002 TI - [Professor Francis Naville (1883 - 1968), His role in the inquiry on the massacre in Katyn]. AB - On April 13th 1943, the German radio announced that a common grave containing the corpses of Polish officers was found in the forest of Katyn, near Smolensk. Further enquiries showed that the dead bodies were those of officers imprisoned by the Russians in autumn 1939 and about who nobody had news since Springtime 1940. The CICR refused to deal with this problem without the agreement of the Soviets. So the Germans called together a committee of international experts to examine the grave in Katyn. Prof. Francis Naville, director of the medico-legal Institute of the University of Geneva, was the only expert really neutral in that commission. He had the merit to prove quite clearly that these criminal executions were ordered by Stalin. After the Second World War, Prof. Naville was criticised for having accepted to participate to the mission to Katyn by Jean Vincent, a deputy of the Swiss labour party (communist party) who claimed that the massacre of Katyn had been done by the Germans. Prof. Naville got no support from the CICR, who "did not want to know" who was responsible in order to avoid diplomatic complications with the Soviet Union. Only in 1989 the discovery of Prof. Naville was accepted and confirmed by the Soviet authorities. PMID- 15544003 TI - [The impact of crisis intervention on the management of patients with personality disorder]. AB - The aim of this study was to investigate in psychiatric emergency unit at Hospital Center of Luxembourg the impact of crisis intervention on the management of patients with personality disorder. There were included in this study all the patients with the diagnosis of personality disorder (axis II, criteria DSM IV), even the reason for emergency consultation was related to a disorder diagnosed on the axis I (criteria DSM IV). Data were retrospectively collected to compare patients examined in the psychiatric emergency unit during five months, from 1st September 2001 to 31st January 2002, with those examined from 1st September 2002 to 31st January 2003 and able to benefit from a crisis intervention. The results show a significant reduction of voluntary hospitalizations and a significant increase of the number of patients with more than five outpatient consultations, after the introduction of crisis interventions. The theoretical frame of our psychotherapeutic interventions is briefly exposed. The results of this study emphasize the importance of crisis interventions in psychiatry for improving the management of patients with a personality disorder. PMID- 15544004 TI - [Radiotherapy at the Grand Duchy of Luxembourg]. AB - The Centre Francois Baclesse was created as National Radiotherapy Center in 2000. Oncology Radiotherapy has been developed in Luxembourg in parallel with existing medical and surgical oncology departments. Cancer is a Public Health problem in Luxembourg. More than 1800 new cases of cancer are diagnosed every year, out of wich 70 % will need radiotherapy at the time of the evolution of the disease. Technical platform and Human Resources available at this time are able to deliver high quality treatment. The activity reached a plateau in 2002 with almost 800 patients treated per year, whereas the theorical need for radiation oncology treatment is 1200 patients per year in Luxembourg. A large architectural project will add 2 new linear accelerators and will be finished by 2007-2008. Multidisciplinary oncological network has been developed together with cancer treatment units of 6 luxembourgish hospitals. Clinical Research is one of the missions of the Center. After the achievement of its extension, the Centre Francois Baclesse will be able to cover the need of the Regional, luxembourgish and cross-boarder patients pool, in line with an European approach in treating cancer. PMID- 15544005 TI - [Early detection of synchronous thyroid carcinoma and malignant melanoma by fluorodeoxyglucose positron emission tomography]. AB - Functional tumor imaging using Fluorodeoxyglucose positron emission tomography (FDG-PET) is a new method in clinical oncology. The 18 FDG, is a glucose analog that accumulates in cells in proportion to the rate of glucose metabolism, and increased carbohydrate metabolism has been recognized as a feature of malignant cells versus normal cells. In addition, it permits the detection of metastases or synchronous tumours not discovered by anatomic imaging. Although detection of the primary site of disease is usually accomplished well with conventional techniques, the performance of FDG-PET may be useful to determine tumours that are not clinically evident. The authors describe a case of early detection of synchronous thyroid carcinoma by FDG-PET in a young patient opereted on for a malignant melanoma on his arm. PMID- 15544006 TI - Primitive abscess of the great epiploon: clinical case and literature review. AB - BACKGROUND: The primitive abscess of the great epiploon often poses a pre operative diagnosis problem. The clinical picture associates an abdominal pain and an inflammatory syndrome. METHODS: The diagnosis is essentially based on the imaging data and the exploratory laparoscopy. The treatment consists in performing a drainage of the abscess and an epiploic resection "on demand" preserving healthy tissues, as well as giving antibiotics. RESULTS: The patient has been treated successfully by coelioscopy and a partial epiploic resection was performed with uncomplicated post-operative outcome. CONCLUSIONS: The primitive abscess of the great epiploon remains a rare pathology of uncertain etiology. The coelioscopy is the best method to rectify or to comfirm pre-operative diagnosis and to perform treatment avoiding the inconveniences of the laparotomy. PMID- 15544007 TI - Review of laparoscopic and open colorectal surgery in the "Zitha" Hospital (Luxembourg) in the year 2002. AB - BACKGROUND: A review and outcome of laparoscopic colorectal surgery in the visceral surgery unit in the Zitha Hospital for the year 2002. METHODS: All patients, presenting with benign or malignant disease of the colon or rectum were evaluated. Depending on the tumour size, a laparoscopic approach is advocated whenever possible. Operation and patient specific factors (age, duration of the procedure and duration of hospital stay, diagnosis of malignant tumour versus benign disease, conversion and complication rate) are presented. Patients who had to be converted are classified under the open surgery group. RESULTS: A total of 103 laparoscopic or open procedures on the colon/rectum were performed between the 1st of January 2002 and the 31st of December 2002. 74% were treated by a laparoscopic and 26% by an open approach. 10 out of 103 patients did not undergo a colonic resection but instead an emergency colostomy/ ileostomy or oversewing of a bowel leak, in 8 cases performed laparoscopically. 44 patients have been operated for a benign disease (36 laparoscopic and 8 open procedures) and 59 patients have been treated for a malignant disease (40 laparoscopic and 19 open procedures). The average operating time for the laparoscopic resection was 160 minutes in comparison to 182 minutes for the open resection. The conversion rate (inclusive of non-resection procedures) was 5% for a benign disease and 11% for colorectal cancer. The minor complication rate was 16% in laparoscopic surgery and 30% in open surgery. We saw two major complications in both the open and laparoscopic groups (4%). Reoperation was necessary in one patient out of 103. The average postoperative hospital stay was 7.78 days for the laparoscopic and 16.6 days for the open group. The hospital mortality was 2.9%. CONCLUSION: The laparoscopic colorectal resection is a safe and beneficial procedure for the patient when used in experienced hands. PMID- 15544008 TI - [Puncture-lavage-infiltration of calcifying tendinitis of the rotator cuff]. AB - Calcifying tendinitis of rotator cuff is a common and evolutive pathology of the shoulder. This article supports the efficiency of imaging-guided needle puncture trituration and lavage in rotator cuff calcifications after failure of medical treatment. Overall efficiency of this economical ambulatory treatment ranges in the literature from 49 to 100 % and no complication is noted. In our opinion this technique should be used before arthroscopic or surgical removal of the calcifying tendinitis. PMID- 15544009 TI - Beta3 integrins: major therapeutic targets of the near future. AB - Integrins are major cell adhesion receptors which assume two important functions: first they act as anchoring molecules allowing firm cellular attachment to the extracellular matrix, and second they work as signalling receptors being able to transduce signals in both directions (outside-in and inside-out) through the plasma membrane. Their biological importance is determined by their involvement in many physiological phenomena. Furthermore, their implication in various diseases and their accessibility to drugs make them interesting therapeutic targets. PMID- 15544010 TI - [Luxembourg meeting for clinical and experimental virology. Luxembourg, 2709.02]. PMID- 15544011 TI - [50th anniversary of the Luxembourg Society of sports medicine]. PMID- 15544012 TI - SARS revisited: the challenge of controlling emerging infectious diseases at the local, regional, federal, and global levels. PMID- 15544013 TI - Sudden unexplained death caused by cardiac ryanodine receptor (RyR2) mutations. PMID- 15544014 TI - Eight-month prospective study of 14 patients with hospital-acquired severe acute respiratory syndrome. AB - OBJECTIVE: To define the clinical characteristics and clinical course of hospital acquired severe acute respiratory syndrome (SARS). PATIENTS AND METHODS: This 8 month prospective study of 14 patients with hospital-acquired SARS in Taipei, Taiwan, was conducted from April through December 2003. RESULTS: The most common presenting symptoms in our 14 patients with hospital-acquired SARS were fever, dyspnea, dizziness, malaise, diarrhea, dry cough, muscle pain, and chills. Lymphopenia and elevated serum levels of lactate dehydrogenase (LDH) and C reactive protein (CRP) were the most common Initial laboratory findings. Initial chest radiographs revealed various pattern abnormalities and normal results. Five of the 14 patients required mechanical ventilation. The need for mechanical ventilation was associated with bilateral lung involvement on the initial chest radiograph and higher peak levels of LDH and CRP. Clinical severity of disease varied from mild to severe. At 8 months after disease onset, patients with mild or moderate SARS had normal findings or only focal fibrosis on chest high resolution computed tomography. However, bilateral fibrotic changes remained in the 4 patients who had recovered from severe SARS, 1 of whom had mild restrictive ventilatory impairment. One patient with severe SARS died; she was elderly and had other comorbidities. Five additional patients had reduced diffusing capacity. CONCLUSION: The clinical picture of our patients presenting with hospital acquired SARS revealed atypical pneumonia associated with lymphopenia, elevated serum levels of LDH, rapid clinical deterioration, and lack of response to empirical antibiotic therapy. Substantially elevated levels of LDH and CRP correlated with severe illness requiring mechanical ventilatory support. In those receiving mechanical ventilation, pulmonary function was only mildly reduced at 6 to 8 months after acute illness, consistent with the natural history of acute respiratory distress syndrome due to other causes. PMID- 15544015 TI - Targeted mutational analysis of the RyR2-encoded cardiac ryanodine receptor in sudden unexplained death: a molecular autopsy of 49 medical examiner/coroner's cases. AB - OBJECTIVE: To perform a molecular autopsy of the RyR2-encoded cardiac ryanodine receptor/calcium release channel in medical examiner/coroner's cases of sudden unexplained death (SUD). METHODS: From September 1998 to March 2004, 49 cases of SUD were referred by medical examiners/coroners to the Sudden Death Genomics Laboratory at the Mayo Clinic in Rochester, Minn, for a cardiac channel molecular autopsy. Mutational analysis of 18 exons of RyR2 implicated previously in the pathogenesis of catecholaminergic polymorphic ventricular tachycardia (CPVT) was performed on genomic DNA using polymerase chain reaction, denaturing high performance liquid chromatography, and direct DNA sequencing. RESULTS: This cohort of 49 cases of SUD included 30 males, 13 with a family history of syncope, cardiac arrest, or sudden cardiac death (mean +/- SD age at death, 14.2 +/- 10.9 years). Six distinct RyR2 missense mutations (3 novel) were discovered in 7 cases (14%, 6 males, mean +/- SD age at death, 13.6 +/- 11.2 years) of SUD. The activities at the time of SUD were exertion (3), emotion (1), and unknown (3). The mutations, R420W, S2246L, N4097S, E4146K, T4158P, and R4497C, involved nonconservative amino acid substitutions in highly conserved residues across species and were not seen in 400 reference alleles. CONCLUSIONS: This study represents the first molecular autopsy of RyR2 in medical examiner-referred cases of SUD. A targeted analysis of only 18 of the 105 protein-encoding exons of the cardiac ryanodine receptor/calcium release channel revealed potential CPVT1 causing RyR2 mutations in 1 of every 7 cases of SUD. These findings suggest that postmortem genetic testing of RyR2 should be considered as a part of the comprehensive medicolegal autopsy investigation of a SUD case and that this potentially heritable and often elusive arrhythmia syndrome be scrutinized carefully in family members of those who experience SUD. PMID- 15544016 TI - Outcome of patients with end-stage renal disease admitted to the intensive care unit. AB - OBJECTIVES: To describe the clinical course of patients with end-stage renal disease (ESRD) admitted to the intensive care unit (ICU) and to compare the performance of Acute Physiology and Chronic Health Evaluation (APACHE) III and Sequential Organ Failure Assessment (SOFA) in predicting their outcome. PATIENTS AND METHODS: This retrospective cohort study consisted of patients with ESRD admitted to 3 ICUs between January 1, 1997, and November 30, 2002. Data on demographics, APACHE III score, SOFA score, development of sepsis and organ failure, use of mechanical ventilation, and mortality were collected. RESULTS: Of the 476 patients with ESRD who underwent dialysis during the study period, 93 (20%) required admission to the ICU. The most common ICU admission diagnosis was gastrointestinal bleeding. The first day median (Interquartile range) APACHE III score, SOFA score, and APACHE III predicted hospital mortality rate were 64 (47 79), 6 (5-8), and 12.9% (4.2%-30.8%), respectively. The observed ICU, hospital, and 30-day mortality rates were 9%, 16%, and 22%, respectively. Nonrenal organ failure developed in 48 patients (52%) and sepsis in 15 patients (16%). Mechanical ventilation was required In 26 patients (28%). The area under the receiver operating characteristic curve for the first-day APACHE III probability of hospital death in predicting 30-day mortality was 0.78 (95% confidence interval, 0.68-0.86) compared with 0.66 (95% confidence interval, 0.55-0.76) for the SOFA score (P = .16). CONCLUSIONS: The observed hospital mortality of patients with ESRD admitted to the ICU is relatively low. There is no statistically significant difference in the performance of APACHE III and SOFA prognostic models in discriminating between 30-day survivors and nonsurvivors. PMID- 15544017 TI - Cardiopulmonary resuscitation in critically ill neurologic-neurosurgical patients. AB - OBJECTIVES: To establish the rate of successful cardiopulmonary resuscitation (CPR) and to study outcome predictors in patients who experienced in-hospital cardiac arrest after being admitted to the neurologic-neurosurgical intensive care unit (ICU) with a primary neurologic diagnosis. PATIENTS AND METHODS: We identified patients admitted to the neurologic-neurosurgical ICU between 1994 and 2001 who experienced in-hospital cardiac arrest and received CPR. Functional outcome was assessed using the modified Rankin scale. RESULTS: During the study period, 38 consecutive patients experienced in-hospital cardiac arrest and received CPR. The median age of the patients was 65 years (range, 16-81 years), and the mean interval from admission to CPR was 12 days (range, 3 hours to 47 days). Acute intracranial disease was present in 32 patients (84%). Twenty-one patients (55%) were in the ICU at the time of the cardiac arrest; cardiac arrests in the wards occurred at a mean interval of 9 days (range, 1-45 days) after ICU discharge. Cardiopulmonary resuscitation achieved return of spontaneous circulation in 23 patients (61%). Seven patients (18%) were discharged from the hospital, 5 of whom later achieved a modified Rankin scale score of 2 or lower. Cardiac arrest after a deteriorating clinical course resulted in uniformly fatal outcomes. Duration of CPR shorter than 5 minutes and CPR in the ICU were associated with survival and good functional recovery. CONCLUSIONS: Cardiopulmonary resuscitation is a worthwhile procedure in severely ill neurologic-neurosurgical patients, regardless of the patient's age. However, the outcome after CPR appears much worse in patients with a prior deteriorating clinical course. PMID- 15544018 TI - Safety of electroconvulsive therapy in patients receiving long-term warfarin therapy. AB - OBJECTIVE: To investigate the safety of electroconvulsive therapy (ECT) in patients receiving long-term warfarin therapy. PATIENTS AND METHODS: Retrospective data were reviewed for 35 consecutively hospitalized patients who received long-term warfarin therapy and ECT at the Mayo Clinic in Rochester, Minn, between January 1, 1994, and December 31, 2001. RESULTS: A total of 300 ECT treatments were administered to the 35 patients. Of 284 ECT treatments for which data were available, no ECT-related complications due to anticoagulation occurred despite increases in blood pressure and pulse rate. One patient experienced ventricular tachycardia, resulting in transfer to a cardiology service for temporary monitoring. No other serious ECT-related adverse effects were noted. The rate of intertreatment delirium was similar to that reported in other studies. CONCLUSIONS: Electroconvulsive therapy in patients receiving long-term warfarin therapy appears to be safe. Although no major adverse effects were identified in our case series, additional prospective evaluation is warranted. PMID- 15544019 TI - Development and validation of the ORACLE score to predict risk of osteoporosis. AB - OBJECTIVE: To develop and validate a composite index, the Osteoporosis Risk Assessment by Composite Linear Estimate (ORACLE), that includes risk factors and ultrasonometric outcomes to screen for osteoporosis. SUBJECTS AND METHODS: Two cohorts of postmenopausal women aged 45 years and older participated in the development (n = 407) and the validation (n = 202) of ORACLE. Their bone mineral density was determined by dual energy x-ray absorptiometry and quantitative ultrasonometry (QUS), and their historical and clinical risk factors were assessed (January to June 2003). Logistic regression analysis was used to select significant predictors of bone mineral density, whereas receiver operating characteristic (ROC) analysis was used to assess the discriminatory performance of ORACLE. RESULTS: The final logistic regression model retained 4 biometric or historical variables and 1 ultrasonometric outcome. The ROC areas under the curves (AUCs) for ORACLE were 84% for the prediction of osteoporosis and 78% for low bone mass. A sensitivity of 90% corresponded to a specificity of 50% for identification of women at risk of developing osteoporosis. The corresponding positive and negative predictive values were 86% and 54%, respectively, in the development cohort. In the validation cohort, the AUCs for identification of osteoporosis and low bone mass were 81% and 76% for ORACLE, 69% and 64% for QUS T score, 71% and 68% for QUS ultrasonometric bone profile index, and 76% and 75% for Osteoporosis Self-assessment Tool, respectively. ORACLE had the best discriminatory performance in identifying osteoporosis compared with the other approaches (P < .05). CONCLUSION: ORACLE exhibited the highest discriminatory properties compared with ultrasonography alone or other previously validated risk indices. It may be helpful to enhance the predictive value of QUS. PMID- 15544020 TI - Autopsy results after surgery for non-small cell lung cancer. AB - OBJECTIVE: To determine the percentage of metastatic and unexpected residual lung cancer at autopsy in patients considered for curative resection of non-small cell lung cancer during a time when computed tomography was available as a preoperative staging tool. MATERIAL AND METHODS: Clinical data and surgical and autopsy slides of all patients who underwent curative resection of nonsmall cell lung cancer at the Mayo Clinic in Rochester, Minn, between 1985 and 1999 and who underwent autopsy within 30 days of surgery were reviewed retrospectively for the presence of residual or metastatic disease. RESULTS: The study group consisted of 25 men and 7 women, with a mean age of 70 years. A pulmonary metastasis was identified at surgery in 1 patient (3%). Metastases were found in an additional 5 patients (16%) at autopsy, 1 of whom had 2 sites involved. These sites included the liver in 2 and lung, epicardium, adrenal gland, and kidney in 1 each. The average diameter of metastases was 1.6 cm. No factor studied was found to be significantly associated with the presence of unrecognized metastatic disease at autopsy. CONCLUSION: The advent of computed tomography as a staging tool has decreased the percentage of patients with undiagnosed metastatic disease at surgery; however, preoperative understaging in lung cancer remains a problem. PMID- 15544021 TI - Persons with dementia who become lost in the community: a case study, current research, and recommendations. AB - Providing care for persons with dementia presents several serious challenges. Among these is the possibility that a person with dementia will become lost in the community and face the risk of injury or death. Caregivers often cite anxiety about this possibility as a reason for placing their loved ones in a professional care setting. Our case study and review of research show that all persons with dementia are at risk, regardless of age, past behavior, and sex. Thus, health care providers have an important role to play in educating caregivers about this risk and assisting them in preventing or responding to such situations. In addition, health care providers should ensure that their respective professional care facilities have appropriate measures in place to prevent and respond to these eventualities. Finally, health care providers can educate local law enforcement personnel about dementia in general and more specifically about research-based strategies for searching for persons with dementia who have become lost in the community. PMID- 15544022 TI - alpha1-Adrenoceptor subtype selectivity and lower urinary tract symptoms. AB - Benign prostatic hyperplasia is a common cause of urinary flow obstruction in aging men and may lead to lower urinary tract symptoms (LUTS). Benign prostatic hyperplasia has 2 physiological components: a static component related to increased prostate size and a dynamic component related to increased prostate smooth muscle tone. alpha1-Adrenoceptors (alpha1ARs) maintain prostate smooth muscle tone; hence, alpha1-antagonists (blockers) relax prostate smooth muscle and decrease urethral resistance, ultimately leading to relief of LUTS. This review focuses on alpha1AR subtypes and their location in lower urinary tract tissues involved in LUTS (prostate, bladder, spinal cord); it also summarizes major clinical trials published to date on the efficacy of alpha1AR blockers for LUTS. Benefits and adverse effects of clinically available alpha1AR antagonists are reviewed, followed by recent information on interactions between alpha1AR subtype antagonists and type 5 phosphodiesterase inhibitors used for impotence. alpha1-Adrenoceptor antagonists have become the mainstay of therapy for LUTS; knowledge about specific alpha1AR subtypes should facilitate rational choice of alpha1AR blocker therapy by clinicians. PMID- 15544023 TI - 76-year-old woman with chest pain. PMID- 15544024 TI - Initial evaluation and management of infertility by the primary care physician. AB - Infertility is a common condition seen in primary care practices. Infertility is defined as 1 year of unprotected intercourse during which a pregnancy is not achieved. in the United States, 15% to 20% of all couples are infertile, with higher rates seen in older couples. The causes of infertility include abnormalities of any portion of the male or female reproductive system. The female partner usually presents initially for an infertilty problem, often in the context of an annual well-women examination. The primary care physician who provides such preventive care can initiate the diagnostic evaluation and can treat some causes of infertility. PMID- 15544025 TI - Life-threatening serotonin syndrome in a patient with chronic heart failure and CYP2D6*1/*5. AB - We report a case of serotonin syndrome that occurred in a patient with chronic heart failure associated with a panic disorder. The 39-year-old Japanese man had been treated with paroxetine at 20 mg/d for 1 1/2 years. He presented with rhabdomyolysis, renal failure, fulminant liver failure, cardiac conduction disturbance, and disseminated intravascular coagulation, as well as conventional symptoms of serotonin syndrome including alterations in cognition (disorientation, confusion) and behavior (restlessness), autonomic nervous system dysfunction (fever, shivering), and abnormal neuromuscular activity (ataxia, hyperreflexia, myoclonus). All medications prescribed before hospital admission were discontinued. After 24 hours of continuous venovenous hemofiltration, diuresis resumed and renal and liver function improved rapidly. Disorientation, restlessness, hyperreflexia, and myoclonus abated slowly over the next 72 hours. The patient's anxiety subsided more slowly, and he recovered completely 1 week later. The plasma concentration of paroxetine was elevated far above the upper limit of the therapeutic range. The patient had cytochrome P-450 (CYP) 2D6*1/*5, a heterozygosity of an inactivated allele of CYP2D6, which metabolizes paroxetine. The patient was determined to be an intermediate metabolizer who was potentially vulnerable to paroxetine, a major inhibitor of CYP2D6. Heart failure is often accompanied by psychiatric disorders. A wide range of drugs commonly prescribed for these conditions, including beta-blockers, antiarrhythmics, and antidepressants, are metabolized by CYP2D6. Genetic screening for CYP2D6 in patients with these conditions may prevent life-threatening drug intoxication. PMID- 15544026 TI - Endocarditis caused by Coccidioides species. AB - Endocarditis caused by Coccidioides species has been reported rarely. Herein, we report 2 such cases and summarize 4 published reports. Coccidioidal endocarditis was found in patients who had prolonged or disseminated infection. Vegetations were identified on mitral or aortic valves or valvular rings, and embolic phenomena were observed. Diagnosis was hindered by uniformly negative results from blood cultures. The patients had a wide range of serologic titers for Coccidioides species (1:1-1:2048). The infection was fatal in 4 of the 6 patients whose cases we reviewed. We conclude that coccidioidal endocarditis is an uncommon manifestation of Coccidioides infection that connotes a poor prognosis. PMID- 15544027 TI - Pulmonary embolization of 2-octyl cyanoacrylate after endoscopic injection therapy for gastric variceal bleeding. AB - N-butyl-2-cyanoacrylate, a tissue adhesive that polymerizes on contact with weak bases such as blood, is being used widely outside the United States to obliterate gastric varices. Embolization of this material can occur via portosystemic shunts. We report a case of pulmonary embolization of 2-octyl cyanoacrylate (an analogue of N-butyl-2-cyanoacrylate) that occurred after endoscopic injection therapy for gastric variceal bleeding. Cyanoacrylate embolism is difficult to diagnose with computed tomographic angiography because radiopaque emboli are masked by the contrast material. It is important to distinguish these emboli from conventional thromboemboli because "glue emboli" require only symptomatic treatment. Clinicians should have a high index of suspicion for embolism in the setting of tachycardia, chest pain, or hypoxia after a patient undergoes endoscopic injection therapy with cyanoacrylate glue for gastric variceal bleeding. The radiologist should be alerted so that the appropriate radiographic settings are used to make the diagnosis. PMID- 15544028 TI - Management of acute ischemic stroke. AB - The treatment of acute ischemic stroke has evolved from observation and the passage of time dictating outcome to an approach that emphasizes time from ictus, rapid response, and a dedicated treatment team. We review the treatment of acute ischemic stroke from the prehospital setting, to the emergency department, to the inpatient hospital setting. We discuss the importance of prehospital assessment and treatment, including the use of elements of the neurologic examination, recognition of symptoms that can mimic those of acute ischemic stroke, and rapid transport of patients who are potential candidates for thrombolytic therapy to hospitals with that capability. Coordinated management of acute ischemic stroke in the emergency department is critical as well, beginning with non-contrast enhanced computed tomography of the brain. The advantages of a multidisciplinary dedicated stroke team are discussed, as are thrombolytic therapy and other inpatient treatment options. Finally, we cover evolving management strategies, treatments, and tools that could improve patient outcomes. PMID- 15544029 TI - Delusional parasitosis. PMID- 15544030 TI - Growth factor activation in myocardial vascularization: therapeutic implications. AB - A rapid growth of the coronary vasculature occurs during prenatal and early postnatal periods as precursor cells from the epi- and sub-epicardium differentiate, migrate and form vascular structures (vasculogenesis) which then fuse, branch and in some cases recruit cells to form three tunics (angiogenesis). These processes are tightly controlled by temporally and spatially expressed growth factors which are stimulated by metabolic and mechanical factors. The process of angiogenesis in the myocardium is not limited to developmental periods of life, but may occur when the heart is challenged by enhanced loading conditions or during hypoxia or ischemia. This review focuses on the activation of growth factors by metabolic and mechanical stimuli in the developing heart and in the adult heart undergoing adaptive responses. Experimental studies support the hypotheses that both metabolic (hypoxia) and mechanical (stretch) factors serve as powerful stimuli for the up-regulation of growth factors which facilitate angiogenesis and arteriogenesis. Both hypoxia and stretch are powerful inducers of VEGF and its receptors, and provide for paracrine and autocrine signaling. In addition to the VEGF family, bFGF and angiopoietins play major roles in myocardial vascularization. Sufficient evidence supports the hypothesis that mechanical (e.g., bradycardia) and metabolic (e.g., thyroxine analogs) may provide effective non-invasive angiogenic therapies for the ischemic and post infarcted heart. PMID- 15544031 TI - Angiogenic signal during cardiac repair. AB - Despite significant advances in myocardial revascularization and reperfusion, coronary artery disease and subsequently myocardial infarction, are the leading causes of morbidity and mortality in the United States. Strategies which improve the myocardial substrate during and following a myocardial infarction-such as the regrowth of functional blood vessels to the ischemic myocardium would be of great clinical importance. This review article attempts to address this important clinical issue through identifying potential signalling mechanisms by various mode of preconditioning that cause angiogenesis. PMID- 15544032 TI - Nitric oxide signaling during myocardial angiogenesis. AB - Ischemic heart disease develops as a consequence of coronary atherosclerotic lesion formation. Coronary collateral vessels and microvascular angiogenesis develop as an adaptive response to myocardial ischemia, which ameliorates the function of the damaged heart. Angiogenesis, the formation of new blood vessels from pre-existing vascular bed, is of paramount importance in the maintenance of vascular integrity both in the repair process of damaged tissue and in the formation of collateral vessels in response to tissue ischemia. Angiogenesis is modulated by a multitude of cytokines/chemokines and growth factors. In this regard, angiogenesis cannot be viewed as a single process. It is likely that different mediators are involved in different phases of angiogenesis. Vascular endothelial cells (ECs) produce nitric oxide (NO), an endothelium-derived labile molecule, which maintains vascular homeostasis and thereby prevents vascular atherosclerotic changes. In patients with ischemic heart disease, the release of endothelium-derived NO is decreased, which plays an important role in the atherosclerotic disease progression. In recent years, endothelium-derived NO has been shown to modulate angiogenesis in vitro and in vivo. In this review, we summarize recent progress in the field of the NO-mediated regulation of postnatal angiogenesis, particularly in response to myocardial ischemia. PMID- 15544033 TI - Myocardial angiogenesis. AB - Coronary angiogenesis and collateral growth are chronic adaptations to myocardial ischemia, which are aimed at restoring coronary blood flow and salvaging myocardium in an ischemic region. Although we have assumed that myriad numbers of growth factors are involving in this adaptation, details in the underlying mechanisms, i.e., number of angiogenic factors, angiostatic factors, their receptors/signaling cascades, interactions/crosstalk among the signaling pathways and receptors, and the time course of expression/function of a particular factor or pathway during the successful adaptation are still unclear; they are, probably, harmonized like a symphony. Although there is as of yet no consensus about the mechanisms and causal factors for these cononary adaptations to ischemia, recent evidence strongly suggests that a balance between growth factors and growth inhibitors is critical. In this review we introduce vascular endothelial growth factor, angiopoietins, and angiostatin, as factors playing pivotal roles in coronary angiogenesis and collateral growth. PMID- 15544034 TI - Verification of coronary angiogenesis by quantitative morphology. AB - The angiogenic response may be reliably evaluated only by the methods of quantitative morphology. These methods may appear deceivingly simple but they contain several possible pitfalls. This review presents major principles of proper methodology for determination of tissue vascularization using quantitative morphology. Description of appropriate preparation of the tissue is followed by a survey of methods available for visualization of the vascular structures, by the description of principles for proper sampling and measurements and, finally, by a section on how to present and interpret the results. PMID- 15544035 TI - Complexity in the vascular permeability factor/vascular endothelial growth factor (VPF/VEGF)-receptors signaling. AB - The adult vasculature results from a network of vessels that is originally derived in the embryo by vasculogenesis, a process whereby vessels are formed de novo from endothelial cell (EC) precursors, known as angioblasts. During vasculogenesis, angioblasts proliferate and come together to form an initial network of vessels, also known as the primary capillary plexus. Sprouting and branching of new vessels from the preexisting vessels in the process of angiogenesis remodel the capillary plexus. Normal angiogenesis, a well-balanced process, is important in the embryo to promote primary vascular tree as well as an adequate vasculature from developing organs. On the other hand, pathological angiogenesis which frequently occurs in tumors, rheumatoid arthritis, diabetic retinopathy and other circumstances can induce their own blood supply from the preexisting vasculature in a route that is close to normal angiogenesis. Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) is perhaps the most important of pro-angiogenic cytokine because of its ability to regulate most of the steps in the angiogenic cascade. The main goal of this review article is to discuss the complex nature of the mode of action of VPF/VEGF on vascular endothelium. To this end, we conclude that more research needs to be done for completely understanding the VPF/VEGF biology with relation to angiogenesis. PMID- 15544036 TI - Therapeutic myocardial angiogenesis with vascular endothelial growth factors. AB - Emerging evidence has shown that administration of angiogenic growth factors, either as recombinant protein or by gene transfer, can augment tissue perfusion through neovascularization in animal models of myocardial and hindlimb ischemia. Many cytokines have angiogenic activity; one of those that have been best studied in animal models and clinical trials is vascular endothelial growth factor (VEGF). VEGF has been known to be a key regulator of physiologic and pathologic angiogenesis associated with tumor. Recently the effect of VEGF is not restricted to the direct angiogenic effect in vivo but includes mobilization of bone-marrow derived endothelial progenitor cells and augmentation of postnatal vasculogenesis in situ. Clinical trials of therapeutic angiogenesis with VEGF in patients with end-stage coronary artery disease have shown increases in exercise time and reductions in anginal symptoms and have provided objective evidence of improved perfusion and left ventricular function. Larger scale placebo-controlled trials with recombinant protein (rhVEGF165) have been limited to intracoronary and intravenous administration and have shown favorable trends in exercise time and angina frequency. Small-scale, placebo-controlled, randomized clinical trials of gene transfer (phVEGF-2) via thoracotomy or percutaneous intramyocardial delivery demonstrated significant improvement of both subjective symptoms and objective measures of myocardial ischemia. Both therapeutic modalities appear to be safe and well tolerated. Further studies are required to determine the optimal dose, formulation, route of administration, and combinations of growth factors and the utility of adjunctive endothelial progenitor cell or other stem cell supplementation, to provide safe and effective therapeutic myocardial neovascularization. PMID- 15544039 TI - Integrative signaling in angiogenesis. AB - Angiogenesis is a complex process requiring integration of multiple signals in order to achieve successful development of the new vasculature. While individual activities of numerous growth factors are well understood, the integration of their signaling at the cellular and tissue level is just beginning to be appreciated. This review focuses on these two process using vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF2) as examples. PMID- 15544037 TI - Therapeutic myocardial angiogenesis: past, present and future. AB - One of the main goals in the treatment of myocardial ischemia is the development of effective therapy for angiogenesis and neovascularization. The first evidence demonstrating alleviation of myocardial ischemia and increased number of collateral blood vessels was reported in the early 90s following intra-coronary administration of basic fibroblast growth factor protein in canine. This study established the ground for extensive investigations to demonstrate the use of other angiogenic growth factor proteins, genes administered directly or incorporated in viruses, and more recently, endothelial progenitor stem cells (embryonic and adults). The positive results observed in animals failed, in most cases, to repeat themselves in clinical-trials in human patients. Therefore, additional experiments are warranted to allow full understanding of the mechanism underlying new blood vessel formation before further clinical studies are undertaken. This review will explore the milestones of angiogenic investigations and their clinical application. PMID- 15544040 TI - Non-classical mechanisms of heart repair. AB - Vasculogenesis was long considered to be a mechanism of blood vessel formation limited to the embryo, and not used in adult vascular repair. Similarly, myocardial proliferation was generally thought to cease in the adult. Yet there is an extensive older literature suggesting that blood might be a source of adult angioblasts and somewhat more recent data indicate that cardiomyocytes may proliferate. Only recently has the significance of this literature been recognized and the findings more closely investigated. It is now clear that bone marrow-cells move into the blood as angioblasts and contribute to vascular repair and neovascularization in the adult. Bone marrow cells can also move into the damaged myocardium, proliferate, and repair the damaged heart. This review will discuss data relevant to these non-classical mechanisms of heart repair, principally vascular repair, and some of the scientific history that led to modern thinking. Though the literature in this field is replete with examples of conclusions not warranted by the data, despite oft-exaggerated claims, the data do show that these non-classical mechanisms can contribute to cardiovascular maintenance and repair. What remains to be deciphered is the physiological importance of the mechanisms. PMID- 15544038 TI - Reactive oxygen species as mediators of angiogenesis signaling: role of NAD(P)H oxidase. AB - Angiogenesis, a process of new blood vessel growth, contributes to various pathophysiologies such as cancer, diabetic retinopathy and atherosclerosis. Accumulating evidence suggests that cardiovascular diseases are associated with increased oxidative stress in blood vessels. Reactive oxygen species (ROS) such as superoxide and H2O2 cause blood vessels to thicken, produce inflammation in the vessel wall, and thus are regarded as "risk factors" for vascular disease, whereas ROS also act as signaling molecules in many aspects of growth factor mediated physiological responses. Recent reports suggest that ROS play an important role in angiogenesis; however, its underlying molecular mechanisms remain unknown. Vascular endothelial growth factor (VEGF) induces angiogenesis by stimulating endothelial cell (EC) proliferation and migration primarily through the receptor tyrosine kinase VEGF receptor2 (Flk1/KDR). VEGF binding initiates tyrosine phosphorylation of KDR, which results in activation of downstream signaling enzymes including ERK1/2, Akt and eNOS, which contribute to angiogenic related responses in EC. Importantly, the major source of ROS in EC is a NAD(P)H oxidase and EC express all the components of phagocytic NAD(P)H oxidase including gp91phox, p22phox, p47phox, p67phox and the small G protein Rac1. We have recently demonstrated that ROS derived from NAD(P)H oxidase are critically important for VEGF signaling in vitro and angiogenesis in vivo. Furthermore, a peptide hormone, angiotensin II, a major stimulus for vascular NAD(P)H oxidase, also plays an important role in angiogenesis. Because EC migration and proliferation are primary features of the process of myocardial angiogenesis, we would like to focus on the recent progress that has been made in the emerging area of NAD(P)H oxidase-derived ROS-dependent signaling in ECs, and discuss the possible roles in angiogenesis. Understanding these mechanisms may provide insight into the components of NAD(P)H oxidase as potential therapeutic targets for treatment of angiogenesis-dependent diseases such as cancer and atherosclerosis and for promoting myocardial angiogenesis in ischemic heart diseases. PMID- 15544041 TI - Protein-, gene-, and cell-based therapeutic angiogenesis for the treatment of myocardial ischemia. AB - Therapeutic angiogenesis aims at restoring perfusion to chronically ischemic myocardial territories by using growth factors or cells, without intervening on the epicardial coronary arteries. Despite angiogenesis having received considerable scientific attention over the last decade, it has not yet been shown to provide clinical benefit and is still reserved for patients who have failed conventional therapies. Nevertheless, angiogenesis is a very potent physiologic process involved in the growth and development of every animal and human, and it is likely that its use for therapeutic purposes, once its underlying mechanistic basis is better understood, will one day become an important modality for patients with CAD and other types of organ ischemia. This review summarizes current knowledge in therapeutic angiogenesis research. PMID- 15544043 TI - Angiogenic strategy for human ischemic heart disease: brief overview. AB - In the Western World ischemic coronary disease is the leading cause of morbidity and mortality. Therapeutic approaches mostly aim to restore flow to a localized segment by angioplasty or bypass surgery. Therapeutic angiogenesis and or arteriogenesis describes a strategy where blood vessel formation is induced for the purposes of treating and/or preventing ischemic disease. At present, at least 17 clinical trials of myocardial angiogenesis have been presented involving over 900 patients. Therapeutic angiogenesis makes use of the administration of angiogenic growth factor protein or gene to promote the development of endogenous collateral vessels in ischemic myocardium. Most recently, interest has grown in the potential angiogenesis effects of cell therapy--such as autologous bone marrow cells or cultured stem cells--and there are now several groups initiating phase I/II trials in this area. PMID- 15544042 TI - Neovascularization derived from cell transplantation in ischemic myocardium. AB - Myocardial ischemia triggers a limited angiogenic response, part of the remodeling process that is insufficient to avoid further functional impairment. Several strategies have been evaluated to regenerate myocardial vascularization after ischemic injury such as transmyocardial laser revascularization and gene therapy. Attention has recently been focused on the potential of cell therapy to induce angiogenesis. Enhancing myocardial neovascularization is a major goal of myocardial cell transplantation because it would provide patients, who cannot undergo conventional revascularization, with an alternative therapy. Additionally, neovascularization would provide the implanted cells with adequate microenvironment to enhance survival and function. This short review gives an overview of the effect of various cell transplantation strategies on myocardial neovascularization. It suggests that in order to optimize myocardial neovascularization induced by cell therapy, future experiments should focus on the contribution of exogenous and endogenous stem cells to new vessels formation, and on the identification of the molecular pathways involved in the process. PMID- 15544044 TI - Therapeutic angiogenesis for severe ischemic heart diseases by autologous bone marrow cells transplantation. AB - Conventional therapies for severe ischemic heart disease are limited in applicability. While several angiogenesis researches have shown novel efficacy, safety and feasibility for clinical use, recently we have started the clinical trial of a sole cell therapy using autologous bone marrow mononuclear cells transplantation targeted into ischemic hibernating myocardium. Here, we review the background of bone marrow cell research and introduce therapeutic angiogenesis for severe ischemic heart disease by autologous bone marrow cells transplantation. PMID- 15544045 TI - Current clinical perspectives on myocardial angiogenesis. AB - Currently accepted modalities of treatment for atherosclerotic coronary artery disease (CAD) include pharmacological therapy, and revascularization with either bypass surgery or percutaneous coronary intervention (PCI). Similarly, conventional treatment of congestive heart failure (HF) is limited to medical therapy, temporary assist devices and in a select few, cardiac transplantation. A significant subset of patients with severe symptomatic CAD and end stage HF is not eligible for these traditional methods of treatment. In spite of maximal medical and revascularization therapy, these patients may not get adequate symptomatic relief. After a decade of investigations, gene therapy is emerging as a promising therapeutic option for this group of patients. This review discusses myocardial angiogenesis as a therapeutic modality in these patients including therapeutic angiogenesis with growth factors and cell transplantation. PMID- 15544047 TI - Expression of matrix metalloproteinase activity in idiopathic dilated cardiomyopathy: a marker of cardiac dilatation. AB - BACKGROUND: Idiopathic dilated cardiomyopathy (DCM), ventricular systolic dysfunction and chamber dilatation are accompanied by architectural remodeling, wall thinning and cardiac myocyte slippage. Recent work has demonstrated an association between collagen degradation and an increased expression of matrix metalloproteinases (MMPs). Accordingly, we have sought to correlate (a) collagen degradation with MMP elevations and, (b) assay the neutralizing potential of a known inhibitor of MMP, tetracycline on MMPs in DCM. METHODS: Assessment of LV volume and shape by 2-D echocardiography was performed. Light microscopic assessment of histopathology in picrosirius red stained biopsy samples of 11 DCM patients and six post-transplant patients was performed. Zymographic estimation of MMP activity and influence of tetracycline on MMP activity was assessed. RESULTS: Small amount of interstitial collagen was noted in the control group, whereas in the DCM hearts, chamber dilatation was associated with areas of scanty myocyte necrosis, islands of excess collagen, and focal areas of absent or scanty collagen with intact myocytes. In cardiomyopathic tissue, collagenase activity was markedly elevated at 63% compared with 8% in post-transplant tissue. Tetracycline at a concentration of 285+/-10 microM (IC50) inhibited collagenase activity by 50% in cardiomyopathic tissue. CONCLUSIONS: Areas of focal interstitial collagen accumulation were accompanied by collagen fiber lysis and increased collagenase activity in dilated cardiomyopathy. This enhanced collagenolytic activity found in endomyocardial biopsy tissue was inhibited by tetracycline. The non-antibiotic property of tetracycline may be of potential value in the prevention of ventricular dilatation in idiopathic dilated cardiomyopathy. PMID- 15544046 TI - Gene transfer of CuZn superoxide dismutase enhances the synthesis of vascular endothelial growth factor. AB - Nitric oxide (NO) and reactive oxygen species (ROS) are emerging as important regulators of angiogenesis. NO enhances VEGF synthesis in several cell types and is required for execution of VEGF angiogenic effect in endothelial cells. Similarly, hydrogen peroxide induces VEGF synthesis and recent studies indicate the involvement of ROS in signaling downstream of VEGF stimulation. VEGF synthesis can not only be enhanced by gene transfer of VEGF but also by overexpression of NO synthase genes. Here, we examined the possibility of augmentation of VEGF production by gene transfer of copper/zinc superoxide dismutase (CuZnSOD, SOD1). Overexpression of human SOD1 in mouse NIH 3T3 fibroblasts increased SOD activity, enhanced intracellular generation of H2O2 and significantly stimulated VEGF production as determined by increase in VEGF promoter activity, VEGF mRNA expression and VEGF protein synthesis. The stimulatory effect on VEGF synthesis induced by SOD1 gene transfer was reverted by overexpression of human catalase. The effect of H2O2 produced by engineered cells is mediated by activation of hypoxia-inducible factor response element (HRE) as well as Sp1 recognition site of VEGF promoter. This data suggest the feasibility of stimulation of angiogenesis by overexpression of SOD1. PMID- 15544048 TI - Expression of endothelial nitric oxide synthase in the vascular wall during arteriogenesis. AB - Nitric oxide (NO) has been demonstrated to play an important role in angiogenesis, and also to be involved in collateral vessel growth. The expression of endothelial NO synthase (eNOS) is moderated partly by blood flow-induced mechanical factors, i.e., shear stress. The purpose of this study was to evaluate how the expression of eNOS correlates with the development of collateral vessels in dog heart, induced by chronic occlusion of the left circumflex artery. Immunoconfocal microscopy using an antibody against eNOS was used to detect expression of eNOS in different stages of arteriogenesis. Collateral vessels were classified into normal, growing and mature vessels by using the cytoskeleton marker desmin. Expression of the growth factors bFGF and metallproteinase-2 (MMP 2) was also examined. The data show that in normal arteriolar vessels, expression of eNOS is very low, but in growing collateral vessel there is a 6.2-fold increase, which, however, returned to normal levels in mature collateral vessels. The expression of eNOS was localized only in endothelium, either in normal or growing vessels. bFGF was very weakly stained in normal vessels, but highly expressed in growing collateral vessels. MMP-2 was strongly stained in neointima, but very weak in endothelium. In addition, we also examined expression of iNOS because iNOS may be induced in vessel injury or in disease states, but it was not detected in either normal or growing collateral vessels. Our findings indicate that the expression pattern of eNOS is closely associated with the development of collateral vessels, suggesting that eNOS plays an important role in arteriogenesis. PMID- 15544049 TI - Remodeling of the vascular tunica media is essential for development of collateral vessels in the canine heart. AB - Previous studies have shown that neointima formation and adventitial remodeling play an important role in the enlargement of collateral vessels (CVs) during coronary arteriogenesis in the dog heart. In this study, we investigated the importance of remodeling of the tunica media in the same model. Basal membrane (BM), contractile and cytoskeletal components of smooth muscle cells (SMCs) were studied in growth of coronary CVs induced by chronic occlusion of the left circumflex (LCX) coronary artery by routine histology, electron microscopy (EM), and immunoconfocal microscopy using antibodies against alpha-smooth actin (alpha SM actin), calponin, desmin, and laminin. In addition, matrix metalloproteinase-2 (MMP-2) and tissue inhibitor-1 of matrix metalloproteinase (TIMP-1) were investigated. The data showed that (1) in normal small arteries (NVs) laminin formed a network in which SMCs were encaged; alpha-SM actin, calponin and desmin were evenly expressed in SMCs; (2) in early (2 weeks) growing CVs the laminin network was disrupted, desmin was significantly reduced in SMCs, but alpha-SM actin and calponin still highly expressed; (3) in actively (6 weeks) growing CVs laminin was still weak in the tunica media (TM), but without network-like structure. Desmin was further reduced in SMCs of TM, whereas alpha-SM actin and calponin showed little changes, although they were significantly decreased in intimal SMCs; (4) in mature CVs, the network-like structure was re-formed, and alpha-SM actin, calponin, and desmin were all similar to that in normal vessels; (5) histology for BM confirmed laminin staining; (6) EM revealed that in NVs the SMCs contained abundant contractile filaments and were surrounded by a layer of BM whereas in growing CVs, BM structure was not observed, but the SMCs in the media still contained many myofilaments; (7) MMP-2 was highly expressed in the media of early growing vessels, but decreased in TM of actively growing vessels where TIMP-1 expression was high. In conclusion, our data revealed features of TM of growing CVs. Disruption and degradation of BM facilitate SMC proliferation, and together with reduction of desmin and fragmentation of the internal elastic lamina enable the vascular wall to expand and enlarge when blood pressure and shear stress increase. MMP2 may be an important player in regulating SMC phenotype, proliferation, migration and maintaining integrity of the vascular wall through governing proteolysis during arteriogenesis. PMID- 15544050 TI - Synthesis and in vitro evaluation of new azaphenylalanine derivatives as serine protease inhibitors. AB - New inhibitors of serine proteases with azaphenylalanine scaffold were synthesized and their activity was evaluated in vitro. We studied the effect of different substituents in the part of a molecule that binds in the distal pocket of the thrombin active site. Modifications generally led to decreased activity, however two derivatives are promising lead compounds as new thrombin and dual thrombin-factor Xa inhibitors. PMID- 15544051 TI - 2-(Arylpropionylamino)- and 2-(arylacryloylamino)benzophenones: farnesyltransferase inhibition and antimalarial activity. AB - Structural variation of the 2-acylamino moiety of some benzophenone farnesyltransferase inhibitors led to the para-trifluoromethylphenylpropionyl derivative with relatively low farnesyltransferase inhibition but considerable antimalarial activity and no cytotoxicity. PMID- 15544052 TI - Synthesis and pharmacological investigation of some novel 2-methyl-3-(substituted methylamino)-(3H)-quinazolin-4-ones as histamine H1-receptor blockers. AB - A series of 2-methyl-3-(substituted methylamino)-(3H)-quinazolin-4-ones were synthesized from 3-amino-2-methyl-(3H)-quinazolin-4-one. Their structures were confirmed by spectral data (IR, NMR and MS) and the purity was ascertained by microanalysis. When tested for H1-receptor blocking activity on isolated guinea pig ileum all the test compounds inhibited histamine induced contraction whereas compound 5 (IC50 0.22 x 10(3) ng/ml) was found to be four times more potent than chlorpheniramine maleate (IC50 1 x 10(3) ng/ml) and it showed lesser sedation (8%) than the standard (32%). PMID- 15544053 TI - Improved synthesis, antibacterial activity and potential carcinogenicity of 5 amino-1 ,2,4-thiadiazol-3(2H)-one. AB - This paper reports on the preparation of 5-amino-1,2,4-thiadiazol-3(2H)-one, a sulfur-containing analogue of cytosine with the -CH=CH- group between the positions 5 and 6 of the pyrimidine ring replaced by the divalent sulfur (-S-). Improved procedures for the preparation of thiobiuret, some of its methyl derivatives and 5-amino-1,2,4-thiadiazol-3(2H)-one are documented. Thiobiuret and its N-methyl derivatives were obtained by addition of hydrogen sulfide to the respective 1-cyanoureas. Subsequent oxidation of thiobiuret with hydrogen peroxide in alkaline medium produced 5-amino-1,2,4-thiadiazol-3(2H)-one. This substance was traced back converted to the starting thiobiuret by reaction with cysteine hydrochloride. Alkaline degradation of thiadiazol led to the formation of 1-cyanourea isolated as its silver salt. An investigation of the thiadiazol biological activities has shown that it inhibits the growth of E. coil by 10% at 8.5 microM concentrations, but exhibited no cytostatic activity in L1210, HeLa S3 and HL-60 cell lines. Potential carcinogenicity of the prepared compounds was determined by a DC polarographic method. While the values of the parameter of carcinogenicity for all intermediates indicate only marginal carcinogenic potential, the value of the parameter of carcinogenicity for the thiadiazole indicates possible carcinogenicity of this compound. PMID- 15544054 TI - [Medium sized rings from alprenolol and oxprenolol]. AB - The N-allyl derivatives 2 formed from the beta-blockers 1, were acetylated to give the esters 5. The 1,5-benzoxazacycloundecine 6a and the 1,10,5 benzodioxazacyclododecine 7a were isolated by ring closing metathesis (RCM) of 5a and 5b, respectively, using Grubbs catalyst in the presence of titanium tetraisopropanolate. Alkaline hydrolysis yielded the alcohols 6b and 7b. PMID- 15544055 TI - HPLC profiling and quantification of active principles in leaves of Hedera helix L. AB - Ivy (Hedera helix L., Araliaceae), is an evergreen medicinal and ornamental plant. Depending on leaf polymorphism different shaped ivy leaves were extracted and subsequently analyzed by reversed-phase high performance liquid chromatography (RP-HPLC). Quantitative determination of its most prominent saponins hederacoside C (1) and alpha-hederin (2) from different ivy leaf extracts were detected, validated and optimized for quick profiling. The linearity of response, repeatability and reproducibility of the applied RP-HPLC method are reported. PMID- 15544056 TI - In vitro analysis of rifampicin and its effect on quality control tests of rifampicin containing dosage forms. AB - The chemical stability of rifampicin both in solid state and various media has widely been investigated. While rifampicin is appreciably stable in solid-state, its decomposition rate is very high in acidic as well as in alkaline medium and a variety of decomposition products were identified. The literature reports on highly variable rifampicin decomposition in acidic medium. Hence, the objective of this investigation was to study possible reasons responsible for this variability. For this purpose, filter validation and correlation between rifampicin and its degradation products were developed to account for the loss of rifampicin in acidic media. For analysis of rifampicin with or without the presence of isoniazid, a simple and accurate method was developed using high performance chromatography recommended in FDC monographs of the United States Pharmacopoeia. Using the equations developed in this investigation, the amount of rifampicin degraded in the acidic media was calculated from the area under curve of the degradation products. Further, it was proved that in a dissolution study, the colorimetric method of analysis recommended in the United States Pharmacopoeia provides accurate results regarding rifampicin release. Filter type, time of injection as well as interpretation of data are important factors that affect analysis results of rifampicin in in vitro studies and quality control. PMID- 15544057 TI - In vitro evaluation of fixed dose combination tablets of anti-tuberculosis drugs after real time storage at ambient conditions. AB - Rifampicin exhibits variable bioavailability from solid oral dosage forms and this problem is more apparent when it is formulated as fixed dose combination (FDC) in presence of other first-line anti-tuberculosis drugs. To determine the cause of variable bioavailability, the aging effect on physical and chemical performance of rifampicin from FDC formulations after real time storage at the ambient conditions was investigated. For this purpose, six FDC formulations from different manufacturers were stored at ambient conditions (20-35 degrees C, with no control of humidity) in the final packing for a period of 16-38 months and its in vitro quality control tests for rifampicin were compared with the initial performance of these tablets. None of the formulations have shown significant weight gain/loss and the assay values were within the pharmacopeial limits when evaluated by a stability indicating method. Further storage had no effect on physical performance of FDC tablets as indicated by unaltered dissolution profiles. Formulation reevaluation after real time storage at the ambient conditions for 16-38 months indicated that rifampicin containing FDC formulations are stable throughout its shelf life and instability is not a cause of variable bioavailability. PMID- 15544058 TI - Physical and chemical stability of different formulations with superoxide dismutase. AB - Topical formulations with superoxide dismutase (SOD), a scavenger of superoxide radicals, have proved to be effective against some skin diseases. Nevertheless, formulations with proteins are susceptible to both chemical and physical instability. Three different formulations (anionic and non-ionic gel and emulsion) were developed and supplemented with SOD in order to determine the most stable formulation that would maintain SOD activity. Physical stability was evaluated by assessing the rheological behavior of the formulations stored at room temperature, 37 and 45 degrees C. Chemical stability was evaluated by the measurement of enzymatic activity in the formulations stored at room temperature and at 45 degrees C. Formulations showed a flow index less than one, characterizing pseudoplastic behavior. There was no significant difference in initial values of flow index, tixotropy or minimum apparent viscosity. Neither gel showed significant changes in minimum apparent viscosity concerning storage time or temperature, as well, SOD presence and its activity. The emulsion showed decreased viscosity by the 28th day, but no significant changes concerning storage temperature or SOD presence, although it showed a decreased activity. The addition of SOD to the formulations studied did not affect their physical stability but gel formulations seem to be better bases for enzyme addition. PMID- 15544059 TI - Sustained-release hydroxycamptothecin polybutylcyanoacrylate nanoparticles as a liver targeting drug delivery system. AB - Sustained release hydroxycamptothecin polybutylcyanoacrylate nanoparticles (HCPT PBCA-NP) associated with polybutylpyrolidone (PVP) for liver targeting were prepared by the adsorption-entrapping method. The morphology, sizes, drug loading efficiencies, release characteristics in vitro, distribution and pharmacokinetic parameters in vivo of the HCPT-PBCA-NP associated with PVP were studied. The median diameter of the particles was 81 nm and the drug loading was 1.2%. The release characteristics in vitro were in accordance with the Higuchi equation: Q = 0.0615 + 0.0940 mean square root t. 64.5% of the HCPT were concentrated in the liver at 15 min after i.v. administration of HCPT-PBCA-NP associated with PVP. The plasma drug concentration-time curve of the HCPT in rabbits was fitted to a two-compartment open model. The Vc, T 1/2 and CL were 3.5L; 147h; 0.18L x h(-1), respectively. The method of preparation presented in this paper seems to bean alternative for the preparation of PBCA-NP of poorly soluble drugs both in water and in lipid. PMID- 15544060 TI - A comparison of the efficacy of a bispyridinium oxime--1,4-bis-(2 hydroxyiminomethylpyridinium) butane dibromide and currently used oximes to reactivate sarin, tabun or cyclosarin-inhibited acetylcholinesterase by in vitro methods. AB - The efficacy of a bispyridinium oxime 1,4-bis(2-hydroxyiminomethylpyridinium) butane dibromide, called K033, and of currently used oximes (pralidoxime, obidoxime, oxime HI-6), to reactivate acetylcholinesterase inhibited by various nerve agents (sarin, tabun cyclosarin) was tested by in vitro methods. The new oxime K033 was found to be a more efficacious reactivator of sarin or cyclosarin inhibited acetylcholinesterase than pralidoxime and obidoxime but it did not reach the efficacy of oxime HI-6 in the case of the inhibition of acetylcholinesterase by sarin or cyclosarin. On the other hand, oxime K033 was more efficacious than oxime HI-6 in reactivating tabun-inhibited acetylcholinesterase. Thus, oxime K033 seems to be a relatively efficacious broad spectrum acetylcholinesterase reactivator and, therefore, could be useful if no information about the type of nerve agent used was available. PMID- 15544061 TI - In vitro alpha amylase inhibitory effect of some clinically-used drugs. AB - Twenty six medicinal compounds used for treating various diseases and representing different chemical and pharmacological classes were tested for their potential alpha amylase inhibitory activity in vitro. Three of these (tetracycline, enalapril and captopril) were found to have a substantial alpha amylase inhibitory activity. The effect was shown to be dose dependent and the IC50 was determined (tetracycline = 0.59 mM, enalapril = 0.29 mM, and captopril = 0.78 mM) and compared to that of acarbose (0.0062 = mM). The potential pharmacological implications of this effect are discussed. PMID- 15544062 TI - In vitro effects on proliferation, telomerase activity and apoptosis of an eremophilanoid sesquiterpene from Senecio oldhamianus maxim in cultured human tumor cell lines. AB - 8,11-Dioxol-6-en-9alpha, 10alpha-epoxy-8beta-hydroxyeremophilane (HEM), a new eremophilanoid sesquiterpene, was isolated from Senecio oldhamianus Maxim. Its effects of cytotoxicity, telomerase activity, apoptosis and related genes expression in two human tumor cell lines, human hepatoma cells SMMC-7721 and human oophoroma cells HO-8910, were studied. Hydroxycamptothecine (HCPT) was used as a positive control. The IC50 of cytotoxicity by HEM were 24.9 +/- 2.1 and 19.4 1.6 microM in SMMC-7721 and HO-8910 cells respectively, and 0.35 +/- 0.10 and 0.27 +/- 0.08 microM for HCPT. HEM inhibited telomerase activity with the IC50 35.9 +/- 3.2 microM in SMMC-7721 and 25.6 +/- 2.6 microM in HO-8910 cells, while HCPT had no effect on telomerase activity in both tumor cell lines. HEM 20-30 microM induced apoptosis in SMMC-7721 cells from 5.7% to 18.4% and in HO-8910 cells from 7.6% to 67.1%. While HCPT 0.1-0.5 microM induced apoptosis in SMMC 7721 cells from 6.5% to 13.3% and in HO-8910 cells from 9.9% to 30.9%. HEM 30 microM significantly decreased Bcl-2 protein expression to 58.7% in SMMC-7721 and to 57.6% in HO-8910 cells. While HCPT 0.5 microM significantly decreased Bcl-2 protein expression to 64.3% in SMMC-7721 and to 70.0% in HO-8910 cells. HEM 25 microM and 30 microM significantly increased P53 protein expression 2.3-3.6-fold in SMMC-7721 and 3.0-5.7- fold in HO-8910 cells. While HCPT 0.5 microM significantly increased P53 protein expression 3.3-fold in SMMC-7721 and 2.7-fold in HO-8910 cells. Overall, HCPT exhibited a more potent effect on cytotoxicity and apoptosis in the two tumor cell lines than HEM did. However HEM can inhibit telomerase activity in the two tumor cell lines but HCPT cannot. PMID- 15544063 TI - A new furobenzopyranone and other constituents from Anaphalis lactea. AB - Together with twenty-one known compounds, a new furobenzopyranone was isolated from the whole plant of Anaphalis lactea. Their structures were elucidated by spectroscopic methods MS, IR, UV, NMR, including 2D-NMR techniques. The anti bacterial activity of compounds 1, 4-6, 14, 15 and the anti-tumor activity of compounds 4-6 were tested. PMID- 15544064 TI - Liposomal amphotericin B dry powder inhaler: effect of fines on in vitro performance. AB - The aim of the present investigation was to improve in vitro pulmonary deposition of amphotericin B (AMB) liposomal dry powder inhaler (LDPI) formulations. Liposomes with negative (AMB1) and positive (AMB2) charge were prepared by the reverse phase evaporation (REV) technique, extruded to reduce size, separated from unentrapped drug and lyophilized using an optimized cryoprotectant to achieve maximum drug retention. Lactose carrier (Sorbolac 400) in varying mass ratio with or without addition of fines (500# sieved Pharmatose 325M) in different mixing sequence were used to formulate AMB LDPI formulations. In vitro evaluation was done with twin stage impinger (TSI) for fine particle fraction. The lactose carrier containing 10% fines was found to be optimum blend at 1:6 mass ratio of liposome: lactose. The addition of fines and order of mixing fines were found to influence the fine particle fraction (FPF) significantly. FPF of LDPI formulations using a Rotahaler (Cipla, India) as delivery device at 30, 60 and 90 L/min were found to be 23.1 +/- 1.5 percent and 17.3 +/- 2.2 percent; 25.3 +/- 1.8 percent and 19.6 +/- 1.5 percent and 28.4 +/- 2.1 percent and 22.9 +/- 1.9 percent for AMB1 and AMB2 respectively. PMID- 15544065 TI - Dissolution test for ivermectin in oral veterinary paste. AB - A dissolution test for oral veterinary pastes with ivermectin using the Ph. Eur. paddle apparatus was developed. Sink conditions were achieved with sodium lauryl sulphate in a concentration of 0.5% as dissolution medium. By means of HPLC fast degradation of ivermectin was observed in HCl 0.1 M solution. Rotation speed of the paddle at 75 rpm was appropriate as demonstrated in a study comparing two different products. PMID- 15544067 TI - BMC improves patient flow throughout the hospital. PMID- 15544066 TI - Chemical properties of a mannoglucan from Cistanche deserticola. AB - A new mannoglucan from Cistanche deserticola from China is characterized. The compound is responsible for a mild stimulation of mitogen-induced T and B lymphocyte proliferation. PMID- 15544068 TI - New OR streamlines surgeries, increases caseload. AB - The number of surgeries using minimally invasive equipment has been growing rapidly at Children's Hospital of Pittsburgh, as both referring physicians and parents seek out surgeons using techniques that allow for quicker recovery and less scarring. In 2000, there were 276 minimally invasive surgeries performed at CHP. By 2003, the number had grown to 514 and so far in 2004 almost 700 such cases have been done. PMID- 15544069 TI - Implement balanced scorecard to translate strategic plan into actionable objectives. AB - Faced with challenges ranging from declining reimbursement to staff shortages, health care organizations--integrated delivery systems, physician group practices, disease management providers, and others--increasingly are turning to general business models to map out step-by-step action plans for performance measurement and process improvement. Creating a "balanced scorecard" is an obvious starting point for assessing and improving clinical and financial performance. PMID- 15544070 TI - Four factors that lead to performance improvement in hospitals. AB - What sets high-performing hospitals apart from others? Is it a great quality improvement department or superior executive leadership? Is it hiring employees who are dedicated to finding better ways to doing their jobs? A new report from The Commonwealth Fund says there is no one thing that makes a hospital a performance leader. Rather, the report identifies four key elements that combine to make certain hospitals stand out from the rest of the pack. PMID- 15544071 TI - Diagnostic approach to the patient with jaundice following trauma. AB - BACKGROUND AND AIMS: Jaundice in trauma patients may reflect serious underlying pathology. The aim of this review was to determine the appropriate diagnostic approach to the patient with jaundice following trauma. METHODS: A MEDLINE search was performed to retrieve publications which outlined the causes of jaundice in trauma patients. RESULTS: The main causes of jaundice in trauma patients were found to be bilirubin overload caused by breakdown of transfused- and extravasated blood and hepatic dysfunction caused by sepsis, infections, initial shock and systemic hypotension. Bile duct injury or drug induced liver injury are rare. Liver function tests are often uninformative but commonly show a cholestatic pattern. Ultrasound, CT or ERCP are the diagnostic imaging methods most widely used. Abdominal ultrasound and CT may reveal specific organ injuries, bile duct dilatation, intraabdominal fluid collections, hematomas or acalculus cholecystitis. ERCP is often diagnostic and permits a therapeutic intervention when a bile duct injury is present. CONCLUSIONS: The primary aim of the diagnostic approach should be to identify all cases of bile duct injury or obstruction. Sepsis and infections should be actively looked for. The number of blood transfusions must be calculated. Ultrasound, CT or ERCP are the diagnostic imaging methods most widely used. PMID- 15544072 TI - The role of biofeedback therapy in functional proctologic disorders. AB - BACKGROUND AND AIMS: The question which patients with functional proctologic disorders truly benefit from the biofeedback has not been equivocally resolved. The aim of this study was to assess our results of biofeedback therapy in patients with anal incontinence or constipation. MATERIAL AND METHODS: Fifty-two consecutive patients who were treated with biofeedback therapy between January 1998 and March 2002 were studied. Data was collected from our proctologic database. RESULTS: Of the twenty-two patients with anal incontinence who underwent biofeedback therapy during the study period, twenty patients had incontinence affecting quality of life. Twelve patients (60 percent) benefited from biofeedback as judged by improvement of incontinence symptoms affecting quality of life; all four patients with partial sphincter defects, three out of four patients after secondary repair, three out of five patients with persistent incontinence after rectal prolapse surgery and two out of seven patients having idiopathic incontinence. Of the thirty patients who underwent biofeedback therapy for constipation, twenty-five had intractable symptoms of constipation. Constipation resolved in sixteen patients (64 percent); in thirteen out of nineteen (68 percent) of those with pelvic floor dysfunction (PFD) and in three out of six (50 percent) having combined PFD and slow transit constipation. In patients with PFD constipation was resolved in ten out of thirteen patients (77 percent) with anismus but in only three out of six (50 percent) having other causes. CONCLUSIONS: Biofeedback therapy improves incontinence after sphincter repairs and in patients with partial external sphincter defects, but does not improve idiopathic incontinence. Biofeedback is also effective in patients with constipation, especially when anismus is the only cause for symptoms of constipation and difficult evacuation. PMID- 15544073 TI - Is positron emission tomography using 18F-fluorodeoxyglucose and 11C-acetate valuable in diagnosing indeterminate pancreatic masses? AB - BACKGROUND: It can be impossible to differentiate a mass forming chronic pancreatitis from adenocarcinoma of the pancreas using standard anatomical imaging. Positron emission tomography using 2-[18F] fluoro-2deoxy-D-glucose (18FDG-PET) and 1-[11C]-acetate (11C-acetate-PET) are methods taking advantage of the metabolic differences between benign and malignant tissues. AIMS: To determine the diagnostic accuracy of 18FDG-PET and 11C-acetate-PET in indeterminate pancreatic masses. METHODS: Twenty patients with an indeterminate mass of the head of the pancreas were prospectively studied. All patients underwent 18FDG-PET and eighteen of them 11C-acetate-PET. Scans were evaluated qualitatively and quantitatively; the later by using regional standardised uptake value (SUV). Final diagnosis was established using histopathologic evaluation of resected specimen or biopsy. RESULTS: Adenocarcinoma was diagnosed in twelve patients and chronic pancreatitis in eight. Qualitative evaluation of 18FDG-PET imaging revealed three false negative and one false-positive results. The sensitivity, specificity, and diagnostic accuracy were 75 %, 88 %, and 80 %, respectively. The cut-off SUV to differentiate malignant from benign disease was 3,5 demonstrating a sensitivity of 91.7 % and a specificity of 75 %. CONCLUSION: 18FDG-PET imaging could not confirm or exclude malignancy in indeterminate masses of the head of the pancreas with high sensitivity and diagnostic accuracy. 11C acetate-PET provided no additional diagnostic benefits. PMID- 15544074 TI - Epidemiology of severely and fatally injured patients in western part of Norway. AB - BACKGROUND AND AIMS: Analysis of the injury mechanism and characteristics of severely and fatally injured patients in the western part of Norway. MATERIAL AND METHODS: We did a prospective registration of all severely injured patients hospitalized during a three-year period. The files of severely injured patients that died at scene or during transport were retrospectively retrieved from the Forensic Department. A total of 558 patients with an Injury Severity Score > 15 were included. RESULTS: Four-hundred-forty-four men (79.6 %) and 114 women (20.4 %) with a median age of 36 and 51.5 years respectively were included. The proportion of female patients older than 80 years were 19.3 % compared to 5.6 % for men. A total of 215 (38.5 %) patients died and 149 (69.3 %) of these patients died on scene or during transport. The incidence of severely injured patients in Hordaland County was 30 per 100,000 inhabitants per year. The incidence was lowest among children below 5 years (7/100,000/year) and highest among persons older than 80 years (95/100,000/year). Men had a 3.8 times greater risk of getting seriously injured compared to women. Road traffic accidents were the cause of the injuries in 235 (42.1 %) patients and 35.8 % of these patients died. A total of 215 (38.5 %) patients were injured due to falls and 30.2 % of these patients died. Patients who had sustained falls were significantly older than patents with other injury mechanisms (p < 0.001, CI = 13.0-20.2). The proportion of patients with penetrating injuries was 7.3 %. CONCLUSION: The incidence of severely injured patients was 30/100000/year. Men and elderly people had a higher risk of getting severely injured. Falls were the dominating injury mechanism among elderly. PMID- 15544075 TI - Severe secondary peritonitis following gastrointestinal tract perforation. AB - BACKGROUND AND AIMS: Early prognostic evaluation of abdominal sepsis is useful in the assessment of the severity of the disease and to select high-risk patients for early surgical reintervention. The aim of this study was to identify prognostic factors in a well-defined patient population most likely to benefit from early reoperation. MATERIAL AND METHODS: Retrospective analysis of 66 consecutive patients with secondary peritonitis caused by gastrointestinal tract perforation and requiring postoperative treatment in an intensive care unit was performed using univariate and multivariate analysis to identify risk factors for hospital mortality. RESULTS: The overall hospital mortality rate was 36 %. Significant risk factors in the univariate analysis included advanced age (p = 0.000), pre-existing illness (p = 0.000), chronic medication (p = 0.028), hospital transfer (p = 0.036), non-traumatic cause of perforation (p = 0.031), high Mannheim peritonitis index (MPI) score (p = 0.001), and high C-reactive protein (CRP) level in the early postoperative phase (p = 0.015). In a multivariate analysis, only advanced age (odds ratio 1.1008, p = 0.000) and high postoperative CRP level (odds ratio 1.0095, p = 0.008) were identified as independent prognostic factors for hospital mortality. CONCLUSION: In addition to factors associated with the physiological reserve of the patient, type of peritonitis and high MPI score, elevated CRP levels in the early postoperative phase in patients operated for severe secondary peritonitis have prognostic significance. However, before a properly designed randomized study on the value of planned relaparotomy in secondary peritonitis can be initiated, more reliable methods to identify high-risk patients need to be found. PMID- 15544076 TI - The effect of fibrin sealant combined with fibrinolysis inhibitor on reducing the amount of lymphatic leakage after axillary evacuation in breast cancer. A prospective randomized clinical trial. AB - BACKGROUND AND AIMS: One third of women undergoing mastectomy with axillary evacuation for primary breast cancer suffer from postoperative seromas leading to unnecessary costs and complications such as infections and new operations. Different methods to prevent seroma formation have been tried without permanent success. The aim of this prospective randomised study was to examine the effect of fibrin sealant with fibrinolysis inhibitor firstly on the reduction of the amount of lymphatic leakage after axillary evacuation and secondly on the reduction of days with drains and postoperative seroma punctures. METHODS: 40 patients with primary breast cancer were prospectively randomised to the treatment group (n = 19) getting fibrin glue combined with fibrinolysis inhibitor (aprotinin) sprayed into the axillary fossa and to the control group (n = 21). RESULTS: There were no differences in the incidence of postoperative seromas between the groups. However, the seromas were easier to treat if fibrin clue was used. Total quantity (mean+/-SD) of lymphorrhea and total number of aspirations (mean+/-SD) were almost twice as high in the patients of the control group compared to those having fibrin sealant. In the treatment group seromas resolved after one or occasionally after two aspirations in 71 % of patients, while in the control group 90 % of patients needed three or more aspirations. CONCLUSION: Potentially, fibrin sealant combined with fibrinolysis inhibitor might be used for the treatment of post- axillary evacuation lymphorrhea and seroma. PMID- 15544077 TI - Cavoatrial extension of renal cell cancer: results of operative treatment in Helsinki University Hospital between 1990 and 2000. AB - BACKGROUND AND AIMS: To evaluate the technical procedures and the post-operative survival of patients having been operated for renal cell cancer with cavoatrial tumour thrombus (RCC-T). MATERIAL AND METHODS: Between 1990 and 2000 the cardiac unit at Helsinki University Central Hospital operated on seven patients for RCC T. A cardiac surgeon along with a urologist, performed all seven operations using sternolaparotomy (either midline or Chevron incision) with cardiopulmonary bypass. RESULTS: The average duration of the operations was eight hours (range 6 11 hours) and the average perfusion time was 118 minutes (range 35-206). Hypothermic circulatory arrest was used for one patient with an arrest time of 31 minutes. Only with one patient could the cavotomy be closed directly. In four patients a cava resection was performed and closed either with a pericardium patch or a Gore-Tex prosthesis. In two patients the cava was ligated below the renal veins. During the post-operative intensive care, there were two deaths. Of the remaining patients, five were alive after six months, four after 12 months, three after six years and one patient is still alive after 12 years of follow-up. CONCLUSIONS: In agreement with previously published results, although peri operative mortality is relatively high with RCC-T patients, long-term post operative survival is possible. PMID- 15544078 TI - Does technical failure of revascularization during coronary artery bypass grafting predict severity of poststernotomy mediastinitis? AB - BACKGROUND AND AIMS: To assess the impact of unsuccessful revascularization in relation to poststernotomy mediastinitis (PSM), which affects long-term outcome after coronary artery bypass grafting (CABG). MATERIAL AND METHODS: An active approach for the follow-up of PSM involved a step by step treatment protocol of conventional surgery and plastic reconstructive surgery. 47 patients treated for PSM after CABG were identified and further evaluated. Complete revascularization was considered unsuccessful when technical hazards were reported during CABG. When PSM subsided after thorough debridement and sternal refixation without plastic reconstructive surgery, such as omentoplasty or muscle transposition, PSM was categorized as mild PSM. If treatment required plastic reconstructive surgery, PSM was categorized as severe PSM. Preoperative coronary artery angiographic status and success of revascularization were compared to postoperative outcome in relation to mild and severe PSM. RESULTS: 36 patients suffered from mild PSM and 11 patients from severe PSM. Preoperative clinical status did not differ among patients. Two patients (4.3 %) died during hospitalization. The need for plastic reconstructive surgery was significant (p < 0.05) among patients with unsuccessful revascularization. 35 out of 41 patients (85 %) without problems of graft anastomosis during CABG (successful revascularization) were associated with mild PSM, whereas only 6 out of 41 patients (15 %) with successful revascularization during CABG required plastic reconstructive surgery (p < 0.05). Technical failure of graft anastomosis (3 cases) or poor outflow of internal thoracic artery (2 cases) were statistically associated with severe PSM. CONCLUSION: Technical failures of revascularization during CABG may delay recovery from PSM. PMID- 15544080 TI - Primary cartilage lesions of the knee joint in young male adults. Overweight as a predisposing factor. An arthroscopic study. AB - BACKGROUND AND AIMS: This study was undertaken to evaluate the relationship between overweight and severity of arthroscopically confirmed primary cartilage lesions of the knee. MATERIAL AND METHODS: The hospital records of 88 young male adults (98 knees), who underwent arthroscopy of the knee and were diagnosed of primary cartilage lesion during 1997-98, were reviewed. The depth of the lesions was graded according to Beguin and Locker classification. RESULTS: 73.5 % of the lesions were patellar and 12.0 % in the medial condyle of the femur. 74.5 % of the patients had superficial (grade I-II) and 25.5 % deep (grade III-IV) lesions. Patients with deep lesions had a significantly higher body mass index (BMI 25.3 vs. 22.9, p < 0.001) and they were older (20.8 vs. 19.7 years, p = 0.023) than other patients. Similarly, patients with overweight (BMI > or = 25.0) had significantly more often deep lesions than other patients (50 % vs. 10.4 %, p < 0.001). CONCLUSIONS: These results suggest, that overweight may predispose young patients to more severe cartilage lesions independent of other etiologic factors, and support the hypothesis of the cumulative effect of overweight on cartilage injuries during early adult life. PMID- 15544079 TI - Unstable trochanteric fractures augmented with calcium phosphate cement. A prospective randomized study using radiostereometry to measure fracture stability. AB - BACKGROUND AND AIMS: Internally fixed unstable trochanteric fractures might be difficult to retain in position during healing. Secondary displacement might lead to malunion and poor functional result. The aim with this study was to measure whether augmentation with resorbable calcium-phosphate cement could improve fracture stability as shown in biomechanical studies. MATERIAL AND METHODS: 26 ambulatory patients with an unstable trochanteric fracture were randomized to treatment with a sliding screw device alone (Controls) or the same device combined with calcium-phosphate cement for augmentation (Augmented). All patients were allowed unrestricted weight bearing after surgery. Fracture movement was measured with radiostereometry (RSA) at 1 and 6 weeks and at 6 months. RESULTS: Two patients died during the study period due to unrelated causes and another three were excluded due to technical problems with the RSA in two and concomitant illness in one. 21 patients (11 Augmented and 10 Controls) were followed according to the study protocol. At 1 week the augmented fractures had moved on average 1.9+/-1.7 mm while movement in the controls was 4.0+/-2.4 mm (p < 0.05). The average total movement from the day after surgery until 6 months, when all fractures had healed, was 7.8+/-6.2 mm for the augmented fractures and 13.2+/-4.3 mm for the controls (p < 0.05). Varus angulation was the most pronounced rotational movement for both groups although augmented fractures revealed less varus angulation compared with controls at all time points. Rotation around the longitudinal and transversal axes were small with no significant differences between treatment groups. CONCLUSION: Augmentation with calcium-phosphate cement improved the stability of unstable trochanteric fractures fixed with a sliding screw device. The improvement was most pronounced for varus angulation and lateral and distal migration of the head and neck fragment. PMID- 15544081 TI - Hip fractures in Finland--a comparison of patient characteristics and outcomes in six hospitals. AB - BACKGROUND AND AIMS: To compare six Finnish hospitals for the quality of treatment of hip fractures and to obtain information for the development of care. MATERIAL AND METHODS: Data of 1179 consecutive hip fracture patients (about 200 patients per hospital) was collected prospectively, using similar standardized forms and focusing on background factors and the four-month functional outcome. RESULTS: There were significant differences between the hospitals in patient characteristics (age, place of residence, walking ability, use of walking aids, morbidity and type of fracture) and in the unadjusted outcome variables at four months' follow-up (place of residence, mobility, use of walking aids and pain in injured hip). After adjustment for baseline characteristics, there was a significant difference in the post-fracture walking ability between the centres but no significant differences in post-fracture place of residence. Unadjusted mortality did not vary between the centres, but adjustment resulted in significant differences. The most marked difference in surgical methods between the hospitals was seen in the use of either sliding hip screw or Gamma Nail for trochanteric fractures, but this difference was not reflected in the results of multivariate analysis. CONCLUSIONS: We found minor differences in mobility and mortality between the participating hospitals, and these might serve them as a stimulus for improving their standard of good practice. Continuous quality improvement by repeating the audit cycle is recommended in order to reach and then improve the prevalent standards in the care of hip fracture patients. Confounding factors should be adjusted when comparing the medical centres treating hip fractures, and the evaluation of the results should be multidimensional. PMID- 15544082 TI - [Disappearance of differences in antibiotic resistance of Staphylococcus aureus strains isolated in hospitals and in general practice]. AB - There is general opinion that Staphylococcus aureus strains isolated in hospitals are more frequently resistant to antibiotics than community strains, however, the increasing resemblance between hospital and community strains has been recently reported. The aim of the study was to compare the antibiotic resistance and phage type pattern of S. aureus strains isolated from patients treated either in hospitals or in general practice in northern part of Poland. The study was conducted on 771 S. aureus strains isolated from different specimens. Phage typing was performed according to the method of Blair and Williams. The drug susceptibility was determined by the disc-diffusion method. There were no significant differences in antibiotic resistance or phage-type pattern when hospital and community methicillin-sensitive S. aureus (MSSA) strains were compared. The most MSSA were resistant to penicillin (84.6% and 82.1% respectively) and doxycycline (49.3% and 50.4% respectively) whereas they were rarely resistant to other antibiotics. The predominance of phage group II was found in both hospitals (28.0%) and general practice (29.9%). Phage group III, usually associated with hospitals, occurred in small percentage (12.9% and 9.4% respectively) while to this group predominantly (76.6%) multiresistant methicillin resistant S. aureus (MRSA) isolated in hospitals belonged. These results suggest, that there is only slight difference in antibiotic resistance between hospital and community S. aureus strains. Antibiotic resistance pattern mainly results from frequency of appearance of MRSA, mostly occurring in hospitals. PMID- 15544083 TI - [Determination of methicillin resistance in clinical strains of Staphylococcus aureus in routine microbiological diagnostics]. AB - The aim of this study was to compare the results of two methods: disc-diffusion test and ATB STAPH 5 (version 2000) test applied for determination of methicillin resistant staphylococci. One hundred and twenty S. aureus strains isolated from clinical specimens in microbiological laboratory of Medical University in Warsaw were investigated. Identification and drug resistance were determined using automatic ATB Expression System. We compared the results of methicillin-resistant S. aureus detection by disc-diffusion method (oxacillin 1 microg) and ATB STAPH 5; version 2000 (bioMerieux sa). In the case of 116 strains S. aureus identical results were obtained in both methods, 4 strains gave inconsistent results in these two methods. Results that were obtained show that the ATB STAPH 5 (version 2000) test is as useful for routine determination of MRSA and MSSA strains as the disc-diffusion method, and may be used alternatively. PMID- 15544084 TI - [The use of Multiplex PCR-RFLP method (multilocus RFLP) in identification and differentiation of enterococci]. AB - The present study proposed use of modified multilocus sequence typing (MLST) approach, for subspecies differentiation of Enterococcus faecalis isolates. We disgesed with SspI endonuclease product of amplification of three E. faecalis antigen-encoding genes (ace, encoding a collagen and laminin adhesin; efaA, encoding an endocarditis antigen; and salA, encoding a cell wall associated antigen) and one housekeeping gene (pyrC) of 32 E. faecalis isolates MLRA analysis of polymorphic regions of these four genes identified 23 distinct types 21.4 to 100% identity among the 32 isolates. Further studies are needed to evaluate the value of proposed method. PMID- 15544085 TI - [Occurrence of Corynebacterium amycolatum strains in clinical specimens]. AB - C. amycolatum is poorly recognized and rarely described in the world literature. So, better recognizing and understanding biology of these bacteria may help effectively prevent infections caused by them. The subject within the study were 70 of C. amycolatum strains which were isolated from the clinical specimens of patients hospitalized at the State Clinical Hospital in Bydgoszcz. After initial identification of examined strains based on Gram staining results, colonial morphology, biochemical and enzymatic features included in API Coryne and API ZYM tests (bioMerieux), growth at 20 degrees C, Tween 80 requirement, DNA and tyrosine hydrolysis, occurrence in clinical specimens and origin of C. amycolatum strains were analyzed. The investigated strains were the most frequently isolated from wound swabs (61.5%), urine (14.3%), drain swabs (7.1%) and mainly (37.2%) came from patients treated at the departments of surgery. PMID- 15544086 TI - [Biochemical properties of Corynebacterium amycolatum strains]. AB - C. amycolatum is the most commonly isolated nonlipophilic species of Corynebacterium from clinical samples. However, the lack of good commercial identification tests in microbiology laboratories causes some difficulties in C. amycolatum diagnostics. We decided to examine biochemical and enzymatic properties of isolated strains and analyze the occurrence of particular biochemical profiles (biotypes). Perhaps it would let improve the identification schemes. 70 strains of C. amycolatum were analyzed. The estimation of biochemical properties consisted of the results of API Coryne and API ZYM tests (bioMerieux), the ability of excreting of protease, esterase, lipase and lecithinase. Analyzed strains had various biochemical and enzymatic properties. Almost all strains fermented glucose (98.6%) and maltose (95.7%) and produced pyrasinamidase (94.3%). All strains produced alkaline phosphatase and phosphohydrolase, and 95.7%--acid phosphatase. Biotypes of particular strains were determined on the biochemical reactions included in the API Coryne tests. In the group of 70 strains 21 profiles were distinguished among which 3100325 biotype (35.7%) was dominant. The lipolysis was defined on Tween 20, Tween 40, Tween 60, Tween 80 medium and with the API ZYM test usage. All strains produced esterase-lipase (esterase C-8), 95.7% of strains-esterase C-4, and 21.4% lipase C-14. Among analyzed strains 18.6% hydrolyzed Tween 20, 14.3% Tween 60, and 1.4% Tween 40. None of these strains demonstrated lipase and lecithinase activity. Difficulties in concerning C. amycolatum as pathogens justify further investigations. PMID- 15544087 TI - [Comparative study of thermoresistance spores of Clostridium difficile strains belonging to different toxigenicity groups]. AB - The thermoresistance of spores of Clostridium difficile strains belonging to the different toxigenicity groups was compared in the study. Among spores of toxigenicity C. difficile strains (26 C. difficile strains produced toxins A and B (TcdA+TcdB+) and 32 C. difficile strains produced only toxin B (TcdA-TcdB+) were high thermoresistant. Between spores of non-toxigenic C. difficile strains much lower thermoresistance was observed. In conclusion, more studies are needed to clarify the importance of spores transmission in the increasing number of AAD cases in Poland. PMID- 15544088 TI - [Detection of the expression of extended-spectrum beta-lactamases (ESBL) in clinical strains of Klebsiella pneumoniae and Escherichia coli: comparison of E test and two-disc methods]. AB - The aim of this study was to compare the effectiveness of E-test and two disc methods applied for the detection of extended spectrum beta-lactamases. All strains were tested by E-test, by double-disc synergy test (DDST) according to Jarlier (cefotaxim, ceftazidim, aztreonam and clavulanic acid) and also by disc test according to Appleton (cefpodoxime and cefpodoxime with clavulanic acid, CPD and CD01 disc). We tested 148 clinical strains of E. coli and 78 strains of K. Pneumoniae. In case of K. pneumoniae, the activity of the ESBLs was detected among 30 strains--both in E-test, Jarlier test and Appleton test. Among E. coli, four strains were found ESBL-positive in the test according to Jarlier but only three strain of these when E-test and Appleton test was used. The results of investigations performed suggest, that E-test and disc methods according both Jarlier and Appleton have the same effectiveness in detection ESBLs among K. pneumoniae strains. However, in case of E. coli, interpretation of results may present a problem. PMID- 15544089 TI - [Using the PCR-RFLP method for comparative analysis of the pathogenic Escherichia coli strains]. AB - The purpose of the study was to identify differences and similarities between Escherichia coli strains which do or do not utilize disaccharide sucrose by PCR RFLP method. Investigations were done on chromosomal DNA level using cscA gene associated with conservative sequences. The cscA gene may be found in all of the analysed strains. Genotypic analysis demonstrated presence of the same restriction model consisted of 2 DNA fragments with size of 161 bp and 110 bp in all of E. coli strains. Results of these investigations have shown that there are no differences between E. coli strains. PMID- 15544090 TI - [Adhesion of Acinetobacter spp. to para-xylene]. AB - Hydrophobicity of 200 of Acinetobacter spp. strains was defined by the BATH (Bacterial Adhesion to Hydrocarbon Test) method of Rosenberg et al. 48.0% strains of Acinetobacter spp. adhered to para-xylene. 40.0% of strains showed waek hydrophobicity, 7.5% moderate and one strain strongly adhered to para-xylene. A. baumannii strains adhered in 52.2%, A. junii in 42.1% and 20.0% in A. haemolyticus. Significantly we find weak adhesion to para-xylene than mild and strong. Mostly adhesion was discovered in A. baumannii and A. junii than in A. haemolyticus. Results show that Acinetobacter spp. has adhesive properties to para-xylene. PMID- 15544091 TI - [Interaction of neomycin with other antibiotics on selected bacterial strains]. AB - Antimicrobial combinations are used most frequently to provide broad-spectrum empirical coverage in the treatment of bacterial infections. However, combination of two antibiotics may not influence their activity, may lead to synergy or antagonism in the activity. Neomycin may be combined with one of the following antibiotics: ampicillin, procaine penicillin, gramicidin, bacitracin, polymyxin B, lincomycin, oxytetracycline, and erythromycin in some human and veterinary multiantibiotic drugs distributed in Poland. The checkerboard method has been one of the traditional assays for the measurement of antibiotic interactions. The aim of this study was to analyse the activity interaction of neomycin with second antibiotic in multiantibiotic drugs distributed in Poland on standards and clinical bacterial strains. Checkerboard results for all strains demonstrated synergism for 2.5% of combinations, only for standards strains. In one case Salmonella Enteritidis, in combination of neomycin with bacitracin, inhibition effect was observed. Additive effects were predominant--49%. In 18% neutral effects were shown, but in 26% of combinations FIC indexes were not possible to calculate, because of the resistance of clinical strains to the highest concentration of at least one antibiotic. In combination of aminoglycoside (neomycin) with beta-lactams antibiotics (ampicillin, procaine penicillin) in vitro, no synergy was observed for all examined strains. The best results were achieved for combinations of neomycin with peptide antibiotics (polymyxin, gramicidin and bacitracin)--5 for all 6 synergy effect observed. PMID- 15544092 TI - [Evaluation of selected alcoholic antiseptics activity against clinical bacterial strains]. AB - The aim of this study was to evaluate antimicrobial activity of selected alcoholic antiseptics against clinical strains, which possessed in majority a high level of drug resistance: MRSA (7), MSSA (3), E. coli: (9): strains producing ESBL (4), P. aeruginosa: (4), E. cloacae: (3), K. pneumoniae: (3). These strains were defined by MIC value, using antibiotic agar dilution method according to NCCLS. Fourteen alcoholic antiseptics were used in this study. Beside alcohol, they contained other active substances like iodine, hydrogen peroxide, chlorhexidine. Some additional agents were included for easier application, such as: gelling, moisturizing, aromatic or coloring substances. The objective of this study was also to determine the dependence of bactericidal activity on preparations (concentration). Product undiluted and diluted two and four times in water was analyzed according to prEN 12054 standard; 30 seconds and 1 minute contact time was used. The obtained data indicate that all tested undiluted antiseptics possessed bactericidal activity described by producers. However antiseptics (dilution leads to decrease and even loss of bactericidal activity. Two-times dilution of gel almost completely inactivated the product. Antimicrobial activity after 30 seconds of contact time was not affected by presence of additional agents in the tested antiseptics. PMID- 15544093 TI - ["We don't want anything to do with an undervalued school!" Interview by Nathalie Debertrand]. PMID- 15544094 TI - [Bioethics law and advances in reproductive technology]. PMID- 15544095 TI - [What management for premature infants?]. PMID- 15544096 TI - [From burn to cure. Multidisciplinary management]. PMID- 15544097 TI - [Prevention and emergency care of burns in children]. PMID- 15544098 TI - [Management of pain in burned children]. PMID- 15544099 TI - [Care of children in a burn unit]. PMID- 15544100 TI - [Play therapy to respond to the needs of the burned child]. PMID- 15544101 TI - [Psychological management of the burned child and his family]. PMID- 15544102 TI - [Management of the burned child in the rehabilitation center]. PMID- 15544103 TI - [The pediatric nurse in Burkina Faso]. PMID- 15544104 TI - [1/6 Administration of a sugar solution by sucking--maternal lactation]. PMID- 15544105 TI - [4/5 Nurses' aides and patient education]. PMID- 15544106 TI - [Procalcitonin--acute inflammatory marker in infections]. PMID- 15544107 TI - [Dual blockade of the renin-angiotensin system--is it of clinical significance?]. PMID- 15544108 TI - [Dual blockade of the renin-angiotensin system]. PMID- 15544109 TI - [Procalcitonin as a marker of severe bacterial infection in children]. PMID- 15544110 TI - [Autosomal dominant polycystic kidney disease. Proposal to guidelines for patients and their relatives]. PMID- 15544111 TI - [Monogenetic obesity with special focus on the melanocortin 4 receptor gene mutations]. PMID- 15544112 TI - [Pharmacotherapy of obesity]. PMID- 15544113 TI - [Diagnosis of ischemic heart disease. Exercise ECG versus myocardial scintigraphy and coronary angiography]. PMID- 15544114 TI - [Paracetamol poisoning in teenagers]. PMID- 15544115 TI - [Decreased fetal movements and massive fetomaternal transfusion]. PMID- 15544116 TI - [Serious infections after treatment with infliximab]. PMID- 15544117 TI - [Gingival hyperplasia and calcium antagonists]. PMID- 15544118 TI - [Malignant epitheloid nerve sheath tumor]. PMID- 15544119 TI - [Urethral catheterization]. PMID- 15544120 TI - [Picture of the month: pleural effusion]. PMID- 15544121 TI - [Albumin or physiologic saline to hypovolemic intensive care patients?]. PMID- 15544122 TI - [Saline versus Albumin Fluid Evaluation (SAFE) study. Albumin does not increase the mortality of intensive care patients. A study with many aspects]. PMID- 15544123 TI - [Diabetic ketoacidosis versus alcoholic ketoacidosis]. PMID- 15544124 TI - [Vitamin D deficiency]. PMID- 15544125 TI - [Rehabilitation! Diagnosis can be a turning point for adults with ADHD/autism spectrum disorders]. PMID- 15544126 TI - [Adult psychiatry does not recognize child neuropsychiatric disorders. A registry study shows discrepancy between expected and real number of cases]. AB - To identify the group of patients with presumed special needs in adult psychiatry, a list survey was made covering 11 months of diagnostic work in adult psychiatry with a catchment area of 213,000 inhabitants. The survey was made in the urban setting of Gothenburg. During the period of observation, 6,386 patients were diagnosed. Among them 156 (2.4%) were diagnosed with a neuropsychiatric diagnosis of attention deficit hyperactivity disorder, autism spectrum disorders, disorders associated with Tourette's syndrome or mental retardation. The adult psychiatric hospital care does not recognize child neuropsychiatric disorders in the patient population by the frequency the disorders presumably appear. This is especially clear in the group of patients with a combination of narcotic substance abuse and psychiatric symptoms. PMID- 15544127 TI - [New pedagogic activities in medical education. Students actively participate in reorganization of the education]. PMID- 15544128 TI - [The EDIT project. Problem-based learning on the web challenges students' thinking]. AB - EDIT is short for Educational Development using Information Technology. The EDIT project was initiated by the Faculty of Health Sciences at Linkoping University. The aim was to develop web-based scenarios for problem-based learning (PBL). Patient case studies and other medical problems or situations are illustrated by using realistic texts and multimedia, e.g. pictures and short films. Since the project started in 2001, EDIT-scenarios have been developed for six undergraduate programmes. The project covers two years of the medical programme. The use of multimedia has introduced new possibilities to challenge students' thinking by stimulating more senses. The different parts of a scenario are designed to raise questions without providing answers. Both students and tutors have perceived EDIT scenarios as more motivating and interesting than the case studies on paper previously used. EDIT has contributed to improving and updating PBL-scenarios. This process has helped vitalise the discussions about pedagogical issues. For students as well as teachers, the project has lead to increased general familiarity with IT. In this paper, the project concept, practical aspects of the implementation, and pedagogical outcomes are discussed. PMID- 15544129 TI - [Standardized patients--good help in teaching. They function as stand-ins in training of consultation and clinical skills]. PMID- 15544130 TI - [The lower LDL cholesterol the better!]. PMID- 15544131 TI - [Thyrotoxicosis]. PMID- 15544132 TI - [Better routines for the performance of physical examinations by students are required by the MSF]. PMID- 15544133 TI - [Examination in venous thromboembolism: usually ultrasound, but consult guidelines by the National Board of Health and Welfare and do not forget veins of the lower leg]. PMID- 15544134 TI - [Concerning vulvoplasty: sick to treat healthy organs]. PMID- 15544135 TI - [Reply to comments on vulvovaginal plastic surgery: patients' needs and wishes are decisive for our therapeutic strategy]. PMID- 15544136 TI - [Final reply: poor scientific report and reprehensible activity]. PMID- 15544137 TI - [Documented advantages of Lantus in comparison with NPH]. PMID- 15544138 TI - [Sick listing and system errors]. PMID- 15544139 TI - [Patient with sarcoidosis and Lofgren syndrome: can the inflammatory reaction be physiologically favourable for healing?]. PMID- 15544140 TI - Laying the foundations for a transcultural biotechnology law. PMID- 15544141 TI - Society and human genome. PMID- 15544142 TI - [Research with human embryo stem cells. Foundations and judicial limits]. AB - Research with human embryos, and particularly, the use for scientific purposes of human embryonic stem cells has given raise to different sort of problems at the international level. One of the most strict regulation in this field, is this lecture Professors Albin Eser and Hans-Georg Koch analyse the german legal framework in relation with the use of embryos and human embryonic stem cells for scientific purposes. PMID- 15544143 TI - [The legal concept of personhood in Colombia: are there biological reasons to change it?]. AB - Nowadays, as a consequence of scientific advances in embryology, genetics and assisted reproduction techniques, the scientific community ask for new laws to establish new parameters to use with these recent discoveries. Discoveries that have changed the legal concept of "person". In many countries this concept is absolutely useless when facing new techniques such as cloning, stem cell research and embryo storage. That is why it is necessary to change its definition to guarantee the right to life and give a better protection to human embryos. PMID- 15544144 TI - [Assisted reproduction and artificial insemination and genetic manipulation in the Criminal Code of the Federal District, Mexico]. AB - The article that one presents has for purpose outline and comment on the recent modifications to the Penal Code for the Federal District of Mexico which establish, for the first time, crimes related to the artificial procreation and to the genetic manipulation. Also one refers to the interaction of the new legal texts with the sanitary legislation of the country. Since it will be stated in some cases they present confrontations between the penal and the sanitary reglamentation and some points related to the legality or unlawfulness of a conduct that stayed without the enough development. These lacks will complicate the application of the new rules of the Penal Code of the Federal District. PMID- 15544145 TI - [Expansive evolution in the French regulation on genetic fingerprint files after the recent reforms (Part II)]. PMID- 15544146 TI - Prevention of the xenogenic infection risk and the Spanish and German Constitutions. PMID- 15544147 TI - The banking of embryonic stem cells: the legal and ethical framework in the UK. PMID- 15544148 TI - [The scope of the right to life as it relates to the conception according to the European Court of Human Rights]. PMID- 15544149 TI - [Experimentation with extra embryos in Spain: comment on the 45/2003 Law that modifies the 35/1988 Law on assisted reproduction techniques]. AB - The 45/2003 Spanish Act tries to solve the problem of the cryoconserved human embryos produced in in vitro fertilization (IVF) programmes. The new law, which modifies the 35/1988 Act on Artificial Reproduction Techniques, is analyzed from the scientific, ethical and legal points of view. PMID- 15544150 TI - Contrasting expectations of biotechnology for medical care in Taiwan between seniors and medical students. PMID- 15544151 TI - Umbilical cord blood: banking and clinical application. PMID- 15544152 TI - [Human susceptibility to complex diseases. Genetic analysis in large populations]. AB - Inherited susceptibility underlying the ethiopathogenity of complex diseases is based on the existence of a high number of genetic variants which are cooperating to generate different risk genetic profiles. We try to identify such profiles and construct wide data bases with this genetic information. PMID- 15544153 TI - [25 recommendations on the ethical, judicial, and social impact of genetic tests. Brussels 2004]. PMID- 15544154 TI - Awareness of the past, persistence in the present, hope for the future. PMID- 15544155 TI - 2004 general elections--a physician's guide. PMID- 15544156 TI - J. Gregory Cooper, MD, KMA president 2004-2005. Interview by Sharon Heckel. PMID- 15544157 TI - Pediatric cancer in Kentucky: good news. AB - Pediatric cancer is one of the more poignant aspects associated with the occurrence of this prominent chronic disease. Nationally, pediatric cancers occur with vastly lesser frequency than does adult disease, about 1:40 ratio. Nationally the rate is 14.6 per 100,000 population, age, 0-14 years. In Kentucky, all pediatric cancer rates are consistently lower, a statewide rate of 11.3 per 100,000. A rigorous examination was made for any evidence of unusual risk for pediatric cancer within the state; none was found. The population center of Jefferson County was examined with a bit greater detail, owing simply to the larger number. Again, no basis for public health or follow-back was identified. Among the most frequent pediatric cancers, leukemia, central nervous system, and brain were studied for implications of residential proximity to environmental hazards; no evidence of increased risk was identified. In all of the state, only Hardin County evinced any excess pediatric cancer risk, i.e., was significantly greater than the national rates. Yet this highly mobile population [owing to a large military population] may simply represent rates that are more compatible with those of the nation. For the Kentucky comprehensive disease control efforts, then, no emphasis need be placed with pediatric cancer beyond the intense personal tragedy that it poses. PMID- 15544158 TI - Antidepressants--do they cause suicide? PMID- 15544159 TI - Practice what you preach. PMID- 15544160 TI - Stress under the dam: meeting report of the Fourth International Workshop on the Molecular Biology of Stress Responses. PMID- 15544161 TI - Hsp70 mutant proteins modulate additional apoptotic pathways and improve cell survival. AB - Although wild-type Hsp70 (Hsp70WT) inhibits procaspase-3 processing by preventing apoptosome formation, Hsp70WT does not block active caspase-3. Because all caspase-3 inhibitors bear canonical DXXD caspase-3 recognition motifs, we determined whether mutated Hsp70s with caspase-binding motifs act as direct caspase-3 inhibitors. Based on Hsp70 molecular modeling, the DNQP, DEVQ, and EEVD regions localized on the surface of Hsp70WT were chosen, allowing us to design mutants while trying to avoid disrupting the global fold of the molecule and losing the possibility of protein-protein interactions. We replaced DNQP with DQMD, and DEVQ and EEVD with DEVD residues that should be optimal substrates for caspase-3. The resultant Hsp70 mutants directly interacted with active caspase-3 and blocked its proteolytic activity while retaining the ability to reverse protein denaturation and disrupt the interaction between Apaf-1 and procaspase-9. The Hsp70C-terminal mutants interacted with Apaf-1 and active caspase-3 significantly longer than Hsp70WT. The Hsp70 DXXD mutants protected neuron and teratocarcinoma (NT) cells against cell death much better than Hsp70WT whether given before or after serum withdrawal. Hsp70 mutants represent a possible approach to antiapoptotic biotherapeutics. Similar rational designs could be used to engineer inhibitors of additional caspase family members. PMID- 15544162 TI - Intronic hormone response elements mediate regulation of FKBP5 by progestins and glucocorticoids. AB - Expression of FKBP51, a large molecular weight immunophilin, is strongly enhanced by glucocorticoids, progestins, and androgens. However, the activity of a 3.4-kb fragment of the FKBP51 gene (FKBP5) promoter was only weakly increased by progestin and we show here that it is unresponsive to glucocorticoids and androgens. The entire FKBP5 was scanned for consensus hormone response elements (HREs) using MatInspector. We found that 2 regions of intron E, which are conserved in rat and mouse FKBP5, contain HRE-like sequences with high match scores. Deoxyribonucleic acid fragments (approximately 1 kb in length) containing these regions were amplified and tested in reporter gene assays for steroid responsiveness. One region of intron E of FKBP5 (pIE2) conferred both glucocorticoid and progestin responsiveness to 2 heterologous reporter genes, whereas the other, less-conserved region of intron E (pIE1) was responsive only to progestins. The inclusion of pIE1 upstream of pIE2 (pIE1IE2) enhanced progestin but not glucocorticoid responsiveness. None of the constructs containing intronic sequences was responsive to androgens. Mutation of the putative HREs within pIE1 and pIE2 eliminated hormone responsiveness. Electrophoretic mobility shift assays demonstrated that progesterone receptors (PR) bound to the HRE in pIE1, whereas both PR and glucocorticoid receptors interacted with the HRE in pIE2. These data suggest that distal intronic elements significantly contribute to transcriptional regulation of FKBP5 by glucocorticoids and progestins. PMID- 15544163 TI - Localization and function in endoplasmic reticulum stress tolerance of ERdj3, a new member of Hsp40 family protein. AB - Heat shock protein 40 (Hsp40) family proteins are known to bind to Hsp70 through their J-domain and regulate the function of Hsp70 by stimulating its adenosine triphosphatase activity. In the endoplasmic reticulum (ER), there are 5 Hsp40 family proteins known so far, 3 of which were recently identified. In this report, one of the novel Hsp40 cochaperones, ERdj3, was characterized in terms of its subcellular localization, stress response, and stress tolerance of cells. By using ERdj3-specific polyclonal antibody, endogenous ERdj3 protein was shown to reside in the ER as gene transfer-mediated exogenous ERdj3. Analysis of the expression level of endogenous ERdj3 protein revealed its moderate induction in response to various ER stressors, indicating its possible action as a stress protein in the ER. Subsequently, we analyzed whether this molecule was involved in ER stress tolerance of cells, as was the case with the ER-resident Hsp70 family protein BiP. Although overexpression of ERdj3 by gene transfection could not strengthen ER stress tolerance of neuroblastoma cells, reduction of ERdj3 expression by small interfering ribonucleic acid decreased the tolerance of cells, indicating that ERdj3 might have just a marginal role in the ER stress resistance of neuroblastoma cells. In contrast, overexpression of ERdj3 notably suppressed vero toxin-induced cell death. These data suggest that ERdj3 might have diverse roles in the ER, including that of the molecular cochaperone of BiP and an as yet unknown protective action against vero toxin. PMID- 15544164 TI - Downstream caspases are novel targets for the antiapoptotic activity of the molecular chaperone hsp70. AB - The response of cancer cells to apoptosis-inducing agents can be characterized by 2 opposing factors, the proapoptotic caspase cascade and the antiapoptotic stress protein Hsp70. We show here that these factors interact in U-937 leukemia cells induced to apoptosis with anticancer drugs, etoposide and adriamycin (ADR). The protective effect of Hsp70 was verified using 2 approaches: mild heat stress and transfection-mediated overexpression of the Hsp70 gene. The increase in Hsp70 levels attained by these 2 methods was found to postpone caspase activation for 12-18 hours. An in vitro assay was developed using mouse myeloma NS0/1 cells, which lack the expression of Hsp70. Measurement of DEVD-ase activity in extracts of apoptotic NS0/1 cells incubated with purified Hsp70 showed that Hsp70 reduced caspase activity by up to 50% of its control value in a dose-dependent manner. The hypothesis that the inhibitory effect of Hsp70 on caspase-3/7 activity related to a direct interaction between Hsp70 and the caspases was tested by reciprocal immunoprecipitations and Far-western analyses. These tests were performed with extracts of Hsp70-overexpressing, control, and ADR-treated U-937 cells and using anti-caspase-3, caspase-7, and anti-Hsp70 antibodies, and the data clearly showed that Hsp70 was able to interact with the proforms of these caspases in cell lysates and with reconstituted purified proteins but did not bind the activated forms of either caspase-3 or -7. This association was also corroborated by a novel, enzyme-linked immunosorbent assay-like assay, protein interaction assay, that combined the advantages of immunoprecipitation and immunoblotting in a 96-well microplate-based assay. Thus, Hsp70 may act to suppress caspase-dependent apoptotic signaling through binding the precursor forms of both caspase-3 and caspase-7 and preventing their maturation. PMID- 15544165 TI - Exercise induces the release of heat shock protein 72 from the human brain in vivo. AB - The present study tested the hypothesis that in response to physical stress the human brain has the capacity to release heat shock protein 72 (Hsp72) in vivo. Therefore, 6 humans (males) cycled for 180 minutes at 60% of their maximal oxygen uptake, and the cerebral Hsp72 response was determined on the basis of the internal jugular venous to arterial difference and global cerebral blood flow. At rest, there was a net balance of Hsp72 across the brain, but after 180 minutes of exercise, we were able to detect the release of Hsp72 from the brain (335 +/- 182 ng/min). However, large individual differences were observed as 3 of the 6 subjects had a marked increase in the release of Hsp72, whereas exercise had little effect on the cerebral Hsp72 balance in the remaining 3 subjects. Given that cerebral blood flow was unchanged during exercise compared with values obtained at rest, it is unlikely that the cerebral Hsp72 release relates to necrosis of specific cells within the brain. These data demonstrate that the human brain is able to release Hsp72 in vivo in response to a physical stressor such as exercise. Further study is required to determine the biological significance of these observations. PMID- 15544166 TI - Mitochondrial mechanism of heat stress-induced injury in rat cardiomyocyte. AB - Heat stress results in cardiac dysfunction and even cardiac failure. To elucidate the cellular and molecular mechanism of cardiomyocyte injury induced by heat stress, the changes of structure and function in cardiac mitochondria of heat exposed Wistar rats and its role in cardiomyocyte injury were investigated. Heat stress induced apoptosis and necrosis of cardiomyocytes in a time- and dose dependent fashion. In the mitochondria of heat-stressed cardiomyocytes, the respiratory control rate and oxidative phosphorylation efficiency (P:O) were decreased gradually with the rise of rectal temperature. The Ca2+ -adenosine triphosphatase activity and Ca2+ content were also reduced. Exposing isolated mitochondria to the heat stress induced special internal environmental states including Ca2+ overload, oxidative stress, and altered mitochondrial membrane permeability transition (MPT). In vivo, the heat stress-induced mitochondrial MPT alteration was also found. The changes of mitochondrial MPT resulted in the release of cytochrome c from mitochondria into the cytosol, and in turn, caspase 3 was activated. Transfection of bcl-2 caused Bcl-2 overexpression in cardiomyocyte, which protected the mitochondria and reduced the heat stress induced cardiomyocyte injury. In conclusion, it appears that the destruction of mitochondrial structure and function not only resulted in the impairment of physiological function of cardiomyocytes under heat stress but may also further lead to severe cellular injury and even cell death. These findings underline the contribution of mitochondria to the injury process in cardiomyocytes under heat stress. PMID- 15544167 TI - The heat shock protein HSP70 and heat shock cognate protein HSC70 contribute to antimony tolerance in the protozoan parasite leishmania. AB - Antimony-containing drugs are still the drugs of choice in the treatment of infections caused by the parasite Leishmania. Resistance to antimony is now common in some parts of the world, and several mechanisms of resistance have been described. By transfecting cosmid banks and selecting with potassium antimonyl tartrate (SbIII), we have isolated a cosmid associated with resistance. This cosmid contains 2 copies of the heat shock protein 70 (HSP70) and 1 copy of the heat shock cognate protein 70 (HSC70). Several data linked HSP70 to antimony response and resistance. First, several Leishmania species, both as promastigotes and amastigotes, increased the expression of their HSP70 proteins when grown in the presence of 1 or 2 times the Effect Concentration 50% of SbIII. In several mutants selected for resistance to either SbIII or to the related metal arsenite, the HSP70 proteins were found to be overexpressed. This increase was also observed in revertant cells grown for several passages in the absence of SbIII, suggesting that this increased production of HSP70 is stable. Transfection of HSP70 or HSC70 in Leishmania cells does not confer resistance directly, though these transfectants were better able to tolerate a shock with SbIII. Our results are consistent with HSP70 and HSC70 being a first line of defense against SbIII until more specific and efficient resistance mechanisms take over. PMID- 15544168 TI - Attenuation of okadaic acid-induced hyperphosphorylation of cytoskeletal proteins by heat preconditioning and its possible underlying mechanisms. AB - An imbalanced phosphorylation system is recognized to be one of the main reasons for Alzheimer-like hyperphosphorylation of cytoskeletal proteins. However, little is known about the strategies rectifying the lesions caused by this disrupted phosphorylation. To search for the means to arrest Alzheimer-like damages and explore the underlying mechanisms, in this study we treated N2a/peuht40 cells with okadaic acid (OA), a specific inhibitor of protein phosphatase-2A (PP-2A) and PP-1, to mimic an Alzheimer-like phosphatase-deficient system and then used heat preconditioning (42 degrees C for 1 hour) to induce the expression of inducible heat shock protein 70 (Hsp70) in the cells. We observed that heat preconditioning arrested OA-induced hyperphosphorylation of neurofilament (NF) protein at SMI34 and SMI33 epitopes as well as hyperphosphorylation of tau at Tau 1 and PHF-1 epitopes. It counteracted OA-induced decrease in PP-2A activity with a concurrent inhibition in constitutive activity of mitogen-activated protein kinases (MAPKs) and cyclic adenosine 5'-monophosphate-dependent protein kinase A (PKA). Conversely, quercetin, a recognized blocker of stress-responsive Hsp70 expression, diminished the effects caused by heat preconditioning. These results suggested that Hsp70 antagonized OA-induced Alzheimer-like NF and tau hyperphosphorylation, and the restoration of PP-2A and inhibition of MAPKs-PKA activity might be part of the underlying mechanisms for the rectification of OA induced hyperphosphorylation. PMID- 15544170 TI - A quality initiative--can we reduce the incidence of hypotension during hemodialysis? AB - "Dialysis-related side effects have been identified as an important cause of reduced dialysis efficiency" (Donauer, Kolblin, Bek, Krause & Bohler, 2000, p. 115). Certain patients experience symptomatic hypotension with hemodialysis treatments, adding to patient discomfort and morbidity, and preventing optimal fluid removal. These symptoms require prompt nursing interventions to mitigate potentially severe complications. Technology today allows for continuous non invasive monitoring of blood volume during the hemodialysis treatment, automatic measurement and recording of blood pressure, and the use of sodium modelling and ultrafiltration (UF) profiles. As a teaching strategy, as well as a quality improvement initiative, a task force of volunteer staff nurses was established to monitor selected patients, utilizing this technology, in an effort to reduce episodes of hypotension and improve patient outcomes. A special project team was recruited, in-services on blood volume monitoring, sodium and UF profiling were held for all staff, and tools to assess the current status of the patient and monitor progress were designed and implemented. Patients who were followed reported feeling better and experienced fewer hypotensive episodes. These outcomes are likely a result of several factors including: intensive monitoring by a consistent group, review and analysis of dialysis records every two weeks, implementation of several interventions (such as instituting sodium ramping, changing UF profiles, changing antihypertensive medication, and/or patient education), and evaluating these changes within two weeks. PMID- 15544171 TI - Highlights of the 2003 Clinical Practice Guidelines for the prevention and management of diabetes in Canada. PMID- 15544169 TI - Molecular cloning and expression of two HSP70 genes in the prawn, Macrobrachium rosenbergii. AB - Two complementary deoxyribonucleic acid (cDNA) clones encoding 2 different 70-kDa heat shock proteins (HSPs) were isolated from the prawn Macrobrachium rosenbergii. The cDNA clones were 2448 and 2173 bp in length and contained 1950- and 1734-bp open reading frames (ORFs), respectively. The ORFs encoded 649- and 577-amino acid polypeptides, which were named Mar-HSC70 and Mar-HSP70, respectively, according to the sequence identities with other known HSC70s and HSP70s and based on their inducibility in response to heat shock stress (at 35 degrees C). Genomic DNA sequence analysis revealed no introns in either gene. The major structural differences between the 2 proteins were a 60-amino acid segment and a 14-amino acid segment present in the N-terminal and C-terminal, respectively, of Mar-HSC70 that were not found in Mar-HSP70. Northern blotting and semiquantitative reverse transcription-polymerase chain reaction analyses indicated that the Mar-HSP70 gene was expressed under heat shock (35 degrees C) stress in a non-tissue-specific manner. In contrast, Mar-HSC70 messenger ribonucleic acid was constitutively expressed in every tissue except muscle, and its expression in response to heat shock (at 35 degrees C) changed only in muscle. PMID- 15544172 TI - Of loss and honour. PMID- 15544173 TI - Treatment of acute gouty arthritis in patients with chronic kidney disease. PMID- 15544174 TI - Computer-assisted learning. PMID- 15544175 TI - Is it time to re-examine structure of your quality department? AB - --Periodically review the structure that governs your QI efforts. --The search for maximum efficiency and benefit is one of the leading causes for re examination. --Make sure the right people are made accountable for key quality decisions. PMID- 15544176 TI - Program enables 50 new initiatives in four months: 'Transforming Care at the Bedside' program. AB - Program targets bedside care on a standard hospital medical or surgical unit. Each staff nurse took on several projects, and all progressed simultaneously. Post-op order sets were adopted by nearly 100% of OB/GYN and orthopedic surgeons. PMID- 15544177 TI - Research to practice is a hard journey, experts say. AB - --Conference brings together researchers and users to address greatest challenges. --Presenting information at several different levels increases chances of practice improvement. --Networks of delivery systems jointly pursue studies, and shared research results. PMID- 15544178 TI - JCAHO resource compares more than 16,000 facilities. AB - --Proponents seek to take the guesswork out of choosing a hospital for consumers. --Facilities' care compared in heart attack, heart failure, pneumonia, and pregnancy-related conditions. --National quality goals, and patient safety goals are incorporated into evaluation process. PMID- 15544179 TI - Study shows 12-hour shifts increase errors. AB - --Anonymity was key in ensuring candid reportinc of errors and near-errors. --The likelihood of making an error increases as the total number of hours worked increases. --The data are to be used for pilot study to test a fatigue countermeasures program. PMID- 15544181 TI - Patient safety alert. 'Safety culture' approach guides health system's efforts. PMID- 15544180 TI - JCAHO unveils national patient safety goals. PMID- 15544182 TI - "Each one, teach one'. PMID- 15544183 TI - Racism and the NHS. PMID- 15544184 TI - Lesson from the law. PMID- 15544185 TI - Six steps to power. PMID- 15544186 TI - Stroke of luck. Interview by Lucy Gooding. PMID- 15544187 TI - More than survival. PMID- 15544188 TI - The research agenda for protocol-based care. AB - BACKGROUND: Protocol-based care is increasingly being used to deliver collaborative, integrated and improved patient-centred care, based on the best available evidence. This article provides an overview of key issues arising from protocol-based care literature to illustrate the research agenda for this important care delivery approach. CONCLUSION: Protocols provide great potential to deliver best practice. However, questions remain about the benefits of protocol-based care. PMID- 15544189 TI - The new childhood immunisation schedule. PMID- 15544190 TI - Sudden bereavement in acute care settings. AB - AIM: This literature review examines best practice in caring for those who have been bereaved suddenly in acute care settings. Theories of bereavement are outlined in relation to sudden death. The evidence base for best practice is presented and the role of the nurse in this situation is examined. CONCLUSION: There are common psychological, physical and behavioural manifestations of grief, but people experiencing the sudden death of a loved one are at risk of more pronounced and prolonged grief reactions than those who had been expecting death. Nurses have an important role in facilitating bereavement in acute care settings. Preparation for bereavement care begins before an individual has died and continues through to identifying the appropriate person to provide follow-up care. Those who are unexpectedly and suddenly bereaved should receive practical and considerate evidence-based care. PMID- 15544191 TI - Using the Internet to conduct a literature search. AB - This article provides advice on how to carry out a successful literature search using electronic databases available on the Internet. PMID- 15544192 TI - Listen and learn. PMID- 15544193 TI - Taking counsel. PMID- 15544194 TI - Mean body weight, height, and body mass index, United States 1960-2002. AB - This report presents trends in national estimates of mean weight, height, and body mass index (BMI) from the National Health Examination and the National Health and Nutrition Examination Surveys between 1960 and 2002. The tables included in this report present data for adults by sex, race/ethnicity, and age group and for children by sex and year of age. Mean weight and BMI have increased for both sexes, all race/ethnic groups, and all ages. Among adults, mean weight increased more than 24 pounds. Although not as dramatically, mean height has also increased for most ages and for both males and females. PMID- 15544195 TI - What's news in pediatric dermatology: an update. PMID- 15544196 TI - Warts and molluscum in children. PMID- 15544197 TI - Neurofibromatosis-1 in childhood. PMID- 15544198 TI - Pediatric dermatologic surgery: a surgical approach to the cutaneous features of tuberous sclerosis complex. PMID- 15544199 TI - Surgical repair of the auricle. AB - Secondary intention, primary closure, and full thickness skin grafts can handle the majority of ear closures. Transposition flaps work nicely at the root of the helix, the preauricular area, the intertragal notch and the postauricular area. Helical rim advancements and their variations are the workhorse for repairing and restoring the natural arch of the helix. Retroauricular 2-stage pedicle flaps with or without a cartilage graft will provide a nice cosmetic result for larger defects involving the helical rim. Remember, most importantly, know your wound, know your patient, and the simplest closure is often the best one. PMID- 15544200 TI - Vascular endothelial cell adhesion and signaling during leukocyte recruitment. AB - During inflammation, coordinated expression of cytokine-induced adhesion molecules (CAMs) on postcapillary venular endothelial cells (ECs) regulates leukocyte recruitment. During their recruitment from blood, leukocytes adhere to EC CAMs, activating signaling pathways inside ECs. In a forthcoming paradigm, leukocyte transendothelial migration requires active EC participation, with extracellular adhesive CAM functions mirrored by cytoplasmic do-main-dependent intracellular events. These events serve to reorganize the EC actin cytoskeleton. Investigators have visualized this as changes in EC shape, transient opening of EC-EC contacts, and redistribution of CAMs expressed on the luminal EC surface. In this review, we (1) summarize the overlapping extracellular adhesive properties of the 3 EC CAMs most important for leukocyte recruitment during inflammation, namely, E-selectin, vascular cell adhesion molecule, and intercellular adhesion molecule-1; (2) explore the role of these 3 CAMs as signal transducers by identifying the intracellular signals (Ca++, Rho/Rac, and phosphatidylinositol 4,5-bisphosphate) that upon leukocyte engagement, reorganize the EC cytoskeleton and redistribute these apical CAMs, thereby favoring leukocyte recruitment; and (3) describe how CAM-derived signals lead to ezrin radixin-moesin complex formation and how this complex of plasma membrane cytoskeleton adapter proteins coordinates CAM-driven intracellular signals with extracellular adhesive CAM functions. This literature review suggests that the cytoplasmic domains of these EC CAMs and their downstream effectors represent new and potentially beneficial intracellular therapeutic targets for treating diseases of the skin. PMID- 15544201 TI - Cutaneous biology of gammadelta T cells. AB - The vast majority of serious noninfectious skin diseases are the result of inappropriate inflammatory responses (eg, atopic dermatitis, psoriasis) or neoplastic transformation (eg, squamous cell carcinoma, melanoma, cutaneous T cell lymphoma). T cells are critical to both of these major disease categories, playing critical roles in driving or controlling inflammation, as well as the immunosurveillance of cutaneous tumors. T cells may express either 1 of 2 major T cell receptor (TCR) subtypes composed of heterodimeric alphabeta or gammadelta proteins. While most T cells in the blood and lymph nodes are of the alphabeta type and demonstrate vast heterogeneity, gammadelta T cells are relatively enriched in epithelial tissues and often show less TCR variability. This distinction is especially evident in mice and several other mammals, as well as in humans, albeit to a lesser degree. Recent studies in laboratory animals have indicated the capacity of gammadelta T cells to play major roles in maintenance of the epidermal barrier, regulation of cutaneous inflammation, and protection against cutaneous neoplasms. This review will expound on the biology of gammadelta T cells, their relationship to the skin, and the implications for our understanding of cutaneous disease. PMID- 15544202 TI - Advances in understanding the immunobiology and immunotherapy of cutaneous T-cell lymphoma. PMID- 15544203 TI - Contact allergens of the year. PMID- 15544204 TI - Evidence-based evaluation of immunomodulatory therapy for the cutaneous manifestations of lupus. AB - This literature review revealed an unexpected paucity of data derived from adequately powered, randomized, controlled trials to guide evidence-based treatment of the cutaneous manifestations of lupus. Lupus erythematosus is characterized by spontaneous fluctuations in disease activity. This characteristic may serve to explain some of the contradictions in published evidence and cautions us not to rely too heavily on data derived from inadequately powered studies, small uncontrolled case series, or individual case reports, which are additionally subject to publication bias. Clearly, there is a pressing need to conduct adequately powered clinical research studies to fill these gaps in our knowledge. There also is a need for controlled trials to assess drug safety in patients with SLE. Much of the data regarding side-effect profiles of the immunomodulatory drugs has been compiled in patients receiving organ transplantation or chemotherapy for malignant diseases and extrapolated to patients with lupus. Given that there is often a substantial overlap in symptoms related to drug toxicity and to lupus erythematosus, careful studies should be conducted to assess the safety and toxicities of both established and newer medical therapies, especially those whose efficacy is supported only by small trials and case reports. Table 4 summarizes the results of our literature review. For each drug, we give the highest level of data we found, both for the treatment of lupus erythematosus in general and for the treatment of the cutaneous manifestation of lupus in particular. PMID- 15544205 TI - Innovative treatments for chronic dermatitis. PMID- 15544206 TI - Advances in prognostication of cutaneous malignant melanoma. PMID- 15544207 TI - Update in the diagnosis of keratinocytic neoplasia. PMID- 15544208 TI - In vivo reflectance-mode confocal scanning laser microscopy in dermatology. AB - RCM offers tremendous potential for the advancement of medical research and clinical care. In research, it offers benefits both ex vivo and in vivo. Ex vivo, it can allow us to sample tissue and evaluate it noninvasively to determine what further testing--on the same exact tissue--may be helpful. In vivo RCM can be used to study normal or pathophysiologic processes in real-time noninvasively and by the same technique sequentially over time. Immunologic events previously only studied ex vivo or by static images can be traced from their inception to completion (Table 1). The potential of RCM in vivo is tremendous. How would our world change if we could noninvasively diagnose skin lesions and, with the advent of new minimally invasive therapies, administer treatment and noninvasively monitor that treatment? The potential to allow better medical care based on actual visualization of therapeutic response and healing is obvious. Much like early X-ray and ultrasound imaging, RCM is in its infancy. It is only a matter of time and continued persistent research that will lead to similar success and utility for RCM. PMID- 15544209 TI - Prepare for greatness. PMID- 15544210 TI - Reported adverse effects and drug interactions of triazolam (Halcion). PMID- 15544211 TI - The preparation and modification of study models for esthetic restorative cases. PMID- 15544212 TI - Emergency situations. PMID- 15544213 TI - Nutrition, diet, and dentistry today. PMID- 15544214 TI - Marginal gap repair with flowable resin-based composites. AB - This study was designed to test the hypothesis that the flowable/dentin interface has the weakest bond and highest frequency of failure when a resin-based composite is repaired using a flowable composite. Comparing three specific groups -dentin, flowable composite, and hybrid composite-under the Kruskal-Wallis test showed that the mean bond strength values at the flowable/dentin interface and within the dentin were significantly lower than those at the hybrid/flowable interface and within the flowable resin. Dentin near the pulp chamber yielded consistently lower bond strength values than dentin located at the dentino-enamel junction. Bonding at the flowable resin/dentin interface showed the weakest bond and highest frequency of failure. PMID- 15544216 TI - Adding the closed mouth impression technique to a cast impression coping technique. AB - The closed mouth impression technique is a well-accepted technique for constructing single crowns in an individual quadrant. This article presents two cases in which the cast coping technique was used to create a more stable impression of the preparation itself. PMID- 15544215 TI - Laser-assisted new attachment procedure in private practice. AB - Three private dental practices conducted a retrospective analysis of patients receiving the laser-assisted new attachment procedure (LANAP). Retrospective results were compared to clinical trial data from the University of Texas Health Sciences Center in San Antonio (UTHSCSA) to determine if outcomes from a controlled clinical trial can be duplicated in private practice. Results also are compared with published results of other surgical and nonsurgical therapies for inflammatory periodontal disease. PMID- 15544217 TI - Rare and abnormal massive dental calculus deposit: an investigative report. AB - An unprecedented case is presented involving a massive calculus buildup on the mandibular incisors. The clinical and radiographic findings were reviewed, probable causes were investigated, and results were outlined and discussed. The composition, origin, and formation of dental calculus and its interrelationship with dental plaque and saliva were highlighted. The role of dental calculus in the pathogenicity of periodontal diseases, in view of epidemiological data, research, and clinical study findings, is discussed. Both the case management and the benefit of total calculus removal for resolving periodontal disease are underlined. PMID- 15544218 TI - Orthodontic extrusion anchored in osseointegrated implants: a case report. AB - Rapid orthodontic extrusion is indicated for cases involving biological space invasion of the periodontal ligament in which the surgical increase of the clinical crown can compromise esthetics or the support of the adjacent teeth. This article presents the case report of a tooth with radicular perforation 1.0 mm below the bone crest. A procedure to restore the biological distances was necessary. Rapid orthodontic extrusion of the affected tooth, with anchorage in the adjacent osseointegrated implants, was selected. PMID- 15544219 TI - A study of 55 submandibular salivary gland excisions. AB - This study examines 55 cases involving patients who underwent submandibular gland excision. Their preoperative diagnostic examinations, postoperative complications, and glandular pathology are discussed. Sialolithiasis was the main histopathological finding, followed by chronic sialadenitis. Temporary paresis of the mandibular branch of the facial nerve was the most common postoperative complication. Removing the submandibular salivary gland may be followed by a number of complications, most of which can be avoided if the appropriate surgical technique is applied. PMID- 15544220 TI - Superficial mucocele of the labial mucosa: a case report and review of the literature. AB - Superficial mucocele is considered a relatively common, uncomplicated, and truly benign oral lesion, although a systematic review of the literature revealed only 27 well-documented cases. The general dentist frequently is confronted with questions from patients regarding these often recurrent blisters. While trauma is suspected to be the cause, the etiology of these lesions is not understood clearly due to the insufficient number of reported cases; as a result, their optimal management remains in question. Insufficient knowledge of the clinical appearance and histopathologic features of this lesion may generate diagnostic confusion, leading to improper diagnosis and inadequate management. This article presents an atypical case of superficial mucocele and summarizes the findings of all previously reported cases to emphasize the variable clinical features and increase general dentists' knowledge of the clinical spectrum regarding this condition's signs and symptoms. PMID- 15544221 TI - Stress among dentists. AB - Dentists are faced every day with many stressors; as a result, they are subjected to many symptoms of stress that must be identified and managed in the early stages before serious physical and psychological consequences develop. This article reviews the literature concerning stress among dentists. Many stressors are identified and coping techniques are explained. PMID- 15544222 TI - Mucoepidermoid carcinoma in a young adult: recognition, diagnosis, and treatment and responsibility. AB - Recognizing an abnormality is the first and most essential step when diagnosing a pathosis. All health care providers are responsible for evaluating suspicious symptoms and clinical and laboratory findings. Without a baseline evaluation, dentists cannot guarantee that patients are free of potentially serious disease. This article documents how the prognosis for and morbidity of cancer therapy may have been affected by the failure to recognize abnormal signs. Professional responsibility and liability for recognition of abnormality and subsequent diagnosis is emphasized. PMID- 15544223 TI - Dental implications of mood disorders. AB - Mood disorders refers to a heterogenous group of mental conditions characterized by extreme exaggeration and disturbance of mood and affect. This article examines major depression and bipolar disorders and how these disorders can affect a patient's dental care. The drugs used to treat these conditions have significant side effects that dentists should be aware of. In addition, important drug interactions can occur as a result of agents used by the dentist. The dentist should refer patients found with signs and symptoms of mood disorders for medical evaluation and treatment. Depressed patients often have poor oral hygiene due to a lack of interest in self-care. Xerostomia occurs due to depression and the drugs used to treat it. The dentist needs to provide an aggressive preventive dental education program for these patients, including the use of artificial salivary products, mouthwashes, and topical fluoride applications, in addition to the treatment of candidiasis when present. PMID- 15544224 TI - Esthetic removable partial dentures. AB - This article provides information regarding the many ways that removable partial dentures (RPDs) may be used to solve restorative problems in the esthetic zone without displaying metal components or conspicuous acrylic resin flanges. The esthetic zone is defined and described, as are methods for recording it. Six dental categories are presented that assist the dentist in choosing a variety of RPD design concepts that may be used to avoid metal display while still satisfying basic principles of RPDs. New materials that may be utilized for optimal esthetics are presented and techniques for contouring acrylic resin bases and tinting denture bases are described. PMID- 15544225 TI - Oral Diagnosis: interradicular radiolucency. PMID- 15544226 TI - Oral Diagnosis: mandibular radiolucency with central radiopacity. PMID- 15544227 TI - Interactive haptic modeling of colon and colonoscope. AB - This paper presents physics-based modeling for colonoscopy training simulator. The colon is modeled as a chain of beam along the medial axis for global bending motions. The Timoshenko's beam theory is applied to the centerline of colon extracted from a medical image. The stiffness matrix of colon is formulated using the finite element method. The colonoscope model consists of rigid elements connected with torsional spring and damper. This modeling allows global bending motions to be simulated in real time. PMID- 15544228 TI - Computer-based simulator for catheter insertion training. AB - Minimally invasive surgery procedures are getting common in surgical practice; however the new interventional procedure requires different skills compared to the conventional surgical techniques. The need for training process is very important in order to successfully and safely execute a surgical procedure. Computer-based simulators, with appropriate tactile feedback device, can be an efficient method for facilitating the education and training process. In addition, virtual reality surgical simulators can reduce costs of education and provide realism with regard to tissues behaviour and real-time interaction. This work take into account the results of the HERMES Project (HEmatology Research virtual MEdical System), conceived and managed by Consorzio CETMA-Research Centre; the aim of this project is to build an integrate system in order to simulate a coronary angioplasty intervention. PMID- 15544229 TI - Distributed interactive virtual environments for collaborative experiential learning and training independent of distance over Internet2. AB - Medical knowledge and skills essential for tomorrow's healthcare professionals continue to change faster than ever before creating new demands in medical education. Project TOUCH (Telehealth Outreach for Unified Community Health) has been developing methods to enhance learning by coupling innovations in medical education with advanced technology in high performance computing and next generation Internet2 embedded in virtual reality environments (VRE), artificial intelligence and experiential active learning. Simulations have been used in education and training to allow learners to make mistakes safely in lieu of real life situations, learn from those mistakes and ultimately improve performance by subsequent avoidance of those mistakes. Distributed virtual interactive environments are used over distance to enable learning and participation in dynamic, problem-based, clinical, artificial intelligence rules-based, virtual simulations. The virtual reality patient is programmed to dynamically change over time and respond to the manipulations by the learner. Participants are fully immersed within the VRE platform using a head-mounted display and tracker system. Navigation, locomotion and handling of objects are accomplished using a joy-wand. Distribution is managed via the Internet2 Access Grid using point-to-point or multi-casting connectivity through which the participants can interact. Medical students in Hawaii and New Mexico (NM) participated collaboratively in problem solving and managing of a simulated patient with a closed head injury in VRE; dividing tasks, handing off objects, and functioning as a team. Students stated that opportunities to make mistakes and repeat actions in the VRE were extremely helpful in learning specific principles. VRE created higher performance expectations and some anxiety among VRE users. VRE orientation was adequate but students needed time to adapt and practice in order to improve efficiency. This was also demonstrated successfully between Western Australia and UNM. We successfully demonstrated the ability to fully immerse participants in a distributed virtual environment independent of distance for collaborative team interaction in medical simulation designed for education and training. The ability to make mistakes in a safe environment is well received by students and has a positive impact on their understanding, as well as memory of the principles involved in correcting those mistakes. Bringing people together as virtual teams for interactive experiential learning and collaborative training, independent of distance, provides a platform for distributed "just-in-time" training, performance assessment and credentialing. Further validation is necessary to determine the potential value of the distributed VRE in knowledge transfer, improved future performance and should entail training participants to competence in using these tools. PMID- 15544230 TI - Real time estimation of physical activity and physiological performance reserves of players during a game of soccer. AB - The need for ever increasing achievements in athletic performance is pushing the acquisition costs and salaries of top performers to unprecedented heights. Individual talent, good physical condition and relentless training, however, do not by themselves guarantee success of the team effort. Team performance is determined by game strategy and team coordination. Ideally, during a game, the team coach should be able to gauge the instantaneous physical condition and performance reserves of each player on the team and direct game strategy accordingly. Our experimental setup explores the feasibility of objectively estimating, in real time, the intensity of physical activity and the physiological performance reserves of each and every player on the team during a game of soccer. PMID- 15544231 TI - The Korydallos, Greece, prisons telemedicine system experience: why technology alone is not a sufficient condition. AB - In 1999 the Greek Ministry of Justice decided to utilise telemedicine to improve health services in the largest prison in Greece (Korydallos Prison). The Nikea Hospital in Piraeus undertook to support the effort. For 12 months following installation, intensive "hands-on" training on the use of the system was offered to the staff of both the Korydallos Prison and the Nikea Hospital. However serious operational problems related either to prison bureaucracy or to the inflexibility the Greek National Health Service has annulled the effectiveness of the Korydallos Prison telemedicine system. Still, analysis of the system development history reveals that: (1) if freed from bureaucratic and labour related obstacles, prison telemedicine is a viable option. (2) Telemedicine can avert transfers to out-of-prison medical facilities. (3) If properly implemented, telemedicine can generate substantial savings and (4) telemedicine can greatly improve the quality of care available to prisoners. PMID- 15544232 TI - A clinical and educational telemedicine link between Bulgaria and Greece. AB - During its transition to a free economy, Bulgaria benefited from foreign aid provided by Greece. One of the projects was the clinical and educational telemedicine link between the Medical University of Varna in Bulgaria and the Faculty of Medicine of Aristotle University of Thessaloniki in Greece. This began in 1997. In terms of the educational activities, the Bulgarian side of the network supports (a) electronic design and publishing activities, (b) web hosting and mail server activities and (c) satellite communications. In addition it supports an electronic classroom equipped with personal workstations, multimedia projectors and videoconference facilities. Communications are via the ISDN network. In terms of its telemedicine activities, the network provides remote medical assistance to "language handicapped" travellers and to migrant workers in both countries. The main clinical experience is remote consultations in immunology. This admittedly limited experience demonstrates that telemedicine can be used to provide assistance to remote colleagues. In cases where the patient cannot communicate with the attending physician, the use of telemedicine can greatly improve the quality of care available to travellers and migrant workers. PMID- 15544233 TI - Hybrid-reality: collaborative biomedical data exploration exploiting 2-D and 3-D correspondence. AB - Doctors and radiologists are trained to infer and interpret three-dimensional information from two-dimensional images. Traditionally, they analyze sets of two dimensional images obtained from imaging systems based on x-rays, computed tomography, and magnetic resonance. These images correspond to slices or projections to a single plane. With the resolution of the scanners increasing in all directions, so is the complexity of the data that can be used for diagnostic purposes. Using volume rendering techniques, massive stacks of image slices can be combined into a single image and important features stressed, increasing the doctor's ability to extract useful information from the image. A hybrid visualization approach combining 2D slices and 3D visuals is presented, drawing from the best features of both of these approaches. 2D slices emulate conventional medical images while 3D images provide additional information, such as better spatial location of the features in the surrounding structures as well as the 3D shape of features. PMID- 15544234 TI - Manipulation of volume data to manufacture vascular replicas. AB - There are three reasons to create physical replicas of human anatomy: (1) to be able to better visualize the shape of a single organ, or a section of anatomy; (2) to be able to visualize the spatial relationships in three-dimensions; and (3) to use accurate replicas to practice or rehearse otherwise high-risk clinical procedures in the laboratory. This paper describes a project to fabricate a carotid artery. It discusses the gathering of data, the conversion to a volume, and the subsequent conversion to a manufacturable form. PMID- 15544235 TI - A 3D interactive multimodal viewer as data mining tool for the Visible Human Dataset color image histograms. AB - An on-line virtual three-dimensional immersive environment to navigate through colorimetric characterization of the Visible Human Dataset (VHD) cryosectional cross-section color images is introduced. Real-time analysis of color component characteristics of a user defined set of VHD images is now possible. This is a potentially useful resource to many developers working on the VHD raw data, however it could be used in medical education. PMID- 15544236 TI - An active constraint environment for minimally invasive heart surgery: early experience of a cutting operation. AB - Master/slave telemanipulator systems can be applied in minimally invasive heart surgery. However, due to the beating heart and difficulties of finding inner points inside the heart, a surgical task operation such as cutting can be very difficult. In order to avoid surgical error, the "active constraint" concept can be applied. This paper shows an example of an "active constraint" environment used for minimally invasive heart surgery. Experiments have been carried out for a 2-DOF master and the preliminary results validate the present approach. PMID- 15544237 TI - Quantifying surgeon grasping mechanics in laparoscopy using the Blue DRAGON system. AB - Mechanical testing of abdominal organs has a profound impact on surgical simulation and surgical robotics development. Due to the nonlinear and viscoelastic nature of soft tissue it is crucial to test them in surgically relevant ranges of applied force, deformation, and duration for incorporating haptic realism into surgical simulators and for safe operation of surgical robots. In order to determine these ranges, a system known as the Blue DRAGON was used to track the motions and the forces applied to surgical tools during live procedures for quantifying how surgeons typically perform a minimally invasive surgical procedure. Thirty-one surgeons of varying skill were recorded performing three different surgical tasks. Grasping force (as applied to the tool handles) and handle angle for each tool were the signals of interest among 26 channels total acquired by the system in real time. These data were analyzed for their magnitudes and frequency content. Using the tool contact state, an algorithm selected tissue grasps to analyze measures during grasps only, as well as obtain grasp durations. The mean force applied to the tool handles during tissue grasps was 8.52 N +/- 2.77 N; maximum force was 68.17 N. Ninety-five percent of the handle angle frequency content was below 1.98 Hz +/- 0.98 Hz. Average grasp time was 2.29 s +/- 1.65 s, and 95% of all grasps were held for 8.86 s +/- 7.06 s or less. The average maximum grasp time during these tasks was 13.37 s +/- 11.42 s. These results form the basis for determining how abdominal tissues are to be mechanically tested in ranges and durations of force and deformation that are surgically realistic. Additionally, this information may serve as design specifications for new surgical robots or haptic simulators. PMID- 15544238 TI - Reconstruction and enhancement in monocular laparoscopic imagery. AB - Constrained minimally-invasive surgical environments create a number of challenges for the surgeon and for automated tools designed to aid in the performance and analysis of complex procedures. The 3D reconstruction of the operative field opens up a number of possibilities for immersive presentation, automated analysis, and post-operative evaluation of surgical procedures. This paper presents a method for estimating complete 3D information about scope and instrument positioning from monocular imagery. These measurements can be used as the basis for deriving and presenting additional cues during procedures, and can also be used for post-procedure analysis such as objective estimates of high level performance measures like economy of motion and ergonomic metrics. PMID- 15544239 TI - Patient specific dynamic geometric models from sequential volumetric time series image data. AB - Generating patient specific dynamic models is complicated by the complexity of the motion intrinsic and extrinsic to the anatomic structures being modeled. Using a physics-based sequentially deforming algorithm, an anatomically accurate dynamic four-dimensional model can be created from a sequence of 3-D volumetric time series data sets. While such algorithms may accurately track the cyclic non linear motion of the heart, they generally fail to accurately track extrinsic structural and non-cyclic motion. To accurately model these motions, we have modified a physics-based deformation algorithm to use a meta-surface defining the temporal and spatial maxima of the anatomic structure as the base reference surface. A mass-spring physics-based deformable model, which can expand or shrink with the local intrinsic motion, is applied to the metasurface, deforming this base reference surface to the volumetric data at each time point. As the meta surface encompasses the temporal maxima of the structure, any extrinsic motion is inherently encoded into the base reference surface and allows the computation of the time point surfaces to be performed in parallel. The resultant 4-D model can be interactively transformed and viewed from different angles, showing the spatial and temporal motion of the anatomic structure. Using texture maps and per vertex coloring, additional data such as physiological and/or biomechanical variables (e.g., mapping electrical activation sequences onto contracting myocardial surfaces) can be associated with the dynamic model, producing a 5-D model. For acquisition systems that may capture only limited time series data (e.g., only images at end-diastole/end-systole or inhalation/exhalation), this algorithm can provide useful interpolated surfaces between the time points. Such models help minimize the number of time points required to usefully depict the motion of anatomic structures for quantitative assessment of regional dynamics. PMID- 15544240 TI - GiPSi: an open source/open architecture software development framework for surgical simulation. AB - In this paper we propose an open source/open architecture framework for developing organ level surgical simulations. Our goal is to facilitate shared development of reusable models, to accommodate heterogeneous models of computation, and to provide a framework for interfacing multiple heterogeneous models. The framework provides an intuitive API for interfacing models with spatial relationships. It is specifically designed to be independent of the specifics of the modeling methods used and therefore facilitates seamless integration of heterogeneous models and processes. Furthermore, each model has separate geometries for visualization, simulation, and interfacing, allowing the modeler choose the most natural geometric representation for each case. PMID- 15544241 TI - Real-time volume haptic rendering of non-linear viscoelastic behavior of soft tissue through dynamic atomic unit approach. AB - The aim of computer haptics is to enable the user to touch, feel and maneuver virtual objects using a haptic interface. As the user "feels" the virtual object by applying force through the interface, complex calculations have to be done in real-time to generate a feedback force appropriate to the material properties of the object being "touched". In this paper we propose a method for modeling soft bodies, which incorporate non-linear, viscoelastic, anisotropic behavior that will enable real-time user interaction and still satisfy the high force-feedback frequency requirements. In this paper, we restrict the user interaction with virtual objects to palpation. PMID- 15544242 TI - Study of soft tissue cutting forces and cutting speeds. AB - A versatile equipment to study the cutting of soft tissue with surgery scalpel was designed and constructed. Experiments were performed with pig liver (ex-vivo) to measure the blade-tissue interaction forces at cutting speeds ranging from 0.1 cm/sec-2.54 cm/sec. The experimentally measured force-displacement curves reveal that the liver cutting process was made up of a sequence of repeating local units with similar features. Each local unit was comprised of a linear deformation phase followed by a crack growth phase. A method was developed to quantify the deformation resistance of the tissue during each local deformation phase in cutting. This deformation resistance was presented in the form of a self consistent local effective Young's modulus (LEYM), and was determined by post processing force-displacement data with finite element models. Values for LEYM were determined from plane-stress finite element model and plane-strain finite element model. The plane-stress LEYM values were within a close bound of the plane-strain values. Results of the self-consistent LEYM at different cutting speeds show that the tissue's resistance to deformation decreased as the cutting speed increased. PMID- 15544243 TI - Level set based auto segmentation of the tagged left ventricle MR images. AB - To facilitate automatic segmentation, we adopt SVM (Support Vector Machine) to localize the left ventricle, and the segmentation is then carried out with narrow band level set. The method of generating the narrow band is improved such that the time used is reduced. Based on the imaging characteristics of the tagged left ventricle MR images, BPV (block-pixel variation) and intensity comparability are introduced to improve the speed term of level set and to increase the precision of segmentation. Our method can perform the segmentation of the tagged left ventricle MR images accurately and automatically. PMID- 15544244 TI - Interactive cutting simulation with adaptive refinements using digital logic design analogy. AB - A major requirement for surgical simulation is to allow virtual tissue cutting. This paper presents a scalable and adaptive cutting technique based on a mass spring mesh. By the analogy of digital logic design, an arbitrary incision is modeled systematically by translating the cutting process into a state diagram. Subdivision of mesh elements is driven by the state transitions. Node redistribution, local re-meshing and deformation are applied to refine the subdivided mesh. PMID- 15544245 TI - A one degree of freedom haptic system to investigate issues in human perception with particular application to probing tissue. AB - This paper presents the results of several early studies relating to human haptic perception sensitivity when probing a virtual object. A 1 degree of freedom (DoF) rotary haptic system, that was designed and built for this purpose, is also presented. The experiments were to assess the maximum forces applied in a minimally invasive surgery (MIS) procedure, quantify the compliance sensitivity threshold when probing virtual tissue and identify the haptic system loop rate necessary for haptic feedback to feel realistic. PMID- 15544246 TI - Workflow modeling and analysis of computer guided prostate brachytherapy under MR imaging control. AB - We demonstrate that classical Business Process Reengineering (BPR) methods can be successfully applied to Computer Aided Surgery while increasing safety and efficiency of the overall procedure through an integrated Workflow Management System. Computer guided Prostate Brachytherapy, as a sophisticated treatment by an interdisciplinary team, is perfectly suited to apply our method. Detailed suggestions for improvement of the whole procedure could be derived by our modified BPR method. PMID- 15544247 TI - The compensation of head motion artifacts using an infrared tracking system and a new algorithm for fMRI. AB - We aim to provide a next generation Magnetic Resonance Imaging (MRI) technology with an integrated solution for reducing motion artifacts in brain imaging applications. New developments in the field of MRI are revolutionizing the diagnostic capabilities e.g. of functional (fMRI) of the technique. Unfortunately, motion artifacts are eminent problems in cerebral MRI images, especially in difficult patient populations (e.g. chronic pain, children, neonates). Patient motion artifacts are present in 2D sequences, but are extremely detrimental in multi-slice 3D sequences often employed in fMRI. The problem of motion compensation in MRI technology deals with: Identification of the source as well as pattern of motion. Obtaining a mathematical model of motion that can be used to identify and then compensate the motion effects. Optimizing the image acquisition sequence in order to minimize, or even eliminate, the effect of motion. We propose a method to obtain a quantitative measure of the movement of the head between different data acquisition points in both MRI, and functional MRI examination. PMID- 15544248 TI - Real-time synchronization of kinematic and video data for the comprehensive assessment of surgical skills. AB - Surgical dexterity in operating theatres has traditionally been assessed subjectively. Electromagnetic (EM) motion tracking systems such as the Imperial College Surgical Assessment Device (ICSAD) have been shown to produce valid and accurate objective measures of surgical skill. To allow for video integration we have modified the data acquisition and built it within the ROVIMAS analysis software. We then used ActiveX 9.0 DirectShow video capturing and the system clock as a time stamp for the synchronized concurrent acquisition of kinematic data and video frames. Interactive video/motion data browsing was implemented to allow the user to concentrate on frames exhibiting certain kinematic properties that could result in operative errors. We exploited video-data synchronization to calculate the camera visual hull by identifying all 3D vertices using the ICSAD electromagnetic sensors. We also concentrated on high velocity peaks as a means of identifying potential erroneous movements to be confirmed by studying the corresponding video frames. The outcome of the study clearly shows that the kinematic data are precisely synchronized with the video frames and that the velocity peaks correspond to large and sudden excursions of the instrument tip. We validated the camera visual hull by both video and geometrical kinematic analysis and we observed that graphs containing fewer sudden velocity peaks are less likely to have erroneous movements. This work presented further developments to the well-established ICSAD dexterity analysis system. Synchronized real-time motion and video acquisition provides a comprehensive assessment solution by combining quantitative motion analysis tools and qualitative targeted video scoring. PMID- 15544249 TI - Visualization of two image variables simultaneously using cardinal directions of color vision. AB - Visualization of medical image information can be achieved by using color scales to enhance aspects of the data. We have used the cardinal directions of color to make a continuous and representation of phase data that wraps around every 360 deg, and added in another dimension using luminance to illustrate amplidute. PMID- 15544250 TI - Soft tissue deformation using a Hierarchical Finite Element Model. AB - Simulating soft tissue deformation in real-time has become increasingly important in order to provide a realistic virtual environment for training surgical skills. Several methods have been proposed with the aim of rendering in real-time the mechanical and physiological behaviour of human organs, one of the most popular being Finite Element Method (FEM). In this paper we present a new approach to the solution of the FEM problem introducing the concept of parent and child mesh within the development of a hierarchical FEM. The online selection of the child mesh is presented with the purpose to adapt the mesh hierarchy in real-time. This permits further refinement of the child mesh increasing the detail of the deformation without slowing down the simulation and giving the possibility of integrating force feedback. The results presented demonstrate the application of our proposed framework using a desktop virtual reality (VR) system that incorporates stereo vision with integrated haptics co-location via a desktop Phantom force feedback device. PMID- 15544251 TI - The personal digital assistant (PDA) as a tool for telementoring endoscopic procedures. AB - The telementoring of surgical procedures is currently achieved via a wired infrastructure that usually requires sophisticated videoconference systems. This project represents the first step in assessing the potential for using handheld computers as a mobile alternative to current telementoring systems. Specifically, this project compares a handheld computer to a standard CRT monitor regarding their capability to accurately display video images from an endoscopic procedure. Video images from two previously recorded endoscopic procedures were transmitted from a standard VCR to: 1) a handheld computer (iPAQ 3670 running Pocket PC) via a wireless LAN and 2) a standard CRT monitor via a wired analog connection. The software-used on the handheld device was custom designed to allow 320 X 240 pixel video images to be broadcast in real time. Twenty-three surgical residents who had completed an endoscopy rotation were randomized to watch one of the two videotaped endoscopic procedures on the hand held computer or on the CRT monitor. After viewing the procedure, a ten-question quiz was used to assess the ability of each participant to recognize several anatomic landmarks. The result of each questionnaire was expressed as the percentage of correct responses. Using a crossover design, each participant then viewed the other videotaped procedure using the alternate device and completed a second quiz. The mean test score for each device was calculated, and these data was analyzed using a Student T test. The observed difference between the mean test score associated with the handheld device (77.93 +/- 11.26) and the CRT monitor (81.30 +/- 12.54) was not statistically significant (p<0.41). In addition, regardless of the device used, scores corresponding to video tape one were significantly higher than those recorded for video tape two (84.35 +/- 9.92 vs. 74.35 +/- 11.61; p < 0.01) All participants were able to recognize anatomic landmarks equally well when viewing broadcasted endoscopic procedures on a handheld display or a standard CRT monitor. Handheld computers may have a role in telementoring residents who are performing endoscopic procedures. Further research is needed to evaluate the integration of handheld devices into telementoring and robotic system to perform surgical procedures. PMID- 15544252 TI - Augmented reality laser projection device for surgery. AB - We have developed an augmented reality system capable of projecting preoperative plans directly onto a patient using rapidly scanned laser beams. Projected contours can typically represent cut paths, tumors or delineate boundaries of interest. The system can be used as part of, or a replacement for, conventional robotic Telesurgery systems. Because the graphics are projected, there is no degradation in surgeon's view due to optical components interposed between the surgeon's eye and the patient. This system has been designed to work with a common infrared 3D camera system used in image-guided surgery, and projects both visible and infrared beams. The IR beam enables surface digitization functions to be carried out using the camera. The clinical accuracy is in the range required by CAS procedures, around 1-2mm. The device will be particularly useful for executing precise preoperative plans and for teleconsultation applications, where planned or live consultations can be efficiently communicated to a less skilled local caregiver. PMID- 15544253 TI - StapSim: a virtual reality-based stapling simulator for laparoscopic hemiorrhaphy. AB - The growing interest in laparoscopic hernia surgery and in surgical simulators has motivated our current research. In this paper, we present our work in simulating the process of stapling used in laparoscopic herniorrhaphy. By connecting two separate deformable preperitoneal meshes together, our model has simulated the repair process for a bilateral hernia correction. The task of mesh placement and stapling of corners was simulated, to allow surgeons to practice their hand-eye coordination. Various deformable models and numerical methods were researched to comply with the real time requirements. The stapling simulator can either be independently used as a part-task (sub-task) trainer or be integrated as a module of a complete VR-based simulator. A phantom device was used to provide haptic-based force feedback during task rehearsals. PMID- 15544254 TI - Using cognitive task analysis to facilitate collaboration in development of simulator to accelerate surgical training. PMID- 15544255 TI - Surgical training using haptics over long internet distances. AB - This paper describes a collaborative surgical training prototype using haptics, which has been able to operate across the world. It allows two users to collaboratively manipulate a simulation of pliable human body organs, as well as guide each other's 'hands' over 22,000 km of internet connection. It uses a force impulse collection mechanism feeding haptics data to a single physics server program. The server runs a 'pseudo' physics model that is resilient to latency. PMID- 15544256 TI - Realistic anatomical texture for laparoscopic surgery simulation. AB - Visual realism in laparoscopic surgery simulation is very desirable. Previously, much work has been done to extract the organs and textures from the visible human data (VHD) using various rendering techniques. We present here a technique to extract the true texture from visible human data for the laparoscopic herniorrhaphy simulation. A VHD slice provides texture only in the direction in which it was visualized whereas the surgeons visualize the facet of an organ during a laparoscopic surgery. Our paper describes an approach for the extraction and mapping facet texture from the VHD. PMID- 15544257 TI - Challenges realising effective radiological interventional virtual environments: the CRaIVE approach. PMID- 15544258 TI - Virtual reality based system for training on knee arthroscopic surgery. AB - Surgical training systems based on virtual reality (VR) and simulation techniques offer a cost-effective and efficient alternative to traditional training methods. This paper describes a virtual reality system for training arthroscopic knee surgery. The virtual model used in this system is constructed from the Visual Human Project dataset. The system simulates the real-time deformation of soft tissue with topological change using finite element analysis. To offer the realistic tactile feedback, we construct a specialized force feedback hardware. PMID- 15544259 TI - Visualization of muscle function for medical education. AB - We developed a muscle shape model with a view to visualizing muscle functionality for medical education purposes. In our model a muscle is represented by a set of fibers, which run the length of the body of the muscle from origin to insertion. The fiber curves are equipped with contractile and elastic elements and are used as deformation guides that re-shape the muscle surface. This enables us to specify the timing and the level of contraction of a muscle as well as its volume and shape changes. The model can be configured to animate muscles, tendons and ligaments. EMG (electromyography) data have been used to drive the contractile element of the model. We researched and implemented 34 basic skeleton movements and 4 gross motor movements involving 15 joints and over 165 muscles in the human body. Complex bone movements were based on motion captured data combined with live video reference. We used interactive movies in order to present a bone/muscle action from a wide range of views. PMID- 15544260 TI - Automated kinematic generator for surgical robotic systems. AB - Unlike traditional assembly line robotic systems that have a fixed kinematic structure associated with a single tool for a structured task, next-generation robotic surgical assist systems will be required to use an array of end-effector tools. Once a robot is connected with a tool, the kinematic equations of motion are altered. Given the need to accommodate evolving surgical challenges and to alleviate the restrictions imposed by the confined minimally invasive environment, new surgical tools may resemble small flexible snakes rather than rigid, cable driven instruments. Connecting to these developing articulated tools will significantly alter the overall kinematic structure of a robotic system. In this paper we present a technique for real-time automated generation and evaluation of manipulator kinematic equations that exhibits the combined advantages of existing methods-speed and flexibility to kinematic change--without their disadvantages. PMID- 15544261 TI - Palpation imaging using a haptic system for virtual reality applications in medicine. AB - In the field of medical diagnosis, there is a strong need to determine mechanical properties of biological tissue, which are of histological and pathological relevance. Malignant tumors are significantly stiffer than surrounding healthy tissue. One of the established diagnosis procedures is the palpation of body organs and tissue. Palpation is used to measure swelling, detect bone fracture, find and measure pulse, or to locate changes in the pathological state of tissue and organs. Current medical practice routinely uses sophisticated diagnostic tests through magnetic resonance imaging (MRI), computed tomography (CT) and ultrasound (US) imaging. However, they cannot provide direct measure of tissue elasticity. Last year we presented the concept of the first haptic sensor actuator system to visualize and reconstruct mechanical properties of tissue using ultrasonic elastography and a haptic display with electrorheological fluids. We developed a real time strain imaging system for tumor diagnosis. It allows biopsies simultaneously to conventional ultrasound B-Mode and strain imaging investigations. We deduce the relative mechanical properties by using finite element simulations and numerical solution models solving the inverse problem. Various modifications on the haptic sensor actuator system have been investigated. This haptic system has the potential of inducing real time substantial forces, using a compact lightweight mechanism which can be applied to numerous areas including intraoperative navigation, telemedicine, teaching and telecommunication. PMID- 15544262 TI - Virtual patient: a photo-real virtual human for VR-based therapy. AB - A high fidelity Virtual Human Interface (VHI) system was developed using low-cost and portable computers. The system features real-time photo-realistic digital replicas of multiple individuals capable of talking, acting and showing emotions and over 60 different facial expressions. These virtual patients appear in a high performance virtual reality environment featuring full panoramic backgrounds, animated 3D objects, behavior and AI models, a complete vision system for supporting interaction and advanced animation interfaces. The VHI takes advantage of the latest advances in computer graphics. As such, it allows medical researchers and practitioners to create real-time responsive virtual humans for their experiments using computer systems priced under $2000. PMID- 15544263 TI - Effect of sensory substitution on suture manipulation forces for surgical teleoperation. AB - Bilateral telemanipulation, which applies haptic feedback to the operator, is not yet available in most commercial robot-assisted surgical systems. We have shown in previous work that the lack of haptic (force or tactile) feedback is detrimental in applications requiring fine suture manipulation. In this paper, we study the effect of substituting direct haptic feedback with visual and auditory cues. Using the da Vinci robot from Intuitive Surgical, we observed the difference between applied forces during a knot tying procedure for four different sensory feedback substitution scenarios: no feedback, auditory feedback, visual feedback, and a combination of auditory and visual feedback. Our results indicate that visual feedback, which provides continuous force information, would improve robot-assisted performance during complex surgical tasks such as knot tying with fine sutures. Discrete auditory feedback gives additional useful support to the surgeon. PMID- 15544264 TI - Chemical agent simulator for emergency preparedness training. PMID- 15544265 TI - A study of overlapping virtual objects on real objects in 3D space. AB - Augmented reality (AR) is anticipated for use in medical practice. However, virtual objects cannot be overlaid exactly on real objects using theoretical data. Therefore, this study is intended to determine the correction method to overlay a virtual object with real object accurately. Experimental results confirmed that differences of perceived depth position between real and virtual objects can be made negligible (within 1 mm on average) by correcting sizes of virtual objects such that they can be seen identically to the real object. PMID- 15544266 TI - Optimization of a vector quantization codebook for objective evaluation of surgical skill. AB - Surgical robotic systems and virtual reality simulators have introduced an unprecedented precision of measurement for both tool-tissue and tool-surgeon interaction; thus holding promise for more objective analyses of surgical skill. Integrative or averaged metrics such as path length, time-to-task, success/failure percentages, etc., have often been employed towards this end but these fail to address the processes associated with a surgical task as a dynamic phenomena. Stochastic tools such as Markov modeling using a 'white-box' approach have proven amenable to this type of analysis. While such an approach reveals the internal structure of the of the surgical task as a process, it requires a task decomposition based on expert knowledge, which may result in a relatively large/complex model. In this work, a 'black box' approach is developed with generalized cross-procedural applications., the model is characterized by a compact topology, abstract state definitions, and optimized codebook size. Data sets of isolated tasks were extracted from the Blue DRAGON database consisting of 30 surgical subjects stratified into six training levels. Vector quantization (VQ) was employed on the entire database, thus synthesizing a lexicon of discrete, task-independent surgical tool/tissue interactions. VQ has successfully established a dictionary of 63 surgical code words and displayed non-temporal skill discrimination. VQ allows for a more cross-procedural analysis without relying on a thorough study of the procedure, links the results of the black-box approach to observable phenomena, and reduces the computational cost of the analysis by discretizing a complex, continuous data space. PMID- 15544267 TI - The development of a VR system for the cognitive & behavioral assessment of schizophrenia. AB - Patients with schizophrenia have many trouble in their life due to their thinking disorders such as delusion or hallucination. It is because they have a deficit in the ability which to systematize and integrate information. Therefore, they cannot integrate or systematize properly the integrated stimuli occurring frequently in a real situation. In this study, we suggest a virtual reality system for the assessment of their ability to integrate, memorize and react to integrated stimuli. The virtual reality system we developed provided multimodal stimuli, such as visual and auditory stimuli or visuo-spatial integration, and the patients was assessed and compared their thought flexibility and working memory abilities based on multi-modal manner by making the patient experience and react to multimodal stimuli. From this study, we could conclude that the VR could provide a patient with various stimuli in an immersive environment and allow the assessment of cognitive ability, and the identification of the relationships between cognitive functions. PMID- 15544268 TI - FEM-based interaction model between elastic objects for indirect palpation simulator. AB - Indirect palpation is required to examine lots of cases like breast cancer and prostate malignance. This paper proposes interaction model between elastic objects to simulate indirect palpation. The interaction is simulated by displacement of colliding elements based on normal stress derived from temporary displacement. The physics-based approach represents the difference of physical properties such as stiffness of colliding objects. Deformation and haptic reproduction is possible to be carried out in real time with two organ models consisting of roughly 200 nodal points. As an example, we developed a rectal palpation simulator based on the proposed method. The experiment using rectal palpation simulator confirmed that the method enables a user to perceive difference of stiffness of prostate model located behind rectum model indirectly. PMID- 15544269 TI - Visualization of the variability of 3D statistical shape models by animation. AB - Models of the 3D shape of anatomical objects and the knowledge about their statistical variability are of great benefit in many computer assisted medical applications like images analysis, therapy or surgery planning. Statistical model of shapes have successfully been applied to automate the task of image segmentation. The generation of 3D statistical shape models requires the identification of corresponding points on two shapes. This remains a difficult problem, especially for shapes of complicated topology. In order to interpret and validate variations encoded in a statistical shape model, visual inspection is of great importance. This work describes the generation and interpretation of statistical shape models of the liver and the pelvic bone. PMID- 15544270 TI - Introducing a novel haptic interface for the planning and simulation of open surgery. AB - Existing simulation software, originally developed for the simulation and planning of inguinal hernia repair, was fused with two haptic feedback devices: the SensAble Technologies Phantom Desktop, and the ACROE/ICA Telluris system. The former allows easy integration on a PC-based platform, though in the long run, the Telluris system is preferred because of its robustness and its ability to design custom-built solutions. PMID- 15544271 TI - A new form of application service provider (ASP) in the development of data systems and data flow and management of data in (multiple campus) health care institutions. AB - There is a distinct lag in the way the informatics management is applied, implemented and used in health care institutions in comparison to other industries. Part of this is caused by strict regulations, but there also seems to be a defined trust and attitude problem from the medical corps towards dependency from automatisation. The only way to guarantee overall advance however is to apply technology implementations which enhance the performance capabilities of the medical corps, the way they already have in other fields. We propose a new model for implementing software applications and their communication on the scale of a multi campus health care institution. PMID- 15544272 TI - Argonaute 3D: a real-time cooperative medical planning software on DSL network. AB - Today, diagnosis of cancer and also therapeutic choice imply many specialized practitioners. They are generally located at different places and have to take the best decision as promptly as possible with the difficulty of CT-scan or MRI interpretation. Argonaute 3D is a tool that easily overcomes these issues, thanks to a cooperative solution based on virtual reality. An experimentation, where four practitioners met virtually throughout France, allowed to assess the interest of this solution. PMID- 15544273 TI - Realistic simulation of surgical cutting of soft tissues in real time with force feedback. AB - In this paper we present a geometrically efficient algorithm coupled with a physically based numerical scheme using a meshfree approach for the simulation of surgical procedures including surgical cutting in multimodal virtual environments. Progressive cutting, without the generation of new primitives, is achieved by snapping the nearest nodes to the interaction point between the cutting tool and the underlying polygon edge. The realism of the simulations is enhanced by employing a local subdivision algorithm in the vicinity of the tool tissue interaction region and a cutting gutter to display the interior structure of the soft tissue as the cut opens up. A meshfree method is used to compute the deformation fields and interaction forces. PMID- 15544274 TI - Two-handed next generation suturing simulator. AB - A realistic two-handed surgical suturing simulation would have a broad impact on the way doctors are trained in the field of surgery. Current state-of-the-art in suturing simulators do not accurately represent realistic suturing conditions, leaving many necessary components, i.e. two-handed interaction, skin undermining, skin repositioning etc., out of the model completely. The goal of this research is to develop a next generation immersive suturing simulator that uses an integrated two-handed tension based haptic device for tactile feedback, a hi fidelity finite element model for more accurate skin, tissue and needle modeling and stereoscopic vision for depth-of-field. PMID- 15544275 TI - Bioterrorism: development of large-scale medical readiness using multipoint distance-based simulation training. AB - Accordingly to HIRSA, 35,000 health professionals need to be trained in recognition and acute field treatment of victims of bioterrorism within year 2004 alone The Department of Defense anticipates even larger numbers. Training of very large number of healthcare workers is particularly daunting in the context of "just-in-time" education. The paper presents utilization of simulation-based distance training as a particularly useful tool in rapid development of readiness in a large population of widely distributed medical and lay personnel facing imminent threat of a chem/bioterrorism incident. PMID- 15544276 TI - Training and assessment of psychomotor skills for performing laparoscopic surgery using BEST-IRIS virtual reality training simulator. AB - In this article, we present the results of a pilot study that examined the performance of people training on a Virtual Reality based BEST-IRIS Laparoscopic Surgery Training Simulator. The performance of experienced surgeons was examined and compared to the performance of residents. The purpose of this study is to validate the BEST-IRIS training simulator. It appeared to be a useful training and assessment tool. PMID- 15544277 TI - A one-DOF freehand haptic device for robotic tele-echography. AB - Tele-echography may be an interesting alternative to conventional care when the medical expert is not close to the patient to be examined. In a system for tele echography, medical expert proprioception and gesture feelings must be preserved to synchronize the ultrasound images with the motion made by the physician. It is why we develop a master-slave system for robotic tele-echography integrating force feedback. Because existing haptic devices are not fully adapted to this application--their workspace is often small and their basis is fixed--, we have developed a one-dimensional free-hand force feedback device. In this paper, we describe the design and a first evaluation of this new haptic device for robotic tele-echography. PMID- 15544278 TI - In-vivo stereoscopic imaging system with 5 degrees-of-freedom for minimal access surgery. AB - Endoscopic imaging for minimal access surgery has many limitations that include: 2D and narrow angle imaging, limited workspace of the endoscope caused by the fulcrum effect of the body wall, and the presence of the endoscope in the incision that prevents use of the incision for other instrumentation. We have designed a novel stereoscopic 3D imaging device with 5 DOF and remote control that can be inserted and attached in the body cavity. The device, contained within a 11/16" tube, includes two miniature cameras and five small motors that position the cameras to provide a stereoscopic view of the surgical site. When inserted the cameras are retracted and protected by an outer shell. After the device is fixed within the abdominal cavity, a motor rotates an inner shell to expose the cameras. Once exposed, the cameras can tilt in tandem, translate independently along the axis of the tube, and independently pan. The software controls the cameras to create new views for the surgeon, to move along the adjustable baseline, to verge for stereoscopic viewing, and to potentially track moving organs. We have completed a proof of concept design, which includes CAD models and animations of the device, and we are currently building a physical prototype. Once the prototype is completed, we will begin testing it in a surgical mock-up, followed by animal and clinical trials. PMID- 15544279 TI - Ensuring the usability of a knee arthroscopy simulator. AB - There is limited research considering the usability of medical virtual environments. Usability evaluation is an essential validation phase that considers the extent to which a product achieves its specific goals, with effectiveness, efficiency and satisfaction. A four-stage iterative approach is adopted to enhance usability in the development of a knee arthroscopy training system. This process has drawn attention to issues that may impede system usability for example non-conformity to platform conventions, and visibility of the system status. The process highlights features that computer scientists can overlook when working closely with a system but that are essential to user acceptance and effective application. PMID- 15544280 TI - A flexible virtual reality tutorial for the training and assessment of arthroscopic skills. AB - Through definition of a comprehensive tutorial model, the Warwick, Imperial and Sheffield Haptic Knee Arthroscopy Training System (WISHKATS) aims to provide independent, flexible and consistent training and assessment. The intention is to satisfy user acceptance by limiting the constraints by which the system can be utilised, as well as demonstrating validity and reliability. System use can either be under the guidance and feedback offered by the system or of a senior surgeon. Objective metrics are defined for performance feedback and formal assessment. PMID- 15544281 TI - Evaluation of the benefit of VR simulation in a multi-media web-based educational tool. AB - WebSET is an Internet based educational tool that can be used on any standard personal computer. It has been developed by a European collaboration and integrates high quality multimedia learning materials with VR simulation. The aim of this study was to evaluate the benefit of the VR simulation on the learning of procedure based psychomotor skills. Subjects were divided into three groups. The group that used the entire package including the VR simulation were superior to the group that used only the multimedia component in terms of the their procedural skills in the post-training assessment. Both groups performed better than the controls. PMID- 15544282 TI - Parameter optimisation for the behaviour of elastic models over time. AB - Optimisation of parameters for elastic models is essential for comparison or finding equivalent behaviour of elastic models when parameters cannot simply be transferred or converted. This is the case with a large range of commonly used elastic models. In this paper we present a general method that will optimise parameters based on the behaviour of the elastic models over time. PMID- 15544283 TI - LR-Spring Mass model for cardiac surgical simulation. AB - The purpose of the research conducted was to develop a real-time surgical simulator for preoperative planning of surgery in congenital heart disease. The main problem simulating procedures on cardiac morphology is the need for a large degree of detail and simulation speed. In combination with a demand for physically realistic real-time behaviour this gives us tradeoffs not easily balanced. The LR-Spring Mass model handles these constraints by the use of domain specific knowledge. PMID- 15544284 TI - Distributed interactive virtual environments for collaborative medical education and training: design and characterization. AB - Project TOUCH (Telehealth Outreach for Unified Community Health) is a collaborative effort between University of New Mexico and University of Hawaii. The purpose of the project is to demonstrate the feasibility of using advanced technologies to overcome geographical barriers to delivery of medical education and to enhance the learning process within a group setting. This has led to the design and implementation of a new system that addresses the critical requirements for collaborative virtual environments: consistency, networking, scalability, and system integration. The objective of this study is to evaluate the performance of the collaborative system based on use patterns during Project TOUCH sessions. PMID- 15544285 TI - Physics-based preoperative approach planning using hybrid virtual bodies. AB - This paper proposes a hybrid model mixing geometry and volume data to improve representation of virtual bodies. This model applies object-oriented data models and rendering techniques to virtual organs, and enables both interactive VR simulation and detailed volume visualization of tissue of interest (e.g. coronary). Also, a physics-based framework interactively simulates estimated surgical fields which are used in preoperative discussion. Based on the proposed methods, a VR-based strategic planning system is developed. The system does not need high cost manual segmentation of patient dataset and efficiently supports planning of surgical approaches in cardiovascular surgery. PMID- 15544286 TI - Practical haptic navigation with clickable 3D region input interface for supporting master-slave type robotic surgery. AB - Conventional display in robotic surgery such as flat displays or stereoscopic displays decreases obtainable information around target tissue. For supporting manipulation and performing safe surgery, this paper proposes a haptic navigation method, which enables surgeons to avoid collision with untouchable regions around target tissue by producing force feedback through a master manipulator. This paper also developed an input interface for assignment of 3D untouchable regions through 2D device. Simulator based experiment clears effectiveness of the proposed haptic navigation for improving safety of robotic surgery. PMID- 15544287 TI - THUMP: an immersive haptic console for surgical simulation and training. AB - Telerobotic systems are revolutionizing minimally invasive surgery (MIS), giving the surgeon complete control over precise dexterous movements of tiny robotic instruments. Such 'surgery-by-wire' approaches also create unique opportunities for simulation and training, as the surgeon operates at a computer-mediated haptic console. Possible extensions include offline training in simulated environments and advanced guidance and mentoring during actual operations. To explore these options and further improve telerobotic interfaces, we have constructed a two-handed, fully articulating haptic console that provides force and torque feedback as well as a stereoscopic display. PMID- 15544288 TI - Ergonomic interface concepts for minimally invasive, remote, and virtual surgical systems. AB - Traditional open surgical techniques require a surgeon to assume a posture of leaning over the patient with a direct eye-to-hand perspective. As new minimally invasive and remote surgical procedures evolve, the surgeon is not required to maintain the same posture as in open techniques. While more ergonomic postures may be facilitated, some current remote systems have maintained surgeon configurations that are small variants of legacy postures (e.g., maintaining the eye to hand perspective). While the legacy configuration may be more familiar with some surgeons, studies have indicated that it can result in excessive fatigue. Robotics and human factors researchers have determined that fatigue due to inefficiencies in operator interfaces lead to longer completion times and increased task execution errors. This paper discusses operator interface design issues and guidelines that are relevant to remote and minimally invasive surgery, and presents one possible operator interface solution based on the compact remote console deployed for environmental restoration and remote handling of hazardous nuclear waste. PMID- 15544289 TI - Virtual reality combats test anxiety: a case study report. AB - This pilot study is the first known in-depth case study of the effectiveness of virtual reality therapy (VRT) as a treatment for Test Anxiety (TA). The subject of the study was a 28-year-old male, whose anxiety and avoidance behavior was interfering with his normal academic activities. For treatment, he was placed in a virtual classroom and later in a virtual auditorium. The subject was exposed to six moderately increasing in difficulty level virtual situations. The subject rated each situation for discomfort. As a simple measure of anxiety, a modified version of the Subjective Units of Disturbance (SUD) scale was used every five minutes during exposure. This case study showed VRT to be an effective treatment method for reducing self-reported TA. Symptoms experienced by the subject during VRT sessions were just as real to the subject as actual test taking and general TA situations. They included increased heart rate, mild dizziness, and headaches. This case study of TA indicates that VRT may be used as an effective treatment method for reducing self-reported anxiety and improving the performance of subject(s) who suffer from TA. PMID- 15544290 TI - The delivery simulator: a new application of medical VR. AB - This paper presents an elementary overview of the potential of Multimodal Virtual Reality (MVR) techniques in medical education, e.g. obstetrics. The study shows how to transfer the concept of MVR from a time-independent environment, e.g. the Munich Knee Joint Simulator, to a time-critical simulation environment as it can be found in flight simulators. The simulator consists of a haptic, a graphical and an acoustic user interface, which are connected to a biomechanical model for the birth process itself and a physiological model of both mother and child, in order to simulate, e.g. a cardiotocograph (CTG). The user can just watch an uncomplicated birth or is acting as the responsible obstetrician who has a variety of treatment options during the delivery with the most relevant medication or forceps/vacuum-extraction. During this practical training a MVR feedback system assists the trainee and exposes his errors and, thus, allows him to learn faster without endangering a real mother and her child. This concept allows for the first time to transfer stored haptic expert-knowledge to the trainee without a tool-based feedback approach. PMID- 15544291 TI - Development of a novel surgical support instrument and virtual system incorporating new tactile sensor technology. AB - Surgical practice would be significantly enhanced with robotic systems incorporating tactile sensors. Current tactile sensor technology consists mainly of strain gauge elements having a limited bandwidth. A novel tactile sensor system, has been developed using a piezoelectric transducer(PZT), to simulate the properties of the human hand for use as a surgical support instrument and a palpation probe. Visualization of tactile information as an audio signal is provided, representing tissue properties in terms of an amplitude and frequency modulated signal. Representative data measured from pig brain, lung, pancreas, tongue and liver show that the changes in frequency corresponds to tissues stiffness and contact pressure. The technology developed in this new surgical support system has potential applications in virtual systems or robotic tele medical care. PMID- 15544292 TI - Development and human factors analysis of neuronavigation vs. augmented reality. AB - This paper is focused on the human factors analysis comparing a standard neuronavigation system with an augmented reality system. We use a passive articulated arm (Microscribe, Immersion technology) to track a calibrated end effector mounted video camera. In real time, we superimpose the live video view with the synchronized graphical view of CT-derived segmented object(s) of interest within a phantom skull. Using the same robotic arm, we have developed a neuronavigation system able to show the end-effector of the arm on orthogonal CT scans. Both the AR and the neuronavigation systems have been shown to be within 3mm of accuracy. A human factors study was conducted in which subjects were asked to draw craniotomies and answer questions to gage their understanding of the phantom objects. The human factors study included 21 subjects and indicated that the subjects performed faster, with more accuracy and less errors using the Augmented Reality interface. PMID- 15544293 TI - Passive vs. active virtual reality learning: the effects on short- and long-term memory of anatomical structures. AB - This pilot study compares the differences in learning outcomes when students are presented with either an active (student-centered) or passive (teacher-centered) virtual reality-based anatomy lesson. The "active" lesson used UCSD's Anatomic VisualizeR and enabled students to interact with 3D models and control presentation of learning materials. The "passive" lesson used a digital recording of an anatomical expert's tour of the same VR lesson played back as a QuickTime movie. Subsequent examination of the recall and retention of the studied anatomic objects were comparable in both groups. Issues underlying these results are discussed. PMID- 15544294 TI - BIRD: Bio-Image Referral Database. Design and implementation of a new web based and patient multimedia data focused system for effective medical diagnosis and therapy. AB - This paper presents a low cost software platform prototype supporting health care personnel in retrieving patient referral multimedia data. These information are centralized in a server machine and structured by using a flexible eXtensible Markup Language (XML) Bio-Image Referral Database (BIRD). Data are distributed on demand to requesting client in an Intranet network and transformed via eXtensible Stylesheet Language (XSL) to be visualized in an uniform way on market browsers. The core server operation software has been developed in PHP Hypertext Preprocessor scripting language, which is very versatile and useful for crafting a dynamic Web environment. PMID- 15544295 TI - The Surgical Illustrator: a web enabled computer program for documenting clinical and procedural details. AB - The medical record not only stores information on actions taken regarding patient care but it is also a source of education for those who read it. Nurses, residents, interns, students and consulting clinicians look to the medical record to gain an understanding of clinical disease and the diagnostic studies and treatment regimens used to affect the disease. We have presented our initial findings and our framework for developing and evaluating The Surgical Illustrator, a software program that will enable clinicians to include in EMRs information that is usually hand drawn in traditional medical records. The future of the EMR will be a direct result of research and development devoted to creating innovative means of conveying clinically pertinent data. Our goal is to make a major contribution to this effort. PMID- 15544296 TI - A virtual reality based navigation system for endovascular surgery. AB - Endovascular surgery provides a minimally invasive solution for the treatment of aortic aneurysms. Fluoroscopic guidance involves X-rays exposure and loss of space information. We have developed a navigation system allowing real-time visualisation of the endovascular tools in a 3D model of the vessels without any radiation exposure. A modified endoprosthesis is equipped with a magnetic sensor tracked by the Aurora magnetic localizer. The registration step uses 2.5D ultrasonography to replace pre-operative CT data in the Operating Room referential. The Virtual Reality based navigation system shows the location of the endoprosthesis inside a 3D CT model of the aorta. Endovascular procedure benefits from a reduced radiation exposure. PMID- 15544297 TI - Catheter simulation system CathI: from patient data generation to cardiological training systems. AB - In this paper we discuss a new approach to generate 3D models for a simulation system for training an angioplasty. The underlying data for these models are obtained from angiograms that are captured during routine interventions in cardiology. For the extraction of the arteries we use a non-linear classificatory with features based on vesselness information (using a scale-space approach), the gray value, and motion information of the arteries. As result we can correctly find 80% of the arteries in the image and we have 4% pixels incorrectly classified as arteries. These models serve for a virtual catheter laboratory that is based on original instruments like catheters, wires, control instruments for the X-ray, syringes, and pressure pumps for the balloon catheter but instead of a patient an input instrument is used. This instrument sends positional and pressure data to a PC that simulates the patient. The cardiologist then obtains the visual and haptic feedback as if we operated a real patient. PMID- 15544298 TI - In vivo robots for laparoscopic surgery. AB - Laparoscopic techniques have allowed surgeons to perform operations through small incisions. However, the benefits of laparoscopy are still limited to less complex procedures because of losses in imaging and dexterity compared to conventional surgery. This project is developing miniature robots to be placed within the abdominal cavity to assist the surgeon. These remotely controlled in vivo robots provide the surgeon with an enhanced field of view from arbitrary angles as well as provide dexterous manipulators not constrained by small incisions in the abdominal wall. PMID- 15544299 TI - A sensorized human torso phantom. AB - Force-torque measuring input devices can significantly enhance the performance of classical simulation environments that are, for example, based on pure passive phantoms. Such devices allow not only the determination of force/torque amplitude and direction but also the contact point on the phantom. The force/torque information can be displayed visually or acoustically, drive a realistic graphical animation environment or it can be saved and compared with a haptic library comprising the force/torque history of any medical specialist. In this paper the technical principle is exemplified by an interactive human torso. A plastic phantom model of a human torso is instrumented with a 6-degree-of-freedom force/torque sensor, thus, allowing an intuitive and interactive use for education of human anatomy. PMID- 15544300 TI - Phantom-based interactive simulation system for dental treatment training. AB - In this paper, we propose a new interactive simulation system for dental treatment training. The system comprises a virtual reality environment and a force-torque measuring device to enhance the capabilities of a passive phantom of tooth anatomy in dental treatment training processes. The measuring device is connected to the phantom, and provides essential input data for generating the graphic animations of physical behaviors such as drilling and bleeding. The animation methods of those physical behaviors are also presented. This system is not only able to enhance interactivity and accessibility of the training system compared to conventional methods but it also provides possibilities of recording, evaluating, and verifying the training results. PMID- 15544301 TI - An adaptive driver and real-time deformation algorithm for visualization of high density lung models. AB - Technological advances in Augmented Reality (AR) and extraction of 3D patient specific medical data led to the creation of medical visualization using AR environments, in which the 3D data is registered and synchronized with the position of the patient. One of the challenges in such visualization environments is maintaining an accurate shape of the 3D data for self-deformable models such as lungs. An accurate deformation of lung model with 3D visualization may significantly increase the teaching and diagnosing ability of physicians. Modeling the deformation of lungs primarily involves the accurate representation of Pressure-volume relationship and the hysteresis in the relationship during inhalation and exhalation. This paper explains a real-time physiologically accurate deformation algorithm and its hardware rendering. We then introduce a novel approach for the representation of accurate pressure volume relationship based on an analogy with classical mechanics. Our simulation results show that the hysteresis obtained is more accurate as compared to current lung models. Thus in our approach a physically realistic deformation of lung model is obtained by the integration of the accurate PV relationship with real-time deformation method. PMID- 15544302 TI - A comparison of the CathSim system and simulated limbs for teaching intravenous cannulation. AB - The present study describes a comparison between the CathSim VR simulator and simulated limbs for training IV cannulation. Two groups of physician assistant students underwent 2 hours of training on either method. Performance was assessed before and after training with a standardized assessment form. The results showed that all students improved after training, but the degree of improvement was greater for those trained with the simulated limbs. These findings may be due to differences between the two training methods as well as the methodology adopted in the present study. PMID- 15544303 TI - Enhancing radiological volumes with symbolic anatomy using image fusion and collaborative virtual reality. AB - Radiological volumes are typically reviewed by surgeons using cross-sections and iso-surface reconstructions. Applications that combine collaborative stereo volume visualization with symbolic anatomic information and data fusions would expand surgeons' capabilities in interpretation of data and in planning treatment. Such an application has not been seen clinically. We are developing methods to systematically combine symbolic anatomy (term hierarchies and iso surface atlases) with patient data using data fusion. We describe our progress toward integrating these methods into our collaborative virtual reality application. The fully combined application will be a feature-rich stereo collaborative volume visualization environment for use by surgeons in which DICOM datasets will self-report underlying anatomy with visual feedback. Using hierarchical navigation of SNOMED-CT anatomic terms integrated with our existing Tele-immersive DICOM-based volumetric rendering application, we will display polygonal representations of anatomic systems on the fly from menus that query a database. The methods and tools involved in this application development are SNOMED-CT, DICOM, VISIBLE HUMAN, volumetric fusion and C++ on a Tele-immersive platform. This application will allow us to identify structures and display polygonal representations from atlas data overlaid with the volume rendering. First, atlas data is automatically translated, rotated, and scaled to the patient data during loading using a public domain volumetric fusion algorithm. This generates a modified symbolic representation of the underlying canonical anatomy. Then, through the use of collision detection or intersection testing of various transparent polygonal representations, the polygonal structures are highlighted into the volumetric representation while the SNOMED names are displayed. Thus, structural names and polygonal models are associated with the visualized DICOM data. This novel juxtaposition of information promises to expand surgeons' abilities to interpret images and plan treatment. PMID- 15544304 TI - Virtual patients in clinical medicine. AB - Virtual Patients are an essential component of medical virtual reality. Most work to date has been dedicated to virtual patients in surgery. However, virtual patients also have a strong potential in clinical medicine, where they can be used as visual interfaces to knowledge-based systems in various simulation and training applications. In this paper, we describe the implementation of a virtual patient in the field of cardiac emergencies and propose directions to extend the development of virtual patients towards the integration of multiple physiological sub-systems. PMID- 15544305 TI - Segmentation and generation of patient-specific 3D models of anatomy for surgical simulation. AB - Patient-specific simulation requires detailed 3D anatomical models. This paper describes how using optimised MRI sequences and clear segmentation criteria, together with adequate segmentation tools can produce high fidelity models able to reflect anatomical variability and allow realistic simulation of operative techniques and conditions. PMID- 15544306 TI - A simple virtual instrument to monitor surgeons' workload while they perform minimally invasive surgery tasks. AB - Monitoring the workload of surgeons while they perform minimally invasive surgery (MIS) tasks can help them learn to reduce effort as they improve performance and can help develop better human-technology interfaces for MIS. To monitor workload, we developed a personal computer based virtual instrument (VI) that uses orientation sensors worn on the surgeon's left and right upper arms to measure upper arm flexion, abduction, and outward rotation angles. From these sensors, we compute indices of effort and integrated effort. One effort index is the upper arm elevation angle. The time integral of this index provides a corresponding integrated effort index. A second effort index is hand velocity. Hand trajectory length is the corresponding integrated effort index. We used the workload monitor VI to study 29 volunteer surgeon subjects while they performed a knot-tying task in a laparoscopic trainer at a standard MIS station. For five of these subjects, we also monitored the workload indices while they performed simulated MIS tasks on a virtual reality Procedicus MIST System. For the subject group, integrated effort, but not level of effort, decreased with increased performance. At each performance level, some subjects worked much harder than others, suggesting that these subjects could benefit by learning to reduce their effort levels. The workload measures from the arm sensors augmented the performance measures provided by the MIST system. PMID- 15544307 TI - Occam's approach to video critical behavior detection: a practical real time video in-vehicle alertness monitor. AB - Driver and pilot fatigue and incapacitation are major causes of injuries and equipment loss. A method is proposed for constant in-vehicle monitoring of alertness, including detection of drowsiness and incapacitation. Novel features of this method include increases in efficiency and specificity that allow real time monitoring in the functional environment by practicable and affordable hardware. The described approach should result in a generally deployable system with acceptable sensitivity and specificity and with capability for operator alarms and automated vehicle intervention to prevent injuries caused by reduced levels of operator performance. PMID- 15544308 TI - A prototype virtual reality system for preoperative planning of neuro endovascular interventions. AB - This paper describes our efforts at creating a system with real time acquisition and geometric processing of the patients vascular anatomy, supplemented by a framework to simulate catheter vasculature interaction. Our system uses biplane angiograms coupled with an algorithm for generation of 3D vascular trees for patient data acquisition. Surface reconstruction and processing of this data is performed to provide novel decision aids to the interventionalist. The processed data can be loaded in a virtual environment to simulate guidance of the catheter through the patient's vasculature. With our combination of techniques, a patient specific visualization and simulation environment can be prepared within the narrow time window available for this procedure. PMID- 15544309 TI - Validation of soft tissue properties in surgical simulation with haptic feedback. AB - Numerous experiments are being conducted to extract soft tissue values for their integration into VR surgical simulations with haptic feedback. Haptic feedback has been shown to be relevant in laparoscopic surgery, however to date no experiments have been conducted to test user sensitivity to changes in soft tissue values in surgical simulations, and how users perception of 'reality' differs from experimentally determined soft tissue values. In this study we conduct a series of experiments investigating haptic sensitivity, haptic differentiation and comparing experimental and empirical values. PMID- 15544310 TI - Comparison of CAVE and HMD for visual stimulation in postural control research. AB - The control of balance of 20 healthy subjects under visual stimulation was quantitavely evaluated using a force platform. Three different visual stimuli were administered using a head-mounted display and CAVE. CAVE was significantly more effective in perturbing balance in one stimulus and HMD was significantly more effective in another. PMID- 15544311 TI - Virtual vision loss simulator. AB - Vision loss is common among people over the age of 65 and the condition is irreversible in most cases. Our simulator is a means to effectively and accurately inform people about the nature of common vision problems, initially limited to medically accurate simulations of cataract and macular degeneration. Using an eye tracker that continuously determines the subject's fixation point, a systematically degraded digital image displayed on an HMD can be a powerful educational tool. This article presents our Virtual Vision Loss (VVL) simulator and the developed simulation techniques based on calibration from 27 subjects. PMID- 15544312 TI - Reaction-time measurement and real-time data acquisition for neuroscientific experiments in virtual environments. AB - This paper describes an approach to neuroscientific reaction-time experiments and resulting methods for data processing under real-time constraints in virtual environments (VE). Immersive VEs produce huge amounts of data, which has to be processed in real-time to meet the interactivity requirement of VE. Furthermore, data is needed often with a timing resolution and even more crucial with an accuracy in the range of milliseconds. Unfortunately recent operating systems do not in general guarantee timing precision in this range. For these purposes we have researched possibilities for reaction-time measurement and real-time data acquisition. Our system enables neuroscientists to perform reaction-time experiments in platform-independent virtual environments. PMID- 15544313 TI - A preliminary study of presence in virtual reality training simulation for medical emergencies. AB - In this paper, a preliminary study of presence in a training simulation for medical emergency based on virtual reality is presented. We explore the influence of interaction mechanisms, as well as the complexity of behaviours in the subjective sense of presence. As expected, it has been found that as the type of interaction is more natural, and the patient behaviour modelling is more complex, the achieved sense of presence is greater. However our results also show that the degree of presence depends more upon the complexity of patient modelling than on how natural the interaction is. Hence, we postulate that a proper patient modelling could elicit a high degree of presence, even with traditional interaction mechanisms. PMID- 15544314 TI - An AR system with intuitive user interface for manipulation and visualization of 3D medical data. AB - We report on a stereoscopic video-see-through augmented reality system which we developed for medical applications. Our system allows interactive in-situ visualization of 3D medical imaging data. For high-quality rendering of the augmented scene we utilize the capabilities of the latest graphics card generations. Fast high-precision MPR generation ("multiplanar reconstruction") and volume rendering is realized with OpenGL 3D textures. We provide a tracked hand-held tool to interact with the medical imaging data in its actual location. This tool is represented as a virtual tool in the space of the medical data. The user can assign different functionality to it: select arbitrary MPR cross sections, guide a local volume rendered cube through the medical data, change the transfer function, etc. Tracking works in conjunction with retroreflective markers, which frame the workspace for head tracking respectively are attached to instruments for tool tracking. We use a single head-mounted tracking camera, which is rigidly fixed to the stereo pair of cameras that provide the live video view of the real scene. The user's spatial perception is based on stereo depth cues as well as on the kinetic depth cues that he receives with the viewpoint variations and the interactive data visualization. The AR system has a compelling real-time performance with 30 stereo-frames/second and exhibits no time lag between the video images and the augmenting graphics. Thus, the physician can interactively explore the medical imaging information in-situ. PMID- 15544315 TI - A haptic surgical simulator for the continuous curvilinear capsulorhexis procedure during cataract surgery. AB - This paper describes a technique for simulating the capsulorhexis procedure during cataract surgery. The continuous curvilinear capsulorhexis technique can be a difficult procedure for beginning ophthalmology surgeons. In the initial phase of tearing the tissue, the tear vector is tangential to the circumference of the tear circle. However, without the proper re-grasping of the flap of torn tissue close to the tear point, the tear vector angle quickly runs downhill possibly causing severe damage to the tissue. Novice surgeons tend to try to complete the capsulorhexis without the time consuming re-grasping of the tissue flap. Other factors such as anterior bowing of the lens diaphragm, patient age, and shallow anterior chambers add to the problematic nature of the procedure. Our capsulorhexis simulator models these various tear problems and anomalies to provide a training environment without the dangers of using live patients. PMID- 15544316 TI - Haptic rendering of tissue cutting with scissors. AB - Current surgical simulations that involve cutting with scissors lack display of forces "between the fingers" of the operator. The objective of this work is to develop a fast, realistic haptic rendering technique for scissor cutting that can be easily integrated into soft tissue models commonly used in surgical simulators. A virtual environment was created for combined graphic and haptic display of cutting. Two 2-dimensional deformable mass-spring-damper models were developed: The first is a large mesh that is used to calculate translational forces. The second is a smaller, one-element-thick mesh that is used to calculate cutting (angular) forces. This technique was implemented on the haptic scissors, a haptic interface that allows motion and force feedback in translation and cutting. PMID- 15544317 TI - Increasing face validity of a vascular interventional training system. AB - BACKGROUND: Many aspects of medical training take place on real patients in a live environment thus incurring risk. Apart from the obvious risks to patients there is the issue of X-ray exposure to both staff and trainees. Image quality used during interventional procedures is low to ensure minimum X-ray radiation dose. A virtual interventional system may be used to simulate the interventional cardiology training environment therefore reducing overall risk. The purpose of this project was to determine the appropriate image quality settings on a virtual training system to accurately simulate interventional cardiology and increase the face validity of the overall system. METHODS: Real image data was obtained from patients undergoing diagnostic interventional cardiology examinations in the Royal Victoria Hospital, Belfast. The images were obtained during catheter placement and contrast enhancement around the region of the heart. The diagnostic views were left anterior oblique, right anterior oblique, left lateral, caudal and the spider view. These five views were simulated on a virtual interventional training system (VIST, Mentice Medical Simulation AB, Sweden) by adjusting x-ray tube and table position at 20 different image quality settings. Signal to noise ratio and image contrast were measured for each of these views using soft tissue and vertebra. The results from the real image data and the VIST image data were compared. Calibration curves were compiled to determine the appropriate VIST image quality setting for each procedure. RESULTS: Average SNR ranged from 2.94 to 74.0 and IC ranged from 0.009 to 0.61 on the real image data. This indicates the very wide range of image quality encountered in interventional cardiology. Calibration curves for each view with the corresponding range of real patient image quality were produced. These calibration curves enabled the appropriate VIST image quality setting to be determined and therefore simulate the real examination image quality as closely as possible. CONCLUSIONS: The VIST system has been calibrated in terms of image quality for interventional cardiology examinations. This has increased the face validity of the system. PMID- 15544318 TI - An endoscopic sinus surgery training system for assessment of surgical skill. AB - The apprenticeship of procedures in endoscopic sinus surgery has several limitations, including potential risk for the patients and lack of feedback to the trainees. In this paper, we present a new surgical training system that combines a head dummy, force and position sensors: this system can be used to assess surgical skills and provide visual feedback to the trainees. PMID- 15544319 TI - Acquiring laparoscopic manipulative skills: a virtual tissue dissection training module. AB - Virtual environments for training manipulative skills in laparoscopic surgery are now well established as research areas. Tissue dissection however has not yet been adequately addressed. We have developed a virtual training module in which the task is to dissect a 3D tissue model using a simulated L-hook. Three metrics have been designed to assess performance. 1) Total deviation of actual cut path from the drawn path. 2) Total contact time between the instrument and tissue. 3) Contact discontinuity. The objective of the research was to design a basic tissue dissection module to train, in part, laparoscopic manipulative skills. PMID- 15544320 TI - Novel force resolver designs for a haptic surgery simulator. AB - This paper presents the results of an investigation into the design of low cost, four degree of freedom force/torque resolvers for use in compact haptic devices for surgical training. Two designs using piezo-resistive sensors are described and one using strain gauges. The advantages and disadvantages of each approach are stated and conclusions presented. PMID- 15544321 TI - Inverse thinking about double mutants of enzymes. AB - The quantitative effect of a second damaging mutation on a mutated enzyme may be additive, partially additive, synergistic, antagonistic, or absent, in the double mutant. Each of these five possible types of interactions has its own mechanistic explanation [Mildvan, A. S., Weber, D. J., and Kuliopulos, A. (1992) Arch. Biochem. Biophys. 294, 327-340]. Additive effects indicate independent functioning of the two residues in the process being studied, such as catalysis (k(cat)) or substrate binding (K(S)). Departures from additivity reflect interaction of the two residues. Thus, partial additivity indicates cooperativity, synergy indicates anticooperativity, and antagonism indicates opposing structural effects of the two mutations. No additional effects represent limiting cases of either partial additivity or antagonism. A significant conceptual simplification is achieved by applying inverse thinking, namely, by using the parameters of the double mutant rather than those of the wild-type enzyme as the reference point. To explain partially additive effects on k(cat), inverse thinking starts with the k(cat) of the double mutant. Restoring only one residue increases k(cat) by the factor A. Restoring only the other residue increases k(cat) by the factor B. Restoring both residues is shown to increase k(cat) by a factor greater than A x B, with the excess directly measuring the cooperativity. Similarly, inverse thinking provides simpler and more intuitive explanations of synergistic and antagonistic effects, as illustrated by specific examples. PMID- 15544322 TI - Computational prediction of structure, substrate binding mode, mechanism, and rate for a malaria protease with a novel type of active site. AB - The histo-aspartic protease (HAP) from the malaria parasite P. falciparum is one of several new promising targets for drug intervention. The enzyme possesses a novel type of active site, but its 3D structure and mechanism of action are still unknown. Here we use a combination of homology modeling, automated docking searches, and molecular dynamics/reaction free energy profile simulations to predict the enzyme structure, conformation of bound substrate, catalytic mechanism, and rate of the peptide cleavage reaction. We find that the computational tools are sufficiently reliable both for identifying substrate binding modes and for distinguishing between different possible reaction mechanisms. It is found that the favored pathway only involves direct participation by the catalytic aspartate, with the neighboring histidine providing critical stabilization (by a factor of approximately 10000) along the reaction. The calculated catalytic rate constant of about 0.1 s(-1) for a hexapeptide substrate derived from the alpha chain of human hemoglobin is in excellent agreement with experimental kinetic data for a similar peptide fragment. PMID- 15544324 TI - Engineering subtilisin into a fluoride-triggered processing protease useful for one-step protein purification. AB - Subtilisin was engineered into a highly specific, processing protease, and the subtilisin prodomain was coengineered into an optimized recognition sequence. This involved five steps. First, a robust subtilisin mutant was created, which could tolerate the subsequent mutations needed for high specificity. Second, the substrate binding pocket was mutated to increase its sequence selectivity. Third, the subtilisin prodomain was engineered to direct cleavage to the junction of any protein fused to it. Fourth, the active site of subtilisin was engineered to kinetically isolate binding and cleavage reactions. Finally, specific anions were identified to trigger the processing reaction, with fluoride ions being particularly useful. The ability to isolate the binding and processing steps with a triggering mechanism created a protease with a virtual on-off switch. This allowed column-immobilized processing subtilisin to be used as both the affinity ligand and processing protease for one-step purification of proteins. Fusion proteins tagged with the engineered prodomain can be bound to the column and washed free of contaminants. Cleavage can be triggered by the addition of fluoride to release the pure target protein. The column is then regenerated by stripping off the tightly bound prodomain at pH 2.1. Ten proteins have been purified to date by this method. PMID- 15544323 TI - Structural analysis of actinorhodin polyketide ketoreductase: cofactor binding and substrate specificity. AB - Aromatic polyketides are a class of natural products that include many pharmaceutically important aromatic compounds. Understanding the structure and function of PKS will provide clues to the molecular basis of polyketide biosynthesis specificity. Polyketide chain reduction by ketoreductase (KR) provides regio- and stereochemical diversity. Two cocrystal structures of actinorhodin polyketide ketoreductase (act KR) were solved to 2.3 A with either the cofactor NADP(+) or NADPH bound. The monomer fold is a highly conserved Rossmann fold. Subtle differences between structures of act KR and fatty acid KRs fine-tune the tetramer interface and substrate binding pocket. Comparisons of the NADP(+)- and NADPH-bound structures indicate that the alpha6-alpha7 loop region is highly flexible. The intricate proton-relay network in the active site leads to the proposed catalytic mechanism involving four waters, NADPH, and the active site tetrad Asn114-Ser144-Tyr157-Lys161. Acyl carrier protein and substrate docking models shed light on the molecular basis of KR regio- and stereoselectivity, as well as the differences between aromatic polyketide and fatty acid biosyntheses. Sequence comparison indicates that the above features are highly conserved among aromatic polyketide KRs. The structures of act KR provide an important step toward understanding aromatic PKS and will enhance our ability to design novel aromatic polyketide natural products with different reduction patterns. PMID- 15544325 TI - The role of Ca2+-coordinating residues of herring antifreeze protein in antifreeze activity. AB - The type II antifreeze protein of Atlantic herring (Clupea harengus harengus) requires Ca(2+) as a cofactor to inhibit the growth of ice crystals. On the basis of homology modeling with Ca(2+)-dependent lectin domains, five residues of herring antifreeze protein (hAFP) are predicted to be involved in Ca(2+) binding: Q92, D94, E99, N113, and D114. The role of E99, however, is less certain. A previous study on a double mutant EPN of hAFP suggested that the Ca(2+)-binding site of hAFP was the ice-binding site. However, it is possible that Ca(2+) might function distantly to affect ice binding. Site-directed mutagenesis was performed on the Ca(2+)-coordinating residues of hAFP in order to define the location of the ice-binding site and to explore the role of these residues in antifreeze activity. Properties of the mutants were investigated in terms of their structural integrity and antifreeze activity. Equilibrium dialysis analysis demonstrated that E99 is a Ca(2+)-coordinating residue. Moreover, proteolysis protection assay revealed that removal of Ca(2+) affected the conformation of the Ca(2+)-binding loop rather than the core structure of hAFP. This finding rules out the possibility that Ca(2+) might act at a distance via a conformational change to affect the function of hAFP. Substitutions at positions 99 and 114 resulted in severely reduced thermal hysteresis activity. These data indicate that the ice-binding site of hAFP is located at the Ca(2+)-binding site and the loop region defined by residues 99 and 114 is important for antifreeze activity. PMID- 15544326 TI - Epitope mapping of the phosphorylation motif of the HIV-1 protein Vpu bound to the selective monoclonal antibody using TRNOESY and STD NMR spectroscopy. AB - The conformational preferences of a 22-amino acid peptide (LIDRLIERAEDpSGNEpSEGEISA) that mimics the phosphorylated HIV-1-encoded virus protein U (Vpu) antigen have been investigated by NMR spectroscopy. Degradation of HIV receptor CD4 by the proteasome, mediated by the HIV-1 protein Vpu, is crucial for the release of fully infectious virions. Phosphorylation of Vpu at sites Ser52 and Ser56 on the DSGXXS motif is required for the interaction of Vpu with the ubiquitin ligase SCF(beta)(-TrCP) which triggers CD4 degradation by the proteasome. This motif is conserved in several signaling proteins known to be degraded by the proteasome. The interaction of the P-Vpu(41-62) peptide with its monoclonal antibody has been studied by transferred nuclear Overhauser effect NMR spectroscopy (TRNOESY) and saturation transfer difference NMR (STD NMR) spectroscopy. The peptide was found to adopt a bend conformation upon binding to the antibody; the peptide residues (Asp51-pSer56) forming this bend are recognized by the antibody as demonstrated by STD NMR experiments. The three dimensional structure of P-Vpu(41-62) in the bound conformation was determined by TRNOESY spectra; the peptide adopts a compact structure in the presence of mAb with formation of several bends around Leu45 and Ile46 and around Ile60 and Ser61, with a tight bend created by the DpS(52)GNEpS(56) motif. STD NMR studies provide evidence for the existence of a conformational epitope containing tandem repeats of phosphoserine motifs. The peptide's epitope is predominantly located in the large bend and in the N-terminal segment, implicating bidentale association. These findings are in excellent agreement with a recently published NMR structure required for the interaction of Vpu with the SCF(beta)(-TrCP) protein. PMID- 15544327 TI - A copper protein and a cytochrome bind at the same site on bacterial cytochrome c peroxidase. AB - Pseudoazurin binds at a single site on cytochrome c peroxidase from Paracoccus pantotrophus with a K(d) of 16.4 microM at 25 degrees C, pH 6.0, in an endothermic reaction that is driven by a large entropy change. Sedimentation velocity experiments confirmed the presence of a single site, although results at higher pseudoazurin concentrations are complicated by the dimerization of the protein. Microcalorimetry, ultracentrifugation, and (1)H NMR spectroscopy studies in which cytochrome c550, pseudoazurin, and cytochrome c peroxidase were all present could be modeled using a competitive binding algorithm. Molecular docking simulation of the binding of pseudoazurin to the peroxidase in combination with the chemical shift perturbation pattern for pseudoazurin in the presence of the peroxidase revealed a group of solutions that were situated close to the electron transferring heme with Cu-Fe distances of about 14 A. This is consistent with the results of (1)H NMR spectroscopy, which showed that pseudoazurin binds closely enough to the electron-transferring heme of the peroxidase to perturb its set of heme methyl resonances. We conclude that cytochrome c550 and pseudoazurin bind at the same site on the cytochrome c peroxidase and that the pair of electrons required to restore the enzyme to its active state after turnover are delivered one-by-one to the electron-transferring heme. PMID- 15544328 TI - Phospholipase A2 as a target protein for nonsteroidal anti-inflammatory drugs (NSAIDS): crystal structure of the complex formed between phospholipase A2 and oxyphenbutazone at 1.6 A resolution. AB - Phospholipase A(2) (PLA(2); EC 3.1.1.4) is a key enzyme involved in the production of proinflammatory mediators known as eicosanoids. The binding of the substrate to PLA(2) occurs through a well-formed hydrophobic channel. To determine the viability of PLA(2) as a target molecule for the structure-based drug design against inflammation, arthritis, and rheumatism, the crystal structure of the complex of PLA(2) with a known anti-inflammatory compound oxyphenbutazone (OPB), which has been determined at 1.6 A resolution. The structure has been refined to an R factor of 0.209. The structure contains 1 molecule each of PLA(2) and OPB with 2 sulfate ions and 111 water molecules. The binding studies using surface plasmon resonance show that OPB binds to PLA(2) with a dissociation constant of 6.4 x 10(-8) M. The structure determination has revealed the presence of an OPB molecule at the binding site of PLA(2). It fits well in the binding region, thus displaying a high level of complementarity. The structure also indicates that OPB works as a competitive inhibitor. A large number of hydrophobic interactions between the enzyme and the OPB molecule have been observed. The hydrophobic interactions involving residues Tyr(52) and Lys(69) with OPB are particularly noteworthy. Other residues of the hydrophobic channel such as Leu(3), Phe(5), Met(8), Ile(9), and Ala(18) are also interacting extensively with the inhibitor. The crystal structure clearly reveals that the binding of OPB to PLA(2) is specific in nature and possibly suggests that the basis of its anti-inflammatory effects may be due to its binding to PLA(2) as well. PMID- 15544329 TI - Oncogenic Dbl, Cdc42, and p21-activated kinase form a ternary signaling intermediate through the minimum interactive domains. AB - Activation of many Rho family GTPase pathways involves the signaling module consisting of the Dbl-like guanine nucleotide exchange factors (GEFs), the Rho GTPases, and the Rho GTPase specific effectors. The current biochemical model postulates that the GEF-stimulated GDP/GTP exchange of Rho GTPases leads to the active Rho-GTP species, and subsequently the active Rho GTPases interact with and activate the effectors. Here we report an unexpected finding that the Dbl oncoprotein, Cdc42 GTPase, and PAK1 can form a complex through their minimum functional motifs, i.e., the Dbl-homolgy (DH) and Pleckstrin-homology domains of Dbl, Cdc42, and the PBD domain of PAK1. The Dbl-Cdc42-PAK1 complex is sensitive to the nucleotide-binding state of Cdc42 since either dominant negative or constitutively active Cdc42 readily disrupts the ternary binding interaction. The complex formation depends on the interactions between the DH domain of Dbl and Cdc42 and between Cdc42 and the PBD domain of PAK1 and can be reconstituted in vitro by using the purified components. Furthermore, the Dbl-Cdc42-PAK1 ternary complex is active in generating signaling output through the activated PAK1 kinase in the complex. The GEF-Rho-effector ternary intermediate is also found in other Dbl-like GEF, Rho GTPase, and effector interactions. Finally, PAK1, through the PDB domain, is able to accelerate the GEF-induced GTP loading onto Cdc42. These results suggest that signal transduction through Cdc42 and possibly other Rho family GTPases could involve tightly coupled guanine nucleotide exchange and effector activation mechanisms and that Rho GTPase effector may have a feedback regulatory role in the Rho GTPase activation. PMID- 15544330 TI - Molecular functions of conserved aspects of the GHMP kinase family. AB - The sequences and three-dimensional structures of the galactokinase, homoserine kinase, mevalonate kinase, and phosphomevalonate kinase (GHMP) family were compared to identify highly conserved surface residues. The functions of these solvent-accessible residues were assessed by determining the effects of their substitution, via mutagenesis, on the initial-rate parameters of a representative member of the GHMP kinase family, phosphomevalonate kinase from Streptococcus pneumoniae. What emerges from this study is a profile of the conserved surface linked functions of the family. Certain substitutions produce highly selective effects on the steady-state affinity of a particular substrate, while one residue, Asp150, appears to be a pure k(cat) effector. Substitutions elsewhere affect multiple initial-rate parameters with varying, and sometimes compensatory, patterns. An alpha-helix that repositions during catalysis was substituted along its length to assess how its different segments contribute to catalysis-the substrate-proximal edge of the helix affects ATP recognition and k(cat), while the distal edge affects recognition of both substrates without affecting turnover. GHMP kinase mutations at the conserved surface residues corresponding to Ser291 and Ala293 in phosphomevalonate kinase are linked to mevalonic acid deficiency, which can lead to early fatality, and galactokinase deficiency, which causes cataracts. Our results suggest that the molecular basis for this particular galactokinase deficiency is an increase in the K(m) for galactose. PMID- 15544331 TI - Chemokine-binding specificity of soluble chemokine-receptor analogues: identification of interacting elements by chimera complementation. AB - The specificity of chemokine-receptor interactions plays a central role in the regulation of leukocyte migration in inflammatory responses. Herein, we describe a soluble mimic of CC chemokine receptor 2 (CCR2), dubbed CROSS-N(2)E3(2), which incorporates the N-terminal region (N) and third extracellular loop (E3) elements of CCR2 displayed on the surface of a soluble protein scaffold. CROSS-N(2)E3(2) binds to the CCR2 ligand monocyte chemoattractant protein-1 (MCP-1) with a dissociation equilibrium constant of 1.1 +/- 0.1 microM but does not bind to the cognate chemokines of the receptor CCR3 (eotaxin-1, -2, and -3). Similarly, a soluble analogue of CCR3 (CROSS(5)-N(3)E3(3)) binds to eotaxin-1, -2, and -3 but not to MCP-1. Thus, these receptor analogues have the same specificity as the natural receptors. Using soluble proteins containing N and E3 elements from different receptors (CROSS-N(2)E3(3) and CROSS-N(3)E3(2)), we demonstrate that both receptor elements are required for optimal binding to the cognate chemokines. In addition, we report the binding affinities of all four CROSS proteins to a panel of two wild-type and six chimeric chemokines. These complementation studies indicate the regions of the chemokines that interact with each element of the receptors, allowing us to deduce the orientations of the receptor extracellular elements relative to the bound chemokines. PMID- 15544332 TI - Role of base stacking and sequence context in the inhibition of yeast DNA polymerase eta by pyrene nucleotide. AB - The Y family DNA polymerase yeast pol eta inserts pyrene deoxyribose monophosphate (dPMP) in preference to A opposite an abasic site, the 3'-T of a thymine dimer, and a normal T with almost equal efficiency. In contrast, pol A family polymerases such as Klenow fragment and T7 DNA polymerase only insert dPMP efficiently opposite an abasic site and the 3'-T of a thymine dimer but not opposite undamaged DNA. Pyrene nucleotide is also an efficient chain-terminating inhibitor of DNA synthesis by pol eta but not by Klenow fragment or T7 DNA polymerase. To better understand the origin of the efficiency and sequence specificity of dPMP insertion by pol eta, the kinetics of dPMP insertion opposite various templates have been determined. In one sequence context, the efficiency of dPMP insertion increases 4.6-fold opposite G < A << T < C, suggesting that the templating nucleotide modulates dPMP insertion efficiency by having to destack prior to dPTP binding. The efficiency of insertion of dPMP opposite T in the same sequence context increases 7-fold for primers terminating in G < A < C < T and is similar to that observed for nontemplated blunt-end extension, suggesting that stacking interactions between the pyrene and the primer terminus are also important. On heterogeneous templates, the average selectivity for dPMP insertion relative to the complementary dNMP decreases in the order of dAMP > dGMP > dTMP > dCMP, from a high of 5.8 when dAMP is to be inserted following a T to a low of 0.5 when dCMP is to be inserted following a C. The relative preference for dPMP insertion at a given site can be largely explained by the energetic cost of destacking the templating base and stacking of pyrene nucleotide relative to that of stacking and base pairing the complementary nucleotide. Thus, pyrene nucleotide represents a novel class of nucleotide-based chain-terminating DNA synthesis inhibitors whose base portion consists of a hydrophobic, non-hydrogen bonding, base-pair mimic. PMID- 15544333 TI - Dynamics inherent in helix 27 from Escherichia coli 16S ribosomal RNA. AB - The original interpretation of a series of genetic studies suggested that the highly conserved Escherichia coli 16S ribosomal RNA helix 27 (H27) adopts two alternative secondary structure motifs, the 885 and 888 conformations, during each cycle of amino acid incorporation. Recent crystallographic and genetic evidence has called this hypothesis into question. To ask whether a slippery sequence such as that of H27 may harbor inherent conformational dynamics, we have designed a series of model RNAs based on E. coli H27 for in vitro physicochemical studies. One-dimensional (1)H NMR spectroscopy demonstrates that both the 885 and 888 conformations are occupied to approximately the same extent (f(888) = 0.427 +/- 0.04) in the native H27 sequence at low pH (6.4) and low ionic strength (50 mM NaCl). UV irradiation assays conducted under conditions analogous to those used for assays of ribosomal function (pH 7.5 and 20 mM MgCl(2)) suggest that nucleotides 892 and 905, which are too far apart in the known 885 crystal structures, can approach each other closely enough to form an efficient cross link. The use of a fluorescence resonance energy transfer (FRET)-labeled RNA together with a partially complementary DNA oligonucleotide that induces a shift to the 888 conformation shows that H27 interchanges between the 885 and 888 conformations on the millisecond time scale, with an equilibrium constant of 0.33 +/-0.12. FRET assays also show that tetracycline interferes with the induced shift to the 888 conformation, a finding that is consistent with crystallographic localization of tetracycline bound to the 885 conformation of H27 in the 30S ribosomal subunit. Taken together, our data demonstrate the innate tendency of an isolated H27 to exist in a dynamic equilibrium between the 885 and 888 conformations. This begs the question of how these inherent structural dynamics are suppressed within the context of the ribosome. PMID- 15544334 TI - Denaturant-induced unfolding of the acetyl-esterase from Escherichia coli. AB - The stability of acetyl-esterase, Aes, from Escherichia coli against the denaturing action of urea and guanidine hydrochloride, GuHCl, has been investigated by means of circular dichroism and fluorescence measurements. The urea-induced unfolding curves show a single inflection point at 6.2 M urea, whereas the GuHCl-induced curves show two inflection points at 1.4 and 3.1 M GuHCl. The unfolding process is reversible with both urea and GuHCl. These results, together with similar experimental data on the mutant form V20D-Aes, suggest the presence of two domains in the Aes structure, which unfold more or less independently depending on the denaturant used. This is also supported by a 3D model obtained by homology modeling using the structure of brefeldine as a template. The effect of NaCl on the urea-induced unfolding curves of the enzyme has also been investigated. PMID- 15544335 TI - Changes in the functional and structural properties of the Mn cluster induced by replacing the side group of the C-terminus of the D1 protein of photosystem II. AB - A free alpha-COO(-) in the C-terminal alanine-344 (Ala344) in the D1 protein of photosystem II is thought to be responsible for ligating the Mn cluster. The effects of the side group of the C-terminus of the D1 protein on the functional and structural properties of the oxygen-evolving complex (OEC) were comprehensively studied by replacing Ala344 with glycine (Gly), valine (Val), aspartate (Asp), or asparagine (Asn). All the mutants grew photoautotrophically under low-light conditions with lower O(2) evolution activity depending on the mutants when compared with the activity of the control wild type. The Gly-, Asp-, and Asn-substituted mutants did not grow under high-light conditions, while the Val-substituted mutant grew even under the high-light conditions. S(2)-state thermoluminescence bands appeared at slightly elevated temperatures when compared with those of the wild type in the Asp- and Gly-substituted mutants, but at almost normal temperatures in the Val- and Asn-substituted mutants. The oxygen evolving core particles isolated from the mutants showed little change in protein composition. The Gly-, Asp-, and Asn-substituted core particles exhibited low temperature electron spin resonance (ESR) spectra with reduced S(2) multiline and enhanced g = 4.1 ESR signals, while the Val-substituted particles showed a spectrum similar to that of the control particles. Mid-frequency Fourier transform infrared difference spectra showed distinctive changes in several bands arising from the putative carboxylate ligands for the Mn cluster in all substituted particles, but the bands for the putative C-terminal alpha carboxylate did not seem to change in the substituted spectra. The changes induced by the Asp and Asn substitution resembled each other except for the amide I region, and showed some similarity to those induced by the Gly substitution in the symmetric carboxylate stretching region. The results were interpreted to mean that similar types of changes of the carboxylate ligands are induced by these substitutions. The band from a putative histidine ligand for the Mn cluster was similarly affected in the Gly-, Asp-, and Asn-substituted spectra, but not in the Val-substituted spectrum. Notably, marked changes in the amide I, amide II, and carboxylate bands were observed in the Val-substituted spectrum, which was different from the Gly-, Asp-, and Asn-substituted spectra. The results indicated that the structural perturbations induced by the Val substitution include large changes of the protein backbone and are considerably different from those induced by the other substitutions. Possible amino acid ligands participating in the changes deduced by Ala344 replacement in the D1 C-terminal and the effects of the changes of the side group on these ligands were considered on the basis of the available X-ray model of the OEC. PMID- 15544336 TI - Modulation of human 5-lipoxygenase activity by membrane lipids. AB - Mammalian 5-lipoxygenase (5-LO) catalyzes the conversion of arachidonic acid (AA) to leukotrienes, potent inflammatory mediators. 5-LO is activated by a Ca(2+) mediated translocation to membranes, and demonstrates the characteristic features of interfacially activated enzymes, yet the mechanism of membrane binding of 5-LO is not well understood. In an attempt to understand the mechanism of lipid mediated activation of 5-LO, we have studied the effects of a large set of lipids on human recombinant 5-LO activity, as well as mutual structural effects of 5-LO and membranes. In the presence of 0.35 mM phosphatidylcholine (PC) and 0.2 mM Ca(2+), there was substrate inhibition at >100 microM AA. Data analysis at low AA concentrations yielded the following: K(m) approximately 103 microM and k(cat) approximately 56 s(-1). 5-LO activity was supported by PC more than by any other lipid tested except for a cationic lipid, which was more stimulatory than PC. Binding of 5-LO to zwitterionic and acidic membranes was relatively weak; the extent of binding increased 4-8 times in the presence of Ca(2+), whereas binding to cationic membranes was stronger and essentially Ca(2+)-independent. Polarized attenuated total reflection infrared experiments implied that 5-LO binds to membranes at a defined orientation with the symmetry axis of the putative N terminal beta-barrel tilted approximately 45 degrees from the membrane normal. Furthermore, membrane binding of 5-LO resulted in dehydration of the membrane surface and was paralleled with stabilization of the structures of both 5-LO and the membrane. Our results provide insight into the understanding of the effects of membrane surface properties on 5-LO-membrane interactions and the interfacial activation of 5-LO. PMID- 15544337 TI - Thermodynamics of aminoglycoside binding to aminoglycoside-3'-phosphotransferase IIIa studied by isothermal titration calorimetry. AB - The aminoglycoside-3'-phosphotransferase IIIa [APH(3')-IIIa] phosphorylates aminoglycoside antibiotics and renders them ineffective against bacteria. APH(3') IIIa is the most promiscuous aminoglycoside phosphotransferase enzyme, and it modifies more than 10 different aminoglycoside antibiotics. A wealth of information exists about the enzyme; however, thermodynamic properties of enzyme aminoglycoside complexes are still not known. This study describes the determination of the thermodynamic parameters of the binary enzyme-aminoglycoside and the ternary enzyme-metal-ATP-aminoglycoside complexes of structurally related aminoglycosides using isothermal titration calorimetry. Formation of the binary enzyme-aminoglycoside complexes is enthalpically driven and exhibits a strongly disfavored entropic contribution. Formation of the ternary enzyme-metal-ATP aminoglycoside complexes yields much smaller negative DeltaH values and more favorable entropic contributions. The presence of metal-ATP generally increases the affinity of aminoglycosides to the enzyme. This is consistent with the kinetic mechanism of the enzyme in which ordered binding of substrates occurs. However, the observed DeltaH values neither correlate with kinetic parameters k(cat), K(m), and k(cat)/K(m) nor correlate with the molecular size of the substrates. Comparison of the thermodynamic properties of the complexes formed by structurally similar aminoglycosides indicated that the 2'- and the 6'-amino groups of the substrates are involved in binding to the enzyme. Thermodynamic properties of the complexes formed by aminoglycosides differing only at the 3' hydroxyl group suggested that the absence of this group does not alter the thermodynamic parameters of the ternary APH(3')-IIIa-metal-ATP-aminoglycoside complex. Our results also indicate that protonation of ligand and protein ionizable groups is coupled to the complex formation between aminoglycosides and APH(3')-IIIa. Comparison of DeltaH values for different aminoglycoside-enzyme complexes indicates that enzyme and substrates undergo significant conformational changes in complex formation. PMID- 15544338 TI - Evidence for increased local flexibility in psychrophilic alcohol dehydrogenase relative to its thermophilic homologue. AB - The psychrophilic alcohol dehydrogenase (psADH) cloned from Antarctic Moraxella sp. TAE123 exhibits distinctive catalytic parameters in relation to the homologous thermophilic alcohol dehydrogenase (htADH) from Bacillus stearothermophilus LLD-R. Amide hydrogen-deuterium (H/D) exchange studies using Fourier-transformed infrared (FTIR) spectroscopy and mass spectrometry (MS) were conducted to investigate whether the differences are caused by variation in either global or regional protein flexibility. The FTIR H/D exchange study suggested that psADH does not share similar global flexibility with htADH at their physiologically relevant temperatures as has been predicted by the "corresponding state" hypothesis. However, the MS H/D exchange study revealed a more complicated picture concerning the flexibility of the two homologous enzymes. Analysis of the deuteration and exchange rates of protein-derived peptides suggested that only some functionally important regions in psADH that are involved in substrate and cofactor binding exhibit greater flexibility compared to htADH at low temperature (10 degrees C). These observations strongly suggest that variable conformational flexibility between the two protein forms is a local phenomenon, and that global H/D exchange measurement by FTIR can be misleading and should be used with discretion. These results are supportive of the idea that functionally important local flexibility can be uncoupled from global thermal stability. The structural factors underlying the differences in local protein flexibility and catalysis between htADH and psADH are discussed in conjunction with results from crystallographic and fluorescence spectroscopy studies. PMID- 15544339 TI - The redox behavior of the heme in cystathionine beta-synthase is sensitive to pH. AB - Human cystathionine beta-synthase (CBS) is a unique pyridoxal-5'-phosphate dependent enzyme in which heme is also present as a cofactor. Because the function of heme in this enzyme has yet to be elucidated, the study presented herein investigated possible relationships between the chemistry of the heme and the strong pH dependence of CBS activity. This study revealed, via study of a truncation variant, that the catalytic core of the enzyme governs the pH dependence of the activity. The heme moiety was found to play no discernible role in regulating CBS enzyme activity by sensing changes in pH, because the coordination sphere of the heme is not altered by changes in pH over a range of pH 6-9. Instead, pH was found to control the equilibrium amount of ferric and ferrous heme present after reaction of CBS with one-electron reducing agents. A variety of spectroscopic techniques, including resonance Raman, magnetic circular dichroism, and electron paramagnetic resonance, demonstrated that at pH 9 Fe(II) CBS is dominant while at pH 6 Fe(III) CBS is favored. At low pH, Fe(II) CBS forms transiently but reoxidizes by an apparent proton-gated electron-transfer mechanism. Regulation of CBS activity by the iron redox state has been proposed as the role of the heme moiety in this enzyme. Given that the redox behavior of the CBS heme appears to be controlled by pH, interplay of pH and oxidation state effects must occur if CBS activity is redox regulated. PMID- 15544340 TI - Sulfide dehydrogenase activity of the monomeric flavoprotein SoxF of Paracoccus pantotrophus. AB - Flavocytochrome c-sulfide dehydrogenases (FCSDs) are complexes of a flavoprotein with a c-type cytochrome performing hydrogen sulfide-dependent cytochrome c reduction in vitro. The amino acid sequence analysis revealed that the phylogenetic relationship of different flavoproteins reflected the relationship of sulfur-oxidizing bacteria. The flavoprotein SoxF of Paracoccus pantotrophus is 29-67% identical to the flavoprotein subunit of FCSD of phototrophic sulfur oxidizing bacteria. Purification of SoxF yielded a homogeneous emerald-green monomeric protein of 42 797 Da. SoxF catalyzed sulfide-dependent horse heart cytochrome c reduction at the optimum pH of 6.0 with a k(cat) of 3.9 s(-1), a K(m) of 2.3 microM for sulfide, and a K(m) of 116 microM for cytochrome c, as determined by nonlinear regression analysis. The yield of 1.9 mol of cytochrome c reduced per mole of sulfide suggests sulfur or polysulfide as the product. Sulfide dehydrogenase activity of SoxF was inhibited by sulfur (K(i) = 1.3 microM) and inactivated by sulfite. Cyanide (1 mM) inhibited SoxF activity at pH 6.0 by 25% and at pH 8.0 by 92%. Redox titrations in the infrared spectral range from 1800 to 1200 cm(-1) and in the visible spectral range from 400 to 700 nm both yielded a midpoint potential for SoxF of -555 +/- 10 mV versus Ag/AgCl at pH 7.5 and -440 +/- 20 mV versus Ag/AgCl at pH 6.0 (-232 mV versus SHE') and a transfer of 1.9 electrons. Electrochemically induced FTIR difference spectra of SoxF as compared to those of free flavin in solution suggested a strong cofactor interaction with the apoprotein. Furthermore, an activation/variation of SoxF during the redox cycles is observed. This is the first report of a monomeric flavoprotein with sulfide dehydrogenase activity. PMID- 15544341 TI - The folding state of the lumenal loop determines the thermal stability of light harvesting chlorophyll a/b protein. AB - The major light-harvesting protein of photosystem II (LHCIIb) is the most abundant chlorophyll-binding protein in the thylakoid membrane. It contains three membrane-spanning alpha helices; the first and third one closely interact with each other to form a super helix, and all three helices bind most of the pigment cofactors. The protein loop domains connecting the alpha helices also play an important role in stabilizing the LHCIIb structure. Single amino acid exchanges in either loop were found to be sufficient to significantly destabilize the complex assembled in vitro [Heinemann, B., and Paulsen, H. (1999) Biochemistry 38, 14088-14093. Mick, V., Eggert, K., Heinemann, B., Geister, S., and Paulsen, H (2004) Biochemistry 43, 5467-5473]. This work presents an analysis of such point mutations in the lumenal loop with regard to the extent and nature of their effect on LHCIIb stability to obtain detailed information on the contribution of this loop to stabilizing the complex. Most of the mutant proteins yielded pigment protein complexes if their reconstitution and/or isolation was performed under mild conditions; however, the yields were significantly different. Several mutations in the vicinity of W97 in the N-proximal section of the loop gave low reconstitution yields even under very mild conditions. This confirms our earlier notion that W97 may be of particular relevance in stabilizing LHCIIb. The same amino acid exchanges accelerated thermal complex dissociation in the absence of lithium dodecyl sulfate (LDS) and raised the accessibility of the lumenal loop to protease; both effects were well correlated with the reduction in reconstitution yields. We conclude that a detachment of the lumenal loop is a possible first step in the dissociation of LHCIIb. Dramatically reduced complex yields in the presence but not in the absence of LDS were observed for some but not all mutants, particularly those near the C-proximal end of the loop. We conclude that complex stabilities in the absence and in the presence of LDS do not correlate and most likely are determined by different structural characteristics, at least in LHCIIb but maybe also in other membrane proteins. PMID- 15544342 TI - EpoK, a cytochrome P450 involved in biosynthesis of the anticancer agents epothilones A and B. Substrate-mediated rescue of a P450 enzyme. AB - The epothilones are a new class of highly promising anticancer agents with a mode of action akin to that of paclitaxel but with distinct advantages over that drug. The principal natural compounds are epothilones A and B, which have an epoxide in the macrocyclic lactone ring, and C and D, which have a double bond instead of the epoxide group. The epoxidation of epothilones C and D to A and B, respectively, is mediated by EpoK, a cytochrome P450 enzyme encoded in the epothilone gene cluster. Here we report high-yield expression of EpoK, characterization of the protein, demonstration that the natural substrate can prevent-and even reverse-denaturation of the protein, identification of ligands and surrogate substrates, development of a high-throughput fluorescence activity assay based on the H(2)O(2)-dependent oxidation of 7-ethoxy-4 trifluoromethylcoumarin, and identification of effective inhibitors of the enzyme. These results will facilitate improvements in the yields of epothilones C and D and the engineering of EpoK to prepare novel epothilone analogues. Furthermore, the finding that the denatured enzyme is rescued by the substrate offers a potential paradigm for control of the P450 catalytic function. PMID- 15544343 TI - Mechanism of formation of the complex between transferrin and bismuth, and interaction with transferrin receptor 1. AB - The kinetics and thermodynamics of Bi(III) exchange between bismuth mononitrilotriacetate (BiL) and human serum transferrin as well as those of the interaction between bismuth-loaded transferrin and transferrin receptor 1 (TFR) were investigated at pH 7.4-8.9. Bismuth is rapidly exchanged between BiL and the C-site of human serum apotransferrin in interaction with bicarbonate to yield an intermediate complex with an effective equilibrium constant K(1) of 6 +/- 4, a direct second-order rate constant k(1) of (2.45 +/- 0.20) x 10(5) M(-1) s(-1), and a reverse second-order rate constant k(-1) of (1.5 +/- 0.5) x 10(6) M(-1) s( 1). The intermediate complex loses a single proton with a proton dissociation constant K(1a) of 2.4 +/- 1 nM to yield a first kinetic product. This product then undergoes a modification in its conformation followed by two proton losses with a first-order rate constant k(2) = 25 +/- 1.5 s(-1) to produce a second kinetic intermediate, which in turn undergoes a last modification in the conformation to yield the bismuth-saturated transferrin in its final state. This last process rate-controls Bi(III) uptake by the N-site of the protein and is independent of the experimental parameters with a constant reciprocal relaxation time tau(3)(-1) of (3 +/- 1) x 10(-2) s(-1). The mechanism of bismuth uptake differs from that of iron and probably does not involve the same transition in conformation from open to closed upon iron uptake. The interaction of bismuth loaded transferrin with TFR occurs in a single very fast kinetic step with a dissociation constant K(d) of 4 +/- 0.4 microM, a second-order rate constant k(d) of (2.2 +/- 1.5) x 10(8) M(-1) s(-1), and a first-order rate constant k(-d) of 900 +/- 400 s(-1). This mechanism is different from that observed with the ferric holotransferrin and implies that the interaction between TFR and bismuth-loaded transferrin probably takes place on the helical domain of the receptor which is specific for the C-site of transferrin and HFE. The relevance of bismuth incorporation by the transferrin receptor-mediated iron acquisition pathway is discussed. PMID- 15544344 TI - The exclusion of glycine betaine from anionic biopolymer surface: why glycine betaine is an effective osmoprotectant but also a compatible solute. AB - Paradoxically, glycine betaine (N,N,N-trimethyl glycine; GB) in vivo is both an effective osmoprotectant (efficient at increasing cytoplasmic osmolality and growth rate) and a compatible solute (without deleterious effects on biopolymer function, including stability and activity). For GB to be an effective osmoprotectant but not greatly affect biopolymer stability, we predict that it must interact very differently with folded protein surface than with that exposed in unfolding. To test this hypothesis, we quantify the preferential interaction of GB with the relatively uncharged surface exposed in unfolding the marginally stable lacI helix-turn-helix (HTH) DNA binding domain using circular dichroism and with the more highly charged surfaces of folded hen egg white lysozyme (HEWL) and bovine serum albumin (BSA) using all-gravimetric vapor pressure osmometry (VPO) and compare these results with results of VPO studies (Hong et al. (2004), Biochemistry, 43, 14744-14758) of the interaction of GB with polyanionic duplex DNA. For these four biopolymer surfaces, we observe that the extent of exclusion of GB per unit of biopolymer surface area increases strongly with increasing fraction of anionic oxygen (protein carboxylate or DNA phosphate) surface. In addition, GB is somewhat more excluded from the surface exposed in unfolding the lacI HTH and from the folded surface of HEWL than expected from their small fraction of anionic surface, consistent with moderate exclusion of GB from polar amide surface, as predicted by the osmophobic model of protein stability (Bolen and Baskakov (2001) J. Mol. Biol. 310, 955-963). Strong exclusion of GB from anionic surface explains how it can be both an effective osmoprotectant and a compatible solute; analysis of this exclusion yields a lower bound on the hydration of anionic protein carboxylate surface of two layers of water (>or=0.22 H(2)O A(-)(2)). PMID- 15544345 TI - Preferential interactions of glycine betaine and of urea with DNA: implications for DNA hydration and for effects of these solutes on DNA stability. AB - Interactions of the solutes glycine betaine (GB) and urea with mononucleosomal calf thymus DNA in aqueous salt solutions are characterized by vapor pressure osmometry (VPO). Analysis of osmolality as a function of solute and DNA concentration yields the effect of the solute on the chemical potential, mu(2), of the DNA. Although both GB and urea generally are nucleic acid denaturants and therefore must interact favorably with the nucleic acid surface exposed upon melting, VPO demonstrates that neither interacts favorably with duplex DNA. Addition of GB greatly increases mu(2) of DNA, indicating that the average local concentration of GB in the vicinity of the double helix is much less than its bulk concentration. By contrast, addition of urea has almost no effect on mu(2) of duplex DNA, indicating that the average local concentration of urea in the vicinity of duplex DNA is almost the same as in bulk solution. Qualitatively, we conclude that the nonuniform distribution of GB occurs primarily because duplex DNA and GB prefer to interact with water rather than with each other. Comparison with thermodynamic data for the interaction of GB with various protein surfaces (Felitsky et al., Biochemistry, 43, 14732-14743) shows that GB is excluded primarily from anionic DNA surface and that the hydration of anionic DNA phosphate oxygen surface (>or approximately 17 H(2)O per nucleotide or >or approximately 0.22 H(2)O A(-)(2)) involves at least two layers of water. From analysis of literature data for effects of urea and of GB on DNA melting, we propose that urea is an effective nonspecific nucleic acid denaturant because of its favorable interactions with the polar amide-like surface of G, C, and especially T or U bases exposed in denaturation, whereas GB is a specific GC denaturant because of its favorable interaction with G and/or C surface in the single-stranded state. PMID- 15544346 TI - Elongation factor Ts from the Antarctic eubacterium Pseudoalteromonas haloplanktis TAC 125: biochemical characterization and cloning of the encoding gene. AB - The elongation factor Ts was isolated from the psychrophilic Antarctic eubacterium Pseudoalteromonas haloplanktis TAC 125 strain (PhEF-Ts), and its functional properties were studied. At 0 degrees C PhEF-Ts enhanced the [(3)H]GDP/GDP exchange rate on the preformed PhEF-Tu.[(3)H]GDP complex by 2 orders of magnitude even at very low Tu:Ts ratio, by lowering the energy of activation of the exchange reaction. PhEF-Ts is a monomeric protein, and in solution it forms a stable dimeric complex with PhEF-Tu. The PhEF-Ts encoding gene was cloned and sequenced. Its structural organization was similar to that of Escherichia coli because it showed at its 5' end the gene encoding the ribosomal protein S2. The translated amino acid sequence had a calculated molecular weight of 30762, and showed a high sequence identity with E. coli (68%) and Thermus thermophilus (44%) EF-Ts. The PhEF-Ts primary structure contains well-preserved almost all the amino acid residues interacting at the interfaces of the E. coli EF-Ts.EF-Tu complex. Finally, the high concentration of PhEF-Ts in this psychrophilic eubacterium might represent an adaptive tool to ensure an efficient nucleotide exchange even at low temperature. PMID- 15544347 TI - Lipid modulation of the activity of diacylglycerol kinase alpha- and zeta isoforms: activation by phosphatidylethanolamine and cholesterol. AB - Diacylglycerol kinase (DGK) isoforms alpha and zeta were extracted from transfected cells that overexpressed these enzymes. We determined the lipid dependence of the binding of these isoforms to liposomes. The modulation by lipid of the rate of phosphorylation of diacylglycerol by these enzymes was also measured. Incorporation of phosphatidylethanolamine into the liposomes resulted in an increased partitioning of both isoforms of DGK to the membrane as well as an increased catalytic rate. We demonstrate that the increased catalytic rate is a consequence of both increased portioning of the enzyme to the membrane and increased catalytic activity of the membrane-bound form. DGKalpha, a calcium dependent isoform, can be activated in a calcium-independent fashion in the presence of phosphatidylethanolamine. Similar effects are observed with cholesterol. In contrast, sphingomyelin inhibits the activity of both isoforms of DGK. Our results demonstrate that the translocation to membranes and activity of DGKalpha and DGKzeta are modulated by the composition and properties of the membrane. The enzymes are activated by the presence of lipids that promote the formation of inverted phases. However, the promotion of negative curvature is not the sole factor contributing to the lipid effects on enzyme binding and activity. A truncated form of DGKalphalacking both the E-F hand and the recoverin homology domain is constitutively active and is not further activated by any of the lipids tested or by calcium. However, a truncated form lacking only the recoverin homology domain is partially activated by either calcium or certain lipids. PMID- 15544348 TI - The alpha isoform of diacylglycerol kinase exhibits arachidonoyl specificity with alkylacylglycerol. AB - We compared the diacylglycerol kinase (DGK) catalyzed phosphorylation of 1-O hexanoyl-2-oleoylglycerol (HOG) with 1-O-hexanoyl-2-arachidonoylglycerol (HAG). We assayed the activity of DGKalpha and DGKzeta using a liposomal-based assay system. Liposomal assays show that the DGKalpha and, to a lesser extent, DGKzeta preferentially act on substrates containing an arachidonoyl group when this group is incorporated into alkylacylglycerols. The activity of DGKalpha was 82 times greater with HAG compared to HOG. DGKzeta is 10 times more active in catalyzing the phosphorylation of HAG compared to HOG. Although diacylglycerols were better substrates for both DGKalpha and DGKzeta than the alkylacylglycerols, no specificity was exhibited for arachidonoyl-containing diacylglycerols. However, this specificity for HAG over HOG is modulated by the phospholipid composition of the liposome. Addition of cholesterol and/or phosphatidylethanolamine partially reduces the substrate selectivity. We also analyzed the kinetic constants for the phosphorylation of both diacylglycerol and 1-alkyl-2-acylglycerol catalyzed by the alpha, epsilon, or zeta isoforms using a soluble Triton mixed micelle system. We found that all three isoforms of DGK can phosphorylate 1-alkyl-2-acylglycerols but generally at a lower rate than for the corresponding diacylglycerol. The specificity of DGKepsilon for diacylglycerols containing an arachidonoyl group was retained when the ester group in the C-1 position is replaced with an ether linkage. In contrast, DGKalpha and, to a lesser extent, DGKzeta had greater specificity for arachidonoyl-containing 1-alkyl-2-acylglycerols than for arachidonoyl-containing diacylglycerols. This demonstrates that the acyl chain specificity is affected by the structure of the lipid headgroup. PMID- 15544349 TI - A thermal unfolding study of plastocyanin from the thermophilic cyanobacterium Phormidium laminosum. AB - The thermal unfolding of the plastocyanin from Phormidium laminosum, a thermophilic cyanobacterium, is herein described. The main objective of this work is to identify structural factors responsible for the higher stability observed in proteins from thermophilic organisms. With the aid of fluorescence spectroscopy, EPR, and NMR, the factors influencing the unfolding process of the protein were investigated, and procedures for its study have been standardized. The different spectroscopic techniques used provided consistent results showing that the thermal unfolding of plastocyanin is irreversible under all the conditions investigated and that this irreversibility does not appear to be related to the presence of oxygen. The oxidized plastocyanin species has proven to be more stable than the reduced one, with respect to both the required temperature for protein unfolding (up to a 9 degrees C difference between the two forms) and the kinetics of the process. The behavior of this plastocyanin contrasts with that of other cupredoxins whose unfolding had previously been studied. The unfolding pH dependence and kinetic studies indicate a process with a tight control around the physiological pH in which plastocyanin plays its redox role and the protein's isoelectric point (5.2), suggesting a close compromise between function and stability. PMID- 15544350 TI - Subunit disassembly and unfolding kinetics of hemoglobin studied by time-resolved electrospray mass spectrometry. AB - We report the use of electrospray ionization (ESI) mass spectrometry (MS) in conjunction with online rapid mixing to monitor the kinetics of acid-induced ferrihemoglobin denaturation. Under equilibrium conditions, the hemoglobin mass spectrum is dominated by the intact heterotetramer. Dimeric and monomeric species are also observed at lower intensities. In addition, ionic signals corresponding to hexameric (tetramer-dimer) and octameric (tetramer x 2) hemoglobin species are observed. These complexes may represent weak solution-phase assemblies. The acid induced denaturation process was monitored for reaction time ranging from 9 ms to approximately 3 s. The data obtained were subjected to a global analysis procedure which simultaneously fit all kinetic (ESI-MS intensity vs time) profiles to multiexponential expressions. Results of the global analysis are consistent with the coexistence of two subpopulations of tetrameric hemoglobin which differ in their disassembly rates and ESI charge states. The higher-charge state tetramer ions preferentially dissociate via a rapid pathway (tau(1) = 51 ms), resulting in the transient formation of a heme-saturated dimer, holo-alpha globin, and a heme-deficient dimer. The latter is shown by MS/MS to be comprised of a heme-bound alpha-subunit complexed with an apo-beta-chain. The slow-decaying tetramer population, apparent at a slightly lower average charge state, breaks down into its monomeric constituents with no observable intermediate species (tau(2) = 390 ms). Surprisingly, unfolded apo-alpha-globin is formed more rapidly than unfolded apo-beta-globin. The appearance of the latter occurs with a relaxation time tau(3) of 1.2 s. It is postulated that accumulation of unfolded apo-beta-globin is delayed by transient population of an undetected unfolding intermediate. PMID- 15544351 TI - Constraints of the 9-methyl group binding pocket of the rhodopsin chromophore probed by 9-halogeno substitution. AB - Sterical constraints of the 9-methyl-binding pocket of the rhodopsin chromophore are probed using retinal analogues carrying substituents of increasing size at the 9 position (H, F, Cl, Br, CH(3), and I). The corresponding 11-Z retinals were employed to investigate formation of photosensitive pigment, and the primary photoproduct was identified by Fourier transform infrared difference spectroscopy. In addition, any effects of cumulative strain were studied by introduction of the 9-Z configuration and/or the alpha-retinal ring structure. Our results show that the 9-F analogue still can escape from the 9-methyl-binding pocket and that its photochemistry behaves very similar to the 9-demethyl analogue. The 9-Cl and 9-Br analogues behave very similar to the native 9-methyl pigments, but the 9-I retinal does not fit very well and shows poor pigment formation. This puts an upper limit on the radial dimension of the 9-methyl pocket at 0.45-0.50 nm. Introduction of the alpha-retinal ring constraint in the 11-Z series results in cumulative strain, because the 9-I and 9-Br derivatives cannot bind to generate a photopigment. The 9-Z configuration can partially compensate for the additional alpha-retinal strain. The corresponding 9-Br analogue does form a photopigment, and the other derivatives give increased photopigment yields compared to the corresponding 11-Z derivatives. In fact, 9-Z alpha-retinal would be an interesting candidate for retinal supplementation studies. Our data provide direct support for the concept that the 9-methyl group is an important determinant in ligand anchoring and activation of the protein and in general agree with a three-point interaction model involving the ring, 9 methyl group, and aldehyde function. PMID- 15544352 TI - Molecular mechanism of reverse cholesterol transport: reaction of pre-beta migrating high-density lipoprotein with plasma lecithin/cholesterol acyltransferase. AB - A 70-75 kDa high-density lipoprotein (HDL) particle with pre-beta-electrophoretic migration (pre-beta(1)-HDL) has been identified in several studies as an early acceptor of cell-derived cholesterol. However, the further metabolism of this complex has not been determined. Here we sought to identify the mechanism by which cell-derived cholesterol was esterified and converted to mature HDL as part of reverse cholesterol transport (RCT). Human plasma selectively immunodepleted of pre-beta(1)-HDL was used to study factors regulating pre-beta(1)-HDL production. A major role for phospholipid transfer protein (PLTP) in the recycling of pre-beta(1)-HDL was identified. Cholesterol binding, esterification by lecithin/cholesterol acyltransferase (LCAT) and transfer by cholesteryl ester transfer protein (CETP) were measured using (3)H-cholesterol-labeled cell monolayers. LCAT bound to (3)H-free cholesterol (FC)-labeled pre-beta(1)-HDL generated cholesteryl esters at a rate much greater than the rest of HDL. The cholesteryl ester produced in pre-beta(1)-HDL in turn became the preferred substrate of CETP. Selective LCAT-mediated reactivity with pre-beta(1)-HDL represents a novel mechanism increasing the efficiency of RCT. PMID- 15544353 TI - Treatment of cells with the angiogenic inhibitor fumagillin results in increased stability of eukaryotic initiation factor 2-associated glycoprotein, p67, and reduced phosphorylation of extracellular signal-regulated kinases. AB - Fumagillin, an angiogenic inhibitor, binds to methionine aminopeptidase 2, which is the same as eukaryotic initiation factor 2-associated glycoprotein, p67. p67 protects eIF2alpha from phosphorylation by its kinases. To understand the importance of fumagillin binding to p67, we measured the level of p67 in mouse C2C12 myoblasts treated with fumagillin. We show that fumagillin increases the stability of p67 by decreasing its turnover rate. The increased levels of p67 result in inhibition of phosphorylation of extracellular signal-regulated kinases 1 and 2 (ERKs 1 and 2). p67 binds to these ERKs, and the 108-480 amino acid segment is sufficient for this binding. p67's affinity to ERKs 1 and 2 also increases in fumagillin-treated myoblasts while its affinity for eIF2alpha remains unchanged. A mutant at the conserved amino acid residue D251A increases the phosphorylation of ERKs 1 and 2 without affecting the binding to p67, thus indicating the importance of this residue in the regulation of the phosphorylation of these ERKs. These results suggest that fumagillin increases the stability of p67 and its affinity to ERKs 1 and 2 and causes the inhibition of the phosphorylation of ERKs 1 and 2. PMID- 15544354 TI - Kinetic mechanism of blebbistatin inhibition of nonmuscle myosin IIb. AB - We examined the effect of blebbistatin on the kinetic properties of nonmuscle myosin IIB subfragment 1 (NMIIB S1). Blebbistatin is a small molecule that affects cell blebbing during the process of cell division, which has been shown to decrease the myosin ATPase activity of a number of myosins [Straight et al. (2003) Science 299, 1743-1747]. The steady-state actin-activated ATPase activity of NMIIB S1 was decreased approximately 90% at 40 microM actin in the presence of blebbistatin. Stopped-flow techniques were employed to elucidate the effect of blebbistatin on the various steps of the NMIIB S1 cross-bridge cycle. Blebbistatin did not affect ATP binding and hydrolysis. Binding to actin in the presence of ADP (0.57 +/-0.08 microM(-1) s(-1)) was reduced slightly in the presence of blebbistatin (0.38 +/- 0.03 microM(-1) s(-1)), while mantADP dissociation from acto-NMIIB S1 was reduced (approximately 30%). P(i) release was blocked in the presence of blebbistatin. Accordingly, the apparent affinity of NMIIB S1 for actin in the presence of ATP was greatly reduced. Based on the above data, we surmise that blebbistatin inhibits the ATPase activity of NMIIB S1 primarily by blocking entry into the strong binding state; secondarily, it reduces the rate of ADP release. These effects are likely mediated by binding of blebbistatin within the myosin cleft that progressively closes in forming the acto-myosin rigor state. PMID- 15544355 TI - The rate of P-glycoprotein activation depends on the metabolic state of the cell. AB - P-glycoprotein ATPase activity has been studied almost exclusively by measuring inorganic phosphate release from inside-out cellular vesicles. We have recently proposed a new method based on measurements of the extracellular acidification rate (ECAR) of living cells with a Cytosensor microphysiometer. This method allows for systematic investigation of the various factors influencing P glycoprotein activation in living cells. Basal metabolic rates or ECARs of different MDR1-transfected cell lines were compared with those of the Mdr1a(-/ )1b(-/-) knockout, MRP1-transfected, and corresponding wild-type cell lines. Basal ECARs of all cells were on the order of 10(7) protons/cell/s, whereby those of genetically modified cells were on average (over all cell lines) slightly lower than those of wild-type cells. The expression level of P-glycoprotein in MDR1-transfected cells had no influence on basal ECARs. Verapamil-induced ECARs were specific for MDR1-transfected cells and increased with the expression level of P-glycoprotein. Moreover, ECARs were dependent on the metabolic state of the cell and were (2.8 +/- 1.2) x 10(6) and (8.0 +/- 1.5) x 10(6) protons/cell/s in glucose-deficient and glucose-fed NIH-MDR-G185 cells, respectively, after verapamil (10 muM) stimulation. The ECARs were practically identical to the rates of lactate extrusion and thus reflect the rates of ATP synthesis via glycolysis. Taking into account the number of P-glycoprotein molecules per cell, the rate of ATP hydrolysis in inside-out vesicles of the same cells was determined as (9.2 +/ 1.5) x 10(6) phosphates/cell/s, in good agreement with the rate of ATP synthesized in glucose-fed cells. The energy required for P-glycoprotein activation relative to the basal metabolic energy was twice as large in glucose deficient as in glucose-fed cells, suggesting cellular protection by P glycoprotein even under conditions of starvation. PMID- 15544356 TI - Real-time measurement of in vitro peptide binding to soluble HLA-A*0201 by fluorescence polarization. AB - Measuring the interaction of class I human leukocyte antigens (HLA) and their peptide epitopes acts as a guide for the development of vaccines, diagnostics, and immune-based therapies. Here, we report the development of a sensitive biochemical assay that relies upon fluorescence polarization to indicate peptide interactions with recombinant soluble HLA proteins. It is a cell- and radioisotope-free assay that has the advantage of allowing the direct, real-time measurement of the ratio between free and bound peptide ligand in solution without separation steps. Peptide/HLA assay parameters were established using several HLA A*0201-specific fluorescein isothiocyanate-labeled peptides. Optimal loading of synthetic peptides into fully assembled soluble HLA-A*0201 complexes was enabled by thermal destabilization at 53 degrees C for 15 min, demonstrating that efficient peptide exchange does not require the removal of endogenous peptides from the reaction environment. An optimal ratio of three beta-2 microglobulin molecules per single HLA heavy chain was determined to maximize peptide binding. Kinetic binding studies indicate that soluble HLA-A*0201/peptide interactions are characterized by a range of moderate k(on) values (1 x 10(4) to 8.7 x 10(4) M(-1) s(-1)) and slow k(off) values (1.9 x 10(-4) to 4.3 x 10(-4) s( 1)), consistent with parameters for native HLA molecules. Testing of the A*0201 specific peptides with 48 additional class I molecules demonstrates that the unique peptide binding behavior of individual HLA molecules is maintained in the assay. This assay therefore represents a versatile tool for characterizing the binding of peptide epitopes during the development of class I HLA-based vaccines and immune therapies. PMID- 15544357 TI - Full or partial substitution of the reactive center loop of alpha-1-proteinase inhibitor by that of heparin cofactor II: P1 Arg is required for maximal thrombin inhibition. AB - The abundant plasma protein alpha(1)-proteinase inhibitor (alpha(1)-PI) physiologically inhibits neutrophil elastase (NE) and factor XIa and belongs to the serine protease inhibitor (serpin) protein superfamily. Inhibitory serpins possess a surface peptide domain called the reactive center loop (RCL), which contains the P1-P1' scissile peptide bond. Conversion of this bond in alpha(1)-PI from Met-Ser to Arg-Ser in alpha(1)-PI Pittsburgh (M358R) redirects alpha(1)-PI from inhibiting NE to inhibiting thrombin (IIa), activated protein C (APC), and other proteases. In contrast to either the wild-type or M358R alpha(1)-PI, heparin cofactor II (HCII) is a IIa-specific inhibitor with an atypical Leu-Ser reactive center. We examined the effects of replacement of all or part of the RCL of alpha(1)-PI with the corresponding parts of the HCII RCL on the activity and specificity of the resulting chimeric inhibitors. A series of 12 N-terminally His tagged alpha(1)-PI proteins differing only in their RCL residues were expressed as soluble proteins in Escherichia coli. Substitution of the P16-P3' loop of alpha(1)-PI with that of HCII increased the low intrinsic antithrombin activity of alpha(1)-PI to near that of heparin-free HCII, while analogous substitution of the P2'-P3' dipeptide surpassed this level. However, gel-based complexing and quantitative kinetic assays showed that all mutant proteins inhibited thrombin at less than 2% of the rate of alpha(1)-PI (M358R) unless the P1 residue was also mutated to Arg. An alpha(1)-PI (P16-P3' HCII/M358R) variant was only 3-fold less active than M358R against IIa but 70-fold less active against APC. The reduction in anti-APC activity is desired in an antithrombotic agent, but the improvement in inhibitory profile came at the cost of a 3.5-fold increase in the stoichiometry of inhibition. Our results suggest that, while P1 Arg is essential for maximal antithrombin activity in engineered alpha(1)-PI proteins, substitution of the corresponding HCII residues can enhance thrombin specificity. PMID- 15544358 TI - In situ characterization of aluminum-containing mineral-microorganism aqueous suspensions using scanning transmission X-ray microscopy. AB - In situ characterization of colloidal particles under hydrous conditions is one of the key requirements for understanding their state of aggregation and impact on the transport of pollutants in aqueous environments. Scanning transmission X ray microscopy (STXM) is one of the few techniques that can satisfy this need by providing element- and chemical-state-specific 2-D maps at a spatial resolution better than 50 nm using soft X-rays from synchrotron radiation wiggler or undulator sources tuned to the absorption edges of different elements. X-ray absorption near-edge structure (XANES) spectra can also be collected simultaneously at a similar spatial resolution and can provide phase identification in many cases. In this study, we report STXM images and XANES spectroscopy measurements at or above the Al K-edge (E = 1559.6 eV) of various Al containing minerals and synthetic oxides [alpha-Al2O3 (corundum), gamma-Al2O3, gamma-AlOOH (boehmite), alpha-Al(OH)3 (bayerite), KAl2(AlSi3O10)(OH)2 (muscovite), (Al,Mg)8(Si4O10)4(OH)8.nH2O (montmorillonite), and Mg6Al2(OH)16CO3.4H2O (hydrotalcite)] and demonstrate the capability of this spectromicroscopic tool to identify different Al-containing mineral colloids in multiphase mixtures in aqueous solution. We also demonstrate that STXM imaging at or above the C K-edge (E = 284.2 eV) and Al K-edge can provide unique information on the interactions between bacteria and Al-containing nanoparticles in aqueous suspensions. STXM images of a mixture of Caulobacter crescentus and montmorillonite and corundum particles just above the C and Al K-edges show that the mineral particles and bacteria are closely associated in aggregates, which is likely due to the binding of bacteria to clay and corundum particles by extracellular polysaccharides. PMID- 15544359 TI - In situ measurements of nanotube dimensions in suspensions by depolarized dynamic light scattering. AB - We show that the dimensions of carbon nanotubes (CNTs) in suspension can be characterized by depolarized dynamic light scattering. Taking advantages of this in situ technique, we investigate in detail the influence of sonication procedures on the length and diameter of CNTs in surfactant solutions. Sonication power is shown to be particularly efficient at unbundling nanotubes, whereas a long sonication time at low power can be sufficient to cut the bundles with limited unbundling. We finally demonstrate the influence of CNT dimensions on the electrical properties of CNT fibers. Slightly varying the sonication conditions, and thereby the suspended nanotube dimensions, can affect the fibers conductivity by almost 2 orders of magnitude. PMID- 15544360 TI - Foam superstabilization by polymer microrods. AB - Few foam systems stabilized by solid particles have been reported, and usually the particles have been used in combination with surfactants. We report how foams can be stabilized solely with a new class of anisotropic particles, hydrophobic polymer microrods of diameter less than 1 mum and length of a few tens of micrometers. The obtained foams were extraordinary stable, retaining a constant volume over many days and even surviving drying of most of the free liquid. The bubbles in these foams were sterically stabilized by dense thick "hairy" layers. The rigid intertwined protective shells around the bubbles did not allow the formation of thin films between them. The lifetime of these foams was orders of magnitude longer than the ones stabilized with typical foaming surfactants such as sodium dodecyl sulfate. The addition of sodium dodecyl sulfate led to hydrophilization of the microrods and suppressed the superstabilization effect. Thus, common foaming agents effectively act as defoamers for the ultrastable foams stabilized by microrods. PMID- 15544361 TI - Preparation and photoinduced patterning of azidoformate-terminated self-assembled monolayers. AB - This paper presents a new method for the patterning of self-assembled monolayers (SAMs) using UV light. Azidoformate-terminated SAMs starting from 18-acetoxy octadecyltrichlorosilane SAMs on silicon, prepared for the first time, are electrophilic and photosensitive, and can be patterned by UV irradiation through a mask. The resulting structured surfaces are still electrophilic and can be reacted with nucleophilic functions, for example, primary amines. PMID- 15544362 TI - Synthesis and characterization of phosphido-monolayer-protected gold nanoclusters. AB - Gold-phosphido-monolayer-protected clusters (MPCs) of 1-2-nm diameter, Au(x)(PR2)y, analogues of the well-known thiolate materials Au(x)(SR)y, were prepared by NaBH4 reduction of a mixture of HAuCl4.3H2O and a secondary phosphine PHR2 in tetrahydrofuran/water. In comparison to the Au-thiolate MPCs, fewer of the larger phosphido groups are required to cover the surface, and the Au-P bond is not cleaved as readily in reactions with small molecules as is its Au-S counterpart. 31P NMR spectroscopy provides a direct method to study cluster formation and the interaction of the phosphido ligand with the gold surface. PMID- 15544363 TI - Dynamic light scattering studies of poly(ethylene oxide) adsorbed on Laponite: layer conformation and its effect on particle stability. AB - Dynamic light scattering has been used to determine the hydrodynamic thickness of poly(ethylene oxide) (PEO) adsorbed on synthetic anisotropic clay particles (Laponite) as a function of molecular weight. The layer thicknesses, and their increase with molecular weight, indicate that the conformation of the adsorbed layer is very compact and is much smaller than those normally observed for polymer adsorption on flat interfaces. The aggregation kinetics of the polymer coated particles in 5 mM NaCl was analyzed in a quantitative manner, revealing that the potential barrier to aggregation is strongly enhanced when polymer is present. PMID- 15544364 TI - Preparation and surface properties of low-density gels synthesized using prepolymerized silica precursors. AB - Properties of silica xerogels and aerogels synthesized using a number of prepolymerized silica precursors were probed by 29Si magic-angle spinning (MAS) NMR spectroscopy, the small-angle X-ray scattering (SAXS) method, the nitrogen adsorption method, and transmission electron microscopy (TEM) to show that xerogels with attractive textural properties can easily be prepared using this type of precursors and the conventional one-step, base procedure. Pore sizes and overall pore volumes in these materials can be notably larger than those in the corresponding materials synthesized using tetraethoxysilane. This positive effect stems from the stronger structure of the polymeric network due to a higher degree of silica condensation on one side and a larger thickness of polymeric chains on the other. The thorough investigations of the fine silica structure demonstrate, however, that the relationship between the microstructure of the silica precursor and the micro- and macrostructures of dry gels is complex and the use of more condensed precursors favors, but does not necessarily ensure, more porous dry materials, under the same reaction conditions. Ethyl silicate 40 may be recommended as a low-cost precursor suitable for applications in this situation. PMID- 15544365 TI - Influence of free protein on flocculation stability of beta-lactoglobulin stabilized oil-in-water emulsions at neutral pH and ambient temperature. AB - The influence of protein concentration and order of addition relative to homogenization (before or after) on the extent of droplet flocculation in oil-in water emulsions stabilized by a globular protein was examined using laser diffraction. n-Hexadecane (10 wt%) oil-in-water emulsions (pH 7, 150 mM NaCl) stabilized by beta-lactoglobulin (beta-Lg) were prepared by three methods: (1) 4 mg/mL beta-Lg added before homogenization; (2)10 mg/mL beta-Lg added before homogenization; (3) 4 mg/mL beta-Lg added before homogenization and 6 mg/mL beta Lg added after homogenization. Emulsion 1 contained little nonadsorbed protein (<3%) and underwent extremely rapid and extensive droplet flocculation immediately after homogenization. Emulsion 2 contained a significant fraction of nonadsorbed beta-Lg and exhibited relatively slow droplet flocculation for some hours after homogenization. Measurements on Emulsion 3 showed that the extremely rapid particle growth observed in Emulsion 1 could be arrested by adding native beta-Lg immediately after homogenization. The extent of particle growth in the three types of emulsions was highly dependent on the time that the salt was added to the emulsions, i.e., after 0 or 24 h aging. We postulate that the observed differences are due to changes in droplet surface hydrophobicity caused by differences in the packing or conformation of adsorbed proteins. Our data suggest that history effects have a strong influence on the flocculation stability of protein-stabilized emulsions, which has important implications for the formulation and production of protein stabilized oil-in-water emulsions. PMID- 15544366 TI - Interactions of charged porphyrins with nonionic triblock copolymer hosts in aqueous solutions. AB - The extent and locus of solubilization of guest and self-assembling surfactant host molecules in aqueous solutions are influenced by a variety of hydrophobic and hydrophilic interactions, as well as by more specific interactions between the various species present. By using a combination of two-dimensional heteronuclear 13C[1H] NMR correlation experiments with pulsed-gradient NMR diffusion and proton cross-relaxation measurements, the locations and distributions of porphyrin guest molecules have been established unambiguously with respect to the hydrophobic and hydrophilic moieties of a triblock copolymer species in solution. The interactions of tetra(4-sulfonatophenyl)porphyrin with the poly(propylene oxide) (PPO) and the poly(ethylene oxide) (PEO) segments of amphiphilic PEO-PPO-PEO triblock copolymer species have been measured as functions of solution conditions, including temperature and pH. The porphyrin/PEO PPO-PEO interactions are established to be selective and adjustable according to the different temperature-dependent hydrophilicities or hydrophobicities of the PEO and PPO triblock copolymer components. Furthermore, such interactions influence the self-assembly properties of the block-copolymer amphiphiles in solution by stabilizing molecular porphyrin/PEO-PPO-PEO complexes well above the critical micellization temperature of the triblock copolymer species under otherwise identical conditions. PMID- 15544367 TI - Nonpolymeric hydrogelator derived from N-(4-pyridyl)isonicotinamide. AB - A series of pyridyl amides derived from isonicotinic acid, nicotinic acid, and benzoic acid have been synthesized. Only N-(4-pyridyl)isonicotinamide 1 is found to be an efficient hydrogelator with a minimum gelator concentration of 0.37 wt %. A wide range of concentrations (0.37-20 wt %) could be used to form hydrogels. The other amides, namely, N-(3-pyridyl)isonicotinamide 2, N-(2 pyridyl)isonicotinamide 3, N-(phenyl)isonicotinamide 4, N-(4-pyridyl)nicotinamide 5, N-(3-pyridyl)nicotinamide 6, and N-(4-pyridyl)benzamide 7, did not show any gelation properties. Fourier transform infrared spectroscopy, variable temperature 1H NMR, single-crystal diffraction and X-ray powder diffraction (XRPD), and scanning electron microscopy have been used to characterize the gel. Single-crystal diffraction and XRPD studies indicate that the morph responsible for gel formation is different from that in its bulk solid and xerogel. PMID- 15544368 TI - Stability, interaction, size, and microenvironmental properties of mixed micelles of decanoyl-N-methylglucamide and sodium dodecyl sulfate. AB - The mixed micellization between the nonionic surfactant decanoyl-N methylglucamide (MEGA-10) and the common sodium dodecyl sulfate (SDS) in aqueous solutions of 0.1 M NaCl was investigated by the fluorescence probe method. The critical micelle concentrations were determined by the pyrene 1:3 ratio method. The experimental data are discussed in light of two mixing thermodynamic models within the framework of the pseudophase separation model, including the conventional regular solution theory and a recent treatment proposed by Maeda (J. Phys. Chem. B 2004, 108, 6043). This last approach provides a more appropriate description of the mixed system, particularly in two aspects: the nature of the interactions responsible for the stability of the mixed micelle and the behavior of the excess free energy per monomer of the system. By using the static quenching method, the mean micellar aggregation numbers of mixed micelles in the whole range of compositions were obtained. It was found that the micellar aggregation number initially increases with the content of the ionic component, then remains roughly constant, and, finally, decreases slightly for high content of this component. This behavior was analyzed taking into account the effects produced by the presence of the charged headgroups of sodium dodecyl sulfate, as this component increases its participation in the mixed micelle. The micropolarity of the mixed micelles was studied by the pyrene 1:3 ratio index. It was observed that the increasing participation of the ionic component induces the formation of micelles with a more dehydrated structure. Data of micellar microviscosity were obtained by using different methods, including fluorescence intensity measurements of Auramine O and steady-state fluorescence anisotropy of rhodamine B and diphenylbutadiene. The results obtained from these experiments are in good agreement and suggest the formation of mixed micelles with a less ordered structure as the content of SDS increases. PMID- 15544369 TI - Shamrock surfactants: synthesis and characterization. AB - Two types of a new class of surfactants with three headgroups, which possess the general structure 1, have been prepared. Within structure 1, a central headgroup is connected to two flanking headgroups by hydrocarbon chains. The term "shamrock" is used to describe surfactants of structure 1, denoting their triple headed character and reflecting the fact that shamrocks have leaflets in groups of three. The major lipophilic character of shamrock surfactants is provided by the two hydrocarbon chains linking the three headgroups and not by long-chain alkyl groups appended to the linking hydrocarbon chains or the headgroups. The new surfactants are 2a (2,2,15,15,28,28-hexamethyl-2,15,28-triazonianonacosane triiodide), 2b (2,2,15,15,28,28-hexamethyl-2,15,28-triazonianonacosane trichloride), 3a (O,O'-di-[10-(N,N,N-tripropylammonio)decyl]phosphorodithioate bromide), and 3b (O,O'-di-[10-(N,N,N-tributylammonio)decyl]phosphorodithioate bromide). Compound 14 (2,2,9,9,16,16-hexamethyl-2,9,16-triazoniaheptadecane triiodide) was prepared for comparison with 2a. Surfactants 2 and 3 were characterized in water by measurement of their Krafft temperatures and critical aggregation concentrations, and their aggregates were studied by 1H NMR spectroscopy, dynamic laser light scattering, and phase-contrast optical microscopy. Aqueous 2b was also studied by cryo-etch high-resolution scanning electron microscopy, which revealed irregularly shaped cells containing a complex matrix of surfactant. Coacervates were observed by optical microscopy upon the hydration of 2 and 3. PMID- 15544370 TI - Microemulsion efficiency boosting and the complementary effect. 1. Structural properties. AB - Amphiphilic diblock copolymers added to microemulsions proved to enhance the efficiency of surfactants dramatically. The complementary effect of homopolymers is considered in the current work. A possible application of the homopolymer addition could be the viscosity tuning of the microemulsion without changing the considered bicontinuous phase. Furthermore, (homo)polymers are added for many other reasons in technical applications. A theory by Eisenriegler predicts a decreased efficiency when homopolymers are added. In further experiments, the simultaneous addition of homopolymers and diblock copolymers probes whether the two opposite effects superpose and allow for a compensation. Then, efficiency and viscosity are adjustable independently. Experimentally, phase diagrams are investigated and the microscopic structure is measured by small-angle neutron scattering. Within the presented models, both experimental methods are compared and discussed on the basis of the surfactant membrane bending moduli. The homopolymer effect is about 7 times larger than that theoretically predicted, and the superposition of the two polymer effects allows for a compensation with an optionally tunable viscosity. PMID- 15544371 TI - Petunia-shaped superstructures of CaCO3 aggregates modulated by modified chitosan. AB - This paper presents the crystallization behavior of calcium carbonate at the air/liquid interface of aqueous systems of carboxymethyl chitosan (CMCS) using the Kitano method. Although the synthesized CMCS (Mw approximately 100,000) with 1.57 degree of carboxymethyl substitution shows no surface activity, it controls the crystallization of calcium carbonate to form a petunia-shaped superstructure. The shuttlecock-like head of this superstructure strongly supports Colfen's opinion (Rudloff, J.; Colfen, H. Langmuir 2004, 20, 991-996) for the existence of a gas template of CO2 bubbles temporarily captured by polymer molecules, while formation of the stem of this superstructure is provisionally attributed to the presence of the strong electrostatic interactions between calcium ions and the carboxylate groups. The CaCO3 superstructure and its morphology depend not only on the polymer concentration but also on the combined number of calcium ions per CMCS molecule. These results imply that this simple and versatile method expands the morphological investigation of mineralization processes. PMID- 15544372 TI - Polymer bilayer formation due to specific interactions between beta-cyclodextrin and adamantane: a surface force study. AB - The purposes of this study are to utilize the interactions between an adamantane end-capped poly(ethylene oxide) (PEO) and a cationic polymer of beta-cyclodextrin to build polymer bilayers on negatively charged surfaces, and to investigate the interactions between such layers. The association of this system in solution has been studied by rheology, light scattering, and fluorescence measurements. It was found that the adamantane-terminated PEO (PEO-Ad) mixed with the beta cyclodextrin polymer gives complexes where the interpolymer links are formed by specific inclusion of the adamantane groups in the beta-cyclodextrin cavities. This results in a higher viscosity of the solution and growth of intermolecular clusters. The interactions between surfaces coated with a cationized beta cyclodextrin polymer across a water solution containing PEO-Ad polymers were studied by employing the interferometric surface force apparatus (SFA). In the first step, the interaction between mica surfaces coated with the cationized beta cyclodextrin polymer in pure water was investigated. It was found that the beta cyclodextrin polymer adsorbs onto mica and almost neutralizes the surface charge. The adsorbed layers of the beta-cyclodextrin polymer are rather compact, with a layer thickness of about 60 A (30 A per surface). Upon separation, a very weak attractive force is observed. The beta-cyclodextrin solution was then diluted by pure water by a factor of 3000 and a PEO-Ad polymer was introduced into the solution. Two different architectures of the PEO-Ad polymer were investigated: a four-arm structure and a linear structure. After the adsorption of the PEO polymer onto the beta-cyclodextrin layer reached equilibrium, the forces were measured again. It was found that the weak repulsive long-range force had disappeared and an attractive force caused the surfaces to jump into contact, and that the compressed layer thickness had increased. The attractive force is interpreted as being due to a specific recognition between the hydrophobic adamantane groups on the PEO-Ad polymer and the hydrophobic cavity in the beta cyclodextrin molecules. Furthermore, the attractive force observed on separation has increased significantly, which is a further indication of a specific interaction between the beta-cyclodextrin polymer and the adamantane groups. PMID- 15544373 TI - Surface self-assembly of fluorosurfactants during film formation of MMA/nBA colloidal dispersions. AB - These studies focus on the behavior of fluorosurfactants (FS) containing hydrophobic and ionic entities in the presence of methyl methacrylate/n-butyl acrylate (MMA/nBA) colloidal dispersions stabilized by sodium dodecyl sulfate (SDS). The presence of FS significantly not only alters the mobility of SDS in MMA/nBA films, but their hydrophobic and ionic nature results in self-assembly near the film-air (F-A) interface leading to different surface morphologies. Spherical islands and rodlike morphologies are formed which diminish the kinetic coefficient of friction of films by at least 3 orders of magnitude, and the presence of dual hydrophobic tails and an anionic head appears to have the largest effect on the surface friction. Using internal reflection IR imaging, these studies show that structural and chemical features of FS are directly related to their ability to migrate to the F-A interface and self-assemble to form specific morphological features. While the anionic nature of FS allows for SDS migration to the F-A interface and the formation of stable domains across the surface, intermolecular cohesion of nonionic FS allows for the formation of rodlike structures due to inability to form mixed micelles with SDS. These studies also establish the relationship between surface morphologies, kinetic coefficient of friction, and structural features of surfactants in the complex environments. PMID- 15544374 TI - Immobilization of the enzyme beta-lactamase on biotin-derivatized poly(L-lysine) g-poly(ethylene glycol)-coated sensor chips: a study on oriented attachment and surface activity by enzyme kinetics and in situ optical sensing. AB - Understanding the conformation, orientation, and specific activity of proteins bound to surfaces is crucial for the development and optimization of highly specific and sensitive biosensors. In this study, the very efficient enzyme beta lactamase is used as a model protein. The wild-type form was genetically engineered by site-directed mutagenesis to introduce single cysteine residues on the surface of the enzyme. The cysteine thiol group is subsequently biotinylated with a dithiothreitol (DTT)-cleavable biotinylation reagent. beta-Lactamase is then immobilized site-specifically via the biotin group on neutral avidin-covered surfaces with the aim to control the orientation of the enzyme molecule at the surface and study its effect on enzymatic activity using Nitrocefin as the substrate. The DTT-cleavable spacer allows the release of the specifically bound enzyme from the surface. Immobilization of the enzyme is performed on a monolayer of the polycationic, biotinylated polymer PLL-g-PEG/PEG-biotin assembled on niobium oxide (Nb2O5) surfaces via neutral avidin as the docking site. Two different assembly protocols, the sequential adsorption of avidin and biotinylated beta-lactamase and the immobilization of preformed complexes of beta lactamase and avidin, are compared in terms of immobilization efficiency. In situ optical waveguide lightmode spectroscopy and colorimetric analysis of enzymatic activity were used to distinguish between specific and unspecific enzyme adsorption, to sense quantitatively the amount of immobilized enzyme, and to determine Michaelis-Menten kinetics. All tested enzyme variants turned out to be active upon immobilization at the polymeric surface. However, the efficiency of immobilized enzymes relative to the soluble enzymes was reduced about sevenfold, mainly because of impaired substrate (Nitrocefin) diffusion or restricted accessibility of the active site. No significant effect of different enzyme orientations could be detected, probably because the enzymes were attached to the surface through long, flexible PEG chain linkers. PMID- 15544375 TI - Time evolution studies of the H2O/quartz interface using sum frequency generation, atomic force microscopy, and molecular dynamics. AB - Many interfacial studies on solid surfaces, for example, quartz/water, assume that a standard cleaning procedure regenerates the surface reproducibly. In the reported work, the results of two surface specific techniques, sum frequency generation (SFG) spectroscopy and atomic force microscopy, show that the effects of prolonged exposure to Nanopure water and to pH 10 NaOH are distinctly different. In conjunction with the experimental data, molecular mechanics is used to correlate the SFG spectral frequencies to the hydrogen stretching vibrations of the surface-bound water molecules. It is found that after 17 days of soaking in water, water molecules penetrate into the SiO2 matrix to produce a swollen and amorphous layer; it is likely that broken Si-O bonds from the polishing process serve as nucleation sites for hydration and swelling. Disorder introduced in the interfacial water layer is detected by the rising intensity of the weakly hydrogen-bonded SFG peak at 3450 cm(-1). Dominance of the 3450 cm(-1) is absent in a pH 10, NaOH-soaked quartz disk, indicating that the strong hydrogen-bonded network in water remains intact. PMID- 15544376 TI - Plasma surface modification of organic materials: comparison between polyethylene films and octadecyltrichlorosilane self-assembled monolayers. AB - Low-pressure low-frequency NH3 plasmas have been used for the surface modification of bulk polyethylene films and of octadecyltrichlorosilane (OTS) self-assembled monolayers deposited on oxidized silicon wafers. The incorporation of nitrogen-containing groups by the plasma treatment has been followed by contact angle measurements and by X-ray photoelectron spectroscopy. The surface degradation of the OTS monolayers due to plasma etching has been measured separately by optical ellipsometry with subnanometric accuracy. Our data show clear evidence for the existence of an optimum treatment time, yielding a high density of NH2 functional groups without significant variation of the structural features of the organic material. Self-assembled monolayers appear as excellent model systems to characterize the effects of plasma discharges on polyolefins. In particular, they allow testing the influence of molecular orientation, packing density, and crystallinity on the final results. PMID- 15544377 TI - Infrared study of CO adsorbed on Pd/Al2O3-ZrO2. Effect of zirconia added by impregnation. AB - Characterization of palladium catalysts, supported on alumina and alumina modified by zirconia added by impregnation, was performed by CO adsorption from 143 to 298 K and monitored by infrared spectroscopy. It was found that the population of the Al3+ octahedral sites in the alumina decreased by the addition of zirconia. In contrast to the case of the pure alumina support, where stabilization of Pd+ was observed, Pd2+ was formed preferentially on samples where zirconia was added, and higher crystallinity in the metallic palladium was observed. Studies of CO adsorption at low temperatures (143 K) gave a better description of the surface species, since at higher temperatures (298 K) the reaction of the CO with some of the palladium oxide particles led to the partial reduction of the latter. PMID- 15544378 TI - Control of surface properties using fluorinated polymer brushes produced by surface-initiated controlled radical polymerization. AB - Surface-grafted styrene-based homopolymer and diblock copolymer brushes bearing semifluorinated alkyl side groups were synthesized by nitroxide-mediated controlled radical polymerization on planar silicon oxide surfaces. The polymer brushes were characterized by X-ray photoelectron spectroscopy (XPS), near-edge X ray absorption fine structure (NEXAFS), and time-dependent water contact angle measurements. Angle-resolved XPS studies and water contact angle measurements showed that, in the case of the diblock copolymer brushes, the second block to be added was always exposed at the polymer-air interface regardless of its surface energy. Values of z*/Rg were estimated based on the radius of gyration, Rg, of the grafted homopolymer or block copolymer chains for the grafted brushes and thickness of the brush, z*. The fact that z*/Rg > 1 suggests that all these brushes are stretched. These results support the idea that after grafting the first block onto the surface the nitroxide-end capped polymer chains were able to polymerize the second block in a "living" fashion and the stretched brush so formed was dense enough that the outermost block in all cases completely covers the surface. NEXAFS analysis showed a relationship between the surface orientation of the fluorinated side chains and brush thickness with thicker brushes having more oriented side chains. Time-dependent water contact angle measurements revealed that the orientation of the side chains of the brush improved the surface stability toward reconstruction upon prolonged exposure to water. PMID- 15544379 TI - Dansylated aminopropyl controlled pore glass: a model for silica-liquid solvation. AB - We have prepared a series of aminopropyl controlled pore glass (CPG) particles that have been labeled with a solvatochromic fluorescent probe molecule (dansyl). We report on the behavior of the attached dansyl reporter as a function of dansyl to-amine molar ratio (i.e., dansyl loading), solvent dipolarity, and surface residue end capping. In these experiments, we systematically adjust the dansyl loading by 10(5); a range much larger than ever explored. The dansylated CPG particles were also end capped with trimethylchlorosilane to derivatize most of the residual silanol and/or aminopropyl groups. The attached dansyl molecules can be surrounded by other dansyl molecules; they can be distributed within an ensemble of sites with differing physicochemical properties, and/or they can be distributed in sites that are restrictive to dansyl motion and/or solvent inaccessible. At high dansyl loadings, the majority of the dansyl groups are solvated by other dansyl moieties and solvent does not significantly alter the local microenvironment surrounding the average dansyl molecule (i.e., the cybotactic region) to any significant level. At intermediate dansyl loadings, the average distance between the dansyl groups increases and solvent is able to access/solvate/wet the dansyl groups and alter their cybotactic region to a greater extent. At the lowest dansyl loadings studied, the results suggest that these dansyl moieties are localized within solvent inaccessible/restrictive SiO2 sites (e.g., small pores). PMID- 15544380 TI - Characterization of polyelectrolyte multilayers by the streaming potential method. AB - Polyelectrolyte multilayer adsorption on mica was studied by the streaming potential method in the parallel-plate channel setup. The technique was calibrated by performing model measurements of streaming potential by using monodisperse latex particles. Two types of polyelectrolytes were used in our studies: poly(allylamine) hydrochloride (PAH), of a cationic type, and poly(sodium 4-styrenesulfonate) (PSS) of an anionic type, both having molecular weight of 70,000. The bulk characteristics of polymers were determined by measuring the specific density, diffusion coefficient for various ionic strengths, and zeta potential. These measurements as well as molecular dynamic simulations of chain shape and configurations suggested that the molecules assume an extended, wormlike shape in the bulk. Accordingly, the diffusion coefficient was interpreted in terms of a simple hydrodynamic model pertinent to flexible rods. These data allowed a proper interpretation of polyelectrolyte multilayer adsorption from NaCl solutions of various concentrations or from 10(-3) M Tris buffer. After completing a bilayer, periodic variations in the apparent zeta potential between positive and negative values were observed for multilayers terminated by PAH and PSS, respectively. These limiting zeta potential values correlated quite well with the zeta potential of the polymers in the bulk. The stability of polyelectrolyte films against prolonged washing (reaching 26 h) also was determined using the streaming potential method. It was demonstrated that the PSS layer was considerably more resistant to washing, compared to the PAH layer. It was concluded that the experimental data were consistent with the model postulating particle-like adsorption of polyelectrolytes with little chain interpenetration. It also was concluded that due to high sensitivity, the electrokinetic method applied can be effectively used for quantitative studies of polyelectrolyte adsorption, desorption, and reconformation. PMID- 15544381 TI - Interaction of sodium polyacrylate adsorbed on TiO2 with cationic and anionic surfactants. AB - Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) was used to identify the structures formed during the adsorption of sodium polyacrylate (NaPA) on charged TiO2 particles and to determine the subsequent interaction of the adsorbed polymer structure with cationic and anionic surfactants. The nature of the polymer structure was deduced from the adsorbed amount in tandem with the information obtained from monitoring the change in the relative intensity of the COO- and COOH infrared bands. In particular, it is found that the relative number of COO- and COOH groups on the polymer backbone for the adsorbed state differs from that of the same polymer in solution. This difference is due to a shift in the population of COO-/COOH groups on the polymer backbone that arises when the COO- groups bind to positively charged sites on the surface. A change in the number COO-/COOH groups on the polymer is thus related to a change in the bound fraction of polymer. It is shown that the initial NaPA approaching the bare surface adopts a flat conformation with high bound fraction. Once the bare sites on the surface are covered, the accommodation of additional polymer on the surface requires the existing adsorbed layer to adopt a conformation with a lower bound fraction. When the adsorbed NaPA is probed with a solution containing the anionic surfactant sodium dodecyl sulfate (SDS), the SDS competes for surface sites and displaces some of the bound NaPA segments from the surface, giving rise to an polymer layer adsorbed with an even lower bound fraction. In contrast, addition of a solution containing the cationic surfactant cetyltrimethylammonium bromide (CTAB) results in the binding of the surfactant directly to the free COO- sites on the adsorbed polymer backbone. Confirmation of a direct interaction of the CTAB headgroup with the free COO- groups of the polymer is provided by intensity changes in the headgroup IR bands of the CTAB. PMID- 15544382 TI - n-Decyl-glucopyranoside and n-decyl-maltopyranoside gibbs monolayers. Phase changes in the dilute liquid-expanded range. AB - Surface tension isotherms were recorded for n-decyl-beta-d-glucopyranoside (Glu) and n-decyl-beta-D-maltopyranoside (Mal) solutions at temperatures of 8, 22, and 29 degrees C. Comparison was made with isotherms of n-decyl-beta-D thiomaltopyranoside (S-Mal) at 22 degrees C. In addition to the transition from the gaseous to the liquid-expanded (LE) state, a second transition was observed in the early stages of the LE regime for Glu, Mal, and S-Mal at room temperature. The adsorption isotherm of Mal and Glu obtained at 22 degrees C shows the presence of an adsorption step at an average area/molecule of about 79 A2 between, approximately, 0.02 and 0.1 mM (the critical micelle concentration (cmc) is 2 mM) and 0.015 and 0.03 mM (the cmc is 2 mM), respectively. Similarly, for S Mal an adsorption plateau is observed at 70 A2 between 0.01 and 0.03 mM (the cmc is 0.7 mM). From the temperature dependence of the surface tension, we have seen that there are considerable differences in the adsorption of Glu and Mal. For Mal, the adsorption plateau is also observed at 29 degrees C at around 79 A2, whereas Glu exhibits no adsorption plateau at this temperature. At 8 degrees C, both Mal and Glu exhibit saturation behavior in the dilute part of the liquid expanded range, but at this temperature the average molecular areas are lower than at 22 degrees C: around 66 A2 for Glu and 75 A2 for Mal. Thus, the temperature sensitivity of Glu is considerably greater than for Mal in this range. The saturation regime coincides with a pronounced surface entropy minimum for Mal. The transition in the dilute liquid-expanded range supposedly occurs from a state with deformed surface micelles arranged in a hexagonal pattern, referred to as the granular range, to a true LE monolayer with a fluid hydrocarbon tail layer covering the entire surface. PMID- 15544383 TI - Adsorption of cobalt ions on the "electrolytic solution/gamma-alumina" interface studied by diffuse reflectance spectroscopy (DRS). AB - Diffuse reflectance spectroscopy was used for the first time to investigate the adsorption of the [Co(H2O)6]2+ ions on the interface developed between the surface of the gamma-alumina particles and the electrolytic aqueous solutions used for the preparation of cobalt-supported gamma-alumina catalysts by equilibrium deposition filtration. The formation of inner-sphere Co(II) surface complexes in which Co(II) is in octahedral symmetry was confirmed. A deconvolution peak centered at approximately 585 nm was attributed to the exchange of one aqua ligand with one AlxOHy (x = 1, 2, or 3; y = 0 or 1) negatively charged surface group resulting in the formation of mononuclear monosubstituted inner-sphere Co(II) complexes at a Co(II) surface concentration equal to 0.02 micromol of Co(II)/m2. It was inferred that as the surface Co(II) concentration increases the formation of disubstituted and/or trisubstituted surface complexes is favored with respect to the formation of monosubstituted Co(II) surface complexes. A deconvolution peak centered at approximately 640 nm was attributed to the exchange of one or more aqua ligands with bridging hydroxo ligands (Co-O-H). The relative magnitude of this peak increases with the Co(II) surface concentration, reflecting the increasing formation of binuclear, oligonuclear, and multinuclear Co(II) surface complexes and then the formation of the Co(II) surface precipitate. PMID- 15544385 TI - Permeable nonaggregating porphyrin thin films that display enhanced photophysical properties. AB - Porphyrins bearing bulky alkoxyphenyl substituents at two of the four meso positions and phenyl phosphonates at the other two have been prepared and used as building blocks for layer-by-layer assembly of conductive-glass-supported thin films via zirconium phosphonate chemistry. Thin-film characterization shows that the addition of sterically demanding 2,6-di(n-hexoxy)phenyl substituents to the meso-positions of the porphyrin skeleton can successfully prevent molecular aggregation. Both absorption and emission studies of multilayer thin films provide strong evidence that the new compounds have the ability to form thin films in which very little molecular (chromophore) interaction is present, relative to porphyrins that are not sterically hindered. Furthermore, the films are found to be permeable to selected small redox probes but blocking toward larger ones. Taken together, the sharp absorption spectra, increased emission yields, and permeability are expected to be advantageous for various materials based applications such as photovoltaics and sensors. PMID- 15544384 TI - Bonding and structure of glycine on ordered Al2O3 film surfaces. AB - The interaction between glycine (NH2CH2COOH) layers and an ultrathin Al2O3 film grown epitaxially onto NiAl(110) was studied by temperature-programmed desorption, X-ray photoelectron spectroscopy, ultraviolet photoelectron spectroscopy, work function measurements, and near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. At monolayer coverages at 110 K, there are two coexisting molecular forms: the anionic (NH2CH2COO-) and the zwitterionic form (NH3+CH2COO-) of glycine. As deduced from the photoemission data, the buildup of multilayers at 110 K leads to a condensed phase predominantly in the zwitterionic state. In contrast to the monolayer at 110 K, the monolayer formed at 300 K consists primarily of glycine molecules in the anionic state. The latter species is adsorbed with the oxygen atoms of the carboxylic group pointing toward the substrate. The polarization-dependent C K- and O K-edge NEXAFS spectra indicate that the glycinate species in the monolayer at 300 K is oriented nearly perpendicular to the surface, with the amino group pointing away from the surface. PMID- 15544386 TI - In situ formation and characterization of poly(ethylene glycol)-supported lipid bilayers on gold surfaces. AB - Inclusion of a polymer cushion between a lipid bilayer membrane and a solid surface has been suggested as a means to provide a soft, deformable layer that will allow for transmembrane protein insertion and mobility. In this study, mobile, tethered lipid bilayers were formed on a poly(ethylene glycol) (PEG) support via a two-step adsorption process. The PEG films were prepared by coadsorbing a heterofunctional, telechelic PEG lipopolymer (1,2-distearoyl-sn glycero-3-phosphoethanolamine-N-poly(ethylene glycol)-2000-N-[3-(2 (pyridyldithio)propionate]) (DSPE-PEG-PDP) and a nonlipid functionalized PEG-PDP from an ethanol/water mixture, as described in a previous paper (Munro, J. C.; Frank, C. W. Langmuir 2004, 20, 3339-3349). Then a two-step lipid adsorption strategy was used. First, lipids were adsorbed onto the PEG support from a hexane solution. Second, vesicles were adsorbed and fused on the surface to create a bilayer in an aqueous environment. Fluorescence recovery after photobleaching experiments show that this process results in mobile bilayers with diffusion coefficients on the order of 2 microm2/s. The mobility of the bilayers is decreased slightly by increasing the density of tethered lipids. The formation of bilayers, and not multilayer structures, is also confirmed by surface plasmon resonance, which was used to determine in situ film thickness, and by fluorimetry, which was used to determine quantitatively the fluorescence intensity for each 18 by 18 mm sample. Unfortunately, fluorescence microscopy also shows that there are large defects on the samples, which limits the utility of this system. PMID- 15544387 TI - Heterogeneity in styrene-butadiene latex films. AB - Low-Tg styrene-butadiene (SB) latex films were investigated by noncontact atomic force microscopy and scanning electric potential microscopy, revealing a number of different morphologies and electric potential patterns across films cast from the same SB latex dispersions under the same conditions. Surface leveling and charge dispersion throughout the films are, thus, restrained even at temperatures above Tg and the minimum film-formation temperature. An unprecedented electric pattern is observed, in which the particle cores are more positive than the contacting particle outer layers. Different packing patterns, including cubic and hexagonal arrays, coexist in neighboring areas. Zonal centrifugation of the SB latex in sucrose density gradient shows that particles cover a broad range of densities. Thus, film surface heterogeneity is at least partly due to particle heterogeneity. Fractal dimensions of topographic profiles are lower than those of the electric potential profiles, showing that charge mobility is much more restrained than polymer chain motion at the film surface and that it imposes a limit to the charged chain-ends motion. PMID- 15544388 TI - Control of photoreaction of amphiphilic spiropyran/n-alkane Langmuir and Langmuir Blodgett films using the phase transition of n-alkane. AB - The structures and photoreactions of Langmuir and Langmuir-Blodgett (LB) films of an amphiphilic spiropyran, 1',3'-dihydro-3',3'-dimethyl-6-nitro-1'-octadecyl-8 (docosanoyloxymethyl)spiro[2H-1-benzopyran-2,2'-(2H)-indole] (SP), mixed with n alkane are investigated. The mixing ratio was fixed at 1/2 for SP/n-alkane. The surface pressure-area isotherms of SP/octadecane are categorized into two regimes: a low-temperature regime where octadecane is packed with the alkyl chains of SP, and a high-temperature regime where the addition of octadecane does not influence the isotherms significantly. The temperature dividing the two regimes is related with the melting point of the n-alkane mixed with SP in the bulk. UV irradiation of the Langmuir film in the high-temperature regime gives rise to light-induced J-aggregation, whereas that in the low-temperature regime causes only the isomerization of SP to the corresponding merocyanine, indicating that J-aggregation is hindered by the presence of n-alkane in the low-temperature regime. IR external reflection spectroscopy of the Langmuir films shows that n alkane is released from the film during J-aggregation. The structural changes of the mixed Langmuir and LB films during J-aggregation are almost the same with those of the films of pure SP. PMID- 15544389 TI - Adsorption mechanism of substituted pyridines on silica suspensions: an NMR study. AB - Adsorption isotherms of 2,5-dimethylpyridine (2,5-DMP) on Aerosil 200 silica from water-2,5-DMP binary mixtures are known to exhibit special features indicative of surface phase transitions in the adsorbed layer. We have made similar observations on another substituted pyridine, 2,4,6-trimethylpyridine (2,4,6 TMP). By using 1D and 2D NMR spectroscopy, we investigated adsorbed layers on silica in suspension in water/substituted-pyridine mixtures and demonstrated the existence of adsorbed species specific signals. Comparison of signals with those displayed on NMR spectra of liquid binary mixtures under various pH conditions rules out adsorption via interaction of the surface silanol group and the lone electron pair of the nitrogen atom. A mechanism of adsorption through the interaction of surface siloxane oxygen and the aromatic pi-system is proposed; it is consistent with both thermodynamic measurements and stacking of substituted pyridines within the adsorbed layer. PMID- 15544390 TI - Probing macromolecular adsorbed layer structure and history dependence via the interfacial cavity function. AB - Adsorbed layers of proteins and other macromolecules often relax structurally more slowly than they form, rendering layer growth an out-of-equilibrium process. We show here how the interfacial cavity function, Phi (the average Boltzmann factor for a single probe molecule), may be determined, using kinetic data available from optical waveguide lightmode spectroscopy, and used as a continuous, in situ measure of history dependent adsorbed layer structure. The increase of Phi observed with residence time for fibronectin and lysozyme layers suggests post-adsorption clustering on a time scale longer than that predicted by a surface diffusion model. PMID- 15544391 TI - Effect of salt concentration on the nanostructure of weak polyacid brush in the amphiphilic polymer monolayer at the air/water interface. AB - The effect of salt concentration on the nanostructure of a spread monolayer of ionic amphiphilic diblock copolymer, (diethylsilacyclobutane)m-b-(methacrylic acid)n, at the air-water interface was directly investigated by in situ X-ray reflectivity and neutron reflectivity techniques. Previously, we had found that a poly(methacrylic acid) (PMAA) hydrophilic layer under the water was not in the form of a simple polyelectrolyte brush but consisted of a dense carpet upper layer and a diffuse brush lower layer when the hydrophilic chain was long enough. Here we observed this double layer formation in the monolayer in aqueous NaCl solution at a constant surface pressure. The effect of salt added to the subphase differed with the salt concentrations, that is, below or above 0.1 M. In the presence of NaCl up to 0.1 M, both the hydrophobic layer and brush layer thicknesses decreased. On the other hand, both of them increased in the presence of NaCl above 0.1 M. Also, the carpet layer thickness was almost constant independent of the salt concentration. In addition, the brush top roughness showed a maximum in the presence of 0.1 M NaCl. The increase of the charge number on the PMAA chain and the screening effect of the Coulomb interaction by added salt ions were considered to be responsible for these phenomena. PMID- 15544392 TI - FTIR studies of CO adsorption on Rh-Ge/Al2O3 catalysts prepared by surface redox reactions. AB - A series of bimetallic Al2O3-supported Rh-Ge catalysts was prepared by surface redox reactions under controlled hydrogen atmosphere. The surface properties of these catalysts were probed via in-situ FTIR spectroscopic studies of adsorbed CO and were compared to those of monometallic Rh catalysts that had undergone similar treatments. The results indicate that Ge addition results in the formation and stabilization of smaller rhodium ensembles at the expense of larger Rh0 surfaces. A charge-transfer mechanism from Ge to Rh is also inferred by the IR results for the high Ge loading samples. Air exposure of the catalysts leads to an irreversible segregation of the two metals and formation of large Rh crystallites. PMID- 15544393 TI - Effects of laterally heterogeneous slip on the resonance properties of quartz crystals immersed in liquids. AB - A formalism is presented which predicts the influence of laterally heterogeneous slip (for instance, induced by nanoscopic air bubbles) on the shift of the resonance frequency and bandwidth of quartz crystal resonators immersed in liquids. The lateral heterogeneities are decomposed into their Fourier components. The distribution of slip lengths provides a boundary condition, giving rise to a small, secondary flow field. The mean stress exerted by this secondary field induces a shift in resonance frequency and bandwidth. If the slip length is much smaller than the penetration depth of the shear waves and smaller than the lateral correlation length, one finds that the effects of the heterogeneities scale as n(3/2), with n being the overtone order. The frequency, f, and half-band-half-width, Gamma, decrease by the same amount. These calculations match experimental results obtained with a gold-coated resonator in contact with various hydrophilic liquids. PMID- 15544394 TI - Importance of molecular details in predicting bacterial adhesion to hydrophobic surfaces. AB - Electrostatic and hydrophobic forces are generally recognized as important in bacterial adhesion. Current continuum models for these forces often wrongly predict measurements of bacterial adhesion forces. The hypothesis tested here is that even qualitative guides to bacterial adhesion often require more than continuum information about hydrophobic forces; they require knowledge about molecular details of the bacteria and substrate surface. In this study, four different strains of bacteria were adsorbed to silica surfaces hydrophobized with alkylsilanes. The thickness of the lipopolysaccharide layers varied on the different bacteria, and the lengths of the alkylsilane molecules were varied from experiment to experiment. Bacterial adhesion was assessed using column experiments and atomic force microscopy (AFM) experiments. Results show that hydrophobized surfaces have higher bacterial sticking coefficients and stronger adhesion forces than bare silica surfaces, as expected. However, adhesion decreased as the solution Debye length became longer than the alkylsilane, perhaps since the silane molecules could not "reach" the bacterial surface. Similarly, those bacteria with a long o-antigen layer had decreased adhesion, perhaps since the silane molecules could not reach surface-bound proteins on the bacteria. This study reveals that macroscopic measurements such as contact angle are not able to fully describe bacterial adhesion; rather, additional details such as the molecular length are required to predict adhesion. PMID- 15544395 TI - Electrochemically programmed, spatially selective biofunctionalization of silicon wires. AB - A method for the spatially selective biofunctionalization of silicon micro- and nanostructures is reported, and results are presented for both single-crystal silicon (111) or (100) surfaces. An electroactive monolayer of hydroquinone was formed on the surface of H-terminated silicon working electrodes via an olefin reaction with UV-generated surface radicals. Molecules presenting either cyclopentadiene or a thiol group can be immobilized onto the regions where the hydroquinone has been oxidized. Molecular size and crystal orientation are evaluated as important factors that dictate the electrode stability in aqueous solution under anodic potentials. Monolayers composed of smaller molecules on (111) surfaces exhibit the highest packing density and are more effective in preventing anodic oxidation of the underlying substrate. Voltammetry, X-ray photoelectron spectroscopy, and atomic force and fluorescence microscopy are utilized to interrogate the kinetic rates of biofunctionalization, the extent of surface coverage, monolayer quality, and the spatial selectivity of the process. PMID- 15544396 TI - Protein adsorption onto silica nanoparticles: conformational changes depend on the particles' curvature and the protein stability. AB - We have analyzed the adsorption of protein to the surfaces of silica nanoparticles with diameters of 6, 9, and 15 nm. The effects upon adsorption on variants of human carbonic anhydrase with differing conformational stabilities have been monitored using methods that give complementary information, i.e., circular dichroism (CD), nuclear magnetic resonance (NMR), analytical ultracentrifugation (AUC), and gel permeation chromatography. Human carbonic anhydrase I (HCAI), which is the most stable of the protein variants, establishes a dynamic equilibrium between bound and unbound protein following mixture with silica particles. Gel permeation and AUC experiments indicate that the residence time of HCAI is on the order of approximately 10 min and slowly increases with time, which allows us to study the effects of the interaction with the solid surface on the protein structure in more detail than would be possible for a process with faster kinetics. The effects on the protein conformation from the interaction have been characterized using CD and NMR measurements. This study shows that differences in particle curvature strongly influence the amount of the protein's secondary structure that is perturbed. Particles with a longer diameter allow formation of larger particle-protein interaction surfaces and cause larger perturbations of the protein's secondary structure upon interaction. In contrast, the effects on the tertiary structure seem to be independent of the particles' curvature. PMID- 15544397 TI - pH-dependent mucoadhesion of a poly(N-isopropylacrylamide) copolymer reveals design rules for drug delivery. AB - This study investigated the mucoadhesive property of a hydrophobically modified copolymer N-isopropylacryamide and glycidylacrylamide NIPAM-N-Gly-(C18)2 (NIPAM Gly). Prior studies demonstrated that the interfacial properties of this copolymer are pH dependent and that the chains form strong hydrogen bonds at pH < 7 via the carboxylic acid side chains of the glycine moieties. Mucin interactions with the copolymer brushes were investigated by surface plasmon resonance and by direct force measurements. Mucin adsorption was determined as a function of pH, ionic strength, and mucin concentration. It adsorbs to the copolymer strongly at pH 5, but the adsorption decreases with increasing pH. The adsorbed amount is also ionic-strength dependent, decreasing with increasing monovalent salt concentrations at all pH values investigated. When compared with similar investigations with poly(ethylene oxide), these results provide insights into both the chemical characteristics and the solution conditions that determine the mucoadhesive properties of polymers. PMID- 15544398 TI - Self-assembly of trioxyethylene-encapsulated gold nanoclusters under aqueous conditions. AB - The aqueous self-assembly of methyl-terminated tri(oxyethylene)thiol-encapsulated gold nanoclusters of varying core size is demonstrated on micrometer scale Au/SiO2 interdigital electrodes. This self-assembly process consists of alternate exposures of the substrate to solutions of either an alpha,omega-dithiol or the gold nanoclusters, resulting in the deposition of these materials onto the electrode surface. A comparison of the procedure in both H2O and CHCl3 solvents shows that the assembly, as monitored by the electrical conductivity of the device, occurs more rapidly in the H2O system. This observation is complimented by XPS and UV/Vis measurements, which show that (1) the increased current is due to an increased amount of gold deposited on the surface under aqueous conditions and (2) the thiol exchange reaction occurs more rapidly in H(2)O in comparison to CHCl3. PMID- 15544399 TI - Monolayer binary active phase (Mo-V) and (Cr-V) supported on titania catalysts for the selective catalytic reduction (SCR) of NO by NH3. AB - Monolayer catalysts containing binary active phases (VOx-CrOx, VOx-MoOx) were prepared by simultaneous deposition of the corresponding transition metal-oxo species on the TiO2 (anatase) surface using the equilibrium deposition filtration technique. The prepared samples contained various amounts of each transition metal but almost the same total metal loading. They were characterized by atomic absorption spectroscopy, N2 adsorption, UV-vis diffuse reflectance spectroscopy, X-ray photoelectron spectroscopy, and tested for the selective catalytic reduction of NO by NH3 in the temperature range 250-450 degrees C. It was found that the transition-metal ionic species used for the preparation of these catalysts compete for the same surface sites of the TiO2 carrier upon co deposition. Small amounts of the second phase (Mo- or Cr-oxo phase) are sufficient in order to promote the catalytic activity at relatively high temperatures, in contrast to what happens in the corresponding industrial catalysts prepared by conventional methods. An electronic interaction between V- and Cr-oxo species favored at a V/Cr atomic ratio around 3 is probably responsible for the relatively high catalytic performance of the corresponding TiCrV catalyst. The activity of the studied catalysts is well correlated with the intensity of a DRS absorption band that appeared at ca. 400 nm, which is considered as a measure of the magnitude of interactions exerted between the monolayer transition metal-oxo species and the TiO2 carrier. This correlation is independent of the transition metals combination used and follows the same linear relationship found previously for single-active-phase catalysts. PMID- 15544400 TI - Preparation of poly(epsilon-caprolactone) brushes at the surface of conducting substrates. AB - This paper reports on the preparation of polyester brushes at the surface of electrically conducting materials. A two-step strategy has been worked out that consists of the electropolymerization of an acrylate under a cathodic potential, such that the polyacrylate layer is chemisorbed at the surface. In a second step, either preformed poly(epsilon-caprolactone) chains are grafted onto the polyacrylate sublayer or the ring-opening polymerization of epsilon-caprolactone is initiated from it. PMID- 15544401 TI - Humidity and temperature effects on CTAB-templated mesophase silicate films at the air-liquid interface. AB - Off-specular X-ray reflectivity measurements were carried out to follow the in situ development of surfactant-templated silica thin films at the air-water interface under conditions of controlled relative humidity and temperature, using an enclosed sample cell designed for this purpose. The results suggest a strong dependence of formation time and growth mechanism on ambient conditions. Thin films were synthesized at the air-water interface using cetyltrimethylammonium bromide (CTAB, 0.075 M) and a silica precursor, tetramethoxysilane (TMOS, 0.29 0.80 M) in an acidic medium. The studied humidity range was from 50 to 100%, the temperature was between 25 and 40 degrees C, and the TMOS/CTAB molar ratio was between 3.3 and 10.7. We observed that high humidity slows down the growth process due to lack of evaporation. However, increasing the temperature results in a decrease in the film-formation time. We proposed a formation mechanism for film growth as a consequence of phase separation, organic array assembly, and silica polymerization. PMID- 15544402 TI - Mechanical properties of polyelectrolyte-filled multilayer microcapsules studied by atomic force and confocal microscopy. AB - By using a combination of atomic force and confocal microscopy, we explore the deformation properties of multilayer microcapsules filled with a solution of strong polyelectrolyte. Encapsulation of polyelectrolyte was performed by regulation of the multilayer shell permeability in water-acetone solutions. The "filled"capsules prepared by this method were found to be stiffer than "hollow" ones, which reflects the contribution of the excess osmotic pressure to the capsule stiffness. The force-deformation curves contain three distinct regimes of reversible, partially reversible, and irreversible deformations depending on the degree of compression. The analysis of the shape of compressed capsules and of the inner polyelectrolyte spacial distribution allowed one to relate the deformation regimes to the permeability of the multilayer shells for water and inner polyelectrolyte at different stage of compression. PMID- 15544403 TI - Photoacoustic FT-IR depth imaging of polymeric surfaces: overcoming IR diffraction limits. AB - It is well established that the photoacoustic effect based on absorption of electromagnetic radiation into thermal waves allows surface depth profiling. However, limited knowledge exists concerning its spatial resolution. The spiral stepwise (SSW) approach combined with phase rotational analysis is utilized to determine surface depth profiling of homogeneous and nonhomogeneous multilayered polymeric surfaces in a step-scan photoacoustic FT-IR experiment. In this approach, the thermal wave propagating to the surface is represented as the integral of all heat wave vectors propagating across the sampling depth xn, and the spiral function K'beta(lambda)e(-beta)(lambda)xne(-x)n/mu(th)e(i)(omegat (xn/mu(th))) represents the amplitude and phase of the heat wave vector propagating to the surface. The SSW approach can be applied to heterogeneous surfaces by representing thermal waves propagating to the surface as the sum of the thermal waves propagating through homogeneous layers that are integrals of all heat vectors from a given sampling depth. The proposed model is tested on multilayered polymeric surfaces and shows that the SSW approach allows semiquantitative surface imaging with the spatial resolution ranging from micrometer to 500 nm levels, and the spatial resolution is a function of the penetration depth. PMID- 15544404 TI - Electroactive core-shell nanocluster films of heme proteins, polyelectrolytes, and silica nanoparticles. AB - Novel protein core-shell nanocluster films were assembled layer by layer on solid surfaces. In the first step, positively charged heme protein hemoglobin (Hb) or myoglobin (Mb) and negatively charged poly(styrenesulfonate) (PSS) were alternately adsorbed on the surface of SiO2 nanoparticles, forming core-shell SiO2-(protein/PSS)m nanoclusters. In the second step, the SiO2-(protein/PSS)m nanoclusters and polycationic poly(ethylenimine) (PEI) were assembled layer by layer on various solid substrates, forming [[SiO2-(protein/PSS)m]/PEI]n films. Various techniques were used to characterize the nanoclusters and monitor the film growth. [[SiO2-(protein/PSS)m]/PEI]n films at pyrolytic graphite (PG) electrodes exhibited well-defined, chemically reversible cyclic voltammetric reduction-oxidation peaks characteristic of the heme Fe(III)/Fe(II) redox couples. The proteins in the films retained near native conformations in the medium pH range, and the films catalyzed electrochemical reduction of oxygen and hydrogen peroxide. Advantages of the nanocluster films over the simple [SiO2/protein]n layer-by-layer films include a larger fraction of electroactive protein and higher specific biocatalytic activity. Using this approach, biocatalytic activity can be tailored and controlled by varying the number of bilayers deposited on the nanoparticle cores and the number of nanocluster layers on electrodes. PMID- 15544405 TI - Temperature-sensitive hybrid microgels with magnetic properties. AB - In the present paper, we report the preparation of hybrid temperature-sensitive microgels which include magnetite nanoparticles in their structure. Polymeric microgels have been prepared by surfactant-free emulsion copolymerization of acetoacetoxyethyl methacrylate (AAEM) and N-vinylcaprolactam (VCL) in water with a water-soluble azo-initiator. The obtained microgels possess a low critical solution temperature (LCST) in water solutions, with a rapid decrease of the particle size being observed at elevated temperatures. Magnetite was deposited directly into microgels, leading to the formation of composite particles which combine both temperature-sensitive and magnetic properties. The influence of magnetite load on microgel size, morphology, swelling-deswelling behavior, and stability is discussed. PMID- 15544406 TI - Competition of hydrogen-bonding and electrostatic interactions within hybrid polymer multilayers. AB - Using a layer-by-layer sequential adsorption technique, we report the construction of hybrid films in which layers of hydrogen-bonded polymers are embedded within electrostatically associated polyelectrolytes. The components of the hybrid film include a neutral hydrogen-bonding polymer, a weak polycarboxylic acid, and a strong polycation. Depending on the pH value used for the deposition of the electrostatic film, we found two distinctive regimes of film growth. At pHs lower than a critical value, deposition of electrostatic layers occurred on top of hydrogen-bonded stacks to produce hybrid, three-component films. At pHs higher than a critical value, neutral, hydrogen-bonded chains were displaced by the adsorbing chains of the polycation, producing two-component films. The property of the hydrogen-bonded stacks of hybrid films to be selectively dissolved by exposing them to a high pH makes these films promising candidates for producing free polyelectrolyte films. PMID- 15544407 TI - Synthesis of copolymer brushes endowed with adhesion to stainless steel surfaces and antibacterial properties by controlled nitroxide-mediated radical polymerization. AB - Novel copolymer brushes have been synthesized by a two-step "grafting from" method that consists of the electrografting of poly(2-phenyl-2-(2,2,6,6 tetramethyl-piperidin-1-yloxy)-ethylacrylate) onto stainless steel, followed by the nitroxide-mediated radical polymerization of 2-(dimethylamino ethyl)acrylate and styrene or n-butyl acrylate, initiated from the electrografted polyacrylate chains. The grafted copolymers were quaternized in order to endow them with antibacterial properties. Peeling tests have confirmed the strong adhesion of the grafted copolymer onto the stainless steel substrate. Quartz crystal microbalance experiments have proven that fibrinogen adhesion is lower on the hydrophilic quaternized films compared to the nonionic counterpart. Such quaternized copolymers exhibit significant antibacterial activity against the Gram-positive bacteria S. aureus and the Gram-negative bacteria E. coli. PMID- 15544408 TI - Layer-by-layer assembly of ordinary and composite coordination multilayers. AB - Coordination self-assembly of bishydroxamate-based metal-organic multilayers on gold employing a layer-by-layer (LbL) approach was investigated. It is shown that the solution chemistry of the participating metal ion has a marked influence on the composition and properties of the multilayers. Use of Ce4+ and particularly zirconium(IV) acetylacetonate (Zr(acac)4) solutions in the ion-binding step of multilayer construction leads to multilayers with a near-stoichiometric metal ion to-ligand ratio, suggesting a structure close to that predicted by a simple coordination self-assembly scheme. On the other hand use of a ZrCl4 solution as the source of metal ions in the multilayer construction leads to a multilayer with greater thickness and a large excess of Zr(IV), evenly distributed between the organic layers. In the latter case, a ratio of ca. 1:2 between the excess Zr and oxygen, as well as long-term Zr4+ binding experiments showing deposition of ZrO2, suggest the formation of a zirconia-type nanophase between the bishydroxamate organic repeat units during multilayer self-assembly. Hence, while the multilayer prepared using Zr(acac)4 solution appears to represent a "true" coordination-based structure, the one prepared using ZrCl4 is best described as a composite organic-ceramic multilayer. Composite multilayers prepared in this way display different properties from those of the stoichiometric ones, such as improved dielectric behavior and higher stiffness. Even greater mechanical stability is obtained with multilayers constructed using alternate binding of ZrCl4 and Ce4+. The concept of LbL formation of coordination-based composite organic-ceramic structures may be useful in obtaining nanometer-scale structures with tunable properties. PMID- 15544409 TI - Localized synthesis of polypyrrole in the nanopattern of monolayer films of diblock copolymer micelles. AB - A single-layered array of polystyrene-block-poly(4-vinylpyridine), PS-PVP, micelles in hexagonal order, fabricated by spin coating, was employed as a nanostructured template for synthesis of polypyrrole, a conducting polymer, in nanometer-sized domains. Oxidative catalysts of FeCl3 for the polymerization were selectively loaded in spherical PVP nanodamains so that they were hexagonally arranged over the film but confined in the nanometer range. The vapor-phase polymerization of pyrrole was localized in the PVP nanodomains, leading to a morphological transition from spherical to wormlike domains. In addition, the nanodomains containing polypyrrole were converted to open cavities by ethanol, a PVP block-selective solvent. PMID- 15544410 TI - Mesomorphous structure and properties of non-equimolar complexes of poly(ethylenimine) and perfluorooctanoic acid. AB - A series of solid complexes, PEI-PFAO, made of poly(ethylenimine) (PEI) and perfluorooctanoic acid (PFOA) with different compositions were prepared through a "starving addition" method, where PFOA was fed into PEI solution at the molar ratio, phi(feed), of acid group to the amino group of PEI, never beyond unity. Wide-angle X-ray diffraction diagrams confirmed amorphous structure of these complexes. Small-angle X-ray scattering indicated two ordered mesomorphous structures of alpha and beta lamellar phases, with respective long periods of 2.29 and 1.15 nm in the complexes. By increasing the actual molar ratio, phi, of PFOA to the amino group of PEI, the complex structure was altered from alpha phase dominant to beta-phase dominant. All complexes exhibited two thermal degradation processes induced by decomposition of the bound PFOA below 230 degrees C and PEI backbone at about 350 degrees C. The initiating degradation temperature, Tid, decreases with increasing phi due to the preferential degradation of the PFOA chain bound to the tertiary amino groups. The glass transition temperature, Tg, of the complex increases with phi up to the degradation of the complex of phi = 1. This increase in Tg with phi also supports an ordered alignment of the bound PFOA chains, which greatly restricts the PEI mobility. The solid surface tension, gammaS, and critical surface tension, gammac, of the complex are between 15.4 and 16.8 mN/m and between 13.5 and 15.4 mN/m, respectively. The latter is very close to or even smaller than gammac of PTFE (15 mN/m), suggesting the enrichment of CF2 and CF3 groups at the complex surfaces. The fact that the PEI-POFA complex combines high hydrophobicity with selective thermal degradation of bound fluorinated chains promises a potential of selective change and local functionalization of the surface in a well-controlled manner. PMID- 15544411 TI - Dielectrophoretic forces on the nanoscale. AB - We have developed a method of calculation of the dielectrophoretic force on a nanoparticle in a fluid environment where variations in the electric field and electric field gradients are on the same nanoscale as the particle. The Boundary Element Dielectrophoretic Force (BEDF) method involves constructing a solvent accessible or molecular surface surrounding the particle, calculating the normal component of the electric field at the surface boundary elements, and then solving a system of linear equations for the induced surface polarization charge on each element. Different surface elements of the molecule may experience quite different polarizing electric fields, unlike the situation in the point dipole approximation. A single 100-A-radius ring test configuration is employed to facilitate comparison with the well-known point dipole approximation (PDA). We find remarkable agreement between the forces calculated by the BEDF and PDA methods for a 1 A polarizable sphere. However, for larger particles, the differences between the methods become qualitative as well as quantitative; the character of the force changes from attractive at the origin of the ring for a 50 A sphere, to repulsive for a 75-A sphere. Equally dramatic differences are found in a more complex electrical environment involving two sets of 10 rings. PMID- 15544412 TI - Comparative study of Li, Na, and K adsorptions on graphite by using ab initio method. AB - A comprehensive study has been conducted to compare the adsorptions of alkali metals (including Li, Na, and K) on the basal plane of graphite by using molecular orbital theory calculations. All three metal atoms prefer to be adsorbed on the "middle hollow site" above a hexagonal aromatic ring. A novel phenomenon was observed, that is, Na, instead of Li or K, is the weakest among the three types of metal atoms in adsorption. The reason is that the SOMO (single occupied molecular orbital) of the Na atom is exactly at the middle point between the HOMO and the LUMO of the graphite layer in energy level. As a result, the SOMO of Na cannot form a stable interaction with either the HOMO or the LUMO of the graphite. On the other hand, the SOMO of Li and K can form a relatively stable interaction with either the HOMO or the LUMO of graphite. Why Li has a relatively stronger adsorption than K on graphite has also been interpreted on the basis of their molecular-orbital energy levels. PMID- 15544413 TI - Ground-state clusters for short-range attractive and long-range repulsive potentials. AB - We report calculations of the ground-state energies and geometries for clusters of different sizes (up to 80 particles), where individual particles interact simultaneously via a short-ranged attractive potential, modeled with a generalization of the Lennard-Jones potential, and a long-ranged repulsive Yukawa potential. We show that for specific choices of the parameters of the repulsive potential, the ground-state energy per particle has a minimum at a finite cluster size. For these values of the parameters in the thermodynamic limit, at low temperatures and small packing fractions, where clustering is favored and cluster cluster interactions can be neglected, thermodynamically stable cluster phases can be formed. The analysis of the ground-state geometries shows that the spherical shape is marginally stable. In the majority of the studied cases, we find that above a certain size, ground-state clusters preferentially grow almost in one dimension. PMID- 15544414 TI - Monte Carlo molecular simulation of the hydration of K-montmorillonite at 353 K and 625 bar. AB - Monte Carlo molecular simulations of the hydration of K-saturated Wyoming-type montmorillonite at constant stress in the NPzzT ensemble and at constant chemical potential in the grand canonical muVT ensemble, under basin-like conditions of 353 K and 625 bar, show a strong tendency of the K+ ions to adhere to the siloxane surface, forming predominant inner-sphere complexes with tetrahedral oxygen atoms and adsorbed water molecules. Simulations in the grand canonical ensemble predict that none of the K-montmorillonite hydrates, the one-, two-, and three-layer hydrates, are stable in this environment of high depth, temperature, and pressure. The most nearly stable configuration corresponds to the one-layer hydrate, characterized by a d001 spacing of 12.75 A, the adsorbed water being 60 molecules/layer or 180.83 mg of H2O/g of clay, an internal energy of -22.73 kcal/mol, an interlayer density of 0.365 g/mL, and a pressure tensor, Pzz, of 1999.9 bar. The interlayer structure consists of two close layers of water molecules 0.50 A from the midplane, with broad shoulders on the sides, the protons oriented toward the midplane and the siloxane surfaces, and the K+ ions close to the clay surfaces and on the interlayer midplane. PMID- 15544415 TI - Superhydrophobic, high surface area, 3-D SiO2 nanostructures through siloxane based surface functionalization. PMID- 15544416 TI - Light and the behavior of ciliates. PMID- 15544417 TI - Effect of ischemia and reperfusion on protein oxidation in isolated rabbit hearts. AB - Reactive oxygen species and other oxidants are involved in the mechanism of postischemic contractile dysfunction, known as myocardial stunning. The present study investigated the oxidative modification of cardiac proteins in isolated Langendorff-perfused rabbit hearts subjected to 15 min normothermic ischemia followed by 10 min reperfusion. Reperfusion under these conditions resulted in only 61.8+/-2.7 % recovery of developed pressure relative to preischemic values and this mechanical dysfunction was accompanied by oxidative damage to cardiac proteins. The total sulfhydryl group content was significantly reduced in both ventricle homogenates and mitochondria isolated from stunned hearts. Fluorescence measurements revealed enhanced formation of bityrosines and conjugates of lipid peroxidation-end products with proteins in cardiac homogenates, whereas these parameters were unchanged in the mitochondrial fraction. Reperfusion did not alter protein surface hydrophobicity, as detected by a fluorescent probe 1 anilino-8-naphthalenesulfonate. Our results indicate that oxidation of proteins in mitochondria and possibly in other intracellular structures occurs during cardiac reperfusion and might contribute to ischemia-reperfusion injury. PMID- 15544418 TI - Decrease in serum dehydroepiandrosterone level after fenofibrate treatment in males with hyperlipidemia. AB - The influence of steroid hormones on plasma lipids and lipoproteins was confirmed by many studies. On the other hand, the effect of plasma lipids on metabolism of steroid hormones has so far not been examined. The objective of this research project was to determine (1) the levels of cortisol, testosterone, estradiol, dehydroepiandrosterone (DHEA), its sulfate (DHEAS), 7-hydroxylated DHEA, and SHBG in men suffering from mixed hyperlipidemia (HPL) (n=23, age 46.1+/-7.9 years) in comparison with healthy male volunteers (n=17, age 45.1+/-15.6 years); (2) whether therapy with fenofibrate influences the levels of the above mentioned steroids and SHBG; (3) what are the correlations between lipids and steroids in healthy males and HPL patients before and after therapy. Compared to controls, untreated patients had significantly higher estradiol and free testosterone index (IFT) levels (p<0.0003 and p<0.02, respectively) and significantly lower SHBG (p<0.02). Due to fenofibrate therapy, a significant decrease of TC, TG, and DHEA levels occurred (mean decrease: 14 %, 52 % and 21 %, respectively). Triglycerides correlated negatively with testosterone and SHBG in healthy subjects. HDL-C correlated positively and consequently, atherogenic index correlated negatively with 7-hydroxylated epimers of DHEA in treated patients. This is the first study dealing with the influence of fenofibrate administration on the steroid levels. Taking together, the most important is the finding of decrease DHEA levels after fenofibrate therapy. It could be explained, at least in part, by the effect of the fenofibrateon on the biosynthesis of DHEA and its regulation. PMID- 15544419 TI - Influence of physiological changes of glycaemia on VEPs and visual ERPs. AB - Since hypoglycemia is known to influence cognitive functions, we checked whether the physiological changes in glycemia (after fasting or exertion) can explain the rather high intra-individual variability of event-related potentials (ERPs). Besides the ERPs to "change in coherence of a moving pattern" with reaction time (RT) recording, binocular pattern reversal VEPs and motion-onset VEPs (to linear and radial motion) were also examined in 14 healthy subjects prior to and after 24-h fasting that decreased glycemia from 5.3 to 3.9 mmol/l on the average. We only found one significant change in the latencies and amplitudes of VEPs and ERPs (with no change of RT). The N160 peak in the motion-onset VEPs to radial (expansive) motion (EM-VEPs) showed a larger amplitude at lower glycemia. For evaluation of the exertion influence, we tested glycemia prior to and after 90 min long exercise -- bicycle ergometry with the load set to 2 W/kg in women and 2.5 W/kg in men (average age-related values for W170/kg index). The changes of glycemia to exertion were, however, less distinct than those to fasting. We conclude that in healthy subjects the glycemia decrease due to 24-h fasting or intensive time-limited exercise never reaches the critical value to change the VEP, ERPs and RTs. PMID- 15544420 TI - Influence of magnesium sulphate on evoked activity of rat brain after exposure to short-term hypoxia. AB - Young Wistar rats (aged 12, 25 and 35 days) were exposed to short-term (60 min) hypobaric hypoxia of 41 kPa. Cortical afterdischarges (ADs) were evoked by repeated direct stimulation of the sensorimotor cortex and the duration of ADs was monitored to examine the influence of magnesium sulphate injection (0.3 g/kg b.w.). In 12-day-old hypoxia-exposed rats, an increase of the mean duration of ADs after the repeated stimulation appeared. This effect was prevented by magnesium administration. In 25- and 35-day-old rats exposed to hypoxia a shortening of ADs was registered but no specific effect of magnesium sulphate pretreatment was observed. The brain susceptibility and ability to terminate evoked seizures is discussed. PMID- 15544421 TI - The effect of tryptophan depletion on brain activation measured by functional magnetic resonance imaging during the Stroop test in healthy subjects. AB - We investigated the role of serotonin in cognitive activation of the frontal cortex. The serotonergic system was affected by the administration of an amino acids mixture without tryptophan (tryptophan depletion). In a placebo-controlled double-blind cross-over study with 20 healthy volunteers, we tested the hypothesis that a tryptophan (serotonin) decrease affects the activation of prefrontal cortex by the Stroop test. Cognitive brain activation was evaluated by functional magnetic resonance imaging (fMRI). Tryptophan depletion decreased the plasma tryptophan level up to 90 % for five hours after the tryptophan-free drink had been consumed when compared with the same mixture with tryptophan (p?0.0001). Tryptophan depletion did not affect the Stroop test performance. We compared fMRI activation in both conditions (tryptophan depletion and placebo) with plasma tryptophan levels as the covariates. The tryptophan depletion increased the activation (fMRI signal) in the bilateral mediofrontal cortex, anterior cingulate and left dorsolateral prefrontal cortex. The present findings allow the postulate that serotonergic medial forebrain and cingulum bundle pathways play a role in the activity of cortical structures involved in Stroop test processing. PMID- 15544422 TI - Action of granulopoiesis-stimulating cytokines rhG-CSF, rhGM-CSF, and rmGM-CSF on murine haematopoietic progenitor cells for granulocytes and macrophages (GM-CFC). AB - The aim of this study was to provide new data to the knowledge of mechanisms by which recombinant human granulocyte colony-stimulating factor (rhG-CSF), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) enhance the numbers of colonies growing from hematopoietic progenitor cells for granulocytes and macrophages (GM-CFC) in the murine bone marrow. The in vitro technique for cultivating GM-CFC from normal bone marrow cells was used. For evaluation of stimulatory actions of the drugs studied, the factors themselves or sera of mice given these factors were added to the cultures. The factors or the sera were present in the cultures either as the only potentially stimulatory agents or acted jointly with a suboptimum concentration of recombinant murine interleukin-3 (rmIL-3). It was found that both rhG-CSF and rmGM-CSF stimulate the proliferation of GM-CFC by a combination of direct mechanisms (direct actions on the target cells) and indirect effects (effects mediated through the induction of other cytokines and/or growth factors in the murine organism). The rhGM-CSF exhibited somewhat weaker in vitro effects in comparison with the other two factors and only indirect effects were noted. Additional in vivo experiments documented that, in spite of differences in mechanisms of action of the individual drugs studied on murine bone marrow cells in vitro, equal in vivo doses of the factors induce quantitatively similar effects on the production of GM-CFC in vivo. PMID- 15544423 TI - Relations between particle size of HDL and LDL lipoproteins and cholesterol esterification rate. AB - Particle size of low density (LDL) and high density (HDL) lipoproteins and cholesterol esterification rate in HDL plasma (FER(HDL)) are important independent predictors of coronary artery diseases (CAD). In this study we assessed the interrelations between these indicators and routinely examined plasma lipid parameters and plasma glucose concentrations. In 141 men, healthy volunteers, we examined plasma total cholesterol (TC), triglycerides (TG), HDL and LDL cholesterol (HDL-C, LDL-C) and HDL unesterified cholesterol (HDL-UC). Particle size distribution in HDL and LDL was assessed by gradient gel electrophoresis and FER(HDL) was estimated by radioassay. An effect of particle size and FER(HDL) on atherogenic indexes as the Log(TG/HDL-C) and TC/HDL-C was evaluated. Subjects in the study had plasma concentrations (mean +/- S.D.) of TC 5.2+/-0.9 mmol/l, HDL-C 1.2+/-0.3 mmol/l, TG 2.1+/-1.7 mmol/l, glucose 5+/-0.8 mmol/l. Relative concentration of HDL(2b) was 17.6+/-11.5 % and 14.6+/-11.8 % of HDL(3b,c). The mean diameter of LDL particles was 25.8+/-1.5 nm. The increase in FER(HDL) significantly correlated with the decrease in HDL(2b) and LDL particle size (r = -0.537 and -0.583, respectively, P<0.01) and the increase in HDL(3b,c) (0.473, P<0.01). Strong interrelations among TG and HDL-C or HDL-UC and FER(HDL) and particle size were found, but TC or LDL-C did not have such an effect. Atherogenic indexes Log(TG/HDL-C) and TC/HDL-C correlated with FER(HDL) (0.827 and 0.750, respectively, P<0.0001) and with HDL and LDL particle size. PMID- 15544424 TI - The computer modelling of human TRH receptor, TRH and TRH-like peptides. AB - The aim of this work was to verify the possibility of interactions between the human TRH receptor (an integral membrane protein which belongs to family 1 of G protein coupled receptors) and TRH-like peptides presented in the prostate gland. These peptides are characterized by substitution of basic amino acid histidine (related to authentic TRH) for neutral or acidic amino acid, such as glutamic acid, phenylalanine, glutamine or tyrosine. The physiological function of TRH like peptides in peripheral tissues is not precisely known. However, according to our recent experiments, we assume the existence of a local hormonal network formed by TRH-like peptides and TSH in the prostate gland. The network can be associated with circulating thyroid and steroid hormones, and may represent a new regulatory mechanism influencing the proliferative ability of prostatic tissue. A similar network of authentic TRH and TSH was already found in the gastrointestinal tract. The experimentally determined 3D-structures of human TRH receptor (hTRHr) and TRH-like peptides are not available. From this point of view we used de novo modeling procedures of G-protein coupled receptors on an automated protein modeling server used at the Glaxo Wellcome Experimental Research (Geneva, Switzerland). 3D-structures of TRH-like peptides were determined with a computer program CORINA (written by the team of J. Gasteiger, Computer-Chemie-Centrum and Institute for Organic Chemistry, University of Erlangen-Nurenberg, Germany). The generated PDB files with 3D-coordinates were visualized with Swiss-Pdb Viewer Release 3.51 (Glaxo Wellcome). From recent results it is evident that polar amino acids belonging to the extracellular terminus of hTRHr transmembrane regions can participate in interactions between TRH and hTRHr. There is no direct evidence that TRH-like peptides interact with the presented hTRHr model. On the contrary, with respect to the similar 3D-shape and the identity of terminal amino acids, it appears that these interactions are highly probable as well as the nearly 100 % cross-reactions between TRH or TRH like peptides and antibody specific against authentic TRH. Closed terminal amino acids (pyroglutamic acid and proline-amide) of TRH or TRH-like peptides are important for these interactions. Desamido-TRH or glutamyl metabolites will be repelled by the negative potential of ASP195 (E: D93) and GLU298 (G: E137). PMID- 15544425 TI - Effects of a free radical scavenger N-tert-butyl-alpha-phenylnitrone (PBN) on short-term recovery of immature rats after status epilepticus. AB - The present study examined the effects of a free radical scavenger, N-tert-butyl alfa-phenylnitrone (PBN) on lithium-pilocarpine-induced status epilepticus (SE) and its short-term consequences in rats 12 (P12) or 25 (P25) days old. PBN (2 x 100 mg/kg i.p.) was injected according to the following schedules: 1) PBN pretreated animals received the first dose 30 min prior to pilocarpine, the second dose was given 1 min after SE onset, and 2) PBN-treated animals received the first dose of PBN 1 min after SE onset and the second one 60 min later. Paraldehyde was administered to decrease mortality. Effects of PBN were highly age-dependent. In P25 group, PBN-pretreatment increased latency to SE onset and significantly suppressed the severity of motor manifestation of SE. Both PBN pretreatment and treatment improved recovery after SE. In contrast, administration of PBN in P12 animals did not affect SE pattern or recovery after SE. Administration of PBN had no effects on the motor performance of animals 3 and 6 days after SE. Neuronal damage was examined 24 h and 7 days after SE using Fluoro-Jade B staining. Mild neuroprotective effects of PBN in hippocampal fields CA1 and CA3 occurred in P25 rats in both experimental schedules. In contrast, administration of PBN aggravated neuronal injury in the hippocampus in P12 rats. Administration of PBN to intact rats did not induce neurodegeneration in either age group. PMID- 15544426 TI - Adiponectin, an adipocyte-derived protein. AB - Adipose tissue is a hormonally active tissue, producing adipocytokines which may influence activity of other tissues. Adiponectin, abundantly present in the plasma increases insulin sensitivity by stimulating fatty acid oxidation, decreases plasma triglycerides and improves glucose metabolism. Adiponectin levels are inversely related to the degree of adiposity. Anorexia nervosa and type 1 diabetes are associated with increased plasma adiponectin levels and higher insulin sensitivity. Decreased plasma adiponectin levels were reported in insulin-resistant states, such as obesity and type 2 diabetes and in patients with coronary artery disease. Activity of adiponectin is associated with leptin, resistin and with steroid and thyroid hormones, glucocorticoids, NO and others. Adiponectin suppresses expression of extracellular matrix adhesive proteins in endothelial cells and atherosclerosis potentiating cytokines. Anti-atherogenic and anti-inflammatory properties of adiponectin and the ability to stimulate insulin sensitivity have made adiponectin an important object for physiological and pathophysiological studies with the aim of potential therapeutic applications. PMID- 15544427 TI - Leptin, soluble leptin receptor and leptin gene polymorphism in relation to preeclampsia risk. AB - Few investigators have simultaneously evaluated leptin, soluble leptin receptor (SLR) and leptin gene polymorphisms in preeclampsia cases and controls. We examined these three biomolecular markers in 40 preeclampsia cases and 39 controls. Plasma leptin and SLR concentrations were determined using immunoassays. Genotype for the tetranucleotide repeat (TTTC)(n), polymorphism in the 3 -flanking region of the leptin gene was determined using PCR. Alleles of the polymorphism were characterized by size distributions [short repeats (class I); and long repeats (class II)]. Logistic regression was used to calculate odds ratios (OR) and 95 % confidence intervals (CI). Leptin concentrations were higher in our cases than in the controls (53.1 4.7 vs. 17.7+/-2.4 ng/ml, p<0.05). SLR concentrations were slightly lower in our patients than in the controls (25.7+/ 1.9 vs. 29.1+/-1.1 ng/ml, p>0.05). Elevated leptin (? 14.5 ng/ml) was associated with a 3.8-fold (CI 1.0-14.4) increased risk; whereas low SLR (< 28.5 ng/ml) was associated with a 6.3-fold (CI 1.7-23.2) increased risk of preeclampsia. The I/II genotype was associated with a 3.8-fold increased risk of preeclampsia (OR=3.8; 95 % CI 0.8-18.0); and the II/II genotype was not observed among our cases (0 % vs. 33 % p<0.001). Larger studies would be needed to confirm and further clarify the relations between functional variants in the leptin gene and preeclampsia risk. PMID- 15544428 TI - Effects of Cu/Zn superoxide dismutase on strain injury-induced oxidative damage to skeletal muscle in rats. AB - This study was designed to determine whether the supplement of superoxide dismutase (SOD) could attenuate strain-induced oxidative damage to skeletal muscle in rats. Experimental animals were injured in right gastrocnemius muscles by a strain injury model. SOD-treated groups were given Cu/Zn SOD 10 000 U/kg body weight per day since injured, while control groups were given normal saline. Parameters of antioxidant and muscle damage were detected in plasma 3 and 7 days postinjury. The injured muscles were removed and fixed for histology observation and immunohisto-chemistry assay of desmin. The results showed that plasma levels of SOD, glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC) in SOD group were significantly higher than in the saline group on day 3 or 7, while the plasma creatine kinase (CK) and malondialdehyde (MDA) were lower in the SOD group than in the saline group. The histological examination of muscle sections revealed a lower degree of damage in the SOD group in which the expression level of desmin was higher than in the saline group. It is suggested that SOD supplement may attenuate strain-induced muscle damage and facilitate its regeneration. PMID- 15544429 TI - Effect of normoxia and hypoxia on K(+) current and resting membrane potential of fetal rabbit pulmonary artery smooth muscle. AB - At birth, the increase in O(2) tension (pO(2)) is an important cause of the decrease in pulmonary vascular resistance. In adult animals there are impressive interspecies differences in the level of hypoxia required to elicit a pulmonary vasoconstrictor response and in the amplitude of the response. Hypoxic inhibition of some potassium (K(+)) channels in the membrane of pulmonary arterial smooth muscle cells (PASMCs) helps to initiate hypoxic pulmonary vasoconstriction. To determine the effect of the change in pO(2) on fetal rabbit PASMCs and to investigate possible species-dependent differences, we measured the current voltage relationship and the resting membrane potential, in PASMCs from fetal resistance arteries using the amphotericin-perforated patch-clamp technique under hypoxic and normoxic conditions. Under hypoxic conditions, the K(+) current in PASMCs was small, and could be inhibited by 4-aminopyridine, iberiotoxin and glibenclamide, reflecting contributions by Kv, K(Ca) and K(ATP) channels. The average resting membrane potential was -44.3+/-1.3 mV (n=29) and could be depolarized by 4-AP (5 mM) and ITX (100 nM) but not by glibenclamide (10 microM). Changing from hypoxia, that mimicked fetal life, to normoxia dramatically increased the K(Ca) and consequently hyperpolarized (-9.3+/-1.7 mV; n=8) fetal rabbit PASMCs. Under normoxic conditions K(+) current was reduced by 4-AP with a significant change in resting membrane potential (11.1+/-1.7 mV; n=8). We conclude that resting membrane potential in fetal rabbit PASMCs under both hypoxic and normoxic conditions depends on both Kv and K(Ca) channels, in contrast to fetal lamb or porcine PASMCs. Potential species differences in the K(+) channels that control resting membrane potential must be taken into consideration in the interpretation of studies of neonatal pulmonary vascular reactivity to changes in O(2) tension. PMID- 15544431 TI - Cigarette smoking, metabolic activation and carcinogenesis. AB - Epidemiologically, it has been suggested that cigarette smoking is closely associated with an increased risk of cancers in various organs such as the lung, oropharynx, stomach, pancreas, liver and colon. Nevertheless, influences of cigarette smoking on experimental tumorigenesis in organs other than the respiratory tract remain to be elucidated. In our experimental studies, it has been shown that cigarette smoke exposure induces hepatic CYP enzymes, especially CYP1A2, in both rats and hamsters, and S9 fraction from their livers exposed to cigarette smoke specifically increases the mutagenicity in Ames assay of various heterocyclic amines (HCAs) contained in cigarette smoke and cooked food, which is in good agreement with the fact that HCAs are principally activated by CYP1A2 to proximate carcinogens. In fact, cigarette smoke exposure enhanced liver carcinogenesis in rats induced by 2-amino-3, 8-dimethylimidazo[4, 5-f]quinoxaline (MeIQx), a major HCA. Furthermore, in our recent study, it was also shown that cigarette smoke exposure induces hepatic CYP2A8 in hamsters, which is homologous to CYP2A6 in human, and hepatic S9 fraction exposed to cigarette smoke increases the mutagenicity of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a tobacco specific nitrosamine, which is in line with the fact that NNK is metabolically activated by CYP2A6. Keeping these data, the aim of this review is to discuss any relevancy of modulated metabolic activation by cigarette smoking to cancer risk in human. PMID- 15544432 TI - Biopharmaceutic classification system: a scientific framework for pharmacokinetic optimization in drug research. AB - The tenets of biopharmaceutics, solubility and permeability, are of pivotal importance in new drug discovery and lead optimization due to the dependence of drug absorption and pharmacokinetics on these two properties. A classification system for drugs based on these two fundamental parameters, Biopharmaceutic Classification System (BCS), provides drug designer an opportunity to manipulate structure or physicochemical properties of lead candidates so as to achieve better "deliverability". Considering the facts for failure of NCEs, drug research, once concentrating on optimizing the efficacy and safety of the leads, dramatically transformed in the past two decades. With the enormous number of molecules being synthesized using combinatorial and parallel synthesis, high throughput methodologies for screening solubility and permeability has gained significant interest in pharmaceutical industry. Ultimate aim of the drug discovery scientist in pharmacokinetic optimization is to tailor the molecules so that they show the features of BCS class I without compromising on pharmacodynamics. Considerations to optimize drug delivery and pharmacokinetics right from the initial stages of drug design propelled need for "High Throughput Pharmaceutics" (HTP). In silico predictions and development of theoretical profiles for solubility and lipophilicity provides structure based biopharmaceutical optimization, while in vitro experimental models (microtitre plate assays and cell cultures) validate the predictions. Thus, biopharmaceutical characterization during drug design and early development helps in early withdrawal of molecules with insurmountable developmental problems associated with pharmacokinetic optimization. PMID- 15544433 TI - Metabonomics: its potential as a tool in toxicology for safety assessment and data integration. AB - The functional genomic techniques of transcriptomics and proteomics promise unparalleled global information during the drug development process. However, if these technologies are used in isolation the large multivariate data sets produced are often difficult to interpret, and have the potential of missing key metabolic events (e.g. as a result of experimental noise in the system). To better understand the significance of these megavariate data the temporal changes in phenotype must be described. High resolution 1H NMR spectroscopy used in conjunction with pattern recognition provides one such tool for defining the dynamic phenotype of a cell, organ or organism in terms of a metabolic phenotype. In this review the benefits of this metabonomics/metabolomics approach to problems in toxicology will be discussed. One of the major benefits of this approach is its high throughput nature and cost effectiveness on a per sample basis. Using such a method the consortium for metabonomic toxicology (COMET) are currently investigating approximately 150 model liver and kidney toxins. This investigation will allow the generation of expert systems where liver and kidney toxicity can be predicted for model drug compounds, providing a new research tool in the field of drug metabolism. The review will also include how metabonomics may be used to investigate co-responses with transcripts and proteins involved in metabolism and stress responses, such as during drug induced fatty liver disease. By using data integration to combine metabolite analysis and gene expression profiling key perturbed metabolic pathways can be identified and used as a tool to investigate drug function. PMID- 15544434 TI - Effect of growth factors as therapeutic drugs on hepatic metabolism during the systemic inflammatory response syndrome. AB - Characteristic for critically ill patients is a hypermetabolism and catabolism that is associated with impairment of the structure and function of essential organs, such as the immune system, kidney, peripheral muscles and the liver. The liver-gut-axis, with liver integrity, metabolism and function are crucial for survival of patients suffering from trauma, operations or infections. The hepatic acute phase response represents a cascade of events characterized by the upregulation of acute phase proteins and the downregulation of constitutive hepatic proteins. The goal of the hepatic acute-phase-response, which is mediated by cytokines and signal transcription factors, is to restore homeostasis. However, multiple studies have shown that a sustained or increased acute phase response is detrimental with the uncontrolled and prolonged action of acute phase proteins. The downregulation of constitutive hepatic proteins may further augment these detrimental effects. Research has focused on the attenuation of the inflammatory response using anti-inflammatory agents or antibodies against pro inflammatory cytokines such as tumor necrosis (TNF), interleukin-1beta (IL 1beta), or their receptors. These approaches showed promising results in vitro and in animal models; however, when these approaches entered clinical trials it became evident that these promising animal data could not be substantiated in humans. A different approach is to attenuate the inflammatory cascade by the administration of growth factors. Growth factors exert anabolic effects and affect the inflammatory hepatic metabolism. The present review discusses the effect of recombinant human growth hormone, insulin-like growth factor-I, hepatocyte growth factor and insulin on the hepatic metabolism and homeostasis during inflammation and delineates the therapeutic benefit and limitation of growth factor administration in critically ill patients. PMID- 15544435 TI - Therapeutic drugs that behave as mechanism-based inhibitors of cytochrome P450 3A4. AB - Cytochrome P450 (CYP) 3A4 is not only the most abundant isoform in human liver but also metabolizes approximately 60% of the therapeutic drugs. This feature renders CYP3A4 highly susceptible to both reversible and irreversible (mechanism based) inhibition. The latter is characterized by NADPH-, time- and concentration dependent enzyme inactivation, occurring when some drugs are converted by CYPs to reactive metabolites. Mechanism-based inactivation of CYP3A4 by drugs can be due to the chemical modification of the heme, the protein, or both as a result of covalent binding of modified heme to the protein. The clinical pharmacokinetic effect of a CYP3A4 inactivator is a function of its KI, kinact and partition ratio and the synthesis rate of new or replacement enzyme. Predicting drug-drug interactions involving CYP3A4 inactivation is possible when proper pharmacokinetic principles are followed. However, the prediction may become difficult, since the clinical outcomes due to CYP3A4 inactivation depend on many factors associated with the enzyme, drugs and the patients. A number of clinically important drugs have been identified to be mechanism-based CYP3A4 inhibitors. These include antibiotics (e.g. erythromycin and isoniazid), anticancer drugs (e.g. tamoxifen), antidepressants (e.g. fluoxetine and midazolam), anti-HIV agents (e.g. ritonavir and delavirdine), antihypertensives (e.g. dihydralazine and verapamil), steroids and their receptor modulators (e.g. gestodene and raloxifene), and some herbal constituents (e.g. bergamottin and glabridin). Compared to reversible inhibition, mechanism-based inhibitors of CYP3A4 more frequently cause unfavorable drug-drug interactions, as the inactivated CYP3A4 has to be replaced by newly synthesized CYP3A4 protein. Most CYP3A4 inactivators are also PgP substrates/inhibitors, confounding the in vitro in vivo extrapolation. Clinicians should have good knowledge on these CYP3A4 inactivators and avoid their combination use. PMID- 15544436 TI - Human hepatocytes in primary culture: the choice to investigate drug metabolism in man. AB - Different types of hepatic tissue, including whole or split livers from organ donors or waste liver from therapeutic liver resections, are used to prepare human hepatocyte cultures. Characteristics of liver samples from different origins (gender, age, healthy/pathological status, xenobiotic treatment) as sources of human hepatocytes are key factors which notably determine viability and functionality of hepatocytes. The characterisation of the CYP system can be assessed in terms of activity (using specific substrates/inhibitors), protein (antibody analysis) and molecular biology-based mRNA amplification techniques (PCR technology and DNA microarrays). It could reasonably be considered that human hepatocytes reflect the heterogeneity of CYP expression in human liver and is a suitable model for drug metabolism studies. Several key issues need to be addressed at the early stages of drug development to better select drug candidates (metabolic profile and rate, identification of CYPs involved, drug drug interactions due to enzyme induction/inhibition). The metabolic stability and metabolite profile of new chemicals can be easily investigated by incubating the drugs with fully competent metabolic models like hepatocyte suspensions or 24 h-cultured hepatocytes. CYP inhibitory effects are usually screened in recombinant CYP enzymes or microsomes, however, the actual concentration of substrate and inhibitor available to the CYP enzyme depends on processes missing in subcellular models (transport mechanisms, cytosolic enzymes, binding to intracellular proteins). Since intact cells more closely reflect the environment to which drugs are exposed in the liver, cultured hepatocytes constitute a more predictive model for drug-drug interactions. Screening of CYP inducers cannot be done in microsomes as it requires a cellular system fully capable of expressing CYP genes. Primary hepatocytes are still the unique in vitro model for global examination of inductive potential of drugs (monitored as increases in mRNA content or activity). PMID- 15544444 TI - A perspective on the current strategies for the treatment of obesity. AB - The prevalence in obesity has increased dramatically over the past 30 years, more than double in the United States alone. Obesity is associated with an increased risk for type 2 diabetes mellitus, dyslipidemia, hypertension, biliary disease, obstructive sleep apnea, and certain types of cancer. The pathophysiology of obesity is complex, involving behavioral, environmental, and genetic factors. Current treatment options include behavior modification and lifestyle changes which incorporate weight-reducing diets and physical activity, FDA approved long term anti-obesity pharmacological agents sibutramine and orlistat, non-FDA approved over-the-counter (OTC) supplements and nutriceuticals, and, when appropriate, bariatric surgery. Without adequate prevention and treatment of obesity, government agencies have suggested that the direct and indirect costs associated with obesity may overwhelm the healthcare system. This brief review explores the current data available on treatments for the obese patient including the relative merits of different types of macronutrient composition (e.g., low carbohydrate vs. high carbohydrate diets) of weight-reducing diets, the value of resistance/ strength training in physical activity programs designed for the obese patient, the safety and efficacy associated with OTC supplements and nutriceuticals for weight reduction (e.g., Ephedra, conjugated linoleic acid (CLA), Garcinia cambogia/ hydroxycitric acid (HCA), chromium, pyruvate), the safety and efficacy of FDA-approved long-term obesity treatments sibutramine and orlistat, and bariatric surgery. PMID- 15544445 TI - Central G-Protein Coupled Receptors (GPCR)s as molecular targets for the treatment of obesity: assets, liabilities and development status. AB - In the last decade, the G-Protein-Coupled Receptor (GPCR) superfamily has emerged as a very promising and enriched source of therapeutic targets for the treatment of obesity. GPCRs represent the largest family of mammalian proteins, with approximately 1000 members. It is estimated that the GPCR family may comprise greater than 1% of the human genome and is the molecular target for approximately 30% of currently marketed drugs. Human GPCRs are modulated by a large variety of ligands, including peptides, lipids, neurotransmitters, nucleotides, ions and external sensory signals such as pheromones, tastes or odors. Many of the above ligands have been implicated in the physiological control of energy balance. This article will examine the biological rationale, assets, identified liabilities and current drug development status of these receptors as anti-obesity drug targets. PMID- 15544446 TI - Gut hormones as peripheral anti obesity targets. AB - Many peptides are synthesised and released from the gastrointestinal tract. Whilst their roles in regulation of gastrointestinal function have been known for some time, it is now evident that they also influence eating behaviour and thus potential anti obesity targets. Peptide YY (PYY) is released post prandially from the gastrointestinal L-cells with glucagon-like peptide 1 (GLP-1) and oxyntomodulin. Following peripheral administration of PYY 3-36, the circulating form of PYY, to mouse, rat or human there is marked inhibition of food intake. PYY 3-36 is thought to mediate its actions through the NPY Y2 GPCR. Obese subjects have lower basal fasting PYY levels and have a smaller post prandial rise. However, obesity does not appear to be associated with resistance to PYY (as it is with leptin) and exogenous infusion of PYY 3-36 results in a reduction in food intake by 30% in an obese group and 31% in a lean group. GLP-1 or oxyntomodulin, products of the prepreglucagon gene, decrease food intake when administered either peripherally or directly into the CNS. In addition, both have been shown to decrease food intake in humans. These effects are thought to be mediated by the GLP-1 receptor. Ghrelin, a huger hormone produced by the stomach, increases in the circulation following a period of fasting. Administration of ghrelin either peripherally or directly into the CNS increases food intake and chronic administration leads to obesity. Further infusion into normal healthy volunteers increases both food intake and appetite. Ghrelin is thought to act through the growth hormone secretagogue receptor (GHS-R). Obesity is the current major cause of premature death in the UK, killing almost 1000 people a week. Worldwide its prevalence is accelerating. The administration of the naturally occurring gut hormone may offer a long-term therapeutic approach to weight control. Here we consider the therapeutic potential of some gut hormones, and the GPCR's through which they act, in the treatment of obesity. PMID- 15544447 TI - Exercise in a pill: feasibility of energy expenditure targets. AB - The possibility of developing a pill to increase energy expenditure is explored by examining the metabolic processes involved. Such a pill should be targeted at organ systems involved in facultative thermogenesis. In rodents, these are brown adipose tissue (BAT) and skeletal muscle. Since BAT-mediated thermogenesis is not available in adult humans, emphasis here is on skeletal muscle. A hypothesis is presented based on three known facts: (1) plasticity of skeletal muscle, with interconversion of fiber types that differ in their fuel efficiency; (2) presence of thyroxine 5'-deiodinase type 2 (TD2) in human skeletal muscle; (3) gradual increase in thermogenesis that occurs during rehabilitation after starvation, probably in muscle. A low capacity thermogenic system, muscle efficiency thermogenesis (MET), is proposed to occur as adipose stores refill during the transition from famine to feasting to obesity. This system involves increased activity of TD2 and a T3-induced increase in proportion of type II fibers, less efficient at rest and during activity. The protective effect of this system is probably overwhelmed by long-term eating in excess of energy needs. Better understanding of the complex remodeling of differentiated muscle fibers in the conversions proposed and of the regulation of TD2 activity in human skeletal muscle may reveal targets for increasing energy expenditure in humans. In addition, the possibility of exploiting the plasticity of the adipose organ, with conversion of white adipocytes in white adipose tissue to atypical brown adipocytes and increasing thermogenesis in them is considered as another potential target for increasing energy expenditure in humans. PMID- 15544448 TI - Lipid metabolism and nutrient partitioning strategies. AB - The increasing prevalence of overweight and obesity worldwide is daunting and requires prompt attention by the affected, health care profession, government and the pharmaceutical industry. Because overweight/obesity are defined as an excess of adipose tissue mass, all approaches in prevention and treatment must consider redirecting lipid storage in adipose tissue to oxidative metabolism. Lipid partitioning is a complex process that involves interaction between fat and other macronutrients, particularly carbohydrate. In an isocaloric environment, when fat is stored carbohydrate is oxidized and vice versa. Processes that influence fat partitioning in a manner in which weight is maintained must be modified by changes in organ-specific fat transport and metabolism. When therapy is considered, however, changes in lipid partitioning alone will be ineffective unless a negative energy balance is also achieved, i.e. energy expenditure exceeds energy intake. The intent of this review is to focus on molecules including hormones, enzymes, cytokines, membrane transport proteins, and transcription factors directly involved in fat trafficking and partitioning that could be potential drug targets. Some examples of favorably altering body composition by systemic and/or tissue specific modification of these molecules have already been provided with gene knockout and/or transgenic approaches in mice. The translation of this science to humans remains a challenging task. PMID- 15544449 TI - Metabolic syndrome targets. AB - The metabolic syndrome is a cluster of easy-to-measure clinical phenotypes that serve as markers for increased risk for CVD and diabetes. There is no universal agreement as to the underlying pathophysiology of the metabolic syndrome. At its core, the metabolic syndrome is the result of energy excess; therefore treating obesity is a good strategy to reverse the clinical features of the metabolic syndrome. Hypertension is a special case, may not be part of the core pathophysiology of the metabolic syndrome, and will not be discussed. After a brief review of recent developments in the pathophysiology of the metabolic syndrome, this review will concentrate on peripheral targets in the following categories: ectopic fat and fat oxidation, intrinsic defects in substrate switching and mitochondrial biogenesis, lipolysis and lipid turnover, adipose tissue as an endocrine organ, nutrient / energy sensing systems, and inflammation. The advantages and pitfalls of these targets will be discussed with an eye towards the relevant literature. PMID- 15544450 TI - Immunization for persons infected with human immunodeficiency virus. AB - Immunization is an important measure to protect HIV-infected children and adults against certain vaccine preventable diseases. However, the antibody response, which is associated with the level of CD4+ T cell count, is frequently impaired in this group of patients. Certain vaccines enhance virus replication and transiently increase HIV viral load. Theoretically, vaccination should be given before the immune status of the patients is suppressed. Inactivated vaccines are generally safe and are beneficial for HIV-infected patients. These vaccines should be administered at appropriate age recommended for immunocompetent individuals. Live vaccines should be used with caution since some of the vaccines may be harmful to patients with severe immunologic suppression. Recommendations for immunization in HIV-infected patients may differ from country to country, depending on the availability and affordability of each vaccine, and the prevalence of each preventable disease. Vaccine trial in HIV-infected patients is needed in order to establish the most appropriate vaccine recommendation for this group of patients. PMID- 15544451 TI - Prevalence of toxoplasma encephalitis in AIDS patients treated with Didanosine hospitalised in a French infectious service. AB - In a previous work, we have showed in mice infected with an avirulent strain of Toxoplasma gondii and receiving a didanosine treatment, an important decrease of brain cysts. It is why, the purpose of this study was to investigate the effect of didanosine treatment on AIDS patients having developed Toxoplasma encephalitis. 60 patient reports were analyzed: 22 patients (group 1) did not received didanosine in their antiretroviral treatment and 38 (group 2) were treated with didanosine. The results showed that an antiretroviral therapy was prescribed for 93% of patients, 50% of them received only zidovudine and protease inhibitors were prescribed for 37%. The regimens given most frequently were those including zidovudine plus lamivudine or zidovudine plus indinavir. Among the group 1, 18% have had a relapse of Toxoplasma encephalitis. In the group 2, 37% of the patients suffered from one episode of TE while 16% have had two TE after the pause in their didanosine treatment, the maximum occurring between 4 and 24 months after the pause of didanosine. This study showed that didanosine seems to have an effect on cerebral cysts. Also, this work made a synthesis about the different treatment used in AIDS patients and the new molecules yet in development against T. gondii. PMID- 15544452 TI - Improving HIV infection management using antiretroviral plasma drug levels monitoring: a clinician's point of view. AB - Due to genetic and environmental factors, there are wide inter-patient differences when measuring drug exposure to a standard dose. If there is a relationship between drug exposure and efficacy or toxicity, this inter-patient variability carries various risks to develop toxicity or failure. Therapeutic drug monitoring is an attempt to adjust the dose to obtain a level within a therapeutic range consisting in a minimum plasma concentration needed to be efficacious and a maximum plasma concentration not to exceed to avoid toxicity. Many studies have shown a relationship between various pharmacokinetic parameters and drug toxicity or efficacy for HIV protease inhibitors (PIs) and non nucleoside reverse transcriptase inhibitors (NNRTIs). Therapeutic drug monitoring (TDM) proves to be a useful tool to assess adherence, to investigate drug-drug interactions between antiretroviral (ARV) drugs or with co-medications, to prevent some ARV drug toxicities, to adjust the dosage in particular populations, and to increase ARV efficacy of some drugs in naive patients. The integration of virological and pharmacological parameters, using inhibitory quotients, looks promising to improve therapy in ARV-experienced patients. Effective and non-toxic target concentrations will be determined for all present and future antiretroviral drugs covering the extended spectrum of naive patients to multiple failures. In this article, we review the rationale of TDM for antiretroviral drugs, the retrospective and prospective studies assessing plasma drug concentrations in relation with antiretroviral toxicity or efficacy, and the actually recommended or proposed indications for TDM. We also highlight the benefits and limits of this tool as an adjunct in the care of HIV-infected patients. PMID- 15544453 TI - Conformational changes in HIV-1 reverse transcriptase induced by nonnucleoside reverse transcriptase inhibitor binding. AB - Nonnucleoside reverse transcriptase inhibitors (NNRTI) are a group of small hydrophobic compounds with diverse structures that specifically inhibit HIV-1 reverse transcriptase (RT). NNRTIs interact with HIV-1 RT by binding to a single site on the p66 subunit of the p66/p51 heterodimeric enzyme, termed the NNRTI binding pocket (NNRTI-BP). This binding interaction results in both short-range and long-range distortions of RT structure. In this article, we review the structural, computational and experimental evidence of the NNRTI-induced conformational changes in HIV-1 RT and relate them to the mechanism by which these compounds inhibit HIV-1 reverse transcription. PMID- 15544454 TI - Interleukin-2 immunotherapy and AIDS-related cytomegalovirus retinitis. AB - Cytokines are small proteins produced by T lymphocytes that mediate immune responses. Those produced by the CD4+ Th1 subset induce cell-mediated immunity, whereas those produced by the CD4+ Th2 subset are more efficient at stimulating immunoglobulin production. The goal of cytokine immunotherapy is prevention or reduction of disease progression through stimulation of cell-mediated immunity (i.e., immune reconstitution) by administration of an exogenous Th1 cytokine such as interleukin-2 (IL-2). Cytokine immunotherapy has its origins in cancer immunobiology where IL-2 has been used successfully to manage several human cancers (metastatic melanoma, acute myelogenous leukemia, and metastatic renal cell carcinoma). More recent work has demonstrated cytokine immunotherapy to be effective at improving immune responses in patients with HIV-1 disease. To explore cytokine immunotherapy for sight-threatening AIDS-related human cytomegalovirus (HCMV) retinitis, we developed a mouse model of experimental murine cytomegalovirus (MCMV) retinitis that employs mice with MAIDS, a retrovirus-induced immunodeficiency syndrome. Systemic cytokine immunotherapy with IL-2, but not with interleukin-12 (IL-12), provides absolute protection against MAIDS-related MCMV retinitis by stimulation of the perforin-mediated pathway of cytotoxicity used by natural killer cells and cytotoxic CD8+ T cells to kill virus-infected cells. Our findings warrant additional studies on the use of cytokine immunotherapy for management of HCMV retinitis (and possibly other opportunistic infections) during HIV-1-induced immunodeficiency. We envision systemic cytokine immunotherapy as an altemative or adjunct to traditional antiviral chemotherapy for optimal management of AIDS-related HCMV retinitis. PMID- 15544455 TI - Neoplastic conditions in the context of HIV-1 infection. AB - HIV-1 infection predisposes to the development of specific types of cancer. Most cancers seen in the AIDS setting are related to oncogenic virus infections, such as Epstein-Barr virus (EBV), Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) and human papillomavirus (HPV). It is generally assumed that HIV-1 infection play a passive role in cancer development by impairing the host immune surveillance and increasing the risk of oncogenic virus infection. Recent insights, however, indicate that HIV-1 infection more actively promotes cancer growth. Experimental evidence has shown that HIV-1-encoded proteins can directly induce tumor angiogenesis and enhance KSHV transmission to target cells. Clinical evidence suggests that the oncogenicity of HPV is altered by the presence of HIV-1 infection irrespective of host immune status. The introduction of highly active antiretroviral therapy (HAART) has dramatically decreased the incidence of KS whereas the impact of HAART is variable in EBV-related lymphoma and HPV-related cervical cancer, suggesting that additional factors are involved in the pathogenesis of these cancers. Understanding the direct and indirect roles of HIV 1 in the pathogenesis of neoplastic conditions could provide the rationale for prevention and development of new treatments for AIDS-associated malignancies. PMID- 15544456 TI - Immunology of AIDS virus and mycobacterial co-infection. AB - While detrimental consequences of biological interaction between HIV and M. tuberculosis are well described, immune mechanisms underlying the evolution of HIV and mycobacterial coinfection remain poorly characterized. Recent studies in HIV-infected humans and SIV-infected monkeys have enhanced our understanding of complex issues of HIV and M. tuberculosis coinfection. Importantly, an anti-viral immune response and an associated control of AIDS virus infection can be induced during coinfection with mycobacterium or other pathogens in HIV-infected humans or SIV-infected monkeys. Restoration of anti-mycobacterial immunity during antiretroviral treatment can contribute to prevention and treatment of AIDS related mycobacterial diseases. This article discusses recent progress in immune aspects of HIV and mycobacterial coinfection. PMID- 15544457 TI - Recent advances in the development of HIV-1 Tat-based vaccines. AB - Over the last two decades most of the efforts in HIV vaccine development have been based on the use of the HIV Env with the goal to induce sterilizing immunity. However, as a result of Env variability disappointing results have been obtained in preclinical and phase III clinical trials. Although the objective of a preventive immunity still remains a priority, secondary endpoints (e.g. block of virus replication and disease onset) are being considered at the present as more achievable end-points in HIV vaccine development. This is based on accumulating evidence indicating that low viral load correlates with maintenance of immune functions and slow progression to disease, and that cell-mediated immunity plays a major protective role in the absence of sterilizing immunity. The promising results obtained in non-human primates with a vaccine based on a native Tat protein (B-clade), which is an early regulatory protein key for HIV replication and AIDS pathogenesis, highlights the importance of targeting the virus very early after infection. In particular, the immune response against Tat appears to modify the virus-host interactions at the very beginning of infection, thus containing the depletion of critical immune cells and the progression of infection. Moreover, since Tat targets and induces maturation of dendritic cells, has immunomodulatory activities and drives Th-1 and CTL responses, immunization with Tat may drive or increase these immune responses also against other HIV antigens to support an effective, long-lasting and hopefully even sterilizing antiviral immunity. Finally, Tat B-clade is similarly recognized by sera from individuals infected by different virus clades (A, B, C, D) supporting the concept of a cross-clade vaccine. Therefore, the Tat-vaccine should contain virus replication protecting from disease progression (non-sterilizing immunity) or even favoring an abortive infection. Although only a phase III clinical trial will establish the efficacy of this vaccine strategy, the Tat-vaccine has recently entered preventive and therapeutic phase I clinical testing in Italy to establish safety (primary-end-point) and immunogenicity (secondary end-point) and phase II studies are being prepared. PMID- 15544458 TI - Priority strategies for sustainable fight against HIV/AIDS in low-income countries. AB - Response to HIV/AIDS epidemic in resource-constrained countries is still woefully disappointing. This paper highlights some priorities shared at recent Florence World Conference (Florence, Italy: January 21st-24th, 2004) on how to overcome the obstacles still delaying sustainable fight against HIV/AIDS in developing world areas. Messages reported here result from selection made by the authors among challenging topics by more than one hundred speakers and have been chosen because of their value as most practical ways to secure prevention, treatment and care and achieve self-managing in fighting epidemic in income-limited settings. Building for success means to set up combined strategies--actively involving people living with HIV/AIDS (PLWHA) and grounded on coordination, coalition and partnership among all players--to prevent HIV transmission at mother-to-child, young and adult levels and to improve availability and access to laboratory testing and monitoring as well as to essential drugs for HIV/AIDS and related diseases. Building for success also means to provide women with reliable and affordable vaginal microbicides and to look for control of co-infections such as viral hepatitis, intestinal and sexually transmitted diseases as well as tuberculosis and malaria. Among the measures taken into account, the need for education and training is emphasised because its value may be even more important than funding in some countries. Priorities suggested in this paper reinforce each other underscoring the bidirectional value and synergy of the treatment and prevention strategies together with the need for keeping prevention in people giving successful antiretroviral treatment. In the Author's opinion, the current HIV/AIDS scenario may be reversed if the priorities taken into account will entirely be applied through adaptation to the different cultural backgrounds and social settings, and based on achievement of government's political will and accountability as well as on properly coordinated technical, financial and human support from international health cooperation. PMID- 15544459 TI - Immune modulation of HIV replication: relevance to HIV immuno- and neuro pathogenesis. AB - Much attention has been focused on CD4+ memory/primed T cells and monocytes/macrophages as the predominant targets of HIV infection. Considerable evidence, however, indicates that cell types that are classically not permissive to HIV infection, can be a potent source of productive virus replication in response to immune modulation. This immune modulation can be in the form of cytokine stimulation pre- or post-infection or by activating these "non conventional" targets. In this review, we will provide evidence for immune modulation of HIV infection with special emphasis on immune modulation of HIV replication in classically non-HIV permissive primary cells of both lymphoid and non-lymphoid origins. We will also examine the impact of HIV infection of these non-conventional targets as they pertain to both HIV immuno- and neuro pathogenesis. PMID- 15544460 TI - Application of combinatorial and parallel synthesis chemistry methodologies to antiparasitic drug discovery. AB - The discovery and development of novel drugs has been influenced over the last several decades by new techniques in medicinal chemistry. Combinatorial and parallel synthesis chemistry techniques have opened up immense opportunities in drug discovery and development efforts. These techniques, which include solid phase organic synthesis and polymer-assisted synthesis in solution, have been routinely applied to a number of therapeutic areas. Despite the flurry of activity that characterized small molecule drug discovery efforts in the early 1990s, it was only during the mid to late 1990s that combinatorial chemistry began to make an impact on antiparasite chemotherapy. This review focuses on the development and application of combinatorial and parallel synthesis methodologies to antiparasitic drug discovery from the mid 1990s to the end of 2002. Much of this work applies to small organic molecules as inhibitors of parasite targets although some of the early applications were to the synthesis of enzyme substrates. PMID- 15544461 TI - Dual inhibitors of the blood coagulation enzymes. AB - The search for an ideal anticoagulant has spanned decades and has resulted in several approaches and the identification of novel target molecules for preventing and treating thrombosis. The first group of new anticoagulant agents acting through direct inhibition of coagulation factors were inhibitors of thrombin, but factor Xa inhibitors and, most recently, factor VIIa inhibitors have become attractive candidates. The structures of thrombin, factor Xa and factor VIIa show similarities in their active sites and, for this reason, attempts have been made to develop synthetic agents containing in a single molecule inhibitory activity against two of the enzymes of the blood coagulation cascade. Such dual inhibitors are now in preclinical studies and are, potentially, new anticoagulant drugs with improved properties. The emphasis of this review will be placed on dual inhibitors of thrombin/factor Xa and factor Xa/factor VIIa. Comparison of the active sites of these enzymes is included for better understanding of the structural demands to be met in designing effective dual inhibitors. PMID- 15544462 TI - Improved, high yield synthesis of 3H-quinazolin-4-ones, the key intermediates of recently developed drugs. AB - Purine bases and their bioisosteric analogs are widely used as building blocks in combinatorial chemistry. Recently a great number of fused pyrimidine derivatives became known as potential drug molecules against various types of proliferative diseases, caused by over-expression of protein kinases. One of the most important compound families are quinazolines : e.g. the best inhibitor of EGFR tyrosine kinase is PD153035 (6,7-dimethoxy-4-(3'-bromophenyl)amino-quinazoline) and IRESSA (gefitinib, ZD1839), developed from this compound family, is presently the only one approved and granted drug by the FDA for the treatment of advanced non-small cell lung cancer (NSCLC). KF31327 (3-ethyl-8-[2-(4 hydroxymethylpiperidino)benzylamino]-2,3-dihydro-1H-imidazo[4,5-g]-quinazoline-2 thione dihydrochloride) from this group, showed significantly higher inhibitory activity on cyclic GMP-specific phosphodiesterase compared with those of sildenafil (Viagra). The synthetic procedures of the example compounds are based on imidoyl chloride intermediates that were prepared from the appropriate 3H quinazoline-4-ones. Although the key intermediates, quinazoline-4-ones, have been known since more than hundred years, their synthetic procedures have been improved much only in the past ten years. In this paper we reviewed the efficient synthetic methods of quinazolin-4-ones, and presented a novel, reliable method for their synthesis. There was no considerable effect of microwave-, or traditional thermal activation on the yield and compound purity. PMID- 15544463 TI - 6-oxo-morphinane oximes: pharmacology, chemistry and analytical application. AB - The permanent therapeutic importance of morphine derivatives in pain treatment has inspired continual synthetic efforts to modify the rigid pentacyclic systems in search for new selective analgesic agents. As a result, several morphinane oximes have been synthesized recently, which have the additional advantage of possessing an oxime group that can provide a method for selective determination of opiate alkaloids in biological matrices. The oximes of hydrocodone and oxycodone have stronger analgesic effect than the parent ketones and they also proved to be effective in preventing the respiratory depressant and hypotensive actions of fentanyl. In this work a review is given on the present status of oxime pharmacology, chemistry and analysis and also the oxime and O-methyl oxime formation of 6-oxo-morphinanes with therapeutic interest (codeinone, oxycodone, hydrocodone and 14-OH-codeinone). The oxime formation was monitored by reversed phase HPLC and the chromatographic properties of oxime isomers have been characterized. The assignation of the individual isomers isolated by preparative HPLC was performed by (1)H NMR spectroscopy based on the chemical shift differences of the 5-H signals. In this way the isomeric ratio in the oxime products could also be determined. It was found that in the case of Delta(7)-6 oxo-morphinanes, depending on the substituents, the formation of the Z-isomer highly dominates (73-96%) over that of the E-isomer. However, for the saturated 7,8-(dihydro) derivatives the E-isomer is definitely preferred (>98%). In conclusion of a survey on the theoretical background of oxime isomerism, the conformational differences between the saturated and unsaturated morphinane systems were found responsible for the different E/Z ratios. On the basis of the isomeric ratio and the on-line CD and UV spectra of the pure isomers, the molar ellipticities and absorbancies of the isomers were calculated by a parameter estimation method. PMID- 15544464 TI - Structural aspects of carbohydrates and the relation with their biological properties. AB - Structural diversity of carbohydrates plays a crucial role in their large variety of roles in biological systems. This paper focuses on aspects of structure and biological functions of three classes of carbohydrates, N-linked oligosaccharides, blood group oligosaccharides and glycosaminoglycans. Conformations and dynamics in solution, as well as structure of protein carbohydrate complexes are discussed. A short overview also describes theoretical and experimental methodologies that are used in current glycobiological research, particularly high-resolution NMR spectroscopy, X-ray crystallography and methods of computational chemistry. PMID- 15544465 TI - Recent advances in 4'-thionucleosides as potential antiviral and antitumor agents. AB - The classical 4'-oxonucleoside analogs exhibit interesting biological activities such as antibiotic, antiviral and antitumor, which are believed to be the result of inhibition of the viral or cellular DNA or RNA polymerase after being converted to their corresponding 5'-triphosphates. However, the activity of 4' oxonucleosides were limited by their susceptibility to degradation by nucleoside phosphorylases or acid hydrolysis. This aspect called for the chemical modification of the carbohydrate portion. This compulsion led to two kinds of strategies; (1) replacement of the 4'-oxygen by the methylene group - carbocyclic nucleosides; (2) replacement of the 4' oxygen by sulphur-4'-thionucleosides. This group has also conferred the resistance to the nucleoside cleavage. Although, there were some pioneering work on 4'-thionucleosides in 1960s and 1970s, the interest in this group of compounds was rekindled by the antiviral activities of 2'-deoxy-4'-thionucleosides reported independently by Secrist et al. and Walker et al. Subsequent contributions by the other authors, enhanced its standing as an important class of antiviral agents. Following is a reasonably exhaustive account of this class of compounds reported after 1990. PMID- 15544466 TI - 6-phosphogluconate dehydrogenase: a target for drugs in African trypanosomes. AB - New drugs are urgently required for Human African Trypanosomiasis (sleeping sickness), a disease which has re-emerged as a major health threat in Sub-Saharan Africa. The third enzyme of the pentose phosphate pathway, 6-phosphogluconate dehydrogenase, has been shown to be a good target for drugs. The enzyme is essential to the trypanosomes that causes sleeping sickness and structural differences when compared to its mammalian counterpart allow for selective inhibition. Three series of inhibitors have been designed, these include phosphorylated carbohydrate substrate and transition state analogues, non carbohydrate substrate analogues and also triphenylmethane-based compounds. All have shown selective inhibition of the trypanosomal 6-phosphogluconate dehydrogenase and representatives of each have trypanocidal activity. PMID- 15544467 TI - Biosynthesis of peptide hormones derived from precursor sequences. AB - The release of hormones is subject to a complex and finely tuned regulation system. The biosynthesis plays a key role by specifically converting the prohormone precursor into its biological active product(s). A family of mammalian proteases could be identified to be responsible for the endoproteolytic processing. These subtilisin/kexin-like prohormone convertases (PC) recognize their substrates at single or pairs of basic residues with a high substrate specificity. The so far known seven members include PC1/3, PC2, furin/PACE, PACE4, PC4, PC5/6 and PC7/SPC7/LPC/PC8. PC1/3 and PC2 are the most important enzymes for the processing of prohormones, whereas furin is the only one that causes lethality in knock-out models. Tissue-specific co-localization of the prohormone and the PC as well as distinct characteristics of both, like the secondary structures, determine the possible conversion processes. Identification of such determinants implies a great potential for the development of novel drug targets. To obtain sufficient amounts for the in vitro characterization of prohormones, chemical and recombinant synthesis methods have been developed. Application of expressed protein ligation lead to the semisynthesis of the first chemically modified analogs of a full-length proneurohormone (pro-neuropeptide Y). Structural analyses mainly on peptides of the pro-oxytocin/neurophysin system and on prosomatostatin highlighted the importance of flexible turn or loop structures adjacent to the cleavage site for the specific substrate-enzyme active site interaction. Prohormones and their processing show multiple functions. Therapeutic application including PC inhibitors is very promising for the treatment of disorders like cancer. PMID- 15544468 TI - Influence of mutations in the hepatitis B virus genome on virus replication and drug resistance--implications for novel antiviral strategies. AB - Persistent infection with the hepatitis B virus (HBV) represents a major health problem worldwide with over 350 million patients at risk of developing liver cirrhosis or hepatocellular carcinoma. HBV is a small, partially double-stranded DNA virus with four overlapping genes and a unique life cycle, creating an intracellular pool of covalently closed circular DNA molecules for persistence and an RNA template for replication via reverse transcription. Mutations occur frequently, and particular selection pressures, both endogenous (host immune clearance) and exogenous (vaccines and antivirals), readily select escape mutants. For example, HBeAg-negative chronic HBV infection with either basal core promoter or precore mutations is predominant in many parts of the world. Therapy of HBV infection with the nucleoside analogue lamivudine frequently leads to the selection of drug-resistant strains with polymerase mutations. Treatment options for chronic HBV infection include at present either interferon-alpha or the oral nucleos(t)ide analogues lamivudine or adefovir. However, these drugs have drawbacks, including possible serious side effects and low response rates in HBeAg-negative patients in the case of interferon or recurrence of viremia after cessation of therapy and development of escape mutants after a long period of treatment with nucleoside inhibitors. Recent advances of in vitro and in vivo models allow to study new antiviral strategies, including novel nucleoside analogues, nucleocapsid inhibitors or small interfering RNA. This review summarises the impact of clinically relevant mutations in the HBV genome on viral replication and drug sensitivity, the current status of therapy and promising future perspectives on novel drug regimens. PMID- 15544469 TI - S-nitrosothiols as nitric oxide-donors: chemistry, biology and possible future therapeutic applications. AB - In recent years, the gaseous molecule nitric oxide (NO) has been shown to be involved in many important biological events. S-Nitrosothiols are biological substances derived endogenously from NO, and are found in a variety of tissues exhibiting NO-mimetic activity. Fundamental studies on the chemical aspects of S nitrosothiols have become an integral part of NO research, with a view to further understanding the numerous biological functions both of NO and of S-nitrosothiols themselves. It has often been suggested that S-nitrosothiols represent a means either for the storage or transport of NO, although the evidence for this is sparse. Many S-nitrosothiols have now been synthesized chemically, and at present they show the most promise as clinically useful NO-donor drugs. In this review, we examine in detail the biological functions of S-nitrosothiols, as well as the chemical properties and biomedical applications of these compounds. PMID- 15544470 TI - Torsadogenic cardiotoxicity of antipsychotic drugs: a structural feature, potentially involved in the interaction with cardiac HERG potassium channels. AB - Many non-cardiovascular drugs of common clinical use cause, as an unwanted accessory property, the prolongation of the cardiac repolarisation process, due to the block of the HERG (Human Ether-a-go-go Related Gene) potassium channel, responsible for the repolarising I(Kr) current. This delayed cardiac repolarisation process can be often unmasked by a prolongation of the QT interval of the ECG. In these conditions, premature action potentials can generate morphologically anomalous after-polarisations, and trigger a dangerous kind of polymorphic ventricular tachyarrhythmia, known as torsade de pointes, which can evolve in ventricular fibrillation and death. The risk associated with the torsadogenic cardiotoxicity of drugs, which prolong the QT interval has been the topic of documents produced by many health authorities, giving important issues about the preclinical and clinical evaluation of cardiac safety. Besides, public and private research laboratories developed several experimental in vitro or in vivo strategies, aimed to an early recognition of the influence of a drug (or of a drug-candidate) on the HERG channel and/or on the cardiac repolarisation process. Also the identification of a possible pharmacophore model, common in all or at least in numerous torsadogenic drugs, could represent a first step for the development of useful in silico approaches, allowing a preliminary indication about the potential torsadogenic property of a given molecule. In this work, we described the electrophysiological basis of torsade de pointes and listed several pharmacological classes of torsadogenic drugs. Among them, we focused our attention on antipsychotics, with an accurate overview on the experimental and clinical reports about their torsadogenic properties. Moreover, a common structural feature exhibited by these drugs, despite of their remarkable chemical differences, is evidenced by a computational approach and is indicated as a possible "facilitating" requirement for their torsadogenic properties. Together with other remarks, coming from different computational studies, the individuation of a satisfactory "toxicophore" model could be greatly useful, for the theoretical prediction of torsadogenic properties of a given chemical moiety and for the design of new drugs devoid of such an undesired and potentially lethal side-effect. PMID- 15544471 TI - Intracellular location of KATP channels and sulphonylurea receptors in the pancreatic beta-cell: new targets for oral antidiabetic agents. AB - Diabetes Mellitus is by far one of the most propagated chronic diseases, affecting 150 million people worldwide. This affliction is caused by a malfunction of pancreatic endocrine cells, which provokes a failure in the insulin release and glucose homeostasis. Plasma membrane K(ATP) channels have a key role in the stimulus-secretion coupling of pancreatic beta-cells. Consequently, many investigations have developed efficient drugs for the treatment of diabetes, such as sulphonylureas, which specifically close K(ATP) channels leading to an enhanced insulin secretion. Recent studies show that, in addition to its well-known plasma membrane location, sulphonylurea receptors and sulphonylurea-sensitive K(ATP) channels are also present in various intracellular sites including secretory granules, mitochondria, endoplasmic reticulum and more recently, the nucleus. What roles do they play in these organelles? Intracellular K(ATP) channels and sulphonylurea receptors, which operate in conjunction with classical pathways, can provide specific signaling circuits to establish direct links between extracellular signals and different cell functions, such as secretion or gene expression. The study of these intracellular channels provides novel perspectives in the signal transduction of the pancreatic beta-cell, and may offer clues for the development of new strategies in diabetes therapy. In this review we will address this topic with special emphasis on the biophysical basis and functional implications in the pancreatic beta-cell. PMID- 15544472 TI - Glucose transporter and Na+/glucose cotransporter as molecular targets of anti diabetic drugs. AB - Glucose transporters, or membrane proteins, which incorporate glucose into the cell, can be divided into two groups: the facilitative type glucose transporter (GLUT), and the sodium/glucose cotransporter (SGLT). Among the GLUT family isoforms, GLUT4 is particularly important for maintaining glucose metabolism homeostasis since it is involved in insulin or exercise-induced glucose transport into muscle and adipose tissues via movement from intracellular sites to the plasma membrane in response to stimulation. Thus, agents which induce GLUT4 translocation or improve insulin sensitivity, involved in this insulin-induced step, hold the promise of being potent anti-diabetic drugs. On the other hand, SGLT is expressed specifically in the intestines and kidneys. Oral administration of a SGLT inhibitor, T-1095, lowers the blood glucose concentration via excretion of glucose in the urine, due to suppression of renal SGLT function. In addition to this direct blood glucose lowering effect, T-1095 has been shown to restore impaired insulin secretion from pancreatic beta-cells, as well as to improve insulin resistance in muscle and liver. Thus, this SGLT inhibitor is regarded as a novel and promising agent for the treatment of diabetes mellitus. PMID- 15544473 TI - Inhibitors for proteins endowed with catalytic and non-catalytic activity which recognize pTyr. AB - Reversible phosphorylation of Tyr residues in proteins plays a central role in the transduction of signals. For both SH2 domains and for protein tyrosine phosphatases (PTPs) the phosphate group of phosphotyrosine (pTyr) of peptides provides a key affinity element, but its highly charged nature and its hydrolytic lability render it unsuitable in inhibitor design. The research in the recent years has been addressed to find pTyr bioisosters devoid of the phenylphosphate moiety and more potent inhibitors with less peptidic character. Several derivatives were prepared as pTyr bioisosters, and their activity appears to depend on the nature of the substrate, peptidic or low-molecular weight compounds, in which they are placed. In the field of PTPs, the research was mainly focused on new and selective PTP1B inhibitors, possibly useful in the treatment of Type 2 diabetes. The discovery of non-peptidic low molecular weight compounds able to inhibit PTP1B, by means of docking procedures and HTS screening, and the presence of secondary binding sites on PTP1B afforded new potent and selective inhibitors; several leads devoid of negative charges were also found. To date, however, few compounds have been tested In vivo and found to show a significant activity in diabetic mouse models. Other neutral compounds, mainly quinones, were found to inhibit CD45 and Cdc25. Several papers have appeared in recent years on the discovery of new Grb2, Src, Syk, and Lck SH2 domains binding antagonists. In this field very good inhibitors derived from high affinity peptides were found, with less peptidic character and with a reduced number of negative charges; however the presence of some negative charges, especially the one present on the pTyr bioisoster moiety, seems to be indispensable. As regards Grb2, Src and Lck SH2 domains, rigidification of the starting high affinity binding peptides afforded derivatives with improved affinity; cellular activity was achieved by modification of the side chains of these inhibitors. PMID- 15544474 TI - 5-(1-Substituted) alkyl pyrimidine nucleosides as antiviral (herpes) agents. AB - The treatment of viral diseases remains one of the major challenges to modern medicine. During the past two decades there has been increased recognition of the consequences of serious viral illnesses that are not controlled by vaccination. These illnesses include human immunodeficiency virus, human herpes viruses, and viruses that cause hepatitis. There are now eight pathogens recognized in the herpes virus family that cause infections in humans. Infections by the herpes viruses are opportunistic and often life-threatening, leading to significant morbidity and mortality in the increasing number of chronically immune compromised individuals such as AIDS patients, cancer patients and transplant recipients on immunosuppressive therapy. Nearly all individuals with AIDS are infected with one or more of the herpes viruses. Antiviral therapy with guanosine nucleoside analogs acyclovir and ganciclovir has had a major impact on diseases caused by herpes simplex virus type-1 and type-2 (HSV-1, HSV-2), Varicella zoster virus (VZV), and human cytomegalovirus (HCMV) but development of resistant virus strains and the absence of any effective treatment for other members of the herpes family provide a stimulus for increased search of new agents effective against various herpes viruses. Pyrimidine nucleosides have taken up an important role in the therapy of virus infection. Significant progress in the study of anti herpes nucleosides has been made by the advent of 5-substituted pyrimidine nucleosides such as 5-iodo-, 5-ethyl-, 5-(2-chloroethyl)-, and (E)-5-(2 bromovinyl)- derivatives of 2'-deoxyuridine. These are highly specific inhibitors of HSV-1, HSV-2, and/or VZV infections. However, Epstein Barr virus (EBV) and HCMV are much less sensitive to these agents. In 5-substituted pyrimidine nucleosides the nature of substituents, particularly at the C-5 position, has been found to be an important determinant of anti-herpes activity. Structural requirements at the C-2 carbon of the 5-substituent of pyrimidine nucleosides have been well established for anti-herpes activity. However, there is little qualitative or mechanistic knowledge of the derivatives with substitution at the C-1 carbon of the 5-substituent of pyrimidine nucleosides. During the last few years of our research, we have investigated a variety of C-1 functionalized substituents at the 5-position of the pyrimidine nucleosides to determine their usefulness as antiviral (herpes) agents. In the 5-(1-substituted) group of pyrimidine nucleosides, we demonstrated that novel substituents present at the C 1 carbon of the 5-side chain of the pyrimidine nucleosides are important determinants of potent and broad spectrum antiviral (herpes) activity including EBV and HCMV. In this article the work on design, synthesis and structure activity relationships of several 5-[(1-substituted) alkyl (or vinyl)] pyrimidine nucleoside derivatives as potential inhibitors of herpes viruses is reviewed. PMID- 15544475 TI - The clinical relevance of advanced glycation endproducts (AGE) and recent developments in pharmaceutics to reduce AGE accumulation. AB - Advanced glycation endproducts (AGE) are a class of compounds resulting from glycation and oxidation of proteins, lipids or nucleic acids. Glycation is the non-enzymatic addition or insertion of saccharide derivatives to these molecules. This leads to the formation of intermediary Schiff bases and Amadori products and finally to irreversible AGE. This classical view has been modified in recent years with recognition of the importance of oxidative and carbonyl stress in endogenous AGE formation. AGE may also have exogenous sources, in certain foods and tobacco smoke. A whole class of specific and non-specific receptors binding AGE has been characterized. Apart from cross-linking of proteins by AGE, the effects of receptor stimulation contribute to the development of chronic complications of conditions like diabetes mellitus, renal failure, and atherosclerosis. Possible interventions to reduce the effects of AGE accumulation include AGE formation inhibitors or breakers, or receptor blockers, but possibly also dietary interventions. Some of the problems with current assay or diagnostic techniques, and several unresolved issues on the role of AGE in disease will be discussed. Our review will focus on the clinical and pharmaceutical implications of these developments. PMID- 15544476 TI - Exploring Ramachandran and chi space: conformationally constrained amino acids and peptides in the design of bioactive polypeptide ligands. AB - Ligand binding and concomitant changes in receptor structure provide the means to target signal transduction pathways. With appropriate refinement of the ligand's interaction with the "receptor," one in theory could produce ligands that have greater therapeutic benefits. This review will discuss how, when these ligands are amino acids and peptides, the introduction of appropriate conformational constraints provides a powerful strategy for improved drug design. This review will discuss how various constraints on amino acids can provide a powerful tool for ligand design, determination of the three dimensional pharmacophore and new insights into receptor systems and information transduction. Through the use of constrained ligands, new information regarding their interaction with their "receptor" systems, and further refinement of the use of constraints, scientists can produce more beneficial drugs for mankind. PMID- 15544477 TI - N- and Calpha-methylation in biologically active peptides: synthesis, structural and functional aspects. AB - Numerous backbone constraints can be used to develop pseudopeptides or pseudomimetics of biologically active peptides. Among those, N- and Calpha-methyl amino acids that can be incorporated by solid-phase peptide synthesis in a bioactive sequence represent important tools to restrict phi and psi angles of peptide backbone. This review will focus on the chemical syntheses of N- and Calpha-methyl amino acids, their effects on peptide conformation and structure, and their role on the peptide stability towards enzymatic degradation and on the biological activities of the resulting analogues. PMID- 15544478 TI - Development of conformationally restricted analogues of bradykinin and somatostatin using constrained amino acids and different types of cyclization. AB - The structure-based design of peptide drugs requires the knowledge of the bioactive conformation. Studies on this receptor-bound 3D structure require linear or cyclic analogues with strongly reduced flexibility, but high biological activity, since only analogues with retained potency have preserved the bioactive conformation. Constrained amino acids containing double bonds or bulky substituents at the N(alpha)-, C(alpha)- and C(beta)-atom as well as at the aromatic ring atom were successfully applied to obtain potent and stable analogues of bradykinin and somatostatin, which due to their restricted conformation were suitable objects for conformational studies. Besides the generation of constrained cyclic analogues with improved biological and pharmacological properties, cyclic peptides were used as convenient models for the study of turn formations. Cyclization of the linear peptide bradykinin was performed by linking the N-terminus and the C-terminus, and in both bradykinin and somatostatin by cyclization using the amino acid side chains and by backbone cyclization. The later requires the introduction of N(alpha)-functionalised amino acids for ring closure which can be performed either through incorporation of N(alpha)-functionalised amino acids or dipeptide building units. Conformational analysis of a cyclic bradykinin analogue by means of NMR-studies together with molecular dynamics simulation led to a quasicyclic 3D structure with two turns and together with other 3D structures provided a pharmacophore model of bradykinin antagonists. PMID- 15544479 TI - Contribution of conformationally constrained calcitonin (Ct) analogs to the understanding of the structural and conformational requirements of calcitonin bioactivity and to the design of potent agonists. AB - Restricting the conformational flexibility of medium-sized linear polypeptides is a valuable approach to identify and characterize the structural and conformational features that define their biological activities and to design analogs with enhanced agonistic or antagonistic properties and with potential therapeutic applications. The calcium-regulating and bone resorption-inhibiting hormone calcitonin (Ct) is a conformationally flexible polypeptide of 32 amino acid residues that has long been applied therapeutically for the treatment of osteoporosis and other bone disorder diseases. This review describes studies on the structural and conformational features of the Ct sequence that are relevant for Ct bioactivity and focuses on research work performed on rationally designed conformationally constrained Ct analogs as tools for the understanding of the molecular basis of Ct bioactivity and as potential candidates or lead structures for novel Ct-based bone disorder therapeutics. PMID- 15544480 TI - Constrained analogs of osteogenic peptides. AB - Osteogenic peptides are, or have potential to be, therapies for the treatment of osteoporosis, fracture repair, and repair of loosened bone implants. Human parathyroid hormone has been approved for the treatment of post-menopausal osteoporosis. Constrained analogs of PTH and the parathyroid-hormone related peptide (PTHrP) have aided the understanding of how PTH and PTHrP bind to their common receptor and some of these analogs have improved properties that make them possible candidates for clinical trial. Cyclization by lactam formation has shown that a core region of human PTH (hPTH) from residues 16-26 binds as an alpha helix to the receptor and that the biological effects are remarkably sensitive to ring size. Appropriate cyclization in this region of the molecule not only has yielded analogs with improved receptor activation but also ones less susceptible to protease degradation and thus more active in vivo. Cyclization has been less successful in the N-terminus region, residues 1-12, of hPTH(1-34) with only a cyclization between residues 6 and 10 showing some promise. The growing understanding of how this region binds to the receptor will lead to other productive constraints. This review also covers the potential of a different class of molecule, the osteogenic growth peptide (OPG), as an anabolic bone agent. These molecules have much weaker anabolic effects than PTH and cyclization does not result in improved activity. However, the information gained from these studies may yield analogs with better pharmacological profiles. PMID- 15544481 TI - Therapeutic potential of natural compounds that regulate the activity of protein kinase C. AB - Protein kinase C (PKC) is a family of serine/threonine kinases that regulates a variety of cell functions including proliferation, gene expression, cell cycle, differentiation, cytoskeletal organization, cell migration, and apoptosis. The PKC signal transduction cascade coordinates complex physiological events including normal tissue function and repair. Disruption of the cellular environment through genetic mutation, disease, injury, or exposure to pro oxidants, alcohol, or other insults can induce pathological PKC activation. Aberrant PKC activation can lead to diseases of cellular dysregulation such as cancer and diabetes. Can aberrant activation of PKC be reversed? Even 25 years after the identification of PKC, therapeutic regulation of PKC activity remains an emerging field. Because the function of each isoform remains to be elucidated, isoform specific control of gene expression is a current challenge. Natural compounds are important regulators of PKC activity, with both preventive and therapeutic efficacy. Antioxidants including vitamin A (retinoids), vitamin C (ascorbic acid) and vitamin E (tocopherols) show promise for reversal of PKC activation. beta-carotene and retinoids function as anticarcinogenic agents and antagonize the biological effects of pro-oxidants on PKC. Vitamin E reverses the deleterious effects of hyperglycemia and diabetes by down-regulating PKC activity. Antioxidants in red wine provide cardioprotective effects. However, alcohol consumption also induces oxidative stress and disrupts PKC and retinoid function in the fetus and the adult. This review examines modulation of PKC activity by natural compounds and pharmacologic analogues which can be used effectively to prevent or treat common diseases associated with aberrant activation of PKC. PMID- 15544482 TI - Haemoglobinopathy in pregnancy: diagnosis and treatment. AB - Haemoglobinopathies differ in geographic prevalence but together are amongst the most common genetic disorders worldwide. Despite huge diagnostic progress, therapeutic options remain limited, with many treatments still at the experimental stage, no more so than in pregnancy: not only does the presence of a fetus subject treatments to greater limitations, but also any worsening of the anaemia as pregnancy progresses results in higher fetomaternal morbidity and mortality. Anaemia weakens the response to peripartum blood loss, with the risk of postpartum complications. Until recently the standard conventional therapy for severe anaemia was (repeated) blood transfusion, with its well-known risks. Recombinant human erythropoietin (rhEPO) can induce fetal haemoglobin and is a safer, if less immediately effective, alternative for the correction of anaemia in pregnant patients with haemoglobinopathy. PMID- 15544483 TI - The design, structure, and clinical update of small molecular weight matrix metalloproteinase inhibitors. AB - Matrix metalloproteinases (MMPs) are a family of zinc-containing enzymes involved in the degradation and remodeling of extracellular matrix proteins. Under normal physiological conditions, the activities of these enzymes are well-regulated by endogenous tissue inhibitors of metalloproteinases (TIMPs). Chronic stimulation of MMP activities due to an imbalance in the levels of MMPs and TIMPs has been implicated in the pathogenesis of a variety of diseases such as cancer, osteoarthritis, and rheumatoid arthritis. Thus, MMP inhibitors are expected to be useful for the treatment of these disorders. Because of their importance in a variety of pathological conditions, a number of small molecular weight MMP inhibitors have entered clinical trials in humans. However, the results of these trials have been extremely disappointing and have led many investigators to conclude that MMP inhibitors have no therapeutic benefit in human cancer. To date, the first generation MMP inhibitors exhibited poor bioavailability while second-generation compounds revealed that prolonged treatment caused musculoskeletal pain and inflammation or had a lack of efficacy. This article describes the design of small molecular weight MMP inhibitors, a brief description of available three-dimensional MMP structures, a review of the proposed therapeutic utility of MMP inhibitors, and a clinical update of compounds that have entered clinical trials in humans. The experimentally determined structures used in the structure-based design of MMP inhibitors are thoroughly covered. Major emphasis is on recently published and/or patented potent MMP inhibitors, from approximately January 2000 to April 2003, and their pharmacological properties. Protein inhibitors of these proteolytic enzymes, i.e. TIMPs, will not be discussed. PMID- 15544484 TI - RNase P: a promising molecular target for the development of new drugs. AB - RNase P is an ubiquitous and essential endonuclease in tRNA biogenesis, which generates the mature 5'-termini of tRNAs. RNase P activities have been identified in all three kingdoms of life (Bacteria, Archaea, Eukarya). Most forms of RNase P are ribonucleoproteins, i.e., they consist of an essential RNA and protein subunits. In bacteria and in some archaea, the catalytic function of this enzyme resides entirely in its RNA subunit, which is one of firstly identified ribozymes. Its high structural and functional diversity among representatives of a vast variety of phylogenetic domains indicates that RNase P could also serve as a molecular target of and a useful screening system for the biological activity of different compounds and give more insight into the molecular mechanisms of their action inside the cell. The emerged information from recent studies on the mechanism and structural idiosyncrasies of RNase P provides a convenient platform for designing specific inhibitors for this ribozyme and potential areas of its application in gene therapy. This review summarises the current information on the effect of several protein synthesis inhibitors, retinoids and arotinoids, vitamin D analogues and anthalin on the activity of RNase P. PMID- 15544485 TI - Pharmacophore modeling and three dimensional database searching for drug design using catalyst: recent advances. AB - Perceiving a pharmacophore is the first essential step towards understanding the interaction between a receptor and a ligand. Once a pharmacophore is established, a beneficial use of it is 3D database searching to retrieve novel compounds that would match the pharmacophore. As the 3D searching technology has evolved over the years, it has been effectively used for lead optimization, combinatorial library focusing, as well as virtual high-throughput screening. This paper is an update to the original paper published in this journal earlier: Kurogi, Y, and Guner, O. F. "Pharmacophore Modeling and Three-Dimensional Database Searching for Drug Design Using Catalyst," in Current Medicinal Chemistry, 2001, 8(9), 1035 1055. PMID- 15544486 TI - Pharmacokinetic distribution of 67Cu(II)2[3,5-diisopropyl(carboxy 14C)salicylate]4 among murine tissues. AB - Non-toxic doses of tetrakis-mu-3,5-diisopropylsalicylatodicopper(II) [Cu(II)2(3,5 DIPS)4] have been found to have anti-inflammatory, analgesic, anti-ulcer, anti colitis, anti-convulsant, anti-cancer, anti-mutagenic, anti-carcinogenic, and anti-diabetic activities and, in addition, facilitates recovery from lethal irradiation and ischemia-reperfusion injuries. The goal of this research was to determine the time-dependent tissue distribution and persistence of 67Cu and the 14C labeled salicylate ligand, carboxy-14C-3, 5-diisopropylsalicylate [3,5 DIP(carboxy-14C)S], following subcutaneous administration of a 50 micromole per kilogram of body mass dose of double labeled tetrakis-mu-3,5-diisopropyl[carboxy 14C]salicylatodiaquo [67Cu]dicopper(II) 67Cu(II)4[3,5-DIP(carboxy-14C)S]4. This compound was administered to nine groups of six 20 gram female C57BL/6 mice and blood, liver, kidney, intestine, lung, thymus, femur, muscle, spleen, and brain tissues removed and analyzed for 67Cu and 14C at 0.5, 1, 3, 6, 12, 24, 48, 72, and 96 hours after treatment. These data were then analyzed using a pharmacokinetic model simulation program. Both 67Cu and 14C were found in all tissues as well as urine and feces at 0.5 hour after administration. As anticipated, 67Cu entered the liver storage pool; it was conserved by the kidneys, and subsequently underwent release in maintaining 67Cu levels in all other tissues. While the presence of 67Cu correlated with the presence of the salicylate ligand, 3,5-DIP (carboxy-14C)S, early in the course of this experiment, the ligand was lost via ligand exchange and could not be measured in blood, kidney, intestine, lung, thymus, spleen, and brain after 24 hours following administration. However, 3,5-DIP(carboxy-14C)S persisted in liver, femur, and muscle throughout the 5-day period of study. It is suggested that marked lipophilicity accounts for its very rapid distribution to all tissues wherein it undergoes ligand exchange as 67Cu is incorporated into Cu-dependent enzymes and proteins and persists in tissues based upon physiological demand for Cu in meeting normal biochemical requirements. PMID- 15544487 TI - Non-peptidic GnRH receptor antagonists. AB - Gonadotropin-releasing hormone (GnRH) or luteinizing hormone-releasing hormone (LHRH) is a decapeptide (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH2) hypothalamic hormone that acts upon 7-trans membrane spanning GnRH receptors in the pituitary. This action leads to the secretion of the gonadotropins, luteinizing hormone (LH) and follicle-stimulating hormone (FSH) that in turn act on the reproductive organs regulating gonadal steroid production, spermatogenesis and follicular development. Peptidic agonists of the GnRH receptor have been known for many years and are currently employed therapeutically in the treatment of prostate and breast tumours, uterine fibroids, precocious puberty, endometriosis, premenstrual syndrome, contraception and infertility. Peptidic antagonists to date have only been employed commercially in the treatment of infertility during assisted reproductive therapy; however, many peptidic antagonists are currently in late stage development for many of the aforementioned indications. Whilst peptidic agonists and antagonists of the GnRH receptor have been discovered and exploited clinically, they are limited to predominantly parenteral administration due to their poor oral bioavailability. Recently, several small molecule GnRH antagonist series have been discovered offering the prospect of orally active therapeutics based on GnRH receptor antagonism. This article will review the current medicinal chemistry literature and structure activity relationships known for non-peptidic GnRH receptor antagonists. PMID- 15544488 TI - Zebrafish as a genomics research model. AB - The zebrafish (Danio rerio) is a recent addition to the genomic scientists' repertoire of vertebrate animal model systems. Unlike simple invertebrates such as the fly or the nematode, this teleost maintains the biological and genomic complexity found in higher vertebrates. Furthermore, the zebrafish has many advantageous technical and genomic properties that open the door to experimental approaches not practical using more classical models. The zebrafish genome can be functionally accessed using both forward and reverse genetics based approaches. A notable recent addition to the zebrafish genomics toolbox is the development of morpholino-based antisense gene inhibition for sequence-based 'knockdown' screening. This method offers the opportunity to examine the role of significant subsets of the vertebrate genome for specific gene function in vivo. The zebrafish embryo can rapidly provide critical information for drug target discovery purposes when examined with an emphasis on clinically-relevant biological processes. Finally, the advent of chemical genetics in zebrafish suggests that, in addition to the identification and understanding of drug targets and their biology, this system will be a powerful tool in the direct development of novel pharmaceuticals in the near future. PMID- 15544489 TI - Anti-genes: siRNA, ribozymes and antisense. AB - Scientists have been working on strategies to selectively turn off specific genes in diseased tissues for the past thirty years. In the 1980's, oligodeoxynucleotides (ODNs) with unique chemistries were tested with model systems both in vitro and in vivo with varying degrees of success. In the 1990's, ribozymes with both antisense and catalytic properties were successfully introduced to the field. Ribozymes were shown to selectively knock down targeted genes in human tumors grown in mice but delivery issues for these therapeutic anti-genes limited their clinical utility. Short interfering RNA (siRNA) is currently the fastest growing sector of this anti-gene field for target validation and therapeutic applications. The siRNA field may have an opportunity to impact the clinic faster than antisense and ribozymes if the scientists can overcome the previous anti-gene limitations. Fortuitously, there have been a several developments involving the expansion of our genomic knowledge coupled with the rapid dissemination of disease genes by the digital revolution. This convergence of the knowledge of the human genome with the speed of digital communication will help facilitate swift changes in the detection and treatment of human illnesses. The anti-gene field is positioned to exploit this timely union of two distinct technologies. Anti-gene molecules have an opportunity to become a successful technology in understanding the human genome, as well as, enabling the development of efficacious gene therapy for human diseases in the near future. This review will characterize the advances in this field and address the challenges to the success of for the anti-gene technology. PMID- 15544490 TI - Oligonucleotide mediated gene targeting in mammalian cells. AB - Gene targeting can be loosely defined as a process through which a specific chromosomal sequence is recognized and bound by a reagent designed for the purpose. The endpoint may be modulation of events at the target, such as transcription, or a permanent change in sequence at the site. A facile strategy for mammalian cells would have broad applications in basic and applied research, including gene therapy. Although approaches based on homologous recombination are routinely employed for transgenic animal construction, they are too laborious and inefficient for broader use. Consequently there has been a longstanding interest in developing effective synthetic reagents. Sequence recognition can be either at the level of a single strand or via the major or minor grooves, and specific approaches for each route are under development. In this review several oligonucleotide-based strategies will be discussed. These include single and double strand oligonucleotides designed to attack single strand targets, and triple helix forming olgonucleotides and peptide nucleic acids, intended for double stranded targets. PMID- 15544491 TI - Neutrally charged phosphorodiamidate morpholino antisense oligomers: uptake, efficacy and pharmacokinetics. AB - Antisense technology constitutes development of sequence-specific DNA or RNA analogs that can block the activity of selected single-stranded genetic sequences and offer the potential of high specificity lacking in many current drug treatments. The sequencing of the human genome has greatly increased the potential of this approach. Antisense oligonucleotides, the most commonly used antisense approach, are unmodified or chemically modified single stranded RNA or DNA molecules specifically designed to hybridize to corresponding RNA by Watson Crick binding. Phosphorodiamidate Morpholino oligomers (PMO) are a novel class of non-ionic antisense agents that inhibit gene expression by binding to RNA and sterically blocking processing or translation. PMOs have shown excellent efficiency and safety profile via various routes of administration in multiple animal and human studies. This review will summarize the preclinical studies with PMOs on the road to their development as therapeutic agents with particular emphasis on in vivo biodistribution and pharmacokinetics. PMID- 15544492 TI - The ins and outs of RNAi in mammalian cells. AB - The ability to utilize the RNA interference (RNAi) machinery for silencing target gene expression has created a lot of excitement in the research community. RNAi in mammalian cells is achieved through introduction or expression of 21-23 bp small interfering RNAs (siRNAs) in cells or animals. Currently, there are six ways of producing siRNAs. siRNAs can be produced by chemical synthesis, in vitro transcription, or RNase III/Dicer digestion of long dsRNAs. Alternatively, they can be expressed in vivo from plasmids, PCR cassettes, or viral vectors that include a CMV or polymerase III (pol III) transcription unit. So far, these approaches have been used to create siRNAs for use in loss-of-function studies. However, it is clear that siRNAs also hold great promise as therapeutic tools. First, their activity seems to be very sequence-specific. Moreover, siRNAs could be modified in order to increase their stability and potency in vivo. Here, we will review the issues and findings related to siRNA design and production. Moreover, we will summarize new findings on siRNA specificity, modification, and delivery, which are critical to their use as therapeutic agents. PMID- 15544493 TI - SV40 pseudovirions as highly efficient vectors for gene transfer and their potential application in cancer therapy. AB - Among viral and non-viral gene delivery systems, SV40-based vectors show great promise in the cancer gene therapy field. SV40 vectors very efficiently deliver genes such as anti-viral agents, DNA vaccine, genes for chemoprotection (such as ABC transporters genes), suicide genes and antiangiogenic genes. The recombinant SV40 vectors can infect a wide variety of cells-dividing cells as well as non cycling ones. Most of the SV40-based vectors can incorporate larger transgenes than the capacity of the SV40 wild-type, which is 5.2 kb; Moreover, in vitro packaged vectors demonstrate efficient delivery of plasmids with a molecular weight of up to 17.7 kb. SV40-based vectors carry some SV40 viral sequences, but the SV40 in vitro-packaged vectors are free of any SV40 wild-type viral DNA sequences. These vectors are prepared with nuclear extracts of SF9 insect cells containing the main viral capsid protein of the SV40 wild-type virus, VP1. This review summarizes different strategies in which SV40 vectors are used to deliver genes in vitro, to living mice, and to tumors growing in nude mice. PMID- 15544494 TI - Targeting steroid hormone receptor pathways in the treatment of hormone dependent cancers. AB - Sex steroid hormones play a central role in the development and progression of prostate and breast cancers. The biological functions of these and other steroid hormones are mediated by a family of closely related steroid hormone receptors (SHRs), with the androgen receptor (AR) mediating the effects of testosterone and related androgens, and the classical estrogen receptor (ERalpha) mediating the effects of estradiol. Recent studies have begun to elucidate the complex pathways through which SHRs regulate gene expression, and their interaction with other cellular pathways. These studies have also begun to reveal molecular mechanisms underlying the diverse spectrum of effects mediated by steroid hormone analogues in different tissues. A major advance has been the finding that certain drugs induce unique conformational changes in SHRs that alter their interactions with transcriptional coactivator and corepressor proteins, resulting in cell type specific responses. These unique conformational changes appear responsible for the tissue specific effects of the selective estrogen receptor modulators (SERMs) in breast cancer. SHRs are clearly well established therapeutic targets in cancer, and drug development has continued to focus on agents that either block steroid hormone production or bind to and modulate their receptors. The identification of multiple proteins and pathways that mediate the downstream functions of SHRs may eventually provide additional therapeutic targets. This review outlines the basic biology of SHR structure and function, with a focus on AR and ERalpha. Hormonal therapies in prostate and breast cancer that directly target AR and ERalpha, respectively, are then presented and possible novel drug targets in the SHR pathway are discussed. PMID- 15544495 TI - Role of genomics-based strategies in overcoming chemotherapeutic resistance. AB - As cancer is being recognized as a failure of apoptosis, apoptosis-based strategies are being designed. Caspases are critical for the induction of apoptosis and their decreased expression is correlated with increased grade of cancer, while increased expression of caspases rendered the cancer cells susceptible to chemotherapy. However, the endogenous functions of caspases are inhibited by inhibitors of apoptosis (IAPs) that bind activated caspases. Methods to suppress the function of IAP induced apoptosis in chemo-resistant cancer cells. The function of IAPs is inhibited by Second Mitochondria-Derived Activator Of Caspase (Smac) or Direct IAP Binding Protein With Low Pi (DIABLO). Upon apoptotic stimulus Smac/DIABLO is released from the mitochondria, which binds to IAPs and inhibits their caspase-binding activity. Overexpression of Smac/DIABLO sensitized neuroblastoma to TRAIL (TNFalpha-Related Apoptosis-Inducing Ligand). Activation of TRAIL pathway has become an important method of inducing apoptosis except in TRAIL-resistant cells. However, treatment of these cells with other cytotoxic drugs sensitizes them to TRAIL, providing effective therapeutic advantages. In addition to activating apoptotic pathways, inhibiting or suppression of cell proliferation is necessary to sensitize cancer cells to apoptosis. Critical among these proteins are NFkappaB and Akt. NFkappaB blocked apoptosis by interfering with the function of TNFalpha/TRAIL and/or through the activation of antiapoptotic proteins of the Bcl2 family. Similarly, Akt mediate cell survival via the regulation of cell survival proteins and by blocking the function of proapoptotic Bad by phosphorylation. Altering the expression of Akt by dominant negative constructs or by expression of PTEN interferes with Akt function. In summary, this review points out the complexity of interactions of the cell survival and death pathways and highlights some methods to manipulate them to achieve therapeutic advantage. PMID- 15544496 TI - The macrolide antibiotics: a pharmacokinetic and pharmacodynamic overview. AB - The macrolide antimicrobial family is comprised of 14, 15 and 16 member-ringed compounds that are characterized by similar chemical structures, mechanisms of action and resistance, but vary in the different pharmacokinetic parameters, and spectrum of activity. The macrolides accumulate in many tissues such as the epithelial lining fluid and easily enter the host defense cells, predominantly macrophages and polymorphonuclear leukocytes (PMNs). Concentrations of the macrolides in respiratory tract tissues and extracellular fluids are in almost all cases higher than simultaneously measured serum concentrations, making them useful for respiratory tract infections. This review will focus on pharmacokinetic and pharmacodynamic aspects of the clinical relevant macrolides including azithromycin, clarithromycin, dirithromycin, erythromycin and roxithromycin. PMID- 15544497 TI - Regulatory effects of macrolides on bacterial virulence: potential role as quorum sensing inhibitors. AB - Pseudomonas aeruginosa is an opportunistic pathogen, and this organism is a major cause of pulmonary damage and mortality in patients with cystic fibrosis (CF), diffuse panbronchiolitis (DPB) and other forms of bronchiectasis. A break-through in the treatment of DPB and associated chronic P. aeruginosa pulmonary infection was realized when a patient with DPB improved dramatically after treatment with erythromycin for years. Now, long-term macrolide therapy has become a first line of treatment in DPB patients, and the immunomodulatory properties have now been extended to other clinical settings, including CF. An important factor in the pathogenesis of chronic P. aeruginosa infection is a bacterial cell-to-cell signaling mechanism, referred to as "quorum sensing", which enables bacteria to coordinately turn on and off specific virulence genes through the production of autoinducer molecules. Interference or blocking of quorum-sensing systems has been considered an attractive therapeutic strategy. Clinical and basic science data suggests the potential of macrolides as relevant inhibitors of the Pseudomonas quorum-sensing system. In fact, certain macrolides strongly suppressed quorum-sensing associated genes and autoinducer production, in addition to inhibition of a variety of virulence factors. In this review, clinical efficacy of macrolides on DPB and CF patients will be briefly summarized. Additionally, the mechanisms of action of macrolides will be discussed from the standpoint of sub-MIC macrolide effects on P. aeruginosa, particularly the ability of this antibiotic to suppress quorum-sensing systems, which may be crucial in the pathogenesis of chronic P. aeruginosa infection. PMID- 15544498 TI - Cellular and molecular effects of macrolides on leukocyte function. AB - Macrolide antimicrobials have stimulated worldwide interest owing to their therapeutic effects in various inflammatory, apparently non infectious, diseases. Abundant data are now available on their interactions with host cell (specially phagocyte) functions. Modulation of oxidant production by neutrophils and of pro inflammatory cytokine synthesis and release by leukocytes are the two main effects observed in vitro. However, despite an extensive literature, many questions remain, such as the cellular/microbial target(s) of macrolide action, the critical chemical structure(s), and the usefulness of combining antimicrobial and anti-inflammatory properties, which during long-term treatment, could lead to increased microbial resistance. Also, because of the multiplicity of macrolide effects on different cell subsets, a unifying hypothesis for macrolide interactions with host cells is lacking. Novel analytical methods will certainly lead to new macrolide-based therapeutic strategies in cancer and inflammation. PMID- 15544499 TI - Immunomodulatory effects of macrolides in the lung: lessons from in-vitro and in vivo models. AB - Macrolide antibiotics appear to play a role in the management of diseases of chronic airway inflammation, distinctly separate from their antibactericidal activity. In the last fifteen years, their success in human clinical trials has prompted both in-vitro and in-vivo investigations to determine the mechanisms by which this family of antibiotics modulate the immune response. A large body of evidence suggests that macrolides directly target multiple components of the inflammatory cascade that occur independent of bactericidal/bacteriostatic effects. We will review the existing data in support of immunomodulatory effects of macrolides on activated leukocytes at the site of lung inflammation, on pulmonary host cells, and in animal models of lung disease. PMID- 15544500 TI - Clinical implications of macrolide therapy in chronic sinopulmonary diseases. AB - Macrolides have broad antibacterial spectrum and proven efficacy in the management of respiratory tract infections. Over the past decade there has been progressive interest in these agents for their potential role as tissue modifying, anti-inflammatory agents. Increasingly, the effect of macrolides on numerous cell types has been documented. Preliminary data have suggested a beneficial clinical role of chronic macrolide therapy in selected patients with chronic rhinosinusitis, chronic obstructive pulmonary disease, cystic fibrosis, non-cystic fibrosis bronchiectasis and asthma. This review presents the biological rationale and the available clinical data of chronic macrolide therapy in chronic respiratory tract diseases. When available, the data addressing the presumed mechanisms underlying clinical benefits are presented. PMID- 15544501 TI - Rationale for the development of cholinesterase inhibitors as anti-Alzheimer agents. AB - Alzheimer's disease (AD) is characterized by progressive dementia caused by the loss of the presynaptic markers of the cholinergic system in the brain areas related to memory and learning and brain deposits of amyloid beta peptide (A beta) and neurofibrillary tangles (NFT). A small fraction of early onset familial AD (FAD) is caused by mutations in genes, such as the beta-amyloid precursor protein (APP) and presenilins that increase the load of A beta in the brain. These studies together with findings that A beta is neurotoxic in vitro, provide evidence that some aggregates of this peptide are the key to the pathogenesis of AD. The yield of A beta and the processing and turnover of APP are regulated by a number of pathways including apolipoprotein E, cholesterol and cholinergic agonists. Early studies showed that muscarinic agonists increased APP processing within the A beta sequence (sAPP alpha). More recently, we have presented evidence showing that some, but not all, anticholinesterases reduce secretion of sAPP alpha as well as A beta into the media suggesting that cholinergic agonists modulate A beta levels by multiple mechanisms. Herein we review the recent advances in understanding the function of cholinesterase (ChE) in the brain and the use of ChE-inhibitors in AD. We propose and support the position that the influence of cholinergic stimulation on amyloid formation is critical in light of the early targeting of the cholinergic basal forebrain in AD and the possibility that maintenance of this cholinergic tone might slow amyloid deposition. In this context, the dual action of certain cholinesterase inhibitors on their ability to increase acetylcholine levels and decrease amyloid burden assumes significance as it may identify a single drug to both arrest the progression of the disease as well as treat its symptoms. A new generation of acetyl- and butyryl cholinesterase inhibitors is being studied and tested in human clinical trials for AD. We critically discuss recent trends in AD research, from molecular and genetic to clinical areas, as it relates to the effects of cholinergic agents and their secondary effects on A beta. Finally, we examine different neurobiological mechanisms that provide the basis of new targets for AD drug development. PMID- 15544502 TI - An overview of the current and novel drugs for Alzheimer's disease with particular reference to anti-cholinesterase compounds. AB - Several cellular processes could be targeted if the complex nature of Alzheimer's disease (AD) was already understood. Most of AD treatments have been focused on the inhibition of acetylcholinesterase (AChE) in order to raise the levels of its substrate, i.e. the neurotransmitter acetylcholine (ACh), to augment cognitive functions of affected patients. Effectiveness in AChE inhibition and side-effect issues of clinical (tacrine, donepezil, galanthamine and rivastigmine) as well as of novel inhibitors is reviewed here. Novel design methods for the inhibition of AChE include the use of in silico tools to predict the interactions between AChE and the desired compound, both at the active site of the enzyme, responsible of hydrolysing ACh and with the peripheral anionic site (PAS), which has been described as a promoting agent of the amyloid beta-peptide (A beta) aggregation present in the senile plaques of the brain of AD individuals. PMID- 15544503 TI - Molecular modelling approaches to the design of acetylcholinesterase inhibitors: new challenges for the treatment of Alzheimer's disease. AB - The interest for acetylcholinesterase as a target for the palliative treatment of Alzheimer's disease has been renewed in the last years owing to the evidences that support the role of this enzyme in accelerating the aggregation and deposition of the beta-amyloid peptide. A large amount of structural information on the acetylcholinesterase enzyme and of its complexes with inhibitors acting at the catalytic site, the peripheral binding site, or both is now available. Based on that, molecular modelling studies can be intensively used to decipher the molecular determinants that mediate the relationship between chemical structure and inhibitory potency. In turn, this knowledge can be exploited to design new compounds leading to more effective cholinergic strategies. At this point, inhibitors able to interact at the peripheral binding site are of particular relevance, as they might disrupt the interactions between the enzyme acetylcholinesterase and the beta-amyloid peptide. Therefore, these compounds might not only ameliorate the cholinergic deficit, but also be capable of slowing down the progression of the disease. PMID- 15544504 TI - Development of bivalent acetylcholinesterase inhibitors as potential therapeutic drugs for Alzheimer's disease. AB - At present the only FDA-approved therapy for Alzheimer's disease involves the administration of acetylcholinesterase inhibitors, to alleviate the cholinergic deficit associated with this disease. However, none of the approved drugs is ideal in efficacy or tolerability. One possible strategy to improve selectivity and potency is to design drugs that can simultaneously bind to the catalytic and peripheral anionic sites of AChE. In this review we will describe the development of dimeric AChE inhibitors, from the early observations of high inhibition potency by bis-quaternary inhibitors, to the structure-based design of dimers based on tacrine, huperzine A, galanthamine, and polyamines. PMID- 15544505 TI - Acetylcholinesterase inhibitors as a starting point towards improved Alzheimer's disease therapeutics. AB - The knowledge about the pathogenesis and the development of the neurodegeneration associated with Alzheimer's disease (AD) has been organised throughout the years into two theories, namely the cholinergic and the amyloid hypotheses. The loss of cholinergic neurotransmission and the abnormal aggregation and deposition of the amyloid-beta peptide (A beta) in the brain are retained as the central events by the two theories, respectively. These phenomena and their pathological consequences are the main targets of the drug discovery strategies based on each hypothesis. However, the two paradigms share some common aspects as shown by several experimental evidences, such that they might even fit into a unifying scenario of neuropathology and neurodegeneration. In this context, in a perspective of drug discovery, the enzyme acetylcholinesterase (AChE) holds a key position, as it is a main target for cholinomimetic AD drugs being responsible for the breakdown of the neurotransmitter, and it is also involved in the aggregation of A beta and the formation of the neurotoxic fibrils. Following this view, in recent years, a drug design strategy has emerged, directed to finding molecules able to inhibit both of these actions exerted by AChE. In this review, we will briefly introduce the biological basis of this strategy, and then will account for the early results obtained in this field in our and in other laboratories. The main focus will be on potential lead compounds for which some experimental evidence exists supporting the hypothesis of their dual action, as AChE inhibitors and blockers of the AChE-induced A beta aggregation. PMID- 15544506 TI - Recent approaches to novel anti-Alzheimer therapy. AB - Insufficient cholinergic neurotransmission in AD is responsible for a progressive loss of cognition and motor capacities. The cholinergic hypothesis has provided the rationale for the current treatment approaches based on acetylcholinesterase inhibitors. However, recent data focus on the complex nature of AD and disclose the involvement of other neurotransmitters such as serotonin, noradrenalin, dopamine, histamine, excitatory amino acids and neuropeptides among others. Interestingly, recent research has revealed that in severe AD brains the levels of AChE are considerably reduced whereas BuChE activity increases, thus aggravating the toxicity of beta A. In such instances, it is possible that BuChE may be a more suitable target than AChE. Oxidative stress has been implicated in CNS degenerative disorders such as AD and PD. Therefore, owing to their capacity to inhibit oxidative damage, MAOIs are potential candidates as anti-Alzheimer drugs. More recently, a novel drug--TV3326--was designed, based upon two pharmacophores: the carbamate moiety from rivastigmine, an AChE inhibitor; and the propargyl group from rasagiline, a MAO inhibitor. This drug exhibits cholinesterase and selective brain MAO inhibitory activities, reduces apoptosis and stimulates the processing of APP alpha, hence reducing the possibility of generation of the toxic beta A. Thus, TV3326 may be expected to contribute positively to the cognitive benefits of Alzheimer's patients. Anyhow, the development of drugs with several targets and diverse pharmacological properties may conclusively demonstrate the most beneficial therapy. PMID- 15544507 TI - New classes of AChE inhibitors with additional pharmacological effects of interest for the treatment of Alzheimer's disease. AB - Alzheimer's disease (AD) is associated to a gradual loss of attention and memory that have been associated to impairment of brain cholinergic neurotransmission, particularly a deficit of cholinergic neurons in the nucleus basalis of Meynert. Thus, it is not surprising that the first therapeutic target that has demonstrated therapeutic efficacy on cognition, behaviour and functional daily activities has been the inhibitors of acetylcholinesterase (AChE), i.e. tacrine, donepezil, rivastigmine and galanthamine. But not all inhibitors of AChE have the same potency to block the enzyme and have a different pharmacological profile. For instance, rivastigmine is a dual inhibitor of AChE and butyrylcholinesterase (BuChE), and galanthamine is a mild inhibitor of AChE and an allosteric potentiating ligand of neuronal nicotinic receptors for acetylcholine (nAChRs). In addition, we have recently found that galanthamine has neuroprotective effects by inducing calcium signals and the induction of the antiapoptotic protein Bcl-2. In this frame, we have been synthesizing new tacrine derivatives that keep their ability to inhibit AChE but that interfere with neuronal calcium overloading and prevent apoptosis. Some of these compounds exhibit neuroprotecting properties and thus, could be useful in the treatment of neurodegenerative and ischaemic brain diseases. PMID- 15544508 TI - Identification and validation of novel drug targets in tuberculosis. AB - There is an urgent need for new antimycobacterial drugs, and in particular for novel agents that will shorten the duration of tuberculosis chemotherapy, or overcome drug-resistant strains of the causative organism, Mycobacterium tuberculosis. Our knowledge of the tubercle bacillus and its complex interaction with the human host has improved dramatically in recent years, particularly with the determination of its complete genome sequence. New genome-scale tools are being applied to aid in drug target identification, alongside traditional approaches aimed at understanding the basic biology of M. tuberculosis. Many potential drug targets have been identified, but very few have been validated by showing that they are essential for growth or survival of the bacterium. In this review, the landscape of potential drug targets is surveyed.Structural Bioinformatic Approaches to the Discovery of New Antimycobacterial Drugs. PMID- 15544509 TI - Structural bioinformatic approaches to the discovery of new antimycobacterial drugs. AB - Integrated bioinformatic approaches to drug discovery exploit computational techniques to examine the flow of information from genome to structure to function. Informatics is being be used to accelerate and rationalize the process of antimycobacterial drug discovery and design, with the immediate goals to identify viable drug targets and produce a set of critically evaluated protein target models and corresponding set of probable lead compounds. Bioinformatic approaches are being successfully applied in the selection and prioritization of putative mycobacterial drug target genes; computational modelling and x-ray structure validation of protein targets with drug lead compounds; simulated docking and virtual screening of potential lead compounds; and lead validation and optimization using structure-activity and structure-function relationships. By identifying active sites, characterizing patterns of conserved residues and, where relevant, predicting catalytic residues, bioinformatics provides information to aid the design of selective and efficacious pharmacophores. In this review, we describe selected recent progress in antimycobacterial drug design, illustrating the strengths and limitations of current structural bioinformatic approaches as tools in the fight against tuberculosis. PMID- 15544510 TI - Fluoroquinolones as chemotherapeutics against mycobacterial infections. AB - The antimicrobial agents used in the treatment of mycobacterial infections have remained largely unchanged for several decades. Primary treatment of tuberculosis relies on four drugs, isoniazid, a rifamycin, pyrazinamide, and ethambutol (or streptomycin), and generally results in >95% cure in uncomplicated tuberculosis infection. Drug resistance greatly complicates treatment of this disease. Treatment of tuberculosis caused by multiply drug-resistant strains with "second line" drugs remains complex, and is generally tailored to the individual patient and strain. Several of the fluoroquinolones have shown promise as second line drugs for treatment of active disease and, in combination with clarithromycin or azithromycin, ethambutol, and other agents, for treatment of Mycobacterium avium complex infection. While large clinical trials are not possible with second line drugs, clinical treatment data are available and suggest that the quinolones have various degrees of promise in treatment of these infections. Bacterial type II DNA topoisomerases, DNA gyrase and topoisomerase IV, are the targets of quinolones, and provide the genetic basis for quinolone activity in mycobacteria. Mutations in these enzymes results in resistance, and characterization of resistant mutants allows correlation of genotype with susceptibility phenotype. Structure-activity relationship studies have provided further insight into optimal use of quinolones in mycobacterial infections. Care should be taken in treating pneumonia with fluoroquinolones if there is a degree of suspicion of tuberculosis, since quinolone monotherapy may rapidly select for quinolone resistance, thereby removing that class of antibiotic from the small range of treatment options. PMID- 15544511 TI - Effects of macrolides and ketolides on mycobacterial infections. AB - New macrolides, such as clarithromycin and azithromycin, are active agents to Mycobacterium avium complex (MAC). Both clarithromycin and azithromycin are well known for the ability to improve the prognosis of AIDS patients with disseminated MAC infection. However, the administration of monotherapy with a macrolide is usually associated with the emergence of drug resistance after a few months of use. Therefore, the recommended treatment for MAC infection involved the use of at least two antibiotics, which includes a macrolide in combination with rifabutin, moxifloxacin and/or ethambutol. When used as prophylactic therapy in AIDS patients, azithromycin is more convenient (1200 mg, once a week) than clarithromycin (500 mg, twice a day). Ketolides are a semi-synthetic derivative of erythromycin A, which differs from erythromycin A by substitution of a 3-keto group for L-cladinose. Telithromycin has a carbamate group linked to an imidazolium and pyridium nucleus at C11-C12. In mice model, both telithromycin and ABT-733 were active in vivo against MAC. PMID- 15544512 TI - In vitro and in vivo activities of new rifamycin derivatives against mycobacterial infections. AB - Several rifamycin derivatives have been developed during the last 15 years for the treatment of mycobacterial infections. For tuberculosis, rifabutin (RFB) showed strong activity and seemed to be suitable when tuberculosis patients were also treated for their AIDS infection. Rifapentine (RPT) was evaluated in patients with or without AIDS for its intermittent use. It displayed promising activity but must be strengthened in situations, such as AIDS or patients without AIDS but with cavities. Rifalazil (RLZ) has been evaluated in mice but the dosages used were much higher than those tolerated by patients. Regarding Mycobacterium avium infections, RFB showed significant prophylactic activity in humans, RPT displayed some activity in mice and RLZ showed modest activity in mice. PMID- 15544513 TI - Prospects for clinical introduction of nitroimidazole antibiotics for the treatment of tuberculosis. AB - Nitroaromatic antibiotics have a long and controversial history in human and veterinary medicine. This controversy lies behind the presumption of many pharmaceutical companies that nitroaromatic compounds should be filtered from the list of drug-like compounds but stands at odds with the remarkably safe clinical record of use of such compounds. In this review, we will describe the whole-cell structure-activity relationships that have been reported for antimycobacterial nitroimidazoles as well as the available in vivo data supporting efficacy with a particular emphasis on nitroimidazo[2,1-b]oxazines such as PA-824. We will also explore the unique potential of such compounds to shorten the course of tuberculosis therapy by exerting a bactericidal effect on non-replicating bacilli. We will consider the mode of action of such compounds in sensitive organisms and discuss the mechanisms by which resistance may emerge. Finally, we will review the pharmacokinetics, toxicology and laboratory and animal studies linking nitroimidazoles with carcinogenicity and mutagenicity and assess the prospects for the clinical introduction of nitroimidazoles for the treatment of tuberculosis. PMID- 15544514 TI - Liposome technology for drug delivery against mycobacterial infections. AB - Mycobacteria are intracellular pathogens that invade and reside inside macrophages. There has been a rapid resurgence in infections caused by the genus mycobacteria. Chemotherapy of mycobacterial infections is prolonged, hepatotoxic and very often inadequate in achieving optimal drug concentrations inside the cells. Recent advances in controlled delivery systems for drugs such as liposomes have sparked a renewed interest in their potential application for the treatment of mycobacterial infections. The versatility of liposomes in incorporation of hydrophilic/hydrophobic components, non-toxic nature, biodegradability, biocompatibility and property of sustained release makes them attractive candidates for the delivery of antitubercular drugs. Liposome research in the area of mycobacterial diseases has evolved and matured through several phases; from the laboratory to the clinics. This review, thus focuses on the use of liposomes for the treatment of various types of mycobacterial diseases. PMID- 15544515 TI - Microsphere technology for chemotherapy of mycobacterial infections. AB - The purpose of this review article is to examine the various studies that have evaluated microspheres for delivery of antimycobacterial drugs. Some of the studies strictly involve the development and evaluation of microspheres for use in antimycobacterial drug delivery, whereas others actually use drug-loaded microspheres to treat mycobacterial infections in cell lines and small animals. Although there is a potential to use microspheres to treat a variety of mycobacterial infections, it appears that most of the studies so far have focused on the etiological agent of tuberculosis, Mycobacterium tuberculosis. As a result, the infectious studies presented here all entail the treatment of that mycobacterial agent. This review will address the following aspects that are important if microspheres are to be considered an acceptable therapeutic tool: 1) in vitro release characteristics, 2) delivery, release and efficacy in macrophages, 3) effectiveness in infected small animal models, 4) safe and combined use with other antimycobacterial agents, and 5) reduced toxicity. It is hoped that once all of these parameters are evaluated, a conclusion regarding the benefit of microsphere technology in the treatment of mycobacterial diseases can be reached. PMID- 15544516 TI - Predictive in vitro models of the sterilizing activity of anti-tuberculosis drugs. AB - Sterilizing drugs kill Mycobacterium tuberculosis that persists during chemotherapy. Predictive models should mimic the conditions causing persistence in the lesions of cavitary disease, and should grade current anti-tuberculosis drugs according to their sterilizing activity determined in clinical trials. Models should start with old, stationary cultures grown micro-aerophilically. In these, persistent bacilli occur in different populations in which there is no appreciable cell division. Population 1. Grows in liquid culture medium but not on solid medium. Killed by rifampicin. Population 2. Grows on solid culture medium. Killed by rifampicin. Population 3. Grows in liquid medium but not on solid medium. Tolerant of rifampicin. Population 4. Bacilli from Cornell model mice, after treatment with pyrazinamide and isoniazid, cannot grow in liquid or on solid culture medium. Some of these populations are incorporated in models which start with 100-day liquid medium cultures. In model 1 (population 2) the new drug is added and colony counted after 7 days incubation. In models 2 and 3, 100 mg/L rifampicin is added to the 100-day culture when the bacilli lose their ability to grow on solid culture medium (population 3). After re-suspension in rifampicin-free liquid medium for 7 days, the bacilli recover growth on solid medium, when a colony count is done. The new drug is added during incubation with rifampicin in model 3 and at the start of recovery in drug-free medium in model 2. Models 1 and 3 grade isoniazid, rifampicin and pyrazinamide according to their sterilizing activity determined by clinical trials. PMID- 15544517 TI - Adjunctive immunotherapy of mycobacterial infections. AB - In order to cope with the worldwide increase in the prevalence of multidrug resistant tuberculosis and Mycobacterium avium complex (MAC) infections, a number of new antimycobacterial drugs have been or are being synthesized and developed. Development of new protocols for chemotherapy of refractory mycobacterioses is also sharing promise. In this context, one promising strategy is to devise regimens to treat patients with refractory mycobacterioses using ordinary antimycobacterial agents in combination with appropriate immunomodulators. This article deals with the following matters: an outline of the host immune response to mycobacterial pathogens, particularly in terms of mobilization of the cytokine network in response to mycobacterial infection, and adjunctive immunotherapy using (1) recombinant immunomodulating cytokines, (especially Th-1 and Th-1-like cytokines such as IFN-gamma, IL-2, IL-12, IL-18 and GM-CSF), (2) inhibitors of immunosuppressive cytokines (TGF-beta) and some proinflammatory tissue-damaging cytokines (TNF-alpha), and (3) immunomodulatory agents such as ATP and its analogs, imidazoquinoline, diethyldithiocarbamate, poloxamer, dibenzopyran, galactosylceramide, nonsteroidal anti-inflammatory drugs, Chinese traditional medicines, levamisole, synthesized mycobacterial oligoDNA, DNA vaccine expressing mycobacterial HSP65 or IL-12, and heat-killed Mycobacterium vaccae. Although adjunctive immunotherapy is fairly efficacious in treating intractable mycobacterioses, it still features serious problems and dilemmas, such as high cost, occasionally severe side effects, and, in many cases, only modest efficacy in potentiating host defense mechanisms against mycobacterial infections, primarily because of the induction of macrophage-deactivating cytokines during the course of long-term administration of adjunctive agents. PMID- 15544518 TI - Angiotensin and bradykinin: targets for the treatment of vascular and neuro-glial pathology in diabetic retinopathy. AB - The renin-angiotensin system (RAS) and kallikrein-kinin system (KKS) are complex pathways linked by a number of molecules that participate in both systems. Apart from modulating a variety of normal physiological processes, both the RAS and KKS are up-regulated following tissue injury where they influence vascular function, inflammation, cell growth and differentiation and angiogenesis. The RAS exerts its effects by the generation of a family of bioactive angiotensin peptides in which angiotensin II (ANG II) and the angiotensin type 1 (AT1) and angiotensin type 2 (AT2) receptors are most well characterised. In the KKS, bradykinin (BK) and kallidin and their carboxypeptidase metabolites, des-Arg(9)-BK and des Arg(10)-kallidin, are the effector peptides exerting their actions via BK type 1 (BK-B1) and BK type 2 (BK-B2) receptors. Emerging evidence suggests that an ocular RAS is activated in diabetic retinopathy and may contribute to progressive alterations to retinal cells such as pericytes, endothelial cells, neurons and glia. Less well studied is the retinal KKS, however recent studies indicate effects on retinal electrophysiology and angiogenesis. The pathogenetic actions of the RAS and KKS in many tissues and possibly the diabetic retina are mediated by specific growth factors such as vascular endothelial growth factor (VEGF) and connective tissue growth factor (CTGF). This review will examine the roles of the RAS and KKS in both retinal vascular and neuro-glial dysfunction in diabetic retinopathy, and the potential of blockade of these systems for the prevention and treatment of this serious diabetic complication. PMID- 15544519 TI - Vasoactive factors and diabetic retinopathy: vascular endothelial growth factor, cycoloxygenase-2 and nitric oxide. AB - The pathogenetic agents involved in the development of diabetic retinopathy are diverse. In the setting of hyperglycaemia and retinal hypoxia a number of vasoactive factors may interact to promote pathology in a variety of cell types including the microvasculature, neurons and glia. Vascular endothelial growth factor (VEGF) is universally accepted as a primary factor in the regulation of vessel patency in vascular networks throughout the body and including the retina. There is considerable evidence that the VEGF system in disturbed early in diabetes and interacts with other pathways and vasoactive factors to stimulate breakdown of the blood retinal barrier (BRB) and eventually promote angiogenesis, the hallmark feature of proliferative diabetic retinopathy (PDR). There is a distinct relationship between VEGF and the prostaglandin-cyclooxygenase system. Prostaglandins influence retinal blood flow, are important in inflammation and are also pro-angiogenic. Recent evidence suggests that cyclooxygenase-2 (COX-2) modulates angiogenesis by interacting with the VEGF system. Like prostaglandins, nitric oxide (NO) is a vasodilator and is implicated in VEGF mediated vascular permeability and angiogenesis. Emerging evidence indicates that COX-2 also interacts with NO and that these two systems have reciprocal effects on each other. There is little doubt that the interactions between these three vasoactive systems are complex and requires further study in the context of retinal vascular permeability, angiogenesis and neurodegeneration. This review will explore experimental and clinical evidence that VEGF, COX-2 and NO promote retinal pathology in diabetes and other ischemic-induced retinopathies. PMID- 15544520 TI - Advanced glycation and advanced lipoxidation: possible role in initiation and progression of diabetic retinopathy. AB - Diabetic retinopathy remains the most common microvascular complication suffered by diabetic patients and is the leading cause of registerable blindness in the working population of developed countries. The clinicopathological lesions of diabetic retinopathy have been well characterised and although a multitude of pathogenic mechanisms have been proposed, the underlying dysfunctional biochemical and molecular pathways that lead to initiation and progression of this complication remain largely unresolved. There is little doubt that the pathogenesis of diabetic retinopathy is highly complex and there is a pressing need to establish new therapeutic regimens that can effectively prevent or limit retinal microvascular cell dysfunction and death which is characteristic of the vasodegenerative stages of diabetic retinopathy. The formation and accumulation of advanced glycation endproducts (AGEs) and/or advanced lipoxidation endproducts (ALEs) are among several pathogenic mechanisms that may contribute to diabetic retinopathy. AGEs/ALEs can form on the amino groups of proteins, lipids and DNA through a number of complex pathways including non-enzymatic glycation by glucose and reaction with metabolic intermediates and reactive dicarbonyl intermediates. These reactions not only modify the structure and function of proteins, but also cause intra-molecular and intermolecular cross-link formation. AGEs/ALEs are known to accumulate in the diabetic retina where they may have important effects on retinal vascular cell function, as determined by a growing number of in vitro and in vivo studies. Evidence now points towards a pathogenic role for advanced glycation/lipoxidation in the initiation and progression of diabetic retinopathy and this review will examine the current state of knowledge of AGE/ALE-related pathology in the diabetic retina at a cellular and molecular level. It will also outline how recent pharmaceutical strategies to inhibit AGE/ALE formation or limit their pathogenic influence during chronic hyperglycaemia may play a significant role in the treatment of diabetic retinopathy. PMID- 15544521 TI - Advanced glycation: how are we progressing to combat this web of sugar anomalies in diabetic nephropathy. AB - Advanced glycation end products (AGEs) in diabetic nephropathy have been extensively researched over the last decade and are now firmly established as major players in this disease. The enigma remains the search for the ideal AGE inhibition therapy, which is a great challenge in the context of the structural diversity inherent to AGE chemistry. Certainly, there is a requirement to standardize measurements of circulating and tissue levels of AGEs and to characterize the pathogenic potential of specific AGE moieties. In order to develop more effective, targeted approaches to combat diabetic nephropathy, the mechanisms of action of selective AGE inhibitors and the inter-relationships of advanced glycation with other pathogenic pathways must be addressed. PMID- 15544522 TI - Vasoactive renal factors and the progression of diabetic nephropathy. AB - Ongoing investigation into the relationship between the renin-angiotensin system (RAS) and the progression of diabetic renal disease has persisted for the past two decades. Experimental and clinical evidence suggests that the RAS has a pathogenic role, induced by its haemodynamic and non-haemodynamic mechanisms. The discovery of a local intrarenal RAS provides a rationale for investigating the components of RAS, specifically Angiotensin II (AngII) in the diabetic setting. AngII has multiple effects, including activating intracellular second messengers, transcription factors, extracellular matrix protein and also growth factors and cytokines, which lead to many of the structural and functional changes in the diabetic kidney. The beneficial effects afforded by RAS blockade further implicate AngII in the progression of diabetic nephropathy. Although AngII is a common suspect in the pathogenesis of diabetic nephropathy RAS blockade does not prevent patients from progressing to end stage renal disease. Evaluating other vasoactive factors, which have similar and distinct functions to AngII, will assist in understanding their potential role in the pathogenesis of diabetic nephropathy. A large number of researchers are studying vasoactive factors, however, the case for their role in diabetic nephropathy is inconclusive. Further investigation into the effects of inhibiting vasoactive compounds, including endothelin, urotensin II and vasopeptidases, together with inhibiting RAS, may provide another therapeutic avenue for treating diabetic nephropathy. PMID- 15544523 TI - The involvement of growth hormone (GH), insulin-like growth factors (IGFs) and vascular endothelial growth factor (VEGF) in diabetic kidney disease. AB - At present, diabetic kidney disease affects about 15-25 % of patients with type 1 diabetes (T1D) and 30-40 % of patients with type 2 diabetes (T2D). Several decades of extensive research have elucidated various pathways to be implicated in the development of diabetic kidney disease. These include metabolic factors beyond blood glucose (e.g. advanced glycation endproducts (AGEs)), haemodynamic factors (e.g. the renin angiotensin system (RAS)), intracellular signaling molecule proteins (e.g. protein kinase C (PKC)) and growth factors/cytokines (e.g. growth hormone (GH), insulin-like growth factors (IGFs), transforming growth factor beta (TGF-beta) and vascular endothelial growth factor (VEGF)). This review focuses on the role of three of these growth factors, i.e. GH, IGFs and VEGF. A brief discussion of each system is followed by description of its expression in the normal kidney. Then, for each system, in vitro, experimental and clinical evidence addressing the role of the system in diabetic kidney disease is presented. The interplay of each system to other potential pathways will also be addressed. Finally, well-known and potential therapeutic strategies targeting the GH/IGF and VEGF systems in a specific or indirect way will discussed. PMID- 15544524 TI - Lipoproteins and diabetic microvascular complications. AB - Risk factors for the microvascular complications (nephropathy and retinopathy) of Type 1 and Type 2 diabetes mellitus and the associated accelerated atherosclerosis include: age, diabetes duration, genetic factors, hyperglycaemia, hypertension, smoking, inflammation, glycation and oxidative stress and dyslipoproteinaemia. Hypertriglyceridaemia, low HDL and small dense LDL are common features of Type 2 diabetes and Type 1 diabetes with poor glycaemic control or renal complications. With the expansion of knowledge and of clinical and research laboratory tools, a broader definition of 'lipid' abnormalities in diabetes is appropriate. Dyslipoproteinaemia encompasses alterations in lipid levels, lipoprotein subclass distribution, composition (including modifications such as non-enzymatic glycation and oxidative damage), lipoprotein-related enzymes, and receptor interactions and subsequent cell signaling. Alterations occur in all lipoprotein classes; chylomicrons, VLDL, LDL, HDL, and Lp(a). There is also emerging evidence implicating lipoprotein related genotypes in the development of diabetic nephropathy and retinopathy. Lipoprotein related mechanisms associated with damage to the cardiovascular system may also be relevant to damage to the renal and ocular microvasculature. Adverse tissue effects are mediated by both alterations in lipoprotein function and adverse cellular responses. Recognition and treatment of lipoprotein-related risk factors, supported by an increasing array of assays and therapeutic agents, may facilitate early recognition and treatment of high complication risk diabetic patients. Further clinical and basic research, including intervention trials, is warranted to guide clinical practice. Optimal lipoprotein management, as part of a multi-faceted approach to diabetes care, may reduce the excessive personal and economic burden of microvascular complications and the related accelerated atherosclerosis. PMID- 15544525 TI - Can an apple a day keep the doctor away? AB - The modern pharmaceutical industry based on synthetic chemistry severed the historical connection between plants, food and medicines. The growing costs of discovering new chemical entity-based drugs through high throughput screening methods may yet again reconnect plants and human health at a new level of technological sophistication. Multi-component botanical therapeutics that comprise functional foods, dietary supplements and botanical drugs hold several advantages over conventional drugs that may earn them a more prominent place in the medicine of the future. They can deliver mixtures of multi-functional molecules with potentiating and synergistic effects and pleiotropic targeting at a reasonable cost and with fewer regulatory constraints. They are well suited for long-term disease prevention in an era of genetic testing and increased life expectancy. They also provide additional vehicles for delivering health and wellness. Technologies that address the needs of discovery, development and manufacturing of multi-component botanical therapeutics are emerging. They include computational and bioinformatics approaches, cell based gene expression and high-content screening systems, and phytochemical elicitation and unique plant cultivation / extraction methods designed to optimize the production of bioactives, standardize overall extract composition and assure batch-to-batch product consistency. Nevertheless, multi-component botanical therapeutics carry risks associated with potential interactions with conventional drugs and adverse reactions, which are difficult to detect and diagnose. They face problems of acceptance by the medical community and pharmaceutical industry, safety and efficacy validation, poor standardization and quality control, and difficulties in identifying active ingredients and determining their complex mode(s) of action. Solving these problems will accelerate the merger of grocery stores with pharmacies and agriculture with chemical manufacturing and provide physicians and patients with broader and more individualized choices for disease prevention and treatment. PMID- 15544526 TI - Eph receptor tyrosine kinases in tumor and tumor microenvironment. AB - Eph receptors are a unique family of receptor tyrosine kinases (RTK) that play critical roles in embryonic patterning, neuronal targeting, and vascular development during embryogenesis. In adults, Eph RTKs and their ligands, the ephrins, are frequently overexpressed in a variety of cancers and tumor cell lines, including breast, prostate, non-small cell lung and colon cancers, melanomas, and neuroblastomas. Unlike traditional oncogenes that often function only in tumor cells, recent data show that Eph receptors mediate cell-cell interaction both in tumor cells and in tumor microenvironment, namely the tumor stroma and tumor vasculature. As such, Eph RTKs represent attractive potential targets for drug design, as targeting these molecules could attack several aspects of tumor progression simultaneously. This review will focus on recent advances in dissecting the role of Eph RTKs in tumor cells, tumor angiogenesis, and possible contribution to trafficking of inflammatory cells in cancer. PMID- 15544527 TI - Discovery, structure and biological activities of the cyclotides. AB - The cyclotides are a family of small disulfide rich proteins that have a cyclic peptide backbone and a cystine knot formed by three conserved disulfide bonds. The combination of these two structural motifs contributes to the exceptional chemical, thermal and enzymatic stability of the cyclotides, which retain bioactivity after boiling. They were initially discovered based on native medicine or screening studies associated with some of their various activities, which include uterotonic action, anti-HIV activity, neurotensin antagonism, and cytotoxicity. They are present in plants from the Rubiaceae, Violaceae and Cucurbitaceae families and their natural function in plants appears to be in host defense: they have potent activity against certain insect pests and they also have antimicrobial activity. There are currently around 50 published sequences of cyclotides and their rate of discovery has been increasing over recent years. Ultimately the family may comprise thousands of members. This article describes the background to the discovery of the cyclotides, their structural characterization, chemical synthesis, genetic origin, biological activities and potential applications in the pharmaceutical and agricultural industries. Their unique topological features make them interesting from a protein folding perspective. Because of their highly stable peptide framework they might make useful templates in drug design programs, and their insecticidal activity opens the possibility of applications in crop protection. PMID- 15544528 TI - Novel strategies for isolation and characterization of cyclotides: the discovery of bioactive macrocyclic plant polypeptides in the Violaceae. AB - This review focuses on the discovery of cyclotides in the Violaceae, their isolation and their anti-cancer effects. These macrocyclic plant peptides consist of about 30 amino acids, including three conserved disulfide bonds in a cystine knotted arrangement, which renders them a remarkable stability. Their unique structure, combined with a wide array of biological activities, makes them of great interest as possible leads in drug development or as carriers of grafted peptide sequences. Here we describe the work conducted in our laboratory, which started with the overall aim of identifying peptides and small proteins of the size 10-50 amino acid residues in plants with novel chemical structures and biological profiles with a potential for drug development or for use as pharmacological tools. Thus we developed a fractionation protocol to directly address major challenges encountered when dealing with plant material, such as removal of chlorophyll, polyphenols, and low molecular compounds omnipresent in plants. Using this protocol, we then discovered a suite of cyclotides, the varv peptides, from the plant Viola arvensis (Violaceae). Following this, separation methods directly targeting cyclotides were developed, e.g. by adsorption, ion exchange chromatography and solvent-solvent partitioning, which then were used in the isolation of additional cyclotides. To structurally examine cyclotides we have also developed methods based on mass spectrometry for cyclotide sequencing and mapping of disulfide bonds. Finally, to assess structure-activity relationships, regarding their anti-cancer and cytotoxic effects that we focus upon, we have also characterized the three dimensional structure of cyclotides by homology modeling techniques. PMID- 15544529 TI - Anti-HIV cyclotides. AB - The cyclotides are a recently discovered, structurally unique family of bioactive plant peptides. Their discovery spawned a series of structural analyses, synthetic efforts, and studies to define the biosynthesis and biological properties of these novel peptide metabolites. Cyclotides have a head-to-tail cyclized amino acid backbone and a conserved cystine knot motif that provides an extremely stable structural framework. They all share a common global fold and are highly resistant to denaturation and to cleavage by proteolytic enzymes. However, these macrocyclic peptides are quite permissive to amino acid substitutions or additions in several peripheral loop regions, since changes in these loops do not alter the core cyclotide structure. These features make cyclotides attractive templates for incorporating desired amino acid sequences and then delivering these peptide sequences in a well defined, highly stable framework. Cyclotides likely function in a defensive role in the source plants since they exhibit a broad spectrum of antimicrobial activity and are detrimental to the growth and survival of herbivorous insects. Cyclotides are gene-encoded polypeptides that are cleaved from larger precursor proteins and then cyclized. This review summarizes research done on a subset of cyclotides that were discovered due to their HIV inhibitory properties. It details the isolation and characterization of these compounds and describes this work in the context of our current state of knowledge of the entire cyclotide family. PMID- 15544530 TI - Sunflower trypsin inhibitor-1. AB - SFTI-1 is a bicyclic 14 amino acid peptide that was originally isolated from the seeds of the sunflower Helianthus annuus. It is a potent inhibitor of trypsin, with a sub-nanomolar K(i) value and is homologous to the active site region of the well-known family of serine protease inhibitors known as the Bowman-Birk trypsin inhibitors. It has a cyclic backbone that is cross-braced by a single disulfide bridge and a network of hydrogen bonds that result in a well-defined structure. SFTI-1 is amenable to chemical synthesis, allowing for the creation of synthetic variants. Alterations to the structure such as linearising the backbone or removing the disulfide bridge do not reduce the potency of SFTI-1 significantly, and minimising the peptide to as few as nine residues results in only a small decrease in reactivity. The creation of linear variants of SFTI-1 also provides a tool for investigating putative linear precursor peptides. The mechanism of biosynthesis of SFTI-1 is not yet known but it seems likely that it is a gene-coded product that has arisen from a precursor protein that may be evolutionarily related to classic Bowman-Birk inhibitors. PMID- 15544531 TI - Theta-defensins: cyclic antimicrobial peptides produced by binary ligation of truncated alpha-defensins. AB - The first cyclic peptide discovered in animals is an antimicrobial octadecapeptide that is expressed in leukocytes of rhesus monkeys. The peptide, termed rhesus Theta-defensin 1 (RTD-1) is the prototype of a new family of antimicrobial peptides, which like the previously characterized alpha- and beta defensin families, possesses broad spectrum microbicidal activities against bacteria, fungi, and protects mononuclear cells from infection by HIV-1. The cyclic Theta-defensin structure is essential for a number of its antimicrobial properties, as demonstrated by the markedly reduced microbicidal activities of de cyclized Theta-defensin analogs. Genetic and biochemical experiments disclosed that the biosynthesis of RTD-1 results from the head-to-tail joining of two nine amino acid peptides, each of which is donated by a separate precursor polypeptide, which are in fact C-terminally truncated pro-alpha-defensins. Alternate combinations of the two nonapeptides generate two additional macaque Theta-defensins, RTD-2 and RTD-3. Humans do not express Theta-defensin peptides, but mRNAs encoding at least two Theta-defensins are expressed in human bone marrow. However, in each case the open reading frame is interrupted by a stop codon in the signal peptide-coding region. The mature Theta-defensin peptide is a two-stranded beta-sheet that, like the alpha- and beta-defensins, is stabilized by three disulfides. However, the parallel orientation of the Theta-defensin disulfide arrangement allows for substantial flexibility around its short axis. Unlike alpha- and beta-defensins, RTD-1 lacks an amphiphilic topology. This may partially explain the unusual interaction between Theta-defensins and phospholipid bilayers. PMID- 15544532 TI - Retrocyclins: using past as prologue. AB - Retrocyclins are synthetic Theta-defensins that were reconstructed from genetic blueprints that had remained unused for at least 7.5 million years. From phylogenetic studies, it appears that Theta-defensins arose in Old World Monkeys, after that lineage had separated from the prosimians and New World Monkeys. Although some nonhuman primates continue to produce Theta-defensin peptides today, Homo sapiens and his gorilla, bonobo, and chimpanzee relatives do not. Their inability to do so reflects the common inheritance of defective Theta defensin genes which contain a premature stop codon that aborts translation. We can only speculate if this ancient genetic event has any contemporary relevance. However, it is noteworthy that synthetic retrocyclins can prevent the entry of HIV-1 and other viruses (e.g., HSV-2) into otherwise susceptible human target cells. Our studies suggest that their antiviral properties are intimately linked to an ability to bind carbohydrate epitopes displayed by viral and cell-surface glycoproteins involved in viral entry. The ability of retrocyclins to recognize and bind carbohydrate and glycan moieties is shared by Theta-defensins (RTDs) from rhesus monkeys and by several--but not all--human alpha-defensins. In addition to being the only cyclic peptides of animal origin, the lectin-like activity of Theta-defensins gives them the added distinction of being the smallest sugar-binding molecules of natural origin identified to date. This unusual combination makes Theta-defensins intriguing molecular prototypes that could be used to design novel carbohydrate-binding or antiviral agents. PMID- 15544533 TI - Microcin J25, from the macrocyclic to the lasso structure: implications for biosynthetic, evolutionary and biotechnological perspectives. AB - Microcin J25 (MccJ25) is a cyclic antibacterial peptide secreted by a fecal isolate of Escherichia coli. It exerts highly potent activity on Salmonella and Escherichia species. The microcin is recognized at the outer membrane of sensitive strains by the FhuA multifunctional protein, which belongs to the iron/siderophore receptor family, and inhibits bacterial transcription through binding to the RNA-polymerase beta' subunit. The mcjABCD genetic system carried by the wild type 50-kb pTUC100 plasmid contains four genes involved in MccJ25 production and immunity. MccJ25 results from the proteolytic cleavage of a 58 residue precursor at a specific Lys-Gly bond. The resulting mature peptide consists of 21 unmodified amino acids, mostly hydrophobic and includes a single dehydration. The initially described macrocyclic structure of MccJ25, which mostly relied on manual Edman sequencing of the thermolysin-cleaved form (t MccJ25), involved a head-to-tail cyclisation of the 21-residue precursor. This structure did not prove to be consistent with recent IT-MS CID experiments conducted either on the native microcin or on peptides resulting from acidic or enzymatic cleavages, which are in favour of an 8-residue ring followed by a 13 residue tail. Cyclisation thus occurs between the N-terminus (Gly1) and the Glu8 side chain carboxyl group. The solution three-dimensional structure shows threading of the tail into the ring, thus forming a highly stable lasso type structure. Such a structure was described previously for enzyme inhibitors from Actinobacteria and is consistent with the ability of MccJ25 to inhibit RNA polymerase. The lasso structure is discussed in terms of phylogenetical and biotechnological perspectives. PMID- 15544534 TI - The circular bacteriocins gassericin A and circularin A. AB - Gassericin A, a bacteriocin produced by Lactobacillus gasseri LA39, shows antibacterial activity against a number of Gram-positive food-borne pathogenic bacteria. Circularin A produced by Clostridium beijerinckii ATCC25752 is active against C. tyrobutyricum, a known cheese-spoilage bacterium. Both bacteriocins were purified to homogeneity from culture supernatants by reverse-phase chromatography and the subsequently determined amino acid sequences were used to clone the bacteriocin structural genes. Mature gassericin A and circularin A are class V circular bacteriocins comprised of 58 and 69 amino acid residues, respectively. Both bacteriocins are resistant to several peptidases and proteases, as are other cyclic bacteriocins. Heterologous expression of gassericin A in Escherichia coli was used to produce a non-cyclic mature peptide, which was shown to have a specific activity 173-fold lower than the circular molecule. The minimal region for production and secretion of active circularin A is comprised of five genes, as was deduced by heterologous gene expression in Enterococcus faecalis. Gassericin A and circularin A have limited mutual similarity in their primary sequences. Unlike most bacteriocins, including gassericin A, circularin A has a three-amino-acid-leader sequence. PMID- 15544535 TI - Peptide AS-48: prototype of a new class of cyclic bacteriocins. AB - After the discovery of bacteriocin AS-48, a 70-residue cyclic peptide produced by Enterococcus faecalis subsp. liquefaciens, some naturally-occurring cyclic proteins from bacteria have been reported. AS-48 is encoded by the 68-kb pheromone-responsive plasmid pMB2, and the gene cluster involved in production and immunity has been identified and sequenced. This peptide exerts a bactericidal action on sensitive cells (most of the Gram-positive and some Gram negative bacteria). Its target is the cytoplasmic membrane, in which it opens pores, leading to the dissipation of the proton motive force and cell death, a mechanism similar to that proposed for the action of defensins or, most generally, cationic antibacterial peptides. This fact, together with its remarkable stability and solubility over a wide pH range, suggest that this bacteriocin could be a good candidate as a natural food preservative. The amino acid composition of purified AS-48 shows the absence of modified or dehydrated residues, making it clearly different from lantibiotics. Bacteriocin AS-48 also differs from defensins in that it does not contain cysteines and consequently no disulfide bridges, which makes is high stability even more remarkable. Composition analysis of AS-48 shows a high proportion of basic to acidic amino acids, conferring to this peptide a strong basic character, with an isoelectric point close to 10.5. Determination of the AS-48 structural gene DNA sequence, together with the sequences of AS-48 protease digestion fragments and mass spectrometry determinations, allowed us to determine unambiguously the cyclic structure of the molecule, being the first example of a posttranslational modification in which a cyclic structure arises from a "head-to-tail" linkage. We have solved the three-dimensional structure of AS-48 in solution, and it consists of a globular arrangement of five alpha-helices enclosing a compact hydrophobic core. Interestingly, the head-to-tail peptide link between Trp-70 and Met-1 lies in the middle of alpha-helix 5, which is shown to have a pronounced effect on the stability of the three-dimensional structure. Analysis of structure-function relationship allowed us to propose models to understand the aspects of the molecular function of AS-48. The purpose of this work is to review recent developments in our understanding about the biochemical and biological characteristics and structure of this unusual type of bacteriocin. PMID- 15544536 TI - Protein circlets as sex pilus subunits. AB - The largest circular protein structures discovered define a class of transfer proteins acting in bacterial conjugation and type IV secretion. Proteins ranging from 73 to 78 residues with head-to-tail peptide bonds constitute the major subunit of conjugative pili of some type IV secretion systems. Their plasmid encoded precursors are enzymatically processed and cyclized before being assembled into pili. These extra-cellular surface filaments mediate physical contact between donor and recipient cell or pathogen and host cell. Pili are essential prerequisites for DNA and protein transfer. A membrane-bound signal peptidase-like enzyme is responsible for the circularization reaction. Site directed mutagenesis and mass spectrometry has been used extensively to unravel the mechanism of the enzyme-substrate interaction of the pilin maturation process. PMID- 15544537 TI - The discovery and potential of N-sulfonylated dipeptide VLA-4 antagonists. AB - Through a directed screening of a combinatorial library containing carboxylic acids, N-sulfonylated dipeptides were identified as leads in the Merck Research Laboratories VLA-4 antagonist program. Further optimization quickly identified subnanomolar compounds with varying degrees of specificity over the related integrin alpha4beta7. Various metabolic liabilities were identified and addressed. However, the pharmacokinetic properties of nearly all compounds in this class were unacceptable. Other leads were identified with apparent good oral bioavailability, but these were generally associated with very high plasma protein binding and a loss of potency. The mechanism of high plasma clearance was identified in the rat as the organic acid transporter, mrp-2. Compounds were identified that were not substrates of mrp-2, but they still suffered from poor oral bioavailability. Finally, a shift in strategy to identifying VLA-4 antagonists that would be suitable as candidates for inhalation therapy resulted in the preparation of compounds with exception tight binding properties. These compounds were superior to BIO-1211 in the ovalbumin-sensitized mouse model of eosinophil trafficking to the lung. One particular compound had an exceptionally long off-rate with a KD < or = 2 pM. The evolution of the structure activity relationships in our laboratories and strategies for improving potencies and pharmacokinetic profiles are the subject of this review. PMID- 15544538 TI - The identification and optimization of orally efficacious, small molecule VLA-4 antagonists. AB - The identification of orally active, small molecule antagonists of the alpha4beta1 integrin, VLA-4, could lead to therapeutic agents with utility in a number of clinical settings, including asthma, multiple sclerosis and IBD. Starting from CDR3 sequences conserved among neutralizing alpha4 antibodies, peptides were identified that antagonized VLA-4 mediated adhesion in vitro. Through a series of structural modifications, these peptides evolved into small molecules that exhibited high potency and selectivity for VLA-4 in cell adhesion assays. Finally, through the optimization of physical and pharmacokinetic properties, compounds were identified that exhibited oral activity in animal models of asthma and multiple sclerosis. PMID- 15544539 TI - Therapeutic antagonists and the conformational regulation of the beta2 integrins. AB - The beta2 integrins are validated therapeutic targets for inflammatory disorders. Two distinct mechanistic classes of small molecule inhibitors, termed alpha I allosteric and alpha/beta I-like allosteric antagonist, have recently been developed. The alpha I allosteric antagonists bind underneath the C-terminal helix of the I domain and stabilize the I domain in the inactive closed conformation. By contrast, the alpha/beta I-like allosteric antagonists bind to the beta2 I-like domain MIDAS and disrupt conformational signal transmission between the I and the I-like domain, leaving the I domain in a default inactive form. Furthermore, the two classes of the antagonists have opposite effects on integrin conformation; the alpha I allosteric antagonists stabilize the bent conformation, whereas the alpha/beta I-like allosteric antagonists induce the extended conformation with inactive I domain. The small molecule antagonists to the beta2 integrin highlight the importance of the structural linkages within and between integrin domains for transmission of the conformational signals and regulation of the overall conformation. PMID- 15544540 TI - Rational design of potent and selective VLA-4 inhibitors and their utility in the treatment of asthma. AB - Asthma, a chronic inflammatory disease of the airways, is a significant burden on our healthcare system. There is high unmet need for treatments directed towards the underlying causes of the disease. The cell surface integrin VLA-4 (very late antigen-4; alpha4beta1; CD49d/CD29) plays an important role in the trafficking of white blood cells to sites of inflammation and represents an exciting target for the development of novel anti-inflammatory drugs for the treatment of asthma. Here, we review our efforts to use rational design to identify potent, selective inhibitors of VLA-4. We describe the discovery of a series of potent VLA-4 inhibitors through the addition of a novel N-terminal organic cap to a tetrapeptide VLA-4 binding motif 4-((N'-2-methylphenyl)uriedo)phenylacetyl-Leu Asp-Val-Pro ; Kd = 70 pM), and rationalize their structure-activity relationships using 3D-QSAR. Also, we show our rational peptidomimetic design strategy using "template hopping" from the gpIIb/IIIa integrin antagonist field, and also a novel virtual screening strategy. Two series have been developed, one that has high selectivity for the activated over the non-activated state of the receptor, and the other which is non-selective inhibiting both activated and non-activated VLA-4. Both series are highly selective for VLA-4 versus against other integrin family members. These inhibitors show promise in the treatment of asthma, based upon efficacy in a sheep model of asthma, where they inhibit both the early and late-phase responses to asthma and also block hypersensitivity. PMID- 15544541 TI - The discovery of VLA-4 antagonists. AB - Starting with a cyclic peptide of moderate potency as a VLA-4 antagonist, highly potent and conformationally defined cyclic peptides were developed incorporating a constrained tyrosine and an achiral Asp-Pro spacer. N-Acyl phenylalanine derivatives were also discovered to have VLA-4 antagonist activity. During the course of development of this series, we found that the active acylphenylalanines mimic the pharmacophores present in the cyclic peptides and hypothesized that they bind to the same site on VLA-4. This insight guided our optimization strategy. Based on the emerging SAR, as well as insights from the recent X-ray crystal structure of the integrin alphavbeta3 bound to a RGD containing cyclic peptide, we propose a binding model for these compounds. PMID- 15544542 TI - Biologically active dietary peptides. AB - A large variety of peptides are generated in the gut lumen during normal digestion of dietary proteins. Large quantities of small peptides (ie. dipeptides and tripeptides) are absorbed through the gut mucosa and represent the primary mechanism for absorption of dietary nitrogen. However, larger peptide fragments are also absorbed with absorption decreasing with increasing chain length. Many of these dietary peptides have been shown to have biologic activity and many are active in microgram quantities. These peptides may modulate neural, endocrine, and immune function. In this report, we review normal protein digestion and absorption. We then discuss the biological actions of the amino acids arginine and glutamine and the biologic actions of a variety of dietary derived peptides. We concentrate on the immune effects of these peptides. We illustrate the potency of dietary peptides with a discussion of the cardiovascular effects of carnosine. We also review biologic effects of different protein sources, which generate different peptide profiles during digestion. The implications of dietary peptides for modulation of disease are discussed. PMID- 15544543 TI - Cellular and physiological effects of arginine. AB - Arginine is a semi-essential amino acid that is required during periods of maximal growth, severe stress, and injury. Arginine is a substrate for protein synthesis but also modulates cellular biochemical functions via conversion to a number of biologically active compounds. Arginine is utilized by a vast variety of metabolic pathways that produce a variety of biologically active compounds such as nitric oxide, creatine phosphate, agmatine, polyamines, ornithine, and citrulline. Arginine supply is primarily regulated by two enzyme systems: arginase (part of the urea cycle) and nitric oxide synthase. Arginine has many effects in the body that include modulation of immune function, wound healing, hormone secretion, vascular tone, insulin sensitivity, and endothelial function. Arginine mediates its effects via nitric oxide independent and dependent pathways. Nitric oxide modulates many cellular functions that include vascular tone, expression of adhesion molecules, leukocyte adhesion, and platelet aggregation. Arginine modulates the development of atherosclerotic cardiovascular disease, improves immune function in healthy and ill patients, stimulates wound healing in healthy and ill patients, and modulates carcinogenesis and tumor growth. Thus, arginine is a biologically active dietary compound with numerous physiologic and pharmacological activities. PMID- 15544544 TI - Regulation of the cellular and physiological effects of glutamine. AB - Glutamine is the most abundant amino acid in humans and possesses many functions in the body. It is the major transporter of amino-nitrogen between cells and an important fuel source for rapidly dividing cells such as cells of the immune and gastrointestinal systems. It is important in the synthesis of nucleic acids, glutathione, citrulline, arginine, gamma aminobutyric acid, and glucose. It is important for growth, gastrointestinal integrity, acid-base homeostasis, and optimal immune function. The regulation of glutamine levels in cells via glutaminase and glutamine synthetase is discussed. The cellular and physiologic effects of glutamine upon the central nervous system, gastrointestinal function, during metabolic support, and following tissue injury and critical illness is also discussed. PMID- 15544545 TI - Cellular and physiological effects of short-chain fatty acids. AB - Short chain fatty acids (SCFA) have multiple in vivo and in vitro effects including cell cycle arrest and induction of protein synthesis, differentiation, and apoptosis. Butyrate or derivatives have promise for treating inflammatory bowel disease and colon cancer. Genomic and proteomic analysis have generated new knowledge on function and applications for SCFA. PMID- 15544546 TI - Cellular and physiological effects of medium-chain triglycerides. AB - From a nutritional standpoint, saturated triglycerides with a medium (6 to 12) carbon chain length (MCT) have traditionally been regarded as biologically inert substances, merely serving as a source of fuel calories that is relatively easily accessible for metabolic breakdown compared with long chain triglycerides (LCT). This quality of MCT has been shown to offer both benefits and risks depending on the clinical situation, with potential positive effects on protein metabolism in some studies on one side, and an increased risk for ketogenesis and metabolic acidosis on the other. At another level, studies regarding lipid effects of MCT on the immune system, as with LCT, so far have yielded equivocal results, although there is a recent experimental evidence to suggest that MCT possess immune modulating properties and should in fact be regarded as bioactive mediators. Most of this information comes from studies where effects of MCT have been compared with those of LCT in lipid emulsions, as part of parenteral (intravenous) nutrition formulations. Unfortunately, the relevance of these observations for clinical practice remains largely unclear because adequately powered trials that clearly point out the position of MCT in relation to structurally different lipids have not been performed. In the present paper we review the experimental and clinical evidence for cellular and physiological effects of nutritional MCT. In addition, studies describing possible mechanisms behind the observed effects of MCT will be discussed. PMID- 15544547 TI - Omega 3-fatty acids: health benefits and cellular mechanisms of action. AB - Epidemiological evidence has established that ingestion of long-chain polyunsaturated omega-3 fatty acids (omega-3 PUFAs), abundant in fish oils, have profound effects on many human disorders and diseases, including cardiovascular disease and cancer. Here we briefly review the dietary recommendations and the food sources that are naturally enriched by these fatty acids. There are also a number of products including eggs, bread, and cereals available to supplement omega-3 fatty acid dietary intake. Some of these supplements are proposed to aid different pathological conditions. While the beneficial effects of omega-3 fatty acids can no longer be doubted, their molecular mechanism of action remains elusive. Without question, the action of omega-3 fatty acids is complex and involves a number of integrated signaling pathways. This review focuses on one of the possible cellular mechanisms by which the omega-3 PUFAs, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), may function. Studies with cancer cells suggest that DHA induces cell cycle arrest and apoptosis by activating protein phosphatases, leading to dephosphorylation of retinoblastoma protein (pRB). Protein phosphatases are also involved with the protein Bcl2, which regulates the release of cytochrome c from mitochondria, and eventually, activation of the apoptotic enzyme caspase 3. PMID- 15544548 TI - Cellular and physiological effects of Ganoderma lucidum (Reishi). AB - In Asia, a variety of dietary products have been used for centuries as popular remedies to prevent or treat different diseases. A large number of herbs and extracts from medicinal mushrooms are used for the treatment of diseases. Mushrooms such as Ganoderma lucidum (Reishi), Lentinus edodes (Shiitake), Grifola frondosa (Maitake), Hericium erinaceum (Yamabushitake), and Inonotus obliquus (Chaga) have been collected and consumed in China, Korea, and Japan for centuries. Until recently, these mushrooms were largely unknown in the West and were considered 'fungi' without any nutritional value. However, most mushrooms are rich in vitamins, fiber, and amino acids and low in fat, cholesterol, and calories. These mushrooms contain a large variety of biologically active polysaccharides with immunostimulatory properties, which contribute to their anticancer effects. Furthermore, other bioactive substances, including triterpenes, proteins, lipids, cerebrosides, and phenols, have been identified and characterized in medicinal mushrooms. This review summarizes the biological effects of Ganoderma lucidum upon specific signaling molecules and pathways, which are responsible for its therapeutic effects. PMID- 15544549 TI - Cellular and physiological effects of soy flavonoids. AB - Recent experimental and epidemiological studies have provided convincing evidence for a variety of health benefits derived from the consumption of soy and soy food products. For example, soy isoflavones are felt to protect against different cancers, cardiovascular disease, and bone loss. Many studies have demonstrated the effect of soy isoflavones on specific target molecules and signaling pathways, including but not limited to, cell proliferation and differentiation, cell cycle regulation, apoptosis, angiogenesis, cell adhesion and migration, metastasis, and activity of different enzymes. Isoflavones also share structural homologies with estrogens and are therefore classified as phytoestrogens with weak estrogenic properties. Since isoflavones bind to estrogen receptors (ER alpha and ER beta), they are considered to be possible estrogen receptor modulators. However, isoflavones can also exert biological effects independent of their phytoestrogenic activities. Recent studies suggest beneficial health effects of soy and recommend increasing the intake of isoflavone-rich soy protein to the level of intake commonly used in Asian countries. PMID- 15544550 TI - Cellular and physiological effects of probiotics and prebiotics. AB - We review the present knowledge on the biological mechanisms of action of probiotics and prebiotics. They include direct effects in the intestinal lumen or on intestinal or immune cells, and indirect mechanisms through modulation of the endogenous microflora (composition or functions such as butyrate production) or of the immune system. PMID- 15544551 TI - Intervention of hepatic glucose production. Small molecule regulators of potential targets for Type 2 diabetes therapy. AB - Excessive hepatic glucose production is thought to be a major contributor to the type 2 diabetic state. Drug discovery efforts have yielded small synthetic inhibitors for gluconeogenic and glycogenic regulators of this pathway. The most advanced targets are outlined in this mini-review and include: the glucocorticoid receptor, 11 beta-hydroxysteroid dehydrogenase type 1, fructose 1,6 bisphosphatase, the glucagon receptor, glycogen phosphorylase, glycogen synthase kinase-3, and glucose-6-phosphatase. PMID- 15544552 TI - Development of radioligands for in vivo imaging of GABA(A)-benzodiazepine receptors. AB - Changes in the biochemical integrity and function of the GABA(A)-benzodiazepine receptor (BZR) complex have been implicated in various neurological and psychiatric disorders. The development of specific radioligands for the GABA(A) BZR have not only contributed to the elucidation of the receptor's biochemical functions, but also provided a means by which these changes are correlated to disease states when studied with the functional imaging modalities of positron emission tomography (PET) and single photon emission computed tomography (SPECT). PMID- 15544553 TI - Dual acting antihistaminergic agents. AB - Histamine is a primary mediator in allergic response and acts in concert with other agents to impact disease progression. Respiratory disorders such as asthma, rhinitis and dermatological conditions such as urticaria involve histamine along with other mediators. An antihistamine that possesses an additional property of counteracting the effects mediated by these other mediators should offer some therapeutic benefit over a selective antihistaminergic agent. PMID- 15544554 TI - Structure-activity relationships of histamine H1-receptor agonists. AB - Significant progress in the development of potent and selective histamine H1 receptor agonists has been achieved since 1990. Optimisation of the class of 2 phenylhistamines has furnished 2-[3-(trifluoromethyl)phenyl]histamine and its Nalpha-methyl derivative. The discovery of histaprodifen (2-[2-(3,3 diphenylpropyl)-1H-imidazol-4-yl]ethanamine) and the novel lead compound suprahistaprodifen (Nalpha-2-[(1H-imidazol-4-yl)ethyl]histaprodifen) represents additional milestones in the H1-receptor agonist field. PMID- 15544555 TI - Structure-activity relationships of histamine H2 receptor ligands. AB - Recent research on histamine H2 receptor agonists was focused on quantitative structure-activity relationships and receptor models explaining the activity of imidazolylpropylguanidines. Their selectivity for guinea pig vs. human isoforms was investigated using H2 receptor-Gsalpha fusion proteins and attributed to amino acid differences in transmembrane domains 1 and 7. New antagonists result from approaches to improve pharmacokinetic properties and to design hybrid drugs which additionally have gastroprotective or anti H. pylori activity. PMID- 15544556 TI - Histamine H3 receptor agonists. AB - The SAR of H3 ligands has been difficult to evaluate because of species differences, multiple isoforms and constitutive activity, among other complicating factors. A review is given of the sometimes-conflicting affinity, activity and efficacy data of H3 agonists that has been described in literature to date. PMID- 15544557 TI - Medicinal chemical and pharmacological aspects of imidazole-containing histamine H3 receptor antagonists. AB - The first antagonists known for the histamine H3 receptor were mono-substituted imidazole-containing compounds like thioperamide. Meanwhile numerous novel leads have been developed possessing improved affinities, selectivities, specificities, and pharmacokinetic properties. Scope and limitations of this promising class are discussed concerning their structure-activity relationships as well as pharmacological and potential therapeutic aspects. PMID- 15544558 TI - Medicinal chemistry and biological properties of non-imidazole histamine H3 antagonists. AB - The H3 receptor is prominently expressed in neuronal tissues, and H3 antagonists have been proposed as drugs with benefits in disorders of cognition, attention, pain, allergic rhinitis, and obesity. The structure-activity relationships (SAR) of various classes of non-imidazole H3 antagonists are reviewed, along with highlights of functional efficacy in tissue-based and animal disease models. PMID- 15544559 TI - The histamine H4 receptor and potential therapeutic uses for H4 ligands. AB - Histamine is a biogenic amine that plays a host of physiological roles and the three major functions for histamine have been largely defined by the activity of three receptors. The inflammatory wheal and flare response is driven by the H1 receptor [1]. The H2 receptor controls gastric acid secretion in the gut [2]. The H3 receptor is involved in neurotransmitter release in the central nervous system [3]. The recent discovery of the histamine H4 receptor by several groups has lead to the re-evaluation of the physiological role for histamine. PMID- 15544560 TI - Recent advances in the medicinal chemistry of alpha-aminoboronic acids, amine carboxyboranes and their derivatives. AB - This article describes recent developments in the synthesis and biological activity of alpha-aminoboronic acids, amine-carboxyboranes and their derivatives as potential therapeutic agents. alpha-Amino acid analogues are of considerable interest as inhibitors of enzymes involved in amino acid and peptide metabolism. In particular, alpha-amino alkylphosphonic acids and alpha-amino alkylboronic acids, in which the carboxyl group of amino acids is replaced by a phosphonic acid or boronic acid function, respectively, constitute a unique class of amino acid mimics from which a number of potent enzyme inhibitors have been synthesized. The inhibitory activity mainly stems from the fact that the tetrahedral phosphonic moiety or the tetrahedral adduct of electrophilic boronic acid is a good mimic of the putative tetrahedral transition state or intermediate encountered in the enzymatic hydrolysis or formation of peptides. Since the peptide hydrolysis and formation invariably involves the tetrahedral high energy species in the course of the reaction, these amino acid mimics serve as a general key element for inhibitors of a broad spectrum of proteases and peptide ligases. Serine protease inhibitors provide promising compounds having a P site binding moiety and a boronic acid chelating moiety. The compounds have been shown to have high inhibitory activity. PMID- 15544561 TI - The zinc metallopeptidase family: new faces, new functions. PMID- 15544562 TI - The intracellular distribution and secretion of endopeptidases 24.15 (EC 3.4.24.15) and 24.16 (EC 3.4.24.16). AB - Endopeptidase 24.15 (EC 3.4.24.15; EP24.15) and endopeptidase 24.16 (EC 3.4.24.16; EP24.16) are enzymes involved in general peptide metabolism in mammalian cells and tissues. This review will focus on morphological and biochemical aspects related to the subcellular distribution and secretion of these homologous enzymes in the central nervous system. These are important issues for a better understanding of the functions of EP24.15 and EP24.16 within neuroendocrine systems. PMID- 15544563 TI - Secretase-mediated cell surface shedding of the angiotensin-converting enzyme. AB - Angiotensin-converting enzyme (ACE) is an example of a membrane-bound protein, which is shed from the cell surface in a soluble form by a post-translational proteolytic cleavage event involving a secretase. The secretase cleavage site in somatic ACE has been mapped to Arg-1203/Ser-1204, 24 residues proximal to the membrane-anchoring domain and the ADAM ('a disintegrin and metalloprotease') family of proteins may be involved in ACE shedding. PMID- 15544564 TI - Hemodynamic regulation of metallopeptidases within the vasculature. AB - Hemodynamic forces associated with blood flow play a vital role in the endothelial regulation of vascular tone, remodeling and the initiation and progression of vascular diseases such as atherosclerosis and hypertension. Crucial elements in endothelium-mediated events within the blood vessel are bioactive peptide signals and their associated hydrolytic enzymes. This review examines the relationship between hemodynamic forces such as shear stress and cyclic strain, and an important group of peptide-degrading enzymes within the endothelium, the thermolysin-like zinc metallopeptidases. PMID- 15544565 TI - Metalloproteinase-mediated shedding of heparin-binding EGF-like growth factor and its pathophysiological roles. AB - Heparin-binding EGF-like growth factor (HB-EGF) exists as a membrane-anchored form (proHB-EGF) and as its soluble cleaved product (sHB-EGF). The conversion (ectodomain shedding) of proHB-EGF to sHB-EGF is tightly regulated by specific metalloproteinases. Ectodomain shedding plays a central role in GPCR-mediated EGFR transactivation. Antagonizing metalloproteinases can inhibit EGFR transactivation and might be of therapeutic value, for example in cardiac hypertrophy, skin remodeling and tumor growth. PMID- 15544566 TI - DINE (damage induced neuronal endopeptidase). AB - A unique central nervous system (CNS)-specific metalloprotease, DINE/ECEL1 (damage induced neuronal endopeptidase/ endothelin converting enzyme-like 1), has recently been added to the M13/neprilysin (NEP) family. This enzyme was identified by two groups independently using different approaches. In this review, we introduce the characteristics of DINE/ECEL1 and focus on the mechanism underlying the transcriptional regulation of DINE in response to neuronal injury. PMID- 15544567 TI - Endothelin converting enzyme-2: a processing enzyme involved in the generation of novel neuropeptides. AB - Members of several metalloprotease families have been proposed to be involved in non-classical processing of neuroendocrine precursors. Among them, endothelin converting enzyme-2 (ECE-2) is a good candidate since it exhibits a neuroendocrine distribution, intracellular subcellular localization, and an acidic pH optimum. The enzyme is able to generate a number of biologically active peptides from peptide intermediates, suggesting an important role for this enzyme in the biosynthesis of regulatory peptides. These results are consistent with an important role for ECE-2 in the processing of regulatory peptides at non classical sites. PMID- 15544568 TI - The role of neuropeptide processing enzymes in endocrine (prostate) cancer: EC 3.4.24.15 (EP24.15). AB - The zinc metalloendopeptidase EC3.4.24.15 [EP24.15, thimet oligopeptidase], a neuropeptide processing enzyme, is central to the formation and degradation of many bioactive peptides in the neural proteome, and is highly expressed in normal prostate. EP24.15 actions are increased in androgen-dependent prostate cancer compared to androgen-independent; augmented by androgen treatment, and inhibited by clinical GnRH analogs. The "neural" prostate includes: neuropeptides, cognate receptors and processing enzymes regulating signaling of peptide-mediated neural inputs. PMID- 15544569 TI - Neprilysin 2: a novel messenger peptide-inactivating metalloprotease. AB - Neprilysin 2 is a recently identified glycoprotein displaying the highest degree of sequence identity with neprilysin (EC 3.4.24.11), the prototypical member of the M13 family of zinc-dependent metalloproteases. Whereas neprilysin has been shown to be involved in the inactivation of endogenous messenger peptides, like enkephalins and tachykinins, the true physiological functions of neprilysin 2 remain unknown. PMID- 15544570 TI - Membrane bound members of the M1 family: more than aminopeptidases. AB - In mammals the M1 aminopeptidase family consists of nine different proteins, five of which are integral membrane proteins. The aminopeptidases are defined by two motifs in the catalytic domain; a zinc binding motif HEXXH-(X18)-E and an exopeptidase motif GXMEN. Aminopeptidases of this family are able to cleave a broad range of peptides down to only to a single peptide. This ability to either generate or degrade active peptide hormones is the focus of this review. In addition to their capacity to degrade a range of peptides a number of these aminopeptidases have novel functions that impact on cell signalling and will be discussed. PMID- 15544571 TI - Nardilysin, a basic residues specific metallopeptidase that mediates cell migration and proliferation. AB - Nardilysin (NRDc), a metallopeptidase of the M16 family, presents, in vitro, cleavage specificity for basic residues. Depending on the cell type, it is cytoplasmic, exported or cell surface associated. As a new receptor for heparin binding EGF-like growth factor (HB-EGF), NRDc was recently shown to be involved in cellular migration and proliferation. Since for those processes its enzymatic activity is not required, it is now evident that nardilysin fulfills at least two distinct functions, i.e. an HB-EGF modulator and a peptidase. PMID- 15544572 TI - Short-term blood pressure and heart rate variability in congenital central hypoventilation syndrome (Ondine's curse). AB - The effect of CCHS (congenital central hypoventilation syndrome, or Ondine's curse) on short-term BP (blood pressure) and HR (heart rate) variability was evaluated in 16-year-old subjects presenting a form of CCHS requiring night ventilatory assistance. The 12 patients were compared with 12 age- and gender matched healthy volunteers. Recordings were obtained during daytime while the subjects were breathing spontaneously. Continuous BP was measured with a Finapres device in the supine, head-up tilt and standing positions. The manoeuvre of actively standing was also analysed. HR levels were elevated in CCHS subjects at supine rest (+23%) with a reduced HR overall variability (-88%). The low- and high-frequency components of HR variability were affected. BP levels were preserved at rest, but the manoeuvres demonstrated a limited capacity to elevate BP. There was no overshoot in BP during the manoeuvre of actively standing, and steady standing BP levels in patients were not higher than supine BP levels as usually observed in healthy controls. The spontaneous baroreflex sensitivity estimated using the sequence technique or the cross-spectral analysis fell in the patients to approx. one-third of the sensitivity estimated in the healthy controls whatever the position. This cardiovascular profile suggests a predominant vagal dysfunction with signs of vagal withdrawal and baroreflex failure, and relative preservation of the cardiac and vascular sympathetic function. It is likely that the impaired ontogeny of the visceral reflexes, considered now to cause CCHS syndrome, includes the baroreceptive pathway and mainly its vagal component. PMID- 15544573 TI - New insights into amino acid metabolism, beta-cell function and diabetes. AB - Specific amino acids are now known to acutely and chronically regulate insulin secretion from pancreatic beta-cells in vivo and in vitro. Understanding the molecular mechanisms by which amino acids regulate insulin secretion may identify novel targets for future diabetes therapies. Mitochondrial metabolism is crucial for the coupling of amino acid and glucose recognition to the exocytosis of the insulin granules. This is illustrated by in vitro and in vivo observations discussed in the present review. Mitochondria generate ATP, which is the main coupling factor in insulin secretion; however, the subsequent Ca2+ signal in the cytosol is necessary, but not sufficient, for full development of sustained insulin secretion. Hence mitochondria generate ATP and other coupling factors serving as fuel sensors for the control of the exocytotic process. Numerous studies have sought to identify the factors that mediate the amplifying pathway over the Ca2+ signal in nutrient-stimulated insulin secretion. Predominantly, these factors are nucleotides (GTP, ATP, cAMP and NADPH), although metabolites have also been proposed, such as long-chain acyl-CoA derivatives and the key amino acid glutamate. This scenario highlights further the importance of the key enzymes or transporters, glutamate dehydrogenase, the aspartate and alanine aminotransferases and the malate/aspartate shuttle, in the control of insulin secretion. Therefore amino acids may play a direct or indirect (via generation of putative messengers of mitochondrial origin) role in insulin secretion. PMID- 15544574 TI - Generation and function of astroglial lipoproteins from Niemann-Pick type C1 deficient mice. AB - NPC (Niemann-Pick type C) disease is a progressive neurological disorder characterized by defects in intracellular cholesterol trafficking, accumulation of cholesterol in the endosomal system and impaired cholesterol homoeostasis. Although these alterations appear to occur in all NPC1-deficient cell types, the consequences are most profound in the nervous system. Since glial cells are important mediators of brain cholesterol homoeostasis, we proposed that defective generation and/or function of lipoproteins released by glia might contribute to the neurological abnormalities associated with NPC disease. We found that, as in other cell types, Npc1-/- glia accumulate cholesterol intracellularly. We hypothesized that this sequestration of cholesterol in glia might restrict the availability of cholesterol for lipoprotein production. Cerebellar astroglia were cultured from a murine model of NPC disease to compare the lipoproteins generated by these cells and wild-type glia. The experiments demonstrate that the amount of cholesterol in glia-conditioned medium is not reduced by NPC1 deficiency. Similarly, cholesterol efflux to apo (apolipoprotein) A1 or glial expression of the transporter ATP-binding-cassette transporter A1 was not decreased by NPC1 deficiency. In addition, the ratio of apo E:cholesterol and the density distribution of lipoproteins in Npc1-/- and Npc1+/+ glia-conditioned medium are indistinguishable. Importantly, in a functional assay, apo E-containing lipoproteins generated by Npc1-/- and Npc1+/+ glia each stimulate axonal elongation of neurons by approx. 35%. On the basis of these observations, we speculate that the neuropathology characteristic of NPC disease can quite probably be ascribed to impaired processes within neurons in the brain rather than defective lipoprotein production by astroglia. PMID- 15544575 TI - The individuality of mice. AB - Mutant mice simulating human CNS disorders are used as models for therapeutic drug development. Drug evaluation requires a coherent correlation between behavioral phenotype and drug status. Variations in behavioral responses could mask such correlations, a problem highlighted by the three-site studies of Crabbe et al. (1999) and Wahlsten et al. (2003a). Factors contributing to variation are considered, focusing on differences between individual animals. Genetic differences due to minisatellite variation suggest that each mouse is genetically distinct. Effects during gestation, including maternal stress, influence later life behavior; while endocrine exchanges between fetus and parent, and between male and female fetuses dependent on intrauterine position, also contribute. Pre and perinatal nutrition and maternal attention also play a role. In adults, endocrine cyclicity in females is a recognized source of behavioral diversity. Notably, there is increasing recognition that groups of wild and laboratory mice have complex social structures, illustrated through consideration of Crowcroft (1966). Dominance status can markedly modify behavior in test paradigms addressing anxiety, locomotion and aggressiveness, to an extent comparable to mutation or drug status. Understanding how such effects amplify the behavioral spectrum displayed by otherwise identical animals will improve testing. PMID- 15544576 TI - Association of tryptophan hydroxylase gene polymorphism with depression, anxiety and comorbid depression and anxiety in a population-based sample of postpartum Taiwanese women. AB - Depression and anxiety disorders often coexist clinically and both are known to have a genetic basis, but the mode of inheritance is too complicated to be determined so far. Serotonin is the biogenic amine neurotransmitter most commonly associated with depression and anxiety. Since tryptophan hydroxylase (TPH1) is the rate-limiting enzyme in serotonin biosynthesis, its role in the pathophysiology of these psychiatric diseases has been intensively studied. In this study, we examined whether polymorphism of the TPH1 gene is related to the etiology of major depression, anxiety and comorbid depression and anxiety. Five single nucleoside polymorphisms of the TPH1 gene were studied in a population based sample of postpartum Taiwanese women consisting of 120 subjects with depression or/and anxiety and 86 matched normal controls. A significant difference (P = 0.0107) in genotype frequency for the T27224C polymorphism was found between the comorbid and normal groups, and risk analysis showed that the C allele conferred a strong protective effect (odds ratio = 0.27; 95% confident interval = 0.11-0.7). Three-allele haplotypes involving T27224C polymorphism were constructed and haplotype associations between particular haplotype combinations and various diseases identified. However, the associations were weak and the overall haplotype frequency profiles in all groups were similar. The results suggest that depression, anxiety, and comorbid depression and anxiety disorders may have related etiologies. In addition, this study suggests that the TPH1 gene might play a role in the pathogenesis of these closely related disorders. PMID- 15544577 TI - A phenotypic and molecular characterization of the fmr1-tm1Cgr fragile X mouse. AB - Fragile X Syndrome is the most common form of inherited mental retardation. It is also known for having a substantial behavioral morbidity, including autistic features. In humans, Fragile X Syndrome is almost always caused by inactivation of the X-linked FMR1 gene. A single knockout mouse model, fmr1-tm1Cgr, exists. In this report we further characterize the cognitive and behavioral phenotype of the fmr1-tm1Cgr Fragile X mouse through the use of F1 hybrid mice derived from two inbred strains (FVB/NJ and C57BL/6J). Use of F1 hybrids allows focus on the effects of the fmr1-tm1Cgr allele with reduced influence from recessive alleles present in the parental inbred strains. We find that the cognitive phenotype of fmr1-tm1Cgr mice, including measures of working memory and learning set formation that are known to be seriously impacted in humans with Fragile X Syndrome, are essentially normal. Further testing of inbred strains supports this conclusion. Thus, any fmr1-tm1Cgr cognitive deficit is surprisingly mild or absent. There is, however, clear support presented for a robust audiogenic seizure phenotype in all strains tested, as well as increased entries into the center of an open field. Finally, a molecular examination of the fmr1-tm1Cgr mouse shows that, contrary to common belief, it is not a molecular null. Implications of this finding for interpretation of the phenotype are discussed. PMID- 15544578 TI - Convergent analysis of cDNA and short oligomer microarrays, mouse null mutants and bioinformatics resources to study complex traits. AB - Gene expression data sets have recently been exploited to study genetic factors that modulate complex traits. However, it has been challenging to establish a direct link between variation in patterns of gene expression and variation in higher order traits such as neuropharmacological responses and patterns of behavior. Here we illustrate an approach that combines gene expression data with new bioinformatics resources to discover genes that potentially modulate behavior. We have exploited three complementary genetic models to obtain convergent evidence that differential expression of a subset of genes and molecular pathways influences ethanol-induced conditioned taste aversion (CTA). As a first step, cDNA microarrays were used to compare gene expression profiles of two null mutant mouse lines with difference in ethanol-induced aversion. Mice lacking a functional copy of G protein-gated potassium channel subunit 2 (Girk2) show a decrease in the aversive effects of ethanol, whereas preproenkephalin (Penk) null mutant mice show the opposite response. We hypothesize that these behavioral differences are generated in part by alterations in expression downstream of the null alleles. We then exploited the WebQTL databases to examine the genetic covariance between mRNA expression levels and measurements of ethanol induced CTA in BXD recombinant inbred (RI) strains. Finally, we identified a subset of genes and functional groups associated with ethanol-induced CTA in both null mutant lines and BXD RI strains. Collectively, these approaches highlight the phosphatidylinositol signaling pathway and identify several genes including protein kinase C beta isoform and preproenkephalin in regulation of ethanol- induced conditioned taste aversion. Our results point to the increasing potential of the convergent approach and biological databases to investigate genetic mechanisms of complex traits. PMID- 15544579 TI - Building research capacity in nursing. PMID- 15544580 TI - Suffering related to health care: a study of breast cancer patients' experiences. AB - A previous study indicated that patient narratives include experiences of suffering caused or increased by health-care encounters. The aim of this study was to interpret and understand the meaning of patients' experiences of suffering related to health care from an ethical, existential and ontological standpoint. Sixteen women with breast cancer in Sweden and Finland took part in qualitative interviews analysed with a hermeneutic, interpretive approach. The outcome showed that suffering related to health care is a complex phenomenon and constitutes an ethical challenge to health-care personnel. The women's experiences of suffering related to health care tended to be of similar seriousness as their experiences of suffering in relation to having cancer. In an ethical, existential and ontological sense, suffering related to health care is basically a matter of neglect and uncaring where the patient's existential suffering is not seen and she is not viewed as a whole human being. PMID- 15544581 TI - The problem of postoperative pain: issues for future research. AB - Uncontrolled postoperative pain continues despite abundant research in the area. The purposes of the paper are to review how past research influences our understanding of pain in the postsurgery context and to argue for a methodological shift towards naturalistic inquiry. Such a shift incorporates the complexities of pain assessment and management in the clinical practice environment. Decisions regarding pain are often examined outside of the contextual concerns of clinical practice. Research approaches have involved analyses of nurse and patient-related factors associated with pain. These approaches do not account for complex interactions that occur between nurses, patients and the dynamic environment in which these interactions take place. The failure of research to address the context of pain decisions has several implications. It limits our understanding of why pain continues despite ongoing research and it does not enable evaluation of clinical strategies to improve pain decision-making and pain outcomes for patients. PMID- 15544582 TI - Jordanian pregnant women's expectations of their first childbirth experience. AB - Limited numbers of studies have addressed Jordanian women's overall views and expectations of their first childbirth experience. A descriptive design using a convenience sample of 77 primigravid women was included in the analyses out of the total sample of 99 women. Participants responded to two valid and reliable questionnaires developed for the purpose of the study: the Expectations of Childbirth Experience questionnaire and the Expectations of Nursing Support During Labor and Birth questionnaire. The majority of primigravid women in this study expected a negative first childbirth experience. Findings indicated that the majority of women expected their overall childbirth experience to be frightening, very long, too difficult and painful. The expectations of a negative childbirth experience could be explained by limited labour preparation and advice given to women. Findings also indicated that women expected inadequate nursing and midwifery support during childbirth. Health-care providers should emphasize the importance of childbirth preparation and improving the quality of intranatal care to help in changing negative childbirth expectations and experiences of Jordanian women. PMID- 15544583 TI - A historical perspective on the contrasting experiences of nurses as research subjects and research activists. AB - According to the spectrum of opinion, nursing research appears to be alive but vulnerable, poised for further growth but still liable to wither if starved of funding. By using a historical approach to provide an understanding of the present, this paper hopes to offer some guidance for the healthy future of nursing research. Documentary and oral evidence has been cited to compare the experiences of nurses as the subjects of research and as researchers themselves. Investigations on recruitment and retention in nursing undertaken prior to World War II tended to give monocausal explanations for complex problems. As a result, pioneer nurse researchers had to work in an atmosphere of suspicion from sections of the profession and little regard from more established professions. The paper demonstrates that in order to make progress they used formal and informal support networks. Contemporary practitioners could identify similar networks. PMID- 15544584 TI - Positive outcomes for survivors of haematological malignancies from a spiritual perspective. AB - The findings indicate that there are many potentially positive outcomes, couched in terms of a spiritual journey, to be gained from the experience of serious illness for survivors who are well supported and obtain successful results from treatment. The work is from a qualitative programme exploring the notion of spirituality. The data are from the thematic analysis of verbatim transcriptions of audio-taped, in-depth, open-ended interviews with 12 survivors of haematological malignancies. The results indicate increased confidence and assertiveness, less dependence on the approval of others, greater ability to assert personal needs, increased awareness of body needs, being less judgmental and more compassionate. The positive outcomes also included the gift of extra life, the desire to live life to the fullest, a stronger sense of family togetherness, an awareness of reliable friends and family members, increased respect from others, changes in work values and an overall improvement in quality of life. PMID- 15544585 TI - The influence of the full moon on the number of admissions related to gastrointestinal bleeding. AB - The objective of this study was to analyse whether the number of admissions for gastrointestinal bleeding to our bleeding unit increases during the full moon. In a prospective study, we included 447 consecutive patients with gastrointestinal haemorrhage admitted to our bleeding unit during a period of two years. The number of admissions was allocated to the corresponding day of the lunar cycle, and full moon and non-full moon days were compared. A wide variation in the number of admissions throughout the lunar cycle was observed. There were 26 admissions on the 25 days of full moon and 421 admissions in the remaining 713 days of non-full moon. This difference was mainly related to a higher incidence of haemorrhage in men and variceal haemorrhage at full moon. The results of this study suggest an increase in the number of admissions related to gastrointestinal haemorrhage in our bleeding unit during the full moon, especially in men and in patients experiencing variceal haemorrhage. However, the wide variation in the number of admissions throughout the lunar cycle could limit interpretation of the results. Therefore, further studies are needed to clarify the possible influence of the moon on gastrointestinal haemorrhage. PMID- 15544586 TI - What impact will the immune tolerance network have on the future treatment of allergic diseases? AB - The immune tolerance network (ITN) plans to capitalize on the current state of understanding of immunologic science to solicit, design and implement human trails designed to 'cure' allergic diseases. While the ITN can bring tremendous resources to bear, it is actively looking for the participation of innovative physician-scientists worldwide in order to optimize the changes of success and thereby have a major impact on the future treatment of allergic diseases. PMID- 15544587 TI - Mast cell contribution to angiogenesis related to tumour progression. AB - The current wisdom is that tumours are endowed with an angiogenic capability and that their growth, invasion and metastasis are angiogenesis dependent. It is now well documented that neoplastic cells are influenced by their microenvironment and vice versa. The specific organ microenvironment determines the extent of cancer cell proliferation, angiogenesis, invasion and survival. Tumour cells are surrounded by an infiltrate of inflammatory cells, namely lymphocytes, neutrophils, macrophages and mast cells (MCs), which communicate via a complex network of intercellular signalling pathways, mediated by surface adhesion molecules, cytokines and their receptors. This review article summarizes: (i) the MC mediators involved in angiogenesis; (ii) the experimental evidence concerning the role played by MCs in angiogenesis; (iii) the list of solid and haematological tumours in which a close relationship between angiogenesis, tumour progression and MCs has been demonstrated; (iv) the circumstances in which MCs are a critical source of angiogenic factors in vivo, and in such cases, the signals that regulate their production and secretion that need to be determined as a prelude to the elaboration of new therapeutic strategies associated with MC presence and activation. PMID- 15544588 TI - Prophylactic management of children at risk for recurrent upper respiratory infections: the Preventia I Study. AB - BACKGROUND: Given the morbidity and mortality of asthma and the recent dramatic increase in its prevalence, pharmacologic prophylaxis of this disease in children at risk would represent a major medical advance. OBJECTIVES: The Preventia I Study was designed to evaluate the efficacy and long-term safety of loratadine in reducing the number of respiratory infections in children at 24 months. A secondary objective was to investigate the benefit of loratadine treatment in preventing the onset of respiratory exacerbations. METHODS: Preventia I was a randomized placebo-controlled study involving 22 countries worldwide. The children were 12-30 months of age at enrollment and had experienced at least five episodes of ENT infections, and no more than two episodes of wheezing during the previous 12 months. Phase I was a 12-month double-blind period during which the children were treated with loratadine 5 mg/day (2.5 mg/day for childrenC, located within a stimulatory protein-1 binding site in the COX-2 promoter region, has been shown to have significantly lower promoter activity in vitro compared with the wild-type and was associated with decreased plasma levels of C-reactive protein after coronary artery bypass surgery. We hypothesized that this polymorphism, which may result in decreased COX-2 transcription, could be associated with more severe asthma, and/or aspirin intolerant asthma (AIA). OBJECTIVE: To determine the association between the 765G>C COX-2 polymorphism and asthma, disease severity and AIA in a large, well phenotyped Australian population. METHODS: PCR and restriction fragment length polymorphism analysis was used to characterize the polymorphism in an Australian Caucasian population of patients with mild (n=322), moderate (n=254) or severe (n=88) asthma and in non-asthmatic control subjects (n=512), as well as in patients with AIA (n=58). Genotype and allele association analyses were performed using chi(2) tests. RESULTS: The polymorphic -765C allele was present in approximately 30% of asthmatic patients and non-asthmatic controls. There was no association between the -765G>C polymorphism and asthma (P=0.920), disease severity (P=0.840), atopy (P=0.655) or AIA (P=0.841) in this population. CONCLUSION: Although the -765G>C polymorphism may have lower promoter activity and result in decreased COX-2 expression, it is not associated with asthma, disease severity, AIA or atopy in this Australian population. PMID- 15544596 TI - Detection of tryptase-, chymase+ cells in human CD34 bone marrow progenitors. AB - BACKGROUND: Mast cells (MCs) arise from haematopoietic stem cells. We have recently reported that CD34(+) progenitors derived from human bone marrow (BM) develop into tryptase+, chymase+ MCs when cultured in the presence of recombinant human stem cell factor (rhSCF) and recombinant human IL-6 (rhIL-6). In an experiment for the expression of chymase during differentiation, chymase+ cells were detected in human BM, but tryptase+ cells were not found. OBJECTIVE: The purpose of this study was to show the appearance of chymase+ cells in CD34(+) cells with an origin different from MC differentiation. METHODS: CD34(+) cells from human BM were sorted with anti-CD117 monoclonal antibody (mAb), and cytospins of CD34(+), CD34(+)CD117(+), or CD34(+)CD117(-) were prepared. These cells were cultured with rhSCF+rhIL-6 for 12 weeks. Some of the cells were subjected to either histological stain with Wright-Giemsa or immunocytochemistry with anti-chymase mAb. Real-time RT-PCR was also performed to compare the transcriptional level of chymase from each cell preparation. RESULTS: Chymase was expressed in CD34(+) cells as well as human MCs by immunocytochemistry. Substantial CD34(+)CD117(-) cells, but not CD34(+)CD117(+) cells, were stained immunocytochemically with anti-chymase mAb. For 1 week culture with rhSCF+rhIL-6, no cells expressed chymase in any preparation. Real-time RT-PCR revealed positivity for chymase mRNA in CD34(+) cells, but it reduced at 1 week of culture, and increased as cells developed into MCs. Chymase mRNA in CD34(+)CD117(+) cells was negligible compared with that in CD34(+)CD117(-). Tryptase mRNA was below the detectable level in CD34(+) cells, and increased along with MC differentiation. After 12 weeks of culture, CD34(+)CD117(+) developed predominantly into MCs, whereas CD34(+)CD117(-) developed into monocytes/macrophages. CONCLUSION: Our findings suggested that chymase is present not only in MCs but also in CD34(+)CD117(-) BM progenitors, but that its origin is different from the MC lineage. PMID- 15544597 TI - Induction of mast cell interactions with blood vessel wall components by direct contact with intact T cells or T cell membranes in vitro. AB - BACKGROUND: Mast cells exert profound pleiotropic effects on immune cell reactions at inflammatory sites, where they are most likely influenced not only by the extracellular matrix (ECM) and inflammatory mediators but also by the proximity of activated T lymphocytes. We recently reported that activated T cells induce mast cell degranulation with the release of TNF-alpha, and that this activation pathway is mediated by lymphocyte function-associated antigen-1 (LFA 1)/intercellular adhesion molecule-1 (ICAM-1) binding. OBJECTIVE: To determine how this contact between the two cell types can modulate mast cell behaviour in an inflammatory milieu by examining the adhesion of mast cells to endothelial cells and ECM ligands in an integrin-dependent manner. METHODS: Human mast cells (HMC-1) were co-cultured with resting or activated T cells followed by testing their adhesion to endothelial cell and ECM ligands, stromal derived factor-1alpha (SDF-1alpha)-induced migration, and western blotting. RESULTS: Co-culturing HMC-1 with activated, but not with resting T cells resulted in marked stimulation of mast cell adhesion to vascular cell adhesion molecule-1 and ICAM-1 in a very late antigen-4- and LFA-1-dependent fashion. In addition, activated T cells or T cell membranes promoted HMC-1 adhesion to fibronectin (FN) and laminin. This effect was accompanied by the phosphorylation of extracellular regulated kinase and p38, but not of c-Jun N-terminal kinase. Importantly, the adhesive property of mast cells depended exclusively on the direct contact between the two cell types, since neither supernatants from activated T cells nor separation of the two cell populations with a porous membrane affected mast cell adhesion to FN. Furthermore, similar results were obtained when mast cells were incubated with purified membranes from activated T cells. These results suggest that, in addition to stimulating mast cell degranulation, the proximity of activated T lymphocytes to mast cells can mediate the adhesion of mast cell precursors to the endothelial ligands and ECM. Activated T cells also stimulated SDF-1alpha-induced mast cell migration. CONCLUSION: This symbiotic relationship between the two types of immune cells may serve to direct mast cells to specific sites of inflammation where their effector functions are required. PMID- 15544598 TI - Fel d 4, a cat lipocalin allergen. AB - BACKGROUND: Cat allergy is unique among allergy to mammals in that the major allergen Fel d 1 is a uteroglobin-like protein and not a lipocalin. The biochemical spectrum of the cat allergens is thus uncertain, particularly with regard to the role that a cat lipocalin protein may play in sensitization to cats in allergic individuals. OBJECTIVE: To analyse cDNA encoding a lipocalin allergen and the corresponding recombinant allergen at both the molecular and immunological levels. METHODS: A submandibular salivary gland cDNA expression library was constructed and screened for clones producing IgE-binding polypeptides. cDNA encoding a lipocalin allergen and its corresponding recombinant allergen were analysed. RESULTS: An IgE binding molecule with high sequence identity to the boar salivary lipocalin and the horse lipocalin Equ c 1 allergen was isolated and designated, Fel d 4. Serum from 62.96% of cat-allergic subjects examined had measurable IgE antibody to Fel d 4 but typically at low levels. Despite this in 47% of sera the anti-Fel d 4 IgE titres were higher than the anti-Fel d 1 titres. IgE binding to the lipocalin allergen could be blocked by an allergen extract from cow and to a lesser degree by extracts from horse and dog. CONCLUSION: Fel d 4 is a lipocalin allergen produced by the cat, which binds IgE at relatively high frequency in cat-sensitive individuals. The allergen provides not only a means for investigating differences in the immune response to lipocalin allergens from that found for other mammalian species but also an important reagent for the diagnosis of cat allergy. PMID- 15544599 TI - Characterization of cross-reactive bell pepper allergens involved in the latex fruit syndrome. AB - BACKGROUND: Between 30% and 50% of individuals who are allergic to latex products are also allergic to specific plant foods, a fact that is well documented as the latex-fruit syndrome. Simultaneous sensitization to latex and bell pepper has been previously reported. Although bell pepper fruits are frequently consumed raw, cooked or as a spice, little is known about the cross-reactive allergens. OBJECTIVE: In this study we wished to identify bell pepper allergens involved in the latex-fruit syndrome. METHODS: Sera of four patients who displayed clinical symptoms to latex and bell pepper were used in immunoblot studies on protein extracts of three different cultivars of fresh bell pepper and fresh Hevea latex. Cross-reactive allergens were identified by inhibition experiments using recombinant Hev b 8 (latex profilin), and natural Hev b 2 (latex beta-1,3 glucanase) in addition to the protein extracts. A novel cross-reactive IgE reactive 30 kDa protein was subjected to sequence analysis. RESULTS: Three patients displayed IgE to profilins from bell pepper fruits and latex. Two patients possessed IgE to Hev b 2, a latex beta-1,3-glucanase, and a homologous protein in bell pepper. One patient possessed IgE reactive with a protein of 30 kDa identified by N-terminal sequencing as an l-ascorbate peroxidase and another patient to a protein of 38 kDa. Additionally, IgE binding proteins in two higher molecular weight ranges showed cross-reactive capacities. CONCLUSION: Our findings show on the molecular level that bell pepper is part of the latex-fruit syndrome. For the first time we have identified the major latex allergen Hev b 2, a beta-1,3-glucanase, and the bell pepper l-ascorbate peroxidase as cross reactive allergens. We were also able to show that profilins are responsible for some of the IgE cross-reactivity. PMID- 15544600 TI - Vicilin and convicilin are potential major allergens from pea. AB - BACKGROUND: Allergic reactions to pea (Pisum sativum) ingestion are frequently associated with lentil allergy in the Spanish population. Vicilin have been described as a major lentil allergen. OBJECTIVE: To identify the main IgE binding components from pea seeds and to study their potential cross-reactivity with lentil vicilin. METHODS: A serum pool or individual sera from 18 patients with pea allergy were used to detect IgE binding proteins from pea seeds by immunodetection and immunoblot inhibition assays. Protein preparations enriched in pea vicilin were obtained by gel filtration chromatography followed by reverse phase high-performance liquid chromatography (HPLC). IgE binding components were identified by means of N-terminal amino acid sequencing. Complete cDNAs encoding pea vicilin were isolated by PCR, using primers based on the amino acid sequence of the reactive proteins. RESULTS: IgE immunodetection of crude pea extracts revealed that convicilin (63 kDa), as well as vicilin (44 kDa) and one of its proteolytic fragments (32 kDa), reacted with more than 50% of the individual sera tested. Additional proteolytic subunits of vicilin (36, 16 and 13 kDa) bound IgE from approximately 20% of the sera. The lentil vicilin allergen Len c 1 strongly inhibited the IgE binding to all components mentioned above. The characterization of cDNA clones encoding pea vicilin has allowed the deduction of its complete amino acid sequence (90% of sequence identity to Len c 1), as well as those of its reactive proteolytic processed subunits. CONCLUSIONS: Vicilin and convicilin are potential major allergens from pea seeds. Furthermore, proteolytic fragments from vicilin are also relevant IgE binding pea components. All these proteins cross-react with the major lentil allergen Len c 1. PMID- 15544601 TI - Cry j 1 isoforms derived from Cryptomeria japonica trees have different binding properties to monoclonal antibodies. AB - BACKGROUND: We identified five Cryptomeria japonica trees producing Cry j 1 isoforms that cannot be detected in a sandwich ELISA using two monoclonal antibodies, J1B01 and J1B07, suggesting that the binding affinity of these isoforms for both monoclonal antibodies is low. OBJECTIVES: The binding properties of the Cry j 1 isoforms produced by five trees to J1B07 and J1B01 were examined. The complementary DNA (cDNA) sequences of the Cry j 1 isoforms were also determined. METHODS: To clarify the binding properties of these Cry j 1 isoforms to J1B01 and J1B07, Cry j 1 was quantified in pollen samples collected from each of the five trees, by sandwich ELISAs using polyclonal antibodies and either J1B01 or J1B07. The cDNA sequences of isoforms with different binding properties were determined. To test the assumption that amino acid substitutions affect the binding affinities of Cry j 1 isoforms for monoclonal antibodies, cleaved amplified polymorphic sequences (CAPS) markers representing the putative polymorphisms were used to analyse additional trees. RESULTS: Four of the five trees produced Cry j 1 isoforms with extremely low binding affinity for J1B07, whereas the other tree produced two different isoforms with low binding affinity for either J1B01 or J1B07. Cry j 1-encoding cDNA sequences for one of the four trees and for the exceptional fifth tree indicate that amino acid substitutions at positions 55 and 352 in mature Cry j 1 affect its binding to J1B01 and J1B07, respectively. This was supported by the results of CAPS analysis. CONCLUSION: The existence of Cry j 1 isoforms with low binding affinity for either J1B01 or J1B07 was established. Furthermore, a single amino acid substitution is involved in this difference in binding affinity for each monoclonal antibody. PMID- 15544602 TI - Immunoglobulin E reactivity of native Blo t 5, a major allergen of Blomia tropicalis. AB - BACKGROUND: Blo t 5 is a major allergen of Blomia tropicalis and its complementary DNA (cDNA) has been expressed in both prokaryotic and eukaryotic expression systems. Although the recombinant Blo t 5 has been well characterized, relatively less is known about its native counterparts and the allergenicity comparison of the native and recombinant Blo t 5 allergens has not been reported. OBJECTIVE: The aims of this study are to characterize the native counterpart of Blo t 5, and compare the allergenicity of native and recombinant Blo t 5 by in vivo and in vitro assays. METHODS: Native Blo t 5 were purified by immuno affinity chromatography and characterized by proteomic means. The allergenicity of the allergen was evaluated by skin prick tests, human IgE ELISA, ELISA inhibition and histamine release assays. RESULTS: Native Blo t 5 consists of at least five distinct isoforms, ranging from pI 3 to 5.5. Allergenicity assessment of recombinant and native Blo t 5 based on skin reaction, IgE-binding capacity and histamine release in allergic individuals indicated that there was a good correlation between both forms of Blo t 5 in general. However, data from IgE ELISA inhibition assay revealed the presence of additional unique IgE epitopes in native Blo t 5. CONCLUSIONS: At least five distinct isoforms of Blo t 5 have been identified. Comparative assessment of native and recombinant Blo t 5 revealed that the allergenicity of these two forms was similar but not completely identical suggesting that the various isoforms of native Blo t 5 may exhibit additional unique IgE epitopes. PMID- 15544603 TI - The diagnostic interpretation of basophil activation test in immediate allergic reactions to betalactams. AB - BACKGROUND: Basophil activation by allergens, including drugs, has been used to determine sensitivity and to study IgE recognition and cross-reactivity. OBJECTIVE: We sought to determine the sensitivity and specificity of a basophil activation test (Basotest) in patients with immediate allergic reactions to betalactams, with a later comparison between patients who were selective (those recognizing the culprit drug excluding benzylpenicillin (BP)) and cross-reactors (those recognizing several penicillin determinants including BP). METHODS: Basotest to different haptens was performed in 70 patients with immediate allergic reactions to betalactams, classified into three groups: (A) skin test positive independently of CAP/RAST immunoassay value, (B) skin test negative and CAP/RAST positive, and (C) skin test and CAP/RAST negative but drug provocation test positive. Basotest was carried out by flow cytometry following the manufacturer's instructions using different betalactam determinants and results expressed as a stimulation index. RESULTS: Of the 70 patients, 34 (48.6%) were positive to Basotest (sensitivity: 48.6%), 31 (44.3%) to CAP/RAST and 46 (65.7%) to either one or the other. Considering the different groups, Basotest was positive in 50.9% of patients in Group A, 60% in Group B and 14.3% in Group C. The specificity was 91.3%. Positivity to the haptens was 28.6% to amoxicillin (AX), 21.7% to BP, 20% to benzylpenicilloyl-poly-l-lysine, 12.5% to ampicillin and 2.2% to minor determinant mixture. In patients with cephalosporin reactions, Basotest to the culprit cephalosporin was positive in 77.7%. There were differences between the two reactor groups in the sensitivity of Basotest (selective to AX=50%, cross-reactors=28.6%; chi(2)=10.809, P=0.004) and in the CAP/RAST (selective to AX=28.6%, cross-reactors=61.9%; chi(2)=8.944, P=0.011). CONCLUSIONS: The sensitivity of Basotest is similar to immunoassays (CAP/RAST). Sensitivity is improved when used in combination. Although further studies are required, Basotest results for cephalosporin allergy seem very promising. This technique does not help differentiate between selective reactors and cross reactors. PMID- 15544604 TI - Constant infusion of epinephrine, but not bolus treatment, improves haemodynamic recovery in anaphylactic shock in dogs. AB - OBJECTIVE: Epinephrine (Epi) is the treatment of choice for reversing cardiovascular collapse in anaphylactic shock (AS). In this condition, most treatment guidelines have been anecdotally derived and no randomized clinical trials have been conducted. In the present study, we examined the time course of haemodynamic recovery in a canine model of AS when Epi was administered at the initiation of allergen challenge before fully developed shock had occurred. METHODS: Randomized, controlled, crossover studies were performed approximately 3 5 weeks apart in ragweed-sensitized dogs while the animals were ventilated and anaesthetized. Epi was administered by bolus intravenous (i.v.), subcutaneous (s.c.), intramuscular (i.m.) routes and by continuous i.v. infusion (CI). The findings obtained in the Epi treatment (T) studies were compared with those found in a no treatment (NT) study. In the bolus studies, Epi was administered at 0.01 mg/kg, while in the CI study, the dose of Epi was titrated to maintain mean arterial pressure (MAP) at 70% of preshock levels. MAP, cardiac output (CO), stroke volume (SV), and pulmonary wedge pressure (Pwp) were determined over a 3 h period. RESULTS: In the CI study, haemodynamics (CO, MAP, and SV) were significantly higher than those measured in the NT study and the bolus studies over approximately the first hour of the study. In the CI study, the amount of Epi infused was significantly less than in the bolus studies. CONCLUSION: When administered at the initiation of allergen challenge, bolus treatment of Epi by i.m., i.v., or s.c. routes caused limited haemodynamic improvement in AS. In contrast, constant infusion of Epi at a lower total dose produced significant haemodynamic improvement. Within the limits of this anaesthetized canine model, the results suggest that CI should be the preferred route in the treatment of AS when this treatment option is available. PMID- 15544605 TI - Effects of lysed Enterococcus faecalis FK-23 on allergen-induced serum antibody responses and active cutaneous anaphylaxis in mice. AB - BACKGROUND: Our previous studies have presented evidence that lysed Enterococcus faecalis FK-23 (LFK), a lysozyme and heat-treated probiotic product, can inhibit allergen-induced local accumulation of eosinophils in mice. OBJECTIVE: The purpose of this experimental study was to evaluate the influence of orally administrated LFK on the host immune responses. METHODS: BALB/c mice were sensitized subcutaneously, and challenged intraperitoneally by cedar pollen allergen. Blood and spleen samples were collected after oral administration of LFK 60 mg/day for 21 days. The serum levels of total and allergen-specific IgE and IgG2a antibodies and the production of IL-4, IL-5 and IFN-gamma generated by allergen-stimulated cultured splenocytes were determined. Additionally, the effect of LFK on active cutaneous anaphylaxis (ACA) induced by ovalbumin (OVA) challenge in mice was measured after 28 days LFK treatment. RESULTS: No significant differences in serum immunoglobulin levels, as well as in cytokine production of splenocytes were observed between LFK-treated and control mice (P>0.05). There was, however, an increasing tendency of allergen-specific IgG2a level in mice after LFK treatment for 21 days compared with controls (P=0.060). Furthermore, the serum ratio of specific IgE to IgG2a was found to be significantly decreased in the LFK group (P=0.005). In addition, a significant inhibition of OVA-induced ACA reaction was observed in mice that had been fed for 28 days with LFK compared with control mice (P=0.008). CONCLUSION: These results suggest that LFK shows an anti-inflammatory effect, which may be part of the mechanism for protection against IgE-mediated allergy. PMID- 15544606 TI - Dose-dependent effects of endotoxins on allergen sensitization and challenge in the mouse. AB - BACKGROUND: Levels of endotoxins greatly differ according to environmental settings. OBJECTIVE: To study the effect of lipopolysaccharide (LPS) at increasing doses (0.1-1000 ng) on allergen sensitization and challenge in the mouse. METHODS: Mice were sensitized systemically and challenged locally with ovalbumin (OVA) in the presence or absence of LPS. Inflammation was assessed by determining total and differential cell counts and T-helper type 2 (Th)2 cytokine (IL-4 and IL-5) levels in bronchoalveolar lavage fluid (BALF). Total and OVA specific IgE levels were quantified in serum. Airway hyper-responsiveness (AHR) was assessed by whole-body barometric plethysmography. RESULTS: Administered prior to sensitization, LPS at 100 or 1000 ng dose-dependently decreased allergen induced total and OVA-specific IgE, airway eosinophilia and Th2 cytokines in BALF, without changing AHR. Administered during OVA challenge, LPS at 1 ng (an infra-clinical dose) or 100 ng (a dose triggering neutrophilia) enhanced airway eosinophilia, without affecting IgE levels or AHR. CONCLUSION: Our data clearly demonstrate that exposure to LPS influences allergen-induced IgE production and airway eosinophilia in a time and dose-dependent manner, preventing IgE production and development of eosinophilia when administered during allergen sensitization at high doses, and inducing exacerbation of eosinophilia when administered upon allergen challenge at low doses, including infra-clinical doses. PMID- 15544607 TI - Reversibility of airway inflammation and remodelling following cessation of antigenic challenge in a model of chronic asthma. AB - BACKGROUND: Asthma is associated with recruitment of eosinophils, accumulation of chronic inflammatory cells in the airway walls, subepithelial fibrosis and other structural changes of airway wall remodelling. The role of ongoing exposure to allergens in their pathogenesis remains unclear. OBJECTIVE: To examine whether changes of inflammation and remodelling were reversible following cessation of antigenic challenge in a mouse model of chronic asthma. METHODS: BALB/c mice sensitized to ovalbumin (OVA) were chronically challenged by inhalation of a low mass concentration of antigen for 8 weeks, leading to development of acute-on chronic airway inflammation, subepithelial fibrosis and other changes of airway wall remodelling. Epithelial injury was assessed by immunohistochemistry, while inflammation and remodelling were quantified by appropriate histomorphometric techniques. Regression of lesions was assessed in animals examined at 1, 2 and 4 weeks after exposure to OVA ceased. RESULTS: We did not find evidence of airway epithelial injury in this model of low-level chronic inhalational exposure to antigen. Persistence of the recruitment of eosinophils and chronic inflammatory cells in the airway walls was dependent on continuing antigenic challenge, as was persistence of mucous cell hyperplasia/metaplasia. Subepithelial fibrosis and epithelial hypertrophy exhibited delayed reversibility following cessation of exposure to antigen, possibly related to matrix-associated accumulation of transforming growth factor-beta(1). CONCLUSION: In chronic asthma, low-level antigenic challenge may be required to maintain the inflammatory response in the airway wall, but airway remodelling may persist in its absence. PMID- 15544608 TI - Comments regarding 'Assessment of histamine-releasing activity of sera from patients with chronic urticaria showing positive autologous skin test on human basophils and mast cells' by Asero et al. M. Ferrer. PMID- 15544609 TI - Comments regarding 'Assessment of histamine-releasing activity of sera from patients with chronic urticaria showing positive autologous skin test on human basophils and mast cells' by Asero et al. A.P. Kaplan. PMID- 15544612 TI - Early up-regulation of Th2 cytokines and late surge of Th1 cytokines in an atopic dermatitis model. AB - We investigated cytokine profiles in interleukin (IL)-4 transgenic (Tg) mice with a skin inflammatory disease resembling human atopic dermatitis. cDNA microarray revealed that the mRNAs encoding IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, IL-12p40, IL-13, tumour necrosis factor (TNF)-alpha, TNF-beta and interferon (IFN)-gamma were up-regulated in the skin of late lesion Tg mice and to a lesser degree in non-lesion Tg mice when compared to those of non-Tg mice. Real time reverse transcription-polymerase chain reaction (RT-PCR) analyses indicated that the cDNA copy numbers of IL-1beta, IL-4, IL-6, IL-10, TNF-alpha and IFN-gamma from the skin of late, early and non-lesions increased significantly compared to non-Tg mice. IL-2 and IL-12p40 cDNA copy numbers were increased significantly in early, but not late, lesions. Interestingly, IL-1beta, IL-3, IL-4, IL-5, IL-6, IL 10, IL-13, TNF-alpha, and IFN-gamma cDNAs were increased significantly the skin of before-onset and/or non-lesion mice. Flow cytometry analyses demonstrated an increased percentage of keratinocytes producing IL-4 as the disease progressed. The percentage of IL-2, IL-4, IL-10 and IFN-gamma-producing T cells and IL-12 producing antigen-presenting cells in skin-draining lymph nodes and inflammatory skin also increased, particularly in mice with late lesion. These results suggest that disease induction is primarily triggered by Th2 cytokines and that Th1, Th2 and non-Th proinflammatory cytokines are all involved in the disease process. PMID- 15544613 TI - Bovine respiratory syncytial virus infection influences the impact of alpha- and beta-integrin-mediated adhesion of peripheral blood neutrophils. AB - Neutrophil migration into the airways and pulmonary tissue is a common finding in bovine respiratory syncytial virus (BRSV) infections. Although neutrophil trans endothelial migration in general depends on beta2-integrins, alternative integrins such as the alpha4-integrins have been implicated. In this study, rolling and firm adhesion of peripheral blood neutrophils isolated from healthy and BRSV-infected calves to tumour necrosis factor (TNF)-alpha activated pulmonary endothelium was investigated under flow conditions in vitro. For neutrophils obtained from healthy animals, inhibition of the beta2-integrin reduced firm adhesion to 63% and inhibition of alpha4-integrin to 73% compared with untreated controls. Inhibition of both integrins reduced firm adhesion to 25%. Rolling velocity, which is used as a parameter for integrin involvement in neutrophil rolling, increased 1.7-fold by blocking beta2-integrin and was significantly augmented to 2.5-fold by blocking both alpha4- and beta2-integrins. For neutrophils obtained from BRSV-infected animals, however, rolling velocities at 10 days after infection (p.i.) were not influenced by blocking adhesion of alpha4- and beta2-integrins, indicating that these integrins did not support neutrophil rolling. In addition, the inhibition of firm adhesion by blocking both alpha4- and beta2-integrins was reduced significantly 9 days post-infection, resulting in a residual 68% neutrophil binding at 9 days p.i. Non-blocked firm adherence was not reduced, indicating that binding was achieved by other mechanisms than through alpha4- and beta2-integrins. These results demonstrate an important function for alpha4- and beta2-integrins in rolling and firm adherence of bovine neutrophils, to TNF-alpha-activated endothelium and show the dynamic use of these integrins for adhesion and migration by neutrophils in the course of BRSV infection. PMID- 15544614 TI - Co-infection of malaria and gamma-herpesvirus: exacerbated lung inflammation or cross-protection depends on the stage of viral infection. AB - In order to study the interaction between a gamma-herpesvirus and malaria we established a co-infection model that involves infection of mice with murine gamma-herpesvirus (MHV-68) and Plasmodium yoelii non-lethal strain (PYNL). To investigate the interaction between acute malaria and the lytic stage of MHV-68, the timing of infections was chosen such that the peak virus and parasite burdens would be present at the same time. Under this condition, we observed significant mortality in co-infected mice and aggressive lung inflammation with a marked influx of neutrophils and megakaryocytes. If mice were latently infected with MHV 68 and then co-infected with malaria we noticed significantly less viral load and parasitaemia. Using MHC/peptide tetramer staining we found that acute malaria reduces the anti-MHV-68 CD8+ T cell response in the animals that develop severe disease. Our study provides important fundamental information, which will be of use when devising strategies to combat infections with more than one agent, a situation that often occurs naturally. PMID- 15544615 TI - Intranasal bacille Calmette-Guerin (BCG) vaccine dosage needs balancing between protection and lung pathology. AB - Intranasal vaccination may offer practical benefits and better protection against respiratory infections, including tuberculosis. In this paper, we investigated the persistence of the Mycobacterium bovis-strain bacille Calmette-Guerin (BCG) Pasteur, lung granuloma formation and protection against pathogenic tuberculous challenge in mice. A pronounced BCG dose-dependent granulomatous infiltration of the lungs was observed following intranasal, but not after subcutaneous, vaccination. Corresponding doses of BCG, over a 100-fold range, imparted similar protection against H37Rv challenge when comparing the intranasal and subcutaneous vaccination routes. Interestingly, a BCG dose-dependent reduction of the H37Rv challenge infection was observed in the lungs, but not in the spleens, following both intranasal and subcutaneous vaccination. In the light of the observed concurrence between the extent of granuloma formation and the level of protection of the lungs, we conclude that intranasal vaccination leading to best protective efficacy needs to be balanced with an acceptable safety margin avoiding undue pathology in the lungs. PMID- 15544616 TI - Low electric field enhanced chemotherapy can cure mice with CT-26 colon carcinoma and induce anti-tumour immunity. AB - Low electric field cancer treatment-enhanced chemotherapy (LEFCT-EC) is a new anticancer treatment which utilizes a combination of chemotherapeutic agents and a low electric field. We investigated the antitumour effectiveness of this technique in a model of murine colon carcinoma (CT-26). The low electric field was applied to approximately 65 mm3 intracutaneous tumours after intratumoral injection of 5FU, bleomycin or BCNU. We observed significant tumour size reduction and a prolongation of survival time. The complete cure of a significant fraction of animals treated by LEFCT-EC with 5FU (33%), bleomycin (51%) or BCNU (83%) was observed. Mice cured by LEFCT-EC developed resistance to a tumour challenge and their splenocytes had antitumour activity in vivo. Our results suggest that LEFCT-EC is an effective method for treatment of solid tumours. PMID- 15544617 TI - TSH receptor-adenovirus-induced Graves' hyperthyroidism is attenuated in both interferon-gamma and interleukin-4 knockout mice; implications for the Th1/Th2 paradigm. AB - The role of the Th1/Th2 balance in the pathogenesis of murine Graves' hyperthyroidism is controversial. In BALB/c mice injected with adenovirus expressing TSH receptor (TSHR-adeno model), we found that suppression of TSHR specific Th1 immune responses by exogenous interleukin-4 (IL-4), alpha galactosylceramide or helminth (Schistosoma mansoni) infection was associated with inhibition of hyperthyroidism, indicating the critical role for Th1 cytokines. In contrast, BALB/c IL-4 knockout (KO), but not interferon-gamma (IFN gamma) KO mice failed to develop Graves' hyperthyroidism when injected with TSHR expressing M12 B lymphoma cells (TSHR-M12 model), suggesting the importance of Th2 cytokine IL-4. To reconcile differences in these two models, we used IL-4 KO and IFN-gamma KO BALB/c mice in the TSHR-adeno model. Unlike wild-type (wt) BALB/c mice in which 60% developed hyperthyroidism, only 13 and 7% of IL-4 KO and IFN-gamma KO mice, respectively, became hyperthyroid. Thyroid stimulating antibodies were positive in most hyperthyroid mice. TSHR antibody titres determined by TSH binding inhibition and ELISA were comparable in all three groups. IgG1 and IgG2a TSHR antibody titres were similar in IFN-gamma KO and wt mice, whereas IgG1 TSHR antibody titres and TSHR-specific splenocyte IFN-gamma secretion were lower in IL-4 KO than in IFN-gamma KO and wt mice, respectively. Our results clearly implicate both IFN-gamma and IL-4 in development of hyperthyroidism in the TSHR-adeno model. These data, together with the previous report, also indicate different cytokine requirements in these two Graves' models, with IFN-gamma being more important in the TSHR-adeno than the TSHR-M12 model. Moreover, our previous and present observations indicate a difference in the role of exogenous versus endogenous IL-4 in TSHR-adenovirus induced Graves' hyperthyroidism. PMID- 15544618 TI - Antibodies cross-reacting with thyroglobulin and thyroid peroxidase are induced by immunization of rabbits with an immunogenic thyroglobulin 20mer peptide. AB - Thyroglobulin (Tg) and thyroid peroxidase (TPO) are two major autoantigens in autoimmune thyroid diseases (AITD). Cross-reactive anti-Tg/TPO antibodies have been identified in patients with AITD and in mice immunized with Tg or TPO. In the present study, we investigated the production of anti-Tg/TPO antibodies in rabbits immunized with human Tg and with a highly immunogenic Tg peptide (namely TgP41, sequence 2651-2670 of human Tg), by noncompetitive and competitive ELISA. TgP41 was found previously to induce intramolecular epitope spreading. We found that Tg-immunized rabbits developed a serological immune response to TPO due to cross-reactivity with Tg, since serum TPO reactivity was inhibited by soluble Tg and affinity-purified anti-Tg antibodies cross-reacted with TPO. Moreover, TgP41 immunized rabbits responded to Tg and TPO. This serological response was attributed to anti-Tg/TPO antibodies, based on the observation that serum TPO reactivity was again inhibited by soluble Tg, and affinity-purified anti-Tg antibodies, induced by TgP41-immunization, cross-reacted with TPO. Purified anti TgP41 antibodies did not react with TPO, suggesting that a putative common antigenic determinant is not included in the peptide sequence. We propose that intermolecular spreading of reactivity to TPO observed after administration of the Tg-peptide is a result of intramolecular epitope spreading to determinant(s) responsible for Tg/TPO cross-reactivity. PMID- 15544619 TI - Blockade of the C5a receptor fails to protect against experimental autoimmune encephalomyelitis in rats. AB - Complement activation contributes to inflammation and tissue damage in human demyelinating diseases and in rodent models of demyelination. Inhibitors of complement activation ameliorate disease in the rat model antibody-dependent experimental autoimmune encephalomyelitis and rats unable to generate the membrane attack complex of complement develop inflammation without demyelination. The role of the highly active chemotactic and anaphylactic complement-derived peptide C5a in driving inflammation and pathology in rodent models of demyelination has been little explored. Here we have used a small molecule C5a receptor antagonist, AcF-[OPdChaWR], to examine the effects of C5a receptor blockade in rat models of brain inflammation and demyelination. C5a receptor antagonist therapy completely blocked neutrophil response to C5a in vivo but had no effect on clinical disease or resultant pathology in either inflammatory or demyelinating rat models. We conclude that C5a is not required for disease induction or perpetuation in these strongly complement-dependent disease models. PMID- 15544620 TI - The quantitative role of alternative pathway amplification in classical pathway induced terminal complement activation. AB - Complement activation with formation of biologically potent mediators like C5a and the terminal C5b-9 complex (TCC) contributes essentially to development of inflammation and tissue damage in a number of autoimmune and inflammatory conditions. A particular role for complement in the ischaemia/reperfusion injury of the heart, skeletal muscle, central nervous system, intestine and kidney has been suggested from animal studies. Previous experiments in C3 and C4 knockout mice suggested an important role of the classical or lectin pathway in initiation of complement activation during intestinal ischaemia/reperfusion injury while later use of factor D knockout mice showed the alternative pathway to be critically involved. We hypothesized that alternative pathway amplification might play a more critical role in classical pathway-induced C5 activation than previously recognized and used pathway-selective inhibitory mAbs to further elucidate the role of the alternative pathway. Here we demonstrate that selective blockade of the alternative pathway by neutralizing factor D in human serum diluted 1 : 2 with mAb 166-32 inhibited more than 80% of C5a and TCC formation induced by solid phase IgM and solid- and fluid-phase human aggregated IgG via the classical pathway. The findings emphasize the influence of alternative pathway amplification on the effect of initial classical pathway activation and the therapeutic potential of inhibiting the alternative pathway in clinical conditions with excessive and uncontrolled complement activation. PMID- 15544621 TI - Divalent cation-dependent and -independent augmentation of macrophage phagocytosis of apoptotic neutrophils by CD44 antibody. AB - Phagocytosis of apoptotic neutrophils by macrophages is required for resolution of an inflammatory response. Removal of intact apoptotic neutrophils prevents the release of cytotoxic granules that would otherwise cause tissue damage and may lead to development of fibrosis. Importantly, macrophage phagocytosis of apoptotic neutrophils fails to induce release of proinflammatory mediators, consistent with a 'safe' pathway for disposal of potentially harmful inflammatory cells. One pathway for increasing phagocytosis of apoptotic cells to allow matching of tissue phagocyte capacity to apoptotic cell load in vitro is via antibody-mediated cross-linking of CD44, providing a mechanism for limiting tissue damage during resolution of inflammation. In this study, we have defined divalent cation-dependent and -independent actions of the CD44 antibody. For the divalent cation-independent CD44 antibody effect, we provide evidence that 'enabled' CD32 on the apoptotic neutrophil binds to intact CD44 antibody on the macrophage surface. One implication is that macrophages can phagocytose apoptotic neutrophils that are 'tethered' to the macrophage surface in a manner that is independent of defined apoptotic mechanisms. These data also provide an explanation for the greater efficacy of intact CD44 antibody when compared with F(ab')2 fragments. PMID- 15544622 TI - Cl- channels are expressed in human normal monocytes: a functional role in migration, adhesion and volume change. AB - Increased adhesion and diapedesis of monocytes appear to be primary initiating factors in the pathophysiology of occlusive vascular diseases, including atherosclerosis and restenosis. However, the underlying mechanisms of transendothelial migration and invasion of monocytes into the blood vessels are not known. Alterations in ion channels on the cell membrane are generally involved in induced changes in shape and volume. In the present study, we investigated the expression and functional role of chloride channels in freshly isolated human blood monocytes. The Cl- currents in whole-cells were measured by the patch-clamp technique. We observed whole cell Cl- currents, which were time independent and outwardly rectifying. The chloride channel blockers 5-nitro-2-(3 phenylpropylamino) benzoic acid (NPPB) and R(+)-[(6,7-dichloro-2-cyclopentyl-2,3 dihydro-2-methyl-1-oxo-1H-inden-5yl)-oxy]acetic acid 94 (IAA94) attenuated the Cl currents. NPPB and IAA94 also inhibited chemotaxis of monocytes, as measured in Boyden chemotactic chambers, with the same sensitivity. NPPB but not IAA94, increased the cell volume as measured by shape change, and decreased tumour necrosis factor (TNF)-alpha-induced monocyte adhesion to endothelial cells. These results suggest that monocytes contain Cl- channels which regulate transendothelial migration of monocytes, due presumably to an alteration in cell volume. PMID- 15544623 TI - Differential expression of the cytokine receptors for human interleukin (IL)-12 and IL-18 on lymphocytes of both CD45RA and CD45RO phenotype from tonsils, cord and adult peripheral blood. AB - The objective of this study was to demonstrate the variable expression of cytokine receptors on naive versus memory human CD4+ T cell subpopulations in tonsillar tissue, cord blood and adult blood. We prove that the receptors for both interleukin (IL)-12 and IL-18 are expressed exclusively on memory T cells. This observation was seen not only on the CD45RO+ memory T cells but also on a significant percentage of the CD45RA+, CD62L-, CD27- and CCR7- populations. Furthermore, CD45RA+ CD62L+, CD27+ or CCR7+ CD4+ T cells that expressed IL 12Rbeta1 and IL-18Ralpha did not express CD31, a marker for recent thymic emigrants. We reveal that cord blood lymphocytes do not express IL-12Rbeta1 whereas IL-18Ralpha expression was detected at low levels. Importantly, the IL 12Rbeta2 signalling chain, which is absent in all resting T cells, was up regulated in both CD45RA+ and CD45RO+ T cells as a result of stimulation with anti-CD3 and anti-CD28 in vitro. This observed up-regulation was, however, restricted to 80% of the total CD4+ population. Finally, a very small proportion of the CD4+ CD45RO+ tonsillar T cells expressed the IL-12 and IL-18 receptors, thereby establishing the differential expression of these receptors between peripheral and tonsillar memory T cell subpopulations. PMID- 15544624 TI - Differential expression of receptors for advanced glycation end-products in peritoneal mesothelial cells exposed to glucose degradation products. AB - Autoclaving peritoneal dialysate fluid (PDF) degrades glucose into glucose degradation products (GDPs) that impair peritoneal mesothelial cell functions. While glycation processes leading to formation of advanced glycation end-products (AGE) were viewed commonly as being mediated by glucose present in the PDF, recent evidence indicates that certain GDPs are even more powerful inducers of AGE formation than glucose per se. In the present study, we examined the expression and modulation of AGE receptors on human peritoneal mesothelial cells (HPMC) cultured with GDPs, conventional PDF or PDF with low GDP content. HPMC cultured with GDPs differentially modulated AGE receptors (including RAGE, AGE R1, AGE-R2 and AGE-R3) expression in a dose-dependent manner. At subtoxic concentrations, GDPs increased RAGE mRNA expression in HPMC. 2-furaldehyde (FurA), methylglyoxal (M-Glx) and 3,4-dideoxy-glucosone-3-Ene (3,4-DGE) increased the expression of AGE-R1 and RAGE, the receptors that are associated with toxic effects. These three GDPs up-regulated the AGE synthesis by cultured HPMC. In parallel, these GDPs also increased the expression of vascular endothelial growth factor (VEGF) in HPMC. PDF with lower GDP content exerted less cytotoxic effect than traditional heat-sterilized PDF. Both PDF preparations up-regulated the protein expression of RAGE and VEGF. However, the up-regulation of VEGF in HPMC following 24-h culture with conventional PDF was higher than values from HPMC cultured with PDF containing low GDP. We have demonstrated, for the first time, that in addition to RAGE, other AGE receptors including AGE-R1, AGE-R2 and AGE-R3 are expressed on HPMC. Different GDPs exert differential regulation on the expression of these receptors on HPMC. The interactions between GDPs and AGE receptors may bear biological relevance to the intraperitoneal homeostasis and membrane integrity. PMID- 15544625 TI - The anti-CD2 monoclonal antibody BTI-322 generates unresponsiveness by activation associated T cell depletion. AB - The antihuman CD2 MoAb BTI-322 (Lo-CD2a) effectively inhibits T cell responses in vitro to allogeneic cells, which is followed by unresponsiveness to the original stimulator in secondary stimulation. We studied the xenogeneic human antiporcine mixed lymphocyte reaction (MLR), and utilized anti-T cell receptor (TCR) Vbeta family antibody-induced cell proliferation to determine the specificity and mechanism. BTI-322 and its humanized version, MEDI-507, effectively inhibited the primary xenogeneic MLR. After suboptimal primary stimulation using lower numbers of xenogeneic stimulator cells, the unresponsiveness in secondary culture was apparent only for xenogeneic stimulator cells of the original SLA haplotype, and not for third-party stimulators or allogeneic cells. The inhibition of primary MLR was not observed for nylon-wool-purified T cells, but was seen after reconstitution of purified T cells with monocytes. Similarly, anti-Vbeta family specific stimulation showed family-specific unresponsiveness in secondary culture. This required the presence of the whole BTI-322 molecule: a F(ab')2 fragment was not effective. T cells of a distinct Vbeta family were depleted after stimulation with an anti-Vbeta family-specific antibody and BTI-322. We conclude that the inhibition by BTI-322 of a primary xenogeneic MLR or the response to an anti-TCR Vbeta antibody is associated with unresponsiveness upon restimulation, due to activation-associated cell depletion. In this process, the interaction between monocytes and the Fc part of the antibody is involved. This unique characteristic of BTI-322 suggests the potential of the antibody for tolerance induction in vivo, besides the potential use as a T cell depleting agent. PMID- 15544626 TI - Monocyte derived dendritic cell responses in common variable immunodeficiency. AB - The phenotype and function of monocyte derived dendritic cells (MdDC) were investigated in 25 patients with common variable immunodeficiency (CVID) to test for abnormalities that might help explain the failure of antibody production. Using MHC class II DR and CD86 as markers of maturation, DCs from the majority of CVID patients were normal. However 5 patients, the majority of whom had affected family members who had previously been shown to have a susceptibility genetic locus in the MHC region, expressed abnormally low levels of DR on repeated testing, in some cases associated with a reduced capacity to support antigen stimulated T cell proliferation; nevertheless costimulatory molecules for production of IL-13, IL-10 and IFN-gamma from T cells were intact. In contrast to DCs from healthy donors, DCs from many CVID patients had high spontaneous production of IL-8 and lipopolysaccharide stimulation often caused a reduction in DR expression. Expression of other cytokines (IL-1a, IL-6 and IL-12), either before or after LPS stimulation, was normal. The data suggests there is a fundamental defect in the maturation of MdDCs in a subset of CVID patients that may compromise antigen presentation and subsequent antibody production. PMID- 15544627 TI - Increased expression of receptor activator of NF-kappaB ligand (RANKL), its receptor RANK and its decoy receptor osteoprotegerin in the colon of Crohn's disease patients. AB - Crohn's disease (CD) is associated with low bone mass due to chronic inflammation and other factors. Receptor activator of NF-kappaB ligand (RANKL), its receptor RANK and its decoy receptor osteoprotegerin (OPG) are potentially involved in this process as they regulate osteoclastogenesis and are influenced by pro inflammatory cytokines. The aim of this study was to determine the levels of soluble RANKL (sRANKL), RANK and OPG expression both in the serum and in the colon of CD patients. Levels of sRANKL and OPG were assessed in the serum and the supernatants of cultured colonic biopsies in patients with CD and controls by ELISA. RANK expression was explored by immunostaining and immunofluorescence of fixed colonic samples. OPG and sRANKL levels were higher in the serum of CD patients as compared to age- and sex-matched controls. Levels of sRANKL and OPG were significantly enhanced in cultured colonic biopsies from CD, and OPG levels correlated with histological inflammation, and pro- and anti-inflammatory cytokine levels. No significant correlation was found for sRANKL. RANK+ cells were increased in the colon of CD, particularly in inflamed areas. These cells were positive for CD68 or S100 protein. We conclude that serum and local levels of sRANKL and OPG are increased in CD. Moreover, RANK is expressed in the colonic mucosa by subpopulations of activated macrophages or dendritic cells at higher levels in CD compared to normal colon. PMID- 15544628 TI - Interferon-beta up-regulates the expression of co-stimulatory molecules CD80, CD86 and CD40 on monocytes: significance for treatment of multiple sclerosis. AB - Interferon (IFN)-beta reduces the biological activity of multiple sclerosis (MS), a presumably T cell-mediated autoimmune disease of central nervous system (CNS) myelin. Co-stimulatory molecules are necessary for full T cell activation and differential expression of co-stimulatory molecules on antigen-presenting cells is thought to influence the type of effector T cell response (Th1/Th2). In this study we investigated the effects of IFN-beta on the expression of co-stimulatory molecules on lymphocytes and monocytes as a potential mechanism of action of IFN beta in MS. Peripheral blood mononuclear cells (PBMCs) were stimulated with IFN beta in vitro and expression of CD80, CD86, CD40 and HLA was examined by flow cytometry and reverse-transcription polymerase chain reaction. Whereas IFN-beta had no effect on the expression of these molecules on T and B lymphocytes there was a significant increase on monocytes. Correspondingly, the expression of mRNA increased after 6-18 h. This in vitro response was also observed in untreated MS patients and patients receiving treatment with IFN-beta. The increase of co stimulatory molecules on monocytes was not mediated by interleukin (IL)-10. When IFN-beta-stimulated monocytes were used to stimulate autologous T cells an increased secretion of IL-13 was observed. In biopsies taken from IFN-beta induced skin reactions after subcutaneous injection increased expression of CD80 mRNA was detected, indicating that IFN-beta also up-regulates this co-stimulatory molecule in vivo. These data provide the background for further studies of IFN beta-induced changes of co-stimulatory molecules in MS patients. PMID- 15544629 TI - Characterization of a population of small macrophages in induced sputum of patients with chronic obstructive pulmonary disease and healthy volunteers. AB - The inflammatory process in chronic obstructive pulmonary disease (COPD) is active mainly in the airways, but little is known about the properties of the inflammatory cells in this compartment. We have studied leucocytes in induced sputum of COPD patients compared to controls in order to uncover what types of macrophages might be involved in the disease. Sputum induction was performed by inhalation of nebulized sodium chloride solution. Leucocytes were isolated and stained with specific monoclonal antibodies for analysis in flow cytometry. Flow cytometry analysis revealed that a major portion of CD14+ macrophages in COPD has lower forward scatter, i.e. they are small macrophages. While in control donors these small macrophages accounted for 6.9% of all macrophages, the percentage of these cells in COPD was 45.7%. CD14 and HLA-DR expression was high on these small sputum macrophages while the large sputum macrophages expressed only low levels of these surface molecules, both in control donors and COPD patients. Small sputum macrophages of both control donors and COPD patients showed higher levels of constitutive tumour necrosis factor (TNF) compared to the large macrophages. TNF was inducible by lipopolysaccharide (LPS) preferentially in the small sputum macrophages in the control donors but there was no further induction in COPD patients. These data show that the small sputum macrophages are a major macrophage population in COPD and that these cells exhibit features of highly active inflammatory cells and may therefore be instrumental in airway inflammation in COPD. PMID- 15544630 TI - L-ficolin in children with recurrent respiratory infections. AB - The lectin pathway of complement activation is used by a collectin, mannan binding lectin (MBL), and two ficolins, L-ficolin and H-ficolin, to opsonize microorganisms for phagocytosis. We published evidence recently that MBL insufficiency is associated with recurrent respiratory infections in childhood. We have now measured serum L-ficolin in 313 respiratory infection patients and 74 healthy control children. L-ficolin concentrations below the lower limit of the control group were found in 6% of the patients (P <0.02) and were associated most strongly with children having co-existing atopic disorders (11%; P=0.002). We suggest that L-ficolin may have a role in protection from microorganisms complicating allergic disease. PMID- 15544631 TI - Significantly increased levels of mannose-binding lectin (MBL) in rheumatic heart disease: a beneficial role for MBL deficiency. AB - Although mannose-binding lectin (MBL) is known to be involved in the primary defense against microorganisms, there are emerging lines of evidence for an active proinflammatory role for MBL in different chronic diseases. In this study we determined the circulating levels of MBL in patients with rheumatic heart disease (RHD). A total of 100 patients (77 women, 23 men; mean age 45.8 +/- 11 years, range 19-76 years) with chronic RHD, and a previous diagnosis of rheumatic fever, were studied. Transthoracic echocardiography was performed in all patients to evaluate valvular heart disease. Ninety-nine healthy individuals matched for age, sex and ethnic origin were included as controls. MBL concentration was measured by enzyme-linked immunosorbent assay and C3 and C4 levels by turbidimetry. MBL levels were significantly higher in patients with RHD than in healthy subjects (mean +/- SEM: 3036.2 +/- 298.9 ng/ml versus 1942.6 +/- 185.5 ng/ml, P <0.003). In addition, MBL deficiency was more prevalent in controls (17.1%) than in patients (9% P <0.09). Concentrations of C4 were within the normal range (22.7 +/- 0.8 mg/dl, normal: 10.0-40.0 mg/dl), while C3 concentrations were found to be elevated (109.2 +/- 3.6 mg/dl, normal: 50.0-90.0 mg/dl). No correlation was observed between serum MBL levels and valve area or the type of surgical procedure. The significantly elevated circulating MBL levels in patients with RHD together with the greater prevalence of MBL deficiency in controls suggest that MBL may cause undesirable complement activation contributing to the pathogenesis of RHD. PMID- 15544632 TI - Does immune activation continue during an attack-free period in familial Mediterranean fever? AB - Although some information is available regarding immune activation in familial Mediterranean fever (FMF), little is known about either peripheral blood T cell activation marker expression or the T cell proliferative response to phytohaemagglutinin (PHA). In the present study, we aimed to investigate the percentages of peripheral blood lymphocyte subsets, T cell expression of cellular activation markers (CD25, CD69, HLA-DR), the T cell response to PHA and serum levels of soluble interleukin-2 receptor (sIL-2R) and interleukin (IL)-10 in patients with FMF. Forty patients with FMF were enrolled into the study. Control groups were sex- and age-matched and consisted of 20 healthy blood donors and 15 patients with inactive Behcet's disease. The patients with FMF in an attack period had higher levels of sIL-2R than those in an attack-free period, and also in comparison with both control groups. The levels of sIL-2R were also found to be higher in patients with FMF in an attack-free period than those in both control groups. The mean levels of IL-10 were found to be lower in patients with FMF in an attack-free period than those in an attack period and were also lower than those in the healthy controls. In an acute attack period, the absolute counts of CD3+HLA-DR+, CD4+CD69+, CD8+CD25+ and CD8+CD69+ T cells in peripheral blood samples were also higher than those in both control groups. Both the percentages and absolute counts of CD4+CD69+ T cells in peripheral blood samples of patients with FMF in an attack-free period were slightly but significantly higher than those in the healthy controls. In conclusion, our study indicates that the T cell system is abnormally activated in patients with FMF in both the attack and attack-free period and that decreased IL-10 levels may create a tendency to perpetuate subclinical immune activation in the attack-free period. PMID- 15544633 TI - Autoantibodies to a collagen-specific molecular chaperone, heat-shock protein 47, in systemic sclerosis. AB - Heat-shock proteins are highly conserved and immunogenic proteins, which may be involved in the initiation and perpetuation of autoimmune diseases. Heat-shock protein 47 (HSP47) is expressed by collagen-secreting cells such as fibroblasts and serves as a collagen-specific molecular chaperone that plays a crucial role in collagen metabolism. Abnormal collagen accumulation and autoimmunity are characteristics of systemic sclerosis (SSc). We determined the presence and prevalence of autoantibodies to HSP47 in patients with SSc and also in tight-skin (TSK/+) mice, an animal model for SSc. Anti-HSP47 autoantibodies were present in SSc patients with a frequency of 26%, while patients with systemic lupus erythematosus, those with dermatomyositis, those with keloid and healthy subjects did not have anti-HSP47 antibodies. IgG1 and IgG2 were the major Ig isotypes of the autoantibodies. Patients positive for anti-HSP47 had a significantly shorter duration of disease than those who were negative. Anti-HSP47 autoantibodies were also positive in 79% of TSK/+ mice. Thus, autoantibodies to HSP47 were present in the sera from SSc patients as well as those from TSK mice, and may be associated with the pathogenesis of SSc. PMID- 15544635 TI - Cellular and humoral immune responses to mycobacterial heat shock protein-65 and its human homologue in Takayasu's arteritis. AB - Expression of heat shock protein (HSP)-65 as well as infiltration of T-cells in arterial lesions and raised levels of circulating antibodies against mycobacterial HSP65 (mHSP65) led us to the concept that mHSP65 or its human homologue (hHSP60) might be involved in the etiopathogenesis of Takayasu's arteritis (TA). Therefore, we investigated mHSP65 and hHSP60 reactive peripheral blood T-cell subsets by BrdU incorporation assay and flow cytometry as well as investigating the different isotypes of anti-mHSP65 and hHSP60 antibodies by ELISA. Eighty-four percent (22/26) of the TA patients were observed to show T cell proliferation to mHSP65 and hHSP60 whereas only 16% (3/18) healthy controls showed such proliferation (P <0.001). Both HSPs induced proliferation of exclusively CD4+ T-cells and not CD8+ T-cells. We also observed a significantly higher prevalence of only the IgG isotype reactive to mHSP65 and hHSP60 in TA as compared to HC (mHSP65: 92% TA versus 11% HC, P <0.0001 and hHSP60: 84% versus 22%, P <0.001). Our data show a significant correlation between mHSP65 and hHSP60 reactive T-cells (CD3+: r=0.901; CD4+: r=0.968) as well as anti-mHSP65 and anti hHSP60 IgG antibodies (r=0.814) suggesting an infection induced autoimmunity in TA, possibly induced by molecular mimicry between mHSP65 and hHSP60 or other tissue specific antigens. PMID- 15544634 TI - Cytokine and chemokine levels in systemic sclerosis: relationship with cutaneous and internal organ involvement. AB - Systemic sclerosis (SSc) is a connective tissue disorder characterized by excessive collagen deposition in the skin and internal organs. Several cytokines and chemokines have been implicated in the induction of fibrosis, but a definitive relationship between specific cytokines and organ involvement has not been established yet. Serum samples, PBMC and T cell lines (TCL) obtained from 54 patients affected by SSc and 20 healthy donors (HD) were examined by ELISA for Interferon-gamma (IFN-gamma ), interleukin (IL)-4, IL-6, IL-10, IL-18, Transforming growth factor (TGF)-beta1, Tumour necrosis factor (TNF)-alpha, sCD30, Macrophage derived chemokine (MDC), Monocyte chemoattractant protein (MCP) 1, Macrophage inflammatory protein (MIP)-1alpha and Regulated on activation normal T-cell expressed and secreted (RANTES). In all the SSc serum samples, we found significantly increased levels of IL6, TNFalpha and MCP-1 but reduced amounts of gamma-IFN and MDC. IL6, IL10, IL18, MIP-1alpha and TNFalpha measured in supernatants from PHA-stimulated PBMC and IL6, MCP-1 and RANTES in supernatants from stimulated TCL were also increased in patients. MDC was decreased in all the biological SSc sources studied. TGF-beta1, IL10, and sCD30 were produced at a significantly lower level by SSc TCL. Serum IL6 and sCD30 levels were significantly increased in dc-SSc patients compared to lc-SSc as were levels of MCP-1 produced by PBMC and IL10 from TCL. We observed a strict relationship between pulmonary fibrosis and IL10, MCP-1 (both from TCL) and serum IL6. Kidney involvement was related to serum MCP-1 levels and IL18 production from PBMC. Oesophageal involvement correlated with MDC production from PBMC and IL10 synthesis by TCL. We showed that IL-6, IL-10, MDC and MCP-1 are variably associated with internal organ involvement and allow the discrimination between limited and diffuse forms of the disease. PMID- 15544637 TI - Global health services research: challenging the future. PMID- 15544638 TI - Health service utilization in the former soviet union: evidence from eight countries. AB - BACKGROUND: In the past decade, the countries that emerged from the Soviet Union have experienced major changes in the inherited Soviet model of health care, which was centrally planned and provided universal, free access to basic care. The underlying principle of universality remains, but coexists with new funding and delivery systems and growing out-of-pocket payments. OBJECTIVE: To examine patterns and determinants of health care utilization, the extent of payment for health care, and the settings in which care is obtained in Armenia, Belarus, Georgia, Kazakhstan, Kyrgyzstan, Moldova, Russia, and Ukraine. METHODS: Data were derived from cross-sectional surveys, representative of adults aged 18 and over in each country, conducted in 2001. Multistage random sample of 18,428 individuals, stratified by region and area, was obtained. Instrument contained extensive data on demographic, economic, and social characteristics, administered face-to-face. The analysis explored the health seeking behavior of users and nonusers (those reporting an episode of illness but not consulting). RESULTS: In the preceding year, over half of all respondents visited a medical professional, ranging from 65.7 percent in Belarus to 24.4 percent in Georgia, mostly at local primary care facilities. Of those reporting an illness, 20.7 percent of all did not consult although they felt they should have done so, varying from 9.4 percent in Belarus to 42.4 percent in Armenia and 49 percent in Georgia. The main reason for not seeking care was lack of money to pay for treatment (45.2 percent), self treatment with home-produced remedies (32.9 percent), and purchase of nonprescribed medicine (21.8 percent). There are marked differences between countries; unaffordability was a particularly common factor in Armenia, Georgia, and Moldova (78 percent, 70 percent, 54 percent), and much lower in Belarus and Russia. In Georgia and Armenia, 65 percent and 56 percent of those who had consulted paid out-of-pocket, in the form of money, gifts, or both; these figures were 8 percent and 19 percent in Belarus and Russia respectively and 31.2 percent overall. The probability of not consulting a health professional when seriously ill was significantly higher among those over age 65, and with lower education. Use of health care was markedly lower among those with fewer household assets or a shortage of money, and those dissatisfied with their material resources, factors that explained some of the effects of age. A lack of social support (formal and informal) decreases further the probability of not consulting, adding to the consequences of poor financial status. The probability of seeking care for common conditions varies widely among countries (persistent fever: 56 percent in Belarus; 16 percent in Armenia) and home remedies, alcohol, and direct purchase of pharmaceuticals are commonly used. Informal coping strategies, such as use of connections (36.7 percent) or offering money to health professionals (28.5 percent) are seen as acceptable. CONCLUSIONS: This article provides the first comparative assessment of inequalities in access to health care in multiple countries of the former Soviet Union, using rigorous methodology. The emerging model across the region is extremely diverse. Some countries (Belarus, Russia) have managed to maintain access for most people, while in others the situation is near collapse (Armenia, Georgia). Access is most problematic in health systems characterized by high levels of payment for care and a breakdown of gate-keeping, although these are seen in countries facing major problems such as economic collapse and, in some, a legacy of civil war. There are substantial inequalities within each country and even where access remains adequate there are concerns about its sustainability. PMID- 15544639 TI - Using vignettes to compare the quality of clinical care variation in economically divergent countries. AB - OBJECTIVE: To determine whether clinical vignettes can measure variations in the quality of clinical care in two economically divergent countries. DATA SOURCE/STUDY SETTING: Primary data collected between February 1997 and February 1998 at two Veterans Affairs facilities in the United States and four government run outpatient facilities in Macedonia. STUDY DESIGN: Randomly selected, eligible Macedonian and U.S. physicians (>97 percent participation rate) completed vignettes for four common outpatient conditions. Responses were judged against a master list of explicit quality criteria and scored as percent correct. DATA COLLECTION/ EXTRACTION: An ANOVA model and two-tailed t-tests were used to compare overall scores by case, study site, and country. Principal Findings. The mean score for U.S. physicians was 67 percent (+/-11 percent) compared to 48 percent (+/-11 percent) for Macedonian physicians. The quality of clinical practice, which emphasizes basic skills, varied greatly in both sites, but more so in Macedonia. However, the top Macedonian physicians in all sites approached or-in one case-exceeded the median score in the U.S. sites. CONCLUSIONS: Vignettes are a useful method for making cross-national comparisons of the quality of care provided in very different settings. The vignette measurements revealed that some physicians in Macedonia performed at a standard comparable to that of their counterparts in the United States, despite the disparity of the two health systems. We infer that in poorer countries, policy that promotes improvements in the quality of clinical practice-not just structural inputs-could lead to rapid improvements in health. PMID- 15544640 TI - Impacts of informal caregiver availability on long-term care expenditures in OECD countries. AB - OBJECTIVE: To quantify the effects of informal caregiver availability and public funding on formal long-term care (LTC) expenditures in developed countries. DATA SOURCE/STUDY SETTING: Secondary data were acquired for 15 Organization for Economic Cooperation and Development (OECD) countries from 1970 to 2000. STUDY DESIGN: Secondary data analysis, applying fixed- and random-effects models to time-series cross-sectional data. Outcome variables are inpatient or home heath LTC expenditures. Key explanatory variables are measures of the availability of informal caregivers, generosity in public funding for formal LTC, and the proportion of the elderly population in the total population. DATA COLLECTION/EXTRACTION METHOD: Aggregated macro data were obtained from OECD Health Data, United Nations Demographic Yearbooks, and U.S. Census Bureau International Data Base. PRINCIPAL FINDINGS: Most of the 15 OECD countries experienced growth in LTC expenditures over the study period. The availability of a spouse caregiver, measured by male-to-female ratio among the elderly, is associated with a $28,840 (1995 U.S. dollars) annual reduction in formal LTC expenditure per additional elderly male. Availability of an adult child caregiver, measured by female labor force participation and full-time/part-time status shift, is associated with a reduction of $310 to $3,830 in LTC expenditures. These impacts on LTC expenditure vary across countries and across time within a country. CONCLUSIONS: The availability of an informal caregiver, particularly a spouse caregiver, is among the most important factors explaining variation in LTC expenditure growth. Long-term care policies should take into account behavioral responses: decreased public funding in LTC may lead working women to leave the labor force to provide more informal care. PMID- 15544641 TI - Assessing the efficiency of HIV prevention around the world: methods of the PANCEA project. AB - OBJECTIVE: To develop data collection methods suitable to obtain data to assess the costs, cost-efficiency, and cost-effectiveness of eight types of HIV prevention programs in five countries. DATA SOURCES/STUDY SETTING: Primary data collection from prevention programs for 2002-2003 and prior years, in Uganda, South Africa, India, Mexico, and Russia. STUDY DESIGN: This study consisted of a retrospective review of HIV prevention programs covering one to several years of data. Key variables include services delivered (outputs), quality indicators, and costs. DATA COLLECTION/EXTRACTION METHODS: Data were collected by trained in country teams during week-long site visits, by reviewing service and financial records and interviewing program managers and clients. PRINCIPAL FINDINGS: Preliminary data suggest that the unit cost of HIV prevention programs may be both higher and more variable than previous studies suggest. CONCLUSIONS: A mix of standard data collection methods can be successfully implemented across different HIV prevention program types and countries. These methods can provide comprehensive services and cost data, which may carry valuable information for the allocation of HIV prevention resources. PMID- 15544642 TI - Private care and public health: do vaccination and prenatal care rates differ between users of private versus public sector care in India? AB - OBJECTIVE: To determine whether patients who use private sector providers for curative services have lower vaccination rates and are less likely to receive prenatal care. DATA SOURCES/STUDY SETTING: This study uses data from the 52d round of the National Sample Survey, a nationally representative socioeconomic and health survey of 120,942 rural and urban Indian households conducted in 1995 1996. STUDY DESIGN: Using logistic regression, we estimate the relationship between receipt of preventive care at any time (vaccinations for children, prenatal care for pregnant women) and use of public or private care for outpatient curative services, controlling for demographics, household socioeconomic status, and state of residence. DATA COLLECTION/EXTRACTION METHODS: We analyzed samples of children ages 0 to 4 and pregnant women who used medical care within a 15-day window prior to the survey. PRINCIPAL FINDINGS: With the exception of measles vaccination, predicted probabilities of the receipt of vaccinations and prenatal care do not differ based on the type of provider at which children and women sought curative care. Children and pregnant women in households who use private care are almost twice as likely to receive preventive care from private sources, but the majority still obtains preventive care from public providers. CONCLUSIONS: We do not find support for the hypothesis that children and pregnant women who use private care are less likely to receive public health services. Results are consistent with the notion that Indian households are able to successfully navigate the coexisting public and private systems, and obtain services selectively from each. However, because the study employed an observational, cross-sectional study design, findings should be interpreted cautiously. PMID- 15544643 TI - To prevent, react, and rebuild: health research and the prevention of genocide. AB - OBJECTIVE: To develop an approach to the primary prevention of genocide, based on established public health-based violence prevention methods derived from a variety of high-risk settings. DATA SOURCES: (1) Peer-reviewed literature in the fields of public health, violence/injury prevention, medicine, economics, sociology, psychology, history, and genocide studies, (2) demographic and health data bases made available by governments and international organizations, (3) reports on recent episodes of genocide published by international and nongovernmental organizations, (4) newspaper and journalistic accounts of recent and past genocides, (5) archival testimonies of genocide victims and perpetrators, and (6) court transcripts of international genocide prosecutions. STUDY DESIGN: The research was conducted as a medical-historical policy analysis synthesizing data within the following framework: (1) Assessment of current violence and injury prevention models for suitability in the prevention of extreme, population-wide violence, (2) analysis of morbidity and mortality data to quantify the impact of genocide on the health of populations, (3) making an inventory of the known societal risk factors for genocidal violence, (4) identification of the theorized, modifiable attitudinal risk factors for genocidal behavior within a population health model, and (5) assessment of existing projects targeting primary violence and injury prevention in high risk jurisdictions, for future adaptation within a structured, public health approach. PRINCIPAL FINDINGS: Mortality rates due to genocidal violence are far in excess of other public health emergencies including malaria and HIV/AIDS. The immediate and long-range health consequences of genocide include the sequelae of infectious diseases, organ system failure, and psychiatric disorders, conferring an increased burden of disease on affected populations for multiple subsequent generations. The impact of genocide on local health economies is catastrophic, and the opportunity costs of diverting scarce global health dollars toward ameliorating genocide related outcomes are substantial. Structural risk factors for genocide within societies include: totalitarian government, exclusionary ideologies, armed conflict, economic hardship, and inaction of bystander nations. Proposed psychological risk factors for genocidal behavior include: moral exclusion, authority orientation, action in self-interest, desensitization, and compartmentalized thinking. Violence and injury prevention models, incorporating what is currently known about the societal and behavioral risk factors for genocide in high-risk populations, may be modified to address the primary prevention of catastrophic violence on a population-wide scale. A number of existent global peace building initiatives may serve as models for the design of future prevention initiatives in high-risk, pre-genocide jurisdictions. CONCLUSIONS: Our analysis suggests that genocide is one of the most pressing threats to the health of populations in the twenty-first century. Recent advances in the public health discipline of violence prevention provide a blueprint for approaches to primary genocide prevention based on epidemiological methods. PMID- 15544644 TI - Franchising reproductive health services. AB - OBJECTIVES: Networks of franchised health establishments, providing a standardized set of services, are being implemented in developing countries. This article examines associations between franchise membership and family planning and reproductive health outcomes for both the member provider and the client. METHODS: Regression models are fitted examining associations between franchise membership and family planning and reproductive health outcomes at the service provider and client levels in three settings. RESULTS: Franchising has a positive association with both general and family planning client volumes, and the number of family planning brands available. Similar associations with franchise membership are not found for reproductive health service outcomes. In some settings, client satisfaction is higher at franchised than other types of health establishments, although the association between franchise membership and client outcomes varies across the settings. CONCLUSIONS: Franchise membership has apparent benefits for both the provider and the client, providing an opportunity to expand access to reproductive health services, although greater attention is needed to shift the focus from family planning to a broader reproductive health context. PMID- 15544645 TI - The impact of a microfinance program on client perceptions of the quality of care provided by private sector midwives in Uganda. AB - OBJECTIVE: To assess the impact of a microfinance program that provided business skills training and revolving loans to private sector midwives on perceived quality of services and client loyalty. STUDY DESIGN: A quasi-experimental study with a pretest, posttest design was used to evaluate the impact of the intervention. Exit interviews were conducted at 15 clinics that received the intervention and 7 clinics that did not. Baseline exit interviews were conducted between November and December 2000. Five days of business skills training were provided to midwives, and loans (averaging $454) were given during January and February 2001. A follow-up clinic visit was made to assess whether midwives were implementing what was emphasized during the training. The loans were to be repaid with interest within 6 to 12 months, at an interest rate that is standard within the local commercial market. For those who repaid the first set of loans (11 clinics), a second set of loans (averaging $742) was provided after June 2001. Follow-up exit interviews were conducted at the same clinics between February and March 2002. We assessed the effect of the intervention at both clinic and client levels. T-tests, the analysis of variance, and multivariate logistic regression analysis were conducted. PRINCIPAL FINDINGS: These findings should be interpreted cautiously since secular trends were observed during the study period. The intervention was associated with improvement in clients' perceptions of the quality of care received at intervention clinics. The intervention was also associated with a higher level of client loyalty. CONCLUSIONS: The enthusiastic response of midwives and the high loan repayment rate indicate that midwives were very receptive to the microfinance program. Overall, these findings suggest that microfinance may have an important role in strengthening private sector health services by increasing private providers' business skills and clients' satisfaction with services. PMID- 15544646 TI - Cost convergence between public and for-profit hospitals under prospective payment and high competition in Taiwan. AB - OBJECTIVE: To test the hypotheses that: (1) average adjusted costs per discharge are higher in high-competition relative to low-competition markets, and (2) increased competition is associated with cost convergence between public and for profit (FP) hospitals for case payment diagnoses, but not for cost-plus reimbursed diagnoses. DATA SOURCES: Taiwan's National Health Insurance database; 325,851 inpatient claims for cesarean section, vaginal delivery, prostatectomy, and thyroidectomy (all case payment), and bronchial asthma and cholelithiasis (both cost-based payment). STUDY DESIGN: Retrospective population-based, cross sectional study. DATA ANALYSIS: Diagnosis-wise regression analyses were done to explore associations between cost per discharge and hospital ownership under high and low competition, adjusted for clinical severity and institutional characteristics. PRINCIPAL FINDINGS: Adjusted costs per discharge are higher for all diagnoses in high-competition markets. For case payment diagnoses, the magnitudes of adjusted cost differences between public and FP hospitals are lower under high competition relative to low competition. This is not so for the cost based diagnoses. CONCLUSIONS: We find that the empirical evidence supports both our hypotheses. PMID- 15544647 TI - Capacity in Thai public hospitals and the production of care for poor and nonpoor patients. AB - OBJECTIVE: To assess the capacity of Thai public hospitals to proportionately expand services to both the poor and the nonpoor. This is accomplished by measuring the production of services provided to poor, relative to nonpoor, patients and the plant capacity of individual public hospitals to care for the patient load. STUDY SETTING: Thai public hospitals operating in 1999, following the economic crisis when public hospitals were required to treat all patients irrespective of ability to pay. STUDY DESIGN AND DATA COLLECTION: Input and output data for 68 hospitals were collected using databases and questionnaire surveys. A distinction was made between inpatient and outpatient services to both poor and nonpoor patients and the data were assessed statistically. DATA ANALYSIS: Congestion and capacity indices to measure poor/nonpoor service trade offs and capacity utilization were estimated. The analysis was undertaken by data envelopment analysis (DEA), a nonparametric linear programming approach used to derive efficiency and productivity estimates. Principal Findings. Increases in the amount of services provided to poor patients did not reduce the amount of services to nonpoor patients. Overall, hospitals are producing services relatively close to their capacity given fixed inputs. Possible increases in capacity utilization amounted to 5 percent of capacity. CONCLUSIONS: Results suggest that some increased public hospital care can be accomplished by reallocation of resources to less highly utilized hospitals, given the budgetary constraints. However, further expansion and increase in access to health services will require plant investments. The study illustrates how DEA methodologies can be used in planning health services in data constrained settings. PMID- 15544648 TI - Effects of global budgeting on the distribution of dentists and use of dental care in Taiwan. AB - OBJECTIVE: To examine the effects of global budgeting on the distribution of dentists and the use and cost of dental care in Taiwan. DATA SOURCES: (1) Monthly dental claim data from January 1996 to December 2001 for the entire insured population in Taiwan. (2) The 1996-2001 population information for the cities, counties and townships in Taiwan, abstracted from the Taiwan-Fukien Demographic Fact Book. STUDY DESIGN: Longitudinal, using the autocorrelation model. PRINCIPAL FINDINGS: Results indicated decline in dental care utilization, particularly after the implementation of dental global budgeting. With few exceptions, dental global budgeting did not improve the distribution of dental care and dentist supply. CONCLUSIONS: The experience of the dental global budget program in Taiwan suggested that dental global budgeting might contain dental care utilization and that several conditions might have to be met in order for the reimbursement system to have effective redistributive impact on dental care and dentist supply. PMID- 15544649 TI - The effect of artificial saliva on the rheological properties of tooth whitening systems. AB - This work was undertaken to explore the effect of saliva addition on the rheological properties of two contrasting tooth bleaching systems, one of which was a paste (Colgate Platinum) and the other a gel (Zaris, 3M ESPE). Using a dynamic stress rheometer with cone and plate geometry, it was shown that addition of artificial saliva reduced the apparent viscosity of each material. However, in some cases this was accompanied by an increase in elasticity. It is suggested that saliva may not have a deleterious effect on the ability of the materials to remain in the bleaching tray. PMID- 15544650 TI - Comparative radical production and cytotoxicity induced by camphorquinone and 9 fluorenone against human pulp fibroblasts. AB - Camphorquinone (CQ) is widely used as a photo-initiator in dental materials; however, its cytotoxicity against human pulp fibroblasts (HPF) and particularly the effects of 2-dimethylaminoethyl methacrylate (DMA), a reducing agent and visible light (VL) irradiation on it remain unknown. So we investigated the cytotoxic and reactive oxygen species (ROS)-producing effects of CQ with or without DMA, in the presence or absence of VL on HPF cells. The free-radical production activity of CQ was measured by two different methods [using diphenylpicryl hydrazyl and galvinoxyl]. The phase-transition properties of dipalmitoylphosphatidyl choline (DPPC) liposomes, as a model for biomembranes, induced by CQ were investigated by differential scanning calorimetry. These findings were compared with those of 9-fluorenone (9F), an aromatic photo initiator with long conjugated groups. Camphorquinone with VL irradiation increased the radical production, whereas 9F with VL irradiation increased ROS production, as well as effecting changes in the DPPC phase-transition properties. The cytotoxicity of CQ towards HPF cells was smaller than that of 9F despite greater radical production. The addition of DMA to the photosensitizer enhanced the free-radical production without increasing the ROS level or the cytotoxicity. Camphorquinone/DMA is a valuable combination for the polymerization of dental resins. PMID- 15544651 TI - Cytotoxic effects of dental resin liquids on primary gingival fibroblasts and periodontal ligament cells in vitro. AB - Cytotoxic effects of resin liquids of three in situ relining dental polymers, Alike, Kooliner, and Tokuso Rebase, and their major components, methyl methacrylate (MMA), isobutyl methacrylate (IBMA), and 1,6-hexanediol dimethacrylate (1,6-HDMA) were investigated. The concentrations of major monomers in these resin liquids were determined by high-performance liquid chromatography. Cellular viability of human gingival fibroblasts (GF) and periodontal ligament (PDL) cells were evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide assay. Moreover, patterns of cell death were analysed using annexin V/propidium iodide staining with flow cytometry. The results indicated that Alike liquid contained 91.3% MMA, Kooliner liquid contained 94.5% IBMA, and Tokuso Rebase liquid contained 65.8% 1,6-HDMA. All materials examined had cytotoxic effects on GF and PDL cells in dose-dependent manners. Tokuso Rebase liquid appeared to be the most cytotoxic among the various resin liquids examined. The effects of Kooliner and Tokuso Rebase liquids may have resulted from IBMA and 1,6-HDMA, respectively. Furthermore, the majority of treated cells died from necrosis; whereas a small portion of cells died from apoptosis. In conclusion, the results demonstrated that these liquid forms of dental polymers and their major monomers cause cytotoxic reactions. The direct relining procedure that cures these materials in situ should be used cautiously. PMID- 15544652 TI - Bonding efficacy and side effects of the high purity glyceryl mono-methacrylate. AB - The purpose of this study was to examine the efficacy of experimentally developed high purity glyceryl mono-methacrylate (GM) as a dentine primer and to evaluate the possibility of allergic reaction. The efficacy of experimental dentine primers was evaluated by measuring the polymerization contraction gap width and the tensile bond strength. Allergic reaction was evaluated by the guinea-pig maximization test. The skin reaction was evaluated according to the criteria of the International Contact Dermatitis Research Group. Contraction gap formation was completely prevented and the tensile bond strength was not significantly affected by priming with GM on high-purity GM. The allergic response decreased when high-purity GM was employed as the challenger. It was concluded that the clinical use of the high-purity GM is beneficial as the delayed allergic reaction could be avoided with no detrimental effect on the dentine bonding. PMID- 15544653 TI - Bond strength of Gradia veneering composite to fibre-reinforced composite. AB - This study investigated the shear bond strength of light-curing veneering composite resin to glass fibre-reinforced composite (FRC). Polymer pre impregnated FRC reinforcement was further impregnated with dimethacrylate monomer resin. The light polymerized FRC substrate was ground and dimethacrylate intermediate resin was applied on the surface before the light-curing veneering composite. Adhesional behaviour of veneering composite to the initially light polymerized FRC substrate was compared with well-polymerized FRC substrate. The treatment time of FRC substrate by the intermediate resin for 5 s and 5 min were also compared. Shear bond strength of veneering composite to FRC was determined for dry and thermocycled specimens (n = 6). The analysis of variance (anova) revealed significant differences (P = 0.042) between the shear bond strengths when 5 s and 5 min intermediate resin treatment times were compared. The highest shear bond strength (21.0 MPa) for FRC substrates was achieved when the well polymerized FRC substrate was treated for 5 min with the intermediate resin and stored dry before tests. Thermocycling reduced the shear bond strengths. The results of this study suggest that applying the intermediate resin increased the shear bond strength values of veneering composite to FRC with multiphase polymer matrix. It was also concluded, that the use of multiphase polymer matrix FRC can be polymerized to high degree of conversion without deferiorating the shear bond strength of veneering composite to the FRC. PMID- 15544654 TI - Effect of 10% sodium ascorbate on the shear bond strength of composite resin to bleached bovine enamel. AB - The purpose of this study was to comparatively investigate the effect of antioxidant treatment and delayed bonding after bleaching with three different concentrations of carbamide peroxide (CP) on the shear bond strength of composite resin to enamel. One hundred flat buccal enamel surfaces obtained from bovine incisors were divided into three bleaching groups of 10, 16 and 22% CP (n = 30) and a control group. Each bleaching group was then divided into three subgroups (n = 10). Group 1 consisted of specimens bonded immediately after bleaching. Group 2 specimens were treated with antioxidant agent, 10% sodium ascorbate, while Group 3 specimens were immersed in artificial saliva for 1 week after bleaching. Specimens in the control group were not bleached. After the specimens were bonded with Clearfil SE Bond and Clearfil AP-X, they were thermocycled and tested in shear until failure. Fracture analysis of the bonded enamel surface was performed using scanning electron microscope. The shear bond strength data was subjected to one-way analysis of variance followed by Duncan's multiple range test at a significance level of P < 0.05. Shear bond strength of composite resin to enamel that was bonded immediately after bleaching with 10, 16 and 22% CP was significantly lower than that of unbleached enamel (P < 0.05). For all three bleaching groups, when the antioxidant-treated and delayed bonding (1 week) subgroups were compared with the control group, no statistically significant differences in shear bond strength were noted (P < 0.05). PMID- 15544655 TI - Microleakage of different cementation techniques in Class V ceramic inlays. AB - Microleakage between the restorative materials and the cavity walls of teeth remains a problem. The purpose of this study was to evaluate the microleakage around Class V ceramic inlay restoration using three different cementation techniques. Class V preparations were made on the buccal surfaces of 45 freshly extracted premolar teeth. The teeth were randomly divided into three groups of 15. In the first group, dentine bonding agent (DBA) (Clearfil Liner Bond 2V) was applied immediately after the tooth preparation (D-DBA). In the second group, DBA was applied to the prepared cavities while luting ceramic inlays (I-DBA). In the third group, DBA was not applied at either stage (No-DBA). Impressions were made and ceramic inlays (Ceramco II) fabricated and cemented in the Class V cavities. The restorations were stored in water at room temperature for 24 h. All restorations were thermocycled and then subjected to a dye penetration test. After sectioning, leakage at cavity/restoration interface was scored. Statistical analysis was performed using Kruskal-Wallis, Mann-Whitney U and Wilcoxon's signed tests. Results showed no significant difference among three different cementation techniques (P > 0.05). Microleakage at the dentine margins was greater than that at the enamel margins (P < 0.05). PMID- 15544656 TI - Surface roughness of flowable and packable composite resin materials after finishing with abrasive discs. AB - The aim of this study was to compare surface roughness of flowable (Admira Flow, Filtek Flow, Tetric Flow) and packable (Admira, Filtek P60, Tetric HB) composite resin restorative materials finishing with Sof-Lex discs by means of average surface roughness (Ra) measurement using a surface profilometer and scanning electron microscopy (SEM). For each test group five specimens were prepared and roughness was measured in five different positions using a profilometer with a traversing distance of 4 mm and a cut-off value of 0.8 mm. The radius of the tracing diamond tip was 5 microm and measuring force and speed was 4 mN and 0.5 mm/s, respectively. The surface roughness of each individual disk was taken as the arithmetic mean of the Ra values measured in five different positions. Additionally, one specimen of each test group after finishing was observed under SEM with the magnification of x800 and x2500. Before finishing with Sof-Lex discs, flowable composite materials showed a smoother surface than packable composites restoratives (P < 0.05). However, after the finishing procedures similar surface textures were observed both from packable and flowable composites (P > 0.05). PMID- 15544657 TI - Application of Heatless Glass as a coating material for denture base-evaluation of denture retentive force. AB - Heatless Glass (HG) was applied to the basal surface of dentures as a coating material. To evaluate the influence of this material on the retentive force of the dentures, the adhesive force of pairs of resin specimens (heat curing resin), metal specimens (Co-Cr alloy), and HG metal specimens (Co-Cr alloy coated with HG), using tap water or resting saliva as a medium was measured, and the following results were obtained. (i) The adhesive force of metal specimens coated with HG material was significantly larger (by 2.3-2.6 times) than that of resin materials (P < 0.001), and it was slightly lower (0.8-0.9 times) than that of metal specimens, showing no significant difference (P > 0.10). (ii) The adhesive force of specimens using tap water as a medium was approximately one-half to two third of that of specimens using resting saliva as a medium. (iii) When resting saliva was used as the medium, the maximum difference in the adhesive force of the specimens among subjects was 1.7 times. (iv) These findings suggested that, from the perspective of adhesive force, HG coating material is suitable for application to the basal surface of dentures. PMID- 15544658 TI - Finite element analysis of fixed partial denture replacement. AB - The purpose of this study was to investigate, by means of the finite element method the mechanical behaviour of three designs of fixed partial denture (FPD) for the replacement of the maxillary first premolar in shortened dental arch therapy. Two-dimensional, linear, static finite element analyses were carried out to investigate the biomechanics of the FPDs and their supporting structures under different scenarios of occlusal loading. Displacement and stress distribution for each design of FPD were examined, with particular attention being paid to the stress variations along the retainer-abutment--and the periodontal ligament-bone interfaces. The results indicated that displacement and maximum principal stresses in the fixed-fixed, three-unit FPD were substantially less than those in the two-unit cantilever FPDs. Of the two cantilever FPDs investigated, the distal cantilever design was found to suffer less displacement and stresses than the mesial cantilever design under similar conditions of loading. The highest values for maximum principal stress in the cantilever FPDs were found within the connector between the pontic and the retainer, and within the periodontal ligament and adjacent bone on the aspect of the retainer away from the pontic. PMID- 15544659 TI - Cover version or re-mix: something old, something new. PMID- 15544660 TI - Newspaper coverage of a violent assault by a mentally ill person. AB - The media is an important source of public information on mental ill-health. A man with a serious psychiatric illness attacked a minister with a knife at a Remembrance Sunday service in a remote, rural part of the Highlands, inflicting a severe facial wound. We aimed to identify lessons for the National Health Service (NHS) from the media coverage of the incident and of a subsequent court case and NHS Highland inquiry and in addition to explore how newspaper reporters approached reporting such incidents. We searched local and regional, national Scottish, and the Scottish editions of three UK newspapers for relevant coverage. We also conducted structured telephone interviews with eight reporters who had attended the inquiry press conference. Most of the media coverage was associated with the assault and the court case, rather than the inquiry results. Only three of 10 inquiry recommendations were mentioned in any reports. Coverage largely dealt with identified shortfalls, rather than proposed solutions. The NHS had made little comment in advance of the announcement of the inquiry results. Most of the newspaper coverage had already occurred. The NHS therefore limited its opportunity to influence newspaper coverage. The interpretation of the results is limited by the size of the study, but the coverage of such events forms part of the discourse on mental health in the media episodes and may have some affect on public perception of mental health issues. We conclude that, without providing confidential information, the NHS should take a more active stance in providing information on the nature and treatment of mental illness in such instances, even in advance of court cases. PMID- 15544661 TI - Impact of health reform on registered psychiatric nursing practice. AB - This paper addresses the impact of health reform on registered psychiatric nursing practice. Over a nine-month period, seven focus groups were conducted with registered psychiatric nurses (RPNs; n = 33) from a variety of practice settings in south central regions of the province of Manitoba located in western Canada. Analysis of data collected from the focus groups are summarized according to the four probing questions utilized in this study, and are discussed in relation to: past registered psychiatric nursing, influential factors, future for registered psychiatric nursing, and proactive strategies. It is acknowledged that while reform has created an increase in the degree and level of independence for RPNs, findings suggest that some RPNs are concerned about insufficient preparation for these new and expanded roles, and all study participants are concerned about the current and future shortage of RPNs. Recommendations include commitment of sufficient funding and employer support for advanced education of RPNs as well as provision of additional funding for increases in the number of available seats for psychiatric nursing education, additional RPN faculty resources and a graduate education programme specifically designed for RPNs. PMID- 15544662 TI - Service users, metaphors and teamworking in mental health. AB - Despite the United Kingdom's recent governmental mental health policy directives aimed at strengthening professional collaboration and increasing service user involvement, the prevailing mental health care culture remains steeped in a discourse of treatment and care, control and compliance and professional expertise. Drawing upon the data collected during the two phases of a 2-year national evaluation undertaken for the English National Board for Nursing, Midwifery and Health Visiting, the perceptions of a group of mental health service users in relation to their experiences and contact with the multi professional team are explored. A series of metaphorical descriptions were developed with these service users drawn from their experience. These begin to illuminate a realistic way of thinking about how teams are set up, how and why they carry out their various roles, and the need to think in non-professional terms about the relationships that are developed with service users. A hierarchy of power was noted that was congruent with the outcomes of other studies. However, there was also a concurrent acknowledgement of the 'usefulness'(to the individual service user) of each of the professional group members. This appeared to have been constructed alongside the power hierarchy and serves to illustrate how individual service users sought to find an accommodation within the social system they were placed in. This paper argues however, that the use of metaphors, as a form of shared communication, can be an effective first step in working towards this objective. Working in the way described here can allow for a greater shared understanding of what each group is experiencing and help ensure that future service development reflects a broader view of the mental health care world. PMID- 15544663 TI - The use and nursing management of benzodiazepines in acute, mental health inpatient care: a discussion. AB - The use of medication in acute mental health is common practice however, there is a lack of research into all aspects of pharmacology within inpatient settings. This paper explores the specific use of benzodiazepines and areas of potential mismanagement. A number of priorities for investigation are raised and implications for nursing roles and responsibilities discussed. PMID- 15544664 TI - Understanding treatment adherence in affective disorders: a qualitative study. AB - The objective of this study was to explore the perspective of depressed patients, their family and mental health professionals from their community mental health centre (CMHC) on factors related to treatment adherence. We conducted eight separate focus groups involving patients, their families and their therapists from three CMHCs. A total of 52 persons were involved. The groups explored patients' and family's explanatory model of depression, perceptions about the course of the disease, the role of medication and other treatments, the main causes of non-adherence, and interventions which would help increase it. Patients and families had a complex cognitive model of depression, which combined intrinsic vulnerability, psychological suffering during childhood and adolescence, and adverse life events. Drugs as well as other treatments were considered helpful, more so by patients than by family members. Denial of the disease and need to test its continuing presence were the main causes of non adherence for patients, while adverse reactions did not play a relevant role. Mental health professionals tended to underestimate non-adherence in depressed patients, and did not question their patients about medication adherence. Family members needed more information on depression, on how to manage their relatives, as well as psychological and social support for themselves. The study allowed for the identification of a number of specific interventions aimed at facilitating treatment adherence, such as providing more information to patients and families, and training doctors and nurses in effective prevention and management of non adherence. PMID- 15544665 TI - The coping of an adolescent who has been in psychiatric inpatient care and her mother in everyday life. AB - The purpose of this study was to describe the coping of an adolescent girl who had been psychiatric inpatient care and her mother in everyday life. The case study design was used to gain intimate knowledge of the persons' condition, thoughts, feelings, actions, intentions and environment. The data were collected by interviewing the adolescent and her mother during 1999-2001. The informants were interviewed three times. The data were analysed by inductive content analysis. Six categories were formulated to describe the coping demands faced by the adolescent who had been in psychiatric inpatient care. They were: suicidal behaviour, failing mental health, narrow social network, weakening sense of reality, low self-esteem and negative feedback from others. Her coping strategies were hobbies, praying and occasionally withdrawal into a fantasy world. Emotional support was her most important coping resource. The coping demands faced by the mother concerned her daughter's aggressive behaviour, withdrawal from personal relationships and inability to concentrate. The mother worried about her daughter's unstable mental health and future. She was also extremely concerned about the relationships between the family members. On top of everything else, the mother developed new somatic diseases. The mother took care of herself by studying and doing handiwork. Social life was very important to her. Her main coping resource was emotional support. She got emotional support from friends and members of the family. The family had previously kept in touch with friends, but the husband had broken off all contacts. PMID- 15544666 TI - Perspectives of a nurse, a social worker and a psychiatrist regarding patient assessment in acute inpatient psychiatry settings: a case study approach. AB - This case study explores what informs and organizes the assessment of patients, as undertaken by a nurse, a social worker and a psychiatrist in public, metropolitan, acute mental health service settings. The research data are the transcripts of in-depth interviews with three experienced practitioners, one from each of the three disciplines. The analysis draws on Foucauldian concepts: discourse as constructed through practices of discipline and the gaze. We explored examples of taken-for-granted assessment practices and their interplay with discourse. The findings suggest that participating practitioners use language in assessment in ways that support the powerful discourses of the professional disciplines. The competing discourse of management, associated with industry and economics, is evident in hospital admission processes, dictating the times and places of assessment. Professional and management discourses both effectively marginalize the perspective of another player in assessment, the patient. PMID- 15544667 TI - Specialist drug misuse nurse's motivation, clinical decision-making and professional communication: an exploratory study. AB - UK guidelines advocate a multidisciplinary shared care approach to manage drug related problems. While general practitioners (GPs), consultants and pharmacists have been researched, there has been little research into the role of nurses in this field. This study examined nurses specialized in the field of substance misuse. It specifically explored their motivation for entering this area, interaction with other health professionals and their role in clinical decision making. Seventeen face-to-face interviews were undertaken with nurses from the study population of the 21 nursing staff currently employed in one service. Responses were tape-recorded, transcribed and analysed using qualitative methods. The majority of nurses entered this field because of a specific interest in its patients and their problems. Various professionals interacted with nurses: specialist medical staff, who served as an expert resource and pharmacists through the dispensing of substitute drugs. GP participation in services was inconsistent forcing some nurses to take undue responsibility for clinical decision-making. It is essential that nurses entering this field have an interest in this field and positive attitudes towards drug users, and greater emphasis should be placed on this attribute when selecting new employees. If a shared care model is to be successful, it is important that all GPs accept their responsibilities and relieve pressure currently placed on specialist services. Alternatively, it may be necessary to adopt a new model of care. PMID- 15544668 TI - Link Family Support - an evaluation of an in-home support service. AB - A significant body of research has shown that parents who have a child or a dependent adult with a disability experience significant and persistent levels of stress. One of the recognized strategies for coping includes the provision of in home practical support. Enable Ireland provides a range of services for children and adults with a physical disability. The present study explored home support services with a sample of 16 families of service users of Enable Ireland Cork. Practical support was deemed to be whatever support or intervention requested by the parent of the child/dependent adult which afforded the service user the opportunity to engage in social/recreational opportunities and that gave the parent free time. Sixteen members of the chosen families were interviewed (15 mothers and one father) using a semi-structured interview schedule and a standardized stress measure before and after the introduction of Link Family Support (LFS). LFS was put in place for a period of 12 months, tailored to the families and service users' individual needs. Although levels of stress continued to be high and scores on the Parenting Stress Index (PSI) did not show a statistically significant reduction after the programme, reported stress levels had improved. Parents reported LFS to be very helpful in reducing perceived stress and improving family's quality of life through providing free time and access to leisure and recreation facilities. This study provided limited but clear evidence of the need for regular, flexible, in-home support for families with children and dependent adults with a disability. LFS provided a personal, regular and effective means of meeting this need as the findings of this study demonstrated. PMID- 15544669 TI - Perceptions on the standardization of psychiatric work: development of a care pathway. AB - Policy development and practice for hospital care has shifted towards a more deterministic approach. Crucial within this development is the assumption that interventions can be standardized to fit within the confines of a practice framework. A system to deliver standardized approaches to care is a care pathway. A research study was carried out to determine how care could be standardized to fit within a care pathway for people diagnosed with schizophrenia. A range of interviews with the multidisciplinary team and observations of the working group process was the data collection technique. Analysis was driven by emergent themes across the data set. Clinicians expressed divergent views on the nature of standardized care for people with schizophrenia. Respondents also offered a deeper understanding of standardized care by arguing for a sense of flexibility to be built into care systems. Findings from this study have implications for the introduction of NICE core standards for people with schizophrenia. For example, clinicians may be so opposed to such structures as to work against them. By understanding different views on standardized care, it may ultimately support their introduction into practice. PMID- 15544670 TI - Mental health student nurses' perception of the role of the mental health nurse. AB - Clear role definition is essential for directing the focus of nurse education and several studies have attempted to define the role of the mental health nurse (MHN). These, however, came to the conclusion that mental health nursing was difficult to articulate. The aim of this study was to understand how, during their transition to first level registration, mental health student nurses (MHSNs) perceived the role of the MHN. Semi-structured interviews were conducted with 14 MHSNs during the last 6 months of their transition to MHN. Transcripts were analysed using a qualitative approach based on grounded theory. Six key themes were identified, five of which were defined mental health nursing roles. The sixth theme related to non-therapeutic intervention on the part of some MHNs. Several areas of concern were identified. First, MHSNs expected to conduct more psychologically based interventions than were achievable in practice. Second, emphasis on drug administration can lead to a conflict of interest in the nurses' advocacy role with patients. Third, MHSNs sometimes observed poor role models in their placements, which could have a negative impact on the way in which future MHNs view the role of the MHN. PMID- 15544671 TI - Quality of life: a basis for clinical decision-making in community psychiatric care. AB - This descriptive study used grounded theory methodology to illuminate how the concept of quality of life influences the practice of community psychiatric nurses caring for individuals with serious mental illnesses. In-depth interviews captured the practice wisdom of expert nurses. Data were verified through focus groups, member checks and peer debriefing. The constant comparative method was used to categorize data and identify key themes and concepts. Informants regarded the concept of quality of life as central to nursing practice. They cited examples of how it serves as a goal and philosophy of care, a basis for forming relationships, and a lens through which the nursing role becomes clearer. Quality of life is acknowledged to be a subjective concept. It is assessed in light of the individual's hopes, dreams and values, and within the context of his or her whole life. A quality of life focus is supported by the philosophies of holism and empowerment. Nurses promote quality of life through symptom management, educating and advocating for clients. Minimizing barriers to quality of life and helping the individual to have a broader vision of what his or her life could be, are also central to the psychiatric nursing role. PMID- 15544672 TI - The reality of practice development for nurses working in an inpatient service for people with severe and enduring mental health problems. AB - Nursing practice development is a growing priority for the British National Health Service. However, the nature of practice development remains poorly articulated. Despite the growing number of papers on practice development in nursing, there is insufficient guidance in the literature for the practical day to-day management of the role, particularly in the inpatient environment. This paper explores the tensions experienced by practitioners engaged in practice development within a service for people mainly with a diagnosis of psychosis. The entrenched culture of the environment is described, which was resistant to change therefore did not easily embrace practice development. Within such a culture it is important to balance effective management with team development and support. Although this balance is essential for positive change, it is the maintenance of the balance that often leads to the tensions experienced by practice development practitioners. The paper describes numerous tensions including the competing agendas, influencing 'hearts and minds' rather than forcing change and working where people are at. Each tension is explored alongside possible survival strategies. The material presented aims to be realistic and although it may not be generalizable to other settings and staff groups, the authors hope to stimulate debate about similar or contrasting experiences of practice development. PMID- 15544673 TI - Identity and the experience of postnatal depression: the use of Video Interaction Guidance. PMID- 15544677 TI - Ignoring the evidence dictating the practice: sexual orientation, suicidality and the dichotomy of the mental health nurse. PMID- 15544680 TI - Trust and shared decision-making: an emerging research agenda. PMID- 15544681 TI - A community study using specified and unspecified scenarios to investigate men's views about PSA screening. AB - OBJECTIVE: We explored the influence of different but factual scenarios about prostate-specific antigen (PSA) screening on men's interest in having PSA screening to detect early prostate cancer. DESIGN: Cross-sectional, representative community survey. SETTING AND PARTICIPANTS: A total of 514 men (89% response fraction) aged 50-70 years randomly selected from a telephone directory database in Sydney, Australia. MAIN VARIABLES STUDIED: Demographic, health and psychological variables. MAIN OUTCOME VARIABLES: Interest in undergoing screening in response to five unspecified scenarios and, elsewhere in our interview, a specified scenario in which PSA screening was mentioned explicitly. RESULTS: When presented with a scenario describing a lack of evidence underpinning the efficacy of screening for an unspecified cancer, 61.2% of men reported that they 'probably' or 'definitely' wanted to undergo screening for an unspecified cancer. Similar proportions reported that they 'probably' or 'definitely' wanted to undergo screening even at the risk of unmasking indolent cancer (60.9%) or without expert consensus about the value of screening (62.8%). Greatest interest in screening was elicited in that scenario describing life-time risk of dying from prostate cancer (72.6%) (P < 0.001). Significantly fewer indicated they would 'probably' or 'definitely' want to undergo screening for a cancer for which there was uncertainty about treatment efficacy and known side effects (46.1%) (P < 0.001). Increasing age was a consistent predictor of positive interest in screening. When asked later in our survey specifically about PSA screening, 68.1%'probably' or definitely' wanted PSA screening. CONCLUSION: Public health policy makers need to ensure that men are provided with the scope of medical evidence germane to prostate cancer screening and treatment, thereby potentially improving prostate cancer screening decisions. PMID- 15544682 TI - Grounded citizens' juries: a tool for health activism? AB - Involving the public in decision-making has become a bureaucratic pre-occupation for every health agency in the UK. In this paper we offer an innovative approach for local participation in health decision-making through the development of a 'grounded' citizens' jury. We describe the process of one such jury commissioned by a Primary Care Group in the north-west of England, which was located in an area suffering intractable health inequalities. Twelve local people aged between 17 and 70 were recruited to come together for a week to hear evidence, ask questions and debate what they felt would improve the health and well-being of people living in the area. The jury process acted effectively as a grass-roots health needs assessment and amongst other outcomes, resulted in the setting up of a community health centre run by a board consisting of members of the community (including two jurors) together with local agencies. The methodology described here contrasts with that practiced by what we term 'the consultation industry', which is primarily interested in the use of fixed models to generate the public view as a standardized output, a product, developed to serve the needs of an established policy process, with little interest in effecting change. We outline four principles underpinning our approach: deliberation, integration, sustainability and accountability. We argue that citizens' juries and other consultation initiatives need to be reclaimed from that which merely serves the policy process and become 'grounded', a tool for activism, in which local people are agents in the development of policies affecting their lives. PMID- 15544683 TI - Education and role modelling for clinical decisions with female cancer patients. AB - BACKGROUND: Patients vary widely in their preferences and capacity for participating in treatment decision-making. There are few interventions targeting patient understanding of how doctors make decisions and shared decision-making. This randomized trial investigates the effects of providing cancer patients with a package designed to facilitate shared decision-making prior to seeing their oncologist. PATIENTS AND METHODS: Sixty-five female cancer patients were randomized to receive either the package (booklet and 15-min video) or a booklet on living with cancer, before their initial consultation. Participants completed questionnaires prior to the intervention, immediately after the oncology consultation, and 2 weeks and 6 months later. The first consultation with the oncologist was audio-taped and transcribed. RESULTS: Patients receiving the package were more likely than controls to declare their information and treatment preferences in the consultation, and their perspectives on the costs, side effects and benefits of treatment. Doctors introduced considerably more new themes in the consultations with intervention subjects than they did with controls; no other differences in doctor behaviour were noted. CONCLUSIONS: This short intervention successfully shifted patient and doctor behaviour closer to the shared decision-making model, although it did not alter patients' preferences for information or involvement. PMID- 15544684 TI - How does trust affect patient preferences for participation in decision-making? AB - OBJECTIVE: Does trust in physicians aid or hinder patient autonomy? We examine the relationship between trust in the recipient's doctor, and desire for a participative role in decisions about medical treatment. DESIGN: We conducted a cross-sectional survey in an urban Canadian teaching hospital. SETTING AND PARTICIPANTS: A total of 606 respondents in three clinics (breast cancer, prostate cancer, fracture) completed questionnaires. VARIABLES STUDIED: The instrument included the Problem Solving Decision Making (PSDM) Scale, which used two vignettes (current health condition, chest pain) to categorize respondents by preferred role, and the Trust-in-Physician Scale. RESULTS: Few respondents preferred an autonomous role (2.9% for the current health condition vignette and 1.2% for the chest pain vignette); most preferred shared decision-making (DM) (67.3% current health condition; 48.7% chest pain) or a passive role (29.6% current health condition; 50.1% chest pain). Trust-in-physician yielded 6.3% with blind trust, 36.1% with high trust, 48.6% moderate trust and 9.0% low trust. As hypothesized, autonomous patients had relatively low levels of trust, passive respondents were more likely to have blind trust, while shared respondents had high but not excessive trust. Trust had a significant influence on preferred role even after controlling for the demographic factors such as sex, age and education. CONCLUSIONS: Very few respondents wish an autonomous role; those who do tend to have lower trust in their providers. Familiarity with a clinical condition increases desire for a shared (as opposed to passive) role. Shared DM often accompanies, and may require, a trusting patient-physician relationship. PMID- 15544685 TI - A randomized trial among women with heavy menstruation -- impact of a decision aid on treatment outcomes and costs. AB - OBJECTIVE: To evaluate the effects of a decision aid for menorrhagia on treatment outcomes and costs over a 12-month follow-up. DESIGN: Randomized trial and pre trial prospective cohort study. SETTING AND PARTICIPANTS: Gynaecology outpatient clinics in 14 Finnish hospitals, 363 (randomized trial) plus 206 (cohort study) patients with menorrhagia. INTERVENTION: A decision aid booklet explaining menorrhagia and treatment options, mailed to patients before their first clinic appointment. MAIN OUTCOME MEASURES: Health related quality of life, psychological well-being, menstrual symptoms, satisfaction with treatment outcome, use and cost of health care services. RESULTS: All study groups experienced overall improvement in health-related quality of life, anxiety, and psychosomatic and menstrual symptoms, but not in sexual life. Treatment in the intervention group was more active than in the control group, with more frequent course of medication and less undecided treatments. However, there were no marked disparities in health outcomes, satisfaction with treatment outcome and costs. Total costs (including productivity loss) per woman because of menorrhagia over the 12-month follow-up were 2760 and 3094 in the intervention and control group, respectively (P = 0.1). The pre-trial group also had a significantly lower rate of uterus saving surgery compared with the control group, but no difference in costs because of menorrhagia treatment. CONCLUSION: Despite some differences in treatment courses, a decision aid for menorrhagia in booklet form did not increase the use of health services or treatment costs, nor had it impact on health outcomes or satisfaction with outcome of treatment. PMID- 15544686 TI - Self-reported use of shared decision-making among breast cancer specialists and perceived barriers and facilitators to implementing this approach. AB - BACKGROUND: Physicians are increasingly urged to practice shared decision-making with their patients. Using a cross-sectional survey, we explored the extent to which Ontario breast cancer specialists report practising shared decision-making with their patients, their comfort level with this approach, and perceived barriers and facilitators to implementation. PARTICIPANTS AND METHODS: All Ontario surgeons and oncologists (radiation and medical) treating women with early-stage breast cancer were eligible for this study. Likert scales were used to measure physicians' comfort level with and self-reported use of different treatment decision-making approaches as well as perceived barriers and facilitators to treatment decision-making with patients. RESULTS: The response rate was 79% for oncologists and 72% for surgeons. More physicians from each specialty (87% of oncologists and 89% of surgeons) expressed high levels of comfort with clinical example 4 (designed to illustrate a shared approach) than with any of the other examples presented (e.g. the informed and paternalistic approach). Similarly, more oncologists and surgeons reported that their usual approach to treatment decision-making was like example 4 than like any other approach presented (56% of oncologists and 69% of surgeons, respectively). Comfort levels with example 4 for oncologists and surgeons were 31% and 20% higher, respectively, than the reported use of this approach. Lack of time and patient anxiety, patient lack of information and/or misinformation, and patient unwillingness or inability to participate were perceived by a substantial minority of both oncologists and surgeons as barriers to patient involvement in treatment decision-making. Key facilitators identified included patients' emotional readiness, support, information and trust in the physician. More research is needed to identify contextual, physician, patient, and interaction factors that will facilitate shared decision-making in the medical encounter and help both parties create an environment conducive to implementing this approach to the extent desired. PMID- 15544687 TI - Developing a learning network to support engagement with health communities. PMID- 15544689 TI - HIV and nanobacteria. PMID- 15544690 TI - Effect of concurrent zidovudine use on the resistance pathway selected by abacavir-containing regimens. AB - OBJECTIVES: Abacavir (ABC) selects for four mutations (K65R, L74V, Y115F and M184V) in HIV-1 reverse transcriptase (RT), both in vitro and during monotherapy in vivo. The aim of this analysis was to compare the selection of these and other nucleoside reverse transcriptase inhibitor (NRTI)-associated mutations by ABC containing therapies in the presence and absence of concurrent lamivudine (3TC) and/or zidovudine (ZDV) and to assess the effect of these mutations on phenotypic susceptibility to the NRTIs. DESIGN: This study was a retrospective analysis of the patterns of NRTI-associated mutations selected following virological failure in six multicentre trials conducted during the development of ABC. METHODS: Virological failure was defined as confirmed vRNA above 400 HIV-1 RNA copies/mL. RT genotype and phenotype were determined using standard methods. RESULTS: K65R was selected infrequently by ABC-containing regimens in the absence of ZDV (13 of 127 patients), while L74V/I was selected more frequently (51 of 127 patients). Selection of both K65R and L74V/I was significantly reduced by co-administration of ZDV with ABC (one of 86 and two of 86 patients, respectively). Y115F was uncommon in the absence (seven of 127 patients) or presence (four of 86 patients) of ZDV. M184V was the most frequently selected mutation by ABC alone (24 of 70 patients) and by ABC plus 3TC (48 of 70 patients). Thymidine analogue mutations were associated with ZDV use. The K65R mutation conferred the broadest phenotypic cross-resistance of the mutations studied. CONCLUSIONS: The resistance pathway selected upon virological failure of ABC-containing regimens is significantly altered by concurrent ZDV use, but not by concurrent 3TC use. These data may have important implications for the efficacy of subsequent lines of NRTI therapies. PMID- 15544691 TI - An algorithm-based genotypic resistance score is associated with clinical outcome in HIV-1-infected adults on antiretroviral therapy. AB - OBJECTIVES: The aim of this study was to evaluate the association between genotypic drug resistance and the occurrence of HIV-related diseases and death in HIV-1-infected adults on antiretroviral therapy. METHODS: We performed an observational study on patients from an out-patient clinic in a university hospital. Genotypic drug resistance analysis after virological treatment failure was performed in 141 patients receiving two or more antiretroviral drugs. All patients had follow up of at least 6 months after the resistance test. An algorithm was developed to estimate the level of genotypic drug resistance and to assign an actual resistance score (ARS) for the drugs prescribed to each patient. The patient population was divided into quartiles according to patients' ARS values. Our endpoint was the risk of developing an HIV-related disease [Centers for Disease Control and Prevention (CDC) category B or C] during the period starting 6 months prior to and ending 6 months after the genotypic resistance test, or death during the 6 months following the resistance test. RESULTS: There was a significant association between the level of resistance to the drugs prescribed (ARS) and our clinical endpoint: the odds ratio for an endpoint (with 95% confidence interval) was 3.20 (1.28-7.99), adjusted for CD4 cell count and HIV RNA, in patients in the highest ARS quartile compared with patients in the other three quartiles. CONCLUSIONS: Our study indicates that patients with high level genotypic drug resistance are at increased risk of developing an HIV related disease. This association could not be explained by differences in CD4 cell count or HIV RNA levels. PMID- 15544692 TI - Restoration of CD4 T-cell responses to cytomegalovirus is short-lived in severely immunodeficient HIV-infected patients responding to highly active antiretroviral therapy. AB - OBJECTIVES: To define the level of pathogen-specific immune reconstitution persisting over 3 to 5 years of highly active antiretroviral therapy (HAART) in HIV-infected patients who began therapy with CD4 T-cell counts below 50 cells/microL. METHODS: Cytomegalovirus (CMV)-specific T-cell responses were analysed in adult HIV-1-infected patients with nadir CD4 T-cell counts below 50 cells/microL before HAART. CMV-specific CD4 T-cell responses were measured by interferon-gamma enzyme-linked immunospot assay (ELISpot assay), lymphoproliferation and interferon-gamma levels in cell culture supernatants. RESULTS: CD4 T-cell responses to CMV were low in untreated patients and remained low during the first year on HAART, but increased progressively to levels similar to those found in HIV-seronegative CMV-seropositive controls at 3 years. Responses then declined markedly and at 5 years were lower than controls. This could not be explained by changes in CD4 or CD8 T-cell counts or plasma HIV RNA levels. Interferon-gamma and interleukin-5 responses to a mitogen were maintained or elevated. CONCLUSIONS: CMV-specific CD4 T-cell responses were found to decline after 3-5 years on HAART and may provide inadequate long-term protection against CMV disease in patients who are severely immunodeficient prior to treatment. PMID- 15544693 TI - Findings from the British HIV Association's national clinical audit of first-line antiretroviral therapy and survey of treatment practice and maternity care, 2002. AB - OBJECTIVES: To assess the impact of and adherence to the British HIV Association (BHIVA) guidelines in routine clinical practice in the UK and Ireland. METHODS: A self-completed postal survey of practice with regard to HIV management and maternity care of HIV-positive women was carried out. A case-note review of all patients starting their first antiretroviral (ART) therapy (up to a maximum of 25 patients) between April and September 2002 was also performed. RESULTS: A total of 113 treatment centres participated. Ninety-nine units (88%) use the BHIVA ART guidelines as part of their treatment policy. Problems revealed by the survey include limited access to some ARTs in a small number of units and a wait longer than 4 weeks before the first assessment of patients started on ART in some units. Thirty-one centres (33%) felt that their maternity services had reached the government target of >90% antenatal HIV testing by October 2002. A total of 942 case-notes audited showed shortfalls in reported pretreatment testing of blood glucose, lipids and hepatitis C virus. Three patients had not strictly met the BHIVA guidelines for starting ART, but all had a high viral load (>100 000 HIV-1 RNA copies/mL). The majority (844 patients; 90%) had started on recommended therapies, most commonly two nucleoside reverse transcriptase inhibitors and a nonnucleoside reverse transcriptase inhibitor. The other 98 patients (10%) were given a variety of nonstandard triple or quadruple ART. CONCLUSIONS: This audit again shows a high rate of consistency and quality of care of patients with HIV across the UK and Ireland. Problems with pretreatment assessment and early follow up of patients starting ART have been found. PMID- 15544694 TI - Symptomatic bone disorders in HIV-infected patients: incidence in the Aquitaine cohort (1999-2002). AB - BACKGROUND: Since the inception of highly active antiretroviral therapy (HAART), mortality among HIV-infected patients has decreased, but this has been accompanied by the appearance of several complications. OBJECTIVES: To estimate the incidence of symptomatic bone disorders in HIV-infected patients of the Aquitaine cohort (from south-west France) for the period 1999-2002, and to describe cases. METHODS: We retrospectively studied the records of 2700 patients of the Aquitaine cohort, which was derived from a hospital-based surveillance system of HIV infection in France. All cases of symptomatic bone disorders diagnosed from 1 January 1999 to 30 June 2002 were reviewed. RESULTS: Fourteen cases of bone disorders were diagnosed, eight cases of aseptic osteonecrosis and six cases of severe osteoporosis, representing incidences of 0.3/1000 patient years [95% confidence interval (CI): 0.14-0.62] and 0.22/1000 patient-years (95% CI: 0.09-0.52), respectively. All patients with aseptic osteonecrosis were male, while all but one with osteoporosis were female. The ages of patients ranged from 36 to 54 years for osteonecrosis and from 39 to 50 for severe osteoporosis. At the time of clinical diagnosis, all patients were treated with nucleoside reverse transcriptase inhibitors (duration of treatment ranging from 19 to 123 months for osteonecrosis and from 46 to 132 months for severe osteoporosis). Ten patients were treated with nonnucleoside reverse transcriptase inhibitors [duration of treatment ranging from 6 to 31 months for osteonecrosis (n=6) and from 4 to 29 months for severe osteoporosis (n=4)]. Thirteen patients were treated with protease inhibitors [duration of treatment ranging from 12 to 62 months for osteonecrosis (n=8) and from 3 to 44 months for severe osteoporosis (n=5)]. All osteonecrosis and five osteoporosis patients had at least one known risk factor or comorbidity associated with the bone disorder occurrence. CONCLUSIONS: In our study, the aetiology of clinical bone disorders seemed to be multifactorial, as almost all the patients had at least one possible risk factor in addition to HAART exposure. PMID- 15544695 TI - GB virus C coinfection and vertical transmission in HIV-infected mothers before the introduction of antiretroviral prophylaxis. AB - OBJECTIVES: To investigate the prevalence of GB virus C (GBV-C) viraemia and GBV C antibodies in a cohort of HIV-infected mothers and their infants between 1987 and 1994. METHODS: GBV-C viraemia and antibodies were determined by polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) in 52 HIV infected mothers and their 53 infants, who were born before antiretroviral prophylaxis for reduction of HIV transmission was introduced at the end of 1994. Ten of these children acquired HIV. RESULTS: Mothers of three children had GBV-C viraemia and mothers of another 14 children carried antibodies against GBV-C. No mother had GBV-C antibodies and GBV-C viraemia simultaneously. GBV-C viraemia was detected in only one infant. This child was delivered by the vaginal route to a mother with GBV-C viraemia, and was not HIV-infected. No vertical transmission of GBV-C occurred from mothers with GBV-C antibodies. However, four of 10 children who were infected with HIV had a mother with past or ongoing GBV-C infection. CONCLUSION: Our findings suggest that the risk of vertical transmission of GBV-C is not elevated in HIV-infected mothers. Furthermore, although the number of HIV 1-infected children was low, we saw no evidence that the presence of ongoing or past GBV-C infection influenced the probability of vertical HIV transmission. PMID- 15544696 TI - Epidemiological, clinical and therapeutic features of AIDS-related Mycobacterium kansasii infection during the HIV pandemic: an 11-year follow-up study. AB - OBJECTIVES: Optimal diagnosis and timely treatment of atypical mycobacteriosis, and especially Mycobacterium kansasii disease, remain a serious challenge for clinicians engaged in the management of the immunocompromised host. METHODS AND RESULTS: From more than 2700 hospitalizations (over 1800 patients) attributable to HIV-associated disorders over an 11-year period, 12 patients were found to have a confirmed M. kansasii infection. This reflects the recent reduction in the frequency of this HIV-related complication, which virtually disappeared after the introduction of potent antiretroviral combinations in 1996. In the early 1990s, the lack of effective antiretroviral regimens made frequent the association with AIDS, a mean CD4 lymphocyte count of nearly 20 cells/microL, and an extremely variable chest X-ray features. The recent detection of a further case was attributable to late recognition of very advanced HIV disease, complicated by multiple opportunistic disorders. CONCLUSIONS: Mycobacterium kansasii respiratory or disseminated infection continues to occur, and poses diagnostic problems in terms of late or missed identification as a result of slow culture and frequently concurrent opportunistic disease. Serious therapeutic difficulties also arise from the unpredictable in vitro antimicrobial susceptibility profile of these organisms, and from the need to start an effective combination therapy that does not interfere with other medications as soon as possible. PMID- 15544697 TI - Changes in hospital admissions across Europe: 1995-2003. Results from the EuroSIDA study. AB - OBJECTIVES: To describe changes in the proportions of patients admitted to hospital and the duration of admission during the month of March between 1995 and 2003 and to describe the factors related to admission for 9802 patients from EuroSIDA, a pan-European, observational cohort study. METHODS: Generalized estimating equations were used to determine changes over time in the proportion of patients admitted and the median duration of admission. Logistic regression was used to determine factors related to admission in March 1995, March 1998 and March 2001. RESULTS: The proportion of patients admitted during March declined from 7.4% in 1995 to 2.6% in 2003. After adjustment, the estimated reduction in the proportion of patients admitted was 5.5% per year [95% confidence interval (CI) 2.5-8.5%; P=0.0004], a 26% reduction. The median duration of hospital admission declined by 58% from 12 days in 1995 [interquartile range (IQR) 5-19 days] to 5 days in 2003 (IQR 3-12 days), a significant decline of 0.7 days per year after adjustment (95% CI 0.5-0.9 days; P=0.031). Patients with a lower CD4 lymphocyte count, and with an AIDS diagnosis made within the 3 months prior to March, all had increased odds of admission during March 1995, 1998 or 2001. In March 2001, patients whose treatment regimen was changed as a consequence of toxicities had increased odds of admission [odds ratio (OR) 2.34; 95% CI 1.26 4.37; P=0.0074]. In addition, patients who were hepatitis C virus-positive during March 2001 (OR 1.66; 95% CI 1.02-2.68; P=0.041) had increased odds of admission. CONCLUSIONS: There has been a considerable decline in both the proportion of patients admitted to hospital and the median duration of the stay. Patients with hepatitis C had increased odds of admission, but there was little evidence of an increase in admissions among patients taking highly active antiretroviral therapy (HAART) associated with serious adverse events, although longer follow up is required. PMID- 15544698 TI - Bilateral enophthalmos as a manifestation of HIV infection-related lipoatrophy. PMID- 15544699 TI - Tenofovir use in a patient with a severe renal impairment. PMID- 15544700 TI - Merkel cell carcinoma and multiple basal cell carcinoma in an African albino woman with HIV infection. AB - A 25-year-old HIV-infected African albino woman developed an aggressive Merkel cell carcinoma on her face and at least 10 basal cell carcinomas, mainly on sun exposed parts of her body. HIV infection, immune deficiency and sun exposure are known risk factors for the development of Merkel cell carcinoma. Chemotherapy and radiotherapy were only temporarily successful. She died shortly after surgery was performed to remove the tumour. PMID- 15544701 TI - Family physicians' perspectives on practice guidelines related to cancer control. AB - BACKGROUND: Family physicians (FPs) play an important role in cancer control. While FPs' attitudes towards, and use of guidelines in general have been explored, no study has looked at the needs of FPs with respect to guidelines for the continuum of cancer control. The objective of this study was to understand which guideline topics FPs consider important. METHODS: Five group interviews were conducted by telephone with FPs from across Ontario, Canada. Transcripts were analyzed inductively. Content analysis identified emergent themes. Themes are illustrated by representative quotes taken from the transcripts. RESULTS: The main areas where FPs felt guidelines were needed most included screening - a traditional area of responsibility for FPs - and treatment and follow-up - areas where they felt they lacked the knowledge to best support patients. Confusion over best practice when faced with conflicting guidelines varied according to disease site. FPs defined good guideline formats; the most often cited forms of presentation were tear-off sheets to use interactively with patients, or a binder. Computer-based dissemination was acknowledged as the best way of widely distributing material that needs frequent updates. However, until computer use is a common aspect of practice, mail was considered the most viable method of dissemination. Guidelines designed for use by patients were supported by FPs. CONCLUSIONS: Preferred guideline topics, format, dissemination methods and role of patient guidelines identified by FPs in this study reflect the nature of their practice situations. Guideline developers and those supporting use of evidence based guidelines (e.g., Canadian Strategy for Cancer Control) have a responsibility to ensure that FPs are provided with the resources they identify as important, and to provide them in a format that will best support their use. PMID- 15544703 TI - Increase of malaria attacks among children presenting concomitant infection by Schistosoma mansoni in Senegal. AB - Helminthic infections concomitant with malaria are common in inter-tropical areas. A recent study showed that mice co-infected with Schistosoma mansoni and Plasmodium chabaudi develop higher P. chabaudi parasitaemia and had a higher mortality rate. This important observation deserved to be further investigated among human populations.Malaria attacks were recorded in 512 children aged 6-15 years living in Richard Toll (Northern Senegal) among whom 336 were infected by S. mansoni, and 175 were not. The incidence rate of malaria attacks was significantly higher among S. mansoni-infected individuals, particularly those carrying the highest worm loads, as compared to uninfected subjects (26.6% versus 16,4 %). In contrast, the rate of malaria attacks was lower, without reaching significance, in medium grade S. mansoni infections. Thus, infection by S. mansoni affects susceptibility to malaria, but this can vary according to the intensity of parasite load. The immunological mechanisms underlying this dual effect need to be further explored. PMID- 15544702 TI - Neuroblastoma and pre-B lymphoma cells share expression of key transcription factors but display tissue restricted target gene expression. AB - BACKGROUND: Transcription factors are frequently involved in the process of cellular transformation, and many malignancies are characterized by a distinct genetic event affecting a specific transcription factor. This probably reflects a tissue specific ability of transcription factors to contribute to the generation of cancer but very little is known about the precise mechanisms that governs these restricted effects. METHODS: To investigate this selectivity in target gene activation we compared the overall gene expression patterns by micro-array analysis and expression of target genes for the transcription factor EBF in lymphoma and neuroblastoma cells by RT-PCR. The presence of transcription factors in the different model cell lines was further investigated by EMSA analysis. RESULTS: In pre-B cells mb-1 and CD19 are regulate by EBF-1 in collaboration with Pax-5 and E-proteins. We here show that neuroblastoma cells express these three, for B cell development crucial transcription factors, but nevertheless fail to express detectable levels of their known target genes. Expression of mb-1 could, however, be induced in neuroblastoma cells after disruption of the chromatin structure by treatment with 5-azacytidine and Trichostatin A. CONCLUSION: These data suggest that transcription factors are able to selectively activate target genes in different tissues and that chromatin structure plays a key role in the regulation of this activity. PMID- 15544704 TI - Primary lymphocutaneous nocardiosis in an immunocompetent patient. AB - BACKGROUND: Nocardia brasiliensis is a rare human pathogen usually associated with localized cutaneous infections. CASE PRESENTATION: We report a case of primary lymphocutaneous Nocardia brasiliensis infection developed after a bone fracture of the left hand of an otherwise healthy 32-year-old man. Treatment with trimethoprim-sulfamethoxazole given for a total of three months combined with surgical debridement resulted in complete resolution of the infection. CONCLUSION: Nocardiosis should be part of the differential diagnosis in patients with sporotrichoid infection, particularly those with a history of outdoor injury. Culture of the affected tissue and antimicrobial susceptibility testing of the isolate should be performed for diagnosis and treatment. PMID- 15544706 TI - Platelet-derived NO slows thrombus growth on a collagen type III surface. AB - Nitric oxide (NO) is a free radical that plays an important role in modulating platelet adhesion and aggregation. Platelets are a source of vascular NO, but since erythrocytes avidly scavenge NO, the functional significance of platelet derived NO is not clear. Our purpose was to determine if NO from platelets affects platelet thrombus formation in the presence of anticoagulated whole blood in an in vitro parallel plate flow system. We studied platelet adhesion and aggregation on a collagen type III surface in the presence of physiologically relevant fluid mechanical shear stress. We found that certain receptor mediated agonists (insulin and isoproterenol) caused a concentration dependent reduction in thrombus formation at a shear rate of 1000 s-1. This effect was mediated by NO since it was abolished in the presence of the NO inhibitor L-nitro-arginine methyl-ester (L-NAME). As expected, at venous levels of shear rate (100 s-1) neither of the agonists had any effect on thrombus formation since platelet adhesion does not depend on activation at these low levels of shear. Interestingly, at a shear rate of 2000 s-1 the addition of L-NAME caused an increase in platelet coverage suggesting that shear, by itself, induces NO production by platelets. This is the first demonstration of shear stress causing platelets to produce an inhibitor of platelet activation. These results demonstrate that the development of a platelet thrombus is regulated in a complex way and that platelets produce functionally significant amounts of NO even in the presence of whole blood. PMID- 15544705 TI - Effects of heparin on the uptake of lipoprotein lipase in rat liver. AB - BACKGROUND: Lipoprotein lipase (LPL) is anchored at the vascular endothelium through interaction with heparan sulfate. It is not known how this enzyme is turned over but it has been suggested that it is slowly released into blood and then taken up and degraded in the liver. Heparin releases the enzyme into the circulating blood. Several lines of evidence indicate that this leads to accelerated flux of LPL to the liver and a temporary depletion of the enzyme in peripheral tissues. RESULTS: Rat livers were found to contain substantial amounts of LPL, most of which was catalytically inactive. After injection of heparin, LPL mass in liver increased for at least an hour. LPL activity also increased, but not in proportion to mass, indicating that the lipase soon lost its activity after being bound/taken up in the liver. To further study the uptake, bovine LPL was labeled with 125I and injected. Already two min after injection about 33 % of the injected lipase was in the liver where it initially located along sinusoids. With time the immunostaining shifted to the hepatocytes, became granular and then faded, indicating internalization and degradation. When heparin was injected before the lipase, the initial immunostaining along sinusoids was weaker, whereas staining over Kupffer cells was enhanced. When the lipase was converted to inactive before injection, the fraction taken up in the liver increased and the lipase located mainly to the Kupffer cells. CONCLUSIONS: This study shows that there are heparin-insensitive binding sites for LPL on both hepatocytes and Kupffer cells. The latter may be the same sites as those that mediate uptake of inactive LPL. The results support the hypothesis that turnover of endothelial LPL occurs in part by transport to and degradation in the liver, and that this transport is accelerated after injection of heparin. PMID- 15544707 TI - Proteomics computational analyses suggest that the carboxyl terminal glycoproteins of Bunyaviruses are class II viral fusion protein (beta penetrenes). AB - The Bunyaviridae family of enveloped RNA viruses includes five genuses, orthobunyaviruses, hantaviruses, phleboviruses, nairoviruses and tospoviruses. It has not been determined which Bunyavirus protein mediates virion:cell membrane fusion. Class II viral fusion proteins (beta-penetrenes), encoded by members of the Alphaviridae and Flaviviridae, are comprised of three antiparallel beta sheet domains with an internal fusion peptide located at the end of domain II. Proteomics computational analyses indicate that the carboxyl terminal glycoprotein (Gc) encoded by Sandfly fever virus (SAN), a phlebovirus, has a significant amino acid sequence similarity with envelope protein 1 (E1), the class II fusion protein of Sindbis virus (SIN), an Alphavirus. Similar sequences and common structural/functional motifs, including domains with a high propensity to interface with bilayer membranes, are located collinearly in SAN Gc and SIN E1. Gc encoded by members of each Bunyavirus genus share several sequence and structural motifs. These results suggest that Gc of Bunyaviridae, and similar proteins of Tenuiviruses and a group of Caenorhabditis elegans retroviruses, are class II viral fusion proteins. Comparisons of divergent viral fusion proteins can reveal features essential for virion:cell fusion, and suggest drug and vaccine strategies. PMID- 15544708 TI - Generalisability in economic evaluation studies in healthcare: a review and case studies. AB - OBJECTIVES: To review, and to develop further, the methods used to assess and to increase the generalisability of economic evaluation studies. DATA SOURCES: Electronic databases. REVIEW METHODS: Methodological studies relating to economic evaluation in healthcare were searched. This included electronic searches of a range of databases, including PREMEDLINE, MEDLINE, EMBASE and EconLit, and manual searches of key journals. The case studies of a decision analytic model involved highlighting specific features of previously published economic studies related to generalisability and location-related variability. The case-study involving the secondary analysis of cost-effectiveness analyses was based on the secondary analysis of three economic studies using data from randomised trials. RESULTS: The factor most frequently cited as generating variability in economic results between locations was the unit costs associated with particular resources. In the context of studies based on the analysis of patient-level data, regression analysis has been advocated as a means of looking at variability in economic results across locations. These methods have generally accepted that some components of resource use and outcomes are exchangeable across locations. Recent studies have also explored, in cost-effectiveness analysis, the use of tests of heterogeneity similar to those used in clinical evaluation in trials. The decision analytic model has been the main means by which cost-effectiveness has been adapted from trial to non-trial locations. Most models have focused on changes to the cost side of the analysis, but it is clear that the effectiveness side may also need to be adapted between locations. There have been weaknesses in some aspects of the reporting in applied cost-effectiveness studies. These may limit decision-makers' ability to judge the relevance of a study to their specific situations. The case study demonstrated the potential value of multilevel modelling (MLM). Where clustering exists by location (e.g. centre or country), MLM can facilitate correct estimates of the uncertainty in cost effectiveness results, and also a means of estimating location-specific cost effectiveness. The review of applied economic studies based on decision analytic models showed that few studies were explicit about their target decision maker(s)/jurisdictions. The studies in the review generally made more effort to ensure that their cost inputs were specific to their target jurisdiction than their effectiveness parameters. Standard sensitivity analysis was the main way of dealing with uncertainty in the models, although few studies looked explicitly at variability between locations. The modelling case study illustrated how effectiveness and cost data can be made location-specific. In particular, on the effectiveness side, the example showed the separation of location-specific baseline events and pooled estimates of relative treatment effect, where the latter are assumed exchangeable across locations. CONCLUSIONS: A large number of factors are mentioned in the literature that might be expected to generate variation in the cost-effectiveness of healthcare interventions across locations. Several papers have demonstrated differences in the volume and cost of resource use between locations, but few studies have looked at variability in outcomes. In applied trial-based cost-effectiveness studies, few studies provide sufficient evidence for decision-makers to establish the relevance or to adjust the results of the study to their location of interest. Very few studies utilised statistical methods formally to assess the variability in results between locations. In applied economic studies based on decision models, most studies either stated their target decision-maker/jurisdiction or provided sufficient information from which this could be inferred. There was a greater tendency to ensure that cost inputs were specific to the target jurisdiction than clinical parameters. Methods to assess generalisability and variability in economic evaluation studies have been discussed extensively in the literature relating to both trial-based and modelling studies. Regression-based methods are likely to offer a systematic approach to quantifying variability in patient-level data. In particular, MLM has the potential to facilitate estimates of cost-effectiveness, which both reflect the variation in costs and outcomes between locations and also enable the consistency of cost-effectiveness estimates between locations to be assessed directly. Decision analytic models will retain an important role in adapting the results of cost-effectiveness studies between locations. Recommendations for further research include: the development of methods of evidence synthesis which model the exchangeability of data across locations and allow for the additional uncertainty in this process; assessment of alternative approaches to specifying multilevel models to the analysis of cost-effectiveness data alongside multilocation randomised trials; identification of a range of appropriate covariates relating to locations (e.g. hospitals) in multilevel models; and further assessment of the role of econometric methods (e.g. selection models) for cost-effectiveness analysis alongside observational datasets, and to increase the generalisability of randomised trials. PMID- 15544709 TI - Topical vapocoolant quickly and effectively reduces vaccine-associated pain: results of a randomized, single-blinded, placebo-controlled study. AB - BACKGROUND: Comprehensive international travel preparation often requires several vaccines. Up to 90% of adults have some fear of injections, mostly due to injection-related pain. Pediatric studies with routine vaccines have shown topical anesthetic EMLA cream (lidocaine and prilocaine, Astra Pharmaceuticals, Inc.) and the topical vapocoolant Fluori-Methane (dichlorodifluoromethane and trichlorodifluoromethane, Gebauer Co.) to be equally effective in reducing pain from vaccinations. EMLA cream is expensive and requires a 60-min application, while Fluori-Methane (FM) is immediate in onset of action and inexpensive. Skin anesthesia begins at 10 degrees C. Fluori-Methane can briefly cool the skin to 0 degrees C. METHODS: We studied the effectiveness of topical vapocoolant on adult clients at our international travel clinic in a randomized, controlled trial of topical FM vs. cold (4 degrees C) saline placebo. Using a preset randomization table, participants served as their own controls, receiving placebo/control or active agent (participant blinded) in one arm (left or right), and a similar number of vaccines in the untreated arm. Vaccines were administered according to a set protocol per arm to minimize the risk of bias. Pain was measured using a modified McGill present pain intensity (PPI) pain index. Subjects rated their pain immediately and at 5 min on a six-level scale, noting treated and untreated arms separately. A questionnaire was completed on intervention preferences. Sample size was predetermined to achieve 90% statistical power estimating 25% efficacy (minimum n=172). RESULTS: One hundred and eighty-five participants were enrolled; 93 FM and 92 cold saline placebo. FM-treated arms had a significant reduction in immediate pain compared to untreated arms (pain scale mean 2.2 vs. 3.1; p<.0001), and compared to placebo (mean 2.2 vs. 2.8; p<.01). Delayed pain at 5 min was not affected by FM or control (mean 1.9 vs. 2.0) compared to no intervention (pain scale 1.9). The intervention preference questionnaire indicated that participants did not find FM therapy uncomfortable. They would choose FM therapy in the future, over a cream, especially if a wait was involved. CONCLUSION: The topical vapocoolant Fluori-Methane is an effective, quick, preferred, inexpensive agent for reducing vaccine-associated injection pain for international travel clients. PMID- 15544710 TI - Screening for fever by remote-sensing infrared thermographic camera. AB - BACKGROUND: Following the severe acute respiratory syndrome (SARS) outbreak, remote-sensing infrared thermography (IRT) has been advocated as a possible means of screening for fever in travelers at airports and border crossings, but its applicability has not been established. We therefore set out to evaluate (1) the feasibility of IRT imaging to identify subjects with fever, and (2) the optimal instrumental configuration and validity for such testing. METHODS: Over a 20-day inclusive period, 176 subjects (49 hospital inpatients without SARS or suspected SARS, 99 health clinic attendees and 28 healthy volunteers) were recruited. Remotely sensed IRT readings were obtained from various parts of the front and side of the face (at distances of 1.5 and 0.5 m), and compared to concurrently determined body temperature measurements using conventional means (aural tympanic IRT and oral mercury thermometry). The resulting data were submitted to linear regression/correlation and sensitivity analyses. All recruits gave prior informed consent and our Faculty Institutional Review Board approved the protocol. RESULTS: Optimal correlations were found between conventionally measured body temperatures and IRT readings from (1) the front of the face at 1.5m with the mouth open (r=0.80), (2) the ear at 0.5 m (r=0.79), and (3) the side of the face at 1.5m (r=0.76). Average IRT readings from the forehead and elsewhere were 1 degrees C to 2 degrees C lower and correlated less well. Ear IRT readings at 0.5 m yielded the narrowest confidence intervals and could be used to predict conventional body temperature readings of < or = 38 degrees C with a sensitivity and specificity of 83% and 88% respectively. CONCLUSIONS: IRT readings from the side of the face, especially from the ear at 0.5 m, yielded the most reliable, precise and consistent estimates of conventionally determined body temperatures. Our results have important implications for walk-through IRT scanning/screening systems at airports and border crossings, particularly as the point prevalence of fever in such subjects would be very low. PMID- 15544711 TI - The safety experience of New Zealand adventure tourism operators. AB - BACKGROUND: This survey examined parameters of the New Zealand adventure tourism industry client injury risk. The research also sought to establish priorities for intervention to reduce adventure tourism risk, and identify client injury control measures currently in place (or absent) in the New Zealand adventure tourism industry, with a view to establishing guidelines for the development of effective adventure tourism safety management systems. This 2003 survey builds upon an exploratory study of New Zealand adventure tourism safety conducted by us during 1999. METHOD: A postal questionnaire was used to survey all identifiable New Zealand adventure tourism operators. The questionnaire asked respondents about their recorded client injury experience, perceptions of client injury risk factors, safety management practices, and barriers to safety. RESULTS: Some 27 adventure tourism activities were represented among the responding sample (n=96). The highest client injury risk was reported in the snow sports, bungee jumping and horse riding sectors, although serious underreporting of minor injuries was evident across the industry. Slips, trips and falls (STF) were the major client injury mechanisms, and a range of risk factors for client injuries were identified. Safety management measures were inconsistently applied across the industry. CONCLUSIONS: The industry should consider the implications of poor injury reporting standards and safety management practices generally. Specifically, the industry should consider risk management that focuses on minor (e.g., STF) as well as catastrophic events. PMID- 15544712 TI - Nationwide survey of the role of travel medicine in primary care in Germany. AB - BACKGROUND: With more persons traveling to (sub)tropical destinations, travel medicine is an increasing challenge for the health care system in many countries. This study investigated the role and scope of travel medicine provided by general practitioners (GPs) in primary health care, assessed the determinants of specific prophylactic recommendations, and identified the needs of GPs regarding training and cooperation in travel medicine. METHODS: A national survey was conducted among randomly selected GPs in Germany. Information on the type and extent of activities in travel medicine, the number and characteristics of patients, the use of specific information systems, and training and cooperation, was obtained with a standardized postal questionnaire. Representativeness was assessed by a computer-assisted telephone survey among nonrespondents. Univariate and multivariate statistical analysis was performed. RESULTS: Of all GPs (n=1,320), 95% provided medical pretravel advice or post-travel counseling (13.2 patients/month). The most common topics in pretravel consultation were immunizations (95%), malaria chemoprophylaxis (94%), advice on exposure prophylaxis (41%), and advice on protection from sexually transmitted infections (STIs) (43%). GPs frequently advised certain patient groups (e.g., cardiovascular patients 68%, pregnant women 20%). Travelers' diarrhea was the most common topic in post-travel consultation (89%), followed by fever (38%). Of the GPs, 25% cooperated regularly with institutions with special expertise in travel medicine or tropical diseases. Sixty-four percent had received some training in travel medicine, and 86% stated a need for additional training. In the multivariate analysis, independent determinants for advice on exposure prophylaxis and STI risks included special training in travel medicine, use of country-specific information sources, and location of the practice in the former West Germany and urban areas. CONCLUSIONS: Travel medicine is of importance in primary health care in Germany. There is a need for improved counseling on exposure and STI prophylaxis. GPs' training in travel medicine and cooperation with institutions such as institutes, clinics and laboratories for tropical medicine should be improved. PMID- 15544713 TI - A description of travel medicine in general practice: a postal questionnaire survey. AB - BACKGROUND: Travel-related diseases are important aspects of public health. The number of UK residents traveling abroad is increasing at a rate of 16% a year, thereby increasing exposure to travel-related morbidity. Provision of comprehensive pretravel health advice is essential to reduce this trend. In the UK, pretravel health advice is predominantly provided through general practices. METHODS: A postal questionnaire was sent to all 91 general practices in South Cheshire Health Authority. The questionnaires were to be completed by the lead advisor in travel medicine for each practice. Questions were asked on service provision, training and reference resources used, subjects advised on, and health promotion material used. Nonresponders were contacted and sent a further questionnaire. RESULTS: A response rate of 86% (78/91) was achieved. Of the lead advisors, 97% were nurses and 3% general practitioners. Thirty-eight sources of advice were quoted, the commonest of which comprised wall immunization charts (72%). Duration of consultation ranged from less than 5 min to over 30 min, with a median and mode of 11 to 15 min. Most respondents reported advising on most travel-associated risks, 40% of practices lacked a protocol, and 83% of providers had attended a training course on travel medicine for 2 days or less. CONCLUSIONS: This survey identified inadequacies of training and use of multiple sources of reference which may lead to inconsistencies in advice. Most practitioners could not define their workload in travel medicine. For effective protection of travelers, a careful risk assessment, clear risk communication and health education with detailed health promotion are necessary, but these are not likely to be provided within an average consultation time of 11 to 15 min. There is no evidence of consistent governance, planned training and monitoring of service quality of travel medicine practice. This may be due to lack of a national policy on best practice and guidance in this subject. National protocols with validated information resources, set standards of training, along with adequate consultation time for educating, advising, and prescribing, will lead to improved health of the traveling public. PMID- 15544714 TI - Vacation at moderate and low altitude improves perceived health in individuals with metabolic syndrome. AB - BACKGROUND: Recent data suggest that vacation may improve cardiovascular health, an effect possibly moderated by altitude. The aim of the present study was to study the effect of a 3-week vacation at moderate and low altitude on perceived health in individuals with increased cardiovascular risk. METHODS: Seventy-two overweight males, both occupationally active and retired (mean age=56.6 +/- 7.2 years), with signs of metabolic syndrome were randomly assigned to identical sojourns at either moderate (1,700 m) or low (300 m) altitude and engaged in four 3- to 4-h heart-rate-controlled hiking tours per week. Perceived health was measured 2 weeks before vacation, at the beginning and end of vacation, and 7 weeks after vacation. RESULTS: Fitness, recreational ability, positive and negative mood and social activities improved during vacation, independent of altitude and occupational status, although the day-to-day improvement in quality of sleep was delayed at moderate altitude. During the follow-up examinations, improvements in all reported aspects of health except for social activities were maintained. In comparison to retired individuals, active individuals showed a greater long-term improvement in social activities. CONCLUSION: Vacation positively affects perceived health independent of altitude or occupational status in generally inactive overweight males. PMID- 15544715 TI - Risk and prevention of tick-borne encephalitis in travelers. PMID- 15544716 TI - Urban medicine: threats to health of travelers to developing world cities. PMID- 15544717 TI - Itraconazole can be effective in the treatment of sporotrichoid leishmaniasis. PMID- 15544718 TI - Mycetoma of the foot caused by Cylindrocarpon lichenicola in an immunocompetent traveler. AB - Cylindrocarpon is a cosmopolitan soil fungus, which rarely causes human disease. It has infrequently been reported as causing keratitis, mycetoma, osteomyelitis and peritonitis in chronic peritoneal ambulatory dialysis patients and disseminated infection in leukemic neutropenic hosts. This report describes a case of invasive infection caused by Cylindrocarpon lichenicola, localized in the right foot of an otherwise immunocompetent traveler. PMID- 15544719 TI - Selected bibliography. PMID- 15544722 TI - IMPACT position paper: SNE. PMID- 15544725 TI - Factors of fruit and vegetable intake by race, gender, and age among young adolescents. AB - OBJECTIVE: To explore demographic differences in individual, social, and environmental factors potentially related to fruit and vegetable intake. DESIGN: Self-report questionnaires administered to a convenience sample of middle school students during regular classes. PARTICIPANTS: Black and white adolescents, 11 to 15 years of age (N = 736). VARIABLES MEASURED: Measures included self-efficacy, family dinner frequency, normative beliefs, outcome expectations, modeling, availability, preferences, snack choice, and demographics. ANALYSIS: Chi-square, general linear models, and Poisson and linear regressions as appropriate. RESULTS: Black participants reported greater social influences than did white participants, whereas white adolescents reported greater family environmental influences on fruit and vegetable intake. The oldest adolescents reported lower self-efficacy, peer modeling, family dinner frequency, and fruit and vegetable preferences compared with younger adolescents. White participants and females reported a higher preference for vegetables than did black participants and males. Regression models for self-efficacy and snack choice explained 41% and 34% of the variance, respectively. Preferences for vegetables and parental modeling were the strongest correlates of self-efficacy. Self-efficacy was the strongest correlate of snack choice. CONCLUSIONS AND IMPLICATIONS: Decreases in several factors with age highlight the importance of intervention for this age group. Future research is needed for a better understanding of the formation and modification of self-efficacy and snack choice. PMID- 15544726 TI - Eating after breast cancer: influences on women's actions. AB - OBJECTIVE: To explore women's dietary actions after a breast cancer diagnosis and the factors influencing those actions. DESIGN: Individual interviews exploring women's perceptions. SETTING: Vancouver, British Columbia, Canada. PARTICIPANTS: A culturally diverse convenience sample of women (N = 30) aged 40 to 60 years diagnosed with breast cancer 6 months to 15 years previously. PHENOMENON OF INTEREST: Social and personal factors that influence dietary actions. ANALYSIS: Verbatim interview transcripts were coded and themes developed by sorting and summarizing coded transcript segments. RESULTS: Women held a wide range of beliefs concerning the relationship between diet and breast cancer, particularly their own cancer occurrence. Actions were not always consistent with professed beliefs. Some women believed that diet contributes to breast cancer, even their own cancer, yet made no diet changes; others did not believe in such a relationship but did change their diets postdiagnosis. Family support, employment, financial resources, cultural food patterns, and other health concerns were strong influences. CONCLUSIONS AND IMPLICATIONS: Although beliefs about diet-cancer connections affect actions concerning diet change, the relationship is not consistent. Diet change is also affected by social, cultural, and economic concerns that are at least as important as a woman's beliefs. Nutrient supplements appear to be a common, although perhaps temporary, response to diagnosis. Future research should explore the possibilities for nutrition professionals to help women think critically about the range of influences on their diets. PMID- 15544727 TI - Holocaust survivors report long-term effects on attitudes toward food. AB - OBJECTIVE: To gather information from survivors on the effects that the Holocaust had on their current attitudes toward food. DESIGN: Qualitative study: one-on-one semistructured interviews with a script shaped by a pilot study. SETTING: South Florida homes and community sites, including the Miami Beach Holocaust Memorial. PARTICIPANTS: Convenience sample of 25 Holocaust survivors: 14 men, 11 women; ages 71 to 85 years. PHENOMENON OF INTEREST: Current attitudes toward food in relation to Holocaust experiences. ANALYSIS: Themes and illustrative quotations from transcriptions of audiotaped interviews. RESULTS: Food attitudes were influenced by Holocaust experiences. Five themes emerged: (1) difficulty throwing food away, even when spoiled; (2) storing excess food; (3) craving certain food(s); (4) difficulty standing in line for food; and (5) experiencing anxiety when food is not readily available. Empathy for those currently suffering from hunger was also reported. CONCLUSIONS AND IMPLICATIONS: Food-related issues from the Holocaust remain for survivors. Now in their 70s and 80s, many use health care and related services. Nutritionists, educators, and health professionals should be aware of such issues. Food and nutrition programs should minimize uncomfortable food-related situations for Holocaust survivors and others who experienced food deprivation. PMID- 15544728 TI - Cooking classes outperform cooking demonstrations for college sophomores. AB - OBJECTIVE: To determine if cooking classes improve subjects' knowledge, attitudes, and behaviors toward cooking. DESIGN: Comparison of outcomes of 2 treatment groups (demonstration vs hands-on cooking classes) using pre- and posttests. SETTING: University cooking laboratories. PARTICIPANTS: First-semester sophomores (n = 65) who were 25% male with a mean age of 19.7 years. INTERVENTION: The intervention group (n = 33) attended 4 2-hour cooking classes, based on Social Learning Theory, and a supermarket tour. The demonstration group (n = 32) attended a cooking demonstration. Subjects completed 6 surveys. MAIN OUTCOME MEASURES: Changes in attitudes, knowledge, and behaviors regarding cooking. ANALYSIS: Descriptive statistics were used to compare demographic variables. Analysis of covariance and chi-square analyses were used to compare outcome variables. RESULTS: Analysis revealed no gender differences. Participants displayed positive shifts on attitude scales. The intervention group had a pattern of larger positive shifts (0.4-0.7 vs 0.1-0.5 gain), some of which were statistically significant. Participants displayed positive, but not statistically significant, shifts in knowledge and some behaviors. CONCLUSION AND IMPLICATIONS: The intervention group experienced more statistically significant gains in attitudes and appeared to have a better pattern of gains in cooking-related knowledge and behaviors. Given limited resources, demonstration cooking classes could reach larger audiences in varied settings, but the impact would likely be weaker than that of cooking classes. PMID- 15544729 TI - Soy intervention in adolescent girls: design and implementation. AB - The purpose of this study was to explore the feasibility of implementing a soy intervention in female adolescents. Twenty girls, ages 8 to 14, were recruited to consume 1 daily serving of soymilk or soy nuts. They also provided 9 weekly urine samples over a 2-month period. Information about the study foods and procedures was collected through post-study questionnaires. Adherence to the intervention was successful using strategies that addressed both girls' and mothers' needs. The use of conveniently packaged soy foods, activities to maintain motivation, and frequent contact maintained participation. Future studies should also consider the maturity and sense of responsibility of participants to achieve compliance. PMID- 15544730 TI - Omega-3 fatty acid nutrition education resources. PMID- 15544731 TI - Improving self-awareness of nutrition and lifestyle practices through on-line journaling. PMID- 15544732 TI - Use of the dietary analysis plus program in an introductory college nutrition class. PMID- 15544734 TI - [Sedation in digestive endoscopy. Results of a hospital survey in Catalonia (Spain)]. AB - INTRODUCTION: The need for sedation is increasing in digestive endoscopy units (DEU). There are no data on the use of sedation in DEU in Catalonia (Spain). OBJECTIVE: To evaluate the use of sedation in DEU in Catalonia. MATERIAL AND METHOD: A questionnaire on the practice of sedation was designed and sent to the heads of medical and nursing staff of the DEU of 63 public and private hospitals in Catalonia. Two mailings were sent with an interval of three months between each. The questionnaire included 62 items on the characteristics of the hospital and the DEU, number of explorations, frequency of sedation use, drugs employed, participation of an anesthesiologist, use of monitoring, and complications. RESULTS: Forty-four DEU (70%) corresponding to 31 public hospitals and 13 private hospitals completed the questionnaire. Evaluation of sedation patterns was based on 105,904 explorations performed in the various DEU (56,453 gastroscopies, 47,278 colonoscopies and 2,173 endoscopic retrograde cholangiopancreatographies (ERCP) in 2001. Sedation, sedation-analgesia or anesthesia was used in 17% of gastroscopies, 61% of colonoscopies and 100% of ERCP. Sedation was administered by an anesthesiologist in 7% of gastroscopies, 25% of colonoscopies and 38% of ERCP. Anesthesiologist administration was more frequent in private than in public centers (gastroscopies: 25% vs. 2%; colonoscopies: 57% vs. 9%, p < 0.001). No deaths associated with the use of sedation were reported. Eighty-nine percent of the DEU complied with standard recommendations for the practice of sedation. CONCLUSIONS: In Catalonia, the use of sedation is highly variable, depending on the endoscopic procedure and the DEU. Use of sedation in infrequent in gastroscopy, fairly widespread in colonoscopy and routine in ERCP. Anesthesiologist administration is significantly more frequent in private hospitals. Most DEU follow standard sedation practices. PMID- 15544735 TI - [Study of patients referred for elevated ferritin levels and/or transferrin saturation: significance of non-alcoholic fatty liver disease]. AB - OBJECTIVE: To determine the etiology of increased ferritin concentrations and/or transferrin saturation in patients in whom classical causes were ruled out. PATIENTS AND METHOD: We studied 43 patients (35 males and 8 females) who were referred for ferritinemia greater than 300 ng/ml and or a transferrin saturation index (TSI) greater than 40%. In all patients, glycemia, cholesterol, triglycerides, uric acid, total and fractionated bilirubin, transaminase, gammaglutamyltranspeptidase, sideremia, TSI, ferritin, HFE gene mutations, ceruloplasmin and total 24-hour urine porphyrin were evaluated and abdominal ultrasonography was performed. In 14 patients liver biopsy was performed. RESULTS: Fifty-three percent was overweight and 19% was obese. Alterations in carbohydrate metabolism were detected in 33%, hypercholesterolemia was found in 14%, hypertriglyceridemia in 35%, and hyperlipemia type IIb in 16%. Thirty-two percent showed isolated elevated ferritin, 12% had elevated TSI and 56% showed elevation of both. Transaminase levels were normal in 61%. No mutation in the HFE gene was found in 10 patients, the H63D/wt mutation was found in 18, C262Y/wt in 1, C282Y/H63D in 5, C282Y/C282Y in 4, H63D/H63D in 3 and Ser65cys/wt in 1. Ultrasonography revealed steatosis in 19 patients (44%). Definitive diagnoses were HFE-linked hemochromatosis (4 patients), juvenile hemochromatosis (1 patient), hepaticocutaneous porphyria (1 patient), and non-alcoholic fatty liver disease (22 patients; 51%). Most of the remaining patients could be included under insulin resistance syndrome. Phlebotomy was performed in 25 patients, with improvement in clinical and laboratory parameters. CONCLUSIONS: Non-alcoholic fatty acid disease is frequently detected in patients with iron metabolism disorders. These patients should undergo investigations for metabolic alterations and liver ultrasonography and, if necessary, biopsy. Phlebotomy can be useful in the treatment of these patients. PMID- 15544736 TI - [Determination of hepatitis B virus genotype and detection of lamivudine resistance mutations]. AB - OBJECTIVES: To determine hepatitis B virus (HBV) genotypes in southern Seville (Spain) and investigate the development of lamivudine-resistance mutations by using a hybridization technique with specific probes and by comparing the results with those of the direct sequencing technique. To evaluate the temporal relationship between variations in the level of HBV-DNA and detection of mutant variants. To analyze the influence of several genotypes on the pattern of mutations developed and on values of viral load and alanine aminotransferase (ALT) after their development. PATIENTS AND METHOD: In 37 patients with chronic HBV infection, HBV genotype was determined using the LiPA technique. In 10 of these patients undergoing lamivudine treatment for a mean of 19.2 months, the development of lamivudine-resistant mutations was investigated. In these 10 patients, the LiPA technique was compared with direct sequencing. During lamivudine treatment, we determined HBV-DNA by polymerase chain reaction (PCR) and ALT every 3-6 months. RESULTS: The most frequent genotypes were D (45.9%) and A (18.9%); 2 patients were genotype B while 18.9% had mixed genotypes. Sequencing showed identical results except in one mixed genotype. Mutations were found in 60% of the cases. The results of sequencing were in agreement, except in the detection of mixed populations composed of mutants and wild-type (WT). Patients with genotype A showed the pattern M204I+WT in the first 12 months and those with genotype D showed the pattern L180M+M204V with or without WT at 18 months. In 5/6 cases, an increase of > 1 log10 in HBV-DNA was observed 3-8 months before the mutation was detected by LiPA. In patients with genotype B, levels of HBV-DNA and ALT after the development of mutations was lower than basal levels and was also lower than those in patients with genotypes A and D. CONCLUSIONS: The LiPA technique for determination of HBV genotype and detection of lamivudine resistance mutations shows excellent correlation with the most complex sequencing technique. Genotype D predominates in southern Seville. During lamivudine treatment, an increase in the level of HBV-DNA detected by PCR predicts the development of mutations before these are demonstrated by LiPA. PMID- 15544737 TI - [Autoimmune hepatitis in patients with a diagnosis of multiple sclerosis]. AB - Autoimmune hepatitis (AIH) is a chronic necroinflammatory liver disorder associated with hypergammaglobulinemia and circulating autoantibodies. Two patients previously diagnosed with multiple sclerosis who developed AIH are reported. One patient showed acute presentation with fulminant hepatic failure requiring liver transplantation. Serum autoantibodies were negative in both patients but a characteristic clinical course in the first patient as well as the hepatic histological features with typical pathological changes of AIH in both patients and a score compatible with AIH established the diagnosis. PMID- 15544738 TI - [Spontaneous aneurysmal portohepatic fistula: imaging diagnosis]. AB - Intrahepatic portosystemic venous shunts not related to trauma or biopsy are infrequent and their etiology is controversial. A congenital or acquired origin due to cirrhosis and portal hypertension has been proposed. Hepatic encephalopathy is present when there is associated cirrhosis. We describe a case of aneurysmal portohepatic venous fistula that was incidentally diagnosed with conventional ultrasonography and was subsequently confirmed by Doppler ultrasonography and computed tomography scan. Because there were no symptoms of encephalopathy, no surgical or vascular percutaneous treatment was provided. PMID- 15544739 TI - [Recurrent ingestion of foreign bodies and abdominal self-injury as a form of attempted suicide]. AB - Accidental ingestion of foreign bodies is fairly common in the general population. However, recurrent, deliberate ingestion of foreign bodies associated with continual abdominal self-mutilation with blunt objects is extremely unusual. We present the case of a young woman with an underlying psychiatric disorder who repeatedly presented to the emergency department of our hospital for attempted suicide. The most frequent reason for presenting to the emergency department was ingestion of multiple objects, usually sharp metal objects, frequently associated with abdominal self-mutilation using a sharp instrument. Treatment consisted of extraction of the foreign bodies through upper gastrointestinal endoscopy or abdominal surgery. The patient has survived all these attempts with minimal morbidity. PMID- 15544740 TI - [Consensus document on the treatment of ascites, dilutional hyponatremia and hepatorenal syndrome in liver cirrhosis]. PMID- 15544741 TI - [Primary sclerosing cholangitis]. PMID- 15544742 TI - [Insulin resistance. A cause or consequence of liver disease?]. PMID- 15544743 TI - [Octreotide and angiodysplasia]. PMID- 15544744 TI - [Elevated CA 125 in chronic liver disease with ascites]. PMID- 15544745 TI - [Acute hepatitis associated with Camellia thea and Orthosiphon stamineus ingestion]. PMID- 15544746 TI - [Gastric trichobezoar complicated with perforated gastric ulcer]. PMID- 15544747 TI - [From efficacy to effectiveness in the secondary prevention of coronary heart disease]. PMID- 15544748 TI - [Percutaneous coronary intervention for left main coronary artery disease: is it time to change the guidelines?]. PMID- 15544749 TI - [Temporary transvenous pacing: a simple and safe technique?]. PMID- 15544750 TI - [Effect of a simple educational program for physicians on adherence to secondary prevention measures after discharge following acute coronary syndrome. The CAM Project]. AB - INTRODUCTION AND OBJECTIVES: Adherence to established guidelines for patients discharged from the hospital after acute coronary syndrome is known to be suboptimal. The aim of this study was to assess the efficacy of a program for physicians centered on the treatment of acute coronary syndrome. PATIENTS AND METHOD: 39 hospitals participated. INTERVENTION: a set of measures was developed by consensus for the creation and distribution of educational materials. OUTCOMES OF INTEREST: Proportion of patients in whom ejection fraction and residual ischemia were evaluated, treatment at discharge, and health and dietary recommendations to patients (smoking, diet, exercise, etc.) referred to all patients in whom these measures or treatments should have been used ("ideal patients"). Changes were assessed with four cross-sectional surveys. RESULTS: A total of 1157, 1162, 1149 and 1158 patients were included. There were no relevant differences between these groups in baseline characteristics. In general, there was improvement in all variables between the first and the last survey. The proportion of patients who were weighed and measured increased (from 33.5% to 53.4%; P<.0001), as did the proportion of those in whom cholesterol was measured early (42.6 to 53.7%; P=.006). The proportion in whom residual ischemia was not measured despite indications for this test decreased (18.2% to 10.8%; P=.013), and the proportion increased for appropriate treatment with statins on discharge (68.6% to 81.4%; P<.0001), advice to quit smoking (60.1% to 72.2%; P<.0001) and advice to exercise (58.3% to 67.4%; P=.003). CONCLUSIONS: The educational intervention seems to have had a positive effect on improving the appropriateness of procedures and treatments for patients discharged after acute coronary syndrome. PMID- 15544751 TI - [Immediate and long-term outcome after angioplasty with stenting of the left main coronary artery]. AB - INTRODUCTION AND OBJECTIVES: Coronary artery bypass graft surgery is the treatment of choice for severe left main coronary artery stenosis. The results of a number of multicenter trials have suggested angioplasty with stenting as a possible alternative treatment. The aim of the present study was to analyze the immediate and long-term results of angioplasty with stenting of the left main coronary artery, and to identify factors predictive of death. PATIENTS AND METHOD: A total of 38 nonconsecutive patients (mean age 69 [8] years) with a severe lesion in the left main coronary artery were treated with angioplasty and stenting between November 1997 and March 2003. The procedure was elective in 27 patients and urgent in the remaining 11. In 23 patients (60.5%) the left main coronary artery was not protected by aortocoronary bypass. All patients underwent clinical follow-up examination at 25 (20) months. RESULTS: Angiographically documented success was obtained in all patients. However, one patient died from acute occlusion one hour after the operation. Four patients (10%) had a non-Q wave myocardial infarction. In-hospital mortality was 15.8% (6/38 patients). Five of the 11 patients (45.4%) who underwent emergency angioplasty and stenting died in the hospital from acute myocardial infarction complicated by severe (Killip grade III-IV) heart failure. However, only one of 27 patients (3.7%) in the elective surgery group died (P=.007). Major clinical cardiac events during follow up occurred in 5 patients (13%); 3 died and the other 2 had recurrent angina. All patients who died had an unprotected left main coronary artery. Cumulative survival rates for the elective group were 92 (0.5)% at 6 months, 88 (0.6)% at 1 year and 86 (0.7)% at 3 years, respectively. For the emergency surgery group cumulative survival rate was 54 (0.2)% at 6 months (P<.05). CONCLUSIONS: Elective angioplasty and stenting of the left main coronary artery in selected patients was associated with a high immediate success rate. In patients who underwent elective angioplasty and stenting, the incidence of major cardiac events during follow-up was relatively low. Emergency angioplasty and signs of left ventricular dysfunction were the main predictors of in-hospital mortality. PMID- 15544752 TI - [Short- and medium-term outcomes of percutaneous coronary intervention for unprotected left main coronary artery disease in patients who are poor candidates for surgical revascularization]. AB - INTRODUCTION AND OBJECTIVES: Surgical revascularization is the procedure of choice for unprotected left main coronary artery stenosis, but it may be unsuitable in some patients. We report short- and medium-term outcomes of percutaneous coronary intervention for unprotected left main coronary artery stenosis in a series of patients who were poor candidates for surgery. PATIENTS AND METHOD: Descriptive study of a historic cohort of consecutive patients with unprotected left main coronary artery stenosis who were not candidates for surgery, treated with percutaneous coronary intervention at a single center between April 1999 and December 2003. RESULTS: A total of 83 patients (mean age 72 [9] years) were included. Twenty patients (24%) were in shock on presentation. Surgery was considered unsuitable because of unacceptable surgical risk, poor condition of the distal vessels or comorbid conditions in 61 (73.5%) patients, or acute myocardial infarction in 22 (27%). An intraaortic balloon pump was used in 34 (40%); abciximab in 30 (36%) and stenting in 79 (95%) procedures. The intervention was considered successful in 76 patients (92%). Total in-hospital mortality was 28.9% (55% in patients with acute myocardial infarction and 20% in those without acute myocardial infarction). Median follow-up was 17 months. Average survival was 19.7 (2) months. Eighteen (22%) patients were hospitalized again for a new ischemic event, and 14 (17%) underwent revascularization. In 9 cases (10.8%) a new angioplasty was performed, and in 5 (6.0%) surgical revascularization was necessary. CONCLUSIONS: Percutaneous coronary intervention is an option for revascularization in left main coronary artery stenosis in patients who are poor candidates for surgery, although in-hospital and long-term mortality remain high. PMID- 15544753 TI - [Temporary pacemakers: current use and complications]. AB - INTRODUCTION AND OBJECTIVE: Temporary pacemakers (TP) are used in the emergency treatment of patients with severe bradyarrhythmia. They are often used in emergency situations and for older patients in poor general condition who are hemodynamically unstable and uncooperative. The aim of this study was to review and analyze the indications, incidence and type of complications associated with TP implanted in our center during a 6-year period. PATIENTS AND METHOD: We analyzed significant clinical variables, indication, route of insertion, follow up, complications, and duration of temporary pacing. RESULTS: A total of 568 TP were implanted, and 530 cases were available for review (mean age 74.8 [11] years). The main indications were symptomatic complete AV block (51%), prophylaxis for replacement with a definitive pacemaker (14.7%), blockage in the acute phase of myocardial infarction (12.6%), bradyarrhythmia due to drug intoxication (12.2%), symptomatic sick sinus syndrome (7.5%) and long QT interval or ventricular tachycardia (2.5%). The route of insertion was via the femoral vein in 99% of the cases. The duration of TP use was 4.2 days (range 1 to 31 days). A total of 369 patients (69.6%) required a permanent pacemaker. COMPLICATIONS: 34 patients died (6.4%), but only 3 deaths were attributable to TP implantation. Other severe complications were seen in 98 patients (18.5%). Malfunction of the TP occurred in 48 patients (9%) because of electrode displacement. CONCLUSIONS: Temporary pacemakers are used in older patients with extreme bradyarrhythmia and occasionally with acute myocardial infarction. Serious complications are not uncommon (22% of all patients), and can range from femoral hematoma to cardiac tamponade and even death (6%). In 9% of the patients the electrode needed to be repositioned because of failure of sensing or loss of ventricular capture. PMID- 15544755 TI - [Assessment of left atrial wall velocities by pulsed wave tissue Doppler imaging. A new approach to the study of atrial function]. AB - INTRODUCTION AND OBJECTIVES: To analyze the profile of left atrial wall velocities by pulsed wave tissue Doppler imaging, and to compare the relationship between these observations and the transmitral and pulmonary vein flow velocities obtained by conventional pulsed Doppler echocardiography. PATIENTS AND METHOD: We studied 90 patients (50 women and 40 men, mean age 48 [22] years). Pulsed tissue Doppler images of the left atrial wall were obtained and analyzed in all subjects. The study population was then divided in two groups: group I (age < 45 years) and group II (age > 45 years). Transmitral and pulmonary vein flow velocity tracings were obtained simultaneously by pulsed Doppler echocardiography. RESULTS: With pulsed tissue Doppler interrogation of the left atrial wall, a triphasic signal was recorded in all patients, consisting of a positive wave (A1) followed by two negative waves (A2 and A3). Younger subjects (group I) showed a pattern with a prominent A2 wave and an A2/A3 ratio > 1. In older patients (group II) peak velocity of the A2 wave diminished and peak velocity of the A3 wave increased, so that the A2/A3 ratio was < 1. We found no differences in peak velocity of the A1 wave between the two age groups (13.5 (3.9) cm/s in group I vs 13.1 (5.4) cm/s in group II; P = .59). Significant concordance was observed between the transmitral flow pattern and the left atrial pulsed tissue Doppler tracing (kappa = 0.584; P < .0001). CONCLUSIONS: Evaluation of the left atrial wall using pulsed tissue Doppler imaging is feasible and reproducible. Tissue Doppler imaging provides new quantitative insights of potential use in the assessment of left atrial function. PMID- 15544754 TI - [Effects of carvedilol on functional capacity, left ventricular function, catecholamines and oxidative stress in patients with chronic heart failure]. AB - INTRODUCTION AND OBJECTIVE: Carvedilol is an antioxidant and adrenergic antagonist with demonstrated benefits in terms of mortality from heart failure (HF). The aim of the present study was to evaluate clinical parameters, left ventricular function, oxidative stress levels and neurohumoral status at baseline and after 6 months of treatment with carvedilol in patients with chronic HF. PATIENTS AND METHOD: Thirty patients with chronic HF that was stable without beta blocker treatment were included. Functional class was II or III, and left ventricular ejection fraction (LVEF) was < 40%. Mahler score, distance walked in 6 min, peak oxygen consumption, LVEF, plasma catecholamine (norepinephrine) concentration and oxidative stress parameters were evaluated at baseline and after 6 months of treatment with carvedilol. RESULTS: Mean age was 59 (2) years, and 23 patients were men. After 6 months of treatment there were clinical improvements as measured by the Mahler score (from 6.8 to 11.0 points; P=.001) and the 6-min walk distance (from 499 [18] to 534 [17] m; P =.032), but no changes in peak oxygen consumption. The LVEF increased from 24 (1) to 31 (2)% (P=.003). In patients with chronic HF, plasma malondialdehyde concentration was significantly lower after 6 months (decrease from 2.4 [0.2] to 1.1 [0.2] micromol/l; P<.001). No significant changes were observed in plasma catecholamine levels or antioxidant enzyme activities. CONCLUSIONS: In patients with chronic HF, carvedilol treatment for 6 months was associated with clinical improvements, increased left ventricular function and decreased plasma concentrations of malondialdehyde, with no changes in plasma catecholamine levels. PMID- 15544756 TI - [Spanish Registry of Catheter Ablation. Third Official Report of the Working Group on Electrophysiology and Arrhythmias of the Spanish Society of Cardiology (2003)]. AB - INTRODUCTION: The results of the Third Spanish Catheter Ablation Registry, developed by the Working Group on Electrophysiology and Arrhythmias, are presented for the third consecutive year (2003). MATERIAL AND METHOD: In contrast to previous years, data were collected in two different ways at the discretion of the participating center. Retrospective were obtained with a standard questionnaire, as in previous years. Prospective data were obtained from a database of records completed after each ablation was performed. Results and complications are presented according to different arrhythmic substrates. RESULTS: Thirty-nine centers participated in the registry (25 supplied prospective data and 14 retrospective data), representing more than 80% of all electrophysiological laboratories in Spain. A total number of 4354 ablations were recorded (111 procedures per center, 2723 from the prospective registry and 1631 from the retrospective one). The substrate most frequently treated was AV nodal reentry tachycardia (31.6%, 98% success), followed by accessory pathways (26%, 89% success) and atrial macro-reentry tachycardia (23%, 90% success). The incidence of complications was 1.7% and mortality was 0.11%. With the prospective registry we obtained more comprehensive information individualized for each procedure (age, sex, underlying cardiomyopathy, anticoagulation, sedation, type of catheter, etc.). CONCLUSIONS: The high rate of participation in the registry and the consistency of the results with previous years help to consolidate the registry as a reference for the rest of the scientific community. The results from the prospective registry showed better-quality information and more detailed reporting of results and complications. PMID- 15544757 TI - [Spanish Registry of Cardiac Catheterization and Coronary Interventions. Thirteenth Official Report of the Working Group on Cardiac Catheterization and Interventional Cardiology of the Spanish Society of Cardiology (1990-2003)]. AB - The results of the Registry of the Working Group on Cardiac Catheterization and Interventional Cardiology of the Spanish Society of Cardiology for 2003 are presented. Data were obtained from 112 centers representing nearly all cardiac catheterization laboratories in Spain; 104 centers performed mainly adult catheterization and 8 carried out pediatric procedures only. In 2003, 105,939 diagnostic catheterization procedures were performed, including 90 939 coronary angiograms, representing a total increase of 8.5% in comparison to 2002. The population-adjusted rate was 2171 coronary angiograms per 106 inhabitants. Coronary interventions increased by 14.4% in comparison to 2002, with a total of 40,584 procedures and a rate of coronary interventions of 969 per 106 inhabitants. Coronary stents were used in 92.5% of the procedures (47,249 units implanted, for a total increase of 22% in comparison to 2002). About one fifth (20.2%) of the implanted stents were drug-eluting stents (11,699 units). A total of 6080 percutaneous coronary interventions were done in patients with acute myocardial infarction, representing an increase of 27.5% in comparison to 2002, and accounting for 14.9% of all interventional procedures. Of the noncoronary interventions recorded, we note the increase in percutaneous mitral valvuloplasties (21.6%) and atrial septal defect closures (86%), and also the increase in pediatric interventions (13.3%). In conclusion, we emphasize the high rate of reporting by laboratories, which allows the Registry to compile data that are highly representative of the activity at cardiac catheterization laboratories in Spain. PMID- 15544758 TI - [Guidelines on the diagnosis and management of pericardial diseases. Executive summary]. PMID- 15544759 TI - [Pulmonary thromboembolism and multislice tomography]. PMID- 15544760 TI - [Myocardial perfusion SPECT in right bundle branch block and left anterior hemiblock]. AB - The aim of this study was to determine the diagnostic accuracy of myocardial perfusion SPECT (single photon emission computed tomography) with technetium labeling in patients with right bundle branch block (RBBB) and with left anterior hemiblock (LAH). One hundred seven patients (53 RBBB, 54 LAH) for whom coronary angiography results were available were selected. The gold standard for diagnosis was based on two coronary angiography criteria: a) stenosis equal to or greater than 50% in at least one coronary artery, and b) completely normal coronary angiography. The sensitivity of SPECT was approximately 90% in both RBBB and LAH. Specificity was 58% and 50%, respectively, for the first criterion, but was 85.7% and 87.5%, respectively, for the second criterion. PMID- 15544761 TI - [Budd-Chiari syndrome with complete occlusion of the inferior vena cava: percutaneous recanalization by angioplasty and stenting]. AB - A 47-year-old man was diagnosed with primary antiphospholipid syndrome and Budd Chiari syndrome (membranous complete obstruction of the intrahepatic inferior vena cava), with edema and ascites refractory to medical treatment. The inferior vena cava membrane was punctured with a Brockenbrough needle under multidirectional fluoroscopic guidance via a transfemoral approach. The occlusion was dilated with balloons of increasing size and was subsequently stented successfully. At 1-year follow-up venography showed patency of the stent, and the patient remains asymptomatic 2 years after the procedure. PMID- 15544762 TI - [Transient ventricular dysfunction after emotional stress]. AB - We describe a patient who developed severe ventricular dysfunction and cardiogenic shock after intense emotional stress. Her subsequent course was favorable, with complete recovery of left ventricular systolic function. The coronary arteries were normal and no specific etiologic agent was demonstrated. Her clinical picture was compatible with transient left ventricular apical ballooning. [I123]metaiodobenzyl guanidine cardiac scintigraphy showed a marked decrease in cardiac sympathetic nerve activity. We discuss the pathophysiologic mechanisms of this syndrome. PMID- 15544764 TI - [Can we expect polymorphisms to answer our questions?]. PMID- 15544765 TI - Ludo ergo sum. PMID- 15544766 TI - Classification of congenital melanocytic naevi and malignant transformation: a review of the literature. AB - INTRODUCTION: Congenital naevi (CN) vary greatly in size, macroscopic appearance and histology. There is a practical need to subdivide CN according to size, since size differences have a direct bearing on cosmetic and resultant psychological problems, and on therapeutic options, and probably on the chance of malignant transformation. In this review, we summarise the literature on size subgroupings of CN, with special focus on giant congenital naevi and their risk of malignant transformation. MATERIALS AND METHODS: A Medline literature search from 1966 to October 2002 was performed. Only English-language studies focusing on CN in association with melanoma were included. The final strategy consisted of textwords and medical subject heading (MeSH) terms on small, medium, large and giant congenital naevi combined with the textwords classification, histology and melanoma. Additional manual cross-referencing was performed. We excluded articles that dealt only with aspects of treatments. RESULTS: A wide variety of criteria for size subgrouping of CN has been put forward in the literature and precludes a direct comparison of reported data (Table 1). We identified 35 such articles in the world literature in which no less than seven different definitions of minimum size of a giant CN were employed. Histologically, it is difficult or even impossible to conclude that a naevus is congenital or acquired, especially in case of a small lesion, since the differences are not absolute (Table 2). Giant CN have an increased risk for malignant transformation, but the reported incidence rates have differed widely from one to 31% (Table 3). Reported melanoma incidence rates have derived from retro- and prospective studies, reviews and case reports, and compared with each other using different definitions. On top of this, patients in different age groups were reported, who were registered in different referral centers. CONCLUSION: To allow comparison of study results from different centers, it is essential that the size subclassification of CN is based on standard and generally accepted criteria. We recommend defining GCN as a CN covering one percent body surface area in face and neck and two percent elsewhere on the body. Based on a review of the world literature, we recommend prophylactic excision of all CN, in close communication with patient and family and individualising treatment accordingly. PMID- 15544767 TI - Combined early treatment of congenital melanocytic naevus with carbon dioxide and NdYag lasers. AB - Three cases of extensive congenital melanocytic naevi were treated with a combined approach using a ultrapulse carbon dioxide laser and a Nd Yag laser within 1 year of birth. The naevi were substantially depigmented and this result has been maintained for up to 36 months following treatment. The condition, treatment and the relevant literature are discussed. PMID- 15544768 TI - Contraindications to sentinel lymph node biopsy in cutaneous malignant melanoma. AB - This paper discusses possible contraindications to sentinel node biopsy in cutaneous malignant melanoma based on a large series from our Melanoma Unit. PMID- 15544769 TI - The Waist Coat Flap: a new technique for closure of infected median sternotomy wounds. AB - The ideal standard of treatment of the infected sternotomy wound is early recognition, adequate debridement and repair with well vascularised tissue. We describe a new technique The Waist Coat Flap, which adheres to these principles, but does not require muscle or omentum and does not compromise their future use, if required. PMID- 15544770 TI - Ten year experience of bioabsorbable mesh support in pectus excavatum repair. AB - INTRODUCTION: We reviewed 10 years experience in the treatment of this deformity using a retro-sternal bioabsorbable mesh in place of a metallic device to lift and stabilise the sternum. Moreover, the mesh supports the thoracic and upper abdominal wall reconstruction. MATERIALS AND METHODS: From January 1990 to December 2000, in our Thoracic Surgery Unit, 65 patients with PE were assessed for surgical repair, mean age 16+/- 3.5 years, fronto sagittal thoracic index (FSTI) 0.21, ranging from 0.15-0.33. Twenty-three of them underwent surgical correction after initial assessment, 22 were deferred and sent to physiotherapy. At a subsequent assessment, five of the patients sent to physiotherapy were deemed to require surgery. RESULTS: Of the 28 patients who underwent surgery, 2 (10%) presented a mild recurrence of PE after 1 year (0.300.34), meanwhile all other patients maintained a FSTI>0.34. For all patients the improvement in FSTI was statistically significant, p = 0.001. Patients satisfaction after 24 months was thus shared: excellent 18 patients (65%), good seven patients (25%), fair one patient (3.5%) and poor two patients (7%). No major complications were observed in preoperative period. Patients mobilisation was soon achieved thanks to the postoperative pain control and the absence of retro-sternal metallic support. CONCLUSIONS: The introduction of bioabsorbable mesh in the Robicsek technique is a safe procedure related to a high percentage of success. The high tolerance of the material reduces the inflammatory reaction. Moreover, the procedure prevents patients from having complications caused by retro-sternal device dislodgment, avoiding a second intervention for device reposition and reducing the postoperative chest pain achieving an early patient mobilisation. In the end a complete reconstruction of the upper abdomen wall has been produced. PMID- 15544771 TI - A new umbilicoplasty for children: creating a longitudinal deep umbilical depression. AB - When forming a deep umbilical depression for paediatric patients displaying a thin layer of abdominal fat, the V-Y advancement flap method is useful. However, although many plastic surgeons approve of a longitudinal umbilical depression as an ideal, conventional flap methods often result in only a longitudinal scar line with a depression at the end of the scar. The shifted umbilical depression facing upward or downward appears aesthetically unnatural. To resolve these problems, we have devised a new method of umbilicoplasty. Using obliquely opposing flaps, a longitudinal deep umbilical depression facing forward can be created at the correct umbilical position. PMID- 15544772 TI - Early reconstruction of the abdominal wall in giant omphalocele. AB - Omphalocele is the most common congenital defect of the abdominal wall. Mortality rate is between 20 and 70% and early closure of the abdominal wall, within 10 days of life, is vital to the successful outcome of the surgical treatment. The authors describe the use of two bipedicled flaps of abdominal skin to correct the defect of the midline as soon as the reduction of all viscera has been accomplished. PMID- 15544773 TI - Quality of life in patients with vascular malformations of the lower extremity. AB - With the exception of capillary malformations (port-wine stains), the adverse psychosocial effects of vascular malformations have not received much attention in the medical literature. We, therefore, studied health related quality of life of patients with vascular malformations located primarily on the lower extremity, who presented to us over a 10-year period. Patient's self-assessment of quality of life was measured by the Multiple Outcomes Study (MOS) Short Form Health Survey Questionnaire (SF-36). Several possible predictors were also examined. Eighty-one patients (33 male, 48 female), aged 14-61 years, completed the SF-36. Seventy-one patients (88%) had a low flow lesion. Twenty-three patients (28%) had hypertrophy of the lower extremity, while 11 patients (14%) had hypotrophy. Sixty nine patients (85%) had the vascular malformation located only on the lower extremity. Six (7%) also had their upper extremity (hand) involved, and 20 patients (24.7%) had >10% TBSA affected. Eight patients (10%) needed special shoes. Fifty-one patients (63%) had a previously performed MRI, of which 62.7% (32/51) had muscle involvement. Fifty-one patients (63%) had been operated on. Of the 34 patients wearing elastic compression stockings, 25 patients (74%) indicated that they were satisfied with the stockings. Compared to the general population sample, the SF-36 questionnaire indicated that our vascular malformation patients reported impaired vitality and higher levels of pain, while no differences were seen regarding the other dimensions of quality of life. Demographic, clinical and therapy characteristics could explain quality of life only to a limited extent. Explained variability of the SF-36 dimensions ranged from 0% for mental health to 34.5% for role functioning physical. To our surprise, this study suggests that patients with vascular malformations located primarily on the lower extremity do not have a greatly decreased quality of life when compared to the general Dutch population. PMID- 15544774 TI - The buccal fad pad lined with a metabolic active dermal replacement (Dermagraft) for treatment of defects of the buccal plane. AB - The buccal fad pad flap (BFPF) is an easy to raise pedicled flap for closure of intraoral defects with barely any donor defect. The major disadvantage of the BFPF is the missing epithelial lining, which can induce fibrous tissue formation with resulting functional impairment. To overcome this problem we lined the BFPF with a dermal replacement (Dermagraft) consisting of living metabolic active fibroblasts. In six patients, defects resulting from tumour removal were reconstructed with a combination of the BFPF and Dermagraft and followed up for at least 2 years. In all patients a defect closure was achieved with no functional impairment. The clinical character which was achieved was more a type of defect regeneration than a flap closure. The availability of bioengineered metabolic active tissue can overcome the major problem of the BFPF as an option for defect closure of the buccal side. Furthermore the regeneration of the defects optimises clinical tumour monitoring. PMID- 15544775 TI - The influence of sildenafil on random skin flap survival in rats: an experimental study. AB - Sildenafil (Viagra), a selective and specific inhibitor of cyclic guanosine monophosphate (cGMP) phosphodiesterases (PDEs), is currently marketed for the treatment of erectile dysfunction. Sildenafil is a potent and highly selective PDE-V inhibitor and enhances smooth muscle relaxation in human. Systemic arterial and venous smooth muscle cells contain PDE-V and nitric oxide (NO) which is a major mediator of relaxation of the smooth muscle cell. The aim of the present study is to investigate, in a rat model, the potential effect of sildenafil on survival of random pattern skin flaps. For this purpose, 32 Sprague-Dawley rats were used and a McFarlane-type caudally based skin flap was designed on the dorsum of the rat (2.5 x 8 cm). Rats were divided into four groups: One control (Group D), and three treatment groups (Groups A, B, C). Sildenafil was administered orally to the experiment groups; Group A: 3 mg/kg/single dose a day, Group B: 10 mg/kg/single dose a day and Group C: 10 mg/kg/twice dose a day. The areas of flap necrosis were measured in each group. The extent of viable flap areas were expressed as a percentage of total flap area, and differences were studied by Completely Randomised Experimental design. The areas of necrosis of skin flaps decreased depending on sildenafil dose, but viability of the flaps treated with 3 mg/kg/day was not different than the control group. The flaps receiving 2 x 10 mg/kg/day sildenafil gave the highest (P < 0.01) survival rate. As a conclusion, sildenafil may have a dose dependent effect to increase flap survival in random skin flaps. PMID- 15544776 TI - Plastic surgery in the undergraduate curriculum: the importance of considering students' perceptions. AB - As the undergraduate medical curriculum becomes increasingly crowded the competition for time inevitably increases and surgical specialties have decreasing representation. Plastic surgery is regarded with some confusion in terms of its relevance to the generic doctor. Plastic surgeons have no doubt about the relevance of the specialty to undergraduates. Others see this as a very technical specialty dealing with complex reconstructions and surgical interventions or, as a rather indulgent specialty focusing mainly on glamour and cosmesis. This study focuses on students' perceptions of an undergraduate teaching program in plastic surgery. The reality is that highly pressured undergraduates do not have the luxury of time to consider the finer details of the specialties to which they are exposed. Their priority is to pass their examinations and, having addressed that concern, further information becomes an acceptable bonus. The conclusion is that if plastic surgeons are going to gain greater involvement in the undergraduate curriculum they must start with involvement in examinations and assessments. The students will then ensure that adequate and appropriate teaching time is allocated. PMID- 15544777 TI - Vascular supply of the subcutaneous pedicle of Nagata's method in microtia reconstruction. AB - Nagata's method is a two-stage method for total ear reconstruction in patients with microtia. In the first stage of this procedure, mastoid flap and posterior lobule flap are elevated with a subcutaneous pedicle. However, contribution to the vascular supply by this pedicle has been controversial. We investigated the presence or absence of apparent vessels in the subcutaneous pedicle in 14 primary cases of microtia in the first stage operation. In all cases some vessels were included in the pedicle. In lobular and small concha type microtia, the vessels originated from the parotid fascia or aponeurotic tissue behind the remnant cartilage. In concha type microtia, apparent vessels could be preserved by including the perichondrium of the conchal cartilage. These findings suggest that the mastoid and posterior lobule flaps or W-shaped flap in Nagata's first stage operation are actually the perforator-based flaps. The source vessel of the perforators seemed to be the posterior auricular artery because of its location although further dissection was not performed in order not to damage the vascular supply. The presence of the vessels can augment the blood supply of not only W shaped flaps but also the skin flap cephalad to them. By confirming the preservation of the perforators in the subcutaneous pedicle the surgeon may be able to trim the covering skin more safely. PMID- 15544778 TI - Early presentation of an extremity arteriovenous malformation. AB - We report a very rare case of a high flow arteriovenous malformation (AVM) of the upper limb that caused high output cardiac failure at birth. There was early transfer of the baby to the care of a multidisciplinary team. After radiological intervention, the arm distal to the malformation became ischaemic and an urgent amputation through the upper-humerus followed. Methods of treatment are discussed, together with a review of results in the literature. PMID- 15544779 TI - Chest hair micrografts display extended growth in scalp tissue: a case report. AB - Conventional follicle harvesting techniques for hair transplantation are limited by the available scalp donor hair. The development of an innovative technique of microsurgical single follicular unit extraction has made it possible to exploit body hair grafts for scalp transplantation. This case study reports on 18 months of follow-up on a patient with extensive scalp scarring who underwent a transplantation procedure using donor chest hair. The photographically documented results show a change in the length of the chest hair measuring an average of 4 cm at transplant to 15 cm by 18 months post-transplant. The transplanted chest grafts provided an excellent cosmetic result for hair replacement. PMID- 15544780 TI - Congenital cleft ear: a case report. AB - A case is presented of congenital incomplete clefting of a normally formed ear. Previous reports have been limited to clefting of the ear lobule or clefting in association with gross deformity of the ear. PMID- 15544781 TI - Correction of congenital 'triple lobe type' auricular cleft. AB - Congenital cleft earlobe is a rare auricular malformation. Various techniques have been reported for the correction of this deformity. Few papers deal with surgical techniques to correct the triple lobe sub-type cleft. This anomaly presents more technical difficulties in the three-dimensional rearrangement than other sub-types. We describe the technique used to correct a triple lobe deformity in a 5-year-old boy. The postoperative result was satisfactory at one year follow-up. PMID- 15544782 TI - Improved selection and management of the auricular composite graft donor site. PMID- 15544783 TI - Supernumerary auricle on the lateral canthus. AB - A boy was born with an appendage on his right lateral canthus, with associated supernumerary auricles on the right cheek and a right ocular dermoid. We resected the appendage. Its core was composed of elastic cartilage, as is the external auricle. The lateral canthus overlaps facial cleft line No. 8 in Tessier's classification [Plast Reconstr Surg 4 (1976) 69] and forms the upper part of the first branchial arch. It appears that our patient's appendage was a supernumerary auricle, which had developed from the first branchial arch. PMID- 15544784 TI - The unilateral pectoralis major island flap, an efficient and straightforward procedure for reconstruction of full length sternal defects after postoperative mediastinal wound infection. PMID- 15544785 TI - The treatment of naevus comedonicus. PMID- 15544786 TI - Erosive pustular dermatosis of the scalp occurring after partial thickness skin graft for squamous cell carcinoma. PMID- 15544787 TI - Use of methylene blue in the complete excision of ruptured cutaneous cysts. PMID- 15544788 TI - Synthesis, central and peripheral benzodiazepine receptor affinity of pyrazole and pyrazole-containing polycyclic derivatives. AB - A series of new pyrazole-condensed 6,5,5 tricyclic compounds were synthesized and tested to evaluate their binding affinities at both central (CBR) and peripheral (PBR) benzodiazepine receptors. Some 1-aryl-5-phenylpyrazole derivatives were also prepared and tested for comparison with their corresponding rigid tricyclic analogs. Among the newly synthesized 1-aryl-1,4-dihydro-indeno[1,2-c]pyrazoles bearing both an ethoxycarbonyl group at position 3 and a carbonyl function at the position 4, compound 4b emerged as a new potent (IC(50) = 26.4 nM) and selective CBR ligand. The 4-oxo-1-aryl-1,4-dihydro-indeno[1,2-c]pyrazole diethylamide derivative 14a was instead identified as a relatively potent (IC(50) = 124 nM) but highly selective PBR ligand. PMID- 15544789 TI - Stable propylphosphonic acid analogues of geranylgeranyl diphosphate possessing inhibitory activity on geranylgeranyl protein transferase. AB - Stable analogues of geranylgeranyl diphosphate, possessing 3 (phosphono)propionamido moieties in the place of the metabolically unstable diphosphate portion, were prepared and submitted to prenyltransferase (GGTase and FTase) inhibition assays. In one case, an excellent GGTase inhibitory activity was obtained (IC(50) = 39 nM), accompanied by a certain degree of GGTase vs. FTase selectivity. PMID- 15544790 TI - Azaanalogues of ebselen as antimicrobial and antiviral agents: synthesis and properties. AB - The different analogues of ebselen-unsubstituted benzisoselenazol-3(2H)-one (2a) 2-pyridylbenzisoselenazol-3(2H)-ones (2b-h) and 7-azabenzisoselenazol-3(2H)-ones (3a-j) were designed as new selenium-containing antiviral and antimicrobial agents and synthesized. Some of them were found in the antiviral assay in vitro to be strong inhibitors of cythopatic activity of herpes simplex virus type 1- HSV-1 (compounds 2a,b,f,h, 3a-j) and encephalomyocarditis virus--EMCV (compounds 2a,h, 3a-f,k,l). The compounds 2a,h and 3a-e,j were found to have an appreciable activity against Gram-positive bacteria (Staphylococcus aureus and Bacillus) in vitro, some of them inhibited growth of pathogenic yeasts (Candida albicans) (3a,b) and filamentous fungi (3a-e,f). PMID- 15544791 TI - Synthesis, characterization and transfection activity of new saturated and unsaturated cationic lipids. AB - We synthesized new cationic lipids, analogue to N-[1-(2,3-dioleoyloxy)propyl] N,N,N-trimethylammonium chloride (DOTMA) and 1,2-dimyristyloxypropyl-3-dimethyl hydroxyethylammonium bromide (DMRIE), in order to compare those containing a dodecyl chain with those having a relatively long chain with two or five double bonds, such as squalenyl and dihydrofarnesyl derivatives, or complex saturated structures, such as squalane derivatives. The fusogenic helper lipid dioleoylphosphatidylethanolamine (DOPE) was added to cationic lipids to form a stable complex. Liposomes composed of 50:50 w/w cationic lipid/DOPE were prepared and incubated with plasmidic DNA at various charge ratios and the diameter and zeta potential of the complexes were measured. The surface charge of the DNA/lipid complexes can be controlled by adjusting the cationic lipid/DNA ratio. Finally, we tested the in vitro transfection efficiency of the cationic lipid/DNA complexes using different cell lines. The transfection efficiency was highest for the dodecyloxy derivative containing a single hydroxyethyl group in the head, followed by the dodecyloxy and the farnesyloxy trimethylammonium derivatives. Instead the C27 squalenyl and C27 squalanyl derivatives resulted inactive. PMID- 15544792 TI - Syntheses, in vitro antibacterial and cytotoxic activities of a series of 3 substituted succinimides. AB - We have synthesized a series of 3-substituted succinimides and their in vitro antibacterial activities have been tested towards Gram-positive and Gram-negative bacteria from the ATCC collection. Some of them possess significant antibacterial activity against Gram-positive organisms (Staphylococcus aureus ATCC 25923 and Enterococcus faecalis ATCC 29212) but all are poorly active or inactive against Gram-negative organisms (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853). The compounds with the lowest minimal inhibitory concentrations (esters of 3-hydroxy succinimides) are also the most cytotoxic against green monkey Vero cell line (ATCC CCL-81) and could explain that perhaps apoptosis should be implicated in eukaryotic cell cytotoxicity of succinimides. PMID- 15544793 TI - Phosphonomethylphosphorylmethyl(oxy)-analogues of geranylgeranyl diphosphate as stable and selective geranylgeranyl protein transferase inhibitors. AB - The diphosphate moiety of geranylgeranyldiphosphate (GGdP) was replaced with metabolically and hydrolytically stable analogous polar portions, in an attempt to obtain new geranylgeranyltransferase (GGTase) inhibitors, which could also be selective over congener enzyme farnesyltransferase (FTase). In particular, the phosphonomethylphosphorylmethoxy derivative showed the highest inhibition potency, accompanied by a satisfactory GGTase/FTase selectivity. PMID- 15544794 TI - Synthesis of some 3-(arylalkylthio)-4-alkyl/aryl-5-(4-aminophenyl)-4H-1,2,4 triazole derivatives and their anticonvulsant activity. AB - A series of novel 3-[[(substituted phenyl)methyl]thio]-4-alkyl/aryl-5-(4 aminophenyl)-4H-1,2,4-triazoles 11-20 and several related Schiff's bases, 3 [[(substituted phenyl)-methyl]thio]-4-alkyl/aryl-5-[[[(substituted phenyl/5-nitro 2-furyl)methylene]amino]-phenyl]-4H-1,2,4-triazoles 21-31 were synthesized for evaluation of their biological properties. Structures of the synthesized compounds were confirmed by the use of their spectral data besides elemental analysis. All compounds were evaluated for their anticonvulsant activity by maximal electroshock (MES), subcutaneous pentylenetetrazole (scPTZ) and neurotoxicity (NT) screens. A number of triazole derivatives, exhibited protection after intraperitoneal administration at the dose of 100 and 300 mg/kg in one or both models employed. Compounds 12, 13 and 14 were subjected to oral MES screening in rats at 30 mg/kg and were observed to protect 50% of the animals employed in the experiment. Antimicrobial and antituberculosis activity of these compounds 11-31 were also screened. Some of the tested compounds showed marginal activity against M. tuberculosis H37 Rv. PMID- 15544795 TI - The role of various surfactants on the release of salbutamol from suppositories. AB - Salbutamol is a selective beta(2)-adrenoreceptor agonist with different pharmacological effects. In this research because of the simplicity of suppository application in elderly and its higher plasma concentration than tablets as well as its particular indication in premature labour, salbutamol suppositories were prepared. The suppositories were formulated containing 10 mg of the drug and Witepsol H15, the oleaginous soluble base using melting method. To optimize the release rate of drug, different surfactants namely, sodium lauryl sulphate (SLS) as an ionic surfactant and Tween 80 as well as Arlacel 60 as non ionic surfactants with different HLBs were chosen. The effect of surfactant concentration on the release rate of salbutamol from suppositories were also investigated. All prepared formulations fulfilled the specifications set down in British Pharmacopoeia. The results showed that Tween 80 (2%w/w) and SLS (0.75%w/w) caused an increase in dissolution rate of salbutamol from suppositories. As anionic surfactants, such as SLS, cause greater damage on mucosa than non-ionic surfactant, such as Tween 80, this study recommended that Tween 80 could be added in suppository formulation in order to increase the dissolution rate of salbutamol. It was also shown that the release rate of salbutamol altered linearly with the amount of Tween 80 in suppository formulations. PMID- 15544796 TI - Spectrophotometric determination of clozapine based on its oxidation with bromate in a micellar medium. AB - A simple and sensitive spectrophotometric method was developed for the determination of clozapine in its dosage forms. The method is based on the reaction of the drug with potassium bromate in a perchloric acid medium to produce an intense yellow colored species exhibiting a maximum absorption at 308 nm. Beer's law is obeyed for up to 12.0 microg ml(-1) with a correlation coefficient (n = 6) of 0.9998 and a detection limit (3S(b)) of 0.1 microg ml(-1). The molar absorptivity is 1.986 x 10(4) l mol(-1) cm(-1). The various experimental parameters affecting the development and stability of the colored oxidation product were carefully studied and optimized. The proposed method was successfully applied to the determination of clozapine in its dosage forms. The results obtained were in good agreement with those obtained using the B.P. official method. PMID- 15544797 TI - Application of oxidants to the spectrophotometric determination of amlodipine besylate in pharmaceutical formulations. AB - Three new spectrophotometric methods for the determination of amlodipine besylate have been proposed. The first two methods, i.e. A and B, are based on the oxidation of the drug with Fe(III) and the estimation of Fe(II) produced after chelation with either 1,10-phenanthroline or 2,2'-bipyridyl at 500 and 515 nm, respectively. The Beer's law was obeyed in the concentration ranges of 2-10 and 4 14 microg ml(-1) with molar absorptivity of 2.9 x 10(4) and 2.7 x 10(4) l mol(-1) cm(-1) for methods A and B, respectively. The third procedure depends on the interaction of amlodipine besylate with ammonium heptamolybdate tetrahydrate, which resulted in the formation of molybdenum blue (lambda(max) 825 nm). The linear dynamic range and the molar absorptivity values were found to be 15-59 microg ml(-1) and 1.8 x 10(4) l mol(-1) cm(-1), respectively. The results of the proposed procedures were validated statistically and compared with those obtained by the reference method. The proposed methods were applied successfully to the determination of amlodipine besylate in commercial tablets. PMID- 15544798 TI - Optimization and statistical evaluation of dissolution tests for indinavir sulfate capsules. AB - An optimization, statistically based on t-student test, to set up dissolution test conditions for indinavir sulfate capsules is presented. Three dissolution media, including that reported in United States Pharmacopeial Forum, and two apparatus, paddle and basket, were applied. Two different indinavir sulfate capsules, products A and B, were evaluated. For a reliable statistical analysis eighteen capsules were assayed in each condition based on the combination of dissolution medium and apparatus. All tested media were statistically equivalent (P > 0.05) for both drug products when paddle apparatus was employed at the stirring speed of 50 rpm. The use of basket apparatus at the stirring speed of 50 rpm caused significant decrease in the drug release percent for the product B (P < 0.05). The best dissolution conditions tested, for products A and B, were applied to evaluate capsules dissolution profiles. Twelve dosage units were assayed and dissolution efficiency concept was used, for each condition, to obtain results with statistical significance (P > 0.05). Optimal conditions to carry out the dissolution test were 900 ml of 0.1 M hydrochloric acid as dissolution medium, basket at 100 rpm stirring speed and 260 nm ultraviolet detection. PMID- 15544799 TI - Crystal structure of the spindle assembly checkpoint protein Bub3. AB - Bub3 is one of at least six proteins that transmit the spindle assembly checkpoint signal. These proteins delay cell cycle progression from metaphase to anaphase in response to attachment defects between kinetochores and spindle microtubules and to tension defects between sister chromatids. To explore the molecular interactions mediated by Bub3, we have determined the crystal structure of the Saccharomyces cerevisiae protein Bub3p at 2.35 A resolution. Bub3p is a seven-blade beta-propeller, although its sequence diverges from that of other WD40 family members. Several loops are substantially elongated, but extra domains or insertions are not present at the termini. In particular, two extended loops project from the top face of the propeller, forming a cleft. Amino acid residues across the top face and one aspect of the lateral surface (spanning blades 5-6) are highly conserved among Bub3 proteins. We propose that these conserved surfaces are the loci for key interactions with conserved motifs in spindle checkpoint proteins Bub1 and Mad3/BubR1. Comparison of the Bub3 sequence to the WD40 protein, Rae1, shows high sequence conservation along the same surfaces. Rae1 interaction with Bub1 is, therefore, likely to involve a similar mode of binding. PMID- 15544800 TI - Regulation of transcriptional silencing in yeast by growth temperature. AB - Increasing evidence indicates that transcriptionally silent chromatin structure is dynamic and may change its conformation in response to external or internal stimuli. We show that growth temperature affects all three forms of transcriptional silencing in Saccharomyces cerevisiae. In general, increasing the temperature within the range of 23-37 degrees C strengthens HM and telomeric silencing but reduces rDNA silencing. High temperature (37 degrees C) can suppress the silencing defects of histone H4 mutants. We demonstrate that DNA at the silent HML locus becomes more and more negatively supercoiled as temperature increases in a Sir-dependent manner, which is indicative of enhanced silent chromatin. This enhancement of silent chromatin is not dependent on silencers and therefore does not require de novo assembly of silent chromatin. We also present evidence suggesting that MAP kinase-mediated Sir3p hyperphosphorylation, which plays a role in regulating silencing in response to certain stress conditions, is not involved in high temperature-induced strengthening of silencing. In addition, Pnc1p, a positive regulator of Sir2p activity, plays no role in thermal regulation of silencing. Therefore, growth temperature regulates transcriptional silencing by a novel mechanism. PMID- 15544801 TI - Highly ordered supra-molecular organization of the mycobacterial lipoarabinomannans in solution. Evidence of a relationship between supra molecular organization and biological activity. AB - The complex mycobacterial mannosylated lipoarabinomannans (ManLAMs) are currently considered to be the major virulence factors of the pathogenic Mycobacterium tuberculosis. The recognition and the interaction of ManLAMs with immune system receptors have been shown to promote M.tuberculosis phagocytosis but also to down regulate the bactericidal immune response of the host in favor of the survival of the pathogenic bacilli. To date these original biological activities were mainly associated to the presence of mannose residues capping the non-reducing ends of the ramified polysaccharide moiety of these complex lipoglycans. However, we demonstrated recently that the molecular recognition of ManLAM terminal mannose units by human pulmonary surfactant protein A (hSP-A) carbohydrate recognition domains depends on the presence of the lipid moiety of the ManLAMs as proposed by Sidobre et al. in 2002. Thus, we investigated the putative role of the ManLAM aglycon moiety. The data presented here, indicate that the hydrophobic aglycon part of ManLAM is associated to a characteristic concentration-dependent supra molecular organization of these complex molecules. Furthermore, we observed that the deacylated ManLAMs or the lipid-free mannosylated arabinomannans, which do not exhibit characteristic ManLAM activities, do not display this supra-molecular organization. These observations strongly suggest that the ManLAMs immunomodulatory activities might be associated to their particular organization. Finally, the determination of the critical micellar concentration of ManLAMs obviously supports the notion that this supra-molecular organization may be responsible for the specific biological activities of these complex molecules. PMID- 15544802 TI - Ligand-induced conformational changes in the Bacillus subtilis chemoreceptor McpB determined by disulfide crosslinking in vivo. AB - Previously, we characterized the organization of the transmembrane (TM) domain of the Bacillus subtilis chemoreceptor McpB using disulfide crosslinking. Cysteine residues were engineered into serial positions along the two helices through the membrane, TM1 and TM2, as well as double mutants in TM1 and TM2, and the extent of crosslinking determined to characterize the organization of the TM domain. In this study, the organization of the TM domain was studied in the presence and absence of ligand to address what ligand-induced structural changes occur. We found that asparagine caused changes in crosslinking rate on all residues along the TM1-TM1' helical interface, whereas the crosslinking rate for almost all residues along the TM2-TM2' interface did not change. These results indicated that helix TM1 rotated counterclockwise and that TM2 did not move in respect to TM2' in the dimer on binding asparagine. Interestingly, intramolecular crosslinking of paired substitutions in 34/280 and 38/273 were unaffected by asparagine, demonstrating that attractant binding to McpB did not induce a "piston-like" vertical displacement of TM2 as seen for Trg and Tar in Escherichia coli. However, these paired substitutions produced oligomeric forms of receptor in response to ligand. This must be due to a shift of the interface between different receptor dimers, within previously suggested trimers of dimers, or even higher order complexes. Furthermore, the extent of disulfide bond formation in the presence of asparagine was unaffected by the presence of the methyl modification enzymes, CheB and CheR, or the coupling proteins, CheW and CheV, demonstrating that these proteins must have local structural effects on the cytoplasmic domain that is not translated to the entire receptor. Finally, disulfide bond formation was also unaffected by binding proline to McpC. We conclude that ligand-binding induced a conformational change in the TM domain of McpB dimers as an excitation signal that is likely propagated within the cytoplasmic region of receptors and that subsequent adaptational events do not affect this new TM domain conformation. PMID- 15544803 TI - Double-stranded DNA stimulates the fibrillation of alpha-synuclein in vitro and is associated with the mature fibrils: an electron microscopy study. AB - Filamentous aggregates formed by alpha-synuclein are a prominent and presumably key etiological factor in Parkinson's and other neurodegenerative diseases characterized by motor disorders. Numerous studies have demonstrated that various environmental and intracellular factors affect the fibrillation properties of alpha-synuclein, e.g. by accelerating the process of assembly. Histones, the major component and constituent of chromatin, interact specifically with alpha synuclein and enhance its fibrillation significantly. Here, we report that another component of chromatin, double-stranded DNA (dsDNA), either linear or supercoiled, also interacts with wild-type alpha-synuclein, leading to a significant stimulation of alpha-synuclein assembly into mature fibrils characterized by a reduced lag phase. In general, the morphology of the fibrils remains unchanged in the presence of linear dsDNA. Electron microscopy reveals that DNA forms various types of complexes upon association with the fibrils at their surface without distortion of the double-helical structure. The existence of these complexes was confirmed by the electrophoresis, which also demonstrated that a fraction of the associated DNA was resistant to digestion by restriction endonucleases. Fibrils assembled from the alpha-synuclein mutants A30P and A53T and the C-terminally truncated variants (encoding amino acid residues 1-108 or 1 124) also form complexes with linear dsDNA. Possible mechanisms and implications of dsDNA-alpha-synuclein interactions are discussed. PMID- 15544804 TI - The human protein translin specifically binds single-stranded microsatellite repeats, d(GT)n, and G-strand telomeric repeats, d(TTAGGG)n: a study of the binding parameters. AB - We have previously identified in human fibroblasts a multisubunit protein (designated PGB) that specifically bound single-stranded G-rich microsatellite DNA sequences. PGB was later found to be identical, or closely related to translin, an octameric protein that bound single-stranded DNA consisting of sequences flanking chromosomal translocations. Here, we report that recombinant translin binds single-stranded microsatellite repeats, d(GT)n, and G-strand telomeric repeats, d(TTAGGG)n, with higher affinities (Kdis approximately = 2 nM and Kdis approximately = 12.5 nM, respectively, in 100 mM NaCl and 25 degrees C) than the affinity with which it binds a prototypical sequence flanking translocation sites (Kdis approximately = 23 nM). Translin also binds d(GT)n and d(TTAGGG)n overhangs linked to double-stranded DNA with equilibrium constants in the nanomolar range. Formation of DNA quadruplexes by the d(TTAGGG)n repeats inhibits their binding to translin. A further study of the binding parameters revealed that the minimal length of d(GT)n and d(TTAGGG)n oligonucleotides that a translin octamer can bind is 11 nucleotides, but that such oligonucleotides containing up to 30 nucleotides can bind only a single translin octamer. However, the oligonucleotides d(GT)27 and d(TTAGGG)9 bind two octamers with negative cooperativity. Translin does not detectably bind single-stranded d(GT)n sequences embedded within double-stranded DNA. Based on our data, we propose that translin might be involved in the control of recombination at d(GT)n.d(AC)n microsatellites and in telomere maintenance. PMID- 15544805 TI - Crystal structure of RecA from Deinococcus radiodurans: insights into the structural basis of extreme radioresistance. AB - The resistance of Deinococcus radiodurans (Dr) to extreme doses of ionizing radiation depends on its highly efficient capacity to repair dsDNA breaks. Dr RecA, the key protein in the repair of dsDNA breaks by homologous recombination, promotes DNA strand-exchange by an unprecedented inverse pathway, in which the presynaptic filament is formed on dsDNA instead of ssDNA. In order to gain insight into the remarkable repair capacity of Dr and the novel mechanistic features of its RecA protein, we have determined its X-ray crystal structure in complex with ATPgammaS at 2.5A resolution. Like RecA from Escherichia coli, Dr RecA crystallizes as a helical filament that is closely related to its biologically relevant form, but with a more compressed pitch of 67 A. Although the overall fold of Dr RecA is similar to E.coli RecA, there is a large reorientation of the C-terminal domain, which in E.coli RecA has a site for binding dsDNA. Compared to E.coli RecA, the inner surface along the central axis of the Dr RecA filament has an increased positive electrostatic potential. Unique amino acid residues in Dr RecA cluster around a flexible beta-hairpin that has also been implicated in DNA binding. PMID- 15544806 TI - Efficient attenuation of stochasticity in gene expression through post transcriptional control. AB - Thermal fluctuations can lead to significant, unpredictable concentration changes in intracellular molecules, potentially disrupting the functioning of cellular networks and challenging cellular efficiency. Biochemical systems might therefore be expected to have evolved network architectures and motifs that limit the effects of stochastic disturbances. During gene expression itself, stochasticity, or "noise", in protein concentrations is believed to be determined mostly by mRNA, rather than protein, levels. Here, we demonstrate in silico, and analytically, how a number of commonly occurring network architectures in bacteria use mRNA to efficiently attenuate fluctuations. Genes coded in operons share mRNA, which we show generates strongly correlated expression despite multiple ribosome binding sites. For autogeneous control, we provide general analytic expressions using Langevin theory, and demonstrate that negative translational feedback has a much greater efficiency at reducing stochasticity than negative transcriptional feedback. Using the ribosomal proteins as an example, we also show that translational, rather than transcriptional, feedback best coordinates gene expression during assembly of macromolecular complexes. Our findings suggest that selection of a gene controlled post-transcriptionally may be for the resulting low stochasticity in its expression. Such low noise genes can be speculated to play a central role in the local gene network. PMID- 15544807 TI - Protein biophysical properties that correlate with crystallization success in Thermotoga maritima: maximum clustering strategy for structural genomics. AB - Cost and time reduction are two of the driving forces in the development of new strategies for protein crystallization and subsequent structure determination. Here, we report the analysis of the Thermotoga maritima proteome, in which we compare the proteins that were successfully expressed, purified and crystallized versus the rest of the proteome. This set of almost 500 proteins represents one of the largest, internally consistent, protein expression and crystallization datasets available. The analysis shows that individual parameters, such as isoelectric point, sequence length, average hydropathy, low complexity regions (SEG), and combinations of these biophysical properties for crystallized proteins define a distinct subset of the T. maritima proteome. The distribution profiles of the various biophysical properties in the expression/crystallization set are then used to extract rules to improve target selection and improve the efficiency and output of structural genomics, as well as general structural biology efforts. PMID- 15544808 TI - The D2 period of collagen II contains a specific binding site for the human discoidin domain receptor, DDR2. AB - The human discoidin domain receptors (DDRs), DDR1 and DDR2, are expressed widely and, uniquely among receptor tyrosine kinases, activated by the extracellular matrix protein collagen. This activation is due to a direct interaction of collagen with the DDR discoidin domain. Here, we localised a specific DDR2 binding site on the triple-helical region of collagen II. Collagen II was found to be a much better ligand for DDR2 than for DDR1. As expected, DDR2 binding to collagen II was dependent on triple-helical collagen and was mediated by the DDR2 discoidin domain. Collagen II served as a potent stimulator of DDR2 autophosphorylation, the first step in transmembrane signalling. To map the DDR2 binding site(s) on collagen II, we used recombinant collagen II variants with specific deletions of one of the four repeating D periods. We found that the D2 period of collagen II was essential for DDR2 binding and receptor autophosphorylation, whereas the D3 and D4 periods were dispensable. The DDR2 binding site on collagen II was further defined by recombinant collagen II-like proteins consisting predominantly of tandem repeats of the D2 or D4 period. The D2 construct, but not the D4 construct, mediated DDR2 binding and receptor autophosphorylation, demonstrating that the D2 period of collagen II harbours a specific DDR2 recognition site. The discovery of a site-specific interaction of DDR2 with collagen II gives novel insight into the nature of the interaction of collagen II with matrix receptors. PMID- 15544809 TI - Crystal structures of five bovine chymotrypsin complexes with P1 BPTI variants. AB - The bovine chymotrypsin-bovine pancreatic trypsin inhibitor (BPTI) interaction belongs to extensively studied models of protein-protein recognition. The accommodation of the inhibitor P1 residue in the S1 binding site of the enzyme forms the hot spot of this interaction. Mutations introduced at the P1 position of BPTI result in a more than five orders of magnitude difference of the association constant values with the protease. To elucidate the structural aspects of the discrimination between different P1 residues, crystal structures of five bovine chymotrypsin-P1 BPTI variant complexes have been determined at pH 7.8 to a resolution below 2 A. The set includes polar (Thr), ionizable (Glu, His), medium-sized aliphatic (Met) and large aromatic (Trp) P1 residues and complements our earlier studies of the interaction of different P1 side-chains with the S1 pocket of chymotrypsin. The structures have been compared to the complexes of proteases with similar and dissimilar P1 preferences, including Streptomyces griseus proteases B and E, human neutrophil elastase, crab collagenase, bovine trypsin and human thrombin. The S1 sites of these enzymes share a common general shape of significant rigidity. Large and branched P1 residues adapt in their complexes similar conformations regardless of the polarity and size differences between their S1 pockets. Conversely, long and flexible residues such as P1 Met are present in the disordered form and display a conformational diversity despite similar inhibitory properties with respect to most enzymes studied. Thus, the S1 specificity profiles of the serine proteases appear to result from the precise complementarity of the P1-S1 interface and minor conformational adjustments occurring upon the inhibitor binding. PMID- 15544810 TI - Characterization of the structure and the amyloidogenic properties of the Josephin domain of the polyglutamine-containing protein ataxin-3. AB - Expansion of the polyglutamine (polyQ) region in the protein ataxin-3 is associated with spinocerebellar ataxia type 3, an inherited neurodegenerative disorder that belongs to the family of polyQ diseases. Increasing evidence indicates that protein aggregation and fibre formation play an important role in these pathologies. In a previous study, we determined the domain architecture of ataxin-3, suggesting that it comprises a globular domain, named Josephin, and a more flexible C-terminal region, that includes the polyQ tract. Here, we have characterised for the first time the biophysical properties of the isolated Josephin motif, showing that it is an autonomously folded unit and that it has no significant interactions with the C-terminal region. Study of its thermodynamic stability indicates that Josephin has an intrinsic tendency to aggregate and forms temperature-induced fibrils similar to those described for expanded ataxin 3. We show that, under destabilising conditions, the behaviours of the isolated Josephin domain and ataxin-3 are extremely similar. Our data therefore strongly suggest that the stability and aggregation properties of non-expanded ataxin-3 are determined by those of the Josephin domain, which is sufficient to reproduce the behaviour of the full-length protein. Our data support a mechanism in which the thermodynamic stability of ataxin-3 is governed by the properties of the Josephin domain, but the presence of an expanded polyQ tract increases dramatically the protein's tendency to aggregate. PMID- 15544811 TI - Alzheimer's disease Abeta peptide fragment 10-30 forms a spectrum of metastable oligomers with marked preference for N to N and C to C monomer termini proximity. AB - Oligomers of Abeta peptide have been indicated recently as a possible main causative agent of Alzheimer's disease. However, information concerning their structural properties is very limited. Here Abeta oligomers are studied by non covalent complexes mass spectrometry and disulfide rearrangement. As a model molecule, an Abeta fragment spanning residues 10-30 (Abeta10-30) has been used. This model peptide is known to contain the core region responsible for Abeta aggregation to fibrils. Non-covalent complexes mass spectrometry indicates that, at neutral pH, monomers are accompanied by oligomers up to hexamers of gradually decreasing population. H-2H exchange studies and direct monomer exchange rate measurements with the use of 15N labeled peptides and mass spectrometry show a fast exchange of monomeric units between oligomers. Disulfide exchange studies of cysteine tagged Abeta10-30 and its mutant show proximity of N-N and C-C termini of monomers in oligomers. The presented data underscore a dynamic character for pre-nucleation forms of Abeta, however, with a marked tendency for parallel strand orientation in oligomers. PMID- 15544812 TI - Foldon, the natural trimerization domain of T4 fibritin, dissociates into a monomeric A-state form containing a stable beta-hairpin: atomic details of trimer dissociation and local beta-hairpin stability from residual dipolar couplings. AB - The C-terminal domain of T4 fibritin (foldon) is obligatory for the formation of the fibritin trimer structure and can be used as an artificial trimerization domain. Its native structure consists of a trimeric beta-hairpin propeller. At low pH, the foldon trimer disintegrates into a monomeric (A-state) form that has similar properties as that of an early intermediate of the trimer folding pathway. The formation of this A-state monomer from the trimer, its structure, thermodynamic stability, equilibrium association and folding dynamics have been characterized to atomic detail by modern high-resolution NMR techniques. The foldon A-state monomer forms a beta-hairpin with intact and stable H-bonds that is similar to the monomer in the foldon trimer, but lacks a defined structure in its N and C-terminal parts. Its thermodynamic stability in pure water is comparable to designed hairpins stabilized in alcohol/water mixtures. Details of the thermal unfolding of the foldon A-state have been characterized by chemical shifts and residual dipolar couplings (RDCs) detected in inert, mechanically stretched polyacrylamide gels. At the onset of the thermal transition, uniform relative changes in RDC values indicate a uniform decrease of local N-HN and Calpha-Halpha order parameters for the hairpin strand residues. In contrast, near turn residues show particular thermal stability in RDC values and hence in local order parameters. This coincides with increased transition temperatures of the beta-turn residues observed by chemical shifts. At high temperatures, the RDCs converge to non-zero average values consistent with predictions from random chain polymer models. Residue-specific deviations above the unfolding transition reveal the persistence of residual order around proline residues, large hydrophobic residues and at the beta-turn. PMID- 15544813 TI - Solution structure of the dimeric SAM domain of MAPKKK Ste11 and its interactions with the adaptor protein Ste50 from the budding yeast: implications for Ste11 activation and signal transmission through the Ste50-Ste11 complex. AB - Ste11, a homologue of mammalian MAPKKKs, together with its binding partner Ste50 works in a number of MAPK signaling pathways of Saccharomyces cerevisiae. Ste11/Ste50 binding is mediated by their sterile alpha motifs or SAM domains, of which homologues are also found in many other intracellular signaling and regulatory proteins. Here, we present the solution structure of the SAM domain or residues D37-R104 of Ste11 and its interactions with the cognate SAM domain containing region of Ste50, residues M27-Q131. NMR pulse-field-gradient (PFG) and rotational correlation time measurements (tauc) establish that the Ste11 SAM domain exists predominantly as a symmetric dimer in solution. The solution structure of the dimeric Ste11 SAM domain consists of five well-defined helices per monomer packed into a compact globular structure. The dimeric structure of the SAM domain is maintained by a novel dimer interface involving interactions between a number of hydrophobic residues situated on helix 4 and at the beginning of the C-terminal long helix (helix 5). The dimer structure may also be stabilized by potential salt bridge interactions across the interface. NMR H/2H exchange experiments showed that binding of the Ste50 SAM to the Ste11 SAM very likely involves the positively charged extreme C-terminal region as well as exposed hydrophobic patches of the dimeric Ste11 SAM domain. The dimeric structure of the Ste11 SAM and its interactions with the Ste50 SAM may have important roles in the regulation and activation of the Ste11 kinase and signal transmission and amplifications through the Ste50-Ste11 complex. PMID- 15544814 TI - Non-linear effects of temperature and urea on the thermodynamics and kinetics of folding and unfolding of hisactophilin. AB - Extensive measurements and analysis of thermodynamic stability and kinetics of urea-induced unfolding and folding of hisactophilin are reported for 5-50 degrees C, at pH 6.7. Under these conditions hisactophilin has moderate thermodynamic stability, and equilibrium and kinetic data are well fit by a two-state transition between the native and the denatured states. Equilibrium and kinetic m values decrease with increasing temperature, and decrease with increasing denaturant concentration. The betaF values at different temperatures and urea concentrations are quite constant, however, at about 0.7. This suggests that the transition state for hisactophilin unfolding is native-like and changes little with changing solution conditions, consistent with a narrow free energy profile for the transition state. The activation enthalpy and entropy of unfolding are unusually low for hisactophilin, as is also the case for the corresponding equilibrium parameters. Conventional Arrhenius and Eyring plots for both folding and unfolding are markedly non-linear, but these plots become linear for constant DeltaG/T contours. The Gibbs free energy changes for structural changes in hisactophilin have a non-linear denaturant dependence that is comparable to non linearities observed for many other proteins. These non-linearities can be fit for many proteins using a variation of the Tanford model, incorporating empirical quadratic denaturant dependencies for Gibbs free energies of transfer of amino acid constituents from water to urea, and changes in fractional solvent accessible surface area of protein constituents based on the known protein structures. Noteworthy exceptions that are not well fit include amyloidogenic proteins and large proteins, which may form intermediates. The model is easily implemented and should be widely applicable to analysis of urea-induced structural transitions in proteins. PMID- 15544815 TI - Insertion kinetics of a denatured alpha helical membrane protein into phospholipid bilayer vesicles. AB - Membrane protein folding has suffered from a lack of detailed kinetic studies, particularly with regard to the insertion of denatured protein into lipid bilayers. We present a detailed in vitro kinetic study of the association of a denatured, transmembrane alpha helical protein with lipid vesicles. The mechanism of folding of Escherichia coli diacylglycerol kinase from a partially denatured state in urea has been investigated. The protein associates with lipid vesicles to give a protein, vesicle complex with an apparent association constant of 2 x 10(6) M(-1) s(-1). This association rate approaches the diffusion limit of the protein, vesicle reaction. The association of the protein with lipid vesicles is followed by a slower process occurring at observed rate of 0.031 s(-1), involving insertion into the bilayer and generation of a functional oligomer of diacylglycerol kinase. Protein aggregation competes with vesicle insertion. The urea-denatured protein monomers begin to aggregate as soon as the urea is diluted. This aggregation is faster than the association of the protein with vesicles so that most protein aggregates before it inserts into a vesicle. Increasing the vesicle concentration favours insertion of protein monomers, but at high vesicle concentrations monomers are primarily in separate vesicles and do not associate to form functional oligomers. Irreversible aggregation limits the yield of functional protein, while the data also suggest that lipid vesicles can reverse another aggregation reaction, leading to the recovery of correctly folded protein. PMID- 15544816 TI - Structural stability of oligomeric chaperonin 10: the role of two beta-strands at the N and C termini in structural stabilization. AB - Chaperonin 10 (cpn10) is a well-conserved subgroup of the molecular chaperone family. GroES, the cpn10 from Escherichia coli, is composed of seven 10kDa subunits, which form a dome-like oligomeric ring structure. From our previous studies, it was found that GroES unfolded completely through a three-state unfolding mechanism involving a partly folded monomer and that this reaction was reversible. In order to study whether these unfolding-refolding characteristics were conserved in other cpn10 proteins, we have examined the structural stabilities of cpn10s from rat mitochondria (RatES) and from hyperthermophilic eubacteria Thermotoga maritima (TmaES), and compared the values to those of GroES. From size-exclusion chromatography experiments in the presence of various concentrations of Gdn-HCl at 25 degrees C, both cpn10s showed unfolding-refolding characteristics similar to those of GroES, i.e. two-stage unfolding reactions that include formation of a partially folded monomer. Although the partially folded monomer of TmaES was considerably more stable compared to GroES and RatES, it was found that the overall stabilities of all three cpn10s were achieved significantly by inter-subunit interactions. We studied this contribution of inter-subunit interactions to overall stability in the GroES heptamer by introducing a mutation that perturbed subunit association, specifically the interaction between the two anti-parallel beta-strands at the N and C termini of this protein. From analyses of the mutants' stabilities, it was revealed that the anti-parallel beta-strands at the subunit interface are crucial for subunit association and stabilization of the heptameric GroES protein. PMID- 15544817 TI - Network analysis of protein structures identifies functional residues. AB - Identifying active site residues strictly from protein three-dimensional structure is a difficult task, especially for proteins that have few or no homologues. We transformed protein structures into residue interaction graphs (RIGs), where amino acid residues are graph nodes and their interactions with each other are the graph edges. We found that active site, ligand-binding and evolutionary conserved residues, typically have high closeness values. Residues with high closeness values interact directly or by a few intermediates with all other residues of the protein. Combining closeness and surface accessibility identified active site residues in 70% of 178 representative structures. Detailed structural analysis of specific enzymes also located other types of functional residues. These include the substrate binding sites of acetylcholinesterases and subtilisin, and the regions whose structural changes activate MAP kinase and glycogen phosphorylase. Our approach uses single protein structures, and does not rely on sequence conservation, comparison to other similar structures or any prior knowledge. Residue closeness is distinct from various sequence and structure measures and can thus complement them in identifying key protein residues. Closeness integrates the effect of the entire protein on single residues. Such natural structural design may be evolutionary maintained to preserve interaction redundancy and contribute to optimal setting of functional sites. PMID- 15544818 TI - Neurospora crassa CyPBP37: a cytosolic stress protein that is able to replace yeast Thi4p function in the synthesis of vitamin B1. AB - Recently, we identified CyPBP37 of Neurospora crassa as a binding partner of cyclophilin41. CyPBP37 function had not yet been described, although orthologs in other organisms have been implicated in the biosynthesis of the thiazole moiety of thiamine (vitamin B1) and/or stress-related pathways. Here, CyPBP37 is characterized as an abundant cytosolic protein with a functional NAD-binding site. Saccharomyces cerevisiae mutants lacking Thi4p (the CyPBP37 ortholog) are auxotrophic for vitamin B1 (thiamine) but can grow in the presence of the thiazole moiety of thiamine, suggesting a role for Thi4p in the biosynthesis of thiazole. N.crassa CyPBP37 is able to functionally replace Thi4p in yeast thiazole synthesis. Cellular fractionation studies revealed that Thi4p is a cytosolic protein in S.cerevisiae, like its ortholog CyPBP37 in N.crassa. This implies that thiamine synthesis takes place in the cytosol of both organisms and not in the mitochondria, as suggested. The expression of CyPBP37 and Thi4p is repressed by thiamine but not by thiazole in the growth medium. In addition to its function in thiazole synthesis, CyPBP37 is a stress-inducible protein. N.crassa cyclophilin41 can chaperone the folding of CyPBP37, its own binding partner. PMID- 15544820 TI - Physical examination of the shoulder in the primary care setting. AB - Shoulder problems are extremely common reasons for visiting a primary care physician. To make a correct diagnosis, an appropriate physical examination must be performed. This article describes a systematic approach to the physical examination, correlating the various maneuvers and tests with the anatomy and pathology of the shoulder. The steps in this examination are: inspection, observation, palpation, manual muscle testing, and special testing. With this type of systematic approach, the physician is more likely to arrive at an accurate diagnosis. PMID- 15544821 TI - Shoulder impingement syndrome. AB - This article deals with the diagnosis and treatment of shoulder impingement syndrome. PMID- 15544822 TI - Rotator cuff injuries and treatment. AB - Rotator cuff problems are among the most commonly encountered disorders of the shoulder, and are commonly seen by primary care physicians. Their exact mechanism for susceptibility to injury remains unclear; however, with an understanding of the shoulder's anatomy and biomechanics, we are better able to treat the insults incurred on the cuff. Early recognition, proper treatment, and appropriate follow up may expedite healing and prevent the occurrence of further injury or complications. PMID- 15544823 TI - Superior labrum, anterior-posterior lesions and biceps injuries: diagnostic and treatment considerations. AB - Proximal biceps tendon rupture, biceps tendinitis, and medical biceps subluxation are injuries that occur alone and in association with other pathologic conditions of the shoulder. The literature describes various strategies for imaging and treating these biceps tendon complex injuries but offers no clear consensus on a gold-standard approach. The keys to successful treatment are an accurate diagnosis, identification of associated pathologic conditions, and assessment of functional limitations of the injury. PMID- 15544824 TI - The acromioclavicular joint. AB - Acute injuries of the AC joint have a limited differential diagnosis. A thorough examination and appropriate radiographs can identify nearly all problems while ruling out tumors as a causal factor. AC injury frequently occurs with other fractures, dislocations, or soft tissue injury around the shoulder. The single unifying diagnosis one searches for in medical conditions is frequently absent in musculoskeletal injury. For effective management of AC separations, the primary care physician need only distinguish type I, II, and III injuries from surgically treated type IV, V, and VI injuries. Types I, II, and III injuries should be treated with pain control and progressive rehabilitation as described previously. Patients who have types IV, V, and VI injuries should be referred to an orthopedic surgeon. Chronic injuries of the AC joint are also clearly diagnosed by appropriate physical examination and radiographs. Steroid injections can aid in the diagnosis and management of these conditions. Injury to one shoulder component predisposes to other shoulder injuries. A thorough examination will reveal these other associated abnormalities that may not be part of the presenting complaint. With practice in the examination of the shoulder and intra articular injections, the primary care physician can effectively treat most common AC conditions. Only after conservative measures fail is it necessary to refer the patient for surgical management. PMID- 15544825 TI - Shoulder instability. AB - Shoulder instability can result from acute injury or repetitive overhead activity. Once the injury is identified, the initial course of treatment is aggressive rehabilitation. Structural injuries can occur but may not be clinically significant. Should a thorough scapular stabilization and rotator cuff strengthening program fail, consultation with an orthopedic surgeon to consider further imaging or possible intervention is appropriate. As the physician's study of approaches to the first-time dislocator continues, they will be better informed of reasonable options to offer the athlete. PMID- 15544826 TI - Physical examination of the knee. AB - Regardless of whether knee pain is acute or chronic in presentation, the primary care physician must be confident and practiced in his or her musculoskeletal examination skills to perform a thorough assessment. Keeping in mind the types of problems that tend to occur in different age groups and in certain patient populations with particular risk factors, a preconceived differential diagnosis list should be present in the physician's mind. In most cases, the history and physical examination findings are sufficient to formulate a definitive diagnosis. Plain radiography (multiple views) and laboratory studies if indicated may narrow the differential. Advanced imaging studies will affirm the clinical findings. Referral may be necessary for surgical management, particularly in pediatric and unstable orthopedic cases (eg, suspected growth plate injury, compartment syndromes, displaced or intra-articular fractures). Consultation should always be readily available if questions arise on preliminary clinical work-up and management. PMID- 15544827 TI - Patellar tendonitis and patellar dislocations. AB - Common sports injuries, such as patellar tendonitis and patellar dislocation, can be treated either surgically or with rehabilitation and physical therapy. Most patients with patellar tendonitis will respond well to conservative measures; however, some recalcitrant cases will require surgical intervention. To date, the literature is not able objectively to identify the patients best suited to surgery. Likewise, in the case of patellar dislocations, it is still unclear which patients respond best to conservative therapy and which respond best to surgical treatment, although evidence continues to accumulate. A practical approach to this problem can be deduced from the available evidence, but more well-designed clinical trials are needed for the establishment of definitive treatment protocols. PMID- 15544828 TI - Anterior and posterior cruciate ligament injuries. AB - Knee pain is a common complaint in the primary care setting, and ACL and PCL injuries are common causes of knee pain. Therefore, it is important for the primary care physician to be skilled in the diagnosis and initial management of these injuries and to be aware of potential associated knee injuries. By understanding the history, mechanism of injury, physical examination maneuvers, and imaging modalities related to ACL and PCL injuries, primary care physicians can prepare themselves to make an accurate diagnosis. Then they can counsel the patients on the available treatments, initiate conservative treatment if appropriate, and facilitate an effective referral to a primary care sports medicine physician or orthopedic surgeon. PMID- 15544829 TI - Medial and lateral collateral injuries: prognosis and treatment. AB - This article presents a comprehensive review of medial and lateral knee ligament injuries. It reviews the basic anatomy, physical examination, imaging techniques, and current treatment of medial and lateral knee injuries. It includes a step-by step guide to proper physical examination of the knee as well as MRI images. Current treatment of medial and lateral knee injuries is extensively reviewed to aid the primary care physician in diagnosis and provision of care. PMID- 15544830 TI - Patellofemoral pain syndrome: evaluation and treatment. AB - Patellofemoral pain syndrome is common among athletes and non-athletes. It results from an imbalance of forces acting on the patellofemoral joint, which leads to increased strain on the peripatellar soft tissues, increased patellofemoral joint stress, or both. The most important risk factors are overuse, quadriceps weakness, and soft-tissue tightness. In most cases, the etiology is multifactorial. A careful history and targeted physical examination will confirm the diagnosis and determine the most appropriate treatment. A physical therapy program that employs quadriceps strengthening, manual stretching of the lateral patellar soft-tissue structures, patellar taping, and biofeedback is successful in the majority of cases. Surgery maybe required for the few patients who do not respond to nonoperative management. PMID- 15544831 TI - Soft-tissue injuries of the lower extremity. AB - A diverse set of conditions affect the lower extremities in athletes. Good history taking and knowledge of the anatomy of the affected areas can aid in accurate diagnosis. Most conditions can be treated conservatively with great success. Familiarity with rehabilitation strategies can assist physicians in working with physical therapists. New advances, especially in the therapy of plantar fasciitis, may aid physicians in more rapid treatment of conditions of overuse. Surgery can provide definitive relief from symptoms for athletes with whom conservative measures have failed. PMID- 15544832 TI - The ankle examination. AB - Ankle injuries are among the most common musculoskeletal complaints a physician will see. A thorough understanding of the bony, ligamentous, and musculotendinous anatomy and an appreciation of the biomechanics of the ankle and mechanisms of injury are important to the accurate assessment of ankle injuries. PMID- 15544833 TI - Achilles tendonopathy and tendon rupture: conservative versus surgical management. AB - Injuries to the Achilles tendon are common in primary care. Insertional tendonitis, retrocalcaneal bursitis, and paratenonitis are acute injuries usually treated conservatively with rest, ice, anti-inflammatory measures, and physical rehabilitation. Causative factors such as improper training or biomechanical abnormalities must be corrected to prevent reoccurrence. Achilles tendinosis is a chronic condition that does not always cause clinical symptoms. When symptoms occur, they are thought to be due to microtrauma or progressive failure resulting in inflammation. Again, conservative treatment usually relieves symptoms, but treatment may be prolonged. Surgical treatment may occasionally be recommended. With rupture of the Achilles, there exists some controversy regarding the advantage of conservative versus surgical management. Treatment should be based on individual patient considerations and expectations. PMID- 15544834 TI - Ankle problems masquerading as sprains. AB - The most common ankle injury is a ligamentous sprain, usually involving the lateral side. This is usually treated very successfully with conservative management. Those patients who have continued complaints despite such management should be suspected of having something more than a simple sprain. The diagnostic entities discussed herein should always be part of the differential diagnosis of a sprained ankle, so that when one of these injuries is initially missed, the examining physician will be able to re-examine and eventually diagnose and treat these injuries properly. They should be managed appropriately as soon as possible for the best outcome. Needless delays should be avoided. PMID- 15544835 TI - Effect of reserpine on the generation of the chromogranin A-derived neuropeptide WE-14 in rat oxyntic mucosa. AB - WE-14, a post-translational product of the neuroendocrine protein chromogranin A (CgA), is generated in distinct subpopulations of endocrine cells. The objective of this study was to investigate the generation of WE-14 in the endocrine cell types of the oxyntic mucosa of the stomach, after treatment with reserpine, an irreversible inhibitor of vesicular monoamine uptake 2 (VMAT2). Reserpine (10 mg/kg) was administered subcutaneously and tissue analysed 1, 3, 5 and 18 h following treatment. The oxyntic mucosa was analysed immunohistochemically employing a site-specific WE-14 antiserum, a region-specific CgA antiserum and an antiserum against histidine decarboxylase (HDC), a marker of the histamine producing ECL cells in the oxyntic mucosa. The number of oxyntic endocrine cells exhibiting WE-14 immunostaining increased more than 100-fold 18 h after reserpine administration relative to vehicle treated controls. Double immunostaining with HDC revealed that most, but not all, of the WE-14 positive cells were ECL cells. These results suggest that reserpine has the ability to influence the post translational processing of CgA to generate WE-14 in rat stomach ECL cells, presumably as a consequence of reduced VMAT2-driven accumulation of histamine. PMID- 15544836 TI - Estrogen upregulates renal angiotensin II AT1 and AT2 receptors in the rat. AB - We studied renal AT1 and AT2 receptors in male, female, ovariectomized and ovariectomized-estrogen-treated Wistar-Hanover and Wistar-Kyoto rats. AT1 receptors and AT1A receptor mRNA predominated, with no significant differences between males and females. AT2 receptor expression was restricted in female rats to the capsule, the transition zone between outer and inner medulla, the endothelium lining the papilla, and arcuate arteries and veins. There were no AT2 receptors in male rats, while male mice express substantial numbers of estrogen dependent AT2 receptors. Arcuate arteries and veins expressed AT1B mRNA in males and females, and AT2 mRNA in females only. AT1 receptor and AT2 receptor expression were estrogen-dependent, with increases in AT1 and AT2 receptor expression after estrogen treatment in ovariectomized rats. Estrogen treatment increased prostaglandin E2 (PGE2) and cGMP concentrations in the renal medulla, and eNOS expression in cortical arteries. In rodents, expression of renal Angiotensin II receptor types is estrogen-dependent, with significant species, strain and area differences. Our results support an important role for AT2 receptors in the regulation of renal function and in the protective effects of estrogen in the kidney. PMID- 15544837 TI - Monocytes of allergics and non-allergics produce, store and release the neurotrophins NGF, BDNF and NT-3. AB - INTRODUCTION: Recent studies have shown that neurotrophins (NTs) are involved in inflammatory processes. Elevated plasma levels of NTs were found allergic diseases with the highest levels in allergic asthma. However, the exact cellular sources involved in the regulation and release of neurotrophins in allergic inflammation are still not well defined. OBJECTIVE: The aim of this study was to assess whether monocytes of allergic and non-allergic subjects produce, store and release the neurotrophins NGF, BDNF and NT-3. METHODS: Monocytes of allergic and non-allergic donors were purified by immunomagnetic selection. APAAP-staining for the presence of NTs and their receptors was performed. RT-PCR and Western blot evaluated the production and storage of NTs. Monocytes were incubated and supernatants were collected for measurement of neurotrophic factors after stimulation with lipopolysaccharide (LPS) as inflammatory stimulus. The neurotrophin content in lysates and cell culture supernatants was determined by ELISA. RESULTS: Human monocytes express the neurotrophins NGF, BDNF and NT-3 but also their specific receptors TrkA, TrkB and TrkC. RT-PCR amplification of isolated mRNA demonstrated expression of the examined neurotrophins. Proteins were detectable by Western blot. NTs were found in the monocyte lysates and supernatants at different levels in allergic and non-allergic donors. Cell stimulation with LPS leads to release of NGF and NT3. CONCLUSIONS: Monocytes, produce, store and release NGF, BDNF and NT-3. They are a possible source of elevated neurotrophin levels found in allergy and asthma. PMID- 15544839 TI - Interactions of galanin and opioids in nociceptive modulation in the arcuate nucleus of hypothalamus in rats. AB - The fact that galanin, beta-endorphin and their receptors are present in the arcuate nucleus of hypothalamus (ARC), coupled with our previous observation that both beta-endorphin and galanin play antinociceptive roles in pain modulation in the ARC, made it of interest to study their interactions. The hindpaw withdrawal latency (HWL) in response to noxious thermal and mechanical stimulation was assessed by the hot-plate test and the Randall Selitto Test. We showed that the antinociceptive effect induced by intra-ARC injection of galanin was dose dependently attenuated by the following intra-ARC injection of naloxone. Furthermore, intra-ARC administration of the selective mu-opioid receptor antagonist beta-funaltrexamine (beta-FNA) attenuated the increased HWL induced by intra-ARC injection of galanin in a dose-dependent manner, while the delta-opioid receptor antagonist naltrindole or the kappa-opioid receptor antagonist nor binaltorphimine (nor-BNI) did not. Moreover, intra-ARC injection of a galanin receptor antagonist galantide attenuated intraperitoneal morphine-induced increases in HWLs. These results demonstrate that the antinociceptive effect of galanin was related to the opioid system, especially mu-opioid receptor was involved in, and that systemic morphine induced antinociception involves galanin in the ARC. PMID- 15544838 TI - Effect of acetic acid or trypsin application on rat colonic motility in vitro and modulation by two synthetic fragments of chromogranin A. AB - The hypothesis that Chromogranin A (CgA)-derived peptides are involved in mechanisms modulating altered colonic motility was tested. Rat distal colonic strips were studied using an organ bath technique. Acetic acid (AA)-induced effects were characterized on spontaneous mechanical activities (SMA) in the presence of CgA4-16 or CgA47-66. In preparations with mucosa, AA induced a transient hyperactivity followed by a decrease in tone. The first phase is sensitive to tetrodotoxin (TTX) and capsaicin. The second phase was sensitive to BAYK8644 but insensitive to L-nitro-arginine-methyl-ester (L-Name)/apamin together. CgA4-16 or CgA47-66 alone produced no change on SMA. The administration of CgA4-16 prior to AA increased the duration of the excitatory component and reduced tone inhibition. CgA47-66 prior to AA only decreased duration of the excitatory phase. In preparations without mucosa, AA decreased tone. This effect was sensitive to BAYK8644 and CgA4-16. Trypsin decreased basal tone. This effect was suppressed by TTX, BAYK8644 or L-Name/apamin and were reduced by CgA4-16. AA induced effects on rat colonic motility in vitro may be mediated through activation of primary afferents and an action at L-Type calcium channels. CgA derived peptides are shown to decrease AA-induced effects on motility. PMID- 15544840 TI - Overexpression of Bcl-2 inhibits nuclear localization of annexin I during tumor necrosis factor-alpha-mediated apoptosis in porcine renal LLC-PK1 cells. AB - The addition of tumor necrosis factor (TNF)-alpha into the cultured porcine kidney LLC-PK1 cells caused apoptosis concomitantly with caspase-3 activation and the inductions of an endogenous Bcl-2 protein. An SDS-polyacrylamide electrophoretic analysis revealed that a 37-kDa protein in a nuclear fraction was increased during TNF-alpha-induced apoptosis. Partial amino acid sequence of the protein was A-L-T-G-H-L-E-E-V, perfectly matching that of annexin I. Immunocytochemistry revealed that annexin I migrated to the nucleus and/or peri nucleus region upon exposure to TNF-alpha. Overexpression of Bcl-2 proteins inhibited the nuclear localization of annexin I during TNF-alpha-induced apoptosis. Antisense oligodeoxynucleotides complementary to annexin I-inhibited TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labelling) staining in TNF-alpha-treated cells, suggesting that annexin I expression is a possible prerequisite for the induction of apoptosis by the cytokine. Thus, it is first time to show that annexin I is regulated by an anti apoptotic Bcl-2 protein in TNF-alpha-induced renal apoptotic events. PMID- 15544841 TI - Enhanced running wheel activity of both Mch1r- and Pmch-deficient mice. AB - Mch1r-deficient (Mch1r(-/-)) mice are hyperphagic, hyperactive, lean, and resistant to diet-induced obesity. To examine whether the MCH1R is involved in regulating activity-based energy expenditure, we investigated voluntary wheel running (WR) activity of wild-type (WT) and Mch1r(-/-) mice basally, in response to diets with different caloric density and with different feeding schedules. We also evaluated WR activity of mice with ablation of the prepro-MCH gene (Pmch(-/ ) mice). Dark cycle WR activity of Mch1r(-/-) mice fed low fat (LF) chow was increased significantly relative to WT mice. Transition to moderate high-fat (MHF) diet was associated with an increase in nocturnal WR activity in both genotypes. Both Mch1r(-/-) and WT mice exhibited food anticipatory activity (FAA) before the daily scheduled feeding time, indicating that MCH1R is not required for FAA. Naloxone (3 mg/kg, i.p.) suppressed WR activity of both genotypes, suggesting opioid regulation of locomotor activity. WR increased nocturnal dynorphin mRNA levels in Mch1r(-/-) brain. Importantly, Pmch-deficient mice had significantly enhanced WR activity relative to WT controls. These results suggest that endogenous MCH plays an inhibitory role in regulating locomotor activity. In summary, we demonstrated enhanced WR activities in mice lacking either MCH or its cognate receptor. PMID- 15544842 TI - Production and autocrine/paracrine effects of endogenous insulin-like growth factor-1 in rat cardiac fibroblasts. AB - Insulin-like growth factor (IGF)-1 appears to play an important role in cardiac hypertrophy or remodeling. However, the role of endogenous IGF-1 in the growth of cardiac myocytes and fibroblasts remains unclear. This study investigated the major site of the production of cardiac IGF-1 and the local effects of endogenous IGF-1 secreted from cardiac cells. A significant expression of IGF-1 mRNA was found in cultured neonatal and adult rat cardiac fibroblasts, but not in myocytes. In addition, an in vivo examination by in situ hybridization histochemical analyses demonstrated the IGF-1 transcripts in the interstitial fibrotic tissue of the ventricle. Time-dependent secretion of IGF-1 protein was also observed in cultured cardiac fibroblasts. An antibody against IGF-1 decreased collagen synthesis in cardiac fibroblasts under basal conditions. Fibroblast-conditioned medium, as well as exogenous IGF-1, increased protein synthesis in cardiac myocytes, and this increase was inhibited by antibodies against IGF-1 and IGF-1 receptor, IGF binding protein-3, and IGF-1 receptor antagonist. These observations suggest that IGF-1 is produced and released mainly from cardiac fibroblasts and that endogenous IGF-1 promotes collagen synthesis by cardiac fibroblasts and hypertrophy of myocytes as an autocrine and a paracrine factor. Cardiac IGF-1 may function as an endogenous modulator of cardiac hypertrophy or remodeling. PMID- 15544843 TI - Chromogranin A-derived peptides: interaction with the rat posterior cerebral artery. AB - Chromogranin A (CgA), an acidic granule protein of the regulated secretory pathway in the diffuse neuroendocrine system, is postulated to serve as a prohormone for regulatory peptides. Betagranin (rCgA(1-128)), the first N terminal cleavage product of rat CgA, is 87% homologous to the bovine vasostatin I (bCgA(1-76)), previously shown to be vasoinhibitory in bovine resistance arteries. In this study the vasoactivity of homologous rat and bovine peptides was investigated in the rat posterior cerebral artery. Firstly, we examined the interaction of rhodamine (Rh)-labelled bCgA(7-40) and bCgA(47-70) with elements of the arterial wall by fluorescence microscopy. Secondly, rCgA(7-57), bCgA(1 40), bCgA(7-40) and bCgA(47-66) (chromofungin) were studied for effects on arterial tone and intracellular calcium as function of pressure in an arteriograph. Although without dilator or constrictor responses at 60-150 mm Hg, the rat peptide (rCgA(7-57)) evoked a significant delay in the onset of forced dilatation at 170 mm Hg, in contrast to the bovine peptides bCgA(1-40), bCgA(7 40) and bCgA(47-66) (chromofungin). Neither Rh-bCgA(7-40) nor Rh-bCgA(47-70) stained the endothelial layer, while Rh-bCgA(47-70) but not Rh-bCgA(7-40) stained the smooth muscle compartment. Analogously, bCgA(47-66) but not bCgA(7-40) reduced intracellular calcium, however without modifying the myogenic response. Thus, the betagranin peptide rCgA(7-57) and the two bovine chromofungin containing peptides, highly homologous to the corresponding sequence (rCgA(47 66)), affected the rat cerebral artery without vasodilator effects, indicating significant species differences in vasoactivity of the N-terminal domain of CgA. PMID- 15544844 TI - Expression of orexin receptors in the brain and peripheral tissues of the male sheep. AB - Orexins exert their effects through two specific receptors (OX1R and OX2R) that have been found mainly in the brain and also in peripheral tissues of rats and humans. Here, we demonstrate expression of mRNA encoding for ovine OX1R and OX2R in central and peripheral tissues of sheep. Gene expression for orexin receptors in the hypothalamus and the preoptic area was localised by in situ hybridisation. OX1R was detected in arcuate nuclei (ARC), median eminence (ME), the lateral hypothalamic nuclei and preoptic area (POA) and it was scattered along the third ventricle from the paraventricular (PVN) to the ventromedial hypothalamic nuclei (VMH). OX2R was localised in the PVN, ARC, ME, ventral VMH and a small region of the ventral POA. Gene expression for OX1R and OX2R in central and peripheral tissues was analysed using quantitative real time RT-PCR. Both orexin receptor genes were expressed in the hypothalamus, POA, hippocampus, amygdala, olfactory bulb, pineal gland and recess and pituitary gland, whereas only OX1R mRNA was detected in the testis, kidney and adrenal gland. The expression of the genes for orexin receptors in this range of ovine tissues suggests roles for orexins in multiple physiological functions, with actions at both central and peripheral levels. PMID- 15544845 TI - Alteration of renal adrenomedullin and its receptor system in the severely hypertensive rat: effect of diuretic. AB - OBJECTIVE: We investigated the pathophysiological role of the renal adrenomedullin (AM) system, including the ligand, receptor, and amidating activity, in severe hypertensive rats. METHOD: We studied three groups: control Wistar Kyoto rats (WKY), spontaneously hypertensive stroke-prone rats (SHR-SP), and diuretic-treated SHR-SP. We measured AM-mature, active form, and AM-total (active form+inactive form) in plasma and renal tissues, and mRNA levels of AM and AM receptor system components such as calcitonin receptor-like receptor (CRLR), receptor activity-modifying protein (RAMP) 2, and RAMP3 in renal tissues. RESULTS: SHR-SP had higher blood pressure, plasma neurohumoral factors, and lower renal function than WKY. SHR-SP had higher AM-mature and AM-total levels in plasma and renal tissues than WKY. Although the plasma AM-mature/AM-total ratio was similar in the two groups, AM-mature/AM-total ratio in renal tissues was higher in SHR-SP than in WKY. In addition, mRNA levels of AM in the renal cortex and medulla and the mRNA levels of CRLR, RAMP2, and RAMP3 in the renal cortex were higher in SHR-SP than in WKY. Chronic diuretic treatment decreased blood pressure and improved kidney function and neurohumoral factors, with reductions in plasma and renal AM system. CONCLUSION: Upregulation of circulating and renal AM system may modulate pathophysiology in SHR-SP. PMID- 15544846 TI - Substance P mediates AP-1 induction in A549 cells via reactive oxygen species. AB - A common feature in asthma is the induction of reactive oxygen species (ROS) and the AP-1 transcription factor during the inflammatory process. AP-1 induction leads to an increased expression of pro-inflammatory cytokines. Also, higher levels of the pro-inflammatory neuropeptide substance P (SP) have been reported in bronchoalveolar-lavage fluid of asthmatics. Here, the role of SP on ROS induction and the downstream activation of AP-1 in A549 airway epithelial cells was investigated by dichloroflourescein-diacetate method and reporter gene assays. The SP-mediated AP-1 induction was dependent on extracellular calcium and ROS. The likely source of ROS are the mitochondria as rotenone inhibited AP-1 induction and the p47phox subunit of the NADPH oxidase complex, responsible for ROS generation in phagocytotic cells, was not expressed in A549 cells assayed by RT-PCR. This is consistent with results obtained from cells of murine bronchial epithelium, isolated by laser capture microdissection. In summary, this study provides evidence for an SP-mediated induction of AP-1, which may contribute to the expression of pro-inflammatory cytokines. PMID- 15544847 TI - GLP-2 stimulates colonic growth via KGF, released by subepithelial myofibroblasts with GLP-2 receptors. AB - BACKGROUND: Glucagon-like peptide-2 is thought to act as a growth factor for the gut, but the localization of the GLP-2 receptor and mechanism of action on epithelial growth is unclear. METHODS AND RESULTS: We found glucagon-like peptide 2 (GLP-2) receptors mainly on subepithelial myofibroblasts in rat, mouse, marmoset and human small and large intestine by immunohistochemistry and in situ hybridisation. By double labelling we found that these GLP-2 receptor immunoreactive cells also produce smooth muscle actin and keratinocyte growth factor (KGF). By subcutaneous infusion of either GLP-2 alone, GLP-2 plus KGF antibody, KGF antibody alone or saline in mice, we found that KGF antibody abolished the growth promoting effect of GLP-2 in the large intestine, but not in the small intestine. CONCLUSIONS: Our findings suggest that GLP-2 in the gut acts by activating receptors on the subepithelial myofibroblasts, causing the release of growth factors, which in turn stimulate intestinal growth. PMID- 15544848 TI - Endotoxin administration increases hypothalamic somatostatin mRNA through nitric oxide release. AB - Acute inflammation induced by endotoxin (LPS) administration inhibits insulin like growth factor (IGF-I) and growth hormone (GH) secretion. The aim of this study was to elucidate the role of glucocorticoids and nitric oxide (NO) in the effect of LPS on hypothalamic somatostatin gene expression. Adult male Wistar rats were injected with different doses of LPS (5, 10 and 100 microg/kg). Rats received two i.p. injections of LPS (at 17:30 and 8:30 h the following day) and were killed 4 h after the second injection. LPS administration at the dose of 100 microg/kg increased the hypothalamic somatostatin mRNA content, as well as the serum concentrations of corticosterone. Glucocorticoids do not seem to be involved in LPS-induced increase in hypothalamic somatostatin mRNA since adrenalectomy did not prevent this effect. In order to analyze the possible effect of NO, aminoguanidine, an inducible nitric oxide synthase inhibitor, was injected (100 mg/kg s.c.) simultaneously with LPS injection. Aminoguanidine administration did not modify somatostatin mRNA in saline injected rats, but it prevented LPS-induced increase in hypothalamic somatostatin mRNA. These data suggest that the stimulatory effect of endotoxin on hypothalamic somatostatin gene expression is not mediated by glucocorticoids, but instead by the increase in NO release. PMID- 15544849 TI - Evidence for the presence of motilin, ghrelin, and the motilin and ghrelin receptor in neurons of the myenteric plexus. AB - Motilin, a 22-amino acid gastrointestinal peptide, and ghrelin, the natural ligand of the growth hormone secretagogue receptor, form a new group of structurally related peptides. Several lines of evidence suggest that motilin and ghrelin are involved in the control of gastrointestinal motility by the activation of receptors on enteric neurons. The aim of this study was to look for the existence of motilin, ghrelin, and their respective receptors in the myenteric plexus of the guinea pig. We used longitudinal muscle/myenteric plexus (LMMP) preparations and cultures of myenteric neurons of the guinea pig ileum, immunohistochemistry, and reverse transcriptase-polymerase chain reaction (RT PCR). Most of the motilin-immunoreactive (IR; 72.8%) and motilin receptor-IR (68.9%) neurons were also positive for neuronal nitric oxide synthase (nNOS), 72.8% and 68.9%, few for choline acetyl transferase (ChAT), 11.4% and 11.9%, respectively. In contrast, ghrelin was mainly colocalized with ChAT (72.2%), and only 3.6% of ghrelin-positive cells showed nNOS-IR in the LMMP. Neither motilin nor the motilin receptor or ghrelin colocalized with calbindin. RT-PCR studies revealed motilin, ghrelin, and ghrelin receptor mRNA transcripts in LMMP preparations and in cultured myenteric neurons. In conclusion, this study, for the first time, provides direct evidence for the existence of motilin and ghrelin in myenteric neurons and suggests that both peptides may play a role in the activation of the enteric nervous system and hence in the regulation of gastrointestinal motility. PMID- 15544850 TI - Altered renal response to acute volume expansion in transgenic rats harboring the human tissue kallikrein gene. AB - The renal response to acute volume expansion was investigated in transgenic (TGR) rats harboring the human tissue kallikrein gene. After a primer injection of 0.9% NaCl (3 ml/100 g, i.v), Sprague-Dawley (SD) or TGR rats received a continuous infusion of 0.9% NaCl (15 microl/100 g/min, i.a.) through a catheter placed into the carotid artery. Acute volume expansion was produced by a second injection of 0.9% NaCl (2 ml/100 g, i.v.) 65 min after the first injection. Plasma vasopressin (AVP) and atrial natriuretic peptide (ANP) concentration was measured before and within 10 min of volume expansion. TGR animals presented a blunted response to acute volume expansion evidenced by an attenuated increase in total and fractional water and sodium excretion. Before or after volume expansion, plasma AVP and ANP did not differ between SD and TGR. Pre-treatment with the BK-B2 antagonist HOE-140 (7.5 microg/100 g. i.a) partially improved the renal response of TGRs and severely blunted the response in SD rats. These data show that TGR (hKLK1) rats have an impaired renal response to acute volume expansion that can not be accounted for by changes in the release of AVP or ANP. PMID- 15544851 TI - Atrial natriuretic factor stimulates renal dopamine uptake mediated by natriuretic peptide-type A receptor. AB - To determine the effects of atrial natriuretic factor (ANF) on renal dopamine (DA) metabolism, 3H-DA and 3H-L-DOPA uptake by renal tubular cells was measured in experiments carried out in vitro in Sprague-Dawley rats. The receptor type involved was also analyzed. The results indicate that ANF increased at 30 min, DA uptake in a concentration-response fashion having 10 pM ANF as the threshold concentration. Conversely, the uptake of the precursor L-DOPA was not modified by the peptide. ANF effects were observed in tissues from external and juxtamedullar cortex and inner medulla. On this basis, 100 nM ANF was used to continue the studies in external cortex tissues. DA uptake was characterized as extraneuronal uptake, since 100 microM hydrocortisone blocked ANF-induced increase of DA uptake. Renal DA uptake was decreased at 0 degrees C and in sodium-free medium. The effects of ANF in these conditions were not present, confirming that renal DA uptake is mediated by temperature- and sodium-dependent transporters and that the peptide requires the presence of the ion to exhibit its actions on DA uptake. The biological natriuretic peptide type A receptor (NPR-A) mediates ANF effects, since 100 nM anantin, a specific blocker, reversed ANF-dependent increase of DA uptake. The natriuretic peptide type C receptor (NPR-C) is not involved, since the specific analogous 100 nM 4-23 ANF amide has no effect on renal DA uptake and does not alter the effects of 100 nM ANF. In conclusion, ANF stimulates DA uptake by kidney tubular cells. ANF effects are mediated by NPR-A receptors coupled to guanylate cyclase and cGMP as second messenger. The process involved was characterized as a typical extraneuronal uptake, and characterized as temperature and sodium-dependent. This mechanism could be related to DA effects on sodium reabsorption and linked to ANF enhanced natriuresis in the kidney. The increment of endogenous DA into tubular cells, as a consequence of increased DA uptake, would permit D1 receptor recruitment and Na+,K+-ATPase activity inhibition, which results in decreased sodium reabsorption and increased natriuresis. PMID- 15544852 TI - Central administration of a nitric oxide precursor abolishes both the hypothalamic serotonin release and the hypophagia induced by interleukin-1beta in obese Zucker rats. AB - Serotonin-induced anorexia has long been recognized as an important part of the CNS mechanisms controlling energy balance. More recently, interleukin-1beta and nitric oxide have been suggested to influence this control, possibly through modulation of hypothalamic serotonin. The present work aimed at investigating the interaction of these systems. We addressed whether 5-HT is affected during IL 1beta-induced anorexia in obese Zucker rats and the influence of the central NO system on this IL-1beta/5-HT interaction. Using microdialysis, we observed that an intracerebroventricular injection of 10 ng IL-1beta significantly stimulated 5 HT extracellular levels in the VMH, with a peak variation of 102+/-41% above baseline. IL-1beta also significantly reduced the 4-h feeding by 33% and the 24-h feeding by 42%. Contrarily, these effects were absent when IL-1beta was injected 2 h after the i.c.v. administration of 20 microg of the NO precursor L-arginine. The results suggest that, in obese Zucker rats, activation of the serotonergic system in the medial hypothalamus participates in IL-1beta-induced anorexia. Since L-arginine, probably through NO stimulation, abolished both the anorexia and the serotonergic activation, it can be proposed that the NO system, either directly or indirectly, counteracts IL-1beta anorexia. The hypothalamic serotonergic system is likely to mediate this NO effect. PMID- 15544853 TI - Somatostatin down-regulates LFA-1 activation by modulating Rap1 expression in CD4+ and CD8+ T cells. AB - Leukocyte function-associated antigen-1 (LFA-1) is one of the integrins that are expressed on the leukocytes, and has been shown to play an important role in leukocyte trafficking. The adhesive activity of LFA-1 is governed partially by the Rap1. This study examined that the relationship between LFA-1 and Rap1 mRNA expressions by anti-CD3 and anti-CD3+SOM treatment in the CD4+ and CD8+ T cells. The LFA-1 mRNA expression levels following the anti-CD3 and anti-CD3+SOM treatment for 30 min was greater on the CD8+ T cells, and the LFA-1 expression of the CD8+ T cells with anti-CD+SOM treatment was affected more severely than that of the CD4+ T cells. The Rap1 mRNA expression patterns following anti-CD3 and anti-CD3+SOM stimulation in the CD4+ and CD8+ T cells were similar to the LFA-1 expression patterns, and the expression level following anti-CD3+SOM treatment was suppressed more significantly in the CD8+ T cells. These results suggest that the difference in the Rap1 expression level after stimulation might explain the differences in the LFA-1 expression level on the T cell subsets, and that the down-regulation of Rap1 expression following SOM treatment is closely related to the diminished LFA-1 expression. PMID- 15544854 TI - Specific enkephalin-degrading aminopeptidase activity in the HPT and HPO axes of rats with breast cancer induced by N-methyl nitrosourea. AB - State and function of breast depend on an endocrinological balance, the upsetting of which can be a factor favorable to the development of cancer. Enkephalins (ENK) have been considered as a particular form of adaptation to defense to the organism against neoplastic processes. However, ENK may modify the endocrine functions of glands such as the ovary or the thyroid through the hypothalamus pituitary axis, acting direct or indirectly as endocrine, paracrine or autocrine stimulatory growth factors. The present work analyses enkephalin-degrading tyrosyl aminopeptidase (EDA) activity in the hypothalamus-pituitary-thyroid (HPT) and hypothalamus-pituitary-ovary (HPO) axes in a rat model of breast cancer induced by N-methyl-nitrosourea (NMU) to state the relationship between ENK levels modification through EDA activity at different neuroendocrine levels and breast cancer. Results obtained show a decrease in EDA activity in hypothalamus, anterior and posterior pituitary, thyroid and ovary, suggesting increased levels of ENK in all these locations. These ENK may induce breast cancer cell growth and progression not only at breast level, but also acting at several neuroendocrine levels such as the HPT and HPO axes, inducing an unbalance of several other hormones, which could also facilitate the progression of cancer as an undesirable concomitant effect. PMID- 15544855 TI - Neuropeptide Y and its receptor subtypes specifically modulate rat peritoneal macrophage functions in vitro: counter regulation through Y1 and Y2/5 receptors. AB - It is well documented that neuropeptide Y (NPY) exerts a wide range of biological functions through at least five NPY Y receptor subtypes (Y1-Y5), but its immunological effects only recently came into focus. Using NPY family peptides and NPY-related receptor-specific peptides as well as Y1 and Y2 receptor antagonists, we have tested which NPY Y receptors are involved in NPY-induced modulation of rat peritoneal macrophage function in vitro. NPY and PYY increased oxidative burst in phorbol myristate acetate (PMA)-stimulated macrophages involving activation of protein kinase C (PKC), and decreased it in zymosan stimulated cells resembling inhibition of signaling pathways subsequent to binding of zymosan particles for the iC3b fragment receptor on macrophages. The combined treatment with NPY and NPY Y receptor antagonists revealed that NPY induced potentiation of oxidative burst in PMA-stimulated cells is mediated through Y1 and Y2 receptors, while NPY-induced suppression in zymosan-stimulated cells is mediated through Y2 receptors only. NPY-related peptides differently modulated macrophage function, confirming involvement of NPY Y2 receptor in both potentiation and suppression of oxidative burst in these cells. Additionally, it was shown that NPY Y5 receptor mediated suppression of oxidative burst in PMA- and zymosan-stimulated macrophages. Taken together, the present data reveal an NPY Y1 and Y2/Y5 receptor interaction in NPY-induced modulation of macrophage functions related to inflammation. PMID- 15544856 TI - An antimicrobial peptide from the skin secretions of the mountain chicken frog Leptodactylus fallax (Anura:Leptodactylidae). AB - A 25 amino-acid-residue, C-terminally alpha-amidated peptide with antimicrobial activity, which has been termed fallaxin, was isolated in high yield from the norepinephrine-stimulated skin secretions of the mountain chicken frog Leptodactylus fallax (Anura:Leptodactylidae). The amino acid sequence of the peptide (Gly-Val-Val-Asp-Ile-Leu-Lys-Gly-Ala-Ala-Lys-Asp-Ile-Ala-Gly-His-Leu-Ala Ser-Lys-Val-Met-Asn-Lys-Leu.NH2) shows structural similarity with members of the ranatuerin-2 family previously isolated from the skins of frogs of the genus Rana that are only distantly related to the Leptodactylidae. This observation is consistent with the hypothesis that many frog skin antimicrobial peptides are related evolutionarily, having arisen from multiple duplications of an ancestral gene that existed before the radiation of the different families. Fallaxin inhibited the growth of reference strains of Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Enterobacter cloacae, Klebsiella pneumoniae) but with relatively low potency (MIC> or =20 microM) and was inactive against the Gram-positive bacterium (Staphylococcus aureus) and the yeast Candida albicans. The hemolytic activity of fallaxin was very low (HC50>200 microM). A second peptide, comprising residues (1-22) of fallaxin, was also isolated from the skin secretions but this component was inactive against the microorganisms tested. PMID- 15544857 TI - Cortistatin radioligand binding in wild-type and somatostatin receptor-deficient mouse brain. AB - Cortistatin-14 (CST-14) is a recently discovered member of the somatostatin family of neuropeptides. It shares 11 of its 14 amino acids with somatostatin-14 (SRIF-14). In the present study, binding sites for cortistatin-14 in the mouse brain were examined and compared to those for somatostatin using iodinated cortistatin-14 and iodinated somatostatin-14. By in vitro receptor autoradiography, high densities of cortistatin-14 and somatostatin-14 specific binding sites were detected in the cortex, hippocampal formation, basolateral amygdala and medial habenula. Unlabeled 100 nM cortistatin-14 inhibited iodinated somatostatin-14 binding in the hippocampus, but not in the cortex or amygdaloid nuclei. In somatostatin receptor subtype-2 knock-out (KO) mice, autoradiographic iodinated somatostatin-14 binding was observed in the hippocampus and habenula but was removed in the cortex and amygdaloid nuclei, specific iodinated cortistatin-14 binding sites were found in the hippocampus, habenula and throughout the cortex. We conclude that the somatostatin receptor subtype-2 is responsible for somatostatin binding in cortical and amygdaloid regions and that cortistatin predominantly interacts with the same receptors as somatostatin. PMID- 15544858 TI - Increased plasma levels of adrenomedullin, a vasoactive peptide, in patients with end-stage pulmonary disease. AB - AIM: To study adrenomedullin (AM) plasma levels in patients with severe lung disease and to analyze the relationship between AM and heart changes, hemodynamics and blood gases. METHODS: Case control study of 56 patients (36 men, 20 women) with severe lung disease and 9 control subjects (7 men, 2 women). Patients with end-stage pulmonary disease, including chronic obstructive pulmonary disease (COPD, n=11), cystic fibrosis (CF, 26), idiopatic pulmonary fibrosis (ILD, n=9), and idiopatic pulmonary arterial hypertension (PAH, n=10), who were evaluated for lung trasplantation between January 1997 and September 2000, and nine patients who underwent lung surgery for a solitary benign nodule. AM plasma levels in pulmonary artery (mixed venous blood, vein) and aorta or femoral artery (arterial, art), art and vein blood gases, pulmonary hemodynamics, systemic hemodynamics, two-dimensional transthoracic echocardiography and echo Doppler study. RESULTS: Plasma AM (art and ven) levels were higher among patients' group compared to the controls (AMart p<0.02 and AMven p<0.04) for CF, ILD, PAH (AMart, pg ml(-1) Controls 13.7+/-3.6, COPD 22.8+/-6.2, CF 28.1+/-11.4, ILD 34.1+/-14.3, PAH 35.1+/-18.9; AMven, pg ml(-1) Controls 14.2+/-4.8, COPD 28.1+/-12.6, CF 31.7+/-14.1, ILD 38.7+/-16.5, PAH 40.1+/-4.4). We found with a trend towards higher concentration in ILD and PAH patients compared to COPD and CF but no statistical significant differences. Mixed-venous AM was higher than arterial AM in all groups resulting in AM uptake (AMPulmUp pg min(-1) Controls 4.8+/-22.6, COPD 21.1+/-44.9, CF 20.6+/-45.1, ILD 23.7+/-38.5, PAH 29.9+/-49.7). The univariate analysis showed a weak but significant correlation between AMart and mean systemic arterial pressure, heart rate, mean pulmonary arterial pressure and systemic vascular resistance. In the multivariate analysis, four variables emerged as independent factors of AMart including mean pulmonary arterial pressure, heart rate, mean systemic arterial pressure and left ventricular diastolic diameter (F=8.6, p<0.00001, r=0.60, r2=0.32). A similar weak correlation was apparent between AMven, systemic vascular resistance, and mean pulmonary arterial pressure. The results of multivariate analysis identify right atrial enlargement, mean right atrial pressure, heart rate and left ventricular dimensions as the only independent variables related to AMven (F=4.3, p<0.0004 r=0.56, r2=0.26). AM pulmonary uptake was significantly correlated with AMven (r=0.65), but not with hemodynamic, blood gas and echocardiographic variables. CONCLUSIONS: AM plasma levels are elevated in patients with severe lung disease in face of a preserved pulmonary uptake. These results suggest that the high AM plasma levels in patients with severe lung disease are not caused by a reduced pulmonary clearance, instead suggesting a systemic production. PMID- 15544859 TI - Inflammatory cells as source of tachykinin-induced mucus secretion in chronic bronchitis. AB - Substance P and neurokinin A are regulatory peptides of the tachykinin family that influence many aspects of human airway function in health and diseases such as bronchial asthma or chronic obstructive pulmonary disease (COPD). Tachykinin induced mucus secretion has been regarded as sensory nerve-dependent so far. We studied the distribution of tachykinin-mRNA and -peptide and its relation to NK-1 subtype-positive cells in human airway glands to assess if tachykinins may also be expressed in inflammatory cells. RT-PCR demonstrated the expression of tachykinin- and NK-1-mRNA in human airway tissues. In situ hybridisation resulted in preprotachykinin (PPT)-A mRNA-signal detection in inflammatory cells which were in close contact to myoepithelial cells of airway glands. NK-1 immunoreactivity was found in myoepithelial cells which were in direct contact to the PPT-A mRNA and tachykinin-positive cells. The present data directly demonstrate the presence of both PPT-A mRNA and tachykinin immunoreactivity in inflammatory airway cells which are in direct contact to NK-1 receptor positive glandular myoepithelium. Our findings indicate that besides neurally released tachykinins, also inflammatory cell-derived tachykinins may lead to glandular secretion via NK-1 receptor stimulation. This points to a major second source of these proinflammatory mediators in chronic inflammatory airway diseases such as COPD or asthma. PMID- 15544860 TI - Nociceptin/orphanin FQ prevents ethanol-induced gastric lesions in the rat. AB - Nociceptin/orphanin FQ (N/OFQ), the endogenous ligand of the NOP receptor, exerts a variety of effects on the gastrointestinal tract. The present study was aimed at evaluating the possible implication of N/OFQ in the maintenance of gastric mucosal integrity. N/OFQ was given either centrally or peripherally 30 min prior to intragastric administration (i.g.) of 1 ml/rat of ethanol (either 25% or 50%, v/v), which produces macroscopically visible gastric lesions. Intracerebroventricular (i.c.v.) injection of 2 microg/rat of N/OFQ significantly reduced lesions caused by 50% ethanol, while 1 microg/rat was enough to significantly reduce lesions caused by 25% ethanol. Intracerebroventricular injection of 5 microg/rat of the selective NOP receptor antagonist, UFP-101, completely reversed the protective effect of N/OFQ, 1 or 4 microg/rat against 25% or 50% ethanol, respectively. The intraperitoneal (i.p.) injection of N/OFQ produced a significant reduction of lesions induced by 50% ethanol, the peak effect being observed at 10 microg/kg. Intraperitoneal pretreatment with UFP-101, 120 microg/kg, completely abolished the protective effect of peripherally injected N/OFQ. Therefore, N/OFQ acts both centrally and peripherally as a protective agent against ethanol-induced gastric lesions, and its effect is mediated by NOP receptors. PMID- 15544861 TI - Food intake in rhesus monkeys following central administration of orexins. AB - Orexin A has been reported to stimulate food intake in rats while orexin B does not. The purpose of this study was to determine the role of orexin A or orexin B administration on food intake in adult, male rhesus monkeys. Food intake was measured at 2 and 8 h after the morning feeding following central injections of vehicle, orexin A (10, or 20 microg) or orexin B (10, 30, or 100 microg). When compared to vehicle injections, the 10 and 20 microg doses of orexin A decreased food intake at 2 h post-dose by 45% and 64%, respectively. Eight-hour food intake was decreased at only the 20 microg orexin A dose. Orexin B at all doses and time points did not alter food intake when compared to vehicle. These results indicate that orexin A exhibits anorectic activity while orexin B does not affect food intake in the rhesus monkey. PMID- 15544862 TI - Sustained inhibitory effect of Agouti Related Protein on the ACTH-induced cortisol production by bovine cultured adrenal cells. AB - The adrenal gland is the second tissue after hypothalamus exhibiting high expression level of Agouti Related Protein (Agrp) mRNA, which suggests that this peptide may control adrenal cell functions. However, its role in this tissue remained to be determined. In this report, we studied the effects of a long-term treatment (24 h) of cultured bovine adrenal cells by Agrp on the (Nle4, d-Phe7) alphaMSH (NDP-alphaMSH)- or ACTH-induced cortisol release. We showed that Agrp inhibited, in a dose-dependent manner, the 10(-9) M NDP-alphaMSH-induced cortisol production through its antagonistic properties towards MSH at the level of MC4-R. Surprisingly, we found that Agrp in the same conditions of cell treatment also induced a strong inhibition of the ACTH-induced cortisol release. These effects were stronger using low concentrations of Agrp and disappeared for higher concentrations resulting in U-shaped curve data. There was no effect of SHU9119 in the same conditions of stimulation of the cells. Our data confirmed that Agrp is not an antagonist of ACTH at the level of MC2-R and that its sustained effect on ACTH-induced steroidogenesis did not involve its antagonistic properties at the level of MC4-R. The hypothesis would be that Agrp is acting on adrenal steroidogenesis through an alternate mechanism. PMID- 15544863 TI - Enhanced dermal and retinal vascular permeability in streptozotocin-induced type 1 diabetes in Wistar rats: blockade with a selective bradykinin B1 receptor antagonist. AB - The vascular complications associated with type 1 diabetes are to some extent related to the dysfunction of the endothelium leading to an increased vascular permeability and plasma extravasation in the surrounding tissues. The various micro- and macro-vascular complications of diabetes develop over time, leading to nephropathy, retinopathy and neuropathy and cardiomyopathy. In the present study, the effect of a novel selective bradykinin B1 receptor (BKB1-R) antagonist, R 954, was investigated on the changes of vascular permeability in the skin and retina of streptozotocin (STZ)-induced type 1 diabetic rats. Plasma extravasation increased in the skin and retina of STZ-diabetic rats after 1 week and persisted over 4 weeks following STZ injection. Acute treatment with R-954 (2 mg/kg, bolus s.c.) highly reduced the elevated vascular permeability in both 1- and 4-week STZ diabetic rats. These results showed that the inducible BKB1-R subtype modulates the vascular permeability of the skin and retina of type 1 diabetic rats and suggests that BKB1-R antagonists could have a beneficial role in diabetic neuropathy and retinopathy. PMID- 15544865 TI - Cationic compounds used in lipoplexes and polyplexes for gene delivery. AB - Gene transfer represents an important advance in the treatment of both genetic and acquired diseases. Many cationic lipids and cationic polymers naturally occurred or synthesized have been used for gene transfer. They have the advantages over viral gene transfer as non-immunogenic, easy to produce and not oncogenic. These cationic compounds, however, have the major limitations of inefficient transfection and toxicity to cells. For overcoming these problems, many new cationic compounds were developed since the first cationic lipid, DOTMA, was found usage in gene therapy. This article reviews cationic lipids for gene therapy from chemistry viewpoint and we classify these compounds as monovalent cationic lipids, polyvalent cationic lipids, cationic polymers, guanidine containing compounds, cationic peptides and cholesterol containing compounds, and hope to provide suggestions on the development of this variety of cationic compounds through the discussion. PMID- 15544866 TI - Excised porcine skin experimental systems to validate quantitative microdialysis methods for determination of drugs in skin after topical application. AB - Microdialysis is useful as a method to evaluate the disposition of drugs in the skin to design improved transdermal delivery systems (TDDSs). In this study, quantitative microdialysis methods were validated in excised porcine skin experimental systems in vitro. Flurbiprofen (FP), used as a model drug, showed high affinity for the skin tissues in equilibrium states between the medium and skin. The membrane clearances of FP for permeation through the membrane of a dialysis fiber placed in the skin (CL(m in S)) were lower than that in the medium. The adsorption of components in the skin to the membrane surface of the dialysis fiber and accumulation of FP near the dialysis fiber are the most likely reasons for this. When CL(m in S) was used to predict the extracellular FP concentration in skin (C(T)), the value obtained was lower than that expected from the FP concentration in the medium on the dermis side, which should be equal to C(T) at equilibrium. In the zero net flux (ZNF) method, in which the concentration difference of perfusate (DeltaC) between the inflow and outflow were used to obtain C(T), the predicted C(T) was similar to the expected value. In an in vitro skin permeation experiment, the ZNF method was used for the prediction of C(T) near the dialysis fiber. The predicted C(T) was over 10 times higher than the FP concentration in the medium on the dermis side, suggesting a concentration gradient in the dermis. Although the ZNF method is good for predicting the C(T) in skin, the mass balance has to be considered for the quantitative evaluation of the skin permeation of drugs. In this study, the effect of the mass transfer of FP from the perfusate to the skin on the cumulative amount of FP passing through the skin was relatively low because of the use of suitable solutions as perfusate. The perfusion conditions and schedules should be designed carefully for quantitative evaluations using the ZNF method. These results provide useful information for the in vivo application of quantitative microdialysis to evaluate TDDS. PMID- 15544867 TI - Transdermal delivery from a lipid sponge phase--iontophoretic and passive transport in vitro of 5-aminolevulinic acid and its methyl ester. AB - The hydrochloride salts of 5-aminolevulinic acid (ALA) and its methyl ester (m ALA), respectively, were dissolved in a lipid sponge phase comprising monoolein, propylene glycol and aqueous buffer at concentrations of approximately 0.25% and 16% w/w m-ALA. The iontophoretic and passive delivery of ALA and m-ALA from this formulation through porcine skin in vitro were measured and compared to formulations used in clinical practice, 20% w/w ALA in Unguentum M and Metvix (a cream containing 16% w/w m-ALA). A sponge phase with 16% w/w m-ALA showed a higher passive flux (approximately 140 nmol cm(-2) h(-1) at 5 h) but a lower iontophoretic flux (approximately 800 nmol cm(-2) h(-1) at 5 h) compared to the clinically used products but the differences are hardly significant due to large standard deviations. ALA and m-ALA in sponge phase formulation showed iontophoretic fluxes in the range 80-100 nmol cm(-2) h(-1) at 3 h, i.e. values comparable to the passive fluxes from the more concentrated vehicles. The results demonstrate that the lipid sponge phase, a thermodynamically stable liquid with amphiphilic character, may have potential as a transdermal drug delivery vehicle. PMID- 15544868 TI - Transdermal self-permeation enhancement of ibuprofen. AB - The objective of this study was to prepare saturated solutions of ibuprofen, of different concentrations, and to investigate their effect on permeation of ibuprofen across rat epidermis. Ibuprofen saturated solutions were prepared using 0.1, 0.2, 0.3 and 0.4 M disodium hydrogen phosphate solution (DHP). The solubility of ibuprofen in DHP increased as the molarity of DHP increased. Thus the four saturated solutions of ibuprofen (0.1M-DHP-IBU, 0.2M-DHP-IBU, 0.3M-DHP IBU and 0.4M-DHP-IBU) have different concentrations of the same drug, and showed same pH (pH 7.0+/-1). The permeability study was also carried out using human epidermis and silastic membrane. Permeation rate of ibuprofen across rat epidermis and human epidermis from 0.4M-DHP-IBU was much greater than from 0.1M DHP-IBU. The magnitudes of increase in the drug flux were 46.4-fold with rat epidermis and 9.4-fold with human epidermis. Such a great increase in drug flux was not observed with silastic membrane, only 1.4-fold. This suggests that the increased drug flux is likely due to drug-skin interaction and not the increased concentration of ibuprofen per se. Surface tension (ST) measurements of DHP versus ibuprofen concentration showed ST reduction of DHP, from 72 to 27.9 dyn/cm. This is an indication that ibuprofen acted as ionic surfactant and the observed skin permeability enhancement is attributed to disruption of stratum corneum barrier. Results of DSC study supported this assumption. DSC of untreated rat stratum corneum samples showed lipid transitions at 41.9+/-0.0 degrees C (T1), 55.1+/-1.6 degrees C (T(x)), 70.2+/-0.1 degrees C (T2) and 77.5+/-0.1 degrees C (T3), while those pretreated with 0.4M-DHP-IBU did not show the first three lipid transitions. Also, pretreatment of rat epidermis with 0.4M-DHP-IBU enhanced permeation of diclofenac sodium greater than 1250-fold. This corroborates that ibuprofen not only enhances its own permeation but also that of other drugs, such as diclofenac sodium. PMID- 15544869 TI - In vivo release from a drug delivery MEMS device. AB - A drug delivery microelectromechanical systems (MEMS) device was designed to release complex profiles of multiple substances in order to maximize the effectiveness of drug therapies. The device is based on micro-reservoirs etched into a silicon substrate that contain individual doses of drug. Each dose is released by the electrochemical dissolution of the gold membrane that covers the reservoir. The first in vivo operation of this device was reported in this study. Subcutaneous release was demonstrated in rats using two tracer molecules, fluorescein dye and radiolabeled mannitol, and one radiolabeled chemotherapeutic agent, carmustine (BCNU). BCNU was chosen because of the need to improve the direct delivery of chemotherapy to malignant tumors. The spatial profile of fluorescein dye release from the drug delivery device was evaluated by fluorimetry, the temporal profile of 14C labeled mannitol release was evaluated by liquid scintillation counting, and the temporal profile of 14C labeled BCNU release was evaluated by accelerator mass spectrometry (AMS). Release profiles obtained from injected controls were compared with those from activated devices. The in vivo dye release results showed high concentration of fluorescein in the flank tissue surrounding the devices 1 h after activation. The 14C labeled mannitol released from the drug delivery devices was rapidly cleared (1 day) from the rat urine. In vivo release of 14C labeled BCNU from activated devices showed slightly slower kinetics than the injected and in vitro controls, and the time to reach the steady-state plasma 14C concentration was on the order of 1 h. All these results demonstrated the capability of this drug delivery device to achieve localized delivery of various compounds with well-defined temporal profiles. PMID- 15544870 TI - Skin absorption and metabolism of a new vitamin E prodrug, delta-tocopherol glucoside: in vitro evaluation in human skin models. AB - The aim of this study was to investigate the cutaneous penetration and metabolism of the new vitamin E prodrug delta-tocopherol glucoside (delta-TG), as compared to those of common vitamin E acetate, in vitro, both in reconstituted human epidermis and in viable human skin. Better diffusion was observed with alpha tocopherol acetate (alpha-TAc) than with delta-tocopherol glucoside in both skin models, at 0.1% and 0.05% in a myritol solution; however, no metabolism was detected with alpha-tocopherol acetate. In all conditions tested (two skin models, two concentrations, three test times, and compartmental analysis) the delta-tocopherol glucoside was metabolized into free tocopherol. In the reconstituted human epidermis, after 18 h, over 90% of the delta-tocopherol glucoside was bioconverted. In the viable human skin, the extent of metabolism was about 20%, with 0.12 and 0.10 microg/cm2 of delta-tocopherol glucoside in the stratum corneum and epidermis, respectively. After topical application, the delta tocopherol glucoside had a considerable reservoir effect, associated with gradual delivery of free tocopherol. The use of this gluco-conjugated vitamin E at a low concentration shows the capability of the skin to metabolize the prodrug in a slow and prolonged manner, making this gluco-conjugated vitamin E an excellent candidate for continuous reinforcement of antioxidants in the skin. PMID- 15544871 TI - Effect of Aerosil on the properties of lipid controlled release microparticles. AB - Theophylline-loaded microparticles of a lipid carrier, Precirol ATO 5, were prepared by the ultrasonic spray-congealing method. The goal of the work was to investigate the effect of different concentrations and kind of colloidal silicon dioxide (Aerosil 90, 200 and 300) on the microparticle characteristics (particle size, drug loading, morphology and kinetics of release). The results showed that the introduction of Aerosil improved the drug distribution in the different particle sizes and that the mean diameter of the microparticles decreased with the viscosity of the suspension to be nebulized, especially that with Aerosil 300. Whatever the microparticles formulation is, SEM and image analysis did not reveal any remarkable difference of the microparticle shape and surface area, suggesting that other parameters could influence the dissolution behaviour. Actually, the dissolution profiles of all the formulations appeared to be closely related to the physico-chemical properties of Aerosil, especially to its gelation properties, which are a function of its specific surface area. In particular, microparticles having high concentration of Aerosil 200 and 300 approached a zero order release kinetics, while Aerosil 90 microparticles followed a first order release kinetics. Therefore, the drug release rate is controlled by the extent and rate of water absorption/swelling of the Aerosil employed. Finally, DSC, HSM, XRD and FT-IR evidenced the permanence of the drug in its original state. PMID- 15544872 TI - Folate-receptor-targeted delivery of doxorubicin nano-aggregates stabilized by doxorubicin-PEG-folate conjugate. AB - For folate-receptor-targeted anti-cancer therapy, doxorubicin aggregates in a nano-scale size were produced employing doxorubicin-polyethylene glycol-folate (DOX-PEG-FOL) conjugate. Doxorubicin and folate were respectively conjugated to alpha- and omega-terminal end group of a PEG chain. The conjugates assisted to form doxorubicin nano-aggregates with an average size of 200 nm in diameter when combined with an excess amount of deprotonated doxorubicin in an aqueous phase. Hydrophobically deprotonated doxorubicin molecules were aggregated within the core, while the DOX-PEG-FOL conjugates stabilized the aggregates with exposing folate moieties on the surface. The doxorubicin nano-aggregates showed a greater extent of intracellular uptake against folate-receptor-positive cancer cells than folate-receptor-negative cells, indicating that the cellular uptake occurred via a folate-receptor-mediated endocytosis mechanism. They also exhibited more potent cytotoxic effect on KB cells than free doxorubicin. In a human tumor xenograft nude mouse model, folate-targeted doxorubicin nano-aggregates significantly reduced the tumor volume compared to non-targeted doxorubicin aggregates or free doxorubicin. These results suggested that folate-targeted doxorubicin nano aggregates could be a potentially useful delivery system for folate-receptor positive cancer cells. PMID- 15544873 TI - Controlled release of hyaluronan oligomers from biodegradable polymeric microparticle carriers. AB - In the present study, biodegradable microparticles of blends of poly(DL-lactic-co glycolic acid) (PLGA) and poly(ethylene glycol) (PEG) were explored as a potential carrier for the controlled release of polysaccharide oligomers. To this end, hyaluronan (HY) oligomers of varying molecular weights were incorporated into PLGA/PEG microparticles. Using a two-level fractional factorial experimental design, four microparticle formulation parameters, the amount of PEG included in the microparticles, the initial HY loading of the microparticles, the molecular weight of HY, and the molecular weight of PLGA, were studied for their influence on the incorporation and in vitro release of HY over the period of 28 days. The entrapment efficiencies were found to range between 10+/-1% and 24+/-2% depending on the initial loading and the molecular weight of the HY oligomer used in the fabrication of the microparticles. The HY was released in a multiphasic fashion including an initial burst release, followed by two separate periods of linear release. The normalized cumulative mass released during the burst release ranged from 25.1+/-9.2% to 93.0+/-0.7% and was found to be significantly influenced by the initial HY loading, the HY molecular weight, and the PLGA molecular weight. The initial period of linear release lasted from day 1 to day 14 and displayed normalized cumulative rates of release from 0.1+/-0.0%/day to 1.4+/-0.2%/day. During this period, PEG content of the microparticles and HY molecular weight exerted the greatest influence on the rate of release. Finally, the second period of linear release lasted through the final time-point at day 28. Here, the normalized cumulative rate of release values ranged from 0.2+/-0.1%/day to 3.6+/ 0.7%/day and were dependent on all formulation parameters studied. These results demonstrate the potential of PLGA/PEG blend microparticles for the controlled release of HY oligomers. PMID- 15544874 TI - A stochastic differential equation model for drug dissolution and its parameters. AB - A stochastic differential equation describing the process of drug dissolution is presented. This approach generalizes the classical deterministic first-order model. Instead of assuming a constant fractional dissolution rate, it is considered here that the rate is corrupted by a white noise. The half-dissolution time is investigated for the model. The maximum likelihood and Bayes methods for the estimation of the parameters of the model are developed. The method is illustrated on experimental data. As expected, due to the nonlinear relationship between the fractional dissolution rate and the dissolution time, the estimates of the dissolution rate obtained from this stochastic model are systematically lower than the rate calculated from the deterministic model. PMID- 15544875 TI - Tumor-targeted bioconjugate based delivery of camptothecin: design, synthesis and in vitro evaluation. AB - Camptothecin (CPT) presents numerous challenges associated with optimal transport and delivery including variability in clinically observed effects, low target tissue concentrations and severe and unpredictable toxicity. The objective of the present study was to optimize the delivery of CPT by targeting it to cancer cells using an endogenous receptor system. A novel CPT bioconjugate was synthesized using carbodiimide chemistry with a linear poly(ethylene glycol) (PEG) and amino acid glycine as the spacer and linker respectively. Folic acid was used as the targeting ligand to take advantage of folate receptor mediated endocytosis. The bioconjugate was extensively characterized using MALDI, proton NMR, FT-IR and amino acid analysis. Furthermore, the bioconjugate was evaluated in vitro for specific targeting to folate receptor-expressing KB cells, a human nasopharyngeal carcinoma. Finally, the delivery system was evaluated for cytotoxicity using a MTT based assay. The results indicate significantly higher efficacy of the bioconjugate in comparison to CPT. A control conjugate without PEG demonstrated no improvement in efficacy over untargeted CPT emphasizing the importance of spacer between the anticancer compounds and targeting moiety. This bioconjugate represents the 'first-in-series' of targeted bioconjugates and serves as prototype for improving tumor cell concentration and efficacy. PMID- 15544876 TI - Phosphorylcholine-polycation diblock copolymers as synthetic vectors for gene delivery. AB - A novel 2-(dimethylamino)ethyl methacrylate-block-2-(methacryloyloxyethyl phosphorylcholine) (DMAEMA-MPC) diblock copolymer was synthesized and investigated as a new non-viral vector for gene delivery. The attractive perspective of this phosphorylcholine (PC)-based material is its propensity to condense DNA efficiently via the cationic DMAEMA block, as previously demonstrated for the respective homopolymer, with the MPC block acting as a biocompatible steric stabilizer. Two series of DMAEMA-MPC diblock copolymers were synthesized for evaluation, varying independently and systematically either MPC or DMAEMA block length. Markedly different DNA-copolymer complexes were observed depending on the copolymer molecular composition. Certain polymeric structures led to formation of highly condensed, sterically stabilized DNA complexes of 120 140 nm diameter, while some resulted in partly condensed DNA-polymer complexes with 'spaghetti' structures, indicating the importance of a copolymer composition to balance condensing and steric stabilization effect. A low level of non specific cellular association of the complexes with optimized physicochemical properties was seen, indicating the role of MPC surface layer in the interactions with biological membranes and important property in preventing promiscuous interactions with tissues in the body and potentially allowing for cellular specific delivery of the condensates following the attachment of a targeting ligand. PMID- 15544877 TI - Management of the neck in patients with T1 and T2 cancer in the mouth. AB - Our aim in this retrospective study was to evaluate the extent of control of metastatic disease in the neck and the survival of patients with T1 and T2 oral cancer. METHODS: All 171 patients with T1 and T2 squamous cell carcinoma (SCC) in the mouth were identified from our computerised database. All had had primary tumours resected and 21 patients with palpable neck nodes had therapeutic neck dissections. Among 150 patients with no palpable nodes, 75 had elective neck dissections, and 75 were observed. The decision to do an elective neck dissection was based on clinical criteria and was not randomised. RESULTS: Cervical nodes contained metastases in 17 of 21 patients who had therapeutic, and 27 of 75 who had elective, neck dissections. Neck metastases developed subsequently in 15 of 75 patients in the observed group, and 9 of these were salvaged by therapeutic neck dissection. The 5-year disease free survival was 19/21 after therapeutic dissection, 72/75 after elective dissection, and 69/75 in the observed group. Patients with cervical nodal metastases had significantly reduced 5-year survival compared with those without (63% and 91%, P = 0.003). PMID- 15544878 TI - Midfacial fractures in children and adolescents: a review of 492 cases. AB - We studied the records of children and adolescents admitted to our hospital with facial fractures between 1993 and 2002. During the 10-year period, 492 patients with 555 midfacial fractures were treated. Their ages ranged from 1 to 18 years, the peak incidence being between 16 and 18. Three-hundred and eighty (77%) were boys. The zygoma was the most commonly fractured bone (n = 286), and the hard palate the least commonly fractured (n = 6). Motor-vehicles were responsible for 272 (55%) of all fractures. Most fractures were treated by closed reduction, and only 139 (25%) were treated by observation. Complications, including unsatisfactory fracture repair and infection, were recorded in 18 (4%) and the overall mortality was 1% (n = 8). PMID- 15544879 TI - Antibiotic prophylaxis for orthognathic surgery: a prospective, randomised clinical trial. AB - A prospective, randomised, placebo-controlled double-blind clinical trial, compared short-term (1 day) and long-term (5 days) antibiotic prophylaxis after orthognathic surgery. Thirty four patients had single jaw or bimaxillary osteotomies and were given two perioperative doses of amoxycillin. Patients were then randomised to receive either placebo or amoxycillin for 5 days in a double blind manner. Postoperatively the patients were monitored for infection by scoring a series of validated measurements of infection. In the postoperative period four patients required additional antibiotics in the short-term group and two in the long-term group (P = 0.67). Morbidity scores were higher in the short term group, at 406 to 264 (P = 0.04), and when individual variables were compared there was a significant difference in the degree of swelling (P = 0.04). Although a 5-day regimen of antibiotic prophylaxis in orthognathic surgery did not decrease the incidence of postoperative infection significantly, it may decrease the morbidity of the operation. PMID- 15544880 TI - Advanced surface-recording techniques for computer-assisted oral and maxillofacial surgery. AB - Markerless recording of patients based on natural anatomical surfaces makes planning of computer-assisted surgery much easier, as it is not necessary to place and measure markers. Recording of the surgical site with a laser scan takes the place of conventional marker-based recording. We have used auricles as well as the maxilla and mandible as reproducible surfaces. The geometric congruence of the laser scanned surface with the corresponding surface in the computed tomographs data-set and the applied intraoperative accuracy after recording with a laser scanner have been evaluated, and the system was successful in the maxilla (mean precision: 0.8mm, standard deviation: 0.3mm). In the mandible, the tongue and mobile floor of the mouth led to geometric incongruence and inadequate laser scanning. An exact recording using auricles was possible only as long as the auricles had not been temporarily deformed by the head support during CT imaging. PMID- 15544881 TI - Outcome of treatment of Class II malocclusion by intraoral mandibular distraction. AB - Our aim was to find out long-term results of treatment in patients treated orthodontically and by mandibular distraction osteogenesis. Data on duration of treatment, costs, and results of 26 patients (13 girls and 13 boys) with a mean age of 15 years were analysed. The preoperative cephalograms were compared with those taken at the last follow-up visit. There was a significant reduction in duration of treatment when patients were treated without a first phase that included functional appliances. The differences in costs of orthodontic treatment were not significant. The costs of the operation for distraction were significantly higher compared to BSSO, mainly because of the costs of the distraction devices. Comparison of the cephalograms showed a significant increase in SNB angle, Wits value, ANB angle, overjet, and overbite. The Y-axis, MP/S, and SpP/MP angle increased. Orthodontic treatment and distraction of the mandible was a successful, but more expensive, treatment. PMID- 15544882 TI - Intraoral distraction osteogenesis to lengthen the ascending ramus. Experience with seven patients. AB - Seven children with facial asymmetry, mean age 12 years (range 11-14.5) were treated by intraoral distraction osteogenesis to lengthen the hypoplastic ramus. We achieved a mean increase in length of the ramus of 13mm (range 10-16). In only one patient did we achieve a posterior open bite on the distraction side. All patients ended with a symmetrical chin. It was helpful to place an orthodontic bite block on the opposite side either preoperatively or postoperatively to cant the plane of occlusion. The duration of follow-up was too short to allow conclusions to be drawn about the future requirement for bimaxillary osteotomies. PMID- 15544883 TI - Marrow-derived osteoblasts seeded into porous natural coral to prefabricate a vascularised bone graft in the shape of a human mandibular ramus: experimental study in rabbits. AB - To find out if it was possible to prefabricate bone graft in the shape of a human mandibular ramus that possessed a pedicle that carried blood. The pore size of the natural coral was about 200 microm with a porosity of about 36%. The natural coral was made into the shape of a human mandibular ramus. Marrow-derived osteoblasts were seeded into porous natural coral scaffolds in a density of 2 x 10(8)/mL in 300 microL cell suspension. After two days of in vitro incubation, five cell-coral complexes were implanted into cell donor rabbits under the inferior epigastric blood vessels to prefabricate a vascularised bone graft of specific shape. Two months later the bone formation was observed by gross inspection and histological examination. Two months after operation, a well vascularised bone graft in the shape of the initial coral scaffold and with a blood-carrying pedicle had been regenerated successfully. Osteogenesis followed the pattern of endochondral bone formation. New bone could be seen on the surface and in the pores of coral on histological examination. We have shown that it is possible to fabricate vascularised bone graft in a predetermined shape using tissue engineering. This kind of bone graft may have future clinical application. PMID- 15544884 TI - Glandular function after intraoral removal of salivary calculi from the hilum of the submandibular gland. AB - We studied 43 patients (25 women and 18 men) who had salivary calculi removed from the hilum of the submandibular gland. Preoperatively they had clinical and radiographic examinations, and glandular function was measured scintigraphically in 38 patients. Postoperative follow-up was based on history, clinical examination, structured questionnaire, and scintigraphy. Stone(s) were removed successfully in 42 patients (97%). During the follow-up of a mean of 24 months (range 4-47), 37 patients were symptom-free and 2 patients had mild obstructive symptoms that did not require intervention. The other 4 patients had repeated infections that necessitated removal of the gland under general anaesthesia. Preoperative and postoperative scintigraphic assessments were made in 37 patients (88%). There was a significant increase in the functional fraction and the excretion rate in the gland after removal of the calculus. We conclude that glandular function improves to varying degrees in most patients after the removal of a salivary calculus. PMID- 15544885 TI - DNA microarray technology: insights for oral and maxillofacial surgeons. AB - Since the discovery of DNA and the recent sequencing of the entire human genome, there have been great advances in our understanding of the genetic basis of many diseases, including cancer. An exciting recent development is DNA microarray technology. This technique allows many genes to be studied in the same experiment rather than one gene at a time. It can therefore provide large amounts of data much more quickly than was previously possible. This has great implications for diseases such as cancer, which often show genetic damage in many parts of the genome. DNA microarray technology has now been used in the investigation of many tumours including melanoma, breast cancer, and lymphoma, as well as in the understanding of the genetic basis of metabolic diseases. However, it seems that our specialty knows little about the technique and its possible clinical applications. We give here a simple introduction to the technology and its likely role in the future management of oral cancer. PMID- 15544886 TI - Conservative treatment of large cystic lesions of the mandible: a prospective study of the effect of decompression. AB - Our aim was to evaluate prospectively the effects of decompression as the primary treatment of large mandibular cysts, irrespective of their histological type. Twenty patients with large mandibular cysts completed treatment successfully. After a mean duration of decompression of 446 days, cysts had shrunk by a mean of 81%. Mean follow-up was 527 days after removal of the decompression stent and removal of the cyst. There were no recurrences. The advantages of this approach are simplicity, immediate gathering of information on the type of cyst with simultaneous start of treatment, low morbidity and low incidence of complications during treatment. Despite the length of the treatment, we recommend the use of decompression stents in the treatment of large mandibular cysts. PMID- 15544887 TI - Anxiety and extraction of third molars in Turkish patients. AB - Preoperative anxiety is widespread and adversely affects a patient's physical and psychological outcome. Extraction of third molars is common, and many patients complain of anxiety and emotional disturbance. We assessed the anxiety of patients in Turkey before extraction of third molars. A total of 120 patients were admitted for removal of one or more third molars under local anaesthesia. The Amsterdam Preoperative Anxiety and Information Scale (APAIS) and Spielberger's State Trait Anxiety Inventory (STAI) were used to evaluate anxiety. The results showed that women were significantly more anxious than men; women who had not had a previous operation were more anxious than other women; there was no difference in the anxiety scores of patients who had previously had a local anaesthetic and those who had not; there were no differences in anxiety as measured by trait scores; patients who wanted a lot of information were more anxious. PMID- 15544888 TI - Metronidazole for the prevention of dry socket after removal of partially impacted mandibular third molar: a randomised controlled trial. AB - We randomised 119 patients who had been referred for removal of partially impacted mandibular third molars to be given either metronidazole 1600 mg or placebo as a single dose 45 min before operation. Ten of the fifty-nine patients who were given metronidazole and 13 of the 60 given placebo developed dry sockets. Two variables were significantly associated with the development of a dry socket: pericoronitis and oral contraceptives. PMID- 15544889 TI - Histological changes in the temporomandibular joint in rabbits depending on the extent of mandibular lengthening by osteodistraction. AB - We studied the histological changes in the temporomandibular joint (TMJ) after unilateral mandibular distraction osteogenesis in rabbits. Eight rabbits were used, two of which served as controls and the other six had distraction of the left mandibular body after a latency period of 7 days at a rate of 0.5mm a day for a total of 2mm (n = 2), 3.5mm (n = 2), and 5mm (n = 2) of distraction. After a 14-day consolidation period, TMJs from both sides were harvested and prepared for histological examination under an optical microscope using haematoxylin and eosin stain. We found no degenerative or inflammatory changes in either TMJ in any of the groups. Endochondral ossification in the condyle was greater on the opposite side in the experimental group than in the condyles of the control group. Endochondral ossification was active in the 3.5-mm group. PMID- 15544890 TI - Opinions of UK specialists about terminology, diagnosis, and treatment of atypical facial pain: a survey. AB - There are few robust, evidence-based data about what constitutes the diagnosis of atypical facial pain and how it is best treated. We therefore aimed to find out the current opinion of those on specialist lists in the United Kingdom (UK) on whether they use the term atypical facial pain, how they reach their diagnosis, and what treatment they offer. We sent out questionnaires to 240 specialists randomly selected from the UK lists of those most likely to deal with atypical facial pain (oral and maxillofacial surgeons, oral medical specialists, ear nose and throat surgeons, anaesthetists, psychiatrists and neurologists). We divided the replies according to whether the specialists were medically or dentally based. Of the 209 valid questionnaires, 143 were returned (a response rate of 68%); 127 of the 143 used the term atypical facial pain, the others used various other terms. The two groups used significantly different criteria, mainly to exclude other conditions to achieve a diagnosis. About half used haematological tests and most used radiographic investigations routinely, and there were no significant differences among the specialities. No unified pattern of referral between different units was found. Treatment was mainly by antidepressant and anticonvulsant drugs, and counselling. PMID- 15544891 TI - Transient femoral neuropathy after harvest of bone from the iliac crest. AB - A 31-year-old woman had bone harvested from the left anterior iliac crest as a graft for an augmentation genioplasty. For postoperative analgesia, she was given a bupivacaine infusion into the iliac wound. She developed a temporary left femoral mononeuropathy from which she recovered completely. PMID- 15544892 TI - Protection of the lip from the activator arm of maxillary distractors. AB - The long spring-loaded distractor arms of maxillary distraction devices can cause pain and ulceration of the upper lip. We describe a simple method to relieve the tension and discomfort on the soft tissues using Silastic tubing. PMID- 15544893 TI - Simple modified preoperative nasoalveolar moulding in infants with unilateral cleft lip and palate. AB - This paper reports a new preoperative nasoalveolar molding (PNAM) for infants with unilateral cleft lip and palate. The nasal stent of this PNAM is made of cobalt-chrome wire (1mm in diameter). A loop about 3mm in diameter in the middle of the stent enables precise manual control of the force and direction of the stent with considerable ease. PMID- 15544894 TI - Plate and screw "medallion" in the maxillofacial clinic. AB - Demonstration of the plates and screws used in the management of facial trauma and orthognathic surgery patients is made considerably easier with the use of a simple "medallion" containing appropriate examples of each. PMID- 15544895 TI - Intraosseous vascular anomaly (haemangioma) of the zygoma. AB - Intraosseous vascular anomalies are rare, and although they have been described in the facial skeleton, those occurring in the zygomatic bone are extremely rare. We report the case of a 38-year-old woman who presented with a painless hard swelling in the left zygomatic prominence, which was diagnosed as a cavernous haemangioma after a bone biopsy. The further management of this lesion in discussed, with particular reference to surgical resection and reconstruction. PMID- 15544896 TI - Protocols for follow up of malignant salivary tumours. PMID- 15544897 TI - Multi-slice CT angiography in evaluation of extracranial-intracranial bypass. AB - Multi-slice CT (MSCT) scanners provided significant improvement in vascular applications. In this study, our purpose was to determine the clinical utility of MSCTA in evaluation of extracranial-intracranial (EC-IC) bypass patency. Eleven (4 men and 7 women; mean age, 46 years; age range, 19-68 years) consecutive patients who underwent MSCTA and DSA after EC-IC bypass surgery were evaluated retrospectively. All patients underwent DSA within 3 weeks of MSCTA. The indications for EC-IC bypass were severe stenosis or occlusion of intracranial arteries in seven patients and therapeutic occlusion of intracranial artery for unclippable giant aneurysm in four patients. Ten patients underwent superficial temporal artery (STA)-middle cerebral artery (MCA) bypass and one patient underwent occipital artery-posterior cerebral artery (PCA) bypass. Eight STA-MCA bypasses in six patients were patent on MSCTA which were confirmed on DSA. Two STA-MCA bypasses and one occipital artery-posterior cerebral artery (PCA) bypass were occluded in three patients on MSCTA and DSA. In one patient, minimal stenosis of the STA-MCA bypass was identified on MSCTA and DSA. In 1 patient, STA MCA bypass was not well seen on MSCTA and suspected for occlusion. DSA identified the occlusion of bypass on this patient. MSCTA seems to be a very promising noninvasive technique in evaluation of EC-IC bypass. PMID- 15544898 TI - ROI measurement of the signal intensity of precentral cortex in the normal brain. AB - OBJECTIVE: It has recently been described that perirolandic cortex generally had a low signal intensity (SI) in neurologically normal brain. The aim of this study was to confirm this finding by an objective quantitative study. MATERIALS AND METHODS: Turbo fluid attenuated inversion recovery (FLAIR) magnetic resonance (MR) images of 24 neurologically normal patients were evaluated retrospectively. Signal intensity measurements of the precentral and superior frontal cortices (SFCs) were obtained at a manually traced irregular region-of-interest (ROI). t Test for paired samples was used to evaluate the significance of differences between signal intensity measurements. RESULTS: Mean signal intensities of precentral and superior frontal cortices were 349.5 and 380.7, respectively, on the right, and 351.7 and 374.1 on the left hemisphere. The difference between the mean signal intensities of the side-matched precentral and superior frontal cortices was statistically significant (P < 0.001). CONCLUSION: Low signal intensity of the precentral cortex (PCC) in normal brain on turbo FLAIR images is an objective finding, confirmed by ROI measurement. PMID- 15544899 TI - Comparison of metric analysis of spinal structures, exemplarily of the ligamentum flavum, obtained with CT and MRI. AB - PURPOSE: Comparison of metric analysis of spinal structures, exemplarily of the ligamentum flavum, obtained with computed tomography (CT) (soft tissue window and bone window) and magnetic resonance imaging (MRI) (T1 and T2 weighted images). MATERIAL AND METHODS: Forty-six lumbar ligamenta flava of 46 patients (25 women and 21 men) were examined at a Somatom Plus 4 (Siemens, Erlangen, FRG) and at a 1.5 T clinical scanner (Magnetom Vision, Siemens, Erlangen, FRG). Two independent neuroradiologists measured the thickness of the ligamenta flava in mm. Statistics included Pearson's correlation coefficient and the intra-class correlation coefficient. RESULTS: Mean values did not differ significantly. The correlation coefficients varied between 0.69 and 0.98. The best correlation occurred comparing the same techniques in different windowing and weighting (CT: r = 0.98; MRI: r = 0.95). Correlating different techniques the combination of CT bone window and T1 weighted images presented the best result (r = 0.75). CONCLUSIONS: Because of the excellent correlation between the examined techniques CT as well as MRI can equally be used to measure distances of spinal structures. PMID- 15544900 TI - Imaging of peripheral nerve sheath tumors with pathologic correlation: pictorial review. AB - Peripheral neurogenic tumors include neurilemoma, neurinoma, and malignant peripheral nerve sheath tumors. All neurogenic tumors share common imaging features. Although differentiation between them is difficult, neurogenic origin can be suggested from their imaging appearances, including fusiform shape, relation to the nerve, "split-fat" sign, associated muscle atrophy and intrinsic imaging characteristics including "target sign" as well as from lesion location along a typical nerve distribution. Our purpose is to make an overview of imaging findings of each type of peripheral nerve sheath tumor with emphasis on characteristic signs and correlate with histologic features. Morton's neuroma and intraneural ganglion are also included as tumors of nerve origin. PMID- 15544901 TI - The value of MR angiography techniques in the detection of head and neck paragangliomas. AB - OBJECTIVE: The objective of this study was to compare three-dimensional phase contrast angiography (3D PCA), 2D time-of-flight (2D TOF), and 3D TOF magnetic resonance (MR) angiography and a proton density weighted technique in terms of their ability to detect head and neck paragangliomas. MATERIALS AND METHODS: 14 patients with 29 paragangliomas were examined at 1.5 T. Three MR angiography sequences (3D PCA, 2D TOF, and multi-slab 3D TOF) and a proton density (PD) weighted sequence were reviewed by four neuroradiologists. The gold standard was digital subtraction angiography. Presence of tumor was assessed in five grades of confidence. Sensitivity and specificity were calculated after dichotomizing the results. Data was analyzed using the logistic regression method. RESULTS: Mean sensitivity and specificity for the four observers were for PD: 72%/97%, for 3D PCA: 75%/90%, for 2D TOF: 66%/93%, and for 3D TOF: 90%/92%. Sensitivity was significantly better for 3D TOF MRA (P < 0.001). No substantial between-observer variation for tumor detection was present. CONCLUSION: Our results demonstrate that, using 3D TOF MRA, paragangliomas in the head and neck region can be detected with high sensitivity and specificity. Further investigation is necessary to judge the value of 3D TOF MR angiography against fat suppressed contrast enhanced T1 weighted and fat suppressed T2 weighted MR sequences to find the optimal imaging sequence for paragangliomas. PMID- 15544902 TI - Intraoral sonographic features of tongue cancer after radical radiotherapy. AB - PURPOSE: The purpose was to illustrate the sonographic changes of tongue cancer after radical radiotherapy. MATERIALS AND METHODS: In 24 patients with tongue cancer treated by interstitial brachytherapy (BRT) (uneventful recovery n = 16 , recurrence n = 5, severe soft tissue complication n = 3), follow-up examination were performed and both the margin and the vascular pattern were retrospectively assessed. We basically performed US examination at least once every 3 months after BRT within 1 year during follow-up period. RESULTS: In the healing process, intraoral sonography shows an unclear margin immediately after brachytherapy and a transient increase of the vascularity lasted within 6 months after BRT, followed by a decrease in the vascularity. The large difference in echogenicity between the primary site and the surrounding tissue and the increased difference suggested the possibility of either radiation ulceration or recurrence. CONCLUSION: Intraoral sonography could depict the sequence changes of the tongue after BRT, and it was thus useful to confirm the clinical findings of either radiation ulcers or recurrence. PMID- 15544903 TI - Superficial inflammatory and primary neoplastic lymphadenopathy: diagnostic accuracy of power-doppler sonography. AB - OBJECTIVE: To evaluate the sensitivity, specificity and diagnostic accuracy of a cut-off of the resistive index of 0.5 for the differentiation between inflammatory and neoplastic primary lymphadenopathies. SUBJECTS AND METHODS: We measured the resistive index of superficial enlarged lymph nodes in a total of 50 patients (29 males and 21 females; age range 12-72 years, mean age 41.6 year) using an ATL 5000 HDI. A resistive index greater than or equal to 0.5 indicated an inflammatory lymph node and a resistive index <0.5 was consistent with neoplastic primary lymphadenopathies. The gold standard was either surgical biopsy or lymph-node reduction seen with ultrasound examination after antibiotic therapy. RESULTS: The sensitivity of the resistive index for distinguishing inflammatory from neoplastic lymphadenopathy was 84.6%, the specificity 100% and the diagnostic accuracy 95.7% (P < 0.001, statistically significant). CONCLUSION: The results of this study indicate that power-Doppler using a resistive index cut off of 0.5 was a valid technique for distinguishing between inflammatory and primary neoplastic lymph nodes in patients with superficial lymphadenopathies. PMID- 15544904 TI - Comparison of CT and MR imaging for the detection of extranodal neoplastic spread in metastatic neck nodes. AB - PURPOSE: To compare computed tomography (CT) and magnetic resonance (MR) imaging for the detection of extranodal neoplastic spread (ENS) in metastatic cervical nodes from head and neck squamous cell carcinoma. MATERIALS AND METHODS: 17 patients with a squamous cell carcinoma of the head and neck underwent CT and MR imaging. The neck nodes were assessed for ENS and the results compared using pathology from the surgical resection. RESULTS: Radiologic-pathologic correlation was performed in 51 malignant nodes. The accuracy, sensitivity and specificity were respectively 73, 65, 93% for CT, and 80, 78, 86% for MR imaging. Comparison of CT and MR imaging showed that there was no significant difference between the two modalities for either sensitivity (P = 0.1317) or specificity (P = 0.3173). CONCLUSION: CT and MR imaging are comparable for the detection of ENS. PMID- 15544905 TI - The way to a man's heart is through his stomach: much 'diaphragmatic' attenuation is likely gastric, and effervescent granules enhance cardiac imaging. AB - Avoidance of falsely positive results depends on distinguishing reality from artifact, in turn depending on images of highest quality. In radionuclide cardiac imaging, an inferior wall artifactual defect, so called "diaphragmatic attenuation", is particularly common and vexing. Despite the historically held view, analysis and review of the literature suggest the defect is likely not diaphragmatic but rather primarily due to attenuation by nearby stomach wall. The explanation is based on gravity and anatomy. With this improved understanding, effervescent granules were given as a clinical, nonresearch measure to nine patients during myocardial scanning. It was observed that two-thirds demonstrated moderate or marked lessening of attenuation. An additional benefit is lessening of artifact by extracardiac activity. These benefits may also apply to other sorts of cardiac radionuclide imaging. The significance of this new imaging method is discussed and various avenues of research are proposed. PMID- 15544906 TI - A mammographic image analysis method to detect and measure changes in breast density. AB - We present an image analysis method that can detect and measure breast density from digitised mammograms. We present initial results on applying our method to characterise breast changes, in particular, changes due to Hormone Replacement Therapy (HRT). It has been established that long-term use of certain hormone replacement therapies can increase the risk of breast cancer, a fact that encourages the notion that objective measures of tissue density can be an important development in breast cancer image analysis. A set of 59 temporal pairs of mammograms of patients undergoing HRT (two images per patient) were used. The clinician's assessment of density changes constituted the ground truth for evaluating the proposed quantitative measures of density change. The measures we developed are based on the Standard Mammogram Form (SMF) representation of interesting tissue and their performance (agreement with the expert's description) is also compared to the "interactive thresholding" method that has been used in the past to characterise mammographic density. The results clearly indicate that present methods for measuring mammographic density fail to characterise temporal changes while the proposed measures have the potential to aid the radiologist in assessing temporal density changes both on a global and a local basis. PMID- 15544907 TI - Ethanol sclerotherapy of peripheral venous malformations. AB - BACKGROUND: Venous malformations are congenital lesions that can cause pain, decreased range of movement, compression on adjacent structures, bleeding, consumptive coagulopathy and cosmetic deformity. Sclerotherapy alone or combined with surgical excision is the accepted treatment in symptomatic malformations after failed treatment attempts with tailored compression garments. OBJECTIVES: To report our experience with percutaneous sclerotherapy of peripheral venous malformations with ethanol 96%. PATIENTS AND METHODS: 41 sclerotherapy sessions were performed on 21 patients, aged 4-46 years, 15 females and 6 males. Fourteen patients were treated for painful extremity lesions, while five others with face and neck lesions and two with giant chest malformations had treatment for esthetic reasons. All patients had a pre-procedure magnetic resonance imaging (MRI) study. In all patients, 96% ethanol was used as the sclerosant by direct injection using general anesthesia. A minimum of 1-year clinical follow-up was performed. Follow-up imaging studies were performed if clinically indicated. RESULTS: 17 patients showed complete or partial symptomatic improvement after one to nine therapeutic sessions. Four patients with lower extremity lesions continue to suffer from pain and they are considered as a treatment failure. Complications were encountered in five patients, including acute pulmonary hypertension with cardiovascular collapse, pulmonary embolus, skin ulcers (two) and skin blisters. All patients fully recovered. CONCLUSION: Sclerotherapy with 96% ethanol for venous malformations was found to be effective for symptomatic improvement, but serious complications can occur. PMID- 15544908 TI - Internal jugular vein access for the interventional management of nonfunctioning artero-venous haemodialysis fistulas. AB - We report our experience and results with the use of internal jugular vein as secondary approach for the endovascular treatment of twelve nonfunctioning artero venous (AV) fistulas in haemodialysis patients. PMID- 15544909 TI - Impact of short-term hemodialysis catheters on the central veins: a catheter venographic study. AB - OBJECTIVE: To determine the incidence of pericatheter sleeve formation, thrombus formation, and stenosis of the central veins in hemodialysis patients with temporary catheters. METHODS AND MATERIAL: In this prospective study, 57 patients (40 males, 17 females) with temporary dialysis catheters had catheter venography by pulling back the catheter just before removal. Patient's age range was 25-87 years (mean age, 51 years). The venographic studies were evaluated for pericatheter sleeve formation, thrombus formation, and stenosis of the brachiocephalic vein (BCV) and the superior vena cava (SVC). The IJV could only be evaluated if there was adequate filling during contrast administration. In a subgroup of patients who had had only right IJV or only right SCV catheters, impact of these catheters on the central veins was compared. RESULTS: The catheter location was right internal jugular vein (IJV) in 26 cases, right subclavian vein (SCV) in 27 cases, left IJV in 1 case, and left SCV in 3 cases. Thirty-two patients (56%) had had only one temporary catheter and the rest had had more than one inserted. The mean dwell time for the catheters was 21 days (range 7-59 days). A pericatheter sleeve was detected on venography in 32 (56%) patients and thrombus formation was noted in 16 patients (28%). A total of 41 patients (72%) exhibited pericatheter sleeve and/or thrombus formation. While 19 of the 32 patients (59%) without previous catheterization had a sleeve around the catheter, only 13 (52%) of 25 patients who had had multiple catheters inserted had a sleeve (P > 0.005). Of the eight patients (14%) with BCV stenosis, two had >50% stenosis. Only one patient (2%) had mild stenosis of the SVC. Three patients out of 15 (20%) who had diagnostic venography for the IJV had severe stenosis of the vein. Pericatheter sleeve formation was more frequent in women (P < 0.005). However, there were no statistical differences with respect to pericatheter sleeve formation, luminal filling defect and BCV stenosis when patients were grouped according to age, dwell time of the catheter, number of catheters inserted, and diameter of the SVC. Forty-two of the fifty-seven patients had had only right IJV (n =16) or right SCV (n = 26) catheters. There were no differences between these groups with respect to rates of pericatheter sleeve formation, thrombus formation, or BCV stenosis. CONCLUSION: This study showed that even short-term catheters result in significantly high rates of pericatheter sleeve and thrombus formation which are two of the important causes of catheter malfunction. The IJV route is known to be much safer than the SCV route with respect to stenosis formation in the vein in which the catheter is inserted; however, the result showed no differences between the two routes with respect to frequencies of pericatheter sleeve formation, thrombus formation, and BCV stenosis. These findings remind us again that we should avoid unnecessary catheter insertion even for short-term in these chronically ill patients. PMID- 15544910 TI - Tuberculous arthritis of the appendicular skeleton: MR imaging appearances. AB - Tuberculosis [TB] of the appendicular skeleton is an uncommon infection caused by the tuberculous bacilli and constitutes only 1-3% of all tuberculosis infections. MR imaging features of tuberculous arthritis include bone marrow oedema, cortical erosions, synovitis, joint effusion, tenosynovitis, soft tissue collections, and myositis. These imaging features are at times non-specific, but in the correct clinical context help in diagnosis of tuberculosis. We present the various pathological manifestations of TB arthritis involving the different joints of appendicular skeleton and discuss their MR imaging appearances. PMID- 15544911 TI - Definition of pertinent parameters for the evaluation of articular cartilage repair tissue with high-resolution magnetic resonance imaging. AB - To evaluate articular cartilage repair tissue after biological cartilage repair, we propose a new technique of non-invasive, high-resolution magnetic resonance imaging (MRI) and define a new classification system. For the definition of pertinent variables the repair tissue of 45 patients treated with three different techniques for cartilage repair (microfracture, autologous osteochondral transplantation, and autologous chondrocyte transplantation) was analyzed 6 and 12 months after the procedure. High-resolution imaging was obtained with a surface phased array coil placed over the knee compartment of interest and adapted sequences were used on a 1 T MRI scanner. The analysis of the repair tissue included the definition and rating of nine pertinent variables: the degree of filling of the defect, the integration to the border zone, the description of the surface and structure, the signal intensity, the status of the subchondral lamina and subchondral bone, the appearance of adhesions and the presence of synovitis. High-resolution MRI, using a surface phased array coil and specific sequences, can be used on every standard 1 or 1.5 T MRI scanner according to the in-house standard protocols for knee imaging in patients who have had cartilage repair procedures without substantially prolonging the total imaging time. The new classification and grading system allows a subtle description and suitable assessment of the articular cartilage repair tissue. PMID- 15544912 TI - Diagnostic value of diuretic-enhanced excretory MR urography in patients with obstructive uropathy. AB - PURPOSE: Ultrasonography and conventional intravenous urography are most common methods in diagnosis of obstructive uropathies. The disadvantage of ultrasonography is inability of visualizing middle and lower one thirds of ureter, while intravenous urography is using radiation, also functionally extra loading effect on kidneys. In this study, the diagnostic value of MR urography on obstructive uropathy were investigated. MATERIALS AND METHODS: Forty five patients who were suffered from obstructive uropathy examined by ultrasonography, intravenous urography and diuretic-enhanced excretory MR urography by using MR contrast-agent. RESULTS: MR urography established accuracy rate of 92.8% for stone diseases which formed the largest group in this study, however, in other causes of obstructive uropathy, MR urography provide 100% correct diagnosis. CONCLUSION: MR urography provide high quality images for diagnosing and determining causes of urinary obstruction defining position and severity of dilatations as well as showing localization of the pathology. We think that MR urography should be a primary investigation in patients with obstructive uropathy who have contrast agent and X-ray contrindication. PMID- 15544913 TI - Metal-induced oxidative stress and signal transduction. AB - Occupational and environmental exposures to metals are associated with the development of various cancers. Although carcinogenesis caused by metals has been intensively investigated, the mechanisms of action, especially at the molecular level, are still unclear. Accumulating evidence indicates that reactive oxygen species generated by metals may play an important role in the etiology of disease. This review covers recent advances in (1) metal-induced generation of reactive oxygen species; (2) the receptors, kinases, and nuclear transcription factors affected by metals and metal-induced oxidative stress, including growth factor receptors, src kinase, ras signaling, mitogen-activated protein kinases, the phosphoinositide 3-phosphate/Akt pathway, nuclear transcription factor kappaB, activator protein 1, p53, nuclear factor of activated T cells, and hypoxia-inducible factor 1; and (3) global cellular phenomena (signal transduction, cell cycle regulation, and apoptosis) associated with metal-induced ROS production and gene expression. PMID- 15544914 TI - Nitric oxide: a signaling molecule against mitochondrial permeability transition- and pH-dependent cell death after reperfusion. AB - Reperfusion of ischemic tissue can precipitate cell death. Much of this cell killing is related to the return of physiological pH after the tissue acidosis of ischemia. The mitochondrial permeability transition (MPT) is a key mechanism contributing to this pH-dependent reperfusion injury in hepatocytes, myocytes, and other cell types. When ATP depletion occurs after the MPT, necrotic cell death ensues. If ATP levels are maintained, at least in part, the MPT initiates apoptosis caused by mitochondrial swelling and release of cytochrome c and other proapoptotic factors. Cyclosporin A and acidotic pH inhibit opening of permeability transition pores and protect cells against oxidative stress and ischemia/reperfusion injury, whereas Ca(2+), mitochondrial reactive oxygen species, and pH above 7 promote mitochondrial inner membrane permeabilization. Reperfusion with nitric oxide (NO) donors also blocks the MPT via a guanylyl cyclase and protein kinase G-dependent signaling pathway, which in turn prevents reperfusion-induced cell killing. In isolated mitochondria, a combination of cGMP, cytosolic extract, and ATP blocks the Ca(2+)-induced MPT, an effect that is reversed by protein kinase G inhibition. Thus, NO prevents pH-dependent cell killing after ischemia/reperfusion by a guanylyl cyclase/cGMP/protein kinase G signaling cascade that blocks the MPT. PMID- 15544915 TI - Mitochondrial dysfunction and oxidative stress: cause and consequence of epileptic seizures. AB - Mitochondrial dysfunction has been implicated as a contributing factor in diverse acute and chronic neurological disorders. However, its role in the epilepsies has only recently emerged. Animal studies show that epileptic seizures result in free radical production and oxidative damage to cellular proteins, lipids, and DNA. Mitochondria contribute to the majority of seizure-induced free radical production. Seizure-induced mitochondrial superoxide production, consequent inactivation of susceptible iron-sulfur enzymes, e.g., aconitase, and resultant iron-mediated toxicity may mediate seizure-induced neuronal death. Epileptic seizures are a common feature of mitochondrial dysfunction associated with mitochondrial encephalopathies. Recent work suggests that chronic mitochondrial oxidative stress and resultant dysfunction can render the brain more susceptible to epileptic seizures. This review focuses on the emerging role of oxidative stress and mitochondrial dysfunction both as a consequence and as a cause of epileptic seizures. PMID- 15544916 TI - Oxidative lipidomics of apoptosis: redox catalytic interactions of cytochrome c with cardiolipin and phosphatidylserine. AB - The primary life-supporting function of cytochrome c (cyt c) is control of cellular energetic metabolism as a mobile shuttle in the electron transport chain of mitochondria. Recently, cyt c's equally important life-terminating function as a trigger and regulator of apoptosis was identified. This dreadful role is realized through the relocalization of mitochondrial cyt c to the cytoplasm where it interacts with Apaf-1 in forming apoptosomes and mediating caspase-9 activation. Although the presence of heme moiety of cyt c is essential for the latter function, cyt c's redox catalytic features are not required. Lately, two other essential functions of cyt c in apoptosis, that may rely heavily on its redox activity have been suggested. Both functions are directed toward oxidation of two negatively charged phospholipids, cardiolipin (CL) in the mitochondria and phosphatidylserine (PS) in the plasma membrane. In both cases, oxidized phospholipids seem to be essential for the transduction of two distinctive apoptotic signals: one is participation of oxidized CL in the formation of the mitochondrial permeability transition pore that facilitates release of cyt c into the cytosol and the other is the contribution of oxidized PS to the externalization and recognition of PS (and possibly oxidized PS) on the cell surface by specialized receptors of phagocytes. In this review, we present a new concept that cyt c actuates both of these oxidative roles through a uniform mechanism: its specific interactions with each of these phospholipids result in the conversion and activation of cyt c, transforming it from an innocuous electron transporter into a calamitous peroxidase capable of oxidizing the activating phospholipids. We also show that this new concept is compatible with a leading role for reactive oxygen species in the execution of the apoptotic program, with cyt c as the main executioner. PMID- 15544917 TI - The importance of high-density lipoproteins for paraoxonase-1 secretion, stability, and activity. AB - The association of paraoxonase-1 (PON1) with high-density lipoproteins (HDL) is a prerequisite for maintaining normal serum activity of the enzyme. The lipoprotein furnishes an amphipathic environment to shield the hydrophobic, N-terminal region of the enzyme, and such an environment may also be necessary for interaction of PON1 with its substrates. HDL provides the optimal physiological acceptor complex, in terms of both stimulating PON1 secretion and stabilizing the secreted peptide. Lipid and peptide components of HDL contribute to these effects, such that modulating HDL composition influences PON1 activity and function. In this context, understanding how PON1 associates with HDL, what governs the association, and the mechanism by which the PON1-HDL complex exerts its antioxidant function is of particular physiological relevance. Moreover, HDL is subject to substantial compositional variations under both normal and pathological metabolic conditions. It has implications for the influence of the enzyme on cardiovascular risk, as normal enzyme activity may not correlate with optimal functional (antioxidant) efficiency. We review evidence that HDL lipid and protein components interact to promote PON1 secretion and maintain serum enzyme activity. Emerging data on how the enzyme associates with HDL are discussed, and the consequences for PON1 function of modifications to HDL are outlined. Finally, we highlight questions concerning the HDL-PON1 association that remain unanswered but are of particular importance in defining PON1 efficiency. PMID- 15544918 TI - Heme oxygenase expression in human central nervous system disorders. AB - In the normal mammalian CNS, heme oxygenase-2 (HO-2) is constitutively, abundantly, and fairly ubiquitously expressed, whereas heme oxygenase-1 (HO-1) mRNA and protein are confined to small populations of scattered neurons and neuroglia. Unlike ho-2, the ho-1 gene in neural (and many systemic) tissues is exquisitely sensitive to upregulation by a host of pro-oxidant and other noxious stimuli. In Alzheimer disease, HO-1 immunoreactivity is significantly augmented in neurons and astrocytes of the hippocampus and cerebral cortex relative to age matched, nondemented controls and colocalizes to senile plaques, neurofibrillary tangles, and corpora amylacea. In Parkinson disease, HO-1 decorates Lewy bodies of affected dopaminergic neurons and is highly overexpressed in astrocytes residing within the substantia nigra. The ho-1 gene is also upregulated in glial cells within multiple sclerosis plaques; in the vicinity of human cerebral infarcts, hemorrhages, and contusions; and in various other degenerative and nondegenerative human CNS disorders. The products of the heme oxygenase reaction, free ferrous iron, carbon monoxide, and biliverdin/bilirubin, are all biologically active molecules that may profoundly influence tissue redox homeostasis under a wide range of pathophysiological conditions. Evidence adduced from whole animal and in vitro studies indicates that enhanced HO-1 activity may either ameliorate or exacerbate neural injury, effects likely contingent upon the specific model employed, the duration and intensity of HO-1 induction, and the chemistry of the local redox microenvironment. HO-1 hyperactivity also promotes mitochondrial sequestration of nontransferrin iron in oxidatively challenged astroglia and may thereby contribute to the pathological iron deposition and bioenergetic failure amply documented in aging and degenerating human neural tissues. PMID- 15544919 TI - Effect of hydrogen peroxide in redox status estimation using nitroxyl spin probe. AB - A procedure for estimating in vivo redox status using EPR and a hydrogen peroxide (H(2)O(2))-dependent spin probe method is described. The mechanism of decreasing spin clearance in the selenium-deficient (SeD) rat is discussed. The in vivo decay constant of the nitroxyl spin probe in the liver region of SeD rats appeared to be slightly lower that of the selenium-adequate control (SeC) group, and was significantly smaller than that of normal rats. Bile H(2)O(2) levels in normal rats were significantly lower than those in SeD rats. The in vivo decay constant of the spin probe in SeD rats depended on the bile H(2)O(2) level. Furthermore, H(2)O(2) was detected in the bile in all SeD rats, whereas bile H(2)O(2) could be detected in only half of the normal rats. It was found that the in vivo decay constant of the spin probe in normal rats also depended on whether bile H(2)O(2) was detected or not. In vivo decay constants were smaller in rats subjected to the surgical operation than in the nonoperated groups. The EPR signal of the nitroxyl radical in the liver homogenate was increased by addition of H(2)O(2), which was administered 30 min before the rat was killed. It appears that H(2)O(2) can oxidize the hydroxylamine formed following reduction of the spin probe in the liver. PMID- 15544920 TI - Mass spectral evidence for carbonate-anion-radical-induced posttranslational modification of tryptophan to kynurenine in human Cu, Zn superoxide dismutase. AB - Previously, we showed that oxidation of tryptophan-32 (Trp-32) residue was crucial for H(2)O(2)/bicarbonate (HCO(3)(-))-dependent covalent aggregation of human Cu,Zn SOD1 (hSOD1). The carbonate anion radical (CO(3)(-))-induced oxidation of Trp-32 to kynurenine-type oxidation products was proposed to cause the aggregation of hSOD1. Here we used the matrix-assisted laser desorption ionization-time of flight mass spectroscopy, high-performance liquid chromatography-electrospray ionization mass spectroscopy, and liquid chromatography mass spectroscopy methods to characterize products. Results show that a peptide region (31-36) of hSOD1 containing the Trp-32 residue (VWGSIK) is oxidatively modified to the N-formylkynurenine (NFK)- and kynurenine (Kyn) containing peptides (V(NFK)GSIK) and (V(Kyn)GSIK) during HCO(-)-dependent peroxidase activity of hSOD1. Also, UV photolysis of a cobalt complex that generates authentic CO(3)(-) radical induced a similar product profile from hSOD1. Similar products were obtained using a synthetic peptide with the same amino acid sequence (i.e., VWGSIK). We propose a mechanism involving a tryptophanyl radical for CO(3)(-)-induced oxidation of Trp-32 residue (VWGSIK) in hSOD1 to V(NFK)GSIK and V(Kyn)GSIK. PMID- 15544921 TI - Anthraquinones sensitize tumor cells to arsenic cytotoxicity in vitro and in vivo via reactive oxygen species-mediated dual regulation of apoptosis. AB - Cellular oxidation/reduction state affects the cytotoxicity of a number of chemotherapeutic agents, including arsenic trioxide. Reactive oxygen species (ROS), the major intracellular oxidants, may be a determinant of cellular susceptibility to arsenic. Our previous studies showed that a naphthoquinone and an anthraquinone (emodin) displayed the capability of producing ROS and facilitating arsenic cytotoxicity in both leukemia and solid tumor cell lines. We therefore attempted to test emodin and several other kinds of anthraquinone derivatives on EC/CUHK1, a cell line derived from esophageal carcinoma, and on a nude mouse model, with regard to their effects and mechanisms. Results showed that anthraquinones could produce ROS and sensitize tumor cells to arsenic both in vivo and in vitro. The combination of emodin and arsenic promoted the major apoptotic signaling events, i.e., the collapse of the mitochondrial transmembrane potential, the release of cytochrome c, and the activation of caspases 9 and 3. Meanwhile a combination of emodin and arsenic suppressed the activation of transcription factor NF-kappaB and downregulated the expression of a NF-kappaB specific antiapoptotic protein, survivin. These two aspects could be antagonized by the antioxidant N-acetyl-L-cysteine. Therefore anthraquinones exert their effects via a ROS-mediated dual regulation, i.e., the enhancement of proapoptosis and the simultaneous inhibition of antiapoptosis. In vivo study showed that emodin made the EC/CUHK1 cell-derived tumors more sensitive to arsenic trioxide with no additional systemic toxicity and side effects. Taken together, these results suggest an innovative and safe chemotherapeutic strategy that uses natural anthraquinone derivatives as ROS generators to increase the susceptibility of tumor cells to cytotoxic therapeutic agents. PMID- 15544922 TI - Antioxidant effects of water- and lipid-soluble nitroxide radicals in liposomes. AB - Liposomes are today useful tools in different fields of science and technology. A lack of stability due to lipid peroxidation is the main problem in the extension of the use of these formulations. Recent investigative works have reported the protective effects of stable nitroxide radicals against oxidative processes in different media and under different stress conditions. Our group has focused its attention on the natural aging of liposomes and the protection provided by the water- and lipid-soluble nitroxide radicals 2,2,6,6-tetramethylpiperdine-1-oxyl (TEMPO) and doxylstearic acids (5-DSA, 12-DSA, and 16-DSA), respectively. Unilamellar liposomes were incubated under air atmosphere at 37 degrees C, both in the absence and in the presence of these radicals. Conjugated dienes, lipid hydroperoxides, TBARS, membrane fluidity, and nitroxide ESR signal intensity were followed as a function of time. Our results demonstrated that doxylstearic acids were more efficient than TEMPO in retarding lipid peroxidation at all the concentrations tested. The inhibition percentages, depending on the total nitroxide concentration, were not proportional to the lipid-water partition coefficient. Furthermore, time-course ESR signals showed a slower decrease for doxylstearic acids than for TEMPO. No significant differences were found among 5 DSA, 12-DSA, and 16-DSA. We concluded that the nitroxide radical efficiency as antioxidant directly depends on both nitroxide concentration and lipophilicity. PMID- 15544923 TI - Paraoxonase 2 (PON2) expression is upregulated via a reduced-nicotinamide-adenine dinucleotide-phosphate (NADPH)-oxidase-dependent mechanism during monocytes differentiation into macrophages. AB - Paraoxonase 2 (PON2) is a member of the paraoxonases gene family. PON2 is ubiquitously present in cells, including macrophages, and it was shown to protect against cellular oxidative stress. The aim of the present study was to analyze mechanisms involved in PON2 expression during monocyte/macrophage differentiation. PON2 expression was analyzed in vitro in THP-1 cells differentiated with 1alpha,25-dihydroxyvitamin D3 and in vivo in mouse peritoneal macrophages (MPM) isolated at increasing time intervals after intraperitoneal thioglycollate injection. PON2 expression (mRNA and protein) and activity gradually increased during monocyte/macrophage differentiation, up to five fold and eight fold in vitro and in vivo, respectively. This effect was associated with a gradual increase in cellular superoxide anion production. Supplementation of vitamin E to Balb/C mice inhibited the reduced nicotinamide adenine dinuleotide phosphate (NADPH)-oxidase-dependent increase in cellular superoxide anion production by 50% and down-regulated PON2 mRNA expression and activity by 30 and 60%, respectively. Furthermore, PON2 expression was lower by nine fold in MPM isolated from P47(phox-/-) (inactive NADPH oxidase) mice, in comparison to MPM from control mice. PON2 expression was found to be regulated, at least in part, by the transcription factor AP-1, as suggested by decreased JDP2 (AP-1 repressor) protein expression in the nucleus and by decreased PON2 expression in the presence of a Jun N-terminal kinase inhibitor (SP600125). The present study demonstrates, for the first time, that PON2 expression increases in monocytes during their maturation into macrophage as a result of NADPH-oxidase activation, and this process is partly regulated by the transcription factor AP-1. PON2 stimulation may represent a compensatory mechanism against the increase in cellular superoxide anion production and atherogenesis. PMID- 15544924 TI - The antioxidant defense protein heme oxygenase 1 is a novel target for statins in endothelial cells. AB - Cholesterol-independent, pleiotropic actions of hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) exert anti-inflammatory and antioxidant action by as yet unidentified mechanisms. This study explores the role of heme oxygenase 1 (HO-1) as a target and mediator of statins. In cultured endothelial cells derived from human umbilical vein, simvastatin and lovastatin increased HO 1 mRNA levels in a concentration- and time-dependent fashion. HO-1 induction by statins remained unaffected by mevalonate and N-nitro-l-arginine methyl ester, precluding the involvement of isoprenoid- and NO-dependent pathways. HO-1 mRNA induction was abrogated in the presence of actinomycin D and cycloheximide. In cells transfected with a reporter gene construct containing the proximal 4 kB of the HO-1 gene promoter 5'-flanking region, significant upregulation of promoter activity was detected, indicating that regulatory elements binding to this region were involved in transcriptional HO-1 induction by statins. Increased transcriptional expression of HO-1 was associated with elevated HO-1 protein levels and reduction of free radical formation. Our results show that the antioxidant defense protein HO-1 is a target site of statins in endothelial cells. Statins lead to HO-1 promoter activation, transcript and protein accumulation. This novel pathway may contribute to and explain the pleiotropic antioxidant, anti-inflammatory, and antiatherogenic actions of statins. PMID- 15544925 TI - Oxidative modification of mitochondrial respiratory complexes in response to the stress of Trypanosoma cruzi infection. AB - Previously, we have shown deficiencies in the activities of the mitochondrial respiratory complexes and reduced mitochondrial ATP generation capacity in chagasic hearts infected by Trypanosoma cruzi. In this study, we determined whether the oxidative stress that occurs in response to T. cruzi infection contributes to the catalytic impairment of respiratory complexes and to subsequent mitochondrial dysfunction in murine myocardium. Our data show that oxidative injuries, as determined by the levels of lipid peroxides and protein carbonyls, are incurred in cardiac mitochondria as early as 3 days postinfection and persist throughout the infection and disease. The individual components of the respiratory complexes were separated by two-dimensional, blue-native gel electrophoresis, and carbonyl adducts were detected by Western blotting. We observed substantial carbonylation of the specific subunits of mitochondrial respiratory complexes in infected murine hearts. Of note is the oxidative modification of NDUFS1, NDUFS2, and NDUFV1, which form the catalytic core of the CI complex; UQCRC1, UQCRC2, and UQCRQ, the subunits of the core subcomplex, and UQCRH and CYC1, which form the cyt c(1) subcomplex of CIII; and a gamma chain that is essential for ATP synthesis by CV complex. The extent of oxidative modifications of the subunits correlated with the catalytic defects of the respiratory complexes in the infected myocardium. Taken together, our data demonstrate that respiratory complexes are oxidatively damaged in response to the stress of T. cruzi infection. These data also suggest involvement of the specific susceptibility of the protein subunits, and not generalized mitochondrial oxidative damage in respiratory chain impairment of chagasic hearts. PMID- 15544926 TI - Drug delivery systems for oestrogenic hormones and antagonists: the need for selective targeting in estradiol-dependent cancers. AB - The pleiotropic activity of oestrogens and their mechanism of action via their binding to the two oestrogen receptors alpha (ER alpha) and beta (ER beta) subtypes in the different tissues where oestrogens exert their action have been briefly described. The fate of these compounds trapped into different galenic forms is discussed with regard to their therapeutic applications. Firstly, the advantages and disadvantages of the different forms (pills, i.v. forms and transdermal patches) used in contraception are compared. Secondly, the therapeutic use of formulated oestrogens for the post-menopausal hormone replacement therapy (HRT) is analysed through the various results obtained in different trials. The link between HRT and the risks of breast cancer and cardiovascular disease is underlined. Finally, comparing the activity of selective oestrogen receptor modulators such as tamoxifen and pure anti oestrogens such as RU58668 and ICI182780, we analysed the reasons leading to the need for a tumor targeting of the latters, but not of the former for the treatment of oestrogen-dependent breast cancer. Different injectable and biodegradable formulations, that lead to a remarkable anti-tumor efficiency in xenografts, have been recently developed and we believe that they may represent promising new administration ways of added therapeutic values for anti oestrogens. Such devices could be extended to the delivery of other anti-cancer drugs with more aggressive activities than anti-oestrogens. PMID- 15544927 TI - Molecular cloning and characterization of two novel human RXR alpha splice variants. AB - Two novel cDNAs encoding RXR alpha splice variants (RXR alpha 2 and RXR alpha 3) were identified among human full-length cDNA libraries. RXR alpha 2 and RXR alpha 3 cDNAs possess open reading frames, leading to production of proteins lacking the N-terminal 27 and 97 amino acid residues of the RXR alpha 1 product, respectively. RXR alpha 2 and RXR alpha 3 genes have respective 5'-terminal exons. RXR alpha 3 is expressed in brain, spleen and prostate whereas the expression of RXR alpha 2 was below the detectable level. Both RXR alpha 2 and RXR alpha 3 showed a level of transcriptional activity and a dose response curve against the agonist LG100268 similar to RXR alpha 1 in reporter assay for the RXR alpha homodimer or that for the heterodimer with PPAR gamma 2. However, clear differences were observed among the splice variants when dose response curves were compared by the assay in the presence of coactivators such as SRC-1 and PGC 1. These results suggest specific physiological roles of two novel human RXR alpha splice variants. PMID- 15544928 TI - 6-[2-(adamantylidene)-hydroxybenzoxazole]-O-sulfamate, a steroid sulfatase inhibitor for the treatment of androgen- and estrogen-dependent diseases. AB - Steroid sulfatase (STS) offers a new target for the treatment of steroid hormone dependent diseases, such as breast and prostate cancer and androgen-dependent skin diseases. We here characterize a novel non-estrogenic inhibitor of the enzyme, namely 6-[2-(adamantylidene)-hydroxybenzoxazole]-O-sulfamate (AHBS), with special attention to its potential use in the treatment of acne. The compound blocks STS activity in homogenates of human skin with IC(50)=16 nM. Following a single oral dose (5 mg/kg) in rats, the compound blocks STS in the skin by 95% at 8 h, followed by recovery of activity over 5 days. Following topical application to the skin, both in vitro and in vivo, AHBS passes through the stratum corneum leading to inhibition of STS activity in the dermal compartment with rapid onset and long duration. Topical application of AHBS to Gottingen minipigs for a period of 2 weeks does not induce symptoms of ichthyosis as seen in STS-deficient human subjects, but leads to a reduction of sebum secretion to the skin surface. Based on these data, clinical studies with AHBS in acne patients are warranted, in order to verify the hypothesis on the importance of the sulfatase pathway in androgen-dependent skin diseases. PMID- 15544929 TI - Two distinct cytochrome P450 aromatases in the orange-spotted grouper (Epinephelus coioides): cDNA cloning and differential mRNA expression. AB - The cDNA sequences encoding two distinct cytochrome P450 aromatases, namely P450aromB and P450aromA, were isolated from brain and ovary cDNA libraries of the orange-spotted grouper, respectively. The P450aromB cDNA consists of 1892 bp, and the open reading frame (ORF) encodes a putative protein of 506 amino acids. The P450aromA cDNA consists of 1836 bp, and the ORF encodes a putative protein of 518 amino acids. Northern blot analysis revealed a transcript of about 1.9 kb for P450aromB in the brain and kidney, and 2.1 kb for P450aromA in the ovary. The expression of both P450aromB and P450aromA genes in different tissues was further examined using one-step RT-PCR followed by Southern blot analysis. High levels of P450aromB mRNA expression were detected in the olfactory bulb, forebrain, midbrain, hypothalamus, medulla, pituitary, gill filament, gill arch, kidney, muscle, adipose tissue, and blood cells, but low levels in the hindbrain and ovary. High levels of P450aromA mRNA expression were detected in the ovary, pituitary, gill filament, gill arch, and spleen, but low levels in the forebrain, hindbrain, hypothalamus, and blood cells. In addition, the expression of P450arom genes in the orange-spotted grouper of different gonadal stages as induced by 17 alpha-methyltestosterone (MT) was investigated. The mRNA expression of P450aromB in the hypothalamus was highest in the intersexual stage, whereas the mRNA expression of P450aromA in the gonads was highest in the female stage, decreased in the intersexual stage, and lowest in the male stage. Results from current study indicate that P450aromB and P450aromA genes of the orange-spotted grouper have distinct tissue patterns of mRNA expression, and both of them may be involved in the MT-induced sex change. PMID- 15544930 TI - Non-classical regulation of estrogen receptor-alpha by ICI182,780. AB - Estrogen receptor-alpha (ER alpha) regulates transcription through a number of molecular mechanisms. Two mechanisms by which ER alpha acts directly in the nucleus have emerged: (1) in classical ER alpha action, estrogen-bound receptor binds estrogen response elements (ERE) and regulates promoters by recruiting coactivators or corepressors to DNA; (2) non-classical action is not dependent on ER alpha binding to EREs; its mechanism is not as clearly defined as classical action. In many instances, non-classical action is mediated by tethering of ER alpha to other DNA-binding proteins, facilitating recruitment of coregulators to transcription regulatory sequences. In some cell types, non-classical stimulation can be enhanced by antagonists and repressed by agonists of ER alpha. Here, we show that non-classical action of ER alpha in 293 cells occurs in a wide range of enhancers and enhancer binding proteins. ER alpha stimulates AP-1 elements, cyclic AMP response elements (CRE), and serum response elements (SRE) in the presence of the antiestrogen ICI182,780. Further, in the presence of ICI182,780, ER alpha stimulates activation domains of Jun, ATF-2, Elk, and CRE-binding protein (CREB). Non-classical ER alpha regulation described here does not appear to be sensitive to point mutations which affect classical and tethered ER alpha action; moreover, in our experiments, non-classical action is uniquely sensitive to nuclear transport inhibition by leptomycin B. Because ICI182,780 appears to affect multiple and diverse transcriptional systems, our results are likely explained by ER alpha-dependent modulation of common components of the transcriptional machinery and may not be completely explained by tethering of ER alpha to specific transcription factors. PMID- 15544931 TI - Cyclin D1 expression is dependent on estrogen receptor function in tamoxifen resistant breast cancer cells. AB - The development of resistance to tamoxifen, the most common antiestrogen used in the treatment of breast cancer, is a frequent and severe clinical problem. Tamoxifen-resistant tumors are still capable of responding to other hormonal therapies such as those that downregulate estrogen receptor expression. Mechanisms leading to acquisition of tamoxifen-resistant but hormone-sensitive growth are not completely understood. In tamoxifen-sensitive breast cancer cells, tamoxifen inhibits, whereas estrogen induces, expression of cyclin D1, a key cell cycle regulatory protein. Ectopic expression of cyclin D1 can lead to antiestrogen resistance. Thus, to determine whether cyclin D1 is involved in the growth of tamoxifen-resistant cells, we developed several tamoxifen-resistant variants from MCF-7 cells. These variants grow in the absence of estrogen or in the presence of tamoxifen, but their growth is inhibited by estrogen receptor downregulators. We show here that cyclin D1 expression is maintained at comparable levels in all tamoxifen-resistant variants, whereas pS2, another estrogen-regulated protein, is not. The addition of physiological levels of estrogen further stimulates cyclin D1 expression and proliferation. In contrast, treatment with estrogen receptor downregulators decreases cyclin D1 expression and proliferation. Thus, changes in cyclin D1 expression upon second-line hormonal therapy may predict hormonal sensitivity of tamoxifen-resistant tumors. These studies suggest that estrogen receptor mediates cyclin D1 expression and growth of tamoxifen-resistant tumors. PMID- 15544932 TI - Detection of thiopental in the steroid fraction of serum from neonates following maternal exposure. AB - To follow up an investigation which studied effects of antenatal dexamethasone therapy on neonatal respiratory performance in multifetal gestations, neonatal serum steroids were determined by HPLC. A major peak (X) whose retention time coincided with that of dexamethasone was observed in many, but not all, serum samples. However, there was no correlation between the neonates whose serum samples displayed this X-peak and the mothers who had actually received the steroid therapy, indicating that the X-substance was not dexamethasone. An alternate mobile phase was employed which separated the X-substance and dexamethasone validating the indication. Among ten clinical conditions of the neonate birth, the X-substance was found to correlate only with the mothers who had the cesarean operation for delivery, suggesting that the substance was not necessarily a steroid. Four anesthetic agents used for cesarean operations were studied; the X-substance was identified as thiopental using a LC/MS technique. This was based on the same retention times, the same negative ions at m/z 240.9 and the same daughter ions at m/z 100.8 between the two substances. Thus, caution must be exercised when HPLC is employed to study serum steroids of patients who have previously been exposed to thiopental. Moreover, recent reports have shown that thiopental affects certain metabolic reactions in the rat; the present findings also suggest a need for further investigations of thiopental effect on neonates. PMID- 15544934 TI - Ras activation is associated with vitamin D receptor mRNA instability in HC11 mammary cells. AB - HC11, a spontaneously immortalized murine mammary lineage maintains features of normal cells while HC11 H-ras transformed cells (HC11 ras) are tumorigenic. Ras transformation is associated with a lower Vitamin D receptor (VDR) mRNA content. Our goal was to investigate the mechanism underlying VDR mRNA differences between these cells. Although the VDR transcriptional rate measured by run-on assays did not differ between the cells, our data suggested a pos transcriptional mechanism involving higher VDR mRNA degradation in HC11 ras cells which was not due to mutations in its 3'-UTR region since sequences of mRNA obtained from HC11 and HC11 ras cells were identical. Treatment of HC11 ras cells with a farnesyltransferase inhibitor, which prevents ras activation, causing an enhancement of VDR mRNA levels, indicating an association between the ras signaling pathway and VDR mRNA instability. The present work suggests that the decreased mRNA levels in HC11 ras cells might in part be due to an early loss of stability. PMID- 15544933 TI - The selective estrogen receptor modulator SCH 57068 prevents bone loss, reduces serum cholesterol and blocks estrogen-induced uterine hypertrophy in ovariectomized rats. AB - Our objective was to determine the effects of SCH 57068 alone and with 17 beta estradiol (E(2)) on bone, lipids and uteri in ovariectomized (OVX) rats. In OVX animals lumbar vertebral and femoral bone mineral density (BMD) were significantly higher after 12 weeks of treatment with SCH 57068 than in untreated OVX controls. Similarly BMD was superior in OVX + E(2) + SCH 57068 treated animals than in OVX + E(2) controls. SCH 57068 also significantly reduced the increase in bone turnover markers, serum pyridinoline and serum osteocalcin levels, induced by OVX, and increased mechanical bone strength. SCH 57068 also significantly reduced the rise in serum cholesterol and low-density lipoprotein cholesterol induced by OVX. SCH 57068 had no stimulatory effect on uterine epithelium when given alone in OVX rats. SCH 57068 (1 and 2.5 mg/kg) reduced uterine weight and blocked endometrial stimulation induced by E(2). In summary, SCH 57068 adds to the positive effects of E(2) on bone and lipid metabolism but blocks the stimulatory effects of E(2) on the uterus. Potentially, E(2) + SCH 57068 could be combined for the treatment and prevention of breast cancer or as a novel hormone replacement therapy. PMID- 15544935 TI - Differential distribution of splice variants of estrogen receptor beta in human testicular cells suggests specific functions in spermatogenesis. AB - A growing number of estrogen receptor beta (ER beta) splice variants are reported. Several of these have been discovered in testis, but with few exceptions little is known about their cellular localization. The aim of this study was to identify and elucidate the mRNA expression pattern of the different ER beta splice variants in human testicular cells. Northern analysis was performed on whole testis and fractions enriched in germ cells from untreated men and from estrogen-treated men undergoing sex change surgery. Probes were constructed in order to systematically screen for and identify various ER beta splice variants. Several ER beta bands were observed in the human testis, in which splice variants constituted the major part of total ER beta transcripts. Interestingly, only two ER beta wild-type transcripts were detected. These seem to be virtually absent from the haploid germ cells and are probably mainly located in somatic cells and/or primary spermatocytes. Several novel ER beta deletion variants were found in high levels in the haploid germ cell fractions and were nearly absent in testicular cells from the estrogen-treated men. The cell-dependent distribution raises the question whether splice variants may have specific functions in spermatogenesis, and whether the differential splicing of ER beta is regulated in a cell-specific manner. PMID- 15544936 TI - Two major metabolites of 8-prenylnaringenin are estrogenic in vitro. AB - 8-prenylnaringenin (8-PN) and preparations containing 8-prenylnaringenin have been suggested for use in medicinal and cosmetic applications like hormone replacement or bust enhancement. However, the safety of application is still under considerable debate. Recently it has been shown that human liver microsomes are converting 8-prenylnaringenin to 12 metabolites, with (E)-8-(4'' hydroxyisopentenyl)naringenin (8-PN-OH) and (E)-8-(4''-oxoisopentenyl)naringenin (8-PN=O) being among the most abundant. Applying two independent in vitro test systems we demonstrate that these two metabolites of 8-prenylnaringenin are estrogenic in vitro. These results represent an important piece of information towards the discussion of safety of use of preparations containing 8 prenylnaringenin. PMID- 15544937 TI - Immune-responsive lysozymes from hemocytes of the American dog tick, Dermacentor variabilis and an embryonic cell line of the Rocky Mountain wood tick, D. andersoni. AB - Immune-responsive lysozyme encoding cDNAs were identified from two medically important tick species by an expressed sequence tag approach of D. variabilis hemocytes (Dv Lys) and a D. andersoni embryonic derived cell line, DAE100. Comparative sequence analyses indicated the Dermacentor molecules to be products of orthologous genes and to be most similar to arthropod c-type lysozymes. Northern blotting analyses demonstrated that Dv Lys expression levels were most abundant in tick hemocytes and to a much lesser degree in the midgut while barely detectable in ovary, salivary gland, and Malpighian tubule tissues. Involvement of the Dermacentor c-type lysozymes in innate immunity was demonstrated by Escherichia coli challenges of D. variabilis ticks by injection resulting in a temporal profile of significantly elevated transcript abundances above those of naive controls that was similarly observed of the D. andersoni cells co-cultured with E. coli. In contrast to that reported of the digestive gut lysozyme of the soft tick Ornithodoros moubata, Dv Lys levels were not statistically differentially regulated by blood meal digestion. Additionally, given the differences in tissue distribution, sequence characteristics and phylogenetic placements between the Dermacentor and Ornithodoros lysozymes demonstrates that ticks possess differently adapted c-type lysozymes that are spatially and temporally differentially expressed. PMID- 15544938 TI - Identification of amine receptors from a swallowtail butterfly, Papilio xuthus L.: cloning and mRNA localization in foreleg chemosensory organ for recognition of host plants. AB - The swallowtail butterfly, Papilio xuthus L., feeds exclusively on members of the plant family, Rutaceae. Female butterflies lay eggs in response to specific chemicals contained in their host plants. They perceive a variety of polar compounds as oviposition stimulants through the tarsal chemosensilla of the foreleg by drumming upon the leaf surface. Some biogenic amine analogs have been characterized as oviposition stimulants. We have cloned three amine receptors, serotonin, tyramine, and dopamine, from cDNA derived from foreleg tarsus of P. xuthus, and determined structures of both cDNA and genomic genes. The phenylethylamine (tyramine and dopamine) receptors were expressed preferentially in brain and chemosensory organs. Moreover, we observed the localized expression of dopamine receptors at the base of tarsal chemosensilla by in situ hybridization. These results suggest that amine receptors in tarsal chemosensilla have a functional role in chemoreception for host plant recognition. PMID- 15544939 TI - Phenoloxidase activity in Apis mellifera honey bee pupae, and ecdysteroid dependent expression of the prophenoloxidase mRNA. AB - Phenoloxidase (monophenol, l-dopa: oxygen oxidoreductase, EC 1.14.18.1) is a multicopper oxidase, which plays an important role in melanin synthesis, necessary for defense against intruding microorganisms and parasites, wound healing and cuticle pigmentation. A phenoloxidase from the hemolymph of honey bee pupae exhibited an apparent molecular mass of 70 kDa, as estimated by gel filtration and SDS-PAGE. Optimal pH and temperature were 6.5 and 20 degrees C, respectively. Activity was fully stable for 30 min at 50 degrees C. Like phenoloxidases from the hemolymph of other insects, the honey bee enzyme was activated by trypsin and inhibited by protease inhibitors and phenylthiourea. Only high concentrations of sodium azide effectively inhibited the detected activity. A low concentration (5 microM) of Ca2+, Mg2+, and Mn2+ had a stimulatory effect on the activity. Single Michaelis-Menten curves were observed for l-dopa and dopamine oxidation, but the affinity of the enzyme for dopamine was greater than for L-dopa. Semiquantitative RT-PCR and Southern blot analysis using a 359 bp labeled probe, and quantification of the prophenoloxidase mRNA levels by real-time PCR showed increased amounts of transcripts in hemocytes and integument from young pupae injected with 20-hydroxyecdysone. PMID- 15544940 TI - cAMP-dependent protein kinase of Manduca sexta phosphorylates but does not activate the fat body triglyceride lipase. AB - cAMP-dependent-protein kinase (PKA) is a central player of the adipokinetic signal that controls the mobilization of stored lipids in the fat body. Previous studies showed that adipokinetic hormone (AKH) rapidly activates PKA from the fat body of Manduca sexta (Arrese et al. (J. Lipid. Res. 40(3): 556)). As a part of our investigation on lipolysis in insects, here we report the purification and characterization of the catalytic subunit of PKA from the fat body of M. sexta and its role in the direct activation of the TG lipase in vitro. PKA was purified to apparent homogeneity and the identity of the protein was confirmed by MALDI TOF and Western blot analysis. The enzyme showed a high affinity for Mg-ATP (Km = 39 microM) and Kemptide (Km = 31 microM) and was strongly inhibited by the PKA specific inhibitors PKI 5-24 and H89. Manduca sexta PKA only recognized serine residues as phosphate acceptor; theronine or tyrosine containing peptides were not phosphorylated. Purified fat body TG-lipase proved to be a good substrate of the purified kinase. However, phosphorylation of the lipase did not enhance the lipolytic activity of the enzyme in vitro. These results suggest that, besides lipase phosphorylation, the mechanism of AKH-induced activation of the lipolysis requires the involvement of other proteins and/or signals. PMID- 15544941 TI - Cloning and expression of the yolk protein of the tsetse fly Glossina morsitans morsitans. AB - Two major families of nutritional proteins exist in insects, namely the vitellogenins and the yolk proteins. While in other insects only vitellogenins are found, cyclorraphan flies only contain yolk proteins. Possible sites of yolk protein synthesis are the fat body and the follicle cells surrounding the oocyte. We report the cloning of the yolk protein of the tsetse fly Glossina morsitans morsitans, a species with adenotrophic viviparity. The tsetse fly yolk protein could be aligned with other dipteran yolk proteins and with some vertebrate lipases. In contrast to the situation in most fly species, only a single yolk protein gene was found in the tsetse fly. Northern blot analysis showed that only the ovarian follicle cells, and not the fat body represents the site of yolk protein synthesis. PMID- 15544942 TI - Processing of pro-thrombostasin by a recombinant subtilisin-like proprotein convertase derived from the salivary glands of horn flies (Haematobia irritans). AB - Thrombostasin (TS) is a thrombin inhibitor found in the salivary glands of horn flies (Haematobia irritans). It is produced as an inactive form with a 76-amino acid propeptide in the N-terminus preceding the mature TS. A minimal recognition sequence by subtilisin-like proprotein convertases, Arg-Xaa-Xaa-Arg, is localized C-terminal to the propeptide. This study demonstrated that a gene cloned from the salivary glands of the horn fly encodes a new convertase, subsequently named horn fly proprotein convertase (HFPC), and that the recombinant HFPC expressed in insect HighFive cell culture specifically cleaves recombinant pro-thrombostasin, produced in E. coli, at the expected site. The relative cleavage efficiency of rHFPC was compared with that of recombinant human furin, a commercially available proprotein convertase. The result indicated that this newly identified proprotein convertase is of importance for the proteolytic maturation of thrombostasin, a protein secreted in horn fly saliva and used by the insect to counteract its host's haemostatic response. PMID- 15544943 TI - A Drosophila salivary gland mucin is also expressed in immune tissues: evidence for a function in coagulation and the entrapment of bacteria. AB - Our studies on the developmental regulation of glycosylation in Drosophila melanogaster led us to identify and characterize gp150, an ecdysone-regulated mucin that is found in hemocytes, the gut (peritrophic membrane) and in the salivary glands. We are particularly interested in mucin immune functions and found that gp150 is released from larval hemocytes, becomes part of the clot and participates in the entrapment of bacteria. By RT-PCR and RNAi experiments, we identified gp150 as the previously described I71-7, an ecdysone-induced salivary glue protein. We discuss the evolutionary and biochemical implications of the dual use of salivary proteins for immune functions in insects. Further molecular characterization of such shared proteins may enable a better understanding of the properties of proteins involved in containment and elimination of microbes, as well as hemostasis and wound repair. PMID- 15544944 TI - Identification of mutations associated with pyrethroid resistance in the para type sodium channel of the cat flea, Ctenocephalides felis. AB - Knockdown resistance (kdr) to pyrethroid insecticides is caused by point mutations in the pyrethroid target site, the para-type sodium channel of nerve membranes. This most commonly involves alterations within the domain II (S4-S6) region of the channel protein where five different mutation sites have been identified across a range of insect species. To investigate the incidence of this mechanism in cat fleas, we have cloned and sequenced the IIS4-IIS6 region of the para sodium channel gene from seven laboratory flea strains. Analysis of these sequences revealed two amino acid replacements at residues previously implicated in pyrethroid resistance. One is the 'common' kdr mutation, a leucine to phenylalanine substitution (equivalent to L1014F of housefly) reported previously in several other insects. The other is a threonine to valine substitution (equivalent to T929V) and is a novel variant of the T929I mutation first identified in diamondback moth. The L1014F mutation was found at varying frequency in all of the laboratory flea strains, whereas the T929V mutation was found only in the highly resistant Cottontail strain. We have developed rapid PCR based diagnostic assays for the detection of these mutations in individual cat fleas and used them to show that both L1014F and T929V are common in UK and US flea populations. This survey revealed a significant number of fleas that carry only the V929 allele indicating that co-expression with the F1014 allele is not necessary for flea viability. PMID- 15544945 TI - Expression and evolution of delta9 and delta11 desaturase genes in the moth Spodoptera littoralis. AB - Desaturation of fatty acids is a key reaction in the biosynthesis of moth sex pheromones. The main component of Spodoptera littoralis sex pheromone blend is produced by the action of Delta11 and Delta9 desaturases. In this article, we report on the cloning of four desaturase-like genes in this species: one from the fat body (Sls-FL1) and three (Sls-FL2, Sls-FL3 and Sls-FL4) from the pheromone gland. By means of a computational/phylogenetic method, as well as functional assays, the desaturase gene products have been characterized. The fat body gene expressed a Delta9 desaturase that produced (Z)-9-hexadecenoic and (Z)-9 octadecenoic acids in a (1:4.5) ratio, whereas the pheromone gland Sls-FL2 expressed a Delta9 desaturase that produced (Z)-9-hexadecenoic and (Z)-9 octadecenoic acids in a (1.5:1) ratio. Although both Delta9 desaturases produced (Z)-9-tetradecenoic acid from myristic acid, transformed yeast grown in the presence of a mixture of myristic and (E)-11-tetradecenoic acids produced (Z,E) 9,11-tetradecadienoic acid, but not (Z)-9-tetradecenoic acid. The Sls-FL3 gene expressed a protein that produced a mixture of (E)-11-tetradecenoic, (Z)-11 tetradecenoic, (Z)-11-hexadecenoic and (Z)-11-octadecenoic acids in a 5:4:60:31 ratio. Despite having all the characteristics of a desaturase gene, no function could be found for Sls-FL4. PMID- 15544946 TI - The role of phenylalanine hydroxylase in melanotic encapsulation of filarial worms in two species of mosquitoes. AB - Melanin formation has a significant influence on mosquito vector competence by limiting the development of metazoan parasites. Tyrosine, the rate-limiting substrate of melanin production, can be obtained exogenously or derived from phenylalanine by phenylalanine hydroxylase (PAH). The characteristics of this defense mechanism, such as temporal expression of constituent enzymes involved in the biosynthetic pathway, can vary considerably between mosquito species. We investigated the functional role of PAH in the melanotic encapsulation response in Aedes aegypti and Armigeres subalbatus, two mosquito species with markedly different melanization responses. We used double-stranded RNA (dsRNA) to knock down PAH and observed the phenotypic effects on melanin formation. PAH transcripts were dramatically reduced in both mosquito species after gene knock down. The abundance of PAH proteins was decreased in gene knockdown mosquitoes that were inoculated with Dirofilaria immitis microfilariae (mf) as compared to inoculation controls. A significant reduction of mf melanization also was observed in these knockdown mosquitoes as compared to inoculation controls. Our data suggest that PAH is required for a fully functional melanotic encapsulation response in both mosquito vectors. PMID- 15544947 TI - Patterns and clusters within the PSM column in TiBS, 1992-2004. AB - Sequence similarities among proteins can infer biological function and evolutionary relationships--a powerful approach for investigating new proteins and suggesting future experiments. The availability of public sequence databases and freely distributed tools for sequence analysis has meant that researchers from all over the world can use this approach. For the past 12 years, the Protein Sequence Motif column in TiBS has provided a platform for documenting interesting discoveries from sequence analyses. As the column comes to an end, we look at the published contributions over the years and reflect on sequence analysis through the beginning of the genomic era. PMID- 15544948 TI - The BACK domain in BTB-kelch proteins. AB - A novel conserved motif--the BACK (for BTB and C-terminal Kelch) domain--is found in the majority of proteins that contain both the BTB domain and kelch repeats. Many kelch-repeat proteins are involved in organization of the cytoskeleton via interaction with actin and intermediate filaments, whereas BTB domains have multiple cellular roles, including recruitment to E3 ubiquitin ligase complexes. The identification of the BACK domain in BTB and kelch proteins, and its high conservation across metazoan genomes, suggest an important function for this domain with a possible role in substrate orientation in Cullin3-based E3 ligase complexes. PMID- 15544949 TI - Rad23 and Rpn10: perennial wallflowers join the melee. AB - Substrate ubiquitination is highly regulated; by contrast, substrate targeting to the proteasome has been considered a stochastic process that is mediated primarily by high-affinity interaction with multi-ubiquitin chains. However, recent findings have shown that substrate recognition by the proteasome is also regulated, and requires Rad23, Dsk2 and Rpn10. These studies suggest that the engagement of protein ubiquitination and translocation with degradation by the proteasome is coordinated by a series of regulated events. PMID- 15544950 TI - Hierarchical thinking in network biology: the unbiased modularization of biochemical networks. AB - As reconstructed biochemical reaction networks continue to grow in size and scope, there is a growing need to describe the functional modules within them. Such modules facilitate the study of biological processes by deconstructing complex biological networks into conceptually simple entities. The definition of network modules is often based on intuitive reasoning. As an alternative, methods are being developed for defining biochemical network modules in an unbiased fashion. These unbiased network modules are mathematically derived from the structure of the whole network under consideration. PMID- 15544951 TI - Quality control of integral membrane proteins. AB - Integral membrane proteins (IMPs) are essential components of the plasma and organellar membranes of the eukaryotic cell. Non-native IMPs, which can arise as a result of mutations, errors during biosynthesis or cellular stress, can disrupt these membranes and potentially lead to cell death. To protect against this outcome, the cell possesses quality control (QC) systems that detect and dispose of non-native IMPs from cellular membranes. Recent studies suggest that recognition of non-native IMPs by the QC machinery is correlated with the thermodynamic stability of these proteins. Consistent with this, small molecules known as chemical and pharmacological chaperones have been identified that stabilize non-native IMPs and enable them to evade QC. These findings have far reaching implications for treating human diseases caused by defective IMPs. PMID- 15544952 TI - Chemoenzymatic synthesis of oligosaccharides and glycoproteins. AB - Oligosaccharides are involved in a wide range of biological processes including, for example, bacterial and viral infection, cancer metastasis, the blood-clotting cascade and many other crucial intercellular recognition events. The molecular details of these biological recognition events are, however, not well understood. To express their function, oligosaccharides often occur as glycoconjugates attached to proteins (called glycoproteins) or lipids (called glycolipids) that are often found on the surface of cells. Such physiological relevance has stimulated researchers to make significant advances in oligosaccharide and glycoprotein preparation despite the chemically imposing and polydisperse nature of these molecules. The chemical and Chemoenzymatic methods developed recently have facilitated the synthesis of structurally defined oligosaccharides and glycoconjugates such that a more thorough understanding of their biological function and potential therapeutic application can be addressed. PMID- 15544953 TI - Enzymes involved in the activation and inactivation of vitamin D. AB - Six cytochrome P450 (CYP) isoforms have been shown to hydroxylate vitamin D. Four of these, CYP27A1, CYP2R1, CYP3A4 and CYP2J3, are candidates for the enzyme vitamin D 25-hydroxylase that is involved in the first step of activation. The highly regulated, renal enzyme 25-hydroxyvitamin D-1alpha-hydroxylase contains the component CYP27B1, which completes the activation pathway to the hormonal form 1alpha,25-dihydroxyvitamin D(3). A five-step inactivation pathway from 1alpha,25-(OH)(2)D(3) to calcitroic acid is attributed to a single multifunctional CYP, CYP24A1, which is transcriptionally induced in vitamin D target cells by the action of 1alpha,25-(OH)(2)D(3). On the basis of alignments and crystal structures of other CYPs, homology models of vitamin-D-related CYPs have been generated. Two human forms of rickets caused by mutations of CYP2R1 and CYP27B1, as well as mouse knockout models of CYP27A1, CYP27B1 and CYP24A1, are helping us to establish the full in vivo physiological roles of the vitamin-D related hydroxylases. PMID- 15544954 TI - Rpb4 and Rpb7: subunits of RNA polymerase II and beyond. AB - RNA Polymerase II (pol II) is a large multi-subunit complex that is responsible for the synthesis of all eukaryotic mRNAs. Its correct and timely recruitment to promoter regions is a crucial step of transcription regulation, involving complicated and well-controlled networks of protein-DNA and protein-protein interactions. The best-studied pol II is the yeast complex consisting of 12 subunits (Rpb1-12). Rpb4 and Rpb7 form a dissociable heterodimer (Rpb4/7). The unique location of Rpb4/7 within the transcription initiation complex, and its capacity to interact with various transcription factors, suggest that it provides important links to the network of interactions that control transcription initiation. Moreover, Rpb4/7 executes some non-transcriptional activities, including mRNA transport. Hence, Rpb4/7 functions at the interface of transcriptional and post-transcriptional machinery. PMID- 15544955 TI - Longins and their longin domains: regulated SNAREs and multifunctional SNARE regulators. AB - Longins are the only R-SNAREs that are common to all eukaryotes and are characterized by a conserved N-terminal domain with a profilin-like fold called a longin domain (LD). These domains seem to be essential for regulating membrane trafficking and they mediate unexpected biochemical functions via a range of protein-protein and intramolecular binding specificities. In addition to the longins, proteins involved in the regulation of intracellular trafficking, such as subunits of the adaptor and transport protein particle complexes, also have LD like folds. The functions and cellular localization of longins are regulated at several levels and the longin prototypes TI-VAMP, Sec22 and Ykt6 show different distributions among eukaryotes, reflecting their modular and functional diversity. In mammals, TI-VAMP and Ykt6 are crucial for neuronal function, and defects in longin structure or function might underlie some human neurological pathologies. PMID- 15544956 TI - Preface to Peter Hochachka memorial volume. PMID- 15544957 TI - The great legacies of Peter Hochachka: a researcher, a teacher and a friend. PMID- 15544958 TI - Adaptation of enzymes to temperature: searching for basic "strategies". AB - The pervasive influence of temperature on biological systems necessitates a suite of temperature--compensatory adaptations that span all levels of biological organization--from behavior to fine-scale molecular structure. Beginning about 50 years ago, physiological studies conducted with whole organisms or isolated tissues, by such pioneers of comparative thermal physiology as V.Ya. Alexandrov, T.H. Bullock, F.E.J. Fry, H. Precht, C.L. Prosser, and P.F. Scholander, began to document in detail the abilities of ectothermic animals to sustain relatively similar rates of metabolic activity at widely different temperatures of adaptation or acclimation. These studies naturally led to investigation of the roles played by enzymatic proteins in metabolic temperature compensation. Peter Hochachka's laboratory became an epicenter of this new focus in comparative physiology. The studies of the enzyme lactate dehydrogenase (LDH) that he initiated as a PhD student at Duke University in the mid-1960s and continued for several years at the University of British Columbia laid much of the foundation for subsequent studies of protein adaptation to temperature. Studies of orthologs of LDH have revealed the importance of conserving kinetic properties (catalytic rate constants (kcat) and Michaelis-Menten constants (Km) and structural stability during adaptation to temperature, and recently have identified the types of amino acid substitutions causing this adaptive variation. The roles of pH and low-molecular-mass organic solutes (osmolytes) in conserving the functional and structural properties of enzymes also have been elucidated using LDH. These studies, begun in Peter Hochachka's laboratory almost 40 years ago, have been instrumental in the development of a conceptual framework for the study of biochemical adaptation, a field whose origin can be traced largely to his creative influences. This framework emphasizes the complementary roles of three "strategies" of adaptation: (1) changes in amino acid sequence that cause adaptive variation in the kinetic properties and stabilities of proteins, (2) shifts in concentrations of proteins, which are mediated through changes in gene expression and protein turnover; and (3) changes in the milieu in which proteins function, which conserve the intrinsic properties of proteins established by their primary structure and modulate protein activity in response to physiological needs. This theoretical framework has helped guide research in adaptational biochemistry for many years and now stands poised to play a critical role in the post-genomic era, as physiologists grapple with the challenge of integrating the wealth of new data on gene sequences (genome), gene expression (transcriptome and proteome), and metabolic profiles (metabolome) into a realistic physiological context that takes into account the evolutionary histories and environmental relationships of species. PMID- 15544959 TI - Hormones and fish hepatocyte metabolism: "the good, the bad and the ugly!". AB - This short review examines some of my personal experiences with Dr. Peter Hochachka, as a mentor and friend, and how his encouragement led to the research undertaken in my laboratory over the past three decades. Specifically, our work using the fish hepatocyte preparation as a model cell system is reviewed. The hepatocyte is an ideal cellular system that can be used to probe hepatic physiology and biochemistry. The impact of insulin, glucagon and related peptides, and catecholamines is discussed from the perspective of core and diverse functions of these key vertebrate metabolic hormones. Each hormone that operates in fish species was studied in manners similar to that of mammals, but it appears that the role of glucagon-like peptide-1 (GLP-1) in particular differs substantially from that in mammals. The receptors for each of these fish hormones seem structurally and in some cases functionally quite distinct from those in mammals. Few fish hormone receptor sequences are available, but fish genomists are rapidly adding new sequence information to the existing databases, so our view of the evolution of vertebrate hormone receptors will become clearer very quickly. PMID- 15544960 TI - Control of glycerol production by rainbow smelt (Osmerus mordax) to provide freeze resistance and allow foraging at low winter temperatures. AB - The rainbow smelt (Osmerus mordax) is a small anadromous fish that actively feeds under the ice at temperatures as low as the freeze point of seawater. Freezing is avoided through the production of both non-colligative antifreeze protein (AFP) and glycerol that acts in a colligative manner. Glycerol is constantly lost across the gills and skin, thus glycerol production must continue on a sustained basis at low winter temperatures. AFP begins to accumulate in early fall while water temperatures are still high. Glycerol production is triggered when water temperatures decrease to about 5 degrees C. Glycerol levels rapidly increase with carbon flow from dihydroxyacetone phosphate (DHAP) to glycerol 3-phosphate (G3P) to glycerol. Glucose/glycogen serves as the initial carbon source for glycerol accumulation with amino acids contributing thereafter. The period of glycerol accumulation is associated with increases in GPDH mRNA and PEPCK mRNA followed by elevations in protein synthesis and enzyme activities. Plasma glycerol levels may reach in excess of 500 mM in winter. The high freeze resistance allows rainbow smelt to invade water of low temperature and forage for food. The lower the temperature, the higher the glycerol must be, and the higher the glycerol the greater the loss to the environment through diffusion. During the winter, rainbow smelt feed upon protein rich invertebrates with glycerol production being fueled in part by dietary amino acids via the gluconeogenic pathway. At winter temperatures, glycerol is quantitatively more important than AFP in providing freeze resistance of blood; however, the importance of AFPs to other tissues is yet to be assessed. Glycerol levels rapidly plummet in the spring when water temperature is still close to 0 degrees C. During this period, freeze resistance must be provided by AFP alone. Overall, the phenomenon of glycerol production by rainbow smelt reveals an elegant connection of biochemistry to ecology that allows this species to exploit an otherwise unavailable food resource. PMID- 15544961 TI - Adventures in oxygen metabolism. AB - Peter W. Hochachka led a grand life of science adventure and left as his legacy a whole new field--biochemical adaptation. Oxygen was at the core of Peter's career and his laboratory made major contributions to our understanding of how animals deal with variation in oxygen availability in many forms. He analyzed the molecular mechanisms that support facultative anaerobiosis, studied muscle exercise metabolism for high speed flight, swimming and running, investigated mammalian diving on many trips to the Antarctic to study Weddell seals, and probed the metabolic and genetic adaptations that provide optimal hypoxia tolerance for humans residing at high altitudes. His work illuminated both biochemical and physiological mechanisms that are used to optimize aerobic metabolism, to compensate for hypoxic insults, and to conserve energy by strong metabolic rate depression under anoxia. His articles, books and lectures galvanized the field with leading-edge insights and theories and he consistently challenged comparative biochemists to use their unique model systems to explore the range and breadth of animal strategies of biochemical adaptation. Lessons drawn from my training in Peter's laboratory have led me on continuing explorations of adaptations in enzyme function, signal transduction, gene expression, and antioxidant defenses ranging over systems of anoxia tolerance, freezing survival, estivation, and mammalian hibernation. PMID- 15544962 TI - Locomotor performance and muscle metabolic capacities: impact of temperature and energetic status. AB - In aquatic ectotherms, muscle metabolic capacities are strongly influenced by exogenous factors, principally temperature and food availability. Seasonal changes in temperature lead many organisms to modify their metabolic machinery so as to maintain capacity even in "slower" cold habitats. Modifications of mitochondrial capacities are central in this response. The increases in protein specific oxidative capacities of mitochondria during cold acclimation of temperate fishes do not occur during the evolutionary adaptation to cold in Antarctic species. Instead, Antarctic fishes tend to increase the proportion of fibre volume devoted to mitochondria, perhaps to facilitate intracellular distribution of oxygen and metabolites. Variation in energetic status can drastically modify muscle metabolic status, with glycolytic muscle changing more than oxidative muscle. This in turn impacts swimming performance. A decrease in the condition of cod leads endurance at speeds above Ucrit to drop by 70%. Sprint swimming is less affected, perhaps as it does not exhaust glycolytic muscle. We used interindividual variation in muscle metabolic capacities to identify correlates of swimming performance in stickleback and cod. Activities of cytochrome c oxidase in glycolytic muscle are a correlate of sprint swimming in stickleback (Gasterosteus aculeatus) and cod (Gadus morhua), whereas lactate dehydrogenase activities in glycolytic muscle are a correlate of cod endurance swimming. In scallops, gonadal maturation leads to virtually complete mobilisation of glycogen from muscle. This does not reduce the capacity of the scallops, Chlamys islandica and Euvola ziczac, to mount escape responses, but significantly slows their recuperation from exhaustive exercise. Muscle metabolic capacities fall in parallel with glycogen mobilisation. In the compromise between muscles' dual roles as a motor and a macromolecular reserve, a significant loss in locomotory ability occurs during gametogenesis and spawning. Reproductive fitness takes the upper hand over maintenance of performance. PMID- 15544963 TI - Salmon spawning migration and muscle protein metabolism: the August Krogh principle at work. AB - The August Krogh principle, stating that for any particular question in biology, nature holds an ideal study system, was applied by choosing the anorexic, long distance migration of salmon as a model to analyze protein degradation and amino acid metabolism. Reexamining an original study done over 20 years ago on migrating sockeye salmon (Oncorhynchus nerka), data on fish migration and starvation are reviewed and a general model is developed on how fish deal with muscle proteolysis. It is shown that lysosomal activation and degradation of muscle protein by lysosomal cathepsins, especially cathepsin D and sometimes cathepsin L, are responsible for the degradation of muscle protein during fish migration, maturation and starvation. This strategy is quite the opposite to mammalian muscle wasting, including starvation, uremia, cancer and others, where the ATP-ubiquitin proteasome in conjunction with ancillary systems, constitutes the overwhelming pathway for protein degradation in muscle. In mammals, the lysosome plays a bit part, if any. In contrast, the proteasome plays at best a subordinate role in muscle degradation in piscine systems. This diverging strategy is put into the context of fish metabolism in general, with its high amino acid turnover, reliance on amino acids as oxidative substrates and flux of amino acids from muscle via the liver into gonads during maturation. Brief focus is placed on structure, function and evolution of the key player in fishes: cathepsin D. The gene structure of piscine cathepsin D is outlined, focusing on the existence of duplicate, paralogous, cathepsin D genes in some species and analyzing the relationship between a female and liver-specific aspartyl protease and fish cathepsin Ds. Evolutionary relationships are developed between different groups of piscine cathepsins, aspartyl proteases and other cathepsins. Finally, based on specific changes in muscle enzymes in fish, including migrating salmon, common strategies of amino acid and carbon flux in fish muscle are pointed out, predicting some metabolic concepts that would make ideal application grounds for the August Krogh principle. PMID- 15544964 TI - Adenosine as a signal for ion channel arrest in anoxia-tolerant organisms. AB - Certain freshwater turtles and fish are extremely anoxia-tolerant, capable of surviving hours of anoxia at high temperatures and weeks to months at low temperatures. There is great interest in understanding the cellular mechanisms underlying anoxia-tolerance in these groups because they are anoxia-tolerant vertebrates and because of the far-reaching medical benefits that would be gained. It has become clear that a pre-condition of prolonged anoxic survival must involve the matching of ATP production with ATP utilization to maintain stable ATP levels during anoxia. In most vertebrates, anoxia leads to a severe decrease in ATP production without a concomitant reduction in utilization, which inevitably leads to the catastrophic events associated with cell death or necrosis. Anoxia-tolerant organisms do not increase ATP production when faced with anoxia, but rather decrease utilization to a level that can be met by anaerobic glycolysis alone. Protein synthesis and ion movement across the plasma membrane are the two main targets of regulatory processes that reduce ATP utilization and promote anoxic survival. However, the oxygen sensing and biochemical signaling mechanisms that achieve a coordinated reduction in ATP production and utilization remain unclear. One candidate-signaling compound whose extracellular concentration increases in concert with decreasing oxygen availability is adenosine. Adenosine is known to have profound effects on various aspects of tissue metabolism, including protein synthesis, ion pumping and permeability of ion channels. In this review, I will investigate the role of adenosine in the naturally anoxia-tolerant freshwater turtle and goldfish and give an overview of pathways by which adenosine concentrations are regulated. PMID- 15544965 TI - Hochachka's "Hypoxia Defense Strategies" and the development of the pathway for oxygen. AB - Hochachka's "Hypoxia Defense Strategies" identify oxygen signalling, metabolic arrest, channel arrest and coordinated suppression of ATP turnover rates as key factors that determine the ability of organisms to survive exposure to chronic hypoxia. In this review, I assess the developmental role played by these phenomena in the morphogenesis of the gas exchange tissues that define the pathway for oxygen transport to cytochrome c oxidase. Key areas of regulation lie in: (I) the suppression of fetal mitochondrial oxidative function in hand with mitochondrial biogenesis (metabolic arrest), (II) the role of hypoxia-driven oxygen signalling pathways in directing the scope of non-differentiated stem cell proliferation in placenta and lung development and (III) the regulation of epithelial fluid secretion/absorption in the lung through the oxygen-dependent modulation of Na+ conductance pathways. The identification of developmental roles for Hochachka's "Hypoxia Defense Strategies" in directing the morphogenesis of gas exchange structures bears with it the implication that these strategies are fundamental to establishing the scope for aerobic metabolic performance throughout life. PMID- 15544966 TI - The biochemistry of metabolic depression: a history of perceptions. AB - Our interest in animals that 'turn off' dates back at least 300 years. This phenomenon has been reported in most of the major invertebrate phyla and in all vertebrate classes, and has implications for our understanding of a wide range of homeostasis and metabolic control issues. Surprisingly however, it took 20 years of biochemical research before the realization dawned that metabolic depression is the frontline strategy utilized by these animals to survive environmental stress. In this essay, the history of this research is treated in five stages, defined in terms of how the phenomenon now known as metabolic depression was perceived at the time. The two initial stages clearly show that the researchers involved were refractory to the concept of metabolic depression until about 1982 (stage 3). The two stages after 1982 reflect the impact of the acknowledgement of metabolic depression per se and show how research is now being directed towards both the mechanisms involved in, and the cellular targets of metabolic depression. PMID- 15544967 TI - Fat to the fire: the regulation of lipid oxidation with exercise and environmental stress. AB - Lipids are an important fuel for submaximal aerobic exercise. The ways in which lipid oxidation is regulated during locomotion is an area of active investigation. Indeed, the integration between cellular regulation of lipid metabolism and whole-body exercise performance is a fascinating but often overlooked research area. Additionally, the interaction between environmental stress, exercise, and lipid oxidation has not been sufficiently examined. There are many functional and structural steps as fatty acids are mobilized, transported, and oxidized in working muscle, which may serve either as regulatory points for responding to acute or chronic stimuli or as raw material for natural selection. At the whole-animal level, the partitioning of lipids and carbohydrates across exercise intensities is remarkably similar among mammals, which suggests that there is conservation in regulatory mechanisms. Conversely, the proportions of circulatory and intramuscular fuels differ between species and across exercise intensities. Responses to acute and chronic environmental stress likely involve the interaction of genetic and nongenetic changes in the fatty acid pathway. Determining which of these factors help regulate the fatty acid pathway and what impact they have on whole-animal lipid oxidation and performance is an important area of future research. Using an integrative approach to complete the information loop from gene to physiological function provides the most powerful mode of analysis. PMID- 15544968 TI - Mitochondria: aerobic and anaerobic design--lessons from molluscs and fishes. AB - The contributions of Peter Hochachka to the development of comparative and adaptational biochemistry are substantial. In particular, he and his academic offspring made major contributions to the understanding of the metabolism of molluscs and fishes. These two large taxonomic groups each have marine, freshwater and terrestrial/semiterrestrial representatives, and their mitochondrial metabolism has been shaped by these environmental conditions. In particular, the importance of amino acids and lipids as energy sources has interesting correlations with the environment and the osmotic strategy used. In marine molluscs, amino acids are important aerobic energy sources, and are used as osmolytes and participate in anaerobic metabolism. In marine elasmobranchs, amino acids and ketone bodies, but not lipids per se, are important energy sources in extrahepatic tissues. Marine and freshwater teleost fish by contrast use lipids as an extrahepatic energy source with minimal use of ketone bodies. Furthermore, ketone bodies are important in the metabolism of freshwater and terrestrial but not marine molluscs. The bases for these different metabolic plans may lie in the solute systems used by the different groups (e.g. amino acids in marine molluscs and urea in marine elasmobranchs). The various metabolic options used by fishes and molluscs indicate the plasticity of metabolic design in an environmental context. PMID- 15544969 TI - Organ arrest, protection and preservation: natural hibernation to cardiac surgery. AB - Cardiac surgery continues to be limited by an inability to achieve complete myocardial protection from ischemia-reperfusion injury. This paper considers the following questions: (1) what lessons can be learned from mammalian hibernators to improve current methods of human myocardial arrest, protection and preservation? and (2) can the human heart be pharmacologically manipulated during acute global ischemia to act more like the heart of a hibernating mammal? After reviewing the major entropy-slowing strategies of hibernation, a major player identified in the armortarium is maintenance of the membrane potential. The resting membrane potential of the hibernator's heart appears to be maintained close to its pre-torpid state of around -85 mV. In open-heart surgery, 99% of all surgical heart arrest solutions (cardioplegia) employ high potassium (>16 mM) which depolarises the membrane voltage from -85 to around -50 mV. However, depolarising potassium cardioplegia has been increasingly linked to myocyte and microvascular damage leading to functional loss during reperfusion. Our recent work has been borrowed from hibernation biology and is focused on a very different arrest strategy which 'clamps' the membrane near its resting potential and depresses O2 consumption from baseline by about 90%. The new 'polarising' cardioplegia incorporates adenosine and lidocaine (AL) as the arresting combination, not high potassium. Studies in the isolated rat heart show that AL cardioplegia delivered at 37 degrees C can arrest the heart for up to 4 h with 70 80% recovery of the cardiac output, 85-100% recovery of heart rate, systolic pressure and rate-pressure product and 70-80% of baseline coronary flows. Only 14% of hearts arrested with crystalloid St. Thomas' solution No. 2 cardioplegia survived after 4 h. In conclusion, maintenance of the myocardial membrane potential near or close to its resting state appears to be an important feature of the hibernator's heart that may find great utility in surgical arrest and cellular preservation strategies. Identifying and safely turning 'off' and 'on' the entropy-slowing genes to down-regulate the hibernator's heart and applying this to human organs and tissues remains a major challenge for future genomics and proteomics. PMID- 15544971 TI - Regulation of malignant progression by the hypoxia-sensitive transcription factors HIF-1alpha and MTF-1. AB - Solid tumors are known to develop microenvironmental hypoxia or anoxia due to malfunction and malformation of blood vessels and the energy demands of the highly proliferative tumor cells. Oxygen deprivation can cause aberrant modifications of signaling pathways and their downstream transcription factors that are believed to contribute to malignancy. Here, we review the latest studies related to the involvement of hypoxia-inducible transcription factor-1alpha (HIF 1alpha), the first known mammalian intracellular hypoxia sensor, in tumor development. We propose that a second far less studied protein, metal transcription factor-1 (MTF-1), acts as a more general oxygen sensor, responding to both hypoxia and oxidative stress, and is also intimately involved in malignant progression. Existing evidence suggests that activation of these two ubiquitous proteins, by hypoxia and genetic modifications, modulate the expression patterns of a number of important proteins involved in tumorigenesis. PMID- 15544970 TI - Using humans to study the physiological evolution of energy metabolism: a tribute to Peter Hochachka. AB - Comparative physiology traditionally relies on unusual animals to explore the mechanistic basis or evolution of physiological processes. Despite his credentials as a comparative physiologist, much of the recent work by Peter Hochachka relied upon humans as "comparative" models. Technological advances in clinical analyses and expanding databases of genomic diversity have made humans a very "convenient" model to study the evolution of metabolism. For example, a growing number of metabolic diseases have been shown to have a genetic basis. Obesity, type II diabetes, metabolic syndrome X may each have linkages to polymorphisms in genes encoding transcriptional regulators of bioenergetic genes. Genetic polymorphisms and gene families for these important transcription factors inspire questions about the evolution of energy metabolism in other animals. Thus, these biomedical studies provide important direction for researchers interested in exploring the regulatory basis of interspecies variations in bioenergetics. PMID- 15544972 TI - Defining the limits of diving biochemistry in marine mammals. AB - The field of marine mammal diving biochemistry was essentially untouched when Peter Hochachka turned his attention to it in the mid-1970s. Over the next 30 years, his work followed three main themes in this area: first, most biologists at that time supported the theory that diving mammals utilized enhanced metabolic pathways for hypoxic energy production (glycolysis to lactate) and reduced their metabolic rate while diving. Peter began his work on potential hypoxic adaptations in marine mammals by working out the details of how these pathways would be regulated. By the 1980s, he started to ask how diving mammals balanced the increased demands of exercise with the apparently conflicting demands to reduce aerobic metabolism while exercising underwater. By the 1990s, his work involved complex models of the interplay between the neural, hormonal, behavioral and evolutionary components of diving biochemistry and animal exercise. From a comparative approach, he excelled at bringing themes of hypoxic adaptation from many different types of animals to the field of diving mammal biochemistry. This review traces the history of Peter Hochachka's work on diving biochemistry from the perspective of those of us who spent time with him both inside the laboratory and outside in the field from Antarctica to Iceland. PMID- 15544973 TI - Changes in gene expression as biochemical adaptations to environmental change: a tribute to Peter Hochachka. AB - Changes in gene expression are likely to play a critical role in both acclimation and adaptation to a changing environment. There is a rapidly growing body of literature implicating quantitative changes in gene expression during acclimation to environmental change, but less is known about the role of qualitative changes in gene expression, such as switching between alternative isoforms. Alternative isoforms can arise via gene duplication, alternative splicing, or alternative promoter usage. Organisms that have undergone recent genome duplication events may make use of environment-specific isoforms coded by multiple genes, but their role in other organisms is less well known. However, recent data suggest that isoforms arising from alternative splicing may be an under-appreciated source of physiological variation. The role of changes in gene expression during evolutionary adaptation has received comparatively limited attention, but novel approaches to addressing the adaptive significance of changes in gene expression have been applied to a few cases of differences in gene expression among taxa. Recent advances in genomics, including microarray technology, knock-out and knock down approaches, and the wealth of data coming from large-scale sequencing projects have provided (and will continue to provide at ever increasing rates) new insights into these classic questions in comparative biochemistry. PMID- 15544974 TI - Metabolic scaling: a many-splendoured thing. AB - Animals at rest and during exercise display rates of aerobic metabolism, VO2, that represent mainly the sum of mitochondrial respiration rates in various organs. The relative contributions of these organs change with physiological state such that internal organs such as liver, kidney and brain account for most of the whole-body VO2 at rest, while locomotory muscles account for >90% of the maximum rate, VO2max, during maximal aerobic exercise. Mechanisms that regulate VO2 are complex and the relative importance of each step in a series, estimated by metabolic control analysis, depends upon the level of biological organization under consideration as well as physiological state. Despite this complexity, prominent single-cause models propose that metabolic rates are supply-limited and that the scaling of supply systems provides a sufficient explanation for the allometric scaling of metabolism. We argue that some assumptions, as well as current interpretations of the meaning (or consequences) of these constraints are flawed, i.e., elephants do not have lower mass-specific basal or maximal rates of aerobic metabolism because their mitochondria are more supply-limited than those of shrews. Animals do not violate the laws of physics, and the allometric scaling of supply systems would be expected, to some extent, to be matched by capacities for (and rates of) energy expenditure. But life is not so simple. Animals are so diverse that to do justice to metabolic scaling, it is also necessary to consider the scaling of energy expenditure. It is by doing so that models of metabolic scaling can be consistent with current paradigms in metabolic regulation and accommodate the range of inter- and intraspecific exponents found in nature. The "allometric cascade," a first attempt at such an accounting, was a source of great satisfaction to Peter Hochachka. It was the last door that he helped open to comparative physiologists before he said goodbye. PMID- 15544975 TI - The definition and assessment of pain. PMID- 15544976 TI - Pain in women's health: a multi-faceted approach toward understanding. AB - Pain has always been a part of women's health experiences, inherent to such physiologic processes as menstrual cramping, labor contractions, and uniquely female illnesses, such as cervical or ovarian cancer. However, the understanding of pain-its nature, its purpose, and its sometimes debatable need for removal remains elusive. Pain's origins are in the physical realm, but it is manifested through an array of psychological, social, and cultural factors. The concept of pain is explored using an evolutionary approach to understanding the mechanisms associated with the physiologic, psychological, developmental, and sociocultural aspects of this phenomenon. The relevance of this exploratory look at pain as it relates to offering health care to women is discussed. The manner in which pain affects individuals and the methods with which it can be treated are critical elements in the provision of quality health care. PMID- 15544977 TI - The neuromatrix theory of pain: implications for selected nonpharmacologic methods of pain relief for labor. AB - Women experience the pain of labor differently, with many factors contributing to their overall perception of pain. The neuromatrix theory of pain provides a framework that may explain why selected nonpharmacologic methods of pain relief can be quite effective for the relief of pain for the laboring woman. The concept of a pain "neuromatrix" suggests that perception of pain is simultaneously modulated by multiple influences. The theory was developed by Ronald Melzack and represents an expansion beyond his original "gate theory" of pain, first proposed in 1965 with P. D. Wall. This article reviews several nonpharmacologic methods of pain relief with implications for the practicing clinician. Providing adequate pain relief during labor and birth is an important component of caring for women during labor and birth. PMID- 15544978 TI - Update on nonpharmacologic approaches to relieve labor pain and prevent suffering. AB - The control of labor pain and prevention of suffering are major concerns of clinicians and their clients. Nonpharmacologic approaches toward these goals are consistent with midwifery management and the choices of many women. We undertook a literature search of scientific articles cataloged in CINAHL, PUBMED, the Cochrane Library, and AMED databases relating to the effectiveness of 13 non pharmacologic methods used to relieve pain and reduce suffering in labor. Suffering, which is different from pain, is not an outcome that is usually measured after childbirth. We assumed that suffering is unlikely if indicators of satisfaction were positive after childbirth. Adequate evidence of benefit in reducing pain exists for continuous labor support, baths, intradermal water blocks, and maternal movement and positioning. Acupuncture, massage, transcutaneous electrical nerve stimulation, and hypnosis are promising, but they require further study. The effectiveness of childbirth education, relaxation and breathing, heat and cold, acupressure, hypnosis, aromatherapy, music, and audioanalgesia are either inadequately studied or findings are too variable to draw conclusions on effectiveness. All the methods studied had evidence of widespread satisfaction among a majority of users. PMID- 15544979 TI - Obstetric anesthesia: changes and choices. AB - This article is a review of analgesics and anesthetics offered to laboring women, including intravenous drugs, epidural and spinal agents, and inhalational anesthetics. An overview of the uses, risks, and benefits is provided for each anesthetic alternative. To provide the most effective care to women in labor, clinicians have a responsibility to have current knowledge of the best evidence for safety and efficacy of these pharmacologic agents and techniques and be able to effectively communicate this information to clients. PMID- 15544980 TI - Pain assessment and intervention for term newborns. AB - A baseline assessment of pain should occur as part of a newborn's health assessment within the first few hours of life. Validation and assessment of pain in the term newborn is important for providing care because newborns may experience pain from a multitude of procedures, ranging from heel sticks to circumcisions. Current assessment tools evaluate both behavioral and physiologic parameters. In addition, providing individualized care requires a knowledge base of the interventions available to reduce or eliminate pain, such as breastfeeding, non-nutritive suck, skin-to-skin contact, and sucrose pacifiers. This article describes three newborn pain assessment tools, the use of valid assessment tools, and evidence-based interventions that are recommended to effectively manage newborn pain. PMID- 15544981 TI - Primary dysmenorrhea: assessment and management update. AB - Many women are familiar with the experience of dysmenorrhea, which can contribute to significant physical and emotional distress and life disruption. However, women may not seek professional expertise in their attempt to alleviate this condition. It is important to assess the beliefs and experiences of all women with dysmenorrhea, including adolescents, as early in gynecologic care as possible. This article reviews the management of primary dysmenorrhea. Midwives can provide valuable assistance to women in their explorations of the variety of treatment options available for the relief of dysmenorrhea, including lifestyle changes, complementary and alternative approaches, analgesics, and hormones. PMID- 15544982 TI - Pain at midlife. AB - Although menopause is a normal developmental milestone through which all women pass, the transition has been long associated with chronic pain conditions that may be more accurately viewed as secondary to aging. Clinicians need to understand management of pain problems women may experience. This article examines pain syndromes including headache, back pain, osteoarthritis, pelvic pain, vulvo-vaginal pain, and burning mouth syndrome. PMID- 15544983 TI - Management of ruptured membranes at term. PMID- 15544984 TI - The World Health Organization analgesic ladder. PMID- 15544988 TI - Female companionship during labor: a crucial resource in times of stress. PMID- 15544989 TI - Dissociation and childhood abuse history in epileptic and pseudoseizure patients. AB - The aim of this study was to examine dissociative experiences, childhood abuse and anxiety in epileptic and pseudoseizure female patients. Thirty-three patients with pseudoseizures and thirty patients with epilepsy were recruited from Cumhuriyet University Hospital Psychiatry and Neurology Units. We assessed each participant using the Dissociative Experiences Scale, the Clinician-Administered Dissociative States Scale and the Childhood Abuse and Neglect Questionnaire. The female patients with pseudoseizures showed significantly higher levels of dissociative experiences and childhood trauma. Epileptic female patients showed higher levels of anxiety. The significantly higher incidence of dissociative experiences and childhood trauma in the patients with pseudoseizures makes a case for dissociation in the pathogenesis of these seizures. PMID- 15544990 TI - The role of PET in presurgical assessment of partial epilepsies. AB - In the presurgical assessment of drug refractory partial epilepsies PET represents a privileged means for investigating glucose metabolism and neurotransmission in the functional-deficit zone, defined as the region of cortex that is functioning abnormally in the interictal period. Another aim of PET investigation is to produce images of neurotransmission abnormalities underlying neuronal hyperexcitability, and thus to allow direct visualization of the epileptogenic zone. This approach has been mostly based on the binding of various radio-ligands on specific receptors (GABA-A, opiate and serotonin receptor) and on brain uptake of serotonin precursors. These attempts have not yet been fully validated, in spite of promising studies of serotonin synthesis and receptors. Consequently, at the present state of their development, most of the PET techniques routinely used, reflect changes that are not directly related to the epileptogenic process itself. The lack of large multicentric controlled studies, evaluating the impact of PET, represents the main limitation to a better understanding of the clinical role and utility of PET in epilepsy. We review the basic aspects and limitations of the technique, the various radiopharmaceuticals that have been tested in epilepsy, the sensitivity of the different types of PET investigations, the pathophysiology of PET abnormalities and discuss the practical utility of PET imaging in presurgical assessment of partial epilepsies. PMID- 15544991 TI - [Microenvironment and tumor progression: the progress concept]. PMID- 15544992 TI - [Burn out syndrome in oncology]. AB - SEPS or burnout syndrome was described among health care workers. Oncology care givers--physicians and nurses--can be concerned. Burnout is a chronical stress reaction. Emotional exhaustion and depersonalisation are more prevalent than low personal accomplishment. Burnout is essentially assessed by questionnaires. Oncologists report an higher level of burnout, than AIDS medical or palliative care staff. Causes of burn out are numerous: insufficient personal time, sense of failure,... followed by poorly management and difficulties in staff or institution relationships. Prevention and therapy of burn out can be considered on three levels: personal, (psychotherapy, advices on health way of life), team (improvement in communication) and institution (support meetings and talking groups). PMID- 15544993 TI - [Hypoxia, HIF1alpha and estrogen receptor]. AB - The estrogen receptor plays an important role in the development of hormone dependent breast cancer. Experimental and clinical studies showed that hypoxia was involved in tumor progression and in the resistance to anticancer agents. The scope of this review is to summarize recent observations on these two signaling pathways and to show their importance and possible interaction in the development of hormone-dependent breast cancer. PMID- 15544994 TI - [Targeting of membrane receptor tyrosine kinases: is there resistance in the HER?]. AB - Human Epidermal growth factor Receptors (HER) play an important role in cellular proliferation, and differentiation. Their overexpression in tumor tissues is often associated with a poor prognosis. Consequently, HER receptors are interesting therapeutic targets for cancer treatment. Two strategies are proposed. First, monoclonal antibodies can be used to inhibit the binding of one ligand to its receptor. The second approach is based upon the designing of tyrosine kinase inhibitors capable to bind into the phosphorylation site of the receptor. Consequently, both approaches block the signal transduction downstream. Resistance to anti receptor tyrosine kinase therapy can lead to enhanced morbidity associated with high therapeutic cost. Different mechanisms can be implicated. Non specific mechanisms include alterations of the signal transduction pathways (PI3K/AKT), recruitment of alternative receptor tyrosine kinase pathways (IGFR, VEGFR) and proteasome degradation inhibition. Other mechanisms are specific to HER and rely on inhibition of the binding of monoclonal antibodies (sialomucin-MUC4), heterodimerisation of HER, truncated soluble receptors intervention and mutated variants, as demonstrated very recently with EGF receptors, or genetic polymorphism. This paper reviews these different resistance mechanisms that have been identified in preclinical and clinical situations. PMID- 15544995 TI - [Fluorinated analogs of nucleosides and fluorinated tracers of gene expression for positron emission tomography]. AB - 18F-FDG is currently the only fluorinated tracer used in routine clinical positron emission tomography (PET). Fluorine 18 is considered as the ideal radioisotope for PET, thanks to a low positron energy, which not only limits the dose rate to the patients but also provides high-resolution images. Furthermore, the 110 min. physical half-life allows for high-yield radiosynthesis, transport from the production site to the imaging site, and imaging protocols that could span hours, which permits dynamic studies and assessing metabolic processes that may be fairly slow. Recently, synthesis of fluorinated tracers from prosthetic group precursors, which allows easier radiolabeling of biomolecules, has given a boost to the development of numerous fluorinated tracers. Given the wide availability of fluorine 18, such tracers may well develop into important routine tracers. This article is a review of the literature concerning fluorinated analogs of nucleosides and fluorinated radiotracers of gene expression recently developed and under investigation. PMID- 15544996 TI - [To stimulate or to inhibit nitric oxide production in mammary tumors?]. AB - NO is a molecule produced in different amounts by two types of enzymes, constitutive NO-synthases and inducible NO-synthase, the last one produce large amount of NO. In tumor outcome, its role may be either beneficial or detrimental due to its actions in the different steps of tumor growth and metastasis. This review deals with mammary tumors and the mechanisms lying behind NO effects. In human patients, increased amounts of NO were evidenced in blood, linked with the presence and activity of NO-synthase in breast tumors. Non-unequivocal correlations were established with tumor grade, invasiveness and metastatic potential. Studies in animal models have given hints to explain these discrepancies by the type of the involved NO-synthase, the amount of NO it produces, and its belonging to tumoral or stromal cells. Indeed, it was recently shown that NO produced by tumor cells inhibits, while NO produced by stromal cells facilitates tumor growth, at least in the model which was studied. On the one hand, NO toxicity against tumor cells is a well known phenomenon, but on the other hand, NO may promote tumor invasiveness due to its effect on extracellular matrix, and to its angiogenetic properties. So the role of NO in mammary tumor outcome is not clear-cut, and, at the present time, it cannot be ascribed a pronostic value in breast tumor. However, researches aimed at managing tumor cells to produce NO sufficient to induce their death may be fruitful since, be tumor targeting successful, no general toxicity would be encountered. PMID- 15544997 TI - [Acute lymphoblastic leukemia in elderly: prognosis and treatment]. AB - Over 60 years old, acute lymphoblastic leukemia (ALL) represents between 16 and 31% of all adult cases. Pre-B and common ALL are frequent, while T-cell lineage ALL is under-represented in elderly populations as compared with younger adults. The frequency of Philadelphia chromosome seems also to increase with age and adversely influences complete remission (CR) and survival rates. Poor performance status, co-morbidity factors and early mortality during intensive induction chemotherapy are the main reasons for poor outcome. Few reports on effectiveness and toxicity of therapeutic regimens involving exclusively elderly patients with ALL have been published and only some of them were prospective studies. Age adapted approaches with less aggressive chemotherapy have been applied. The overall response ranged from 12 to 85%. Toxic death during induction chemotherapy was observed in 7 to 42% of the patients. Among the patients who received a curative approach, the median overall survival duration ranged from 3 to 14 months, while it ranged from 1 to 14 months for those treated palliatively. New therapeutic approaches are warranted to improve the outcome in this age group of ALL patients. PMID- 15544998 TI - [MRI evaluation of primary chemotherapy response in breast cancer]. AB - The aim of this work was to evaluate the value of contrast enhanced MRI for determination of response to neoadjuvant chemotherapy (type FEC) in breast cancer according to two parameters: size of the enhancing tumor and the maximum relative enhancement curve (MRC) in the same tumor area. Twenty women with breast cancer (15 invasive ductal carcinomas and 5 invasive lobular carcinomas) T2 (n = 8) or T3 (n = 12) were evaluated by physical examination and MRI after a minimal of three courses of FEC and prior to surgery. Data from physical examination and imaging studies were compared to histopathological findings. Physical examination estimated correctly the residual tumor size in 45% of cases and MRI in 60% with 3 false negative cases. Among evaluated patients with MRI measurable residual tumor, tumor size was underestimated in 69% of the cases and overestimated in 31% of the cases. A MRC flattening was observed in 5 cases among the patients with a partial response or clinical stable disease correlated with a poor cellular density in the microscopic findings. MRI monitoring of chemotherapy response can be useful for guiding surgery. Therefore, underestimation of the residual tumor size and false negative rate are remaining problems. PMID- 15544999 TI - [Economic impact of outpatient chemotherapy in multiple myeloma]. AB - The development of alternatives to hospitalisation is mandatory in cancer patients. A retrospective analysis compared the costs relative to two modalities of administration of VAD chemotherapy (hospital versus outpatient) in multiple myeloma. This study was carried out from the perspective of the French health care system, including direct hospital and ambulatory costs. The robustness of results was tested in a sensitivity analysis. Forty four patients were included: 27 in the "hospitalisation" arm and 17 in the "outpatient" arm. Among the cycles planned to be administered in outpatient, only 5 cycles (4 patients) have to be administered in hospital. The length of hospitalisation was consequently reduced with outpatient administration (-11.2 days) and it induced a significant cost saving as 3,066 euros (-32%) per patient. In sensitivity analysis, this cost saving varied from 1,282 to 3,667 euros. Our study showed the safety and the economic interest of outpatient administration of VAD chemotherapy in multiple myeloma. Facilities and training are required to develop ambulatory chemotherapy. PMID- 15545000 TI - An ethics dilemma: when parents and doctors disagree on the best treatment for the child. AB - The increasing complexity of present day medicine--with highly effective and yet risky treatments, individual and collective expectations, and evolving ideological and cultural landmarks--often gives rise to difficult ethical problems. Specific meetings are valuable for understanding such problems, acquiring the relevant skills and for gaining and transmitting experience on how to solve them. Parents and doctors may disagree about what is the best treatment. Such a difference of opinion is not rare but usually a solution can easily be found. This is not the case when the child is treated for a severe illness and when there is no clearly defined or satisfactory treatment for him?her. We present how a dramatic conflict arose between the parents and the doctors faced with such a case (mostly because the staff failed to understand early enough the psychological factors at the root of the father's demands), how clinical, institutional and ethical problems were analysed during a meeting, and how they were solved. PMID- 15545001 TI - Taurine inhibition of metal-stimulated catecholamine oxidation. AB - Taurine is an abundant amino acid found in mammalian tissues and it has been suggested to have cytoprotective functions. The aim of the present study was to determine if taurine had the potential to reduce oxidative stress associated with metal-stimulated catecholamine oxidation. Taurine and structural analogs of taurine were tested for their ability to inhibit metal-stimulated quinone formation from dopamine or L-dopa. Oxidative damage to proteins and lipids were also assessed in vitro and the effects of taurine were determined. Taurine (20 mM) was found to decrease significantly ferric iron (50-500 microM)- and manganese (10 microM)-stimulated L-dopa or dopamine oxidation. Taurine had no effect on zinc-induced dopamine oxidation and slightly potentiated copper- and NaIO(4)-stimulated quinone formation. Ferric iron-stimulated lipid peroxidation was not affected by taurine (1-20 mM). Protein carbonyl formation induced by ferric iron (500 microM) and L-dopa (500 microM) was significantly reduced by 10 mM taurine. The cytotoxicity of L-dopa (250 microM) and ferric chloride (75 microM) to LLC-PK(1) cells was attenuated by 10 mM taurine or hypotaurine. Homotaurine alone stimulated L-dopa oxidation and potentiated the cytotoxic effects of ferric iron. Homotaurine was found to be cytotoxic when combined with L-dopa or L-dopa/iron. In contrast, hypotaurine inhibited quinone formation and protected LLC-PK(1) cells. These studies suggest that taurine may exhibit cytoprotective effects against the oxidation products of catecholamines by acting as a scavenger for free radicals and cytotoxic quinones. PMID- 15545002 TI - Effects of imidazenil, a new benzodiazepine receptor partial agonist, in the treatment of convulsions in organophosphate intoxications. AB - Anticonvulsant and antilethal effects of imidazenil, a new imidazobenzodiazepine derivative, in fluostigmine (DFP; diisopropyl phosphorofluoridate) intoxications were studied and compared with the effects of diazepam on mice and rats. Special attention was payed to the myorelaxant effects of both drugs. It was stated that imidazenil (i) significantly decreased convulsion intensity in mice, (ii) quickly inhibited seizure patterns in bioelectrical activity in the rat's brain, (iii) significantly increased antilethal effectiveness of the standard therapy in mice intoxicated with DFP. These effects are comparable to those of diazepam. However, effects of imidazenil in the rota-rod test of the mouse were noted in doses 5-10 times higher than therapeutic ones, when effects of diazepam on motor co ordination were seen in therapeutic dosage. PMID- 15545003 TI - Immediate early gene expression in rat basal ganglia after destruction of the dopaminergic system. AB - In the present study, we performed immunocytochemical mapping of cFos and c-Jun as markers for changes in neural activity in the brains of dopamine denervated rats. We observed a prolonged c-Fos and c-Jun expression in basal ganglia motor and limbic circuits over 96 h. This might be due to alterations in transmitter balances. We conclude, that changes in neural activities are involved in the development of 'extranigral' pathology of Parkinson's disease. PMID- 15545004 TI - Neurochemical and behavioural correlates in cassava-induced neurotoxicity in rats. AB - Chronic cyanide intoxication from cassava has been implicated as the cause for a degenerative neuropathy known widely as tropical ataxic neuropathy. An attempt has been made in this study to identify the specific cause for neuropathy caused by cassava using Wistar strain albino rats as the experimental animal model. The results of cassava fed animals were compared with control animals, animals given cyanide, malnourished animals and malnourished animals fed cyanide, to identify the causative factors. This study revealed that though the behavioural pattern in motor coordination of the cassava fed animals was similar to the other groups studied, the neurochemical basis for the observed behavioural pattern was unique for cassava. Hence the neurotoxicity of cassava could be attributed to unmetabolized linamarin, more than its nutritional status and/or cyanide toxicity. PMID- 15545005 TI - Establishment of an in vitro screening model for neurodegeneration induced by antimalarial drugs of the artemisinin-type.. AB - The establishment of an in vitro screening model for neurodegeneration inducing antimalarial drugs was conducted in stepwise fashion. Firstly, the in vivo selective neurotoxic potency of artemisinin was tested in neuronal cells in vitro in relation to the cytotoxic potency in other organ cell cultures such as liver and kidney or versus glial cells. Secondly, a comparison between different parts of the brain (cortex vs. brain stem) was performed and in the last step, a fast and sensitive screening endpoint was identified. In summary, non-neuronal cell lines such as hepatocytes (HEP-G2), liver epithelial cells (IAR), proximal tubular cells (LLC-PK(1)) and glial cells from the rat (C6) and human (GO-G-IJKT) displayed only moderate sensitivity to artemisinin and its derivatives. The same was found in undifferentiated neuronal cell lines from the mouse (N-18) and from human (Kelly), whereas during differentiation, these cells became much more sensitive. Primary astrocytes from the rat also were not specifically involved. In the comparison of primary neuronal cell cultures from the cortex and brain stem of the rat, the brain stem was found to be more sensitive than the cortex. The neurotoxic potential was determined by cytoskeleton elements (neurofilaments), which were degradated in vitro by diverse neurodegenerative compounds. In comparison of dog and rat primary brain stem cultures, the dog cells were found to be more sensitive to artemisinin than the rat cells. In addition to the primary brain stem cell cultures it was shown that the sprouting assay, which determines persistent delayed neurotoxic effects, is also useful for screening antimalarial drugs. To other compounds, artemether and artesunate, showed that use of the sprouting assay followed by primary brain stem cultures of the rat will be a good strategy to select candidate compounds. PMID- 15545006 TI - AMPA prevents glutamate-induced neurotoxicity and apoptosis in cultured cerebellar granule cell neurons. AB - Exposure of cultured cerebellar neurons to alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) in the presence of aniracetam protects all of the vulnerable neurons against the excitotoxic actions of glutamate acting on N methyl-D-aspartate receptors. The protective effect of AMPA was both time- and concentration-dependent. Aniracetam alone did not protect the neurons against the excitotoxic effects of glutamate. Pretreatment of cerebellar neurons with the AMPA antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione blocked the neuroprotective effect mediated by AMPA indicating that the neuroprotective effect is mediated specifically by AMPA receptors. An excitotoxic concentration of glutamate, which killed between 60-80% of granule cell neurons on day 8 in vitro, mediated its toxic effect via a time-dependent apoptotic pathway. Pretreatment of cerebellar granule cell neurons with AMPA (500 microM) completely blocked glutamate-mediated apoptosis. Our results suggest that AMPA receptors may play an important role in neuronal survival. PMID- 15545007 TI - Protection by conventional and new antiepileptic drugs against lindane-induced seizures and lethal effects in mice. AB - Toxic effects caused by the organochlorine xenobiotic lindane may result from too excessive antiscabicidal treatment and in cases of accidental or intentional poisoning. Predominant symptoms of lindane intoxication concern the central nervous system, e.g. different manifestation of hyperexcitability and epileptiform activity. The inhibition of GABA-ergic neurotransmission seems to be responsible for the convulsant activity of lindane. This study was intended to compare the protective activity of conventional and new antiepileptic drugs against convulsions and lethal effects elicited by lindane administration in mice. Diazepam, clonazepam and phenobarbital protected against full seizure pattern and lethal effects evoked by lindane. Carbamazepine, phenytoin, gabapentin, felbamate and lamotrigine inhibited only lindane-induced tonic convulsions and mortality. It may be concluded that apart from benzodiazepines, phenobarbital and, to a lesser extent, carbamazepine, phenytoin, gabapentin, felbamate and lamotrigine could be used in lindane poisoning. Vigabatrin proved completely ineffective against seizures or lethal effects elicited by lindane. PMID- 15545008 TI - Flotillin-1 in the substantia nigra of the Parkinson brain and a predominant localization in catecholaminergic nerves in the rat brain. AB - The substantia nigra cells of a normal and Parkinson's disease human brain were obtained by the micropunch procedure and total RNA was isolated. Differential display RT-PCR of the total RNA revealed differentially expressed cDNAs that were identified by sequencing. This resulted in the identification of a panel of known and unknown differentially expressed genes. Complex I (NADH ubiquinone oxidoreductase) and Complex IV (cytochrome oxidase) whose expressions are decreased in Parkinson's disease were reduced in the Parkinson brain. Of the various differentially expressed genes, flotillin-1, also known as reggie-2, was of great interest to us. It is a relatively new protein which is an integral membrane component of lipid rafts and has been implicated in signal transduction pathway events. In situ hybridization histochemical studies with human and rat brain sections revealed the presence of this mRNA in discrete neuronal (and possibly glial) cells of the substantia nigra, locus coeruleus, cortex, hippocampus, hypothalamus, thalamus, motor nuclei, nucleus basalis, raphe nucleus, and other brain regions. Immunohistochemical studies revealed that flotillin-1 is not present in all the regions where the message was found. In the rat brain, the most prominent observation was the revelation of all catecholamine cells (dopamine, norepinephrine, epinephrine) by the flotillin-1 antibody (1:100 dilution). At a more concentrated dilution (1:10) other neuronal cells (e.g., cortex, thalamus, hindbrain) were observed. At both dilutions dense dopaminergic fibers were observed in the rat caudate-putamen, nigrostriatal tract, and substantia nigra. It is significant that there is an increased gene expression of flotillin-1 in the Parkinson substantia nigra/ventral tegmental area. The role of flotillin in these cells is unclear although it is interesting that the reggie 2/flotillin-1 gene was upregulated during retinal axon regeneration in the goldfish visual pathway (Schulte et al., Development 124:577-87, 1997) which suggests that flotillin-1/reggie-2 might play a role in axonal growth from the remaining substantia nigra cells of the Parkinson brain. PMID- 15545009 TI - Central depletion of angiotensinogen is associated with elevated AT1 receptors in the SFO and PVN. AB - The brain renin-angiotensin system (RAS) is important in fluid balance and blood pressure regulation. In this study, we compared angiotensin (Ang) receptor density in the subfornical organ (SFO) and paraventricular nucleus (PVN) of a) brain angiotensinogen deficient rats (ASrAogen); b) those with high levels of brain Ang II [(mRen2)27]; c) Hannover Sprague Dawley (SD) rats at 48 and 68 wks of age. Since there was no difference between the two ages in any of the three strains, the data from the 48 and 68 wk time points were combined. There was a significantly higher level of AT1 receptors in the SFO and PVN of ASrAogen animals compared to both the SD and (mRen2)27 rats. This suggests that the brain RAS is important in regulating receptor density and that the differences may be explained by lower levels of the peptide locally. These higher levels of receptors suggest that the ASrAogen animals in adulthood and early aging would be more sensitive to either circulating or endogenous brain Ang II than the SD animals of similar age. In contrast, the similar receptor density in the (mRen2)27 and SD rats suggest that previous reports of reduced responses in the (mRen2)27 rats may result from differences in post receptor mechanisms such as intracellular signaling. Moreover, our data reveal that functional assessments are necessary in addition to receptor density levels to understand the consequences of long-term alterations in brain tissue peptides. PMID- 15545010 TI - Postnatal apoptosis in cerebellar granule cells of homozygous leaner (tg1a/tg1a) mice. AB - Leaner mice carry a homozygous, autosomal recessive mutation in the mouse CACNA1A gene encoding the Alpha1A subunit of P/Q-type calcium channels, which results in an out-of-frame splicing event in the carboxy terminus of the Alpha1A protein. Leaner mice exhibit severe ataxia, paroxysmal dyskinesia and absence seizures. Functional studies have revealed a marked decrease in calcium currents through leaner P/Q-type channels and altered neuronal calcium ion homeostasis in cerebellar Purkinje cells. Histopathological studies of leaner mice have revealed extensive postnatal cerebellar Purkinje and granule cell loss. We examined the temporospatial pattern of cerebellar granule cell death in the leaner mouse between postnatal days (P) 10 and 40. Our observations clearly indicate that leaner cerebellar granule cells die via an apoptotic process and that the peak time of neuronal death is P20. We did not observe a significant increase in microglial and astrocytic responses at P20, suggesting that glial responses are not a cause of neuronal cell death. We propose that the leaner cerebellar granule cell represents an in vivo animal model for low intracellular [Ca2+]-induced apoptosis. Since intracellular [Ca2+] is critical in the control of gene expression, it is quite likely that reduced intracellular [Ca2+] could activate a lethal cascade of altered gene expression leading to the apoptotic granule cell death in the leaner cerebellum. PMID- 15545011 TI - Inhibition of protein kinase C promotes neuronal survival in low potassium through an Akt-dependent pathway. AB - Cerebellar granule cell neurons undergo apoptotic cell death when subjected to serum-free conditions at physiological concentrations of potassium (5 mM). Protein kinase C (PKC) is known to play a role in preventing neuronal apoptosis under trophic factor deprivation, but its role in protecting cerebellar neurons from cell death under conditions of low potassium is unknown. This study sought to determine the involvement of PKC in neuronal survival and to determine if PKC regulated the phosphatidylinositol 3-kinase (PI 3-K)/Akt pathway in low physiologic concentrations of potassium. Incubation with a pan-PKC inhibitor, Ro 31-8220 (2 microm), or a specific PKCAlpha inhibitor, Go6976, protected cerebellar granule cell neurons from low potassium-mediated cell death. In contrast, phorbol ester (TPA, 100 nm), a PKC activator, increased cell death. Incubation with, Ro-31-8220 rescued neurons from cell death induced by the PI 3-K inhibitor, LY294002, suggesting that Ro-31-8220 may affect Akt phosphorylation. Western blot analysis showed that serum-free, low potassium conditions decreased Akt phosphorylation, which was exacerbated by treatment with LY294002. In contrast, PKC inhibitors, Go6976 or Ro-31-8220, increased Akt phosphorylation approximately two and four-fold, respectively in low potassium conditions. Because Akt activation appears to be critical in promoting neuronal survival under these culture conditions, increased Akt phosphorylation brought about by inhibiting PKC promotes neuronal survival. PMID- 15545012 TI - Purinergic-receptor oligomerization: implications for neural functions in the central nervous system. AB - It is becoming clear that the functions of G protein-coupled receptors (GPCRs), the largest family of plasma membrane-localized receptors, are regulated by direct oligomeric formation between GPCRs, as either homo- or hetero-oligomers. This review article explores the mechanistic implications of GPCR dimerization, especially among purinergic receptors, adenosine receptors and P2 receptors, which play critical roles in the regulation of neurotransmission in the central nervous system. Briefly, adenosine receptors are able to form a heteromeric complex with P2 receptors that generates an adenosine receptor with P2 receptor like agonistic pharmacology. This mechanism may be used to fine-tune purinergic inhibition locally at sites where there is a particular oligomerization structure between purinergic receptors, and to explain the undefined adenosine-like purinergic functions of adenine nucleotides. Purinergic receptors also form oligomers with GPCRs of other families present in the brain, such as dopamine receptors and metabotropic glutamate receptors, to alter the functional properties. The effect of GPCR oligomerization on receptor functions is thus considered as an important system in the central nervous system. PMID- 15545013 TI - Ovariectomy combined with amyloid beta(1-42) impairs memory by decreasing acetylcholine release and alpha 7nAChR expression without induction of apoptosis in the hippocampus CA1 neurons of rats. AB - In this study, the effect of ovariectomy and amyloid Beta(1-42) (ABeta(1-42))on eight-armed radial maze performance, acetylcholine (ACh) release, Alpha7nACh receptor (Alpha7nAChr), glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression, and apoptosis of CA1 neurons in the dorsal hippocampus were investigated in rat. The results showed that the dorsal hippocampus of sham rats contains 136.7 -/+ 16.7 to 160.4 -/+ 21.1 fmol/microl ACh, and respective 201 -/+ 22.9 and 416.6 -/+ 66.3 expression of mRNA for a7nAChR and GAPDH. Ovariectomy alone, after 4 weeks, did not impair memory, and neither induced apoptosis nor changed the basal ACh release. On the other hand, ABeta(1-42) (600 pmol/10 microl/body/day i.c.v. for 7 days) impaired memory, an effect characterized by increased error choices and reduced (50-59%) ACh release, but only with slight apoptosis. Moreover, ovariectomy combined with ABeta(1-42) induced memory impairment characterized by decreased numbers of correct choices and increased numbers of errors. This effect was accompanied by a decrease of the basal ACh level (67%), a7nAChR mRNA expression (52%) and a7nAChR/GAPDH ratio (44%) without induction of apoptosis in the dorsal hippocampus. The high K+-evoked ACh release was not altered in ovariectomized rats, but was decreased by ABeta(1-42) (43%) and ovariectomy + ABeta(1-42) (80%). These results suggest that ovariectomy induced hormonal deprivation after 4 weeks, when accompanied by ABeta(1-42) accumulation in the dorsal hippocampus, could impair memory by decreasing ACh release and a7nAChR expression without inducing apoptosis in the CA1 field of the dorsal hippocampus. PMID- 15545014 TI - Nicotine inhibits ethanol-induced toxicity in cultured cerebral cortical cells. AB - The high incidence of smoking among alcoholics may be partially due to nicotine's ability to counteract some of the adverse effects of ethanol on motor coordination and/or cognitive functions. Neuroprotective effects of nicotine on ethanol-induced toxicity in cerebellar granular cells have been observed. In this study, we sought to determine whether similar protection is observed in neocortical cells and if so, what specific nicotinic receptor subtypes may be mediating the actions of nicotine. Primary cultures of neocortical cells were prepared from 20-day embryos obtained from time-pregnant Sprague-Dawley rats. Cells were cultured for 10 days and were then exposed for 3 days to various concentrations of ethanol with and without pretreatment with nicotine and nicotinic antagonists. Cellular toxicity was evaluated by measuring the lactate dehydrogenase level. Administration of ethanol (10-100 mM) resulted in a dose dependent toxicity. Pretreatment with nicotine 5-20 microM resulted in a dose dependent protection against ethanol-induced toxicity. The effects of nicotine were blocked by pretreatment with nicotinic antagonists such as mecamylamine (1 20 microM), dihydro-beta-erythroidine (DHBE) 50 nM-1.0 microM and methyllycaconitine (MLA) 5 nM-1 microM in a dose-dependent manner. Compared to previous studies, higher ethanol concentrations were required to induce toxicity in neocortical vs cerebellar granule cells. Moreover, the effects of nicotine in the neocortical cells were blocked by lower concentrations of MLA, but higher concentrations of DHBE compared to cerebellar cells. Collectively, the results suggest differential sensitivity of various neuronal populations to the toxic effect of ethanol. Furthermore, protective effects of nicotine against alcohol in various regions appear to be mediated by different nicotinic receptor subtypes. The neuroprotective effect of nicotine against ethanol-induced toxicity may be a contributing factor to the high incidence of smoking among alcoholics. PMID- 15545015 TI - Serotoninergics attenuate hyperlocomotor activity in rats. Potential new therapeutic strategy for hyperactivity. AB - Hyperactivity is thought to be associated with an alteration of dopamine (DA) neurochemistry in brain. This conventional view became solidified on the basis of observed hyperactivity in DA-lesioned animals and effectiveness of the dopaminomimetics such amphetamine (AMP) in abating hyperactivity in humans and in animal models of hyperactivity. However, because AMP releases serotonin (5-HT) as well as DA, we investigated the potential role of 5-HT in an animal model of hyperactivity. We found that a greater intensity of hyperactivity was produced in rats when both DA and 5-HT neurons were damaged at appropriate times in ontogeny. Therefore, previously we proposed this as an animal model of attention deficit hyperactivity disorder (ADHD) - induced by destruction of dopaminergic neurons with 6-hydroxydopamine (6-OHDA) (neonatally) and serotoninergic neurons with 5,7 dihydroxytryptamine (5,7-DHT) (in adulthood). In this model effects similar to that of AMP (attenuation of hyperlocomotion) were produced by m chlorophenylpiperazine (m-CPP) but not by 1-phenylbiguanide (1-PG), respective 5 HT2 and 5-HT3 agonists. The effect of m-CPP was shown to be replicated by desipramine, and was largely attenuated by the 5-HT2 antagonist mianserin. These findings implicate 5-HT neurochemistry as potentially important therapeutic targets for treating human hyperactivity and possibly childhood ADHD. PMID- 15545016 TI - Studies on homocysteine and dehydroepiandrosterone sulphate plasma levels in Alzheimer's disease patients and in Parkinson's disease patients. AB - Homocysteine (HC) and dehydroepiandrosterone sulphate (DHEAS) plasma levels have been evaluated in groups of male and female patients with Parkinson's disease (PD) and in a group of female patients with Alzheimer's disease (AD) and compared with the corresponding plasma levels observed in a group of age-matched subjects. It has been confirmed that HC plasma levels are enhanced in both PD and AD patients. As far as the DHEAS plasma levels are concerned no changes have been observed in PD patients while a marked decrease has been observed in AD patients. These results support the view that while the pro-oxidant effects of HC and its agonist action at NMDA receptors can play a role in both neurodegenerative diseases, the role of DHEAS is more complex and may be an important factor only in certain neurodegenerative diseases. Thus, according to the present study DHEAS is likely to be involved in AD but not in PD. PMID- 15545017 TI - Common patterns of bcl-2 family gene expression in two traumatic brain injury models. AB - Cell death/survival following traumatic brain injury (TBI) may be a result of alterations in the intracellular ratio of death and survival factors. Bcl-2 family genes mediate both cell survival and the initiation of cell death. Using lysate RNase protection assays, mRNA expression of the anti-cell death genes Bcl 2 and Bcl-xL, and the pro-cell death gene Bax, was evaluated following experimental brain injuries in adult male Sprague-Dawley rats. Both the lateral fluid-percussion (LFP) and the lateral controlled cortical impact (LCI) models of TBI showed similar patterns of gene expression. Anti-cell death bcl-2 and bcl-xL mRNAs were attenuated early and tended to remain depressed for at least 3 days after injury in the cortex and hippocampus ipsilateral to injury. Pro-cell death bax mRNA was elevated in these areas, usually following the decrease in anti-cell death genes. These common patterns of gene expression suggest an important role for Bcl-2 genes in cell death and survival in the injured brain. Understanding the regulation of these genes may facilitate the development of new therapeutic strategies for a condition that currently has no proven pharmacologic treatments. PMID- 15545019 TI - Functional genomics strategies to identify susceptibility genes and treatment targets in alcohol dependence. AB - Genetic factors contribute to alcohol dependence through two main categories of mechanisms. The 50-60% heritability observed in this disorder is presumably conferred by polymorphic variants, encoding functionally altered proteins, or leading to differential transcriptional activity. Secondly, long term changes during the process of developing dependence are likely encoded by persistent changes in gene expression. Thus, genetic and environmental factors interact at the level of the transcriptome, making this an attractive level of analysis. For this purpose, we have applied differential display and more recently Affymetrix oligonucleotide gene arrays to models of genetic susceptibility and alcohol induced neuroadaptation. PMID- 15545020 TI - IDRD2 TaqIA polymorphism is associated with urinary homovanillic acid levels in a sample of Spanish male alcoholic patients. AB - The TaqIA1 allele of the dopamine receptor gene D2 (DRD2) has been associated with alcoholism, as well as with other addictive behaviours. The exact nature of how the presence of this allele can be a vulnerability factor in the development of alcoholism remains unclear. In this study we found that the presence in the DRD2 genotype of the TaqIA1 allele in Spanish alcoholics is associated with higher levels of urine homovanillic acid (HVA) when compared to patients homozygous for the TaqIA2 allele. A sample of 142 Spanish male alcoholic patients was split into 2 groups on the basis of the presence or absence of the A1 allele in their genotype. The urine sample was analyzed by high performance liquid cromatography (HPLC), and the concentration of homovanillic acid (HVA), 5 hydroxyindoleacetic acid (5-HIAA) and vanilylmandelic acid (VMA) was determined. We found a statistical difference in the concentration of HVA between the groups, that suggests this polymorphism could be related to the variance of urine HVA levels. PMID- 15545021 TI - The role of the dopamine transporter in cocaine abuse. AB - There have been many studies aimed at understanding the role that the dopamine transporter plays in cocaine abuse. Most studies suggest that inhibition of dopamine uptake by cocaine is the primary mechanism by which its behavioral effects are produced. Because of the strong relationship between binding to the dopamine transporter and the behavioral effects of cocaine, the dopamine transporter has on occasion been referred to as the cocaine binding site. Chronic studies using cocaine or selective inhibitors of dopamine, norepinephrine, or serotonin uptake suggest that while a selective dopamine uptake inhibitor can produce sensitization to cocaine, the long-lasting sensitized response to a cocaine challenge observed in cocaine-pretreated rats is due to cocaine's action on a system other than, or in addition to, dopamine. Thus, while dopamine appears to be important for the behavioral effects of cocaine, it appears that neurochemical systems other than dopamine likely play a role in the behavioral effects of chronic cocaine. PMID- 15545022 TI - Homer2 gene deletion in mice produces a phenotype similar to chronic cocaine treated rats. AB - Addiction to cocaine results from changes in brain function arising from a combination of pharmacology, environmental circumstances, as well as genetic vulnerability. One change is a reduction in Homer protein in the nucleus accumbens. In this report we summarize the behavioral and neurochemical effects of Homer2 gene deletion in mice and compare this with the effects of chronic cocaine treatment in rats. It was shown that Homer2 KO mice demonstrate enhanced locomotor stimulant and conditioned place preference responses to cocaine. Homer2 deletion also caused mice to show reduced basal extracellular glutamate in the nucleus accumbens and a sensitized increase in extracellular glutamate in response to a cocaine injection. In contrast to glutamate, Homer2 KO mice showed a normal increase in extracellular dopamine following a cocaine challenge injection. The parallel between the effect of Homer2 gene deletion and chronic cocaine administration on behavioral and glutamatergic neurochemical responses to cocaine supports involvement of Homer proteins and glutamate transmission in the sensitization of behavior produced by repeated cocaine. PMID- 15545018 TI - Gene-environment interplay in alcoholism and other substance abuse disorders: expressions of heritability and factors influencing vulnerability. AB - Factors that confer predisposition and vulnerability for alcoholism and other substance abuse disorders may be described usefully within the gene-environment interplay framework. Thus, it is postulated that heritability provides a major contribution not only to alcohol but also to other substances of abuse. Studies of evoked potential amplitude reduction have provided a highly suitable and testable method for the assessment of both environmentally-determined and heritable characteristics pertaining to substance use and dependence. The different personal attributes that may co-exist with parental influence or exist in a shared, monozygotic relationship contribute to the final expression of addiction. In this connection, it appears that personality disorders are highly prevalent co-morbid conditions among addicted individuals, and, this co-morbidity is likely to be accounted for by multiple complex etiological relationships, not least in adolescent individuals. Co-morbidity associated with deficient executive functioning may be observed too in alcohol-related aggressiveness and crimes of violence. The successful intervention into alcohol dependence and craving brought about by baclofen in both human and animal studies elucidates glutamatergic mechanisms in alcoholism whereas the role of the dopamine transporter, in conjunction with both the noradrenergic and serotonergic transporters, are implicated in cocaine dependence and craving. The role of the cannabinoids in ontogeny through an influence upon the expression of key genes for the development of neurotransmitter systems must be considered. Finally, the particular form of behaviour/characteristic outcome due to childhood circumstance may lie with biological, gene-based determinants, for example individual characteristics of monoamine oxidase (MAO) activity levels, thereby rendering simple predictive measures both redundant and misguiding. PMID- 15545023 TI - Cannabinoids and gene expression during brain development. AB - Cannabis is the most commonly used illicit drug in western societies, in particular among young people. It is consumed even by women during pregnancy and lactation, which result in a variety of disturbances in the development of their offspring, because, like other habit-forming drugs, cannabinoids, the psychoactive ingredients of marijuana, can cross the placental barrier and be secreted in the maternal milk. Through this way, cannabinoids affect the ontogeny of various neurotransmitter systems leading to changes in different behavioral patterns. Dopamine and endogenous opioids are among the neurotransmitters that result more affected by perinatal cannabinoid exposure, which, when animals mature, produce changes in motor activity, drug-seeking behavior, nociception and other processes. These disturbances are likely originated by the capability of cannabinoids to influence the expression of key genes for both neurotransmitters, in particular, the enzyme tyrosine hydroxylase and the opioid precursor proenkephalin. In addition, cannabinoids seem to be also able to influence the expression of genes encoding for neuron-glia cell adhesion molecules, which supports a potential influence of cannabinoids on the processes of cell proliferation, neuronal migration or axonal elongation in which these proteins are involved. In support of this possibility, CB1 receptors, which represent the major targets for the action of cannabinoids, are abundantly expressed in certain brain regions, such as the subventricular areas, which have been involved in these processes during brain development. Finally, cannabinoids might also be involved in the apoptotic death that occurs during brain development, possibly by influencing the expression of Bcl-2/Bax system. Also in support of this option, CB1 receptors are transiently expressed during brain development in different group of neurons which do not contain these receptors in the adult brain. This paper will review all evidence relating cannabinoids to the expression of key genes for neural development, trying to establish the future research addressed to elucidate the mechanisms involved in the epigenetic action of cannabinoids during brain development. PMID- 15545025 TI - Duration of immunity produced by a live attenuated vaccine against avian pneumovirus type C. AB - A recently developed live, attenuated vaccine against avian pneumovirus (APV) was found to be safe and protective in experimental birds. Duration of immunity following a single dose of this experimental vaccine in 1-week-old turkey poults is described. Two groups each of 60 poults were housed in separate isolation rooms. Birds in group one were inoculated oculonasally at 1 week of age with the vaccine. The second group served as a non-vaccinated group and was inoculated with mock-infected cell culture fluid. At 3, 7, 10, and 14 weeks post vaccination, 15 birds from each of the groups were removed to separate isolation rooms and challenged with virulent APV. Taken together, data on clinical signs and virus detection in choanal swabs following each challenge indicated that the vaccine was able to protect birds for up to 14 weeks post vaccination. Peak antibody levels were attained 7 weeks post vaccination and declined thereafter. These results indicated that this experimental vaccine induced protection against APV even in the absence of high antibody titres. PMID- 15545024 TI - Role of GABA(B) receptor in alcohol dependence: reducing effect of baclofen on alcohol intake and alcohol motivational properties in rats and amelioration of alcohol withdrawal syndrome and alcohol craving in human alcoholics. AB - The present paper describes the results of recent preclinical and clinical studies conducted in this laboratory in order to characterize the anti-alcohol properties of the GABA(B) receptor agonist, baclofen. At a preclinical level, the repeated administration of non-sedative doses of baclofen dose-dependently suppressed the acquisition and maintenance of alcohol drinking behavior in selectively bred Sardinian alcohol-preferring (sP) rats tested under the homecage, 2-bottle "alcohol vs water" choice regimen. Acute injection of baclofen completely blocked the temporary increase in voluntary alcohol intake occurring after a period of alcohol abstinence (the so-called alcohol deprivation effect, which models alcohol relapses in human alcoholics). Acute treatment with baclofen also dose-dependently suppressed extinction responding for alcohol (an index of motivation to consume alcohol) in sP rats trained to lever-press for oral alcohol self-administration. Taken together, these results suggest the involvement of the GABA(B) receptor in the neural substrate mediating alcohol intake and alcohol motivational properties in an animal model of excessive alcohol consumption. Further, acutely administered baclofen dose-dependently reduced the severity of alcohol withdrawal signs in Wistar rats made physically dependent upon alcohol. Preliminary clinical surveys suggest that the anti-alcohol properties of baclofen observed in rats may generalize to human alcoholics. Indeed, a double-blind survey demonstrated that repeated daily treatment with baclofen was associated, when compared to placebo, with a higher percentage of subjects totally abstinent from alcohol and a higher number of days of total abstinence. Treatment with baclofen also suppressed the number of daily drinks and decreased the obsessive and compulsive components of alcohol craving. Finally, a single non-sedative dose of baclofen resulted in the rapid disappearance of alcohol withdrawal symptomatology, including delirium tremens, in alcohol-dependent patients. In both clinical studies, baclofen was well tolerated with minimal side effects. These results suggest that baclofen may represent a potentially effective medication in the treatment of alcohol-dependent patients. PMID- 15545026 TI - Assessment of genetic, antigenic and pathotypic criteria for the characterization of IBDV strains. AB - The aim of this work was the selection and comparison of representative infectious bursal disease virus (IBDV) strains. Nine strains of IBDV, isolated at different times and from different geographic regions of Europe and China, were characterized. Batches of all strains were prepared following standardized protocols and checked for the absence of contaminating viruses. Criteria used for their characterization were: (i) the nucleotide sequence of the VP2 variable region, (ii) binding to a panel of neutralizing monoclonal antibodies in antigen capture enzyme-linked immunosorbent assays, and (iii) virulence in specific pathogen free chickens after infection with a standardized number of median embryo infective doses. Based on the first two criteria, two of nine strains were classified as classical virulent (cv) IBDV (F52/70, Cu-1wt), and five as very virulent (vv) IBDV (849VB, 96108, HK46, GX, Harbin). Remarkably, although a clear cut difference was demonstrable between European cvIBDV (F52/70 and Cu-1wt) and vvIBDV (849VB and 96108) strains, there was a continuum in the pathogenicity of Chinese vvIBDVs. Our results indicate the probable existence of differences in virulence within IBDV lineages determined on the basis of antigenic typing using monoclonal antibodies and the alignment of the VP2 sequences. This indicates limitations in the analysis of IBDV pathotypes based on the VP2 variable region and emphasizes that these criteria may not be sufficient for the classification of IBDV strains. PMID- 15545027 TI - Comparitive sensitivity of polymerase chain reaction diagnosis of psittacine beak and feather disease on feather samples, cloacal swabs and blood from budgerigars (Melopsittacus undulates, Shaw 18005). AB - A longitudinal study was performed in order to investigate virus excretion and viraemia during a clinical outbreak of the psittacine beak and feather disease in budgerigars (Melopsittacus undulatus). Viral nucleic acid was detected in feathers, cloacal swabs and blood samples. Overall, beak and feather disease virus (BFDV) DNA was detected most commonly in feather samples, followed by cloacal swabs, and least frequently from blood samples. In most cases the viraemia was short lived and correlated with clinical signs, such as feather abnormalities. Sequence analysis of the polymerase chain reaction fragment amplified from the replication-associated gene (ORF V1) indicated a close relationship with other BFDV isolates. Overall the highest level of nucleotide identity was found with the ORF V1 of another budgerigar isolate. Our results suggest that feather samples and cloacal swabs should be taken for polymerase chain reaction diagnosis to determine the presence of BFDV in an aviary, but that detection in these samples may not correlate well with psittacine beak and feather disease. PMID- 15545028 TI - Molecular epidemiology of Newcastle disease viruses isolated in South Korea using sequencing of the fusion protein cleavage site region and phylogenetic relationships. AB - The study, using sequence analysis and the phylogenetic relationship of the fusion protein gene, divided the Korean epizootic isolates of Newcastle disease virus (NDV) into several lineages to determine the molecular epidemiology of the virus. A 695 base pair fragment was amplified by polymerase chain reaction between matrix protein gene and fusion protein gene of 30 Korean NDV isolates, which were isolated from field outbreaks of Newcastle disease between 1949 and 2002. All isolates showed the amino acid sequence 112R-R-Q/R-K-R116 at the C terminus of the F2 protein and phenylalanine (F) at the N-terminus of the F1 protein, residue 117. These amino acid sequences were identical to a known virulent motif. The region of the F gene between nucleotides 47 and 435 was compared by phylogenetic analysis. Based on nucleotide sequence, the Korean NDV isolates belonged to genotype III, V, VI and VII corresponding to isolates in 1949, 1982 to 1984, 1988 to 1997, and 1995 to 2002, respectively. These data showed that genotypes of five Korean Newcastle disease epizootics had replaced each other serially (III, V, VI and VII) in chronological order. Further, the five Korean Newcastle disease epizootics were closely related with the Newcastle disease panzootics or Newcastle disease epizootics in other countries. Present study showed that the Korean genotype V isolated before 1984 was related with European Newcastle disease epizootics in the 1970s, whereas the Korean genotypes VI and VII isolated after 1988 were more closely related with Far East Newcastle disease epizootics, especially Newcastle disease epizootics in Japan, Taiwan and China. Since 1988, the genotypes VI and VII of Far East origin were dominant in South Korea. That might be due to the increased trade of agricultural products including poultry among Far East Asian countries. PMID- 15545029 TI - Investigation of outbreaks of highly pathogenic H5N1 avian influenza in waterfowl and wild birds in Hong Kong in late 2002. AB - Outbreaks of highly pathogenic H5N1 avian influenza have occurred in Hong Kong in chickens and other gallinaceous poultry in 1997, 2001, twice in 2002 and 2003. High mortality rates were seen in gallinaceous birds but not in domestic or wild waterfowl or other wild birds until late 2002 when highly pathogenic H5N1 avian influenza occurred in waterfowl (geese, ducks and swans), captive Greater Flamingo (Phoenicopterus ruber) and other wild birds (Little Egret Egretta garzetta) at two waterfowl parks and from two dead wild Grey Heron (Ardea cinerea) and a Black-headed Gull (Larus ridibundus) in Hong Kong. H5N1 avian influenza virus was also isolated from a dead feral pigeon (Columba livia) and a dead tree sparrow (Passer montanus) during the second outbreak. The first waterfowl outbreak was controlled by immediate strict quarantine and depopulation 1 week before the second outbreak commenced. Control measures implemented for the second outbreak included strict isolation, culling, increased sanitation and vaccination. Outbreaks in gallinaceous birds occurred in some live poultry markets concurrently with the second waterfowl outbreak, and infection on a chicken farm was detected 1 week after the second waterfowl park outbreak was detected, on the same day the second grey heron case was detected. Subsequent virus surveillance showed the outbreaks had been contained. PMID- 15545030 TI - Cross-protection study of the nine serovars of Haemophilus paragallinarum in the Kume haemagglutinin scheme. AB - The cross-protection and haemagglutination-inhibition antibodies present in chickens vaccinated with one of the nine currently recognized Kume haemagglutinin serovars of Haemophilus paragallinarum were investigated. The results confirmed the widely accepted dogma that serogroups A, B, and C represent three distinct immunovars. Within Kume serogroup A, there was generally good cross-protection among all four serovars. However, within Kume serogroup C, there was evidence of a reduced level of cross-protection between some of the four serovars. The haemagglutination-inhibition antibody levels generally showed the same trend as with the cross-protection results. This study suggests that some apparent field failures of infectious coryza vaccines may be due to a lack of cross-protection between the vaccine strains and the field strains. Our results will help guide the selection of strains for inclusion in infectious coryza vaccines. PMID- 15545031 TI - Heat inactivation of avian influenza and Newcastle disease viruses in egg products. AB - Avian influenza (AI) and Newcastle disease (ND) viruses are heat labile viruses, but exact parameters for heat inactivation at egg pasteurization temperatures have not been established. In this study we artificially infected four egg products with two AI (one low [LP] and one high pathogenicity [HP]) and three ND (two low and one highly virulent) viruses, and determined inactivation curves at 55, 57, 59, 61 and 63 degrees C. Based on D(t) values, the time to inactivation of the viruses was dependent on virus strain and egg product, and was directly related to virus titre, but inversely related to temperature. For all temperatures, the five viruses had the most rapid and complete inactivation in 10% salt yolk, while the most resistant to inactivation was HPAI virus in dried egg white. This study demonstrated that the LPAI and all ND viruses were inactivated in all egg products when treated using industry standard pasteurization protocols. By contrast, the HPAI virus was inactivated in liquid egg products but not in dried egg whites when using the low-temperature industry pasteurization protocol. PMID- 15545032 TI - Pathology of lymphoid organs in chickens fed a diet deficient in zinc. AB - An experiment was conducted, including flow cytometry, to study the pathology of the lymphoid organs and peripheral blood T lymphocytes in zinc (Zn)-deficient chickens. One hundred 1-day-old broiler chickens were randomly divided into two groups and fed on diets with 100 mg/kg Zn (controls) or Zn-deficient diets (Zn, 23.63 mg/kg) for 7 weeks. The weight and growth index of the bursa of Fabricius, thymus and spleen were significantly reduced (P<0.05 or P<0.01) in Zn-deficient birds when compared with those of control broilers. The G0/G1 phase of the cell cycle of the bursa, thymus and spleen was much higher (P<0.01), and the S, G2+M phases and proliferating index lower (P<0.05 or P<0.01) in Zn-deficient broilers than in the controls. The acid alpha-naphthyl acetate esterase-positive ratio of the peripheral blood T lymphocytes and the CD4 and CD8 numbers were markedly reduced (P<0.05 or P<0.01), and the CD4/CD8 ratio increased. Histopathologically, lymphocytes of lymphoid organs were depleted and the reticular cells of the thymus were also degenerate or necrotic in the Zn-deficient birds. The results demonstrate that Zn deficiency seriously inhibited the development of lymphoid organs, impaired the progression of lymphocytes from the G0/G1 phase to the S phase, and caused pathological injury in the lymphoid organs. The results also showed that the effect of Zn deficiency on the primary lymphoid organs occurred earlier than on the secondary lymphoid organs. The effect of Zn deficiency was greatest on the bursa of Fabricius, followed by the thymus, and then the spleen. Potential mechanisms underlying the observations are discussed. PMID- 15545033 TI - Circovirus infection in a Gouldian finch (Chloebia gouldiae). AB - The bird examined was a 10-week-old female Gouldian finch (Chloebia gouldiae) from an aviary that had housed about 100 Gouldian finches, which had nasal discharge, dyspnoea, anorexia, depression and a very high mortality (50%) in both adult and young birds. Gross and histopathology revealed moderate to severe lymphoid depletion in the bursa of Fabricius and thymus, and sinusitis/rhinitis, tracheitis, bronchopneumonia, myocarditis, nephritis and splenitis. Circovirus infection was diagnosed in the Gouldian finch based on finding characteristic globular intracytoplasmic inclusion bodies containing 15 to 18 nm virus particles in the mononuclear cells of the bursa of Fabricius by transmission electron microscopy and by demonstrating circovirus DNA in the cytoplasm of mononuclear cells of the bursa of Fabricius by in situ hybridization using a circovirus specific DNA probe. The Gouldian finch was also affected by concurrent bacterial and adenovirus infections. This is the first report of circovirus infection in a Gouldian finch. PMID- 15545035 TI - Detection of Cryptosporidium oocysts and Giardia cysts in swimming pool filter backwash water concentrates by flocculation and immunomagnetic separation. AB - The purpose of the present study was to evaluate techniques for detection of Giardia cysts and Cryptosporidium oocysts in swimming pool filter backwash water. Calcium carbonate flocculation was used for water samples concentration of 1 l filter backwash water samples. Immunomagnetic separation (IMS) was used for separation of cysts and oocysts from detrital material. Cysts and oocysts were then detected using direct immunofluorescence. ColorSeed C&G was used as an internal standard. Recoveries were examined at several processing points. Highest recoveries (23% Cryptosporidium, 18% Giardia) were obtained using lower filter backwash volumes, greater IMS volumes, and addition of Tween20 to backwash samples prior to processing. The combination of CaCO3 flocculation, IMS and direct immunofluorescence was found to be an effective tool for the detection and quantification of Giardia spp. and Cryptosporidium spp. in filter backwash water samples. PMID- 15545036 TI - Qat chewing and pesticides: a study of adverse health effects in people of the mountainous areas of Yemen. AB - Chewing qat leaves, Cathula Edulis, is now a very common behaviour among the people of the mountainous areas of Yemen. For about the past 20 years, in tandem with national development, qat chewing has been rapidly expanding, and the use of chemical pesticides in qat production has been increasing. In this study, the adverse effects on human health of qat chewing combined with pesticide use were investigated. Results of interviews and questionnaires showed that chewers of qat grown with few or no chemical pesticides and chewers of qat grown with chemical pesticides have considerably different subjective symptoms. Chewers of qat produced in fields where chemical pesticides are used regularly have more symptoms than chewers of qat produced in fields where chemical pesticides are rarely or never used. Chewers of qat produced with more chemical pesticides, in particular, experience acute adverse effects on the digestive system and chronic adverse effects such as body weakness and nasal problems. Farmers who chew homemade qat on which they spread chemical pesticides by themselves may have the highest health risks regarding the combination of qat and pesticides. It is concluded that chewing qat grown with chemical pesticides causes considerable adverse health effects in human beings. PMID- 15545038 TI - Climatological variables and the incidence of Dengue fever in Barbados. AB - A retrospective study to determine relationships between the incidence of dengue cases and climatological variables and to obtain a predictive equation was carried out for the relatively small Caribbean island of Barbados which is divided into 11 parishes. The study used the weekly dengue cases and precipitation data for the years (1995 - 2000) that occurred in the small area of a single parish. Other climatological data were obtained from the local meteorological offices. The study used primarily cross correlation analysis and found the strongest correlation with the vapour pressure at a lag of 6 weeks. A weaker correlation occurred at a lag of 7 weeks for the precipitation. The minimum temperature had its strongest correlation at a lag of 12 weeks and the maximum temperature a lag of 16 weeks. There was a negative correlation with the wind speed at a lag of 3 weeks. The predictive models showed a maximum explained variance of 35%. PMID- 15545037 TI - Effects of mould remediation on school teachers' health. AB - The follow-up study of the health of teachers (n = 56) of three mould damage schools were done with self-administered symptom questionnaire before and 1 year after the remediation of school buildings. Technical and microbiological investigations were done parallel at the same time. In the beginning of the study symptoms of allergic rhinitis, sinusitis, conjunctivitis and fatigue were high compared to normal population and 1 year after the intervention a decrease in fatigue (OR = 0.4) and headache (OR = 0.2) was observed. An association between female gender and sinusitis was found before the remediation (OR = 8.1). Age over 40 years was a risk factor for voice problems and more than 10 working years at the same school were associated with increased risk for conjunctivitis (OR = 8.5) and headache (OR = 5.4). Other exposure situations such as mould problems at home and mould exposure during leisure time also have an effect on teachers' health. Significant reduction was found in symptoms of fatigue and headache after the cessation of exposure, while respiratory symptoms need much longer time to relieve after the remediation. Age, female gender, atopy, long-term exposure time and other exposure situations might be the risk factors for prolonged symptoms among mould exposed teachers. PMID- 15545039 TI - Microbiological quality of food samples from restaurants and sweet shops in developing countries: a case study from the Occupied Palestinian Territory. AB - The aim of this paper is to review the microbiological quality of food samples taken from a wide range of cooked and raw foods obtained from restaurants and sweet shops in the Ramallah and al-Bireh district, and to identify the gaps in food inspection and handling that can be realistically improved. Utilizing food sample test results of the Palestinian Ministry of Health, records pertaining to the years 1995, 1996, 2000, 2002, and the first 2 months of the year 2003 reveal that only 60.9%, 44.0%, 63.8%, 93.6%, 51.8%, 83.8%, and 50.4% of the food samples tested for total aerobic count, total Coliform, faecal Coliform, Staphylococcus aureus, faecal Streptococci, moulds, and yeasts respectively are within the limits of the Palestinian and International standards. None of the tested samples for Salmonellae or Clostridium perfringes were positive. Analysis of the data indicates an irregular rather than systematic process to testing. For example, 60.4% of the tested food samples in 2000 came from sweet shops, while the relative percentage of sweet shops is 21.4% out of the total restaurants and sweet shop figures in the area at the time. In contrast to what would be expected, most of the samples were collected during the cold season, raising questions as to the suitability of testing procedures and guidelines. Systemic and procedural gaps were identified by the analysis that can be addressed to at least contain, if not completely eliminate the presence of foods in the market that are unacceptable for consumption. PMID- 15545040 TI - Risk and management in hospital water systems for Legionella pneumophila: a case study in Rio de Janeiro-Brazil. AB - This article analyses the water used at hospitals in Rio de Janeiro, Brazil. The research, based on microbiological and physical-chemical aspects, suggests subsidies for normalization of hospital potable water systems and makes recommendations for standardization of operational procedures for inspection for Legionella pneumophila. A total of 16 hospitals were inspected and positive results for the presence of L. pneumophila were found at five hospitals. These hospitals were integrated in a research project aiming at the detection and quantification of this pathogen. During 10 consecutive weeks, four collections representing a total of 200 analyses were done at the five researched hospitals. In this way seven physical-chemical parameters and three microbiological parameters were observed to evaluate the quality of water in each hospital. The results showed that routine surveillance for a hospital water distribution system is fundamental for public health and must include, as a priority, monitoring of L. pneumophila. The water quality varies in accordance with the hospital water system involved. It is important and necessary to implement environmental culturing in order to minimize hospital infection, in particular, pneumonia data and also to provide the basis for disinfection of the water system. PMID- 15545041 TI - How clean is clean enough? Recent developments in response to threats posed by chemical and biological warfare agents,. PMID- 15545043 TI - Protection capacity against low-density lipoprotein oxidation and antioxidant potential of some organic and non-organic wines. AB - Current research suggests that phenolics from wine may play a positive role against oxidation of low-density lipoprotein (LDL), which is a key step in the development of atherosclerosis. Considering the effects of different wine-making techniques on phenols and the wine consumption preference influencing the benefical effects of the product, organically and non-organically produced wines were obtained from the grapes of Vitis vinifera origin var: Carignan, Cabernet Sauvignon, Merlot, Grenache, Columbard and Semillon. Levels of total phenols [mg/l gallic acid equivalents (GAE)], antioxidant activity (%) and inhibition of LDL oxidation [%, inhibition of diene and malondialdehyde (MDA) formation] were determined. Some phenolic acids (gallic acid, p-hydroxybenzoic acid, syringic acid, 2,3-dihydroxybenzoic acid, ferulic acid, p-coumaric acid and vanillic acid) were quantified by high-performance liquid chromatography equipped with an electrochemical detection carried at +0.65 V (versus Ag/AgCl, 0.5 microA full scale). The highest concentrations of gallic, syringic and ferulic acids were found in organic Cabernet Sauvignon; 2,3-dihydroxybenzoic acid in organic Carignan and p-coumaric and vanillic acids in non-organic Merlot wine. High levels of antioxidant activity (AOA), inhibition of LDL oxidation and total phenol levels were found in non-organic Merlot (101.950% AOA; 88.570% LDL-diene; 41.000% LDL-MDA; 4700.000 mg/l GAE total phenol) and non-organic Cabernet Sauvignon (92.420% AOA; 91.430% LDL-diene; 67.000% LDL-MDA; 3500.000 mg/l GAE total phenol) grape varieties. Concentrations of some individual phenolic constituents (ferulic, p-coumaric, vanillic) are correlated with high antioxidant activity and inhibition of LDL oxidation. The best r value for all examined characteristics was determined for gallic acid, followed by 2,3-dihydroxybenzoic, syringic, ferulic and p-coumaric acids. Negative correlation of vanillic with MDA and p-hydroxybenzoic acid with LDL were confirmed by principal component analysis (PCA) analyses. Red wines display a higher antioxidant activity (81.110% AOA) than white ones (19.512% AOA). The average level of LDL inhibition capacity in red wine was determined as 87.072% and for the white as 54.867%. PMID- 15545044 TI - Nutritional status of cancer patients given different treatment modalities. AB - The nutritional status of 91 cancer patients was assessed at the time of diagnosis and follow-up assessments were carried out at the third and sixth week after initiating different treatment modalities to study the effect of type and duration of treatment on nutritional status. Parameters assessed were anthropometry, biochemical status and clinical signs and symptoms of nutritional deficiencies. Treatment modalities studied were radiotherapy, chemotherapy, chemotherapy+radiotherapy, and combined treatment modality (surgery+radiotherapy+chemotherapy). The nutritional status of male patients was affected most by chemotherapy+radiotherapy while females were affected most with radiotherapy. Biochemical parameters showed a marginal decline in total serum protein and serum albumin concentrations. Haemoglobin concentrations declined substantially with radiotherapy and chemotherapy. The lymphocyte count decreased substantially irrespective of the treatment modality. Clinical examination revealed increased incidences of deficiency signs and symptoms in all patients during follow-up irrespective of treatment modality. PMID- 15545045 TI - Fat and sodium chloride reduction in sausages using kappa-carrageenan and other salts. AB - Fat and sodium chloride were reduced in a sausage formulation including kappa carrageenan and other salts, as potassium and calcium chloride, in different concentrations, in order to compensate the ionic strength during myofibrillar protein extraction and solubilisation, and to promote the carrageenan stable conformation and gelation. Four different treatments were employed reducing fat from 15 to 10% and sodium chloride from 2.5 to 1.5% and 1%. Potassium chloride was added at 0.5% to all the treatments, and calcium chloride to 0.5% and 0.01% in the last two. The cooking yield was higher for all the treatments but expressible moisture was not significantly different, meaning that the water is not chemically entrapped by carrageenan at the ionic strength conditions employed. However, since no detrimental cooking losses or fat release were detected, myofibrillar proteins maintain a good functionality at these conditions together with kappa-carrageenan. Low-fat sodium-reduced treatment results were slightly darker but redder than the control, probably due to less fat in the formulation. Textural profile analysis demonstrated that, at the fat level employed and the different sodium, potassium and calcium chloride concentrations, similar textures could be created. Sensory analysis indicated that from the formulations employed, 1.5% NaCl with 0.5% KCl and 0.01% CaCl(2) was similar in controlling flavour (juiciness) and texture (hardness). These results established the possibility to reduce significantly the fat and sodium chloride content employing kappa-carrageenan with potassium and calcium chloride, without detrimental effects on texture and sensory characteristics. PMID- 15545046 TI - Nutritional status of 8-year-old rural and urban Italian children: a study in Pistoia, Tuscany. AB - In this study, performed in the province of Pistoia (Italy), we tested whether 8 year-old children living in rural areas differed from their urban peers as far as nutritional status, dietary habits and physical activity are concerned. The study sample was randomly selected to include 50% of the children attending the third elementary class in the province of Pistoia during 2002. A total of 1006 children underwent an anthropometric evaluation and an assessment of dietary habits and physical activity by means of specific questionnaires. Of these children, 927 were born in Italy and are described in this report. The relationship between body mass index (BMI<5th percentile versus 5thor=95th percentile) and the environment (urban versus rural) was of borderline significance (P=0.051). However, the percentage of children with BMI>or=85th percentile was greater in rural (24%) than in urban (18%) areas (P=0.012). The choice of foods and the physical activity of rural children mirrored those of urban children. These data sound as an alarm in view of the current epidemics of obesity that are invading Italy and other Mediterranean countries. PMID- 15545047 TI - The effect of meat products enriched with plant sterols and minerals on serum lipids and blood pressure. AB - The purpose of the study was to investigate the effect of non-esterified plant sterol-enriched and mineral-enriched low-fat and low-salted meat products compared with control meat products, on serum total and lipoprotein lipids and blood pressure in subjects with mildly to moderately elevated serum cholesterol concentration. A randomised, placebo-controlled, single-blind, repeated measure design was used. Altogether 21 volunteers completed the study. The study began with a pre-trial period of 1-2 weeks, which was followed by three different test periods in the following order: meat products enriched with plant sterols (1.2 g/day), potassium, calcium and magnesium (MP1); meat products with no added plant sterols and minerals (control); and meat products with plant sterols (2.1 g/day), potassium, calcium and magnesium (MP2). Each test period lasted for 3 weeks. During the MP2 period, the serum total and low-density lipoprotein cholesterol concentration decreased 4.9+/-7.5% (P<0.05) and 4.6+/-11.3% (not significant), respectively, compared with the control period. No differences in the high density lipoprotein cholesterol and total triglyceride concentrations or in systolic blood pressure and diastolic blood pressure were found among the test periods. In conclusion, the present study showed that frankfurters and cold cuts enriched with plant sterols from tall oil, potassium, calcium and magnesium, as part of habitual Finnish diet reduced the serum total cholesterol concentration in hypercholesterolemic subjects when the intake of sitosterols was 2.1 g/day, but not with the lower dose. PMID- 15545048 TI - Resistant starch derived from processed legumes: in vitro and in vivo fermentation characteristics. AB - The effect of processing of legumes on resistant starch formation, its rate of fermentation and the production of short chain fatty acids under in vitro and in vivo systems was assessed. The content of resistant starch in pressure-cooked Bengal gram, black gram and red gram was 3.59%, 1.58% and 3.34%, respectively. Fermentation in vitro of resistant starch derived from processed red gram showed higher amount of short chain fatty acids (2.38 mmol), especially butyric acid (2.22 mmol). Under in vivo conditions (in albino rats) all processed legumes showed a higher faecal bulking, and more short chain fatty acids, with a significant increase in the anaerobic bacterial counts. Compared with a processed legume diet, the caecum of animals fed a raw diet showed a preponderance of propionic acid. PMID- 15545049 TI - Nutritional and sensory quality of stirred soursop (Annona muricata L.) yoghurt. AB - Soursops (Annona muricata L.) are highly aromatic fruits with white juicy flesh and are native to tropical North and South America. The ripe fruits are highly perishable, as they become soft and easily bruised. The objectives of the study were to incorporate soursop nectar at 0%, 5%, 10% and 15% in stirred yoghurts and to analyse the products for chemical and sensory quality. A focus group evaluated the initial yoghurts for process modifications. Yoghurts were evaluated on sensory attributes of appearance and colour, body and texture, flavour and aroma, and overall quality. Yoghurts with 10% and 15% soursop nectar had the highest (P<0.05) overall quality scores (12.60/20 and 12.75/20, respectively) but differed (P<0.05) in flavour and aroma from plain yoghurt and 5% soursop yoghurt. Most panelists would consider purchase of 10% and 15% soursop yoghurts over 0% and 5% soursop yoghurts. These yoghurts provided high percentage daily values of zinc, phosphorus and calcium and a good level of protein. PMID- 15545050 TI - A study on degradation kinetics of ascorbic acid in amla (Phyllanthus emblica L.) during cooking. AB - The kinetics of ascorbic acid degradation in amla (Phyllanthus emblica L.) as well as in pure ascorbic acid solutions at initial concentrations present in amla over a temperature range of 50-120 degrees C (steady-state temperature) has been studied. The ascorbic acid degradation followed first-order reaction kinetics where the rate constant increased with an increase in temperature. The temperature dependence of degradation was adequately modeled by the Arrhenius equation. The activation energies were found to be 4.09 kcal/mole for amla and 4.49 kcal/mole for pure vitamin solution. The degradation kinetics of ascorbic acid was also evaluated in normal open pan cooking, pressure-cooking and a newly developed and patented fuel-efficient EcoCooker (unsteady state heating process). A mathematical model was developed using the steady-state kinetic parameters obtained to predict the losses of ascorbic acid from the time-temperature data of the unsteady state heating processing method. The results obtained indicate the ascorbic acid degradation is of a similar order of magnitude in all the methods of cooking. PMID- 15545051 TI - Antioxidant activities of various fruits and vegetables produced in Taiwan. AB - Fruits and vegetables have been known to contain a variety of antioxidant components. It has been suggested that antioxidants may protect biomolecules from oxidative damage and therefore be associated with reduced risks of cardiovascular disease and certain cancer. The antioxidant abilities of various parts of eight common fruits and vegetables produced in Taiwan were investigated, including tomato, guava, squash, tangerine, wax gourd, pineapple, chayote, and eggplant. Squash, wax gourd, tomato, and guava seeds showed the highest antioxidant activities in thiobarbituric acid assay. Wax guard and squash seeds showed the highest antioxidant activities in iodometric assay. At the level of 1 g fresh sample, low-density lipoprotein peroxidation was inhibited by at least 90% by tomato meat, guava meat, squash seed, wax gourd meat, core, and seed, and eggplant skin. The total phenolic content was significantly correlated with antioxidant activities measured by thiobarbituric acid (r=0.715, P<0.01) and iodometric (r=0.749, P<0.01) assays. The results of this study could be used for development of merchandise with potential health benefits from agricultural products. PMID- 15545052 TI - Application of response surface methodology for studying the product characteristics of extruded rice-cowpea-groundnut blends. AB - Response surface methodology (with central composite rotatable design for k=3) was used to investigate the product properties of extruded rice-cowpea-groundnut blends in a single screw extruder. The combined effect of cowpea (0-20%), groundnut (0-10%), and feed moisture (14-48%) levels were used for formulation of the products. The product moisture, expansion ratio, bulk density and total colour change were studied using standard analytical methods. Well-expanded rice legume blend extrudates of less bulk density and lower moisture content were produced at low feed moisture. Increasing legume addition affected the various shades of colour in the product. Models developed for the indices gave R(2) values ranging from 52.8% (for the b-value) to 86.5% (for bulk density). The models developed suggested that the optimal process variables for the production of a puffed snack with an enhanced nutrition and spongy structure from a rice cowpea-groundnut blend are low feed moisture of 14-20% and maximum additions of 20% cowpea and 10% groundnut. A lack-of-fit test showed no significance, indicating that the models adequately fitted the data. PMID- 15545053 TI - Circulating levels of cyclooxygenase metabolites in experimental Trypanosoma cruzi infections. AB - Trypanosoma cruzi induces inflammatory reactions in several tissues. The production of prostaglandin F2alpha, 6-keto-prostaglandin F1alpha and thromboxane B2, known to regulate the immune response and to participate in inflammatory reactions, was studied in mice experimentally infected with T. cruzi. The generation of nitric oxide (NO), which could be regulated by cyclooxygenase metabolites, was also evaluated. In the acute infection the extension of inflammatory infiltrates in skeletal muscle as well as the circulating levels of cyclooxygenase metabolites and NO were higher in resistant C3H mice than in susceptible BALB/c mice. In addition, the spontaneous release of NO by spleen cells increased earlier in the C3H mouse strain. In the chronic infections, the tissue inflammatory reaction was still prominent in both groups of mice, but a moderate increase of thromboxane B2 concentration and in NO released by spleen cells was observed only in C3H mice. This comparative study shows that these mediators could be mainly related to protective mechanisms in the acute phase, but seem not to be involved in its maintenance in the chronic T. cruzi infections. PMID- 15545054 TI - The limited immunomodulatory effects of escharectomy on the kinetics of endotoxin, cytokines, and adhesion molecules in major burns. AB - Escharectomy has been shown to improve the survival rates and the outcomes in burns. This observational study was conducted to assess the role of escharectomy on the inflammatory mediators in major burns. Seventeen ASA physical status II or status III adult surviving major burn patients were recruited. When the escharectomy was scheduled, a series of blood samples was obtained at -3 and -1 days preoperation, and +1 and +3 postoperation. The changing levels of endotoxin, cytokines, and adhesion molecules were measured with a quantitative sandwich immunoassay. Extensive escharectomy did not appear to have any significant impact on the levels of tumor necrosis factor alpha, interleukin-10, soluble intracellular adhesion molecule-1 and soluble vascular adhesion molecule-1. Meanwhile, endotoxin and E-selectin were significantly decreased after escharectomy. Escharectomy appeared to have a limited immunomodulatory effect on the inflammatory mediators in systemic inflammatory responses induced by major burns. This is probably related to the timing and extent of surgery, and the complex nature of burn-related inflammation. PMID- 15545055 TI - Tc1/Tc2 ratio in the inflammatory process in patients with Behcet's disease. AB - BACKGROUND: Peripheral blood CD8+ T cells expressing interferon gamma and interleukin-4 (IL-4), and lacking CD28 molecules, were responsible for the dynamic interplay between peripheral blood and inflammatory sites. INTRODUCTION: The aim of the current study was to define in Behcet's disease (BD), CD8+ T-cell subsets using CD28 and CD11b monoclonal antibodies, and the characterization of the Tc1/Tc2 ratio and perforin expression. METHODS: Flow cytometry was used for intracytoplasmic cytokines and perforin expression. Effector cells were investigated by adhesion of CD8+ T cells to human microvascular endothelial cells and by chemotaxis using beta-chemokine. RESULTS: Interferon-gamma-producing CD8+ T cells in active and remission BD patients were increased, which induce a significant increase of the Tc1:Tc2 ratio in BD. CD8(+)CD28(-)CD11b+ T cells were found to be more expanded in BD patients than in age-matched healthy controls. The expression of CD11b molecules in active BD allowed to CD8(+)CD28+/CD8(+)CD28- subsets to adhere to human microvascular endothelial cells, with more efficiency in BD. Using MIP-1alpha, we observed that the migratory process of CD28( )CD11b(+) is more important in BD. CD28(-)CD11b+ exhibited an increased perforin expression in BD patients. CONCLUSION: Taken together these results suggest the presence of immune activation, probably in response to a profound inflammation affecting BD patients. The physiopathological significance of these results were toward autoimmune diseases and/or infectious process. PMID- 15545056 TI - Differential activation of NF kappa B/RelA-p50 and NF kappa B/p50-p50 in control and alcohol-drinking rats subjected to carrageenin-induced pleurisy. AB - BACKGROUND: Carrageenin (CAR) injection into the pleural cavity causes local inflammation called carrageenin-induced pleurisy (CAR-IP). Inflammation onset is characterized by an activation of pro-inflammatory NFkappaB, RelA-p50, while inflammation resolution is characterized by an activation of an anti-inflammatory NFkappaB, p50-p50, that re-establishes homeostasis, an essential process for an organism's survival. Although chronic alcohol intake disrupts inflammation, the mechanism behind the development of inflammatory disorder in alcoholics is not yet known. Therefore, the aim of this investigation was to study the effects of ethanol intake on CAR-IP and NFkappaB activation in pleural fluid neutrophils in P rats. METHODS: Alcohol-preferring, P rats were given free choice of alcohol (15% ethanol) and water or water alone (for control) for 15 days. Then, each rat was injected with 0.2 ml of 2% CAR into the pleural cavity under light ether anesthesia. At different time intervals after the CAR injection, rats were anesthetized and their blood and pleural fluid samples were collected. Pleural fluid inflammatory cells were identified with Turk's or Wright-Giemsa staining. Different cell types were sorted using a fluorescence-activated cell sorter. Pleural fluid neutrophils were examined for apoptosis and activation of the two NFkappaB subspecies. RESULTS: In control rats, fluid began to accumulate in the pleural cavity 0.5 h after, which peaked 24 h after, CAR injection. Then, the values declined gradually. The increase in pleural fluid correlated with RelA-p50 activation, while the decline in pleural fluid correlated with p50-p50 activation and apoptosis in neutrophils. In alcohol-drinking rats, pleural fluid remained elevated for up to 6 days after CAR injection. Neutrophils from alcohol-drinking rats exhibited suppressed apoptosis, augmented RelA-p50 activation, and suppressed p50-p50 activation. CONCLUSIONS: Alcohol intake prolonged inflammation in P rats. An alcohol-induced upregulation of RelA-p50 activation and downregulation of p50-p50 activation may be causally related to the alcohol induced inflammation dysregulation. PMID- 15545057 TI - Effects of 'casoparan', a peptide isolated from casein hydrolysates with mastoparan-like properties. AB - Casein, a protein found in milk of several species, is divided into different chains from 19 to 25 kDa. Casein is also considered as a source of amino acids and generating peptides with biological activities such as opiate, immunostimulating, antibacterial, peptidase inhibitors, among others. In this work, Sephadex G-10 chromatography followed by high-performance liquid chromatography isolation purified NZCase TT, an industrial culture media for tetanus toxin production. In the first step, four pools were isolated and tested in different assays: isolated smooth muscle assay (guinea pig ileum, rat uterus), phagocytosis in vitro of opsonized sheep red blood cells, and hydrogen peroxide (H2O2) release from mouse peritoneal macrophages. Pool III was the main active pool being able to potentiate bradykinin action in guinea pig ileum, stimulating phagocitic activity by resident macrophages and increasing H2O2 release from macrophages previously activated with bacille Calmette Guerin. Using mass spectra the primary structure of the main peptide from pool III was obtained--INKKI, which corresponds to beta-casein fragment 26-30. The immunostimulating action is probably related to a direct action in macrophage cells. PMID- 15545058 TI - Serum procalcitonin and cerebrospinal fluid cytokines level in children with meningitis. AB - AIMS: To determine the level of serum procalcitonin and cerebrospinal fluid cytokines in children with bacterial or viral meningitis and to document the use of these parameters in differential diagnosis. RESULTS: Before the start of antibiotic treatment, serum procalcitonin and tumor necrosis factor alpha levels were found to be higher in acute bacterial meningitis compared with viral meningitis and with the control group. Similarly, cerebrospinal fluid interleukin 6 levels were found to be significantly higher in children with acute bacterial meningitis compared with viral meningitis. However, no significant difference was determined between groups in respect to the cerebrospinal fluid interleukin-8 level. CONCLUSION: Serum procalcitonin and cerebrospinal fluid tumor necrosis factor alpha levels can be used in the early diagnosis of bacterial meningitis. Similarly, they may be useful adjuncts in differential diagnosis of bacterial and viral meningitis. PMID- 15545060 TI - Inhaled corticosteroid effects both eosinophilic and non-eosinophilic inflammation in asthmatic patients. AB - AIM: To determine induced sputum cell counts and interleukin-8 (IL-8), tumor necrosis factor alpha (TNF-alpha) and leukotriene B4 (LTB4) levels as markers of neutrophilic inflammation in moderate persistent asthma, and to evaluate the response to inhaled steroid therapy. METHODS: Forty-five moderate asthmatic patients and 10 non-smoker controls were included in this study. All patients received inhaled corticosteroid (800 microg of budesonide) for 12 weeks. Before and after treatment pulmonary function tests were performed, and symptom scores were determined. Blood was drawn for analysis of serum inflammatory markers, and sputum was induced. RESULTS: Induced sputum cell counts and inflammatory markers were significantly higher in patients with asthma than in the control group. The induced sputum eosinophil counts of 12 patients (26%) were found to be less than 5%, the non-eosinophilic group, and sputum neutrophil counts, IL-8 and TNF-alpha levels were significantly higher than the eosinophilic group (neutrophil, 50+/ 14% versus 19+/-10%, p<0.01). In both groups, there was a significant decrease in sputum total cell counts and serum and sputum IL-8, TNF-alpha and LTB4 levels after the treatment. There was no change in sputum neutrophil counts. Although the sputum eosinophil count decreased only in the eosinophilic subjects, there was no significant difference in inflammatory markers between the groups. The symptom scores were significantly improved after treatment, while the improvement did not reach statistical significance on pulmonary function test parameters. CONCLUSION: Notably, in chronic asthma there is a subgroup of patients whose predominant inflammatory cells are not eosinophils. Sputum neutrophil counts and neutrophilic inflammatory markers are significantly higher in these patients. In the non-eosinophilic group, inhaled steroid caused an important decrease in inflammatory markers; however, there was no change in the sputum eosinophil and neutrophil counts. PMID- 15545059 TI - Circulating cytokine profile in anti-neutrophilic cytoplasmatic autoantibody associated vasculitis: prediction of outcome? AB - AIMS: The anti-neutrophilic cytoplasmatic autoantibody-associated vasculitides (AASV) are diseases of relapsing-remitting inflammation. Here we explore the cytokine profile in different phases of disease, looking for pathogenic clues of possible prognostic value. RESULTS: Interleukin (IL)-6, IL-8 and IL-10 were significantly elevated in plasma. Patients in the stable phase who subsequently developed adverse events had higher IL-8 values. Patients in the stable phase who relapsed within 3 months had lower IL-10 values and higher IL-6 levels. CONCLUSIONS: Patients with AASV have raised circulating cytokine levels compared with healthy controls, even during remission. Raised IL-8 seems associated with poor prognosis. Lower levels of IL-10 and higher levels of IL-6 herald a greater risk of relapse. Patients with systemic vasculitis in clinical remission have persistent disease activity, kept under control by inhibitory cytokines. PMID- 15545062 TI - Bacteriophage Phi S1 infection of Pseudomonas fluorescens planktonic cells versus biofilms. AB - This communication focuses on the efficacy of a specific lytic phage, phage Phi S1, as a control agent of Pseudomonas fluorescens biofilms. The effect of phage infection temperature and the host growth temperature were evaluated. The results obtained showed that the phage infection process was temperature dependent and that the optimum temperature of infection of planktonic cells and biofilms was 26 degrees C. At this temperature, bacteriophage Phi S1, at a multiplicity of infection (MOI) of 0.5 infected both planktonic cells and biofilms causing a biomass reduction of about 85% in both cases. PMID- 15545061 TI - Induction of macrophage migration inhibitory factor precedes the onset of acute tonsillitis. AB - We investigated the serum macrophage migration inhibitory factor (MIF) levels of palmoplantar pustulosis patients, before and after the tonsillar provocation test. Higher serum MIF levels of palmoplantar pustulosis patients were decreased after the tonsillar provocation test (n=29). To confirm these phenomena, two patients with acute tonsillitis had their changes in body temperature, C-reactive protein (CRP) and serum MIF levels examined during the course of their illness. Surprisingly, increased MIF preceded fever and CRP elevation, and MIF subsequently decreased at the onset of fever and CRP elevation. Since MIF is an initiator of other proinflammatory cytokines, we suggest that the induction of MIF may precede other inflammatory conditions. PMID- 15545063 TI - Isethionic acid and floridoside isolated from the red alga, Grateloupia turuturu, inhibit settlement of Balanus amphitrite cyprid larvae. AB - Isethionic acid (2-hydroxyethane sulfonic acid) and floridoside (2-O-alpha-D galactopyranosylglycerol) were extracted from the red alga, Grateloupia turuturu, and tested for anti-settlement activity against cyprid larvae of the tropical barnacle, Balanus amphitrite and for their toxicity to nauplius larvae. Isethionic acid was active for anti-settlement but had the disadvantage of being toxic to nauplius larvae. Floridoside was a potent inhibitor of cyprid settlement at non-toxic concentrations to nauplii (0.01 mg ml(-1)). PMID- 15545064 TI - The impact of desiccation on the adhesion of barnacles attached to non-stick coatings. AB - Fouling-release coatings prevent fouling of ships' hulls through hydrodynamic forces generated as the ship moves through the water. The effectiveness of such coatings may be evaluated by measuring the adhesion strength of settled organisms, e.g. barnacles. The influence of desiccation of the barnacle adhesive on such measurements was investigated. Shear forces required to remove barnacles of the genus Balanus increased during the course of desiccation up to the point when the barnacles suddenly self-detached. The increase was thought to be due to the rising cohesive strength of the adhesive. Growing tensile forces within the weakly cross-linked adhesive, however, are suggested to have led to self detachment. The shear forces required to remove barnacles of the genus Elminius were generally low and did not differ significantly during the course of desiccation. The different results may be attributed to specific base morphologies. It was concluded that measuring the adhesion strength of members of the Balanidae on non-stick surfaces in air could produce flawed results due to the influence of desiccation of the barnacle adhesive. The investigations have also provided new insights into the characteristics of barnacle adhesive. PMID- 15545065 TI - Abundance of macroalgal organic matter in biofilms: evidence from n-alkane biomarkers. AB - Biofilm development on titanium panels immersed in the surface waters of Dona Paula Bay was investigated using molecular biomarkers such as n-alkanes and other chemical and biological parameters. Biofilm biomass measured as organic carbon (OC), organic nitrogen (ON), chlorophyll a, diatoms and bacterial numbers on the titanium panels generally increased over the period of immersion. Total lipids and n-alkane concentration also showed similar trends. n-alkanes from C(12) to C(30) were detected in the biofilm samples, which showed a bimodal distribution. The first mode consisted of n-alkanes > C(23) with a strong even over odd predominance. In the second mode, the n-alkanes < C(23) were more abundant with odd carbon number maxima at C(15), C(17) and C(19) and a strong odd over even carbon number predominance (Carbon Preference Index > 2). The predominance of these odd-chain n-alkanes strongly indicates that the organic matter derived from macroalgal sources was the major contributor to the biofilm organic matter developed on the titanium panels over the 15 d period of study. The data suggest that molecular characterization is a useful tool in understanding the sources of biofilm organic matter. The observed abundance of macroalgal organic matter during the 15 d period of biofilm development may play an important role in subsequent fouling by micro- and macrofouling organisms. PMID- 15545066 TI - Photocatalytic inhibition of microbial adhesion by anodized titanium. AB - Biofouling is one of the concerns in the use of titanium for seawater cooled condensers of power plants. Earlier studies have shown that anodized titanium and its alloys with a thin film of anatase (TiO(2)) on its surface can inhibit attachment of Pseudomonas sp. when illuminated with near-UV light (350 - 380 nm). In the present study, a comparison of the photocatalytic inhibition of microbial attachment on titanium surfaces anodized at different voltages was carried out. Thin films of anatase of varying thickness were produced on titanium grade-2 by anodizing in dilute orthophosphoric acid solution at 30 V, 50 V and 100 V. The photocatalytic efficiency of these anodized surfaces was measured by the methylene blue degradation method. The anodised surfaces were exposed to liquid cultures of Gram-negative Pseudomonas sp., Gram-positive Micrococcus sp. and to a mixed algal culture. Photocatalytic inhibition of microbial attachment was maximum on the titanium surface anodized at 30 V, followed by the surface anodized at 50 V and then at 100 V. The photocatalytic inhibition of microbial attachment was also found to be dependent on the cell wall characteristics of the organism. The Gram-negative Pseudomonas sp. with a lipoproteinaceous outer membrane was the most susceptible to the photocatalytic effect, while the Gram positive Micrococcus sp. with peptidoglycan cell wall showed moderate susceptibility and the algae with siliceous cell wall showed no susceptibility at all. PMID- 15545067 TI - The effect of depth on the accrual of marine biofilms on glass substrata deployed in the Clyde Sea, Scotland. AB - Recent concerns about fouling problems caused by biofilms affecting optical oceanographic instruments have highlighted the need for a better understanding of their nature and extent in the marine environment. Glass slides were deployed in April and August for periods of up to 3 weeks at 5, 10, 20, 40, 80 and 160 m in the water column of Loch Fyne, Clyde Sea, W Scotland. Biofilms were enumerated using epifluorescence and bright field microscopy. During April the biofilm community varied significantly with depth, although this effect was attributable solely to changes in the diatom community. Diatom numbers peaked between 10 and 20 m. During August the biofilm community also showed a significant depth effect, although in this case there were significant effects for diatoms, rods, and filamentous bacteria. Cell numbers for diatoms, filamentous bacteria, and rod shaped bacteria peaked at 5 m. There was a significant linear relationship between the number of diatoms and bacteria on the slides deployed in August. No such relationship was found for the April data. The results indicate that optical performance may be significantly degraded after a few weeks and highlights the need for provision of suitable strategies to protect such surfaces from biofilm accumulation. PMID- 15545068 TI - Characterisation of structures in salivary secretion film formation. An experimental study with atomic force microscopy. AB - The purpose of the present study was to characterise the structure dynamics of pure salivary secretions retained on controlled surfaces with different surface energies in the early stage of salivary film formation. Germanium prisms prepared to have either low surface energy or medium surface energy were incubated in fresh secretions of either human parotid saliva (HPS) or human submandibular/sublingual saliva (HSMSLS) for 15, 90, and 180 min. After controlled rinsing with distilled water, the surfaces were air dried and thereafter imaged with atomic force microscopy (AFM). The amount of adsorbed material and the size of the structures detected increased with increased saliva exposure time. The film thicknesses varied from 10 to 150 nm, and both HPS and HSMSLS films contained structures with diameters varying from 40 nm to 2 microm. Some of these were clustered into special formations. The HPS films exhibited a more granular morphology than the HSMSLS films. Furthermore, branched lines were detected on the low surface energy germanium prisms incubated in saliva. The results indicate that exposure time, surface energy, and type of salivary secretion all are factors affecting the adsorption characteristics of salivary films. PMID- 15545069 TI - Effect of electronic stability control on automobile crash risk. AB - Per vehicle crash involvement rates were compared for otherwise identical vehicle models with and without electronic stability control (ESC) systems. ESC was found to affect single-vehicle crashes to a greater extent than multiple-vehicle crashes, and crashes with fatal injuries to a greater extent than less severe crashes. Based on all police-reported crashes in 7 states over 2 years, ESC reduced single-vehicle crash involvement risk by approximately 41 percent (95 percent confidence limits 3348) and single-vehicle injury crash involvement risk by 41 percent (2752). This translates to an estimated 7 percent reduction in overall crash involvement risk (310) and a 9 percent reduction in overall injury crash involvement risk (314). Based on all fatal crashes in the United States over 3 years, ESC was found to have reduced single-vehicle fatal crash involvement risk by 56 percent (3968). This translates to an estimated 34 percent reduction in overall fatal crash involvement risk (2145). PMID- 15545070 TI - Effectiveness of mandatory license testing for older drivers in reducing crash risk among urban older Australian drivers. AB - Most licensing jurisdictions in Australia maintain mandatory assessment programs targeting older drivers, whereby a driver reaching a specified age is required to prove his or her fitness to drive through medical assessment and/or on-road testing. Previous studies both in Australia and elsewhere have consistently failed to demonstrate that age-based mandatory assessment results in reduced crash involvement for older drivers. However studies that have based their results upon either per-population or per-driver crash rates fail to take into account possible differences in driving activity. Because some older people maintain their driving licenses but rarely if ever drive, the proportion of inactive license-holders might be higher in jurisdictions without mandatory assessment relative to jurisdictions with periodic license assessment, where inactive drivers may more readily either surrender or lose their licenses. The failure to control for possible differences in driving activity across jurisdictions may be disguising possible safety benefits associated with mandatory assessment. The current study compared the crash rates of drivers in Melbourne, Australia, where there is no mandatory assessment and Sydney, Australia, where there is regular mandatory assessment from 80 years of age onward. The crash rate comparisons were based on four exposure measures: per population, per licensed driver, per distance driven, and per time spent driving. Poisson regression analysis incorporating an offset to control for inter jurisdictional road safety differences indicated that there was no difference in crash risk for older drivers based on population. However drivers aged 80 years and older in the Sydney region had statistically higher rates of casualty crash involvement than their Melbourne counterparts on a per license issued basis (RR: 1.15, 1.02-1.29, p=0.02) and time spent driving basis (RR: 1.19, 1.06-1.34, p=0.03). A similar trend was apparent based on distance travelled but was of borderline statistical significance (RR: 1.11, 0.99-1.25, p=0.07). Collectively, it can be inferred from these findings that mandatory license re-testing schemes of the type evaluated have no demonstrable road safety benefits overall. Further research to resolve this on-going policy debate is discussed and recommended. PMID- 15545071 TI - Adjusting for car occupant injury liability in relation to age, speed limit, and gender-specific driver crash involvement risk. AB - It is well established that older drivers' fragility is an important factor associated with higher levels of fatal crash involvement for older drivers. There has been less research on age-related fragility with respect to the sort of minor injuries that are more common in injury crashes. This study estimates a quantity that is related to injury fragility: the probability that a driver or a passenger of that driver will be injured in crashes involving two cars. The effects of other factors apart from drivers' fragility are included in this measure, including the fragility of the passengers, the crashworthiness of cars driven, seatbelt use by the occupants, and characteristics of crashes (including configuration and impact speed). The car occupant injury liability estimates appropriately includes these factors to adjust risk curves by age, gender, and speed limit accounting for overrepresentation in crashes associated with fragility and these other factors. PMID- 15545072 TI - Road rage experience and behavior: vehicle, exposure, and driver factors. AB - Road rage has generated increasing public concern. Research has shown that victimization and perpetration of road rage is more common among males and younger drivers. We aimed to extend the understanding of determinants of road rage to driving exposure and vehicle factors, based on a 20022003 population survey of 1,631 regular drivers in Ontario, Canada. Regression analyses revealed that number of times drivers reported experiencing road rage in the previous 12 months was significantly greater for males, younger respondents, and those residing in Toronto. Also, victimization was significantly greater for drivers who did all their driving on busy roads and increased with number of kilometers driven on a typical week; however, type of vehicle driven was not significant. Number of times road rage perpetration was reported in the past 12 months was significantly greater for males, younger respondents, and those residing in Toronto, and lower for those in the Eastern and Northern region. Road rage perpetration increased significantly with number of weekly kilometers driven and was significantly greater for drivers who are always on busy roads and lower for those who never drive on busy roads, and higher for high-performance vehicle drivers. Even after controlling for driving exposure, road rage victimization and perpetration were highest for drivers in Toronto, where the pace of life may be more demanding. As expected, high-performance vehicle drivers reported more road rage perpetration. These individuals may experience more frustration when they are prevented from using the full performance capacities of their vehicles by crowded urban roadways. PMID- 15545073 TI - Reliability and validity of aggressive driving measures in China. AB - The present research was designed to examine the psychometric properties of Chinese versions of the Self Report Driver Behavior Aggression and Assertiveness subscales, the Driving Vengeance Questionnaire, and the Violent Driving Questionnaire. Study 1 found that the all scales demonstrated good internal consistency, with alphas ranging from .76 to .87 and that assertive driving was related to demerit points received over the past 12 months while driver aggression and violence were linked to collisions over the past 12 months. Study 2 found that the scales exhibited reasonable test-retest reliability, with correlations ranging from .82 to .89. Finally, Study 3 showed that each scale was predicted by other dangerous driving attitudes and behaviors, similar to the original versions. The consistency between the translated and original scales, the implications for use in a Chinese sample, and the uniformity of actions in the traffic environment across cultures are discussed. PMID- 15545074 TI - Tennessee's Novice Driver Safety Project: a program to increase parental involvement. AB - Graduated licensing has been implemented in many U.S. states to reduce teenage driver crash involvement. The goal is to introduce teenagers gradually to driving before allowing full, unrestricted licensure. Tennessee, one of the states that introduced graduated licensing, implemented a program to influence both driving by teenagers during the learner stage and restrictions imposed by parents after licensure. In addition to a standard welcome letter, booklets and reminder cards were mailed to parents of teenagers who had just obtained their learner's permits. The booklets and cards were designed to assist parents in becoming more actively involved in their children's driving experiences. The effects of three different approaches (welcome letter only, letter plus booklet, and letter plus booklet plus reminder cards) on parental involvement, teenage crashes, and teenage citations were assessed based on telephone interviews with parents. Although the parents said the booklets and reminder cards were helpful and should continue to be sent to other parents, there were no discernible effects on teenage practice driving and reported parental involvement during the learner stage. Nor was there any measurable influence on restrictions imposed by parents after licensure. It is unknown whether the standard welcome letter affected parental involvement, but overall supervised driving exceeded the state requirements by a substantial margin. PMID- 15545075 TI - Utilization of state-wide hospital separations data and the injury impairment scale to assess the incidence of spinal cord injury arising from motor vehicle traffic crashes. AB - The objectives of this study were to determine whether routine state-wide hospitalization data can be used for population level surveillance of the incidence of spinal cord injury (SCI) from motor vehicle traffic crashes (MVTCs) and to verify the coverage of the Australian Spinal Cord Injury Register. A method was developed to identify new injury incidents from routine South Australian hospitalization data. Mapping software was then used to derive the Abbreviated Injury Scale (AIS) and Injury Impairment Scale (IIS) codes for each incident case of spinal injury using the principal diagnosis code on the hospital data file. IIS code values in the range from 3 to 6 were considered to be comparable with the SCI coding criteria utilized by the Australian Spinal Cord Injury Register (ASCIR) which were based on the criteria of the American Spinal Injury Association. The number of estimated new incident cases of SCI based on the IIS coding of hospitalization data was compared with the actual number reported from the ASCIR. The case numbers were highly comparable overall and also by time and age group suggesting that the coverage of the ASCIR was very high. The results of the study support use of the IIS for estimating incident case numbers of MVTC-related SCI from routine state-wide hospitalization data. If routine hospital separations data elsewhere were suitable for the purpose and the IIS also proves to be applicable to non-MVTC-related SCI, the incidence of SCI could be monitored in state and national populations using the IIS and this could provide an alternative data source where registers of SCI do not exist. In addition, the IIS provides a means to assess coverage where registers do exist. PMID- 15545076 TI - Force and acceleration corridors from lateral head impact. AB - This study was conducted to provide force and acceleration corridors at different velocities describing the dynamic biomechanics of the lateral region of the human head. Temporo-parietal impact tests were conducted using specimens from ten unembalmed post-mortem human subjects. The specimens were isolated at the occipital condyle level, and pre-test x-ray and computed tomography images were obtained. They were prepared with multiple triaxial accelerometers and subjected to increasing velocities (up to 7.7 m/s) using free-fall techniques by impacting onto a force plate from which forces were recorded. A 40-durometer padding (50-mm thickness) material covering the force plate served as the impacting boundary condition. Computed tomography images obtained following the final impact test were used to identify pathology. Four specimens sustained skull fractures. Peak force, displacement, acceleration, energy, and head injury criterion variables were used to describe the dynamic biomechanics. Force and acceleration responses obtained from this experimental study along with other data will be of value in validating finite element models. The study underscored the need to enhance the sample size to derive probability-based human tolerance to side impacts. PMID- 15545077 TI - Evaluation criteria for AIS 1 neck injuries in frontal impacts--a parameter study combining field data and Madymo modeling. AB - Two situations with an expected higher AIS 1 neck injury rate in frontal impact were compared to a reference situation using a Madymo human body model in three different sitting postures and four different crash pulses. The two situations were reduced occupant weight and occupant with initial forward arm resistance, respectively. Occupant neck motion phases were identified and corresponding possible evaluation criteria were evaluated within the phases. Typical neck kinematics was seen for the two different situations. Occupants of lower weight had a more extended neck in the initial protraction phase and also a generally more pronounced upper neck link angle. Occupants with initial arm resistance had generally greater lower neck link angle at the time when the upper neck link angle was straight. No evaluation criteria reflected the anticipated AIS 1 neck injury rate consistently. In the initial protraction phase, NICmin correlated to expected injury outcome in almost half of the cases. In the protraction-flexion shift phase, Nkm, Nij, upper neck shear force and neck tension force reflected anticipated severity outcome to some extent. In the flexion phase, upper and lower neck extension correlated to anticipated AIS 1 neck injury rate only to a minor extent. The different sitting postures were more influential than the different crash pulses, emphasizing the importance of not only considering the spectra of impact severity but also differences in sitting postures in safety system development and evaluation. PMID- 15545078 TI - WalkSafe: a school-based pedestrian safety intervention program. AB - OBJECTIVE: To evaluate the effectiveness of the WalkSafe program, an elementary school-based pedestrian safety program in a single high-risk district in Miami Dade County. METHODS: Sixteen elementary schools were identified in a single high risk district and enrolled in a one-year study. All schools implemented the WalkSafe program on the last week of January 2003. A pre, post, and three-month post testing of pedestrian safety knowledge was conducted. An observational component was also implemented at four schools that were randomly chosen. Engineering recommendations and law enforcement initiatives were also performed. RESULTS: A total of 6467 children from K-5th grade from 16 elementary schools participated in the program. Of these 5762 tests were collected over three different testing times. A significant difference (p-value < 0.001) was found between the pre and post testing conditions across all grade levels. There was no significant difference found between the post and three-month post testing conditions across all grade levels (p-value > 0.05). The observational data collected at four schools across the different testing times demonstrated a significant difference found between pre and post testing conditions for Group A (stop at the curb and look left, right, left) and also for Group B (midstreet crossing and darting out) (p-value < 0.05). There was no difference found between comparing the pre-test or post-test condition with the three-month post-test time. There were many environmental modifications that were recommended and actually performed. CONCLUSION: The WalkSafe program implemented in a single high risk district was shown to improve the pedestrian safety knowledge of elementary school children. The observational data demonstrated improved crossing behaviors from pre-test to post-testing conditions. Future research will focus on sustaining the program in this district and expanding the program throughout our county. PMID- 15545079 TI - New biofidelic corridor and biofidelity test procedure for pedestrian legform impactors. AB - The objectives of this research are to propose a new impact response corridor for the ISO legform impactor and to determine the biofidelity of the current legform impactor with rigid leg and thigh developed by the Transport Research Laboratory (TRL). The latest data obtained from Post Mortem Human Subject (PMHS) knee impact tests were analyzed in connection with the proposal, and biofidelity legform impact tests were conducted using the current rigid legform impactor. New normalized biofidelic corridors of impact force corresponding to adult male 50th percentile (AM50) are proposed. The impact test results indicate the current rigid legform impactor does not have sufficient human knee biofidelity. The present results suggest that human tolerance can not be used directly for the injury reference value of the legform impactor. A conversion method is needed to interpret the data measured by current legform impactors as the injury reference value. PMID- 15545080 TI - In honour of Professor Rudy Juliano, winner of the 2004 Life-time Achievement Award from the Journal of Drug Targeting. PMID- 15545081 TI - A personal reflection on the career of the 2004 Life-time Achievement Award winner. PMID- 15545082 TI - The design and exogenous delivery of siRNA for post-transcriptional gene silencing. AB - RNA interference (RNAi) is a natural cellular process that effects post transcriptional gene silencing in eukaryotic systems. Small interfering RNA (siRNA) molecules are the key intermediaries in this process which when exogenously administered can inhibit or "silence" the expression of any given target gene. Thus, siRNA molecules hold great promise as biological tools and as potential therapeutic agents for targeted inhibition of disease-causing genes. However, key challenges to the effective and widespread use of these polyanionic, macromolecular duplexes of RNA are their appropriate design and efficient delivery to cells in vitro and in vivo. This review highlights the current strategies used in the design of effective siRNA molecules and also summarises the main strategies being considered for the exogenous delivery of siRNA for both in vitro and in vivo applications. PMID- 15545083 TI - Plasmid based gene delivery for orthopedic disorders: a brief review. PMID- 15545084 TI - Sulfhydryl based cationic surfactants and the impact of polyanions on disulfide bond formation: implications for gene transfer vectors. AB - Compacting plasmid DNA (pDNA) into a small size is a fundamental necessity for the efficient in vivo transfer of nucleic acids to somatic cells. An approach for accomplishing this is to condense pDNA using cationic detergents with sulfhydryl groups, near their critical micelle concentration. In this study, a model surfactant was used to study how the rate of disulfide bond formation relates to environmental factors. It was shown that the thiol detergent had the ability to form a disulfide bond when oxidized and the presence of polyanions was significantly increased. The addition of a reducing agent disrupted the disulfide bonds initially, but this was followed by disulfide bond reformation in a short time period. PMID- 15545085 TI - Novel long-circulating liposomes containing peptide library-lipid conjugates: synthesis and in vivo behavior. AB - Rapid uptake of intravenously injected liposomes by the mononuclear phagocyte system has limited their use as drug delivery vehicles. Recently, various long circulating liposomes have been prepared by incorporating glycolipids or other amphiphilic molecules into the lipid bilayer of conventional liposomes. The purpose of the present study was to design a new class of biodegradable membrane modifiers that would increase the half-life of liposomes in vivo. Using solid phase peptide synthesis, synthesized were 30-residue random libraries consisting of a random sequence of glycine, beta-alanine and gamma-aminobutyric acid. The libraries were coupled to stearic acid (SA) or phosphatidylethanolamine (PE). The resulting amphiphilic conjugates were mixed with egg phosphatidylcholine (PC) and cholesterol (Chol) in a 6:47:47 ratio, and unilamellar liposomes were prepared. For comparison, plain PC/Chol (50:50) liposomes, as well as liposomes containing polyethylene glycol (PEG)-SA/PC/Chol (6:47:47) and PEG-PE/PC/Chol (6:47:47) were also prepared. Calcein was entrapped in the liposomes, which were given intravenously to rats at a dose of 9.2 mumol lipid/kg, and the amount of intact liposomes present in serum was followed with time. While the conventional liposomes had a short elimination half-life (28 min), the liposomes modified with library-PE had a much longer half-life (170 min), while library-SA provided no improvement of the liposome pharmacokinetics. PEG-PE greatly improved the half life of the liposomes (400 min) while PEG-SA only provided a marginal improvement. All liposome preparations were cleared in a biphasic fashion. In conclusion, a novel biodegradable lipopeptide conjugate was designed that endows liposomes with a prolonged circulation time in vivo. The pharmacokinetic profile of these modified liposomes was drastically improved over that of conventional liposomes. Since the library is prepared by solid-phase synthesis, length and/or composition could easily be modified in order to modulate the clearance profile of the liposomes. Tailoring of the pharmacokinetic profile of the liposomes depending on their intended application may allow for a greater flexibility of use than PEG-PE. PMID- 15545086 TI - Sequence specificity of alternating hydroyprolyl/phosphono peptide nucleic acids against zebrafish embryo mRNAs. AB - Morpholino phosphorodiamidate (MO) DNA mimics display excellent water solubility and hybridization properties toward DNA and RNA, and have been utilized in the model vertebrate zebrafish (Danio rerio) for genome-wide, sequence-based, reverse genetic screens during embryonic development. Peptide nucleic acids (PNAs) exhibit excellent mismatch discrimination, nuclease resistance, and protease resistance, but low solubility. Negatively charged DNA mimics composed of alternating residues of trans-4-hydroxy-L-proline peptide nucleic acid monomers and phosphono peptide nucleic acid monomers (HypNA-pPNA) combine all of the positive features of both MOs and PNAs. Thus, we evaluated PNA oligomers and HypNA-pPNA oligomers as an alternative to MOs for oligonucleotide inhibition of gene expression in zebrafish embryos. We observed that HypNA-pPNA 18-mers displayed comparable potency to MO 25-mers as knockdown agents against chordin, notail and uroD, with greater mismatch stringency. Furthermore, we observed that a specific HypNA-pPNA 18-mer elicited the dharma (bozozok)(-/-) phenotype in zebrafish embryos, which MO 25-mers do not. These observations validate HypNA pPNAs as an alternative to MO oligomers for reverse genetic studies. The stronger hybridization and greater specificity of HypNA-pPNAs enable knockdown of mRNAs unaffected by MO oligomers. PMID- 15545087 TI - Polymer design and incorporation methods for polymeric micelle carrier system containing water-insoluble anti-cancer agent camptothecin. AB - A water-insoluble anti-cancer agent, camptothecin (CPT) was incorporated to a polymeric micelle carrier system forming from poly(ethylene glycol) poly(aspartate) block copolymers. Incorporation efficiency and stability were analyzed in correlation with chemical structures of the inner core-forming hydrophobic blocks as well as with incorporation methods. Among three incorporation methods (dialysis, emulsion and evaporation methods), an evaporation method brought about much higher CPT yields with less aggregation than the other two methods. By the evaporation method, CPT was incorporated to polymeric micelles in considerably high yields and with high stability using block copolymers possessing high contents of benzyl and methylnaphtyl ester groups as hydrophobic moieties. This indicates importance of molecular design of the hydrophobic block chain to obtain targeting using polymeric micelle carriers as well as importance of the drug incorporation method. PMID- 15545088 TI - Nutraceutics and delivery systems. AB - Medical treatment has been shifted to being more prophylactic as a recent trend. Postgenomic research has unveiled the fact that nutritional intervention has been strongly associated with genetic expressions, which are responsible for a variety of biological functions. Based on these findings, the prophylactic effects of dietary supplement and nutrient have been enthusiastically investigated. Preventing or retarding the onset of diseases has become a more attractive and cost effective strategy in the medical arena. Among other approaches to prevent diseases, antioxidants, which are found in many phytochemicals, have received much attention. However, most natural antioxidants such as alpha-tocopherol, ascorbic acid and others are biologically unstable, poorly soluble in water, and poorly distributed to target sites. Because of these shortcomings further prophylactic applications of dietary supplements have stagnated. This is partially due to a lack of basic awareness of drug delivery system for dietary supplements and nutrients. In this article, we strongly advocate serious consideration of the bioavailability of dietary supplements. Currently, there are some challenging works to improve their bioavailability using delivery systems such as liposomal formulations. We will discuss the target molecules of dietary supplements for prevention of diseases and also introduce the pioneering works of delivery systems for dietary supplements to promote their therapeutic value. PMID- 15545089 TI - Sustained polymeric delivery of gene silencing antisense ODNs, siRNA, DNAzymes and ribozymes: in vitro and in vivo studies. AB - Small interfering RNA (siRNA), antisense oligonucleotides (ODNs), ribozymes and DNAzymes have emerged as sequence-specific inhibitors of gene expression that may have therapeutic potential in the treatment of a wide range of diseases. Due to their rapid degradation in vivo, the efficacy of naked gene silencing nucleic acids is relatively short lived. The entrapment of these nucleic acids within biodegradable sustained-release delivery systems may improve their stability and reduce the doses required for efficacy. In this study, we have evaluated the potential in vitro and in vivo use of biodegradable poly (D,L-lactide-co glycolide) copolymer (PLGA) microspheres as sustained delivery devices for ODNs, ribozyme, siRNA and DNA enzymes. In addition, we investigated the release of ODN conjugates bearing 5'-end lipophilic groups. The in vitro sustained release profiles of microsphere-entrapped nucleic acids were dependent on variables such as the type of nucleic acid used, the nature of the lipophilic group, and whether the nucleic acid used was single or double stranded. For in vivo studies, whole body autoradiography was used to monitor the bio-distribution of either free tritium-labelled ODN or that entrapped within PLGA microspheres following subcutaneous administration in Balb-c mice. The majority of the radioactivity associated with free ODN was eliminated within 24 h whereas polymer-released ODN persisted in organs and at the site of administration even after seven days post administration. Polymer microsphere released ODN exhibited a similar tissue and cellular tropism to the free ODN. Micro-autoradiography analyses of the liver and kidneys showed similar bio-distribution for polymer-released and free ODNs with the majority of radioactivity being concentrated in the proximal convoluted tubules of the kidney and in the Kupffer cells of the liver. These findings suggest that biodegradable PLGA microspheres offer a method for improving the in vivo sustained delivery of gene silencing nucleic acids, and hence are worthy of further investigation as delivery systems for these macromolecules. PMID- 15545090 TI - Bonesetting and radiography in the southern Maya highlands. AB - Maya bonesetters and their clients have been affected by the entry of radiographic technology into highland Guatemala. Bonesetters rely on a form of embodied knowledge to perform their manual work, the kind of knowledge X-rays threaten to supersede. This article examines how Maya bonesetters are meeting this challenge to their legitimacy and how they are positioning themselves pragmatically within a world of changing biomedical resources and contested health knowledge. PMID- 15545091 TI - Breathless in houston: a political ecology of health approach to understanding environmental health concerns. AB - A political ecology approach to the study of environmental health problems can provide a comprehensive analytical framework with which to understand geographical and social disparities in health status. To date, however, political ecology has remained limited in its application to health problems, and where health has been addressed, biomedical models have prevailed, with little attention to differing explanatory models of health and disease. By integrating political ecology with an interpretive critical medical anthropology, one can better understand the ways in which health and environment intersect, and the differing social responses to environmental practices that affect human health. In this paper I summarize these theoretical issues and then discuss how this theory can be applied toward an analysis of air quality and health in Houston, Texas. This research suggests that local understandings of respiratory health often contradict public health concepts of environmental health and, in turn, differentially shape people's interactions with the environment. PMID- 15545093 TI - Slave ethics and imagining critically applied anthropology in public health research. AB - For several years I considered myself a resident enemy alien of public health research. This polemical article explores this enemy alien subjectivity by looking at an ethnographic case showing how race is figured in public health research, and it asks what this subjectivity might suggest for a medical anthropology struggling to be both more public and interdisciplinary, and more fundamentally ethnological. As I imbue my enemy alien subjectivity with uniquely anthropological images and references, I ask medical anthropologists to reflect upon the specialized nature of a contemporary anthropological imagination and, while doing this, to look to the Nietzschean notion of slave ethics in order to engage with questions of how we might use this self-awareness to create various modes of"postcritical"practice in public health. PMID- 15545095 TI - A 35-month profilometric and clinical evaluation of non-ablative remodeling using a 1540-nm Er:glass laser. AB - BACKGROUND/OBJECTIVE: As remodeling is getting more popular with patients, long term studies are becoming necessary. The aim of this 35-month clinical study was to evaluate the long-term benefits obtained using a 1540-nm Er:glass laser for non-ablative remodeling of perioral and periorbital rhytids. The role of maintenance treatments was also investigated. STUDY DESIGN/METHODS: Eleven women with periorbital and perioral rhytids underwent a series of five treatments at 6 week intervals with an Er:glass laser. Five patients subsequently received two maintenance retreatments and six did not. The maintenance treatments were performed at 14 and 20 months. Silicone imprints were performed to measure anisotropy before treatment, at 6 months, at 14 months and at 35 months. Patient self-evaluation/questionnaire was also done to assess adverse effects and subjective clinical improvement. RESULTS: For all 11 patients, the percentage of anisotropy reduction was 41.21% at 6 months, 51.76% at 14 months and 29.87% at 35 months. No adverse effects were noted. Patient satisfaction was high at the end of the evaluation. Retreated patients were more satisfied than non-retreated ones. However, there was no difference in the anisotropy factor between the two groups. CONCLUSION: Treatment of facial rhytids with a non-ablative 1540-nm Er:glass laser system can produce benefits that persist over 2 years after the last treatment. PMID- 15545096 TI - Long pulsed dye laser treatment of facial wrinkles. AB - BACKGROUND: The flashlamp pulsed dye laser has been used in the treatment of rhytids. OBJECTIVE: To evaluate the efficacy of the long pulsed dye laser in the treatment of mild to moderate wrinkles in Asian patients. METHODS: Wrinkles on one half of the face in 10 subjects were treated with the long pulsed dye laser (595 nm, 10 mm spot size, 10 ms, 7 J/cm2, 40 ms spray, 40 ms delay, single-pass, 30% overlap) with the other side serving as a control. A total of three treatments were given at 2 monthly intervals. The following sites were treated: periorbital area, six patients; forehead, two patients; cheek, two patients. No preoperative anesthesia or postoperative treatment were used. Clinical photographs were taken before and after each treatment, and analysis was undertaken through photographic evaluation by non-treating physicians. RESULTS: At 2 months after the last treatment, the clinical improvement of rhytids was noted in all patients compared with baseline. Four subjects had mild improvement (< or = 25%), five had moderate improvement (26-50%) and one had marked improvement (51-75%). The periorbital area was more responsive to treatment compared with the other sites. No clinical changes were noted in the control areas. No adverse effects were reported except for transient mild erythema in two patients which lasted for up to an hour. Nine patients were somewhat satisfied with the treatment and one was highly satisfied. All wanted the other half of the face to be treated. CONCLUSION: Treatment with a non-ablative 595 nm flashlamp pulsed dye laser can lead to mild to moderate clinical improvement in class I-II rhytids with minimal to no side effects in patients with darker skin types. PMID- 15545097 TI - Clinical evaluation of non-ablative radiofrequency facial rejuvenation. AB - OBJECTIVE: To evaluate the clinical efficacy of non-ablative cutaneous radiofrequency (RF) facial rejuvenation. DESIGN: Prospective study with longitudinal follow-up of a validation cohort set in an urban, private practice of an ambulatory facial plastic surgery center in southern California. METHODS: A consecutive sample was enrolled of 35 healthy adults with moderate facial aging, manifested by skin laxity, rhytids, and ptosis (brow, midface, jowls). Following intravenous sedation anesthesia, the study area was treated with 115-144 J/cm2 using the non-ablative RF device (ThermaCool TC). The main outcome measures were the objective measurement of brow height, investigator evaluation of skin parameters, a patient satisfaction questionnaire, and standardized photography, at fixed time intervals. RESULTS: At 12 weeks, a statistically significant increase in mean vertical brow height of 1.6-2.4 mm was observed in patients treated exclusively with the RF device (p<0.0001). All skin parameters (laxity, wrinkles, clarity, pore size) were improved. Complications and side effects were minimal. Patients were uniformly satisfied. CONCLUSIONS: The ThermaCool TC RF system represents a promising non-invasive method of obtaining moderate facial rejuvenation in the appropriately selected patient. Long-term results are pending. PMID- 15545098 TI - Botulinum toxin type A gives adjunctive benefit to periorbital laser resurfacing. AB - OBJECTIVE: Periorbital aging and lines are a result of intrinsic skin aging, ultraviolet damage, and repetitive action of periorbital muscles. Rejuvenation of this area should therefore be optimized by combining treatments that approach the different causative factors. METHODS: This was a single-center, prospective, randomized, placebo-controlled study comparing the efficacy and safety of combining Botox injections (18 units per area) with ablative laser resurfacing versus laser resurfacing alone without Botox in the treatment of periorbital rhytids. RESULTS: We have concluded a bilateral study comparing the effects of Botox versus saline placebo injections to the periorbital areas before and following erbium: YAG laser resurfacing of the areas in 33 patients. The results demonstrated that the Botox-treated side with laser resurfacing improved significantly more than the contralateral area treated with saline and laser in diminishing periorbital rhytids as well as textural, pigmentation, and other features of periorbital skin aging. CONCLUSION: This study illustrates the benefits of a combined approach to treating periorbital skin aging. PMID- 15545099 TI - Resolution of retracted scar after 585-nm pulse dye laser surgery. AB - BACKGROUND: Laser scar revision has been an effective method for improving several aspects of scarring through ablative and non-ablative sources. The 585-nm pulsed dye laser (PDL) is an important non-ablative instrument for reducing scar bulk and symptoms. OBJECTIVE: To describe the use of a 585-nm PDL for the treatment of a retracted and atrophic facial scar. METHODS: We report the case of a 26-year-old patient who presented with a retracted facial scar following surgical excision of an aggressive benign tumor. Treatment was carried out using the 585-nm PDL. RESULTS: Treatment of the scar using two low-level PDL therapies significantly altered the appearance of the scar and augmentation of the retracted defect was avoided. CONCLUSION: Treatment of this retracted and atrophic facial scar with the 585-nm PDL was very effective and safe. PMID- 15545100 TI - A prototype vessel compressor helps efficient laser treatment of small leg veins. AB - The eradication of small leg veins with lasers continue to present problems. Visible light lasers (488 nm approximately 595 nm) are well absorbed in haemoglobin but melanin is also a target, necessitating aggressive skin cooling to prevent damage to the epidermis and adding to the expense of these laser systems. A new generation of much less expensive semiconductor-based lasers operating in the near infrared offers a different approach, with protein as the main target rather than pigment. For visible light lasers, compression of the target vessels is a contraindication, since the target pigment is removed. For near IR diode lasers, however, compression of the vessels is a benefit, as the cooling effect of the blood flow is removed and the highly proteinous vessel walls are coapted which encourages efficient coagulative vessel closure. The prototype of a simple vessel compressor is presented, which first compresses vessels and coapts the walls, and then presents the coapted vessels as a target for a laser of an appropriate wavelength. PMID- 15545101 TI - 420 nm intense continuous light therapy for acne. AB - BACKGROUND: Topical antibiotics, isotretinoin or systemic antibiotics are usually used for acne therapy. However, isotretinoin cannot be used during pregnancy because it can cause significant birth defects while systemic antibiotics can have adverse side effects such as gastrointestinal irritation, photosensitivity and tetracycline sensitivity. Describe here is a high-intensity, narrow-band, blue light (ClearLight) system, and its therapeutic clinical effect is investigated on acne using cutaneous measurements, bacterial observations and ultrastructural changes. MATERIALS AND METHODS: A total of 28 adult healthy volunteers with facial acne (mean age 28.1 years, range 16-56 years) were recruited for this study. They were treated with a total of eight serial biweekly 15-minute treatment sessions. Clinical counts of acne, as well as moisture, sebum and pH measurements were taken between each session. Nine of the 28 patients were followed for 2-3 months after the last treatment. Detection of bacteria in acne pustules was analyzed by culture and by polymerase chain reaction (PCR). Ultrastructural changes were examined in eight patients after four sessions of the light therapy. RESULTS: All patients completed the study. Overall, there was a 64.7% improvement in acne lesions. There were no bacterial changes before or after the therapy, although damaged Propionibacterium acnes were observed at the ultrastructural level. CONCLUSIONS: ClearLight performed eight times over 4 weeks can be useful in the treatment of acne. Further investigation will be needed to elucidate the mechanism of action of ClearLight. PMID- 15545102 TI - Smoothbeam laser treatment may help improve hidradenitis suppurativa but not Hailey-Hailey disease. AB - OBJECTIVE: The attributes of the Smoothbeam laser--a 1450 nm diode laser--were considered likely to improve two distinct chronic conditions of flexural skin: Hailey-Hailey disease (HHD) and hidradenitis suppurativa (HS). METHODS: A series of laser treatments were given to the axillae of one patient with each condition. RESULTS: Treatments were painful but tolerated. There were no adverse sequelae following laser treatment. For the patient with HS, partial improvement was achieved after four treatments and sweating was notably reduced. Intermittent laser treatment may be needed to maintain or build upon the improvements gained. Longstanding HS cases, however, with established sinuses and thick scar tissue are unlikely to be helped by this form of laser treatment. There was no subjective or objective improvement seen in the patient with HHD. The only notable beneficial effect was a marked reduction in sweating and elimination of the malodour. CONCLUSIONS: Subsurfacing laser treatment in contrast with resurfacing laser treatment has no benefit in the management of HHD. Why the Smoothbeam laser causes sweat reduction is not known--and its clinical value has yet to be established. PMID- 15545103 TI - Localisation and William Macewen's early brain surgery Part I: the controversy. AB - Neurosurgery for the removal of brain tumours based on localising signs is usually dated from the 1884 operation by Bennett and Godlee. However, within weeks of that operation claims were made on behalf of William Macewen, the Glasgow surgeon, to have been the real pioneer of such surgery. According to Macewen's protagonists, he had conducted seven similar operations earlier than Bennett and Godlee and, in a notable 1888 address, Macewen described these seven pre-1884 cases and a number of others operated on after 1884. This paper, which is in two parts, contains an evaluation of the claims made for the priority of Macewen's pre-1884 operations. Part I deals mainly with Macewen's work in fields other than brain surgery that are relevant to it and sets out the facts of the controversy. It begins with a brief biography of Macewen, describes his pioneering work in antiseptic and aseptic surgery, his work on osteotomy and bone regeneration, and his use in brain surgery of the knowledge so gained. Part I concludes with an examination of the battle waged in the newspapers between Macewen's and Bennett's and Godlee's supporters, and of previously unpublished correspondence between Macewen himself, David Ferrier and Hughes Bennett. The primary records of the patients on whom Macewen operated, together with other materials relevant to the controversy, are examined in Part II. PMID- 15545104 TI - Mitchell's influence on European studies of peripheral nerve injuries during World War I. AB - OBJECTIVE: Describe the influence of S. Weir Mitchell's (1829-1914) work, and in particular his ideas on causalgia, on European physicians who treated peripheral nerve injuries during World War I (WWI). BACKGROUND: During the American Civil War (1861-1865), Mitchell studied peripheral nerve injuries with colleagues George Read Morehouse and William Williams Keen. Three monographs resulted from this work. All were important landmarks in the evolution of knowledge of peripheral nerve injuries. A subsequent occasion to improve knowledge came in WWI. METHODS: The most important European monographs or series on peripheral nerve injuries from WWI were studied with special interest in references to causalgia and Mitchell's works on peripheral nerve injuries. We included works by Tinel, Athanassio-Benisty, Purves-Stewart & Evans and Carter, Foerster and Oppenheim. RESULTS: Tinel and Athanassio-Benisty provided the most detailed information on peripheral nerve injuries and causalgia and often referred to Mitchell. Both mentioned a possible sympathetic origin. Athanassio-Benisty described tremor and other movement disorders in relation to causalgia. Purves Stewart and Evans mentioned Mitchell and causalgia in the second edition of their book. They advocated the term "thermalgia." Carter, who had access to data of many cases, concentrated his work on causalgia, referring to Mitchell. Foerster provided data of a great number of peripheral nerve injuries, but did not refer to Mitchell. However, he described the symptoms of causalgia cursorily, applying the term Reflexschmerz (reflexpain). Oppenheim was particularly interested in muscle innervation and referred to Mitchell with respect to hypertrichosis and glossy skin. Oppenheim did not use the term causalgia, although he described the syndrome in some of his patients. It wasn't until around 1920 that German physicians devoted significant attention to causalgia and began using the term. CONCLUSION: Knowledge of peripheral nerve injuries was greatly advanced during and after WWI. Mitchell's influence was mainly found in the French medical literature, where his findings provided the basis for further research on the origin of causalgia. In England, Mitchell and causalgia were also well-known. We found evidence to suggest that some of the English knowledge came from French physicians. German physicians described the symptoms of causalgia, but did not use the term, nor did they refer to Mitchell. This variation in Mitchell's influence by country probably reflects the fact that Mitchell's Injuries of nerves and their consequences was translated into French but not German. PMID- 15545105 TI - Illnesses of the brain in John Quincy Adams. AB - John Quincy Adams, the sixth and perhaps most scholarly American president, served courageously despite familial essential tremor, depression, and cerebrovascular disease. His cousin Samuel Adams and his father John Adams also had essential tremor, which the later called "quiveration". Alcoholism and depression affected several members of J.Q. Adams's family. Following his own time as president, J.Q. Adams returned to duty as the congressman who most assiduously fought slavery, a fight he continued even after he had suffered a major left hemispheric stroke. His fatal collapse in Congress, protesting the Mexican War, is legendary among the final illnesses of American statesmen. PMID- 15545107 TI - The history of the (re)naming of multiple sclerosis. PMID- 15545106 TI - A brief exploration of neurological art history. AB - The invention of realistic portraiture to reveal "inner life" is attributed by some art historians to Jan van Eyck who worked in Flanders from 1420 onwards. We show, using clinical neurological examination of the gold mask of Agamemnon dating from 1550-1500 BC and of the portraits of Henry III and his son Edward I - important English royals -- painted between 1216 and 1307, that realistic portraits were made well before the 15th Century. Thus artists unwittingly used neurology as part of their realistic approach to the presentation of the face. Because neurological diagnosis is often visual, neurology, in turn, has a rich potential to unveil examples of realism in art. We consider the art pieces examined here also pertinent to art historians, as they assess the role of art in documenting history. PMID- 15545108 TI - The Swiss advisor. PMID- 15545109 TI - Francis Schiller (1909-2003). PMID- 15545110 TI - TENNET - ISHN Joint Conference. Ninth Annual Meeting of the International Society for the History of the Neurosciences. June 26-29, 2004. PMID- 15545113 TI - Neither Convention nor Constitution--what the debate on stem cell research tells us about the status of the common European ethics. PMID- 15545114 TI - How traditions of ethical reasoning and institutional processes shape stem cell research in Britain. AB - This article aims to show how the traditions of ethical reasoning and policy making shape stem cell research in Britain. To do so I give a detailed account of the earlier developments of regulations on embryo research and the specific scientific advances made in Britain. The subsequent regulation of stem cell research was largely predetermined by those structures and the different and partly opposing orientations of a utilitarian approach to policies on biomedicine. The setting up of the first stem cell bank and the directing of public funding into not only bioethical but also sociological guidance of the development of the new science field are aspects of the particular British way of supporting stem cell research. However, there is also an ongoing philosophical and juridical debate on the possible erosion of fundamental values caused by incremental regulatory weakening. Although I am highly sympathetic to the critical position that there is a need for a metaphysical anchor to secure individual human rights, one has to admit that the British mode of handling the inevitable ethical problems we face with biomedical progress is rather successful in terms of securing some of the basic needs and values of a modern democratic society. PMID- 15545115 TI - The ethics of embryonic stem cell research. AB - In this article I rebut conservative objections to five phases of embryonic stem cell research. I argue that researchers using existing embryonic stem cell lines are not complicit in the past destruction of embryos because beneficiaries of immoral acts are not necessary morally tainted. Second, such researchers do not encourage the destruction of additional embryos because fertility clinics presently destroy more spare embryos than researchers need. Third, actually harvesting stem cells from slated-to-be-discarded embryos is not wrong. The embryos are not sacrificed for the good of others because they would have been destroyed anyway. Fourth, harvesting stem cells from embryos that are not doomed is morally acceptable, because preserving frozen embryos is futile therapy. Finally, creating embryos solely for the sake of harvesting stem cells from them is morally acceptable because the assumption that embryos have the right to life has very counterintuitive implications. PMID- 15545116 TI - The European embryonic stem-cell debate and the difficulties of embryological Kantianism. AB - As elsewhere, the ethical debate on embryonic stem cell research in Central Europe, especially Germany and Switzerland, involves controversy over the status of the human embryo. There is a distinctive Kantian flavor to the standard arguments however, and we show how they often embody a set of misunderstandings and argumentative shortcuts we term"embryological Kantianism."We also undertake a broader analysis of three arguments typically presented in this debate, especially in official position papers, namely the identity, continuity, and potentiality arguments. It turns out that these arguments do not support the strong, quasi-personal status accorded to the embryos in these official opinions. PMID- 15545117 TI - The embryo in relationships: a French debate on stem cell research. AB - While many European countries are entering unknown legal terrain where the embryo in vitro is concerned, France can already look back on a long tradition of public discussion and legal codification of ways of dealing with in vitro embryos. In its comprehensive law of 1994, France had still rejected embryo research; however, due to the promising perspectives of stem cell research, the new law now pending implies a clear liberalization of the 1994 provisions. Both the French lawmakers and the National Ethics Commission have repeatedly argued that possible utilization of embryos for research purposes may seem legitimate from the moment that there is no more "parental project." De facto, this concept implies that an embryo can be transformed into an object from the moment that the parents cease to desire it and that the value of protection is solely dependent on the will of third persons. At the same time, France is still speaking of guaranteeing respect for the "dignity of the embryo," which would mean that an embryo must not be reduced to a thing and treated for purposes which are not his own. Therefore, the French solution is not a consistent and honest solution, and in its new legal provisions, France has involved herself in manifold contradictions. France has rejected the conception of pre-embryo, but is de facto following Britain's model without making it explicit. PMID- 15545118 TI - On the German debate on human embryonic stem cell research. AB - Germany since 1990 has one of the strictest human embryo protection laws, yet according to the Stem Cell Act of 2002 allows, under strict conditions, the import and use of human embryonic stem cells (hESC) for high priority research goals. The author tries to show how this is taken to be coherent by the parliamentary majority (though not necessarily by the general public) in Germany. In doing so, he firstly looks into the chronicle of the debate in Germany showing its different stages since 1999, then dwells upon the relation between the law and the role of ethics in this issue, and thirdly presents the two fundamentally different positions of the German debate, that is, that the human embryo created for IVF purposes is a human being and stands from its very beginnings under the constitutional principles of respect for, and protection of, human life versus the position that before being implanted the human embryo may become a human being and therefore belongs to the human species only potentially, so that its right to life protection may be assessable over against other high priority goals, such as research aiming at possible help for patients with life endangering diseases. In spite of the Stem Cell Act of 2002, the debate of the German general public goes on, especially due to the recent EU 6th Research Framework Program which plans to also fund hESC research. PMID- 15545119 TI - Preimplantation genetic diagnosis (PGD): European perspectives and the German situation. AB - This article gives an overview about the ethical dispute on preimplantation genetic diagnosis (PGD), its legal status and its practical usage in Europe. We provide a detailed description of the situation in Germany wherein prenatal diagnosis is routinely applied, but PGD is prohibited on the basis of the internationally unique embryo protection act (EPA) that was put into force in 1991. Both PGD and stem cell research were vigorously debated in Germany during the last four years. As regards the PGD debate specifically, the voices of the ones directly affected were not adequately taken into consideration. We describe the predominant lines of argumentation in this debate and some essential results of our "bioethical field study" of opinions on and usage of PGD in Germany and their implications for the German legislation and ethical theory. PMID- 15545120 TI - Patient satisfaction in our initial experience with endonasal endoscopic non laser dacryocystorhinostomy. AB - BACKGROUND/AIMS: Endonasal endoscopic dacryocystorhinostomy (EN-DCR) is now a well-established procedure to relieve nasolacrimal duct obstruction. In the past, attempts have been made to comment on the anatomical success of the procedure. However, no studies have been conducted to evaluate patient satisfaction with the EN non-laser DCR procedure in comparison with the surgeon's experience. METHODS: Records of patients undergoing EN-DCR and external DCR (ET-DCR) surgery were reviewed. A telephone questionnaire was used to assess patient satisfaction with both procedures. Data were analyzed with Fisher's exact test and the chi-square test. RESULTS: Twenty primary EN-DCR's and 16 revision EN-DCR's were performed by the same surgeon (RM) over a three-year period. At last review, 89% of ET-DCR and 75% of EN-DCR procedures were noted to have a patent sac washout performed in the eye clinic. A telephonic interview revealed no significant difference between the surgical outcome [15/20 (75%)] and patient satisfaction [14/20 (70%)] with the primary EN-DCR procedure. Patient satisfaction with revision EN-DCR [10/16 (63%)] was slightly poorer than the surgical outcome recorded for revision EN-DCR [12/16 (75%)] but this was also not statistically significant. Telephonic interview was possible for 42/64 (66%) patients undergoing primary external ET-DCR's and a total of 36/42 (86%) patients were satisfied with the procedure. Patient age, laterality, duration of symptoms, previous ocular procedures or preexisting ocular disease and associated ENT procedures did not alter the surgical result or patient satisfaction in either group. CONCLUSIONS: This study suggests that patient satisfaction with endoscopic endonasal non-laser DCR for primary or revision DCR surgery is comparable to that with the external-DCR technique since there was no significant difference in patient satisfaction between the two groups of patients. Patient perception of their symptomatic improvement was lower (though not statistically significant) in relation to the final clinical assessment of the outcome of both primary and revision EN-DCR. PMID- 15545121 TI - Punch biopsy in the management of periocular basal cell carcinomas. AB - AIM: The aim of this study was to define the role of punch biopsy in the management of periocular basal cell carcinomas. PATIENTS AND METHODS: A case-note review was performed on 24 patients undergoing punch biopsies for eyelid lesions. Depending on the clinical diagnosis, two groups were identified: Group A with periocular basal cell carcinomas and Group B with periocular lesions other than basal cell carcinoma. The histopathological diagnosis based on punch biopsy specimens was compared with (a) the clinical diagnosis and (b) the final histopathological diagnosis based on the excision biopsy specimen. RESULTS: The punch biopsy confirmed the diagnosis of basal cell carcinoma in 14 of 16 cases in Group A and 2 of 8 cases in Group B. Clinical characteristics when used alone in the diagnosis of basal cell carcinoma had a positive predictive value of 87.5% and a negative predictive value of 75%; hence a sensitivity of 87.5% and a specificity of 75%. Appropriate surgical intervention (two-stage tumour excision and lid reconstruction) was performed in 2 of 8 cases (25%) not initially clinically diagnosed as basal cell carcinomas. Unnecessary surgical intervention was avoided in 2 of 16 cases (12.5%) clinically diagnosed as basal cell carcinomas. The final histopathological result in all cases undergoing surgical excision confirmed the diagnosis of basal cell carcinoma made from the punch biopsy tissue specimens. CONCLUSION: The results suggest that punch biopsy provides an accurate histopathological diagnosis prior to commencing definitive surgical management and may be a useful tool in the management of periocular basal cell carcinomas. PMID- 15545122 TI - Adult onset Langerhans cell histiocytosis of the orbit--a case report. AB - PURPOSE: To report an interesting case of adult onset Langerhans cell histiocytosis in a 41-year-old male with clinicopathological correlation. DESIGN: Interventional case report. MATERIALS AND METHODS: A 41-year-old male presented with an ill-defined mass occupying the temporal quadrant of the left orbit. Computerized tomography revealed an ill-defined extraconal mass involving the superolateral aspect of the left orbit with areas of osteolysis involving the lateral and superior orbital walls. The mass excised at orbitotomy showed microscopic features consistent with Langerhans cell histiocytosis. Immunohistochemistry with CD-68 macrophage marker and S-100 was positive, confirming the diagnosis. Treatment included oral steroids and radiotherapy. At 6 months follow-up, he developed a punched-out lytic lesion in the left parietal calvarium. He again received external beam radiotherapy. At 14 months follow-up, he is doing well with no recurrences. CONCLUSIONS: Adult onset Langerhans cell histiocytosis, though rare, should be included in the differential diagnosis of lytic lesions of the lateral orbital wall in an adult patient. PMID- 15545123 TI - Agger nasi cell mucosal autograft for lacrimal sac reconstruction during endonasal dacryocystorhinostomy. AB - A mucosal autograft composed of agger nasi cell mucosa was used to reconstruct the lacrimal sac mucosa in a young woman with a cicatrised lacrimal sac and obstructed nasolacrimal system. The patient had undergone a previous external dacryocystorhinostomy (DCR), dacryocystectomy and revision mechanical endonasal DCR. There was persistent epiphora and at further mechanical endonasal (MEN) DCR it was noted that only a small cuff of lacrimal sac mucosa around the common canaliculus was present. To achieve a mucosal anastomosis, the agger nasi cell mucosa was removed from the cell and placed as a free mucosal graft around the common canalicular opening. Apposition with the nasal mucosa was completed with flaps from the lateral nasal wall mucosa. Postoperatively, the patient is asymptomatic with a freely draining ostium at 18 months of follow-up. PMID- 15545124 TI - Mobile orbital Cysticercus cyst--an unusual presentation. PMID- 15545125 TI - Orbital lymphoma mimicking optic nerve meningioma. AB - PURPOSE: To describe two patients with orbital lymphoma mimicking optic nerve meningioma. METHODS: Retrospective case review. RESULTS: Both patients presented in their sixth decade with gradually increasing proptosis and decreasing vision. Computerized tomography revealed homogeneous masses encasing the optic nerves, which in conjunction with the clinical presentations were felt to represent optic nerve meningiomas. Biopsy, however, led to a diagnosis of lymphoma of mucosal associated lymphoid tissue origin in one and a well-differentiated non-Hodgkin's lymphoma in the other. CONCLUSION: Orbital lymphoma surrounding the optic nerve may mimic the clinical and radiological appearance of an optic nerve meningioma. PMID- 15545127 TI - Normal exophthalmometry values: the need for calibrated exophthalmometers. PMID- 15545128 TI - Advancement of Whitnall's ligament via the conjunctival approach for correction of congenital ptosis. AB - PURPOSE: To conduct a preliminary study on ptotic eyelids with unilateral, congenital, simple partial ptosis, operated by advancement of Whitnall's ligament via the conjunctival route. METHOD: A prospective, interventional, clinical study. RESULTS: A total of 10 patients were recruited. The ptosis was 4 mm or less (range 2-4 mm; mean 2.7) and LPS action was greater than 5 mm (range 6-14 mm; mean 9.7). The ptosis was corrected by advancement of Whitnall's ligament by plication of the levator aponeurosis via the conjunctival route. Postoperatively, correction within 1 mm of the normal lid height was taken as good correction. All patients were assessed for a minimum of one year with regard to the amount of correction, residual ptosis, lid lag and lagophthalmos. In our study, good correction, i.e. within 1 mm of the normal fellow eye in primary gaze, was obtained in 80% of patients. Patients with more than or equal to 8 mm levator function gave better results. Undercorrection occurred in two patients. Lid lag and lagophthalmos occurred in all patients but was within acceptable limits. CONCLUSION: Advancement of Whitnall's ligament by plication of the levator aponeurosis via the conjunctival route is a viable procedure, which has all the advantages of performing the same technique via the cutaneous approach, i.e. shorter surgical time, minimal dissection of tissue planes and reversibility. The conjunctival approach has the added advantage of being cosmetically more acceptable as there is no visible scar on the lid. The technique also offers satisfactory functional results with few (correctable) complications. PMID- 15545129 TI - Reliability of exophthalmos measurement and the exophthalmometry value distribution in a healthy Dutch population and in Graves' patients. An exploratory study. AB - AIM: The purpose of this study was to test the reliability of an exophthalmometer commonly used in the Netherlands; to determine the exophthalmometry value distribution with this instrument and to assess the upper exophthalmometry limits of normal in a healthy, adult, Caucasian, Dutch population. Furthermore, to assess the effects of gender and age on exophthalmometry readings in this group and in a group of Graves' patients by comparing healthy, adult, Caucasian, Dutch individuals with adult, Caucasian, Dutch Graves' patients. METHODS: To test the reliability of our Hertel exophthalmometer, we determined the interobserver variation between two observers by measuring 160 eyes in healthy, adult, Caucasian, Dutch females and males (10 females and 10 males in each decade between 20 and 60 years of age). These data were also used for the assessment of the Hertel value distribution and for defining the upper limits of normal in these individuals by logistic regression analysis. The effects of disease, age and gender were established using these data plus data of a retrospective study of 393 adult, Caucasian, Dutch females (n=294) and males (n=99) with Graves' orbitopathy in whom Hertel values were measured with the same exophthalmometer. RESULTS: Exophthalmometry using an Hertel exophthalmometer appeared reliable (Pearson correlation coefficient for interobserver variation 0.89; 96% of the Hertel values measured by two observers were within the limits (of 2 mm) of agreement). Hertel values usually show a normal distribution in healthy individuals and in Graves' patients and are sex- and age-dependent, but there was no dependence on age in this small series in adults. Logistic regression analysis revealed an upper limit of normal of 16 mm in females and 20 mm in males in our group, using the exophthalmometer described. CONCLUSIONS: Exophthalmometry is reliable and absolute measurement of proptosis is feasible. International standardization of Hertel exophthalmometry is required in order to compare exophthalmometry data in the literature reliably. PMID- 15545130 TI - Dacryocystectomy: indications and results. AB - BACKGROUND: After the advent of dacryocystorhinostomy (DCR), dacryocystectomy (DCT) was regarded as mutilant surgery and reserved for lacrimal sac tumors. PURPOSE: To present current indications for dacryocystectomy. METHODS: Eighteen eyes from 11 patients subjected to DCT were reviewed retrospectively. Nine patients had chronic dacryocystitis confirmed by dacryocystography and two patients had lacrimal sac tumors. RESULTS: The indications for DCT were: three patients with systemic medical problems, three patients with dry eye, two cases of lacrimal sac tumors, two cases of traumatic dacryocystitis and one case of cutaneous leishmaniasis. Dacryocystitis was resolved in 17 of 18 eyes; the two cases of tumor evolved without epiphora. One patient (traumatic dacryocystitis) had recurrence of dacryocystitis. CONCLUSIONS: DCT is mainly performed when a lacrimal sac tumor is suspected but, since it avoids the intra- and postoperative complications related to dacryocystorhinostomy, it can be indicated in cases of dacryocystitis with significant lacrimal discharge and an enlarged or altered lacrimal sac. PMID- 15545131 TI - Dermatography as a treatment after periocular surgery. AB - Dermatography is the application of tattooing techniques in medicine. In the past, several techniques have been tried but none of these led to reproducible results. Over the last 17 years, dermatography has been developed into a technique that is both generally applicable and reproducible. It is a suitable treatment modality for disturbing skin discolorations and scars. With dermatography these can be permanently camouflaged. In this paper, the use of dermatography as a treatment after periocular interventions is discussed. PMID- 15545132 TI - Malignant peripheral nerve sheath tumour metastatic to the orbit. AB - PURPOSE: To present a case of malignant peripheral nerve sheath tumour (MPNST) metastatic to the orbit. METHOD/RESULTS: The authors describe a 29-year-old woman with disseminated MPNST who presented with acute right orbital inflammation. Computerized tomography revealed a superolateral mass with a central radiolucent area. At biopsy, the mass was primarily subperiosteal and histology showed a spindle cell, S100-positive MPNST consistent with the primary tumour. CONCLUSION: This is the second reported case of MPNST metastatic to the orbit and illustrates that an inflammatory presentation is possible in this aggressive malignancy. PMID- 15545133 TI - Nasal polyps herniating through lacrimal-nasal bony ostium mimicking dacryocystocele: a rare cause of DCR failure. AB - PURPOSE: To describe a rare cause of dacryocystorhinostomy (DCR) failure resulting from polyps herniating through the lacrimal-nasal ostium in a patient with severe, recurrent nasal polyposis. DESIGN/METHODS: Observational case report. Clinical practice setting. RESULTS: Five years after DCR surgery, a 65 year-old male patient presented with epiphora and a soft cystic enlargement in the lacrimal sac area mimicking a dacryocystocele. The Jones 1 test was negative but the Jones 2 test positive. Computer tomography (CT) showed bilateral obstruction of the nasal space by soft tissue, herniating through the lacrimal nasal ostium, suggestive of recurrent nasal polyposis. Transnasal polypectomy was performed, which re-established tear-drainage to the nose. CONCLUSIONS: Recurrent nasal polyposis may lead to DCR failure and may give rise to a soft tissue swelling at the site of previous DCR surgery. PMID- 15545134 TI - Orbital haemorrhage with loss of vision in a patient with disseminated intravascular coagulation and prostatic carcinoma. AB - A 65-year-old man with sudden profound loss of vision in his right eye due to sub periosteal orbital haemorrhage was found to have disseminated intravascular coagulation (DIC) secondary to metastatic prostatic carcinoma. CT-scan did not reveal any orbital metastases. A lateral canthotomy did not help to restore the vision. Orbital haemorrhage is known to occur with DIC due to different causes. To the best of our knowledge this is the first report of orbital haemorrhage with DIC related to prostatic carcinoma. This case emphasises the importance of considering systemic factors in cases of non-traumatic haemorrhage, along with imaging studies to rule out any co-existing vascular anomaly. PMID- 15545138 TI - Sinuorbital complications after intranasal cocaine abuse. AB - Proptosis and double vision were the presenting signs in a case of chronic orbital inflammation generated by osteolytic destruction of the sinuorbital barriers due to intranasal abuse of cocaine. The pathophysiologic background and reports from the literature dealing with orbital involvement in this condition are discussed. PMID- 15545139 TI - Factors influencing the detection of visual developmental deficits in 3-year-old kindergarten children. AB - PURPOSE/BACKGROUND: In the 'Tubingen Kindergarten study', an orthoptic screening program for amblyopia in three-year-olds was evaluated. In the retrospective analysis presented here, the association between a child's history--especially participation in the regular preventive care examinations in Germany (U4 to U7)- and the time of detection of target diseases was investigated. METHODS AND SUBJECTS: The parents of three groups of children were interviewed by telephone. Group 1 consisted of 21 children already treated for amblyopia before orthoptic screening; group 2 of 26 children with target conditions newly detected by orthoptic screening; and group 3 of 32 children with newly detected moderate ametropia. The groups were tested for differences in history and utilization/results of the regular preventive care examinations. RESULTS: Participation in regular preventive care ranged from 82% to 92%. There were no significant differences between the groups. Nearly 90% of parents from groups 2 and 3 had not heard of amblyopia before orthoptic screening. Only one child had been referred after regular preventive care examinations. CONCLUSIONS: The regular preventive care examinations contributed little to the detection of amblyopia. The strong position of the pediatrician should be used for the early information of parents. PMID- 15545140 TI - Anterior transposition compared to graded recession of the inferior oblique muscle for V-pattern strabismus. AB - OBJECTIVE: To compare the efficacy of anterior transposition (AT) and graded recession (GR) in the treatment of V- pattern strabismus caused by overaction of the inferior oblique muscle. METHODS: The charts of surgically treated cases of V pattern strabismus were analyzed retrospectively and the patients classified as AT or GR. Age, initial horizontal deviation, initial V-pattern and the amount of V-pattern correction were compared between the two groups. RESULTS: There was no significant difference in age (p = 0.066), initial horizontal deviation (p = 0.59), initial V-pattern (p = 0.15) or the amount of V-pattern correction (p = 0.78) between the two groups. CONCLUSION: AT is at least as effective as GR in the treatment of V-pattern strabismus caused by overaction of the inferior oblique muscle. PMID- 15545141 TI - Non-cycloplegic refractive screening can identify infants whose visual outcome at 4 years is improved by spectacle correction. AB - The Second Cambridge Population Infant Vision Screening Programme using the VPR-1 videorefractor without cycloplegia was undertaken in order to identify those infants with refractive errors who were potentially amblyogenic or strabismogenic. Infants identified at eight months were entered into a control trial of treatment with partial spectacle correction and underwent a long-term follow-up that monitored a wide range of visual, visuoperceptual, visuocognitive, visuomotor, linguistic and social development. In the present paper, the authors report on the outcome measures of visual acuity and strabismus. Poor acuity was defined as a best-corrected acuity of 6/12 or worse on crowded letters or 6/9 or worse on single letters, at age 4 years. Acuity was measured in 79 infants who were significantly hyperopic and/or anisometropic at 11-12 months of age, 23 who showed hyperopia of +3D but less than +3.5D, 196 control subjects, 14 controls with refractive errors, and 126 others who showed an accommodative lag on screening but were not significantly hyperopic on first retinoscopy. There was a poorer acuity outcome in the untreated group of hyperopes compared to controls (p < 0.0001) and to the children who were compliant in spectacle wear (p < 0.001) or who were prescribed spectacles (p < 0.05). Children who were significantly hyperopic at eight months were also more likely to be strabismic by 5.5 years compared to the emmetropic control group (p < 0.001). However, the present study did not find a significant difference in the incidence of strabismus between corrected and uncorrected hyperopic infants. Children who were not refractively corrected for significant hyperopia were four times more likely to have poor acuity at 5.5 years than infants who wore their hyperopic correction, supporting the findings of the First Cambridge Population Infant Vision Screening Programme. PMID- 15545142 TI - Applied force and elongation in the medial rectus in esotropic patients with and without movement restriction. AB - PURPOSE: To study the different ways of expressing the force-elongation relationship in medial rectus muscles in esotropia with and without muscular restriction. SUBJECTS AND METHODS: Twenty-nine passive force-elongation curves were obtained without restriction (group I, n = 13) and with restriction (group II, n = 10) by means of a manual pachymeter and a digital dynamometer. RESULTS: In group I, the mean age was 14 years and 7 days and the mean esotropia was 53.88(Delta) while in group II the mean age was 35 years and 5 days and the mean esotropia was 60.5(Delta). Comparisons of structural muscular parameters between groups I and II were made for length (38.69 +/- 0.75 vs. 32.48 +/- 1.84 mm, p < 0.05), width (8.64 +/- 0.75 vs. 7.95 +/- 0.68 mm), thickness (0.67 +/- 0.07 vs. 0.71 +/- 0.14 mm), and area (5.79 +/- 0.73 vs. 5.62 +/- 1.23 mm2). The differences in width were statistically significant between both groups. As expected, the force-elongation relationship, whether normalized or not, followed an exponential curve. The constant c, which represents force when the elongation is zero, remained the same in all curves. In contrast, the constant b, which represents the slope of the curve, showed a significant difference between the two groups only for the curves of force-absolute elongation and tension-absolute elongation. CONCLUSION: The results imply that the constant b is better for characterizing the difference between the behavior of the medial rectus in esotropia with and without restriction. In addition, the elongation normalization showed that the contractile component is similar between the two groups and, therefore, the classical way of analysis, which does not employ normalization, is appropriate to correlate muscle properties with clinical findings. PMID- 15545143 TI - Does Botulinum toxin have a role in the treatment of small-angle esotropia? AB - A retrospective review of the Toxin Clinic database was carried out over an 18 year period. We identified 68 patients with esotropia of 20 prism dioptres and under who were treated with Botulinum toxin. There were 47 females and 21 males, with a mean age of 34 years (range 11-78 years). Thirty-two (47%) patients had residual esotropia, 20 (29%) had primary esotropia, 11 (16%) had consecutive esotropia, 4 (6%) had secondary esotropia and 1 (1.5%) had a decompensating esophoria. The mean pre-toxin angle was 16 prism dioptres (range 6-20). A total of 434 injections were given, with an average of 6 per patient (range 1-36). Forty-five (66%) patients underwent continued toxin treatment with an interval of 3 to 31 months. Thirteen (19%) patients achieved long-term benefit from only one injection. Seven were unable to demonstrate binocular vision pre-injection but demonstrated it post-injection. Following an initial injection, nine (13%) patients proceeded to surgery. Botulinum toxin was a successful treatment for these patients and was well tolerated, with no side effects. It appears to have a definite role in the treatment of small-angle esotropia. PMID- 15545144 TI - Comparison between two hand-held autorefractors: the Sure-Sight and the Retinomax. AB - PURPOSE: To compare the results of manifest refraction obtained with two different hand-held autorefractors (Sure-Sight, Welch Allyn Co. and Retinomax 1, Nikon Inc.) and with the Topcon RMA 6000 on-table autorefractor in order to estimate any potential bias between these refractometers and to compare the diagnostic performances of these two hand-held autorefractors as screening devices. METHODS: Ninety-eight children were refracted under manifest conditions with the three above-mentioned refractometers and under cycloplegic conditions with the Topcon on-table autorefractor, or by means of retinoscopy. The agreement between the manifest measurements obtained with the three different autorefractors was studied using the method of Bland and Altman. The validity of several thresholds of manifest refractive anomalies as measured with the Sure Sight and with the Retinomax was estimated by receiver operating characteristic (ROC) curves using cycloplegic measures as reference. results There is a spherical positive bias of 1 D between the Sure-Sight and the Retinomax and better agreement between the Topcon and the Retinomax. The surface area indexes of the ROC curves and the diagnostic performances in term of sensitivity and specificity are better with the Retinomax in cases of hyperopia, astigmatism and anisometropia. For myopia, the Sure-Sight has better performance. CONCLUSIONS: The results suggest that either device may be sufficient for assessing refractive errors in children in a screening setting. However, because of a bias between both refractometers in measuring the sphere, distinctive referral criteria must be chosen for the detection of hyperopia and myopia. The diagnostic performance is slightly in favor of the Retinomax. PMID- 15545145 TI - Regarding a new type of eyeglasses for patients with strabismus: The use of prismatic lenses for the cure of squint, tested physiologically. 1847-1848. PMID- 15545147 TI - Cardiovascular complications of COX2 selective inhibitors cause considerable concern. PMID- 15545148 TI - Contemplating the history of drug therapy for hypertension. PMID- 15545149 TI - Essential hypertension in women. PMID- 15545150 TI - The effect of clothes on sphygmomanometric and oscillometric blood pressure measurement. AB - AIM: We determined the effect of wearing clothes under the manometer's cuff on blood pressure in manual auscultatory sphygmomanometric and automatic oscillometric blood pressure measurement. METHODS: Two hundred and one subjects were examined with the auscultatory sphygmomanometric and the automatic oscillometric method, each with and without sleeved arm in random order. The auscultatory readings were blinded for the subjects' state of clothing. Common shirts and sweaters (thinner than 2 mm) were used. RESULTS: Based on confidence intervals of the differences between sleeved and non-sleeved arm measurements and equivalence test, sleeves did not lead to statistically significant effects. Measurements with and without sleeve can be accepted equal within an a priori defined interval of equivalence of +/-4 mmHg. DISCUSSION: This study shows that measuring blood pressure with the manometer's cuff over the subject's sleeve does not differ significantly from non-sleeved arm measurements. This is true for a sample that includes normotensive as well as hypertensive persons with a wide age range. For clinical practice, the not significant mean differences of 0.5-1.1 mmHg are interpreted as not relevant. In this study with a statistical power to find a difference of 4 mmHg, blood pressure measurements were found to be equivalent with and without clothes thinner than 2 mm. PMID- 15545151 TI - The coexistence of acute aortic dissection with autosomal dominant polycystic kidney disease--description of two hypertensive patients. AB - BACKGROUND: Acute aortic dissection is a rare but potentially lethal disease characterized by high early mortality when left untreated. However, survival may be significantly improved by the timely institution of appropriate medical and/or surgical therapy. Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common hereditary disorders, accounting for 8-10% of the cases of end stage renal disease. Hypertension occurs frequently and is an early manifestation of ADPKD in approximately 50-70% of non-azotemic patients. ADPKD, often caused by mutations in the PKD1 gene, is associated with life-threatening vascular abnormalities that are commonly attributed to the frequent occurrence of hypertension. METHODS: We present two cases of hypertensive patients with acute aortic dissection (type A Stanford) coexisting with ADPKD. RESULTS: A 54-year-old male hypertensive patient and a 47-year-old male hypertensive patient, both with known ADPKD were diagnosed and operated with acute dissection of ascending aorta. Both patients were characterized by severe hypertension; therefore, in both cases, degree of hypertension is likely to be causative for this vascular complication. However, since ADPKD is associated with the vascular complications mentioned above, abnormality of the structure of the arterial wall of the thoracic aorta cannot be ruled out. CONCLUSION: Aortic dissection is a rare but potentially disastrous complication, and clinicians should always consider this when dealing with patients with ADPKD who present with chest pain or collapse. PMID- 15545152 TI - Plasma catecholamines, blood pressure responses and perceived stress during mental arithmetic stress in young men. AB - We assessed plasma noradrenaline (NA) and adrenaline (A) at rest during a hyperinsulinaemic glucose clamp and responses to a mental arithmetic stress test (MST) in relation to blood pressure (BP) responses (Finapres) and distress in 20 men with high (> or =140/90 mmHg) and 21 men with normal (< or =115/75 mmHg) screening BP, 21-24 years of age. Perceived stress, effort and overall discomfort were scored 1-10. Catecholamines and BP increased in both groups, change in diastolic BP (DeltaDBP; 9.9 vs. 3.8 mmHg, p < 0.05) and DeltaDBP carryover (recovery period minus baseline) (7.2 vs. 2.2 mmHg, p < 0.01) being greater in men with high screening BP. Independently of BP status, change in systolic BP (DeltaSBP) and DeltaSBP carryover were related to A (both p < 0.001), and DeltaDBP and DeltaDBP carryover to DeltaNA (both p < 0.001). The subjective score sum correlated with maximal NA (rs = 0.40) and A (rs = 0.37) (both p < 0.05). Maximal NA was independently related to stress (p < 0.05) and the subjective score sum (p < 0.01). DeltaA% was greater in the high- (score > or =6) than in the low-stress category, independently of BP status (p < 0.05). High screening BP is associated with impaired BP recovery after mental stress. Plasma catecholamine responses are related to BP responses and carryover effects, and reflect perceived stress in young men. PMID- 15545153 TI - Prognostic value of lipoprotein fractions in essential hypertension. AB - BACKGROUND: To evaluate distribution and prognostic value of total cholesterol and lipoprotein fractions in essential hypertension. METHODS: In a prospective cohort study, 2649 initially untreated subjects with essential hypertension (aged 51, 46.5% women) were investigated at entry and followed for a mean of 5.6 years (range: 1-16). RESULTS: At entry, subjects with total cholesterol (TC) > or =240 mg/dl (> or =6.22 mmol/l) or high-density lipoprotein (HDL) cholesterol (HDL-C) <40 mg/dl (1.05 mmol/l) or low-density lipoprotein (LDL) cholesterol (LDL-C) > or =160 mg/dl (4.13 mmol/l) or TC/HDL-C ratio >6 were 47.7%. TC, HDL-C, LDL-C and triglycerides (TG) did not show any association with office or 24-h ambulatory blood pressure (BP). During follow-up there were 167 first cardiac events and 122 first cerebrovascular events. TC, HDL-C, LDL-C and TC/HDL-C ratio showed an association with cardiac events, but not with cerebrovascular events. TG did not show any association with cardiac or cerebrovascular events. After adjustment for age, sex, diabetes, smoking, left ventricular (LV) hypertrophy and 24-h pulse pressure, the hazard ratio for cardiac events was 1.83 (95% CI 1.23-2.71) in association with a TC > or =6.22 mmol/l, 2.23 with a HDL-C <1.05 mmol/l (95% CI 1.06-4.70), 2.83 with a LDL-C > or =4.91 mmol/l (95% CI 1.48-5.42) and 3.90 with a TC/HDL-C ratio >6.0 (95% CI 2.23-6.81). When forced in the same model, HDL-C and LDL-C showed an independent association with cardiac events. CONCLUSIONS: Abnormalities of TC and lipoproteins are common in essential hypertension. HDL-C and LDL-C independently predict the risk of cardiac, but not cerebrovascular, events. Their predictive value is independent of several confounding factors including LV hypertrophy and ambulatory BP. PMID- 15545154 TI - Cerebral hemodynamics and silent cerebral white matter lesions in middle-aged essential hypertensive patients. AB - Cerebral white matter lesions (WML) represent a subclinical form of ischemic brain damage that have been associated with risk of future stroke. Studies have shown an association between WML and impaired cerebral autoregulation in hypertensives who had previously suffered a stroke. The aim of the study was to evaluate cerebral hemodynamics in asymptomatic hypertensives according to the presence or absence of WML. Fifty never-treated essential hypertensives (32 men, 18 women), aged 50-60 years, without clinical evidence of target organ damage were studied. All patients underwent 24-h ambulatory blood pressure monitoring, and brain-magnetic resonance imaging to establish the presence or absence of WML. Baseline cerebral blood flow velocity (CBF), pulsatility index (PI; differences between systolic and diastolic velocities), and CBF after acetazolamide infusion (vasomotor reactivity of cerebral vessels), were measured by transcranial Doppler ultrasonography in both left and right middle cerebral arteries, and averaged. Twenty hypertensive patients (40%) were found to have WML on brain resonance. No differences were observed on resting and stimulating CBF between hypertensives with and without WML. In contrast, patients with WML exhibited significantly higher PI compared with hypertensives without WML (0.79 +/- 0.13 vs 0.66 +/- 0.12; p = 0.003). Moreover, PI correlated with 24-h pulse pressure (r = 0.361; p = 0.015). We conclude that the presence of silent WML in middle-aged hypertensives is associated with increased cerebrovascular pulsatility. This increased pulsatility is also associated with higher pulse pressure values, suggesting a pathogenetic link between pulse pressure, pulsatility and the development of WML. PMID- 15545155 TI - Effects of nifedipine GITS and diuretics in isolated systolic hypertension--a subanalysis of the INSIGHT study. AB - AIMS: This study tested the effects on cardiovascular outcomes of treatments based on nifedipine gastrointestinal therapeutic system (GITS) compared with the diuretic combination co-amilozide in a pre-specified subset of patients with isolated systolic hypertension (ISH) enrolled in the International Nifedipine GITS Study: Intervention as a Goal in Hypertension Treatment (INSIGHT) study. MAJOR FINDINGS: Of 6321 randomized patients, 1498 (23.7%) had ISH with a baseline mean BP of 173/88 mmHg in both treatment groups. Mean BP fell by 29/10 mmHg in the nifedipine and 30/10 mmHg in the diuretic group to a mean BP of 144/78 mmHg and 143/79 mmHg, respectively, at endpoint. The percentage of primary outcomes in patients with ISH was not significantly different between the two treatment groups (nifedipine GITS 6.0%, co-amilozide 6.6%). The number of ISH patients with composite secondary outcomes was 90 (12.2%) in the nifedipine GITS group and 110 (14.5%) in the co-amilozide group (not significant). The incidence rates of primary and secondary outcomes were similar in patients without ISH. CONCLUSION: In patients with ISH, nifedipine GITS and co-amilozide had similar effects on clinical outcomes and BP lowering. They lend support to international guidelines for the treatment of hypertension recommending the use of long-acting dihydropyridine calcium-channel blockers as one treatment option for patients with ISH. PMID- 15545156 TI - Carotid chemoreceptor reflexes following dietary salt loading in rats. AB - There is a strong association between salt intake and hypertension. Alterations in baroreceptor activity, which precede and contribute to the elevation in blood pressure, have also been shown to affect chemoreceptor reflex response. Dietary salt loading with 8% sodium chloride was carried out in Sprague Dawley rats aged 8 weeks for a period of 5-6 weeks. Blood pressure was thereafter recorded under anaesthesia from the common carotid artery with a Grass Polygraph 7D model, whereas serum Na and K concentrations were measured using a flame photometer. Salt loading resulted in elevated arterial blood pressure as well as hypokalaemia. Stimulation of the carotid chemoreceptor by injection of sodium dithionite resulted in elevated arterial blood pressure, decreased heart rate and hyperventilation in both control and salt-loaded rats. However, the bradycardic response as estimated by the difference in percentage reduction in heart rate was significantly higher in salt rats (36%) than in the control rats (10%). The results indicate that a high-salt diet results in enhanced bradycardic response to carotid chemoreceptor stimulation and that this observation may be related to the attendant hypokalaemia. PMID- 15545158 TI - Exocrine insufficiency in acute pancreatitis. PMID- 15545159 TI - Prevalence of gastro-oesophageal reflux symptoms and the influence of age and sex. AB - BACKGROUND: Most previous studies of reflux symptom prevalence are of small sample size. No reliable data concerning age- and sex-stratified prevalence are available. METHODS: Among 65,363 adult participants in a public health survey in Nord-Trondelag, Norway, 58,596 (90%) responded concerning occurrence and severity of heartburn or regurgitation during the past 12 months. The prevalence of minor, severe and any reflux symptoms was calculated, including stratification for age and sex. In order to examine whether the relative risk of reflux symptoms between sexes, in different age groups, was affected by other potential risk factors for reflux, confounding effects were tested using multivariate logistic regression. Odds ratios and their 95% confidence intervals were used to estimate relative risks. RESULTS: Total prevalence of reflux symptoms was 31.4%, whereof 26.0% were minor symptoms and 5.4% severe symptoms. The prevalence of symptoms occurring at least weekly was 11.6%. Among women, the prevalence increased gradually from 22.1% in the youngest age category to 37.5% in the oldest, while among men it gradually increased from 25.8% in the youngest age group to peak at 36.0% between the ages of 50 and 60 years, after which it declined to 33.8% after age 70. A higher prevalence among women compared to men in the oldest age groups was not explained by confounding by body mass, tobacco smoking, alcohol consumption, dietary factors, or physical exercise. CONCLUSIONS: About every third adult person suffered from reflux symptoms. The prevalence increases linearly with age among women, while among men it peaked between the age of 50 and 70 years and thereafter declined. PMID- 15545160 TI - Significance of cell-surface expression of matrix metalloproteinases and their inhibitors on gastric epithelium and infiltrating mucosal lymphocytes in progression of Helicobacter pylori-associated gastritis. AB - BACKGROUND: Matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) have recently been shown to be important in tissue breakdown and remodeling in gut with inflammatory bowel disease. The role of MMPs and TIMPs remains largely unexplored in Helicobacter pylori-associated gastritis (HAG). The aim of this study was to investigate the expression of these proteolytic enzymes in HAG. METHODS: Cell-surface or/and intracellular expression of MMP-2, 7, 9 and MT1-MMP and TIMPs (TIMP-2 and -4) was determined in gastric epithelium and infiltrative mucosal lymphocytes (IML) in single endoscopic biopsies from H. pylori-infected (n = 25) and uninfected (n = 15) patients. The quantitative analysis was based on the percentage of positive cells detected by flow cytometry. RESULTS: Secreted MMPs and TIMPs as well as membrane type 1-MMP were shown to be localized mainly on the cell surface of both epithelial cells and IML in HAG. H. pylori significantly up-regulated the cell-surface expression not only of MMPs, but also of TIMPs on IML within tissues. The expression of these molecules on IML was correlated with the grade of gastritis. CONCLUSIONS: MMPs and TIMPs expressed on gastric epithelium and H. pylori-antigen(s)-stimulated IML may be implicated in mucosal degradation and remodeling of the stomach. These might contribute to the pathogenesis and progression of atrophy and intestinal metaplasia of gastric mucosa. PMID- 15545161 TI - Correlations of hemoglobin index (IHb) of gastric mucosa with Helicobacter pylori (H. pylori) infection and inflammation of gastric mucosa. AB - BACKGROUND: Helicobacter pylori (H. pylori) infection causes various gastric diseases, among them H. pylori-associated gastritis characterized by diffuse redness of the gastric mucosa. The haemoglobin index (IHb) of the fundic mucosa is an objective parameter of the extent of mucosal redness, but it is unclear whether or not IHb can be used as a diagnostic marker for H. pylori infection. The purpose of this investigation was to evaluate the correlations between IHb of the fundic mucosa and H. pylori infection, inflammatory cell infiltration, and inflammatory mediator production. METHODS: IHb of the fundic mucosa was measured in 108 patients with various gastric diseases (group 1), and values were compared between H. pylori-positive and H. pylori-negative patients. Fifteen patients with H. pylori infection from group 1 underwent H. pylori eradication therapy and IHb was measured before and after treatment. Both IHb and inflammatory cell infiltration were assessed in 61 patients (group 2). In 31 patients from group 2, the expression of interleukin (IL)-8 and inducible nitric oxide synthase (iNOS) messenger RNA (mRNA) was assayed in gastric biopsy specimens by the reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: IHb levels were significantly higher in H. pylori-positive patients than in H. pylori-negative patients (P < 0.001). IHb was decreased at one month after the eradication of H. pylori (P < 0.001). IHb was higher in patients with infiltration by both mononuclear cells and neutrophils (P < 0.001). There was a significant correlation between the IHb level and the expression of IL-8 mRNA (P < 0.001), as well as between IHb and iNOS mRNA expression (P < 0.05). CONCLUSIONS: There were significant correlations between IHb of the gastric mucosa and H. pylori infection, inflammatory cell infiltration, and IL-8/iNOS mRNA expression, suggesting that IHb is a reliable marker of H. pylori infection for use during follow-up endoscopy after H. pylori eradication therapy. PMID- 15545162 TI - Reflux esophagitis after eradication of Helicobacter pylori is associated with the degree of hiatal hernia. AB - BACKGROUND: Several studies have shown that reflux esophagitis (RE) occurs after eradication of Helicobacter pylori. However, endoscopic findings do not allow prediction of the development of RE after successful treatment. In this study, we evaluated the relationship between the prevalence of RE after eradication therapy and the degree of hiatal hernia. METHODS: The study comprised 148 patients who had undergone H. pylori eradication therapy over the past 5 years. The degree of RE and hiatal hernia was evaluated based on endoscopic findings. Hiatal hernia was graded according to Hill's gastroesophageal flap valve (GEFV; grades I-IV) classification. RE after eradication therapy was graded according to the Los Angeles classification system. H. pylori infection was confirmed in all patients by culture, urease test and histological examination of antral and fundic biopsy specimens. RESULTS: Among 148 patients, there were 122 patients (82.4%) with successful and 26 (17.6%) with failed eradication therapy. RE was diagnosed in 25 (20.5%) out of 122 patients with successful therapy but only in 1 (3.8%) out of 26 patients with failed therapy (P < 0.05). After successful eradication, 25 patients had mild RE (12 with grade A, 13 with grade B). Among patients of the successful eradication group (n = 122), RE was diagnosed in 2 (5.3%) out of 38 patients without hiatal hernia and in 23 (27.4%) out of 84 patients with hiatal hernia (P = 0.0051). Furthermore, RE was diagnosed in 2 (5.3%) out of 38 patients with GEFV grade I, 13 (24.1%) out of 54 with grade II, 7 (30.4%) among 23 with grade III, and 3 (42.9%) out of 7 patients with grade IV. The pH level of gastric juice after eradication therapy was lower in the group with successful eradication than in the group with failed therapy regardless of the incidence and degree of RE. CONCLUSIONS: There is a high incidence of RE after successful H. pylori eradication therapy. This incidence of RE was closely associated with the presence and degree of hiatal hernia and with the decrease in gastric juice pH. These findings suggest that the presence of hiatal hernia together with increase in gastric acidity are important determinant factors for the development of RE after successful H. pylori eradication therapy. PMID- 15545163 TI - Long-term effects of eradication of Helicobacter pylori on relapse and histology in gastric ulcer patients: a two-year follow-up study. AB - BACKGROUND: The main purpose of this study was to compare omeprazole (ome) plus two antibiotics (OMC) with omeprazole plus placebo (OP) with regard to gastric ulcer relapse for a period of 2 years in patients who were Helicobacter pylori positive at inclusion. METHODS: Using double-blind randomization 125 patients with gastric ulcer were treated with either OMC (ome 20 mg b.i.d., metronidazole 400 mg b.i.d., clarithromycin 250 mg b.i.d.) (n = 64) or OP (ome 20 mg and placebo) (n = 61) for 1 week, followed by ome 20-40 mg o.d. until healing was confirmed endoscopically after 4, 8 or 12 weeks. Endoscopy and H. pylori diagnostics using culture, histology and serology were performed 6, 12 and 24 months after treatment or at symptomatic relapse. At inclusion, 35% of the OMC group and 38% of the OP group were taking non-steroidal anti-inflammatory drugs (NSAIDs). Nine percent (11/125) of the ulcers were malignant. RESULTS: The prevalence of H. pylori was 82% and the eradication rate 88% in the OMC group and 3% in the OP group. More than 90% of the ulcers were healed after 12 weeks. After 2 years, 76% of patients in the OMC group were in remission compared with 28% in the OP group (ITT) (P < 0.001). Sixty percent of patients in the OMC group that continued to take NSAIDs were in remission after 2 years compared with none in the OP group. Atrophy but not intestinal metaplasia decreased after treatment. CONCLUSIONS: Gastric ulcers are mainly caused by H. pylori, and relapse is effectively prevented by H. pylori eradication, even in patients on NSAIDs. PMID- 15545164 TI - Accuracy of a monoclonal antibody-based stool antigen test in the diagnosis of Helicobacter pylori infection. AB - BACKGROUND: Recent availability of tests for Helicobacter pylori antigens in stool samples has provided potentially useful tools for epidemiological studies and clinical settings. The aim of this study was to evaluate a monoclonal antibody-based H. pylori antigen stool test in the primary diagnosis of H. pylori infection, and to study the test performance after patients were treated with lanzoprazole, and after eradication therapy. METHODS: The study included 122 dyspeptic patients. At gastroscopy, biopsy specimens were obtained for culture and histology. Stool antigen and [14C]-urea breath tests were performed concurrently. Positive culture alone or a positive [14C]-urea breath test in combination with positive histology defined the reference standard. Forty-three Hp +ve patients were treated with lanzoprazole for 2 to 4 weeks, and stool antigen tests were performed on days 1 and 7 post-treatment. After eradication therapy, 32 patients were re-examined for H. pylori infection. RESULTS: Prevalence of H. pylori was 44.3%. Sensitivity and specificity for the stool antigen test in the primary diagnosis of H. pylori infection were 98% and 94%, with positive and negative likelihood ratios of 16.7 and 0.02, respectively. All patients had positive stool tests immediately after lanzoprazole treatment, whereas 2 patients had negative stool tests after 7 days. Triple therapy rendered all patients stool test negative. CONCLUSIONS: The monoclonal antibody-based stool antigen test is an accurate tool in the primary diagnosis of H. pylori infection and after eradication therapy. Lanzoprazole treatment does not influence the clinical performance of the test. PMID- 15545165 TI - IgA antibodies to human tissue transglutaminase: audit of routine practice confirms high diagnostic accuracy. AB - BACKGROUND: The diagnostic accuracy of IgA tissue transglutaminase antibodies (TGA) for coeliac disease (CD) has been assessed following the introduction of the test into routine practice in 2002. METHODS: Specificity was assessed in serum samples received from 1554 adults for routine coeliac serology. The population for assessing sensitivity was 75 consecutive new adult diagnoses of CD. TGA was measured by enzyme-linked immunoassay using human tissue transglutaminase as antigen. Concordance between TGA and endomysial antibody (EMA) was also assessed. RESULTS: The prevalence of new diagnoses of CD in the population tested was 2.8% with similar proportions of new diagnoses in males and females. The positive predictive values at a cut-off of 3 units/mL were 0.77 for samples from Primary Care and 0.92 for samples from Hospital sources, with a sensitivity of 92%. At TGA <3 units/mL, EMA was usually negative; when TGA was >4.9 units/mL, EMA was rarely negative. CONCLUSIONS: We have assessed the role of TGA in routine clinical practice and confirmed high diagnostic accuracy. Sensitivity (92%) is identical to the sensitivity for EMA. IgA deficiency should be excluded in samples showing low absorbance readings in the TGA assay and interference from monoclonal and polyclonal IgA should be excluded in samples with slightly raised TGA levels and negative EMA. TGA is recommended as the first line serological test for coeliac disease. PMID- 15545166 TI - Increased frequency of the immunoglobulin enhancer HS1,2 allele 2 in coeliac disease. AB - BACKGROUND: Coeliac disease (CD) is characterized by increased immunological responsiveness to ingested gliadin in genetically predisposed individuals. This genetic predisposition is not completely defined. A dysregulation of immunoglobulins (Ig) is present in CD: since antiendomysium antibodies (anti-EMA) are of the IgA class. One polymorphic enhancer within the locus control region (LCR) of the immunoglobulin heavy chain cluster at the 3' of the C alpha-1 gene was investigated. The correlation of the penetrance of the four different alleles of the HS1,2-A enhancer of the LCR-1 3' to C alpha-1 in CD patients compared to a control population was analysed. METHODS: A total of 115 consecutive CD outpatients, on a gluten-free diet, and 248 healthy donors, age- and sex-matched, from the same geographical area were enrolled in the study. HS1,2-A allele frequencies were investigated by nested polymerase chain reaction (PCR). RESULTS: The frequency of allele 2 of the enhancer HS1,2-A gene was increased by 30.8% as compared to the control frequency. The frequency of homozygosity for allele 2 was significantly increased in CD patients. Crude odds ratio (OR) showed that those with 2/2 and 2/4 (OR 2.63, P < 0.001 and OR 2.01, P = 0.03) have a significantly higher risk of developing the disease. In contrast, allele 1/2 may represent a protective genetic factor against CD (OR 0.52, P = 0.01). CONCLUSIONS: These data provide further evidence of a genetic predisposition in CD. Because of the Ig dysregulation in CD, the enhancer HS1,2-A may be involved in the pathogenesis. PMID- 15545167 TI - Reduced joint pain after short-term duodenal administration of seal oil in patients with inflammatory bowel disease: comparison with soy oil. AB - BACKGROUND: Rheumatic joint pain is a common extra-intestinal complication of inflammatory bowel disease (IBD). Because the high ratio of n-6 to n-3 fatty acids (FAs) of the Western diet might promote rheumatic disorders, we sought to compare the effects of short-term duodenal administration of n-3-rich seal oil and n-6-rich soy oil on IBD-related joint pain. METHODS: Nineteen patients with IBD-related joint pain were included in the study; 9 had Crohn disease and 10 had ulcerative colitis. Ten millilitres seal oil (n = 10) or soy oil (n = 9) was self administered through a nasoduodenal feeding tube 3 times daily for 10 days. RESULTS: Compared with soy oil treatment, seal oil significantly reduced the duration of morning stiffness (P = 0.024), number of tender joints (P = 0.035), intensity of pain (P = 0.025) and the doctor's scoring of rheumatic disease activity (P = 0.025) at the end of the 10-day treatment period. Analysis of the effects as area under the curve (area between the curve and baseline, zero) for the entire period from start of treatment until 6 months' post-treatment suggested a long-lasting beneficial effect of seal oil administration on joint pain, whereas soy oil tended (not significantly) to aggravate the condition. Consistently, the serum ratios of n-6 to n-3 FAs (P < 0.01) and arachidonic acid to eicosapentaenoic acid (P < 0.01) were reduced after treatment with seal oil. CONCLUSION: The results suggest distinctive, differential prolonged effects on IBD-related joint pain of short-term duodenal administration of n-3-rich seal oil (significant improvement) and n-6-rich soy oil (tendency to exacerbation). PMID- 15545168 TI - Collagenase-1 (MMP-1), matrilysin-1 (MMP-7), and stromelysin-2 (MMP-10) are expressed by migrating enterocytes during intestinal wound healing. AB - BACKGROUND: Matrix metalloproteinases (MMPs) play a crucial role in wound healing of the skin, airways, and cornea, but data on MMPs in normal intestinal wound healing is limited. The aim of this study was to clarify the role of collagenase 1 (MMP-1), matrilysin-1 (MMP-7), and stromelysin-2 (MMP-10) in intestinal wound repair and to determine the effect of cytokines on the expression of these MMPs in intestinal epithelial cell lines. METHODS: Surgical specimens from patients with ischemic colitis (n = 5) were used as an in vivo model of intestinal re epithelialization. Fetal ileal explants were used as an ex vivo model. In situ hybridization for MMPs -1, -3, -7, and -10 was performed and immunohistochemical stainings were used to localize MMP-7 and -9 expressing cells. Stainings for cytokeratin and laminin-5 were performed to identify epithelial cells and migrating enterocytes, respectively. Caco-2, HT-29, and WiDr cell lines were treated for 6-48 h with different cytokines (e.g. EGF, KGF, IL-1 beta, TGF-alpha, TNF-alpha, and TGF-beta1) and Taqman real-time quantitative RT-PCR was used to investigate their effect on the expression of MMPs-1, -7, and -10. RESULTS: MMP 7, MMP-10, and MMP-1 were expressed by migrating enterocytes bordering intestinal ulcers in 5/5, 3/5, and 3/5 samples, respectively. In the fetal gut model, MMP-1 and MMP-10 were expressed by migrating enterocytes, but matrilysin-1 expression was not detected. Matrilysin-1 was up-regulated by TNF-alpha and IL-1 beta, and stromelysin-2 by TNF-alpha and EGF in Caco-2 and WiDr cell cultures. MMP-1 was up regulated in Caco-2 cells by TGF-beta, EGF, and IL-1 beta, but only by EGF in WiDR cells. CONCLUSIONS: It is concluded that collagenase-1, stromelysin-2, and matrilysin-1 are involved in intestinal re-epithelialization in vivo and that they are up-regulated by cytokines relevant in wound repair. PMID- 15545169 TI - Monitoring of long-term thiopurine therapy among adults with inflammatory bowel disease. AB - BACKGROUND: The immunosuppressive effects of thiopurine drugs are mainly mediated through their intracellular metabolism into active 6-thioguanine nucleotide (6 TGN) metabolites, which are incorporated into DNA. Erythrocyte 6-TGN (E-6TGN) levels have been proposed as an instrument for monitoring treatment. The aim of the study was to use erythrocyte E-6TGN, methylated mercaptopurine (MeMP) metabolites, and thiopurine methyltransferase (TPMT) measurements in a clinical setting to determine the clinical outcome in relation to thiopurine metabolism. METHODS: Fifty-five adult patients with inflammatory bowel disease were included in a prospective study and followed for 6 months. Metabolite levels were measured and correlated to outcome and AZA/6-MP dose. RESULTS: The E-6TGN level was significantly related to the TPMT genotype (P = 0.008). Patients in disease remission had higher E-6TGN levels than patients with disease activity both at baseline (P < 0.05) and after 6 months (P = 0.02). Active disease was more frequent among subjects with E-6TGN < or = 125 nmol/mmol Hb at baseline (P = 0.04), but not at 6 months. AZA/6-MP drug dose was positively correlated to E MeMP levels (r = 0.48; P < 0.001) and E-MeMP/E-6TGN ratio (r = 0.41; P = 0.002). Dose changes were positively correlated with the changes in E-MeMP levels (P = 0.01) and E-MeMP/E-6TGN ratio (P = 0.03). CONCLUSIONS: E-6TGN level was the only factor in this study related to disease activity, while there was no relationship between AZA/6-MP dose and E-6TGN levels. This finding illustrates the clinical usefulness of E-6TGN monitoring in the evaluation of treatment intensity. PMID- 15545170 TI - Faecal calprotectin and lactoferrin as markers of acute radiation proctitis: a pilot study of eight stool markers. AB - BACKGROUND: Non-invasive diagnostic tools to evaluate the severity of acute, radiation-induced proctitis are not readily available. The faecal excretion of eight markers of gut inflammation was therefore examined. Five proteins and three lipid derivates were analysed in sequential stool samples taken before and during radiation therapy. METHODS: Stool samples from 15 patients with prostate cancer scheduled for radiation therapy were examined. Pretreatment and in-treatment samples (2nd and 6th weeks) were measured by enzyme-linked immunosorbent assay (ELISA) (calprotectin, lactoferrin, transferrin, leukotriene B4, prostaglandin E2, thromboxane B2 and TNF alpha) or nephelometry (alpha 1-antitrypsin). RESULTS: Calprotectin and lactoferrin concentrations increased significantly during radiation treatment (P = 0.0005 and P = 0.019). Transferrin was detected in only 9 out of 45 samples. There were no changes in tumour necrosis factor alpha (TNF alpha), leukotriene B4, prostaglandin E2 and thromboxane B2 during treatment. alpha 1-antitrypsin could not be detected in any sample. CONCLUSIONS: This study indicates that faecal calprotectin and lactoferrin concentrations could be markers of acute, radiation-induced proctitis. Patient compliance and stability of the markers make this a promising method for clinical research. Eicosanoids could be measured in stool samples, but the concentrations did not increase with increasing radiation dose. PMID- 15545171 TI - Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. AB - BACKGROUND: Gastrointestinal toxicity is one of the most serious side effects in the methotrexate (MTX) treatment. However, the mechanism of the toxicity has not been completely clarified, which may be the reason why symptomatic therapy is carried out. On the other hand, the oxidative stress is known to play an important role in various diseases and drug-induced side effects. In this study the focus was on the oxidative stress in order to clarify the mechanism of MTX induced small intestinal damage, especially neutrophil infiltration. METHODS: MTX (20 mg/kg body wt) was administered to rats intravenously. Mucosal homogenates were prepared from the small intestine and used for assay of biochemical parameters, by which induction of oxidative stress and neutrophil infiltration were evaluated. N-acetylcysteine (NAC; 80 mg/kg body wt), an antioxidant or sodium tungstate (tungsten; 0.7 g/kg body wt), an inhibitor of xanthine dehydrogenase (XD)/xanthine oxidase (XO) known as an important source of reactive oxygen species (ROS) was given to rats with MTX to investigate the contribution of ROS to neutrophil infiltration. RESULTS: The MTX treatment of rats induced the oxidative stress in the small intestine. The ROS production was seen preceding an increase of myeloperoxidase activity, which suggested neutrophil infiltration. Both treatments of NAC and tungsten prevented the MTX-induced ROS production and neutrophil infiltration. CONCLUSIONS: These results suggest that oxidative stress plays an important role in the MTX-induced small intestinal damage, especially neutrophil infiltration. Thus, the modulation of oxidative stress would be useful in reducing intestinal damage in MTX treatment. PMID- 15545172 TI - Plant phenolic 4-coumaric acid protects against intestinal inflammation in rats. AB - BACKGROUND: Many plants contain significant amounts of 4-coumaric acid (4CA), a compound with antioxidant properties in vitro and in vivo. The aim of this study was to assess the effects of 4CA pretreatment on DNA oxidative stress induced by intestinal inflammation in rodents. METHODS: 4CA (50 mg/kg) was administered to rats for 14 days mixed in the diet. Colitis was induced on days 13 and 14 by administering 6% (w/v) dextran sodium sulphate (DSS) in the drinking water. RESULTS: In the colon mucosa, DSS treatment increased myeloperoxidase activity (P < 0.05), oxidative DNA damage (P < 0.01), and cyclooxygenase-2 (COX-2) expression (P < 0.01) and reduced superoxide dismutase-2 (SOD-2) expression (P < 0.05). It was found that treatment with 4CA prior to DSS-induced inflammation reduced oxidative DNA damage (P < 0.01), COX-2 over-expression (P < 0.01) and restored SOD-2 gene expression to control levels. Similar effects were observed with nimesulide administered p.o. (5 mg/kg, 1 day before and during DSS treatment). PGE levels in plasma and colon mucosa were increased by DSS treatment and this effect was inhibited by pretreatment with 4-CA (P < 0.01). CONCLUSIONS: Mild acute intestinal inflammation induced by DSS can be inhibited by 4-CA and this action is associated with the suppression of COX-2 expression and activity. PMID- 15545173 TI - Genetic characterization of colorectal cancers in young patients based on chromosomal loss and microsatellite instability. AB - BACKGROUND: In recent studies a high frequency of microsatellite instability among colorectal cancers in young patients has been reported, but the frequency of microsatellite instability (MSI) and chromosomal instability among colorectal cancers in young patients has not yet been fully elucidated. Only one report showed an increased loss of heterozygosity (LOH) ratio at 9p locus, which harbors tumor suppressor genes p16. The LOH and MSI status among colorectal cancers in young patients was examined. METHODS: Twenty-five patients under 40 years of age diagnosed with colorectal cancer were examined for MSI and LOH using 17 microsatellite markers, and also p16 expression patterns were evaluated by immunohistochemistry and methylation status of the p16 gene was assessed by methylation-specific PCR. RESULTS: MSI was observed in only one case (4%). LOH at 2p, 5q, 9p, 11q, 17p, and 18q was observed in 41%, 59%, 42%, 35%, 46%, and 56% of cases, respectively. Eighty-three percent of patients showed p16-positive expression patterns. Fifty percent of colorectal cancers in young patients exhibited p16 methylation (3/6). CONCLUSIONS: Our study demonstrated that colorectal cancers in young patients without MSI showed a high frequency of LOH at the 9p locus. However, LOH status at 9p and p16 expression pattern did not show a significant correlation. Other tumor suppressor genes on the 9p, with the exception of p16, may play an important role in the carcinogenesis of colorectal cancers in young patients. PMID- 15545174 TI - Portal and systemic serum growth factor and acute-phase response after laparotomy or partial hepatectomy in patients with colorectal liver metastases: a prognostic role for C-reactive protein and hepatocyte growth factor. AB - BACKGROUND: Growth factors play a role in wound healing and tumour growth. The aim of this study was to compare the effect of partial hepatectomy (PH) and laparotomy on serum levels of growth factors and acute-phase proteins in patients with colorectal liver metastases and to correlate these levels with prognosis after PH. METHODS: Epidermal growth factor (EGF), hepatocyte growth factor (HGF), insulin like growth factor-I (IGF-I), insulin, interleukin-6 (IL-6), C-reactive protein (CRP) and serum amyloid-A (SAA) were determined in portal and systemic serum in 24 PH patients and 9 laparotomy patients. RESULTS: No differences were found in the clinicopathological characteristics of PH and laparotomy patients with the exception of the number of metastases, blood loss and operation time. The response of SAA, CRP and IGF-I was lower in PH patients than in laparotomy patients (P < 0.02). PH was associated with a higher IL-6 (P = 0.02) and HGF (P = 0.055) response than laparotomy. A higher HGF and CRP response was associated with a poorer prognosis. Total IGF-I was negatively correlated with the resected liver volume (r = -0.48, P < 0.05). CONCLUSIONS: PH is associated with a lower acute-phase and total IGF-I response and a higher HGF and IL-6 response compared with laparotomy. HGF and CRP responses had an influence on the prognosis. PMID- 15545175 TI - Clinical value of increased serum creatinine concentration as predictor of short term outcome in decompensated cirrhosis. AB - BACKGROUND: The purpose of this study was to assess whether serum creatinine concentration alone or associated with other biological parameters was an independent predictor of short-term mortality in patients with decompensated cirrhosis. METHODS: A total of 212 consecutive episodes of decompensated cirrhosis in patients admitted to the hospital between January 1999 and December 2001 were reviewed retrospectively. Depending on a serum creatinine concentration equal to or greater than 1.5 mg/dL at the time of admission, patients were divided into decompensated cirrhosis with renal failure (101 episodes in 59 patients, aged 69.8 +/- 10 years) and without renal failure (111 episodes in 61 patients, aged 64.5 +/- 13 years). Outcome (alive, death) during the episode of decompensation of liver disease and outcome at 90 days after admission were assessed. RESULTS: Differences in the frequency of variables according to outcome in the overall episodes of decompensated cirrhosis with and without renal failure showed significant differences between patients who died and those who were alive both at hospital discharge and at 90 days in serum bilirubin, Child-Pugh score, MELD (model for end-stage liver disease) score, and serum creatinine levels. In the multivariate analysis, serum creatinine was not an independent predictor of outcome. The prediction accuracy according to the area under the ROC (receiver operating characteristic) curve was greater for the MELD scale than for serum creatinine. CONCLUSIONS: Serum creatinine concentration is a parameter that should be included in the prognostic assessment of patients with decompensated cirrhosis, but should be combined with other specific parameters of liver function, such as bilirubin, albumin, and the international normalized ratio (INR) for prothrombin time. PMID- 15545176 TI - Morphological changes after steroid therapy in autoimmune pancreatitis. AB - BACKGROUND: Although many patients with autoimmune pancreatitis undergo steroid therapy, detailed evaluation of morphological changes in the pancreas and bile duct following therapy has not been performed in this disease. In this study serological and morphological changes occurring during steroid treatment of autoimmune pancreatitis are comparatively examined. METHODS: Ten patients with autoimmune pancreatitis were treated with corticosteroids. Morphological findings were: pancreatic enlargement (n = 9), irregular narrowing of the main pancreatic duct (n = 10), and biliary stenosis (n = 9). An initial dose of prednisolone was 40-30 mg/day, and this was tapered by 5 mg every 1-2 weeks. All patients underwent ultrasound and serological testing 1-2 weeks after commencing medication, followed by weekly serological testing and by CT and endoscopic retrograde cholangiopancreatography after 1-2 months. Radiological and serological changes were compared. RESULTS: All 10 patients were responsive to steroid therapy. Pancreatic size normalized within 1 month; however, irregularity of the pancreatic duct remained in 6 patients. Rigidity or lateral deformity of the bile duct remained in 3 patients and biliary stenosis persisted in 5. Four patients in whom elevated serum IgG4 failed to normalize also showed incomplete morphological improvement. Three patients with complete improvement of the pancreatic duct stopped medication, but recurrence of pancreatitis did not occur. CONCLUSIONS: Although steroid therapy was morphologically and serologically effective in patients with autoimmune pancreatitis, cholangiopancreatographic abnormalities remained in many patients. Morphological improvement on cholangiopancreatography and normalization of serum IgG4 after steroid therapy appeared to be good indicators for discontinuing medication in patients with autoimmune pancreatitis. PMID- 15545177 TI - Immunohistochemical analysis of molecular biological factors in intraductal papillary-mucinous tumors and mucinous cystic tumors of the pancreas. AB - BACKGROUND: To investigate the malignancy and differentiation of intraductal papillary-mucinous tumors (IPMTs) and mucinous cystic tumors (MCTs) of the pancreas, clinicopathologic characteristics and immunohistochemical features were analyzed. METHODS: The clinicopathologic characteristics and immunohistochemical features of 24 patients with IPMT and 8 with MCT who underwent pancreatic resections at our hospital were examined. Immunohistochemical features analyzed included expression of p53 protein, proliferating cell nuclear antigen, integrins, interleukin-1 receptor type I, and hormone-associated receptors, and the factors correlated with malignancy were identified by multiple logistic regression. RESULTS: Among the IPMTs, there were 16 intraductal papillary adenomas, 5 intraductal papillary adenocarcinomas, and 3 moderate dysplasias. Among the MCTs, there were 6 mucinous cyst adenomas and 2 mucinous cyst adenocarcinomas. Multivariate analysis revealed that of the clinicopathologic characteristics, only the presence of mural nodules (odds ratio (OR) 7.12, P = 0.044) was independently correlated with the malignancy of IPMTs, and that of the immunohistochemical features, only alpha integrin subunit expression was independently correlated with malignancy of pancreatic mucinous tumors (OR 15.6, P = 0.036), especially IPMTs (OR 35.7, P = 0.012). CONCLUSION: These results indicate that alpha-containing integrin expression can be a significant marker of malignancy in pancreatic mucinous tumors. PMID- 15545178 TI - Decreased leptin production in mice after onset of ulcerative colitis-like disease. PMID- 15545179 TI - Toxic acute hepatitis and hepatic fibrosis after consumption of chaparral tablets. AB - In this report we describe a young, previously healthy woman who developed severe acute hepatitis after consumption of chaparral tablets, a commonly used herbal product. In this case, the elimination-rechallenge event and the exclusion of other possible aetiologic factors strongly supported true causality between the herbal product and the liver damage. Primary liver biopsy showed severe toxic hepatitis consistent with previous reports of chaparral-induced liver damage. Later, 6 months after the liver function tests had normalized, permanent hepatic fibrosis could still be seen. PMID- 15545180 TI - Chronic perineal fistula as a late failure manifestation of pseudomyxoma peritonei syndrome. AB - Pseudomyxoma peritonei syndrome (PMP) is a rare but fascinating entity in surgical oncology. It consists of the accumulation of mucus in the peritoneal cavity, sometimes in huge amounts, in most of the cases after rupture of an appendiceal tumor. The diagnosis and management of PMP has changed significantly in the past 15 years, with the institution of an aggressive therapeutic combination: cytoreductive surgery and intraperitoneal perioperative chemotherapy. Few reports deal with the late manifestations and complications at endstage of the disease. The cases presented here are unique in that they illustrate a late 'failure' in the perianal area in three patients with long-term PMP. PMID- 15545182 TI - Guidelines for the management of gastroenteropancreatic neuroendocrine tumours (including bronchopulmonary and thymic neoplasms). Part I-general overview. AB - The incidence of neuroendocrine tumours of the gastroenteropancreatic system seems to have increased during the past decade. New diagnostic and therapeutic procedures have aroused the interest of physicians, though most see very few cases of such diseases. A group of members of the Nordic Neuroendocrine Tumour Group decided to compile some guidelines to facilitate the diagnosis and treatment of patients with these tumours. Part I of these guidelines discusses the principles of histopathology, biochemical and radiological diagnosis as well as therapeutic options. PMID- 15545183 TI - Guidelines for the management of gastroenteropancreatic neuroendocrine tumours (including bronchopulmonary and thymic neoplasms). Part II-specific NE tumour types. AB - Part II of the guidelines contains a description of epidemiology, histopathology, clinical presentation, diagnostic procedure, treatment, and survival for each type of neuroendocrine tumour. We are not only including gastroenteropancreatic tumours but also bronchopulmonary and thymic neuroendocrine tumours. These guidelines essentially cover basic knowledge in the diagnosis and management of the different forms of neuroendocrine tumour. We have, however, tried to give more updated information about the epidemiology and histopathology, which is essential for the clinical management of these tumours. PMID- 15545184 TI - The need for oncogenetic counselling. Ten years' experience of a regional oncogenetic clinic. AB - A monogenic inheritance, mainly seen as a dominant pattern, accounts for 5-10% of all cancer cases. The increased knowledge and identification of high-risk genes have led to a need for specialized cancer family clinic was the expression used by Eeles and Murday. The Oncogenetic Clinic at the University Hospital in Lund was started in 1993 and the authors' 10-year experience is summarized in this paper. The clinic offers service to the South Swedish Health Care Region comprising a total of 1.6 million inhabitants. During these first 10 years a total of 1059 individuals from 789 families have been individually counselled. The most common reason for referral was a family history of breast cancer, followed by a family history of colorectal cancer. According to the commonly used criteria, 437 (55%) of the families were considered as autosomal dominantly inherited; 147 families (19%) did not fulfil these criteria but had a strong clustering of breast/ovarian or colorectal/endometrial cancer. The remaining 205 families (26%) were not recognized as any previously described hereditary cancer syndrome with early onset. However, most of these families had a family history of cancer. Mutation analysis was performed in 386/789 (49%) of the families. In families with breast and ovarian cancer a genetic aberration was identified in 45/76 (59%) and in breast-only families in 27/129 (21%). In MSI-positive colon cancer families 16/34 (47%) of the families had a germline mutation. Thus, the majority of the families referred to the clinic were in obvious need of genetic counselling concerning cancer and heredity and in a substantial number of the families a germline mutation could be identified. PMID- 15545185 TI - Ten-year follow-up of a randomized controlled trial of adjuvant clodronate treatment in node-positive breast cancer patients. AB - Ten-year follow-up results are presented of an adjuvant clodronate trial in patients with primary breast cancer. Between 1990 and 1993, 299 women with primary node positive breast cancer were randomized to oral clodronate 1600 mg daily (149) or controls (150) for 3 years. All patients received adjuvant chemo- or endocrine therapy. Within 10 years bone metastases were detected at the same frequency in the clodronate and control groups: 44 (32%) vs. 42 (29%), respectively, (p=0.35). The frequency of non-skeletal recurrences (visceral and local) was significantly higher in the clodronate group 69 (50%) as compared with the controls 51 (36%) (p=0.005). Ten-year disease-free survival (DFS) remained significantly lower in the clodronate group (45% vs. 58%, p=0.01, respectively). This was especially seen in oestrogen receptor negative patients (25% vs. 58%, p=0.004, respectively). No significant overall survival difference was found between the groups. As previously reported 3-year adjuvant clodronate treatment did not prevent the development of bone metastases in node-positive breast cancer patients. A negative effect of clodronate on DFS by increasing the development of visceral metastases was still seen at 10 years, but this did not significantly compromise overall survival. PMID- 15545186 TI - Influence of overall treatment time and radiobiological parameters on biologically effective doses in cervical cancer patients treated with radiation therapy alone. AB - The aim of the study was to examine the influence of overall treatment time (OTT) on the value of calculated biological effective doses (BEDs) for different biological variables. These variables were: tumour proliferation rate, different cell radiosensitivity (alpha=0.2, 0.3, and 0.4 /Gy), and different start time for repopulation (Tk=21, 28, and 35 days). Also the influence of age (), Hb level (), tumor proliferation rate (bromodeoxyuridine labelling index; BrdUrdLI), and DNA ploidy on survival after shorter (60 days) OTT was investigated. The study included 229 patients with cervix carcinoma treated entirely by standard radiotherapy (RT) (external beam RT plus low-medium dose-rate (LDR/MDR) brachytherapy (BT) at the Center of Oncology in Krakow. The linear quadratic equation was used to calculate BED, which is proportional to log cell kill. BEDs 10 (for tumours) were calculated with consideration of OTT for each patient and tumour proliferation rate (standardized potential doubling time; standardized Tpot) based on BrdUrdLI assessed on biopsy material before RT. Median OTT was 90 days (range 30-210). The mean calculated total BED for point A for tumour and 'early reactions' was equal to 103.0 Gy10. The longest median survival time--52 months--was seen for patients treated with OTT 8.8%) BED loss was 1.4 Gy/day and for slowly proliferating tumours (BrdUrdLI 50 years (p=0.003) and high Hb level (>116 g/l) (p=0.041). For longer treatments (OTT >60 days) the unfavourable parameters were: age 0.05). There was a consistent trend in all participants for the ergonomic sock to induce a higher core temperature and higher skin temperatures compared to the standard sock. Overall mean ratings of perceived exertion and ratings of thermal perception were similar for both sock conditions. Participant questionnaires highlighted a general perception that the ergonomic socks had superior cushioning but that the standard socks were comfortable to wear. Despite there being no significant physiological or thermal differences between socks, the ergonomic sock was perceived to be cooler and was the preferred sock which suggests that subjective perceptions may be more important than objective measurements when selecting a sock for wear during prolonged exercise. PMID- 15545240 TI - From the editor--drug companies under investigation. PMID- 15545241 TI - An intergenerational group benefits both emotionally disturbed youth and older adults. AB - This qualitative descriptive study examined the perceived benefits of an intergenerational program for low-income older adults residing in subsidized housing and youth who were part of a support group for emotionally disturbed youth. The intergenerational group met bimonthly for 11 months at the independent living facility for the elderly. Activities consisted of group discussions, games, talent expressions, trips, picnics, and crafts. Frequently a teen was paired with an older adult to complete a craft project. Bonding between each pair occurred over time, as the older adults became role models for the youth. The perceived benefits of including emotionally disturbed youth and vulnerable but well older adults in an intergenerational program were determined by a structured interview and then through categorization of the participants' responses to a series of emotional, attitudinal, and behavioral questions using Yalom's (1985)Therapeutic Factors. The results indicated that both of the age groups' attitudes toward each other and relationships with each other changed positively. Behavioral changes among the youth included improved social skills observed by the co-facilitators of the group. PMID- 15545242 TI - Relationships among risk, and communication and social skills in a high security forensic setting. AB - The Behavioural Status Index was developed for risk assessment within forensic care. This paper reports data analysis for the Behavioural Status Index and its subscales. Data were collected, using a repeated measures method by primary nurses, from a sample of 503 individual patients in two high security mental health hospitals in the UK. Results are reported for inter-item correlations, factor analysis, and differences among independent groups of patients, categorized by Mental Health Act 1983 classification, patient ward dependency, and patient gender. Data trends are indicative of clinically interesting relationships. A distinct factorial structure emerged suggesting groupings of behaviours. PMID- 15545243 TI - Embodiment of severe and enduring mental illness: finding meaning in schizophrenia. AB - For many individuals, schizophrenia is a severe and enduring illness. While nurses need to understand the symptomatology of the illness in order to provide specific care and treatment, it also is important to find out how people with schizophrenia embody the illness. Capturing this knowledge will help nurses to provide more appropriate care to these individuals. This paper, which is taken from a larger qualitative study, reports the lived experience of young adults with schizophrenia. Three main themes emerged from the data, which highlighted how these individuals found meaning in schizophrenia. The first theme,"embodied temporality: illness as a catastrophic experience,"portrayed how schizophrenia affected participants' temporality or lived time. The second theme,"embodied relationality: illness as a mediator of social relationships,"reflected how the illness affected their relationships with others. The third theme,"embodied treatment: medications side effects as burdensome,"illustrated how the side effects of antipsychotic medications distorted the individual's perception of his or her body and how it compromised the ability to establish and maintain sexual relationships. The findings are important to mental health nurses because they highlight the need to be sensitive to how people with schizophrenia find meaning in their illness experience and to incorporate this knowledge into the care that they provide. PMID- 15545244 TI - Grief in the shadows: exploring loss and bereavement in people with developmental disabilities. AB - A greater understanding of how developmentally disabled people cope with loss and bereavement is needed to improve assessment and intervention for these individuals. Misconceptions exist about how effectively developmentally disabled persons can articulate their perceptions and feelings about traumatic experiences, death, and the subsequent impact of the events that follow. The lived experience and expression of grief may differ for the developmentally disabled population. As a result, their reactions may not be adequately recognized, interpreted, or managed therapeutically, compounding their distress. Three case reports of developmentally disabled individuals who experienced a loss are presented, along with helpful guidelines regarding assessment and intervention. PMID- 15545245 TI - Belonging and adapting: mental health of Bosnian refugees living in the United States. AB - The purpose of this study was to elucidate the experience of Bosnian refugees currently living in the United States. Using a phenomenological method, seven adult female Bosnian refugees each participated in an audio-recorded interview lasting from one to two hours. Two major themes emerged from the analyses of the text: belonging and adapting. Belonging included concepts of cultural memory, identity and difference, empathy and reciprocity, and perfection of speech. Adapting focused on coping with transitions, coping with memories of past and attendant losses, coping with accepting a new culture while trying to fit into the new culture, and learning the new language perfectly. Implicit in the refugees' experiences were states of culture shock, loneliness, psychic numbness, grief, nostalgia, and feelings of dejection, humiliation, inferiority, and feeling as if they belonged nowhere. Simultaneously, the refugees reported feelings of relief and safety after leaving behind the threat of death in their old homes, feelings of gratefulness for their new freedom to hope for a better life, and their restored ability to notice beauty, as well as a sense of normalcy in their new lives. Recommendations for nursing research include the need to identify additional factors promoting successful belonging and adapting in refugees. Recommendations for nursing practice include the importance of adopting a perspective that is respectful of the uniqueness of each refugee and the necessity for recognizing the normal processes of refugee adaptation. PMID- 15545246 TI - Interrelations of psychiatric patient experiences of boredom and mental health. AB - A review of literature related to psychiatric patients' interrelated experiences of boredom and mental health demonstrated that developing a better understanding of the relationship between boredom and health would provide important implications for development of the therapeutic environment of psychiatric units. The literature suggested that experiences of meaningful roles and relationships contribute substantially to experiences of control essential for mental health. Boredom is an indicator of a lack of experience of meaning. Many psychiatric patients experience boredom and lack opportunities to experience meaning, indicating a lack in the therapeutic potential of the hospital environment and a requisite for change and further research. PMID- 15545248 TI - Bayesian estimation of ancestral character states on phylogenies. AB - Biologists frequently attempt to infer the character states at ancestral nodes of a phylogeny from the distribution of traits observed in contemporary organisms. Because phylogenies are normally inferences from data, it is desirable to account for the uncertainty in estimates of the tree and its branch lengths when making inferences about ancestral states or other comparative parameters. Here we present a general Bayesian approach for testing comparative hypotheses across statistically justified samples of phylogenies, focusing on the specific issue of reconstructing ancestral states. The method uses Markov chain Monte Carlo techniques for sampling phylogenetic trees and for investigating the parameters of a statistical model of trait evolution. We describe how to combine information about the uncertainty of the phylogeny with uncertainty in the estimate of the ancestral state. Our approach does not constrain the sample of trees only to those that contain the ancestral node or nodes of interest, and we show how to reconstruct ancestral states of uncertain nodes using a most-recent-common ancestor approach. We illustrate the methods with data on ribonuclease evolution in the Artiodactyla. Software implementing the methods (BayesMultiState) is available from the authors. PMID- 15545249 TI - Column sorting: rapid calculation of the phylogenetic likelihood function. AB - Likelihood applications have become a central approach for molecular evolutionary analyses since the first computationally tractable treatment two decades ago. Although Felsenstein's original pruning algorithm makes likelihood calculations feasible, it is usually possible to take advantage of repetitive structure present in the data to arrive at even greater computational reductions. In particular, alignment columns with certain similarities have components of the likelihood calculation that are identical and need not be recomputed if columns are evaluated in an optimal order. We develop an algorithm for exploiting this speed improvement via an application of graph theory. The reductions provided by the method depend on both the tree and the data, but typical savings range between 15%and 50%. Real-data examples with time reductions of 80%have been identified. The overhead costs associated with implementing the algorithm are minimal, and they are recovered in all but the smallest data sets. The modifications will provide faster likelihood algorithms, which will allow likelihood methods to be applied to larger sets of taxa and to include more thorough searches of the tree topology space. PMID- 15545250 TI - Multiple data sets, high homoplasy, and the phylogeny of softshell turtles (Testudines: Trionychidae). AB - We present a phylogenetic hypothesis and novel, rank-free classification for all extant species of softshell turtles (Testudines:Trionychidae). Our data set included DNA sequence data from two mitochondrial protein-coding genes and a approximately 1-kb nuclear intron for 23 of 26 recognized species, and 59 previously published morphological characters for a complimentary set of 24 species. The combined data set provided complete taxonomic coverage for this globally distributed clade of turtles, with incomplete data for a few taxa. Although our taxonomic sampling is complete, most of the modern taxa are representatives of old and very divergent lineages. Thus, due to biological realities, our sampling consists of one or a few representatives of several ancient lineages across a relatively deep phylogenetic tree. Our analyses of the combined data set converge on a set of well-supported relationships, which is in accord with many aspects of traditional softshell systematics including the monophyly of the Cyclanorbinae and Trionychinae. However, our results conflict with other aspects of current taxonomy and indicate that most of the currently recognized tribes are not monophyletic. We use this strong estimate of the phylogeny of softshell turtles for two purposes: (1) as the basis for a novel rank-free classification, and (2) to retrospectively examine strategies for analyzing highly homoplasious mtDNA data in deep phylogenetic problems where increased taxon sampling is not an option. Weeded and weighted parsimony, and model-based techniques, generally improved the phylogenetic performance of highly homoplasious mtDNA sequences, but no single strategy completely mitigated the problems of associated with these highly homoplasious data. Many deep nodes in the softshell turtle phylogeny were confidently recovered only after the addition of largely nonhomoplasious data from the nuclear intron. PMID- 15545251 TI - The origin and radiation of Macaronesian beetles breeding in Euphorbia: the relative importance of multiple data partitions and population sampling. AB - Species-level phylogenies derived from many independent character sources and wide geographical sampling provide a powerful tool in assessing the importance of various factors associated with cladogenesis. In this study, we explore the relative importance of insular isolation and host plant switching in the diversification of a group of bark beetles (Curculionidae: Scolytinae) feeding and breeding in woody Euphor biaspurges. All species in the genus Aphanarthrumare each associated with only one species group of Euphorbia(succulents or one of three different arborescent groups), and the majority of species are endemic to one or several of the Macaronesian Islands. Hence, putative mechanisms of speciation could be assessed by identifying pairs of sister species in a phylogenetic analysis. We used DNA sequences from two nuclear and two mitochondrial genes, and morphological characters, to reconstruct the genealogical relationships among 92 individuals of 25 species and subspecies of Aphanarthrumand related genera. A stable tree topology was highly dependent on multiple character sources, but much less so on wide population sampling. However, multiple samples per species demonstrated one case of species paraphyly, as well as deep coalescence among three putative subspecies pairs. The phylogenetic analyses consistently placed the arborescent breeding and West African--Lanzarote-distributed species A. armatumin the most basal position in Aphanarthrum, rendering this genus paraphyletic with respect to Coleobothrus. Two major radiations followed, one predominantly African lineage of succulent feeding species, and one island radiation associated with arborescent host plants. Sister comparisons showed that most recent divergences occurred in allopatry on closely related hosts, with subsequent expansions obscuring more ancient events. Only 6 out of 24 cladogenetic events were associated with host switching, rendering geographical factors more important in recent diversification. PMID- 15545253 TI - Explosive radiations and the reliability of molecular clocks: island endemic radiations as a test case. AB - The reliability of molecular clocks has been questioned for several key evolutionary radiations on the basis that the clock might run fast in explosive radiations. Molecular date estimates for the radiations of metazoan phyla (the Cambrian explosion) and modern orders of mammals and birds are in many cases twice as old as the palaeontological evidence would suggest. Could some aspect of explosive radiations speed the molecular clock, making molecular date estimates too old? Here we use 19 independent instances of recent explosive radiations of island endemic taxa as a model system for testing the proposed influence of rapid adaptive radiation on the rate of molecular evolution. These radiations are often characterized by many of the potential mechanisms for fast rates in explosive radiations--such as small population size, elevated speciation rate, rapid rate of morphological change, release from previous ecological constraints, and adaptation to new niches--and represent a wide variety of species, islands, and genes. However, we find no evidence of a consistent increase in rates in island taxa compared to their mainland relatives, and therefore find no support for the hypothesis that the molecular clock runs fast in explosive radiations. PMID- 15545252 TI - Molecular phylogenetics of squamata: the position of snakes, amphisbaenians, and dibamids, and the root of the squamate tree. AB - Squamate reptiles (snakes, lizards, and amphisbaenians) serve as model systems for evolutionary studies of a variety of morphological and behavioral traits, and phylogeny is crucial to many generalizations derived from such studies. Specifically, the traditional dichotomy between Iguania (anoles, iguanas, chameleons, etc.) and Scleroglossa (skinks, geckos, snakes, etc.) has been correlated with major evolutionary shifts within Squamata. We present a molecular phylogenetic study of 69 squamate species using approximately 4600 (2876 parsimony-informative) base pairs (bp) of DNA sequence data from the nuclear genes RAG-1(approximately 2750 bp) and c-mos(approximately 360 bp) and the mitochondrial ND2 region (approximately 1500 bp), sampling all major clades and most major subclades. Under our hypothesis, species previously placed in Iguania, Anguimorpha, and almost all recognized squamate families form strongly supported monophyletic groups. However, species previously placed in Scleroglossa, Varanoidea, and several other higher taxa do not form monophyletic groups. Iguania, the traditional sister group of Scleroglossa, is actually highly nested within Scleroglossa. This unconventional rooting does not seem to be due to long branch attraction, base composition biases among taxa, or convergence caused by similar selective forces acting on nonsister taxa. Studies of functional tongue morphology and feeding mode have contrasted the similar states found in Sphenodon(the nearest outgroup to squamates) and Iguania with those of Scleroglossa, but our findings suggest that similar states in Sphenodonand Iguania result from homoplasy. Snakes, amphisbaenians, and dibamid lizards, limbless forms whose phylogenetic positions historically have been impossible to place with confidence, are not grouped together and appear to have evolved this condition independently. Amphisbaenians are the sister group of lacertids, and dibamid lizards diverged early in squamate evolutionary history. Snakes are grouped with iguanians, lacertiforms, and anguimorphs, but are not nested within anguimorphs. PMID- 15545254 TI - Molecular systematics and biogeography of the southern South american freshwater "crabs" Aegla (decapoda: Anomura: Aeglidae) using multiple heuristic tree search approaches. AB - Recently new heuristic genetic algorithms such as Treefinder and MetaGA have been developed to search for optimal trees in a maximum likelihood (ML) framework. In this study we combined these methods with other standard heuristic approaches such as ML and maximum parsimony hill-climbing searches and Bayesian inference coupled with Markov chain Monte Carlo techniques under homogeneous and mixed models of evolution to conduct an extensive phylogenetic analysis of the most abundant and widely distributed southern South American freshwater"crab,"the Aegla(Anomura: Aeglidae). A total of 167 samples representing 64 Aegla species and subspecies were sequenced for one nuclear (28S rDNA) and four mitochondrial (12S and 16S rDNA, COI, and COII) genes (5352 bp total). Additionally, six other anomuran species from the genera Munida,Pachycheles, and Uroptychus(Galatheoidea), Lithodes(Paguroidea), and Lomis(Lomisoidea) and the nuclear 18S rDNA gene (1964 bp) were included in preliminary analyses for rooting the Aegla tree. Nonsignificantly different phylogenetic hypotheses resulted from all the different heuristic methods used here, although the best scored topologies found under the ML hill-climbing, Bayesian, and MetaGA approaches showed considerably better likelihood scores (Delta> 54) than those found under the MP and Treefinder approaches. Our trees provided strong support for most of the recognized Aegla species except for A. cholchol,A. jarai,A. parana,A. marginata, A. platensis, and A. franciscana, which may actually represent multiple species. Geographically, the Aegla group was divided into a basal western clade (21 species and subspecies) composed of two subclades with overlapping distributions, and a more recent central-eastern clade (43 species) composed of three subclades with fairly well-recognized distributions. This result supports the Pacific-Origin Hypothesis postulated for the group; alternative hypotheses of Atlantic or multiple origins were significantly rejected by our analyses. Finally, we combined our phylogenetic results with previous hypotheses of South American paleodrainages since the Jurassic to propose a biogeographical framework of the Aegla radiation. PMID- 15545255 TI - Testing nested phylogenetic and phylogeographic hypotheses in the Plethodon vandykei species group. AB - Mesic forests in the North American Pacific Northwest occur in two disjunct areas: along the coastal and Cascade ranges of Oregon, Washington, and British Columbia as well as the Northern Rocky Mountains of Idaho, Montana, and British Columbia. Over 150 species or species complexes have disjunct populations in each area, and a priori hypotheses based on phytogeography and geology potentially explain the disjunction via either dispersal or vicariance. Here, we test these hypotheses in the disjunct salamander complex Plethodon vandykei and P. idahoensisby collecting genetic data (669 bp of Cyt b) from 262 individuals. Maximum likelihood analysis indicated reciprocal monophyly of these species, supporting the ancient vicariance hypothesis, whereas parametric bootstrap and Bayesian hypothesis testing allow rejection of the dispersal hypothesis. The coalescent estimate of the time since population divergence (estimated using MDIV) is 3.75 x 106 years, and the 95%credibility interval of this value overlaps with the geological estimate of vicariance, but not the hypothesized dispersal. These results are congruent with the pattern seen in other mesic forest amphibian lineages and suggest disjunction in amphibians may be a concerted response to a geological/climatological event. WithinP. idahoensis, we tested the corollary hypothesis of an inland Pleistocene refugium in the Clearwater drainage with nested clade analysis and coalescent estimates of population growth rate (g). Both analyses support post-Pleistocene expansion from the Clearwater refugium. We corroborated this result by calculating Tajima's Dand mismatch distribution within each drainage, showing strong evidence for recent population expansion within most drainages. This work demonstrates the utility of statistical phylogeography and contributes two novel analytical tools: tests of stationarity with respect to topology in the Bayesian estimation, and the use of coalescent simulations to test the significance of the population growth-rate parameter. PMID- 15545256 TI - Model selection and model averaging in phylogenetics: advantages of akaike information criterion and bayesian approaches over likelihood ratio tests. AB - Model selection is a topic of special relevance in molecular phylogenetics that affects many, if not all, stages of phylogenetic inference. Here we discuss some fundamental concepts and techniques of model selection in the context of phylogenetics. We start by reviewing different aspects of the selection of substitution models in phylogenetics from a theoretical, philosophical and practical point of view, and summarize this comparison in table format. We argue that the most commonly implemented model selection approach, the hierarchical likelihood ratio test, is not the optimal strategy for model selection in phylogenetics, and that approaches like the Akaike Information Criterion (AIC) and Bayesian methods offer important advantages. In particular, the latter two methods are able to simultaneously compare multiple nested or nonnested models, assess model selection uncertainty, and allow for the estimation of phylogenies and model parameters using all available models (model-averaged inference or multimodel inference). We also describe how the relative importance of the different parameters included in substitution models can be depicted. To illustrate some of these points, we have applied AIC-based model averaging to 37 mitochondrial DNA sequences from the subgenus Ohomopterus(genus Carabus) ground beetles described by Sota and Vogler (2001). PMID- 15545257 TI - Effects of tree shape on the accuracy of maximum likelihood-based ancestor reconstructions. PMID- 15545258 TI - The phylogenetic definition of reptilia. PMID- 15545259 TI - Metacanthomorpha: essay on a phylogeny-oriented database for morphology--the acanthomorph (teleostei) example. PMID- 15545262 TI - Sclerostin inhibition of Wnt-3a-induced C3H10T1/2 cell differentiation is indirect and mediated by bone morphogenetic proteins. AB - High bone mass diseases are caused both by activating mutations in the Wnt pathway and by loss of SOST, a bone morphogenetic protein (BMP) antagonist, leading to the activation of BMP signaling. Given the phenotypic similarity between mutations that activate these signaling pathways, it seems likely that BMPs and Wnts operate in parallel or represent components of the same pathway, modulating osteoblast differentiation. In this study, we show that in C3H10T1/2 cells, Wnt-3A and BMP-6 proteins were inducers of osteoblast differentiation, as measured by alkaline phosphatase (ALP) induction. Surprisingly, sclerostin, noggin, and human BMP receptor 1A (BMPR1A)-FC fusion proteins blocked Wnt-3A induced ALP as well as BMP-6-induced ALP activity. Dkk-1, a Wnt inhibitor, blocked Wnt-induced ALP activity but not BMP-induced ALP activity. Early Wnt-3A signaling as measured by beta-catenin accumulation was not affected by the BMP antagonists but was blocked by Dkk-1. Wnt-3A induced the appearance of BMP-4 mRNA 12 h prior to that of ALP in C3H10T1/2 cells. We propose that sclerostin and other BMP antagonists do not block Wnt signaling directly. Sclerostin blocks Wnt induced ALP activity by blocking the activity of BMP proteins produced by Wnt treatment. The expression of BMP proteins in this autocrine loop is essential for Wnt-3A-induced osteoblast differentiation. PMID- 15545263 TI - The biology of cyclic GMP-dependent protein kinases. PMID- 15545264 TI - Pseudomonas invasion of type I pneumocytes is dependent on the expression and phosphorylation of caveolin-2. AB - Pseudomonas aeruginosa is a major cause of pneumonia in patients with cystic fibrosis and other immuncompromising conditions. Here we showed that P. aeruginosa invades type I pneumocytes via a lipid raft-mediated mechanism. P. aeruginosa invasion of rat primary type I-like pneumocytes as well as a murine lung epithelial cell line 12 (MLE-12) is inhibited by drugs that remove membrane cholesterol and disrupt lipid rafts. Confocal microscopy demonstrated co localization of intracellular P. aeruginosa with lipid raft components including caveolin-1 and -2. We generated caveolin-1 and -2 knockdowns in MLE-12 cells by using RNA interference techniques. Decreased expression of caveolin-2 significantly impaired the ability of P. aeruginosa to invade MLE-12 cells. In addition, the lipid raft-dependent tyrosine phosphorylation of caveolin-2 appeared to be a critical regulator of P. aeruginosa invasion. PMID- 15545265 TI - Identification and characterization of the terminal enzyme of siroheme biosynthesis from Arabidopsis thaliana: a plastid-located sirohydrochlorin ferrochelatase containing a 2FE-2S center. AB - Higher plant sulfite and nitrite reductases contain siroheme as a prosthetic group. Siroheme is synthesized from the tetrapyrrole primogenitor uroporphyrinogen III in three steps involving methylation, oxidation, and ferrochelation reactions. In this paper we report on the Arabidopsis thaliana sirohydrochlorin ferrochelatase At-SirB. The complete precursor protein of 225 amino acids and shorter constructs in which the first 46 or 79 residues had been removed were shown to complement a defined Escherichia coli sirohydrochlorin ferrochelatase mutant. The mature form of the protein appeared to consist of only 150 amino acids, making it much smaller than previously characterized ferrochelatases. Green fluorescent protein tagging revealed that it is located in the chloroplast. The enzyme was easily produced in E. coli as a recombinant protein, and the isolated enzyme was found to have a specific activity of 48.5 nmol/min/mg. Significantly, the protein purified as a brown-colored solution with a UV-visible spectrum containing maxima at 415 and 455 nm, suggestive of an Fe-S center. EPR analysis of the recombinant protein produced a rhombic spectrum with G-values of 2.04, 1.94, and 1.90 and with temperature dependence consistent with a 2Fe-2S center. Redox titration demonstrated that the Fe-S center is highly unstable, with an apparent midpoint reduction potential of about -370 mV. This is the first Fe-S center to be reported in a higher plant ferrochelatase. The implications of the Fe-S center in an enzyme that is so closely associated with the metabolism of sulfur and iron are discussed. PMID- 15545266 TI - Crystal structures of the FXIa catalytic domain in complex with ecotin mutants reveal substrate-like interactions. AB - Thrombosis can lead to life-threatening conditions such as acute myocardial infarction, pulmonary embolism, and stroke. Although commonly used anti-coagulant drugs, such as low molecular weight heparin and warfarin, are effective, they carry a significant risk of inducing severe bleeding complications, and there is a need for safer drugs. Activated Factor XI (FXIa) is a key enzyme in the amplification phase of the coagulation cascade. Anti-human FXI antibody significantly reduces thrombus growth in a baboon thrombosis model without bleeding problems (Gruber, A., and Hanson, S. R. (2003) Blood 102, 953-955). Therefore, FXIa is a potential target for anti-thrombosis therapy. To determine the structure of FXIa, we derived a recombinant catalytic domain of FXI, consisting of residues 370-607 (rhFXI370-607). Here we report the first crystal structure of rhFXI370-607 in complex with a substitution mutant of ecotin, a panserine protease protein inhibitor secreted by Escherichia coli, to 2.2 A resolution. The presence of ecotin not only assisted in the crystallization of the enzyme but also revealed unique structural features in the active site of FXIa. Subsequently, the sequence from P5 to P2' in ecotin was mutated to the FXIa substrate sequence, and the structures of the rhFXI370-607-ecotin mutant complexes were determined. These structures provide us with an understanding of substrate binding interactions of FXIa, the structural information essential for the structure-based design of FXIa-selective inhibitors. PMID- 15545267 TI - Regulation of P-Rex1 by phosphatidylinositol (3,4,5)-trisphosphate and Gbetagamma subunits. AB - P-Rex1 is a guanine-nucleotide exchange factor (GEF) for the small GTPase Rac. We have investigated here the mechanisms of stimulation of P-Rex1 Rac-GEF activity by the lipid second messenger phosphatidylinositol (3,4,5)-trisphosphate (PtdIns(3,4,5)P3) and the Gbetagamma subunits of heterotrimeric G proteins. We show that a P-Rex1 mutant lacking the PH domain (DeltaPH) cannot be stimulated by PtdIns(3,4,5)P3, which implies that the PH domain confers PtdIns(3,4,5)P3 regulation of P-Rex1 Rac-GEF activity. Consistent with this, we found that PtdIns(3,4,5)P3 binds to the PH domain of P-Rex1 and that the DH/PH domain tandem is sufficient for PtdIns(3,4,5)P3-stimulated P-Rex1 activity. The Rac-GEF activities of the DeltaPH mutant and the DH/PH domain tandem can both be stimulated by Gbetagamma subunits, which infers that Gbetagamma subunits regulate P-Rex1 activity by binding to the catalytic DH domain. Deletion of the DEP, PDZ, or inositol polyphosphate 4-phosphatase homology domains has no major consequences on the abilities of either PtdIns(3,4,5)P3 or Gbetagamma subunits to stimulate P-Rex1 Rac-GEF activity. However, the presence of any of these domains impacts on the levels of basal and/or stimulated P-Rex1 Rac-GEF activity, suggesting that there are important functional interactions between the DH/PH domain tandem and the DEP, PDZ, and inositol polyphosphate 4-phosphatase homology domains of P-Rex1. PMID- 15545268 TI - HOXA5-twist interaction alters p53 homeostasis in breast cancer cells. AB - The homeotic gene HOXA5 has been shown to play an important role in breast tumorigenesis. We have shown that loss of p53 correlated with loss of a developmentally regulated transcription factor, HOXA5, in primary breast cancer. Searching for potential protein interacting partners we found that HOXA5 binds to an anti-apoptotic protein, Twist. Furthermore, Twist-overexpressing MCF-7 cells displayed a deregulated p53 response to gamma-radiation and decreased regulation of downstream target genes. Using a p53-promoter-reporter system, we demonstrated that HOXA5 could partially restore the inhibitory effects of Twist on p53 target genes. These effects are likely mediated through both the transcriptional up regulation of p53 and the protein-protein interaction between HOXA5 and Twist. Thus, the loss of HOXA5 expression could lead to the functional activation of Twist resulting in aberrant cell cycle regulation and promoting breast carcinogenesis. PMID- 15545269 TI - Akt1/Akt2 and mammalian target of rapamycin/Bim play critical roles in osteoclast differentiation and survival, respectively, whereas Akt is dispensable for cell survival in isolated osteoclast precursors. AB - Akt, also known as protein kinase B, is a serine/threonine protein kinase with antiapoptotic activities; also, it is a downstream target of phosphatidylinositol 3-kinase. Here we show that Akt1/Akt2 play a critical role in osteoclast differentiation but not cell survival and that mammalian target of rapamycin (mTOR) and Bim, a pro-apoptotic Bcl-2 family member, are required for cell survival in isolated osteoclast precursors. To investigate the function of Akt1, Akt2, mTOR, and Bim, we employed a retroviral system for delivery of small interfering RNA into cells. Loss of Akt1 and/or Akt2 protein inhibited osteoclast differentiation due to down-regulation of IkappaB-kinase (IKK) alpha/beta activity, phosphorylation of IkappaB-alpha, nuclear translocation of nuclear factor-kappaB (NFkappaB) p50, and NFkappaB p50 DNA-binding activity. Surprisingly, deletion of Akt1 and/or Akt2 protein did not stimulate cleaved caspase-3 activity and failed to promote apoptosis. Conversely, loss of mTOR protein induced apoptosis due to up-regulation of cleaved caspase-3 activity. In addition, we found that mTOR is downstream of phosphatidylinositol 3-kinase (but not Akt) and that macrophage colony-stimulating factor regulates Bim expression through mTOR activation for cell survival. These results demonstrate that Akt1/Akt2 are key elements in osteoclast differentiation and that the macrophage colony-stimulating factor stimulation of mTOR leading to Bim inhibition is essential for cell survival in isolated osteoclast precursors. PMID- 15545270 TI - Autoimmunity against a tissue kallikrein in IQI/Jic Mice: a model for Sjogren's syndrome. AB - We have recently characterized IQI/Jic mice as a model for Sjogren's syndrome (SS), a chronic autoimmune disease in humans. In SS, local lymphocytic infiltrations into salivary and lacrimal glands frequently develop to the involvement of systemic exocrine and nonexocrine organs, and the mechanism for progression of this disease remains obscure. Herein, we report identification of an autoantigen shared by various target organs in IQI/Jic mice. Polypeptides identified based on immunorecognition by autoantibodies in sera from IQI/Jic mice affected with autoimmune disease (>12 weeks of age) were tissue kallikrein (Klk) 1 and -13 and were cross-reactive to the autoantibodies. Interestingly, Klk-13, but not Klk-1, caused a proliferative response of splenic T cells from IQI/Jic mice from the age of 4 weeks onward. In addition, remarkably enhanced expression of Klk-13 was observed in the salivary glands of the mice in accordance with the development of inflammatory lesions. These results indicate that Klk-13 acts as an autoantigen and may increase T cells responsive to organs commonly expressing Klk-13, playing a pivotal role in the etiology of progression of disease in IQI/Jic mice. Our findings provide insights into the contributions of autoantigens shared by multiple organs in the progress of SS from an organ specific to a systemic disorder. PMID- 15545271 TI - Mechanical stretch stimulates protein kinase B/Akt phosphorylation in epidermal cells via angiotensin II type 1 receptor and epidermal growth factor receptor. AB - Mechanical stress is known to modulate fundamental events such as cell life and death. Mechanical stretch in particular has been identified as a positive regulator of proliferation in skin keratinocytes and other cell systems. In the present study it was investigated whether antiapoptotic signaling is also stimulated by mechanical stretch. It was demonstrated that mechanical stretch rapidly induced the phosphorylation of the proto-oncogene protein kinase B (PKB)/Akt at both phosphorylation sites (serine 473/threonine 308) in different epithelial cells (HaCaT, A-431, and human embryonic kidney-293). Blocking of phosphoinositide 3-OH kinase by selective inhibitors (LY-294002 and wortmannin) abrogated the stretch-induced PKB/Akt phosphorylation. Furthermore mechanical stretch stimulated phosphorylation of epidermal growth factor receptor (EGFR) and the formation of EGFR membrane clusters. Functional blocking of EGFR phosphorylation by either selective inhibitors (AG1478 and PD168393) or dominant negative expression suppressed stretch-induced PKB/Akt phosphorylation. Finally, the angiotensin II type 1 receptor (AT1-R) was shown to induce positive transactivation of EGFR in response to cell stretch. These findings define a novel signaling pathway of mechanical stretch, namely the activation of PKB/Akt by transactivation of EGFR via angiotensin II type 1 receptor. Evidence is provided that stretch-induced activation of PKB/Akt protects cells against induced apoptosis. PMID- 15545272 TI - Differential regulation of endoplasmic reticulum structure through VAP-Nir protein interaction. AB - The endoplasmic reticulum (ER) exhibits a characteristic tubular structure that is dynamically rearranged in response to specific physiological demands. However, the mechanisms by which the ER maintains its characteristic structure are largely unknown. Here we show that the integral ER-membrane protein VAP-B causes a striking rearrangement of the ER through interaction with the Nir2 and Nir3 proteins. We provide evidence that Nir (Nir1, Nir2, and Nir3)-VAP-B interactions are mediated through the conserved FFAT (two phenylalanines (FF) in acidic tract) motif present in Nir proteins. However, each interaction affects the structural integrity of the ER differently. Whereas the Nir2-VAP-B interaction induces the formation of stacked ER membrane arrays, the Nir3-VAP-B interaction leads to a gross remodeling of the ER and the bundling of thick microtubules along the altered ER membranes. In contrast, the Nir1-VAP-B interaction has no apparent effect on ER structure. We also show that the Nir2-VAP-B interaction attenuates protein export from the ER. These results demonstrate new mechanisms for the regulation of ER structure, all of which are mediated through interaction with an identical integral ER-membrane protein. PMID- 15545273 TI - The tandem BRCT domains of Ect2 are required for both negative and positive regulation of Ect2 in cytokinesis. AB - Epithelial cell transforming protein 2 (Ect2) is a guanine nucleotide exchange factor (GEF) for Rho GTPases, molecular switches essential for the control of cytokinesis in mammalian cells. Aside from the canonical Dbl homology/pleckstrin homology cassette found in virtually all Dbl family members, Ect2 contains N terminal tandem BRCT domains. In this study, we address the role of the Ect2 BRCT domains in the regulation of Ect2 activity and cytokinesis. First, we show that the depletion of endogenous Ect2 by small interfering RNA induces multinucleation, suggesting that Ect2 is required for cytokinesis. In addition, we provide evidence that Ect2 normally exists in an inactive conformation, which is at least partially due to an intramolecular interaction between the BRCT domains and the C-terminal domain of Ect2. This intramolecular interaction masks the catalytic domain responsible for guanine nucleotide exchange toward RhoA. Consistent with a role in regulating Ect2 GEF activity, overexpression of an N terminal Ect2 containing the tandem BRCT domains, but not single BRCT domain or BRCT domain mutant, leads to a failure in cytokinesis. Surprisingly, although ectopically expressed wild-type Ect2 rescues the multinucleation resulting from the depletion of endogenous Ect2, expression of a BRCT mutant of Ect2 failed to restore proper cytokinesis in these cells. Taken together, the results of our study indicate that the tandem BRCT domains of Ect2 play dual roles in the regulation of Ect2. Whereas these domains negatively regulate Ect2 GEF activity in interphase cells, they are also required for the proper function of Ect2 during cytokinesis. PMID- 15545274 TI - Modeling of Galpha(s) and Galpha(i) regulation of human type V and VI adenylyl cyclase. AB - We examined the kinetics of Galpha(s) and Galpha(i) regulation of human type V and type VI adenylyl cyclase (AC V and AC VI) in order to better model interactions between AC and its regulators. Activation of AC VI by Galpha(s) displayed classical Michaelis-Menten kinetics, whereas AC V activation by Galpha(s) was cooperative with a Hill coefficient of 1.4. The basal activity of human AC V, but not that of AC VI, was inhibited by Galpha(i). Both enzymes showed greater inhibition by Galpha(i) at low Galpha(s) concentrations; however, human AC V was activated by Galpha(i) at high Galpha(s) concentrations. Neither regulator had an effect on the K(m) for Mg-ATP. Mutations made within the Galpha(s) binding pocket of AC V (N1090D) and VI (F1078S) displayed 6- and 14 fold greater EC(50) values for Galpha(s) activation but had no effect on Galpha(i) inhibition of basal activity or K(m) for Mg-ATP. Galpha(s)-stimulated AC VI-F1078S was not significantly inhibited by Galpha(i), despite normal inhibition by Galpha(i) upon forskolin stimulation. Mechanistic models for Galpha(s) and Galpha(i) regulation of AC V and VI were derived to describe these results. Our models are consistent with previous studies, predicting a decrease in affinity of Galpha(i) in the presence of Galpha(s). For AC VI, Galpha(s) is required for inhibition but not binding by Galpha(i). For AC V, binding of two molecules of Galpha(s) and Galpha(i) to an AC dimer are required to fully describe the data. These models highlight the differences between AC V and VI and the complex interactions with two important regulators. PMID- 15545275 TI - Early growth response-1 regulates lipopolysaccharide-induced suppressor of cytokine signaling-1 transcription. AB - Suppressor of cytokine signaling (SOCS)-1 is a critical regulator of lipopolysaccharide (LPS) tolerance and LPS-induced cytokine production. The mechanisms regulating the transcription of SOCS-1 in response to LPS are not entirely understood. Functional analysis of the SOCS-1 promoter demonstrates that early growth response-1 (Egr-1) is an important transcriptional regulator of SOCS 1. Two Egr-1 binding sites are present within the SOCS-1 promoter as shown by EMSA and supershift analysis. Further, mutation of the Egr-1 binding sites significantly reduces both the basal and LPS-induced transcriptional activity of the promoter. Chromatin immunoprecipitation experiments confirm LPS-induced binding of Egr-1 to the SOCS-1 promoter in vivo. Additionally, Egr-1(-/-) macrophages show reduced levels of LPS-induced SOCS-1 expression in comparison with macrophages derived from Egr-1(+/+) littermate controls. These results demonstrate an important role for Egr-1 in regulating both the basal and LPS induced activity of the SOCS-1 promoter. PMID- 15545276 TI - Rom2p, the Rho1 GTP/GDP exchange factor of Saccharomyces cerevisiae, can mediate stress responses via the Ras-cAMP pathway. AB - The Ras-cyclic AMP pathway is connected to other nutrient-regulated signaling pathways and mediates the global stress responses of Saccharomyces cerevisiae. Here, we show that Rom2p, the Rho1 GTP/GDP exchange factor, can mediate stress responses and cell growth via the Ras-cAMP pathways. ROM2 was isolated as a suppresser of heat and NaCl sensitivity caused by the lack of the Ras-GTPase activator Ira2p or of cAMP phosphodiesterases. Subsequent analysis of strains with a rom2 deletion showed that Rom2p is essential for resistance to a variety of stresses caused by freeze-thawing, oxidants, cycloheximide, NaCl, or cobalt ions. Stress sensitivity and the growth defect caused by the rom2 deletion could be suppressed by depleting Ras or protein kinase A (PKA) activity or by overexpressing the high affinity cAMP phosphodiesterase Pde2p. In addition, overexpression of ROM2 could not rescue cells lacking the regulatory subunit of PKA, indicating that the Ras-adenylate, cyclase-PKA cascade is essential for Rom2p-mediated stress responses and cell growth. Deletion of IRA2 exacerbated the freeze-thaw sensitivity and growth defect of the rom2 mutant, indicating that Rom2p signaling may control Ras independently of IRA2. Increases in cAMP levels were detected in the rom2 deletion mutants, and these were comparable with the effects of an ira2 mutation. The effects of the deletion of ROM2 on sensitivity to hydrogen peroxide, paraquat, and cobalt ions, but not to caffeine, were reduced when a constitutive allele of RHO1 was introduced on a single copy plasmid. However, the effects of the deletion of ROM2 on sensitivity to diamide and NaCl were exacerbated. Taken together, our data indicate that Rom2p can regulate PKA activity by controlling cAMP levels via the Ras-cAMP pathway and that for those stresses related to oxidative stress, this cross-talk is probably mediated via the Rho1p-activated MAPK pathway. PMID- 15545277 TI - A temperature-sensitive mechanism that regulates post-translational stability of a plastidial omega-3 fatty acid desaturase (FAD8) in Arabidopsis leaf tissues. AB - Trienoic fatty acids (TAs) are the major constituents in plant membrane lipids. In Arabidopsis, two plastidial isozymes of omega-3 fatty acid desaturase, FAD7 and FAD8, are the major contributors for TA production in leaf tissues. Despite a high degree of structural relatedness, activities of these two isozymes are regulated differentially in response to temperature. Elevated temperatures lead to decreases in leaf TA level due to temperature sensitivity of FAD8 activity. A series of FAD7-FAD8 chimeric genes, each encoding a functional plastidial omega-3 desaturase, were introduced into the Arabidopsis fad7fad8 double mutant. Constructs with or without a c-Myc epitope tag were tested. Functionality of each chimeric gene in response to temperature was assayed by Northern and Western analyses and by examining the fatty acid composition. All transformants harboring a chimeric gene containing the FAD8-derived C-terminal coding region (44 amino acids) showed a marked decrease in TA level when exposed to high temperature, similarly as transgenic lines complemented with the native form of FAD8. The reduction of TA level was accompanied by a decrease in the amount of omega-3 desaturase protein but not necessarily by a decrease in its transcript level. Analysis of the decay of c-Myc-tagged products after inhibiting protein synthesis revealed that the FAD8-derived C-terminal region acts in an autoregulatory fashion to destabilize the protein at high temperature. This suggests that the regulation of post-translational stability of FAD8 provides an important regulatory mechanism for modifying its activity in response to temperature, mediating a decrease in TA level at elevated temperatures. PMID- 15545278 TI - Spatial integration of TIP47 and adipophilin in macrophage lipid bodies. AB - We studied the distribution of the PAT family proteins TIP47 and adipophilin in lipid bodies of THP-1 cell-derived macrophages using freeze-fracture immunolabeling and other techniques. Lipid bodies in macrophages comprise lipid droplets and extensive, previously scantily characterized sheet-like organelles, which we descriptively call "lipid sails." TIP47 and adipophilin are components of many, but not all, the lipid droplets. Both proteins are not confined to the surface of lipid droplets, as supposed, but are also inside lipid droplet cores. They are not codistributed stoichiometrically in lipid droplets. How TIP47 and adipophilin, which are polar proteins, enter the lipid droplets and are packaged among the hydrophobic neutral lipids of the core is unclear. However, in the lipid layers of the core, these proteins are directed sometimes inward and sometimes outward. Because TIP47 and adipophilin also localize to lipid sails, lipid sails are intimately involved in intracellular lipid metabolism. PMID- 15545279 TI - A dual role for the N-terminal region of Mycobacterium tuberculosis Hsp16.3 in self-oligomerization and binding denaturing substrate proteins. AB - The N-terminal regions, which are highly variable in small heat-shock proteins, were found to be structurally disordered in all the 24 subunits of Methanococcus jannaschii Hsp16.5 oligomer and half of the 12 subunits of wheat Hsp16.9 oligomer. The structural and functional roles of the corresponding region (potentially disordered) in Mycobacterium tuberculosis Hsp16.3, existing as nonamers, were investigated in this work. The data demonstrate that the mutant Hsp16.3 protein with 35 N-terminal residues removed (DeltaN35) existed as trimers/dimers rather than as nonamers, failing to bind the hydrophobic probe (1,1'-bi(4-anilino)naphthalene-5,5'-disulfonic acid) and exhibiting no chaperone like activity. Nevertheless, another mutant protein with the C-terminal extension (of nine residues) removed, although existing predominantly as dimers, exhibited efficient chaperone-like activity even at room temperatures, indicating that pre existence as nonamers is not a prerequisite for its chaperone-like activity. Meanwhile, the mutant protein with both the N- and C-terminal ends removed fully exists as a dimer lacking any chaperone-like activity. Furthermore, the N terminal region alone, either as a synthesized peptide or in fusion protein with glutathione S-transferase, was capable of interacting with denaturing proteins. These observations strongly suggest that the N-terminal region of Hsp16.3 is not only involved in self-oligomerization but also contains the critical site for substrate binding. Such a dual role for the N-terminal region would provide an effective mechanism for the small heat-shock protein to modulate its chaperone like activity through oligomeric dissociation/reassociation. In addition, this study demonstrated that the wild-type protein was able to form heterononamers with DeltaN35 via subunit exchange at a subunit ratio of 2:1. This implies that the 35 N-terminal residues in three of the nine subunits in the wild-type nonamer are not needed for the assembly of nonamers from trimers and are thus probably structurally disordered. PMID- 15545280 TI - Contribution of N-linked glycans to the conformation and function of intercellular adhesion molecules (ICAMs). AB - The crystal structures of the glycosylated N-terminal two domains of ICAM-1 and ICAM-2 provided a framework for understanding the role of glycosylation in the structure and function of intercellular adhesion molecules (ICAMs). The most conserved glycans were less flexible in the structures, interacting with protein residues and contributing to receptor folding and expression. The first N-linked glycan in ICAM-2 contacts an exposed tryptophan residue, defining a conserved glycan-W motif critical for the conformation of the integrin binding domain. The absence of this motif in human ICAM-1 exposes regions used in receptor dimerization and rhinovirus recognition. Experiments with soluble molecules having the N-terminal two domains of human ICAMs identified glycans of the high mannose type N-linked to the second domain of the dendritic cell-specific ICAM grabbing nonintegrin lectin-ligands ICAM-2 and ICAM-3. About 40% of those receptor molecules bear endoglycosidase H sensitive glycans responsible of the lectin binding activity. High mannose glycans were absent in ICAM-1, which did not bind to the lectin, but they appeared in ICAM-1 mutants with additional N linked glycosylation and lectin binding activity. N-Linked glycosylation regulate both conformation and immune related functions of ICAM receptors. PMID- 15545281 TI - Taurine monochloramine activates a cell death pathway involving Bax and Caspase 9. AB - Taurine is an abundant free amino acid that interacts with the potent oxidant hypochlorous acid to form the less toxic and more stable oxidant taurine monochloramine (TauNHCl). TauNHCl has diverse cellular effects ranging from inhibiting the production of proinflammatory mediators to inhibiting cell proliferation and inducing cell death. We hypothesized that TauNHCl could activate a cell death pathway involving Bcl-2 members and the activation of caspase proteases. FL5.12 cells are lymphocytic cells that undergo apoptosis following interleukin-3 (IL-3) withdrawal. Therefore, cell death following TauNHCl treatment of FL5.12 cells was compared and contrasted with IL-3 withdrawal. We found that TauNHCl treatment activates a cell death pathway with kinetics very similar to IL-3 withdrawal. TauNHCl-treated cells undergo an annexin V-positive/propidium iodide-negative phase of death consistent with apoptosis. TauNHCl treatment results in a conformational change in BAX that is associated with its activation. Both Bcl-2 and, to a lesser degree, the dominant negative form of caspase-9 inhibit cell death following TauNHCl treatment. In contrast with IL-3 withdrawal, TauNHCl treatment of FL5.12 cells results in a rapid cell cycle arrest that is cell cycle phase-independent. These results demonstrate that TauNHCl treatment induces a rapid, cell cycle-independent proliferative arrest followed by the activation of a cell death pathway involving Bcl-2 family members and caspase activation. PMID- 15545282 TI - Estrogen-induced spermatogenic cell apoptosis occurs via the mitochondrial pathway: role of superoxide and nitric oxide. AB - The detrimental effects of estrogen on testicular function provide a conceptual basis to examine the speculative link between increased exposure to estrogens and spermatogenic cell death. Using an in vitro model, we provide an understanding of the events leading to estrogen-induced apoptosis in cells of spermatogenic lineage. Early events associated with estrogen exposure were up-regulation of FasL and increased generation of H(2)O(2), superoxide, and nitric oxide. The ability of anti-FasL antibodies to prevent several downstream biochemical changes and cell death induced by 17beta-estradiol substantiates the involvement of the cell death receptor pathway. Evidence for the amplification of the death-inducing signals through mitochondria was obtained from the transient mitochondrial hyperpolarization observed after estradiol exposure resulting in cytochrome c release. A combination of nitric oxide and superoxide but not H(2)O(2) was responsible for the mitochondrial hyperpolarization. Mn(III) tetrakis(4-benzoic acid)porphyrin chloride, an intracellular peroxynitrite scavenger, was able to reduce mitochondrial hyperpolarization and cell death. Although nitric oxide augmentation occurred through an increase in the expression of inducible nitric oxide synthase, superoxide up-regulation was a product of estradiol metabolism. All of the above changes were mediated through an estrogen receptor-based mechanism because tamoxifen, the estrogen receptor modulator, was able to rescue the cells from estrogen-induced alterations. This study establishes the importance of the independent capability of cells of the spermatogenic lineage to respond to estrogens and most importantly suggests that low dose estrogens can potentially cause severe spermatogenic cellular dysfunction leading to impaired fertility even without interference of the hypothalamo-hypophyseal axis. PMID- 15545283 TI - Ectopic expression of p73alpha, but not p73beta, suppresses myogenic differentiation. AB - The TRP73 gene, a member of the p53 family, encodes several variants through differential splicing and use of alternative promoters. At the N terminus, two different promoters generate the full-length and the DeltaN isoforms, with or without the transactivating domain. At the C terminus, seven isoforms generated through alternative splicing have been cloned. Previous studies have demonstrated that DeltaN-p73 interferes with p73-induced apoptosis. However, there has been no evidence for functional diversity of the C-terminal p73 variants. In this study, we found that p73alpha and p73beta exerted differential effect on the differentiation of C2C12 myoblasts. Although p73beta lacked any detectable effect on differentiation, p73alpha caused a substantial delay in the expression of muscle-specific genes. In co-transfection experiments p73alpha, but not p73beta, attenuated the transcriptional activity of MyoD. Microarray-based gene profiling confirmed the protraction of MyoD-dependent gene expression in C2C12 cells stably expressing p73alpha. Notwithstanding the differential effect on differentiation, p73alpha and p73beta showed similar activity in sensitizing C2C12 myoblasts to cisplatin-induced cell death. These results demonstrated a functional diversity between the two C-terminal variants of p73 and suggested that p73alpha can regulate cellular differentiation in addition to its role in stimulating cell death. PMID- 15545284 TI - p116Rip decreases myosin II phosphorylation by activating myosin light chain phosphatase and by inactivating RhoA. AB - p116Rip was originally found to be a RhoA-binding protein, but its function has been unknown. Here, we clarify the function of p116Rip. Two critical findings were made. First, we found that p116Rip activated the GTPase activity of RhoA in vitro and that p116Rip overexpression in cells consistently diminished the epidermal growth factor-induced increase in GTP-bound RhoA. Second, p116Rip activated the myosin light chain phosphatase (MLCP) activity of the holoenzyme. p116Rip did not activate the catalytic subunit alone, indicating that the activation is due to the binding of p116Rip to the myosin phosphatase targeting subunit MYPT1. Interestingly, the activation of phosphatase was specific to myosin as substrate, and p116Rip directly bound to myosin, thus facilitating myosin/MLCP interaction. The gene silencing of p116Rip consistently and significantly increased myosin phosphorylation as well as stress fiber formation in cells. Based upon these findings, we propose that p116Rip is an important regulatory component that controls the RhoA signaling pathway, thus regulating MLCP activity and myosin phosphorylation in cells. PMID- 15545285 TI - Actin activation of myosin heavy chain kinase A in Dictyostelium: a biochemical mechanism for the spatial regulation of myosin II filament disassembly. AB - Studies in Dictyostelium discoideum have established that the cycle of myosin II bipolar filament assembly and disassembly controls the temporal and spatial localization of myosin II during critical cellular processes, such as cytokinesis and cell locomotion. Myosin heavy chain kinase A (MHCK A) is a key enzyme regulating myosin II filament disassembly through myosin heavy chain phosphorylation in Dictyostelium. Under various cellular conditions, MHCK A is recruited to actin-rich cortical sites and is preferentially enriched at sites of pseudopod formation, and thus MHCK A is proposed to play a role in regulating localized disassembly of myosin II filaments in the cell. MHCK A possesses an aminoterminal coiled-coil domain that participates in the oligomerization, cellular localization, and actin binding activities of the kinase. In the current study, we show that the interaction between the coiled-coil domain of MHCK A and filamentous actin leads to an approximately 40-fold increase in the initial rate of kinase catalytic activity. Actin-mediated activation of MHCK A involves increased rates of kinase autophosphorylation and requires the presence of the coiled-coil domain. Structure-function analyses revealed that the coiled-coil domain alone binds to actin filaments (apparent K(D) = 0.9 microm) and thus mediates the direct interaction with F-actin required for MHCK A activation. Collectively, these results indicate that MHCK A recruitment to actin-rich sites could lead to localized activation of the kinase via direct interaction with actin filaments, and thus this mode of kinase regulation may represent an important mechanism by which the cell achieves localized disassembly of myosin II filaments required for specific changes in cell shape. PMID- 15545286 TI - Fall-related risk factors and osteoporosis in older women referred to an open access bone densitometry service. AB - OBJECTIVE: Both falls and low bone density are important in the pathogenesis of osteoporotic fractures. Whilst bone density is routinely measured to assess fracture risk, little attention is given to the assessment of fall risk. In this study we have determined the prevalence and explored relationships between fall related risk factors and osteoporosis in women referred to our open access bone densitometry service. DESIGN: Cross-sectional study. SETTING: Teaching hospital in south-west London, UK. SUBJECTS: Older women referred for open access bone densitometry. MEASUREMENTS: Bone densitometry by dual-energy X-ray absorptiometry and fall risk assessment (visual acuity, ability to do five stand-ups without arm use and ability to perform heel-toe walking). RESULTS: Data for 558 women seen over an 18 month period were examined. Their mean age was 74.8 years (range 65 93). Fall risk and femoral neck (FN) osteoporosis increased with age, with fall related risk factors being more prevalent than FN osteoporosis at each tertile of age. Women with both FN osteoporosis and fall-related risk factors ranged from 7% in the youngest tertile to 22% in the oldest tertile. In women with FN osteoporosis, increased fall risk was found in 37% in the youngest tertile, increasing to 63% in the oldest tertile. CONCLUSIONS: Fall-related risk factors are common in older women referred for open access bone densitometry. We recommend that both bone density and fall risk assessment, using simple screening tests for falls, are essential to determine fracture risk in older people referred for bone densitometry. Subsequent management to reduce fracture risk should be individualised for each patient. PMID- 15545288 TI - Bradykinin specificity and signaling at GPR100 and B2 kinin receptors. PMID- 15545287 TI - Identification of R(-)-isomer of efonidipine as a selective blocker of T-type Ca2+ channels. AB - Efonidipine, a derivative of dihydropyridine Ca(2+) antagonist, is known to block both L- and T-type Ca(2+) channels. It remains to be clarified, however, whether efonidipine affects other voltage-dependent Ca(2+) channel subtypes such as N-, P/Q- and R-types, and whether the optical isomers of efonidipine have different selectivities in blocking these Ca(2+) channels, including L- and T-types. To address these issues, the effects of efonidipine and its R(-)- and S(+)-isomers on these Ca(2+) channel subtypes were examined electrophysiologically in the expression systems using Xenopus oocytes and baby hamster kidney cells (BHK tk ts13). Efonidipine, a mixture of R(-)- and S(+)-isomers, exerted blocking actions on L- and T-types, but no effects on N-, P/Q- and R-type Ca(2+) channels. The selective blocking actions on L- and T-type channels were reproduced by the S(+) efonidipine isomer. By contrast, the R(-)-efonidipine isomer preferentially blocked T-type channels. The blocking actions of efonidipine and its enantiomers were dependent on holding potentials. These findings indicate that the R(-) isomer of efonidipine is a specific blocker of the T-type Ca(2+) channel. PMID- 15545289 TI - Bradykinin B2 and GPR100 receptors: a paradigm for receptor signal transduction pharmacology. AB - The aim of the present report was to investigate the ligand selectivity of the human orphan G-protein-coupled receptor GPR100 (hGPR100), recently identified as a novel bradykinin (BK) receptor, as compared with that of the human B(2) receptor (hB(2)R) stably transfected in Chinese hamster ovary cells. BK was able to inhibit the cAMP production induced by forskolin with a potency 100-fold lower at the hGPR100 (pEC(50) = 6.6) than that measured at the hB(2)R (pEC(50) = 8.6). Both effects were inhibited by the B(2) receptor antagonist Icatibant (1 microM). The nonpeptide B(2) receptor agonist FR190997 (8-[2,6-dichloro-3-[N methylcarbamoyl)cinnamidoacetyl]-N-methylamino]benzyloxy]-2-methyl-4-(2 pyridylmethoxy)quinoline) did inhibit the forskolin-induced cAMP production (pEC(50) = 7.7) at the hB(2)R, whereas it was not able to exert any effect at the hGPR100. The human insulin-like peptide relaxin 3 did inhibit the cAMP production at the hGPR100 (pEC(50) = 7.3) at a greater extent than BK, and was devoid of any effect at the hB(2)R. FR190997 and relaxin 3 responses at the hB(2)R and hGPR100, respectively, were not inhibited by Icatibant (1 microM). These data indicate FR190997 and relaxin 3 as selective agonists for hB(2)R and hGPR100, respectively, and support the concept that different agonists may specifically bias the conformational states of a receptor to result in a final common G protein coupling, which is differentially recognized by antagonists. PMID- 15545290 TI - Locomotor effects of imidazoline I2-site-specific ligands and monoamine oxidase inhibitors in rats with a unilateral 6-hydroxydopamine lesion of the nigrostriatal pathway. AB - The present study examined the ability of the selective imidazoline I(2)-site ligands 2-(-2-benzofuranyl)-2-imidazoline (2-BFI) and 2-[4,5-dihydroimidaz-2-yl] quinoline (BU224) and selected monoamine oxidase (MAO) inhibitors to evoke locomotor activity in rats bearing a lesion of the nigrostriatal pathway. Male Sprague-Dawley rats were injected with 12.5 microg 6-hydroxydopamine (6-OHDA) into the right median forebrain bundle to induce a unilateral lesion of the nigrostriatal tract. After 6 weeks, test drugs were administered either alone or in combination with L-DOPA (l-3,4-dihydroxyphenylamine) and the circling behaviour of animals was monitored as an index of anti-Parkinsonian activity. Intraperitoneal (i.p.) administration of the irreversible MAO-B inhibitor deprenyl (20 mg kg(-1)) or the imidazoline I(2)-site ligands BU224 (14 mg kg(-1)) and 2-BFI (7 and 14 mg kg(-1)) produced significant increases in ipsiversive rotations compared to vehicle controls totaling, at the highest respective doses tested, 521 +/-120, 131 +/- 37 and 92.5 +/- 16.3 net contraversive rotations in 30 (deprenyl) or 60 (BU224 and 2-BFI) min. In contrast, the reversible MAO-A inhibitor moclobemide (2.5-10 mg kg(-1)) and the reversible MAO-B inhibitor lazabemide (2.5-10 mg kg(-1)) failed to instigate significant rotational behaviour compared to vehicle. Coadministration of lazabemide (10 mg kg(-1)), moclobemide (10 mg kg(-1)) or 2-BFI (14 mg kg(-1)) with L-DOPA (20 mg kg(-1)) significantly increased either the duration or total number of contraversive rotations emitted over the testing period in comparison to L-DOPA alone. These data suggest that I(2)-specific ligands have dual effects in the 6-OHDA-lesioned rat model of Parkinson's disease; a first effect associated with an increase in activity in the intact hemisphere, probably via an increase in striatal dopamine content, and a secondary action which, through the previously documented inhibition of MAO-A and/or MAO-B, increases the availability of dopamine produced by L-DOPA. PMID- 15545291 TI - Cardiac Na+-Ca2+ exchanger current induced by tyrphostin tyrosine kinase inhibitors. AB - Tyrosine kinase (TK) inhibitors genistein and tyrphostin A23 (A23) inhibited Ca(2+) currents in guinea-pig ventricular myocytes investigated under standard whole-cell conditions (K(+)-free Tyrode's superfusate; EGTA-buffered (pCa-10.5) Cs(+) dialysate). However, the inhibitors (100 microM) also induced membrane currents that reversed between -40 and 0 mV, and the objective of the present study was to characterize these currents. Genistein-induced current behaved like Cl(-) current, and was unaffected by either the addition of divalent cations (0.5 mM Cd(2+); 3 mM Ni(2+)) that block the Na(+)-Ca(2+) exchanger (NCX), or the removal of external Na(+) and Ca(2+). A23-induced current was independent of Cl( ) driving force, and strongly suppressed by addition of Cd(2+) and Ni(2+), and by removal of either external Na(+) or Ca(2+). These and other results suggested that A23 activated an NCX current driven by submembrane Na(+) and Ca(2+) concentrations higher than those in the bulk cytoplasm. Improved control of intracellular Na(+) and Ca(2+) concentrations was obtained by suppressing cation influx (10 microM verapamil) and raising dialysate Na(+) to 7 mM and dialysate pCa to 7. Under these conditions, stimulation by A23 was described by the Hill equation with EC(50) 68 +/- 4 microM and coefficient 1.1, tyrphostin A25 was as effective as A23, and TK-inactive tyrphostin A1 was ineffective. Phosphotyrosyl phosphatase inhibitor orthovanadate (1 mM) antagonized the action of 100 microM A23. The results suggest that activation of cardiac NCX by A23 is due to inhibition of genistein-insensitive TK. PMID- 15545292 TI - Regulation of cardiac Na-Ca exchange activity by selective tyrosine kinase inhibition. PMID- 15545293 TI - Population based randomised controlled trial on impact of screening on mortality from abdominal aortic aneurysm. AB - OBJECTIVE: To assess whether screening for abdominal aortic aneurysms in men reduces mortality. DESIGN: Population based randomised controlled trial of ultrasound screening, with intention to treat analysis of age standardised mortality. SETTING: Community based screening programme in Western Australia. PARTICIPANTS: 41,000 men aged 65-83 years randomised to intervention and control groups. INTERVENTION: Invitation to ultrasound screening. MAIN OUTCOME MEASURE: Deaths from abdominal aortic aneurysm in the five years after the start of screening. RESULTS: The corrected response to invitation to screening was 70%. The crude prevalence was 7.2% for aortic diameter > or = 30 mm and 0.5% for diameter > or = 55 mm. Twice as many men in the intervention group than in the control group underwent elective surgery for abdominal aortic aneurysm (107 v 54, P = 0.002, chi2 test). Between scheduled screening and the end of follow up 18 men in the intervention group and 25 in the control group died from abdominal aortic aneurysm, yielding a mortality ratio of 0.61 (95% confidence interval 0.33 to 1.11). Any benefit was almost entirely in men aged between 65 and 75 years, where the ratio was reduced to 0.19 (0.04 to 0.89). CONCLUSIONS: At a whole population level screening for abdominal aortic aneurysms was not effective in men aged 65-83 years and did not reduce overall death rates. The success of screening depends on choice of target age group and the exclusion of ineligible men. It is also important to assess the current rate of elective surgery for abdominal aortic aneurysm as in some communities this may already approach a level that reduces the potential benefit of population based screening. PMID- 15545294 TI - Induction of a sunflower CC-NBS-LRR resistance gene analogue during incompatible interaction with Plasmopara halstedii. AB - Downy mildew caused by Plasmopara halstedii is one of the main diseases causing economic losses in cultivated sunflower. Resistance in this host is conferred by major genes denoted Pl. The inbred sunflower line QIR8, which contains the Pl8 locus and is resistant to all known downy mildew races, was used to isolate a full-length resistance gene analogue (RGA) belonging to the CC-NBC-LRR class of plant resistance genes. The genetically incompatible combination involving downy mildew races 300 and sunflower line QIR8 was characterized by a hypersensitive like reaction. Semi-quantitative RT-PCR analysis showed that the transcript of Ha NTIR11g RGA was specifically induced during the incompatible reaction. The transcript was induced approximately 3 d post-infection (dpi), and then decreased by 9 dpi. The high level of transcriptional expression of this RGA coincides with a transcript accumulation of the hsr203J gene which is a marker of the hypersensitive reaction. Treatment with signalling molecules, including salicylic acid and methyl jasmonate, did not activate transcription of the Ha-NTIR11g gene, indicating that Ha-NTIR11g expression is not regulated by defence signalling pathways triggered by these molecules. Ha-NTIR11g was not induced by treatment with hydrogen peroxide or wounding. These results suggest that Ha-NTIR11g RGA may play a critical role in protecting sunflower cells against P. halstedii. The transcript accumulation of R gene-mediated signalling components was also examined. PMID- 15545295 TI - Differential response of PCNA and Cdk-A proteins and associated kinase activities to benzyladenine and abscisic acid during maize seed germination. AB - The proliferating cell nuclear antigen (PCNA) is a protein factor required for processive DNA synthesis that is associated with G(1) cell cycle proteins. It has been demonstrated previously that, in germinating maize (Zea mays) embryonic axes, PCNA forms protein complexes with two Cdk-A proteins (32 and 36 kDa) and with a putative D-type cyclin. These complexes exhibit protein kinase activity on histone H1 and on the maize homologue of the pRB (retinoblastoma) protein. Flow cytometry has been used to study the influence of the phytohormones benzyladenine (BA) and abscisic acid (ABA) on cell cycle advancement during maize germination. It was found that, while BA accelerates the passage of cells from G(1) to G(2), ABA delays cell cycle events so that most cells seem to remain in G(1). The amounts of PCNA and Cdk-A proteins also vary according to the hormone treatment. In embryonic axes, PCNA increases rapidly during early germination in BA, compared with a gradual increase in water, while ABA treatment had only a marginal effect. However, of the two Cdk-A proteins, the 32 kDa protein is strongly reduced after 15 h of imbibition in water while this occurs later when axes are imbibed in BA or ABA. The PCNA-associated protein kinase activity in the BA and ABA treatments falls after 3 h of imbibition compared with activity in the control; however, while kinase activity in the BA treatment continues to decline during imbibition, it remains relatively constant until 24 h of imbibition in the ABA treatment. By contrast, a p13(Suc1)-associated Cdk-A kinase is activated after 15 h of imbibition under all treatments, particularly in ABA. These results suggest that, in maize, ABA delays the germination process by affecting cell cycle advancement, stopping cells mostly in a G(1) state. PMID- 15545296 TI - FtsH-mediated repair of the photosystem II complex in response to light stress. AB - A common feature of light stress in plants, algae, and cyanobacteria is the light induced damage to the photosystem II complex (PSII), which catalyses the photosynthetic oxidation of water to molecular oxygen. A repair cycle operates to replace damaged subunits within PSII, in particular, the D1 reaction centre polypeptide, by newly synthesized copies. As yet the molecular details of this physiologically important process remain obscure. A key aspect of the process that has attracted much attention is the identity of the protease or proteases involved in D1 degradation. The results are summarized here of recent mutagenesis experiments that were designed to assess the functional importance of the DegP/HtrA and FtsH protease families in the cyanobacterium Synechocystis sp. PCC 6803. Based on these results and the analysis of Arabidopsis mutants, a general model for PSII repair is suggested in which FtsH complexes alone are able to degrade damaged D1. PMID- 15545297 TI - Enzymes, metabolites and fluxes. AB - Aspects of metabolic control theory and experiments from metabolic biochemistry are reviewed in order to deduce the circumstances in which experimental studies involving metabolomics have the greatest chance of success. It is concluded that metabolic changes effected mainly through a single enzyme are those most likely to lead to large changes in metabolite concentrations. Metabolic changes brought about through signal transduction mechanisms will tend to result in relatively much smaller adjustments in metabolite concentrations, whilst allowing significant changes in metabolic rates. PMID- 15545298 TI - Circadian regulation of cell cycle and apoptosis proteins in mouse bone marrow and tumor. AB - Proapoptotic drugs such as docetaxel displayed least toxicity and highest antitumor efficacy following dosing during the circadian rest phase in mice, suggesting that cell cycle and apoptotic processes could be regulated by the circadian clock. In study 1, mouse bone marrow and/or tumor were obtained every 4 h for 24 h in C3H/HeN mice with or without MA13/C mammary adenocarcinoma in order to determine the circadian patterns in cell-cycle phase distribution and BCL-2 anti-apoptotic protein expression. In study 2, mouse bone marrow from B6D2F1 mice was sampled every 3 h for 24 h in order to confirm the BCL-2 rhythm and to study its relation with 24 h changes in the expression of proapoptotic BCL-2-associated X protein (BAX) protein and clock genes mPer2, mBmal1, mClock, and mTim mRNAs. The rhythms in G1-, S- or G2/M-phase cells were shifted in tumor compared with bone marrow. In the tumor, the mean proportion of G2/M-phase cells increased by 75% from late rest to late activity span (P from cosinor = 0.001). No 24 h rhythm was found for BCL-2 in tumors. In contrast to this, in the bone marrow, mean BCL 2 expression varied 2.8-fold in B6D2F1 mice (P=0.025) and 3- or 4.5-fold in tumor bearing and nontumor-bearing C3H/HeN mice, with a peak during the early rest span (P=0.024 and P<0.001, respectively). BAX varied fivefold during the 24 h span with a major peak occurring near mid-activity (P=0.007). The mean mRNAs of mPer2, mClock, and mBmal1 varied twofold to threefold over the 24 h, with high values during the activity span (P<0.05). In the tumor, the circadian organization in cell-cycle phase distribution was shifted and BCL2 rhythm was ablated. Conversely, a molecular circadian clock likely regulated BCL-2 and BAX expression in the bone marrow, increasing cellular protection against apoptosis during the rest span. PMID- 15545299 TI - Genome-wide analysis of the unfolded protein response in fibroblasts from congenital disorders of glycosylation type-I patients. AB - Congenital disorders of glycosylation (CDG) are a family of diseases characterized by defects of N-linked glycosylation. In CDG-I, several genetic defects cause a shortage of dolichol-linked oligosaccharides, which leads to underglycosylation of nascent glycoproteins. N-linked glycosylation is important for proper folding and trafficking of glycoproteins. Inhibition of glycosylation results in the buildup of misfolded proteins in the endoplasmic reticulum, which induces a protective reaction known as the unfolded protein response (UPR). To investigate whether UPR components are induced in CDG, we have performed a transcriptome analysis of primary fibroblasts from unaffected control subjects and from CDG-I patients using oligonucleotide gene expression arrays. The stress imposed by CDG was also compared with the stress induced by tunicamycin and glucose deprivation. Whereas tunicamycin elicited a strong transcriptional response typical for the UPR, CDG fibroblasts displayed a qualitatively similar yet moderate induction of genes encoding components of the UPR. Among these genes, the PERK kinase inhibitor DNAJC3/P58(IPK) gene showed the highest induction throughout all CDG-I types tested. This was paralleled by elevated expression of genes involved in amino acid biosynthesis and transport, which defined a new component of the cellular response to glycosylation stress. PMID- 15545300 TI - TNF receptor 1 is involved in the induction of apoptosis by the cyclin dependent kinase inhibitor p27Kip1 in the prostate cancer cell line PC-3. AB - Loss of p27Kip1, a cyclin-dependent kinase inhibitor, is observed in aggressive prostate cancers. We demonstrated that intratumoral injections of recombinant adenovirus overexpressing p27Kip1 (Adp27) reduced the growth of prostate cancer xenografts in nude mice. Presently, we studied the mechanism(s) of cell death induced by Adp27 in prostate cancer cell line PC-3. Cells were infected with Adp27 and compared with those infected by empty virus or were non-infected. Cell cycle and typical markers of apoptosis were analyzed by flow cytometry in the presence of the following reagents: cycloheximide, pan-caspase inhibitor ZVAD fmk, neutralizing anti-TNFR1, and anti-TNFR2. Overexpression of p27Kip1 protein and cell cycle arrest were noted within 24 h after Adp27-infection. Sub-G1 fraction, chromatin margination, and phosphatidylserine exposure were evident by the third day of treatment. Cycloheximide elevated sub-G1 fraction in Adp27 infected cells by threefold, while ZVAD-fmk reduced sub-G1 to control levels. Caspase-dependent apoptosis occurred in a third of the population, while two thirds were ZVAD-fmk insensitive but TUNEL-positive. Flow cytometry showed increased expression of TNFR1 and TNFR2 in Adp27-infected cells. Neutralizing anti-TNFR1 decreased TUNEL-positive score, while anti-TNFR2 did not affect p27Kip1-induced apoptosis. This is the first report showing that p27Kip1 induces caspase-dependent and -independent stages of cell death that may involve TNF signaling through TNFR1. PMID- 15545301 TI - The modulation of caveolin-1 expression controls satellite cell activation during muscle repair. AB - We have previously shown that caveolin-1, the principal structural protein component of caveolar membrane domains, inhibits cellular proliferation and induces cell cycle arrest. We demonstrate here for the first time that caveolin-1 is expressed in satellite cells but not in mature muscle fibers. Satellite cells are quiescent myogenic precursors that, after muscle injury, become mitotically active, proliferate, and fuse together or, to existing myofibers, to form new muscle fibers. We show that down-regulation of caveolin-1 expression occurs in satellite cells/myogenic precursor cells (MPCs) during muscle regeneration and that hepatocyte growth factor, which is produced after muscle injury, down regulates caveolin-1. We also demonstrate that down-regulation of endogenous caveolin-1 expression activates ERK and that activation of the p42/44 MAP kinase pathway is necessary to promote muscle regeneration. Finally, we show that overexpression of caveolin-1 inhibits muscle repair mechanisms both in vitro and in vivo. Taken together, these results propose caveolin-1 as a novel regulator of satellite cell functions and suggest that the following signaling pathway modulates satellite cell activation during muscle repair: injured fibers release HGF --> HGF down-regulates caveolin-1 protein expression --> down-regulation of caveolin-1 activates ERK --> activation of ERK promotes muscle repair by stimulating the proliferation and migration of MPCs toward the wounded area. PMID- 15545302 TI - Lycopene reduced gene expression of steroid targets and inflammatory markers in normal rat prostate. AB - Epidemiological evidence links consumption of lycopene, the red carotenoid of tomato, to reduced prostate cancer risk. We investigated the effect of lycopene in normal prostate tissue to gain insight into the mechanisms, by which lycopene can contribute to primary prostate cancer prevention. We supplemented young rats with 200 ppm lycopene for up to 8 wk, measured the uptake into individual prostate lobes, and analyzed lycopene-induced gene regulations in dorsal and lateral lobes after 8 wk of supplementation. Lycopene accumulated in all four prostate lobes over time, with all-trans lycopene being the predominant isoform. The lateral lobe showed a significantly higher total lycopene content than the other prostate lobes. Transcriptomics analysis revealed that lycopene treatment mildly but significantly reduced gene expression of androgen-metabolizing enzymes and androgen targets. Moreover, local expression of IGF-I was decreased in the lateral lobe. Lycopene also consistently reduced transcript levels of proinflammatory cytokines, immunoglobulins, and immunoglobulin receptors in the lateral lobe. This indicates that lycopene reduced inflammatory signals in the lateral prostate lobe. In summary, we show for the first time that lycopene reduced local prostatic androgen signaling, IGF-I expression, and basal inflammatory signals in normal prostate tissue. All of these mechanisms can contribute to the epidemiologically observed prostate cancer risk reduction by lycopene. PMID- 15545303 TI - Directed differentiation of embryonic stem cells into dorsal interneurons. AB - During neural development caudalization and dorsoventral patterning of the neural tube is directed by several inductive factors including retinoic acid, sonic hedgehog (Shh), bone morphogenetic proteins (BMPs), and Wnt signaling. The purpose of the current study was to investigate whether dorsal interneurons specific for the spinal cord can be generated from mouse embryonic stem (ES) cells using known inductive signals. Here we show that specific combination of developmental signaling molecules including all trans-retinoic acid, Shh, bone morphogenetic protein 2 (BMP2), and Wnt3A can direct differentiation of ES cells into dorsal interneurons possessing appropriate neuronal markers, synaptic proteins and functional neurotransmitter machineries. We introduce a concept that Wnt3A morphogenic action relies on crosstalk with both Shh and BMP2 signaling pathways. PMID- 15545304 TI - Dopamine-1 receptor agonist: renal effects and its potential role in the management of radiocontrast-induced nephropathy. AB - Radiocontrast-induced nephropathy remains the third leading cause of hospital acquired acute renal failure. Once established, this syndrome is associated with increased morbidity and mortality as well as increased health care costs. Recently, studies have been initiated to evaluate the potential of a selective dopamine-1 receptor agonist (fenoldopam) in ameliorating radiocontrast-induced renal failure. Selective dopamine-1 receptor agonists exhibit many desirable renal effects that support their use for the prophylaxis of radiocontrast-induced nephropathy, including decreases in renal vascular resistance and increases in renal blood flow, glomerular filtration, and sodium and water excretion. Several reports have documented a beneficial effect of fenoldopam administration in attenuating radiocontrast-induced nephropathy. In contrast, a recent multicenter, randomized study did not demonstrate a renoprotective effect of fenoldopam against radiocontrast-induced nephropathy. The presence of multiple confounders, however, precludes a definitive conclusion regarding the ability of fenoldopam to protect against radiocontrast-induced nephropathy. Additional studies are needed to properly evaluate the role of fenoldopam in radiocontrast-induced nephropathy prophylaxis. PMID- 15545305 TI - Utility of sparse concentration sampling for citalopram in elderly clinical trial subjects. AB - The objective of this study was to evaluate whether the disposition of the selective serotonin reuptake inhibitor, citalopram, could be robustly captured using 1 to 2 concentration samples per subject in 106 patients participating in 2 clinical trials. Nonlinear mixed-effects modeling was used to evaluate the pharmacokinetic parameters describing citalopram's disposition. Both a prior established 2-compartment model and a de novo 1-compartment pharmacokinetic model were used. Covariates assessed were concomitant medications, race, sex, age (22 93 years), and weight. Covariates affecting disposition were assessed separately and then combined in a stepwise manner. Pharmacokinetic characteristics of citalopram were well captured using this sparse sampling design. Two covariates (age and weight) had a significant effect on the clearance and volume of distribution in both the 1- and 2-compartment pharmacokinetic models. Clearance decreased 0.23 L/h for every year of age and increased 0.14 L/h per kilogram body weight. It was concluded that hyper-sparse sampling designs are adequate to support population pharmacokinetic analysis in clinically treated populations. This is particularly valuable for populations such as the elderly, who are not typically available for pharmacokinetic studies. PMID- 15545306 TI - DOV 216,303, a "triple" reuptake inhibitor: safety, tolerability, and pharmacokinetic profile. AB - This report describes the first evaluation in humans of DOV 216,303, a putative antidepressive that inhibits the reuptake of norepinephrine, serotonin, and dopamine. Subjects received single oral doses of 5 to 150 mg of DOV 216,303 or placebo. At 150 mg, 4 of 7 subjects reported gastrointestinal disturbances. In the multiple-dose phase of the evaluation, subjects received total daily doses of 50, 75, or 100 mg of DOV 216,303 or placebo for 10 days. At a total daily dose of 100 mg, gastrointestinal disturbances were reported in 4 of 6 volunteers. In both the single- and multiple-dose evaluations, no significant changes were noted in vital signs, electrocardiogram, hematology, or clinical chemistry. DOV 216,303 was rapidly absorbed (plasma t(max) of 0.7-1.2 hours and t(1/2) of 3.3-4.4 hours), with dose-proportional C(max) and AUC values. Furthermore, no remarkable difference was apparent in either the C(max) or AUC(tau) of DOV 216,303 following 1 and 10 days of dosing. The present results demonstrate that DOV 216,303 is safe and well tolerated both at single doses of up to 100 mg and multiple doses of up to 100 mg/day for 10 days. Plasma concentrations of DOV 216,303 after doses > 10 mg exceed its reported IC(50) values for inhibition of biogenic amine reuptake. PMID- 15545307 TI - Pharmacokinetics, safety, and tolerability of R411, a dual alpha4beta1 alpha4beta7 integrin antagonist after oral administration at single and multiple once-daily ascending doses in healthy volunteers. AB - R411 is a dual alpha4beta1-alpha4beta7 integrin antagonist under development for the treatment of chronic asthma. The objective of this study was to investigate the pharmacokinetics and safety of R411 and its active metabolite, RO0270608, in humans. A 3-part phase I trial was conducted in 132 healthy volunteers: (1) 12 subjects received 200 mg R411 as a single oral dose or 100 mg RO0270608 as an intravenous infusion in a 1-sequence crossover design; (2) 7 groups of 10 subjects received 1 of 7 single oral doses of R411 (10-1200 mg) in a parallel, placebo-controlled, ascending adaptive dose design; and (3) 5 groups of 10 subjects each received repeated oral qd doses of R411 (50-900 mg) for up to 3 weeks in a parallel, placebo-controlled, ascending adaptive dose design. The absolute bioavailability of RO0270608 (mean +/- standard deviation) after oral administration of R411 was 27% +/- 4%, and the terminal half-life was 7.33 +/- 2.29 hours. After IV infusion of RO0270608, total clearance (mean +/- standard deviation) was 19.4 +/- 7.1 L/h, and the volume of distribution was 93.1 +/- 36.1 L. After single ascending oral doses of R411, area under the concentration-time curve from 0 to infinity of active metabolite RO0270608 increased proportionally from 150 to 1200 mg (P > .05). Following repeated administration, the oral clearance was independent of time. No drug accumulation was observed, and no safety concerns were revealed up to a dose of 900 mg after up to 3 weeks of treatment. PMID- 15545308 TI - The effect of food on the pharmacokinetics of a dual angiotensin-converting enzyme/neutral endopeptidase inhibitor, M100240. AB - M100240 is a thioester of MDL 100,173, a dual angiotensin-converting enzyme (ACE)/neutral endopeptidase (NEP) inhibitor. Clinical studies have shown that M100240 is capable of decreasing ACE activity and angiotensin II concentrations while increasing plasma renin activity and potentiating the effects of atrial natriuretic peptide. This may result in a unique treatment benefit in disease states characterized by intravascular volume or sodium overload or increased venous pressure. The pharmacokinetics of MDL 100,173 were evaluated in 30 healthy subjects in an open-label, randomized, 2-period crossover design. Subjects received a single oral dose of 50 mg of M100240 administered with a high-fat meal and separately under fasted conditions. Serial plasma concentrations of M100240 and MDL 100,173 were analyzed, and pharmacokinetic parameters were calculated with noncompartmental methods. The intrasubject percent coefficient of variation for MDL 100,173 C(max) and AUC(0-24h) were less than 20%, indicating that this agent is a moderately variable drug. Although AUC(0-24h) was within the protocol defined range of 80% to 125%, the lower limit of the 90% confidence interval for C(max) fell outside of the 70% to 143% range. Absence of a food effect on the pharmacokinetic profile of 50 mg of M100240 could therefore not be demonstrated. This finding is not surprising based on the documented food effect with the sulfhydryl-containing ACE inhibitor, captopril. Clinical significance of this pharmacokinetic food effect is unlikely, as the magnitude of pharmacodynamic response is probably better correlated with AUC than a single-point determination of C(max). Single oral doses of 50 mg of M100240 were safe and well tolerated under fed and fasted conditions. PMID- 15545309 TI - Dose-dependent alternations in the pharmacokinetics of olanzapine during coadministration of fluvoxamine in patients with schizophrenia. AB - Olanzapine, an atypical antipsychotic agent, is a substrate of the cytochrome P4501A2 (CYP1A2) enzyme. Administration of a potent CYP1A2 inhibitor (eg, fluvoxamine) may alter the pharmacokinetics of olanzapine. This study investigated the pharmacokinetic interactions between olanzapine and fluvoxamine in patients with schizophrenia. Ten male smokers were administrated a single dose of olanzapine 10 mg at baseline, followed by 2 weeks of fluvoxamine 50 mg/day and another 2 weeks of fluvoxamine 100 mg/day. Olanzapine 10 mg was given at day 10 during each fluvoxamine treatment. After pretreatment with fluvoxamine, the area under the curve, maximal plasma concentration, and half-time of olanzapine were significantly increased by 30% to 55%, 12% to 64%, and 25% to 32%, respectively. Volume of distribution and apparent clearance were significantly reduced by 4% to 26% and 26% t O 38%, respectively, after administration of fluvoxamine. Increases in area under the plasma concentration-time curve from time 0 to infinity appear to be dose dependent. Presumably, altered olanzapine pharmacokinetics are attributed to the inhibition of CYP1A2. Patients treated with the combination of olanzapine and fluvoxamine should be monitored carefully. PMID- 15545310 TI - A randomized crossover study investigating the influence of ranitidine or omeprazole on the pharmacokinetics of cephalexin monohydrate. AB - Limited data characterize pharmacokinetic interactions between cephalexin and ranitidine, and no data exist for an interaction with proton pump inhibitors. The purpose of this study was to investigate the effects of ranitidine or omeprazole administration on the pharmacokinetics and pharmacodynamics of cephalexin. A randomized single- and multiple-dose crossover study was conducted in healthy subjects ingesting cephalexin before and after steady-state administration of ranitidine or omeprazole. Time-concentration profiles were determined and pharmacokinetic parameters were characterized using noncompartmental methods. Pharmacokinetic data were analyzed in accordance with the two 1-sided test for bioequivalence. The percentage of time that serum concentrations remain above the MIC(90) during the dosing interval (T > MIC(90)) for Streptococcus pyogenes and Staphylococcus aureus associated with the pharmacokinetic profiles was calculated. The coadministration of cephalexin with ranitidine or omeprazole resulted in relatively minor changes in C(max), AUC(infinity), t(1/2), or CL/F. t(max) was significantly prolonged when cephalexin was administered with ranitidine or omeprazole. Suboptimal T > MIC(90) was observed for cephalexin irrespective of acid suppression. Delay in absorption of cephalexin resulted in a decrease in the percentage of T > MIC(90) for certain acid-suppressive regimens and pathogen combinations. With the exception of an increase in t(max), there were no significant pharmacokinetic interactions between cephalexin and ranitidine or omeprazole. Delayed t(max) associated with acid suppression may result in a diminished T > MIC(90). PMID- 15545311 TI - Assessment of in vivo CYP2D6 activity: differential sensitivity of commonly used probes to urine pH. AB - Drug/metabolite ratios (MRs) are used as in vivo markers of enzyme activity. The ratios are potentially confounded by the renal clearance of the drug (urine-based MRs) or metabolite (plasma-based MRs). The authors have investigated the relative sensitivity of urinary MR of 3 in vivo probe substrates of CYP2D6 debrisoquine (DB), dextromethorphan (DM), and metoprolol (MP) to changes in urine pH. Three groups of healthy volunteers each comprising 12 individuals were given DB (10 mg), DM (25 mg), or MP (100 mg) on 3 occasions. In 1 study arm, urine was acidified by the oral intake of ammonium chloride; in another, it was alkalinized by intake of sodium bicarbonate; and in the third, urine pH was uncontrolled. Urinary MP/alpha-hydroxy-MP, DM/dextrorphan, and DB/4-hydroxy-DB ratios were calculated. The mean(geo) MR for DB was not significantly different in any of the study arms, whereas those for MP and DM were significantly different under acidified and alkalinized urine conditions compared to uncontrolled urine pH (P < .01) and were correlated with urine pH (P < .001). Without control of urine pH, in vivo estimates of CYP2D6 metabolic activity are likely to be less precise using DM or MP as probe substrates compared to DB. Although this is unlikely to cause any problem in distinguishing the large functional differences in CYP2D6 in poor metabolizer (PM) and extensive metabolizer (EM) phenotypes, this may contribute to difficulties in differentiating in vivo metabolic activity among allelic variants within the overall CYP2D6 EM phenotype using MP or DM. However, because DB is not available in many countries (eg, United States), alternative in vivo markers of CYP2D6 with low sensitivity to urine pH should be sought. PMID- 15545312 TI - Metabolism by N-acetyltransferase 1 in vitro and in healthy volunteers: a prototype for targeted inhibition. AB - Inhibition of drug metabolism is generally avoided but can be useful in limited circumstances, such as reducing the formation of toxic metabolites. Acetylation is a major pathway for drug elimination that can also convert substrates into toxic species, including carcinogens. Sulfamethoxazole, a widely used antibiotic, is metabolized via arylamine N-acetyltransferase 1. p-Aminosalicylate, used for antitubercular treatment, is also metabolized by N-acetyltransferase 1 and could potentially inhibit sulfamethoxazole metabolism. Human hepatocytes from 4 donors were incubated in vitro with sulfamethoxazole and paminosalicylate at clinically achievable concentrations. p-Aminosalicylate competitively reduced the acetylation of sulfamethoxazole in vitro by 61% to 83% at 200 microM. Four healthy volunteers were studied following doses of 500 mg sulfamethoxazole either alone or during administration of paminosalicylate (4 g ter in die). Plasma concentrations of paminosalicylate exceeded 100 microM. With each subject as his or her own control, p-aminosalicylate reduced by 5-fold the ratio of plasma concentrations of acetylsulfamethoxazole relative to parent drug (P < .001). Metabolic drug-drug interaction studies in vitro successfully predicted inhibition of acetylation via N-acetyltransferase 1 in vivo. Although no specific toxic species was investigated in this work, the potential was demonstrated for improving the therapeutic index of drugs that have toxic metabolites. PMID- 15545313 TI - Interindividual and intraindividual variability of the urinary 6beta Hydroxycortisol/Cortisol ratio in Chinese subjects: implications of its use for evaluating CYP3A activity. AB - The present study determined the interindividual and intrandividual variability of the urinary 6beta-hydroxycortisol/cortisol ratio, a useful marker for CYP3A induction and inhibition in Chinese subjects. The study consisted of 2 parts. In part I, 82 healthy male Chinese subjects underwent 3 study sessions, each separated by a 1-week interval. In part II, 20 subjects who initially completed part I underwent another 3 sessions over a period of 3 to 4 months. During each session, a first-morning urine specimen was collected from each subject for the quantification of urinary concentrations of cortisol and 6beta-hydroxycortisol. There were no significant differences in the mean 6beta-hydroxycortisol/cortisol ratios among the 3 sessions (P > .05, 1-way analysis of variance) for both part I and part II of the study. A normal distribution of the 6beta hydroxycortisol/cortisol ratio was observed (P = .849, Kolmogorov-Smirnov test). This ratio varied 30-fold (range, 0.76-23.23) among the study subjects. The mean intraindividual variabilities during the short (3-week) and long (3- to 4-month) periods were 30.9% +/- 17.5% and 32.2% +/- 17.1%, respectively. The genetic fraction contributing to the observed variability in the 6beta hydroxycortisol/cortisol ratio was estimated to be 0.91. The genetic component is likely to contribute significantly to the variability of the 6beta hydroxycortisol/cortisol ratio, and such variability should be considered when the ratio is used to evaluate CYP3A induction or inhibition in a given ethnic population. PMID- 15545314 TI - Analgesic effect of sustained-release flurbiprofen administered at the site of tissue injury in the oral surgery model. AB - Nonsteroidal anti-inflammatory drugs produce their analgesic and adverse effects through interaction with cyclooxygenase in a variety of tissues. The authors evaluated the therapeutic potential of administering a sustained-release formulation of flurbiprofen into a surgical wound following oral surgery to produce analgesia at the site of injury while minimizing exposure to potential targets for toxicity. Subjects (N = 98) received 1 of 8 treatments: flurbiprofen in a microparticle formulation in doses of 3.125 mg, 6.25 mg, 12.5 mg, 25 mg, or 50 mg; PO flurbiprofen 25 mg or 50 mg; or placebo. The flurbiprofen microparticle formulation or matching placebo was placed into the extraction sites at the end of surgery (removal of 2 lower impacted third molars). The sum of the pain visual analog scale over the 6-hour observation period demonstrated significantly less pain (P < .05) for flurbiprofen microparticle in comparison with placebo. Fewer subjects remedicated in the flurbiprofen microparticle drug groups, primarily for the 12.5-mg and higher doses. The incidence of adverse effects and local complications did not differ across groups. These data suggest that direct administration of flurbiprofen in a microparticle formulation at a site of tissue injury delays the onset and lowers the intensity of postoperative pain at lower doses than usually administered orally. PMID- 15545315 TI - Effect of modafinil on acute pain: a randomized double-blind crossover study. PMID- 15545316 TI - MT1-MMP-dependent neovessel formation within the confines of the three dimensional extracellular matrix. AB - During angiogenesis, endothelial cells initiate a tissue-invasive program within an interstitial matrix comprised largely of type I collagen. Extracellular matrix degradative enzymes, including the matrix metalloproteinases (MMPs) MMP-2 and MMP 9, are thought to play key roles in angiogenesis by binding to docking sites on the cell surface after activation by plasmin- and/or membrane-type (MT) 1-MMP dependent processes. To identify proteinases critical to neovessel formation, an ex vivo model of angiogenesis has been established wherein tissue explants from gene-targeted mice are embedded within a three-dimensional, type I collagen matrix. Unexpectedly, neither MMP-2, MMP-9, their cognate cell-surface receptors (i.e., beta3 integrin and CD44), nor plasminogen are essential for collagenolytic activity, endothelial cell invasion, or neovessel formation. Instead, the membrane-anchored MMP, MT1-MMP, confers endothelial cells with the ability to express invasive and tubulogenic activity in a collagen-rich milieu, in vitro or in vivo, where it plays an indispensable role in driving neovessel formation. PMID- 15545317 TI - Selective apoptosis of pluripotent mouse and human stem cells by novel ceramide analogues prevents teratoma formation and enriches for neural precursors in ES cell-derived neural transplants. AB - The formation of stem cell-derived tumors (teratomas) is observed when engrafting undifferentiated embryonic stem (ES) cells, embryoid body-derived cells (EBCs), or mammalian embryos and is a significant obstacle to stem cell therapy. We show that in tumors formed after engraftment of EBCs into mouse brain, expression of the pluripotency marker Oct-4 colocalized with that of prostate apoptosis response-4 (PAR-4), a protein mediating ceramide-induced apoptosis during neural differentiation of ES cells. We tested the ability of the novel ceramide analogue N-oleoyl serinol (S18) to eliminate mouse and human Oct-4(+)/PAR-4(+) cells and to increase the proportion of nestin(+) neuroprogenitors in EBC-derived cell cultures and grafts. S18-treated EBCs persisted in the hippocampal area and showed neuronal lineage differentiation as indicated by the expression of beta tubulin III. However, untreated cells formed numerous teratomas that contained derivatives of endoderm, mesoderm, and ectoderm. Our results show for the first time that ceramide-induced apoptosis eliminates residual, pluripotent EBCs, prevents teratoma formation, and enriches the EBCs for cells that undergo neural differentiation after transplantation. PMID- 15545318 TI - Ubiquitin charging of human class III ubiquitin-conjugating enzymes triggers their nuclear import. AB - Ubiquitin is a small polypeptide that is conjugated to proteins and commonly serves as a degradation signal. The attachment of ubiquitin (Ub) to a substrate proceeds through a multi-enzyme cascade involving an activating enzyme (E1), a conjugating enzyme (E2), and a protein ligase (E3). We previously demonstrated that a murine E2, UbcM2, is imported into nuclei by the transport receptor importin-11. We now show that the import mechanism for UbcM2 and two other human class III E2s (UbcH6 and UBE2E2) uniquely requires the covalent attachment of Ub to the active site cysteine of these enzymes. This coupling of E2 activation and transport arises from the selective interaction of importin-11 with the Ub-loaded forms of these enzymes. Together, these findings reveal that Ub charging can function as a nuclear import trigger, and identify a novel link between E2 regulation and karyopherin-mediated transport. PMID- 15545319 TI - Control of mitochondrial motility and distribution by the calcium signal: a homeostatic circuit. AB - Mitochondria are dynamic organelles in cells. The control of mitochondrial motility by signaling mechanisms and the significance of rapid changes in motility remains elusive. In cardiac myoblasts, mitochondria were observed close to the microtubular array and displayed both short- and long-range movements along microtubules. By clamping cytoplasmic [Ca2+] ([Ca2+]c) at various levels, mitochondrial motility was found to be regulated by Ca2+ in the physiological range. Maximal movement was obtained at resting [Ca2+]c with complete arrest at 1 2 microM. Movement was fully recovered by returning to resting [Ca2+]c, and inhibition could be repeated with no apparent desensitization. The inositol 1,4,5 trisphosphate- or ryanodine receptor-mediated [Ca2+]c signal also induced a decrease in mitochondrial motility. This decrease followed the spatial and temporal pattern of the [Ca2+]c signal. Diminished mitochondrial motility in the region of the [Ca2+]c rise promotes recruitment of mitochondria to enhance local Ca2+ buffering and energy supply. This mechanism may provide a novel homeostatic circuit in calcium signaling. PMID- 15545321 TI - Pex7p translocates in and out of peroxisomes in Saccharomyces cerevisiae. AB - Pex7p is the soluble receptor responsible for importing into peroxisomes newly synthesized proteins bearing a type 2 peroxisomal targeting sequence. We observe that appending GFP to Pex7p's COOH terminus shifts Pex7p's intracellular distribution from predominantly cytosolic to predominantly peroxisomal in Saccharomyces cerevisiae. Cleavage of the link between Pex7p and GFP within peroxisomes liberates GFP, which remains inside the organelle, and Pex7p, which exits to the cytosol. The reexported Pex7p is functional, resulting in import of thiolase into peroxisomes and improved growth of the yeast on oleic acid. These results support the "extended shuttle" model of peroxisome import receptor function and open the way to future studies of receptor export. PMID- 15545320 TI - The RNA-binding protein SUP-12 controls muscle-specific splicing of the ADF/cofilin pre-mRNA in C. elegans. AB - Tissue-specific alternative pre-mRNA splicing is essential for increasing diversity of functionally different gene products. In Caenorhabditis elegans, UNC 60A and UNC-60B, nonmuscle and muscle isoforms of actin depolymerizing factor (ADF)/cofilin, are expressed by alternative splicing of unc-60 and regulate distinct actin-dependent developmental processes. We report that SUP-12, a member of a new family of RNA recognition motif (RRM) proteins, including SEB-4, regulates muscle-specific splicing of unc-60. In sup-12 mutants, expression of UNC-60B is decreased, whereas UNC-60A is up-regulated in muscle. sup-12 mutations strongly suppress muscle defects in unc-60B mutants by allowing expression of UNC 60A in muscle that can substitute for UNC-60B, thus unmasking their functional redundancy. SUP-12 is expressed in muscle and localized to the nuclei in a speckled pattern. The RRM domain of SUP-12 binds to several sites of the unc-60 pre-mRNA including the UG repeats near the 3'-splice site in the first intron. Our results suggest that SUP-12 is a novel tissue-specific splicing factor and regulates functional redundancy among ADF/cofilin isoforms. PMID- 15545322 TI - Impact of telomerase ablation on organismal viability, aging, and tumorigenesis in mice lacking the DNA repair proteins PARP-1, Ku86, or DNA-PKcs. AB - The DNA repair proteins poly(ADP-ribose) polymerase-1 (PARP-1), Ku86, and catalytic subunit of DNA-PK (DNA-PKcs) have been involved in telomere metabolism. To genetically dissect the impact of these activities on telomere function, as well as organismal cancer and aging, we have generated mice doubly deficient for both telomerase and any of the mentioned DNA repair proteins, PARP-1, Ku86, or DNA-PKcs. First, we show that abrogation of PARP-1 in the absence of telomerase does not affect the rate of telomere shortening, telomere capping, or organismal viability compared with single telomerase-deficient controls. Thus, PARP-1 does not have a major role in telomere metabolism, not even in the context of telomerase deficiency. In contrast, mice doubly deficient for telomerase and either Ku86 or DNA-PKcs manifest accelerated loss of organismal viability compared with single telomerase-deficient mice. Interestingly, this loss of organismal viability correlates with proliferative defects and age-related pathologies, but not with increased incidence of cancer. These results support the notion that absence of telomerase and short telomeres in combination with DNA repair deficiencies accelerate the aging process without impacting on tumorigenesis. PMID- 15545323 TI - Neuroimaging in geriatric psychiatry. PMID- 15545324 TI - Systematic review of the diagnostic accuracy of 99mTc-HMPAO-SPECT in dementia. AB - OBJECTIVE: The authors sought to determine the diagnostic accuracy of 99mTc-HMPAO SPECT in discriminating between Alzheimer disease (AD) and other dementias. METHODS: Articles published between 1985 and 2002 were retrieved systematically from MEDLINE and EMBASE, cross-referencing with personal collections and 13 narrative reviews. Of 301 studies identified, 48 survived exclusion criteria and contained extractable data. Two authors independently assessed and graded the methodology of all included studies. Diagnostic comparison groups included vascular dementia (VD; 13 studies), fronto-temporal dementia (FTD; 7 studies), normal healthy volunteers (27 studies), and non-dementia patients (13 studies). Where statistically justified, groups were pooled in a metaanalysis; summary receiver operating curves were constructed; and heterogeneity across studies examined by regression of the diagnostic odds ratio. RESULTS: The pooled weighted sensitivity of 99mTc-HMPAO-SPECT in discriminating clinically defined AD from VD was 71.3%; its specificity was 75.9%. The pooled weighted sensitivity and specificity for AD versus FTD were 71.5% and 78.2%, respectively. Variation in outcome across studies was not found to be attributable to any single factor. CONCLUSION: Pathological verification studies suggest that clinical criteria may be more sensitive in detecting AD than brain SPECT (81% versus 74%). However, SPECT studies provide a higher specificity against other types of dementia than clinical criteria (91% versus 70%). SPECT may, therefore, be helpful in the differential diagnosis of AD. Clinical follow-up studies are urgently required to establish its predictive validity with regard to natural history and treatment response. PMID- 15545325 TI - Late-life alcoholism: issues relevant to the geriatric psychiatrist. AB - Alcohol use and abuse are often overlooked as an important clinical issue in older adults. However, with a rapidly growing population of older adults who grew up during an era of increased illicit drug and alcohol use, there are reasons to believe that there will be a greater impact of alcohol abuse or dependence on the next generation of older adults. Because older adults seek the majority of behavioral health care either in primary care settings or from mental health providers, it is necessary for primary care providers and geriatric mental health providers to become proficient at assessing and providing interventions. This article discusses the state of research in this area and reviews clinically relevant concepts related to identifying, assessing, and treating older adults with alcohol-related disability. PMID- 15545326 TI - In-vivo imaging of Alzheimer disease beta-amyloid with [11C]SB-13 PET. AB - OBJECTIVE: In-vivo imaging of beta-amyloid plaques (Abeta) may improve both early detection of Alzheimer disease (AD) and efficacy assessment of new treatments for AD. The authors' aim was to develop a novel Abeta-specific positron-emission tomography (PET) tracer. METHODS: Five female AD patients (54-77 years old) and six healthy female comparison subjects (53-74 years old), completed 2-hour PET scans after intravenous injection of 10 mCi of both the stilbene [11C]SB-13 and the benzothiazole [11C]6-OH-BTA-1 (also known as [11C]PIB). Kinetic analyses were performed on the resulting time-activity curves to derive Abeta binding-potential estimates, using as input function either the unmetabolized tracer concentration in venous plasma from a two-tissue compartment model or the density of radioactivity in the cerebellum. Authors compared the binding characteristics of the two radiotracers. RESULTS: The two radiotracers demonstrated similar binding properties with respect to regional distribution of retention (increased retention in the frontal and posterior temporal-inferior parietal association cortices in the AD patients, but not in the comparison subjects). Our preliminary PET data indicate that [11C]SB-13 may be similar to [11C]PIB in discriminating AD patients from comparison subjects. CONCLUSIONS: [11C]SB-13 is an effective PET tracer for fibrillar Abeta imaging in vivo, with similar performance as [11C]PIB. Future research directions include evaluation of tracer in larger AD patient samples and in subjects with amnestic mild cognitive impairment, evaluation of arterial input function, and comparison with other tracers, such as [18F]FDG as they relate to cognitive functioning. PMID- 15545327 TI - Altered PET functional brain responses in cognitively intact elderly persons at risk for Alzheimer disease (carriers of the epsilon4 allele). AB - OBJECTIVE: Few previous studies have investigated the association between APOE genotype and brain activation during performance of cognitive tasks in healthy middle-aged and elderly subjects, and the results have been mixed. The authors investigated APOE-mediated differential brain activation in a group of healthy elderly subjects. METHODS: Using H215O positron emission tomography (PET), they imaged 32 healthy subjects (26 non-epsilon4 carriers and 6 epsilon4 carriers) performing a serial shape-recognition memory task under two conditions: Simple Demand (SD), in which one shape was presented in each study trial, and Titrated Demand (TD), in which study list length was adjusted so that each subject recognized words at approximately 75% accuracy. Multiple-regression analyses were performed, with the "activation" difference (TD-SD PET counts) as the dependent variable and the APOE genotype (presence versus absence of the epsilon4 allele) as the independent variable. RESULTS: Compared with non-carriers, epsilon4 carriers exhibited significantly decreased TD-SD activation differences in the left superior temporal, right superior frontal, left postcental, left precuneus, and posterior cingulate gyrus because epsilon4 carriers (versus non-carriers) showed increased activation during the SD and decreased activation during the TD condition. CONCLUSION: Patterns of brain activation during a nonverbal memory task differed as a function of APOE genotype and, therefore, of genetic risk for Alzheimer disease (AD). Differences in activation were not a reflection of task difficulty, but indicate memory-related altered cognitive processing. Brain regions with decreased activation in the epsilon4 subjects may result from subclinical incipient AD pathology and/or APOE-related neurophysiologic heterogeneity. PMID- 15545328 TI - A volumetric study of MRI signal hyperintensities in late-life depression. AB - OBJECTIVE: An increase in white-matter lesions has been previously described in older subjects with depression. The authors investigated whether the regional location varied between depressed and normal subjects and determined the relationship of magnetic resonance (MR) signal hyperintensities to known clinical risk factors for vascular disease. METHODS: Authors used automated image processing software to determine volumes of signal hyperintensities from MR brain scans of older people with depression (N=29; mean age: 76 years) and normal subjects of similar age (N=32). RESULTS: Overall, subjects with depression had a significantly greater frontal-lobe white-matter lesion volume than normal subjects (0.35% versus 0.22%). However, after excluding subjects with hypertension, diabetes, or ischemic heart disease (leaving 14 depressed and 15 normal subjects), we found even greater differences between groups, with a larger volume of MR signal hyperintensities in the frontal region of the depressed group, but no difference in the basal ganglia or parietal and occipital lobes. CONCLUSION: The results support the "vascular depression" hypothesis and suggest that those with depression but without traditional vascular risk factors may be much more susceptible to cerebrovascular disease than normal subjects. The mechanisms for this increased susceptibility remain to be determined. PMID- 15545329 TI - Hippocampal volume measurement in older adults with bipolar disorder. AB - OBJECTIVE: Decreased hippocampal volumes have been noted in unipolar depressed subjects, especially in elderly patients and those with cognitive impairment. Initial studies of mixed-aged bipolar subjects and controls have had conflicting findings, with most noting no difference; however this region has not been examined in older bipolar subjects. METHODS: The authors examined the hippocampal volumes of 36 older bipolar subjects (mean age: 58 years) and 29 older normal comparison (NC) subjects (mean age: 61), using logistic-regression analyses to control for age and gender. Differences between late- and early-onset (before age 45) bipolar subjects were also examined. RESULTS: The left hippocampus was noted to be enlarged in older bipolar subjects, compared with the older NC group (sex and age controlled). No differences were noted in hippocampal volumes by age at onset nor number of previous episodes. The increase in hippocampal volume may be associated with the use of lithium, but not valproic acid. CONCLUSIONS: Left hippocampal volume is increased in older bipolar subjects compared with NC subjects. The differences were not explained by age at onset, current mood state, or cognitive status, but may be associated with exposure to lithium. This finding would support previous observations about the neural-plasticity effect of lithium. PMID- 15545330 TI - Pilot study on the effect of estrogen replacement therapy on brain dopamine transporter availability in healthy, postmenopausal women. AB - OBJECTIVE: Authors investigated the association between estrogen replacement therapy (ERT) and dopamine transporter (DAT) availability in women. METHODS: Thirteen postmenopausal women were administered ERT and underwent neuroimaging, using single-photon emission computed tomography (SPECT) and [99mTc]TRODAT-1, a radioligand that binds DAT. In this 6-week pilot study, subjects underwent SPECT before ERT, after 4 weeks of 0.625 mg/day of conjugated estrogens (CEE), and after an additional 2 weeks of 0.625 mg/day CEE plus 10 mg/day of medroxyprogesterone acetate. Specific uptake values (SUVs) of [99mTc]TRODAT-1 were calculated for the caudate and putamen. RESULTS: When compared with baseline values, [99mTc]TRODAT-1 binding demonstrated a modest, but statistically significant, increase in the left anterior putamen after 4 weeks of CEE. After the 6-week ERT intervention, both the left and right anterior putamen demonstrated an increase in SUVs. CONCLUSION: Short-term administration of ERT in postmenopausal women is associated with a modest increase in DAT in the putamen. These findings may further the understanding of how ERT is associated with improvement in Parkinson's disease and late-onset schizophrenia. PMID- 15545331 TI - Homocysteine and B vitamins relate to brain volume and white-matter changes in geriatric patients with psychiatric disorders. AB - OBJECTIVE: There is a growing literature on the relationship between low serum B vitamins, elevated homocysteine, and cognitive impairment; however, few studies have examined radiological markers of associated neuropathology in geropsychiatry inpatients. The authors examined the relationship of homocysteine, folate, and vitamin B12 with magnetic resonance imaging (MRI) markers of neuropathology. METHODS: In this archival study, authors reviewed the MRIs and medical records of 34 inpatients in a geriatric psychiatry unit. Patients were selected if folate, B12, and/or homocysteine levels had been assessed and if the appropriate clinical MRIs were performed (19 men; mean age, 75 years). Patients with schizophrenia or current substance dependence were excluded. The relationships between MRI volume measures, white-matter hyperintensity (WMH) grade, and serum concentrations of folate, B12, and homocysteine were analyzed, using age-adjusted Pearson correlations. RESULTS: Homocysteine was related to WMH grade, but not brain volume measures. Folate was associated with hippocampus and amygdala, and negatively associated with WMH. B12 level was not statistically associated with any brain measure. CONCLUSIONS: Elevated homocysteine and low folate were associated with radiological markers of neuropathology. Since no patient had clinically deficient folate, it may be important to rethink what defines functionally significant micronutrient deficiency and explore what this means in different age- and health-status groups. Larger samples will be needed to assess interactions between homocysteine, micronutrients, and other neuropathology risk factors. PMID- 15545332 TI - Cognitive impairment in elderly medical inpatients: detection and associated six month outcomes. AB - OBJECTIVE: The authors examined the relationship of cognitive impairment at hospital admission to 6-month outcome (hospital readmission, nursing home admission, and death) in a cohort of elderly medical inpatients. METHODS: A group of 401 medical inpatients age 75 and older underwent a comprehensive geriatric assessment at hospital admission and were followed up for 6 months. Cognitive impairment was defined as a score <24 on the Mini-Mental State Exam. Detection was assessed through blinded review of discharge summary. Follow-up data were gathered from the centralized billing system (hospital and nursing home admissions) and from proxies (death). RESULTS: Cognitive impairment was present in 129 patients (32.3%). Only 48 (37.2%) were detected; these had more severe impairment than undetected cases. During follow-up, cognitive impairment, whether detected or not, was associated with death and nursing home admission. After adjustment for health, functional, and socioeconomic status, an independent association remained only for nursing home admission in subjects with detected impairment. Those with undetected impairment appeared to be at intermediate risk, but this relationship was not statistically significant. CONCLUSION: In these elderly medical inpatients, cognitive impairment was frequent, rarely detected, and associated with nursing home admission during follow-up. Although this association was stronger in those with detected impairment, these results support the view that acute hospitalization presents an opportunity to better detect cognitive impairment in elderly patients and target further interventions to prevent adverse outcomes such as nursing home admission. PMID- 15545333 TI - Neural basis for the cognitive continuum in episodic memory from health to Alzheimer disease. AB - OBJECTIVE: The authors hypothesize that, behaviorally, episodic memory in health and disease reflects a continuum. METHODS: Subjects (N=12) with very mild Alzheimer disease (AD) and normal subjects (N=24) were investigated with functional magnetic resonance imaging (fMRI) during an episodic memory task. RESULTS: Recruitment of a posterior medio-temporal network was correlated with memory performance across the spectrum from high- and low-performing normal subjects to patients with early AD. CONCLUSIONS: The behavioral spectrum from health to disease in episodic memory function is mirrored neurobiologically with graded recruitment of neuronal activation in medio-temporal regions. The results call for longitudinal assessment of behavioral decline and neuronal recruitment in future studies. PMID- 15545334 TI - Sex differences in brain structure in geriatric depression. AB - OBJECTIVE: In an exploratory study, authors compared patients with late-life major depression (MDD) and age-matched control subjects to examine sex differences in frontal and orbito-frontal (OFC) regional brain volumes. METHODS: The study sample comprised 41 patients with MDD and 41 controls. Subjects underwent comprehensive neuropsychiatric examinations and magnetic resonance imaging (MRI) scans. Corrected frontal and OFC gray-matter volumes were compared among men and women in both groups. RESULTS: The depression group had lower MMSE scores, greater severity of medical burden, apathy, psychomotor retardation, and poor health-related quality of life than the controls. Men in both groups had a greater severity of medical burden, apathy, and psychomotor retardation than women. The depression group had smaller OFC total and gray-matter volumes than the controls after matching for age. Men had smaller frontal-matter volumes than women in both groups. The diagnosis x sex interaction in brain regional volumes was observed only after controlling for medical burden. CONCLUSIONS: Sex differences in brain neuroanatomy may be important in the pathophysiology of geriatric depression. Men may be susceptible to atrophy in frontal subregions. Medical burden may contribute to the diagnosis x sex interaction in brain regional volumes. PMID- 15545335 TI - Comment on Nasrallah et al: lower mortality in geriatric patients receiving risperidone and olanzapine versus haloperidol. PMID- 15545336 TI - SPECT study in oldest old Alzheimer disease patients. PMID- 15545341 TI - In the beginning, there was the cell: cellular homeostasis. AB - In the past 5 years, the biomedical, scientific community has sequenced the genomes of several organisms (including Homo sapiens), has cloned entire organisms and has determine the molecular structures for several membrane proteins. These advances combined with the advances in technology enabling high throughput drug screening, gene expression readout using DNA chips and evolving proteomic techniques, make it imperative that physiologist and biomedical professionals understand the basis of cellular function and homeostasis. The Cellular Homeostasis Refresher Course at Experimental Biology 2004 in Washington, DC, was designed to fulfill this need. The specific topics covered were 1) generation of membrane potential, 2) an update on cellular mechanisms of ion homeostasis, channels and transporters, and 3) cellular volume homeostasis, and regulation of intracellular pH. PMID- 15545342 TI - Generation of resting membrane potential. AB - This brief review is intended to serve as a refresher on the ideas associated with teaching students the physiological basis of the resting membrane potential. The presentation is targeted toward first-year medical students, first-year graduate students, or senior undergraduates. The emphasis is on general concepts associated with generation of the electrical potential difference that exists across the plasma membrane of every animal cell. The intention is to provide students a general view of the quantitative relationship that exists between 1) transmembrane gradients for K(+) and Na(+) and 2) the relative channel-mediated permeability of the membrane to these ions. PMID- 15545343 TI - Ion homeostasis, channels, and transporters: an update on cellular mechanisms. AB - The steady-state maintenance of highly asymmetric concentrations of the major inorganic cations and anions is a major function of both plasma membranes and the membranes of intracellular organelles. Homeostatic regulation of these ionic gradients is critical for most functions. Due to their charge, the movements of ions across biological membranes necessarily involves facilitation by intrinsic membrane transport proteins. The functional characterization and categorization of membrane transport proteins was a major focus of cell physiological research from the 1950s through the 1980s. On the basis of these functional analyses, ion transport proteins were broadly divided into two classes: channels and carrier type transporters (which include exchangers, cotransporters, and ATP-driven ion pumps). Beginning in the mid-1980s, these functional analyses of ion transport and homeostasis were complemented by the cloning of genes encoding many ion channels and transporter proteins. Comparison of the predicted primary amino acid sequences and structures of functionally similar ion transport proteins facilitated their grouping within families and superfamilies of structurally related membrane proteins. Postgenomics research in ion transport biology increasingly involves two powerful approaches. One involves elucidation of the molecular structures, at the atomic level in some cases, of model ion transport proteins. The second uses the tools of cell biology to explore the cell-specific function or subcellular localization of ion transport proteins. This review will describe how these approaches have provided new, and sometimes surprising, insights regarding four major questions in current ion transporter research. 1) What are the fundamental differences between ion channels and ion transporters? 2) How does the interaction of an ion transport protein with so-called adapter proteins affect its subcellular localization or regulation by various intracellular signal transduction pathways? 3) How does the specific lipid composition of the local membrane microenvironment modulate the function of an ion transport protein? 4) How can the basic functional properties of a ubiquitously expressed ion transport protein vary depending on the cell type in which it is expressed? PMID- 15545344 TI - Cellular volume homeostasis. AB - All cells face constant challenges to their volume either through changes in intracellular solute content or extracellular osmolality. Cells respond to volume perturbations by activating membrane transport and/or metabolic processes that result in net solute loss or gain and return of cell volume to its normal resting state. This paper provides a brief overview of fundamental concepts of osmotic water flow across cell membranes, mechanisms of cell volume perturbation, the role of inorganic ions and organic osmolytes in cell volume regulation and the signaling mechanisms that regulate the activity of cell volume-sensitive transport and metabolic pathways. PMID- 15545345 TI - Regulation of intracellular pH. AB - The approach that most animal cells employ to regulate intracellular pH (pH(i)) is not too different conceptually from the way a sophisticated system might regulate the temperature of a house. Just as the heat capacity (C) of a house minimizes sudden temperature (T) shifts caused by acute cold and heat loads, the buffering power (beta) of a cell minimizes sudden pH(i) shifts caused by acute acid and alkali loads. However, increasing C (or beta) only minimizes T (or pH(i)) changes; it does not eliminate the changes, return T (or pH(i)) to normal, or shift steady-state T (or pH(i)). Whereas a house may have a furnace to raise T, a cell generally has more than one acid-extruding transporter (which exports acid and/or imports alkali) to raise pH(i). Whereas an air conditioner lowers T, a cell generally has more than one acid-loading transporter to lower pH(i). Just as a house might respond to graded decreases (or increases) in T by producing graded increases in heat (or cold) output, cells respond to graded decreases (or increases) in pH(i) with graded increases (or decreases) in acid-extrusion (or acid-loading) rate. Steady-state T (or pH(i)) can change only in response to a change in chronic cold (or acid) loading or chronic heat (or alkali) loading as produced, for example, by a change in environmental T (or pH) or a change in the kinetics of the furnace (or acid extrudes) or air conditioner (or acid loaders). Finally, just as a temperature-control system might benefit from environmental sensors that provide clues about cold and heat loading, at least some cells seem to have extracellular CO(2) or extracellular HCO(3)(-) sensors that modulate acid base transport. PMID- 15545346 TI - Homeostasis: a plea for a unified approach. AB - Accompanying the progressive erosion of a coherent sense of physiology as an intellectual discipline, there has been a tendency to lose sight of the homeostatic principles that underpin physiological science, and to teach them in an oversimplified form. When (as is increasingly the case) these principles are rediscovered, they are often treated as something both novel and distinct from homeostasis, fragmenting what is best understood and taught as a unified whole. This article urges a more unitary approach to homeostasis, and attempts to show how such an approach can be presented. PMID- 15545347 TI - Non-science majors gain valuable insight studying clinical trials literature: an evidence-based medicine library assignment. AB - When faced with a diagnosis, it is empowering to be able to assess the evidence of treatment effectiveness and safety. To teach this skill to non-science majors, we assigned the "Responsible Patienthood Project" (RPP). For the RPP, students studied an array of disease and treatment literature: the final product of their work was a poster presentation, in which they did an in-depth analysis of one primary article, thus encouraging critical evaluation of experimental design, methods, and conclusions. Post-RPP, there was a 35% decrease in the student perception that they would unquestioningly accept a recommended treatment for a hypothetical diagnosis, and a 40% increase in the perception that they would consult a combination of resources, including primary articles. We recommend this project based on our results that suggest 1) non-science majors are able to successfully access and assess primary scientific literature, 2) students felt empowered by the RPP, and 3) skills in information gathering, via library instruction, may serve as a particularly helpful lifelong learning tool. PMID- 15545348 TI - Evaluating metabolic syndrome in a medical physiology laboratory. AB - The metabolic syndrome, a cluster of factors linked to obesity that contribute to risk for atherosclerosis and Type 2 diabetes, may affect 20-25% of the adults in the United States. We designed a medical physiology laboratory to evaluate and discuss the physiological and nutritional principles involved in the metabolic syndrome. The five criteria used to diagnose this syndrome (fasting blood triglycerides, high-density lipoprotein cholesterol, and glucose, blood pressure, central obesity) were measured by students on each other either previously or during this exercise. In addition, to illustrate nutritional factors involved in causation and treatment of the metabolic syndrome, a meal was provided during the laboratory. Class members were randomized to groups allowed ad libitum meal composition, or constrained to the National Cholesterol Education Program Step I or Step II diets. The composition of the diet (including saturated fat, cholesterol, dietary fiber, and carbohydrate content) was discussed in the context of blood cholesterol, triglyceride, and glucose levels. This laboratory allows a comprehensive analysis of the physiological and nutritional factors involved in the development of the metabolic syndrome. PMID- 15545349 TI - Infusion of collaborative inquiry throughout a biology curriculum increases student learning: a four-year study of "Teams and Streams". AB - Are traditional laboratories in the core introductory biology courses teaching physiology majors the art and trade of science, or simply leaving them with a memory of trivial experiments done for unknown reasons? Our students, a population dominated by pre-med and physiology majors, think the latter and have encouraged us to challenge this model, and it turns out scientists and education researchers agree with our students (4, 31, 32). In an effort to remedy this, we began a long-term redesign of the introductory biology sequence to become what is now a sequence of inquiry laboratories we term "Teams and Streams" (TS). In these TS inquiry labs, student research teams pose a scientific question/hypothesis, propose an experimental design, perform multi-week investigations and then present their findings in various forms (web, interviews, and papers). The response to this classroom laboratory design has been overwhelmingly positive. In a qualitative study of student opinion (where 260 student responses were studied), surveys conducted at the end of semesters where traditional scripted labs were used (n = 70 comments) had predominantly negative opinions (80% negative responses), whereas the reverse was true for students (n = 190 comments) who participated in courses using the TS inquiry labs (78% positive responses). In a quantitative assessment of content knowledge, students who participated in new TS inquiry labs (n = 245) outscored their peers in traditional labs (n = 86) on Medical College Admission Test-style standardized exams (59.3 +/- 0.8% vs. 48.9 +/- 1.3%, respectively; P < 0.0001). We believe these quantitative data support the qualitative findings and suggest the TS inquiry lab approach increases student learning. PMID- 15545350 TI - Addressing students' misconceptions of renal clearance. AB - Renal clearance is one of the more difficult concepts for students of physiology to learn. We hypothesized that this difficulty is rooted in a student's misunderstanding of virtual volume. This was tested by having students select from several drawings the one they thought plasma would look like after a certain volume of it has been cleared of sodium by the kidneys. About half the participating students selected plasma pictured as having a certain volume of it devoid of sodium molecules. That is, their misconception of clearance seemed to be due to a lack of understanding about virtual volume, a deficiency which is reinforced by the classic definition of clearance. To address this misconception, a demonstration was devised in which a beaker of concentrated colored water was used to represent plasma before renal clearance, a beaker of the same concentrated colored water in which the top third had been replaced by clear mineral oil was used to represent what the definition of clearance said would happen to plasma after a third of it had been cleared of sodium, and a beaker of dilute colored water was used to represent what really happens to plasma when a certain volume of it is cleared of a solute. Incorporating this demonstration into discussions of renal clearance helped students to understand this concept, as evidenced by improved scores on related questions. PMID- 15545351 TI - The targeted oral. AB - An oral examination for assessing senior undergraduate students is described. The examinations were conducted in a room with a one-way mirror so that all students could learn from each other. This procedure was much appreciated by the participants. PMID- 15545352 TI - Determinants of human immunodeficiency virus type 1 escape from the primary CD8+ cytotoxic T lymphocyte response. AB - CD8+ cytotoxic T lymphocytes (CTLs) play an important role in containment of virus replication in primary human immunodeficiency virus (HIV) infection. HIV's ability to mutate to escape from CTL pressure is increasingly recognized; but comprehensive studies of escape from the CD8 T cell response in primary HIV infection are currently lacking. Here, we have fully characterized the primary CTL response to autologous virus Env, Gag, and Tat proteins in three patients, and investigated the extent, kinetics, and mechanisms of viral escape from epitope-specific components of the response. In all three individuals, we observed variation beginning within weeks of infection at epitope-containing sites in the viral quasispecies, which conferred escape by mechanisms including altered peptide presentation/recognition and altered antigen processing. The number of epitope-containing regions exhibiting evidence of early CTL escape ranged from 1 out of 21 in a subject who controlled viral replication effectively to 5 out of 7 in a subject who did not. Evaluation of the extent and kinetics of HIV-1 escape from >40 different epitope-specific CD8 T cell responses enabled analysis of factors determining escape and suggested that escape is restricted by costs to intrinsic viral fitness and by broad, codominant distribution of CTL mediated pressure on viral replication. PMID- 15545353 TI - Absence of caspase 8 and high expression of PED protect primitive neural cells from cell death. AB - The mechanisms that control neural stem and progenitor cell survival are unknown. In several pathological conditions, death receptor (DR) ligands and inflammatory cytokines exert a deleterious effect on neurons, whereas primitive neural cells migrate and survive in the site of lesion. Here, we show that even in the presence of inflammatory cytokines, DRs are unable to generate death signals in primitive neural cells. Neural stem and progenitor cells did not express caspase 8, the presence of which is required for initiating the caspase cascade. However, exogenous or cytokine-mediated expression of caspase 8 was not sufficient to restore their DR sensitivity. Searching for molecules potentially able to block DR death-inducing signaling complex (DISC), we found that primitive neural cells expressed high levels of the death effector domain-containing protein PED (also known as PEA-15). PED localized in the DISC and prevented caspase 8 recruitment and activation. Moreover, lentiviral-mediated delivery of PED antisense DNA resulted in dramatic down-regulation of the endogenous gene expression and sensitization of primitive neural cells to apoptosis mediated by inflammatory cytokines and DRs. Thus, absence of caspase 8 and high expression of PED constitute two levels of protection from apoptosis induced by DRs and inflammatory cytokines in neural stem and progenitor cells. PMID- 15545354 TI - DC-SIGN-mediated infectious synapse formation enhances X4 HIV-1 transmission from dendritic cells to T cells. AB - Dendritic cells (DCs) are essential for the early events of human immunodeficiency virus (HIV) infection. Model systems of HIV sexual transmission have shown that DCs expressing the DC-specific C-type lectin DC-SIGN capture and internalize HIV at mucosal surfaces and efficiently transfer HIV to CD4+ T cells in lymph nodes, where viral replication occurs. Upon DC-T cell clustering, internalized HIV accumulates on the DC side at the contact zone (infectious synapse), between DCs and T cells, whereas HIV receptors and coreceptors are enriched on the T cell side. Viral concentration at the infectious synapse may explain, at least in part, why DC transmission of HIV to T cells is so efficient.Here, we have investigated the role of DC-SIGN on primary DCs in X4 HIV 1 capture and transmission using small interfering RNA-expressing lentiviral vectors to specifically knockdown DC-SIGN. We demonstrate that DC-SIGN- DCs internalize X4 HIV-1 as well as DC-SIGN+ DCs, although binding of virions is reduced. Strikingly, DC-SIGN knockdown in DCs selectively impairs infectious synapse formation between DCs and resting CD4+ T cells, but does not prevent the formation of DC-T cells conjugates. Our results demonstrate that DC-SIGN is required downstream from viral capture for the formation of the infectious synapse between DCs and T cells. These findings provide a novel explanation for the role of DC-SIGN in the transfer and enhancement of HIV infection from DCs to T cells, a crucial step for HIV transmission and pathogenesis. PMID- 15545355 TI - Insufficient production and tissue delivery of CD4+ memory T cells in rapidly progressive simian immunodeficiency virus infection. AB - The mechanisms linking human immunodeficiency virus replication to the progressive immunodeficiency of acquired immune deficiency syndrome are controversial, particularly the relative contribution of CD4+ T cell destruction. Here, we used the simian immunodeficiency virus (SIV) model to investigate the relationship between systemic CD4+ T cell dynamics and rapid disease progression. Of 18 rhesus macaques (RMs) infected with CCR5-tropic SIVmac239 (n=14) or CXCR4 tropic SIVmac155T3 (n=4), 4 of the former group manifested end-stage SIV disease by 200 d after infection. In SIVmac155T3 infections, naive CD4+ T cells were dramatically depleted, but this population was spared by SIVmac239, even in rapid progressors. In contrast, all SIVmac239-infected RMs demonstrated substantial systemic depletion of CD4+ memory T cells by day 28 after infection. Surprisingly, the extent of CD4+ memory T cell depletion was not, by itself, a strong predictor of rapid progression. However, in all RMs destined for stable infection, this depletion was countered by a striking increase in production of short-lived CD4+ memory T cells, many of which rapidly migrated to tissue. In all rapid progressors (P <0.0001), production of these cells initiated but failed by day 42 of infection, and tissue delivery of new CD4+ memory T cells ceased. Thus, although profound depletion of tissue CD4+ memory T cells appeared to be a prerequisite for early pathogenesis, it was the inability to respond to this depletion with sustained production of tissue-homing CD4+ memory T cells that best distinguished rapid progressors, suggesting that mechanisms of the CD4+ memory T cell generation play a crucial role in maintaining immune homeostasis in stable SIV infection. PMID- 15545356 TI - NKG2D recognition and perforin effector function mediate effective cytokine immunotherapy of cancer. AB - Single and combination cytokines offer promise in some patients with advanced cancer. Many spontaneous and experimental cancers naturally express ligands for the lectin-like type-2 transmembrane stimulatory NKG2D immunoreceptor; however, the role this tumor recognition pathway plays in immunotherapy has not been explored to date. Here, we show that natural expression of NKG2D ligands on tumors provides an effective target for some cytokine-stimulated NK cells to recognize and suppress tumor metastases. In particular, interleukin (IL)-2 or IL 12 suppressed tumor metastases largely via NKG2D ligand recognition and perforin mediated cytotoxicity. By contrast, IL-18 required tumor sensitivity to Fas ligand (FasL) and surprisingly did not depend on the NKG2D-NKG2D ligand pathway. A combination of IL-2 and IL-18 stimulated both perforin and FasL effector mechanisms with very potent effects. Cytokines that stimulated perforin-mediated cytotoxicity appeared relatively more effective against tumor metastases expressing NKG2D ligands. These findings indicate that a rational choice of cytokines can be made given the known sensitivity of tumor cells to perforin, FasL, and tumor necrosis factor-related apoptosis-inducing ligand and the NKG2D ligand status of tumor metastases. PMID- 15545357 TI - Secretory leukocyte protease inhibitor binds to annexin II, a cofactor for macrophage HIV-1 infection. AB - The distribution of secretory leukocyte protease inhibitor (SLPI) at entry portals indicates its involvement in defending the host from pathogens, consistent with the ability of SLPI to inhibit human immunodeficiency virus (HIV) 1 infection by an unknown mechanism. We now demonstrate that SLPI binds to the membrane of human macrophages through the phospholipid-binding protein, annexin II. Based on the recent identification of human cell membrane phosphatidylserine (PS) in the outer coat of HIV-1, we define a novel role for annexin II, a PS binding moiety, as a cellular cofactor supporting macrophage HIV-1 infection. Moreover, this HIV-1 PS interaction with annexin II can be disrupted by SLPI or other annexin II-specific inhibitors. The PS-annexin II connection may represent a new target to prevent HIV-1 infection. PMID- 15545358 TI - Age-related CD8 T cell clonal expansions constrict CD8 T cell repertoire and have the potential to impair immune defense. AB - Peripheral T cell diversity is virtually constant in the young, but is invariably reduced in aged mice and humans. CD8+ T cell clonal expansions (TCE) are the most drastic manifestation of, and possible contributors to, this reduced diversity. We show that the presence of TCE results in reduced CD8+, but not CD4+, T cell diversity, and in functional inability to mobilize parts of the CD8+ T cell repertoire affected by TCE. In the model of herpes simplex virus (HSV)-1 infection of B6 mice, >90% of the responding CD8+ T cells use Vbeta10 or Vbeta8 and are directed against a single glycoprotein B (gB498-505) epitope, gB-8p. We found that old animals bearing CD8+ TCE within Vbeta10 or Vbeta8 families failed to mount an effective immune response against HSV-1, as judged by reduced numbers of peptide-major histocompatibility complex tetramer+ CD8 T cells and an absence of antiviral lytic function. Furthermore, Vbeta8 TCE experimentally introduced into young mice resulted in lower resistance to viral challenge, whereas Vbeta5+ TCE induced in a similar fashion did not impact viral resistance. These results demonstrate that age-related TCE functionally impair the efficacy of antiviral CD8+ T cell immunity in an antigen-specific manner, strongly suggesting that TCE are not the mere manifestation of, but are also a contributing factor to, the immunodeficiency of senescence. PMID- 15545359 TI - Short communication: utilization of sheep's milk cheese whey in the manufacture of an alkylphenol flavor concentrate. AB - The recovery of species-related conjugated sheep-like flavored alkylphenols from Manchego-type cheese whey by ultrafiltration was investigated. Concentrations of conjugated alkylphenols were similar in the various fractions of whey permeate collected during ultrafiltration, and this was interpreted as a reflection of their high water solubility. About 49 and 62% of conjugated 3- and 4-ethylphenols and p- and m-cresols in sheep's milk cheese whey, respectively, were recovered in the permeate after ultrafiltration with a volume concentration factor of 5.4. Cheese whey retentate correspondingly contained 38 and 28% of conjugated 3- and 4 ethylphenols and p- and m-cresols from the original whey, respectively. Permeate fractions from sheep's milk cheese whey were combined, concentrated by vacuum evaporation, and lactose was partially removed by crystallization and filtration to obtain an aqueous sheep-like flavor precursor concentrate. PMID- 15545360 TI - Comparison of effect of vacuum-condensed and ultrafiltered milk on cheddar cheese. AB - The objective of this study was to compare the effects of vacuum-condensed (CM) and ultrafiltered (UF) milk on some compositional and functional properties of Cheddar cheese. Five treatments were designed to have 2 levels of concentration (4.5 and 6.0% protein) from vacuum-condensed milk (CM1 and CM2) and ultrafiltered milk (UF1 and UF2) along with a 3.2% protein control. The samples were analyzed for fat, protein, ash, calcium, and salt contents at 1 wk. Moisture content, soluble protein, meltability, sodium dodecyl sulfate-PAGE, and counts of lactic acid bacteria and nonstarter lactic acid bacteria were performed on samples at 1, 18, and 30 wk. At 1 wk, the moisture content ranged from 39.2 (control) to 36.5% (UF2). Fat content ranged from 31.5 to 32.4% with no significant differences among treatments, and salt content ranged from 1.38 to 1.83% with significant differences. Calcium content was higher in UF cheeses than in CM cheeses followed by control, and it increased with protein content in cheese milk. Ultrafiltered milk produced cheese with higher protein content than CM milk. The soluble protein content of all cheeses increased during 30 wk of ripening. Condensed milk cheeses exhibited a higher level of proteolysis than UF cheeses. Sodium dodecyl sulfate-PAGE showed retarded proteolysis with increase in level of concentration. The breakdown of alphas1- casein and alphas1-I-casein fractions was highest in the control and decreased with increase in protein content of cheese milk, with UF2 being the lowest. There was no significant degradation of beta-casein. Overall increase in proteolytic products was the highest in control, and it decreased with increase in protein content of cheese milk. No significant differences in the counts of lactic starters or nonstarter lactic acid bacteria were observed. Extent as well as method of concentration influenced the melting characteristics of the cheeses. Melting was greatest in the control cheeses and least in cheese made from condensed milk and decreased with increasing level of milk protein concentration. Vacuum condensing and ultrafiltration resulted in Cheddar cheeses of distinctly different quality. Although both methods have their advantages and disadvantages, the selection of the right method would depend upon the objective of the manufacturer and intended use of the cheese. PMID- 15545361 TI - High-pressure-induced interactions between milk fat globule membrane proteins and skim milk proteins in whole milk. AB - The association of beta-lactoglobulin (beta-LG) and alpha-lactalbumin (alpha-LA) with milk fat globule membrane (MFGM), when whole milk was treated by high pressure in the range 100 to 800 MPa, was investigated using sodium dodecyl sulfate (SDS)-PAGE under reducing and nonreducing conditions. In SDS-PAGE under reducing conditions, beta-LG was observed in the MFGM material isolated from milk treated at 100 to 800 MPa for 30 min, and small amounts of alpha-LA and kappa casein were also observed at pressures >600 MPa for 30 min. However, these proteins were not observed in SDS-PAGE under nonreducing conditions. These results indicate that beta-LG and alpha-LA associated with MFGM proteins via disulfide bonds during the high-pressure treatment of whole milk. The amount of beta-LG associated with the MFGM increased with an increase in pressure up to 800 MPa and with increasing time of pressure treatment. The maximum value for beta-LG association with the MFGM was approximately 0.75 mg/g of fat. Of the major original MFGM proteins, no change in butyrophilin was observed during the high pressure treatment of whole milk, whereas xanthine oxidase was reduced to some extent beyond 400 MPa. In contrast to the behavior during heat treatment, PAS 6 and PAS 7 were stable during high-pressure treatment, and they remained associated with the MFGM. PMID- 15545362 TI - Effect of genetic variation on the tryptic hydrolysis of bovine beta lactoglobulin A, B, and C. AB - The structure, stability, and hydrolysis characteristics of beta-lactoglobulin (LG) A are different from those of either beta-LG B or beta-LG C. Purified samples of these proteins were hydrolyzed with trypsin and the rates of loss of native monomeric beta-LG structure were measured using sodium dodecyl sulfate PAGE. At the same time, the appearance of many individual peptides were identified and followed in time by HPLC, measuring their concentration as a function of solution pH, temperature, protein concentration, and added urea or palmitate. The identity of the peptides was confirmed by liquid chromatography mass spectrometry. This semiquantitative exploration showed that the rate of hydrolysis was in the order beta-LG A > beta-LG B > beta-LG C under most circumstances, and that 12 of the 18 trypsin-susceptible bonds were cleaved at very similar rates that were governed by the variant type. Consequently, the rate of hydrolysis of the intact protein was related to the overall structural stability of the individual proteins and the accessibility of certain peptide bonds to the enzyme. The hydrolysis of mixtures of 2 or more variants or of denatured beta-LG gave more heterogeneous peptide mixtures. PMID- 15545363 TI - Textural and sensory characteristics of whole and skimmed flavored set-type yogurt during long storage. AB - A study of refrigerated storage (10 degrees C for 91 d) of whole and skimmed flavored set-type yogurt was made. Comparison with storage at 20 degrees C for 21 d and 30 degrees C for 3 d (accelerated) was also carried out. Refrigerated storage yogurts were assessed by a trained panel and by a consumer panel. Trained panel scores were correlated to instrumental data, and the acceptability data for long storage were studied using consumer criteria. In all cases, after-storage pH values barely changed over storage time, indicating that the yogurt samples did not develop much acidity under any of the storage conditions studied. The profile of the instrumental texture curves obtained corresponded to a firm gel, which broke after a plunger penetrated the sample, and the firmness values of the whole yogurt were lower than for the skimmed yogurt under all the storage conditions studied. From a microbiological point of view, the viability of the yogurts was adequate at the different storage times and temperatures studied, although those stored at 10 degrees C for long periods would not comply with some countries' minimum requirements. Logistic regression of the data from a 50-consumer sensory evaluation showed that the probability of the whole yogurt being accepted after 91 d storage at 10 degrees C was around 40%, whereas for the skimmed yogurt it was only 15%, largely because the skimmed yogurt developed certain negative attributes at an earlier stage of storage than the whole yogurt. PMID- 15545364 TI - Mathematical models for combined high pressure and thermal plasmin inactivation kinetics in two model systems. AB - The combined high-pressure thermal inactivation kinetics of plasmin was studied in 2 model systems. The first system contained both plasmin and plasminogen, whereas, in the second system, all plasminogen was converted into plasmin, with urokinase, before the inactivation studies. High-pressure treatments were conducted in the range of 300 to 800 MPa combined with temperatures from 30 to 65 degrees C. Under all conditions of pressure and temperature (isobaric-isothermal) studied, for both systems, first-order inactivation was observed. A third-degree polynomial model (derived from thermodynamic principles) successfully described the temperature and pressure dependence of the inactivation rate constant over the entire experimental domain. The antagonistic effect and the stabilization effect observed above a threshold pressure value of 600 MPa were thought to be related to the disruption of disulfide bonds in plasmin and plasminogen. PMID- 15545365 TI - Screening of dairy yeast strains for probiotic applications. AB - To evaluate the potential of yeasts of dairy origin as probiotics, we tested 8 species including Candida humilis, Debaryomyces hansenii, Debaryomyces occidentalis, Kluyveromyces lactis, Kluyveromyces lodderae, Kluyveromyces marxianus, Saccharomyces cerevisiae, and Yarrowia lipolytica, isolated from commercial blue cheese and kefir. Strains were randomly selected from each species and tested for their ability to adhere to human enterocyte-like Caco-2 cells in culture. Among the 8 species, K. lactis showed higher adhesive ability than K. marxianus, K. lodderae, and D. hansenii. The other 4 species were poorly adhesive. All species other than K. marxianus and C. humilis were resistant to acidic conditions. In the presence of bile acid, growth inhibition was undetectable when incubation was carried out at 27 degrees C; however, it was evident for C. humilis and a strain of D. occidentalis when incubated at 37 degrees C. Moreover, the influence of proteinase treatment of living cells of K. lactis and K. lodderae on their adhesion to Caco-2 cells was evaluated. Although a slight reduction was recognized when K. lactis was treated with proteinase K, the influence of intestinal protease treatments of pepsin followed by trypsin was negligible. These results indicated that a proteinaceous factor was unlikely to be involved in adhesion of K. lactis and K. lodderae to Caco-2 cells. No stimulation of IL-8 synthesis by Caco-2 cells was recognized in the presence of K. lactis. In conclusion, K. lactis was the most attractive to continue study for use as probiotic microorganisms. PMID- 15545366 TI - Aroma compounds in sweet whey powder. AB - Aroma compounds in sweet whey powder were investigated in this study. Volatiles were isolated by solvent extraction followed by solvent-assisted flavor evaporation. Fractionation was used to separate acidic from nonacidic volatiles. Gas chromatography/mass spectrometry and gas chromatography/olfactometry were used for the identification of aroma compounds. Osme methodology was applied to assess the relative importance of each aroma compound. The most aroma-intense free fatty acids detected were acetic, propanoic, butanoic, hexanoic, heptanoic, octanoic, decanoic, dodecanoic, and 9-decenoic acids. The most aroma-intense nonacidic compounds detected were hexanal, heptanal, nonanal, phenylacetaldehyde, 1-octen-3-one, methional, 2,6-dimethylpyrazine, 2,5-dimethylpyrazine, 2,3 dimethylpyrazine, 2,3,5-trimethylpyrazine, furfuryl alcohol, p-cresol, 2 acetylpyrrole, maltol, furaneol, and several lactones. This study suggested that the aroma of whey powder could comprise compounds originating from milk, compounds generated by the starter culture during cheese making, and compounds formed during the manufacturing process of whey powder. PMID- 15545367 TI - Volatile free fatty acids as ripening indicators for Serra da Estrela cheese. AB - Serra da Estrela cheese is an artisanal cheese manufactured from raw ewe's milk coagulated with Cynara cardunculus; it is highly appreciated for its unique flavor and bouquet. This research effort focused on a search for the molecules responsible for those organoleptic characteristics. Eighty cheeses manufactured in 4 dairies located in the Appelation d'Origine Protegee region of Serra da Estrela, Portugal, were thus assayed for volatiles, in an attempt to characterize their odorous profile. Cheeses were analyzed from the time of manufacture up to 180 d of ripening. The volatile fraction was assayed by solid phase microextraction-gas chromatography/mass spectroscopy, and several compounds belonging to different chemical groups (e.g., fatty acids, esters, carbonyl compounds, pyrazines, and sulfur compounds) were detected in that traditional cheese. Among these, free fatty acids (FFA) were quantitatively the dominant family present. Furthermore, sensory descriptors for the typical aroma of this cheese included acidic, sweaty, and sheepy-like. Acetic, isobutyric, and isovaleric acids increased in concentration during the ripening process up to 90 d, and remained constant thereafter. Preliminary sensory analysis was performed by addition of the major FFA to an unripened cheese matrix; results showed that they could successfully be used as ripening indicators for this cheese. Such key molecules may thus be used to monitor ripening, and hence find the optimal consumption time for this gourmet dairy product. PMID- 15545368 TI - Suppressive effect of functional drinking yogurt containing specific egg yolk immunoglobulin on Helicobacter pylori in humans. AB - Helicobacter pylori is a human pathogen that infects over 50% of the population worldwide. It is the most important etiologic agent of gastroduodenal ulcers and malignancies. Helicobacter pylori urease enzyme is considered the main factor for the organism's colonization in the gastroduodenal mucosa. Hens immunized with the purified urease produce a highly specific anti-H. pylori urease immunoglobulin (IgY-urease) in their egg yolks. Immunoglobulin Y-urease was stable at 60 to 65 degrees C for 30 min and at pH 4.0 for 7 h. Its activity was lost at 80 degrees C for 20 min and at pH 2 for 4 h. Specially designed functional drinking yogurt containing Lactobacillus acidophilus and Bifidobacterium spp. with 1% egg yolk IgY-urease was produced commercially. Immunoglobulin Y-urease activity showed stability in the product up to 7 d, and then decreased to 85% after 3 wk of storage. A clinical study was conducted to determine the effectiveness of IgY urease yogurt to suppress infection in humans. Forty-two volunteers who tested positive for H. pylori using a 13C-urea breath test were recruited. A total of 450 mL of IgY-urease (test group) or IgY-urease-free yogurt (control group) was consumed in 150-mL portions 3 times daily for 4 wk. Volunteers were tested after 2 and 4 wk; urea breath test values significantly decreased in the test group compared with the control group. The results indicate that suppression of H. pylori infection in humans could be achieved by consumption of drinking yogurt fortified with IgY-urease. PMID- 15545369 TI - Emulsifying properties of fractions prepared from commercial buttermilk by microfiltration. AB - A novel method for the separation of milk-fat globule membrane (MFGM) isolate by microfiltration in the presence of citrate was applied to prepare a fraction to be used to stabilize oil-in-water emulsions. The emulsifying properties of this fraction, containing high amounts of MFGM, were compared with a buttermilk concentrate (BMC) prepared in a similar manner but still containing the original ratio of proteins (caseins, whey proteins, and MFGM). The objective of this work was to determine if the isolation procedure would result in an ingredient with different functionality when compared with BMC. These fractions were incorporated into oil-in-water emulsions at various isolate and oil concentrations. At low concentrations of isolate, MFGM emulsions showed better creaming stability and smaller oil droplet size distribution than whole buttermilk concentrate samples. The difference in stability was attributed to the compositional difference between the 2 ingredients prepared. A selective concentration of MFGM in buttermilk by microfiltration has the potential for the development of ingredients that differ substantially from the ingredients deriving from milk or whey. PMID- 15545370 TI - Inhibitory effects of human and bovine milk constituents on rotavirus infections. AB - Among etiologic agents, rotavirus is the major cause of severe dehydration diarrhea in infant mammals. In vitro and in vivo studies have indicated that the human milk-fat globule protein lactadherin inhibits rotavirus binding and protects breast-fed children against symptomatic rotavirus infection. The present work was conducted to evaluate the effect of lactadherin, along with some other milk proteins and fractions, on rotavirus infections in MA104 and Caco-2 cell lines. It is shown that human, and not bovine, lactadherin inhibits Wa rotavirus infection in vitro. Human lactadherin seems to act through a mechanism involving protein-virus interactions. The reason for the activity of human lactadherin is not clear, but it might lie within differences in the protein structure or the attached oligosaccharides. Likewise, in our hands, bovine lactoferrin did not show any suppressive activity against rotavirus. In contrast, MUC1 from bovine milk inhibits the neuraminidase-sensitive rotavirus RRV strain efficiently, whereas it has no effect on the neuraminidase-resistant Wa strain. Finally, a bovine macromolecular whey protein fraction turned out to have an efficient and versatile inhibitory activity against rotavirus. PMID- 15545371 TI - Purification and characterization of intracellular proteinase from Lactobacillus casei ssp. casei LLG. AB - The intracellular proteinase of Lactobacillus casei ssp. casei LLG was isolated in the cytoplasmic fraction with 0.05 M Tris-HCl buffer (pH 7.5). The enzyme was purified by the fast protein liquid chromatography system equipped with ion exchange and gel filtration chromatographies. This proteinase comprised a single monomeric form and had a molecular weight of about 55 kDa and an isoelectric point near pH 4.9. The optimum pH and temperature for the enzyme activity were determined to be pH 6.5 and 37 degrees C, respectively. The enzyme was inactivated by metal-chelating compounds (EDTA, 1,10-phenanthroline) and less affected by serine proteinase inhibitors (diisopropylfluorophosphate, phenylmethylsulfonyl fluoride). Proteinase activity was increased by Ca++, Mn++, and Co++, and inhibited by Cu++, Mg++, and Zn++. The activity of this enzyme to hydrolyze casein appeared to be more active on beta-casein than alphas1-casein and kappa-casein as monitored by polyacrylamide gel electrophoresis. PMID- 15545372 TI - Delayed neutrophil apoptosis in bovine subclinical mastitis. AB - Bovine subclinical mastitis can be defined as a moderated inflammatory disease characterized by a persistent accumulation of neutrophils in milk. As GMCSF mediated delay of neutrophil apoptosis contributes to the accumulation of inflammatory cells at the site of inflammation in many human diseases, we sought to determine whether subclinical mastitis in cows is also associated with a GMCSF dependent increase in milk-neutrophil survival. We first addressed the hypothesis that GMCSF delays bovine neutrophil apoptosis by activation of the signal transducer and activator of transcription (STAT) family members STAT3 and STAT5, which are critical regulators of the expression of various Bcl-2 family proteins. Granulocyte-macrophage colony-stimulating factor significantly delayed apoptosis of blood neutrophils obtained from healthy cows. In these cells, GMCSF activated STAT5, but not STAT3, and induced an increase in the mRNA of the antiapoptotic Bcl-2 member, Bcl-xL. Granulocyte-macrophage colony-stimulating factor-dependent STAT5 activation and up-regulation of Bcl-xL mRNA were blocked by the Jak inhibitor, AG-490. This inhibition was associated with abrogation of the prosurvival effect of GMCSF, demonstrating a key role for STAT5 in delayed neutrophil apoptosis. We further found that GMCSF expression was increased in milk cells from cows affected with subclinical mastitis. Neutrophils from these cows demonstrated a significant delay of apoptosis as compared with neutrophils obtained from healthy cows and were unresponsive to GMCSF. Active STAT5 complexes were detected in these neutrophils. Finally, in the presence of AG-490, apoptosis was induced and a time-dependent down-regulation of Bcl-xL mRNA was observed in milk neutrophils from mastitis-affected cows. These results indicate that neutrophil survival is enhanced in milk of subclinical mastitis-affected cows and suggest a role for a GMCSF-activated STAT5 signaling pathway in this phenomenon. This pathway could thus represent a target for the control of persistent accumulation of neutrophils in the bovine mammary gland. PMID- 15545373 TI - Effect of lameness on culling in dairy cows. AB - The purpose of this study was to assess the effect of lameness on dairy cow survival. Cox's proportional hazards regression models were fitted to single lactation data from 2520 cows in 2 New York State dairy herds. Models were controlled for the time-independent effects of parity, projected milk yield, and calving season, and for the time-dependent effects of lameness and culling. Other common diseases were found to be nonconfounding and so were not included in any of the final models. Survival was measured as the time from calving until death or sale. Cows were censored if they reached the start of the next lactation or end of the study, whichever occurred first. All models were stratified by herd. For all lameness diagnoses combined, survival in the herd decreased for those cows becoming lame during the first half of lactation, with a hazard ratio of up to 2 times that of a nonlame cow. Foot rot diagnosed during the second or third months of lactation decreased survival during the same time period (hazard ratio=5.1; 95% confidence interval=1.6 to 16.2). Sole ulcers diagnosed in the first 4 mo of lactation decreased survival in several subsequent periods in which the strongest association was between diagnosis in the third and fourth months of lactation and exit from the herd during that same period (hazard ratio=2.7; 95% confidence interval=1.3 to 6.0). Foot warts were not associated with decreased survival in this analysis. Lameness was never associated with increased survival in any of the models. PMID- 15545374 TI - Effect of lameness on ovarian activity in postpartum holstein cows. AB - A longitudinal study was conducted to examine the relationship between lameness and delayed ovarian cyclicity during the first 60 d postpartum and days to first luteal activity during the first 300 d postpartum in Holstein cows. Two hundred thirty-eight cows from a 600-cow dairy that calved during a 12-mo period were used. Cows were classified into 1 of 6 categories of lameness during the first 35 d postpartum using a locomotion scoring system. Cows were blood-sampled weekly for detection of plasma progesterone concentrations during the first 300 d postpartum. Cows with delayed resumption of ovarian cyclicity were defined as those with progesterone concentrations consistently <1 ng/mL during the first 60 d postpartum. The null hypothesis that risk of delayed cyclicity is the same in cows classified as nonlame, moderately lame, or lame (after adjusting for potential modifying or confounding effects of loss of body condition and other variables related with delayed cyclicity) was tested using logistic regression. Analysis of results of the study reported here support the hypothesis that lameness is associated with delayed ovarian activity in Holstein cows during the early postpartum period. Cows classified as lame had 3.5 times greater odds of delayed cyclicity, compared with cows classified as nonlame. Attributable proportion analysis indicated that delayed ovarian cyclicity in lame cows would be reduced by 71%, if lameness had been prevented. PMID- 15545375 TI - Increase of Escherichia coli inoculum doses induces faster innate immune response in primiparous cows. AB - The objective of the current study was to evaluate the dynamics of infection and the immunological response to varying numbers of Escherichia coli injected into the mammary glands of primiparous cows during the periparturient period. Primiparous cows have been shown to be more resistant to intramammary E. coli challenge, and an increase of the inoculum dose by 2 log10 units induced a more rapid clinical response and clearance of the organisms. Recognition of lipopolysaccharide (LPS) is a key event in the innate immunity response to gram negative infection and is mediated by the accessory molecules CD14 and LPS binding protein (LBP). Primiparous cows were inoculated with 1 x 10(4) (Group A; n=8) or 1 x 10(6) (Group B; n=8) cfu E. coli P4:O32 in their 2 left quarters during the periparturient period. Clinical examination and analysis of blood and milk parameters, including IL-8, complement fragment 5a (C5a), LBP, and soluble CD14 (sCD14), were performed from d -4 to d +3 relative to infection. Primiparous cows in Group B initiated a more rapid clinical response following intramammary infection (IMI), resulting in typical clinical signs and changes in blood and milk parameters approximately 3 h earlier compared with primiparous cows in Group A. Based on average milk production in the noninfected quarters on d +2 postinoculation, all heifers reacted as moderate responders. Distinct differences in the kinetic patterns of rectal temperature, somatic cell count (SCC), IL-8, C5a, LBP, and sCD14 were observed between both groups during the early phase of inflammation. Both C5a and IL-8 increased before cellular influx into the infected glands, followed by increases in sCD14 and LBP. In conclusion, primiparous cows are able to clear an intramammary E. coli infection efficiently. Moreover, increasing the inoculum dose induces a more rapid inflammatory reaction, mainly because of early activation of the innate host immune response. PMID- 15545376 TI - Prevalence of multidrug resistant Staphylococcus aureus in milk from large- and small-scale producers in Kenya. AB - This study evaluated the prevalence of multidrug resistant Staphylococcus aureus in Kenyan milk and investigated any differences in antimicrobial resistance between large- (>200 L/d) and small- (<50 L/d) scale producers. Susceptibility profiles for penicillin G, tetracycline, erythromycin, trimethoprim/sulfamethazine, and chloramphenicol were determined for Staph. aureus (n=402) isolated from cows with subclinical mastitis. There was a significant difference in the overall mean resistance profile between large- (7.1%) and small-scale farm (14.7%) isolates. The overall prevalence of multidrug resistance (> or =2 antibiotics) differed significantly between isolates from small farms (34.3%) and those from large farms (18.0%). Additionally, the producers were interviewed about their usage of antimicrobial drugs and their attitudes toward education in related fields. There was an evident difference between the producer types in their documentation of the use of antimicrobial drugs. Small-scale farms were less inclined to documentation, and treatment records were available from 22% of small-scale farms, compared with 73% of large scale farms. Farmers expressed a need for more information in 5 areas, ranking preventive management highest (34.0%); followed by affordable tests to control residues in milk (22.8%); preparation of antimicrobial drugs (20.0%); public health concerns (11.2%); disposal of surplus antimicrobial drugs (7.8%); and antimicrobial drug residue persistence in milk (4.2%). It was concluded that herd size might be an indirect risk factor in the development of antimicrobial resistance in Staph. aureus within the region. PMID- 15545377 TI - Viability of milk neutrophils and severity of bovine coliform mastitis. AB - To study the host-pathogen interactions during Escherichia coli mastitis, we first determined whether E. coli infection would change blood and milk polymorphonuclear neutrophil (PMN) chemiluminescence (CL) and viability. We then hypothesized that when E. coli invade the mammary gland, the viable PMN in milk would efficiently phagocytose and destroy E. coli before establishment of infection. We observed that the phagocytosis-dependent and independent CL were closely linked to PMN viability and were crucial to the outcome of mastitis. Maximal PMN influx and colony-forming units in infected quarters appeared at postinfection hours (PIH) 6 to 24. This further boosted PMN recruitment through bone marrow-blood barrier as well as blood-milk barrier. The survival of recruited PMN in the E. coli-infected quarters was much higher than that of noninfected quarters. Chemiluminescence activity of PMN from the infected quarters significantly increased following E. coli infection, even exceeding that of blood at PIH 6, 12, and 18 to 24; no such increase was observed in noninfected quarters, suggesting that the various responses of milk PMN to stimuli resulted largely from PMN viability. The highest CL intensity and durability was observed in milk PMN from infected quarters at PIH 12. Whereas an increased viability of PMN in the noninfected quarters was only significant at PIH 6, the viability of PMN in infected quarters was long lasting and significantly higher at PIH 6 to 72. Importantly, higher preinfection milk PMN viability correlated with bacterial clearance, which was accompanied by faster recovery. Our study strongly supports the hypothesis that boosting milk PMN viability could be a strategy with which to prevent or reduce the severity of coliform mastitis in dairy cows. This strategy might be achieved through strengthening bone marrow functionality. PMID- 15545378 TI - Effects of cleaning duration and water temperature on oxytocin release and milk removal in an automatic milking system. AB - Four different methods of teat preparation during milking in an automatic milking system were studied in 2 experiments on Red Holstein/German Fleckvieh cross-breed cows. Milking routines used were milking: 1) without premilking teat preparation; 2) with one cleaning cycle (58 to 60 s) with cold (13 to 15 degrees C) water; 3) with one cleaning cycle with warm water (30 to 32 degrees C); or 4) with 2 cleaning cycles (122 s) with warm water. In experiment 1, milking characteristics were evaluated and milking routines were randomly assigned to 62 cows during 3 measuring periods of 24 h each. In experiment 2, 10 randomly selected cows were assigned to the same milking routines during 4 d and blood samples for oxytocin (OT) determination were taken during milking in addition to milk flow recording. Milk production, peak flow rate, total, and quarter milk yields showed no differences among treatments. Premilking preparation with cold water compared with warm water showed no differences in OT release, milk yield, peak flow rate, main milking time, average flow rate, or time until main milk flow. Baseline OT concentrations were consistently low. At the start of teat cup attachment without premilking teat preparation OT concentrations remained on the basal level but were elevated in all other treatments. By 30 s from the start of milking, OT concentrations were markedly increased in all treatments and were no longer different between treatments. In conclusion, the teat cleaning device used in the automatic milking system, either with warm or cold water, was suitable to induce milk ejection in cows before the start of milking. PMID- 15545379 TI - Effects of mechanical processing on the nutritive value of barley silage for lactating dairy cows. AB - Mechanical processing of whole crop barley before ensiling may be useful for improving nutrient use by dairy cattle. The objective of this study was to assess the effects of feeding mechanically processed barley silage as the main forage source on lactational performance. Twenty-four Holstein cows, 16 primiparous (187 +/- 52 days in milk) and 8 multiparous (87 +/- 69 days in milk) cows, were used in a completely randomized design with a 2-wk covariate period and a 6-wk treatment period. The 2 treatments were: 1) total mixed ration (TMR) containing regular barley silage (RBS-TMR), and 2) TMR containing mechanically processed barley silage (MPBS-TMR). Barley silage and alfalfa hay supplied 41 and 5% of the dietary dry matter (DM), respectively. Intake, body weight, and milk production were measured during the covariate and treatment periods. In addition, 2 multiparous cows were used for in situ measurements of the ruminal DM and fiber degradation kinetics of the barley silages and TMR. Data were analyzed with repeated measurements using a mixed model that included the covariate adjustment. Feeding MPBS-TMR had no significant effects on DM intake (DMI; 21.7 kg/d), milk yield (33.9 kg/d), or milk composition, with only 4% FCM (fat-corrected milk) yield (29.7 vs. 31.7 kg/d) and milk fat concentration (3.30 vs. 3.57%) showing a numerical improvement. Apparent digestibilities of DM and nutrients were not affected by feeding MPBS-TMR, with the exception of starch digestibility, which tended to increase. Dairy efficiencies calculated as milk yield/DMI or FCM/DMI were not different between treatments. Body weight and body condition score were not affected by treatments. Effective ruminal degradability of DM was similar for both barley silages, indicating that when the silages were ground to remove the effects of mechanical processing, the potential digestion was similar. Mechanical processing of barley silage harvested at a mid-dough stage of maturity resulted in small improvements in its nutritive value for lactating dairy cows and had minor impact on digestibility and milk production. PMID- 15545380 TI - Feed efficiency of mid-lactation dairy cows fed yeast culture during summer. AB - Thirty-eight Holstein cows (26 multiparous and 12 primiparous), that averaged 105 d postpartum at the start of the experiment, were used to evaluate the feeding of yeast culture (60 g/cow daily of Diamond V XP) on production efficiency during hot summer weather. From early June until early September and after a 2-wk covariate period, cows were fed a control diet without or with 60 g of yeast culture/cow daily for 12 wk. Weekly daytime high temperatures in the free-stall barn during the 12-wk period averaged 33 degrees C (28 to 39 degrees C). Total mixed diets on a dry matter (DM) basis consisted of corn silage (28%), alfalfa hay (21%), and a concentrate mix (51%) without or with the yeast culture added to the total mixed ration at the time of feeding. Milk production (34.9 and 35.4 kg/d, for control and yeast culture treatment, respectively), 4% fat-corrected milk (31.2 and 32.0 kg/d), energy-corrected milk (ECM; 33.4 and 34.2 kg/d), and DM intake (23.1 and 22.1 kg/d) were similar for cows fed control and yeast culture diets. Percentages of milk fat (3.34 and 3.41) and true protein (2.85 and 2.87) were similar for both diets. Feed efficiency defined as kilogram of ECM/kilogram of DM intake was improved by 7% for cows fed the yeast culture. Body weights and body condition scores were similar for both groups. The results suggest that the yeast culture can improve feed efficiency of heat stressed dairy cows in midlactation. PMID- 15545381 TI - Effect of a monensin controlled-release capsule on rumen and blood metabolites in Florida Holstein transition cows. AB - The objective of this study was to determine the effect of a monensin controlled release capsule administered intraruminally at dry-off on rumen volatile fatty acids, NH3, lactate, pH, and energy blood metabolites in transition dairy cows fed Florida typical diets. In March 2003, 24 cows (10 primiparous and 14 multiparous) dried-off 50 to 70 d before expected parturition were randomly assigned to a treatment (n=12, oral capsule of monensin) or a control group (n=12, no capsule). Both groups received the same diet and were exposed to the same environment and management conditions. At assignment, at 21 d before expected parturition, at calving, and at 7, 14, and 21 d postpartum, blood samples were taken and body condition scores were determined. At 10 d postpartum, rumen and blood samples were obtained in the morning before the first feeding and at 2, 4, and 6 h after feeding. Serum nonesterified fatty acids (NEFA), beta hydroxybutyrate (BHBA), and glucose were measured. Rumen samples were analyzed for concentrations of acetic, propionic, butyric, L- and D-lactic acids, and NH3. Data for rumen and blood metabolites were analyzed by ANOVA, mixed models for repeated measures. Volatile fatty acids were not different between groups. Multiparous treated cows had a significant reduction in rumen NH3 at 6 h after feeding. Treatment with monensin significantly increased body condition score at calving in multiparous cows. During the postpartum period, NEFA and BHBA were noticeably lower in treated than in control primiparous cows. This difference was not observed in multiparous cows. PMID- 15545382 TI - Competition for teats and feeding behavior by group-housed dairy calves. AB - Many farms using teat-based systems for supplying milk for calves provide only one or a small number of teats for a group of calves, but no previous research has addressed how competition for teats affects calf behavior or milk intake. The aim of this study was to determine how restricted access to teats affects calf competitive behavior, meal-based feeding patterns, and milk intake. Female calves (n=15) were divided into 5 groups of 3 calves each and fed with a teat-to-calf ratio that varied daily from 1:3 to 4:3 using a switchback design. Feeding behavior was recorded by scoring the time and duration of each sucking event. We defined meals using the frequency distribution of log intervals between visits to the teat and identified the within-meal and between-meal distributions intersection points. Three classes of intervals were identified based on the intersection points of the distributions: 1) intervals <2 min, representing small breaks away from the teat within a meal; 2) intervals >41 min, providing an objective definition of a new meal; and 3) an intermediate distribution of intervals from 2 to 41 min could be included in either of the other 2 classes. Meal number showed no significant decrease with decreasing teat number. However, total time on the teat decreased from 40.2 to 32.7 (+/-2.6) min/d, and milk consumption declined from 14.0 to 11.4 (+/-0.8) L/d as teat number declined from 4 to 1. In addition, competitive interactions became more frequent when teat access was reduced; the number of times calves displaced one another from a teat increased from 18 to 41 (+/-5) times/d when teat number decreased from 4 to 1. In conclusion, reduced access to teats increases competitive interactions, decreases feeding time and decreases milk intake by group-housed calves. PMID- 15545383 TI - Feeding glycerol to transition dairy cows: effects on blood metabolites and lactation performance. AB - Glycerol can alleviate the symptoms of ketosis when delivered as an oral drench. The addition of glycerol to the diet would eliminate the need for restraining cows for drenching yet deliver a glucogenic substrate, alleviate the fatty liver ketosis complex, and improve lactational performance. For this study, 21 multiparous and 9 primiparous Holstein cows blocked by parity and expected calving date were used in a randomized block design to evaluate the effects of feeding glycerol from 14 d prepartum to 21 d in milk (DIM). Treatments (kg/d dry matter basis) were 0.86 of cornstarch (control), 0.43 cornstarch + 0.43 glycerol (LG), or 0.86 glycerol (HG), topdressed and hand-mixed into the upper one-third of the daily ration. All cows were fed a common diet from 22 to 70 DIM. Prepartum dry matter intake (DMI) was greater for cows fed the control diet compared with LG or HG (13.3, 10.8, and 11.3 +/- 0.50 kg/d, respectively). Prepartum plasma glucose, insulin, beta-hydroxybutyrate, nonesterified fatty acids, and ruminal profiles were not affected by treatments. Rumen fluid collected postpartum from cows fed LG and HG had greater total volatile fatty acids, greater molar proportions of propionate, and a decreased ratio of acetate to propionate. Furthermore, concentrations of butyrate tended to be greater in rumens of cows fed LG and HG. Postpartum concentrations of glucose in plasma were greatest for cows fed the control diet relative to LG and HG (66.0 vs. 63.1 and 58.4 mg/dL, respectively) and decreased sharply at 21 DIM, after treatments ended, for cows fed HG (diet x day interaction). Body weight and condition loss, plasma nonesterified fatty acids, and liver lipids during the first 21 DIM were similar among treatments. Postpartum DMI was not affected by treatments; however, a tendency was observed for a diet x day interaction for body weight, as cows fed LG gained more body weight from 21 to 70 DIM relative to cows fed HG. Yield of energy-corrected milk during the first 70 DIM tended to be greatest for cows fed the control diet. The LG and HG diets decreased urea nitrogen concentrations in milk relative to controls. Based upon prepartum DMI and concentrations of glucose and beta-hydroxybutyrate in blood postpartum, feeding glycerol to dairy cows at the levels used in this experiment increased indicators used to gauge the degree of ketosis in dairy cattle. PMID- 15545384 TI - Technical note: a computerized system for monitoring feeding behavior and individual feed intake of dairy cattle. AB - The objective of this study was to develop and validate a computerized system to monitor feeding behavior and feed intake of loose-housed dairy cattle. The system consisted of 28 scales located in front of each self-locking place of a regular feedbunk. All cows had access to all scales indifferently. Each visit to the feedbunk was monitored by a transponder in the ear of each cow that was detected by a proximity reader located at the top right corner of each headlock. The data from the scales and the proximity readers were continuously recorded by a computer with an average scanning time of 3.5 s. The monitoring system was validated using all 28 feeding places and 51 lactating cows in a series of 4-h observations during 5 different d. During the observation periods, for each feeder, 2 observers recorded the cow number and the exact time of the visit. The observed data were then compared with the computer records. To validate the ability of the system to monitor feed consumption, on separate days, the amount of feed consumed by a cow during a visit was also measured manually with an external scale, and the feed that disappeared from each scale in 2 different 24-h periods was compared with the sum of feed consumed in each scale during these 2 periods. The average time spent in a given scale by each cow determined by direct observations was similar to that determined by the computer. The system was accurate and showed a high specificity (98.8%) and sensitivity (99.6%) for cow detections. Feed weights determined by the computer system were similar to those measured manually with an external scale, implying that the system was also accurate in measuring individual intake weights. In conclusion, the system provided a reasonable estimate of the number of visits per animal, length of each visit, amount of feed consumed per visit and animal, the total amount of feed consumed daily by each animal, and the rate at which animals consume feed. PMID- 15545385 TI - Peripartal metabolism and production of holstein cows fed diets supplemented with fat during the dry period. AB - Previous research from our laboratory demonstrated that cows fed supplemental fat throughout the dry period in an attempt to increase body condition score (BCS) had little hepatic lipid accumulation at d 1 postpartum compared with cows fed an isocaloric high-grain diet or a lower energy control diet. However, results were confounded by lower dry matter intake and loss of BCS by cows fed the fat supplemented diet. Here, cows were fed a control diet (C) moderately high in nonfiber carbohydrates (NFC) or an isocaloric fat-supplemented, low NFC (F) diet to reassess the effects of supplemental fat throughout the dry period on peripartal lipid accumulation in liver. A more energy-dense, high-NFC diet supplemented with fat (CF) was also fed to test the efficacy of supplemental fat in a diet with similar carbohydrate composition but higher energy density. Intakes of dry matter and net energy for lactation were similar among treatments throughout the experiment, although diet x day interactions during the last 21 d before parturition indicated that cows fed CF decreased intakes more slowly. Cows gained similar amounts of BCS and body weight among diets prepartum, but cows fed C tended to lose more BCS and body weight around parturition. Milk production and milk components did not differ among treatments. Prepartum concentrations of glucose, insulin, total protein, nonesterified fatty acids, and mu hydroxybutyrate in plasma were similar among treatments. Supplemental fat increased prepartum concentrations of urea and cholesterol in plasma. Postpartum concentrations of metabolites and insulin in plasma were similar among treatments. Concentrations of total lipid and triglyceride in liver increased at parturition, whereas hepatic glycogen concentration decreased, but concentrations were not different among treatments. Supplemental fat fed prepartum did not affect peripartal lipid accumulation in liver tissue and did not benefit postpartum milk production. PMID- 15545386 TI - Effects of replacing dietary starch with sucrose on ruminal fermentation and nitrogen metabolism in continuous culture. AB - A dual-effluent continuous-culture system was used to evaluate the effects of partially replacing cornstarch with sucrose in a total mixed ration on ruminal fermentation and N metabolism. The 4 treatments were 0 (control), 2.5, 5.0, and 7.5% sucrose and, respectively, 7.5 (control), 5.0, 2.5, and 0% cornstarch in a total mixed ration containing 20% corn silage and 40% alfalfa silage. Fermenters were fed 4 times a day during four 9-d periods with sampling beginning on d 6. Replacing cornstarch with sucrose did not alter ruminal pH (5.97), total volatile fatty acids (VFA) (104.4 mmol/L), or the acetate to propionate ratio (2.16); however, branched-chain volatile fatty acids were higher for the control treatment compared with the 7.5% sucrose treatment. Five hours postfeeding, sucrose treatments significantly altered molar proportions of all volatile fatty acids, and acetate-to-propionate and glucogenic-to-lipogenic ratios. Digestibility of dry matter and N were not affected by treatment, but digestibility of total non-structural carbohydrates was increased with sucrose treatments. A quadratic effect was noted for neutral detergent fiber (NDF) digestibility as sucrose replaced starch. A higher NDF digestibility (66.1 vs. 59.9%) was observed for the 7.5% sucrose treatment compared with the other 2 sucrose treatments. Levels of ammonia N were within an acceptable range to support microbial protein synthesis and did not differ among treatments (mean=9.23 mg/dL). Sucrose inclusion in the total mixed ration did not affect bacterial N synthesis. Results indicate that (at the levels tested in this study) inclusion of sucrose in the diet when rumen-degradable protein is adequate does not affect ruminal fermentation. PMID- 15545387 TI - Real-time PCR quantification of bovine lactase mRNA: localization in the gastrointestinal tract of milk-fed calves. AB - Lactase is a disaccharidase that is present in the brush-border membrane of the small intestine, hydrolyzes lactose to glucose and galactose, and is therefore important in milk-fed animals. Assays based on quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) in the bovine species have not yet been described. Therefore, we have developed an RT-PCR assay for the quantification of lactase mRNA levels and have tested its suitability in the bovine gastrointestinal tract of seven 5-d-old milk-fed calves. Primers for RT PCR amplification of bovine lactase mRNA were designed in the 100% identical regions of species (rats, rabbits, humans) from which lactase sequences were available. Lactase mRNA was expressed relative to mean levels of 4 housekeeping genes (glyceraldehyde-3-phosphate dehydrogenase, beta-actin, ubiquitin, and 18S). The presence of lactase mRNA along the entire gastrointestinal tract was evaluated in samples that consisted of whole gut walls (mucosa plus submucosa). Furthermore, mRNA levels of lactase were measured in fractionized layers of jejunal and ileal mucosa (mainly containing villus tips or crypts) and ileal lamina propria (mainly containing Peyer's patches). Agarose gel electrophoresis of the lactase PCR product revealed a single band that corresponded to the single amplified product as predicted by the melting curve analysis of the PCR. The amplified partial-bovine lactase sequence showed 87% similarity with human and rabbit sequences and 82% similarity with the rat sequence. Lactase mRNA was present in whole walls (consisting of mucosa and submucosa) of the entire small intestine, but was absent in esophagus, rumen, fundus, pylorus, and colon. Furthermore, lactase mRNA was detected in fractionized villus and crypt layers of jejunum and ileum, but levels were higher in the jejunum in villus than in crypt fractions. No lactase mRNA was detectable in the lamina propria fraction of the ileum containing mainly Peyer's patches. In conclusion, the developed RT-PCR method allows study of lactase mRNA levels. PMID- 15545388 TI - Fermentation characteristics of corn forage ensiled in mini-silos. AB - To evaluate numerous experimental variables and their interactions involving different corn (Zea mays, L.) silage hybrids, scaled down mini-silos are necessary. Objectives of this study were to evaluate the influence of sample size on pH, NH3, and volatile fatty acid profile of 8 corn silage hybrids, selected to vary in fiber digestibility and ensiled in vacuum-sealed polyethylene bags for 90 d, and to assess the suitability of these mini-silos for detecting differences among corn silage samples. Hybrids were grown at the Cornell Teaching and Research Center located near Harford, NY, and harvested at a dry matter content of about 32% in the fall of 2002. Samples from 3 field replications of each hybrid were chipper-shredder chopped and vacuum-ensiled in bags with sample sizes of 50, 100, 200, 400, and 600 g. Increasing sample size resulted in decreased lactic acid, acetic acid, total acids, and NH3. Most of the difference among sample sizes occurred between the 50- and 100-g sample sizes. Lactic acid:acetic acid ratio (3.1 +/- 0.13) and pH (3.9 +/-0.08) did not vary among sample sizes. There was no detectable butyric acid in the samples. Fermentation characteristics suggested that all samples were well ensiled but that the fermentation profile of the 50-g samples differed the most from other sample sizes. Hybrids did vary in lactic acid, acetic acid, lactic acid:acetic acid, and pH. Differences among hybrids were also noted for dry matter and crude protein. Field-chopped corn hybrids that were ensiled using mini-silos had higher acids than corresponding field-chopped corn hybrids ensiled in Ag-bags, in part due to no effluent escaping from the mini-silos. It is possible to use vacuum-sealed plastic bags to ensile corn, with samples as small as 200 g, and to use these mini-silos to assess differences among corn silage samples. Caution should be used when extrapolating mini-silo data to field-scale ensiling. PMID- 15545389 TI - A model of net amino acid absorption and utilization by the portal-drained viscera of the lactating dairy cow. AB - A more complete understanding of amino acid (AA) metabolism by the various tissues of the body is required to improve upon current systems for predicting the use of absorbed AA. The objective of this work was to construct and parameterize a model of net removal of AA by the portal-drained viscera (PDV). Six cows were prepared with arterial, portal, and hepatic catheters and infused abomasally with 0, 200, 400, or 600 g of casein daily. Casein infusion increased milk yield quadratically and tended to increase milk protein yield quadratically. Arterial concentrations of a number of essential AA increased linearly with respect to infusion amount. When infused casein was assumed to have a true digestion coefficient of 0.95, the minimum likely true digestion coefficient for noninfused duodenal protein was found to be 0.80. Net PDV use of AA appeared to be linearly related to total supply (arterial plus absorption), and extraction percentages ranged from 0.5 to 7.25% for essential AA. Prediction errors for portal vein AA concentrations ranged from 4 to 9% of the observed mean concentrations. Removal of AA by PDV represented approximately 33% of total postabsorptive catabolic use, including use during absorption but excluding use for milk protein synthesis, and was apparently adequate to support endogenous N losses in feces of 18.4 g/d. As 69% of this use was from arterial blood, increased PDV catabolism of AA in part represents increased absorption of AA in excess of amounts required by other body tissues. Based on the present model, increased anabolic use of AA in the mammary and other tissues would reduce the catabolic use of AA by the PDV. PMID- 15545390 TI - Supplementing intensively grazed late-gestation and early-lactation dairy cattle with chromium. AB - Two hundred thirty-two primiparous and multiparous cows were assigned to a study to determine the effect of supplementing 0 or 6.25 mg/d of Cr from Cr Met on lactation and reproductive performance. Cows received treatments from 6 wk precalving through 21 wk postpartum. Precalving, treatments were incorporated into a pelleted grain mixture and group-fed. Post-calving, cows received treatments via an individual oral drench once a day after the a.m. milking. Grazed herbage was the primary diet constituent for lactating cattle. Blood was collected from a predetermined group of cows before and immediately after calving. On 4 occasions during the treatment period, milk yield was recorded and samples collected for determination of composition. Chromium supplementation had no effect on yield of milk and milk components and milk composition. Chromium supplementation decreased serum nonesterified fatty acids (NEFA) concentration (0.60 vs. 0.68 mmol/L), with chromium supplementation having the greatest impact on serum NEFA concentrations at 1 wk prepartum. Greater percentages of cows supplemented with Cr were observed to be anestrus by dairy personnel (45.5 vs. 32.0%). However, Cr supplementation tended to increase the percentage of cows pregnant in the first 28 d of the mating season (50.0 vs. 39.2%). Results indicate that Cr Met supplementation of intensely grazed, late-gestation and early-lactation dairy cattle decreased serum NEFA concentrations and tended to increase pregnancy rates in the first 28 d of the mating season. PMID- 15545391 TI - Effects of dry matter intake, addition of buffer, and source of fat on duodenal flow and concentration of conjugated linoleic acid and trans-11 C18:1 in milk. AB - The primary objective of the study was to investigate the effects of DM intake, addition of buffer, and fish vs. soybean oil on duodenal flows and milk concentrations of conjugated linoleic acid (CLA) and trans-11 C18:1. Four ruminally and duodenally cannulated multiparous cows averaging 106 +/- 17 d in milk at the start of the trial were used in a 4 x 4 Latin square design with treatments as follows: 1) control=diet contained 2% fish oil and fed ad libitum, 2) buffer addition (BUFF)=control diet with 0.8% of NaHCO3 added, 3) low DM intake (LDMI)=DMI restricted to 80% of the control but concentration of fish oil was increased to 2.5% to provide for similar fatty acids (FA) intake, and 4) soybean oil (SBO)=same as control except 2% soybean oil was substituted for fish oil. The diets consisted of 36.2% forage and 63.8% concentrate. Each period consisted of 18 d, with the last 7 d devoted to data collection and the first 4 d used to determine the appropriate amount of feed to be offered to the cow on LDMI. Duodenal flows of CLA and trans-C18:1 were lower for SBO than for diets with fish oil. Feeding buffer did not affect ruminal pH or duodenal flows of trans-11 C18:1 and CLA. Restriction of DMI decreased duodenal flow of trans-11 C18:1 but did not decrease duodenal flow of CLA compared with control. In milk, CLA concentration was lower for SBO (24.5, 17.9, 18.5, and 10.1 mg/g of FA for control, BUFF, LDMI, and SBO, respectively). Cows fed fish oil had higher duodenal flow and milk concentration of n-3 polyunsaturated fatty acids than the cows fed SBO. Compared with SBO, fish oil is more effective in increasing duodenal flows of CLA and trans-11 C18:1, and thus, concentration of CLA in milk. PMID- 15545392 TI - Genetic selection for health traits using producer-recorded data. I. Incidence rates, heritability estimates, and sire breeding values. AB - The objective of this study was to determine the feasibility of genetic selection for health traits in dairy cattle using data recorded in on-farm herd management software programs. Data regarding displaced abomasum (DA), ketosis (KET), mastitis (MAST), lameness (LAME), cystic ovaries (CYST), and metritis (MET) were collected between January 1, 2001 and December 31, 2003 in herds using Dairy Comp 305, DHI-Plus, or PCDART herd management software programs. All herds in this study were either participants in the Alta Genetics (Watertown, WI) Advantage progeny testing program or customers of the Dairy Records Management Systems (Raleigh, NC) processing center. Minimum lactation incidence rates were applied to ensure adequate reporting of these disorders within individual herds. After editing, DA, KET, MAST, LAME, CYST, and MET data from 75,252 (313), 52,898 (250), 105,029 (429), 50,611 (212), 65,080 (340), and 97,318 (418) cows (herds) remained for analysis. Average lactation incidence rates were 0.03, 0.10, 0.20, 0.10, 0.08, and 0.21 for DA, KET, MAST, LAME, CYST, and MET (including retained placenta), respectively. Data for each disorder were analyzed separately using a threshold sire model that included a fixed parity effect and random sire and herd year-season of calving effects; both first lactation and all lactation analyses were carried out. Heritability estimates from first lactation (all lactation) analyses were 0.18 (0.15) for DA, 0.11 (0.06) for KET, 0.10 (0.09) for MAST, 0.07 (0.06) for LAME, 0.08 (0.05) for CYST, and 0.08 (0.07) for MET. Corresponding heritability estimates for the pooled incidence rate of all diseases between calving and 50 d postpartum were 0.12 and 0.10 for the first and all lactation analyses, respectively. Mean differences in PTA for probability of disease between the 10 best and 10 worst sires were 0.034 for DA, 0.069 for KET, 0.130 for MAST, 0.054 for LAME, 0.039 for CYST, and 0.120 for MET. Based on the results of this study, it appears that genetic selection against common health disorders using data from on-farm recording systems is possible. PMID- 15545394 TI - A Monte Carlo approach for estimation of haplotype probabilities in half-sib families. AB - The objective of this work was to propose an algorithm (HAPROB) to estimate haplotype probabilities for genotyped members of half-sib families for which parents lacked genotypic information. The algorithm had 2 basic steps. First, a Monte Carlo-based approach was used to estimate haplotype probabilities for sires conditional upon offspring genotypes and population allelic frequencies, and then offspring-haplotype probabilities were estimated conditional upon sire probabilities and population frequencies. The 2 steps were alternated iteratively until estimates of population frequencies were essentially unchanged. Simulation was used to evaluate effects of the number of Monte Carlo cycles on the accuracy of the reconstructed haplotypes. Fifty thousand cycles was found to be sufficient for the haplotype configurations considered. Accuracy of the algorithm was compared with that obtained by the public domain SIMWALK2 software. Predictions of the most likely haplotype configurations are produced by SIM-WALK2, but no estimates of probability are given. The accuracy of the current approach was comparable to that obtained from SIMWALK2. The proportions of times that haplotypes were reconstructed correctly were 87.0 and 92.4% (sires and offspring) for HAPROB vs. 87.5 and 91.5% for SIMWALK2. Effects of family size on accuracy of reconstruction were examined. Accuracy of reconstruction was only about 4% for sires with 2 offspring, but accuracy among the offspring themselves was 65%. Accuracy increased quickly as family size increased and reached 100% for sires with 30 offspring. Maximum accuracy for offspring was about 96%. Estimates of haplotype probabilities produced can be used in regression analyses to estimate effects of haplotypes on quantitative phenotypes. PMID- 15545393 TI - Genetic selection for health traits using producer-recorded data. II. Genetic correlations, disease probabilities, and relationships with existing traits. AB - The objectives of this study were to calculate genetic correlations between health traits that were recorded in on-farm herd management software programs and to assess relationships between these traits and other traits that are routinely evaluated in US dairy sires. Data consisted of 272,576 lactation incidence records for displaced abomasum (DA), ketosis (KET), mastitis (MAST), lameness (LAME), cystic ovaries (CYST), and metritis (MET) from 161,622 cows in 646 herds. These data were collected between January 1, 2001 and December 31, 2003 in herds using the Dairy Comp 305, DHI-Plus, or PCDART herd management software programs. Binary incidence data for all disorders were analyzed simultaneously using a multiple-trait threshold sire model that included random sire and herd-year season of calving effects. Although data from multiple lactations were available for some animals, our genetic analysis included only first parity records due to concerns about selection bias and improper modeling of the covariance structure. Heritability estimates for the presence or absence of each disorder during first lactation were 0.14 for DA, 0.06 for KET, 0.09 for MAST, 0.03 for LAME, 0.04 for CYST, and 0.06 for MET. Estimated genetic correlations were 0.45 between DA and KET, 0.42 between KET and CYST, 0.20 between MAST and LAME, 0.19 between KET and LAME, 0.17 between DA and CYST, 0.17 between KET and LAME, 0.17 between KET and MET, and 0.16 between LAME and CYST. All other correlations were negligible. Correlations between predicted transmitting abilities for the aforementioned health traits and existing production, type, and fitness traits were low, though it must be noted that these estimates may have been biased by low reliability of the health trait evaluations. Based on results of this study, it appears that genetic selection for health disorders recorded in on-farm software programs can be effective. These traits can be incorporated into selection indices directly, or they can be combined into composite traits, such as "reproductive disorders", "metabolic disorders", or "early lactation disorders". PMID- 15545395 TI - Effects of casein haplotypes on milk production traits in Italian Holstein and Brown Swiss cattle. AB - The objective of this study was to estimate the effects of different haplotypes of the casein genes on milk production traits in Italian dairy cattle. Traits of interest were yields of milk, fat, and protein, and percentages of fat and protein in milk. The data included 728 multiparous records from 347 Holsteins and 773 records from 298 Brown Swiss cows. Records were preadjusted for effects of age and parity, season of calving, and region, and expressed as deviations from herdmate averages. Twenty half-sib families were represented in each breed. Haplotype probabilities were estimated for each animal and phenotypes were regressed on these probabilities. Nine haplotypes were observed in Holsteins and 17 were identified among the Brown Swiss. For Holsteins, significant effects were observed for protein percentage, with some indication of an effect for fat percentage. For the Brown Swiss, effects of haplotypes were significant for milk yield and fat and protein percentages. Effects were strongest for protein percentage. Correlation coefficients of solutions across breeds tended to be strong and positive, indicating that the same haplotypes had similar estimated effects in the 2 breeds. Although the data were limited (<350 cows in each study), this latter result may suggest that genes in the casein complex itself are responsible for the effects observed, rather than loci that are physically linked on either side of the casein cluster. PMID- 15545396 TI - Genotype and diet effects on energy balance in the first three lactations of dairy cows. AB - Dairy cows make strategic use of body energy to support early lactation and replenish this lost energy later in lactation, thereby creating body energy profiles that vary both within lactation and across lactations. The interaction between genotype and diet energy content is interesting from a management viewpoint and from a cow survival viewpoint. In this study, we modeled energy balance over 3 lactations using a multivariate random regression model, for cows from the Langhill Dairy Cattle Research Centre. This herd has been selected for maximum production or to remain at the UK national average for production (control group) and has been fed 2 diets of different energy density (high or low concentrate). Cows in the various groups differed in body condition score and the way they lost and regained body lipid. Cows selected for maximum production on a low-concentrate diet lost the greatest amount of body lipid (0.54 condition score units) after 3 lactations. Cows selected for maximum production lost more than control cows on either diet, indicating that selection mainly for yield has predisposed cows to utilize body energy to support lactation on diets spanning the range used here. Cows selected for maximum production were heavier at first calving than control cows but lost more weight and regained more weight so that at the end of each lactation, cows in the 2 groups did not differ significantly in weight. Cows use body lipid to support lactation over 3 lactations and the profile of that use varies according to genotype and diet. PMID- 15545397 TI - Genetic parameters for days open and pregnancy rates in US Holsteins using different editing criteria. AB - The influence of various editing criteria for days open (DO) records on genetic parameter estimates of DO and pregnancy rates (PR) in US Holsteins was investigated. Data included first parity 305-d milk yield and DO records from 8 states: Georgia (GA), Florida (FL), North Carolina (NC), Texas (TX), Arizona (AZ), California (CA), New York (NY), and Wisconsin (WI). The pregnancy rate was computed as 1/[(DO - VWP)/HI + 1)], where VWP was the approximate voluntary waiting period and HI was the heat interval set as 21 d. The upper limit for PR was set to 1.0. A bivariate animal model for DO (or PR) and 305-d milk yield was fit separately for each state. The model included fixed effects of herd-year, month of calving, and age of cow, as well as random animal and residual effects. In separate analyses, maximum DO records were limited to 150, 200, 250, 300, and 365 d. Analyses for PR used values of 50, 80, and 120 d for the VWP. Genetic and residual variances for DO were strongly dependent on the upper limit; both variances were 8 times larger as the upper bound increased from 150 to 365 d. Estimates of heritability for DO varied between 0.03 and 0.06. There was a 30% increase in the heritability estimate as the upper limit increased from 150 to 250 d for FL and NC, and small or no increases for the other states. The increase of the upper limit from 250 to 365 d resulted in little change. The genetic correlation between milk and DO was the highest for FL (0.6) and the lowest for GA (0.12 to 0.23). For PR with VWP=50, the heritability was higher than the corresponding estimate for DO in GA, equal to that in AZ, and lower in the remaining states. Heritabilities of PR also varied by the length of VWP; highest heritabilities were obtained at VWP=50 d for GA and AZ; at VWP=80 d for NY and WI; at VWP=120 d for FL, NC, and CA. Increase of genetic variation for records of DO < 250 d was small. Days open and PR are strongly influenced by differences in management protocols among states. PMID- 15545398 TI - Phosphorus concentration and solubility in dairy feces: variability and affecting factors. AB - Recent data from phosphorus (P) feeding trials have demonstrated that P concentration in dairy feces is directly affected by P levels in diets and that farm P surpluses as well as potential environmental losses can be reduced through dietary manipulation. The current study was conducted to examine the variability of fecal P under farm conditions and to elucidate factors affecting the concentration and solubility of fecal P. Feed and fecal samples from >30 commercial dairies in the Northeast and Mid-Atlantic regions were analyzed. Dietary P concentrations ranged from 3.45 to 5.78 g/kg of feed DM (DM), and P determined in acid digests (TP) of feces from 5.84 to 12.84 g/kg of fecal DM. On average, 50% of fecal TP was water soluble; of the latter, 83% was inorganic (Pi). Across-farm variability (n=33) had CV averaging 18.9% for fecal TP and >20% for Pi and total P (Pt) in water extracts. Within-farm variability based on multiple samples per herd had the same magnitude as across-farm and was independent of sample numbers from individual farms (n=7 to 30). Of all fecal parameters determined, pH and DM had the lowest variability (CV <10%), water soluble Pi, Pt, and Ca the highest (CV of 20 to 30%), and total P, Ca, and Mg determined by acid digests were intermediate (CV 10 to 20%). Water-soluble Pi concentrations determined in dried-ground fecal samples were lower than in wet samples. The drying-grinding process changes Pi solubility and the change is not linear. This study confirms that dietary P concentration is the dominating factor affecting fecal P excretion; however, Ca concentration, DIM, and fecal pH also made small, but statistically significant contributions, although some of the mechanisms remain to be thoroughly investigated. PMID- 15545399 TI - Luminal Ca2+-regulated Mg2+ inhibition of skeletal RyRs reconstituted as isolated channels or coupled clusters. AB - In resting muscle, cytoplasmic Mg(2+) is a potent inhibitor of Ca(2+) release from the sarcoplasmic reticulum (SR). It is thought to inhibit calcium release channels (RyRs) by binding both to low affinity, low specificity sites (I-sites) and to high affinity Ca(2+) sites (A-sites) thus preventing Ca(2+) activation. We investigate the effects of luminal and cytoplasmic Ca(2+) on Mg(2+) inhibition at the A-sites of skeletal RyRs (RyR1) in lipid bilayers, in the presence of ATP or modified by ryanodine or DIDS. Mg(2+) inhibits RyRs at the A-site in the absence of Ca(2+), indicating that Mg(2+) is an antagonist and does not simply prevent Ca(2+) activation. Cytoplasmic Ca(2+) and Cs(+) decreased Mg(2+) affinity by a competitive mechanism. We describe a novel mechanism for luminal Ca(2+) regulation of Ca(2+) release whereby increasing luminal [Ca(2+)] decreases the A site affinity for cytoplasmic Mg(2+) by a noncompetitive, allosteric mechanism that is independent of Ca(2+) flow. Ryanodine increases the Ca(2+) sensitivity of the A-sites by 10-fold, which is insufficient to explain the level of activation seen in ryanodine-modified RyRs at nM Ca(2+), indicating that ryanodine activates independently of Ca(2+). We describe a model for ion binding at the A-sites that predicts that modulation of Mg(2+) inhibition by luminal Ca(2+) is a significant regulator of Ca(2+) release from the SR. We detected coupled gating of RyRs due to luminal Ca(2+) permeating one channel and activating neighboring channels. This indicated that the RyRs existed in stable close-packed rafts within the bilayer. We found that luminal Ca(2+) and cytoplasmic Mg(2+) did not compete at the A-sites of single open RyRs but did compete during multiple channel openings in rafts. Also, luminal Ca(2+) was a stronger activator of multiple openings than single openings. Thus it appears that RyRs are effectively "immune" to Ca(2+) emanating from their own pore but sensitive to Ca(2+) from neighboring channels. PMID- 15545400 TI - Gating charges in the activation and inactivation processes of the HERG channel. AB - The hERG channel has a relatively slow activation process but an extremely fast and voltage-sensitive inactivation process. Direct measurement of hERG's gating current (Piper, D.R., A. Varghese, M.C. Sanguinetti, and M. Tristani-Firouzi. 2003. PNAS. 100:10534-10539) reveals two kinetic components of gating charge transfer that may originate from two channel domains. This study is designed to address three questions: (1) which of the six positive charges in hERG's major voltage sensor, S4, are responsible for gating charge transfer during activation, (2) whether a negative charge in the cytoplasmic half of S2 (D466) also contributes to gating charge transfer, and (3) whether S4 serves as the sole voltage sensor for hERG inactivation. We individually mutate S4's positive charges and D466 to cysteine, and examine (a) effects of mutations on the number of equivalent gating charges transferred during activation (z(a)) and inactivation (z(i)), and (b) sidedness and state dependence of accessibility of introduced cysteine side chains to a membrane-impermeable thiol-modifying reagent (MTSET). Neutralizing the outer three positive charges in S4 and D466 in S2 reduces z(a), and cysteine side chains introduced into these positions experience state-dependent changes in MTSET accessibility. On the other hand, neutralizing the inner three positive charges in S4 does not affect z(a). None of the charge mutations affect z(i). We propose that the scheme of gating charge transfer during hERG's activation process is similar to that described for the Shaker channel, although hERG has less gating charge in its S4 than in Shaker. Furthermore, channel domain other than S4 contributes to gating charge involved in hERG's inactivation process. PMID- 15545401 TI - Block of inactivation-deficient Na+ channels by local anesthetics in stably transfected mammalian cells: evidence for drug binding along the activation pathway. AB - According to the classic modulated receptor hypothesis, local anesthetics (LAs) such as benzocaine and lidocaine bind preferentially to fast-inactivated Na(+) channels with higher affinities. However, an alternative view suggests that activation of Na(+) channels plays a crucial role in promoting high-affinity LA binding and that fast inactivation per se is not a prerequisite for LA preferential binding. We investigated the role of activation in LA action in inactivation-deficient rat muscle Na(+) channels (rNav1.4-L435W/L437C/A438W) expressed in stably transfected Hek293 cells. The 50% inhibitory concentrations (IC(50)) for the open-channel block at +30 mV by lidocaine and benzocaine were 20.9 +/- 3.3 microM (n = 5) and 81.7 +/- 10.6 microM (n = 5), respectively; both were comparable to inactivated-channel affinities. In comparison, IC(50) values for resting-channel block at -140 mV were >12-fold higher than those for open channel block. With 300 microM benzocaine, rapid time-dependent block (tau approximately 0.8 ms) of inactivation-deficient Na(+) currents occurred at +30 mV, but such a rapid time-dependent block was not evident at -30 mV. The peak current at -30 mV, however, was reduced more severely than that at +30 mV. This phenomenon suggested that the LA block of intermediate closed states took place notably when channel activation was slow. Such closed-channel block also readily accounted for the LA-induced hyperpolarizing shift in the conventional steady state inactivation measurement. Our data together illustrate that the Na(+) channel activation pathway, including most, if not all, transient intermediate closed states and the final open state, promotes high-affinity LA binding. PMID- 15545402 TI - Arachidonic acid inhibits epithelial Na channel via cytochrome P450 (CYP) epoxygenase-dependent metabolic pathways. AB - We used the patch-clamp technique to study the effect of arachidonic acid (AA) on epithelial Na channels (ENaC) in the rat cortical collecting duct (CCD). Application of 10 microM AA decreased the ENaC activity defined by NPo from 1.0 to 0.1. The dose-response curve of the AA effect on ENaC shows that 2 microM AA inhibited the ENaC activity by 50%. The effect of AA on ENaC is specific because neither 5,8,11,14-eicosatetraynoic acid (ETYA), a nonmetabolized analogue of AA, nor 11,14,17-eicosatrienoic acid mimicked the inhibitory effect of AA on ENaC. Moreover, inhibition of either cyclooxygenase (COX) with indomethacin or cytochrome P450 (CYP) omega-hydroxylation with N-methylsulfonyl-12,12 dibromododec-11-enamide (DDMS) failed to abolish the effect of AA on ENaC. In contrast, the inhibitory effect of AA on ENaC was absent in the presence of N methylsulfonyl-6-(propargyloxyphenyl)hexanamide (MS-PPOH), an agent that inhibits CYP-epoxygenase activity. The notion that the inhibitory effect of AA is mediated by CYP-epoxygenase-dependent metabolites is also supported by the observation that application of 200 nM 11,12-epoxyeicosatrienoic acid (EET) inhibited ENaC in the CCD. In contrast, addition of 5,6-, 8,9-, or 14,15-EET failed to decrease ENaC activity. Also, application of 11,12-EET can still reduce ENaC activity in the presence of MS-PPOH, suggesting that 11,12-EET is a mediator for the AA induced inhibition of ENaC. Furthermore, gas chromatography mass spectrometry analysis detected the presence of 11,12-EET in the CCD and CYP2C23 is expressed in the principal cells of the CCD. We conclude that AA inhibits ENaC activity in the CCD and that the effect of AA is mediated by a CYP-epoxygenase-dependent metabolite, 11,12-EET. PMID- 15545403 TI - Nonsteroidal selective androgen receptors modulators (SARMs): designer androgens with flexible structures provide clinical promise. PMID- 15545404 TI - Endogenous cannabinoids take the edge off neuroendocrine responses to stress. PMID- 15545405 TI - Endocrine-related resources from the National Institutes of Health. PMID- 15545406 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. AB - The purpose of this announcement is to effect the valid publication of the following new names and new combinations under the procedure described in the Bacteriological Code (1990 Revision). Authors and other individuals wishing to have new names and/or combinations included in future lists should send three copies of the pertinent reprint or photocopies thereof to the IJSEM Editorial Office for confirmation that all of the other requirements for valid publication have been met. It is also a requirement of IJSEM and the ICSP that authors of new species, new subspecies and new combinations provide evidence that types are deposited in two recognized culture collections in two different countries (i.e. documents certifying deposition and availability of type strains). It should be noted that the date of valid publication of these new names and combinations is the date of publication of this list, not the date of the original publication of the names and combinations. The authors of the new names and combinations are as given below, and these authors' names will be included in the author index of the present issue and in the volume author index. Inclusion of a name on these lists validates the publication of the name and thereby makes it available in bacteriological nomenclature. The inclusion of a name on this list is not to be construed as taxonomic acceptance of the taxon to which the name is applied. Indeed, some of these names may, in time, be shown to be synonyms, or the organisms may be transferred to another genus, thus necessitating the creation of a new combination. PMID- 15545407 TI - Notification that new names and new combinations have appeared in volume 54, part 4, of the IJSEM. AB - This listing of names published in a previous issue of the IJSEM is provided as a service to bacteriology to assist in the recognition of new names and new combinations. This procedure was proposed by the Judicial Commission [Minute 11(ii), Int J Syst Bacteriol 41 (1991), p. 185]. The names given herein are listed according to the Rules of priority (i.e. page number and order of valid publication of names in the original articles). Taxonomic opinions included in this List (i.e. the creation of synonyms or the emendation of circumscriptions) cannot be considered as validly published nor, in any other way, approved by the International Committee on Systematics of Prokaryotes and its Judicial Commission. PMID- 15545408 TI - Policy proposal for publication of papers with data sets from genome-wide studies. PMID- 15545409 TI - An open letter on microarray data from the MGED Society. PMID- 15545410 TI - Methane utilization by Methylobacterium species: new evidence but still no proof for an old controversy. PMID- 15545411 TI - Chromohalobacter sarecensis sp. nov., a psychrotolerant moderate halophile isolated from the saline Andean region of Bolivia. AB - A moderately halophilic, aerobic, motile, Gram-negative, rod-shaped bacterium (strain LV4(T)) was isolated from saline soil around the lake Laguna Verde in the Bolivian Andes. The organism is a heterotroph, able to utilize various carbohydrates as a carbon source. It showed tryptophan deaminase, oxidase and catalase activity, but was unable to produce indole or H(2)S; nitrate was not reduced. The G+C content of the genomic DNA was 56.1 mol%. The pH range for growth was 5-10, temperature range was 0-45 degrees C and the range of NaCl concentrations was 0-25 % (w/v). On the basis of 16S rRNA gene sequence analysis, strain LV4(T) was found to be closely related to Chromohalobacter canadensis DSM 6769(T) and Pseudomonas beijerinckii DSM 7218(T); however, its DNA-DNA relatedness with these type strains was low. Strain LV4(T) resembled other Chromohalobacter species with respect to various physiological, biochemical and nutritional characteristics but also exhibited differences. Thus, a novel species, Chromohalobacter sarecensis sp. nov., is proposed, with LV4(T) (=CCUG 47987(T)=ATCC BAA-761(T)) as the type strain. PMID- 15545412 TI - Nocardia shimofusensis sp. nov., isolated from soil, and Nocardia higoensis sp. nov., isolated from a patient with lung nocardiosis in Japan. AB - Three actinomycete strains isolated from soils and one strain isolated from a patient with lung nocardiosis in 1999 and 2001 in Japan have been provisionally assigned to the genus Nocardia on the basis of morphological criteria. These isolates were further investigated to determine their specific taxonomic status. Detailed chemotaxonomic characterization and 16S rRNA gene sequence analysis of these isolates confirmed that they belong to the genus Nocardia. The 16S rRNA gene sequences of the four strains were most similar to that of Nocardia farcinica. However, the sequence similarity values between these four strains and N. farcinica were <98.9 %. These four strains were susceptible to 5-fluorouracil, and they have the ability to decompose urea, which is a very characteristic trait. Furthermore, DNA-DNA relatedness data revealed that IFM 10311(T), IFM 10312 and IFM 10313 comprise a single novel species of Nocardia, that IFM 10084(T) represents another novel species of Nocardia and that these two novel species could be distinguished from N. farcinica. The names Nocardia shimofusensis sp. nov. and Nocardia higoensis sp. nov. are proposed, with IFM 10311(T) (=NBRC 100134(T)=JCM 12122(T)=DSM 44733(T)) and IFM 10084(T) (=NBRC 100133(T)=JCM 12121(T)=DSM 44732(T)) as the respective type strains. PMID- 15545413 TI - Caenibacterium thermophilum is a later synonym of Schlegelella thermodepolymerans. AB - Recently, two strains of Schlegelella thermodepolymerans Elbanna et al. 2003 and an independently isolated bacterium, Caenibacterium thermophilum Manaia et al. 2003, were described in parallel as gen. nov., sp. nov. Analysis of the 16S rRNA genes revealed similarity between C. thermophilum and the two strains of S. thermodepolymerans of 99.8 and 99.6 %, respectively. DNA-DNA hybridization experiments revealed mean DNA reassociation levels of 97-98 % among C. thermophilum and the two strains of S. thermodepolymerans, thereby confirming the close relationship and indicating that C. thermophilum is a later synonym of S. thermodepolymerans. PMID- 15545414 TI - Genome reduction in prokaryotic obligatory intracellular parasites of humans: a comparative analysis. AB - Obligatory intracellular parasites have undergone significant genome reduction by gene loss over time in the context of their obligate associations with the host. The flux, streamlining and elimination of genes in these genomes constitute a selective and ongoing process. Comparative analyses of five completely sequenced obligatory intracellular parasite genomes reveal that these genomes display marked similarities in patterns of protein length and frequency distribution, with substantial sharing of a 'backbone genome'. From category distribution based on the database of cluster of orthologous groups of proteins (COG), it is clear that habitat is a major factor contributing to genome reduction. It is also observed that, in all five obligatory intracellular parasites, the reduction in number of genes/proteins is greater for proteins with lengths of 200-600 amino acids. These comparative analyses highlight that gene loss is function-dependent, but is independent of protein length. These comparisons enhance our knowledge of the forces that drive the extreme specialization of the bacteria and their association with the host. PMID- 15545415 TI - Shewanella profunda sp. nov., isolated from deep marine sediment of the Nankai Trough. AB - A novel piezotolerant, mesophilic, facultatively anaerobic, organotrophic, polarly flagellated bacterium (strain LT13a(T)) was isolated from a deep sediment layer in the Nankai Trough (Leg 190, Ocean Drilling Program) off the coast of Japan. This organism used a wide range of organic substrates as sole carbon and energy sources: pyruvate, glutamate, succinate, fumarate, lactate, citrate, peptone and tryptone. Oxygen, nitrate, fumarate, ferric iron and cystine were used as electron acceptors. Maximal growth rates were observed at a hydrostatic pressure of 10 MPa. Hydrostatic pressure for growth was in the range 0.1-50 MPa. Predominant cellular fatty acids were 16 : 1omega7c, 15 : 0 iso, 16 : 0 and 13 : 0 iso. The G+C content of the DNA was 44.9 mol%. On the basis of 16S rRNA gene sequences, strain LT13a(T) was shown to belong to the gamma-Proteobacteria, being closely related to Shewanella putrefaciens (98 %), Shewanella oneidensis (97 %) and Shewanella baltica (96 %). Levels of DNA homology between strain LT13a(T) and S. putrefaciens, S. oneidensis and S. baltica were <20 %, indicating that strain LT13a(T) represents a novel species. Genetic evidence and phenotypic characteristics showed that isolate LT13a(T) constitutes a novel species of the genus Shewanella. Because of the deep origin of the strain, the name Shewanella profunda sp. nov. is proposed, with LT13a(T) (=DSM 15900(T)=JCM 12080(T)) as the type strain. PMID- 15545416 TI - Reclassification of Mesoplasma pleciae as Acholeplasma pleciae comb. nov. on the basis of 16S rRNA and gyrB gene sequence data. AB - Genomic DNA sequence data for the 16S rRNA gene and the gyrB gene of Mesoplasma pleciae PS-1(T) (=ATCC 49582(T)=NBRC 100476(T)) demonstrate a much closer relationship to Acholeplasma laidlawii and Acholeplasma oculi than to other species in the order Entomoplasmatales. In addition, the preferred use of UGG rather than UGA as the codon for tryptophan in the gyrB sequence probably places the organism outside the order Entomoplasmatales. It is proposed that M. pleciae be reclassified in the genus Acholeplasma, as Acholeplasma pleciae comb. nov. PMID- 15545417 TI - Thermosipho atlanticus sp. nov., a novel member of the Thermotogales isolated from a Mid-Atlantic Ridge hydrothermal vent. AB - A novel anaerobic, thermophilic and heterotrophic bacterium, designated strain DV1140(T), was isolated from a deep-sea hydrothermal vent sample from the Mid Atlantic Ridge. The cells were non-motile straight rods, 1.8 microm long and 0.4 microm wide, surrounded by an outer sheath-like structure (toga). They grew at 45 80 degrees C (optimum 65 degrees C), pH 5.0-9.0 (optimum pH 6.0) and at sea salt concentrations of 20-60 g l(-1) (optimum 30 g l(-1)). Strain DV1140(T) was able to ferment yeast extract, peptone, brain heart infusion, gelatin, starch, galactose, arabinose, glucose, trehalose and cellobiose. The fermentation products identified on glucose in the presence of yeast extract and peptone were acetate, isovalerate and hydrogen. The G+C content of the genomic DNA was 33 mol%. Phylogenetic analysis of the 16S rRNA gene sequence (GenBank accession number AJ577471) located the strain within the genus Thermosipho in the bacterial domain. On the basis of 16S rRNA gene sequence comparisons, and physiological and biochemical characteristics, the isolate represents a novel species, for which the name Thermosipho atlanticus sp. nov. is proposed. The type strain is DV1140(T) (=CIP 108053(T)=DSM 15807(T)). PMID- 15545418 TI - Diversity of bartonellae associated with small mammals inhabiting Free State province, South Africa. AB - The prevalence and diversity of bartonellae infecting the blood of 10 small mammal species inhabiting nine Nature Reserves of the Free State province, South Africa, was assessed using phenotypic, genotypic and phylogenetic methods. Of 86 small mammals sampled, 38 animals belonging to five different species yielded putative bartonellae. Thirty-two isolates were confirmed as bartonellae and were characterized by comparison of partial citrate synthase gene (gltA) sequences. Phylogenetic reconstructions derived from alignment of these sequences with those available for other bartonellae indicated that the South African rodent associated isolates formed two distinct clades within the radius of the genus Bartonella. One of these clades also included recognized Bartonella species associated with rodents native to Eurasia but not to the New World, whereas the second clade contained exclusively isolates associated with South African rodents. Comparison of gltA sequences delineated the isolates into a number of ecologically distinct populations and provided an indication that a combination of phylogenetics and the identification of sequence clusters in housekeeping protein-encoding genes could be developed as a key criterion in the classification of bartonellae. This study is the first to investigate wildlife associated bartonellae in Africa, adding support to their ubiquity and broad diversity and to the paradigm that the phylogenetic positions of the Bartonella species encountered today have been influenced by the geographical distribution of their hosts. PMID- 15545419 TI - Algoriphagus antarcticus sp. nov., a novel psychrophile from microbial mats in Antarctic lakes. AB - A taxonomic study was performed on six strains isolated from microbial mats of lakes Reid, Fryxell and Ace in Antarctica. Phylogenetic analysis based on 16S rRNA gene sequences indicated that these strains belonged to the family 'Flexibacteraceae' and were closely related to the recently described genera Algoriphagus and Hongiella. The isolates were Gram-negative, chemoheterotrophic, aerobic, psychrophilic, orange-red-pigmented bacteria and their DNA G+C content ranged from 39.9 to 41.0 mol%. Whole-cell fatty acid profiles included mainly branched fatty acids and summed feature 3, comprising 15 : 0 iso 2OH, 16 : 1omega7c or both. On the basis of genotypic, phenotypic, chemotaxonomic and phylogenetic results, the novel strains were classified as Algoriphagus antarcticus sp. nov. The type strain is LMG 21980(T) (=DSM 15986(T)=R-10710(T)). PMID- 15545420 TI - Janibacter melonis sp. nov., isolated from abnormally spoiled oriental melon in Korea. AB - Two Gram-positive bacterial strains, CM2104(T) and CM2110, isolated from the inner part of abnormally spoiled oriental melon (Cucumis melo) in Korea, were subjected to a polyphasic taxonomic study. The cell-wall peptidoglycan of strains CM2104(T) and CM2110 contained meso-diaminopimelic acid as the diagnostic diamino acid. The predominant menaquinone was MK-8(H(4)). The major fatty acids detected in the two strains were iso-C(16 : 0), C(17 : 1)omega8c and C(18 : 1)omega9c or C(17 : 0). The DNA G+C content of the two strains was 73 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strains formed a coherent cluster with a clade comprising two Janibacter species, Janibacter limosus and Janibacter terrae. Strains CM2104(T) and CM2110 exhibited a 16S rRNA gene sequence similarity value of 99.7 % and a mean DNA-DNA relatedness level of 89 %. Strains CM2104(T) and CM2110 showed 16S rRNA gene sequence similarity levels of 97.8-98.4 % to the type strains of J. limosus and J. terrae. DNA-DNA relatedness between strains CM2104(T) and CM2110 and the type strains of these two Janibacter species was 7-11 %. On the basis of the phenotypic and phylogenetic data and genomic distinctiveness, strains CM2104(T) and CM2110 should be placed within the genus Janibacter as members of a novel species, for which the name Janibacter melonis sp. nov. is proposed. The type strain is CM2104(T) (=KCTC 9987(T)=DSM 16063(T)=JCM 12321(T)). PMID- 15545421 TI - Erythrobacter aquimaris sp. nov., isolated from sea water of a tidal flat of the Yellow Sea in Korea. AB - Three Gram-negative, non-motile, non-spore-forming, slightly halophilic rods (strains SW-110(T), SW-116 and SW-140) were isolated from sea water of a tidal flat of the Yellow Sea in Korea and subjected to a polyphasic taxonomic study. The three isolates did not produce bacteriochlorophyll a and were characterized chemotaxonomically by having ubiquinone-10 as the predominant respiratory lipoquinone and C(18 : 1)omega7c and C(17 : 1)omega6c as the major fatty acids. The DNA G+C content of the three isolates was between 62.2 and 62.9 mol%. Strains SW-110(T), SW-116 and SW-140 showed no difference in their 16S rRNA gene sequences, and their mean level of DNA-DNA relatedness was 94.8 %. Phylogenetic analyses based on 16S rRNA gene sequences showed that the three strains form a distinct phylogenetic lineage within the cluster comprising Erythrobacter species. Similarities between the 16S rRNA gene sequences of strains SW-110(T), SW-116 and SW-140 and the type strains of Erythrobacter species ranged from 98.4 % (with Erythrobacter longus) to 97.7 % (with Erythrobacter flavus). Levels of DNA-DNA relatedness between strains SW-110(T), SW-116 and SW-140 and the type strains of all recognized Erythrobacter species were in the range 5.3-12.7 %. On the basis of polyphasic taxonomic data, strains SW-110(T), SW-116 and SW-140 were classified as a novel Erythrobacter species, for which the name Erythrobacter aquimaris sp. nov. is proposed. The type strain is SW-110(T) (=KCCM 41818(T)=JCM 12189(T)). PMID- 15545422 TI - Agromyces ulmi sp. nov., a xylanolytic bacterium isolated from Ulmus nigra in Spain. AB - Two xylan-degrading bacterial strains were isolated from a decayed Ulmus nigra tree in Spain. The isolates were Gram-positive, non-motile, aerobic and formed substrate mycelium which fragmented into irregular rods. 16S rRNA gene sequence analysis indicated that the isolates form a separate branch within the genus Agromyces phylogenetic cluster, with Agromyces mediolanus DSM 20152(T) being their closest relative (97.7 and 97.6 % sequence similarity). Catalase, nitrate reduction and urease tests differentiated these strains from A. mediolanus. Cell wall peptidoglycan composition, major menaquinone, predominant fatty acids and phospholipid pattern were typical of the genus Agromyces. The DNA G+C content determined for the type strain XIL01(T) was 72 mol%. Based on the data presented, a novel species Agromyces ulmi sp. nov. is proposed. The type strain is XIL01(T) (=LMG 21954(T)=DSM 15747(T)). PMID- 15545423 TI - Geobacillus lituanicus sp. nov. AB - Obligately thermophilic, aerobic, proteolytic, endospore-forming strain N-3(T) was isolated from a high-temperature oilfield in Lithuania. 16S rRNA gene sequence analysis placed this strain in genetic group 5 of the endospore formers. Geobacillus thermoleovorans appeared to be the closest phylogenetic neighbour (99.4 % sequence similarity). The G+C content of strain N-3(T) was 52.5 mol% and matched the range established for the genus Geobacillus. Studies of DNA-DNA relatedness and morphological and physiological analyses enabled strain N-3(T) to be described as a member of the genus Geobacillus, but could not assign this strain to any other known species of this genus. Results of this polyphasic study allowed characterization of strain N-3(T) as a novel species in the genus Geobacillus - Geobacillus lituanicus sp. nov. This species can be distinguished from G. thermoleovorans and Geobacillus stearothermophilus on the basis of 16S rRNA gene PCR-RFLP assays with the restriction endonucleases AluI, HaeIII and TaqI. The type strain of the novel species is N-3(T) (=DSM 15325(T)=VKM B 2294(T)). PMID- 15545424 TI - Bacillus asahii sp. nov., a novel bacterium isolated from soil with the ability to deodorize the bad smell generated from short-chain fatty acids. AB - In a screening campaign to isolate strains with the ability to remove the bad smell associated with animal faeces, strain MA001(T) was isolated from a soil sample obtained from Shizuoka prefecture, Japan. The isolate grew at pH 6-9 but not at pH 10. Cells were Gram-positive, straight rods with peritrichous flagella and produced ellipsoidal spores. The isolate was positive for catalase and oxidase tests but negative for indole production, deamination of phenylalanine and H(2)S production. The isolate did not produce acid from any carbohydrates tested and could not grow in more than 2 % NaCl. The DNA G+C content was 39.4 mol%. The cellular fatty acids profile consisted of significant amount of C(15) branched-chain fatty acids, iso-C(15 : 0) and anteiso-C(15 : 0). Phylogenetic analysis based on 16S rRNA gene sequencing indicated that strain MA001(T) was closely related to Bacillus simplex and Bacillus psychrosaccharolyticus. DNA-DNA hybridization revealed a low relatedness of the isolate to several phylogenetically close neighbours (less than 9 %). On the basis of the phenotypic characteristics observed, phylogenetic data based on 16S rRNA gene sequencing and DNA-DNA relatedness data, it is concluded that the isolate should be classified as representing a novel species, for which the name Bacillus asahii is proposed. The type strain is MA001(T) (=JCM 12112(T)=NCIMB 13969(T)). PMID- 15545425 TI - Mesorhizobium septentrionale sp. nov. and Mesorhizobium temperatum sp. nov., isolated from Astragalus adsurgens growing in the northern regions of China. AB - Ninety-five rhizobial strains isolated from Astragalus adsurgens growing in the northern regions of China were classified into three main groups, candidate species I, II and III, based on a polyphasic approach. Comparative analysis of full-length 16S rRNA gene sequences of representative strains showed that candidate species I and II were Mesorhizobium, while candidate species III, which consisted of non-nodulating strains, was closely related to Agrobacterium tumefaciens. The phylogenetic relationships of the three candidate species and some related strains were also confirmed by the sequencing of glnA genes, which were used as an alternative chromosomal marker. The DNA-DNA relatedness was between 11.3 and 47.1 % among representative strains of candidate species I and II and the type strains of defined Mesorhizobium species. Candidate III had DNA relatedness of between 4.3 and 25.2 % with type strains of Agrobacterium tumefaciens and Agrobacterium rubi. Two novel species are proposed to accommodate candidate species I and II, Mesorhizobium septentrionale sp. nov. (type strain, SDW014(T)=CCBAU 11014(T)=HAMBI 2582(T)) and Mesorhizobium temperatum sp. nov. (type strain, SDW018(T)=CCBAU 11018(T)=HAMBI 2583(T)), respectively. At least two distinct nodA sequences were identified among the strains. The numerically dominant nodA sequence type was most similar to that from the Mesorhizobium tianshanense type strain and was identified in strains belonging to the two novel species as well as other, as yet, undefined genome types. Host range studies indicate that the different nodA sequences correlate with different host ranges. Further comparative studies with the defined Agrobacterium species are needed to clarify the taxonomic identity of candidate species III. PMID- 15545426 TI - Exiguobacterium oxidotolerans sp. nov., a novel alkaliphile exhibiting high catalase activity. AB - A novel alkaliphile was isolated from a drain of a fish processing plant. The isolate grew at a pH range of 7-10. Cells were Gram-positive, facultatively aerobic, motile rods with peritrichous flagella. Colonies were orange or yellow in colour. Catalase and oxidase reactions were positive. The isolate grew in 0-12 % NaCl but not above 15 % NaCl. Its cell extract exhibited 567 times higher catalase activity than an Escherichia coli cell extract. The major cellular fatty acids were iso-C(13 : 0), anteiso-C(13 : 0), iso-C(15 : 0), iso-C(16 : 0), iso C(17 : 0), anteiso-C(17 : 0) and iso-C(17 : 1). Its DNA G+C content was 46.7 mol%. Phylogenetic analysis based on 16S rRNA gene sequencing and chemotaxonomic data indicated that strain T-2-2(T) is a member of the genus Exiguobacterium. DNA DNA hybridization revealed a low relatedness of the isolate to several phylogenetic neighbours (less than 25 %). On the basis of phenotypic characteristics, phylogenetic data and DNA-DNA relatedness data, the isolate merits classification as a novel species, for which the name Exiguobacterium oxidotolerans sp. nov. is proposed. The type strain is T-2-2(T) (=JCM 12280(T)=NCIMB 13980(T)). PMID- 15545427 TI - Geobacillus gargensis sp. nov., a novel thermophile from a hot spring, and the reclassification of Bacillus vulcani as Geobacillus vulcani comb. nov. AB - A novel thermophilic spore-forming strain, Ga(T), was isolated from the Garga hot spring located in the northern part of the Transbaikal region (Russia). Strain Ga(T) was found to be an aerobic, Gram-positive, rod-shaped, thermophilic (optimum growth temperature is 60-65 degrees C), chemo-organotrophic bacterium that grows on various sugars, carboxylic acids and hydrocarbons. The G+C content of its DNA is 52.9 mol%. The 16S rRNA gene sequence similarity data show that strain Ga(T) is closely related to members of the genus Geobacillus. Relevant chemotaxonomic data (in particular, the major fatty acid profile of strain Ga(T), which includes iso-C15 : 0, iso-C16 : 0 and iso-C17 : 0 acids) support the assignment of this strain to the genus Geobacillus. The physiological, biochemical and DNA-DNA hybridization studies of strain Ga(T) showed that it differs both genotypically and phenotypically from the recognized Geobacillus species. Based on these data, strain Ga(T) belongs to a novel species, Geobacillus gargensis sp. nov. (type strain, Ga(T)=VKM B-2300(T)=DSM 15378(T)). The analysis of the phenotypic characteristics (additional to those given in the original description) of the type strain of Bacillus vulcani (DSM 13174(T)) showed that they are very similar to the major phenotypic characteristics of the genus Geobacillus. The low DNA-DNA reassociation values of strain DSM 13174(T) with various species of this genus (from 38 to 54 %) clearly demonstrate a sufficient genomic distinction of this strain and its taxonomic status as a species. The physiological characteristics, phylogenetic position and DNA-DNA reassociation values of B. vulcani allow this species to be reclassified as Geobacillus vulcani comb. nov. The main properties that differentiate G. vulcani from the other species of the genus are its ability to produce acids from glycerol, lactose and ribose. PMID- 15545428 TI - Nocardia araoensis sp. nov. and Nocardia pneumoniae sp. nov., isolated from patients in Japan. AB - Two actinomycete strains isolated from two patients with lung nocardiosis between 1995 and 1997 in Japan were assigned to novel species of the genus Nocardia based on morphological and chemical criteria. Comparative 16S rRNA gene sequence analysis of the two strains revealed that they belong to the genus Nocardia and are most closely related to the species Nocardia beijingensis. Determination of DNA-DNA relatedness indicated that these strains could be assigned to two novel species. Based on their phenotypic and phylogenetic characters, two novel species of the genus Nocardia are proposed: Nocardia araoensis sp. nov. for IFM 0575(T) (=NBRC 100135(T)=JCM 12118(T)=DSM 44729(T)) and Nocardia pneumoniae sp. nov. for IFM 0784(T) (=NBRC 100136(T)=JCM 12119(T)=DSM 44730(T)). PMID- 15545429 TI - Paenibacillus elgii sp. nov., with broad antimicrobial activity. AB - Two novel spore-forming bacteria with broad antimicrobial activity were isolated from roots of Perilla frutescens. The isolates, SD17(T) and SD18, were facultatively anaerobic and showed variable Gram reaction. Growth was observed between 20 and 45 degrees C. DNA G+C content of SD17(T) was 51.7 mol%, and the major fatty acid was anteiso-C(15 : 0) (54.1 %). 16S rRNA gene sequence similarity of SD17(T) ranged from 98.6 to 91.3 % with other Paenibacillus species. The phylogenetic tree showed that isolate SD17(T) formed a significant monophyletic clade with Paenibacillus koreensis KCTC 2393(T) and Paenibacillus ehimensis IFO 15659(T). DNA-DNA relatedness values for strain SD17(T) with Paenibacillus koreensis KCTC 2393(T) and Paenibacillus ehimensis IFO 15659(T) were 17.4 and 19.8 %, respectively. These isolates thus merit species status within Paenibacillus, for which the name Paenibacillus elgii sp. nov. is proposed. The type strain is SD17(T) (=KCTC 10016BP(T)=NBRC 100335(T)). PMID- 15545430 TI - Halomonas koreensis sp. nov., a novel moderately halophilic bacterium isolated from a solar saltern in Korea. AB - A moderately halophilic bacterium, strain SS20(T), capable of growing at salinities of 1-20 % (w/v) NaCl was isolated from a solar saltern of the Dangjin area in Korea and was characterized taxonomically. Strain SS20(T) was a Gram negative bacterium comprising motile, short rods. Its major cellular fatty acids were C(18 : 1)omega7c, C(19 : 0)omega8c cyclo and C(16 : 0). The DNA G+C content was 70 mol% and the predominant ubiquinone was Q-9. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SS20(T) belonged to the genus Halomonas. The levels of 16S rRNA gene sequence similarity to the type strains of Halomonas species were in the range 93.0-97.5 %. The levels of DNA-DNA relatedness between strain SS20(T) and the type strains of phylogenetically closely related Halomonas species were in the range 5.3-12.3 %. On the basis of physiological and molecular properties, strain SS20(T) represents a novel species of the genus Halomonas, for which the name Halomonas koreensis sp. nov. is proposed. The type strain is SS20(T) (=KCTC 12127(T)=JCM 12237(T)). PMID- 15545431 TI - Clostridium straminisolvens sp. nov., a moderately thermophilic, aerotolerant and cellulolytic bacterium isolated from a cellulose-degrading bacterial community. AB - A novel anaerobic, thermophilic and cellulolytic bacterium (strain CSK1(T)) was isolated from a cellulose-degrading bacterial community. On the basis of 16S rRNA gene sequence similarity, strain CSK1(T) was mapped to cluster III of the genus Clostridium. Strain CSK1(T) is closely related to Clostridium thermocellum (96.2 %) and Clostridium aldrichii (95.1 %). Strain CSK1(T) is a non-motile, spore forming, straight or slightly curved rod. The optimum temperature and initial pH for its growth and cellulose degradation are 50-55 degrees C and pH 7.5. Strain CSK1(T) grew under a gas phase containing up to 4 % O(2). Phylogenetic and phenotypic analyses support the differentiation of strain CSK1(T) from its closest relatives. Strain CSK1(T) therefore represents a novel species, for which the name Clostridium straminisolvens sp. nov. is proposed, with CSK1(T) (=DSM 16021(T)=IAM 15070(T)) as the type strain. PMID- 15545432 TI - Reclassification of Sphaerobacter thermophilus from the subclass Sphaerobacteridae in the phylum Actinobacteria to the class Thermomicrobia (emended description) in the phylum Chloroflexi (emended description). AB - Sphaerobacter thermophilus was originally classified as the deepest branching member of the phylum Actinobacteria (high-G+C, Gram-positive bacteria) based on 16S rRNA gene comparative analysis. However, the analysis lacked suitable outgroups, and additional 16S rRNA gene sequences indicate that it is most closely related to Thermomicrobium roseum, which it also resembles phenotypically. Furthermore, both species are reproducibly affiliated with the phylum Chloroflexi (green non-sulfur bacteria), despite T. roseum currently being classified in its own phylum, the Thermomicrobia. Transfer of Sphaerobacter to the class Thermomicrobia, and transfer of the class Thermomicrobia to the phylum Chloroflexi, are proposed. Descriptions for the phylum Chloroflexi and the class Thermomicrobia are emended to reflect the proposed changes in classification. PMID- 15545433 TI - Planotetraspora silvatica sp. nov. and emended description of the genus Planotetraspora. AB - An actinomycete that developed sporangia containing four spores in a single row at the ends of short sporangiophores on branched aerial hyphae was isolated from subtropical forest soil. The isolate contained menaquinone MK-9(H(4)), glutamic acid, alanine and meso-diaminopimelic acid as cell-wall amino acids and madurose in the whole-cell hydrolysate. The 16S rRNA gene sequence indicated that the isolate formed a monophyletic cluster with Planotetraspora mira. On the basis of morphological and chemotaxonomic characteristics, phylogenetic analysis and DNA DNA relatedness data, a novel species of the genus Planotetraspora is proposed, Planotetraspora silvatica sp. nov. (type strain, TT 00-51(T)=NBRC 100141(T)=DSM 44746(T)). PMID- 15545434 TI - Mycobacterium nebraskense sp. nov., a novel slowly growing scotochromogenic species. AB - The characterization of a novel slowly growing, scotochromogenic Mycobacterium species is reported. This previously undescribed mycobacterial species was isolated from five different patients with symptomatic pulmonary infections. All isolates were acid-fast-positive and the mycolic acid profiles were unique and supported placement into the genus Mycobacterium. Phenotypic characteristics of each strain included optimal growth after 3 weeks at a temperature range of 30-35 degrees C, yellow pigmentation after incubation in the dark and production of a heat-stable catalase. The 16S rRNA gene and internal transcribed spacer 1 sequences were identical for all five strains, but distinct from all known mycobacterial species. Phylogenetic analysis based on the 16S rRNA gene sequence placed the novel species within the slowly growing mycobacteria group in close proximity to Mycobacterium malmoense, Mycobacterium avium, Mycobacterium kansasii and Mycobacterium scrofulaceum. These data support the conclusion that the related five described organisms represent a novel Mycobacterium species, for which the name Mycobacterium nebraskense sp. nov. is proposed, with the type strain UNMC-MY1349(T) (=ATCC BAA-837(T)=DSM 44803(T)). PMID- 15545435 TI - Streptomyces ferralitis sp. nov., a novel streptomycete isolated from a New Caledonian ultramafic soil. AB - The taxonomic position of an actinomycete isolated from an ultramafic soil in New Caledonia was determined using a polyphasic approach. The isolate, which was designated SFOp68(T), was shown to have chemical and morphological properties typical of streptomycetes. An almost complete 16S rRNA gene sequence of the isolate was generated and compared with sequences of representative streptomycetes. The 16S rRNA data not only supported the classification of the strain in the genus Streptomyces, but also showed that it formed a distinct phyletic line that was most closely related to one composed of the type strain of Streptomyces rimosus. The two organisms can be readily separated using a diverse range of phenotypic properties. It is proposed that strain SFOp68(T) (=DSM 41836(T)=NCIMB 13954(T)) be classified in the genus Streptomyces as Streptomyces ferralitis sp. nov. PMID- 15545436 TI - Arthrobacter psychrophenolicus sp. nov., isolated from an alpine ice cave. AB - On the basis of phenotypic, genotypic and chemotaxonomic characteristics, a novel species belonging to the genus Arthrobacter is described. A facultatively psychrophilic bacterium, strain AG31(T), was isolated from an alpine ice cave. The aerobic, Gram-positive, non-spore-forming, non-motile strain exhibited a rod coccus growth cycle and produced a yellow pigment. Good growth and phenol biodegradation occurred at a temperature range of 1-25 degrees C. Up to 10 mM phenol was utilized as a sole carbon source. Glucose was not assimilated. Analysis of the 16S rRNA gene revealed that strain AG31(T) represents a distinct lineage within the genus Arthrobacter, being most closely related to Arthrobacter sulfureus. The level of DNA-DNA relatedness to the type strain of A. sulfureus was 29.9 %. Anteiso-C(15 : 0) was the predominant fatty acid (72 %). Strain AG31(T) exhibited A4alpha l-lys-l-Glu-type peptidoglycan and contained glucose as the only cell-wall sugar. MK-10 was the predominant menaquinone, and the polar lipid pattern consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol and an unidentified glycolipid. Strain AG31(T) (=DSM 15454(T)=LMG 21914(T)) is assigned as the type strain of a novel Arthrobacter species, Arthrobacter psychrophenolicus sp. nov. PMID- 15545437 TI - Aeromonas molluscorum sp. nov., isolated from bivalve molluscs. AB - Five Aeromonas strains (848T(T), 93M, 431E, 849T and 869N), which were isolated from bivalve molluscs and were recognized previously by numerical taxonomy as members of an unknown Aeromonas taxon, were subjected to a polyphasic taxonomic study. DNA-DNA hybridization experiments showed that DNA of strain 848T(T) was <70 % similar (27-45 %) to that of the type/reference strains of the current Aeromonas hybridization groups (HGs), but 93 % similar to that of strain 93M. The DNA G+C content of the five strains ranged from 59.0 to 59.4 mol%. 16S rRNA gene sequence analysis confirmed that the strains belonged to the genus Aeromonas and showed high similarity to Aeromonas encheleia. Amplified fragment length polymorphism fingerprinting clustered the novel strains in a homogeneous group with low genotypic relatedness to other Aeromonas species. Useful phenotypic features for differentiating the five isolates from other Aeromonas species include their negative reactions in tests for indole production, lysine decarboxylase, gas from glucose and starch hydrolysis. From the results of this study, the name Aeromonas molluscorum sp. nov. is proposed for these strains, with the type strain 848T(T) (=CECT 5864(T)=LMG 22214(T)). PMID- 15545438 TI - Hydrogenivirga caldilitoris gen. nov., sp. nov., a novel extremely thermophilic, hydrogen- and sulfur-oxidizing bacterium from a coastal hydrothermal field. AB - A novel extremely thermophilic, hydrogen- and sulfur-oxidizing bacterium, designated strain IBSK3(T), was isolated from a coastal hot spring in Ibusuki, Kagoshima Prefecture, Japan. The cells were motile, straight to slightly curved rods (1.2-3.0 microm long and 0.3-0.4 microm wide). Strain IBSK3(T) was an obligate chemolithoautotroph growing by respiratory nitrate reduction with H(2), forming N(2)O as an end product. Low concentrations of O(2) (0.4-7.7 %, v/v; optimum 2.0 %, v/v) could serve as an alternative electron acceptor to growth. In addition, strain IBSK3(T) was able to utilize elemental sulfur as a sole electron donor with either nitrate or low concentrations of O(2) as an electron acceptor. Growth was observed between 55 and 77.5 degrees C (optimum 75 degrees C; 2 h doubling time), pH 5.5 and 8.3 (optimum pH 6.5-7.0), and in the presence of 0.5 and 4.0 % NaCl (optimum 2.0 %). The G+C content of the genomic DNA was 49.2 mol%. On the basis of 16S rRNA gene sequence analysis, strain IBSK3(T) belonged to the family Aquificaceae, but it only demonstrated a distant phylogenetic relationship with any recognized species within the family (sequence similarity was less than 92 %). On the basis of the physiological and molecular characteristics of the novel isolate, a new genus and novel species are proposed: the type strain of Hydrogenivirga caldilitoris gen. nov., sp. nov. is IBSK3(T) (=JCM 12173(T)=ATCC BAA-821(T)). PMID- 15545439 TI - Streptomyces glauciniger sp. nov., a novel mesophilic streptomycete isolated from soil in south China. AB - A polyphasic study was undertaken to establish the taxonomic status of a soil isolate. The organism, strain FXJ14(T), was found to have chemical and morphological properties characteristic of streptomycetes. Phylogenetic analyses based on an almost complete 16S rRNA gene sequence of the strain and on the 120 nt variable gamma-region of the 16S rRNA molecule showed that it formed a distinct phyletic line within the range of variation encompassed by the genus Streptomyces. The sharp separation of the organism from representatives of the genus Streptomyces was strengthened by the fact that its BOX-PCR and RFLP of 16S rDNA-ITS fingerprints differed from those of over 450 recognized Streptomyces species. The isolate also had a profile of phenotypic properties that readily distinguished it from the genotypically close type strains. It is evident from the combination of genotypic and phenotypic data that strain FXJ14(T) (=AS 4.1858(T)=JCM 12278(T)=LMG 22082(T)) should be classified as the type strain of a novel species of the genus Streptomyces, for which the name Streptomyces glauciniger sp. nov. is proposed. PMID- 15545440 TI - Saccharothrix xinjiangensis sp. nov., a pyrene-degrading actinomycete isolated from Tianchi Lake, Xinjiang, China. AB - A pyrene-degrading isolate, strain PYX-6(T), was obtained from Tianchi Lake, Xinjiang Uygur Autonomous Region, China, and was identified by means of polyphasic taxonomy. On the basis of 16S rRNA gene sequence similarity, strain PYX-6(T) was closely related to members of the genus Saccharothrix. Its 16S rRNA gene sequence similarity to each of Saccharothrix syringae, Saccharothrix australiensis, Saccharothrix algeriensis and Saccharothrix coeruleofusca was 98 %. Chemotaxonomic and physiological properties of strain PYX-6(T) further supported its affiliation to the genus Saccharothrix. DNA-DNA hybridization results and physiological and biochemical tests allowed genotypic and phenotypic differentiation of strain PYX-6(T) from recognized Saccharothrix species. Therefore, strain PYX-6(T) represents a novel species, for which the name Saccharothrix xinjiangensis sp. nov. is proposed, with type strain PYX-6(T) (=AS 4.1731(T)=JCM 12329(T)). PMID- 15545441 TI - Dissection of phylogenetic relationships among 19 rapidly growing Mycobacterium species by 16S rRNA, hsp65, sodA, recA and rpoB gene sequencing. AB - The current classification of non-pigmented and late-pigmenting rapidly growing mycobacteria (RGM) capable of producing disease in humans and animals consists primarily of three groups, the Mycobacterium fortuitum group, the Mycobacterium chelonae-abscessus group and the Mycobacterium smegmatis group. Since 1995, eight emerging species have been tentatively assigned to these groups on the basis of their phenotypic characters and 16S rRNA gene sequence, resulting in confusing taxonomy. In order to assess further taxonomic relationships among RGM, complete sequences of the 16S rRNA gene (1483-1489 bp), rpoB (3486-3495 bp) and recA (1041 1056 bp) and partial sequences of hsp65 (420 bp) and sodA (441 bp) were determined in 19 species of RGM. Phylogenetic trees based upon each gene sequence, those based on the combined dataset of the five gene sequences and one based on the combined dataset of the rpoB and recA gene sequences were then compared using the neighbour-joining, maximum-parsimony and maximum-likelihood methods after using the incongruence length difference test. Combined datasets of the five gene sequences comprising nearly 7000 bp and of the rpoB+recA gene sequences comprising nearly 4600 bp distinguished six phylogenetic groups, the M. chelonae-abscessus group, the Mycobacterium mucogenicum group, the M. fortuitum group, the Mycobacterium mageritense group, the Mycobacterium wolinskyi group and the M. smegmatis group, respectively comprising four, three, eight, one, one and two species. The two protein-encoding genes rpoB and recA improved meaningfully the bootstrap values at the nodes of the different groups. The species M. mucogenicum, M. mageritense and M. wolinskyi formed new groups separated from the M. chelonae-abscessus, M. fortuitum and M. smegmatis groups, respectively. The M. mucogenicum group was well delineated, in contrast to the M. mageritense and M. wolinskyi groups. For phylogenetic organizations derived from the hsp65 and sodA gene sequences, the bootstrap values at the nodes of a few clusters were <70 %. In contrast, phylogenetic organizations obtained from the 16S rRNA, rpoB and recA genes were globally similar to that inferred from combined datasets, indicating that the rpoB and recA genes appeared to be useful tools in addition to the 16S rRNA gene for the investigation of evolutionary relationships among RGM species. Moreover, rpoB gene sequence analysis yielded bootstrap values higher than those observed with recA and 16S rRNA genes. Also, molecular signatures in the rpoB and 16S rRNA genes of the M. mucogenicum group showed that it was a sister group of the M. chelonae-abscessus group. In this group, M. mucogenicum ATCC 49650(T) was clearly distinguished from M. mucogenicum ATCC 49649 with regard to analysis of the five gene sequences. This was in agreement with phenotypic and biochemical characteristics and suggested that these strains are representatives of two closely related, albeit distinct species. PMID- 15545442 TI - Brevibacterium celere sp. nov., isolated from degraded thallus of a brown alga. AB - Two whitish yellow, Gram-positive, non-motile, aerobic bacteria were isolated from enrichment culture during degradation of the thallus of the brown alga Fucus evanescens. The bacteria studied were chemo-organotrophic, mesophilic and grew well on nutrient media containing up to 15 % (w/v) NaCl. The DNA G+C content was 61 mol%. The two isolates exhibited a conspecific DNA-DNA relatedness value of 98 %, indicating that they belong to the same species. A comparative analysis of 16S rRNA gene sequences revealed that strain KMM 3637(T) formed a distinct phyletic lineage in the genus Brevibacterium (family Brevibacteriaceae, class Actinobacteria) and showed the highest sequence similarity (about 97 %) to Brevibacterium casei. DNA-DNA hybridization experiments demonstrated 45 % binding with the DNA of B. casei DSM 20657(T). Physiological and chemotaxonomic characteristics (meso-diaminopimelic acid in the peptidoglycan, major cellular fatty acids 15 : 0ai and 17 : 0ai) of the bacteria studied were consistent with the genomic and phylogenetic data. On the basis of the results of this study, a novel species, Brevibacterium celere sp. nov., is proposed. The type strain is KMM 3637(T) (=DSM 15453(T)=ATCC BAA-809(T)). PMID- 15545443 TI - Transfer of Hyphomicrobium indicum to the genus Photobacterium as Photobacterium indicum comb. nov. AB - Hyphomicrobium indicum Johnson and Weisrock 1969 lacks true budding and hyphal branching, and some phenotypic characteristics are in contrast to other true hyphomicrobia. The major quinone system (ubiquinone Q-8), the G+C content of the DNA (40 mol%) and the cellular fatty acid composition (16 : 0, 16 : 1 and 18 : 1 as the major components, and 12 : 0 3-OH and 14 : 0 3-OH as the hydroxy fatty acids) of H. indicum are different from the genus Hyphomicrobium, but similar to the genus Photobacterium. Like the marine bacteria Photobacterium, H. indicum can be tolerant of sea water, while Hyphomicrobium cannot. Phylogenetic analyses of 16S rRNA and gyrB gene sequences revealed that H. indicum is most closely related to the genus Photobacterium of the gamma-Proteobacteria. Based on the phylogenetic, phenotypic and chemotaxonomic evidence, the results indicate that H. indicum should be transferred to the genus Photobacterium, and the name Photobacterium indicum comb. nov. (type strain, NBRC 14233(T)=ATCC 19614(T)) is proposed. PMID- 15545444 TI - Bacillus vietnamensis sp. nov., a moderately halotolerant, aerobic, endospore forming bacterium isolated from Vietnamese fish sauce. AB - Five strains of Gram-positive, endospore-forming, moderately halotolerant bacteria were studied taxonomically. Four were isolated from Vietnamese fish sauce and one from the Gulf of Mexico. Phylogenetic analysis based on 16S rRNA gene sequences showed that these strains clustered within the radiation of the genus Bacillus but separately from recognized Bacillus species. DNA G+C composition of the isolates ranged from 43 to 44 mol%. Strains 15-1(T) and NRRL B 14850 showed high levels of DNA-DNA relatedness (82-100 %) to each other and to the other strains isolated here; they displayed low levels of DNA-DNA relatedness (<29 %) to the type strains of selected recognized Bacillus species. They grew in 15 % NaCl and optimally in 1 % NaCl, which is characteristic of moderately halotolerant bacteria. The isolates grew at pH 6.5 to 10.0 but not at pH 6.0. Their cell walls contained meso-diaminopimelic acid. The major isoprenoid quinone was MK-7 and the principal cellular fatty acids were anteiso-C(15 : 0), iso-C(15 : 0) and anteiso-C(17 : 0). Based on these results, the strains tested were regarded as members of a novel Bacillus species for which the name Bacillus vietnamensis sp. nov. is proposed. The type strain is 15-1(T) (=JCM 11124(T)=NRIC 0531(T)=NRRL 23890(T)). PMID- 15545445 TI - Five novel Kitasatospora species from soil: Kitasatospora arboriphila sp. nov., K. gansuensis sp. nov., K. nipponensis sp. nov., K. paranensis sp. nov. and K. terrestris sp. nov. AB - A polyphasic study was carried out to establish the taxonomic positions of six strains isolated from diverse soil samples and provisionally assigned to the genus Kitasatospora. The isolates were found to have chemical and morphological properties consistent with their classification as Kitasatospora strains. Direct 16S rRNA gene sequence data confirmed the taxonomic position of the strains following the generation of phylogenetic trees by using three tree-making algorithms. Five of the isolates were considered to merit species status using complementary genotypic and phenotypic data. These organisms were designated Kitasatospora arboriphila sp. nov. (HKI 0189(T)=2291-120(T)=DSM 44785(T)=NCIMB 13973(T)), Kitasatospora gansuensis sp. nov. (HKI 0314(T)=2050-015(T)=DSM 44786(T)=NCIMB 13974(T)), Kitasatospora nipponensis sp. nov. (HKI 0315(T)=2148 013(T)=DSM 44787(T)=NCIMB 13975(T)), Kitasatospora paranensis sp. nov. (HKI 0190(T)=2292-041(T)=DSM 44788(T)=NCIMB 13976(T)) and Kitasatospora terrestris sp. nov. (HKI 0186(T)=2293-012(T)=DSM 44789(T)=NCIMB 13977(T)). The remaining organism, isolate HKI 0316 (=2122-022=DSM 44790=NCIMB 13978), was considered to be a strain of Kitasatospora kifunensis on the basis of 16S rRNA gene sequence, DNA-DNA relatedness and phenotypic data. PMID- 15545446 TI - Klebsiella singaporensis sp. nov., a novel isomaltulose-producing bacterium. AB - Cells of strain LX3(T), isolated from soil, were Gram-negative, facultatively anaerobic, non-motile, capsulated and non-endospore-forming straight rods, able to grow at 10 degrees C, unable to produce gas from lactose at 45 degrees C and unable to produce indole. The isolate converted sucrose to isomaltulose and did not produce detectable glucose by-products. The G+C content of the DNA was 56.4 mol%. Furthermore, comparison of 16S rRNA and rpoB gene sequences showed that the isolate clearly belongs to the genus Klebsiella. The closest phylogenetic relative was Klebsiella pneumoniae, there being 99.3 and 97.5 % similarity in 16S rRNA and rpoB gene sequences, respectively. DNA-DNA hybridization analysis demonstrated a very low level of relatedness to other members of the genus Klebsiella, indicating that the isolated strain and other species in the genus Klebsiella were not related at the species level. The isolate could be differentiated from other previously described members of the genus Klebsiella on the basis of phenotypic differences and 16S rRNA and rpoB gene sequence divergence, together with DNA-DNA reassociation data. Therefore, it is proposed that strain LX3(T) (=DSM 16265(T)=JCM 12419(T)) should be classified as the type strain of a novel species of genus Klebsiella, Klebsiella singaporensis sp. nov. PMID- 15545447 TI - Vibrio crassostreae sp. nov., isolated from the haemolymph of oysters (Crassostrea gigas). AB - Polyphasic analysis of five new Vibrio isolates originating from the haemolymph of diseased cultured oysters is described. The new isolates were closely related to Vibrio splendidus, having 98 % 16S rRNA gene sequence similarity. gyrB phylogenetic analysis, fluorescent amplified-fragment length polymorphism (FAFLP) fingerprinting and DNA-DNA hybridization experiments clearly showed that the new isolates form a tight genomic group that is different from the currently known Vibrio species. It is proposed to accommodate these isolates in a novel species, Vibrio crassostreae sp. nov. (type strain LGP 7(T)=LMG 22240(T)=CIP 108327(T)). Phenotypic and chemotaxonomic features that differentiate V. crassostreae from other known Vibrio species include arginine dihydrolase, utilization and fermentation of various carbon sources, beta-galactosidase activity, NO(2) production and the presence of the fatty acids 14 : 0 iso and 16 : 0 iso. PMID- 15545448 TI - Phenylobacterium lituiforme sp. nov., a moderately thermophilic bacterium from a subsurface aquifer, and emended description of the genus Phenylobacterium. AB - A facultative anaerobic bacterium, strain FaiI3(T), was isolated from samples collected from the free-flowing waters of a bore well (Fairlea Bore, registration number 3768) which taps into the Australian Great Artesian Basin subsurface thermal aquifer. Strain FaiI3(T) developed yellow to pale-yellow colonies (0.5 1.5 mm) after 48 h. The non-spore forming rods (0.5x1-3 microm) were slightly curved, occurred singly and as pairs and were motile with a single polar flagellum. Cells tended to form clumps in liquid medium and rosettes were commonly observed. The cells stained Gram-negative and electron micrographs of thin sections revealed a multi-layered complex Gram-negative cell wall structure. Strain FaiI3(T) grew optimally at 40-41 degrees C, with growth observed at 45 degrees C but not at 50 degrees C. The pH growth range was between pH 6 and 9 and optimal growth occurred between pH 6 and 6.5. Strain FaiI3(T) grew best with yeast extract as the sole carbon and energy source. Peptone, yeast extract, acetate, xylose, sucrose, glucose, glycerol, succinate, butyrate, lactate, fumarate, citrate, L-phenylalanine, cellobiose and gelatin supported growth but maltose, fructose, glycine, ethanol, benzoate and oxalate did not. Tyrosine was produced from L-phenylalanine. Strain FaiI3(T) was catalase-positive and oxidase negative and did not hydrolyse starch. Growth was inhibited by neomycin, tetracycline, streptomycin, chloramphenicol, ampicillin, vancomycin and spectinomycin. The G+C content was determined to be 66.5+/-0.5 mol%. On the basis of the 16S rRNA gene sequence analysis, strain FaiI3(T) was assigned as a novel species of the genus Phenylobacterium, Phenylobacterium lituiforme sp. nov. in the order Caulobacterales, subclass alpha-Proteobacteria, class Proteobacteria. The type strain is FaiI3(T) (=ATCC BAA-294(T)=DSM 14363(T)). PMID- 15545449 TI - Sphingomonas phyllosphaerae sp. nov., from the phyllosphere of Acacia caven in Argentina. AB - Two bacterial strains (FA1 and FA2(T)) were isolated from the phyllosphere of a leguminous tree, Acacia caven, in central Argentina. The strains were Gram negative, strictly aerobic, rod-shaped, motile and formed yellow-pigmented colonies on nutrient agar. The two-primer RAPD patterns of the two strains were identical, suggesting that they belong to the same species. The complete 16S rRNA gene sequences of the two strains were obtained and comparisons demonstrated that they cluster phylogenetically with the species of the genus Sphingomonas sensu stricto. Strain FA2(T) was most closely related (97.6 %) to Sphingomonas adhaesiva. 16S rRNA gene sequence similarities to all other established Sphingomonas species ranged from 94.4 % (to Sphingomonas echinoides) to 97.6 % (to S. adhaesiva). Strains FA1 and FA2(T) were catalase-positive and oxidase negative. Aesculin was hydrolysed, gelatin and urea were not. beta-Galactosidase was produced. From 51 compounds tested 21 were used as single sources of carbon. The major respiratory lipoquinone was ubiquinone-10. The predominant cellular fatty acids were 16 : 0, 18 : 1omega7c and 16 : 1omega7c (from summed feature 3). Hydroxy fatty acids 14 : 0 2-OH and 15 : 0 iso 2-OH were present as well (from summed feature 4). The polar lipids detected in strain FA2(T) were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, sphingoglycolipid and two unidentified phospholipids. The DNA G+C content of strain FA2(T) was 61 mol%. DNA-DNA hybridization experiments showed 27.6 % relatedness between strain FA2(T) and S. adhaesiva DSM 7418(T). Based upon phenotypic and molecular evidence, a novel species of the genus Sphingomonas is proposed, Sphingomonas phyllosphaerae sp. nov., with strain FA2(T) (=LMG 21958(T)=CECT 5832(T)) as the type strain. PMID- 15545450 TI - Nocardioides aestuarii sp. nov., isolated from tidal flat sediment. AB - A strictly aerobic, non-motile, rod-shaped actinomycete, designated strain JC2056(T), was isolated from a sediment sample of getbol, the tidal flat of Korea. Results of 16S rRNA gene sequence analysis indicated that the isolate belonged to the genus Nocardioides, with the highest similarity being to Nocardioides luteus KCTC 9575(T) (95.7 %). The major menaquinone was MK-8(H(4)), and predominant cellular fatty acids were iso-16 : 0 and iso-16 : 1 H. The DNA G+C content was 70 mol%. Based on the morphological, physiological, biochemical and chemotaxonomical data presented in this study, strain JC2056(T) should be classified as a novel species, for which the name Nocardioides aestuarii sp. nov. is proposed; the type strain is JC2056(T) (=IMSNU 14029(T)=KCTC 9921(T)=JCM 12125(T)). PMID- 15545451 TI - Burkholderia tropica sp. nov., a novel nitrogen-fixing, plant-associated bacterium. AB - In an ecological survey of nitrogen-fixing bacteria isolated from the rhizosphere and as endophytes of sugarcane, maize and teosinte plants in Brazil, Mexico and South Africa, a new phylogenetically homogeneous group of N(2)-fixing bacteria was identified within the genus Burkholderia. This polyphasic taxonomic study included microscopic and colony morphology, API 20NE tests and growth on different culture media at different pH and temperatures, as well as carbon source assimilation tests and whole-cell protein pattern analysis. Analysis of 16S rRNA gene sequences showed 99.2-99.9 % similarity within the novel species and 97.2 % similarity to the closest related species, Burkholderia sacchari. The novel species was composed of four distinct amplified 16S rDNA restriction analysis groups. The DNA-DNA reassociation values within the novel species were greater than 70 % and less than 42 % for the closest related species, B. sacchari. Based on these results and on many phenotypic characteristics, a novel N(2)-fixing species is proposed for the genus Burkholderia, Burkholderia tropica sp. nov., with the type strain Ppe8(T) (=ATCC BAA-831(T)=LMG 22274(T)=DSM 15359(T)). B. tropica was isolated from plants grown in geographical regions with climates ranging from temperate subhumid to hot humid. PMID- 15545452 TI - Transfer of Bacillus halodenitrificans Denariaz et al. 1989 to the genus Virgibacillus as Virgibacillus halodenitrificans comb. nov. AB - A Gram-variable, endospore-forming moderately halophilic rod, strain SF-121, was isolated from a marine solar saltern of the Yellow Sea in Korea. The result of 16S rRNA gene sequence analysis showed that strain SF-121 has highest sequence similarity (99.7 %) with the type strain of Bacillus halodenitrificans. Phylogenetic analyses based on 16S rRNA gene sequences revealed that B. halodenitrificans DSM 10037(T) and strain SF-121 are more closely related to the genus Virgibacillus than to the genus Bacillus. Strain SF-121 and B. halodenitrificans DSM 10037(T) exhibited 16S rRNA gene similarity levels of 95.3 97.5 % with the type strains of Virgibacillus species and 94.0 % with the type strain of Bacillus subtilis. DNA-DNA relatedness and phenotypic data indicated that B. halodenitrificans DSM 10037(T) and strain SF-121 are members of the same species. B. halodenitrificans DSM 10037(T) and strain SF-121 exhibited DNA-DNA relatedness values of 9-11 % with the type strains of Virgibacillus carmonensis and Virgibacillus marismortui. On the basis of the phenotypic, chemotaxonomic, phylogenetic and genetic data, B. halodenitrificans should be reclassified in the genus Virgibacillus as Virgibacillus halodenitrificans comb. nov. PMID- 15545453 TI - Mahella australiensis gen. nov., sp. nov., a moderately thermophilic anaerobic bacterium isolated from an Australian oil well. AB - A novel Gram-positive, anaerobic and moderately thermophilic bacterium, strain 50 1 BON(T), was isolated from an Australian terrestrial oil reservoir. Cells were spore-forming straight rods, motile by peritrichous flagella. The optimum growth conditions were 50 degrees C, pH 7.5 and 0.1 % NaCl. Strain 50-1 BON(T) fermented arabinose, cellobiose, fructose, galactose, glucose, mannose, sucrose, xylose and yeast extract. Glucose was fermented mainly into lactate, formate, hydrogen and CO(2). The major end product of pyruvate fermentation was acetate together with H(2) and CO(2). Thiosulfate, sulfate, elemental sulfur and nitrate were not used as terminal electron acceptors. The DNA G+C content was 55.5 mol%. The closest phylogenetic relative of strain 50-1 BON(T) was Thermoanaerobacterium thermosulfurigenes (16S rRNA gene sequence similarity of 85.7 %). As strain 50-1 BON(T) was physiologically and phylogenetically different from members of the order 'Thermoanaerobacteriales', it is proposed that strain 50-1 BON(T) (=DSM 15567(T)=CIP 107919(T)) be classified as the type strain of a novel species of a new genus, Mahella australiensis gen. nov., sp. nov. PMID- 15545454 TI - Enterococcus saccharominimus sp. nov., from dairy products. AB - Four isolates, which were obtained from Belgian, Moroccan and Romanian dairy products, constituted a homogeneous but unidentified taxon after screening with whole-cell protein fingerprinting. Complete 16S rRNA gene sequence analysis classified representative strains in the genus Enterococcus. Highest sequence similarities of 98.6 and 98.0 % were obtained with the species Enterococcus sulfureus and Enterococcus saccharolyticus, respectively. Growth characteristics, biochemical features, tRNA intergenic length polymorphism analysis, DNA-DNA hybridization and DNA G+C contents of selected strains demonstrated that they represent a single, novel Enterococcus species. It differs phenotypically from other enterococci in characteristics commonly considered as typical of this genus: no growth in 6.5 % NaCl or 0.4 % sodium azide, and no acid production from a wide range of carbohydrates. The name Enterococcus saccharominimus sp. nov. is proposed for this novel species; the type strain (LMG 21727(T)=CCM 7220(T)) was isolated from contaminated pasteurized cow's milk. PMID- 15545455 TI - Jonesia quinghaiensis sp. nov., a new member of the suborder Micrococcineae. AB - A coryneform strain isolated from soda lake mud in China corresponded in chemotaxonomic characteristics such as peptidoglycan type A4alpha l-lys-l-ser-d Glu and major menaquinone MK-9, as well as in its DNA base composition (57 mol% G+C), to its phylogenetic neighbour Jonesia denitrificans. Differences in phenotypic characteristics and the phylogenetic distance (96.6 % 16S rRNA gene sequence similarity) from J. denitrificans justify the proposal of a second species of the genus Jonesia, Jonesia quinghaiensis sp. nov., with the type strain QH3A7(T) (=DSM 15701(T)=CGMCC 1.3459(T)). PMID- 15545456 TI - Sphingomonas oligophenolica sp. nov., a halo- and organo-sensitive oligotrophic bacterium from paddy soil that degrades phenolic acids at low concentrations. AB - The taxonomic position of a halo- and organo-sensitive, oligotrophic soil bacterium, strain S213(T), was investigated. Cells were Gram-negative, non motile, strictly aerobic, yellow-pigmented rods of short to medium length on diluted nutrient broth. When 0.1-0.4 % (w/v) NaCl was added to diluted media composed of peptone and meat extract, growth was inhibited with increasing NaCl concentration and the cells became long aberrant forms. When 6 mM CaCl(2) was added, the cells grew quite normally and aberrant cells were no longer found at 0.1-0.5 % (w/v) NaCl. Chemotaxonomically, strain S213(T) contains chemical markers that indicate its assignment to the Sphingomonadaceae: the presence of ubiquinone Q-10 as the predominant respiratory quinone, C(18 : 1) and C(16 : 0) as major fatty acids, C(14 : 0) 2-OH as the major 2-hydroxy fatty acid and sphingoglycolipids. 16S rRNA gene sequence analysis indicated that strain S213(T) belongs to the genus Sphingomonas, exhibiting high sequence similarity to the 16S rRNA gene sequences of Sphingomonas mali IFO 15500(T) (98.3 %), Sphingomonas pruni IFO 15498(T) (98.0 %), Sphingomonas asaccharolytica IFO 15499(T) (97.9 %) and Sphingomonas echinoides DSM 1805(T) (97.8 %). The results of DNA-DNA hybridization experiments and its phenotypic characteristics clearly distinguished the strain from its nearest neighbours and demonstrate that strain S213(T) represents a novel Sphingomonas species, for which the name Sphingomonas oligophenolica sp. nov. is proposed. The type strain is S213(T) (=JCM 12082(T)=CIP 107926(T)). PMID- 15545457 TI - Corynebacterium ciconiae sp. nov., isolated from the trachea of black storks (Ciconia nigra). AB - Eight unidentified Gram-positive, rod-shaped organisms were recovered from the tracheas of apparently healthy black storks (Ciconia nigra) and subjected to a polyphasic taxonomic analysis. Based on cellular morphology and biochemical criteria the isolates were tentatively assigned to the genus Corynebacterium, although three of the organisms did not appear to correspond to any recognized species. Comparative 16S rRNA gene sequencing studies demonstrated that all of the isolates were phylogenetically members of the genus Corynebacterium. Five strains were genotypically identified as representing Corynebacterium falsenii, whereas the remaining three strains represented a hitherto unknown subline, associated with a small subcluster of species that includes Corynebacterium mastitidis and its close relatives. On the basis of phenotypic and phylogenetic evidence, it is proposed that the unknown isolates from black storks represent a novel species within the genus Corynebacterium, for which the Corynebacterium ciconiae sp. nov. is proposed. The type strain is CECT 5779(T) (=BS13(T)=CCUG 47525(T)). PMID- 15545458 TI - Geobacillus debilis sp. nov., a novel obligately thermophilic bacterium isolated from a cool soil environment, and reassignment of Bacillus pallidus to Geobacillus pallidus comb. nov. AB - Several aerobic, motile, rod-shaped, thermophilic, spore-forming Geobacillus bacteria predominantly giving a Gram-positive staining reaction were isolated from a cool soil environment in Northern Ireland and taxonomically investigated. Two isolates, F10 and Tf(T), showed low 16S rRNA gene sequence similarity to recognized members of the genus Geobacillus. Phylogenetic tree investigation using neighbour-joining, maximum-likelihood and parsimony methods indicated that strains F10 and Tf(T) represent a single novel species, for which the name Geobacillus debilis sp. nov. is proposed, with type strain Tf(T) (=DSM 16016(T)=NCIMB 13995(T)) and which belongs to a subgroup of the genus Geobacillus comprising Geobacillus toebii and Geobacillus caldoxylosilyticus. However, G. debilis showed closest affinities to Bacillus pallidus, which we propose should become Geobacillus pallidus comb. nov. PMID- 15545459 TI - Description of 'Candidatus Helicobacter heilmannii' based on DNA sequence analysis of 16S rRNA and urease genes. AB - While Helicobacter pylori is accepted as the major bacterial agent of gastric disease in humans, some patients and many animals are infected with a larger, tightly helical-shaped bacterium previously referred to as 'Helicobacter heilmannii' or 'Gastrospirillum hominis'. Taxonomic classification of these bacteria has been hampered by the inability to cultivate them in vitro and by the inadequate discriminatory power of 16S rRNA gene sequence analysis. This study describes the detection and phylogenetic analysis of 26 different gastrospirillum isolates from humans and animals, which incorporates sequence data based on the 16S rRNA and urease genes. Fifteen gastrospirilla detected in humans, primates and pigs clustered with 'Candidatus Helicobacter suis', thus expanding the host range for this organism. By comparison, based on 16S rRNA data, the remaining 11 gastrospirilla could not be differentiated from Helicobacter felis, Helicobacter bizzozeronii and Helicobacter salomonis. However, urease gene sequence analysis allowed for the discrimination of this latter group into four discrete clusters, three of which contained the above recognized species. The fourth cluster contained isolates from human and feline hosts, and should provisionally be considered a unique bacterial species, for which the name 'Candidatus Helicobacter heilmannii' is proposed. PMID- 15545460 TI - Reclassification of Staphylococcus pulvereri Zakrzewska-Czerwinska et al. 1995 as a later synonym of Staphylococcus vitulinus Webster et al. 1994. AB - A polyphasic taxonomic approach was applied to strains of the species Staphylococcus vitulinus and Staphylococcus pulvereri in order to clarify their taxonomic relatedness. Four reference strains, representing both species, and seven strains isolated from human clinical material were characterized by biotyping, ribotyping and SDS-PAGE analysis of whole-cell proteins, and none of the screening approaches allowed the two taxa to be distinguished. DNA-DNA hybridization experiments between four selected representative strains, including the type strains, confirmed that Staphylococcus pulvereri is a later synonym of Staphylococcus vitulinus. PMID- 15545461 TI - Erwinia toletana sp. nov., associated with Pseudomonas savastanoi-induced tree knots. AB - Gram-negative bacteria were isolated from knots induced by Pseudomonas savastanoi in olive trees (Olea europaea L.). A total of nine endophytic bacterial strains were isolated, each from inside a different tree knot. Biochemical characterization indicated that all the strains belong to the family Enterobacteriaceae. Phylogenetic analyses of the 16S rRNA genes of these novel isolates revealed that they formed a homogeneous cluster within Erwinia species. DNA signatures of these isolates were identical to those described for the genus Erwinia. The strains formed a homogeneous group as shown by DNA-DNA hybridization analysis and numerical analysis of phenotypic data, clearly differentiated from all species of Erwinia with validly published names. The data provide strong evidence of the differentiation of these strains from the most closely related species. Therefore, these isolates represent a novel species, for which the name Erwinia toletana sp. nov. is proposed. The isolates are available at CFBP, CECT and ATCC. The G+C content is 52+/-0.5 mol%. The type strain is CFBP 6631(T) (=A37(T)=ATCC 700880(T)=CECT 5263(T)). PMID- 15545462 TI - Proposals of Curvibacter gracilis gen. nov., sp. nov. and Herbaspirillum putei sp. nov. for bacterial strains isolated from well water and reclassification of [Pseudomonas] huttiensis, [Pseudomonas] lanceolata, [Aquaspirillum] delicatum and [Aquaspirillum] autotrophicum as Herbaspirillum huttiense comb. nov., Curvibacter lanceolatus comb. nov., Curvibacter delicatus comb. nov. and Herbaspirillum autotrophicum comb. nov. AB - Two strains of curved bacteria, 7-1(T) and 7-2(T), isolated from well water, were phylogenetically examined to determine their taxonomic position. Strain 7-1(T) is a Gram-negative, slightly curved rod. Analysis of the 16S rRNA gene sequence showed that strain 7-1(T) formed a cluster with [Aquaspirillum] delicatum and [Pseudomonas] lanceolata. It has some similar characteristics to [A.] delicatum and [P.] lanceolata, but has sufficient distance to separate it from other genera. DNA-DNA hybridization analysis, as well as chemotaxonomic and morphological studies, demonstrated that strain 7-1(T), [A.] delicatum and [P.] lanceolata belong to a new genus, Curvibacter gen. nov. Strain 7-1(T) (=IAM 15033(T)=ATCC BAA-807(T)) is classified as the type strain of Curvibacter gracilis gen. nov., sp. nov., and [A.] delicatum and [P.] lanceolata are classified as Curvibacter delicatus comb. nov. and Curvibacter lanceolatus comb. nov., respectively. Strain 7-2(T) is a Gram-negative spirillum. Phylogenetic study based on the 16S rRNA gene sequences showed that it formed a cluster with the members of the genus Herbaspirillum, [Pseudomonas] huttiensis and [Aquaspirillum] autotrophicum. The classification is therefore proposed of strain 7-2(T) (=IAM 15032(T)=ATCC BAA-806(T)) as the type strain of Herbaspirillum putei sp. nov., and [P.] huttiensis and [A.] autotrophicum are transferred to the genus Herbaspirillum as Herbaspirillum huttiense comb. nov. and Herbaspirillum autotrophicum comb. nov., respectively. PMID- 15545463 TI - Porphyrobacter donghaensis sp. nov., isolated from sea water of the East Sea in Korea. AB - Two Gram-negative, motile, non-spore-forming, bacteriochlorophyll a-containing slightly halophilic strains, SW-132(T) and SW-158, were isolated from sea water of the East Sea in Korea, and subjected to a polyphasic taxonomic study. The two isolates were characterized chemotaxonomically as having Q-10 as the predominant respiratory lipoquinone and major amounts of unsaturated fatty acids C(18 : 1)omega7c and C(17 : 1)omega6c. The DNA G+C contents of the two strains were in the range 66.8-65.9 mol%. The 16S rRNA gene sequences of strains SW-132(T) and SW 158 were 99.9 % (1 nt difference) similar and their mean level of DNA-DNA relatedness was 86 %. The 16S rRNA gene sequence analysis showed that strains SW 132(T) and SW-158 are phylogenetically closely related to Porphyrobacter species and Erythromicrobium ramosum. Similarities between the 16S rRNA gene sequences of strains SW-132(T) and SW-158 and the type strains of Porphyrobacter species and E. ramosum ranged from 97.8 to 99.0 %. DNA-DNA relatedness data indicated that strains SW-132(T) and SW-158 are members of a genomic species that is separate from the four Porphyrobacter species. On the basis of phenotypic and phylogenetic data and genetic distinctiveness, strains SW-132(T) (=KCTC 12229(T)=DSM 16220(T)) and SW-158 (=KCTC 12230) are classified as a novel Porphyrobacter species, for which the name Porphyrobacter donghaensis sp. nov. is proposed. PMID- 15545464 TI - 'Candidatus Burkholderia calva' and 'Candidatus Burkholderia nigropunctata' as leaf gall endosymbionts of African Psychotria. AB - Phylogenetic 16S rRNA gene analysis was used to assign the bacterial leaf nodulating endosymbionts of two tropical African Psychotria species to the genus Burkholderia. The microsymbionts of the different Psychotria hosts were recognized as distinct and novel species of Burkholderia on the basis of relatively low intersequence similarities and sufficiently large evolutionary distances when compared with each other and their closest validly named neighbours. The obligate endosymbiotic nature of the bacteria prevented their in vitro cultivation and the deposition of type strains to culture collections. Therefore, the provisional status Candidatus is assigned to the bacterial partners of Psychotria calva and Psychotria nigropunctata, with the proposal of the names 'Candidatus Burkholderia calva' and 'Candidatus Burkholderia nigropunctata', respectively. PMID- 15545465 TI - Bacillus macyae sp. nov., an arsenate-respiring bacterium isolated from an Australian gold mine. AB - A strictly anaerobic arsenate-respiring bacterium isolated from a gold mine in Bendigo, Victoria, Australia, belonging to the genus Bacillus is described. Cells are Gram-positive, motile rods capable of respiring with arsenate and nitrate as terminal electron acceptors using a variety of substrates, including acetate as the electron donor. Reduction of arsenate to arsenite is catalysed by a membrane bound arsenate reductase that displays activity over a broad pH range. Synthesis of the enzyme is regulated; maximal activity is obtained when the organism is grown with arsenate as the terminal electron acceptor and no activity is detectable when it is grown with nitrate. Mass of the catalytic subunit was determined to be approximately 87 kDa based on ingel activity stains. The closest phylogenetic relative, based on 16S rRNA gene sequence analysis, is Bacillus arseniciselenatis, but DNA-DNA hybridization experiments clearly show that strain JMM-4(T) represents a novel Bacillus species, for which the name Bacillus macyae sp. nov. is proposed. The type strain is JMM-4(T) (=DSM 16346(T)=JCM 12340(T)). PMID- 15545466 TI - Pseudoxanthomonas mexicana sp. nov. and Pseudoxanthomonas japonensis sp. nov., isolated from diverse environments, and emended descriptions of the genus Pseudoxanthomonas Finkmann et al. 2000 and of its type species. AB - Three mesophilic bacteria (strains AMX 26B(T), UR374_02 and 12-3(T)) isolated respectively from an anaerobic digester, human urine and urban riverside soil were characterized. Cells were Gram-negative, motile, non-sporulating, straight to curved rods with one polar flagellum and had a strictly respiratory metabolism with O(2) as the preferential terminal electron acceptor. Phylogenetic analysis based on 16S rRNA gene sequences revealed that all strains clustered within the Xanthomonadaceae branch of the Proteobacteria. Isolates AMX 26B(T) and UR374_02 exhibited 100 % 16S rRNA gene sequence similarity and both were related to strain 12-3(T) (99.6 % similarity). The closest relative of all the isolates was Pseudoxanthomonas broegbernensis DSM 12573(T) (similarity 97.1-97.5 %), and they were equidistantly related to Xanthomonas species (95.4-96.6 %), Stenotrophomonas species (95.3-96.1 %) and Pseudoxanthomonas taiwanensis ATCC BAA-4040(T) (95.3 95.4 %). Chemotaxonomic and biochemical data (branched-chain cellular fatty acid pattern without C(13 : 0) iso 3-OH, ubiquinone with eight isoprenoid units, limited range of substrates used, ability to reduce nitrite but not nitrate with the production of N(2)O) supported their affiliation to the genus Pseudoxanthomonas. The results of DNA-DNA hybridization and/or phenotypic analysis allowed them to be differentiated from the two Pseudoxanthomonas species with validly published names and showed that strain 12-3(T) was genomically and phenotypically distinct from the other two isolates. On the basis of these results, two novel species of the genus Pseudoxanthomonas are proposed: Pseudoxanthomonas mexicana sp. nov., consisting of strains AMX 26B(T) (=ATCC 700993(T)=CIP 106674(T)=JCM 11524(T)) (type strain) and UR374_02 (=DSM 15133), and Pseudoxanthomonas japonensis sp. nov., consisting of strain 12-3(T) (=CCUG 48231(T)=CIP 107388(T)=JCM 11525(T)). The report of these two novel species leads to the emendation of the description of the genus Pseudoxanthomonas and the re evaluation of the phenotype of P. broegbernensis DSM 12573(T) necessitates the emendation of its description. PMID- 15545467 TI - Acetanaerobacterium elongatum gen. nov., sp. nov., from paper mill waste water. AB - Two mesophilic anaerobic bacterial strains (Z7(T) and Z1) were isolated from waste water sludge of the Xinanzhang paper mill, Beijing, China. The strains were Gram-positive, non-spore-forming and motile. Cells were thin rods (0.2-0.4x4.0 8.0 microm). Growth of the strains was observed at 20-42 degrees C and pH 5.0 7.5. Both strains hydrolysed gelatin and aesculin and fermented several kinds of mono-, di- and oligosaccharides. The fermentation end products formed from glucose were acetate, ethanol, hydrogen and carbon dioxide. The predominant cellular fatty acids were the branched-chain fatty acids isoC(15 : 0) (42.83 %) and isoC(14 : 0) (32.11 %). The DNA G+C contents of strains Z7(T) and Z1 were 50.4 and 48.6 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the isolates represent a new phyletic sublineage within the Clostridium leptum rRNA cluster, with <91 % 16S rRNA gene sequence similarity to currently described species. On the basis of polyphasic evidence from this study, Acetanaerobacterium elongatum gen. nov., sp. nov., a novel genus and species, is proposed, with strain Z7(T) (=JCM 12359(T)=AS 1.5012(T)) as the type strain. PMID- 15545468 TI - Saccharibacter floricola gen. nov., sp. nov., a novel osmophilic acetic acid bacterium isolated from pollen. AB - Three Gram-negative, aerobic, rod-shaped bacterial strains were isolated, from the pollen of Japanese flowers, as producers of xylitol; these strains were subjected to a polyphasic taxonomic study. Phylogenetic analyses of the 16S rRNA gene sequences demonstrated that these three isolates formed a new cluster within a group of acetic acid bacteria in the alpha-Proteobacteria. The characteristics of the three isolates were as follows: (i) their predominant quinone was Q-10; (ii) their cellular fatty acid profile contained major amounts of 2-hydroxy acids and an unsaturated straight-chain acid (C(18 : 1)omega7c); and (iii) their DNA G+C contents were in the range 51.9-52.3 mol%, which is around the lower limit of the reported range for the genera of acetic acid bacteria. The negligible or very weak productivity of acetic acid from ethanol and the osmophilic growth properties distinguished these strains from other acetic acid bacteria. The unique phylogenetic and phenotypic characteristics suggest that the three isolates should be classified within a novel genus and species with the proposed name Saccharibacter floricola gen. nov., sp. nov. The type strain is strain S 877(T) (=AJ 13480(T)=JCM 12116(T)=DSM 15669(T)). PMID- 15545469 TI - Methylobacterium nodulans sp. nov., for a group of aerobic, facultatively methylotrophic, legume root-nodule-forming and nitrogen-fixing bacteria. AB - Data on 72 non-pigmented bacterial strains that specifically induce nitrogen fixing root nodules on the legume species Crotalaria glaucoides, Crotalaria perrottetii and Crotalaria podocarpa are reviewed. By SDS-PAGE analysis of total protein patterns and by 16S rRNA PCR-RFLP, these strains form a homogeneous group that is separate from other legume root-nodule-forming bacteria. The 16S rRNA gene-based phylogeny indicates that these bacteria belong to the genus Methylobacterium. They can grow on C(1) compounds such as methanol, formate and formaldehyde but not methylamine as sole carbon source, and carry an mxaF gene, encoding methanol dehydrogenase, which supports their methylotrophic metabolism. Presence of a nodA nodulation gene, and ability to nodulate plants of Crotalaria species and to fix nitrogen are features that separate the strains currently included in this group from other members of the genus Methylobacterium. The present study includes additional genotypic and phenotypic characterization of this novel Methylobacterium species, i.e. nifH gene sequence, morphology, physiology, enzymic and carbon source assimilation tests and antibiotic resistance. The name Methylobacterium nodulans sp. nov. (type strain, ORS 2060(T)=CNCM I 2342(T)=LMG 21967(T)) is proposed for this group of root-nodule forming bacteria. PMID- 15545470 TI - Haloferax sulfurifontis sp. nov., a halophilic archaeon isolated from a sulfide- and sulfur-rich spring. AB - A pleomorphic, extremely halophilic archaeon (strain M6(T)) was isolated from a sulfide- and sulfur-rich spring in south-western Oklahoma (USA). It formed small (0.8-1.0 mm), salmon pink, elevated colonies on agar medium. The strain grew in a wide range of NaCl concentrations (6 % to saturation) and required at least 1 mM Mg(2+) for growth. Strain M6(T) was able to reduce sulfur to sulfide anaerobically. 16S rRNA gene sequence analysis indicated that strain M6(T) belongs to the family Halobacteriaceae, genus Haloferax; it showed 96.7-98.0 % similarity to other members of the genus with validly published names and 89 % similarity to Halogeometricum borinquense, its closest relative outside the genus Haloferax. Polar lipid analysis and DNA G+C content further supported placement of strain M6(T) in the genus Haloferax. DNA-DNA hybridization values, as well as biochemical and physiological characterization, allowed strain M6(T) to be differentiated from other members of the genus Haloferax. A novel species, Haloferax sulfurifontis sp. nov., is therefore proposed to accommodate the strain. The type strain is M6(T) (=JCM 12327(T)=CCM 7217(T)=DSM 16227(T)=CIP 108334(T)). PMID- 15545471 TI - Loktanella hongkongensis sp. nov., a novel member of the alpha-Proteobacteria originating from marine biofilms in Hong Kong waters. AB - A Gram-negative, non-motile, non-spore-forming, short rod-shaped bacterium (UST950701-009P(T)) was isolated from a marine biofilm in Hong Kong waters. Colonies are pink in colour, convex with a smooth surface and entire edge. Brown diffusible pigment is produced. Whitish colonies, with otherwise identical morphology, emerge from every culture upon ageing. The white colonies can be maintained as separate cultures (UST950701-009W) without turning pink. UST950701 009P(T) and UST950701-009W have identical 16S rRNA gene sequences and similar G+C (65.9-66.2 mol%) and fatty acid (86.22-88.52 % 18 : 1omega7c) contents. Phylogenetic analysis of the 16S rRNA gene sequence places UST950701-009P(T) within the Rhodobacter group of the alpha-subclass of the Proteobacteria. The nearest neighbours belong to the genus Loktanella, with similarity values ranging from 94.5 to 95.5 %. Data on G+C and fatty acid contents support the affiliation to the genus Loktanella. UST950701-009P(T) and -009W are heterotrophic, strictly aerobic and require NaCl for growth (2.0-14.0 %). Both grow in pH 5.0-10.0 and at 8-44 degrees C. Both are positive in oxidase, catalase and beta-galactosidase tests, but they differ in the pattern of carbohydrate oxidation and assimilation. Molecular evidence together with phenotypic characteristics shows that UST950701 009P(T) constitutes a novel species within the genus Loktanella. The name Loktanella hongkongensis sp. nov. is proposed; the type strain is UST950701 009P(T) (=NRRL B-41039(T)=JCM 12479(T)) and a morphovar is UST950701-009W (=NRRL B-41040=JCM 12480). PMID- 15545472 TI - Taxonomy of the genus Cupriavidus: a tale of lost and found. AB - DNA-DNA hybridization experiments and an evaluation of phenotypic characteristics, DNA base ratios and 16S rRNA gene sequences demonstrated that Wautersia eutropha (Davies 1969) Vaneechoutte et al. 2004, the type species of the genus Wautersia, is a later synonym of Cupriavidus necator Makkar and Casida 1987, the type species of the genus Cupriavidus. In conformity with Rules 15, 17, 23a and 37a(1) of the International Code of Nomenclature of Bacteria, the genus name Cupriavidus has priority over the genus name Wautersia, and all other members of the genus Wautersia are reclassified into Cupriavidus as Cupriavidus basilensis comb. nov. (type strain LMG 18990(T)=DSM 11853(T)), Cupriavidus campinensis comb. nov. (type strain LMG 19282(T)=CCUG 44526(T)), Cupriavidus gilardii comb. nov. (type strain LMG 5886(T)=CCUG 38401(T)), Cupriavidus metallidurans comb. nov. (type strain LMG 1195(T)=DSM 2839(T)), Cupriavidus oxalaticus comb. nov. (type strain LMG 2235(T)=CCUG 2086(T)=DSM 1105(T)), Cupriavidus pauculus comb. nov. (type strain LMG 3244(T)=CCUG 12507(T)), Cupriavidus respiraculi comb. nov. (type strain LMG 21510(T)=CCUG 46809(T)) and Cupriavidus taiwanensis comb. nov. (type strain LMG 19424(T)=CCUG 44338(T)). PMID- 15545473 TI - Streptococcus equi subsp. ruminatorum subsp. nov., isolated from mastitis in small ruminants. AB - Six isolates of an unknown Gram-positive, catalase-negative, chain-forming, coccus-shaped organism isolated from ovine and caprine mastitis were characterized by phenotypic and molecular taxonomic methods. On the basis of cellular morphology and the results of biochemical tests, the organism was tentatively identified as a streptococcal species. Comparative 16S rRNA gene sequencing studies confirmed that the organism is a member of the genus Streptococcus, with Streptococcus equi as its closest phylogenetic relative (98.8 % similarity). DNA-DNA pairing studies showed that the unidentified organism displayed more than 70 % relatedness to the type strains of S. equi subsp. equi and subsp. zooepidemicus. Despite the relatively high DNA-DNA reassociation values, biotyping and ribotyping allowed clear differentiation of the unknown bacterium from the two recognized subspecies of S. equi. On the basis of phenotypic and molecular genetic evidence, it is proposed that the unknown Streptococcus isolates from ovine and caprine mastitis be classified as a novel subspecies, Streptococcus equi subsp. ruminatorum subsp. nov. The type strain is CECT 5772(T) (=CCUG 47520(T)=Mt 167(T)). PMID- 15545474 TI - Sulfuricurvum kujiense gen. nov., sp. nov., a facultatively anaerobic, chemolithoautotrophic, sulfur-oxidizing bacterium isolated from an underground crude-oil storage cavity. AB - A facultatively anaerobic, chemolithoautotrophic, sulfur-oxidizing bacterium, strain YK-1(T), was isolated from an underground crude-oil storage cavity at Kuji in Iwate, Japan. The cells were motile, curved rods and had a single polar flagellum. Optimum growth occurred in a low-strength salt medium at pH 7.0 and 25 degrees C. It utilized sulfide, elemental sulfur, thiosulfate and hydrogen as the electron donors and nitrate as the electron acceptor under anaerobic conditions, but it did not use nitrite. Oxygen also served as the electron acceptor under the microaerobic condition (O(2) in the head space 1 %). It did not grow on sugars, organic acids or hydrocarbons as carbon and energy sources. The DNA G+C content of strain YK-1(T) was 45 mol%. Phylogenetic analysis, based on the 16S rRNA gene sequence, showed that its closest relative was Thiomicrospira denitrificans in the 'Epsilonproteobacteria', albeit with low homology (90 %). On the basis of physiological and phylogenetic data, strain YK-1(T) should be classified into a novel genus and species, for which the name Sulfuricurvum kujiense gen. nov., sp. nov. is proposed. The type strain is YK-1(T) (=JCM 11577(T)=MBIC 06352(T)=ATCC BAA-921(T)). PMID- 15545475 TI - Nocardia xishanensis sp. nov., a novel actinomycete isolated from soil. AB - The taxonomic position of a soil isolate, strain 276(T), was established using a polyphasic approach. The organism showed a range of chemical and morphological properties consistent with its classification in the genus Nocardia. An almost complete 16S rRNA gene sequence determined for the strain was aligned with corresponding sequences of representatives of the genus Nocardia and related taxa using three tree-making algorithms. The organism formed a distinct phyletic line within the evolutionary radiation occupied by the genus Nocardia and was most closely related to the type strain of Nocardia abscessus. However, the two strains shared a low DNA-DNA relatedness value and were readily distinguished using a combination of phenotypic properties. The combined genotypic and phenotypic data show that strain 276(T) should be assigned to the genus Nocardia as a novel species. The name proposed for this new taxon is Nocardia xishanensis sp. nov. The type strain is 276(T) (=CGMCC 4.1860(T)=JCM 12160(T)). PMID- 15545476 TI - Pseudovibrio denitrificans gen. nov., sp. nov., a marine, facultatively anaerobic, fermentative bacterium capable of denitrification. AB - Two denitrifying strains of heterotrophic, facultatively anaerobic, marine bacteria, designated DN34(T) and DN33, were isolated from sea-water samples collected in Nanwan Bay, Kenting National Park, Taiwan. They were Gram-negative. Cells in late exponential to early stationary phase of growth were predominantly straight or curved rods, but Y- or V-shaped forms were also observed. They were motile by means of one to several lateral or subpolar flagella. Both strains required NaCl for growth and exhibited optimal growth at about 30 degrees C, pH 8 and 3 % NaCl. They were capable of anaerobic growth by carrying out denitrifying metabolism using nitrate, nitrite or nitrous oxide as terminal electron acceptors or, alternatively, by fermenting glucose, mannose, sucrose or trehalose as substrates. Anaerobic fermentative growth on glucose resulted in formation of various organic acids, including formate, lactate, acetate, pyruvate and fumarate. The major cellular fatty acids were 2-OH-14 : 0, 3-OH-14 : 0 and 16 : 0. DN34(T) and DN33 had DNA G+C contents of 51.7 and 51.6 mol%, respectively. Physiological characterization, together with phylogenetic analysis based on 16S rRNA gene sequence analysis, revealed that the two denitrifying strains could be accommodated in a novel genus, for which the name Pseudovibrio gen. nov. is proposed. Pseudovibrio denitrificans sp. nov. is the type species, with DN34(T) (=BCRC 17323(T)=JCM 12308(T)) as the type strain. PMID- 15545477 TI - Mycobacterium pyrenivorans sp. nov., a novel polycyclic-aromatic-hydrocarbon degrading species. AB - The taxonomic position of a polycyclic-aromatic-hydrocarbon-degrading bacterium, strain 17A3(T), isolated from contaminated soil was determined using a combination of phenotypic and genotypic properties. The isolate showed phenotypic properties that were diagnostic for species of the genus Mycobacterium. Comparative 16S rRNA gene sequence analysis assigned 17A3(T) to the 16S rRNA gene subgroup that contains Mycobacterium aurum, Mycobacterium austroafricanum, Mycobacterium vaccae and Mycobacterium vanbaalenii, but it could clearly be distinguished from these species using a combination of physiological, chemotaxonomic markers and internal rRNA gene spacer analyses. The data showed that strain 17A3(T) (=DSM 44605(T)=NRRL B-24244(T)) merits recognition as the type strain of a novel species of the genus Mycobacterium. The name Mycobacterium pyrenivorans sp. nov. is proposed for the species because of its ability to use pyrene as a sole source of carbon and energy. PMID- 15545478 TI - Kaistella koreensis gen. nov., sp. nov., a novel member of the Chryseobacterium Bergeyella-Riemerella branch. AB - Gram-negative, non-spore-forming, rod-shaped, yellow-pigmented bacteria isolated from a freshwater stream in Korea were investigated to determine their taxonomic position. Complete 16S rRNA gene sequence analysis indicated that the organisms should be placed in the Chryseobacterium-Bergeyella-Riemerella branch in the family Flavobacteriaceae. Phylogenetically, the strains were most closely related to Chryseobacterium balustinum ATCC 33487(T) and Chryseobacterium scophthalmum LMG 13028(T) (94.3 and 94.1 % 16S rRNA gene sequence similarity, respectively) and they clustered on a separate well-supported branch. The strains contained menaquinone MK-6 as the predominant respiratory quinone and showed higher G+C contents (41.7 mol%) than other species in the Chryseobacterium-Bergeyella Riemerella branch and i-C(15 : 0) as a major fatty acid (47-52 %). The phylogenetic distances from any species with validly published names and their phenotypic properties confirmed that the strains constitute a separate species in a new genus, for which the name Kaistella koreensis gen. nov., sp. nov. is proposed (type strain Chj707(T)=KCTC 12107(T)=IAM 15050(T)). PMID- 15545479 TI - Thiomicrospira thermophila sp. nov., a novel microaerobic, thermotolerant, sulfur oxidizing chemolithomixotroph isolated from a deep-sea hydrothermal fumarole in the TOTO caldera, Mariana Arc, Western Pacific. AB - A novel thermotolerant bacterium, designated strain I78(T), was isolated from a self-temperature-recording in situ colonization system deployed in a hydrothermal diffusing flow (maximal temperature 78 degrees C) at the TOTO caldera in the Mariana Arc, Western Pacific. Cells were highly motile curved rods with a single polar flagellum. Growth was observed at 15-55 degrees C (optimum 35-40 degrees C; 60 min doubling time) and pH 5.0-8.0 (optimum pH 6.0). The isolate was a microaerobic chemolithomixotroph capable of using thiosulfate, elemental sulfur or sulfide as the sole energy source, and molecular oxygen as the sole electron acceptor. The isolate was able to grow chemolithoautotrophically with carbon dioxide. Various organic substrates such as complex proteinaceous compounds, carbohydrates, organic acids, amino acids and sugars could also support growth as the carbon source instead of carbon dioxide with sulfur oxidation. The G+C content of the genomic DNA was 43.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate belonged to the genus Thiomicrospira and was most closely related to Thiomicrospira crunogena strain TH-55(T) and Thiomicrospira sp. strain L-12, while DNA-DNA hybridization demonstrated that the novel isolate could be genetically differentiated from previously described strains of Thiomicrospira. On the basis of its physiological and molecular properties the isolate is representative of a novel Thiomicrospira species, for which the name Thiomicrospira thermophila sp. nov. is proposed (type strain, I78(T)=JCM 12397(T)=DSM 16397(T)). PMID- 15545480 TI - Aquiflexum balticum gen. nov., sp. nov., a novel marine bacterium of the Cytophaga-Flavobacterium-Bacteroides group isolated from surface water of the central Baltic Sea. AB - A bacterial isolate from the Baltic Sea, BA160(T), was characterized for its physiological and biochemical features, fatty acid profile, G+C content and phylogenetic position based on 16S rRNA gene sequences. The strain was isolated from the surface water of the central Baltic Sea during the decay of a plankton bloom. Phylogenetic analyses of the 16S rRNA gene sequence revealed a clear affiliation with the family 'Flexibacteraceae', and showed the closest phylogenetic relationship with the species Belliella baltica and Cyclobacterium marinum. The G+C content of the DNA was 38.4 mol%. The strain was red-coloured due to carotenoids, Gram-negative, rod-shaped, and catalase- and oxidase positive. Growth was observed at salinities from 0 to 6 %, with an optimum around 1.5 %. Temperature for growth ranged from 4 to 40 degrees C, with an optimum around 30 degrees C. The fatty acids were dominated by branched-chain fatty acids (>87 %), with a high abundance of iso-C(15 : 0) (23 %) and anteiso-C(15 : 0) (19 %). According to its morphology, physiology, fatty acid composition, G+C content and 16S rRNA gene sequence, strain BA160(T) is considered to represent a new genus of the family 'Flexibacteraceae'. Due to its aquatic origin, the name Aquiflexum balticum gen. nov, sp. nov. is suggested for the type species (type strain, BA160(T)=DSM 16537(T)=LMG 22565(T)=CIP 108445(T)) of the new genus. PMID- 15545481 TI - Emended description of the genus Glycomyces and description of Glycomyces algeriensis sp. nov., Glycomyces arizonensis sp. nov. and Glycomyces lechevalierae sp. nov. AB - A polyphasic taxonomic evaluation of presumptive strains representative of the genus Glycomyces held within the Agricultural Research Service Culture Collection resulted in the discovery of three novel species. Analysis of the whole cell sugar content of these novel species, as well as of two species presently assigned to the genus, revealed that the whole cell sugar pattern was different from that reported in the formal description of the genus Glycomyces. The sugars present in all strains studied included ribose, xylose, mannose and galactose rather than xylose and arabinose as reported in the original description of the genus. Moreover, the menaquinone patterns observed for the novel species also deviated from the original genus description. The formal description of the genus Glycomyces is emended to reflect these new data. The novel species proposed and described are Glycomyces algeriensis sp. nov. (type strain NRRL B-16327(T)=DSM 44727(T)), Glycomyces arizonensis sp. nov. (type strain NRRL B-16153(T)=DSM 44726(T)) and Glycomyces lechevalierae sp. nov. (type strain NRRL B-16149(T)=DSM 44724(T)). PMID- 15545482 TI - Shewanella marisflavi sp. nov. and Shewanella aquimarina sp. nov., slightly halophilic organisms isolated from sea water of the Yellow Sea in Korea. AB - Two Gram-negative, motile, non-spore-forming, rod-shaped organisms, strains SW 117(T) and SW-120(T), were isolated from sea water of the Yellow Sea in Korea and subjected to a polyphasic taxonomic study. Strains SW-117(T) and SW-120(T) simultaneously contained both menaquinones (MK) and ubiquinones (Q) as isoprenoid quinones; the predominant menaquinone was MK-7 and the predominant ubiquinones were Q-7 and Q-8. The major fatty acid detected in the two strains was iso-C(15 : 0). The DNA G+C content of strains SW-117(T) and SW-120(T) was 51 and 54 mol%, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that strains SW-117(T) and SW-120(T) fall within the radiation of the cluster comprising Shewanella species. Strains SW-117(T) and SW-120(T) showed a 16S rRNA gene sequence similarity of 97.4 % and a DNA-DNA relatedness level of 10.1 %. Strains SW-117(T) and SW-120(T) exhibited 16S rRNA gene sequence similarity levels of 93.8-98.5 % and 92.4-97.0 %, respectively, to Shewanella species. Strain SW-117(T) exhibited DNA-DNA relatedness levels of 8.3-20.3 % to the type strains of six phylogenetically related Shewanella species. On the basis of phenotypic, phylogenetic and genetic data, strains SW-117(T) and SW-120(T) were classified in the genus Shewanella as two distinct novel species, for which the names Shewanella marisflavi sp. nov. (type strain, SW-117(T)=KCCM 41822(T)=JCM 12192(T)) and Shewanella aquimarina sp. nov. (type strain, SW-120(T)=KCCM 41821(T)=JCM 12193(T)) are proposed, respectively. PMID- 15545483 TI - Thermosinus carboxydivorans gen. nov., sp. nov., a new anaerobic, thermophilic, carbon-monoxide-oxidizing, hydrogenogenic bacterium from a hot pool of Yellowstone National Park. AB - A new anaerobic, thermophilic, facultatively carboxydotrophic bacterium, strain Nor1(T), was isolated from a hot spring at Norris Basin, Yellowstone National Park. Cells of strain Nor1(T) were curved motile rods with a length of 2.6-3 microm, a width of about 0.5 microm and lateral flagellation. The cell wall structure was of the Gram-negative type. Strain Nor1(T) was thermophilic (temperature range for growth was 40-68 degrees C, with an optimum at 60 degrees C) and neutrophilic (pH range for growth was 6.5-7.6, with an optimum at 6.8 7.0). It grew chemolithotrophically on CO (generation time, 1.15 h), producing equimolar quantities of H(2) and CO(2) according to the equation CO+H(2)O- >CO(2)+H(2). During growth on CO in the presence of ferric citrate or amorphous ferric iron oxide, strain Nor1(T) reduced ferric iron but produced H(2) and CO(2) at a ratio close to 1 : 1, and growth stimulation was slight. Growth on CO in the presence of sodium selenite was accompanied by precipitation of elemental selenium. Elemental sulfur, thiosulfate, sulfate and nitrate did not stimulate growth of strain Nor1(T) on CO and none of these chemicals was reduced. Strain Nor1(T) was able to grow on glucose, sucrose, lactose, arabinose, maltose, fructose, xylose and pyruvate, but not on cellobiose, galactose, peptone, yeast extract, lactate, acetate, formate, ethanol, methanol or sodium citrate. During glucose fermentation, acetate, H(2) and CO(2) were produced. Thiosulfate was found to enhance the growth rate and cell yield of strain Nor1(T) when it was grown on glucose, sucrose or lactose; in this case, acetate, H(2)S and CO(2) were produced. In the presence of thiosulfate or ferric iron, strain Nor1(T) was also able to grow on yeast extract. Lactate, acetate, formate and H(2) were not utilized either in the absence or in the presence of ferric iron, thiosulfate, sulfate, sulfite, elemental sulfur or nitrate. Growth was completely inhibited by penicillin, ampicillin, streptomycin, kanamycin and neomycin. The DNA G+C content of the strain was 51.7+/-1 mol%. Analysis of the 16S rRNA gene sequence revealed that strain Nor1(T) belongs to the Bacillus-Clostridium phylum of the Gram positive bacteria. On the basis of the studied phenotypic and phylogenetic features, we propose that strain Nor1(T) be assigned to a new genus, Thermosinus gen. nov. The type species is Thermosinus carboxydivorans sp. nov. (type strain, Nor1(T)=DSM 14886(T)=VKM B-2281(T)). PMID- 15545484 TI - Classification of isolates from locations in Austria and Yellowstone National Park as Geobacillus tepidamans sp. nov. AB - Two moderately thermophilic, Gram-positive, spore-forming bacteria were isolated from different geographical locations and sources; strain GS5-97(T) from a beet sugar factory in Leopoldsdorf, Lower Austria, and strain YNP10 from a geothermally heated soil, Yellowstone National Park, USA. The sequences of their 16S rRNA genes were found to be 99.8% identical, and DNA-DNA hybridization experiments revealed that strains GS5-97(T) and YNP10 share 89.9 mol% similarity to each other, but only 34.3 and 39.2 mol% similarity, respectively, to Geobacillus caldoxylosilyticus DSM 12041(T), which is their closest related type strain. A polyphasic analysis showed that these two isolates were more similar to each other than to other characterized geobacilli. Their DNA G+C content was 43.2 and 42.4 mol%, respectively, and they were identical with respect to many phenotypic features (e.g. T(opt) 55 degrees C; pH(opt) 7.0). Both strains clearly displayed best growth when cultured aerobically. They differed slightly in their cellular fatty acid profiles and polar lipid pattern, and genotypically they could also be distinguished based on randomly amplified polymorphic DNA fingerprints and internal transcribed spacer analysis. Freeze-etching experiments revealed oblique surface layer (S-layer) lattices in both strains, and biochemical analyses of the purified S-layer proteins indicated the occurrence of glycosylation. Based on the properties of these organisms relative to those currently documented for the genus Geobacillus and for the various sister genera in the Bacillus radiation, a novel species is proposed, Geobacillus tepidamans sp. nov., with GS5-97(T) (=ATCC BAA-942(T)=DSM 16325(T)) as the type strain. Strain YNP10 has been deposited in the American Type Culture Collection as ATCC BAA-943. PMID- 15545485 TI - Campylobacter insulaenigrae sp. nov., isolated from marine mammals. AB - Phenotypic and phylogenetic studies were performed on four Campylobacter-like organisms recovered from three seals and a porpoise. Comparative 16S rRNA gene sequencing studies demonstrated that the organisms represent a hitherto unknown subline within the genus Campylobacter, associated with a subcluster containing Campylobacter jejuni, Campylobacter coli and Campylobacter lari. DNA-DNA hybridization studies confirmed that the bacteria belonged to a single species, for which the name Campylobacter insulaenigrae sp. nov. is proposed. The type strain of Campylobacter insulaenigrae sp. nov. is NCTC 12927(T) (=CCUG 48653(T)). PMID- 15545486 TI - Arthrobacter gangotriensis sp. nov. and Arthrobacter kerguelensis sp. nov. from Antarctica. AB - Two coryneform bacteria were isolated from a penguin rookery soil sample collected in Antarctica, near the Indian station Dakshin Gangotri (strain Lz1y(T)), and from sea water from Kerguelen island, Antarctica (strain KGN15(T)). They have morphological and chemotaxonomic properties (peptidoglycan A4alpha type; major menaquinones MK-8, MK-9 and MK-10; predominant fatty acids anteiso C(15 : 0) and anteiso-C(17 : 0)) that are characteristic of members of the genus Arthrobacter. The isolates shared 97.8 % 16S rRNA gene sequence similarity to each other and were most closely related to Arthrobacter sulfureus (about 98.5 % sequence similarity). DNA-DNA hybridization experiments revealed 50 % relatedness between the isolates, while the levels of DNA-DNA relatedness between strains Lz1y(T) and KGN15 (T) and their phylogenetic relative, A. sulfureus, were respectively 54 and 12 %. Based on the above data and distinct phenotypic differences between the isolates and A. sulfureus, two novel species are proposed, Arthrobacter gangotriensis sp. nov. (type strain Lz1y(T)=DSM 15796(T)=JCM 12166(T)) and Arthrobacter kerguelensis sp. nov. (type strain KGN15(T)=DSM 15797(T)=JCM 12165(T)). PMID- 15545487 TI - Alkalibacterium psychrotolerans sp. nov., a psychrotolerant obligate alkaliphile that reduces an indigo dye. AB - A psychrotolerant, obligately alkaliphilic bacterium, IDR2-2(T), which is able to reduce indigo dye was isolated from a fermented polygonum indigo (Polygonum tinctorium Lour.) produced in Date, Hokkaido, using a traditional Japanese method. The isolate grew at pH 9-12 but not at pH 7-8. It was a Gram-positive, facultatively anaerobic, straight rod-shaped bacterium with peritrichous flagella. The isolate grew in 0-17 % (w/v) NaCl but not at NaCl concentrations higher than 18 % (w/v). Its major cellular fatty acids were C(14 : 0), C(16 : 0), C(16 : 1)9c and C(18 : 1)9c, and its DNA G+C content was 40.6 mol%. dl-lactic acid was the major end-product from d-glucose. No quinones could be detected. The peptidoglycan type was A4beta, Orn-d-Glu. A phylogenetic analysis based on 16S rRNA gene sequence data indicated that strain IDR2-2(T) is a member of the genus Alkalibacterium. DNA-DNA hybridization revealed low relatedness (less than 25 %) between the isolate and two phylogenetically related strains, Alkalibacterium olivapovliticus and Marinilactibacillus psychrotolerans. On the basis of phenotypic characteristics, phylogenetic data and DNA-DNA relatedness data, the isolate merits classification as a novel species, for which the name Alkalibacterium psychrotolerans sp. nov. is proposed. The type strain is IDR2 2(T) (=JCM 12281(T)=NCIMB 13981(T)). PMID- 15545488 TI - Mycobacterium cosmeticum sp. nov., a novel rapidly growing species isolated from a cosmetic infection and from a nail salon. AB - Four isolates of a rapidly growing Mycobacterium species had a mycolic acid pattern similar to that of Mycobacterium smegmatis, as determined by HPLC analyses. Three of the isolates were from footbath drains and a sink at a nail salon located in Atlanta, GA, USA; the fourth was obtained from a granulomatous subdermal lesion of a female patient in Venezuela who was undergoing mesotherapy. By random amplified polymorphic DNA electrophoresis and PFGE of large restriction fragments, the three isolates from the nail salon were shown to be the same strain but different from the strain from the patient in Venezuela. Polymorphisms in regions of the rpoB, hsp65 and 16S rRNA genes that were shown to be useful for species identification matched for the two strains but were different from those of other Mycobacterium species. The 16S rRNA gene sequence placed the strains in a taxonomic group along with Mycobacterium frederiksbergense, Mycobacterium hodleri, Mycobacterium diernhoferi and Mycobacterium neoaurum. The strains produced a pale-yellow pigment when grown in the dark at the optimal temperature of 35 degrees C. Biochemical testing showed that the strains were positive for iron uptake, nitrate reduction and utilization of d-mannitol, d-xylose, iso-myo inositol, l-arabinose, citrate and d-trehalose. The strains were negative for d sorbitol utilization and production of niacin and 3-day arylsulfatase, although arylsulfatase activity was observed after 14 days. The isolates grew on MacConkey agar without crystal violet but not on media containing 5 % (w/v) NaCl or at 45 degrees C. They were susceptible to ciprofloxacin, amikacin, tobramycin, cefoxitin, clarithromycin, doxycycline, sulfamethoxazole and imipenem. The name Mycobacterium cosmeticum sp. nov. is proposed for this novel species; two strains, LTA-388(T) (=ATCC BAA-878(T)=CIP 108170(T)) (the type strain) and 2003 11-06 (=ATCC BAA-879=CIP 108169) have been designated, respectively, for the strains of the patient in Venezuela and from the nail salon in Atlanta, GA, USA. PMID- 15545489 TI - Phylum-specific environmental DNA analysis reveals remarkably high global biodiversity of Cercozoa (Protozoa). AB - This study presents the first 18S rRNA multi-library environmental PCR survey of a single protozoan phylum, Cercozoa Cavalier-Smith 1998, from a range of different habitats. Phylogenetic analysis reveals at least nine novel clades within the phylum, several possibly at the level of order or above. Further experiments are described to ascertain the true ecological and geographical distributions of some clades that might be inferred from the tree to be restricted in either or both ways. These results suggest that the diversity of cercozoan taxa may run into thousands of lineages, making it comparable in diversity to the largest better-characterized protozoan phyla, e.g. Ciliophora (ciliates and suctorians) and Foraminifera. New sequences of cultured Spongomonas, Metromonas and Metopion are also presented. In the light of these additions, and the increased taxon sampling from the environmental libraries, some revisions of cercozoan classification are made: the transfer of Spongomonadea from Reticulofilosa to Monadofilosa; the removal of Metopiida from Sarcomonadea; and the creation of the new order Metromonadida, currently containing the single genus Metromonas. Although Metromonas groups with weak to moderate support with Chlorarachnea, it is here placed in superclass Monadofilosa, to which it is morphologically more similar. PMID- 15545490 TI - Candida leandrae sp. nov., an asexual ascomycetous yeast species isolated from tropical plants. AB - The novel yeast species Candida leandrae is described based on eight isolates from decaying fruits of Leandra reversa Cogn. (Melastomataceae) in an Atlantic rainforest site in Brazil, one from a Convolvulaceae flower in Costa Rica and one from a drosophilid in Hawai'i. The strains differed in their colony morphology, one being butyrous and smooth and the other being filamentous and rugose. Sequences of the D1/D2 domains of the large-subunit rRNA gene from both morphotypes were identical. C. leandrae belongs to the Kodamaea clade and is closely related to Candida restingae. The two species can be separated on the basis of growth at 37 degrees C and the assimilation of melezitose, negative in the novel species. The type culture of C. leandrae is strain UNESP 00-64R(T) (=CBS 9735(T)=NRRL Y-27757(T)). PMID- 15545491 TI - Expansion of the Candida tanzawaensis yeast clade: 16 novel Candida species from basidiocarp-feeding beetles. AB - A major clade of new yeast taxa from the digestive tract of basidiocarp-feeding beetles is recognized based on rRNA gene sequence analyses. Almost 30 % of 650 gut isolates formed a statistically well-supported clade that included Candida tanzawaensis. The yeasts in the clade were isolated from 11 families of beetles, of which Tenebrionidae and Erotylidae were most commonly sampled. Repeated isolation of certain yeasts from the same beetle species at different times and places indicated strong host associations. Sexual reproduction was never observed in the yeasts. Based on comparisons of small- and large-subunit rRNA gene sequences and morphological and physiological traits, the yeasts were placed in Candida ambrosiae and in 16 other undescribed taxa. In this report, the novel species in the genus Candida are described and their relationships with other taxa in the Saccharomycetes are discussed. The novel species and their type strains are as follows: Candida guaymorum (NRRL Y-27568(T)=CBS 9823(T)), Candida bokatorum (NRRL Y-27571(T)=CBS 9824(T)), Candida kunorum (NRRL Y-27580(T)=CBS 9825(T)), Candida terraborum (NRRL Y-27573(T)=CBS 9826(T)), Candida emberorum (NRRL Y-27606(T)=CBS 9827(T)), Candida wounanorum (NRRL Y-27574(T)=CBS 9828(T)), Candida yuchorum (NRRL Y-27569(T)=CBS 9829(T)), Candida chickasaworum (NRRL Y 27566(T)=CBS 9830(T)), Candida choctaworum (NRRL Y-27584(T)=CBS 9831(T)), Candida bolitotheri (NRRL Y-27587(T)=CBS 9832(T)), Candida atakaporum (NRRL Y 27570(T)=CBS 9833(T)), Candida panamericana (NRRL Y-27567(T)=CBS 9834(T)), Candida bribrorum (NRRL Y-27572(T)=CBS 9835(T)), Candida maxii (NRRL Y 27588(T)=CBS 9836(T)), Candida anneliseae (NRRL Y-27563(T)=CBS 9837(T)) and Candida taliae (NRRL Y-27589(T)=CBS 9838(T)). PMID- 15545492 TI - Kazachstania aerobia sp. nov., an ascomycetous yeast species from aerobically deteriorating corn silage. AB - In an investigation of the yeast biota involved in silage deterioration, a considerable number of strains belonging to Saccharomyces and related genera were isolated from aerobically deteriorating corn silage in Tochigi, Japan. Analysis of sequences of the internal transcribed spacer and the large-subunit rRNA gene D1/D2 domain and electrophoretic karyotyping indicated that two of the strains, NS 14(T) and NS 26, represent a novel species with close phylogenetic relationships to Kazachstania servazzii and Kazachstania unispora. It is proposed that the novel species be named Kazachstania aerobia sp. nov., with NS 14(T) (=AS 2.2384(T)=CBS 9918(T)) as the type strain. PMID- 15545493 TI - Proposal to acknowledge Beijerinck as the original author of the species Pantoea agglomerans. Request for an opinion. AB - The name 'Bacillus agglomerans' was first published by Beijerinck in 1888. Ewing and Fife changed the name to Enterobacter agglomerans in 1972 as a new combination, acknowledging that Beijerinck had been the original author and making the new full name Enterobacter agglomerans (Beijerinck 1888) Ewing and Fife 1972. Beijerinck's name was omitted from the Approved Lists of Bacterial Names in 1980, which listed only Ewing and Fife as the authors. The current listings in the List of Bacterial Names with Standing in Nomenclature follow the Approved Lists and omit Beijerinck from the names for both Enterobacter agglomerans and Pantoea agglomerans. It is proposed that Beijerinck be acknowledged as the original author of this species by having his name reinstated in its full name, Pantoea agglomerans (Beijerinck 1888) Gavini et al. 1989, and a Request for an Opinion is put forward to the Judicial Commission. PMID- 15545494 TI - Proposal to list ATCC 43642 as the type strain of Leptospira interrogans in the Approved Lists of Bacterial Names. Request for an opinion. AB - ATCC 23581(T) has been cited as the type strain of Leptospira interrogans since 1980. In 1986, the Centers for Disease Control informed the ATCC that this strain was Leptospira interrogans serovar budapest, not serovar icterohaemorrhagiae as deposited originally. An authentic culture of the Leptospira interrogans serovar icterohaemorrhagiae RGA(T) strain was then submitted and assigned ATCC 43642, which was designated as the type strain of Leptospira interrogans in an article by Yasuda et al. [Yasuda et al. (1987). Int J Syst Bacteriol 37, 407-415]. In this Request for an Opinion to the Judicial Commission, it is proposed that a correction be made in the Approved Lists of Bacterial Names to acknowledge ATCC 43642 as the type strain of Leptospira interrogans. PMID- 15545495 TI - A survey of RNA editing in human brain. AB - We have conducted a survey of RNA editing in human brain by comparing sequences of clones from a human brain cDNA library to the reference human genome sequence and to genomic DNA from the same individual. In the RNA sample from which the library was constructed, approximately 1:2000 nucleotides were edited out of >3 Mb surveyed. All edits were adenosine to inosine (A-->I) and were predominantly in intronic and in intergenic RNAs. No edits were found in translated exons and few in untranslated exons. Most edits were in high-copy-number repeats, usually Alus. Analysis of the genome in the vicinity of edited sequences strongly supports the idea that formation of intramolecular double-stranded RNA with an inverted copy underlies most A-->I editing. The likelihood of editing is increased by the presence of two inverted copies of a sequence within the same intron, proximity of the two sequences to each other (preferably within 2 kb), and by a high density of inverted copies in the vicinity. Editing exhibits sequence preferences and is less likely at an adenosine 3' to a guanosine and more likely at an adenosine 5' to a guanosine. Simulation by BLAST alignment of the double-stranded RNA molecules that underlie known edits indicates that there is a greater likelihood of A-->I editing at A:C mismatches than editing at other mismatches or at A:U matches. However, because A:U matches in double-stranded RNA are more common than all mismatches, overall the likely effect of editing is to increase the number of mismatches in double-stranded RNA. PMID- 15545496 TI - Noncoding regulatory sequences of Ciona exhibit strong correspondence between evolutionary constraint and functional importance. AB - We show that sequence comparisons at different levels of resolution can efficiently guide functional analyses of regulatory regions in the ascidians Ciona savignyi and Ciona intestinalis. Sequence alignments of several tissue specific genes guided discovery of minimal regulatory regions that are active in whole-embryo reporter assays. Using the Troponin I (TnI) locus as a case study, we show that more refined local sequence analyses can then be used to reveal functional substructure within a regulatory region. A high-resolution saturation mutagenesis in conjunction with comparative sequence analyses defined essential sequence elements within the TnI regulatory region. Finally, we found a significant, quantitative relationship between function and sequence divergence of noncoding functional elements. This work demonstrates the power of comparative sequence analysis between the two Ciona species for guiding gene regulatory experiments. PMID- 15545497 TI - Coelacanth genome sequence reveals the evolutionary history of vertebrate genes. AB - The coelacanth is one of the nearest living relatives of tetrapods. However, a teleost species such as zebrafish or Fugu is typically used as the outgroup in current tetrapod comparative sequence analyses. Such studies are complicated by the fact that teleost genomes have undergone a whole-genome duplication event, as well as individual gene-duplication events. Here, we demonstrate the value of coelacanth genome sequence by complete sequencing and analysis of the protocadherin gene cluster of the Indonesian coelacanth, Latimeria menadoensis. We found that coelacanth has 49 protocadherin cluster genes organized in the same three ordered subclusters, alpha, beta, and gamma, as the 54 protocadherin cluster genes in human. In contrast, whole-genome and tandem duplications have generated two zebrafish protocadherin clusters comprised of at least 97 genes. Additionally, zebrafish protocadherins are far more prone to homogenizing gene conversion events than coelacanth protocadherins, suggesting that recombination- and duplication-driven plasticity may be a feature of teleost genomes. Our results indicate that coelacanth provides the ideal outgroup sequence against which tetrapod genomes can be measured. We therefore present L. menadoensis as a candidate for whole-genome sequencing. PMID- 15545498 TI - Extensive and breed-specific linkage disequilibrium in Canis familiaris. AB - The 156 breeds of registered dogs in the United States offer a unique opportunity to map genes important in disease susceptibility, morphology, and behavior. Linkage disequilibrium (LD) is of current interest for its application in whole genome association mapping, since the extent of LD determines the feasibility of such studies. We have measured LD at five genomic intervals, each 5 Mb in length and composed of five clusters of sequence variants spaced 800 kb-1.6 Mb apart. These intervals are located on canine chromosomes 1, 2, 3, 34, and 37, and none is under obvious selective pressure. Approximately 20 unrelated dogs were assayed from each of five breeds: Akita, Bernese Mountain Dog, Golden Retriever, Labrador Retriever, and Pekingese. At each genomic interval, SNPs and indels were discovered and typed by resequencing. Strikingly, LD in canines is much more extensive than in humans: D' falls to 0.5 at 400-700 kb in Golden Retriever and Labrador Retriever, 2.4 Mb in Akita, and 3-3.2 Mb in Bernese Mountain Dog and Pekingese. LD in dog breeds is up to 100x more extensive than in humans, suggesting that a correspondingly smaller number of markers will be required for association mapping studies in dogs compared to humans. We also report low haplotype diversity within regions of high LD, with 80% of chromosomes in a breed carrying two to four haplotypes, as well as a high degree of haplotype sharing among breeds. PMID- 15545499 TI - Intraspecies sequence comparisons for annotating genomes. AB - Analysis of sequence variation among members of a single species offers a potential approach to identify functional DNA elements responsible for biological features unique to that species. Due to its high rate of allelic polymorphism and ease of genetic manipulability, we chose the sea squirt, Ciona intestinalis, to explore intraspecies sequence comparisons for genome annotation. A large number of C. intestinalis specimens were collected from four continents, and a set of genomic intervals were amplified, resequenced, and analyzed to determine the mutation rates at each nucleotide in the sequence. We found that regions with low mutation rates efficiently demarcated functionally constrained sequences: these include a set of noncoding elements, which we showed in C. intestinalis transgenic assays to act as tissue-specific enhancers, as well as the location of coding sequences. This illustrates that comparisons of multiple members of a species can be used for genome annotation, suggesting a path for the annotation of the sequenced genomes of organisms occupying uncharacterized phylogenetic branches of the animal kingdom. It also raises the possibility that the resequencing of a large number of Homo sapiens individuals might be used to annotate the human genome and identify sequences defining traits unique to our species. PMID- 15545500 TI - Preventing breast, ovarian cancer in BRCA carriers: rational of prophylactic surgery and promises of surveillance. PMID- 15545501 TI - Appropriate payment will follow quality care. PMID- 15545502 TI - In vivo optical imaging of pleural space drainage to lymph nodes of prognostic significance. AB - BACKGROUND: Understanding the spatial and temporal drainage patterns of the pleural space could have profound impact on the treatment of lung cancer and mesothelioma. The purpose of this study was to identify the in vivo pattern of drainage from the pleural space to prognostic lymph node stations. METHODS: Fifty six rats underwent pleural space injection of a novel lymph tracer composed of recombinant human serum albumin (HSA) covalently conjugated to the near-infrared (NIR) fluorophore IRDye78 via an amide bond (HSA-78). Nodal uptake was imaged at 10, 20, 30, and 60 minutes and 4, 12, and 24 hours after injection with a custom system that simultaneously acquires color video, NIR fluorescence of HSA-78, and a merged picture of the two. Six pigs underwent the same procedure with imaging at 30 minutes, 1 hour, and 24 hours. RESULTS: In both the rat model and the pig model, HSA-78 drained from the pleural space to superior mediastinal lymph nodes first, followed by other intrathoracic and then extrathoracic lymph nodes over the course of 24 hours. CONCLUSION: NIR fluorescence imaging in two species shows that the superior mediastinal lymph nodes are the first to drain the pleural space. Over the course of 24 hours, the pleural space also communicates with other intrathoracic and then extrathoracic lymph nodes. This study also demonstrates an intraoperative method for identifying nodes communicating with the pleural space, with potential utility in the staging and/or resection of lung cancer and mesothelioma. PMID- 15545503 TI - Sentinel lymph node metastases detected by immunohistochemistry only do not mandate complete axillary lymph node dissection in breast cancer. AB - BACKGROUND: The significance of breast cancer sentinel lymph node (SLN) metastases detected only by immunohistochemistry staining (IHC) remains poorly understood. This study attempted to quantify the risk of non-SLN metastases. METHODS: A prospectively collected database of 750 consecutive SLN biopsy procedures in breast cancer patients was reviewed. Medical records were reviewed to supplement the database. RESULTS: SLNs were identified in 738 (98.4%) of these procedures in 723 patients. Of these, 151 patients (20.5%) had metastases detected by hematoxylin and eosin staining (H&E), and 33 (4.6%) of the 718 with known IHC staining results had metastases detected by IHC only. Twenty-eight (84.8%) of 33 patients with IHC-detected metastases underwent complete axillary lymph node dissection (CALND). The median primary tumor size was 2.0 cm among those undergoing CALND and 0.9 cm among the five patients treated without CALND (P = .10). Two of the 28 patients (7.1%) had additional metastases detected with CALND. These patients had a T3 or T4 invasive lobular primary tumor. Of 24 patients with T1 or T2 primary tumors and IHC-detected metastases who underwent CALND, none had additional metastases detected. Median follow-up was 14.5 months. All patients with IHC-detected SLN metastases were treated with adjuvant systemic therapy. None of the five patients with IHC-detected metastases not undergoing CALND has subsequently manifested clinical axillary disease. CONCLUSIONS: CALND could have been or was safely omitted in 29 of 29 patients with T1 or T2 primary tumors and metastases detected by IHC. Such patients should be counseled about this low risk before CALND is recommended. PMID- 15545504 TI - Assessment of the current Medicare reimbursement system for breast cancer operations. AB - BACKGROUND: Medicare determines procedural reimbursement by means of formulas considering physician work, practice, and liability expenses. Since no mechanism exists to consider outcomes in calculating reimbursements, we hypothesized that Medicare reimbursements do not correlate with outcomes for different breast cancer operations. METHODS: We prospectively studied 240 patients with T1, 2N0M0 breast cancer in three surgical treatment arms: segmental mastectomy with axillary node dissection (SM&ALND ; n = 42); SM with sentinel node dissection (SM&SLND ; n = 96); and mastectomy without reconstruction (MRM; n = 102). Outcome parameters of complications, hospital stay, analgesic usage, and days to return to work were correlated with procedure reimbursements. RESULTS: Median follow-up was 26 months. SM&SLND patients rarely required hospital stays (14%) in comparison with either SM&ALND (96%) or MRM patients (99%) (P < 0.001). SM&ALND and MRM patients required 9 and 10 median days of narcotics, respectively, versus 1 day in the SLND group (P < 0.001). SM&SLND patients returned to work at a median of 3 days, in comparison with 19 for SM&ALND and 26 for MRM patients (P < 0.001). Complications were more common in the MRM group (67% numbness/10% pain) and the SM&ALND group (56%/9%) than in the SM&SLND group (0%/1%). Reimbursements were inversely correlated with outcomes. MRM was reimbursed the highest, at an average of 1,075.03 dollars, with SM&ALND at 882.72 dollars. SM&SLND was reimbursed at 642.00 dollars. CONCLUSIONS: Medicare reimbursements for breast cancer operations do not correlate with outcomes. Less-invasive procedures are paid for at lower rates despite better outcomes and fewer complications. The data from this study raise the question of the impact of reimbursement on breast procedure selection. PMID- 15545505 TI - Prediction of lymph node metastasis with use of artificial neural networks based on gene expression profiles in esophageal squamous cell carcinoma. AB - BACKGROUND: The aim of the study was (1) to detect candidate genes involved in lymph node metastasis in esophageal cancers and (2) to investigate whether we can estimate and predict occurrence of lymph node metastasis by analyzing artificial neural networks (ANNs) using these gene subsets. METHODS: Twenty-eight primary esophageal squamous cell carcinomas were used. Gene expression profiles of all primary tumors were obtained by cDNA microarray. Lymph node metastasis-related genes were extracted with use of Significance Analysis of Microarrays (SAM). Predictive accuracy for lymph node metastasis was calculated by evaluation of 28 cases by ANNs with leave-one-out cross-n. The results were compared with those of other analyses such as clustering or predictive scoring (LMS). RESULTS: Our ANN model could predict lymph node metastasis most accurately with 60 clones. The highest predictive accuracy for lymph node metastasis by ANN was 10 of 13 (77%) in newly added cases that were not used for gene selection by SAM and 24 of 28 (86%) in all cases (sensitivity: 15/17, 88%; specificity: 9/11, 82%). Predictive accuracy of LMS was 9 of 13 (69%) in newly added cases and 24 of 28 (86%) in all cases (sensitivity: 17/17, 100%; specificity: 7/11, 67%). It was difficult to extract useful information for the prediction of lymph node metastasis by clustering analysis. CONCLUSIONS: ANN had superior potential in comparison with other methods of analysis for the prediction of lymph node metastasis. This systematic analysis combining SAM with ANN was very useful for the prediction of lymph node metastasis in esophageal cancers and could be applied clinically in the near future. PMID- 15545506 TI - Extracorporeal high intensity focused ultrasound ablation in the treatment of patients with large hepatocellular carcinoma. AB - BACKGROUND: High intensity focused ultrasound (HIFU) is a noninvasive treatment modality that induces complete coagulative necrosis of a deep tumor through the intact skin. The current study was conducted to determine the safety, efficacy, and feasibility of extracorporeal HIFU in the treatment of patients with hepatocellular carcinoma (HCC). METHODS: A total of 55 patients with HCC with cirrhosis were enrolled in this prospective, nonrandomized clinical trial. Among them, 51 patients had unresectable HCC. Tumor size ranged from 4 to 14 cm in diameter with mean diameter of 8.14 cm. According to tumor, node, metastasis (TNM) classification, 15 patients corresponded to stage II, 16 to stage IIIA, and 24 to IIIC. All patients had HIFU, and the median number of HIFU session was 1.69. Safety and efficacy of HIFU were assessed in this trial. RESULTS: No severe side effect was observed in the patients treated with HIFU. Follow-up imaging showed an absence of tumor vascular supply and the shrinkage of treated lesions. Serum alpha-fetoprotein returned to normal level in 34% of patients. The overall survival rates at 6, 12, and 18 months were 86.1%, 61.5%, and 35.3%, respectively. The survival rates were significantly higher in patients in stage II than those in stage IIIA (P = .0132) and in stage IIIC (P = .0265). CONCLUSION: As a noninvasive therapy, HIFU appears to be effective, safe, and feasible in the treatment of patients with HCC. It may play an important role in the ablation of large tumors. PMID- 15545507 TI - Transcriptional regulation of renin: an update. AB - Renin, as a component of the renin-angiotensin system, plays important roles in the regulation of blood pressure, electrolyte homeostasis, and mammalian renal development. Transcription of renin genes is subject to complex developmental and tissue-specific regulation. Progress has been made recently in elucidating the molecular mechanisms involved in renin gene expression. Using mouse As4.1 cells, which have many features characteristic of the renin-expressing juxtaglomerular cells of kidney, a proximal promoter region (-197 to -50 bp) and an enhancer ( 2866 to -2625 bp) have been identified in the mouse renin gene, Ren-1(c), that are critical for its expression. The proximal promoter region contains at least 7 transcription factor-binding sites, including a binding site for the products of Hox, developmental control genes. The enhancer consists of at least 11 transcription factor-binding sites and is responsive to various signal transduction pathways, including cAMP, retinoic acid, endothelin-1, and cytokines, to alter renin mRNA levels. Sequence highly homologous to the mouse enhancer is also found in the human and rat renin genes. How these regulatory regions function in vivo will be the focus of future study. PMID- 15545508 TI - Treating osteoarthritis with cyclooxygenase-2-specific inhibitors: what are the benefits of avoiding blood pressure destabilization? AB - Osteoarthritis and hypertension are highly prevalent among older Americans. Anti inflammatory medications can destabilize blood pressure control. We estimated the decreased cardiovascular risk, premature mortality, and direct health care costs that could be avoided if blood pressure control is not destabilized among hypertensive Americans taking cyclooxygenase-2 (COX-2)-specific inhibitors for osteoarthritis. Data from the Third National Health and Nutrition Examination Survey (NHANES III) provided the distribution of cardiovascular risk factors among American adults with osteoarthritis and hypertension. The Cardiovascular Disease Life Expectancy Model was used to estimate the impact of a 2.26% increase in systolic blood pressure on the basis of results of a randomized trial comparing COX-2-specific inhibitors. A similar analysis was completed for American adults with osteoarthritis and untreated hypertension (> or =140/90 mm Hg). Among 7.3 million Americans with treated hypertension, maintaining blood pressure control would avoid >30,000 stroke deaths and 2,000 coronary deaths resulting in >449,000 person years of life saved and 1.4 billion dollars in direct health care cost savings. When an additional 3.8 million Americans with untreated hypertension are considered, maintaining blood pressure control could prevent >47,000 stroke deaths, 39,000 coronary deaths, and result in 668,000 person years of life saved and >2.4 billion dollars in direct health care cost savings. We conclude that even a small increase in systolic blood pressure among hypertensive Americans with osteoarthritis may substantially increase the clinical and economic burden of cardiovascular disease. Maintaining blood pressure control may be associated with substantial benefits. PMID- 15545509 TI - Pulse pressure and coronary atherosclerosis progression in postmenopausal women. AB - Pulse pressure, an index of large artery stiffness, has been associated with coronary events. However, mechanisms for this association remain unclear. In this study, we examined the relationship between pulse pressure and the progression of coronary atherosclerosis and the effects of hormone replacement therapy (HRT) on pulse pressure in postmenopausal women with angiographically confirmed coronary disease followed for 3.2 years in the Estrogen Replacement in Atherosclerosis (ERA) trial. In the ERA trial, 309 postmenopausal women (mean age 66+/-7 years) with coronary disease were randomized to estrogen, estrogen plus progestin, or placebo, and followed for 3.2 years. Ten standardized epicardial segments were measured for minimal diameter values at baseline and follow-up using quantitative coronary angiography. For this study, mixed-model analysis of covariance was used to: (1) test the association between pulse pressure and change in mean minimum diameter (MMD) adjusted for baseline MMD and (2) the effect of HRT on follow-up pulse pressure. After adjustment for potential confounders, there was a significant graded increase in progression of coronary stenosis with increasing quartiles of baseline pulse pressure (P test for trend=0.0001). The progression rate in women with the highest quartile of baseline pulse pressure was 5-fold higher than in women in the lowest quartile (P<0.01). In postmenopausal women with coronary disease, increased levels of baseline pulse pressure are associated with subsequent progression of coronary atherosclerosis in postmenopausal women. HRT had no detectable effect on pulse pressure. PMID- 15545510 TI - Angiotensin-(1-7) attenuates neointimal formation after stent implantation in the rat. AB - Angiotensin-(1-7) is an endogenous, biologically active peptide of the renin angiotensin system with vasodilatory, antithrombotic, and antiproliferative properties. This study examined the effects of angiotensin-(1-7) infusion on neointimal formation after stent placement in the rat. Male Wistar rats underwent stent implantation in the abdominal aorta or sham surgery. Subsequently, an osmotic minipump was placed for angiotensin-(1-7) (24 microg/kg per hour) or saline administration. After 4 weeks, histomorphometric and histological analyses were performed, and the endothelial function was measured in isolated thoracic aortic rings. Stent implantation resulted in equal mean injury scores within the groups. The angiotensin-(1-7)-treated group displayed a significant reduction in neointimal thickness (112+/-8 versus 141+/-11 microm; P<0.05), neointimal area (0.51+/-0.05 versus 0.70+/-0.07 mm2; P<0.05), and percentage stenosis (10.4+/-1.0 versus 14.0+/-1.3%; P<0.05) compared with the saline-treated group. Furthermore, angiotensin-(1-7) infusion attenuated the stenting-induced impairment in endothelium-dependent relaxation (42.6+/-3.0 versus 64.5+/-6.0% of phenylephrine maximal contraction; P<0.001). In conclusion, angiotensin-(1-7) treatment attenuates neointimal formation after stent implantation in the rat, combined with an improvement of endothelial function. PMID- 15545511 TI - Role of podocytes for reversal of glomerulosclerosis and proteinuria in the aging kidney after endothelin inhibition. AB - The cause of focal-segmental glomerulosclerosis as a consequence of physiological aging, which is believed to be inexorable, is unknown. This study investigated whether inhibition of endothelin-1, a growth-promoting peptide contributing to renal injury in hypertension and diabetes, affects established glomerulosclerosis and proteinuria in the aged kidney. We also determined the role of endothelin receptors for podocyte injury in vivo and in vitro. Aged Wistar rats, a model of spontaneous age-dependent glomerulosclerosis, were treated with the orally active endothelin subtype A (ET(A)) receptor antagonist darusentan, and evaluation of renal histology, renal function studies, and expression analyses were performed. In vitro experiments using puromycin aminonucleoside to induce podocyte injury investigated the role of ET(A) receptor signaling for apoptosis, cytoskeletal injury, and DNA synthesis. In aged Wistar rats, established glomerulosclerosis and proteinuria were reduced by >50% after 4 weeks of darusentan treatment, whereas blood pressure, glomerular filtration rate, or tubulo-interstitial renal injury remained unaffected. Improvement of structural injury in glomeruli and podocytes was accompanied by a reduction of the expression of matrix metalloproteinase-9 and p21Cip1/WAF1. In vitro experiments blocking ET(A) receptors using specific antagonists or RNA interference prevented apoptosis and structural damage to podocytes induced by puromycin aminonucleoside. In conclusion, these results support the hypothesis that endogenous endothelin contributes to glomerulosclerosis and proteinuria in the aging kidney. The results further suggest that age-dependent glomerulosclerosis is not merely a "degenerative" but a reversible process locally confined to the glomerulus involving recovery of podocytes from previous injury. PMID- 15545512 TI - Novel approach to examining first cardiovascular events after hypertension onset. AB - Hypertension confers risk for multiple types of cardiovascular events, but competing risks for these outcomes are unknown. We estimated the competing risks over 12 years after hypertension onset among cases and age-, sex-, and examination-matched controls using competing Cox cumulative incidence and proportional hazards models. We included all Framingham Heart Study subjects examined after 1977 with new-onset hypertension who were free of cardiovascular disease. There were 645 men and 702 women with new-onset hypertension (mean age: men, 55+/-12 years; women, 59+/-12 years). Compared with matched nonhypertensive controls, subjects with new-onset hypertension were more likely to experience a cardiovascular event first rather than noncardiovascular death. Among new-onset hypertensives, the 12-year competing cumulative incidence of any cardiovascular end point as a first event in men was 24.7%, compared with 9.8% for noncardiovascular death (hazards ratio [HR], 2.53; 95% confidence interval [CI], 1.83 to 3.50); in women, the competing incidences were 16.0% versus 10.1%, respectively (HR, 1.58; 95% CI, 1.13 to 2.20). The most common first major cardiovascular events were hard coronary disease (8.2%) in men and stroke (5.2%) in women. Type and incidence of first cardiovascular events varied by age and severity of hypertension at onset, with stroke predominating among older subjects with new-onset hypertension. After hypertension onset, cardiovascular events are more likely to occur first as opposed to noncardiovascular death. Types of initial events differ by gender, age, and severity of hypertension at onset. These results represent a novel approach to understanding the complications of hypertension and may help target therapies for patients with new-onset hypertension to optimize prevention strategies. PMID- 15545513 TI - Cardiovascular, renal, and metabolic responses to chronic central administration of agouti-related peptide. AB - Although excess hypothalamic agouti-related peptide (AGRP), an endogenous antagonist of the melanocortin 3/4 receptor, causes hyperphagia and obesity, its role in regulating cardiovascular function is unclear. This study examined control of mean arterial pressure (MAP), heart rate (HR), and metabolism during chronic central administration of AGRP in rats. A cannula was placed in the lateral ventricle for intracerebroventricular infusion, and arterial and venous catheters were implanted for monitoring MAP and HR 24 hours per day, as well as intravenous infusions. After a 5-day control period, rats received AGRP (n=6; 0.02 nmol per hour ICV) or artificial cerebrospinal fluid (aCSF; n=9; 0.02 nmol per hour ICV) for 12 days, followed by a 5-day recovery period. A third group was infused intracerebroventricularly with AGRP and pair-fed to match food intake of control rats (n=7). AGRP produced a peak decrease in MAP and HR of -7+/-2 mm Hg and -68+/-7 bpm, respectively, despite increased food intake (from 23+/-0.5 to 36+/-3 g per day) and weight gain (from 350+/-8 to 454+/-5 g). AGRP also increased glomerular filtration rate, plasma insulin, glucose, and leptin. AGRP infusion in pair-fed rats produced a peak decrease in HR of -70+/-8 bpm but did not alter MAP or other variables. The metabolic effects of AGRP may be secondary to hyperphagia because they were abolished in pair-fed rats. aCSF infusion did not change any of the variables studied. These results demonstrate that increased central nervous system AGRP levels produce chronic reductions in MAP and HR despite marked increases in food intake and weight gain that normally tend to raise arterial pressure. PMID- 15545514 TI - Inhibition of sphingomyelin synthesis reduces atherogenesis in apolipoprotein E knockout mice. AB - BACKGROUND: In clinical studies, sphingomyelin (SM) plasma levels correlated with the occurrence of coronary heart disease independently of plasma cholesterol levels. We hypothesized that inhibition of SM synthesis would have antiatherogenic effects. To test this hypothesis, apolipoprotein E (apoE) knockout (KO) mice were treated with myriocin, a potent inhibitor of serine palmitoyltransferase, the rate-limiting enzyme in SM biosynthesis. METHODS AND RESULTS: Diet-admix treatment of apoE-KO mice with myriocin in Western diet for 12 weeks lowered SM and sphinganine plasma levels. Decreases in sphinganine and SM concentrations were also observed in the liver and aorta of myriocin-treated animals compared with controls. Inhibition of de novo sphingolipid biosynthesis reduced total cholesterol and triglyceride plasma levels. Cholesterol distribution in lipoproteins demonstrated a decrease in beta-VLDL and LDL cholesterol and an increase in HDL cholesterol. Oil red O staining of total aortas demonstrated reduction of atherosclerotic lesion coverage in the myriocin treated group. Atherosclerotic plaque area was also reduced in the aortic root and brachiocephalic artery. CONCLUSIONS: Inhibition of de novo SM biosynthesis in apoE-KO mice lowers plasma cholesterol and triglyceride levels, raises HDL cholesterol, and prevents development of atherosclerotic lesions. PMID- 15545515 TI - Elastin degradation and calcification in an abdominal aorta injury model: role of matrix metalloproteinases. AB - BACKGROUND: Elastin calcification is a widespread feature of vascular pathology, and circumstantial evidence exists for a correlation between elastin degradation and calcification. We hypothesized that matrix metalloproteinase (MMP)-mediated vascular remodeling plays a significant role in elastin calcification. METHODS AND RESULTS: In the present studies, we determined that short-term periadventitial treatment of the rat abdominal aorta with low concentrations of calcium chloride (CaCl2) induced chronic degeneration and calcification of vascular elastic fibers in the absence of aneurysm formation and inflammatory reactions. Furthermore, the rate of progression of calcification depended on the application method and concentration of CaCl2 applied periarterially. Initial calcium deposits, associated mainly with elastic fibers, were persistently accompanied by elastin degradation, disorganization of aortic extracellular matrix, and moderate levels of vascular cell apoptosis. Application of aluminum ions (known inhibitors of elastin degradation) before the CaCl2-mediated injury significantly reduced elastin calcification and abolished both extracellular matrix degradation and apoptosis. We also found that MMP-knockout mice were resistant to CaCl2-mediated aortic injury and did not develop elastin degeneration and calcification. CONCLUSIONS: Collectively, these data strongly indicate a correlation between MMP-mediated elastin degradation and vascular calcification. PMID- 15545516 TI - Overexpression of brain natriuretic peptide facilitates neutrophil infiltration and cardiac matrix metalloproteinase-9 expression after acute myocardial infarction. AB - BACKGROUND: Recent clinical trials have shown that systemic infusion of nesiritide, a recombinant human brain natriuretic peptide (BNP), improves hemodynamic parameters in acutely decompensated hearts. This suggests that BNP exerts a direct cardioprotective effect and might thus be a useful therapeutic agent with which to treat acute myocardial infarction (MI). In the present study, we used BNP-transgenic (BNP-Tg) mice with elevated plasma BNP to determine whether and how BNP contributes to left ventricular remodeling and healing after MI. METHODS AND RESULTS: We examined the accumulation of neutrophils and the expression and activation of matrix metalloproteinase (MMP)-9 in the ventricles of male BNP-Tg mice and their nontransgenic (non-Tg) littermates during the early phase after acute MI. The numbers of neutrophils infiltrating the infarcted area were significantly increased in BNP-Tg mice 3 days after MI. In addition, both the gene expression and zymographic activity of MMP-9, but not MMP-2, were significantly higher in BNP-Tg than non-Tg mice. Double immunostaining revealed that neutrophils are the main source of the MMP-9, although doxycycline, an MMP inhibitor, had no effect on neutrophil infiltration of the infarcted area in BNP Tg mice. CONCLUSIONS: These results demonstrate that elevated plasma BNP facilitates neutrophil infiltration of the infarcted area after MI and increases the activity of the MMP-9 they produce. This suggests that BNP plays a key role in the processes of extracellular matrix remodeling and wound-healing during the early phase after acute MI. PMID- 15545517 TI - Dynamin-2 regulates oxidized low-density lipoprotein-induced apoptosis of vascular smooth muscle cell. AB - BACKGROUND: On exposure to oxidized low-density lipoprotein (oxLDL), vascular cells generally undergo apoptosis, which is one of the major pathogenic factors of atherosclerosis. In this study, we examined the role of dynamin (a crucial GTPase protein in endocytosis) in oxLDL-induced apoptosis of vascular smooth muscle cells (VSMC). METHODS AND RESULTS: After oxLDL stimulation, dynamin-2 colocalized with LOX-1 around the cell surface, as well as oxLDL in the cytoplasm, suggesting that dynamin-2 was involved in scavenger receptor-mediated oxLDL endocytosis. Downregulation of dynamin-2 induced by dynamin-2 dominant negative plasmid (K44A) resulted in a decrease of oxLDL uptake and thereby in a reduction of apoptosis. These data demonstrated that dynamin-2 was involved in oxLDL-induced apoptosis via the oxLDL endocytotic pathway. On the other hand, dynamin-2 wild-type plasmid transfection promoted oxLDL-induced apoptosis without increasing oxLDL uptake. Interestingly, the p53 inhibitor pifithrin-alpha (PFT) significantly reduced apoptosis promoted by wild-type dynamin-2 (78% reduction compared with the PFT[-] condition). These results indicated that dynamin-2 enhanced oxLDL-induced apoptosis of VSMC by participating in the p53 pathway, probably as a signal transducer. Moreover, we demonstrated that, in advanced plaques of apolipoprotein E-/- mice, dynamin-2 expression was often enhanced in apoptotic VSMC, suggesting that dynamin-2 might participate in apoptosis of VSMC even in vivo. CONCLUSIONS: Our data demonstrated that dynamin-2 at least partially regulated oxLDL-induced apoptosis of VSMC by participating in 2 independent pathways: the oxLDL endocytotic pathway and the p53 pathway. These findings suggest that dynamin-2 may serve as a new research or therapeutic target in vascular disease. PMID- 15545518 TI - Temporal and spatial variations in structural protein expression during the progression from stunned to hibernating myocardium. AB - BACKGROUND: Dysfunctional and normally perfused remote regions show equal myolysis and glycogen accumulation in pig hibernating myocardium. We tested the hypothesis that these arose secondary to elevations in preload rather than ischemia. METHODS AND RESULTS: Expression of structural protein (desmin, desmoplakin, titin, cardiotin, alpha-smooth muscle actin, lamin-A/C, and lamin B2) in viable dysfunctional myocardium was analyzed by immunohistochemistry. We performed blinded analysis of paired dysfunctional left anterior descending coronary artery and normal remote subendocardial samples from stunned (24 hours; n=6), and hibernating (2 weeks; n=6) myocardium versus sham controls pigs (n=7). Within 24 hours, cardiac myocytes globally reexpressed alpha-smooth muscle actin. In stunned myocardium, cardiotin was globally reduced, whereas reductions in desmin were restricted to the dysfunctional region. Alterations progressed with the transition to hibernating myocardium, in which desmin, cardiotin, and titin were globally reduced. A qualitatively similar reorganization of cytoskeletal proteins occurred 3 hours after transient elevation of left ventricular end diastolic pressure to 33+/-3 mm Hg. CONCLUSIONS: Qualitative cardiomyocyte remodeling similar to that in humans with chronic hibernation occurs rapidly after a critical coronary stenosis is applied, as well as after transient elevations in left ventricular end-diastolic pressure in the absence of ischemia. Thus, reorganization of cytoskeletal proteins in patients with viable dysfunctional myocardium appears to reflect chronic and/or cyclical elevations in preload associated with episodes of spontaneous regional ischemia. PMID- 15545519 TI - Resistin promotes smooth muscle cell proliferation through activation of extracellular signal-regulated kinase 1/2 and phosphatidylinositol 3-kinase pathways. AB - BACKGROUND: Resistin, a novel adipokine, is elevated in patients with type 2 diabetes and may play a role in the vascular complications of this disorder. One recent study has shown that resistin has a proinflammatory effect on endothelial cells. However, there is no information on whether resistin could also affect vascular smooth muscle cells (SMCs). Thus, the purpose of this study was to assess whether resistin could induce SMC proliferation and to study the mechanisms whereby resistin signals in SMCs. METHODS AND RESULTS: Human aortic smooth muscle cells (HASMCs) were stimulated with increasing concentrations of resistin for 48 hours. Cell proliferation was induced by resistin in a dose dependent manner as assessed by direct cell counting. To gain more insights into the mechanism of action of resistin, we investigated the extracellular signal regulated kinase (ERK) and/or phosphatidylinositol 3-kinase (PI3K) signaling pathways. Transient phosphorylation of the p42/44 mitogen-activated protein kinase (ERK 1/2) occurred after addition of resistin to HASMCs. U0126, a specific inhibitor of ERK phosphorylation, significantly inhibited ERK 1/2 phosphorylation and reduced resistin-simulated proliferation of HASMCs. LY294002, a specific PI3K inhibitor, also significantly inhibited HASMC proliferation after resistin stimulation. CONCLUSIONS: Our results demonstrate that resistin induces HASMC proliferation through both ERK 1/2 and Akt signaling pathways. The proliferative action exerted by resistin on HASMCs may account in part for the increased incidence of restenosis in diabetes patients. PMID- 15545520 TI - Enhanced arteriogenesis and wound repair in dystrophin-deficient mdx mice. AB - BACKGROUND: The absence of functional dystrophin in Duchenne muscular dystrophy (DMD) patients and in mdx mice results in progressive muscle degeneration associated with necrosis, fibrosis, and inflammation. Because vascular supply plays a key role in tissue repair, we examined whether new blood vessel development was altered in mdx mice. METHODS AND RESULTS: In a model of hindlimb ischemia on femoral artery dissection, hindlimb perfusion, measured by laser Doppler imaging, was higher in mdx mice (0.67+/-0.26) than in wild-type (WT) mice (0.33+/-0.18, P<0.03). In keeping with these data, a significant increase in arteriole length density was found in mdx mice (13.6+/-8.4 mm/mm3) compared with WT mice (7.8+/-4.6 mm/mm3, P<0.03). Conversely, no difference was observed in capillary density between mice of the 2 genotypes. The enhanced regenerative response was not limited to ischemic skeletal muscle, because in a wound-healing assay, mdx mice showed an accelerated wound closure rate compared with WT mice. Moreover, a vascularization assay in Matrigel plugs containing basic fibroblast growth factor injected subcutaneously revealed an increased length density of arterioles in mdx (46.9+/-14.7 mm/mm3) versus WT mice (19.5+/-5.8 mm/mm3, P<0.001). Finally, serum derived from mdx mice sustained formation of endothelium derived tubular structures in vitro more efficiently than WT serum. CONCLUSIONS: These results demonstrate that arteriogenesis is enhanced in mdx mice both after ischemia and skin wounding and in response to growth factors. PMID- 15545521 TI - High-density lipoprotein stimulates myocardial perfusion in vivo. AB - BACKGROUND: Several clinical studies have demonstrated a close association between plasma HDL cholesterol levels and endothelium-dependent vasodilation in peripheral arteries. In isolated arteries, HDL has been shown to mediate vasodilation via NO release. In vivo, administration of reconstituted HDL restored abnormal endothelial function of the brachial artery in hypercholesterolemic patients. However, no data are currently available on the effect of HDL on myocardial perfusion. METHODS AND RESULTS: In this study, administration of human HDL enhanced incorporation of the perfusion tracer 99mTc methoxyisobutylisonitrile (99mTc-MIBI) into the murine heart in vivo by approximately 18%. This increase was completely abolished in mice deficient for endothelial NO synthase. Because we have recently identified sphingosine 1 phosphate (S1P) as an important vasoactive component contained in HDL, we measured myocardial perfusion after administration of S1P in vivo. We observed an approximately 25% decrease in myocardial MIBI uptake, which was abolished in mice deficient for the S1P receptor S1P3. In S1P3-/- mice, the stimulatory effect of HDL on myocardial perfusion was preserved. CONCLUSIONS: HDL increased myocardial perfusion under basal conditions in vivo via NO-dependent mechanisms, whereas S1P inhibited myocardial perfusion through the S1P3 receptor. Thus, HDL may reduce coronary risk via direct NO-mediated vasodilatory effects on the coronary circulation. PMID- 15545522 TI - Directed vascular expression of human cysteinyl leukotriene 2 receptor modulates endothelial permeability and systemic blood pressure. AB - BACKGROUND: The proinflammatory and vascular actions of cysteinyl leukotrienes (CysLTs) are mediated by 2 receptors: cysteinyl leukotriene 1 receptor (CysLT1R) and cysteinyl leukotriene 2 receptor (CysLT2R). However, the distinct contribution of CysLT2R to the vascular actions of CysLTs has not been addressed. METHODS AND RESULTS: We generated an endothelial cell-specific human CysLT2R (EC hCysLT2R) transgenic (TG) mouse model using the Tie2 promoter/enhancer. Strong expression of hCysLT2R in TG lung and endothelial cells, detected by real-time polymerase chain reaction, markedly enhanced CysLT-stimulated intracellular calcium mobilization compared with endogenous expression in cells from nontransgenic mice. The permeability response to exogenous LTC4 and to endogenous CysLTs evoked by passive cutaneous anaphylaxis was augmented in TG mice. The rapid, systemic pressor response to intravenous LTC4 was also diminished in TG mice coincidentally with augmented production of nitric oxide. CONCLUSIONS: The development of EC-hCysLT2R mice has permitted detection of distinct vascular effects of CysLTs, which can be mediated via the CysLT2R in vivo. PMID- 15545523 TI - Images in cardiovascular medicine. Isolated right ventricular infarction resulting from occlusion of a nondominant right coronary artery. PMID- 15545524 TI - Images in cardiovascular medicine. Double-chambered left ventricle: complete characterization by cardiac magnetic resonance and multidetector-row computed tomography. PMID- 15545525 TI - Letter regarding article by Lazar et al, "Tight glycemic control in diabetic coronary artery bypass graft patients improves perioperative outcomes and decreases recurrent ischemic events". PMID- 15545526 TI - Circulating progenitor cells: search for an identity. PMID- 15545527 TI - Which inhibitor of the renin-angiotensin system should be used in chronic heart failure and acute myocardial infarction? PMID- 15545528 TI - Rejuvenating the aging face. PMID- 15545529 TI - Radiofrequency treatment for middle and lower face laxity. AB - OBJECTIVE: To compare the effectiveness of 1 and 2 radiofrequency (RF) treatments with the ThermaCool TC system (Thermage Inc, Hayward, Calif) on middle and lower face laxity. METHODS: Twenty patients with mild to moderate laxity of the middle and lower face were randomly assigned to receive either a single RF treatment or 2 treatments spaced 1 month apart. Treatment energy levels were titrated to patient tolerance and ranged from 85 to 135 J/cm(2). Acute clinical response was recorded after each session. Standardized photographs were taken before treatment and at 1 and 4 months after the last treatment. Using a percentage scale, 4 blinded physicians experienced in dermatologic laser therapy independently rated improvement in nasolabial folds, marionette lines, jowls, laxity under the chin, and overall appearance. In addition, subjects completed quality-of-life surveys 1 and 4 months after treatment. Each patient paid the same fee for involvement in the study. RESULTS: Eleven patients received a single RF treatment, and 9 patients underwent 2 treatments. All subjects experienced mild edema and mild to moderate erythema as an acute clinical response; no patients experienced burns, skin breakdown, or scarring. At 4-month follow-up, patients in the 2-treatment group received higher scores in all categories of photographic analysis; the difference in improvement in the nasolabial folds was statistically significant (P = .04). In self-assessment ratings, individuals receiving 2 treatments reported more improvement than subjects in the single-treatment group 4 months after treatment (P = .03). In both treatment groups, physician photographic assessment demonstrated continued improvement in all subsites between the 1-month and 4-month assessments (P<.05). Although the overall change noted by both patients and physicians was modest in most patients, 75% of subjects (n = 15) stated they would consider paying for additional treatments. CONCLUSIONS: Two RF treatments yielded significantly better improvement than a single treatment in the nasolabial folds. Significant improvement in laxity after treatment was seen between the 1- and 4-month follow-up visits in both single- and 2-treatment groups. Although overall improvements were modest in both groups, patient satisfaction was relatively high. PMID- 15545530 TI - Dose response of combination photorejuvenation using intense pulsed light activated photodynamic therapy and radiofrequency energy. AB - OBJECTIVE: To report the results of a dose-response study using a novel photorejuvenation regimen consisting of intense pulsed light-activated photodynamic therapy and radiofrequency energy in patients with Fitzpatrick skin types I through III. METHODS: A combination intense pulsed light and radiofrequency device (Syneron Medical Ltd, Yokneam, Israel) was used in combination with the topical photosensitizer aminolevulinic acid (ALA) (Levulan Kerastick; DUSA Pharmaceuticals Inc, Wilmington, Mass). Duplicate titration trials were performed on the dorsal forearm skin of 3 patients with Fitzpatrick skin types I through III. Multiple treatments, with varying times of application of ALA (30 minutes and 1, 2, and 3 hours) and intense pulsed light fluence (24-30 J/cm(2)), were completed to determine a minimal erythema dose and a maximal tolerated dose based on epidermal reaction (erythema and crusting) and patient discomfort. Radiofrequency energy levels were constant in all treatment groups. RESULTS: The ALA application time of 1 to 2 hours for all skin types and fluence levels of 26 to 28 and 24 to 26 J/cm(2) for Fitzpatrick skin types I/II and III, respectively, were determined to be the minimal erythema dose. The ALA application times of 2 to 3 hours and fluence levels of 28 to 30 and 26 to 28 J/cm(2) for skin types I/II and III, respectively, were determined to be the maximal tolerated dose and resulted in severe erythema and crusting. The presence of severe erythema and discomfort precluded longer ALA exposure and higher fluence levels. No epithelial breakdown was observed at any treatment levels. CONCLUSION: The dose-response results defining the minimal erythema dose and the maximal tolerated dose of intense pulsed light-activated photodynamic therapy and radiofrequency energy have led to the development of 2 photorejuvenation protocols based on skin type and severity of photodamage. PMID- 15545531 TI - Nonanimal stabilized hyaluronic acid for lip augmentation and facial rhytid ablation. AB - OBJECTIVE: To evaluate the effectiveness of nonanimal stabilized hyaluronic acid as an injectable filling agent. DESIGN: Nonrandomized, retrospective, interventional case series. RESULTS: A total of 1446 consecutive patients (1029 women and 417 men) underwent intradermal injection of commercially available nonanimal stabilized hyaluronic acid (2242 treatments) for the enhancement of lip volume and contour and the reduction of visible facial rhytids. Almost 61% of all patients remained satisfied with their results after 9 months. The effect was longest in the glabellar and nasolabial fold areas. Minimal transient sequelae were noted. CONCLUSIONS: Nonanimal stabilized hyaluronic acid is an effective and safe facial soft tissue expander. Its duration varies with each facial area treated. PMID- 15545532 TI - Subcutaneous superficial musculoaponeurotic system grafting of the aging melolabial furrow. AB - OBJECTIVE: To describe a technique of subcutaneous superficial musculoaponeurotic system grafting during rhytidectomy to treat the aging melolabial furrow. DESIGN: Two hundred seventeen patients received bilateral superficial musculoaponeurotic system grafts from December 1, 1996, through June 30, 2003 (a 78-month period). The patients' charts were reviewed for demographic data, follow-up dates, and complications. Selected preoperative and postoperative photographs were viewed. RESULTS: In the 217 patients, 434 grafts were performed, with a mean follow-up of 23.2 months. Follow-up postoperative photographs showed effective long-term effacement of the melolabial furrow. There was a 0.9% complication rate involving 4 cases of unilateral graft infection that resolved with oral antibiotic therapy. CONCLUSIONS: Subcutaneous superficial musculoaponeurotic system grafting of the melolabial furrow is a safe, effective technique of treating the prominent melolabial furrow. It adds very little time to the rhytidectomy procedure, is well tolerated by patients, and shows durable results. PMID- 15545533 TI - Endoscopic forehead-lift using a bioabsorbable fixation device. AB - Endoscopic brow-lift is the method of upper face rejuvenation preferred by facial plastic surgeons and patients alike. The technique has undergone significant scrutiny regarding methods of fixation. Many techniques have been described, including external bolster dressings, tissue adhesives, Kirschner-wire fixation, cortical tunnels and troughs, and permanent or temporary screw and plate fixation. Many of these techniques are associated with postoperative alopecia, poor reliability, possible intracranial complications, and palpable hardware. Thus, the search for a safe, effective, reliable, and straightforward procedure continues. We describe our experience with an easy-to-insert bioabsorbable device that is fixed to the calvaria and provides multipoint fixation to the soft tissue. PMID- 15545534 TI - The use of custom-designed midfacial and submalar implants in the treatment of facial wasting syndrome. AB - Facial wasting syndrome is part of a lipodystrophy that occurs as a complication of highly active antiretroviral therapy. The loss of subcutaneous fat in the cheeks and temples results in a hollow-eyed, bony, emaciated appearance that is characteristic of the results of treatment of human immunodeficiency virus. Cessation of therapy results in a rebound in viral load and subsequent morbidity. The appearance of facial wasting syndrome is optimally treated with custom designed implants that are made using high-resolution computed tomography combined with surgeon input and computer-aided design and manufacturing technology. Twenty-two patients with facial wasting syndrome were treated using either submalar implants (in more moderate cases) or custom-designed implants (in more severe cases). In each patient, the appearance of volumetric soft tissue restoration was successfully achieved, returning a permanent and more healthful appearance to the face. PMID- 15545535 TI - Nonablative laser and light therapies for skin rejuvenation. AB - BACKGROUND: Multiple modalities have been described for skin rejuvenation, including ablative and nonablative therapies. Because of the prolonged recovery period associated with ablative procedures that injure the epidermis, nonablative skin treatments have grown increasingly popular. Various laser- and light-based systems have been designed or applied for promoting skin remodeling without damage to the epidermis. METHODS: Studies investigating the use of nonablative procedures for facial rhytids or acne scarring with clinical, histological, and objective quantitative measurements are systematically reviewed. RESULTS: Nonablative treatments are associated with clinical and objective improvements for the treatment of facial rhytids and acne scarring. Dermal remodeling seems to occur as a result of thermal injury, leading to dermal fibrosis without epidermal disruption. CONCLUSIONS: Although results are not as impressive as those of ablative treatments, nonablative procedures are effective in the treatment of photoaging and acne scarring. As technology in nonablative therapies continues to evolve, future laser and light sources may yield even more favorable results. PMID- 15545536 TI - Three-dimensional photography and its application to facial plastic surgery. PMID- 15545538 TI - Floyd Webb's Mme. Labaudt. PMID- 15545539 TI - UV-A1 phototherapy improves nephrogenic fibrosing dermopathy. PMID- 15545540 TI - Connective tissue remodeling induced by carbon dioxide laser resurfacing of photodamaged human skin. AB - OBJECTIVE: To quantitatively examine the dynamics of molecular alterations involved in dermal remodeling after carbon dioxide (CO(2)) laser resurfacing of photodamaged human skin. DESIGN: Serial in vivo biochemical analyses after laser therapy. SETTING: Academic referral center, Department of Dermatology, University of Michigan, Ann Arbor. Subjects Volunteer sample of 28 adults, 48 to 76 years old, with clinically evident photodamage of the forearms. Intervention Focal CO(2) laser resurfacing of photodamaged forearms and serial biopsies at baseline and various times after treatment. MAIN OUTCOME MEASURES: Reverse transcriptase real-time polymerase chain reaction technology and immunohistochemistry were used to assess levels of type I and type III procollagens; matrix metalloproteinases (MMPs) 1, 3, 9, and 13; tropoelastin; fibrillin; primary cytokines interleukin 1beta and tumor necrosis factor alpha; and profibrotic cytokine transforming growth factor beta1. RESULTS: Production of type I procollagen and type III procollagen messenger RNA peaked at 7.5 and 8.9 times baseline levels, respectively, 21 days after treatment and remained elevated for at least 6 months. Increases in messenger RNA levels of several cytokines (interleukin 1beta, tumor necrosis factor alpha, and transforming growth factor beta1) preceded and/or accompanied changes in collagen levels. Marked increases in messenger RNA levels of MMP-1 (39 130-fold), MMP-3 (1041-fold), MMP-9 (75-fold), and MMP-13 (767-fold) were noted. Levels of fibrillin and tropoelastin rose in a delayed fashion several weeks after treatment. CONCLUSIONS: The biochemical changes seen after CO(2) laser resurfacing proceed through a well-organized and highly reproducible wound healing response that results in marked alterations in dermal structure. These quantitative changes may serve as a means for comparison as other therapeutic modalities meant to improve the appearance of photodamaged skin are evaluated. PMID- 15545541 TI - Single-pass carbon dioxide laser skin resurfacing combined with cold-air cooling: efficacy and patient satisfaction of a prospective side-by-side study. AB - BACKGROUND: Ablative skin resurfacing with carbon dioxide (CO(2)) and erbium:YAG lasers is still considered the gold standard for treating rhytides, photodamage, and acne scars. However, the prolonged downtime and undesired concomitant effects that are involved have sent dermatologists looking for less invasive nonablative laser techniques to rejuvenate skin. OBJECTIVE: To combine cold-air cooling with single-pass CO(2) laser skin resurfacing to generate as much benefit as possible while minimizing the spectrum of adverse effects. STUDY DESIGN: The efficacy of single-pass CO(2) laser skin resurfacing on perioral and periorbital wrinkles was examined prospectively during a 6-month follow-up period. In a side-by-side comparison, the influence of simultaneous cold-air cooling on concomitant effects, pain tolerance, therapeutic success, and patient satisfaction was also studied. Eight patients with perioral and/or periorbital wrinkles underwent the procedure. During laser treatment, only the right half of each face was cooled using a cold-air system. RESULTS: Six months after treatment, a mild improvement of the wrinkles was observed in all cases. The use of cold-air cooling did not have any impact on the long-term results, although in a direct comparison between sides, it was observed that cooling reduced the recovery period from 3.9 +/- 1.5 (mean +/- SD) days to 3.5 +/- 1.4 days (P = .09) and helped postoperative erythema fade more quickly, from an average of 21.3 +/- 17.9 days to 11.7 +/- 3.9 days (P = .17). The reduction of pain was significant, which led to a much higher level of patient acceptance: on a numerical analog scale of 1 to 10, the rate decreased from an average of 6.8 +/- 1.8 (mean +/- SD) to 3.6 +/- 1.7 (P = .006). CONCLUSIONS: Given the clear decline in demand for invasive laser technologies, single-pass CO(2) laser skin resurfacing in conjunction with cold-air cooling is a worthwhile alternative both to conventional resurfacing and to conventional subsurfacing. The use of cold-air cooling not only minimizes intraoperative and postoperative adverse effects, it also contributes strongly to patient satisfaction. PMID- 15545542 TI - Treatment of atrophic facial acne scars with the 1064-nm Q-switched Nd:YAG laser: six-month follow-up study. AB - OBJECTIVES: To quantitatively assess improvement in acne scarring after a series of nonablative laser treatments and to determine efficacy at 1-, 3-, and 6-month follow-up after treatment. DESIGN: Before-after trial of consecutively selected patients. SETTING: Private practice at the Laser and Skin Surgery Center of New York, New York. PATIENTS: Eleven patients with mild to moderate atrophic acne scarring were treated. INTERVENTIONS: A 3-dimensional optical profiling imaging system was used to assess skin topography before, during, and after treatment. Patients were treated with a 1064-nm Q-switched Nd:YAG laser and reassessed after 3 treatment sessions and at 1, 3, and 6 months after the fifth treatment session. MAIN OUTCOME MEASURES: The skin roughness analysis was quantified at baseline and at each follow-up interval. Pain, erythema, and petechiae formation were assessed on 3-point scales. RESULTS: At midtreatment (1 month after the third treatment session), an 8.9% improvement in roughness analysis was seen. This improvement increased to 23.3%, 31.6%, and 39.2% at 1, 3, and 6 months after the fifth treatment, respectively. Patients reported mild to moderate pain with treatment. The only adverse effects noted were transient erythema and mild pinpoint petechiae. CONCLUSIONS: Treatment with the nonablative 1064-nm Q-switched Nd:YAG laser results in significant quantitative improvements in skin topography in patients with mild to moderate atrophic acne scars. Continued incremental improvements were noted at 1-, 3-, and 6-month follow-up, indicating ongoing dermal collagen remodeling after the treatment. PMID- 15545543 TI - Laser treatment of rosacea: a pathoetiological study. AB - OBJECTIVE: To study the effect of laser treatment on rosacea, a common facial skin disease with symptoms of blushing, redness, telangiectasis, papules, pustules, and diffuse swelling of the skin, we focused on the stinging sensation and performed immunohistochemical evaluation of nerve density and neuropeptide expression. DESIGN: Clinical investigation as well as the lactic acid (stinger) test was performed before and 3 months after the treatment with flashlamp pulsed dye laser, when skin biopsy specimens were also taken. SETTING: University hospital. PATIENTS: Thirty-two patients with rosacea, all with positive results from the lactic acid "stinger" test, were treated by flashlamp pulsed dye laser. MAIN OUTCOME MEASURES: The biopsy specimens were taken from the stinger-positive areas in the nasolabial folds, fixed in Lanas fixative (10% formalin and 0.4% picric acid), and analyzed for the expression of protein gene product 9.5 (general nerve marker), substance P, calcitonin gene-related peptide, and vasoactive intestinal polypeptide, using a biotinylated streptavidin technique. RESULTS: Thirty-one patients who were stinger positive before treatment showed decreased scores after treatment, and 1 patient had the same stinger test score before and after treatment. The number of protein gene product 9.5-positive fibers in the epidermis (P< .05) as well as the papillary dermis (P< .01) was decreased. This was also the case for substance P in the papillary dermis (P< .001), whereas no evident difference was noted for vasoactive intestinal polypeptide and calcitonin gene-related peptide. No difference was found for contact between nerves and vessels (factor VIII positive). CONCLUSIONS: Laser treatment of rosacea that destroys small vessels has a good medical relevance because it reduces the unpleasant symptoms of the sensitive skin. A neurogenic etiology of stinging may be possible. PMID- 15545544 TI - Effect of volume and concentration on the diffusion of botulinum exotoxin A. AB - OBJECTIVE: To investigate whether the volume of solution used to inject equivalent units of botulinum exotoxin A affects the diffusion of toxin and areas of rhytid diminution in the treatment of dynamic forehead lines. DESIGN: Ten volunteers with dynamic forehead lines were included. Each study patient received a single injection at a point 2.5 cm above the orbital rim on either side of the forehead with equivalent units, but in different volumes, of botulinum exotoxin A. The sides of injection were randomized; one side of the patient's forehead was injected first with 5 U of botulinum exotoxin A in 0.25 mL (2 U/0.1 mL) of preserved saline in the midpupillary line, followed by injection of the other side with 5 U in 0.05 mL of preserved saline (2 U/0.02 mL). There was a 5-fold difference in volume injected. Subjects were evaluated 14 days later for total area affected during visual inspection of the subjects' foreheads during active muscle contraction. SETTING: Private dermatology office. MAIN OUTCOME MEASURE: Visual inspection to measure the area of rhytid effacement in both height and width. RESULTS: The area affected by the botulinum exotoxin A injection was 50% greater in the side with the larger volume in 9 of 10 subjects. The average area affected was 6.05 cm(2) for the injection of the larger volume compared with 4.12 cm(2) for the injection with the smaller volume. The shape of rhytid effacement was oval, rather than round, with the average width longer than the average height. CONCLUSIONS: In this prospective, randomized, controlled study, we found that injection of botulinum exotoxin A in low concentration and higher volume resulted in greater diffusion and a larger affected area. The pattern of toxin spread is altered by muscular contraction in the injected sites. These results show that the dilution has implications on the desired effect of botulinum exotoxin A. PMID- 15545545 TI - An education theory-based method to teach a procedural skill. AB - OBJECTIVE: To determine the effectiveness of an education theory-based method to teach students to place and tie a simple interrupted stitch. DESIGN: A teaching intervention before-after trial. SETTING: Dermatology department, academic university. PARTICIPANTS: Fourth-year medical students and dermatology residents. MAIN OUTCOME MEASURES: Scores on a 12-criterion grading instrument before and after instruction. RESULTS: The scores for medical students and residents in each class showed significant improvement. The mean score for all participants (N = 23) rose by 24% after instruction (P< .001). Scores in 9 of the 12 graded performance areas improved significantly after instruction, including scores in tissue damage/teeth marks (P<.001), needle dulled/bent (P< .001), needle loaded properly and knots square (P = .01), throws done correctly (P = .01), stitch tension and needle entry/exit angle (P = .02), amount of suture used (P = .03), and correct number of throws (P = .04). In addition, participants' confidence increased significantly after instruction (P<.001). No difference was noted between men and women in preinstruction vs postinstruction score improvement. CONCLUSIONS: This teaching method can be effectively used to teach students to place and tie a simple interrupted stitch. Once validated and expanded, it may prove useful in shortening and standardizing procedural skill training and in objectively documenting competency. PMID- 15545546 TI - Teaching and evaluation of surgical skills in dermatology: results of a survey. AB - OBJECTIVES: To assess how the surgical skills of residents are taught and evaluated within dermatology residency programs in the United States; to assess which surgical techniques training directors and residents consider important for residents to perform or at least understand by the end of residency training. METHODS: A 126-question survey was sent to all 106 of the US dermatology residency programs accredited by the Accreditation Council for Graduate Medical Education. Contact was initially made via e-mail. Surveys were addressed to the program director, surgical training director, and chief resident of each program. A follow-up survey was mailed to nonresponders. RESULTS: Ninety-five surveys were returned representing 71 (67%) of 106 programs. Eighty-nine percent of programs (n=63) reported having a formal curriculum in dermatologic surgery. Among programs represented, 97% (n=69) taught surgical skills in the procedure room, 84% (n=57) used pigs' feet, and fewer than 10% (n=6) used human cadavers. Ninety four percent of programs (n=61) scheduled surgical lectures; two thirds (n=41) formally assigned surgical reading, and over half (n=36) used Web-based lectures to teach skills. To assess training, most programs (86%; n=50) used subjective global evaluation at the end of a surgery rotation. Fewer than 30% (n=15) discussed specific objectives prior to the rotation. Only about 25% of programs (n=17) reported the use of written or oral examinations to assess resident surgery skills. Traditional biopsy and simple surgical procedures were reported as most important to know and perform. Interest by both faculty members and residents in more advanced surgical techniques was more limited and variable. Cosmetic surgery techniques were most likely to be viewed as unimportant. CONCLUSIONS: Most dermatology programs teach surgical skills by traditional apprenticeship methods supplemented by work in pigs' feet laboratory classes and regularly scheduled lectures. Skill assessment is mainly done through subjective means. Almost all respondents thought that basic biopsy and excisional skills were essential for residents to know and perform. More complex surgical techniques and the use of lasers were considered less important. Cosmetic techniques were those most frequently viewed as unimportant. PMID- 15545547 TI - Nonablative facial remodeling: erythema reduction and histologic evidence of new collagen formation using a 300-microsecond 1064-nm Nd:YAG laser. AB - BACKGROUND: A variety of nonablative lasers have been used to improve skin color and toning. Evidence of new collagen has been seen. Using blinded observer analysis of electron microscopic changes, we have documented the effect of a nonablative Nd:YAG laser on collagen production and its relationship to patient age. OBSERVATIONS: Ultrastructural analysis of 9 patients showed a decrease in overall collagen fiber diameter in the papillary dermis at 1 month and 3 months after 3 treatment sessions. This is consistent with the formation of new collagen. Younger patients had a greater decrease in collagen fiber diameter compared with older patients. The change in collagen fiber diameter with time as well as the relationship between that change and the patient's age were statistically significant (P<.001). Photographic evaluation showed that those patients with preexisting erythema showed improvement in erythema along with an associated improvement in skin quality. There were no adverse events. CONCLUSIONS: Microsecond Nd:YAG lasers appear to be safe for nonablative laser remodeling. Our study indicates that microsecond Nd:YAG lasers can produce new collagen formation in the papillary dermis. In addition, the condition of patients with erythema may be improved. Younger patients may form more new collagen compared with older patients with photodamage. PMID- 15545548 TI - The safety of office-based surgery: review of recent literature from several disciplines. AB - OBJECTIVE: To review recent literature pertaining to adverse outcomes and mortality associated with office-based surgery. STUDY SELECTION: Representative articles from the general and plastic surgery, medical, health regulatory, and dermatology literature. DATA EXTRACTION: Information regarding which surgical treatments should be performed, which specialties should perform them, what level of anesthesia is appropriate, and who should administer it was assessed, with particular attention to issues of patient safety. CONCLUSIONS: Office-based surgery is safe and cost-effective. We caution against attempts to prohibit or severely restrict this important aspect of medical care. PMID- 15545549 TI - Rejuvenating the aging face. PMID- 15545550 TI - Beyond the biopsy in laser surgery. PMID- 15545552 TI - A painful blue nodule on the dorsum of the hand. PMID- 15545553 TI - Pustular plaques on the face of a patient with renal failure. PMID- 15545554 TI - A slowly growing ulcerated nodule on the scalp. PMID- 15545555 TI - A pigmented verrucous plaque on the cheek. PMID- 15545556 TI - Nephrogenic fibrosing dermopathy: are ACE inhibitors the missing link? PMID- 15545557 TI - Dermoscopy of disseminated superficial actinic porokeratosis. PMID- 15545559 TI - Comparison of a new fibrin sealant with standard topical hemostatic agents. AB - BACKGROUND: Bleeding following liver resection continues to be a significant morbidity of the procedure. Fibrin sealants represent an improvement over conventional topical hemostatic agents, because they contain components that actively form clot. However, most available agents contain nonhuman protein, which represents an immunologic risk. HYPOTHESIS: An investigational surgical fibrin sealant (Crosseal; American Red Cross, Washington, DC) composed of human clottable proteins and human thrombin is more effective than standard topical hemostatic agents in reducing the time required to achieve hemostasis after liver resection. DESIGN: Prospective, randomized, controlled trial. SETTING: Fifteen major referral centers in the United States and the United Kingdom. METHODS: After liver resection using standard surgical techniques, 121 patients seen between May 1999 and May 2000 were randomized to treatment with a 2-component fibrin sealant (n=58) or to standard topical hemostatic agents, used singly or in combination (n=63). Up to 10 mL of Crosseal was administered by a spray applicator, as recommended by the manufacturer, whereas agents used in the control group were applied according to their instructions for use. MAIN OUTCOME MEASURES: The primary outcome measured was time to hemostasis. Secondary outcomes measured included blood loss between application of the hemostatic agent and closure of the abdomen, duration of postoperative biliary drainage, and the occurrence of complications, defined a priori as reoperation for any reason, development of abdominal fluid collections, or bilious appearance of drained fluid for at least 1 day postoperatively. RESULTS: The mean time to hemostasis was 282 seconds with Crosseal, compared with 468 seconds with standard agents (2 sided; P =.06), for the 116 efficacy-evaluable patients. Hemostasis was achieved within 10 minutes in 53 patients (91.4%) treated with the study fibrin sealant and in 44 control patients (69.8%) (2-sided; P =.003). Intraoperative blood loss was similar in the 2 groups. In the Crosseal group, the percentage of patients developing postoperative complications was 17.2%, compared with 36.5% in the control group (2-sided; P =.02). CONCLUSIONS: Compared with the use of standard topical hemostatic agents, Crosseal fibrin sealant significantly reduced the time to achieve hemostasis following liver resection. Patients treated with the new fibrin sealant also experienced significantly fewer postoperative complications. PMID- 15545560 TI - Multicenter, prospective, double-blind, randomized trial of laparoscopic nissen vs anterior 90 degrees partial fundoplication. AB - HYPOTHESIS: Laparoscopic anterior 90 degrees partial fundoplication for gastroesophageal reflux is associated with a lower incidence of postoperative dysphagia and other adverse effects compared with laparoscopic Nissen fundoplication. DESIGN: A multicenter, prospective, double-blind, randomized controlled trial. SETTING: Nine university teaching hospitals in 6 major cities in Australia and New Zealand. PARTICIPANTS: One hundred twelve patients with proven gastroesophageal reflux disease presenting for laparoscopic fundoplication were randomized to undergo either a Nissen (52 patients) or an anterior 90 degrees partial procedure (60 patients). Patients with esophageal motility disorders, patients requiring a concurrent abdominal procedure, and patients who had undergone previous antireflux surgery were excluded from this study. INTERVENTIONS: Laparoscopic Nissen fundoplication with division of the short gastric vessels or laparoscopic anterior 90 degrees partial fundoplication. MAIN OUTCOME MEASURES: Independent assessment of dysphagia, heartburn, and overall satisfaction 1, 3, and 6 months after surgery using multiple clinical grading systems. Objective measurement of esophageal manometric parameters, esophageal acid exposure, and endoscopic assessment. RESULTS: Postoperative dysphagia, and wind-related adverse effects were less common after a laparoscopic anterior 90 degrees partial fundoplication. Relief of heartburn was better following laparoscopic Nissen fundoplication. Overall satisfaction was better after anterior 90 degrees partial fundoplication. Lower esophageal sphincter pressure, acid exposure, and endoscopy findings were similar for both procedures. CONCLUSIONS: At the 6-month follow-up, laparoscopic anterior 90 degrees culine partial fundoplication is followed by fewer adverse effects than laparoscopic Nissen fundoplication with full fundal mobilization, and it achieves a higher rate of satisfaction with the overall outcome. However, this is offset to some extent by a greater likelihood of recurrent gastroesophageal reflux symptoms. PMID- 15545561 TI - The clinical risk score: emerging as a reliable preoperative prognostic index in hepatectomy for colorectal metastases. AB - HYPOTHESIS: The purpose of this study was to examine the validity of the clinical risk score (CRS), a prognostic tool developed by Fong et al, when translated to another center. DESIGN: This study assesses 5 independent preoperative prognostic criteria, nodal status of the primary lesion, disease-free interval, number of hepatic metastases, size of the largest metastasis, and preoperative carcinoembryonic antigen level, to determine a preoperative CRS for each patient included in the study. SETTING: The hepatobiliary unit of The Queen Elizabeth Hospital, Adelaide, South Australia. PATIENTS: Medical records of patients admitted to The Queen Elizabeth Hospital undergoing potentially curative hepatic resection for colorectal metastases during the period of July 1993 to April 2003 were included in the study. Main Outcome Measure The primary outcome measure of the study was survival. The calculated CRS was analyzed with respect to patient postoperative survival. RESULTS: During the 10-year period, 77 patients underwent hepatic resection. Overall survival rates for 1, 3, and 5 years were found to be 80.9%, 57.5%, and 42.3%, respectively. One- and 5-year survival rates for CRSs of 0 and 1 were found to be 93.8% and 72.5%, respectively; for scores of 2 and 3, 76.6% and 31.2%, respectively;and for scores of 4 and 5, 75% and 0%, respectively. No patient with a CRS greater than 3 survived more than 2 years. CONCLUSION: This study validates the CRS, finding it to be highly predictive of patient outcome and survival. PMID- 15545562 TI - Clinicopathologic and radiopharmacokinetic factors affecting gamma probe-guided parathyroidectomy. AB - HYPOTHESIS: The kinetics of technetium Tc 99m sestamibi (MIBI) in primary hyperparathyroidism are variable and affected by the cellular size of the abnormal glands, the parathyroid hormone levels, and the functional expression of P-glycoprotein (Pgp). The success of gamma probe-guided parathyroidectomy is closely related to the parathyroid-to-thyroid activity ratio at the time of surgery. Preoperative determination of maximum uptake ratio may improve the surgical outcome. DESIGN: Thirty-one patients with primary hyperparathyroidism attributed to a solitary parathyroid adenoma (27 patients) or multiglandular hyperplasia (4 patients) underwent dynamic MIBI imaging preoperatively. Maximum MIBI activity and activity elimination half-life in the abnormal parathyroid glands and thyroid glands were measured, and the maximum uptake ratio was calculated. After a second MIBI injection on the day of surgery, all patients underwent gamma probe-guided parathyroidectomy and cervical exploration. Timing of surgery after MIBI injection was individualized according to the optimal time to surgery (time to maximum uptake ratio), which was determined by preoperative scintigraphy. During surgery, the gamma probe was used to measure ex vivo counts of excised lesions and adjacent postexcision normal tissue (background). Image characteristics, MIBI kinetics, and gamma probe findings were correlated with gland volume, oxyphil cell content, Pgp expression, and serum parathyroid hormone levels. RESULTS: Probe localization of abnormal glands at maximum uptake ratio was successful in all patients. The volume of the parathyroid lesion ranged from 0.03 to 9.8 mL (median, 0.7 mL). Parathyroid maximum MIBI activity correlated with the volume of the gland (r = 0.54, P = .002) and serum parathyroid hormone level (r = 0.58, P = .001). No correlation between maximum MIBI activity and oxyphil cell content or Pgp expression could be demonstrated. Elimination half life of MIBI from parathyroid inversely correlated with Pgp (r = -0.36, P = .05). The ex vivo lesion-background count ratio positively correlated with volume of the gland (r = 0.66, P = .001) and parathyroid hormone level (r = 0.48, P = .006). Ex vivo lesion counts and Pgp expression were negatively correlated (r = 0.37, P = .04). CONCLUSIONS: A strong relationship between volume of the parathyroid gland, serum parathyroid hormone levels, and MIBI uptake exists in primary hyperparathyroidism. Gamma probe-guided localization of abnormal gland(s) can be more successful if surgery is undertaken at maximum uptake ratio. High Pgp expression increases MIBI parathyroid clearance rate, decreases gamma probe counts, and may significantly alter the optimal time to surgery. PMID- 15545563 TI - 1% lymphazurin vs 10% fluorescein for sentinel node mapping in colorectal tumors. AB - HYPOTHESIS: Ten percent fluorescein may be successfully used as an alternative to 1% Lymphazurin (1% isosulfan blue; US Surgical Corp, North Haven, Conn) in sentinel lymph node (SLN) mapping for the accurate staging of colorectal tumors. DESIGN: Review of prospectively gathered data. SETTING: University-affiliated regional medical center. PATIENTS: Sentinel lymph node mapping was performed in 120 consecutive patients with colorectal malignancies. INTERVENTIONS: The first 1 to 4 blue nodes detected within 5 minutes were designated as Lymphazurin-detected SLNs. The first 1 to 4 fluorescent nodes seen under the Wood light were designated as fluorescein-detected SLNs. Multilevel serial sections for hematoxylin-eosin and immunohistochemistry studies for cytokeratin were performed on all SLNs. MAIN OUTCOME MEASURES: Successful mapping, accuracy, skip metastasis, adverse reactions, occult micrometastases detection, and cost. RESULTS: Mapping was successful using Lymphazurin in 99% of the patients vs 97% of the patients using fluorescein (P =.89). The accuracy of predicting nodal metastases with each tracer was 95.8% vs 93.1%, respectively (P =.82). The skip metastases rate was 4.2% for Lymphazurin vs 6.9% for fluorescein (P =.37). The 5 patients in whom nodal disease was only identified as occult micrometastasis in the SLNs had a total of 5 SLNs, all of which were identified by both tracers. No adverse reactions occurred. The cost for Lymphazurin was $99.00, while the cost for fluorescein was $2.10. CONCLUSIONS: With the exception of cost, there were no statistically significant differences between the 2 dyes. While easy availability and lower cost remain distinct advantages of fluorescein, Lymphazurin remains the gold standard. In patients with known hypersensitivity to Lymphazurin and when availability and cost are an issue, fluorescein may be used effectively for SLN mapping in colorectal tumors. PMID- 15545564 TI - Complication of thyroidectomy in patients with radiation-induced thyroid neoplasms. AB - HYPOTHESIS: The complication rate for thyroidectomy is the same in patients with and without a history of radiation exposure. DESIGN: Retrospective medical record review of 171 consecutive patients who had a previous history of radiation treatment and had undergone a thyroid operation from 1961 to 1999. SETTING: University of California, San Francisco, Medical Center and affiliated hospitals. PATIENTS: We selected 107 patients with a history of radiation exposure who had undergone thyroid operations (81 total thyroidectomies) and 107 control patients who underwent comparable operations but had no history of radiation exposure. RESULTS: Among patients with a history of radiation exposure (mean age, 47.2 years), there was 1 recurrent nerve injury, 1 external nerve injury, 20 patients with transient hypocalcemia, and 1 patient with a hematoma. Among patients without a history of radiation exposure (mean age, 47.5 years), there were 2 recurrent nerve injuries, 18 patients with transient hypocalcemia, and 1 patient with a hematoma. All cases of hypocalcemia and recurrent nerve injury in both groups were transient. One patient had a permanent superior laryngeal nerve injury. In patients who underwent operations since January 1, 1990, duration of hospitalization was 1.2 days in patients with a history of radiation exposure (65 patients) and 1.1 days in patients without (101 patients). CONCLUSIONS: Our data document that the risk of transient and permanent complications after total thyroidectomy is similar in patients with and without a history of radiation exposure. The relatively low long-term complication rate supports prophylactic total thyroidectomy for patients with thyroid nodules and a history of radiation exposure. PMID- 15545565 TI - Simultaneous liver-kidney transplantation for adult recipients with irreversible end-stage renal disease. AB - HYPOTHESIS: Combined liver-kidney transplantation is safe (low morbidity and acceptable mortality) and effective in patients with end-stage liver disease. Although refinements in surgical technique have resulted in better patient and allograft outcomes, the negative impact of renal insufficiency on survival in patients undergoing liver transplantation has been widely reported, although some aspects are controversial. DESIGN: Analysis of the clinical characteristics and outcome in the management of patients undergoing combined liver-kidney transplantation. The end points were operative mortality, morbidity, and long term survival. SETTING: University Hospital 12 de Octubre. PATIENTS: Between May 1986 and December 2001, 820 liver transplantations were performed. There were 16 cases (1.96%) of combined liver-kidney transplantations, which represent the sample of this study. RESULTS: Mean +/- SD follow-up of 42.2 +/- 29 months: 6 patients died (37.5% mortality rate). There were 4 (25%) hospital deaths within 6 months following surgery and 2 after 6 months (4 sepsis, 1 refractory heart failure, and 1 recurrent hepatitis C virus disease). Univariate analysis related to mortality included age, sex, etiology, preoperative creatinine level, United Network for Organ Sharing status, Child-Pugh score, type of hepatectomy (piggyback), intraoperative blood product administration, and the presence of postoperative complications. The only 2 significant factors were the presence of postoperative complications (P = .01) and the United Network for Organ Sharing status (P = .02). Crude survival rate was 62.5%. Actuarial survival rates were 80%, 71%, and 60% at 1, 3, and 5 years, respectively. CONCLUSION: Because end stage renal disease is not a formal contraindication for liver transplantation, a combined liver-kidney transplantation for adults with end-stage renal disease can be done safely and effectively. PMID- 15545566 TI - Improving team structure and communication: a key to hospital efficiency. AB - HYPOTHESIS: Improving team structure and heightening communication will help provide cost-effective and high-quality patient care for general surgery patients. DESIGN: This study surveys teamwork initiatives and their effects on specific variables related to patient care. PATIENTS: The study population comprised all patients admitted to the hospital's general surgery teams during 5 years 3 months. SETTING: Tertiary care hospital. INTERVENTIONS: A complete restructuring of the patient care team for general surgery patients admitted to the hospital. The intervention occurred midway through the study period. MAIN OUTCOME MEASURES: Mean length of stay for general surgery patients as a marker of team efficiency and a standardized patient satisfaction survey. RESULTS: The mean length of stay after initiation of the restructured care team was significantly shorter than before initiation. The significance was present despite a consistent patient acuity measure and was associated with a high patient satisfaction level. CONCLUSIONS: Restructuring the patient care team yielded a decreased mean length of stay while maintaining a high level of patient satisfaction. This analysis helps validate a hospital-wide initiative to maintain a high level of patient care while increasing patient volume. PMID- 15545567 TI - Hypothyroidism and adrenal insufficiency in sepsis and hemorrhagic shock. AB - HYPOTHESIS: We hypothesized that hypothyroidism and adrenal insufficiency frequently occur together in critically ill patients. DESIGN: A prospective observational study. SETTING: Surgical intensive care unit of a university affiliated tertiary referral center. PATIENTS: Sixty-six consecutive patients with severe sepsis, septic shock, and hemorrhagic shock who required pulmonary artery catheterization for resuscitation were studied. INTERVENTIONS: Thyrotropin and baseline cortisol levels were obtained at 3 am followed by intravenous injection of 250 mug of cosyntropin, a synthetic adrenocorticotropic hormone derivative. A second measurement of the cortisol level was performed 1 hour later. MAIN OUTCOME MEASURES: Incidence of hypothyroidism and adrenal insufficiency and mortality. RESULTS: Mean (SD) age was 62 (19) years. The mean (SD) Acute Physiology and Chronic Health Evaluation II score was 21 (5). Twenty seven patients (40.9%) had severe sepsis, 31 (46.9%) had septic shock, and 8 (12.1%) had hemorrhagic shock. Five patients (7.6%) had hypothyroidism alone and 35 (53.0%) had only adrenal insufficiency. Eight patients (12.1%) had both hypothyroidism and adrenal insufficiency. All patients with endocrine abnormalities were treated. Mortality for the total group was 15 (22.7%) of 66 patients. CONCLUSION: There is a 12% incidence of simultaneous hypothyroidism and adrenal insufficiency in our study and the routine testing for both may be indicated in this population of critically ill patients. PMID- 15545568 TI - Professional satisfaction of women in surgery: results of a national study. AB - HYPOTHESIS: Individual, group, and organizational factors influence the professional satisfaction of women surgeons in Austria. DESIGN: Survey on professional and private issues sent out by mail in 2000 and 2001. SETTING: Women surgeons working in hospitals and/or in private practices and those who were retired or on maternity leave. PARTICIPANTS: All 351 Austrian women surgeons of all core surgical specialties (general, trauma, pediatric, plastic, thoracic, and cardiovascular), certified or in training, were addressed. MAIN OUTCOME MEASURES: Proportional odds regression models were used to correlate professional satisfaction with objectively measurable prognostic factors such as age, surgical subspecialty, status of training, type of hospital, location of work (federal states vs the capital), status of activity (active vs on maternity leave), profession of private partner, number of children, and subjectively assessed prognostic factors such as operative volume and departmental organization. RESULTS: The response rate was 58.7% (206/351). One hundred eighty-seven surgeons active or on maternity leave-were included in the analysis. Higher satisfaction was reported by active surgeons in subspecialties, certified surgeons, comparatively younger and older surgeons, surgeons working in hospitals outside the capital, and surgeons with a physician as a partner. When entering subjectively assessed variables into the model, the quality of departmental organization and operative volume (P<.001), as well as the status of activity (P<.001), had the strongest effect. CONCLUSIONS: Women surgeons' professional satisfaction highly depends on departmental organization and status of activity. Inadequate leadership, low operative volume, and being on maternity leave have a negative effect on job satisfaction. Private factors seem to be of little influence. Optimal departmental organization would help women to reconcile their professional and their private lives. PMID- 15545569 TI - Identification and categorization of technical errors by Observational Clinical Human Reliability Assessment (OCHRA) during laparoscopic cholecystectomy. AB - HYPOTHESIS: Surgical operative performance benefits from analysis of the mechanisms underlying technical errors committed during surgery. DESIGN: Prospective study using the Observational Clinical Human Reliability Assessment (OCHRA) system and complete unedited videotapes of the operations. SETTING: Three National Health Service hospitals within the United Kingdom. PATIENTS: Two hundred consecutive patients with symptomatic gallstone disease. INTERVENTIONS: Elective laparoscopic cholecystectomy for symptomatic gallstone disease by surgeons, who were blind to the nature and objectives of the study, using their usual operative technique. MAIN OUTCOME MEASURES: Surgical consequential and inconsequential operative errors. RESULTS: The analysis of 38 062 steps of the 200 laparoscopic cholecystectomies performed by 26 surgeons identified 2242 errors. The mean +/- SD total, inconsequential, and consequential errors per surgical procedure were 11.0 +/- 8.0, 8.0 +/- 6.0, and 4.0 +/- 3.0, respectively. Dissection of the Calot triangle (second task zone of the operation) incurred more total errors (6.5 +/- 5.4) compared with the first (2.9 +/- 2.8, P<.001) and third (5.1 +/- 3.9, P<.05) task zones. This translated to a higher error probability (6.9% vs 3.5% for the first and 5.5% for third task zones). The combined sharp and blunt dissection method had fewer errors than the blunt/teasing dissection technique (9.45 +/- 7.6 vs 13.9 +/- 7.3, P<.001) although different surgeons were involved. The most serious consequences were encountered during dissection with the electrosurgical hook knife. CONCLUSION: This study has confirmed that the Observational Clinical Human Reliability Assessment system provides a comprehensive objective assessment of the quality of surgical operative performance by documenting the errors, the stage of the operation in which errors are enacted most frequently, and where these errors have serious consequences (hazard zones). PMID- 15545570 TI - Impact of primary resection on the outcome of patients with perforated diverticulitis. AB - BACKGROUND: Primary resection has replaced the conventional drainage procedure in the management of patients with generalized peritonitis complicating diverticular disease of the colon. This study investigates the impact of primary resection on operative mortality, identifies predictors of mortality, and compares the results with those of our earlier experience. HYPOTHESIS: Primary resection of the perforated diseased segment of the colon is associated with lower mortality rates than the drainage procedure in patients with Hinchey stages 3 and 4 diverticulitis. DESIGN: Retrospective analysis. SETTING: Tertiary care referral center. PATIENTS: We included 138 consecutive patients who underwent emergent operation for generalized peritonitis complicating diverticular disease of the colon (Hinchey stages 3 and 4) during a period of 16 years (January 1983 to May 1999). MAIN OUTCOME MEASURES: The 30-day mortality rate was analyzed and predictors of mortality identified. RESULTS: Patients were classified as having spreading purulent peritonitis (n = 44, 31.9%), diffuse peritonitis (n = 64, 46.4%), or fecal peritonitis (n = 30, 21.7%). One hundred thirty-one patients (94.9%) underwent primary resection, 6 patients (4.3%) underwent resection and primary anastomosis, and 1 patient required total colectomy and end ileostomy. Thirteen of the 138 patients in the present group died (1983-1998), representing a perioperative mortality rate of 9%. There was no significant difference in mortality when compared with our earlier study (1972-1982), which had a mortality rate of 12%, considering that more than 25% of the patients in that group were managed by colostomy and drainage alone. Factors identified univariately as predictors of mortality were age of more than 70 years (P = .047), 2 or more comorbid conditions (P<.01), obstipation at initial examination (P = .02), use of steroids (P = .01), and perioperative sepsis (P<.001). CONCLUSIONS: Primary resection has become the standard practice for patients with generalized peritonitis complicating diverticulitis. Mortality rates have not significantly declined despite more aggressive surgical management of the septic source. Because advanced age, comorbid conditions, and perioperative sepsis predict mortality, it is suggested that further reduction in mortality will require improvement in medical management of perioperative sepsis and comorbid conditions. PMID- 15545571 TI - Increased survival among severe trauma patients: the impact of a national trauma system. AB - HYPOTHESIS: The survival of severe trauma patients is affected by the implementation of a national trauma system, which brought about developments both at the hospital and prehospital levels during the past decade. DESIGN: A retrospective cohort study of all severely injured patients (Injury Severity Score >16) recorded in the Israeli National Trauma Registry at all level I trauma centers in Israel from January 1, 1997, to December 31, 2001. Inpatient death rates were examined overall and by subgroups. SETTING: The National Trauma Registry includes trauma (International Statistical Classification of Diseases, 9th Revision, Clinical Modification diagnosis codes 800-959) hospitalizations, patients who were transferred to or from other hospitals, and those who died in the emergency department. It excludes patients who were dead on arrival, discharged following treatment in the emergency department, and patients who do not fall into the definition of trauma. Main Outcome Measure Inpatient death. RESULTS: Seven thousand four hundred twenty-three severe trauma patients were recorded. Inpatient death rates decreased significantly from 21.6% in 1997 to 14.7% in 2001. The odds ratios of mortality in 1998 through 2001 vs 1997, adjusted for year, age, sex, penetrating injury, and severity of injury (Injury Severity Score >25), were 0.92, 0.89, 0.70, and 0.65, respectively, confirming the downward trend. CONCLUSIONS: A steady significant reduction in the inpatient death rate of severe trauma patients hospitalized at all level I trauma centers in Israel between 1997 and 2001 was observed. Although a single factor that explains the reduction was not identified, it is evident that the establishment of the trauma system brought about a significant decrease in mortality. We believe that integrated cooperation of various components of the national trauma system in Israel across the years may explain the reduction. PMID- 15545572 TI - Isolated limb perfusion for unresectable melanoma of the extremities. AB - HYPOTHESIS: In patients with truly unresectable melanoma of the extremities, results after isolated limb perfusion (ILP) are absent in the literature. Complete response rates are probably lower than the reported 54% for locoregional recurrent melanoma. In these patients, ILP with melphalan and tumor necrosis factor alpha (TNF-alpha) could be superior to ILP with melphalan alone. DESIGN: Retrospective analysis with a median follow-up period of 21 months (interquartile range, 9-40 months). SETTING: Two tertiary care cancer centers in the Netherlands. PATIENTS: We assessed all 130 consecutive patients who underwent ILP for unresectable melanoma of the extremities, performed between 1978 and 2001. Of these patients, 38% had stage IIIA melanoma and 45% had stage IIIAB melanoma according to criteria of the MD Anderson Cancer Center. Lesions were considered unresectable on the basis of their size, number, or localization. INTERVENTIONS: Forty ILPs were performed with melphalan, and 90 were done with TNF-alpha and melphalan. MAIN OUTCOME MEASURES: Response rate, disease-free survival, limb salvage rate, and overall survival. RESULTS: In 45% of the patients, a complete response was attained after ILP with melphalan (95% confidence interval, 29%-61%) compared with 59% after ILP with TNF-alpha and melphalan (95% confidence interval, 49%-69%; P = .14). The time to complete response was 3 months (interquartile range, 2-6 months) vs 2 months (interquartile range, 1-3 months; P = .01), respectively. The recurrence rate and median limb recurrence-free survival were not significantly different for both ILP types. The overall limb salvage rate was 96%. Overall 5-year survival was 29% (95% confidence interval, 20%-38%). The ILP type was not an independent prognostic factor for complete response, nor was limb recurrence-free survival, whereas stage IIIA was a favorable prognostic factor (P = .01 and P = .02, respectively). Favorable prognostic factors for improved survival were complete response (P<.001) and a tumor size of 3 cm or less (P = .01). CONCLUSIONS: In more than half of the patients with truly unresectable melanoma of the extremities, a complete response was obtained after ILP with melphalan with or without TNF-alpha. The ILP type was not an independent prognostic factor for complete response, limb recurrence-free survival, or overall survival. PMID- 15545573 TI - Performing laparoscopic adrenalectomy safely. PMID- 15545574 TI - Trocar site hernia. AB - OBJECTIVE: To review the relationship between the pathogenesis and clinical manifestations of trocar site hernias seeking to confirm the definition of trocar site hernias by classification. DATA SOURCES: We searched this subject in English on MEDLINE by combining the words "trocar," "port," "hernia, and "laparoscopy."Data Extraction and STUDY SELECTION: We limited the main operations to cholecystectomy, colon and rectal surgery, fundoplication, and gastric surgery; finding 44 reports on these procedures. Of these, 19 were case reports, 18 were original articles (setting criteria; the incidence of the trocar site hernia was clarified, and involved >100 patients), and 7 technical notes on "how to do it" were collected. We obtained 19 additional reports using the references of those previously obtained. We, thus, reviewed 63 reports (24 case reports, 27 original articles, 7 technical notes, and 5 review articles). DATA SYNTHESIS: Trocar site hernia was classified into 3 types. The early-onset type that occurred immediately after the operation, with a small-bowel obstruction, especially the Richter hernia, frequently developing. The late-onset type that occurred several months after the operation, mostly with local abdominal bulging with no small-bowel obstruction developing. The special type that occurred indicated the protrusion of the intestine and/or omentum. Trocar site hernias with fascial defects of 10 mm or larger should be closed, including the peritoneum. Opinion varied if a 5-mm trocar site defect should be closed. CONCLUSION: It is useful to clearly classify trocar site hernias to improve management of laparoscopic procedures. PMID- 15545575 TI - Image of the month. Cerebral fat embolism. PMID- 15545576 TI - When living related organ donation does not entail a healthy organ: moral implications for both the donor and the recipient. PMID- 15545577 TI - A little goes a long way in tinnitus research. PMID- 15545578 TI - Serum cotinine level and incident hearing loss: a case-control study. AB - BACKGROUND: A growing body of literature suggests an association between cigarette smoking and hearing loss. OBJECTIVE: To assess the relation between levels of serum cotinine, a biomarker of exposure to tobacco smoke, and incident hearing loss. DESIGN: A cross-sectional, incident, case-control study of participants selected from a population-based cohort. SETTING: Testing was conducted at the Beaver Dam Community Hospital, Beaver Dam, Wis. PARTICIPANTS: A total of 197 participants with incident hearing loss and 394 control participants, aged 53 to 75 years, selected from the 2800 participants of the 5 year follow-up examination of the population-based Epidemiology of Hearing Loss Study, 1998-2000. MAIN OUTCOME MEASURE: Incident hearing loss. The incidence of hearing loss was defined as a pure-tone average of thresholds at 500, 1000, 2000, and 4000 Hz greater than 25-dB hearing level in either ear at follow-up among those without hearing loss at baseline. RESULTS: No significant associations were found between serum cotinine levels and incident hearing loss. CONCLUSIONS: These results were not consistent with a previous report, which found cross-sectional associations between prevalent hearing loss and current smoking and environmental tobacco smoke exposure in the home. Longer-term longitudinal studies of smoking and/or serum cotinine levels and the subsequent development of hearing loss may help clarify these associations. PMID- 15545579 TI - Auditory effects after organ preservation protocol for laryngeal/hypopharyngeal carcinomas. AB - OBJECTIVE: To investigate the prevalence of hearing loss after concomitant radiochemotherapy in patients enrolled in a larynx preservation protocol. DESIGN: Prospective study. SETTING: Consecutive patients treated in a tertiary cancer center hospital between 2001 and 2002. PATIENTS: Eligible subjects included patients prospectively enrolled in an organ preservation protocol based on concomitant radiotherapy and chemotherapy (cisplatin and paclitaxel). MAIN OUTCOME MEASURES: Descriptive analysis of the results of audiologic evaluations, including pure-tone audiometry and immitance audiometry, which were performed prior to and 8 months after treatment. Change in hearing sensitivity was computed relative to baseline measures. Criteria to indicate hearing decrease after the treatment were defined as either a 20-dB decrease at any single test frequency or a 10-dB decrease at any 2 adjacent test frequencies. RESULTS: A total of 11 patients were analyzed. Four patients (36%) had hearing loss after the treatment. CONCLUSION: Our results suggest that the prevalence of hearing loss after radiochemotherapy in larynx preservation protocols is high (36%); however, it was usually mild and asymptomatic. PMID- 15545580 TI - The economic cost of squamous cell cancer of the head and neck: findings from linked SEER-Medicare data. AB - OBJECTIVE: To evaluate the excess mortality, resource use, and costs associated with squamous cell carcinoma of the head and neck (SCCHN) among elderly Medicare beneficiaries. DESIGN: Retrospective cohort analysis using data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute and Medicare claims. SUBJECTS: Study cohorts included patients aged 65 years and older who were newly diagnosed as having SCCHN in a SEER registry between 1991 and 1993 (N = 4536) and controls matched 1:1 by age and sex. Patients were followed up for 5 years or until death, whichever occurred first. RESULTS: Initial treatment was primarily surgery and/or radiation among patients with early-stage SCCHN, with only modest use of chemotherapy. Patients with SCCHN had significantly (P<.001) higher 5-year mortality (64% vs 25%) and health care costs than controls. Average Medicare payments (1998 US dollars) among patients with SCCHN were $25 542 higher than those of matched comparison patients (P<.001), with monthly payments 3 times as high ($1428 vs $446). Patients diagnosed as having advanced SCCHN had shorter survival times (5-year mortality, 85%, 75%, 47%, and 35% among patients diagnosed as having distant, regional, local, and in situ cancer, respectively) and higher costs (average total Medicare payments, $53 741, $58 387, $42 698, and $37 434, respectively). CONCLUSION: These results suggest that the health economic burden of SCCHN is substantial, with costs that are comparable with or higher than those of other solid tumors. PMID- 15545581 TI - Patient, caregiver, and surgeon perceptions of quality of life following anterior skull base surgery. AB - OBJECTIVE: To estimate the agreement between surgeons' and lay caregivers' quality-of-life (QOL) perceptions of patients undergoing major skull base surgery. DESIGN: Cohort survey of patients who had undergone anterior skull base tumor excision. SETTING: University-affiliated medical center. PARTICIPANTS: Thirty-five patients and their lay caregivers participated in the study. MAIN OUTCOME MEASURES: A triple survey was performed: each patient and his or her caregiver were asked to answer 35 questions related to 6 distinct QOL domains: role of performance, physical function, vitality, pain, specific symptoms, and effect on emotions. The composite health-related QOL of the patients was also rated on an ordinal scale by 3 surgeons who participated in the operation and follow-up. RESULTS: An overall significant agreement was found between patients' and caregivers' scores at the group level (mean scores of each domain) and individual level (patient-caregiver pairs) (r = 0.76, P<.001). There was a minor correlation in the effect on emotions domain and no correlation in the pain domain. We found no correlation between the surgeons' and patients' ratings. The operating surgeons tended to overate their patients' QOL. CONCLUSIONS: The study results show that the surgeon's perception of his or her patient's QOL is not sufficiently accurate to correctly estimate patients' QOL status. These judgments should come from the patient or from the caregiver, whose perception can be used in clinical trials as a proxy for estimation of a patient's QOL. PMID- 15545582 TI - Evolutions in the management of congenital intranasal skull base defects. AB - BACKGROUND: Congenital skull base defects have traditionally been treated via an intracranial approach. Recent advances in endoscopic management have made minimally invasive extracranial approaches feasible, with less morbidity. OBJECTIVE: To determine the success of endoscopic treatment of congenital cerebrospinal fluid leaks and encephaloceles. MAIN OUTCOME MEASURES: Retrospective review of congenital cerebrospinal fluid leaks and encephaloceles treated from January 1, 1992, to December 31, 2003. Data collected include demographic characteristics, presenting signs/symptoms, site of the skull base defect, surgical approach, repair technique, and clinical follow-up. RESULTS: Eight patients were treated via the endoscopic approach for congenital cerebrospinal fluid leaks and encephaloceles. The average age at presentation was 6 years (range, birth to 18 years). Three patients presented with meningitis (average age, 6 years), 4 had cerebrospinal fluid rhinorrhea, and 3 developed a nasal obstruction. Five defects originated at the foramen cecum, and 3 others involved the ethmoid roof/cribriform plate only. Our endoscopic approaches were successful on the first attempt, with a mean follow-up of 19 months. One patient experienced nasal stenosis postoperatively. CONCLUSIONS: Continuing progress in the surgical management of congenital skull base defects demonstrates that endoscopic repair is a successful alternative to traditional craniotomy approaches, with less morbidity. This technique requires meticulous preparation and precise grafting of the defect to avoid collateral damage to surrounding structures. While reduction in the risk of meningitis, intracranial complications, and facial growth abnormalities and alleviation of nasal obstruction necessitate the timely repair of these skull base defects, special considerations are discussed regarding the optimal timing of surgical intervention, operative working space, and exposure in a smaller nasal cavity. PMID- 15545583 TI - A comparison of ocular protective measures during carbon dioxide laser laryngoscopy. AB - OBJECTIVE: To determine the efficacy of various eye protection measures during carbon dioxide laser laryngoscopy. DESIGN: A standard medical mannequin was equipped with indicator paper over the eyes and subjected to multiple passes of direct carbon dioxide laser beam contact at 400-mm focal length with powers ranging from 2.5 to 4.0 W during simulated laser microlaryngoscopy. Several different eye protection materials, including silk tape, paper tape, cloth tape, occlusive dressing, and eye pads, were used to cover the eyes and tested for their degree of protection against the laser beam. Thermal injuries were quantified and compared among these protective materials. SETTING: Academic medical center. MAIN OUTCOME MEASURE: Degree of eye protection against the laser beam injury on a scale of 0 to 4. RESULTS: The carbon dioxide laser beam at both 2.5 and 4.0 W produced considerable thermal damage to the indicator paper in the absence of any protective barrier. Among the adhesive protective barriers, moistened cloth tape was the most effective adhesive material against laser beam induced thermal damage (P<.001). Silk tape and paper tape offered poor protection. Moist eye pads, although not adhesive and therefore unable to maintain eye closure, were very effective barriers against the laser beam. Dry paper tape and dry eye pads were imminently flammable. CONCLUSION: A combination of moistened cloth tape to maintain eye closure coupled with placement of well- moistened cotton-based eye pads over the tape provides excellent eye protection during carbon dioxide laser laryngoscopy. PMID- 15545584 TI - Lack of association between esophageal biopsy, bronchoalveolar lavage, and endoscopy findings in hoarse children. AB - OBJECTIVE: To determine the prevalence of esophagitis (based on esophageal biopsy results) and aspiration (based on bronchoalveolar lavage [BAL]) in children with hoarseness. DESIGN: Retrospective medical chart review spanning 24 months of 127 consecutive children (mean age, 6.9 years; range, 1.8-17 years) who presented with hoarseness to 2 attending otolaryngologists. SETTING: Tertiary care children's hospital.Intervention All subjects underwent direct laryngoscopy, rigid bronchoscopy with BAL, and rigid or flexible esophagoscopy with biopsy. MAIN OUTCOME MEASURES: The BAL result was considered positive if the number of lipid-laden macrophages was "moderate" or "large," and the esophageal biopsy result was considered positive if any 2 of the following 3 histologic criteria were present: basal cell hyperplasia, increased papillary height, and intraepithelial inflammatory infiltrate. Comparisons between subjective endoscopic findings and objective test results were made using the t test and contingency table analysis, where appropriate. RESULTS: Of the 127 children, 104 (82%) had vocal nodules; 53 (43%) had endoscopically visualized laryngitis; 36 (28%) had tracheobronchial inflammatory changes; 60 (47%) had abnormal esophagoscopy findings; 47 (37%) had a positive BAL result; and 38 (30%) had a positive esophageal biopsy result. There was no significant correlation between BAL and esophageal biopsy results (P = .11). The odds of having positive BAL or esophageal biopsy results were unaffected by the presence of vocal nodules; endoscopically visualized inflammation of the larynx, trachea, or esophagus; or symptoms or previous clinical history of gastroesophageal reflux disease. CONCLUSION: Positive esophageal biopsy and BAL results are prevalent among children with hoarseness, regardless of subjective upper aerodigestive tract endoscopic findings. PMID- 15545585 TI - Tracheal anomalies in Pfeiffer syndrome. AB - OBJECTIVE: To determine the types and frequency of airway anomalies in patients with Pfeiffer syndrome. DESIGN: Retrospective case series. SETTING: Academic tertiary care pediatric hospital. PARTICIPANTS: Eleven patients with Pfeiffer syndrome, 6 of whom were severely affected, were identified. All were included in the study. MAIN OUTCOME MEASURES: Presence of tracheal anomalies, need for tracheotomy, and length of life. RESULTS: The 6 severely affected patients had mutations in genes that code for fibroblast growth factor receptor 2 (S351C [3 patients]; C342S [2 patients]; and W290C [1 patient]). Five of these patients were diagnosed during bronchoscopy or tracheotomy as having a congenital tracheal cartilaginous sleeve. In 1 patient, supportive care was withdrawn at 2 weeks of life, and the patient died. The remaining 5 patients required tracheotomy because of severe upper airway obstruction. Three of these patients died (at ages 9 months and 7 and 15 years). Two are still alive at ages 23 and 18 months. CONCLUSIONS: Patients with Pfeiffer syndrome manifest significant airway pathologic conditions. Upper airway obstruction is related to midface hypoplasia and secondary nasal obstruction. Tracheal anomalies have been infrequently reported. PMID- 15545586 TI - Randomized, controlled, multisite study of intracapsular tonsillectomy using low temperature plasma excision. AB - OBJECTIVE: To determine the efficacy of intracapsular tonsillectomy using low temperature plasma excision for improving the quality of the postoperative experience and for treating obstructive symptoms through 12 months postoperatively. DESIGN: Prospective, randomized, controlled, single-blind study. SETTING: Multiple private or institutional otolaryngology clinics. PATIENTS: Fifty-five children (aged 3-12 years) with obstructive tonsillar hypertrophy. INTERVENTION: Patients were randomly assigned and blinded to undergo either intracapsular tonsillectomy using low-temperature plasma excision (n = 27) or total tonsillectomy using conventional electrosurgery (n = 28). MAIN OUTCOME MEASURES: Operative data, 14-day recovery variables, and obstructive symptoms were prospectively collected through 12 months. RESULTS: During the first 14 days, significantly fewer children in the intracapsular group reported nausea (P = .01) or lost weight (P = .003). The intracapsular group had a significantly faster resolution of pain (P = .01), had an earlier return to a normal diet (P = .004), ceased taking pain medication sooner (P = .002), and returned to normal activity sooner (P = .04). Postoperatively, the intracapsular group had more residual tonsil tissue than the total tonsillectomy group (P = .002 for the 3- and 12-month visits). However, the incidence of recurring obstructive symptoms, pharyngitis, and antibiotic use was similar in both treatment groups during the 12 months. CONCLUSIONS: Postoperative morbidity normally associated with traditional (total) tonsillectomy was significantly reduced after intracapsular tonsillectomy using low-temperature plasma excision. The residual tonsillar tissue associated with this technique was of no clinical consequence. PMID- 15545587 TI - Radiation therapy does not impact local complication rates after free flap reconstruction for head and neck cancer. AB - OBJECTIVE: To determine whether external beam radiation therapy (XRT), administered either before or after surgery, increases the rate and/or severity of local postoperative complications in patients with head and neck cancer who undergo microvascular free flap reconstruction. DESIGN: Retrospective cohort study. SETTING: University of Washington Medical Center, Seattle, a tertiary care hospital. PATIENTS: A total of 100 consecutive patients underwent fibular free flap reconstruction of the mandible. The study cohort was divided according to radiation treatment status: (1) no XRT (28 patients), (2) preoperative XRT (37 patients), and (3) postoperative XRT (35 patients). The median follow-up after surgery was 11 months (range, 1-89 months). MAIN OUTCOME MEASURES: Rate and severity of local postoperative complications. RESULTS: Fifty-four patients (54%) had at least 1 postoperative complication. There were no differences among the 3 XRT subgroups in the overall proportion of patients with complications of any severity (15 [54%] of 28 patients in the no XRT group, 24 [65%] of 37 patients in the preoperative XRT group, and 16 [46%] of 35 patients in the postoperative XRT group; P = .26, chi(2) analysis). There were also no differences seen when mild and severe complication rates were specifically examined (P = .58 and P = .10, respectively). No case of complete flap loss was observed. We noted no significant correlations between the rate of postoperative complications and the following covariates: total radiation dose, size of radiation field, disease stage, exposure to chemotherapy, presence of serious medical comorbidities, patient age, or history of tobacco use. CONCLUSIONS: Our experience suggests that XRT can be safely administered before or after surgery to patients undergoing head and neck free flap reconstruction at an experienced surgical referral center. Postoperative complication rates were not significantly affected by administration, timing, dose, or extent of XRT. PMID- 15545588 TI - The effect on snoring of structural nasal valve dilatation with a butterfly graft. AB - OBJECTIVE: To evaluate the effect on snoring of structural nasal valve dilatation with butterfly spreader grafts in patients with nasal valve insufficiency. DESIGN: Retrospective medical chart review and telephone follow-up; mean +/- SD follow-up time, 20.7 +/- 11.34 months (range, 3-48 months). SETTINGS: Tertiary care referral center. SUBJECTS: A total of 37 snoring patients with nasal valve insufficiency who underwent nasal valve dilatation with a butterfly spreader graft. INTERVENTIONS: The conchal cartilage butterfly graft technique was performed during rhinoplasty through either an external or endonasal approach. MAIN OUTCOME MEASURE: To establish through a retrospective review that butterfly graft conchal cartilage nasal reconstruction is effective in reducing snoring. RESULTS: After surgery, 30 patients (81%) had significant improvement in breathing, 5 (14%) had slight improvement, and 2 (5%) had no benefit in breathing. Snoring stopped completely in 11 (30%) of the patients after surgery. The improvement in snoring was significant in 13 patients (35%) and slight in 3 (8%). Twenty-six patients (70%) reported tiredness and grogginess on awakening before the surgery. Surgery significantly improved patients' tiredness and grogginess on awakening in 15 cases (58%), slightly improved them in 5 (19%), and did not change the patients' tiredness and grogginess in 6 cases (23%). CONCLUSION: The conchal cartilage butterfly graft yields successful results not only in breathing but also in snoring symptoms in patients with nasal valve insufficiency. PMID- 15545589 TI - Intralesional cidofovir for pediatric recurrent respiratory papillomatosis. AB - OBJECTIVE: To compare disease response among children with recurrent respiratory papillomatosis (RRP) who underwent combined surgical debulking and intralesional cidofovir injections vs repeated surgical debulking only. DESIGN: Retrospective medical record review; follow-up range: 16 to 56 months. SETTING: Tertiary care children's hospital. PATIENTS: Seven children with RRP, including 4 subjects treated with cidofovir and 3 controls matched for age and initial papilloma staging score. INTERVENTIONS: Subjects treated with cidofovir underwent combined surgical debulking and intralesional cidofovir injection every 2 months until disease remission. Control subjects underwent repeated surgical debulking at individually determined intervals. MAIN OUTCOME MEASURES: Intraoperative endoscopic photographs were retrospectively assigned papilloma staging scores. Cidofovir and control group comparisons were made using nonparametric 2-sample Wilcoxon rank-sum (Mann-Whitney) testing, and comparisons of initial and final papilloma staging scores were made using nonparametric matched-pair Wilcoxon signed-rank testing. RESULTS: The final cidofovir group scores were significantly lower than the control group scores (P < .05). Within-group differences between initial and final scores were not significant (cidofovir group, P = .07; control group, P = .29). CONCLUSIONS: Four children with RRP were safely and successfully treated with intralesional cidofovir injection. Consideration should be given to using cidofovir more widely for treatment of pediatric RRP. Larger numbers in the cidofovir and control groups are needed in future studies to determine the true impact of cidofovir on management of this disease. PMID- 15545590 TI - Subcutaneous emphysema and pneumomediastinum complicating tonsillectomy. AB - BACKGROUND: Subcutaneous emphysema and pneumomediastinum are very rare complications of tonsillectomy. OBJECTIVES: To report on a case study and a literature review of these conditions; to discuss the management and controversial causes. DESIGN: Case study and literature review. METHODS: A case is presented of subcutaneous emphysema and pneumomediastinum complicating tonsillectomy. All published reports of these complications dating back to 1910 are reviewed. RESULTS: An additional 30 cases of subcutaneous emphysema and 9 of pneumomediastinum have been reported following tonsillectomy. This group of 32 patients with emphysematous complications included 17 male patients, 14 female, and a child of unknown sex. The mean age was 18.6 years (range, 2-65 years). All tonsillectomies were performed with a cold technique except our case. Subcutaneous emphysema was first noted intraoperatively in 8 patients (25%) a mean of 3.5 hours (range, 10 minutes to 14 hours) after the procedure in 18 patients (56%) and not until the next morning in 6 patients (19%). Most cases resolved without specific treatment or significant sequelae. Some cases were life threatening and required urgent intervention. Others were misdiagnosed, which resulted in unnecessary surgery. CONCLUSION: Subcutaneous emphysema and pneumomediastinum are rare occurrences following tonsillectomy that should alert one to the possibility of more serious complications. PMID- 15545591 TI - What are the optimal techniques for skin rejuvenation? PMID- 15545592 TI - Defining clinical responses in cosmetic laser surgery. PMID- 15545593 TI - Proven modalities for facial skin resurfacing? PMID- 15545594 TI - Pathology quiz case 1. Angiolymphoid hyperplasia with eosinophilia. PMID- 15545595 TI - Pathology quiz case 2. First branchial cleft cyst. PMID- 15545596 TI - Radiology quiz case 1. Rhinolith. PMID- 15545597 TI - Radiology quiz case 2. Hypervascular vagal schwannoma mimicking paraganglioma. PMID- 15545598 TI - Molecular evolution of dinoflagellate luciferases, enzymes with three catalytic domains in a single polypeptide. AB - Enzymes with multiple catalytic sites are rare, and their evolutionary significance remains to be established. This study of luciferases from seven dinoflagellate species examines the previously undescribed evolution of such proteins. All these enzymes have the same unique structure: three homologous domains, each with catalytic activity, preceded by an N-terminal region of unknown function. Both pairwise comparison and phylogenetic inference indicate that the similarity of the corresponding individual domains between species is greater than that between the three different domains of each polypeptide. Trees constructed from each of the three individual domains are congruent with the tree of the full-length coding sequence. Luciferase and ribosomal DNA trees both indicate that the Lingulodinium polyedrum luciferase diverged early from the other six. In all species, the amino acid sequence in the central regions of the three domains is strongly conserved, suggesting it as the catalytic site. Synonymous substitution rates also are greatly reduced in the central regions of two species but not in the other five. This lineage-specific difference in synonymous substitution rates in the central region of the domains correlates inversely with the content of GC3, which can be accounted for by the biased usage toward C-ending codons at the degenerate sites. RNA modeling of the central region of the L. polyedrum luciferase domain suggests a function of the constrained synonymous substitutions in the circadian-controlled protein synthesis. PMID- 15545599 TI - TNF receptor-associated factor (TRAF) domain and Bcl-2 cooperate to induce small B cell lymphoma/chronic lymphocytic leukemia in transgenic mice. AB - Transgenic mice overexpressing in B lymphocytes either Bcl-2 or a TNF receptor associated factor (TRAF)2 mutant lacking the N-terminal RING and zinc finger domains located at the N terminus of the molecule (TRAF2DN), which mimics TRAF1, developed lymphadenopathy and splenomegaly due to polyclonal B cell expansion. Remarkably, TRAF2DN/Bcl-2 double-transgenic mice contained B cell populations similar to those observed in TRAF2DN mice. However, over time, they developed severe splenomegaly and lymphadenopathy, and most animals also developed leukemia, pleural effusion, and, in some cases, ascites associated with monoclonal and oligoclonal B cell neoplasms. The life span of TRAF2DN/Bcl-2 mice was markedly reduced compared with Bcl-2 and TRAF2DN single-transgenics or wild type littermates. The expanded B cell population of TRAF2DN/Bcl-2 double transgenic mice was primarily comprised of small/medium-size noncycling B220(M)/IgM(H)/IgD(L)/CD21(L)/CD23(NULL)/CD11b(+)/CD5+ cells that were Bcl-6 negative, consistent with a B-1 phenotype. The cells also expressed high levels of CD54 and other adhesion molecules. In vitro, these B cells showed comparable proliferation rates to those of wild-type counterparts but exhibited markedly increased survival and were resistant to apoptosis induced by chemotherapeutic agents and glucocorticoids. Histopathologic features were consistent with mouse small B cell lymphoma progressing to leukemia with many similarities to human chronic lymphocytic leukemia. Given that many human chronic lymphocytic leukemias overexpress TRAF1 and Bcl-2, our findings suggest that cooperation between Bcl-2 and TRAF pathways contributes to the development of this type of leukemia. PMID- 15545600 TI - The role of the glycocalyx in reorganization of the actin cytoskeleton under fluid shear stress: a "bumper-car" model. AB - We propose a conceptual model for the cytoskeletal organization of endothelial cells (ECs) based on a major dichotomy in structure and function at basal and apical aspects of the cells. Intracellular distributions of filamentous actin (F actin), vinculin, paxillin, ZO-1, and Cx43 were analyzed from confocal micrographs of rat fat-pad ECs after 5 h of shear stress. With intact glycocalyx, there was severe disruption of the dense peripheral actin bands (DPABs) and migration of vinculin to cell borders under a uniform shear stress (10.5 dyne/cm2; 1 dyne = 10 microN). This behavior was augmented in corner flow regions of the flow chamber where high shear stress gradients were present. In striking contrast, no such reorganization was observed if the glycocalyx was compromised. These results are explained in terms of a "bumper-car" model, in which the actin cortical web and DPAB are only loosely connected to basal attachment sites, allowing for two distinct cellular signaling pathways in response to fluid shear stress, one transmitted by glycocalyx core proteins as a torque that acts on the actin cortical web (ACW) and DPAB, and the other emanating from focal adhesions and stress fibers at the basal and apical membranes of the cell. PMID- 15545601 TI - Activation of RasGRP3 by phosphorylation of Thr-133 is required for B cell receptor-mediated Ras activation. AB - The Ras signaling pathway plays a critical role in B lymphocyte development and activation, but its activation mechanism has not been well understood. At least one mode of Ras regulation in B cells involves a Ras-guanyl nucleotide exchange factor, RasGRP3. We demonstrate here that RasGRP3 undergoes phosphorylation at Thr-133 upon B cell receptor cross-linking, thereby resulting in its activation. Deletion of phospholipase C-gamma2 or pharmacological interference with conventional PKCs resulted in marked reduction in both Thr-133 phosphorylation and Ras activation. Moreover, mutation of Thr-133 in RasGRP3 alone severely impaired its ability to activate Ras in B cell receptor signaling. Hence, our data suggest that PKC, after being activated by diacylglycerol, phosphorylates RasGRP3, thereby contributing to its full activation. PMID- 15545602 TI - A genetic screen to isolate type III effectors translocated into pepper cells during Xanthomonas infection. AB - The bacterial pathogen Xanthomonas campestris pv. vesicatoria (Xcv) uses a type III secretion system (TTSS) to translocate effector proteins into host plant cells. The TTSS is required for Xcv colonization, yet the identity of many proteins translocated through this apparatus is not known. We used a genetic screen to functionally identify Xcv TTSS effectors. A transposon 5 (Tn5)-based transposon construct including the coding sequence for the Xcv AvrBs2 effector devoid of its TTSS signal was randomly inserted into the Xcv genome. Insertion of the avrBs2 reporter gene into Xcv genes coding for proteins containing a functional TTSS signal peptide resulted in the creation of chimeric TTSS effector::AvrBs2 fusion proteins. Xcv strains containing these fusions translocated the AvrBs2 reporter in a TTSS-dependent manner into resistant BS2 pepper cells during infection, activating the avrBs2-dependent hypersensitive response (HR). We isolated seven chimeric fusion proteins and designated the identified TTSS effectors as Xanthomonas outer proteins (Xops). Translocation of each Xop was confirmed by using the calmodulin-dependent adenylate cydase reporter assay. Three xop genes are Xanthomonas spp.-specific, whereas homologs for the rest are found in other phytopathogenic bacteria. XopF1 and XopF2 define an effector gene family in Xcv. XopN contains a eukaryotic protein fold repeat and is required for full Xcv pathogenicity in pepper and tomato. The translocated effectors identified in this work expand our knowledge of the diversity of proteins that Xcv uses to manipulate its hosts. PMID- 15545603 TI - Structure and function of the phenazine biosynthetic protein PhzF from Pseudomonas fluorescens. AB - Phenazines produced by Pseudomonas and Streptomyces spp. are heterocyclic nitrogen-containing metabolites with antibiotic, antitumor, and antiparasitic activity. The antibiotic properties of pyocyanin, produced by Pseudomonas aeruginosa, were recognized in the 1890s, although this blue phenazine is now known to be a virulence factor in human disease. Despite their biological significance, the biosynthesis of phenazines is not fully understood. Here we present structural and functional studies of PhzF, an enzyme essential for phenazine synthesis in Pseudomonas spp. PhzF shares topology with diaminopimelate epimerase DapF but lacks the same catalytic residues. The structure of PhzF in complex with its substrate, trans-2,3-dihydro-3-hydroxyanthranilic acid, suggests that it is an isomerase using the conserved glutamate E45 to abstract a proton from C3 of the substrate. The proton is returned to C1 of the substrate after rearrangement of the double-bond system, yielding an enol that converts to the corresponding ketone. PhzF is a dimer that may be bifunctional, providing a shielded cavity for ketone dimerization via double Schiff-base formation to produce the phenazine scaffold. Our proposed mechanism is supported by mass and NMR spectroscopy. The results are discussed in the context of related structures and protein sequences of unknown biochemical function. PMID- 15545604 TI - Integrative analysis of genome-scale data by using pseudoinverse projection predicts novel correlation between DNA replication and RNA transcription. AB - We describe an integrative data-driven mathematical framework that formulates any number of genome-scale molecular biological data sets in terms of one chosen set of data samples, or of profiles extracted mathematically from data samples, designated the "basis" set. By using pseudoinverse projection, the molecular biological profiles of the data samples are least-squares-approximated as superpositions of the basis profiles. Reconstruction of the data in the basis simulates experimental observation of only the cellular states manifest in the data that correspond to those of the basis. Classification of the data samples according to their reconstruction in the basis, rather than their overall measured profiles, maps the cellular states of the data onto those of the basis and gives a global picture of the correlations and possibly also causal coordination of these two sets of states. We illustrate this framework with an integration of yeast genome-scale proteins' DNA-binding data with cell cycle mRNA expression time course data. Novel correlation between DNA replication initiation and RNA transcription during the yeast cell cycle, which might be due to a previously unknown mechanism of regulation, is predicted. PMID- 15545605 TI - Fluorescent indicators of cAMP and Epac activation reveal differential dynamics of cAMP signaling within discrete subcellular compartments. AB - Second messenger cAMP regulates many cellular functions through its effectors, such as cAMP-dependent protein kinase (PKA) and Epac (exchange proteins directly activated by cAMP). Spatial and temporal control of cAMP signaling is crucial to differential regulation of cellular targets involved in various signaling cascades. To investigate the compartmentalized cAMP signaling, we constructed fluorescent indicators that report intracellular cAMP dynamics and Epac activation by sandwiching the full-length Epac1 between cyan and yellow mutants of GFP. Elevations of cAMP decreased FRET and increased the ratio of cyan-to yellow emissions by 10-30% in living mammalian cells. This response can be reversed by removing cAMP-elevating agents and abolished by mutating the critical residue responsible for cAMP binding. Targeting of the reporter to the plasma membrane, where cAMP is produced in response to the activation of beta-adrenergic receptor, revealed a faster cAMP response at the membrane than in the cytoplasm and mitochondria. Simultaneous imaging with targeted cAMP indicator and PKA activity reporter allowed the detection of a much delayed PKA response in the nucleus after the rapid accumulation of cAMP at the plasma membrane of the same cell, despite the immediate presence of a pool of cAMP in the nucleus. Thus, cAMP dynamics and the activation of its effectors are precisely controlled spatiotemporally in vivo. PMID- 15545606 TI - Climate change impacts are sensitive to the concentration stabilization path. AB - Analysis of policies to achieve the long-term objective of the United Nations Framework Convention on Climate Change, stabilizing concentrations of greenhouse gases at levels that avoid "dangerous" climate changes, must discriminate among the infinite number of emission and concentration trajectories that yield the same final concentration. Considerable attention has been devoted to path dependent mitigation costs, generally for CO2 alone, but not to the differential climate change impacts implied by alternative trajectories. Here, we derive pathways leading to stabilization of equivalent CO2 concentration (including radiative forcing effects of all significant trace gases and aerosols) with a range of transient behavior before stabilization, including temporary overshoot of the final value. We compare resulting climate changes to the sensitivity of representative geophysical and ecological systems. Based on the limited available information, some physical and ecological systems appear to be quite sensitive to the details of the approach to stabilization. The likelihood of occurrence of impacts that might be considered dangerous increases under trajectories that delay emissions reduction or overshoot the final concentration. PMID- 15545607 TI - Mitochondrial biogenesis by NO yields functionally active mitochondria in mammals. AB - We recently found that long-term exposure to nitric oxide (NO) triggers mitochondrial biogenesis in mammalian cells and tissues by activation of guanylate cyclase and generation of cGMP. Here, we report that the NO/cGMP dependent mitochondrial biogenesis is associated with enhanced coupled respiration and content of ATP in U937, L6, and PC12 cells. The observed increase in ATP content depended entirely on oxidative phosphorylation, because ATP formation by glycolysis was unchanged. Brain, kidney, liver, heart, and gastrocnemius muscle from endothelial NO synthase null mutant mice displayed markedly reduced mitochondrial content associated with significantly lower oxygen consumption and ATP content. In these tissues, ultrastructural analyses revealed significantly smaller mitochondria. Furthermore, a significant reduction in the number of mitochondria was observed in the subsarcolemmal region of the gastrocnemius muscle. We conclude that NO/cGMP stimulates mitochondrial biogenesis, both in vitro and in vivo, and that this stimulation is associated with increased mitochondrial function, resulting in enhanced formation of ATP. PMID- 15545608 TI - Medicinal chemistry applied to a synthetic protein: development of highly potent HIV entry inhibitors. AB - We have used total chemical synthesis to perform high-resolution dissection of the pharmacophore of a potent anti-HIV protein, the aminooxypentane oxime of [glyoxylyl1]RANTES(2-68), known as AOP-RANTES, of which we designed and made 37 analogs. All involved incorporation of one or more rationally chosen nonnatural noncoded structures, for which we found a clear comparative advantage over coded ones. We investigated structure-activity relationships in the pharmacophore by screening the analogs for their ability to block the HIV entry process and produced a derivative, PSC-RANTES [N-nonanoyl, des-Ser1[L-thioproline2, L cyclohexylglycine3]-RANTES(2-68)], which is 50 times more potent than AOP-RANTES. This promising group of compounds might be optimized yet further as potential prophylactic and therapeutic anti-HIV agents. The remarkable potency of our RANTES analogs probably involves the unusual mechanism of intracellular sequestration of CC-chemokine receptor 5 (CCR5), and it has been suggested that this arises from enhanced affinity for the receptor. We found that inhibitory potency and capacity to induce CCR5 down-modulation do appear to be correlated, but that unexpectedly, inhibitory potency and affinity for CCR5 do not. We believe this study represents the proof of principle for the use of a medicinal chemistry approach, above all one showing the advantage of noncoded structures, to the optimization of the pharmacological properties of a protein. Medicinal chemistry of small molecules is the foundation of modern pharmaceutical practice, and we believe we have shown that techniques have now reached the point at which the approach could also be applied to the many macromolecular drugs now in common use. PMID- 15545609 TI - Gestational exposure of Ahr and Arnt hypomorphs to dioxin rescues vascular development. AB - The aryl hydrocarbon receptor (AHR) is commonly known for its role in the adaptive metabolism of xenobiotics and in the toxic events that follow exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin). Previously, we have demonstrated that the AHR and its heterodimeric partner, the AHR nuclear translocator (ARNT), play a role in the developmental closure of a hepatic vascular shunt known as the ductus venosus (DV). To investigate the mechanism of DV closure, we generated hypomorphic alleles of the Ahr and Arnt loci. Using these models, we then asked whether this vascular defect could be rescued by receptor activation during late development. By manipulating gestational exposure, the patent DV in AHR or ARNT hypomorphs could be efficiently closed by dioxin exposure as early as embryonic day 12.5 and as late as embryonic day 18.5. These findings define the temporal regulation of receptor activation during normal ontogeny and provide evidence to support the idea that receptor activation and AHR-ARNT heterodimerization are essential for normal vascular development. Taken in the broader context, these data demonstrate that similar AHR signaling steps govern all major aspects of AHR biology. PMID- 15545610 TI - Long-range compaction and flexibility of interphase chromatin in budding yeast analyzed by high-resolution imaging techniques. AB - Little is known about how chromatin folds in its native state. Using optimized in situ hybridization and live imaging techniques have determined compaction ratios and fiber flexibility for interphase chromatin in budding yeast. Unlike previous studies, ours examines nonrepetitive chromatin at intervals short enough to be meaningful for yeast chromosomes and functional domains in higher eukaryotes. We reconcile high-resolution fluorescence in situ hybridization data from intervals of 14-100 kb along single chromatids with measurements of whole chromosome arms (122-623 kb in length), monitored in intact cells through the targeted binding of bacterial repressors fused to GFP derivatives. The results are interpreted with a flexible polymer model and suggest that interphase chromatin exists in a compact higher-order conformation with a persistence length of 170-220 nm and a mass density of approximately 110-150 bp/nm. These values are equivalent to 7-10 nucleosomes per 11-nm turn within a 30-nm-like fiber structure. Comparison of long and short chromatid arm measurements demonstrates that chromatin fiber extension is also influenced by nuclear geometry. The observation of this surprisingly compact chromatin structure for transcriptionally competent chromatin in living yeast cells suggests that the passage of RNA polymerase II requires a very transient unfolding of higher-order chromatin structure. PMID- 15545611 TI - Identification and functional characterization of a sex pheromone receptor in the silkmoth Bombyx mori. AB - Sex pheromones released by female moths are detected with high specificity and sensitivity in the olfactory sensilla of antennae of conspecific males. Bombykol in the silkmoth Bombyx mori was the first sex pheromone to be identified. Here we identify a male-specific G protein-coupled olfactory receptor gene, B. mori olfactory receptor 1 (BmOR-1), that appears to encode a bombykol receptor. The BmOR-1 gene is located on the Z sex chromosome, has an eight-exon/seven-intron structure, and exhibits male-specific expression in the pheromone receptor neurons of male moth antenna during late pupal and adult stages. Bombykol stimulation of Xenopus laevis oocytes expressing BmOR-1 and BmGalphaq elicited robust dose-dependent inward currents on two-electrode voltage clamp recordings, demonstrating that the binding of bombykol to BmOR-1 leads to the activation of a BmGalphaq-mediated signaling cascade. Antennae of female moths infected with BmOR 1-recombinant baculovirus showed electrophysiological responses to bombykol but not to bombykal. These results provide evidence that BmOR-1 is a G protein coupled sex pheromone receptor that recognizes bombykol. PMID- 15545612 TI - On the evolutionary history of Ephedra: Cretaceous fossils and extant molecules. AB - Gnetales comprise three unusual genera of seed plants, Ephedra, Gnetum, and Welwitschia. Their extraordinary morphological diversity suggests that they are survivors of an ancient, more diverse group. Gnetalean antiquity is also supported by fossil data. Dispersed "ephedroid" (polyplicate) pollen first appeared in the Permian >250 million years ago (Myr), and a few megafossils document the presence of gnetalean features in the early Cretaceous. The Cretaceous welwitschioid seedling Cratonia cotyledon dates the split between Gnetum and Welwitschia to before 110 Myr. Ages and character evolution of modern diversity are, however, controversial, and, based on molecular data, it has recently been suggested that Ephedra is very young, only 8-32 Myr. Here, we present data on the evolutionary history of Ephedra. Fossil seeds from Buarcos, Portugal, unequivocally link one type of Cretaceous polyplicate pollen to Ephedra and document that plants with unique characters, including the peculiar naked male gametophyte, were established already in the Early Cretaceous. Clades in our molecular phylogeny of extant species correspond to geographical regions, with African species in a basal grade/clade. The study demonstrates extremely low divergence in both molecular and morphological characters in Ephedra. Features observed in the fossils are present in all major extant clades, showing that modern species have retained unique reproductive characters for >110 million years. A recent origin of modern species of Ephedra would imply that the Cretaceous Ephedra fossils discussed here were members of widespread, now extinct sister lineage(s), and that no morphological innovations characterized the second diversification. PMID- 15545613 TI - TATA box binding protein induces structure in the recombinant glucocorticoid receptor AF1 domain. AB - A number of transcription factor proteins contain domains that are fully or partially unstructured. The means by which such proteins acquire naturally folded conformations are not well understood. When they encounter their proper binding partner(s), several of these proteins adopt a folded conformation through an induced-fit mechanism. The glucocorticoid receptor (GR) is a ligand-activated transcription factor. Expressed independently as a recombinant peptide, the N terminal transactivation domain (AF1) of the GR shows little structure and appears to exist as a collection of random coil configurations. The GR AF1 is known to interact with other transcription factors, including a critical component of the general transcription machinery proteins, the TATA box binding protein (TBP). We tested whether this interaction can lead to acquisition of structure in the GR AF1. Our results show that recombinant GR AF1 acquires a significant amount of helical content when it interacts with TBP. These structural changes were monitored by Fourier transform infrared and NMR spectroscopies, and by proteolytic digestions. Our results support a model in which TBP binding interaction with the GR AF1 induces significantly greater helical structure in the AF1 domain. This increased helical content in the GR AF1 appears to come mostly at the expense of random coil conformation. These results are in accordance with the hypothesis that an induced-fit mechanism gives structure to the GR AF1 when it encounters TBP. PMID- 15545614 TI - A prolactin family paralog regulates reproductive adaptations to a physiological stressor. AB - Successful species develop strategies to optimize their reproductive performance. This optimization likely includes the evolution of genes that specifically permit reproduction in physiologically challenging conditions. The prolactin (PRL) family gene cluster is one of 25 mouse-specific gene clusters, the majority of which are associated with reproduction. A prevailing theme characterizing the PRL family is its connection with pregnancy and mechanisms controlling viviparity. PRL-like protein A (PLP-A) is one of 26 genes located within the PRL family locus. It is a nonclassical member of the PRL family (e.g., PLP-A does not use the PRL receptor) produced by trophoblast cells of the chorioallantoic placenta and acts on uterine natural killer cells. In this report, the biology of PLP-A has been investigated by generating mice with a PLP-A null mutation. Under standardized animal husbandry conditions, PLP-A possesses modest effects on reproductive performance. However, this same gene is critical for reproduction when mice are exposed to a physiological stressor. Wild-type mice exposed to hypobaric hypoxia during gestation readily adapt and maintain their pregnancies, whereas PLP-A null mutant mice fail to adapt, resulting in pregnancy failure. PLP A contributes to hypoxia-induced adaptations critical to hemochorial placentation and thus nutrient flow to extraembryonic and embryonic tissues. The findings provide insights into species-specific reproductive adaptations. PMID- 15545615 TI - Kikuchi-Fujimoto disease with prolonged fever in children. AB - We reviewed 12 patients who had Kikuchi-Fujimoto disease (KFD) and presented with prolonged fever and lymphadenopathy. The clinical and laboratory aspects of the patients confirmed by excisional lymph node biopsy were analyzed. The mean age of the children was 11.0 +/- 3.0 years (range: 6-15 years). The male-to-female ratio was 1.4:1. The median duration of fever before admission and the total duration of fever was 13 days (range: 7-65 days) and 19.5 days (range: 9-75 days), respectively. One patient had supraclavicular lymphadenopathy, 10 had cervical involvement, and 1 had axillary lymphadenopathy. All of the histologic findings of the lymph node biopsies showed the characteristic findings consistent with KFD, such as paracortical necrosis with karyorrhexis and an increase in the number of phagocytic histiocytes and atypical lymphocytes. As for the laboratory findings, leukopenia (3600 +/- 900 per mm3), anemia (hemoglobin 11.4 +/- 1.2 g/dL), an elevated erythrocyte sedimentation rate (44 +/- 18 mm/hour), and a relatively low C-reactive protein level (1.3 +/- 1.1 mg/dL) were noted. Eight patients received conservative therapy with antipyretics, and 3 patients were treated with prednisolone. KFD is a rare disease yet should be considered in the differential diagnosis for older children with prolonged fever and lymphadenopathy. PMID- 15545616 TI - Growth, efficacy, and safety of feeding an iron-fortified human milk fortifier. AB - OBJECTIVE: Survival rates for preterm infants who weigh between 501 and 1500 g at birth have continued to improve over time. In response to this continuing decrease in birth weight of surviving preterm infants, Enfamil Human Milk Fortifier has recently been reformulated to meet the nutritional requirements of these smaller, more rapidly growing infants. It now provides an increased protein level of 1.1 g/58 kJ, a decreased carbohydrate level of 0.2 g/58 kJ, and a combined linoleic and alpha-linolenic fatty acid content of 157 mg/58 kJ. As these very small preterm infants have an increased requirement for dietary iron, the fortifier has been supplemented with 1.44 mg/58 kJ of iron, an amount of iron similar to that provided in a typical iron-fortified term infant formula. An iron fortified product obviates the need for administration of an iron supplement, a hyperosmolar-inducing intervention. The purpose of this prospective, double blind, randomized, controlled study was to evaluate growth, safety, and efficacy in a population of very low birth weight (VLBW) preterm infants who received human milk fortified with either the reformulated iron-fortified powdered human milk fortifier test product (HMF-T) or a powdered commercially available human milk fortifier control product (HMF-C). METHODS: Infants who weighed < or =1500 g, had a gestational age < or =33 weeks postmenstrual age, and had an enteral intake of at least 100 mL/kg per day of unfortified human milk were stratified by gender and birth weight and randomized to receive HMF-T or HMF-C product from study day 1 to study day 28, hospital discharge, or the termination of human milk feedings, whichever came first. Unless medically indicated, investigators were not to administer iron supplements from study days 1 to 14. Infants were assessed serially for growth; enteral and parenteral intake; serum chemistry and hematologic values; clinical histories, including the administration of blood transfusions; feeding tolerance; respiratory outcomes; and morbidities, including adverse events. RESULTS: Of the 181 participating infants in this study, 96 received HMF-T and 85 received HMF-C. At randomization, there were no significant differences in infant characteristics between the fortifier groups. The percentage of participants who remained in the study for 28 days was similar between fortifier groups (57% HMF-T, 46% HMF-C). For both fortifier groups, the most frequent reasons for discontinuing the study before study day 28 were unavailability of human milk and hospital discharge. Rate of weight gain was similar between the fortifier groups (17.5 +/- 0.53 g/kg per day for HMF-T and 17.3 +/- 0.59 g/kg per day for HMF-C). Mean achieved weight, length, and head circumference were comparable between groups across the 28-day study period. Total protein intake from enteral and parenteral nutrition was significantly greater for the HMF-T fortifier group; however, this difference did not result in any difference in growth between the 2 fortifier groups. An analysis of the growth and energy intake data of a subset of the intent-to-treat population who adhered more strictly to the study feeding protocol yielded results similar to those seen for the intent-to-treat population. There were no clinically significant differences in the results of laboratory studies between the groups at study days 0, 14, and 28. Anemia of prematurity was prevalent in both study groups; by study day 28, median hematocrit levels were 27.0% (interquartile range [IQR]: 24.0%-29.6%) for the HMF-T group and 26.0% (IQR: 24.0%-31.0%) for the HMF C group. Median ferritin levels were 77.0 ng/mL (IQR: 37-155 ng/ml) for HMF-T and 92.0 ng/mL (IQR: 33-110 ng/mL) for HMF-C. There were no significant differences between the study fortifier groups in regard to the receipt of medically indicated iron supplements on or before study day 14 or in the administration of blood transfusions before study day 0 or from study days 0 through 14. However, from study day 15 to study day 28, fewer HMF-T infants (n = 12) required a blood transfusion than did HMF-C infants (n = 20). Although the higher levels of iron in the HMF-T fortifier (1.44 mg vs 0.35 mg for HMF-C per 4 packets of powdered fortifier) did not prevent anemia per se, it did reduce the frequency of one of the most serious outcomes of anemia: the need for a blood transfusion. There was no statistically significant difference between fortifier groups in regard to feeding tolerance. Rates of suspected sepsis (26% HMF-T vs 31% HMF-C) and confirmed sepsis (5% HMF-T, 7% HMF-C) were low as were the rates of suspected necrotizing enterocolitis (NEC; 6% HMF-T and 5% HMF-C) and confirmed Bell's stage 2 or more NEC (1% HMF-T and 1% HMF-C). There were no statistically significant differences between the study fortifier groups in regard to the incidence of confirmed and suspected sepsis and NEC. CONCLUSION: Both human milk fortifiers studied are safe, are well tolerated, and facilitate comparable good growth; however, using the iron-fortified product may reduce the need for blood transfusions in VLBW infants. The similar low rates of suspected and confirmed NEC and sepsis seen in both fortifier groups in this study refutes the premise that the inclusion of iron in fortifiers will increase the incidence of sepsis and NEC. Indeed, the incidence for NEC and sepsis for both groups in this study was lower than is reported for VLBW infants and similar to that seen for infants who are fed human milk. PMID- 15545617 TI - Treatment of acute Kawasaki disease: aspirin's role in the febrile stage revisited. AB - OBJECTIVE: To evaluate the effect of treatment without aspirin in the acute phase of Kawasaki disease (KD) and to determine whether it is necessary to expose children to high- or medium-dose aspirin. METHODS: A total of 162 patients who fulfilled the established criteria of acute KD between 1993 and 2003 were included in this retrospective study. All patients were treated with high-dose intravenous immunoglobulin (IVIG; 2 g/kg) as a single infusion without concomitant aspirin treatment. Low-dose aspirin (3-5 mg/kg per day) was subsequently prescribed when fever subsided. Patients who had defervescence within 3 days after the completion of IVIG treatment were classified as the IVIG responsive group, and those whose fever persisted for >3 days were classified as the IVIG-nonresponsive group. The 162 patients were divided further into 2 groups: those who were treated with IVIG before illness day 5, and those who were treated after illness day 5. We compared the response rate of IVIG therapy, duration of fever, and incidence of coronary artery abnormalities (CAAs) between these groups. RESULTS: A total of 153 patients were classified into the IVIG responsive group, and 128 (83.66%) of them had defervescence within 24 hours after completion of IVIG therapy. Nine (5.56%) patients were classified into the IVIG nonresponsive group, and all received additional IVIG (2 g/kg) without aspirin. Six (66.67%) had defervescence within 3 days after additional therapy. Patients in the IVIG-nonresponsive group had a significantly higher incidence of CAAs than those in the IVIG-responsive group (25% vs 2.92%). In the group that was treated before illness day 5 (n = 16), all patients had defervescence within 3 days after IVIG therapy and 13 (81.25%) had defervescence within 24 hours. In the group that was treated after illness day 5 (n = 146), 137 (93.84%) patients had defervescence within 3 days and 115 (78.77%) had defervescence within 24 hours. One (6.67%) patient in the group that was treated before illness day 5 got a new onset of CAAs, as did 5 (3.85%) in the group that was treated after illness day 5. There was no statistically significant difference in the response rate of IVIG therapy, duration of fever, and incidence of CAAs between these 2 groups. CONCLUSION: The results of our study indicate that the treatment without aspirin in acute stage of KD had no effect on the response rate of IVIG therapy, duration of fever, or incidence of CAAs when children were treated with high-dose (2 g/kg) IVIG as a single infusion, despite treatment before or after day 5 of illness. We conclude that it seems unnecessary to expose children to high- or medium-dose aspirin therapy in acute KD when the available data show no appreciable benefit in preventing the failure of IVIG therapy, formation of CAAs, or shortening the duration of fever. PMID- 15545618 TI - Echocardiographic findings in the PANDAS subgroup. AB - BACKGROUND: Sydenham's chorea is the neurologic manifestation of rheumatic fever and is a diagnosis of exclusion requiring only the presence of frank chorea in the absence of another neurologic disorder. Two thirds of children with Sydenham's chorea also have rheumatic carditis (pathologic mitral valve regurgitation). Although there are similar neuropsychiatric symptoms and preceding group A beta-hemolytic streptococcal infection associated with both Sydenham's chorea and the PANDAS (pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections) subgroup, it is unknown whether patients in the PANDAS subgroup have any cardiac involvement. METHODS: Sixty children meeting the criteria for PANDAS were entered into protocols at National Institute of Mental Health between 1993 and 2002. Doppler and 2-dimensional echocardiograms were performed on these subjects to assess valvular heart disease. RESULTS: Of these 60 children, no echocardiographic evidence of significant mitral or aortic valve regurgitation was found. One patient was found to have mild mitral regurgitation, and all patients had normal left atrial size and normal left ventricular size and function. Follow-up echocardiograms on 20 children showed no significant valvular regurgitation. CONCLUSION: The evidence of a clear lack of rheumatic carditis in these children supports the hypothesis that PANDAS is a distinct neuropsychiatric diagnosis separate from Sydenham's chorea. PMID- 15545619 TI - HIV infection and zidovudine use in childbearing women. AB - OBJECTIVE: The risk of vertical HIV transmission from mother to child has been shown to be markedly decreased through HIV education, counseling, testing, and zidovudine (ZDV) use. The US Public Health Service published guidelines in 1994 for the use of ZDV on the basis of results of the AIDS Clinical Trials Group Protocol, a multicenter clinical trial of ZDV. The Public Health Service followed these guidelines with recommendations for routine HIV counseling and testing with informed consent for all pregnant women. New Jersey adopted these guidelines as the standard of care and created a program implementing the standard in all hospitals with maternity units. The purpose of this report was to study the trends in the rate of HIV infection in childbearing women over the past decade and to follow patterns of use of ZDV in the HIV-positive women, as a marker for the success of New Jersey's policy and program to reduce mother-to-child transmission. METHODS: Since 1988 in New Jersey, blood from heel-stick filter papers has been tested for the presence of HIV antibody through anonymous, unlinked surveys. Excess blood from screening for inborn errors of metabolism for all infants who were born in the state from July through September of each year was tested using a Food and Drug Administration-approved HIV-1 and Western blot test. Age, race, and ethnicity were recorded, as well as the results from the HIV 1 and Western blot tests. Since 1994, specimens confirmed to be HIV-1 positive by Western blot test were tested for the presence of ZDV, and the results were recorded. RESULTS: The number of New Jersey women included in the study for the period 1990 through 2002 numbered 372305. The percentage of childbearing women who tested positive for HIV declined by 55% during the period, although the declines were not uniform in all subgroups. In the early 1990s, women who were <30 years old had higher infection rates than older women, but this has not been a consistent pattern during the period. Younger women again had a higher rate in 2002. When HIV-positive rates are examined by 5-year age groups, the declines are dramatic for younger women. The rate per 100 women 20 through 24 years decreased from 0.46 per 100 tested women in 1990 to 0.29 in 2002 and for women 25 through 29 years from 0.51 per 100 in 1990 to 0.25 in 2002. The rate for women 30 through 34 years of age declined from 0.54 in 1990 to 0.13 in 2002. During the same time period, the rate per 100 tested women 35 though 39 years of age increased from 0.23 to 0.33. Black non-Hispanic women who give birth to live infants have the highest HIV-positive rates, followed by Hispanic women and white non-Hispanic women. In 2002, this rate was 0.74 per 100 in black non-Hispanic women, 0.22 per 100 in Hispanic women, and 0.08 in white non-Hispanic women. Although major disparities continue, the infection rate in black non-Hispanic women demonstrated the greatest decrease during the period, followed by the decline among Hispanic women. The use of ZDV in HIV-positive women increased dramatically during the period, from 13.3% in 1994, when it was first tested in New Jersey, to an all time high of 88.5% in 2002. CONCLUSIONS: Reducing perinatal HIV transmission is a priority for the New Jersey Department of Health and Senior Services. Reducing perinatal transmission can be accomplished by reducing the number of infants who are exposed perinatally or decreasing the percentage of exposed infants for whom transmission occurs or both. The decrease in prevalence of HIV-positive status in childbearing women is in opposition to an overall increasing trend in prevalence rates. This decrease is thought to be attributable in part to the positive impact of numerous education and prevention programs but may also be the result of a voluntary decision on the part of HIV-infected women not to become pregnant or not to carry to term. In addition, the cohort of women who became infected in the early years of the epidemic may be aging out of their childbearing years, may have more advanced disease with a concomitant difficulty with fertility and carrying to term, or may have died. In New Jersey, a greater proportion of women with newly diagnosed HIV disease are past their childbearing years as compared with earlier years. Increased use of ZDV is thought to be attributable to several factors: dissemination of information to health care providers via continuing medical education activities; dissemination of information to the public, in particular to women; outreach via community-based organizations; and New Jersey Department of Health and Senior Service regulations and policies for mandatory counseling and voluntary testing of all pregnant women. A recent addition to New Jersey's comprehensive program to decrease perinatal transmission occurred in 2002 with dissemination to hospitals of the department's standard of care for women who present in labor with unknown HIV status. Physicians, nurses, and hospitals play vital roles in preventing vertical transmission of HIV by providing preconception and postconception counseling, testing with consent of pregnant women, and treatment for HIV-positive mothers, including administration of ZDV. This study not only provides an estimate of the prevalence of HIV infection in the population of childbearing women but also provides a means of examining the vertical transmission of HIV infection from mother to child. Continued research on this subpopulation as well as on other groups will provide additional knowledge to help in the overall goal of reducing HIV prevalence. PMID- 15545620 TI - Bedside limited echocardiography by the emergency physician is accurate during evaluation of the critically ill patient. AB - OBJECTIVE: Echocardiography can be a rapid, noninvasive, objective tool in the assessment of ventricular function and preload during resuscitation of a critically ill or injured child. We sought to determine the accuracy of bedside limited echocardiography by the emergency physician (BLEEP) in estimation of (1) left ventricular function (LVF) and (2) inferior vena cava (IVC) volume, as an indirect measure of preload. METHODS: We conducted a prospective observational study of a convenience sample of patients who were admitted to our intensive care unit. All patients underwent BLEEP followed by an independent formal echocardiogram by an experienced pediatric echocardiography provider (PEP). IVC volume was assessed by measurement of the maximal diameter of the IVC. LVF was determined by calculating shortening fraction (SF) using M-mode measurements on the parasternal short-axis view at the level of the papillary muscle. An independent blinded pediatric cardiologist reviewed all images for accuracy and quality. Estimates of SF obtained on the BLEEP examination were compared with those obtained by the PEP. RESULTS: Thirty-one patients were enrolled. The mean age was 5.1 years (range: 23 days-16 years); 48.4% (15 of 31) were girls; 58.1% (18 of 31) were on mechanical ventilatory support at the time of their study. There was good agreement between the emergency physician (EP) and the PEP for estimation of SF (r = 0.78). The mean difference in the estimate of SF between the providers was 4.4% (95% confidence interval: 1.6%-7.2%). This difference in estimate of SF was statistically significant. Similarly, there was good agreement between the EP and the PEP for estimation of IVC volume (r = 0.8). The mean difference in the estimate of IVC diameter by the PEP and the EP was 0.068 mm (95% confidence interval: -0.16 to 0.025 mm). This difference was not statistically significant. CONCLUSIONS: Our study suggests that PEP sonographers are capable of obtaining images that permit accurate assessment of LVF and IVC volume. BLEEP can be performed with focused training and oversight by a pediatric cardiologist. PMID- 15545621 TI - The natural history of type B Niemann-Pick disease: results from a 10-year longitudinal study. AB - OBJECTIVES: Type B Niemann-Pick disease (NPD-B) caused by acid sphingomyelinase deficiency is a rare, autosomal recessive, lysosomal storage disorder with a broad range of disease severity. The objectives of this study were to document the natural history of the disease in a large, clinically heterogeneous patient population that was followed for a period of 10 years and to determine how genotype influences phenotype. METHODS: Twenty-nine patients with NPD-B had serial evaluations at least 9 months apart. Organ volumes, hematologic indices, lipid concentrations, pulmonary function, and hepatic activity were studied, and individual phenotypic severity was compared with genotype. RESULTS: All patients with intact spleens had splenomegaly (mean value: 12.7 multiples of normal [MN]; range: 4.5-27.3 MN), and all but 1 had hepatomegaly (mean volume: 1.91 MN; range: 0.93-3.21 MN). At initial visit, 39% had thrombocytopenia and 3% had leukopenia. At final visit, the percentages increased to 54% and 34%, respectively. Mean annual decreases in platelet count and leukocyte count were 7 x 10(3) and 0.2 x 10(3) per mm3, respectively. The typical atherogenic lipid profile was worse in older patients. A total of 69% of patients had low diffusion capacity for carbon monoxide, and more than one third had low forced expiratory volume in 1 second, forced vital capacity, and forced expiratory volume in 1 second/forced vital capacity at initial visit. All measurements of pulmonary function showed a gradual deterioration over time. Liver dysfunction was characterized by stable elevation of hepatic transaminases and bilirubin. Homozygotes for DeltaR608, P323A, and P330R had milder disease than patients with all other genotypes. CONCLUSIONS: The natural history of NPD-B is characterized by hepatosplenomegaly with progressive hypersplenism, worsening atherogenic lipid profile, gradual deterioration in pulmonary function, and stable liver dysfunction. PMID- 15545622 TI - siRNAs targeting an intronic transposon in the regulation of natural flowering behavior in Arabidopsis. AB - Allelic variation in FLOWERING LOCUS C (FLC), a central repressor of flowering, contributes to natural differences in flowering behavior among Arabidopsis accessions. The weak nature of the FLC allele in the Ler accession is due to low levels of FLC RNA resulting, through an unknown mechanism, from a transposable element inserted in an intron of FLC. Here we show that the transposable element renders FLC-Ler subject to repressive chromatin modifications mediated by short interfering RNAs generated from homologous transposable elements in the genome. Our studies have general implications for the role of transposable elements in eukaryotic gene expression and evolution. PMID- 15545623 TI - JNK potentiates TNF-stimulated necrosis by increasing the production of cytotoxic reactive oxygen species. AB - The c-Jun NH(2)-terminal kinase (JNK) has been implicated in both cell death and survival responses to different stimuli. Here we reexamine the function of JNK in tumor necrosis factor (TNF)-stimulated cell death using fibroblasts isolated from wild-type, Mkk4(-/-) Mkk7(-/-), and Jnk1(-/-) Jnk2(-/-) mice. We demonstrate that JNK can act to suppress TNF-stimulated apoptosis. However, we find that JNK can also potentiate TNF-stimulated necrosis by increasing the production of reactive oxygen species (ROS). Together, these data indicate that JNK can shift the balance of TNF-stimulated cell death from apoptosis to necrosis. Increased necrosis may represent a contributing factor in stress-induced inflammatory responses mediated by JNK. PMID- 15545624 TI - Identification of a Drosophila Myb-E2F2/RBF transcriptional repressor complex. AB - The Drosophila Myb complex has roles in both activating and repressing developmentally regulated DNA replication. To further understand biochemically the functions of the Myb complex, we fractionated Drosophila embryo extracts relying upon affinity chromatography. We found that E2F2, DP, RBF1, RBF2, and the Drosophila homolog of LIN-52, a class B synthetic multivulva (synMuv) protein, copurify with the Myb complex components to form the Myb-MuvB complex. In addition, we found that the transcriptional repressor protein, lethal (3) malignant brain tumor protein, L(3)MBT, and the histone deacetylase, Rpd3, associated with the Myb-MuvB complex. Members of the Myb-MuvB complex were localized to promoters and were shown to corepress transcription of developmentally regulated genes. These and other data now link together the Myb and E2F2 complexes in higher-order assembly to specific chromosomal sites for the regulation of transcription. PMID- 15545625 TI - Regulation of mTOR function in response to hypoxia by REDD1 and the TSC1/TSC2 tumor suppressor complex. AB - Mammalian target of rapamycin (mTOR) is a central regulator of protein synthesis whose activity is modulated by a variety of signals. Energy depletion and hypoxia result in mTOR inhibition. While energy depletion inhibits mTOR through a process involving the activation of AMP-activated protein kinase (AMPK) by LKB1 and subsequent phosphorylation of TSC2, the mechanism of mTOR inhibition by hypoxia is not known. Here we show that mTOR inhibition by hypoxia requires the TSC1/TSC2 tumor suppressor complex and the hypoxia-inducible gene REDD1/RTP801. Disruption of the TSC1/TSC2 complex through loss of TSC1 or TSC2 blocks the effects of hypoxia on mTOR, as measured by changes in the mTOR targets S6K and 4E-BP1, and results in abnormal accumulation of Hypoxia-inducible factor (HIF). In contrast to energy depletion, mTOR inhibition by hypoxia does not require AMPK or LKB1. Down-regulation of mTOR activity by hypoxia requires de novo mRNA synthesis and correlates with increased expression of the hypoxia-inducible REDD1 gene. Disruption of REDD1 abrogates the hypoxia-induced inhibition of mTOR, and REDD1 overexpression is sufficient to down-regulate S6K phosphorylation in a TSC1/TSC2 dependent manner. Inhibition of mTOR function by hypoxia is likely to be important for tumor suppression as TSC2-deficient cells maintain abnormally high levels of cell proliferation under hypoxia. PMID- 15545626 TI - The hypoxia-induced paralogs Scylla and Charybdis inhibit growth by down regulating S6K activity upstream of TSC in Drosophila. AB - Diverse extrinsic and intrinsic cues must be integrated within a developing organism to ensure appropriate growth at the cellular and organismal level. In Drosophila, the insulin receptor/TOR/S6K signaling network plays a fundamental role in the control of metabolism and cell growth. Here we show that scylla and charybdis, two homologous genes identified as growth suppressors in an EP (enhancer/promoter) overexpression screen, act as negative regulators of growth. The simultaneous loss of both genes generates flies that are more susceptible to reduced oxygen concentrations (hypoxia) and that show mild overgrowth phenotypes. Conversely, scylla or charybdis overactivation reduces growth. Growth inhibition is associated with a reduction in S6K but not PKB/Akt activity. Together, genetic and biochemical analysis places Scylla/Charybdis downstream of PKB and upstream of TSC. Furthermore, we show that scylla and charybdis are induced under hypoxic conditions and that scylla is a target of Drosophila HIF-1 (hypoxia-inducible factor-1) like its mammalian counterpart RTP801/REDD1, thus establishing a potential cross-talk between growth and oxygen sensing. PMID- 15545627 TI - Living with or without cyclins and cyclin-dependent kinases. AB - Entry into, progression through, and exit from the G1 phase of the mammalian cell cycle in response to extracellular mitogenic cues are presumed to be governed by cyclin-dependent kinases (Cdks) regulated by the D-type and E-type cyclins. Studies performed over more than a decade have supported the view that these holoenzymes are important, if not required, for these processes. However, recent experiments in which the genes encoding all three D-type cyclins, the two E-type cyclins, cyclin D-dependent Cdk4 and Cdk6, or cyclin E-dependent Cdk2 have been disrupted in the mouse germ line have revealed that much of fetal development occurs normally in their absence. Thus, none of these genes is strictly essential for cell cycle progression. To what extent is the prevailing dogma incorrect, and how can the recent findings be reconciled with past work? PMID- 15545628 TI - WNT signaling, in synergy with T/TBX6, controls Notch signaling by regulating Dll1 expression in the presomitic mesoderm of mouse embryos. AB - Notch signaling in the presomitic mesoderm (psm) is critical for somite formation and patterning. Here, we show that WNT signals regulate transcription of the Notch ligand Dll1 in the tailbud and psm. LEF/TCF factors cooperate with TBX6 to activate transcription from the Dll1 promoter in vitro. Mutating either T or LEF/TCF sites in the Dll1 promoter abolishes reporter gene expression in vitro as well as in the tail bud and psm of transgenic embryos. Our results indicate that WNT activity, in synergy with TBX6, regulates Dll1 transcription and thereby controls Notch activity, somite formation, and patterning. PMID- 15545629 TI - LEF1-mediated regulation of Delta-like1 links Wnt and Notch signaling in somitogenesis. AB - Wnt signaling, which is mediated by LEF1/TCF transcription factors, has been placed upstream of the Notch pathway in vertebrate somitogenesis. Here, we examine the molecular basis for this presumed hierarchy and show that a targeted mutation of Lef1, which abrogates LEF1 function and impairs the activity of coexpressed TCF factors, affects the patterning of somites and the expression of components of the Notch pathway. LEF1 was found to bind multiple sites in the Dll1 promoter in vitro and in vivo. Moreover, mutations of LEF1-binding sites in the Dll1 promoter impair expression of a Dll1-LacZ transgene in the presomitic mesoderm. Finally, the induced expression of LEF1-beta-catenin activates the expression of endogenous Dll1 in fibroblastic cells. Thus, Wnt signaling can affect the Notch pathway by a LEF1-mediated regulation of Dll1. PMID- 15545630 TI - MIM/BEG4, a Sonic hedgehog-responsive gene that potentiates Gli-dependent transcription. AB - Sonic hedgehog (Shh) signaling plays a critical role during development and carcinogenesis. While Gli family members govern the transcriptional output of Shh signaling, little is known how Gli-mediated transcriptional activity is regulated. Here we identify the actin-binding protein Missing in Metastasis (MIM) as a new Shh-responsive gene. Together, Gli1 and MIM recapitulate Shh-mediated epidermal proliferation and invasion in regenerated human skin. MIM is part of a Gli/Suppressor of Fused complex and potentiates Gli-dependent transcription using domains distinct from those used for monomeric actin binding. These data define MIM as both a Shh-responsive gene and a new member of the pathway that modulates Gli responses during growth and tumorigenesis. PMID- 15545631 TI - Notch3 is required for arterial identity and maturation of vascular smooth muscle cells. AB - Formation of a fully functional artery proceeds through a multistep process. Here we show that Notch3 is required to generate functional arteries in mice by regulating arterial differentiation and maturation of vascular smooth muscle cells (vSMC). In adult Notch3-/- mice distal arteries exhibit structural defects and arterial myogenic responses are defective. The postnatal maturation stage of vSMC is deficient in Notch3-/- mice. We further show that Notch3 is required for arterial specification of vSMC but not of endothelial cells. Our data reveal Notch3 to be the first cell-autonomous regulator of arterial differentiation and maturation of vSMC. PMID- 15545632 TI - c-Myc controls the balance between hematopoietic stem cell self-renewal and differentiation. AB - The activity of adult stem cells is essential to replenish mature cells constantly lost due to normal tissue turnover. By a poorly understood mechanism, stem cells are maintained through self-renewal while concomitantly producing differentiated progeny. Here, we provide genetic evidence for an unexpected function of the c-Myc protein in the homeostasis of hematopoietic stem cells (HSCs). Conditional elimination of c-Myc activity in the bone marrow (BM) results in severe cytopenia and accumulation of HSCs in situ. Mutant HSCs self-renew and accumulate due to their failure to initiate normal stem cell differentiation. Impaired differentiation of c-Myc-deficient HSCs is linked to their localization in the differentiation preventative BM niche environment, and correlates with up regulation of N-cadherin and a number of adhesion receptors, suggesting that release of HSCs from the stem cell niche requires c-Myc activity. Accordingly, enforced c-Myc expression in HSCs represses N-cadherin and integrins leading to loss of self-renewal activity at the expense of differentiation. Endogenous c-Myc is differentially expressed and induced upon differentiation of long-term HSCs. Collectively, our data indicate that c-Myc controls the balance between stem cell self-renewal and differentiation, presumably by regulating the interaction between HSCs and their niche. PMID- 15545633 TI - Cold shock and regulation of surface protein trafficking convey sensitization to inducers of stage differentiation in Trypanosoma brucei. AB - Transmission of a protozoan parasite from a vertebrate to invertebrate host is accompanied by cellular differentiation. The signals from the environment that trigger the process are poorly understood. The model parasite Trypanosoma brucei proliferates in the mammalian bloodstream and in the tsetse fly. On ingestion by the tsetse, the trypanosome undergoes a rapid differentiation that is marked by replacement of the variant surface glycoprotein (VSG) coat with GPI-anchored EP and GPEET procyclins. Here we show that a cold shock of DeltaT > 15 degrees C is sufficient to reversibly induce high-level expression of the insect stage specific EP gene in the mammalian bloodstream stages of T. brucei. The 3'-UTR of the EP mRNA is necessary and sufficient for the increased expression. During cold shock, EP protein accumulates in the endosomal compartment in the proliferating, slender, bloodstream stage, whereas the EP is present on the plasma membrane in the quiescent, stumpy, bloodstream stage. Thus, there is a novel developmentally regulated cell surface access control mechanism for a GPI-anchored protein. In addition to inducing EP expression, cold shock results in the acquisition of sensitivity to micromolar concentrations of cis-aconitate and citrate by stumpy but not slender bloodstream forms. The cis-aconitate and citrate commit stumpy bloodstream cells to differentiation to the procyclic stage along with rapid initial proliferation. We propose a hierarchical model of three events that regulate differentiation after transmission to the tsetse: sensing the temperature change, surface access of a putative receptor, and sensing of a chemical cue. PMID- 15545634 TI - A chaperone network controls the heat shock response in E. coli. AB - The heat shock response controls levels of chaperones and proteases to ensure a proper cellular environment for protein folding. In Escherichia coli, this response is mediated by the bacterial-specific transcription factor, sigma32. The DnaK chaperone machine regulates both the amount and activity of sigma32, thereby coupling sigma32 function to the cellular protein folding state. In this manuscript, we analyze the ability of other major chaperones in E. coli to regulate sigma32, and we demonstrate that the GroEL/S chaperonin is an additional regulator of sigma32. We show that increasing the level of GroEL/S leads to a decrease in sigma32 activity in vivo and this effect can be eliminated by co overexpression of a GroEL/S-specific substrate. We also show that depletion of GroEL/S in vivo leads to up-regulation of sigma32 by increasing the level of sigma32. In addition, we show that changing the levels of GroEL/S during stress conditions leads to measurable changes in the heat shock response. Using purified proteins, we show that that GroEL binds to sigma32 and decreases sigma32 dependent transcription in vitro, suggesting that this regulation is direct. We discuss why using a chaperone network to regulate sigma32 results in a more sensitive and accurate detection of the protein folding environment. PMID- 15545635 TI - Vascular endothelial growth factor controls neuronal migration and cooperates with Sema3A to pattern distinct compartments of the facial nerve. AB - Developing neurons accurately position their somata within the neural tube to make contact with appropriate neighbors and project axons to their preferred targets. Taking advantage of a collection of genetically engineered mouse mutants, we now demonstrate that the behavior of somata and axons of the facial nerve is regulated independently by two secreted ligands for the transmembrane receptor neuropilin 1 (Nrp1), the semaphorin Sema3A and the VEGF164 isoform of Vascular Endothelial Growth Factor. Although Sema3A is known to control the guidance of facial nerve axons, we now show that it is not required for the pathfinding of their somata. Vice versa, we find that VEGF164 is not required for axon guidance of facial motor neurons, but is essential for the correct migration of their somata. These observations demonstrate, for the first time, that VEGF contributes to neuronal patterning in vivo, and that different compartments of one cell can be co-ordinately patterned by structurally distinct ligands for a shared receptor. PMID- 15545636 TI - Roles of Rad23 protein in yeast nucleotide excision repair. AB - Nucleotide excision repair (NER) removes many different types of DNA lesions. Most NER proteins are indispensable for repair. In contrast, the yeast Rad23 represents a class of accessory NER proteins, without which NER activity is reduced but not eliminated. In mammals, the complex of HR23B (Rad23 homolog) and XPC (yeast Rad4 homolog) has been suggested to function in the damage recognition step of NER. However, the precise function of Rad23 or HR23B in NER remains unknown. Recently, it was suggested that the primary function of RAD23 protein in NER is its stabilization of XPC protein. Here, we tested the significance of Rad23-mediated Rad4 stabilization in NER, and analyzed the repair and biochemical activities of purified yeast Rad23 protein. Cellular Rad4 was indeed stabilized by Rad23 in the absence of DNA damage. Persistent overexpression of Rad4 in rad23 mutant cells, however, largely failed to complement the ultraviolet sensitivity of the mutant. Consistently, deficient NER in rad23 mutant cell extracts could not be complemented by purified Rad4 protein in vitro. In contrast, partial complementation was observed with purified Rad23 protein. Specific complementation to the level of wild-type repair was achieved by adding purified Rad23 together with small amounts of Rad4 protein to rad23 mutant cell extracts. Purified Rad23 protein was unable to bind to DNA, but stimulated the binding activity of purified Rad4 protein to N-acetyl-2-aminofluorene-damaged DNA. These results support two roles of Rad23 protein in NER: (i) its direct participation in the repair biochemistry, possibly due to its stimulatory activity on Rad4 mediated damage binding/recognition; and (ii) its stabilization of cellular Rad4 protein. PMID- 15545637 TI - RNA interference using boranophosphate siRNAs: structure-activity relationships. AB - In RNA interference (RNAi), short double-stranded RNA (known as siRNA) inhibits expression from homologous genes. Clinical or pre-clinical use of siRNAs is likely to require stabilizing modifications because of the prevalence of intracellular and extracellular nucleases. In order to examine the effect of modification on siRNA efficacy and stability, we developed a new method for synthesizing stereoregular boranophosphate siRNAs. This work demonstrates that boranophosphate siRNAs are consistently more effective than siRNAs with the widely used phosphorothioate modification. Furthermore, boranophosphate siRNAs are frequently more active than native siRNA if the center of the antisense strand is not modified. Boranophosphate modification also increases siRNA potency. The finding that boranophosphate siRNAs are at least ten times more nuclease resistant than unmodified siRNAs may explain some of the positive effects of boranophosphate modification. The biochemical properties of boranophosphate siRNAs make them promising candidates for an RNAi-based therapeutic. PMID- 15545638 TI - A natural meiotic DNA break site in Schizosaccharomyces pombe is a hotspot of gene conversion, highly associated with crossing over. AB - In Schizosaccharomyces pombe, meiosis-specific DNA breaks that initiate recombination are observed at prominent but widely separated sites. We investigated the relationship between breakage and recombination at one of these sites, the mbs1 locus on chromosome I. Breaks corresponding to 10% of chromatids were mapped to four clusters spread over a 2.1-kb region. Gene conversion of markers within the clusters occurred in 11% of tetrads (3% of meiotic chromatids), making mbs1 a conversion hotspot when compared to other fission yeast markers. Approximately 80% of these conversions were associated with crossing over of flanking markers, suggesting a strong bias in meiotic break repair toward the generation of crossovers. This bias was observed in conversion events at three other loci, ade6, ade7, and ura1. A total of 50-80% of all crossovers seen in a 90-kb region flanking mbs1 occurred in a 4.8-kb interval containing the break sites. Thus, mbs1 is also a hotspot of crossing over, with breakage at mbs1 generating most of the crossovers in the 90-kb interval. Neither Rec12 (Spo11 ortholog) nor I-SceI-induced breakage at mbs1 was significantly associated with crossing over in an apparently break-free interval >25 kb away. Possible mechanisms for generating crossovers in such break-free intervals are discussed. PMID- 15545639 TI - Hsp70 chaperones as modulators of prion life cycle: novel effects of Ssa and Ssb on the Saccharomyces cerevisiae prion [PSI+]. AB - [PSI(+)] is a prion isoform of the yeast release factor Sup35. In some assays, the cytosolic chaperones Ssa1 and Ssb1/2 of the Hsp70 family were previously shown to exhibit "pro-[PSI(+)]" and "anti-[PSI(+)]" effects, respectively. Here, it is demonstrated for the first time that excess Ssa1 increases de novo formation of [PSI(+)] and that pro-[PSI(+)] effects of Ssa1 are shared by all other Ssa proteins. Experiments with chimeric constructs show that the peptide binding domain is a major determinant of differences in the effects of Ssa and Ssb proteins on [PSI(+)]. Surprisingly, overproduction of either chaperone increases loss of [PSI(+)] when Sup35 is simultaneously overproduced. Excess Ssa increases both the average size of prion polymers and the proportion of monomeric Sup35 protein. Both in vivo and in vitro experiments uncover direct physical interactions between Sup35 and Hsp70 proteins. The proposed model postulates that Ssa stimulates prion formation and polymer growth by stabilizing misfolded proteins, which serve as substrates for prion conversion. In the case of very large prion aggregates, further increase in size may lead to the loss of prion activity. In contrast, Ssb either stimulates refolding into nonprion conformation or targets misfolded proteins for degradation, in this way counteracting prion formation and propagation. PMID- 15545640 TI - Bonus, a Drosophila TIF1 homolog, is a chromatin-associated protein that acts as a modifier of position-effect variegation. AB - Bonus, a Drosophila TIF1 homolog, is a nuclear receptor cofactor required for viability, molting, and numerous morphological events. Here we establish a role for Bonus in the modulation of chromatin structure. We show that weak loss-of function alleles of bonus have a more deleterious effect on males than on females. This male-enhanced lethality is not due to a defect in dosage compensation or somatic sex differentiation, but to the presence of the Y chromosome. Additionally, we show that bonus acts as both an enhancer and a suppressor of position-effect variegation. By immunostaining, we demonstrate that Bonus is associated with both interphase and prophase chromosomes and through chromatin immunoprecipitation show that two of these sites correspond to the histone gene cluster and the Stellate locus. PMID- 15545641 TI - Evidence for abundant slightly deleterious polymorphisms in bacterial populations. AB - The nearly neutral theory of molecular evolution predicts that slightly deleterious mutations subject to purifying selection are widespread in natural populations, particularly those of large effective population size. To test this hypothesis, the standardized difference between pairwise nucleotide difference and number of segregation sites (corrected for number of sequences) was estimated for 149 population data sets from 84 species of bacteria. This quantity (Tajima's D-statistic) was estimated separately for synonymous (D(syn)) and nonsynonymous (D(non)) polymorphisms. D(syn) was positive in 70% of data sets, and the overall median D(syn) (0.873) was positive. By contrast D(non) was negative in 68% of data sets, and the overall median D(non) (-0.656) was negative. The preponderance of negative values of D(non) is evidence that there are widespread rare nonsynonymous polymorphisms in the process of being eliminated by purifying selection, as predicted to occur in populations with large effective size by the nearly neutral theory. The major exceptions to this trend were seen among surface proteins, particularly those of bacteria parasitic on vertebrates, which included a number of cases of polymorphisms apparently maintained by balancing selection. PMID- 15545642 TI - Missense mutations that inactivate the Aspergillus nidulans nrtA gene encoding a high-affinity nitrate transporter. AB - The transport of nitrate into prokaryotic and eukaryotic cells, of considerable interest to agriculture, ecology, and human health, is carried out by members of a distinct cluster of proteins within the major facilitator superfamily. To obtain structure/function information on this important class of nitrate permeases, a collection of chemically induced mutations in the nrtA gene encoding a 12-transmembrane domain, high-affinity nitrate transporter from the eukaryote Aspergillus nidulans was isolated and characterized. This mutational analysis, coupled with protein alignments, demonstrates the utility of the approach to predicting peptide motifs and individual residues important for the movement of nitrate across the membrane. These include the highly conserved nitrate signature motif (residues 166-173) in Tm 5, the conserved charged residues Arg87 (Tm 2) and Arg368 (Tm 8), as well as the aromatic residue Phe47 (Tm 1), all within transmembrane helices. No mutations were observed in the large central loop (Lp 6/7) between Tm 6 and Tm 7. Finally, the study of a strain with a conversion of Trp481 (Tm 12) to a stop codon suggests that all 12 transmembrane domains and/or the C-terminal tail are required for membrane insertion and/or stability of NrtA. PMID- 15545644 TI - Protecting haploid polymorphisms in temporally variable environments. AB - Analysis of a continuous-time model shows that a protected polymorphism can arise in a haploid population subject to temporal fluctuations in selection. The requirements are that population size is regulated in a density-dependent manner and that an allele's arithmetic mean relative growth rate is greater than one when rare and that its harmonic mean relative growth rate is less than one when common. There is no requirement that relative growth rate be frequency dependent. Comparisons with discrete-time models show that the standard formalism used by population genetics ignores forced changes in generation time as rare advantageous alleles sweep into a population. In temporally variable environments, frequency-dependent changes in generation times tend to counteract these invasions. Such changes can prevent fixation and protect polymorphisms. PMID- 15545643 TI - An active transposable element, Herves, from the African malaria mosquito Anopheles gambiae. AB - Transposable elements have proven to be invaluable tools for genetically manipulating a wide variety of plants, animals, and microbes. Some have suggested that they could be used to spread desirable genes, such as refractoriness to Plasmodium infection, through target populations of Anopheles gambiae, thereby disabling the mosquito's ability to transmit malaria. To achieve this, a transposon must remain mobile and intact after the initial introduction into the genome. Endogenous, active class II transposable elements from An. gambiae have not been exploited as gene vectors/drivers because none have been isolated. We report the discovery of an active class II transposable element, Herves, from the mosquito An. gambiae. Herves is a member of a distinct subfamily of hAT elements that includes the hopper-we element from Bactrocera dorsalis and B. cucurbitae. Herves was transpositionally active in mobility assays performed in Drosophila melanogaster S2 cells and developing embryos and was used as a germ-line transformation vector in D. melanogaster. Herves displays an altered target-site preference from the distantly related hAT elements, Hermes and hobo. Herves is also present in An. arabiensis and An. merus with copy numbers similar to that found in An. gambiae. Preliminary data from an East African population are consistent with the element being transpositionally active in mosquitoes. PMID- 15545645 TI - RNA silencing in Aspergillus nidulans is independent of RNA-dependent RNA polymerases. AB - The versatility of RNA-dependent RNA polymerases (RDRPs) in eukaryotic gene silencing is perhaps best illustrated in the kingdom Fungi. Biochemical and genetic studies of Schizosaccharomyces pombe and Neurospora crassa show that these types of enzymes are involved in a number of fundamental gene-silencing processes, including heterochromatin regulation and RNA silencing in S. pombe and meiotic silencing and RNA silencing in N. crassa. Here we show that Aspergillus nidulans, another model fungus, does not require an RDRP for inverted repeat transgene (IRT)-induced RNA silencing. However, RDRP requirements may vary within the Aspergillus genus as genomic analysis indicates that A. nidulans, but not A. fumigatus or A. oryzae, has lost a QDE-1 ortholog, an RDRP associated with RNA silencing in N. crassa. We also provide evidence suggesting that 5' --> 3' transitive RNA silencing is not a significant aspect of A. nidulans IRT-RNA silencing. These results indicate a lack of conserved kingdom-wide requirements for RDRPs in fungal RNA silencing. PMID- 15545646 TI - Meiotic recombination in Drosophila females depends on chromosome continuity between genetically defined boundaries. AB - In the pairing-site model, specialized regions on each chromosome function to establish meiotic homolog pairing. Analysis of these sites could provide insights into the mechanism used by Drosophila females to form a synaptonemal complex (SC) in the absence of meiotic recombination. These specialized sites were first established on the X chromosome by noting that there were barriers to crossover suppression caused by translocation heterozygotes. These sites were genetically mapped and proposed to be pairing sites. By comparing the cytological breakpoints of third chromosome translocations to their patterns of crossover suppression, we have mapped two sites on chromosome 3R. We have performed experiments to determine if these sites have a role in meiotic homolog pairing and the initiation of recombination. Translocation heterozygotes exhibit reduced gene conversion within the crossover-suppressed region, consistent with an effect on the initiation of meiotic recombination. To determine if homolog pairing is disrupted in translocation heterozygotes, we used fluorescent in situ hybridization to measure the extent of homolog pairing. In wild-type oocytes, homologs are paired along their entire lengths prior to accumulation of the SC protein C(3)G. Surprisingly, translocation heterozygotes exhibited homolog pairing similar to wild type within the crossover-suppressed regions. This result contrasted with our observations of c(3)G mutant females, which were found to be defective in pairing. We propose that each Drosophila chromosome is divided into several domains by specialized sites. These sites are not required for homolog pairing. Instead, the initiation of meiotic recombination requires continuity of the meiotic chromosome structure within each of these domains. PMID- 15545647 TI - The genomes of recombinant inbred lines. AB - Recombinant inbred lines (RILs) can serve as powerful tools for genetic mapping. Recently, members of the Complex Trait Consortium proposed the development of a large panel of eight-way RILs in the mouse, derived from eight genetically diverse parental strains. Such a panel would be a valuable community resource. The use of such eight-way RILs will require a detailed understanding of the relationship between alleles at linked loci on an RI chromosome. We extend the work of Haldane and Waddington on two-way RILs and describe the map expansion, clustering of breakpoints, and other features of the genomes of multiple-strain RILs as a function of the level of crossover interference in meiosis. PMID- 15545648 TI - Genetic factors that regulate the attenuation of the general stress response of yeast. AB - The general stress response of yeast involves the induction of approximately 200 genes in response to any one of several stresses. These genes are activated by Msn2 and repressed by the Srb10 kinase, a member of the mediator complex. Normally, Msn2 is exported from the nucleus, and Srb10 represses STRE gene expression. Under stress, Msn2 relocalizes to the nucleus and, with the relief of Srb10 repression, activates transcription. The stress response is rapid, but quickly attenuated. We show here that this attenuation is due to a nuclear dependent degradation of Msn2. Msn2 rapidly disappeared from cells after heat or osmotic shock. This disappearance was not due to a change in MSN2 RNA levels, which remain constant during stress. Pulse-chase experiments confirmed the stress dependent Msn2 degradation. The levels of Msn2 were significantly reduced in msn5 deletion cells that have been shown to constitutively retain Msn2 in the nucleus. The degradation was Srb10-dependent; Msn2 was not degraded in an srb10 deletion mutant. An Msn2 internal deletion mutant was insensitive to Srb10 repression, but was degraded by the Srb10-dependent mechanism. Thus, this mutation uncoupled Srb10 repression from degradation. PMID- 15545649 TI - Protein kinase A regulates constitutive expression of small heat-shock genes in an Msn2/4p-independent and Hsf1p-dependent manner in Saccharomyces cerevisiae. AB - Hsf1p, the heat-shock transcription factor from Saccharomyces cerevisiae, has a low level of constitutive transcriptional activity and is kept in this state through negative regulation. In an effort to understand this negative regulation, we developed a novel genetic selection that detects altered expression from the HSP26 promoter. Using this reporter strain, we identified mutations and dosage compensators in the Ras/cAMP signaling pathway that decrease cAMP levels and increase expression from the HSP26 promoter. In yeast, low cAMP levels reduce the catalytic activity of the cAMP-dependent kinase PKA. Previous studies had proposed that the stress response transcription factors Msn2p/4p, but not Hsf1p, are repressed by PKA. However, we found that reduction or elimination of PKA activity strongly derepresses transcription of the small heat-shock genes HSP26 and HSP12, even in the absence of MSN2/4. In a strain deleted for MSN2/4 and the PKA catalytic subunits, expression of HSP12 and HSP26 depends on HSF1 expression. Our findings indicate that Hsf1p functions downstream of PKA and suggest that PKA might be involved in negative regulation of Hsf1p activity. These results represent a major change in our understanding of how PKA signaling influences the heat-shock response and heat-shock protein expression. PMID- 15545650 TI - Interaction between the oxa1 and rmp1 genes modulates respiratory complex assembly and life span in Podospora anserina. AB - A causal link between deficiency of the cytochrome respiratory pathway and life span was previously shown in the filamentous fungus Podospora anserina. To gain more insight into the relationship between mitochondrial function and life span, we have constructed a strain carrying a thermosensitive mutation of the gene oxa1. OXA1 is a membrane protein conserved from bacteria to human. The mitochondrial OXA1 protein is involved in the assembly/insertion of several respiratory complexes. We show here that oxa1 is an essential gene in P. anserina. The oxa1(ts) mutant exhibits severe defects in the respiratory complexes I and IV, which are correlated with an increased life span, a strong induction of the alternative oxidase, and a reduction in ROS production. However, there is no causal link between alternative oxidase level and life span. We also show that in the oxa1(ts) mutant, the extent of the defects in complexes I and IV and the life-span increase depends on the essential gene rmp1. The RMP1 protein, whose function is still unknown, can be localized in the mitochondria and/or the cytosolic compartment, depending on the developmental stage. We propose that the RMP1 protein could be involved in the process of OXA1-dependent protein insertion. PMID- 15545651 TI - Lig4 and rad54 are required for repair of DNA double-strand breaks induced by P element excision in Drosophila. AB - Site-specific double-strand breaks (DSBs) were generated in the white gene located on the X chromosome of Drosophila by excision of the w(hd) P-element. To investigate the role of nonhomologous end joining (NHEJ) and homologous recombination (HR) in the repair of these breaks, the w(hd) P-element was mobilized in flies carrying mutant alleles of either lig4 or rad54. The survival of both lig4- and rad54-deficient males was reduced to 25% in comparison to the wild type, indicating that both NHEJ and HR are involved in the repair P-induced gaps in males. Survival of lig4-deficient females was not affected at all, implying that HR using the homologous chromosome as a template can partially compensate for the impaired NHEJ pathway. In rad54 mutant females survival was reduced to 70% after w(hd) excision. PCR analysis indicated that the undamaged homologous chromosome may compensate for the potential loss of the broken chromosome in rad54 mutant females after excision. Molecular analysis of the repair junctions revealed microhomology (2-8 bp)-dependent DSB repair in most products. In the absence of Lig4, the 8-bp target site duplication is used more frequently for repair. Our data indicate the presence of efficient alternative end-joining mechanisms, which partly depend on the presence of microhomology but do not require Lig4. PMID- 15545652 TI - The population structure of African cultivated rice oryza glaberrima (Steud.): evidence for elevated levels of linkage disequilibrium caused by admixture with O. sativa and ecological adaptation. AB - Genome-wide linkage disequilibrium (LD) was investigated for 198 accessions of Oryza glaberrima using 93 nuclear microsatellite markers. Significantly elevated levels of LD were detected, even among distantly located markers. Free recombination among loci at the population genetic level was shown (1) by a lack of decay in LD among markers on the same chromosome and (2) by a strictly increasing composite likelihood function for the recombination parameter. This suggested that the elevation in LD was due not to physical linkage but to other factors, such as population structure. A Bayesian clustering analysis confirmed this hypothesis, indicating that the sample of O. glaberrima in this study was subdivided into at least five cryptic subpopulations. Two of these subpopulations clustered with control samples of O. sativa, subspecies indica and japonica, indicating that some O. glaberrima accessions represent admixtures. The remaining three O. glaberrima subpopulations were significantly associated with specific combinations of phenotypic traits-possibly reflecting ecological adaptation to different growing environments. PMID- 15545653 TI - Patterns of selection on synonymous and nonsynonymous variants in Drosophila miranda. AB - We have investigated patterns of within-species polymorphism and between-species divergence for synonymous and nonsynonymous variants at a set of autosomal and X linked loci of Drosophila miranda. D. pseudoobscura and D. affinis were used for the between-species comparisons. The results suggest the action of purifying selection on nonsynonymous, polymorphic variants. Among synonymous polymorphisms, there is a significant excess of synonymous mutations from preferred to unpreferred codons and of GC to AT mutations. There was no excess of GC to AT mutations among polymorphisms at noncoding sites. This suggests that selection is acting to maintain the use of preferred codons. Indirect evidence suggests that biased gene conversion in favor of GC base pairs may also be operating. The joint intensity of selection and biased gene conversion, in terms of the product of effective population size and the sum of the selection and conversion coefficients, was estimated to be approximately 0.65. PMID- 15545654 TI - Gene genealogy and properties of test statistics of neutrality under population growth. AB - We consider the Wright-Fisher model with exponential population growth and investigate effects of population growth on the shape of genealogy and the distributions of several test statistics of neutrality. In the limiting case as the population grows rapidly, the rapid-growth-limit genealogy is characterized. We obtained approximate expressions for expectations and variances of test statistics in the rapid-growth-limit genealogy and star genealogy. The distributions in the star genealogy are narrower than those in the cases of the simulated and rapid-growth-limit genealogies. The expectations and variances of the test statistics are monotone decreasing functions of the time length of the expansion, and the higher power of R(2) against population growth is suggested to be due to their smaller variances rather than to change of the expectations. We also investigated by simulation how quickly the distributions of test statistics approach those of the rapid-growth-limit genealogy. PMID- 15545655 TI - Conserved locus-specific silencing functions of Schizosaccharomyces pombe sir2+. AB - In Schizosaccharomyces pombe, three genes, sir2(+), hst2(+), and hst4(+), encode members of the Sir2 family of conserved NAD(+)-dependent protein deacetylases. The S. pombe sir2(+) gene encodes a nuclear protein that is not essential for viability or for resistance to treatment with UV or a microtubule-destabilizing agent. However, sir2(+) is essential for full transcriptional silencing of centromeres, telomeres, and the cryptic mating-type loci. Chromatin immunoprecipitation results suggest that the Sir2 protein acts directly at these chromosomal regions. Enrichment of Sir2p at silenced regions does not require the HP1 homolog Swi6p; instead, Swi6-GFP localization to telomeres depends in part on Sir2p. The phenotype of sir2 swi6 double mutants supports a model whereby Sir2p functions prior to Swi6p at telomeres and the silent mating-type loci. However, Sir2p does not appear to be essential for the localization of Swi6p to centromeric foci. Cross-complementation experiments showed that the Saccharomyces cerevisiae SIR2 gene can function in place of S. pombe sir2(+), suggesting overlapping deacetylation substrates in both species. These results also suggest that, despite differences in most of the other molecules required, the two distantly related yeast species share a mechanism for targeting Sir2p homologs to silent chromatin. PMID- 15545656 TI - Genetic variation for total fitness in Drosophila melanogaster: complex yet replicable patterns. AB - The extent of genetic variation in fitness is a crucial issue in evolutionary biology and yet remains largely unresolved. In Drosophila melanogaster, we have devised a method that allows the net effects on fitness of heterozygous wild-type chromosomes to be measured, by competing them against two different "balancer" chromosomes. We have applied the method to a large sample of 40 wild-type third chromosomes and have measured fitnesses of nonlethal chromosomes as well as chromosomes bearing recessive lethals. The measurements were made in the environment to which the population was adapted and did not involve inbreeding. The results show an extraordinary similarity in the behavior of replicates of the same chromosome, indicating consistent genetic effects on total fitness. Some invading chromosomes increased rapidly and some slowly, and some rose to appreciable frequency after several months, but then declined again: in every case, the same pattern was seen in each replicate. We estimated relative fitnesses, rates of change of fitness, and relative viabilities, for each chromosome. There were significant fluctuations around the fitted model, which were also highly replicable. Wild-type chromosomes varied substantially in their effects on heterozygous fitness, and these effects vary through time, most likely as a result of genotype x environment interactions. PMID- 15545657 TI - Genetics of species differences in the wild annual sunflowers, Helianthus annuus and H. petiolaris. AB - Much of our knowledge of speciation genetics stems from quantitative trait locus (QTL) studies. However, interpretations of the size and distribution of QTL underlying species differences are complicated by differences in the way QTL magnitudes are estimated. Also, many studies fail to exploit information about QTL directions or to compare inter- and intraspecific QTL variation. Here, we comprehensively analyze an extensive QTL data set for an interspecific backcross between two wild annual sunflowers, Helianthus annuus and H. petiolaris, interpret different estimates of QTL magnitudes, identify trait groups that have diverged through selection, and compare inter- and intraspecific QTL magnitudes. Our results indicate that even minor QTL (in terms of backcross variance) may be surprisingly large compared to levels of standing variation in the parental species or phenotypic differences between them. Morphological traits, particularly flower morphology, were more strongly or consistently selected than life history or physiological traits. Also, intraspecific QTL were generally smaller than interspecific ones, consistent with the prediction that larger QTL are more likely to spread to fixation across a subdivided population. Our results inform the genetics of species differences in Helianthus and suggest an approach for the simultaneous mapping of inter- and intraspecific QTL. PMID- 15545658 TI - Estimation of long-term effective population sizes through the history of durum wheat using microsatellite data. AB - Estimation of long-term effective population size (N(e)) from polymorphism data alone requires an independent knowledge of mutation rate. Microsatellites provide the opportunity to estimate N(e) because their high mutation rate can be estimated from observed mutations. We used this property to estimate N(e) in allotetraploid wheat Triticum turgidum at four stages of its history since its domestication. We estimated the mutation rate of 30 microsatellite loci. Allele specific mutation rates mu were predicted from the number of repeats of the alleles. Effective population sizes were calculated from the diversity parameter theta = 4N(e)mu. We demonstrated from simulations that the unbiased estimator of theta based on Nei's heterozygosity is the most appropriate for estimating N(e) because of a small variance and a relative robustness to variations in the mutation model compared to other estimators. We found a N(e) of 32,500 individuals with a 95% confidence interval of [20,739; 45,991] in the wild ancestor of wheat, 12,000 ([5790; 19,300]) in the domesticated form, 6000 ([2831; 9556]) in landraces, and 1300 ([689; 2031]) in recent improved varieties. This decrease illustrates the successive bottlenecks in durum wheat. No selective effect was detected on our loci, despite a complete loss of polymorphism for two of them. PMID- 15545660 TI - Selection on Glycine beta-1,3-endoglucanase genes differentially inhibited by a Phytophthora glucanase inhibitor protein. AB - Plant endo-beta-1,3-glucanases (EGases) degrade the cell wall polysaccharides of attacking pathogens and release elicitors of additional plant defenses. Isozymes EGaseA and EGaseB of soybean differ in susceptibility to a glucanase inhibitor protein (GIP1) produced by Phytophthora sojae, a major soybean pathogen. EGaseA, the major elicitor-releasing isozyme, is a high-affinity ligand for GIP1, which completely inhibits it, whereas EGaseB is unaffected by GIP1. We tested for departures from neutral evolution on the basis of partial sequences of EGaseA and EGaseB from 20 widespread accessions of Glycine soja (the wild progenitor of soybean), from 4 other Glycine species, and across dicotyledonous plants. G. soja exhibited little intraspecific variation at either locus. Phylogeny-based codon evolution models detected strong evidence of positive selection on Glycine EGaseA and weaker evidence for selection on dicot EGases and Glycine EGaseB. Positively selected peptide sites were identified and located on a structural model of EGase bound to GIP1. Positively selected sites and highly variable sites were found disproportionately within 4.5 angstroms of bound GIP1. Low variation within G. soja EGases, coupled with positive selection in both Glycine and dicot lineages and the proximity of rapidly evolving sites to GIP1, suggests an arms race involving repeated adaptation to pathogen attack and inhibition. PMID- 15545659 TI - Many P-element insertions affect wing shape in Drosophila melanogaster. AB - A screen of random, autosomal, homozygous-viable P-element insertions in D. melanogaster found small effects on wing shape in 11 of 50 lines. The effects were due to single insertions and remained stable and significant for over 5 years, in repeated, high-resolution measurements. All 11 insertions were within or near protein-coding transcription units, none of which were previously known to affect wing shape. Many sites in the genome can affect wing shape. PMID- 15545661 TI - Effect of addition of adjuvant paclitaxel on radiotherapy delivery and locoregional control of node-positive breast cancer: cancer and leukemia group B 9344. AB - PURPOSE: We compared radiotherapy (RT) delivery and locoregional control in patients with node-positive breast cancer randomly assigned on Cancer and Leukemia Group B 9344 to receive adjuvant doxorubicin/cyclophosphamide (AC) with patients assigned to receive AC followed by paclitaxel (AC+T). METHODS: Eligible patients were randomly assigned to receive adjuvant AC versus AC+T chemotherapy. RT was required if breast-conserving surgery was performed but was elective after mastectomy. Information about RT delivery was retrospectively collected. Cumulative incidence of locoregional recurrence (LRR), use of elective RT, and RT delivery were compared between treatment arms. RESULTS: For patients treated with breast-conserving surgery and RT, the 5-year cumulative incidence of isolated LRR was 9.7% in the AC arm and 3.7% in the AC+T arm (P = .04) and of LRR as any component of failure was 12.9% versus 6.1%, respectively (P = .04). Although LRR rates in patients who did not receive postmastectomy RT were lower in the AC+T arm, the difference was not statistically significant. Despite the lack of protocol guidelines, RT use did not differ between arms, nor did RT dose, treatment interruption, or completion. CONCLUSION: Despite the delay to RT during additional chemotherapy, adjuvant AC+T afforded better local control than AC alone in patients treated with breast-conserving therapy. Addition of paclitaxel did not adversely affect delivery or ability to tolerate RT, as indicated by similar rates of completion of timely, full-dose RT between arms. PMID- 15545662 TI - Chemotherapy and radiation therapy for breast cancer: what is the optimal sequence? PMID- 15545663 TI - Targeted therapy with imatinib: hits and misses? PMID- 15545664 TI - American Society of Clinical Oncology technology assessment on the use of aromatase inhibitors as adjuvant therapy for postmenopausal women with hormone receptor-positive breast cancer: status report 2004. AB - PURPOSE: To update the 2003 American Society of Clinical Oncology technology assessment on adjuvant use of aromatase inhibitors. RECOMMENDATIONS: Based on results from multiple large randomized trials, adjuvant therapy for postmenopausal women with hormone receptor-positive breast cancer should include an aromatase inhibitor in order to lower the risk of tumor recurrence. Neither the optimal timing nor duration of aromatase inhibitor therapy is established. Aromatase inhibitors are appropriate as initial treatment for women with contraindications to tamoxifen. For all other postmenopausal women, treatment options include 5 years of aromatase inhibitors treatment or sequential therapy consisting of tamoxifen (for either 2 to 3 years or 5 years) followed by aromatase inhibitors for 2 to 3, or 5 years. Patients intolerant of aromatase inhibitors should receive tamoxifen. There are no data on the use of tamoxifen after an aromatase inhibitor in the adjuvant setting. Women with hormone receptor negative tumors should not receive adjuvant endocrine therapy. The role of other biomarkers such as progesterone receptor and HER2 status in selecting optimal endocrine therapy remains controversial. Aromatase inhibitors are contraindicated in premenopausal women; there are limited data concerning their role in women with treatment-related amenorrhea. The side effect profiles of tamoxifen and aromatase inhibitors differ. The late consequences of aromatase inhibitor therapy, including osteoporosis, are not well characterized. CONCLUSION: The Panel believes that optimal adjuvant hormonal therapy for a postmenopausal woman with receptor-positive breast cancer includes an aromatase inhibitor as initial therapy or after treatment with tamoxifen. Women with breast cancer and their physicians must weigh the risks and benefits of all therapeutic options. PMID- 15545665 TI - Impact of concurrent versus sequential tamoxifen with radiation therapy in early stage breast cancer patients undergoing breast conservation treatment. AB - PURPOSE: To assess the impact of sequencing of tamoxifen and radiation therapy (RT) on outcomes in early-stage breast cancer. PATIENTS AND METHODS: This retrospective study evaluates the effect of the sequence of tamoxifen with RT on outcomes in stage I to II breast cancer patients who underwent breast conservation treatment (BCT) and received adjuvant tamoxifen, with or without adjuvant chemotherapy. Patients were grouped as concurrent (tamoxifen given during RT followed by continued tamoxifen; 174 patients) and sequential (RT followed by tamoxifen; 104 patients). RESULTS: Median follow-up after RT was 8.6 years for both groups. The pathologic T and N stage, race, estrogen and progesterone status, number of positive nodes, and RT were comparable between the two groups (all P >/= .08). More women age 49 years or younger and women who received chemotherapy were in the sequential group than the concurrent group (6% and 25%, respectively; P < .0001). The sequence of tamoxifen therapy did not influence 10-year local recurrence rates (sequential, 7%; concurrent, 3%; P = .52), overall survival (sequential, 86%; concurrent, 81%; P = .64), or relapse free survival (sequential, 76%; concurrent, 85%; P = .35). When adjusting age and chemotherapy use in the multivariable Cox model, hazard ratios comparing sequential versus concurrent tamoxifen therapy were 1.56 (95% CI, 0.87 to 2.79), 1.23 (95% CI, 0.63 to 2.41), and 1.22 (95% CI, 0.33 to 4.49) for the overall survival, relapse-free survival, and local recurrence, respectively. CONCLUSION: The therapeutic regimens of tamoxifen given concurrently or sequentially with RT both appear to be reasonable options for patients treated with BCT. PMID- 15545666 TI - Sequence of radiotherapy with tamoxifen in conservatively managed breast cancer does not affect local relapse rates. AB - PURPOSE: To evaluate whether the sequencing of tamoxifen (TAM) relative to radiation (RT) affects outcome in breast cancer patients treated with conservative surgery (CS) plus RT (lumpectomy with RT). METHODS: Between 1976 and 1999, 1,649 patients with stage I or II breast cancer were treated with CS plus RT at Yale-New Haven Hospital (New Haven, CT). TAM was administered to 500 patients. The timing of TAM relative to RT was documented for each patient. Of the 500 patients, the timing of TAM was unclear in five patients, was administered concurrently with RT in 254 patients (CON-TAM), and was administered sequentially after completion of RT in 241 patients (SEQ-TAM). RESULTS: There were no differences between the CON-TAM and SEQ-TAM group in T stage, estrogen and progesterone status, nodal status, histology, or margin status. The CON-TAM group was slightly older than the SEQ-TAM group (62 v 58 years) and received chemotherapy in addition to TAM less frequently (14% v 38%). As of September 2002, with a median follow-up of 10.0 years, there were no significant differences between the CON-TAM and SEQ-TAM groups in overall survival (84% v 82%; hazard ratio [HR], 1.234; 95% CI, 0.42 to 2.05; P = .45), distant-metastasis free rate (82% v 78%; HR, 1.55; 95% CI, 0.89 to 2.68; P = .12), ipsilateral breast-relapse-free rate (90% v 86%; HR, 0.932; 95% CI, 0.42 to 2.05; P = .86), or contralateral breast-relapse-free rate (95% v 93%; HR, 0.892; 95% CI, 0.53 to 1.48; P = .66). CONCLUSION: Although the concurrent use of TAM with RT may theoretically render cancer cells less responsive to RT, this retrospective study suggests that in practical application, concurrent administration of TAM with RT does not compromise local control. PMID- 15545667 TI - Radiation therapy and tamoxifen: concurrent or sequential? That is the question. PMID- 15545668 TI - KIT and platelet-derived growth factor receptor alpha tyrosine kinase gene mutations and KIT amplifications in human solid tumors. AB - PURPOSE: Mutated KIT and platelet-derived growth factor receptor alpha (PDGFRalpha) tyrosine kinases are the principal targets for imatinib mesylate in the treatment of gastrointestinal stromal tumors (GISTs). The frequency of activating KIT and PDGFRA gene mutations in most other histologic types of human cancer is not known. MATERIALS AND METHODS: KIT exons 9, 11, 13, and 17 and PDGFRA exons 11 and 17 of 334 human cancers were screened for mutations using sensitive denaturing high-performance liquid chromatography (DHPLC). In addition, all KIT exons from 9 to 21 of 115 tumors were screened. Thirty-two histologic tumor types were examined. Samples with abnormal findings in DHLPC were sequenced. Immunostaining for the KIT protein (CD117) was performed in 322 (96.4%) of the 334 cases. RESULTS: Of the 3,039 exons screened, only 17 had mutation. All 17 cases with either mutated KIT (n = 15) or PDGFRA (n = 2) were histologically GIST tumors, whereas none of the other histologic types of cancer (n = 316) harbored KIT or PDGFRA mutation. KIT immunostaining was rarely positive except in GISTs (18 of 18), small-cell lung cancer (10 of 30; 33%), and testicular teratocarcinoma (four of 17; 24%). Wild-type KIT gene amplification or chromosome 4 aneuploidy was common (seven of 12) in non-GIST tumors with strong KIT protein expression when studied with fluorescence in situ hybridization. CONCLUSION: Despite frequent KIT protein expression in some tumor types, KIT and PDGFRA gene mutations are uncommon in most human cancers. Cancer KIT expression is frequently associated with multiple copies of the wild-type KIT gene. PMID- 15545670 TI - Summaries for patients. What is the appropriate level of blood-thinning medication for elderly people with atrial fibrillation? PMID- 15545669 TI - Sequencing of tamoxifen and radiotherapy after breast-conserving surgery in early stage breast cancer. AB - PURPOSE: Tamoxifen (TAM) is thought to exert a cytostatic effect on hormone sensitive breast cancer cells. Some preclinical studies show reduced radiosensitivity in irradiated malignant mammary epithelial cells when pretreated with TAM; other studies refute these results. Recent randomized clinical trials suggest an antagonistic effect of TAM on cytotoxic therapy, with improved disease free survival (DFS) with sequential versus concurrent TAM. An exploratory analysis was undertaken to evaluate the optimal sequencing of TAM and radiotherapy (RT) after breast-conserving surgery. PATIENTS AND METHODS: Southwest Oncology Group trial 8897 (Intergroup 0102) randomly assigned node negative women with T1-3 breast cancers to cyclophosphamide, doxorubicin, fluorouracil (CAF); CAF --> TAM; cyclophosphamide, methotrexate, fluorouracil (CMF); and CMF --> TAM. For this analysis, data are reported only in the TAM groups. RT was allowed either before adjuvant therapy (sequential [SEQ] RT; 107 patients) or after chemotherapy but concurrent with TAM (concurrent [CONC] RT; 202 patients). Survival data were adjusted for receptor status, age, and tumor size. RESULTS: With a median follow-up of 10.3 years, 10-year DFS values were 83% and 83% for CONC versus SEQ RT groups (log-rank P = .73; P = .76 adjusted for patient characteristics), and 10-year overall survivals were 88% and 90%, respectively (log-rank P = .59; adjusted P = .65). Patterns of failure showed no increase in in-breast recurrence rates between CONC RT and SEQ RT groups, with 10 year local recurrence rates of 7% for CONC RT and 5% for SEQ RT (hazard ratio, 0.73; 95% CI, 0.26 to 2.04; P = .54). CONCLUSION: The current analysis does not suggest an adverse effect on local or systemic control with CONC versus SEQ TAM and RT in node-negative breast cancer. A randomized trial is encouraged to validate these results. PMID- 15545671 TI - Summaries for patients. Detecting pancreatic cancer and its spread. PMID- 15545672 TI - Summaries for patients. Glucocorticoid medications and the risk for cardiovascular disease. PMID- 15545673 TI - Summaries for patients. The use of clinical vignettes to measure the quality of health care. PMID- 15545674 TI - Advanced age, anticoagulation intensity, and risk for intracranial hemorrhage among patients taking warfarin for atrial fibrillation. AB - BACKGROUND: The risk for atrial fibrillation-associated stroke increases at low anticoagulation intensities. However, higher intensities increase hemorrhage risk. Optimal use of warfarin for atrial fibrillation requires precise information on the risk for intracranial hemorrhage as a function of patient age and anticoagulation intensity. OBJECTIVE: To examine the relationship of age, anticoagulation intensity, and risk for intracranial hemorrhage. DESIGN: Case control study. SETTING: Academic medical center. PATIENTS: 170 case-patients who developed intracranial hemorrhage during warfarin therapy and 1020 matched controls who did not; both case-patients and controls were taking warfarin for atrial fibrillation. MEASUREMENTS: The authors performed multivariable conditional logistic regression to determine the odds of intracranial hemorrhage with regard to age and international normalized ratio (INR), controlling for comorbid conditions and aspirin use. RESULTS: Case-patients were older than controls (median age, 78 years vs. 75 years; P < 0.001) and had higher median INRs (2.7 vs. 2.3; P < 0.001). The risk for intracranial hemorrhage increased at 85 years of age or older (adjusted odds ratio, 2.5 [95% CI, 1.3 to 4.7]; referent age, 70 to 74 years) and at an INR range of 3.5 to 3.9 (adjusted odds ratio, 4.6 [CI, 2.3 to 9.4]; referent INR, 2.0 to 3.0). The risk for intracranial hemorrhage at INRs less than 2.0 did not differ statistically from the risk at INRs of 2.0 to 3.0 (adjusted odds ratio, 1.3 [CI, 0.8 to 2.2]). LIMITATIONS: Although duration of anticoagulation has been associated with hemorrhage in other studies, the current study could not control for this potential confounder. CONCLUSIONS: The risk for intracranial hemorrhage increases at age 85 years. International normalized ratios less than 2.0 were not associated with lower risk for intracranial hemorrhage compared with INRs between 2.0 and 3.0. Therefore, anticoagulation management should focus on maintaining INRs in the 2.0 to 3.0 range, even in elderly patients with atrial fibrillation, rather than targeting INRs less than 2.0. Similarly, INRs of 3.5 or greater should be avoided. PMID- 15545675 TI - Comparison of endoscopic ultrasonography and multidetector computed tomography for detecting and staging pancreatic cancer. AB - BACKGROUND: Accurate preoperative detection and staging of pancreatic cancer may identify patients with locoregional disease that is amenable to surgical resection. OBJECTIVE: To compare endoscopic ultrasonography and multidetector computed tomography (CT) for the detection, staging, and resectability of known or suspected locoregional pancreatic cancer. DESIGN: Prospective, observational, cohort study. SETTING: Single, tertiary referral hospital in Indianapolis, Indiana. PATIENTS: 120 participants with known or suspected locoregional pancreatic cancer. INTERVENTIONS: Endoscopic ultrasonography followed by multidetector CT was performed in all patients. Patients with known or suspected pancreatic cancer deemed potentially resectable by 1 or both tests were considered for surgery. MEASUREMENTS: Detection, staging, and resectability of pancreatic cancer. Surgically resected pancreatic cancer with negative microscopic histologic margins was considered resectable. RESULTS: Of 120 patients enrolled, 104 (87%) underwent endoscopic ultrasonography and CT. Of the 80 patients with pancreatic cancer, 27 (34%) were managed nonoperatively, and 53 (66%) treated surgically had resectable (n = 25) or unresectable (n = 28) cancer. For the 80 patients with cancer, the sensitivity of endoscopic ultrasonography (98% [95% CI, 91% to 100%]) for detecting a pancreatic mass was greater than that of CT (86% [CI, 77% to 93%]; P = 0.012). For the 53 surgical patients, endoscopic ultrasonography was superior to CT for tumor staging accuracy (67% vs. 41%; P < 0.001) but equivalent for nodal staging accuracy (44% vs. 47%; P > 0.2). Of the 25 resectable pancreatic tumors in patients recommended for surgery, endoscopic ultrasonography and CT correctly identified 88% and 92%, respectively, as resectable. Of the 28 unresectable pancreatic tumors in patients recommended for surgery, endoscopic ultrasonography and CT correctly identified 68% and 64%, respectively, as unresectable. LIMITATIONS: Radiologists who read the scans and endosonographers were not blinded to previous radiographic information. Because of the modest sample size, CIs of the sensitivity estimates were sometimes wide. CONCLUSION: Compared with multidetector CT, endoscopic ultrasonography is superior for tumor detection and staging but similar for nodal staging and resectability of preoperatively suspected nonmetastatic pancreatic cancer. PMID- 15545676 TI - Taking glucocorticoids by prescription is associated with subsequent cardiovascular disease. AB - BACKGROUND: Glucocorticoids have adverse systemic effects, including obesity, hypertension, and hyperglycemia, that may predispose to cardiovascular disease. The effect of glucocorticoid use on cardiovascular disease has not been quantified. OBJECTIVE: To test the hypothesis that users of exogenous glucocorticoids have an increased risk for cardiovascular disease. DESIGN: A cohort study using a record linkage database. SETTING: Tayside, Scotland, United Kingdom. PATIENTS: 68,781 glucocorticoid users and 82,202 nonusers without previous hospitalization for cardiovascular disease who were studied between 1993 and 1996. MEASUREMENTS: The average daily dose of glucocorticoid exposure during follow-up was categorized as low (inhaled, nasal, and topical only), medium (oral, rectal, or parenteral <7.5 mg of prednisolone equivalent), or high (> or =7.5 mg of prednisolone equivalent). Poisson regression model, sensitivity analysis, and propensity score methods were used to investigate the association between glucocorticoid exposure and cardiovascular outcome. RESULTS: 4383 cardiovascular events occurred in 257,487 person-years of follow-up for a rate of 17.0 (95% CI, 16.5 to 17.5) per 1000 person-years in the comparator group, and 5068 events occurred in 212,287 person-years for a rate of 23.9 (CI, 23.2 to 24.5) per 1000 person-years in the group exposed to glucocorticoids (22.1, 27.2, and 76.5 in low, medium, and high groups, respectively). The absolute risk difference was 6.9 (CI, 6.0 to 7.7) per 1000 person-years (5.1, 10.1, and 59.4, respectively). After adjustment for known covariates, the relative risk for a cardiovascular event in patients receiving high-dose glucocorticoids was 2.56 (CI, 2.18 to 2.99). LIMITATIONS: Because the data were observational, residual confounding cannot be excluded. CONCLUSION: Treatment with high-dose glucocorticoids seemed to be associated with increased risk for cardiovascular disease. PMID- 15545677 TI - Measuring the quality of physician practice by using clinical vignettes: a prospective validation study. AB - BACKGROUND: Worldwide efforts are under way to improve the quality of clinical practice. Most quality measurements, however, are poorly validated, expensive, and difficult to compare among sites. OBJECTIVE: To validate whether vignettes accurately measure the quality of clinical practice by using a comparison with standardized patients (the gold standard method), and to determine whether vignettes are a more or less accurate method than medical record abstraction. DESIGN: Prospective, multisite study. SETTING: Outpatient primary care clinics in 2 Veterans Affairs medical centers and 2 large, private medical centers. PARTICIPANTS: 144 of 163 eligible physicians agreed to participate, and, of these, 116 were randomly selected to see standardized patients, to complete vignettes, or both. MEASUREMENTS: Scores, expressed as the percentage of explicit quality criteria correctly completed, were obtained by using 3 methods. RESULTS: Among all physicians, the quality of clinical practice as measured by the standardized patients was 73% correct (95% CI, 72.1% to 73.4%). By using exactly the same criteria, physicians scored 68% (CI, 67.9% to 68.9%) when measured by the vignettes but only 63% (CI, 62.7% to 64.0%) when assessed by medical record abstraction. These findings were consistent across all diseases and were independent of case complexity or physician training level. Vignettes also accurately measured unnecessary care. Finally, vignettes seem to capture the range in the quality of clinical practice among physicians within a site. LIMITATIONS: Despite finding variation in the quality of clinical practice, we did not determine whether poorer quality translated into worse health status for patients. In addition, the quality scores are based on measurements from 1 patient-provider interaction. As with all other scoring criteria, vignette criteria must be regularly updated. CONCLUSIONS: Vignettes are a valid tool for measuring the quality of clinical practice. They can be used for diverse clinical settings, diseases, physician types, and situations in which case-mix variation is a concern. They are inexpensive and easy to use. Vignettes are particularly useful for comparing quality among and within sites and may be useful for longitudinal evaluations of interventions intended to change clinical practice. PMID- 15545678 TI - Better reporting of harms in randomized trials: an extension of the CONSORT statement. AB - In response to overwhelming evidence and the consequences of poor-quality reporting of randomized, controlled trials (RCTs), many medical journals and editorial groups have now endorsed the CONSORT (Consolidated Standards of Reporting Trials) statement, a 22-item checklist and flow diagram. Because CONSORT primarily aimed at improving the quality of reporting of efficacy, only 1 checklist item specifically addressed the reporting of safety. Considerable evidence suggests that reporting of harms-related data from RCTs also needs improvement. Members of the CONSORT Group, including journal editors and scientists, met in Montebello, Quebec, Canada, in May 2003 to address this problem. The result is the following document: the standard CONSORT checklist with 10 new recommendations about reporting harms-related issues, accompanying explanation, and examples to highlight specific aspects of proper reporting. We hope that this document, in conjunction with other CONSORT-related materials (http://www.consort-statement.org), will help authors improve their reporting of harms-related data from RCTs. Better reporting will help readers critically appraise and interpret trial results. Journals can support this goal by revising Instructions to Authors so that they refer authors to this document. PMID- 15545679 TI - The physiologic basis of high-altitude diseases. PMID- 15545680 TI - Update in rheumatology. PMID- 15545681 TI - Update on the health disparities literature. PMID- 15545682 TI - Back to the future: clinical vignettes and the measurement of physician performance. PMID- 15545683 TI - Inequality in health care: unjust, inhumane, and unattended! PMID- 15545684 TI - Medicine in the family. PMID- 15545685 TI - Improving improvement. PMID- 15545686 TI - Helicobacter pylori and idiopathic thrombocytopenic purpura. PMID- 15545687 TI - Improving improvement. PMID- 15545688 TI - Translating "tight control"? PMID- 15545689 TI - Acute hepatotoxicity caused by repaglinide. PMID- 15545690 TI - The health care revival in Iraq. PMID- 15545691 TI - Healthcare reform: responding to the rhetoric. PMID- 15545692 TI - Considering patient priorities when choosing a dressing. PMID- 15545693 TI - What ostomy patients still don't know 54 years later. PMID- 15545694 TI - Fall brings more changes. PMID- 15545695 TI - An innovative teaching strategy for pressure ulcer documentation: the case for computer-based learning. PMID- 15545696 TI - A three year multiphase pressure ulcer prevalence/incidence study in a regional referral hospital. AB - Pressure ulcers can have a devastating impact on health and care provision, ranging from patient discomfort and increased healthcare costs to a potential reflection on the quality of care. To evaluate the outcomes of prevention education and skin integrity interventions on the incidence of pressure ulcers, a multiphase project was initiated in an urban 154-bed regional referral community hospital in Ontario, Canada that provides care to an urban and rural population. The prevalence study included 84 adult subjects at baseline, 77 after one year (Phase 1), and 100 after 3 years (Phase 2). The Braden Scale for Predicting Pressure Sore Risk and a data collection form were used to record prevalence, incidence, stage and location of pressure ulcers, and related documented interventions. Incidence data were obtained from patient charts and defined as ulcers that developed over 24 hours following admission. Phase 1 interventions involved staff education and replacement of existing skin care products. Phase 2 interventions included adoption of pressure ulcer prevention protocols, advanced wound care products, improved support surface usage, modification of documentation methods, and staff education. Of the 84 patients assessed at baseline , 15 (17.9 %) developed 22 pressure ulcers compared to 4 of 77 (5.2%) during Phase 1 and 2 out of 100 (2.0%) during Phase 2. The difference between baseline and both subsequent time points was statistically significant (P greater than 0.05). These results suggest that education and the implementation of appropriate skin care products and procedures and pressure ulcer prevention protocols may reduce the incidence of hospital-acquired pressure ulcers. PMID- 15545697 TI - Comparing a foam composite to a hydrocellular foam dressing in the management of venous leg ulcers: a controlled clinical study. AB - Venous leg ulcers are the most prevalent form of chronic wounds in the Western world. The principles of moist wound healing coupled with the use of graduated compression bandaging have become the cornerstone of treatment for venous leg ulcers but not all moist dressings are alike. To compare the attributes of a foam composite dressing with those of a hydrocellular foam dressing in the management of venous leg ulcers, a prospective, randomized, comparative 12-week study was conducted in 15 centers in the US, Canada, France, Germany, and the UK. Dressings were changed and compression bandages applied per manufacturer recommendations and dressing performance was assessed at every dressing change and at the final evaluation. Patients with venous ulcers were randomized to treatment for 12 weeks with either hydrocellular foam (n = 52) or foam composite (n = 55) dressing. Healing differences between the groups were not statistically significant, with 36% of patients using foam composite dressing healed in a mean of 66 days and 39% of patients using hydrocellular foam dressing healed in a mean of 73 days. However, the foam composite dressing performed significantly better than the hydrocellular foam dressing with regard to condition of the periwound skin - 55% of patients in the foam composite group having healed or markedly improved surrounding skin compared to 37% of patients using hydrocellular foam (P = 0.03). The foam composite dressing was rated significantly better than the hydrocellular foam dressing ("very good" to "excellent") in level of satisfaction with conformability (87% and 75%, respectively, P = 0.05); being non-sensitizing (73% and 52%, respectively, P = 0.02); and ease of application (93% and 81%, respectively, P = 0.01). The findings reported in this study suggest that the foam composite dressing offers significant improvements in the quality of life of patients with venous leg ulcers as well as for their caregivers. PMID- 15545698 TI - Wound instillation--the next step in negative pressure wound therapy. Lessons learned from initial experiences. AB - Negative pressure wound therapy uses a reticulated sponge and subatmospheric pressure to facilitate healing of a variety of wounds. The therapy appears to assist wound healing by decreasing wound bacterial burden and edema while facilitating granulation tissue formation. The latest development in negative pressure wound therapy allows clinicians to instill a solution into the wound and is indicated for use when patients receiving negative pressure wound therapy would benefit from the controlled delivery of topical wound treatment solutions and suspensions. A retrospective analysis of five cases was conducted to describe initial experiences with instillation and negative pressure wound therapy (average treatment time on negative pressure wound therapy with instillation 15 days, range 5 to 24 days). In two of the five cases where traditional negative pressure wound therapy was tried, infected wounds improved and culture results became negative following instillation of antibiotics in a saline solution. In all cases where used, instillation of a topical anesthetic appeared to effectively minimize wound pain that is occasionally associated with negative pressure wound therapy. Negative pressure wound therapy with instillation and possibly hyperbaric oxygen therapy may be initiated from the onset of treatment to manage high-risk wounds that appear severely infected and may be limb threatening if located on an extremity. Controlled clinical studies to ascertain the efficacy and cost-effectiveness of this treatment modality are needed. PMID- 15545699 TI - Erythrocyte osmotic fragility and oxidative stress in experimental hypothyroidism. AB - The present study was planned to explain the relation between erythrocyte osmotic fragility and oxidative stress and antioxidant statue in primary hypothyroid induced experimental rats. Twenty-four Spraque Dawley type female rats were divided into two, as control (n = 12) and experimental (n = 12), groups weighing between 160 and 200 g. The experimental group animals have received tap water methimazole added standard fodder to block the iodine pumps for 30 d (75 mg/100 g). Control group animals were fed tap water and only standard fodder for the same period. At the end of 30 d blood samples were drawn from the abdominal aorta of the rats under ether anesthesia. T3, T4, and TSH levels were measured and the animals that had relatively lower T3, T4, and higher TSH levels were accepted as hypothyroid group. Hormone levels of the control group were at euthyroid conditions. Osmotic fragility, as a lipid peroxidation indicator malondialdehyde (MDA), antioxidant defense system indicators superoxide dismutase (SOD) and glutathione (GSH) levels were measured in the blood samples. Osmotic fragility test results: There was no statistically significant difference found between maximum osmotic hemolysis limit values of both group. Minimum osmotic hemolysis limit value of hypothyroid group was found to be higher than that of control group values (p < 0.02). The standard hemolysis and hemolytic increment curve of the hypothyroid group drawn according to osmotic fragility test results was found to be shifted to the right when compared to control group's curve. This situation and hemolytic increment value, which shows maximum hemolysis ratio, is the proof of increased osmotic fragility of the erythrocytes in hypothyroidism. There is no statistically significant difference found between hypothyroid and control groups in the lipid peroxidation indicator MDA and antioxidant indicators SOD and GSH levels. As a result of our study it may be concluded that hypothyroidism may lead to an increase in osmotic fragility of erythrocytes. But the increase in erythrocyte osmotic fragility does not originate from lipid peroxidation. PMID- 15545700 TI - Plasma concentrations of ir-inhibin, inhibin A, inhibin pro-alphaC, FSH, and estradiol-17beta during estrous cycle in mares and their relationship with follicular growth. AB - The relationship among plasma levels of immunoreactive (ir)-inhibin, inhibin A, inhibin pro-alphaC, FSH, estradiol-17beta and follicular growth were investigated during the normal estrous cycle in mares. Seven mares were used for two successive normal estrous cycles. Follicular diameters and ovulation were obtained by transrectal ultrasonography, and blood samples were collected daily by jugular venipuncture for hormonal assay. The results showed that inhibin A was inversely correlated (r = -0.59, p < 0.0001) with FSH indicating its contribution to negative feedback control of FSH secretions from the pituitary gland. Estradiol-17beta increased during the follicular phase reaching a peak (37.9 +/- 3.8 pg/mL) 2 d before ovulation. Estradiol-17beta was positively correlated (r = 0.78, p < 0.0001) with inhibin A. The high levels of inhibin A and estradiol 17beta were associated with the growth of the preovulatory dominant follicle and inversely correlated with FSH suggesting that both hormones are products of the large dominant follicles and were responsible for the decline in FSH secretion during the follicular phase of estrous cycle. In conclusion, an inverse relationship between inhibin A and FSH was clearly demonstrated indicating that inhibin A has a key role in the negative feedback control of FSH from the pituitary gland. In addition, inhibin A and estra-diol-17beta secretions were associated with the growth of the preovulatory dominant follicle and were positively correlated. PMID- 15545701 TI - Age-related decreases in gonadal hormones in Long-Evans rats: relationship to rise in arterial pressure. AB - Sex steroids modify sexual behavior and autonomic function. The gradual decline in circulating levels is correlated with several diseases in humans and animals. However, little is known about age-related changes that occur in the availability of these steroids. In the current studies, we characterized age-related changes in (1) circulating levels of estradiol (females) or testosterone (males), (2) reproductive function (estrous cyclicity in females; erectile reflexes in males), and (3) blood pressure in a longitudinal study. In a separate study, we characterized the estrous cyclicity of sex steroids in female, and diurnal periodicity in male, Long-Evans rats. Young females exhibit regular estrous cycles, transition to irregular cycles at about 10 mo of age, then to cycles characterized by extended periods of estrous, and to persistent estrous. Despite the loss of cyclicity, circulating 17beta-estradiol in middle-aged females was maintained at levels similar to those in young females during diestrous. Males display an age-related decline in testosterone, circulating levels decrease by about 25% during the period from 8 to 16 mo of age. Also, during any 24 h period testosterone levels in young males vary from a peak of about 3.5 ng/mL (late light period) to a trough of 0.7 ng/mL (early dark period). In middle-aged males the rhythm amplitude is greatly blunted (1.4 to 0.7 ng/mL). Males exhibit age related decrements in erectile reflexes. In females and males systolic blood pressure is relatively stable until 8 mo of age, but significantly increases during the next 5 mo of age. In males, the increase in arterial pressure is gradual from about 8 mo of age. Young females have lower blood pressures than age matched males, but by 14 mo of age this sex-related advantage is lost. Thus, by middle age, male and female rats are exposed to less gonadal hormone/altered patterns of availability, exhibit decrements in reproductive function, and display an increase in systolic blood pressure. PMID- 15545702 TI - Stimulation by D-glucose of 36Cl- efflux from prelabeled rat pancreatic islets. AB - D-glucose was previously reported to cause a concentration-related decrease in the 36Cl- content of prelabeled islets prepared from ob/ob mice, a current animal model of inherited obesity. From these findings, it was inferred that the hexose stimulates Cl- efflux from islet cells and that such an increase in Cl- permeability may partly mediate glucose-induced depolarization of insulin producing cells. The aim of the present study was to investigate the possible extension of these findings to islets prepared from normal rats by measuring the changes evoked by increasing concentrations of D-glucose in 36Cl- outflow itself from prelabeled isolated islets. After 60 min preincubation at 37 degrees C in the presence of 3 mM D-glucose and 36Cl- (75 microCi/mL), the islets were incubated for 8-10 min at 37 degrees C in the presence of increasing concentrations of the hexose (3-20 mM). The changes in 36Cl- outflow during incubation indicated that D-glucose, in excess of a threshold concentration close to 5 mM, indeed increases effluent radioactivity from the prelabeled islets. It is proposed, therefore, that a gating of volume-sensitive anion channels in glucose-stimulated insulin-producing islet cells participates in the depolarization of the plasma membrane recorded in the range of insulinotropic concentrations of the hexose. PMID- 15545703 TI - Androgen receptor regulates expression of skeletal muscle-specific proteins and muscle cell types. AB - C2C12 myoblasts expressing the androgen receptor (AR) were used to analyze the role of androgen-AR signaling pathway in skeletal muscle development. Marked up regulation of AR expression was observed in differentiated myotubes. A nuclear run-on transcription assay demonstrated that transcription of the AR gene is increased during skeletal muscle cell differentiation. Regulation of skeletal muscle-specific protein expression by the androgen-AR signaling pathway was further analyzed using quadriceps skeletal muscle from wild-type (WT) and AR knock-out (ARKO) male mice. A histological analysis of quadriceps skeletal muscle indicates no morphological differences between ARKO and WT mice. However, the androgen-AR signaling pathway increases expression of slow-twitch-specific skeletal muscle proteins and downregulates fast-twitch-specific skeletal muscle proteins, resulting in an increase of slow-twitch muscle fiber type cells in quadriceps muscle. PMID- 15545704 TI - Effect of recombinant human growth hormone on age-related hepatocyte changes in old male and female Wistar rats. AB - Aging induces changes in several organs, such as the liver, and this process might be due to damage caused by free radicals and inflammatory mediators. The growth hormone (GH)/insulin-like growth factor-1 (IGF-1) axis shows a reduction with age, and this fact could be associated with some age-related changes. The aim of this study was to investigate the effect of GH administration on age induced alterations in hepatocytes. Two and twenty two month-old male and female Wistar rats were used. Old rats were treated with human recombinant GH for 10 wk. At the end of the treatment, hepatocytes were isolated from the liver and cultured, and different parameters were measured in cells and medium. Plasma IGF 1 was also measured. Aging significantly decreased plasma IGF-1 in males. In females, plasma IGF-1 was also reduced, but not significantly. GH treatment restored plasma IGF-1 levels to values similar to young males. Aging was associated with a significant increase in lipid peroxidation (LPO), nitric oxide (NO), carbon monoxide (CO) and cyclic guanosyl-monophosphate (cGMP), as well as a reduction in adenosyl triphosphate (ATP) and phosphatidylcholine (PC) synthesis. GH administration partially prevented all these changes in males. In females, some of the parameters were significantly improved by GH (ATP, CO, cGMP), while others showed a tendency to improvement, although differences did not reach significance. In conclusion, GH administration could exert beneficial effects against age-related changes in hepatocytes, mainly in males. PMID- 15545705 TI - 24-hour pattern of circulating prolactin and growth hormone levels and submaxillary lymph node immune responses in growing male rats subjected to social isolation. AB - To assess the effect of social isolation of growing rats on 24-h rhythmicity of circulating prolactin and growth hormone (GH) levels and submaxillary lymph node immune responses, male Wistar rats were either individually caged or kept in groups (4-5 animals per cage) for 30 d starting on d 35 of life. Plasma prolactin and GH levels, and submaxillary lymph node lymphocyte subset populations, interferon (IFN)-gamma release and mitogenic responses to concanavalin A (Con A) and lipopolysaccharide (LPS) were determined at six time intervals during the 24 h span. Social isolation brought about changes in mean values and 24-h pattern of plasma prolactin and GH levels and lymph node immune responses. After isolation, prolactin and GH mean values decreased, and lymph node T, B, non T-non B, CD8+, and CD4+-CD8+ cells augmented, whereas lymph node CD4+/CD8+ ratio, IFN-gamma release and mitogenic responses decreased. Social isolation resulted in disruption of 24 h rhythmicity of every immune parameter tested. CD4+/CD8+ ratio, IFN-gamma release and Concanavalin A (Con A) and lipopolysaccharide (LPS) responses correlated significantly with plasma prolactin or GH levels while T/B ratio correlated with plasma prolactin levels only. B, non T-non B, and CD4+-CD8+ cells correlated negatively with plasma prolactin. Modifications in mean value and 24-h rhythmicity of plasma prolactin and GH levels are presumably involved in the effect of social isolation on immune responsiveness. PMID- 15545706 TI - Regulation of follicle-stimulating and luteinizing hormone receptor signaling by. AB - Follicle-stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) belong to the super-family of G protein-coupled receptors (GPCR); GPCRs are negatively regulated by RGS ("regulators of G protein signaling") proteins. In this study we evaluated the effects of RGS3 and RGS10 on FSHR and LHR ligand binding and effector coupling. FSHR and LHR ligand binding were unchanged in the presence of RGS3 or RGS10. However, signaling by FSHR and LHR was diminished by RGS3 but not by RGS10. This constitutes the first demonstration of an interaction between RGS proteins and LH and FSH signaling pathways and identifies a mechanism for negative regulation of RGS3 on FSHR and LHR signaling. PMID- 15545707 TI - Radioiodine treatment of hyperthyroidism: prognostic factors affecting outcome. AB - OBJECTIVE: To assess the effectiveness of radioactive iodine (RAI) treatment in patients with hyperthyroidism and to evaluate prognostic factors affecting outcome. RESEARCH DESIGN AND METHODS: Our cohort comprised 115 consecutive patients with hyperthyroidism treated with RAI at the Endocrinology Clinic at the Farabi Hospital, Trabzon, between 1994 and 2002. Data were retrieved from the endocrinology clinic database. Patients were categorized into three diagnostic groups: Graves' disease (GD), toxic multinodular (TMN) hyperthyroidism, and toxic adenoma. Our policy, over the period of the study, was to offer a single fixed first dose (10 mCi) 131I to all patients with toxic nodular goiter (TNG) for the first time and to all patients with relapsed GD. RESULTS: There was no significant difference in the cure rate between GD and TNG, but Graves' patients had a significantly higher incidence of hypothyroidism (p < 0.001). In contrast, incidence of euthyroidism was significantly increased in TNG than those of the patients with GD (p < 0.05). The incidences of hyperthyroidism, euthyroidism, cure rate, and persistent hyperthyroidism did not vary significantly between females and males. Age at onset of hyperthyroidism at diagnosis was not associated with outcome of RAI therapy. The incidence of hypothyroidism in patients who had nonpalpable goiter was higher than those in patients who had medium or large goiter (p < 0.05). The means of serum FT3 and TT4 at presentation were correlated with the development of hypothyroidism after RAI therapy. Logistic regression analysis showed serum FT3 concentration at presentation to be significant contributing factor to failure to respond to a single dose of RAI. Patients who had higher FT3 concentrations at diagnosis were more likely to fail to respond to RAI therapy. CONCLUSION: The results of the present study of a cohort of patients with hyperthyroidism demonstrate that a single fixed dose of 10 mCi of RAI is highly effective in curing GD as well as toxic nodular hyperthyroidism. Therefore, treatment protocols for these groups should be identical. The most important factors that determine efficacy of RAI treatment are serum FT3 concentrations at diagnosis before the initiation of treatment and goiter size. Therefore, these factors should be taken into consideration when planning treatment. If such factors are present, the initial dose of RAI should be increased. PMID- 15545708 TI - Comparison of metabolic oscillations from mouse pancreatic beta cells and islets. AB - Rhythmic insulin secretion from pancreatic islets is the culmination of many processes both intrinsic and extrinsic to the beta cell. We wished to examine and compare endogenous metabolic oscillations in islets and isolated single beta cells that underlie secretion. Fluorescence patterns of rhodamine 123, an indicator of mitochondrial membrane potential (DeltaPsim), were analyzed for period and amplitude of oscillations using two methods: CLUSTER7 and fast Fourier transform (FFT). The period of DeltaPsim oscillations was greater in islets (271 +/- 21 s, n = 34) compared to dispersed beta cells (180 +/- 10 s, n = 54) by FFT analysis (p < 0.0005). CLUSTER7 confirmed differences in period and also detected oscillatory amplitude differences between beta cells (12.0 +/- 0.8) and islets (7.1 +/- 2.3% of baseline fluorescence, p < 0.0001). Depolarizing responses to the mitochondrial poison NaN3 were reduced in beta cells (35 +/- 4%) vs islets (58 +/- 9%), and hyperpolarizing responses to the calcium channel blocker nifedipine were enhanced in beta cells (15 +/- 4%) vs islets (8 +/- 1%), suggesting that mitochondria in dispersed beta cells were less energized than those in intact islets (p < 0.005), possibly due to elevated intracellular calcium. These findings suggest that individual beta cells possess the proper machinery to generate metabolic oscillations. Differences in oscillatory period, DeltaPsim, and nifedipine response suggest that incorporation into islets provides beta cells with additional modulatory influences. PMID- 15545710 TI - Conservation genomics: applying whole genome studies to species conservation efforts. AB - Studies of complete genomes are leading to a new understanding of the biology of mammals and providing ongoing insights into the fundamental aspects of the organization and evolution of biological systems. Comparison of primate genomes can identify aspects of their organization, regulation and function that appeared during the primate radiation, but without comparison to more evolutionarily distant mammals and other vertebrates, highly conserved aspects of genome architecture will not be accurately identified nor will the lineage-specific changes be identified as such. Many species of primates face risks of extinction; yet the knowledge of their genomes will provide a deeper understanding of primate adaptations, human origins, and provide the framework for discoveries anticipated to improve human medicine. The great apes, the closest relatives of the human species, are among the most vulnerable and most important for human medical studies. However, apes are not the only species whose genomic information will enrich humankind. Comparative genomic studies of endangered species can benefit conservation efforts on their behalf. Increased knowledge of genome makeup and variation in endangered species finds conservation application in population evaluation monitoring and management, understanding phylozoogeography, can enhance wildlife health management, identify risk factors for genetic disorders, and provide insights into demographic management of small populations in the wild and in captivity. PMID- 15545711 TI - Evolution of hominoids and the search for a genetic basis for creating humanness. AB - The phylogenetic relationship of human and apes are reviewed. The history of molecular phylogenetic studies in this field is then discussed, as is the role of natural selection at the molecular level. It is argued that approximately 10,000 genetic changes are responsible for creating human specific phenotypes. A genome wide comparison is necessary to decipher those changes. PMID- 15545712 TI - The dynamic nature and evolutionary history of subtelomeric and pericentromeric regions. AB - The organization and evolution of the subtelomeric and pericentromeric regions of human chromosomes exhibit unique characteristics compared to other regions of the genome. As shown in Fig. 1 the functional elements of the centromere and telomere are comprised of highly repetitive DNA sequences, which are responsible for carrying out the main mechanistic duties of these two regions: chromosome segregation and end replication, respectively. The nature of the repeats in these two regions and their function have been reviewed separately and, therefore, will not be discussed in more detail here (Sullivan et al., 1996, 2001; McEachern et al., 2000; Henikoff et al., 2001). Adjacent to these functional element regions, the centromere and telomere regions share an interesting architecture as depicted in Fig. 1. For both pericentromeric and subtelomeric regions, blocks of recent genomic duplications form a zone of shared sequence homologies between certain subsets of human chromosomes. The dynamic nature and evolutionary history of these regions and the unique DNA sequence adjacent to them will be the focus of this review. PMID- 15545713 TI - Primate phylogeny: molecular evidence from retroposons. AB - In these postgenomic times where aspects of functional genetics and character evolution form a focal point of human-mouse comparative research, primate phylogenetic research gained a widespread interest in evolutionary biology. Nevertheless, it also remains a controversial subject. Despite the surge in available primate sequences and corresponding phylogenetic interpretations, primate origins as well as several branching events in primate divergence are far from settled. The analysis of SINEs - short interspersed elements - as molecular cladistic markers represents a particularly interesting complement to sequence data. The following summarizes and discusses potential applications of this new approach in molecular phylogeny and outlines main results obtained with SINEs in the context of primate evolutionary research. Another molecular cladistic marker linking the tarsier with the anthropoid primates is also presented. This eliminates any possibility of confounding phylogenetic interpretations through lineage sorting phenomena and makes use of a new point of view in settling the phylogenetic relationships of the primate infraorders. PMID- 15545714 TI - The primates of the Neotropics: genomes and chromosomes. AB - The classification of neotropical primates has been controversial. Different arrangements have been proposed, depending on taxonomic criteria and on the traits selected for phylogenetic reconstructions. These include gross morphologic characters, karyotypic attributes and DNA sequence data of nuclear and mitochondrial genes and of repetitive genomic components. These approaches have substantially clarified the main intergeneric relationships although several intrageneric arrangements still remain to be elucidated. In this review, we compare karyologic and molecular data of this speciose group. PMID- 15545715 TI - Evolutionary implications of pericentromeric gene expression in humans. AB - Human pericentromeric sequences are enriched for recent sequence duplications. The continual creation and shuffling of these duplications can create novel intron-exon structures and it has been suggested that these regions have a function as gene nurseries. However, these sequences are also rich in satellite repeats which can repress transcription, and analyses of chromosomes 10 and 21 have suggested that they are transcript poor. Here, we investigate the relationship between pericentromeric duplication and transcription by analyzing the in silico transcriptional profiles within the proximal 1.5 Mb of genomic sequence on all human chromosome arms in relation to duplication status. We identify an approximately 5x excess of transcripts specific to cancer and/or testis in pericentromeric duplications compared to surrounding single copy sequence, with the expression of >50% of all transcripts in duplications being restricted to these tissues. We also identify an approximately 5x excess of transcripts in duplications which contain large quantities of interspersed repeats. These results indicate that the transcriptional profiles of duplicated and single copy sequences within pericentromeric DNA are distinct, suggesting that pericentromeric instability is unlikely to represent a common route for gene creation but may have a disproportionate effect upon genes whose function is restricted to the germ line. PMID- 15545716 TI - Tandem insertions of Alu elements. AB - Alu elements are non-autonomous, non-LTR retroposons that represent the most abundant mobile elements in the human genome (1.1 x 10(6) copies/genome). They preferentially insert adjacent to existing Alu elements. It has been proposed that Alu elements utilize LINE-1 machinery for their retroposition. The LINE-1 endonuclease cleaves at a loose consensus sequence. We have utilized a bioinformatics approach to show the order of insertion of pairs of young (Y) and old (S or J) Alu subfamily members. Our data suggest that the consensus LINE-1 endonuclease cleavage site used for insertion of the old Alu elements can be reused for integration of the younger ones inserting adjacent to them. However, there is also a preference at the 3' end of Alu into a non-ideal cleavage site that may represent unique properties of the A-tail for integration. Alu elements inserting adjacent to one another may suggest the saturation of the optimal integration sites with existing Alu elements, rather than any innate preference for Alu elements to integrate adjacent to other Alus. PMID- 15545717 TI - Identity by descent and DNA sequence variation of human SINE and LINE elements. AB - To test the hypothesis that Alu and L1 elements are genetic characters that are essentially homoplasy-free, we sequenced a total of five human L1 elements and eleven recently integrated Alu elements from 160 chromosomes (80 individuals representing four diverse human populations). Analysis of worldwide samples at L1 loci revealed 292 segregating sites and a nucleotide diversity of 0.0050. For Ya5 Alu loci, there were 129 segregating sites and nucleotide diversity was estimated at 0.0045. The Alu and L1 sequence diversity varied element to element. No completely or partially deleted Alu or L1 alleles were identified during the analysis. These data suggest that mobile element insertions are identical by descent characters for the study of human population genetics. PMID- 15545718 TI - Molecular evolution of the human chromosome 15 pericentromeric region. AB - We present a detailed molecular evolutionary analysis of 1.2 Mb from the pericentromeric region of human 15q11. Sequence analysis indicates the region has been subject to extensive interchromosomal and intrachromosomal duplications during primate evolution. Comparative FISH analyses among non-human primates show remarkable quantitative and qualitative differences in the organization and duplication history of this region - including lineage-specific deletions and duplication expansions. Phylogenetic and comparative analyses reveal that the region is composed of at least 24 distinct segmental duplications or duplicons that have populated the pericentromeric regions of the human genome over the last 40 million years of human evolution. The value of combining both cytogenetic and experimental data in understanding the complex forces which have shaped these regions is discussed. PMID- 15545719 TI - Nucleotide sequence comparison of a chromosome rearrangement on human chromosome 12 and the corresponding ape chromosomes. AB - Chromosome rearrangement has been considered to be important in the evolutionary process. Here, we demonstrate the evolutionary relationship of the rearranged human chromosome 12 and the corresponding chromosome XII in apes (chimpanzee, bonobo, gorilla, orangutan, and gibbon) by examining PCR products derived from the breakpoints of inversions and by conducting shotgun sequencing of a gorilla fosmid clone containing the breakpoint and a "duplicated segment" (duplicon). We confirmed that a pair of 23-kb duplicons flank the breakpoints of inversions on the long and short arms of chimpanzee chromosome XII. Although only the 23-kb duplicon on the long arm of chimpanzee chromosome XII and its telomeric flanking sequence are found to be conserved among the hominoids (human, great apes, and gibbons), the duplicon on the short arm of chimpanzee chromosome XII is suggested to be the result of a duplication from that on the long arm. Furthermore, the shotgun sequencing of a gorilla fosmid indicated that the breakpoint on the long arm of the gorilla is located at a different position 1.9 kb from that of chimpanzee. The region is flanked by a sequence homologous to that of human chromosome 6q22. Our findings and sequence analysis suggest a close relationship between segmental duplication and chromosome rearrangement (or breakpoint of inversion) in Hominoidea. The role of the chromosome rearrangement in speciation is also discussed based on our new results. PMID- 15545720 TI - Breakpoint analysis of the pericentric inversion between chimpanzee chromosome 10 and the homologous chromosome 12 in humans. AB - During this study, we analysed the pericentric inversion that distinguishes human chromosome 12 (HSA12) from the homologous chimpanzee chromosome (PTR10). Two large chimpanzee-specific duplications of 86 and 23 kb were observed in the breakpoint regions, which most probably occurred associated with the inversion. The inversion break in PTR10p caused the disruption of the SLCO1B3 gene in exon 11. However, the 86-kb duplication includes the functional SLCO1B3 locus, which is thus retained in the chimpanzee, although inverted to PTR10q. The second duplication spans 23 kb and does not contain expressed sequences. Eleven genes map to a region of about 1 Mb around the breakpoints. Six of these eleven genes are not among the differentially expressed genes as determined previously by comparing the human and chimpanzee transcriptome of fibroblast cell lines, blood leukocytes, liver and brain samples. These findings imply that the inversion did not cause major expression differences of these genes. Comparative FISH analysis with BACs spanning the inversion breakpoints in PTR on metaphase chromosomes of gorilla (GGO) confirmed that the pericentric inversion of the chromosome 12 homologs in GGO and PTR have distinct breakpoints and that humans retain the ancestral arrangement. These findings coincide with the trend observed in hominoid karyotype evolution that humans have a karyotype close to an ancestral one, while African great apes present with more derived chromosome arrangements. PMID- 15545721 TI - Genomic structure and paralogous regions of the inversion breakpoint occurring between human chromosome 3p12.3 and orangutan chromosome 2. AB - Intrachromosomal duplications play a significant role in human genome pathology and evolution. To better understand the molecular basis of evolutionary chromosome rearrangements, we performed molecular cytogenetic and sequence analyses of the breakpoint region that distinguishes human chromosome 3p12.3 and orangutan chromosome 2. FISH with region-specific BAC clones demonstrated that the breakpoint-flanking sequences are duplicated intrachromosomally on orangutan 2 and human 3q21 as well as at many pericentromeric and subtelomeric sites throughout the genomes. Breakage and rearrangement of the human 3p12.3-homologous region in the orangutan lineage were associated with a partial loss of duplicated sequences in the breakpoint region. Consistent with our FISH mapping results, computational analysis of the human chromosome 3 genomic sequence revealed three 3p12.3-paralogous sequence blocks on human chromosome 3q21 and smaller blocks on the short arm end 3p26-->p25. This is consistent with the view that sequences from an ancestral site at 3q21 were duplicated at 3p12.3 in a common ancestor of orangutan and humans. Our results show that evolutionary chromosome rearrangements are associated with microduplications and microdeletions, contributing to the DNA differences between closely related species. PMID- 15545722 TI - Cytochrome b polymorphisms and population structure of two species of Alouatta (Primates). AB - We carried out a phylogenetic and population study in Alouatta caraya and Alouatta belzebul based on cytochrome b DNA sequence data. Maximum Parsimony and Median-Joining analyses grouped A. caraya from different localities showing a population structure in accordance with geographic distribution. The relation between A. caraya haplotypes could be explained with respect to the species range in the Cerrado, one of the most ancient morphoclimatic domains of South America, and the Chaco. Conversely, A. belzebul from the Amazonas and Atlantic forests grouped in a paraphyletic arrangement without an evident geographic pattern. Recent geologic events resulting in the separation of A. belzebul might explain why these geographically distant groups shared similar haplotypes and why ancestral polymorphisms might have been maintained in this species. Time of divergence estimates indicated that the splitting of the Alouatta lineage leading to A. caraya occurred some 4.58 MYA while the lineage leading to A. belzebul emerged 4.14 MYA. PMID- 15545723 TI - The impact of chromosome sorting and painting on the comparative analysis of primate genomes. AB - Chromosome sorting by flow cytometry is the main source of chromosome-specific DNA for the production of painting probes. These probes have been used for cross species in situ hybridization in the construction of comparative maps, in the study of karyotype evolution and phylogenetics, in delineating territories in interphase nuclei, and in the analysis of chromosome breakpoints. We review here the contributions that this technology has made to the analysis of primate genomes. PMID- 15545724 TI - Origins of primate chromosomes - as delineated by Zoo-FISH and alignments of human and mouse draft genome sequences. AB - This review examines recent advances in comparative eutherian cytogenetics, including Zoo-FISH data from 30 non-primate species. These data provide insights into the nature of karyotype evolution and enable the confident reconstruction of ancestral primate and boreo-eutherian karyotypes with diploid chromosome numbers of 48 and 46 chromosomes, respectively. Nine human autosomes (1, 5, 6, 9, 11, 13, 17, 18, and 20) represent the syntenies of ancestral boreo-eutherian chromosomes and have been conserved for about 95 million years. The average rate of chromosomal exchanges in eutherian evolution is estimated to about 1.9 rearrangements per 10 million years (involving 3.4 chromosome breaks). The integrated analysis of Zoo-FISH data and alignments of human and mouse draft genome sequences allow the identification of breakpoints involved in primate evolution. Thus, the boundaries of ancestral eutherian conserved segments can be delineated precisely. The mapping of rearrangements onto the phylogenetic tree visualizes landmark chromosome rearrangements, which might have been involved in cladogenesis in eutherian evolution. PMID- 15545725 TI - Fluorescence in situ hybridization to chromosomes as a tool to understand human and primate genome evolution. AB - For the last 15 years molecular cytogenetic techniques have been extensively used to study primate evolution. Molecular probes were helpful to distinguish mammalian chromosomes and chromosome segments on the basis of their DNA content rather than solely on morphological features such as banding patterns. Various landmark rearrangements have been identified for most of the nodes in primate phylogeny while chromosome banding still provides helpful reference maps. Fluorescence in situ hybridization (FISH) techniques were used with probes of different complexity including chromosome painting probes, probes derived from chromosome sub-regions and in the size of a single gene. Since more recently, in silico techniques have been applied to trace down evolutionarily derived chromosome rearrangements by searching the human and mouse genome sequence databases. More detailed breakpoint analyses of chromosome rearrangements that occurred during higher primate evolution also gave some insights into the molecular changes in chromosome rearrangements that occurred in evolution. Hardly any "fusion genes" as known from chromosome rearrangements in cancer cells or dramatic "position effects" of genes transferred to new sites in primate genomes have been reported yet. Most breakpoint regions have been identified within gene poor areas rich in repetitive elements and/or low copy repeats (segmental duplications). The progress in various molecular and molecular-cytogenetic approaches including the recently launched chimpanzee genome project suggests that these new tools will have a significant impact on the further understanding of human genome evolution. PMID- 15545726 TI - Evolutionary conserved chromosomal segments in the human karyotype are bounded by unstable chromosome bands. AB - In this paper an ancestral karyotype for primates, defining for the first time the ancestral chromosome morphology and the banding patterns, is proposed, and the ancestral syntenic chromosomal segments are identified in the human karyotype. The chromosomal bands that are boundaries of ancestral segments are identified. We have analyzed from data published in the literature 35 different primate species from 19 genera, using the order Scandentia, as well as other published mammalian species as out-groups, and propose an ancestral chromosome number of 2n = 54 for primates, which includes the following chromosomal forms: 1(a+c(1)), 1(b+c(2)), 2a, 2b, 3/21, 4, 5, 6, 7a, 7b, 8, 9, 10a, 10b, 11, 12a/22a, 12b/22b, 13, 14/15, 16a, 16b, 17, 18, 19a, 19b, 20 and X and Y. From this analysis, we have been able to point out the human chromosome bands more "prone" to breakage during the evolutionary pathways and/or pathology processes. We have observed that 89.09% of the human chromosome bands, which are boundaries for ancestral chromosome segments, contain common fragile sites and/or intrachromosomal telomeric-like sequences. A more in depth analysis of twelve different human chromosomes has allowed us to determine that 62.16% of the chromosomal bands implicated in inversions and 100% involved in fusions/fissions correspond to fragile sites, intrachromosomal telomeric-like sequences and/or bands significantly affected by X irradiation. In addition, 73% of the bands affected in pathological processes are co-localized in bands where fragile sites, intrachromosomal telomeric-like sequences, bands significantly affected by X irradiation and/or evolutionary chromosomal bands have been described. Our data also support the hypothesis that chromosomal breakages detected in pathological processes are not randomly distributed along the chromosomes, but rather concentrate in those important evolutionary chromosome bands which correspond to fragile sites and/or intrachromosomal telomeric-like sequences. PMID- 15545727 TI - Reciprocal painting between humans, De Brazza's and patas monkeys reveals a major bifurcation in the Cercopithecini phylogenetic tree. AB - We report on reciprocal painting between humans and two Cercopithecini species, Erythrocebus patas (patas monkey) and Cercopithecus neglectus (De Brazza's monkey). Both human and monkeys chromosome-specific probes were made by degenerate oligonucleotide primed PCR (DOP-PCR) from flow sorted chromosomes. Metaphases of both monkey species were first hybridized with human chromosome specific probes and then human metaphases were hybridized with chromosome paints from each monkey species. The human paint probes detected 34 homologous segments on the C. neglectus karyotype, while the C. neglectus probes, including the Y, revealed 41 homologous segments on the human karyotype. The probes specific for human chromosomes detected 29 homologous segments in the E. patas karyotype, while the patas monkey probes painted 34 segments on the human karyotype. We tested various hypotheses of Cercopithecini phylogeny and taxonomy developed by morphologists, molecular biologists and cytogeneticists. Our hybridization data confirm that fissions (both Robertsonian and non-Robertsonian) are the main mechanism driving the evolutionary trend in Cercopithecini toward higher diploid numbers and strongly suggest an early phylogenetic bifurcation in Cercopithecini. One branch leads to Cercopithecus neglectus/Cercopithecus wolfi while the other line leads to Erythrocebus patas/Chlorocebus aethiops. Allenopithecus nigroviridis may have diverged prior to this major phylogenetic node. PMID- 15545728 TI - Phylogenetic inferences of Atelinae (Platyrrhini) based on multi-directional chromosome painting in Brachyteles arachnoides, Ateles paniscus paniscus and Ateles b. marginatus. AB - We performed multi-directional chromosome painting in a comparative cytogenetic study of the three Atelinae species Brachyteles arachnoides, Ateles paniscus paniscus and Ateles belzebuth marginatus, in order to reconstruct phylogenetic relationships within this Platyrrhini subfamily. Comparative chromosome maps between these species were established by multi-color fluorescence in situ hybridization (FISH) employing human, Saguinus oedipus and Lagothrix lagothricha chromosome-specific probes. The three species included in this study and four previously analyzed species from all four Atelinae genera were subjected to a phylogenetic analysis on the basis of a data matrix comprised of 82 discrete chromosome characters. The results confirmed that Atelinae represent a monophyletic clade with a putative ancestral karyotype of 2n = 62 chromosomes. Phylogenetic analysis revealed an evolutionary branching sequence [Alouatta [Brachyteles [Lagothrix and Ateles]]] in Atelinae and [Ateles belzebuth marginatus [Ateles paniscus paniscus [Ateles belzebuth hybridus and Ateles geoffroyi]]] in genus Ateles. The chromosomal data support a re-evaluation of the taxonomic status of Ateles b. hybridus. PMID- 15545729 TI - Investigation of marmoset hybrids (Cebuella pygmaea x Callithrix jacchus) and related Callitrichinae (Platyrrhini) by cross-species chromosome painting and comparative genomic hybridization. AB - We report on the cytogenetics of twin offspring from an interspecies cross in marmosets (Callitrichinae, Platyrrhini), resulting from a pairing between a female Common marmoset (Callithrix jacchus, 2n = 46) and a male Pygmy marmoset (Cebuella pygmaea, 2n = 44). We analyzed their karyotypes by multi-directional chromosome painting employing human, Saguinus oedipus and Lagothrix lagothricha chromosome-specific probes. Both hybrid individuals had a karyotype with a diploid chromosome number of 2n = 45. As a complementary tool, interspecies comparative genomic hybridization (iCGH) was performed in order to screen for genomic imbalances between the hybrids and their parental species, and between Callithrix argentata and S. oedipus, respectively. These genomic imbalances were confined to centromeric and telomeric heterochromatin, while euchromatic chromosome regions appeared balanced in all species investigated. When comparing marmosets and tamarins, sequence divergence of centromeric heterochromatin was already clearly noticeable. In the C. argentata and C. pygmaea genomes numerous subtelomeric regions were affected by amplification of different repetitive sequences. Cross-species FISH with a microdissection-derived C. pygmaea repetitive probe revealed species specificity of this repetitive sequence at the molecular cytogenetic level of resolution. PMID- 15545730 TI - Application of molecular cytogenetics for chromosomal evolution of the Lemuriformes (Prosimians). AB - R-banding chromosomal studies of 21 species of Lemuriformes allowed us to reconstruct the presumed ancestral karyotype of all the Lemuriformes except for Daubentoniidae and permitted the construction of their phylogenetic tree. Chromosome painting with fluorescently labeled heterologous DNA probes permitted comparative chromosome maps to be established. The Zoo-FISH method was used to reassess the karyotypes of 22 species or subspecies. While our results largely confirm the previous reconstruction of the ancestral karyotype, they resulted in a modification of the previously established phylogenetic tree. The Daubentoniidae emerged first followed by the divergence of the families Cheirogaleidae, Indriidae, Lepilemuridae and Lemuridae. Eight chromosome rearrangements occurred in all Lemuriformes except for Daubentoniidae in the common trunk. The present findings do not allow us to propose the occurrence of any rearrangement common to Daubentoniidae and other Lemuriformes, and probably other Prosimii. Conserved syntenies previously described in various mammalian orders were also conserved, while others were specific to the Lemuriformes. PMID- 15545731 TI - Evolutionary breakpoint analysis on Y chromosomes of higher primates provides insight into human Y evolution. AB - Comparative FISH mapping of PAC clones covering almost 3 Mb of the human AZFa region in Yq11.21 to metaphases of human and great apes unravels breakpoints that were involved in species-specific Y chromosome evolution. An astonishing clustering of evolutionary breakpoints was detected in the very proximal region on the long arm of the human Y chromosome in Yq11.21. These breakpoints were involved in deletions, one specific for the human and another for the orang-utan Y chromosome, in a duplicative translocation/transposition specific for bonobo and chimpanzee Y chromosomes and in a pericentric inversion specific for the gorilla Y chromosome. In addition, our comparative results allow the deduction of a model for the human Y chromosome evolution. PMID- 15545732 TI - The evolution of the azoospermia factor region AZFa in higher primates. AB - Clones of a PAC contig encompassing the human AZFa region in Yq11.21 were comparatively FISH mapped to great ape Y chromosomes. While the orthologous AZFa locus in the chimpanzee, the bonobo and the gorilla maps to the long arm of their Y chromosomes in Yq12.1-->q12.2, Yq13.1-->q13.2 and Yq11.2, respectively, it is found on the short arm of the orang-utan subspecies of Borneo and Sumatra, in Yp12.3 and Yp13.2, respectively. Regarding the order of PAC clones and genes within the AZFa region, no differences could be detected between apes and man, indicating a strong evolutionary stability of this non-recombining region. PMID- 15545733 TI - New insights into the evolution of chromosome 1. AB - A complex low-repetitive human DNA probe (BAC RP11-35B4) together with two microdissection-derived region-specific probes of the multicolor banding (MCB) probe-set for chromosome 1 were used to re-analyze the evolution of human chromosome 1 in comparison to four ape species. BAC RP11-35B4 derives from 1q21 and contains 143 kb of non-repetitive DNA; however, it produces three specific FISH signals in 1q21, 1p12 and 1p36.1 of Homo sapiens (HSA). Human chromosome 1 was studied in comparison to its homologues in Hylobates lar (HLA), Pongo pygmaeus (PPY), Gorilla gorilla (GGO) and Pan troglodytes (PTR). A duplication of sequences homologous to human 1p36.1 could be detected in PPY plus an additional signal on PPY 16q. The region homologous to HSA 1p36.1 is also duplicated in HLA, and split onto chromosomes 7q and 9p; the region homologous to HSA 1q21/1p12 is present as one region on 5q. Additionally, the breakpoint of a small pericentric inversion in the evolution of human chromosome 1 compared to other great ape species could be refined. In summary, the results obtained here are in concordance with previous reports; however, there is evidence for a deletion of regions homologous to human 1p34.2-->p34.1 during evolution in the Pongidae branch after separation of PPY. PMID- 15545734 TI - Panels of somatic cell hybrids specific for chimpanzee, gorilla, orangutan, and baboon. AB - The generation of panels of somatic cell hybrids specific for chimpanzee, gorilla, orangutan, and olive baboon is reported. The chromosome content of each hybrid clone was characterized using reverse painting on human normal metaphases and by the use of appropriate sequence tag sites (STSs), one for each chromosome arm. These resources can be advantageously exploited in the characterization of chromosome architecture of different primate species, with special reference to the discrimination of inter- and intra-chromosomal arrangement of segmental duplications. PMID- 15545735 TI - Comparative mapping of human claudin-1 (CLDN1) in great apes. AB - The gene encoding claudin-1 (CLDN1) has been mapped to human chromosome 3 (HSA3; 3q28-->q29) using a radiation hybrid panel. Employing fluorescence in situ hybridization (FISH) we here show that a human P1-derived artificial chromosome (PAC) containing CLDN1 detects the orthologous sites in chromosomes of the great apes, chimpanzee, gorilla, and orangutan. Furthermore, the chromosomal position of CLDN1 was determined in mouse chromosomes by FISH. The position of fluorescent signals is confined to a single chromosomal site in both great apes and mouse and in each case maps to the chromosomal region that has conserved synteny with HSA3 (PTR2q28, GGO2q28, PPY2q38 and MMU16B1). Using a gene-specific probe our results are consistent with reports of the striking similarity of great ape and human genomes as illustrated previously by chromosome painting. PMID- 15545736 TI - Evolutionary breakpoints are co-localized with fragile sites and intrachromosomal telomeric sequences in primates. AB - The concentration of evolutionary breakpoints in primate karyotypes in some particular regions or chromosome bands suggests that these chromosome regions are more prone to breakage. This is the first extensive comparative study which investigates a possible relationship of two genetic markers (intrachromosomal telomeric sequences [TTAGGG]n, [ITSs] and fragile sites [FSs]), which are implicated in the evolutionary process as well as in chromosome rearrangements. For this purpose, we have analyzed: (a) the cytogenetic expression of aphidicolin induced FSs in Cebus apella and Cebus nigrivittatus (F. Cebidae, Platyrrhini) and Mandrillus sphinx (F. Cercopithecidae, Catarrhini), and (b) the intrachromosomal position of telomeric-like sequences by FISH with a synthetic (TTAGGG)n probe in C. apella chromosomes. The multinomial FSM statistical model allowed us to determinate 53 FSs in C. apella, 16 FSs in C. nigrivittatus and 50 FSs in M. sphinx. As expected, all telomeres hybridized with the probe, and 55 intrachromosomal loci were also detected in the Cebus apella karyotype. The chi(2) test indicates that the coincidence of the location of Cebus and Mandrillus FSs with the location of human FSs is significant (P < 0.005). Based on a comparative cytogenetic study among different primate species we have identified (or described) the chromosome bands in the karyotypes of Papionini and Cebus species implicated in evolutionary reorganizations. More than 80% of these evolutionary breakpoints are located in chromosome bands that express FSs and/or contain ITSs. PMID- 15545737 TI - Chimpanzee chromosomes: retrotransposable compound repeat DNA organization (RCRO) and its influence on meiotic prophase and crossing-over. AB - The terminal C-bands that are a specific feature of chimpanzee chromosomes were dissected using a molecular cytogenetic technique, PRINS, with primers for telomeric sequences, subterminal satellite, and retrotransposable elements (HERV K and -W). These DNA elements jointly formed a large block of retrotransposable compound repeat DNA organization (RCRO) at the terminal C-band regions of 30 chromosomes, and are also located at the centromeric regions of some chromosomes. Additionally, a block consisting of all members of the RCRO has transposed to the middle (q31.1) of the long arm of chromosome 6, and three members, the subterminal satellite and the two HERVs, have integrated into the proximal region (q14.4) of the long arm of chromosome 14. Terminal RCROs seem to induce and prolong the bouquet stage in meiotic prophase, and to affect chiasma formation, together with interstitial RCROs. It is also postulated that RCROs may cause a position effect to gene expression, resulting in gene silencing and/or late replication. PMID- 15545738 TI - Inter- and intra-specific gene-density-correlated radial chromosome territory arrangements are conserved in Old World monkeys. AB - Recently it has been shown that the gene-density correlated radial distribution of human 18 and 19 homologous chromosome territories (CTs) is conserved in higher primates in spite of chromosomal rearrangements that occurred during evolution. However, these observations were limited to apes and New World monkey species. In order to provide further evidence for the evolutionary conservation of gene density-correlated CT arrangements, we extended our previous study to Old World monkeys. They comprise the remaining species group to be analyzed in order to obtain a comprehensive overview of the nuclear topology of human 18 and 19 homologous CTs in higher primates. In the present study we investigated four lymphoblastoid cell lines from three species of Old World monkeys by three dimensional fluorescence in situ hybridization (3D-FISH): two individuals of Japanese macaque (Macaca fuscata), crab-eating macaque (Macaca fascicularis), and an interspecies hybrid individual between African green monkey (Cercopithecus aethiops) and Patas monkey (Erythrocebus patas). Our data demonstrate that gene poor human 18 homologous CTs are located preferentially close to the nuclear periphery, whereas gene-dense human 19 homologous CTs are oriented towards the nuclear center in all cell lines analyzed. The gene-density-correlated positioning of human 18 and 19 homologous CTs is evolutionarily conserved throughout all major higher primate lineages, despite chromosomal inversions, fusions, fissions or reciprocal translocations that occurred in the course of evolution in these species. This remarkable preservation of a gene-density correlated chromatin arrangement gives further support for a functionally relevant higher-order chromatin architecture. PMID- 15545739 TI - Fine structure and meiotic behaviour of the male multiple sex chromosomes in the genus Alouatta. AB - The meiotic cytology and fine structure of the sex multiples in males from two species of the genus Alouatta are presented and compared with descriptions from other species of this genus. As shown in pachytene by synaptonemal complex analysis and in metaphase I by spreading, there is a quadrivalent in male meiosis in A. caraya, which is formed by an X(1)X(2)Y(1)Y(2) complex, while in A. palliata there is a trivalent formed by an X(1)X(2)Y(1) complex. Chromosome painting with human probes shows that A. caraya sex multiples share the same components as those of A. seniculus sara and A. seniculus arctoidea. However, as shown here for A. palliata and by others in A. fusca, there are differences among the multiples of some species. It is shown that in this genus there are several varieties of sex multiples that share some features, and that the origin of these multiples is most probably a primitive development in the genus Alouatta. PMID- 15545740 TI - Genetic testing for deafness is here, but how do we do it? PMID- 15545742 TI - Genetic tests and their evaluation: can we answer the key questions? AB - The rapid pace of research in the field of genetics has already yielded many benefits. The development of new genetic tests is one such example. Before there can be widespread uptake of these tests they need to be evaluated to confirm the benefits of their use. The authors review some of the key features of the evaluation of diagnostic tests focusing on analytical and clinical validity. Test properties such as sensitivity, specificity, likelihood ratios, positive and negative predictive values, and how they relate to molecular genetic testing are discussed. Associated issues such as the concepts of disease definition, imperfect reference standards, and false positives are also explored. The authors suggest possible approaches to addressing some of the problems identified. PMID- 15545741 TI - Genetic polymorphisms and heart failure. AB - Heart failure is a complex clinical syndrome. There is evidence for a genetic contribution to the pathophysiology of heart failure. Considering the fundamental role of neurohormonal factors in the pathophysiology and progression of cardiac dysfunction and hypertrophy, variants of genes involved in this system are logical candidate genes in heart failure. In this report, genetic polymorphisms of the major neurohormonal systems in heart failure will be discussed. Studies on polymorphisms of the renin-angiotensin-aldosterone system (RAAS), adrenergic receptor polymorphisms, endothelin (receptor) polymorphisms, and a group of miscellaneous polymorphisms that may be involved in the development or phenotypic expression of heart failure will be reviewed. Research on left ventricular hypertrophy is also included. The majority of genetic association studies focused on the ACE I/D polymorphism. Initial genetic associations have often been difficult to replicate, mainly due to problems in study design and lack of power. Promising results have been obtained with genetic polymorphisms of the RAAS and sympathetic system. Considering the evidence so far, a modifying role for these polymorphisms seems more likely than a role of these variants as susceptibility genes. Besides the need for larger studies to examine the effects of single nucleotide polymorphisms and haplotypes, future studies also need to focus on the complexity of these systems and study gene-gene interactions and gene-environment interactions. PMID- 15545743 TI - Lipoprotein lipase locus and progression of atherosclerosis in coronary-artery bypass grafts. AB - PURPOSE: Our aim was to test whether polymorphisms in the lipoprotein lipase (LPL) gene were associated with the progression of atherosclerosis in grafts examined in the Post-Coronary Artery Bypass Graft Trial (Post-CABG Trial). METHODS: 843 subjects in the post-CABG trial were genotyped for the LPL-D9N, N291S, PvuII, (TTTA)n, and HindIII polymorphisms. Associations between genotype and angiographically measured progression of atherosclerosis in grafts, medical history, and family history were examined. RESULTS: Greater progression of atherosclerosis was observed in subjects with LPL-HindIII 2/2 (56% versus 42% of those with other LPL HindIII genotypes, P = 0.025) and with LPL (TTTA)n 4/4 (63% versus 43% of those with other (TTTA)n genotypes, P = 0.020). Mantel-Haenszel analysis yielded an odds ratio of 1.84 for the effect of LPL HindIII 2/2 genotype on the progression of atherosclerosis in grafts (P = 0.015) and demonstrated that the effect of genotype on progression was of the same magnitude as, but independent of, the effect of drug treatment. CONCLUSION: The LPL-HindIII 2/2 genotype is a marker for genetic variation in the 3'-end of LPL that acts as an independent risk factor for the progression of atherosclerosis in grafts examined in the Post-CABG Trial. PMID- 15545744 TI - Maternal meiosis II nondisjunction in trisomy 21 is associated with maternal low socioeconomic status. AB - PURPOSE: We evaluated whether the association of socioeconomic risk factors for trisomy 21 differed by type of maternal meiotic error. METHODS: We determined meiotic errors by DNA analysis for 150 trisomy 21 cases, and maternal lifetime exposures to low socioeconomic factors by questionnaire. RESULTS: Mothers of meiosis II cases were significantly more likely to be exposed to four low socioeconomic factors than mothers of meiosis I cases (odds ratio = 9.50; 95% confidence interval = 1.8-49.8). CONCLUSION: Maternal lifetime exposure to poor socioeconomic environment is a risk factor for a trisomy 21, particularly if nondisjunction leads to a maternal meiosis II. PMID- 15545745 TI - Radiological features in 82 patients with nevoid basal cell carcinoma (NBCC or Gorlin) syndrome. AB - PURPOSE: Nevoid Basal Cell Carcinoma Syndrome (NBCCS) is an autosomal-dominant disorder characterized by multiple basal cell carcinomas, jaw cysts, palmar/plantar pits, calcification of the falx cerebri, and spine and rib anomalies. NBCCS is due to mutations in PTCH1, the human homologue of the Drosophila segment polarity gene patched. Mutations are detected in approximately 60% to 85% of individuals tested by sequencing of PTCH1; therefore, clinical examination and x-rays remain important in diagnosis of NBCCS. METHODS: We studied 82 NBCCS patients and 38 of their unaffected siblings at the NIH between 1985 and 1994. Chest, rib, spine, skull, hand and foot x-rays, brain MRI or CT, and pelvic ultrasound (in females) were obtained in the affected individuals and compared to their unaffected relatives. RESULTS: The following features were significantly more frequent in those with NBCCS: calcification of the falx cerebri, the most frequent radiological feature, was present in 79% of patients > 20 years and in 37% <20 years, calcification of the tentorium cerebellum was present in 20%, bridging of the sella in 68%, and abnormal frontal sinus aeration in 18% of affected individuals. Bifid ribs most often involving the third, fourth, and fifth ribs were seen in 26%; splayed, fused, and misshapen ribs in a further 16%, and widened ends of clavicles in 12%. Spine X-rays revealed calcification of the nuchal ligament in 18%, fusion of vertebrae in 10%, and hemivertebrae in 15%. Flame-shaped lucencies of the metacarpals and/or phalanges were present in 30%, modeling deformities of the phalanges in 14%, and polydactyly of the feet in 4%. The frequency of scoliosis, cervical ribs, absent or rudimentary ribs, spina bifida occulta, or short 4th metacarpal was not higher in the affected individuals as compared to their unaffected relatives. Except for falx calcification, the frequency of radiological manifestations was similar in different age groups. Cranial CT or MRI in 42 affected individuals revealed asymmetric or dilated ventricles in 24%, cerebral atrophy in 10%, cavum septum pellucidum in 19%, dysgenesis or agenesis of the corpus callosum in 10%, and meningioma in 5%. Ovarian fibromas were detected in 17% of females. CONCLUSIONS: This study reports the varied radiological manifestations of NBCCS. In the absence of major features such as basal cell carcinomas, jaw cysts, or falx calcification, which is often not evident until the teen years, other radiological manifestations of the disorder can permit early diagnosis of NBCCS in childhood. This will allow optimum surveillance for medulloblastoma and other neoplasms (cardiac fibromas and basal cell carcinomas) associated with NBCCS. PMID- 15545746 TI - Family communication about positive BRCA1 and BRCA2 genetic test results. AB - PURPOSE: The identification of a BRCA1 or BRCA2 genetic mutation can provide important health information to individuals who receive this result, but it can also provide crucial cancer risk information to family members. Most of the research on communication of genetic test results has focused on first degree relatives. The purpose of this retrospective study was to examine the process of communicating a positive BRCA1 or BRCA2 genetic test result to male and female first, second, and third degree relatives. METHODS: Participants were 38 female mutation carriers who responded to a written survey assessing the number and relationship of relatives informed, methods used to inform relatives, topics discussed, and motivations and barriers for communication. RESULTS: Overall, 59% (470/803) of first, second, and third degree relatives were informed. The proportion of informed parents, siblings, and offspring was nearly twice that of more distant relatives including nieces, nephews, aunts, uncles, grandchildren, and cousins (88% versus 45%; P = 0.02). The method of communication differed by the gender of the relative, as did some of the topics discussed. The most important reasons for discussing the genetic test results were (1) to inform the relatives of their risk, (2) to suggest that they be tested, and (3) to fulfill a perceived duty to inform. The major barrier to communication was little contact and/or emotionally distant relationships. CONCLUSION: Female mutation carriers act on a perceived duty to inform close relatives of their positive test result; however, there is a need for genetic counseling strategies that address communication with more distant relatives. PMID- 15545747 TI - Perceptions of genetic discrimination among at-risk relatives of colorectal cancer patients. AB - PURPOSE: To explore the concerns of at-risk relatives of colorectal cancer patients about genetic discrimination and their awareness of current legislative protections. METHODS: A questionnaire was sent to unaffected individuals with a family history of colorectal cancer who had enrolled in the Johns Hopkins Hereditary Colorectal Cancer Registry (N = 777). RESULTS: Of the 470 respondents, approximately half rated their level of concern about genetic discrimination as high. The majority of respondents, 79%, learned about genetic discrimination from at least one media source (television, newspapers, magazines, and radio). If they were to pursue genetic testing, respondents with a higher level of concern about genetic discrimination would be significantly more likely to pay out of pocket, use an alias, or ask for test results to be excluded from their medical record. Awareness and understanding of legislation regarding genetic discrimination was found to be minimal. CONCLUSION: Findings from this study demonstrate the negative effect of concerns about genetic discrimination on decisions about utilization of genetic services. Stronger legislative protections against genetic discrimination and increased public education through the scientific community and media sources are needed. PMID- 15545748 TI - Familial 22q11.2 deletions in DiGeorge/velocardiofacial syndrome are predominantly smaller than the commonly observed 3Mb. AB - PURPOSE: DiGeorge/velocardiofacial syndrome (DG/VCFS) is the most common cytogenetically characterized microdeletion of 22q11.2 region. In approximately 90% of patients, the deletion size is 3 Mb, whereas the remaining range from 1.5 to 2.5 Mb. The purpose of this study was to test the hypothesis that small deletions may be more easily tolerated in a familial fashion than larger deletions, especially for this syndrome. METHOD: Sixteen FISH probes designed from bacterial artificial chromosomes (BACs) and P1 artificial chromosomes (PACs) mapped to 22q11.2 were used to determine the deletion sizes in 22 individuals from ten families with familial 22q11.2 deletion detected by standard FISH tests. RESULT: Seven families had deletions of < 3 Mb ( approximately 1.5 Mb) in size and 3 families had the common 3-Mb deletion. The 70% frequency of smaller sized deletions among this group of patients with familial del(22)(q11.2) is significantly higher than that reported among unselected group of patients with del(22)(q11.2) (P < 0.0001, Fisher exact test). CONCLUSION: Familial del(22)(q11.2) are predominantly smaller than the common deletion size of 3 Mb, indicating that there may be some underlying mechanisms that favor parent-to child transmission of smaller deletions in individuals with del(22)(q11.2), therefore, underscoring the need to exclude a familial basis in cases of del(22)(q11.2) smaller than 3 Mb. PMID- 15545749 TI - Genetic testing as part of the early hearing detection and intervention (EHDI) process. PMID- 15545750 TI - Safeguards in the use of DNA databanks in genomic research. PMID- 15545751 TI - Nevoid basal cell carcinoma (Gorlin) syndrome. PMID- 15545752 TI - Second trimester maternal serum screening for fetal open neural tube defects and aneuploidy. PMID- 15545753 TI - Does providing hereditary breast cancer risk assessment support to practicing physicians decrease the likelihood of them discussing such risk with their patients? PMID- 15545756 TI - 2004 Founders Award. Presented to Kenneth L. Miller at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545754 TI - Outline of a medical genetics curriculum for internal medicine residency training programs. AB - To keep pace with the rapid advances in medical genetics, internal medicine residency training programs need to train internists to develop new attitudes, knowledge bases, and skill sets. Currently, such programs have no medical genetics curriculum. Thus, to set a minimum standard for genetics education in the context of training in internal medicine, the Internal Medicine Residency Training Program Genetics Curriculum Committee was formed, with members representing professional organizations of medical geneticists, internists, genetic counselors, internal medicine and genetics residency program directors, and internal medicine residents. The committee's task was to develop a concise outline of a medical genetics curriculum for residents in internal medicine in accordance with requirements of the Residency Review Committee for Internal Medicine of the Accreditation Council for Graduate Medical Education. The curriculum outline was drafted and circulated for comment. Before publication, the final document was approved by those member organizations that had a policy of approving curricula. Key learning objectives of the curriculum include appreciation of the rapid advances in genetics, the need for lifelong learning, the need for referral, and the role of genetic counselors and medical geneticists, as well as developing the ability to construct and analyze a three generation pedigree. A wide variety of teaching methods can be useful in these regards, including didactic lectures, multimedia CD- ROMs, and clinical experience. Teaching should be related to clinical experiences whenever possible. The curriculum developed by the committee and presented in this article will assist in teaching residents the attitudes, knowledge, and skills they will require. PMID- 15545757 TI - 2004 Elda E. Anderson Award. Presented to Timothy A. DeVol at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545758 TI - 2004 Distinguished Scientific Achievement Award. Presented to F. Ward Whicker at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545759 TI - 2004 Distinguished Scientific Achievement Award. Presented to Ralph H. Thomas at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545760 TI - 2004 Outstanding Science Teacher Award. Presented to John Shaefers at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545761 TI - 2004 Robley D. Evans Commemorative Medal. Presented to Geoffrey G. Eichholz at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545763 TI - 2004 William A. McAdams Outstanding Service Award. Presented to Edward F. Maher at the 49th annual meeting of the Health Physics Society, Washington, DC 11-15 July 2004. PMID- 15545765 TI - Test of CAP88-PC's predicted concentrations of tritium in air at Lawrence Livermore National Laboratory. AB - Based on annual tritium release rates from the five sources of tritium at Lawrence Livermore National Laboratory and the Tritium Research Laboratory at Sandia National Laboratory, the regulatory dispersion and dose model, CAP88-PC, was used to predict tritium concentrations in air at perimeter and offsite air surveillance monitoring locations for 1986 through 2001. These predictions were compared with mean annual measured concentrations, based on biweekly sampling. Deterministic predictions were compared with deterministic observations using predicted-to-observed ratios. In addition, the uncertainty on observations and predictions was assessed: when the uncertainty bounds of the observations overlapped with the uncertainty bounds of the predictions, the predictions were assumed to agree with the observations with high probability. Deterministically, 54% of all predictions were higher than the observations, and 96% fell within a factor of three. Accounting for uncertainty, 75% of all predictions agreed with the observations; 87% of the predictions either matched or exceeded the observations. Predictions equaled or exceeded observations at those sampling locations towards which the wind blows most frequently, except those in the hills. Under-predictions were seen at locations towards which the wind blows infrequently when released tritium was from elevated sources. When a high fraction of tritium was from area (diffuse) sources, predictions matched observations. PMID- 15545766 TI - Characterization of plutonium aerosol collected during an accident. AB - This study determined the plutonium particle size distribution and dissolution rate of PuO2 aerosol collected during the 16 March 2000 release of an undetermined amount of PuO2 in a room within a plutonium facility at Los Alamos National Laboratory. The facility has been in operation since 1978 to support the development, fabrication, and testing of Pu heat sources for the U.S. Department of Energy. Several workers were in the room at the time of the release and in vivo study of five of the workers began the day after the exposure event. Four of the subjects subsequently received chelation therapy. Over 30 fixed air filter samplers (FASs) and four continuous air monitors (CAMs) were operating in the room during the radiological release. One 47-mm-diameter glass fiber FAS filter and one 25-cm-diameter mixed cellulose ester CAM filter containing Pu aerosol from the incident were examined in the study described here. Total alpha radioactivity on the filters was determined by gross alpha counting. Isotopic identification of the Pu was made by alpha spectrometry. Film autoradiography was used to characterize the spatial distribution of alpha-emitting particles on the filters. Track-etch autoradiography was used to estimate the distribution of alpha radioactivity in individual plutonium particles on the filters for particle size measurement. The glass fiber filter was then cut into six sections. Particles from two sections were resuspended in alcohol, dispersed as an aerosol using a Lovelace nebulizer, and characterized by aerodynamic diameter using a Lovelace Multi-jet cascade impactor. The measured activity median aerodynamic diameter from the cascade impactor was 4.8 mum with a geometric standard deviation of 1.5. That agreed with the size distribution obtained from the alpha track detection technique. The remaining four filter sections were used in an in vitro dissolution study with synthetic serum ultrafiltrate. The retention of undissolved Pu was consistent with a biphasic exponential function. The majority of the Pu dissolved with a half-time of 900 d. The information on particle size distribution and solubility from this study was useful in assigning a radiation dose to the exposed workers, supporting the decision to administer chelation therapy, and providing a model for characterizing accident-associated aerosols in the future. PMID- 15545767 TI - Projecting the time trend of thyroid cancers: its impact on assessment of radiation-induced cancer risks. AB - The incidence of thyroid cancer, which may be induced by ionizing radiation, has been rising in most Western countries for more than 20 years. In France, public worry about this increase and its possible connection with the fallout from Chernobyl led the government to ask for an evaluation of the health impact of this accident and an assessment of the feasibility of an epidemiological study. These requests raise two methodological questions: Which risk model should be used to relate exposure to risk? What is known about the spontaneous incidence rate of thyroid cancers? This article analyzes the impact of the time trend in the spontaneous incidence of thyroid cancers over the past 20 years in France when evaluating the risk of radiation-induced cancer. Age-period-cohort models were used to model the trend of spontaneous incidence from 1978 through 1997 and then to apply two scenarios for projections up to 2007: one with a constant incidence, the other using the trend observed over the past 20 years. Then the risk was assessed for a hypothetical population of 30,000 children aged 0 to 15 y, exposed to a hypothetical 0.1 Gy thyroid dose. The analysis shows that consideration of the trend instead of a constant spontaneous incidence can yield substantial differences in the risk estimates for thyroid cancer. PMID- 15545768 TI - corneal epithelial injury thresholds for exposures to 1.54 microm radiation dependence on beam diameter. AB - Corneal epithelial injury thresholds have been determined for exposures to 1.54 mum infrared radiation from an Erbium fiber laser. Thresholds were determined for beam diameters from 0.05 to 0.7 cm for exposures having durations from approximately 1 to 100 s and for a fixed beam diameter of 0.1 cm for exposures with durations between 0.036 and 0.26 s. Near-threshold damage appeared within 30 min post-exposure. There was no evidence of latent damage from lesser exposures appearing up to 24-48 h post-exposure. The dependence of the threshold radiant exposures on laser beam diameter for exposures >1 s provides strong evidence supporting a critical temperature damage model. However, the shorter exposures are not in accord with a critical temperature damage model. Thresholds for exposures longer than 1 s are greater than 10 times the maximum permissible exposure (MPE) in ANSI Z-136.5-2000; however, the safety factor decreases to less than 10 for exposures less than 0.1 s with a 0.1-cm-diameter beam. PMID- 15545769 TI - The application of retrospective luminescence dosimetry in areas affected by fallout from the semipalatinsk nuclear test site: an evaluation of potential. AB - Luminescence retrospective dosimetry techniques have been applied with ceramic bricks to determine the cumulative external gamma dose due to fallout, primarily from the 1949 test, in populated regions lying NE of the Semipalatinsk Nuclear Test Site in Altai, Russia, and the Semipalatinsk region, Kazakhstan. As part of a pilot study, nine settlements were examined, three within the regions of highest predicted dose (Dolon in Kazakshstan; Laptev Log and Leshoz Topolinskiy in Russia) and the remainder of lower predicted dose (Akkol, Bolshaya Vladimrovka, Kanonerka, and Izvestka in Kazakshstan; Rubtsovsk and Kuria in Russia) within the lateral regions of the fallout trace due to the 1949 test. The settlement of Kainar, mainly affected by the 24 September 1951 nuclear test, was also examined. The bricks from this region were found to be generally suitable for use with the luminescence method. Estimates of cumulative absorbed dose in air due to fallout for Dolon and Kanonerka in Kazakshstan and Leshoz Topolinskiy were 475 +/- 110 mGy, 240 +/- 60 mGy, and 230 +/- 70 mGy, respectively. The result obtained in Dolon village is in agreement with published calculated estimates of dose normalized to Cs concentration in soil. At all the other locations (except Kainar) the experimental values of cumulative absorbed dose obtained indicated no significant dose due to fallout that could be detected within a margin of about 25 mGy. The results demonstrate the potential suitability of the luminescence method to map variations in cumulative dose within the relatively narrow corridor of fallout distribution from the 1949 test. Such work is needed to provide the basis for accurate dose reconstruction in settlements since the predominance of short-lived radionuclides in the fallout and a high degree of heterogeneity in the distribution of fallout are problematic for the application of conventional dosimetry techniques. PMID- 15545770 TI - Sample bottle design improvements for radon emanation analysis of natural waters. AB - The "radon emanation" method of analysis for Rn and Ra in natural waters has been used by scientists for more than 30 years. We have examined the use of a new plastic bottle design as a viable option and improvement to traditional glass designs. The advantages of the new design over that used previously in our laboratory include a larger volume, lower blank, less fragile nature, and ease of handling in the field. Extensive calibration of the new design yielded an average efficiency of 70.9 +/- 1.7% and showed that an accurate blank value can be determined for each sample based on its specific holding time. Based on a 5-L sample and a 60-min count time, minimum detectable activities (MDA) range from 1.0 to 1.2 mBq L for Rn and Ra, respectively. Memory effect experiments have shown that activity artifacts are negligible when reusing the sampling containers. In addition, intra- and inter-laboratory comparisons show that the new design gives reliable results. PMID- 15545771 TI - Residential radon exposure and lung cancer risk: commentary on Cohen's county based study. AB - The large United States county-based study () in which an inverse relationship has been suggested between residential low-dose radon levels and lung cancer mortality has been reviewed. While this study has been used to evaluate the validity of the linear nonthreshold theory, the grouped nature of its data limits the usefulness of this application. Our assessment of the study's approach, including a reanalysis of its data, also indicates that the likelihood of strong, undetected confounding effects by cigarette smoking, coupled with approximations of data values and uncertainties in accuracy of data sources regarding levels of radon exposure and intensity of smoking, compromises the study's analytic power. The most clear data for estimating lung cancer risk from low levels of radon exposure continue to rest with higher-dose studies of miner populations in which projections to zero dose are consistent with estimates arising from most case control studies regarding residential exposure. PMID- 15545773 TI - Uranium in hot water tanks: a source of TENORM. AB - Uranium deposits were detected inside hot water tanks using gamma-ray spectroscopic techniques and corroborated by the difference in the uranium concentration of the groundwater entering and leaving the hot water tanks. In situ gamma-ray spectroscopy was performed using a transportable high-purity germanium (HPGe) gamma-ray spectrometer to estimate the mass of uranium in the hot water tanks. Gamma-ray spectroscopic analyses of hot water tanks in four residences with groundwater uranium concentration between 732 and 7,667 mug L revealed an estimated 3.5 to 69 g of uranium in each hot water tank. The uranium deposit within the tanks was indicated by the 143.8, 163.4, and 185.7 keV gamma rays of U and confirmed with the 63.3, 92.3, and 92.8 keV gamma rays of Th as well as the 1,001 keV peak of Pa. An average decrease in uranium concentration of 23% was observed in the groundwater that passed through the hot water tanks. Additionally, once "uranium free" water entered the hot water tanks, the uranium deposits within the tanks resulted in an increase in the uranium concentration in the effluent water. The groundwater had an alkalinity in the range of 46-96 mg L as CaCO3 and a pH range of 7.3-8.1. The accumulation of uranium in these hot water tanks results in them being classified as technologically enhanced naturally occurring radioactive materials (TENORM). PMID- 15545774 TI - In situ depth profiling of 137Cs contamination in soils at the Idaho National Engineering and Environmental Laboratory. AB - Preremediation characterization of Cs contamination in soils was conducted at the Auxiliary Reactor Area (ARA)-23 Comprehensive Environmental Response, Compensation, and Liability Act site, located at the Idaho National Engineering and Environmental Laboratory (INEEL). Characterization activities included verification of the lateral extent of the contaminated area using the INEEL vehicle-mounted Global Positioning Radiometric Scanner. The vertical extent of the contamination in select areas of the site was evaluated with an in situ gamma ray spectrometer, and depth discrete samples were collected at 5-cm depth intervals down to a depth of 20.3 cm. A comparison was made between the depth distribution data from the in situ spectrometric measurements and the physical, depth discrete samples. The results of the study and of the aforementioned comparison indicate that use of in situ high-purity germanium (HPGe) detectors during the remediation of the ARA-23 site will aid in directing the depth of excavation, thereby helping to (a) minimize the amount of soils excavated and removed for disposal, and (b) reduce overall project costs. PMID- 15545775 TI - Danger in extrapolating indoor radon risk from underground miner data. PMID- 15545776 TI - Comment on "The U.S. Environmental Protection Agency's assessment of risks from indoor radon". PMID- 15545782 TI - Bruce Kessel, MD: 2004-2005 NAMS President. PMID- 15545783 TI - Development and clinical application of guidelines, consensus opinions, and position statements--the need for clinical judgment beyond the evidence. PMID- 15545784 TI - Can a blood test predict the onset of menopause? PMID- 15545785 TI - Modeling of antral follicle counts and reproductive aging. PMID- 15545786 TI - Recommendations for estrogen and progestogen use in peri-and postmenopausal women: October 2004 position statement of The North American Menopause Society. PMID- 15545787 TI - Anti-mullerian hormone is a promising predictor for the occurrence of the menopausal transition. AB - OBJECTIVE: Age at menopause and age at the start of the preceding period of cycle irregularity (menopausal transition) show considerable individual variation. In this study we explored several markers for their ability to predict the occurrence of the transition to menopause. DESIGN: A group of 81 normal women between 25 and 46 years of age visited the clinic two times (at T1 and T2) with an average interval of 4 years. All had a regular menstrual cycle pattern at T1. At T1, anti-mullerian hormone (AMH), follicle-stimulating hormone (FSH), inhibin B and estradiol (E2) were measured, and an antral follicle count (AFC) was made during the early follicular phase. At T2, information regarding cycle length and variability was obtained. Menopause transition was defined as a mean cycle length of less than 21 days or more than 35 days or as a mean cycle length of 21 to 35 days, but with the next cycle not predictable within 7 days during the last half year. A logistic regression analysis was performed, with the outcome measure as menopause transition. The area under the receiver operating curve (ROCAUC) was calculated as a measure of predictive accuracy. RESULTS: In 14 volunteers, the cycle had become irregular at T2. Compared with women with a regular cycle at T2, these women were significantly older (median 44.7 vs 39.8 y, P < 0.001) and differed significantly in AFC, AMH, FSH, and inhibin B levels assessed at T1. All parameters with the exception of E2 were significantly associated with the occurrence of cycle irregularity; AMH, AFC, and age had the highest predictive accuracy (ROCAUC 0.87, 0.80, and 0.82, respectively). After adjusting for age, only AMH and inhibin B were significantly associated with cycle irregularity. Inclusion of inhibin B and age to AMH in a multivariable model improved the predictive accuracy (ROCAUC 0.92). CONCLUSIONS: The novel marker AMH is a promising predictor for the occurrence of menopausal transition within 4 years. Adding inhibin B improved the prediction. Therefore, AMH alone or in combination with inhibin B may well prove a useful indicator for the reproductive status of an individual woman. PMID- 15545788 TI - Antral follicle counts are related to age at natural fertility loss and age at menopause. AB - OBJECTIVE: The variability in ultrasound-based antral follicle counts sized 2-10 mm after allowing for age-related decline is considerable. This may represent differences in actual reproductive age among women. This hypothesis was tested by cohort comparison for distribution of age at occurrence of reproductive events. DESIGN: A model with a nonlinear mean decline with age was fitted to antral follicle counts (AFC) obtained in 163 regularly cycling fertile volunteers. Ages at last child birth and menopause were predicted from the individual AFC by using thresholds to represent these events and the model for decline with age. Distributions of the observed ages at last childbirth (proxy variable for loss of natural fertility) and ages at menopause were obtained from the BALSAC demographic database and the Prospect-EPIC study, respectively. The observed distributions were compared with the predicted distributions by using visual comparison and quantile-quantile plots. Predictions of age at last child and age at menopause were done using percentiles of the modeled AFC distribution for given age, and corresponding percentiles of the predicted distributions of age at these reproductive events, with predictions following from the position of a woman's AFC relative to these percentiles. RESULTS: The predicted distributions of age at last child and age at menopause showed good agreement with the observed distributions in the BALSAC and EPIC cohort. Compared with age alone, antral follicle counts gave some additional information for individual prediction of age at last child and menopause. CONCLUSIONS: The link between declining antral follicle counts and reproductively significant events like loss of natural fertility and menopause is strengthened by the high degree of similarity among the predicted and observed age distributions. Predictive usefulness of this relationship in a clinical setting may be more marginal, except in the case of women who have low AFCs for their age. PMID- 15545789 TI - Osteoarthritis and osteoporosis in postmenopausal women: clinical similarities and differences. AB - Osteoarthritis and osteoporosis are two major health problems affecting more than 60% of post-menopausal women in the United States. The promotion of healthy aging and the prevention and reduction of morbidity and mortality is a main concern for healthcare providers. The similarities and differences in pathophysiology, diagnosis, and treatment for osteoarthritis and osteoporosis are often not clear for clinicians. Some osteoporosis treatments, including bisphosphonates and vitamin D, seem to have a beneficial effect on osteoarthritis as well. A review of these two conditions in terms of bone mineral density, bone turnover, hormonal effects, and treatment options will be discussed. PMID- 15545790 TI - Changes in bone density and turnover after alendronate or estrogen withdrawal. AB - OBJECTIVE: To compare bone mineral density (BMD) and bone turnover changes after therapy withdrawal in postmenopausal women treated with alendronate or estrogen progestin. DESIGN: In this randomized, blinded, multinational, placebo-controlled trial, 1,609 healthy postmenopausal women ages 45 to 59 years were assigned to receive alendronate, placebo, or open-label estrogen-progestin (conjugated equine estrogens plus medroxyprogesterone acetate or a cyclic regimen of 17 beta estradiol, norethisterone acetate and estradiol). Of the original women, one third after year 2 and one third after year 4 were switched from alendronate to placebo, while remaining blinded to treatment assignment. The women taking estrogen-progestin in years 1 to 4 were followed off therapy in years 5 and 6. BMD at the lumbar spine and hip and biochemical markers of bone turnover were measured. RESULTS: The treatment groups described in the current report represent 860 women at baseline; 481 women entered year 5, and 430 completed 6 years. BMD steadily decreased in the placebo group during all 6 years. In contrast, spine and hip BMD increased during the first 4 years in the groups receiving daily continuous alendronate 5 mg and estrogen-progestin. During years 5 and 6, BMD decreased at the lumbar spine -2.42% (95% CI = -4.10, -0.74) and total hip -1.09% (-2.60, 0.41) in the group previously treated with alendronate 5 mg for 4 years. In comparison, large BMD decreases were observed at the spine [-7.69% (-8.96, 6.41)] and total hip [-5.16% (-6.30, -4.01)] among women who had received estrogen-progestin for 4 years. CONCLUSION: Alendronate produces greater residual skeletal effects than estrogen-progestin after therapy discontinuation. PMID- 15545791 TI - Relation between climacteric symptoms and ovarian hypofunction in middle-aged and older Japanese women. AB - OBJECTIVE: To gain insight into the characteristics and current status of climacteric symptoms reported by middle-aged and older women in Japan, we surveyed women presenting at our menopause clinic. DESIGN: The participants included 1,069 women, ranging in age from 40 to less than 60 years (mean age, 50.2 y). Climacteric (indefinite) symptoms were objectively assessed with the use of the Keio questionnaire, which grades the severity of 40 types of symptoms classified into 20 subgroups. The total scores obtained for the 40 symptoms were used to calculate symptom prevalence and severity. To evaluate ovarian function, concentrations of estradiol and follicle-stimulating hormone (FSH) in sera were measured. RESULTS: The most frequent symptom was general fatigue, reported by 88.2% of the women. Shoulder stiffness was the symptom rated to be severe by the highest percentage of women (38.1%). The prevalence and severity of hot flushes (and sweats) were slightly higher in perimenopausal and early postmenopausal women than in premenopausal and late postmenopausal women. The prevalence and severity of hot flushes and sweats were higher in women with estradiol < 25 pg/mL and FSH > 40 mIU/mL than in those with estradiol > or = 25 pg/mL and FSH < or = 40 mIU/mL. CONCLUSION: General fatigue and shoulder stiffness, symptoms with low hormone dependence, are the two most frequent climacteric symptoms in our clinic. Hot flushes and sweats, symptoms with high hormone dependence, are also common symptoms. PMID- 15545793 TI - Comment on "Bioidentical hormone therapy: a review". PMID- 15545794 TI - Comment on "Bioidentical hormone therapy: a review". PMID- 15545797 TI - Guarding against vascular site infection. Arm yourself with the latest knowledge on equipment and technique to protect patients from catheter-related bloodstream infections. PMID- 15545799 TI - Know your hand in hygiene. PMID- 15545800 TI - Watch that incision! PMID- 15545801 TI - Hypothermia's chilling effect on outcomes. PMID- 15545802 TI - Cut down sterilization turnaround time. PMID- 15545803 TI - Preview the OR of the future. PMID- 15545807 TI - Normal eye growth in emmetropic schoolchildren. AB - PURPOSE: The purpose of this report is to describe the normal growth pattern of the optical components of the eye in a cohort of emmetropic, school-aged children. METHODS: Emmetropia was defined as refractive error (measured by cycloplegic autorefraction) in the vertical and horizontal meridians of the right eye between +1.00 D and -0.25 D at all the visits. This definition resulted in a sample of 194 children enrolled in the Orinda Longitudinal Study of Myopia (OLSM) between ages 6 and 14 years with at least 2 years of follow-up evaluation (across three annual visits) between 1989 and 2000. The optical components measured included corneal power, anterior chamber depth, crystalline lens thickness, Gullstrand lens power, calculated lens power, crystalline lens index, vitreous chamber depth, and axial length. RESULTS: Corneal power and anterior chamber depth were best modeled as quadratic functions of ln (age). The model involving the square of the inverse of age best described calculated lens power and crystalline lens index. The relationship between age and crystalline lens thickness was best described using a linear function of age with a point of inflection. A linear function of ln (age) with a point of inflection best described the relationship between age and axial length, Gullstrand lens power, and vitreous chamber depth. For five of the eight components (crystalline lens thickness, Gullstrand lens power, calculated lens power, corneal power, and crystalline lens index), the line modeling the data was negative in overall direction, indicating that the component value decreased with age. The upward trend of the line modeling axial length, anterior chamber depth, and vitreous chamber depth reflected the continued growth of the eye from age 6 years to age 15 years. CONCLUSIONS: A picture of normal eye growth in emmetropes from ages 6 to 15 years is provided based on a combination of cross-sectional and longitudinal data. Axial elongation, crystalline lens flattening and thinning, and decrease in lens power are its hallmarks. PMID- 15545808 TI - Repeatability of IOLMaster biometry in children. AB - AIMS: This study compares the repeatability of IOLMaster (Zeiss, Oberkochen, Germany) axial dimension measurements and conventional ultrasonography in children. METHODS: A series of IOLMaster (partial coherence interferometry, optical pachometry) and Echoscan (US 800, Nidek, Tokyo, Japan) (ultrasound) measurements were taken on 179 Chinese children (mean age, 10.6 +/- 0.8 years) taking part in a longitudinal study of myopia development, and the measurements were repeated on 37 of these subjects. RESULTS: IOLMaster axial length measurements showed better repeatability (95% limits of agreement for repeatability, -0.047 to 0.038 mm) than Echoscan axial length measurements (95% limits of agreement for repeatability, -0.85 to 0.67 mm). IOLMaster anterior chamber depth measurements also showed better repeatability (95% limits of agreement for repeatability, -0.053 to 0.073 mm) than Echoscan anterior chamber depth measurements (95% limits of agreement for repeatability, -0.57 to 0.49 mm). IOLMaster measurements were, on average, slightly larger than Echoscan measurements for axial length (by 0.14 mm) and anterior chamber depth (0.09 mm). CONCLUSION: Partial coherence interferometry techniques, such as that used by IOLMaster, should be considered as the standard technique for axial length measurement in children because they are noninvasive, highly precise, and easy to use. PMID- 15545809 TI - Gain and movement time of convergence-accommodation in preschool children. AB - BACKGROUND: Convergence-accommodation is the synkinetic change in accommodation driven by vergence. A few studies have investigated the static and dynamic properties of this cross-link in adults but little is known about convergence accommodation in children. The purpose of this study was to develop a technique for measuring convergence-accommodation and to study its dynamics (gain and movement time) in a sample of pre-school children. METHOD: Convergence accommodation measures were examined on thiry-seven normal pre-school children (mean age = 4.0 +/- 1.31 yrs). Stimulus CA/C (sCA/C) ratios and movement time measures of convergence-accommodation were assessed using a photorefractor while subjects viewed a DOG target. Repeated measures were obtained on eight normal adults (mean age = 23 +/- 0.2 yrs). RESULTS: The mean sCA/C ratios and movement times were not significantly different between adults and children (0.10 D/Delta [0.61 D/M.A.], 743 +/- 70 ms and 0.11 D/Delta [0.50 D/M.A.], 787 +/- 216 ms). Repeated measures on adults showed a non-significant mean difference of 0.001 D/Delta. CONCLUSION: The results suggest that the possible differences in crystalline lens (plant) characteristics between children and adults do not appear to influence convergence-accommodation gain or duration. PMID- 15545810 TI - Simulated impairment of contrast sensitivity: performance and gaze behavior during locomotion through a built environment. AB - PURPOSE: An experimental study investigated the effects of simulated impairment of contrast sensitivity (CS) on performance and eye gaze patterns during locomotion through a library. METHOD: Normally sighted participants with simulated CS impairment (diffusive blur) walked two routes, one entailing limited change in direction (simple) and the other entailing several changes in direction (complex), while eye movements relative to the scene were recorded. Performance variables included walking speed in completing the route, pauses during travel, and collisions with objects on the route. For eye movements, dwell time and saccades were determined for each of three object classes: (1) objects on the route below eye level; (2) objects on the route extending above eye level; and (3) elsewhere-objects not on the route. RESULTS: Walking speed was significantly affected by CS level and by route; pauses and collisions were rare. Dwell times and saccades suggested limited attention directed to low-level objects, except for CS impairment on the complex route. In the complex route, saccades and dwell times in the object class "elsewhere" were also reduced. CONCLUSIONS: The results for the simple route suggest ballistic strategies: the participant appraises the scene and follows a more or less predetermined path. For the complex route, CS impairment appears to adversely affect information processing and locomotion. The results have implications for the design of built environments, especially with regard to the safety of visually impaired occupants during emergency scenarios. PMID- 15545811 TI - Measurements of ocular aberrations and light scatter in healthy subjects. AB - PURPOSE: To report and validate an optical imaging system that provides measurements of higher order ocular aberrations and light scatter in human eyes. METHODS: An optical imaging system has been established that provides for combined measurements of ocular aberrations and light scatter. A laser beam was expanded and focused to a point on the retina by the optics of the eye. Wavefront sensing was performed with a Shack-Hartmann aberrometer to determine the wavefront aberration function and calculate the point spread function, giving information on ocular aberrations. A cylindrical lens was placed in the path of the incident laser beam path, and the line spread function was derived from the laser slit, giving information on combined ocular aberrations and light scatter. A relative index for ocular light scatter was determined by subtracting the area under the two line spread functions. Measurements were performed in one eye of 20 normal healthy subjects. The subjects' ages ranged between 21 and 78 years, and the average for all the eyes was 43 +/- 17 years (mean +/- SD). RESULTS: Higher order ocular aberrations were correlated with subjects' ages (r = 0.6; p = 0.01; N = 20). Combined higher order ocular aberrations and light scatter were correlated with age (r = 0.7; p = 0.0002; N = 20). Light scatter was correlated with age (r = 0.6; p = 0.002; N = 20). CONCLUSIONS: A method was established to measure age-related changes in ocular higher order aberrations and light scatter. Differentiating the contribution of ocular aberrations and light scatter to the retinal image quality has potential value for anticipating the outcome of procedures that attempt to compensate for ocular aberrations and for providing information on factors that degrade the optical performance of the eye in health and disease. PMID- 15545812 TI - Corneal endothelial cell morphometry and corneal thickness in diabetic contact lens wearers. AB - PURPOSE: To assess corneal endothelial cell morphometry and corneal thickness in diabetic patients who wear contact lenses. METHODS: Images of the central corneal endothelium were analyzed quantitatively and qualitatively and corneal thickness was measured in a group of diabetic patients (type 1, N = 26; type 2, N = 4) who wear soft contact lenses and in a group of nondiabetic age-matched control subjects who were also contact lens wearers. RESULTS: Endothelial cell characteristics and corneal thickness values were similar for the two groups (p > 0.05). Four of the diabetic patients (and none of the nondiabetic patients) displayed folds in the endothelial mosaic. CONCLUSIONS: The morphometry of corneal endothelial cells and central corneal thickness values in diabetic patients who wear soft contact lenses were not appreciably different from those found in lens-wearing control subjects. PMID- 15545813 TI - A novel low-order method for recovery of the corneal shape. AB - PURPOSE: To describe a novel low-order method for the recovery of the corneal height from videokeratographically obtained images. The method uses an iterative cubic approach with implicit continuous curvature. Convergence is easily established for a particular videokeratograph. The effect of skew rays is treated in a postprocessing step. METHOD: Four simulated model corneas are tested: (1) an asphere; (2) an ellipsoid; (3) a radially keratotomized cornea; and (4) a simulation of photorefractive keratectomy (PRK). The corneal height, slope, and tilt are compared against theory and a 2nd order Taylor series method using root mean square error measures. The effect of lateral and axial shifts (up to 0.1 mm) is examined. The two methods are tested (experimentally) on an 8-mm spherical calibration ball, in which the image data are processed using a least-squares calibration procedure. RESULTS: The lowest height errors are found for the asphere and PRK models (4.5 x 10 microm and 3.6 x 10 microm). The maximum height error is 0.38 microm (ellipsoid), with 0.14 microm average overall error and 0.45 microm error for the 8-mm calibration ball. The comparison method has an average error of 0.92 microm, with maximum error of 1.5 microm (PRK) and 0.55 microm error for the 8-mm calibration ball. Shift induces larger (approximately 200 microm/mm) height errors than axial shift (approximately 2 microm/mm), but the errors are similar between methods. A demonstration on additional ellipsoids suggests the new algorithm (without skew ray compensation) is not as effective as the comparison method for increasingly nonspherical surfaces. The method has a short completion time of 2.3 s in seven iterations using MATLAB version 5.0 (The MathWorks, Inc., Natick, MA), running on a Pentium III 667-MHz processor with 128 MB RAM, and running the Windows 2000 operating system (Microsoft, Redmond, WA). CONCLUSION: An accurate, simple, robust, iteratively stable and fast method for estimating the corneal shape is described. A recovery with continuous curvature and skew ray compensation suggests a potential improvement within the context of current standard corneal shape recovery algorithms. PMID- 15545814 TI - Critique of claims of improved visual acuity after hypnotic suggestion. AB - Psychological approaches to improving vision present an enticing alternative to invasive procedures and corrective lenses; hypnotic suggestion is one such technique. During the past 60 years, multiple studies have documented improvements in the vision of myopic individuals after hypnotic interventions. Given the increasing interest in behavioral and alternative approaches, we have reviewed the pertinent studies to evaluate their validity. We delineate various shortcomings in these reports, including potential methodological caveats, problems with experimental controls, and controversial data interpretation. Overall, the data do not seem to support hypnosis as a viable option for significant long-term improvement of myopia. However, hypnosis can increase one's subjective feeling of enhanced visual acuity by affecting higher cognitive functions, such as attention, memorization, and perceptual learning, which could influence performance on visual tasks. PMID- 15545815 TI - Tolerating vertex distance changes for spherocylindrical corrections. AB - Prescriptions depend on the vertex distance. Although weak prescriptions are insensitive to a vertex distance change, stronger ones must be adjusted if the vertex distance is modified by an appreciable amount. The tolerable amount can be specified for pure spherical powers. For spherocylindrical corrections, the concept of dioptric distance is invoked in this article and applied to the propagation of an astigmatic wavefront. This leads to a simple rule that is well suited to decide on the necessity of modifying a prescription. PMID- 15545816 TI - The recovery of posterior cornea and anterior lens radii by a novel ray-tracing method. AB - BACKGROUND: Methods for estimating ocular surface radii are typically based on paraxial vergence calculations, need to account for finite source positions, and require refocusing of the camera. This article describes (1) a telecentric ray finding method; and (2) its application to the problem of determining posterior cornea (R2) and anterior lens (R3) radii by a regression procedure that addresses these issues. METHODS: The ray-finding algorithm simulates Purkinje image heights Pj(h) (for j = 2, 3; for the Le Grand eye) for the expected range of Rj. A two step cubic regression procedure fits this image data globally and then over a refined interval to estimate Rj locally. The goodness of fit is measured by the R statistic. A standard method is compared with the new method in simulation. Mean absolute errors and SD's are recorded for 10 randomly chosen R2 and R3. The effect of errors caused by (1) axial shift of the posterior cornea and anterior lens (axially up to +/-0.1 mm); and (2) camera digitization (pixel sizes of 20 microm) are simulated. The method can make use of general surface height information; therefore, it is tested on an eye with nonspherical cornea shape (a toroidal surface). RESULTS: The time to generate Rj is no more than 45 s (R3), with R > 0.9999. The errors for the unmodified eye are (1.3 +/- 2.1) x 10 mm (R2) and (2.4 +/- 1.6) x 10 mm (R3) (new) vs. (5.6 +/- 0.9) x 10 mm (R2) and (1.1 +/- 1.0) x 10 mm (R3) (standard). Digitization increases errors to 0.32 +/- 0.16 mm (R2) and 0.10 +/- 0.10 (R3) (new) vs. 0.45 +/- 0.33 mm (R2) and 0.48 +/- 0.13 mm (R3) (standard). Axial shift error for R2 is similar between methods, with a tendency toward lower error for the new method given digitization error. This trend is found for R3, although the new method now does consistently better with digitization error. Shift contributes the smaller proportion of total error (approximately 10 mm) compared with digitization (approximately 10 mm). The errors for the toroidal surface are (5.7 +/- 7.6) x 10 mm (R2) and (1.1 +/- 0.7) x 10 mm (R3) (new) compared with errors of 0.18 +/- 0.01 mm (R2) and 0.44 +/- 0.07 mm (R3) (standard). The new method produces better results in this case. CONCLUSION: A telecentric image-computing algorithm produces accurate image positions. A two-step cubic regression produces accurate estimates of R2 and R3 (errors approximately 10 mm). This error increases with axial shift (approximately 10 mm) and digitization (approximately 10 mm). The new method does better than a standard method ignoring all the errors and tends to handle digitization error better. The new method works well on a toroidal anterior cornea. Testing on model/real eyes is required. Efforts are continuing to refine methods for videophakometry. PMID- 15545817 TI - Blind obedience or plain stupidity? PMID- 15545819 TI - Left ventricular contractile dysfunction as a complication of thermal injury. PMID- 15545820 TI - What can we learn from the three megatrials using anticoagulants in severe sepsis? PMID- 15545821 TI - CXC-chemokine stimulation of neutrophils correlates with plasma levels of myeloperoxidase and lactoferrin and contributes to clinical outcome after pediatric cardiac surgery. AB - Several CXC-chemokines, of which interleukin (IL)-8 is the prototype, are potent neutrophil chemotactic and activating cytokines, inducing the secretion of granule proteins and the generation of reactive oxygen intermediates that may cause tissue damage and amplify inflammatory responses. Here, we investigated whether chemokines play a key role in the inflammatory process following cardiac surgery with cardiopulmonary bypass (CPB) in children. We performed an observational prospective clinical study of 40 pediatric patients before, during, and after open heart surgery with CPB. Plasma levels of chemokines, myeloperoxidase (MPO), and lactoferrin were measured by immunoassays. Cell surface receptors were detected by flow cytometry. Plasma levels of IL-8 were increased after CPB, correlating strongly with a reduction of expression of the CXC-chemokine receptors (CXCR) 1 and 2 on neutrophils indicating in vivo activation of neutrophils by IL-8. Other CXC-chemokines with Glu-Leu-Arg motif showed no correlation with CXCR1 or CXCR2 expression. Two components of neutrophilic granules, MPO and lactoferrin, were strongly elevated postoperatively, and the levels of both were correlated with IL-8. Levels of monocyte chemoattractant protein (MCP)-1 were increased postoperatively, correlating with a reduction of CCR2 expression and an increase of CD11b expression on monocytes, suggesting monocyte activation by MCP-1. The early postoperative course was complicated in patients with an increase of these inflammatory parameters. Impaired cardiovascular function correlated with increased levels of IL-8 and activation of neutrophils and was most prominent in patients with a long time on CPB and in those with cyanotic heart lesions. In conclusion, MCP-1 is involved in the regulation of chemotaxis and function of monocytes during and early after the end of CPB. Activation of neutrophils and down-regulation of CXCR1 and CXCR2 were predominantly caused by IL-8. This activation implies release of components of neutrophilic granules and correlates with the need for inotropic support. PMID- 15545822 TI - Histocompatibility leukocyte antigen-D related expression is specifically altered and predicts mortality in septic shock but not in other causes of shock. AB - Although the expression of monocyte histocompatibility leukocyte antigen (HLA)-DR has been shown to be decreased during human sepsis, its level of expression in other nonseptic critical conditions is unclear. The aim of this study was to compare the level of HLA-DR expression on circulating monocytes among patients with septic, hemorrhagic, and cardiogenic shocks and severe sepsis without shock. At admission, HLA-DR expression was exclusively decreased in patients with septic shock (n = 30; P < 0.001), whereas the expression was similar between the other studied groups: cardiogenic shock (n = 16), hemorrhagic shock (n = 11), severe sepsis without shock (n = 18), and healthy volunteers (n = 8). HLA-DR expression was not predictive for overall mortality, but at day 1, an HLA-DR expression of less than 14 of mean fluorescence intensity (that corresponds to 40% labeled monocytes) was predictive of mortality exclusively in patients with septic shock (odds ratio, 11.4 and 95% confidence interval, 1.7; 78.4; P < 0.008). Catecholamine infusion, mechanical ventilation, positive blood culture, and number of units of blood or plasma transfused did not correlate with decreased HLA-DR expression. Thus, the decrease in HLA-DR expression is specific to septic shock and is associated, in septic shock patients, with increased mortality risk. PMID- 15545823 TI - Initial posttraumatic translocation of NF-kappaB and TNF-alpha mRNA expression in peripheral blood monocytes of trauma patients with multiple injuries: a pilot study. AB - Post-traumatic inflammation is connected to monocyte dysfunction characterized by reduced NF-kappaB translocation during the first post-traumatic days. Because the exact dynamic of monocytic NF-kappaB translocation in patients directly after trauma remains unclear, the aim of this pilot study was to measure the intranuclear presence of NF-kappaB in monocytes from patients with multiple injuries initially after the trauma and during the early post-traumatic period and to compare these results with downstream-placed mRNA expression alteration of TNF-alpha, as well as with clinical data. Eleven patients were enrolled with an Injury Severity Score of 16 to 66 points, and blood samples were drawn on admission within 90 min and at 6, 12, 24, 48, and 72 h after trauma. NF-kappaB translocation of monocytic nuclear protein was analyzed by electrophoretic mobility shift assay and was quantified by densitometry as arbitrary units. In addition, monocytes of healthy volunteers were analyzed either native (-, control) or after LPS stimulation (+, control). For determination of downstream mRNA encoding for TNF-alpha, quantitative reverse transcriptase-PCR was performed. For both parameters, the negative control values were set as baseline (=1) and results from positive controls and patients were given as a relative alteration ratio without unit. Initial post-traumatic NF-kappaB translocation was significantly increased in trauma patients on admission (88 +/- 37) and 6 h after trauma (59 +/- 28) compared with the baseline level. In contrast, TNF-alpha mRNA was not increased on admission (1.7 +/- 0.9) and decreased even below baseline after 12 h. The substantial information of our study arises from the analysis of the dynamic of NF-kappaB translocation of monocytes. Enabled by closely matched sequential blood sampling strictly standardized to the traumatic event, an essential increase of monocytic signal transduction and transcription could be elucidated in the very early post-traumatic period, which precedes the down regulation of the innate immune system. PMID- 15545824 TI - Circulating levels of macrophage migration inhibitory factor are associated with mild pulmonary dysfunction after cardiopulmonary bypass. AB - Macrophage migration inhibitory factor (MIF) is a central mediator of inflammatory response and acute lung injury that is secreted in response to corticosteroids. A rise in systemic MIF levels was described after cardiac surgery in steroid-treated patients. This study aimed to investigate the circulating levels of MIF and the possible relationship of this cytokine to pulmonary dysfunction after cardiopulmonary bypass (CPB). We included 74 patients without previous organ dysfunction undergoing elective coronary artery bypass surgery (CABS). The same team performed all CABS via a standard technique adding methylprednisolone (15 mg/kg) to the CPB priming solution (Group MP, n = 37). In the remaining patients (Group NS, n = 37), methylprednisolone was withdrawn from the CPB priming. MIF, C-reactive protein (CRP), and total C3 were assayed in peripheral blood sampled immediately before anesthesia induction and 3, 6, and 24 h post-CPB. Preoperative risk scores and peri- and postoperative variables were documented. Postoperative kinetics of MIF and C3 were similar for both groups. Levels of CRP 24 h post-CPB were higher in Group MP (P = 0.003). Higher MIF levels were detected 6 h post-CPB, and returned to preoperative levels 24 h after CPB. MIF levels 6 h post-CPB were inversely related to the postoperative PaO2/FiO2 ratio (P = 0.0021) and were directly related to the duration of mechanical ventilation (P = 0.014). Perioperative use of methylprednisolone did not modify the MIF response to CPB, but it was related to an enhanced acute phase response. Higher circulating MIF levels 6 h post-CPB were associated with worse postoperative pulmonary short-course outcome. PMID- 15545825 TI - Cd40 but not CD154 knockout mice have reduced inflammatory response in polymicrobial sepsis: a potential role for Escherichia coli heat shock protein 70 in CD40-mediated inflammation in vivo. AB - The CD40-CD154 system controls various aspects of the host inflammatory response in models of cellular and humoral immunity. Recently, we described a role for CD40 in the innate immune response in polymicrobial sepsis. However, recent data suggests that CD40 maybe activated by CD154 or directly via bacterial heat shock protein (HSP) 70. Therefore, we decided to test the mechanism of CD40 activation in murine polymicrobial sepsis. Wild-type (WT), CD40, and CD154 underwent cecal ligation and puncture (CLP). Compared with WT mice, CD40 had improved survival in association with attenuated production of IL-12, TNF-alpha, and IL-6. In contrast, CD154 mice behaved similar to WT mice with regard to mortality and cytokine production. The differential response of CD40 and CD154 mice to CLP was not due to a general attenuated response to inflammatory stimuli, as all three strains had similar survival after LPS administration, and CD40 macrophages had normal production of IL-12 in response to lipopolysaccharide. In contrast, CD40 macrophages had attenuated IL-12 production in response to Escherichia coli HSP70 (DnaK). Furthermore, intraperitoneal administration of DnaK resulted in a 4-fold increase in IL-12 in WT mice, which was absent in CD40 mice. This data demonstrates CD154-independent CD40 activation in polymicrobial sepsis and suggests that bacterial HSP70 is capable of stimulating CD40 in vitro and in vivo. PMID- 15545826 TI - Platelet-activating factor receptor-deficient mice show an unaltered clearance of nontypeable Haemophilus influenzae from their respiratory tract. AB - Platelet-activating factor (PAF), a glycerophospholipid with proinflammatory properties, exerts its biological effects by interacting with the PAF receptor (PAFR) expressed on many different cell types. The PAFR specifically binds phosphorylcholine, the biologically active component of PAF. However, phosphorylcholine is also a component of the cell wall of nontypeable Haemophilus influenzae (NTHi). In recently published in vitro experiments, the invasion of respiratory epithelial cells by NTHi was mediated by the PAFR. To determine the role of the PAFR in host defense against pneumonia induced by NTHi, PAFR deficient (PAFR-/-) and normal wild-type mice were intranasally inoculated with NTHi. The absence of a functional PAFR was associated with a normal innate immune response as indicated by similar bacterial counts, myeloperoxidase activity, and inflammation within the pulmonary compartment of PAFR-/- and wild-type mice. These data indicate that the PAFR does not interfere with the clearance of NTHi from the respiratory tract. PMID- 15545827 TI - Diminished ERK 1/2 and p38 MAPK phosphorylation in skeletal muscle during sepsis. AB - Sepsis induces weight loss and the loss of skeletal muscle proteins, in part through an inhibition of protein synthesis secondary to an inhibition of the key steps controlling mRNA translation in skeletal muscle. We have previously shown that sepsis decreases the phosphorylation of eIF4E. The present study examines the phosphorylation of Erk 1/2 MAPK and p38 MAPK in skeletal muscle of rats with a chronic (5-day) intra-abdominal septic abscess. Mnk1 catalyzes the phosphorylation of eIF4E, and Mnk1 is activated by phosphorylation via Erk1/2 MAPK and p38 MAPK. Sepsis resulted in a significant decrease in the steady-state phosphorylation of Erk 1/2 and p38 MAPKs compared with sterile inflammation. To examine the mediators responsible for decreased phosphorylation of Erk 1/2 and p38 MAPKs, rats were treated with TNF binding protein (TNFbp) or infused for 24 h with TNF. Treatment of septic rats with TNFbp resulted in an increase in the phosphorylation of both Erk 1/2 and p38 MAPKs in skeletal muscle. This was associated with enhanced phosphorylation of eIF4E. In contrast, constant intravenous infusion of TNF-alpha for 24 h resulted in a complete inhibition of p38 MAPK phosphorylation while Erk 1/2 MAPK phosphorylation was increased. The net effect was a modest increase in eIF4E phosphorylation. The results suggest altered regulation of Erk 1/2 and p38 MAPK signal translation pathways by endogenously produced TNF, or some compound dependent on TNF may modulate, in part, the phosphorylation state of eIF4E in skeletal muscle during sepsis. PMID- 15545828 TI - The peroxisome proliferator-activated receptor-gamma ligand 15-deoxyDelta12,14 prostaglandin J2 reduces the organ injury in hemorrhagic shock. AB - The cyclopentenone prostaglandin 15-deoxyDelta12,14PGJ2 (15d-PGJ2) exerts potent anti-inflammatory effects in vivo, which are in part caused by the activation of peroxisome proliferator-activated receptor-gamma (PPAR-gamma). Here we investigate the effects of 15d-PGJ2 on the multiple organ injury/dysfunction associated with severe hemorrhage and resuscitation. Male Wistar rats were subjected to hemorrhage (to lower mean arterial blood pressure to 45 mmHg) for 90 min and subsequently resuscitated with shed blood for 4 h. Rats were treated with either 15d-PGJ2 (0.3 mg/kg i.v.) or its vehicle (10% dimethyl sulfoxide) at 30 min before the hemorrhage. In some experiments, the selective PPAR-gamma antagonist GW9662 (1 mg/kg i.v.) or its vehicle (10% dimethyl sulfoxide) was given 45 min before the hemorrhage. Hemorrhage and resuscitation resulted in an increase in serum levels of (a) urea and creatinine and, hence renal dysfunction; alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and, hence, hepatic injury. The potent PPAR-gamma agonist 15d-PGJ2 abolished the renal dysfunction and largely reduced the liver injury caused by hemorrhagic shock. In addition, 15d-PGJ2 also attenuated the lung and intestinal injury (determined by histology) caused by hemorrhage and resuscitation. The specific PPAR-gamma antagonist GW9662 reduced the protective effects afforded by 15d-PGJ2. 15d-PGJ2 did not affect the delayed fall in blood pressure caused by hemorrhage and resuscitation. The mechanisms of the protective effect of this cyclopentenone prostaglandin are, at least in part, PPAR-gamma dependent, as the protection afforded by 15d-PGJ2 was reduced by the PPAR-gamma antagonist GW9662. We propose that 15d-PGJ2 or other ligands for PPAR-gamma may be useful in the therapy of the organ injury associated with hemorrhagic shock. PMID- 15545829 TI - Impact of the indigenous flora in animal models of shock and sepsis. AB - Septicemia is currently the 10th leading cause of death in the United States, and shock and trauma patients are the source of much of the morbidity and mortality associated with septicemia. There is substantial evidence that the composition of the indigenous flora plays an important role in modulating outcome variables in animal models of shock and sepsis. Germ-free animals that lack an indigenous flora are not as susceptible to shock as their conventionally reared counterparts. And, in conventionally reared animals, the composition of the intestinal flora can also modulate outcome in shock and sepsis. For example, certain bacterial species/strains disseminate from the intestinal tract more easily than others, antibiotic-induced alterations of the flora can modulate the incidence of systemic spread, and a certain threshold number of intestinal bacteria facilitates extraintestinal dissemination. The composition of the intestinal flora can also affect intestinal permeability, the production of inflammatory mediators, and the responses of immune cells in extraintestinal sites. And, there is evidence that prior exposure to endotoxin, via either the oral or systemic route, can influence outcome in animals challenged with parenteral endotoxin, a widely used model of endotoxin shock. The general composition of intestinal flora of experimental animals can be characterized with relative ease. This knowledge can aid data interpretation, either to help explain irreproducible or expected results or to verify that observed differences are likely related to the dependent variable studied rather than the composition of the indigenous flora. PMID- 15545830 TI - TSC for hemorrhagic shock: effects on cytokines and blood pressure. AB - Previous studies have shown that administering trans-sodium crocetinate (TSC) as a treatment of hemorrhagic shock leads to increased whole-body oxygen consumption and survival as well as protection of the liver and kidney. It has been suggested that TSC increases oxygen delivery by increasing the diffusivity of oxygen through plasma. However, as with any novel mechanism of action, there are always questions about whether the results could also be ascribed to other, previously described mechanisms of action. This study was designed to look at some aspects of that by examining the effect of different TSC dosing regimens on the blood pressure and the production of cytokines after hemorrhage because both responses have been reported with compounds that act via other mechanisms. In a constant pressure rat model of hemorrhagic shock, it was seen that a singe bolus injection of TSC results in an immediate but transient increase in the arterial blood pressure. This is similar to the effect reported previously for using 100% oxygen. It was also found that if the TSC injections were repeated periodically over an hour, a sustained increase in the blood pressure would occur. Because inflammatory cytokines have been implicated in mortality and tissue damage, it has been suggested that TSC may affect the production of cytokines. Thus, the effect of TSC on the production of TNF-alpha and IL-10 was also examined. The data show that treatment with TSC results in lower concentrations of TNF-alpha in the liver and spleen as well as lower concentrations of IL-10 in the spleen. Again, similar effects on other cytokines have been seen with 100% oxygen. These results support the hypothesis that the effects of TSC on hemorrhagic shock are mediated via an effect on oxygen. PMID- 15545831 TI - De novo synthesis of ubiquitin carboxyl-terminal hydrolase isozyme l1 in rostral ventrolateral medulla is crucial to survival during mevinphos intoxication. AB - Ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1) is a deubiquitinating enzyme that is responsible for making ubiquitin, which is required to target proteins for degradation by the ubiquitin-proteasome pathway in neurons, available. We investigated whether UCH-L1 plays a neuroprotective role at the rostral ventrolateral medulla (RVLM), the origin of sympathetic neurogenic vasomotor tone in the medulla oblongata where the organophosphate insecticide mevinphos (Mev) acts to elicit cardiovascular toxicity. In Sprague-Dawley rats maintained under propofol anesthesia, Mev (960 microg/kg, i.v.) induced a parallel and progressive augmentation in UCH-L1 or ubiquitin expression at the ventrolateral medulla during the course of Mev intoxication. The increase in UCH L1 level was significantly blunted on pretreatment with bilateral microinjection into the RVLM of a transcription inhibitor, actinomycin D (5 nmol), or a translation inhibitor, cycloheximide (20 nmol). Compared with aCSF or sense oligonucleotide (100 pmol) pretreatment, microinjection of an antisense uch-L1 oligonucleotide (100 pmol) bilaterally into the RVLM significantly increased mortality, reduced the duration of the "pro-life" phase, blunted the increase in ubiquitin expression in ventrolateral medulla, and augmented the induced hypotension in rats that received Mev. These findings suggest that de novo synthesis of UCH-L1, leading to an enhanced disassembly of ubiquitin-protein conjugates in the RVLM, is essential to maintenance of the "pro-life" phase of Mev intoxication via prevention of cardiovascular depression, leading to neuroprotection. PMID- 15545832 TI - Effects of sesame oil on oxidative stress after the onset of sepsis in rats. AB - The aim of this study was to investigate effects of sesame oil on oxidative stress after the onset of sepsis in rats. Effects of sesame oil on lipid peroxidation, superoxide anion, superoxide dismutase, catalase, glutathione, and nitrite after the onset of endotoxin intoxication were determined. To further examine the protective effect of sesame oil on sepsis, a mortality study was also conduced in cecal ligation and puncture-induced sepsis in rats. Sesame oil was given orally 6 h after endotoxin administration and cecal ligation and puncture, and parameters were then measured in another 6 h. Data demonstrated that a single dose of sesame oil reduced lipid peroxidation 6 h after endotoxin intoxication. Superoxide anion counts were decreased, glutathione levels were increased, and activities of superoxide dismutase and catalase, as well as nitrite levels, were not altered in lipopolysaccharide plus sesame oil-treated groups compared with lipopolysaccharide-treated groups. Furthermore, sesame oil given 6 h after cecal ligation and puncture significantly increased survival rate. Thus, we suggested that sesame oil could be used as a potent antioxidant to reduce oxidative stress after the onset of sepsis in rats. PMID- 15545833 TI - Vili in patients with sepsis: just fate or can we avoid it? PMID- 15545834 TI - Nitric oxide, bacteria, and host defense in sepsis: who needs what? PMID- 15545839 TI - Presentation of the 2004 Founders' Medal to J. Claude Bennett, MD. PMID- 15545840 TI - Founders' Medal 2004 acceptance address. PMID- 15545843 TI - Multiple Polymorphisms in the renin- angiotensin-aldosterone system (ACE, CYP11B2, AGTR1) and their contribution to hypertension in African Americans and Latinos in the multiethnic cohort. AB - OBJECTIVE: When compared with other U.S. populations, African Americans have excess hypertension. Genetic variants in elements of the renin-angiotensin aldosterone system (RAAS), namely the angiotensin-converting enzyme (ACE), aldosterone synthase (CYP11B2), and angiotensin II type 1 receptor (AGTR1) genes, have been associated with risk of hypertension in some populations. METHODS: We genotyped the D/I polymorphism in the ACE gene, the C(-344)T polymorphism in the CYP11B2 gene, and the C(-535)T polymorphism in the AGTR1 gene among African American and Latino members of the Multiethnic Cohort Study (MEC) to determine their association with hypertension. RESULTS: We observed no significant increase in the risk of hypertension for either African Americans or Latinos homozygous or heterozygous for the D allele of the ACE gene. Among African Americans we observed carriers of the (-344)T allele of CYP11B2 to be at increased risk of hypertension (versus CC genotype: TC genotype, OR = 1.66 [95% CI: 1.01-2.72]; TT genotype, OR = 1.74 [95% CI: 1.07-2.82]). There was also an increase in risk of hypertension associated with the AGTR1 T allele for African Americans (versus CC genotype: TC genotype, OR = 2.62 [95% CI: 1.46-4.72]; TT genotype, OR = 2.67 [95% CI: 1.51-4.74]). The associations observed with CYP11B2 and AGTR1 genotypes were not observed among Latinos. CONCLUSION: These data suggest that the (-535)T allele of AGTR1 and (-344)T allele of CYP11B2 may increase hypertension risk among African Americans but not among Latinos. Characterization of the linkage disequilibrium and haplotype patterns in the RAAS pathway genes will be crucial to understanding differences in hypertension susceptibility in these ethnic populations. PMID- 15545844 TI - A comparison of bedtime insulin glargine with bedtime neutral protamine hagedorn insulin in patients with type 2 diabetes: subgroup analysis of patients taking once-daily insulin in a multicenter, randomized, parallel group study. AB - BACKGROUND: Basal insulin is frequently administered once daily. This subgroup analysis of a multicenter, randomized, parallel study compared insulin glargine (Lantus Aventis Pharmaceuticals, Bridgewater, NJ) with neutral protamine Hagedorn (NPH) insulin in patients with type 2 diabetes, evaluating only patients treated previously with once-daily NPH insulin. METHODS: Patients received bedtime insulin glargine or NPH insulin, with preprandial regular insulin. One hundred patients (mean age, 57.9 years; mean glycohemoglobin, 8.4%; mean fasting blood glucose, 167 mg/dL) were treated for up to 28 weeks. RESULTS: Patients treated with insulin glargine (n = 52) and NPH insulin (n = 48) achieved similar reductions from baseline in glycohemoglobin (-0.41% versus -0.46%) and fasting blood glucose (-22 mg/dL versus -22 mg/dL) at week 28. The proportion of patients reaching target fasting blood glucose (<120 mg/dL) at 28 weeks was 34.2% with insulin glargine and 24.4% with NPH insulin. Similar proportions of patients achieved glycohemoglobin less than 7% and less than 8% in both groups. Baseline and week-28 mean daily doses of insulin glargine (27.3 IU versus 36.4 IU) were similar to NPH insulin doses (25.5 IU versus 30.2 IU). However, significantly fewer patients reported one or more episodes of hypoglycemia with insulin glargine (46.2%) versus NPH insulin (60.4%; P < 0.05). Significantly fewer patients also reported one or more symptomatic episodes confirmed by blood glucose less than 50 mg/dL with insulin glargine (17.3%) versus NPH insulin (31.3%; P < 0.005). CONCLUSION: Bedtime insulin glargine is as effective as bedtime NPH insulin in improving glycemic control, with significantly less hypoglycemia. PMID- 15545845 TI - Quadricuspid aortic valve: report of three cases. AB - Quadricuspid aortic valve (QAV) is a very rare congenital malformation. We have encountered three patients with QAV, of whom one patient may be the eldest reported patient with this particular anatomical abnormality. In another of our patients, there was aortic regurgitation, aortic stenosis, and healed infective endocarditis, with adhesion of the tips of the cusps. In all three patients, the cusps were all of equal size. Until now, there has been very little documented evidence about the anatomical variations in QAV or its relationship with infective endocarditis. From the available literature, we conclude that the anatomical variations in patients with QAV are similar to those in patients with quadricuspid pulmonary valve, and infective endocarditis may not be an uncommon complication. PMID- 15545846 TI - Metastatic pure small-cell carcinoma of prostate. AB - Extrapulmonary small-cell carcinoma arising in the prostate gland has been described in several case series and case reports. However, pure small-cell carcinoma of the prostate is rare, and there are only a few reports in literature describing the clinical features and management of this neoplasm. These tumors are highly aggressive and commonly manifest with visceral metastasis at the time of diagnosis. We report a case of metastatic pure prostatic small-cell carcinoma and an associated paraneoplastic polyneuropathy. PMID- 15545847 TI - Plasmapheresis as an adjuvant therapy for hypertriglyceridemia-induced pancreatitis. AB - Hypertriglyceridemia is an uncommon cause of pancreatitis. A serum triglyceride level of more then 1000 to 2000 mg/dL is an identifiable risk factor. Interestingly, serum pancreatic enzyme levels may be normal or only minimally elevated in such cases. The reduction of triglyceride level to below 1000 mg/dL effectively prevents further episodes of pancreatitis. The mainstay of treatment for the hypertriglyceridemia associated with pancreatitis includes dietary restriction of fat and administration of lipid-lowering agents. It is thought that within 24 to 48 hours of the onset of pancreatitis, in the majority of patients, triglyceride levels fall rapidly as a result of fasting status, as the absorption of chylomicrons to the blood is cut off. Experiences with plasmapheresis are limited. We report a case of hypertriglyceridemic necrotizing pancreatitis with mildly elevated amylase and lipase, treated successfully with plasmapheresis. PMID- 15545849 TI - Intrathoracic masses due to extramedullary hematopoiesis. AB - Extramedullary hematopoiesis often occurs in hemoglobinopathies, hemolytic anemias, leukemias, lymphomas, and myeloproliferative disorders. Liver, spleen, and lymph nodes are frequently involved. However, extramedullary hematopoiesis may also develop in other sites such as thymus, kidney, retroperitoneum, and paravertebral areas of the thorax. Extramedullary hematopoietic masses are often microscopic and asymptomatic, but sometimes they lead to tumor-like masses. We describe massive intrathoracic extramedullary hematopoiesis in a 41-year-old man with compound heterozygosis for beta-thalassemia and sickle cell anemia and functional asplenia. We also describe a 39-year-old man with beta-thalassemia intermedia, who was initially diagnosed as having tumor masses, but was later proved, by magnetic resonance imaging, to have extramedullary erythropoietic tissue. These observations provide further support to include extramedullary hematopoiesis among the differential diagnosis of tumor-like masses in patients with hematologic diseases. PMID- 15545848 TI - Thyrotoxicosis presenting as hypogonadism: a case of central hyperthyroidism. AB - Herein, we present a case of central thyrotoxicosis with well-documented serial therapeutic interventions. Thyroid-stimulating hormone (TSH)-secreting pituitary tumors represent a rare cause of hyperthyroidism. It is being diagnosed more frequently with the third-generation TSH assay. Many conditions can produce normal or elevated TSH levels in combination with elevated thyroid hormone levels. The differential diagnosis includes resistance to thyroid hormone (RTH, Refetoff's syndrome), assay interference from anti-T4/T3 and heterophile antibodies, elevated or altered binding proteins, drugs affecting peripheral metabolism, and noncompliance with thyroid replacement therapy. In contrast to RTH, our patient presented had high alpha-subunit-to-TSH molar ratio, failed TSH response to thyrotropin-releasing hormone stimulation, and a large pituitary mass. Normal or high TSH in the presence of elevated T4 or T3 is a fairly common clinical scenario with many etiologic possibilities. This TSH-producing adenoma represents an unusual initial clinical presentation, as hypogonadism appeared before features of thyrotoxicosis were appreciated. This case represents the most modern therapeutic approach to the management of this rare disease. Our patient has done well on octreotide with control of thyrotoxicosis and an additional 30% shrinkage of his tumor mass. PMID- 15545850 TI - Respiratory syncytial virus: G gene genotype and disease severity. AB - BACKGROUND: In a hospital-based study by Martinello (2002), specific G gene genotypes of respiratory syncytial virus subgroup A virus were associated with an increased severity of illness. AIM: We sought to confirm the association of G genotypes with disease severity in a population-based study. MATERIAL AND METHODS: Ninety-one type A respiratory syncytial viruses (identified in the 1999/2000 season by polymerase chain reaction and cell culture), collected in a German multicenter study (PRI.DE) were analyzed for G gene diversity (amino acids 1-165). Disease severity was classified according to World Health Organization criteria for pneumonia in outpatients and by a bronchiolitis score (Rodriguez, 1997) in inpatients. Multiple regression analysis was used to explain disease severity. RESULTS: Three clusters were identified (cluster 1, n = 35; cluster 2,n = 35; cluster 3, n = 21). Sixty-seven patients had severe disease. After controlling for other variables, illness severity was significantly greater for cluster 2 viruses (odds ratio, 7.0; 95% confidence interval, 1.6 49), compared with viruses in other clusters. Other known risk factors (male gender, age) were not associated with disease severity. Our cluster 2 is genetically distinct from the virulent genotype in Martinello's study. DISCUSSION: Previously reported associations between G genotypes and disease severity in hospitalized patients can be generalized across the spectrum of illnesses including outpatients. The association seems not to be linked to a specific G gene structure. Rather G gene diversity in combination with the susceptibility of the host cohort may form the basis of such associations. Because of the magnitude of the effect, the underlying mechanisms warrant further investigation. PMID- 15545851 TI - Infant pertussis: who was the source? AB - BACKGROUND: In the United States in the 1990s, the incidence of reported pertussis in adults, adolescents and infants increased; infants younger than 1 year of age had the highest reported incidence. METHODS: In 4 states with Enhanced Pertussis Surveillance, we examined the epidemiology of reported pertussis cases to determine the source of pertussis among infants. A source was defined as a person with an acute cough illness who had contact with the case infant 7-20 days before the infant's onset of cough. RESULTS: The average annual pertussis incidence per 100,000 infants younger than 1 year of age varied by state: 22.9 in Georgia; 42.1 in Illinois; 93.0 in Minnesota; and 35.8 in Massachusetts. Family members of 616 (80%) of 774 reported case-infants were interviewed; a source was identified for 264 (43%) of the 616 case-infants. Among the 264 case-infants, mothers were the source for 84 (32%) and another family member was the source for 113 (43%). Of the 219 source-persons with known age, 38 (17%) were age 0-4 years, 16 (7%) were age 5-9 years, 43 (20%) were age 10-19 years, 45 (21%) were age 20-29 years and 77 (35%) were age > or =30 years. CONCLUSIONS: The variation in reported pertussis incidence in the 4 states might have resulted from differences in awareness of pertussis among health care providers, diagnostic capacity and case classification. Among case-infants with an identifiable source, family members (at any age) were the main source of pertussis. Understanding the source of pertussis transmission to infants may provide new approaches to prevent pertussis in the most vulnerable infants. PMID- 15545852 TI - Risk of bacterial infection in previously healthy respiratory syncytial virus infected young children admitted to the intensive care unit. AB - OBJECTIVE: To evaluate the risk of bacterial infection and use of antibiotics in otherwise healthy children infected with respiratory syncytial virus (RSV) admitted to the intensive care unit (ICU). METHODS: Demographics, clinical information, interventions and outcomes were extracted from the charts of consecutive patients with laboratory-confirmed RSV infection at Children's Hospital, Boston from October 1990 through April 2002. Patients born at <36 weeks gestational age or with preexisting medical conditions were excluded. RESULTS: The median age of the 165 previously healthy infants infected with RSV was 42 days. Almost all patients received supplementary FiO2, and 63 (38.2%) patients required mechanical ventilator support. No patients died. The median length of stay was 3 days in the ICU and 7 days in the hospital. Most patients had bacterial cultures sent: 155 (93.9%), blood cultures; 121 (73.3%), urine cultures; and 85 (51.5%) cerebrospinal fluid cultures. Only 1 blood culture was positive, and 1 potential urinary tract infection was identified in a patient with a negative urinalysis. All intubated patients and 80.4% of nonintubated patients received antibiotic therapy. CONCLUSIONS: In otherwise healthy infants admitted to the ICU with RSV infection, bacteremia, urinary tract infection and meningitis are uncommon. Although bacterial pneumonia in this cohort may be more prevalent, overdiagnosis is common. PMID- 15545853 TI - Rhinovirus-associated wheezing in infancy: comparison with respiratory syncytial virus bronchiolitis. AB - BACKGROUND: There is increasing evidence that rhinoviruses (RV) are able to cause lower airway infections and to induce wheezing in young children. There are few data on the clinical characteristics of RV infections in infants. OBJECTIVE: The aim of the study was to compare clinical characteristics of infantile RV infection associated with wheezing and respiratory syncytial virus (RSV) bronchiolitis. MATERIAL AND METHODS: During a 22-month study period in 1992-1993, 100 children younger than 24 months old were hospitalized with respiratory tract infection-associated wheezing. Viral etiology was originally assessed by antibody and antigen assays. Etiologic studies were later supplemented by polymerase chain reaction for RVs (in 2000) and for RSV (in 2002), studied in frozen respiratory samples. There were 81 children with adequate determinations for both RVs and RSV. Twenty-six children had RV and 24 had RSV infection, and these 50 cases form the material of the present study. Atopic dermatitis, oxygen saturation, respiratory rates and clinical scores based on wheezing and retractions and total serum IgE concentrations and blood eosinophil counts were studied in all cases on admission. RESULTS: The children with RV infection, compared with RSV patients, were older (median, 13 versus 5 months), presented more often with atopic dermatitis (odds ratio, 16.7; 95% confidence interval, 2.22-100) and blood eosinophilia (odds ratio, 2.22; 95% confidence interval, 1.04-50). The groups did not differ from each other with regard to total serum IgE. Oxygen saturation values were lower in children with RSV infection. There were no significant differences in respiratory rates or scores combining wheezing and retractions. CONCLUSION: RV-associated wheezing and RSV bronchiolitis, although having rather similar clinical characteristics, differ significantly with regard to age, presence of atopic dermatitis and eosinophilia during infection. PMID- 15545854 TI - Viral etiology of acute respiratory tract infections in children presenting to hospital: role of polymerase chain reaction and demonstration of multiple infections. AB - BACKGROUND: Viral lower respiratory tract infections are a leading cause of hospitalization for young children. METHODS: We used polymerase chain reaction (PCR) and conventional methods of cell culture and antigen detection to establish the viral etiology of acute respiratory tract infections in 75 hospitalized children. RESULTS: One or more viral pathogens were detected in 65 (87%) children, with respiratory syncytial virus being the most commonly identified virus (36 children). Other viruses identified included influenza virus types A and B, parainfluenzavirus type 3, adenovirus, enterovirus, rhinovirus, coronavirus and human metapneumovirus. PCR increased the diagnostic yield significantly compared with antigen detection and culture, with 39 (21%) diagnoses identified by this method. Multiple infections were identified in 20 (27%) children. CONCLUSIONS: PCR-based methodologies offer increased sensitivity for the detection of most respiratory viruses in young children. The inclusion of PCR into diagnostic testing strategies is needed to broaden our understanding of the natural ecology of respiratory viruses and the significance of multiple infections. PMID- 15545855 TI - Immunogenicity and safety of the eleven valent pneumococcal polysaccharide protein D conjugate vaccine in infants. AB - BACKGROUND: Development is ongoing to increase the serotype coverage of pneumococcal conjugate vaccines. We report here the immunogenicity and safety of a new 11-valent pneumococcal conjugate vaccine (Pn-PD) in infants. METHODS: In a randomized, single blind study, 154 Finnish infants received 1 of 3 regimens: 4 doses of Pn-PD at 2, 4, 6 and 12-15 months; 3 doses of the Pn-PD at 2, 4 and 6 months and 1 dose of 23-valent polysaccharide vaccine (PncPS) at 12-15 months; or 3 doses of the hepatitis B vaccine at 2, 4 and 6 months and Pn-PD at 12-15 months. Serum IgG antibodies to vaccine serotypes 1, 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F were measured with an enzyme immunoassay at the ages of 2, 7 and 12-15 months and at 4 or 28 days after the last vaccination. Local and systemic reactions were recorded by parents during 8 days after each dose. Serious adverse reactions were recorded during the entire study period. RESULTS: There was a significant increase in the IgG concentrations to vaccine serotypes after 3 doses of Pn-PD. Antibody concentrations after the primary series varied between 1.26 and 4.92 microg/ml depending on the serotype and study group. PncPS vaccine induced a better booster response than the Pn-PD, measured at 28 days after the fourth dose. IgG concentrations after the Pn-PD booster ranged between 1.60 and 9.63 microg/ml and after the PncPS booster between 4.24 and 40.54 microg/ml, depending on the serotype. The antibody concentrations after the first dose of Pn PD administered at 12-15 months increased significantly but were lower than after the fourth dose at the same age. No significant antibody increase was measured 4 days after the vaccinations at 12-15 months. The safety profile of the vaccine was acceptable. CONCLUSIONS: The Pn-PD we tested was immunogenic and safe in infants. PMID- 15545856 TI - Seven valent pneumococcal conjugate vaccine immunization in two Boston communities: changes in serotypes and antimicrobial susceptibility among Streptococcus pneumoniae isolates. AB - BACKGROUND: Seven valent pneumococcal conjugate vaccine (PCV7) was licensed and introduced in 2000 for universal administration of children younger than 2 years of age and for selective immunization of children 2-5 years of age. SPECIFIC AIMS: To identify changes in colonization and antimicrobial susceptibility among Streptococcus pneumoniae organisms after introduction of PCV7. METHODS: Infants and children ages 2-24 months were enrolled in surveillance study of nasopharyngeal carriage of S. pneumoniae. Nasopharyngeal cultures for S. pneumoniae were performed at all well child visits and illness visits of children with acute otitis media. S. pneumoniae organisms were serotyped, and antimicrobial susceptibilities to penicillin, amoxicillin, trimethoprim sulfamethoxazole and azithromycin were performed. RESULTS: During the 3-year period (October 2000 through September 2003), nasopharyngeal colonization with vaccine serotypes declined from 22% to 2%, and nonvaccine serotypes increased from 7% to 16%. Rates of antibiotic resistance of S. pneumoniae isolates to penicillin, amoxicillin, azithromycin and trimethoprim-sulfamethoxazole were 29.3, 2.2, 26.5 and 28.1%, respectively. CONCLUSIONS: PCV7 immunization produces a marked decline in vaccine serotypes carried in the nasopharynx of young children, with a coincident rise in the prevalence of nonvaccine serotypes. Important shifts in antimicrobial susceptibility have not been observed to date. PMID- 15545857 TI - Serotype-specific pneumococcal antibodies in breast milk of Gambian women immunized with a pneumococcal polysaccharide vaccine during pregnancy. AB - BACKGROUND: In breast-feeding populations, immunization during pregnancy with pneumococcal polysaccharide offers a potentially useful approach to preventing pneumococcal disease in young infants. METHODS: Breast milk samples were collected at 0, 2, 4 and 6 months after delivery from Gambian women vaccinated during pregnancy (24-32 weeks gestation) with Pneumovax II (n = 56) or Mengivax A&C (n = 57). Specimens were examined for secretory immunoglobulin A (s-IgA) concentration, subclass distribution and avidity specific to pneumococcal serotypes 4, 6B, 14, 19F and 23F and the antigen mixture in Pneumovax II by enzyme-linked immunosorbent assay. Colostral s-IgA and IgG concentrations in paired maternal sera were compared. RESULTS: Colostral s-IgA concentrations specific to all pneumococcal polysaccharide antigens investigated were significantly higher (P < 0.05) among Pneumovax II vaccinees. Titers specific to serotypes 4, 6B and 14 and the vaccine formula remained significantly higher during 6 months, and those for 19F were higher during 4 months. Significantly higher concentrations of vaccine antigen-specific s-IgA antibody were sustained for 6 months after delivery (P = 0.011). Comparison of colostral s-IgA and IgG in serum revealed a significant correlation only among Mengivax A&C vaccinees for pneumococcal polysaccharide 23F (rs= 0.68; P < or = 0.0001). Vaccination elicited trends toward increased s-IgA2, reaching significance for serotype 14 and the vaccine formula. Immunization elicited significantly higher s-IgA avidities specific to all pneumococcal polysaccharide antigens studied during 6 months. CONCLUSIONS: The public health value of immunization during pregnancy with pneumococcal polysaccharide vaccine in breast-feeding populations warrants further evaluation, particularly in populations with a high incidence of pneumococcal disease in early infancy. PMID- 15545858 TI - Otoscopic and tympanometric findings in acute otitis media yielding dry tap at tympanocentesis. AB - BACKGROUND: The value of tympanometry in detection of middle ear effusion (MEE) has been widely studied in otitis media with effusion. There has been no direct comparison of tympanometric and tympanocentesis (TAP) findings in acute otitis media (AOM). We compared otoscopic, tympanometric and TAP findings in AOM including cases of AOM without effusion. METHODS: In a study of AOM treatment of 90 children, a tympanogram was obtained, and TAP was performed before and after 5 days of treatment. Subjects were followed with otoscopy and tympanometry every 2 weeks for 3 months or until the MEE cleared. RESULTS: In 130 AOM ears, otoscopic, tympanometric and TAP findings were available; MEE was obtained from 110 ears. Of 20 ears with a dry tap, 18 had abnormal tympanogram and otoscopic findings. With TAP findings as the standard, sensitivity and positive predictive value of type B tympanogram were 97 and 87%, respectively. Of 18 AOM ears yielding dry tap, 2 yielded MEE 5 days later, and 8 continued with evidence of MEE 5 days-12 weeks later. Five subjects with 8 AOM ears yielding dry tap were lost to follow-up; all had short duration of clinical symptoms. CONCLUSIONS: Although otoscopic and tympanometric findings suggested the presence of MEE in AOM, MEE was unobtainable by TAP in 14% of cases. Dry tap cases likely represent early AOM before accumulation of detectable MEE. However, technical difficulty in obtaining small amounts of or highly viscous MEE could not be excluded. Sensitivity and positive predictive value of abnormal tympanograms in detection of MEE in AOM cases are comparable with those in otitis media with effusion. PMID- 15545859 TI - The role of psychosocial and family factors in adherence to antiretroviral treatment in human immunodeficiency virus-infected children. AB - BACKGROUND: The toxicity and complexity of antiretroviral therapy (ART) regimens are substantial challenges in the context of patients' lives. This study examines child psychosocial and caregiver/family factors influencing adherence to ART in perinatally human immunodeficiency virus (HIV)-infected children. METHODS: Seventy-five children (ages 3-13 years) prescribed ART, and their primary caregivers were recruited from 2 urban pediatric HIV programs. A battery of psychologic assessments and self-report adherence data were collected from all caregivers and 48 children who were > or =7 years old. RESULTS: Forty percent of caregivers and 56% of children reported missed doses of medication in the past month. Families in which the caregiver or child reported missed doses (nonadherent) were compared with families who reported no missed doses (adherent). In univariate analyses, nonadherence was significantly associated with older child age (P < 0.05), worse parent-child communication (P < 0.017), higher caregiver stress (P < 0.002), lower caregiver quality of life (P < 0.003) and worse caregiver cognitive functioning (P = 0.033), and of borderline significance in its association with increased (1) child responsibility for medications (P < 0.07), (2) HIV disclosure to the child (P < 0.07) and (3) child stress (P < 0.08) In logistic regressions controlling for age, caregiver/family factors were the most strongly associated with nonadherence, including worse parent-child communication (P < 0.03), higher caregiver stress (P < 0.01), less disclosure to others (P < 0.05) and quality of life (P < 0.01). CONCLUSIONS: Our data suggest that efforts to improve children's adherence to complex antiretroviral regimens requires addressing developmental, psychosocial and family factors. PMID- 15545860 TI - Vertical dengue infection: case reports and review. AB - Two vertical dengue infection cases are presented, and 15 others are reviewed. Twelve mothers had fever 0-9 (median, 2) days antepartum. The fevers of 17 neonates occurred at 1-11 (median, 4) days of life and lasted for 1-5 (median, 3) days. Neonatal thrombocytopenia was detected at 1-11 (median, 6) days of life and lasted for 3-18 (median, 6) days; the lowest platelet counts were 5-75 x 10(3) (median, 19 x 10 (3))/mm3. One neonate died. PMID- 15545861 TI - Coronaviruses: from common colds to severe acute respiratory syndrome. PMID- 15545862 TI - Controversies in palivizumab use. PMID- 15545863 TI - Live attenuated influenza vaccine induces cross-reactive antibody responses in children against an a/Fujian/411/2002-like H3N2 antigenic variant strain. AB - Serum antibody titers against the A/Panama/2007/99(H3N2) and A/Fujian/411/2002(H3N2)-like viruses were determined in children 6-35 months of age who received either 1 dose of the inactivated influenza vaccine or the live attenuated influenza vaccine containing the A/Panama strain. Results indicated that the live vaccine induced higher antibody responses than the inactivated vaccine against the A/Panama and A/Fujian-like viruses. PMID- 15545864 TI - Detection of Helicobacter pylori in the stools of newborn infants. AB - To investigate the transmission route of Helicobacter pylori, stool specimens of 50 infants 3 days old were studied by a stool antigen immunoassay and polymerase chain reaction (PCR). By PCR, H. pylori DNA was detected in 15 neonates (30%). The stool antigen test was positive in only 1 neonate with a positive PCR. The detection rate of H. pylori DNA in neonates was significantly higher for those with mothers having urines positive for anti-H. pylori IgG antibody (60%) than for those mothers with negative urines (17%) (P < 0.01). A follow-up study was done 24 months later on 8 of the 15 infants with positive PCR results, including the infant with a positive stool antigen test. All infants were negative by both PCR and stool antigen test. PMID- 15545865 TI - Alternative, age- and viral load-related routes of nelfinavir resistance in human immunodeficiency virus type 1-infected children receiving highly active antiretroviral therapy. AB - To assess prevalence of nelfinavir resistance mutations in children receiving highly active antiretroviral therapy, sequencing of protease gene from plasma of 53 human immunodeficiency virus-infected children was performed. The prevalence of L90M was similar to that of D30N. There was a significant correlation with a higher viral load and lower age and the occurrence of L90M. These findings suggest differential molecular age- and viral load-related routes for nelfinavir resistance. PMID- 15545866 TI - Fusarium solani endocarditis successfully treated with liposomal amphotericin B and voriconazole. AB - Fungal infections caused by Fusarium in the immunocompromised host are highly resistant to all antifungal agents. Fusarium endocarditis is a rare and usually fatal disease. We report an immunocompromised child who survived Fusarium solani endocarditis despite the in vitro resistance of the organism to all available antifungal agents. PMID- 15545867 TI - A cluster of early neonatal sepsis and pneumonia caused by nontypable Haemophilus influenzae. AB - Nine premature infants developed early onset sepsis and/or pneumonia with Haemophilus influenzae during a period of 53 months (January 2000 -May 2004). Their respiratory problems were pneumonia-like rather than classic respiratory distress syndrome. In 8 of the cases, the pathogen was a beta-lactamase-negative, nontypable H. influenzae. In the remaining case, the Haemophilus identified was type d. Before January 2000, no case of beta-lactamase-negative, nontypable H. influenzae sepsis or pneumonia had been recorded. PMID- 15545868 TI - Pasteurella multocida meningitis and cervical spine osteomyelitis in a neonate. AB - A 20-day-old male infant presented with fever, decreased alertness and quadriparesis as a result of Pasteurella multocida meningitis and C1-2 vertebral osteomyelitis. Although his household contained 2 pet cats, there was no history of bites, scratches or licks. We speculate that colonization of the nasopharynx was followed by contiguous spread to the retropharyngeal soft tissue, cervical vertebrae and meninges. PMID- 15545869 TI - Detection of subgroup B respiratory syncytial virus in the cerebrospinal fluid of a patient with respiratory syncytial virus pneumonia. AB - We report detection of subgroup B respiratory syncytial virus in cerebrospinal fluid from a 4-month-old infant, using a 1-step reverse transcription-polymerase chain reaction assay. The infant was diagnosed with bilateral pneumonia and had a febrile seizure. The patient recovered uneventfully after 9 days in the hospital. PMID- 15545870 TI - Two enigmatic cases of meningitis. PMID- 15545871 TI - Two distinct patterns of pleural effusions caused by Mycoplasma pneumoniae infection. PMID- 15545873 TI - Predictive model for childhood meningitis. PMID- 15545874 TI - Prolonged treatment with albendazole for massive trichuriasis infection. PMID- 15545876 TI - Therapy for Mycobacterium bovis. PMID- 15545877 TI - Diagnosis of orthopedic infection in clinical practice using Tc-99m sulesomab (antigranulocyte monoclonal antibody fragment Fab'2). AB - OBJECTIVE: The aim of the study was to analyze the use of Tc-99m-labeled antigranulocyte monoclonal antibody fragment Fab'2 (SMab, Leukoscan) in the diagnosis of acute osteomyelitis and articular prosthesis infection. One hundred seventeen patients (118 studies) have been included, 92 women and 25 men. Thirty seven had a suspicion of osteomyelitis and 81 with suspicion of infection of a hip (35 patients) or a knee (45 patients) prosthesis. The mean age was 64 years. A 3-phase bone scan and a SMab scan were obtained in all patients. Antigranulocyte scans were obtained within 3 and 8 hours postinjection of 20 mCi (740 MBq) of Tc-99m-labeled antibody. Diagnosis of infection was based on bone biopsy in 8 cases and on at least 2 positive cultures in the remaining cases. Infection was ruled out with negative cultures or 1-year patient follow up with negative C-reactive protein and normal erythrocyte sedimentation rate. RESULTS: Sensitivity, specificity, accuracy, and negative predictive value of Tc 99m-SMab scintigraphy were: 75%, 95%, 87%, and 92% for long bone osteomyelitis and 80%, 89%, 87%, and 91% for articular prosthesis infection, respectively. Positive predictive values were 83% and 63%, respectively. CONCLUSION: Tc-99m antigranulocyte monoclonal antibody fragment Fab'2 scintigraphy seems to be a useful method in the diagnosis of osteoarticular infection. PMID- 15545878 TI - F-18 FDG PET/CT in acute respiratory distress syndrome: a case report. AB - F-18 FDG PET/CT has become a useful technique in the evaluation of pulmonary lesions. We present a case of markedly increased and diffuse pulmonary F-18 FDG activity in a patient with acute respiratory distress syndrome (ARDS). High rates of glucose utilization by the inflammatory cells involved in the pathogenesis of ARDS might explain the increased pulmonary F-18 FDG uptake we observed. In the proper clinical setting, ARDS should be considered in the differential diagnosis of patients with diffusely increased F-18 FDG activity in the lungs. PMID- 15545879 TI - FDG PET in epithelioid hemangioendothelioma. AB - Epithelioid hemangioendothelioma (EH) is an uncommon tumor of endothelial origin. It can develop in any tissue and can be multicentric or metastatic. The usual course is a slow progression. Imaging techniques are generally useful in determining the extent of the disease. A case of EH involving bone marrow and mediastinum is described. We discuss the use of FDG PET scanning in EH, showing its use in detecting bone marrow involvement and determining the extent of the disease. PMID- 15545880 TI - Liver and vertebral uptake of Tc-99m macroaggregated albumin (MAA). AB - An unusual phenomenon during lung scintigraphy is presented. Besides visualization of the lungs, accumulation of Tc-99m macroaggregated albumin (MAA) was seen in a small part of the liver and in and around several thoracic vertebrae. Contrast-enhanced radiographic computed tomography revealed extensive collateral pathways, which were caused by a partially obstructed superior vena cava. Shunting of systemic venous blood flow through chest wall veins to the portal system was responsible for accumulation of MAA in the liver. Retrograde blood flow through dilated thoracic vertebral veins resulted in visualization of the bone marrow. PMID- 15545881 TI - Post I-131 therapy scanning in patients with thyroid carcinoma metastases: an unnecessary cost or a relevant contribution? AB - RATIONALE: In patients with elevated serum thyroglobulin (Tg) and a negative whole-body scan (WBS), posttherapy scanning can reveal metastases in many cases and is therefore recommended. In contrast, the routine use of posttherapy scanning is questioned. MATERIALS AND METHODS: One hundred six patients with differentiated thyroid carcinoma had pre- and posttherapy scanning, with metastatic uptake on the diagnostic scan and were divided into 2 groups: 60 patients assessed on first ablation after thyroidectomy and 46 patients with tumor persistence or recurrence after initial ablative therapy. Data modifying the disease stage or treatment were considered to be clinically relevant. RESULTS: Posttherapy scanning on first ablation changed the disease stage in 8.3% of the patients and therapeutic approach in another 15%, and provided clinically relevant information for 26% of patients with 1 previous ablation. Even when excluding cases whose lesions were known by the time of the first postablative scan, the therapeutic approach was influenced by posttherapy scanning in 15.6% of the patients. Only 4 of 211 metastases detected on pretherapy WBS did not appear on postablative scans. CONCLUSIONS: Posttherapy scanning provides important information, even in patients whose pretherapy WBS is positive for metastases, with this approach being useful both during the first ablation and subsequent treatment. PMID- 15545882 TI - Dual Tc-99m sestamibi and Gallium-67 SPECT localize a myocardial abscess around a bioprosthetic aortic valve. PMID- 15545883 TI - Tc-99m leukocyte imaging in acute graft versus host disease. PMID- 15545884 TI - Rhabdomyolysis resulting from interaction of simvastatin and clarithromycin demonstrated by Tc-99m MDP scintigraphy. PMID- 15545885 TI - Intense bilateral pectoralis major muscle activity on Tc-99m MDP scintigraphy: the push-up sign. PMID- 15545886 TI - FDG uptake in intercostal muscles is an indicator of severe respiratory disease. PMID- 15545887 TI - F-18 PET soft tissue uptake in seatbelt injury. PMID- 15545888 TI - Melanoma metastasis to the testis demonstrated with FDG PET/CT. PMID- 15545889 TI - Fibula stress fracture and confounding myositis ossificans. PMID- 15545890 TI - A severe case of enthesopathy mimicking intertrochanteric fractures on Tc-99m MDP scan. PMID- 15545891 TI - Combined Tc-99m MDP and Tc-99m labeled RBC imaging in a case of Maffucci's syndrome. PMID- 15545892 TI - Tumoral calcinosis appearances on skeletal scintigraphy. PMID- 15545893 TI - Meckel's diverticulitis: diagnosis with computed tomography and Tc-99m pertechnetate scintigraphy. PMID- 15545894 TI - FDG PET and gallium scintigraphy for diagnosis of an advanced jejunal adenocarcinoma with distant metastases. PMID- 15545895 TI - Epithelioid hemangioendothelioma of the liver with F-18 FDG PET imaging. PMID- 15545896 TI - Unusual ventilation-perfusion scintigraphy in adult respiratory distress syndrome secondary to acute lupus myocarditis. PMID- 15545897 TI - Localization of F-18 FDG in Nocardia lung infection. PMID- 15545898 TI - Pleural cerebrospinal fluid input and output kinetics dynamically demonstrated by In-111 DTPA myelography in a patient with pleural cerebrospinal fluid fistulae. PMID- 15545899 TI - Methionine and glucose metabolism of central neurocytoma: a PET study. PMID- 15545900 TI - Bilateral axillary sentinel nodes from breast carcinoma. PMID- 15545901 TI - Ectopic acromegaly: localization of the pituitary growth hormone-releasing hormone producing tumor by In-111 pentetreotide scintigraphy and report of two cases. PMID- 15545902 TI - Usefulness of Tc-99m RBC SPECT/MRI fusion imaging in small suspected hepatic hemangiomas. PMID- 15545903 TI - Incidental situs inversus visualized with FDG PET/CT. PMID- 15545904 TI - Uptake of FDG in the area of a recently implanted bioprosthetic mitral valve. PMID- 15545905 TI - Focal thyroid uptake on bone scan due to thyroid biopsy. PMID- 15545906 TI - Supernumerary left kidney. PMID- 15545907 TI - Heart and soft-tissue uptake of Tc-99m MDP in a hemodialysis patient. PMID- 15545908 TI - Abnormal bone scan in an adult with osteopoikilosis. PMID- 15545909 TI - F-18 FDG PET and Ga-67 scintigraphy in a case of fever of unknown origin with underlying cutaneous diffuse large B-Cell lymphoma. PMID- 15545910 TI - Technetium-99m-labeled RBC bleeding scan in a patient with Osler-Weber-Rendu Disease. PMID- 15545911 TI - Current readings in nuclear medicine. PMID- 15545912 TI - Targeting anti-transferrin receptor antibody (OX26) and OX26-conjugated liposomes to brain capillary endothelial cells using in situ perfusion. AB - Brain capillary endothelial cells (BCECs) express transferrin receptors. The uptake of a potential drug vector (OX26, or anti-transferrin receptor antibody IgG2a) conjugated to polyethyleneglycol-coated liposomes by BCECs was studied using in situ perfusion in 18-day-old rats in which the uptake of OX26 is almost twice as high as in the adult rat. Using radio-labeling, the uptake of OX26 by BCECs after 15-minute perfusion was approximately 16 times higher than that of nonimmune IgG2a (Ni-IgG2a). OX26 and OX26-conjugated liposomes selectively distributed to BCECs, leaving choroid plexus epithelium, neurons, and glia unlabeled. Ni-IgG2a and unconjugated liposomes did not reveal any labeling of BCECs. The labeling of BCECs by OX26 was profoundly higher than that of transferrin. Perfusion with albumin for 15 minutes did not reveal any labeling of neurons or glia, thus confirming the integrity of the blood-brain barrier. The failure to label neurons and glia shows that OX26 and OX26-conjugated liposomes did not pass through BCECs. The expression of transferrin receptors by endothelial cells selective to the brain qualifies OX26 as a candidate for blood to-endothelium transport. A specifically designed formulation of liposomes may allow for their degradation within BCECs, leading to subsequent transport of liposomal cargo further into the brain. PMID- 15545913 TI - Postischemic intraventricular administration of FGF-2 expressing adenoviral vectors improves neurologic outcome and reduces infarct volume after transient focal cerebral ischemia in rats. AB - Fibroblast growth factor (FGF)-2 is a potent neurotrophic and angiogenic peptide. To examine possible protective effects of FGF-2 gene expression against transient focal cerebral ischemia in rats, a replication defective, recombinant adenovirus vector expressing FGF-2, was injected intraventricularly 2 hours after middle cerebral artery occlusion (MCAO). The treatment group showed significant recovery compared with the vehicle-treated groups in terms of serial neurologic severity scores over the 35 days after MCAO. Further, 2,3,5-triphenyltetrazolium chloride staining showed that FGF-2 gene transfer decreased infarct volume by 44% as compared with that in the vehicle-treated groups at 2 days after MCAO. The same tendency of gene transfer effects on infarct volume was confirmed at 35 days after MCAO with hematoxylin/eosin staining. Enzyme-linked immunosorbent assay revealed that FGF-2 concentration was increased significantly at 2 days after MCAO, not only in cerebrospinal fluid but also in cerebral substance in the lesioned and treated animals. These results suggested that FGF-2 gene transfer using these adenoviral vectors might be a useful modality for the treatment of occlusive cerebrovascular disease even after the onset of stroke. PMID- 15545914 TI - Human C-reactive protein increases cerebral infarct size after middle cerebral artery occlusion in adult rats. AB - Human C-reactive protein (CRP), the classic acute phase plasma protein, increases in concentration after myocardial infarction and stroke. Human CRP binds to ligands exposed in damaged tissue and can then activate complement and its proinflammatory functions. In contrast, rat CRP, which binds to similar ligands, does not activate complement. In the present study, systemic complement depletion with cobra venom factor in adult rats subjected to middle cerebral artery occlusion did not affect cerebral infarct size, indicating that circulating complement does not contribute to injury in this model. However, we have previously reported that administration of human CRP to rats undergoing coronary artery ligation caused a marked increase in size of the resulting myocardial infarction, associated with codeposition of human CRP and rat complement in the infarcts. In the present study, we show that adult rats subjected to middle cerebral artery occlusion and then treated with human CRP similarly developed significantly larger cerebral infarcts compared with control subjects receiving human serum albumin. Human CRP can thus contribute to ischemic tissue damage in the brain as well as in the heart, and inhibition of CRP binding may therefore be a promising target for tissue protective acute therapeutic intervention in stroke as well as in myocardial infarction. PMID- 15545916 TI - Valproate is neuroprotective against malonate toxicity in rat striatum: an association with augmentation of high-affinity glutamate uptake. AB - The antiepileptic drug valproate (VPA) may be neuroprotective. We treated rats with VPA for 14 days (300 mg/kg twice daily) before intrastriatal injection of 1.5 micromol (1 M) of the succinate dehydrogenase inhibitor malonate. VPA-treated animals developed smaller lesions than control animals: 10 +/- 2 mm(3) versus 26 +/- 8 mm(3) (means +/- SD; P = 10(-4). Injection of NaCl that was equiosmolar with 1 M malonate caused lesions of only 1.2 +/- 0.4 mm(3) in control animals, whereas physiologic saline produced no lesion. VPA pretreatment reduced the malonate-induced extracellular accumulation of glutamate. This effect paralleled an increase in the striatal level of the glutamate transporter GLT, which augmented high-affinity glutamate uptake by 25%, as determined from the uptake of [(3)H] glutamate into striatal proteoliposomes. Malonate caused a 76% reduction in striatal adenosine triphosphate (ATP) content, but the glial, ATP-dependent formation of glutamine from radiolabeled glucose or glutamate was intact, indicating that glial ATP production supported uptake of glutamate. Striatal levels of HSP-70 and fos were reduced, and the levels of bcl-2 and phosphorylated extracellular signal-regulated kinase remained unaffected, but histone acetylation was increased by VPA treatment. The results suggest that augmentation of glutamate uptake may contribute importantly to VPA-mediated neuroprotection in striatum. PMID- 15545915 TI - Protein ubiquitination in postsynaptic densities after transient cerebral ischemia. AB - The mechanisms underlying neurologic deficits and delayed neuronal death after ischemia are not fully understood. In the present study, we report that transient cerebral ischemia induces accumulation of ubiquitinated proteins (ubi-proteins) in postsynaptic densities (PSDs). By immunoelectron microscopy, we demonstrated that ubi-proteins were highly accumulated in PSD structures after ischemia. On Western blots, ubi-proteins were markedly increased in purified PSDs at 30 minutes of reperfusion, and the increase persisted until cell death in the CA1 region after ischemia. In the resistant DG area, however, the changes were transient and significantly less pronounced. Deposition of ubi-proteins in PSDs after ischemia correlates well with PSD structural damage in the CA1 region as viewed by electron microscopy. These results suggest that the ubiquitin proteasome system fails to repair and remove damaged proteins in PSDs. The changes may demolish synaptic neurotransmission, contribute to neurologic deficits, and eventually lead to delayed neuronal death after transient cerebral ischemia. PMID- 15545918 TI - Empiricism and rationalism: two paths toward the same goal. PMID- 15545917 TI - The role of hypotheses in current research, illustrated by hypotheses on the possible role of astrocytes in energy metabolism and cerebral blood flow: from Newton to now. PMID- 15545919 TI - The astrocyte-neuron lactate shuttle: a challenge of a challenge. PMID- 15545920 TI - Does the acute diffusion-weighted imaging lesion represent penumbra as well as core? A combined quantitative PET/MRI voxel-based study. AB - In acute ischemic stroke, the diffusion-weighted imaging (DWI) lesion is widely held to represent the core of irreversible damage and is therefore crucial in selecting patients for thrombolysis. However, recent research suggests it may also represent penumbra. An illustrative patient was imaged 7 hours after stroke onset with back-to-back 3T diffusion tensor imaging and quantitative positron emission tomography, which showed a DWI lesion and misery perfusion, respectively. Using previously validated voxel-based probabilistic CBF, CMRO2, and Oxygen Extraction Fraction (OEF) thresholds, the authors show that the DWI lesion contained not only core but also substantial proportions of penumbra. Also, severe apparent diffusion coefficient reductions were present within the potentially salvageable penumbra as well as in the core. These findings have potential implications regarding treatment decisions. PMID- 15545921 TI - Degeneration of the basalocortical pathway from the cortex induces a functional increase in galaninergic markers in the nucleus basalis magnocellularis of the rat. AB - The present work aimed 1) to evaluate whether an increase in galanin or galanin receptors could be induced in the nucleus basalis magnocellularis (nbm) by degeneration of the basalocortical neurons from the cortex and 2) to analyze the consequences of such an increase on cortical activity. First, a mild ischemic insult to the frontoparietal cortex was performed to induce the degeneration of the basalocortical system; galanin immunoreactivity, galanin binding sites, and cholinergic muscarinic receptors were quantified through immunocytochemistry and autoradiography. Second, galanin infusions in the nbm were undertaken to mimic a local increase of the galaninergic innervation; cortical acetylcholine release, cerebral glucose use, and cerebral blood flow were then measured as indices of cortical activity. As a result of the cortical ischemic lesion, the postsynaptic M1 and presynaptic M2 muscarinic receptors were found to be reduced in the altered cortex. In contrast, galaninergic binding capacity and fiber density were found to be increased in the ipsilateral nbm in parallel with a local decrease in the cholinergic markers such as the muscarinic M1 receptor density. Galanin infusion into the nbm inhibited the cortical acetylcholine release and cerebral blood flow increases elicited by the activation of the cholinergic basalocortical system but failed to affect acetylcholine release, cerebral blood flow, and cerebral glucose use when injected alone in the nbm. These results demonstrate that degeneration of the basalocortical system from the cortex induces an increase in galaninergic markers in the nbm, a result that might suggest that the galaninergic overexpression described in the basal forebrain of patients with Alzheimer's disease can result from a degeneration of the cholinergic basalocortical system from the cortex. Because galanin was found to reduce the activity of the basalocortical cholinergic system only when this one is activated, galanin might exert its role rather during activation deficits than under resting conditions such as the resting cortical hypometabolism, which is characteristic of Alzheimer's disease. PMID- 15545922 TI - Cerebral hemodynamic reserve and early neurologic deterioration in acute ischemic stroke. AB - Early neurological deterioration (END) is associated with increased mortality and morbidity. Although several predictive factors have been reported, there are little data about the hemodynamic factors. Our aim was to determine the capacity of cerebral hemodynamic reserve (CHR) to predict END. We studied 100 hospitalized patients with a first ever ischemic stroke of the middle cerebral artery (MCA) within the first 24 hours of symptoms onset. END was defined as a drop of at least one point in the Canadian Stroke Scale between admission and 72 hours. The mean flow velocity (mV) in the MCA and the CHR were measured by means of transcranial Doppler within the first 24 hours of admission. The CHR was expressed as the percentage increase in the MCA mV divided by the absolute increase in the end-tidal CO2 pressure in mm Hg after carbogen inhalation. END was observed in 23 patients. Reduced values of the mV in the symptomatic MCA (P = 0.043) and of the CHR in the symptomatic hemisphere (P < 0.001) were significantly associated with END. A CHR of less than 2%/1 mm Hg was independently associated with END (OR 8.45, 95% CI 1.82-39.2) after adjusting for potential confounders. CHR impairment within the first 24 hours of acute ischemic stroke is associated with a higher risk of END. This technique may be useful in selecting patients requiring a more intensive management. PMID- 15545923 TI - Activation of proinflammatory caspases by cathepsin B in focal cerebral ischemia. AB - Cathepsins and caspases are two families of proteases that play pivotal roles in ischemic cell death. This study investigated the existence of a cross-talk between cathepsin B and proinflammatory caspases in stroke-induced cell death, as recently suggested by in vitro data. Cortical ischemic damage was induced in mice by distal and permanent occlusion of the middle cerebral artery. Cytoplasmic activation of cathepsin B was observed from the early stages of infarction, and displayed an activation pattern parallel to the activation pattern of caspase-1 and -11. Immunohistochemistry revealed the colocalization of cathepsin B with each caspase in cells of the infarct core. The apical position of cathepsin B in both caspase-activation cascades was confirmed by pretreatment of the animals with the cathepsin B inhibitor CA-074, which also potently protected cortical structures from ischemic damage, indicating involvement of the proteases in the lesion process. The results show that cathepsin B release is an early event following occlusion of cerebral arteries, which eventually triggers the activation of proinflammatory caspases in the absence of reperfusion. This new pathway may play a critical role in brain infarction by promoting inflammatory responses, and/or by amplifying the apoptotic process. PMID- 15545924 TI - Retroviral delivery of homeobox D3 gene induces cerebral angiogenesis in mice. AB - Angiogenesis is regulated by concerted actions of angiogenic and angiostatic factors. Homeobox D3 gene (HOXD3) is a potent proangiogenic transcription factor that promotes angiogenesis by modulating the expression of matrix-degrading proteinases, integrins, and extracellular matrix components. Application of HOXD3 can promote angiogenesis in the skin, but its role in other vascular beds has not been examined. The authors examined HOXD3 expression in human brain vessels by in situ hybridization. Although little or no HOXD3 mRNA was detected in normal brain vessels, increased levels of HOXD3 and its target gene, alpha V beta 3, were found in angiogenic vessels in human brain arteriovenous malformations. The authors further investigated whether HOXD3 plays a role in cerebral angiogenesis in a murine model. Expression of HOXD3 in mouse brain was achieved through retroviral vector-mediated HOXD3 gene transfer. HOXD3 expression lead to a significant induction of cerebral angiogenesis as shown by quantitative microvessel counting (HOXD3: 241 +/- 19 vessels/mm2 vs. saline: 150 +/- 14 vessels/mm2, P < 0.05). The data also showed that focal cerebral blood flow was increased in the angiogenic region with less vascular leakage. Moreover, expression of HOXD3 led to an increase in the expression of a direct downstream target gene alpha V beta 3 integrin. The data suggest that HOXD3 may play an important role in regulating cerebral angiogenesis, and that gene transfer of HOXD3 may provide a novel and potent means to stimulate angiogenesis. PMID- 15545925 TI - Inhibition of toll-like receptor and cytokine signaling--a unifying theme in ischemic tolerance. AB - Cerebral ischemia triggers acute inflammation, which exacerbates primary brain damage. Activation of the innate immune system is an important component of this inflammatory response. Inflammation occurs through the action of proinflammatory cytokines, such as TNF, IL-1 beta and IL-6, that alter blood flow and increase vascular permeability, thus leading to secondary ischemia and accumulation of immune cells in the brain. Production of these cytokines is initiated by signaling through Toll-like receptors (TLRs) that recognize host-derived molecules released from injured tissues and cells. Recently, great strides have been made in understanding the regulation of the innate immune system, particularly the signaling mechanisms of TLRs. Negative feedback inhibitors of TLRs and inflammatory cytokines have now been identified and characterized. It is also evident that lipid rafts exist in membranes and play a role in receptor mediated inflammatory signaling events. In the present review, using this newly available large body of knowledge, we take a fresh look at studies of ischemic tolerance. Based on this analysis, we recognize a striking similarity between ischemic tolerance and endotoxin tolerance, an immune suppressive state characterized by hyporesponsiveness to lipopolysaccharide (LPS). In view of this analogy, and considering recent discoveries related to molecular mechanisms of endotoxin tolerance, we postulate that inhibition of TLR and proinflammatory cytokine signaling contributes critically to ischemic tolerance in the brain and other organs. Ischemic tolerance is a protective mechanism induced by a variety of preconditioning stimuli. Tolerance can be established with two temporal profiles: (i) a rapid form in which the trigger induces tolerance to ischemia within minutes and (ii) a delayed form in which development of protection takes several hours or days and requires de-novo protein synthesis. The rapid form of tolerance is achieved by direct interference with membrane fluidity, causing disruption of lipid rafts leading to inhibition of TLR/cytokine signaling pathways. In the delayed form of tolerance, the preconditioning stimulus first triggers the TLR/cytokine inflammatory pathways, leading not only to inflammation but also to simultaneous upregulation of feedback inhibitors of inflammation. These inhibitors, which include signaling inhibitors, decoy receptors, and anti inflammatory cytokines, reduce the inflammatory response to a subsequent episode of ischemia. This novel interpretation of the molecular mechanism of ischemic tolerance highlights new avenues for future investigation into the prevention and treatment of stroke and related diseases. PMID- 15545926 TI - [Forced voice and its complications]. PMID- 15545927 TI - [Meningo-encephalic complications of chronic cholesteatomatous otitis]. PMID- 15545928 TI - [Anatomohistologic study of von Oort's vestibulocochlear anastomosis]. AB - OBJECTIVES: The vestibulocochlear anastomosis was first described in 1918 by von Oort. It is situated deeply at the bottom of the internal acoustic meatus, and spreads from the saccular nerve before its terminal ramifications, to the cochlear nerve before its penetration into the cochlea. Nerve fibers of the cochlear efferent system are thought to pass through it. The aim of our study was to investigate the anatomy of the vestibulocochlear anastomosis and characterize its histological features. METHOD: [corrected] Ten human temporal bones were dissected. Serial sections were obtained for histological evaluation. RESULTS: The vestibulocochlear anastomosis was found in seven of the specimens, perfectly visualized in six. Average diameter was 0.5 mm with lengths varying from 0.5 to 1 mm. Serial histological sections demonstrated the nervous nature of the anastomosis and its relations with the saccular and cochlear nerves. The epinevrium of the saccular nerve was continuous with the supposed anastomosis in five of the specimens, demonstrating the distinct nature of the anastomosis from the saccular and cochlear nerves. We did not find any evidence linking these fibers to the cochlear efferent system. DISCUSSION: The vestibulocochlear anastomosis was found in seven of our ten dissections. The anastomosis is probably an anatomic reality composed of nerve fibers. The efferent function of these fibers remains to be demonstrated. PMID- 15545929 TI - [Ethmoid adenocarcinoma: trans-facial approach for anterior skull base resection. a series of 80 cases]. AB - OBJECTIVE: To present results of a retrospective analysis of eighty cases of ethmoid adenocarcinoma. Carcinologic and surgical results of anterior skull base resection via the transfacial approach are presented. METHODS: Tumors were classified as 5% T1, 23% T2, 31% T3, 21% T4a and 20% T4b. Thirty-four patients were treated via a paralateronasal approach without skull base resection. Anterior skull bas resections were performed via the transfacial approach for 26 patients and by combined neurosurgical approach for 21. RESULTS: Mean follow-up was 4.8 years. Survival rate was 63.4% at 5 years and 57.9% at 8 years. Forty-two patients were alive and disease-free at last follow-up. Three patients were alive with recurrence. The rate of local recurrence was 38.8%. Complications occurred in 20% of the patients who had a transfacial approach. Complications appeared to be less frequent than with the combined approach. CONCLUSION: Prognosis is related to local control and could be improved by using skull base resection more systematically. In our experience this can be managed by a transfacial approach with similar carcinological results and less complications than the combined approach. PMID- 15545930 TI - [Endoscopic closure of septal perforations by mucosal rotation flaps]. AB - OBJECTIVES: We present a technique of endoscopic endonasal closure of average sized (0.5 - 2 cm) to large (>2 cm) septal perforations. MATERIAL AND METHODS: The surgical technique involves a mucosal rotation flap on a unilateral posterior pedicle without interposition material. We reviewed our experience with eleven patients with average sized and large septal perforations treated over an 8-year period. RESULTS: We based our evaluation on the model proposed by Younger and Blokmanis. Our results were comparable with earlier publications. One-phase closure was achieved in 75% of patients (55% to 90% in the literature reporting all techniques, including external septorhinoplasty and midfacial degloving). CONCLUSION: Our preliminary series provides a basis for a new approach to this condition. The technique is promising for large perforations. This technique could also be considered for other nasal fossae in the event of failed closure. It also offers a way to obtain excellent closure of the anterior portion of the perforation, relieving the patient of the most annoying symptoms. The technique does not contraindicate other procedures which may be performed during the same operation or later. PMID- 15545931 TI - [Mitomycin C: prevention and treatment of anterior glottic synechia]. AB - OBJECTIVES: Topical applications of mitomycin C to the anterior glottis may prevent anterior glottic synechia (prevention group) or restenosis (treatment group). METHODS: In the prevention group, six patients with glottic carcinoma involving the anterior commissure were treated by transoral laser surgery. Repeated procedures were performed in one patient. For the six patients in the treatment group, the anterior glottic synechia was secondary to frontolateral laryngectomy (three patients), transoral laser therapy for laryngeal papillomatosis (two patients) or bilateral glottic carcinoma (one patient). Mitomycin C (0.4 mg/ml) was used as a topical application on the anterior commissure for a duration of 4 minutes. Outcome was assessed clinically at three months using a visual scale: no synechia (success), micro-synechia (partial failure), and synechia (failure). RESULTS: In the prevention group, there were six successes and one partial failure. In the treatment group, there were two successes, three partial failures, and one failure. No side effects were noted. CONCLUSION: Topical application of mitomycin C was effective to prevent anterior glottic synechia after transoral laser surgery for glottic carcinoma involving the anterior commissure. It is an alternative to endolaryngeal keel in patients with sequellar synechia. These preliminary results should be further evaluated in a larger series. PMID- 15545932 TI - [Interest of PET/CT scan fusion to assess mandible involvement in oral cavity and oro pharyngeal carcinomas]. AB - The aim of the study is to assess mandible involvement in oral cavity and oropharyngeal carcinomas. OBJECTIVE: To evaluate interest of fusion of Positron Emission Tomography (PET) with Computed Tomography scan (CT scan). METHOD: Eight patients were included in this prospective study. Each patient underwent PET and CT scan of the head and neck before surgery including tumorectomy and mandibulectomy. We compared results of PET- CT fusion with histologic examinations. RESULTS: Oral cavity (6), oropharyngeal (2) carcinoma: Mandibular invasion was suspected by PET-CT in 3 cases, but was confirmed in histological examination in only 2 cases. In 5 cases, PET-CT did not find mandibular invasion; this was confirmed in histological examination in all cases. Sensibility of PET CT fusion was 100%, specificity was 83%. Positive predictive value was 66% and negative predictive value was 100%. DISCUSSION: PET-CT fusion provided maximal sensitivity. Specificity was better than for MRI but less than CT-scan. There were no false negatives and the false positive rate was 33%. CONCLUSION: PET-CT fusion is interesting to predict mandible involvement. Further studies are necessary to confirm these preliminary results. PMID- 15545933 TI - [Metachronous sinonasal squamous-cell carcinoma after pharyngolaryngectomy]. AB - OBJECTIVE: Three laryngectomized patients developed metachronous sinonasal squamous-cell carcinoma. We reviewed their files to search for clinical features useful for early diagnosis of this localization. CASE REPORTS: Two of the patients, a textile worker and a food processing worker, had occupation exposure risk factors for sinonasal squamous-cell carcinoma. The anatomic modifications created by laryngectomy contributed to late diagnosis of the metachronous tumor at an advanced stage. Surgery was performed in all three patients. Local recurrence was observed at one year in two patients. DISCUSSION: Sinonasal fibroscopy should be part of the surveillance scheme in laryngectomized patients to enable early diagnosis and treatment of metachronous tumors. PMID- 15545934 TI - [Extracranial internal carotid artery aneurysm]. AB - INTRODUCTION: Extracranial aneurysm of the internal carotid artery is an exceptional finding. CASE REPORT: An 89-year-old woman consulted for a peritonsillar mass. Physical examination revealed a parapharyngeal pulsatile mass in the oropharynx. Computed tomography (CT) provided the diagnosis of extracranial internal carotid artery aneurysm. Endovascular or surgical treatment were declined. Anticoagulation medication was given. DISCUSSION: We reviewed the CT and magnetic resonance imaging findings and the clinical manifestations of extracranial internal artery aneurysm. Endovascular treatment is an alternative to open surgery. PMID- 15545936 TI - [Neurologic emergencies]. AB - Neurological symptoms are a very frequent cause of consultation in emergency units and require consultation with neurologists and neuroradiologists. The most frequent diagnoses are stroke syndrome, seizure, headache, confusion, meningitis and meningo-encephalitis, and facial palsy. The morbidity and mortality of neurological emergencies are strongly related to prompt medical management of the patients which often requires neuroimaging studies. The most common neurological emergencies will be reviewed. PMID- 15545937 TI - [Evaluation of neurological deficits]. AB - The first step in the evaluation of patients with neurological deficit is to obtain expert clinical evaluation to achieve accurate and complete characterization. The type of neurological deficit, its mode of onset, the presence or absence of associated signs and symptoms, and a differential diagnosis must first be described. This will allow identification of the probable site of CNS involvement and appropriate imaging evaluation to be performed. A multi-disciplinary approach with constant correlation between clinical, imaging and pathological findings will enable clinicians to provide patients accurate diagnoses, appropriate prognostic evaluation and optimal management. PMID- 15545938 TI - [Headaches in the emergency context]. AB - Headaches constitute one of the most frequent reason of consultation. Their causes are extremely varied. The first step consists in the analysis of the characteristics of the pain and the associated signs in order to distinguish primary and secondary headaches. Primary headaches, including migraines and tension-type headaches are the most frequent types and do not require imaging evaluation. Secondary headaches are related to an organic cause and require specific investigations. In case of suspected symptomatic or secondary headaches, brain imaging plays an important role in the etiologic work-up. The main purpose of imaging in an emergency setting is to diagnose a life-threatening disease. PMID- 15545939 TI - [Pediatric neuroimaging emergencies]. AB - The notion of emergency with regards to pediatric neuroimaging requires a strong knowledge of clinical indications. In children under 2 years of age, head trauma requires a CT scan in case of repeated or prolonged or rapidly increasing vomiting, focal signs, loss of consciousness, unusual behavior, seizures, clinical signs of skull fracture or polytrauma. The "shaken baby syndrome" is usually suspected in case of loss of consciousness or seizures before 8 months of age. The hematomas that are observed are subdural in location, diffuse and deeply located. Imaging is only mandatory for headache suggesting underlying space occupying lesion: permanent or increasing pain, nocturnal headache, headache during postural changes or efforts, associated to seizures or abnormal neurological examination. No imaging is indicated in case of first epileptic seizure associated to normal neurological examination and without any particular context. The presence of trauma, intracranial hypertension, persisting disturbances of consciousness or associated focal sign necessitates urgent neuroimaging. No imaging is indicated in case of typical febrile seizures, i.e. generalized, brief and occurring between 1 and 5 years of age. Spinal cord symptoms require immediate MRI evaluation. The most frequent tumor is neuroblastoma. In the absence of spinal tumor, brain abnormalities must be excluded (inflammatory disease). In neonates, CT scan or MRI must be readily performed in case of seizures or loss of consciousness to exclude ischemic, traumatic or infectious lesions. PMID- 15545940 TI - [Management of patients after a first seizure]. AB - Neuroimaging evaluation in patients after a first seizure could be easily determined on the basis of seizure history, neurological examination, blood sample analysis and electroencephalography. The main objectives of the initial work-up are to differentiate a true seizure event from seizure-like symptoms, to exclude a single seizure as a manifestation of non organic cause and finally to consider the seizure as a result of cerebral lesion or inaugurate epilepsy. When a new onset seizure is diagnosed, urgent neuroimaging is recommended only in patients with focal neurological deficit, persistent or worsening alteration in the level of consciousness and when clinical and biological data lead to a suspected vascular or infectious etiology. Brain CT scan is usually more available in emergency to identify the cause of seizure. It may have an important role for the therapeutic strategy and may defer MRI investigation. Nevertheless, brain MRI must be performed in emergency when CT scan is not conclusive despite a severe clinical condition or in case of cerebral venous thrombosis. Imaging modalities depend on clinical data, patient age and suspected epilepsy type. PMID- 15545941 TI - [Imaging of neuro-ophthalmological emergencies]. AB - MRI often is mandatory in the diagnostic work-up of visual loss, visual field alterations and oculomotor problems. It is performed emergently in patients with painful diplopia associated to mydriasis, to exclude aneurysm, or in patients with painful Horner syndrome to exclude dissection of the internal carotid artery. CT scan in emergency remains useful in case of acute lateral hemianopsia or acute post traumatic visual loss. Progressive neuro-ophthalmological symptoms may require imaging examination in a short delay to define the therapeutic strategy: monocular transient blindness (dissection or carotid stenosis), progressive visual loss (optic nerve compression), bitemporal hemianopsia (optic chiasm lesion), painful visual loss (optic neuritis). A very precise clinical indication is helpful for the choice of imaging protocol and to improve its diagnosis value. PMID- 15545942 TI - [Neuroradiologic emergencies in infectious pathology]. AB - The main goal of urgent imaging evaluation of patients with suspected CNS infection is to differentiate infectious from tumoral or vascular lesions in order to provide appropriate management. MR imaging, including diffusion weighted imaging and spectroscopy, is superior to CT imaging to characterize lesion location and etiology. The CT and MRI features of the more frequent bacterial, viral and parasitic CNS infections will be described. PMID- 15545944 TI - [Emergency imaging of cerebrovascular accidents]. AB - Over the last 25 years, advances in neuroimaging have significantly changed the evaluation and management of acute stroke syndromes. In the seventies, computed tomography (CT) could differentiate between ischemic and hemorrhagic stroke. Magnetic resonance imaging (MRI) is nowadays the imaging modality of choice in the initial assessment of acute stroke. MRI images can better discriminate acute, subacute and chronic infarcts, differentiate venous from arterial infarcts, detect arterial dissection, stenosis or occlusion. Diffusion-weighted images are highly sensitive and specific to acute infarction and the combination with perfusion technique is suitable to define potentially reversible ischemia (area of cerebral "mismatch" which is thought to represent the so-called ischemic penumbra). This penumbra is a potential therapeutic target of valuable interest for the treating physician. PMID- 15545943 TI - [Imaging in acute toxic encephalopathy]. AB - Neuroimaging, particularly MR imaging, plays a major role for the diagnosis of many acute toxic encephalopathies. Toxic disorders are related to drugs (immunosuppressive agents, chemotherapeutic agents, anti-epileptic drugs, heroin...), to metals (lead, manganese, mercury...), and to industrial and environmental chemicals (solvent, carbon monoxide...). MR imaging with diffusion and perfusion imaging provides information regarding brain lesions induced by the toxic agents (vasogenic edema, cytotoxic edema, infarction, hemorrhage, demyelination...). PMID- 15545945 TI - [The value of teleradiology in the management of neuroradiologic emergencies]. AB - First, to summarize the results of teleradiology programs on neurosurgical emergency care in France. Second, to compare French data with the international literature. Third, to discuss the likely developments and future of teleneuroradiology and teleneurosurgery. Data on French use of telemedicine applications in neuroradiology come from a survey of telemedicine applications in France, which has been conducted in year 2003 at the request of the French ministry of the Research. Teleradiology clearly has a positive impact on emergency neurosurgical care by reducing the time to correct diagnosis and initiation of treatment of patients who need to be transferred and avoid unnecessary transfers. However, present teleradiology applications have organizational limitations that are summarized and discussed with reference to the literature. Further developments in information and communications technology have the potential to revolutionise neurosurgical emergency care and contribute to improve the training of neuroradiology and neurosurgery staff. PMID- 15545946 TI - [Medicolegal aspects in neuroradiologic emergencies]. AB - Naturally, neuroimaging emergencies are a component of the global subject of emergencies that have for several years been a frequent subject in the press. The current situation is the consequence of the evolution of ideas from contractual liability to the frequently discussed principle of precaution. Examples offered by the daily practice allow to emphasize the main difficulties: diagnosis of headaches, seizure episode on public roads, head trauma, stroke... The medical liability of the neuroradiologist is not the same for diagnostic or interventional neuroradiology; it can sometimes be shared with other physicians. This liability includes the indication, the quality of the diagnosis, the information of the patient and relatives, the communication of results with other physicians, the maintenance of the equipment... Knowledge of potential pitfalls allows preventive measures including competence of the neuroradiologist, informed patient consent, and written precise protocols are the most important. However, according to the evolution of our societies, it is likely that it will become increasingly difficult to achieve a professional neuroradiology career without being questioned; it is thus advisable to be prepared for such eventuality. PMID- 15545948 TI - [Limitations of drug prescriptions in patients with chronic obstructive pulmonary disease]. AB - National and international guidelines have described in detail the role of bronchodilators and inhaled corticosteroids for the management of patients with chronic obstructive lung disease (COPD). Such guidelines are identical to marketing approval documents in France. The French marketing approval, which limits use of bronchodilators to reversible COPD, is no longer coherent with recent evidence demonstrating the poor reproducibility of reversibility tests and the absence of correlation between reversibility and dyspnea relief. France has not delivered marketing authorization for any inhaled corticosteroid for the treatment of COPD. Clinical trials versus placebo have demonstrated that inhaled corticosteroids do not slow down the decline in ventilatory function but that they do diminish the risk of exacerbation. Decreased risk of exacerbation could be clinically pertinent in patients with severe COPD who often experience exacerbations requiring to specialist management. Inquiries concerning precriptions for COPD show that the majority of patients are treated by inhaled corticosteroids, demonstrating the gap between medical practice and national and international recommendations and marketing approval documents. PMID- 15545947 TI - [Pulmonary hypertension: from genetics to treatments]. AB - Pulmonary hypertertension (PHT) is a rare disease defined by increased resistance of the pulmonary arteries inevitably leading to right heart failure if specific treatment is not given. This disease can occur sporadically (idiopathic or primary PHT), within a familial context (familial PHT, BMPR2 gene mutation), or occur as a complication of other diseases (connective tissue disease, congenital cardiomyopathy, human immunodeficiency virus infection, portal hypertension, use of anorexigenic agents). The incidence of primary PHT is 2 million cases per year, probably an underestimation due to the low specificity of clinical signs, predominantly exercise-induced dyspnea. Recent therapeutic advances (prostacyclin and endothelin receptor antagonists administered in continuous infusion) have improved the prognosis of this orphan disease. Inhaled iloprost and type 5 phosphodiesterase inhibitors should be evaluated for this indication. Lung transplantation is reserved for patients unresponsive to medical treatment. PMID- 15545949 TI - [Efficacy and acceptability of the fixed fluticasone + salmeterol combination in the treatment of acute asthma attacks. Results of a one-year comparative study]. AB - BACKGROUND: The objective of an asthma treatment is to control asthma, particularly to prevent exacerbations. OBJECTIVE: To study the efficacy, acceptability and safety of the fluticasone/salmeterol (FP/S) combination in preventing asthma exacerbations in comparison with the continuation of previous treatment (TA). METHODS: This was a multicentre, randomised, parallel-group study to compare the fixed combination FP/S to TA (treatment with a free combination of an inhaled corticosteroid and a long-acting beta2-agonist) over one year in patients whose asthma was well controlled with their current treatment. RESULTS: Five hundred and twenty patients were randomized and their data analyzed on an intent-to-treat basis. Seventy-four percent of the patients in the FP/S group and 71% in the TA group had no exacerbation. The 3.12% difference in favor of the FP/S group (90% CI: -3.32% to 9.56%) demonstrated that FP/S was at least as effective TA in preventing asthma exacerbations. Treatment acceptability, evaluated by the patient on visual analog scales (inclusion as part of the daily habits, constraint, simplicity) was better with FP/S (p<0.001) than with TA. Clinical safety was good and comparable in the two groups. CONCLUSION: The fixed fluticasone/salmeterol combination provided a protection as good as that obtained with free combinations over a one-year treatment period, with a significant improvement in treatment acceptability and a good clinical safety. PMID- 15545950 TI - [Smoking in the African setting (Abidjan, Ivory coast): patient knowledge, attitude and behavior]. AB - Recent studies on smoking conducted in the ivory coast have been sparse despite increasingly aggressive tobacco company campaigns. The purpose of this prospective study was to examine the influence of medical specialty on patient knowledge, attitude and smoking behavior among people consulting three university hospitals in Abidjan. The inquiry was performed with a questionnaire concerning a comparative series of 180 subjects (90 patients in each sample). Overall, the series included 16.10% active smokers, 15.60% former smokers, and 68.30% non smokers, with no difference between the groups. The well-known feature of smoking with friends was main reason cited for beginning smoking. Three quarters of patients (75.9%) had attempted to stop smoking. Information about smoking was obtained from the media. The hospital was involved little in acquiring knowledge about smoking. Type of specialist consultation (hospital units providing care for smoking-related disease or not) had little influence. Hospital units should become more involved in the anti-smoking campaign. Consultations on cessation of smoking should be instituted in a country with highly aggressive smoking advertisements. PMID- 15545951 TI - [An exceptional but serious complication of benign intercostal schwannoma: hemothorax]. AB - Spontaneous serious hemothorax has not been described previously as an inaugural signor secondary complication of benign intercostal schwannoma. We report a case in a 45-year-old woman who was hospitalized in an emergency setting after development of massive left hemothorax. After evacuation of the effusion, imaging demonstrated a voluminous apparently neurogenic tumor in an intercostal position. Thoracotomy was required for hemostasis due to persistent bleeding. Tumor resection was performed. At histology, the surgical specimen was found to be a benign schwannoma presenting hemorrhagic remodeling. Resection provided complete cure with no sequelae. This secondary complication favors resection of benign intercostal schwannoma. PMID- 15545952 TI - [An unusual diagnosis of post-traumatic hemothorax]. AB - We report the case of malignant pleural mesothelioma revealed by a post-traumatic hemothorax. PMID- 15545953 TI - [Spontaneous pneumothorax revealing malignant pleural mesothelioma. Three case reports]. AB - Spontaneous pneumothorax is an uncommon inaugural presentation of malignant pleural mesothelioma. We report three cases in men aged 65, 30 and 76 years. The diagnosis was suggested at medical imaging and was confirmed at histological analysis of biopsies obtained by thoracoscopy in two patients and thoracotomy in one. The first patient (age 65 years) died two months after the initial diagnosis. The second patient (age 30 years) was alive 40 months after 15 chemotherapy cycles using a platinim-gemcitabine combination. Complete tumor response was achieved in the third patient (age 76 years) after 9 chemotherapy cycles with the same combination. Since mid-term prognosis is fatal for this type of tumor, we propose thoracoscopy in all patients over 30 years who develop spontaneous pneumothorax with no morphological features increasing the risk of pneumothorax, and particularly in patients with asbestos exposure. PMID- 15545955 TI - [Hyperplasia of the thymus in Graves' disease. A case report]. AB - Graves' disease is rarely associated with hypertrophy of the thymus which takes on a pseudotumor aspect. We observed a case in a 40-year-old woman who presented clinical and biological hyperthyroidism with anti-TSH receptor antibodies, favoring the diagnosis of Graves' disease which was confirmed by scintigraphy. The chest x-ray performed because of chest pain revealed enlargement of the mediastinum. The thoracic CT-scan without contrast injected showed a pseudotumor aspect of the thymus. Medical treatment with anti-thyroid drugs and beta blockers led to regression of the thymus mass. Knowledge of this type of association can avoid unnecessary thymus surgery. The clinical course is favorable irrespective of the type of anti-thyroid drug given. PMID- 15545954 TI - [Solitary fibrous mediastinal tumor. A case report]. AB - Solitary fibrous tumor is a rare tumor, generally located in the pleura. Recently, new sites have been described in the literature, particularly involving the peritoneum and mediastinum, as wekk as te-the bronchopulmonary and orbital regions. The diagnosis is established at pathology, but in some cases may be difficult to differentiate from other tumors. We report the case of a 60-year-old women who had undergone surgery 25 years ago for right pulmonary hydatid cyst and who presented inspiratory dyspnea and dry cough for four months. Physical examination revealed a right cervical tumefaction in the supraclavian region. The AP chest x-ray disclosed an opacity in the right laterotracheal upper cervicomediastinal zone laminating the trachea which was displaced to the left. Thoracic computerized tomography showed a tissular process in the superior and middle mediastinum. Surgical tumor resection was performed and pathology confirmed the diagnosis of solitary fibrous tumor. The postoperative period was uneventful. Frequency of the mediastinal localisation is the same in men and women. The tumor generally develops between the 5th and 7th decades, and is most often asymptomatic. Fortuitous discovery is the rule. The clinical course is unforeseeable. PMID- 15545956 TI - [Lung carcinosarcoma. A case report]. AB - Carcinosarcoma of the lung is a rare highly malignant tumor of unknown histogenesis. There is an epithelial carcinomatous component and a malignant mesenchmatous component with heterologous tissue. Survival at two years after surgical resection is not greater than 10%. We report a case observed in a 71 year-old man who developed chest pain. Outcome was fatal at three months. Pathology examination of a transparietal biopsy provided the diagnosis which was confirmed by immunohistochemistry on the surgical resection specimen. PMID- 15545957 TI - Rapid determination of Epstein-Barr virus latent or lytic infection in single human cells using in situ hybridization. AB - Epstein-Barr (EBV) virus is associated with malignancies such as lymphoma and carcinoma. Infection of cells with EBV may result in either lytic infection with production of viral particles, characterized by the presence of linear DNA forms, or latent infection, characterized by either episomal or integrated DNA forms. To examine whether the different lytic and latent EBV DNA forms can reliably be distinguished in single human cells, in situ hybridization was performed in EBV positive cell lines. Immunocytochemistry and Southern blot analysis were performed supplementary to in situ hybridization. In latent infection, three in situ hybridization patterns were observed: large-disperse (episomal), small punctate (integrated) and combined (both), signal types 1, 2 and 3 respectively. These were associated with expression of latent membrane protein 1, but not with Z fragment of Epstein-Barr replication activator or viral capsid antigen. In lytic infection, three additional in situ hybridization patterns were observed: nuclear membrane associated, bubble (filling up the nucleus) and spillover (covering the lysed cells) signals types 4, 5 and 6 respectively. Signal types 4 and 5 were associated with expression of latent membrane protein 1 and Z fragment of Epstein-Barr replication activator but not viral capsid antigen, whereas type 6 was associated with expression of viral capsid antigen only. Southern blot analysis confirmed these results; however, low copy numbers of integrated virus were often missed by Southern blot, confirming that in situ hybridization is more sensitive in determining the presence of all types of EBV DNA. In situ hybridization may prove useful in rapidly screening large series of tissue microarrays and other clinical specimens for the presence of lytic or latent EBV. PMID- 15545958 TI - Choriocarcinoma involving the pancreas as first manifestation of a metastatic regressing mixed testicular germ cell tumor. AB - We describe an unusual case of metastatic choriocarcinoma of the pancreas arising from a regressing testicular mixed germ cell tumor that clinically mimicked a primary pancreatic tumor. A 54-year-old male presented with a 2-month history of progressive upper abdominal pain, weight loss, and jaundice. He also had a history of recurrent epididymitis associated with the presence of a right testicular mass shown to be cystic by ultrasound and stable for at least 10 years. A computed tomography scan showed an isolated 6 cm mass in the head of the pancreas. A pancreaticoduodenectomy was performed. Upon histological examination, the pancreatic tumor showed extensive hemorrhage and necrosis. In the viable area, the tumor was composed of an intimate mixture of mononuclear cytotrophoblast cells and multinucleated syncytiotrophoblasts with vascular invasion. These characteristic features led to the correct diagnosis on frozen section. The cytology of the tumor was nonspecific and suggested undifferentiated carcinoma of the pancreas. The trophoblastic origin of the tumor cells was confirmed by immunohistochemistry staining. The testicular mass showed a regressed mixed germ cell tumor of predominantly seminoma with focal teratoma but without a choriocarcinoma component. In conclusion, we present a rare and unusual case of a regressing testicular mixed germ cell tumor that presented as a primary pancreatic tumor. Cytological features of the pancreatic mass were not specific and raised the possibility of a primary undifferentiated carcinoma of the pancreas. Characteristic histological features of choriocarcinoma led to the correct diagnosis on frozen section. Subsequent resection of the testicular mass confirmed the presence of a cystic and scarring (regressing) mixed germ cell tumor but without evidence of choriocarcinoma. PMID- 15545964 TI - Tissue microarrays, tread carefully. PMID- 15545965 TI - Additive interaction of gefitinib ('Iressa', ZD1839) and ionising radiation in human tumour cells in vitro. AB - Cultures of human carcinoma A-431, A-549 and HeLa cells were challenged with gamma-rays without or with concomitant exposure to gefitinib, a potent inhibitor of the tyrosine kinase activity of epidermal growth factor receptor (EGFR). The outcome of treatment was determined from cell and colony count, cell cycle progression and DNA double-strand break formation and rejoining. Apoptosis was measured in parallel from hypodiploid DNA and using an annexin V assay. Gefitinib developed a cytostatic effect in all cell lines, with drug sensitivity correlating the level of EGFR expression. A weak cytotoxicity of gefitinib was observed in HeLa cells only, although the drug was unable to induce significant cell cycle redistribution in this cell line. In contrast, substantial G1 block and S-phase depletion was observed in A-431 and A-549 cells exposed to gefitinib. The drug brought about additive to subadditive interaction with radiation with regard to growth inhibition, clonogenic death and induction of apoptosis. Consistently, gefitinib did not hinder the rejoining of radiation-induced DNA double-strand breaks in any cell line. The results demonstrate that gefitinib may elicit cytotoxicity at high concentration, but does not act as a radiosensitiser in vitro in concomitant association with radiation. PMID- 15545966 TI - Oral contraceptive use and ovarian cancer risk among carriers of BRCA1 or BRCA2 mutations. AB - Women with mutations of the genes BRCA1 or BRCA2 are at increased risk of ovarian cancer. Oral contraceptives protect against ovarian cancer in general, but it is not known whether they protect against the disease in carriers of these mutations. We obtained self-reported lifetime histories of oral contraceptive use from 451 women who carried mutations of BRCA1 or BRCA2. We used conditional logistic regression to estimate the odds ratios associated with oral contraceptive use, comparing the histories of 147 women with ovarian cancer (cases) to those of 304 women without ovarian cancer (controls) who were matched to cases on year of birth, country of residence and gene (BRCA1 vs BRCA2). Reference ages for controls had to exceed the ages at diagnosis of their matched cases. After adjusting for parity, the odds-ratio for ovarian cancer associated with use of oral contraceptives for at least 1 year was 0.85 (95 percent confidence interval, 0.53-1.36). The risk decreased by 5% (1-9%) with each year of use (P for trend=0.01). Use for 6 or more years was associated with an odds ratio of 0.62 (0.35-1.09). These data support the hypothesis that long-term oral contraceptive use reduces the risk of ovarian cancer among women who carry mutations of BRCA1 or BRCA2. PMID- 15545967 TI - TP53 status determines clinical significance of ERBB2 expression in ovarian cancer. AB - ERBB2 expression has been found in 19 to 44% of ovarian carcinomas; however, its predictive value has not been demonstrated, and trastuzumab has not found clinical application in ovarian cancer patients. We evaluated clinical significance of ERBB2 expression in relation to TP53 accumulation in ovarian carcinoma patients treated with platinum-based regimens. Immunohistochemical analysis with CB11 and a novel NCL-CBE356 antibody (against the internal and external domains of ERBB2, respectively) was performed on 233 tumours (FIGO stage IIB-IV); the US Food and Drug Administration-approved grading system with 0 to 3+ scale was used for evaluation, and the results were analysed by the Cox and logistic regression models. In all, 42% of the tumours expressed (category 1+, 2+ or 3+) either CB11 or CBE356 or both (CB11/CBE356 parameter). Associations between ERBB2 expression and clinical factors were observed only if tumours with staining category 1+ were grouped together with tumours showing staining categories 2+ and 3+. CB11/CBE356 parameter had a better predictive value than CB11 alone. CB11/CBE356 expression was negatively associated with platinum sensitivity (PS) in the TP53(-) group (P=0.022) and with disease-free survival (DFS) in the TP53(+) group (P=0.009). Our results may suggest that trastuzumab should be given postoperatively to patients with TP53(-)/ERBB2(+) ovarian carcinomas to enhance PS, and after completion of chemotherapy to patients with complete remission and TP53(+)/ERBB2(+) carcinomas to extend DFS time (in total to 30.4% of all patients analysed). Thus, novel criteria for ovarian cancer patient inclusion for clinical trials with trastuzumab should be considered and tested. PMID- 15545968 TI - Genomic imbalances in 70 snap-frozen cervical squamous intraepithelial lesions: associations with lesion grade, state of the HPV16 E2 gene and clinical outcome. AB - Host genomic abnormalities may determine the natural history of cervical squamous intraepithelial lesions (SILs). We undertook comparative genomic hybridisation analysis of epithelium carefully microdissected from 70 cervical SILs, the largest series to date. In contrast to previous studies, we used frozen sections for optimal DNA quality and examined whether patterns of DNA copy number imbalance (CNI) are characteristic of SIL grade, human papillomavirus (HPV) status and postoperative recurrence. We identified more CNIs in cervical SIL than previously described, with more CNIs per case in high-grade squamous intraepithelial lesion (HG-SIL) than in low-grade squamous intraepithelial lesion (LG-SIL) (P=0.04). While some CNIs were seen at similar frequencies in HG-SIL and LG-SIL, others, including gain on 1q, 3q and 16q, were found frequently in HG-SIL but not in LG-SIL. There were significantly more CNIs per case in HG-SILs showing loss of the HPV16 E2 gene (a repressor of viral oncogene transcription) (P=0.026) and in HG-SILs that subsequently recurred (P=0.04). Our data are consistent with sequential acquisition of CNIs in cervical SIL progression. Higher frequency of CNI in association with E2 gene loss supports in vitro evidence that high-risk HPV integration is associated with genomic instability. Further investigation of the clinical value of specific host genomic abnormalities in cervical SIL is warranted. PMID- 15545969 TI - Recent trends in cervical cancer mortality in Britain and Ireland: the case for population-based cervical cancer screening. AB - This study used published mortality data and regression techniques to look at time trends in cervical cancer mortality between 1970 and 2000 in the UK and the Republic of Ireland. Mortality from cancer of the cervix has been declining in the UK for at least the past 30 years. The rate of decrease has been greatest in England, Wales and Scotland and has accelerated in these countries since the reorganisation of screening services in the late 1980s. Mortality in Northern Ireland is also decreasing, but at a lesser rate and without significant change over the same period. In contrast, cervical cancer mortality in the Irish Republic, which, unlike the UK, does not have comprehensive population-based screening, has been increasing by an average of 1.5% per year since 1978. The mortality rate, which was half of that in the UK in the late 1970s, now exceeds that in any of the region of the UK. The absence of population-based screening for cervical cancer in the Republic of Ireland is the most plausible explanation for these differences in trend. PMID- 15545970 TI - Endostatin expression in pancreatic tissue is modulated by elastase. AB - Pancreatic tumours are scirrhous, avascular tumours, suggesting that they may produce angiogenesis inhibitors that suppress the growth of the vasculature to the tumour and metastases. We have sought evidence for the angiogenesis inhibitor, endostatin, in normal and cancerous pancreatic tissue. Using Western blotting, we found mature 20 kDa endostatin in cancer tissue but not in normal tissue. Several endostatin-related peptides of higher mol wt were present in both tissues. Extracts from normal tissue were able to degrade exogenous endostatin, whereas extracts from cancer were without effect. Although the exocrine pancreas secretes inactive proenzymes of trypsin, chymotrypsin and elastase, their possible role in this degradation was examined. The trypsin/chymotrypsin inhibitor, Glycine max, did not prevent the degradation of endostatin by normal pancreatic extracts but elastatinal, a specific inhibitor of elastase, reduced the rate of degradation. Extracts of pancreatic tumours did not express any detectable elastase activity, but an elastase (Km 1.1 mM) was expressed by extracts of normal pancreas. We conclude that endostatin is present and stable in pancreatic cancer tissues, which may explain their avascular nature, but that normal pancreatic tissue expresses enzymes, including elastase, which rapidly degrade endostatin. The stability of endostatin may have implications for its therapeutic use. PMID- 15545972 TI - Laminin-5 offsets the efficacy of gefitinib ('Iressa') in hepatocellular carcinoma cells. AB - Prognosis and survival of patients with hepatocellular carcinoma (HCC) is still very poor, and no therapies are currently available to inhibit tumour growth and metastases. Recently, we reported that the expression of an extracellular matrix component (ECM), namely Laminin-5 (Ln-5), is directly related to poor prognosis in HCC patients. The aim of our study is to investigate the preclinical effect of gefitinib in an in vitro HCC model. We found that the IC(50) of gefitinib in HCC cells ranged from 0.7 to 10.0 muM, whereas Ln-5 inhibited the activity of gefitinib in a dose-dependent manner. Complete inhibition of phosphorylated (p) EGFR (epidermal growth factor receptor) was obtained within 6 h exposure to gefitinib and complete restoration of the receptor status was obtained after 24 h. A downstream effect yields a decrease in p-Akt and p-Erk 1/2. The addition of exogenous Ln-5 has no effect on p-EGFR, whereas it restores p-Erk 1/2 and p-Akt. Consistently, Ln-5 induces recovery of HCC cells from Gefitinib-induced apoptosis. In conclusion, gefitinib inhibits HCC cell growth and we report for the first time that Ln-5, but not other ECM molecules, reduces the ability of gefitinib to inhibit cell growth via Akt. As patients with HCC have different Ln 5 expression levels, these results may help to better understand which patients might benefit from gefitinib treatment. PMID- 15545971 TI - Ki-67 expression and patients survival in lung cancer: systematic review of the literature with meta-analysis. AB - Among new biological markers that could become useful prognostic factors for lung carcinoma, Ki-67 is a nuclear protein involved in cell proliferation regulation. Some studies have suggested an association between Ki-67 and poor survival in lung cancer patients. In order to clarify this point, we have performed a systematic review of the literature, using the methodology already described by our Group, the European Lung Cancer Working Party. In total, 37 studies, including 3983 patients, were found to be eligible. In total, 49% of the patients were considered as having a tumour positive for the expression of Ki-67 according to the authors cutoff. In all, 29 of the studies dealt with non-small-cell lung carcinoma (NSCLC), one with small-cell carcinoma (SCLC), two with carcinoid tumours and five with any histology. In terms of survival results, Ki-67 was a bad prognosis factor for survival in 15 studies while it was not in 22. As there was no statistical difference in quality scores between the significant and nonsignificant studies evaluable for the meta-analysis, we were allowed to aggregate the survival results. The combined hazard ratio for NSCLC, calculated using a random-effects model was 1.56 (95% CI: 1.30-1.87), showing a worse survival when Ki-67 expression is increased. In conclusion, our meta-analysis shows that the expression of Ki-67 is a factor of poor prognosis for survival in NSCLC. PMID- 15545973 TI - Quantifying trade-offs: quality of life and quality-adjusted survival in a randomised trial of chemotherapy in postmenopausal patients with lymph node negative breast cancer. AB - We evaluated quality of life (QL) and quality-adjusted survival in International Breast Cancer Study Group Trial IX, a randomised trial including 1669 eligible patients receiving tamoxifen for 5 years or three prior cycles of cyclophosphamide, methotrexate and 5-fluorouracil (CMF) followed by 57 months tamoxifen. During the time with CMF toxicity (Tox), without symptoms and toxicity (TWiST), and following relapse (Rel), patients scored their QL indicators and a utility indicator for subjective health estimation between 'perfect' and 'worst' health. Scores were averaged within Tox, TWiST and Rel and transformed to utilities. Mean durations for the three transition times were weighted with utilities to obtain mean quality-adjusted TWiST (Q-TWiST). Patients receiving CMF reported significantly worse scores for most QL domains at month 3, but less hot flushes. After completing chemotherapy, there were no differences by treatment groups. Benefits evaluated by Q-TWiST favoured the additional chemotherapy. CMF provided 3 more months of Q-TWiST for patients with ER-negative tumours, but CMF provided no benefit in Q-TWiST for patients with ER-positive tumours. Q-TWiST analysis based on patient ratings is feasible in large-scale cross-cultural clinical trials. PMID- 15545974 TI - Interleukin-8/CXCL8 is a growth factor for human lung cancer cells. AB - Interleukin-8/CXCL8 (IL-8) is a chemokine and angiogenic factor. Recently, IL-8 was identified as an autocrine growth factor in several human cancers. Here, we investigated the expression and function of IL-8 in lung cancer cells. The expressions of IL-8 and its receptors, CXCR1 and CXCR2, were examined in a panel of non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) cell lines. Using reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay, we found that all NSCLC cell lines tested produced modest or high levels of IL-8 (up to 51 ng ml(-1) 10(6) cells(-1)). Expression of CXCR1 and CXCR2 was found by RT-PCR and flow cytometry in two out of three cell lines. In contrast, SCLC cell lines produced very low or undetectable levels of IL-8, but expressed CXCR1 and CXCR2. We next investigated whether IL-8 could act as an autocrine growth factor in two NSCLC cell lines (H460 and MOR/P) expressing both IL-8 and its receptors. We found that cell proliferation was attenuated by anti-IL-8 neutralising antibody to 71 and 76% in H460 and MOR/P, respectively (P<0.05). Exogenous IL-8 significantly stimulated cell proliferation in four SCLC cell lines tested in a dose-dependent fashion. Cell proliferation was increased by between 18% (P<0.05) and 37% (P<0.05). Stimulation of cell proliferation by IL 8 was also demonstrated by analysis of proliferating cell nuclear antigen expression and cell cycle in H69 cells. Furthermore, we investigated which receptor(s) mediated the mitogenic function of IL-8 in lung cancer cells. We found that cell proliferation was significantly reduced by anti-CXCR1 antibody but not by anti-CXCR2 antibody. In conclusion, IL-8 can act as an autocrine and/or paracrine growth factor for lung cancer cells, and the mitogenic function of IL-8 in lung cancer is mediated mainly by CXCR1 receptor. PMID- 15545976 TI - Predictors of drop-out in overweight and obese outpatients. AB - OBJECTIVE: To investigate the impact on drop-out rates of several baseline clinical characteristics of a sample of overweight and obese outpatients. DESIGN: Retrospective clinical trial. SUBJECTS: The charts of 383 patients aged 15-82 y attending an outpatient clinic for the treatment of obesity were examined from the first clinical evaluation until 1 y of diet ambulatory treatment. MEASUREMENTS: We characterised the participants at baseline on the basis of their somatic characteristics, socioeconomic status, obesity-related diseases and dietary habits. The most significant factors resulting in univariate statistical analysis (waist, body mass index (BMI), full-time job, depressive syndrome, number of obesity-related diseases, daily frequency of fruit consumption) were then examined as independent variables in direct multiple logistic regression with the dependent variable drop-out. RESULTS: The 1-y drop-out rate was 77.3%. A total of 87 patients completed the follow-up study. The noncompleter patients had slightly lower BMI and waist circumference mean values, and they were further regularly employed in full-time jobs, while the completer patients were principally pensioners and housewives. Drop-outs had a lower number of obesity related diseases and as a result were less depressed. By the logistic regression, full-time job is the best predictor of premature withdrawal (odds ratio=2.40). Age, gender, anthropometric measurements, lifestyle and dietary habits did not result as significant predictors of drop-out. CONCLUSION: The overweight and obese outpatients at higher risk of ambulatory treatment drop-out are more likely to work full hours, have less obesity-related complications and be less depressed. In our study, the full-time job condition seems to be the strongest predictor of premature withdrawal. PMID- 15545975 TI - Molecular mechanisms of action and prediction of response to oxaliplatin in colorectal cancer cells. AB - The platinum compound oxaliplatin has been shown to be an effective chemotherapeutic agent for the treatment of colorectal cancer. In this study, we investigate the molecular mechanisms of action of oxaliplatin to identify means of predicting response to this agent. Exposure of colon cancer cells to oxaliplatin resulted in G2/M arrest and apoptosis. Immunofluorescent staining demonstrated that the apoptotic cascade initiated by oxaliplatin is characterised by translocation of Bax to the mitochondria and cytochrome c release into the cytosol. Oxaliplatin treatment resulted in caspase 3 activation and oxaliplatin induced apoptosis was abrogated by inhibition of caspase activity with z-VAD-fmk, but was independent of Fas/FasL association. Targeted inactivation of Bax or p53 in HCT116 cells resulted in significantly increased resistance to oxaliplatin. However, the mutational status of p53 was unable to predict response to oxaliplatin in a panel of 30 different colorectal cancer cell lines. In contrast, the expression profile of these 30 cell lines, assessed using a 9216-sequence cDNA microarray, successfully predicted the apoptotic response to oxaliplatin. A leave-one-out cross-validation approach was used to demonstrate a significant correlation between experimentally observed and expression profile predicted apoptosis in response to clinically achievable doses of oxaliplatin (R=0.53; P=0.002). In addition, these microarray experiments identified several genes involved in control of apoptosis and DNA damage repair that were significantly correlated with response to oxaliplatin. PMID- 15545977 TI - Child temperament does not predict adolescent body composition in girls. AB - OBJECTIVES: Our previous cross-sectional analysis of MIT Growth and Development (MIT) Study girls showed that activity temperament, as assessed by a nine-item temperament questionnaire, was related to body composition and nonresting energy expenditure (NREE). In girls with lower levels of physical activity, having a high activity temperament was associated with a higher NREE. Percentage body fat was lower in girls with high vs low activity temperament. Based on these results, we hypothesized that, especially in girls with declining levels of physical activity over adolescence, high activity temperament in childhood would be protective against increased adiposity during adolescence. We tested this hypothesis with longitudinal data from the MIT study. DESIGN, SETTING, AND PARTICIPANTS: A total of 196 nonobese premenarcheal girls 8-12 y old were enrolled between 1990 and 1993. Girls were followed until 4 y postmenarche; average duration of follow-up was 7 y. MEASURES: Activity temperament was assessed at baseline by the girls' mothers with questions modified from those developed by Thomas and Chess that tapped predilection for movement. Temperament, the stylistic component of behavior, is considered relatively stable within an individual. Body composition was assessed by total body water at baseline and study completion (4 y postmenarche), and by bioelectrical impedance (BIA) annually. Physical activity was assessed annually by questionnaire, and by activity diary at baseline only. RESULTS: Child activity temperament was not associated with percentage body fat at 4 y postmenarche in multivariate regression models controlling for baseline percentage body fat, physical activity, parental obesity status, age at menarche, age at baseline, and race ethnicity. Body composition of girls with low and high activity temperaments who reported declining levels of physical activity over adolescence was not statistically significantly different at study completion. In longitudinal models of percentage body fat by BIA, high activity temperament was not associated with lower adiposity. CONCLUSION: Although high activity temperament was associated cross-sectionally with lower percentage body fat and higher NREE, we did not find evidence to support our hypothesis that high child activity temperament would be protective for increased adiposity prospectively in our cohort of girls followed over the adolescent period. PMID- 15545978 TI - Support for involvement of neuregulin 1 in schizophrenia pathophysiology. AB - Schizophrenia is a common, multigenic psychiatric disorder. Linkage studies, including a recent meta-analysis of genome scans, have repeatedly implicated chromosome 8p12-p23.1 in schizophrenia susceptibility. More recently, significant association with a candidate gene on 8p12, neuregulin 1 (NRG1), has been reported in several European and Chinese samples. We investigated NRG1 for association in schizophrenia patients of Portuguese descent to determine whether this gene is a risk factor in this population. We tested NRG1 markers and haplotypes for association in 111 parent-proband trios, 321 unrelated cases, and 242 control individuals. Associations were found with a haplotype that overlaps the risk haplotype originally reported in the Icelandic population ("Hap(ICE)"), and two haplotypes located in the 3' end of NRG1 (all P<0.05). However, association was not detected with Hap(ICE) itself. Comparison of NRG1 transcript expression in peripheral leukocytes from schizophrenia patients and unaffected siblings identified 3.8-fold higher levels of the SMDF variant in patients (P=0.039). Significant positive correlations (P<0.001) were found between SMDF and HRG-beta 2 expression and between HRG-gamma and ndf43 expression, suggesting common transcriptional regulation of NRG1 variants. In summary, our results suggest that haplotypes across NRG1 and multiple NRG1 variants are involved in schizophrenia. PMID- 15545979 TI - Prostate Cancer Charitable Trust 2004 Forum: a personal overview. PMID- 15545980 TI - Health and quality of life of persons with spinal cord lesion in Australia and Sweden. AB - STUDY DESIGN: Cross-sectional questionnaire study. OBJECTIVES: The aim of the study was to demonstrate the value of common general health-related quality of life (HRQL) instruments, readily available for cross-cultural comparisons, in persons with spinal cord lesion (SCL) treated at spinal units in Melbourne, Australia and in Gothenburg, Sweden. Another aim was to determine as to which of the independent variables were the most powerful predictors of global QL in the two SCL groups. SETTINGS: Australia and Sweden. METHODS: Two groups consisting of 89 Australian and 71 Swedish SCL persons were surveyed. The two groups were matched according to sex, age, time since injury and level of injury. The 36-item short-form health survey (SF-36) and the Spinal Cord Injuries Quality of Life-23 item questionnaire were used to cover essential generic and specific domains of health and quality of life. Clinical and sociodemographic data were collected using questionnaires. For interpretation purpose, available general population data were used. RESULTS: The most important predictors of QL perception in the Australian group were the same as for the Swedish group; mood, physical and social functioning and problems regarding injury. In total, 61% (Australian group) and 52% (Swedish group) of the variance was explained by these variables. Despite similarities of health profiles, some areas of the SF-36 differed between the two groups. CONCLUSION: The demonstrated HRQL profiles in SCL persons have shown that valid measures can provide new information of clinical value beyond the self-evident physical and practical restrictions due to injury. Illustrative comparisons, such as those presented here between Australia and Sweden, may extend our knowledge about areas where the SCL persons themselves are the logical experts, for example, maintenance of personal roles, social interaction and emotional well-being. PMID- 15545981 TI - Intrathecal drug therapy using the Codman Model 3000 Constant Flow Implantable Infusion Pumps: experience with 17 cases. AB - OBJECTIVES: The objective of this study was to evaluate the accuracy, reliability, safety, and efficacy of the Codman Model 3000 Constant Flow Implantable Infusion Pump for intrathecal baclofen delivery as a therapeutic option for the treatment of severe spasticity. The distinctive features of this pump include a raised, easily palpable septum, a safety valve protecting the bolus pathway, no programmer needed, and no battery to fail. DESIGN: A total of 17 patients with spinal cord injury, multiple sclerosis, or cerebral palsy were implanted with this pump. The accuracy of the pump and drug treatment efficacy was determined at each visit and adjustments to the dosages were made as required. All the intrathecal drug delivery system complications were reviewed. RESULTS: The expected efficacy was achieved. The accuracy of the implanted pumps ranged from 90-97% (average 94%). There were no complications due to primary pump problems. The complications reported are similar to other implantable infusion devices and include dehiscence of the suture line, pressure ulcer development, formation of seroma, inversion of the pump, baclofen overdose, and catheter failures. CONCLUSION: The Codman Model 3000 Constant Flow Implantable Infusion Pump is an accurate, reliable, and convenient option for patients needing intrathecal baclofen therapy, with complications similar to other available pumps. PMID- 15545982 TI - Targeting oncogenes. PMID- 15545983 TI - Stem cells: science, policy, and ethics. AB - Human embryonic stem cells offer the promise of a new regenerative medicine in which damaged adult cells can be replaced with new cells. Research is needed to determine the most viable stem cell lines and reliable ways to promote the differentiation of pluripotent stem cells into specific cell types (neurons, muscle cells, etc). To create new cell lines, it is necessary to destroy preimplantation blastocysts. This has led to an intense debate that threatens to limit embryonic stem cell research. The profound ethical issues raised call for informed, dispassionate debate. PMID- 15545984 TI - IL-10-producing and naturally occurring CD4+ Tregs: limiting collateral damage. AB - Effective immune responses against pathogens are sometimes accompanied by strong inflammatory reactions. To minimize damage to self, the activation of the immune system also triggers anti-inflammatory circuits. Both inflammatory and anti inflammatory reactions are normal components of the same immune response, which coordinately fight infections while preventing immune pathology. IL-10 is an important suppressive cytokine, produced by a large number of immune cells in addition to the antigen-driven IL-10-producing regulatory and the naturally occurring suppressor CD4+ T cells, which is a key player in anti-inflammatory immune responses. However, additional mechanisms have evolved to ensure that pathogen eradication is achieved with minimum damage to the host. Here we discuss those mechanisms that operate to regulate effector immune responses. PMID- 15545985 TI - Going both ways: immune regulation via CD1d-dependent NKT cells. AB - NKT cells are a unique T lymphocyte sublineage that has been implicated in the regulation of immune responses associated with a broad range of diseases, including autoimmunity, infectious diseases, and cancer. In stark contrast to both conventional T lymphocytes and other types of Tregs, NKT cells are reactive to the nonclassical class I antigen-presenting molecule CD1d, and they recognize glycolipid antigens rather than peptides. Moreover, they can either up- or downregulate immune responses by promoting the secretion of Th1, Th2, or immune regulatory cytokines. This review will explore the diverse influences of these cells in various disease models, their ability to suppress or enhance immunity, and the potential for manipulating these cells as a novel form of immunotherapy. PMID- 15545987 TI - Tregs and transplantation tolerance. AB - The induction and maintenance of immune tolerance to transplanted tissues constitute an active process involving multiple mechanisms that work cooperatively to prevent graft rejection. These mechanisms are similar to inherent tolerance toward self antigens and have a requirement for active immunoregulation, largely T cell mediated, that promotes specific unresponsiveness to donor alloantigens. This review outlines our current understanding of the Treg subsets that contribute to allotolerance and the mechanisms by which these cells exert their effects as well as their potential for therapy. PMID- 15545986 TI - Tregs and allergic disease. AB - Allergic diseases such as asthma, rhinitis, and eczema are increasing in prevalence and affect up to 15% of populations in Westernized countries. The description of Tregs as T cells that prevent development of autoimmune disease led to considerable interest in whether these Tregs were also normally involved in prevention of sensitization to allergens and whether it might be possible to manipulate Tregs for the therapy of allergic disease. Current data suggest that Th2 responses to allergens are normally suppressed by both CD4+CD25+ Tregs and IL 10 Tregs. Furthermore, suppression by these subsets is decreased in allergic individuals. In animal models, Tregs could be induced by high- or low-dose inhaled antigen, and prior induction of such Tregs prevented subsequent development of allergen sensitization and airway inflammation in inhaled challenge models. For many years, allergen-injection immunotherapy has been used for the therapy of allergic disease, and this treatment may induce IL-10 Tregs, leading to both suppression of Th2 responses and a switch from IgE to IgG4 antibody production. Improvements in allergen immunotherapy, such as peptide therapy, and greater understanding of the biology of Tregs hold great promise for the treatment and prevention of allergic disease. PMID- 15545989 TI - Piecing together the puzzle of cutaneous mosaicism. AB - Autosomal dominant disorders of the skin may present in a pattern following the lines of embryologic development of the ectoderm. In these cases, the surrounding skin is normal, and molecular studies have shown that the causative mutation is confined to the affected ectodermal tissue (type 1 mosaicism). Rarely, an individual shows skin lesions that follow the pattern of type 1 mosaicism, but the rest of the skin shows a milder form of the disorder (type 2 mosaicism). A new study provides the molecular basis for type 2 mosaicism. PMID- 15545988 TI - RIP-ed and ready to dance: new mechanisms for polycystin-1 signaling. AB - Polycystin-1, the protein encoded by the principal gene involved in autosomal dominant polycystic kidney disease, has been implicated in extracellular sensing as well as in cell-cell and cell-matrix interactions. However, the precise mechanisms involved in polycystin-1 signaling are not well defined. A report in this issue of the JCI demonstrates that the C-terminal tail of polycystin-1 is cleaved from the membrane through regulated intramembrane proteolysis (RIP) and that this domain translocates to the nucleus, where it activates the AP-1 transcription pathway. This translocation appears to be modulated by polycystin 2, with which polycystin-1 is thought to interact. These findings provide what we believe to be the first evidence that polycystin-1 can signal directly to the nucleus and that polycystin-1-polycystin-2 interactions do not require colocalization of these proteins in the same membrane compartment. PMID- 15545990 TI - The multiple causes of human SCID. AB - SCID, a syndrome characterized by the absence of T cells and adaptive immunity, can result from mutations in multiple genes that encode components of the immune system. Three such components are cytokine receptor chains or signaling molecules, five are needed for antigen receptor development, one is adenosine deaminase--a purine salvage pathway enzyme, and the last is a phosphatase, CD45. In this issue of the JCI, a report describes how complete deficiency of the CD3epsilon chain of the T cell antigen receptor/CD3 complex causes human SCID. PMID- 15545991 TI - What determines glomerular capillary permeability? AB - There have been exciting recent advances in our understanding of the structural and molecular biology of the glomerular slit diaphragm, as described in a report in this issue of the JCI. These findings, combined with data on the permeability of the basement membrane and evidence that the endothelium may be a more important barrier than often supposed, are allowing a clearer understanding to emerge of how the 3 parts of the glomerular capillary wall jointly determine its functional properties. PMID- 15545992 TI - Genes and pathophysiology of type 2 diabetes: more than just the Randle cycle all over again. AB - The Randle cycle, which has been invoked to explain the reciprocal relationship between fatty acid oxidation and glucose oxidation, has long been implicated as a potential mechanism for hyperglycemia and type 2 diabetes mellitus (T2DM). Now genetic, functional genomic, and transgenic approaches have identified PPARgamma coactivators (PGC-1alpha and PGC-1beta) as key regulators of mitochondrial number and function. They regulate adaptive thermogenesis as well as glucose and fat oxidation in muscle and fat tissue, gluconeogenesis in liver, and even glucose regulated insulin secretion in beta cells. PGC-1alpha and PGC-1beta mRNA levels and the mitochondrial genes they regulate are decreased in muscle of people with prediabetes and T2DM. A new report indicates that PGC-1alpha and PGC-1beta mRNA levels decrease with age in individuals with a genetic variant in PGC-1alpha, and these decreases correlate with alterations in whole-body glucose and fatty acid oxidation. These findings provide insights into how aging modifies genetic susceptibility to alterations in oxidative phosphorylation and T2DM. PMID- 15545993 TI - An uncleavable form of pro-scatter factor suppresses tumor growth and dissemination in mice. AB - Scatter factor (SF), also known as hepatocyte growth factor, is ubiquitously present in the extracellular matrix of tissues in the form of an inactive precursor (pro-SF). In order to acquire biological activity, pro-SF must be cleaved by specific proteases present on the cell surface. The mature form of SF controls invasive cues in both physiological and pathological processes through activation of its receptor, the Met tyrosine kinase. By substituting a single amino acid in the proteolytic site, we engineered an unprocessable form of pro-SF (uncleavable SF). Using lentivirus vector technology, we achieved local or systemic delivery of uncleavable SF in mice. We provide evidence that (a) uncleavable SF inhibits both protease-mediated pro-SF conversion and active SF induced Met activation; (b) local expression of uncleavable SF in tumors suppresses tumor growth, impairs tumor angiogenesis, and prevents metastatic dissemination; and (c) systemic expression of uncleavable SF dramatically inhibits the growth of transplanted tumors and abolishes the formation of spontaneous metastases without perturbing vital physiological functions. These data show that proteolytic activation of pro-SF is a limiting step in tumor progression, thus suggesting a new strategy for the treatment or prevention of the malignant conversion of neoplastic lesions. PMID- 15545994 TI - Mechanical stimuli induce cleavage and nuclear translocation of the polycystin-1 C terminus. AB - Polycystin-1, which is encoded by a gene that is mutated in autosomal dominant polycystic kidney disease (ADPKD), is involved in cell-matrix interactions as well as in ciliary signaling. The precise mechanisms by which it functions, however, remain unclear. Here we find that polycystin-1 undergoes a proteolytic cleavage that releases its C-terminal tail (CTT), which enters the nucleus and initiates signaling processes. The cleavage occurs in vivo in association with alterations in mechanical stimuli. Polycystin-2, the product of the second gene mutated in ADPKD, modulates the signaling properties of the polycystin-1 CTT. These data reveal a novel pathway by which polycystin-1 transmits messages directly to the nucleus. PMID- 15545995 TI - A role for proteinase-activated receptor-1 in inflammatory bowel diseases. AB - Proteinase-activated receptor-1 (PAR1), a G protein-coupled receptor activated by thrombin, is highly expressed in different cell types of the gastrointestinal tract. The activity of thrombin and of other proteinases is significantly increased in the colon of inflammatory bowel disease (IBD) patients. Since PAR1 activation in tissues other than the gut provoked inflammation, we hypothesized that PAR1 activation in the colon is involved in the pathogenesis of IBD. Here, we demonstrate that PAR1 is overexpressed in the colon of IBD patients. In mice, intracolonic administration of PAR1 agonists led to an inflammatory reaction characterized by edema and granulocyte infiltration. This PAR1 activation-induced inflammation was dependent on B and T lymphocytes. Moreover, PAR1 activation exacerbated and prolonged inflammation in a mouse model of IBD induced by the intracolonic administration of trinitrobenzene sulfonic acid (TNBS), while PAR1 antagonism significantly decreased the mortality and severity of colonic inflammation induced by TNBS and dextran sodium sulfate. In these 2 models, colitis development was strongly attenuated by PAR1 deficiency. Taken together, these results imply an important role for PAR1 in the pathogenesis of experimental colitis, supporting the notion that PAR1 inhibition may be beneficial in the context of IBD and possibly in other chronic intestinal inflammatory disorders. PMID- 15545996 TI - Loss of alpha-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates beta-thalassemia. AB - Hemoglobin (Hb) A production during red blood cell development is coordinated to minimize the deleterious effects of free alpha- and beta-Hb subunits, which are unstable and cytotoxic. The alpha-Hb-stabilizing protein (AHSP) is an erythroid protein that specifically binds alpha-Hb and prevents its precipitation in vitro, which suggests that it may function to limit free alpha-Hb toxicities in vivo. We investigated this possibility through gene ablation and biochemical studies. AHSP(-/-) erythrocytes contained hemoglobin precipitates and were short-lived. In hematopoietic tissues, erythroid precursors were elevated in number but exhibited increased apoptosis. Consistent with unstable alpha-Hb, AHSP(-/-) erythrocytes contained increased ROS and evidence of oxidative damage. Moreover, purified recombinant AHSP inhibited ROS production by alpha-Hb in solution. Finally, loss of AHSP worsened the phenotype of beta-thalassemia, a common inherited anemia characterized by excess free alpha-Hb. Together, the data support a model in which AHSP binds alpha-Hb transiently to stabilize its conformation and render it biochemically inert prior to Hb A assembly. This function is essential for normal erythropoiesis and, to a greater extent, in beta-thalassemia. Our findings raise the possibility that altered AHSP expression levels could modulate the severity of beta-thalassemia in humans. PMID- 15545997 TI - Allelic loss underlies type 2 segmental Hailey-Hailey disease, providing molecular confirmation of a novel genetic concept. AB - Hailey-Hailey disease (HHD) is an autosomal dominant trait characterized by erythematous and oozing skin lesions preponderantly involving the body folds. In the present unusual case, however, unilateral segmental areas along the lines of Blaschko showing a rather severe involvement were superimposed on the ordinary symmetrical phenotype. Based on this observation and similar forms of mosaicism as reported in other autosomal dominant skin disorders, we postulated that in such cases, 2 different types of segmental involvement can be distinguished. Accordingly, the linear lesions as noted in the present case would exemplify type 2 segmental HHD. In the heterozygous embryo, loss of heterozygosity occurring at an early developmental stage would have given rise to pronounced linear lesions reflecting homozygosity or hemizygosity for the mutation. By analyzing DNA and RNA derived from blood and skin samples as well as keratinocytes of the index patient with various molecular techniques including RT-PCR, real-time PCR, and microsatellite analysis, we found a consistent loss of the paternal wild-type allele in more severely affected segmental skin regions, confirming this hypothesis for the first time, to our knowledge, at the molecular and cellular level. PMID- 15545998 TI - Nephrin strands contribute to a porous slit diaphragm scaffold as revealed by electron tomography. AB - Nephrin is a key functional component of the slit diaphragm, the structurally unresolved molecular filter in renal glomerular capillaries. Abnormal nephrin or its absence results in severe proteinuria and loss of the slit diaphragm. The diaphragm is a thin extracellular membrane spanning the approximately 40-nm-wide filtration slit between podocyte foot processes covering the capillary surface. Using electron tomography, we show that the slit diaphragm comprises a network of winding molecular strands with pores the same size as or smaller than albumin molecules, as demonstrated in humans, rats, and mice. In the network, which is occasionally stratified, immunogold-nephrin antibodies labeled individually detectable globular cross strands, about 35 nm in length, lining the lateral elongated pores. The cross strands, emanating from both sides of the slit, contacted at the slit center but had free distal endings. Shorter strands associated with the cross strands were observed at their base. Immunolabeling of recombinant nephrin molecules on transfected cells and in vitrified solution corroborated the findings in kidney. Nephrin-deficient proteinuric patients with Finnish-type congenital nephrosis and nephrin-knockout mice had only narrow filtration slits that lacked the slit diaphragm network and the 35-nm-long strands but contained shorter molecular structures. The results suggest the direct involvement of nephrin molecules in constituting the macromolecule retaining slit diaphragm and its pores. PMID- 15545999 TI - Defining the pathogenic involvement of desmoglein 4 in pemphigus and staphylococcal scalded skin syndrome. AB - Desmogleins (Dsgs), cadherin-type cell adhesion molecules, are targeted in skin blistering diseases such as pemphigus and staphylococcal scalded skin syndrome (SSSS). The role of Dsg4, a new isoform, was investigated in these diseases. Dsg4 was recognized by 30 (77%) of 39 pemphigus sera containing anti-Dsg1 IgG but not by 16 pemphigus sera containing no anti-Dsg1 IgG or by 34 normal control sera. The Dsg4 immunoreactivity of these sera was abolished by removal of anti-Dsg1 IgG. Conversely, the removal of anti-Dsg4 IgG from pemphigus sera reduced the immunoreactivity against Dsg1 only 13.8% +/- 8.8% (n = 23) and did not affect its ability to induce blisters in neonatal mice. IgG that was affinity-purified on Dsg4 recognized Dsg1 but failed to induce blisters, while IgG purified on Dsg1 from the same pemphigus foliaceus sera induced blisters. Thus, pemphigus sera show Dsg4 reactivity due to cross-reactivity of a subset of anti-Dsg1 IgG, and the Dsg4/Dsg1-cross-reacting IgG has no demonstrable pathogenic effect. In addition, Dsg4 was not cleaved by exfoliative toxins that induce blisters in SSSS. These findings suggest that Dsg4 may play a role other than adhesion and that the cross-reactivity of desmoglein autoantibodies should be factored into the framework of future studies of autoimmune mechanisms in pemphigus. PMID- 15546000 TI - Foxo1 mediates insulin action on apoC-III and triglyceride metabolism. AB - The apolipoprotein apoC-III plays an important role in plasma triglyceride metabolism. It is predominantly produced in liver, and its hepatic expression is inhibited by insulin. To elucidate the inhibitory mechanism of insulin in apoC III expression, we delivered forkhead box O1 (Foxo1) cDNA to hepatocytes by adenovirus-mediated gene transfer. Foxo1 stimulated hepatic apoC-III expression and correlated with the ability of Foxo1 to bind to its consensus site in the apoC-III promoter. Deletion or mutation of the Foxo1 binding site abolished insulin response and Foxo1-mediated stimulation. Likewise, Foxo1 also mediated insulin action on intestinal apoC-III expression in enterocytes. Furthermore, elevated Foxo1 production in liver augmented hepatic apoC-III expression, resulting in increased plasma triglyceride levels and impaired fat tolerance in mice. Transgenic mice expressing a constitutively active Foxo1 allele exhibited hypertriglyceridemia. Moreover, we show that hepatic Foxo1 expression becomes deregulated as a result of insulin deficiency or insulin resistance, culminating in significantly elevated Foxo1 production, along with its skewed nuclear distribution, in livers of diabetic NOD or db/db mice. While loss of insulin response is associated with unrestrained apoC-III production and impaired triglyceride metabolism, these data suggest that Foxo1 provides a molecular link between insulin deficiency or resistance and aberrant apoC-III production in the pathogenesis of diabetic hypertriglyceridemia. PMID- 15546001 TI - Disruption of either the Nfkb1 or the Bcl3 gene inhibits skeletal muscle atrophy. AB - The intracellular signals that mediate skeletal muscle protein loss and functional deficits due to muscular disuse are just beginning to be elucidated. Previously we showed that the activity of an NF-kappaB-dependent reporter gene was markedly increased in unloaded muscles, and p50 and Bcl-3 proteins were implicated in this induction. In the present study, mice with a knockout of the p105/p50 (Nfkb1) gene are shown to be resistant to the decrease in soleus fiber cross-sectional area that results from 10 days of hindlimb unloading. Furthermore, the marked unloading-induced activation of the NF-kappaB reporter gene in soleus muscles from WT mice was completely abolished in soleus muscles from Nfkb1 knockout mice. Knockout of the B cell lymphoma 3 (Bcl3) gene also showed an inhibition of fiber atrophy and an abolition of NF-kappaB reporter activity. With unloading, fast fibers from WT mice atrophied to a greater extent than slow fibers. Resistance to atrophy in both strains of knockout mice was demonstrated clearly in fast fibers, while slow fibers from only the Bcl3(-/-) mice showed atrophy inhibition. The slow-to-fast shift in myosin isoform expression due to unloading was also abolished in both Nfkb1 and Bcl3 knockout mice. Like the soleus muscles, plantaris muscles from Nfkb1(-/-) and Bcl3(-/-) mice also showed inhibition of atrophy with unloading. Thus both the Nfkb1 and the Bcl3 genes are necessary for unloading-induced atrophy and the associated phenotype transition. PMID- 15546002 TI - Severe combined immunodeficiency caused by deficiency in either the delta or the epsilon subunit of CD3. AB - We investigated the molecular mechanism underlying a severe combined immunodeficiency characterized by the selective and complete absence of T cells. The condition was found in 5 patients and 2 fetuses from 3 consanguineous families. Linkage analysis performed on the 3 families revealed that the patients were carrying homozygous haplotypes within the 11q23 region, in which the genes encoding the gamma, delta, and epsilon subunits of CD3 are located. Patients and affected fetuses from 2 families were homozygous for a mutation in the CD3D gene, and patients from the third family were homozygous for a mutation in the CD3E gene. The thymus from a CD3delta-deficient fetus was analyzed and revealed that T cell differentiation was blocked at entry into the double positive (CD4+CD8+) stage with the accumulation of intermediate CD4-single positive cells. This indicates that CD3delta plays an essential role in promoting progression of early thymocytes toward double-positive stage. Altogether, these findings extend the known molecular mechanisms underlying severe combined immunodeficiency to a new deficiency, i.e., CD3epsilon deficiency, and emphasize the essential roles played by the CD3epsilon and CD3delta subunits in human thymocyte development, since these subunits associate with both the pre-TCR and the TCR. PMID- 15546003 TI - Multiple environmental and genetic factors influence skeletal muscle PGC-1alpha and PGC-1beta gene expression in twins. AB - Genetic and environmental factors contribute to age-dependent susceptibility to type 2 diabetes. Recent studies have reported reduced expression of PPARgamma coactivator 1alpha (PGC-1alpha) and PGC-1beta genes in skeletal muscle from type 2 diabetic patients, but it is not known whether this is an inherited or acquired defect. To address this question we studied expression of these genes in muscle biopsies obtained from young and elderly dizygotic and monozygotic twins without known diabetes before and after insulin stimulation and related the expression to a Gly482Ser variant in the PGC-1alpha gene. Insulin increased and aging reduced skeletal muscle PGC-1alpha and PGC-1beta mRNA levels. This age-dependent decrease in muscle gene expression was partially heritable and influenced by the PGC 1alpha Gly482Ser polymorphism. In addition, sex, birth weight, and aerobic capacity influenced expression of PGC-1alpha in a complex fashion. Whereas expression of PGC-1alpha in muscle was positively related to insulin-stimulated glucose uptake and oxidation, PGC-1beta expression was positively related to fat oxidation and nonoxidative glucose metabolism. We conclude that skeletal muscle PGC-1alpha and PGC-1beta expression are stimulated by insulin and reduced by aging. The data also suggest different regulatory functions for PGC-1alpha and PGC-1beta on glucose and fat oxidation in muscle cells. The finding that the age dependent decrease in the expression of these key genes regulating oxidative phosphorylation is under genetic control could provide an explanation by which an environmental trigger (age) modifies genetic susceptibility to type 2 diabetes. PMID- 15546008 TI - European orthodontic specialists in 2002. AB - After clarifying the role and significance of the European Federations of Orthodontic Specialists Associations (EFOSA), the results of a recent survey examining the situation of the Speciality of Orthodontics in Europe are presented. Among the many items included are the recognition and availability of orthodontic specialists, their training, and working conditions, the fees charged, and the present insurance and refunding systems. PMID- 15546009 TI - Orthodontic therapy and third party in Europe. AB - This paper is an attempt to individuate some principles and guidelines apt to regulate the relationship between orthodontists and financing third parties, applicable to most western European Countries. The concepts of orthodontic treatment need, orthodontic treatment request and orthodontic screening are discussed, alongside with a short overview of some of the most common indexes to assess the severity of the malocclusion and/or the treatment priority. The screening method introduced by the Danish Ministry of Health is presented; its importance lies in the fact that for the first time a direct correlation between health risk and individual malocclusions is recognized and assessed. In the discussion, it is stressed how the screening system tightly depends on the chosen general model for orthodontic care. Different models of orthodontic care organization as presently used in many European countries are presented and shortly discussed; among these, the Norwegian model is described more in details, because of its simplicity. Eventually, some guidelines considered necessary in order to achieve satisfactory standards of quality and efficiency are presented and discussed. PMID- 15546004 TI - TGF-beta-dependent pathogenesis of mitral valve prolapse in a mouse model of Marfan syndrome. AB - Mitral valve prolapse (MVP) is a common human phenotype, yet little is known about the pathogenesis of this condition. MVP can occur in the context of genetic syndromes, including Marfan syndrome (MFS), an autosomal-dominant connective tissue disorder caused by mutations in fibrillin-1. Fibrillin-1 contributes to the regulated activation of the cytokine TGF-beta, and enhanced signaling is a consequence of fibrillin-1 deficiency. We thus hypothesized that increased TGF beta signaling may contribute to the multisystem pathogenesis of MFS, including the development of myxomatous changes of the atrioventricular valves. Mitral valves from fibrillin-1-deficient mice exhibited postnatally acquired alterations in architecture that correlated both temporally and spatially with increased cell proliferation, decreased apoptosis, and excess TGF-beta activation and signaling. In addition, TGF-beta antagonism in vivo rescued the valve phenotype, suggesting a cause and effect relationship. Expression analyses identified increased expression of numerous TGF-beta-related genes that regulate cell proliferation and survival and plausibly contribute to myxomatous valve disease. These studies validate a novel, genetically engineered murine model of myxomatous changes of the mitral valve and provide critical insight into the pathogenetic mechanism of such changes in MFS and perhaps more common nonsyndromic variants of mitral valve disease. PMID- 15546010 TI - Rapid palatal expansion and pharyngeal space. Cephalometric evaluation. AB - OBJECTIVES: the aim of this study was to evaluate cephalometrically the upper respiratory airway dimensions before and after rapid maxillary expansion. MATERIALS AND METHODS: rapid palatal expansion was performed on 24 5-9 year old children with adenotonsillar hypertrophy determined radiographically by means of lateral cephalographs. Also, the subjects were questioned concerning their ability to breath through their noses. RESULTS: there was an increase in upper respiratory width that often coincided with a reported improvement in nasal respiration. Adenoid volume was not reduced. CONCLUSIONS: the increase in pharyngeal space and improvement in nasal breathing resulted from an increase in pharyngeal lumen enlargement rather than a reduction in the volume of the adenoid tissue. PMID- 15546011 TI - Lower incisor extraction in Class I and Class II malocclusions: case reports. AB - After a brief discussion of the indications/contraindications of extracting lower incisors as part of orthodontic treatment, five case reports are presented. These cases highlight that extracting lower incisors can be an accepted orthodontic option, not only in compromised class III treatments, but also in Class I and Class II malocclusions. PMID- 15546012 TI - Dental arch analysis system. AB - The purpose of our research is to describe a computerized method, which enables orthodontists and researchers to analyze variation of dental arch form and dimension. The analysis system is composed of two independent parts: the database, where the images of scanned dental casts are stored, and the software. The operator uses the software to identify some landmarks on dental cast images. Corresponding distances are calculated (interincisive, intercanine and intermolar widths and the curve axis). Then the software algorithms calculate and draw the curves passing trough the selected landmarks (conics, catenary, cubic spline and polynomial curves). In the chosen curves the arch length is measured. The dental cast data, recorded at different times of the subject's life or in different steps of orthodontic treatment, are compared in order to evaluate the change in dimension (arch length and width) and form (shape). The statistical analysis of the data evaluates the variation in form and in dimension separately. The shape change is defined by Euclidean Distance Matrix Analysis (EDMA) and Bootstrap analysis. The computerized method allows orthodontists and researches to evaluate the variation in dimension and form of the dental arch in un-treated subjects and or orthodontic patients over time. After the analysis of a large sample of patients or subjects, information concerning changes in dental arch dimension and form can be added to previous studies of other authors, distinguishing variation during and after orthodontic treatment, during growth or aging. PMID- 15546013 TI - Clinical applications of the Mini-Screw-Anchorage-System (M.A.S.) in the maxillary alveolar bone. AB - AIMS: anchorage control with self-tapping screws has become an important part of the clinical management of the orthodontic patients. Mechanical resistance and sites of insertion of miniscrews as orthodontic anchorage are critical to ensure successful outcomes. Aim of this clinical study was threefold: 1) to measure the mechanical resistance of the M.A.S., 2) to evaluate if the alveolar areas usually selected for mini-screws placement are adequate, 3) to illustrate the most frequent clinical application on the maxillary alveolar bone. METHODS: two methods were chosen to test these screws mechanically, representing two potential modes of failure during insertion or removal: torsional strenght and bending strenght. Three-dimension images of fifty maxillas have been retrieved from a group of 200 patients, age range between 20 and 40 years with a new type of tomogram called Newtom System. For each area mesio-distal and labio-lingual measurements from four horizontal cuts made at 2-5-8-11 mm below the bone-crest have been evaluated. RESULTS: the mean value of resistance to breakage in torsion is of 48.7 N.cm (around 5 Kg) for the miniscrew of 1.5 diameter, while the mean value of resistance to breakage in torsion is of 23.4 N.cm (around 2 Kg) for the miniscrew of 1.3 diameter.. The mean value of resistance to breakage in flexion is of 120.4 N (around 12 Kg) for the miniscrew of 1.5 diameter, while the mean value of resistance to the flexion is of 63.7 N (around 6 Kg) for the miniscrew of 1.3 diameter. On the maxillary alveolar bone the highest amount of bone was in mesio-distal dimension between 6 and 5 on the palatal side (minimum 1.9 mm at -11 mm cut; maximum 5.5 mm at -5 mm cut). The smallest amount of bone was in the tuber (minimum 0.2 mm; maximum 1.3 mm). Examination of the labio-palatal dimension demonstrated similar high thickness between 5-6 and 6-7 (minimum 3.7 mm at -11 mm cut; maximum 13.2 mm at -2 mm cut). The smallest amount of bone was recorded on the tuber (minimum 0.6 mm; maximum 4.1 mm). The following clinical applications are described: Closure of the extractions space, Symmetric intrusion of the incisors, Correction of the cant of the plane of occlusion and of the dental midline, Molar intrusion of one or two teeth, Molar distalization with the Distal Jet and miniscrews, Molar mesialization, Intermaxillary anchorage. CONCLUSIONS: the mechanical resistance of the miniscrews M.A.S. is suitable for their use in orthodontics. The best anatomical zones for their implantation are the interradicular spaces mesial to the first maxillary molars. From our experience to date, the miniscrews are a reliable and convenient system for skeletal anchorage when compared with other more invasive osseo-integrated systems. PMID- 15546014 TI - The use of titanium miniscrews for molar protraction in extraction treatment. AB - Orthodontic space closure in the mandibular arch by protraction of the mandibular second molars, after the extraction of first molars, may sometimes result in loss of incisor anchorage when using conventional orthodontic procedures. The introduction of miniscrews for immediate loading as orthodontic anchorage, has enlarged treatment possibilities. The authors illustrate their clinical experience in an adult patient treated with the extraction of mandibular first molars and the protraction of second and third molars into the extraction sites. Anchorage control was achieved with the surgical insertion of titanium miniscrews for immediate loading in the cortical bone distal to second bicuspids. Space closure was achieved by means of sliding mechanics according to Bidimensional Technique. The position of lower incisors was maintained preventing any detrimental facial effect. PMID- 15546015 TI - Digital treatment objectives: procedure and clinical application. AB - OBJECTIVES: the introduction of digital study models in the orthodontic practice changed the traditional stone gipsotec to a virtual gipsotec and the orthodontist has the possibility to see, immediately on the computer, this basic analysis with a great reduction of space and time. The aim of this study is to extend the use of digital models to the occlusal virtual set-up in 3D and have the orthodontist simulate and visualize the resulting occlusion from a suggested treatment plan for the malocclusion. METHODS AND MATERIALS: a case is presented where the treatment plan involvers extractions. The malocclusion is a Class II division 1 in the permanent dentition where an extraction treatment of 1.4, 2.4, 3.5 et 4.5 is planned: the final result shows the new occlusal relationship with proper anchorage management of the extraction spaces. From the primary silicon impression, a digital model is obtained followed by the virtual set up according the prescription of the treatment plan. More digital treatment objectives can be performed for the same malocclusion from the primary digital models, according to different treatment plans and the resulting occlusion visualized. RESULTS: the basic principle for the virtual occlusal set-up is the straight-wire system with the final position of the teeth resulting both from the bracket prescription and the final rectangular wire in a full slot engagement. CONCLUSIONS: the resulting dental arches show a correct arch form, arch coordination and a proper dental intercuspation. The results coming from different virtual simulations are analyzed; all these simulations need only one pair of impressions. Limits of this virtual method are that it can be used only in full permanent dentition and for cases of dento-alveolar or dento-dental discrepancy. PMID- 15546016 TI - Italian Board of Orthodontics. Case Nr. 1: early treatment. AB - The patient was a 9 year 3 month old female with a normodivergent skeleto-dental Class II malocclusion in the mixed dentition characterized by maxillary incisors which were both protrusive and spaced. There was by a deep Curve of Spee and a deep bite with palatal impingement. Facially, the profile was convex when compared to the esthetic line. The nasolabial angle was normal and the face was symmetrical (Figs. 1-8, Tab. I). The functional movements and T.M.J analysis were within normal limits. The periodontum and hygeine were good. Because of the patient's age, a 2 phase non extraction treatment approach was planned. In the first phase, the intent was to reduce the overbite and overjet for funtional reasons, reducing the possibility of trauma to the maxillary incisors, and control the Class II skeltodental relationships. In the second phase, Class I relationships would be obtained with a normal overjet and overbite and an improved profile. However, in reality, treatment became a 1 phase treatment because there was no interruption between the first and second phase of treatment. PMID- 15546017 TI - Commentary on: Mobley AJ, Lam YW, Lau KM, Pais VM, Lesperance JO, Steadman B, et al. Monitoring the serological Proteome Colon, the latest modality in prostate cancer detection. J Urol 2004; 172: 331-7. PMID- 15546019 TI - Quantitative (stereological) study of incomplete spermatogenic suppression induced by testosterone undecanoate injection in rats. AB - AIM: To evaluate the key lesions in spermatogenesis suppressed partially by testosterone undecanoate (TU) treatment. METHODS: Adult male SD rats were treated with vehicle or TU (19 mg/kg) injection (i.m.) every 15 days for 130 days. The numbers of all types of cells (nuclei) in the seminiferous tubules and the interstitial tissue were estimated using a contemporary stereological tool, the optical disector. RESULTS: In response to TU treatment, the numbers of non-type B spermatogonia, type B spermatogonia and late elongated spermatids per testis were reduced to 51 %, 66 % and 14 % of the controls, respectively. The conversion ratios from type B spermatogonia to early spermatocytes and pachytene spermatocytes were not significantly affected and the ratios to the later germ cell types fell to 51 % - 65 % of the controls. Less than 1.0 % of immature round spermatids were seen sloughing into the tubule lumen, 4.0 % of elongated spermatids retained in the seminiferous epithelium, and about half of the elongated spermatid nuclei appreciably malformed. Leydig cells were atrophied but their number and the peritubular myoid cell number per testis were unchanged. CONCLUSION: Double inhibition of spermatogenesis (i.e. inhibition at spermiation and spermatogonial conversion to type B spermatogonia), a scenario seen in the monkey and human following gonadotrophin withdrawal, was not sufficiently effective for a complete spermatogenic suppression in the rat after TU treatment, probably due to ineffective inhibition of the Leydig cell population and therefore the intra-testicular testosterone levels. PMID- 15546018 TI - Combination of genistein with ionizing radiation on androgen-independent prostate cancer cells. AB - AIM: To study the effect of the combined use of genistein and ionizing radiation (IR) on prostate DU145 cancer cells. METHODS: DU145, an androgen-independent human prostate cancer cell line, was used in the experiment. Clonogenic assay was used to compare the survival of DU145 cells after treatments with genistein alone and in combination with graded IR. Apoptosis was assayed by DNA ladder and TUNEL stain. Cell cycle alterations were observed by flow cytometry and related protein expressions by immunoblotting. RESULTS: Clonogenic assay demonstrated that genistein, even at low to medium concentrations, enhanced the radiosensitivity of DU145 cells. Twenty-four hours after treatment with IR and/or genistein, apoptosis was mainly seen with genistein at high concentrations and was minimally related to IR. At 72 h, apoptosis also occurred in treatment with lower concentration of genistein, especially when combined with IR. While both IR and genistein led to G2/M cell cycle arrest, combination of them further increased the DU145 cells at G2/M phase. This G2/M arrest was largely maintained at 72 h, accompanied by increasing apoptosis and hyperdiploid cell population. Cell-cycle related protein analysis disclosed biphasic changes in cyclin B1 and less dramatically cdc-2, but stably elevated p21 cip1 levels with increasing genistein concentrations. CONCLUSION: Genistein enhanced the radiosensitivity of DU145 prostate cancer cells. The mechanisms might be involved in the increased apoptosis, prolonged cell cycle arrest and impaired damage repair. PMID- 15546020 TI - A preliminary study on the rheological properties of human ejaculate and changes during liquefaction. AB - AIM: To study the changes in rheological properties, namely the parameters of the hysteresis loops and yield stress versus time for human semen after ejaculation. METHODS: Ejaculates were obtained from volunteers and immediately put into the test cup of a Brookfield Programmable DV-11 Rheometer, by which the hysteresis loops and yield stress were determined. RESULTS: (1) Yield stress values dropped down from more than 3000 mPa to 60 mPa in about 5 minutes after ejaculation; (2) The shape of the hysteresis loops of shear stress versus shear rate was changed from the counter-clockwise direction, that enclosed a large area, into the clockwise direction, that enclosed a very small area. CONCLUSION: Human ejaculate originally possesses semi-solid or visco-elastic body behavior and in 5 minutes after liquefaction, it becomes a thixotropic fluid or shearing thinning fluid with very low viscosity. PMID- 15546021 TI - Male infertility: risk factors in Mongolian men. AB - AIM: To determine the most common risk factors of male infertility in Mongolian men attending an infertility clinic. METHODS: A prospective, case-control study was conducted in which 430 men were enrolled. All the men had sought their first infertility evaluation between 1998-2002 in the State Research Center on Maternal Child Health, Ulaanbaatar, Mongolia. They were divided into two groups depending on the results of their semen analysis: 191 with abnormal semen and 239 with normal semen profile. Univariate and multivariate analyses were performed to determine any association between risk factors and semen abnormality. RESULTS: Logistic regression analysis demonstrated that the testicular volume, a history of sexually transmitted infections (STI), epididymitis and testicular damage all have statistically significant associations with semen abnormality, when controlled for multiple risk factors. Adjusted odds ratios of 3.4 for mumps orchitis, 2.3 for other orchitis and 3.9 for testicular injury were found. Gonorrhoea, the most commonly reported STIs in this study, gave an adjusted odds ratio of 1.0 for having one or more sperm abnormality. An adjusted odds ratio for subjects with a history of other STIs was 2.7. However, as a predictor of azoospermia, STIs had very high odds ratio, being 5.6 in patients with gonorrhoea and 7.6 in patients with other STIs. CONCLUSION: A history of pathology involving testicular damage appeared to have the strongest impact on male infertility in Mongolia. STIs have less impact on semen quality except when complicated by orchitis, epididymitis and vasal obstruction. PMID- 15546023 TI - Abrogation of heat-shock protein (HSP)70 expression induced cell growth inhibition and apoptosis in human androgen-independent prostate cancer cell line PC-3m. AB - AIM: To investigate the effect of abrogating heat shock protein (HSP) 70 expression by antisense HSP70 oligonucleotides treatment on human androgen independent prostate cancer cell line PC-3m growth. METHODS: PC-3m cells were treated with 0-16 mmol/L antisense HSP70 oligomers for 0-100 hr. Cell growth inhibition was analyzed using a trypan blue dye exclusion test. Apoptotic cells were detected and confirmed by flow cytometric analysis and DNA fragmentation analysis. The protein expression of HSP70 and bcl-2 affected by antisense HSP70 oligomers were determined using Western blot. RESULTS: Antisense HSP70 oligomer induced apoptosis and then inhibited proliferation of PC-3m cells in a dose- and time-dependent manner. Ladder-like patterns of DNA fragments were observed in PC 3m cells treated with 10 mmol/L antisense HSP70 oligomer for 48 hr or 8 mmol/L for 72 hr on agarose gel electrophoresis. Antisense HSP70 oligomer pretreatment enhanced the subsequent induction of apoptosis by heat shock in PC-3m cells. In addition, undetectable HSP70 expression was observed at a concentration of 10 mmol/L antisense HSP70 oligomer treatment for 48 hr or 8 mmol/L for 72 hr in Western blot, which was paralleled by decreased expression levels of anti apoptotic protein bcl-2. CONCLUSION: HSP70 antisense oligomer treatment abrogates the expression of HSP70, which may disrupt HSP70-bcl-2-interactions and further down-regulate bcl-2 expression, in turn inducing apoptosis and inhibiting cell growth in PC-3m cells. PMID- 15546022 TI - Relationship between acrosin activity of human spermatozoa and oxidative stress. AB - AIM: To study the association between seminal oxidative stress and human sperm acrosin activity. METHODS: It is a prospective study consisting of 30 infertile men and 12 fertile normozoospermic volunteers. A full history, clinical examination and scrotal ultrasound were done to exclude other related factors such as smoking and varicocele. Presence of white blood cells (WBCs) in semen samples was evaluated by peroxidase staining. Lipid peroxidation in spermatozoa was induced after incubating with ferrous sulphate (4 mmol/L) and sodium ascorbate (20 mmol/L). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS). Acrosin activity was measured using the gelatinolysis technique. The halo diameters around the sperm heads and the percentages of spermatozoa showing halo formation were evaluated. An acrosin activity index was calculated by multiplying the halo diameter by the halo formation rate. RESULTS: A significant difference was observed in acrosin activity parameters and TBARS levels between samples with WBCs (1 multiply 10(6)/mL of ejaculate) and those without. This difference was also noted between the normozoospermic and the oligoasthenoteratozoospermic semen samples. The TBARS production by spermatozoa had a significant negative correlation with the acrosin activity index (r = -0.89, P 0.001). CONCLUSION: The presence of oxidative stress in an individual with leukocytospermia and/or abnormal semen parameters is associated with impaired sperm function as measured by its acrosin activity. PMID- 15546024 TI - Effect of prenatal exposure to diethylstilbestrol on gubernacular development in fetal male mice. AB - AIM: To study the effect of prenatal exposure to diethylstilbestrol (DES) and the role of actin and proliferating cell nuclear antigen (PCNA) on testicular gubernaculum development in fetal male Kunming mice. METHODS: Pregnant mice were randomly assigned to 6 groups and injected with DES subcutaneously from gestational day 9 (E9) to day 17 (E17) at doses of 0, 25, 50, 100, 200 microg.kg 1.d-1 in 0.2 mL dimethyl sulfoxide (DMSO). On E17 they were sacrificed and fetuses quickly removed for fixation. Male fetuses were sliced on serial coronal plane. Histological changes were observed under the light microscope (LM) and ultrastructural changes with the scanning and transmission electron microscopes (SEM and TEM). The expression intensity of actin and PCNA in the gubernacula was quantitated by immunohistochemistry. RESULTS: The mortality of the fetuses was higher in the DES-treated groups than that in the DMSO and saline controls (P<0.05). Under LM the gubernacula were seen to be poorly developed with smaller bulbs. On SEM the bulbs lose the clear demarcation between the mesenchymal inner core and the muscular outer layer and looked like a small cone instead of the normal cylindrical appearance. On TEM there were some smaller disordered myofibrils and sparse cytoplasmic organelles in the gubernacular muscular cells of the treated groups. The expression intensity of actin and PCNA in the gubernacula was significantly weaker in the treated groups than that in the DMSO and saline controls (P<0.05). CONCLUSION: DES induces underdevelopment of the gubernacula in a dose-dependent manner in fetal male mice and down regulates the actin and PCNA expression. PMID- 15546025 TI - Radio-protective effect of vitamin E on spermatogenesis in mice exposed to gamma irradiation: a flow cytometric study. AB - AIM: To investigate the effect of vitamin E on the radioprotection of spermatogenesis and chromatin condensation of spermatozoa during passage through the epididymis in mice exposed to irradiation. METHODS: Adult outbred male ICR mice were orally administered natural vitamin E (VE, D-alpha-tocopheryl acetate) at 400 IU/kg for 7 days before exposure to 1 Gy of gamma-irradiation. The animals were sacrificed at day 1, 7, 14, 21, 28, 35 and 70 post-irradiation (IR) and the percentage of testicular germ cells and epididymal sperm chromatin condensation was analyzed using flow cytometry. RESULTS: Serum D-alpha-tocopheryl acetate levels were 47.4+/-3.2 microg/dL in the treated group, yet it could not be detected in the control group. The testicular weight of irradiated mice pretreated with VE+IR was significantly (P<0.05) higher than that of those without VE treatment (IR) at day 14 and 21 post-irradiation. The percentage of primary spermatocytes (4C) in the VE+IR group was comparable to the controls but significantly (P<0.05) higher than those in the IR group from day 7 to 35 post irradiation. The percentage of round spermatids (1C) in the VE+IR group was also significantly (P<0.05) higher than those in the IR group at day 28 post irradiation. The primary spermatocytes : spermatogonia ratio in the IR group was significantly (P<0.05) declined at day 7 to 35 post-irradiation when compared to the VE+IR and control groups. The round spermatid : spermatogonia ratio in the VE+IR group was significantly (P<0.05) higher than that of the IR group at day 14 and 28 post-irradiation. The chromatin condensation of epididymal spermatozoa measured by propidium iodide uptake was not affected by 1 Gy of gamma irradiation. CONCLUSION: The administration of VE prior to irradiation protects spermatogenic cells from radiation. PMID- 15546026 TI - Joint action of phoxim and fenvalerate on reproduction in male rats. AB - AIM: To evaluate the joint action of phoxim and fenvalerate on the reproductive function in male Sprague-Dawley rats. METHODS: The 2 x 2 factorial analysis experiment was used in the study. The pesticides were orally given at daily doses of phoxim (Pho) 8.2 mg/kg, fenvalerate (Fen) 3.3 mg/kg and Pho 8.2+Fen 3.3 mg/kg (Pho:Fen = 5:2), 5 days a week for 60 days. Sperm motility was measured with computer-assisted sperm motility analysis (CASA) and daily sperm production estimated. Immunoenzymatic method and electron microscopy were used to evaluate the serum testosterone and the testicular morphology, respectively. RESULTS: There were significant decreases in sperm motility parameters in the treated animals, including straight line velocity (VSL), beat cross frequency (BCF), linearity (LIN) and straightness (STR). After treated with Fen, significant decreases in VSL, LIN and STR were demonstrated. Significant decreases of daily sperm production were seen in animals treated with Pho and Pho+Fen in comparison with the controls. Serum testosterone levels were not significantly changed in the treated groups. Factorial ANOVA showed that no significant interactions were noted between Pho and Fen in sperm motility, sperm production and serum testosterone. Both the single and mixed pesticides caused various degrees of testicular lesions, involving vacuolation of endoplasmic reticulum and necrosis of Sertoli cells. CONCLUSION: The pesticides may cause sperm motility changes and testicular lesions in male rats. The action of Pho and Fen may be additive. PMID- 15546027 TI - Prevalence and risk factors of erectile dysfunction in three cities of China: a community-based study. AB - AIM: To determine the age-adjusted prevalence of erectile dysfunction (ED) in 3 big cities of China and to explore its potential sociodemographic, medical and lifestyle correlates. METHODS: A cross-sectional, population-based survey was conducted in three cities of China. Structured questionnaires were administered to 2 226 men, aged 20 - 86 years, by trained interviewers. RESULTS: The age adjusted prevalence of ED was 28.34 % (mild 15.99 %, moderate 7.14 %, severe 5.21 %). In the men above 40, the prevalence was 40.2 %. Age was positively correlated with ED (P<0.01). Education was negatively correlated with ED (P<0.01). Spouse companionship, living condition were positively correlated with ED (P<0.01). Histories of cardiovascular disease, diabetes, and hyperlipidemia were positively correlated with ED (P<0.01). Cigarette smoking was not correlated with ED (P>0.05), while the cigarette consumption and duration were positively correlated with ED (P<0.01). Alcohol drinking is negatively correlated with ED (P<0.01). The duration of drinking was positively correlated with ED (P<0.01). Weekly alcohol consumption was not correlated with ED (P>0.05). CONCLUSION: The prevalence of ED increased with age. Cardiovascular disease, diabetes and hyperlipidemia were positively correlated with the increased prevalence. Sociodemographic and lifestyle factors, such as education, spouse companionship, living condition, cigarette and alcohol consumption or duration also have association with the prevalence of ED. PMID- 15546028 TI - Plasma transforming growth factor-beta1 levels in patients with erectile dysfunction. AB - AIM: To evaluate the plasma TGF-beta1 level in erectile dysfunction (ED) patients of various causes. METHODS: Sixty-two patients with ED and 26 potent men were subjected to the study. Based on multidisciplinary work-ups, including medical history, physical examinations, blood tests with lipid profile and hormones, penile duplex Doppler ultrasonogram and neurophysiological tests, causes for ED were classified as psychogenic (n=15), neurogenic (n=16) and vasculogenic (n=31). The plasma TGF-beta1 level was measured by the ELISA method. RESULTS: The plasma TGF-beta1 level was significantly increased in the ED group (6.7+/-4.9 ng/mL), compared to the control (4.0 +/-2.1 ng/mL) (P<0.01). In the ED groups, there was a significant increase in the vasculogenic group (9.0 +/-5.5 ng/mL), compared to the psychogenic (3.8 +/-1.8 ng/mL) and neurogenic groups (4.8+/-3.2 ng/mL) (P<0.01). Of the vascular risk factors, both the smoking (7.5 +/-4.7 ng/mL) and dyslipidemia groups (7.4+/-4.4 ng/mL) showed significantly increased plasma TGF beta1 levels, compared to the non-smokers (5.5+/-2.8 ng/mL), and those without dyslipidemia (4.8+/-2.8 ng/mL) (P<0.05). CONCLUSION: Vascular risk factors are associated with an elevated plasma TGF-beta1 level, which may contribute to cavernous fibrosis and ED. PMID- 15546029 TI - Relationship between metabolic syndrome and erectile dysfunction. AB - AIM: To determine the relationship between metabolic syndrome (MS) and erectile dysfunction (ED) and to see which risk factors correlated the best with ED. METHODS: Seventy-nine cardiology clinic outpatients with coronary artery disease (CAD) and lipid metabolism disorder were recruited. They were categorized as having MS, hypertension (blood pressure greater than 130/85 mmHg) and dyslipidemia. ED was classified based on International Index of Erectile Function scores. Patients were grouped into quartiles based on body mass index (BMI). Chi square, Pearson's correlation and regression tests were used for statistical analysis. RESULTS: The mean age of the patients was 56.6 years. ED was diagnosed in 59 (74.7 %) of the 79 patients. In the 38 patients with MS, all had ED. ED was not significantly correlated with cholesterol levels (P>0.05), but was found often in patients who had both hypercholesterolemia and HT (P<0.01). Nineteen (76 %) of the 25 patients who had dyslipidemia had ED. However, ED was not significantly correlated with dyslipidemia (P >0.05). Twenty-two of the 23 patients who had BMI greater than 30 had ED, which was significantly more prevalent than that in those who had normal BMI (P<0.01). ED was seen in 38 of 53 smoker patients. Although ED was more prevalent in cigarette smokers, it was not significantly different from non-smokers (P>0.5). CONCLUSION: ED is present in a high percentage of patients with MS. Among multiple risk factors for ED, MS correlates the most highly. The next most important risk group is the patients with hypertension +hypercholestrolemia and obesity (BMI 30). PMID- 15546030 TI - Cycloheximide prevents production of arresting, a fraction of 30-50 kDa obtained from seminiferous tubule conditioned medium. AB - AIM: To evaluate the effect of a protein synthesis inhibitor cycloheximide on arresting activity in spermatogenesis and sperm count in male rats. METHODS: The study used seminiferous tubule (ST) segments from adult rats cultured in vitro with or without cycloheximide to condition culture media, which have been concentrated, size fractioned (30-50 kDa) and administered 7 days to adult rats by intraperitoneal injections. The effects on testicular and epididymal weights, spermatogenesis and epididymal sperm count were determined. RESULTS: The fraction (30-50 kDa), named arresting, obtained from the culture without cycloheximide decreased testicular and epididymal weights (P<0.01) and reduced the epididymal sperm count significantly. Study of the spermatogenic cycle by transillumination showed spermatogenic arrest at stage VII in rats treated with arresting compared to that observed in controls. The length of stage VII in the group receiving the seminiferous tubules culture media with cycloheximide (30-50 KDa CHX-STCM fraction) was similar to control. CONCLUSION: The difference in the effect may be the result of the presence or absence of arresting, a protein secreted by the tubules. PMID- 15546031 TI - Ultrastructural changes of penile tunica albuginea in diabetic rats. AB - AIM: To clarify the ultrastructural changes of penile tunica albuginea (TA) in streptozotocin (STZ)-induced diabetic rats. METHODS: Intraperitoneal injection of STZ was used to induce diabetes mellitus (DM) in 12 Sprague Dawley rats. Ten rats (age and weight-matched) were used as control. Blood samples from the tail snips of the rats were used for the determination of serum glucose levels with SureStep Plus Blood Meter. At week 4 and 10 after the injection, half of the rats in each group were sacrificed and penile samples were obtained from the middle third of the penile shaft for the examination of TA under scanning electron microscopy. RESULTS: In the diabetic group, the serum glucose levels were higher (P<0.01 at both time points) and the TA were thinner (P<0.05) than those of the controls. In the control group, the fibers of TA were rich and arranged regularly and undulated, while in the diabetic group, the fibers were diminished, lost the undulations and were arranged irregularly. CONCLUSION: In rats, DM appeared to impair the penile TA ultrastructures and this impairment could contribute to diabetic erectile dysfunction in part by impairing the veno-occlusive function. PMID- 15546032 TI - Clinical and pathological correlation of the microdeletion of Y chromosome for the 30 patients with azoospermia and severe oligoasthenospermia. AB - AIM: To review the accumulated 30 patients with different area of Y chromosome microdeletions, focusing on their correlation with the clinical and pathological findings. METHODS: A total of 334 consecutive infertile men with azoospermia (218 patients) and severe oligoasthenospermia (116 patients) were screened. Complete physical and endocrinological examinations, general chromosome study and multiplex polymerase chain reaction assay to evaluate the Y chromosome microdeletion were performed. Ten patients received testicular biopsy. Then the clinical and pathological findings were analyzed with reference to the areas of Y chromosome microdeletion. RESULTS: There is a decline of the percentage of sperm appearing in semen in the group that the gene deletion region from AZFc to AZFb. The clinical evidence of the impairment (decreased testicular size and elevated serum FSH) is also relevantly aggravated in this group. However, the pathology of testicular biopsy specimen was poorly correlated with the different deletion areas of the Y chromosome, which may be due to the limited number of specimens. CONCLUSION: The clinical correlation of spermatogenic impairment to the different AZF deletion regions may provide the information for the infertile couples in pre treatment counseling. PMID- 15546033 TI - Severe burn of penis caused by excessive short-wave diathermy. PMID- 15546034 TI - Acute scrotum due to arterial bleeding mimicking non-seminomatous germ cell tumor. AB - Men with testicular tumors usually present with painless increase in testis size incidentally noticed by the patient. We report a case of a young patient presenting as an emergency with acute onset of massive right-sided testicular pain without previous injury. After physical examination testicular torsion could not be excluded. Ultrasound examination of the testis was suspicious for testicular tumor. Surgical exploration of the right testis by inguinal approach was performed revealing subcapsular arterial bleeding due to a small nonseminomatous germ cell tumor non-palpable on clinical examination. PMID- 15546035 TI - Anemia in patients on combined androgen block therapy for prostate cancer. AB - AIM: To study the effect of combined androgen block therapy on hemoglobin and hematocrit values in patients with prostate cancer. METHODS: One hundred and thirty-six patients with adenocarcinoma of prostate were treated with combined androgen block (orchiectomy and flutamide 250 mg, tid). Complete blood counts were determined before and after 1, 2, 3, 6, 9 and 12 months of therapy. RESULTS: The hemoglobin and hematocrit levels declined significantly in all patients and at all the time points after treatment (P<0.05). CONCLUSION: Prostate cancer patients treated with combined androgen block would develop obvious anemia. Recombinant human erythropoietin can be used to treat patients with severe anemia. PMID- 15546036 TI - Prostant in the treatment of chronic prostatitis: a meta-analysis. PMID- 15546037 TI - The biogeography and phylogeny of unicellular cyanobacterial symbionts in sponges from Australia and the Mediterranean. AB - The distribution, host associations, and phylogenetic relationships of the unicellular cyanobacterial symbionts of selected marine sponges were investigated with direct 16s rDNA sequencing. The results indicate that the symbionts of the marine sponges Aplysina aerophoba, Ircinia variabilis, and Petrosia ficiformis from the Mediterranean, four Chondrilla species from Australia and the Mediterranean, and Haliclona sp. from Australia support a diversity of symbionts comprising at least four closely related species of Synechococcus. These include the symbionts presently described as Aphanocapsa feldmannii from P. ficiformis and Chondrilla nucula. A fifth symbiont from Cymbastela marshae in Australia is an undescribed symbiont of sponges, related to Oscillatoria rosea. One symbiont, Candidatus Synechococcus spongiarum, was found in diverse sponge genera in the Mediterranean Sea and the Indian, Pacific, and Southern oceans, whereas others were apparently more restricted in host association and distribution. These results are discussed in terms of the biodiversity and biogeographic distributions of cyanobacterial symbionts. PMID- 15546038 TI - Microbial communities associated with electrodes harvesting electricity from a variety of aquatic sediments. AB - The microbial communities associated with electrodes from underwater fuel cells harvesting electricity from five different aquatic sediments were investigated. Three fuel cells were constructed with marine, salt-marsh, or freshwater sediments incubated in the laboratory. Fuel cells were also deployed in the field in salt marsh sediments in New Jersey and estuarine sediments in Oregon, USA. All of the sediments produced comparable amounts of power. Analysis of 16S rRNA gene sequences after 3-7 months of incubation demonstrated that all of the energy harvesting anodes were highly enriched in microorganisms in the delta Proteobacteria when compared with control electrodes not connected to a cathode. Geobacteraceae accounted for the majority of delta-Proteobacterial sequences or all of the energy-harvesting anodes, except the one deployed at the Oregon estuarine site. Quantitative PCR analysis of 16S rRNA genes and culturing studies indicated that Geobacteraceae were 100-fold more abundant on the marine-deployed anodes versus controls. Sequences most similar to microorganisms in the family Desulfobulbaceae predominated on the anode deployed in the estuarine sediments, and a significant proportion of the sequences recovered from the freshwater anodes were closely related to the Fe(III)-reducing isolate, Geothrix fermentans. There was also a specific enrichment of microorganisms on energy harvesting cathodes, but the enriched populations varied with the sediment/water source. Thus, future studies designed to help optimize the harvesting of electricity from aquatic sediments or waste organic matter should focus on the electrode interactions of these microorganisms which are most competitive in colonizing anodes and cathodes. PMID- 15546039 TI - Bacterial density and community structure associated with aggregate size fractions of soil-feeding termite mounds. AB - The building and foraging activities of termites are known to modify soil characteristics such as the heterogeneity. In tropical savannas the impact of the activity of soil-feeding termites ( Cubitermes niokoloensis) has been shown to affect the properties of the soil at the aggregate level by creating new soil microenvironments (aggregate size fractions) [13]. These changes were investigated in greater depth by looking at the microbial density (AODC) and the genetic structure (automated rRNA intergenic spacer analysis: ARISA) of the communities in the different aggregate size fractions (i.e., coarse sand, fine sand, coarse silt, fine silt, and dispersible clays) separated from compartments (internal and external wall) of three Cubitermes niokoloensis mounds. The bacterial density of the mounds was significantly higher (1.5 to 3 times) than that of the surrounding soil. Within the aggregate size fractions, the termite building activity resulted in a significant increase in bacterial density within the coarser fractions (>20 mum). Multivariate analysis of the ARISA profiles revealed that the bacterial genetic structures of unfractionated soil and soil aggregate size fractions of the three mounds was noticeably different from the savanna soil used as a reference. Moreover, the microbial community associated with the different microenvironments in the three termite mounds revealed three distinct clusters formed by the aggregate size fractions of each mound. Except for the 2-20 mum fraction, these results suggest that the mound microbial genetic structure is more dependent upon microbial pool affiliation (the termite mound) than on the soil location (aggregate size fraction). The causes of the specificity of the microbial community structure of termite mound aggregate size fractions are discussed. PMID- 15546040 TI - Biological soil crusts of sand dunes in Cape Cod National Seashore, Massachusetts, USA. AB - Biological soil crusts cover hundreds of hectares of sand dunes at the northern tip of Cape Cod National Seashore (Massachusetts, USA). Although the presence of crusts in this habitat has long been recognized, neither the organisms nor their ecological roles have been described. In this study, we report on the microbial community composition of crusts from this region and describe several of their physical and chemical attributes that bear on their environmental role. Microscopic and molecular analyses revealed that eukaryotic green algae belonging to the genera Klebsormidium or Geminella formed the bulk of the material sampled. Phylogenetic reconstruction of partial 16S rDNA sequences obtained from denaturing gradient gel electrophoresis (DGGE) fingerprints also revealed the presence of bacterial populations related to the subclass of the Proteobacteria, the newly described phylum Geothrix/ Holophaga/ Acidobacterium, the Cytophaga/ Flavobacterium/ Bacteroides group, and spirochetes. The presence of these crusts had significant effects on the hydric properties and nutrient status of the natural substrate. Although biological soil crusts are known to occur in dune environments around the world, this study enhances our knowledge of their geographic distribution and suggests a potential ecological role for crust communities in this landscape. PMID- 15546041 TI - Genetic diversity of bacterial communities of serpentine soil and of rhizosphere of the nickel-hyperaccumulator plant Alyssum bertolonii. AB - Serpentine soils are characterized by high levels of heavy metals (Ni, Co, Cr), and low levels of important plant nutrients (P, Ca, N). Because of these inhospitable edaphic conditions, serpentine soils are typically home to a very specialized flora including endemic species as the nickel hyperaccumulator Alyssum bertolonii. Although much is known about the serpentine flora, few researches have investigated the bacterial communities of serpentine areas. In the present study bacterial communities were sampled at various distances from A. bertolonii roots in three different serpentine areas and their genetic diversity was assessed by terminal restriction fragment length polymorphism (T-RFLP) analysis. The obtained results indicated the occurrence of a high genetic diversity and heterogeneity of the bacterial communities present in the different serpentine areas. Moreover, TRFs (terminal restriction fragments) common to all the investigated A. bertolonii rhizosphere samples were found. A new cloning strategy was applied to 27 TRFs that were sequenced and taxonomically interpreted as mainly belonging to Gram-positive and alpha-Proteobacteria representatives. In particular, cloned TRFs which discriminated between rhizosphere and soil samples were mainly interpreted as belonging to Proteobacteria representatives. PMID- 15546042 TI - Microbial community structure and oxidative enzyme activity in nitrogen-amended north temperate forest soils. AB - Large regions of temperate forest are subject to elevated atmospheric nitrogen (N) deposition which can affect soil organic matter dynamics by altering mass loss rates, soil respiration, and dissolved organic matter production. At present there is no general model that links these responses to changes in the organization and operation of microbial decomposer communities. Toward that end, we studied the response of litter and soil microbial communities to high levels of N amendment (30 and 80 kg ha(-1) yr(-1)) in three types of northern temperate forest: sugar maple/basswood (SMBW), sugar maple/red oak (SMRO), and white oak/black oak (WOBO). We measured the activity of extracellular enzymes (EEA) involved directly in the oxidation of lignin and humus (phenol oxidase, peroxidase), and indirectly, through the production of hydrogen peroxide (glucose oxidase, glyoxal oxidase). Community composition was analyzed by extracting and quantifying phospholipid fatty acids (PLFA) from soils. Litter EEA responses at SMBW sites diverged from those at oak-bearing sites (SMRO, BOWO), but the changes were not statistically significant. For soil, EEA responses were consistent across forests types: phenol oxidase and peroxidase activities declined as a function of N dose (33-73% and 5-41%, respectively, depending on forest type); glucose oxidase and glyoxal oxidase activities increased (200-400% and 150-300%, respectively, depending on forest type). Principal component analysis (PCA) ordinated forest types and treatment responses along two axes; factor 1 (44% of variance) was associated with phenol oxidase and peroxidase activities, factor 2 (31%) with glucose oxidase. Microbial biomass did not respond to N treatment, but nine of the 23 PLFA that formed >1 mol% of total biomass showed statistically significant treatment responses. PCA ordinated forest types and treatment responses along three axes (36%, 26%, 12% of variance). EEA factors 1 and 2 correlated negatively with PLFA factor 1 ( r = -0.20 and -0.35, respectively, n = 108) and positively with PLFA factor 3 ( r = +0.36 and +0.20, respectively, n = 108). In general, EEA responses were more strongly tied to changes in bacterial PLFA than to changes in fungal PLFA. Collectively, our data suggests that N inhibition of oxidative activity involves more than the repression of ligninase expression by white-rot basidiomycetes. PMID- 15546043 TI - Degradation of 4-chlorophenol at low temperature and during extreme temperature fluctuations by Arthrobacter chlorophenolicus A6. AB - Low average temperatures and temperature fluctuations in temperate soils challenge the efficacy of microbial strains used for clean up of pollutants. In this study, we investigated the cold tolerance of Arthrobacter chlorophenolicus A6, a microorganism previously shown to degrade high concentrations of 4 chlorophenol at 28 degrees C. Luciferase activity from a luc-tagged derivative of the strain (A6L) was used to monitor the metabolic status of the population during 4-chlorophenol degradation. The A6L strain could degrade 200-300 mug mL( 1) 4-chlorophenol in pure cultures incubated at 5 degrees C, although rates of degradation, growth and the metabolic status of the cells were lower at 5 degrees C compared to 28 degrees C. When subjected to temperature fluctuations between 5 and 28 degrees C, A6L continued to degrade 4-chlorophenol and remained active. In soil microcosm experiments, the degradation rates were significantly faster the first week at 28 degrees C, compared to 5 degrees C. However, this difference was no longer seen after 7 days, and equally low 4-chlorophenol concentrations were reached after 17 days at both temperatures. During 4-chlorophenol degradation in soil, CFU and luciferase activity values remained constant at both 5 and 28 degrees C. However, once most of the 4-chlorophenol was degraded, both values decreased by 1-1.5 logarithmic values at 28 degrees C, whereas they remained constant at 5 degrees C, indicating a high survival of the cells at low temperatures. Because of the ability of A. chlorophenolicus A6 to degrade high concentrations of 4-chlorophenol at 5 degrees C, together with its tolerance to temperature fluctuations and stress conditions found in soil, this strain is a promising candidate for bioaugmentation of chlorophenol-contaminated soil in temperate climates. PMID- 15546044 TI - Degradation of 2,4-dichlorophenoxyacetic acid (2,4-d) by a hypersaline microbial mat and related functional changes in the mat community. AB - Microbial mats possibly possess degradation capacities for haloorganic pollutants because of their wide range of different functional groups of microorganisms combined with extreme diurnal changes in pH, oxygen, and sulfide gradients. In this study, 20 mg/l of the chlorinated herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was applied to a pristine hypersaline cyanobacterial mat from Guerrero Negro, Mexico, under a light regime of 12 h dark/12 h light (600 mumol photons/m(2)s). The loss of 2,4-D was followed by chemical GC analysis; functional changes within the mat were determined with microelectrodes for oxygen, photosynthesis, pH, and sulfide. The depletion of 2,4-D due to photooxidation or sorption processes was checked in control experiments. Within 13 days, the light/dark incubated mats degraded 97% of the herbicide, while in permanent darkness only 35% were degraded. Adsorption of 2,4-D to the mat material, agar, or glass walls was negligible (4.6%), whereas 21% of the herbicide was degraded photochemically. The 2,4-D removal rate in the light/dark incubations was comparable to values reported for soils. The phototrophic community of the mat was permanently inhibited by the 2,4-D addition by 17% on average. The sulfate reduction in the entire mat and the respiration in the photic zone were inhibited more strongly but returned to original levels. Since at the end of the experiment the photosynthetic and respiratory activity of the mats were almost as high as in the beginning and 2,4-D almost completely disappeared, we conclude that the examined mats represent a robust and effective system for the degradation of the herbicide where probably the aerobic heterotrophic population is a major player in the degradation process. PMID- 15546045 TI - Fine spatial structure of Atlantic hake (Merluccius merluccius) stocks revealed by variation at microsatellite loci. AB - Genetic variation at 5 microsatellite loci was analyzed for European hake Merluccius merluccius sampled from 9 different regions in the Atlantic Ocean and the Mediterranean Sea. Significant genetic differentiation was found between samples, suggesting a fine subdivision of Atlantic and Mediterranean hake stocks. These results are discussed in the context of the decline of demersal fish species, probably due to overfishing. PMID- 15546046 TI - Measurement of EROD activity: caution on spectral properties of standards used. AB - The activity of the enzyme 7-ethoxy-resorufin- O-deethylase (EROD) has been extensively used in biomonitoring studies for more than a decade. Although the analytical procedure is simple, it is often poorly characterized. In this study spectral properties of particular standard compounds used to measure EROD activity (ethoxyresorufin and resorufin, standards from Molecular Probes) were tested in order to optimize excitation and emission wavelengths to be used in the fluorimetric assay of EROD activity. The optimal excitation wavelength for the detection of resorufin was 560 nm. At this wavelength the excitation represents only 37% of its maximum level for ethoxyresorufin, while it represents 86% for resorufin. This allows discrimination between the fluorescence emitted by both standards, favoring the formed resorufin. Our results demonstrate that any analytical work using spectrofluorometry to measure EROD activity should be preceded by precise determination of the spectral characteristics of each set of standards used. PMID- 15546047 TI - Preliminary study on linkage mapping based on microsatellite DNA and AFLP markers using homozygous clonal fish in ayu (Plecoglossus altivelis). AB - A mapping referential family (F(1)) of ayu was produced by crossing a normal diploid male with a homozygous clonal female. A genetic linkage map was constructed using 191 amplified fragment length polymorphism (AFLP) and 4 microsatellite DNA markers. A total of 178 loci were mapped in 36 linkage groups comprising 1659.6 cM, which includes approximately 77.3% to 81.8% of the total genome. As the markers were randomly distributed over the genome, they showed high efficiency for the construction of a wide linkage map. PMID- 15546048 TI - Isolation and characterization of a fucoidan-degrading marine bacterial strain and its fucoidanase. AB - A marine bacterial strain that degraded fucoidan from Kjellmaniella crassifolia (class Phaeophyceae, order Laminariales, family Laminariaceae) was isolated in our laboratory. The strain was gram-negative, ubiquinone 8 was the predominant respiratory quinone, and the GC-content of its genomic DNA was 36%. The cells of the strain were rod-shaped (2.0 microm long x 1.0 microm wide), and each cell was motile by means of one polar flagellum. Phylogenetic analysis of its 16S ribosomal DNA sequence indicated that it was a member of the family Alteromonadaceae. It produced a type of extracellular fucoidanase, an endosulfated fucan-digesting enzyme. The enzyme was purified with 3500-fold purity at 12.0% yield. Optimum conditions for the enzyme reaction were approximately pH 6.5 to 8.0 and temperature 30 degrees to 35 degrees C. The enzyme was activated by calcium ions, and maximum activity was observed in the presence of greater than 30 mM calcium ion. PMID- 15546049 TI - Isolation of new Symbiodinium strains from tridacnid giant clam (Tridacna crocea) and sea slug (Pteraeolidia ianthina) using culture medium containing giant clam tissue homogenate. AB - Recent molecular biological studies have revealed that some photosymbiotic invertebrates dwelling in coral reefs host several genetically different dinoflagellates, Symbiodinium species, as symbionts. However, little is known about the difference in physiologic characteristics among these symbionts living in a single host, because some Symbiodinium strains are difficult to culture in vitro. To isolate some of these Symbiodinium strains, we have developed an agar culture medium plate containing antibiotics and a giant clam tissue homogenate. Using-this medium we isolated two new Symbiodinium strains from two molluscan hosts, Tridacna crocea and Pteraeolidia ianthina, each of which hosted two different Symbiodinium strains belonging to Symbiodinium C and D, respectively. The tissue homogenate was essential for the growth of Symbiodinium D. Although it was not essential for the growth of Symbiodinium C, it did stimulate the initial growth. For the isolation of some Symbiodinium strains, isolation medium containing host homogenate is effective. PMID- 15546050 TI - Suppression subtractive hybridization cDNA libraries to identify differentially expressed genes from contrasting fish habitats. AB - Suppression subtractive hybridization complementary DNA libraries identified differentially expressed genes in liver tissue of winter flounder collected from the highly impacted Raritan-Hudson estuary versus those from less industrialized estuaries farther south in New Jersey. Distinct transcript profiles emerged in the fish from these different habitats. A total of 251 clones from the forward (upregulated with anthropogenic impact) and reverse (downregulated with anthropogenic impact) subtracted libraries were sequenced. In the upregulated library immune response transcripts, including complement C-3, C-7, factor H, factor Bf/C2, differentially regulated trout protein 1, and the antimicrobial hepcidin, indicated the pollution-impacted fish were under a high viral or bacterial load. Transcripts for cytochrome P450 1A, P450 3A, and glutathione S transferase, important components of phase I and II metabolism of xenobiotics, were found in the upregulated-with-pollution library. Vitellogenins I and II and egg envelope protein (zp) appeared to be downregulated. A homologue of the tumor suppressor p33(ING1) (down) and hepatocyte growth factor-like protein (up) may indicate liver damage or hepatocellular carcinoma or hepatoma. These expression patterns, confirmed by quantitative polymerase chain reaction, indicate that transcript analysis is a useful method for assessing the health of local habitats and the organisms therein. PMID- 15546051 TI - [Can the planning process for the mental health system in Germany follow scientific principles?]. PMID- 15546052 TI - [The impact of social welfare policies on the deinstitutionalisation of the mentally ill in Germany]. AB - OBJECTIVE: In Germany the competent institution for supported housing for mentally ill and handicapped persons is traditionally the regional welfare authority, while welfare authorities on state (Lander) level hold the competencies for accommodation in hostels and homes: Assessment of the impact of policies that try to overcome this separation of competencies. METHOD: 1. Analysis of the practise of distributing the competencies for accommodation for mental ill and handicapped persons in the 16 German Lander. 2. Assessment of the supply with accommodation for mental ill and handicapped persons in the Lander. 3. Investigation of a possible relationship between distribution of competencies for accommodation and number of accommodation in supported housing and hostels/homes. RESULTS: Data about available accommodation in supported housing and hostels are inconsistently limited and available. As far as this data can be interpreted, Lander that have the administrative and particularly financial competency for accommodation uniquely organised, generally offer more supported housing. At the same time they provide relatively less accommodation in hostels and homes. Despite having the competencies for accommodation uniquely organised, some Lander continue to provide accommodation for mentally ill persons predominantly in hostels and homes. CONCLUSION: A comprehensive and between the Lander comparable system of documentation of accommodation for mentally ill and handicapped persons is highly needed. Unique organisation of competencies for accommodation of mental ill and handicapped persons is a necessary - however not commensurate precondition for the increase of the supply in supported housing. Beyond that, other factors seem to be influential, such as political will, attitudes towards the mentally ill, interests of hostel providers, pre-existing hostel infrastructure. In respect of these factors more research is needed. PMID- 15546053 TI - [Motor skills and psychiatric disturbances in children]. AB - OBJECTIVE: Within the multiprofessional diagnostic in the psychiatric treatment for children an age-group of in-patients was examined using the coordination test for children (KTK) with regard to physical ability. METHODS: The ascertained results and classifications of the motor skills were related to ICD 10 diagnosis on level 1 (clinical psychiatric syndrom) and level 2 (circumscribed disturbance of the development) as well as to issues of the anamnese- and report documentation in order to ascertain correlations between child psychiatry disorder and motor development. RESULTS: More than 40 % of the children examined showed motor deficiency. Moreover, it was evident that girls attained significantly lower KTK-results than boys. The statistical results did not show any distinct effects to "diagnosis" and "age". CONCLUSIONS: The high quota of obviously impaired and disturbed children reflects the general decline of motor skills and does not seem to be attributed to the demand for psychiatric treatment for children and youth. PMID- 15546054 TI - [Problematic consumption of addictive drugs in psychiatric rehabilitation]. AB - OBJECTIVE: Is an inferior outcome of psychiatric rehabilitation to be expected with patients who consume alcohol or addictive drugs in a problematic manner during rehabilitation treatment? METHOD: Files and final reports from patients admitted to psychiatric rehabilitation during three successive years were analyzed. Data of patients with or without problematic use of psychotropic substances were compared. RESULTS: Patients with a problematic pattern of alcohol or drug consumption have a stronger propensity to quit psychiatric rehabilitation when a) using hard drugs or when b) bearing a diagnosis beyond the schizophrenia spectrum. The outcome of rehabilitation efforts of the subsample with a problematic use of drugs shows an unfavorable tendency concerning housing, but a better result in the vocational area. CONCLUSIONS: The problem has to be considered in a differenciated manner, which means that, in general, clients with a problematic use of alcohol or drugs should not automatically be excluded from psychiatric rehabilitation. PMID- 15546055 TI - [Quality of care in internal medicine in German psychiatric hospitals]. AB - OBJECTIVE: Study on selected aspects of structural quality of care in internal medicine in German psychiatric hospitals. METHOD: Investigation by questionnaire completed by medical directors of German psychiatric hospitals (response quote 48.8 %). RESULTS: Descriptive data from 81 hospitals with 25 920 inpatient beds are presented relating to various indicators of structural quality of care in internal medicine (e. g. availability of internal medicine consultations, diagnostic equipment, as ECG, clinical chemistry laboratory, ultrasound). CONCLUSIONS: We found evidence of a satisfactory infrastructure relating to internal medicine in a majority of hospitals. However, there exist also indications of significant deficits pertaining to certain aspects of medical care in the institutions investigated. With regard to the importance of medical comorbidity in clinical psychiatry the availability of adequate resources in internal medicine is a relevant indicator of quality of care in psychiatric hospitals. PMID- 15546056 TI - [Acute psychiatric day hospital treatment: is the effectiveness of this treatment approach still questionable?]. AB - OBJECTIVE: Currently, there is still a severe lack of methodologically sound empirical studies on acute psychiatric day hospital treatment in German-speaking countries that analyse the effectiveness of this increasingly important mode of service provision. METHODS: Within a randomised controlled study design implemented at the Department of Psychiatry and Psychotherapy, Dresden University of Technology, 99 general psychiatric patients received conventional inpatient treatment and 92 patients received acute day hospital treatment. At up to four time-points during the index-treatment episode patients were assessed at different levels of outcome: Psychopathology was rated by researchers using the Brief Psychiatric Rating Scale (24-Item-Version), and patients evaluated their satisfaction with treatment (Patientenbogen zur Behandlungszufriedenheit); at admission and discharge patients also assessed their subjective quality of life (Manchester Assessment of Quality of Life). Mean scale scores of these instruments are used for the intention-to-treat-analysis. Discharge status on these scales as well as mean ratings on these scales within the index-treatment episode serve as measures of effectiveness. For statistically identifying differences between the two settings five linear (co-)variance-analytical models were calculated for each target variable. Four models were adjusted to baseline rating or to the individual period spent in treatment. RESULTS: Initially, both groups did not differ in their relevant socio-demographic and illness-related features. Day hospital treatment (87,7 days) lasted significantly longer than inpatient treatment (67,8 days). Only results from an unadjusted statistical model as well as from a model adjusted to the individual period of index hospitalisation demonstrated superior effectiveness of day hospital treatment on the discharge status of psychopathological symptomatology. However, in all statistical models there were no systematic differences of treatment effectiveness related to satisfaction with treatment and subjective quality of life. CONCLUSION: For the first time in German-speaking countries, this study provides evidence for the effectiveness of acute day hospital treatment as compared to conventional inpatient treatment. If detailed eligibility criteria for patients are used as defined here, approximately 30 % of the general psychiatric patients in need of acute hospital-based treatment may be cared for in this special mode of day hospital service provision. PMID- 15546057 TI - [Social psychiatry without sociology. Analysis of a scientific journal]. AB - AIM: Psychiatry and Sociology have followed separate avenues in Germany since the mid 1970 s. The aim of this work is to investigate their present relationship by means of an analysis of a scientific journal. METHOD: All original works dealing with social psychiatric subjects and appearing in Psychiatrische Praxis in the years 2002 and 2003 were analysed. RESULTS: Sociologists appear to be a minority among the authors. Questions that are explicitly related to sociological theories were an exception, and sociological authors have hardly been quoted. CONCLUSION: Social psychiatric research in Germany is still conducted without Sociology. If social psychiatry is to gain in importance as a scientific discipline, closer cooperation with its mother discipline is imperative. As an example, the application of the rational choice theory in psychiatric research is discussed. PMID- 15546059 TI - Course typologies, treatment principles, and research concepts. AB - In order to explore the mechanisms of illness course and outcome and to improve treatment in schizophrenia and affective disorders, a theoretical framework as provided by the biopsychosocial concept, e. g. the vulnerability-stress-coping model, has been applied. To develop a coherent and valid research strategy, however, respective model components have to be translated into neuroscience concepts and operationalized accordingly. Course characteristics (typologies, markers), course influences (determinants, triggers, modifiers) and course predictors are discussed from a theoretical perspective. Building on this discussion, an integrative concept of illness course and course mechanisms is proposed. Finally, further research recommendations are given. Future research should elaborate on the neurobiological characteristics of course and outcome in methodologically adequate prospective interventional long-term studies. PMID- 15546060 TI - Mathematical-statistical concepts for modelling and prediction of longterm follow up. AB - Newer statistical methods for modeling and prediction of long-term follow-up in schizophrenia are presented. These include the extended Cox model, the Generalized Estimating Equations (GEE) method and the Artificial Neural Networks (ANN) approach. PMID- 15546061 TI - The etiopathogenesis of schizophrenias. AB - There is no generally accepted model explaining the aetiopathogenesis of schizophrenias. In recent years, hypotheses with a focus on single aspects of neurotransmission, single major genetic loci or single brain areas were predominating. Now, these different approaches converge to systemic models including neurotransmission, genetic changes and neurodevelopmental malformation of brain structures. Such systemic models will explain more aspects of schizophrenias than the recent confined hypotheses. PMID- 15546062 TI - Effect of chronic antidepressant treatment on beta-receptor coupled signal transduction cascade. Which effect matters most? AB - BACKGROUND: Beta-receptor down-regulation has been described as a common biochemical effect of chronic treatment with many but not all antidepressant drugs. Beta-receptor activation leads to elevated intracellular levels of cAMP followed by the activation of several protein kinases which in turn activate various transcription factors. One of those, CREP has received increasing interest as an relevant component within the antidepressant drug modulated signal cascade as it represents a down-stream signal not only of the beta-receptor but also of serotonin receptor activation. Chronic treatment with many antidpressant drugs has been shown to alter CREP levels in several brain regions. While beta receptor down-regulation by chronic antidepressant treatment has been a consistent finding, alterations of CREP levels have been observed in both direction. Similarly divergent findings have been reported for BDNF a major gene targeted of CREB, where most but not all findings suggest up-regulation at least at the message level following chronic antidepressant treatment. METHODS: Because of these rather divergent data, we investigated the possible effects of chronic treatment (9 or 19 days) with three different antidepressant drugs (reboxetine, citalopram, imipramine) on the individual parameters of the beta-receptor coupled signal transduction cascade. All animals were also tested for possible antidepressant effects using the forced swimming test. RESULTS: While beta receptor density was down-regulated by reboxetine and imipramine but not citalopram, CREB protein was only mildly elevated after 9 days, and not changed or slightly reduced after 19 days. BDNF protein levels were not or only slightly enhanced, but only for the 9 days treatment. Citalopram was most active. Under the conditions chosen, all three drugs were active in the forced swimming test. CONCLUSION: Taken together, the findings reported make it difficult to identify one single component of the beta-receptor coupled signal transduction cascade as common final target of chronic antidepressant treatment. PMID- 15546063 TI - Concepts on neurological disease evolution. AB - Degenerative neurological diseases evolve progressively with an insidious onset, while relapsing neurological diseases may remit completely or result in progressive deficits. Since the affected systems can be assessed by clinical evaluation and ascribed to disease-specific pathology--today already in vivo using neuroimaging, the mechanisms underlying neurological disease manifestation can also be studied. For the most frequent neurological diseases with a relapsing clinical course, such as multiple sclerosis, brain infarction and epilepsy, it is shown that disease-specific aetiopathogenesis, lesion-specific pathophysiology and unspecific bystanders determine disease manifestation. Nevertheless, prediction of progression or relapse of a neurological disease is poor due to ill defined relations of molecular genetic markers and disease evolution. Recent research on aetiopathogenesis and pathophysiology providing new perspectives for selective therapeutic interventions and relapse prevention is discussed. PMID- 15546064 TI - Course and long-term treatment of schizophrenic psychoses. AB - As there is a lack of clear biological correlates for most psychiatric disorders, follow-up studies regarding course, outcome and prognosis of psychiatric disorders play an important role in psychiatric research, especially in terms of validation of psychiatric diagnoses and other psychiatric concepts, such as the concept of negative symptoms. Long-term studies also have their place in the description and evaluation of first treatment procedures. The first part of this paper will describe some general aspects of the long-term course and outcome of schizophrenic psychoses. The problem of relapses and relapse prevention will then be discussed. Especially data from recent studies will be considered in this overview. PMID- 15546065 TI - Etiopathogenetic mechanisms in long-term course of schizophrenia. AB - Schizophrenia is a severe mental disorder with a relapsing long-term course in 80 % of the sufferers. The underlying neurobiological principles of the long-term course are widely unknown. Therefore an attempt is made to evaluate data from structural imaging and neuropsychology to shed some light on these mechanisms. Interestingly there was a significant correlation between length of illness and volume reduction in the prefrontal grey matter in schizophrenia. There was a lack of such a correlation for the whole brain volume, white matter volume or the grey matter volume outside the prefrontal lobe. Furthermore none of the neuropsychological measures correlated with length of illness. Both findings are supported by data from prospective studies up to five years demonstrating a significant loss of frontal grey matter volume, but no change of cognitive dysfunction in the course of time. The attempt is made to connect prefrontal grey matter loss with post-mortem findings of reduced neuropil but preserved cytoarchitecture leading to recently described candidate genes and their function. PMID- 15546066 TI - Mechanisms of relapse prevention in schizophrenia. AB - INTRODUCTION: A relapse can be expected in 70 % of patients after the first schizophrenic episode; in parallel 70 % of patients show an incomplete remission of the disorder after the first episode. This includes a cognitive decline and persistence of schizophrenic negative symptoms, often associated with social disabilities, social decline and a worsened quality of life. The risk for a relapse after a schizophrenic episode remains increased throughout the patient's lifetime. Moreover, the risk for chronic disorder increases with every relapse of the patient. Therefore relapse prevention is a primary focus in the treatment of schizophrenia. METHODS: Studies point out that atypical antipsychotics have advantages in relapse prevention of schizophrenia compared to the "classical" antipsychotics, although comparative studies are still lacking. RESULTS: The main risk factor for a relapse is the withdrawal of antipsychotic medication. Nonbiological factors such as high-expressed emotions, adverse life events and stress contribute to an increased risk of schizophrenic relapse. Studies of relapse after withdrawal of antipsychotic medication revealed that noradrenergic, serotonergic, and dopaminergic neurotransmission are involved, low serotonergic and high dopaminergic neurotransmission are associated with a relapse, and possibly high noradrenergic neurotransmission plays a role in short-term relapses. Inflammation and the activation of certain cytokines, which have rarely been studied to date, seem to have an even higher impact on schizophrenic relapse. Methodological aspects of these studies are discussed. CONCLUSION: Biological mechanisms of relapse may not become fully elucidated before the mechanisms of the schizophrenic pathophysiology are clarified. PMID- 15546067 TI - Affective disorders: basic principles regarding clinical course, long-term therapeutic and prophylactic strategies. AB - Affective disorders, mainly depressive and bipolar afflictions are life-long recurrent diseases and require constant treatment and prophylaxis. Both acute and maintenance therapeutic strategies have to be applied to the special subgroups of the depressive and bipolar spectrum. Despite the development of new pharmacological agents and psychotherapeutical settings, the long-term prognosis, especially functional outcome, did not change considerably in comparison to past decades. PMID- 15546068 TI - Basic pathophysiological mechanisms in depression: what are they and how might they affect the course of the illness? AB - Basic pathophysiological mechanisms in affective disorders are discussed. Studies carried out suggest that changes in neurogenesis do not underlie the behavioral changes which lead to helplessness. Since the behavioral changes leading to depressive or anxious behaviors are not correlated with changes in neurogenesis it appears unlikely that a decrease in the rate of neurogenesis is the basis for depression. A modified gene expression resulting in both functional and structural brain changes remains the most consistent hypothesis to explain how affective disorders may occur. An alternative candidate, synaptogenesis, appears as a likely candidate and requires further experimental testing. PMID- 15546069 TI - Mechanisms of action in the prevention of recurrent mood disorders. AB - BACKGROUND: Since long-term treatment is a need in many patients with mood disorders, knowledge on mechanisms of action in the prevention of recurrence is of major relevance. METHODS: Follow-up studies, which are best suited to prove the linkage of causal factors and mechanisms with the clinical course of the disorders, are rare. Another approach to search for preventive mechanisms is to address actions of pharmacological agents, which are effective in prophylactic treatment of mood disorders. Studies over the past several years have indicated that intracellular signaling cascades mediate long-term pharmacological effects via modulation of transcription factors and gene expression of neurotrophic and neuroprotective factors which may interfere with recurrence of affective illness. RESULTS: The impact of treatment on stress responsive systems varies with different therapeutic strategies and the relevance of a treatment-related modification of stress reliability for the course of the disease is not clear. Functional and structural actions of mood stabilizers and of antidepressants on mood-relevant anatomical circuits and modulation of chronobiological alterations also appear to be involved in preventive mechanisms in mood disorders. Moreover, cognitive mediators of prophylactic effects have been described. CONCLUSION: Combined clinical and biological studies should assess the relevance of those modes of action for the long-term course of mood disorders. PMID- 15546070 TI - Syphilitic hepatitis in HIV-infected patients: a report of 7 cases and review of the literature. AB - BACKGROUND: A recent resurgence of primary and secondary syphilis has been observed in certain population groups, particularly among persons infected with human immunodeficiency virus (HIV). Liver involvement is an infrequently recognized complication of early syphilis, with no previous reports among HIV infected patients. METHODS: We describe 7 cases of syphilitic hepatitis in HIV positive individuals and review the literature. RESULTS: At our institutions, all patients presented with a rash consistent with secondary syphilis. Each case was characterized by a conspicuous increase in serum alkaline phosphatase level (mean level +/- standard deviation, 905 +/- 523.6 IU/L) and milder elevations in serum transaminase levels. The mean CD4+ absolute T cell count was 317 cells/mm3, and the median rapid plasma reagin (RPR) titer was 1 : 128. There was a significant correlation between higher CD4+ cell counts and the RPR titers (R=0.93; P=.002). Symptomatic resolution and biochemical improvement, particularly a significant decrease in serum alkaline phosphatase levels (P=.02), occurred following antibiotic therapy. CONCLUSIONS: Hepatic dysfunction is not uncommon in HIV infected persons and is attributable to multiple causes. In the appropriate clinical setting, syphilitic hepatitis is an easily diagnosed and reversible etiology of liver dysfunction. The recognition of this entity will prevent unnecessary evaluation of abnormal liver enzyme levels in HIV-positive patients. PMID- 15546071 TI - Two cases of fatal shock after transfusion of platelets contaminated by Staphylococcus aureus: role of superantigenic toxins. AB - We detected Staphylococcus aureus superantigenic toxins in the platelet infusion bags that had been used for 2 patients who subsequently developed transfusion associated Staphylococcus aureus infection. Both patients, who were immunodeficient, developed manifestations of toxic shock syndrome and septic shock, and they died soon after the onset of infection. PMID- 15546072 TI - Successful desensitization to enfuvirtide after a hypersensitivity reaction in an HIV-1-infected man. AB - We report a case of successful, rapid desensitization to enfuvirtide after a hypersensitivity reaction in a man with highly drug-resistant human immunodeficiency virus type 1 infection. The patient was desensitized in a monitored intensive care unit and tolerated the rapid desensitization protocol without any serious adverse effects. This case illustrates the ability to safely desensitize patients with limited treatment options who require enfuvirtide therapy. PMID- 15546073 TI - Micafungin versus fluconazole for prophylaxis against invasive fungal infections during neutropenia in patients undergoing hematopoietic stem cell transplantation. AB - We hypothesized that chemoprophylaxis with the echinocandin micafungin would be an effective agent for antifungal prophylaxis during neutropenia in patients undergoing hematopoietic stem cell transplantation (HSCT). We therefore conducted a randomized, double-blind, multi-institutional, comparative phase III trial, involving 882 adult and pediatric patients, of 50 mg of micafungin (1 mg/kg for patients weighing <50 kg) and 400 mg of fluconazole (8 mg/kg for patients weighing <50 kg) administered once per day. Success was defined as the absence of suspected, proven, or probable invasive fungal infection (IFI) through the end of therapy and as the absence of proven or probable IFI through the end of the 4 week period after treatment. The overall efficacy of micafungin was superior to that of fluconazole as antifungal prophylaxis during the neutropenic phase after HSCT (80.0% in the micafungin arm vs. 73.5% in the fluconazole arm [difference, 6.5%]; 95% confidence interval, 0.9%-12%; P=.03). This randomized trial demonstrates the efficacy of an echinocandin for antifungal prophylaxis in neutropenic patients. PMID- 15546074 TI - An open, randomized, controlled trial of penicillin, doxycycline, and cefotaxime for patients with severe leptospirosis. AB - BACKGROUND: Leptospirosis is an important cause of fever in the rural tropics. Since 1996, there has been a marked increase in the incidence of leptospirosis in northeastern Thailand. Although leptospirosis generally is susceptible to antibiotics, there is no consensus regarding the optimal treatment for severe leptospirosis. METHODS: An open-label, randomized comparison of parenteral cefotaxime, penicillin G sodium (hereafter known as "penicillin G"), and doxycycline for the treatment of suspected severe leptospirosis was conducted. The study involved 540 patients admitted to 4 hospitals in northeastern Thailand. RESULTS: A total of 264 patients (48.9%) had leptospirosis confirmed by serologic testing or culture. The overall mortality rate was 5%. There were no significant differences between the antibiotics with regard to associated mortality, defervescence, or time to resolution of abnormal findings of laboratory tests either among all study participants or among the subgroup of patients with confirmed leptospirosis. A total of 132 patients had rickettsial infection diagnosed, and, for these patients, treatment with doxycycline was superior to treatment with penicillin G. CONCLUSIONS: Doxycycline or cefotaxime is a satisfactory alternative to penicillin G for the treatment of severe leptospirosis. PMID- 15546075 TI - The influence of human N-acetyltransferase genotype on the early bactericidal activity of isoniazid. AB - The elimination of isoniazid is subject to the influence of the N acetyltransferase 2 (NAT2) genotype, and individuals may be homozygotic slow, heterozygotic fast, or homozygotic fast acetylators of isoniazid. The early bactericidal activity (EBA) of an antituberculosis agent can be determined by quantitative culture of Mycobacterium tuberculosis in sputum samples obtained from patients with pulmonary tuberculosis during the first days of treatment. In these studies, the EBA of isoniazid during the first 2 days of treatment was determined for 97 patients with sputum smear-positive pulmonary tuberculosis following isoniazid doses of < or =37.5 mg, 75 mg, 150 mg, 300 mg, and 600 mg. The NAT2 genotype was determined in 70 patients, and the association between EBA and genotype was examined in this subgroup. Similarly, the relationship between EBA and isoniazid serum concentration was evaluated in 87 patients. The mean EBA of isoniazid increased with dose, but it levelled off between doses of 150 mg (mean EBA, 0.572) and 300 mg (mean EBA, 0.553). Significant differences were found in the mean EBA of isoniazid between the homozygous slow acetylator group and the heterozygous fast acetylator group and between the homozygous slow acetylator group and the homozygous fast acetylator group, but not between the heterozygous fast acetylator group and the homozygous fast acetylator group. The EBA appeared to reach a maximum at a 2-h isoniazid concentration of 2-3 microg/mL. These data confirm a significant effect of NAT2 genotype on the EBA of isoniazid over a range of doses. PMID- 15546076 TI - Studies of early bactericidal activity: new insights into isoniazid pharmacokinetics. PMID- 15546077 TI - Longitudinal risk of herpes simplex virus (HSV) type 1, HSV type 2, and cytomegalovirus infections among young adolescent girls. AB - BACKGROUND: Cross-sectional seroprevalence studies indicate that infections with herpes simplex virus (HSV) types 1 (HSV-1) and 2 (HSV-2) and cytomegalovirus (CMV) are common. However, data on the rates of acquisition of these infections are limited. METHODS: A 3-year longitudinal study of HSV-1, HSV-2, and CMV seroprevalence was conducted in a cohort of 174 adolescent girls (age at enrollment, 12-15 years). RESULTS: At study entry, 41% of the girls reported a history of sexual activity, and by the end of the study, 73% reported a history of sexual activity. At enrollment, 71% of all participants were seropositive for CMV, 44% were seropositive for HSV-1, and 7% were seropositive for HSV-2. By the end of the study, 81% of the girls were seropositive for CMV, 49% were seropositive for HSV-1, and 14% were seropositive for HSV-2. Among girls with a history of sexual activity, 15.5% were HSV-2 seropositive at the beginning of the study, and 18.9% were HSV-2 seropositive at the end of the study. The attack rates, based on the number of cases per 100 person-years, were 13.8 for CMV infection and 3.2 for HSV-1 infection (among all girls) and 4.4 for HSV-2 infection (among girls with a history of sexual activity). Participants with preexisting HSV-1 antibodies were associated with a significantly lower attack rate for HSV-2 infection. A generalized estimating equation model indicated that participants with a longer history of sexual activity and those who had more sexually transmitted diseases during the 6-month periods before the study visits were more likely to be HSV-2 seropositive. CONCLUSIONS: This longitudinal study of adolescent girls found high baseline CMV and HSV-1 seroprevalence rates and substantial attack rates for all 3 pathogens. PMID- 15546078 TI - A key role for adolescents in the epidemiology of cytomegalovirus and genital herpes infections. PMID- 15546079 TI - Use of maximal sterile barriers during central venous catheter insertion: clinical and economic outcomes. AB - BACKGROUND: We performed a cost-effectiveness analysis to determine the effect of maximal sterile barriers (MSBs) on reducing central venous catheter (CVC)- related infections. Use of MSBs when placing CVCs may reduce the risk of infections but is more cumbersome, time-consuming, and expensive than other techniques. METHODS: We developed a decision analytic model in which a patient could have a CVC placed with either an MSB or a less stringent technique. We calculated total direct medical costs and the incidences of catheter-related bloodstream infections, catheter colonization, and death. RESULTS: Use of MSBs lowered costs (from 621 dollars to 369 dollars per catheter insertion) and decreased the incidences of catheter-related bloodstream infections (from 5.3% to 2.8%), catheter colonization with local infection (from 5.5% to 2.9%) and death (from 0.8% to 0.4%). MSBs improved patient safety throughout all sensitivity analyses. CONCLUSIONS: Use of MSBs during CVC insertion likely lowers medical costs and decreases the incidences of catheter colonization, catheter-related bloodstream infections, and death. Cost savings were found over a wide range of clinical and economic assumptions, suggesting that MSBs should be routinely used when CVCs are inserted. PMID- 15546080 TI - A high-morbidity outbreak of methicillin-resistant Staphylococcus aureus among players on a college football team, facilitated by cosmetic body shaving and turf burns. AB - BACKGROUND: Athletics-associated methicillin-resistant Staphylococcus aureus (MRSA) infections have become a high-profile national problem with substantial morbidity. METHODS: To investigate an MRSA outbreak involving a college football team, we conducted a retrospective cohort study of all 100 players. A case was defined as MRSA cellulitis or skin abscess diagnosed during the period of 6 August (the start of football camp) through 1 October 2003. RESULTS: We identified 10 case patients (2 of whom were hospitalized). The 6 available wound isolates had indistinguishable pulsed-field gel electrophoresis patterns (MRSA strain USA300) and carried the Panton-Valentine leukocidin toxin gene, as determined by polymerase chain reaction. On univariate analysis, infection was associated (P<.05) with player position (relative risk [RR], 17.5 and 11.7 for cornerbacks and wide receivers, respectively), abrasions from artificial grass (i.e., "turf burns"; RR, 7.2), and body shaving (RR, 6.1). Cornerbacks and wide receivers were a subpopulation with frequent direct person-to-person contact with each other during scrimmage play and drills. Three of 4 players with infection at a covered site (hip or thigh) had shaved the affected area, and these infections were also associated with sharing the whirlpool > or =2 times per week (RR, 12.2; 95% confidence interval, 1.4-109.2). Whirlpool water was disinfected with dilute povidone-iodine only and remained unchanged between uses. CONCLUSIONS: MRSA was likely spread predominantly during practice play, with skin breaks facilitating infection. Measures to minimize skin breaks among athletes should be considered, including prevention of turf burns and education regarding the risks of cosmetic body shaving. MRSA-contaminated pool water may have contributed to infections at covered sites, but small numbers limit the strength of this conclusion. Nevertheless, appropriate whirlpool disinfection methods should be promoted among athletic trainers. PMID- 15546081 TI - Use of stool collection kits delivered to patients can improve confirmation of etiology in foodborne disease outbreaks. AB - BACKGROUND: In 68% of foodborne disease outbreaks, no etiologic pathogen is identified. In two-thirds of outbreaks with no identified etiology, no stool specimens are submitted for testing. METHODS: From April 2001 to March 2003, we pilot-tested use of prepackaged, self-contained stool specimen collection kits in 3 states, delivered to and from patients by courier or mail, to improve rates of specimen collection in the outbreak setting. Specimens were tested for bacterial and viral pathogens at health department laboratories, and results were correlated with epidemiological investigation data. RESULTS: Specimens were returned by > or =1 person in 52 (96%) of 54 outbreaks in which kits were deployed; in total, 263 (76%) of 347 persons who received kits returned specimens. Resolution of symptoms was the most commonly cited reason for nonsubmission of kits. An etiology was confirmed in 37 (71%) of 52 outbreaks with specimens returned; 28 (76%) were attributable to norovirus, and 9 (24%) were attributed to bacterial pathogens. Stool kits were well received and cost an average of approximately 43 dollars per specimen returned. CONCLUSIONS: In two thirds of foodborne disease outbreaks in which delivered stool collection kits were successfully deployed, an etiologic organism was identified. Delivery of kits to and from patients to improve rates of stool collection in outbreaks in which specimens might otherwise not be submitted could substantially reduce the number of outbreaks with an unknown etiology. PMID- 15546082 TI - Emergence of new strains of methicillin-resistant Staphylococcus aureus in a neonatal intensive care unit. AB - BACKGROUND: Genetically distinct strains of methicillin-resistant Staphylococcus aureus (MRSA) of community rather than hospital origin have emerged in many areas of the United States. We determined if MRSA strains causing bacteremia in infants treated from birth in a neonatal intensive care unit (NICU) demonstrated the genetic traits of community-associated MRSA. METHODS: A retrospective cohort study was conducted among NICU infants with bacteremia due to MRSA during 2003 in a large tertiary care center NICU in Houston. MRSA isolates were characterized by antimicrobial susceptibility testing and staphylococcal cassette chromosome mec (SCCmec) typing by polymerase chain reaction. All MRSA cases were reviewed for clinical severity of infection and outcome. RESULTS: During 2003, a total of 8 (47%) of 17 infants with bacteremia due to S. aureus had MRSA infection. Isolates from 6 (75%) of these 8 infants carried the SCCmec genes (class B mec and ccr2) that are characteristic of community MRSA; 4 isolates were type IVa. All 6 isolates were resistant to beta-lactam antibiotics and erythromycin; 1 was also resistant to clindamycin. One isolate was nontypeable, and another carried the SCCmec type II gene (typical of hospital-associated strains) and was susceptible only to vancomycin. Seven (88%) of 8 infants presented in septic shock. Despite initial treatment with vancomycin, 3 (38%) died, and 3 survivors had complications requiring prolonged antimicrobial therapy; these 6 infants had MRSA isolates with genetic characteristics of isolates of community origin. CONCLUSIONS: Community-associated MRSA strains have emerged as a significant cause of sepsis in neonates hospitalized in NICU since birth and have caused disseminated infection with substantial morbidity and mortality. PMID- 15546083 TI - Utility of Aspergillus antigen detection in specimens other than serum specimens. AB - The detection of circulating galactomannan in serum is an important tool for the early diagnosis of invasive aspergillosis. A commercial enzyme-linked immunosorbent assay (Platelia Aspergillus; BioRad) was shown to be both highly sensitive and specific for detection of galactomannan in serum samples. Despite the fact that this assay is validated for serum samples, specimens of other body fluids are increasingly used for detection of galactomannan, including urine, bronchoalveolar lavage fluid, and cerebrospinal fluid. Review of the literature shows that galactomannan can be detected in each of these samples from patients with invasive aspergillosis with higher sensitivity than is the case with culture, as well as early in the course of infection. However, the evidence thus far is based on case reports--predominantly retrospective studies--that often include heterogeneous patient populations and limited numbers of cases of proven infection. Clearly, well-designed prospective studies with systematic sampling and use of consensus case definitions are needed to compare the performance of antigen detection in samples other than serum specimens with that in serum specimens. PMID- 15546084 TI - Development and experience with an algorithm to evaluate suspected smallpox cases in the United States, 2002-2004. AB - Concerns that smallpox, an eradicated disease, might reappear because of a bioterror attack and limited experience with smallpox diagnosis in the United States prompted us to design a clinical algorithm. We used clinical features of classic smallpox to classify persons presenting with suspected smallpox rashes into 3 categories: those with high, those with moderate, and those with low risk of having smallpox. The classification guides subsequent diagnostic strategies, limiting smallpox laboratory testing to high-risk persons to minimize the number of false-positive test results. From January 2002 through June 2004, the Centers for Disease Control and Prevention (CDC) received 43 consultations regarding suspected smallpox cases. No patient was at high risk for having smallpox. One patient was tested for the presence of variola virus. Varicella was the diagnosis for 23 cases (53%). The algorithm worked well to guide clinical and public health responses to suspected smallpox cases. The poster is available from CDC, and an interactive version and laboratory protocol are available at http://www.bt.cdc.gov/agent/smallpox/diagnosis/riskalgorithm/index.asp. We recommend use of the algorithm in the United States and elsewhere. PMID- 15546085 TI - Raccoon roundworm encephalitis. AB - The raccoon roundworm, Baylisascaris procyonis, is increasingly recognized as a cause of zoonotic visceral, ocular, and neural larva migrans and, in particular, of devastating encephalitis in young children. Exposure occurs mainly at raccoon latrines, where large numbers of infective eggs may be accidentally ingested. Risk factors for infection include contact with raccoon latrines, pica/geophagia, age of <4 years, and male sex. The severity of central nervous system (CNS) disease depends on the number of eggs ingested, the extent and location of larval migration, and the severity of ensuing inflammation and necrosis. Diagnosis of Baylisascaris encephalitis is based on clinical CNS disease, peripheral and cerebrospinal fluid eosinophilia, deep white matter lesions visible by magnetic resonance imaging, and positive results of serologic tests. Treatment efficacy in clinical cases is poor, but albendazole prevents disease if given promptly after infection. Considering the seriousness of this disease and limitations of diagnosis and treatment, prevention of infection with eggs is of utmost importance. PMID- 15546086 TI - Rickettsioses and the international traveler. AB - The rickettsioses--zoonotic bacterial infections transmitted to humans by arthropods--were for many years considered to be oddities in travel medicine. During the previous 2 decades, however, reports of >450 travel-associated cases have been published worldwide, the vast majority being murine typhus caused by Rickettsia typhi, Mediterranean spotted fever caused by Rickettsia conorii, African tick bite fever caused by Rickettsia africae, and scrub typhus caused by Orientia tsutsugamushi. Most patients present with a benign febrile illness accompanied by headache, myalgia, and cutaneous eruptions, but severe complications and fatalities are occasionally seen. Current microbiological tests include culture, polymerase chain reaction, and serological analysis, of which only the latter method is widely available. Tetracyclines are the drugs of first choice and should be prescribed whenever a case of rickettsiosis is suspected. Preventive measures rely on minimizing the risk of arthropod bites when traveling in areas of endemicity. PMID- 15546087 TI - Long-term follow-up of HIV-infected individuals who have significant increases in CD4+ cell counts during antiretroviral therapy. AB - BACKGROUND: Descriptions of the durability and consequences of immune reconstitution in patients who start highly active antiretroviral therapy (HAART) while severely immunosuppressed are limited. METHODS: Patients with previous CD4+ cell counts <50 cells/mm3, all of whom had HAART-induced increases in CD4+ cell counts of >100 cells/mm3 on 2 separate occasions (measured sequentially at least 4 weeks apart), were enrolled in a prospective trial and observed every 16-32 weeks. Evaluations included assessments for new opportunistic complications, virologic (human immunodeficiency virus [HIV] RNA load) and immunologic (CD4+ cell count) responses, or death. RESULTS: The median follow-up duration for 612 subjects was 184 weeks (range, 8-216 weeks). The rate of increase in CD4+ cell counts was approximately 5.9 cells/mm3 every 8 weeks, with the degree of increase associated with the baseline HIV RNA load (<500 vs. > or =500 copies/mL). Subsequent measurements of virologic suppression based on HIV RNA levels were also associated with predicted CD4+ cell responses. Thirty-three AIDS-defining illnesses were reported (1.75 events per 100 person-years of follow-up); >40% (14 cases) occurred with higher than expected CD4+ cell counts. CONCLUSIONS: CD4+ cell count increases are related to virological control, with continuing increases seen in individuals who are immunosuppressed. Opportunistic illnesses and/or complications are infrequent but can occur at any time, even in patients who maintained an elevated CD4+ cell count. PMID- 15546088 TI - Hepatitis C virus coinfection increases mortality in HIV-infected patients in the highly active antiretroviral therapy era: data from the HIV Atlanta VA Cohort Study. AB - BACKGROUND: We compared survival among patients coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) with that among patients infected solely with HIV. METHODS: Descriptive, bivariate, and survival analyses were conducted using data for all HIV-positive patients who were seen during the period of January 1997 through May 2001 in the HIV Atlanta VA Cohort Study (HAVACS) and who had been tested for HCV antibody since 1992 (n=970). RESULTS: The prevalence of HCV coinfection was 31.6%, and coinfected patients were significantly more likely to be older, black, and injection drug users. In multivariate analysis, the duration of survival from the time of diagnosis of acquired immunodeficiency syndrome (AIDS) was significantly shortened for HIV-HCV coinfected patients (hazard ratio [HR], 1.84; 95% confidence interval [CI], 1.09 3.10), as was time from HIV diagnosis to death (HR, 2.47; 95% CI, 1.26-4.82). Recovery of CD4+ cell count from the time of initiation of HAART did not differ significantly by coinfection status. CONCLUSIONS: HCV coinfection is common in this HIV-infected population and negatively affects survival from the time of both HIV and AIDS diagnoses, although this is apparently not associated with a difference in CD4+ cell recovery while receiving HAART. These findings differ from those of a previous study that was conducted in this cohort in the pre-HAART era, which found no association between HIV-HCV coinfection and HIV disease progression. PMID- 15546089 TI - Gynecomastia among HIV-infected patients is associated with hypogonadism: a case control study. AB - BACKGROUND: The prevalence, risk factors, and potential hormonal abnormalities associated with gynecomastia in a cohort of HIV-infected men are poorly understood. METHODS: Breast enlargement was assessed in consecutively evaluated HIV-infected men, and gynecomastia was subsequently confirmed with sonography. For each patient with breast enlargement, a randomly selected control subject without breast enlargement was studied. Clinical data were obtained, including age, body mass index, clinically evident lipodystrophy, prior symptomatic hyperlactatemia, current antiretroviral therapy and duration of exposure to each antiretroviral drug, history of injection drug use, and serological status regarding hepatitis B and hepatitis C. Laboratory parameters, including plasma HIV-1 RNA load, CD4 cell count, free testosterone index, and levels of fasting triglycerides, cholesterol, prolactin, total testosterone, sex hormone-binding globulin, 17-beta-estradiol, follicle-stimulating hormone, luteinizing hormone, and thyroid-stimulating hormone, were measured. RESULTS: There were 44 of 2275 patients with breast enlargement, of whom 40 (1.8%) had gynecomastia. The mean free testosterone index (+/-SD) was significantly lower among the 40 patients with gynecomastia (42.6%+/-24.0%) than among the 44 control subjects (58.0%+/ 25.3%) (P=.006). Although the proportion of patients who were receiving treatment with zidovudine, stavudine, and/or efavirenz at the time of the present study was significantly different between case patients and control subjects, the duration of exposure to each individual antiretroviral drug was not. Lipoatrophy (adjusted odds ratio [OR], 5.6; 95% confidence interval [CI], 1.7-18.6; P=.005), hepatitis C (adjusted OR, 6.1; 95% CI, 1.8-20.6; P=.003), and hypogonadism (adjusted OR, 7.6; 95% CI, 1.8-32.2; P=.003) were independent factors associated with gynecomastia. CONCLUSIONS: The data suggest that gynecomastia among HIV-infected patients is related to hypogonadism, rather than to an adverse effect of antiretroviral drugs. PMID- 15546090 TI - Oral miltefosine for leishmaniasis in immunocompromised patients: compassionate use in 39 patients with HIV infection. AB - Oral miltefosine was administered to 39 human immunodeficiency virus (HIV) infected patients with leishmaniasis for whom standard leishmaniasis treatment had failed. Initial response was achieved in 25 patients (64%), including 16 patients (43%) with initial parasitological cure. Repeated responses after relapse and tolerability of long courses of treatment indicate the potential for development of optimized dosage schemes. PMID- 15546091 TI - Native valve endocarditis due to coagulase-negative staphylococci: report of 99 episodes from the International Collaboration on Endocarditis Merged Database. AB - Using a large cohort of patients from the International Collaboration on Endocarditis Merged Database, we compared coagulase-negative staphylococcal (CoNS) native-valve endocarditis (NVE) to NVE caused by more common pathogens. Rates of heart failure and mortality were similar between patients with CoNS NVE and patients with Staphylococcus aureus NVE, but rates for both groups were significantly higher than rates for patients with NVE due to viridans streptococci. These results emphasize the importance of CoNS as a cause of NVE and the potential for serious complications with this infection. PMID- 15546092 TI - Inhalation of nitric oxide in the treatment of severe acute respiratory syndrome: a rescue trial in Beijing. AB - Inhalation of nitric oxide (NO) improved arterial oxygenation and enabled the reduction of inspired oxygen therapy and airway pressure support in patients with severe acute respiratory syndrome (SARS). In addition, chest radiography showed decreased spread or density of lung infiltrates, and the physiological effects remained after termination of inhaled NO therapy. These findings suggest not only a pulmonary vasodilator effect of inhaled NO, but also an effect on SARS. PMID- 15546093 TI - Association between tea ingestion and invasive Bacillus cereus infection among children with cancer. AB - Bacillus cereus is an emerging pathogen that causes invasive disease in immunocompromised hosts. A case-control study, prompted by a clinical case, demonstrated an association between dietary tea ingestion and B. cereus bacteremia. Policies designed to interrupt transmission of this pathogen to susceptible patients should be considered. PMID- 15546096 TI - Failure of vancomycin for treatment of methicillin-resistant Staphylococcus aureus infections. PMID- 15546094 TI - Infrequent reinfection after successful treatment for hepatitis C virus infection in injection drug users. AB - We followed-up 18 injection drug users for a mean of 33.8 months (range, 4-55 months) after successful treatment for hepatitis C virus (HCV) infection. Fifteen (83%) of the patients remained HCV RNA-negative, 1 patient was not tested, and 2 patients had test results positive for HCV RNA. The estimated rate of reinfection as a result of injection drug use was 0-4.1 cases per 100 person-years (cumulative incidence, 0%-12.6% at 48 months after completion of treatment). Of 50 patients originally treated, 15 (30%) were HCV RNA-negative 3 years later. PMID- 15546097 TI - Treating HIV encephalopathy with antiretroviral therapy: a clinical case demonstrating the success of HAART. PMID- 15546098 TI - Fatal septicemia and pyomyositis caused by Salmonella typhi. PMID- 15546099 TI - Impact of antiretroviral therapy among HIV-1-infected patients with pulmonary hypertension. PMID- 15546102 TI - Chronic liver disease and Streptococcus bovis endocarditis. PMID- 15546103 TI - Furuncular myiasis after contact with clothing (when washing clothes can be infectious). PMID- 15546104 TI - HIV-2: the Portuguese connection. PMID- 15546106 TI - Fluoroquinolone-resistant Streptococcus pneumoniae, an emerging but unrecognized public health concern: is it time to resight the goalposts? PMID- 15546108 TI - Management of community-acquired respiratory tract infections. PMID- 15546109 TI - Increased antibiotic resistance in respiratory tract pathogens: PROTEKT US--an update. AB - Three major North American surveillance programs have tracked antimicrobial resistance patterns among isolates of Streptococcus pneumoniae and other common respiratory tract pathogens. The Canadian Bacterial Surveillance Network shows the progressive increase in resistance among pneumococcal S. pneumoniae to penicillin, trimethoprim-sulfamethoxazole, macrolides, and fluoroquinolones. The data from the Tracking Resistance in the United States Today study also show a steady rise in pneumococcal resistance among common antibiotics as well as an increase in multidrug-resistant S. pneumoniae. The US component of the Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin study has detected increasing resistance to many antimicrobial agents among common respiratory isolates, with marked geographic variations in resistance patterns. The patterns of resistance detected by these major surveillance programs are a warning signal regarding the continued emergence of resistance among community-acquired respiratory tract pathogens. PMID- 15546110 TI - Acute community-acquired bacterial sinusitis: continuing challenges and current management. AB - Acute sinusitis is one of the most common infections seen in general clinical practice. The most common cause of acute sinusitis is viral; however, many patients receive a prescription for an antibiotic. Such injudicious prescribing habits have a major impact on health care costs, contribute to the increasing prevalence of drug-resistant strains of common respiratory pathogens, and reflect many of the challenges in differentiating viral and bacterial disease. Sinus puncture and culture of the aspirate, the diagnostic reference standard in the research setting, are not appropriate for routine clinical practice. However, certain clinical signs and symptoms that do not improve or that worsen after 7-10 days are currently accepted criteria for diagnosis of bacterial sinusitis. Accurate diagnosis can select patients who would benefit most from antimicrobial use. Antimicrobial agents should be selected on the basis of local resistance patterns, and their spectrum of activity should cover the common bacterial pathogens, including resistant strains. PMID- 15546111 TI - Clinical efficacy of newer agents in short-duration therapy for community acquired pneumonia. AB - Streptococcus pneumoniae, the most important respiratory tract pathogen implicated in community-acquired pneumonia (CAP), is becoming increasingly resistant in vitro to the beta -lactams and macrolides, and fluoroquinolone resistance has been detected. A growing body of evidence suggests that prolonged antimicrobial use may contribute directly and indirectly to increased antimicrobial resistance among common respiratory pathogens. Long-term exposure to antimicrobial agents, especially less-potent agents, directly increases selection pressure for resistance. Indirectly, poor patient compliance, multiple daily dosing, and the increased risk of adverse events further complicate the resistance issue and diminish the efficacy of long-term antimicrobial use. Controlled clinical trials addressing the appropriate duration of therapy for CAP are lacking. However, available data suggest that with appropriate antibiotic selection, based on appropriate spectrum, potency, and pharmacokinetic/pharmacodynamic profile, lower respiratory tract infections in outpatients can be successfully treated in <7 days rather than the 7-14 days currently recommended. PMID- 15546112 TI - The design of clinical trials that evaluate antifungal prophylaxis and combination therapy: introduction and overview. AB - On 8 February 2003, the second in a series of meetings in honor of John E. Bennett, M.D., was convened in New York City. A report from the previous meeting was published in 2003. The goal of this second meeting was to discuss the design of clinical trials in prophylaxis and combination therapy. This supplement presents 12 articles by leading clinicians who are currently active in trials in this area and presents a current view of the unmet needs and challenges. PMID- 15546113 TI - Issues in the design of the fluconazole prophylaxis trials in patients undergoing hematopoietic stem cell transplantation. AB - Today, invasive candidal infections are much less of a problem in patients undergoing hematopoietic stem cell transplantation (HSCT) than they were in the 1980s, principally because of widespread application of effective antifungal prophylaxis strategies. Two randomized, placebo-controlled studies performed in the early 1990s were instrumental in demonstrating the protective effects of fluconazole. Although both studies showed that candidiasis is prevented by fluconazole prophylaxis, controversies regarding optimal administration guidelines and the impact on survival after HSCT have persisted over the last decade. Details of how these trials were done, including the patients enrolled and end points evaluated, explain many of the different findings and controversies. An understanding of design issues can help explain current controversies and can assist in development of future prophylaxis studies. PMID- 15546114 TI - Design issues in a prospective randomized double-blinded trial of prophylaxis with fluconazole versus voriconazole after allogeneic hematopoietic cell transplantation. AB - BACKGROUND: Aspergillus infections pose the toughest infectious challenges to the clinician caring for hematopoietic cell transplant recipients. About 15% of patients become infected, with a case fatality rate of approximately 65%. To date, no effective prophylactic strategies have been developed. METHODS: Voriconazole, a recently licensed extended-spectrum azole, with demonstrated efficacy against aspergillus, is currently being tested as a potential prophylactic agent against aspergillus and other invasive fungal infections. Logistic issues--such as patient selection, choice of comparator, blinding of study drugs, duration of study drug administration, and how to handle empirical amphotericin B for possible invasive fungal infections--and analytic concerns, including choice and definition of the primary end point and the potential confounding effect of informative censoring (as a result of noninfectious events), were considered in the design of the clinical trial. RESULTS: The trial is now under way, with a projected 3-year enrollment period. CONCLUSIONS: Each design decision shaped the trial in a way that permitted certain questions to be answered while not allowing others to be addressed. Once completed, the trial's results must be interpreted in light of these design details. PMID- 15546115 TI - How to improve the design of trials of antifungal prophylaxis among neutropenic adults with acute leukemia. AB - The risk for invasive fungal infections in patients with acute leukemia is generally low (4%-8%), and the routine use of fungal prophylaxis is not warranted except in specific high-risk groups that should be identified among this population. In a prophylactic study with a new agent, fluconazole or itraconazole oral solution represent good choices for the comparator because they are proven better than placebo or oral nonabsorbable antifungal agents in reducing the risk of invasive fungal infections in patients with acute leukemia. Because prophylaxis is most valuable when the risk of infection is high, patients with well-understood risk factors (severe mucosal disruption caused by chemotherapy, impaired cell-mediated immunity caused by steroids or fludarabine, use of a central venous catheter, and colonization by Candida species) should be selected. The end points for antifungal prophylactic trials should focus on proven and probable invasive fungal infections. Superficial and mucosal fungal infections do not represent a primary end point for these studies. Poor compliance should be considered as an interruption of treatment due to side effects and should be included in the criteria for failure. Fungus-related mortality should be evaluated as a failure of prophylaxis, whereas overall mortality may be influenced by many other cofactors. Differences in gastrointestinal toxicity of antifungal agents may limit the use of double-blind designs in some situations. PMID- 15546116 TI - Clinical trials of antifungal prophylaxis among patients undergoing surgery. AB - Invasive mycoses have emerged as a major cause of morbidity and mortality. Epidemiological studies have shown that surgery services have the highest rate of Candida infections in the hospital. In addition to classical risk factors, heavy Candida colonization, recurrent gastrointestinal perforations, and acute pancreatitis are frequently associated with invasive candidiasis. Because prompt initiation of antifungal therapy is critical for cure but difficult to accomplish, prevention of fungal infections may play an important role in this clinical setting; however, few prophylactic or preemptive studies have been done to date. The choice, route of administration, and dose of the antifungal and comparator regimens and the use of clinically relevant and robust study end points are critical for the trial design. Various criteria have been used to identify patients at risk of candidiasis: surgical condition, presence of multiple risk factors, colonization indexes, or expected length of stay in the intensive care unit. Some are not selective enough, and others are time consuming and expensive. Rigorous selection of high-risk patients is crucial to optimize the risk-benefit ratio of preventive antifungal strategies. The aim is to maximize chances of reducing morbidity and mortality while minimizing treatment costs, exposure of low-risk patients to adverse events, and emergence of resistant fungal strains. PMID- 15546117 TI - Clinical trials of antifungal prophylaxis among patients in surgical intensive care units: concepts and considerations. AB - BACKGROUND: Fungal infections are important clinical infections in patients in surgical intensive care units. In some institutions, antifungal prophylaxis has become commonplace, and increasing resistance has been reported. However, trials of antifungal prophylaxis are hampered by difficulties in trial design, and the findings may not be generalizable. METHODS: Issues in clinical trial design are reviewed from existing and theoretical perspectives. RESULTS: Identification of a primary hypothesis with a sound epidemiological basis is essential. The study must include institutions where fungal infections have a high and well-studied incidence. A high-risk patient population should be identified and enrolled. The agent selected should have an appropriate spectrum, be easily delivered to the population selected, and be cost effective with few adverse events. At present, fluconazole appears to be the best agent for targeted prophylaxis. The primary end point of the study should be based on an easily measured outcome, for example, days free from fungal infection rather than death due to fungal infection. CONCLUSIONS: Trials of antifungal prophylaxis for patients in surgical intensive care units have had problems in design, and several issues in the conceptual basis of future clinical trials must be addressed. PMID- 15546118 TI - Antifungal prophylaxis in solid-organ transplant recipients: considerations for clinical trial design. AB - BACKGROUND: There are several key issues regarding clinical trial design for antifungal prophylaxis in organ transplant recipients. METHODS: The existing and emerging data on the epidemiology and risk factors for invasive mycoses in organ transplant recipients formed the basis for discerning the subgroups that may be targeted for prophylaxis, the unique end points to be considered, and the attributes of currently available drugs that may be suitable candidates for antifungal prophylaxis. RESULTS: Epidemiological characteristics of invasive mycoses in organ transplant recipients have evolved sufficiently to warrant thoughtful reconsideration of the subsets of patients and the fungal pathogens to be targeted for antifungal prophylactic trials in the current era. Safety and tolerability of antifungal agents and outcome stratified by severity of illness are important end points given the potential for drug interactions with immunosuppressive agents and the fact that a beneficial effect of prophylaxis on outcome has been difficult to document in organ transplant recipients. CONCLUSIONS: Clinical trial design for antifungal prophylaxis must carefully consider the unique issues pertaining to the selection of patients most likely to benefit, as well as the tolerability and drug interactions of antifungal agents in organ transplant recipients. PMID- 15546119 TI - Design of aerosolized amphotericin b formulations for prophylaxis trials among lung transplant recipients. AB - BACKGROUND: Emphasis has been placed on the need for antifungal prophylaxis when transplanted organs are directly exposed to the environment. In lung transplantation, one strategy for prevention has been aerosolization of polyenes into the lung. Despite widespread use of amphotericin B aerosols, there are still few studies on how to validate their effectiveness or optimize pharmacokinetics. METHODS: We compared results of several of our own studies of lung transplant recipients receiving formulations of amphotericin B with results of other studies. RESULTS: Recent aerosol studies suggest that aerosols of a lipid formulation of amphotericin B are safe and better tolerated than is amphotericin B deoxycholate. Their use in prophylaxis is associated with very few pulmonary fungal infections. CONCLUSION: This safe drug delivery at a local body site to directly protect the transplanted organ without systemic antifungal drug exposure is attractive from a cost and toxicity standpoint. However, careful multicenter, comparative studies are still needed to ensure that this strategy is consistently successful. PMID- 15546120 TI - Issues in clinical trials of prophylaxis of fungal infections. AB - The validity of the results of a clinical trial is highly dependent upon the design of the trial. The definition of disease, the selection criteria for enrollment in the trial, the selection of the study and control drugs, and the end points all affect whether the information obtained from the trial ultimately is useful in making decisions in clinical practice. These factors all apply to the design of clinical trials of the prophylaxis of infectious diseases. In addition, prophylaxis trials have several important differences from the design of trials of the treatment of those same diseases. The risk-benefit analysis for trials of prophylaxis is different, in that asymptomatic patients are exposed to the drug and more patients will be exposed than will develop the disease under study. Standardization of the design of such clinical trials will allow more efficient development of new drugs and will allow clinicians to compare more accurately the safety and efficacy of prophylactic agents. PMID- 15546121 TI - Design of efficacy trials of cytokines in combination with antifungal drugs. AB - Resolution of invasive fungal infections is often dependent on recovery from an immunocompromised state, which indicates that host defense mechanisms are extremely important in the clearance of fungal pathogens. Immunotherapy aimed at enhancement of host defense mechanisms may improve clinical outcome of invasive mycoses. The design of trials of immunotherapy against fungal pathogens requires profound knowledge of the host defense mechanisms that are involved in invasive fungal infections. Prospective phase II studies with recombinant granulocyte colony-stimulating factor and interferon-gamma have been done. Recombinant interferon-gamma is a candidate for phase III trials of adjunctive immunotherapy for cryptococcal meningitis, invasive aspergillosis, and candidemia, but the proper design of future trials will be crucial to establish whether immunotherapy is of clinical value in the treatment of invasive fungal infections. PMID- 15546122 TI - Combination therapy for invasive mycoses: evaluation of past clinical trial designs. AB - BACKGROUND: Despite the availability of new antifungals, single-agent therapy frequently falls short of high cure rates. Combination therapy offers potentially higher cure rates, especially for resistant organisms. In vitro studies and experimental animal models have provided conflicting data. METHODS: Retrospective, randomized, controlled clinical studies were reviewed. RESULTS: Results indicate a clear advantage for polyene and flucytosine combination therapy in cryptococcal meningitis and a possible advantage for combination amphotericin B and fluconazole for candidemia. Unfortunately, the few studies published have been flawed by design problems that have compromised the determination of outcome. Study review allows investigators the opportunity to design future studies to ensure optimal evaluation of efficacy. CONCLUSION: Combination antifungal therapy is advantageous in cryptococcal diseases and may have a role in the treatment of invasive candidiasis. The greatest potential exists for combination therapy against aspergillosis and resistant fungi in patients with refractory mycotic disease who experience failure of monotherapy. PMID- 15546123 TI - Considerations in clinical trials of combination antifungal therapy. AB - The cure rate for serious fungal diseases with currently available agents used as monotherapy is not optimal. The introduction of new classes of antifungal drugs in the last few years naturally leads to the hypothesis that antifungal drugs used in combination may be more effective than the same drugs used alone. The design and interpretation of combination therapy studies raise challenges beyond those encountered when drugs are studied as monotherapy in the treatment of a disease. The definition of combination therapy, the study design, the selection of appropriate patient populations, and the selection of end points, as well as practical considerations, are all important in the design and interpretation of clinical trials of combination therapies. Although combination therapies hold the promise of improved efficacy, it is important to prove this hypothesis, because they also may be associated with increased toxicity and increased drug costs. A careful consideration of study design factors before the initiation of a trial will help obtain the most useful information for patients in this important area. PMID- 15546124 TI - The validity of the family relationships index as a screening tool for psychological risk in families of cancer patients. AB - The Family Relationships Inventory (FRI) has been proposed as a measure of psychological risk however its validity has only been tested in one cross sectional study against another measure of family functioning where patients had less than six months to live. The current study presented longitudinal data on the validity of the FRI in identifying family dysfunction, and clinical levels of depression and anxiety in 48 families where the patient had been recently diagnosed. Over the three phases, the FRI identified all families at risk of family dysfunction, 88% or more families with one or more members with clinical depression and 78% or more with a member with clinical levels of anxiety. The FRI was also far more sensitive in identifying families with a member with clinical levels of depression and anxiety than the General Functioning (FAD-GF) scale of the Family Assessment Device. Although other measures of screening adequacy (such as specificity), suggested that the FRI identified too many false positives, the higher sensitivity of the FRI makes it preferable to the FAD-GF as a screening measure for families at risk of a poor psychological outcome in the oncology setting. However, because of the FRI's poor specificity, further follow-up of those families that are identified as being at some risk of a poor psychological outcome should be undertaken before referral to a mental health professional is warranted. PMID- 15546125 TI - Posttranslational activation, site-directed mutation and phylogenetic analyses of the lysine biosynthesis enzymes alpha-aminoadipate reductase Lys1p (AAR) and the phosphopantetheinyl transferase Lys7p (PPTase) from Schizosaccharomyces pombe. AB - Alpha-aminoadipate reductase (AAR), the signature enzyme for lysine biosynthesis in fungi, catalyses the conversion of alpha-aminoadipate to alpha-aminoadipate semiadehyde in the presence of ATP and NADPH. In Saccharomyces cerevisiae and Candida albicans, the LYS2-encoded AAR is posttranslationally activated by CoA and the LYS5-encoded PPTase. The fission yeast Schizosaccharomyces pombe is evolutionarily highly diverged from S. cerevisiae and C. albicans. We report here several unusual activation characteristics of Sz. pombe Lys1p and Lys7p, isofunctional to Lys2p (AAR) and Lys5p (PPTase), respectively. Unlike the Lys2p from S. cerevisiae and C. albicans, the Sz. pombe Lys1p was active when expressed in E. coli and exhibited significant AAR activity without the addition of CoA or the Sz. pombe Lys7p intron free PPTase. Somewhat higher AAR activity was obtained with the addition of CoA and the Sz. pombe Lys7p PPTase. Substitution of G910A, S913T or S913A in the Sz. pombe Lys1p activation domain (IGGHSI) resulted in no AAR activity. Similarly, substitutions of several amino acid residues in the Sz. pombe Lys7p PPTase domain (G79A, R80K and P81A in Core 1; F93W, D94E, F95W and N96D in Core 1a; G124A, V125I and D126E in Core 2; K172R, E173D and K177R in Core 3) also resulted in no activation of Lys1p and no AAR activity. The Sz. pombe Lys1p amino acid sequence showed a high degree of similarity to other fungal Lys2p proteins; however, the Lys7p amino acid sequence showed much less similarity to other bacterial, fungal and animal PPTases representing several phylogenetic groups. PMID- 15546126 TI - Endothelin receptor expression in human lungs of newborns with congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a major cause of refractory respiratory failure in the neonatal period and is characterized by persistent pulmonary hypertension of the newborn (PPHN) and pulmonary hypoplasia. Endothelin-1 (ET-1) dysregulation may play a significant role in the pathophysiology of PPHN and ET-1 acts through binding to type A (ETA) and type B (ETB) receptors. Therefore, ETA and ETB receptor protein expression was studied using immunohistochemistry in 10 lung specimens obtained from newborns with CDH, and 4 normal lung specimens, in order to explore whether dysregulation of ETA and ETB expression contributes to PPHN. ETA and ETB mRNAs were then quantified using real-time RT-PCR in laser microdissected pulmonary resistive arteries. In the lungs of newborns with CDH, immunohistochemistry of both ETA and ETB receptors demonstrated over-expression in the thickened media of pulmonary arteries. Using laser microdissection and real-time RT-PCR, higher levels of ETA and ETB mRNA were found in CDH pulmonary arteries than in controls: this increase was more pronounced for ETA mRNA. This study provides the first demonstration of ET-1 receptor dysregulation in association with structural alteration of pulmonary arteries in newborns with CDH and PPHN. This dysregulation preferentially affects the ETA receptor. These results suggest that dysregulation of ET-1 receptors may contribute to PPHN associated with CDH. PMID- 15546127 TI - Measles as a potential oncolytic virus. AB - The use of replicating viruses for cancer therapy is attracting increasing interest. Numerous viruses are now being considered as potential cancer therapeutics, including the vaccine strain of measles virus (MV). The attenuated strain of measles readily lyses transformed cells, whilst replication and lysis are limited in normal human cells. It has a number of features which make it highly suitable for further development as an oncolytic agent, among them stability and a long history of safety in human use. These features are being combined with its ready potential for genetic manipulations to generate recombinant MVs with desirable therapeutic attributes. This review summarises the pre-clinical studies of the oncolytic efficacy of MV to date. Promising developments in MV engineering--such as re-targeting MV entry to specific cell types and enhancing its utility as a therapeutic agent by expression of non-viral proteins--as well as outstanding issues, such as the role of anti-MV immunity, are highlighted. PMID- 15546128 TI - Reactivation of Epstein-Barr virus from latency. AB - The general problem in cancer treatment centres on finding agents that specifically affect cancer cells without damaging normal cells. The differences between cancer cells and normal cells are usually very subtle but about 15% of all human cancers involve a virus infection, for example the Epstein-Barr virus associated cancers. In these cancers, every tumour cell carries the virus in a latent infection but the number of normal cells infected is very low. So a treatment that could somehow cause the elimination of EBV infected cells would be very specific for the cancer in such cases. One potential approach could involve finding ways to reactivate the latent virus in cancer cells into the early part of the lytic cycle, impeding cell proliferation, targeting chemotherapeutic agents to the cancer and causing the cancer cells to become targets for immune surveillance. This review considers the mechanisms by which EBV reactivation is controlled and discusses possible therapeutic approaches. PMID- 15546129 TI - Recombination in alphaherpesviruses. AB - Within the Herpesviridae family, Alphaherpesvirinae is an extensive subfamily which contains numerous mammalian and avian viruses. Given the low rate of herpesvirus nucleotide substitution, recombination can be seen as an essential evolutionary driving force although it is likely underestimated. Recombination in alphaherpesviruses is intimately linked to DNA replication. Both viral and cellular proteins participate in this recombination-dependent replication. The presence of inverted repeats in the alphaherpesvirus genomes allows segment inversion as a consequence of specific recombination between repeated sequences during DNA replication. High molecular weight intermediates of replication, called concatemers, are the site of early recombination events. The analysis of concatemers from cells coinfected by two distinguishable alphaherpesviruses provides an efficient tool to study recombination without the bias introduced by invisible or non-viable recombinants, and by dominance of a virus over recombinants. Intraspecific recombination frequently occurs between strains of the same alphaherpesvirus species. Interspecific recombination depends on enough sequence similarity to enable recombination between distinct alphaherpesvirus species. The most important prerequisite for successful recombination is coinfection of the individual host by different virus strains or species. Consequently the following factors affecting the distribution of different viruses to shared target cells need to be considered: dose of inoculated virus, time interval between inoculation of the first and the second virus, distance between the marker mutations, genetic homology, virulence and latency. Recombination, by exchanging genomic segments, may modify the virulence of alphaherpesviruses. It must be carefully assessed for the biosafety of antiviral therapy, alphaherpesvirus-based vectors and live attenuated vaccines. PMID- 15546131 TI - Non-empirical calculations of NMR indirect carbon-carbon coupling constants. Part 11--saturated carbocycles: a reference data set and a practical guide to structural elucidation. AB - Carbon-carbon spin-spin coupling constants, J(C,C), calculated at the SOPPA level for 50 mono-, bi- and polycycloalkanes in 10 previous papers are put in order and discussed on unified grounds. Basic structural trends of J(C,C) established in the original publications are summarized and briefly outlined for the representative series. Many unknown couplings are predicted with high reliability, and this provides a good reference data set and a practical guide to the structural elucidation of saturated carbocycles by means of J(C,C) coupling constants. PMID- 15546130 TI - Factors affecting delivery of antiviral drugs to the brain. AB - Although the CNS is in part protected from peripheral insults by the blood-brain barrier and the blood-cerebrospinal fluid barrier, a number of human viruses gain access to the brain, replicate within this organ, or sustain latent infection. The efficacy of antiviral drugs towards the cerebral viral load is often limited as both blood-brain interfaces impede their cerebral distribution. For polar compounds, the major factor restricting their entry lies in the tight junctions that occlude the paracellular pathway across these barriers. For compounds with more favourable lipid solubility properties, CNS penetration will be function of a number of physicochemical factors that include the degree of lipophilicity, size and ability to bind to protein or red blood cells, as well as other factors inherent to the vascular and choroidal systems, such as the local cerebral blood flow and the surface area available for exchange. In addition, influx and efflux transport systems, or metabolic processes active in both capillary endothelial cells and choroid plexus epithelial cells, can greatly change the bioavailability of a drug in one or several compartments of the CNS. The relative importance of these various factors with respect to the CNS delivery of the different classes of antiviral drugs is illustrated and discussed. PMID- 15546132 TI - Measurement error model for misclassified binary responses. AB - The article considers regression models for binary response in a situation when the response is subject to classification error. It is also assumed that some of the covariates are unobservable, but measurements on its surrogates are available. Likelihood based analysis is developed to fit the model. A sensitivity analysis is also carried out through simulation to ascertain the effect of ignoring classification error and/or measurement error on the estimation of regression parameters. At the end, the methodology developed in this paper is illustrated through an example. PMID- 15546133 TI - Overlap bias in the case-crossover design, with application to air pollution exposures. AB - The case-crossover design uses cases only, and compares exposures just prior to the event times to exposures at comparable control, or 'referent' times, in order to assess the effect of short-term exposure on the risk of a rare event. It has commonly been used to study the effect of air pollution on the risk of various adverse health events. Proper selection of referents is crucial, especially with air pollution exposures, which are shared, highly seasonal, and often have a long term time trend. Hence, careful referent selection is important to control for time-varying confounders, and in order to ensure that the distribution of exposure is constant across referent times, a key assumption of this method. Yet the referent strategy is important for a more basic reason: the conditional logistic regression estimating equations commonly used are biased when referents are not chosen a priori and are functions of the observed event times. We call this bias in the estimating equations overlap bias. In this paper, we propose a new taxonomy of referent selection strategies in order to emphasize their statistical properties. We give a derivation of overlap bias, explore its magnitude, and consider how the bias depends on properties of the exposure series. We conclude that the bias is usually small, though highly unpredictable, and easily avoided. PMID- 15546134 TI - Atypical protein kinase C-zeta modulates clonogenicity, motility, and secretion of proteolytic enzymes in murine mammary cells. AB - In this paper, we investigated whether protein kinase C-zeta (PKC zeta), a member of the atypical PKC family, induces phenotypic alterations associated with malignant transformation and tumor progression in mammary cells. The stable overexpression of PKC zeta in immortalized mammary epithelial cells (NMuMG), activates the mitogenic extracellular signal-regulated kinase (ERK) pathway, enhanced clonal cell growth and exerts profound effects on proteases secretion. The effect on proteases expression seems to be specific for urokinase-type plasminogen activator and metalloproteinase-9 (MMP-9) because no modulation in MMP-2 and MMP-3 production could be detected. In addition, our experiments demonstrated that PKC zeta overexpression markedly altered the adhesive, spreading, and migratory abilities of NMuMG cells. The overexpression of this enzyme was not sufficient to confer an anchorage-independent growth capacity. An extensive mutational analysis of PKC zeta revealed that the effects observed in NMuMG cells were strictly dependent on the kinase (catalytic) domain of the enzyme. Taken together, these results suggest that in mammary cells PKC zeta modulates several of the critical events involved in tumor development and dissemination through the activation of mitogen activated protein kinase (MAPK)/ERK pathway. PMID- 15546135 TI - High dose melphalan in the treatment of advanced neuroblastoma: results of a randomised trial (ENSG-1) by the European Neuroblastoma Study Group. AB - BACKGROUND: High dose myeloablative chemotherapy ("megatherapy"), with haematopoietic stem cell support, is now widely used to consolidate response to induction chemotherapy in patients with advanced neuroblastoma. PROCEDURE: In this study (European Neuroblastoma Study Group, ENSG1), the value of melphalan myeloablative "megatherapy" was evaluated in a randomised, multi-centre trial. Between 1982 and 1985, 167 children with stages IV and III neuroblastoma (123 stage IV > 1 year old at diagnosis and 44 stage III and stage IV from 6 to 12 months old at diagnosis) were treated with oncovin, cisplatin, epipodophyllotoxin, and cyclophosphamide (OPEC) induction chemotherapy every 3 weeks. After surgical excision of primary tumour, the 90 patients (69% of the total) who achieved complete response (CR) or good partial response (GPR) were eligible for randomisation either to high dose melphalan (180 mg per square meter) with autologous bone marrow support or to no further treatment. RESULTS: Sixty-five (72%) of eligible children were actually randomised and 21 of these patients were surviving at time of this analysis, with median follow-up from randomisation of 14.3 years. Five year event-free survival (EFS) was 38% (95% confidence interval (CI) 21-54%) in the melphalan-treated group and 27% (95% CI 12-42%) in the "no-melphalan" group. This difference was not statistically significant (P = 0.08, log rank test) but for the 48 randomised stage IV patients aged >1 year at diagnosis outcome was significantly better in the melphalan treated group-5 year EFS 33% versus 17% (P = 0.01, log rank test). CONCLUSIONS: In this trial, high dose melphalan improved the length of EFS and overall survival of children with stage IV neuroblastoma >1 year of age who achieved CR or GPR after OPEC induction therapy and surgery. Multi-agent myeloablative regimens are now widely used as consolidation therapy for children with stage IV disease and in those with other disease stages when the MYCN gene copy number in tumour cells is amplified. Because they are more toxic, complex, and costly these combination megatherapy regimens should be compared with single agent melphalan in randomised clinical trials. PMID- 15546136 TI - Interdependence of N-acetyl aspartate and high-energy phosphates in healthy human brain. AB - Because cellular and extract data have suggested that N-acetylaspartate (NAA) reflects neuronal mitochondrial function, we evaluated the quantitative relationship between NAA, high-energy phosphates, and ADP levels in the hippocampus and occipital lobe of 15 healthy volunteers. The ADP levels are calculated using the creatine kinase equilibrium and quantified (31)P and total creatine measurements. Using high-field quantitative MR spectroscopic imaging, we find that NAA and ADP concentrations in the hippocampal body are 9.7 +/- 1.5mM and 35 +/- 8microM, respectively. In the occipital lobe, NAA and ADP are 11.9 +/- 1.9mM and 32 +/- 12microM, respectively. There is a statistically significant positive correlation between NAA and ADP, with R = +0.80, p < 2 x 10(-7)in the hippocampal body. In an adjacent hippocampal NAA voxel, the correlation between NAA and ADP had a R = +0.62, p < 3 x 10(-4), whereas, in the occipital lobe, R = +0.67, p < 5 x 10(-5). There was no significant relationship NAA and either ATP or phosphocreatine. This positive relationship of NAA with ADP suggests a directional process wherein energetics may modulate mitochondrial function. PMID- 15546137 TI - Prognostic factors in oral cavity and oropharyngeal squamous cell carcinoma. AB - BACKGROUND: The survival of patients with head and neck squamous cell carcinoma (HNSCC) remains unaffected despite recent therapeutic advances. To reverse this trend, reliable and clinically applicable markers of tumor aggressiveness must be identified. One such marker may be the tumor-associated macrophage content. The authors hypothesized that tumor-associated macrophages contribute to HNSCC aggressiveness, and the objective of the current study was to prove this hypothesis using mRNA expression analysis and a large cohort of clinical specimens. METHODS: Oligonucleotide microarray analysis was performed on a prospective cohort of 20 patients with previously untreated oral cavity or oropharynx squamous cell carcinoma (OC/OP SCCA) and on normal oropharyngeal mucosa from 4 patients. After determining whether macrophage chemoattractants were produced by tumors, conditioned media from three HNSCC cell lines were used to quantify macrophage migration in an in vitro assay. A high-density tissue microarray of 102 patients with previously untreated OC/OP SCCA was stained immunohistochemically for CD68 to identify tissue macrophages, and the results were correlated with clinicopathologic data and survival. RESULTS: Monocyte chemoattractant protein 1 was up-regulated significantly in tumors compared with normal mucosa (P=0.0025; fold change=1.89). All University of Michigan SCC tumor cell line conditioned media caused a significant increase in macrophage migration (P <0.05). Tissue microarray data revealed that macrophage content of the primary tumor was associated strongly with lymph node metastasis (P <0.0001), extracapsular lymph node spread (P=0.0001), and advanced clinical disease stage (P=0.0002). When it was evaluated along with other clinicopathologic data, the macrophage content was found to be an independent predictor of lymph node metastasis (P <0.0001). CONCLUSIONS: Primary tumor macrophage content is a strong predictor of tumor aggressiveness in HNSCC. PMID- 15546138 TI - Sulindac and its metabolites inhibit invasion of glioblastoma cells via down regulation of Akt/PKB and MMP-2. AB - Non-steroidal anti-inflammatory drug (NSAID), sulindac has chemopreventive and anti-tumorigenic properties, however, the molecular mechanism of this inhibitory action has not been clearly defined. The Akt/protein kinase B, serine/threonine kinase is well known as an important mediator of many cell survival signaling pathways. In the present study, we demonstrate that down-regulation of Akt is a major effect of anti-invasiveness property of sulindac and its metabolites in glioblastoma cells. Myristoylated Akt (MyrAkt) transfected U87MG glioblastoma cells showed increase invasiveness, whereas DN-Akt transfected cells showed decrease invasiveness indicating that Akt potently promoted glioblastoma cell invasion. MMP-2 promoter and enzyme activity were up-regulated in Akt kinase activity dependent manner. Sulindac and its metabolites down-regulated Akt phosphorylation, inhibited MMP-2 production, and significantly inhibited invasiveness of human glioblastoma cells. In addition, sulindac and LY294002, a selective inhibitor of phosphoinositide 3-kinase (PI3K), synergistically inhibited the invasion of glioblastoma cells. Furthermore, only celecoxib showed Akt phosphorylation reduction and an anti-invasivness in glioblastoma cells, whereas aspirin, ketoprofen, ketorolac, and naproxen did not. In conclusion, our results provide evidence that down-regulation of Akt pathway and MMP-2 may be one of the mechanisms by which sulindac and its metabolites inhibit glioblastoma cell invasion. PMID- 15546139 TI - DNA methylation and chromatin structure: the puzzling CpG islands. AB - DNA methylation is the epigenetic modification, which introduces 5mC as fifth base onto DNA. As for the distribution of 5mCs, it is well known that they distribute themselves in a non-random fashion in genomic DNA so that methylation pattern is characterized by the presence of methylated cytosines on the bulk of DNA while the unmethylated ones are mainly located within particular regions termed CpG islands. These regions represent about 1% of genomic DNA and are generally found in the promoter region of housekeeping genes. Their unmethylated state, which is an essential condition for the correct expression of correlated genes, is paradoxical if one considers that these regions are termed CpG islands because they are particularly rich in this dinucleotide, which is the best substrate for enzymes involved in DNA methylation. Anomalous insertion of methyl groups in these regions generally leads to the lack of transcription of correlated genes. An interesting scientific problem is to clarify the mechanism(s) whereby CpG islands, which remain protected from methylation in normal cells, are susceptible to methylation in tumor cells. How the CpG moieties in CpG islands become vulnerable or resistant to the action of DNA methyltransferases and can thus lose or maintain their characteristic pattern of methylation is still an open question. Our aim is to gather some mechanisms regarding this intriguing enigma, which, despite all energy spent, still remains an unresolved puzzle. PMID- 15546140 TI - Stat1-mediated cytoplasmic attenuation in osteoimmunology. AB - Signal transducer and activator of transcription 1 (Stat1) is a critical mediator of gene transcription in type I interferon (IFN-alpha/beta) signaling that is essential for host defense against viruses. In the skeletal system, type I IFNs (IFN-alpha/beta) also play an important physiological role in the inhibition of receptor activator of NF-kappaB ligand (RANKL)-induced osteoclast differentiation and bone resorption: mice deficient in IFN signaling exhibit decreased bone mass accompanied by the activation of osteoclastogenesis. On the other hand, an unexpected increase in bone mass was observed in Stat1-deficient mice, indicating that Stat1 has a hitherto unknown function in the regulation of bone formation. Indeed, Stat1 was found to have a unique, non-canonical function as a cytoplasmic attenuator of Runx2, a key transcription factor for osteoblast differentiation. Thus, the loss of Stat1 results in excessive activation of Runx2 and osteoblast differentiation, thereby tipping the balance in favor of bone formation over bone resorption. This is an interesting example in which a latent transcription factor attenuates the activity of another transcription factor in the cytoplasm, and reveals a novel regulatory mechanism of bone remodeling by immunomodulatory molecules. Here, we summarize recent advances in the study of Stat1 and IFNs in the context of osteoimmunology, including latest reports that question whether the inhibitory function of Stat1 in chondrocytes is responsible for dwarfism in achondroplasia. PMID- 15546141 TI - An investigation into the effectiveness of bibliotherapy and minimal contact interventions in the treatment of panic attacks. AB - The present study investigated the effectiveness of bibliotherapy and minimal therapist-contact interventions in the treatment of panic attacks. Individuals were randomly assigned to one of three conditions: (1) bibliotherapy alone (BT); (2) bibliotherapy plus phone contact (BT+PC); or (3) phone contact alone (PC). Assessment (pre- and post-treatment) and treatment (8 weeks in duration) were conducted via mail and phone. Individuals receiving BT and BT+PC exhibited significant reductions from pre- to post-treatment on panic cognitions and fear of having a panic attack. Individuals receiving BT+PC exhibited significant reductions from pre- to post-treatment on panic symptoms and avoidance. In addition, individuals in the BT and BT+PC groups were more likely to exhibit clinically significant improvement on most dependent measures relative to PC alone. On some measures, individuals in the BT+PC group did clinically better than individuals in the BT group. Results of the present study also suggest that diagnosis may play some role in outcome. PMID- 15546142 TI - Trafficking and localization of platinum complexes in cisplatin-resistant cell lines monitored by fluorescence-labeled platinum. AB - Cisplatin is a chemotherapeutic agent commonly used in the treatment of a wide variety of malignant tumors. Resistance to cisplatin represents a major obstacle to effective cancer therapy because clinically significant levels of resistance quickly emerge after treatment. Based on previous studies indicating abnormal plasma membrane protein trafficking in cisplatin-resistant (CP-r) cells, Fluorescence (Alexa Fluor)-labeled cisplatin was used to determine whether this defect altered the trafficking and localization of cisplatin by comparing drug sensitive KB-3-1 and KB-CP-r cells. Alexa Fluor-cisplatin was readily internalized and localized throughout the KB-3-1 cells, but overall fluorescence decreased in KB-CP-r cells, as detected by flow cytometry (FACS) and confocal microscopy. Only punctate cytoplasmic staining was observed in KB-CP-r cells with less fluorescence observed in the nucleus. Colocalization experiments with a Golgi-selective stain indicate the involvement of Golgi-like vesicles in initial intracellular processing of Alexa Fluor conjugated cisplatin complexes. As detected using an antibody to Alexa Fluor-cisplatin, cisplatin complex-binding proteins (CCBPs) were reduced in membrane fractions of single-step cisplatin resistant KB-CP.5 cells, and increased in the cytoplasm of KB-CP.5 cells compared to KB-3-1 cells. CCBPs localized to lower density fractions in KB-CP.5 cells than in KB-3-1 cells as determined by iodixanol gradient centrifugation. In summary, inappropriate trafficking of CCBPs might explain resistance to cisplatin in cultured cancer cells, presumably because membrane binding proteins for cisplatin are not properly located on the cell surface in these cells, but are instead trapped in low density vesicles within the cytoplasm. PMID- 15546143 TI - Psychological mindedness and awareness of self and others. AB - The major goal of this study was to explore the relationship among psychological mindedness (PM) and several facets of awareness, including a general sense of mindfulness (Mindful Attention Awareness Scale; Brown & Ryan, 2003), as well as more specific awareness of self (self-consciousness scale; Fenigstein, Scheier, & Buss, 1975) and others (Interpersonal Reactivity Index; Davis, 1980). Participants were 103 undergraduate students at an urban liberal arts college. Results indicated that PM (PM Scale; Conte, Plutchik, Jung, Picard, Karasu, & Lotterman, 1990) is related to mindfulness (r = .41, p < .01), private self consciousness (r = .27, p < .05), as well as cognitive (r = .30, p < .01) and affective (r = .35, p < .01) indices of empathy. Self-consciousness and empathy explained a significant amount of variance in PM in a simultaneous-entry multiple regression. These findings support theoretical claims that PM involves awareness of self and others. PMID- 15546144 TI - Cognitive-behavioral therapy for PTSD in the real world: do interpersonal relationships make a real difference? AB - The goal of this effectiveness study was to investigate the role of pre-treatment interpersonal relationship functioning in two forms of group cognitive-behavioral treatment (CBT) for veterans with PTSD. Analysis of data from 45 veterans who completed either trauma- or skills-focused CBT indicated no overall differences between the two treatments in PTSD symptomatology, alcohol abuse, or violence perpetration at four months post-treatment. However, there was a stronger inverse relationship between intimate relationship functioning and violence outcomes in the trauma-focused group versus the skills-focused group. While no differences in violence outcomes were found between the treatments at poorer levels of pre treatment intimate relationship functioning, those receiving trauma-focused treatment with better pre-treatment intimate relationships reported less violence. Extended relationship functioning and violence outcomes were less strongly associated in the trauma-focused group versus the skills-focused group. The theoretical implications of these results, as well as the clinical opportunities to improve CBT for PTSD by capitalizing on patients' relationships, are discussed. PMID- 15546145 TI - Origins of bidirectional replication of Epstein-Barr virus: models for understanding mammalian origins of DNA synthesis. AB - Epstein-Barr virus (EBV), provides unique advantages to understand origins of replication in higher eukaryotes. EBV establishes itself efficiently in infected B lymphocytes, where it exists as a 165 kb, circular chromosome which is duplicated once per cell cycle (Adams [1987] J Virol 61:1743-1746). Five to twenty copies of the EBV chromosome are usually present in each cell, increasing the signal/noise ratio for mapping and analyzing its replication origins. Remarkably only one viral protein is required for the synthesis and partitioning of the viral chromosomes: EBV nuclear antigen-1, or EBNA1. EBV uses distinct origins related to the ARS1 origin of Saccharomyces cerevisiae and to that of the dihydrofolate reductase (DHFR) locus in Chinese hamster ovary (CHO) cells [Bogan et al., 2000]. We shall review the properties and the regulation of these two kinds of origins in EBV and relate them to their cellular cousins. PMID- 15546146 TI - Maintaining psychology's scientific and professional credibility and ethical responsibility to self-regulate: a comment on "validations" of mental health assessment instruments. AB - Recent growth of "managed" mental health care in the United States has spawned huge demand for products that draw on one of psychology's most well developed subdisciplines, tests and measurement. The commercial potential of mental health assessment instruments intended for widespread use, to meet what Kraus, Seligman, and Jordan (this volume) describe as "an industry-wide surge in outcome evaluations in naturalistic ... settings," necessarily raises conflict of interest dilemmas for those who develop and market them. The American Psychological Association has devoted intensive effort to the preceding issue as it pertains to other aspects of the science and practice of clinical psychology. Comparable attention has not been focused recently on the development and marketing of assessment instruments. This Comment highlights the issue and suggests types of self-regulatory actions that might be taken, e.g., requiring and publishing full disclosure statements of authors' relationships to companies that market instruments like the Treatment Outcome Package in psychometric articles in which they are evaluated. PMID- 15546147 TI - Validation of a behavioral health treatment outcome and assessment tool designed for naturalistic settings: The Treatment Outcome Package. AB - In 1994, the American Psychological Association and the Society for Psychotherapy Research convened a Core Battery Conference to develop a set of criteria for the selection of a universal core battery that could be used as a common outcome tool across all outcome studies. The Treatment Outcome Package (TOP) is a behavioral health assessment and outcome battery with modules for assessing a wide array of behavioral health symptoms and functioning, demographics, case-mix, and treatment satisfaction. It was developed to follow the design specifications set forth by the Core Battery Conference, but also to ensure the battery's applicability to naturalistic treatment settings in which randomization may be impossible. In this article we discuss a number of studies that evaluate the initial psychometrics of the items that comprise the mental health symptom and functional modules of the TOP. We conclude that the TOP has an excellent factor structure, good test-retest reliability, promising initial convergent and discriminant validity, measures the full range of pathology on each scale, and has some ability to distinguish between behavioral health clients and members of the general population. PMID- 15546148 TI - In search of the molecular mechanism by which small stress proteins counteract apoptosis during cellular differentiation. AB - Many differentiation programs are accompanied by an increase in small heat shock proteins (sHsps) level. Most of the time transient, this accumulation takes place during the early phase of the process and is correlated with the growth arrest that precedes the differentiation. Important biochemical modifications of sHsps occur, such as changes in phosphorylation and oligomerization. The fact that these proteins are induced independently of the signal that triggers differentiation, of the differentiation type, and of the cell type strongly suggests their involvement in fundamental mechanisms of cellular differentiation. Moreover, impairment of sHsps accumulation leads to abortion of the differentiation program and, subsequently, to a massive commitment to cell death. Recent advances in this field of research are presented as well as the hypothesis that should be tested to unravel the mode of action of these proteins during cellular differentiation. PMID- 15546149 TI - Lipopolysaccharide affects Golli expression and promotes proliferation of oligodendrocyte progenitors. AB - Proliferation of oligodendrocyte progenitor cells (OPCs) is important for initial myelination as well as for remyelination in demyelinating diseases. Previously, we showed that numerous OPCs and activated microglia, are present around multiple sclerosis lesions, and that they accumulate Golli proteins. Golli proteins, present in both neuronal and immune cells, might have a role in the immune processes, as well as in development of neurons and oligodendrocytes. We hypothesize that Golli proteins, generated by microglia in response to inflammation, promote proliferation of OPCs. To test this hypothesis, we induced inflammation in neonatal mouse brain slice culture with bacterial endotoxin lipopolysaccharide (LPS). Treated slices showed an increase in the number of OPCs. Several results support the notion that this effect of LPS is conveyed through activation of microglia and upregulation of Golli proteins. First, LPS treated brain slices have increased expression of Golli proteins observed by immunofluorescence and Western blot analysis. Second, Golli proteins were demonstrated only in the conditioned medium from LPS-treated microglial cell cultures (LPS-MCM), and were absent in either the conditioned media from LPS treated astrocytes or the control media. Third, proliferation of purified OPCs was promoted with LPS-MCM or Golli proteins, but not with LPS alone. Taken together, these results demonstrate that microglia and/or microglia secreted factors, are necessary for the LPS-promoted proliferation of OPCs and suggest possible involvement of Golli proteins as one of mediators in this process. PMID- 15546150 TI - Mammalian cell delivery via aerosol deposition. AB - The objective of this study was to test the hypothesis that bovine dermal fibroblasts can survive aerosol delivery via an airbrush with mean cell survival rates greater than 50%. This technology has great implications for burn and other wound therapies, for delivery of genetically altered cells in gene therapies, and for tissue engineering with tissue scaffolds. Bovine dermal fibroblasts were suspended at a concentration of 200,000 cells/mL in Hank's Balanced Salt Solution, and delivered into six-well tissue culture plates using a Badger 100G airbrush. Cells were delivered through three nozzle diameters (312, 484, and 746 microm) at five different air pressures (41, 55, 69, 96, and 124 kPa). Nine repetitions were performed for each treatment group, and cell viability was measured using trypan blue exclusion assay. Mean cell viability ranged from 37 to 94%, and depended on the combination of nozzle diameter and delivery pressure (p < 0.0001). Linear regression analysis was used to develop a stochastic model of cell delivery viability as a function of nozzle diameter and delivery air pressure. This study demonstrates the feasibility of using an airbrush to deliver viable cells in an aerosol to a substrate. PMID- 15546151 TI - New Huntington disease mutation arising from a paternal CAG34 allele showing somatic length variation in serially passaged lymphoblasts. AB - The analysis of somatic CAG triplet variation in lymphoblastoid cell lines from subjects carrying alleles of intermediate length (IA(33CAG) and IA(34CAG)) in Huntington disease (HD) gene disclosed instability in the DNA of the person, from whom a new expansion mutation of 45 triplets originated. The triplet size increased after about 30 passages in cell cultures in lymphoblasts with the IA(34) genotype. Lymphoblasts may provide an appropriate model for studying repeat instability in subjects with poly(CAG) repeat disorders. HD shows somatic, in addition to germ-line instability, highlighting the propensity to somatic CAG variation in human cells even with repeat numbers under the expanded edge. Factors potentially cis acting with the mutation, other than those reported in this study (CCG polymorphic stretch, the deletion of the glutamic acid residue at position 2642 and the 4-codon segment between CAG and CCG polymorphisms), should be searched for and analyzed. PMID- 15546152 TI - Humoral and contact interactions in astroglia/stem cell co-cultures in the course of glia-induced neurogenesis. AB - Astroglial cells support or restrict the migration and differentiation of neural stem cells depending on the developmental stage of the progenitors and the physiological state of the astrocytes. In the present study, we show that astroglial cells instruct noncommitted, immortalized neuroectodermal stem cells to adopt a neuronal fate, while they fail to induce neuronal differentiation of embryonic stem cells under similar culture conditions. Astrocytes induce neuron formation by neuroectodermal progenitors both through direct cell-to-cell contacts and via short-range acting humoral factors. Neuron formation takes place inside compact stem cell assemblies formed 30- 60 h after the onset of glial induction. Statistical analyses of time-lapse microscopic recordings show that direct contacts with astrocytes hinder the migration of neuroectodermal progenitors, while astroglia-derived humoral factors increase their motility. In non-contact co-cultures with astrocytes, altered adhesiveness prevents the separation of frequently colliding neural stem cells. By contrast, in contact co cultures with astrocytes, the restricted migration on glial surfaces keeps the cell progenies together, resulting in the formation of clonally proliferating stem cell aggregates. The data indicate that in vitro maintained parenchymal astrocytes (1) secrete factors, which initiate neuronal differentiation of neuroectodermal stem cells; and (2) provide a cellular microenvironment where stem cell/stem cell interactions can develop and the sorting out of the future neurons can proceed. In contrast to noncommitted progenitors, postmitotic neuronal precursors leave the stem cell clusters, indicating that astroglial cells selectively support the migration of maturing neurons as well as the elongation of neurites. PMID- 15546153 TI - Mechanistic and epidemiologic considerations in the evaluation of adverse birth outcomes following gestational exposure to statins. AB - The cholesterol-lowering "statin" drugs are contraindicated in pregnancy, but few data exist on their safety in human gestation. We reviewed case reports for patterns suggesting drug-related effects on prenatal development and considered a variety of mechanisms by which such effects, if confirmed, might occur. This uncontrolled case series included all FDA reports of statin exposures during gestation, as well as others from the literature and from manufacturers. Exposures and outcomes were reviewed and were tabulated by individual drug. Age specific rates of exposure to each drug among women of child-bearing age were estimated. Of 214 ascertained pregnancy exposures, 70 evaluable reports remained after excluding uninformative cases. Among 31 adverse outcomes were 22 cases with structural defects, 4 cases of intrauterine growth restriction, and 5 cases of fetal demise. There were two principal categories of recurrent structural defects: cerivastatin and lovastatin were associated with four reports of severe midline CNS defects; simvastatin, lovastatin, and atorvastatin were all associated with reports of limb deficiencies, including two similar complex lower limb defects reported following simvastatin exposure. There were also two cases of VACTERL association among the limb deficiency cases. All adverse outcomes were reported following exposure to cerivastatin, simvastatin, lovastatin, or atorvastatin, which are lipophilic and equilibrate between maternal and embryonic compartments. None were reported following exposure to pravastatin, which is minimally present in the embryo. Statins reaching the embryo may down-regulate biosynthesis of cholesterol as well as many important metabolic intermediates, and may have secondary effects on sterol-dependent morphogens such as Sonic Hedgehog. The reported cases display patterns consistent with dysfunction of cholesterol biosynthesis and Sonic Hedgehog activity. Controlled studies are needed to investigate the teratogenicity of individual drugs in this class. PMID- 15546154 TI - Blue light differentially alters cellular redox properties. AB - Blue light (lambda = 380-500 nm) historically has been used to initiate polymerization of biomaterials and recently has been proposed as a therapeutic agent. New evidence suggests that cell-type-specific responses result from redox changes induced by exposure to blue light. Cultured cells were exposed to defined doses of blue light, equivalent to exposure times of 10 s and 2 min, to achieve energies of 5 J/cm2 and 60 J/cm2, respectively, after which (a) viable cell number, (b) cellular protein profiles, (c) mitochondrial succinate dehydrogenase (SDH) activity, (d) total reactive oxygen species (ROS), and (e) induction of apoptosis were compared to that of nonexposed control cultures. Results showed that blue-light exposure arrested monocyte cell growth and increased levels of peroxiredoxins. SDH activity of normal epidermal keratinocytes (NHEK) was slightly enhanced by blue light, whereas identical treatment of OSC2 oral tumor cells resulted in significant suppression of SDH activity. Blue-light exposure generally induced higher levels of total ROS in OSC2 cells than in NHEK. Finally, only OSC2 cells exhibited signs of apoptosis via Annexin V staining following exposure to blue light. These data support the central hypothesis that blue light induces an oxidative stress response in cultured cells resulting in cell-type specific survival outcomes. The identification of oxidative stress as a mediator of the effects of blue light is a critical first step in defining its biological risks and therapeutic opportunities. PMID- 15546155 TI - Neuroglial activation and neuroinflammation in the brain of patients with autism. AB - Autism is a neurodevelopmental disorder characterized by impaired communication and social interaction and may be accompanied by mental retardation and epilepsy. Its cause remains unknown, despite evidence that genetic, environmental, and immunological factors may play a role in its pathogenesis. To investigate whether immune-mediated mechanisms are involved in the pathogenesis of autism, we used immunocytochemistry, cytokine protein arrays, and enzyme-linked immunosorbent assays to study brain tissues and cerebrospinal fluid (CSF) from autistic patients and determined the magnitude of neuroglial and inflammatory reactions and their cytokine expression profiles. Brain tissues from cerebellum, midfrontal, and cingulate gyrus obtained at autopsy from 11 patients with autism were used for morphological studies. Fresh-frozen tissues available from seven patients and CSF from six living autistic patients were used for cytokine protein profiling. We demonstrate an active neuroinflammatory process in the cerebral cortex, white matter, and notably in cerebellum of autistic patients. Immunocytochemical studies showed marked activation of microglia and astroglia, and cytokine profiling indicated that macrophage chemoattractant protein (MCP)-1 and tumor growth factor-beta1, derived from neuroglia, were the most prevalent cytokines in brain tissues. CSF showed a unique proinflammatory profile of cytokines, including a marked increase in MCP-1. Our findings indicate that innate neuroimmune reactions play a pathogenic role in an undefined proportion of autistic patients, suggesting that future therapies might involve modifying neuroglial responses in the brain. PMID- 15546156 TI - A convergent route to enantiomerically pure higher [n-2]triangulanedimethanol derivatives and [n]triangulanes (n>/=7). PMID- 15546157 TI - Disregarded use of herbal medical products and dietary supplements among surgical and medical patients as estimated by home inspection and interview. AB - PURPOSE: More and more patients use herbal medical products (HMP) and dietary supplements (DS). Due to the possibility of drug interactions and side effects, it is important that physicians are aware of the use. The aim of the present cross sectional survey was to analyse the consumption of HMP and DS among patients recently discharged from two hospital departments. METHODS: Patients were visited within 1 week after discharge and interviewed about their use of HMP and DS. Stored products were inspected and registered. Hospital files and discharge letters were examined to establish the frequency of registration. RESULTS: Totally, 83 surgical and 117 medical patients were included (n = 200), 139 patients (70%) were women. 53 patients (27%) stored no HMP or DS, whereas the home inventories of 147 patients (74%) comprised 343 products. 116 patients (58%) used HMP or DS daily and 25 patients (13%) used the products on demand. The most frequently used product was multivitamins (82 patients = 41%). Totally, 61% of the products were used on the patients' own initiative. In 3% of the used products, patients were aware of possible side effects. No patients were aware of possible drug interactions. Only 21% of the 211 HMP and DS used daily prior to admission were recorded in the hospital files. CONCLUSIONS: The use of HMP and DS was frequent among surgical and medical patients. The use was often not recorded in the hospital files and patients' knowledge of possible side effects and drug interactions was minimal. PMID- 15546158 TI - Over-expression of TGF-beta1 in Smad4-deficient human oral carcinoma cells causes tumour regression in vivo by mechanisms that sensitize cells to apoptosis. AB - We have shown previously that transforming growth factor-beta (TGF-beta) is a potent tumour suppressor in Smad4-deficient human malignant oral keratinocytes but the mechanism by which this occurs is unknown. In the present study, we show that over-expression of TGF-beta1 causes regression of tumours derived from Smad4 deficient oral keratinocytes transplanted orthotopically to athymic mice. Further, tumour regression is associated with the induction of apoptosis without changes in cell proliferation. In vitro, TGF-beta1 did not induce apoptosis directly in these cells but sensitized cells to cisplatin, but not Fas, -induced cell death. The data suggest that TGF-beta1 induces tumour regression in vivo by Smad4-independent pathways that sensitize keratinocytes to mitochondrial-mediated apoptosis. PMID- 15546159 TI - Impact of regulatory labeling for troglitazone and rosiglitazone on hepatic enzyme monitoring compliance: findings from the state of Ohio medicaid program. AB - PURPOSE: Troglitazone, the first drug of the thiazolidinediones class for type II diabetes, was first marketed in March 1997 and was removed from the U.S. market 36 months later after 90 cases of liver failure were reported despite multiple warnings containing liver enzyme monitoring recommendations. Rosiglitazone has been available since June 1999 and is still on the market. The purpose of this study was to evaluate the impact of labeled hepatic enzyme monitoring for troglitazone and rosiglitazone. METHODS: Drug cohorts were assembled, using population-based fee-for-service Medicaid claims, for patients between 18 and 65 years of age who had received at least one troglitazone (n = 7226) or rosiglitazone (n = 1480) prescription between 1 April, 1997, and 21 March, 2000. The outcome of interest was the percentage of patients, based on their first treatment episode, who had baseline and post-baseline liver enzyme testing. RESULTS: Overall baseline testing was under 9% before regulatory actions, increased to 14% after the first two 'Dear Doctor' letters issued by the FDA in October and December 1997, and peaked to about 26% afterwards. Coincident with the marketing of rosiglitazone and the fourth 'Dear Doctor' letter issued in June 1999, baseline testing dropped to 18%. Baseline testing increased 2.5-fold (race sex-age adjusted) after regulatory action. Achieving 50% post-baseline testing took approximately 6 months for both drugs. CONCLUSION: Regulatory actions had only modest effects on the incidence of liver monitoring. More effective and timely communication strategies, health provider prescribing interventions and modification of health provider behaviors to enhance compliance with recommended risk management measures need to be identified, evaluated and implemented. PMID- 15546160 TI - Bone marrow cells engraft within the epidermis and proliferate in vivo with no evidence of cell fusion. AB - In adults, bone marrow-derived cells (BMDC) can contribute to the structure of various non-haematopoietic tissues, including skin. However, the physiological importance of these cells is unclear. This study establishes that bone marrow derived epidermal cells are proliferative and, moreover, demonstrates for the first time that BMDC can localize to a known stem cell niche: the CD34-positive bulge region of mouse hair follicles. In addition, engraftment of bone marrow cells into the epidermis is significantly increased in wounded skin, bone marrow derived keratinocytes can form colonies in the regenerating epidermis in vivo, and the colony-forming capacity of these cells can be recapitulated in vitro. In some tissues this apparent plasticity is attributed to differentiation, and in others to cell fusion. Evidence is also provided that bone marrow cells form epidermal keratinocytes without undergoing cell fusion. These data suggest a functional role for bone marrow cells in epidermal regeneration, entering known epidermal stem cell niches without heterokaryon formation. PMID- 15546161 TI - Surgeon's recommendation, perceived operative efficacy and age dictate treatment choice by Chinese women facing breast cancer surgery. AB - PURPOSE: to identify factors influencing Chinese women's choices between breast conserving therapy (BCT), mastectomy (MRM) or MRM followed by breast reconstruction (MRM+R). METHODS: of 405/443 Hong Kong Chinese women receiving surgery for early breast cancer who were interviewed one week post-surgery about their pre-surgical consultation, available treatment alternatives, whether their surgeons had indicated a surgical preference, perceived efficacy of the surgical options and considerations influencing their treatment decisions (TDM), 198 (49%) reported they were offered a choice of surgery. RESULTS: among women offered a choice of surgery, BCT was chosen by 34/43 (79%) of women whose surgeons recommended BCT but by only 34/96 (37%) of women whose surgeons expressed no treatment recommendation. Multivariate adjustment showed women choosing MRM were influenced more by avoiding both cancer recurrence (p = 0.003) and further treatment (p = 0.009) when choosing surgical option than women choosing BCT. In contrast, women choosing MRM+R and BCT, placed more emphasis on appearance (p < 0.001) and body image (p < 0.001) concerns as influencing treatment choice than did women who chose MRM. CONCLUSION: survival concerns rather than physical appearance, age and lack of recommendation push Chinese women to choose MRM as BCT is, incorrectly often seen as less efficacious. Recommending BCT increases BCT choice. PMID- 15546162 TI - pDUAL, a multipurpose, multicopy vector capable of chromosomal integration in fission yeast. AB - A novel series of plasmid vectors named pDUAL have been developed. These vectors enable one to introduce not only multicopies of genes with episomal maintenance but also a single copy with chromosomal integration into the fission yeast, Schizosaccharomyces pombe. The multicopy plasmids can be easily converted to fragments for chromosomal integration by digestion of the plasmids with a certain restriction endonuclease before transformation of the yeast cells. The resultant fragments, lacking the autonomously replicating sequence, are designed for targeting into the chromosomal leu1 locus by homologous recombination. Whether the transformants are the results of episomal maintenance of the plasmid or homologous gene targeting can be readily checked by their requirement for uracil or leucine, or by the PCR diagnostic analysis. Furthermore, we propose the use of pDUAL derivatives for PCR-based chromosomal tagging of a gene to introduce several tags into 5'-terminus of a gene, employing a set of primers. Using these all-in-one vectors, a suitable mode of expression of a cloned gene can be selected for individual analysis without any complicated subcloning processes. PMID- 15546164 TI - Research Corporation and John Schaefer. PMID- 15546163 TI - Suppression of distant pulmonary metastasis of MDA-MB 435 human breast carcinoma established in mammary fat pads of nude mice by retroviral-mediated TIMP-2 gene transfer. AB - BACKGROUND: Previous studies have shown that TIMP-2 overexpression is a useful therapeutic tool for inhibiting tumor growth and invasion in animals. However, it has not been reported whether genetic manipulation for TIMP-2 overexpression can induce an inhibitory effect on spontaneous metastasis from the primary tumor site to other organs such as lungs or lymph nodes in an animal model. METHODS: The present studies describe the effects of retrovirus-mediated TIMP-2 gene transfer into human breast cancer cell lines on the in vitro invasion of the tumor cells or the in vivo growth in nude mouse. Here we also used retroviral-mediated TIMP-2 overexpression by intratumoral injection for suppression of metastasis in human breast carcinoma established in the mammary fat pad of nude mice. RESULTS: As expected, overexpression of TIMP-2 inhibited matrix metalloprotenase (MMP) activity and invasion of the tumor cells. Also, the growth rate of tumors grafted with the breast cancer cells transduced with the retrovirus vector encoding TIMP 2 cDNA was significantly slower than the growth rate of tumors grafted with the breast cancer cells transduced with a control retrovirus vector. Furthermore, single intratumoral injection of the TIMP-2 retrovirus-producing cells into human breast tumor tissue established in mammary fat pads of nude mice showed a dramatic decrease in size and number of lung metastatic tumors. CONCLUSIONS: Retrovirus-mediated TIMP-2 gene transfer into human breast cancer cells is able to down-regulate invasion and show that tumor-derived angiogenesis is reduced. In this model, retroviral-mediated transduction of TIMP-2 cDNA into a limited population of human tumor cells inhibits tumor growth and prevents distant pulmonary metastasis. These results indicate that it may not be necessary to deliver and express these genes in every single tumor cell as long as the level of expression in a limited number of transduced cells is sufficient to prevent the excessive breakdown of the extracellular matrix. PMID- 15546165 TI - Nursing 2004 salary survey. PMID- 15546166 TI - Putting the brakes on necrotizing fasciitis. PMID- 15546167 TI - A "current" choice for hemodynamic monitoring. PMID- 15546168 TI - Preventing CABG donor site infection. PMID- 15546169 TI - The nurse poachers. PMID- 15546170 TI - Central Europe dabbles in private health. PMID- 15546171 TI - Mastalgia: an adverse effect of sildenafil. PMID- 15546172 TI - Borst (Jadassohn): a misnomer. PMID- 15546173 TI - Is routine early invasive management of non-ST-segment elevation myocardial infarction beneficial in elderly patients? PMID- 15546174 TI - Incidental metastases of well-differentiated thyroid carcinoma in lymph nodes of patients with squamous cell head and neck cancer: eight cases with a review of the literature. AB - The examination of a large series of cervical lymph nodes in patients with head and neck cancer revealed the presence of incidental metastases of occult thyroid carcinoma in eight patients, of which six cases were squamous cell carcinoma of glottic and supraglottic sites of the larynx and two cases were pyriform sinus and tongue carcinomas. Three patients had two lymph nodes and the remaining patients had one lymph node each involved. The nodal chains affected were the jugular (n=5; level IV), Kuttner (level II), supraomohyoid (level III) and supraclavicular (level VI). In four cases, a subtotal thyroidectomy or unilateral lobectomy was performed during laryngectomy (for surgical reasons) or after histologic nodal examination; a minimal focus of thyroid papillary carcinoma was detected in one patient. Three of eight patients died from recurrence of the squamous cell carcinoma; no case presented clinical evidence of thyroid malignancy. The differential diagnosis from benign thyroid heterotopia was based on the presence of minimal nuclear atypia. The choice of treatment of patients with a coexisting neoplasm characterized by poor prognosis is difficult, and contrasting opinions exist regarding the use of radical thyroidectomy and the subsequent management. As reported in the literature (66 cases), the more aggressive squamous cell carcinoma will determine the prognosis of these patients; in fact, only one of the referred cases died of cerebellar metastases of the thyroid cancer. Our results emphasize the importance of an accurate re evaluation and follow-up of patients with incidental occult metastases for detection of a primary thyroid tumor. In the general population, this incidental nodal involvement may be related to a minimal occult thyroid carcinoma. PMID- 15546175 TI - Steroid injection to vocal nodules using fiberoptic laryngeal surgery under topical anesthesia. AB - Since 1990, we have performed steroid injection into the vocal fold by fiberoptic laryngeal surgery (FLS) under local anesthesia. In this study, the usefulness of this method was evaluated in 28 patients with vocal nodules. Under monitoring using a fiberoptic laryngoscope, a curved injection needle was inserted via the oral cavity and steroid was injected. Endoscopic findings showed that the vocal nodule had disappeared in 17 patients of the 27 patients and decreased in 10 after injection. The maximum phonation time was 10.9 s before operation and 13.9 s after operation, showing a significant increase (P<0.05), and the mean flow rate also showed a significant improvement (P<0.05). The patients self-rating concerning hoarseness demonstrated great improvement after injection. This technique can be performed under local anesthesia in combination with voice therapy on an outpatient basis, and it is considered to be useful for treating vocal nodules. PMID- 15546176 TI - Preoperative imaging of chronic sinusitis by multislice computed tomography. AB - The aim of this study was to evaluate whether multislice CT enables quality improvement and dose reduction in the imaging of the paranasal sinuses, especially when using secondary reconstructions. We compared the imaging quality of direct CT scans and secondary reconstructions of single-slice CT (SSCT) as a criterion standard with multislice CT (MSCT) of the paranasal sinuses in 80 patients suspected of having chronic sinusitis. Coronary secondary reconstructions were calculated from all transversal CT data sets. Coronary reconstructions of transversal MSCT showed a significantly better image quality compared with coronary reconstructions of SSCT. Because of the absence of dental metal artifacts, coronary reconstructions of MSCT were superior even to direct coronary images of SSCT. MSCT offered a superior examination quality compared to SSCT. A halving of radiation dosage can be reached by eliminating one examination plane. PMID- 15546177 TI - Identification of the gonad-specific anion transporter SLCO6A1 as a cancer/testis (CT) antigen expressed in human lung cancer. AB - Serological analysis of recombinant cDNA expression libraries (SEREX) has led to the identification of many of the antigens recognized by the immune system of cancer patients, which are collectively referred to as the cancer immunome. We used SEREX to screen a testicular cDNA expression library with sera obtained from non-small cell lung cancer patients and isolated cDNA clones for 82 antigens. These included a total of 31 antigens previously identified by SEREX, and 51 that did not match entries in the Cancer Immunome Database and were considered newly identified antigens. Overall, the antigens comprised 62 known proteins and 20 uncharacterized gene products. Six antigens (NY-TLU-6, -37, -39, -57, -70, -75) were identified as putative cell surface proteins that are potential targets for monoclonal antibody-based immunotherapy. Of these, the gonad-specific anion transport protein SLCO6A1 (NY-TLU-57) was shown to be tissue-restricted. RT-PCR showed it to be expressed strongly only in normal testis, and weakly in spleen, brain, fetal brain, and placenta. In addition, NY-TLU-57 mRNA was found in lung tumor samples (5/10) and lung cancer cell lines (6/11), as well as bladder (5/12) and esophageal (5/12) tumor samples. These data suggest that SLCO6A1 is a putative cancer/testis (CT) cell surface antigen of potential utility as a target for antibody-based therapy for a variety of tumor types. The analysis also permits us to estimate the eventual size of the SEREX-defined cancer immunome at around 4000 genes. This emphasizes the importance of continued SEREX screening to define the cancer immunome. PMID- 15546179 TI - Synthesis of phosphate-branched oligonucleotides. AB - A solid-phase synthesis for phosphate-branched oligonucleotides is described. The method is based on coupling of a single nucleoside phosphorodiamidite to terminal hydroxyl functions of two solid-supported oligonucleotides. After oxidation of the phosphite triester obtained to a phosphate triester, the third branch is assembled by conventional phosphoramidite chemistry. PMID- 15546180 TI - Targeting malaria with polyamines. AB - During the asexual cycle of Plasmodium falciparum within the host erythrocyte, the parasite induces a stage-dependent elevation in the levels of polyamines by increased metabolism and uptake of extracellular pools. Polyamine amides of N methylanthranilic acid have been synthesized which have embedded within them putrescine, spermidine, or spermine and from a charge perspective mimic natural polyamines. The interaction of these polyamine conjugates with human erythrocytes infected with malaria is described using fluorescent microscopy. The fluorescent spermine mimic was the only probe to show measurable intracellular accumulation. This was observed in late stage development but not in the ring stages or in uninfected erythrocytes. PMID- 15546181 TI - Cytosolic delivery of macromolecules. 3. Synthesis and characterization of acid sensitive bis-detergents. AB - A serious limitation that precludes utilization of single-tailed, pH-sensitive detergents for the cytosolic delivery of macromolecules is their low limit of incorporation in stable liposomal formulations. To address this issue, we have prepared two Gemini surfactants or 'bis-detergents' by cross-linking the headgroups of single-tailed, tertiary amine detergents through oxyethylene (BD1) or acid-labile acetal (BD2) moieties. The membrane-bound pK(a) of the twin tertiary amine headgroups was determined to be 6.37 +/- 0.36 using a fluorescence based assay. As evidenced by thin-layer chromatography, BD2was hydrolyzed under acidic conditions (pH 5.0) with an approximate half-life of 3 h at 37 degrees C, while BD1 remained stable. Low pH-induced collapse of liposomes containing acid labile BD2 into micelles was more facile than that of BD1. With BD1, the process appeared to be reversible in that aggregation of micelles was observed at basic pH. The irreversible lamellar-to-micellar transition observed with BD2-containing liposomes can possibly be attributed to acid-catalyzed hydrolysis of the acetal cross-linker, which generates two detergent monomers within the bilayer. Liposomes composed of 75 mol % bis-detergent and 25 mol % phosphatidylcholine were readily prepared and could entrap macromolecules such as polyanionic dextran of MW 40 kDa with moderate efficiency. The ability of BD2-containing liposomes to promote efficient cytosolic delivery of antisense oligonucleotides was confirmed by (a) their diffuse intracellular distribution seen in fluorescence micrographs, and (b) the up-regulation of luciferase in an antisense functional assay. The low pH-responsive, bis-detergent constructs described herein are suitable for triggered release strategies targeted to acidic intracellular or interstitial environments. PMID- 15546182 TI - Synthesis and in vitro testing of J591 antibody-dendrimer conjugates for targeted prostate cancer therapy. AB - Targeted therapeutics using antibodies are an attractive option over conventional cancer chemotherapeutics due to their potential to deliver a therapeutic specifically to cancer tissue without damaging normal tissue. However, there are known problems with immunoconjugates such as decreased immunoreactivity and poor solubility. Using dendrimers as carriers for these agents has the potential to resolve these issues. We synthesized J591 anti-PSMA (prostate specific membrane antigen) antibody dendrimer conjugates containing fluorophores on the dendrimer. The in vitro studies of these conjugates show that they specifically bind to cells expressing PSMA. Confocal microscopy experiments document the binding and internalization of these conjugates. This research encourages the further study of antibody-dendrimer-drug conjugates for use in targeted therapeutics. PMID- 15546183 TI - Synthesis and evaluation of a triplex-forming oligonucleotide pyrrolobenzodiazepine conjugate. AB - In most cases, unmodified oligonucleotides designed as antigene molecules are incapable of binding to DNA with sufficient stability to prevent gene expression. To stabilize binding to a polypurine tract in the HER-2/neu promoter, a triplex forming oligonucleotide (TFO) was conjugated to a pyrrolo[1,4]benzodiazepine (PBD), desmethyltomaymycin, and site-specific DNA binding was evaluated. An activated ester of the PBD moiety was conjugated by an acylation reaction to a free primary amine on a 50-atom aliphatic linker at the 5' end of the TFO. This long aliphatic linker was designed to provide a bridge from the major groove binding site of the TFO to the minor groove binding site of the PBD. Triplex formation by the resulting TFO-PBD conjugate occurred more slowly and with a nearly 30-fold lower affinity compared to an unconjugated TFO. PBD binding to the triplex target was demonstrated by protection from restriction enzyme digestion, and covalent binding to the exocyclic amino group of guanine was inferred by substituting specific guanines with inosines. Although the binding of the TFO was less efficient, this report demonstrates that in principle, TFOs can be used to direct the binding of a PBD to specific location. Further optimization of TFO-PBD conjugate design, likely involving optimization of the linker and perhaps placing a PBD at both ends of the TFO, will be needed to make gene modification robust. PMID- 15546184 TI - Influence of pendant arms bearing ligating groups on the structure of bismuth porphyrins: implications for labeling immunoglobulins used in medical applications. AB - The synthesis of two picket bismuth(III) porphyrins is reported, and their crystal structures are compared. The influence of the nature of the pickets, as well as their number, is discussed in terms of stability, kinetics of metalation, structure, and distortion of the porphyrin. Unexpectedly, the results indicate that the coordination sphere of bismuth is not affected by different types of distortion nor is the stability of the complex. For both types of complexes, the X-ray crystallography revealed that a single arm is actually in direct interaction with the bismuth cation, thereby allowing further functionalization on the porphyrin core as the linkage with immunoglobulins or anti-rabbit antibodies (both noted IgG), which have been used in diagnostic or therapeutic applications. PMID- 15546185 TI - Synthetic biocompatible cyclodextrin-based constructs for local gene delivery to improve cutaneous wound healing. AB - The localized, sustained delivery of growth factors for wound healing therapy is actively being explored by gene transfer to the wound site. Biocompatible matrices such as bovine collagen have demonstrated usefulness in sustaining gene therapy vectors that express growth factors in local sites for tissue repair. Here, new synthetic biocompatible materials are prepared and shown to deliver a protein to cultured cells via the use of an adenoviral delivery vector. The synthetic construct consists of a linear, beta-cyclodextrin-containing polymer and an adamantane-based cross-linking polymer. When the two polymers are combined, they create an extended network by the formation of inclusion complexes between the cyclodextrins and adamantanes. The properties of the network are altered by controlling the polymer molecular weights and the number of adamantanes on the cross-linking polymer, and these modifications and others such as replacement of the beta-cyclodextrin (host) and adamantane (guest) with other cyclodextrins (hosts such as alpha, gamma, and substituted members) and inclusion complex forming molecules (guests) provide the ability to rationally design network characteristics. Fibroblasts exposed to these synthetic constructs show proliferation rates and migration patterns similar to those obtained with collagen. Gene delivery (green fluorescent protein) to fibroblasts via the inclusion of adenoviral vectors in the synthetic construct is equivalent to levels observed with collagen. These in vitro results suggest that the synthetic constructs are suitable for in vivo tissue repair applications. PMID- 15546186 TI - Poly(3-guanidinopropyl methacrylate): a novel cationic polymer for gene delivery. AB - A cationic polymethacrylate with a guanidinium side group was designed in order to create a polymer with cell membrane-penetrating properties such as Tat or other arginine-rich peptides. The polymer, poly(3-guanidinopropyl methacrylate), abbreviated as pGuaMA, was synthesized by free radical polymerization. The DNA condensing properties of pGuaMA (Mw 180 kDa) were investigated via dynamic light scattering and zeta potential measurements, and small, positively charged particles (110 nm, +37 mV) were found. It was shown that polyplexes based on pGuaMA were able to transfect COS-7 cells efficiently in the absence of serum, while under the same conditions poly(arginine) (pArg) polyplexes did not show detectable transfection levels. Addition of a membrane-disrupting peptide, INF 7, derived from the influenza virus, to preformed pGuaMA polyplexes did result in approximately 2 times increased transfection levels. DLS, zeta potential measurements, gel electrophoresis, and ethidium bromide displacement measurements indicated that serum induced aggregation of the polyplexes at high polymer/plasmid ratios, while at low polymer/plasmid ratios the polarity of the polyplexes reversed likely due to adsorption of negatively charged proteins on their surface. Likely, the unfavorable interactions of pGuaMA polyplexes with serum proteins is the reason for the absent transfection activity of these polyplexes in the presence of serum. Confocal laser scanning microscopy indicated cellular internalization via endocytosis of both polyplexes and free polymer. Thus, pGuaMA polyplexes enter cells, as reported for other polyplexes, by endocytosis and not, as hypothesized, via direct membrane passage. PMID- 15546187 TI - Hydrotropic dendrimers of generations 4 and 5: synthesis, characterization, and hydrotropic solubilization of paclitaxel. AB - Polyglycerol dendrimers (PGDs) with 4-5 generations were synthesized and used to investigate the effect of dendritic architecture and its generation on aqueous solubilization of paclitaxel (PTX), a poorly water-soluble drug. Chemical and physical properties of the PGDs were characterized by NMR, MALDI-TOF mass, GPC, viscosity, and dynamic light scattering measurements. The PTX solubility in all the solutions of PGDs, even below 10 wt %, was much higher than that in PEG400 that is commonly used as a cosolvent or a hydrotropic agent. Increase in the PTX solubility by PGDs was dependent on the dendrimer generation. The dendritic structure was the reason for the enhanced solubility of PTX even at low concentrations. (1)H NMR spectra of PTX before and after mixing with PGDs in D(2)O suggested that the aromatic rings and some methyne groups of PTX were surrounded by PGDs. PGDs, which do not require hydrophobic segment as in polymeric micelles, provide an alternative method of hydrotropic solubilization of poorly soluble drugs. PMID- 15546188 TI - Tuning of intercalation and electron-transfer processes between DNA and acridinium derivatives through steric effects. AB - A series of acridinium derivatives 1-6, wherein steric factors have been varied systematically through substitution at the 9 position of the acridine ring, have been synthesized and their DNA interactions have been investigated by various biophysical techniques. The unsubstituted and methylacridinium derivatives 1 and 2 and the o-tolylacridinium derivative 6 exhibited high fluorescence quantum yields (Phi(f)() congruent with 1) and lifetimes (tau = 35, 34, and 25 ns, respectively), when compared with the arylacridinium derivatives 3-5. The acridinium derivatives 1 and 2 showed high DNA binding affinity (K = 7.3-7.7 x 10(5) M(-)(1)), when compared to the arylacridinium derivatives 3-5 (K = 6.9-10 x 10(4) M(-)(1)). DNA melting and viscosity studies establish that in the case of the aryl-substituted systems, the efficiency of DNA binding is in the order, phenyl > p-tolyl > m-tolyl >>>> o-tolyl derivative. The increase in steric crowding around the acridine ring hinders the DNA binding interactions and thereby leads to negligible binding as observed in the case of 6 (o-tolyl derivative). These results indicate that a subtle variation in the substitution pattern has a profound influence on the photophysical and DNA interactions. Further, they demonstrate that pi-stacking interactions of the ligands with DNA are essential for efficient electron transfer between the DNA bases and the ligands. These water soluble and highly fluorescent molecules which differ in their DNA binding mode can act as models to study various DNA-ligand interactions. PMID- 15546189 TI - Binding behavior of aliphatic oligopeptides by bridged and metallobridged bis(beta-cyclodextrin)s bearing an oxamido bis(2-benzoic) carboxyl linker. AB - beta-Cyclodextrin dimers bearing an oxamido bis(2-benzoic) carboxyl linker (1) or its metal complexes (2 and 3) were newly synthesized, and their inclusion complexation behavior with a series of representative aliphatic oligopeptides, i.e., Leu-Gly, Gly-Leu, Gly-Pro, Glu-Glu, Gly-Gly, Gly-Gly-Gly, and Glu(Cys-Gly), was elucidated by means of UV/vis, circular dichroism, fluorescence, and 2D NMR spectroscopy in Tris-HCl buffer solution (pH 7.4) at 25 degrees C. The results obtained indicated that metallobridged bis(beta-cyclodextrin)s 2 or 3 could significantly enhance the original molecular binding abilities of parent bis(beta cyclodextrin) 1 toward model substrates through the cooperative binding of two cyclodextrin moieties and the additional chelation effect supplied by the coordinated metal centers. It is interesting that hosts 2 and 3 displayed an entirely different fluorescence behavior upon complexation with guest oligopeptides. Among the guest peptides examined, 3 showed the highest complex formation constant of 68 200 M(-)(1) for Glu-Glu, up to 510-fold as compared with 1 (135 M(-)(1)), while 1 gave excellent molecular selectivity for Glu(Cys Gly)/Glu-Glu pair, up to 51-fold. The molecular binding ability and selectivity were discussed from the viewpoints of the induced-fit and multiple recognition mechanism between host and guest. PMID- 15546190 TI - Protein cargo delivery properties of cell-penetrating peptides. A comparative study. AB - Application of cell-penetrating peptides for delivering various hydrophilic macromolecules with biological function into cells has gained much attention in recent years. We compared the protein transduction efficiency of four cell penetrating peptides: penetratin, Tat peptide, transportan, and pVEC and studied the effects of various medium parameters on the uptake. Depletion of cellular energy and lowering of temperature strongly impaired the internalization of protein complexed with cell-penetrating peptides, confirming the endocytotic mechanism of peptide-mediated protein cellular transduction. Peptide-induced protein association with HeLa cells decreased 3-6-fold in energy-depleted cells. Inhibition of clathrin-dependent endocytosis by the hyperosmolar medium decreased the uptake of peptide-avidin complexes 1.5-3-fold and the removal of cholesterol from the plasma membrane 1.2-2-fold, suggesting that both clathrin-dependent and independent endocytosis were involved in peptide-induced cellular delivery of avidin. However, even under conditions of cellular energy depletion, ceasing of cellular traffic, and partial depolarization of plasma membrane, peptide-protein complexes associated with HeLa cells, as observed by FACS analysis and spectrofluorimetry. Among the studied peptides, pTat and transportan revealed higher protein transduction efficiency than penetratin or pVEC. PMID- 15546191 TI - Acetals as pH-sensitive linkages for drug delivery. AB - pH-Sensitive linkages designed to undergo hydrolysis at mildly acidic pH can trigger the release of therapeutics selectively at targets such as tumor and inflammatory tissues and in the endosomes and lysosomes of cells. Acetals have the potential to be used as linkages for a range of alcohol functionalities, and, by altering their chemical structure, it is possible to tune their hydrolysis rate. The syntheses of four conjugates of model drug molecules with PEO using acetals of varying chemical structure are described herein. Primary and secondary alcohols, as well as syn-1,2-diols, were incorporated in the conjugates. The hydrolysis kinetics were investigated by HPLC, and the conjugates had half-lives ranging from less than 1 min to several days at pH 5.0, with slower hydrolysis at pH 7.4 in all cases. These acetal linkages are therefore promising for use in a variety of drug delivery applications ranging from polymer-drug conjugates to pH sensitive micelles and nanoparticulate systems. PMID- 15546192 TI - Site-specifically traced drug release and biodistribution of a paclitaxel antibody conjugate toward improvement of the linker structure. AB - Tumor-directed drug delivery is a promising strategy in cancer treatment, and in this field, monoclonal antibodies constitute an important class of targeting vehicles. A critical issue in the design of targeting conjugates is the timing of the release of the cytotoxic payload, with the ideal situation being the release at the maximum tumor uptake of the targeting molecule. A site-specific radiolabeling technique was used to elucidate the biodistribution and in vivo drug release pattern of an antibody conjugate of paclitaxel (PTX, 1, Figure 1) in which the drug and the antibody moieties were connected by a succinate (SX) linker. In this new method, a metabolite of PTX, 3'-(4-hydroxyphenyl)paclitaxel (3'-OH-PTX, 2, Figure 1) was used as a tyrosine mimic for the synthesis of the drug site-labeled conjugate (DSL, [(125)I]-3'-OH-PTXSXC225). This was achieved by iodogen (125)I-labeling of 3'-OH-PTXSX and subsequent conjugation to C225. The antibody site-labeled conjugate (ASL, PTXSX-[(125)I]-C225) was prepared by direct radioiodination of PTXSXC225. Biodistribution of these compounds was studied in Balb/c nude mice bearing DU-145 human prostate carcinoma xenografts. While the 4 and 24 h tumor uptake (in percent injected dose per gram of tissue, %ID/g) for [(125)I]-3'-OH-PTXSXC225 were 3.3 +/- 1.5 and 1.7 +/- 0.6%ID/g, the PTXSX [(125)I]-C225 showed tumor uptake values of 3.8 +/- 4.2 and 14.8 +/- 4.2%ID/g at these time points. This difference in the tumor uptake over time indicates an early cleavage of the drug with respect to the antibody tumor localization. This was further confirmed by an in vitro drug release kinetics study leading to a half-life of about 2 h for PTXSXC225 under physiological conditions. To increase the stability of the PTX-MAb bond, a new conjugate (PTXGLC225) with glutaric acid (GL) as the linker was synthesized. Under the same conditions, the PTXGLC225 showed a 16-fold increase in the half-life (t(1/2)) of the drug release. The effect of the increased t(1/2) of this compound on the antitumor activity of the conjugate was tested in a DU-145 human prostate tumor-implanted mouse model. In comparison to a previous similar experiment with PTXSXC225, better antitumor activity was observed for the PTXGLC225 conjugate as compared to controls. These results demonstrated the first time use of radioiodinated 3'-OH-PTX for in vivo tracing of a paclitaxel conjugate and application of the resulting information to the design of a therapeutically more useful PTX-MAb linker. PMID- 15546193 TI - Sequence-selective cleavage of oligoribonucleotides by 3d transition metal complexes of 1,5,9-triazacyclododecane-functionalized 2'-O-methyl oligoribonucleotides. AB - 2'-O-Methyl oligoribonucleotides bearing a 3'-[2,6-dioxo-3,7-diaza-10-(1,5,9 triazacyclododec-3-yl)decyl phospate conjugate group have been shown to cleave in slight excess of Zn(2+) ions complementary oligoribonucleotides at the 5'-side of the last base-paired nucleotide. The cleavage obeys first-order kinetics and exhibits turnover. The acceleration compared to the monomeric Zn(2+) 1,5,9 triazacyclododecane chelate is more than 100-fold. In addition, 2'-O-methyl oligoribonucleotides having the 1,5,9-triazacyclododec-3-yl group tethered to the anomeric carbon of an intrachain 2-deoxy-beta-d-erythro-pentofuranosyl group via a 2-oxo-3-azahexyl, 2,6-dioxo-3,7-diazadecyl, or 2,9-dioxo-3,10-diazatridecyl linker have been studied as cleaving agents. These cleave as zinc chelates a tri- and pentaadenyl bulge opposite to the conjugate group approximately 50 times as fast as the monomeric chelate and show turnover. The cleavage rate is rather insensitive to the length of linker. Interestingly, a triuridyl bulge remains virtually intact in striking contrast to a triadenyl bulge. Evidently binding of the zinc chelate to a uracil base prevents its catalytic action. Replacement of Zn(2+) with Cu(2+) or Ni(2+) retards the cleaving activity of all the cleaving agents tested. PMID- 15546194 TI - Acid-degradable particles for protein-based vaccines: enhanced survival rate for tumor-challenged mice using ovalbumin model. AB - Acid-degradable protein-loaded polymer particles show promise for antigen-based vaccines due to their ability to activate cytotoxic T lymphocytes (CTLs) in vitro. Protein loadings and cytotoxic T lymphocyte activation efficiencies have now been enhanced through novel delivery vehicle designs. In particular, the use of a more hydrophilic acid-degradable cross-linker leads to increased water dispersibility and increased protein loading efficiency for the particles. A 2.5 fold increase in protein encapsulation allows the delivery of more protein antigen to antigen presenting cells (APCs) leading to a 20-fold rise in antigen presentation levels. The mechanism by which APCs internalize these particles was explored using the phagocytosis inhibitor, cytochalasin B. In addition, preliminary in vivo experiments were conducted to investigate the ability of the protein-loaded particles to provide immunity against tumors in mice, and an enhanced survival rate over the use of protein alone was observed, indicating that this vaccine delivery strategy has great practical potential. PMID- 15546195 TI - Enhanced anti-rotavirus action of human cystatin C by site-specific glycosylation in yeast. AB - The cDNA encoding human cystatin C (HCC) was subjected to site-specific substitution of alanine for serine at the position 37, to obtain the Asn(35) Lys(36)-Ser(37) sequence that is a signal for asparagine-linked (N-linked) glycosylation of protein in eukaryotes, and was transformed into Pichia pastoris X33. As a result, 1.2 mg/L oligomannosyl HCC with a carbohydrate chain of Man(10)GlcNAc(2) was produced by the Pichia transformant. The oligomannosyl HCC was more stable at the low ionic strength condition of 50 mM potassium phosphate buffer, pH 7.0, than the wild-type. In addition, the oligomannosylation substantially improved the molecular stability of cystatin against an aspartic proteinase, cathepsin D, in which the susceptibility decreased to less than 50% of nonglycosylated one. The anti-rotavirus activity of HCC was substantially enhanced by the site-directed glycosylation using the yeast expression system. A MA-104 cell line was used as a host cell for human rotavirus type-2 Wa strain in this study, to which both the wild-type and oligomannosyl HCCs did not show cytotoxicity at a concentration of 100 mug/mL. More than 80% viability of the host cell infected with 1.0 x 10(5) PFU/mL of rotavirus was conserved under the condition coexisting with 75 mug/mL of the oligomannosyl HCC, which was 15.2% higher than that of wild-type HCC. Thus, the in vitro anti-rotavirus assay indicated that the supplement of a proper amount of the oligomannosyl HCC could be used as an anti-rotavirus agent. PMID- 15546196 TI - Reversible photocontrol of peptide helix content: adjusting thermal stability of the cis state. AB - Cross-linking reagents based on an azobenzene core can be used to reversibly photoregulate secondary structure when introduced as intramolecular bridges in peptides and proteins. Photoisomerization of the azobenzene core in the trans to cis direction is triggered by photon absorption but isomerization from cis to trans occurs thermally as well as photochemically. The rate of the thermal process effectively determines the half-life of the cis form as well as the extent to which the trans form can be recovered. We designed and characterized a series of methanethiosulfonate (MTS)-bearing thiol-reactive azo-benzene-based cross-linkers. These cross-linkers are shown to permit photoregulation of helix content in a test peptide with half-lives for the cis conformation ranging from 11 s to 43 h at 25 degrees C. The cross-linkers described here thus broaden the range of reagents available for reversible photocontrol of peptide and protein conformation. PMID- 15546197 TI - Prediction of the viscosity radius and the size exclusion chromatography behavior of PEGylated proteins. AB - Size exclusion chromatography (SEC) was used to determine the viscosity radii of equivalent spheres for proteins covalently grafted with poly(ethylene glycol) (PEG). The viscosity radius of such PEGylated proteins was found to depend on the molecular weight of the native protein and the total weight of grafted PEG but not on PEG molecular weight, or PEG-to-protein molar grafting ratio. Results suggest grafted PEG's form a dynamic layer over the surface of proteins. The geometry of this layer results in a surface area-to-volume ratio approximately equal to that of a randomly coiled PEG molecule of equivalent total molecular weight. Two simple methods are given to predict the viscosity radius of PEGylated proteins. Both methods accurately predicted (3% absolute error) the viscosity radii of various PEG-proteins produced using three native proteins, alpha lactalbumin (14.2 kDa MW), beta-lactoglobulin dimer (37.4 kDa MW), and bovine serum albumin (66.7 kDa MW), three PEG reagents (2400, 5600, and 22500 MW), and molar grafting ratios of 0 to 8. Accurate viscosity radius prediction allows calculation of the distribution coefficient, K(av), for PEG-proteins in SEC. The suitability of a given SEC step for the analytical or preparative fractionation of different PEGylated protein mixtures may therefore be assessed mathematically. The methods and results offer insight to several factors related to the production, purification, and uses of PEGylated proteins. PMID- 15546198 TI - Peptide nucleic acid (PNA) amphiphiles: synthesis, self-assembly, and duplex stability. AB - Peptide amphiphiles comprising a class of conjugates of peptide nucleic acid (PNA), natural amino acids, and n-alkanes were synthesized and studied. These PNA amphiphiles (PNAA) self-assemble at concentrations between 10 and 50 muM and exhibit water solubilities above 500 muM. The highly specific, stable DNA binding properties of PNAs are preserved by these modifications, with no significant differences between the thermodynamics of DNA binding of the PNA peptide and the PNA amphiphile. Proper solubilization of the PNAA required the attachment of (Lys)(2) and (Glu)(4) peptides to PNAs, which affected the PNAA-DNA duplex stability by electrostatic interactions between these charged amino acids and the negatively charged DNA backbone. These electrostatic effects did not affect the specificity of DNA binding, however. Electrostatic effects are screened with added salt, in a manner consistent with previous studies of PNA-DNA duplex stability and predictions from a charged-cylinder model for the duplex. PMID- 15546199 TI - Synthesis of poly(ethylene glycol)-based saquinavir prodrug conjugates and assessment of release and anti-HIV-1 bioactivity using a novel protease inhibition assay. AB - Various poly(ethylene glycol)(PEG)-based prodrug conjugates of the HIV-1 protease inhibitor (PI) saquinavir (SQV) were prepared using several types of chemical groups potentially capable of modifying its pharmacokinetic properties. These prodrug conjugates included SQV-cysteine-PEG3400, SQV-cysteine-PEG3400-biotin, SQV-cysteine(R.I.CK-Tat9) [a cationic retro-inverso-cysteine-lysine-Tat nonapeptide]-PEG3400, and SQV-cysteine(R.I.CK(stearate)-Tat9)-PEG3400. SQV was linked to cysteine to form a releasable SQV-cysteine ester bond in all of the conjugates. The amino group of the cysteine moiety provided an attachment site for a slower-degrading amide bond with N-hydroxysuccinimide-activated forms of PEG- and PEG-biotin. Disulfide bonds were used to attach the cationic peptides, R.I.CK-Tat9 and R.I.CK(stearate)-Tat9 to the cysteine moiety in order to provide cell-specific release. An assay was established and validated for measuring the activity of SQV and other protease inhibitors in biological samples. In this assay, cleavage of an internally quenched fluorescent substrate, Arg-Glu(EDANS) Ser-Gln-Asn-Tyr-Pro-Ile-Val-Gly-Lys(DABCYL)-Arg by HIV-1 protease was inhibited by SQV in a dose-dependent manner at concentrations of 0.05-0.5 microM. All prodrug conjugates were shown to be inactive in this assay until the ester bond was cleaved and active SQV was released. The prodrug reconversion half-lives in 0.1 N HCl, phosphate-buffered saline (PBS) at pH 7.4 and in spiked plasma at 37 degrees C were 9, 14, and 0.9 h, respectively. The anti-HIV-1 activity (ED(50)) of the PEG-based SQV prodrug conjugates was evaluated in MT-2 cells using an MTT assay. The activity of conjugated SQV was reduced (ED(50) = 900 nM) for the PEG only conjugate, but restored with the addition of biotin (ED(50) = 125 nM), R.I.CK-Tat9 (ED(50) = 15 nM), and R.I.CK(stearate)-Tat9 (ED(50) = 62 nM) as compared to maximum achievable anti-HIV-1 activity (unconjugated SQV, control, ED(50) = 15 nM), suggesting enhanced cellular uptake of conjugates. Cytotoxicity (LD(50)) was assessed for all prodrug conjugates using non-HIV-1 infected cells and was found to be in the micromolar range. The difference between the LD(50) and ED(50) suggests a favorable therapeutic index for the prodrug conjugates. In conclusion, these promising initial results demonstrate that the reconversion of the conjugate prodrugs was complete and that active SQV was released. Since the major delivery advantages of PEG prodrug conjugates can only be observed in vivo, issues of reconversion and elimination half-lives in plasma will have to be further studied in an in vivo model. The current results also demonstrate that the protease inhibition assay is a simple yet effective bioanalytical tool that can be used to assess the release and anti-HIV-1 activity of HIV-1 PIs from their prodrug forms. PMID- 15546200 TI - Novel NMR platform for detecting gene transfection: synthesis and evaluation of fluorinated phenyl beta-D-galactosides with potential application for assessing LacZ gene expression. AB - Gene therapy holds great promise for the treatment of diverse diseases, but widespread implementation is hindered by difficulties in assessing the success of transfection. The development of noninvasive reporter techniques based on appropriate molecules and imaging modalities may help to assay gene expression. Fluorophenyl-beta-d-galactopyranosides provide a novel class of NMR active molecules, which are highly responsive to the action of beta-galactosidase (beta gal), the product of the lacZ gene. The reporter molecules are stable in solution and with respect to wild-type cells, but the enzyme causes liberation of the aglycon, a fluorophenol, accompanied by distinct color formation and a (19)F NMR chemical shift of 5-10 ppm, depending on pH. Synthetic strategy, experimental methods, and molecular and (19)F NMR characteristics are reported for a series of molecules in solution, blood, and tumor cells. This class of molecules presents a new strategy for assaying gene expression with a highly versatile molecular structural platform. PMID- 15546201 TI - DNA complexing lipopolythiourea. AB - We present a neutral lipopolythiourea (DTTU) as a potential DNA-binding agent. Light scattering experiments showed that mixing a lipopolythiourea with dipalmitoylphosphatidylcholine (DPPC/DTTU) led to small particles with sizes ranging from 100 to 150 nm at optimum conditions. Setting a fixed DNA amount, an increasing amount of DTTU/DPPC or DPPC lipids was added. Particle size increased only with DTTU/DPPC, indicating that interaction occurred between the DTTU/DPPC particles and DNA. In the same way, only DTTU/DPPC limited the ethidium bromide accessibility to plasmid DNA. These data suggest that DTTU/DPPC liposomes associate to DNA, which was confirmed by agarose gel experiments. To prove the active part of the DTTU lipid itself in DNA compaction, pegoylated-lipid was used. Cholesterol-PEG(2000) alone was not able to condense DNA. In contrast, DTTU/PEG-cholesterol was able to retain plasmid DNA on an agarose gel. In vivo injection of DTTU/DPPC/complexes was studied. Circulation time increase for noncationic particles as compared to cationic. More obvious was the lack of nonspecific accumulation in the lung, where a gain of 3 to 40 fold was measured. PMID- 15546202 TI - Synthesis and biological activity of water-soluble maleimide derivatives of the anticancer drug carboplatin designed as albumin-binding prodrugs. AB - Four platinum (II) complexes (13-16) were synthesized by reacting either [Pt trans-DACH](NO(3))(2) with a 6-maleimidocaproic acid, a 15-maleimido-4,7,10,13 tetroxapentadecanoic acid, and a 6-maleimido-4-oxacaproic ester derivative of cyclobutane-1,1-dicarboxylic acid (CDBA) or [Pt(NH(3))(2)](NO(3))(2) with a 6 maleimido-4-oxacaproic ester derivative of CBDA. Both complexes containing the 6 maleimido-4-oxacaproic ester (15, 16) showed good water solubility (>/=8 mg/mL) and CE experiments revealed rapid binding to human serum albumin and the formation of biadducts with dGMP and dAMP. In the MaTu xenograft model in nude mice, both complexes showed an improved antitumor effect at their maximum tolerated dose (2 x 50 mg/kg carboplatin equivalents) compared to therapy with carboplatin at equimolar dose or at its optimal dose (2 x 75 mg/kg). PMID- 15546203 TI - Design of metalloporphyrin-carbohydrate conjugates for a new superoxide dismutase mimic with cellular recognition. AB - Metalloporphyrin-carbohydrate conjugates have been synthesized as superoxide dismutase (SOD) mimics with cellular recognition. To synthesize the conjugates, aliphatic primary amino groups for conjugation were introduced, with the cationic pyridyl groups for the SOD activity of porphyrin preserved. The reductive amination between introduced amino groups and the reducing end of lactose was then carried out. The resulting conjugates consisting of manganese (Mn)-porphyrin surrounded by several lactose molecules possessed significant SOD activity and low cytotoxicity. Compared with metalloporphyrins having no lactose molecule, the recognition of the resulting conjugates by human hepatoma HepG2 cells increased. The cellular recognition was inhibited by competitors of beta-galactose. These results suggest that the Mn-porphyrin-lactose conjugates recognized the hepatic lectin on the cell surface. PMID- 15546204 TI - In vitro cytotoxicity and in vivo distribution after direct delivery of PEG camptothecin conjugates to the rat brain. AB - Low water solubility and rapid elimination from the brain inhibits local delivery via implants and other delivery systems of most therapeutic drugs to the brain. We have conjugated the chemotherapy drug, camptothecin (CPT), to poly(ethylene glycol) (PEG) of molecular weight 3400 using previously established protocols. These new conjugates are very water-soluble and hydrolyze at a pH-dependent rate to release the active parent drug. We have studied the uptake of these conjugates by cells in vitro and quantified their cytotoxicity toward gliosarcoma cells. These conjugates were loaded into biodegradable polymeric controlled-release implants, and their release characteristics were studied in vitro. We implanted similar polymeric disks into rat brains and used a novel sectioning scheme to determine the concentration profile of CPT in comparison to conjugated CPT in the brain after 1, 7, 14, and 28 days. We have found that PEGylation greatly increases the maximum achievable drug concentration and greatly enhances the distribution properties of CPT, compared to corelease of CPT with PEG. Although only one percent of CPT in the conjugate system was found in the hydrolyzed, active form, drug concentrations were still significantly above cytotoxic levels over a greater distance for the conjugate system. On the basis of these results, we believe that PEGylation shows great promise toward increasing drug distribution after direct, local delivery in the brain for enhanced efficacy in drug treatment. PMID- 15546205 TI - Multimodality in vivo molecular-genetic imaging. AB - Multimodality imaging is increasingly being used in molecular-genetic studies in small animals. The coupling of nuclear and optical reporter genes represents the beginning of a far wider application of this technology. Optical imaging and optical reporter systems are cost-effective and time-efficient, they require less resources and space than PET or MRI, and they are particularly well suited for small animal imaging and for in vitro assays to validate different reporter systems. However, optical imaging techniques are limited by depth of light penetration and scatter and do not yet provide optimal quantitative or tomographic information. These issues are not limiting for PET- or MRI-based reporter systems, and PET- and MRI-based animal studies are more easily generalized to human applications. Many of the shortcomings of each modality alone can be overcome by the use of dual- or triple-modality reporter constructs that incorporate the opportunity for PET, fluorescence and bioluminescence imaging. We optimistically expect that some form of tomographic, small animal optical imaging capability will be developed soon, and that this will provide the opportunity for the colocalization of optical signals to anatomical structures provided by tomographic CT and MR imaging. PMID- 15546206 TI - Converting weak binders into infinite binders. AB - Monoclonal antibody 2D12.5 binds DOTA chelates of all the rare earths with K(d) approximately 10(-)(8) M, making it useful for the capture of probe molecules with a variety of properties. To make 2D12.5 even more useful for biological applications, we have engineered a single cysteine residue at position 54 of the heavy chain, a site proximal to the protein's binding site, so that weakly electrophilic metal complexes of (S)-2-(4-acrylamidobenzyl)-DOTA (AABD) may bind and form permanent linkages. At 37 degrees C, pH 7.5, all of the rare earth-AABD complexes bind permanently to the 2D12.5 G54C mutant within 5 min, in yields that correlate with their relative binding affinities. Surprisingly, indium-AABD also binds permanently in >50% yield within 5 min, despite the fact that changing the metal to indium reduces the affinity approximately 100x; even copper-AABD, which has approximately 10 000x lower binding affinity than the rare earths, binds permanently in >70% yield within 2 h. However, acrylamido compounds with no measurable affinity do not bind permanently. The important practical implication is that the G54C mutant of 2D12.5 may be used for applications that include not only the rare earths, but also an unexpected range of other elements as well. This infinite binding system can exhibit selective and permanent attachment with a remarkable range of structurally related ligands, albeit at slower rates as affinities decrease. PMID- 15546207 TI - Irreversible engineering of the multielement-binding antibody 2D12.5 and its complementary ligands. AB - Engineering the permanent formation of a receptor-ligand complex has a number of potential applications in chemistry and biology, including targeted medical imaging and therapy. Starting from the crystal structure of the rare-earth-DOTA binding antibody 2D12.5 (Corneillie, T. M., Fisher, A. J., and Meares, C. F. (2003) J. Am. Chem. Soc. 125, 15039-15048), we used the site-directed incorporation of cysteine nucleophiles at the periphery of the antibody's binding site, paired with the chemical design of a weakly electrophilic ligand, to produce a receptor-ligand pair that associates efficiently and permanently. Protein residues proximal to the ligand's side chain were identified for engineering cysteine mutants. Fab fragments incorporating a cysteine at position 54, 55, or 56 of the heavy chain (complementarity determining region 2) were designed from the structure and then cloned, expressed in Drosophila S2 cells, and tested for reactivity with mildly electrophilic DOTA-yttrium ligands. All showed permanent binding activity, indicating that there is some tolerance for the location of the reactive mutant on the protein surface near the binding site. The G54C Fab mutant displayed the highest expression levels and permanent binding activity in initial experiments and was produced in high yield for further study. Upon examining the behavior of the G54C mutant with a small set of electrophilic ligands, differences in reactivity were observed which indicated that the substituents near the electrophilic atom can be important determinants of permanent binding. The G54C mutant permanently attaches to Y(3+) complexes of (S) 2-(4-acrylamidobenzyl)-DOTA with a half-time of approximately 13 min at 37 degrees C, making it potentially useful for in vivo pretargeting applications. PMID- 15546208 TI - Developing a peptide-based near-infrared molecular probe for protease sensing. AB - Recently near-infrared (NIR) molecular probes have become important reporter molecules for a number of types of in vivo biomedical imaging. A peptide-based NIR fluorescence probe consisting of a NIR fluorescence emitter (Cy5.5), a NIR fluorescence absorber (NIRQ820), and a protease selective peptide sequence was designed to sense protease activity. Using a MMP-7 model, we showed that NIRQ820 efficiently absorbs the emission energy of Cy5.5 resulting in a low initial signal. Upon reacting with its target, MMP-7, the fluorescence signal of the designed probe was increased by 7-fold with a K(cat)/K(m) of 100 000 M(-)(1) s( )(1). The described synthetic strategy should have wide application for other NIR probe preparations. PMID- 15546209 TI - Synthesis and characterization of poly(L-glutamic acid) gadolinium chelate: a new biodegradable MRI contrast agent. AB - Most currently evaluated macromolecular contrast agents for magnetic resonance imaging (MRI) are not biodegradable. The goal of this study is to synthesize and characterize poly(l-glutamic acid) (PG) gadolinium chelates as biodegradable blood-pool MRI contrast agents. Two PG chelates of gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) were synthesized through the use of difunctional and monofunctional DTPA precursors. The conjugates were characterized with regard to molecular weight and molecular weight distribution, gadolinium content, relaxivity, and degradability. Distributions of the polymeric MRI contrast agents in various organs were determined by intravenous injection of (111)In-labeled polymers into mice bearing murine breast tumors. MRI scans were performed at 1.5 T in mice after bolus injection of the polymeric chelates. PG Hex-DTPA-Gd, obtained from aminohexyl-substituted PG and DTPA-dianhydride, was partially cross-linked and was undegradable in the presence of cathepsin B. On the other hand, PG-Bz-DTPA-Gd synthesized directly from PG and monofunctional p aminobenzyl-DTPA(acetic acid-tert-butyl ester) was a linear polymer and was degradable. The relaxivities of the polymers at 1.5 T were 3-8 times as great as that of Gd-DTPA. Both polymers had high blood concentrations and were primarily accumulated in the kidney. However, PG-Bz-DTPA-Gd was gradually cleared from the body and had significantly less retention in the blood, the spleen, and the kidney. MRI with PG-Bz-DTPA-Gd in mice showed enhanced vascular contrast at up to 2 h after the contrast agent injection. The ability of PG-Bz-DTPA-Gd to be degraded and cleared from the body makes it a favorable macromolecular MRI contrast agent. PMID- 15546210 TI - A new high affinity technetium analogue of bombesin containing DTPA as a pharmacokinetic modifier. AB - The bombesin (BN)/gastrin-releasing peptide (GRP) receptor is expressed in high density on the cell surface of a variety of tumors. This makes the receptors accessible as a molecular target for the detection of lesions in which they are expressed. In this study, we describe a high affinity hydrophilic (99m)Tc-labeled BN analogue, [DTPA(1), Lys(3)((99m)Tc-Hx-DADT), Tyr(4)]BN, having diethylenetriaminepentaacetic acid (DTPA), as a build-in pharmacokinetic modifier, to direct its excretion through the urinary system in order to lower abdominal background activity. In vitro binding studies using [(125)I-Tyr(4)]BN (K(d), 0.1 nM) and human prostate cancer PC-3 cell membranes showed that the inhibition constant (K(i)) of [DTPA(1), Lys(3)((99)Tc-Hx-DADT), Tyr(4)]BN was 19.9 +/- 8.0 nM. Biodistribution studies in normal mice showed fast blood clearance (0.15 +/- 0.01% ID/g, 4 h postinjection), low intestinal accumulation (9.16 +/- 2.35% ID/g, 4 h postinjection), and significant uptake in BN/GRP receptor rich tissues such as the pancreas (21.83 +/- 2.88% ID/g, 15 min postinjection). The pancreas/blood, pancreas/muscle, and pancreas/liver ratios were highest at 2 h postinjection at 23, 74, and 8.4, respectively. The uptake in the pancreas could be blocked by BN (11.96 +/- 1.17 vs 0.65 +/- 0.16% ID/g), partially blocked by neuromedin B (11.96 +/- 1.17 vs 6.66 +/- 0.51% ID/g), but not affected by somatostatin (11.96 +/- 1.17 vs 12.91 +/- 2.53% ID/g), indicating that the binding of [DTPA(1), Lys(3)((99m)Tc-Hx-DADT), Tyr(4)]BN to the receptors was specific. Scintigraphic imaging of human PC-3 prostate cancer xenografts in SCID mice gave a high target to nontarget ratio on the image. Thus, [DTPA(1), Lys(3)((99m)Tc-Hx-DADT), Tyr(4)]BN has the potential for imaging BN/GRP receptor positive lesions. PMID- 15546211 TI - PEG-g-poly(GdDTPA-co-L-cystine): a biodegradable macromolecular blood pool contrast agent for MR imaging. AB - Biodegradable PEGylated Gd-DTPA l-cystine copolymers, PEG-g-poly(GdDTPA-co-l cystine), were prepared and tested as a blood pool contrast agent in mice. The biodegradable macromolecular agent was designed to be broken down into smaller Gd complexes by endogenous thiols via the disulfide-thiol exchange reaction to facilitate the clearance of Gd complexes after the contrast-enhanced MRI examination. Gd-DTPA l-cystine copolymers were synthesized by condensation polymerization of l-cystine and DTPA-dianhydride in water followed by chelating with Gd(OAc)(3). MPEG-NH(2) (MW = 2000) was then conjugated to the polymeric backbone in different ratios. The macromolecular contrast agent was readily degraded with the incubation of l-cysteine. It also demonstrated superior contrast enhancement in the heart and blood vessels as compared to a low molecular weight control agent, Gd-(DTPA-BMA). At 1 h postcontrast, the PEGylated macromolecular agent still showed prominent enhancement, while little contrast enhancement was detectable in the blood pool by the control agent. PEG-g poly(GdDTPA-co-l-cystine) shows promise as an MR blood pool imaging agent. PMID- 15546212 TI - Cyclen-based phenylboronate ligands and their Eu3+ complexes for sensing glucose by MRI. AB - Novel cyclen-based phenylboronate ligands and their corresponding Eu(3+) complexes have been examined as glucose sensors using chemical exchange saturation transfer (CEST) MR imaging for detection. Two isomeric bis phenylboronate complexes, Eu(4) and Eu(10), and a mono-phenylboronate complex, Eu(12), had been prepared and characterized by UV and circular dichroism spectroscopy, mass spectrometry, and CEST imaging. Both the free ligands and their Eu(3+) complexes bind to simple sugars, but their selectivity and binding affinities vary with sugar structure. Interestingly, the free ligands, 4 and 10, are selective for fructose over glucose, but this selectivity order switches in the respective Eu(3+) complexes. Of the complexes examined, Eu(4) shows the highest selectivity and binding affinity for glucose (2275 +/- 266 M(-)(1) at pH 10.2 and 339 +/- 29 M(-)(1) at pH 7). Glucose acts as a "capping"moiety in the Eu(4).glucose binary complex and modulates water exchange between a single Eu(3+) bound water molecule and bulk water, an effect that can be detected by CEST imaging. Thus, Eu(4) represents a new class of metabolite-specific imaging agents that may allow mapping of metabolites by MRI of the bulk water signal. PMID- 15546213 TI - Preparation and properties of 99mTc(CO)3+-labeled N,N-bis(2-pyridylmethyl)-4 aminobutyric acid. AB - Labeling biomolecules with (99m)Tc(CO)(3)(+) ((99m)Tc tricarbonyl) is attracting increasing attention. Although histidine is often considered an ideal bifunctional chelator for (99m)Tc (or (188)Re) tricarbonyl, the family of dipicolylamine carboxylate chelators may be a useful alternative because of the expected ease of synthesis and because the structure provides a pendent carboxylate for potential conjugation to biomolecules. The dipicolylamine chelator N,N-bis(2-pyridylmethyl)-4-aminobutyric acid (BPABA) was synthesized using 4-aminobutyric acid in place of glycine or aminopropionic acid in the literature, to avoid possible involvement of the carboxylate in the complex formation process by forming five- or six-membered chelation rings. Using a commercial tricarbonyl kit (Mallinckrodt), the complex formation properties of both BPABA and commercial histidine with (99m)Tc tricarbonyl were investigated, and the in vitro complex stabilities in saline and in serum were compared. Stability in vivo was also examined following i.v. administration to normal mice. BPABA was synthesized simply and quantitatively by reacting picolyl chloride with aminobutyric acid in one step. On RP HPLC, the product eluted essentially in one peak and the structure was confirmed by ESI-MS. After labeling, both BPABA and histidine were shown by RP HPLC to form tricarbonyl complexes. In both cases, after incubation at 100 degrees C for 20 min, only one predominant peak of (99m)Tc(CO)(3)(+)-histidine or (99m)Tc(CO)(3)(+)-BPABA was apparent, and both complexes were stable at room temperature in saline for at least 24 h. After incubation for 24 h in 37 degrees C serum, by SE HPLC, 20% of the (99m)Tc(CO)(3)(+)-histidine was bound to serum protein compared to less than 10% for (99m)Tc(CO)(3)(+)-BPABA. A 5000 molar excess of histidine at 100 degrees C for 6 h was unable to dissociate (99m)Tc(CO)(3)(+)-BPABA. By contrast, BPABA easily dissociated (99m)Tc(CO)(3)(+)-histidine under the same conditions. Both complexes were stable in vivo in mice, and (99m)Tc(CO)(3)(+)-BPABA showed rapid and specific hepatobiliary clearance while (99m)Tc(CO)(3)(+)-histidine was cleared through the kidneys. In conclusion, BPABA was easily synthesized and was shown to possess properties comparable to histidine for labeling of biomolecules with (99m)Tc tricarbonyl. However, it was found that the chelator concentration required for quantitative (99m)Tc tricarbonyl labeling with both BPABA and histidine were 2 orders higher than that required with more conventional labeling using MAG(3). Finally, the complex (99m)Tc(CO)(3)(+)-BPABA itself was found to clear exclusively via the hepatobiliary pathway and may have value as a potential hepatobiliary imaging agent. PMID- 15546214 TI - 18F-fluorothiols: a new approach to label peptides chemoselectively as potential tracers for positron emission tomography. AB - [(18)F]Fluorothiols are a new generation of peptide labeling reagents. This article describes the preparation of suitable methanesulfonyl precursors and their use in no-carrier-added radiosyntheses of (18)F-fluorothiols. The preparations of (3-[(18)F]fluoropropylsulfanyl)triphenylmethane, (2-[2-[2-(2 [(18)F]fluoroethoxy)ethoxy]ethoxy]ethylsulfanyl)triphenylmethane, and 4 [(18)F]fluoromethyl-N-[2-triphenylmethanesulfanyl)ethyl]benzamide starting from the corresponding methanesulfonyl precursors were investigated. Following the removal of the triphenylmethane protecting group, the (18)F-fluorothiols were reacted with the N-terminal chloroacetylated model peptide ClCH(2)C(O) LysGlyPheGlyLys. The corresponding radiochemical yields of (18)F-labeled isolated model peptide, decay-corrected to (18)F fluoride, were 10%, 32%, and 1%, respectively. These results indicate a considerable potential of (18)F fluorothiols for the chemoselective labeling of peptides as tracers for positron emission tomography (PET). PMID- 15546215 TI - Streptavidin in antibody pretargeting. 4. Site-directed mutation provides evidence that both arginine and lysine residues are involved in kidney localization. AB - The in vivo application of the protein streptavidin is limited by its propensity to localize to kidney, particularly when it is used as a carrier of radionuclides in Targeted Radionuclide Therapy. Our previous studies demonstrated that modification of recombinant "core" streptavidin (rSAv) by reaction of lysine residues with succinic anhydride and arginine residues with 1,2-cyclohexanedione dramatically decreases the kidney concentrations over that obtained with wild type rSAv. In this investigation, we explored the role of lysine and arginine residues in kidney localization further by evaluating site-directed mutants of rSAv. In the five mutants studied, the four lysine residues found in each subunit of rSAv were replaced (independently) with an alanine (K80A, K121A, K132A, K134A), and a specific arginine was replaced with a histidine (R59H). The rSAv mutants were prepared from a "core" rSAv construct produced by expression in E. coli that had 124 amino acids (residues 13-136). Another rSAv construct that had 127 amino acids (residues 13-139), used in most of our previous studies, was also included for comparison. As an additional comparison, succinylated rSAv was prepared and evaluated. The rSAv proteins were radioiodinated and injected into athymic mice that were on a biotin-free diet for 5-7 days prior, and biodistribution data were obtained (for most proteins) at 1, 4, 24, and 48 h postinjection. The data obtained show large differences in kidney localizations of the wild-type rSAv and some rSAv mutants. The largest difference in the kidney concentration was noted for the rSAv-K134A mutant (1.90 +/- 0.22%ID/g; 24 h pi) as compared to the wild-type rSAv (31.83 +/-5.26%ID/g) at the same time point. The concentration of rSAv-K134A mutant in kidney was slightly lower than that obtained with succinylated rSAv. At the 24 h time point, the kidney concentrations of the rSAv-R59H mutant (8.95 +/- 2.94%ID/g) and the rSAv-K121A mutant (11.86 +/- 1.61%ID/g) were lower than wild-type rSAv, but the rSAv mutants rSAv-K80A (27.95 +/- 1.82%ID/g) and rSAv-K132A (32.50 +/- 10.09%ID/g) were essentially the same. The data suggests that specific lysine and arginine residues are involved in kidney localization. Possible mechanisms for the observed kidney localization are discussed. PMID- 15546216 TI - Radiopharmaceuticals for assessment of multidrug resistance P-glycoprotein mediated drug transport activity. AB - Multidrug resistance (MDR) mediated by overexpression of MDR1 P-glycoprotein (Pgp) is one of the best characterized transporter-mediated barriers to successful chemotherapy in cancer patients. Thus, noninvasive interrogation of Pgp-mediated transport activity in vivo would be beneficial in guiding therapeutic choices. Both small organic medicinals as well as metal complexes characterized as transport substrates for Pgp are amenable to incorporation of PET or SPECT radionuclides and may enable noninvasive imaging of Pgp in cancer patients. Toward this objective, clinically approved agents, exemplified by (99m)Tc-Sestamibi and (99m)Tetrofosmin, have already shown promise for the functional evaluation of Pgp-mediated transport activity in human tumors in vivo. In addition, selected agents from an upcoming class of substituted Schiff-base gallium(III) complexes containing an N(4)O(2) donor core in their organic scaffold and capable of generating both SPECT and PET radiopharmaceuticals have also been shown to be promising for noninvasive assessment of Pgp activity in vitro and in vivo. PMID- 15546217 TI - Evidence of antisense tumor targeting in mice. AB - Even though increased accumulations of radiolabeled antisense DNAs compared to control DNAs are becoming a routine observation in cultured tumor cells, trustworthy evidence of tumor targeting in vivo by an antisense mechanism remains elusive. The goal of this study was to obtain convincing evidence of antisense tumor targeting in nude mice by using two different tumors and both intratumoral (i.t.) and intravenous (i.v.) administration of radiolabeled antisense and control sense DNAs. Both the MDR++ cell line KB-G2 and its parent MDR+ cell line KB-31 were used in this study. The antisense (AS) DNA was directed against the AUG start codon of the MDR1 mRNA and, along with the sense (S) control DNA, was a uniform phosphorothioate administered naked. In previous cell culture studies from our laboratories, the accumulation of this AS DNA was strikingly high in KB G2 cells and only average in KB-31 cells, a fact we attribute to the 1000-fold higher expression by RT-PCR of MDR1 mRNA in the former cell line. In this study, both DNAs were radiolabeled with (99m)Tc via MAG3 and administered i.t. or i.v. at 1 microg (100 microCi) per animal 24 h prior to sacrifice and dissection in mice bearing thigh tumors of about 1 g. Following i.t. administration, no statistically significant differences (Student's t test, p < 0.05, N = 4) between the AS and S DNA biodistributions in normal tissues were observed except in the KB-G2 mice in which muscle levels were lower for the S control. In contrast, tumor levels in the KB-G2 animals were significantly higher for the AS DNA vs S DNA (14.7 vs 8.5% ID/g) while this difference (8.6 vs 4.3% ID/g) was insignificant in the KB-31 animals. The whole body images obtained just prior to sacrifice clearly show improved targeting of AS DNA vs S DNA in the KB-G2 but not the KB-31 animals. Calculations based on these results show that about 60 000 AS DNAs accumulated specifically (i.e. AS DNA - S DNA) per KB-G2 tumor cell following i.t. administration. When administered i.v. rather than i.t., higher tumor levels in KB-G2 animals compared to KB-31 were not observed, most likely because of the lower dosage reaching the tumors. When the KB-G2 and KB-31 results are combined, no statistically significant differences between the AS and S DNA biodistributions in normal tissues were observed except in blood in which S DNA levels were higher and in spleen in which they were lower. In contrast, tumor levels were significantly higher for the AS DNA vs S DNA (0.100 vs 0.063% ID/g). Calculations based on these results show that about 400 AS DNAs accumulated specifically per tumor cell following i.v. administration. Therefore evidence for tumor targeting in vivo by an antisense mechanism has been obtained in that statistically higher tumor accumulations of the (99m)Tc-AS DNA were observed compared to the control (99m)Tc-S DNA both following i.t. and i.v. administrations. The successful localization of AS DNA in tumor demonstrates that in vivo AS targeting of tumor is feasible although improvements in tumor delivery and normal tissue clearance are needed for practical antisense imaging. PMID- 15546218 TI - Synthesis and properties of fluorescent NF-kappa B-recognizing hairpin oligodeoxyribonucleotide decoys. AB - Intramolecular fluorescence quenching of cyanine dyes was investigated using a model hairpin oligonucleotide decoy encoding a NF-kappaB p50 subunit binding site. Two types of hairpin oligonucleotides were synthesized: (1) 5'-(6 aminohexyl)- and 3'-(3-aminopropyl)-linked (I); (2) 5'-(6-aminohexyl)- and 3'-[3 (3-hydroxypropyldithio)propyl]-linked (II). Oligonucleotide I was covalently modified using monofunctional either Cy3- or Cy5.5-N-hydroxysuccinimide esters. Using reverse-phase HPLC, mono-and dicyanineamide derivatives of I were isolated. Mono-Cy3-modified derivatives of I, but not the mono-Cy5.5-modified derivatives, showed a 2-fold higher Cy3 fluorescence intensity compared to the free dye. There was no detectable difference in fluorescence between the di-Cy3 derivative of I and the free dye at the same concentration. However, there was a 4-fold quenching of fluorescence in the case of the di-Cy5.5 derivative of the same hairpin oligonucleotide. The quenching of Cy5.5 fluorescence could not be explained by the interaction of Cy5.5 with nucleotide bases as demonstrated by incubating free Cy5.5 dye with oligonuclotides. The quenching effect was further investigated using an oligonucleotide bearing a cleavable 3'-amino-terminated linker bearing an S-S bond (III). After modification of the 5'- and 3'-end of oligonucleotide III with a Cy5.5 monofunctional hydroxysuccinimide ester, a 70-75% quenching of fluorescence was observed. Fluorescence was 100% dequenched after the reduction of S-S bond. The obtained result unequivocally demonstrates that the formation of intramolecular Cy5.5 dimers is the dominant mechanism of fluorescence quenching in symmetric dye-dye hairpin decoy beacons. PMID- 15546219 TI - Targeted molecular imaging agents for cellular-scale bimodal imaging. AB - Molecular imaging is a powerful tool that has the ability to elucidate biochemical mechanisms and signal the early onset of disease. Overexpression of the peripheral benzodiazepine receptor (PBR) has been observed in a variety disease states, including glioblastoma, breast cancer, and Alzheimer's disease. Thus, the PBR could be an attractive target for molecular imaging. In this paper, the authors report cellular uptake and multimodal (MRI and fluorescence) imaging of PBR-overexpressing C6 glioblastoma (brain cancer) cells using a cocktail administration approach and a new PBR targeted lanthanide chelate molecular imaging agent. PMID- 15546220 TI - Estimating the number of bound waters in Gd(III) complexes revisited. Improved methods for the prediction of q-values. AB - Two literature computational methods for the prediction of the number of inner sphere aqua ligands, q, have been applied to a test set of seven Gd(aminocarboxylate) complexes. The first method is based on the hypothesis that q should be proportional to the solvent-accessible surface area of the ligand complexed Gd ion (Castonguay, L. A., Treasurywala, A. M., Caulfield, T. J., Jaeger, E. P., and Kellar, K. E. (1999) Prediction of q-Values and Conformations of Gadolinium Chelates for Magnetic Resonance Imaging. Bioconjugate Chem. 10, 958). The second method is based on the hypothesis that q-values can be deduced by examining series of steric energy versus ionic radii plots as a function of coordination number (Reichert, D. E., Hancock, R. D., and Welch, M. J. (1996) Molecular Mechanics Investigation of Gadolinium(III) Complexes. Inorg. Chem. 35, 7013). This study identifies deficiencies in these methods and, with respect to the first method, describes some apparent errors. Although neither method was reliable at predicting q, two alternate approaches based on either molecular mechanics strain thresholds or exposed Gd surface area thresholds are shown to predict observed q-values for all Gd aminocarboxylate complexes in the test set. PMID- 15546221 TI - Targeted therapies in the treatment of colorectal cancer: what managed care needs to know. AB - OBJECTIVE: This review is designed to explore the disease, its current treatment, the expanding field of antiangiogenic treatments, and the implications of these advances for the managed care patient. DATA SOURCES: This article is based, in part, on presentations given by the authors in a continuing education symposium presented during the Academy of Managed Care Pharmacy.s 16th Annual Meeting and Showcase, April 1, 2004, in San Francisco. CONCLUSIONS: Colorectal cancer (CRC) is the third most common cancer in the United States, and the second-leading non.gender-specific cause of cancer deaths. If the cancer is caught soon enough (before node involvement and metastasis occur), there is a strong chance of survival; however, only slightly more than one third of cases are detected that soon. Emerging treatments that target only the cancer cells are increasing the length of survival for those who are diagnosed at later stages of the disease. PMID- 15546222 TI - Quality improvement in the continuum of care: impact of atherothrombosis in managed care pharmacy. AB - OBJECTIVE: To review the clinical and economic impact of atherothrombosis, the use of antiplatelet therapy for patients with atherothrombosis, and the impact of disease management programs on quality improvement and health care costs. SUMMARY: Atherothrombosis is a new term that describes the formation of a thrombus on an existing atherosclerotic plaque. While simple, this pathophysiologic mechanism underlies a vast array of vascular diseases, including myocardial infarction, ischemia, peripheral arterial disease, vascular death, and many forms of stroke. In fact, atherothrombosis is the leading cause of death worldwide. Treatment approaches include lifestyle modification and the use of pharmacologic agents such as lipid-lowering therapy, antihypertensive agents, hypoglycemic therapy, ACE inhibitors, and beta-blockers. Despite these treatments, studies have indicated that not all patients receive recommended therapies. The continuing upward spiral in health care costs in the United States has spurred numerous initiatives aimed at improving the quality of care while maintaining or reducing costs. Among these approaches to quality improvement are programs that promote adherence to evidence-based clinical data, including published guidelines, and improve the appropriate use of pharmaceuticals. Although expenditures for prescription drugs are rising, they remain a minority of overall health care expenditures, and evidence suggests that improved usage of medications can reduce other direct costs, such as hospitalizations and inpatient care, which may account for a far greater proportion of total costs. CONCLUSION: The utilization of quality improvement techniques and disease management tools can be used to improve the quality of care of patients with atherothrombosis. A key component of this strategy is pharmacologic therapy. The appropriate use of prescription drugs involves several key factors, including the proper selection of the agent based on available clinical evidence, choosing regimens that enhance patient compliance, and sound methodologies to evaluate outcomes. Improvements in the quality of care for patients with atherothrombosis can have a significant impact on the overall quality of health care as well as total costs. PMID- 15546223 TI - Written emotional disclosure buffers the effects of social constraints on distress among cancer patients. AB - The aims of the present study were to examine whether written emotional disclosure would reduce distress among cancer patients and whether it would buffer the effects of high levels of social constraint (negative social responses to patients' expressions of emotion regarding their cancer) on distress. Cancer patients (N=104) were randomly assigned to write about their emotions regarding their cancer 20 min a day for 3 days or to write about a nonemotional topic. They completed questionnaires at baseline and 6 months postintervention. Results showed that written disclosure buffered the effects of social constraints on stress at the 6-month follow-up and that avoidance partly mediated these effects. The present data reinforce the notion that interventions should be tailored to patients' needs. PMID- 15546224 TI - A longitudinal analysis of the course of depressive symptomatology in geriatric patients with cancer of the breast, colon, lung, or prostate. AB - This study mapped the trajectory of depression and its components (depressive mood, somatic expression of depression, and lack of positive affect) for 1 year after an initial cancer diagnosis, revealing the complex nature of the psychological response to the cancer experience. The analysis was based on 4 waves of panel data from 860 older patients with incident breast, colon, lung, or prostate cancer. Predictors of depressive symptoms included cancer site; stage; comorbidities; sociodemographic characteristics; and indicators of physical functioning, symptom severity, and treatment. Patients' overall depressive symptoms declined, especially depressive mood and somatic indicators. By contrast, the sense of well-being did not recover; in fact, it would have deteriorated without improvements in physical functioning and physical symptoms. The present findings show the importance of psychological assessments and symptom management during cancer treatment. PMID- 15546225 TI - Psychological and social predictors of changes in fruit and vegetable consumption over 12 months following behavioral and nutrition education counseling. AB - This study assessed psychological and social factors predicting 12-month changes in fruit and vegetable consumption achieved by 271 men and women from a low income population randomized to brief behavioral and nutrition education counseling. Greater increases in fruit and vegetable intake were achieved in the behavioral than in the nutrition education condition (1.49 vs. 0.87 portions per day, p=.021). Increases were predicted by baseline social support for dietary change but not by baseline psychological measures. However, short-term (8-week) changes in dietary self-efficacy, encouragement, anticipated regret, perceived benefits, and knowledge of recommended intake predicted 12-month changes in fruit and vegetable consumption independently of gender, age, ethnicity, income, and baseline intake. These factors accounted for 51% of the superiority of behavioral counseling over nutrition education. PMID- 15546226 TI - Relationship of social support and social burden to repeated breast cancer screening in the women's health initiative. AB - Direct and interactive effects of social support, social burden (caregiving, negative life events, and social strain), education, and income on repeated use of breast cancer screening among a large (N=55,278), national sample of postmenopausal women participating in the Women's Health Initiative observational study were examined. Repeated screening decreased as emotional/informational support and positive social interactions decreased (ps<.01). Repeated mammography decreased with frequent caregiving (p<.01). Less social strain reduced the frequency of repeated breast self-examinations (BSEs; ps<.01), but frequent caregiving and more negative life events increased repeated use of BSE (ps<.01). Interactive effects suggested that emotional/informational but not tangible support is associated with repeated mammography and clinical breast examinations (ps<.01) and may be particularly important among low-income older women, especially those burdened by caregiving. PMID- 15546227 TI - Finding benefit in breast cancer during the year after diagnosis predicts better adjustment 5 to 8 years after diagnosis. AB - Cancer patients experience positive as well as adverse consequences from diagnosis and treatment. The study reported here examined longer term reverberations of such experiences. A set of benefit-finding items along with measures of well-being were completed by 230 early-stage breast cancer patients in the year postsurgery. Four to 7 years later, 96 of them again completed measures of well-being. Controlling for initial distress and depression, initial benefit finding in this sample predicted lower distress and depression at follow up. PMID- 15546228 TI - Is it beneficial to involve a family member? A meta-analysis of psychosocial interventions for chronic illness. AB - Links between chronic illness and family relationships have led to psychosocial interventions targeted at the patient's closest family member or both patient and family member. The authors conducted a meta-analytic review of randomized studies comparing these interventions with usual medical care (k=70), focusing on patient outcomes (depression, anxiety, relationship satisfaction, disability, and mortality) and family member outcomes (depression, anxiety, relationship satisfaction, and caregiving burden). Among patients, interventions had positive effects on depression when the spouse was included and, in some cases, on mortality. Among family members, positive effects were found for caregiving burden, depression, and anxiety; these effects were strongest for nondementing illnesses and for interventions that targeted only the family member and that addressed relationship issues. Although statistically significant aggregate effects were found, they were generally small in magnitude. These findings provide guidance in developing future interventions in this area. PMID- 15546229 TI - Latent growth curve analyses of peer and parent influences on smoking progression among early adolescents. AB - Social influences on smoking uptake were examined in latent growth curve analyses of data from 1,320 youths assessed 5 times during 6th to 9th grade. Initial smoking stage predicted increases in number of friends who smoked, indicating selection; however, initial number of friends who smoked did not predict smoking stage progression, indicating no significant effect of socialization. Associations over time among smoking stage progression, affiliation with friends who smoke, and parenting behaviors were significant, suggesting dynamic, reciprocal relationships. Parental involvement, monitoring, and expectations provided direct protective effects against smoking progression as well as indirect effects, by limiting increases in number of friends who smoke. These results are consistent with the peer selection hypothesis, confirm the powerful association over time of social influences with smoking, and provide the first evidence that parenting behavior may protect against smoking progression by limiting increases in number of friends who smoke. PMID- 15546230 TI - Psychological response to test results in an ovarian cancer screening program: a prospective, longitudinal study. AB - To identify the psychological impact of receipt of an abnormal yet benign screening test result, the authors examined the response to a transvaginal ultrasound screening (TVS) test for ovarian cancer (OC) in asymptomatic women (N=540) undergoing an initial TVS screening test. Interviews were conducted prior to undergoing TVS screening and at 2 weeks and 4 months following this baseline. Women receiving an abnormal yet benign TVS test result (n=33) reported elevated OC-specific, but not general, distress at 2-week follow-up. Distress returned to baseline levels at 4-month follow-up. Consistent with the monitoring process and cognitive-social health information processing models, response to an abnormal TVS result was moderated by a monitoring coping style, low optimism, and a family history of OC. PMID- 15546231 TI - Hostility and distraction have differential influences on cardiovascular recovery from anger recall in women. AB - This study investigated the relation of dispositional hostility to cardiovascular reactivity during an anger-recall task and of hostility and distraction to posttask recovery in 80 healthy women (ages 18-30). Half were randomly assigned to distraction during recovery. Hostility predicted slower systolic blood pressure and preejection period during recovery. Distraction was related to faster cardiac recovery, higher high-frequency (HF) power, lower low-frequency (LF) power and LF:HF ratios, and lower state anger and rumination during recovery. These results indicate deleterious influences of hostility on cardiovascular recovery but not during anger recall. The findings also show beneficial effects of distraction in expediting cardiovascular recovery, possibly through reducing rumination and anger. PMID- 15546232 TI - Relationships among health factors and everyday problem solving in african americans. AB - This study examined whether measures of health status enhance the prediction of performance on everyday problem solving in adult African Americans. The sample consisted of 209 community-dwelling African American adults with a mean age of 66.82 years (SD=7.95). The following variables were included in the analysis: Everyday Problems Test (EPT), summary index of chronic illnesses (cardiovascular disease, hypertension, arthritis, stroke, and diabetes), self-rated health (current health, health in the past month, health compared with others, health compared with 5 years ago), and demographic information. Using hierarchical regression and follow-up communality analysis, the authors found that the number of chronic illnesses and self-rated health as compared with 5 years prior were significant and unique predictors of performance on the EPT but did not account for all of the demographic-related variance. The results indicate that health indices contribute to the variability in everyday cognition in this understudied population. PMID- 15546233 TI - Repression and high anxiety are associated with aberrant diurnal cortisol rhythms in women with metastatic breast cancer. AB - Previous research has provided evidence of autonomic, endocrine, and immunological dysregulation in repressers and a possible association with cancer incidence and progression. Recently published data from the authors' laboratory demonstrated that flatter diurnal cortisol slopes were a risk factor for early mortality in women with metastatic breast cancer. In the current analysis of this same sample (N=91), the authors tested differences at baseline between groups scored using the Weinberger Adjustment Inventory on diurnal cortisol slope and mean cortisol levels. When compared with self-assured and nonextreme groups, the represser and high-anxious groups had a significantly flatter diurnal slope. Diurnal slope was similar for repressers and high-anxious groups. Groups did not differ on mean cortisol levels, nor did they differ on intercept (morning) values. PMID- 15546234 TI - Gender and the natural history of self-rated health: a 59-year longitudinal study. AB - Self-ratings of health are uniquely predictive of morbidity and mortality, and they encompass people's evaluations of many medical, psychological, and social conditions in their lives. However, the longitudinal trajectory of self-rated health has not been evaluated to date. In the present study, 59-year longitudinal multilevel analyses (1940-1999) of data from 1,411 men and women revealed that self-rated health was relatively stable until age 50 and then began to decrease in an accelerating fashion through the rest of the life course. Men had higher self-rated health throughout most of adulthood than did women but had steeper linear rates of decline. As a result, the gender difference in self-rated health disappeared by late adulthood. PMID- 15546236 TI - Cervical cancer in the asian pacific-epidemiology, screening and treatment. AB - Squamous cell carcinoma (SCC) of the cervix continues to be a major problem in many areas of the Asian-Pacific, particularly in the Indian subcontinent and Papua New Guinea, and to a lesser extent in South-East Asia, Korea and Mongolia. In contrast, levels in the developed countries of the region are low, as is also the case for the Muslim countries of Western Asia, and mainland China. Incidence generally mirrora associated mortality, although with some exceptions reflecting facilities and infrastructure for early detection. Over the last 25 years there has been a marked decrease in incidence rates across most of the Asian Pacific, although less pronounced in India than elsewhere, and there are exceptions where the incidence is on the increase. The predominant risk factor is well established to be persistent infection with a high risk 'oncogenic' type of human papilloma virus (HPV), along with multiple partners, other sexually transmitted diseases and smoking. Consumption of vegetables, in contrast, appears to be protective. Hormonal factors may also play some role. Modifying factors may either impact on neoplasia by directly influencing the processes underlying carcinogenesis, or indirectly by affecting persistence of viral infections. For primary prevention, avoidance of repeated infections and smoking, as well as a high antioxidant intake may be beneficial. Vaccines against HPV also have promise for the future, but a better understanding of the mechanisms underlying spontaneous clearance of both infection and cervical intraepithelial neoplasia (CIN) of different grades is also essential for optimal intervention. For screening, the choice of whether the PAP smear, HPV testing or some form of visual inspection are utilized depends on the resources which are available, all approaches having their own advantages and disadvantages, but with similar sensitivity and specificity. One complication is the increase in adenocarcinoma of the cervix which has been reported in some countries, for which risk factors and most effective screening may differ from the SCC case. A focus on high risk groups like sex workers might be warranted where financial and technical support are limited. If cervical intraepithelial neoplasias are detected then cryotherapy or the loop electrosurgical excision procedure (LEEP) are effective for their removal. Control of cancer of the cervix, however, demands that a comprehensive approach to screening and management is adopted, necessitating major training of personnel and provision of appropriate resources. PMID- 15546237 TI - Allelic variation of GSTT1, GSTM1 and GSTP1 genes in North Indian population. AB - Glutathione S-transferase (GST) enzymes are involved in detoxification of many potentially carcinogenic compounds. Homozygous deletions or null genotypes of GSTT1 and GSTM1 genes and an A to G substitution at nucleotide 313 in GSTP1 have been reported in different populations. Intra-ethnic as well as interethnic differences are known to exist in the frequencies of the above GST genes. The present study was therefore undertaken to determine the prevalence of GSTM1 and GSTT1null alleles, as well as the GSTP1 gene polymorphism, in 370 healthy individuals in a North Indian population. Genotyping of M1 and T1 was performed using a multiplex polymerase chain reaction and the GSTP1 polymorphism was determined by the polymerase chain reaction/restriction fragment length polymorphism (PCR-RFLP) method. The frequencies of GSTM1 and GSTT1 null alleles in normal healthy individuals were observed to be 33.0% and 18.4% respectively. In 7.0% of individuals' concomitant lack of M1 and T1 genes were observed. For GSTP1, wild (Ile/Ile), heterozygous (Ile/Val) and mutant (Val/Val) genotypes were observed for 44.3%, 50.3% and 5.4% of individuals respectively. The prevalence of the M1 null allele is significantly lower than those documented for English, Turkish, Chinese, Caucasians, Japanese and white (Brazilian and American) populations. However, a significantly higher frequency for T1 null was reported in Chinese and Japanese population. Furthermore, Japanese and African American populations have exhibited significantly higher frequencies of wild and mutant P1 genotypes, respectively, than the Indian population. Thus, our results signify an impact of ethnicity and provide a basis for future epidemiological and clinical studies. PMID- 15546238 TI - Knowledge of the effects of sun exposure of Turkish high school students and their sun bathing habits. AB - Cancer, long a serious problem in developed countries, is now becoming a serious health concern throughout the world. There has been an alarming increase in the number of new cases of melanoma each year, this cancer increasing at a faster rate than any other neoplasm in some regions. This may be a result of depletion of the ozone layer. An association between non-melanocytic skin cancer and exposure to the sun appears to have first been suggested in 1894; it was not until about 1952 that it was argued that exposure to the sun also causes melanoma. It is commonly believed that skin cancers develop only after long-term exposure to UVR. At the individual level, people who live in sunnier climates comparatively have a higher risk for skin cancer than do people who live in colder climates. This is particularly the case for migrant populations with a fair skin. The present study was conducted to evaluate the knowledge of the young generation in Turkey related to the side effects of sun exposure, and their sun bathing habits. Sun sensitivity, use of sunscreens, sunbathing habits and protective behaviours were determined for a total of 1244 high school students from the answers to a questionnaire completed by them. Data from 627 (50.4 %) females and 617 (49.6 %) males were evaluated. All were aged between 15 and 18 years. Sunscreen use was found to be higher in female students (59 %) than in their male counterparts (45.8 %). The wearing of sunglasses was reported by 36.7 %, whereas the incidence of hat wear was found to be 55.1% overall. The results of this study indicate that, although most high school students are aware of the side effects of sun exposure, they do not pay sufficient attention to protective behaviour. A further study should now be conducted to evaluate the use of sunscreens in a large group. PMID- 15546239 TI - An epidemiological study of HBV, HCV and HTLV-I in Sherpas of Nepal. AB - An epidemiological study of hepatitis viruses type B (HBV) and type C (HCV) and human T-cell leukemia virus type I (HTLV-I) was carried out among 103 residents (male:female=61:42) regarded as Sherpas, at Lukla (Solukhumbu district), Nepal in 2004. Blood was drawn from apparently healthy volunteers at ages of 28.8+12.3 (range 15-66) years. HBsAg, HBsAb, HBcAb, and HCV Ab were measured by microparticle enzyme-immunoassay, and HTLV-I Ab was measured by particle agglutination. Prevalence of HBsAg(+), HBsAb(+), HBcAb(+), and HBsAb(+) or HBcAb(+) were 1.9% 22.3%, 24.3%, and 28.2%, respectively. For HCV Ab, only a borderline reaction was observed in one sample, and for HTLV-I Ab all samples were negative. Nucleotide sequencing of the PreS1, PreS2, and S genes revealed that HBV among Sherpas to be of the A' (or Aa) genotype, which is prevalent among Nepalese but rare in native Tibetans, suggesting transmission within Nepal rather than association with ancestors' migration from Tibet as the origin. This is the first report of Himalayan Sherpas' state of infection with HBV, HCV, and HTLV-I. PMID- 15546240 TI - Do Japanese take more folate from traditional Japanese dish than is conventionally estimated?-Actual folate contents in hospital diets and marketed lunch boxes. AB - Folic acid deficiency is very rare in Japan. However, recent concerns in relation to causal relationships between cancer and low folic acid levels necessitate information on actual folate intake by Japanese, which has hitherto not been well studied. We therefore evaluated folate contents of a hospital diet for 7 consecutive days and of 16 lunch boxes purchased at convenience stores. Dietary intake was assessed for weighed food items after cooking using our previously published data on folate contents of various foods and also by using Standard Tables of Food Composition in Japan (STFCJ). Mean daily folate intake from the hospital diet was 723.9 microg/day using our data but only 359.2 microg/day if calculated using the STFCJ. Twenty-nine % of the total daily folate intake was through rice. Mean folate intake through lunch boxes was 491.7 microg/day by our tabulation and 139.5 microg/day with the STFCJ. Folic acid intake of Japanese is far above the levels taken in other countries and over half of this amount is taken from rice. Levels of folic acid contents (especially in rice) listed in the STFCJ are too low and revision is strongly suggested. PMID- 15546241 TI - Hodgkin's disease: assessment of treatment and survival rates in Iran. AB - BACKGROUND AND OBJECTIVE: Each year an estimated of 7,500 new cases of Hodgkin's disease are reported in the United States. It is a type of malignancy, where 75% of patients can recover and be cured with modern therapeutic approaches if presentation is in an early stage. While primary prevention is not a focus of attention, prevention of mortality is thus possible. The main objective of this investigation was to assess the current situation with the disease in Iran, with determination of 5- and 10-year-survival rates. MATERIALS AND METHODS: This retrospective, descriptive, analytical and cross-sectional study was performed on patients identified in Tabriz Shahid Ghazi hospital. The information obtained through medical files was organized and the rate of response to treatment and overall survival (OS) were computed. Resulting data were analyzed using SPSS10 and Chi-square software. RESULTS: Overall, there were 121 male patients (67%) and 59 females (33%). The patient age (with a median of 31.8+/-17.1 years) did not show any effect on survival rate. Neck masses were the most common (40%) complaint among new patients, mostly classified as stage III. Mixed cellularity (47.2%) accounted for the most common histological subtype. Complete remission was achieved with the ABVD chemotherapy regimen, included in 37.6% of overall chemotherapy regimens. The five- and 10-year-survival rates were determined to be 65 and 61.3 percent respectively. CONCLUSION: Chemotherapy was a significantly more effective treatment compared to other modalities, and provided complete remission in 52.7% of patients. Overall, 5- and 10-year-survivals were shown to be highest in patients treated with ABVD and a variant of the MOPP regimen, respectively. As general conclusions, early diagnosis, on time management of the patients, and use of appropriate treatment modalities provide significant prevention of mortality in Hodgkin's disease patients. PMID- 15546242 TI - Co-risk factors for HPV infection in Northeastern Thai women with cervical carcinoma. AB - HPV infection is the main cause of cervical cancer; however, factors that promote and maintain HPV infection are still unclear. This study was designed to search for factors responsible for the HPV infection in Northeastern Thai women. A total of 190 volunteers with a normal histopathologic appearance of cervix as controls (n=100) and with squamous cell cervical carcinoma (SCCA) (n=90) were the subjects. Variables of risk factors including sexual behaviors, history of reproduction, history of sexually transmitted diseases and smoking were conducted with self-report and direct interview. Number of sexual partners and smoking history increased the likelihood of high-risk HPV infection. Multiple sexual partners showed significantly higher 3.94-fold risk for HPV infection (95% CI = 1.82-8.82, p-value<0.001). Smoking history of partner increased the risk for HPV infection 3.03-fold (95%CI=1.42-6.58, p-value< 0.002). After OR were adjusted, significant difference was still observed in the number of sexual partners (p value <0.0001) and smoking history of the partner (p-value<0.005). To decrease the incidence of cervical cancer, we should prevent HPV dissemination and be on the alert for having multiple sexual partners and a partner's smoking habit, which must be included in our public health planning. PMID- 15546243 TI - A study of cancer patterns among inpatients of public hospitals in Iran. AB - Cancer is becoming an increasingly important cause of premature mortality in developing countries as their populations expand and their lifestyle becomes westernized. The aim of this study was to determine the frequency distribution of various neoplasms among Iranian inpatients, their demographic status and length of stay involved in hospitals during 2000-2002. Records of 17447 inpatients who were hospitalized with malignant tumors in Iranian public hospitals during 2000 2002 were studied. The neoplasms had been coded and classified according to International Classification of Diseases, 10th Revision (ICD-10). The frequency distribution of cancer patients was evaluated by age, sex, place of residence and the length of stay at hospital. There were 9470 male patients (54.3%) and 7977 females (45.7%). The mean +/- sd age was 51.2+/-20.6 with a median of 55 years. The average for females (49.4+/-19.2 yrs) was significantly lower than that for males (52.8+/-21.7 yrs) (p<0.001). The five most common cancer sites were the digestive organs (27.6%) followed by lymphoid and haematopoitic tissues (21.4%), breast (10.2%), respiratory and intrathoracic organs (8.8%) and skin (6%). These accounted for 74% of all malignancies. Some 31% of women's cancers were found in breast or genital organs compared to only 7.6% for males. The male/female ratio was 1.19 with the highest being 2.85 for respiratory organs and the lowest being 0.04 for breast. Mean age of male patients with cancer of the digestive organs, respiratory and intrathoracic organs and skin was significantly lower than that of females (p<0.001). The median length of stay of patients in hospitals each time they were admitted was 6 days. Of the cancer patients, 77.7% lived in urban areas which include only 64.5% of the population. Rural patients with skin cancer accounted for 32.1%. In conclusion, the results of this study carry important implications for future health planning strategies and provide a baseline for further studies on the evaluation of malignancies in Iran. PMID- 15546244 TI - A case-control study of colorectal cancer in relation to lifestyle factors and genetic polymorphisms: design and conduct of the Fukuoka colorectal cancer study. AB - A case-control study was designed to elucidate roles of dietary and other behavioral influences, in combination with genetic susceptibility factors (genetic polymorphisms), in colorectal carcinogenesis. Both cases and controls were residents in Fukuoka City and three adjacent areas. Cases were patients undergoing surgery for a first diagnosis of colorectal cancer at 8 hospitals in the study area, and controls were randomly selected in the community by frequency matching with respect to the expected distribution by sex, age (10-year class), and residence. Dietary and other lifestyle factors were ascertained by in-person interview, and venous blood was obtained for genotyping and possible biochemical measurements. The cases were interviewed at each hospital during the period from 2000 to 2003, and controls were surveyed during the period from 2001 to 2002. A total of 840 cases of colorectal cancer and 833 controls were interviewed with participation rates of 80% for the cases and 60% for the controls. Informed consent to genotyping was obtained from 685 cases and 778 controls. Further details of the design and conduct are described with respect to methodological aspects. PMID- 15546245 TI - Cumulative risk and trends in prostate cancer incidence in Mumbai, India. AB - BACKGROUND: Information relating to cancer incidence trends in a community forms the scientific basis for the planning and organization of prevention, diagnosis and treatment of cancer. We here estimated the cumulative risk and trends in incidence of prostate cancer in Mumbai, India, using data collected by the Bombay Population-based Cancer Registry from the year 1986 to 2000. METHODS: During the 15 year period, a total of 2864 prostate cancer cases (4.7% of all male cancers and 2.4% of all cancers) were registered by the Bombay Population-based Cancer Registry. For evaluation of the trend, we applied a linear regression model based on the logarithm of the observed incidence rates. The annual percentage changes were also computed for the evaluation. Cumulative incidence rates percentages were calculated by adding up the age specific incidence rates at single ages and then expressed as a percentage. RESULTS: Analysis of the trends in age-adjusted incidence rates of prostate cancer during the period 1986 to 2000 showed no statistically significant increase or decrease and the rates proved stable across the various age groups (00-49, 50-69 and 70+) also. The probability estimates indicated that one out of every 59 men will contract a prostate cancer at some time in his whole life and 99% of the chance is after he reaches the age of 50. CONCLUSION: The stability in age adjusted-incidence rates indicates that there are no changes in the etiological factors for prostate cancer in Mumbai, India. These findings may be of general interest because changes in diagnostic practices are confounded in the time trends of prostate cancer change in many western countries preventing inferences on the changes in risk. PMID- 15546246 TI - Prostate specific antigen for screening for prostate cancer: an appraisal of Thai reports. AB - Prostate cancer is the most common cancer in male populations in many parts of the world. It is a slowing growing deadly cancer with very few signs and symptoms in the early stage. For screening, prostate specific antigen (PSA) has been proposed as a marker in the serum. It is widely employed in western countries, but use of PSA for prostate cancer screening in developing Asian countries is not generalized. Here, the author performed an appraisal on the diagnostic properties of serum PSA in screening prostate cancer among the Thais. Four reports from the literature were recruited for further metanalysis of a total of 1,321 cases. The overall diagnostic activity with regard to sensitivity, specificity, false positive and false negative rates, values being 95.8 %, 66.2 %, 33.8 % and 4.2 %, respectively. Therefore the test has good sensitivity, and sufficiently good diagnostic properties for screening when compared to digital rectal examination (DRE). However, serum PSA cannot be used for a definitive diagnosis, for which pathological confirmation is also necessary. PMID- 15546247 TI - A pilot study on genotype announcement to induce smoking cessation by Japanese smokers. AB - BACKGROUND: Genotype announcements related to susceptibility to hazardous effects of smoking may be effective to induce smoking cessation. METHODS: Subjects were municipal government employees, 63 young smokers employed in the previous year and 59 smokers with more than 45 pack-years, who were invited to educational sessions against smoking held in December 2003 and February 2004, respectively. In the session, those who wished genetic susceptibility tests (GSTM1, GSTT1, and NQO1 C609T) were enrolled in the study. The smoking habit was ascertained three times: at the session, one month later, just before the genotype announcement, and at the follow-up three months after the announcement. RESULTS: Fifty eight (92.1%) and 49 (83.1%) smokers participated in the study, respectively. One out of 58 smokers was not a habitual smoker, so was not included in the analysis. The smoking cessation rates were 15.8% (9 participants) and 6.1% (3 participants) just before the genotype announcement, and 7.0% (4 participants) and 10.2% (5 participants) at the follow-up, respectively. All subjects were satisfied with the genotype testing except for two who rather regretted participating, but one of whom actually quit smoking. CONCLUSION: The present pilot study without controls indicated that the effects of genotype announcements in this framework on smoking cessation were less than might have been expected. The temporary effect of the session on younger smokers may have been due to the participation per se. The potential effects of genotype announcements for heavy smokers should now be examined in studies with adequate controls. PMID- 15546248 TI - Mortality report of malignant tumors in SheXian, Hebei Province, China, from the 1970's to the present. AB - BACKGROUND: Investigations into mortality from malignant tumors were initiated in the 1970's in Hebei Province, China, and especially for esophageal cancer the rates were high, Shexian county ranking in first place of the towns that were surveyed. METHODS: Since the 1970's, a register system for all causes of death has been in place. Data for the decades of the 1970's, 1980's, 1990's and 2000's century were here checked and analyzed by SPSS software. RESULT: From the decades of the 1970's onward, the mortality rates of malignant tumors/100,000 were 272.0, 260.1, 211.7 and 180.1, respectively, with significant differences over time (x2 =240.5, P<0.001). The main malignant tumors were esophageal, gastric, liver, lung and cervix cancers. The sum of their percentages of all cancer deaths were 92.1% in the 1970's, 91.6% in the 1980's, 92.1% in the 1990's and 93.9% in the 21st century. The sex ratios (male vs female) were 1.5, 1.5, 1.7 and 2.0 respectively, with an ascending trend. Mortality rates of malignant tumors increased with age, with an obvious geographic distribution. The highest mortality of malignant tumors was evident in the area where the Qingzhang and Zhuozhang rivers join. CONCLUSION: From 1970's to the beginning of the 21st century, the mortality rate of malignant tumors has shown a declining trend. The main responsible cancers are in the esophagus, stomach, liver, and lung. Through great efforts for prevention, obvious decrease for esophageal cancer and cervix cancer has been achieved, but the mortality rate for gastric cancer remains high. PMID- 15546249 TI - Comparison of lifestyle risk factors by family history for gastric, breast, lung and colorectal cancer. AB - To assess the theoretical impact of lifestyle of a cancer family history in first degree relatives (CFH) and clarify interactions between CFH and lifestyle factors, hospital-based comparison and case-reference studies were conducted in Nagoya, Japan. Totals of 1988 gastric, 2455 breast, 1398 lung and 1352 colorectal cancer patients, as well as 50,706 non-cancer outpatients collected from 1988 to 1998, were checked for lifestyle factors, which included dietary and physical exercise habits, as well as smoking/drinking status. General lifestyle factors with non-cancer outpatients did not differ by the CFH status. Case-reference analyses showed that frequent intake of fruits, raw vegetables, carrots, pumpkin, cabbage and lettuce, as well as frequent physical exercise, were associated with decreased risk for all four sites of cancer, while habitual smoking increasing the risk of gastric, and more particularly, lung cancer. Interestingly, the study revealed the magnitude of odds ratios for the above lifestyle factors obtained from CFH positives to be similar to those from CFH negatives for these four sites of cancer. There were no significant interactions between CFH and any particular lifestyle factor. In conclusion, our results suggest no appreciable influence of CFH on lifestyle related risk factors for gastric, breast, lung, and colorectal cancer. Habitual smoking increased, while frequent physical exercise and raw vegetables intake decreased cancer risk, regardless of the CFH status. PMID- 15546250 TI - "I don't really know, so it's a guess": women's reasons for breast cancer risk estimation. AB - Women of all ages have been found to overestimate both the incidence and the mortality rate from breast cancer and the reasons for this are unclear. A qualitative study asked eighty three women (mean age = 44 years) how likely they thought they were to get breast cancer and to explain the reasoning behind their choice. Based on their responses, women's perceptions were categorised as: no risk (5%); reasonably accurate (30%); overestimated (22%); and greatly overestimated (43%). Four main themes emerged from the reasons given: 'Don't know/guess', 'family history' of breast cancer,'age' related reasoning, and making their decision from the information sheet read prior to answering the questions. The information currently available to women may be creating falsely high estimates of their risk of developing breast cancer as the risk factors of age and family history appear to be poorly understood. Meaningful communication of health risk in need of further improvement if it is to be useful in changing health related knowledge and behaviours. PMID- 15546251 TI - Inhibitory effects of crude alpha-mangostin, a xanthone derivative, on two different categories of colon preneoplastic lesions induced by 1, 2 dimethylhydrazine in the rat. AB - The purpose of this study was to examine whether crude alpha-mangostin (a major xanthone derivative in mangosteen pericarp (Garcinia mangostana)) has short-term chemopreventive effects on putative preneoplastic lesions involved in rat colon carcinogenesis. The crude preparation was obtained by simple recrystallization of an ethylacetate extract of mangosteen pericarps. A total of 33 five-week-old male F344 rats were randomly divided into 5 experimental groups. Rats in groups 1-3 were given a subcutaneous injection of 1,2-dimethylhydrazine (DMH)(40 mg/kg body weight) once a week for 2 weeks. Starting one week before the first injection of DMH, rats in groups 2 and 3 were fed a diet containing 0.02% and 0.05% crude alpha-mangostin, respectively, for 5 weeks. Rats in group 4 also received the diet containing 0.05% crude alpha-mangostin, while rats in group 5 served as untreated controls. The experiment was terminated 5 weeks after the start. Dietary administration of crude alpha-mangostin at both doses significantly inhibited the induction and/or development of aberrant crypt foci (ACF) (P<0.05 for 0.02% crude alpha-mangostin, P<0.01 for 0.05% crude alpha-mangostin), when compared to the DMH-treated group (group 1). Moreover, treatment of rats with 0.05% crude alpha-mangostin significantly decreased dysplastic foci (DF) (P<0.05) and beta-catenin accumulated crypts (BCAC) (P<0.05), to below the group 1 values. The proliferating cell nuclear antigen (PCNA) labeling indices of colon epithelium and focal lesions in groups 2 and 3 were also significantly lower than in group 1 and this effect occurred in a dose dependent manner of the crude alpha mangostin. This finding that crude alpha-mangostin has potent chemopreventive effects in our short-term colon carcinogenesis bioassay system suggests that longer exposure might result in suppression of tumor development. PMID- 15546252 TI - A survey on breast self-examination among Thai adult women in a rural distant subdistrict. PMID- 15546253 TI - Thymalfasin: an immune system enhancer for the treatment of liver disease. AB - Thymalfasin (thymosin-alpha 1) is an immunomodulating agent able to enhance the Th1 immune response. It has been evaluated for its immunomodulatory activities and related therapeutic potential in several diseases, including chronic hepatitis B and C, AIDS, primary immunodeficiency diseases, depressed response to vaccination and cancer. The basis for effectiveness in these conditions is primarily through modulation of immunological responsiveness, as thymalfasin has been shown to have beneficial effects on numerous immune system parameters and to increase T-cell differentiation and maturation. Thymalfasin is responsible for reconstitution of immune function when thymic tissue is given back to thymectomized animals. In addition, thymalfasin has been shown to have efficacy in multiple experimental models of immune dysfunction, mainly, infectious diseases such as hepatitis (woodchuck) and influenza (mouse), and cancer such as melanoma (mouse) and colorectal carcinoma (rat) where thymalfasin has shown antitumor effects. PMID- 15546254 TI - Thymalfasin (thymosin-alpha 1) therapy in patients with chronic hepatitis B. AB - Chronic hepatitis B virus (HBV) infection is a serious clinical problem because of its worldwide distribution and potential adverse sequelae. Globally, there are approximately 350 million people infected with chronic HBV, 75% of whom live in the Asia-Pacific region. Interferon-alfa and direct antiviral agents such as lamivudine and adefovir are effective in the therapy of chronic HBV infection but the efficacy is far from satisfactory, particularly in perinatally infected patients, patients with lower ALT levels and those with HBeAg-negative chronic hepatitis B. Thymalfasin (thymosin-alpha1) is an immunoregulatory agent able to enhance Th1 response. It has been shown to trigger maturational events in lymphocytes, to augment T-cell function, and to promote reconstitution of immune defects. Studies are underway in both monotherapy and combination therapy with thymalfasin and interferon and results are promising. PMID- 15546255 TI - Combination therapy of thymalfasin (thymosin-alpha 1) and peginterferon alfa-2a in patients with chronic hepatitis C virus infection who are non-responders to standard treatment. AB - The worldwide spread of hepatitis C virus is enormous; chronic hepatitis C virus infection is a leading cause of liver cirrhosis and hepatocellular carcinoma. While treatment options have improved substantially over the last decade, responses are still disappointing, particularly in certain difficult-to-treat groups such as patients who are immunosuppressed or have decompensated disease. Preliminary studies have indicated that combined treatment strategies may provide effective approaches for the future. The combination of thymalfasin with pegylated interferon is currently a promising option for the treatment of patients with chronic hepatitis C virus infection. An ongoing phase 3 study in the USA should provide much needed data to improve the outcome for these patients. PMID- 15546256 TI - Triple combination of thymalfasin, peginterferon alfa-2a and ribavirin in patients with chronic hepatitis C who have failed prior interferon and ribavirin treatment: 24-week interim results of a pilot study. AB - Despite steady progress in antiviral treatment for patients with chronic hepatitis C virus (HCV), many patients still have detectable serum HCV RNA levels by the end of interferon-based treatment and are known as virological non responders. Re-treatment of these patients not responding to previous therapy remains challenging. Studies of the dynamics of the HCV population show a marked decline in new cases since 1996; however, the relative proportion of non responders is expected to increase over time and, similarly, the number of patients eligible for first-line treatment is expected to decrease. The current standard of care for treatment involves the use of pegylated interferons in combination with ribavirin. However, many difficult-to-treat groups still have low response rates. Newer combinations are being investigated to optimize chances of attaining a sustained response in these groups: one such triple therapy regimen is peginterferon alfa-2a, ribavirin and thymalfasin, which was given to 23 previously non-responder patients. Viral response was 60.8% at week 12 and 47.8% at week 24. These preliminary results encourage further evaluation of this promising combination. PMID- 15546257 TI - Topographic and dipolar analysis of laser-evoked potentials during migraine attack. AB - OBJECTIVE: The aim of this study was to perform further evaluation of laser evoked potentials (LEPs) during migraine attacks using multichannel recording and topographic analysis. Specifically, this study aimed to confirm the pattern previously observed in acute migraine, while also defining the components of LEPs that are mainly modified during headache, as well as the correlation between features of LEPs and clinical variables. In addition, we aimed to conduct a dipolar source analysis of the main LEP waves in migraine patients to check the variability in the source location of LEPs during acute migraine. BACKGROUND: An amplitude enhancement of LEPs was previously detected during migraine attack using a single scalp derivation on the vertex; hyperalgesia to heat stimuli was also detected for both the face and hand. METHODS: Eighteen patients suffering from migraine without aura were analyzed. The supraorbital zones and the dorsum of the hand were stimulated on both the symptomatic and nonsymptomatic sides in all patients. The LEPs were recorded via 25 scalp electrodes. Dipolar source analysis of the P2 components was performed using a spherical model in all patients and using a realistic Magnetic Resonance model in four patients. RESULTS: During attacks, the later waves, and particularly the P2 component, were significantly enhanced; the amplitude of the P2 component obtained during the attack by stimulation of the supraorbital zone on the side of the headache was significantly correlated with the intensity of pain and the frequency of headache. In our patients, the P2 wave was generated in the anterior cingulate cortex, with a shift toward its rostrocaudal portion, and was mainly devoted to elaboration of the emotive compound of pain during migraine attack. CONCLUSIONS: Cortical activation by laser stimuli during migraine attack was confirmed. This effect was more pronounced in patients with a higher frequency of migraine attacks. This may be due to a lack of inhibitory control over the transmission of pain to the cortex. The increased activation of cortical areas devoted to attention and emotion may be linked to headache. PMID- 15546258 TI - Chemokine levels in the jugular venous blood of migraine without aura patients during attacks. AB - OBJECTIVE: To investigate changes in the levels of calcitonin gene-related peptide and its intracellular messenger cyclic adenosine monophosphate in serial samples of internal jugular blood taken from migraine patients without aura assessed during attacks, and to assess their relationship with the levels of IL 8, MCP-1, and RANTES in the same samples. BACKGROUND: Calcitonin gene-related peptide, the marker of trigeminovascular activation, is released in both the internal and external jugular venous blood of migraine patients during attacks. Experimental evidence demonstrated that when released from C-type sensory neurons in inflammatory pain models, it differentially induced expression of neutrophil chemotactic chemokine IL-8, but not monocyte chemotactic chemokine MCP-1 or lymphocyte chemotactic chemokine RANTES. These chemokines were never investigated in migraine. DESIGN/METHODS: Eight migraine without aura patients were admitted to the hospital during the attacks. Internal jugular venous blood samples were taken immediately after catheter insertion, at the 1st, 2nd, and 4th hours after attack onset, and within 2 hours from its cessation. The levels of the sensory neuropeptide calcitonin gene-related peptide and the messenger cyclic adenosine monophosphate were measured by RIA method, and those of IL-8, MCP-1, and RANTES were measured by ELISA method. RESULTS: Higher calcitonin gene-related peptide levels were found in the internal jugular venous blood of migraine without aura patients compared with the time of catheter insertion (ANOVA: P<.0001) with a peak at the first hour (52.6+/-9.2 ng/mL). A transient increase in IL-8 was observed at the 2nd and 4th hours (P<.01 and P<.002, respectively), whereas no changes in the levels of MCP-1 and RANTES were found at any time of the study. The increase in IL-8 was accompanied by a parallel increase in cyclic adenosine monophosphate. CONCLUSIONS: The present study confirms previous findings of an increase in calcitonin gene-related peptide in internal jugular venous blood of migraine without aura patients during attacks. The transient increase in the levels of IL-8 concurs with the results of recent experimental research showing a calcitonin gene-related peptide-induced activation of IL-8 gene expression, but not RANTES and MCP-1, via the transcriptional factor AP-2, which mediates transduction in response to cyclic adenosine monophosphate. Although IL-8 is transiently increased during migraine attacks, an accumulation of leukocytes secondary to neurogenic inflammation is unlikely, as it is for other inflammatory events, because they are self limiting. Other events, including nitric oxide production, may contribute to counteract meningeal transvascular leukocyte migration during migraine attacks, as suggested by the model of sterile inflammation. PMID- 15546259 TI - Long-term tolerability of sumatriptan nasal spray in adolescent patients with migraine. AB - OBJECTIVE: This 1-year, open-label, multicenter study was designed to assess the long-term tolerability and efficacy of sumatriptan nasal spray 20 mg in adolescent patients with migraine. METHODS: A prospective, multicenter, open label study was conducted in patients aged 12 to 17 years who were allowed to treat an unlimited number of migraines at severe, moderate, or mild pain intensity with sumatriptan nasal spray for up to 1 year. All patients started the study at the 20-mg dose of sumatriptan nasal spray. Dose could be adjusted downward to 5 mg at the discretion of the investigator to optimize therapy. RESULTS: A total of 484 adolescent migraineurs treated 4676 migraines with sumatriptan nasal spray 20 mg (3593 during the first 6 months and 1083 during the second 6 months). A total of 3940 migraines and 699 migraines were treated with one and two 20-mg doses of sumatriptan nasal spray, respectively. Only 10 patients (treating 42 migraines) took the 5-mg dose of sumatriptan nasal spray. The overall percentage of migraines treated with either one 20-mg dose or one, two, or three 20-mg doses with at least 1 drug-related adverse event was 19%. The most common specific drug-related adverse event was unpleasant taste, reported in 17% of migraines. No other single drug-related adverse event was reported in more than 1% of migraines over the 1-year treatment period. When unpleasant taste was excluded from the adverse-event tabulations, the percentages of migraines with at least 1 drug-related adverse event after one or one, two, or three 20-mg doses declined to 4% and 3%, respectively. No patient experienced any drug-related changes in 12-lead ECGs, vital signs, or nasal assessments; and no clinically meaningful changes in clinical laboratory values were observed. Across all migraines with evaluable efficacy data (n=4334), headache relief was reported in 43% of migraines at 1 hour and in 59% at 2 hours after dosing with sumatriptan nasal spray 20 mg. Of the 2561 migraines with headache relief 2 hours postdose, headache recurrence was reported within 24 hours of initial dosing in 7% of migraines. None of the efficacy or tolerability results varied as a function of time in the study (ie, first 6 months vs. second 6 months). CONCLUSION: Sumatriptan nasal spray 20 mg is generally well tolerated and may be beneficial during long-term use by adolescent migraineurs ages 12 to 17 years. PMID- 15546260 TI - Assessment of adverse events associated with triptans--methods of assessment influence the results. AB - BACKGROUND: A recent study conducted in triptan-naive migraine patients showed that tolerability was the second most important attribute of an acute treatment. However, the proportion of patients reporting side effects after any acute treatment may vary with regard to the method of assessment. OBJECTIVES: To contrast two methods of assessing adverse events (prompted and unprompted) in those with headache using triptans. METHODS: This study was conducted in two sites, a headache center in the United States, and a neurology office focusing on headache in Italy. We prospectively surveyed 415 adults with headache, who had been using the same triptan for at least 3 months. Participants were asked about their headache and treatment history. Subjects then completed a standardized questionnaire, assessing adverse events in two different ways. First, subjects were asked if they had any adverse events when using the triptan. After returning the first part of the questionnaire, subjects received a second form, where 49 possible adverse events were listed. We contrasted and correlated both sets of answers. RESULTS: Most patients (U.S.=74.9%, Italy=65.5%) reported no side effects in the unprompted questionnaire. However, most of them (U.S.=62.9%, Italy=54.1%) reported at least one side effect in the prompted questionnaire. Most patients that reported side effects in the unprompted questionnaire said they had just one adverse event, while most reported two or more side effects in the prompted questionnaire. Both in the unprompted and in the prompted questionnaires, most side effects were rated as mild or moderate. Interestingly, 31 (7.5%) subjects (pooling data from both sites together) graded their adverse events as severe in the prompted questionnaire, but had not self-reported them. CONCLUSIONS: (1) When assessing adverse events, the method of data collection may dramatically influence the results. (2) From those subjects who did not self report adverse events after using a triptan, most of them will report positively if presented with a list of side effects. PMID- 15546261 TI - Somatic symptoms in headache patients: the influence of headache diagnosis, frequency, and comorbidity. AB - BACKGROUND: Mood disorders of anxiety and depression are well known to be comorbid with primary headache disorders. Less is known of the comorbidity of other somatic symptoms with headache. METHODS: Headache Clinic patients were screened with the Primary Care Evaluation of Mental Disorders (PRIME-MD), a multidimensional psychiatric screening tool. The prevalence of somatic symptoms was compared by headache diagnosis, frequency of severe headache, and psychiatric diagnosis. Follow-up data were obtained 6 months after consultation. RESULTS: Clinical diagnoses and PRIME-MD data were available for 289 patients. Associated somatic symptoms were more frequent in patients with chronic migraine (mean 5.5, P<.001) and chronic daily headache (CDH) (6.3, P=.008) compared to episodic migraine (4.0); in patients with severe headache >2 days per week compared to 2 days per week had significantly higher somatic counts (P=.01). Six month follow-up data were available for 140 patients. Associated symptoms decreased both for patients with and without decrease in severe headache frequency (mean reduction of 1.0, P=.01 and 0.8, P=.003, respectively). CONCLUSION: Associated somatic symptoms are more common in patients with chronic migraine and CDH, with more frequent severe headaches, and with associated anxiety or depression. Patients with episodic migraine have similar somatic prevalence as a previously studied primary care population. The spectrum of headache disorders may be characterized as showing increasing somatic prevalence as headaches, particularly severe headaches, become more frequent. PMID- 15546262 TI - Genomic abnormalities in patients with migraine and chronic migraine: preliminary blood gene expression suggests platelet abnormalities. AB - BACKGROUND: Migraine has strong genetic and environmental components and may also be a significant contributor to chronic migraine (CM). It is hypothesized that gene expression changes in peripheral blood cells can be used to detect the interaction of these influences. OBJECTIVE: Distinct genomic expression patterns for migraine and CM will be present. These genomic profiles will help clarify the interactions of inheritance and environment. This initial study begins to examine the feasibility of peripheral blood cell genomic analysis to assist in the understanding of the pathophysiology of migraine and CM. METHODS: Blood samples from patients were obtained either during an acute migraine or CM. Genomic expression patterns were analyzed using Affymetrix U95A microarrays. RESULTS: Expression patterns of 7 migraine and 15 CM patients were compared to four distinct control groups (total patients, n=56) including healthy subjects. A group of platelet genes were upregulated in both migraine and CM samples. Different gene expression patterns were also seen between migraine and CM. A group of immediate early genes including c-fos and cox-2 were expressed at higher levels in migraine, whereas specific mitochondrial genes were expressed at higher levels in CM. CONCLUSIONS: Increased expression of platelet genes in patients with migraine and CM suggests similar underlying pathophysiology. The differences seen between migraine and CM in other genes suggest an overlapping but not identical pathophysiology. Further genomic profiling studies will help define these relationships and provide further insights into headache pathogenesis. PMID- 15546263 TI - Serotonin transporter polymorphism and harm avoidance personality in chronic tension-type headache. AB - OBJECTIVE: To investigate the 5-HTT-gene-linked polymorphic region (5-HTTLPR) genotypes and harm avoidance (HA) dimension in chronic tension-type headache (CTH). BACKGROUND: Serotonin transporter protein (5-HTT) is a key modulating protein in synaptic serotonergic neurotransmission. Among serotonin gene-linked polymorphism, promoter located in the regulatory region of the 5-HTTLPR has two alleles (short and long) with different transcriptional efficiencies. The HA personality trait may be heritable and associated with serotonergic neurotransmitter activity. DESIGN: We amplified the 5-HTTLPR by means of polymerase chain reaction and performed genotype polymorphism analyses and we investigated the serotonin-related personality trait by evaluating the HA dimension in tridimensional personality questionnaire (TPQ) in 107 patients with CTH and in 100 healthy controls. RESULTS: We found an excess frequency of the short allele and a different genotype distribution in patients with CTH. S/S genotype frequency was significantly higher in patients with CTH (76%) than in those with controls (59%; P=.02). Patients with CTH had significantly higher HA scores (21.4+/6.3) than controls (16.3+/-6.1). CONCLUSIONS: This suggests a serotonergic activity might be involved in the development of CTH and 5-HTTLPR might be one of the genetically contributing factors. PMID- 15546264 TI - Dynamic mechanical (brush) allodynia in cluster headache. AB - OBJECTIVE: To examine the occurrence of dynamic mechanical (brush) allodynia (BA) in patients with cluster headache (CH). BACKGROUND: Cutaneous allodynia was described in migraine. It was related to sensitization of neurons in the trigeminal nucleus caudalis (TNC). This phenomenon has not been previously described in cluster headache. METHODS: We examined adult patients with episodic or chronic CH for the presence of BA. Demographic data and the characteristics of CH were obtained through a questionnaire. Allodynia testing was performed by repetitively applying a 4x4-inch gauze pad to skin areas in the trigeminal and cervical dermatomes. Degree of allodynia was measured on a 100-mm visual analog scale (VAS). The relations between the location and severity of headache and allodynia were assessed. RESULTS: Ten patients (all male, mean age 39.3) were included in the study. Seven had episodic CH (ECH) and 3 had chronic CH (CCH). Two patients were in acute attack when tested for BA. In total, 4 (40%) of the 10 patients had BA (2 [28.6%] of the 7 with ECH and 2 [66.7%] of the 3 with CCH). Median disease duration was 22 years for patients with BA and 12 years for patients without BA. Of the two patients in acute attack, one had BA, ipsilateral to the headache, which was reduced 20 minutes after treatment, along with reduced headache severity. CONCLUSIONS: This is the first report on the occurrence of cutaneous allodynia in CH. The presence of BA in CH may be related to CH type (episodic vs. chronic) and to the duration of disease. These results support the concept that allodynia in CH may result from a time-dependent process of neuronal sensitization. PMID- 15546265 TI - Individualizing treatment with verapamil for cluster headache patients. AB - BACKGROUND: Verapamil is currently the best available prophylactic drug for patients experiencing cluster headaches (CHs). Published papers usually state 240 to 480 mg taken in three divided doses give good results, ranging from 50% to 80%; others mention higher doses--720, even 1200 mg per day. In clinical practice we found we needed to adapt dosage to individual's time of attacks, in particular giving higher doses before going to bed to suppress severe nocturnal episodes. A few only required 120 mg daily. We therefore evolved a scheme for steady and progressive drug increase until satisfactory control had been achieved. OBJECTIVE: To find the minimum dose of verapamil required to prevent episodic and chronic cluster headaches by supervising each individual and adjusting the dosage accordingly. METHODS: Consecutive patients with episodic or chronic CH (satisfying International Headache Society (IHS) criteria) were started on verapamil 40 mg in the morning, 80 mg early afternoon, and 80 mg before going to bed. Patients kept a diary of all attacks, recording times of onset, duration, and severity. They were advised, verbally and in writing, to add 40 mg verapamil on alternate days, depending on their attack timing: with nocturnal episodes the first increase was the evening dose and next the afternoon one; when attacks occurred on or soon after waking, we advised setting an alarm clock 2 hours before the usual waking time and then taking the medication. Patients were followed-up at weekly intervals until attacks were controlled. They were also reviewed when a cluster period had ended, and advised to continue on the same dose for a further 2 weeks before starting systematic reduction. Chronic cluster patients were reviewed as often as necessary. RESULTS: Seventy consecutive patients, 52 with episodic CH during cluster periods and 18 with chronic CH, were all treated with verapamil as above. Complete relief from headaches was obtained in 49 (94%) of 52 with episodic, and 10 (55%) of 18 with chronic CH; the majority needed 200 to 480 mg, but 9 in the episodic, and 3 in the chronic group, needed 520 to 960 mg for control. Ten, 2 in the episodic and 8 in the chronic group, with incomplete relief, required additional therapy-lithium, sumatriptan, or sodium valproate. One patient withdrew because verapamil made her too tired, another developed Stevens-Johnson syndrome, and the drug was withdrawn. CONCLUSIONS: Providing the dosage for each individual is adequate, preventing CH with verapamil is highly effective, taken three (occasionally with higher doses, four) times a day. In the majority (94%) with episodic CH steady dose increase under supervision, totally suppressed attacks. However in the chronic variety only 55% were completely relieved, 69% men, but only 20% women. In both groups, for those with partial attack suppression, additional prophylactic drugs or acute treatment was necessary. PMID- 15546266 TI - Osmophobia and taste abnormality in migraineurs: a tertiary care study. AB - OBJECTIVES: This study assesses osmophobia and taste abnormality for the first time in a large sample of migraine patients. METHODS: Seven hundred and twenty seven migraineurs were evaluated. Osmophobia, taste abnormality, and perfume or odor trigger were graded from 0 to 3. RESULTS: In patients with data, 24.7% of 673 patients complained of osmophobia (12.5% occasional, 7% frequent, and 5.2% very frequent) and 24.6% of 505 of taste abnormality (13.5% occasional, 6.1% frequent, and 5.0% very frequent). Perfume or odor trigger of acute migraine occurred in 45.5% of 724 patients (22.7% occasional, 10.2% frequent, and 12.6% very frequent). Perfume or odor trigger was associated with osmophobia in 61.5% and taste abnormality 62.1%. Osmophobia without taste abnormality occurred in 28.3% and taste abnormality without osmophobia in 40.3%. A greater percentage of females than males had osmophobia (25.7 vs. 17.5), taste abnormality (25.4 vs. 17.9), and perfume or odor trigger (49.3 vs. 22.1), all P<.0001. CONCLUSIONS: Osmophobia and taste abnormality occur in about one quarter of migraineurs during an acute migraine attack while perfume or odor trigger migraine in almost 50% of patients. Osmophobia and taste abnormality in the acute migraine attack, as well as perfumes or odor as a migraine trigger, are more common in females than in males. PMID- 15546268 TI - Dextroamphetamine pilot crossover trials and n of 1 trials in patients with chronic tension-type and migraine headache. AB - OBJECTIVE: To examine the preventive effects of dextroamphetamine in select small groups of patients with chronic tension-type and migraine headache. BACKGROUND: Neither amphetamine nor methylphenidate is used as a headache preventive. This study was undertaken after a chance observation led one of us to prescribe dextroamphetamine with apparent successes in specific patients with chronic tension-type or migraine headaches. METHODS: Two pilot trials were done. Trial 1 tested patients who were taking dextroamphetamine, while Trial 2 tested patients who had never taken this drug. Each trial obtained full data on eight subjects with chronic tension-type headache and eight subjects with migraine headache. A randomized, double-blinded, controlled, multiple-crossover design was used. Subjects took capsules containing dextroamphetamine or equi-stimulatory caffeine (the control) during four alternating 20-day periods. Trial 1 subjects took their pretrial dextroamphetamine dose at breakfast and lunch. Trial 2 subjects took 10 mg at these times. Subjects recorded the integer from 0 to 3 that represented their headache intensity during the previous 24 hours. The subject's data were the average daily headache grade for the two dextroamphetamine periods and for the two caffeine periods. The differential effect of amphetamine and caffeine on each group of eight subjects and on each individual was analyzed by t-tests. RESULTS: In both trials, the tension-type and migraine groups had lower mean daily headache grades in the amphetamine than in the caffeine periods. P values for these differences indicated that there were real drug effects, on the average, in the migraine groups (P<.05) and suggestive but inconclusive effects in the tension-type groups (P<.10). The individual n of 1 analyses showed that five tension-type and three migraine subjects in Trial 1 and three tension-type and three migraine subjects in Trial 2 had considerably lower mean daily headache grades on amphetamine with P values indicating, at various levels of significance (from P<.05 to P<.001), real amphetamine effects. Twelve of the remaining 18 patients had lower, albeit not significant, mean daily grades with amphetamine. No subject in either trial had a significantly lower mean daily headache grade on caffeine. CONCLUSIONS: Dextroamphetamine had real preventive effects on chronic tension-type and migraine headaches in some subjects. These results should encourage other investigators to study its effects on these headaches. PMID- 15546267 TI - Preventing disturbing migraine aura with lamotrigine: an open study. AB - BACKGROUND: Lamotrigine has been suggested as possibly effective for preventing migraine aura. OBJECTIVE: To describe our experience with a series of patients with disturbing migraine aura treated with lamotrigine. METHODS: The members of the Headache Group of the Spanish Society of Neurology were sent an ad hoc questionnaire to collect patients treated with lamotrigine due to disturbing migraine aura. The main outcome parameter ("response") was a >50% reduction in the mean frequency of migraine auras at 3 to 6 months of treatment. RESULTS: A total of 47 patients had been treated with lamotrigine due to severe migraine aura. Three could not complete the protocol as a result of developing skin rashes. Thirty (68%) patients responded. These were 21 females and 9 males whose ages ranged from 19 to 71 years. Eight suffered from migraine with "prolonged" aura, 8 typical aura with migraine headache (but had frequent episodes including speech symptoms), 6 basilar-type migraine, 6 typical aura without headache, and 2 hemiplegic migraine. Fifteen had been previously treated, without response, with other preventatives. The mean monthly frequency of migraine auras in these 30 patients changed from 4.2 (range: 1 to 15) to 0.7 (range: 0 to 6). Response was considered as excellent (>75% reduction) in 21 cases (70% of responders). Auras reappeared in 2 months in 9 out of 13 patients where lamotrigine was stopped, and ceased as soon as this drug was reintroduced. CONCLUSIONS: Lamotrigine should be considered in clinical practice for the preventive treatment of selected patients with disturbing migraine auras. Lamotrigine seems worthy of a controlled trial as prophylaxis of migraine aura. PMID- 15546269 TI - Multiple cranial neuropathies, headache, and facial pain in a septuagenarian. PMID- 15546270 TI - When can successful migraine prophylaxis be discontinued? PMID- 15546271 TI - Trigeminal neuralgia due to pontine infarction. AB - Trigeminal neuralgia (TN) is most commonly caused by vascular compression of the trigeminal nerve root entry zone. Secondary trigeminal neuralgia due to ischemic lesion of the pons is very rare. Here we report a patient with a pontine infarct transecting the central trigeminal pathways resulting with trigeminal neuralgia. PMID- 15546272 TI - Focal hypertrophic pachymeningitis in association with temporal arteritis. AB - We present an extremely rare case of a patient with focal hypertrophic pachymeningitis in association with histologically proven temporal arteritis (giant cell arteritis). It is proposed that focal hypertrophic pachymeningitis in itself is the contributing factor to severe headache rather than temporal arteritis alone. PMID- 15546273 TI - New onset migraine with aura following head injury: a case report. AB - A patient is described who developed recurrent headaches following a traumatic injury in the vicinity of the right greater occipital nerve (GON) that fulfill IHS criteria for migraine with aura. Both aura and headache can be triggered by pressure over the GON and relieved by local anesthetic blockade. PMID- 15546274 TI - Pharmacoeconomics: the cost of prophylactic migraine treatments. AB - Migraine preventive medications considered effective reduce headache frequency by 50 percent in approximately 50 percent of treated patients. In spite of similar effectiveness, these medications vary tremendonsly in their prices. Knowledge of medication prices and employing cost-effective strategies may greatly reduce treatment costs. PMID- 15546276 TI - Water deprivation headache: "new" variants of phenomenology. PMID- 15546277 TI - Ascertainment of photophobia and phonophobia during migraine: preferred close ended questions. PMID- 15546286 TI - Stress echocardiography: basics and noninvasive assessment of myocardial viability. AB - Myocardial viability within the infarct zone is an important determinant for left ventricular (LV) function recovery after interventional coronary revascularization. Echocardiographic techniques are highly valuable in identifying hibernation, especially in conjunction with dobutamine titration. Low doses may detect the inotropic reserve by significant enhancement of segmental wall motion abnormalities while high doses may surpass the ischemic threshold and wall motion deteriorates (biphasic response). According to the Task Forces on Clinical Application of Echocardiography by the American Heart Association (AHA) in cooperation with the American College of Cardiology (ACC), dobutamine echocardiography (DE) is officially recommended for the purpose of clinical decision making in respect of revascularization therapies (whether during the subacute infarct period or especially in chronic LV dysfunction due to coronary artery disease (CAD)). New methods such as strain rate imaging implemented in DE are on the way to give us quantitative measures of the amount of viability. PMID- 15546287 TI - Autologous skeletal myoblasts for myocardial regeneration. AB - We overview the current knowledge about the use of skeletal myoblasts in regeneration of infarcted myocardium. Myoblasts are attractive candidates for cell source for cardiomyoplasty in chronic postmyocardial injury as indicated by experimental and initial clinical experience. We also review the recent developments in skeletal myoblasts transplantation techniques with special attention to percutaneous transvenous approach to deliver therapeutic agents into myocardium from the lumen of coronary veins under intravascular guidance. PMID- 15546288 TI - Relationship between size of myocardial infarctions assessed by delayed contrast enhanced MRI after primary PCI, biochemical markers, and time to intervention. AB - BACKGROUND: The quantitative assessment of myocardial infarctions using delayed contrast-enhanced magnetic resonance imaging (MRI) has recently been validated using postmortem histopathological animal studies. In a prospective study, we investigated the correlation between infarct size as assessed by delayed contrast enhanced MRI, elevation of creatine kinase (CK), and c-reactive protein (CRP) as well as the time from onset of symptoms to intervention. METHODS: Four to 10 days after immediate PCI in 45 acute ST-segment elevation myocardial infarction (STEMI) patients (<24 hour) with stenting of the infarct-related artery and treatment with abciximab, we performed gadolinium contrast-enhanced three dimensional inversion recovery gradient-echo MR sequences with complete coverage of the LV-myocardium in short-axis slices. The mass of infarcted tissue based on the volume of hyperenhanced myocardium was calculated and linear regression analysis was performed to assess the correlation between absolute size of infarctions (g) as well as relative size (LV%) with peak values of CK, CRP, and the time to PCI. RESULTS: There was a significant correlation between absolute size of infarctions (g) and peak CK values (r = 0.72; P < 0.001) as well as the relative size (LV%) and peak CK (r = 0.77; P < 0.001). No correlations were found between absolute size (r = 0.33) as well as relative size (r = 0.27) of infarctions and peak CRP. There was also no correlation between absolute (r = 0.29) as well as relative size of infarctions (r = 0.27) and the time from onset of symptoms to PCI. CONCLUSIONS: In patients with acute STEMI (<24 hour) undergoing immediate PCI with stenting and treatment with abciximab, peak CK values correlated well with infarct size as assessed by delayed contrast-enhanced MRI. There were no correlations between infarct size and peak CRP as well as the time to intervention. PMID- 15546289 TI - Which parameter should be chosen as primary endpoint for randomized drug-eluting stent studies? AB - In Europe, 1,108 percutaneous coronary interventions (PCIs) per one Mio inhabitants are currently annually performed, most of them with stent implantation. Drug-eluting stents have been the focus of attention of interventional coronary therapy since the RAVEL study was first presented in September 2001 at the European Society of Cardiology Meeting. Ever since, numerous studies have assessed the effects of various antiproliferative and anti inflammatory substances and a variety of different stents was used as platform, either covered with polymer carriers of different chemical and physical properties or without a polymer carrier. CE- or FDA-certified drug-eluting stents are increasingly replacing the use of bare metal stents to reduce in-stent restenosis. Today, physicians have a choice of several approved drug-eluting stents and, therefore, need some evidence-based guidance through the "jungle of information" to make the right decisions. Even when focusing on randomized trials, differences between the studies regarding primary endpoints and sample sizes exist, making it difficult to compare the various drug-eluting stent studies. Randomized studies use either nonclinical (i.e., angiographic diameter stenosis, in-stent MLD, or in-stent late lumen loss) or clinical (i.e., TVF, TVR, and MACE) parameters as primary endpoints. Choosing an angiographic parameter as primary endpoint results in two major limitations: first, a significant improvement of an angiographic "surrogate" parameter does not necessarily translate into a better clinical outcome (DELIVER-I); second, conclusions regarding possible improvements of clinical outcome are underpowered, because the sample size calculation is based on the primary endpoint. Usually the number of patients needed is lower for angiographic than for clinical endpoints. Until today, only three trials with a primary clinical endpoint have shown a significantly positive impact on patients' outcome: the SIRIUS trial (Cypher stent) with its reduction of primary endpoint TVF (21.0% vs 8.6%), the TAXUS-IV trial (Taxus stent) with its reduction of primary endpoint TVR (12.0% vs 4.7%) and TAXUS-VI in long lesions with its reduction of primary endpoint TVR (19.4% vs 9.1%). Although the angiographic results of other drug-eluting stents are encouraging, they will have to prove their clinical impact based on adequately powered randomized trials with a primary clinical endpoint at an adequate time interval. PMID- 15546290 TI - Clinical experiences using everolimus-eluting stents in patients with coronary artery disease. AB - Despite the growing use of drug-eluting stents, restenosis remains to occur especially in high risk subgroups like patients with diffuse in-stent restenosis. This observation is supporting the search for new and potentially even more effective drug eluting stent systems. Everolimus has been used in conjunction with a new bioabsorbable polymer and gave promising results in initial clinical studies. In FUTURE I, a single-center, single-blinded randomized safety and feasibility study enrolling 15 patients with bare metal stents and 27 patients with everolimus-coated stents, 30-day MACE rate was 0% in both groups. In-stent late loss at six months was 0.83 mm in the control group and 0.10 mm in the everolimus group (p < 0.0001). In FUTURE II, a randomized multi-center study, a total of 64 patients were enrolled confirming safety and feasibility. After 6 months late loss was 0.85 mm in the control group and 0.12 mm in the everolimus group (p < 0.001). PMID- 15546291 TI - Drug-eluting bioabsorbable magnesium stent. AB - Current stent technology is based on the use of permanent implants that remain life long in the vessel wall, far beyond the time required for the prosthesis to accomplish its main goals of sealing dissection and preventing wall recoil. With the possibility to implant long vessel segments using antiproliferative drugs to prevent restenosis, the practice of transforming the coronary vessels into stiff tubes with a full metal jacket covering all side branches and being unable to adjust to the long-term wall changes, including wall remodeling with lumen ectasia becomes a serious concern. In this article, we describe the first biodegradable stent based on a magnesium alloy that allows controlled corrosion with release to the vessel wall and the blood stream of a natural body component such as magnesium with beneficial antithrombotic, antiarrhythmic, and antiproliferative properties. We also discuss the animal experiments and the initial clinical applications in 20 patients with implants below the knee, with final results soon to be released, and the plans for the first coronary study. The results of these last two studies will indicate whether the absence of a permanent implant and the antiproliferative properties shown in animals are sufficient to prevent the restenotic process in humans or whether the prosthesis must be modified by adding the biodegradable coating with conventional antiproliferative drugs. PMID- 15546292 TI - New approaches to ostial and bifurcation lesions. AB - Percutaneous coronary intervention of bifurcation lesions is associated with lower procedural success rates, and an increased subsequent rate of major adverse cardiac events and restenosis. Currently, an array of stenting possibilities suggests a rational approach to treat various bifurcation lesions with appropriate techniques. This is however seldom the case. The main problems of treating bifurcation lesions remain plaque shift leading to (threatened) side branch occlusion, and either too much or insufficient side branch ostial stent coverage predisposing to impaired side branch access or restenosis, respectively. This paper reviews the available technologies and their relative merits. PMID- 15546293 TI - New approaches to long lesions. AB - Long coronary lesions, comprising up to 20% of the interventional practice, pose a significant challenge to the interventionalist. Several approaches have been described to treat long coronary lesions with long-term suboptimal outcomes. Atherectomy and laser treatments with or without adjunctive balloon angioplasty have not been shown to be superior to conventional balloon angioplasty alone. Preliminary data with the use of drug-eluting stents in the treatment of long coronary lesions appears to be promising even in small vessels and may eventually become the mainstay treatment of long lesions. PMID- 15546294 TI - Novel approaches for the treatment of chronic total coronary occlusions. AB - Despite major advancements in the technology used for the percutaneous treatment of coronary artery disease, chronic total occlusions (CTOs) persist as a major challenge to the interventional cardiologist with relatively low success rates. CTOs are evident in 20% of patients undergoing cardiac catheterization and are responsible for the majority of cases that are referred to bypass surgery. There is growing evidence that patients may benefit from recanalization of a CTO by alleviation of angina, improving left ventricular function, and potentially long term survival. The major obstacle to percutaneous recanalization of CTOs is the inability to cross the occlusion with coronary guidewires. Even when crossed, the operator has to deal with the exact location of the distal wire (e.g., dissection or true lumen) and the existence of relatively long lesion requiring multiple stents with high restenosis rates. New technologies for CTO revascularization have been focused mainly on a mechanical approach including specialized guidewires and more recently, specific devices using highly sophisticated technology such as laser guidewire, optical coherence reflectometry, and a blunt microdissection catheter. An alternate biological approach involves the local administration of enzymes such as plasminogen activators (urokinase) or collagenase, which can act locally to specifically degrade the collagen content of the CTO, thereby "softening" the occlusion and allowing easier guidewire crossing. In conclusion, CTOs emerge as a great technical challenge and are the focus of novel series of mechanical and biological approaches. PMID- 15546295 TI - Carotid artery stenting: state of the art. AB - Carotid artery stenting (CAS) is growing as an alternative to carotid artery endarterectomy. Nowadays, it is performed routinely in many centers worldwide. Still, it is discussed controversially although several clinical trials have shown equivalency or superiority of catheter treatment at least in high-risk patients. What is still missing is a randomized trial in non-high-risk patients. This is an overview about the completed and ongoing trials as well as the current stent and embolic protection technology. PMID- 15546296 TI - Endovascular interventions in iliac and infrainguinal occlusive artery disease. AB - Percutaneous endovascular procedures are increasingly applied to treat symptomatic peripheral occlusive artery disease. While the primary technical success and recanalization rates in iliac and infrainguinal interventions are high, differences in the long-term patency rates exist with respect to the anatomic localization, separating the iliac, femoropopliteal, and infrapopliteal arterial regions. In iliac arteries, even complex lesions can be recanalized with good long-term patency rates, especially when using self-expanding nitinol stents. In the infrainguinal arteries the method of choice is still under debate (e.g., balloon angioplasty vs stent implantation). A high restenosis rate represents one of the major limitations in femoropopliteal and infrapopliteal interventions. Therefore, additional methods and treatment strategies for peripheral interventions with the potential for future applications are under investigation and will be discussed such as drug-eluting stents, brachytherapy, subintimal angioplasty, laser angioplasty, atherectomy/thrombectomy, cutting balloon, polytetrafluoroethylene (PTFE)-covered stent grafts, biodegradable stents, and cryoplasty. The increasing amount of data on successful peripheral interventions supports the necessity to adapt and reevaluate the current consensus guidelines that were put together in 2000. PMID- 15546297 TI - Abdominal aortic aneurysm: a review of endoluminal treatment. AB - Abdominal aortic aneurysm (AAA) carries a high mortality if left untreated. Until recently, most patients with AAA were treated with surgical repair. However, endoluminal graft treatment is rapidly becoming an alternative to surgical repair due to lower morbidity and comparable perioperative mortality rates. Despite this optimism patients and operators should keep in mind the palliative nature of these endovascular repairs and the lifelong need for surveillance, before embarking on these procedures. Endoleaks remain to be a significant problem leading to aneurysm expansion and occasionally rupture. Durability of the existing endograft devices remains to be seen. Careful patient selection is critical to success with these procedures. With future advancements in the endograft device technology, methods of patient surveillance, and patient care, there may be a shift from conventional surgical approaches to endovascular repair for the treatment of AAA. PMID- 15546298 TI - Influence of doxycycline on clinical and angiographic outcome following percutaneous coronary intervention. AB - BACKGROUND: Infection with Chlamydia pneumoniae may be involved in atherosclerosis. Neointima proliferation after percutaneous coronary intervention (PCI) could be aggravated by proinflammatory action of chronic vascular chlamydial infection. We investigated the influence of doxycycline therapy on restenosis rate and adverse clinical event rates after PCI. METHODS: We followed 1,019 consecutive patients in a single center nonrandomized observational study. During a 3-month period, all patients having PCI (n = 541; treatment group) received oral doxycycline 100 mg/day for 50 days starting from the day of the procedure. A control group (n = 478) consisted of consecutive post-PCI patients recruited from immediately preceding (n = 250) and ensuing periods. Event rates were gained by questionnaires after 259 +/- 57 days. Restenosis rate was derived from the routine angiographic restudies after 172 +/- 15 days. RESULTS: No significant differences between treatment and control group were found in adverse clinical event rates (myocardial infarction: 0.7% vs 1.3%; repeat-PCI: 24.0% vs 20.9%; coronary bypass grafting: 5% vs 3.9%; overall death after 6 months: 3.5% vs 2.1%) and overall restenosis rate (21.6% vs 23.4%). A subgroup analysis of nondiabetic male smokers revealed a lower restenosis rate in the treatment group (10.9% vs 30.9%; P = 0.0195). CONCLUSION: Doxycycline therapy for 50 days following PCI does not appear to have a substantial influence on the clinical and angiographic outcome in unselected patients. The observed reduction in restenosis rate among nondiabetic male smokers defines a new hypothesis that should stimulate further investigations. PMID- 15546299 TI - Transvenous pacing in infants: a faith based initiative? PMID- 15546300 TI - Endocardial pacemaker implantation in infants weighing < or = 10 kilograms. AB - Epicardial pacemaker implantation is the most common approach for small children requiring pacemaker implantation, though it is not free from complications. This article reviews the experience with endocardial pacemaker implantation, as an alternative approach, in children < or =10 kg at two centers. Thirty-nine children, median age 3.8 months (2 days-35 months), weight 4.6 kg (2.3-10 kg) underwent endocardial permanent pacing (VVI/R in 38, DDDR in 1). Indications for pacing were complete heart block (CHB) in 34 (congenital in 21, postsurgical in 12, congenitally corrected transposition of the great arteries 1), long QT syndrome in 3, and sinus bradycardia in 2 children. Two children with postsurgical CHB died 7 days and 3 weeks after implantation, respectively, due to heart failure and septicemia, despite appropriate pacemaker therapy. Over a median follow-up of 4.3 years (9 months-15.3 years), 12 patients underwent 18 generator replacements. Five patients were upgraded to physiological pacing. Ten patients underwent 12 ventricular lead advancements. Ventricular lead extraction was attempted 11 times in nine patients and succeeded 10 times. Two patients were converted to epicardial dual chamber systems. Two prepectorally placed generators required resiting due to threatened skin necrosis. Infective endocarditis on the lead, 9 months postimplant required removal of the system in one patient. The subclavian vein was found to be asymptomatically thrombosed in four patients. Endocardial permanent pacing is feasible and effective in children < or = 10 kg and an acceptable alternative to epicardial pacing. PMID- 15546301 TI - Stability of ventricular repolarization in conscious dogs with chronic atrioventricular dissociation and his-bundle pacing. AB - Following AVN ablation, eight dogs were surgically instrumented for chronic (continuous) HIS-bundle pacing. For data collection, implanted pacemakers were transiently programmed to pace in stepwise ascending ramps at rates from 50 to 200 beats/min in 30-beat/min steps. Each rate was held for 60 seconds. At each rate, ECG signals were collected from conscious dogs for measurements of QT intervals during the last 10 seconds of each paced rate to construct a QT-HR ramp. This QT-HR ramp was repeated twice on each day of study 10 minutes apart. Dogs were randomly assigned to two groups and studied weekly for a minimum of 6 weeks. Group 1 dogs had pacemakers programmed to a rate of 80 beats/min for the duration of the study. Group 2 dogs were paced at 80 beats/min for weeks 1-3, then increased to 140 beats/min for weeks 4-6. The difference between paired QT HR ramps within 1 day was <3 ms. QT-HR ramps were statistically indistinguishable over the 6-week study for group 1 dogs. Group 2 dogs experienced a slight flattening in the slope of the QT-HR ramps from week 3 to week 6 due to a reduction in QT interval at low HRs (50 and 80 beats/min) only. This conscious HIS-pacing model in dogs is a sensitive, stable, and reproducible method to define ventricular repolarization characteristics over a range of programmable HRs and experimental conditions. PMID- 15546302 TI - Autonomic modulation of the u wave during sympathomimetic stimulation and vagal inhibition in normal individuals. AB - Prolonged repolarization time, an important contributor to the pathogenesis of ventricular arrhythmias, is usually identified by a long QT interval (QT) on the ECG but is frequently confounded by the presence of a U wave. The physiological basis and clinical relevance of the U wave is unresolved. To better understand the relationship between the T and U waves, this study examined their behavior during nonresting autonomic conditions. Twenty-five healthy subjects were evaluated during sympathomimetic infusion with isoproterenol and vagal inhibition with atropine. As heart rate (HR) increased in response to isoproterenol, the QU interval (QU) decreased by an eightfold greater extent than QT. Furthermore, a marked increase in U wave amplitude and decrease in T wave amplitude were observed with T and U wave fusion at higher HRs. During atropine, QU decreased by only a threefold greater extent than QT, T and U wave amplitudes were affected only minimally, and T-U wave fusion was not observed. These results demonstrate that sympathomimetic stimulation causes striking alterations in the timing and amplitude of U waves that differ from effects on the T wave. These effects are not observed during vagal inhibition. Thus, the U wave represents a component of cardiac repolarization that is electrocardiographically and physiologically distinct from the T wave with a unique response to sympathomimetic stimulation. PMID- 15546303 TI - Direct comparison of a contractility and activity pacemaker sensor during treadmill exercise testing. AB - There are limited data about the chronotropic capacity of the peak endocardial acceleration (PEA) sensor. This study directly compared the chronotropic function from the PEA and the activity (ACT) sensor. The study included 18 patients (age 73 +/- 7 years) with > or = 75% pacemaker-driven heart rate (HR) and a PEA sensor and 11 healthy controls (age 67 +/- 7 years) underwent a chronotropic assessment exercise protocol (CAEP) exercise test with the pacemaker patients in VVIR mode after programming the sensors in the default setting with adjustment of the upper sensor rate as an age related maximum value (220-age). The ACT sensor was externally strapped on the thorax. Achieved exercise duration for the patients and controls was, respectively, 9.2 +/- 3 vs 18.4 +/- 4 minutes (P <0.001). The maximal achieved HR with the PEA sensor was 124 +/- 25 beats/min, versus the ACT with 140 +/- 23, versus the controls with 153 +/- 26 beats/min (P <0.001 between the groups). For the PEA, ACT, and controls, the time to peak HR was, respectively, 11 +/- 3, 7 +/- 3.6, and 18 +/- 4 (P <0.001 between groups) and HR after 10 minutes recovery was, respectively, 80 +/- 20, 65 +/- 15, and 82 +/- 4 beats/min (P <0.001 between groups). The PEA sensor functions hypochonotroop during exercise programmed as a single sensor system. It is, therefore, preferable to combine the PEA sensor with an activity-based sensor in a dual sensor system. Although both groups had normal left ventricular functions, the exercise capacity of pacemaker patients is significantly lower than in the controls. PMID- 15546304 TI - An epicardial subxiphoid implantable defibrillator lead: superior effectiveness after failure of standard implants. AB - A single epicardial implantable lead using the subxiphoid approach is described in this article. It consists of a single halo-shaped coil that is implanted under the inferior surface of the heart, including the right and left inferior ventricular surfaces. It has been implanted in four patients who could not be defibrillated with a transvenous system, even with the adjunct use of subcutaneous leads or left chest wall patch. Three of the patients had progressive heart failure due to ischemic myocardiopathy; the fourth patient had a dilated idiopathic myocardiopathy. The approach is simple and appears to be effective due to its ability to encompass the left and right ventricles. This vector seems to significantly lower the threshold for defibrillation, and may offer substantial benefit in the setting of high defibrillation thresholds with conventional leads, or when conventional systems are inadequate to achieve consistent defibrillation. PMID- 15546305 TI - Different clinical courses and predictors of atrial fibrillation occurrence after transisthmic ablation in patients with preablation lone atrial flutter, coexistent atrial fibrillation, and drug induced atrial flutter. AB - The aim of this prospective study was to compare the long-term follow-up after transisthmic ablation of patients with preablation lone atrial flutter, coexistent AF, and drug induced atrial flutter to determine if postablation AF followed a different clinical course and displayed different predictors in these groups. The study evaluated 357 patients who underwent transisthmic ablation for typical atrial flutter. These were divided into four groups according to their preablation history. Group A included patients with typical atrial flutter and without preablation AF (n=120, 33.6%). Group B included patients with preablation AF and spontaneous atrial flutter (n=132, 37.0%). Group C patients had preablation AF and atrial flutter induced by treatment with IC drugs (propafenone or flecainide) (n=63, 17.6%) Group D included patients with preablation AF and atrial flutter induced by treatment with amiodarone (n=42, 11.8%). During a mean follow-up of 15.2 double dagger 10.6 months (range 6-55 months) AF occurred more frequently in groups B (56.1%) and C (57.1%) patients than in groups A (20.8%, P <0.0001) and D (31.0%, P <0.0001) patients. The results of multivariate analysis revealed that different clinical and echocardiographical variables were correlated with postablation AF occurrence in the different groups. Patients with atrial flutter induced by amiodarone have a significantly lower risk of postablation AF than patients with spontaneous atrial flutter and AF, and those with atrial flutter induced by IC drugs. Different clinical and echocardiographical variables predict postablation AF occurrence in different subgroups of patients. PMID- 15546306 TI - Relationship of baseline electrocardiographic characteristics with the response to cardiac resynchronization therapy for heart failure. AB - Prospective identification of patients most and least likely to respond to cardiac resynchronization therapy (CRT) for congestive heart failure (CHF) will allow clinicians to target this intervention most efficiently. The authors hypothesized that ECG variables including RBBB and indicators of RV dysfunction and extensive prior myocardial infarction would correlate with diminished response to CRT. This study analyzed preimplantation ECGs in 110 patients with ICD indications and CHF due to left ventricular systolic dysfunction randomized to active biventricular pacing in the MIRACLE ICD trial. Clinical and ECG variables on the outcome of change in peak oxygen consumption from baseline to 6 months (Delta-VO2 ) were evaluated. For this cohort, average peak VO2 improved from 13.4 to 14.3 mL/kg per minute. Among clinical variables, the strongest predictors of increasing Delta-VO2 were reduced exercise time and peak VO2 at baseline. A dominant R wave in lead a VR, RBBB, and evidence of prior anterior infarction were each associated with significantly smaller average improvements in Delta-VO2 than their absence in univariate analysis. Alternative ECG criteria, including QRS duration, had no relationship with the outcome. In a multivariate model, only baseline VO2 (beta=-0.3, P=0.001) and ECG evidence of prior anterior infarction (beta=-1.3, P=0.03) were associated with the outcome. ECG markers of anterior infarction and RV dilation may help identify CHF patients unlikely to benefit from biventricular pacing. Further assessment is needed of these and other predictors of therapeutic response to CRT. PMID- 15546307 TI - Effect of amiodarone dose on the risk of permanent pacemaker insertion. AB - Bradyarrhythmia requiring permanent pacemaker insertion has been associated with amiodarone use but the effect of amiodarone dose has not been investigated. In order to determine the effect of amiodarone dose on the risk of requiring permanent pacemaker insertion, a cohort of 15,824 subjects with atrial fibrillation (AF) and prior myocardial infarction was established. This study included 1,340 subjects who received a first prescription of amiodarone at > 65 years of age. Cox regression with daily dose and cumulative dose (weighted for recency of exposure) represented by time dependent covariates was performed, adjusting for baseline risk factors and time dependent exposure to other cardiac medications. The incidence of pacemaker insertion was 2.2% per person-year during a mean of 1.8 +/- 1.5 years of follow-up, and 5.2% per person-year during the first 90 days of amiodarone exposure. Amiodarone daily doses > 200 mg were associated with an increased risk during the entire follow-up HR 2.0; 95% CI 1.0 to 4.1) as well as during the first 90 days (HR 3.1; 95% CI 1.1 to 9.0). Cumulative doses greater than the equivalent of continuous therapy with 200 mg per day were also associated with an increased risk (HR 2.8; 95% CI 1.4 to 5.5). Baseline conduction disorder or sinus node dysfunction was the only other significant predictor of pacemaker insertion. This study suggests that there is a dose dependent increased risk of permanent pacemaker insertion associated with amiodarone use that appears to be greatest during the initial months of treatment. PMID- 15546308 TI - Pacemakers and implantable cardioverter defibrillators: device longevity is more important than smaller size: the patient's viewpoint. AB - The size of pacemakers and implantable cardioverter defibrillators (ICDs) has been diminishing progressively. If two devices are otherwise identical in components, features and technology, the one with a larger battery should have a longer service life. Therefore, patients who receive smaller devices may require more frequent surgery to replace the devices. It is uncertain whether this tradeoff for smaller size is desired by patients. We surveyed 156 patients to determine whether patients prefer a larger, longer-lasting device, or a smaller device that is less noticeable but requires more frequent surgery. The effects of subgroups were evaluated; these included body habitus, age, gender, and patients seen at time of pulse generator replacement (PGR), initial implant, or follow-up. Among 156 patients surveyed, 151 expressed a preference. Of these, 90.1% preferred the larger device and 9.9% the smaller device (P <0.0001). Among thin patients, 79.5% preferred a larger device. Ninety percent of males and 89.2% of females selected the larger device. Among younger patients (< or =72 years), 89.6% preferred the larger device, as did 90.5% of older patients (>72 years). Of patients undergoing PGR or initial implants, 95% favored the larger device, as did 86% of patients presenting for follow-up. The vast majority of patients prefer a larger device to reduce the number of potential replacement operations. This preference crosses the spectrum of those with a previously implanted device, those undergoing initial implants, those returning for routine follow-up, and patients of various ages, gender, and habitus. PMID- 15546309 TI - Feasibility of catheter cryoablation in normal ventricular myocardium and healed myocardial infarction. AB - Although novel cryoablation systems have recently been introduced into clinical practice for catheter ablation of supraventricular tachycardia, the feasibility of catheter cryoablation of VT is unknown. Thus, the present study evaluates catheter cryoablation of the ventricular myocardium (1) in healthy sheep and (2) of VT in chronic myocardial infarction (MI). In three healthy sheep, 21 ventricular lesions (12 left and 9 right ventricle) were created with a catheter cryoablation system. Different freeze/thaw characteristics were used for lesion creation. The mean nadir temperature was -84.1 degrees C +/- 0.9 degrees C, mean lesion volume was 175.8 +/- 170.3 mm3, and 5 of 21 lesions were transmural. Lesion dimensions were 7.5 +/- 3.1 mm (width) and 4.2 +/- 2.5 mm (depth). Left ventricular lesions were significantly larger than right ventricular lesions (262 +/- 166 vs 60.5 +/- 91.6 mm3, P=0.0025). There was no difference in lesion volume with respect to different freeze/thaw characteristics. Anatomically (n=3) or electrophysiologically (n=3) guided catheter cryoablation was attempted in six sheep 105 +/- 56 days after MI, three of six animals had reproducibly inducible VT with a mean cycle length of 215 +/- 34 ms prior to ablation. In these animals, five VTs were targeted for ablation. A mean of 6 +/- 3 applications for nine left ventricular lesions were applied, six of nine lesions were transmural. The mean lesion volume was 501 +/- 424 mm3. No VT was inducible in two of three animals after cryoablation using an identical stimulation protocol. Therefore, catheter cryoablation of VT in healed MI is feasible, and no acute complications were observed. PMID- 15546310 TI - Evaluation of the early hemodynamic changes in carotid arteries during ventricular and dual chamber pacing. AB - In spite of a wide choice of pacemakers, there are some problems in making more rational clinical decisions for individual patients since mode selection and programming is usually performed on the basis of a clinical hunch. The aim of this study was to measure the differences in carotid flow in patients with a pacemaker programmed in the dual chamber and in the single chamber pacing modes. Sixty patients with implanted bipolar DDD pacemakers were enrolled in this study. Blood peak systolic velocity (PSV) and end-diastolic velocity (EDV), cross sectional area, resistive index (RI), and pulsatility index (PI) were measured in the common (CCA), internal (ICA), and external (ECA) carotid arteries before pacemaker implantation and after dual chamber and ventricular pacing at 60 beats/min. PSVs in the left CCA (79.3 +/- 24.9 cm/s) and right CCA (84.1 +/- 18.7) were shown to significantly decrease after VVI pacing (60.1 +/- 16.6 and 62.1 +/- 20.0, respectively). There was also a similar significant decrease in PSV in the left and right ICAs and ECAs. Besides PSV, RI, and PI in the left and right CCAs, ICAs, and ECAs significantly decreased after VVI pacing. There was no similar decrease after DDD pacing. Cross-sectional area and flow volume in the CCA, ICA, and ECA were similar after DDD and VVI pacing and before pacemaker implantation suggesting that cardiac output was similar when the measurements were recorded. Carotid artery PSVs, pulsatility, and RIs were found to be significantly decreased during VVI pacing compared to baseline and DDD pacing. The greater incidence of adverse cerebral outcomes in patients with VVI rather than DDD pacing may be partly due to decreased carotid PSVs. PMID- 15546311 TI - A case of paroxysmal supraventricular tachycardia. PMID- 15546312 TI - Pulmonary veins: anatomy, electrophysiology, tachycardia, and fibrillation. AB - Recent years have seen an enormous amount of experimental and clinical research into role of the pulmonary veins (PVs) in atrial fibrillation (AF). Advanced imaging techniques have confirmed the findings of earlier postmortem studies and added further dimension to our knowledge of PV anatomy. Such work is vital for an effective approach to successful ablation of AF. Detailed mapping studies suggest that reentry within the PVs is most likely responsible for their arrhythmogenicity, although focal or triggered activity cannot be excluded. Further work also implicates the posterior left atrium in the genesis of AF. Investigation into the interplay between the PVs and left atrium has led to a reevaluation of the mechanisms underlying AF and suggests that the PVs may play a role in both the initiation and maintenance of this arrhythmia. In order for electrophysiologists to further develop the technical approach to ablation of AF and improve the clinical outcomes, these crucial issues must be resolved. PMID- 15546313 TI - Postprandial variations in ST-segment in a patient with Brugada syndrome and partial gastrectomy. AB - A 74-year-old man with a history of partial gastrectomy presented with an electrocardiogram consistent with Brugada syndrome and marked meal related fluctuations in the ST segment. ST-segment elevation was prominently attenuated at 30 minutes and increased at 120 minutes after meals. Analysis of heart rate variability revealed a relationship between postprandial heightened parasympathetic activity and increase in Brugada-type ECG abnormality. A rapid postprandial increase in blood glucose may initially stimulate sympathetic nervous activity and secondarily increase parasympathetic tone. Food intake can be associated with fluctuations in ST-segment elevation in patients with the Brugada syndrome. PMID- 15546314 TI - T wave oversensing by a cardioverter defibrillator implanted in a patient with the Brugada syndrome. AB - Oversensing of electrical signals in recipients of ICDs is a common complication causing the delivery of inappropriate therapy. The Brugada syndrome has a characteristic ECG pattern with dynamic temporal changes in response to various physiological or environmental conditions. This case report presents a patient with the Brugada syndrome and intermittent T wave oversensing by an ICD due to changes in T wave amplitude which could not be resolved by device reprogramming and required repositioning of the electrode. PMID- 15546315 TI - Thebesian valve imaging with electron beam CT angiography: implications for resynchronization therapy. AB - We report visualization of a prominent coronary sinus os valve (Thebesian valve), by electron beam computed tomographic angiography, which impeded an endocardial approach to left ventricular pacing. Resynchronization therapy was therefore performed with an epicardial approach to left ventricular lead placement. Electron beam computed tomographic angiography can provide detailed information for coronary sinus instrumentation, including anomalies potentially affecting the approach to resynchronization therapy. PMID- 15546316 TI - Extraction of a defibrillator lead through an inferior vena cava filter. AB - We report an extraction of a chronic, previously cut and abandoned, active fixation implantable cardioverter-defibrillator lead through an inferior vena cava filter. A long workstation sheath that crossed the filter was used, allowing the lead and all hardware to be withdrawn through a single opening in the filter. PMID- 15546317 TI - Hypersensitive carotid sinus syndrome due to neurofibromatosis-1 and manifested by repeated episodes of syncope. AB - A 28-year-old patient with neurofibromatosis-1 presented with syncope. The exam demonstrated a mass adjacent and inferior to the right occiput that extended to the posterior lateral right-sided neck. Initial invasive and noninvasive testing was negative. Imaging of her head and neck demonstrated a large neurofibroma enveloping her right carotid sinus without vessel occlusion or evidence of malignancy. An event recorder documented asystole. A pacemaker was implanted to avoid the surgical morbidity of removing the neck mass. The patient has since been free of syncope. We believe neurofibromatosis-1 should be included in the differential of syncope. PMID- 15546318 TI - Subtotal innominate vein occlusion after unsuccessful pacemaker implantation for resynchronization therapy. AB - Cardiac resynchronization therapy is increasingly used to treat patients with poor left ventricular function and asynchronous left ventricular contraction. Generally, a transvenous approach is used for implantation of pacing leads. However, coronary sinus lead implantation may be associated with various problems that may result in abandoning of the procedure. In this report a patient is described with subtotal innominate vein occlusion after such a procedure who underwent video-assisted thoracoscopic surgery (VATS) for implantation of epicardial left ventricular pacemaker leads for resynchronization therapy. PMID- 15546319 TI - An unusual obstacle in lead extraction. AB - This case report deals with an unusual complication in the removal of an active fixation implantable cardioverter defibrillator (ICD) lead. We were not able to pass the stylet beyond the point of lead fracture and this was subsequently found to be due to the stylet passing between the electrode and the outer layer of the lead insulation. The lead was removed by rotation extraction of the entire lead. PMID- 15546323 TI - The prospective analysis of remote ICD monitoring. PMID- 15546325 TI - Early morning changes in QT/RR relationship in patients with inducible VT. PMID- 15546327 TI - Low back pain in adolescents: no evidence for activity restriction. PMID- 15546328 TI - Evidence for exercise therapy in the treatment of chronic disease based on at least three randomized controlled trials--summary of published systematic reviews. AB - Final evidence for the overall benefits of exercise therapy in the treatment/rehabilitation of specific chronic disease comes from randomized controlled trials (RCTs). This paper summarizes current evidence that is based on a systematic review including data from at least three RCTs with contrast for exercise only. The quality of specific RCTs as well as the quality of systematic reviews varies, the newest ones usually being of higher quality than the older ones. The most consistent finding of the studies is that aerobic capacity and muscular strength of patients can be improved without causing detrimental effects on disease progression. Severe complications during these carefully tailored programs were rare. The treatment periods and follow-up times of the majority of the RCTs are of a too short duration to document group differences in disease progression. However, exercise reduces disease-related symptoms in many diseases, such as osteoarthritis, asthma and chronic obstructive pulmonary disorder. Also, RCTs studying patients with coronary heart disease as well as patients with heart failure show that all-cause mortality is lower in exercisers than in controls. PMID- 15546329 TI - Returning athletes with severe low back pain and spondylolysis to original sporting activities with conservative treatment. AB - The purpose of this study was to clarify the efficacy of conservative treatment in athletes with severe low back pain and spondylolysis, especially focusing on returning to original sporting activities. One hundred and four athletes (96 males and eight females), who consulted our sports medicine clinic during the 11 year period between September 1991 and October 2002 because of low back pain with an apparent defect of the pars interarticularis on plain radiographs, were subjected to conservative treatment. The mean age of the patients was 20.7 years. Of all the patients, 40 (38.5%) discontinued sporting activities due to severe low back pain, and were conservatively treated with activity restriction and antilordotic lumbosacral bracing, aimed at relieving low back pain by achieving stability of the fracture partly associated with fibrous union. After their low back pain was markedly reduced, the brace was removed and then individual training to return to the original sporting activities was started. Thirty-five patients (87.5%) could return to their original sporting activities in an average of 5.4 months (range: 1.0-11.5 months) after the onset of treatment, and could continue the activities for at least 6 months despite non-bony union. These results suggest that the outcome of conservative treatment with activity restriction and bracing appears to be satisfactory in controlling symptoms and returning to original sporting activities. Randomized controlled trials or comparative follow-up studies are needed to confirm our results. PMID- 15546330 TI - Associations between activities and low back pain in adolescents. AB - The aim of this cross-sectional study was to explore associations between self reported leisure activities and low back pain (LBP). The material included all adolescents in eighth and ninth grade in two geographical areas in the school year 1996-1997. Eighty-eight adolescents participated (mean age 14.7 years), making the response rate 84%. LBP during the preceding year was reported by 57%. Sixty-six percent reported physical activity 3 times weekly or more. The median time spent on television or computer was 15 h. In multivariate analyses, LBP was inversely associated with time spent on physical activity, in particular with regular walking or bicycling. LBP was associated with the use of television or computer more than 15 h weekly, but not with the time spent reading. The results confirm studies showing inverse associations between walking and LBP. Future research should involve prospective studies of the potential effects of walking or bicycling and other specific activities on LBP. PMID- 15546331 TI - Relationship of leg muscle strength and knee function in the early period after anterior cruciate ligament reconstruction. AB - The purpose of this descriptive study was to assess the relationship between knee function and the strength testing performance of various leg muscles in the early period after anterior cruciate ligament reconstruction (ACLR). Twenty-four patients who had recent ACLR (seven females and 17 males, mean age=27 years) were tested for leg muscle strength and knee function 2 weeks after surgery. The concentric peak torques of the hip and knee extensors and knee flexors were measured at 210 degrees s(-1). In addition, the latter two muscle groups were measured at 60 degrees s(-1), and also isometrically (at a knee angle of 60 degrees). Knee function was assessed using a three-dimensional motion analysis system combined with a force platform. This system determined knee joint angles, torques and powers during level walking, stair ascent and stair descent. Pearson's correlation coefficients were calculated to assess the relationships between the injured knee function variables and the injured/uninjured strength ratios. The correlation coefficients ranged (in absolute value) from 0.03 to 0.81, and were generally higher for the stair tasks compared with level walking, and highest for the knee extensors (as compared with the knee flexors and hip extensors), and the strength test velocity appears much less important than the muscle group tested. These results contribute evidence indicating the importance of knee extensor strength to knee function in the early period after ACLR. PMID- 15546332 TI - Mild eccentric stretch injury in skeletal muscle causes transient effects on tensile load and cell proliferation. AB - The biomechanical and regenerative responses in rabbit tibialis anterior muscle following a single eccentric stretch injury were investigated 1, 3 and 7 days after traumatization. The stretch injury produced a transient biomechanical effect. On day 1, the load at failure was decreased in the injured muscles compared with the uninjured controls (P<0.05), but on days 3 and 7 significant biomechanical differences were no longer recorded between the injured and control muscles. In immunohistochemical analyses, no overt myonecrosis or connective tissue disruption was observed in any of the stretch-injured muscles. Yet satellite cells were activated to incorporate a thymidine analogue, bromodeoxyuridine indicating mitotic activity. They did not, however, produce muscle-specific proteins, i.e., they did not differentiate further. Furthermore, the intact appearing but mildly injured myofibers of the stretched muscles expressed both neonatal myosin and vimentin near the myotendinous junctions. Collectively, these results suggest that a single mild eccentric stretch of skeletal muscle which does not cause gross structural alterations results in a short-term decrease in tensile load and induces transient proliferation of satellite cells and fibroblasts together with expression of primitive proteins in myofibers. PMID- 15546333 TI - The role of exercise and gender for physical self-perceptions and importance ratings in Swedish university students. AB - The purpose of this study was to investigate how scores on the Physical Self Perception Profile (PSPP), including scores on the Perceived Importance Profile (PIP), were related to self-reported exercise frequency, duration, and gender in sample of Swedish university students. A total of 164 participants completed the PSPP, PIP, and a questionnaire focusing on frequency and duration of exercise. Exercise frequency, duration, and gender predicted best the PSPP sub-domains of Sport Competence and Physical Conditioning. Exercising more frequently, and for a longer time on each occasion was associated with higher PSPP and PIP scores. Women generally displayed lower PSPP scores than men. These results suggest that exercise professionals need to master a range of appropriate exercise strategies, since doubts concerning self-presentation may work against establishing a regular exercise routine. PMID- 15546334 TI - A 10-year follow-up study of pulmonary function in symptomatic elite cross country skiers--athletes and bronchial dysfunctions. AB - A high prevalence of asthma-like symptoms and abnormal bronchial reactivity to various stimuli has been reported among endurance athletes. This report presents the cases of three cross-country skiers who have shown a progressive development of airway obstruction during their sport careers. The observed decline in FEV1, FEV1/FVC and/or FVC (FEV, forced expiratory volume; FVC, forced expiratory vital capacity) was greater than expected from longitudinal regression equations describing the ageing effect on pulmonary function. The three subjects encountered abnormal respiratory discomfort during their sport activity which was not systematically associated with abnormal bronchial reactivity (either to methacholine or exercise hyperventilation). At the end of the follow-up periods (9-12 years), all three skiers presented objective signs of airflow limitations during intense exercise. This report suggests that repeated chronic hyperventilation of cold dry air in cross-country skiers for several years can induce permanent bronchial disorders which may include remodeling processes and induce ventilatory limitations during intense exercise. PMID- 15546336 TI - An update of the HLA genomic region, locus information and disease associations: 2004. AB - The human major histocompatibility (MHC) genomic region at chromosomal position 6p21 encodes the six classical transplantation HLA genes and many other genes that have important roles in the regulation of the immune system as well as in some fundamental cellular processes. This small segment of the human genome has been associated with more than 100 diseases, including common diseases--such as diabetes, rheumatoid arthritis, psoriasis, asthma and various autoimmune disorders. The MHC 3.6 Mb genomic sequence was first reported in 1999 with the annotation of 224 gene loci. The locus and allelic information of the MHC continue to be updated by identifying newly mapped expressed genes and pseudogenes based on comparative genomics, SNP analysis and cDNA projects. Since 1999, new innovations in bioinformatics and gene-specific functional databases and studies on the MHC genes have resulted in numerous changes to gene names and better ways to update and link the MHC gene symbols, names and sequences together with function, variation and disease associations. In this study, we present a brief overview of the MHC genomic structure and the recent information that we have gathered on the MHC gene loci via LocusLink at the National Centre for Biological Information (http://www.ncbi.nih.gov/.) and the MHC genes' association with various diseases taken from publications and records in public databases, such as the Online Mendelian Inheritance in Man and the Genetic Association Database. PMID- 15546337 TI - Identification of an epitope from the epithelial cell adhesion molecule eliciting HLA-A*2402-restricted cytotoxic T-lymphocyte responses. AB - Because the epithelial cell adhesion molecule (Ep-CAM) is expressed in almost all carcinomas and human leucocyte antigen (HLA)-A*2402 is the most common allele in many ethnic groups, including Japanese, the identification of peptide sequences, which elicit HLA-A*2402-restricted Ep-CAM-specific cytotoxic T-lymphocyte (CTL) responses, would facilitate specific immunotherapy for various histological types of carcinomas. An epitope was identified through the following steps: (i) computer-based epitope prediction from the amino acid sequence of Ep-CAM, (ii) major histocompatibility complex (MHC) stabilization assay to determine the affinity of the predicted peptide with HLA-A*2402 molecules, (iii) stimulation of CD8+ T cells with peptide-pulsed dendritic cells and (iv) testing the CTL specificity by means of enzyme-linked immunospot (ELISPOT) assays, CTL assays and MHC/peptide-tetramer staining. Peripheral CD8+ T cells of four of five healthy donors after three rounds of stimulation with the peptide Ep-CAM173-181 (RYQLDPKFI) secreted interferon-gamma in ELISPOT assays when exposed to the peptide. A CTL clone specific to the peptide efficiently lysed Ep-CAM-expressing cancer cell lines in an HLA-A*2402-restricted fashion. Endogenous processing and presentation of the peptide in a lung cancer cell line were confirmed by means of cold target inhibition assays. The CTL clone was also lytic to normal bronchial epithelial cells but to a lesser extent at low effector: target ratios. All these data suggest that the peptide-specific CTL responses may play some roles both in anti-cancer and autoimmune reactions. The peptide should prove useful to study anti-Ep-CAM CTL responses among population possessing HLA-A*2402. PMID- 15546338 TI - TNF-alpha and TGF-beta1 gene polymorphisms and renal allograft rejection in Koreans. AB - This study was performed in order to evaluate the association of tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) gene polymorphisms with renal allograft rejection in Koreans. Five TNF-alpha ( 1031T/C, -863C/A, -857C/T, -308G/A and -238G/A) and two TGF-beta1 (codon 10 T/C and codon 25 G/C) single-nucleotide polymorphism (SNP) sites were studied by using polymerase chain reaction (PCR) single-strand conformation polymorphism and PCR restriction fragment length polymorphism methods in 100 controls and 164 patients. The patients underwent renal transplantation, having one or more Human leukocyte antigen (HLA)-A, HLA-B and HLA-DR antigens mismatched with their donors. For the TGF-beta1 gene, we also studied the polymorphism of donors. The allele frequencies of each SNP site in controls were not different from those of patients. The frequency of TNF-alpha high-producer genotype, -308GA, and TGF beta1 lower (intermediate)-producer genotype, codon 10 CC and codon 25 GG, were significantly higher in patients with recurrent acute rejection episodes (REs), compared to those in patients with no or one RE. The highest risk group for developing recurrent REs showed the combination of TNF-alpha high- and TGF-beta1 lower-producer genotypes. Analysis of chronic renal allograft dysfunction (CRAD) revealed that TGF-beta1 high-producer genotype of donors, codon 10 TT/TC and codon 25 GG, is associated with CRAD especially in patients with recurrent REs. The highest risk group for developing CRAD showed the combination of recipient's TNF-alpha high- and donor's TGF-beta1 high-producer genotypes. These results would be useful for predicting high-risk group for acute rejection or CRAD in renal transplantation. PMID- 15546339 TI - Cytotoxic T-lymphocyte antigen-4-CT60 polymorphism in rheumatoid arthritis. AB - Cytotoxic T-lymphocyte antigen-4 (CTLA-4) is a functional candidate gene with susceptibility to rheumatoid arthritis (RA). The aim of this work was to investigate the possible influence of the recently described CT60A/G dimorphism of the CTLA-4 gene in the susceptibility to RA in Spanish patients. A total of 433 RA patients and 398 control subjects were included in the study. Genotyping of CTLA-4 CT60 was performed using two different methods: polymerase chain reaction restriction fragment length polymorphism system using an amplification created restriction site and a TaqMan 5'-allelic discrimination assay. In order to validate results obtained by different methods, a quality-control exercise was performed. No significant deviation in the distribution of the alleles or genotypes of the CT60 was found when we compared RA patient and control groups. In addition, no differences in CTLA-4 CT60 genotypic distribution was found when RA patients and controls were stratified by the presence or absence of the shared epitope. In conclusion, our results do not support an association between CT60A/G polymorphism and susceptibility to RA in the Spanish population, although the contribution of other positions located within the 3' region of the CTLA-4 gene to RA susceptibility cannot be discarded. PMID- 15546340 TI - Major histocompatibility complex haplotypes in Spanish immunoglobulin A deficiency patients: a comparative fine mapping microsatellite study. AB - The most consistent finding in Immunoglobulin A deficiency (IgAD) genetics is the presence of susceptibility factors located in the major histocompatibility complex (MHC). We have described the existence of at least two distinct susceptibility genes in the MHC present in different haplotypes. The aim of the present study was to locate with precision the susceptibility genes present in DR1- and DR7-positive haplotypes, taking advantage of their structural diversity, as opposed to the conserved nature of the DR3-extended susceptibility haplotype (DR3/B8), that hampers a more exhaustive scrutiny. A detailed analysis with 20 markers along the MHC in the 400 haplotypes present in 100 IgAD families, with special density at Class II locations, was performed to define the minimal shared susceptibility region present in all haplotypes carrying DR1 and, on the other hand, in all DR7-positive haplotypes. A comparison of the fine microsatellite allele structure of DR-extended haplotypes in the Spanish population with those described for Swedish and British families revealed no difference in DRB1*0101 and DRB1*0102 haplotypes between both populations. Our data suggest that the etiologic mutation present in DRB1*0101 and DRB1*0102 in North Europe (Sweden and UK) is missing in the Spanish DRB1*0101 haplotypes but is present in the DQB1/DRB1 region in DRB1*0102 haplotypes. The results obtained also indicated that the most likely susceptibility gene in the DR7 haplotypes is either DQA1 or DRB1. PMID- 15546341 TI - HLA class-I and class-II allele frequencies and two-locus haplotypes in Melanesians of Vanuatu and New Caledonia. AB - HLA class-I and class-II allele frequencies and two-locus haplotypes were examined in 367 unrelated Melanesians living on the islands of Vanuatu and New Caledonia. Diversity at all HLA class-I and class-II loci was relatively limited. In class-I loci, three HLA-A allelic groups (HLA-A*24, HLA-A*34 and HLA-A*11), seven HLA-B alleles or allelic groups (HLA-B*1506, HLA-B*5602, HLA-B*13, HLA B*5601, HLA-B*4001, HLA-B*4002 and HLA-B*2704) and four HLA-C alleles or allelic groups (HLA-Cw*04, HLA-Cw*01, HLA-Cw*0702 and HLA-Cw*15) constituted more than 90% of the alleles observed. In the class-II loci, four HLA-DRB1 alleles (HLA DRB1*15, HLA-DRB1*11, HLA-DRB1*04 and HLA-DRB1*16), three HLA-DRB3-5 alleles (HLA DRB3*02, HLA-DRB4*01 and HLA-DRB5*01/02) and five HLA-DQB1 alleles (HLA DQB1*0301, HLA-DQB1*04, HLA-DQB1*05, HLA-DQB1*0601 and HLA-DQB1*0602) constituted over 93, 97 and 98% of the alleles observed, respectively. Homozygosity showed significant departures from expected levels for neutrality based on allele frequency (i.e. excess diversity) at the HLA-B, HLA-Cw, HLA-DQB1 and HLA-DRB3/5 loci on some islands. The locus with the strongest departure from neutrality was HLA-DQB1, homozygosity being significantly lower than expected on all islands except New Caledonia. No consistent pattern was demonstrated for any HLA locus in relation to malaria endemicity. PMID- 15546342 TI - Immunogenetics of HLA null alleles: implications for blood stem cell transplantation. AB - The transplantation of haematopoietic stem cells is a potentially curative therapy for a variety of haematological and non-haematological diseases. Matching of donor and recipient for human leucocyte antigens (HLA) is pivotal for the success of blood stem cell transplantation. HLA null alleles are characterized by the lack of a serologically detectable product. Because serological HLA diagnostics are increasingly replaced by DNA-based typing methods considering only small regions of the genes, null alleles may be misdiagnosed as normally expressed variants. The failure to identify an HLA null allele as a non-expressed variant in the stem cell transplantation setting may result in an HLA mismatch that is highly likely to stimulate allogeneic T cells and to trigger graft-vs host disease. For some HLA null alleles, the translation into a truncated polypeptide chain seems possible, which thus might act as minor histocompatibility antigens. Because the prevalence of HLA null alleles may be around 0.3% or even higher, a screening strategy for HLA null alleles should, therefore, be implemented in the clinical laboratory. It may consist of the combination of serology and standard molecular typing techniques. As the standard molecular techniques are sometimes troublesome especially for characterizing the cytosine island at the 5' end of HLA class I exon 4 and need continuously be updated, an alternative approach may consist of sequencing all samples from genomic DNA for exons 2-3 or 4 (class I) or exon 2 (class II), including the adjacent intron splicing sites. This approach will detect 36/40 so far known non expressed variants and has the potential to easily uncover novel variants, thus essentially minimizing the risk of overlooking these challenging variants. PMID- 15546343 TI - Loss of heterozygosity in the HLA class I region in human pancreatic cancer. AB - An altered human leukocyte antigen (HLA) class I expression constitutes an important tumor-escape mechanism counteracting T-cell mediated immune responses. We utilized the technique of microsatellite analysis to characterize the loss of heterozygosity (LOH) in the HLA class I region in 24 samples of patients with human ductal pancreatic carcinoma. The expression of HLA class I and the infiltration by T cells were studied in parallel by standard immunohistochemistry. The present study demonstrates LOH in the HLA class I region in five patients with pancreatic carcinoma. Immunohistochemical analysis showed that pancreatic carcinomas were frequently characterized by a total or partial loss of expression in HLA class I antigen. The positive or negative LOH status corresponded with the expression analysis in eight cases. Reduction of HLA class I expression without LOH was found in 14 cases. Lymphocyte infiltration with CD3+, CD4+, and CD8+ T cells did not show significant differences between LOH-positive and LOH-negative tumors. In conclusion, LOH does not seem to be the only factor for the reduced expression of HLA class I antigen as well as for the T-cell infiltration in this type of tumor. PMID- 15546344 TI - HLA-Cw*1214 allele arisen via recombination between HLA-Cw*070201 and HLA Cw*120201. AB - Allelic polymorphism of the major histocompatibility complex arises mostly from gene conversion. Intralocus gene conversion usually involves limited fragments of DNA, whereas recombination involving large fragments of DNA is considered to be a rare event. During routine sequencing-based typing of donors for the National Marrow Donor Program, a new HLA-C allele was identified in a Caucasian donor. The allele, HLA-Cw*1214, proved to be the product of recombination between HLA Cw*070201 and HLA-Cw*120201. Exons 1, 2, the 3' end of exon 3 and exon 4 (with one mismatch) belong to HLA-Cw*120201, whereas part of exon 3 belongs to HLA Cw*070201. Sequencing with primers based in exon 2 and exon 3 showed that intron 2 of the new allele also belonged completely to HLA-Cw*1202. The recombination event apparently occurred within exon 3 with the first point of recombination somewhere between codons 92 and 134 and the second one between codons 157 and 181. PMID- 15546345 TI - Role of 14-bp deletion in the HLA-G gene in the maintenance of pregnancy. AB - Differential expression of human leukocyte antigens (HLAs) on trophoblast has been the focus of many studies, specially on extravillous cytotrophoblast cells, which migrates into the maternal uterine tissues. These invading cells do not express classical major histocompatibility complex class I (-A and -B) and class II molecules, along with low expression of HLA-C. HLA-G is the predominantly expressed antigen along with HLA-E. Hence, it is believed that expressed antigens may be involved in materno-fetal tolerance. In the present study, we have studied 14-bp deletion polymorphism in the exon-8 of the non-classical HLA-G antigen. There was no difference in the frequency of deletion/insertion polymorphism in fertile normal women and recurrent spontaneous abortion (RSA) women. However, the number of heterozygotes (-14b/+14b) were increased in RSA women. The probable mechanism for the increase of heterozygotes in recurrent fetal loss is discussed in light of soluble HLA-G. PMID- 15546346 TI - Nomenclature for factors of the HLA system, update August 2004. PMID- 15546347 TI - Nomenclature for factors of the HLA system, update September 2004. PMID- 15546348 TI - Host and non-host pathogens elicit different jasmonate/ethylene responses in Arabidopsis. AB - Arabidopsis does not support the growth and asexual reproduction of the barley pathogen, Blumeria graminis f. sp. hordei Bgh). A majority of germlings fail to penetrate the epidermal cell wall and papillae. To gain additional insight into this interaction, we determined whether the salicylic acid (SA) or jasmonate (JA)/ethylene (ET) defence pathways played a role in blocking barley powdery mildew infections. Only the eds1 mutant and NahG transgenics supported a modest increase in penetration success by the barley powdery mildew. We also compared the global gene expression patterns of Arabidopsis inoculated with the non-host barley powdery mildew to those inoculated with a virulent, host powdery mildew, Erysiphe cichoracearum. Genes repressed by inoculations with non-host and host powdery mildews relative to non-inoculated control plants accounted for two thirds of the differentially expressed genes. A majority of these genes encoded components of photosynthesis and general metabolism. Consistent with this observation, Arabidopsis growth was inhibited following inoculation with Bgh, suggesting a shift in resource allocation from growth to defence. A number of defence-associated genes were induced during both interactions. These genes likely are components of basal defence responses, which do not effectively block host powdery mildew infections. In addition, genes encoding defensins, anti microbial peptides whose expression is under the control of the JA/ET signalling pathway, were induced exclusively by non-host pathogens. Ectopic activation of JA/ET signalling protected Arabidopsis against two biotrophic host pathogens. Taken together, these data suggest that biotrophic host pathogens must either suppress or fail to elicit the JA/ET signal transduction pathway. PMID- 15546349 TI - Signaling requirements and role of salicylic acid in HRT- and rrt-mediated resistance to turnip crinkle virus in Arabidopsis. AB - Inoculation of turnip crinkle virus (TCV) on the resistant Arabidopsis ecotype Di 17 elicits a hypersensitive response (HR), which is accompanied by increased expression of pathogenesis-related (PR) genes. Previous genetic analyses revealed that the HR to TCV is conferred by HRT, which encodes a coiled-coil (CC), nucleotide-binding site (NBS) and leucine-rich repeat (LRR) class resistance (R) protein. In contrast to the HR, resistance to TCV requires both HRT and a recessive allele at a second locus designated rrt. Here, we demonstrate that unlike most CC-NBS-LRR R genes, HRT/rrt-mediated resistance is dependent on EDS1 and independent of NDR1. Resistance is also independent of RAR1 and SGT1. HRT/rrt mediated resistance is compromised in plants with reduced salicylic acid (SA) content as a consequence of mutations eds5, pad4, or sid2. By contrast, HR is not affected by mutations in eds1, eds5, pad4, sid2, ndr1, rar1, or sgt1b. Resistance to TCV is restored in both SA-deficient Di-17 plants expressing the nahG transgene and mutants containing the eds1, eds5, or sid2 mutations by exogenous application of SA or the SA analog benzo(1,2,3)thiadiazole-7-carbothioic acid (BTH). In contrast, SA/BTH treatment failed to enhance resistance in HRT pad4, Col-0, or hrt homozygous progeny of a cross between Di-17 and Col-0. Thus, HRT and PAD4 are required for SA-induced resistance. Exogenously supplied SA or high endogenous levels of SA, due to the ssi2 mutation, overcame the suppressive effects of RRT and enhanced resistance to TCV, provided the HRT allele was present. High levels of SA upregulate HRT expression via a PAD4-dependent pathway. As Col-0 transgenic lines expressing high levels of HRT were resistant to TCV, but lines expressing moderate to low levels of HRT were not, we conclude that SA enhances resistance in the RRT background by upregulating HRT expression. These data suggest that the HRT-TCV interaction is unable to generate sufficient amounts of SA required for a stable resistance phenotype, and the presence of rrt possibly corrects this deficiency. PMID- 15546350 TI - The chromatin remodelling complex FACT associates with actively transcribed regions of the Arabidopsis genome. AB - The packaging of the genomic DNA into chromatin in the cell nucleus requires machineries that facilitate DNA-dependent processes such as transcription in the presence of repressive chromatin structures. Using co-immunoprecipitation we have identified in Arabidopsis thaliana cells the FAcilitates Chromatin Transcription (FACT) complex, consisting of the 120-kDa Spt16 and the 71-kDa SSRP1 proteins. Indirect immunofluorescence analyses revealed that both FACT subunits co-localize to nuclei of the majority of cell types in embryos, shoots and roots, whereas FACT is not present in terminally differentiated cells such as mature trichoblasts or cells of the root cap. In the nucleus, Spt16 and SSRP1 are found in the cytologically defined euchromatin of interphase cells independent of the status of DNA replication, but the proteins are not associated with heterochromatic chromocentres and condensed mitotic chromosomes. FACT can be detected by chromatin immunoprecipitation over the entire transcribed region (5' UTR, coding sequence, 3'-UTR) of actively transcribed genes, whereas it does not occur at transcriptionally inactive heterochromatic regions and intergenic regions. FACT localizes to inducible genes only after induction of transcription, and the association of the complex with the genes correlates with the level of transcription. Collectively, these results indicate that FACT assists transcription elongation through plant chromatin. PMID- 15546351 TI - The nuclear genes Lhcb and HEMA1 are differentially sensitive to plastid signals and suggest distinct roles for the GUN1 and GUN5 plastid-signalling pathways during de-etiolation. AB - Feedback mechanisms are critical to the regulation of chloroplast development and signals from functional plastids are required to maintain nuclear gene expression of chloroplast proteins. To understand the role of these signals in de-etiolating Arabidopsis thaliana L. seedlings, we followed the expression of three nuclear genes, Lhcb, HEMA1 and GSA, under a variety of treatments (Norflurazon, lincomycin and a far-red light pre-treatment) leading to plastid damage in white light and in a range of genetic backgrounds known to modulate plastid signalling: the genomes uncoupled mutants, gun1, gun4, gun5 and the gun1,5 double mutant, and in a transgenic line over-expressing NADPH:protochlorophyllide oxidoreductase. The three nuclear genes were differentially sensitive to changes in plastid signalling, with Lhcb the most strongly repressed and GSA insensitive to all but the most severe treatments. Analysis of plastid morphology in seedlings grown under identical conditions demonstrated that these responses corresponded closely to the degree of plastid damage. Furthermore, although Lhcb and HEMA1 were responsive to both GUN1 and GUN5 signals, the relative inputs from these pathways differed for each transcript with GUN1 being dominant for HEMA1 regulation. Further analysis of HEMA1 expression in gun1 seedlings under non-photobleaching conditions indicates that GUN1 is an important suppressor of HEMA1 expression in the dark and under saturating white light. These results are consistent with plastid signals functioning in a feedback regulatory mechanism during chloroplast biogenesis, and suggest a key role for GUN1 during the early stages of chloroplast development. PMID- 15546352 TI - The Arabidopsis COW1 gene encodes a phosphatidylinositol transfer protein essential for root hair tip growth. AB - Root hairs are a major site for the uptake of water and nutrients into plants, and they form an increasingly important model system for the study of development in higher plants. We now report on the molecular genetic analysis of the srh1 mutant in Arabidopsis thaliana impaired in root hair tip growth. We show that srh1 is a new allele of cow1 (can of worms1) and we identified the COW1 gene using a positional cloning strategy. The N-terminus of the COW1 protein is 32% identical to an essential phosphatidylinositol transfer protein (PITP), the yeast Sec14 protein (sec14p) while the C-terminus is 34.5% identical to a late nodulin of Lotus japonicus, Nlj16. We show that expression of the COW1 lipid-binding domain complements the growth defect associated with Sec14p dysfunction in yeast. In addition, we show that GFP fused to the COW1 protein specifically accumulates at the site of root hair outgrowth. We conclude that the COW1 protein is a PITP, essential for proper root hair growth. PMID- 15546353 TI - Molecular characterization of the tobacco SET domain protein NtSET1 unravels its role in histone methylation, chromatin binding, and segregation. AB - Plants contain a great number of genes encoding a distinctive class of SET domain proteins which harbor a plant-specific N-terminal part together with a C-terminal part showing highest sequence similarity to the catalytic domain of the yeast CLR4, the human SUV39H1 and G9a histone-methyltransferases (HMTases). Here we show that NtSET1, a representative member of this class from tobacco, methylated both K9 and K27 of histone H3 in vitro. Ectopic expression of NtSET1, by an inducible promoter, increased the amount of dimethylated H3K9 and induced chromosome-segregation defects in tobacco BY2 cells. Deletion analyses show that the HMTase activity, the association with specific chromatin regions and with condensed chromosomes, and the cellular effects largely depended on the C terminal region including the SET domain of the protein. Nevertheless, the N terminal part of NtSET1 was capable of targeting the green fluorescent protein to interphase chromatin. Finally, we show that NtSET1 bound LHP1, the Arabidopsis homolog of animal heterochromatin protein 1 (HP1), and that LHP1 co-localized with heterochromatin containing high amounts of dimethylated H3K9, suggesting a role for NtSET1 in heterochromatic function. Taken together, our results provide new insights into the molecular and global chromatin-binding activities of this particular class member of plant SET domain proteins. PMID- 15546354 TI - Mutation of a UDP-glucose-4-epimerase alters nematode susceptibility and ethylene responses in Arabidopsis roots. AB - In Arabidopsis, mutation of RHD1, a UDP-glucose-4-epimerase, causes root-specific phenotypes, including hypersusceptibility to the cyst nematode Heterodera schachtii, increased root hair elongation, decreased root length, and root epidermal bulging. Previous experiments suggested that increased ethylene sensitivity or production mediated the rhd1-4 phenotypes. In the present study, double mutant analyses revealed that only rhd1-4 hypersusceptibility to H. schachtii and increased root hair elongation were dependent upon the ethylene signaling genes EIN2 and EIN3 but not upon ethylene signaling mediated by the auxin efflux carrier EIR1. In contrast, the rhd1-4 short root and root epidermal bulging phenotypes did not require EIN2, EIN3, or EIR1. A time-course analysis of RHD1 transcript levels in wild-type plants treated with the ethylene precursor 1 aminocyclopropane-1-carboxylic acid showed a root-specific downregulation of RHD1 expression by ethylene. This observation was corroborated by our finding of increased RHD1 transcript levels in roots of the ethylene-insensitive mutants etr1 and ein2. In addition to ethylene, auxin strongly influences H. schachtii susceptibility and root hair elongation. Therefore, we investigated the sensitivity of rhd1-4 roots to indole-3-acetic acid (IAA). Equivalent IAA concentrations caused a greater reduction in rhd1-4 root elongation compared with wild-type roots. Finally, H. schachtii parasitism was found to strongly downregulate RHD1 expression in the root 3 days after inoculation. We conclude that RHD1 is a likely target of root-specific negative regulation by ethylene and that loss of RHD1 function results in a heightened sensitivity of root tissues to both ethylene and auxin. PMID- 15546355 TI - Conservation of the microstructure of genome segments in Brassica napus and its diploid relatives. AB - The cultivated Brassica species are the group of crops most closely related to Arabidopsis thaliana (Arabidopsis). They represent models for the application in crops of genomic information gained in Arabidopsis and provide an opportunity for the investigation of polyploid genome formation and evolution. The scientific literature contains contradictory evidence for the dynamics of the evolution of polyploid genomes. We aimed at overcoming the inherent complexity of Brassica genomes and clarify the effects of polyploidy on the evolution of genome microstructure in specific segments of the genome. To do this, we have constructed bacterial artificial chromosome (BAC) libraries from genomic DNA of B. rapa subspecies trilocularis (JBr) and B. napus var Tapidor (JBnB) to supplement an existing BAC library from B. oleracea. These allowed us to analyse both recent polyploidization (under 10,000 years in B. napus) and more ancient polyploidization events (ca. 20 Myr for B. rapa and B. oleracea relative to Arabidopsis), with an analysis of the events occurring on an intermediate time scale (over the ca. 4 Myr since the divergence of the B. rapa and B. oleracea lineages). Using the Arabidopsis genome sequence and clones from the JBr library, we have analysed aspects of gene conservation and microsynteny between six regions of the genome of B. rapa with the homoeologous regions of the genomes of B. oleracea and Arabidopsis. Extensive divergence of gene content was observed between the B. rapa paralogous segments and their homoeologous segments within the genome of Arabidopsis. A pattern of interspersed gene loss was identified that is similar, but not identical, to that observed in B. oleracea. The conserved genes show highly conserved collinearity with their orthologues across genomes, but a small number of species-specific rearrangements were identified. Thus the evolution of genome microstructure is an ongoing process. Brassica napus is a recently formed polyploid resulting from the hybridization of B. rapa (containing the Brassica A genome) and B. oleracea (containing the Brassica C genome). Using clones from the JBnB library, we have analysed the microstructure of the corresponding segments of the B. napus genome. The results show that there has been little or no change to the microstructure of the analysed segments of the Brassica A and C genomes as a consequence of the hybridization event forming natural B. napus. The observations indicate that, upon polyploid formation, these segments of the genome did not undergo a burst of evolution discernible at the scale of microstructure. PMID- 15546356 TI - Engineering of ubiquinone biosynthesis using the yeast coq2 gene confers oxidative stress tolerance in transgenic tobacco. AB - Ubiquinone (UQ), an electron carrier in the respiratory chain ranging from bacteria to humans, shows antioxidative activity in vitro, but its physiological role in vivo is not yet clarified in plants. UQ biosynthesis was modified by overexpressing the yeast gene coq2, which encodes p hydroxybenzoate:polyprenyltransferase, to increase the accumulation of UQ-6 in yeast and UQ-10 in tobacco. The yeast and tobacco transgenic lines showed about a three- and six-fold increase in UQ, respectively. COQ2 polypeptide, the localization of which was forcibly altered to the endoplasmic reticulum, had the same or a greater effect as mitochondria-localized COQ2 on the increase in UQ in both the yeast and tobacco transformants, indicating that the UQ intermediate is transported from the endoplasmic reticulum to the mitochondria. Plants with a high UQ level are more resistant to oxidative stresses caused by methyl viologen or high salinity. This is attributable to the greater radical scavenging ability of the transgenic lines when compared with the wild type. PMID- 15546357 TI - The effect of intron location on intron-mediated enhancement of gene expression in Arabidopsis. AB - Introns are often required for full expression of genes in organisms as diverse as plants, insects, nematodes, yeast, and mammals. To explore the potential mechanisms of intron-mediated enhancement in Arabidopsis thaliana, the effect of varying the position of an intron was determined using a series of reporter gene fusions between TRYPTOPHAN BIOSYNTHESIS1 (TRP1) and GUS. Two introns that differ in the degree to which they stimulate expression were individually tested at six locations within coding sequences and two positions in the 3'-UTR. The ability of the first introns from both the TRP1 and POLYUBIQUITIN10 (UBQ10) genes to elevate mRNA accumulation in transgenic plants was found to decline with distance from the promoter, despite their being efficiently spliced from all coding sequence locations. Neither intron significantly enhanced mRNA accumulation when positioned 1.1 kb or more from the start of transcription. In addition, measurements of GUS enzyme activity revealed that both introns at all locations elevated GUS activity more than they enhanced mRNA accumulation. The stimulation mediated by two of four other introns tested at the position nearest the promoter was also greater at the level of GUS activity than mRNA accumulation. These findings support a model in which introns increase transcription and promote translation by two distinct mechanisms. PMID- 15546358 TI - DNA array analyses of Arabidopsis thaliana lacking a vacuolar Na+/H+ antiporter: impact of AtNHX1 on gene expression. AB - AtNHX1, a vacuolar cation/proton antiporter of Arabidopsis, plays an important role in salt tolerance, ion homeostasis and development. We used the T-DNA insertional mutant of AtNHX1 (nhx1 plants) and Affymetrix ATH1 DNA arrays to assess differences in transcriptional profiles and further characterize the roles of a vacuolar cation/proton antiporter. Mature, soil-grown plants were used in this study to approximate typical physiological growing conditions. A comparison of plants grown in the absence of salt stress yielded many transcripts that were affected by the absence of the AtNHX1 vacuolar antiporter. Furthermore, changes in gene expression due to a non-lethal salt stress (100 mm NaCl) in the nhx1 plants were significantly different from the changes seen in wild-type plants. The nhx1 transcriptome was differentially affected when the plants were grown in the absence or presence of salt. In conclusion, in addition to the known role(s) of AtNHX1 on ion homeostasis, the vacuolar cation/proton antiporter plays a significant role in intracellular vesicular trafficking, protein targeting, and other cellular processes. PMID- 15546359 TI - The IAA1 protein is encoded by AXR5 and is a substrate of SCF(TIR1). AB - Recent studies of auxin response have focused on the functions of three sets of proteins: the auxin (Aux) response factors (ARFs), the Aux/IAAs, and the F-box protein TIR1. The ARF proteins bind DNA and directly activate or repress transcription of target genes while the Aux/IAA proteins repress ARF function. TIR1 is part of a ubiquitin protein ligase required for degradation of Aux/IAA proteins. Here we report the isolation and characterization of a novel mutant of Arabidopsis called axr5-1. Mutant plants are resistant to auxin and display a variety of auxin-related growth defects including defects in root and shoot tropisms. Further, the axr5-1 mutation results in a decrease in auxin-regulated transcription. The molecular cloning of AXR5 revealed that the gene encodes the IAA1 protein, a member of the Aux/IAA family of proteins. AXR5 is expressed throughout plant development consistent with the pleiotropic mutant phenotype. The axr5-1 mutation results in an amino acid substitution in conserved domain II of the protein, similar to gain-of-function mutations recovered in other members of this gene family. Biochemical studies show that IAA1/AXR5 interacts with TIR1 in an auxin-dependent manner. The mutation prevents this interaction suggesting that the mutant phenotype is caused by the accumulation of IAA1/AXR5. Our results provide further support for a model in which most members of the Aux/IAA family are targeted for degradation by SCFTIR1 in response to auxin. PMID- 15546360 TI - Functional differentiation of endosomes in Arabidopsis cells. AB - Endocytosis plays an important role in plant physiology, but how endocytic organelles are organized remains unknown. We present the evidence that endosomes are functionally differentiated in Arabidopsis cells. Two types of Rab5-related GTPases are localized on distinct population of endosomes in a partially overlapping manner. Ara7 and Rha1 are on an early type of endosomes with AtVamp727, where recycling of plasma membrane proteins occurs. In contrast, the plant-unique Rab5, Ara6, resides on distinct endosomes with the prevacuolar SNAREs. Partially overlapping localization of Ara6 and Ara7/Rha1 with reciprocal gradients suggests maturation of endosomes from one to the other. PMID- 15546361 TI - The Pseudomonas syringae type III effector AvrRpt2 promotes virulence independently of RIN4, a predicted virulence target in Arabidopsis thaliana. AB - AvrRpt2, an effector protein from Pseudomonas syringae pv. tomato (Pst), behaves as an avirulence factor that activates resistance in Arabidopsis thaliana lines expressing the resistance gene RPS2. AvrRpt2 can also enhance pathogen fitness by promoting the ability of the bacteria to grow and to cause disease on susceptible lines of A. thaliana that lack functional RPS2. The activation of RPS2 is coupled to the AvrRpt2-induced disappearance of the A. thaliana RIN4 protein. However, the significance of this RIN4 elimination to AvrRpt2 virulence function is unresolved. To clarify our understanding of the contribution of RIN4 disappearance to AvrRpt2 virulence function, we generated new avrRpt2 alleles by random mutagenesis. We show that the ability of six novel AvrRpt2 mutants to induce RIN4 disappearance correlated well with their avirulence activities but not with their virulence activities. Moreover, the virulence activity of wild type AvrRpt2 was detectable in an A. thaliana line lacking RIN4. Collectively, these results indicate that the virulence activity of AvrRpt2 in A. thaliana is likely to rely on the modification of host susceptibility factors other than, or in addition to, RIN4. PMID- 15546362 TI - Chloride fluxes in lily pollen tubes: a critical reevaluation. AB - Microelectrodes, made from a Cl(-)-selective liquid ion exchanger previously used to measure putative Cl- fluxes in Lilium longiflorum pollen tubes, were characterized. The electrodes were poorly selective, possessing only about 10 fold selectivity for Cl- over other anions tested. They had only 2.4-fold selectivity for Cl- over the anionic form of the H+ buffer, MES, indicating that the electrode can indirectly detect H+ gradients. Apparent anion influx was detected along the pollen tube shafts and at the grains while apparent anion efflux was detected near the tip of the tube. During oscillating growth, the peak of the oscillating apparent anion efflux at the tip occurred, on average, 7.9 sec after the peak of the growth oscillations. Consideration of the previously characterized H+ fluxes in lily pollen grains and tubes, as well as the poor anion selectivity of the Cl- electrodes, indicates that the putative Cl- fluxes are in fact changes in the anionic concentration of the buffer resulting from H+ gradients and not changes in Cl- concentration. The claim of a central role for Cl- in lily pollen tube growth is further undermined by the fact that these tubes grow at the same rate if the Cl- content of the growth medium is reduced to trace levels (< or =31 microM), and that the grains have only small reserves of Cl-. These results lead to the conclusion that Cl- fluxes are not a significant component of pollen tube growth and Cl- itself is not required for growth. PMID- 15546363 TI - ACC synthase expression regulates leaf performance and drought tolerance in maize. AB - Ethylene regulates entry into several types of plant developmental cell death and senescence programs besides mediating plant responses to biotic and abiotic stress. The response of cereals to conditions of drought includes loss of leaf function and premature onset of senescence in older leaves. In this study, ACC synthase (ACS) mutants, affecting the first step in ethylene biosynthesis, were isolated in maize and their effect on leaf function examined. Loss of ZmACS6 expression resulted in delayed leaf senescence under normal growth conditions and inhibited drought-induced senescence. Zmacs6 leaves continued to be photosynthetically active under both conditions indicating that leaf function was maintained. The delayed senescence phenotype associated with loss of ZmACS6 expression was complemented by exogenous ACC. Surprisingly, elevated levels of foliar chlorophyll, Rubisco, and soluble protein as well as improved leaf performance was observed for all Zmasc6 leaves, including young and fully expanded leaves which were far from initiating senescence. These observations suggest that ethylene may serve to regulate leaf performance throughout its lifespan as well as to determine the onset of natural senescence and mediate drought-induced senescence. PMID- 15546364 TI - Hypocotyl growth orientation in blue light is determined by phytochrome A inhibition of gravitropism and phototropin promotion of phototropism. AB - How developing seedlings integrate gravitropic and phototropic stimuli to determine their direction of growth is poorly understood. In this study we tested whether blue light influences hypocotyl gravitropism in Arabidopsis. Phototropin1 (phot1) triggers phototropism under low fluence rates of blue light but, at least in the dark, has no effect on gravitropism. By analyzing the growth orientation of phototropism-deficient seedlings in response to gravitropic and phototropic stimulations we show that blue light not only triggers phototropism but also represses hypocotyl gravitropism. At low fluence rates of blue light phot1 mutants were agravitropic. In contrast, phyAphot1 double mutants grew exclusively according to gravity demonstrating that phytochrome A (phyA) is necessary to inhibit gravitropism. Analyses of phot1cry1cry2 triple mutants indicate that cryptochromes play a minor role in this response. Thus the optimal growth orientation of hypocotyls is determined by the action of phyA-suppressing gravitropism and the phototropin-triggering phototropism. It has long been known that phytochromes promote phototropism but the mechanism involved is still unknown. Our data show that by inhibiting gravitropism phyA acts as a positive regulator of phototropism. PMID- 15546365 TI - In vitro culture of Arabidopsis embryos within their ovules. AB - Embryogenesis of flowering plants establishes a basic body plan with apical basal, radial and bilateral patterns from the single-celled zygote. Arabidopsis embryogenesis exhibits a nearly invariant cell division pattern and therefore is an ideal system for studies of early plant development. However, plant embryos are difficult to access for experimental manipulation, as they develop deeply inside maternal tissues. Here we present a method for the culture of zygotic Arabidopsis embryos in vitro. The technique omits excision of the embryo by culturing the entire ovule, thus greatly facilitating the time and effort involved. It enables external manipulation of embryo development and culture from the earliest developmental stages up to maturity. Administration of various chemical treatments as well as the use of different molecular markers is demonstrated together with standard techniques for visualizing gene expression and protein localization in in vitro cultivated embryos. The presented set of techniques allows for so far unavailable molecular physiology approaches in the study of early plant development. PMID- 15546367 TI - A novel method to estimate pulmonary artery wedge pressure using the downslope of the Doppler mitral regurgitant velocity profile. AB - Continuous-wave (CW) Doppler recording of mitral regurgitation (MR) is a reflection of the left ventriculoatrial pressure gradient. Accordingly, this jet may yield information about pulmonary artery wedge pressure (PAWP). In this study, we derived and then evaluated a novel method for prediction of PAWP. Patients (n=80) with moderate to severe MR and left ventricular dysfunction were included in the study. Transthoracic echocardiography was performed in patients during pulmonary artery pressure monitoring. A satisfactory CW Doppler recording of MR was obtained in 63/80 (78%). On the late descending portion of the CW recording, the time from a velocity of 4 m/sec to the end of the jet was defined as t1, and from 3 m/sec to the end of the jet as t2. Mathematical derivation of t1/t2 as a predictor of PAWP, was performed based on Weiss' derivation. If t1/t2 was <1.30, the PAWP was normal. If t1/t2 > 1.44, the PAWP was > 16 mmHg. With this new mathematical derivation, it appears that the downslope of the CW Doppler MR waveform may be able to distinguish a normal from elevated PAWP. PMID- 15546368 TI - Inducible left ventricular outflow tract gradient during dobutamine stress echocardiography: an association with intraoperative hypotension but not a contraindication to liver transplantation. AB - BACKGROUND: Dobutamine stress echocardiography (DSE) is frequently used in the evaluation of cardiac risk prior to orthotopic liver transplantation (OLT). In the general cardiac population, an inducible left ventricular outflow tract gradient (LVOT Delta) during DSE has variable prognostic importance. The purpose of this study was to determine the prevalence and clinical significance of LVOT Delta in patients undergoing OLT during DSE. METHODS: Consecutive medical records of 106 patients who had undergone OLT at our institution from January 1997 until January 2002 were retrospectively analyzed and divided into two groups based on the presence (Group I, LVOT Delta >36 mmHg) or absence (Group II, LVOT Delta< or = 36 mmHg) of a significant LVOT Delta measured during DSE. We determined any outcome differences between these two groups with regard to intraoperative hypotension, cardiac mortality, length of hospital stay, graft function, and renal function post-OLT. RESULTS: Forty-six patients had an LVOT Delta > 36 mmHg (Group I) and 60 patients had LVOT Delta< or = 36 mmHg (Group II). Baseline demographics were similar in both groups. There was no significant overall difference in cardiac mortality between Group I versus Group II patients (0 versus 1 patient, respectively, P=0.57). Intraoperative hypotension occurred in 4 patients in Group I versus 0 patient in Group II (P=0.03). Length of stay, graft function, and postoperative renal function were similar in both groups. CONCLUSION: A significant LVOT Delta >36 mmHg is a frequent finding occurring in 46/106 (43%) of patients who have DSE pre-OLT. Intraoperative hypotension is associated with patients having an LVOT Delta. However, post-OLT patients with significant LVOT Delta have a similar in-hospital outcome compared to patients without significant LVOT Delta. PMID- 15546369 TI - Electromechanical coupling, uncoupling, and ventricular function in patients with bundle branch block: a tissue-Doppler echocardiographic study. AB - BACKGROUND: Left bundle branch block (LBBB) is associated with impaired left ventricular (LV) function and increased morbidity and mortality, especially in patients with structural heart diseases. The mechanisms are poorly understood. SUBJECTS AND METHODS: Subjects with isolated LBBB (n=20), right bundle branch block (RBBB, n=20), and controls (C, n=20) were studied with standard two dimensional (2D), and color-encoded tissue-Doppler echocardiography (TDE). Inter- and intraventricular systolic and diastolic coordination were assessed from the TDE velocity profiles. LV function was assessed by 2D echocardiography, by TDE derived peak systolic velocities, and the atrioventricular (AV) plane displacement. RESULTS: Subjects with LBBB had longer electromechanical delays and longer isovolumic relaxation times than did the C and RBBB groups (P <0.001). For the LBBB subjects compared with the RBBB and C groups, ejection times were shorter, peak systolic velocities and AV plane displacements were lower, they had larger LV end-systolic volumes and lower LV ejection fraction (all P <0.001), and the atrial contribution to A-V plane displacement was higher (P <0.01). There were no differences in diastolic or filling times among the groups. CONCLUSIONS: In patients with LBBB, delayed regional electromechanical coupling and uncoupling leads to generalized intra- and interventricular asynchrony, thereby explaining the depressed regional and global LV functions. Assessment of the electromechanical coupling and uncoupling processes and their consequences on cardiac function in patients with BBB and structural heart diseases may be possible using TDE. PMID- 15546370 TI - Live three-dimensional transthoracic echocardiographic assessment of left ventricular hydatid cyst. AB - We report an adult patient in whom live three-dimensional transthoracic echocardiography (3DTTE) complemented two-dimensional transthoracic echocardiography (2DTTE) in making a definitive diagnosis of a hydatid cyst located in the left ventricular cavity. The parent hydatid cyst, as well as the daughter cysts, contained within it could be delineated by both 2DTTE and live 3DTTE. However, the tertiary or granddaughter cysts originating from the daughter cysts as well as great-granddaughter cysts budding from tertiary cysts could be visualized only when the live 3DTTE data sets were cropped and sectioned sequentially using multiple cutting planes. In addition, apparent intrinsic mobility of some of the tertiary cysts implying viability was detected only by 3DTTE. PMID- 15546371 TI - Usefulness of live three-dimensional transthoracic echocardiography in the characterization of atrial septal defects in adults. AB - In this report we present 12 adult patients in whom surgical or percutaneous intervention was considered for repair of atrial septal defect (ASD). Location, size, and surrounding atrial anatomy of the ASD were assessed prior to intervention in all patients with standard and live three-dimensional transthoracic echocardiography (3D TTE). In the four patients in whom intraoperative three-dimensional transesophageal echocardiographic reconstruction (3D TEE) was done, 3D TTE measurements of maximum dimension, maximum circumference, and maximum area of ASD agreed well with 3D TEE. In the seven patients who underwent transcatheter closure device insertion, live 3D TTE measurements of maximum dimension, maximum circumference, and maximum area of ASD agreed well with the sizing balloon. Additionally, since the sizing balloon measures a stretched diameter and area, a live 3D TTE stretched ASD diameter and area (derived from the actual live 3D TTE maximum circumference) were calculated and demonstrated improved agreement with the sizing balloon measurements. In all patients, > or =5 mm of atrial tissue was visualized surrounding the ASD. Further, with the addition of contrast enhancement, characterization of a small patent foramen ovale (<5 mm) was possible in one patient. Live 3D TTE accurately defined ASD location, size, and surrounding atrial anatomy in all patients studied by us. ASD characterization by live 3D TTE agreed well with 3D TEE and sizing balloon measurements. PMID- 15546372 TI - Atypical echocardiographic findings of endocarditis in an immunocompromised patient. AB - Cardiovascular manifestations in patients infected with human immunodeficiency virus (HIV) have been altered by the introduction of highly active antiretroviral therapy regimens that allow more effective prophylactic treatment and an increased time of survival. Because of this, noninfectious cardiac conditions associated with HIV disease are being recognized with increasing frequency in these patients. Cardiac involvement in HIV-infected patients varies from clinically silent to overtly symptomatic disease. By some estimates a direct cardiac cause of mortality is between 1% and 6% of all cases. Pericardial effusion, pericarditis, myocarditis, cardiomyopathy, endocarditis, and pulmonary hypertension are well-recognized cardiac illnesses associated with HIV infection. Echocardiography has been crucial in evaluating HIV-infected patients to assess the extent of cardiac involvement. This case report illustrates atypical echocardiographic manifestations of endocarditis and paravalvular abscess in an immunocompromised patient. PMID- 15546373 TI - Transesophageal echocardiography-guided transvenous biopsy of an intracardiac tumor. AB - Transesophageal echocardiography with simultaneous fluoroscopic guidance during a transvenous biopsy of an intracardiac tumor is a valuable technique. We present the case of a 67-year-old man with a metastasis in the right atrium from which histological examination of the tumor was indicated. The inaccessible location of the primary tumor in the liver and impaired hemostasis due to cirrhosis necessitated a transvenous biopsy of the metastasis. The procedure and the associated techniques are discussed in this case. PMID- 15546374 TI - Methodological analysis of diagnostic dobutamine stress echocardiography studies. AB - BACKGROUND: Dobutamine stress echocardiography (DSE) is an accepted test for the diagnosis of coronary artery disease (CAD), despite its wide diagnostic accuracy. AIM: Which factors cause test variability of DSE for the diagnosis of CAD. METHODS: In a retrospective analysis of 46 studies in 5,353 patients, the potential causes of diagnostic variability were systematically analyzed, including patient selection, definition of CAD, chest pain characteristics, confounding factors for DSE (left ventricular hypertrophy, left bundle branch block, female gender), work-up bias (present when patient's chance to undergo coronary angiography is influenced by the result of DSE), review bias (present when DSE is interpreted in relation to CAG), DSE protocol and definition of a positive DSE. RESULTS: Diagnostic variability was related to definition of a positive test, but not related to the definition of CAD or DSE protocol. However, only three of eight methodological standards for research design found general compliance. Differences in the selection of the study population (quality of echocardiographic window, angina pectoris), handling of confounding factors and analysis of disease in individual coronary arteries were observed. Lack of data on analysis of relevant chest pain syndromes and handling of nondiagnostic test results hampered further evaluation of these standards. CONCLUSION: Methodological problems may explain the wide range in diagnostic variability of DSE. An improvement of clinical relevance of DSE testing is possible by stronger adherence to common and new methodological standards. PMID- 15546375 TI - Quantification of mitral regurgitation by live three-dimensional transthoracic echocardiographic measurements of vena contracta area. AB - We evaluated 44 consecutive patients who underwent standard two-dimensional (2D) and live three-dimensional (3D) transthoracic echocardiography (TTE), as well as left heart catheterization with left ventriculography. Mitral regurgitant vena contracta area (VCA) was obtained by 3D TTE by systematic and sequential cropping of the acquired 3D TTE data set. Assessment of mitral regurgitation (MR) by ventriculography was compared to measurements of VCA by 3D TTE and to 2D TTE measurements of MR jet area to left atrial area (RJA/LAA), RJA alone, vena contracta width (VCW), and calculated VCA. VCA from 3D TTE closely correlated with angiographic grading (rs=0.88) with very little overlap. VCA of <0.2 cm2 correlated with mild MR, 0.2-0.4 cm2 with moderate MR, and >0.4 cm2 with severe MR by angiography. Ventriculographic grading also correlated well with 2D TTE measurements of RJA/LAA (rs=0.79) and RJA alone (rs=0.76) but with more overlap. Assessment of VCW and calculated VCA by 2D TTE agreed least with ventriculography (rs=0.51 and rs=0.55, respectively). Live 3D TTE color Doppler measurements of VCA can be used for quantitative assessment of MR and is comparable to assessment by ventriculography. PMID- 15546376 TI - Live three-dimensional transthoracic echocardiographic delineation of ventricular septal rupture following myocardial infarction. AB - We present an elderly patient with ventricular septal rupture following myocardial infarction in whom live three-dimensional transthoracic echocardiography allowed comprehensive noninvasive assessment of the location, shape, and size of the septal defect, which could be clearly visualized en face from both left and right ventricular aspects. PMID- 15546377 TI - Live three-dimensional transthoracic echocardiographic assessment of transcatheter closure of atrial septal defect and patent foramen ovale. AB - We report the usefulness of live three-dimensional transthoracic echocardiography in the accurate assessment of the morphology and efficacy of Amplatzer transcatheter devices used for closure of atrial septal defect and patent foramen ovale. PMID- 15546378 TI - Echo-color Doppler diagnosis of an unusual cause of neck mass. PMID- 15546379 TI - Visualization of the entry site of a dissection in the descending thoracic aorta by transthoracic echocardiography. PMID- 15546380 TI - Spectral Doppler instrumentation. PMID- 15546381 TI - Examination protocol for three-dimensional echocardiography. PMID- 15546382 TI - Cytokines and cytokine receptors. PMID- 15546383 TI - Interferons, interferon-like cytokines, and their receptors. AB - Recombinant interferon-alpha (IFN-alpha) was approved by regulatory agencies in many countries in 1986. As the first biotherapeutic approved, IFN-alpha paved the way for the development of many other cytokines and growth factors. Nevertheless, understanding the functions of the multitude of human IFNs and IFN-like cytokines has just touched the surface. This review summarizes the history of the purification of human IFNs and the key aspects of our current state of knowledge of human IFN genes, proteins, and receptors. All the known IFNs and IFN-like cytokines are described [IFN-alpha, IFN-beta, IFN-epsilon, IFN-kappa, IFN-omega, IFN-delta, IFN-tau, IFN-gamma, limitin, interleukin-28A (IL-28A), IL-28B, and IL 29] as well as their receptors and signal transduction pathways. The biological activities and clinical applications of the proteins are discussed. An extensive section on the evolution of these molecules provides some new insights into the development of these proteins as major elements of innate immunity. The overall structure of the IFNs is put into perspective in relation to their receptors and functions. PMID- 15546384 TI - The role of type I interferons in non-viral infections. AB - For a long time, the family of type I interferons (IFN-alpha/beta) has received little attention outside the fields of virology and tumor immunology. In recent years, IFN-alpha/beta regained the interest of immunologists, due to the phenotypic and functional characterization of IFN-alpha/beta-producing cells, the definition of novel immunomodulatory functions and signaling pathways of IFN alpha/beta, and the observation that IFN-alpha/beta not only exerts antiviral effects but is also relevant for the pathogenesis or control of certain bacterial and protozoan infections. This review summarizes the current knowledge on the production and function of IFN-alpha/beta during non-viral infections in vitro and in vivo. PMID- 15546385 TI - Lymphotoxin and LIGHT signaling pathways and target genes. AB - Lymphotoxins (LT alpha and LT beta), LIGHT [homologous to LT, inducible expression, competes with herpes simplex virus (HSV) glycoprotein D for HSV entry mediator (HVEM), a receptor expressed on T lymphocytes], tumor necrosis factor (TNF), and their specific receptors LT beta R, HVEM, and TNF receptor 1 (TNFR1) and TNFR2, form the immediate family of the larger TNF superfamily. These cytokines establish a critical communication system required for the development of secondary lymphoid tissues; however, knowledge of the target genes activated by these signaling pathways is limited. Target genes regulated by the LT alpha beta-LT beta R pathway include the tissue-organizing chemokines, CXCL13, CCL19, and CCL21, which establish cytokine circuits that regulate LT expression on lymphocytes, leading to organized lymphoid tissue. Infectious disease models have revealed that LT alpha beta pathways are also important for innate and adaptive immune responses involved in host defense. Here, regulation of interferon-beta by LT beta R and TNFR signaling may play a crucial role in certain viral infections. Regulation of autoimmune regulator in the thymus via LT beta R implicates LT/LIGHT involvement in central tolerance. Dysregulated expression of LIGHT overrides peripheral tolerance leading to T-cell-driven autoimmune disease. Blockade of TNF/LT/LIGHT pathways as an intervention in controlling autoimmune diseases is attractive, but such therapy may have risks. Thus, identifying and understanding the target genes may offer an opportunity to fine-tune inhibitory interventions. PMID- 15546386 TI - Cytokines and immunodeficiency diseases: critical roles of the gamma(c)-dependent cytokines interleukins 2, 4, 7, 9, 15, and 21, and their signaling pathways. AB - In this review, we discuss the role of cytokines and their signaling pathways in immunodeficiency. We focus primarily on severe combined immunodeficiency (SCID) diseases as the most severe forms of primary immunodeficiencies, reviewing the different genetic causes of these diseases. We focus in particular on the range of forms of SCID that result from defects in cytokine-signaling pathways. The most common form of SCID, X-linked SCID, results from mutations in the common cytokine receptor gamma-chain, which is shared by the receptors for interleukin (IL)-2, IL-4, IL-7, IL-9, IL-15, and IL-21, underscoring that X-linked SCID is indeed a disease of defective cytokine signaling. We also review the signaling pathways used by these cytokines and the phenotypes in humans and mice with defects in the cytokines or signaling pathways. We also briefly discuss other cytokines, such as interferon-gamma and IL-12, where mutations in the ligand or receptor or signaling components also cause clinical disease in humans. PMID- 15546387 TI - Biology of IL-21 and the IL-21 receptor. AB - Interleukin-21 (IL-21) is the newest member of the common gamma-chain family of cytokines, which includes IL-2, IL-4, IL-7, IL-9, IL-13, and IL-15. Its private receptor, IL-21R, has been shown to activate the Janus kinase/signal transducers and activators of transcription signaling pathway upon ligand binding. Initial studies have demonstrated that IL-21 has pleiotropic effects on the proliferation, differentiation, and effector functions of B, T, natural killer, and dendritic cells. More recently, the potential therapeutic capacity of IL-21 in the treatment of cancers has been widely investigated. The biological role of IL-21 in the immune system is complex, as IL-21 has been shown to have the ability to both promote and inhibit immune responses. Overall, the current data point to IL-21 being a novel immunomodulatory cytokine, whose regulation of any given immune response is highly dependent on the surrounding environmental context. PMID- 15546388 TI - IL-12 and IL-23: master regulators of innate and adaptive immunity. AB - Initiation of an effective immune response requires close interactions between innate and adaptive immunity. Recent advances in the field of cytokine biology have led to an increased understanding of how myeloid cell-derived factors regulate the immune system to protect the host from infections and prevent tumor development. In this review, we focus on the function of interleukin (IL)-23, a new member of the IL-12 family of regulatory cytokines produced by activated macrophages and dendritic cells. We propose that IL-12 and IL-23 promote two distinct immunological pathways that have separate but complementary functions. IL-12 is required for antimicrobial responses to intracellular pathogens, whereas IL-23 is likely to be important for the recruitment and activation of a range of inflammatory cells that is required for the induction of chronic inflammation and granuloma formation. These two cytokines work in concert to regulate cellular immune responses critical for host defense and tumor suppression. PMID- 15546389 TI - The role of IL-27 in the development of T-cell responses during parasitic infections. AB - The recognition that CD4+ T-cell responses could be divided into at least two functional subsets either dominated by production of interferon (IFN)-gamma and associated with cell-mediated immunity (Th1) or characterized by production of interleukin (IL)-4 and IL-5 and associated with humoral immunity (Th2) provided a basis to understand the role of T cells in resistance or susceptibility to different types of pathogens. As a consequence, many studies have focused on the identification of cytokines that influence these events. For example, the development of Th1-type responses is largely dependent on IL-12. However, other cytokines also affect this process, and initial studies revealed that IL-27, a cytokine with close structural and functional similarity to IL-12, can promote Th1 responses required for immunity to Leishmania major. Subsequent work with IL 27R (WSX-1)-deficient mice infected with Toxoplasma gondii or Trypanosoma cruzi revealed that the IL-27/IL-27R system can act as a negative regulator of inflammatory T-cell responses. The aim of this review is to discuss recent studies from these laboratories on the role of IL-27 in immunity to parasitic infections in the context of previous work and to highlight the pleiotropic effects of the IL-27/IL-27R system in the development and regulation of Th1 and Th2 responses. PMID- 15546390 TI - Induction of allergic inflammation by interleukin-18 in experimental animal models. AB - Interleukin-18 (IL-18) has been regarded as a proinflammatory cytokine because of its potent interferon-gamma-inducing activity. However, mutant mice that release excess amounts of IL-18 spontaneously develop pruritic chronic dermatitis-like atopic dermatitis (AD), suggesting the importance of IL-18 for the development of AD. Intriguingly, depletion of il-18 but not stat6, an essential transcriptional factor for IL-4 signaling, rescues the mice from AD, indicating IL-18-dependent, T-helper 2 (Th2) cell-independent AD. This type of AD is classified as innate type allergy in contrast to Th2 cell-dependent ordinary allergy. Consistent with the previous studies, mice transferred with antigen-specific Th1 cells exhibit no airway hyperresponsiveness and respiratory eosinophilic inflammation after challenge with antigen alone. However, they suffer from asthma upon challenge with antigen plus IL-18, with comparable levels of both the alterations as in those transferred with Th2 cells following challenge with antigen. The former type of asthma is categorized as Th1-associated allergy. Therefore, it is definitely necessary to evaluate whether individual allergic disorders involve either of these IL-18-mediated pathways or a Th2-mediated one. PMID- 15546391 TI - Signaling by IL-12 and IL-23 and the immunoregulatory roles of STAT4. AB - Produced in response to a variety of pathogenic organisms, interleukin (IL)-12 and IL-23 are key immunoregulatory cytokines that coordinate innate and adaptive immune responses. These dimeric cytokines share a subunit, designated p40, and bind to a common receptor chain, IL-12R beta 1. The receptor for IL-12 is composed of IL-12R beta 1 and IL-12R beta 2, whereas IL-23 binds to a receptor composed of IL-12R beta 1 and IL-23R. Both cytokines activate the Janus kinases Tyk2 and Jak2, the transcription factor signal transducer and activator of transcription 4 (STAT4), as well as other STATs. A major action of IL-12 is to promote the differentiation of naive CD4+ T cells into T-helper (Th) 1 cells, which produce interferon (IFN)-gamma, and deficiency of IL-12, IL-12R subunits or STAT4 is similar in many respects. In contrast, IL-23 promotes end-stage inflammation. Targeting IL-12, IL-23, and their downstream signaling elements would therefore be logical strategies for the treatment of immune-mediated diseases. PMID- 15546392 TI - Issues in T-helper 1 development--resolved and unresolved. AB - T-helper 1 cell (Th1) development participates in immunity to many pathogens in part by providing a source of interferon (IFN)-gamma that contributes numerous protective effects. The process of Th1 development involves signals provided by antigen-presenting cells and cytokines produced in response to pathogens, with IFN-gamma itself, interleukin (IL)-12, and IL-18 each promoting the process in some way. Despite the rapid progress into mechanisms of Th1 development in recent years, there are still a number of important unresolved issues in this area. The precise sequence of effector and cellular mechanisms represents a relatively recent avenue of research but is still the subject of current debate, as is the basis of mechanisms that may stabilize a Th1 response. Another unresolved issue is the role of type I IFNs in substituting for IL-12-mediated activation of signal transducer and activator of transcription 4 (Stat4) and induction of IFN gamma in either murine or human T cells. It is now clear that Th1 cells acquire the property of being capable of nonantigen-dependent activation through the coordinate signaling of IL-12 and IL-18, but the precise order of intracellular signaling events and the uniqueness of this pathway's reliance on the p38 mitogen activated protein kinase (MAPK) pathway are still issues in need of resolution. Finally, the process of verifying the effects of Stat4 mutations on functional responses has led to the recognition of an unexpected action of the STAT N-domain that may apply generally to other STAT proteins as well. None of these areas is static or resolved fully, and they likely will remain topics of rapid progress. PMID- 15546393 TI - Interleukin-13 in asthma pathogenesis. AB - Bronchial asthma is a complex disorder that is thought to arise as a result of aberrant T-lymphocyte responses to noninfectious environmental antigens. In particular, the symptoms of asthma are closely associated with the presence of activated T-helper 2 cell (Th2) cytokine-producing cells [interleukin (IL)-4, IL 5, IL-9, and IL-13] in the airway wall. Although each of the Th2 cytokines likely contributes to the overall immune response directed against environmental antigens, a substantial body of evidence points to a singular role for IL-13 in the regulation of the allergic diathesis. Initial studies in animal models of disease provided compelling evidence that IL-13, independently of other Th2 cytokines, was both necessary and sufficient to induce all features of allergic asthma. The importance of IL-13 in allergic disorders in humans is supported by consistent associations between tissue IL-13 levels and genetic variants in the IL-13 gene with asthma and related traits. With the preponderance of evidence continuing to support a pivotal role for IL-13 in allergic disorders, attention is now turned toward understanding the mechanisms by which this cytokine may mediate the pathophysiological features of allergic disease. The emerging paradigm is that IL-13 induces features of the allergic response via a complex array of actions on resident airway cells rather than through traditional effector pathways involving eosinophils and immunoglobulin E-mediated events. In light of these recent developments, this review explores our current understanding of the singular role of IL-13 in the pathogenesis of asthma, with a particular focus on new insights into the mechanisms by which IL-13 mediates various features of asthma. PMID- 15546394 TI - Opposing roles for IL-13 and IL-13 receptor alpha 2 in health and disease. AB - Interleukin (IL)-13 is a key inducer of several type-2 cytokine-dependent pathologies. It regulates inflammation, mucus production, tissue remodeling, and fibrosis. Consequently, it has become an important therapeutic target for a number of debilitating illnesses, including asthma, idiopathic pulmonary fibrosis, ulcerative colitis, as well as several other diseases in which IL-13 is believed to be overproduced. In the murine model of schistosomiasis, IL-13 has emerged as a central mediator of chronic infection-induced liver pathology. Although IL-4, IL-5, IL-10, and IL-13 each regulate distinct aspects of the granulomatous inflammatory response, IL-13 was identified as the primary mediator of liver fibrosis. Thus, elucidating the mechanisms that regulate the production and function of IL-13 has become an intensive area of research. IL-13 signaling is mediated by the type-2 IL-4 receptor, which consists of the IL-4R alpha and IL 13R alpha 1 chains. However, another IL-13-binding chain, IL-13R alpha 2, appears to strongly inhibit the activity of IL-13. Animals deficient in IL-13R alpha 2 fail to downmodulate granuloma formation in the chronic phase of infection. They also develop severe IL-13-dependent fibrosis and portal hypertension and quickly succumb to the infection. Here, we summarize findings from the schistosomiasis model, which illustrate opposing activities for IL-13 and IL-13R alpha 2 in health and disease. PMID- 15546395 TI - Signaling pathways in Th2 development. AB - In order for an immune response to be successful, it must be of the appropriate type and magnitude. Intracellular residing pathogens require a cell-mediated immune response, whereas extracellular pathogens evoke a humoral immune response. T-helper (Th) cells orchestrate the immune response and are divided into two subsets, Th1 and Th2 cells. Here, we discuss the mechanisms of Th2 development with a focus on signal transduction pathways that influence Th2 differentiation. PMID- 15546396 TI - Interleukin-10 in viral diseases and cancer: exiting the labyrinth? AB - Interleukin-10 (IL-10) is unique among cytokines, as it is considered both as a potent immunostimulatory and immunosuppressive factor. This complex biology has been particularly challenging when trying to define the useful or harmful role of IL-10 in chronic viral diseases and cancer. In the present review, we emphasize how these multiple roles define IL-10 as an adaptive molecule, constantly tuning the host response against dangerous and resourceful pathogens. PMID- 15546397 TI - The BLyS family of ligands and receptors: an archetype for niche-specific homeostatic regulation. AB - Discovery and characterization of the tumor necrosis factor (TNF) family member B lymphocyte stimulator (BLyS) has opened a novel chapter in the role of TNF family members in the homeostatic control of lymphocyte populations. BLyS and its sister cytokine APRIL (a proliferation-inducing ligand) act primarily as soluble trimers and serve to regulate the steady-state numbers of nearly all B-cell compartments. This homeostatic regulation is accomplished through the regulation of B-cell production rates, selection thresholds, and lifespan. Differential expression of the three BLyS receptors during differentiation and activation provides related yet distinct homeostatic niches for follicular, marginal zone, and memory B-cell subsets. PMID- 15546398 TI - Is targeting Toll-like receptors and their signaling pathway a useful therapeutic approach to modulating cytokine-driven inflammation? AB - Cytokine-driven inflammation and tissue destruction is a common theme of chronic inflammatory diseases such as rheumatoid arthritis, Crohn's disease, ulcerative colitis, psoriasis, chronic obstructive pulmonary disease, and atherosclerosis. Research over the last two decades demonstrated the importance of cytokines that are not only expressed chronically but also are capable of signaling at sites of chronic inflammation. Cytokines thus regulate major pathological processes that include inflammation, angiogenesis, tissue remodeling, and fibrosis. This research led to the identification of key cytokines involved in these processes, two of which, tumor necrosis factor-alpha and interleukin-1, have also been successfully targeted in the clinic. However, what triggers and maintains cytokine gene expression in chronic inflammation remains a mystery. In this article, we review current progress in the understanding of cytokine-driven inflammation and discuss current evidence implicating Toll-like receptors (TLRs), recently identified as the receptors recognizing self versus non-self molecular patterns, in the regulation of cytokine-driven inflammation. Whether targeting TLRs and their downstream signaling pathway will prove to be a successful approach for the treatment of these devastating diseases remains to be determined. PMID- 15546399 TI - The role of cytokine DNAs as vaccine adjuvants for optimizing cellular immune responses. AB - Cytokines represent a diverse group of immunologic effector and regulatory proteins that are critical components of the host response to invading pathogens. They have also been utilized as adjuvants to enhance immune responses to vaccines. In particular, plasmid cytokines have been studied extensively as candidate adjuvants for DNA vaccines in preclinical models and are now entering early-phase clinical trials. Here, we review recent advances in our understanding of cytokine biology, T-lymphocyte differentiation, and potential applications of plasmid cytokines in the rational design of improved vaccines. PMID- 15546400 TI - Cytokines in cancer immunity and immunotherapy. AB - The concept that the immune system recognizes and controls cancer was first postulated over a century ago, and cancer immunity has continued to be vigorously debated and experimentally tested. Mounting evidence in humans and mice supports the involvement of cytokines in tumor initiation, growth, and metastasis. The idea that the immune system detects stressed, transformed, and frankly malignant cells underpins much of the excitement currently surrounding new cytokine therapies in cancer treatment. In this review, we define the contrasting roles that cytokines play in promoting tumor immunity, inflammation, and carcinogenesis. We also discuss the more promising aspects of clinical cytokine use in cancer patients. PMID- 15546401 TI - Direct detection of bacterial pathogens in representative dairy products using a combined bacterial concentration-PCR approach. AB - AIMS: To develop a simple, rapid method to concentrate and purify bacteria and their nucleic acids from complex dairy food matrices in preparation for direct pathogen detection using polymerase chain reaction (PCR). METHODS AND RESULTS: Plain non-fat yogurt and cheddar cheese were each seeded with Listeria monocytogenes or Salmonella enterica serovar. Enteritidis in the range of 10(1) 10(6) CFU per 11-g sample. Samples were then processed for bacterial concentration using high-speed centrifugation (9700 g) followed by DNA extraction, PCR amplification, and amplicon confirmation by hybridization. Bacterial recoveries after centrifugation ranged from 53 to >100% and 71 to >100% for serovar. Enteritidis and L. monocytogenes, respectively, in the non-fat yogurt samples; and from 77 to >100% and 69 to >100% for serovar. Enteritidis and L. monocytogenes, respectively, in the cheddar cheese samples. There were no significant differences in recovery efficiency at different inocula levels, and losses to discarded supernatants were always <5%, regardless of dairy product or pathogen. CONCLUSIONS: When followed by pathogen detection using PCR and confirmation by amplicon hybridization, detection limits of 10(3) and 10(1) CFU per 11-g sample were achieved for L. monocytogenes and serovar. Enteritidis, respectively, in both product types and without prior cultural enrichment. SIGNIFICANCE AND IMPACT OF THE STUDY: This study represents progress toward the rapid and efficient direct detection of pathogens from complex food matrices at detection limits approaching those that might be anticipated in naturally contaminated products. PMID- 15546402 TI - Bacterial population dynamics and community structure in a pharmaceutical manufacturing water supply system determined by real-time PCR and PCR-denaturing gradient gel electrophoresis. AB - AIMS: To control bacteria in the pharmaceutical water supply system. METHODS AND RESULTS: Bacteria were enumerated by conventional culture method and fluorescent vital staining. Activated carbon treatment and storage in a tank provided favourable environments for bacterial growth. The bacterial population of the water in both the post-activated carbon treatment and the tank was analysed by denaturing gradient gel electrophoresis (DGGE) with PCR-amplified 16S rDNA fragments including V6, -7, and -8 regions. The bacterial community structure in activated carbon treated water was stable throughout the year. Several kinds of bacteria such as genus Aquaspirillum and Methylobacterium were found in the water after activated carbon treatment. The bacterial community structure was changed and other bacteria such as mycobacteria were detected after storage. Mycobacteria were quantified in water samples using real-time PCR targeting the 16S rDNA gene. Mycobacteria were also detected in tap water and their number was increased 10(3) 10(4)-fold higher after storage. CONCLUSION: These data suggest the importance of culture-independent methods for quality control of water used in pharmaceutical manufacturing. SIGNIFICANCE AND IMPACT OF THE STUDY: Critical steps and specified bacteria that should be controlled in the water supply system were recognized by culture-independent methods. These data will enable effective control of water used in the pharmaceutical industry. PMID- 15546403 TI - In situ quantification of biocide efficacy using GFP transformed Aureobasidium pullulans. AB - AIMS: To develop a real-time in situ method to quantify loss of viability of Aureobasidium pullulans PRAFS8 cells attached to plasticized polyvinyl chloride (pPVC) with incorporated biocides, and to use the method to compare biocide efficacy in situ. METHODS AND RESULTS: A. pullulans PRAFS8, transformed with green fluorescent protein (GFP), was used to quantify the efficacy of a range of biocides incorporated into pPVC. Experimentally, it was found that a density of 1.53 x 10(6) yeast cells per cm(2) of pPVC was optimal as increasing the density of the yeast cells to 6.12 x 10(6) cm(-2) attached to pPVC containing the biocide 2-n-octyl-4-isothiazolin-3-one (OIT) decreased the rate of fluorescence loss. A strong positive correlation between fluorescence and viable yeast cell number was observed and fluorescence was used as a direct indicator of cell viability. The effectiveness of five commercial biocides, commonly incorporated into pPVC at their in-use concentrations, was tested against yeast cells attached to the pPVC surface. The loss of fluorescence and hence viability in situ was quantified using image analysis. The biocides N-(trichloromethylthio) phthalimide (NCMP), 10,10'-oxybisphenoxarsine (OBPA), OIT and 2,3,5,6-tetrachloro-4-(methylsulphonyl) pyridine (TCMP) caused complete loss of fluorescence within 30-50 h. In contrast the biocide dichloro-octyl-isothiazoline caused only 55 +/- 15% fluorescence loss after 50 h. Starvation of the yeast cells in suspension for 24 h prior to attachment reduced their initial sensitivity to OBPA, NCMP, OIT and TCMP by 15 20%, but eventually the fluorescence was also completely lost. CONCLUSIONS: The use of A. pullulans expressing cytosolic GFP enables the in situ quantification of loss of viability when cells are attached to pPVC with incorporated biocides. SIGNIFICANCE AND IMPACT OF THE STUDY: GFP fluorescence was used as a real-time indicator of cell viability and thus can be applied for direct quantification of the effectiveness of a broad range of biocides, incorporated into the polymer mass and used to protect a variety of plastics or other materials from microbial growth. PMID- 15546404 TI - Predominance of Saccharomyces uvarum during spontaneous alcoholic fermentation, for three consecutive years, in an Alsatian winery. AB - AIMS: The purpose of this study was to determine the origin of the yeasts involved in the spontaneous alcoholic fermentation of an Alsatian wine. METHODS AND RESULTS: During three successive years, must was collected at different stages of the winemaking process and fermented in the laboratory or in the cellar. Saccharomyces yeasts were sampled at the beginning and at the end of the fermentations. Saccharomyces cerevisiae clones were genetically characterized by inter-delta PCR. Non-S. cerevisiae clones were identified as Saccharomyces uvarum by PCR-RFLP on MET2 gene and characterized at the strain level by karyotyping. The composition of the Saccharomyces population in the vineyard, after crushing and in the vat was analyzed. This led to three main results. First, the vineyard Saccharomyces population was rather homogeneous. Second, new non-resident strains had appeared in the must during the winemaking process. Finally, the yeast population in the vat only consisted in S. uvarum strains. CONCLUSION: This 3 year study has enabled us to show the involvement of indigenous S. uvarum in the alcoholic fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study gives a first insight into the polymorphism of S. uvarum strains involved in a spontaneous alcoholic fermentation. PMID- 15546405 TI - Use of the 'ex vivo' test to study long-term bacterial survival on human skin and their sensitivity to antisepsis. AB - AIMS: To determine bacterial survival on human skin and their sensitivity to antisepsis. METHODS AND RESULTS: An 'ex vivo' protocol which uses human skin samples placed into diffusion cells, and electron microscopy (EM), were used to study the growth of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa inoculated onto skin samples over a 46-h incubation period at 32 degrees C. Concurrently variation in skin pH was evaluated at different time intervals during this period. In addition the antimicrobial activity of three antiseptics against the incubated micro-organisms was assessed quantitatively with the 'ex vivo' test, while their detrimental effects against bacteria were observed by EM. All three bacteria were still present in high number after 46 h inoculation on skin, although the concentration of E. coli and S. aureus were reduced by 2.74 and 1.58 log(10) reduction, respectively, over this period of time. Electron micrographs showed clear evidence of cell division and some bacteria appeared to be embedded into the skin layers. The antiseptics tested had some antibacterial activity against bacteria incubated on skin for 3 and 10 h, and EM evidence showed some morphological damages including cellular blebbing and the presence of fibrillar material around the cells. All micro-organisms had an acidifying effect on skin samples. CONCLUSIONS: Here, it was shown that bacterial pathogens can survive and grow when incubated on human skin. In addition, it is possible that they can penetrate the stratum corneum, which can provide some protection against antisepsis. SIGNIFICANCE AND IMPACT OF THE STUDY: The apparent low bactericidal activity of biocides attributed in part to bacterial protection from skin layers is particularly important to assess in order to ensure antisepsis efficacy. PMID- 15546406 TI - Development of a capture/enrichment sandwich ELISA for the rapid detection of enteropathogenic and enterohaemorrhagic Escherichia coli O26 strains. AB - AIMS: To improve the sensitivity of a monoclonal antibody (MAb 2F3) based enteropathogenic Escherichia coli (EPEC)/enterohaemorrhagic E. coli (EHEC) serogroup O26-specific sandwich ELISA (sELISA), using a capture/enrichment format of the assay. METHODS AND RESULTS: The sELISA utilized an EPEC/EHEC O26-specific MAb 2F3 as the capture reagent and an E. coli serogroup O26 lipopolysaccharide specific polyclonal antibody in the development stage. Wells containing faeces test samples from bovine enteritis cases and agar colony sweep cultures from human diarrhoea cases, after a 2-h capture stage, were washed and enrichment of the captured cells was encouraged by addition of tryptone soya broth. After overnight incubation, the contents of each well were transferred to sterile wells and the sELISA completed. Any sELISA positive samples were then subcultured onto blood agar to recover and further characterize the positive cultures. The assay had a sensitivity of 10(3) CFU ml(-1). ELISA positive samples consisted of 21 (4.8%) of the 442 bovine and 19 (3.7%) of the 519 human samples tested, and ELISA positive EPEC/EHEC O26 strains were isolated from 11 and three of these samples respectively. CONCLUSION: The capture/enrichment method improved the sensitivity of a MAb-based sELISA for the detection of EPEC/EHEC O26 strains, and also contributed to an improved isolation rate of the organism from field samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The application of a specific MAb in a capture/enrichment format of the sELISA, provides a prospectively suitable screening method for the detection of pathogenic bacteria from mixed culture samples. PMID- 15546407 TI - Development of an extensive set of 16S rDNA-targeted primers for quantification of pathogenic and indigenous bacteria in faecal samples by real-time PCR. AB - AIMS: The microbiota of the human intestinal tract constitutes a complex ecosystem. We report the design and optimization of an extensive set of 16S rDNA targeted species- and group-specific primers for more accurate quantification of bacteria from faecal samples with real-time PCR. METHODS AND RESULTS: A linear range of quantification between 0.1-10 pg and 10 ng of specific target genome was obtained, which corresponds to detection of ca 30-4500 to 1.9 x 10(6)-6.0 x 10(6) target bacterial genomes. Functionality of the assays was confirmed by quantification of target bacterial DNA from faecal DNA preparations of healthy volunteers and irritable bowel syndrome (IBS) patients. Additionally, spiking of faecal preparations with Helicobacter pylori, Clostridium difficile or Campylobacter jejuni was used to confirm the accurate and sensitive quantification. CONCLUSIONS: Real-time PCR is a very sensitive and precise technique for an extensive quantitative evaluation of gut microbiota and is feasible for detection of human pathogens from faecal samples. SIGNIFICANCE AND IMPACT OF THE STUDY: To design and optimize an extensive set of real-time PCR assays targeting a large group of predominant and pathogenic GI microbial species for further use in updating the current knowledge of the putative role of gut microbiota in health and disease. PMID- 15546408 TI - Deletion of the Delta12-oleic acid desaturase gene of a nonaflatoxigenic Aspergillus parasiticus field isolate affects conidiation and sclerotial development. AB - AIMS: To investigate how linoleic acid affects conidial production and sclerotial development in a strictly mitotic Aspergillus parasiticus field isolate as related to improving biocompetitivity of atoxigenic Aspergillus species. METHODS AND RESULTS: We disrupted A. parasiticusDelta12-oleic acid desaturase gene (odeA) responsible for the conversion of oleic acid to linoleic acid. We examined conidiation and sclerotial development of SRRC 2043 and three isogenic mutant strains deleted for the odeA gene (DeltaodeA), either with or without supplementing linoleic acid, on one complex potato dextrose agar (PDA) medium and on two defined media: nitrate-containing Czapek agar (CZ) and Cove's ammonium medium (CVN). The DeltaodeA mutants produced less conidia than the parental strain on all media. Linoleic acid supplementation (as sodium linoleate at 0.3 and 1.2 mg ml(-1)) restored the DeltaodeA conidial production comparable to or exceeding the unsupplemented parental level, and the effect was medium dependent, with the highest increase on CVN and the least on PDA. SRRC 2043 and the DeltaodeA mutants were unable to produce sclerotia on CVN. On unsupplemented PDA and CZ, DeltaodeA sclerotial mass was comparable to that of SRRC 2043, but sclerotial number increased significantly to two- to threefold. Supplementing linoleic acid to media, in general, tended to decrease wild type and DeltaodeA sclerotial mass and sclerotial number. CONCLUSIONS: Linoleic acid stimulates conidial production but has an inhibitory effect on sclerotial development. The relationship between the two processes in A. parasiticus is complex and affected by multiple factors, such as fatty acid composition and nitrogen source. SIGNIFICANCE AND IMPACT OF STUDY: Conditions that promote sclerotial development differ from those required to promote maximum conidial production. Manipulation of content and availability of linoleic acid at different fungal growth phases might optimize conidial and sclerotial production hence increasing the efficacy of biocompetitive Aspergillus species. PMID- 15546409 TI - Growth of enterotoxigenic Staphylococcus aureus in povi masima, a traditional Pacific island food. AB - AIMS: To obtain preliminary data on the microbiology and hurdles to pathogen growth in the traditional Pacific Island food, povi masima, which is essentially beef brisket cured in brine. METHODS AND RESULTS: Six containers of povi masima were prepared and two were inoculated with five enterotoxigenic strains of Staphyloccocus aureus. The povi masima were divided into two lots each containing two uninoculated control and an inoculated container. Lot 1 was incubated at room temperature (20 degrees C) and lot 2 under refrigeration (4-5 degrees C) for up to 98 days. During storage, samples were removed and tested for aerobic plate count, coagulase-producing Staphylococci, Clostridium perfringens, staphylococcal enterotoxin and various chemical parameters of the food. Coagulase-producing Staphylococci and aerobic plate counts grew to high levels in both the inoculated and uninoculated lots stored at room temperature, but enterotoxin was only detected at one time point in these lots and this may represent a false positive result. The concentration of NaCl in the meat increased with time as concentrations equilibrated, and nitrite was rapidly lost in those lots stored at room temperature. Storage at 4-5 degrees C prevented proliferation of coagulase producing Staphylococci. CONCLUSIONS: For safe curing and storage, this food should be kept under refrigeration as this prevented growth of staphylococci. Optimum storage would also be achieved with improved attempts to ensure equal distribution of NaCl prior to storage. SIGNIFICANCE AND IMPACT OF THE STUDY: Under conditions traditionally used to cure and store this food, enterotoxigenic staphylococci can grow to numbers where toxigenesis might occur, especially during the early stages of curing where the salt has not diffused from the brine into the meat. PMID- 15546410 TI - The role of host organism, transcriptional switches and reporter mechanisms in the performance of Hg-induced biosensors. AB - AIMS: The purpose of this study was to comprehensively compare the response of nine biosensors capable of being induced by Hg. Induction by Hg was based upon the insertion of merR, merB, zntA and zntR promoter genes. LuxCDABE or lucFF reporter genes expressed luminescence, and host organisms were Escherichia coli, Vibrio anguillarum and Pseudomonas fluorescens. The role of transcriptional switches, reporter mechanism and host organism was to be investigated. METHODS AND RESULTS: All biosensors were subjected to the same assay conditions. Sensors had their own individual growth characteristics and response to the doses of Hg tested. Maximum bioluminescence response was induced by concentrations of Hg between 2.5 nm and 5 microM. E. coli pRB28 was found to detect levels of Hg as low as 1.6 nm and yet was capable of operating in a concentration range of up to 12.5 microM. CONCLUSIONS: The response of the sensors demonstrated their suitability for analysis under environmentally relevant concentrations. The sensitivity of the sensors, the optimum range and the expediency of the assay could not be related to a single sensor trait. It may be concluded that biosensor performance is dependent on more than one of the single factors studied. SIGNIFICANCE AND IMPACT OF THE STUDY: The results show that comparative testing of sensors is an important step in evaluating the relevance and performance of biosensors prior to routine environmental application. PMID- 15546411 TI - Penicillium chrysogenum glucose oxidase -- a study on its antifungal effects. AB - AIMS: Purification and characterization of the high molecular mass Candida albicans-killing protein secreted by Penicillium chrysogenum. METHODS AND RESULTS: The protein was purified by a combination of ultrafiltration, chromatofocusing and gel filtration. Enzymological characteristics [relative molecular mass (M(r)) = 155 000, subunit structure alpha(2) with M(r,alpha) = 76 000, isoelectric point (pI) = 5.4] were determined using SDS-PAGE and 2D electrophoresis. N-terminal amino acid sequencing and homology search demonstrated that the antifungal protein was the glucose oxidase (GOX) of the fungus. The enzyme was cytotoxic for a series of bacteria, yeasts and filamentous fungi. Vitamin C (1.0 mg ml(-1)) prevented oxidative cell injuries triggered by 0.004 U GOX in Emericella nidulans cultures but bovine liver catalase was ineffective even at a GOX : catalase activity ratio of 0.004 : 200 U. A secondary inhibition of growth in E. nidulans cultures by the oxygen-depleting GOX-catalase system was likely to replace the primary inhibition exerted by H(2)O(2). CONCLUSIONS: Penicillium chrysogenum GOX possesses similar enzymological features to those described earlier for other Penicillium GOXs. Its cytotoxicity was dependent on the inherent antioxidant potential of the test micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: Penicillium chrysogenum GOX may find future applications in glucose biosensor production, the disinfection of medical implants or in the food industry as an antimicrobial and/or preservative agent. PMID- 15546412 TI - Molecular diversity of tannic acid degrading bacteria isolated from tannery soil. AB - AIMS: The aim of this study was to enrich and isolate bacteria from a tannery soil that were capable of utilizing tannic acid and gallic acid as sole source of carbon aerobically, and to characterize their diversity in order to identify efficient strains that can be used for tannin bioremediation. METHODS AND RESULTS: Bacterial strains were isolated after enrichment in minimal medium with tannic acid or gallic acid as sole carbon source. Polymerase chain reaction (PCR) restricted fragment length polymorphism of 16S rDNA [amplified ribosomal DNA restriction analysis (ARDRA)] and BOX-PCR was used to characterize their diversity. Two strains showing relatively high efficiency in degrading tannic acid and gallic acid were identified on the basis of carbon source utilization pattern (BIOLOG) and 16S rDNA sequence. CONCLUSIONS: Bacterial strains capable of degrading tannic acid and gallic acid could be grouped into six and seven clusters on the basis of ARDRA and BOX-PCR, respectively. On the basis of 16S rDNA sequence, the most efficient isolate degrading tannic acid belonged to Pseudomonas citronellolis, whereas the most efficient gallic acid degrader showed maximum phylogenetic relatedness to P. plecoglossicida. SIGNIFICANCE AND IMPACT OF THE STUDY: Aerobic tannic acid degraders such as the two strains isolated in this study can be used for tannin bioremediation, and in the study of genes involved in the production of tannase, an industrially important enzyme. PMID- 15546413 TI - Effect of microwave radiation on Bacillus subtilis spores. AB - AIMS: To compare the killing efficacy and the effects exerted by microwaves and conventional heating on structural and molecular components of Bacillus subtilis spores. METHODS AND RESULTS: A microwave waveguide applicator was developed to generate a uniform and measurable distribution of the microwave electric-field amplitude. The applicator enabled the killing efficacy exerted by microwaves on B. subtilis spores to be evaluated in comparison with conventional heating at the same temperature value. The two treatments produced a similar kinetics of spore survival, while remarkably different effects on spore structures were seen. The cortex layer of the spores subjected to conductive heating was 10 times wider than that of the untreated spores; in contrast, the cortex of irradiated spores did not change. In addition, the heated spores were found to release appreciable amounts of dipicolinic acid (DPA) upon treatment, while extracellular DPA was completely undetectable in supernatants of the irradiated spores. These observations suggest that microwave radiation may promote the formation of stable complexes between DPA and other spore components (i.e. calcium ions); thus, making any release of DPA from irradiated spores undetectable. Indeed, while a decrease in measurable DPA concentrations was not produced by microwave radiation on pure DPA solutions, a significant lowering in DPA concentration was detected when this molecule was exposed to microwaves in the presence of either calcium ions or spore suspensions. CONCLUSIONS: Microwaves are as effective as conductive heating in killing B. subtilis spores, but the microwave E-field induces changes in the structural and/or molecular components of spores that differ from those attributable only to heat. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides information on the effect of microwaves on B. subtilis spore components. PMID- 15546414 TI - Description of heterotrophic bacteria occurring in paper mills and paper products. AB - AIMS: To isolate aerobic mesophilic bacilli and thermophilic bacteria from different paper mill samples and to evaluate their potential harmfulness. METHODS AND RESULTS: A total of 109 mesophilic and 68 thermophilic isolates were purified and characterized by automated ribotyping and partial 16S rDNA sequencing. The mesophilic isolates belonged to the genera Bacillus (13 taxa), Brevibacillus (three taxa) and Paenibacillus (five taxa). The thermophilic bacteria represented seven taxa of Bacillus, Geobacillus or Paenibacillus, four of proteobacteria and one of actinobacteria. The most frequently occurring bacteria were Bacillus cereus, B. licheniformis, Pseudoxanthomonas taiwanensis and bacteria closely related to Paenibacillus stellifer, P. turicensis or Leptothrix sp. One mill was contaminated throughout with bacteria of a novel mesophilic genus most closely related to Brevibacillus centrosporus and another with bacteria of a novel thermophilic genus most closely related to Hydrogenophilus thermoluteolus. One B. cereus isolate producing haemolytic diarrhoeal enterotoxin was detected and all the tested B. licheniformis isolates produced a metabolite toxic to boar sperm cells. CONCLUSIONS: The bacilli and thermophilic bacteria isolated represent species which should not present occupational hazards in paper mill environments. The most harmful bacterium detected was B. licheniformis and potentially also B. cereus. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the microbial diversity in a paper mill provides a rational basis for development of an effective controlling programme. A database constructed from the fingerprints generated using automated ribotyping helps to identify and trace the contamination routes of bacteria occurring in paper mills. PMID- 15546415 TI - Inhibition of the reattachment of young adult zebra mussels by single-species biofilms and associated exopolymers. AB - AIMS: To determine the effects of single-species bacterial films and their associated extracellular products on the reattachment of young adult zebra mussels. MATERIALS AND RESULTS: Ten strains of bacteria were isolated from surfaces where adult zebra mussels can be found attached in nature. Single species biofilms were developed on both glass and polystyrene using these bacteria. The reattachment of zebra mussels (i.e. with byssal threads) was compared between surfaces with and without films. Although no differences were observed in mussel reattachment between glass surfaces with and without films (P > 0.05, anova), a reduction in mussel reattachment between polystyrene surfaces with and without films was observed for seven of the 10 strains (P < or = 0.05 to <0.001, anova). Bacterial extracellular products (BEP) were isolated from five bacterial films and tested for their effects on mussel reattachment. Four of the five sets of isolated extracellular products evoked the same effects as their respective intact biofilms. CONCLUSIONS: We conclude that depending on the substratum, individual strains of bacteria in biofilms can inhibit the reattachment of adult zebra mussels. In some cases, BEP were the source of the inhibitory effects. SIGNIFICANCE AND IMPACT OF THE STUDY: The nature of the substratum on which the biofilms develop affects properties of the biofilm and its extracellular components, which subsequently influences zebra mussel reattachment. PMID- 15546416 TI - Genetic and functional characterization of a Bacillus sp. strain excreting surfactin and antifungal metabolites partially identified as iturin-like compounds. AB - AIMS: A bacterial strain producing antifungal compounds active against the plant pathogenic fungi Fusarium, Rhizoctonia and Sclerotinia has been characterized and shown to control Rhizoctonia root rot of soya bean. METHODS AND RESULTS: The metabolites excreted by Bacillus BNM 122 remained active after autoclaving, were resistant over a wide pH range and to hydrolytic enzymes. By (1)H-NMR and thin layer chromatography analyses surfactin and iturin-like compounds were partially identified. Moreover, soya bean seeds bacterization with BNM 122 in a compost based formulation was as effective controlling Rhizoctonia solani as pentachloronitrobenzene. According to its 16S rDNA sequence BNM 122 was closely related to Bacillus amyloliquefaciens and Bacillus subtilis. PCR analysis of the 16S-23S rRNA intergenic spacer region and repetitive sequence-based PCR (rep-PCR) genomic fingerprinting revealed a close genetic relationship to B. amyloliquefaciens. However, by physiological characterization using API tests, this strain resembled more B. subtilis. CONCLUSIONS: This is the first report describing the co-production of surfactin and iturin-like compounds by a putative strain of B. amyloliquefaciens. The synergistic effect of both lipopetides is a remarkable trait for a candidate biocontrol agent. SIGNIFICANCE AND IMPACT OF THE STUDY: This kind of research has relevance in order to minimize the use of synthetic fungicides and surfactants, contributing to the preservation of the environment. PMID- 15546417 TI - Fermentation conditions affecting the bacterial growth and exopolysaccharide production by Streptococcus thermophilus ST 111 in milk-based medium. AB - AIMS: To study the effect of different fermentation conditions and to model the effect of temperature and pH on different biokinetic parameters of bacterial growth and exopolysaccharides (EPS) production of Streptococcus thermophilus ST 111 in milk-based medium. METHODS AND RESULTS: The influence of temperature and pH was studied through fermentation and modelling. Fermentations under non-pH controlled conditions with S. thermophilus ST 111 indicated that the EPS production was low in milk medium, even if additional nitrogen sources were supplemented. Under pH-controlled conditions, addition of whey protein hydrolysate to the milk medium resulted in a fivefold increase of the EPS production. This medium did not contain polysaccharides interfering with EPS isolation. Primary and secondary modelling of different fermentations revealed an optimum temperature and pH of 40 degrees C and constant pH 6.2, respectively, for growth in milk medium supplemented with whey protein hydrolysate. Maximum EPS production was observed in the range of 32-42 degrees C and constant pH 5.5-6.6. Whereas growth and maximum EPS production were clearly influenced by temperature and pH, the specific EPS production was only affected by stress conditions (T = 49 degrees C). CONCLUSIONS: Addition of whey protein hydrolysate to milk medium resulted in an increased growth and EPS production of S. thermophilus ST 111 under pH-controlled conditions. A modelling approach allowed studying the influence of temperature and pH on the kinetics of both growth and EPS production. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of an appropriate milk based medium and a combined model of temperature and pH can be of practical importance for the production of yoghurt or other fermented milks as well as for process optimization of the large-scale production of starter strains to be used for their EPS production. PMID- 15546418 TI - Characterization of Saccharomyces cerevisiae strains isolated from must of grape grown in experimental vineyard. AB - AIMS: Isolation and characterization of indigenous Saccharomyces cerevisiae strains from 12 grape varieties grown in an experimental vineyard of Apulia. METHODS AND RESULTS: Thirty to 40 colonies from each of the 12 fermentations were obtained at the end stage of spontaneous fermentation. By using morphological and physiological methods and by the PCR analysis of internal transcribed ITS1-5,8S ITS2, the isolates belonging to Saccharomyces genus were identified. These isolates were further characterized by amplification with S. cerevisiae species- and delta element-specific primers, thus allowing the identification of S. cerevisiae strains selected from each of the 12 fermentations. By means of RFLP analysis of mtDNA, each S. cerevisiae population isolated from a single fermentation appeared to constitute a genetically homogenous group. The comparison of the 12 cultivar-specific mtDNA RFLP patterns, allowed classifying the 12 S. cerevisiae populations into three genetically homogenous groups. The isolated strains fermented vigorously in synthetic and grape juice medium and showed high alcohol and sulphur dioxide (SO(2)) resistance and low hydrogen sulphite (H(2)S) production. CONCLUSIONS: The molecular analysis, in conjunction with the traditional morphological and physiological methods, was useful in discriminating at strain level the indigenous population of S. cerevisiae present in a vineyard of Apulia. The dominant S. cerevisiae strains identified in the 12 fermented musts showed potentially important oenological characteristics. SIGNIFICANCE AND IMPACT OF THE STUDY: The characterization of natural S. cerevisiae strains from several typical Italian grapes grown in a restricted experimental vineyard is an important step towards the preservation and exploitation of yeast biodiversity of Apulia, a relevant wine-producing region. The close relationship between the S. cerevisiae strains from different grapes grown in the same vineyard indicated that the occurrence of native strains is representative of the area rather than of the variety of grapes. PMID- 15546419 TI - Relationship between inactivation kinetics of a Listeria monocytogenes suspension by chlorine and its chlorine demand. AB - AIMS: Chlorine demand by Listeria monocytogenes cells and inactivation of L. monocytogenes by chlorine (0.6-1.0 mg l(-1)) at different temperatures (4, 20 and 30 degrees C) have been investigated in a batch reactor. METHODS AND RESULTS: Chlorine demand depended on the microbial concentration and was independent on the initial chlorine concentration and temperature. Chlorine decay was modelled by the addition of two first-order decay equations. Inactivation of L. monocytogenes by chlorine depended on the initial microbial concentration, initial chlorine concentration and temperature. A mathematical model based on a biphasic inactivation properly described survival curves of L. monocytogenes and a tertiary model was developed that satisfactorily predicted the inactivation of L. monocytogenes by different concentrations of initial chlorine at different temperatures. CONCLUSIONS: Both available chlorine decay and inactivation of L. monocytogenes by chlorine were biphasic and can be modelled by a two-term exponential model. SIGNIFICANCE AND IMPACT OF THE STUDY: The biphasic nature of survival curves of L. monocytogenes did not reflect the effect of a change of available chlorine concentration during the treatment. The microbial inactivation was caused by successive reactions that occur after the consumption of the chlorine by the bacterial cell components. PMID- 15546420 TI - Antifungal effects of herbal essential oils alone and in combination with ketoconazole against Trichophyton spp. AB - AIMS: To determine the effects of herbal essential oils on Trichophyton spp. growth and to evaluate the effects of Pelargonium graveolens oil and its main components citronellol and geraniol combined with ketoconazole against Trichophyton spp. METHODS AND RESULTS: Growth inhibition of six Trichophyton spp. by herbal essential oils was accessed and the combined effects of P. graveolens oil and its main components citronellol and geraniol were evaluated using a checkerboard microtitre assay against T. schoenleinii, T. erinacei and T. soudanense. The essential oil fraction of P. graveolens and its main components, geraniol and citronellol, exhibited strong synergism with ketoconazole against T. schoenleinii and T. soudanense, with fractional inhibitory concentration (FIC) indices in the range of 0.18-0.38. CONCLUSIONS: The antifungal effects of ketoconazole against Trichophyton spp. are enhanced significantly by administering it in combination with the essential oil fraction of P. graveolens or its main components, because of strong synergism, especially against T. soudanense and T. schoenleinii. SIGNIFICANCE AND IMPACT OF THE STUDY: The combination of ketoconazole and the essential oil fraction from P. graveolens or its main components for treatment of infections caused by Trichophyton species may reduce the minimum effective dose of ketoconazole, and thus minimize the side effects of ketoconazole. PMID- 15546421 TI - Isolation of a lactic acid bacterium and yeast consortium from a fermented material of Ulva spp. (Chlorophyta). AB - AIMS: Microbiota in a fermented culture of Ulva spp. was examined with the objective to characterize the type of fermentation and to obtain starter microbes for performing seaweed fermentation. METHOD AND RESULTS: Fermented Ulva spp. cultures which were obtained and transferred in a laboratory were examined for their microbiota. With phenotypic characterization and phylogenetic analysis based on rRNA gene nucleotide sequences, the predominant micro-organisms were identified as Lactobacillus brevis, Debaryomyces hanseni var. hansenii, and a Candida zeylanoides-related specimen, suggesting that the observed fermentation can be categorized to lactic acid and ethanol fermentation. Inoculating the individually cultured cell suspensions of the three kinds of micro-organisms with cellulase induced the fermentation in various kinds of seaweed. CONCLUSIONS: A microbial consortium composed of a lactic acid bacterium, L. brevis, and yeasts, D. hansenii and a C. zeylanoides-related specimen, were predominant in a fermented culture of Ulva spp. Lactic acid and ethanol fermentation could be induced in various kinds of seaweed by adding this microbial consortium along with cellulase. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of lactic acid and ethanol fermentation in seaweed, which is expected to provide a new material for food and dietary applications. PMID- 15546422 TI - Identification of early microbial colonizers in human dental biofilm. AB - AIMS: To elucidate the first colonizers within in vivo dental biofilm and to establish potential population shifts that occur during the early phases of biofilm formation. METHODS AND RESULTS: A 'checkerboard' DNA-DNA hybridization assay was employed to identify 40 different bacterial strains. Dental biofilm samples were collected from 15 healthy subjects, 0, 2, 4 and 6 h after tooth cleaning and the composition of these samples was compared with that of whole saliva collected from the same individuals. The bacterial distribution in biofilm samples was distinct from that in saliva, confirming the selectivity of the adhesion process. In the very early stages, the predominant tooth colonizers were found to be Actinomyces species. The relative proportion of streptococci, in particular Streptococcus mitis and S. oralis, increased at the expense of Actinomyces species between 2 and 6 h while the absolute level of Actinomyces remained unaltered. Periodontal pathogens such as Tannerella forsythensis(Bacteroides forsythus), Porphyromonas gingivalis and Treponema denticola as well as Actinobacillus actinomycetemcomitans were present in extremely low levels at all the examined time intervals in this healthy group of subjects. CONCLUSION: The data provide a detailed insight into the bacterial population shifts occurring within the first few hours of biofilm formation and show that the early colonizers of the tooth surface predominantly consist of beneficial micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: The early colonizers of dental plaque are of great importance in the succession stages of biofilm formation and its overall effect on the oral health of the host. PMID- 15546423 TI - Analysis of Acanthamoeba polyphaga surface carbohydrate exposure by FITC-lectin binding and fluorescence evaluation. AB - AIMS: Characterization of the representative protozoan Acanthamoeba polyphaga surface carbohydrate exposure by a novel combination of flow cytometry and ligand receptor analysis. METHODS AND RESULTS: Trophozoite and cyst morphological forms were exposed to a panel of FITC-lectins. Population fluorescence associated with FITC-lectin binding to acanthamoebal surface moieties was ascertained by flow cytometry. Increasing concentrations of representative FITC-lectins, saturation binding and determination of K(d) and relative B(max) values were employed to characterize carbohydrate residue exposure. FITC-lectins specific for N acetylglucosamine, N-acetylgalactosamine and mannose/glucose were readily bound by trophozoite and cyst surfaces. Minor incremental increases in FITC-lectin concentration resulted in significant differences in surface fluorescence intensity and supported the calculation of ligand-binding determinants, K(d) and relative B(max), which gave a trophozoite and cyst rank order of lectin affinity and surface receptor presence. CONCLUSIONS: Trophozoites and cysts expose similar surface carbohydrate residues, foremost amongst which is N-acetylglucosamine, in varying orientation and availability. SIGNIFICANCE AND IMPACT OF THE STUDY: The outlined versatile combination of flow cytometry and ligand-receptor analysis allowed the characterization of surface carbohydrate exposure by protozoan morphological forms and in turn will support a valid comparison of carbohydrate exposure by other single-cell protozoa and eucaryotic microbes analysed in the same manner. PMID- 15546424 TI - PIBWin -- software for probabilistic identification. PMID- 15546426 TI - The challenges of obesity management. PMID- 15546427 TI - Canadian dietitians' views and practices regarding obesity and weight management. AB - OBJECTIVE: To provide insight into Canadian dietitians' attitudes and practices regarding obesity and weight management. DESIGN: Cross-sectional mail survey of a stratified random sample of members of Canadian dietetic associations. SUBJECTS: A total of 514 dietitians (74% of those surveyed), 350 (69%) of whom actively counselled overweight/obese clients. MEASUREMENTS: Participants received a questionnaire to assess dietitians' attitudes regarding obesity and overweight, views regarding their role in weight management, counselling practices, and the criteria used to judge success. Demographic variables were collected. RESULTS: Most dietitians believed that obesity contributes to morbidity and mortality, and that small weight losses produced important health benefits. However, 80% agreed that health indicators other than weight loss should be the focus of obesity management, and 55% specifically recommended that clients not weigh themselves. Instead, weight management was promoted by recommending healthy eating and increased physical activity. Three-quarters agreed that they are the profession best trained to manage obesity but two-thirds believed their time would be better spent preventing rather than managing obesity. Dietitians most valued education received from on-the-job support and mentoring from other dietitians. Participants reported wanting to learn more about motivational and behavioural modification counselling techniques. CONCLUSIONS: Canadian dietitians follow a lifestyle approach to weight management. Studies are required to formally assess the effectiveness of various aspects of this approach. PMID- 15546428 TI - Attitudes and barriers to dietary advice aimed at reducing risk of type 2 diabetes in first-degree relatives of patients with type 2 diabetes. AB - OBJECTIVE: To evaluate the attitudes to and adoption of dietary advice in nondiabetic first-degree relatives of patients with type 2 diabetes and to examine barriers to adherence. DESIGN: One-year controlled intervention study, where treatment group (n=73) received lifestyle education. Attitudes towards dietary advice, change in dietary habits and importance of potential barriers to adherence were evaluated by questionnaires. Nondiabetic relatives (25-55 years; males and females) of individuals with type 2 diabetes were recruited. Education was based on current nutrition recommendations and aimed at improving dietary fat quality, increasing intake of fruit and vegetables, with additional advice to reduce dietary glycaemic index (GI). MAIN OUTCOME MEASURES: Attitudes and importance of barriers were classified by the intervened subjects into four categories ranging from 'No problem' to 'Yes, definitely a problem'. Dietary adherence was monitored by food frequency questionnaire at baseline and after 1 year. RESULTS: Participants were generally in favour of advice aimed at improving dietary fat quality. Attitudes towards advice to reduce GI varied widely. Food selection changed in accordance with predefined dietary goals. 'Forgetfulness', 'low availability in lunch restaurant' and 'lack of ideas for cooking' were barriers to adherence. CONCLUSIONS: Dietary advice aimed at reducing risk of type 2 diabetes was generally positively received and adopted in subjects with heredity for the disease. The most prevalent barriers reported are potentially modifiable. PMID- 15546429 TI - A comparison of food group variety between toddlers with and without cystic fibrosis. AB - OBJECTIVES: To compare the food group variety and nutritional adequacy of the diet between toddlers with cystic fibrosis (CF) and age-matched controls. Subjects A clinical sample of 22 toddlers with CF (mean age=21.3 +/- 7.2 months) matched to a community sample of 22 healthy peers. MAIN OUTCOME MEASURES: The variety index for toddlers (VIT) and the mean adequacy ratio (MAR). RESULTS: Fruit group scores were highest for children with CF (0.95 +/- 0.13; possible range 0.00-1.00), and dairy group scores were highest for controls (0.90 +/- 0.18). All children earned the lowest scores for vegetables (CF: 0.15 +/- 0.12; controls: 0.26 +/- 0.22). No significant differences were found when comparing VIT and MAR scores by sample (P >0.05). A moderate positive relationship was found between total VIT scores and MAR scores for all children (r=0.38, P <0.05). Toddlers with CF did not achieve the 120-150% of Recommended Dietary Allowance (RDA) for energy. CONCLUSIONS: Toddlers with CF are consuming diets that are varied and nutritionally adequate for a healthy child. To optimize nutritional status and growth, current recommendations for toddlers with CF to eat a well balanced diet and to exceed daily RDA for energy requirements remain key dietary issues. PMID- 15546430 TI - Influence of individually estimated portion size on the assessment of nutritional risk in colorectal cancer in Portugal. AB - We evaluated the influence of individually estimated portion sizes on the estimate of nutrient related risk of colorectal cancer, using data from a Portuguese hospital based case-control study on diet and colorectal cancer. A total of 100 patients with colorectal adenocarcinoma (aged 15-92 years) and 211 controls (aged 36-89 years) were included. Two data sets were created for nutrient analysis, the first one allowed estimates of food intake using data on portion size as collected with visual aids during the interview. The second estimate substituted respondents' estimate with a standard portion size, as used in the semi-quantitative (SQ) food frequency approach. The two analytic approaches yielded similar energy and nutrient intakes in cases and controls. The percent range of concordance is acceptable, in the same quartile varying from 44 to 82% (mean: 56%) and very good in the same or adjacent (+/-1) quartile (mean: 91%, range: 85-97%). The two estimates lead to a similar pattern of multivariate odd's ratio, however the SQ estimates resulted in more significant findings. We conclude that little extra information is gained by including individual portion size information when assessing diet-related risk of colorectal cancer. PMID- 15546431 TI - Home enteral feeding audit. AB - OBJECTIVE: To determine the main problems that families experience in the first month after their child is discharged from hospital on home enteral tube feeds (HETF) in terms of obtaining feed and equipment, managing tube feeding and accessing health professionals. METHODS: A multiple choice/short answer questionnaire was used to interview 81 parents/carers of children aged 0-16 years discharged from a supra regional children's hospital on HETF for the first time between December 2001 and August 2003. Questions addressed issues such as: contact with the home delivery company (HDC); delivery of feed and equipment; training; ability to access help; illnesses associated with tube feeding; and understanding of procedures post-discharge. RESULTS: The main problems identified were: delayed first deliveries (> or =7 days post-discharge; 47%); equipment missing from the first delivery (41%); difficulty obtaining a prescription from the general practitioner (GP) (17%); changing to different equipment post discharge with minimal training on the new equipment (26%). CONCLUSIONS: There were many problems with delays in delivery of equipment, incorrect equipment and changing of equipment when patients were first discharged on HETF. Significant improvements are necessary in organization of home enteral feeding systems when patients are first discharged. PMID- 15546432 TI - An elemental diet for bowel obstruction in pregnancy: a case study. AB - Bowel obstruction is a recognised complication in pregnancy, especially in those with adhesions from previous surgery. We report a case where the use of an elemental diet was successful in avoiding surgery in a pregnant patient with recurrent conservatively managed small bowel obstruction. PMID- 15546433 TI - Advice available on the Internet for people with coeliac disease: an evaluation of the quality of websites. AB - BACKGROUND: The Internet may be a useful resource for people with coeliac disease as a great deal of health-related information is published online. However, not all of it is accurate. It has been suggested that accurate information is most likely found on transparent sites and kitemarks are awarded on this basis. This paper examines whether the Internet is a useful resource for people with coeliac disease and whether transparency criteria can be used in identifying accurate sites. METHOD: An evaluation tool was developed using selected transparency criteria and clinical guidelines for accuracy. A total of 63 websites were evaluated. RESULTS: In the study, 66% of the websites scored less than 50% for accuracy. This was primarily because of incomplete information but 15.9% of sites contained inaccuracies. Over 50% of sites scored less than 50% for transparency. No correlation was found between sites that scored highly for accuracy and those that scored highly for transparency. CONCLUSION: There are useful information available for people with coeliac disease but transparency criteria alone cannot be used to identify accurate sites. PMID- 15546434 TI - Increased intake of fruit and vegetables and a low-fat diet, with and without low fat plant sterol-enriched spread consumption: effects on plasma lipoprotein and carotenoid metabolism. AB - BACKGROUND: Regular intake of plant sterol (phytosterol)-enriched foods enhances the cholesterol lowering effect of diets. One side effect associated with plant sterol consumption is a modest reduction in plasma carotenoid concentrations. This study investigated the effect of consuming a low-fat National Cholesterol Education Programme (NCEP) Step 1 diet, including a low-fat plant sterol ester (PSE)-enriched spread on cholesterol metabolism to determine if specific dietary advice to increase daily fruit and vegetable intake could prevent reduced plasma carotenoid concentrations. MATERIALS AND METHODS: In this randomised, crossover double-blind trial, 48 hypercholesterolaemic men received 21 g day(-1) of a low fat PSE-enriched spread or placebo for 3 weeks, interrupted by 3 weeks washout. Individuals also adhered to a NCEP Step 1 diet and repeated 3-day food diaries monitored adherence. Specific advice was provided to increase dietary fruit and vegetable intakes. Fasting blood samples were collected at pre- and post intervention for lipoprotein and carotenoid analysis. RESULTS: Plasma total and low-density lipoprotein (LDL) cholesterol concentrations were significantly (P <0.05) reduced, by 4.6 and 7.1%, respectively, after the PSE-enriched low-fat spread. Plasma apo B concentrations were significantly (P <0.0005) lower after the PSE spread. PSE consumption was also associated with significantly (P <0.05) lower total plasma beta-carotene concentrations, but this change was not significant after lipid standardisation. PSE consumption had no effect on retinol, alpha-carotene, gamma-tocopherol, alpha-tocopherol, lutein, zeaxanthin, beta-crypyoxanthin or lycopene concentrations. CONCLUSION: Dietary advice to increase daily fruit and vegetable consumption may be effective in preventing a reduction in plasma carotenoid concentrations previously associated with PSE consumption. Further, PSE incorporated in a low-fat spread and consumed as part of a NCEP Step 1 diet are effective in reducing total and LDL cholesterol. PMID- 15546450 TI - Dispelling the myths about ankylosing spondylitis. PMID- 15546451 TI - Clinical features in the emergency department can identify patients with suspected acute coronary syndromes who are safe for care in unmonitored hospital beds. AB - BACKGROUND: Standard practice for patients requiring hospital admission with suspected acute coronary syndromes (ACS) is admission to a monitored cardiology bed. The Western Hospital Chest Pain Protocol was developed to identify a subset of these patients who could be safely managed in an unmonitored bed. AIM: The objective of this prospective study of chest pain patients classified as 'high' or 'intermediate' risk by the Agency for Health Care Policy and Research/National Health and Medical Research Council guidelines was to further evaluate the safety of this protocol. METHODS: This study was a prospective, observational, cohort study investigating the outcomes of patients admitted to hospital with suspected ACS. The primary outcome of interest was death or life-threatening arrhythmia within 24 h of hospital admission. RESULTS: If the Western Hospital Chest Pain Protocol had been strictly applied, there would have been one death in the group assigned to unmonitored beds (1/750; 0.13%, 95% confidence interval 0.01-0.85%) and no other life-threatening arrhythmias. CONCLUSION: There is a subgroup of patients with suspected ACS who require hospital admission who can, based on clinical and biochemical features in the emergency department, be safely assigned to unmonitored beds. PMID- 15546452 TI - Trust and confidence: towards mutual acceptance of ethics committee approval of multicentre studies. AB - AIMS: To compare issues raised by Human Research Ethics Committees (HREC) during the ethics review process and to determine the length of time taken to gain HREC approval for multicentre research studies. METHODS: Review and analysis of HREC documentation and correspondence for all multicentre research studies were conducted through three HREC under the auspices of Cancer Trials Australia, Melbourne, Victoria, Australia, between November 1997 and March 2001 to determine the variance of documentation, correspondence and recommendations across the three HREC and the time taken for study approval. RESULTS: Thirty-one projects were submitted to any two of the HREC (16 studies) or all three HREC (15 studies). The median time for study approval at an individual HREC was 75 days, but it was 111 days for approval at all participating sites. There were 554 clarifications or comments made by the reviewing HREC, the majority of which had no significant bearing on the ethical or scientific calibre of the study. There was only one study in which a significant protocol change was requested by a HREC. CONCLUSIONS: Multicentre study approvals are delayed when submitted to multiple HREC. The three HREC raised similar issues without substantive differences in their recommendations. A process for the mutual acceptance of HREC recommendations could facilitate multicentre research. PMID- 15546453 TI - Unobvious wounds: the suffering of hospice patients. AB - BACKGROUND: The suffering of palliative care patients is ordinarily thought of in terms of symptoms, and these are identified by medical terms (pain, nausea, depression). The problematic issue of relief of patients' pain meaning the same thing as the 'relief of the suffering and distress of terminally ill patients' has been raised. AIM: The aim of this study was to estimate the frequency with which medically defined suffering matched the reported suffering of our patients. METHODS: One hundred patients admitted to a hospice were asked 'In what way are you suffering?' The patients' diagnoses, their pain scores and the reasons for admission as defined by the treating clinicians were recorded. The mean age of the patients was 68 years (range 28-93 years), 92 had advanced malignant disease and 51 were women. RESULTS: Twenty-four patients were unable to state the reason for hospice admission, but none had any uncertainty in identifying the nature of their own suffering. There was a weak correlation between the patient's view of their suffering and the reason for admission. The identification of pain as the cause of suffering was weakly correlated with pain scores. Some patients with pain scores of 8-10/10 did not mention pain as a cause of suffering, and others with scores of 0/10 did identify pain as the cause of suffering. CONCLUSION: Asking hospice patients about suffering in a simple open-ended way can expose a different dimension of distress, and the views of the 100 patients of this study support the statement that relief of pain and relief of suffering are not the same. PMID- 15546454 TI - A chronic disease management programme can reduce days in hospital for patients with chronic obstructive pulmonary disease. AB - BACKGROUND: A steady increase in chronic obstructive pulmonary disease (COPD) admissions was addressed by enhancing primary care to provide intensive chronic disease management. AIM: To compare the effect of a disease management programme, including a COPD management guideline, a patient-specific care plan and collaboration between patients, general practitioners, practice nurses, hospital physicians and nurse specialists with conventional care, on hospital admissions and quality of life. METHODS: One hundred and thirty-five patients with a clinical diagnosis of moderate to severe COPD were identified from hospital admission data and general practice records. General practices were randomized to either conventional care (CON), or the intervention (INT). Pre- and post-study assessment included spirometry, Shuttle Walk Test, Short Form-36, and the Chronic Respiratory Questionnaire (CRQ). Admission data were compared for 12 months prior to and during the trial. RESULTS: For respiratory conditions, mean hospital bed days per patient per year for the INT group were reduced from 2.8 to 1.1, whereas those for the CON group increased from 3.5 to 4.0 (group difference, P = 0.030) The INT group also showed an improvement for two dimensions of the CRQ, fatigue (P = 0.010) and mastery (P = 0.007). CONCLUSIONS: A chronic disease management programme for COPD patients that incorporated a variety of interventions, including pulmonary rehabilitation and implemented by primary care, reduced admissions and hospital bed days. Key elements were patient participation and information sharing among healthcare providers. PMID- 15546455 TI - Reliability of X-rays and bone scans for the assessment of changes in skeletal metastases from breast cancer. AB - AIMS: To examine the level of agreement among observers regarding changes between serial images of bone metastases. METHODS: Thirty-five pairs of bone X-rays and 30 pairs of bone scans were selected from the files of patients with breast cancer involving the skeleton. All images in a pair were of the same site and had been taken at least 12 weeks apart. Thirteen radiologists and 14 nuclear medicine physicians examined the X-ray and bone scan pairs, respectively. Each assessed whether the changes between sequential films represented improvement, stability or worsening. Inter-observer agreement was analysed using the kappa statistic (kappa). RESULTS: There was only fair overall agreement among radiologists regarding changes between X-rays (kappa = 0.23), but there was substantial agreement among nuclear medicine physicians for bone scan assessments (kappa = 0.62). Neither the experience of the observers nor the time between images had a significant effect on agreement. For X-rays, agreement was poorer if the response category was 'improvement' and if the type of bone lesion was mixed lytic/sclerotic. CONCLUSIONS: Evaluation of serial X-rays is unreliable for determining the response of bone metastases. Scintigraphic evaluation has a higher internal validity for the determination of response, but it should not be used in isolation from other clinical data. PMID- 15546456 TI - Gene therapy: applications and progress towards the clinic. AB - Gene therapy was originally conceived as an approach to the treatment of genetic disease, to repair or replace a faulty gene. Subsequently, gene therapy clinical trials have been undertaken for a wide range of conditions, particularly cancer and AIDS. Overall, the results from gene therapy have been disappointing. The reasons include the following: (i) low gene transfer efficiencies and (ii) shortcomings in the identification and manipulation of appropriate target cells, including progenitor cell populations required for the maintenance of long-term effects. Today, the immense potential of gene therapy remains, but more basic research is required to improve technical aspects of this form of cellular therapy. PMID- 15546457 TI - Fibrosing alveolitis in systemic sclerosis: the need for early screening and treatment. AB - Abnormalities in lung function occur in 70% of patients with systemic sclerosis (SSc). Fibrosing alveolitis in SSc (FASSc) is more commonly seen in the diffuse cutaneous form of SSc, particularly in the presence of antitopoisomerase antibodies (Scl70), and with the decreasing incidence of scleroderma renal crisis it is now the major cause of mortality in this patient population. Screening of patients recently diagnosed with SSc by pulmonary function tests and the performance of high resolution computed tomography when physiological abnormalities are identified has resulted in the identification of significant numbers of patients with early, asymptomatic FASSc. Whether these patients should be further investigated with a surgical lung biopsy or receive immunosuppression is unclear, because it cannot yet be reliably predicted who will develop progressive disease and the evidence to support the efficacy of treatment is not strong. The objective of the present article was to review the evidence to support the use of immunosuppressive therapy in FASSc and, based on these data, to propose an algorithm for the investigation and management of this difficult clinical problem. PMID- 15546458 TI - Audit of community-based anticoagulant monitoring in patients with thromboembolic disease: is frequent testing necessary? AB - Oral anticoagulant monitoring is managed by general practitioners in Auckland. An audit of this service in 452 patients demonstrated that anticoagulant control was in line with recommended international guidelines, with 58.3% of international normalized ratio (INR) measurements in the therapeutic range. However, the frequency of testing was high, with the majority of patients (68%), including those on long-term treatment, having INR measurements at weekly intervals. We question the need for such frequent INR testing. PMID- 15546459 TI - Five cases of non-typhoidal Salmonella endovascular infection. AB - The incidence of human non-typhoidal Salmonella (NTS) infection has increased in many countries. Endovascular infection is one of the most serious forms of extraintestinal infection. Five patients with NTS endovascular infection treated at Middlemore Hospital, Auckland, New Zealand, are presented here. All persons with NTS bacteraemia who are older than 50 years and have a risk of atherosclerosis should be evaluated for possible endovascular infection. PMID- 15546460 TI - 'Is she experiencing any pain?': disability and the physician-patient relationship. AB - In this essay, I reflect on my experience as a multiple sclerosis patient in order to identify some of the unique challenges that chronic disability poses in the physician-patient relationship. I suggest that it is important to broaden the goals of the clinical encounter to incorporate personal (as opposed to simply bodily) well-being, to be aware of the manner in which chronic disability affects decisions regarding treatment, to recognize the import of physical and attitudinal barriers and to acknowledge that patients with chronic disabilities have an 'expert' knowledge of bodily experience. I also suggest that chronic disability provides an exceptional opportunity with respect to the relationship between doctors and patients. PMID- 15546461 TI - Impact of selective cyclooxygenase-2 inhibitors on anti-ulcer medication and non steroidal anti-inflammatory drug use in patients with rheumatic disease. PMID- 15546463 TI - Diagnosis of a radiographic right chest opacity. PMID- 15546464 TI - Splenic rupture after colonoscopy. PMID- 15546465 TI - Severe alpha-1 antitrypsin deficiency diagnosed in an 86-year-old man. PMID- 15546466 TI - Unusual presentations of giant cell arteritis (temporal arteritis). PMID- 15546467 TI - Altered calcium homeostasis during pregnancy may affect biochemical differentiation of hypercalcaemia. PMID- 15546473 TI - Current in vivo assays for cutaneous toxicity: local and systemic toxicity testing. AB - This review provides an update on current (June 2004) in vivo assays for evaluations of local and systemic cutaneous toxicity. As the science of toxicology and safety assessment grows and evolves, an increasing number of in vitro assays and screening procedures designed to minimize the use of animals in cutaneous toxicity evaluations have been validated or are currently in development. However, in some cases, it remains necessary to evaluate the effects of new drugs or chemicals in an animal model in order to protect human health. Current regulatory requirements and study designs for local and systemic in vivo cutaneous toxicity evaluations, as well as discussions of procedures unique to cutaneous toxicity evaluation are presented in this review. PMID- 15546474 TI - Mesenchymal stem cells: cell biology and potential use in therapy. AB - Mesenchymal stem cells are clonogenic, non-haematopoietic stem cells present in the bone marrow and are able to differentiate into multiple mesoderm-type cell lineages e.g. osteoblasts, chondrocytes, endothelial-cells and also non-mesoderm type lineages e.g. neuronal-like cells. Several methods are currently available for isolation of the mesenchymal stem cells based on their physical and immunological characteristics. Because of the ease of their isolation and their extensive differentiation potential, mesenchymal stem cells are among the first stem cell types to be introduced in the clinic. Recent studies have demonstrated that the life span of mesenchymal stem cells in vitro can be extended by increasing the levels of telomerase expression in the cells and thus allowing culture of large number of cells needed for therapy. In addition, it has been shown that it is possible to culture the cells in xeno-free environment without affecting their growth or differentiation potential. Finally, the mesenchymal stem cells seems to be hypoimmunogenic and thus allogenic mesenchymal stem cells transplantation is possible. It is envisaged that mesenchymal stem cells can be used in systemic transplantation for generalized diseases, local implantation for local tissue defects, as a vehicle for genes in gene therapy protocols or to generate transplantable tissues and organs in tissue engineering protocols. The results of these initial trials are very encouraging and several clinical trials are under way to study the efficacy and long-term safety of therapeutics based on mesenchymal stem cells. PMID- 15546475 TI - Inhibition of the protease activity of the light chain of type A botulinum neurotoxin by aqueous extract from stinging nettle (Urtica dioica) leaf. AB - We investigated the inhibitory effect of stinging nettle leaf extract on the protease activity of botulinum neurotoxin type A and B light chains. The nettle leaf infusion was fractionated and HPLC-based enzymatic assays were performed to determine the capacity of each fraction to inhibit the protease activity of botulinum neurotoxin type A and B light chains. Assay results demonstrated that a water-soluble fraction obtained from the nettle leaf infusion inhibited type A, but did not inhibit type B light chain protease activity. The inhibition mode of water soluble fraction against protease activity of type A light chain was analyzed and found to be a non-competitive. PMID- 15546476 TI - Effect of Hibiscus rosa sinensis extract on hyperproliferation and oxidative damage caused by benzoyl peroxide and ultraviolet radiations in mouse skin. AB - The present study was conducted to investigate the ameliorative potential of Hibiscus rosa sinensis extract in mice skin. Combination of a single topical application of benzoyl peroxide (20 mg/0.2 ml/animal) followed by ultraviolet radiations (0.420 J/m2/s) was used to induce hyperproliferation and oxidative stress. Single benzoyl peroxide application prior to ultraviolet B radiations exposure caused significant depletion in the detoxification and antioxidant enzymes, while malondialdehyde formation, hydrogen peroxide content, ornithine decarboxylase activity and DNA synthesis were raised significantly. However, pretreatment of H. rosa sinensis extract (3.5 mg and 7 mg/ kg b.wt.) partly restored the levels of cellular protective enzymes (P<0.05). Besides, malondialdehyde formation and hydrogen peroxide content (P<0.05) were statistically significantly reduced at both doses. The ornithine decarboxylase activity and thymidine incorporation in DNA were also reduced dose dependently (P<0.05) by the plant extract. Therefore, we propose that H. rosa sinensis extract exerts a protective effect against the tumour promotion stage of cancer development. PMID- 15546477 TI - Inhibitory action of ethanol on cyclooxygenase-2 expression through suppression of the extracellular signal-related kinase-mediated pathway in rat alveolar macrophages. AB - The purpose of the present study was to elucidate the effects of ethanol on expression of cyclooxygenase in alveolar macrophages. Rat alveolar macrophages were collected by bronchoalveolar lavage and stimulated by lipopolysaccharide. Lipopolysaccharide-induced cyclooxygenase-2 expression and prostaglandin E2 production were inhibited by ethanol (100-200 mM) in a concentration-dependent manner. Ethanol at 100-200 mM concentration-dependently inhibited cyclooxygenase 2 mRNA expression. Lipopolysaccharide-stimulated phosphorylation of both extracellular signal-related kinase and p38 mitogen-activated protein kinase was also significantly inhibited by ethanol (50-200 mM). Cyclooxygenase-2 expression was significantly inhibited by U0126 but was not affected by SB203580. In the presence of SB203580, lipopolysaccharide-induced cyclooxygenase-2 expression was also inhibited by ethanol (50-200 mM). On the other hand, cyclooxygenase-1 was expressed constitutively in alveolar macrophages and cyclooxygenase-1 expression was affected neither by lipopolysaccharide nor ethanol. Lipopolysaccharide induced expression of inducible nitric oxide synthase in alveolar macrophages was not affected by ethanol at 50-200 mM. These results suggest that lipopolysaccharide-induced expression of cyclooxygenase-2 is mediated by extracellular signal-related kinase but not by p38 kinase and that ethanol selectively attenuates cyclooxygenase-2 expression mainly by inhibiting activation of extracellular signal-related kinase. PMID- 15546478 TI - Effects of anticancer chemotherapeutic drugs on the acetylcholine receptor operated potassium current in guinea pig atrial myocytes. AB - The effects of 7 anticancer chemotherapeutic drugs on the muscarinic acetylcholine receptor-operated potassium current (I(K.ACh)) in guinea pig atrial myocytes were investigated using the whole cell patch clamp technique. Doxorubicin, pirarubicin, and mitoxantrone inhibited the carbachol-induced I(K.ACh) in a concentration-dependent manner in atrial cells at a holding potential of -40 mV. IC50 values of doxorubicin, pirarubicin, and mitoxantrone for the carbachol-induced I(K.ACh) were 7.7 microM, 3.7 microM, and 9.1 microM, respectively. Pirarubicin inhibited the adenosine-induced and the GTPgammaS induced I(K.ACh) in a concentration-dependent manner (IC50=6.0 and 5.1 microM, respectively). Doxorubicin and mitoxantrone up to 100 microM did not have an influence on the adenosine-induced I(K.ACh). Doxorubicin did not affect the GTPgammaS-induced I(K.ACh). Mitoxantrone 100 microM inhibited the current only by 25%. For concentrations up to 100 microM, anticancer drugs that have chemical structures entirely different from that of doxorubicin, i.e., 5-fluorouracil, 6 mercaptopurine, cyclophosphamide, and actinomycin D, did not have an influence on the carbachol-induced I(K.ACh). Doxorubicin and chemically related compounds possess anticholinergic effects mediated via an inhibitory action on I(K.ACh) by different underlying molecular mechanisms. Doxorubicin and mitoxantrone may inhibit I(K.ACh) by the blockade of muscarinic receptors, whereas pirarubicin may inhibit the current not only via blocking the muscarinic receptors but also by depressing the functions of the K+ channel itself and/or GTP-binding proteins. PMID- 15546479 TI - (+) MK-801 and phencyclidine induced neurotoxicity do not cause enduring behaviours resembling the positive and negative symptoms of schizophrenia in the rat. AB - Studies in rats and primates have demonstrated that repeated phencyclidine treatment can produce enduring cognitive deficits that may resemble the cognitive deficits seen in schizophrenia, suggesting that neurodegeneration resulting from NMDA-receptor dysfunction may be a valid model of schizophrenia. The purpose of the present experiments was to expand these findings and to determine if medium and high doses of the NMDA-antagonists phencyclidine and (+)MK-801 could produce permanent behavioural changes in animal tests with face validity for some aspects of the positive and negative symptoms of schizophrenia. Rats were treated with dose regimens of (+)MK-801 and phencyclidine known to produce mild and severe irreversible levels of neurotoxicity, and were tested 7 or 10 days after the last drug administration in the social interaction test and in standard activity cages. The rats did not show any enduring behavioural changes as a result of the treatment. The present study could therefore not provide additional evidence for the face validity of this model of schizophrenia. PMID- 15546480 TI - Standard of care may not protect against acetaminophen-induced nephrotoxicity. PMID- 15546481 TI - Testicular seminoma after the complete remission of extragonadal yolk sac tumor : a case report. AB - BACKGROUND: Between 2% and 5% of malignant germ-cell tumors in men arise at extragonadal sites. Of extragonadal germ cell tumors, testicular carcinoma in situ (CIS) are present in 31-42% of cases, and CIS are reported to have low sensitivity to chemotherapy in spite of the various morphology and to have a high likelihood of developing into testicular tumors. A testicular biopsy may thus be highly advisable when evaluating an extragonadal germ cell tumor. CASE PRESENTATION: A 36-year-old man was diagnosed as having an extragonadal non seminomatous germ cell tumor, that was treated by cisplatin-based chemotherapy, leading to a complete remission. In the meantime, testicular tumors were not detected by means of ultrasonography. About 4 years later, a right testicular tumor was found, and orchiectomy was carried out. Microscopically, the tumor was composed of seminoma. CONCLUSIONS: We herein report a case of metachronous occurrence of an extragonadal and gonadal germ cell tumor. In the evaluation of an extragonadal germ cell tumor, a histological examination should be included since ultrasonography is not sufficient to detect CIS or minute lesions of the testis. PMID- 15546482 TI - The association of patient trust and self-care among patients with diabetes mellitus. AB - BACKGROUND: Diabetes requires significant alterations to lifestyle and completion of self management tasks to obtain good control of disease. The objective of this study was to determine if patient trust is associated with reduced difficulty and hassles in altering lifestyle and completing self care tasks. METHODS: A cross sectional telephone survey and medical record review was performed to measure patient trust and difficulty in completing diabetes tasks among 320 medically underserved patients attending diabetes programs in rural North Carolina, USA. Diabetes tasks were measured three ways: perceived hassles of diabetic care activities, difficulty in completing diabetes-related care activities, and a global assessment of overall ability to complete diabetes care activities. The association of patient trust with self-management was examined after controlling for patient demographics, physical functioning, mental health and co-morbidities. RESULTS: Level of patient trust was high (median 22, possible max 25). Higher trust levels were associated with lower levels of hassles (p = 0.006) and lower difficulty in completing care activities (p = 0.001). Patients with higher trust had better global assessments of overall ability to complete diabetes care activities (p < 0.0001). CONCLUSION: Higher patient trust in physicians is associated with reduced difficulty in completing disease specific tasks by patients. Further studies are needed to determine the causal relationship of this association, the effect of trust on other outcomes, and the potential modifiability of trust. PMID- 15546483 TI - A national survey of the prevalence of schistosomiasis and soil transmitted helminths in Malawi. AB - BACKGROUND: Past estimates have put the prevalence of schistosomiasis between 40% and 50% in the Malawi population overall based on studies undertaken ten years or more ago. More recent surveys in known high risk areas find similar levels. However control measures, changing ecology and migration may have led to changes in the prevalence of schistosomiasis in different parts of Malawi. A national schistosomiasis and soil-transmitted helminth (STH) survey was undertaken to measure the distribution, prevalence and intensity of infection in November 2002. METHODS: A school was selected randomly from a random sample of 30 Traditional Authorities stratified by six distinct ecological zones, and 1,664 year 3 pupils (9-10 year olds) were questioned about recent illnesses and "red urine". Samples of urine and faeces were examined for the presence of eggs using the standard Kato-Katz technique for soil-transmitted helminths and intestinal schistosomiasis and urine samples using the filtration technique for Schistosoma haematobium. RESULTS: The prevalence of Schistosoma mansoni is 0.4% (95% CI 0-1.3%), S. haematobium 6.9% (95% CI 1.9 - 11.9%), hookworm 1.3% (95% CI 0.4-2.3%), Ascariasis 0.5% (95% CI 0.1-1.0%) and trichuriasis 0% in year 3 pupils (modal age 10 years of age). Intensity of infection is low for all infections except for 2.5% who have high intensity S. haematobium infection. The "red urine" question is 67% sensitive and 80% specific for positive S. haematobium microscopy. CONCLUSIONS: The reduction in prevalences may be real as a result of recent control measures, or false if historical results were based on surveys of high risk populations. Another explanation is that this survey used an unrepresentative sample of schools. Detailed analysis suggests this is unlikely. Recommendations include the use of a 30% positive threshold for the "red urine" screening question to be used in schoolchildren in high prevalence areas. This survey, based on a national probability sample excluding the northern region lakeside area, finds much lower overall prevalence and intensity of schistosomiasis and STHs than previous estimates based on selected surveys. Disease control featuring chemotherapy may be having a profound effect. The localised nature of the distribution of the infections means that control programmes may work best if undertaken at district level or below. "Red urine" questionnaire surveys may help identify hot spots. PMID- 15546484 TI - Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy - spatial and time dependent distribution. AB - BACKGROUND: Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. METHODS: Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP) and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. RESULTS: Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. CONCLUSIONS: Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous distribution of gene expression in the syngeneic P22 rat tumor model. PMID- 15546485 TI - Laboratory diagnosis and susceptibility profile of Helicobacter pylori infection in the Philippines. AB - BACKGROUND: Helicobacter pylori diagnosis and susceptibility profile directs the applicability of recommended treatment regimens in our setting. To our knowledge, there is no published data on the culture and local susceptibility pattern of Helicobacter pylori in the Philippines. METHODS: 52 dyspeptic adult patients undergoing endoscopy from the Outpatient Gastroenterology clinic of the University of the Philippines-Philippine General Hospital underwent multiple gastric biopsy and specimens were submitted for gram stain, culture, antimicrobial sensitivity testing, rapid urease test and histology. Antimicrobial susceptibility testing was done by Epsilometer testing (Etest) method against metronidazole, clarithromycin, amoxicillin, and tetracycline. RESULTS: Sixty percent (60%) of the study population was positive for H. pylori infection (mean age of 44 years +/- 13), 70% were males. H. pylori culture showed a sensitivity of 45% (95% CI [29.5-62.1]), specificity of 98% (95%CI [81.5-100%]), positive likelihood ratio of 19.93 (95% CI [1.254-317.04]) and a negative likelihood ratio of 0.56 (95% CI [0.406-0.772]). All H. pylori strains isolated were sensitive to metronidazole, clarithromycin, amoxicillin and tetracycline. CONCLUSION: Knowledge of the antibiotic susceptibility patterns in our setting allows us to be more cautious in the choice of first-line agents. Information on antibiotic susceptibility profile plays an important role in empiric antibiotic treatment and management of refractive cases. PMID- 15546486 TI - The rehydration transcriptome of the desiccation-tolerant bryophyte Tortula ruralis: transcript classification and analysis. AB - BACKGROUND: The cellular response of plants to water-deficits has both economic and evolutionary importance directly affecting plant productivity in agriculture and plant survival in the natural environment. Genes induced by water-deficit stress have been successfully enumerated in plants that are relatively sensitive to cellular dehydration, however we have little knowledge as to the adaptive role of these genes in establishing tolerance to water loss at the cellular level. Our approach to address this problem has been to investigate the genetic responses of plants that are capable of tolerating extremes of dehydration, in particular the desiccation-tolerant bryophyte, Tortula ruralis. To establish a sound basis for characterizing the Tortula genome in regards to desiccation tolerance, we analyzed 10,368 expressed sequence tags (ESTs) from rehydrated rapid-dried Tortula gametophytes, a stage previously determined to exhibit the maximum stress induced change in gene expression. RESULTS: The 10, 368 ESTs formed 5,563 EST clusters (contig groups representing individual genes) of which 3,321 (59.7%) exhibited similarity to genes present in the public databases and 2,242 were categorized as unknowns based on protein homology scores. The 3,321 clusters were classified by function using the Gene Ontology (GO) hierarchy and the KEGG database. The results indicate that the transcriptome contains a diverse population of transcripts that reflects, as expected, a period of metabolic upheaval in the gametophyte cells. Much of the emphasis within the transcriptome is centered on the protein synthetic machinery, ion and metabolite transport, and membrane biosynthesis and repair. Rehydrating gametophytes also have an abundance of transcripts that code for enzymes involved in oxidative stress metabolism and phosphorylating activities. The functional classifications reflect a remarkable consistency with what we have previously established with regards to the metabolic activities that are important in the recovery of the gametophytes from desiccation. A comparison of the GO distribution of Tortula clusters with an identical analysis of 9,981 clusters from the desiccation sensitive bryophyte species Physcomitrella patens, revealed, and accentuated, the differences between stressed and unstressed transcriptomes. Cross species sequence comparisons indicated that on the whole the Tortula clusters were more closely related to those from Physcomitrella than Arabidopsis (complete genome BLASTx comparison) although because of the differences in the databases there were more high scoring matches to the Arabidopsis sequences. The most abundant transcripts contained within the Tortula ESTs encode Late Embryogenesis Abundant (LEA) proteins that are normally associated with drying plant tissues. This suggests that LEAs may also play a role in recovery from desiccation when water is reintroduced into a dried tissue. CONCLUSION: The establishment of a rehydration EST collection for Tortula ruralis, an important plant model for plant stress responses and vegetative desiccation tolerance, is an important step in understanding the genome level response to cellular dehydration. The type of transcript analysis performed here has laid the foundation for more detailed functional and genome level analyses of the genes involved in desiccation tolerance in plants. PMID- 15546487 TI - Zygapophysial joint blocks in chronic low back pain: a test of Revel's model as a screening test. AB - BACKGROUND: Only controlled blocks are capable of confirming the zygapophysial joints (ZJ) as the pain generator in LBP patients. However, previous workers have found that a cluster of clinical signs ("Revel's criteria"), may be valuable in predicting the results of an initial screening ZJ block. It was suggested that these clinical findings are unsuitable for diagnosis, but may be of value in selecting patients for diagnostic blocks of the lumbar ZJ's. To constitute evidence in favour of a clinical management strategy, these results need confirmation. This study evaluates the utility of 'Revel's criteria' as a screening tool for selection of chronic low back pain patients for controlled ZJ diagnostic blocks. METHODS: This study utilized a prospective blinded concurrent reference standard related validity design. Consecutive chronic LBP patients completed pain drawings, psychosocial distress and disability questionnaires, received a clinical examination and lumbar zygapophysial blocks. Two reference standards were evaluated simultaneously: 1. 75% reduction of pain on a visual analogue scale (replication of previous work), and 2. abolition of the dominant or primary pain. Using "Revel's criteria" as predictors, logistic regression analyses were used to test the model. Estimates of sensitivity, specificity, predictive values and likelihood ratios for selected variables were calculated for the two proposed clinical strategies. RESULTS: Earlier results were not replicated. Sensitivity of "Revel's criteria" was low sensitivity (<17%), and specificity high (approximately 90%). Absence of pain with cough or sneeze just reached significance (p = 0.05) within one model. CONCLUSIONS: "Revel's criteria" are unsuitable as a clinical screening test to select chronic LBP patients for initial ZJ blocks. However, the criteria may have use in identifying a small subset (11%) of patients likely to respond to the initial block (specificity 93%). PMID- 15546488 TI - Treatment decision-making and the form of risk communication: results of a factorial survey. AB - BACKGROUND: Prospective users of preventive therapies often must evaluate complex information about therapeutic risks and benefits. The purpose of this study was to evaluate the effect of relative and absolute risk information on patient decision-making in scenarios typical of health information for patients. METHODS: Factorial experiments within a telephone survey of the Michigan adult, non institutionalized, English-speaking population. Average interview lasted 23 minutes. Subjects and sample design: 952 randomly selected adults within a random digit dial sample of Michigan households. Completion rate was 54.3%. RESULTS: When presented hypothetical information regarding additional risks of breast cancer from a medication to prevent a bone disease, respondents reduced their willingness to recommend a female friend take the medication compared to the baseline rate (66.8% = yes). The decrease was significantly greater with relative risk information. Additional benefit information regarding preventing heart disease from the medication increased willingness to recommend the medication to a female friend relative to the baseline scenario, but did not differ between absolute and relative risk formats. When information about both increased risk of breast cancer and reduced risk of heart disease were provided, typical respondents appeared to make rational decisions consistent with Expected Utility Theory, but the information presentation format affected choices. Those 11% - 33% making decisions contrary to the medical indications were more likely to be Hispanic, older, more educated, smokers, and to have children in the home. CONCLUSIONS: In scenarios typical of health risk information, relative risk information led respondents to make non-normative decisions that were "corrected" when the frame used absolute risk information. This population sample made generally rational decisions when presented with absolute risk information, even in the context of a telephone interview requiring remembering rates given. The lack of effect of gender and race suggests that a standard strategy of presenting absolute risk information may improve patient decision-making. PMID- 15546489 TI - beta-Amyloid promotes accumulation of lipid peroxides by inhibiting CD36-mediated clearance of oxidized lipoproteins. AB - BACKGROUND: Recent studies suggest that hypercholesterolemia, an established risk factor for atherosclerosis, is also a risk factor for Alzheimer's disease. The myeloid scavenger receptor CD36 binds oxidized lipoproteins that accumulate with hypercholesterolemia and mediates their clearance from the circulation and peripheral tissues. Recently, we demonstrated that CD36 also binds fibrillar beta amyloid and initiates a signaling cascade that regulates microglial recruitment and activation. As increased lipoprotein oxidation and accumulation of lipid peroxidation products have been reported in Alzheimer's disease, we investigated whether beta-amyloid altered oxidized lipoprotein clearance via CD36. METHODS: The availability of mice genetically deficient in class A (SRAI & II) and class B (CD36) scavenger receptors has facilitated studies to discriminate their individual actions. Using primary microglia and macrophages, we assessed the impact of Abeta on: (a) cholesterol ester accumulation by GC-MS and neutral lipid staining, (b) binding, uptake and degradation of 125I-labeled oxidized lipoproteins via CD36, SR-A and CD36/SR-A-independent pathways, (c) expression of SR-A and CD36. In addition, using mice with targeted deletions in essential kinases in the CD36-signaling cascade, we investigated whether Abeta-CD36 signaling altered metabolism of oxidized lipoproteins. RESULTS: In primary microglia and macrophages, Abeta inhibited binding, uptake and degradation of oxidized low density lipoprotein (oxLDL) in a dose-dependent manner. While untreated cells accumulated abundant cholesterol ester in the presence of oxLDL, cells treated with Abeta were devoid of cholesterol ester. Pretreatment of cells with Abeta did not affect subsequent degradation of oxidized lipoproteins, indicating that lysosomal accumulation of Abeta did not disrupt this degradation pathway. Using mice with targeted deletions of the scavenger receptors, we demonstrated that Abeta inhibited oxidized lipoprotein binding and its subsequent degradation via CD36, but not SRA, and this was independent of Abeta-CD36 signaling. Furthermore, Abeta treatment decreased CD36, but not SRA, mRNA and protein, thereby reducing cell surface expression of this oxLDL receptor. CONCLUSIONS: Together, these data demonstrate that in the presence of beta amyloid, CD36-mediated clearance of oxidized lipoproteins is abrogated, which would promote the extracellular accumulation of these pro-inflammatory lipids and perpetuate lipid peroxidation. PMID- 15546490 TI - Size control in growing yeast and mammalian cells. AB - BACKGROUND: In a recent publication it was claimed that cultured mammalian cells, in contrast to yeasts, maintain a constant size distribution in the population without a size checkpoint. This inference may be challengeable. RESULTS: (1) It is argued that "weak" size control implies the existence of a checkpoint, and unfortunately the technique used by Conlon and Raff might obscure such a weak mechanism. (2) Previous investigations of size control in yeasts have shown that individual cell data, rather than means and variances of cell populations, are prerequisites for reliable interpretation. (3) No experimental data so far obtained suggest that in any cell culture a linear growth pattern in cell mass can maintain size homeostasis on its own without size control. (4) Studies on fission yeast mutants indicate that the molecular mechanisms of size control vary with genetic background, implying that no single mechanism is likely to apply to any cell type, including cultured mammalian cells, under all conditions. CONCLUSION: The claim that cultured mammalian cells maintain size homeostasis without a checkpoint needs to be re-evaluated by measurements on individual cells. PMID- 15546491 TI - Application of methods of identifying receptor binding models and analysis of parameters. AB - BACKGROUND: Possible methods for distinguishing receptor binding models and analysing their parameters are considered. RESULTS AND DISCUSSION: The conjugate gradients method is shown to be optimal for solving problems of the kind considered. Convergence with experimental data is rapidly achieved with the appropriate model but not with alternative models. CONCLUSION: Lack of convergence using the conjugate gradients method can be taken to indicate inconsistency between the receptor binding model and the experimental data. Thus, the conjugate gradients method can be used to distinguish among receptor binding models. PMID- 15546493 TI - Females do not have more injury road accidents on Friday the 13th. AB - BACKGROUND: This study reinvestigated the recent finding that females - but not males - die in traffic accidents on Friday the 13th more often than on other Fridays (Nayha S: Traffic deaths and superstition on Friday the 13th. Am J Psychiatry 2002, 159: 2110-2111). The current study used matched setting and injury accident data base that is more numerous than fatality data. If such an effect would be caused by impaired psychic and psychomotor functioning due to more frequent anxiety among women, it should also appear in injury crashes. METHODS: We used the national Finnish road accident database for 1989-2002. To control seasonal variation, 21 Fridays the 13th were compared in a matched design to previous and following Fridays, excluding all holidays, on number of accidents, male/female responsibility for accidents, and the number of dead, injured and overall number of active participants (drivers, pedestrians and bicyclists) as a consequence of the accident. RESULTS: There were no significant differences in any examined aspect of road injury accidents among the three Fridays, either in females or males. Women were not overrepresented in crashes that occurred on Fridays 13th. CONCLUSION: There is no consistent evidence for females having more road traffic crashes on Fridays the 13th, based on deaths or road accident statistics. However, this does not imply a non-existent effect of superstition related anxiety on accident risk as no exposure-to-risk data are available. People who are anxious of "Black Friday" may stay home, or at least avoid driving a car. PMID- 15546492 TI - Metabolic scaling: consensus or controversy? AB - BACKGROUND: The relationship between body mass (M) and standard metabolic rate (B) among living organisms remains controversial, though it is widely accepted that in many cases B is approximately proportional to the three-quarters power of M. RESULTS: The biological significance of the straight-line plots obtained over wide ranges of species when B is plotted against log M remains a matter of debate. In this article we review the values ascribed to the gradients of such graphs (typically 0.75, according to the majority view), and we assess various attempts to explain the allometric power-law phenomenon, placing emphasis on the most recent publications. CONCLUSION: Although many of the models that have been advanced have significant attractions, none can be accepted without serious reservations, and the possibility that no one model can fit all cases has to be more seriously entertained. PMID- 15546494 TI - Myasthenia gravis and pregnancy: clinical implications and neonatal outcome. AB - BACKGROUND: The myasthenia gravis is twice as common in women as in men and frequently affects young women in the second and third decades of life, overlapping with the childbearing years. Generally, during pregnancy in one third of patients the disease exacerbates, whereas in two thirds it remains clinically unchanged. Complete remission can occur in some patients. METHODS: To describe the clinical course, delivery and neonatal outcome of 18 pregnant women with the diagnosis of myasthenia gravis. Retrospective chart review of pregnant patients with myasthenia gravis, followed at the National Institute of Perinatology in Mexico City over an 8-year period. Data was abstracted from the medical records on the clinical course during pregnancy, delivery and neonatal outcome. RESULTS: From January 1, 1996 to December 31, 2003 18 patients with myasthenia gravis were identified and included in the study. The mean +/- SD maternal age was 27.4 +/- 4.0 years. During pregnancy 2 women (11%) had an improvement in the clinical symptoms of myasthenia gravis, 7 women (39%) had clinical worsening of the condition of 9 other patients (50%) remained clinically unchanged. Nine patients delivered vaginally, 8 delivered by cesarean section and 1 pregnancy ended in fetal loss. Seventeen infants were born at mean +/- SD gestational age of 37.5 +/ 3.0 weeks and a mean birth weight of 2710 +/- 73 g. Only one infant presented with transient neonatal myasthenia gravis. No congenital anomalies were identified in any of the newborns. CONCLUSIONS: The clinical course of myasthenia gravis during pregnancy is variable, with a significant proportion of patients experiencing worsening of the clinical symptoms. However, neonatal transient myasthenia was uncommon in our patient population. PMID- 15546495 TI - Reserpine methonitrate, a novel quaternary analogue of reserpine augments urinary excretion of VMA and 5-HIAA without affecting HVA in rats. AB - BACKGROUND: Reserpine, an alkaloid from Rauwolfia serpentina was widely used for its antihypertensive action in the past. In later years, its use has been reduced because of precipitation of depression and extra pyramidal symptoms due to its central action. In the present investigation, reserpine methonitrate (RMN), a novel quaternary analogue of reserpine was synthesised and evaluated biochemically for its central and peripheral amine depleting actions in rats while its influence on the blood pressure was measured in anaesthetized rats in comparison with reserpine RESULTS: Reserpine treatment (5 mg/kg) produced a significant increase in the urinary excretion of VMA, 5-HIAA and HVA while RMN at doses of equal to and double the equimolar doses of reserpine (5 and 10 mg/kg) produced significant increase in VMA and 5-HIAA excretion without producing any effect on HVA excretion compared to control animals. Reserpine in the dose range of 0.5 to 15 microg/kg produced significant reduction in blood pressure compared to control. RMN was also found to produce significant decrease in blood pressure at doses of 10, 25 and 50 microg/kg body weight in comparison to control. The results indicated peripheral depletion of biogenic amines by RMN without affecting the central stores of the amines. CONCLUSIONS: The present study clearly indicated that the quaternization of reserpine restricts its transfer across the blood-brain barrier and could be the reason for its selective peripheral action. It is also clear that the hypotensive actions of RMN could be due to peripheral depletion of catecholamines. PMID- 15546496 TI - The use of microbubbles to target drug delivery. AB - Ultrasound-mediated microbubbles destruction has been proposed as an innovative method for noninvasive delivering of drugs and genes to different tissues. Microbubbles are used to carry a drug or gene until a specific area of interest is reached, and then ultrasound is used to burst the microbubbles, causing site specific delivery of the bioactive materials. Furthermore, the ability of albumin coated microbubbles to adhere to vascular regions with glycocalix damage or endothelial dysfunction is another possible mechanism to deliver drugs even in the absence of ultrasound. This review focuses on the characteristics of microbubbles that give them therapeutic properties and some important aspects of ultrasound parameters that are known to influence microbubble-mediated drug delivery. In addition, current studies involving this novel therapeutical application of microbubbles will be discussed. PMID- 15546497 TI - Spider silks: recombinant synthesis, assembly, spinning, and engineering of synthetic proteins. AB - Since thousands of years humans have utilized insect silks for their own benefit and comfort. The most famous example is the use of reeled silkworm silk from Bombyx mori to produce textiles. In contrast, despite the more promising properties of their silk, spiders have not been domesticated for large-scale or even industrial applications, since farming the spiders is not commercially viable due to their highly territorial and cannibalistic nature. Before spider silks can be copied or mimicked, not only the sequence of the underlying proteins but also their functions have to be resolved. Several attempts to recombinantly produce spider silks or spider silk mimics in various expression hosts have been reported previously. A new protein engineering approach, which combines synthetic repetitive silk sequences with authentic silk domains, reveals proteins that closely resemble silk proteins and that can be produced at high yields, which provides a basis for cost-efficient large scale production of spider silk-like proteins. PMID- 15546498 TI - Zinc/copper imbalance reflects immune dysfunction in human leishmaniasis: an ex vivo and in vitro study. AB - BACKGROUND: The process of elimination of intracellular pathogens, such as Leishmania, requires a Th1 type immune response, whereas a dominant Th2 response leads to exacerbated disease. Experimental human zinc deficiency decreases Th1 but not Th2 immune response. We investigated if zinc and copper levels differ in different clinical forms of leishmaniasis, and if these trace metals might be involved in the immune response towards the parasite. METHODS: Blood was collected from 31 patients with either localized cutaneous (LCL), mucosal (ML) or visceral (VL) leishmaniasis, as well as from 25 controls from endemic and non endemic areas. Anti-Leishmania humoral and cellular immune response were evaluated by quantifying specific plasma IgG, lymphoproliferation and cytokine production, respectively. Plasma levels of Cu and Zn were quantified by atomic absorption spectrophotometry. RESULTS: A significant decrease in plasma Zn was observed in all three patient groups (p < 0.01 for LCL and ML, p < 0.001 for VL), as compared to controls, but only VL (7/10) and ML (1/7) patients displayed overt Zn deficiency. Plasma Cu was increased in LCL and VL (p < 0.001) but not in ML, and was strongly correlated to anti-Leishmania IgG (Spearman r = 0.65, p = 0.0028). Cu/Zn ratios were highest in patients with deficient cellular (VL<LCL>ML) immune response. Ex vivo production of parasite-induced IFN-gamma was negatively correlated to plasma Cu levels in LCL (r = -0.57, p = 0.01). In vitro, increased Cu levels inhibited IFN gamma production. CONCLUSIONS: 1. Zn deficiency in VL and ML indicate possible therapeutic administration of Zn in these severe forms of leishmaniasis. 2. Plasma Cu positively correlates to humoral immune response across patient groups. 3. Environmentally or genetically determined increases in Cu levels might augment susceptibility to infection with intracellular pathogens, by causing a decrease in IFN-gamma production. PMID- 15546500 TI - Exposure to juvenile stress exacerbates the behavioural consequences of exposure to stress in the adult rat. AB - To examine the effects of exposure to post-weaning pre-puberty (juvenile) stress on the emotional and cognitive abilities in response to exposure to stress in adulthood, we first exposed rats to a platform stress at the age of 28 d. Two months later the rats were exposed to acute swim stress. Rats exposed to both stressors showed a higher level of anxiety (as measured both in open-field and startle response tests) than controls or rats exposed to either the juvenile or the adulthood stressor. In the Morris water-maze, rats that were exposed to both juvenile and adulthood stress performed better than the other groups. In a second experiment we verified that the effect of the juvenile stress was indeed age dependent. One group was exposed at the age of 26-28 d and again at the age of 60 d (juvenile + adulthood stress); the other group was exposed to the first stressor at the age of 60-62 d and to the second at the age of 90 d [adulthood (60) + adulthood (90) stress]. Juvenile + adulthood stress had a significantly greater effect than exposure to stress twice in adulthood, on anxiety level and on the performance in the water-maze. Finally, in a third experiment we found that the juvenile+adulthood stress group swam faster and tended to explore the central area more than the other groups--a finding that could explain their better performance on the first trial of the spatial task. These results indicate that an exposure to a relatively brief juvenile stressful experience has profound and long-lasting effects on the ability to cope with stress in adulthood. PMID- 15546499 TI - Young age: an independent risk factor for disease-free survival in women with operable breast cancer. AB - BACKGROUND: The incidence of breast cancer in young women (age < 35) is low. The biology of the disease in this age group is poorly understood, and there are conflicting data regarding the prognosis for these women compared to older patients. METHODS: We retrospectively analyzed 2040 consecutive primary invasive breast cancer patients who underwent surgical procedures at our institution between 1990 and 1999. The younger age group was defined as patients aged <35 years at the time of diagnosis. The clinicopathological characteristics and treatment outcomes were compared between younger and older age groups. RESULTS: A total of 256 (12.5%) patients were aged <35. There was a significantly higher incidence of nuclear grade 3 and medullary histological-type tumors in younger patients compared to older patients. Axillary lymph node status, T stage, histological grade, c-erbB2 expression and estrogen receptor status did not differ significantly between the two age groups. Younger patients had a greater probability of recurrence and death at all time periods. Although there was no significant difference in disease-free survival between the two age groups in lymph node-negative patients, the younger group showed worse prognosis among lymph node-positive patients (p < 0.001). In multivariate analysis, young age remained a significant predictor of recurrence (p = 0.010). CONCLUSION: Young age (<35) is an independent risk factor for relapse in operable breast cancer patients. PMID- 15546501 TI - The role of thymidine phosphorylase, an angiogenic enzyme, in tumor progression. AB - Thymidine phosphorylase (TP), an enzyme involved in pyrimidine metabolism, is identical with an angiogenic factor, platelet-derived endothelial cell growth factor (PD-ECGF). TP is overexpressed in various tumors and plays an important role in angiogenesis, tumor growth, invasion and metastasis. The enzymatic activity of TP is required for the angiogenic effect of TP. A novel, specific TP inhibitor, TPI, inhibits angiogenesis induced by overexpression of TP in KB/TP cells (human KB epidermoid carcinoma cells transfected with TP cDNA), as well as the growth and metastasis of KB/TP cells in vivo. 2-deoxy-D-ribose, the degradation product of thymidine generated by TP activity, has both angiogenic and chemotactic activity. Both 2-deoxy-D-ribose and TP inhibit a hypoxia-induced apoptotic pathway. These findings suggest that 2-deoxy-D-ribose is a downstream mediator of TP function. 2-deoxy-L-ribose, a stereoisomer of 2-deoxy-D-ribose, inhibits the promotion of angiogenesis, tumor growth and metastasis by TP. Although the mechanism of the action of 2-deoxy-D-ribose is still unknown, 2 deoxy-L-ribose may inhibit the physiological activities of 2-deoxy-D-ribose, and consequently those of TP. Inhibition of TP activity and function appears to be a promising approach for the chemotherapy of various tumors. PMID- 15546502 TI - Interferon-beta gene therapy for cancer: basic research to clinical application. AB - Interferon-beta gene therapy for cancer is the first such protocol developed in Japan. Here we describe the development process of our interferon-beta gene therapy from basic research to clinical application. Interestingly, the biological and biochemical characteristics of interferon-beta gene therapy through transfer of the interferon-beta gene into tumor cells by means of cationic liposomes differed from those of conventional interferon-beta protein therapy. Interferon-beta gene transfer could induce apoptosis in interferon-beta protein-resistant tumor cells, such as glioma, melanoma, and renal cell carcinoma. Induction of apoptosis was related to the level of intracellular mRNA of interferon-beta, prolongation of the phosphorylation time of molecules in the interferon-beta signal transduction pathway, such as JAK1, Trk2, and STAT1, and activation of DNase gamma. In our preclinical study we developed lyophilized cationic liposomes containing interferon-beta gene (gene drug) for clinical use and confirmed their safety. Thereafter, we performed a pilot clinical trial in patients with malignant glioma and confirmed the safety and effectiveness of this interferon-beta gene therapy. In this review we also comment on the status of gene therapy regulation in Japan. Interferon-beta gene therapy is expected to become widely available for clinical use in cancer patients, and this new strategy might be extended to molecular targeting therapy, or used in combination with cell therapy or other therapies. PMID- 15546503 TI - Role of BRCA1 and BRCA2 as regulators of DNA repair, transcription, and cell cycle in response to DNA damage. AB - BRCA1 (BReast-CAncer susceptibility gene 1) and BRCA2 are tumor suppressor genes, the mutant phenotypes of which predispose to breast and ovarian cancers. Intensive research has shown that BRCA proteins are involved in a multitude of pivotal cellular processes. In particular, both genes contribute to DNA repair and transcriptional regulation in response to DNA damage. Recent studies suggest that BRCA proteins are required for maintenance of chromosomal stability, thereby protecting the genome from damage. New data also show that BRCAs transcriptionally regulate some genes involved in DNA repair, the cell cycle, and apoptosis. Many of these functions are mediated by a large number of cellular proteins that interact with BRCAs. The functions of BRCA proteins are also linked to distinct and specific phosphorylation events; however, the extent to which phosphorylation-activated molecular pathways contribute to tumor suppressor activity remains unclear. Finally, the reasons why mutations in BRCA genes lead to the development of breast and ovarian cancers are not clearly understood. Elucidation of the precise molecular functions of BRCAs is expected to improve our understanding of hereditary as well as sporadic mammary carcinogenesis. PMID- 15546504 TI - Eradication of Helicobacter pylori induces apoptosis and inhibits proliferation of heterotopic proliferative glands in infected Mongolian gerbils. AB - Mongolian gerbils infected with Helicobacter pylori (H. pylori ) develop heterotopic proliferative glands (HPGs) in the glandular stomach submucosa. To investigate the effects of H. pylori eradication on cell turnover in HPGs, three antibiotics, lansoprazole, amoxicillin and clarithromycin, were administered at 50 or 75 weeks after inoculation of H. pylori, and the stomachs were excised for histological examination at 1, 2, 4, 8 or 25 weeks thereafter. The HPGs were classified into gastric type (G-type) and others (GI + I-type), which included both pure intestinal (I-type) and gastric-and-intestinal mixed type (GI-type). Apoptosis and cell proliferation were evaluated by means of TUNEL assay and BrdU labeling, respectively. At 8 weeks post-eradication, apoptotic indices were significantly increased in the eradication group (G-type: 2.5%; GI + I-type: 7.2%) compared to the non-eradication group (G-type: 0.6%; GI + I-type: 2.1%: P < 0.01), while BrdU labeling indices were significantly decreased (G-type: 1.9%; GI + I-type: 6.8% as compared with 4.3% and 13.2%, respectively, P < 0.01 for both). At 25 weeks, the apoptotic indices were similarly higher [G-type: 0.4 (eradication group) vs. 0.2% (non-eradication group); GI + I-type: 5.8 vs. 1.1%, both P < 0.01], and the BrdU labeling indices (G-type: 0.8 vs. 2.2%, P < 0.01; GI + I-type: 5.1 vs. 11%, P < 0.05) continued to be lower in HPGs. Furthermore, there were highly significant reductions in the areas of HPGs at 8 and 25 weeks post-eradication. These findings demonstrated that eradication results in apoptosis and reduction of proliferation of HPGs in H. pylori-infected gerbils, these lesions thus being apparently reversible through regulation of cell kinetics. PMID- 15546505 TI - Transforming growth factor-beta 1-induced apoptosis is blocked by beta 1-integrin mediated mitogen-activated protein kinase activation in human hepatoma cells. AB - Growth factors and extracellular matrices cooperatively regulate cellular behavior. However, the interactions between transforming growth factor-beta 1 (TGF-beta 1) and integrins in hepatic cells are not fully understood. We investigated the effects of beta 1-integrin on TGF-beta 1-regulated growth of hepatoma cells. Human hepatoma cell lines HepG2, Huh7, and Hep3B were stably transfected with beta 1-integrin, and the parental, and mock- and beta 1-integrin transfected hepatoma cells were treated with TGF-beta 1. Modulation of apoptosis and pathways involved in the process were investigated. TGF-beta 1 suppressed the growth of hepatoma cells, and apoptosis was observed in Hep3B and Huh7. Hepatoma cells transfected with beta 1-integrin were protected from TGF-beta 1-induced apoptosis. Mitogen-activated protein (MAP) kinase inhibitors, PD98059, SB203580, and SP600125, abolished this protective effect of beta 1-integrin, but herbimycin A and wortmannin were ineffective. Hepatoma cells overexpressing beta 1-integrin showed increased activities of MAP kinases, and TGF-beta 1 induced sustained activation of MAP kinases in these cells, but only transient activation in mock transfected cells. These data suggest that MAP kinases activated by beta 1 integrin provide a strong anti-apoptotic signal during TGF-beta 1-induced apoptosis in human hepatoma cells. Therefore beta 1-integrin-mediated signals may contribute to the development and progression of hepatocellular carcinoma. PMID- 15546506 TI - mRNA expression level of estrogen-inducible gene, alpha 1-antichymotrypsin, is a predictor of early tumor recurrence in patients with invasive breast cancers. AB - We previously identified 18 genes that correlated with ER positivity by adapter tagged competitive-PCR analysis of 2412 genes in human breast cancer tissues. The aim of the present study was to determine the prognostic significance of these genes. mRNA expression levels of 12 of the above 18 genes were quantified in breast cancer tissues by real-time PCR assay, and their association with patients' prognosis (n = 110) was studied according to hormone receptor (HR) status. In addition, the genes found to influence prognosis were further investigated to examine whether their mRNA expression could be induced by estrogen in MCF-7 cells in vitro. Of the 12 genes, mRNA expression levels of two [alpha 1-antichymotrypsin (ACT) and stanniocalcin 2 (STC2)] were significantly (P = 0.002 and P = 0.007, respectively) associated with good prognosis in HR positive (ER and/or PR positive) breast cancer patients treated with adjuvant hormone therapy. Multivariate analysis showed that ACT mRNA level, but not STC2 mRNA level, in HR-positive patients, was a significant prognostic factor (P = 0.042), which was independent of tumor size and lymph node metastases. On the other hand, mRNA expressions of ACT and STC2 were not significantly associated with prognosis in HR-negative patients. Estradiol treatment resulted in a significant increase in the mRNA levels of both ACT and STC2 in MCF-7 cells. The mRNA level of ACT, which is an estrogen-inducible gene, is a significant predictor of good prognosis in HR-positive, but not HR-negative, patients with breast cancers. Since HR-positive patients were treated with adjuvant hormone therapy, we suggest that ACT mRNA level could potentially be used as a predictor of response to hormone therapy, rather than a prognostic factor (predictor of metastatic potential). PMID- 15546507 TI - Novel models for human scirrhous gastric carcinoma in vivo. AB - Human scirrhous gastric carcinoma, a diffusely infiltrating type of poorly differentiated gastric carcinoma also known as linitis plastica type carcinoma, is characterized by cancer cell infiltration and proliferation accompanied with extensive stromal fibrosis. We established two new gastric cancer cell lines, designated OUCM-8 and OCUM-11, which developed the characteristic biology of scirrhous gastric carcinoma upon orthotopic implantation in mice. Involvement of lymph nodes and liver metastasis was also found in both orthotopic models. Histologically, these orthotopic models showed proliferation with extensive fibrosis, resembling human scirrhous gastric cancer. Both cell lines were derived from ascites of patients with scirrhous gastric cancer. The growth of OCUM-8 and OCUM-11 cells following the addition of KGF, FGF, and EGF was increased significantly relative to untreated cells. An increase in the number of attached and spreading cells occurred following the addition of TGF-beta 1 in both cell lines. OCUM-11 cells showed microsatellite instability. Although subcutaneous scirrhous gastric cancer cells show medullary growth, most in vivo studies of scirrhous gastric cancer have used xenografted tumors implanted subcutaneously. Only in a few cases was it confirmed that these scirrhous gastric cancer cell lines retained the original histologic characteristics. Our orthotopic models should contribute to the elucidation of disease progression in situ and to the development of therapy for scirrhous gastric cancer. PMID- 15546508 TI - Non-steroidal anti-inflammatory drugs inhibit cellular proliferation and upregulate cyclooxygenase-2 protein expression in endometrial cancer cells. AB - We determined the effects of several non-steroidal anti-inflammatory drugs (NSAIDs), aspirin (acetylsalicylic acid, ASA), indomethacin and a cyclooxygenase 2 (COX-2)-selective inhibitor (NS398), on cellular proliferation and regulation of COX-2 protein expression in endometrial cancer cells in vitro, and investigated their modes of action. All three NSAIDs markedly inhibited the proliferation of Ishikawa, HEC-1A and AN3CA endometrial cancer cell lines in a time- and concentration-dependent manner. ASA and indomethacin triggered apoptosis in cells of all three lines through release of cytosolic cytochrome c, activation of caspase-9 and-3, and cleavage of poly(ADP-ribose) polymerase (PARP), but NS398 induced minimal apoptosis only in Ishikawa cells. ASA altered the cell cycle distribution, with G2/M phase accumulation of cells, and induced overexpression of Ki-67 protein. Both ASA and indomethacin reduced the protein levels of Bcl-2 and Bcl-xl, but upregulated those of Bax and Bcl-xs. COX-2 protein expression and PGE(2) production were upregulated by ASA and indomethacin in all three cell lines. However, NS398 did not alter COX-2 protein expression or PGE(2) production in these cells. These results indicate that NSAIDs inhibit proliferation of endometrial cancer cells independently of the reduction of COX-2 protein expression. A cytochrome c-dependent apoptotic pathway and/or cell cycle arrest may contribute to the inhibitory effects of these NSAIDs. PMID- 15546509 TI - Methylenetetrahydrofolate reductase C677T and A1298C polymorphisms and colorectal cancer: the Fukuoka Colorectal Cancer Study. AB - Methylenetetrahydrofolate reductase (MTHFR) is a key enzyme regulating folate metabolism, which affects DNA synthesis and methylation. This study investigated the relation of MTHFR C677T and A1298C polymorphisms to colorectal cancer in a case-control study in Fukuoka, Japan. The subjects comprised 685 incident cases of histologically confirmed colorectal adenocarcinomas and 778 community controls selected randomly in the study area. The genotype was determined by the PCR-RFLP method using genomic DNA extracted from buffy coat. Alcohol use was ascertained by in-person interview. Statistical adjustment was made for gender, age class, area, and alcohol use. The MTHFR 677TT genotype was associated with a statistically significant decrease in the risk with an adjusted odds ratio of 0.69 (95% confidence interval 0.51-0.93) compared with the 677CC and 677CT combined, and the decrease was most evident in individuals with no alcohol consumption. While the A1298C polymorphism showed no measurable association with the overall risk of colorectal cancer, the 1298CC genotype was associated with a statistically significant increase in the risk when alcohol consumption was high, and was also associated with an approximately 2-fold increase in the risk of each of proximal and distal colon cancer. The findings add to evidence that individuals with the MTHFR 677TT genotype have a decreased risk of colorectal cancer in the absence of folate depletion, suggesting a protective role of folate by ensuring a sufficient thymidylate pool for DNA synthesis. Because very few individuals had the 1298CC genotype, the findings regarding the A1298C polymorphism need careful interpretation and confirmation in larger studies. PMID- 15546510 TI - Immunity against mouse thymus-leukemia antigen (TL) protects against development of lymphomas induced by a chemical carcinogen, N-butyl-N-nitrosourea. AB - Mouse thymus-leukemia antigens (TL) are aberrantly expressed on T lymphomas in C57BL/6 (B6) and C3H/He (C3H) mice, while they are not expressed on normal T lymphocytes in these strains. When N-butyl-N-nitrosourea (NBU), a chemical carcinogen, was administered orally to B6 and C3H strains, lymphoma development was slower than in T3(b)-TL gene-transduced counterpart strains expressing TL ubiquitously as self-antigens, suggesting that anti-TL immunity may play a protective role. In addition, the development of lymphomas was slightly slower in C3H than in B6, which seems to be in accordance with the results of skin graft experiments indicating that both cellular and humoral immunities against TL were stronger in C3H than B6 mice. The interesting finding that B lymphomas derived from a T3(b)-TL transgenic strain (C3H background) expressing a very high level of TL were rejected in C3H, but not in H-2K(b) transgenic mice (C3H background), raises the possibility that TL-specific effector T cell populations are eliminated and/or energized to a certain extent by interacting with H-2K(b) molecules. PMID- 15546516 TI - [Impact of metabolic syndrome in the control of blood pressure and dyslipemia]. AB - BACKGROUND AND OBJECTIVE: The objective of the study was to assess the influence of metabolic syndrome (MS) in the control of blood pressure (BP) and dyslipemia. PATIENTS AND METHOD: A cross sectional study was performed with 1,320 (634 M and 686 F), 40.1 (13.3) years-old, BMI 29.8 (4.7) hypertensive non-diabetic patients. MS was diagnosed according to NCEP-ATP-III guidelines. Blood pressure control goal was BP < 140/90 mmHg. Coronary risk (CR) was calculated according to Framingham (low < 10%, intermediate 10-20% and high > 20% at 10 years). Goals of C-LDL levels were those of NCEP-ATP-III. RESULTS: 461 (35%) patients had MS and the remaining 859 became controls. Patients with MS had higher initial levels of hypertension and were receiving more antihypertensive drugs: 2.1 [1.3] vs. 1.7 [1.3]; p < 0.001), yet the average systolic and diastolic BP achieved and the degree of control was similar in both groups 53% vs. 52%; (p = ns). Patients with MS had higher CR at ten years than controls (10.7 [8.3] vs. 7.9 [6.8], p < 0.001) but achieved the C-LDL goals at fewer proportions than controls (57% vs. 74%; p < 0.001). In a regression analysis, patients with MS had 26% less probabilities of achieving both goals (p < 0.001). CONCLUSIONS: Hypertensive patients with MS have higher CR, and need more antihypertensive drugs to achieve the same BP goals. Yet it is more difficult for them to achieve LDL cholesterol goals. Patients with MS remain a target for cardiovascular prevention. PMID- 15546515 TI - Virtual outreach: a randomised controlled trial and economic evaluation of joint teleconferenced medical consultations. AB - OBJECTIVES: To test the hypotheses that virtual outreach would reduce offers of hospital follow-up appointments and reduce numbers of medical interventions and investigations, reduce numbers of contacts with the health care system, have a positive impact on patient satisfaction and enablement, and lead to improvements in patient health status. To perform an economic evaluation of virtual outreach. DESIGN: A randomised controlled trial comparing joint teleconsultations between GPs, specialists and patients with standard outpatient referral. It was accompanied by an economic evaluation. SETTING: The trial was centred on the Royal Free Hampstead NHS Trust, London, and the Royal Shrewsbury Hospital Trust in Shropshire. The project teams recruited and trained a total of 134 GPs from 29 practices and 20 consultant specialists. PARTICIPANTS: In total, 3170 patients were referred, of whom 2094 consented to participate in the study and were eligible for inclusion. In all, 1051 patients were randomised to the virtual outreach group and 1043 to standard outpatient appointments. The patients were followed 6 months after their index consultation. INTERVENTIONS: Patients randomised to virtual outreach underwent a joint teleconsultation, in which they attended the general practice surgery where they and their GP consulted with a hospital specialist via a videolink between the hospital and the practice. MAIN OUTCOME MEASURES: Outcome measures included offers of follow-up outpatient appointments, numbers of tests, investigations, procedures, treatments and contacts with primary and secondary care, patient satisfaction (Ware Specific Visit Questionnaire), enablement (Patient Enablement Instrument) and quality of life (Short Form-12 and Child Health Questionnaire). An economic evaluation of the costs and consequences of the intervention was undertaken. Sensitivity analysis was used to test the robustness of the results. RESULTS: Patients in the virtual outreach group were more likely to be offered a follow-up appointment. Significant differences in effects were observed between the two sites and across different specialities. Virtual outreach increased the offers of follow-up appointments more in Shrewsbury than in London, and more in ENT and orthopaedics than in the other specialities. Fewer tests and investigations were ordered in the virtual outreach group, by an average of 0.79 per patient. In the 6-month period following the index consultation, there were no significant differences overall in number of contacts with general practice, outpatient visits, accident and emergency contacts, inpatient stays, day surgery and inpatient procedures or prescriptions between the randomised groups. Tests of interaction indicated that virtual outreach decreased the number of tests and investigations, particularly in patients referred to gastroenterology, and increased the number of outpatient visits, particularly in those referred to orthopaedics. Patient satisfaction was greater after a virtual outreach consultation than after a standard outpatient consultation, with no heterogeneity between specialities or sites. However, patient enablement after the index consultation, and the physical and psychological scores of the Short Form-12 for adults and the scores on the Child Health Questionnaire for children under 16, did not differ between the randomised groups at 6 months' follow-up. NHS costs over 6 months were greater for the virtual outreach consultations than for conventional outpatients, pound 724 and pound 625 per patient, respectively. The index consultation accounted for this excess. Cost and time savings to patients were found. Estimated productivity losses were also less in the virtual outreach group. CONCLUSIONS: Virtual outreach consultations result in significantly higher levels of patient satisfaction than standard outpatient appointments and lead to substantial reductions in numbers of tests and investigations, but they are variably associated with increased rates of offer of follow-up according to speciality and site. Changes in costs and technological advances may improve the relative position of virtual consultations in future. The extent to which virtual outreach is implemented will probably be dependent on factors such as patient demand, costs, and the attitudes of staff working in general practice and hospital settings. Further research could involve long-term follow-up of patients in the virtual outreach trial to determine downstream outcomes and costs; further study into the effectiveness and costs of virtual outreach used for follow-up appointments, rather than first-time referrals; and whether the costs of virtual outreach could be substantially reduced without adversely affecting the quality of the consultation if nurses or other members of the primary care team were to undertake the hosting of the joint teleconsultations in place of the GP. Qualitative work into the attitudes of the patients, GPs and hospital specialists would also be valuable. PMID- 15546517 TI - [Physical activity and quality of life in older adults in Spain]. AB - BACKGROUND AND OBJECTIVE: This study examined the relationship between leisure time physical activity (LTPA) and health-related quality of life (HRQL) in the older adult population of Spain. SUBJECTS AND METHOD: Household cross-sectional survey on 3,066 subjects representatives of the non-institutionalized Spanish population aged 60 years and over. Data on LTPA was obtained with a structured questionnaire and HRQL was measured with the SF-36 instrument. Analyses were done through linear regression, where the dependent variable was each of the eight scales of the SF-36 and the main independent variable was LTPA. Analyses were adjusted for sociodemographic and social network variables, health habits, health services use, and chronic diseases. RESULTS: A total of 42.7% subjects had a sedentary activity, 54.2% light LTPA and 3% moderate/intense LTPA. As compared with sedentary activity, light LTPA was associated with a higher score in all SF 36 scales, except for the physical role and emotional role, among men and women. For subjects with light LTPA the increase in score was over 3 points in most SF scales, which is usually considered as a clinically relevant change in HRQL. Results did not vary materially by age, level of education, obesity or chronic disease. The higher LTPA, the better HRQL (p for linear trend < 0.05 in most scales of the SF-36 questionnaire). CONCLUSIONS: Light LTPA is associated with better HRQL than sedentary activity. Because this association did not change with age, level of education, obesity or chronic disease, it is suggested that most older adults could improve their HRQL with, at least, a light LTPA. PMID- 15546518 TI - [Study of predictive factors of severe digestive lesions due to caustics ingestion]. AB - OBJECTIVE: Our purpose was to analyze the predictive factors of severe upper gastrointestinal injury by caustic substances in adult patients. PATIENTS AND METHOD: Retrospective study between February 1995 and February 2001 of adult patients who underwent an urgent upper endoscopy due to caustic ingestion. Endoscopic caustic ingestion criteria by Zargar et al were used to determine the degree of injury. We performed a univariate study of factors associated with sever digestive injury and, lately, a logistic regression analysis of predictive factors. Sensitivity, specificity, positive and negative predictive values of these factors were calculated. RESULTS: One hundred and fifty nine patients were included in the study, whose mean age was 48.9 (20.1) years and 49.7% were men. The more frequent caustic ingested was lye (47.8%). A severe caustic injury was found in urgent upper endoscopy in 18.4% of patients, which was located in esophagus in 14.6%, stomach in 8.2% and duodenum in 0.6% of cases. Male sex, voluntary ingestion, oropharingeal lesions, significant clinical symptoms and dishwasher and detergents ingestion were associated with severe upper gastrointestinal tract (GIT) injury. Voluntary ingestion, oropharingeal lesions and significant clinical symptoms at admission were independent predictive factors of severe GIT injury. The existence of one of these factors had an 89.7% of sensitivity while two or more displayed a specificity of 91%. CONCLUSIONS: Clinical and exploratory data may determine, before upper endoscopic procedure, the probability of severe GIT injury by caustic ingestion. Therefore, these data could play a significant role in the diagnostic, prognostic and therapeutic management of caustic ingestion. PMID- 15546519 TI - [Appropriateness of the utilization of an emergency department]. AB - BACKGROUND AND OBJECTIVE: The increase in the demand of clinical attention in emergency departments justifies the need to evaluate its adequacy to the population health needs requiring emergency attention. The objective of this study is to evaluate the adequacy of patients' visits to the emergency departments of a third level hospital. MATERIAL AND METHOD: Descriptive study with analytic components. Random sample: 499 medical reports of patients treated in the Emergency Department. The HUAP (Appropriateness Hospital Emergencies Protocol) protocol was applied. We carried out a descriptive analysis of the sample, a relation analysis and finally we carried out a multiple logistic regression analysis. RESULTS: 30.7% visits were inadequate. The higher percentage of inappropriateness was found in subjects younger than 40 years (44%); in those without chronic pathology (53.3%) and in those who came spontaneously (88.4%). Half of the appropriate visits owed, exclusively, to achieving "diagnostic tests" criteria. CONCLUSIONS: The Appropriateness Hospital Emergencies Protocol allowed to identify two population groups. It is necessary to implement strategies to change the health services offer, adapted to the differences found and taking into account the increase in the appropriate demand. PMID- 15546520 TI - [Inappropriate emergencies or insufficient offer?]. PMID- 15546521 TI - [Short stay units: appropiateness of the hospital admittance]. PMID- 15546522 TI - [Restricted use of personal data in health practice and research: a case of a sentence involving the registry of HIV infection]. PMID- 15546523 TI - [TNF-alpha inhibitors in inflammatory bowel disease]. AB - Inflammatory bowel disease (IBD) is a chronic immunoinflammatory response to an stimulus that activates a chain of cellular mediators causing intestinal damage. One of the most well recognized proinflammatory mediators involved in the pathogenesis of IBD is tumor necrosis factor alpha (TNFalpha). The treatment of IBD has advanced in parallel to the improvement of the knowledge of its physiopathology, leading to the development of biological therapies. An example of this kind of treatment is the use of substances that antagonize TNFalpha, such as monoclonal antibodies infliximab, adalimumab, natalizumab, etanercept or onercept, with infliximab being the unique approved for use in IBD. Several studies have demonstrated that inhibition of TNFalpha is useful in the treatment of Crohn's disease (CD). In CD, infliximab induces the remission of relapses which are refractory to the conventional treatment, prevents more relapses and induces a closure of enterocutaneous and perianal fistula that do not respond to first line treatment. However, infliximab is not useful in ulcerative colitis. Infliximab treatment has some drawbacks, such as the development of anti infliximab antibodies, which cause a loss of efficacy of the treatment and hypersensitivity reactions. Other reported adverse effects of infliximab are the development of autoimmunity, such as that related with antinuclear or anti-DNA antibodies, or the reactivation of infections such as tuberculosis. In fact, a screening for tuberculosis is necessary before administration of infliximab. To reduce the adverse effects due to infliximab immunogenicity, several trials with humanized or completely human agents, such as adalimumab or onercept, are under way. Until the precise stimulus that triggers IBD is identified, biological therapies have a great future and the selective antagonism of TNFalpha is already a reality. PMID- 15546524 TI - [Negative D-dimer in patients with venous thromboembolic disease]. PMID- 15546525 TI - [Sentinel lymph node biopsy in cutaneous melanoma: a study of 45 cases in a series of 142 melanomas]. PMID- 15546526 TI - [Autochthonous amebiasis in Spain. Endemic or emergent disease]. PMID- 15546527 TI - [Mycoplasma pneumoniae pneumonia associated with hemolytic anemia and Stevens Johnson syndrome]. PMID- 15546528 TI - [Blister reaction after the administration of interleukin-2]. PMID- 15546529 TI - [Intermittent and subacute intestinal obstruction due to giant lipoma of the colon]. PMID- 15546530 TI - [Digoxin toxicity secondary to inhibition of digoxin intestinal metabolism in a patient receiving clarithromycin]. PMID- 15546531 TI - [New strategies of cardiovascular prevention: the importance of consensus]. PMID- 15546532 TI - [Inactivated polio vaccine]. PMID- 15546535 TI - [Hypertension and compliance with therapy]. PMID- 15546536 TI - [Effectiveness of an intervention to provide information to patients with hypertension as short text messages and reminders sent to their mobile phone (HTA Alert)]. AB - OBJECTIVE: To analyze the effect of an intervention to provide information with mobile phone text messages to patients with hypertension on compliance with therapy for hypertension. DESIGN: Comparative, controlled, multicenter, randomized cluster study. SETTING: 26 primary care health centers in Spain. PARTICIPANTS: 26 researchers were randomized to a control group or an intervention group (52 patients each, for a total of 104 patients). All patients were receiving monotherapy for uncontrolled hypertension. INTERVENTION: Patients in the control group received their physician's usual interventions. Patients in the intervention group received messages and reminders sent to their mobile phones 2 days per week during 4 months. MAIN OUTCOME MEASURES: Tablets were counted and blood pressure was measured at the start of the study and 1, 3, and 6 months later. The percentage of compliers, mean percentage of compliance and degree of control of hypertension were compared. The reduction in absolute and relative risk was calculated, as was the number of individuals needed to treat to avoid noncompliance. RESULTS: The results were evaluated for a total of 67 individuals (34 in the intervention group and 33 in the control group). The rate of compliance was 85.1% (CI, 74.9%-95.3%) overall, 85.7% (CI, 70.5%-100.9%) in the control group and 84.4% in the intervention group (CI, 70.7%-95.3%) (P=NS). Mean percentage compliance was 90.2%+/-16.3% overall, 88.1%+/-20.8% in the control group and 91.9%+/-11.6% in the intervention group (P=NS). The percentage of patients whose hypertension was controlled at the end of the study was 51.5% (CI, 34.4%-68.6%) in the control group and 64.7% (CI, 48.6%-80.8%) in the intervention group (P=NS). CONCLUSIONS: The telephone messaging intervention with alerts and reminders sent to mobile phones did not improve compliance with therapy in patients with hypertension. PMID- 15546537 TI - [Non-compliance with prescribed medication]. PMID- 15546538 TI - [Teaching of basic heart-lung reanimation to the general public]. AB - OBJECTIVE: To analyse the efficacy of training in basic paediatric and adult heart-lung reanimation by means of theoretical/practical courses for the general public. DESIGN: Prospective and observational study. SETTING: Chamberi Municipal Health Centre, Madrid. PARTICIPANTS: 360 people aged from 13 to 75. INTERVENTIONS: From 1999 to 2003, 381 people took 26 courses in prevention of accidents and response to them. Theoretical classes in prevention of cardiac/respiratory arrest and in basic heart-lung reanimation (HLR) and practical classes in HLR were given to groups of 5 or 6 students, using dummies of babies, children, and adults. MAIN MEASUREMENTS: At the start and end of the course, 17.2% of the participants were assessed for their theoretical knowledge; and 94.5% had a practical assessment of their HLR skills. At the end of the course all students filled in an anonymous satisfaction questionnaire. RESULTS: In the initial theoretical assessment, the mean score was 3.5+/-1.5 (out of a maximum of 10); in the final assessment, 7.7+/-1.7 (P<.01). 87.2% of participants achieved sufficient practical skills. In the questionnaire, the participants thought the course was very useful for daily life and felt more able to take action. CONCLUSIONS: The basic HLR courses for the general public provide useful theoretical and practical training and manage to improve peoples ability to react correctly and so resolve emergency situations. PMID- 15546539 TI - [Characteristics of oral anti-coagulation treatment in high-risk chronic auricular fibrillation]. AB - OBJECTIVE: To calculate the prevalence of chronic auricular fibrillation (CAF), how much of it is considered high-risk (CAFhr) and the degree of coverage with oral anti-coagulation treatment (OAT). DESIGN: Multi-centre descriptive study. SETTING: 9 health districts. The "Terres de l'Ebre" Primary Care Service. PARTICIPANTS: Randomised sample of 375 patients with CAF, of whom 150 met the criteria of CAFh-r during 2002. MAIN MEASUREMENTS: Profile of patients with CAFh r; coverage with OAT; place of follow-up; presence of vascular complications, and the time relationships between the diagnosis of CAF, vascular complications and the start of OAT. All the INR determinations taken from the patients included in the study were used. RESULTS: There was 2.2% prevalence of CAF (95% CI, 1.4-3.3). 40% of CAF had criteria of CAFh-r. 74.2% were treated with OAT. In 41.7% the diagnosis of CAF coincided with the incidence of some vascular complication. There were no differences between the overall results of the INR obtained in hospital and in PC. The expected efficacy of OAT for thromboembolism prevention in our high-risk sample was 61.12%. CONCLUSIONS: 40% of the CAF are high-risk. In over a third of patients OAT was indicated after a vascular complication linked to an unknown CAF. The INR between 2-3 is similar in PC centres and the corresponding haematology service. PMID- 15546540 TI - [Women who are partners of a man infected by HIV: description of their characteristics and appraisal of risk]. AB - OBJECTIVE: To describe the situations of risk and the prevalence of HIV in women with a heterosexual partner infected by HIV. DESIGN: Cross-sectional descriptive study. SETTING: Out-patient HIV diagnosis centre in Madrid. PATIENTS: 229 women seen for the first time between 1993 and 2002 because they had a stable heterosexual partner diagnosed with HIV, and who were exposed to no other risk. MAIN MEASUREMENTS: Social and personal details, reproduction history, sexual conduct, clinical, and analytic data of both partners. RESULTS: 66% of couples had maintained sexual relations for over a year. Women were on average younger (29.6 years old) than the men. 29% of the women had children and 5.2% were pregnant. 82% of men had injected drugs, but only 13% still did. 73% had been diagnosed with HIV for over 6 months, 16% had AIDS criteria, and 35% were taking retroviral treatment. 60% of the men who knew of their infection had systematically avoided sex without condoms, against 33% of those who did not know they were infected (P<.001). 19% had had accidents in use of the condom. HIV seroprevalence in the women was 6.1% (95% CI, 3.5%-10.3%). CONCLUSIONS: Health care of people with HIV must include care of their sexual partner, involving information, psychological support, preventive and reproductive advice, as well as diagnosis of HIV or other sexually transmitted diseases. PMID- 15546541 TI - [Spanish adaptation of the European Guide to Cardiovascular Prevention (I)]. PMID- 15546542 TI - [Family medicine as an academic discipline and primary care as a learning environment]. PMID- 15546543 TI - [Family doctors faced with sexual assaults]. PMID- 15546544 TI - [Situation of ongoing training and research in the Almeria primary care district in the views of professionals]. PMID- 15546545 TI - [Interferon and depression. Concerning a case]. PMID- 15546546 TI - [Effects on hospital prescription of the introduction of a form proposed by a primary care team]. PMID- 15546547 TI - [Self-medication in Spain.what can we do?]. PMID- 15546548 TI - [Specific treatment for arthrosis: glucosamine sulphate]. PMID- 15546549 TI - Why should the VA continue academic affiliations? AB - Academic affiliations between VA medical centers and medical schools have made an important contribution to the healthcare of veterans and the education of physicians over the last 60 years. The rationale for these affiliations and their establishment following World War II is discussed. The circumstances leading to affiliations and the benefit they afford veterans remain pertinent today. PMID- 15546550 TI - Prevalence of malignancy within cytologically indeterminate thyroid nodules. AB - BACKGROUND: The optimal management of cytologically indeterminate thyroid nodules is controversial given the variable malignancy rates reported in this patient population. We examined the prevalence of malignancy within cytologically indeterminate follicular thyroid lesions in an attempt to predict malignancy based on cytologic features. METHODS: Cytopathology reports obtained after fine needle aspiration biopsy (FNAB) examination of indeterminate follicular thyroid lesions were examined over a 4-year period. The prevalence of malignancy on final histology was determined in 4 indeterminate cytologic categories. RESULTS: A total of 107 records were available (91 women, 16 men). The mean patient age was 45.4 +/- 16 years. Forty-eight patients (45%) underwent surgery and had histopathologic diagnosis, while 57 patients did not have surgery. The prevalence of malignancy in patients who underwent thyroidectomy was 42% (20 of 48). CONCLUSIONS: The high prevalence of malignancy within indeterminate follicular lesions may necessitate thyroidectomy for patients with indeterminate follicular lesions on FNAB examination. PMID- 15546551 TI - Use of a predictive equation for diagnosis of acute gangrenous cholecystitis. AB - BACKGROUND: Factors previously identified by multivariate logistic regression that were predictive for gangrenous cholecystitis (GC) were used to develop a predictive equation. Our objective was to evaluate the sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values of this equation for detecting GC in patients with acute cholecystitis (AC). METHODS: Medical records of patients who presented to a tertiary care hospital with AC were reviewed. Twenty-one patient and clinical variables were recorded. We prospectively tested the results of the following equation against pathologic diagnosis: P=e((0.7116+0.9944.DM+1.7157.WBC-1.0319.ALT.2.0518.ALP+2.7078.PCF))/(1+e([ 0.7116+0.9944.DM+1.7157.WBC-1.0319.ALT-2.0518.ALP+2.7078.PCF])), where P = predicted value; DM = diabetes mellitus; WBC = white blood cell count; ALT = alanine aminotransferase; AST = aspartate aminotransferase; and PCF = pericholecystic fluid. RESULTS: Ninety-eight patients presented with AC and 18% had GC (18 of 98). Using a cutoff of P = 0.724, our equation had a specificity of 93%, sensitivity of 83%, PPV of 71%, and NPV of 96%, P <0.001 for the detection of GC. CONCLUSIONS: Our study demonstrates the equation may be useful in detecting the subset of AC patients who have GC. PMID- 15546552 TI - Extracellular pressure stimulates colon cancer cell adhesion in vitro and to surgical wounds by Src (sarcoma protein) activation. AB - BACKGROUND: We hypothesized that pressure stimulates colon cancer cell adhesion to surgical wounds. METHODS: We quantitated adhesion of murine 26/51 transplantable colon cancer cells by cell counting or chromium 51-labeling. Tumor cells were added to murine surgical wounds after 30 minutes preincubation under ambient or 15 mm Hg increased pressure. Src activation was assayed by immunoblotting for phosphorylated Src and inhibited by 4-amino-5-(4chlorophenyl) 7-(t-butyl)pyrazolo-[3-4-d]pyrimidine (PP2). RESULTS: Pressure stimulated colon 26/51 cell adhesion to murine wounds by 43% to 52% (n = 9, P <0.05 each). Pressure stimulated Src phosphorylation by 39% +/- 4% (n = 5, P = 0.004) in colon 26 cells. The Src inhibitor PP2 (20 mumol/L) did not inhibit Src phosphorylation at ambient pressure but prevented pressure stimulation of Src phosphorylation. Src blockade by PP2 did not affect basal adhesion of either tumor to murine wounds but completely blocked pressure stimulation of adhesion (n = 4, P <0.001 each). CONCLUSIONS: Increased pressure may activate cancer adhesion to surgical wounds via Src. Src antagonists might inhibit this process. PMID- 15546553 TI - Intraoperative regional myocardial acidosis predicts the need for inotropic support in cardiac surgery. AB - OBJECTIVE: To determine the impact of regional myocardial acidosis encountered during cardiac surgery on the need for inotropic and intra-aortic balloon (IAB) support. METHODS: Intramyocardial tissue pH(37C) was measured in 247 patients undergoing cardiopulmonary bypass (CPB). Inotropic support (INO) was defined as requiring one or more of norepinephrine/epinephrine/amrinone/dobutamine/>2.5 mug/kg/min dopamine, for at least 45 minutes intraoperatively, and intraoperative or postoperative IAB use. PH (corrected to 37 degrees C, pH(37C)) during surgery was compared in patients who needed INO versus those who did not. Multivariate logistic regression models identified the determinants of INO. RESULTS: Fifty patients (20.2%) required INO intraoperatively. pH(37C) was significantly lower throughout reperfusion in patients needing INO. Preoperative ejection fraction and pH(37C) during reperfusion were identified as independent predictors of INO. CONCLUSIONS: This is the first study to show that intraoperative regional myocardial acidosis, a preventable condition, independently determines the need for intraoperative INO. Increased INO is associated with greater postoperative mortality and morbidity. PMID- 15546554 TI - The impact of a novel resident leadership training curriculum. AB - BACKGROUND: Today's complex health care environment coupled with the 80-hour workweek mandate has required that surgical resident team interactions evolve from a military command-and-control style to a collaborative leadership style. METHODS: A novel educational curriculum was implemented with objectives of training the residents to have the capacity/ability to create and manage powerful teams through alignment, communication, and integrity integral tools to practicing a collaborative leadership style while working 80 hours per week. Specific strategies were as follows: (1) to focus on quality of patient care and service while receiving a high education-to-service ratio, and (2) to maximize efficiency through time management. RESULTS: This article shows that leadership training as part of a resident curriculum can significantly increase a resident's view of leadership in the areas of alignment, communication, and integrity; tools previously shown in business models to be vital for effective and efficient teams. CONCLUSION: This curriculum, over the course of the surgical residency, can provide residents with the necessary tools to deliver efficient quality of care while working within the 80-hour workweek mandate in a more collaborative style environment. PMID- 15546555 TI - Rebleeding and survival after acute lower gastrointestinal bleeding. AB - BACKGROUND: Previous studies of acute lower gastrointestinal bleeding (LGIB) have focused on evaluation and therapy. Measurement of long-term outcome has been rare. The purpose of this study was to document rebleeding and survival rates in patients with acute LGIB. METHODS: A retrospective review of all patients undergoing technetium-labeled red blood cell scans for LGIB from January of 1997 to December of 2002 was performed. Rebleeding was defined as identification of enteric bleeding requiring a transfusion 2 or more weeks after the initial bleeding episode. RESULTS: A total of 119 patients met inclusion criteria. Rebleeding was documented in 14 of 102 patients surviving for more than 2 weeks. The actuarial rebleeding rate was 15% at 2 years. No factors were identified that portended a higher likelihood of rebleeding. The 30-day mortality was 18% and the median survival was 60 months for the entire cohort. Of the 36 patients in whom cause of death was documented, 4 died of surgical complications and a single patient died as a direct result of hemorrhage. CONCLUSIONS: Rebleeding after an initial episode of LGIB occurs in a small percentage of individuals. Although survival is poor for patients with LGIB, few patients die as a direct consequence of hemorrhage. PMID- 15546556 TI - Effects of surgical gloves on postoperative peritoneal adhesions and cytokine expression in a rat model. AB - BACKGROUND: Postoperative intraperitoneal adhesions are a major cause of morbidity. We studied the effects of synthetic and latex gloves, and their powders, on postoperative adhesions and cytokine expression in a rat model. METHODS: Rats underwent laparotomy and cecal abrasion. Rats were grouped based on the glove type used: synthetic powder-free (SPF), synthetic powdered (SP), latex powder-free (LPF), and latex powdered (LP). Serum cytokine (tumor necrosis factor [TNF], interleukin-1 [IL-1], and IL-6) levels were measured. Animals were killed and peritoneal adhesions were graded. RESULTS: The SPF group had no adhesions. Adhesions were increased similarly in the SP and LPF groups, and further increased in the LP group. Postoperative serum cytokine levels showed a similar pattern of increases. CONCLUSIONS: The presence of latex or powder on surgical gloves promoted increased adhesions. Serum cytokine levels correlated with the degree of adhesion formation. Strategies to use latex-free, powder-free gloves and/or limit cytokine expression may decrease peritoneal adhesions in the clinical setting. PMID- 15546557 TI - Thirty-day and one-year predictors of death in noncardiac major surgical procedures. AB - BACKGROUND: We evaluated the predictive value of the American College of Cardiology/American Heart Association (ACC/AHA) cardiac risk classification, as well as other potential risk factors (procedure risk, smoking, obesity, hyperlipidemia, and renal insufficiency), on all-cause mortality at 30 days and at 1 year postoperatively. METHODS: In the year 2000, 1238 consecutive patients undergoing general anesthesia for various noncardiac surgical procedures at the Houston Veterans Affairs Medical Center were screened preoperatively and classified according to the ACC/AHA guidelines. Patients' charts were reviewed for the above-mentioned risk factors. RESULTS: A logistic regression analysis demonstrated that older age and higher procedure risk were associated with higher 30-day mortalities (P = 0.0012 and 0.0441, respectively). The ACC/AHA classification was positively correlated with mortality at 1 year (P = 0.0071). CONCLUSIONS: The ACC/AHA classification predicts mortality at 1 year but not at 30 days for major noncardiac surgeries; procedure-related risk is a better predictor of 30-day postoperative mortality in our patient population. PMID- 15546558 TI - Incidence and predictors of appendiceal tumors in elderly males presenting with signs and symptoms of acute appendicitis. AB - BACKGROUND: Patients with appendiceal tumors (AT) frequently present with the same signs and symptoms as acute appendicitis (AA). The purpose of this study was to identify the incidence of AT in a population of patients greater than 60 years of age thought to have AA, and to identify presenting factors that can help differentiate the 2 disease processes. METHODS: An institutional review board approved, retrospective review was performed identifying all patients greater than 60 years old who underwent either appendectomy or colectomy after presenting with signs and symptoms of AA from Janaury 1997 to April 2004. Patient records were examined for demographic variables, presenting signs and symptoms, and pathology. RESULTS: A total of 34 patients fit the entry criteria; 8 (24%) had AT. Average duration of symptoms was significantly longer (4.9 +/- 1.9 versus 2.3 +/- 0.3 days; P = 0.01) and hematocrit lower (36.5 +/- 2.3 versus 42.4 +/- 1.0%; P = 0.01) for patients with AT compared with patients with AP. No other significant differences in presenting signs and symptoms were noted. CONCLUSIONS: There is an increased incidence of appendiceal neoplasms among elderly patients presenting with signs and symptoms of AA. Distinction between AT and appendicitis is difficult preoperatively, but AT should be suspected when patients present with longer duration of symptoms or reduced hematocrit. PMID- 15546559 TI - Synergistic effects of Cox-1 and -2 inhibition on bladder and prostate cancer in vitro. AB - BACKGROUND: We hypothesized that combined treatment with cyclooxygenase (Cox)-1 (catechin) and Cox-2 (NS398)-specific inhibitors would reduce cellular proliferation synergistically in genitourinary cancer. METHODS: Bladder (T24 and TCCSUP) and prostate (DU145, LnCaP, and PC3) cancer cell lines were treated with catechin and NS398 at a dose of 100 mumol/L as single and combined treatments. Viability was measured by MTT assay at 24 and 72 hours. RESULTS: Significant synergism of Cox-1 and Cox-2 inhibitors was observed in both bladder cancer lines at both 24 and 72 hours. Synergism of Cox-1 and -2 inhibitors also was noted in the DU145 cells at 72 hours, LnCap cells at 24 hours, and PC3 at both 24 and 72 hours. CONCLUSIONS: Significant synergistic effects exhibited by the combination of Cox-1 and Cox-2 inhibitors suggest that these could become a highly effective treatment modality for carcinoma of both the bladder and prostate. PMID- 15546560 TI - Bile-pancreatic juice exclusion increases cholinergic M3 and CCK-A receptor expression and interleukin-6 production in ligation-induced acute pancreatitis. AB - BACKGROUND: Using an original model, the Donor Rat Model, we showed that bile pancreatic juice (BPJ) exclusion from gut exacerbates ligation-induced acute pancreatitis in rats. We also showed that muscarinic cholinergic M3 and CCK-A receptor expression is induced following duct ligation. Increased receptor number potentially could exacerbate cytokine production. We hypothesize that BPJ exclusion is responsible for M3 and CCK-A receptor induction and increased interleukin-6 (IL-6) production. METHODS: M3 and CCK-A receptor expression and IL 6 production were compared in rat pancreata 1 to 3 hours after duct ligation with or without BPJ replacement. RESULTS: Our studies showed that BPJ replacement attenuates duct ligation-induced increases in M3 and CCK-A receptor expression and IL-6 production. CONCLUSIONS: In this model, BPJ exclusion from gut induces M3 and CCK-A receptor expression and increases IL-6 production. In this experimental corollary of gallstone pancreatitis, BPJ exclusion from gut may play a key role in the mechanism of disease pathogenesis. PMID- 15546561 TI - Peptide YY inhibits the growth of Barrett's esophageal adenocarcinoma in vitro. AB - BACKGROUND: Peptide YY (PYY) is an endogenous gut hormone that inhibits the growth of certain cancers. Adenocarcinoma of the esophagus usually arises from Barrett's esophagus. We hypothesized that treatment of Barrett's adenocarcinoma with PYY would result in decreased proliferation. METHODS: Barrett's cancer cell lines (BIC and SEG-1) were treated with PYY (3-36) at 500 pmol/mL. Viability was measured by MTT at 24 and 72 hours. Apoptosis and necrosis was evaluated by flow cytometry. RESULTS: PYY reduced proliferation in SEG-1 cells at 24 hours (21.2% +/- 3.4%, P <0.001) and 72 hours (14.2% +/- 6.2%, P <0.001). In the BIC cells, growth was inhibited by 7.9% +/- 7.0%, P = 0.021 after 72 hours. PYY increased late apoptotic activity in SEG-1 cells by 31%, P = 0.014. CONCLUSIONS: This is the first report of antiproliferative effects of PYY against Barrett's carcinoma in vitro. Reductions in cell growth appear to be mediated by proapoptotic mechanisms. Further investigation of PYY in the treatment of Barrett's adenocarcinoma is warranted. PMID- 15546562 TI - Does the National Board of Medical Examiners' Surgery Subtest level the playing field? AB - BACKGROUND: We sought to determine if scores on the National Board of Medical Examiners' Surgery Subject Examination (NBME-SS) correlated with ward assessment of Fund of Knowledge and how the NBME-SS scores impacted surgery clerkship grade. METHODS: Student evaluations from 4 academic years (1999-2003) were analyzed. Scores from the Fund of Knowledge subsection were compared with scores on the NBME-SS. Student final grades were calculated with (at a weight of 10%) and without NBME-SS score. RESULTS: Data were available for all 399 students who rotated on surgery during the study period. Fund of Knowledge score and NBME-SS scores correlated minimally (Pearson's r = 0.108, P = 0.031). Performance on the NBME-SS helped 11% of students' grades, but 64% of students' grades were hurt. Fifteen percent of the students missed receiving honors because of their performance on the NBME-SS. CONCLUSIONS: The NBME-SS negatively influenced the grades of a large proportion of students and correlated only minimally with Fund of Knowledge scores as assessed on ward evaluations. PMID- 15546563 TI - Defining the learning curve for laparoscopic splenectomy for immune thrombocytopenia purpura. AB - BACKGROUND: The current study was undertaken to define the learning curve for laparoscopic splenectomy (LS) in patients with immune thrombocytopenic purpura (ITP). METHODS: The data of 50 patients who underwent LS for ITP between March 1996 and February 2003 were reviewed. Patients were divided into sequential groups of 10. Operative time, estimated blood loss, conversion to open procedure, length of stay (LOS), time to oral intake, complications, and mortality rates were analyzed. RESULTS: The mean OR time in the 3rd, 4th, and 5th groups of 10 were significantly shorter than the 1st and 2nd groups of 10. There were no significant differences in estimated blood loss, LOS, or time to oral intake between the groups. Three conversions to open splenectomy occurred; one each in the 2nd, 3rd, and 4th groups of 10. Complications were evenly distributed between groups. There were no deaths. CONCLUSION: The learning curve for LS in patients with ITP is a minimum of 20 cases. PMID- 15546564 TI - Microvascular responses to adenosine help explain functional and pathologic differences between intestinal segments. AB - BACKGROUND: Many physiologic (post-prandial hyperemia), pathologic (inflammatory bowel disease), and clinical (enteral feeding) phenomena involve changes in microvascular blood flow to the intestine. Adenosine (Ado) derived from energy metabolism causes vasodilation and appears to be involved in some of these events. The Ado-mediated control mechanisms appear to vary with the diameter of the microvessels and the function of the tissue. This suggests the possibility that Ado-based microvascular control varies between anatomic intestinal segments and microvascular levels in those intestinal segments. METHODS: In vivo digital intravital microscopy was used to measure the responses of larger distributing (A1) and smaller premucosal (A3) vessels to Ado in intact neurovascular loops of jejunum and terminal ileum of the rat. Dose-response curves to Ado were determined. RESULTS: Microvascular dilation and augmented blood flow to Ado were significantly greater in the jejunum than in the terminal ileum. Ado-induced dilation was greater in the smaller A3 than in the larger A1 microvessels. DISCUSSION: These data indicate (1) different vasodilator mechanisms for the jejunum and the terminal ileum, (2) a greater role for Ado-related microvascular control in the jejunum compared with the ileum, and (3) a greater Ado-related control in the premucosal (A3) vessels. These findings suggest that Ado-mediated microvascular effects could explain why some clinical phenomena vary in intensity in selective intestinal segments, and are likely to involve different microvascular control mechanisms in the different segments. Knowledge of these Ado mechanisms could be beneficial in certain clinical scenarios to control blood flow during pathologic conditions. PMID- 15546565 TI - Temozolomide is a novel regional infusion agent for the treatment of advanced extremity melanoma. AB - BACKGROUND: Regional infusion therapy with melphalan (LPAM) is an accepted treatment for advanced extremity melanoma. However, much room exists for improving the therapeutic index of this type of therapy. METHODS: Isolated limb infusion (ILI) with temozolomide (TMZ), a novel methylating agent, was performed using a nude rat bearing human melanoma xenograft. Additional rats were treated systemically with TMZ, or regionally with LPAM or 10% dimethyl sulfoxide (DMSO; control) using ILI. RESULTS: Rats that received systemic TMZ showed a poor tumor response and no tumor regression. In contrast, intra-arterial TMZ demonstrated a prolongation of tumor growth delay in a dose-responsive manner. In comparison with LPAM of equitoxic dose, TMZ provided both longer tumor growth delay and a greater number of tumor regressions. CONCLUSIONS: These data suggest that ILI with TMZ is an effective treatment for advanced extremity melanoma and may be better than LPAM in this setting. PMID- 15546566 TI - Characterization of endoleaks by dynamic computed tomographic angiography. AB - BACKGROUND: Current imaging modalities may not be able to detect endoleaks, differentiate between type II and type III, or localize inflow and outflow sources. We describe a new technique that can characterize endoleaks to guide secondary intervention. METHODS: One hundred four patients with Zenith (Cook, Inc.) endograft repair of abdominal aortic aneurysms (AAAs) were monitored by serial computed tomographic angiography (CTA). Endoleaks were evaluated with a dynamic CTA using a stationary table position, 24-mm beam collimation, and continuous scanning over 30 to 40 seconds to create a cine. RESULTS: Twelve patients (12%) had endoleaks that persisted or appeared more than 30 days post deployment. Five patients in whom the standard CT surveillance protocol could not differentiate type II versus type III endoleaks underwent dynamic CTA. This technique accurately characterized the endoleaks and localized inflow and outflow branches to guide the subsequent successful secondary interventions. CONCLUSIONS: Dynamic CTA is a useful technique to evaluate endoleaks for characterization and precise localization to guide secondary interventional therapy. PMID- 15546567 TI - Short-stay carotid endarterectomy in a tertiary-care Veterans Administration hospital. AB - BACKGROUND: This study focused on 200 carotid endarterectomies (CEA) performed at our Veterans Administration Hospital (VAH) to determine whether 1-day hospitalization after CEA is safe and the degree to which it can be achieved. METHODS: Over 36 months, 200 CEAs were performed for asymptomatic stenosis (n = 104), transient ischemic attacks (n = 68), and stroke (n = 28). General anesthesia was used in 189 procedures. RESULTS: The hospital stay was 1 day for 132 procedures and more than 1 day in 68 CEAs. The average stay was 1.69 +/- 1.5 days. After surgery there were 3 strokes, 5 hematomas that required evacuation, and 5 myocardial infarctions. There were no deaths. Four patients were readmitted in the 1-day and the greater than 1-day stay groups. History of myocardial infarction, renal insufficiency, longer operative time, and complications correlated with a greater than 1-day stay (P <0.05). CONCLUSION: A 1-day hospital stay is safe and practical in a VAH setting, resulting in good clinical outcomes. PMID- 15546568 TI - Measurement of laparoscopic skills in microgravity anticipates the space surgeon. AB - BACKGROUND: There clearly must be a surgical capability to support the exploration of the solar system by humans. We hypothesized that microgravity would impair surgical task performance and sought to measure degree and criticality of task erosion. METHODS: Twenty subjects performed basic laparoscopic exercises (grasping, cutting, clipping, and suturing) in an inanimate trainer on the ground and during the microgravity phase of a parabolic flight. Skill assessment involved time, accuracy, and force transfer. RESULTS: A decrease in the total number of on-target task completions was observed in microgravity compared with the ground performance for all tasks. The ratio between the total number of attempts and the number of completions was higher in microgravity. The amount of force introduced into the system was not significantly higher in microgravity. CONCLUSIONS: We conclude that although tasks are performed with much greater deliberation and work in microgravity, there is no barrier to accurate and effective laparoscopic surgery. PMID- 15546569 TI - Complications and long-term survival for alcoholic patients with resectable lung cancer. AB - BACKGROUND: The aim of this study was to determine the surgical risks and long term survival in alcoholic patients undergoing resection for non-small-cell lung cancer. METHODS: Nineteen resected patients comprising the alcoholic group were identified by either a Diagnostic and Statistical Manual of Mental Disorders-IV diagnosis of alcohol dependence/abuse, or an alcohol consumption of 60 oz/d or more. Alcoholic patients were compared with 37 nonalcoholic patients undergoing resection. RESULTS: Alcoholic patients had an increase in major infectious complications (37% [7 of 19] versus 5% [2 of 37], P = 0.005), respiratory failure (42% [8 of 19] versus 5% [2 of 37], P 30 days) developed in 8 (20%) patients. Percent excess weight loss at 3, 6, and 12 months was 44% (n = 34), 59% (n = 29), and 70.0% (n = 22), respectively. In 23 patients who were followed-up for more than 3 months, DM resolved in 79% and improved in 21% at a mean follow up evaluation of 13 months. CONCLUSIONS: Laparoscopic RYGB can be performed with acceptable morbidity and with good short-term results in a VA hospital setting. Morbid obesity is prevalent in the VA patient population and access to bariatric surgery should be an available alternative. PMID- 15546582 TI - Improvements in dialysis access survival with increasing use of arteriovenous fistulas in a Veterans Administration medical center. AB - BACKGROUND: Native arteriovenous fistulas (AVFs) have been found to exhibit higher survival rates and lower complication rates than prosthetic grafts (AVGs). METHODS: Between August 2001 and December 2003, 93 patients with end stage renal disease (ESRD) had primary dialysis access placed at a single Veterans Administration medical center. Of these 93 patients, 67 had AVFs created and 26 patients had AVGs implanted. RESULTS: The percentage of patients who did not require additional intervention was 84% (56 of 67) for AVF and 78% (21 of 26) for AVG after 4 to 31 months of follow-up evaluation. In the AVF group, repeat interventions were as follows: collateral ligation (4), angioplasty owing to central stenosis (2), AVF ligation due to arterial steal phenomenon (1), and new AVF creation owing to clotting (1). Four AVFs were later converted to AVG. In the AVG group there were 4 venous anastomosis stenosis seen in 3 patients who required angioplasty. Two patients needed thrombectomy and revision, and 1 graft was removed because of infection. AVF prevalence in our dialysis patients was 63%, with 33% AVG and 4% temporary catheter. CONCLUSIONS: The National Kidney Foundation-Dialysis Outcome Quality Initiative (NKF-DOQI) guidelines for dialysis access reawakened interest in maximizing the use of renal veins for AVF. AVFs created by using the patient's native vein provides the best vascular access for dialysis when compared with prosthetic grafts. AVF has better long-term patency with fewer complications. PMID- 15546583 TI - The spectrum of colovesical fistula and diagnostic paradigm. AB - BACKGROUND: Our experience with colovesical fistula (CVF) over a 12-year period was reviewed to clarify its clinical presentation and diagnostic confirmation. METHODS: Twelve patients with CVF were identified. Presenting symptoms, etiologic factors, diagnostic investigations, and subsequent treatment were reviewed. RESULTS: Underlying etiologies were diverticular disease (75%), colon cancer (16%), and bladder cancer (8%). Pneumaturia (77%) was the most common presentation, followed by urinary tract infections, dysuria and frequency (45%), fecaluria (36%), hematuria (22%), and orchitis (10%). The ability of various preoperative investigations to identify a CVF were: computed tomography (CT) (90%), barium enema (BE) (20%), and cystography (11%), whereas cystoscopy, intravenous pyelogram (IVP), and colonoscopy were nondiagnostic. All patients underwent single- or multiple-staged repair of the fistula. CONCLUSIONS: In patients with a suspected CVF, we recommend CT followed by a colonoscopy as a first-line investigation to rule out malignancy as a cause of CVF. Other modalities should only be used if the diagnosis is in doubt or additional information is needed to plan operative management. PMID- 15546584 TI - Jugular vein aneurysm or phlebectasia. PMID- 15546585 TI - Platelet glycoprotein Ia C807T, Ib C3550T, and IIIa Pl(A1/A2) polymorphisms and ischemic stroke in young Taiwanese. AB - Platelet plays a pivotal role in the pathogenesis of thrombotic cardiovascular diseases. Recently, the polymorphism of platelet glycoprotein (GP) genes has been reported to be associated with an increased risk for ischemic stroke. The purpose of this study is to evaluate the association between platelet GP genetic variants and ischemic stroke in young Taiwanese. We conducted a case-control study in 157 young ischemic stroke patients recruited between September 2001 and March 2003 and 157 age- and sex-matched controls. The genotypes of platelet GP Ia C807T, GP Ib C3550T, and GP IIIa Pl(A1/A2) polymorphisms were analyzed by polymerase chain reaction-restriction fragment length polymorphism. Student's t-test, chi-square test, and logistic regression modeling were used for data analyses. The GP Ia C807T CC, CT and TT genotype frequencies were similar between patients (50.3%, 43.9%, 5.7%) and controls (53.5%, 38.9%, 7.6%; p=0.58). There were no significant differences in GP Ib C3550T CC and CT genotype distributions between patients (91.1%, 8.9%) and controls (91.7%, 8.3%; p=0.84). Of all subjects, none carries GP IIIa Pl(A2) mutation. In conclusion, platelet GP Ia C807T and GP Ib C3550T polymorphisms in our population are less common compared with Caucasians, and GP IIIa Pl(A1/A2) genetic mutation is not found, and all of them are not associated with ischemic stroke in young Taiwanese. PMID- 15546586 TI - Characterization of cellular and neurological damage following unilateral hypoxia/ischemia. AB - Rodent models of stroke are often used to investigate the mechanisms that lead to ischemic neuronal damage. In this study, we used a model of cerebral hypoxia with ischemia to produce unilateral damage in C57Bl/6 mice. Lesion volume, ascertained by TTC staining, increased with longer durations of hypoxia. Additionally, cresyl violet, TUNEL, and FluoroJade staining showed a statistically significant increase in cellular damage in the ipsilateral cortex, CA1 pyramidal layer, and dentate gyrus of the hippocampus of ipsilateral hypoxic/ischemic tissue versus sham tissue. Astrocyte reactivity, determined by GFAP staining, was significantly higher in the ipsilateral H/I cortex and contralateral hippocampus compared to sham cortex and hippocampus, respectively. Increased microglia activation was evident in the H/I-treated cortex and hippocampus versus sham cortex and hippocampus, particularly within areas undergoing degeneration. To examine whether this model produces motor deficits, a battery of tests were administered before and after hypoxia. Following 45 min H/I, locomotor activity, rotarod performance and performance on an inverted wire hang test were all significantly decreased. These data indicate that the histological evidence of neuronal damage is consistent with functional deficits and suggest that this model may be useful for investigating strategies designed to protect neurons from hypoxia/ischemia induced damage. PMID- 15546587 TI - HLA typing in focal myositis. AB - It is still controversial if idiopathic focal myositis is a part of systemic polymyositis. We present here four patients, including identical twins, with focal myositis accompanied by the same HLA typings. Gradually developing unilateral calf muscle pain was an initial symptom in all patients. Neither muscular weakness nor creatine kinase (CK) elevation was observed, while minimal inflammatory findings such as erythrocyte sedimentation rate (ESR) increase appeared in serum. Magnetic resonance imaging (MRI) revealed localized abnormalities of calf muscles. Biopsy specimen was characterized by perimysial and endomysial inflammatory infiltration consisted of T cells and macrophages and rare necrotic fibers. Corticosteroid administrations ameliorated their symptoms and signs, though recurrence occurred along with decreasing doses. HLA typings common to all patients were A2, B62, Cw3, and DQ3, whereas HLA-D DNA typings were DQB1 *0303 for two patients, and DQB1*0302 for three patients. These findings suggest that at least some focal myositis may be a new disease unit, with a common genetic background but not a part of systemic polymyositis. PMID- 15546588 TI - Increased incidence of the Hfe mutation in amyotrophic lateral sclerosis and related cellular consequences. AB - The etiology of amyotrophic lateral sclerosis (ALS) is unknown. The presence of mutations in the superoxide dismutase gene (SOD1) has led to theories regarding a role for oxidative stress in the pathogenesis of this disease. A primary cause of oxidative stress is perturbations in cellular iron homeostasis. Cellular iron mismanagement and oxidative stress are associated with a number of neurodegenerative diseases. One mechanism by which cells fail to properly regulate their iron status is through a mutation in the Hfe gene. Mutations in the Hfe gene are associated with the iron overload disease, hemochromatosis. In the current study, 31% of patients with sporadic ALS carried a mutation in the Hfe gene, compared to only 14% of patients without identifiable neuromuscular disease, or with neuromuscular diseases other than ALS (p<0.005). To determine the cellular consequences of carrying an Hfe mutation, a human neuronal cell line was transfected with genes carrying the Hfe mutation. The presence of the Hfe mutation disrupted expression of tubulin and actin at the protein levels potentially consistent with the disruption of axonal transport seen in ALS and was also associated with a decrease in CuZnSOD1 expression. These data provide compelling evidence for a role for the Hfe mutation in etiopathogenesis of ALS and warrant further investigation. PMID- 15546589 TI - Hereditary sensory neuropathy type 1 in a Portuguese family-electrodiagnostic and autonomic nervous system studies. AB - Hereditary sensory and autonomic neuropathy type 1 (HSAN 1) is a dominantly inherited disorder; its gene locus is mapped on chromosome 9q22. Three different missense mutations (C133Y, C133W and V144D) have been described in 11 families from Australia, England and Austria. Common clinical features have been found in these families. We report the clinical and electrophysiological features of three members of a large Portuguese family with HSAN 1 and the C133Y missense mutation. The affected members showed typical clinical features. Electrophysiological findings were consistent with a distal axonal predominantly sensory neuropathy with motor involvement, in three different severity stages. No autonomic involvement was detected in sudomotor and cardiovascular tests. This report documents the lesion of the motor nerve fibers in this disease, as well as the preservation of the autonomic nervous system function, therefore suggesting that HSNA is an inappropriate name for this disorder. PMID- 15546590 TI - Effect of immunotherapy in myelitis with atopic diathesis. AB - OBJECTIVE: A recent nationwide survey of myelitis with atopic diathesis in Japan disclosed that the disease frequently shows a chronic persistent course. A neuropathological study of the spinal cord also revealed chronic active inflammation. Since the effects of various immunotherapies have not been studied extensively in this condition, we evaluated the efficacies of various immunotherapies in patients with myelitis with atopic diathesis. PATIENTS AND METHODS: Forty-two treatments in 26 patients with myelitis with atopic diathesis were retrospectively analyzed. One of the following therapies was administered: (1) corticosteroids (CS) (pulse therapy followed by oral administration with gradual tapering); (2) intravenous immunoglobulin (IVIG) (400 mg/kg/day for 5 consecutive days); (3) plasma exchanges (PE); or (4) PE followed by IVIG or CS (PE+IVIG/CS). The therapeutic efficacies were evaluated by thorough neurological examination and laboratory tests including MRI, somatosensory evoked potentials (SEPs) and motor evoked potentials (MEPs). RESULTS: Objective neurological findings improved in 89% of the PE group and 90% of the PE+IVIG/CS group, compared with only 72% of the CS and 60% of the IVIG groups. Improvement determined by laboratory tests was seen in 57% of the PE and 57% of the PE+IVIG/CS groups, compared with only 15% of the CS and none of the IVIG groups. Thus, the improvement rate determined by laboratory tests was significantly greater for therapies including PE than for those without PE (p=0.0187). CONCLUSIONS: These data suggest that immunotherapy is effective in myelitis with atopic diathesis despite a chronic persistent course, and that PE is the most beneficial immunotherapy. PMID- 15546591 TI - Release of neurobiochemical markers of brain damage is related to the neurovascular status on admission and the site of arterial occlusion in acute ischemic stroke. AB - OBJECTIVES: The study aimed at an analysis of the kinetics of protein S100B and neuron-specific enolase (NSE) and their relation to the site of arterial occlusion in patients with acute ischemic stroke. METHODS: We investigated 32 consecutive patients admitted within 6 h after stroke onset. Serial venous blood samples were taken hourly between 1 and 6 h, and at 12, 18, 24, 48, 72, 96, and 120 h after stroke onset. The neurovascular status was assessed on admission and monitored by repetitive extracranial and transcranial duplex sonography. In all patients, infarct volume was calculated. The neurological deficit was quantified by the National Institutes of Health stroke scale score, and functional outcome after 3 months was assessed with the modified Rankin Scale. RESULTS: Patients with normal flow velocities in basal cerebral arteries at admission showed significantly less S100B release than those with main stem or multiple branch occlusions (p<0.01). S100B cut-off values of 0.15 microg/l (between 6 and 18 h), 0.21 microg/l (between 24 and 48 h) and 0.5 microg/l (from 72 to 120 h) differentiated best between patients with initially normal and pathological sonographic vessel findings. The release of S100B and NSE was highly correlated with the severity of the corresponding neurological deficit as well as with the final infarct volume. S100B concentrations from 6 h on were associated with the functional outcome. S100B values 48 h after stroke above 0.2 microg/l indicated a poor functional status 3 months after stroke. CONCLUSIONS: Protein S100B may serve as a monitoring parameter in acute ischemic stroke, especially with respect to the neurovascular status. Furthermore, S100B obtains additional information about functional outcome. PMID- 15546592 TI - The clinical spectrum of musical hallucinations. AB - Musical hallucinations are a well known although rare phenomenon in neurological and psychiatric patients. Many case reports have been published to date. However, an accepted common theory on the classification and on the pathophysiology of musical hallucinations is still missing. We analysed all cases published to date, including two own cases, with respect to their demographic and clinical features and to the possible pathomechanisms underlying the hallucinations. In total, 132 cases could be analysed statistically and separated into five groups according to their aetiology (hypacusis; psychiatric disorder; focal brain lesion; epilepsy; intoxication). There was a female preponderance of 70% and a mean age of 61.5 years. Patients with focal brain lesions were significantly younger than the other groups, the hemisphere of the lesion did not play a major role. No systematic studies on treatment are available. However, anticonvulsant and antidepressive substances were reported to be effective most consistently. The pathophysiology of musical hallucinations is discussed considering the theories of deafferentiation including the concept of auditory Charles-Bonnet syndrome, of sensory auditory deprivation, of parasitic memory, and of spontaneous activity in a cognitive network module. In conclusion, musical hallucinations are a phenomenon with heterogeneous clinical and pathophysiological backgrounds. PMID- 15546593 TI - rTMS of the prefrontal cortex in the treatment of chronic migraine: a pilot study. AB - A recent fMRI study showed that dorsolateral prefrontal cortex (DLPFC) exerts an inhibitory control on pain pathways in humans. We investigated whether high frequency rTMS over left DLPFC could ameliorate chronic migraine. Treatment consisted of 12 rTMS sessions, delivered in alternate days over left DLPFC. Sham rTMS was used as placebo. Eleven patients were randomly assigned to the rTMS (n=6) or to the placebo (n=5) treatment. Measures of attack frequency, headache index, number of abortive medications (outcome measures) were recorded in the month before, during and in the month after treatment. Subjects treated by rTMS showed a significant reduction of the outcome measures during and in the month after the treatment as compared to the month before treatment. No significant differences in the outcome measures were observed in the placebo group. High frequency rTMS over left DLPFC was able to ameliorate chronic migraine. This is in agreement with the suggested role of DLPFC in pain control. PMID- 15546594 TI - Diffusion tensor MRI as a diagnostic tool of upper motor neuron involvement in amyotrophic lateral sclerosis. AB - BACKGROUND: Clinical identification of upper motor neuron (UMN) dysfunction in amyotrophic lateral sclerosis (ALS) is often difficult, particularly early in the course of the disease, or when lower motor neuron (LMN) dysfunction is prominent. Diffusion tensor MR imaging (DTI) can provide unique information on axonal organization by measuring diffusion anisotropy and the directionally independent diffusion. OBJECTIVE: The purpose of this study was to assess water diffusion changes along pyramidal tracts of the brainstem in patients with ALS and to investigate possible correlations between changes of diffusion properties and various clinical parameters. METHODS: We studied 16 patients (M:F=9:7, 50.5+/ 12.4 years) with ALS as defined by clinical and electrophysiological examinations. These patients were compared with 11 healthy, age and sex-matched controls (M:F=5:6, 54.5+/-9.9 years). DTI was performed using a single shot SE EPI with 25 noncollinear diffusion gradient directions (b=1000 s/mm(2)) and with no diffusion gradient on a 3.0-T MR system. RESULTS: By multifactorial ANOVA, the effects of group (patient versus control) and anatomical level on fractional anisotropy (FA) and mean diffusivity (MD) were significant (p<0.001 for both parameters), whereas the effect of side (left versus right) and interactions between factors (group by side and group by anatomical level) were not (p>0.05). In all subjects, FA and MD varied greatly depending on the anatomical level, and FA was highly variable even between contiguous slices in the pons and medulla, whereas relatively constant FA values were noted at the level of the midbrain. Cerebral peduncle was the only area that showed significant differences of diffusion properties between patients and controls (p<0.001 for FA, p=0.001 for MD). Correlation analysis revealed a significant inverse relationship between the FA value and the extent of UMN signs (r=-0.81, p<0.001). CONCLUSIONS: Alteration of diffusion properties in the cerebral peduncle in ALS may reflect pathological changes in structures rather than regional architectural variations of the corticospinal tracts or experimental artifacts. PMID- 15546595 TI - Chronological changes of sympathetic outflow to muscles in amyotrophic lateral sclerosis. AB - To confirm correlations between muscle sympathetic nerve activity (MSNA) and patients' chronological data, we selected 40 consecutive patients with sporadic amyotrophic lateral sclerosis (ALS) recorded by similar methods. MSNA at rest was quantified as the number of sympathetic bursts per 100 heartbeats and as the value expressed as a percentage of the predicted value based on control data. Twelve patients who underwent recordings of MSNA twice at intervals of 6 months or more showed marked decreases in MSNA amplitudes and frequencies between examinations. There was a slightly positive correlation between the frequency of MSNA and age, though younger patients exhibited higher values of MSNA than older patients. The standardized value of MSNA correlated negatively with disease duration and disability levels (p<0.01, 0.05, respectively), but several patients with duration shorter than 12 months showed low values of MSNA. Twelve patients who underwent repeated recordings of MSNA showed a significant decrease in the mean standardized value of MSNA (102.6+/-24.9%) at the second examination, compared to the value (114.3+/-18.9%) at the first one. In ALS, sympathetic outflow to muscles tends to increase initially and then decrease with increasing age and duration. This pattern may be similar to chronological changes at motor neurons. PMID- 15546596 TI - Primary angiitis of the central nervous system: report of five biopsy-confirmed cases from Colombia. AB - INTRODUCTION: Primary (isolated) angiitis of the central nervous system (PACNS) is a rare cause of cerebrovascular disease (CVD), and few leptomeningeal and brain biopsy (LBB)-confirmed cases have been reported from South America. METHODS: We retrospectively reviewed charts of patients with diagnosis of cerebral angiitis admitted between March 1991 and July 2001 to a single university hospital in Medellin, Colombia. Patients with definitive diagnosis of PACNS by Alrawi et al.'s LBB criteria were selected. We excluded other causes of cerebral angiitis as well as cases without LBB confirmation. RESULTS: We report five patients, four men and one woman, with a mean age at onset of 24.4 years, and an average disease progression of 12.4 days. Four presented with headache and motor weakness, three had seizures, and two had alterations of consciousness. Cerebral MRI was abnormal in all five cases; brain CT in four, and cerebral angiography in two. The cerebrospinal fluid (CSF) was abnormal in two patients. Leptomeningeal and brain biopsies revealed mononuclear infiltration in the wall of small blood vessels in all. Three had concurrent meningeal and cerebral involvement, two had necrotizing angiitis, and one had vascular and encephalitic lesions. All received only steroid treatment; the 1-year follow-up revealed good prognosis without relapses. CONCLUSION: We report five biopsy-proven cases of PACNS from Colombia associated with neurological and neuroimaging abnormalities; these patients presented a mild inflammatory disease that was correlated with few CSF abnormalities and good response to single steroid treatment without relapses. Leptomeningeal and brain biopsy is mandatory for a definitive diagnosis. PMID- 15546597 TI - Motor root conduction block in traumatic brachial plexopathy. AB - A 31-year old man sustained severe left brachial plexus traction injury. Electrical stimulation demonstrated multilevel motor root conduction block, which reversed after a 4-month period. Motor root conduction studies are useful diagnostic and prognostic adjuncts in the management of brachial plexopathy. PMID- 15546599 TI - Ischemia-reperfusion injury of peripheral nerve in experimental diabetic neuropathy. AB - The pathogenesis of human diabetic neuropathy likely involves the interplay of hyperglycemia, ischemia, and oxidative stress. Mild-moderate ischemia-reperfusion to streptozotocin (STZ)-induced diabetes results in florid fiber degeneration in diabetic but not in normal nerves. Uncertainty exists as to the influence of duration of diabetes on this susceptibility. We therefore studied diabetic tibial and sciatic nerves using a rat ischemia-reperfusion (IR) model after 1 month and 4 months of diabetes utilizing electrophysiological, behavioral, and neuropathological methods. Electrophysiological abnormalities were present in 1 month diabetic rats (D) and persisted over 4 months. Behavioral scores were decreased markedly at 4 months (p<0.05). Endoneurial edema and ischemia fiber degeneration (IFD) were observed at both the 1-month (p<0.01 and p<0.001) and 4 month (p<0.001) durations in diabetic nerves, whereas only mild or no damage was observed in age-matched control nerves. These findings demonstrate that STZ induced diabetes exacerbates the morphological and electrophysiological pathology in peripheral nerve to IR injury both in the early timepoint of 1 month and late timepoint of 4 months, although there was a gradation of injury, which is more severe at the later timepoint. Reperfusion exaggerated morphological pathology in 1-month STZ-induced diabetic peripheral nerve. PMID- 15546598 TI - Investigation of the low-density lipoprotein receptor gene and cholesterol as a risk factor for migraine. AB - The Low-Density Lipoprotein Receptor (LDLR) gene is a cell surface receptor that plays an important role in cholesterol homeostasis. We investigated the (TA)n polymorphism in exon 18 of the LDLR gene on chromosome 19p13.2 performing an association analysis in 244 typical migraine-affected patients, 151 suffering from migraine with aura (MA), 96 with migraine without aura (MO) and 244 unaffected controls. The populations consisted of Caucasians only, and controls were age- and sex-matched. The results showed no significant difference between groups for allele frequency distributions of the (TA)n polymorphism even after separation of the migraine-affected individuals into subgroups of MA and MO affected patients. This is in contradiction to Mochi et al. who found a positive association of this variant with MO. Our study discusses possible differences between the two studies and extends this research by investigating circulating cholesterol levels in a migraine-affected population. PMID- 15546600 TI - Plasma homocysteine, vitamin B12 and folate in Alzheimer's patients and healthy Arabs in Israel. AB - High plasma homocysteine (tHcy) is a risk factor for cardiovascular disease and stroke and Alzheimer's disease (AD). An inverse relationship has been reported between tHcy and plasma B12 and folate levels. Seventy-nine AD patients and 156 controls from three Arab villages in northern Israel participated. Plasma tHcy, B12 and folate levels were determined. Data were analyzed using univariate statistical tests and logistical regression with confounders. tHcy was significantly higher in AD patients (20.6+/-8.7 micromol/l) than in controls (16.4+/-6.5 micromol/l) (p=0.03) after correction for year of birth, gender and smoking status. Plasma B12 (322.9+/-136.0/350.5+/-175.3 pmol/l) and plasma folate (4.5+/-3.8/4.9+/-2.6 nmol/l) levels did not differ significantly between AD patients and controls. Subjects in the highest tHcy tertile or in the lowest B12 and folate tertiles did not have greater risk to develop AD. In this population residing in Arab villages in northern Israel, tHcy levels were significantly higher among AD patients than in controls. Plasma B12 and folate levels were lower among cases but were not significant. There was not a significant association between plasma tHcy, B12 and folate levels in controls or AD patients. High levels of tHcy may suggest the need for folate and vitamin B12 supplementation in this population. PMID- 15546601 TI - Fragile X gene premutation in multiple system atrophy. AB - Previous reports have suggested that expansion of the CGG repeat located in the fragile X mental retardation 1 (FMR1) gene might be responsible for a significant number of patients with the multiple system atrophy (MSA) phenotype. Analysis of 65 MSA patients found only 4.6% displayed CGG expansions in the suspected range. This is similar to the frequency reported in the normal population, suggesting that this expansion does not play a major role in the MSA phenotype. PMID- 15546602 TI - Small fiber neuropathy: a common and important clinical disorder. AB - Small fiber neuropathy (SFN) is a neuropathy selectively involving small diameter myelinated and unmyelinated nerve fibers. Interest in this disorder has considerably increased during the past few years. It is often idiopathic and typically presents with peripheral pain and/or symptoms of autonomic dysfunction. Diagnosis is made on the basis of the clinical features, normal nerve conduction studies (NCS) and abnormal specialized tests of small nerve fibers. Among others, these tests include assessment of epidermal nerve fiber density, temperature sensation tests for sensory fibers and sudomotor and cardiovagal testing (QSART) for autonomic fibers. Unless an underlying disease is identified, treatment is usually symptomatic and directed towards alleviation of neuropathic pain. PMID- 15546603 TI - Impairment in Guillain-Barre syndrome during the first 2 years after onset: a prospective study. AB - OBJECTIVES: To provide a comprehensive description of impairment in patients with Guillain-Barre syndrome (GBS) in Sweden during the first 2 years after disease onset. METHODS: In this prospective multi-centre study, 42 patients, mean age 52 years, were evaluated at 2 weeks, 2 months, 6 months, 1 year and 2 years. Evaluations made use of validated, reliable measures of muscle strength, grip strength, finger dexterity, balance, facial-muscle function, respiratory function, gait, motor performance and sensory examination, and included patients' own assessments of pain, fatigue and paraesthesia. RESULTS: Mechanical ventilation was required in 21% of patients. At 2 weeks, 1 year and 2 years after GBS onset: 100%, 62% and 55% of patients had submaximal overall muscle strength; 98%, 38% and 31% subnormal grip strength; and 38%, 14% and 12% affected facial muscle function. At the same time points, 62%, 10% and 7% of patients were unable to walk 10 m independently; and affected sensation was detected in 93%, 55% and 52%. CONCLUSIONS: Recovery occurred mainly during the first year after onset. At 2 years, motor impairment and sensory impairment were each still detectable in more than 50% of patients. We conclude that residual impairment is significant, somatically widespread and, likely, persistent. PMID- 15546604 TI - Central nervous system immunity associated with clinical outcome in acute encephalitis. AB - To search for useful laboratory measures of central nervous system (CNS) immunity that may provide an accurate prognosis or clues regarding treatment choice, cerebrospinal fluid (CSF) samples were obtained from 14 consecutive patients with acute encephalitis during acute as well as convalescent or chronic stages, and then examined for surface antigen expressions by lymphocytes and the presence of antineuronal tissue antibodies as well as the levels of IgG-related parameters and proinflammatory cytokines, including IL-2, IL-6, IFN-gamma, and tumor necrosis factor-alpha (TNF-alpha). Seven patients with aseptic viral meningitis served as nonencephalitic controls. Eight of the 14 acute encephalitis patients recovered fully, and reductions in the percentages of CD4(+)CD29(+) helper inducer T cells and IL-2 receptor-positive CD4(+) cells were associated with early recovery and favorable outcome, respectively, whereas a low percentage of CD4(+)CD26(+) memory T cells during an acute stage was associated with an unfavorable outcome following adjunctive intravenous corticosteroid treatment. Further, three of the four encephalitis patients who exhibited autoantibodies had a poor prognosis. These findings suggest that CNS immunity status has an effect on prognosis, while flow cytometric analyses of CSF CD4(+) helper T cell subsets may serve as effective means of assessment. PMID- 15546605 TI - Vitamins A, C and E in vascular parkinsonism. PMID- 15546607 TI - The genetic basis for the commitment to chronic versus acute infection in Pseudomonas aeruginosa. AB - The opportunistic pathogen Pseudomonas aeruginosa causes both acute and chronic airway infections. In a recent issue of Developmental Cell, Goodman et al. (2004) show that the RetS two-component gene regulatory module inversely controls expression of genes associated with acute and chronic infection. PMID- 15546608 TI - mRNA turnover meets RNA interference. AB - By using two very different approaches, recent work by Gazzani et al. (2004) and Souret et al. (2004) reveal a fundamental link between mRNA degradation and RNA silencing pathways in Arabidopsis. PMID- 15546609 TI - Good fences make good neighbors: barrier elements and genomic regulation. AB - In the genome, it is essential to maintain a physical barrier between active and inactive regions; however, the nature of this barrier has been elusive. In a recent issue of Molecular Cell, West et al. (2004) shed light on mechanisms underlying these molecular "fences." PMID- 15546610 TI - Telomeres thrown for a loop. AB - A remarkable paper from the de Lange lab (Wang et al., 2004) in a recent issue of Cell reveals that homologous recombination can result in the abrupt shortening of telomeres in a process that appears to involve reciprocal crossing over within the t-loop structure that protects chromosome ends. PMID- 15546611 TI - B cell development leads off with a base hit: dU:dG mismatches in class switching and hypermutation. AB - The mechanisms underlying somatic hypermutation (SHM) and class switch recombination (CSR) have been the subject of much debate. Recent studies from the Neuberger and Honjo labs have lent insight into these distinct processes, and we discuss a new, comprehensive model for how AID, uracil DNA glycosylase (UNG) and the mismatch repair system function in both SHM and CSR. PMID- 15546612 TI - Mastermind recruits CycC:CDK8 to phosphorylate the Notch ICD and coordinate activation with turnover. AB - Notch signaling releases the Notch receptor intracellular domain (ICD), which complexes with CBF1 and Mastermind (MAM) to activate responsive genes. We previously reported that MAM interacts with CBP/p300 and promotes hyperphosphorylation and degradation of the Notch ICD in vivo. Here we show that CycC:CDK8 and CycT1:CDK9/P-TEFb are recruited with Notch and associated coactivators (MAM, SKIP) to the HES1 promoter in signaling cells. MAM interacts directly with CDK8 and can cause it to localize to subnuclear foci. Purified recombinant CycC:CDK8 phosphorylates the Notch ICD within the TAD and PEST domains, and expression of CycC:CDK8 strongly enhances Notch ICD hyperphosphorylation and PEST-dependent degradation by the Fbw7/Sel10 ubiquitin ligase in vivo. Point mutations affecting conserved Ser residues within the ICD PEST motif prevent hyperphosphorylation by CycC:CDK8 and stabilize the ICD in vivo. These findings suggest a role for MAM and CycC:CDK8 in the turnover of the Notch enhancer complex at target genes. PMID- 15546613 TI - Identification of promoters bound by c-Jun/ATF2 during rapid large-scale gene activation following genotoxic stress. AB - The NH2-terminal Jun kinases (JNKs) function in diverse roles through phosphorylation and activation of AP-1 components including ATF2 and c-Jun. However, the genes that mediate these processes are poorly understood. A model phenotype characterized by rapid activation of Jun kinase and enhanced DNA repair following cisplatin treatment was examined using chromatin immunoprecipitation with antibodies against ATF2 and c-Jun or their phosphorylated forms and hybridization to promoter arrays. Following genotoxic stress, we identified 269 genes whose promoters are bound upon phosphorylation of ATF2 and c-Jun. Binding did not occur following treatment with transplatin or the JNK inhibitor SP600125 or JNK-specific siRNA. Of 89 known DNA repair genes represented on the array, 23 are specifically activated by cisplatin treatment within 3-6 hr. Thus, the genotoxic stress response occurs at least partly via activation of ATF2 and c Jun, leading to large-scale coordinate gene expression dominated by genes of DNA repair. PMID- 15546614 TI - Regulation of the pap epigenetic switch by CpxAR: phosphorylated CpxR inhibits transition to the phase ON state by competition with Lrp. AB - Pap pili gene expression is controlled by a reversible OFF/ON phase switch that is orchestrated by binding of Lrp to pap pilin promoter proximal sites 1, 2, and 3 (OFF) or pap promoter distal sites 4, 5, and 6 (ON). Movement of Lrp between proximal and distal sites controls pap pilin transcription and is modulated by PapI and DNA adenine methylase. Here we show that activation of the environmentally responsive CpxAR two-component regulatory system inhibits Pap phase variation by generation of phosphorylated CpxR (CpxR-P). CpxR-P competes with Lrp for binding to both promoter proximal and distal pap DNA binding sites, inhibiting pap transcription in vitro and pili expression in vivo. In contrast to Lrp, CpxR-P is methylation insensitive and does not form DNA methylation patterns in vivo. CpxAR-dependent repression of pap transcription is also observed in response to alkaline growth conditions. These results provide insight into a mechanism for environmental control of epigenetically regulated gene expression. PMID- 15546615 TI - A mechanism for inhibiting the SUMO pathway. AB - The SUMO pathway parallels the classical ubiquitinylation pathway with three discrete steps: activation involving the enzyme E1, conjugation involving the E2 enzyme UBC9, and substrate modification through the cooperative association of UBC9 and E3 ligases. We report here that the adenoviral protein Gam1 inhibits the SUMO pathway by interfering with the activity of E1 (SAE1/SAE2). In vivo, Gam1 expression leads to SAE1/SAE2 inactivation, both SAE1/SAE2 and UBC9 disappearance, and overall inhibition of protein sumoylation. This results in transcriptional activation of some promoters and is directly linked to inhibition of sumoylation of the transcriptional activators involved. Our results identify a mechanism for interfering with the SUMO pathway and with transcription that could have an impact in the design of novel pharmaceutical agents. They also point out once again to the extraordinary ability of eukaryotic viruses to interfere with the biology of host cells by targeting fundamental biochemical processes. PMID- 15546616 TI - Structure and function of an unusual family of protein phosphatases: the bacterial chemotaxis proteins CheC and CheX. AB - In bacterial chemotaxis, phosphorylated CheY levels control the sense of flagella rotation and thereby determine swimming behavior. In E. coli, CheY dephosphorylation by CheZ extinguishes the switching signal. But, instead of CheZ, many chemotactic bacteria contain CheC, CheD, and/or CheX. The crystal structures of T. maritima CheC and CheX reveal a common fold unlike that of any other known protein. Unlike CheC, CheX dimerizes via a continuous beta sheet between subunits. T. maritima CheC, as well as CheX, dephosphorylate CheY, although CheC requires binding of CheD to achieve the activity of CheX. Structural analyses identified one conserved active site in CheX and two in CheC; mutations therein reduce CheY-phosphatase activity, but only mutants of two invariant asparagine residues are completely inactive even in the presence of CheD. Our structures indicate that the flagellar switch components FliY and FliM resemble CheC more closely than CheX, but attribute phosphatase activity only to FliY. PMID- 15546617 TI - 20S proteasome differentially alters translation of different mRNAs via the cleavage of eIF4F and eIF3. AB - The molecular basis for coordinated regulation of protein synthesis and degradation is not understood. Here we report that the 20S proteasome endoproteolytically cleaves the translation initiation factors eIF4G, a subunit of eIF4F, and eIF3a, a subunit of eIF3. The cleavage of eIF4G or eIF3a differentially affects the assembly of ribosomal preinitiation complexes on different cellular and viral mRNAs in an in vitro system containing pure components. Inhibition of proteolytic activity of the 20S proteasome with specific inhibitors prevents cleavage of both factors in vitro and in vivo, restores assembly of ribosomal complexes in vitro, and differentially affects translation of different mRNAs in vivo. These studies demonstrate the importance of the endoproteolytic activity of proteasomes in regulation of cellular processes and suggest a link between protein synthesis and degradation. PMID- 15546618 TI - SMG7 acts as a molecular link between mRNA surveillance and mRNA decay. AB - Nonsense-mediated mRNA decay (NMD) is a surveillance mechanism that eliminates mRNAs containing premature termination codons (PTCs). The proteins UPF1, SMG5, SMG6, and SMG7 are essential NMD factors in metazoa. SMG5 and SMG7 form a complex with UPF1 and interact with each other via their N-terminal domains. Here we show that SMG5 and SMG7 colocalize in cytoplasmic mRNA decay bodies, while SMG6 forms separate cytoplasmic foci. When SMG7 is tethered to a reporter transcript, it elicits its degradation, bypassing the requirement for a PTC, UPF1, SMG5, or SMG6. This activity is mediated by the C-terminal domain of SMG7. In contrast, SMG5 requires SMG7 to trigger mRNA decay and to localize to decay bodies. Our findings indicate that SMG7 provides a link between the NMD and the mRNA degradation machinery by interacting with SMG5 and UPF1 via its N-terminal domain and targeting bound transcripts for decay via its C-terminal domain. PMID- 15546619 TI - Involvement of SR proteins in mRNA surveillance. AB - Nonsense mutations influence several aspects of gene expression, including mRNA stability and splicing fidelity, but the mechanism by which premature termination codons (PTCs) can apparently affect splice-site selection remains elusive. We used a model human beta-globin gene with duplicated 5' splice sites (5'ss) and found that PTCs inserted between the two 5'ss do not directly influence splicing in this system. Instead, their apparent effect on 5'ss selection in vivo is an indirect result of nonsense-mediated mRNA decay (NMD), as conditions that eliminated NMD also abrogated the effect on splicing. Remarkably, we found an unexpected function of SR proteins in targeting several mRNAs with PTCs to the NMD pathway. Overexpression of various SR proteins strongly enhanced NMD, and this effect required an RS domain. Our data argue against a universal role of PTCs in regulating pre-mRNA splicing and reveal an additional function of SR proteins in eukaryotic gene expression. PMID- 15546620 TI - Insights into strand displacement and processivity from the crystal structure of the protein-primed DNA polymerase of bacteriophage phi29. AB - The DNA polymerase from phage phi29 is a B family polymerase that initiates replication using a protein as a primer, attaching the first nucleotide of the phage genome to the hydroxyl of a specific serine of the priming protein. The crystal structure of phi29 DNA polymerase determined at 2.2 A resolution provides explanations for its extraordinary processivity and strand displacement activities. Homology modeling suggests that downstream template DNA passes through a tunnel prior to entering the polymerase active site. This tunnel is too small to accommodate double-stranded DNA and requires the separation of template and nontemplate strands. Members of the B family of DNA polymerases that use protein primers contain two sequence insertions: one forms a domain not previously observed in polymerases, while the second resembles the specificity loop of T7 RNA polymerase. The high processivity of phi29 DNA polymerase may be explained by its topological encirclement of both the downstream template and the upstream duplex DNA. PMID- 15546621 TI - An SMC-domain protein in fission yeast links telomeres to the meiotic centrosome. AB - Abnormal centrosomal structures similar to those occurring in human cancers are induced in fission yeast by overexpression of the pericentrin homolog Pcp1p. Analysis of abnormal Pcp1p-containing structures with quantitative mass spectrometry and isotope-coded affinity tags identified a coiled-coil, structural maintenance of chromosomes (SMC) domain protein. This protein, termed Ccq1p (coiled-coil protein quantitatively enriched), localizes with Taz1p to telomeres in normal vegetative cells. Fluorescence resonance energy transfer (FRET) measurements indicate that Ccq1p also interacts with centrosomal Pcp1p in mating pheromone-stimulated cells containing centrosomally clustered telomeres. We provide evidence that the Ccq1p-Pcp1p interaction, while essential for meiosis, is deleterious when forced to occur during vegetative growth. Cells lacking one ccq1 allele exhibit a loss-of-function phenotype including abnormally long cell length, chromosome segregation failure, telomeric shortening, and defective telomeric clustering during meiotic prophase. Our data indicate a mechanism underlying meiotic chromosomal bouquet formation and suggest a recruitment model for supernumerary centrosome toxicity. PMID- 15546622 TI - The RING domain of Mdm2 mediates histone ubiquitylation and transcriptional repression. AB - Histone modifications play a pivotal role in regulating transcription and other chromatin-associated processes. In yeast, histone H2B monoubiquitylation affects gene silencing. However, mammalian histone ubiquitylation remains poorly understood. We report that the Mdm2 oncoprotein, a RING domain E3 ubiquitin ligase known to ubiquitylate the p53 tumor suppressor protein, can interact directly with histones and promote in vitro monoubiquitylation of histones H2A and H2B. Moreover, Mdm2 induces H2B monoubiquitylation in vivo. Endogenous Mdm2 is tethered in vivo, presumably via p53, to chromatin comprising the p53 responsive p21(waf1) promoter, and Mdm2 overexpression enhances protein ubiquitylation in the vicinity of a p53 binding site within that promoter. Moreover, when recruited to a promoter in the absence of p53, Mdm2 can repress transcription dependently on its RING domain, suggesting that its E3 activity contributes to repression. Histone ubiquitylation may thus constitute a novel mechanism of transcriptional repression by Mdm2, possibly underlying some of its oncogenic activities. PMID- 15546623 TI - Distinct contributions of histone H3 lysine 9 and 27 methylation to locus specific stability of polycomb complexes. AB - The Polycomb group of proteins (PcG) maintains stable epigenetic silencing of over 100 genes via PcG response elements (PREs). Here we investigate the relationship between Polycomb binding, transcriptional status, and histone H3 methylation at lysine 9 (H3K9Me) and 27 (H3K27Me) for over 30 PcG targets in Drosophila. We show that H3K9Me and H3K27Me have distinct distributions at different loci. Our data show that Polycomb binding and histone methylation at the promoter do not prevent strong transcriptional activity, and indicate instead that silencing requires methylation of both PRE and promoter. In addition, we show that trimethylated H3K9 and H3K27 peptides can compete Polycomb from polytene chromosomes, with different effects at different loci, which correlate with differences in methylation status and transcriptional activity. We use mathematical modeling to examine these data, and propose that weak Polycomb histone tail interactions enable PcG complexes to bind dynamically to chromatin, offering opportunities for regulation. PMID- 15546624 TI - H2A.Z alters the nucleosome surface to promote HP1alpha-mediated chromatin fiber folding. AB - Controlling the degree of higher order chromatin folding is a key element in partitioning the metazoan genome into functionally distinct chromosomal domains. However, the mechanism of this fundamental process is poorly understood. Our recent studies suggested that the essential histone variant H2A.Z and the silencing protein HP1alpha may function together to establish a specialized conformation at constitutive heterochromatic domains. We demonstrate here that HP1alpha is a unique chromatin binding protein. It prefers to bind to condensed higher order chromatin structures and alters the chromatin-folding pathway in a novel way to locally compact individual chromatin fibers without crosslinking them. Strikingly, both of these features are enhanced by an altered nucleosomal surface created by H2A.Z (the acidic patch). This shows that the surface of the nucleosome can regulate the formation of distinct higher order chromatin structures mediated by an architectural chromatin binding protein. PMID- 15546625 TI - Spb1p-directed formation of Gm2922 in the ribosome catalytic center occurs at a late processing stage. AB - rRNA molecules undergo extensive posttranscriptional modification, predominantly 2'-O-ribose methylation and pseudouridine formation, both of which are guided by the numerous small nucleolar RNAs in eukaryotes. Here, we describe an exception to this rule. The essential yeast nucleolar protein Spb1p is a site-specific rRNA methyltransferase modifying the universally conserved G2922 that is located within the A loop of the catalytic center of the ribosome. The equivalent position in bacteria is the docking site for aminoacyl-tRNA, and it is critical for translation. In sharp contrast to other 2'-O-methylriboses that are formed on the primary transcript, Gm2922 appears at a late processing stage, during the maturation of the 27S pre-rRNA. Thus, eukaryotes have maintained a site-specific enzyme to catalyze the methylation of a nucleotide that plays a crucial role in ribosome biogenesis and translation. PMID- 15546626 TI - Toxicity and bioavailability of atrazine and molinate to the freshwater shrimp (Paratya australiensis) under laboratory and simulated field conditions. AB - Acute (96-h) semistatic toxicity tests were conducted by exposing the freshwater shrimp, Paratya australiensis, to atrazine and molinate in laboratory water and in river water both with and without sediment. The median lethal concentrations (LC50) and 95% fiducial limits of atrazine for P. australiensis in laboratory water in the absence and presence of sediment were 9.9 (8.6-11.5) and 6.8 (5.4 8.5)mg/L, respectively, while the corresponding values in river water were 9.8 (8.5-11.2) and 6.5 (5.4-7.8)mg/L, respectively. For molinate, the LC50 values in laboratory water in the absence and presence of sediment were 9.2 (7.0-12.1) and 9.0 (6.8-12.0)mg/L, respectively and the corresponding values in river water were 8.7 (6.4-11.8) and 8.2 (6.6-10.2)mg/L, respectively. Neither the river water nor the presence of sediment significantly (P<0.05) reduced the bioavailability of either chemical to P. australiensis. This was unexpected, as studies with other aquatic organisms have shown that sediment significantly reduced the bioavailability of these chemicals. PMID- 15546627 TI - The effect of Zn2+ on glucose 6-phosphate dehydrogenase activity from Bufo arenarum toad ovary and alfalfa plants. AB - The effect of Zn2+ on glucose 6-phosphate dehydrogenase (G6PD) activity was monitored in samples from Bufo arenarum toad ovary and alfalfa plants, in the search for a possible new bioindicator able to detect levels of exposure through contaminated soils, and also to elucidate possible similarities between the enzyme from animal and plant tissues. The in vivo effect was evaluated after exposure of the toads to the metal in Ringer solution during 30 days and after 10 days of treatment in 6 weeks old plants, cultured under laboratory conditions. In vitro effects were measured in different extracts from control samples and partially purified enzyme from ovarian tissue as well as in different extracts from control alfalfa plants, by addition of the metal to the reaction mixture containing the enzyme. G6PD from toad ovary was noncompetitively inhibited by zinc both in vivo and in vitro, under all the experimental conditions studied. A kinetic analysis of the enzyme activity showed that the Michaelis-Menten constant (Km) was not modified, while maximal velocity (Vmax) decreased as the consequence of treatment. It was not possible to obtain a dose-response curve for the effects of Zn2+ on G6PD from alfalfa whole plants, measured in vivo or in vitro. Only leaf extracts evidenced a possible relationship between treatment with the metal and G6PD activity alteration. The results agree with a possible role for G6PD as a biomarker of effect and exposure to Zn2+ in B. arenarum ovarian tissue but not in alfalfa plants. PMID- 15546628 TI - A comparative analysis of the toxicity of eight common soil contaminants and their effects on drought tolerance in the collembolan Folsomia candida. AB - Previous exposure to contaminants can increase the susceptibility to drought stress in soil invertebrates. In the present study, the effects of eight common environmental contaminants on drought tolerance and reproduction were investigated using the collembolan Folsomia candida as a model organism. Four classes of chemicals were considered: polycyclic aromatic hydrocarbons (pyrene and fluorene), detergents (linear alkylbenzene sulfonate (LAS) and nonylphenol), insecticides (dimethoate and cypermethrin), and heavy metals (copper and cadmium). Previous exposure to nonylphenol, pyrene, and fluorene had significant effects on drought tolerance. LAS had a weak effect on drought tolerance, whereas the remaining chemicals had no such effects even at concentrations at which reproduction was considerably reduced. Reproduction and adult survival in F. candida were equally sensitive to dimethoate. However, for all other chemicals, reproduction was a considerably more sensitive parameter than adult survival. The results suggest that toxicants whose effects are associated with cell membrane function are the most potent with respect to effect on drought tolerance. PMID- 15546629 TI - Linking land use variables and invertebrate taxon richness in small and medium sized agricultural streams on a landscape level. AB - In this study the average numbers of invertebrate species across an arable landscape in central Germany (surveys from 15 years in 90 streams at 202 sites) were assessed for their correlation with environmental factors such as stream width, land use (arable land, forest, pasture, settlement), soil type, and agriculture-derived stressors. The stress originating from arable land was estimated by the factor "risk of runoff," which was derived from a runoff model (rainfall-induced surface runoff). Multivariate analysis explained 39.9% of the variance in species number, revealing stream width as the most important factor (25.3%), followed by risk of runoff (9.7%). The results showed that wider streams -with or without agricultural stressors--contained significantly higher species numbers than narrower streams. This can be explained by potentially more diverse in-stream structures leading to more habitats and niches. However, negative effects on species number owing to runoff from arable land could be distinguished from the effect of stream width: the number of species within each stream width class significantly decreased with increasing risk of runoff. Therefore the factor "risk of runoff" is considered to express a significant proportion of the variability in macroinvertebrate communities caused by stressors of agricultural origin. PMID- 15546630 TI - Effects of dietary Pb(II) and tributyltin on neotropical fish, Hoplias malabaricus: histopathological and biochemical findings. AB - Trahira (Hoplias malabaricus) used to investigate the effects of successive Pb(II) or tributyltin (TBT) dietary doses. After 70 days of acclimation, individuals were exposed to 21 microg Pbg(-1) or 0.3 microg TBTg(-1) (5-day intervals, 14 doses). Two experiments were conducted to investigate the histopathological effects (liver and kidney) and measure the cholinesterase activity (muscle and brain) after Pb(II) or TBT dietary doses. A number of morphological effects were observed in liver, including cytoskeleton disturbance, microautophagy of mitochondria, nuclear damage, and cell death. In kidney, necrosis area, increasing of the neutrophils cell number, changes in melano macrophage centers, and free macrophages were frequently registered after both Pb(II) and TBT exposures. The cholinesterase activity was inhibited in muscle after 14 doses of Pb(II), but no effects were found in individuals exposed to TBT. In summary, this work is the first to report detailed in vivo toxic effects in tropical fish, H. malabaricus, after dietary sublethal exposure to Pb(II) and TBT. PMID- 15546631 TI - Effects of waterborne iron overload and simulated winter conditions on acute physiological stress response of whitefish, Coregonus lavaretus. AB - Two-year-old whitefish (Coregonus lavaretus) were exposed for 30 days to episodic iron overload in iron-rich humic water (5%) supplemented with inorganic iron (5 mg FeL(-1)). Two parallel laboratory exposures were performed, one under conditions simulating winter and the other under conditions simulating spring. After exposure, some of the fish were subjected to acute handling stress in the form of a short air challenge to reveal possible modification of the primary and secondary stress responses. In whitefish sampled without additional handling, iron accumulated in the liver (under spring conditions) and gills (under winter and spring conditions); plasma catecholamine and beta-estradiol (both winter and spring groups) as well as blood hematocrit (winter group only) levels were depressed; blood glucose (winter group only) and red blood cell (RBC) Na+ levels (spring group only) were increased. In handled whitefish, liver glycogen phosphorylase (GPase), RBC, and blood glucose stress responses were impaired by the applied exposure conditions, which reflected natural iron-rich humic water. Exposure also removed some physiological effects of the applied ambient conditions: plasma catecholamines and beta-estradiol, gill Na+/K+ -ATPase, and RBC K+ concentration were not different in two iron-exposed fish groups, whereas there was a difference in reference fish. Thus, the physiological effects of this type of subchronic exposure, together with alterations in the acute stress response, can lead to incorrect conclusions being drawn from the results, if the effects of time-dependent stress response are ignored. In conclusion, waterborne iron overload may impair the optimal capacity of whitefish to carry out their normal physiological functions such as responding to external threats. PMID- 15546632 TI - Single and joint toxicity of chlorimuron-ethyl, cadmium, and copper acting on wheat Triticum aestivum. AB - Investigation of the toxicological effects of some agricultural pollutants on germination rate and on shoot and root elongation of wheat (Triticum aestivum) was carried out. Seeds of wheat were exposed to various concentrations of chlorimuron-ethyl with or without cadmium and copper addition. The inhibitory rates of seed germination and shoot and root elongation of wheat were calculated. Significant linear relationships between the root and shoot elongation and the concentration of chlorimuron-ethyl with or without copper or cadmium addition were observed. The interactive effects of the three pollutants on seed germination were not significant (P > 0.05), but on shoot and root elongation they were markedly significant (P < 0.01). When concentrations of added Cd2+ or Cu2+ reached the concentration under which the inhibitory rate of wheat root elongation was about 20%, cadmium (or copper) and chlorimuron-ethyl had an antagonistic effect on the inhibition of shoot and root elongation, although the interactive effects of copper were not significant (P > 0.05). However, cadmium (or copper) and chlorimuron-ethyl had significantly synergic effects on the inhibition of shoot and root elongation (P < 0.05) when concentrations of added Cd2+ and Cu2+ reached the concentrations under which the inhibitory rate of root elongation was about 70%. At the high concentration of added cadmium or copper, joint toxicity of chlorimuron-ethyl and the heavy metals was more dependent on the concentrations of the latter. The sensitivity of wheat to the toxicity of the three pollutants was in the following sequence: root elongation>shoot elongation>germination rate. PMID- 15546633 TI - Hazard evaluation for northern bobwhite quail (Colinus virginianus) exposed to chlorpyrifos-treated turf and seed. AB - This study evaluated the toxicity effects of chlorpyrifos on bobwhite quail (Colinus virginianus) kept in 27 field-exposed large pens arranged over turf in a randomized block design with nine blocks of three pens (16 adult birds per pen). Nine pens were treated with one application of 3.4 kg active ingredient (ai) per hectare followed by a second 3.4 -kg ai/ha application 2 weeks later, nine pens with one 6.7 -kg ai/ha application, and nine pens with formulation blank. In addition, the seed fed to the birds in the two chemically treated pens was also treated with chlorpyrifos. Mean residue in the grass samples from the first 3.4 kg treatment pens ranged from 306+/-95 ppm on day 0 to 18+/-8 ppm on day 14 after treatment. The second 3.4 -kg ai/ha treatment grass residues ranged from 361+/ 167 ppm on day 0 to 38+/-24 ppm on day 14 after treatment. Grass residues from the 6.7-kg treatment pens ranged from 903+/-310 ppm on day 0 to 9+/-8 ppm on day 30 after treatment. Half-lives were approximately 2 days and 10 days for grass and seeds, respectively. Whereas the incidence of behavioral deficits was significantly (P = 0.0156) higher in the 6.7-kg pens (five females, one male), two of the females could have been the same bird because they were both seen in the same pen on days 23 and 24 after treatment. There was no significant difference in mortality, brain acetylcholinesterase activity, or any other measured parameter among any of the treatments. We conclude that application of chlorpyrifos to turf at 3.4 and 6.7 kg ai/ha is not expected to have chronic deleterious effects on populations of bobwhite quail grazing on treated grass or seeds, provided there is an abundant supply of seeds for the quail to eat. PMID- 15546634 TI - Toxicity of cypermethrin on growth, pigments, and superoxide dismutase of Scenedesmus obliquus. AB - Cypermethrin is a synthetic pyrethroid that is particularly toxic to crustaceans. It is therefore applied as a chemotherapeutant in farms for the treatment of pests. The effective concentrations of cypermethrin on the inhibition of Scenedesmus obliquus growth at 96 h (96 h EC50) were determined to be 50, 100, 150, 200, and 250 mg/L. Algal growth, pigment fractions, and the activity of superoxide dismutase (SOD) in the algal cells were measured in the exponential phase after exposure to cypermethrin. The results show that higher concentration of cypermethrin is inhibitory for growth and other metabolic activities and the 96 h EC50 of cypermethrin to S. obliquus is 112+/-9 mg/L; the potential application of SOD activity in S. obliquus as a sensitive biomarker for cypermethrin exposure is also discussed. PMID- 15546635 TI - Effects of the insecticide cypermethrin on plasma calcium and ultimobranchial gland of a teleost, Heteropneustes fossilis. AB - Heteropneustes fossilis were subjected to 5.76 and 1.44 microg/L of cypermethrin for short- and long-term experiments, respectively. Fish were sacrificed at 24, 48, 72, and 96 h in the short-term and 7, 14, 21, and 28 days in the long-term experiment. Plasma calcium levels were determined. Ultimobranchial glands were fixed for histological studies. Plasma calcium levels of fish exhibit a decrease after 48, 72, and 96 h. After 96 h a decrease in the staining response of the cytoplasm of ultimobranchial cells has been noticed. The nuclear volume of these cells undergoes a slight decrease. Chronically exposed fish exhibit a decrease in calcium level on day 7 which persists through 28 days. After 21 days, nuclear volume of the ultimobranchial cells undergoes a decrease and these cells exhibit a slight decrease in the staining response of the cytoplasm. Following 28 days exposure, the nuclear volume undergoes further decrease and degeneration and vacuolization sets in. PMID- 15546636 TI - Crude oil plus dispersant: always a boon or bane? AB - The toxicities of a Nigerian brand of crude oil (Forcados Light), a newly approved dispersant for use in Nigerian ecosystems (Biosolve), and their mixtures, based on ratios 9:1, 6:1 and 4:1 (v/v), were evaluated against the juvenile stage of prawn, Macrobrachium vollenhovenii, in laboratory bioassays. On the basis of the derived toxicity indices, crude oil with 96-h LC50 value of 0.28 ml/L was found to be about six times more toxic than the dispersant (96-h LC50 1.9 ml/L) when acting alone against M. vollenhovenii. Toxicity evaluations of the mixtures of crude oil and dispersant meant to simulate the environmental control settings of crude oil spillages in aquatic ecosystems revealed that effects of the crude oil/dispersant mixtures varied, depending largely upon the proportion of addition of the mixture components. The interactions between mixture of crude oil and dispersant at the test ratios of 9:1 and 4:1 were found to conform with the model of synergism (RTU=1.2 and 2.1, respectively), while the interactions between the mixture prepared based on ratio 6:1 conformed with the model of antagonism (RTU=0.16), based on the concentration-addition model. Furthermore, the mixtures prepared based on ratios 9:1 and 6:1 were found to be less toxic than crude oil when acting singly against M. vollenhovenii while the mixture prepared based on ratio 4:1 was found to have similar toxicity with crude oil when acting singly, based on the derived synergistic ratio values. The importance of results obtained from the joint-action tests in setting effective and environmentally safe dispersal ratios is discussed. PMID- 15546637 TI - Ecotoxicological evaluation of the biocidal agents sodium o-phenylphenol, sodium o-benzyl-p-chlorophenol, and sodium p-tertiary amylphenol. AB - The ecotoxicological effects of three biocidal agents frequently employed as active ingredients in phenolic-based disinfectants were evaluated using a test battery comprising of organisms representing three aquatic trophic levels. Phenolic-based disinfectants are commonly used by mushroom growers to disinfect spent mushroom compost. In general, the most sensitive assay used in this study was the Microtox test. In the case of the fish lethality assay, sodium o-benzyl-p chlorophenol was found to be slightly more sensitive than the bacterial test system. The freshwater alga and invertebrate tests were also among the most sensitive test species employed. The active ingredient, sodium o-benzyl-p chlorophenol (with the exception of the Microtox assay), was the most toxic chemical tested on each species. The majority of ecotoxicity data obtained in this research has not been previously reported and may therefore assist in the management and planning decisions regarding the application of pesticides and utilisation of SMC. PMID- 15546638 TI - The potential risk from 222radon posed to archaeologists and earth scientists: reconnaissance study of radon concentrations, excavations, and archaeological shelters in the Great Cave of Niah, Sarawak, Malaysia. AB - This reconnaissance study of radon concentrations in the Great Cave of Niah in Sarawak shows that in relatively deep pits and trenches in surficial deposits largely covered by protective shelters with poor ventilation, excavators are working in a micro-environment in which radon concentrations at the ground surface can exceed those of the surrounding area by a factor of > x 2. Although radon concentrations in this famous cave are low by world standards (alpha track etch results ranging from 100 to 3075 Bq m(-3)), they still may pose a health risk to both excavators (personal dosemeter readings varied from 0.368 to 0.857 mSv for 60 days of work) and cave occupants (1 yr exposure at 15 h per day with an average radon level of 608 Bq m(-3) giving a dose of 26.42 mSv). The data here presented also demonstrate that there is considerable local variation in radon levels in such environments as these. PMID- 15546639 TI - Dust accumulation and leaf pigment content in vegetation near the national highway at Sambalpur, Orissa, India. AB - An investigation on the seasonal variation in dust accumulation on leaves and leaf pigment content of six plant species of mixed habits was carried out at the side of the National Highway (NH 6) at Sambalpur, Orissa, India. The plants selected for study were Pongamia pinnata, Tabernaemontana divaricata, Ipomea carnea, Ficus relogiosa, Ficus benghalensis, and Quisqualis indica. The observed trend of dust accumulation was in the order T. divaricata>I. carnea>P. pinnata>F. religiosa>F. benghalensis>Q. indica. One-way analysis of variance showed significant difference in dust accumulation among plant species (F1 = 4.674, P < 0.01) and between seasons (F2 = 9.240, P < 0.01). It was seen that dust load increases with increasing number of vehicles using the highway (major emission source). The result shows significant correlation (negative) between dust load and pigment content in summer and rainy season. PMID- 15546640 TI - Sex-specific time patterns of suicidal acts on the German railway system. An analysis of 4003 cases. AB - OBJECTIVE: To examine sex-specific time patterns of suicidal behaviour on railway tracks as basis of suicide preventive strategies. METHOD: Cases were derived from the National Central Registry of all person accidents on the German railway net (STABAG) between 1997 and 2002 satisfying the operational definition of suicidal behaviour and included sex, age, date, clock time and outcome of the incidence. RESULTS: Over the 6-year observation period, 4003 fatal and non-fatal suicidal incidences were documented. Male to female ratio was 2.70:1 (p<0.0001). The female subgroup was significantly older than the male subgroup (p<0.0001). The monthly distribution revealed a bimodal pattern (p=0.01), particularly in men younger than 65 years, with an excess risk in April and September. This circannual pattern attenuated in the second half of the observation period as shown by adjusted Poisson regression. Monday and Tuesday proved to be high risk days for both sexes. For males and females, a bimodal diurnal distribution pattern with a morning and an evening peak was observed. While both sexes followed the same pattern in the winter half year, in summer females showed a pronounced excess risk in the morning hours while for men the evening peak was substantially amplified (p<0.0001). Risk assessment revealed a marked broadening by approximately 6 h of the time window in the summer half year compared to the winter half year. LIMITATIONS: About 30% of cases were excluded because of missing data on sex specification. However, cases with and without sex information did not differ significantly in the variables under investigation. CONCLUSION: The analysis revealed marked seasonal, weekly and diurnal peaks of railway suicide intensity. Differences between men and women indicate sex specific processes underlying their suicidal behaviour. The findings may increase alertness of railway and security personal for particular vulnerable time windows of excess risk for railway suicides. PMID- 15546641 TI - Cycloid psychoses are not part of a bipolar affective spectrum: results of a controlled family study. AB - BACKGROUND: Whereas a growing body of evidence suggests that cycloid psychoses have to be separated from schizophrenic psychoses, their relations to bipolar affective disorder are less clear. To further clarify this issue a controlled family study was undertaken. METHODS: All living and traceable adult first-degree relatives of 45 cycloid psychotic, 32 manic-depressive and 27 control probands were personally examined by an experienced psychiatrist blind to the diagnosis of the index proband. Data about not traceable relatives were collected by the "Family-History"-Method. A catamnestic diagnosis was established for each of the 431 relatives blind to family data. Age-corrected morbidity risks were calculated using the life-table method. RESULTS: Relatives of cycloid psychotic patients showed a significantly lower morbidity risk for endogenous psychoses in general and manic-depressive illness compared to relatives of patients with manic depressive illness. The familial morbidity risk for cycloid psychoses was low and did not differ significantly in both proband groups. Relatives of cycloid psychotic patients however did not differ significantly from relatives of controls regarding familial morbidity. LIMITATIONS: Our time-consuming methodical procedure implicated a relatively small number of participants due to restricted personnel resources. The restriction to hospitalised probands could possibly cause a limited representativity of the study sample. CONCLUSIONS: Our results suggest that cycloid psychoses are aetiologically different from manic-depressive illness and could not be integrated into a spectrum of bipolar affective disorders. The findings provide further evidence for a nosological independence of cycloid psychoses. PMID- 15546642 TI - Affective modulation of the startle response in depression: influence of the severity of depression, anhedonia, and anxiety. AB - BACKGROUND: The amplitude of the startle reflex response is known to be influenced by the concomitant presentation of affect-toned material--if it is positive affect-toned, the reflex is inhibited, and if it is negative affect toned, the reflex is augmented. Abundant evidence demonstrates the utility of the affect-startle paradigm as a significant tool for measuring both positive and negative emotions. We applied this paradigm to study emotional reactivity in depression, particularly in relation to symptoms of depression, anhedonia, and anxiety. METHODS: Depressed patients (22) and controls (22) were shown a series of film clips, consisting of two clips with positive valence, two with negative valence, and two with relatively neutral valence. The startle response was measured in reaction to the acoustic startle-eliciting stimuli presented three times binaurally during each clip. RESULTS: Highly depressed and anhedonic patients, relative to controls, showed a reduced mood (self-ratings) and a lack of startle modulation in response to affective film clips whereas patients relatively low on depression/anhedonia displayed a reduced mood only with pleasant clips and a normal pattern of affective startle modulation. Anhedonia and depression were highly positively correlated but neither correlated with anxiety. Anxious patients displayed larger reflexes across all clips and showed a reduced mood modulation with pleasant, but not unpleasant, clips. LIMITATIONS: The large majority of patients was medicated with antidepressants which may have influenced the results. CONCLUSIONS. Reactivity to pleasant stimuli is diminished in patients suffering from low levels of depression and/or anhedonia, but reactivity even to unpleasant stimuli seems compromised at high levels of depression and/or anhedonia. Anxiety is associated with hyperstartle responding. PMID- 15546643 TI - Prevalence and risk-factors for depression in elderly Turkish and Moroccan migrants in the Netherlands. AB - BACKGROUND: Western societies host increasing number of elderly labour migrants from Turkey and Morocco. The article studied the prevalence of clinically significant depressive symptoms among elderly Turkish and Moroccan migrants compared with native Dutch elderly and if differences in prevalence rates were explained by known risk factors for depression and/or ethnic, migration-related factors. METHODS: 330 Turkish, 299 Moroccan, and 304 Dutch elderly (55-74 years) were interviewed (cross-sectionally) using the Center for Epidemiologic Depression Scale (CES-D). Potential risk factors included sex, income level, marital status, ethnic origin, chronic physical illnesses, limitations in daily functioning, migration and acculturation questions. RESULTS: The prevalence of self-reported depressive symptoms (CES-D>or=16) was very high in elderly migrants, 33.6% for Moroccan and 61.5% for Turkish elderly. The prevalence of depressive symptoms in the native Dutch sample was similar to earlier studies in the Netherlands and abroad: 14.5%. Among migrants education and income level was very low and they had a high number of physical limitations and chronic medical illnesses. This only explained part of the ethnic differences found. In all three samples, depressive symptoms were associated with sex, chronic physical illness and physical limitations. In multivariate analysis, ethnic origin was uniquely associated with the presence of clinically significant depressive symptoms. Only a small number of remigration and acculturation items were associated with depressive symptoms in bivariate analysis. CONCLUSIONS: The prevalence of clinically significant depressive symptoms among elderly migrants from Turkey and Morocco in the Netherlands is very high. Ethnicity was a strong independent risk factor. PMID- 15546644 TI - Patient preference for counselling predicts postpartum depression: a prospective 1-year follow up study in high-risk women. AB - BACKGROUND: Patient preferences have been associated with a positive effect of depression treatment. Little is known about patient preferences in at-risk samples. The aim of this study was to examine the role of patient preference for counselling in the occurrence of postpartum depression in high-risk women. METHOD: We conducted a prospective 1-year follow up study in two hospitals and four midwifery practices in The Netherlands. Participants were 90 pregnant women at high risk for postpartum depression: 45 high-risk women who preferred no counselling, 45 high-risk women who preferred counselling. Both groups received care as usual. The main outcome measure was clinical depression (Research Diagnostic Criteria) at 3, 6, and 12 months postpartum. RESULTS: Point-prevalence rates of clinical depression were significantly higher in high-risk women who preferred counselling compared with high-risk women who did not prefer counselling (24% versus 9%, P=0.048; 19% versus 5%, P=0.048, at 3 and 6 months postpartum, respectively). No significant difference was found at 12 months postpartum. Across the first-year postpartum, high-risk women who preferred counselling were at seven-fold increased risk for clinical depression (OR=7.7, 95% CI 1.7-33.8, P=0.007). CONCLUSIONS: Patient preference for counselling is an important predictor of postpartum depression in pregnant women at high risk for postpartum depression. Patient preferences may reflect validly a perceived need for intervention in high-risk women. This finding emphasises the need to take patient preference for counselling into account as an important variable to identify a high-risk population. PMID- 15546645 TI - Predictors for 2-year outcome of major depressive episode. AB - In this 2-year prospective study, we searched for predictive factors influencing the 2-year outcome of major depressive episodes. Demographic characteristics (age, gender, education, employment), illness-related variables (severity, age at onset, number and duration of previous episodes), personality characteristics (DSM-IV personality disorders, trait anxiety, coping style), life context factors (life events before and during the depressive episode, social support, social adjustment), and biological markers (dexamethasone suppression test, thyroid stimulating hormone levels) of 117 inpatients with major depressive episode were assessed. A structural equation model was used to test the proposed correlational structure of the relevant variables. The non-remission of the depressive symptoms by the end of a 6-week acute treatment phase was found to be the most relevant factor predicting sustained non-remission at the end of a 2-year follow-up period. At the end of the sixth week, the severity of depression depended on the level of social support and on the severity of depression at baseline. Among the baseline variables, anxious personality traits and a lower level of education predicted a high level of depressive symptoms at the end of the 2-year follow-up. Life events before and during the depressive episode, and the biological markers at baseline had no direct effect on the outcome. The rapid remission of the depressive symptoms is the most important predictor for the favorable long-term outcome of a depressive episode. Personality characteristics, social support and level of education,--interacting with each other--also play a significant role. PMID- 15546646 TI - Spatial processing of facial emotion in patients with unipolar depression: a longitudinal study. AB - BACKGROUND: In this study, a face-in-the-crowd task was applied to examine the spatial detection of facial emotion as a function of depression and comorbid anxiety in the course of a psychotherapeutic inpatient treatment. METHODS: Patients with unipolar depression (n=22) and normal controls (n=22) were tested twice, about 7 weeks apart, on a face-in-the-crowd task using displays of schematic faces. Half the patients were suffering from a comorbid anxiety disorder. RESULTS: From test 1 to test 2, depressivity, frequency of negative thoughts, and worrying of patients improved significantly. At both sessions, depressed patients, irrespective of the presence of comorbid anxiety disorders, showed no performance differences in the detection of negative faces compared to controls. However, depressed patients with but not those without comorbid anxiety disorders were slower in responding to positive faces than controls. Both patient groups were slower in responding to the neutral faces' condition than controls. CONCLUSIONS: Our data indicate a spatial processing deficit for positive facial expression in depressives with a comorbid anxiety disorder. This impairment, which appears to persist during remission, might be due to deficits in effortful visual search processes. PMID- 15546647 TI - Does type of first contact in depressive and bipolar disorders predict subsequent hospitalisation and risk of suicide? AB - BACKGROUND: Only a few studies have investigated how the type of first contact is associated with the risk of subsequent hospitalisation and the risk of committing suicide for patients with depressive or bipolar disorders. METHOD: All outpatients (patients in psychiatric ambulatories and community psychiatry centres) and in-patients (patients admitted during daytime or overnight to a psychiatric hospital) with a diagnosis of depressive or bipolar disorder at first contact ever in a period from 1995 to 1999 in Denmark were identified from the Danish Psychiatric Central Research Register (DPCRR). The risk of subsequent hospitalisation and the risk of suicide were compared according to type of first contact. RESULTS: The risk of subsequent hospitalisation was significantly increased for patients who were admitted to inpatient facilities during first contact compared to patients with outpatient treatment as their first contact. Patients with depressive disorder who were admitted also had increased risk of committing suicide eventually. LIMITATIONS: The diagnoses are clinician based. CONCLUSIONS: Patients referred to inpatient treatment have a poorer long-term prognosis than patients treated as outpatients. PMID- 15546648 TI - Identifying suicidal ideation among older adults in a general practice setting. AB - BACKGROUND: Up to 70% of older people who commit suicide consult a general practitioner (GP) in the month prior to their death. The purpose of this study was to identify the clinical and demographic characteristics of older adults who are contemplating suicide and are in contact with a GP. METHODS: We utilised a cross-sectional study to investigate the association between suicidal ideation and clinical/demographic variables of 504 consecutive patients aged 60 years or over, attending 1 of 54 randomly selected Western Australian GPs. Prior to their medical consultation, patients completed a self-report questionnaire, which included questions about suicidal ideation (Depressive Symptom Inventory Suicidality Subscale, DSI-SS), demographic factors, lifestyle factors, physical health and mental health, including depression (Center for Epidemiologic Studies Depression Scale, CES-D). RESULTS: Within our sample of older patients, 6.3% acknowledged current suicidal ideation. Multivariate analyses indicated that current suicidal ideation was strongly associated with being depressed at least occasionally during the previous week (OR=7.3, 95% CI=2.3-23.0), CES-D scores of 16 points or greater (OR=3.6, 95% CI=1.0-12.1), and a prior history of attempted suicide (OR=15.5, 95% CI=4.0-60.6). LIMITATIONS: Our results and conclusions are limited to suicidal ideation, and may not apply to suicidal behaviour. CONCLUSIONS: Depressive symptomatology is strongly associated with suicidal ideation in later life. Strategies that enhance GPs' identification and treatment of affective illness in older patients should have the greatest impact on suicide rates within this age group. PMID- 15546649 TI - Aberration of cholesterol level in first-onset bipolar I patients. AB - Cholesterol interacts with serotonin and it has been found to be associated with some clinical symptoms of mood disorders. There is a paucity of data on first onset bipolar patients and from Asian population. The total cholesterol (TC) level was examined in 25 bipolar I patients with a single manic episode (BPSM) and 30 controls. The TC level was significantly lower in the BPSM than in the controls. There were negative correlations between the Young mania rating scale (YMRS) scores and the pretreatment TC level in BPSM. This study suggests that the TC level can be changed after treatment in bipolar manic patients, although more studies involving different ethnic groups will be needed. PMID- 15546650 TI - Prevalence of social phobia in a clinical sample of drug dependent patients. AB - INTRODUCTION: Social phobia is among the most frequent psychiatric disorders and can be classified into two subtypes, nongeneralized and generalized. Whereas it significantly worsens the morbidity of comorbid substance abuse disorders, and it often is associated with reduced treatment responses, there is still lacking data on its prevalence in clinical populations of drug abusing patients. METHODS: The study sample consisted of 75 inpatients and 75 outpatients meeting DSM-IV criteria for drug dependence. Symptoms of social phobia were assessed with the French-language version of the Liebowitz Social Anxiety Scale (LSAS). RESULTS: Prevalence rate were 20% for the generalized subtype and 42.6% for the nongeneralized subtype. Gender difference emerged in the severity of fear, women reporting significantly greater fear relating to performance situations than men. CONCLUSIONS: An important proportion of patients with substance dependence present a comorbid generalized or nongeneralized social phobia. Early recognition of social phobia and adequate interventions is warranted for these patients in order to improve their treatment response with regard to quality of life and relapse prevention. PMID- 15546651 TI - Acupuncture: a promising treatment for depression during pregnancy. AB - BACKGROUND: Few medically acceptable treatments for depression during pregnancy are available. The aim of this randomized controlled pilot study was to determine whether acupuncture holds promise as a treatment for depression during pregnancy. METHODS: Sixty-one pregnant women with major depressive disorder and a 17-item Hamilton Rating Scale for Depression (HRSD17) score >or=14 were randomly assigned to one of three treatments, delivered over 8 weeks: an active acupuncture (SPEC, N=20), an active control acupuncture (NSPEC, N=21), and massage (MSSG, N=20). Acupuncture treatments were standardized, but individually tailored, and were provided in a double-blind fashion. Responders to acute phase treatment (HRSD17 score<14 and >or=50% reduction from baseline) continued the treatment they were initially randomized to until 10 weeks postpartum. RESULTS: Response rates at the end of the acute phase were statistically significantly higher for SPEC (69%) than for MSSG (32%), with an intermediate NSPEC response rate (47%). The SPEC group also exhibited a significantly higher average rate of reduction in BDI scores from baseline to the end of the first month of treatment than the MSSG group. Responders to the acute phase of all treatments combined had significantly lower depression scores at 10 weeks postpartum than nonresponders. LIMITATIONS: Generalizability is limited by the small sample and its relative homogeneity. CONCLUSION: Acupuncture holds promise for the treatment of depression during pregnancy. PMID- 15546653 TI - A top runner of the flagellar world has unexpectedly gone. PMID- 15546654 TI - Translocation of bacterial proteins--an overview. AB - Recent progress in the understanding of the nature of the extraordinary variety of protein translocation systems, mainly in Gram negative bacteria, is reviewed. This takes us from the insertion of proteins into the inner membrane via the sophisticated Sec apparatus, the lethal injection of Type III proteins into host cells and on to the beautiful machine that assembles the flagellum. Attempts are made to establish some order, some common principles that might explain the variety and the complexity of some systems. The fundamentals considered are the nature of different transport signals, the nature of translocons (a wide variety of inner membrane types, outer membrane translocons are more conserved), the process of docking to translocons, the role of chaperones and the folding of transported proteins, the energetics of translocation, and prospects for future advances. PMID- 15546655 TI - SRP-mediated protein targeting: structure and function revisited. AB - The signal recognition particle (SRP) and its membrane-bound receptor (SR) deliver membrane proteins and secretory proteins to the translocation channel in the plasma membrane (or the endoplasmic reticulum). The general outline of the SRP pathway is conserved in all three kingdoms of life. During the past decade, structure determination together with functional studies has brought our understanding of the SRP-mediated protein transport to an almost molecular level. An impressive amount of new information especially on the prokaryotic SRP is integrated into the current picture of the SRP pathway. PMID- 15546656 TI - Sec-translocase mediated membrane protein biogenesis. AB - Alpha-helical transmembrane proteins in bacteria are localized within the plasma membrane. The membrane assembly of these proteins requires protein devices for insertion into the lipid bilayer. In E. coli, membrane proteins require the SRP pathway components Ffh, 4.5S RNA and FtsY for membrane targeting and the SecYEGDF translocase and, in some cases, SecA, for translocation of hydrophilic domains. In addition, YidC, a recently discovered membrane protein, mediates the membrane integration and folding of hydrophobic domains of membrane proteins. In this review, we will describe the current status of the protein targeting and membrane integration pathways. PMID- 15546657 TI - Membrane integration of E. coli model membrane proteins. AB - The molecular events of membrane translocation and insertion have been investigated using a number of different model proteins. Each of these proteins has specific features that allow interaction with the membrane components which ensure that the proteins reach their specific local destination and final conformation. This review will give an overview on the best-characterized proteins studied in the bacterial system and emphasize the distinct aspects of the pathways. PMID- 15546658 TI - Structure and function of SecA, the preprotein translocase nanomotor. AB - Most secretory proteins that are destined for the periplasm or the outer membrane are exported through the bacterial plasma membrane by the Sec translocase. Translocase is a complex nanomachine that moves processively along its aminoacyl polymeric substrates effectively pumping them to the periplasmic space. The salient features of this process are: (a) a membrane-embedded "clamp" formed by the trimeric SecYEG protein, (b) a "motor" provided by the dimeric SecA ATPase, (c) regulatory subunits that optimize catalysis and (d) both chemical and electrochemical metabolic energy. Significant recent strides have allowed structural, biochemical and biophysical dissection of the export reaction. A model incorporating stepwise strokes of the translocase nanomachine at work is discussed. PMID- 15546659 TI - The protein-conducting channel SecYEG. AB - In bacteria, the translocase mediates the translocation of proteins into or across the cytosolic membrane. It consists of a membrane embedded protein conducting channel and a peripherally associated motor domain, the ATPase SecA. The channel is formed by SecYEG, a multimeric protein complex that assembles into oligomeric forms. The structure and subunit composition of this protein conducting channel is evolutionary conserved and a similar system is found in the endoplasmic reticulum of eukaryotes and the cytoplasmic membrane of archaea. The ribosome and other membrane proteins can associate with the protein-conducting channel complex and affect its activity or functionality. PMID- 15546660 TI - The role of lipids in membrane insertion and translocation of bacterial proteins. AB - Phospholipids are essential building blocks of membranes and maintain the membrane permeability barrier of cells and organelles. They provide not only the bilayer matrix in which the functional membrane proteins reside, but they also can play direct roles in many essential cellular processes. In this review, we give an overview of the lipid involvement in protein translocation across and insertion into the Escherichia coli inner membrane. We describe the key and general roles that lipids play in these processes in conjunction with the protein components involved. We focus on the Sec-mediated insertion of leader peptidase. We describe as well the more direct roles that lipids play in insertion of the small coat proteins Pf3 and M13. Finally, we focus on the role of lipids in membrane assembly of oligomeric membrane proteins, using the potassium channel KcsA as model protein. In all cases, the anionic lipids and lipids with small headgroups play important roles in either determining the efficiency of the insertion and assembly process or contributing to the directionality of the insertion process. PMID- 15546661 TI - Catalysis of disulfide bond formation and isomerization in the Escherichia coli periplasm. AB - Disulfide bond formation is a catalyzed process in vivo. In prokaryotes, the oxidation of cysteine pairs is achieved by the transfer of disulfides from the highly oxidizing DsbA/DsbB catalytic machinery to substrate proteins. The oxidizing power utilized by this system comes from the membrane-embedded electron transport system, which utilizes molecular oxygen as a final oxidant. Proofreading of disulfide bond formation is performed by the DsbC/DsbD system, which has the ability to rearrange non-native disulfides to their native configuration. These disulfide isomerization reactions are sustained by a constant supply of reducing power provided by the cytoplasmic thioredoxin system, utilizing NADPH as the ultimate electron source. PMID- 15546662 TI - Quality control in the bacterial periplasm. AB - Studies of the mechanisms that Gram-negative bacteria use to sense and respond to stress have led to a greater understanding of protein folding in both cytoplasmic and extracytoplasmic locations. In response to stressful conditions, bacteria induce a variety of stress response systems, examples of which are the sigma(E) and Cpx systems in Escherichia coli. Induction of these stress response systems results in upregulation of several gene targets that have been shown to be important for protein folding under normal conditions. Here we review the identification of stress response systems and their corresponding gene targets in E. coli. In addition, we discuss the apparent redundancy of the folding factors in the periplasm, and we consider the potential importance of the functional overlap that exists. PMID- 15546663 TI - Tat-dependent protein targeting in prokaryotes and chloroplasts. AB - The twin-arginine translocation (Tat) system operates in the chloroplast thylakoid and the plasma membranes of a wide range of bacteria. It recognizes substrates bearing cleavable signal peptides in which a twin-arginine motif almost invariably plays a key role in recognition by the translocation machinery. These signal peptides are surprisingly similar to those used to specify transport by Sec-type systems, but the Tat pathway differs in fundamental respects from Sec type and other protein translocases. Its key attribute is its ability to translocate large, fully folded (even oligomeric) proteins across tightly sealed membranes. To date, three key tat genes have been characterised and the first details of the Tat system are beginning to emerge. In this article we review the salient features of Tat systems, with an emphasis on the targeting signals involved, the substrate specificities of Tat systems, our current knowledge of Tat complex structures and the known mechanistic features. Although the article is focused primarily on bacterial systems, we incorporate relevant aspects of plant thylakoid Tat work and we discuss how the plant and bacterial systems may differ in some respects. PMID- 15546664 TI - Type I secretion in gram-negative bacteria. AB - In gram-negative bacteria, type I secretion is carried out by a translocator made up of three proteins that span the cell envelope. One of these proteins is a specific outer membrane protein (OMP) and the other two are cytoplasmic membrane proteins: an ATP-binding cassette (ABC) and the so-called membrane fusion or adaptor protein (MFP). Type I secretion is sec-independent and bypasses the periplasm. This widespread pathway allows the secretion of proteins of diverse sizes and functions via a C-terminal uncleaved secretion signal. This C-terminal secretion signal specifically recognizes the ABC protein, triggering the assembly of the functional trans-envelope complex. This report will mainly deal will recent data concerning the structure and assembly of the secretion complex as well as the effects and role of substrate folding on secretion by this pathway. PMID- 15546665 TI - The underlying mechanisms of type II protein secretion. AB - The cell envelope of Gram-negative bacteria is composed of two membranes, which are separated by the peptidoglycan-containing periplasm. Whereas the envelope forms an essential barrier against harmful substances, it is nevertheless a compartment of intense traffic for large proteins such as enzymes and toxins. Numerous studies dealing with the molecular mechanism of protein secretion have revealed that Gram-negative bacteria evolved different strategies to achieve this process. Among them, the type II secretion mechanism is part of a two-step process. Exoproteins following this pathway are synthesized as signal peptide containing precursors. After cleavage of the signal peptide, the mature exoproteins are released into the periplasm, where they fold. The type II machinery, also known as the secreton, is responsible for the translocation of the periplasmic intermediates across the OM. The type II system is broadly conserved in Gram-negative bacteria and involves a set of 12-16 different proteins named GspC-M, GspAB, GspN, GspO, and GspS. The type II secretion system is highly reminiscent of the type IV piliation assembly system. Based on findings about the subcellular localisation of the Gsp components, protein-protein interactions between Gsps and their multimerisation status, structural data and electron microscopy observation, it could be proposed a working model that strikingly runs both systems in parallel. PMID- 15546666 TI - Type III protein secretion mechanism in mammalian and plant pathogens. AB - The type III protein secretion system (TTSS) is a complex organelle in the envelope of many Gram-negative bacteria; it delivers potentially hundreds of structurally diverse bacterial virulence proteins into plant and animal cells to modulate host cellular functions. Recent studies have revealed several basic features of this secretion system, including assembly of needle/pilus-like secretion structures, formation of putative translocation pores in the host membrane, recognition of N-terminal/5' mRNA-based secretion signals, and requirement of small chaperone proteins for optimal delivery and/or expression of effector proteins. Although most of our knowledge about the TTSS is derived from studies of mammalian pathogenic bacteria, similar and unique features are learned from studies of plant pathogenic bacteria. Here, we summarize the most salient aspects of the TTSS, with special emphasis on recent findings. PMID- 15546667 TI - Type III flagellar protein export and flagellar assembly. AB - Bacterial flagella, unlike eukaryotic flagella, are largely external to the cell and therefore many of their subunits have to be exported. Export is ATP-driven. In Salmonella, the bacterium on which this chapter largely focuses, the apparatus responsible for flagellar protein export consists of six membrane components, three soluble components and several substrate-specific chaperones. Other flagellated eubacteria have similar systems. The membrane components of the export apparatus are housed within the flagellar basal body and deliver their substrates into a channel or lumen in the nascent structure from which point they diffuse to the far end and assemble. Both on the basis of sequence similarities of several components and structural similarities, the flagellar protein export systems clearly belong to the type III superfamily, whose other members are responsible for secretion of virulence factors by many species of pathogenic bacteria. PMID- 15546669 TI - Protein secretion through autotransporter and two-partner pathways. AB - Two distinct protein secretion pathways, the autotransporter (AT) and the two partner secretion (TPS) pathways are characterized by their apparent simplicity. Both are devoted to the translocation across the outer membrane of mostly large proteins or protein domains. As implied by their name, AT proteins contain their own transporter domain, covalently attached to the C-terminal extremity of the secreted passenger domain, while TPS systems are composed of two separate proteins, with TpsA being the secreted protein and TpsB its specific transporter. In both pathways, the secreted proteins are exported in a Sec-dependent manner across the inner membrane, after which they cross the outer membrane with the help of their cognate transporters. The AT translocator domains and the TpsB proteins constitute distinct families of protein-translocating, outer membrane porins of Gram-negative bacteria. Both types of transporters insert into the outer membrane as beta-barrel proteins possibly forming oligomeric pores in the case of AT and serve as conduits for their cognate secreted proteins or domains across the outer membrane. Translocation appears to be folding-sensitive in both pathways, indicating that AT passenger domains and TpsA proteins cross the periplasm and the outer membrane in non-native conformations and fold progressively at the cell surface. A major difference between AT and TPS pathways arises from the manner by which specificity is established between the secreted protein and its transporter. In AT, the covalent link between the passenger and the translocator domains ensures the translocation of the former without the need for a specific molecular recognition between the two modules. In contrast, the TPS pathway has solved the question of specific recognition between the TpsA proteins and their transporters by the addition to the TpsA proteins of an N proximal module, the conserved TPS domain, which represents a hallmark of the TPS pathway. PMID- 15546670 TI - Fiber assembly by the chaperone-usher pathway. AB - Bacterial pathogens utilize the chaperone-usher pathway to assemble extracellular multi-subunit fibers essential for virulence. The periplasmic chaperone facilitates the initial folding of fiber subunits but then traps them in activated folding transition states. Chaperone dissociation releases the folding energy that drives subunit incorporation into the fiber, which grows through a pore formed by the outer-membrane usher. PMID- 15546668 TI - Type IV secretion: the Agrobacterium VirB/D4 and related conjugation systems. AB - The translocation of DNA across biological membranes is an essential process for many living organisms. In bacteria, type IV secretion systems (T4SS) are used to deliver DNA as well as protein substrates from donor to target cells. The T4SS are structurally complex machines assembled from a dozen or more membrane proteins in response to environmental signals. In Gram-negative bacteria, the conjugation machines are composed of a cell envelope-spanning secretion channel and an extracellular pilus. These dynamic structures (i) direct formation of stable contacts-the mating junction-between donor and recipient cell membranes, (ii) transmit single-stranded DNA as a nucleoprotein particle, as well as protein substrates, across donor and recipient cell membranes, and (iii) mediate disassembly of the mating junction following substrate transfer. This review summarizes recent progress in our understanding of the mechanistic details of DNA trafficking with a focus on the paradigmatic Agrobacterium tumefaciens VirB/D4 T4SS and related conjugation systems. PMID- 15546671 TI - Protein sorting to the cell wall envelope of Gram-positive bacteria. AB - The covalent anchoring of surface proteins to the cell wall envelope of Gram positive bacteria occurs by a universal mechanism requiring sortases, extracellular transpeptidases that are positioned in the plasma membrane. Surface protein precursors are first initiated into the secretory pathway of Gram positive bacteria via N-terminal signal peptides. C-terminal sorting signals of surface proteins, bearing an LPXTG motif or other recognition sequences, provide for sortase-mediated cleavage and acyl enzyme formation, a thioester linkage between the active site cysteine residue of sortase and the C-terminal carboxyl group of cleaved surface proteins. During cell wall anchoring, sortase acyl enzymes are resolved by the nucleophilic attack of peptidoglycan substrates, resulting in amide bond formation between the C-terminal end of surface proteins and peptidoglycan cross-bridges within the bacterial cell wall envelope. The genomes of Gram-positive bacteria encode multiple sortase genes. Recent evidence suggests that sortase enzymes catalyze protein anchoring reactions of multiple different substrate classes with different sorting signal motif sequences, protein linkage to unique cell wall anchor structures as well as protein polymerization leading to the formation of pili on the surface of Gram-positive bacteria. PMID- 15546672 TI - Type I signal peptidases of Gram-positive bacteria. AB - Proteins that are exported from the cytoplasm to the periplasm and outer membrane of Gram-negative bacteria, or the cell wall and growth medium of Gram-positive bacteria, are generally synthesized as precursors with a cleavable signal peptide. During or shortly after pre-protein translocation across the cytoplasmic membrane, the signal peptide is removed by signal peptidases. Importantly, pre protein processing by signal peptidases is essential for bacterial growth and viability. This review is focused on the signal peptidases of Gram-positive bacteria, Bacillus and Streptomyces species in particular. Evolutionary concepts, current knowledge of the catalytic mechanism, substrate specificity requirements and structural aspects are addressed. As major insights in signal peptidase function and structure have been obtained from studies on the signal peptidase LepB of Escherichia coli, similarities and differences between this enzyme and known Gram-positive signal peptidases are highlighted. Notably, while the incentive for previous research on Gram-positive signal peptidases was largely based on their role in the biotechnologically important process of protein secretion, present-day interest in these essential enzymes is primarily derived from the idea that they may serve as targets for novel anti-microbials. PMID- 15546673 TI - Bacillus subtilis as cell factory for pharmaceutical proteins: a biotechnological approach to optimize the host organism. AB - Bacillus subtilis is a rod-shaped, Gram-positive soil bacterium that secretes numerous enzymes to degrade a variety of substrates, enabling the bacterium to survive in a continuously changing environment. These enzymes are produced commercially and this production represents about 60% of the industrial-enzyme market. Unfortunately, the secretion of heterologous proteins, originating from Gram-negative bacteria or from eukaryotes, is often severely hampered. Several bottlenecks in the B. subtilis secretion pathway, such as poor targeting to the translocase, degradation of the secretory protein, and incorrect folding, have been revealed. Nevertheless, research into the mechanisms and control of the secretion pathways will lead to improved Bacillus protein secretion systems and broaden the applications as industrial production host. This review focuses on studies that aimed at optimizing B. subtilis as cell factory for commercially interesting heterologous proteins. PMID- 15546674 TI - Post-translocational folding of secretory proteins in Gram-positive bacteria. AB - The transport of proteins from their site of synthesis in the cytoplasm to their functional location is an essential characteristic of all living cells. In Gram positive bacteria the majority of proteins that are translocated across the cytoplasmic membrane are delivered to the membrane-cell wall interface in an essentially unfolded form. They must then be folded into their native configuration in an environment that is dominated by a high density of immobilised negative charge-in essence an ion exchange resin. It is essential to the viability of the cell that these proteins do not block the translocation machinery in the membrane, form illegitimate interactions with the cell wall or, through intermolecular interactions, form insoluble aggregates. Native Gram positive proteins therefore have intrinsic folding characteristics that facilitate their rapid folding, and this is assisted by a variety of folding factors, including enzymes, peptides and metal ions. Despite these intrinsic and extrinsic factors, secretory proteins do misfold, particularly if the cell is subjected to certain types of stress. Consequently, Gram-positive bacteria such as Bacillus subtilis encode membrane- and cell wall-associated proteases that act as a quality control machine, clearing misfolded or otherwise aberrant proteins from the translocase and the cell wall. PMID- 15546675 TI - Estimating the safe starting dose in phase I clinical trials and no observed effect level based on QSAR modeling of the human maximum recommended daily dose. AB - Estimating the maximum recommended starting dose (MRSD) of a pharmaceutical for phase I human clinical trials and the no observed effect level (NOEL) for non pharmaceuticals is currently based exclusively on an extrapolation of the results of animal toxicity studies. This process is inexact and requires the results of toxicity studies in multiple species (rat, dog, and monkey) to identify the no observed adverse effect level (NOAEL) and most sensitive test species. Multiple uncertainty (safety) factors are also necessary to compensate for incompatibility and uncertainty underlying the extrapolation of animal toxicity to humans. The maximum recommended daily dose for pharmaceuticals (MRDD) is empirically derived from human clinical trials. The MRDD is an estimated upper dose limit beyond which a drug's efficacy is not increased and/or undesirable adverse effects begin to outweigh beneficial effects. The MRDD is essentially equivalent to the NOAEL in humans, a dose beyond which adverse (toxicological) or undesirable pharmacological effects are observed. The NOAEL in test animals is currently used to estimate the safe starting dose in human clinical trials. MDL QSAR predictive modeling of the human MRDD may provide a better, simpler and more relevant estimation of the MRSD for pharmaceuticals and the toxic dose threshold of chemicals in humans than current animal extrapolation based risk assessment models and may be a useful addition to current methods. A database of the MRDD for over 1300 pharmaceuticals was compiled and modeled using MDL QSAR software and E-state and connectivity topological descriptors. MDL QSAR MRDD models were found to have good predictive performance with 74-78% of predicted MRDD values for 120 internal and 160 external validation compounds falling within a range of +/-10-fold the actual MRDD value. The predicted MRDD can be used to estimate the MRSD for pharmaceuticals in phase I clinical trials with the addition of a 10 fold safety factor. For non-pharmaceutical chemicals any compound-related effect can be considered an undesirable and adverse toxicological effect and the predicted MRDD can be used to estimate the NOEL with the addition of an appropriate safety factor. PMID- 15546676 TI - Synchronous evaluation of toxico- and pharmaco-dynamics of yttrium-90 by a novel autoradiographic procedure. AB - A synchronous evaluation was performed, using a quick in vivo [2-(14)C]thymidine labeling method, of the toxico- and pharmaco-dynamics of a given dose of yttrium 90 (90Y) at a given time after injection to nude BALB/c mice loaded with 10(7) HuO9 cells. Quantitative data were 14C-microautoradiographs of the liver lobule, intestinal crypts, epiphysial growth plate, secondary ossification center containing pluripotent stem cells, perifollicular zone containing unipotent stem cells in the spleen, and plasmacytoma cells in the osteogenic sarcoma in each mouse following a 10-min labeling with 14C at 0.5, 6, and 24 h after i.v. injection of 90Y. Results show that the cell proliferation rate of the stem cells in respective tissues was markedly suppressed, dependent on time after dosing and the dose of 90Y; 3.7, 37, 370, 3700, and 37,000 kBq per mouse (25 g). In addition to the above, the sensitivity of the proliferation rate was dependent on amitosis or mitosis and the AUC value of 90Y-concentration at specific locations of the cells in the mouse body. The most sensitive cells were the plasmacytoma cells, followed by the pluripotent and unipotent stem cells, the intestinal crypts, epiphysial growth plate, and liver cells. PMID- 15546677 TI - Evaluation of a surrogate antibody for preclinical safety testing of an anti CD11a monoclonal antibody. AB - Surrogate antibodies are a potential solution to the limited safety testing possible with humanized monoclonal antibodies with restricted species cross reactivity. However, there are currently no defined criteria by which a potential surrogate antibody should be judged prior to its use in determining safety issues for the clinical agent. We propose that, potential surrogates should undergo rigorous evaluation to assess pharmacological and toxicological activities in comparison to the clinical agent. The current studies evaluated a chimeric mouse/rat anti-mouse CD11a monoclonal antibody (muM17) as a potential surrogate for efalizumab, a humanized anti-CD11a antibody in development for psoriasis. CD11a is a subunit of lymphocyte function associated antigen-1, an integrin involved in cell-cell interactions important to immune responses and inflammation. In vitro pharmacology studies included binding affinity to whole mouse blood and inhibitory activity of muM17 in a mixed lymphocyte response assay. In vivo pharmacology was examined using a delayed type hypersensitivity assay in female CD-1 mice. The toxicology evaluation included a murine tissue cross-reactivity study and in vivo multiple dose studies in female CD-1 mice which were administered muM17 (0.1-30 mg/kg) via subcutaneous injections once a week for 4 weeks. Clinical observations, body weight, clinical pathology, T cell CD11a expression, immunogenicity, toxicokinetics, and lymphoid organ histopathology were evaluated. Finally, since reproductive safety testing would be an important application of the proposed surrogate antibody, a pilot study in pregnant mice was conducted that demonstrated proportional transfer of muM17 into the fetus. These studies demonstrated that muM17 has pharmacological and toxicological activities similar to efalizumab. The selection of dose and regimen for GLP (Good Laboratory Practice) toxicology studies and extrapolation to clinical dose levels was based on pharmacodynamic activity (CD11a downmodulation on T cells). PMID- 15546678 TI - A risk-based methodology for deriving quality standards for organic contaminants in sewage sludge for use in agriculture--Conceptual Framework. AB - This paper describes a systematic methodology (Conceptual Framework) to derive quality standards for organic (anthropogenic) contaminants in sewage sludge added to agricultural land, in the context of revision of EU Sludge Directive 86/278/EEC and the broader Soil Thematic Strategy. The overall objective is to ensure, based on a risk assessment approach, a sustainable use of sludge over a long time horizon. ILSI-Europe's Conceptual Framework is in essence consistent with the EU Technical Guidance Document (TGD) for Environmental Risk Assessment of Chemicals in the soil compartment, or US-EPA's Sewage Sludge Use and Disposal Regulations, Part 503 Standards. A 'checklist' of different exposure pathways and transfer processes for organic contaminants needs to be considered, and the most sensitive relevant toxicological endpoint and its PNEC need to be identified. The additional complexity specific to deriving Sludge Quality Standards (SQS) is that the toxicity results may need-e.g., for (indirect) human toxicity-to be related back to maximum acceptable soil exposure levels (PEC(soil)). In turn, the latter need to be back-calculated to the maximum acceptable levels in sewage sludge (PEC(sludge)) at the time of application. Finally, for a sustainable sludge use, the exposure from repeated addition and potential chemical build-up over time (e.g., 100 years) needs to be assessed. The SQS may therefore vary with the (local) sludge application regime, and/or sludge pretreatment processes. PMID- 15546679 TI - Statistical methodology to evaluate food exposure to a contaminant and influence of sanitary limits: application to Ochratoxin A. AB - This paper presents some statistical methodologies to evaluate the food exposure to a contaminant and quantify the outcome of a new maximum limit on a food item. Our application deals with Ochratoxin A (OTA). We focus on the quantitative evaluation of the distribution of exposure based on both consumption data and contamination data. One specific aspect of contamination data is left censorship due to the limits of detection. Three calculation procedures are proposed: [P1] a deterministic method using means of contamination; [P2] a probabilistic method using a parametric adjustment of the distributions of contamination taking into account the left censorship; and [P3] a non-parametric method which consists in randomly selecting the consumption data and the contamination values. Our main result shows that a non-parametric probabilistic approach is well adapted for the purpose of exposure assessment, when large samples are available. In the application to OTA, the probability to exceed a safe level is high, particularly for children. Simulations show that the impact of the existing standards on cereals and the currently proposed standards on wine generally do not significantly reduce the risk to be overexposed to OTA. PMID- 15546680 TI - Magnitude and mechanistic determinants of the interspecies toxicokinetic uncertainty factor for organic chemicals. AB - The interspecies uncertainty factor, UF(AH), of 10 was recently subdivided into two components to account separately for interspecies differences in toxicokinetics and toxicodynamics (UF(AH-TK)=3.16, UF(AH-TD)=3.16). Even though the UF(AH) in its composite or dissociated form is used during the health risk assessment of systemic toxicants, there is no convincing scientific basis to justify the use of the same UF for all chemicals. In this study, we use equations that describe the toxicokinetics of chemicals at steady-state to characterize the magnitude and mechanistic determinants of UF(AH-TK) for several volatile organic chemicals (VOCs). Further, algorithms have been developed to determine the magnitude of the components of UF(AH-TK), namely the UF(AH-TK-ABS) (accounting for interspecies differences in dose absorbed during identical inhalation exposure conditions), UF(AH-TK-MET) (referring to the factor by which the blood concentration of unchanged parent chemical differs from one species to another, due to metabolic clearance, when both species receive identical doses) and UF(AH TK-DIS) (reflecting the magnitude of difference in chemical concentrations distributed in target tissues of two species when the arterial blood concentration in both species is identical). The results show that the body weight, the rate of alveolar ventilation, the fraction of cardiac output flowing to the liver, partition coefficients (blood:air and tissue:blood), and the hepatic extraction ratio are the only parameters that play a critical role in the extrapolation of tissue and blood concentrations across species. Further, the magnitude of the UF(AH-TK-ABS) (means+/-SD, 0.19+/-0.04), UF(AH-TK-MET) (means+/ SD, 0.24+/-0.05) and UF(AH-TK-DIS) (mean range: 1.76-0.93) obtained in this study for several VOCs compares well with that obtained previously using physiologically based toxicokinetic models. PMID- 15546681 TI - An intervention analysis for the reduction of exposure to methylmercury from the consumption of seafood by women of child-bearing age. AB - A previously developed exposure model was used [Risk Anal. 22 (2002) 689] to assess the effectiveness of various advisory scenarios on minimizing mercury (Hg) blood levels via the consumption of commercial seafood, both finfish and shellfish. This exposure model was developed to predict levels of Hg in blood in women of child-bearing age in the US based on the frequency of seafood consumption, the amount of seafood consumed per serving, and the types of seafood consumed. Steady-state relationships that employed descriptive statistics to account for toxicokinetic variation were used to predict levels of Hg in blood. The model incorporates an uncertainty dimension that is intended to represent the range of plausible interpretations of the data. The predictability of the model was confirmed via the use of National Health and Nutrition Examination Survey (NHANES) blood Hg data. In the present analysis, the model was used to predict the impact of limitations in the amount or types of seafood consumed on blood Hg levels. Specifically, simulations for various advisory scenarios were developed on the basis of limitations on total consumption of seafood, elimination of the consumption of certain species altogether, and/or a combination of both. In the baseline model, the median (uncertainty) estimates for the 50th, 95th, and 99th per capita population percentiles were 1.25, 8.2, and 16.1 ppb blood Hg, respectively. After restriction of seafood consumption to no more than 12 oz/week, the median (uncertainty) estimates for the 50th, 95th, and 99th per capita population percentiles were 1.22, 6.8, and 10.6 ppb blood Hg, respectively. Elimination of MeHg species, with average concentrations above 0.6 ppm, resulted in very modest decrements in Hg blood levels, in comparison to either the baseline or the reduced consumption scenarios. These results suggest that strategies to reduce MeHg exposure by reducing the amount of fish consumed (e.g., 12 oz/week) are more effective at eliminating the high end of the exposure distribution than are strategies intended to change the types of fish consumed. PMID- 15546682 TI - Toxicity evaluation of a beta-galactosidase preparation produced by Penicillium multicolor. AB - Tilactase is a beta-galactosidase enzyme preparation having lactase activity produced from the fungus Penicillium multicolor. The safety of tilactase was investigated in rats, dogs, and rabbits. Adult and juvenile rats administered 0, 500, 1000, or 4000 mg/kg bw/day of the enzyme preparation by gavage for 35 days, and dogs administered 0, 200, 500, or 1000 mg/kg bw/day in capsules for 30 days, exhibited no significant dose-related changes in body weights, feed consumption, organ weights, urinalysis, hematological profiles, clinical chemistry, or histopathological profiles. Rats receiving the same doses for 6 months also exhibited no dose-related effects, except for a small increase in the weight of the large intestine, an effect considered to be a physiological reaction to passage of a large amount of a non-absorbable substance. The no-observable adverse-effect level (NOAEL) was 4000 mg/kg bw/day for rats and 1000 mg/kg bw/day for dogs. In three separate studies to examine reproductive and developmental toxicity, rats received 0, 250, 1000 or 4000 mg/kg bw/day by gavage up to the 7th day of pregnancy, during days 7-17 of pregnancy, and from day 17 of pregnancy to 21 days after delivery. There were no treatment-related effects on the dams, gestation period, numbers of implantations, parturition rates, sex ratios, or survival of offspring in any of the studies. No treatment-related external, internal, or skeletal abnormalities were observed in fetuses from any of the three studies. The NOAEL was 4000 mg/kg bw/day. In addition to the three rat studies, rabbits received 0, 250, 500, or 1000 mg/kg bw/day by gavage from the 6th to 18th day of pregnancy. No treatment-related changes were observed in the dams, or fertility indices; nor were there any treatment-related fetal abnormalities. The NOAEL was 1000 mg/kg bw/day. When viable P. multicolor spores were injected into the tail veins of mice, no deaths occured, no fungal cells were observed in various organs, and histopathology showed only focal necrosis in the liver of some of the animals, including the controls. Similar effects were observed when spores were administered to mice in a single dose by gavage. The particular strain of P. multicolor used to prepare tilactase is not pathogenic. PMID- 15546683 TI - The stability of historical control data for common neoplasms in laboratory rats and the implications for carcinogenic risk assessment. AB - Time-related changes in the incidences of spontaneous neoplasms in skin (fibroma and keratoacanthoma), thyroid (C-cell and follicular cell adenomas/carcinomas), uterus (stromal polyp), testes (Leydig cell tumor) and hemolymphoreticular system (mesenteric lymph node hemangioma and malignant granular lymphocytic leukemia) were assessed statistically in Wistar, Sprague-Dawley and F344 rats employed by the BASF, Germany and major European contract research organizations over the last 20 years. Negative trends (5 out of 80 cases) were observed for skin fibromas in F344 males, for follicular cell adenomas in Han Wistar females and in Sprague-Dawley males and females, and for follicular cell carcinomas in Sprague Dawley males. Positive trends (8 out of 80 cases) were observed for skin keratoacanthomas in Han Wistar males, for C-cell adenomas in BASF Wistar males and females, for stromal polyps in Han Wistar and Sprague-Dawley females, and for mesenteric lymph node hemangiomas in Han Wistar and Sprague-Dawley males and in BASF Wistar females. In 67 out of 80 cases there were no statistically significant trends. Tumor drift was not common but occurred far more often in outbred rat strains (Wistar and Sprague-Dawley) than in the inbred rat strain (F344). This observation suggests that tumor predisposition is genetically determined, that tumor drift is primarily caused by genetic drift and that non genotoxic carcinogens operate by facilitating the expression of tumor predisposition in target cells. PMID- 15546684 TI - Toxicological resources for cumulative risk: an example with hazardous air pollutants. AB - Cumulative risk assessment, concerned with the multiple health effects of chemical mixtures, challenges the utility of existing single-chemical regulatory references. We compare example cumulative risk assessments for 40 HAPs; one based on single-effect toxicological data from EPA, and another based on a multiple effect toxicological database we developed. For the 40-chemical HAP subset, the multiple effect database contains information on approximately seven effects per chemical and contains a total of 290 toxicological values. Seven health effects are represented in the IRIS data. Seventeen health effects are represented in the multiple-effect data. Respiratory and neurological effects ranked first and second in both cumulative analyses, regardless of the source data. In addition to respiratory and neurological effects, gastro-intestinal/hepatic, renal/kidney, and immunologic effects were identified as effects of concern on the basis of the multiple effect data. Immunologic effects are not found in the 40-chemical IRIS dataset. Cumulative risk assessment has the potential to expand our understanding of the public health impacts of environmental exposures. Advancements in toxicological resources will improve cumulative risk assessment. Cumulative risk assessment will reduce risks to the extent that it can be integrated into prevention strategies to track and protect the public's health. PMID- 15546685 TI - Improving protection for research subjects in France: analysis of regional ethics committees. AB - OBJECTIVE: The aim of this study was to analyze the functioning of the French committees for the protection of people in biomedical research (CCPPRB). DESIGN: An independent evaluator visited all of the committees and analyzed their functioning by assessing the annual activity reports, filling data grids and observing the committees in session. RESULTS: We observed that the representation of a range of professions, which is required by law, was not always respected. This was partly because the administrative authority had accumulated delays in nominating members. Another explanation could be the absence of remuneration for the work and attendance of the members. There was considerable inter-committee variation concerning the way documents were distributed to the members during sessions and the extent of investigator and promoter involvement. Finally, large differences in the number of dossiers handled between committees led us to investigate the fairness of dossier distribution, as the number of dossiers dealt with has consequences, particularly for the finality of the opinions of the committees. CONCLUSIONS: This evaluation of the committees playing a crucial role in protecting participants in research trials provides new information that could be helpful for improving the way in which these committees function in the context of the European Directive. PMID- 15546686 TI - Ciprofloxacin at low levels disrupts colonization resistance of human fecal microflora growing in chemostats. AB - We studied the in vitro effects of a range of ciprofloxacin (CI) concentrations on the human intestinal flora's colonization resistance (CR) to Salmonella kedougou NCTC 12173. Four steady state microbial communities were established in chemostats using inocula from a single pool of human feces. Three chemostats were exposed to CI (0.1, 0.43 and 5 microg/mL, respectively); one served as a no-drug control. The CR of each community was tested by three successive daily challenges of 10(8) S. kedougou, each delivered in a 1 mL bolus. There was no colonization of the no-drug chemostat. Likewise, after exposure to only 0.1 microg/mL CI there was no loss of CR and S. kedougou did not colonize. Conversely, both the 0.43 and the 5 microg/mL-exposed floras suffered a loss of CR and these chemostats were colonized. S. kedougou overgrew faster and reached higher counts in the presence of 0.43 than it did in the presence of 5 microg/mL. One possible explanation is that CI had a dose-dependent effect on both the challenge strain and CR. Thus, at higher levels, even though CR was disrupted by CI, so too was the growth of the challenge strain. Since exposure to CI elicited a dose-dependent reduction in Escherichia coli counts [Reg. Pharmacol. Toxicol. 33 (2001) 276] our new data suggest that E. coli may contribute to the CR against salmonella. We further conclude that, even at fecal levels below those reached during therapy, CI may impact the human gut flora sufficiently to facilitate colonization by S. kedougou. PMID- 15546687 TI - Legal constraints in EU product labelling to mitigate the environmental risk of veterinary medicines at use. AB - This paper summarises what possibilities and obligations are created by the EU Directive 2001/82/EC on the registration of veterinary medicines to mitigate the environmental impact of the use of a veterinary medicinal product. More specifically, an identified environmental risk may be mitigated to an acceptable level by special precautions in the information that accompanies the product in labelling and packaging. These precautions can address the fate of contaminated slurry and treated animals, but are only acceptable under the EU Directive if their effect can be demonstrated using the risk assessment methodology. Next, all possible keepers of the animals or the manure, including third parties, should be addressed, either in the precaution, or in the national regulation that should enforce the precautions. A number of examples illustrate that some precautions used are not quantifiable in the risk methodology, and that others are legally inadequate. To render risk mitigation measures effective, hence suitable for labelling and packaging, it is imperative that the risk assessment methodology is further developed and applied adequately, and that the legality of precautions is established in national regulation, harmonised between Member States. PMID- 15546688 TI - Regulatory immunotoxicology: does the published evidence support mandatory nonclinical immune function screening in drug development? AB - Recent immunotoxicity guidance documents from the EU CHMP and the US FDA apply significantly different weightings to immune function testing; whereas the former mandates (as a starting point) incorporation of immune function tests (IFTs) to screen for immunotoxic potential in sub-chronic rodent toxicity studies, the more cautious 'for cause' FDA approach recommends the use of IFTs only when warranted by evidence obtained from conventional nonclinical and/or clinical studies. Conclusions from detailed evaluations of several key drugs, including salmeterol and some opioids, challenge the notion that data on these examples support the need for IFTs to detect unintended immunosuppression. Given the virtual absence of convincing pharmaceutical examples and the rarity of unintended immunosuppression, routine immune function testing of all new pharmaceuticals is not considered justified. Resources currently being employed in this manner in an attempt to detect a seemingly rare phenomenon would appear to be better applied to the development of reliable predictive assays for drug hypersensitivity, which is known to cause significant patient morbidity. Any moves towards a globally harmonised guideline that recommends the use of concern-based IFTs, need ideally to be accompanied by the establishment of appropriate historical control reference intervals and interpretation criteria to support a reliable weight-of evidence approach to data evaluation. PMID- 15546689 TI - Subacute toxicity evaluation of a new camptothecin anticancer agent CKD-602 administered by intravenous injection to rats. AB - The subacute toxicity of a new camptothecin anticancer agent, CKD-602, was investigated after 4-week repeated intravenous administration of the chemical in Sprague-Dawley rats. The test chemical was administered intravenously to rats at dose levels of 0, 0.003, 0.013, or 0.067 mg/kg/day for males and 0, 0.004, 0.018, or 0.089 mg/kg/day for females. At the end of the treatment period, 10 rats/sex/group were sacrificed. The remaining 5 rats/sex in the vehicle control and high dose groups continued the study without treatment for 2 weeks (recovery period). During the test period, clinical signs, mortality, body weights, food and water consumption, ophthalmoscopy, urinalysis, hematology, serum biochemistry, gross findings, organ weights, and histopathology were examined. In both sexes of the high dose group, an increase in the incidence of abnormal clinical signs and paleness of the eyes, a reduction in the body weight gain, food consumption and urine protein, and an increase in the water consumption were observed. Hematological investigations revealed a decrease in the red blood cells, hemoglobin and hematocrit and an increase in the mean corpuscular volume, mean corpuscular hemoglobin, platelets, and reticulocytes in a dose-dependent manner. Serum total cholesterol and total protein values were lower in females than those of controls, but not in males. An increase in the heart and liver weights and a decrease in the thymus weight were also found. Histopathological alterations included an increase in the incidence of atrophy of the sternal marrow, atrophy, fibrosis and mast cell hyperplasia of the femoral marrow, atrophy of the white pulp and extramedullary hematopoiesis of the spleen, atrophy of the thymus, auricular hypertrophy of the heart, extramedullary hematopoiesis and centriacinar telangiectasis of the liver, follicular degeneration of the ovary, and inflammation of the tail. The major treatment-related effects were not recovered at the end of 2-week recovery period. There were no adverse effects in the low and middle dose groups of both genders. In the present experimental conditions, the target organs were determined to be bone marrow, blood cells, spleen, liver, thymus, and heart. The no-observed-adverse-effect level was considered to be 0.013 mg/kg/day for males and 0.018 mg/kg/day for females. PMID- 15546690 TI - Additional support for derivation of an acute/subchronic reference level for arsenic. PMID- 15546692 TI - Measurement trumps theory. PMID- 15546694 TI - Critical effect of perchlorate on neonates is iodide uptake inhibition. PMID- 15546696 TI - Interspecies differences in susceptibility to perturbation of thyroid hormone homeostasis requires a definition of "sensitivity" that is informative for risk analysis. PMID- 15546698 TI - Reduced temporal lobe volume in early onset conduct disorder. AB - Regional brain volumes derived from magnetic resonance imaging (MRI) scans from 10 youths with early onset conduct disorder and 10 healthy controls matched for age, sex and handedness were compared to determine whether prefrontal or temporal lobe brain volumes differed in the two groups. Right temporal lobe and right temporal gray matter volumes were significantly reduced in subjects with conduct disorder compared with controls. Prefrontal volumes in subjects with conduct disorder were 16% smaller than in controls, but the difference did not reach statistical significance. Early onset conduct disorder without substance abuse comorbidity was also significantly associated with smaller right temporal gray volumes. Further investigation of both the temporal and frontal localizations of the pathophysiology of early onset conduct disorder is warranted in larger samples. PMID- 15546699 TI - A PET provocation study of generalized social phobia. AB - In an investigation of the neural circuits that may mediate the subjective experience of social phobia (SP), six male patients with a primary DSM-IV diagnosis of generalized social phobia watched, in the presence of a group of "communication experts," a videotape of themselves giving an impromptu talk (Exposure condition). In the control Baseline condition, they viewed a videotape of a socially competent stranger giving a talk. Regional cerebral blood flow was measured thrice under each condition. The study revealed significant deactivations from Baseline during Exposure in the right lingual gyrus (BA 18) and in the right medial frontal gyrus (BA 11); significant activations during Exposure were not observed. Deactivation of these regions may reflect a strategy of visual avoidance employed by the patients to dampen their phobic experience. PMID- 15546700 TI - Differences of cerebral activation between superior and inferior learners during motor sequence encoding and retrieval. AB - Cerebral activation during memory encoding and retrieval might depend on subjects' learning capacity, either by corresponding to better performance in superior learners or by reflecting increased effort in inferior learners. To investigate these alternative hypotheses, the study compared cerebral activation during encoding and retrieval of a motor sequence in groups of subjects with superior and inferior learning performances. Ten healthy subjects underwent functional magnetic resonance imaging (fMRI) while performing a motor sequence encoding paradigm (i.e. finger tapping sequence) and a retrieval paradigm (i.e. reproduction of the learned sequence). Subjects were divided into superior and inferior learners according to the correctness of sequence reproduction during retrieval. During encoding, there was strong bilateral activation in the middle frontal gyrus, the supplementary motor area (SMA), the lateral parietal lobe and the cerebellum. During retrieval, again strong activation was found in identical areas of the prefrontal cortex, the parietal lobe and the cerebellum. During encoding, inferior learners showed more left-sided activations in the left middle frontal and inferior parietal gyri. Superior learners showed increased activation in the corresponding right-sided brain areas during encoding as well as during retrieval. Differences of cerebral activations in the prefrontal and parietal cortex during encoding and retrieval were found to be related to retrieval performance, i.e. success and effort. Further intervention studies are needed to assess whether these interindividual differences are the cause or the consequence of differences in memory performance. PMID- 15546701 TI - Comparative study of proton and phosphorus magnetic resonance spectroscopy in schizophrenia at 4 Tesla. AB - This study used high-field magnetic resonance spectroscopy to examine the correlation of 1H and 31P metabolite levels in patients with schizophrenia and normal controls. 1H and 31P in vivo spectra were acquired successively from the left anterior cingulate and left thalamus of nine chronic schizophrenic patients and eight comparable healthy controls. A significant positive correlation between glutamine (Gln) and phosphoethanolamine (PEtn) was found in the left anterior cingulate of patients. In the left thalamus of patients, a significant negative correlation between N-acetylaspartate (NAA) and glycerophosphocholine (GroPCho) was found. No significant correlations were found in controls. The correlation between glutamine and phosphoethanolamine may reflect a link between neurotransmission alterations and membrane phospholipid metabolism alterations. The negative correlation between N-acetylaspartate and glycerophosphocholine may reflect the presence of neurodegeneration. PMID- 15546702 TI - Volume reduction of the amygdala in patients with schizophrenia: a magnetic resonance imaging study. AB - The amygdala is known to be involved in the pathology of schizophrenia. While only a limited number of studies in schizophrenia have measured the amygdala as a single structure. The aim of this study was to examine the hypothesis that patients with schizophrenia would show reduced volumes in the amygdala compared with normal controls. We investigated amygdala volume in 40 patients with schizophrenia (20 males, 20 females) and 40 age- and gender-matched normal controls using three-dimensional magnetic resonance imaging (MRI). Whole volumes of both the amygdala and the temporal lobe were measured on consecutive coronal 1 mm slices. The amygdala volume was significantly smaller in schizophrenia patients than in controls. Considering gender differences, male patients had significantly smaller volumes in the bilateral amygdala than male controls; female patients had a significantly reduced right amygdala compared with female controls. Furthermore, a significant left-smaller-than-right volumetric asymmetry of the amygdala was detected in male patients with schizophrenia. The results may be important for understanding the role of the amygdala in the pathophysiology of schizophrenia and the anatomical substrates of gender difference in the expressions of the illness. PMID- 15546703 TI - Validation of semiautomated methods for quantifying cingulate cortical metrics in schizophrenia. AB - This paper validates semiautomated methods for reconstructing cortical surfaces of the cingulate gyrus from high-resolution magnetic resonance (MR) images. Bayesian segmentation was used to delineate the image voxels into five tissue types: cerebrospinal fluid (CSF), gray matter (GM), white matter (WM), and partial volumes of CSF/GM and GM/WM; the tissues were then recalibrated as CSF, GM, and WM via the Neyman-Pearson Likelihood Ratio Test. To generate cortical surfaces at the interface of GM and WM, the thresholds between the tissue types were first used to reassign partial volume voxels to CSF, GM, and WM with minimum error (that varied from 0.06 to 0.15 for the 10 subjects). Next, topology-correct cortical surfaces were generated and validated with almost all surface vertices lying within one voxel (0.5 mm) of hand contours. Dynamic programming was used to delineate and extract the cingulate gyrus from the cortical surfaces based on its gyral and sulcal boundaries. The intraclass correlation coefficient for surface area obtained by two raters for all 10 surfaces was 0.82. In addition, by repeating the entire procedure three times in one subject, we obtained a coefficient of variation of 0.0438 for surface area. PMID- 15546704 TI - A meta-analysis of functional neuroimaging in obsessive-compulsive disorder. AB - Recent neurobiological models of obsessive-compulsive disorder (OCD) posit that a dysfunction in orbitofrontal-subcortical circuitry underlies the etiology of this disorder. Much of the empirical support for these theories comes from studies using neuroimaging techniques to compare brain activity in OCD patients with that in non-OCD controls. Qualitative reviews of this literature implicate the orbitofrontal cortex, caudate nuclei, and thalamus. In this study, a meta analysis was conducted to summarize the results of studies using positron emission tomography (PET) and single photon emission computed tomography (SPECT) to investigate brain activity in OCD. Results suggest that differences in radiotracer uptake between patients with OCD and healthy controls have been found consistently in the orbital gyrus and the head of the caudate nucleus. No other significant differences were found. The implications of these results for theories regarding the etiology of OCD are discussed. PMID- 15546705 TI - 3 Tesla magnetic resonance imaging of the brain in newborns. AB - While it has been hypothesized that brain development is abnormal in schizophrenia and other neurodevelopmental disorders, there have been few attempts to study very early brain development in children. Twenty unsedated healthy newborns underwent 3 Tesla magnetic resonance imaging (MRI), including diffusion tensor imaging (DTI). The left ventricle was significantly larger than the right; females had significantly larger ventricles than males. Fractional anisotropy (FA) increased significantly with gestational age in the genu and splenium of the corpus callosum. It is feasible to study brain development in unsedated newborns using 3 T MRI. PMID- 15546706 TI - Relationship between performance on the Stroop test and N-acetylaspartate in the medial prefrontal cortex in deficit and nondeficit schizophrenia: preliminary results. AB - The aim of this research was to investigate the relationship between performance on the Stroop test and N-acetylaspartate/creatine assessed using proton magnetic resonance spectroscopy in the medial prefrontal cortex (MPFC) of schizophrenia patients. The Schedule for the Deficit Syndrome was used to subdivide the schizophrenia patients into deficit (n=5) and nondeficit (n=17) subtypes. Twenty one control subjects served as a comparison group. A strong correlation between right-sided N-acetylaspartate/creatine levels and Stroop scores was found in the deficit patients but not in the nondeficit patients and the controls. This result suggests a relationship between a dysfunction of the right medial prefrontal cortex and a deficit in selective attention in schizophrenia patients with the deficit syndrome. PMID- 15546707 TI - Tryptamine-based human beta3-adrenergic receptor agonists. Part 1: SAR studies of the 7-position of the indole ring. AB - A series of tryptamine-based 2-thiophenesulfonamide derivatives were prepared and their agonistic activity for the beta-adrenergic receptors (ARs) was evaluated. Compound 54, containing 7-methanesulfonyloxy tryptamine, was found to be a highly potent beta3-AR agonist (EC50=0.21 nM, IA=97%) with excellent selectivity for the beta3-AR over the beta1- and beta2-ARs (210- and 86-fold, respectively). PMID- 15546708 TI - Tryptamine-based human beta3-adrenergic receptor agonists. Part 2: SAR of the methylene derivatives. AB - A series of tryptamine derivatives with modified sulfonamide were designed, synthesized, and evaluated for their ability to stimulate cAMP accumulation in CHO cells expressing the cloned human beta3-adrenergic receptor (AR). For this series of compounds, our objective was to symmetrize the alpha-position of the tryptamine moiety maintaining its activity and reducing the cost of production. Compound 11h, having m-aminobenzene, exhibited excellent agonistic activity for beta3-AR with excellent subtype selectivity. PMID- 15546709 TI - Synthesis and antitrypanosomal profile of new functionalized 1,3,4-thiadiazole-2 arylhydrazone derivatives, designed as non-mutagenic megazol analogues. AB - In this work we reported the synthesis and the trypanocidal profile of new 1,3,4 thiadiazole-2-arylhydrazone derivatives of nitroimidazole series (4) or phenyl series (5), designed by exploring the molecular hybridization approach between megazol (2) and guanyl hydrazone derivative (3). The evaluation of the activity against bloodstream trypomastigote forms of Trypanosoma cruzi forms lead us to identify a new potent trypamomicide prototype, that is, brazilizone A (4k), which present an IC50/24 h=5.3 microM. PMID- 15546710 TI - Development of a new class of potential antiatherosclerosis agents: NO-donor antioxidants. AB - A new class of NO-donor phenol derivatives is described. The products were obtained by joining appropriate phenols with either nitrooxy or 3 phenylsulfonylfuroxan-4-yloxy moieties. All the compounds proved to inhibit the ferrous salt/ascorbate induced lipidic peroxidation of membrane lipids of rat hepatocytes. They were also capable of dilating rat aorta strips precontracted with phenylephrine. PMID- 15546711 TI - Isosteric N-arylpiperazine replacements in a series of dihydropyridine NPY1 receptor antagonists. AB - 4-Amino-N-arylpiperidines serve as effective bioisosteres for N-arylpiperazines in the series of dihydropyridine NPY1 receptor antagonists. These were prepared by a ZnCl2-mediated reductive amination reaction between elaborated primary amines, 2 or 5, and 4-arylpiperidones. PMID- 15546712 TI - Multidrug resistance reversal activity of permethyl ningalin B amide derivatives. AB - A series of amide derivatives of permethyl ningalin B were prepared and examined as multidrug resistance (MDR) reversal agents illustrating that the C5 carboxylate is widely tolerant of such derivatization. PMID- 15546713 TI - Synthesis and interfacial behavior of sulfur-containing analogs of lung surfactant dipalmitoyl phosphatidylcholine. AB - Synthesis methods and initial surface property characterizations are reported for two sulfur-containing phosphonolipids related structurally to dipalmitoyl phosphatidylcholine (DPPC), the major lung surfactant glycerophospholipid. Sulfur linkages in these compounds affect molecular interactions relative to ester linkages, and are structurally resistant to cleavage by phospholipases. The SO2 linked analog synthesized here had increased adsorption and improved film respreading compared to DPPC, while reaching very low surface tensions (1 N/m) in cycled interfacial films on both the Wilhelmy balance and the pulsating bubble surfactometer. This compound appears to have potential utility as a component in future phospholipase-resistant synthetic exogenous surfactants for treating clinical forms of inflammatory lung injury. PMID- 15546714 TI - Identification of a novel non-carbohydrate molecule that binds to the ribosomal A site RNA. AB - We report the identification of a novel compound that binds to the Escherichia coli 16S ribosomal A-site. Binding by the compound was observed using nuclear magnetic resonance and mass spectrometry techniques. We show that the compound binds in the same position in the A-site RNA as occupied by the aminoglycoside class of antibiotics. PMID- 15546715 TI - Alpha-1-C-alkyl-1-deoxynojirimycin derivatives as potent and selective inhibitors of intestinal isomaltase: remarkable effect of the alkyl chain length on glycosidase inhibitory profile. AB - A series of alpha- and beta-1-C-alkyl-1-deoxynojirimycin derivatives was prepared and evaluated as glycosidase inhibitors. Biological assays showed a marked dependence of the selectivity and potency of the inhibitors upon the position of the alkyl chain (alpha-1-C-, beta-1-C- or N-alkyl derivatives). In addition, the efficiency of alpha-1-C-alkyl-1-deoxynojirimycin derivatives as intestinal isomaltase inhibitors increases with the length of the alkyl chain. The strongest inhibition was found for alpha-1-C -nonyl-1-deoxynojirimycin with an IC50=3.5 nM (25x more potent inhibitor than the shorter chain homologue carrying a C8 chain). These results demonstrate that subtle changes in the aglycon fragment may result in remarkable enzyme specificity. PMID- 15546716 TI - Isoxazole-3-hydroxamic acid derivatives as peptide deformylase inhibitors and potential antibacterial agents. AB - A series of isoxazole-3-hydroxamic acid derivatives has been identified as a new class of small, nonpeptidic inhibitors of peptide deformylase (PDF). The synthesis, enzyme inhibition and preliminary investigation of the binding mode of this potential antibacterial compounds are reported. PMID- 15546717 TI - Carbonic anhydrase inhibitors. Inhibition of the prokariotic beta and gamma-class enzymes from Archaea with sulfonamides. AB - A detailed inhibition study of carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the beta- and gamma-families from Archaea with sulfonamides has been performed. Compounds included in this study were the clinically used sulfonamide CA inhibitors, such as acetazolamide, methazolamide, ethoxzolamide, topiramate, valdecoxib, celecoxib, dorzolamide, sulfanilamide, dichlorophanamide, as well as sulfanilamide analogs, halogenated sulfanilamides, and some 1,3 benzenedisulfonamide derivatives. The two gamma-CAs from Methanosarcina thermophila (Zn-Cam and Co-Cam) showed very different inhibitory properties with these compounds, as compared to the alpha-CA isozymes hCA I, II, and IX, and the beta-CA from Methanobacterium thermoautotrophicum (Cab). The best Zn-Cam inhibitors were sulfamic acid and acetazolamide, with inhibition constants in the range of 63-96 nM, whereas other investigated aromatic/heterocylic sulfonamides showed a rather levelled behavior, with KIs in the range of 0.12-1.70 microM. The best Co-Cam inhibitors were topiramate and p-aminoethyl-benzenesulfonamide, with KIs in the range of 0.12-0.13 microM, whereas the worst one was homosulfanilamide (KI of 8.50 microM). In the case of Cab, the inhibitory power of these compounds varied to a much larger extent, with sulfamic acid and sulfamide showing millimolar affinities (KIs in the range of 44-103 mM), whereas the best inhibitor was ethoxzolamide, with a KI of 5.35 microM. Most of these sulfonamides showed inhibition constants in the range of 12-100 microM against Cab. Thus, the three CA families investigated up to now possess a very diverse affinity for sulfonamides, the inhibitors with important medicinal, and environmental applications. PMID- 15546718 TI - Synthesis of 8-thiabicyclo[3.2.1]oct-2-enes and their binding affinity for the dopamine and serotonin transporters. AB - The reinforcing and stimulant properties of cocaine have been primarily associated with its propensity to bind to monoamine transport systems, in particular the dopamine transporter. Inhibition of the dopamine transporter then leads to an increase of synaptic dopamine with substantial pharmacological consequences. The search for medications for cocaine abuse has had a particular focus on tropane analogs of cocaine, and the interchange of nitrogen for oxygen in this class has led to potent and selective inhibitors of monoamine transport. Herein we report that 8-thiatrop-2-enes are highly potent and quite selective inhibitors of the dopamine transporter. The 3,4-dichlorophenyl-8 thiabicyclo[3.2.1]oct-2-ene (4f) is particularly potent (IC50=4.5 nM) and selective (800-fold) with respect to inhibition of the serotonin transporter. PMID- 15546719 TI - Design, synthesis and evaluation of bicyclic benzamides as novel 5-HT1F receptor agonists. AB - Several fused bicyclic systems have been investigated to serve as the core structure of potent and selective 5-HT1F receptor agonists. Replacement of the indole nucleus in 2 with indazole and 'inverted' indazole provided more potent and selective 5-HT1F receptor ligands. Indoline and 1,2-benzisoxazole systems also provided potent 5-HT1F receptor agonists, and the 5-HT1A receptor selectivity of the indoline- and 1,2-benzisoxazole-based 5-HT1F receptor agonists could be improved with modification of the benzoyl moiety of the benzamides. Through these studies, we found that the inherent geometries of the templates, not the nature of hybridization of the linking atom, were important for the 5 HT1F receptor recognition. PMID- 15546721 TI - Tocopherol long chain fatty alcohols decrease the production of TNF-alpha and NO radicals by activated microglial cells. AB - The synthesis of a series of Tocopherol long chain Fatty Alcohols (TFA) and their biological activities on the modulation of microglial activation are described. Specifically, the 2-(12-hydroxy-dodecyl)-2,5,7,8-tetramethyl-chroman-6-ol, the TFA bearing 12 carbon atoms on the side chain (n=12), shows the most potent inhibition of secretion on nitric oxide (NO) and tumour necrosis factor-alpha (TNF-alpha) by lipopolysaccharide (LPS)-activated microglia. PMID- 15546720 TI - CVT-4325: a potent fatty acid oxidation inhibitor with favorable oral bioavailability. AB - New inhibitors of palmitoyl-CoA oxidation are based on the introduction of nitrogen heterocycles in the 'Western Portion' of the molecule. SAR studies led to the discovery of CVT-4325 (shown), a potent FOXi (IC50=380 nM rat mitochondria) with favorable PK properties (F=93%, t(1/2)=13.6h, dog). PMID- 15546722 TI - Synthesis, (1-->3)-beta-D-glucanase-binding ability and phytoalexin-elicitor activity of (R)-2,3-epoxypropyl (1-->3)-beta-D-pentaglucoside. AB - The (1-->3)-beta-D-pentaglucoside was synthesized as its (R)-2,3-epoxypropyl glycoside via 2+3 strategy. The disaccharide donor 8 was obtained by 3-selective coupling of 2 with 4, followed by deallylation, and trichloroacetimidation. Meanwhile, the trisaccharide acceptor 12 was prepared by coupling of 10 with 4, followed by deacetylation. Condensation of 8 with 12, followed by epoxidation, and deprotection, gave the target pentaoside. The results of these bioassays demonstrated that the (1-->3)-beta-D-glucanase was obviously inactivated by 15 with k(app)=3.79 x 10(-4) min(-1). At the same time, we found that the 15 was more active as compared to the laminaripentaose in eliciting phytoalexin accumulation in tobacco cotyledon tissue, and it could be kept longer time than laminaripentaose, which indicated it is much more stable than laminaripentaose. PMID- 15546723 TI - Fluorescence quenching of parallel-stranded DNA bound ethidium bromide: the effect of 7-deaza-2'-deoxyisoguanosine and 7-halogenated derivatives. AB - Parallel stranded (ps) duplexes were constructed by incorporating 7-deaza-2' deoxyisoguanosine (1a) or its 7-halogenated analogs 1b,c in place of 2' deoxyisoguanosine. UV and Tm analyses prove the high affinity of ethidium bromide (EB) to these modified duplexes. Steady-state fluorescence measurement shows that the fluorescence is quenched when EB is bound to ps duplexes containing compounds 1a-c. The quenching effect depends on the 7-substituent of the nucleobase. PMID- 15546724 TI - Design, synthesis and anti-microbial activity of 1H-pyrazole carboxylates. AB - In a SAR study, we have synthesized a few 1H-pyrazole carboxylate related microbicides using Vilsmeier reagent. The anti-microbial screening results of 1H pyrazole-3-carboxylate are reported here for the first time. The effect of 1H pyrazole carboxylates on the mycelial growth of plant pathogenic fungi is revealed. The first X-ray structure in the family of microbicidal 1H-pyrazole-4 carboxylates is presented. PMID- 15546725 TI - Antiviral compounds from traditional Chinese medicines Galla Chinese as inhibitors of HCV NS3 protease. AB - Under the guidance of bioassay, the EtOAc extract fraction of the Traditional Chinese Medicine (TCM) Galla Chinese was found to be efficient in inhibiting the NS3 protease of HCV and purified the fraction to get three polyphenol compounds 1,2,6-tri-O-galloyl-beta-D-glucose (1), 1,2,3,6-tetra-O-galloyl-beta-D-glucose (2), and 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (3), which were identified as inhibitors of Hepatitis C Virus (HCV) NS3 protease. Compounds 1, 2, and 3 inhibited HCV NS3 protease with IC50 of 1.89, 0.75, and 1.60 microM, respectively. PMID- 15546726 TI - Development of an efficient and selective radioligand for bradykinin B1 receptor occupancy studies. AB - We have developed an efficient and selective radioligand, the [35S]-radiolabeled dihydroquinoxalinone derivative, 4, for an ex vivo receptor occupancy assay in transgenic rats over-expressing the human bradykinin B1 receptor. PMID- 15546727 TI - 3-(2-Methoxytetrahydrofuran-2-yl)pyrazoles: a novel class of potent, selective cyclooxygenase-2 (COX-2) inhibitors. AB - A series of 3-(2-methoxytetrahydrofuran-2-yl)pyrazoles (4-10) was synthesized. The compounds were evaluated for their ability to inhibit cyclooxygenase-1 (COX 1) and cyclooxygenase-2 (COX-2) activity in human whole blood (HWB). The compound, 5-(4-methanesulfonylphenyl)-3-(2-methoxytetrahydrofuran-2-yl)-1-p-tolyl 1H-pyrazole 5 showed potent and selective COX-2 inhibition (IC50 for COX-1: >100 microM and COX-2: 1.2 microM). PMID- 15546728 TI - Design of bivalent ligands using hydrogen bond linkers: synthesis and evaluation of inhibitors for human beta-tryptase. AB - We exploit the concept of using hydrogen bonds to link multiple ligands for maintaining simultaneous interactions with polyvalent binding sites. This approach is demonstrated by the syntheses and evaluation of pseudo-bivalent ligands as potent inhibitors of human beta-tryptase. PMID- 15546729 TI - Synthesis and anticonvulsant activity of new 2-substituted-5- [2-(2 fluorophenoxy)phenyl]-1,3,4-oxadiazoles and 1,2,4-triazoles. AB - A series of new 2-substituted-5-[2-(2-fluorophenoxy)phenyl]-1,3,4-oxadiazoles has been synthesized and screened for their anticonvulsant activities. Compound 3 shows considerable anticonvulsant activity both in PTZ and MES models. It seems that this effect is mediated by benzodiazepine receptors and other unknown mechanism, respectively. PMID- 15546730 TI - Discovery of novel 2-(aminoheteroaryl)-thiazole-5-carboxamides as potent and orally active Src-family kinase p56(Lck) inhibitors. AB - A series of substituted 2-(aminoheteroaryl)-thiazole-5-carboxamide analogs have been synthesized as novel, potent inhibitors of the Src-family kinase p56Lck. Among them, compound 2 displayed superior in vitro potency and excellent in vivo efficacy. PMID- 15546731 TI - Improving the intein-mediated, site-specific protein biotinylation strategies both in vitro and in vivo. AB - One of the critical issues in the generation of a protein microarray lies in the choice of immobilization strategies, which ensure proteins are adhered to the glass surface while properly retaining their native biological activities. We previously developed intein-mediated strategies for protein biotinylation and site-specific protein microarray generation. Herein, we report new findings of these strategies, which improve the biotinylation efficiency of proteins by up to 10-folds. PMID- 15546732 TI - Structure-based virtual screening and biological evaluation of potent and selective ADAM12 inhibitors. AB - We describe a series of potent and selective inhibitors of ADAM12 that were discovered using computational screening of a focused virtual library. The initial structure-based virtual screening selected 64 compounds from a 3D database of 67,062 molecules. Being evaluated by a cell-based ADAM12 activity assay, compounds 5, 11, 14, 16 were further identified as the potent and selective inhibitors of ADAM12 with low nanomolar IC50 values. The mechanism underlying the potency and selectivity of a representative compound, 5, was investigated through molecular docking studies. PMID- 15546733 TI - Novel diarylsulfonylurea derivatives as potent antimitotic agents. AB - A novel series of diarylsulfonylurea derivatives were synthesized and evaluated for interaction with tubulin and for cytotoxicity against human cancer cell lines. These derivatives demonstrated good inhibitory activity against tubulin polymerization, which was well correlated with promising antiproliferative activity as well as G2/M phase cell cycle arrest. Furthermore, several compounds were also efficacious against multidrug-resistant cancer cells, which are resistant to many other known microtubule inhibitors. PMID- 15546734 TI - Alumina supported potassium permanganate: an efficient reagent for chemoselective dehydrogenation of 2-imidazolines under mild conditions. AB - Potassium permanganate supported on alumina was found to be an efficient reagent system for dehydrogenation of 2-imidazolines to imidazoles under mild conditions at room temperature. Selective oxidation of 2-alkylimidazolines in the presence of 2-arylimidazolines was achieved using this reagent system. 2-Imidazolines were also selectively converted to their corresponding imidazoles in the presence of other oxidizable functional groups such as sulfide, ether, aldehyde, acetal and THP ether. The oxidation procedure described here is easy to carry out and does not require strict reaction conditions. PMID- 15546735 TI - Identification of a highly potent and selective DNA-dependent protein kinase (DNA PK) inhibitor (NU7441) by screening of chromenone libraries. AB - A solution-phase multiple-parallel synthesis approach was employed for the preparation of 6-, 7- and 8-aryl-substituted chromenone libraries, which were screened as inhibitors of the DNA repair enzyme DNA-dependent protein kinase (DNA PK). These studies resulted in the identification of 8-dibenzothiophen-4-yl-2 morpholin-4-yl-chromen-4-one (NU7441) as a highly potent and selective DNA-PK inhibitor (IC50=14 nM), exhibiting ATP-competitive inhibition kinetics. PMID- 15546736 TI - QSAR modelling of HIV-1 reverse transcriptase inhibition by benzoxazinones using a combination of P_VSA and pharmacophore feature descriptors. AB - In pursuit of better anti-HIV drugs, a quantitative structure-activity relationship analysis using a novel set of 2D descriptors was performed on a series of HIV-1 reverse transcriptase inhibitory benzoxazinones. The QSAR models derived from the above mentioned descriptors were found to be statistically significant and exhibited superior predictive power. The results of the study justify the application of the descriptors for exploring the binding mode of the benzoxazinones to the enzyme. PMID- 15546737 TI - Aminoimidazo[1,2-a]pyridines as a new structural class of cyclin-dependent kinase inhibitors. Part 1: Design, synthesis, and biological evaluation. AB - We have identified a novel structural class of protein serine/threonine kinase inhibitors comprised of an aminoimidazo[1,2-a]pyridine nucleus. Compounds from this family are shown to potently inhibit cyclin-dependent kinases by competing with ATP for binding to a catalytic subunit of the protein. Structure-based design approach was used to direct this chemical scaffold toward generating potent and selective CDK2 inhibitors. The discovery of this new class of ATP-site directed protein kinase inhibitors, aminoimidazo[1,2-a]pyridines, provides the basis of new medicinal chemistry tool in search for an effective treatment of cancer and other diseases that involve protein kinase signaling pathways. PMID- 15546738 TI - A comparative binding study of modified bovine immunodeficiency virus TAR RNA against its TAT peptide. AB - Besides generating novel binding peptides or small molecules to their RNA target, successful design of chemically modified RNA constructs capable of tighter binding with their binding peptides is also of significant importance. Herein, the synthesis and binding studies of a series of both wt and mutant bovine immunodeficiency virus (BIV) TAR RNA constructs against its Tat peptide are reported. Understanding the requirements that enable RNA construct binding properties, especially at the hairpin loop or internal bulge, would afford potential therapeutic approaches to control the BIV life cycle. PMID- 15546739 TI - The synthesis and evaluation of [2.2.1]-bicycloazahydantoins as androgen receptor antagonists. AB - A novel series of [2.2.1]-azahydantoins has been designed and synthesized in an enantiospecific manner. The ability of these compounds to act as antagonists to the androgen receptor was investigated and several were found to have potent activity in vitro. PMID- 15546740 TI - Conversion of human-selective PPARalpha agonists to human/mouse dual agonists: a molecular modeling analysis. AB - To understand the species selectivity in a series of alpha-methyl-alpha-phenoxy carboxylic acid PPARalpha/gamma dual agonists (1-11), structure-based molecular modeling was carried out in the ligand binding pockets of both human and mouse PPARalpha. This study suggested that interaction of both 4-phenoxy and phenyloxazole substituents of these ligands with F272 and M279 in mouse PPARalpha leads to the species-specific divergence in ligand binding. Insights obtained in the molecular modeling studies of these key interactions resulted in the ability to convert a human-selective PPARalpha agonist to a human and mouse dual agonist within the same platform. PMID- 15546741 TI - 9-cis-retinoic acid analogues with bulky hydrophobic rings: new RXR-selective agonists. AB - Stille cross-coupling of aryltriflates 10 and dienylstannane 11, oxidation and Horner-Wadsworth-Emmons reaction afforded stereoselectively retinoates 15. Saponification provided the carboxylic acids 8a and 8b, retinoids that incorporate a bulky hydrophobic ring while preserving the 9-cis-geometry of the parent system. In contrast to the pan-RAR/RXR agonistic profile of the lower homologue of 8a, compound 7 (LG100567), retinoids 8 showed selective binding and transactivation of RXR, devoid of significant RAR activation. In PLB985 leukemia cells that require RXR agonists for differentiation compounds 8 induced maturation in the presence of the RAR-selective pan-agonist TTNPB; this effect was blocked by an RXR-selective antagonist. PMID- 15546742 TI - Evans Blue is an inhibitor of nuclear factor-kappa B (NF-kappaB)-DNA binding. AB - Nuclear factor-kappa B (NF-kappaB) is an important transcription factor, involved in many immune and inflammatory responses. It is critical in HIV gene expression as it has kappa B binding sites in the HIV-1 long-terminal repeat. Hence, targeting NF-kappaB to prevent its DNA binding holds a significant therapeutic potential. In this context, we report Evans Blue as a novel inhibitor of NF kappaB-DNA binding. Evans Blue was found to be inhibiting DNA binding of NF kappaB at a low concentration of 100 microM. Further, molecular modeling studies using docking and generation of electrostatic potential maps predicted a possible binding mode of EB to the DNA binding region of NF-kappaB, consistent with the experimental activity. PMID- 15546743 TI - Chimeric cyclodepsipeptides as mimetics for the anthelmintic PF1022A. AB - In the anthelmintic cyclooctadepsipeptide PF1022A (1) didepsipeptide units have been exchanged for the beta-turn mimetics (D)-Pro-(L)-Pro and BTD (7) in order to elucidate the functional role of the depsipeptide backbone. Compounds 12 and 14 are the first PF1022A analogues in which a substantial part of the PF1022A backbone has been replaced with an improvement of anthelmintical activity. Preliminary structure-activity relationships suggest a symmetric conformation to be the biological active one. PMID- 15546744 TI - Random coil phosphorus chemical shift of deoxyribonucleic acids. AB - Random coil phosphorus chemical shift has been studied using 16 17-nucleotide DNA sequences. Due to the presence of residual base stacking in these sequences, the temperature and sequence effects were investigated at 50 and 55 degrees C. The phosphorus chemical shifts of random coil DNA sequences have been found to be independent of temperature. Sequence effect analysis shows that the phosphorus chemical shift of a nucleotide in a random coil DNA sequence depends on both its 5'- and 3'-nearest neighbors. A trimer model has been used to establish the random coil 31P chemical shift prediction protocol which shows an accuracy of 0.02 ppm. PMID- 15546745 TI - Extending the limits of the selective 1D NOESY experiment with an improved selective TOCSY edited preparation function. AB - Compared to its 2D counterpart, the selective 1D NOESY experiment offers greatly simplified spectral interpretation and is invaluable to the structure elucidation of small-to-medium sized molecules, although its application is limited to well resolved resonances only. The doubly selective 1D TOCSY-NOESY experiment allows the 1D NOESY experiment to be extended to resonances within overlapped spectral regions. However, existing methods do not address the critical issue of zero quantum interference, which leads to severe anti-phase distortions to the line shape of scalar coupled spins and often complicates the identification of weak NOE enhancements. In this communication, we describe an improved selective TOCSY edited preparation (STEP) function and its application to the selective 1D NOESY experiment. The STEP function incorporates a novel zero-quantum filter introduced by Thrippleton and Keeler [Angew. Chem. Int. Ed. 42 (2003) 3938], which permits essentially complete suppression of zero-quantum coherence in a single scan. Residual anti-phase distortions due to spin-state mixing are removed using the double difference methodology reported by Shaka et al. [45th Experimental NMR Conference, Pacific Grove, USA, 2004]. The combined use of these techniques ensures that the final spectra are free of distortions, which is crucial to the reliable detection of weak NOE enhancements. Although employed as an additional preparation period in the example demonstrated here, the STEP function affords a general editing tool for spectral simplification and can be applied to a range of experiments. PMID- 15546746 TI - Phase modulation in dipolar-coupled A2 spin systems: effect of maximum state mixing in 1H NMR in vivo. AB - Coupling constants of nuclear spin systems can be determined from phase modulation of multiplet resonances. Strongly coupled systems such as citrate in prostatic tissue exhibit a more complex modulation than AX connectivities, because of substantial mixing of quantum states. An extreme limit is the coupling of n isochronous spins (An system). It is observable only for directly connected spins like the methylene protons of creatine and phosphocreatine which experience residual dipolar coupling in intact muscle tissue in vivo. We will demonstrate that phase modulation of this "pseudo-strong" system is quite simple compared to those of AB systems. Theory predicts that the spin-echo experiment yields conditions as in the case of weak interactions, in particular, the phase modulation depends linearly on the line splitting and the echo time. PMID- 15546747 TI - Molecular dynamics and information on possible sites of interaction of intramyocellular metabolites in vivo from resolved dipolar couplings in localized 1H NMR spectra. AB - Proton NMR resonances of the endogenous metabolites creatine and phosphocreatine ((P)Cr), taurine (Tau), and carnosine (Cs, beta-alanyl-L-histidine) were studied with regard to residual dipolar couplings and molecular mobility. We present an analysis of the direct 1H-1H interaction that provides information on motional reorientation of subgroups in these molecules in vivo. For this purpose, localized 1H NMR experiments were performed on m. gastrocnemius of healthy volunteers using a 1.5-T clinical whole-body MR scanner. We evaluated the observable dipolar coupling strength SD0 (S=order parameter) of the (P)Cr-methyl triplet and the Tau-methylene doublet by means of the apparent line splitting. These were compared to the dipolar coupling strength of the (P)Cr-methylene doublet. In contrast to the aliphatic protons of (P)Cr and Tau, the aromatic H2 (delta=8 ppm) and H4 (delta=7 ppm) protons of the imidazole ring of Cs exhibit second-order spectra at 1.5 T. This effect is the consequence of incomplete transition from Zeeman to Paschen-Back regime and allows a determination of SD0 from H2 and H4 of Cs as an alternative to evaluating the multiplet splitting which can be measured directly in high-resolution 1H NMR spectra. Experimental data showed striking differences in the mobility of the metabolites when the dipolar coupling constant D0 (calculated with the internuclear distance known from molecular geometry in the case of complete absence of molecular dynamics and motion) is used for comparison. The aliphatic signals involve very small order parameters S approximately (1.4-3) x 10(-4) indicating rapid reorientation of the corresponding subgroups in these metabolites. In contrast, analysis of the Cs resonances yielded S approximately (113-137) x 10(-4). Thus, the immobilization of the Cs imidazole ring owing to an anisotropic cellular substructure in human m. gastrocnemius is much more effective than for (P)Cr and Tau subgroups. Furthermore, 1H NMR experiments on aqueous model solutions of histidine and N acetyl-L-aspartate (NAA) enabled the assignment of an additional signal component at delta=8 ppm of Cs in vivo to the amide group at the peptide bond. The visibility of this proton could result from hydrogen bonding which would agree with the anticipated stronger motional restriction of Cs. Referring to the observation that all dipolar-coupled multiplets resolved in localized in vivo 1H NMR spectra of human m. gastrocnemius collapse simultaneously when the fibre structure is tilted towards the magic angle (theta; approximately 55 degrees), a common model for molecular confinement in muscle tissue is proposed on the basis of an interaction of the studied metabolites with myocellular membrane phospholipids. PMID- 15546748 TI - An NMR study of the origin of dioxygen-induced spin-lattice relaxation enhancement and chemical shift perturbation. AB - Due to its depth-dependent solubility, oxygen exerts paramagnetic effects which become progressively greater toward the hydrophobic interior of micelles, and lipid bilayer membranes. This paramagnetic gradient, which is manifested as contact shift perturbations (19F and 13C NMR) and spin-lattice relaxation enhancement (19F and 1H NMR), has been shown to be useful for precisely determining immersion depth, membrane protein secondary structure, and overall topology of membrane proteins. We have investigated the influence of oxygen on 19F and 13C NMR spectra and spin-lattice relaxation rates of a semiperfluorinated detergent, (8,8,8)-trifluoro (3,3,4,4,5,5,6,6,7,7)-difluoro octylmaltoside (TFOM) in a model membrane system, to determine the dominant paramagnetic spin-lattice relaxation and shift-perturbation mechanism. Based on the ratio of paramagnetic spin-lattice relaxation rates of 19F and directly bonded 13C nuclei, we conclude that the dominant relaxation mechanism must be dipolar. Furthermore, the temperature dependence of oxygen-induced chemical shift perturbations in 9F NMR spectra suggests a contact interaction is the dominant shift mechanism. The respective hyperfine coupling constants for 19F and 13C nuclei can then be estimated from the contact shifts <(deltav/v0)19F> and <(deltav/v0)13C>, allowing us to estimate the relative contribution of scalar and dipolar relaxation to 19F and 13C nuclei. We conclude that the contribution to spin-lattice relaxation from the oxygen induced paramagnetic scalar mechanism is negligible. PMID- 15546749 TI - Modulation of the distance dependence of paramagnetic relaxation enhancements by CSA x DSA cross-correlation. AB - Paramagnetic metal ions with fast-relaxing electronic spin and anisotropic susceptibility tensor provide a rich source of structural information that can be derived from pseudo-contact shifts, residual dipolar couplings, dipole-dipole Curie spin cross-correlation, and paramagnetic relaxation enhancements. The present study draws attention to a cross-correlation effect between nuclear relaxation due to anisotropic chemical shielding (CSA) and due to the anisotropic dipolar shielding (DSA) caused by the electronic Curie spin. This CSA x DSA cross correlation contribution seems to have been overlooked in previous interpretations of paramagnetic relaxation enhancements. It is shown to be sufficiently large to compromise the 1/r6 distance dependence usually assumed. The effect cannot experimentally be separated from auto-correlated DSA relaxation. It can increase or decrease the observed paramagnetic relaxation enhancement. Under certain conditions, the effect can dominate the entire paramagnetic relaxation, resulting in nuclear resonances narrower than in the absence of the paramagnetic center. CSAxDSA cross-correlation becomes important when paramagnetic relaxation is predominantly due to the Curie rather than the Solomon mechanism. Therefore the effect is most pronounced for relaxation by metal ions with large magnetic susceptibility and fast-relaxing electron spin. It most strongly affects paramagnetic enhancements of transverse relaxation in macromolecules and of longitudinal relaxation in small molecules. PMID- 15546750 TI - Separation and characterization of different signals from intermolecular three spin orders in solution NMR. AB - In this paper, signals originating from a pure specific coherence of intermolecular three-spin orders were separated and characterized experimentally in highly polarized two-component spin systems. A modified CRAZED sequence with selective radio-frequency excitation was designed to separate the small signals from the strong conventional single-spin single-quantum signals. General theoretical expressions of the pulse sequence with arbitrary flip angle pulses were derived using dipolar field treatment. The expressions were used to predict the relaxation and diffusion properties and optimal experimental parameters such as flip angles. For the first time, relaxation and diffusion properties of pure intermolecular single-quantum, double-quantum, and triple-quantum coherences of three-spin orders were characterized and analyzed in one-dimensional experiments. All experimental observations are in excellent agreement with the theoretical predictions. The theoretical results show that the quantum-mechanical treatment leads to exactly the same predictions as the dipolar field treatment. The quantitative study of intermolecular multiple-quantum coherences of three-spin orders presented herein provides a better understanding of their mechanisms. PMID- 15546751 TI - Magnetic relaxation dispersion probe. AB - The magnetic field dependence of nuclear spin-lattice relaxation rates provides a powerful approach to characterizing intra and intermolecular dynamics. NMR spectrometers that provide extensive magnetic relaxation dispersion profiles may switch magnetic field strengths rapidly by either moving the sample or by changing the current in an electromagnet. If the sample is moved, the polarization and detection fields may be very high, which provides both high sensitivity and resolution. This report summarizes the design of a pneumatic sample transport system for glass sample containers that may be used in either a dual magnet spectrometer or in a single magnet system that exploits the fringe field as the secondary magnetic field. PMID- 15546752 TI - Weak alignment of membrane proteins in stressed polyacrylamide gels. AB - Residual dipolar couplings are important as angular constraints for the structure determination of membrane proteins in micelles. Strained polyacrylamide gels are one of the few available mechanisms available for inducing the requisite weak alignment for these samples. However, their use is frequently limited by the ability to incorporate proteins and buffer solutions into the gel matrix. The implementation of several methods of incorporating membrane proteins into gels are described. Conditions for copolymerizing the protein in the absence of a change in pH are detailed. Electrophoresis is also shown to be a useful method to incorporate proteins. Weak alignment of the protein-micelle complex in the gel matrix is subsequently achieved using either vertical or radial compression. The magnitude of alignment can be controlled by altering the gel concentration, the acrylamide/bisacrylamide ratio, and the compression ratio. The alignment tensor can be altered relative to uncharged polyacrylamide gels by copolymerizing samples with acrylamide/acrylic acid to incorporate negative charges in the strained polyacrylamide gel to provide an alternate orientation. PMID- 15546753 TI - Sensitivity-enhanced double-TROSY experiment for simultaneous measurement of one bond 15N-1H, 15N-13C' and two-bond 1H-13C' couplings. AB - A recently published experiment for the measurement of 1JHN, 1JNC', and 2JHC' coupling constants [J. Am. Chem. Soc. 125 (2003) 11504] was modified to yield a double-TROSY experiment which selects 1 of the 16 multiplet components from a 15N HSQC spectrum recorded of a uniformly 15N/13C-labelled protein. Subspectra containing any 1 of the 16 multiplet components can be generated allowing accurate coupling constant measurements. The experiment is sensitivity enhanced, turning all magnetization components precessing during the evolution time into observable magnetization during the detection time. The experiment is discussed with regard to the previously published alpha/beta-filtered HN(alpha/beta-NC'-J) experiment [J. Magn. Reson. 140 (1999), 32] which measures the same coupling constants. PMID- 15546754 TI - Covariance NMR spectroscopy by singular value decomposition. AB - Covariance NMR is demonstrated for homonuclear 2D NMR data collected using the hypercomplex and TPPI methods. Absorption mode 2D spectra are obtained by application of the square-root operation to the covariance matrices. The resulting spectra closely resemble the 2D Fourier transformation spectra, except that they are fully symmetric with the spectral resolution along both dimensions determined by the favorable resolution achievable along omega2. An efficient method is introduced for the calculation of the square root of the covariance spectrum by applying a singular value decomposition (SVD) directly to the mixed time-frequency domain data matrix. Applications are shown for 2D NOESY and 2QF COSY data sets and computational benchmarks are given for data matrix dimensions typically encountered in practice. The SVD implementation makes covariance NMR amenable to routine applications. PMID- 15546755 TI - Molecular-orientation analysis based on alignment-induced TROSY chemical shift changes. AB - We present a new NMR technique for determining the alignment tensor of a weakly aligned protein using only alignment-induced 15N transverse relaxation optimized spectroscopy (TROSY) chemical shift changes. Alignment-induced TROSY chemical shift changes reflect the combined contributions from two different anisotropic spin interactions including the residual dipolar couplings (RDCs) and the residual chemical shift anisotropy effects (RCSAs). We show here that these two residual anisotropic spin interactions' values, encoded in the TROSY chemical shift changes, can be used to determine a weakly aligned protein's alignment tensor. To prove the significance of this method, we show that our TROSY-based analysis gives the consistent alignment angles with those determined using RDCs for 15N-labeled ubiquitin (8.6 kDa) in an aligned medium, within an uncertainty range estimated by considering experimental and structural noises, being 5 degrees at most. Because our approach requires a pre-determined 15N CSA tensor value, we also estimated the uncertainties associated with the resultant alignment tensor values caused by variation in 15N CSA tensors. In spite of the significant variations in literature-reported 15N CSA tensors, they gave consistent orientation angles within an uncertainty range. These results ensure that our TROSY-based approach is a useful alternative to the RDC-based method to determine the alignment angles especially for large proteins in a weakly aligned state. PMID- 15546756 TI - Molecular dynamics of half-integer quadrupolar nuclei studied by QCPMG solid state NMR experiments on static and rotating samples. Theory and simulations. AB - Simulations of QCPMG NMR type experiments have been used to explore dynamic processes of half-integer quadrupolar nuclei in solids. By setting up a theoretical approach that is well suited for efficient numerical simulations the QCPMG type experiments have been analyzed regarding the effect of the magnitude of the EFG- and CSA-tensors, the spin-quantum number, different dynamical processes and MAS. Compared to the QE experiment the QCPMG experiment offers not only intensity gain by an order of magnitude and changes in overall lineshape as a function of the kinetic rate constant but the lineshape of the individual spin echo sidebands is also very sensitive towards dynamics. Hereby a visual identification of the dynamics is obtained. In common for all the simulations the spin-echo sidebands are narrow in the slow (k< or =10(2) Hz) and the fast (k> or =10(7) Hz) dynamic regime whereas they are broadened in the intermediate regime 10(3)< or =k< or =10(7) Hz. The maximum intensity of the spin-echo sidebands for two-site jumps is highly dependent on the type of anisotropic interactions involved and the type of QCPMG experiment. Hence, in the fast limit the maximum intensity was 140% of the initial intensity when significant CSA was present or under the QCPMG-MAS experiment compared to 89 or 71% for the static experiment influenced by the quadrupolar interaction only. For 3-, 4-, and 6-site jumps the maximum intensity in the fast limit reached up to 339% of the intensity in the static limit. PMID- 15546757 TI - Exact half pulse synthesis via the inverse scattering transform. AB - In a paper of Nielson et al. it is shown, using the linear approximation, that it might be possible to create a pair of RF-pulses, which, after summation of the unrephased signals achieve a specified transverse magnetization. Such pulses, designed using the linear approximation, show rather poor slice selectivity. Using the inverse scattering transform formalism we give an algorithm to exactly achieve a specified "summed" transverse magnetization profile. Indeed for a constant phase transverse profile, our algorithm produces infinitely many solutions. PMID- 15546758 TI - Cross polarization, radio frequency field homogeneity, and circuit balancing in high field solid state NMR probes. AB - Homogeneous radio frequency (RF) fields are important for sensitivity and efficiency of magnetization transfer in solid state NMR experiments. If the fields are inhomogeneous the cross polarization (CP) experiment transfers magnetization in only a thin slice of sample rather than throughout the entire volume. Asymmetric patterns have been observed in plots of the CP signal versus RF field mismatch for an 800 MHz solid-state NMR probe where each channel is resonated in a single-ended mode. A simple model of CP shows these patterns can be reproduced if the RF fields for the two nuclei are centered at different places in the coil. Experimental measurements using B1 field imaging, nutation arrays on extremely short NMR samples, and de-tuning experiments involving disks of copper incrementally moved through the coil support this model of spatially offset RF fields. We have found that resonating the high frequency channel in a double-ended or "balanced" mode can alleviate this field offset problem, and have implemented this in a three-channel solid state NMR probe of our own design. PMID- 15546759 TI - Analysis of the magic asymmetric gradient stimulated echo sequence with shaped gradients. AB - Recently a 13-interval magic asymmetrical gradient stimulated echo (MAGSTE) sequence has been proposed for accurate displacement measurements in the presence of spatially varying background gradients. In this paper, the commonly used trapezoidal and sine shaped gradients are studied for the MAGSTE sequence, and the magic asymmetrical gradient ratio and b-factor are provided. The derivation enables the MAGSTE sequence to be implemented on systems with non-negligible gradient rise times. PMID- 15546760 TI - Product operator analysis of the influence of chemical exchange on relaxation rates. AB - Measurements of chemical-exchange processes by NMR are widely used to obtain valuable information about molecular dynamics and structure. Here, a computational method is introduced to assess the influence of chemical exchange on spin relaxation rates. The method is based on the inclusion of a random exchange process in product operator calculations on a microscopic level. This product operator approach can be applied to estimate exchange contributions when using sophisticated pulse sequences that cannot be easily described analytically. The method applies to the full range of exchange times measurable by NMR and can incorporate interference effects between exchange and other processes such as scalar coupling. To demonstrate its utility, simulated relaxation data were compared with theoretical predictions of spin-locking and Carr-Purcell spin-echo sequences with hard and adiabatic pulses, using different time scales for a two site chemical-exchange process. Finally, simulations were used to examine a system in which a second random process is superimposed on a simple two-site exchange process. The method was found to provide a simple and robust tool to analyze pulse sequences and equations commonly used to study exchange-induced relaxation. PMID- 15546761 TI - The set of triple-resonance sequences with a multiple quantum coherence evolution period. AB - The new pulse sequence building block that relies on evolution of heteronuclear multiple quantum coherences is proposed. The particular chemical shifts are obtained in multiple quadrature, using linear combinations of frequencies taken from spectra measured at different quantum levels. The pulse sequences designed in this way consist of small number of RF-pulses, are as short as possible, and could be applied for determination of coupling constants. The examples presented involve 2D correlations HNCO, HNCA, HN(CO)CA, and H(N)COCA via heteronuclear zero and double coherences, as well as 2D HNCOCA technique with simultaneous evolution of triple and three distinct single quantum coherences. Applications of the new sequences are presented for 13C,15N-labeled ubiquitin. PMID- 15546762 TI - Analysis of longitudinal multispin orders in strongly coupled spin systems. AB - Longitudinal multispin orders provide an effective way for measurement of scalar couplings and also to probe molecular interactions and dynamics. Analysis of longitudinal orders has been made in strongly coupled AB and ABX spin systems to determine the dependence of strong coupling parameter on these orders. Experimental and simulated spectra of various longitudinal orders are illustrated for these spin systems. This general procedure can be extended to broad range of spin systems to understand the influence of strong coupling on longitudinal orders. PMID- 15546763 TI - An open volume, high isolation, radio frequency surface coil system for pulsed magnetic resonance. AB - We present an open volume, high isolation, RF system suitable for pulsed NMR and EPR spectrometers with reduced dead time. It comprises a set of three RF surface coils disposed with mutually parallel RF fields and a double-channel receiver (RX). Theoretical and experimental results obtained with a prototype operating at about 100 MHz are reported. Each surface RF coil (diameter 5.5 cm) was tuned to f0=100.00+/-0.01 MHz when isolated. Because of the mutual coupling and the geometry of the RF coils, only two resonances at f1=97.94 MHz and f2=101.85 MHz were observed. We show they are associated with two different RF field spatial distributions. In continuous mode (CW) operation the isolation between the TX coil and one of the RX coils (single-channel) was about -10 dB. By setting the double-channel RF assembly in subtraction mode the isolation values at f1 or f2 could be optimised to about -75 dB. Following a TX RF pulse (5 micros duration) an exponential decay with time constant of about 600 ns was observed. The isolation with single-channel RX coil was about -11 dB and it increased to about 47 dB with the double-channel RX in subtraction mode. Similar results were obtained with the RF pulse frequency selected to f2 and also with shorter (500 ns) RF pulses. The above geometrical parameters and operating frequency of the RF assembly were selected as a model for potential applications in solid state NMR and in free radical EPR spectroscopy and imaging. PMID- 15546764 TI - Use of spatial encoding in NMR photography. AB - NMR photography has gained significant attention as a method of storing and retrieving information using NMR spectroscopy. Among the commonly practiced methods the most important is the frequency encoding by use of a multi-frequency pulse on a liquid crystal molecule. We propose and demonstrate another robust method which relies on spatial encoding. Spatial information is mapped onto the spectrum, if excited and recorded in the presence of a gradient. The encoding is performed by applying a multi-frequency pulse in the presence of a gradient. The subsequent acquisition, under a gradient, helps storing this spatial information on a one-dimensional spectrum. Series of such spectra can also store two dimensional patterns. This procedure is described and exemplified in this paper. PMID- 15546765 TI - Determination of moisture fraction in wood by mobile NMR device. AB - A mobile NMR probe has been used as a non-destructive and non-invasive tool for water content analysis on wood samples. The porosity index, express as the fraction of the sensitivity volume of the probe occupied by water, is here proposed as an alternative to the moisture content index, namely the amount of water mass with respect to the mass of dried sample. In principle the method can be applied to any kind of porous media that has not detectable proton signal from the rigid matrix as, for instance, in building materials. In wood, where proton signal can be detected also from cellulose and others macromolecular components, some considerations and artifices are here proposed for eliminating this contribution. The method has allowed performing moisture volume fraction analysis on wood samples characterized by different wood species, cutting and moisture contents. The NMR data of moisture detection as volume fraction have successfully been compared with those obtained by the gravimetric method. PMID- 15546766 TI - Diagnosis of feline haemoplasma infection in Australian cats using a real-time PCR assay. AB - A total of 147 cats from the Sydney area of Australia that had blood samples submitted to veterinary laboratories were tested using a real-time polymerase chain reaction (PCR) assay able to detect and distinguish the two feline haemoplasma species. This sample number included two cats diagnosed with feline haemoplasma infection by routine blood smear examination. Statistical analysis was performed to evaluate associations between haemoplasma infection, age, sex, breed, haematocrit (HCT) values and anaemia status. One hundred and six cats (72.1%) were negative. Thirty-four cats (23.1%) were positive for 'Candidatus M. haemominutum', six cats (4.1%) were positive for M. haemofelis and one cat (0.7%) was positive for both species. Older, male, non-pedigree cats, with lower HCT values were more likely to be infected with 'Candidatus M. haemominutum'. Significant inverse correlation was found between the amount of M. haemofelis DNA present in the blood and the HCT value. This report documents the existence of, and prevalence of, both haemoplasma species in a sample of cats in Australia and is the first to use quantitative real-time PCR in a prevalence study for haemoplasma infection. PMID- 15546767 TI - Congenital diseases of feline muscle and neuromuscular junction. AB - Although muscle diseases occur relatively rarely in cats, a number of congenital feline myopathies have been described over the last 20 years and are reviewed in this paper. Some of them have been reported exclusively in specific breeds, including the hypokalaemic myopathy of Burmese cats, type IV glycogen storage disease in Norwegian Forest cats, or the myopathy of Devon Rex. Other congenital disorders of muscle and neuromuscular junction such as myotonia congenita, dystrophin-deficient hypertrophic feline muscular dystrophy, laminin alpha2 deficiency, or congenital myasthenia gravis may occur in any cat. A systematic approach is essential in order to efficiently obtain a timely diagnosis in cats showing signs of muscle disease. After a thorough clinical examination, this approach includes blood analyses (eg, serum concentration of muscle enzymes), electrophysiology where available (electromyography, nerve conduction studies), and sampling of muscle biopsies for histological, histochemical and immunohistochemical evaluation. When available, detection of healthy carriers of these genetic disorders is important to eliminate the gene mutations from breeding families. Clinicians regularly receiving feline patients must have a good knowledge of congenital feline myopathies and the features which enable a diagnosis to be made and prognosis given. Besides preserving or restoring the well-being of the myopathic patient, rapid and efficient information and counselling of the breeders are of central importance in order to prevent the recurrence of the problem in specific breeding lines. PMID- 15546768 TI - Congenital phimosis in a cat. AB - An 8-week-old domestic shorthair cat was presented with a history of pollakiuria and polydipsia. No abnormalities were detected during the clinical examination other than the penis could not be extruded from the prepuce. Urine culture demonstrated a growth of E. coli, which was treated with a prolonged course of amoxycillin. The polydipsia resolved and the pollakiuria improved but did not completely abate after antibiotic treatment and the cat had occasional bouts of haematuria. Because of the history of pollakiuria and polydipsia and the ongoing, occasional bouts of haematuria, the cat underwent surgical correction of the congenital phimosis. A small wedge of the dorsal prepuce was removed and the ipsilateral edges of the prepuce were apposed using a simple interrupted pattern. The procedure was quick, easy to perform and led to a resolution of the occasional bouts of haematuria and pollakiuria. PMID- 15546769 TI - Asymptomatic double ureteral stricture in an 8-month-old Maine Coon cat: an imaging-based case report. AB - Congenital ureteral strictures are rare both in human and veterinary medicine. While the diagnosis is generally made after a symptomatic or febrile urinary tract infection, we report here a case in which this condition represented an incidental finding during routine ultrasonography. To the best of the authors' knowledge imaging findings of a double monolateral ureteral stricture have not previously been reported. PMID- 15546770 TI - Necrosis of hippocampus and piriform lobe: clinical and neuropathological findings in two Italian cats. AB - The present paper reports the clinical and neuropathological findings in two cats with a neuropathologically confirmed diagnosis of necrosis of the hippocampus and piriform lobe. The cats were presented because of acute onset of behavioural changes and complex partial seizures. The neurological examination suggested a forebrain lesion. The results of blood examination were within the normal range, and the cerebrospinal fluid (CSF) analysis and computed tomography (CT) scan in one cat did not show any abnormality. Despite therapy with diazepam (Valium; Roche) there was deterioration of the clinical signs and the cats were euthanased. The neuropathological examination revealed hippocampal necrosis that included the piriform lobe. PMID- 15546771 TI - Infections and some other conditions affecting the skin and subcutis of the naso ocular region of cats--clinical experience 1987-2003. AB - Infections of the skin or subcutis of the naso-ocular region develop through two mechanisms. Cases with lesions but without concomitant signs of nasal disease probably result from cat scratch injuries. Under certain circumstances, such lacerations result in the introduction of saprophytic microorganisms in such large numbers that host defence mechanisms are overwhelmed. This results in localised, variably invasive, disease in an otherwise immunocompetent host. An unpredictable range of organisms can give rise to such infections including a variety of fungal and bacterial genera. Causal organisms will likely vary from one geography to another as a result of differences in soil type and related environmental factors. Accordingly, procurement of appropriate tissue specimens for culture and susceptibility testing is essential to guide therapy, as these cases require medical and sometimes surgical intervention in order to effect a favourable outcome. In contrast, patients with naso-ocular lesions and concurrent signs of nasal disease have a different pathogenesis. Primary infection of the sinonasal region likely results from the inhalation of infectious propagules, with the infection subsequently penetrating overlying bones to invade the subcutaneous space. These lesions are typically the result of cryptococcosis or aspergillosis and must be distinguished from invasive nasal malignancies. An approach to the investigation and treatment of these patients is presented together with photographs of representative cases. PMID- 15546772 TI - Paraneoplastic leukocytosis with mature neutrophilia in a cat with pulmonary squamous cell carcinoma. AB - A 13-year old cat had 3-day duration of vomiting, lethargy, and anorexia. A complete blood count revealed a severe neutrophilia (126.9 x 10(3) cells/microl). Thoracic radiographs demonstrated a large solitary lung mass. A bone marrow aspirate documented myeloid hyperplasia. A left intercostal thoracotomy was performed and left cranial and caudal lung lobectomies were performed in order to remove the mass in its entirety. Histological diagnosis was squamous cell carcinoma of the lung. Following surgery, the severe neutrophilia began to decrease. It was in the normal range approximately 6 weeks postoperatively. The presence of a primary lung tumor combined with a mature neutrophilia with resolution following surgical resection suggests a paraneoplastic syndrome. PMID- 15546773 TI - Prostatic carcinoma causing urethral obstruction and obstipation in a cat. AB - A 9-year-old intact male cat was presented for vomiting and straining to defecate. A large abdominal mass was palpated. The urinary bladder was full and non-expressible. Exploratory laparotomy revealed that the mass was compressing the colon and encircling the urethra caudal to the bladder. The mass was removed, the urethra transected, and the urinary bladder marsupialized to the ventral abdominal wall to allow urine drainage. Histopathologic examination of the mass revealed a prostatic carcinoma. The cat died approximately 6 weeks after removal of the mass. This is the first reported case of a prostatic carcinoma causing urethral obstruction and obstipation in a cat. PMID- 15546774 TI - American Association of Feline Practitioners basic guidelines of judicious therapeutic use of antimicrobials in cats (approved by the AVMA Executive Board, June 2001). PMID- 15546775 TI - Magnetoneurography: theory and application to peripheral nerve disorders. AB - Magnetoneurography (MNG) is a non-invasive method to trace and visualize three dimensionally the propagation path of compound action currents (CAC) along peripheral nerves. The basic physical and physiological principle is the mapping of extremely weak magnetic fields generated by the intraaxonal longitudinal ion flows of evoked nerval CAC using SQUID sensors (Superconducting Quantum Interference Devices). During recent years, MNG protocols have been established which allow for a non-invasive spatiotemporal tracing of impulse propagation along peripheral nerves in humans and in particular along proximal nerve segments in a clinical setting. Thereby, the three-dimensional path, the local nerve conduction velocity, the length and strength of the CAC de- and repolarization phase have been reconstructed. First recordings in patients demonstrated that the method is sensitive enough to detect and to localize nerve conduction anomalities along nerve roots, as, e.g. caused by lumbosacral disc herniation. This review on MNG will focus on those studies which provide data from humans and thereby reveal perspectives for its future clinical applications. PMID- 15546776 TI - Recovery of movement-related potentials in the temporal course after prefrontal traumatic brain injury: a follow-up study. AB - OBJECTIVE: The movement-related potential (MRP) is an EEG measure related to self initiated movements, consisting of the Bereitschaftspotential (BP), the negative slope, and the motor potential. Since in a former study the BP was reduced in acute prefrontal traumatic brain injury (TBI) patients, the present study examined the MRPs' course in follow-up examinations. METHODS: Right index finger MRPs of 22 patients with contusions of the prefrontal cortex were recorded 12, 26, and 52 weeks after TBI and compared to controls. RESULTS: Within the patient group, a significant increase of the BP in the temporal course after TBI was observed. MRPs 12 and 26 weeks after TBI did not differ significantly from the control group. One year after TBI, significantly enhanced BPs were found. CONCLUSIONS: In the temporal course after prefrontal TBI, a recovery of the initially reduced BP was observed. The enhanced BP areas 1 year after TBI might represent the need for increased cognitive resources during movement preparation, supporting a recovered, but less effective neuronal network. SIGNIFICANCE: The present study represents the first longitudinal follow-up study of MRPs after prefrontal brain lesion. The observed changes reflect the plastic capacity of the brain, reorganizing the neuronal network function. PMID- 15546777 TI - Changes in arousal level by differential exercise intensity. AB - OBJECTIVE: The purpose of the present study was to investigate the influence of exercise intensity on arousal level. METHODS: Twelve subjects (22-33 years) performed a S1-S2 reaction time task consisting of warning stimulus (S1) and imperative stimulus (S2) in a control condition, and again after low, medium, and high intensity pedaling exercises. During this task, contingent negative variation (CNV) and spontaneous electroencephalogram before S1 were measured as indicators for arousal level. RESULTS: CNV amplitude after high intensity pedaling exercise was significantly smaller than after medium pedaling exercise. Compared to the control condition, relative power value of alpha waves increased after the high intensity exercise. CONCLUSIONS: These results suggested that arousal level was reduced after high intensity exercise and reached a state near optimal level after medium intensity exercise. The findings also suggested that changes in CNV amplitude by differences in exercise intensity followed an inverted-U shaped dose response curve. SIGNIFICANCE: The present study supported the view that CNV amplitude and arousal level followed an inverted-U relationship. It is concluded that differences in exercise intensity influenced arousal level. PMID- 15546778 TI - Quantitative continuous EEG for detecting delayed cerebral ischemia in patients with poor-grade subarachnoid hemorrhage. AB - OBJECTIVE: Delayed cerebral ischemia (DCI) due to vasospasm is often undetected by clinical exam in patients with poor-grade subarachnoid hemorrhage (SAH). The purpose of this study was to identify quantitative EEG (qEEG) parameters that are most sensitive and specific for the detection of DCI in stuporous or comatose SAH patients. METHODS: Of 78 consecutive Hunt-Hess grade 4 or 5 SAH patients admitted to our Neuro-ICU over a 2-year period, 48 were eligible for participation and 34 were enrolled. Continuous EEG monitoring was performed from post-operative day 2 to post-SAH day 14. In each patient, 20 artifact-free, 1 min EEG-clips following an alerting stimulus were analyzed: 10 clips were obtained on monitoring day 1 (baseline), and 10 on days 4-6 (follow-up). In DCI patients, follow-up clips were obtained after the onset of deterioration and before infarction had occurred. Twelve qEEG parameters were calculated using fast Fourier transformation; generalized estimating equations were used to compare ratios of change in qEEG parameters in patients with and without DCI. RESULTS: Nine of 34 patients (26%) developed DCI. The alpha/delta ratio (alpha power/delta power; ADR) demonstrated the strongest association with DCI. The median decrease of ADR for patients with DCI was 24%, compared to an increase of 3% for patients without DCI (Z=4.0, P<0.0001). Clinically useful cut-offs included 6 consecutive recordings with a >10% decrease in ADR from baseline (sensitivity 100%, specificity 76%) and any single measurement with a >50% decrease (sensitivity 89%, specificity 84%). CONCLUSIONS: A decrease in the ADR may be a sensitive method of detecting DCI, with reasonable specificity. This post-stimulation qEEG parameter may supplement the clinical exam in poor-grade SAH patients and may prove useful for the detection of DCI. SIGNIFICANCE: Following ADRs may allow earlier detection of DCI and initiation of interventions at a reversible stage, thus preventing infarction and neurological morbidity. PMID- 15546779 TI - Vinpocetine prevents 4-aminopyridine-induced changes in the EEG, the auditory brainstem responses and hearing. AB - OBJECTIVE: The purpose of the present study was to investigate if the sodium channel blocker and memory enhancer, vinpocetine, was capable to overcome the epileptic cortical activity, the abnormalities in the later waves of the auditory brainstem responses (ABRs) and the hearing loss induced by 4-AP at a convulsing dose in the guinea pig in vivo. METHODS: EEG and ABR recordings before and at specific times within 2h after the injection of 4-AP (2 mg/kg, i.p.) were taken in animals pre-injected i.p. with vehicle or with vinpocetine (2 mg/kg) 1 h before 4-AP. The amplitude and latency of the ABR waves induced by a monoaural stimulus of high intensity (100 dB nHL) at 4 and 8 kHz pure tone frequencies and the ABR threshold were determined in the animals exposed to the different experimental conditions. RESULTS: Vinpocetine inhibited the EEG changes induced by 4-AP for the ictal and post-ictal periods as well as the alterations in amplitude and latency of P3 and P4 and the increase in the ABR threshold induced by 4-AP. CONCLUSIONS: Vinpocetine prevents the retro-cochlear alterations and the hearing decline that accompany the epileptic cortical activity. SIGNIFICANCE: Vinpocetine could be a promising alternative for the treatment of epilepsy. PMID- 15546780 TI - Quantitative visualization of ictal subdural EEG changes in children with neocortical focal seizures. AB - OBJECTIVE: To quantify the ictal subdural electroencephalogram (EEG) changes using spectral analysis, and to delineate the quantitatively defined ictal onset zones on high-resolution 3D MR images in children with intractable neocortical epilepsy. METHODS: Fourteen children with intractable neocortical epilepsy (age: 1-16 years) who had subsequent resective surgery were retrospectively studied. The subjects underwent a high-resolution MRI and prolonged subdural EEG recording. Spectral analysis was applied to 3 habitual focal seizures. After fast Fourier transformation of the EEG epoch at ictal onset, an amplitude spectral curve (square root of the power spectral curve) was created for each electrode. The EEG magnitude of ictal rhythmic discharges was defined as the area under the amplitude spectral curve within a preset frequency band including the ictal discharge frequency, and calculated for each electrode. The topography mapping of ictal EEG magnitude was subsequently displayed on a surface-rendered MRI. Finally, receiver operating characteristic (ROC) analysis was performed to evaluate the consistency between quantitatively and visually defined ictal onset zones. RESULTS: The electrode showing the maximum of the averaged ictal EEG magnitude was part of the visually defined ictal onset zone in all cases. ROC analyses demonstrated that electrodes showing >30% of the maximum of the averaged ictal EEG magnitude had a specificity of 0.90 and a sensitivity of 0.74 for the concordance with visually defined ictal onset zones. SIGNIFICANCE: Quantitative ictal subdural EEG analysis using spectral analysis may supplement conventional visual inspection in children with neocortical epilepsy by providing an objective definition of the onset zone and its simple visualization on the patient's MRI. PMID- 15546781 TI - Seizure incidence during single- and paired-pulse transcranial magnetic stimulation (TMS) in individuals with epilepsy. AB - OBJECTIVE: We reviewed published data and our own data to determine a quantitative incidence of seizure in subjects with epilepsy undergoing single- and paired-pulse transcranial magnetic stimulation (spTMS and ppTMS) and to explore conditions that may increase this risk. METHODS: A PubMed literature search was performed, and articles from this search were reviewed. Subjects from our institution also were included. RESULTS: The crude risk of a TMS-associated seizure ranges from 0.0 to 2.8% for spTMS and 0.0-3.6% for ppTMS. Medically intractable epilepsy and lowering antiepileptic drugs were associated with increased incidence. There was significant center-to-center variability that could not be explained by differences in patient population or by differences in reported stimulation parameters. In all cases, seizures were similar to each subject's typical seizure and without long-term adverse outcome. In most cases, doubt was expressed in the original reports as to whether the seizures were induced by TMS or merely coincidental. CONCLUSIONS: The incidence of seizure in a subject with epilepsy during spTMS and ppTMS appears to be small and not associated with long-term adverse outcome. The incidence is higher under the specific conditions mentioned above. SIGNIFICANCE: These findings may enable researchers to more accurately inform subjects of seizure risk during TMS. PMID- 15546782 TI - EEG dipole source localisation of interictal spikes acquired during routine clinical video-EEG monitoring. AB - OBJECTIVE: We investigated the feasibility of electroencephalography (EEG) dipole source localisation of interictal epileptiform discharges from data acquired during routine clinical inpatient video-EEG monitoring (VEM) and compared a 19 channel 'routine montage' with a 29-channel 'surgical montage' that includes an additional row of 10 inferior temporal electrodes. METHODS: Twenty consecutive patients who had VEM for the presurgical evaluation of medically refractory partial epilepsy were screened. Thirteen of the patients had focal interictal spikes recorded, and in 11 (85%) these were technically satisfactory for source localisation. Fourteen spike foci were analysed as 3 patients had bilateral independent spikes. EEG data was acquired with 29 electrodes including an inferior temporal row (surgical montage). For comparison, the 10 additional electrodes were excluded from analysis (routine montage). Using NEUROSCAN Source 2.0 software, a computed dipole source localisation of averaged spikes was performed utilising a magnetic resonance imaging-based finite element model. Dipole localisation was compared with that of the Comprehensive Epilepsy Program (CEP) evaluation. RESULTS: Using the surgical montage dipole source localisation was consistent with the CEP spike localisation for 13/14 spikes (93%, P<0.005), compared with only 5/14 spikes (36%) using the routine montage. CONCLUSIONS: Data derived from routine clinical inpatient VEM using a routine montage can yield accurate EEG dipole source localisation, but significantly more accurate localisation is obtained using the surgical montage. PMID- 15546783 TI - Classifying EEG-based motor imagery tasks by means of time-frequency synthesized spatial patterns. AB - OBJECTIVE: To develop a single trial motor imagery (MI) classification strategy for the brain-computer interface (BCI) applications by using time-frequency synthesis approach to accommodate the individual difference, and using the spatial patterns derived from electroencephalogram (EEG) rhythmic components as the feature description. METHODS: The EEGs are decomposed into a series of frequency bands, and the instantaneous power is represented by the envelop of oscillatory activity, which forms the spatial patterns for a given electrode montage at a time-frequency grid. Time-frequency weights determined by training process are used to synthesize the contributions from the time-frequency domains. RESULTS: The present method was tested in nine human subjects performing left or right hand movement imagery tasks. The overall classification accuracies for nine human subjects were about 80% in the 10-fold cross-validation, without rejecting any trials from the dataset. The loci of MI activity were shown in the spatial topography of differential-mode patterns over the sensorimotor area. CONCLUSIONS: The present method does not contain a priori subject-dependent parameters, and is computationally efficient. The testing results are promising considering the fact that no trials are excluded due to noise or artifact. SIGNIFICANCE: The present method promises to provide a useful alternative as a general purpose classification procedure for MI classification. PMID- 15546784 TI - The use of QSD (q-sequence deconvolution) to recover superposed, transient evoked responses. AB - OBJECTIVE: We describe q-sequence deconvolution (QSD), a new data acquisition/analysis method for evoked-responses that solves the problem of waveform distortion at high stimulus repetition-rates, due to response overlap. QSD can increase the sensitivity of clinically useful evoked-responses because it is well known that high stimulus repetition-rates are better for detecting pathophysiology. METHODS: QSD is applicable to a variety of experimental conditions. Because some QSD-parameters must be chosen by the experimenter, the underlying principles and assumptions of the method are described in detail. The theoretical and mathematical bases of the QSD method are also described, including some equivalent computational formulations. RESULTS: QSD was applied to recordings of the human auditory brainstem response (ABR) at stimulus repetition rates that overlapped the responses. The transient ABR was recovered at all rates tested (highest 160/s), and showed systematic changes with stimulus repetition rate within a single subject. CONCLUSIONS: QSD offers a new method of recovering brain evoked-response activity having a duration longer than the time between stimuli. SIGNIFICANCE: The use of this new technique for analysis of evoked responses will permit examination of brain activation patterns across a broad range of stimulus repetition-rates, some never before studied. Such studies will improve the sensitivity of evoked-responses for the detection of brain pathophysiology. New measures of brain activity may be discovered using QSD. The method also permits the recovery of the transient brain waveforms that overlap to form 'steady-state' waveforms. An additional benefit of the QSD method is that repetition-rate can be isolated as a variable, independent of other stimulus characteristics, even if the response is a nonlinear function of rate. PMID- 15546785 TI - Repetitive nerve stimulation in myasthenia gravis--relative sensitivity of different muscles. AB - OBJECTIVE: To correlate repetitive nerve stimulation (RNS) decrement in different muscles with the predominant clinical presentation in myasthenia gravis (MG), and to study single fibre EMG (SFEMG) sensitivity in ocular MG. METHODS: Sixty-nine, untreated, consecutive patients suspected for MG were observed prospectively for a minimum of 6 months. Those who improved on medical treatment were diagnosed as MG. The others, in whom the neurophysiological studies were normal and that did not improve on medical treatment served as a control group, from which normative data for RNS and SFEMG was obtained. The MG patients were further classified in 3 subgroups according to the predominant clinical presentation: group I (ocular); group b (bulbar); and group a (axial). We performed RNS in nasalis, trapezius, anconeus, and abductor digiti minimi. All patients with ocular MG underwent jitter determination of the orbicularis oculi muscle. RESULTS: Thirty-seven patients were diagnosed as MG (group I, 15; group b, 13; group a, 9). In group I, RNS was abnormal in 33% of the patients. RNS studies disclosed at least one abnormal muscle response in every patient in groups a and b. Trapezius was significantly more sensitive in group a, and anconeus and nasalis in group b (P < 0.01). Jitter was abnormal in all patients in group I, and the most sensitive parameter was an increased number of unstable pairs, 100%. CONCLUSIONS: Based on these observations, we recommend that a shoulder muscle, as the trapezius, should be studied first in the limb-axial presentation of MG, and the anconeus-nasalis muscles in predominant bulbar MG. In ocular MG, RNS is not sensitive and jitter should be performed in facial muscles. SIGNIFICANCE: This paper shows the unequal sensitivity of several muscles to RNS in different forms of MG. PMID- 15546786 TI - Does retrograde axonal atrophy really occur in carpal tunnel syndrome patients with normal forearm conduction velocity? AB - OBJECTIVE: The cause of decreased median forearm motor conduction velocity (FMCV) in carpal tunnel syndrome (CTS) is best ascribed to retrograde axonal atrophy (RAA); however, the relationships between the occurrence of RAA and electrophysiological or clinical severity remains controversial. We attempt to determine whether RAA really occurs in CTS patients with normal median FMCV and to investigate any relationships between RAA and severity of compression at the wrist. METHODS: Consecutive CTS patients were enrolled and age-matched volunteers served as controls. We performed conventional nerve conduction studies (NCS) and measured median and ulnar distal motor latencies (DML), FMCV, compound muscle action potential (CMAP) amplitudes, distal sensory latencies (DSL), and sensory nerve action potential (SNAP) amplitudes. Furthermore, palmar median stimulation was done to calculate the wrist-palm motor conduction velocity (W-P MCV). Patients included for analysis should have normal FMCV and needle examination. We compared each electrodiagnostic parameters between the patient group and controls. RESULTS: The mean+/-SD of the W-P MCV for patients and controls were 33.26+/-6.74 and 52.14+/-5.85 m/s and those of median FMCV were 55.26+/-3.56 and 57.82+/-3.9 m/s, respectively. There was a significant reduction in the W-P MCV (36.2%, P<0.00001), significant decrease in the median FMCV (4.43%, P<0.00001) and SNAP amplitudes, and an increase of the DML and DSL in the patient group (P<0.00001) compared to the controls; however, there were no differences in median and ulnar CMAP amplitudes, ulnar FMCV and DML between the controls and patients. CONCLUSIONS: RAA and relatively slowed median FMCV do occur in CTS patients with normal median FMCV, regardless of severity of clinical manifestations and electrophysiological abnormalities. SIGNIFICANCE: This article provides new information for research of the electrophysiological changes of the proximal nerve part at distal injury. PMID- 15546787 TI - Modulation of ankle muscle postural reflexes in stroke: influence of weight bearing load. AB - OBJECTIVE: Given the known sensorimotor deficits and asymmetric weight-bearing posture in stroke, the aim of this study was to determine whether stroke affects the modulation of standing postural reflexes with varying weight-bearing load. METHODS: Ten individuals with chronic stroke and 10 healthy older adult controls were exposed to unexpected forward and backward platform translations while standing. Three different stance conditions were imposed: increased weight bearing load, decreased weight-bearing load, and self-selected stance. Surface electromyography from bilateral ankle dorsiflexors (tibialis anterior) and extensors (gastrocnemius) were recorded and the magnitude of background muscle activity (prior to the platform translation) and postural reflex onset latency and magnitude (75 ms following reflex onset) were determined. RESULTS: Load modulation of ankle extensors was found in controls and individuals with stroke. Although controls demonstrated modulation of ankle dorsiflexors to different loads, individuals with stroke did not show this modulation. Further, load did not change the onset latency of postural reflexes of the individuals with stroke. CONCLUSIONS: The delayed paretic muscle onset latencies in conjunction with impaired modulation of ankle dorsiflexor postural reflexes may contribute to the instability and frequent falls observed among individuals with stroke. SIGNIFICANCE: The results provide some insight into standing postural reflexes following stroke. PMID- 15546788 TI - Expansion of nociceptive withdrawal reflex receptive fields in spinal cord injured humans. AB - OBJECTIVE: In spinal cord injured (SCI) subjects, exaggerated withdrawal reflexes associated with a dominant flexor pattern irrespective of stimulation site have been reported. In the present study, withdrawal reflex receptive field (RRF) was determined in complete SCI subjects (N=9). METHODS: Distributed electrical stimulation was applied to the sole of the foot, and reflexes in tibialis anterior, soleus, biceps femoris, and vastus lateralis muscles were recorded together with knee and ankle movement trajectories. A group of spinally intact subjects (N=10) were included as controls. With the subjects in supine position, stimulation was applied to 10 different sites on the foot sole. Based on the tibialis anterior reflex threshold for stimulation on the mid foot sole, two stimulus intensities (1.1 times the reflex threshold and 1.4 times the reflex threshold) were used for all 10 sites. RESULTS: In SCI subjects, dorsi-flexion dominated independent of stimulus site and the tibialis anterior RRF covered the entire foot sole in contrast to a well-defined tibialis anterior receptive field at the medial, distal foot sole in the spinally intact subjects. Further, the soleus RRF also covered the entire sole in the SCI subjects. The reflexes in biceps femoris and vastus lateralis muscles were small and associated with weak knee flexion at all 10 sites in the SCI subjects and in the controls. CONCLUSIONS: The RRF of the ankle flexor and the ankle extensor muscles both covered the entire sole of the foot indicating an expansion of the RRFs following spinal cord injury. The expansion is most likely due to lack of descending inhibitory control and/or increased sensitivity of the spinal reflex loop in the SCI subjects. SIGNIFICANCE: The study improves the understanding of spinal reflex control in spinal intact and spinal cord injured subjects. PMID- 15546789 TI - Modeling the relationship between psychophysical perception and electrically evoked compound action potential threshold in young cochlear implant recipients: clinical implications for implant fitting. AB - OBJECTIVE: In cochlear implant recipients, the threshold of the electrically evoked compound action potential (ECAP) has been shown to correlate with the perceptual detection threshold and maximum comfortable loudness levels (respectively, T- and C-levels) used for implant programming. Our general objective was to model the relationship between ECAP threshold and T/C-levels by taking into account their relative changes within each subject. In particular, we were interested in investigating further the validity of ECAP threshold as a predictor of psychophysical levels, depending on intra-cochlear electrode location and time of testing (from 1 to 18 months post-implantation). METHODS: A total of 370 ECAP thresholds, measured in 49 children, using a Nucleus 24 cochlear implant, were compared with the corresponding T- and C-levels obtained at the same visit, for the same electrode. Response profiles for the whole group of patients were modeled across four test electrodes spaced equally along the electrode array from base towards apex. A linear regression model was constructed and the quality of the ECAP threshold-based predictions was assessed by testing for correlation between measured and predicted psychophysics. Comparison was made with a more simplistic model (described here as the 'parallel profiles method') stipulating, within each subject, a 1 microA increase in psychophysical levels for every 1 microA increase in ECAP threshold. RESULTS: Offset between ECAP threshold and psychophysics profiles was found to vary significantly along the electrode array for the T-, but not for the C-level. In contrast with the parallel profiles method, our regression model predicted, within each subject, an average increase of 0.23 microA (95% confidence interval: 0.18-0.28) in T-level for every 1 microA increase in ECAP threshold. This correction improved the quality of T-level prediction when our model was run using measured T-level and ECAP threshold from a reference electrode (r=0.77 vs. r=0.62). The shorter the distance between the electrode for which T-level was predicted and the one used as reference, the stronger the correlation between measured and predicted T levels. In addition, poorer T-level predictions were obtained at the basal end of the array during the first 3 months post-implantation. In contrast to T-level, individual changes in C-level with ECAP threshold exhibited heterogeneous patterns across subjects so that no common coefficient could account for these changes. However, applying the parallel profiles method led to high-quality C level prediction. CONCLUSIONS AND SIGNIFICANCE: The results suggest that covariation between ECAP thresholds and psychophysics plays a decisive role in the relationship of ECAP threshold with T-, but not with C-level. Therefore, our regression model and the parallel profiles method should both be used for predicting, respectively, the T- and the C-levels. Although the predictability of our regression model seems to be better for middle and apical electrodes, its utilization should be extended to basal electrodes after 6 months' implant use. PMID- 15546790 TI - Effects of total sleep-deprivation on waking human EEG: functional cluster analysis. AB - OBJECTIVE: The purpose of the present study was to investigate the effects of total sleep deprivation (TSD) on brain functions with an identification procedure for strongly interactive brain regions, relying on functional cluster (FC) analysis in multichannel electroencephalogram (EEG) data. METHODS: EEGs from 16 electrodes in 18 healthy, right-handed, young male volunteers were recorded before TSD (after normal sleep) and after 24 h of experimentally induced sleep deprivation. We estimated cluster index to characterize joint interactions among many brain regions in order to determine if a particular FC is present or not, and if so, its anatomy. RESULTS: As a result, we confirmed the presence of FC and found different FC patterns in both before and after TSD. The C3 and F7 locations were outside the cluster under the TSD condition, but belonged to the cluster with C4 and F8 before the TSD condition, and the F3/F4, and O1 locations were new entries to the functional cluster during sustained wakefulness. CONCLUSIONS: These results indicate that the neuronal activities of the C3 and F7 location are functionally unrelated, whereas the F3/F4 locations are functionally involved with the C4, F8, and O1 locations after 24 h TSD. Our results suggest that FC changes with elapsed awake time and reflects the change of brain function due to TSD. SIGNIFICANCE: This paper shows the existence of FC both before and after TSD, and the anatomy of each FC is different. So FC analysis would be a potential tool to investigate the simultaneous neuronal activity of human EEGs. PMID- 15546791 TI - Some remarks on the use of motion VEPs to assess magnocellular sensitivity. PMID- 15546793 TI - T cell tolerance and autoimmunity. AB - CD4 T cells are the master controllers of immune responses to protein antigens, and many autoimmune diseases are thought to arise from a breakdown of immunological tolerance in CD4 cells. Peripheral tolerance in CD4 T cells is maintained by several mechanisms, including functional anergy, deletion (death) by apoptosis and suppression by regulatory T lymphocytes (Treg). Using transgenic mouse models, we have explored the roles of these mechanisms in tolerance to cell associated tissue-restricted self-antigens and secreted systemic self-antigens. Tolerance to a membrane form of the antigen expressed in islet beta cells is maintained by Treg, which block T cell differentiation into pathogenic effectors, and by CTLA-4, which increases the activation threshold of T cells and prevents responses to the self-antigen. A systemically produced soluble form of the antigen induces rapid T cell anergy followed by deletion. The induction of anergy does not require either CTLA-4 or Treg, although in the absence of Treg tolerance can be broken more readily by potent immunogenic signals. Encounter with circulating antigen in T cells induces a state of antigen receptor "desensitization" that is associated with a block in proximal receptor-triggered signals. Thus, different mechanisms play dominant roles in T cell tolerance to different types of self-antigens. PMID- 15546794 TI - Autoimmune myocarditis: cellular mediators of cardiac dysfunction. AB - Immune mediators play a critical role in the pathogenesis and outcomes of a number of cardiac diseases. This review summarizes recent findings on the composition of the inflammatory infiltrate and the role of different types and subtypes of immune cells and their products in mediating cardiac dysfunction in experimental autoimmune myocarditis (EAM). CD4+ T cells are required for initiation of myocarditis and their numbers in the heart infiltrate correlate with systolic dysfunction during disease progression. Other immune cells, including CD8+ T cells, granulocytes, and mast cells, can directly affect cardiomyocyte function. When regulatory mechanisms fail, the local damage leads to cardiomyocyte death, replacement fibrosis and overall cardiac dysfunction. EAM provides insights into the role of the immune system in the development of dilated cardiomyopathy (DCM) and heart failure and may serve as a general paradigm for autoimmune organ-specific tissue damage. PMID- 15546795 TI - Autoimmune epitopes: autoepitopes. AB - The identity of reactants for autoantibodies has been successively refined from whole cellular organelles (immunofluorescence), identified molecules (immunoblot; gene expression libraries), epitope regions (truncated cDNAs; peptide scanning) to contact residues, as described here. Most autoantibodies react with conformational epitopes, in which amino acids distant in the linear sequence come into contiguity by protein folding. Identification of contact sites with the antibody paratope requires particular technologies, crystallography, or antibody screening of phage-displayed random peptide libraries. The latter is illustrated by our studies on the autoepitope for anti-PDC-E2 (AMA) in primary biliary cirrhosis (PBC), anti-GAD65 in type 1 diabetes, and anti-C1 of type II collagen in collagen-induced arthritis. More precise definition of the structure of conformational autoepitopes could (a) clarify controversial aspects of autoimmunity including epitope mimicry, epitope spreading, and molecular spatial relationships between B and T cell autoepitopes, and (b) impact on novel diagnostic and therapeutic (vaccine) molecules. PMID- 15546796 TI - Primary biliary cirrhosis: does X mark the spot? AB - Primary biliary cirrhosis (PBC) is an autoimmune disease of unknown etiology leading to progressive destruction of intrahepatic bile duct, with cholestasis, cirrhosis, and eventually liver failure. Epidemiological data indicate that environmental factors trigger autoimmunity in genetically susceptible individuals, although no definitive association of PBC with specific genes has been found. Further, no convincing explanation has been provided for the strong female predominance observed in the prevalence of PBC. However, we recently suggested that the enhanced monosomy X in peripheral white blood cells, and particularly in lymphocytes, of affected women might play a role in the induction of PBC. Such observations appear independent from the degree of cholestasis and specific for PBC. In this review we discuss the implications of these findings and their immunological implications. PMID- 15546797 TI - The immune response to cell death in SLE. AB - Systemic lupus erythematosus (SLE) is a prototypic autoimmune disease characterized by the production of antinuclear antibodies (ANA). These antibodies target a wide variety of antigens whose presence in an immunologically active form may result from cell death processes that cause their translocation and release from cells. As indicated by in vivo model systems, the release of DNA from cells may not be a simple consequence of cell death but rather may require the intervention of other cell types including macrophages. Thus, in mice, administration of either apoptotic or necrotic cells produces a blood DNA response, whereas mice lacking macrophages fail to show blood DNA under the same conditions. Furthermore, the circulating DNA arising from apoptotic and necrotic cells displays a similar pattern with respect to size distribution, with both showing DNA laddering, a pattern indicating enzymatic cleavage. Since circulating DNA in the form of immune complexes can play a role in lupus pathogenesis, these findings suggest that the generation and clearance of dead cells are important events that may underlie autoimmunity in this disease and may be targeted for therapy. PMID- 15546798 TI - Gene expression profiling in the study of the pathogenesis of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with a complex pathogenesis involving multiple genetic and environmental contributions. DNA microarray technology has recently been applied to unravel some of this complexity through genomewide profiling. Early studies using microarray analysis of peripheral blood mononuclear cells (PBMCs) from SLE patients revealed dysregulation of inflammatory cytokines, chemokines, and immune response-related genes, as well as genes involved in apoptosis, signal transduction, and the cell cycle. More recently, using arrays containing many more genes, 4 independent research groups have found that interferon (IFN)-regulated genes are highly overexpressed in the peripheral blood and kidney glomeruli of SLE patients, supporting a crucial role for interferon in SLE. Future studies focusing on target tissues or organs in lupus, including the kidney, may further contribute to our understanding of lupus pathogenesis while providing new targets for therapy. PMID- 15546799 TI - Innate immunity in the antiphospholipid syndrome: role of toll-like receptors in endothelial cell activation by antiphospholipid antibodies. AB - Antiphospholipid antibodies are mainly directed against beta 2 glycoprotein I (beta2GPI), a plasma phospholipid-binding protein expressed on endothelial cells of different anatomical localizations. Anti-beta2GPI antibodies recognize the molecule on endothelial monolayers in vitro, and, once bound, might activate the cells both in vitro and in vivo experimental models inducing a proinflammatory and a procoagulant phenotype. Cell activation is associated with nuclear factor kappaB (NF-kappaB) translocation and with a signaling cascade comparable to that triggered by the toll-like receptors (TLRs)-4. The cell membrane receptor(s) for beta2GPI adhesion is still under investigation. It has been suggested that beta2GPI might adhere through electrostatic interaction between its cationic phospholipid binding site and anionic structures on the cell membrane; however, binding to annexin II-the endothelial cell receptor for tissue plasminogen activator-plays also a role. Because annexin II does not display any transmembrane protein, it has been suggested that it requires a yet unknown "adaptor" protein to signal the cells. Because of the molecular mimicry between beta2GPI and viral/bacterial structures-the natural ligands for TLRs-antibodies might cross-link the molecule associated to annexin II and TLR-4 eventually triggering the signaling. PMID- 15546800 TI - Dysfunctional B cells in systemic lupus erythematosus. AB - The classical view of B cells in the biology of autoimmune responses to infectious and self-antigens (Ag) that they promote immunity primarily by producing antibodies (Ab) is far from being complete. Indeed, studies over the last decade suggest that B cells have extraordinarily diverse functions within the immune system other than Ab production, which could contribute to autoimmunity. They normally play a role in the development of lymphoid architecture, regulating dentritic cells (DC) and T cell subsets function through cytokine production, and in activation of T cells. Receptor editing is also important in B cells which aids in immunity to infection and, possibly, prevention of autoimmunity. Both abnormalities in the distribution of B cells subsets and clinical benefit response to B cell depletion in autoimmune diseases, including systemic lupus erythematosus (SLE), highlight their pivotal function. Transgenic (Tg) animal models have shown that sensitivity of B cells to B cell Ag receptor (BCR) cross-linking is correlated to autoimmunity. Indeed, negative signaling by CD5 and other molecules, such as CD22, in maintaining tolerance through recruitment of src-homology two domain-containing protein tyrosine phosphatase-1 (SHP-1) has also been documented. In fact, we have now reached a newer area whereby B cells returned as an important contributor to autoimmune disorders. PMID- 15546801 TI - Limited clonality in autoimmunity: drivers and regulators. AB - The available T cell repertoire directed against self is appreciable owing to the escape of many clones from negative selection, largely because many determinants on self proteins are cryptic and not presented adequately. In addition, the degeneracy of T cell receptor specificity permits each lymphocyte a broad recognitive potential. Within the available self-reactive repertoire are T cells with high affinity, and these can compete favorably with other T cells with the same specificity. We have studied a "driver clone" and its two specific regulators in the B10.PL model of experimental autoimmune encephalomyelitis and found that each of these repertoires is highly limited. There is a single major clonal family comprising the aggressive driver population, which is public and of high affinity, and just one other minor public clonotype. The receptors of this Vbeta8.2/Jbeta2.7 driver are presented to a CD4 regulator and a CD8 suppressor, each of limited clonality, the latter killing the driver clone by apoptosis, completing a feedback control loop. This tightly regulated group of three cell types furnishes an excellent example of the immune homunculus. PMID- 15546802 TI - Autoimmunity, anergy, lentiviral immunity and disease. AB - Autoimmune antibodies and autoimmune responses have been characterized in both human HIV infection and the rhesus macaque (RM) non-human primate model of SIV infection and reasoned to contribute to the pathogenesis of AIDS. Many theories for the induction and maintenance of such responses have been entertained including molecular mimicry between HIV proteins and self molecules, CD4+ T cell loss accompanied by loss of normal immune regulation that dictate self-non-self reactivity, defective negative/positive selection of T cells to name a few. The precise mechanisms that lead to such immune dysfunction is difficult to study in humans. Our lab has been studying such autoimmune responses in both SIV-infected RM and sooty mangabeys (SM), a species from Africa that are naturally infected with SIV but do not display any detectable signs of immune deficiency or autoimmunity. We submit that this model is an important model since it allows for narrowing down those mechanisms and pathways that are a result of lentiviral infection per se from those that specifically cause disease including autoimmunity. During the course of these studies, we have ruled out a role for plasma and cellular viral loads, anti-viral humoral responses and a variety of cell signaling pathways. We have identified select pathways that appear to play roles in the pathogenesis of lentiviral infection and disease. These include pathways involved in innocent bystander killing by apoptosis of CD4+ T cells, role for differential regulation of the cell cycle, and a role for distinct host proteins that get incorporated by the virions as they are assembled and either bud out of CD4+ T cells or exit the cells in the form of multi-vesicular endosomal particles from monocytes/macrophages from SIV-infected disease susceptible RM and disease-resistant SM. We present our current working model and hypotheses that are designed to elucidate differences that are responsible for such distinct outcomes of lentiviral infection, autoimmunity and disease. We believe that such findings have important implications for the design of vaccines against human HIV infection. PMID- 15546803 TI - Strategies using functional genomics in rheumatic diseases. AB - For functional genomics of inflammatory disorders and infection, rheumatic diseases offer unique features to analyse the transition from infection to chronic inflammation, autoimmunity and immunopathology, both systemic and tissue specific. The diseases are frequent and of considerable socio-economic impact. Well-defined cohorts of patients are available. The tissues and cells involved are readily accessible for molecular analysis. Both genetic predisposition and infection are involved in the aetiopathogenesis of rheumatic diseases. The number of susceptibility and severity genes has been estimated to be at least 30, but only few of them have been identified so far. There is an urgent need for developing new therapies adapted to genetic risk and based on a functional genetic and molecular understanding of chronic inflammation. It is evident that gene analysis in inflammatory rheumatic diseases will not only be beneficial for the large number of patients involved, but will also lead to a better understanding of other inflammatory disorders, thereby possibly leading to novel diagnostic and therapeutic strategies in this important group of disorders. PMID- 15546804 TI - Drug discovery and chemokine receptor antagonists: eppur si muove! AB - The blockade of leukocyte migration has been demonstrated to be a valid option for the treatment of several autoimmune diseases. Chemokines play an active role in regulating cell infiltration into inflammatory sites and disrupting chemokine receptor interactions has emerged as an alternative therapeutic approach. Pharmaceutical companies have developed an intense activity in the drug discovery of chemokine receptor antagonists in the last 10 years. Potent and selective compounds have been obtained and some of them are currently being evaluated in the clinic. The success of these trials will demonstrate whether the blockade of a single receptor is of therapeutic benefit. Alternative approaches, such as pan receptor antagonists or inhibitors of the signalling pathways evoked by chemokines, are also being explored. In the meantime, new relationships between chemokines and receptors will be revealed, increasing our knowledge of such a fascinating field. PMID- 15546805 TI - Is autism an autoimmune disease? AB - Autism spectrum disorder (ASD) is a spectrum of behavioral anomalies characterized by impaired social interaction and communication, often accompanied by repetitive and stereotyped behavior. The condition manifests within the first 3 years of life and persists into adulthood. There are numerous hypotheses regarding the etiology and pathology of ASD, including a suggested role for immune dysfunction. However, to date, the evidence for involvement of the immune system in autism has been inconclusive. While immune system abnormalities have been reported in children with autistic disorder, there is little consensus regarding the nature of these differences which include both enhanced autoimmunity and reduced immune function. In this review, we discuss current findings with respect to immune function and the spectrum of autoimmune phenomena described in children with ASD. PMID- 15546806 TI - Laparoscopic procedures as a risk factor of deep venous thrombosis, superficial ascending thrombophlebitis and pulmonary embolism--case report and review of the literature. AB - Since its introduction laparoscopic surgery has been used for many indications, e.g., cholecystolithiasis, hernia, appendicitis, fundoplication, benign large bowel disease and gynaecological disorders. It has been considered as safe and efficient procedure for most patients with only few contraindications, mostly heart-lung disease. When the initial enthusiasm has been replaced by a more critical observation, more complications of laparoscopy or laparoscopic surgery were not only discovered but also reported. In laparoscopic hernia repair there is a tendency for severe complications when compared to open surgery. There is a controversy on possible side-effects of laparoscopic surgery, e.g., thrombosis, and the increased necessity of prophylaxis for thromboembolic events. Recently a growing number of reports on thromboembolic complications in association with laparoscopic surgery were published. Thrombosis may be caused by detrimental effects of pneumoperitoneum on venous flow (increased abdominal pressure and negative Trendelenburg position) and activation of the haemostatic system. Further risk factors may contribute to the risk to develop venous thrombosis. It is well accepted that varicose veins are associated with an increased risk for the thrombosis. However, the association of varicose veins with complications of laparoscopic surgery is unclear. The possible impact of thrombotic complications makes an analysis of the association of varicose veins or a history of deep vein thrombosis on the development of thrombosis after laparoscopic surgery mandatory. Although this is the first report on ascending thrombophlebitis and thrombosis of the sapheno-femoral junction after laparoscopic surgery, the incidence of deep vein thrombosis or superficial thrombophlebitis after laparoscopic surgery or laparoscopy may be much higher according to the pathophysiological changes during and after these procedures. In many patients venous thrombosis may not be recognized or it appears when the patient is already discharged. CONCLUSION: Laparoscopy and laparoscopic procedures may have an increased risk for the development of thrombosis due to increased abdominal pressure and negative Trendelenburg position. Patients with varicose veins and a history of thromboembolism may aggravate laparoscopy associated risks for the development of thromboembolic complications. Superficial thrombophlebitis in the thigh is not a benign disease entity and may lead to deep vein thrombosis (DVT) and pulmonary embolism (PE). Urgent surgical treatment (high ligation) may be warranted together with low-molecular weight heparin (LMWH) and compressions therapy. Patients with varicose veins and a history of venous thrombosis may not be suitable candidates for laparoscopic surgery. Family practitioners may be confronted with this complication more often since patients are discharged earlier from hospital after laparoscopic interventions due to legislative regulations. PMID- 15546807 TI - Effects of lozenge containing lavender oil, extracts from hops, lemon balm and oat on electrical brain activity of volunteers. AB - Within a randomized double blind, placebo controlled trial the electrical activity of the human brain has been monitored using charge mode technology (Laplacian estimates) after exposure to a lozenge containing 4 different herbal preparations (lavender oil, extracts from hops, lemon balm and oat) or a matching placebo without any active ingredients. Sixteen healthy volunteers (8 males and 8 females) were tested within a crossover design. After baseline recording each subject sucked a lozenge and 2 hours later a second one. Recording was performed immediately after finishing the lozenge and in hourly intervals thereafter. Comparison to reference periods of 10 min eyes open and 5 min eyes closed, respectively, revealed increases in alpha 1, alpha 2 and beta 1 electrical power at the electrode positions Cz, P3, T3 and T5 which were even more pronounced after a second application two hours later. Since alpha 1 changes repeatedly have been attributed to attentional states, increases of this electrical activity must be seen as indicator of a relaxational psychophysiological state. Changes in the alpha2 frequencies have been related to working memory indicating that an increase can be seen as a correlate for attenuating this circuit. Increases of beta1 activity have been seen in the presence of anxiolytic drugs including major and minor tranquilizers. The changes as observed after the application of this herbal composition are therefore in line with the idea of having induced a state of relaxation and regeneration. This interpretation suggests that one could expect from the ingestion of this lozenge to better cope with psychological and emotional stress. The data are further proof that recording computer aided quantitative EEG is a very fruitful and promising approach in psychophysiology. PMID- 15546808 TI - In vitro study on dental erosion provoked by various beverages using electron probe microanalysis. AB - Tooth erosion is often based on chemical processes, among others the use of soft drinks or diverse beverages. The aim of this in vitro study was to analyse the erosive potential of different acidic beverages. Over a time span of 6 hours, dental slices (n=6 slices per tooth) from fully retained wisdom teeth were incubated with different beverages (coca cola, ice tea with lemon, apple juice and white wine). The controls were incubated with a 0.9% sodium chloride solution under the same conditions (37 degrees C, humidified atmosphere of 5% CO2 and 95% air). The quantitative elementary analysis for calcium, phosphorus, oxygen and other trace elements in the dental slices in various depths ranging from 5 to 50 microm was carried out using an electron probe micro-analyser (Jeol JXA 8900RL). A beverage-induced loss of minerals, particularly of the 2 main components calcium and phosphorus, especially in the uppermost layers of the enamel down to a depth of 30 microm could be observed. In the depth of 10 microm, the following total mineral loss could be determined: white wine (16%), coca cola (14.5%), apple juice (6.5%) and ice tea with lemon (6.5%). A direct correlation between the loss of minerals and the pH value of the beverages was not observed, because of the buffering effect of the drinks. The conversion of the weight percentages from the chemical analysis of Ca and P to their atomic percentages showed that during erosion the 2 main components were not dissolved in significantly different percentages. In this study the erosive potential of the tested soft drinks and other beverages could be demonstrated. However, it must be considered that numerous modifying factors influence the enamel surface, so an extrapolation from the in vitro study to an in vivo situation can only be applied with caution. PMID- 15546809 TI - Diagnostic accuracy of atypical p-ANCA in autoimmune hepatitis using ROC- and multivariate regression analysis. AB - INTRODUCTION: Antineutrophil cytoplasmic antibodies (atypical p-ANCA) are detected at high prevalence in sera from patients with autoimmune hepatitis (AIH), but their diagnostic relevance for AIH has not been systematically evaluated so far. METHODS: Here, we studied sera from 357 patients with autoimmune (autoimmune hepatitis n=175, primary sclerosing cholangitis (PSC) n=35, primary biliary cirrhosis n=45), non-autoimmune chronic liver disease (alcoholic liver cirrhosis n=62; chronic hepatitis C virus infection (HCV) n=21) or healthy controls (n=19) for the presence of various non-organ specific autoantibodies. Atypical p-ANCA, antinuclear antibodies (ANA), antibodies against smooth muscles (SMA), antibodies against liver/kidney microsomes (anti-Lkm1) and antimitochondrial antibodies (AMA) were detected by indirect immunofluorescence microscopy, antibodies against the M2 antigen (anti-M2), antibodies against soluble liver antigen (anti-SLA/LP) and anti-Lkm1 by using enzyme linked immunosorbent assays. To define the diagnostic precision of the autoantibodies, results of autoantibody testing were analyzed by receiver operating characteristics (ROC) and forward conditional logistic regression analysis. RESULTS: Atypical p-ANCA were detected at high prevalence in sera from patients with AIH (81%) and PSC (94%). ROC- and logistic regression analysis revealed atypical p-ANCA and SMA, but not ANA as significant diagnostic seromarkers for AIH (atypical p-ANCA: AUC 0.754+/-0.026, odds ratio [OR] 3.4; SMA: 0.652+/-0.028, OR 4.1). Atypical p-ANCA also emerged as the only diagnostically relevant seromarker for PSC (AUC 0.690+/-0.04, OR 3.4). None of the tested antibodies yielded a significant diagnostic accuracy for patients with alcoholic liver cirrhosis, HCV or healthy controls. CONCLUSIONS: Atypical p-ANCA along with SMA represent a seromarker with high diagnostic accuracy for AIH and should be explicitly considered in a revised version of the diagnostic score for AIH. PMID- 15546810 TI - The impact of topical antiseptics on skin microcirculation. AB - AIM: Antiseptics are commonly used in clinical practice to disinfect tissue and to avoid infections. However, topical antiseptics are assumed to have an influence on skin microcirculation, per se. Thus, the aim of the study was to analyse the influence of topically applied antiseptics on the microcirculation of intact skin in vivo. MATERIALS AND METHODS: The investigation was carried out on ears of male hairless mice (SKH-1hr, n = 25). The influence of four antiseptics was examined. Sodium chloride 0.9% served as control. An alcohol-based solution with a mixture of ethanol, 2-propanol and purified water (Softasept), an antiseptic with octenidine dihydrochloride and phenoxyethanol as the main active agents (Octenisept), as well as hexamethylenbiguanide (Lavasept) and 70% ethanol were tested. Intravital fluorescence microscopy in combination with intravenous injection of the fluorescence dyes FITC-Dextran as plasma marker and Rhodamine 6G (leukocyte staining) allowed a quantitative analysis of standard microcirculatory parameters (vessel diameter, functional capillary density, red blood cell velocity, FITC-leakage and leukocyte endothelium interaction). Recordings of the microcirculation in several regions of interest (ROI) were made prior to application and after 10 min exposure time and 60 min after the baseline data. Data were evaluated off-line with aid of computer assisted analysis. RESULTS: The diameter of arterioles decreased after the treatment with the alcoholic solutions. The other two antiseptics (Octenisept and Lavasept) caused a significant increase. Functional capillary density (FCD) was significantly reduced after application of ethanol and Softasept. There was no reduction of FCD following application of Octenisept. After treatment with ethanol and Softasept there was a significant decrease in red blood cell velocity (RBCV). The use of Lavasept revealed a decrease of FCD and RBCV. In the Octenisept treated group RBCV shows a mild increase after 10 minutes. The application of ethanol, Softasept and Lavasept was characterized by a significant increase of leukocyte endothelium interaction (LEI). After treatment with saline and Octenisept LEI remained constant. All used antiseptics except of Octenisept caused a significant leakage of FITC-Dextran. CONCLUSION: The antiseptics used in this study all showed an influence on skin microcirculation. As expected, our findings show that the alcoholic solutions are most aggressive to skin microcirculation. PMID- 15546811 TI - Increased intestinal permeability in patients with inflammatory bowel disease. AB - Intestinal permeability can be measured by the sugar absorption test. This test is based on determining the ratio of the urinary excretion of a large and a small carbohydrate after oral administration. The aim of this study was to determine which combination of carbohydrates used in the test gives the highest correlation with disease activity in inflammatory bowel disease. 26 patients with Crohn's disease, 21 patients with ulcerative colitis and 27 healthy control subjects were included in the study. The patients with inflammatory bowel disease had either minimal or highly active disease or were in remission. Two disaccharides (lactulose: L, and cellobiose: C) and two smaller carbohydrates (rhamnose:R, and mannitol:M) were given orally and the urinary excretion was measured by high pressure liquid chromatography followed by pulsed amperometric electrochemical detection on a gold electrode. The ratios C/R, L/R, C/M and L/M were used as indicators for intestinal permeability. - There were no side effects of oral sugar administration. All patients tolerated the test well. Lactulose, rhamnose and cellobiose concentrations are easily be measured in the urine whereas mannitol measurement requires the use of an anion exchanger. This produced inconsistent results. Patients with Crohn's disease or with ulcerative colitis had increased permeability indices in comparison to healthy controls, even in remission. The L/R ratio gave a better differentiation between the healthy controls and patients with active disease than the other agents. Changes in disease activity are best reflected by use of cellobiose/rhamnose excretion quotient. PMID- 15546812 TI - Influence of proportional assist ventilation on diaphragmatic activity in normal subjects. AB - In six awake healthy adults we studied the physiological effects of mechanical unloading of the respiratory muscles during increased ventilatory demand. We were interested in whether respiratory muscle activity is down regulated and if this is mainly a consequence of chemical factors, i.e. CO2-reduction, or non-chemical neuromuscular inhibition. With 33 mmHg inspiratory CO2 we induced modest hyperpnea of 24.4 +/- 3.9 L/min. Proportional assist ventilation (PAV) was applied with flow-related assist of 2.5 cm H2O/L/s and volume-related assist of 6 cm H2O/L. Respiratory muscle activity was measured by transdiaphragmatic pressure. Unloading caused a 57 percent reduction of the inspiratory transdiaphragmatic pressure-time product (p<0.05), while tidal volume, breathing frequency, and breathing pattern did not significantly change. These observations suggest that during increased ventilatory requirements, PAV results in down regulation of respiratory muscle activity and that this effect is mainly a consequence of neuromechanical inhibition. PMID- 15546813 TI - Human PET/CT scanners: feasibility for oncological in vivo imaging in mice. AB - PET/CT imaging is a highly valuable oncological imaging modality. The combination of positron emission tomography (PET) and computed tomography (CT) provides the ability to accurately register molecular and metabolic aspects of cancers with anatomical and morphological findings in human clinical routine examinations and for animals in vivo research. Small animal models of mice are widely used in biomedical research for mimicking and studying the human nature, because of their genetic resemblance and the feasibility of gene transfer and gene modification. The recent generation of high performance human PET/CT scanners combines a state of the art full-ring 3D PET scanner and a high-end 16-slice CT scanner (biograph Sensation 16, Siemens AG, Erlangen, Germany). Small animals can be examined with special scanning and reconstruction protocols. The examination of tumor-bearing small animals using a modern human PET/CT revealed excellent image quality. CT can be performed with a maximum spatial resolution of 0.6 x 0.6 x 0.6 mm and PET with a maximum spatial resolution of 6.3 x 6.3 x 6.0 mm. The examination of tumor bearing small animals using human PET/CT allowed accurate correlation and evaluation of metabolic and anatomical information and is promising for in vivo research purposes. Although image quality is limited by spatial resolution, human PET/CT is widely available and expected to contribute significantly to research with small animal imaging. The investigation of cancer in small animals with PET/CT is probably one of the most challenging tasks in nuclear medicine for the evaluation of tumor growth and growth inhibition factors; development of new anti tumor drugs and measuring of anti-tumor effects; and cancer treatment response of immunotherapy, chemotherapy and radiation therapy. PMID- 15546814 TI - Early detection of diabetes retinopathy by new algorithms for automatic recognition of vascular changes. AB - Diabetes mellitus often results in diabetic retinopathy caused by pathological changes of the retinal vessel tree. Early detection of these changes can delay the disease. Image processing can reduce the workload of screeners and can play a central role in quality assurance tasks. Therefore we aimed at the refinement and development of image processing algorithms to improve the quality and cost effectiveness of screening and diagnosis of diabetic retinopathy. In order to support ophthalmologists in their routine and to enable the quantitative assessment of vascular changes in colour fundus photographs a multi-resolution approach was developed which segments the vessel tree efficiently and precisely into digital images of the retina. The vessel tracker aims at determining as correctly as possible the retinal vascular network captured on a digital image irrespective of its origin. In addition to the tracker, algorithms were developed to detect the optic disk, bright lesions such as cotton wools spots, and dark lesions such as haemorrhages. The following classification of veins and arteries identifies arteries in 78.4 % and veins in 66.5% correctly. This helps selecting conspicuous images from a great number of patients. PMID- 15546815 TI - IgM-enriched immunoglobulin (pentaglobin) positively influences the course of post-surgical intra-abdominal infections. AB - BACKGROUND: Polyvalent IgM-enriched intravenous human immunoglobulin (IVIG) preparations are discussed to be beneficial regarding sepsis outcome. MATERIALS AND METHODS: Sixty-four patients with abdominal infection were treated with Pentaglobin or Albumin. Serum levels of endotoxin and chemokines were determined. RESULTS: Incidence of fever was 19/28 in the pentaglobin and 18/26 in the albumin group, the percentage of days with fever was 34 +/- 26 for pentaglobin and 43 +/- 25 for albumin (mean +/-SD). Procalcitonin levels of the pentaglobin treated patients fell under the upper limit of normal on day six whereas levels of albumin patients remained elevated. CONCLUSION: Pentaglobin has a positive influence on the course of post-surgery intra-abdominal infection. PMID- 15546816 TI - Organ protective management of the brain-dead donor. AB - The adequate management of brain dead donors on an Intensive Care Unit (ICU) is one of the major key points for a successful transplantation of harvested organs. In addition to an invasive monitoring like in any other ICU patient these patients needs a meticulous attention to their hemodynamic. The early administration of desmopressin to treat diabetes insipidus, a differentiated use of fluid resuscitation and a distinct catecholamine support are special features of an appropriate basic treatment. The administration of corticoids has to be considered if a sufficient circulation can not be regained. PMID- 15546817 TI - Access to health care for induced abortions: analysis by means of a French national survey. AB - BACKGROUND: With an incidence of 15 per thousand, abortion is a common reproductive event in France. This study describes conditions of access to health care for abortions based on women's reports, taking into account the woman's background and the influence of the first professional contacted. METHODS: A representative sample of 2,863 women, aged 18 to 44, was interviewed by telephone between September 2000 and January 2001. Of these women, 480 had an abortion in the last 10 years. MAIN RESULTS: The choice of first professional depended on women's background, as we found differences according to age, educational level or past induced abortion. This choice affected subsequent access conditions, in terms of time delay or complexity of patterns of care to access abortion services. Women who first contacted a private gynaecologist, which is the most frequent situation in France, had more direct and shorter patterns of care. Conversely, general practitioners were associated with longer and more indirect patterns of care, especially when women were less well educated. CONCLUSION: This study reveals the heterogeneous nature of patterns of access to an abortion in France. It also raises questions concerning the training of general practitioners, who seem to be less well prepared to take charge of a request for an abortion than other professionals. Efforts must be made to better inform women and these professionals about the process required for abortions. PMID- 15546818 TI - Copper and zinc inhibit Galphas function: a nucleotide-free state of Galphas induced by Cu2+ and Zn2+. AB - The stimulatory GTP-binding protein of adenylyl cyclase (AC) regulates hormone stimulated production of cAMP. Here, we demonstrate that Cu(2+) and Zn(2+) inhibit the steady-state GTPase activity of the alpha subunit of GTP-binding protein (Galpha(s)) but do not alter its intrinsic GTPase activity. Cu(2+) and Zn(2+) decrease steady-state GTPase activity by inhibiting the binding of GTP to Galpha(s). Moreover, Cu(2+) and Zn(2+) increase GDP dissociation from Galpha(s) and render the G protein in a nucleotide-free state. However, these cations do not alter the dissociation of the guanosine 5'-3-O-(thio)triphosphate (GTPgammaS) that is already bound to the Galpha(s). Because of their ability to inhibit GTPgammaS binding, preincubation of Cu(2+) or Zn(2+) with Galpha(s) does not permit GTPgammaS to activate Galpha(s) and stimulate AC activity. However, preincubation of Galpha(s) with GTPgammaS followed by addition of Cu(2+) or Zn(2+) did not alter the ability of Galpha(s) to stimulate AC activity. Interestingly, AlF(4)(-) partially restored the ability of Galpha(s), which had been preincubated with Cu(2+) or Zn(2+), to stimulate AC; AlF(4)(-) does not permit the re-association of unbound GDP with Galpha(s). Thus, the interaction of AlF(4)(-) with the nucleotide-free Galpha(s) is sufficient to activate AC. Using antibodies to the N and C termini of Galpha(s), we show that the Cu(2+) interaction site on the G protein is in the C terminus. We conclude that Cu(2+) and Zn(2+) generate a nucleotide-free state of Galpha(s) and that, in the absence of any nucleotide, the gamma-phosphate mimic of GTP, AlF(4)(-), alters Galpha(s) structure sufficiently to permit stimulation of AC activity. Moreover, our finding that isoproterenol-stimulated AC activity was more sensitive to inhibition by Cu(2+) and Zn(2+) as compared with forskolin-stimulated activity is consistent with Galpha(s) being a primary target of these cations in regulating the signaling from receptor to AC. PMID- 15546821 TI - Role of a minimally invasive approach in the management of laparoscopic adjustable gastric banding postoperative complications. AB - HYPOTHESIS: Complications after laparoscopic adjustable gastric banding as treatment for morbid obesity may require a major reintervention. A minimally invasive approach represents an attractive management alternative for such complications. DESIGN: Prospective case series. SETTING: Major academic medical and surgical center. PATIENTS: From January 1996 to July 2003, 47 patients who had undergone laparoscopic adjustable gastric banding were operated on again. Considering the causes for reoperation, the patients were divided into 4 groups: group A had major complications (n = 26); group B, minor complications (n= 11); group C, psychological problems (n=6); and group D, insufficient weight loss (n=4). INTERVENTIONS: Forty-three procedures, 38 using general anesthesia (groups A, C, and D) and 5 using local anesthesia (group B), were performed. MAIN OUTCOME MEASURES: Feasibility, safety, and effectiveness of a minimally invasive approach in the treatment of laparoscopic adjustable gastric banding complications. RESULTS: In group A, 9 of 10 patients with irreversible gastric pouch dilatation and 15 of 16 with intragastric band migrations were treated laparoscopically. In group B, 5 ports were substituted and 2 reconnections of the catheter-port system were performed. In group C, 6 laparoscopic band removals were carried out. In group D, 4 laparoscopic revision procedures for insufficient weight loss were performed. The operative mortality was nil. The most frequent cause of reoperation was intragastric migration (37.2%). A minimally invasive approach was adopted in 94.7% of cases. CONCLUSION: Laparoscopy is safe and effective, even as a second operative procedure. PMID- 15546820 TI - Effect on oximetry of dyes used for sentinel lymph node biopsy. AB - HYPOTHESIS: There are differences between readings of peripheral blood oxygen saturation when the effect on saturation values of methylene blue is compared with that of isosulfan blue when used in sentinel lymph node biopsy in patients with breast cancer. DESIGN: Prospective randomized study. SETTING: University tertiary care center. PATIENTS: Thirty-two women undergoing surgery for breast cancer using sentinel lymph node biopsy. INTERVENTIONS: Sentinel lymph node biopsy using methylene blue (16 patients) and isosulfan blue (16 patients); there was also a control group of 6 surgical patients in whom dyes were not used. MAIN OUTCOME MEASURES: Peripheral saturation of blood using pulse oximetry, oxygen saturation by blood-gas analysis, partial oxygen pressure by blood-gas analysis, and plasma dye levels, recorded before dye injection and 15, 30, and 60 minutes afterward. RESULTS: The 2 dyes interfered with the peripheral saturation reading, but only isosulfan blue showed significant differences. The differences in blood gas analysis values between the 2 groups and with regard to the controls were not significant. CONCLUSIONS: Methylene blue interferes less than isosulfan blue in the peripheral saturation reading. Desaturation is factitious in both cases, and does not correspond to alterations in blood-gas analysis values. PMID- 15546822 TI - Screening for Depression in the Medically Ill: a comparison of self-report measures, clinician judgment, and DSM-IV diagnoses. AB - The performance of the self-report 10-item Depression in the Medically Ill scale was observed in 210 patients as part of clinical assessment by consultation liaison psychiatry clinicians. Both the Depression in the Medically Ill scale and the Beck Depression Inventory for Primary Care were completed by the patient, and the clinicians made their judgment of the presence and severity of "clinical depression" and DSM-IV affective disorder diagnoses. Both the Depression in the Medically Ill scale and the Beck Depression Inventory for Primary Care detected 85% of patients with DSM-IV major depressive episode. The Depression in the Medically Ill scale was slightly superior to the Beck Depression Inventory for Primary Care in its relationship to clinicians' judgments of clinical depression caseness. PMID- 15546823 TI - Factors affecting the relationship between the timing of psychiatric consultation and general hospital length of stay. AB - The purpose of this study was to examine the factors affecting the timing of psychiatric consultations and length of stay in the current managed care era. It also assessed the relationships between the timing of consultations and demographic/clinical characteristics. Medical records of 541 consecutive psychiatric consultation patients at a university teaching hospital in 2001 were reviewed for demographic characteristics, lengths of stay, number of days from admission to consultation, specialty services requesting consultations, reasons for the referral given by the referring physicians, and all five axes of DSM-IV. Earlier consultations independently predicted shorter lengths of stay. Delayed consultations were seen more often in women; surgical patients; those seen with a request to assess depression; and those seen with a diagnosis of adjustment disorder, delirium, or no psychiatric disorder. Delay in psychiatric consultations continues to be associated with longer lengths of stay in the current managed care environment. It is now possible that early detection strategies for high-risk patients with behavioral health problems in the medical setting, such as use of the INTERMED, may lead to reduction in delayed psychiatric consultations and thus shorter lengths of stay. PMID- 15546824 TI - The 1999 Ji-Ji (Taiwan) earthquake as a trigger for acute myocardial infarction. AB - The authors evaluated the effect of stress due to the Ji-Ji, Taiwan, earthquake, which occurred at 1:47 a.m. on September 21, 1999, on the onset of acute myocardial infarction in six counties near the earthquake epicenter. The rate of hospitalization due to acute myocardial infarction increased during the 6 weeks after the earthquake, and a significantly higher number of patients were hospitalized with acute myocardial infarction during that period, compared with the same 6-week period in the previous year (99 and 65 patients, respectively). The findings suggest that extreme emotional stress due to the natural disaster, superimposed on the stress of awakening, increased the incidence of acute myocardial infarction in this population. PMID- 15546825 TI - Diagnostic criteria for psychosomatic research and psychosocial variables in breast cancer patients. AB - The aim of the study was to examine the relationship of the Diagnostic Criteria for Psychosomatic Research (DCPR) with psychosocial variables and quality of life among cancer patients. Of 105 women with breast cancer who participated in the study, 40 (38.1%) had symptoms meeting the criteria for at least one DCPR syndrome, and 30 (28.6%) had more than one DCPR syndrome. Health anxiety, demoralization, and alexithymia were the most frequent DCPR syndromes. Patients who were diagnosed with DCPR syndromes reported higher levels of cancer-related worries and poorer quality of life than those without a DCPR diagnosis. Analysis of the single DCPR clusters and coping with cancer indicated that health anxiety was related to higher scores on the Mini-Mental Adjustment to Cancer (Mini-MAC) anxious preoccupation subscale, DCPR demoralization was related to higher scores on the Mini-MAC hopelessness subscale, and DCPR alexithymia was related to higher scores on the Mini-MAC avoidance subscale. The study indicates the usefulness of the application of the DCPR in breast cancer, although further research is needed to improve the feasibility and internal validity of DCPR constructs. PMID- 15546826 TI - Motor conversion symptoms and pseudoseizures: a comparison of clinical characteristics. AB - The authors prospectively studied consecutive neurological inpatients with either motor conversion symptoms or pseudoseizures of recent onset. Patients were administered a structured psychiatric diagnostic interview, a measure of perceived parental care, and a life events inventory. They found that patients with pseudoseizures (N=20, mean age=27 years): 1) were younger than patients with motor conversion symptoms (N=30, mean age=39 years), 2) were more likely to have a borderline personality disorder), 3) were more likely to have a lower perception of parental care and to report incest, and 4) reported more life events in the 12 months before symptom onset. These differences in their characteristics and associated factors raised the question of whether it is helpful to group patients with pseudoseizures and motor conversion symptoms in a single diagnostic category of conversion disorder. An alternative view, that gives primacy to the symptoms rather than a disorder, may enable more precise research questions to be posed. PMID- 15546827 TI - Family, health, and adolescence. AB - The present research examined the correlations between types of family relationships and adolescents' beliefs about their own health. "Healthy" adolescents (N=765) completed both the Multidimensional Health Locus of Control questionnaire and Olson's scale assessing family cohesion and adaptability. They were compared to a group of 358 adolescents diagnosed with mental disorders. Cohesion in the family of origin was a significant factor in the adolescents' feeling of control over their own health as well as in the level of power they attributed to other people. Among these adolescents, adaptability of the family of origin was positively correlated with stronger feelings of control over one's own health and with lower levels of belief in chance. Family relations were significant in the adolescents' acquisition of feelings of control over their own health. PMID- 15546828 TI - Alexithymia and somatosensory amplification in functional dyspepsia. AB - Somatosensory amplification is the tendency to report somatic sensations as intense and disturbing. Alexithymia is a personality construct characterized by difficulty recognizing emotions and a tendency to focus on external events and bodily sensations. The association of somatosensory amplification and alexithymia with functional symptoms was assessed in 111 patients with functional dyspepsia and 53 healthy comparison subjects. The subjects completed several assessment instruments, including the Somatosensory Amplification Scale and the 20-Item Toronto Alexithymia Scale. The patients with dyspepsia had modestly higher scores on measures of alexithymia (especially difficulty identifying feelings) and somatosensory amplification. Alexithymia and somatosensory amplification may play important roles in symptom generation and perception in a subset of patients with functional dyspepsia, but the importance of these constructs in this patient population appears less than previously reported. PMID- 15546829 TI - Pretransplant psychiatric and medical comorbidity of alcoholic liver disease patients who received liver transplant. AB - The authors examined the pretransplant prevalence of comorbid physical and psychological disorders in 112 alcoholic liver disease patients who received liver transplant. Fifty-six percent of the patients had comorbid hepatitis C or hepatitis B virus, 40% had used other substances in addition to alcohol, 25% met the criteria for a lifetime DSM-IV nonalcohol substance use disorder, 36% for a lifetime depressive disorder, and 12% for a lifetime anxiety disorder. The findings suggest the need for pretransplant psychiatric evaluation of alcoholic liver disease patients to identify nonalcohol substance use disorders and other psychiatric disorders that may require treatment. PMID- 15546830 TI - Antiretrovirals, Part II: focus on non-protease inhibitor antiretrovirals (NRTIs, NNRTIs, and fusion inhibitors). AB - The second in a series reviewing the HIV/AIDS antiretroviral drugs. This review summarizes the non-protease inhibitor antiretrovirals: nucleoside and nucleotide analogue reverse transcriptase inhibitors (NRTIs), the nonnucleoside reverse transcriptase inhibitors (NNRTIs), and cell membrane fusion inhibitors. In an overview format for primary care physicians and psychiatrists, this review presents the mechanism of action, side effects, toxicities, and drug interactions of these agents. PMID- 15546831 TI - Tachyarrhythmia and hypomania with horny goat weed. PMID- 15546832 TI - Confusion associated with isoniazid-induced pyridoxine deficiency. PMID- 15546835 TI - Multislice CT. PMID- 15546833 TI - Founding editor of the journal. PMID- 15546836 TI - Patient dose in multislice CT: why is it increasing and does it matter? AB - A brief review is presented of the reasons why multislice spiral/helical CT is associated with a higher radiation dose burden to the patient even than incremental CT. These include both intrinsic technological and geometric factors as well as simply a growing use of CT in an increasing number of applications. The typical magnitude of this dose burden is indicated and the basis for the anxiety that underpins it, namely the linear no-threshold (LNT) hypothesis, is discussed, together with the countervailing hypothesis that there is indeed a threshold for radiation harm in man and that the radiation doses associated with CT may lie below this threshold and may even be beneficial (radiation hormesis). There are as yet no certainties in this important area but it is argued that it is not a given that the doses associated with CT are harmful. PMID- 15546837 TI - Multidetector CT: what do we do with all the images generated? AB - Multidetector spiral CT technology generates much more acquisition data than the previous generation of single detector spiral CT machines, and permits more sophisticated and better quality post-processing options, which in turn generate even more imaging data. Such large quantities of data have repercussions upon the mode of image interpretation (hard copy vs soft copy), the speed of data transmission and the storage of these data. PMID- 15546838 TI - Scanning protocols for multislice CT scanners. AB - The new multislice CT (MSCT) technology enables examinations to be completed with much greater flexibility in terms of shorter times and improved z-axis resolution. For this to be achieved, a very large amount of data may be produced, with implications on the way the data are handled and reviewed. In addition, there is the issue of radiation dose to the patient and the effect that multislice scanning has on it. In this paper, the basic physics principles of MSCT are reviewed and clarification is given why and where these differ from single slice scanning. Examples of scanning protocols are given with reference to patient dose. These protocols vary depending on the make and model of the scanner and the type of examination. PMID- 15546839 TI - Brain imaging using multislice CT: a personal perspective. AB - This article outlines the use of multislice CT of the brain with regard to routine scanning, as well as high speed and/or high resolution multiplanar reconstructions. It describes in detail the recent developments in cerebral perfusion scanning and CT arteriography. It will also outline the approach of the University Hospital of Wales to imaging with this new technology, emphasizing the differences between helical and non-helical techniques. Most of the images are taken from 16-slice machines, but the methods described are applicable to 4- and 8-slice scanners, and can readily be modified for the forthcoming 64-detector array equipment that will be available within the next few months. PMID- 15546840 TI - Multislice CT angiography: a practical guide to CT angiography in vascular imaging and intervention. AB - The development of multidetector CT (MDCT) has revolutionized CT angiography (CTA). Not only are new techniques now in the remit of CTA, but all the studies previously performed on single slice or helical CT can now be done with better resolution. The advantage of MDCT relevant to CTA is the ability to acquire high resolution, near isotropic data sets in a shorter acquisition time. Also important is the ability to achieve a longer scanning range in the arterial phase, which has seen the introduction of CTA of the peripheral arterial system. Image processing techniques have also progressed rapidly, with simplification of a previously cumbersome process. The high spatial resolution and relatively non invasive nature make MDCT angiography a strong and serious competitor to established vascular imaging techniques. The implication is that traditional diagnostic pathways for evaluation of the vascular system have changed. PMID- 15546841 TI - Multislice CT pulmonary angiography and CT venography. AB - Pulmonary embolism (PE) is a common and potentially lethal complication of deep venous thrombosis. Clinical diagnosis is difficult and treatment carries significant potential side effects. High sensitivity and specificity of any diagnostic modality for PE are desirable. Helical CT pulmonary angiography (CTPA) offers this by direct visualization of embolic material within the pulmonary arteries. For a number of reasons, including the mobility and orientation of the pulmonary vessels and the peripheral nature of some emboli, the accuracy of CTPA is directly related to the spatial resolution of the technique used. The advent of multislice technology lends itself to improved spatial and temporal resolution and has led to a significant improvement on earlier CTPA results. Indirect CT venography may also be performed as an adjunct to CTPA to help demonstrate culprit thrombus within the lower limbs. PMID- 15546842 TI - Multislice CT in imaging of trauma of the spine, pelvis and complex foot injuries. AB - Multislice CT (MSCT) has greatly enhanced the performance of CT scanners and has vastly improved imaging of musculoskeletal trauma. Fast, high resolution scanning is now possible. In our institution, MSCT is an essential part of the imaging of the traumatized patient. The advantages of volume imaging, such as multiplanar reconstructions (MPRs) with near isotropic viewing, three-dimensional imaging and thick slice (wedge) MPRs (mimicking conventional radiographs), enable more accurate assessment of complex anatomical areas such as the spine, pelvis and foot. We discuss the general principles of scanning for musculoskeletal trauma and describe our experience of MSCT of the traumatized spine, pelvis and foot. PMID- 15546843 TI - Multislice CT of the abdomen. AB - Major advances have been made in CT technology since its introduction 30 years ago. Examination time has been markedly reduced and it is possible to cover considerably larger scan ranges. Multidetector technology now allows near isotopic imaging with the ability to perform high quality multiplanar reconstructions. It is also now possible to image in well defined vascular phases. In this article we share our experience in the use of multislice CT and detail protocols used in imaging of the abdomen. PMID- 15546844 TI - Multislice CT urography: state of the art. AB - Recent improvements in helical CT hardware and software have provided imagers with the tools to obtain an increasingly large number of very thin axial images. As a result, a number of new applications for multislice CT have recently been developed, one of which is CT urography. The motivation for performing CT urography is the desire to create a single imaging test that can completely assess the kidneys and urinary tract for urolithiasis, renal masses and mucosal abnormalities of the renal collecting system, ureters and bladder. Although the preferred technique for performing multislice CT urography has not yet been determined and results are preliminary, early indications suggest that this examination can detect even subtle benign and malignant urothelial abnormalities and that it has the potential to completely replace excretory urography within the next several years. An important limitation of multislice CT urography is increased patient radiation exposure encountered when some of the more thorough recommended techniques are utilized. PMID- 15546845 TI - Multislice CT in cardiac and coronary angiography. AB - In the last 2 years, mechanical multidetector-row CT (MDCT) systems with simultaneous acquisition of four slices and a half second scanner rotation time have become widely available. Data acquisition with these scanners allows for considerably faster coverage of the heart volume compared with single slice scanning. This increased scan speed can be used for retrospective gating together with 1 mm collimated slice widths and allows coverage of the entire cardiac volume in one breath-hold. First results from studies in correlation with intracoronary ultrasound suggest that MDCT technology not only offers the possibility to visualize intracoronary stenoses non-invasively, but also to differentiate plaque morphology. This is especially the case with the next generation of 16-row MDCT systems. An increased number of simultaneously acquired slices and submillimetre collimation for cardiac applications allows true isotropic scanning with high temporal resolution. Contrast-enhanced MDCT is a promising non-invasive technique for the detection, visualization and characterization of stenotic artery disease. It could act as a gatekeeper prior to cardiac catherization and finally replace conventional diagnostic modalities. PMID- 15546846 TI - High resolution volume imaging of airways and lung parenchyma with multislice CT. AB - The value of multislice CT (MSCT) in imaging the peripheral airways and lung parenchyma has not been widely investigated. In this article the authors' experience in the use of MSCT (4-slice scanner) in imaging patients with suspected parenchymal lung disease or airways abnormalities will be presented. The technique described should be modified with the more modern 8-slice or 16 slice scanners. The whole thorax is scanned contiguously using 4 x 1 mm collimation from the lung bases up to apices in end-inspiration while the patient is in the prone position. Collimation of 2 x 0.5 mm is used at 8-10 levels evenly spaced in expiratory scans and also in the breathless patient who is scanned during gentle breathing. High resolution images of the lungs (1 mm slice thickness) are reconstructed in the following planes: axial (10 mm apart from apices to bases), coronal (six evenly spaced through the chest) and sagittal (four images evenly spaced through each lung). Paddlewheel reconstruction is used if further assessment of the airways is required, and three-dimensional imaging is used mainly for assessment of the trachea and major bronchi. Contiguous axial images (10 mm slice thickness) of the whole lung and mediastinum are also produced and referred to as a screenogram. Axial images of 1 mm slice thickness are produced with expiratory scans and for breathless patients. All the images are produced independently by the radiographic staff and are provided as hard copies (20 frames/film) for reporting. However, if facilities are adequate, direct reporting from the workstation is more effective in reviewing large number of images. The technique is effective in assessment of infiltrative lung disease, emphysema, bronchiectasis and central airways. The screenogram offers comprehensive evaluation of the lung and mediastinum, but the radiation dose associated with high resolution volume imaging remains a source of concern. PMID- 15546848 TI - Imaging takes a quantum leap. AB - Semiconducting nanocrystals, or quantum dots (QDs), have emerged as a new tool in physiological imaging, combining high brilliance, photostability, broad excitation but very narrow emission spectra, and surface chemistry compatible with biomolecular conjugation. In this review, we demonstrate the power of QDs in diverse applications, including long-term in vivo fluorescence imaging. PMID- 15546849 TI - The first nobel prize for integrated systems physiology: Ivan Petrovich Pavlov, 1904. PMID- 15546850 TI - The molecular physiology of tight junction pores. AB - Tight junctions form selective barriers that regulate paracellular transport across epithelia. A large family of tetraspanning cell-cell adhesion proteins called claudins create the barrier and regulate electrical resistance, size, and ionic charge selectivity. Study of inherited human claudin diseases and the outcome of the genetic manupulation of claudins in mice, Drosophila, and Caenorhabditis elegans are furthering our understanding of paracellular physiology. PMID- 15546851 TI - Implications of gene networks for understanding resilience and vulnerability in the kidney branching program. AB - Branching morphogenesis in the kidney is tightly regulated. Whereas disruption of certain pathways produces catastrophic effects, numerous instances exist in which mutation of ostensibly key molecules has minimal apparent phenotypic consequence. We suggest how the network structure of gene interactions in the branching program might explain these findings as well as apparant discrepancies between in vivo and in vitro studies. Emerging genetic, cell-biological, and microarray data should help test and/or clarify these ideas. PMID- 15546852 TI - Common cues in vascular and axon guidance. AB - Blood vessels and nerves are structured in architecturally similar organ systems and show functional relationships. Indeed, vascular and neuronal cells are guided in their journey throughout the body by the same attractive and repulsive factors that respectively activate and inhibit the function of integrin-adhesive receptors. PMID- 15546853 TI - ARC channels: a novel pathway for receptor-activated calcium entry. AB - In many nonexcitable cells, stimulation with low agonist concentrations specifically activates Ca2+ entry via arachidonic acid-regulated, highly Ca2+ selective ARC channels. Only at high agonist concentrations are the more widely studied store-operated channels activated, producing sustained elevated cytosolic Ca2+ concentration signals. These signals activate calcineurin, which in turn inhibits the ARC channels, resulting in a "reciprocal regulation" of these two distinct Ca2+-entry pathways that may have important functional implications for the cell. PMID- 15546854 TI - Regulation of ion transport by the NHERF family of PDZ proteins. AB - NHERFs are the best-studied apical PDZ proteins that are highly expressed in epithelial cells. Molecular and cellular studies over the past decade show that NHERFs regulate the targeting or trafficking of ion transporters and other membrane proteins and transduce physiological and pathophysiological signals that regulate ion homeostasis in mammals. PMID- 15546855 TI - Surprising versatility of Na+-glucose cotransporters: SLC5. AB - SLC5 is an ancient gene family with 11 members in the human genome. These membrane proteins have diverse, multiple functions ranging from actively transporting solutes, ions, and water, to channeling water and urea, to sensing glucose in cholinergic neurons. Metabolic disorders have been identified that are associated with congenital mutations in two of the human genes. PMID- 15546856 TI - Recent insights into the structure and mechanism of the sodium pump. AB - The sodium pump (or Na-K-ATPase) is essential to the function of animal cells. Publication of the related calcium pump (SERCA) structure together with several recent results from a variety of approaches allow us to propose a mechanistic model to answer the question: "How does the sodium pump pump?" PMID- 15546857 TI - Reduced expression of the insulin receptor in mouse insulinoma (MIN6) cells reveals multiple roles of insulin signaling in gene expression, proliferation, insulin content, and secretion. AB - The role of insulin signaling in pancreatic beta cells has become increasingly apparent. Stably transformed insulinoma cell lines (MIN6) were created with small interfering RNA resulting in the reduction of insulin receptor (IR) expression up to 80% (insulin receptor knockdown, IRKDDelta80). Functionally perturbed IR signaling was confirmed with the absence of insulin-stimulated insulin receptor substrate 1 tyrosine phosphorylation. Additionally, Akt phosphorylation was reduced and responded poorly to glucose stimulation. Gene expression profiling revealed that reduced IR expression was associated with alterations in expression of >1,500 genes with diverse functions. IRKD cells exhibited low rate of proliferation due to delay in transition from G0/G1 to S phase, whereas susceptibility to apoptosis did not differ from that of control cells. Insulin content was reduced in proportion to the reduction of IR. IRKD cells maintained glucose responsiveness as measured by NADPH generation, whereas Ca2+ responses and insulin secretion were enhanced. IRKDDelta80 and control cells were treated with glucose (25 mm) or insulin (100 nm) for 45 min, and gene expression profiles were assessed. Transcriptional activation of several hundred early response genes common to both glucose and insulin stimulation was observed in control cells. In IRKDDelta80 cells, insulin failed to activate any genes as anticipated. Importantly, glucose stimulation of gene expression in IRKDDelta80 cells showed that most genes previously activated by glucose were no longer activated, suggesting a major autocrine/paracrine effect of insulin on glucose-regulated gene expression. On the other hand, there were a number of glucose-regulated genes in the IRKDDelta80 cells that were not previously observed in control cells, suggesting a feedback regulation of insulin signaling on glucose-regulated gene expression. These results demonstrate important roles of the insulin receptor in islet beta cell gene expression and function and may serve to elucidate molecular defects in animal models with diminished beta cell insulin signaling. PMID- 15546858 TI - Activation of ataxia telangiectasia mutated by DNA strand break-inducing agents correlates closely with the number of DNA double strand breaks. AB - The protein kinase ataxia telangiectasia mutated (ATM) is activated when cells are exposed to ionizing radiation (IR). It has been assumed that ATM is specifically activated by the few induced DNA double strand breaks (DSBs), although little direct evidence for this assumption has been presented. DSBs constitute only a few percent of the IR-induced DNA damage, whereas the more frequent single strand DNA breaks (SSBs) and base damage account for over 98% of the overall DNA damage. It is therefore unclear whether DSBs are the only IR induced DNA lesions that activate ATM. To test directly whether or not DSBs are responsible for ATM activation, we exposed cells to drugs and radiation that produce different numbers of DSBs and SSBs. We determined the resulting ATM activation by measuring the amount of phosphorylated Chk2 and the numbers of SSBs and DSBs in the same cells after short incubation periods. We found a strong correlation between the number of DSBs and ATM activation but no correlation with the number of SSBs. In fact, hydrogen peroxide, which, similar to IR, induces DNA damage through hydroxyl radicals but fails to induce DSBs, did not activate ATM. In contrast, we found that calicheamicin-induced strand breaks activated ATM more efficiently than IR and that ATM activation correlated with the relative DSB induction by these agents. Our data indicate that ATM is specifically activated by IR-induced DSBs, with little or no contribution from SSBs and other types of DNA damage. These findings have implications for how ATM might recognize DSBs in cells. PMID- 15546859 TI - In vitro characterization of a tRNA editing activity in the mitochondria of Spizellomyces punctatus, a Chytridiomycete fungus. AB - In the chytridiomycete fungus, Spizellomyces punctatus, all eight of the mitochondrially encoded tRNAs are predicted to have one or more base pair mismatches at the first three positions of their aminoacyl acceptor stems. These tRNAs are edited post-transcriptionally by replacement of the 5'-nucleotide in each mismatched pair with a nucleotide that can form a standard Watson-Crick base pair with its counterpart in the 3'-half of the stem. The type of mitochondrial tRNA editing found in S. punctatus also occurs in Acanthamoeba castellanii, a distantly related amoeboid protist. Using an S. punctatus mitochondrial extract, we have developed an in vitro assay of tRNA editing in which nucleotides are incorporated into various tRNA substrates. Experiments employing synthetic transcripts revealed that the S. punctatus tRNA editing activity incorporates nucleotides on the 5'-side of substrate tRNAs, uses the 3'-sequence as a template for incorporation, and adds nucleotides in a 3'-to-5' direction. This activity can add nucleotides to a triphosphorylated 5'-end in the absence of ATP but requires ATP to add nucleotides to a monophosphorylated 5'-end; moreover, it functions independently of the state of tRNA 3' processing. These data parallel results obtained in a previous in vitro study of A. castellanii tRNA editing, suggesting that remarkably similar activities function in the mitochondria of these two organisms. The evolutionary origins of these activities are discussed. PMID- 15546860 TI - Prion-associated increases in Src-family kinases. AB - The prion diseases result from the generation and propagation of an abnormal conformer of the prion protein. It is unclear how this molecular event disrupts neuronal function and viability. Current evidence argues it is not due to loss of normal prion protein activity or direct toxic effects of the abnormal conformer. Both the normal and abnormal prion proteins are glycosylphosphatidylinositol linked membrane proteins. Conversion to the abnormal isoform results in the formation and accumulation of prion protein aggregates. Because aggregation of glycosylphosphatidylinositol-linked proteins activates Src-family kinases, the activation status and levels of the Src-family kinases in prion disease were investigated. Elevations of Src-family kinases were found in a cell culture model and two separate animal models of prion disease. The elevations in Src kinases preceded the onset of symptoms and occurred concurrently with the appearance of detergent-insoluble prion protein. In addition, the total level of kinases phosphorylated at tyrosine residues associated with activation was increased. Similar alterations were not present in brain homogenates from presymptomatic animals early in the disease course, prion protein-ablated animals, or end-stage Tg2576 mice overexpressing mutant amyloid precursor protein. Identification of similar elevations in cell culture and animal model systems suggests the elevations are a specific response to the presence of the disease-associated conformer. Abnormal regulation of these signal transduction cascades may be a key element in the cellular pathology of the prion diseases. PMID- 15546861 TI - Dephosphorylation of tau by protein phosphatase 5: impairment in Alzheimer's disease. AB - Protein phosphatase (PP) 5 is highly expressed in the mammalian brain, but few physiological substrates have yet been identified. Here, we investigated the kinetics of dephosphoryation of phospho-tau by PP5 and found that PP5 had a K(m) of 8-13 microm toward tau, which is similar to that of PP2A, the major known tau phosphatase. This K(m) value is within the range of intraneuronal tau concentration in human brain, suggesting that tau could be a physiological substrate of both PP5 and PP2A. PP5 dephosphorylated tau at all 12 Alzheimer's disease (AD)-associated abnormal phosphorylation sites studied, with different efficiency toward each site. Thr(205), Thr(212), and Ser(409) of tau were the most favorable sites; Ser(199), Ser(202), Ser(214), Ser(396), and Ser(404) were less favorable sites; and Ser(262) was the poorest site for PP5. Overexpression of PP5 in PC12 cells resulted in dephosphorylation of tau at multiple phosphorylation sites. The activity but not the protein level of PP5 was found to be decreased by approximately 20% in AD neocortex. These results suggest that tau is probably a physiological substrate of PP5 and that the abnormal hyperphosphorylation of tau in AD might result in part from the decreased PP5 activity in the diseased brains. PMID- 15546862 TI - Molecular characterization of major cat allergen Fel d 1: expression of heterodimer by use of a baculovirus expression system. AB - Fel d 1 is a major cat allergen inducing allergic rhinitis and asthma in sensitized individuals. It has a more complex structure when compared with other allergens and therefore expression of recombinant Fel d 1 has been considered a challenge. The present study shows for the first time that a Baculovirus expression system is able to produce an intact rFel d 1 molecule that is glycosylated and structurally equivalent to the natural cat allergen, nFel d 1. Enzymatic digestion of rFel d 1 and further analysis by use of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) resulted in a complete coverage of the amino acid sequence of rFel d 1. In addition, the three disulfide bridges at the positions alpha70-beta7, alpha44 beta48, and alpha3-beta73 were verified. The N-glycan structure of rFel d 1 was investigated by a combination of MALDI-TOF MS and monosaccharide analysis by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAC). The N-glycosylation analyses of rFel d 1 refer to a pattern of glycoforms including core alpha1.3-fucosylation that is different from nFel d 1. Further characterization by use of human serum IgE, histamine release, and lymphocyte proliferation assays demonstrated that the immunological characteristics of rFel d 1 are similar to those of nFel d 1. Detailed characterization of both natural and recombinant allergens provides tools to explore immunological mechanisms associated with allergen sensitization and desensitization. PMID- 15546863 TI - Full activation of PKB/Akt in response to insulin or ionizing radiation is mediated through ATM. AB - The gene mutated in ataxia telangiectasia, ATM, has been implicated in several cell functions such as cell cycle control and response to DNA damage and insulin. PKB/Akt has also been implicated in the cellular response to insulin, gamma radiation, and cell cycle control. Interestingly, lack of PKB/Akt function in vivo is able to mimic some phenotypic abnormalities associated with ataxia telangiectasia (AT). Here we show that ATM is a major determinant of full PKB/Akt activation in response to insulin or gamma-radiation. This effect is mediated through the phosphatidylinositol 3-kinase domain of ATM that specifically affects Akt serine 473 phosphorylation. This conclusion was inferred from the results obtained in transient transfection assays using exogenous PKB/Akt and ATM in Cos cells. Moreover, the use of ATM inhibitors or small interfering RNA confirmed our observation. Further supporting these results, we also observed that biological responses tightly regulated by Akt, such as transcription factor of the forkhead family activity after insulin treatment or gamma-radiation response, were altered in cell lines derived from AT patients and knockout mice for ATM in which phosphorylation in serine 473 was almost abolished. This study proposes new clues in the search of the unknown PDK2 and new explanations for the radiosensitivity or insulin intolerance described more than 30 years ago in AT patients. PMID- 15546864 TI - Regulation of cystic fibrosis transmembrane regulator trafficking and protein expression by a Rho family small GTPase TC10. AB - The cystic fibrosis transmembrane conductance regulator (CFTR)-interacting protein, CFTR-associated ligand (CAL) down-regulates total and cell surface CFTR by targeting CFTR for degradation in the lysosome. Here, we report that a Rho family small GTPase TC10 interacts with CAL. This interaction specifically up regulates CFTR protein expression. Co-expression of the constitutively active form, TC10Q75L, increases total and cell surface CFTR in a dose-dependent fashion. Moreover, co-expression of the dominant-negative mutant TC10T31N causes a dose-dependent reduction in mature CFTR. The effect of TC10 is independent of the level of CFTR expression, because a similar effect was observed in a stable cell line that expresses one-tenth of CFTR. Co-expression of TC10Q75L did not have a similar effect on the expression of plasma membrane proteins such as Frizzled-3 and Pr-cadherin or cytosolic proteins such as tubulin and green fluorescent protein. TC10Q75L also did not have a similar effect on the vesicular stomatitis virus glycoprotein. Co-expression of constitutively active and dominant-negative forms of Cdc42 or RhoA did not affect CFTR expression in a manner similar to TC10, indicating that the effect of TC10 is unique within the Rho family. Metabolic pulse-chase experiments show that TC10 did not affect CFTR maturation, suggesting that it exerts its effects on the mature CFTR. Importantly, TC10Q75L reverses CAL-mediated CFTR degradation, suggesting that TC10Q75L inhibits CAL-mediated degradation of CFTR. TC10Q75L does not operate by reducing CAL protein expression or its ability to form dimers or interact with CFTR. Interestingly, the expression of TC10Q75L causes a dramatic redistribution of CAL from the juxtanuclear region to the plasma membrane where the two molecules overlap. These data suggest that TC10 regulates both total and plasma membrane CFTR expression by interacting with CAL. The GTP-bound form of TC10 directs the trafficking of CFTR from the juxtanuclear region to the secretory pathway toward the plasma membrane, away from CAL-mediated degradation of CFTR in the lysosome. PMID- 15546865 TI - Acquisition of unprecedented phosphatidylinositol 3,5-bisphosphate rise in hyperosmotically stressed 3T3-L1 adipocytes, mediated by ArPIKfyve-PIKfyve pathway. AB - Unlike yeast, where hyperosmotic stress induces a dramatic increase in phosphatidylinositol 3,5-bisphosphate (PtdIns 3,5-P(2)) synthesis, in mammalian cells, although activating a complex array of signaling events, hyperosmotic stress fails to up-regulate PtdIns 3,5-P(2), indicating the PtdIns 3,5-P(2) pathway is not involved in mammalian osmo-protective responses. Here we report an unexpected and marked PtdIns 3,5-P(2) increase in response to hyperosmotic stress in differentiated 3T3-L1 adipocytes. Because this effect was not observed in the precursor preadipocytes, a specific role during acquisition of the adipocyte phenotype and transition into insulin-responsive cells could be suggested. However, acute insulin action did not result in a measurable PtdIns 3,5-P(2) rise, indicating the PtdIns 3,5-P(2) pathway is a specific hyperosmotically activated signaling cascade selectively operating in differentiated 3T3-L1 adipocytes. Hyperosmolarity activates different components of several kinase cascades, including p38 mitogen-activated protein and tyrosine kinases, but these appear to be separate from the activated PtdIns 3,5-P(2) pathway. Because PtdIns 3,5-P(2) is primarily produced by PIKfyve-catalyzed synthesis and requires the upstream activator hVac14 (called herein ArPIKfyve) that physically associates with and activates PIKfyve, we examined the contribution of ArPIKfyve-PIKfyve for the hyperosmotic stress-induced rise in PtdIns 3,5-P(2). Small interfering RNA directed gene silencing to selectively deplete ArPIKfyve or PIKfyve in 3T3-L1 adipocytes determined the ArPIKfyve-PIKfyve axis fully accountable for the hyperosmotically activated PtdIns 3,5-P(2). Together these results reveal a previously uncharacterized PtdIns 3,5-P(2) pathway activated selectively in hyperosmotically stressed 3T3-L1 adipocytes and suggest a plausible role for PtdIns 3,5-P(2) in the osmo-protective response mechanism in this cell type. PMID- 15546866 TI - Alternative splicing microarrays reveal functional expression of neuron-specific regulators in Hodgkin lymphoma cells. AB - Alternative splicing provides a versatile mechanism of gene regulation, which is often subverted in disease. We have used customized oligonucleotide microarrays to interrogate simultaneously the levels of expression of splicing factors and the patterns of alternative splicing of genes involved in tumor progression. Analysis of RNAs isolated from cell lines derived from Hodgkin lymphoma tumors indicate that the relative abundance of alternatively spliced isoforms correlates with transformation and tumor grade. Changes in expression of regulators were also detected, and a subset sample was confirmed at the protein level. Ectopic expression of neuron-specific splicing regulatory proteins of the Nova family was observed in some cell lines and tumor samples, correlating with expression of a neuron-specific mRNA isoform of JNK2 kinase. This microarray design can help assess the role of alternative splicing in a variety of biological and medical problems and potentially serve as a diagnostic tool. PMID- 15546867 TI - Transforming growth factor-beta (TGF-beta) type I receptor/ALK5-dependent activation of the GADD45beta gene mediates the induction of biglycan expression by TGF-beta. AB - We have recently shown that induction of biglycan (BGN) expression by transforming growth factor-beta1 (TGF-beta1) required sequential activation of both Smad and p38 mitogen-activated protein kinase signaling (Ungefroren, H., Lenschow, W., Chen, W.-B., and Kalthoff, H. (2003) J. Biol. Chem. 278, 11041 11049). Here, we have analyzed the receptors through which TGF-beta1 controls expression of BGN and GADD45beta, the latter of which is postulated to link early Smad signaling to delayed activation of p38. Ectopic expression of a dominant negative mutant of the TGF-beta type II receptor in PANC-1 cells abrogated TGF beta-induced BGN up-regulation. Similarly, inhibition of the TGF-beta type I receptor/ALK5 with either SB431542 or by enforced stable expression of a kinase dead mutant greatly attenuated the TGF-beta effect on both BGN and GADD45beta expression in PANC-1 and MG-63 cells. The enhancing effect of ALK5 on TGF-beta mediated GADD45beta and BGN expression and on GADD45beta promoter activity was also dependent on its ability to activate Smad signaling, because an ALK5 mutant defective in Smad activation (TbetaRImL45) but with an otherwise functional kinase domain failed to mediate these responses. The TGF-beta/ALK5 effect on p38 activation and BGN expression was mimicked by overexpression of GADD45beta alone (in the absence of TGF-beta stimulation) and suppressed upon antisense inhibition of GADD45beta expression. These results show that TGF-beta induces BGN expression through (the Smad-activating function of) ALK5 and GADD45beta and suggest that the sensitivity of MyD118 to activation by TGF-beta, which varies between tissues, ultimately determines the strength of the TGF-beta effect on BGN. PMID- 15546868 TI - Mirk/dyrk1B decreases the nuclear accumulation of class II histone deacetylases during skeletal muscle differentiation. AB - Mirk/dyrk1B is a member of the dyrk/minibrain family of serine/threonine kinases that mediate the transition from growth to differentiation in lower eukaryotes and mammals. Depletion of endogenous Mirk from C2C12 myoblasts by RNA interference blocks skeletal muscle differentiation (Deng, X., Ewton, D., Pawlikowski, B., Maimone, M., and Friedman, E. (2003) J. Biol. Chem. 278, 41347 41354). We now demonstrate that knockdown of Mirk blocks transcription of the muscle regulatory factor myogenin. Co-expression of Mirk with MEF2C, but not MyoD or Myf5, enhanced activation of the myogenin promoter in a Mirk kinase-dependent manner. Mirk activated MEF2 not through direct phosphorylation of MEF2 but by phosphorylation of its inhibitors, the class II histone deacetylases (HDACs). MEF2 is sequestered by class II HDACs such as HDAC5 and MEF2-interacting transcriptional repressor (MITR). Mirk antagonized the inhibition of MEF2C by MITR, whereas kinase-inactive Mirk was ineffective. Mirk phosphorylates class II HDACs at a conserved site within the nuclear localization region, reducing their nuclear accumulation in a dose-dependent and kinase-dependent manner. Moreover, less mutant MITR phosphomimetic at the Mirk phosphorylation site localized in the nucleus than wild-type MITR. Regulation of class II HDACs occurs by multiple mechanisms. Others have shown that calcium signaling leads to phosphorylation of HDACs at 14-3-3-binding sites, blocking their association with MEF2 within the nucleus. Mirk provides another level of regulation. Mirk is induced within the initial 24 h of myogenic differentiation and enables MEF2 to transcribe the myogenin gene by decreasing the nuclear accumulation of class II HDACs. PMID- 15546869 TI - Roles of conserved proline and glycosyltransferase motifs of EmbC in biosynthesis of lipoarabinomannan. AB - D-Arabinans, composed of D-arabinofuranose (D-Araf), dominate the structure of mycobacterial cell walls in two settings, as part of lipoarabinomannan (LAM) and arabinogalactan, each with markedly different structures and functions. Little is known of the complexity of their biosynthesis. beta-D-Arabinofuranosyl-1 monophosphoryldecaprenol is the only known sugar donor. EmbA, EmbB, and EmbC, products of the paralogous genes embA, embB, and embC, the sites of resistance to the anti-tuberculosis drug ethambutol (EMB), are the only known implicated enzymes. EmbA and -B apparently contribute to the synthesis of arabinogalactan, whereas EmbC is reserved for the synthesis of LAM. The Emb proteins show no overall similarity to any known proteins beyond Mycobacterium and related genera. However, functional motifs, equivalent to a proline-rich motif of several bacterial polysaccharide co-polymerases and a superfamily of glycosyltransferases, were found. Site-directed mutagenesis in glycosyltransferase superfamily C resulted in complete ablation of LAM synthesis. Point mutations in three amino acids of the proline motif of EmbC resulted in marked reduction of LAM-arabinan synthesis and accumulation of an unknown intermediate and of the known precursor lipomannan. Yet the pattern of the differently linked d-Araf units observed in wild type LAM-arabinan was largely retained in the proline motif mutants. The results allow for the presentation of a unique model of arabinan synthesis. PMID- 15546870 TI - New genes implicated in the protection of anaerobically grown Escherichia coli against nitric oxide. AB - Nitric oxide produced by activated macrophages plays a key role as one of the immune system's weapons against pathogens. Because the lifetime of nitric oxide is short in aerobic conditions, whereas in anaerobic conditions the cytotoxic effects of nitric oxide are greatly increased as in the infection/inflammation processes, it is important to establish which systems are able to detoxify nitric oxide under anaerobic conditions. In the present work a new set of Escherichia coli K-12 genes conferring anaerobic resistance to nitric oxide is presented, namely the gene product of YtfE and a potential transcriptional regulator of the helix-turn-helix LysR-type (YidZ). The crucial role of flavohemoglobin for anaerobic nitric oxide protection is also demonstrated. Furthermore, nitric oxide is shown to cause a significant alteration of the global E. coli gene transcription profile that includes the increase of the transcript level of genes encoding for detoxification enzymes, iron-sulfur cluster assembly systems, DNA repairing enzymes, and stress response regulators. PMID- 15546871 TI - Identification of transcriptional networks during liver regeneration. AB - The molecular analysis of mammalian cellular proliferation in vivo is limited in most organ systems by the low turnover and/or the asynchronous nature of cell cycle progression. A notable exception is the partial hepatectomy model, in which quiescent hepatocytes reenter the cell cycle and progress in a synchronous fashion. Here we have exploited this model to identify regulatory networks operative in the mammalian cell cycle. We performed microarray-based expression profiling on livers 0-40 h post-hepatectomy corresponding to G0, G1, and S phases. Differentially expressed genes were identified using the statistical analysis program PaGE (Patterns from Gene Expression), which was highly accurate as confirmed by quantitative reverse transcription-PCR of randomly selected targets. A shift in the transcriptional program from genes involved in lipid and hormone biosynthesis in the quiescent liver to those contributing to cytoskeleton assembly and DNA synthesis in the proliferating liver was demonstrated by biological theme analysis. In a novel approach, we employed computational pathway analysis tools to identify specific regulatory networks operative at various stages of the cell cycle. This allowed us to identify a large cluster of genes controlling mitotic spindle assembly and checkpoint control at the 40-h time point as regulated at the mRNA level in vivo. PMID- 15546872 TI - MBNL1 is the primary determinant of focus formation and aberrant insulin receptor splicing in DM1. AB - In myotonic dystrophy 1 (DM1), aggregation of the mutant DMPK RNA into RNA protein complexes containing MBNL1 and MBNL2 has been linked to aberrant splicing of the insulin receptor (IR) RNA. In a parallel line of investigation, elevated levels of CUG-binding protein (CUG-BP) have been shown to result in altered IR splicing in DM1. The relative importance of MBNL1, MBNL2, and CUG-BP in DM1 pathogenesis is, however, unclear. Here we have demonstrated that either small interfering RNA-mediated down-regulation of MBNL1 and MBNL2 or the overexpression of CUG-BP in normal myoblasts results in abnormal IR splicing. Our results suggest that CUG-BP regulates the equilibrium of splice site selection by antagonizing the facilitatory activity of MBNL1 and MBNL2 on IR exon 11 splicing in a dose-dependent manner. We have shown that CUG-BP levels are elevated in DM1 cells by mechanisms that are independent of MBNL1 and MBNL2 loss. Importantly, rescue experiments in DM1 myoblasts demonstrated that loss of MBNL1 function is the key event, whereas the overexpression of CUG-BP plays a secondary role in the aberrant alternative splicing of IR RNA in DM1. Small interfering RNA-mediated down-regulation of MBNL1, MBNL2, and CUG-BP in DM1 myoblasts demonstrated that MBNL1 plays a critical role in the maintenance of DM1 focus integrity. Thus, these experiments demonstrate that sequestration of MBNL1 by the expanded CUG repeats is the primary determinant of both DM1 focus formation and the abnormal splicing of the IR RNA in DM1 myoblasts. The data therefore support MBNL1 mediated therapy for DM1. PMID- 15546873 TI - Endoplasmic reticulum stress stimulates heme oxygenase-1 gene expression in vascular smooth muscle. Role in cell survival. AB - Heme oxygenase-1 (HO-1) is a cytoprotective protein that catalyzes the degradation of heme to biliverdin, iron, and carbon monoxide (CO). In the present study, we found that endoplasmic reticulum (ER) stress induced by a variety of experimental agents stimulated a time- and concentration-dependent increase in HO 1 mRNA and protein in vascular smooth muscle cells (SMC). The induction of HO-1 by ER stress was blocked by actinomycin D or cycloheximide and was independent of any changes in HO-1 mRNA stability. Luciferase reporter assays indicated that ER stress stimulated HO-1 promoter activity via the antioxidant response element. Moreover, ER stress induced the nuclear import of Nrf2 and the binding of Nrf2 to the HO-1 antioxidant response element. Interestingly, ER stress stimulated SMC apoptosis, as demonstrated by annexin V binding, caspase-3 activation, and DNA laddering. The induction of apoptosis by ER stress was potentiated by HO inhibition, whereas it was prevented by addition of HO substrate. In addition, exposure of SMC to exogenously administered CO inhibited ER stress-mediated apoptosis, and this was associated with a decrease in the expression of the proapoptotic protein, GADD153. In contrast, the other HO-1 products failed to block apoptosis or GADD153 expression during ER stress. These results demonstrated that ER stress is an inducer of HO-1 gene expression in vascular SMC and that HO-1-derived CO acts in an autocrine fashion to inhibit SMC apoptosis. The capacity of ER stress to stimulate the HO-1/CO system provides a novel mechanism by which this organelle regulates cell survival. PMID- 15546874 TI - Galectin-4 binds to sulfated glycosphingolipids and carcinoembryonic antigen in patches on the cell surface of human colon adenocarcinoma cells. AB - Galectin-4, a member of the galectin family, is expressed in the epithelium of the alimentary tract. It has two tandemly repeated carbohydrate recognition domains and specifically binds to an SO3- -->3Galbeta1-->3GalNAc pyranoside with high affinity (Ideo, H., Seko, A., Ohkura, T., Matta, K. L., and Yamashita, K. (2002) Glycobiology 12, 199-208). In this study, we found that galectin-4 binds to glycosphingolipids carrying 3-O-sulfated Gal residues, such as SB1a, SM3, SM4s, SB2, SM2a, and GM1, but not to glycosphingolipids with 3-O-sialylated Gal, such as sLc4Cer, snLc4Cer, GM3, GM2, and GM4, using both an enzyme-linked immunosorbent assay and a surface plasmon resonance assay. A confocal immunocytochemical assay showed that galectin-4 was colocalized with SB1a, GM1, and carcinoembryonic antigen (CEA) in the patches on the cell surface of human colon adenocarcinoma CCK-81 and LS174T cells. This localization was distinct from caveolin/VIP21 localization. Furthermore, immobilized galectin-4 promoted adhesion of CCK-81 cells through the sulfated glycosphingolipid, SB1a. CEA also bound to galectin-4 with KD value of 2 x 10(-8) m by surface plasmon resonance and coimmunoprecipitated with galectin-4 in LS174T cell lysates. These findings suggest that SB1a and CEA in the patches on the cell surface of human colon adenocarcinoma cells could be biologically important ligands for galectin-4. PMID- 15546875 TI - Histone H2A ubiquitination does not preclude histone H1 binding, but it facilitates its association with the nucleosome. AB - Histone H2A ubiquitination is a bulky posttranslational modification that occurs at the vicinity of the binding site for linker histones in the nucleosome. Therefore, we took several experimental approaches to investigate the role of ubiquitinated H2A (uH2A) in the binding of linker histones. Our results showed that uH2A was present in situ in histone H1-containing nucleosomes. Notably in vitro experiments using nucleosomes reconstituted onto 167-bp random sequence and 208-bp (5 S rRNA gene) DNA fragments showed that ubiquitination of H2A did not prevent binding of histone H1 but it rather enhanced the binding of this histone to the nucleosome. We also showed that ubiquitination of H2A did not affect the positioning of the histone octamer in the nucleosome in either the absence or the presence of linker histones. PMID- 15546876 TI - A functional analysis of mouse models of cardiac disease through metabolic profiling. AB - Since the completion of the human and mouse genomes, the focus in mammalian biology has been on assessing gene function. Tools are needed for assessing the phenotypes of the many mouse models that are now being generated, where genes have been "knocked out," "knocked in," or mutated, so that gene expression can be understood in its biological context. Metabolic profiling of cardiac tissue through high resolution NMR spectroscopy in conjunction with multivariate statistics has been used to classify mouse models of cardiac disease. The data sets included metabolic profiles from mouse models of Duchenne muscular dystrophy, two models of cardiac arrhythmia, and one of cardiac hypertrophy. The metabolic profiles demonstrate that the strain background is an important component of the global metabolic phenotype of a mouse, providing insight into how a given gene deletion may result in very different responses in diverse populations. Despite these differences associated with strain, multivariate statistics were capable of separating each mouse model from its control strain, demonstrating that metabolic profiles could be generated for each disease. Thus, this approach is a rapid method of phenotyping mouse models of disease. PMID- 15546877 TI - Effect of amino acid substitutions in the rad50 ATP binding domain on DNA double strand break repair in yeast. AB - The Saccharomyces cerevisiae Rad50-Mre11-Xrs2 complex plays a central role in the cellular response to DNA double strand breaks. Rad50 has a globular ATPase head domain with a long coiled-coil tail. DNA binding by Rad50 is ATP-dependent and the Rad50-Mre11-Xrs2 complex possesses DNA unwinding and endonuclease activities that are regulated by ATP. Here we have examined the role of the Rad50 Walker type A ATP binding motif in DNA double strand break repair by a combination of genetic and biochemical approaches. Replacement of the conserved lysine residue within the Walker A motif with alanine, glutamate, or arginine results in the same DNA damage sensitivity and homologous recombination defect as the rad50 deletion mutation. The Walker A mutations also cause a deficiency in non homologous end-joining. As expected, complexes containing the rad50 Walker A mutant proteins are defective in ATPase, ATP-dependent DNA unwinding, and ATP stimulated endonuclease activities. Although the DNA end-bridging activity of the Rad50-Mre11-Xrs2 complex is ATP-independent, the end-bridging activity of complexes containing the rad50 Walker A mutant proteins is salt-sensitive. These results provide a molecular explanation for the observed in vivo defects of the rad50 Walker mutant strains and reveal a novel ATP-independent function for Rad50 in DNA end-bridging. PMID- 15546878 TI - Live Cell Imaging of ERK and MEK: simple binding equilibrium explains the regulated nucleocytoplasmic distribution of ERK. AB - In response to epidermal growth factor (EGF), the mitogen-activated protein kinase ERK2 translocates into the nucleus. To probe the mechanisms regulating the subcellular localization of ERK2, we used live cell imaging to examine the interaction between MEK1 and ERK2. Fluorescence resonance energy transfer (FRET) studies show that MEK1 and ERK2 directly interact and demonstrate that this interaction in the cytoplasm is largely responsible for cytoplasmic retention of ERK2. Stimulation with EGF caused loss of FRET as ERK separated from MEK and moved into the nucleus. FRET was recovered as ERK returned to the cytosol, indicating ERK reassociation with MEK in the cytoplasm. The EGF-induced transit of ERK through the nucleus was complete within 20 min, and there was no significant movement of MEK into the nucleus. Fluorescence recovery after photobleaching experiments was used to assess the rate of movement of MEK and ERK. The steady-state rate of ERK entry into the nucleus in resting cells was energy-independent and greater than the rate of ERK entry upon EGF stimulation. This suggests that the rate constant for ERK transport across the nuclear membrane is not limiting nuclear entry. Thus, we suggest that the movement of ERK into and out of the nucleus in response to agonist occurs primarily by diffusion and is controlled by interactions with binding partners in the cytosol and nucleus. No evidence of ERK dimerization was detected by FRET methods; the kinetics for nucleocytoplasmic transport were unaffected by mutations in the ERK putative dimerization domain. PMID- 15546880 TI - The centrosomal protein RAS association domain family protein 1A (RASSF1A) binding protein 1 regulates mitotic progression by recruiting RASSF1A to spindle poles. AB - The protein RAS association domain family protein 1A (RASSF1A), which is encoded by a gene that is frequently silenced in many types of sporadic tumor, functions in mitosis as a regulator of the anaphase-promoting complex (APC). With the use of a yeast two-hybrid screen, we identified a human protein, previously designated C19ORF5, that interacts with RASSF1A. This protein, here redesignated RASSF1A-binding protein 1 (RABP1), contains two microtubule-associated protein domains, and its association with RASSF1A was confirmed in mammalian cells by immunoprecipitation and immunofluorescence analyses. RABP1 was found to be localized to the centrosome throughout the cell cycle in a manner dependent on its microtubule-associated protein domains. Ectopic expression of RABP1 induced both stabilization of mitotic cyclins and mitotic arrest at prometaphase in a RASSF1A-dependent manner. It also increased the extent of association between RASSF1A and Cdc20. Conversely depletion of RABP1 by RNA interference prevented both the localization of RASSF1A to the spindle poles as well as its binding to Cdc20, resulting in premature destruction of mitotic cyclins and acceleration of mitotic progression. These findings indicate that RABP1 is required for the recruitment of RASSF1A to the spindle poles and for its inhibition of APC-Cdc20 activity during mitosis. PMID- 15546879 TI - Combination of 5-fluorouracil and N1,N11-diethylnorspermine markedly activates spermidine/spermine N1-acetyltransferase expression, depletes polyamines, and synergistically induces apoptosis in colon carcinoma cells. AB - The thymidylate synthase inhibitor 5-fluorouracil (5-FU) is used widely for chemotherapy of colorectal carcinoma. Recent studies showed that 5-FU affects polyamine metabolism in colon carcinoma cells. We therefore examined whether combinations of 5-FU with drugs that specifically target polyamine metabolism, i.e. N1,N11-diethylnorspermine (DENSPM) or alpha-difluoromethylornithine (DFMO), have synergistic effects in killing HCT116 colon carcinoma cells with wild-type or absent p53. Our results showed that simultaneous 5-FU and DENSPM, a spermine analogue, synergistically increased transcript levels of the polyamine catabolism enzyme spermidine/spermine N1-acetyltransferase, depleted spermine and spermidine, increased acetylated spermidine, and produced synergistic tumor cell apoptosis in both p53 wild-type and p53-null variants. By contrast, simultaneous combination of 5-FU with DFMO, an inhibitor of the polyamine biosynthetic enzyme ornithine decarboxylase, depleted putrescine but did not produce synergistic cell killing. Some pre-treatment and post-treatment regimens of DENSPM and DFMO were antagonistic to 5-FU depending on cellular p53 status. Protein and transcriptome expression analysis showed that combined 5-FU and DENSPM treatment activated caspase 9, but not caspase 3, and significantly suppressed NADH dehydrogenases and cytochrome c oxidases, consistent with the observed increase in hydrogen peroxide, loss of mitochondrial membrane potential, and release of cytochrome c. Our findings demonstrate the importance of the polyamine pathway in 5-FU effects and suggest that the combination of 5-FU with DENSPM has potential for development as therapy for colorectal carcinoma. PMID- 15546881 TI - Selective inhibition of juxtanuclear translocation of protein kinase C betaII by a negative feedback mechanism involving ceramide formed from the salvage pathway. AB - In a previous study, we showed that protein kinase C betaII (PKC betaII) translocated to a novel juxtanuclear compartment as observed in several cell types (Becker, K. P., and Hannun, Y. A. (2003) J. Biol. Chem. 278, 52747-52754). In this study, we noted the absence of this translocation in MCF-7 breast cancer cells, and we examined the mechanisms underlying this selectivity of response. We show that sustained stimulation of PKC betaII with 4beta-phorbol 12-myristate 13 acetate (PMA) resulted in accumulation of ceramide in MCF-7 cells but not in those cells that showed juxtanuclear translocation of PKC betaII. Addition of exogenous ceramides or formation of endogenous ceramide by the action of bacterial sphingomyelinase prevented PMA-induced translocation of PKC betaII in HEK 293 cells. On the other hand, inhibition of ceramide accumulation with fumonisin B1 restored the ability of PMA to induce translocation of PKC betaII in MCF-7 cells. Taken together, the results showed that endogenous ceramide is both necessary and sufficient for preventing juxtanuclear translocation of PKC betaII in response to PMA. Investigation of the mechanisms of ceramide generation in response to PMA revealed that PMA activated the salvage pathway of ceramide formation and not the de novo pathway. This conclusion was based on the following: 1) the ability of fumonisin B1 but not myriocin to inhibit ceramide formation, 2) the ability of PMA to induce increases in palmitate-labeled ceramide only under chase labeling but not acute pulse labeling, 3) the induction of the levels of sphingosine but not dihydrosphingosine in response to PMA, and 4) induction of sphingomyelin hydrolysis in response to PMA. Together, these results define a novel pathway of regulated formation of ceramide, the salvage pathway, and they define a role for this pathway in regulating juxtanuclear translocation of PKC betaII. PMID- 15546882 TI - CC and CX3C chemokines differentially interact with the N terminus of the human cytomegalovirus-encoded US28 receptor. AB - Human cytomegalovirus (HCMV) is the causative agent of life-threatening systemic diseases in immunocompromised patients as well as a risk factor for vascular pathologies, like atherosclerosis, in immunocompetent individuals. HCMV encodes a G-protein-coupled receptor (GPCR), referred to as US28, that displays homology to the human chemokine receptor CCR1 and binds several chemokines of the CC family as well as the CX3C chemokine fractalkine with high affinity. Most importantly, following HCMV infection, US28 activates several intracellular pathways, either constitutively or in a chemokine-dependent manner. In this study, our goal was to understand the molecular interactions between chemokines and the HCMV-encoded US28 receptor. To achieve this goal, a double approach has been used, consisting in the analysis of both receptor and ligand mutants. This approach has led us to identify several amino acids located in the N terminus of US28 that differentially contribute to the high affinity binding of CC versus CX3C chemokines. Additionally, our results highlight the importance of secondary modifications occurring at US28, such as sulfation, for ligand recognition. Finally, the effects of chemokine dimerization and interaction with glycosaminoglycans (GAGs) on chemokine binding and activation of US28 were investigated as well using CCL4 as model ligand. In line with the two-state model describing chemokine/receptor interaction, we show that an aromatic residue in the N-loop region of CCL4 promotes tight binding to US28, whereas receptor activation depends on the presence of the N terminus of CCL4, as shown previously for CCR5. PMID- 15546883 TI - A targeted mutation of Nkd1 impairs mouse spermatogenesis. AB - Nkd1 is an antagonist of the canonical Wnt/beta-catenin signaling pathway. The EF hand motif of Nkd1 is required for its inhibitory function. Early studies suggested that Nkd1 might play important roles in mouse embryonic development and tumorigenesis. We constructed Nkd1(-/-) mice whose Nkd1 protein lacked the EF hand and was unable to inhibit Wnt/beta-catenin signaling. The homozygotes were viable and grew normally, but their fertility in males was reduced. In wild-type adult testes, Nkd1 mRNA was expressed more abundantly in the elongating spermatids than in the round spermatids. Lack of EF-hand caused reductions in the testis weight and sperm count by 30 and 60%, respectively. During testis development, Nkd1 mRNA expression started at the 25th day after birth, coincident with the onset of Wnt1 expression. Nuclear localization of beta-catenin increased in the elongating spermatids, suggesting that the mutant Nkd1 failed to inhibit the Wnt/beta-catenin pathway. These results suggest that deletion of the EF-hand from Nkd1 reduces the number of the elongating spermatids at haploid stage. In contrast, the mutant Nkd1 did not affect intestinal polyposis in Apc(Delta716) mice. PMID- 15546884 TI - DOK1 mediates SHP-2 binding to the alphaVbeta3 integrin and thereby regulates insulin-like growth factor I signaling in cultured vascular smooth muscle cells. AB - Recruitment of the Src homology 2 domain tyrosine phosphatase (SHP-2) to the phosphorylated beta3 subunit of the alphaVbeta3 integrin is required for insulin like growth factor I (IGF-I)-stimulated cell migration and proliferation in vascular smooth muscle cells. Because SHP-2 does not bind directly to beta3, we attempted to identify a linker protein that could mediate SHP-2/beta3 association. DOK1 is a member of insulin receptor substrate protein family that binds beta3 and contains YXXL/I motifs that are potential binding sites for SHP 2. Our results show that IGF-I induces DOK1 binding to beta3 and to SHP-2. Preincubation of cells with synthetic peptides that blocked either DOK1/beta3 or DOK1/SHP-2 association inhibited SHP-2 recruitment to beta3. Expression of a DOK1 mutant that does not bind to beta3 also disrupts SHP-2/beta3 association. As a result of SHP-2/beta3 disruption, IGF-I dependent phosphorylation of Akt and p44/p42 mitogen-activated protein kinase and its ability to stimulate cell migration and proliferation were significantly impaired. These results demonstrate that DOK1 mediates SHP-2/beta3 association in response to IGF-I thereby mediating the effect of integrin ligand occupancy on IGF-IR-linked signaling in smooth muscle cells. PMID- 15546885 TI - Heat shock protein 10 inhibits lipopolysaccharide-induced inflammatory mediator production. AB - Heat shock protein 10 (Hsp10) and heat shock protein 60 (Hsp60) were originally described as essential mitochondrial proteins involved in protein folding. However, both proteins have also been shown to have a number of extracellular immunomodulatory activities. Here we show that purified recombinant human Hsp10 incubated with cells in vitro reduced lipopolysaccharide (LPS)-induced nuclear factor-kappaB activation and secretion of several inflammatory mediators from RAW264.7 cells, murine macrophages, and human peripheral blood mononuclear cells. Induction of tolerance by contaminating LPS was formally excluded as being responsible for Hsp10 activity. Treatment of mice with Hsp10 before endotoxin challenge resulted in the reduction of serum tumor necrosis factor-alpha and RANTES (regulated upon activation, normal T cell expressed and secreted) levels and an elevation of serum interleukin-10 levels. Hsp10 treatment also delayed mortality in a murine graft-versus-host disease model, where gut-derived LPS contributes to pathology. We were unable to confirm previous reports that Hsp10 has tumor growth factor properties and suggest that Hsp10 exerts anti inflammatory activity by inhibiting Toll-like receptor signaling possibly by interacting with extracellular Hsp60. PMID- 15546886 TI - Bass hepcidin synthesis, solution structure, antimicrobial activities and synergism, and in vivo hepatic response to bacterial infections. AB - Bass hepcidin was purified from the gill of hybrid striped bass (Morone chrysops x Morone saxatilis) based on antimicrobial activity against Escherichia coli. This 21-amino acid peptide has 8 cysteines engaged in 4 disulfide bonds and is very similar to human hepcidin, an antimicrobial peptide with iron regulatory properties. To gain insight into potential role(s) of bass hepcidin in innate immunity in fish, we synthesized the peptide, characterized its antimicrobial activities in vitro, determined its solution structure by NMR, and quantified hepatic gene expression in vivo following infection of bass with the fish pathogens, Streptococcus iniae or Aeromonas salmonicida. Its structure is very similar to that of human hepcidin, including the presence of an antiparallel beta sheet, a conserved disulfide-bonding pattern, and a rare vicinal disulfide bond. Synthetic bass hepcidin was active in vitro against Gram-negative pathogens and fungi but showed no activity against key Gram-positive pathogens and a single yeast strain tested. Hepcidin was non-hemolytic at microbicidal concentrations and had lower specific activity than moronecidin, a broad spectrum, amphipathic, alpha-helical, antimicrobial peptide constitutively expressed in bass gill tissue. Good synergism between the bacterial killing activities of hepcidin and moronecidin was observed in vitro. Hepcidin gene expression in bass liver increased significantly within hours of infection with Gram-positive (S. iniae) or Gram-negative (A. salmonicida) pathogens and was 4-5 orders of magnitude above base-line 24-48 h post-infection. Our results suggest that hepcidin plays a key role in the antimicrobial defenses of bass and that its functions are potentially conserved between fish and human. PMID- 15546887 TI - Exogenous antigens are processed through the endoplasmic reticulum-associated degradation (ERAD) in cross-presentation by dendritic cells. AB - Antigen cross-presentation is critical in infectious and tumor immunity where cytotoxic T lymphocytes are induced by dendritic cells specifically equipped with cellular machineries to present exogenous antigens with major histocompatibility complex (MHC) class I molecules. To examine molecular mechanisms of antigen cross presentation, we employed as a model system a murine dendritic cell line DC2.4 capable of presenting soluble antigens such as ovalbumin (OVA) with MHC class I. Here, we demonstrate that exogenously added OVA is accumulated in the endoplasmic reticulum (ER) and late endosomes followed by retrograde transport to the cytoplasm through the Sec61 transporter complexes, and that CHIP functions as an E3 ubiquitin-ligase for OVA degradation by proteasomes. This mechanism is essentially the same as that known as the ER-associated degradation (ERAD) in the quality control of secretary and membrane proteins. PMID- 15546888 TI - N-Acetylcysteine ameliorates lithium-induced renal failure in rats. AB - BACKGROUND: Prolonged lithium treatment may induce progressive deterioration of renal function in humans and experimental animals. N-Acetylcysteine (NAC) has been shown to be effective in the prevention of hypoperfusion and toxin-induced renal failure, but its effect on lithium nephrotoxicity has not been evaluated yet. The purpose of this study was to examine a possible renoprotective effect of NAC against lithium-induced renal failure in a rat model. METHODS: Moderate renal failure was induced in 40 Sprague-Dawley rats using a 5 week protocol including 3 weeks of lithium chloride administration in the drinking water. The animals were divided randomly into two equal groups receiving either 10 mg/kg NAC or saline by two daily intraperitoneal injections. In week 6, the glomerular filtration rate (GFR) was assessed by 99mTechnetium diethylene triaminepentaacetic acid, and serum creatinine, blood urea nitrogen (BUN) and 24 h urinary protein and osmolarity were measured. Kidneys were excised for pathological evaluation. RESULTS: At the end of the lithium protocol, the GFR was significantly higher in the NAC-treated group compared with the control group, 0.92+/-0.35 vs 0.56+/-0.25 ml/min/100 g, respectively, P = 0.002. Serum creatinine and BUN were also significantly lower in the NAC-treated group 1.009+/-0.107 vs 1.143+/-0.118 mg/dl, P = 0.001, and 83.9+/-6.8 vs 88.95+/-7.1 mg/dl, P = 0.28, respectively. The percentages of tubular necrosis and tubular lumen obstruction, evaluated by light microscopy, were significantly lower in the NAC-treated group, P = 0.002 and P = 0.007, respectively. CONCLUSIONS: NAC treatment has a renoprotective effect against lithium-induced renal failure in a rat model. PMID- 15546889 TI - Mesangial cells stimulate differentiation of endothelial cells to form capillary like networks in a three-dimensional culture system. AB - BACKGROUND: There are conflicting results regarding the role of periendothelial mural cells in angiogenesis. In the current study, we investigated the role of mesangial cells (MCs) in endothelial vascularization by using a three-dimensional co-culture system in basement-membrane reconstruct gel (Matrigel). METHODS: Human umbilical vein endothelial cells (ECs) and human MCs were used. In the contact co culture system, ECs and MCs were mixed and then plated together onto Matrigel. In the non-contact co-culture system, MCs were cultured within an intercup chamber, which prevented direct physical contact with the ECs on the Matrigel but allowed both cell types to share culture medium. To visualize ECs and MCs, the cells were labelled with two different fluorescent dyes prior to the co-culture. A capillary like network formation was observed under a fluorescent microscope and confocal microscope, and the length of the network formation was quantified by the image analyzer. RESULTS: ECs barely formed capillary-like networks when cultured alone in growth factor-free medium. However, ECs cultured with MCs in a contact condition remarkably formed capillary-like networks (9.10+/-0.96 vs 0.20+/-0.07 mm/mm2 at 6 h, contact vs ECs alone, P<0.001). Direct contact between ECs and MCs was clearly demonstrated by confocal microscopy. Differentiation into branching capillary-like structures was also observed in the non-contact co-culture system (3.02+/-1.21 mm/mm2 at 6 h, P<0.001 vs ECs alone), but less prominently than in the contact co-culture condition. Vascular endothelial growth factor (VEGF) was secreted from MCs, as determined by enzyme-linked immunosorbent assay and immunofluorescent study. Adding neutralizing antibodies against VEGF into the co culture system partially inhibited capillary network formation. CONCLUSIONS: Our data indicate that MCs help ECs differentiate toward vascularization, in which the direct cell-cell contact between ECs and MCs plays an important role. VEGF is a mediator in this process. PMID- 15546890 TI - Imbalance of growth factor signalling in diabetic kidney disease: is connective tissue growth factor (CTGF, CCN2) the perfect intervention point? PMID- 15546891 TI - Histological and clinical predictors of early and late renal outcome in ANCA associated vasculitis. AB - BACKGROUND: Renal involvement remains a major determinant in antineutrophil cytoplasmic autoantibody-associated small vessel vasculitis (AASV). While some patients may develop persistent renal damage, others have a favourable outcome. METHODS: To identify patients at risk for poor renal outcome, we evaluated 95 renal biopsies (67 initial biopsies and 28 repeat biopsies) of 67 patients with AASV for the presence and extent of active (AI) and chronic (CI) lesions, retrospectively. AI, CI, levels of proteinuria and dose of cyclophosphamide (CYC) were related to renal outcome. RESULTS: Recovery of renal function in patients initially dialysis dependent was associated with a lower CI compared with patients who remained on dialysis (P<0.001), while AI did not differ significantly. In these patients, age <65 years revealed a positive predictive value of 85% for renal function recovery. Patients initially requiring dialysis exhibited a higher AI and CI compared with those who did not. Renal function in long-term follow-up correlated with CI and the amount of proteinuria. This relationship increased with time, exhibiting at 4 years a correlation coefficient of 0.607 for CI (P<0.01) and of 0.775 for proteinuria (P<0.001). Follow-up biopsies showed a more pronounced CI compared with initial biopsies (P<0.001). None of the investigated initial parameters was predictive for renal relapse. However, there was a relationship between dose and duration of CYC and time to relapse. Compared with the initial biopsy, repeat biopsies of eight patients with a creeping serum creatinine in clinical remission showed a decrease of AI (P<0.001) while CI increased rapidly. These patients also had less initial CYC (NS). CONCLUSIONS: These data suggest that in AASV, evaluation of renal histopathology is helpful in predicting early and late renal outcome. Chronicity and proteinuria were the best determinants of poor renal prognosis. Activity may regress under therapy, while chronicity may progress despite treatment. The amount of CYC seems to influence the occurrence of early relapses and renal survival. PMID- 15546892 TI - Duration of end-stage renal disease and kidney transplant outcome. AB - BACKGROUND: Patients nearing end-stage renal disease (ESRD) increasingly choose pre-emptive renal transplant (PRT) to avoid pre-transplant dialysis and to minimize ESRD. Compared with long-term dialysis, PRT has been shown to increase allograft survival. However, the merit of short-term dialysis is not well characterized, and it may be the better medical choice in some patients. The goal of the study was to characterize the relationship between the duration of dialysis vs allograft and patient survival. METHODS: We performed a retrospective nationwide cohort study of all kidney transplants (Tx) between January 1, 1990 and December 31, 1999, with a follow-up period through December 31, 2000. Participants were identified using the United States Renal Data System (USRDS), which tracks all ESRD cases in the nation including patients on dialysis and with kidney Tx. Patients with the history of more than one kidney Tx were excluded. Allograft survival and recipient survival were the primary outcomes of this study. Duration of ESRD as a continuous variable as well as divided into categories (14 days, 15-60 days, 61-180 days, 181-365 days, 1-2 years, 2-3 years, 3-5 years and >5 years) was the primary risk factor of interest. Models were adjusted for multiple donor and recipient factors, including demographics and co morbidities, as well as for Tx procedure characteristics. RESULTS: A total of 81,130 patient records were used for analysis (age 44.1+/-14.3 years, 61% males, 24% black, 29% diabetic, pre-transplant ESRD duration 27.1+/-26.4 months, 26% living donors). ESRD duration, as a continuous variable, is associated with a modest increase in the risk of graft failure over time [hazard ratio (HR) 1.02 per year of ESRD duration, P<0.001]. When ESRD is studied as a categorical variable (duration of 0-14 days vs longer durations), the increased risk of allograft failure reached statistical significance only when the time on dialysis was > or =181 days. The duration of ESRD was a significant risk for recipient death (HR 1.04 per year, P<0.001); however, mortality risk reached statistical significance only when the patient had been on dialysis for > or =1 year. CONCLUSIONS: This study of USRDS records suggests that a short (<6 months) dialysis course has no detrimental effect on graft and patient survival, and should not be deferred if medically indicated. PMID- 15546893 TI - Elevated concentrations of cardiac troponins are associated with severe coronary artery calcification in asymptomatic haemodialysis patients. AB - BACKGROUND: Elevated concentrations of cardiac biomarkers, such as troponins and natriuretic peptides, have been shown to be predictive of poorer long-term cardiovascular outcomes in stable patients with end-stage renal disease (ESRD). However, little is known about the relationship between elevated concentrations of these cardiac markers and underlying coronary artery pathology in these patients. The aim of the present study was to investigate associations between coronary artery calcification (CAC) and the concentrations of cardiac biomarkers in ESRD patients. METHODS: We conducted a cross-sectional study of 38 asymptomatic patients (median age, 54 years; 26 males, 12 females; diabetic, 39%) who were undergoing chronic haemodialysis. In these patients, pre-dialysis circulating concentrations of cardiac troponin T (cTnT), cardiac troponin I (cTnI), creatine kinase-MB (CK-MB) and B-type natriuretic peptide (BNP) were measured. We quantified the level of CAC by multirow spiral computed tomography to obtain a CAC score. CAC scores > or = 400 were defined as being indicative of severe CAC. RESULTS: Severe CAC was detected in 17 patients (45%). The degree of CAC severity was positively associated (P < 0.05) with cTnT concentrations. Thus, 15% of patients had severe CAC in the lowest tertile of cTnT, 50% had severe CAC in the middle third, and 69% in the highest third. Similarly, the degree of severity of CAC was positively associated (P < 0.01) with cTnI concentrations across concentration categories. In contrast, there was no association between the degree of CAC severity and the concentrations of either BNP or CK-MB. A logistic regression analysis revealed that elevated concentrations of cTnT (> or = median vs or = 0.1 ng/ml vs or = 15 yr of age, n = 290) pubertal development. Three SNPs in GNRHR showed modest association with late pubertal development in the trios; among the 506 women, a different SNP was associated with late menarche, and one rare haplotype was associated with early age of menarche. All of the observed associations were relatively modest and only nominally statistically significant; replication is needed to determine their validity. We conclude that genetic variation in GNRH1 and GNRHR is not likely to be a substantial modulator of pubertal timing in the general population. PMID- 15546907 TI - Short-term zoledronic acid treatment increases bone mineral density and marrow clonogenic fibroblast progenitors after allogeneic stem cell transplantation. AB - Although osteoporosis is a relatively common complication after allogeneic stem cell transplantation, the role of bisphosphonates in its management has not yet been completely established. Thirty-two patients who underwent allogeneic stem cell transplantation were prospectively evaluated for bone mineral density (BMD) at the lumbar spine (LS) and femoral neck (FN) after a median period of 12.2 months. Then, 15 of the patients with osteoporosis or rapidly progressing osteopenia (bone loss > 5%/yr) received three monthly doses of 4 mg zoledronic acid iv. Fifteen patients were followed up without treatment, and all 30 patients were reevaluated after 12 months for BMD and bone turnover markers. By using enriched mesenchymal stem cells in the colony-forming units fibroblast (CFU-F) assay, we evaluated the osteogenic stromal lineage. This procedure was performed in both groups of patients at study entry and after 12 months. The average BMD loss was 3.42% at LS and 3.8% at FN during a 1-yr longitudinal evaluation in 32 patients. Subsequently, BMD increased at both LS and FN (9.8 and 6.4%, respectively) in the zoledronic acid-treated cohort. Hydroxyproline excretion decreased, and serum bone-specific alkaline phosphatase increased significantly, whereas serum osteocalcin increase did not reach the limit of significance. A significant increase in CFU-F growth in vitro was induced by in vivo zoledronic acid administration. In the untreated group, no significant change was observed in bone turnover markers, LS BMD (-2.1%), FN BMD (-2.3%), and CFU-F colony number. In conclusion, short-term zoledronic acid treatment consistently improved both LS and FN BMD in transplanted patients who were at high risk for fast and/or persistent bone loss, partly by increasing the osteogenic progenitors in the stromal cell compartment. PMID- 15546908 TI - Toward the development of a test for growth hormone (GH) abuse: a study of extreme physiological ranges of GH-dependent markers in 813 elite athletes in the postcompetition setting. AB - There is a need to develop a test to detect GH abuse by elite athletes. Measured levels of GH in blood or urine, however, provide little information on the GH-IGF I axis. Previous studies have identified a series of indirect markers of GH action that are markedly altered by the administration of GH, but to a lesser degree by acute exercise. This study was undertaken to determine the physiological range of these GH-dependent variables in elite athletes after a competitive event to determine whether such values differ from resting values in normal and athletic subjects and to establish whether any adjustments to this range are required on the basis of age, gender, demographic characteristics, or the nature of the exercise performed. Serum samples were collected from 813 elite athletes (537 males and 276 females; age range, 17-64 yr) from 15 sporting disciplines within 2 h of completion of a major competitive event. IGF-I, IGF binding protein 2 (IGFBP-2), IGFBP-3, acid-labile subunit, and the bone and soft tissue markers, osteocalcin, carboxyl-terminal propeptide of type I procollagen, carboxyl-terminal cross-linked telopeptide of type I collagen, and procollagen type III were measured. Sporting category, gender, age, height, weight, body mass index (BMI), and racial group of the athlete were documented, and results were compared both to normative data and to values obtained from elite athletes under resting conditions. Forty-one percent of IGF-I values in male athletes and 41% of values in female athletes were above the upper limits of 99% reference ranges derived from resting values in a normal population. Postcompetition levels of all variables except carboxyl-terminal propeptide of type I procollagen and carboxyl terminal cross-linked telopeptide of type I collagen differed from resting values. There was a consistent age-dependent fall in measured levels of all variables (P < 0.0001) with the exception of IGFBP-2, which increased with age (P < 0.0001). BMI, but not height, exerted a small, but significant, influence on several variables. After adjustment for age, there were no significant differences in the levels of any of the measured variables between sporting categories. IGFBP-2 and IGFBP-3 were lower in 35 black athletes compared with those in 35 white athletes matched for age, gender, height, BMI, and sporting category. We have demonstrated that there are predictable age-dependent levels of GH-dependent markers in elite athletes that are consistent even at the extremes of physical exertion and that these are independent of sporting category. Normative data applicable to white athletes are provided. This provides important groundwork for the development of a test for GH abuse, although these values may be specific for the reagents and assays used. PMID- 15546909 TI - Limited accuracy of surrogates of insulin resistance during puberty in obese and lean children at risk for altered glucoregulation. AB - This study evaluated the accuracy of surrogate indexes of insulin sensitivity (SI) in children. Surrogates (homeostasis model assessment index of insulin resistance, quick insulin sensitivity index, and 40/insulin ratio index) were cross-sectionally investigated in 66 obese and lean children (17 Tanner stage I, 19 Tanner stage II-III, and 30 Tanner stage IV-V) as indexes of insulin resistance in comparison with the minimal model. The pubertal decrease in SI was found with the minimal model (-47%; P = 0.01), but not with surrogates, which were not correlated to SI. Baseline insulin (Ib) did not mirror the decrease in SI, did not significantly change when plotted against pubertal stage or age, and was not correlated to SI. Ib and surrogates were positively correlated with the body mass index. The disposition index, which quantifies the feedback between SI and insulin release, was widely scattered and decreased during puberty (P = 0.05). The specificity and sensitivity of surrogates as predictors of insulin resistance were poor (e.g. 81.1% and 30.7%, respectively, for the homeostasis model assessment index of insulin resistance). Thus, during puberty, surrogates are not accurate predictors of insulin resistance. Because reference methods are rather expensive and invasive, additional studies of alternative techniques for evaluating SI are needed to allow accurate measurement of insulin resistance in children. PMID- 15546910 TI - Sensitivity to glucocorticoids is decreased in relapsing remitting multiple sclerosis. AB - Endogenous glucocorticoids (GC), which are under control of the hypothalamic pituitary-adrenal axis, play an important role in controlling chronic inflammatory demyelinating diseases, like multiple sclerosis (MS). Increased hypothalamic-pituitary-adrenal axis activity has been found in MS patients and appeared to be negatively associated with acute inflammation. Exogenous GC are frequently used to treat relapses in MS, but the response to this treatment differs among patients, suggesting differences in sensitivity to GC. Previous, relatively small studies investigating GC sensitivity have yielded conflicting results. In the present study, we have investigated GC sensitivity in peripheral blood cells of MS patients (n = 117) and healthy controls (n = 45). GC sensitivity was measured by the in vitro suppressive effect of GC on lipopolysaccharide-stimulated TNF-alpha production. Blood cells of MS patients, especially relapsing remitting MS patients, were less sensitive to GC compared with blood cells of healthy controls. This turned out to be unrelated to previous treatment with exogenous GC expressed as frequency of courses of iv steroids or interval since last course. The use of interferon beta was found to be associated with a lower GC sensitivity. However, after correction for the use of interferon beta, relapsing remitting MS patients remained less sensitive to GC. PMID- 15546911 TI - The impact of dietary protein on calcium absorption and kinetic measures of bone turnover in women. AB - Although high-protein diets induce hypercalciuria in humans, the source of the additional urinary calcium remains unclear. One hypothesis is that the high endogenous acid load of a high-protein diet is partially buffered by bone, leading to increased skeletal resorption and hypercalciuria. We used dual stable calcium isotopes to quantify the effect of a high-protein diet on calcium kinetics in women. The study consisted of 2 wk of a lead-in, well-balanced diet followed by 10 d of an experimental diet containing either moderate (1.0 g/kg) or high (2.1 g/kg) protein. Thirteen healthy women received both levels of protein in random order. Intestinal calcium absorption increased during the high-protein diet in comparison with the moderate (26.2 +/- 1.9% vs. 18.5 +/- 1.6%, P < 0.0001, mean +/- sem) as did urinary calcium (5.23 +/- 0.37 vs. 3.57 +/- 0.35 mmol/d, P < 0.0001, mean +/- sem). The high-protein diet caused a significant reduction in the fraction of urinary calcium of bone origin and a nonsignificant trend toward a reduction in the rate of bone turnover. There were no protein induced effects on net bone balance. These data directly demonstrate that, at least in the short term, high-protein diets are not detrimental to bone. PMID- 15546912 TI - Stem cell antigen-1 is necessary for cell-cycle withdrawal and myoblast differentiation in C2C12 cells. AB - Extracellular signaling pathways regulating myoblast differentiation and cell cycle withdrawal are not completely understood. Stem cell antigen-1 (Sca-1/Ly 6A/E) is a glycosylphosphatidylinositol-anchored membrane protein known for its role in T-cell activation, and recently described as a marker for regeneration competent myoblasts. We previously determined that expression of Sca-1/Ly-6A is transiently upregulated during myocyte cell-cycle withdrawal; however, a specific function for Sca-1 in myogenesis has not been described. Here, we show that Sca-1 expression on the surface of a subpopulation of differentiating C2C12 myoblasts is maximal at the time of cell-cycle withdrawal, and that blocking Sca-1 with monoclonal antibodies or downregulating Sca-1 expression by antisense both promotes proliferation and inhibits myotube formation. Downregulating Sca-1 expression derepresses Fyn at the time of myoblast cell-cycle withdrawal, and dominant-negative and constitutively active Fyn mutants rescue and recapitulate the Sca-1 antisense phenotype, respectively. This suggests a Fyn-mediated mechanism for Sca-1 action. Thus, we demonstrate an unprecedented role for Sca-1 in early myogenesis in C2C12 cells, and propose a novel pathway from the myoblast cell surface to intracellular signaling networks controlling proliferation versus differentiation in mammalian muscle. These findings suggest that, beyond its role as a marker for muscle progenitors, Sca-1 may be an important therapeutic target for promoting muscle regeneration. PMID- 15546914 TI - The lamin CxxM motif promotes nuclear membrane growth. AB - We analyzed the influence of lamins on nuclear envelope growth in cultured Xenopus A6 cells by the overexpression of human lamin A, Xenopus and zebrafish lamins B2 and Drosophila lamins Dm0 and C as GFP fusion proteins. Lamins containing a CxxM motif in their primary sequence (lamins A, B2, Dm0) induced the formation of lobulated nuclei with multi-membrane-layered, highly folded nuclear membranes and intranuclear membrane assemblies, as observed by electron microscopy. Such morphological alterations were not observed with Drosophila lamin C, a lamin without this motif or with a lamin B2 mutant (B2-SxxM) where the cysteine of the CxxM motif is replaced by a serine. Drosophila lamin C mutants containing a CxxM motif behaved like B-type lamins thus confirming that this tetrapeptide is directly involved in the morphological changes we observed. Nuclear membrane proliferation could also be induced by lamin B2 in COS-7 cells and in zebrafish embryos but not by human lamin A in COS-7 cells. We speculate that the human lamin A is incompletely processed in Xenopus A6 cells and therefore behaves in this cell line like a B-type lamin. Our results indicate that the CxxM motif of B-type lamins has a dual function: it mediates lamin targeting to the inner nuclear membrane thereby promoting nuclear membrane growth. PMID- 15546915 TI - Schizosaccharomyces pombe Rgf3p is a specific Rho1 GEF that regulates cell wall beta-glucan biosynthesis through the GTPase Rho1p. AB - Rho1p regulates cell integrity by controlling the actin cytoskeleton and cell wall synthesis. Here, we describe the cloning and characterization of rgf3+, a member of the Rho family of guanine nucleotide exchange factors (Rho GEFs). The rgf3+ gene was cloned by complementation of a mutant (ehs2-1) hypersensitive to drugs that interfere with cell-wall biosynthesis. The rgf3+ gene was found to be essential for cell viability and depletion of Rgf3p afforded phenotypes similar to those obtained following depletion of Rho1p. However, the cell death caused by Rgf3p depletion could be rescued by the presence of 1.2 M sorbitol, whereas depletion of Rho1 was lethal under the same conditions. We show that Rgf3p is a specific Rho1-GEF. The hypersensitivity to drugs affecting the cell wall of the ehs2-1 mutant was suppressed by overexpression of rho1+ but not by any of the other GTPases of the Rho family. Rgf3p interacted with the GDP-bound form of Rho1p and promoted the GDP-GTP exchange. In addition, we show that overexpression of Rgf3p produces multiseptated cells and increases beta-1,3-glucan synthase activity and the amount of cell wall beta-1,3-glucan. Rgf3p localized to the septum and the mRNA level was regulated in a cell-cycle-dependent manner peaking during septation. Our results suggest that Rgf3p acts as a positive activator of Rho1p, probably activating the Rho functions that coordinate cell-wall biosynthesis to maintain cell integrity during septation. PMID- 15546916 TI - LAP2alpha and BAF transiently localize to telomeres and specific regions on chromatin during nuclear assembly. AB - Lamina-associated polypeptide (LAP) 2alpha is a LEM (lamina-associated polypeptide emerin MAN1) family protein associated with nucleoplasmic A-type lamins and chromatin. Using live cell imaging and fluorescence microscopy we demonstrate that LAP2alpha was mostly cytoplasmic in metaphase and associated with telomeres in anaphase. Telomeric LAP2alpha clusters grew in size, formed 'core' structures on chromatin adjacent to the spindle in telophase, and translocated to the nucleoplasm in G1 phase. A subfraction of lamin C and emerin followed LAP2alpha to the core region early on, whereas LAP2beta, lamin B receptor and lamin B initially bound to more peripheral regions of chromatin, before they spread to core structures with different kinetics. Furthermore, the DNA-crosslinking protein barrier-to-autointegration factor (BAF) bound to LAP2alpha in vitro and in mitotic extracts, and subfractions of BAF relocalized to core structures with LAP2alpha. We propose that LAP2alpha and a subfraction of BAF form defined complexes in chromatin core regions and may be involved in chromatin reorganization during early stages of nuclear assembly. PMID- 15546917 TI - Intranuclear membrane structure formations by CaaX-containing nuclear proteins. AB - The nuclear lamina is a protein meshwork lining the nucleoplasmic face of the nuclear envelope. Association of lamins with the inner nuclear membrane is mediated by specific modifications in the CaaX motif at their C-termini. B-type lamins are permanently isoprenylated whereas lamin A loses its modification by a lamin A-specific processing step after incorporation into the lamina. Lamins are differentially expressed during development and tissue differentiation. Here we show that an increased synthesis of lamins B1 and B2 in amphibian oocytes induces the formation of intranuclear membrane structures that form extensive arrays of stacked cisternae. These 'lamin membrane arrays' are attached to the inner nuclear membrane but are not continuous with it. Induction of this membrane proliferation depends on CaaX-specific posttranslational modification. Moreover, in transfected HeLa cells, chimeric GFP containing a nuclear localization signal and a C-terminal CaaX motif of N-Ras induces intranuclear membrane stacks that resemble those induced by lamins and ER-like cisternae that are induced in the cytoplasm upon increased synthesis of integral ER membrane proteins. Implications for the synthesis of CaaX-containing proteins are discussed and the difference from intranuclear fibrous lamina annulate lamellae formations is emphasized. PMID- 15546918 TI - PKA phosphorylation of Src mediates Rap1 activation in NGF and cAMP signaling in PC12 cells. AB - Recent studies suggest that the tyrosine kinase Src plays an important role in the hormonal regulation of extracellular signal-regulated kinases (ERKs) via cyclic AMP (cAMP). Src has also been proposed to mediate signals downstream of nerve growth factor (NGF). Here, we report that the cAMP-dependent protein kinase A (PKA) induced the phosphorylation of Src at residue serine17 (S17) in multiple cell types including PC12, Hek293, AtT-20 and CHO cells. In PC12 cells, Src phosphorylation on S17 participates in the activation of the small G protein Rap1 by both cAMP and NGF. In these cells, Rap1 is required for cAMP/PKA signaling to ERKs and also for the sustained activation of ERKs by NGF. The activation of Rap1 by both cAMP and NGF was blocked by PP2, an inhibitor of Src family kinases, and by a Src mutant incapable of being phosphorylated by PKA (SrcS17A), consistent with the requirement of PKA phosphorylation of Src at S17 in these actions. PP2 and SrcS17A also inhibited the Rap1-dependent activation of ERKs by both agents. These results strongly indicate that PKA phosphorylation of Src at S17 is essential for cAMP and NGF signaling in PC12 cells and identify PKA as an important downstream target of NGF. PKA phosphorylation of Src may therefore be required for Rap1 activation in PC12 cells. PMID- 15546919 TI - ARAP3 is transiently tyrosine phosphorylated in cells attaching to fibronectin and inhibits cell spreading in a RhoGAP-dependent manner. AB - ARAP3 is a GTPase activating protein (GAP) for Rho and Arf GTPases that is implicated in phosphoinositide 3-kinase (PI 3-kinase) signalling pathways controlling lamellipodia formation and actin stress fibre assembly. We have identified ARAP3 as a phosphorylated target of protein tyrosine kinases. In cells, ARAP3 was tyrosine phosphorylated when co-expressed with Src-family kinases (SFKs), upon stimulation with growth factors and during adhesion to the extracellular matrix (ECM) substrate fibronectin. Adhesion-induced phosphorylation of ARAP3 was suppressed by selective inhibitors of Src-family kinases and PI 3-kinase and by a Src dominant interfering mutant. Inducible expression of ARAP3 in HEK293 epithelial cells resulted in increased cell rounding, membrane process formation and cell clustering on ECM substrates. In contrast, ARAP3 dramatically slowed the kinetics of cell spreading on fibronectin but had no effect on cell adhesion. These effects of ARAP3 required a functional Rho GAP domain and were associated with reduced cellular levels of active RhoA and Rac1 but did not require the sterile alpha motif (SAM) or Arf GAP domains. Mutation of two phosphorylation sites, Y1399 and Y1404, enhanced some ARAP3 activities, suggesting that ARAP3 may be negatively regulated by phosphorylation on these tyrosine residues. These results implicate ARAP3 in integrin-mediated tyrosine kinase signalling pathways controlling Rho GTPases and cell spreading. PMID- 15546920 TI - The human histone gene expression regulator HBP/SLBP is required for histone and DNA synthesis, cell cycle progression and cell proliferation in mitotic cells. AB - Histone proteins are essential for chromatin formation, and histone gene expression is coupled to DNA synthesis. In metazoans, the histone RNA binding protein HBP/SLBP is involved in post-transcriptional control of histone gene expression. In vitro assays have demonstrated that human HBP/SLBP is involved in histone mRNA 3' end formation and translation. We have inhibited human HBP/SLBP expression by RNA interference to determine its function during the mitotic cell cycle. Inhibition of HBP/SLBP expression resulted in the inhibition of histone gene expression and DNA synthesis, the inhibition of cell cycle progression in S phase and the inhibition of cell proliferation. These findings indicate that human HBP/SLBP is essential for the coordinate synthesis of DNA and histone proteins and is required for progression through the cell division cycle. PMID- 15546923 TI - Overwintering leaves of a forest-floor fern, Dryopteris crassirhizoma (Dryopteridaceae): a small contribution to the resource storage and photosynthetic carbon gain. AB - BACKGROUND AND AIMS: Dryopteris crassirhizoma is a semi-evergreen fern growing on the floor of deciduous forests. The present study aimed to clarify the photosynthetic and storage functions of overwintering leaves in this species. METHODS: A 2-year experiment with defoliation and shading of overwintering leaves was conducted. Photosynthetic light response was measured in early spring (for overwintering leaves) and summer (for current-year leaves). KEY RESULTS: No nitrogen limitation of growth was detected in plants subjected to defoliation. The number of leaves, their size, reproductive activity (production of sori) and total leaf mass were not affected by the treatment. The defoliation of overwintering leaves significantly reduced the bulk density of rhizomes and the root weight. The carbohydrates consumed by the rhizomes were assumed to be translocated for leaf production. Photosynthetic products of overwintering leaves were estimated to be small. CONCLUSION: Overwintering leaves served very little as nutrient-storage and photosynthetic organs. They partly functioned as a carbon storage organ but by contrast to previous studies, their physiological contribution to growth was found to be modest, probably because this species has a large rhizome system. The small contribution of overwintering leaves during the short-term period of this study may be explained by the significant storage ability of rhizomes in this long-living species. Other ecological functions of overwintering leaves, such as suppression of neighbouring plants in spring, are suggested. PMID- 15546921 TI - Protein kinase B phosphorylation of PIKfyve regulates the trafficking of GLUT4 vesicles. AB - Insulin-stimulated glucose uptake involves the recruitment of the glucose transporter 4 isoform (GLUT4) from an intracellular location to the plasma membrane of fat and muscle cells. Although the activation of the PI3 kinase/protein kinase B (PKB) pathway is central to this effect of insulin, the key substrates for PKB that are involved require identification. Here we report that serine318 on the FYVE domain-containing PtdIns3P 5-kinase (PIKfyve) is a novel substrate for PKB, and show that phosphorylation stimulates the PtdIns3P 5 kinase activity of the enzyme. We also demonstrate that PIKfyve is phosphorylated on serine318 in intact cells in response to insulin, in a PI3-kinase-dependent manner, and that PIKfyve colocalises with a highly motile subpopulation of insulin-regulated aminopeptidase (IRAP)/GLUT4 vesicles. Finally, we demonstrate that overexpression of a PIKfyve[S318A] mutant in 3T3-L1 adipocytes enhances insulin-stimulated IRAP/GLUT4 vesicle translocation to the plasma membrane suggesting a role for PKB-dependent phosphorylation of PIKfyve in insulin regulated IRAP/GLUT4 trafficking. The phosphorylation and activation of PIKfyve by PKB provides a novel signalling paradigm that may link plasma membrane localised PtdIns(3,4,5)P3 signals via a protein kinase cascade to regulated PtdIns(3,5)P2 production, and thereby to the control of trafficking of other membrane cargos. PMID- 15546924 TI - Interactive effects of Al, Ca and other cations on root elongation of rice cultivars under low pH. AB - BACKGROUND AND AIMS: As with other crop species, Al tolerance in rice (Oryza sativa) is widely different among cultivars, and the mechanism for tolerance is unknown. The Ca2+-displacement hypothesis, that is, Al displaces Ca2+ from critical sites in the root apoplast, was predicted to be the essential mechanism for causing Al toxicity in rice cultivars. If displacement of Ca is an essential cause of Al toxicity in rice, Al toxicity may show the same trend as toxicities of elements such as Sr and Ba that are effective in displacing Ca. METHODS: The interactive effects of Al, Ca, Sr and Ba on root elongation of rice cultivars with different Al tolerances were evaluated in hydroponic culture. Al and Ca accumulation in root tips was also investigated. KEY RESULTS AND CONCLUSIONS: Not only Al but also Sr and Ba applications inhibited root growth of rice cultivars under low Ca conditions. As expected, rice cultivars more tolerant of Sr and Ba were also tolerant of Al (japonica > indica). Although Mg application did not affect Sr or Ba toxicity, Mg alleviated Al toxicity to the same level as Ca application. In addition, Ca application decreased the Al content in root tips without displacement. These results suggest that Ca does not have a specific, irreplaceable role in Al toxicity, unlike Sr and Ba toxicities. Alleviation of Al toxicity with increasing concentrations of Ca in rice cultivars is due to increased ionic strength, not due to decreased Al activity. The difference in Al tolerance between indica and japonica cultivars disappears under high ionic strength conditions, suggesting that different electrochemical characteristics of root-tip cells are related to the significant difference in Al tolerance under low ionic strength conditions. PMID- 15546925 TI - Phenotypic correlations among plant parts in Iberian Papilionoideae (Fabaceae). AB - BACKGROUND AND AIMS: Different plant organs may show varying degrees of form diversification or conservatism across phylogenetically related taxa. The present study uses data from a recent systematic study of Iberian Papilionoideae to investigate diversification and covariation in reproductive and vegetative plant parts. The appropriateness of imprecise (but comprehensive) taxonomic quantitative information is tested. METHODS: Organ size covariation and phenotypic correlations were studied among tribes, genera and species. Scale relationships were investigated by Reduced Major Axis regression. Variables used were the maximum dimensions of calyx, corolla, keel petal, fruit, seed, stipule, leaflet and petiole. KEY RESULTS: As regards tribe averages, the length of the corolla and that of calyx correlated positively and significantly. In contrast, pod length was unrelated to corolla size and largely tribe-specific. Within genera, the sizes of calyx, corolla and fruit sometimes covaried linearly (e.g. Lathyrus species) and other times did not (Genista, Astragalus). CONCLUSIONS: Information from taxonomic studies can be useful to establish major phenotypic correlations in plants. Results underscore the implications of tribal ownership in the Papilionoideae and illustrate the extensive morphological diversification of pods relative to flowers in this group. PMID- 15546926 TI - Complex combination of seed dormancy and seedling development determine emergence of Viburnum tinus (Caprifoliaceae). AB - BACKGROUND AND AIMS: The shrub Viburnum tinus is widely distributed in mattoral vegetation of the Mediterranean basin. The purpose of the present study was to classify the seed dormancy type and examine the requirements for embryo growth, root protrusion and shoot emergence. METHODS: Overwintered fruits were collected in western Spain in April 2001 and prepared in three ways: entire pericarp was removed, exocarp and mesocarp were removed or fruits were left intact. Fruits treated in these three ways were subjected to artificial annual temperature cycles or to constant temperature regimes for 1.5 years. KEY RESULTS: Removal of exocarp and mesocarp was necessary for embryo growth and germination. High temperature favoured dormancy alleviation and embryo growth, intermediate to low temperatures favoured root protrusion, and intermediate temperature shoot emergence. There was substantial germination at constant temperature regimes, indicating an overlap between temperature intervals suitable for the different stages of embryo and seedling development. Functionally, V. tinus has the same root and shoot emergence pattern that is described for other Viburnum species considered to have epicotyl dormancy. However, the requirement for high and low temperatures for radicle protrusion and epicotyl emergence, respectively, was missing in V. tinus; these characters are the foundation for the epicotyl dormancy classification. CONCLUSIONS: It is concluded that V. tinus does not have epicotyl dormancy. Instead, there is a combination of a weak morphophysiological dormancy and a slow germination process, where different temperatures during an annual cycle favour different development stages. The present study suggests that the first complete seedlings would emerge in the field 1.5 years after fruit maturation in October, i.e. seed dispersal during winter, embryo growth during the first summer, root protrusion and establishment during the second autumn and winter, and cotyledon emergence during the second spring. PMID- 15546927 TI - Modelling below- and above-ground biomass for non-woody and woody plants. AB - BACKGROUND AND AIMS: Intraspecific relationships between below- and above-ground biomass (MB and MA, respectively) have been studied extensively to evaluate environmental effects on growth and development at the level of the individual plant. However, no current theoretical model for this relationship exists for broad interspecific trends. The aims of this paper are to provide a model and to test its predictions using a recently assembled, large database (1406 data entries for 257 species). METHODS: An allometric model was derived to predict the relationship between MB and MA for non-woody and woody plants based on previously developed scaling relationships for leaf, stem and root standing biomass and annual growth rates. The predictions of this model were tested by comparing the numerical values of predicted scaling exponents (the slopes of log-log regression curves) with those observed for the database. KEY RESULTS AND CONCLUSIONS: For non-woody plants and the juveniles of woody species, the model predicts an isometric scaling relationship (i.e. MB proportional, variant MA). For woody plants, a complex scaling function is predicted. But, for a particular set of biologically reasonable conditions, the model predicts MB proportional, variant MA across woody plants. These predictions accord reasonably well with observed statistical trends when non-woody and woody plants are studied separately (n=1061 and 345 data entries, respectively). Although the reliability of regression formulas to estimate MB based on MA measurements increased with increasing plant size, estimates of MB can be as much as two orders of magnitude off, even when using regression formulas with r2 >>0.90 and F >>53,000. PMID- 15546928 TI - Genetic structure of the endangered plant Neolitsea sericea (Lauraceae) from the Zhoushan archipelago using RAPD markers. AB - BACKGROUND AND AIMS: The Zhoushan archipelago is the largest archipelago in China. It separated from the mainland about 9000 years ago due to rising sea levels and climate change. Because of the long-term influences of human activities, the original forest vegetation on the large islands has been badly damaged and its plant diversity reduced. METHODS: Levels and patterns of genetic diversity in 114 individuals from six natural populations and four cultivated populations of the insular endangered plant Neolitsea sericea (Lauraceae) on the Zhoushan archipelago were assessed using random amplified polymorphic DNA (RAPD) markers. KEY RESULTS: A total of 99 discernible loci were obtained for all populations using ten primers, 50.5 % of which were polymorphic [percentage of polymorphic bands (PPB)=50.5 %]. Despite being a woody, long-lived, perennial, outcrossing and insect-pollinated plant, N. sericea exhibited low levels of genetic variation. The cultivated populations (PPB=18.9 %, HE=0.060, S=0.092) were genetically less diverse than the natural populations (PPB=23.1 %, HE=0.082, S=0.123). Based on analysis of molecular variance, a high degree of among population differentiation was revealed for both natural (0.387) and cultivated populations (0.598). CONCLUSIONS: Removal of plants from the wild for horticulture purposes has eroded the level of genetic variation of N. sericea. Low levels of genetic diversity and a high degree of population differentiation indicate that management strategies should include conservation of natural habitats occupied by all six wild populations, and sampling of germplasm resources from multiple seed sources. PMID- 15546929 TI - Between-year changes in functional gender expression of Palicourea padifolia (Rubiaceae), a distylous, hummingbird-pollinated shrub. AB - BACKGROUND AND AIMS: Because distylous species have two hermaphroditic style length floral morphs, they face two sex allocation problems: the equilibrium morph ratio and the optimal allocation to pollen and seed production in each floral morph. Gender specialization is expected among distylous species when floral morphs differ in reproductive output. However, spatio-temporal variability in female reproductive output between morphs needs to be investigated to fully understand patterns of sexual expression and gender specialization in distylous plants. Between-year variation in flower and fruit production of hummingbird pollinated Palicourea padifolia (Rubiaceae) was examined, focusing on functional gender expression of long- and short-styled morphs and comparing their reproductive performance in five consecutive years (1998-2002). METHODS: Between year variation in inflorescence, floral bud and fruit production was monitored and quantified. These traits were then used as parameters to determine functional gender differences between floral morphs through time. KEY RESULTS: Inflorescence production varied among years but no significant differences were found between floral morphs. Long-styled plants initiated more floral buds per inflorescence every year than short-styled plants, suggesting higher allocation to pollinator attraction and, potentially, an increase in male fitness through pollen donation. Although fruit production was similar between morphs, their functional gender shifted across years. CONCLUSIONS: The gender expression inconsistency across years is surprising because a number of floral characters and attributes that contribute to differently attracting and rewarding effective pollinators in P. padifolia suggest gender specialization. The evidence that morphs of distylous species might specialize in functional gender mostly comes from differences among populations in seed production and non-equilibrium morph ratios based on 1-year field population surveys. The results suggest that more sampling through time is needed to detect gender specialization among distylous species with a perennial habit. PMID- 15546930 TI - Fine-scale clonal structure and diversity within patches of a clone-forming dioecious shrub, Ilex leucoclada (Aquifoliaceae). AB - BACKGROUND AND AIMS: The mode of reproduction (sexual vs. asexual) is likely to have important effects on genetic variation and its spatial distribution within plant populations. An investigation was undertaken of fine-scale clonal structure and diversity within patches of Ilex leucoclada (a clone-forming dioecious shrub). METHODS: Six patches were selected in a 1-ha plot previously established in an old-growth beech forest. Two of the selected patches were composed predominantly of stems with male flowers (male patch), and two contained stems with predominantly female flowers (female patch). The remaining two patches contained stems with male flowers and stems with female flowers in more or less equal proportions (mixed patch). Different genets were distinguished using random amplified polymorphic DNA (RAPD) markers. KEY RESULTS: One hundred and fifty-six genets with different RAPD phenotypes were identified among 1928 stems from the six patches. Among the six patches, the male patches had the lowest clonal diversity, and the mixed patches had the highest. Distribution maps of the genets showed that they extended downhill, reflecting natural layering that occurred when stems were pressed to the ground by heavy snow. In every patch, there were a few large genets with many stems and many small genets with a few stems. CONCLUSION: The differences in clonal diversity among patches may be due to differences in seedling recruitment frequencies. The skewed distribution of genet size (defined as the number of stems per genet) within patches may be due to differences in the timing of germination, or age (with early-establishing genets having clear advantages for acquiring resources) and/or intraspecific competition. PMID- 15546931 TI - Assessment of inequality of root hair density in Arabidopsis thaliana using the Gini coefficient: a close look at the effect of phosphorus and its interaction with ethylene. AB - BACKGROUND AND AIMS: Root hair density (i.e. the number of root hairs per unit root length) in Arabidopsis thaliana varies among individual plants in response to different nutrient stresses. The degree of such variation, defined as inequality, serves as a unique indicator of the uniformity of response within a plant population to nutrient availability. METHODS: Using the Gini coefficient (G) as an inequality index, the inequality of root hair density in Arabidopsis thaliana 'Columbia' was evaluated under conditions of nutrient stresses; in particular the effect of phosphorus and its interaction with ethylene. KEY RESULTS: With decreasing phosphorus concentration, root hair density increased while inequality decreased logarithmically. The addition of the ethylene precursor 1-aminocyclopropane-1-carboxylate (ACC) under high phosphorus increased root hair density and decreased inequality by 7-fold. Inhibition of ethylene action with 1-methylcyclopropene (MCP) and silver thiosulphate (STS) under low phosphorus decreased root hair density, and increased inequality by 9-fold and 4 fold, respectively. The ethylene action inhibitors had little effect on root hair density under high phosphorus, but inequality increased 3-fold in the presence of MCP and decreased 2-fold in the presence of STS. Compared with the control, deficiencies in S, N and K increased inequality of root hair density, whereas deficiencies in P, Ca, B, Mn, Fe, Zn, Cu and Mg decreased inequality. In particular, the inequality of root hair density increased by over 2-fold under deficiencies of N or K, but decreased 14-fold under phosphorus deficiency. CONCLUSIONS: The inequality analysis indicates a strong correlation between prevalent signals from the environment (i.e. phosphorus stress) and the response of the plant, and the role of ethylene in this response. As the environmental signals become stronger, an increasing proportion of individuals respond, resulting in a decrease in variation in responsiveness among individual plants as indicated by reduced inequality. PMID- 15546932 TI - Genetic variability and founder effect in the pitcher plant Sarracenia purpurea (Sarraceniaceae) in populations introduced into Switzerland: from inbreeding to invasion. AB - BACKGROUND AND AIMS: The long-lived and mainly outcrossing species Sarracenia purpurea has been introduced into Switzerland and become invasive. This creates the opportunity to study reactions to founder effect and how a species can circumvent deleterious effects of bottlenecks such as reduced genetic diversity, inbreeding and extinction through mutational meltdown, to emerge as a highly invasive plant. METHODS: A population genetic survey by random amplified polymorphism DNA markers (RAPD) together with historical insights and a field pollination experiment were carried out. KEY RESULTS: At the regional scale, S. purpurea shows low structure (thetast=0.072) due to a recent founder event and important subsequent growth. Nevertheless, multivariate statistical analyses reveal that, because of a bottleneck that shifted allele frequencies, most of the variability is independent among populations. In one population (Tenasses) the species has become invasive and genetic analysis reveals restricted gene flow and family structure (thetast=0.287). Although inbreeding appears to be high (Fis >0.410 from a Bayesian estimation), a field pollination experiment failed to detect significant inbreeding depression upon F1 seed number and seed weight fitness-traits. Furthermore, crosses between unrelated individuals produced F1 seeds with significantly reduced fitness, thus showing local outbreeding depression. CONCLUSIONS: The results suggest that, under restricted gene flow among families, the species may not only have rapidly purged deleterious alleles, but also have undergone some form of selection for inbreeding due to co adaptation between loci. PMID- 15546933 TI - CyDAS: a cytogenetic data analysis system. AB - For statistical analyses in cancer cytogenetics, the genomic changes encoded by the karyotype must be translated into numerical codes. We developed a program, which extracts chromosomal gains and losses as well as breakpoints from the karyotype. The changes are compiled in tables according to the chromosome bands involved and/or depicted in projection to the respective chromosome ideogram. The data are ready to be integrated into further statistical analyses. The program may be run as desktop or Internet application. PMID- 15546934 TI - Application of Petri net theory for modelling and validation of the sucrose breakdown pathway in the potato tuber. AB - MOTIVATION: Because of the complexity of metabolic networks and their regulation, formal modelling is a useful method to improve the understanding of these systems. An essential step in network modelling is to validate the network model. Petri net theory provides algorithms and methods, which can be applied directly to metabolic network modelling and analysis in order to validate the model. The metabolism between sucrose and starch in the potato tuber is of great research interest. Even if the metabolism is one of the best studied in sink organs, it is not yet fully understood. RESULTS: We provide an approach for model validation of metabolic networks using Petri net theory, which we demonstrate for the sucrose breakdown pathway in the potato tuber. We start with hierarchical modelling of the metabolic network as a Petri net and continue with the analysis of qualitative properties of the network. The results characterize the net structure and give insights into the complex net behaviour. PMID- 15546935 TI - Solving and analyzing side-chain positioning problems using linear and integer programming. AB - MOTIVATION: Side-chain positioning is a central component of homology modeling and protein design. In a common formulation of the problem, the backbone is fixed, side-chain conformations come from a rotamer library, and a pairwise energy function is optimized. It is NP-complete to find even a reasonable approximate solution to this problem. We seek to put this hardness result into practical context. RESULTS: We present an integer linear programming (ILP) formulation of side-chain positioning that allows us to tackle large problem sizes. We relax the integrality constraint to give a polynomial-time linear programming (LP) heuristic. We apply LP to position side chains on native and homologous backbones and to choose side chains for protein design. Surprisingly, when positioning side chains on native and homologous backbones, optimal solutions using a simple, biologically relevant energy function can usually be found using LP. On the other hand, the design problem often cannot be solved using LP directly; however, optimal solutions for large instances can still be found using the computationally more expensive ILP procedure. While different energy functions also affect the difficulty of the problem, the LP/ILP approach is able to find optimal solutions. Our analysis is the first large-scale demonstration that LP-based approaches are highly effective in finding optimal (and successive near-optimal) solutions for the side-chain positioning problem. PMID- 15546936 TI - Grid cellware: the first grid-enabled tool for modelling and simulating cellular processes. AB - Modelling and simulation of complex cellular transactions involve development of platforms that understand diverse mathematical representations and are capable of handling large backend computations. Grid Cellware, an integrated modelling and simulation tool, has been developed to precisely address these niche requirements of the modelling community. Grid Cellware implements various pathway simulation algorithms along with adaptive Swarm algorithm for parameter estimation. For enchanced computational productivity Grid Cellware uses grid technology with Globus as the middleware. PMID- 15546937 TI - Multiple sequence alignment with user-defined constraints at GOBICS. AB - Most multi-alignment methods are fully automated, i.e. they are based on a fixed set of mathematical rules. For various reasons, such methods may fail to produce biologically meaningful alignments. Herein, we describe a semi-automatic approach to multiple sequence alignment where biological expert knowledge can be used to influence the alignment procedure. The user can specify parts of the sequences that are biologically related to each other; our software program uses these sites as anchor points and creates a multiple alignment respecting these user defined constraints. By using known functionally, structurally or evolutionarily related positions of the input sequences as anchor points, our method can produce alignments that reflect the true biological relationships among the input sequences more accurately than fully automated procedures can do. PMID- 15546938 TI - PermutMatrix: a graphical environment to arrange gene expression profiles in optimal linear order. AB - PermutMatrix is a work space designed to graphically explore gene expression data. It relies on the graphical approach introduced by Eisen and also offers several methods for the optimal reorganization of rows and columns of a numerical dataset. For example, several methods are proposed for optimal reorganization of the leaves of a hierarchical clustering tree, along with several seriation or unidimensional scaling methods that do not require any preliminary hierarchical clustering. This program, developed for MS Windows, with MS-Visual C++, has a clear and efficient graphical interface. Large datasets can be thoroughly and quickly analyzed. PMID- 15546939 TI - Probabilities of spurious connections in gene networks: application to expression time series. AB - MOTIVATION: The reconstruction of gene networks from gene-expression microarrays is gaining popularity as methods improve and as more data become available. The reliability of such networks could be judged by the probability that a connection between genes is spurious, resulting from chance fluctuations rather than from a true biological relationship. RESULTS: Unlike the false discovery rate and positive false discovery rate, the decisive false discovery rate (dFDR) is exactly equal to a conditional probability without assuming independence or the randomness of hypothesis truth values. This property is useful not only in the common application to the detection of differential gene expression, but also in determining the probability of a spurious connection in a reconstructed gene network. Estimators of the dFDR can estimate each of three probabilities: (1) The probability that two genes that appear to be associated with each other lack such association. (2) The probability that a time ordering observed for two associated genes is misleading. (3) The probability that a time ordering observed for two genes is misleading, either because they are not associated or because they are associated without a lag in time. The first probability applies to both static and dynamic gene networks, and the other two only apply to dynamic gene networks. PMID- 15546940 TI - SlidingBayes: exploring recombination using a sliding window approach based on Bayesian phylogenetic inference. AB - We developed a software tool (SlidingBayes) for recombination analysis based on Bayesian phylogenetic inference. Sliding-Bayes provides a powerful approach for detecting potential recombination, especially between highly divergent sequences and complex HIV-1 recombinants for which simpler methods like neighbor joining (NJ) may be less powerful. SlidingBayes guides Markov Chain Monte Carlo (MCMC) sampling performed by MrBayes in a sliding window across the alignment (Bayesian scanning). The tool can be used for nucleotide and amino acid sequences and combines all the modeling possibilities of MrBayes with the ability to plot the posterior probability support for clustering of various combinations of taxa. PMID- 15546941 TI - BRAGI: linking and visualization of database information in a 3D viewer and modeling tool. AB - BRAGI is a well-established package for viewing and modeling of three-dimensional (3D) structures of biological macromolecules. A new version of BRAGI has been developed that is supported on Windows, Linux and SGI. The user interface has been rewritten to give the standard 'look and feel' of the chosen operating system and to provide a more intuitive, easier usage. A large number of new features have been added. Information from public databases such as SWISS-PROT, InterPro, DALI and OMIM can be displayed in the 3D viewer. Structures can be searched for homologous sequences using the NCBI BLAST server. PMID- 15546942 TI - A rare case of streptomycin-induced toxic epidermal necrolysis in a patient with tuberculosis: a therapeutic dilemma. AB - OBJECTIVE: To report a case of streptomycin-induced toxic epidermal necrolysis (TEN). CASE SUMMARY: A 55-year-old woman was admitted for treatment of active pulmonary tuberculosis (TB). She was given standard oral anti-TB chemotherapy including isoniazid, rifampin, pyrazinamide, and streptomycin. On the fourth day of therapy, she experienced high fever at 39 degrees C, chills, vomiting, pruritus, and diffuse erythema, followed by extensive bullae formation and skin denudation. Diagnosis of TEN was considered, and all anti-TB drugs were discontinued. Skin biopsy disclosed complete epidermal necrosis with dermal epidermal cleavage and absence of inflammatory infiltrate, highly suggestive of TEN. The patient was transferred to the intensive care unit. Her general condition and skin lesions improved. A staged-fashion exposure test to the 4 anti TB drugs allowed the incrimination of streptomycin as the offending agent. DISCUSSION: Anti-TB drugs, mainly rifampin, ethambutol, and isoniazid, have been incriminated in TEN. Streptomycin-induced TEN remains an extremely rare event. However, minor allergic skin reactions (rash, urticaria) have been described with this drug. Our patient presents a rare case of streptomycin-related TEN. Even though dangerous, a step-wise exposure test was necessary to allow safe treatment of active pulmonary TB. It also provided a strong argument of a cause-effect relationship between TEN and streptomycin. An objective causality assessment using the Naranjo rating scale revealed that the adverse drug event was highly probable. CONCLUSIONS: Streptomycin should be added to the list of drugs that induce TEN. PMID- 15546943 TI - Coadministration of gatifloxacin and multivitamin preparation containing minerals: potential treatment failure in an elderly patient. AB - OBJECTIVE: To report a case of probable treatment failure in a patient receiving gatifloxacin and a multivitamin preparation containing minerals. CASE SUMMARY: A 77-year-old white woman was prescribed gatifloxacin for hospital-acquired bacterial pneumonia. She was also receiving calcium carbonate 500 mg twice daily and a multivitamin preparation containing minerals once a day. Three days after gatifloxacin was started, the patient was still febrile, coughing, and not responding clinically. It was noted that nurses were administering the gatifloxacin tablet at the same time as the multivitamin tablet. The time of administration for gatifloxacin was changed to 6 hours after the patient received her multivitamin preparation. Two days later, she clinically improved. DISCUSSION: Studies have shown that the bioavailability of gatifloxacin is decreased with concurrent administration of antacids containing aluminum or magnesium; dietary supplements containing zinc, magnesium, and iron; multivitamin preparations containing minerals; and sucralfate. An objective causality assessment revealed that the adverse drug effect was probable. CONCLUSIONS: This case illustrates the need to recognize this potential interaction and to know how to avoid possible treatment failure. PMID- 15546944 TI - Adverse reactions during imatinib and lansoprazole treatment in gastrointestinal stromal tumors. AB - OBJECTIVE: To report the case of a patient affected by gastrointestinal stromal tumors (GIST) who developed cutaneous adverse drug reactions during treatment with imatinib and lansoprazole. CASE SUMMARY: After 2 months of treatment with imatinib 400 mg/day, a 60-year-old white female affected by GIST developed bilateral palpebral edema with hyperemic conjunctivae and labial edema when lansoprazole 15 mg/day was introduced to treat dyspeptic symptomatology. Treatment was discontinued, and on reintroduction of both drugs, the patient developed Stevens-Johnson syndrome. Two months later, generalized cutaneous reactions appeared immediately following reintroduction of low-dose imatinib with corticosteroid plus lansoprazole treatment. After discontinuation of all drugs, with the exception of the corticosteroid, the progression of cutaneous lesions stopped. DISCUSSION: The use of imatinib is commonly associated with a high dose dependent rate of rash and edema. Several cases of Stevens-Johnson syndrome have also been described, although not in patients affected by GIST. Severe skin reactions have been reported for lansoprazole including erythema multiforme, Stevens-Johnson syndrome, and toxic epidermal necrolysis. Applying Naranjo's algorithm, the adverse events were considered possible due to imatinib and probable due to lansoprazole. CONCLUSIONS: On the basis of the data reported, we conclude that the adverse reactions described may be attributed to either drug alone. However, combined use of drugs may increase the risk of onset of these adverse reactions due to a potential drug interaction involving CYP3A4. PMID- 15546945 TI - Assessment for chronic kidney disease service in high-risk patients at community health clinics. AB - BACKGROUND: Chronic kidney disease (CKD) poses significant public health concerns. Early identification and interventions can help prevent or slow progression to end-stage renal disease. OBJECTIVE: To characterize CKD in high risk indigent patients in a primary care setting and evaluate opportunities for pharmacists to work collaboratively with physicians to improve medication use and CKD patient outcomes. METHODS: Medical records of 200 patients with diabetes mellitus and/or hypertension were reviewed by the clinical pharmacist. Estimated glomerular filtration rate (creatinine clearance [Cl(cr)]) and urinalysis were used to identify and stage CKD according to published guidelines. Glycosylated hemoglobin concentrations and blood pressures were recorded. The pharmacist evaluated medications for possible drug-related problems (DRPs), made therapeutic recommendations, and evaluated the acceptance rate by physicians. RESULTS: One hundred nineteen patients met inclusion criteria, and a total of 68.9% met CKD criteria: stage 1, 16.0%; stage 2, 20.2%; stage 3, 25.2%; stage 4, 1.7%; stage 5, 0.8%; and not stageable, 5.0%. A total of 381 DRPs were identified, averaging 3.2 (1.7) per patient (range 0-11). The number of DRPs correlated with Cl(cr) (r = 0.25; p = 0.007). Therapeutic recommendations included change of drug, dose and/or interval adjustment of the current drug, discontinuation of nonsteroidal antiinflammatory drugs, additional laboratory monitoring, meeting goal blood pressure and glycosylated hemoglobin, adding renoprotective drug and/or low-dose aspirin, and nephrologist referral. Fewer than half (40.9%) of the recommendations were accepted or accepted with modifications, and an approximately equal percentage were not accepted by the physicians. CONCLUSIONS: CKD prevalence was high among the patients evaluated here. New guidelines are available to assist in managing CKD ambulatory patients. Pharmacist collaboration with physicians may optimize CKD screening in high-risk patients and improve medication usage. PMID- 15546946 TI - Television and adolescent use of over-the-counter analgesic agents. AB - BACKGROUND: Portrayals of the use of over-the-counter (OTC) analgesics on television may stimulate their use. OBJECTIVE: To explore the association between taking OTC analgesics and television viewing by adolescents. METHODS: A standardized self-administered questionnaire among first-year (mean age 13.16 y) and fourth-year (mean age 16.37 y) secondary students in Flanders, Belgium (n = 2546) was administered in a school setting. The independent variable was television viewing; control variables were school year, gender, drinking alcohol, days off sick, computer gaming, and Internet use. The main outcome variable was monthly use of OTC analgesics. RESULTS: On average, respondents watched 3 hours 18 minutes of television per day (boys more than girls, first-year students more than fourth-year students). Per additional hour of television per day, the odds that respondents were regular users of analgesics were 1.16 times higher (95% CI 1.08 to 1.24). For the 10% heaviest viewers, the odds were 2.30 times higher (95% CI 1.25 to 4.24) compared with the 10% lightest viewers. Odds of OTC analgesic use were higher for students reporting more sick days in the past year (OR 1.38, 95% CI 1.28 to 1.49), regular users of alcohol (OR 1.33, 95% CI 1.17 to 1.50), and girls (OR 1.92, 95% CI 1.35 to 2.73). CONCLUSIONS: A relationship was found between watching television and the use of OTC analgesics, even after controlling for gender and lifestyle measures. More research is needed to establish whether this relationship is causal. PMID- 15546947 TI - The single nucleotide polymorphism Ser128Arg in the E-selectin gene is associated with enhanced coagulation during human endotoxemia. AB - The single nucleotide polymorphism (SNP) Ser128Arg in the E-selectin gene is overrepresented in certain patient groups with atherosclerosis or restenosis. We hypothesized and tested whether it may affect cytokine-induced levels of soluble (s) E-selectin, or be associated with proinflammatory or procoagulant properties in a well-standardized inflammation model. Healthy male volunteers (n = 157) received a lipopolysaccharide (LPS) infusion and were genotyped for the S128R SNP, and outcome parameters were measured by enzyme immunoassays and real-time polymerase chain reaction (RT-PCR, Taqman). The S128R SNP had no pronounced effects on basal or inducible sE-selectin levels, or levels of tumor necrosis factor or interleukin-6. However, carriers of the S128R SNP had 20% higher monocyte counts at 24 hours after LPS infusion. Importantly, the S128R allele enhanced thrombin generation by 50% to 80%, as measured by prothrombin fragment F(1+2) (P < .01), and hence fibrin formation (D-dimer) 2-fold (P = .01 to P = .002). However, tissue factor (TF) mRNA levels were not affected. The S128R E selectin genotype is associated with procoagulant effects in a human model of endotoxin-induced, TF-triggered coagulation. This could contribute to its linkage with various thrombotic cardiovascular disorders. PMID- 15546948 TI - Tumor antigen processing and presentation depend critically on dendritic cell type and the mode of antigen delivery. AB - Dendritic cells (DCs) are being evaluated for cancer immunotherapy due to their unique ability to induce tumor-directed T-cell responses. Here we report that the type of human DC, the mode of activation, and the strategy for delivery of antigen are 3 critical factors for efficient stimulation of tumor-specific CD8+ and CD4+ T cells. Only CD1c+ blood DCs and monocyte-derived DCs (MoDCs) were capable of presenting epitopes of the full-length tumor antigen NY-ESO-1 on both major histocompatibility complex (MHC) class I (cross-presentation) and MHC II, whereas plasmacytoid DCs were limited to MHC II presentation. Cross-presentation was inefficient for soluble protein, but highly efficient for antigen-antibody immune complexes (NY-ESO-1/IC) and for protein formulated with ISCOMATRIX adjuvant (NY-ESO-1/IMX). DC activation with CD40L further enhanced cross presentation efficiency. The mode of antigen delivery was found to be a determining factor for cytosolic proteolysis by DCs. Immune complexes (ICs) targeted a slow, proteasome-dependent cross-presentation pathway, whereas ISCOMATRIX (IMX) targeted a fast, proteasome-independent pathway. Both cross presentation pathways resulted in a long-lived, T-cell stimulatory capacity, which was maintained for several days longer than for DCs pulsed with peptide. This may provide DCs with ample opportunities for sensitizing tumor-specific T cells against a broad array of tumor antigen epitopes in lymph nodes. PMID- 15546949 TI - Differential requirements for calcium and Src family kinases in platelet GPIIb/IIIa activation and thromboxane generation downstream of different G protein pathways. AB - G(12/13) or G(q) signaling pathways activate platelet GPIIb/IIIa when combined with G(i) signaling. We tested whether combined G(i) and G(z) pathways also cause GPIIb/IIIa activation and compared the signaling requirements of these events. Platelet aggregation occurred by combined stimulation of G(i) and G(z) pathways in human platelets and in P2Y1-deficient and G alpha(q)-deficient mouse platelets, confirming that the combination of G(i) and G(z) signaling causes platelet aggregation. When G(i) stimulation was combined with G(z) stimulation, there was a small mobilization of intracellular calcium. Chelation of intracellular calcium decreased the extent of this platelet aggregation, whereas it abolished the G(q) plus G(i)-mediated platelet aggregation. Costimulation of G(i) plus G(z) pathways also caused thromboxane generation that was dependent on outside-in signaling and was inhibited by PP2, a Src family tyrosine kinase inhibitor. Src family tyrosine kinase inhibitors also inhibited platelet aggregation and decreased the PAC-1 binding caused by costimulation of G(i) and G(z) signaling pathways in aspirin-treated platelets. However, Src family kinase inhibitors did not affect G(q) plus G(i)-mediated platelet aggregation. We conclude that the combination of G(i) plus G(z) pathways have different requirements than G(q) plus G(i) pathways for calcium and Src family kinases in GPIIb/IIIa activation and thromboxane production. PMID- 15546950 TI - Only the CD62L+ subpopulation of CD4+CD25+ regulatory T cells protects from lethal acute GVHD. AB - CD4+CD25+ regulatory T (Treg) cells are potent modulators of alloimmune responses. In murine models of allogeneic bone marrow transplantation, adoptive transfer of donor CD4+CD25+ Treg cells protects recipient mice from lethal acute graft-versus-host disease (aGVHD) induced by donor CD4+CD25- T cells. Here we examined the differential effect of CD62L+ and CD62L- subsets of CD4+CD25+ Treg cells on aGVHD-related mortality. Both subpopulations showed the characteristic features of CD4+CD25+ Treg cells in vitro and did not induce aGVHD in vivo. However, in cotransfer with donor CD4+CD25- T cells, only the CD62L+ subset of CD4+CD25+ Treg cells prevented severe tissue damage to the colon and protected recipients from lethal aGVHD. Early after transplantation, a higher number of donor-type Treg cells accumulated in host mesenteric lymph node (LN) and spleen when CD4+CD25+CD62L+ Treg cells were transferred compared with the CD62L- subset. Subsequently, CD4+CD25+CD62L+ Treg cells showed a significantly higher capacity than their CD62L- counterpart to inhibit the expansion of donor CD4+CD25- T cells. The ability of Treg cells to efficiently enter the priming sites of pathogenic allo-reactive T cells appears to be a prerequisite for their protective function in aGVHD. PMID- 15546951 TI - Prognostic value of pretransplantation host thymic function in HLA-identical sibling hematopoietic stem cell transplantation. AB - Thymic function is critical for immune reconstitution after hematopoietic stem cell transplantation (HSCT). We evaluated recipient thymic function before HSCT by quantifying T-cell receptor excision circles (TRECs) in pretransplantation peripheral blood lymphocytes from 102 patients who received HSCs from an HLA identical sibling for malignant (n = 87) or nonmalignant diseases (n = 15). Median TREC value before transplantation was 257 TRECs per 150,000 CD3+ cells (range, 0-42,746). We assessed 172 TRECs per 150,000 CD3+ cells as the most discriminating TREC value for survival in a first cohort of patients (n = 62). This cut-off was validated in a second independent prospective group of 40 patients. In the 102 patients, a TREC value greater than or equal to 172 was associated with a better survival (P < .000 01), a decreased incidence of grade II-IV acute graft-versus-host disease (GVHD; P = .017), chronic GVHD (P = .023), and bacterial (P = .003) and cytomegalovirus (CMV) infection (P = .024). In a multivariate analysis, low pretransplantation TREC values were associated with a higher incidence of CMV infection (hazard ratio [HR] = 2.0, P = .06) and severe bacterial infections (HR = 2.8, P = .036). Finally, high TREC values (HR = 6.6, P = .002) and ABO compatibility (HR = 2.7, P = .02) were associated with a better survival. Therefore, recipient host thymic function assessment could be helpful in predicting HSCT outcome and identifying patients who require a close immunologic monitoring. PMID- 15546952 TI - Increased expression of the Abcg2 transporter during erythroid maturation plays a role in decreasing cellular protoporphyrin IX levels. AB - ABCG2/BCRP is a member of the adenosine triphosphate-binding cassette (ABC) transporter family and is expressed in intestine, kidney, and liver, where it modulates the absorption and excretion of xenobiotic compounds. ABCG2 is also expressed in hematopoietic stem cells and erythroid cells; however, little is known regarding its role in hematopoiesis. Abcg2 null mice have increased levels of protoporphyrin IX (PPIX) in erythroid cells, yet the mechanism for this remains uncertain. We have found that Abcg2 mRNA expression was up-regulated in differentiating erythroid cells, coinciding with increased expression of other erythroid-specific genes. This expression pattern was associated with significant amounts of ABCG2 protein on the membrane of mature peripheral blood erythrocytes. Erythroid cells engineered to express ABCG2 had significantly lower intracellular levels of PPIX, suggesting the modulation of PPIX level by ABCG2. This modulating activity was abrogated by treatment with a specific ABCG2 inhibitor, Ko143, implying that PPIX may be a direct substrate for the transporter. Taken together, our results demonstrate that ABCG2 plays a role in regulating PPIX levels during erythroid differentiation and suggest a potential role for ABCG2 as a genetic determinant in erythropoietic protoporphyria. PMID- 15546953 TI - The nitric oxide pathway modulates hemangioblast activity of adult hematopoietic stem cells. AB - We have previously established a model inducing hematopoietic stem cell (HSC) production of circulating endothelial progenitor cells (EPCs) to revascularize ischemic injury in adult mouse retina. The unique vascular environment of the retina results in new blood vessel formation primarily from HSC-derived EPCs. Using mice deficient (-/-) in inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS), we show that vessel phenotype resulting from hemangioblast activity can be altered by modulation of the NO/NOS pathway. iNOS-/- or eNOS-/- animals were engrafted with wild-type (WT) HSCs expressing green fluorescence protein (gfp+) and subjected to our adult retinal ischemia model. WT hemangioblast activity in adult iNOS-/- recipients resulted in the formation of highly branched blood vessels of donor origin, which were readily perfused indicating functionality. In contrast, eNOS-/- recipients produced relatively unbranched blood vessels with significant donor contribution that were difficult to perfuse, indicating poor functionality. Furthermore, eNOS-/- chimeras had extensive gfp+ HSC contribution throughout their vasculature without additional injury. This neovascularization, via EPCs derived from the transplanted HSCs, reveals that the NO pathway can modulate EPC activity and plays a critical role in both blood vessel formation in response to injury and normal endothelial cell maintenance. PMID- 15546954 TI - Aspergillus fumigatus suppresses the human cellular immune response via gliotoxin mediated apoptosis of monocytes. AB - Aspergillus fumigatus (AF) is a ubiquitous mold and is the most common cause of invasive aspergillosis, an important source of morbidity and mortality in immunocompromised hosts. Using cytokine flow cytometry, we assessed the magnitude of functional CD4+ and CD8+ T-cell responses following stimulation with Aspergillus antigens. Relative to those seen with cytomegalovirus (CMV) or superantigen stimulation, responses to Aspergillus antigens were near background levels. Subsequently, we confirmed that gliotoxin, the most abundant mycotoxin produced by AF, was able to suppress functional T-cell responses following CMV or staphylococcal enterotoxin B (SEB) stimulation. Additional studies demonstrated that crude AF filtrates and purified gliotoxin inhibited antigen-presenting cell function and induced the preferential death of monocytes, leading to a marked decrease in the monocyte-lymphocyte ratio. Analysis of caspase-3 activation confirmed that gliotoxin preferentially induced apoptosis of monocytes; similar effects were observed in CD83+ monocyte-derived dendritic cells. Importantly, the physiologic effects of gliotoxin in vitro were observed below concentrations recently observed in the serum of patients with invasive aspergillosis. These studies suggest that the production of gliotoxin by AF may constitute an important immunoevasive mechanism that is mediated by direct effects on antigen presenting cells and both direct and indirect effects on T cells. PMID- 15546955 TI - Donor T-cell production of RANTES significantly contributes to the development of idiopathic pneumonia syndrome after allogeneic stem cell transplantation. AB - Idiopathic pneumonia syndrome (IPS) is a major cause of mortality following allogeneic stem cell transplantation (allo-SCT). Clinical and experimental data support a role for conditioning-induced inflammation and alloreactive T-cell responses in IPS pathophysiology, but the mechanisms by which donor leukocytes are ultimately recruited to the lung are not fully understood. RANTES is a chemokine ligand that is up-regulated during inflammation and promotes the recruitment of T cells and macrophages to sites of tissue damage. Using a lethally irradiated murine SCT model (B6 --> B6D2F1), we evaluated the role of donor leukocyte-derived RANTES in the development of IPS. Pulmonary mRNA and protein levels of RANTES were significantly elevated in allo-SCT recipients compared to syngeneic controls and were associated with enhanced mRNA expression of CCR5 and CCR1 and with inflammatory cell infiltration into the lung. Allo-SCT with RANTES-/- donor cells significantly decreased IPS and improved survival. Combinations of allogeneic wild-type or RANTES-/- bone marrow with wild-type or RANTES-/- T cells demonstrated that the expression of RANTES by donor T cells was critical to the development of lung injury after SCT. These data reveal that donor T cells can help regulate leukocyte recruitment to the lung after allo-SCT and provide a possible mechanism through which inflammation engendered by SCT conditioning regimens is linked to allo-specific T-cell responses during the development of IPS. PMID- 15546956 TI - Autologous HSCT for severe progressive multiple sclerosis in a multicenter trial: impact on disease activity and quality of life. AB - Hematopoietic stem cell transplantation (HSCT) has been proposed for the treatment of severe multiple sclerosis (MS). In a phase 2 multicenter study we selected 19 non-primary progressive MS patients showing high disease activity on the basis of both brain magnetic resonance imaging (MRI) and sustained clinical deterioration despite conventional treatments. After stem cell mobilization with cyclophosphamide (CY) and filgrastim, patients were conditioned with BCNU (1,3 bis(2-chloroethyl)-1-nitrosourea), cytosine arabinoside, etoposide, and melphalan (BEAM) followed by antithymocyte globulin (ATG). Unmanipulated peripheral blood stem cells (PBSCs) were then infused. No maintenance treatment was administered with a median follow-up of 36 months (range, 12 to 72 months). All patients showed clinical stabilization or improvement; 3 subsequently deteriorated, 1 beyond the baseline. No MRI active lesions were detected after the HSCT except in 1 patient who showed a new lesion at 4.5 years. Infections were limited and restricted to 3 months after HSCT. Health-related quality of life was assessed through the 54-item MS quality of life (MSQOL-54) questionnaire, showing a statistically significant improvement in both composite scores and in most of the individual domains. HSCT is able to induce a prolonged clinical stabilization in severe progressive MS patients, resulting in both sustained treatment-free periods and quality of life improvement. PMID- 15546957 TI - Arginase I is constitutively expressed in human granulocytes and participates in fungicidal activity. AB - The balance of arginine metabolism via nitric oxide synthase (NOS) or arginase is an important determinant of the inflammatory response of murine macrophages and dendritic cells. Here we analyzed the expression of the isoform arginase I in human myeloid cells. Using healthy donors and patients with arginase I deficiency, we found that in human leukocytes arginase I is constitutively expressed only in granulocytes and is not modulated by a variety of proinflammatory and anti-inflammatory stimuli in vitro. We demonstrate that arginase I is localized in azurophil granules of neutrophils and constitutes a novel antimicrobial effector pathway, likely through arginine depletion in the phagolysosome. Our findings demonstrate important differences between murine and human leukocytes with respect to regulation and function of arginine metabolism via arginase. PMID- 15546958 TI - A rich chemokine environment strongly enhances leukocyte migration and activities. AB - The migration of leukocytes in immune surveillance and inflammation is largely determined by their response to chemokines. While the chemokine specificities and expression patterns of chemokine receptors are well defined, it is still a matter of debate how leukocytes integrate the messages provided by different chemokines that are concomitantly produced in physiologic or pathologic situations in vivo. We present evidence for a novel regulatory mechanism of leukocyte trafficking. Our data are consistent with a mode of action where CC-chemokine receptor 7 (CCR7) agonists and unrelated, nonagonist chemokines first form a heteromeric complex, in the presence of which the triggering of CCR7 can occur at a much lower agonist concentration. The increase is synergistic and can be evoked by many but not all chemokines. Chemokine-induced synergism might provide an amplification system in "chemokine-rich" tissues, rendering leukocytes more competent to respond to migratory cues. PMID- 15546959 TI - Rapamycin attenuates vascular wall inflammation and progenitor cell promoters after angioplasty. AB - Rapamycin combines antiproliferative and antiinflammatory properties and reduces neointima formation after angioplasty in patients. Its effect on transcriptional programs governing neointima formation has not yet been investigated. Here, we systematically analyzed the effect of rapamycin on gene expression during neointima formation in a human organ culture model. After angioplasty, renal artery segments were cultured for 21 or 56 days in absence or presence of 100 ng/ml rapamycin. Gene expression analysis of 2312 genes revealed 264 regulated genes with a peak alteration after 21 days. Many of those were associated with recruitment of blood cells and inflammatory reactions of the vessel wall. Likewise, chemokines and cytokines such as M-CSF, IL-1beta, IL-8, beta thromboglobulin, and EMAP-II were found up-regulated in response to vessel injury. Markers indicative for a facilitated recruitment and stimulation of hematopoetic progenitor cells (HPC), including BST-1 and SDF-1, were also induced. In this setting, rapamycin suppressed the coordinated proadhesive and proinflammatory gene expression pattern next to down-regulation of genes related to metabolism, proliferation, and apoptosis. Our study shows that mechanical injury leads to induction of a proinflammatory, proadhesive gene expression pattern in the vessel wall even in absence of leukocytes. These molecular events could provide a basis for the recruitment of leukocytes and HPC. By inhibiting the expression of such genes, rapamycin may lead to a reduced recruitment of leukocytes and HPC after vascular injury, an effect that may play a decisive role for its effectiveness in reducing restenosis. PMID- 15546960 TI - NADH oxidase signaling induces cyclooxygenase-2 expression during lipopolysaccharide stimulation in cardiomyocytes. AB - Cyclooxygenase-2 (COX-2) is induced in response to lipopolysaccharide (LPS). However, the signaling mechanisms of LPS-induced COX-2 expression in cardiomyocytes are not well understood. The aim of this study was to investigate the role of gp91(phox)-containing NADH oxidase signaling pathway in LPS-induced COX-2 expression in cardiomyocytes. Cultured neonatal mouse cardiomyocytes showed basal COX-2 expression and PGE2 production. In response to LPS, COX-2 expression and PGE2 production increased by two- to four-fold, which were completely blocked by a selective COX-2 inhibitor NS398. LPS also increased NADH oxidase (gp91(phox) and p47(phox) subunits) expression and superoxide generation. Deficiency of gp91(phox) or suppression of p22(phox) expression decreased NADH oxidase activity and down-regulated COX-2 expression and PGE2 production stimulated by LPS. Pharmacological inhibitors of NADH oxidase prevented LPS-induced COX-2 expression and PGE2 production. The effect of NADH oxidase was mediated through MAPK activation, since inhibition of NADH-oxidase activity prevented phosphorylation of ERK1/2, p38, and JNK1/2, as well as selective inhibition of each subfamily of MAPK by siRNAs and a dominant negative mutant of JNK1 decreased COX-2 expression and completely abrogated PGE2 production in response to LPS. Furthermore, LPS induced NF-kappaB activation was decreased by inhibition of NADH oxidase, ERK1/2 or JNK1/2 activation, suggesting that LPS increases NF-kappaB activity and COX-2 expression via NADH oxidase-dependent activation of ERK1/2 and JNK1/2. In conclusion, NADH oxidase signaling represents a novel pathway leading to COX-2 expression via MAPK/NF-kappaB-dependent mechanisms in cardiomyocytes during LPS stimulation. Our study suggests that gp91(phox)-containing NADH oxidase is a potential therapeutic target of sepsis. PMID- 15546961 TI - Novel interaction partners of the TPR/MET tyrosine kinase. AB - A large variety of biological processes is mediated by stimulation of the receptor tyrosine kinase MET. Screening a mouse embryo cDNA library, we were able to identify several novel, putative intracellular TPR/MET-substrates: SNAPIN, DCOHM, VAV-1, Sorting nexin 2, Death associated protein kinase 3, SMC-1, Centromeric protein C, and hTID-1. Interactions as identified by yeast two-hybrid analysis were validated in vitro and in vivo by mammalian two-hybrid studies, a far-western assay and coimmunoprecipitation. Participation in apoptosis regulating mechanisms through interaction with DAPK-3 and cell cycle control via binding to nuclear proteins such as CENPC and SMC-1 are possible new aspects of intracellular MET signaling. PMID- 15546962 TI - Aortic stiffness in systemic sclerosis is increased independently of the extent of skin involvement. AB - OBJECTIVE: To study the stiffness of large arteries in relation to the extent of skin and lung fibrosis, aortic distensibility was examined in patients with diffuse and limited systemic sclerosis (SSc). METHODS: Consecutive patients (55 with diffuse and 51 with limited SSc) without signs and symptoms of heart failure or a previous history of arterial hypertension underwent echocardiography and lung function tests. Aortic stiffness was determined non-invasively by aortic distensibility and aortic strain measurements in all patients and in 50 healthy subjects, matched for age and gender. RESULTS: Aortic distensibility in patients with either diffuse (2.03 +/- 0.26 x 10(-6) cm(2) dyn(-1)) or limited SSc (2.12 +/- 0.33) was similarly decreased compared with controls (2.49 +/- 0.36, P<0.001). Moreover, aortic strain was significantly reduced in both patient groups compared with controls, confirming that aortic stiffness is increased in SSc independently of the extent of skin involvement. Left ventricular performance was similar between patients and controls, while left ventricular mass and tricuspid systolic gradient were significantly increased in both SSc groups, the latter being associated with aortic stiffness in multivariate analysis. No association with serum levels of C-reactive protein or lung function abnormalities indicative of pulmonary fibrosis were found. CONCLUSIONS: Stiffness of the aorta is increased in patients with established SSc regardless of the extent of the inflammatory fibrotic process in the skin and lungs, suggesting that additional pathogenetic mechanisms contribute to the compromise of large arteries. PMID- 15546963 TI - A comparison of the semiflexed (MTP) view with the standing extended view (SEV) in the radiographic assessment of knee osteoarthritis in a busy routine X-ray department. AB - OBJECTIVE: To compare the reproducibility of the standing extended view (SEV) (also known as the standing anteroposterior view) with the semiflexed, postero anterior view [the 'metatarsophalangeal' (MTP)] view for assessing joint space width (JSW) and osteophytes in osteoarthritis of the knee when used in a busy routine X-ray department. METHODS: Forty-seven patients (24 men) had both SEV and MTP views taken on the same day in a busy National Health Service radiography department. Repeat views were taken as entirely separate procedures some time over the following 2 weeks, in the same department and with no special arrangements for the selection of radiographers, time of day, or X-ray machine. The first 24 patients had second views in the SEV position whilst the remaining 23 had second MTP views. Radiographs were read independently by two experienced observers who measured JSW with a transparent ruler to the nearest 0.5 mm at the narrowest point in both medial and lateral compartments of the tibiofemoral joint in both knees. Osteophytes were graded 0-2 according to a standard atlas. Ten SEV and 10 MTP radiographs selected randomly were re-read by one observer. RESULTS: Mean (95% confidence interval) JSW in the medial compartment measured on SEV radiographs was 3.54 mm (3.08, 3.99) and on MTP radiographs it was 2.80 mm (2.37, 3.23); in the lateral compartment it was 6.04 mm (5.71, 6.37) when measured on SEV radiographs and 5.47 mm (5.09, 5.85) on MTP radiographs. The estimated variances for the medial compartment were 2.0 mm2 for SEV and 0.2 mm2 for MTP (P < 0.001) and for the lateral compartment 1.4 mm2 for SEV and 0.5 mm2 for MTP (P < 0.001). The proportion of radiographs for which there was disagreement between observers regarding osteophyte grade was not statistically different between SEV and MTP views (SEV, medial 40%, lateral 44%; MTP, medial 39%, lateral 39%). CONCLUSIONS: Even when radiographs are taken in a busy National Health Service radiography department, measurement of JSW from the MTP view is more reproducible than from the SEV view, the MTP view gives a slightly lower measurement of JSW, and there is no advantage in using either view in recording osteophyte grade. We recommend the wider adoption of the MTP method. PMID- 15546964 TI - Low seroprevalence and poor specificity of antineutrophil cytoplasmic antibodies in tuberculosis. AB - OBJECTIVES: Recently published findings suggested that antineutrophil cytoplasmic antibodies (ANCA), particularly those with a cytoplasmic (C-ANCA) labelling pattern and targeting proteinase 3 (anti-PR3), might be markers of tuberculosis (TB). This is a critical issue, because C-ANCA/anti-PR3 were considered to be a highly specific hallmark of Wegener's granulomatosis or microscopic polyangiitis and because TB may clinically mimic Wegener's granulomatosis. We therefore undertook a study with the aim of investigating further the prevalence and specificity of ANCA in TB. METHODS: We evaluated serum samples from 67 patients diagnosed with culture-proven TB and 10 previously untested control samples from patients known to be ANCA positive (four Wegener's granulomatosis and two microscopic polyangiitides) or negative. All 77 sera were screened for ANCA using commercially available indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assay (ELISA) for anti-PR3 and antimyeloperoxidase (MPO). IIF positive and anti-PR3- and anti-MPO-negative sera were also tested for bactericidal/permeability-increasing protein, lactoferrin, elastase and cathepsin G specificities with commercially available ELISA. RESULTS: IIF detected ANCA in seven (10%) of the TB sera, including three C-ANCA and four atypical perinuclear labelling ANCA. Only one IIF-negative specimen was anti-PR3 positive in ELISA. ANCA testing of the control sera yielded IIF and ELISA results concordant with previous findings, except for one borderline ELISA. CONCLUSION: Our results indicate that TB is associated with low ANCA seroprevalence and poor specificity, with no test serum showing combined C-ANCA/anti-PR3 activity. In a clinical setting of Wegener's granulomatosis/TB mimicry, such combined reactivity would seem to be more suggestive of Wegener's granulomatosis. PMID- 15546965 TI - Single-nucleotide polymorphisms in the SPARC gene are not associated with susceptibility to scleroderma. AB - OBJECTIVE: SPARC (secreted protein, acidic and rich in cysteine) is a matricellular protein that modulates cell-cell and cell-extracellular matrix interactions. SPARC expression is restricted mainly to sites of tissue remodelling and wound repair, and is prominent in fibrotic disorders. Single nucleotide polymorphisms (SNPs) in the SPARC gene are reportedly linked to scleroderma in four ethnic groups: Choctaw Indians, Caucasians, African Americans and Mexican Americans. We set out to reproduce and to positionally clone these disease associations in a set of UK Caucasian scleroderma patients and ethnically matched controls. METHODS: One hundred and twenty-one scleroderma subjects and 200 controls were genotyped by polymerase chain reaction with sequence-specific primers differing only in the 3' nucleotide corresponding to each allele of the biallelic SNPs. Scleroderma patients were analysed against controls and on the basis of their fibrosing alveolitis status as judged by high-resolution computed tomography evaluation and the extent of cutaneous involvement. RESULTS: Eight biallelic SNPs were genotyped: three from the last untranslated exon, which had been described previously, and an additional five novel SNPs: two in the promoter region, one in exon three and two in the 3' untranslated region. Six major haplotypes were constructed across all eight SNP positions. No significant differences in genotype, allele or haplotype frequency were observed between scleroderma and controls or within scleroderma subgroups. CONCLUSIONS: SNPs in the SPARC gene are not associated with susceptibility to scleroderma. This research adds to the genetic knowledge of the SPARC gene by identifying five novel SNPs spanning the whole gene and inserting these within the context of clearly defined haplotypes. PMID- 15546966 TI - Lack of association of matrix metalloproteinase 3 (MMP3) genotypes with ankylosing spondylitis susceptibility and severity. AB - OBJECTIVE: To study the linkage and association of ankylosing spondylitis (AS) with genotypes for matrix metalloproteinase 3 (MMP3), a gene located at chromosome 11q22.3 and lying within the 101-124 cM region observed in a recent genome-wide scan as a region associated with AS susceptibility. METHODS: MMP3 genotypes were examined in 229 pedigrees with AS, 131 sporadic AS cases and 87 Caucasian controls. Eight single-nucleotide polymorphisms (SNPs) were selected and genotyped using Taqman. Non-parametric linkage (NPL) analysis was conducted between the eight MMP3 SNPs and AS using the NPL-all statistic and two-point parametric linkage analysis using GeneHunter Plus. Unrelated AS cases and controls were compared using chi2 statistics, and family-based controls using the transmission disequilibrium test and pedigree disequilibrium test. RESULTS: None of the eight MMP3 SNPs were significantly associated with AS, either using the 131 sporadic cases alone or in analyses which combined these cases with the 226 unrelated affected AS patients derived from the pedigrees. Analysis of linkage disequilibrium (LD) demonstrated that seven of the eight SNPs studied were in strong LD except for rs626750, which is about 6 kb upstream of the 5' end of the gene. No significant linkage was observed using NPL and LODs in the families. No association was seen of any of the MMP3 SNPs with disease severity (defined by patient functioning), as measured either by the Bath Ankylosing Spondylitis Functional Index or the modified Health Assessment Questionnaire. CONCLUSION: These data suggest that MMP3 genotypes are not involved in AS susceptibility or severity. PMID- 15546967 TI - Calcium channel blockers for primary Raynaud's phenomenon: a meta-analysis. AB - BACKGROUND: To determine the efficacy of calcium channel blockers (CCBs) for primary Raynaud's phenomenon (RP). Primary outcomes were frequency and severity of RP attacks. METHODS: A meta-analysis was conducted using primary data sources: Medline, Current Contents and the Cochrane Controlled Trials Register. Inclusion criteria were randomized controlled trials (RCTs) of >2 days' duration with <35% dropouts. Eighteen of 31 trials were eligible for inclusion [13 nifedipine vs placebo; five other CCBs vs placebo (n = 361)]. The main reasons for trial exclusion were: subset data (primary RP) not provided (n = 10); data published more than once (n = 1); lack of control group (n = 1); and lack of randomization (n = 1). Data were abstracted independently and a weighted mean difference (WMD) was calculated for the outcomes. RESULTS: The WMD (95% confidence interval) of all CCBs vs placebo for reduction in the frequency of attacks (n = 17) over 1 week was -5.00 (-9.02, -0.99) (P = 0.01) or -2.80 (-3.90, -1.70) when heterogeneity was considered, and -6.05 (-11.19,-0.19) (P = 0.04) for nifedipine alone (n = 10). The WMD of all CCBs vs placebo (n = 8) for reduction in severity of attacks (assessed with a 10-cm visual analogue scale) was -1.39 (-2.20, -0.58) (P = 0.0007) and -1.81 (-3.08, -0.54) (P = 0.005) for nifedipine alone (n = 5). CONCLUSIONS: Several small RCTs of CCBs for primary RP have been conducted and have yielded clinical improvement in the frequency and severity of ischaemic attacks. Most trials were crossover trials in which order effect was not studied; this may have introduced bias. The effect size may have been small because of low dosing in studies. The efficacy of CCBs for reducing severity and frequency of ischaemic attacks in primary RP is small (average of 2.8 to 5.0 fewer attacks per week and a 33% reduction in severity). PMID- 15546968 TI - Experimental study of teicoplanin, alone and in combination, in the therapy of cephalosporin-resistant pneumococcal meningitis. AB - OBJECTIVES: The aim of the study was to determine the efficacy of teicoplanin, alone and in combination with ceftriaxone, in a rabbit model of cephalosporin resistant pneumococcal meningitis, and to assess the effect of concomitant therapy with dexamethasone. METHODS: In vitro killing curves of teicoplanin, with and without ceftriaxone, were performed. Groups of eight animals per treatment were inoculated with a cephalosporin-resistant pneumococcal strain (penicillin MIC, 4 mg/L; ceftriaxone MIC, 2 mg/L; teicoplanin MIC, 0.03 mg/L) and treated over a 26 h period. Teicoplanin was administered at a dose of 15 mg/kg, alone and in combination with ceftriaxone at 100 mg/kg with or without dexamethasone at 0.25 mg/kg. CSF samples were collected at different time-points, and bacterial titres, white blood cell counts, lactate and protein concentrations and bacteriostatic/bactericidal titres were determined. Blood and CSF teicoplanin pharmacokinetic and pharmacodynamic parameters were determined. RESULTS: Teicoplanin alone promoted a decrease in bacterial counts at 6 h of -2.66 log cfu/mL and was bactericidal at 24 h, without therapeutic failures. Similar good results were obtained when dexamethasone was used simultaneously, in spite of the penetration of teicoplanin into the CSF being significantly reduced, from 2.31% to 0.71%. Teicoplanin and ceftriaxone combinations were synergic in vitro, but not in the meningitis model. CONCLUSIONS: Teicoplanin alone was very effective in this model of cephalosporin-resistant pneumococcal meningitis. The use of concomitant dexamethasone resulted in lower CSF teicoplanin levels, but not in therapeutic failures. The combination of teicoplanin plus ceftriaxone and dexamethasone might be a good alternative for the empirical therapy of pneumococcal meningitis. Additional data should confirm our experiments, in advance of clinical trials to assess efficacy in humans. PMID- 15546969 TI - Rapid flow-cytometric susceptibility testing of Candida species. AB - OBJECTIVES: To develop a rapid flow-cytometric antifungal susceptibility test and to compare results with the standard methods. METHODS: Reference and laboratory strains of Candida were tested for susceptibility to fluconazole and echinocandin by fluorescent flow cytometry using Acridine Orange as indicator of viability. Flow cytometry results were compared with MICs as determined by macrodilution and/or Etest. RESULTS: Seventy Candida strains were tested for susceptibility to fluconazole, and 74 strains for susceptibility to echinocandin. Minimal concentration of fluconazole causing 40% cell damage, as determined by flow cytometry, showed excellent association with MIC, as determined by other methods. The flow method, completed within 5 h, had excellent sensitivity and specificity to distinguish between sensitive, susceptible dose-dependent and resistant strains. The flow cytometry method for echinocandin was completed within 3 h, and minimal concentration causing 50% cell damage was associated with MIC as determined by macrodilution. CONCLUSIONS: Antifungal susceptibility testing by FACS is a reliable, rapid method for determining susceptibility of Candida to fluconazole and echinocandin. The method allows same-day results, assisting in the selection of appropriate antifungal therapy. PMID- 15546970 TI - Determination of antifungal drug susceptibilities of Aspergillus species by a fluorescence-based microplate assay. AB - OBJECTIVES: We have investigated the use of a viability dye, chloromethylfluorescein di-acetate (CMFDA), for antifungal susceptibility testing in a fluorescence microplate (FM) assay format. METHODS: For this FM assay, conidia were incubated in increasing concentrations of antifungal drug for 16 h and stained with CMFDA. Fluorescence, measured as mean fluorescence units (MFU) in a fluorescence microplate reader, was graphed relative to that of a drug-free control, and the MIC was defined as the lowest concentration of the drug that resulted in complete reduction (100%) in MFU for amphotericin B, or 90% reduction in MFU for itraconazole and voriconazole. Susceptibilities of 10 clinical isolates of Aspergillus fumigatus, Aspergillus terreus and Aspergillus niger to amphotericin B, itraconazole and voriconazole were tested in a blinded fashion using the FM and the NCCLS methods. RESULTS AND CONCLUSIONS: Reproducibility of the FM assay was excellent, and results correlated with those of the NCCLS microdilution method. The FM assay appears to be a rapid, objective method for testing fungal susceptibilities to itraconazole, voriconazole and amphotericin B. PMID- 15546971 TI - Nosocomial outbreak by Proteus mirabilis producing extended-spectrum beta lactamase VEB-1 in a Korean university hospital. AB - OBJECTIVES: To examine the molecular mechanisms involved in the beta-lactam resistance of multidrug-resistant Proteus mirabilis isolates that showed an unusual synergy between imipenem and ceftazidime in a Korean hospital. METHODS: Over an 11 month period, a total of 12 P. mirabilis isolates showing resistance to ampicillin, gentamicin, ceftazidime, cefotaxime, cefuroxime, cefalothin, cefepime, piperacillin, trimethoprim/sulfamethoxazole and ciprofloxacin, were recovered from the sputum and urine specimens of nine patients who were hospitalized in the neurosurgery ward. The extended-spectrum beta-lactamases were screened with a double disc synergy test using ceftazidime, cefotaxime, aztreonam, cefepime and clavulanate. The ESBL types were determined by PCR using specific primers for bla(TEM-1), bla(SHV-1), bla(CTX-M-1), bla(CTX-M-2), bla(CTX M-8), bla(CTX-M-9), bla(PER-1), bla(GES-1), bla(VEB-1), bla(OXA-10) and bla(OXA 13) followed by sequencing. All the isolates underwent molecular typing by PFGE. The transferability was examined by conjugation. RESULTS AND CONCLUSIONS: All the isolates showed a marked synergy between the extended-spectrum cephalosporins and clavulanate together with an unusual synergy between cefoxitin and the cephalosporins (cefalothin, cefuroxime, ceftazidime, cefotaxime) and between imipenem, and ceftazidime and cefotaxime. Isoelectric focusing of the crude bacterial extracts showed a beta-lactamase band with a pI value of 5.4, which was inhibited by clavulanate. PCR and sequencing identified the gene to be bla(VEB 1). In addition, the aadB gene was detected, conferring aminoglycoside resistance. The resistance was not transferred by conjugation. The outbreak was of a clonal origin as shown by PFGE demonstrating an identical banding pattern. This is the first report of VEB-1-producing Enterobacteriaceae in Korea. PMID- 15546972 TI - Antibiotic combinations for serious infections caused by carbapenem-resistant Acinetobacter baumannii in a mouse pneumonia model. AB - OBJECTIVES: Successful therapy of carbapenem-resistant Acinetobacter baumannii strains has been reported with colistin, but recently we argued against its use as monotherapy because of the poor results obtained in a mouse pneumonia model. Our aim was to identify antibiotic combinations that were valid therapeutic alternatives in the same model. METHODS: We used two carbapenem-resistant A. baumannii strains (D and E; MICs of imipenem, 8 and 512 mg/L, respectively). MICs of tobramycin, rifampicin and colistin for both strains were 8, 8 and 0.5 mg/L, respectively. RESULTS: In infections caused by strain D, lung bacterial counts (log(10) cfu/g, mean +/- s.d.) were: controls (10.86+/-0.25), imipenem (5.99+/ 0.59, P < 0.05 versus controls), and colistin (10.43 +/- 1.09); imipenem + tobramycin was the most active combination (5.46+/-0.62, P < 0.05 versus controls). In infections caused by strain E, results were: controls (10.82+/ 0.33), rifampicin (5.62+/-0.26, P < 0.05 versus controls), colistin (8.38+/-1.22, P < 0.05 versus controls), and imipenem (11.01+/-0.2); rifampicin + imipenem (3.79+/-0.99) and rifampicin + tobramycin (3.96+/-0.30) were the most active combinations (P < 0.05); results with rifampicin + colistin (5.59+/-1.17) were similar to those with rifampicin alone. CONCLUSIONS: Our data indicate that imipenem can still be the best alternative for carbapenem-resistant A. baumannii infections with moderate levels of imipenem resistance, preferably combined with aminoglycosides. For strains highly resistant to imipenem, a combination of rifampicin with imipenem, tobramycin or colistin may be useful, if resistance to rifampicin is only moderate. PMID- 15546973 TI - Antibiotic prescribing for respiratory tract infections in Dutch primary care in relation to patient age and clinical entities. AB - OBJECTIVES: To obtain detailed information on current prescribing rates of antibiotics for respiratory tract infections (RTIs) in Dutch general practice and its relation with age and respiratory tract clinical entities. METHODS: We assessed the mean proportion of antibiotics prescribed for RTIs per age group, contact-based and population-based using all patient contacts concerning RTIs in the year 2000 selected from the IPCI database, containing information on general practice consultations of 235,290 patients. RESULTS: In one-third of all contacts concerning RTIs, antibiotics were prescribed, with much variation between age groups and clinical entities. For children (0-15 years) and the elderly (over 75 years), the lowest contact-based percentages of prescribed antibiotics for RTIs were found, while population-based, children of age 0-5 years received far more antibiotics for RTIs. High prescribing rates were seen in patients with sinusitis like complaints (67%) or pneumonia (78%), whereas low rates were found for patients with upper RTIs (16%). CONCLUSIONS: Potential over-prescribing of antibiotics for RTIs occurs in the age group 31-65 years, not in children and the elderly, and in patients with upper RTIs, sinusitis and most likely acute bronchitis (contact-based). The management of these subgroups of patients should be addressed in quality assurance programmes. Children and the elderly visit the GP much more often than adults, which can be explained by more frequent (children) or more severe (elderly) RTI morbidity, but in proportion they do not receive more antibiotics. PMID- 15546974 TI - Guidelines for the antibiotic treatment of endocarditis in adults: report of the Working Party of the British Society for Antimicrobial Chemotherapy. AB - The BSAC Guidelines on Endocarditis were last published in 1998. The Guidelines presented here have been updated and extended to reflect changes in both the antibiotic resistance characteristics of causative organisms and the availability of new antibiotics. Randomized, controlled trials suitable for the development of evidenced-based guidelines in this area are still lacking, and therefore a consensus approach has again been adopted. The Guidelines cover diagnosis and laboratory testing, suitable antibiotic regimens and causative organisms. Special emphasis is placed on common causes of endocarditis, such as streptococci and staphylococci, however, other bacterial causes (such as enterococci, HACEK organisms, Coxiella and Bartonella) and fungi are considered. The special circumstances of prosthetic endocarditis are discussed. PMID- 15546975 TI - Interspecies communication in Streptococcus gordonii-Veillonella atypica biofilms: signaling in flow conditions requires juxtaposition. AB - During the development of human oral biofilm communities, the spatial arrangement of the bacteria is thought to be driven by metabolic interactions between them. Streptococcus gordonii and Veillonella atypica, two early colonizing members of the dental plaque biofilm, have been postulated to participate in metabolic communication; S. gordonii ferments carbohydrates to form lactic acid, which is a preferred fermentation substrate for V. atypica. We found that, during agar-plate coculture of these organisms, a signaling event occurs that results in increased expression of the S. gordonii alpha-amylase-encoding gene amyB. Confocal scanning laser microscopy of coculture flowcell-grown biofilms using human saliva as the sole nutrient showed that V. atypica caused S. gordonii to increase expression of a PamyB-'gfp transcriptional fusion in a spatially resolved fashion. In this open system, only those streptococci in mixed-species microcolonies containing V. atypica expressed GFP; nearby S. gordonii colonies that lacked V. atypica did not express GFP. In a closed system containing S. gordonii and V. atypica, flow cytometric analysis showed that S. gordonii containing the PamyB-'gfp reporter plasmid exhibited mean fluorescence levels 20-fold higher than did S. gordonii that had not been incubated with V. atypica. Thus, in a closed system where a diffusible signal can accumulate above a required threshold, interspecies signaling mediates a change in gene expression. We provide evidence that, in open systems like those that predominate in natural biofilms, diffusible signals between species are designed to function over short distances, on the order of 1 mum. PMID- 15546976 TI - Unraveling the interface of signal recognition particle and its receptor by using chemical cross-linking and tandem mass spectrometry. AB - Among the methods used to unravel protein interaction surfaces, chemical cross linking followed by identification of the cross-linked peptides by mass spectrometry has proven especially useful in dynamic and complex systems. During the signal recognition particle (SRP)-dependent targeting of proteins to the bacterial plasma membrane, the specific interaction between Ffh (the protein component of SRP) and FtsY (the SRP receptor) is known to be essential for the efficiency and fidelity of this process. In this work, we studied the Escherichia coli and Thermus aquaticus Ffh.FtsY complexes by using chemical cross-linking and tandem mass spectrometry to identify nine intermolecular cross-linked peptides. This information was used in conjunction with a previously undescribed model building approach that combines geometric restraint optimization with macromolecular docking. The resulting model of the Ffh.FtsY complex is in good agreement with the crystal structure solved shortly thereafter. Intriguingly, four of the cross-linked pairs involve the M domain of Ffh, which is absent from the crystal structure, providing previously undocumented experimental evidence that the M domain is positioned in close proximity to the Ffh.FtsY interface in the complex. PMID- 15546977 TI - Getting protein solvent structures down cold. PMID- 15546978 TI - An atomic-resolution mechanism of 3-hydroxy-3-methylglutaryl-CoA synthase. PMID- 15546979 TI - Synaptotagmin promotes both vesicle fusion and recycling. PMID- 15546980 TI - Germ-line immortality. PMID- 15546981 TI - A mitotic kinesin-like protein required for normal karyokinesis, myosin localization to the furrow, and cytokinesis in Dictyostelium. AB - Dictyostelium mitotic kinesin Kif12 is required for cytokinesis. Myosin II localization to the cleavage furrow is severely depressed in Kif12-null (Deltakif12) cells, which accounts in part for the cytokinesis failure. Myosin II null cells, however, undergo mitosis-coupled cytokinesis when adhering to a surface, whereas the Deltakif12 cells cannot. During mitosis, the rate of change of internuclear separation in Deltakif12 cells is reduced compared with wild-type cells, indicating multiple roles of this molecular motor during mitosis and cytokinesis. GFP-Kif12, which rescues wild-type behavior when expressed in the Deltakif12 strain, is concentrated in the nucleus in interphase cells, translocates to the cytoplasm at the onset of mitosis, appears in the centrosomes and spindle, and later is concentrated in the spindle midbody. Given these results, we hypothesize a mechanism for myosin II translocation to the furrow to set up the contractile ring. PMID- 15546982 TI - Araceae from the Early Cretaceous of Portugal: evidence on the emergence of monocotyledons. AB - A new species (Mayoa portugallica genus novum species novum) of highly characteristic inaperturate, striate fossil pollen is described from the Early Cretaceous (Barremian-Aptian) of Torres Vedras in the Western Portuguese Basin. Based on comparison with extant taxa, Mayoa is assigned to the tribe Spathiphylleae (subfamily Monsteroideae) of the extant monocotyledonous family Araceae. Recognition of Araceae in the Early Cretaceous is consistent with the position of this family and other Alismatales as the sister group to all other monocots except Acorus. The early occurrence is also consistent with the position of Spathiphylleae with respect to the bulk of aroid diversity. Mayoa occurs in the earliest fossil floras (from circa 110 to 120 million years ago) that contain angiosperm flowers, carpels, and stamens. The new fossil provides unequivocal evidence of monocots in early angiosperm assemblages that also include a variety of key "magnoliid" lineages (e.g., Chloranthaceae) but only a limited diversity of eudicots. PMID- 15546984 TI - Coding sequences of functioning human genes derived entirely from mobile element sequences. AB - Among all of the many examples of mobile elements or "parasitic sequences" that affect the function of the human genome, this paper describes several examples of functioning genes whose sequences have been almost completely derived from mobile elements. There are many examples where the synthetic coding sequences of observed mRNA sequences are made up of mobile element sequences, to an extent of 80% or more of the length of the coding sequences. In the examples described here, the genes have named functions, and some of these functions have been studied. It appears that each of the functioning genes was originally formed from mobile elements and that in some process of molecular evolution a coding sequence was derived that could be translated into a protein that is of some importance to human biology. In one case (AD7C), the coding sequence is 99% made up of a cluster of Alu sequences. In another example, the gene BNIP3 coding sequence is 97% made up of sequences from an apparent human endogenous retrovirus. The Syncytin gene coding sequence appears to be made from an endogenous retrovirus envelope gene. PMID- 15546983 TI - Fear fosters flight: a mechanism for fear contagion when perceiving emotion expressed by a whole body. AB - Darwin regarded emotions as predispositions to act adaptively, thereby suggesting that characteristic body movements are associated with each emotional state. To this date, investigations of emotional cognition have predominantly concentrated on processes associated with viewing facial expressions. However, expressive body movements may be just as important for understanding the neurobiology of emotional behavior. Here, we used functional MRI to clarify how the brain recognizes happiness or fear expressed by a whole body. Our results indicate that observing fearful body expressions produces increased activity in brain areas narrowly associated with emotional processes and that this emotion-related activity occurs together with activation of areas linked with representation of action and movement. The mechanism of fear contagion hereby suggested may automatically prepare the brain for action. PMID- 15546985 TI - Partitions with difference conditions and Alder's conjecture. AB - In 1956, Alder conjectured that the number of partitions of n into parts differing by at least d is greater than or equal to that of partitions of n into parts identical with +/-1 (mod d+3) for d > or = 4. In 1971, Andrews proved that the conjecture holds for d = 2r-1, r > or = 4. We sketch a proof of the conjecture for all d > or = 32. PMID- 15546986 TI - Defining the function of a prolactin gene family member. PMID- 15546987 TI - Ultraviolet plumage reflectance distinguishes sibling bird species. AB - Realistic studies of plumage color need to consider that many birds can see near UV light, which normal humans cannot perceive. Although previous investigations have revealed that UV-based plumage reflectance is an important component of various intraspecific social signals, the contribution of UV signals to inter specific divergence and speciation in birds remains largely unexplored. I describe an avian example of an interspecific phenomenon in which related sympatric species that appear similar to humans (sibling species) differ dramatically in the UV. Both UV video images and physical reflectance spectra indicate that the dorsal plumage of the tanager Anisognathus notabilis has a strong UV-limited reflectance band that readily distinguishes this species from its sibling congener Anisognathus flavinuchus. The main human-visible distinction between A. notabilis (olive back) and coexisting A. flavinuchus (black back) also occurs among different geographic populations of A. flavinuchus. Notably, however, olive- and black-backed taxa interbreed (differentiated populations of A. flavinuchus) unless the additional UV distinction is present (A. notabilis vs. A. flavinuchus). Thus, UV-based reflectance can be an essential component of plumage divergence that relates to reproductive isolation, a key attribute of biological species. PMID- 15546988 TI - Neural synchrony indexes disordered perception and cognition in schizophrenia. AB - Current views of schizophrenia suggest that it results from abnormalities in neural circuitry, but empirical evidence in the millisecond range of neural activity has been difficult to obtain. In pursuit of relevant evidence, we previously demonstrated that schizophrenia is associated with abnormal patterns of stimulus-evoked phaselocking of the electroencephalogram in the gamma band (30 100 Hz). These patterns may reflect impairments in neural assemblies, which have been proposed to use gamma-band oscillations as a mechanism for synchronization. Here, we report the unique finding that, in both healthy controls and schizophrenia patients, visual Gestalt stimuli elicit a gamma-band oscillation that is phase-locked to reaction time and hence may reflect processes leading to conscious perception of the stimuli. However, the frequency of this oscillation is lower in schizophrenics than in healthy individuals. This finding suggests that, although synchronization must occur for perception of the Gestalt, it occurs at a lower frequency because of a reduced capability of neural networks to support high-frequency synchronization in the brain of schizophrenics. Furthermore, the degree of phase locking of this oscillation is correlated with visual hallucinations, thought disorder, and disorganization in the schizophrenia patients. These data provide support for linking dysfunctional neural circuitry and the core symptoms of schizophrenia. PMID- 15546989 TI - NO and superoxide: opposite ends of the seesaw in cardiac contractility. PMID- 15546991 TI - On the mechanism and biology of cytochrome oxidase inhibition by nitric oxide. AB - The detailed molecular mechanism for the reversible inhibition of mitochondrial respiration by NO has puzzled investigators: The rate constants for the binding of NO and O2 to the reduced binuclear center CuB/a3 of cytochrome oxidase (COX) are similar, and NO is able to dissociate slowly from this center whereas O2 is kinetically trapped, which altogether seems to favor the complex of COX with O2 over the complex of COX with NO. Paradoxically, the inhibition of COX by NO is observed at high ratios of O2 to NO (in the 40-500 range) and is very fast (seconds or faster). In this work, we used simple mathematical models to investigate this paradox and other important biological questions concerning the inhibition of COX by NO. The results showed that all known features of the inhibition of COX by NO can be accounted for by a direct competition between NO and O2 for the reduced binuclear center CuB/a3 of COX. Besides conciliating apparently contradictory data, this work provided an explanation for the so called excess capacity of COX by showing that the COX activity found in tissues actually is optimized to avoid an excessive inhibition of mitochondrial respiration by NO, allowing a moderate, but not excessive, overlap between the roles of NO in COX inhibition and in cellular signaling. In pathological situations such as COX-deficiency diseases and chronic inflammation, an excessive inhibition of the mitochondrial respiration is predicted. PMID- 15546990 TI - Reduced cGMP signaling associated with neointimal proliferation and vascular dysfunction in late-stage atherosclerosis. AB - Atherosclerosis is associated with alterations in nitric oxide (NO)/cGMP signaling. In early stages of the disease, inflammatory and possibly other cells produce reactive oxygen species that scavenge vasoprotective NO. In addition to the oxidative stress, expression and activity of enzymes downstream to NO formation may also be affected. Here, we show in the aortas of chronically hypercholesterolemic rabbits (a model of late-stage atherosclerosis), both subunits and specific activity of the NO receptor soluble guanylyl cyclase (sGC) were significantly reduced, whereas overall NO synthase activity was unaffected. These changes were most prominent in the neointimal layer, wherein cGMP-dependent protein kinase I (cGK) levels also were reduced. Additionally, a protein (p38(nt)) that was constitutively tyrosine-nitrated was detected, and its expression was significantly reduced in atherosclerotic aorta. Phosphorylation of the cGK substrate vasodilator-stimulated phosphoprotein (VASP) at Ser-239, an established biochemical endpoint of NO/cGMP signaling, also was reduced. Thus, late-stage atherosclerosis is associated not only with enhanced NO breakdown but also with altered NO reception and cGMP signaling. Preferential down-regulation in neointima suggests a direct connection of these changes to neointimal proliferation and vascular dysfunction and provides a rationale for future pharmacotherapy using classical and novel sGC activators. PMID- 15546992 TI - Trans-bonded pairs of E-cadherin exhibit a remarkable hierarchy of mechanical strengths. AB - Classical cadherins are primary mediators of calcium-dependent cell interactions in multicellular organisms. Organized in five tandemly repeated E-cadherin (EC) modules, the extracellular segments of these membrane-spanning glycoproteins interact homophilically between opposing cells to create highly regulated patterns of attachment stabilized by cytoskeletal elements inside the cells. Despite many structural and functional studies, a significant controversy exists in regard to the organization of cadherin binding in adhesion sites. Supported by considerable evidence, perhaps the most widely held view is that opposing N terminal EC1-EC2 (EC12) domains form a "zipper" of bonds. However, immobilized on two atomically smooth surfaces and pushed to adhesive contact, opposing cadherins become fully interdigitated and unbind through three discrete jumps comparable with domain dimensions when pulled apart. So the question remains as to whether mechanical adhesion strength emanates solely from interactions between the peripheral N-terminal domains or involves multiple overlapping domains. It is also unclear whether a primary adhesion complex is formed by a single opposing pair of cadherins or whether the complex involves a more complicated network of cis-bonded multimers. To address these questions, we used a special jump/ramp mode of force spectroscopy to test isolated pairwise interactions between recombinant fragments of ECs. Besides the formation of strong trans-bonded dimers, we find a remarkable hierarchy of rupture strengths for bonds between the full five-domain fragments that suggests multiple mechanical functions for cadherins, perhaps providing distinct properties needed for transient-specific recognition as well as stable tissue formation. PMID- 15546993 TI - Glyceraldehyde 3-phosphate dehydrogenase-S, a sperm-specific glycolytic enzyme, is required for sperm motility and male fertility. AB - Although glycolysis is highly conserved, it is remarkable that several unique isozymes in this central metabolic pathway are found in mammalian sperm. Glyceraldehyde 3-phosphate dehydrogenase-S (GAPDS) is the product of a mouse gene expressed only during spermatogenesis and, like its human ortholog (GAPD2), is the sole GAPDH isozyme in sperm. It is tightly bound to the fibrous sheath, a cytoskeletal structure that extends most of the length of the sperm flagellum. We disrupted Gapds expression by gene targeting to selectively block sperm glycolysis and assess its relative importance for in vivo sperm function. Gapds( /-) males were infertile and had profound defects in sperm motility, exhibiting sluggish movement without forward progression. Although mitochondrial oxygen consumption was unchanged, sperm from Gapds(-/-) mice had ATP levels that were only 10.4% of those in sperm from WT mice. These results imply that most of the energy required for sperm motility is generated by glycolysis rather than oxidative phosphorylation. Furthermore, the critical role of glycolysis in sperm and its dependence on this sperm-specific enzyme suggest that GAPDS is a potential contraceptive target, and that mutations or environmental agents that disrupt its activity could lead to male infertility. PMID- 15546994 TI - Distinct pathways of insulin-regulated versus diabetes-regulated gene expression: an in vivo analysis in MIRKO mice. AB - Diabetes mellitus is a complex metabolic disorder accompanied by alterations in cellular physiology, metabolism, and gene expression. These alterations can be primary (due to loss of direct insulin action) or secondary (due to the metabolic perturbations associated with the disease). To dissect and quantitate these two separate effects, we compared the skeletal muscle gene-expression profiles of muscle insulin receptor knockout (MIRKO) mice and their Lox controls in the basal, streptozotocin-induced diabetic, and insulin-treated diabetic states. Pure deficiency of insulin action as present in the MIRKO mouse results in regulation of 130 genes, with down-regulation of NSF (N-ethylmaleimide-sensitive fusion protein) and VAMP-2 (vesicle-associated membrane protein 2), stearoyl CoA desaturase 1, and cAMP-specific phosphodiesterase 4B, as well as up-regulation of some signaling-related genes, such as Akt2, and the fatty-acid transporter CD36. In diabetes, additional transcriptional mechanisms are activated, resulting in alterations in expression of approximately 500 genes, including a highly coordinated down-regulation of genes of the mitochondrial electron-transport chain and one of the mammalian homologues of the histone deacetylase Sir2, which has been implicated in the link between nutrition and longevity. These distinct pathways of direct and indirect regulation of gene expression provide insights into the complex mechanisms of transcriptional control in diabetes and areas of potential therapeutic targeting. PMID- 15546995 TI - Alpha,beta hybrid peptides: a polypeptide helix with a central segment containing two consecutive beta-amino acid residues. AB - Conformational studies on the synthetic 11-aa peptide t-butoxycarbonyl (Boc)-Val Ala-Phe-alpha-aminoisobutyric acid (Aib)-(R)-beta3-homovaline (betaVal)-(S)-beta3 homophenylalanine (betaPhe)-Aib-Val-Ala-Phe-Aib-methyl ester (OMe) (peptide 1; betaVal and betaPhe are beta amino acids generated by homologation of the corresponding l-residues) establish that insertion of two consecutive beta residues into a polypeptide helix can be accomplished without significant structural distortion. Crystal-structure analysis reveals a continuous helical conformation encompassing the segment of residues 2-10 of peptide 1. At the site of insertion of the betabeta segment, helical hydrogen-bonded rings are expanded. A C15 hydrogen bond for the alphabetabeta segment and two C14 hydrogen bonds for the alphaalphabeta or betaalphaalpha segments have been characterized. The following conformational angles were determined from the crystal structure for the beta residues: betaVal-5 (= -126 degrees, = 76 degrees, and psi = -124) and betaPhe-6 (=-88 degrees, = 80 degrees, and psi =-118). The N terminus of the peptide is partially unfolded in crystals. The 500-MHz 1H-NMR studies establish a continuous helix over the entire length of the peptide in CDCl3 solution, as evidenced by diagnostic nuclear Overhauser effects. The presence of seven intramolecular hydrogen bonds is also established by using solvent dependence of NH chemical shifts. PMID- 15546996 TI - Impaired adrenal stress response in Toll-like receptor 2-deficient mice. AB - Septicemia is one of the major health concerns worldwide, and rapid activation of adrenal steroid release is a key event in the organism's first line of defense during this form of severe illness. The family of Toll-like receptors (TLRs) is critical in the early immune response upon bacterial infection, and TLR polymorphisms are frequent in humans. Here, we demonstrate that TLR-2 deficiency in mice is associated with reduced plasma corticosterone levels and marked cellular alterations in adrenocortical tissue. TLR-2-deficient mice have an impaired adrenal corticosterone release after inflammatory stress induced by bacterial cell wall compounds. This defect appears to be mediated by a decrease in systemic and intraadrenal cytokine expression, including IL-1, tumor necrosis factor alpha, and IL-6. Our data demonstrate a link between the innate immune system and the endocrine stress response. The critical role of TLR-2 in adrenal glucocorticoid regulation needs to be considered in patients with inflammatory disease. PMID- 15546997 TI - Sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) gene silencing and remodeling of the Ca2+ signaling mechanism in cardiac myocytes. AB - Transient elevations of cytosolic Ca2+ are a common mechanism of cellular signaling. In striated muscle, the sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA) plays an important role in terminating Ca2+ transients by returning cytosolic Ca2+ to intracellular stores. Stored Ca2+ can then be released again for subsequent signaling. We down-regulated SERCA2 gene expression in cultured cardiac myocytes by means of endogenous transcription of small interfering RNA encoded by an exogenous cDNA template. The cDNA template was delivered by adenovirus vector. Reduction of SERCA expression in all myocytes in culture was documented by immunochemistry, real-time RT-PCR, and determination of ATP dependent Ca2+ transport. The reduction of SERCA2 expression was associated with the up-regulation of transient receptor potential (TRP) channel proteins (TRPC4 and TRPC5) and Na+/Ca2+ exchanger, indicating that intracellular store deficiency was compensated for by Ca2+ fluxes through the plasma membrane. In fact, SERCA silencing was followed by increased transcription of Na+/Ca2+ exchanger, TRPC4, TRPC5, and related transcriptional factors, such as stimulating protein 1, myocyte enhancer factor 2, and nuclear factor of activated cells 4, through activation of calcineurin. This finding demonstrates that the observed compensation occurs through transcriptional crosstalk and the remodeling of Ca2+ signaling pathways. The wide significance of this regulatory mechanism is related to its general involvement in Ca2+ signaling dynamics and in cardiac development and hypertrophy. PMID- 15546999 TI - Altered dystrophin expression in the right atrium of a patient after Fontan procedure with atrial flutter. AB - Underlying mechanisms in the development of atrial flutter or intra-atrial re entry tachycardia in patients with structural cardiac abnormalities remain poorly defined. The right atrial myocardium from two patients with congenital heart disease was evaluated, of whom one presented with severe right atrial dilation and arrhythmia and the other with a normal right atrium, to assess whether increased right atrial pressure and volume overload give rise to sarcolemmal alteration. N-terminus dystrophin staining in the atrium from the patient who had undergone a Fontan procedure showed a normally distributed but significantly reduced staining signal compared with the second patient. This is the first report that patients with severe right atrial dilation and atrial flutter have marked reduction in atrial dystrophin expression. PMID- 15546998 TI - Self-generated diversity produces "insurance effects" in biofilm communities. AB - Diversity generally protects communities from unstable environmental conditions. This principle, known as the "insurance hypothesis," has been tested in many different ecosystems. Here we show that the opportunistic pathogen Pseudomonas aeruginosa undergoes extensive genetic diversification during short-term growth in biofilm communities. The induced genetic changes are produced by a recA dependent mechanism and affect multiple traits, including the behavior of the bacteria in biofilms. Some biofilm-derived variants exhibit an increased ability to disseminate, whereas others manifest accelerated biofilm formation. Furthermore, the presence of these functionally diverse bacteria increases the ability of biofilms to resist an environmental stress. These findings suggest that self-generated diversity in biofilms provides a form of biological insurance that can safeguard the community in the face of adverse conditions. PMID- 15547001 TI - Apical ballooning in relatives. AB - Apical ballooning of the left ventricle was first introduced as takotsubo-like left ventricular dysfunction in 1990 by Satoh and colleagues. The syndrome is characterised by reversible extensive akinesia of the apical and mid-portions of the left ventricle with hypercontraction of the basal segment. For the first time two sisters with this syndrome are reported, suggesting a possible genetic aetiology. PMID- 15547000 TI - Lipomatous hypertrophy of the interatrial septum: a commonly misdiagnosed mass often leading to unnecessary cardiac surgery. AB - Lipomatous hypertrophy of the interatrial septum is a benign cardiac mass that should be considered as part of the differential diagnosis for any atrial cardiac tumour. In the reported case, this lesion was initially suspected to be malignant and the patient was thus referred directly to cardiac surgeons for surgical removal. Unnecessary surgical intervention was swiftly averted because the cardiac surgeon promptly referred the patient for an expert echocardiogram that confirmed the diagnosis of lipomatous hypertrophy. The authors discuss the characteristic features of this lesion and how the diagnosis may be made based on several non-invasive imaging modalities without the need for a tissue biopsy. This condition is more common than initially thought and remains under-recognised by most clinicians. In such cases an increased awareness of this lesion along with the opinion of a specialist echocardiologist would help to avoid a misdiagnosis and unnecessary intervention. PMID- 15547003 TI - Interplay of diabetes and coronary heart disease on cardiovascular mortality. AB - Patients with both diabetes mellitus and prior myocardial infarction are at particularly high risk for cardiovascular mortality. PMID- 15547005 TI - Coconut atrium following thoracic radiotherapy. PMID- 15547004 TI - Cardioversion and the use of sedation. AB - Nurse led cardioversion services have achieved significant reductions in both cost and waiting time. However, the question of safety of the procedure raises several areas of concern. PMID- 15547006 TI - The effect of not using an internal mammary artery as a conduit for coronary artery bypass grafting. AB - When presented with a high risk case for coronary bypass surgery, the surgeon can confidently use an internal mammary artery as a conduit for that patient. PMID- 15547008 TI - Pathologic assessment of the vulnerable human coronary plaque. PMID- 15547007 TI - Insulin resistance as a contributor to myocardial ischaemia independent of obstructive coronary atheroma: a role for insulin sensitisation? AB - There is good evidence to suggest that insulin resistance and its surrogate markers are at least modest independent cardiovascular risk factors. However, as well as long term effects on atheromatous coronary disease, there is a well described correlation between markers of insulin resistance and endothelial dysfunction. In this review, the evidence for a relation between endothelial dysfunction and myocardial ischaemia is summarised. The evidence for a correlation between insulin resistance and endothelial dysfunction and the proposed cellular mechanisms are also examined. Finally, the potential role for insulin sensitising strategies is looked at and recent data examining their effects on both endothelial function and clinical symptoms is examined. In conclusion, it was found that insulin sensitising modalities have a potential role in the amelioration of angina and that randomised controlled studies are therefore warranted. PMID- 15547009 TI - Plaque stabilisation by systemic and local drug administration. PMID- 15547010 TI - Plaque sealing by coronary angioplasty. PMID- 15547011 TI - Cardiac papillary fibroelastoma of the mitral valve chordae. PMID- 15547012 TI - Cardiovascular disease incidence and mortality in older men with diabetes and in men with coronary heart disease. AB - OBJECTIVE: To examine the relation of diabetes and coronary heart disease (CHD; myocardial infarction (MI) or angina) to the incidence of major CHD and stroke events and total mortality. METHODS: Prospective study of 5934 men aged 52-74 years followed up for 10 years. The men were divided into five groups according to their diabetes and CHD status. RESULTS: During the follow up there were 662 major CHD events, 305 major stroke events, and 1357 deaths from all causes (637 cardiovascular disease (CVD) deaths, 417 CHD deaths). Men with diabetes had significantly increased cardiovascular and total mortality risk compared with non diabetic men with no CHD but lower risk than men with prior MI only. The adjusted relative risk for CHD deaths was 2.82 (95% confidence interval (CI) 1.85 to 4.28) in men with diabetes only, 2.12 (95% CI 1.53 to 2.93) in men with angina only, 3.91 (95% CI 3.07 to 4.99) in men with MI, and 8.93 (95% CI 6.13 to 12.99) in men with both diabetes and CHD. Case fatality among men with diabetes only was similar to those with prior MI only. CHD and CVD mortality increased with increasing duration of diabetes with risk eventually approaching that of patients with MI without diabetes. CONCLUSION: Men with diabetes only have a CVD risk intermediate between men with angina and men with prior MI. Their absolute risk is high and the prognosis for diabetic patients who develop CHD is extremely poor. PMID- 15547013 TI - Management and in-hospital outcome of patients with acute myocardial infarction admitted to intensive care units at the turn of the century: results from the French nationwide USIC 2000 registry. AB - OBJECTIVE: To assess actual practices and in-hospital outcome of patients with acute myocardial infarction on a nationwide scale. METHODS: Of 443 intensive care units in France, 369 (83%) prospectively collected data on all cases of infarction (within < 48 hours of symptom onset) in November 2000. RESULTS: 2320 patients (median age 68 years, 73% men) were included, of whom 83% had ST segment elevation infarction (STEMI). Patients without STEMI were older and had a more frequent history of cardiovascular disease. Median time to admission was 5.0 hours for patients with and 6.5 hours for those without STEMI. Reperfusion therapy was used for 53% of patients with STEMI (thrombolysis 28%, primary angioplasty 25%). In-hospital mortality was 8.7% (5.5% of patients without and 9.3% of those with STEMI). Multivariate analysis found that age, Killip class, lower blood pressure, higher heart rate on admission, anterior location of infarct, STEMI, diabetes mellitus, previous stroke, and no current smoking independently predicted in-hospital mortality. At hospital discharge, 95% received antiplatelet agents, 75% received beta blockers, and over 60% received statins. Angiotensin converting enzyme inhibitors were prescribed for 40% of the patients without and 52% of those with ST elevation. CONCLUSIONS: This nationwide registry, including all types of centres irrespective of their size and experience, shows continued improvement in patient care and outcomes. Time from symptom onset to admission, however, has not improved in recent years and reperfusion therapy is used for just over 50% of patients with STEMI, with an increasing use of primary angioplasty. PMID- 15547014 TI - An isolated single coronary artery supplying the entire myocardium in a patient with congenitally corrected transposition of the great vessels. PMID- 15547015 TI - Reverse remodelling of systolic left ventricular contraction pattern by long term cardiac resynchronisation therapy: colour Doppler shows resynchronisation. AB - OBJECTIVE: To quantify long term effects of cardiac resynchronisation therapy (CRT) by biventricular pacing in patients with heart failure (HF). METHODS: Regional changes in left ventricular (LV) contraction patterns effected by CRT in 19 patients with HF (12 with ischaemia; mean (SD) age 66 (9) years) with bundle branch block were examined by colour Doppler tissue velocity imaging (c-TVI). Time differences during main systolic tissue velocity peak (SYS) were compared in the basal and mid LV interventricular septum and in the corresponding LV free wall segments. RESULTS: From baseline to long term (9.8 (3.0) months) CRT, ejection fraction increased from 21.8 (5.4)% to 30.8 (7.6)%, LV end diastolic diameter decreased from 7.6 (0.9) cm to 7.1 (0.8) cm, and end systolic diameter decreased from 6.4 (1.2) cm to 6.0 (1.2) cm (p < 0.05). LV peak tissue velocities were unchanged during follow up. At baseline, SYS in LV free wall was typically delayed by an average of 29 ms in the basal LV site and by 18 ms in the mid LV site. The regional movements of the LV free wall and interventricular septum were separated by an average of only 14 ms and -4 ms (p < 0.05) at the basal site and by -21 ms and -16 ms at the mid LV site during short term and long term CRT, respectively. CONCLUSIONS: The improved haemodynamic functions observed during CRT may be explained by a significant resynchronisation of the regional LV movement pattern during long term follow up. PMID- 15547017 TI - Diagnostic and prognostic value of ST segment depression limited to the recovery phase of exercise stress test. AB - OBJECTIVES: To investigate the diagnostic and prognostic value of ST segment depression limited to the recovery phase of an exercise stress test, as compared with that of ST segment depression appearing during exercise. SETTING: Exercise stress test laboratory of a university hospital. PATIENTS AND DESIGN: Clinical and angiographic data were compared for 574 consecutive patients who developed ST segment depression during the active phase of an exercise test (group 1) and for 79 patients who developed ST segment depression only during the recovery phase of the exercise test (group 2). RESULTS: There were no differences between the two groups in major clinical features. Significant coronary artery stenoses were found in 488 group 1 patients (85%) and in 62 group 2 patients (78%, p = 0.14). Three vessel or left main disease was found in 166 (29%) group 1 and in 14 (18%) group 2 patients (p = 0.045). At a median follow up of 55.3 months of 321 group 1 and 54 group 2 patients, there were no significant differences in major cardiac events between the groups (univariate relative risk 0.81, 95% confidence interval 0.25 to 2.68, p = 0.72). CONCLUSION: The diagnostic and prognostic power of ST segment depression limited to the recovery phase of an exercise test is largely similar to that of ST segment depression induced during effort; thus, assessing ST segment depression during recovery can significantly improve the clinical information derived from exercise stress tests. PMID- 15547018 TI - "Myxomatous" mitral stenosis. PMID- 15547019 TI - Is echocardiography a valid tool to screen for left ventricular systolic dysfunction in chronic survivors of acute myocardial infarction? A comparison with radionuclide ventriculography. AB - OBJECTIVE: To assess the accuracy of echocardiography with Simpson's apical biplane method in screening for left ventricular systolic dysfunction (LVSD) in patients six months after acute myocardial infarction (AMI) as compared with radionuclide ventriculography by assessing the proportion of clinically significant errors that occur with echocardiography. DESIGN: Comparison of results of echocardiography and radionuclide ventriculography in assessing left ventricular ejection fraction among patients six months after AMI. SETTING: District general hospital. PATIENTS: 86 patients thrombolysed for AMI at six month follow up. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Correlation coefficients, mean differences, 95% limits of agreement, and differences of clinical significance between left ventricular ejection fraction on echocardiography and on radionuclide ventriculography. RESULTS: The correlation coefficient between techniques was 0.90, mean difference 1% (p = 0.04), and 95% limits of agreement -13.0% to 10.3%. Only one patient (1.2%, 0.0% to 6.3%) was classified as having normal systolic function on one imaging modality but significant LVSD on the other. Overall accuracy between the two techniques was 86%, kappa value of agreement 0.78. CONCLUSION: Echocardiography is a valid tool to screen for LVSD in patients six months after AMI, accurately differentiating normal from abnormal systolic function and showing excellent agreement with radionuclide ventriculography. This study supports the use of echocardiography in screening for LVSD in chronic stable patients after AMI or alternative high risk patients, with few differences of major clinical significance likely to occur. PMID- 15547020 TI - Impact of nicorandil in angina: subgroup analyses. AB - AIMS: IONA (impact of nicorandil in angina) is a randomised, double blind, placebo controlled trial of nicorandil, with a target dose of 20 mg twice daily. The consistency of benefits seen in subgroups is reported. METHODS: The primary composite end point of the study was coronary heart disease death, non-fatal myocardial infarction, or unplanned hospitalisation for cardiac chest pain. Subgroups were defined using baseline characteristics including, age, sex, histories of smoking, diabetes, hypertension, myocardial infarction, revascularisation, anginal status, anti-anginal treatment, other cardiovascular drugs, and an overall assessment of risk. RESULTS: A total of 5126 patients were randomised to receive nicorandil or identical placebo in addition to standard anti-anginal treatment. Overall, nicorandil reduced the incidence of the primary end point from 15.5% to 13.1% (hazard ratio (HR) 0.83, 95% confidence interval (CI) 0.72 to 0.97; p = 0.014). There was no evidence of significant heterogeneity of benefit across all subgroups studied. The absolute risk reduction was greatest and the numbers needed to treat to prevent one event was lowest in subjects at greatest risk. CONCLUSIONS: The IONA study demonstrates a significant improvement in outcome by nicorandil treatment across a broad range of patients with stable angina. PMID- 15547021 TI - Giant left main coronary artery aneurysm with mitral regurgitation. PMID- 15547022 TI - Established and emerging coronary risk factors in patients with heterozygous familial hypercholesterolaemia. AB - OBJECTIVES: To assess the clinical and biochemical factors associated with inter individual variation in susceptibility to coronary artery disease (CAD) in treated heterozygous familial hypercholesterolaemia. DESIGN: A cross sectional study was conducted of 410 patients recruited from six lipid clinics in the UK. RESULTS: CAD was documented in 104 of the 211 men and in 55 of the 199 women with mean ages of onset of 43.1 and 46.5 years, respectively. CAD was significantly more common in men (49% v 28%, p < 0.001) and in patients who had smoked cigarettes versus patients who had never smoked (51% v 28%, p < 0.001). After adjusting for age, sex, and current smoking status, there were no significant differences between patients with or without CAD in lipoprotein(a), homocysteine, fibrinogen, plasminogen activator inhibitor-1, white blood cell count, body mass index, glucose, triglyceride or total cholesterol. However, high density lipoprotein (HDL) cholesterol concentrations were significantly lower in those with CAD (6%, 95% confidence interval (CI) 1% to 11%, p = 0.03) and this difference was greater in women than men (12% v 2%, p = 0.041). CONCLUSIONS: These results indicate that emerging coronary risk factors appear not to be associated with CAD in adults with treated familial hypercholesterolaemia, but the strong association with smoking suggests that patients should be identified early in childhood and discouraged from ever starting to smoke. PMID- 15547023 TI - Giant and fusiform aneurysms of coronary arteries following early and adequate treatment of suspected Kawasaki disease. PMID- 15547024 TI - Up regulation of C3, C4, and soluble intercellular adhesion molecule-1 co expresses with high sensitivity C reactive protein in familial hypoalphalipoproteinaemia: further evidence of inflammatory activation. AB - OBJECTIVE: To test the working hypothesis that inflammation underlying precocious and severe coronary atherosclerotic disease in familial hypoalphalipoproteinaemia (FH) can be mediated by up regulation of the innate immune response. METHODS AND RESULTS: 52 patients with FH were compared with 52 healthy controls with regard to immune system markers such as C reactive protein (CRP), soluble intercellular adhesion molecule-1 (sICAM-1), C3c, and C4. Patients differed from controls in their significantly lower concentrations of high density lipoprotein cholesterol (30.2 (4.0) v 50.5 (13.6) mg/dl, p < 0.0001) and apolipoprotein A I (113.2 (19.9) v 148.7 (25.1) mg/dl, p < 0.0001) and their higher triglyceride (139.3 (63.2) v 81.4 (41.7) mg/dl, p < 0.0001) and CRP plasma concentrations (median 0.33 mg/dl, range 0.02-4.66 mg/dl v median 0.07 mg/dl, range 0.02-0.85 mg/dl, p < 0.0001), but not in their total cholesterol and low density lipoprotein cholesterol concentrations. Concentrations of protein complement were higher in patients (C3: 150.8 (42.3) v 101.9 (17.4) mg/dl, p < 0.0001; C4: 35.5 (13.6) v 22.8 (6.4) mg/dl, p < 0.0001) and sICAM-1 concentrations were more than double those found in the controls (335.1 (107.5) v 159.5 (78.2) mg/dl, p < 0.0001). CONCLUSIONS: Increased concentrations of sICAM-1, C3c, and C4 co-express with high concentrations of CRP in FH. The lack of signs and symptoms of inflammation in these patients may suggest that the immune response is up regulated as part of the pro-inflammatory mechanisms that are activated in this atherogenic condition. PMID- 15547025 TI - The safety and effectiveness of a nurse led cardioversion service under sedation. AB - OBJECTIVE: To assess the safety and effectiveness of nurse led elective cardioversion of atrial fibrillation under sedation. DESIGN: Prospective, longitudinal study. SETTING: Cardiac catheterisation laboratory and recovery area of a district general hospital. PATIENTS: 300 patients referred for elective cardioversion of persistent atrial fibrillation. INTERVENTIONS: Pre-procedure evaluations (history, physical examination, blood tests), consent, sedation administration, cardioversions, and post-procedure monitoring until discharge by advanced life support certified coronary care unit nurses trained in the techniques. A doctor was immediately available if required but not present. MAIN OUTCOME MEASURES: Success rates at discharge and at six weeks, energy delivered, number of shocks, dose of sedation, immediate, 24, and 48 hour patient perceptions, complications, waiting times, and cost effectiveness. RESULTS: Cardioversion success rate was 87% at discharge and 48% at six weeks. Mean (SD) cumulative energy was 497 (282) J and number of shocks 1.6 (0.8). Mean (SD) dose of sedation was 23 (9) mg intravenous diazepam. No patient required reversal of sedation, airway support, or medical intervention. Ninety eight per cent of patients had no pain or recall of the procedure. Four patients who were adequately anticoagulated experienced embolic phenomena. Ninety eight per cent of patients would repeat the procedure if necessary. Without requirement for a physician or anaesthetist, waiting times for elective cardioversion fell from three months to under four weeks. There was a significant reduction in the estimated cost of the procedure from 337 pounds sterling with general anaesthesia to 130 pounds sterling with nurse led sedation and cardioversion (p < 0.001). CONCLUSION: With appropriate training, a nurse led cardioversion service with sedation is safe, effective, well tolerated, and cost efficient. PMID- 15547026 TI - Simplified cardioversion service with intravenous midazolam. AB - OBJECTIVE: To assess the results of a new cardioversion service that used intravenous midazolam sedation with cardioversion performed with a biphasic defibrillator by an experienced doctor with a prior review of patients in a pre cardioversion clinic. METHODS: 368 consecutive patients who were treated under the new service over its first 12 months (group 1) were compared with 210 consecutive patients who attended under the old system during the preceding 12 months (group 2). Patients of group 2 had cardioversion under general anaesthesia by junior doctors with a monophasic defibrillator. RESULTS: There were no anaesthetic or respiratory complications in group 1. Of the patients in group 1, 10.3% remembered the shocks, with only 3.5% considering them unpleasant. Cardioversion was successful in 94.6% of group 1 patients after a mean energy of 117 J compared with 81.4% (p < 0.0001) and a mean energy of 242 J (p < 0.0001) for group 2 patients. Cancellations on the day of the procedure were reduced from 24% in group 2 to 3.4% in group 1. CONCLUSIONS: The new service was found to be safe and more efficient. It has led to a large reduction in the waiting time for cardioversion. PMID- 15547027 TI - Relation between coronary pressure derived collateral flow, myocardial perfusion grade, and outcome in left ventricular function after rescue percutaneous coronary intervention. AB - OBJECTIVE: To evaluate the relation between pressure derived coronary collateral flow (PDCF) index and angiographic TIMI (thrombolysis in myocardial infarction) myocardial perfusion (TMP) grade, angiographic collateral grade, and subsequent recovery of left ventricular function after rescue percutaneous coronary intervention (PCI) for failed reperfusion in acute myocardial infarction. METHODS: The pressure wire was used as the guidewire in 38 consecutive patients who underwent rescue PCI between December 2000 and March 2002. Follow up angiography was performed at six months. Baseline and follow up single plane ventriculograms were analysed off line by an automated edge detection technique. A linear model was fitted to assess the relation between 0.1 unit increase in PDCF and change in left ventricular regional wall motion. RESULTS: Patients with TMP 0 grade had significantly higher mean (SD) PDCF than patients with TMP 1-3 (0.30 (0.11) v 0.15 (0.07), p < 0.0001, r = -0.5). A similar relation was observed between TMP grade and coronary wedge pressure (mean (SD) 28 (16) mm Hg with TMP 0 v 9 (7) mm Hg with TMP 1-3, p = 0.001, r = -0.4). Higher PDCF was associated with increased left ventricular end diastolic pressures (0.28 (0.14) with end diastolic pressure > 20 mm Hg v 0.22 (0.09) with end diastolic pressure < 20 mm Hg, p = 0.08, r = 0.2). No correlation was observed between PDCF and Rentrops collateral grade (0.26 (0.13) with grade 0 v 0.25 (0.11) with grades 1 3, p = 0.4, r = -0.06). No linear relation existed between changes in PDCF and changes in left ventricular regional wall motion. CONCLUSION: PDCF in the setting of rescue PCI for failed reperfusion after thrombolysis does not predict improvement in left ventricular function. Increased PDCF and coronary wedge pressure in acute myocardial infarction reflect a dysfunctional microcirculation rather than good collateral protection. PMID- 15547028 TI - Findings of intravascular ultrasound during acute stent thrombosis. AB - OBJECTIVE: To evaluate the potential role of intravascular ultrasound (IVUS) in evaluating patients experiencing an episode of acute stent thrombosis. DESIGN AND SETTING: Prospective observational study in a cardiac catheterisation laboratory in a university teaching hospital. PATIENTS AND INTERVENTIONS: IVUS was used to examine 12 patients undergoing coronary interventions for stent thrombosis to gain further mechanistic insights and to guide treatment. IVUS studies were obtained before and after intervention with a motorised pullback device. MAIN OUTCOME MEASURES: Qualitative and volumetric IVUS analyses. RESULTS: Angiographically, 10 patients had occluded vessels and two patients had intraluminal filling defects within the stent. IVUS showed an occlusive thrombus in all patients. Thrombus volume was 90 (77) mm3, which was 51 (21)% of total stent volume. There was evidence of severe stent underexpansion in most patients and no patient fulfilled standard criteria for optimal stent implantation. Stent malapposition was detected in four patients, edge dissections were seen in two patients, and significant inflow-outflow disease was present in 11 patients. During interventions IVUS findings led to the use of higher pressures or larger balloons than those used during initial stenting in 10 patients. In addition, four patients required additional stenting, whereas a thrombectomy device alone was selected for one patient. After the procedure final minimum stent area (7.1 (2.1) v 5.3 (2) mm2, p < 0.005) and stent expansion (83.2 (17) v 62.1 (15)%, p < 0.005) improved compared with pre-interventional values. However, residual lining thrombus was still visualised in eight patients (25 (19) mm3, accounting for a 17% of final stent volume). CONCLUSIONS: IVUS provides an attractive technique to characterise fully the pattern of stent thrombosis, to identify readily the underlying mechanical predisposing factors, and to guide repeated coronary interventions. PMID- 15547029 TI - Coronary revascularisation outcome questionnaire (CROQ): development and validation of a new, patient based measure of outcome in coronary bypass surgery and angioplasty. AB - OBJECTIVE: To describe the development and scientific validation of a new patient based measure, the coronary revascularisation outcome questionnaire (CROQ), to evaluate health outcomes and quality of life before and after coronary artery bypass grafting and percutaneous transluminal coronary angioplasty. DESIGN AND SETTING: Psychometric validation study conducted with patients from three hospitals in the UK. PATIENTS: Two independent field tests were conducted by postal survey of 714 patients before and 1329 patients after coronary revascularisation to evaluate the measurement properties of the CROQ. METHODS: Qualitative methods including patient interviews were used to develop questionnaire content. A full psychometric evaluation was performed on the survey data. RESULTS: Psychometric tests with the application of stringent criteria confirmed the acceptability (low missing data, good response rates), scaling assumptions (good item convergent and discriminant validity), reliability (good internal consistency and reproducibility), validity (good content and construct validity), and responsiveness of the CROQ. CONCLUSIONS: The CROQ is a practical and scientifically sound patient based measure of outcome developed using psychometric methods. It captures aspects of recovery not addressed in other cardiac questionnaires and has been shown to be a highly responsive instrument that will be useful in evaluating outcomes in clinical trials. PMID- 15547030 TI - Transcatheter closure of persistent ductus arteriosus with the Amplatzer duct occluder in very young symptomatic children. AB - OBJECTIVES: To analyse safety, efficacy, and follow up results of percutaneous closure of persistent ductus arteriosus (PDA) in very young symptomatic children. PATIENTS AND DESIGN: Between March 2000 and March 2003, of 197 patients treated at the authors' institution 18 were symptomatic children aged < or = 3 years old. Seven of these children were < or = 1 year old. Indications for closure were failure to thrive (12 patients) and frequent respiratory infections (six patients). The procedure was carried out under heavy sedation with fluoroscopic control. The Amplatzer duct occluder device was used. Basal physical examinations and echocardiograms were performed before the procedure and at follow up (three, six, and 12 months and yearly thereafter). RESULTS: Mean (SD) age was 18.3 (10) months and mean (SD) weight at closure was 9.1 (2.2) kg. Neither death nor any major complications occurred. Complications occurred in three patients aged < or = 1 year. Two patients had a mild inguinal haematoma. One patient had femoral artery thrombosis that was successfully treated by intravenous urokinase. The mean (SD) follow up was 12.8 (8.5) months. No problems occurred. Patients with recurrent respiratory infections had no significant recurrences and children who had failed to thrive had significantly increased growth. CONCLUSIONS: In experienced hands, percutaneous closure of moderate to large PDA in very young symptomatic children is safe, effectively closes the PDA, and solves clinical problems. PMID- 15547031 TI - Severe aortic insufficiency in a patient with sinus of Valsalva aneurysm invading the interventricular septum. PMID- 15547032 TI - Non-invasive evaluation of atherosclerosis with contrast enhanced 16 slice spiral computed tomography: results of ex vivo investigations. AB - OBJECTIVE: To evaluate the diagnostic accuracy of 16 slice computed tomography (CT) in determining plaque morphology and composition in an experimental setting. The results were compared with histopathological analysis as the reference standard. METHODS: Nine human popliteal arteries derived from amputations because of atherosclerotic disease were investigated with multislice spiral CT (MSCT). Atherosclerotic lesions were morphologically classified (completely or partially occlusive, concentric, eccentric), and tissue densities were determined within these plaques. In addition, vessel dimensions were quantitatively measured. RESULTS: The results were compared with histological analysis. The concordance index kappa for morphological classification was 0.88. Plaque density (n = 51 lesions) was significantly different (p < 0.0001) between lipid rich, fibrotic, and calcified lesions (Stary stage III: n = 2, 58 (8) Hounsfield units (HU); Stary V: n = 11, 50 (21) HU; Stary VI: n = 14, 96 (42) HU; Stary VII: n = 6, 858 (263) HU; Stary VIII: n = 18, 126 (99) HU). The concordance index kappa for the classification of plaques based on density was 0.51. Vessel dimensions had a good correlation (r = 0.98). CONCLUSIONS: 16 slice CT was found to be a reliable non invasive imaging technique for assessing atherosclerotic plaque morphology and composition. Although calcified lesions can be differentiated from non-calcified lesions, the diagnostic accuracy in further subclassifying non-calcified plaques as lipid rich and fibrotic is low, even under experimental conditions. PMID- 15547033 TI - Thrombolysis in patients with a brain tumour. PMID- 15547034 TI - Electron beam computed tomography for the detection of left atrial thrombi in patients with atrial fibrillation. PMID- 15547035 TI - A novel method of assessing Starr-Edwards aortic valve stenosis. PMID- 15547036 TI - Multislice cardiac computed tomography of symmetry bypass connector. PMID- 15547037 TI - Impact of glycaemic and lipid control on outcome after percutaneous coronary interventions in diabetic patients. PMID- 15547038 TI - Late thrombotic occlusion of paclitaxel eluting stent more than one year after stent implantation. PMID- 15547039 TI - Augmented metabolic control improves myocardial diastolic function and perfusion in patients with non-insulin dependent diabetes. PMID- 15547040 TI - Green tea reverses endothelial dysfunction in healthy smokers. PMID- 15547041 TI - Mutations in the long QT gene, KCNQ1, are an uncommon cause of atrial fibrillation. PMID- 15547042 TI - What is the incidence of myocardial necrosis in elective patients discharged on the same day following percutaneous coronary intervention? PMID- 15547043 TI - Vascular brachytherapy versus sirolimus eluting stents for the treatment of in stent restenosis: a prospective registry. PMID- 15547044 TI - Anomalous origin of the right coronary artery from the left anterior descending coronary artery. PMID- 15547045 TI - Theory and practice of defibrillation: (1) Atrial fibrillation and DC conversion. PMID- 15547046 TI - Management of hypertension before, during, and after pregnancy. PMID- 15547047 TI - Closing down: transcatheter closure of intracardiac defects and vessel embolisations. PMID- 15547048 TI - Sertraline is metabolized by multiple cytochrome P450 enzymes, monoamine oxidases, and glucuronyl transferases in human: an in vitro study. AB - The oxidative and conjugative metabolism of sertraline was examined in vitro to identify the enzymes involved in the generation of N-desmethyl, deaminated, and N carbamoyl-glucuronidated metabolites in humans. In human liver microsomes, sertraline was N-demethylated and deaminated by cytochrome P450 (P450) enzymes with overall K(m) values of 98 and 114 microM, respectively, but the intrinsic clearance for N-demethylation was approximately 20-fold greater than for deamination. Using P450 isoform-selective inhibitors and recombinant heterologously expressed enzymes, it was demonstrated that several P450 enzymes catalyzed sertraline N-demethylation, with CYP2B6 contributing the greatest extent, and lesser contributions from CYP2C19, CYP2C9, CYP3A4, and CYP2D6. For deamination, data supported a role for CYP3A4 and CYP2C19. Purified human monoamine oxidases A and B also catalyzed sertraline deamination with comparable K(m) values (230-270 microM). Monoamine oxidase B catalyzed the reaction approximately 3-fold faster than did monoamine oxidase A. Sertraline N-carbamoyl glucuronidation was measured in human liver microsomes in bicarbonate buffer and under a CO2 atmosphere (K(m) = 50 microM) and was catalyzed at the fastest rate by recombinant human UGT2B7. The observation that multiple enzymes appear to be involved in sertraline metabolism suggests that there should be no single agent that could substantially alter the pharmacokinetics of sertraline, nor should there be any single drug-metabolizing enzyme genetic polymorphism (e.g., CYP2D6, CYP2C19, CYP2C9, UGT1A1) that could profoundly impact the pharmacokinetics of sertraline. PMID- 15547049 TI - Saturable distribution of tacrine into the striatal extracellular fluid of the rat: evidence of involvement of multiple organic cation transporters in the transport. AB - The kinetics and mechanism by which tacrine is distributed in the rat brain were examined. Tacrine levels in plasma and striatal extracellular fluid were used to evaluate the pharmacokinetics of this process. The K(D,brain) was decreased with the dose for tacrine, indicating that the distribution to the brain is saturable. The uptake of organic cations such as choline, 1-methyl-4-phenylpyridinium (MPP), tetraethylammonium (TEA), and carnitine was inhibited by the addition of tacrine to cultures of mouse immortalized brain capillary endothelial cells. In addition, the apical to basal transport and basal to apical transport of tacrine were inhibited by the addition of organic cations to cultures of LLC-PK1 cells, suggesting that tacrine transport across the blood-brain barrier (BBB) is mediated by organic cation transport system(s). Consistent with the in vitro results, a standard reverse transcription-polymerase chain reaction procedure was able to amplify the message of mOCT2 and mOCTN2, but not mOCT1, in MBEC4 (mouse brain microvessel endothelial cell line 4) cells. Similarly, mRNAs for rOCT2 and rOCTN2 were present in representative rat brain samples. To determine whether OCT2 and/or OCTN2 transport tacrine, these transporters were cloned and then transfected in SK-HEP1 and HEK 293 cells. The uptake of choline, MPP, and TEA was inhibited by the presence of tacrine in rOCT2-expressing SK-HEP1 cells, whereas the uptake of carnitine was inhibited by the presence of tacrine in rOCTN2 expressing HEK 293 cells. Collectively, these observations suggest that the transport of tacrine across the BBB is mediated, at least in part, by multiple organic cation transport systems in rats. PMID- 15547050 TI - Absorption and elimination of formate following oral administration of calcium formate in female human subjects. AB - Calcium formate is a water-soluble salt of an essential mineral nutrient with potential for use as a dietary calcium supplement. Formate ion is a product of endogenous and xenobiotic metabolism, but sustained high plasma formate concentrations (such as occur in cases of methanol poisoning) are toxic to the retina and optic nerve. Humans and primates have reduced capacity for formate oxidation compared with rodents and dogs and are thus more sensitive to methanol (and formate) intoxication. To assess the potential for accumulation of formate ion upon repeated administration of calcium formate as a potential dietary calcium supplement, we measured plasma concentrations of formate in 14 adult human subjects before and after oral administration of a single large dose of calcium formate (3900 mg; ca. 3-6 times the anticipated dose for calcium supplementation). Plasma formate concentrations increased briskly from 0.024 +/- 0.008 mM (endogenous formate) to reach C(max) (0.50 +/- 0.04 mM) at 60 min postdose and then declined with a half-life of 59 +/- 7 min. By 225 min postdose, plasma formate concentration had returned to baseline. With such a short half life, repeated use of calcium formate as a dietary supplement, even three times daily, should not lead to progressive accumulation of formate. These findings are discussed in light of the production of formate by endogenous and xenobiotic metabolism and the kinetics of formate during methanol poisoning. PMID- 15547052 TI - A tentative step towards healthy public policy. PMID- 15547051 TI - Extrahepatic metabolism of carbamate and organophosphate thioether compounds by the flavin-containing monooxygenase and cytochrome P450 systems. AB - The cytochrome P450 (P450) and flavin-containing monooxygenase (FMO) enzymes are the major oxidative enzymes in phase I metabolism. Many organophosphate and carbamate thioether compounds are excellent substrates for these enzymes. Stereoselective sulfoxidation of fenthion and methiocarb by human liver, kidney, and microsomes was investigated. A high level of stereoselectivity in the formation of fenthion +-sulfoxide was observed in kidney and intestinal microsomes. This activity was not inhibited by the P450 inhibitor 1 aminobenzotriazole but was dramatically reduced following mild heat treatment. In liver, fenthion was metabolized to its sulfoxide in a nonstereoselective manner, and the activity was sensitive to both 1-aminobenzotriazole and heat treatment. The carbamate pesticide methiocarb also was sulfoxidated with a high degree of stereoselectivity in human kidney microsomes. Human liver microsomes formed both stereoisomers in equal amounts. Sulfoxide formation in kidney was not inhibited by 1-aminobenzotriazole but was abolished in liver microsomes. Formation of methiocarb sulfoxides was not observed in intestinal microsomes. The relative contribution of FMO1 and FMO3 to the sulfoxidation of carbophenothion, demeton-O, ethiofencarb, fonofos, and methiocarb also was investigated by using baculovirus expressed recombinant proteins. FMO1 showed the highest catalytic activity for all pesticides. This study indicates that FMO1 may have a bigger role in extrahepatic metabolism than previously thought. PMID- 15547053 TI - Assessing psychosocial/quality of life outcomes in screening: how do we do it better? PMID- 15547054 TI - A glossary of terms for navigating the field of social network analysis. AB - Social network analysis is the study of social structure. This glossary introduces basic concepts in social network analysis. It is designed to help researchers to be more discriminating in their thinking and choice of methods. PMID- 15547055 TI - Relation between increased numbers of safe playing areas and decreased vehicle related child mortality rates in Japan from 1970 to 1985: a trend analysis. AB - OBJECTIVES: To examine vehicle related mortality trends of children in Japan; and to investigate how environmental modifications such as the installation of public parks and pavements are associated with these trends. DESIGN: Poisson regression was used for trend analysis, and multiple regression modelling was used to investigate the associations between trends in environmental modifications and trends in motor vehicle related child mortality rates. SETTING: Mortality data of Japan from 1970 to 1994, defined as E-code 810-23 from 1970 to 1978 and E810-25 from 1979 to 1994, were obtained from vital statistics. Multiple regression modelling was confined to the 1970-1985 data. Data concerning public parks and other facilities were obtained from the Ministry of Land, Infrastructure, and Transport. SUBJECTS: Children aged 0-14 years old were examined in this study and divided into two groups: 0-4 and 5-14 years. MAIN RESULTS: An increased number of public parks was associated with decreased vehicle related mortality rates among children aged 0-4 years, but not among children aged 5-14. In contrast, there was no association between trends in pavements and mortality rates. CONCLUSIONS: An increased number of public parks might reduce vehicle related preschooler deaths, in particular those involving pedestrians. Safe play areas in residential areas might reduce the risk of vehicle related child death by lessening the journey both to and from such areas as well as reducing the number of children playing on the street. However, such measures might not be effective in reducing the vehicle related mortalities of school age children who have an expanded range of activities and walk longer distances. PMID- 15547056 TI - Post-disaster health effects: strategies for investigation and data collection. Experiences from the Enschede firework disaster. AB - BACKGROUND: Public health policy is increasingly concerned with the care for victims of a disaster. This article describes the design and implementation of an epidemiological study, which seeks to match care services to the specific problems of persons affected by a large scale incident. The study was prompted by the explosion of a firework depot in Enschede, the Netherlands. STUDY POPULATION: All those directly affected by this incident (residents, emergency services personnel, and people who happened to be in the area at the time), some of whom suffered personal loss or injury. The project investigates both the physical and psychological effects of the disaster, as well as the target group's subsequent call on healthcare services. STUDY DESIGN: A questionnaire based follow up survey of those directly affected and an ongoing monitoring of health problems relying on reports from healthcare professionals. The follow up survey started three weeks after the incident and was repeated 18 months and almost four years after the incident. The monitoring is conducted over a four year period by general practitioners, the local mental health services department, occupational health services, and the youth healthcare services department. It provides ongoing information. RESULTS AND CONCLUSIONS: The results of the study are regularly discussed with healthcare professionals and policy makers, and are made known to the research participants. The paper also explains the considerations that were made in designing the study to help others making up their research plans when confronted with possible health effects of a disaster. PMID- 15547057 TI - SARS related preventive and risk behaviours practised by Hong Kong-mainland China cross border travellers during the outbreak of the SARS epidemic in Hong Kong. AB - OBJECTIVES: To investigate patterns of behaviours and attitudes related to SARS prevention in the Hong Kong cross border traveller population. SETTINGS: A survey was carried out at the Hong Kong-China cross border checkpoint in the middle of the epidemic. PARTICIPANTS: A total of 839 Hong Kong adult residents returning to Hong Kong from mainland China were surveyed. MAIN OUTCOME MEASURES: Practice of preventive measures and relevant behaviours and attitudes. RESULTS: Around 40% of the respondents were using masks all or most of the time in public places or washing their hands frequently (>10 times per day) and about one third avoided visiting crowded places in mainland China. Such figures were however lower than those practised by the general public in Hong Kong. SARS related perceptions, such as perceived risk of transmission and efficacy, etc, were associated with mask use and not visiting crowded places, but not with hand washing, which was associated with duration of stay. Gender differences were also observed. Around 70% of the travellers would have delayed medical consultation for influenza-like illness in China; 12.7% would not wear masks during such episodes of illness. Furthermore, about 30% of the respondents used to wear masks in Hong Kong but not in mainland China. CONCLUSIONS: The findings have implications on cross border prevention of SARS. It seems that those travelling during the SARS epidemic were a "self selected" group, and they were using less preventive measures. Special attention and intervention need to be provided to travellers to prevent a second wave cross border transmission of the disease. PMID- 15547059 TI - The Gatehouse Project: can a multilevel school intervention affect emotional wellbeing and health risk behaviours? AB - STUDY OBJECTIVE: The aim of this study was to determine the effect of a multilevel school based intervention on adolescents' emotional wellbeing and health risk behaviours. DESIGN: School based cluster randomised controlled trial. Students were surveyed using laptop computers, twice in the first year of intervention and annually thereafter for a further two years. SETTING: Secondary schools. PARTICIPANTS: 2678 year 8 students (74%) participated in the first wave of data collection. Attrition across the waves was less than 3%, 8%, and 10% respectively with no differential response rate between intervention and control groups at the subsequent waves (98% v 96%; 92% v 92%, and 90% v 89% respectively). MAIN RESULTS: A comparatively consistent 3% to 5% risk difference was found between intervention and control students for any drinking, any and regular smoking, and friends' alcohol and tobacco use across the three waves of follow up. The largest effect was a reduction in the reporting of regular smoking by those in the intervention group (OR 0.57, 0.62, and 0.72 at waves 2, 3, and 4 respectively). There was no significant effect of the intervention on depressive symptoms, and social and school relationships. CONCLUSIONS: While further research is required to determine fully the processes of change, this study shows that a focus on general cognitive skills and positive changes to the social environment of the school can have a substantial impact on important health risk behaviours. PMID- 15547060 TI - Cost effectiveness of a community based exercise programme in over 65 year olds: cluster randomised trial. AB - OBJECTIVE: To assess the cost effectiveness of a community based exercise programme as a population wide public health intervention for older adults. DESIGN: Pragmatic, cluster randomised community intervention trial. SETTING: 12 general practices in Sheffield; four randomly selected as intervention populations, and eight as control populations. PARTICIPANTS: All those aged 65 and over in the least active four fifths of the population responding to a baseline survey. There were 2283 eligible participants from intervention practices and 4137 from control practices. INTERVENTION: Eligible subjects were invited to free locally held exercise classes, made available for two years. MAIN OUTCOME MEASURES: All cause and exercise related cause specific mortality and hospital service use at two years, and health status assessed at baseline, one, and two years using the SF-36. A cost utility analysis was also undertaken. RESULTS: Twenty six per cent of the eligible intervention practice population attended one or more exercise sessions. There were no significant differences in mortality rates, survival times, or admissions. After adjusting for baseline characteristics, patients in intervention practices had a lower decline in health status, although this reached significance only for the energy dimension and two composite scores (p<0.05). The incremental average QALY gain of 0.011 per person in the intervention population resulted in an incremental cost per QALY ratio of 17 174 (95% CI = 8300 to 87 120). CONCLUSIONS: Despite a low level of adherence to the exercise programme, there were significant gains in health related quality of life. The programme was more cost effective than many existing medical interventions, and would be practical for primary care commissioning agencies to implement. PMID- 15547061 TI - Health proverbs. PMID- 15547062 TI - Increase of regional total cancer incidence in north Sweden due to the Chernobyl accident? AB - STUDY OBJECTIVE: Is there any epidemiologically visible influence on the cancer incidence after the Chernobyl fallout in Sweden? DESIGN: A cohort study was focused on the fallout of caesium-137 in relation to cancer incidence 1988-1996. SETTING: In northern Sweden, affected by the Chernobyl accident in 1986, 450 parishes were categorised by caesium-137 deposition: < 3 (reference), 3-29, 30 39, 40-59, 60-79, and 80-120 kiloBecquerel/m(2). PARTICIPANTS: All people 0-60 years living in these parishes in 1986 to 1987 were identified and enrolled in a cohort of 1 143 182 persons. In the follow up 22 409 incident cancer cases were retrieved in 1988-1996. A further analysis focused on the secular trend. MAIN RESULTS: Taking age and population density as confounding factors, and lung cancer incidence in 1988-1996 and total cancer incidence in 1986-1987 by municipality as proxy confounders for smoking and time trends, respectively, the adjusted relative risks for the deposition categories were 1.00 (reference < 3 kiloBecquerel/m(2)), 1.05, 1.03, 1.08, 1.10, and 1.21. The excess relative risk was 0.11 per 100 kiloBecquerel/m(2) (95% CI 0.03 to 0.20). Considering the secular trend, directly age standardised cancer incidence rate differences per 100 000 person years between 1988 to 1996 and the reference period 1986-1987, were 30.3 (indicating a time trend in the reference category), 36.8, 42.0, 45.8, 50.1, and 56.4. No clear excess occurred for leukaemia or thyroid cancer. CONCLUSIONS: Unless attributable to chance or remaining uncontrolled confounding, a slight exposure related increase in total cancer incidence has occurred in northern Sweden after the Chernobyl accident. PMID- 15547063 TI - Fear of racism and health. PMID- 15547064 TI - Is the effect of work stress on cardiovascular mortality confounded by socioeconomic factors in the Valmet study? PMID- 15547065 TI - Possible association between heavy computer users and glaucomatous visual field abnormalities: a cross sectional study in Japanese workers. AB - STUDY OBJECTIVE: To study the association between computer use and visual field abnormalities (VFA) and to assess whether heavy computer users have an increased risk of glaucoma. DESIGN: Cross sectional multicentre study. Subjects and observation procedures: A total of 10 202 randomly selected Japanese workers (mean (SD) age 43.2 (9.8) years) were screened for VFA using the frequency doubling technology perimetry (FDT-VFA), in addition to undergoing a general medical check up, and then ophthalmologically examined. Information about their computer use and refractive errors was obtained from a questionnaire and interview, respectively. MAIN RESULTS: As a result of FDT test, 522 and 8602 subjects were positive and negative for FDT-VFA, respectively. A significant (p = 0.004) interaction was found between computer use and refractive errors regarding the risk of FDT-VFA. In stratified analysis, heavy computer users with refractive errors showed a significant positive association with FDT-VFA (odds ratio (OR) = 1.74, 95% confidence interval (CI) 1.28 to 2.37), while those without refractive errors did not. Comparison of 165 subjects with an ophthalmological diagnosis of glaucoma and 2918 controls showed that the OR for glaucoma of heavy computer users with refractive errors was 1.82 (95% CI 1.06 to 3.12). Of 165 subjects with glaucoma, 141 had refractive errors, especially myopia (96.4%, 136 of 141). CONCLUSIONS: Although there are limitations to this study, such as its cross sectional design, heavy computer users with refractive errors seem to have an increased risk of FDT-VFA. Glaucoma might be involved in an underlying disease and myopia in a risk factor for FDT-VFA. PMID- 15547066 TI - Life course measures of socioeconomic position and self reported health at age 50: prospective cohort study. PMID- 15547067 TI - Confounding by socioeconomic position remains after adjusting for neighbourhood deprivation: an example using smoking and mortality. PMID- 15547068 TI - Multiple sclerosis after infectious mononucleosis: record linkage study. AB - OBJECTIVE: To ascertain if infectious mononucleosis is a risk factor for the development of multiple sclerosis (MS); and, if it is, whether its effect is close to or remote in time from the onset of MS. DESIGN: Analysis of database of linked abstracts of records of hospital admission and death. SETTING: Health region in central southern England. MAIN OUTCOME MEASURE: Ratio of rate of MS in a cohort of people admitted to hospital with infectious mononucleosis to the rate in a comparison cohort. RESULTS: Considering all time intervals from admission with infection to admission with MS, there was a non-significant increase of risk of MS in the infectious mononucleosis cohort (rate ratio 2.17, 95% confidence intervals 0.79 to 4.77). At the interval of 10 years or more, there was a significant increase in risk of MS (rate ratio 4.01, 1.48 to 8.93). The mean time from infectious mononucleosis to first admission with MS was 14 years. CONCLUSION: This study adds support to the evidence that Epstein-Barr virus, the cause of infectious mononucleosis, is associated with MS. Its role is probably as an initiator of the disease process of MS, or as a contributor to its early development, rather than as an activator of latent, existing disease. PMID- 15547069 TI - Prevalence of comorbid psychiatric illness and substance misuse in primary care in England and Wales. AB - STUDY OBJECTIVE: To estimate the annual period prevalence of co-occurring psychiatric illness and substance misuse among patients in primary care. DESIGN: Analysis of the general practice research database. SETTING: England and Wales, 1993-1998. PARTICIPANTS: Registered patients at 230 general practices representing 3.1% of the population. A comorbid case was defined as one with both a psychiatric diagnosis and substance misuse diagnosis (not including alcohol or tobacco) within a calendar year. A potentially chronic comorbid case was one that met this definition and, in addition, was treated in subsequent years for either a psychiatric condition or substance misuse. MAIN RESULTS: The annual period prevalence of comorbidity increased from 50/100 000 patient years of exposure (PYE) to 80/100 000 PYE, an increase of 62% during the study period. Rates of comorbid psychoses, comorbid schizophrenia, and comorbid paranoia increased by 147%, 128%, and 144%. The average age of comorbid cases decreased from 38 years to 34 years. Over 80% of comorbid cases were newly diagnosed in each study year, although many are treated in subsequent years for either psychiatric illness or substance misuse. CONCLUSIONS: This study provides data on the nature and extent of comorbidity in primary care in England and Wales. As the comorbidity rate is increasing by about 10% each year, and as comorbid cases are becoming younger, it is probable that the comorbidity rate will have increased beyond the study end point. PMID- 15547070 TI - Poorer self assessed health in a prospective study of men with screen detected abdominal aortic aneurysm: a predictor or a consequence of screening outcome? AB - STUDY OBJECTIVES: To assess the extent to which poorer self assessed health in men in whom an abdominal aortic aneurysm (AAA) is detected at screening is a consequence or a predictor of screening outcome. DESIGN: Prospective study. SETTING: Community based screening. PARTICIPANTS: 23 654 men who attended for AAA screening as part of the UK multicentre aneurysm screening study completed a measure of self assessed health before screening. A total of 1156 had an aneurysm detected. A sub-sample of screened men (571 with an aneurysm and 609 with a normal aorta) also completed the measure of self assessed health six weeks after screening. MAIN RESULTS: Men in whom an aneurysm was detected at screening perceived their health to be poorer before screening than those with a normal aorta. Adjusting for risk factors for AAA made no difference to this RESULT: self assessed health remained a strong predictor of having an aneurysm (odds ratio 1.7 comparing the extreme quartiles of self assessed health, 95% confidence intervals: 1.4 to 2.0). Men with an aneurysm also perceived their health to be poorer after screening had detected their aneurysms, but only to an extent in line with their pre-screening perceptions. CONCLUSIONS: Self assessed health seems to predict having an aortic aneurysm, independently of known risk factors. This emphasises the importance of assessing baseline perceptions of health to prevent erroneously inferring that poorer self assessed health in those who screen positive is a consequence as compared with a predictor of screening outcome. PMID- 15547071 TI - Lipid lowering drugs prescription and the risk of peripheral neuropathy: an exploratory case-control study using automated databases. AB - STUDY OBJECTIVE: Although lipid lowering drugs are effective in preventing morbidity and mortality from cardiovascular events, the extent of their adverse effects is not clear. This study explored the association between prescription of lipid lowering drugs and the risk of peripheral neuropathy. DESIGN: A population based case-control study was carried out by linkage of several automated databases. SETTING: Resident population of a northern Italian Province aged 40 years or more. PARTICIPANTS: Cases were patients discharged for peripheral neuropathy in 1998-1999. For each case up to 20 controls were randomly selected among those eligible. Altogether 2040 case patients and 36 041 controls were included in the study. Exposure ascertainment: Prescription drug database was used to assess exposure to lipid lowering drugs at any time in the one year period preceding the index date. ANALYSIS: Conditional logistic regression model for matched data was used to estimate the risk of peripheral neuropathy associated with exposure to statins, fibrates, and other lipid lowering drugs. MAIN RESULTS: Weak but significant effects of lipid lowering drugs as a whole (matched odds ratio: 1.27; 95% confidence intervals: 1.05 to 1.55), statins (1.19; 1.00 to 1.40), and fibrates (1.49; 1.03 to 2.17) were observed. Significant linear trends towards increased risk at increased exposure to both statins and fibrates were observed. CONCLUSIONS: The use of both statins and fibrates was associated with the risk of peripheral neuropathy. The primary purpose of this exploratory study is signal generation. This requires further investigations to evaluate the causal role of lipid lowering drugs on the onset of peripheral neuropathy. PMID- 15547072 TI - Are there socioeconomic gradients in the quality of data held by UK cancer registries? PMID- 15547073 TI - Qualia years (qy) [corrected]--not years--should be the unit of measurement of QALYs, DALYs, life expectancy, and life. PMID- 15547074 TI - Relation of rainfall pattern and epidemic leptospirosis in the Indian state of Kerala. PMID- 15547075 TI - Do statins offer therapeutic potential in inflammatory arthritis? PMID- 15547076 TI - Systemic juvenile idiopathic arthritis presenting in a young child with long term disability as an adolescent. PMID- 15547077 TI - Adipose tissue at entheses: the rheumatological implications of its distribution. A potential site of pain and stress dissipation? AB - OBJECTIVES: To describe the distribution of adipose tissue within and adjacent to entheses in order to assess its functional significance at attachment sites. METHODS: Entheses were removed from 29 different sites in the limbs of formalin fixed, elderly, dissecting room cadavers and the samples prepared for paraffin and/or methylmethacrylate histology. Entheses from four young volunteers with no history of significant musculoskeletal injury were examined by magnetic resonance imaging using T(1) weighted sequences. RESULTS: Adipose tissue was present at several different sites at numerous entheses. Many tendons/ligaments lay on a bed of well vascularised, highly innervated, "insertional angle fat". Endotenon fat was striking between fascicles, where entheses flared out at their attachments. It was also characteristic of the epitenon, where it occurred in conjunction with lamellated and Pacinian corpuscles. Fat filled, meniscoid folds often protruded into joint cavities, immediately adjacent to attachment sites. CONCLUSION: Adipose tissue is a common feature of normal entheses and should not be regarded as a sign of degeneration. It contributes to the increase in surface area of attachment sites, promotes movement between tendon/ligament and bone, and forms part of an enthesis organ that dissipates stress. The presence of numerous nerve endings in fat at attachment sites suggests that it has a mechanosensory role and this could account for the rich innervation of many entheses. Because damage to fat is known to lead to considerable joint pain, our findings may be important for understanding the site of pain in enthesopathies. PMID- 15547078 TI - Increased FcgammaRII expression and aberrant tumour necrosis factor alpha production by mature dendritic cells from patients with active rheumatoid arthritis. AB - OBJECTIVES: To investigate potential differences in phenotype and behaviour of immature (iDC) and mature dendritic cells (mDC) from patients with RA and healthy subjects. METHODS: iDC and mDC were derived from blood monocytes of patients with RA and healthy controls following standardised protocols. FACS was used to analyse expression of FcgammaRI, II, and III and molecules to characterise DC. Discrimination between FcgammaRIIa and FcgammaRIIb was achieved by RT-PCR. Immunohistochemistry was performed on synovial biopsy specimens of three patients with RA and three healthy controls. TNFalpha production by iDC and mDC upon FcgammaR dependent stimulation was compared between patients with RA and controls by ELISA. RESULTS: iDC from patients with active RA but not from patients with inactive RA or healthy controls markedly up regulated FcgammaRII. mDC from patients with active RA also lacked the physiological down regulation of FcgammaRII that occurs upon maturation in both control groups. RT-PCR analysis confirmed the increased expression of FcgammaRII in RA-especially marked for FcgammaRIIb. FcgammaR dependent stimulation of DC using antigen-IgG immune complexes (IC) significantly increased TNFalpha production by DC from healthy subjects, but significantly decreased TNFalpha by DC from patients with RA. Overlapping expression patterns between FcgammaRII and DC-LAMP in the synovial tissue of patients with RA imply that in vivo, also, mature DC express increased levels of FcgammaRIIb. CONCLUSION: The presence and altered characteristics of DC during active RA suggest that DC help to modulate autoimmunity in RA. Further studies should elucidate the role of local factors in altering the function of DC in RA and in increasing expression of FcgammaRII. PMID- 15547079 TI - Divergent roles of nitrergic and prostanoid pathways in chronic joint inflammation. AB - BACKGROUND: Nitrergic and prostanoid pathways have both been implicated in inflammatory processes. OBJECTIVE: To investigate their respective contributions in a rat model of chronic arthritis. METHODS: Male Wistar rats (n = 4-6/group) received either an intra-articular injection of 2% carrageenan/4% kaolin (C/K) or intra- and periarticular injections of Freund's complete adjuvant (FCA; 10 mg/ml M tuberculosis). Joint diameter, urinary nitric oxide metabolites (NO(x)), and prostaglandin E(2) (PGE(2)) levels were measured as indices of the inflammatory process. A prophylactic and therapeutic (day 5) dose ranging study of an inducible nitric oxide synthase inhibitor, L-N-(1-iminoethyl)-lysine (L-NIL), and a cyclo-oxygenase-2 (COX-2) inhibitor, SC-236, was performed with the drugs given subcutaneously. Submaximal doses were identified and used for combination studies. Appropriate vehicle controls were included. RESULTS: L-NIL and SC-236 dose dependently inhibited C/K induced acute joint swelling, the magnitude being greatest when they were given in combination. Both prophylactic and therapeutic administration of SC-236 in the FCA induced model of chronic arthritis produced a dose dependent reduction in all the measures assessed. However, although L-NIL demonstrated similar dose dependent inhibition of urinary NO(x) and PGE(2) levels, joint swelling was significantly exacerbated in this model. Co administration of the inhibitors nullified the benefits of SC-236. CONCLUSION: Whereas COX-2 derived prostaglandins are proinflammatory in both acute and chronic joint inflammation, NO seems to have divergent roles, being anti inflammatory in chronic and proinflammatory in acute joint inflammation. PMID- 15547080 TI - Atorvastatin reduces arterial stiffness in patients with rheumatoid arthritis. AB - BACKGROUND: Chronic systemic inflammation may contribute to accelerated atherosclerosis and increased arterial stiffness in patients with rheumatoid arthritis (RA). In addition to lowering cholesterol, statins have immunomodulatory effects which may be especially beneficial in patients with RA who have systemic immune activation. OBJECTIVE: To investigate the effect of atorvastatin on the augmentation index (AIx: a measure of arterial stiffness) and systemic inflammation in RA. METHODS: 29 patients with RA (mean (SD) age 55 (13) years) with moderately active disease of long duration were studied. AIx, lipid levels, serum inflammatory markers, and disease activity score were measured before and after 12 weeks of atorvastatin 20 mg daily. RESULTS: AIx improved significantly from 34.1 (11.6)% to 29.9 (11)% (p = 0.0002), with the greatest improvements in AIx occurring in those subjects with the highest disease activity scores (r = -0.5, p = 0.007). Total and LDL cholesterol were reduced from 5.5 (0.9) to 3.9 (0.7) mmol/l and 3.3 (0.8) to 1.9 (0.6) mmol/l, respectively (p = 0.0001). Serum inflammatory markers remained unchanged during the study. CONCLUSIONS: Atorvastatin significantly reduced arterial stiffness in patients with RA. The greatest improvements were seen in patients with more active disease, suggesting that, in addition to the beneficial effects of cholesterol reduction, immune modulation may contribute to the cardioprotective effect of statins. PMID- 15547081 TI - Bone mineral density in patients with rheumatoid arthritis: relation between disease severity and low bone mineral density. AB - OBJECTIVE: To examine variables associated with bone mineral density (BMD) in patients with rheumatoid arthritis (RA). METHODS: We investigated 373 patients with low to moderately active RA. Patients with low disease activity were recruited from a cohort of patients in clinical remission. Patients with moderately active disease were included in a trial comparing the effects of long term high intensity exercise programme and conventional physical therapy. Demographic and clinical data were collected. Bone mineral density (BMD) was measured by means of dual x ray absorptiometry (DXA). Associations between demographic and clinical measurements on the one hand and BMD on the other were investigated in regression analyses. RESULTS: The patient group consisted of middle aged, mainly female, patients. The median (interquartile range) disease duration was 7 (4 to 13) years, the mean disease activity score (standard deviation) was 3.2 (1.4). Of the group, 66% was rheumatoid factor positive, and 83% (n = 304) had never used corticosteroids. The median Larsen score of hands and feet was 27 (5 to 61). Greater age and low body mass index were related to low BMD at the hip and spine. High Larsen score for hands and feet was significantly associated with low BMD at the hip. The use of corticosteroids was not independently associated with BMD. The results of the multiple regression analyses also applied to the subgroup of corticosteroid naive patients. CONCLUSION: BMD data of patients with low to moderately active RA demonstrated an association between high radiological RA damage and low BMD at the hip, which suggests an association between the severity of RA and the risk of generalised bone loss, which also occurred in corticosteroid naive patients. PMID- 15547082 TI - HLA-DMA*0103 and HLA-DMB*0104 alleles as novel prognostic factors in rheumatoid arthritis. AB - OBJECTIVE: To evaluate HLA-DM alleles as markers for disease severity in rheumatoid arthritis (RA). METHODS: Two distinct cohorts of patients with RA were oligotyped for HLA-DB1 and HLA-DM genes using PCR amplified genomic DNA with sequence specific oligonucleotide probes. Cohort 1 comprised 199 unselected patients with RA (mean (SD) age 45.5 (13.5) years; disease duration 11.9(8.8) years), whose disease severity was assessed using Larsen score on hand and foot radiographs. Cohort 2 comprised 95 patients with severe RA and 70 patients with benign RA according to the Larsen method. RESULTS: In cohort 1, after stratification according to DRB1 genotypes, patients positive for HLA-DMA*0103 and negative for HLA-DRB1*04 tended to have greater articular damage on hands and wrists (p = 0.07 by Mann-Whitney U test) and reached statistical significance for the Larsen score per year (p = 0.05). This association between HLA-DMA*0103 and articular damage was especially observed in patients with HLA-DRB1*01. Similarly, HLA-DMB*0104 positive patients had higher Larsen score on hands and wrists (p = 0.02). This association was even stronger in DRB1*04 positive patients (p = 0.005). In cohort 2, HLA-DMA*0103 was associated with severe RA in patients negative for HLA-DRB1*04 (OD = 5.4; p = 0.014). HLA-DMB*0104 allele frequency tended to be higher in patients with severe RA but without reaching significance. CONCLUSION: This is the first study evaluating the role of HLA-DM genes in the severity of RA. Our results suggest that HLA-DMA*0103 and HLA-DMB*0104 alleles may represent new genetic markers of RA severity. The HLA-DMA*0103 allele tends to be associated with patients with RA negative for DRB1*04 and could predict a more severe form of disease especially in HLA-DRB1*01 positive patients. The HLA DMB*0104 allele could have an additive effect in HLA-DRB1*04 patients. Combined determination of HLA-DM and HLA-DRB1 alleles could facilitate identification of patients likely to have a poor disease course. PMID- 15547083 TI - Rheumatoid factor and anticitrullinated protein antibodies in rheumatoid arthritis: diagnostic value, associations with radiological progression rate, and extra-articular manifestations. AB - BACKGROUND: Autoantibodies such as rheumatoid factor (RF) and anticitrullinated protein antibodies can be detected in rheumatoid arthritis (RA) sera. OBJECTIVE: To determine the diagnostic values of RF, anticitrullinated protein antibodies, and the shared epitope (SE), and their associations with radiological progression rates and extra-articular manifestations. METHODS: Population 1 consisted of sera from 315 patients, consecutively sent for detection of anticitrullinated protein antibodies, of which 264 were used to determine the sensitivity and specificity of RF and of antibodies against three synthetic citrullinated peptides: peptide A (pepA), peptide B (pepB), and CCP2. Population 2 consisted of sera from 180 longstanding RA patients and was used to determine associations of RA associated antibodies and the SE with radiological progression rates and extra-articular manifestations. Antibodies to pepA and pepB were detected by line immunoassay, and antibodies to CCP2 by ELISA. HLA Class II typing was performed by LiPA. RESULTS: In population 1, we defined adapted cut offs corresponding to a specificity of >/=98.5%. This yielded the following sensitivities: RF 12.8%; anti pepA antibodies 63.6%; anti-pepB antibodies 54.2%; and anti-CCP2 antibodies 73.7%. In population 2, significant differences in radiological progression rates were found between positive and negative patients for different RA antibodies and the SE. RF, but not anticitrullinated protein antibodies or the SE, were more frequent in patients with extra-articular manifestations. CONCLUSION: A valid comparison of RA associated antibodies shows superior sensitivity of the anticitrullinated protein antibodies compared with RF. The presence of RA associated antibodies and the SE are indicative for poorer radiological outcome, and presence of extra-articular manifestations is associated with RF but not with anticitrullinated protein antibodies. PMID- 15547084 TI - Health status of patients with ankylosing spondylitis: a comparison with the general population. AB - OBJECTIVE: To examine the subjective health in patients with ankylosing spondylitis (AS) compared with the general population, and to explore the associations between health status and age, sex of the patients, and educational level in AS. METHODS: Health status was assessed with a generic instrument (SF 36) in 314 patients with AS and in 2323 people from the general population. Subgroup analyses were performed according to age, sex, and educational level. Standard difference scores (s-scores) were calculated to ensure the clinical meaningfulness of the norm based comparisons. RESULTS: Both men and women with AS reported significantly impaired health on all scales of the SF-36. Women reported significantly worse health on physical health domains. However, when calculating differences from the general population, numerically larger s-scores were found for men (except for physical role and vitality). The relative impact of AS seems to diminish with increasing age. In AS, better health was significantly associated with higher education across all scales. Deviations from the general population on the non-physical health aspects were especially pronounced in patients with low education. CONCLUSIONS: All key dimensions of health are affected by AS. The physical aspects seem to be most severely affected, but in the less educated group of patients, the disease impact on the mental health aspects was also considerable. Evaluation and management planning should take the complexity of AS into consideration. The focus on physical function should be maintained, and additional attention should be paid to the mental and social consequences of AS. PMID- 15547085 TI - Chondrocyte number and proteoglycan synthesis in the aging and osteoarthritic human articular cartilage. AB - OBJECTIVE: To correlate the number of chondrocytes in healthy and osteoarthritic human articular cartilage with age, and to evaluate the influence of donor age on total proteoglycan synthesis. METHODS: Chondrocytes were isolated from human articular cartilage derived from hip joints with and without osteoarthritic lesions. The cell number was normalised to cartilage sample wet weight. In addition, the influence of age on chondrocyte numbers was assessed histomorphometrically. Chondrocytes were grown as monolayer cultures for seven days in a chemically defined serum-free basal medium. Total proteoglycan synthesis was measured by [(35)S]sulphate incorporation into newly synthesised macromolecules. RESULTS: Chondrocyte numbers in healthy cartilage decreased significantly with advancing age (r = -0.69, p<0.0001). In contrast to healthy specimens, chondrocyte numbers were decreased in osteoarthritic cartilage irrespective of and unrelated to age, and differed markedly, by an average of 38%, from the cell numbers found in healthy individuals (p<0.0001). Regarding synthesis of matrix macromolecules, no dependence on patients' age, either in healthy or in osteoarthritic specimens, could be observed. CONCLUSIONS: Under the experimental conditions employed, chondrocytes from healthy and osteoarthritic joints synthesised comparable amounts of cartilage macromolecules, independent of age or underlying osteoarthritic disease. Thus the decrease in chondrocyte number in aging and osteoarthritic joints could be a crucial factor in limiting tissue replenishment. PMID- 15547086 TI - Dehydroepiandrosterone suppresses interleukin 10 synthesis in women with systemic lupus erythematosus. AB - OBJECTIVE: To study the effects of dehydroepiandrosterone (prasterone, DHEA) 200 mg/day on cytokine profiles in adult women with active systemic lupus erythematosus (SLE). METHODS: In a double blind, randomised, placebo controlled study conducted as part of a larger multicentre study, 30 adult women with active SLE received oral DHEA 200 mg/day or placebo for 24 weeks. Baseline prednisone (<10 mg/day) and other concomitant SLE medications were to remain constant. The levels of cytokines including interleukin (IL) 1, IL2, interferon gamma, IL4, and IL10 were determined by ELISA. The mean change from baseline to 24 weeks of therapy was analysed. RESULTS: The two groups (DHEA n = 15; placebo n = 15) were well balanced for baseline characteristics. Only IL1beta and IL10 could be detected in the serum of lupus patients; however, there was no significant mean (SD) difference in serum IL1beta before and after treatment (9.94 (8.92) v 9.20 (6.49) pg/ml). IL10 demonstrated a greater and significant reduction from baseline (9.21 (9.66) to 1.89 (1.47) pg/ml in the DHEA treatment group). CONCLUSIONS: In a 24 week study of adult Chinese women with mild to moderate SLE, treatment with DHEA 200 mg once daily resulted in significant reduction of serum levels of IL10. This finding may suggest why DHEA could significantly reduce lupus flares. PMID- 15547087 TI - The myth of pulmonary Raynaud's phenomenon: the contribution of pulmonary arterial vasospasm in patients with systemic sclerosis related pulmonary arterial hypertension. AB - OBJECTIVE: To investigate the contribution of cold induced pulmonary vasospasm by peripheral and central cold stimulus in exacerbating pulmonary arterial hypertension (PAH) in patients with systemic sclerosis undergoing cardiac catheterisation. METHODS: In a prospective pilot study, 21 patients with systemic sclerosis and catheter proven PAH had mean pulmonary artery pressure (mPAP), pulmonary vascular resistance (PVR), and cardiac output (CO) measured before and after peripheral (hand immersion into cold water at 10-15 degrees C for two minutes if tolerated) and central (direct cold water at 4 degrees C injected into the right atrium) cold pressor challenge. Markers of endothelial activation, platelet function, and nitric oxide degradation were measured in blood sampled from the pulmonary artery. RESULTS: 19 of the patients (mean (SD) age, 56 (4) years; baseline mPAP, 34 (8) mm Hg; PVR, 420 (87) dyne.s.cm(-5); CO, 6.4 (1.8) l/min) tolerated cold hand immersion for the maximum two minute duration. All 21 tolerated central cold pressor challenge (three to five injections of 10 ml saline boluses at 4 degrees C). There was no significant change in haemodynamics after cold challenge by either route of provocation. Levels of endothelin-1, von Willebrand factor, fibrinogen, and 3-nitrotyrosine were raised compared with control values in patients with systemic sclerosis but without PAH, but did not change significantly after peripheral cold challenge. CONCLUSIONS: Pulmonary vasospasm in response to peripheral and centrally administered cold pressor challenge is unlikely to contribute to persistence of pulmonary arterial hypertension in patients with systemic sclerosis. PMID- 15547088 TI - Leflunomide inhibits transendothelial migration of peripheral blood mononuclear cells. AB - OBJECTIVES: To test whether the active metabolite of leflunomide (LEF-M), in addition to blocking the proliferation of activated lymphocytes by inhibiting dihydro-orotate dehydrogenase (DHODH), influences the transendothelial migration (TEM) of peripheral blood mononuclear cells (PBMC). METHODS: In an in vitro model of PBMC transmigration through an endothelial cell (EC) barrier, PBMC were re collected in three groups: cells not adherent to the EC, cells bound to, and cells which had migrated through, the EC layer. Experiments in which cells were pretreated with LEF-M (in the absence or in the presence of uridine) were compared with parallel experiments in the presence of medium alone. RESULTS: Preincubation of EC with LEF-M led to a 36 (SEM 16)% reduction in PBMC TEM (p<0.05). Likewise, preincubation of PBMC induced a reduction in their TEM of 39 (9)% (p<0.005). Incubation of both PBMC and EC with LEF-M had an additive effect (mean reduction of 48 (6)%, p<0.005). Incubation of PBMC with LEF-M also decreased monocytic CD44 expression (p<0.005) and PBMC-hyaluronan binding (p<0.05). Incubation of cells with LEF-M and uridine in addition to LEF-M reversed the inhibition of migration, suggesting that the observed effects were due to DHODH inhibition. Fluorocytometric analysis of PBMC subsets within the migrated population showed a decrease of monocytes, but not of B or T cells, after LEF-M treatment. CONCLUSIONS: LEF-M reduces monocytic adhesion molecule expression and TEM and may thus interfere with monocyte and EC activities in RA. Thus, the clinical effects of leflunomide may, at least in part, be due to blocking cell traffic into the inflamed synovia. PMID- 15547089 TI - Association of HLA-DM polymorphism with the production of antiphospholipid antibodies. AB - OBJECTIVE: To investigate whether variation in the HLA-DM gene is important in producing a group of pathogenic autoantibodies-antiphospholipid antibodies (aPL) on the basis that HLA class II restricted antigen presentation is involved in the production of aPL. METHODS: HLA-DMA and DMB polymorphisms were genotyped by polymerase chain reaction combined with restriction enzyme digestion in 51 white patients with primary antiphospholipid syndrome (APS), 82 with systemic lupus erythematosus (SLE) (42 with APS and 40 without APS), and 109 healthy white controls. The association with the aPL profile was examined. RESULTS: The distribution of DMA alleles in APS patients and in patients with APS associated with SLE was significantly different from that in controls by 4x2 chi(2) test with 3 degrees of freedom (p = 0.035 and 0.011, respectively), but it was not different between SLE patients without APS and controls. The allelic distribution of DMA was also different between patients with IgG class anticardiolipin antibody or those with lupus anticoagulant (LA) and controls (p = 0.012 and 0.007, respectively) and between patients with and without LA among SLE patients (p = 0.035). All these differences included the increase in DMA*0102 in the former groups. CONCLUSIONS: The results suggest that HLA-DMA*0102 or its linked gene(s) form one of the genetic risks for the production of aPL. PMID- 15547091 TI - Sensitivity to change of AIMS2 and AIMS2-SF components in comparison to M-HAQ and VAS-pain. AB - OBJECTIVE: To examine sensitivity to change of Dutch versions of AIMS2 (arthritis impact measurement scales-2) and AIMS2-SF (short form) components, in comparison with M-HAQ (modified health assessment questionnaire) and the 100 mm visual analogue scale for pain (VAS-pain) in patients with rheumatoid arthritis. METHODS: 218 patients participated in a study on patient education. Participants completed the Dutch AIMS2, M-HAQ, and VAS-pain at baseline and after one year; 165 completed both assessments. The education programme did not have any effect on health status. Patients were classified according to change over one year in their responses to the AIMS2 question about general health perception: improved health (n = 32), no change (n = 101), and poorer health (n = 32). Changes in scores over one year were tested with paired t tests, and standardised response means were calculated for AIMS2 and AIMS2-SF components, M-HAQ total score, and VAS-pain in the three classifications of change in health perception. RESULTS: AIMS2 and AIMS2-SF physical, symptom, and affect components showed similar sensitivity to change. The physical and symptom components performed better than M-HAQ and VAS-pain. AIMS2 and AIMS2-SF social interaction and role components were not sensitive to changes in general health perception. The role component was only applicable in 63 patients, because the others were unemployed, disabled, or retired. CONCLUSIONS: AIMS2-SF is a good alternative to the AIMS2 long form for the assessment of health status in rheumatoid arthritis, and is preferable to M-HAQ and VAS-pain. Use of the AIMS2-SF makes it easier and less costly to collect data and reduces the burden on patients. PMID- 15547090 TI - Long term effectiveness of intravenous immunoglobulin in Churg-Strauss syndrome. AB - OBJECTIVE: To study the long term effectiveness of intravenous immunoglobulin and plasmapheresis associated with prednisone and cyclophosphamide in Churg-Strauss syndrome. SUBJECTS: and methods: We studied 18 subjects with new onset Churg Strauss syndrome. All received the "standard" treatment based on prednisone (1 mg/kg/day for 1 month and then slowly tapered) and cyclophosphamide (2 mg/kg/day for 6 months in severe cases). In nine patients, synchronised cycles with plasmapheresis and intravenous immunoglobulin (2 g/kg) were repeated monthly for 6 months and every other month for a further three cycles. Clinical (disease activity monitored by Birmingham vasculitis activity score (BVAS) and damage index (modified Rankin score)) and functional (C reactive protein, blood eosinophil count, and electromyogram-electoneurogram) parameters were collected during treatment and the 3 year follow up period. RESULTS: After 12 months, all patients in the treatment group and four (44%) in the control group were in remission. At the end of the 3 year follow up period, we documented significant differences in BVAS (p<0.01), global damage (p<0.02), modified Rankin score (p<0.04), and the daily maintenance prednisone dose (p<0.002) between the two groups. We found a tendency towards lower frequency of relapse and incidence of osteoporosis in the treatment group. CONCLUSION: Complete clinical and functional recovery with a long term stable remission and a low incidence of side effects can be achieved by intravenous immunoglobulin associated with plasmapheresis in patients with Churg-Strauss syndrome. PMID- 15547092 TI - Expression of matrix metalloproteinases in patients with Wegener's granulomatosis. AB - BACKGROUND: Enhanced activity of matrix metalloproteinases (MMPs) has been reported to have a pathogenic role in several diseases such as cancer and cardiovascular disorders, and seems also to play a part in certain autoimmune diseases. OBJECTIVE: To examine whether enhanced MMP activity may also have a role in the pathogenesis of Wegener's granulomatosis (WG). METHODS: In a study group of 15 patients with WG and 15 controls, plasma levels and gene expression were measured in freshly isolated peripheral blood mononuclear cells (PBMCs) of several MMPs and their endogenous inhibitors (that is, tissue inhibitors of metalloproteinases (TIMPs)) by enzyme immunoassays and RNase protection assay, respectively. RESULTS: Whereas patients with WG in remission had enhanced gene expression of several MMPs and TIMPs in PBMCs, those with active disease had a selective up regulation of MMP-2 and MMP-8 compared with healthy controls, and a down regulation of TIMP-1 and TIMP-3 compared with other patients with WG. Moreover, plasma levels of TIMP-1 and MMP-8 correlated significantly with C reactive protein levels, further supporting an association between activation of the MMP/TIMP system and disease activity in WG. Finally, these changes in MMP/TIMP expression in WG were accompanied by increased total MMP activity in PBMC supernatants, particularly in those with active disease, suggesting a matrix degrading net effect. CONCLUSION: These findings suggest that disturbed MMP and TIMP activity has a role in the pathogenesis of WG. PMID- 15547093 TI - Impact of anti-tumour necrosis factor alpha treatment on admissions to hospital and days of sick leave in patients with ankylosing spondylitis. AB - OBJECTIVE: To analyse the impact of infliximab treatment on the number of hospital inpatient days and days of sick leave in patients with active ankylosing spondylitis (AS). METHODS: The data of a 2 year open extension study of a 12 week, double blind, randomised, placebo controlled trial, in which all patients with AS were treated with 5 mg/kg infliximab, were used to investigate the effect of anti-TNF treatment on admissions to hospital and days of sick leave. All patients were interviewed at baseline and at regular intervals during the study to collect this information by questionnaires. Patients who completed 2 years of treatment (n = 49) and those who did not (n = 20) were analysed separately. Sick leave analysis was restricted to currently employed patients (n = 38). RESULTS: During the 12 months before the screening visit, 20/49 (41%) completers had been admitted to hospital. After 1 and 2 years of treatment this percentage was reduced to 5/49 (10%; p<0.01), corresponding to a significant decrease in the mean number of inpatient days: from 11.1 to 0.6 after 1 year (p<0.01) and 2.9 after 2 years (p<0.01), respectively. No changes were seen in the dropout group. CONCLUSION: The use of infliximab in patients with active AS reduces some important costs of AS, but additional studies with detailed cost calculations are needed. PMID- 15547094 TI - Alpha beta but not gamma delta T cell clones in synovial fluids of patients with reactive arthritis show active transcription of tumour necrosis factor alpha and interferon gamma. AB - OBJECTIVE: To compare the cytokine expression profile of three CD8+, three CD4+, and three gammadelta+ T cell clones all derived from the synovial fluids of three patients with reactive arthritis (ReA). METHODS: Complementary DNA based microarrays containing the specific sequence of 56 cytokine transcripts were used for screening. Selected genes were confirmed by reverse transcriptase-polymerase chain reaction assay. RESULTS: Microarray showed that transcripts encoding for interferon gamma and tumour necrosis factor alpha were expressed by all CD8+ and CD4+ T cell clones. However, gammadelta+ T cells predominantly expressed transforming growth factor beta2 and granulocyte monocyte-colony stimulating factor. CONCLUSION: T lymphocyte clones from the joint of patients with ReA exhibit differential cytokine expression profiles. CD8+ and CD4+ T cells demonstrate a Th1 mediated profile, whereas gammadelta+ T cells show a more heterogeneous and less proinflammatory Th3 driven pattern. PMID- 15547095 TI - Telomerase activity in B and T lymphocytes of patients with systemic lupus erythematosus. AB - OBJECTIVE: To evaluate telomerase activity as a marker of lymphocyte proliferation in systemic lupus erythematosus (SLE). METHODS: CD19+, CD4+, and CD8+ lymphocytes were isolated from the peripheral blood of nine patients with SLE and nine healthy controls by means of magnetic bead-coupled antibodies and tested for telomerase activity with the TRAP assay. RESULTS: Telomerase activity was significantly increased in CD19+ B cells from patients with SLE. CD4+ and CD8+ T cells from lupus patients displayed increased mean telomerase activity, although the difference from normal controls did not reach statistical significance. CONCLUSIONS: Increased telomerase activity in the B and the T cell lineage might indicate activation and proliferation of these lymphocytes. PMID- 15547096 TI - Self assessment in continuous professional development: a valuable tool for individual physicians and scientific societies. AB - OBJECTIVES: To assess the appreciation of self assessment (SA) by multiple choice question (MCQ) tests during continuous professional development (CPD) meetings and to evaluate the attitude towards the option of recertification. METHODS: A questionnaire was completed at a meeting of the Swiss Societies of Rheumatology and Physical Medicine and Rehabilitation in 2003 (response rate 94.9% (223/235)). RESULTS: 60.9% of members found SA useful to assess rheumatological knowledge by MCQ tests; 71.3% thought it motivating to receive an anonymous feedback; 47.2% wanted an additional individualised feedback; 70.8% asked for SA to be continued every 2 years during CPD sessions; 26.3% favoured the option of recertification with identical standards to the Swiss certifying examination in rheumatology. Physicians in private practice less often chose the option of recertification than physicians employed by hospitals (OR = 2.09, 95% CI 1.03 to 4.18). No correlations between the type and duration of specialisation, personal reading time, frequency of attendance at CPD meetings, sex of the members, and the choices made were found. CONCLUSIONS: SA by MCQ tests during CPD meetings is highly accepted by Swiss rheumatologists. This reliable, valid, and economic method of learning needs assessment enables individual physicians and scientific societies to plan educational with specific goals in mind. PMID- 15547097 TI - Diagnostic accuracy of anti-cyclic citrullinated peptide antibodies in juvenile idiopathic arthritis. AB - OBJECTIVE: To correlate serum anti-cyclic citrullinated peptide antibodies (anti CCP) levels with juvenile idiopathic arthritis (JIA) subtypes and with an erosive disease course. METHODS: The study group comprised 122 children with JIA; 16 were evaluated during both active disease and remission. Nineteen children with systemic lupus erythematosus (SLE), 27 with rheumatoid arthritis (RA), and 15 healthy children were also included in the study. Twelve children with JIA were rheumatoid factor (RF) positive, and 34 patients had persistent erosive joint disease. Anti-CCP antibody levels were determined by ELISA; values above 5 relative units were regarded as positive. RESULTS: Three girls with seropositive polyarticular JIA and erosive joint disease had positive anti-CCP values. Children evaluated during active disease and remission, patients with SLE, and healthy children all had negative anti-CCP antibody levels. However, 19/27 (70%) adult patients with RA had positive anti-CCP antibody values. CONCLUSIONS: In contrast with RA, anti-CCP positivity is only rarely found in patients with JIA. In patients with RF positivity and/or in patients with erosive joint disease, anti-CCP can be detected. PMID- 15547099 TI - Cerebral venous thrombosis is associated with major vessel disease in Behcet's syndrome. AB - OBJECTIVE: To examine the association between the type of central nervous system (CNS) involvement (parenchymal disease and cerebral venous thrombosis (CVT)) and extra-cranial large vessel events, mainly venous thrombosis, in Behcet's syndrome (BS). METHODS: Conventional venous angiograms, Doppler ultrasonography, computed tomography, and MR angiography were used to study 88 patients with BS, with (n = 88) and without (n = 80) CNS disease for the presence of major vessel disease. RESULTS: Major vessel involvement among the male patients with and without CNS disease (21/73 (29%) v 18/80 (23%), respectively) showed no significant differences (p = 0.374). When patients with CNS disease were stratified according to the type of CNS involvement, 7/11 (64%) patients with CVT had major vessel disease compared with 15/77 (19%) patients with parenchymal disease (p = 0.004). The mean (SD) age of onset of CVT (23.1 (8.8) years) among the male patients was significantly earlier than among the men with parenchymal disease (32.0 (7.5); p = 0.002). CONCLUSIONS: CVT in BS was strongly associated with peripheral major vessel disease and occurred earlier in the disease course than the parenchymal type of CNS disease. As superficial thrombophlebitis also occurs more frequently in patients with major vessel disease in BS, this may suggest a common pathogenic mechanism. PMID- 15547098 TI - Stress fractures in rheumatoid arthritis: a case series and case-control study. AB - OBJECTIVE: To review cases of stress fracture in patients with rheumatoid arthritis (RA) presenting to our service over a 10 year period; to identify possible risk factors and test these in a case-control study. METHODS: A retrospective case note review of all patients with a final diagnosis of stress fracture, presenting between 1990 and 1999 was performed. A case-control study of consecutive patients with RA, matched for age, sex and duration of disease, attending the same clinics. RESULTS: CASE SERIES: 24 stress fractures were identified in 18 patients, representing 0.8% of the RA clinic population; all were women, median age 69.5 years (range 47-79). Bone mineral densitometry showed median T scores of -3.06 and -2.27 SD at the hip and lumbar spine, respectively. Case-control study: In the stress fractures group, past steroid doses were higher (p = 0.003). No significant differences in the bone mineral density (BMD) T score at the hip or lumbar spine were found (p = 0.59, p = 0.77). CONCLUSIONS: Stress fractures are a significant cause of morbidity in RA. Diagnosis is often delayed and presentation can be misleading. Past steroid use, particularly at higher doses, confers an increased risk of stress fracture, but the increased risk is not attributable to osteoporosis as assessed by BMD. PMID- 15547100 TI - DNA mismatch repair enzyme expression in synovial tissue. AB - BACKGROUND: Oxidative stress in RA synovial tissue can cause DNA damage and suppress the DNA mismatch repair (MMR) system in cultured synoviocytes. This mechanism includes two enzyme complexes, hMutSalpha (hMSH2/hMSH6) and hMutSbeta (hMSH2/hMSH3). OBJECTIVE: To examine the expression and distribution of MMR enzymes in synovial tissues from patients with arthritis and from normal subjects. METHODS: Synovial tissues from patients with RA, osteoarthritis (OA), or normal subjects were analysed by immunohistochemistry using monoclonal antibodies to hMSH2, hMSH3, and hMSH6. MMR protein expression was evaluated by computer assisted digital image analysis. RESULTS: hMSH2, hMSH3, and hMSH6 were found in most synovial tissues evaluated, with greater levels in the intimal lining than sublining regions. In RA and OA, sublining perivascular staining for hMSH6 and hMSH3 was also prominent. Significantly higher sublining expression of hMSH2, hMSH3, and hMSH6 was seen in RA and OA than in normal synovium. Double label immunohistochemistry demonstrated that the main cells expressing MMR enzymes were CD68(+) and CD68(-) cells in the intimal lining. CONCLUSIONS: DNA MMR enzyme expression is greatest in the synovial intimal lining layer, where maximal oxidative stress in RA occurs. Although MMR enzyme expression is greater in RA than in normal tissue, this compensatory response cannot overcome the genotoxic environment, and DNA damage accumulates. PMID- 15547101 TI - A need for greater reporting of socioeconomic status and race in clinical trials. PMID- 15547102 TI - Sinusoidal dilatation: a rare side effect of azathioprine. PMID- 15547103 TI - Tick bite fever and arthritis associated with travel to Africa. PMID- 15547104 TI - Extracorporeal shock wave therapy in the treatment of inferior calcaneal enthesophytosis: outcome by fan-beam dual x ray absorptiometry (DXA). PMID- 15547105 TI - Clinical predictors of fetal and maternal outcome in Chinese patients with systemic lupus erythematosus. PMID- 15547106 TI - Temporomandibular joint pseudogout: an uncommon site for a familiar condition. PMID- 15547107 TI - Antinuclear antibodies are common in an infectious environment but do not predict systemic lupus erythematosus. PMID- 15547108 TI - Metacarpophalangeal joint of short metacarpal bone in rheumatoid arthritis. PMID- 15547109 TI - European recommendations for musculoskeletal pain. PMID- 15547110 TI - Cyclooxygenase-2 inhibition improves vascular endothelial dysfunction in a rat model of endotoxic shock: role of inducible nitric-oxide synthase and oxidative stress. AB - We investigated whether cyclooxygenase (COX) isoforms (COX-1 and COX-2) and decreased NO availability contribute to endothelial dysfunction in endotoxemic rats. The involvement of reactive oxygen species (ROS) was also evaluated. Rats were injected with Salmonella-derived lipopolysaccharide or saline. After 6 h, endothelial function of mesenteric resistance arteries was evaluated. In controls, acetylcholine (ACh)-induced relaxation was inhibited by the nitric oxide synthase inhibitor N(G)-monomethyl-l-arginine (l-NMMA) and unaffected by 5,5-dimethyl-3-(3-fluorophenyl)-4-(4-methylsulphonyl)-phenyl-2(5H)-furanone (DFU) (COX-2 inhibitor). In lipopolysaccharide (LPS)-treated rats, the response to ACh was blunted compared with controls, less sensitive to l-NMMA, and enhanced by DFU. COX-2 blockade also improved the inhibitory effect of l-NMMA on cholinergic relaxation. SC-560 [5-(4-clorophenyl)-1-(4-metoxyphenyl)-3 trifluoromethylpirazole] (COX-1 inhibitor) did not modify the response to ACh in both groups. LPS-induced endothelial dysfunction was unaffected by the thromboxane A(2) (TxA(2)) receptor antagonist SQ-29548 (7-[3-[[2 [(phenylamino)carbonyl]hydrazino]methyl]-7-oxabicyclo[2.2.1] hept-2-yl] [1S(1alpha,2alpha(Z),3alpha,4alpha)]-5-heptenoic acid). In vivo inducible nitric oxide synthase (iNOS) inhibition by S-methylisothiourea partly attenuated LPS induced endothelial dysfunction. The antioxidants ascorbic acid and superoxide dismutase normalized endothelium-dependent relaxation and restored the inhibitory action of l-NMMA on ACh. Responses to sodium nitroprusside were similar in both groups. In LPS-treated rats, reverse transcription-polymerase chain reaction showed a marked increase in mesenteric iNOS and COX-2 expressions, whereas endothelial nitric-oxide synthase and COX-1 were unchanged. LPS-induced COX-2 overexpression was reduced but not abrogated by S-methylisothiourea. LPS-induced COX-2 up-regulation was also documented by immunohistochemistry. In conclusion, mesenteric resistance vessels from endotoxemic rats show impaired endothelial function due to reduced NO availability, a condition that is partly ascribable to an iNOS-dependent enhanced COX-2 expression, whereas TxA(2) does not seem to be involved. Oxidative stress is the main mechanism responsible for reduced NO availability, and COX-2 might act as a source of ROS. PMID- 15547111 TI - 5-Aza-cytidine is a potent inhibitor of DNA methyltransferase 3a and induces apoptosis in HCT-116 colon cancer cells via Gadd45- and p53-dependent mechanisms. AB - Methyltransferase inhibitors commonly used in clinical trials promote tumor cell death, but their detailed cytotoxic action is not yet fully understood. A deeper knowledge about their apotosis-inducing mechanisms and their interaction with DNA methyltransferases (DNMTs) DNMT1, DNMT3a, and DNMT3b might allow the design of more effective drugs with lower cytotoxicity. 5-aza-cytidine (5-aza-CR), a potent inhibitor of DNMT1, is known to induce demethylation and reactivation of silenced genes. In this study, we investigated the p53 dependence of apoptotic, cell cycle, and growth inhibitory effects of 5-aza-CR, as well as the influence on the expression level of DNMT1, DNMT3a, and DNMT3b in the colon cancer cell line HCT 116. Exposure to 5-aza-CR induced the up-regulation of genes promoting cell cycle arrest and DNA repair (p21(WAF1) and GADD45) or apoptosis (p53, RIPK2, Bak1, caspase 5, and caspase 6). In parallel, there was a down-regulation of antiapoptotic Bcl2 protein and the G(2)/M-mediator cyclin B1. Co-incubation with pifithrin-alpha (PFT-alpha), a selective p53 inhibitor, restored GADD45, Bcl2, cyclin B1, and p21(WAF1) expression levels and almost completely reversed the growth inhibitory, cell cycle, and apoptotic effects of 5-aza-CR. 5-aza-CR treatment caused global demethylation and reactivation of p16(INK4) expression. There was a marked decrease in DNMT1 and DNMT3a mRNA expression, with PFT-alpha reversing these effects. However, 5-aza-CR treatment did not modulate DNMT3b expression. Our data demonstrate that 5-aza-CR action in HCT-116 is mediated by p53 and its downstream effectors p21(WAF1) and GADD45. This is the first report to show a link between p53 and regulation of DNMT1 and de novo methyltransferase DNMT3a. PMID- 15547112 TI - Nucleoside transport at the blood-testis barrier studied with primary-cultured sertoli cells. AB - Nucleosides are essential for nucleotide synthesis in testicular spermatogenesis. In the present study, the mechanism of the supply of nucleosides to the testicular system across the blood-testis barrier was studied using primary cultured Sertoli cells from rats and TM4 cells from mice. Uptake of uridine by these cells was time- and concentration-dependent. Uridine uptake was decreased under Na(+)-free conditions, and the system was presumed to be high affinity, indicating an Na(+)-dependent concentrative nucleoside transporter (CNT) is involved. On the other hand, nitrobenzylthioinosine, a potent inhibitor of Na(+) independent equilibrative nucleoside transporters (ENTs), inhibited uridine uptake by the Sertoli cells in a concentration-dependent manner. Expression of nucleoside transporters ENT1, ENT2, ENT3, CNT1, CNT2, and CNT3 was detected in Sertoli cells by reverse transcriptase-polymerase chain reaction analysis. Inhibition studies of the uptake of uridine by various nucleosides both in the presence and absence of Na(+) indicated that the most of those expressed nucleoside transporters, ENTs and CNTs, are involved functionally. These results demonstrated that Sertoli cells are equipped with multiple nucleoside transport systems, including ENT1, ENT2, and CNTs, to provide nucleosides for spermatogenesis. PMID- 15547113 TI - Activation of dopamine D1-like receptors induces acute internalization of the renal Na+/phosphate cotransporter NaPi-IIa in mouse kidney and OK cells. AB - The Na(+)/phosphate cotransporter NaPi-IIa (SLC34A1) is the major transporter mediating the reabsorption of P(i) in the proximal tubule. Expression and activity of NaPi-IIa is regulated by several factors, including parathyroid hormone, dopamine, metabolic acidosis, and dietary P(i) intake. Dopamine induces natriuresis and phosphaturia in vivo, and its actions on several Na(+) transporting systems such as NHE3 and Na(+)-K(+)-ATPase have been investigated in detail. Using freshly isolated mouse kidney slices, perfused proximal tubules, and cultured renal epithelial cells, we examined the acute effects of dopamine on NaPi-IIa expression and localization. Incubation of isolated kidney slices with the selective D(1)-like receptor agonists fenoldopam (10 microM) and SKF-38393 (10 microM) for 1 h induced NaPi-IIa internalization and reduced expression of NaPi-IIa in the brush border membrane (BBM). The D(2)-like selective agonist quinpirole (1 microM) had no effect. The D(1) and D(2) agonists did not affect the renal Na(+)/sulfate cotransporter NaSi in the BBM of the proximal tubule. Studies with isolated perfused proximal tubules demonstrated that activation of luminal, but not basolateral, D(1)-like receptors caused NaPi-IIa internalization. In kidney slices, inhibition of PKC (1 microM chelerythrine) or ERK1/2 (20 microM PD-098089) pathways did not prevent the fenoldopam-induced internalization. Inhibition with the PKA blocker H-89 (10 microM) abolished the effect of fenoldopam. Immunoblot demonstrated a reduction of NaPi-IIa protein in BBMs from kidney slices treated with fenoldopam. Incubation of opossum kidney cells transfected with NaPi-IIa-green fluorescent protein chimera shifted fluorescence from the apical membrane to an intracellular pool. In summary, dopamine induces internalization of NaPi-IIa by activation of luminal D(1)-like receptors, an effect that is mediated by PKA. PMID- 15547114 TI - L-type calcium channels in the renal microcirculatory response to endothelin. AB - The signaling pathways of endothelin (ET)-1-mediated vasoconstriction in the renal circulation have not been elucidated but appear to be distinct between ET(A) and ET(B) receptors. The purpose of this study was to determine the role of L-type Ca(2+) channels in the vasoconstrictor response to ET-1 and the ET(B) receptor agonist sarafotoxin 6c (S6c) in the rat kidney. Renal blood flow (RBF) was measured with an ultrasonic flow probe in anesthetized rats, and a microcatheter was inserted into the renal artery for drug infusion. All rats were given vehicle (0.9% NaCl) or three successive bolus injections (1, 10, and 100 pmol) of ET-1 or S6c at 30-min intervals (n = 6 in each group). ET-1 and S6c produced dose-dependent decreases in RBF. The Ca(2+) channel blocker nifedipine (1.5 mug) significantly attenuated the RBF response only at the highest doses of ET-1 and S6c. In the isolated blood-perfused juxtamedullary nephron preparation, Ca(2+) channel blockade with diltiazem had a very small inhibitory effect on ET-1 induced decreases in afferent arteriolar diameter only at the lowest concentrations of ET-1. In vascular smooth muscle cells isolated from preglomerular vessels, ET-1 produced a typical biphasic Ca(2+) response, whereas S6c had no effect on cytosolic Ca(2+). Furthermore, Ca(2+) channel blockade (diltiazem or Ni(2+)) had no effect on the peak or sustained increase in cytosolic Ca(2+) produced by ET-1. These results support the hypothesis that L type Ca(2+) channels play only a minor role in the constrictor responses to ET-1 in the renal microcirculation. PMID- 15547115 TI - Programming blood pressure in adult SHR by shifting perinatal balance of NO and reactive oxygen species toward NO: the inverted Barker phenomenon. AB - The "programming hypothesis" proposes that an adverse perinatal milieu leads to adaptation that translates into cardiovascular disease in adulthood. The balance between nitric oxide (NO) and reactive oxygen species (ROS) is disturbed in cardiovascular diseases, including hypertension. Conceivably, this balance is also disturbed in pregnancy, altering the fetal environment; however, effects of perinatal manipulation of NO and ROS on adult blood pressure (BP) are unknown. In spontaneously hypertensive rats (SHR), NO availability is decreased and ROS are increased compared with normotensive Wistar-Kyoto rats, and, despite the genetic predisposition, the perinatal environment can modulate adult BP. Our hypothesis is that a disturbed NO-ROS balance in the SHR dam persistently affects BP in her offspring. Dietary supplements, which support NO formation and scavenge ROS, administered during pregnancy and lactation resulted in persistently lower BP for up to 48 wk in SHR offspring. The NO donor molsidomine and the superoxide dismutase mimic tempol-induced comparable effects. Specific inhibition of inducible nitric oxide synthase (NOS) reduces BP in adult SHR, suggesting that inducible NOS is predominantly a source of ROS in SHR. Indeed, inducible NOS inhibition in SHR dams persistently reduced BP in adult offspring. Persistent reductions in BP were accompanied by prevention of proteinuria in aged SHR. We propose that in SHR the known increase in ANG II type 1 receptor density during development leads to superoxide production, which enhances inducible NOS activity. The relative shortage of substrate and cofactors leads to uncoupling of inducible NOS, resulting in superoxide production, activating transcription factors that subsequently again increase inducible NOS expression. This vicious circle probably is perpetuated into adult life. PMID- 15547116 TI - The role of NOS2 and NOS3 in renal protein and arginine metabolism during early endotoxemia in mice. AB - Previously, we observed an enhanced renal protein synthesis and increased de novo arginine production in the early response to endotoxemia in wild-type Swiss mice (Hallemeesch MM, Soeters PB, and Deutz NE. Am J Physiol Renal Physiol 282: F316 F323, 2002). To establish whether these changes are regulated by nitric oxide (NO) synthesized by NO synthase isoforms NOS2 and NOS3, we studied C57BL6/J wild type (WT), NOS2-deficient (NOS2(-/-)), and NOS3-deficient (NOS3(-/-)) mice under baseline (unstimulated) and LPS-treated conditions. The metabolism of renal protein, amino acid, and arginine was studied at the whole body level and across the kidney by infusing the stable isotopes l-[phenyl-(2)H(5)]phenylalanine, l [phenyl-(2)H(2)]tyrosine, l-guanidino-[(15)N(2)]arginine, and l-[ureido (13)C,(2)H(2)]citrulline. Renal blood flow was measured using radioactive PAH extraction. Under baseline conditions, renal blood flow was significantly reduced in NOS2(-/-) mice (0.29 +/- 0.01 vs. 0.48 +/- 0.07 ml.10 g body wt(-1).min(-1) in WT) (P < 0.05), and de novo arginine production was lower in NOS2(-/-) mice. After LPS challenge, renal protein turnover and arginine production increased in all three groups (P < 0.05), even though renal de novo arginine synthesis did not increase. The expected increase in renal citrulline production and disposal after LPS was not observed in NOS2(-/-) mice (P = 0.06). Collectively, these data show that NOS2 is constitutively expressed in the kidney and remarkably functional as it affects renal blood flow and de novo arginine production under baseline conditions and is important for the increase in renal citrulline turnover during endotoxemia. NOS3, in contrast, appears less important for renal metabolism. The increase in renal protein turnover during endotoxemia does not depend on NOS2 or NOS3 activity. PMID- 15547117 TI - Basolateral K+ conductance in principal cells of rat CCD. AB - Whole cell K+ current was measured by forming seals on the luminal membrane of principal cells in split-open rat cortical collecting ducts. The mean inward, Ba2+-sensitive conductance, with 40 mM extracellular K+, was 76 +/- 12 and 141 +/ 22 nS/cell for animals on control and high-K+ diets, respectively. The apical contribution to this was estimated to be 3 and 16 nS/cell on control and high-K+ diets, respectively. To isolate the basolateral component of whole cell current, we blocked ROMK channels with either tertiapin-Q or intracellular acidification to pH 6.6. The current was weakly inward rectifying when bath K+ was > or =40 mM but became more strongly rectified when bath K+ was lowered into the physiological range. Including 1 mM spermine in the pipette moderately increased rectification, but most of the outward current remained. The K+ current did not require intracellular Ca2+ and was not inhibited by 3 mM ATP in the pipette. The negative log of the acidic dissociation constant (pKa) was approximately 6.5. Block by extracellular Ba2+ was voltage dependent with apparent Ki at -40 and -80 mV of approximately 160 and approximately 80 microM, respectively. The conductance was TEA insensitive. Substitution of Rb+ or NH4+ for K+ led to permeability ratios of 0.65 +/- 0.07 and 0.15 +/- 0.02 and inward conductance ratios of 0.17 +/- 0.03 and 0.57 +/- 0.09, respectively. Analysis of Ba2+-induced noise, with 40 mM extracellular K+, yielded single-channel currents of 0.39 +/- 0.04 and -0.28 +/- 0.04 pA at voltages of 0 and -40 mV, respectively, and a single-channel conductance of 17 +/- 1 pS. PMID- 15547118 TI - Anatomy, biomechanics, and surgical approach to synovial folds within the joints of the foot. AB - The presence of synovial folds in various joints of the foot has been previously documented. The function and clinical significance of these structures within the joint have not been established. Histologically they are considered anatomically different from a meniscus primarily owing to their makeup of loose connective tissue with nerve fibrils and several synovial cell layers. We hypothesize that the function of these folds is similar to that of the menisci: to increase joint congruity and stability. We further hypothesize that these folds will be present in joints of the foot that require greater stability. To demonstrate this, 41 fixated cadaveric feet were sectioned in the sagittal plane and the incidence and locations of the synovial folds were documented. Three fixated cadaveric feet were evaluated using a materials testing machine. The first metatarsophalangeal joint was incised, and the presence of the synovial fold was documented. The joint was then taken through its range of motion with and without the synovial fold while data on the force and displacement were collected. The steps were then repeated for the ankle joint. The results showed statistically stiffer ankle and first metatarsophalangeal joints with the synovial fold present, as determined by the stress-strain curve. On the basis of the presence and location of these synovial folds, we demonstrated arthroscopic surgical approaches to many of the documented joints that contain these folds. Because the folds contain synovial cells and vascular tissue, damage to them can result in considerable pain. In such cases, arthroscopic surgery would be of benefit. Further research may indicate whether they need to be salvaged during joint procedures to facilitate normal joint function or should be removed to reduce postoperative complications. PMID- 15547119 TI - A comparison of results from two mycology laboratories for the diagnosis of onychomycosis: a study of 85 cases in a geriatric population. AB - An investigative study was performed to compare the results from two mycology laboratories for the diagnosis of onychomycosis in a geriatric population and to determine the possible pharmacologic treatments based on the two laboratories' results. In this study, 85 cases of suspected onychomycosis involving men and women 65 years and older from a nursing home setting in South Florida were used. Samples were taken from the hallux toenail and sent to two different mycology laboratories for fluorescent potassium hydroxide preparation and microscopic examination of a fungal culture. Of the 85 cases studied, the two mycology laboratories reported similar potassium hydroxide preparation results for 58.8% of the patients and similar fungal culture results for genus and species identification for 37.6% of the patients. When the potassium hydroxide preparation and fungal culture results were combined, the two mycology laboratories reported similar results for only 27.1% of the patients. As a result of the two mycology laboratories' findings, the possible US Food and Drug Administration-approved pharmacologic treatments may differ for 43.5% of the patients studied. The discrepancy between the two independent laboratories leaves physicians to question the reproducibility of fluorescent potassium hydroxide preparation and fungal culture analysis in a geriatric patient population for the diagnosis of onychomycosis. PMID- 15547121 TI - Effectiveness of different types of foot orthoses for the treatment of plantar fasciitis. AB - Clinicians often use foot orthoses to manage the symptoms of plantar fasciitis. Although there has been considerable research evaluating the effectiveness of orthoses for this condition, there is still a lack of scientific evidence that is of suitable quality to fully inform clinical practice. Randomized controlled trials are recognized as the "gold standard" when evaluating the effectiveness of treatments. We discuss why randomized controlled trials are so important, the features of a well-conducted randomized controlled trial, and some of the problems that arise when trial design is not sound. We then evaluate the available evidence for the use of foot orthoses, with particular focus on published randomized controlled trials. From the evidence to date, it seems that foot orthoses do have a role in the management of plantar fasciitis and that prefabricated orthoses are a worthwhile initial management strategy. At this time, however, it is not possible to recommend either prefabricated or customized orthoses as being better, and it cannot be inferred that customized orthoses are more effective over time and therefore have a cost advantage. Additional good quality randomized controlled trials are needed to answer these questions. PMID- 15547120 TI - Associations between static and functional measures of joint function in the foot and ankle. AB - Clinicians have traditionally assessed range of motion of the first metatarsophalangeal and ankle joints in a static position. It is unclear, however, if these measurements accurately reflect functional sagittal plane limitations of these joints during gait. For 50 patients (100 feet), we assessed available dorsiflexion at the first metatarsophalangeal and ankle joints, as well as the presence of pinch callus. We then compared these findings with 11 functional gait parameters, as measured using a pressure sensor system. After adjusting for age, weight, smoking status, glycosylated hemoglobin, and insensitivity to monofilament, we found that patients with pinch callus demonstrated statistically significant compensatory gait patterns in 7 of 11 measures. Hallux limitus and equinus patients demonstrated six and three statistically significant associations, respectively. Pinch callus seems to be as predictive of functional gait alterations as static first metatarsophalangeal joint and ankle dorsiflexion. PMID- 15547122 TI - The histologic host response to liquid silicone injections for prevention of pressure-related ulcers of the foot: a 38-year study. AB - This study analyzed the histologic effects of and host response to subdermally injected liquid silicone to augment soft-tissue cushioning of the bony prominences of the foot. A total of 148 postmortem and surgical specimens of pedal skin with attached soft tissue were obtained from 49 patients between July 1, 1974, and November 30, 2002. The longest period that silicone was in vivo was 38 years. The specimens were then processed into paraffin blocks and examined for specific findings. The variables considered included distribution of silicone within the tissue, host response, migration to regional lymph nodes, and viability of the host tissue after treatment. The host response to silicone therapy consisted primarily of delicate-to-robust fibrous deposition and histiocytic phagocytosis, with eventual formation of well-formed elliptic fibrous pads. The response in the foot appears different from that in the breast and other areas of the body previously studied. No examples of granulomas, chronic lymphoplasmacytic inflammation, or granulation tissue formation were seen, with only rare foreign-body giant cells present. Silicone injections in fat pads for the treatment of atrophy and loss of viable tissue show a histologically stable and biologically tolerated host response that is effective, with no evidence of any systemic changes. PMID- 15547123 TI - The effect of 5-degree valgus and varus rearfoot wedging on peak hallux dorsiflexion during gait. AB - The dynamic effects of 5 degrees varus and valgus rearfoot wedging on peak hallux dorsiflexion were investigated in 30 asymptomatic subjects (5 males and 25 females). Statistically significant reductions in peak hallux dorsiflexion were found with rearfoot varus wedging and rearfoot valgus wedging. Furthermore, the reduction in peak hallux dorsiflexion occurring with rearfoot varus wedging was statistically significant compared with that associated with rearfoot valgus wedging. These findings have implications for the orthotic management of a variety of lower-limb pathologies. PMID- 15547124 TI - Low-voltage direct current as a fungicidal agent for treating onychomycosis. AB - Onychomycosis, most commonly caused by two species of dermatophyte fungi- Trichophyton rubrum and Trichophyton mentagrophytes--is primarily treated with regimens of topical and systemic antifungal medications. This study was undertaken to evaluate in vitro the efficacy of low-voltage direct current as an antifungal agent for treating onychomycosis. Agar plate cultures of T rubrum and T mentagrophytes were subjected to low-voltage direct current electrostimulation, and antifungal effects were observed as zones in the agar around the electrodes lacking fungal growth. Zones devoid of fungal growth were observed for T rubrum and T mentagrophytes around anodes and cathodes in a dose-dependent manner in the current range of 500 microA to 3 mA. Low-voltage direct current electrostimulation has great clinical potential for the treatment of onychomycosis and perhaps other superficial maladies of fungal etiology. PMID- 15547125 TI - Measurement of foot dorsiflexion: a modified Lidcombe template. AB - The Lidcombe template was introduced in 1991 for the nonweightbearing assessment of ankle joint dorsiflexion, and it has shown excellent reliability in impaired and unimpaired adult populations. We discuss limitations of the original template and test the reliability of a modified apparatus in an adolescent population. Intrarater and interrater reliability were assessed for 14 children (28 limbs) aged 7 to 14 years, returning intraclass correlation coefficient (1,1) results of greater than 0.99 for both aspects of reliability. PMID- 15547126 TI - Reconstruction of the great toe ski-slope, sunken-nail deformity with a buried adipofascial flap. AB - Development of a ski-slope deformity following loss of the great toe nail plate is a problematic condition with few conservative or surgical options available. The condition becomes more difficult to treat when the distal, medial, and lateral labial nail folds are hypertrophied, creating the appearance of a sunken nail. We present a case of ski-slope, sunken-nail deformity following multiple attempts at chemical nail matrixectomy. The patient's persistent pain and deformity were managed through 1) nail plate avulsion and complete surgical excision of the germinal nail matrix, 2) remodeling of the distal phalanx, and 3) elevation of an adipofascial flap from the plantar tuft of the great toe, which was brought from plantar to dorsal and interposed between the dorsal aspect of the distal phalanx and the overlying nail bed in buried fashion. The combination of these procedures elevated the nail bed, which restored normal architecture to the great toe and relieved the pain associated with the chronic deformity. This case demonstrates a potential complication of a commonly performed procedure and a salvage technique useful for dealing with the resultant ski-slope, sunken-nail deformity. PMID- 15547127 TI - Norwegian scabies in the immunocompromised patient. AB - Norwegian, or crusted, scabies can be defined as a generalized severe scabies (Sarcoptes scabiei var. hominis) infestation usually affecting the immunocompromised patient that is most commonly seen with the leukemia-lymphoma group of neoplasms. The diagnosis is commonly missed, which can lead to mismanagement. We describe a patient with Norwegian scabies involving the lower extremities. The patient circumstances and treatment, as well as a review of the literature, are presented. The diagnosis of scabies should always be considered in patients with advanced malignancies and associated pruritus. PMID- 15547128 TI - Accessory soleus muscle. AB - Accessory soleus muscle is an uncommon anatomical variant that may present as a soft-tissue mass in the posteromedial region of the ankle. It is congenital in origin but usually presents in the second or third decade of life. Although it is a rare entity, accessory soleus muscle should be included in the differential diagnosis of soft-tissue swelling of the ankle. Awareness of the clinical presentation and specific findings of computed tomography, magnetic resonance imaging, and electromyography help with diagnosis without surgical exploration. We describe a 30-year-old patient with accessory soleus muscle. Magnetic resonance imaging features of the case are described, and the literature is briefly reviewed. PMID- 15547129 TI - Treatment of Freiberg's infraction with the titanium hemi-implant. AB - Freiberg's infraction is a relatively rare disease for which there is currently no consensus regarding surgical management. We present a case study describing a surgical procedure that uses a novel metatarsophalangeal hemi-implant. This procedure does not alter the metatarsal parabola, and it allows for other surgical procedures to be performed in the future. PMID- 15547130 TI - Transfer of the peroneus brevis tendon after resection of the fifth metatarsal base. AB - After resection of bone or amputation, postoperative stump breakdown occurs frequently. Furthermore, the altered mechanics with ambulation are difficult to control with bracing and orthoses alone. During the past 10 years, the peroneus brevis tendon has been transferred to various locations in the foot after resection of the fifth metatarsal base in an effort to provide continued balance between the supinatory and pronatory forces needed for a steady gait. In patients who have had a peroneus brevis tendon transfer, the rate of postoperative ulceration and the need for further bony resection is minimal. Analysis of the biomechanical influences and effects of different anatomical placements of the transferred tendon reveals the importance of transfer of the peroneus brevis tendon. PMID- 15547131 TI - American Podiatric Medical Association Missing Time Survey, 2004. AB - A 2004 survey of US adults found that 19% had experienced foot problems at work at some time. As a result, 38% reported lost productivity and 28% missed time at work. Younger, less educated male workers were more likely to suffer from foot problems. The percentage of the total population surveyed who missed time at work owing to foot problems was 5.4% in 2004. In a previous survey conducted in 2000, the corresponding percentage was 6.6%. PMID- 15547132 TI - The driving force for molecular evolution of translation. AB - It is widely argued that protein synthesis evolved out of an RNA world, in which catalytic and other biological functions now carried out by proteins were performed by RNAs. However, it is not clear what selective advantage would have provided the driving force for evolution of a primitive translation apparatus, because of the unlikelihood that rudimentary polypeptides would have contributed sufficiently useful biological functions. Here, I suggest that the availability of even simple peptides could have significantly enlarged the otherwise limited structure space of RNA. In other words, translation initially evolved not to create a protein world, but to extend the structural, and therefore the functional, capabilities of the RNA world. Observed examples of substantial structural rearrangements in RNA that are induced by binding of peptides and other small molecules support this possibility. PMID- 15547133 TI - RNA kink turns to the left and to the right. AB - A helix-loop-helix within the group I intron has most of the canonical sequence elements of a kink turn (K-turn), yet it bends in the opposite direction. The reverse K-turn kinks toward the major rather than the minor grooves of the flanking helices. This suggests that there are two distinct subclasses of tertiary structures that a K-turn secondary structure can adopt. The final structure may be specified by external factors, such as protein binding or the tertiary structural context, rather than the intrinsic conformation of the RNA. PMID- 15547134 TI - Crystal structure of a group I intron splicing intermediate. AB - A recently reported crystal structure of an intact bacterial group I self splicing intron in complex with both its exons provided the first molecular view into the mechanism of RNA splicing. This intron structure, which was trapped in the state prior to the exon ligation reaction, also reveals the architecture of a complex RNA fold. The majority of the intron is contained within three internally stacked, but sequence discontinuous, helical domains. Here the tertiary hydrogen bonding and stacking interactions between the domains, and the single-stranded joiner segments that bridge between them, are fully described. Features of the structure include: (1) A pseudoknot belt that circumscribes the molecule at its longitudinal midpoint; (2) two tetraloop-tetraloop receptor motifs at the peripheral edges of the structure; (3) an extensive minor groove triplex between the paired and joiner segments, P6-J6/6a and P3-J3/4, which provides the major interaction interface between the intron's two primary domains (P4-P6 and P3 P9.0); (4) a six-nucleotide J8/7 single stranded element that adopts a mu-shaped structure and twists through the active site, making critical contacts to all three helical domains; and (5) an extensive base stacking architecture that realizes 90% of all possible stacking interactions. The intron structure was validated by hydroxyl radical footprinting, where strong correlation was observed between experimental and predicted solvent accessibility. Models of the pre-first and pre-second steps of intron splicing are proposed with full-sized tRNA exons. They suggest that the tRNA undergoes substantial angular motion relative to the intron between the two steps of splicing. PMID- 15547135 TI - A synthetic A tail rescues yeast nuclear accumulation of a ribozyme-terminated transcript. AB - To investigate the role of 3' end formation in yeast mRNA export, we replaced the mRNA cleavage and polyadenylation signal with a self-cleaving hammerhead ribozyme element. The resulting RNA is unadenylated and accumulates near its site of synthesis. Nonetheless, a significant fraction of this RNA reaches the cytoplasm. Nuclear accumulation was relieved by insertion of a stretch of DNA-encoded adenosine residues immediately upstream of the ribozyme element (a synthetic A tail). This indicates that a 3' stretch of adenosines can promote export, independently of cleavage and polyadenylation. We further show that a synthetic A tail-containing RNA is unaffected in 3' end formation mutant strains, in which a normally cleaved and polyadenylated RNA accumulates within nuclei. Our results support a model in which a polyA tail contributes to efficient mRNA progression away from the gene, most likely through the action of the yeast polyA-tail binding protein Pab1p. PMID- 15547136 TI - Nonsense-mediated decay does not occur within the yeast nucleus. AB - Nonsense-mediated decay (NMD) is a eukaryotic regulatory process that degrades mRNAs with premature termination codons (PTCs). Although NMD is a translation dependent process, there is evidence from mammalian systems that PTC recognition and mRNA degradation takes place in association with nuclei. Consistent with this notion, degradation of mammalian PTC-containing mRNAs occurs when they are bound by the cap binding complex (CBC) during a "pioneer" round of translation. Moreover, there are reports indicating that a PTC can trigger other nuclear events such as alternative splicing, abnormal 3' end processing, and accumulation of pre-mRNA at transcription sites. To examine whether a PTC can elicit similar nuclear events in yeast, we used RNA export-defective mutants to sequester mRNAs within nuclei. The results indicate that nuclear PTC-containing yeast RNAs are NMD insensitive. We also observed by fluorescent in situ hybridization that there was no PTC effect on mRNA accumulated at the site of transcription. Finally, we show that yeast NMD occurs minimally if at all on CBC-bound transcripts, arguing against a CBC-mediated pioneer round of translation in yeast. The data taken together indicate that there are no direct consequences of a PTC within the yeast nucleus. PMID- 15547137 TI - Artificial tertiary motifs stabilize trans-cleaving hammerhead ribozymes under conditions of submillimolar divalent ions and high temperatures. AB - Tertiary stabilizing motifs (TSMs) between terminal loops or internal bulges facilitate folding of natural hammerhead ribozymes (hRz) under physiological conditions. However, both substrate and enzyme strands contribute nucleotides to the TSMs of trans-cleaving hRz, complicating the design of hRz that exploit TSMs to target specific mRNA. To overcome this limitation, we used SELEX to identify new, artificial TSMs that are less sensitive to sequence context. Nucleotides in loop II or in a bulge within the ribozyme strand of stem I were randomized, while the interaction partner was held constant. All nucleotides of the substrate pair with the ribozyme, minimizing their possible recruitment into the TSM, as such recruitment could constrain choice of candidate target sequences. Six cycles of selection identified cis-acting ribozymes that were active in 100 microM MgCl2. The selected motifs partially recapitulate TSMs found in natural hRz, suggesting that the natural motifs are close to optimal for their respective contexts. Ribozyme "RzB" showed enhanced thermal stability by retaining trans-cleavage activity at 80 degrees C in 10 mM MgCl2 and at 70 degrees C in 2 mM MgCl2. A variant of ribozyme "RzB" with a continuously paired stem 1 rapidly lost activity as temperature was increased. The selected motifs are modular, in that they permit trans-cleavage of several substrates in submillimolar MgCl2, including two substrates derived from the U5 genomic region of HIV-1. The new, artificial tertiary stabilized hRz are thus nearly independent of sequence context and enable for the first time the use of highly active hRz targeting almost any mRNA at physiologically relevant magnesium concentrations. PMID- 15547138 TI - Activation of the protein kinase PKR by short double-stranded RNAs with single stranded tails. AB - The human RNA-activated protein kinase PKR is an interferon-induced protein that is part of the innate immune response and inhibits viral replication. The action of PKR involves RNA-dependent autophosphorylation leading to inhibition of translation. PKR has an N-terminal dsRNA-binding domain that can interact non sequence specifically with long (>33 bp) stretches of dsRNA leading to activation. In addition, certain viral and cellular RNAs containing non-Watson Crick structures and multiple, shorter dsRNA sections can regulate PKR. In an effort to identify novel binders and possible activators of PKR, we carried out selections on a partially structured dsRNA library using truncated and full length versions of PKR. A library with 10(11) sequences was constructed and aptamers that bound to His6-tagged proteins were isolated. Characterization revealed a novel minimal RNA motif for activation of PKR with the following unified structural characteristics: a hairpin with a nonconserved imperfect 16-bp dsRNA stem flanked by 10-15-nt single-stranded tails, herein termed a "ss-dsRNA motif." Boundary experiments revealed that the single-stranded tails flanking the dsRNA core provide the critical determinant for activation. The ss-dsRNA motif occurs in a variety of cellular and viral RNAs, suggesting possible novel functions for PKR in nature. PMID- 15547139 TI - Mapping of genetic determinants of the sympathoneural response to stress. AB - Activation of the sympathoadrenal system (SAS, comprising the sympathetic nervous system and the adrenal medulla) in response to stressful stimuli is an important defense mechanism as well as a contributor to several cardiovascular diseases. There is variability in the SAS response to stress, although the extent to which this is genetically regulated is unclear. Some rodent models, including the hereditary hypertriglyceridemic (hHTg) rat, are hyperresponsive to stress. We investigated whether quantitative trait loci (QTLs) that affect sympathoadrenal response to stress could be identified. Second filial generation rats (n = 189) derived from a cross of the hHTg rat and the Brown Norway rat had plasma norepinephrine (NE) and epinephrine (Epi) levels, indices of activation of the sympathoneural and adrenal medulla components, respectively, measured in the resting state and in response to an immobilization stress. Responses were assessed early (20 min) and late (120 min) after the application of the stress. A genome scan was conducted using 153 microsatellite markers. Two QTLs (maximum peak LOD scores of 4.17 and 3.52, respectively) influencing both the early and late plasma NE response to stress were found on chromosome 10. Together, the QTLs accounted for approximately 20% of the total variation in both the early and late NE responses in the F(2) rats. Interestingly, the QTLs had no effect on plasma Epi response to stress. These findings provide evidence for a genetic determination of the response of a specific component of the SAS response to stress. Genetically determined variation in sympathetic nervous system response to stress may contribute to cardiovascular diseases. PMID- 15547140 TI - It takes time to make a pinky: unexpected insights into how SHH patterns vertebrate digits. AB - It is widely accepted that the diffusible Sonic Hedgehog (SHH) morphogen signal establishes a spatial gradient that patterns embryonic structures by long-range signaling. In response, cell fates are determined by linear thresholds according to the position of cells within the gradient field. Two recent studies of SHH signaling during vertebrate limb development challenge this spatial gradient model. They establish that a large fraction of limb bud cells patterned by SHH are descendants of cells that have previously expressed Shh. These cells are endowed with a kinetic memory that integrates their exposure to SHH rather than sensing their position in a SHH gradient. In addition, a fraction of cells changes their SHH responsiveness progressively during limb bud pattering, which is indicative of local nonlinear modulation of cell fate specification. PMID- 15547141 TI - AMPK activation is not critical in the regulation of muscle FA uptake and oxidation during low-intensity muscle contraction. AB - To determine the role of AMP-activated protein kinase (AMPK) activation on the regulation of fatty acid (FA) uptake and oxidation, we perfused rat hindquarters with 6 mM glucose, 10 microU/ml insulin, 550 microM palmitate, and [14C]palmitate during rest (R) or electrical stimulation (ES), inducing low-intensity (0.1 Hz) muscle contraction either with or without 2 mM 5-aminoimidazole-4-carboxamide-1 beta-D-ribofuranoside (AICAR). AICAR treatment significantly increased glucose and FA uptake during R (P < 0.05) but had no effect on either variable during ES (P > 0.05). AICAR treatment significantly increased total FA oxidation (P < 0.05) during both R (0.38 +/- 0.11 vs. 0.89 +/- 0.1 nmol x min(-1) x g(-1)) and ES (0.73 +/- 0.11 vs. 2.01 +/- 0.1 nmol x min(-1) x g(-1)), which was paralleled in both conditions by a significant increase and significant decrease in AMPK and acetyl-CoA carboxylase (ACC) activity, respectively (P < 0.05). Low-intensity muscle contraction increased glucose uptake, FA uptake, and total FA oxidation (P < 0.05) despite no change in AMPK (950.5 +/- 35.9 vs. 1,067.7 +/- 58.8 nmol x min(-1) x g(-1)) or ACC (51.2 +/- 6.7 vs. 55.7 +/- 2.0 nmol x min(-1) x g(-1)) activity from R to ES (P > 0.05). When contraction and AICAR treatment were combined, the AICAR-induced increase in AMPK activity (34%) did not account for the synergistic increase in FA oxidation (175%) observed under similar conditions. These results suggest that while AMPK-dependent mechanisms may regulate FA uptake and FA oxidation at rest, AMPK-independent mechanisms predominate during low-intensity muscle contraction. PMID- 15547142 TI - Involvement of calcium-sensing receptor in osteoblastic differentiation of mouse MC3T3-E1 cells. AB - We have previously shown that the extracellular calcium-sensing receptor (CaR) is expressed in various bone marrow-derived cell lines and plays an important role in stimulating their proliferation and chemotaxis. It has also been reported that the CaR modulates matrix production and mineralization in chondrogenic cells. However, it remains unclear whether the CaR plays any role in regulating osteoblast differentiation. In this study, we found that mineralization of the mouse osteoblastic MC3T3-E1 cells was increased when the cells were exposed to high calcium (2.8 and 3.8 mM) or a specific CaR activator, NPS-R467 (1 and 3 microM). Next, we stably transfected MC3T3-E1 cells with either a CaR antisense vector (AS clone) or a vector containing the inactivating R185Q variant of the CaR (DN clone) that has previously been shown to exert a dominant negative action. Alkaline phosphatase activities were decreased compared with controls in both the AS and DN clones. However, the levels of type I procollagen and osteopontin mRNA in the AS clone, as detected by Northern blotting, were almost the same as in the controls. On the other hand, the expression of osteocalcin, which is expressed at a later stage of osteoblastic differentiation, was significantly reduced in both the AS and DN clones. Mineralization was also decreased in both clones. In conclusion, this study showed that the abolition of CaR function results in diminishing alkaline phosphatase activity, osteocalcin expression, and mineralization in mouse osteoblastic cells. This suggests that the CaR may be involved in osteoblastic differentiation. PMID- 15547144 TI - Expanded newborn genetic testing urged. PMID- 15547145 TI - Is PSA testing still useful? PMID- 15547146 TI - New imaging approaches unveiled: techniques reveal clues to body's biochemistry. PMID- 15547154 TI - HAART and sexual risk behavior. PMID- 15547155 TI - HAART and sexual risk behavior. PMID- 15547156 TI - Isoflavones and postmenopausal women. PMID- 15547157 TI - Isoflavones and postmenopausal women. PMID- 15547158 TI - Isoflavones and postmenopausal women. PMID- 15547159 TI - Proteomic Profiling and Intra-amniotic Infection. PMID- 15547160 TI - Role of Toll-like receptor 4 in acute myocardial infarction and longevity. PMID- 15547161 TI - Regression of electrocardiographic left ventricular hypertrophy during antihypertensive treatment and the prediction of major cardiovascular events. AB - CONTEXT: Electrocardiographic left ventricular hypertrophy (LVH) is a strong predictor of cardiovascular (CV) morbidity and mortality. However, the predictive value of changes in the magnitude of electrocardiographic LVH criteria during antihypertensive therapy remains unclear. OBJECTIVE: To test the hypothesis that lesser severity of electrocardiographic LVH during antihypertensive treatment is associated with decreased CV morbidity and mortality, independent of blood pressure levels and reduction and treatment modality. DESIGN, SETTING, AND PARTICIPANTS: Double-blind, randomized, parallel-group study conducted in 1995 2001 among 9193 men and women with hypertension aged 55 through 80 years (mean, 67 years), with electrocardiographic LVH by Cornell voltage-duration product or Sokolow-Lyon voltage criteria and enrolled in the Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) study. INTERVENTIONS: Losartan- or atenolol-based treatment regimens, with follow-up assessments for at least 4 (mean, 4.8 [SD, 0.9]) years. MAIN OUTCOME MEASURE: Composite end point of CV death, myocardial infarction (MI), or stroke in relation to severity of electrocardiographic LVH determined at baseline and on subsequent electrocardiograms obtained at 1 or more annual revisits. RESULTS: Cardiovascular death, nonfatal MI, or stroke occurred in 1096 patients (11.9%). In Cox regression models controlling for treatment type, baseline Framingham risk score, baseline and in-treatment blood pressure, and severity of baseline electrocardiographic LVH by Cornell product and Sokolow-Lyon voltage, less-severe in-treatment LVH by Cornell product and Sokolow-Lyon voltage were associated with 14% and 17% lower rates, respectively, of the composite CV end point (adjusted hazard ratio [HR], 0.86; 95% confidence interval [CI], 0.82-0.90; P<.001 for every 1050-mm x ms [1-SD] decrease in Cornell product; and HR, 0.83; 95% CI, 0.78 0.88; P<.001 for every 10.5-mm [1-SD] decrease in Sokolow-Lyon voltage). In parallel analyses, lower Cornell product and Sokolow-Lyon voltage were each independently associated with lower risks of CV mortality (HR, 0.78; 95% CI, 0.73 0.83; P<.001; and HR, 0.80; 95% CI, 0.73-0.87; P<.001, respectively), MI (HR, 0.90; 95% CI, 0.82-0.98; P=.01; and HR, 0.90; 95% CI, 0.81-1.00; P = .04), and stroke (HR, 0.90; 95% CI, 0.84-0.96; P=.002; and HR, 0.81; 95% CI, 0.75-0.89; P<.001). CONCLUSIONS: Less-severe electrocardiographic LVH by Cornell product and Sokolow-Lyon voltage criteria during antihypertensive therapy is associated with lower likelihoods of CV morbidity and mortality, independent of blood pressure lowering and treatment modality in persons with essential hypertension. Antihypertensive therapy targeted at regression or prevention of electrocardiographic LVH may improve prognosis. PMID- 15547162 TI - Prognostic significance of left ventricular mass change during treatment of hypertension. AB - CONTEXT: Increased baseline left ventricular (LV) mass predicts cardiovascular (CV) complications of hypertension, but the relation between lower LV mass and outcome during treatment for hypertension is uncertain. OBJECTIVE: To determine whether reduction of LV mass during antihypertensive treatment modifies risk of major CV events independent of blood pressure change. DESIGN, SETTING, AND PARTICIPANTS: Prospective cohort substudy of the Losartan Intervention For Endpoint Reduction in Hypertension (LIFE) randomized clinical trial, conducted from 1995 to 2001. A total of 941 prospectively identified patients aged 55 to 80 years with essential hypertension and electrocardiographic LV hypertrophy had LV mass measured by echocardiography at enrollment in the LIFE trial and thereafter were followed up annually for a mean (SD) of 4.8 (1.0) years for CV events. MAIN OUTCOME MEASURES: Composite end point of CV death, fatal or nonfatal myocardial infarction, and fatal or nonfatal stroke. RESULTS: The composite end point occurred in 104 patients (11%). The multivariable Cox regression model showed a strong association between lower in-treatment LV mass index and reduced rate of the composite CV end point (hazard ratio [HR], 0.78 per 1-SD (25.3) decrease in LV mass index; 95% confidence interval [CI], 0.65-0.94; P = .009) over and above that predicted by reduction in blood pressure. There were parallel associations between lower in-treatment LV mass index and lower CV mortality (HR, 0.62; 95% CI, 0.47-0.82; P = .001), stroke (HR, 0.76; 95% CI, 0.60-0.96; P = .02), myocardial infarction (HR, 0.85; 95% CI, 0.62-1.17, P = .33), and all-cause mortality (HR, 0.72; 95% CI, 0.59-0.88, P = .002), independent of systolic blood pressure and assigned treatment. Results were confirmed in analyses adjusting for additional CV risk factors, electrocardiographic changes, or when only considering events after the first year of study treatment. CONCLUSION: In patients with essential hypertension and baseline electrocardiographic LV hypertrophy, lower LV mass during antihypertensive treatment is associated with lower rates of clinical end points, additional to effects of blood pressure lowering and treatment modality. PMID- 15547163 TI - Neurodevelopmental and growth impairment among extremely low-birth-weight infants with neonatal infection. AB - CONTEXT: Neonatal infections are frequent complications of extremely low-birth weight (ELBW) infants receiving intensive care. OBJECTIVE: To determine if neonatal infections in ELBW infants are associated with increased risks of adverse neurodevelopmental and growth sequelae in early childhood. DESIGN, SETTING, AND PARTICIPANTS: Infants weighing 401 to 1000 g at birth (born in 1993 2001) were enrolled in a prospectively collected very low-birth-weight registry at academic medical centers participating in the National Institute of Child Health and Human Development Neonatal Research Network. Neurodevelopmental and growth outcomes were assessed at a comprehensive follow-up visit at 18 to 22 months of corrected gestational age and compared by infection group. Eighty percent of survivors completed the follow-up visit and 6093 infants were studied. Registry data were used to classify infants by type of infection: uninfected (n = 2161), clinical infection alone (n = 1538), sepsis (n = 1922), sepsis and necrotizing enterocolitis (n = 279), or meningitis with or without sepsis (n = 193). MAIN OUTCOME MEASURES: Cognitive and neuromotor development, neurologic status, vision and hearing, and growth (weight, length, and head circumference) were assessed at follow-up. RESULTS: The majority of ELBW survivors (65%) had at least 1 infection during their hospitalization after birth. Compared with uninfected infants, those in each of the 4 infection groups were significantly more likely to have adverse neurodevelopmental outcomes at follow-up, including cerebral palsy (range of significant odds ratios [ORs], 1.4-1.7), low Bayley Scales of Infant Development II scores on the mental development index (ORs, 1.3 1.6) and psychomotor development index (ORs, 1.5-2.4), and vision impairment (ORs, 1.3-2.2). Infection in the neonatal period was also associated with impaired head growth, a known predictor of poor neurodevelopmental outcome. CONCLUSIONS: This large cohort study suggests that neonatal infections among ELBW infants are associated with poor neurodevelopmental and growth outcomes in early childhood. Additional studies are needed to elucidate the pathogenesis of brain injury in infants with infection so that novel interventions to improve these outcomes can be explored. PMID- 15547164 TI - Inpatient computer-based standing orders vs physician reminders to increase influenza and pneumococcal vaccination rates: a randomized trial. AB - CONTEXT: Computerized reminder systems increase influenza and pneumococcal vaccination rates, but computerized standing order systems have not been previously described or evaluated. OBJECTIVE: To determine the effects of computerized physician standing orders compared with physician reminders on inpatient vaccination rates. DESIGN, SETTING, AND PATIENTS: Randomized trial of 3777 general medicine patients discharged from 1 of 6 study wards during a 14 month period (November 1, 1998, through December 31, 1999) composed of 2 overlapping influenza seasons at an urban public teaching hospital. INTERVENTIONS: The hospital's computerized physician order entry system identified inpatients eligible for influenza and pneumococcal vaccination. For patients with standing orders, the system automatically produced vaccine orders directed to nurses at the time of patient discharge. For patients with reminders, the computer system provided reminders to physicians that included vaccine orders during routine order entry sessions. MAIN OUTCOME MEASURE: Vaccine administration. RESULTS: During the approximately 6 months of the influenza season, 50% of all hospitalized patients were identified as eligible for influenza vaccination. Twenty-two percent of patients hospitalized during the entire 14 months of the study were found eligible for pneumococcal vaccination. Patients with standing orders received an influenza vaccine significantly more often (42%) than those patients with reminders (30%) (P <.001). Patients with standing orders received a pneumococcal vaccine significantly more often (51%) than those with reminders (31%) (P <.001). CONCLUSIONS: Computerized standing orders were more effective than computerized reminders for increasing both influenza and pneumococcal vaccine administration. Our findings suggest that computerized standing orders should be used more widely for this purpose. PMID- 15547165 TI - Ozone and short-term mortality in 95 US urban communities, 1987-2000. AB - CONTEXT: Ozone has been associated with various adverse health effects, including increased rates of hospital admissions and exacerbation of respiratory illnesses. Although numerous time-series studies have estimated associations between day-to day variation in ozone levels and mortality counts, results have been inconclusive. OBJECTIVE: To investigate whether short-term (daily and weekly) exposure to ambient ozone is associated with mortality in the United States. DESIGN AND SETTING: Using analytical methods and databases developed for the National Morbidity, Mortality, and Air Pollution Study, we estimated a national average relative rate of mortality associated with short-term exposure to ambient ozone for 95 large US urban communities from 1987-2000. We used distributed-lag models for estimating community-specific relative rates of mortality adjusted for time-varying confounders (particulate matter, weather, seasonality, and long-term trends) and hierarchical models for combining relative rates across communities to estimate a national average relative rate, taking into account spatial heterogeneity. MAIN OUTCOME MEASURE: Daily counts of total non-injury-related mortality and cardiovascular and respiratory mortality in 95 large US communities during a 14-year period. RESULTS: A 10-ppb increase in the previous week's ozone was associated with a 0.52% increase in daily mortality (95% posterior interval [PI], 0.27%-0.77%) and a 0.64% increase in cardiovascular and respiratory mortality (95% PI, 0.31%-0.98%). Effect estimates for aggregate ozone during the previous week were larger than for models considering only a single day's exposure. Results were robust to adjustment for particulate matter, weather, seasonality, and long-term trends. CONCLUSIONS: These results indicate a statistically significant association between short-term changes in ozone and mortality on average for 95 large US urban communities, which include about 40% of the total US population. The findings indicate that this widespread pollutant adversely affects public health. PMID- 15547166 TI - Effects of systematic prone positioning in hypoxemic acute respiratory failure: a randomized controlled trial. AB - CONTEXT: A recent trial showed that placing patients with acute lung injury in the prone position did not increase survival; however, whether those results hold true for patients with hypoxemic acute respiratory failure (ARF) is unclear. OBJECTIVE: To determine whether prone positioning improves mortality in ARF patients. DESIGN, SETTING, AND PATIENTS: Prospective, unblinded, multicenter controlled trial of 791 ARF patients in 21 general intensive care units in France using concealed randomization conducted from December 14, 1998, through December 31, 2002. To be included, patients had to be at least 18 years, hemodynamically stable, receiving mechanical ventilation, and intubated and had to have a partial pressure of arterial oxygen (PaO2) to fraction of inspired oxygen (FIO2) ratio of 300 or less and no contraindications to lying prone. INTERVENTIONS: Patients were randomly assigned to prone position placement (n = 413), applied as early as possible for at least 8 hours per day on standard beds, or to supine position placement (n = 378). MAIN OUTCOME MEASURES: The primary end point was 28-day mortality; secondary end points were 90-day mortality, duration of mechanical ventilation, incidence of ventilator-associated pneumonia (VAP), and oxygenation. RESULTS: The 2 groups were comparable at randomization. The 28-day mortality rate was 32.4% for the prone group and 31.5% for the supine group (relative risk [RR], 0.97; 95% confidence interval [CI], 0.79-1.19; P = .77). Ninety-day mortality for the prone group was 43.3% vs 42.2% for the supine group (RR, 0.98; 95% CI, 0.84 1.13; P = .74). The mean (SD) duration of mechanical ventilation was 13.7 (7.8) days for the prone group vs 14.1 (8.6) days for the supine group (P = .93) and the VAP incidence was 1.66 vs 2.14 episodes per 100-patients days of intubation, respectively (P = .045). The PaO2/FIO2 ratio was significantly higher in the prone group during the 28-day follow-up. However, pressure sores, selective intubation, and endotracheal tube obstruction incidences were higher in the prone group. CONCLUSIONS: This trial demonstrated no beneficial outcomes and some safety concerns associated with prone positioning. For patients with hypoxemic ARF, prone position placement may lower the incidence of VAP. PMID- 15547167 TI - Management of fibromyalgia syndrome. AB - CONTEXT: The optimal management of fibromyalgia syndrome (FMS) is unclear and comprehensive evidence-based guidelines have not been reported. OBJECTIVE: To provide up-to-date evidence-based guidelines for the optimal treatment of FMS. DATA SOURCES, SELECTION, AND EXTRACTION: A search of all human trials (randomized controlled trials and meta-analyses of randomized controlled trials) of FMS was made using Cochrane Collaboration Reviews (1993-2004), MEDLINE (1966-2004), CINAHL (1982-2004), EMBASE (1988-2004), PubMed (1966-2004), Healthstar (1975 2000), Current Contents (2000-2004), Web of Science (1980-2004), PsychInfo (1887 2004), and Science Citation Indexes (1996-2004). The literature review was performed by an interdisciplinary panel, composed of 13 experts in various pain management disciplines, selected by the American Pain Society (APS), and supplemented by selected literature reviews by APS staff members and the Utah Drug Information Service. A total of 505 articles were reviewed. DATA SYNTHESIS: There are major limitations to the FMS literature, with many treatment trials compromised by short duration and lack of masking. There are no medical therapies that have been specifically approved by the US Food and Drug Administration for management of FMS. Nonetheless, current evidence suggests efficacy of low-dose tricyclic antidepressants, cardiovascular exercise, cognitive behavioral therapy, and patient education. A number of other commonly used FMS therapies, such as trigger point injections, have not been adequately evaluated. CONCLUSIONS: Despite the chronicity and complexity of FMS, there are pharmacological and nonpharmacological interventions available that have clinical benefit. Based on current evidence, a stepwise program emphasizing education, certain medications, exercise, cognitive therapy, or all 4 should be recommended. PMID- 15547168 TI - Left ventricular hypertrophy: the next treatable, silent killer? PMID- 15547169 TI - Developmental vulnerability and resilience in extremely preterm infants. PMID- 15547170 TI - JAMA patient page. Left ventricular hypertrophy. PMID- 15547171 TI - Can selenium prevent colorectal cancer? A signpost from epidemiology. PMID- 15547172 TI - Is there a new role for induction chemotherapy in the treatment of head and neck cancer? PMID- 15547173 TI - Full-body CT screening: preventing or producing cancer? PMID- 15547174 TI - Silencing the critics: new studies move closer to answering epigenetic questions. PMID- 15547175 TI - NIH clarifies new model organisms sharing policy. PMID- 15547176 TI - Federal ruling requires million-year guarantee of safety at Yucca Mountain nuclear waste site. PMID- 15547177 TI - Stat bite: Frequency of breast cancer screening by health plan, 2000-2003. PMID- 15547178 TI - The role of BRCA1 in the cellular response to chemotherapy. AB - Germline mutations of the BRCA1 gene account for approximately 5% of breast and ovarian cancer cases, and lower than normal BRCA1 expression or function may be an important contributing factor in sporadic cancers. The major role of BRCA1 is to respond to DNA damage by participating in cellular pathways for DNA repair, mRNA transcription, cell cycle regulation, and protein ubiquitination. Because most chemotherapeutic agents function by directly or indirectly damaging DNA, the role of BRCA1 as a regulator of chemotherapy-induced DNA damage has been the subject of an increasing number of investigations. We review published preclinical and clinical evidence that the level of BRCA1 function in an individual patient's tumor can guide the choice of chemotherapeutic agents for breast and ovarian cancer. We conclude that a loss of BRCA1 function is associated with sensitivity to DNA-damaging chemotherapy and may also be associated with resistance to spindle poisons. We recommend that prospective clinical studies investigating the role of BRCA1 in the response to chemotherapy be conducted. PMID- 15547179 TI - Selenium and colorectal adenoma: results of a pooled analysis. AB - BACKGROUND: Secondary analyses of data from a large randomized clinical trial have suggested that intake of the trace element selenium reduces risk of colorectal neoplasia, but epidemiologic studies have not shown a consistent protective association. METHODS: We conducted a combined analysis of data from three randomized trials--the Wheat Bran Fiber Trial, the Polyp Prevention Trial, and the Polyp Prevention Study--which tested the effects of various nutritional interventions for colorectal adenoma prevention among participants who recently had an adenoma removed during colonoscopy. Selenium concentrations were measured from blood specimens from a total of 1763 trial participants, and quartiles of baseline selenium were established from the pooled data. To estimate the association between baseline selenium and colorectal adenoma risk, odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using logistic regression modeling. All statistical tests were two-sided. RESULTS: Individual study results among participants whose blood selenium concentrations were in the highest versus the lowest quartile varied in magnitude (Polyp Prevention Trial: OR = 0.67, 95% CI = 0.43 to 1.05; P(trend) = .21; Wheat Bran Fiber Trial: OR = 0.66, 95% CI = 0.40 to 1.10; P(trend) = .13, and Polyp Prevention Study: OR = 0.57, 95% CI = 0.34 to 0.95, P(trend) = .04). Analyses of the pooled data showed that individuals whose blood selenium values were in the highest quartile (median = 150 ng/mL) had statistically significantly lower odds of developing a new adenoma compared with those in the lowest quartile (OR = 0.66, 95% CI = 0.50 to 0.87; P(trend) = .006). CONCLUSIONS: The inverse association between higher blood selenium concentration and adenoma risk supports previous findings indicating that higher selenium status may be related to decreased risk of colorectal cancer. PMID- 15547180 TI - Clinically relevant pneumonitis after sequential paclitaxel-based chemotherapy and radiotherapy in breast cancer patients. AB - BACKGROUND: Taxane-based chemotherapy has been associated with an increased risk of radiation pneumonitis in patients with breast cancer. To obtain additional information about this association, we investigated the association between paclitaxel chemotherapy and radiation pneumonitis in patients participating in a phase III randomized study. METHODS: Five hundred and twenty-four breast cancer patients were prospectively and randomly assigned to receive either four cycles of paclitaxel followed by four cycles of 5-fluorouracil, doxorubicin, cyclophosphamide (FAC) or eight cycles of FAC. One hundred and eighty-nine of these patients (100 in the paclitaxel-FAC group and 89 in the FAC group) subsequently underwent radiation therapy in our institution and had medical records available to review for pulmonary symptoms. In addition, a radiologist who was unaware of the type of treatment scored chest x-ray changes after radiation treatment. Crude rates of radiation pneumonitis were compared with chi square or Fisher's exact test, and actuarial rates were assessed with Kaplan Meier and log-rank tests. All statistical tests were two-sided. RESULTS: No difference in the rate of clinically relevant radiation pneumonitis was observed between the two groups (5.0% in the paclitaxel-FAC group versus 4.5% in the FAC group; difference = 0.5%, 95% CI = -6.6% to 5.5%; P = 1.00). Oral steroids for pneumonitis were taken by two patients in the paclitaxel-FAC group but by none in the FAC group, and no patient was hospitalized for or died of radiation pneumonitis. The paclitaxel-FAC group (39.3%) had a higher rate of radiographic changes after irradiation than the FAC group (23.7%; difference = 15.6%, 95% CI = -0.11% to 28.8%; P = .034). CONCLUSION: Patients with breast cancer treated with sequential paclitaxel, FAC, and radiation therapy appeared to have a very low rate of clinically relevant radiation pneumonitis that was no different from that of patients treated with FAC alone. PMID- 15547181 TI - Phase III randomized trial of docetaxel-carboplatin versus paclitaxel-carboplatin as first-line chemotherapy for ovarian carcinoma. AB - BACKGROUND: Chemotherapy with a platinum agent and a taxane (paclitaxel) is considered the standard of care for treatment of ovarian carcinoma. We compared the combination of docetaxel-carboplatin with the combination of paclitaxel carboplatin as first-line chemotherapy for stage Ic-IV epithelial ovarian or primary peritoneal cancer. METHODS: We randomly assigned 1077 patients to receive docetaxel at 75 mg/m2 of body surface area (1-hour intravenous infusion) or paclitaxel at 175 mg/m2 (3-hour intravenous infusion). Both treatments then were followed by carboplatin to an area under the plasma concentration-time curve of 5. The treatments were repeated every 3 weeks for six cycles; in responding patients, an additional three cycles of single-agent carboplatin was permitted. Survival curves were calculated by the Kaplan-Meier method, and hazard ratios were estimated with the Cox proportional hazards model. All statistical tests were two-sided. RESULTS: After a median follow-up of 23 months, both groups had similar progression-free survival (medians of 15.0 months for docetaxel carboplatin and 14.8 months for paclitaxel-carboplatin; hazard ratio [HR] docetaxel-paclitaxel = 0.97, 95% confidence interval [CI] = 0.83 to 1.13; P = .707), overall survival rates at 2 years (64.2% and 68.9%, respectively; HR = 1.13, 95% CI = 0.92 to 1.39; P = .238), and objective tumor (58.7% and 59.5%, respectively; difference between docetaxel and paclitaxel = -0.8%, 95% CI = -8.6% to 7.1%; P = .868) and CA-125 (75.8% and 76.8%, respectively; difference docetaxel-paclitaxel = -1.0%, 95% CI = -7.2% to 5.1%; P = .794) response rates. However, docetaxel-carboplatin was associated with substantially less overall and grade 2 or higher neurotoxicity than paclitaxel-carboplatin (grade > or =2 neurosensory toxicity in 11% versus 30%, difference = 19%, 95% CI = 15% to 24%; P<.001; grade > or =2 neuromotor toxicity in 3% versus 7%, difference = 4%, 95% CI = 1% to 7%; P<.001). Treatment with docetaxel-carboplatin was associated with statistically significantly more grade 3-4 neutropenia (94% versus 84%, difference = 11%, 95% CI = 7% to 14%; P<.001) and neutropenic complications than treatment with paclitaxel-carboplatin, although myelosuppression did not influence dose delivery or patient safety. Global quality of life was similar in both arms, but substantive differences in many symptom scores favored docetaxel. CONCLUSIONS: Docetaxel-carboplatin appears to be similar to paclitaxel carboplatin in terms of progression-free survival and response, although longer follow-up is required for a definitive statement on survival. Thus, docetaxel carboplatin represents an alternative first-line chemotherapy regimen for patients with newly diagnosed ovarian cancer. PMID- 15547182 TI - Reactivation of latent Epstein-Barr virus by methotrexate: a potential contributor to methotrexate-associated lymphomas. AB - BACKGROUND: Patients with rheumatoid arthritis or polymyositis treated with methotrexate (MTX) develop Epstein-Barr virus (EBV)-positive lymphomas more frequently than patients treated with other, equally immunosuppressive regimens. Here we determined whether MTX, in contrast to other commonly used medications for rheumatoid arthritis or polymyositis, is unique in its ability to induce the release of infectious EBV from latently infected cells. METHODS: The effect of MTX and other immunosuppressant drugs on EBV replication in vitro was assessed using latently infected EBV-positive lymphoblastoid and gastric carcinoma cell lines. Inhibitors of signal transduction pathways were used to define requirements for induction of lytic infection. Drug effects on transcription of the two EBV immediate-early promoters (BRLF1 and BZLF1) and on promoter constructs lacking cis-acting sequences required for activation by other effectors was examined using reporter gene assays. EBV viral load in rheumatoid arthritis and polymyositis patients receiving MTX was compared with that in patients receiving other immunosuppressive medications. Statistical tests were two-sided. RESULTS: MTX activated the release of infectious EBV from latently infected cell lines in vitro, and MTX treatment was associated with activation of the two viral immediate-early promoters in reporter gene assays. Induction of lytic EBV infection by MTX required the p38 MAP kinase, PI3 kinase, and MEK pathways and specific cis-acting motifs in the two viral immediate-early promoters. Patients treated with MTX-containing regimens had statistically significantly higher mean EBV loads in their blood than patients treated with immunosuppressing regimens that did not include MTX (40 EBV copies per 10(6) cellular genomes versus 5.1 copies; geometric mean fold difference in copies = 10.8, 95%, confidence interval = 3.0 to 38; P = .011). CONCLUSION: MTX may promote EBV-positive lymphomas in rheumatoid arthritis and polymyositis patients by its immunosuppressive properties as well as by reactivating latent EBV. PMID- 15547183 TI - Effect of proton pump inhibitor pretreatment on resistance of solid tumors to cytotoxic drugs. AB - BACKGROUND: Resistance to antitumor agents is a major cause of treatment failure in patients with cancer. Some mechanisms of tumor resistance to cytotoxic drugs may involve increased acidification of extracellular compartments. We investigated whether proton pump inhibitors (PPIs), currently used in the anti acid treatment of peptic disease, could inhibit the acidification of the tumor microenvironment and increase the sensitivity of tumor cells to cytotoxic agents. METHODS: We pretreated cell lines derived from human melanomas, adenocarcinomas, and lymphomas with the PPIs omeprazole, esomeprazole, or pantoprazole and tested their response to cytotoxic drugs in cell death assays. We also evaluated extracellular and intracellular pH and vacuolar-H+-ATPase (V-H+-ATPase) expression, distribution, and activity in PPI-pretreated cells by using western blot analyses, immunocytochemistry, laser scanning confocal analysis, and bioluminescence assays. Finally, we evaluated human melanoma growth and cisplatin sensitivity with or without omeprazole pretreatment in xenografted SCID/SCID mice. RESULTS: PPI pretreatment sensitized tumor cell lines to the effects of cisplatin, 5-fluorouracil, and vinblastine, with an IC50 value reduction up to 2 logs. PPI pretreatment was associated with the inhibition of V-H+-ATPase activity and increases in both extracellular pH and the pH of lysosomal organelles. PPI pretreatment induced a marked increase in the cytoplasmic retention of the cytotoxic drugs, with clear targeting to the nucleus in the case of doxorubicin. In in vivo experiments, oral pretreatment with omeprazole was able to induce sensitivity of human solid tumors to cisplatin. CONCLUSION: Our results open new possibilities for the treatment of drug-resistant tumors through combination strategies based on the use of well-tolerated pH modulators such as PPIs. PMID- 15547184 TI - Randomized phase III trial of neoadjuvant chemotherapy in head and neck cancer: 10-year follow-up. AB - In 1986, we initiated a multicenter, randomized trial to compare induction chemotherapy with cisplatin and 5-fluorouracil followed by locoregional treatment (surgery and radiotherapy or radiotherapy alone) with locoregional treatment alone in patients with head and neck squamous cell carcinoma. Here we report the long-term results of the trial. A total of 237 patients with nonmetastatic stage III or IV head and neck carcinoma were randomly assigned to receive four cycles of neoadjuvant chemotherapy followed by locoregional treatment (group A) or locoregional treatment alone (group B). Among all patients, overall survival at 5 and 10 years was 23% (95% confidence interval [CI] = 15.3% to 30.9%) and 19% (95% CI = 11.6% to 26.4%), respectively, for those in group A and 16% (95% CI = 9.6% to 23.4%) and 9% (95% CI = 3.5% to 14.7%), respectively, for those in group B (P = .13). Among operable patients, we observed no difference between group A and group B in overall survival at 5 and 10 years (group A, 31% [95% CI = 14.9% to 47.3%] and 22.7% [95% CI = 7.1% to 38.3%], respectively; group B, 43.3% [95% CI = 25.6% to 61.0%] and 14.2% [95% CI = 0.1% to 28.3%], respectively; P = .73). Among inoperable patients, overall survival at 5 and 10 years was 21% (95% CI = 12.3% to 30.1%) and 16% (95% CI = 7.7% to 23.9%), respectively, for group A and 8% (95% CI = 1.5% to 12.3%) and 6% (95% CI = 0.1% to 9.1%), respectively, for group B (log-rank P = .04). Four cycles of neoadjuvant chemotherapy is a promising approach for treating patients with inoperable advanced head and neck cancer but not for treating patients with operable disease. PMID- 15547185 TI - Expression of Pirh2, a newly identified ubiquitin protein ligase, in lung cancer. AB - Maintenance of p53 function is important for normal cell growth and development, and loss of p53 function contributes directly to malignant tumor development. The recently discovered Pirh2 protein is an ubiquitin-protein ligase that negatively regulates p53 through activity by targeting it for degradation. To determine how Pirh2 may mediate lung tumorigenesis, we evaluated Pirh2 expression in human and mouse lung tumor samples and paired normal lung tissues using immunoblot analysis and immunohistochemistry. Pirh2 protein expression was higher in 27 (84%) of 32 human lung neoplasms than in matched normal lung tissue and in 14 of 15 mouse lung tumors evaluated. In addition, p53 protein was more ubiquitinated in the mouse lung tumors than in normal tissue, and overall p53 expression was lower than that in normal tissue. These results are consistent with the hypothesis that increased Pirh2 expression affects lung tumorigenesis by reducing p53 activity. To our knowledge, this is the first description of altered Pirh2 expression in human and mouse tumors. PMID- 15547186 TI - Re: Assessing the probability that a positive report is false: an approach for molecular epidemiology studies. PMID- 15547187 TI - Development of c-Kit-expressing small-cell lung cancer in a chronic myeloid leukemia patient during imatinib treatment. PMID- 15547188 TI - Randomized controlled trials. PMID- 15547189 TI - Peer review at the American Journal of Roentgenology: how reviewer and manuscript characteristics affected editorial decisions on 196 major papers. AB - OBJECTIVE: The objective of this study was to examine the relative influence of manuscript characteristics and peer-reviewer attributes in the assessment of manuscripts. MATERIALS AND METHODS: Over a 6-month period, all major papers submitted to the American Journal of Roentgenology (AJR) were entered into a database that recorded manuscript characteristics, demographic profiles of reviewers, and the disposition of the manuscript. Manuscript characteristics included reviewer ratings on five scales (rhetoric, structure, science, import, and overall recommendation); the subspecialty class of the paper; the primary imaging technique; and the country of origin. Demographic profiles of the reviewers included age, sex, subspecialty, years of reviewing, academic rank, and practice type. Statistical analysis included correlation analysis, ordinal logistic regression, and analysis of variance. RESULTS: A total of 445 reviews of 196 manuscripts were the work of 335 reviewers. Of the 196 submitted manuscripts, 20 (10.2%) were accepted, 106 (54.1%) were rejected, and 70 (35.7%) were rejected with the opportunity to resubmit. Regarding manuscript characteristics, we found that the country of origin, score on the science scale, and score on the import scale were statistically significant variables for predicting the final disposition of a manuscript. Of the reviewer attributes, we found a statistically significant association between greater reviewer age and also higher academic rank with lower scores on the import scale. Reviewer concordance was higher for structure, science, and overall scores than on the rhetoric and import scores. Greater variability in the overall scoring of papers could be attributed to the reviewer than the manuscript, but both factors combined explain only 23% of the total variability. CONCLUSION: At the AJR, manuscript acceptance was most strongly associated with reviewer scoring of the science and import of a major paper and also with the country of origin. Reviewers who were older and of higher academic rank tended to discount the importance of manuscripts. PMID- 15547190 TI - Dual-phase 3D MDCT angiography for evaluation of the liver before hepatic resection. AB - OBJECTIVE: We sought to evaluate the accuracy of dual-phase MDCT angiography for assessing the liver before hepatic resection and to compare 2D and 3D images for quality and arterial branch visualization. MATERIALS AND METHODS: Sixty-three patients with colorectal metastases (n = 30), hepatocellular carcinomas (n = 13), giant hemangiomas (n = 5), and other lesions (n = 15) underwent dual-phase MDCT using either a LightSpeed QX/i 4-MDCT (n = 31) or LightSpeed QX/i Ultra 8-MDCT (n = 32) scanner. Contrast material (150 mL of Isovue 370 [iopamidol]) was injected at a rate of 5 mL/sec. The arterial phase images were rendered on a workstation to obtain 3D MDCT angiograms that were assessed by two reviewers who were blinded to the surgical findings. Arterial anatomy was categorized according to the Michels classification. The reviewers assessed the 2D and 3D images for quality, arterial branch visualization, and differences between the 4- and 8-MDCT images. In the 43 patients who underwent resection, imaging findings were correlated with intraoperative findings. RESULTS: The anatomy of hepatic arteries in the 63 patients was classified as follows: Michels type I, 51 patients (80.9%); type III, four patients (6.3%); type V, five patients (7.9%); and types VII, VIII, and IX, one patient (1.6%) each. In 40 (93%) of 43 patients, the surgical findings concurred with MDCT findings. Three discrepancies were due to failure to identify small accessory left hepatic arteries. Branch visualization and image quality of the 2D images were superior to those of the 3D images. No significant difference was found between the 4- and 8-MDCT images in branch visualization and image quality. CONCLUSION: Three-dimensional MDCT angiography is accurate for classification of hepatic arterial anatomy before hepatic resection. Although 2D data sets show small arteries to better advantage than 3D MDCT angiograms, the 3D MDCT angiograms provide a useful overview of hepatic anatomy. PMID- 15547191 TI - Early biliary complications of laparoscopic cholecystectomy: evaluation on T2 weighted MR cholangiography in conjunction with mangafodipir trisodium-enhanced 3D T1-weighted MR cholangiography. AB - OBJECTIVE: Our aim was to assess preliminary experience with combined conventional T2-weighted and mangafodipir trisodium (MnDPDP)-enhanced T1-weighted MR cholangiography in evaluating early biliary complications of laparoscopic cholecystectomy. SUBJECTS AND METHODS: Conventional heavily T2-weighted MR cholangiography with MnDPDP-enhanced T1-weighted MR cholangiography and ERCP were performed in seven patients with high clinical suspicion of biliary complications after laparoscopic cholecystectomy. The final diagnoses of complications were classified according to the presence and degree of bile duct injury, bile leakage, and retained stones. RESULTS: The diagnoses on MR cholangiography were as follows: complete transection and occlusion of the common bile duct with bile leakage (n = 3), partial strictures of the common bile duct with bile leakage (n = 1), cystic duct leakage (n = 1), partial ligation of an aberrant right hepatic duct (n = 1), and hemorrhage without biliary complication (n = 1). The final diagnoses at surgery (n = 2) and ERCP (n = 5) were as follows: complete transection and occlusion of the common bile duct with bile leakage (n = 2), partial strictures of the common bile duct with bile leakage (n = 2), cystic duct leakage (n = 1), partial ligation of an aberrant right hepatic duct (n = 1), and hemorrhage without biliary complication (n = 1). MR cholangiography accurately yielded the same findings as the final diagnoses, except in one case with partial stricture of the bile duct with bile leakage (overdiagnosed as complete occlusion on MR cholangiography). CONCLUSION: Combined conventional T2-weighted and MnDPDP enhanced T1-weighted MR cholangiography may eliminate the use of other studies for the imaging of biliary complications after cholecystectomy if this preliminary data can be verified in a larger study. PMID- 15547192 TI - Value of MR cholangiography in patients with iatrogenic bile duct injury after cholecystectomy. AB - OBJECTIVE: Our aim was to determine the diagnostic role of MR cholangiography in the evaluation of iatrogenic bile duct injuries after cholecystectomy. SUBJECTS AND METHODS: Nineteen patients (14 women and five men; mean age, 47 years; age range, 24-75 years) with suspected bile duct injury as a result of laparoscopic cholecystectomy (17 patients) and open cholecystectomy (two patients) underwent MR cholangiography. MR images were evaluated for bile duct discontinuity, presence or absence of biliary dilation, stricture, excision injury, free fluid, and collections. Bile duct excision and stricture were classified according to the Bismuth classification. Final diagnosis was made on the basis of findings at surgery in 15 patients, on percutaneous transhepatic cholangiography (PTC) in one patient, and on endoscopic retrograde cholangiography (ERC) and at clinical follow-up until hospital discharge in the remaining three patients. RESULTS: In 16 patients, injury of the bile duct was observed. Two patients had Bismuth type I injury; one patient, type II injury; 11 patients, type III injury; and one patient each, type IV and V injuries. Three patients showed findings suggestive of leakage from the cystic duct remnant, which were confirmed on ERC. CONCLUSION: MR cholangiography is an accurate diagnostic technique in the identification of postoperative bile duct injuries. This technique allows exploration above and below the level of obstruction, a resource provided by neither ERC nor PTC, and allows the accurate classification of these injuries, which is essential for treatment planning. PMID- 15547193 TI - CT of benign hypervascular liver nodules in autoimmune hepatitis. AB - OBJECTIVE: The purpose of this report is to describe the frequency and histopathologic basis of benign hypervascular liver nodules seen on CT in patients with autoimmune hepatitis. CONCLUSION: Benign hypervascular liver nodules may be seen on CT in patients with cirrhosis due to autoimmune hepatitis and may represent large regenerative nodules. This phenomenon is important to recognize because of the potential for confusion with hepatocellular carcinoma. PMID- 15547194 TI - Does variant hepatic artery anatomy in a liver transplant recipient increase the risk of hepatic artery complications after transplantation? AB - OBJECTIVE: Our aim was to determine whether variant hepatic artery anatomy in a liver transplant recipient increases the risk of hepatic artery complications after liver transplantation. MATERIALS AND METHODS: The study group consisted of 84 patients who underwent gadolinium-enhanced 3D MR angiography before orthotopic liver transplantation in which a branch patch arterial anastomosis at the gastroduodenal takeoff was used. MR angiography studies were retrospectively reviewed and assessed for the presence and type of variant hepatic artery anatomy. The diameter of the distal common hepatic artery was measured. The incidence of posttransplantation hepatic artery stenosis or thrombosis was assessed. RESULTS: Seven (8.3%) of the 84 patients developed hepatic artery complications after transplantation. Of the 24 patients with variant hepatic artery anatomy, five (20.8%) had posttransplantation/ hepatic artery complications. In contrast, only two (3.3%) of the 60 patients with classic hepatic artery anatomy had complications. The higher complication rate in patients with variant hepatic artery anatomy was statistically significant (p < 0.05). The odds ratio was 7.6 (95% confidence interval, 1.4-42.6). The diameter of the distal common hepatic artery was smaller in patients with variant hepatic artery anatomy compared with those with classic hepatic artery anatomy (range, 4.3-7.1 mm [mean, 5.8 mm] vs 4.0-8.9 mm [mean 6.3 mm], p < 0.05), and it was also smaller in patients who had posttransplantation hepatic artery complications compared with those who had no complications (range, 4.2-6.3 mm [mean, 5.2 mm] vs 4.0-8.9 mm, [mean, 6.2 mm], p < 0.01). CONCLUSION: Variant hepatic artery anatomy in a liver transplant recipient increased the risk of hepatic artery complications after transplantation. The smaller caliber of the native common hepatic artery may contribute to the higher risk. PMID- 15547195 TI - Expression of vascular endothelial growth factor in hepatocellular carcinoma and the surrounding liver: correlation with angiographically assisted CT. AB - OBJECTIVE: The purpose of our study was to assess the correlation between the intensity and characteristics of contrast enhancement on angiographically assisted CT and the intensity of vascular endothelial growth factor (VEGF) expression in hepatocellular carcinoma (HCC) and in the surrounding nontumorous liver. MATERIALS AND METHODS: The intensity of VEGF expression in HCC and in the surrounding liver was expressed as a VEGF expression index by Western blot analysis in 20 surgical specimens resected in 20 patients between March 2000 and August 2002. Findings on CT during arterial portography (n = 20) and CT hepatic arteriography (n = 17) were retrospectively evaluated to determine contrast enhancement indexes and the enhancement characteristics of HCCs and of the surrounding liver. Contrast enhancement indexes and VEGF expression indexes were correlated using a simple regression test, and enhancement characteristics and VEGF expression indexes were correlated using the Spearman's rank correlation test. RESULTS: On CT hepatic arteriography, the contrast enhancement indexes of HCCs showed moderate inverse correlation with the VEGF expression indexes of HCCs (r = -0.57, p = 0.017) and high inverse correlation with the differences between the VEGF expression indexes of HCCs and those of livers (difference in the VEGF expression index, -0.80; p = 0.0001). The contrast enhancement index of the liver showed marginal moderate direct correlation with the VEGF expression index of the liver (0.44, p = 0.076) and high inverse correlation with the difference in the VEGF expression index (-0.71, p = 0.0013). On CT during arterial portography, the contrast enhancement indexes of HCCs showed moderate inverse correlation with the difference in the VEGF expression index (-0.51, p = 0.023). The qualitative degree of heterogeneity of hepatic artery enhancement in HCC on CT hepatic arteriography showed moderate direct correlation with the VEGF expression indexes of HCCs (0.55, p = 0.033) and high direct correlation with the difference in the VEGF expression indexes (0.73, p = 0.004). CONCLUSION: Our results indicated that the intensity and heterogeneity of hepatic artery enhancement of HCCs on CT hepatic arteriography correlated with the degree of VEGF expression in HCCs. PMID- 15547196 TI - CT and MRI of cirrhosis and its mimics. PMID- 15547197 TI - Volumetric contrast imaging in bile duct sonography: technology and early clinical experience. PMID- 15547198 TI - Hepatic steatosis and pancreatitis associated with the use of stavudine in a patient with HIV infection. PMID- 15547199 TI - Metastatic cerebellar medulloblastoma in the liver mimicking a complicated cyst: sonographic and MDCT findings. PMID- 15547200 TI - Changes in bile ducts after radiofrequency ablation of hepatocellular carcinoma: frequency and clinical significance. AB - OBJECTIVE: The purpose of our study was to determine the frequency of bile duct changes after radiofrequency ablation for hepatocellular carcinoma and to evaluate their clinical significance. MATERIALS AND METHODS: Between April 1999 and August 2003, 389 patients with 521 hepatocellular carcinomas underwent a total of 571 sessions of radiofrequency ablation. The maximum dimension of the tumors measured on sonography was 2.4 +/- 0.9 cm (mean +/- SD) (range, 0.5-5.0 cm). The frequency and type of bile duct changes resulting from radiofrequency ablation, the time interval between radiofrequency ablation and the first appearance of bile duct changes, and the serial changes at follow-up CT were analyzed. Complications related to bile duct changes were also evaluated by reviewing medical records and CT scans. RESULTS: Bile duct changes occurred in 69 (12%) of 571 sessions and 66 (17%) of 389 patients. Bile duct changes seen on CT included mild dilatation of upstream bile ducts surrounding the ablation zone in 57 patients (82.6%), biloma in the ablation zone in four patients (5.8%), and both in eight patients (11.6%). The mean time interval between radiofrequency ablation and the initial appearance of bile duct change was 1.6 months (range, immediate-9 months). Most (87%) of the 69 patients with bile duct changes showed no progression on follow-up CT, and only nine (13%) had slight progression. All patients but one, in whom cholangitis developed, had no major complications requiring specific treatment during the follow-up period. CONCLUSION: Although bile duct changes were frequent after the radiofrequency ablation of hepatocellular carcinoma, most were of no clinical significance, and major complications requiring additional treatment were rare. PMID- 15547201 TI - CT evaluation of the response of gastrointestinal stromal tumors after imatinib mesylate treatment: a quantitative analysis correlated with FDG PET findings. AB - OBJECTIVE: We correlated changes in tumor density on CT with changes in glucose metabolism, or the maximum standardized uptake value (SUV(max)), on FDG PET and sought to develop CT imaging criteria that can be used to objectively evaluate tumor response in patients with metastatic gastrointestinal stromal tumors (GISTs) who undergo treatment with imatinib mesylate. MATERIALS AND METHODS: Using the criteria established by the Response Evaluation Criteria in Solid Tumors (RECIST) group, we selected 173 tumors (in 36 patients) for study. Tumor size and density were determined objectively, and overall tumor response (OTR) was evaluated subjectively on CT images. The changes in these parameters before and after treatment were correlated with changes in SUV(max). RESULTS: Significant decreases were seen in both tumor density (mean, 12.3 H [16.5%]; p < 0.0001) and SUV(max) (mean, 3.43 [64.9%]; p < 0.0001). OTR evaluated subjectively, correlated well with changes in SUV(max) (p < 0.0001). No statistically significant association was found between changes in tumor density and changes in SUV(max) (p = 0.3088), but 70% (14/20) of the patients with tumors that showed response on FDG PET exhibited at least a partial response by a change in tumor density. Tumor size was found to have decreased significantly 2 months after treatment (p = 0.0070). However, in 75% of the patients, the disease was stable according to the traditional tumor response criteria of RECIST. CONCLUSION: FDG PET is sensitive and specific for evaluating tumor response but cannot be used in patients whose baseline FDG PET results are negative for tumors. Although subjective evaluation was a better indicator of treatment response than was tumor density alone, the tumor density measurement is a good indicator and provides a reliable quantitative means of monitoring the tumor. RECIST, using only tumor size, was unreliable for monitoring GISTs during the early stage of imatinib mesylate treatment. PMID- 15547202 TI - Gastrointestinal stromal tumors in patients with neurofibromatosis: imaging features with clinicopathologic correlation. AB - OBJECTIVE: The purpose of this study was to evaluate the clinical, pathologic, and imaging features of gastrointestinal stromal tumors that occur in patients with neurofibromatosis. CONCLUSION: Gastrointestinal stromal tumors that occur in patients with neurofibromatosis commonly originate from the proximal small intestine and are often multiple. The cross-sectional imaging appearance of gastrointestinal stromal tumors that occur in patients with neurofibromatosis is similar to that of gastrointestinal stromal tumors that occur in the general population. PMID- 15547203 TI - MRI of pancreatitis and its complications: part 1, acute pancreatitis. PMID- 15547204 TI - MRI of pancreatitis and its complications: part 2, chronic pancreatitis. PMID- 15547205 TI - CT features of the accessory spleen. AB - OBJECTIVE: The purpose of this study was to describe the prevalence and CT features of the accessory spleen. CONCLUSION: Accessory spleens are present in 16% of patients undergoing contrast-enhanced abdominal CT. Typically, accessory spleens appear on CT scans as well-marginated, round masses that are smaller than 2 cm and enhance homogeneously on contrast-enhanced images. When accessory spleens are smaller than 1 cm, their attenuation may be lower than that of the spleen because of partial volume effects. Their most frequent location (22%) is posteromedial to the spleen; anterolateral to the upper pole of the left kidney; and lateral, posterior, and superior to the tail of the pancreas. Familiarity with these characteristic features may differentiate them from other pathologic findings in the upper abdomen. PMID- 15547206 TI - CT findings of an abdominal cocoon. PMID- 15547207 TI - The effect of diagnostic confidence on the probability of optical colonoscopic confirmation of potential polyps detected on CT colonography: prospective assessment in 1,339 asymptomatic adults. AB - OBJECTIVE: We sought to evaluate the effect of interpreter confidence on the likelihood that a lesion detected on CT colonography (CTC) will correspond to a matched polyp seen on optical colonoscopy. SUBJECTS AND METHODS: Same-day CTC and optical colonoscopy were performed on 1,339 asymptomatic adults. A standard matching algorithm for polyp size and location was used. For each potential polyp detected on CTC, the level of diagnostic confidence was prospectively rated on a 3-point scale (1, least certain; 2, intermediate; and 3, most certain). RESULTS: For CTC-detected lesions 6 mm or larger, diagnostic confidence levels of 1, 2, and 3 corresponded to matched polyps on optical colonoscopy in 33.3% (45/135), 50.0% (103/206), and 66.8% (157/235) of cases, respectively (p < 0.01). Similar trends were present for categories of lesions that measured 6-7 mm, 8-9 mm, and 10 mm or larger, rising to a match rate of 82.1% (55/67) for lesions 10 mm or larger that were diagnosed with a level-3 confidence rating. The likelihood that a matched polyp was adenomatous increased with greater levels of diagnostic confidence. Of note, level-3 confidence for lesions measuring 8-9 mm on CTC more often yielded a matching neoplasm on optical colonoscopy than level-1 or level-2 confidence for lesions measuring 10 mm or larger (60.3% [35/58] vs 20.8% [10/48]; p < 0.0001). CONCLUSION: Greater diagnostic confidence for an individual lesion detected on CTC correlates with a significantly increased likelihood that a matching polyp will be found on optical colonoscopy and that this matched polyp will be neoplastic. Although polyp size represents the primary criterion for CTC screening algorithms, this data could help guide the decision to opt for noninvasive CTC surveillance versus optical colonoscopy for polypectomy. PMID- 15547208 TI - Cecal pneumatosis in patients with obstructive colon cancer: correlation of CT findings with bowel viability. AB - OBJECTIVE: The purpose of our study was to reassess CT findings of cecal pneumatosis in patients with acute large-bowel obstruction due to colon cancer to determine whether this condition indicates transmural necrosis versus viable bowel and also whether other CT findings could be used to identify patients with transmural necrosis. CONCLUSION: CT findings of cecal pneumatosis do not always indicate transmural infarction in patients with acute large-bowel obstruction due to colon cancer. Cecal pneumatosis may be related to viable bowel when it displays a bubblelike pattern or when it is not associated with other findings of ischemia. PMID- 15547209 TI - Contrast-induced nephropathy. PMID- 15547210 TI - Antegrade MDCT pyelography for the evaluation of patients with obstructed urinary tract. AB - OBJECTIVE: The purpose of this study was to design an alternative imaging technique to CT urography in patients with high-grade urinary tract obstruction, with or without impaired renal function, that uses the superior resolution of MDCT and avoids IV contrast material administration. CONCLUSION: Antegrade MDCT pyelography is an alternative imaging technique to CT urography in patients with high-grade urinary tract obstruction with or without impaired renal function. It enables accurate diagnosis of the level of obstruction, as well as its etiology, including nephroureterolithiasis, urothelial tumors, primary congenital megaureter, uretero-pelvic junction stenosis, ureteral edema, ureteral stricture, retroperitoneal fibrosis, and pelvic lymphadenopathy. PMID- 15547211 TI - Differential diagnosis of perinephric masses on CT and MRI. PMID- 15547212 TI - Malacoplakia presenting as a solitary renal mass. PMID- 15547213 TI - Transrectal color Doppler sonographically guided compression to treat active extravasation after transrectal prostate biopsy. PMID- 15547214 TI - Necrotizing fasciitis caused by xanthogranulomatous and emphysematous pyelonephritis: importance of the inferior lumbar triangle pathway. PMID- 15547215 TI - Adrenal metastasis from ovarian carcinoma. PMID- 15547216 TI - MRI guidance of focused ultrasound therapy of uterine fibroids: early results. AB - OBJECTIVE: The purpose of this study was to explore our hypothesis that MRI guided focused ultrasound therapy for the treatment of uterine fibroids will lead to a significant reduction in symptoms and improvement in quality of life. We describe focused ultrasound therapy applications and the method for monitoring the thermal energy deposited in the fibroids, including the MRI parameters required, in a prospective review of 108 treatments. MATERIALS AND METHODS: Patients presenting with symptomatic uterine fibroids who attained a minimal symptom severity score and who would otherwise have been offered a hysterectomy were recruited. Thermal lesions were created within target fibroids using an MRI guided focused ultrasound therapy system. The developing lesion was monitored using real-time MR thermometry, which was used to assess treatment outcome in real time to change treatment parameters and achieve the desired outcome. Fibroid volume, fibroid symptoms, and quality-of-life scores were measured before treatment and 6 months after treatment. Adverse events were actively monitored and recorded. RESULTS: In this study, 79.3% of women who had been treated reported a significant improvement in their uterine fibroid symptoms on follow-up health-related quality-of-life questionnaires, which supports our hypothesis. The mean reduction in fibroid volume at 6 months was 13.5%, but nonenhancing volume (mean, 51 cm(3)) remained within the treated fibroid at 6 months. CONCLUSION: This early description of MRI-guided focused ultrasound therapy treatment of fibroids includes follow-up data and shows that, although the volume reduction is moderate, it correlates with treatment volume and the symptomatic response to this treatment is encouraging. PMID- 15547217 TI - CT features of serous surface papillary carcinoma of the ovary. AB - OBJECTIVE: The purpose of this study was to determine the CT appearance of serous surface papillary carcinoma of the ovary. MATERIALS AND METHODS: CT scans of 17 patients with histologically proven serous surface papillary carcinoma of the ovary were retrospectively reviewed. We evaluated the ovary size, omental or mesenteric involvement (none, mild, or severe), ascites (none, small, moderate, or large), and peritoneal thickening (none, smooth, or nodular). We also noted the presence of a cul-de-sac mass, lymphadenopathy, and calcification within the peritoneal mass. The preoperative serum level of cancer antigen (CA)-125 was assessed in all patients. RESULTS: The diameter of ovaries was 3 cm or smaller in 14 patients (82%). All patients had omental or mesenteric involvement by the tumor (mild, n = 2; severe, n = 15) and ascites (small, n = 1; moderate, n = 3; large, n = 13). Peritoneal thickening (smooth, n = 5; nodular, n = 10) was noted in 15 patients (88%) and a cul-de-sac mass in 10 (59%). Lymphadenopathy was noted in five patients (29%) and calcification within the peritoneal mass in one (6%). Serum CA-125 level was elevated in all patients (168-63,300 U/mL). CONCLUSION: Serous surface papillary carcinoma of the ovary should be suggested as a diagnosis in patients who have peritoneal carcinomatosis, relatively normal-sized ovaries, and a highly elevated serum CA-125 level. PMID- 15547218 TI - Enhanced fertility after diagnostic hysterosalpingography using oil-based contrast agents may be attributable to immunomodulation. PMID- 15547219 TI - Enhanced fertility after diagnostic hysterosalpingography may be a myth. PMID- 15547220 TI - MR arthrography of the hip: diagnostic performance of a dedicated water excitation 3D double-echo steady-state sequence to detect cartilage lesions. AB - OBJECTIVE: The objective of our study was to compare the diagnostic performance of a dedicated cartilage MR sequence (water-excitation 3D double-echo steady state) with a standard MR sequence (T1-weighted spin-echo) in detecting articular cartilage lesions of the hip after intraarticular injection of gadopentetate dimeglumine. MATERIALS AND METHODS: In 50 MR arthrograms of the hip joint obtained in 47 consecutive patients, a sagittal 3D double-echo steady-state sequence (TR/TE, 24/6.5; flip angle, 25 degrees ) was compared with a sagittal T1 weighted spin-echo sequence (350/14). Two musculoskeletal radiologists independently evaluated articular cartilage. Sensitivity and specificity for detecting cartilage defects were calculated for those hips that underwent open surgery (n = 21). Lesion conspicuity was retrospectively reviewed and graded between 1 (not visible) and 5 (well defined). RESULTS: At surgery, a total of 26 lesions of the acetabular (n = 20) and femoral (n = 6) cartilage were found. For the 3D double-echo steady-state and T1-weighted spin-echo sequences, sensitivities and specificities for cartilage lesion detection were 58% and 88% and 81% and 81% for reviewer 1 and 62% and 94% and 62% and 100% for reviewer 2, respectively. Lesion conspicuity was significantly superior (p = 0.036) for the 3D double-echo steady-state sequence (mean grade, 3.4) compared with the T1 weighted spin-echo sequence (mean grade, 3.0). The kappa value was fair for the 3D double-echo steady-state sequence (kappa = 0.40) and moderate for the T1 weighted spin-echo sequence (kappa = 0.55). CONCLUSION: The 3D double-echo steady state sequence optimized for cartilage imaging improves lesion conspicuity but does not improve diagnostic performance. PMID- 15547221 TI - MRI of adamantinoma of long bones in correlation with histopathology. AB - OBJECTIVE: The purposes of this retrospective study were to assess specific MRI features of adamantinoma, including classic adamantinoma and its osteofibrous dysplasia-like variant, and to assess the role of adamantinoma in surgical planning. MATERIALS AND METHODS: MR images of 22 patients with histologically proven adamantinoma, subtyped according to defined criteria, were analyzed, with emphasis on morphologic features, signal intensities, and enhancement parameters. Intra- and extraosseous tumor extent was determined. In all patients, examination of the corresponding resected specimens was performed with regard to tumor extent and presence of multicentricity. Moreover, radiographs were reviewed, and radiographic features derived from the literature were determined. RESULTS: All tumors were primarily localized in the tibia diaphysis (including one patient with additional lesions in the fibula), most frequently in the anterior cortical bone (19/22) with extension toward the bone marrow in 12 patients. We distinguished two morphologic patterns: a solitary lobulated focus versus a pattern of multiple small nodules in one or more foci. Separated tumor foci, defined as foci of high signal intensity on either T2-weighted images or T1 weighted contrast-enhanced images, interspersed with normal-appearing cortical or spongious bone were seen in six patients. All tumors showed intense and homogeneous static enhancement, but there was no uniform dynamic enhancement pattern. No relationship between MRI features and histologic subtype of adamantinoma was found. CONCLUSION: Some uniform MRI characteristics, along with those of radiography, may contribute to the diagnosis of adamantinoma; however, these are not related to the histologic subtype. MRI is pivotal for precise locoregional staging, especially for depiction of distant cortical foci, soft tissue, and intramedullary extension and thus is useful for determining tumor free margins and strategies for reconstructive surgery. PMID- 15547222 TI - Sublabral sulcus at the posteroinferior acetabulum: a potential pitfall in MR arthrography diagnosis of acetabular labral tears. AB - OBJECTIVE: Our study correlated findings on hip MRI and MR arthrography with hip arthroscopy to assess the location, prevalence, and potential pitfall of a normal acetabular sublabral sulcus. MATERIALS AND METHODS: We retrospectively collected 58 hip MRI studies along with surgical reports in 58 patients who underwent hip arthroscopy over a 5-year period. Intraoperative photography (n = 23), radiography (n = 56), unenhanced MRI (n = 13), and MR arthrography (n = 54) studies were available for review. Two radiologists described hip anatomy on radiology studies with agreement by consensus. RESULTS: A normal posteroinferior sublabral groove was confirmed on available arthroscopy photographs in four (17.4%) of 23 hips. In each of these four patients, the anatomic sublabral groove correlated with apparent partial labral detachment on MR arthrography. On review of all studies, 13 hips (22.4%) without a posterior labral tear at surgery had imaging findings of a sublabral sulcus. The sulcus was not associated with acetabular dysplasia, which was radiographically noted in 12 cases (21.4%). Preoperatively, the sulcus was misdiagnosed as a tear in two cases. Labral tears were anterior or anterosuperior in 51 patients. CONCLUSION: A posteroinferior sublabral groove is a relatively common normal anatomic hip variation. If not recognized as normal, the sulcus may serve as a diagnostic pitfall on MR arthrography. Its location is distinct from most labral tears. We did not discover a sublabral sulcus at the anterior or anterosuperior acetabulum, the most common sites of labral injury. PMID- 15547223 TI - Validation of dental panoramic radiography measures for identifying postmenopausal women with spinal osteoporosis. AB - OBJECTIVE: Measurements of mandibular inferior cortical shape and width detected on dental panoramic radiographs may be a useful screening tool for spinal osteoporosis in postmenopausal women. The purposes of this study were to clarify whether these measures are validated compared with simple screening tools based on questionnaires, such as the osteoporosis self-assessment tool (OST) and whether these measures can be used in postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. SUBJECTS AND METHODS: We calculated the diagnostic performances of panoramic measurements and the OST for identifying women with spinal osteoporosis in both 159 healthy postmenopausal and 157 postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. Spinal osteoporosis was defined as a bone mineral density T score of -2.5 or less at the lumbar spine. Cortical shape and width were evaluated on dental panoramic radiographs. Receiver operating characteristic curve analyses were used to determine the optimal cutoff thresholds for cortical width and the OST in healthy postmenopausal women. RESULTS: The sensitivity and specificity, respectively, for identifying women with spinal osteoporosis were 89.5% and 33.9% for cortical width, 86.8% and 57.8% for the OST, and 86.8% and 63.6% for cortical shape in healthy postmenopausal women. Sensitivity and specificity, respectively, were 92.5% and 35.0% for cortical width, 72.5% and 58.1% for the OST, and 80.0% and 64.1% for cortical shape in postmenopausal women with histories of hysterectomy, oophorectomy, or estrogen use. CONCLUSION: Dentists may be able to refer postmenopausal women with suspected spinal osteoporosis for bone densitometry on the basis of dental panoramic radiographs with diagnostic performance similar to that of osteoporosis screening tools based on questionnaires. PMID- 15547224 TI - Bone marrow fat and bone mineral density on proton MR spectroscopy and dual energy X-ray absorptiometry: their ratio as a new indicator of bone weakening. AB - OBJECTIVE: Bone weakening can be affected by agents other than bone mineral density (BMD). Increased bone marrow fat may have a direct link to bone loss. This pilot study analyzes the relationship between bone marrow fat and BMD in subjects with normal and structurally weakened vertebrae. SUBJECTS AND METHODS: Twenty-six subjects underwent both dual-energy X-ray absorptiometry and proton MR spectroscopy of 71 lumbar vertebrae. Fifteen subjects had normal-appearing vertebrae on MRI, and 11 had signs of bone weakening. RESULTS: We found that high bone marrow fat did not consistently equate with low BMD. Bone marrow fat can indicate bone weakening nearly as well as BMD, but neither parameter alone is suitable to be used independently as an indicator. The bone marrow fat/BMD ratio showed significant diagnostic power to detect bone weakening, even in this relatively small subject sample. CONCLUSION: An inverse relationship between bone marrow fat and BMD could not be confirmed. Bone marrow fat can be used to diagnose reduced bone strength nearly as well as BMD. The bone marrow fat/BMD ratio is a significant diagnostic indicator of bone weakening. PMID- 15547225 TI - Collagenous fibroma (desmoplastic fibroblastoma) of the shoulder. PMID- 15547226 TI - Type 1 primary hyperoxaluria in pediatric patients: renal sonographic patterns. AB - OBJECTIVE: Our aim was to review the sonographic features of type I primary hyperoxaluria in children and to correlate the sonographic patterns with the clinical development of end-stage renal disease (ESRD). MATERIALS AND METHODS: We performed a retrospective analysis of the clinical and imaging files of 13 patients with type I primary hyperoxaluria who were treated in one institution and of the sonographic patterns and the clinical follow-up reports. RESULTS: We encountered the following two sonographic patterns: medullary nephrocalcinosis in eight patients and cortical nephrocalcinosis in five patients. The sonographic appearance of cortical nephrocalcinosis is quite specific: a hyperechoic peripheral renal cortex with acoustic shadowing behind it. Medullary nephrocalcinosis is less specific because there are many other causes of hyperechoic pyramids. All patients with medullary nephrocalcinosis developed lithiasis during the course of the disease. All patients with cortical nephrocalcinosis but only two of eight with medullary nephrocalcinosis developed ESRD. CONCLUSION: Sonography can be used differentiate the two patterns of type 1 primary hyperoxaluria. The cortical nephrocalcinosis type carries a higher risk of developing ESRD. PMID- 15547227 TI - Low-dose MDCT and virtual bronchoscopy in pediatric patients with foreign body aspiration. AB - OBJECTIVE: The purpose of this study was to investigate the potential use of low tube-current MDCT virtual bronchoscopy for the evaluation of children with suspected foreign body aspiration. SUBJECTS AND METHODS: Low-tube-current MDCT was performed in 23 patients (10 girls, 13 boys) with a mean age of 3.3 years (9 months-13 years) with suspicion of foreign body aspiration. Chest radiographs were obtained before CT was performed. MDCT was performed using 25- to 50-mA tube currents. MDCT virtual bronchoscopy images were obtained. Neither sedation nor IV contrast medium was used during CT scanning. All patients underwent endoscopic evaluation within 24 hr after MDCT was performed. MDCT virtual bronchoscopy findings were retrospectively compared with the results of rigid bronchoscopy. RESULTS: The mean tube current was 35 mA (range, 25-50 mA). Imaging quality was excellent in nine studies (39%), good in 12 studies (52%), and poor in two studies (9%). Motion artifacts were present on several slices in five examinations. In 15 patients, all foreign bodies detected by conventional bronchoscopy were also revealed on MDCT virtual bronchoscopy. The foreign body was in the right main bronchus in six patients, in the bronchus intermedius in one patient, and in the left main bronchus in eight patients. No discordance was found between the two techniques. MDCT revealed hyperaeration of the ipsilateral lung in five patients, atelectasis in five patients, infiltration in three patients, and infiltration and bronchiectasis in two patients; it showed infiltration in four patients and atelectasis in one of eight patients without a foreign body detected. There were no abnormal findings in three patients. CONCLUSION: Evaluation of foreign body aspiration of the airway in children can be accomplished by using a low-tube-current MDCT protocol. It may be useful both in showing the exact location of a foreign body before bronchoscopy and in ruling out a foreign body in patients with a low level of suspicion and normal or nonspecific findings on chest radiography. PMID- 15547228 TI - MRI appearance of accessory breast tissue: a diagnostic consideration for an axillary mass in a peripubertal or pubertal girl. AB - OBJECTIVE: The purpose of this study was to describe the MRI appearance of accessory breast tissue that should be considered a diagnostic possibility in peripubertal or pubertal girls who present with an axillary mass along the course of the primitive milk streak. CONCLUSION: The MRI appearance of accessory breast tissue is of a mass discontinuous with-but with signal intensity and contrast enhancement characteristics similar to-normal breast parenchyma. PMID- 15547229 TI - Congenital intrahepatic portosystemic venous shunt in an infant with down syndrome. PMID- 15547230 TI - Assessment of cerebral microembolism during percutaneous radiofrequency ablation of lung tumors using diffusion-weighted imaging. AB - OBJECTIVE: It is well known that radiofrequency ablation generates microbubbles in the liver. We hypothesized that microbubbles generated during percutaneous radiofrequency ablation of lung tumors flow into the pulmonary veins and are distributed to the systemic arteries, as with radiofrequency ablation of liver tumors. To assess the risk of cerebral infarction during radiofrequency ablation of lung tumors, we performed diffusion-weighted imaging and, if possible, monitored microemboli in the carotid artery during radiofrequency ablation. SUBJECTS AND METHODS: We prospectively studied 20 patients (19 men and one woman) who underwent radiofrequency ablation of lung tumors. Pre- and postoperative MRI examinations were performed in all 20 patients, and during 17 radiofrequency ablation sessions, sonography was used to monitor whether microemboli were generated. RESULTS: Radiofrequency ablation was technically feasible for the treatment of selected pulmonary tumors. Microemboli, which were believed to represent microbubbles, were seen on sonography during three of the 17 radiofrequency ablation sessions. They were rarely observed when a lung tumor was small, the treatment session was brief, and the radiofrequency emission power was low. No new area of abnormal intensity was seen on postoperative MRI in all 20 patients. Although the microemboli were observed, MRI could not confirm infarction. CONCLUSION: We concluded that cerebral infarction as a result of microbubbles generated during radiofrequency ablation of lung tumors has a low possibility of becoming a clinical problem. PMID- 15547231 TI - Changes in magnetization transfer MRI correlate with spreading depression-induced astroglial reactivity and increased protein expression in mice. AB - OBJECTIVE: Gliosis refers to a range of glial cell transformations that vary according to specific brain pathologic states. Disease, however, is not a prerequisite for gliosis because glial reactivity may also be seen in regions of increased physiologic activity. Our study tests the hypothesis that high-field strength magnetization transfer MRI is a sensitive technique for detecting transient glial reactivity after experimental spreading depression, a relatively benign perturbation unaccompanied by cell injury. MATERIALS AND METHODS: Unilateral neocortical spreading depression was elicited in mouse cerebral hemispheres and confirmed by transcranial blood flow and extracellular potential measurements. After 3 days, mice were imaged at 4 T using magnetization transfer techniques. Astroglial reactivity was determined immunohistochemically, and protein expression in control and experimental hemispheres was compared using proteomic techniques. RESULTS: Sixteen ([mean +/- SD] +/- 3) spreading depressions (n = 10) were recorded in experimental hemispheres. Spreading depression was never observed in control hemispheres. At 3 days, an 8% decrease (p < 0.05, n = 4) in magnetization transfer signal intensity was measured in experimental hemispheres, which was associated with a 37% increase (p < 0.001, n = 4) in the intensity of glial fibrillary acidic protein staining. Proteomic analysis performed 3 days after the induction of spreading depression showed upregulation of at least 56 proteins, including extracellular and intracellular elements. CONCLUSION: Magnetization transfer at 4.0-T MRI is a sensitive method for detecting glial reactivity and changes in protein expression not associated with cell injury. These results suggest magnetization transfer MRI techniques may have potential for detecting glial reactivity in physiologic processes such as learning and in early disease states. PMID- 15547232 TI - Differentiation of cavernous hemangioma from schwannoma of the orbit: a dynamic MRI study. AB - OBJECTIVE: The purpose of this study was to determine the capability of dynamic contrast MRI to differentiate hemangioma from schwannoma of the orbit. MATERIALS AND METHODS: Sixteen patients (three males and 13 females; mean age, 39 +/- 17.3 [SD] years; age range, 10-71 years) with unilateral orbital tumors, including eight cavernous hemangiomas and eight schwannomas, were examined. In addition to conventional MRI, we performed a dynamic contrast study (fast spin-echo sequence, 20-sec interval) after bolus administration of the contrast material (gadopentetate dimeglumine, 0.1 mmol/kg). We evaluated the features of the contrast enhancement spread pattern and the tumors' time-intensity curves. RESULTS: In the early phase, all the hemangiomas started the enhancement from one point or portion, although all the schwannomas started the enhancement from a wide area. The difference in the contrast-enhancement spread pattern features between the two types of tumors was statistically significant (p < 0.0001). The gradient of the time-intensity curve did not show a significant difference. CONCLUSION: Hemangioma and schwannoma of the orbit can be differentiated by the contrast-enhancement spread pattern on dynamic MRI. PMID- 15547233 TI - Contrast-enhanced high-resolution MRI of invasive breast cancer: correlation with histopathologic subtypes. AB - OBJECTIVE: We sought to determine whether contrast-enhanced MRI could aid in the identification of the histopathologic subtypes of invasive ductal carcinoma. MATERIALS AND METHODS: We evaluated the contrast-enhanced MR images obtained in 62 women with invasive ductal carcinoma of no special type. The presence or absence of three distinct MRI findings-linear enhancement, a serrated border, and delayed rim enhancement-was evaluated. Classification and regression tree analyses were performed to construct the most efficient algorithm for predicting histopathologic subtype on the basis of dynamic MRI features. RESULTS: Histopathologic subtypes of the invasive ductal carcinomas were scirrhous carcinoma in 22 patients, solid tubular carcinoma in 14, and papillotubular carcinoma in 26. A lesion with a serrated border was observed in 28 (45.2%) of the 62 patients. Delayed rim enhancement was seen in 23 (37.1%) and linear enhancement in 20 (32.3%). Scirrhous carcinomas were closely associated with a serrated border (20/22 or 90.9%, p < 0.0001). Delayed rim enhancement was frequently observed in solid tubular carcinomas (12/14 or 85.7%, p < 0.0001) but was not typically seen in scirrhous carcinomas (1/22 or 4.5%, p < 0.0001). Linear enhancement showed relatively high prevalence in papillotubular carcinomas (13/26 or 50%) and low prevalence in solid tubular carcinomas (1/22 or 7%, p < 0.02). Histopathologic subtypes of invasive breast carcinoma of no special type could be correctly identified in 47 (75.8%) of 62 lesions using the diagnostic algorithm generated by the classification and regression tree analyses. CONCLUSION: MRI features showed a close relationship with histopathologic subtypes of invasive ductal carcinoma of no special type. Contrast-enhanced MRI can be a noninvasive diagnostic tool for histopathologic subtypes of invasive breast cancer. PMID- 15547234 TI - Mammographic appearance of lymphedema in a TRAM-reconstructed breast. PMID- 15547235 TI - Aortic valve calcification as a marker for aortic stenosis severity: assessment on 16-MDCT. AB - OBJECTIVE: The degree of valvular calcification in patients with aortic stenosis was determined with retrospectively ECG-gated 16-MDCT and correlated with the severity of stenosis assessed at cardiac catheterization. SUBJECTS AND METHODS: We conducted a prospective study of 72 patients (38 men and 34 women; mean age +/ SD, 69.5 +/- 8.8 years) with aortic stenosis who underwent 16-MDCT and cardiac catheterization. Aortic valve calcification was assessed using the aortic Agatston score, aortic mass score, and aortic volume score. Severity of aortic stenosis was classified at cardiac catheterization. Aortic valve area and peak-to peak and mean transvalvular gradients were correlated with the degree of calcification determined on MDCT. RESULTS: All measured aortic valve calcification scores were significantly higher in patients with severe aortic stenosis (n = 46) than in patients with moderate (n = 15) or mild (n = 11, p < 0.001) aortic stenosis. Aortic valve calcification scores were inversely related to aortic valve area (r = -0.67, p < 0.001 for aortic mass score) and correlated significantly with peak-to-peak (r = 0.70, p < 0.001) and mean transvalvular (r = 0.72, p < 0.001) gradients. No correlation between the aortic valve calcification and the total coronary calcium scores was observed. CONCLUSION: Aortic valve calcification assessed on 16-MDCT is associated with severity of aortic stenosis. Thus, aortic valve calcification scores should be calculated routinely in all patients undergoing MDCT for assessment of coronary calcification. High aortic valve calcification scores indicate possibly severe aortic stenosis and should prompt a further functional evaluation. PMID- 15547236 TI - Accuracy of CT in the diagnosis of pulmonary embolism: a systematic literature review. AB - OBJECTIVE: We sought to summarize systematically the published evidence describing the accuracy of contrast-enhanced helical CT for diagnosing pulmonary embolism. MATERIALS AND METHODS: We selected all systematic reviews published before December 2003 that evaluated the accuracy of CT angiography for the diagnosis of pulmonary embolism. We also selected all prospective studies from the same time period in the primary literature in which all subjects underwent both CT and conventional angiography, the latter being considered the reference standard. Articles were identified through a computerized MEDLINE search and by other means. The quality and content of each article were evaluated independently by pairs of researchers. RESULTS: Six systematic reviews and eight primary studies were selected. The combined sensitivities of CT for detecting pulmonary embolism ranged from 66% to 93% across the systematic reviews and the combined specificities ranged from 89% to 97%. Only one of the reviews reported a combined sensitivity of greater than 90%. Among the eight primary studies, the sensitivities ranged from 45% to 100% and specificities ranged from 78% to 100%. Only three of the eight primary studies reported a sensitivity greater than 90%. None of the primary studies used scanners with four or more detectors. CONCLUSION: A systematic literature review revealed a wide range of reported sensitivities, only a minority of which exceeded 90%. Pooled estimates of sensitivity and specificity reported by systematic literature reviews should be interpreted with caution because of potential selection bias and heterogeneity in the reviewed studies. Accuracy studies of recent generations of MDCT scanners are not yet available despite the current dissemination of this technology. PMID- 15547237 TI - Safety and feasibility of using a central venous catheter for rapid contrast injection rates. AB - OBJECTIVE: Our aim was to determine the safety and feasibility of using a central venous catheter for rapid contrast injections during CT. MATERIALS AND METHODS: An in vitro experiment was performed using a 7-French Arrow-Howes multilumen central venous catheter. Each catheter port was tested by varying contrast agent flow rates delivered by a power injector. Contrast media specifications were kept similar to routine clinical practice. The in vivo experiment included 104 cases in which rapid contrast injections, 3.0-5.0 mL/sec, were delivered through a central venous catheter for dynamic CT examinations. Patient monitoring for early complications of contrast extravasation, cardiac arrhythmia, and allergic reactions was performed. Contrast injections were monitored for pressure limitation, automatic flow-rate adjustment, and catheter injury. Chart review was performed for delayed complications of mediastinal hematoma, infection, or catheter malfunction. RESULTS: During the in vitro experiment, all desired flow rates, 3.0-9.9 mL/sec, could be delivered through the central venous catheter with no catheter injury. No immediate or early patient or catheter complications were observed during the in vivo experiment. Follow-up evaluation revealed that 18 blood cultures and one catheter culture were positive for bacterial growth. In a subgroup of 43 patients, five contrast injections were pressure-limited by the power injector, and only one had the flow rate automatically adjusted to 3.6 mL/sec from 4.0 mL/sec. CONCLUSION: Rapid contrast injection rates, at 3.0-5.0 mL/sec, through the Arrow-Howes multilumen central venous catheter are feasible and safe in the clinical setting. However, a strict protocol should be followed to avoid possible serious complications. PMID- 15547238 TI - Primary posterior mediastinal seminoma. PMID- 15547239 TI - Acute myocardial infarction as the first manifestation of left atrial myxoma. PMID- 15547240 TI - ECG-gated MDCT of left-to-right shunt flow through a ventricular septal defect. PMID- 15547241 TI - Beeper bump. PMID- 15547242 TI - MR angiography of subclavian steal syndrome: pitfalls and solutions. PMID- 15547243 TI - Metastatic colorectal carcinoma: an unusual cause of calcified pulmonary metastases. PMID- 15547244 TI - MRI of angiolipoma of the breast in Turner's syndrome. PMID- 15547245 TI - Mesenteric gastrointestinal stromal tumor in a patient with neurofibromatosis. PMID- 15547246 TI - Isoform-specific regulation of adenylyl cyclase function by disruption of membrane trafficking. AB - Oligomerization plays an important role in endoplasmic reticulum processing and membrane insertion (and ultimately in regulation of function) of a number of transmembrane spanning proteins. Furthermore, it is known that adenylyl cyclases (ACs), critical regulators of cellular functions, associate into higher order (dimeric) forms. However, the importance of these higher order aggregates in regulating adenylyl cyclase activity or trafficking to the cell membrane is unclear. Therefore, we examined the potential role of oligomerization in the membrane trafficking of adenylyl cyclase. For this purpose, the ability of full length adenylyl cyclase and various truncation mutants to self-assemble and to be targeted to the cell membrane was assessed. A truncation mutant comprised of the initial six transmembrane spanning domains and half of the C1 catalytic domain coimmunoprecipitated with full-length AC VI. Using both biotinylation assays and assessment of enzyme distribution using sucrose density gradients, we demonstrate that expression of this mutant in human embryonic kidney 293 cells impaired the ability of AC VI to traffic to the plasma membrane. Furthermore, mutant expression resulted in a significant reduction in adenylyl cyclase activity. The decrease in AC VI membrane expression was not caused by alterations in enzyme transcription. The effect of the mutant was specific for the AC V and VI isoforms and expression of the transmembrane M1 domain but not the C1a domain was required for the mutant to affect adenylyl cyclase activity. In aggregate, these data suggest that alterations in the ability of adenylyl cyclases to form higher order forms regulate both enzyme trafficking and enzyme activity. PMID- 15547247 TI - Nuclear localization of NADPH:cytochrome c (P450) reductase enhances the cytotoxicity of mitomycin C to Chinese hamster ovary cells. AB - Overexpression of endoplasmic reticulum-localized NADPH: cytochrome c (P450) reductase (NPR) in Chinese hamster ovary cells increases the hypoxic/aerobic differential toxicity of the mitomycins. Because considerable evidence indicates that DNA cross-links are the major cytotoxic lesions generated by the mitomycins, we proposed that bioactivation of the mitomycins in the nucleus close to the DNA target would influence the cytotoxicity of these drugs. The simian virus 40 large T antigen nuclear localization signal was fused to the amino-terminal end of a human NPR protein that lacked its membrane anchor sequence. Immunofluorescent imaging of transfected cell lines expressing the fusion protein confirmed the nuclear location of the enzyme. Regardless of the oxygenation state of the cell, mitomycin C (MC) cytotoxicity was enhanced in cells with overexpressed NPR localized to the nuclear compartment compared with cells overexpressing an endoplasmic reticulum localized enzyme. Enhanced cytotoxicity in cells treated under hypoxic conditions correlated with increases in genomic DNA alkylations, with more MC-DNA adducts being formed when the enzyme was expressed closer to its DNA target. No change was observed in the hypoxic/aerobic differential toxicity as a function of enzyme localization. These findings indicate that drug efficacy is increased when the subcellular site of drug activation corresponds to its site of action. PMID- 15547248 TI - Negatively charged self-assembling DNA/poloxamine nanospheres for in vivo gene transfer. AB - Over the past decade, numerous nonviral cationic vectors have been synthesized. They share a high density of positive charges and efficiency for gene transfer in vitro. However, their positively charged surface causes instability in body fluids and cytotoxicity, thereby limiting their efficacy in vivo. Therefore, there is a need for developing alternative molecular structures. We have examined tetrabranched amphiphilic block copolymers consisting of four polyethyleneoxide/polypropyleneoxide blocks centered on an ethylenediamine moiety. Cryo-electron microscopy, ethidium bromide fluorescence and light and X ray scattering experiments performed on vector-DNA complexes showed that the dense core of the nanosphere consisted of condensed DNA interacting with poloxamine molecules through electrostatic, hydrogen bonding and hydrophobic interactions, with DNA molecules also being exposed at the surface. The supramolecular organization of block copolymer/DNA nanospheres induced the formation of negatively charged particles. These particles were stable in a solution that had a physiological ionic composition and were resistant to decomplexation by heparin. The new nanostructured material, the structure of which clearly contrasted with that of lipoplexes and polyplexes, efficiently transferred reporter and therapeutic genes in skeletal and heart muscle in vivo. Negatively charged supramolecular assemblies hold promise as therapeutic gene carriers for skeletal and heart muscle-related diseases and expression of therapeutic proteins for local or systemic uses. PMID- 15547249 TI - Identification of a novel human nuclear-encoded mitochondrial poly(A) polymerase. AB - We report here on the identification of a novel human nuclear-encoded mitochondrial poly(A) polymerase. Immunocytochemical experiments confirm that the enzyme indeed localizes to mitochondrial compartment. Inhibition of expression of the enzyme by RNA interference results in significant shortening of the poly(A) tails of the mitochondrial ND3, COX III and ATP 6/8 transcripts, suggesting that the investigated protein represents a bona fide mitochondrial poly(A) polymerase. This is in agreement with our sequencing data which show that poly(A) tails of several mitochondrial messengers are composed almost exclusively of adenosine residues. Moreover, the data presented here indicate that all analyzed mitochondrial transcripts with profoundly shortened poly(A) tails are relatively stable, which in turn argues against the direct role of long poly(A) extensions in the stabilization of human mitochondrial messengers. PMID- 15547250 TI - Transient overexpression of mitochondrial transcription factor A (TFAM) is sufficient to stimulate mitochondrial DNA transcription, but not sufficient to increase mtDNA copy number in cultured cells. AB - Mitochondrial transcription factor A (TFAM) stimulates transcription from mitochondrial DNA (mtDNA) promoters in vitro and in organello. To investigate whether changes of TFAM levels also modulate transcription and replication in situ, the protein was transiently overexpressed in cultured cells. Mitochondrial mRNAs were significantly elevated at early time points, when no expansion of the TFAM pool was yet observed, but were decreased when TFAM levels had doubled, resemb-ling in vitro results. HEK cells contain about 35 molecules of TFAM per mtDNA. High levels of TFAM were not associated with increases of full-length mtDNA, but nucleic acid species sensitive to RNAse H increased. Stimulation of transcription was more evident when TFAM was transiently overexpressed in cells pre-treated with ethidium bromide (EBr) having lowered mtDNA, TFAM and mitochondrial transcript levels. EBr rapidly inhibited mtDNA transcription, while decay of mtDNA was delayed and preferentially slowly migrating, relaxed mtDNA species were depleted. In conclusion, we show that transcription of mtDNA is submaximal in cultured cells and that a subtle increase of TFAM within the matrix is sufficient to stimulate mitochondrial transcription. Thus, this protein meets all criteria for being a key factor regulating mitochondrial transcription in vivo, but other factors are necessary for increasing mtDNA copy number, at least in cultured cells. PMID- 15547251 TI - A physiological connection between tmRNA and peptidyl-tRNA hydrolase functions in Escherichia coli. AB - The bacterial ssrA gene codes for a dual function RNA, tmRNA, which possesses tRNA-like and mRNA-like regions. The tmRNA appends an oligopeptide tag to the polypeptide on the P-site tRNA by a trans-translation process that rescues ribosomes stalled on the mRNAs and targets the aberrant protein for degradation. In cells, processing of the stalled ribosomes is also pioneered by drop-off of peptidyl-tRNAs. The ester bond linking the peptide to tRNA is hydrolyzed by peptidyl-tRNA hydrolase (Pth), an essential enzyme, which releases the tRNA and the aberrant peptide. As the trans-translation mechanism utilizes the peptidyl transferase activity of the stalled ribosomes to free the tRNA (as opposed to peptidyl-tRNA drop-off), the need for Pth to recycle such tRNAs is bypassed. Thus, we hypothesized that tmRNA may rescue a defect in Pth. Here, we show that overexpression of tmRNA rescues the temperature-sensitive phenotype of Escherichia coli (pth(ts)). Conversely, a null mutation in ssrA enhances the temperature-sensitive phenotype of the pth(ts) strain. Consistent with our hypothesis, overexpression of tmRNA results in decreased accumulation of peptidyl tRNA in E.coli. Furthermore, overproduction of tmRNA in E.coli strains deficient in ribosome recycling factor and/or lacking the release factor 3 enhances the rescue of pth(ts) strains. We discuss the physiological relevance of these observations to highlight a major role of tmRNA in decreasing cellular peptidyl tRNA load. PMID- 15547252 TI - Comparative analysis of the Borrelia garinii genome. AB - Three members of the genus Borrelia (B.burgdorferi, B.garinii, B.afzelii) cause tick-borne borreliosis. Depending on the Borrelia species involved, the borreliosis differs in its clinical symptoms. Comparative genomics opens up a way to elucidate the underlying differences in Borrelia species. We analysed a low redundancy whole-genome shotgun (WGS) assembly of a B.garinii strain isolated from a patient with neuroborreliosis in comparison to the B.burgdorferi genome. This analysis reveals that most of the chromosome is conserved (92.7% identity on DNA as well as on amino acid level) in the two species, and no chromosomal rearrangement or larger insertions/deletions could be observed. Furthermore, two collinear plasmids (lp54 and cp26) seem to belong to the basic genome inventory of Borrelia species. These three collinear parts of the Borrelia genome encode 861 genes, which are orthologous in the two species examined. The majority of the genetic information of the other plasmids of B.burgdorferii is also present in B.garinii although orthology is not easy to define due to a high redundancy of the plasmid fraction. Yet, we did not find counterparts of the B.burgdorferi plasmids lp36 and lp38 or their respective gene repertoire in the B.garinii genome. Thus, phenotypic differences between the two species could be attributable to the presence or absence of these two plasmids as well as to the potentially positively selected genes. PMID- 15547253 TI - Genetic characterization of the bile salt response in Lactobacillus plantarum and analysis of responsive promoters in vitro and in situ in the gastrointestinal tract. AB - In this paper we describe the growth, morphological, and genetic responses of Lactobacillus plantarum WCFS1 to bile. Growth experiments revealed that a stepwise increase in the porcine bile concentration led to a gradual decrease in the maximal growth rate. Moreover, the final density reached by an L. plantarum culture growing in MRS containing 0.1% bile was approximately threefold lower than that in MRS lacking bile. The morphology of the cells grown in MRS containing 0.1% bile was investigated by scanning electron microscopy, which revealed that cells clumped together and had rough surfaces and that some of the cells had a shrunken and empty appearance, which clearly contrasted with the characteristic rod-shaped, smooth-surface morphology of L. plantarum cells grown in MRS without bile. An alr complementation-based genome-wide promoter screening analysis was performed with L. plantarum, which led to identification of 31 genes whose expression was potentially induced by 0.1% porcine bile. Remarkably, 11 membrane- and cell wall-associated functions appeared to be induced by bile, as were five functions involved in redox reactions and five regulatory factors. Moreover, the lp_0237 and lp_0775 genes, identified here as genes that are inducible by bile in vitro, were previously identified in our laboratory as important for L. plantarum in vivo during passage in the mouse gastrointestinal tract (P. A. Bron, C. Grangette, A. Mercenier, W. M. de Vos, and M. Kleerebezem, J. Bacteriol. 186:5721-5729, 2004). A quantitative reverse transcription-PCR approach focusing on these two genes confirmed that the expression level of lp_0237 and lp_0775 was significantly higher in cells grown in the presence of bile and cells isolated from the mouse duodenum than in cells grown on laboratory medium without bile. PMID- 15547254 TI - Genetic dissection of the light-inducible carQRS promoter region of Myxococcus xanthus. AB - In Myxococcus xanthus photoprotective carotenoids are produced in response to illumination due to regulated expression of carotenoid biosynthesis genes at two loci. Induction of the carotenogenesis regulon is dependent on expression of the carQRS operon. The first gene product of the operon, CarQ, is a sigma factor belonging to the ECF family and is responsible for light-dependent initiation of transcription at the carQRS promoter. We defined the minimal carQRS promoter as a 145-bp fragment of DNA upstream of the carQRS transcriptional start site, which includes the promoter for a divergent gene, gufA. In order to elucidate regions with the promoter required for activity, point mutations were introduced into the carQRS promoter between positions -151 and 6. While most sequence changes abolished carQRS promoter activity, two changes enhanced promoter activity and two changes caused the mutant promoter to become constitutive and independent of CarQ. The promoter-null point mutations and 6-bp deletion mutations implied that the carQRS promoter requires a functional gufA promoter for transcriptional activity and vice versa. By mapping the extent of the promoter region, identifying sequences important for promoter activity, and highlighting potential topological effects, we provide a foundation for further analysis of the carQRS promoter. PMID- 15547255 TI - Transcriptional activation of sclA by Mga requires a distal binding site in Streptococcus pyogenes. AB - Streptococcus pyogenes (the group A streptococcus [GAS]) is a medically significant pathogen of humans, causing a range of diseases from pharyngitis to necrotizing fasciitis. Several important GAS virulence genes are under the control of a pleiotropic regulator called Mga, or the multiple gene regulator of GAS, including the gene encoding the streptococcal collagen-like protein, or sclA. Analysis of the genome sequence upstream of sclA revealed two potential Mga binding sites with homology to the published Mga-binding element, which were called PsclA-I (distal) and PsclA-II (proximal) based on their location relative to a predicted start of transcription. Primer extension was used to confirm that the Mga-dependent transcriptional start site for sclA was located adjacent to the proximal PsclA-II binding site. By using overlapping PsclA promoter probes and purified Mga-His fusion protein, it was shown by electrophoretic mobility shift assays that, unlike other Mga-regulated promoters, Mga binds only to a distal DNA binding site (PsclA-I). Binding of Mga to PsclA-I could be competed with cold probes corresponding to known Mga-regulated promoters (Pemm, PscpA, and Pmga) but not with a nonspecific probe or the proximal PsclA-II fragment. With the use of a plasmid-based green fluorescent protein transcriptional reporter system, the full length PsclA was not sufficient to reproduce normal Mga-regulated activation. However, studies using a single-copy gusA transcriptional reporter system integrated at the native sclA chromosomal locus clearly demonstrated that the distal PsclA-I binding site is required for Mga regulation. Therefore, PsclA represents a new class of Mga-regulated promoters that requires a single distal binding site for activation. PMID- 15547256 TI - Binding of the C-terminal domain of the alpha subunit of RNA polymerase to the phage mu middle promoter. AB - The C-terminal domain of the alpha subunit (alpha CTD) of Escherichia coli RNA polymerase is often involved in transcriptional regulation. The alpha CTD typically stimulates transcription via interactions with promoter UP element DNA and transcriptional activators. DNase I footprinting and gel mobility shift assays were used to look for potential interaction of the alpha CTD with the phage Mu middle promoter P(m) and its activator protein Mor. Binding of RNA polymerase to P(m) in the presence of Mor resulted in production of a DNase I footprint downstream of Mor due to open complex formation and generation of a second footprint just upstream of the Mor binding site. Generation of the upstream footprint did not require open complex formation and also occurred in reactions in which the alpha CTD or His-alpha proteins were substituted for RNA polymerase. In gel mobility shift assays, the formation of a supershifted ternary complex demonstrated that Mor and His-alpha bind synergistically to P(m) DNA. Gel shift assays with short DNA fragments demonstrated that only the Mor binding site and a single upstream alpha CTD binding site were required for ternary complex formation. These results suggest that the alpha CTD plays a role in P(m) transcription by binding to P(m) DNA just upstream from Mor and making protein protein interactions with Mor that stabilize the binding of both proteins. PMID- 15547257 TI - Teichoic acid is an essential polymer in Bacillus subtilis that is functionally distinct from teichuronic acid. AB - Wall teichoic acids are anionic, phosphate-rich polymers linked to the peptidoglycan of gram-positive bacteria. In Bacillus subtilis, the predominant wall teichoic acid types are poly(glycerol phosphate) in strain 168 and poly(ribitol phosphate) in strain W23, and they are synthesized by the tag and tar gene products, respectively. Growing evidence suggests that wall teichoic acids are essential in B. subtilis; however, it is widely believed that teichoic acids are dispensable under phosphate-limiting conditions. In the work reported here, we carefully studied the dispensability of teichoic acid under phosphate limiting conditions by constructing three new mutants. These strains, having precise deletions in tagB, tagF, and tarD, were dependent on xylose-inducible complementation from a distal locus (amyE) for growth. The tarD deletion interrupted poly(ribitol phosphate) synthesis in B. subtilis and represents a unique deletion of a tar gene. When teichoic acid biosynthetic proteins were depleted, the mutants showed a coccoid morphology and cell wall thickening. The new wall teichoic acid biogenesis mutants generated in this work and a previously reported tagD mutant were not viable under phosphate-limiting conditions in the absence of complementation. Cell wall analysis of B. subtilis grown under phosphate-limited conditions showed that teichoic acid contributed approximately one-third of the wall anionic content. These data suggest that wall teichoic acid has an essential function in B. subtilis that cannot be replaced by teichuronic acid. PMID- 15547258 TI - KatG is the primary detoxifier of hydrogen peroxide produced by aerobic metabolism in Bradyrhizobium japonicum. AB - Bacteria are exposed to reactive oxygen species from the environment and from those generated by aerobic metabolism. Catalases are heme proteins that detoxify H(2)O(2), and many bacteria contain more than one catalase enzyme. Also, the nonheme peroxidase alkyl hydroperoxide reductase (Ahp) is the major scavenger of endogenous H(2)O(2) in Escherichia coli. Here, we show that aerobically grown Bradyrhizobium japonicum cells express a single catalase activity. Four genes encoding putative catalases in the B. japonicum genome were identified, including a katG homolog encoding a catalase-peroxidase. Deletion of the katG gene resulted in loss of catalase activity in cell extracts and of exogenous H(2)O(2) consumption by whole cells. The katG strain had a severe aerobic growth phenotype but showed improved growth in the absence of O(2). By contrast, a B. japonicum ahpCD mutant grew well aerobically and consumed H(2)O(2) at wild-type rates. A heme-deficient hemA mutant expressed about one-third of the KatG activity as the wild type but grew well aerobically and scavenged low concentrations of exogenous H(2)O(2). However, cells of the hemA strain were deficient in consumption of high concentrations of H(2)O(2) and were very sensitive to killing by short exposure to H(2)O(2). In addition, KatG activity did not decrease as a result of mutation of the gene encoding the transcriptional activator OxyR. We conclude that aerobic metabolism produces toxic levels of H(2)O(2) in B. japonicum, which is detoxified primarily by KatG. Furthermore, the katG level sufficient for detoxification does not require OxyR. PMID- 15547259 TI - Purification and initial characterization of the Salmonella enterica PduO ATP:Cob(I)alamin adenosyltransferase. AB - The PduO enzyme of Salmonella enterica is an ATP:cob(I)alamin adenosyltransferase that catalyzes the final step in the conversion of vitamin B(12) to coenzyme B(12). The primary physiological role of this enzyme is to support coenzyme B(12) dependent 1,2-propanediol degradation, and bioinformatic analysis has indicated that it has two domains. Here the PduO adenosyltransferase was produced in Escherichia coli, solubilized from inclusion bodies, purified to apparent homogeneity, and partially characterized biochemically. The K(m) values of PduO for ATP and cob(I)alamin were 19.8 and 4.5 microM, respectively, and the enzyme V(max) was 243 nmol min(-1) mg of protein(-1). Further investigations showed that PduO was active with ATP and partially active with deoxy-ATP, but lacked measurable activity with other nucleotides. (31)P nuclear magnetic resonance established that triphosphate was a product of the PduO reaction, and kinetic studies indicated a ternary complex mechanism. A series of truncated versions of the PduO protein were produced in Escherichia coli, partially purified, and used to show that adenosyltransferase activity is associated with the N-terminal domain. The N-terminal domain was purified to near homogeneity and shown to have biochemical properties and kinetic constants similar to those of the full-length enzyme. This indicated that the C-terminal domain was not directly involved in catalysis or substrate binding and may have another role. PMID- 15547260 TI - Rubrerythrin from the hyperthermophilic archaeon Pyrococcus furiosus is a rubredoxin-dependent, iron-containing peroxidase. AB - Rubrerythrin was purified by multistep chromatography under anaerobic, reducing conditions from the hyperthermophilic archaeon Pyrococcus furiosus. It is a homodimer with a molecular mass of 39.2 kDa and contains 2.9 +/- 0.2 iron atoms per subunit. The purified protein had peroxidase activity at 85 degrees C using hydrogen peroxide with reduced P. furiosus rubredoxin as the electron donor. The specific activity was 36 micromol of rubredoxin oxidized/min/mg with apparent K(m) values of 35 and 70 microM for hydrogen peroxide and rubredoxin, respectively. When rubrerythrin was combined with rubredoxin and P. furiosus NADH:rubredoxin oxidoreductase, the complete system used NADH as the electron donor to reduce hydrogen peroxide with a specific activity of 7.0 micromol of H(2)O(2) reduced/min/mg of rubrerythrin at 85 degrees C. Strangely, as-purified (reduced) rubrerythrin precipitated when oxidized by either hydrogen peroxide, air, or ferricyanide. The gene (PF1283) encoding rubrerythrin was expressed in Escherichia coli grown in medium with various metal contents. The purified recombinant proteins each contained approximately three metal atoms/subunit, ranging from 0.4 Fe plus 2.2 Zn to 1.9 Fe plus 1.2 Zn, where the metal content of the protein depended on the metal content of the E. coli growth medium. The peroxidase activities of the recombinant forms were proportional to the iron content. P. furiosus rubrerythrin is the first to be characterized from a hyperthermophile or from an archaeon, and the results are the first demonstration that this protein functions in an NADH-dependent, hydrogen peroxide:rubredoxin oxidoreductase system. Rubrerythrin is proposed to play a role in the recently defined anaerobic detoxification pathway for reactive oxygen species. PMID- 15547261 TI - Ralstonia solanacearum iron scavenging by the siderophore staphyloferrin B is controlled by PhcA, the global virulence regulator. AB - PhcA is a transcriptional regulator that activates expression of multiple virulence genes in the plant pathogen Ralstonia solanacearum. Relative to their wild-type parents, phcA mutants overproduced iron-scavenging activity detected with chrome azurol S siderophore detection medium. Transposon mutagenesis of strain AW1-PC (phcA1) generated strain GB6, which was siderophore negative but retained weak iron-scavenging activity. The ssd gene inactivated in GB6 encodes a protein similar to group IV amino acid decarboxylases, and its transcription was repressed by iron(III) and PhcA. ssd is the terminal gene in a putative operon that also appears to encode three siderophore synthetase subunits, a integral membrane exporter, and three genes with no obvious role in siderophore production. A homologous operon was found in the genomes of Ralstonia metallidurans and Staphylococcus aureus, both of which produce the polycarboxylate siderophore staphyloferrin B. Comparison of the siderophores present in culture supernatants of R. solanacearum, R. metallidurans, and Bacillus megaterium using chemical tests, a siderophore utilization bioassay, thin-layer chromatography, and mass spectroscopy indicated that R. solanacearum produces staphyloferrin B rather than schizokinen as was reported previously. Inactivation of ssd in a wild-type AW1 background resulted in a mutant almost incapable of scavenging iron but normally virulent on tomato plants. AW1 did not produce siderophore activity when cultured in tomato xylem sap, suggesting that the main location in tomato for R. solanacearum during pathogenesis is iron replete. PMID- 15547262 TI - The recombination genes addAB are not restricted to gram-positive bacteria: genetic analysis of the recombination initiation enzymes RecF and AddAB in Rhizobium etli. AB - Single-strand gaps (SSGs) and double-strand breaks (DSBs) are the major initiation sites for recombination. In bacteria, the SSGs are repaired by RecFOR, while the DSBs are processed by RecBCD in gram-negative bacteria and AddAB in gram-positive bacteria. Unexpectedly, instead of recBCD genes, the addAB genes were found in members of the alpha-proteobacteria group (gram negative). Taking Rhizobium etli as a model, the role of recF and addAB genes in homologous recombination and repair of damaged DNA was evaluated. Inactivation of either recF or addA provoked strong sensitivity to UV radiation and mitomycin C, while an additive effect was observed in the recF-addA mutant. The DSBs generated by nalidixic acid caused low viability only in the addA mutant. The recombination frequency of large and small plasmids was reduced in the recF mutant (24- and 36 fold, respectively), whereas a slight decrease (threefold) in the addA mutant was observed. Moreover, an additive effect (47- and 90-fold, respectively) was observed in the double mutant, but it was not as dramatic as that in a recA mutant. Interestingly, the frequency of deletion and Campbell-type recombination was slightly affected in either single or double mutants. These results suggest that another pathway exists that allows plasmid and Campbell-type recombination in the absence of recF and addA genes. PMID- 15547263 TI - Positive and negative transcriptional regulators of glutathione-dependent formaldehyde metabolism. AB - A glutathione (GSH)-dependent pathway is used for formaldehyde metabolism by a wide variety of prokaryotes and eukaryotes. In this pathway, S hydroxymethylglutathione, produced by the reaction of formaldehyde with the thiolate moiety of glutathione, is the substrate for a GSH-dependent formaldehyde dehydrogenase (GSH-FDH). While expression of GSH-FDH often increases in the presence of metabolic or exogenous sources of formaldehyde, little is known about the factors that regulate this response. Here, we identify two signal transduction pathways that regulate expression of adhI, the gene encoding GSH FDH, in Rhodobacter sphaeroides. The loss of the histidine kinase response regulator pair RfdRS or the histidine kinase RfdS increases adhI transcription in the absence of metabolic sources of formaldehyde. Cells lacking RfdRS further increase adhI expression in the presence of metabolic sources of formaldehyde (methanol), suggesting that this negative regulator of GSH-FDH expression does not respond to this compound. In contrast, mutants lacking the histidine kinase response regulator pair AfdRS or the histidine kinase AfdS cannot induce adhI expression in the presence of either formaldehyde or metabolic sources of this compound. AfdR stimulates activity of the adhI promoter in vitro, indicating that this protein is a direct activator of GSH-FDH expression. Activation by AfdR is detectable only after incubation of the protein with acetyl phosphate, suggesting that phosphorylation is necessary for transcription activation. Activation of adhI transcription by acetyl-phosphate-treated AfdR in vitro is inhibited by a truncated RfdR protein, suggesting that this protein is a direct repressor of GSH FDH expression. Together, the data indicate that AfdRS and RfdRS positively and negatively regulate adhI transcription in response to different signals. PMID- 15547264 TI - Global transposon mutagenesis and essential gene analysis of Helicobacter pylori. AB - We have constructed a genome-saturating mutant library of the human gastric pathogen Helicobacter pylori. Microarray tracking of transposon mutants (MATT) allowed us to map the position of 5,363 transposon mutants in our library. While we generally found insertions well distributed throughout the genome, 344 genes had no detectable transposon insertions, and this list is predicted to be highly enriched for essential genes. Comparison to the essential gene set of other bacteria revealed a surprisingly limited overlap with all organisms tested (11%), while 55% were essential in some organisms but not others. We independently verified the essentiality of several gene products, including an HtrA family serine protease, a hypothetical protein with putative phospholipase D activity, and a riboflavin specific deaminase. A limited screen for motility mutants allowed us to estimate that 4.5% of the genome is dedicated to this virulence associated phenotype. PMID- 15547265 TI - In vivo activity of enzymatic and regulatory components of the phosphoenolpyruvate:sugar phosphotransferase system in Mycoplasma pneumoniae. AB - Mycoplasma pneumoniae is a pathogenic bacterium that is highly adapted to life on mucosal surfaces. This adaptation is reflected by the very compact genome and the small number of regulatory proteins. However, M. pneumoniae possesses the HPr kinase/phosphorylase (HPrK/P), the key regulator of carbon metabolism in the Firmicutes. In contrast to the enzymes of other bacteria, the HPrK/P of M. pneumoniae is already active at very low ATP concentrations, suggesting a different mode of regulation. In this work, we studied the ability of M. pneumoniae to utilize different carbohydrates and their effects on the activity of the different phosphotransferase system (PTS) components. Glucose served as the best carbon source, with a generation time of about 30 h. Fructose and glycerol were also used but at lower rates and with lower yields. In contrast, M. pneumoniae is unable to use mannitol even though the bacterium is apparently equipped with all the genes required for mannitol catabolism. This observation is probably a reflection of the continuing and ongoing reduction of the M. pneumoniae genome. The general enzymatic and regulatory components of the PTS, i.e., enzyme I, HPr, and HPrK/P, were present under all growth conditions tested in this study. However, HPrK/P activity is strongly increased if the medium contains glycerol. Thus, the control of HPrK/P in vivo differs strongly between M. pneumoniae and the other Firmicutes. This difference may relate to the specific conditions on lipid-rich cell surfaces. PMID- 15547266 TI - Physiological and gene expression analysis of inhibition of Desulfovibrio vulgaris hildenborough by nitrite. AB - A Desulfovibrio vulgaris Hildenborough mutant lacking the nrfA gene for the catalytic subunit of periplasmic cytochrome c nitrite reductase (NrfHA) was constructed. In mid-log phase, growth of the wild type in medium containing lactate and sulfate was inhibited by 10 mM nitrite, whereas 0.6 mM nitrite inhibited the nrfA mutant. Lower concentrations (0.04 mM) inhibited the growth of both mutant and wild-type cells on plates. Macroarray hybridization indicated that nitrite upregulates the nrfHA genes and downregulates genes for sulfate reduction enzymes catalyzing steps preceding the reduction of sulfite to sulfide by dissimilatory sulfite reductase (DsrAB), for two membrane-bound electron transport complexes (qmoABC and dsrMKJOP) and for ATP synthase (atp). DsrAB is known to bind and slowly reduce nitrite. The data support a model in which nitrite inhibits DsrAB (apparent dissociation constant K(m) for nitrite = 0.03 mM), and in which NrfHA (K(m) for nitrite = 1.4 mM) limits nitrite entry by reducing it to ammonia when nitrite concentrations are at millimolar levels. The gene expression data and consideration of relative gene locations suggest that QmoABC and DsrMKJOP donate electrons to adenosine phosphosulfate reductase and DsrAB, respectively. Downregulation of atp genes, as well as the recorded cell death following addition of inhibitory nitrite concentrations, suggests that the proton gradient collapses when electrons are diverted from cytoplasmic sulfate to periplasmic nitrite reduction. PMID- 15547267 TI - Systematic inactivation and phenotypic characterization of two-component signal transduction systems of Enterococcus faecalis V583. AB - The ability of enterococci to adapt and respond to different environmental stimuli, including the host environment, led us to investigate the role of two component signal transduction in the regulation of Enterococcus faecalis physiology. Using a bioinformatic approach, we previously identified 17 two component systems (TCS), consisting of a sensory histidine kinase and the cognate response regulator, as well as an additional orphan response regulator (L. E. Hancock and M. Perego, J. Bacteriol. 184:5819-5825, 2002). In an effort to identify the potential function of each TCS in the biology of E. faecalis clinical isolate strain V583, we constructed insertion mutations in each of the response regulators. We were able to inactivate 17 of 18 response regulators, the exception being an ortholog of YycF, previously shown to be essential for viability in a variety of gram-positive microorganisms. The biological effects of the remaining mutations were assessed by using a number of assays, including antibiotic resistance, biofilm formation, and environmental stress. We identified TCS related to antibiotic resistance and environmental stress and found one system which controls the initiation of biofilm development by E. faecalis. PMID- 15547268 TI - Population structure and evolution of the Bacillus cereus group. AB - Representative strains of the Bacillus cereus group of bacteria, including Bacillus anthracis (11 isolates), B. cereus (38 isolates), Bacillus mycoides (1 isolate), Bacillus thuringiensis (53 isolates from 17 serovars), and Bacillus weihenstephanensis (2 isolates) were assigned to 59 sequence types (STs) derived from the nucleotide sequences of seven alleles, glpF, gmk, ilvD, pta, pur, pycA, and tpi. Comparisons of the maximum likelihood (ML) tree of the concatenated sequences with individual gene trees showed more congruence than expected by chance, indicating a generally clonal structure to the population. The STs followed two major lines of descent. Clade 1 comprised B. anthracis strains, numerous B. cereus strains, and rare B. thuringiensis strains, while clade 2 included the majority of the B. thuringiensis strains together with some B. cereus strains. Other species were allocated to a third, heterogeneous clade. The ML trees and split decomposition analysis were used to assign STs to eight lineages within clades 1 and 2. These lineages were defined by bootstrap analysis and by a preponderance of fixed differences over shared polymorphisms among the STs. Lineages were named with reference to existing designations: Anthracis, Cereus I, Cereus II, Cereus III, Kurstaki, Sotto, Thuringiensis, and Tolworthi. Strains from some B. thuringiensis serovars were wholly or largely assigned to a single ST, for example, serovar aizawai isolates were assigned to ST-15, serovar kenyae isolates were assigned to ST-13, and serovar tolworthi isolates were assigned to ST-23, while other serovars, such as serovar canadensis, were genetically heterogeneous. We suggest a revision of the nomenclature in which the lineage and clone are recognized through name and ST designations in accordance with the clonal structure of the population. PMID- 15547269 TI - Negative transcriptional regulation of the ilv-leu operon for biosynthesis of branched-chain amino acids through the Bacillus subtilis global regulator TnrA. AB - The Bacillus subtilis ilv-leu operon is involved in the synthesis of branched chain amino acids (valine, isoleucine, and leucine). The two- to threefold repression of expression of the ilv-leu operon during logarithmic-phase growth under nitrogen-limited conditions, which was originally detected by a DNA microarray analysis to compare the transcriptomes from the wild-type and tnrA mutant strains, was confirmed by lacZ fusion and Northern experiments. A genome wide TnrA box search revealed a candidate box approximately 200 bp upstream of the transcription initiation base of the ilv-leu operon, the TnrA binding to which was verified by gel retardation and DNase I footprinting analyses. Deletion and base substitution of the TnrA box sequence affected the ilv-leu promoter activity in vivo, implying that TnrA bound to the box might be able to inhibit the promoter activity, possibly through DNA bending. The negative control of the expression of the ilv-leu operon by TnrA, which is considered to represent rather fine-tuning (two- to threefold), is a novel regulatory link between nitrogen and amino acid metabolism. PMID- 15547270 TI - A DNA region recognized by the nitric oxide-responsive transcriptional activator NorR is conserved in beta- and gamma-proteobacteria. AB - The sigma(54)-dependent regulator NorR activates transcription of target genes in response to nitric oxide (NO) or NO-generating agents. In Ralstonia eutropha H16, NorR activates transcription of the dicistronic norAB operon that encodes NorA, a protein of unknown function, and NorB, a nitric oxide reductase. A constitutively activating NorR derivative (NorR'), in which the N-terminal signaling domain was replaced by MalE, specifically bound to the norAB upstream region as revealed by gel retardation analysis. Within a 73-bp DNA segment protected by MalE-NorR' in a DNase I footprint assay, three conserved inverted repeats, GGT-(N(7))-ACC (where N is any base), that we consider to be NorR-binding boxes were identified. Mutations altering the spacing or the base sequence of these repeats resulted in an 80 to 90% decrease of transcriptional activation by wild-type NorR. Genome database analyses demonstrate that the GT-(N(7))-AC core of the inverted repeat is found in several proteobacteria upstream of gene loci encoding proteins of nitric oxide metabolism, including nitric oxide reductase (NorB), flavorubredoxin (NorV), NO dioxygenase (Hmp), and hybrid cluster protein (Hcp). PMID- 15547271 TI - Sequence tolerance of the phage lambda PRM promoter: implications for evolution of gene regulatory circuitry. AB - Much of the gene regulatory circuitry of phage lambda centers on a complex region called the O(R) region. This approximately 100-bp region is densely packed with regulatory sites, including two promoters and three repressor-binding sites. The dense packing of this region is likely to impose severe constraints on its ability to change during evolution, raising the question of how the specific arrangement of sites and their exact sequences could evolve to their present form. Here we ask whether the sequence of a cis-acting site can be widely varied while retaining its function; if it can, evolution could proceed by a larger number of paths. To help address this question, we developed a lambda cloning vector that allowed us to clone fragments spanning the O(R) region. By using this vector, we carried out intensive mutagenesis of the P(RM) promoter, which drives expression of CI repressor and is activated by CI itself. We made a pool of fragments in which 8 of the 12 positions in the -35 and -10 regions were randomized and cloned this pool into the vector, making a pool of P(RM) variant phage. About 10% of the P(RM) variants were able to lysogenize, suggesting that the lambda regulatory circuitry is compatible with a wide range of P(RM) sequences. Analysis of several of these phages indicated a range of behaviors in prophage induction. Several isolates had induction properties similar to those of the wild type, and their promoters resembled the wild type in their responses to CI. We term this property of different sequences allowing roughly equivalent function "sequence tolerance " and discuss its role in the evolution of gene regulatory circuitry. PMID- 15547272 TI - Two outer membrane proteins are required for maximal type I secretion of the Caulobacter crescentus S-layer protein. AB - Transport of RsaA, the crystalline S-layer subunit protein of Caulobacter crescentus, is mediated by a type I secretion mechanism. Two proteins have been identified that play the role of the outer membrane protein (OMP) component in the RsaA secretion machinery. The genes rsaF(a) and rsaF(b) were identified by similarity to the Escherichia coli hemolysin secretion OMP TolC by using the C. crescentus genome sequence. The rsaF(a) gene is located several kilobases downstream of the other transporter genes, while rsaF(b) is completely unlinked. An rsaF(a) knockout had approximately 56% secretion compared to wild-type levels, while the rsaF(b) knockout reduced secretion levels to approximately 79%. When expression of both proteins was eliminated, there was no RsaA secretion, but a residual level of approximately 9% remained inside the cell, suggesting posttranslational autoregulation. Complementation with either of the individual rsaF genes by use of a multicopy vector, which resulted in 8- to 10-fold overexpression of the proteins, did not restore RsaA secretion to wild-type levels, indicating that both rsaF genes were required for full-level secretion. However, overexpression of rsaF(a) (with normal rsaF(b) levels) in concert with overexpression of rsaA resulted in a 28% increase in RsaA secretion, indicating a potential for significantly increasing expression levels of an already highly expressing type I secretion system. This is the only known example of type I secretion requiring two OMPs to assemble a fully functional system. PMID- 15547273 TI - Essentiality of the early transcript in the replication origin of the lactococcal prolate phage c2. AB - The genome of the prolate-headed lytic lactococcal bacteriophage c2 is organized into two divergently oriented blocks consisting of the early genes and the late genes. These blocks are separated by the noncoding origin of DNA replication. We examined the functional role of transcription of the origin in a plasmid model system. Deletion of the early promoter P(E)1 abolished origin function. Introduction of mutations into P(E)1 which did not eliminate promoter activity or replacement of P(E)1 with an unrelated but functional promoter did not abolish replication. The A-T-rich region upstream of P(E)1, which is conserved in prolate phages, was not required for plasmid replication. Replacement of the P(E)1 transcript template sequence with an unrelated sequence with a similar G+C content abolished replication, showing that the sequence encoding the transcript is essential for origin function. Truncated transcript and internal deletion constructs did not support replication except when the deletion was at the very 3' end of the DNA sequence coding for the transcript. The P(E)1 transcript could be detected for all replication-proficient constructs. Recloning in a plasmid vector allowed detection of P(E)1 transcripts from some fragments that did not support replication, indicating that stability of the transcript alone was not sufficient for replication. The data suggest that production of a transcript of a specific length and with a specific sequence or structure is essential for the function of the phage c2 origin in this model system. PMID- 15547274 TI - Kinetic analysis of the oxidative conversion of the [4Fe-4S]2+ cluster of FNR to a [2Fe-2S]2+ Cluster. AB - The ability of FNR to sense and respond to cellular O(2) levels depends on its [4Fe-4S](2+) cluster. In the presence of O(2), the [4Fe-4S](2+) cluster is converted to a [2Fe-2S](2+) cluster, which inactivates FNR as a transcriptional regulator. In this study, we demonstrate that approximately 2 Fe(2+) ions are released from the reaction of O(2) with the [4Fe-4S](2+) cluster. Fe(2+) release was then used as an assay of reaction progress to investigate the rate of [4Fe 4S](2+) to [2Fe-2S](2+) cluster conversion in vitro. We also found that there was no detectable difference in the rate of O(2)-induced cluster conversion for FNR free in solution compared to its DNA-bound form. In addition, the rate of FNR inactivation was monitored in vivo by measuring the rate at which transcriptional regulation by FNR is lost upon the exposure of cells to O(2); a comparison of the in vitro and in vivo rates of conversion suggests that O(2)-induced cluster conversion is sufficient to explain FNR inactivation in cells. FNR protein levels were also compared for cells grown under aerobic and anaerobic conditions. PMID- 15547275 TI - Effector-mediated interaction of CbbRI and CbbRII regulators with target sequences in Rhodobacter capsulatus. AB - In Rhodobacter capsulatus, genes encoding enzymes of the Calvin-Benson-Bassham reductive pentose phosphate pathway are located in the cbb(I) and cbb(II) operons. Each operon contains a divergently transcribed LysR-type transcriptional activator (CbbR(I) and CbbR(II)) that regulates the expression of its cognate cbb promoter in response to an as yet unidentified effector molecule(s). Both CbbR(I) and CbbR(II) were purified, and the ability of a variety of potential effector molecules to induce changes in their DNA binding properties at their target promoters was assessed. The responses of CbbR(I) and CbbR(II) to potential effectors were not identical. In gel mobility shift assays, the affinity of both CbbR(I) and CbbR(II) for their target promoters was enhanced in the presence of ribulose-1,5-bisphosphate (RuBP), phosphoenolpyruvate, 3-phosphoglycerate, 2 phosphoglycolate. ATP, 2-phosphoglycerate, and KH(2)PO(4) were found to enhance only CbbR(I) binding, while fructose-1,6-bisphosphate enhanced the binding of only CbbR(II). The DNase I footprint of CbbR(I) was reduced in the presence of RuBP, while reductions in the CbbR(II) DNase I footprint were induced by fructose 1,6-bisphosphate, 3-phosphoglycerate, and KH(2)PO(4). The current in vitro results plus recent in vivo studies suggest that CbbR-mediated regulation of cbb transcription is controlled by multiple metabolic signals in R. capsulatus. This control reflects not only intracellular levels of Calvin-Benson-Bassham cycle metabolic intermediates but also the fixed (organic) carbon status and energy charge of the cell. PMID- 15547276 TI - The chromosomally encoded cation diffusion facilitator proteins DmeF and FieF from Wautersia metallidurans CH34 are transporters of broad metal specificity. AB - Genomic sequencing of the beta-proteobacterium Wautersia (previously Ralstonia) metallidurans CH34 revealed the presence of three genes encoding proteins of the cation diffusion facilitator (CDF) family. One, CzcD, was previously found to be part of the high-level metal resistance system Czc that mediates the efflux of Co(II), Zn(II), and Cd(II) ions catalyzed by the CzcCBA cation-proton antiporter. The second CDF protein, FieF, is probably mainly a ferrous iron detoxifying protein but also mediated some resistance against other divalent metal cations such as Zn(II), Co(II), Cd(II), and Ni(II) in W. metallidurans or Escherichia coli. The third CDF protein, DmeF, showed the same substrate spectrum as FieF, but with different preferences. DmeF plays the central role in cobalt homeostasis in W. metallidurans, and a disruption of dmeF rendered the high-level metal cation resistance systems Czc and Cnr ineffective against Co(II). This is evidence for the periplasmic detoxification of substrates by RND transporters of the heavy metal efflux family subgroup. PMID- 15547277 TI - Phenylphosphate synthase: a new phosphotransferase catalyzing the first step in anaerobic phenol metabolism in Thauera aromatica. AB - The anaerobic metabolism of phenol in the beta-proteobacterium Thauera aromatica proceeds via para-carboxylation of phenol (biological Kolbe-Schmitt carboxylation). In the first step, phenol is converted to phenylphosphate which is then carboxylated to 4-hydroxybenzoate in the second step. Phenylphosphate formation is catalyzed by the novel enzyme phenylphosphate synthase, which was studied. Phenylphosphate synthase consists of three proteins whose genes are located adjacent to each other on the phenol operon and were overproduced in Escherichia coli. The promoter region and operon structure of the phenol gene cluster were investigated. Protein 1 (70 kDa) resembles the central part of classical phosphoenolpyruvate synthase which contains a conserved histidine residue. It catalyzes the exchange of free [(14)C]phenol and the phenol moiety of phenylphosphate but not the phosphorylation of phenol. Phosphorylation of phenol requires protein 1, MgATP, and another protein, protein 2 (40 kDa), which resembles the N-terminal part of phosphoenol pyruvate synthase. Proteins 1 and 2 catalyze the following reaction: phenol + MgATP + H(2)O-->phenylphosphate + MgAMP + orthophosphate. The phosphoryl group in phenylphosphate is derived from the beta-phosphate group of ATP. The free energy of ATP hydrolysis obviously favors the trapping of phenol (K(m), 0.04 mM), even at a low ambient substrate concentration. The reaction is stimulated severalfold by another protein, protein 3 (24 kDa), which contains two cystathionine-beta-synthase domains of unknown function but does not show significant overall similarity to known proteins. The molecular and catalytic features of phenylphosphate synthase resemble those of phosphoenolpyruvate synthase, albeit with interesting modifications. PMID- 15547278 TI - Novel roles for the AIDA adhesin from diarrheagenic Escherichia coli: cell aggregation and biofilm formation. AB - Diarrhea-causing Escherichia coli strains are responsible for numerous cases of gastrointestinal disease and constitute a serious health problem throughout the world. The ability to recognize and attach to host intestinal surfaces is an essential step in the pathogenesis of such strains. AIDA is a potent bacterial adhesin associated with some diarrheagenic E. coli strains. AIDA mediates bacterial attachment to a broad variety of human and other mammalian cells. It is a surface-displayed autotransporter protein and belongs to the selected group of bacterial glycoproteins; only the glycosylated form binds to mammalian cells. Here, we show that AIDA possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrate that intercellular AIDA AIDA interaction is responsible for bacterial autoaggregation. Interestingly, AIDA-expressing cells can interact with antigen 43 (Ag43)-expressing cells, which is indicative of an intercellular AIDA-Ag43 interaction. Additionally, AIDA expression dramatically enhances biofilm formation by E. coli on abiotic surfaces in flow chambers. PMID- 15547279 TI - Bacteriophage and phenotypic variation in Pseudomonas aeruginosa biofilm development. AB - A current question in biofilm research is whether biofilm-specific genetic processes can lead to differentiation in physiology and function among biofilm cells. In Pseudomonas aeruginosa, phenotypic variants which exhibit a small colony phenotype on agar media and a markedly accelerated pattern of biofilm development compared to that of the parental strain are often isolated from biofilms. We grew P. aeruginosa biofilms in glass flow cell reactors and observed that the emergence of small-colony variants (SCVs) in the effluent runoff from the biofilms correlated with the emergence of plaque-forming Pf1-like filamentous phage (designated Pf4) from the biofilm. Because several recent studies have shown that bacteriophage genes are among the most highly upregulated groups of genes during biofilm development, we investigated whether Pf4 plays a role in SCV formation during P. aeruginosa biofilm development. We carried out immunoelectron microscopy using anti-Pf4 antibodies and observed that SCV cells, but not parental-type cells, exhibited high densities of Pf4 filaments on the cell surface and that these filaments were often tightly interwoven into complex latticeworks surrounding the cells. Moreover, infection of P. aeruginosa planktonic cultures with Pf4 caused the emergence of SCVs within the culture. These SCVs exhibited enhanced attachment, accelerated biofilm development, and large regions of dead and lysed cells inside microcolonies in a manner identical to that of SCVs obtained from biofilms. We concluded that Pf4 can mediate phenotypic variation in P. aeruginosa biofilms. We also performed partial sequencing and analysis of the Pf4 replicative form and identified a number of open reading frames not previously recognized in the genome of P. aeruginosa, including a putative postsegregational killing operon. PMID- 15547280 TI - Crystal structure of the PdxY Protein from Escherichia coli. AB - The crystal structure of Escherichia coli PdxY, the protein product of the pdxY gene, has been determined to a 2.2-A resolution. PdxY is a member of the ribokinase superfamily of enzymes and has sequence homology with pyridoxal kinases that phosphorylate pyridoxal at the C-5' hydroxyl. The protein is a homodimer with an active site on each monomer composed of residues that come exclusively from each respective subunit. The active site is filled with a density that fits that of pyridoxal. In monomer A, the ligand appears to be covalently attached to Cys122 as a thiohemiacetal, while in monomer B it is not covalently attached but appears to be partially present as pyridoxal 5' phosphate. The presence of pyridoxal phosphate and pyridoxal as ligands was confirmed by the activation of aposerine hydroxymethyltransferase after release of the ligand by the denaturation of PdxY. The ligand, which appears to be covalently attached to Cys122, does not dissociate after denaturation of the protein. A detailed comparison (of functional properties, sequence homology, active site and ATP-binding-site residues, and active site flap types) of PdxY with other pyridoxal kinases as well as the ribokinase superfamily in general suggested that PdxY is a member of a new subclass of the ribokinase superfamily. The structure of PdxY also permitted an interpretation of work that was previously published about this enzyme. PMID- 15547281 TI - Structural similarity of YbeD protein from Escherichia coli to allosteric regulatory domains. AB - Lipoic acid is an essential prosthetic group in several metabolic pathways. The biosynthetic pathway of protein lipoylation in Escherichia coli involves gene products of the lip operon. YbeD is a conserved bacterial protein located in the dacA-lipB intergenic region. Here, we report the nuclear magnetic resonance structure of YbeD from E. coli. The structure includes a beta alpha beta beta alpha beta fold with two alpha-helices on one side of a four-strand antiparallel beta-sheet. The beta 2-beta 3 loop shows the highest sequence conservation and is likely functionally important. The beta-sheet surface contains a patch of conserved hydrophobic residues, suggesting a role in protein-protein interactions. YbeD shows striking structural homology to the regulatory domain from d-3-phosphoglycerate dehydrogenase, hinting at a role in the allosteric regulation of lipoic acid biosynthesis or the glycine cleavage system. PMID- 15547282 TI - Unmasking novel sporulation genes in Bacillus subtilis. AB - The Bacillus subtilis transcription factor sigma(E) directs the expression of a regulon of 262 genes, but null mutations in only a small fraction of these genes severely impair sporulation. We have previously reported that mutations in seven sigma(E)-controlled genes cause a mild (2- to 10-fold) defect in sporulation. In this study, we found that pairwise combinations of some of these seven mutations led to strong synthetic sporulation phenotypes, especially those involving the ytrHI operon and ybaN. Double mutants of ybaN and ytrH and of ybaN and ytrI had >10,000-fold lower sporulation efficiencies than the wild type. Thin-section electron microscopy revealed a block in cortex formation for the ybaN ytrH double mutant and coat defects for the ybaN single and ybaN ytrI double mutants. Sporulating cells of a ybaN ytrI double mutant and of a ybaN ytrHI triple mutant exhibited a pronounced loss of dipicolinic acid (DPA) between hours 8 and 24 of sporulation, in contrast to the constant levels seen for the wild type. An analysis of the spore cortex peptidoglycans of the ybaN ytrI and ybaN ytrHI mutants showed striking decreases in the levels of total muramic acid by hour 24 of sporulation. These data, along with the loss of DPA in the mutants, suggest that the developing spores were unstable and that the cortex underwent degradation late in sporulation. The existence of otherwise hidden sporulation pathways indicates that functional redundancy may mask the role of hitherto unrecognized sporulation genes. PMID- 15547283 TI - Initial Phases of biofilm formation in Shewanella oneidensis MR-1. AB - Shewanella oneidensis MR-1 is a facultative Fe(III)- and Mn(IV)-reducing microorganism and serves as a model for studying microbially induced dissolution of Fe or Mn oxide minerals as well as biogeochemical cycles. In soil and sediment environments, S. oneidensis biofilms form on mineral surfaces and are critical for mediating the metabolic interaction between this microbe and insoluble metal oxide phases. In order to develop an understanding of the molecular basis of biofilm formation, we investigated S. oneidensis biofilms developing on glass surfaces in a hydrodynamic flow chamber system. After initial attachment, growth of microcolonies and lateral spreading of biofilm cells on the surface occurred simultaneously within the first 24 h. Once surface coverage was almost complete, biofilm development proceeded with extensive vertical growth, resulting in formation of towering structures giving rise to pronounced three-dimensional architecture. Biofilm development was found to be dependent on the nutrient conditions, suggesting a metabolic control. In global transposon mutagenesis, 173 insertion mutants out of 15,000 mutants screened were identified carrying defects in initial attachment and/or early stages in biofilm formation. Seventy-one of those mutants exhibited a nonswimming phenotype, suggesting a role of swimming motility or motility elements in biofilm formation. Disruption mutations in motility genes (flhB, fliK, and pomA), however, did not affect initial attachment but affected progression of biofilm development into pronounced three-dimensional architecture. In contrast, mutants defective in mannose-sensitive hemagglutinin type IV pilus biosynthesis and in pilus retraction (pilT) showed severe defects in adhesion to abiotic surfaces and biofilm formation, respectively. The results provide a basis for understanding microbe-mineral interactions in natural environments. PMID- 15547284 TI - Crystal structure of the 65-kilodalton heat shock protein, chaperonin 60.2, of Mycobacterium tuberculosis. AB - Chaperonin 60s are a ubiquitous class of proteins that promote folding and assembly of other cellular polypeptides in an ATP-dependent manner. The oligomeric state of chaperonin 60s has been shown to be crucial to their role as molecular chaperones. Chaperonin 60s are also known to be important stimulators of the immune system. Mycobacterium tuberculosis possesses a duplicate set of chaperonin 60s, both of which have been shown to be potent cytokine stimulators. The M. tuberculosis chaperonin 60s are present in the extracellular milieu at concentrations that are extremely low for the formation of an oligomer. Here we present the crystal structure of one of the chaperonin 60s of M. tuberculosis, also called Hsp65 or chaperonin 60.2, at 3.2-A resolution. We were able to crystallize the protein in its dimeric state. The unusual dimerization of the protein leads to exposure of certain hydrophobic patches on the surface of the protein, and we hypothesize that this might have relevance in binding to immunogenic peptides, as it does in the eukaryotic homologs. PMID- 15547285 TI - Transferable antibiotic resistance elements in Haemophilus influenzae share a common evolutionary origin with a diverse family of syntenic genomic islands. AB - Transferable antibiotic resistance in Haemophilus influenzae was first detected in the early 1970s. After this, resistance spread rapidly worldwide and was shown to be transferred by a large 40- to 60-kb conjugative element. Bioinformatics analysis of the complete sequence of a typical H. influenzae conjugative resistance element, ICEHin1056, revealed the shared evolutionary origin of this element. ICEHin1056 has homology to 20 contiguous sequences in the National Center for Biotechnology Information database. Systematic comparison of these homologous sequences resulted in identification of a conserved syntenic genomic island consisting of up to 33 core genes in 16 beta- and gamma-Proteobacteria. These diverse genomic islands shared a common evolutionary origin, insert into tRNA genes, and have diverged widely, with G+C contents ranging from 40 to 70% and amino acid homologies as low as 20 to 25% for shared core genes. These core genes are likely to account for the conjugative transfer of the genomic islands and may even encode autonomous replication. Accessory gene clusters were nestled among the core genes and encode the following diverse major attributes: antibiotic, metal, and antiseptic resistance; degradation of chemicals; type IV secretion systems; two-component signaling systems; Vi antigen capsule synthesis; toxin production; and a wide range of metabolic functions. These related genomic islands include the following well-characterized structures: SPI-7, found in Salmonella enterica serovar Typhi; PAP1 or pKLC102, found in Pseudomonas aeruginosa; and the clc element, found in Pseudomonas sp. strain B13. This is the first report of a diverse family of related syntenic genomic islands with a deep evolutionary origin, and our findings challenge the view that genomic islands consist only of independently evolving modules. PMID- 15547286 TI - Regulation of iron transport in Streptococcus pneumoniae by RitR, an orphan response regulator. AB - RitR (formerly RR489) is an orphan two-component signal transduction response regulator in Streptococcus pneumoniae that has been shown to be required for lung pathogenicity. In the present study, by using the rough strain R800, inactivation of the orphan response regulator gene ritR by allele replacement reduced pathogenicity in a cyclophosphamide-treated mouse lung model but not in a thigh model, suggesting a role for RitR in regulation of tissue-specific virulence factors. Analysis of changes in genome-wide transcript mRNA levels associated with the inactivation of ritR compared to wild-type cells was performed by the use of high-density DNA microarrays. Genes with a change in transcript abundance associated with inactivation of ritR included piuB, encoding an Fe permease subunit, and piuA, encoding an Fe carrier-binding protein. In addition, a dpr ortholog, encoding an H(2)O(2) resistance protein that has been shown to reduce synthesis of reactive oxygen intermediates, was activated in the wild-type (ritR(+)) strain. Microarray experiments suggested that RitR represses Fe uptake in vitro by negatively regulating the Piu hemin-iron transport system. Footprinting experiments confirmed site-specific DNA-binding activity for RitR and identified three binding sites that partly overlap the +1 site for transcription initiation upstream of piuB. Transcripts belonging to other gene categories found to be differentially expressed in our array studies include those associated with (i) H(2)O(2) resistance, (ii) repair of DNA damage, (iii) sugar transport and capsule biosynthesis, and (iv) two-component signal transduction elements. These observations suggest that RitR is an important response regulator whose primary role is to maintain iron homeostasis in S. pneumoniae. The name ritR (repressor of iron transport) for the orphan response regulator gene, rr489, is proposed. PMID- 15547287 TI - Vibrio cholerae strains with mutations in an atypical type I secretion system accumulate RTX toxin intracellularly. AB - This study shows that the Vibrio cholerae RTX toxin is secreted by a four component type I secretion system (TISS) encoded by rtxB, rtxD, rtxE, and tolC. ATP-binding site mutations in both RtxB and RtxE blocked secretion, demonstrating that this atypical TISS requires two transport ATPases that may function as a heterodimer. PMID- 15547288 TI - The malic enzyme is required for optimal photoautotrophic growth of Synechocystis sp. strain PCC 6803 under continuous light but not under a diurnal light regimen. AB - A mutation was recovered in the slr0721 gene, which encodes the decarboxylating NADP(+)-dependent malic enzyme in the cyanobacterium Synechocystis sp. strain PCC 6803, yielding the mutant 3WEZ. Under continuous light, 3WEZ exhibits poor photoautotrophic growth while growing photoheterotrophically on glucose at rates nearly indistinguishable from wild-type rates. Interestingly, under diurnal light conditions (12 h of light and 12 h of dark), normal photoautotrophic growth of the mutant is completely restored. PMID- 15547289 TI - The chemical chaperone proline relieves the thermosensitivity of a dnaK deletion mutant at 42 degrees C. AB - Since, like other osmolytes, proline can act as a protein stabilizer, we investigated the thermoprotectant properties of proline in vitro and in vivo. In vivo, elevated proline pools in Escherichia coli (obtained by altering the feedback inhibition by proline of gamma-glutamylkinase, the first enzyme of the proline biosynthesis pathway) restore the viability of a dnaK-deficient mutant at 42 degrees C, suggesting that proline can act as a thermoprotectant for E. coli cells. Furthermore, analysis of aggregated proteins in the dnaK-deficient strain at 42 degrees C by two-dimensional gel electrophoresis shows that high proline pools reduce the protein aggregation defect of the dnaK-deficient strain. In vitro, like other "chemical chaperones," and like the DnaK chaperone, proline protects citrate synthase against thermodenaturation and stimulates citrate synthase renaturation after urea denaturation. These results show that a protein aggregation defect can be compensated for by a single mutation in an amino acid biosynthetic pathway and that an ubiquitously producible chemical chaperone can compensate for a defect in one of the major chaperones involved in protein folding and aggregation. PMID- 15547290 TI - A self-splicing group I intron in DNA polymerase genes of T7-like bacteriophages. AB - Group I introns are inserted into genes of a wide variety of bacteriophages of gram-positive bacteria. However, among the phages of enteric and other gram negative proteobacteria, introns have been encountered only in phage T4 and several of its close relatives. Here we report the insertion of a self-splicing group I intron in the coding sequence of the DNA polymerase genes of PhiI and W31, phages that are closely related to T7. The introns belong to subgroup IA2 and both contain an open reading frame, inserted into structural element P6a, encoding a protein belonging to the HNH family of homing endonucleases. The introns splice efficiently in vivo and self-splice in vitro under mild conditions of ionic strength and temperature. We conclude that there is no barrier for maintenance of group I introns in phages of proteobacteria. PMID- 15547291 TI - Type II isopentenyl diphosphate isomerase from Synechocystis sp. strain PCC 6803. AB - Open reading frame sll1556 in the cyanobacterium Synechocystis sp. strain 6803 encodes a putative type II isopentenyl diphosphate (IPP) isomerase. The His(6) tagged protein was produced in Escherichia coli and purified by Ni(2+) chromatography. The homotetrameric enzyme required NADPH, flavin mononucleotide, and Mg(2+) for activity; K(m)(IPP) was 52 microM, and k(cat)(IPP) was 0.23 s(-1). PMID- 15547292 TI - Thematic review series: the immune system and atherogenesis. Immune function in atherogenesis. AB - In this overview to a new thematic series on the immune system and atherogenesis, I provide a very brief summary of current conceptions of atherogenesis, of the innate and adaptive immune systems, and of the participation of the latter in atherogenesis, with particular emphasis on studies of the involvement of the immune system in atherosclerosis reported in the last 2 years. This is followed by a short outline of the eight reviews that will make up this thematic series. The overview is concluded with some caveats that should be considered in the analysis of atherosclerosis in experimental animals. PMID- 15547293 TI - Thematic review series: the pathogenesis of atherosclerosis. An interpretive history of the cholesterol controversy: part II: the early evidence linking hypercholesterolemia to coronary disease in humans. AB - The first in this series of historical reviews dealt with the pioneering animal model work of Anitschkow, implicating blood cholesterol in the pathogenesis of atherosclerosis, and the pivotally important work of Gofman, providing evidence that lipoprotein-bound cholesterol was a major factor in the human disease. This second installment reviews the early lines of evidence linking hypercholesterolemia in humans to the progression of atherosclerosis and the risk of coronary heart disease. The argument is made that by 1970, the evidence was already strong enough to justify intervention to lower blood cholesterol levels if all the available lines of evidence had been taken into account. Yet, it would be almost two decades before lowering blood cholesterol levels became a national public health goal. Some of the reasons the "cholesterol controversy" continued in the face of powerful evidence supporting intervention are discussed. PMID- 15547294 TI - Transcriptional regulation of human microsomal triglyceride transfer protein by hepatocyte nuclear factor-4alpha. AB - Microsomal triglyceride transfer protein (MTP) catalyzes the assembly of triglyceride (TG)-rich apolipoprotein B-containing liver (e.g., VLDL) and intestinal (e.g., chylomicron) lipoproteins. The human MTP gene promoter is reported here to associate in vivo with endogenous hepatocyte nuclear factor 4alpha (HNF-4alpha) and to be transactivated or transsuppressed by overexpressed or by dominant negative HNF-4alpha, respectively. Human MTP (hMTP) transactivation by HNF-4alpha is accounted for by the concerted activity of distal (-83/-70) and proximal (-50/-38) direct repeat 1 elements of the hMTP promoter that bind HNF-4alpha. Transactivation by HNF-4alpha is specifically antagonized by chicken ovalbumin upstream promoter. Transcriptional activation of hMTP by HNF-4alpha is mediated by HNF-4alpha domains engaged in ligand binding and ligand-driven transactivation and is further complemented by HNF-4alpha/HNF 1alpha synergism that involves the HNF-4alpha activation function 1 (AF-1) domain. hMTP transactivation by HNF-4alpha is specifically inhibited by beta,beta tetramethyl-hexadecanedioic acid acting as an HNF-4alpha antagonist ligand. hMTP transactivation by HNF-4alpha may account for the activation or inhibition of MTP expression and the production of TG-rich lipoproteins by agonist (e.g., saturated fatty acids) or antagonist [e.g., (n-3) PUFA, hypolipidemic fibrates, or Methyl substituted dicarboxylic acid (Medica) compounds] HNF-4alpha ligands. PMID- 15547295 TI - Intestinal fatty acid binding protein and microsomal triglyceride transfer protein polymorphisms in French-Canadian youth. AB - Growing evidence suggests an association between lipid abnormalities and fatty acid binding protein (FABP) and microsomal triglyceride transfer protein (MTP) gene variants. Our objectives were to determine whether Ala54Thr FABP2 and G-493T MTP polymorphisms are associated with increased risks of insulin resistance syndrome (IRS) in youth and/or modify the expression of accompanying dyslipidemia. Our study of 1,742 French-Canadians aged 9, 13, and 16 years did not provide evidence of a potential predisposition to IRS related to either FABP2 or MTP genotypes. However, we observed a heterogeneity of the FABP2 effect by IRS status on total cholesterol (TC), low density lipoprotein-cholesterol (LDL-C), and apolipoprotein B (apoB) concentrations (P for interaction=0.045, 0.018, and 0.017, respectively). Among the metabolic components of IRS, only triglyceride (TG) displayed an interaction with FABP2 polymorphism: compared with Thr/Ala and Ala/Ala, the Thr/Thr genotype was associated with a steeper increase in TC, LDL C, and apoB parallel to TG concentrations (P <0.001). IRS did not modify the associations between the MTP polymorphism and any of the biochemical parameters. Our study suggests that the effects of FABP2 allelic variations on lipid traits are context dependent, indicating that this variant may play an important role in cardiovascular pathogenesis in the presence of IRS or hypertriglyceridemia. PMID- 15547296 TI - Interleukin-10 enhances the oxidized LDL-induced foam cell formation of macrophages by antiapoptotic mechanisms. AB - Interleukin (IL)-10 may have a therapeutic potential in atherosclerosis, but its mechanisms of action have not been clarified. Foam cell formation is a key event in atherogenesis, and apoptosis of these lipid-laden cells may promote plaque destabilization. We sought to explore whether IL-10 could have plaque-stabilizing properties in acute coronary syndromes (ACS). We studied the effect of IL-10 on oxidized low density lipoprotein (oxLDL)-stimulated THP-1 cells and monocyte derived macrophages from ACS patients and healthy controls using different experimental approaches. Our main findings were: i) IL-10 enhances lipid accumulation in oxLDL-stimulated THP-1 macrophages, at least partly by counteracting oxLDL-induced apoptosis; ii) This antiapoptotic effect of IL-10 involves increased expression of the antiapoptotic genes Bfl-1 and Mcl-1, accompanied by protective effects on mitochondria function; iii) By silencing Bfl 1 and Mcl-1 genes using siRNAs, we were able to abolish this IL-10-mediated effect on lipid accumulation; iv) IL-10 also induced lipid accumulation in oxLDL stimulated macrophages from patients with ACS, but not in macrophages from healthy controls; v) In ACS patients, this enhancing effect of IL-10 on lipid accumulation was accompanied by enhanced Mcl-1 expression. No such antiapoptotic effect was seen in macrophages from healthy controls. These findings suggest a new mechanism for the effect of IL-10 in atherosclerosis, possibly contributing to plaque stabilization. PMID- 15547297 TI - Identification and analysis of products formed from phospholipids in the free radical oxidation of human low density lipoproteins. AB - Phospholipids reside in the surface layer of LDLs and constitute approximately 20 25% of the particle by weight. We report a study of the primary products generated from the most abundant molecular species of phosphatidylcholines present in LDL during in vitro free radical oxidations. The 13-hydroperoxides of 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine (PLPC) and 1-stearoyl-2 linoleoyl-sn-glycero-phosphocholine (SLPC) and the 15-hydroperoxides of 1 palmitoyl-2-arachidonyl-sn-glycero-3-phosphocholine (PAPC) and 1-stearoyl-2 arachidonoyl-sn-glycero-phosphocholine (SAPC) were found to increase in a time dependent manner and in significant amounts even in the presence of alpha tocopherol. Phospholipid alcohols also formed during the course of the oxidations. Early in the LDL oxidations, while alpha-tocopherol was still present, the thermodynamically favored trans,trans products of PLPC and SLPC were found to form in significantly larger quantities than those formed from cholesteryl linoleate. Additionally, quantities of PAPC 11-hydroperoxide (11-OOH) decreased over time relative to PAPC 15-OOH, even while alpha-tocopherol was still present in the oxidation, presumably as a result of further oxidation of PAPC 11-OOH to form cyclic peroxide oxidation products. These results suggest that alpha-tocopherol is more closely associated with the inner cholesteryl ester rich hydrophobic core of an LDL particle and is not as effective as an antioxidant in the outer phospholipid layer as it is in the lipid core. PMID- 15547298 TI - The effect of sterol regulatory element-binding protein 2 polymorphism on the serum lipid in northern Chinese subjects. AB - Sterol regulatory element-binding protein 2 (SREBP-2) is an important nuclear transcription factor in the regulation of cellular cholesterol metabolism. To determine allele frequency of the 1784G>C polymorphism at the SREBP-2 locus and investigate the relationship between this polymorphism and serum lipid levels in Chinese people, we selected 486 individuals (118 men and 368 women) from the Xicheng District of Beijing. The subjects were divided into four groups: hypercholesterolemic subjects, hypertriglyceridemic subjects, combined hyperlipidemic subjects, and normal subjects. Serum lipid profiles were measured in all subjects, and 1784G>C was analyzed using polymerase chain reaction restriction fragment length polymorphism analysis. There was no significant difference in genotype frequencies or allele frequencies of this polymorphism between the hyperlipidemic and control groups. The serum total cholesterol (TC) and LDL cholesterol (LDL-C) levels of the individuals carrying the C allele were higher than the noncarriers in both males and females in the hypercholesterolemic group, but statistical significance was only observed in females. The results of this study indicate that the SREBP-2 polymorphism is related to elevated concentrations of serum TC and LDL-C in hypercholesterolemic subjects. Further work is necessary to confirm the role of 1784G>C in the development of hyperlipidemia. PMID- 15547299 TI - Fatty acid flip-flop and proton transport determined by short-circuit current in planar bilayers. AB - The effect of palmitic acid (PA) and oleic acid (OA) on electrical parameters of planar membranes was studied. We found a substantial difference between the effects of PA and OA on proton transfer. PA induced a small increase in conductance, requiring a new technique for estimating proton-mediated currents across low-conductance planar bilayers in which an electrometer is used to measure the transmembrane current under virtual short circuit (SCC). Open-circuit voltage and SCC were used to determine proton and leak conductances. OA caused a marked increase in membrane conductance, allowing the use of a voltage-clamp technique. From SCC data, we were able to estimate the flip-flop rate constants for palmitate (1 x 10(-6) s(-1)) and oleate (49 x 10(-6) s(-1)) anions. Cholesterol, included in the membrane-forming solution, decreased importantly the leak conductance both in membranes unmodified by FA and in membranes modified by PA added to the bath. PMID- 15547300 TI - Impact of adiponectin gene polymorphisms on plasma lipoprotein and adiponectin concentrations of viscerally obese men. AB - The aim of this study was first to examine the relationships between adiponectin gene (Apm1) polymorphisms and anthropometric indices as well as plasma adiponectin and lipoprotein/lipid levels, and then to investigate whether the presence of visceral obesity or insulin resistance may modulate the impact of these polymorphisms on metabolic risk variables. Molecular screening of the Apm1 gene was achieved, and a sample of 270 unrelated men recruited from the greater Quebec City area and selected to cover a wide range of body fatness values was genotyped. Sequencing of the Apm1 gene revealed two previously reported polymorphisms (c.45T>G and c.276G>T) as well as two newly identified genetic variations (-13752delT and -13702G>C). Carriers of the c.276T allele had higher LDL-cholesterol and lower HDL-triglyceride concentrations than did 276G/G homozygotes (P=0.02 and P=0.01, respectively). Carriers of the c.45G allele exhibited higher plasma adiponectin concentrations than did 45T/T homozygotes (P=0.04). After dividing each genotype group into subgroups for visceral AT, homozygotes for the normal allele at position -13752delT, carriers of the c.45G allele, and carriers of the c.276T allele had similar total apolipoprotein B (apoB) concentrations, whether they were viscerally obese or not. These results suggest that some Apm1 gene polymorphisms influence plasma adiponectin concentrations and lipoprotein/lipid levels. In addition, the impact of these polymorphisms is modulated by the presence of visceral obesity. PMID- 15547301 TI - Real-time quantification of fatty acid uptake using a novel fluorescence assay. AB - Uptake of nonesterified long-chain fatty acids (LCFAs) into many cell types and organs such as liver, heart, intestine, and skeletal muscle occurs primarily through a saturable, protein-mediated mechanism. Membrane proteins that increase the uptake of LCFAs, such as FAT/CD36 and fatty acid transport proteins, represent significant therapeutic targets for the treatment of metabolic disorders, including type 2 diabetes. However, currently available methods for the quantification of LCFA uptake neither allow for real-time measurements of uptake kinetics nor are ideally suited for the development of LCFA uptake inhibitors in high-throughput screens. To address both problems, we developed a LCFA uptake assay using a fluorescently labeled fatty acid and a nontoxic cell impermeable quenching agent that allows fatty acid transport to be measured in real time using fluorescence plate readers or standard fluorescence microscopy. With this assay, we faithfully reproduced known differentiation- and hormone induced changes in LCFA uptake by 3T3-L1 cells and determined LCFA uptake kinetics with previously unobtainable temporal resolution. Applications of this novel assay should facilitate new insights into the biology of fatty acid uptake and provide new means for obesity-related drug discovery. PMID- 15547302 TI - Serum sphingomyelin levels are related to the clearance of postprandial remnant like particles. AB - It is known that sphingomyelin (SM) content is higher in apolipoprotein B containing particles (BLps) than in high density lipoproteins and that BLp levels, including chylomicrons and their remnant particles, are positively related to atherosclerosis. To evaluate the relationship between serum SM and postprandial remnant particle levels, we determined SM, triglyceride (TG), and cholesterol levels in serum and in remnant-like particles (RLPs) before and 3, 5, 7, and 10 h after a high-fat meal in 31 healthy subjects. We found that serum SM, like serum TG, was increased to its maximum 3 h after fat loading and then gradually decreased to basal levels after 10 h. More important, we determined that SM and TG levels in RLPs were parallel. Serum SM was positively correlated with serum TG (P <0.001), RLP SM (P <0.001), RLP TG (P <0.001), and RLP cholesterol (P <0.001) levels. It is our conclusion that serum SM is a marker for the clearance of RLPs. PMID- 15547303 TI - [Lithuanian people are waiting for the medical profession firm word and action]. PMID- 15547304 TI - [Thrombolysis during cardiopulmonary resuscitation]. AB - It has been estimated that 50-70% of cardiac arrests are caused by acute myocardial infarction or fulminant pulmonary embolism. Thrombolysis could be the treatment of choice during cardiopulmonary resuscitation given its ability to treat the underlying cause of the condition. Traditionally thrombolysis during cardiopulmonary resuscitation has been contraindicated because of risk of life threatening bleeding complications. Early clinical experience and results of several trials suggest that the risk of bleeding is lower and the overall clinical benefit is greater than previously thought. Imbalance between coagulation and fibrinolysis develops during cardiopulmonary resuscitation, followed by disseminated clotting in small vessels. This condition causes inadequate reperfusion of the brain. Experimental studies demonstrate that besides dissolving coronary thrombus or pulmonary emboli, thrombolytic therapy improves microcirculatory flow resulting in better reperfusion of the brain which translates into better neurological outcome after resuscitation. Should the currently ongoing "Thrombolysis In Cardiac Arrest - TROICA Study" confirm the results of the earlier trials, thrombolytic therapy may soon become part of the resuscitation guidelines. PMID- 15547305 TI - [The impact of preoperative stoma siting and stoma care education on patient's quality of life]. AB - The aim of study was to assess if preoperative stoma selection and adequate patient's teaching can affect the postoperative patient's quality of life. The study was performed in two university hospitals of Lithuania: Kaunas University of Medicine Hospital and Kaunas Oncology Hospital. Patients were divided into three groups. Patients were asked to answer the questionnaire the day before the stoma creation operation and two months after the operation. Questionnaires EORTC QLQ-C30, EORTC QLQ-CR38 and 10 supplementary questions were used. The results of the study show that following the stoma operation, when compared with preoperative results, general quality of life did not change significantly in groups I and II. Patients who received adequate education and preoperative stoma siting had better emotional functioning and less gastrointestinal problems. The financial problems of the patients in group I were significantly less than in the control group. Patients who received the adequate teaching without preoperative stoma selection experience better sexual satisfaction compared with control group. Stoma related problems were less in group I and II when comparing with the control group. The quality of the patients' teaching, adequacy and comfort of stoma site and satisfaction with the medical staff were significantly better in the group I and group II when compared to control group. Moreover, these results were significantly higher in the group I than in group II. CONCLUSIONS. The teaching the patients preoperatively and postoperative proceeding helps them to gain better experience in self stoma care hence reducing the psychological, physical, emotional, social and sexual problems. PMID- 15547306 TI - [Laparoscopic repair of perforated duodenal ulcer: early postoperative results and risk factors]. AB - BACKGROUND: Clear patient selection criteria and indications for laparoscopic repair of perforated duodenal ulcers are still of relevance. The purpose of our paper is to describe the early outcome results after this operation and to define the risk factors influencing the genesis of postoperative morbidity. PATIENTS AND METHODS: Fifty-one patients were operated on laparoscopically between October 1996 and October 2003 for perforated peptic ulcers. Out of them, 47 patients with perforated duodenal ulcers entered the final retrospective analysis. Twenty variables were identified, including the duration of acute symptoms before the operation, shock, underlying medical illness, ulcer size, age, Boey score and the predictive value of these variables for morbidity, conversion rates and hospital stay. The univariate data analysis was originally done using the Fisher exact test, t test, Mann Whitney, ANOVA and F tests. The data was reevaluated using multifactorial analysis with logistic and linear regression tests. RESULTS: Patient's age was 32.0+/-12.4 years. Duration of perforation was 8.0+/-10.8 hours. Shock was diagnosed in one patient (2.1%). High surgical risk according to ASA (III-IV) was estimated in 2 patients (4.3%). Laparoscopic duodenography was completed in 36 patients (76.6%). The other 11 patients (23.4%) underwent a conversion to open repair. Seven patients (19.4%) had postoperative complications. Suture leakage was confirmed in 4 patients (11.1%), and other abdominal complications were observed in 3 patients (8.3%). Pneumonia and pneumothorax were diagnosed in 3 patients. Hospital stay was 7.9+/-5.8 (4-45) days. There was no mortality. Ulcer perforation size >4-10 mm is the only significant risk factor influencing the conversion rate. An increase in the suture leakage rate was most significant with delayed presentation of >or=10 hours (p<0.0001). This risk factor influences both the postoperative pneumonia rate (>or=10 hrs., p=0.026) and hospital stay (p<0.05). CONCLUSION: Size of duodenal ulcer perforation and duration of ulcer perforation symptoms were found to be risk factors influencing the rates of conversion to open repair and genesis of postoperative morbidity. PMID- 15547307 TI - [Clinical results after surgeries of intraarticular distal radius fractures]. AB - The aim of the study was to evaluate treatment results of intraarticular distal radius fractures using the combined method and to compare it to other methods. MATERIAL AND METHODS: Between 2000 and 2003 we evaluated clinical results of 105 patients with intraarticular fracture of distal radius. Thirty-five patients underwent the repair of intraarticular fracture according to combined method (using Kirschner wires and external fixator), 30 patients - only with Kirschner wires and 40 patients - only with external fixator. Results were evaluated clinically and rentgenologically. The evaluation was made by scoring system according to J. J. Gartland and C. W. Werley (modified by A. Sarmiento). X-ray pictures were evaluated every 2 weeks till the 6(th) week ant after 6 months. RESULTS: Using combined method, treatment results were good and very good for 28 patients, and satisfactory for 7 patients. In case when Kirschner wires were used, good and very good results were found in 17 patients, and satisfactory - in 13 ones. When only external fixators were used, good and very good results were observed in 26 patients and satisfactory - in 14 patients. There was statistically significant difference between the groups (p<0.05). CONCLUSIONS: Better clinical treatment results of intraarticular distal radius fractures were achieved when combined method (using Kirschner wires and external fixator) was applied. Using other methods of treatment we observed more cases of secondary dislocations. PMID- 15547308 TI - Evaluation of clinical factors and treatment results in patients with advanced pancreatic cancer. AB - OBJECTIVE: The aim of the study was to assess the benefit of treatment modalities on the survival in patients with advanced pancreatic cancer and clinical factors affecting treatment efficacy and survival. MATERIAL AND METHODS: One hundred eleven patients with advanced pancreatic cancer were analyzed retrospectively. Patients were grouped by treatment method, clinical stage, and Karnofsky Performance Index. Fifty-three patients were diagnosed with locally advanced disease and 58 with metastatic pancreatic cancer. Thirty-three patients at the time of diagnosis had Karnofsky Performance Index higher than 70, and in 78 patients it was 70 or lower. Fourteen patients were treated by concomitant chemoradiotherapy with gemcitabine or 5-fluorouracil, 25 - with gemcitabine only, and 72 patients underwent surgical palliation or observation alone. RESULTS: Patients treated with gemcitabine alone survived for 9.5 months, p<0.001. Overall median survival of patients treated with concomitant chemoradiation was 8.5 months. Comparison of survival results between groups of patients treated with gemcitabine alone and the patients who have received radiation therapy with 5 fluorouracil (median survival 6.4 months) or gemcitabine (median survival - 8.8 months) revealed no difference. Median survival after surgical palliation or observation was 1.9 months. Patients diagnosed with locally advanced pancreatic cancer and patients with Karnofsky Performance Index higher than 70 at diagnosis lived statistically longer than patients diagnosed with metastatic disease or Karnofsky Performance Index of 70 or lower. CONCLUSIONS: Patients diagnosed with locally advanced pancreatic cancer, better performance status at diagnosis and treated with monochemotherapy with gemcitabine or combination of gemcitabine or 5 fluorouracil with radiation survived longer, than patients diagnosed with metastatic disease, patients of worse functional status and treated by palliative methods only. PMID- 15547309 TI - Role of the skin patch test in diagnosing food allergy in children with atopic dermatitis. AB - The aim of the study was to determine peculiarities of food allergy in children with atopic dermatitis and to evaluate the significance of skin patch test in determining the main food allergens. METHODS: One hundred and eight children (57 boys and 51 girls) with atopic dermatitis were examined. Atopic dermatitis was diagnosed by standard diagnostic criteria, severity of the progress of the disease was determined using SCORAD index and the amount of total IgE in blood, skin prick and patch tests with the main food allergens were performed. RESULTS: The age of the patients varied from 6 months to 16 years, however, almost half (41%) of them were toddlers (1-3 years old). Mild form of atopic dermatitis was dominating (52%). Analysis of the total IgE amount in blood showed different degree of sensitivity of the children tested. Normal amount of the total IgE in blood was found in 73.1% of children with atopic dermatitis, and the increased total IgE amount was found only in 26.9% of children. Positive skin prick test with the standard and the most common food allergens was found only in 4.63% of children with atopic dermatitis, while the positive skin patch test with 25 food allergens was found in 68.5% of children. Depending on the type of the allergic reaction, immediate type reaction dominated only in 10.3% of children with atopic dermatitis, while the delayed type allergic reactions were characteristic to food allergies in 48.3% of children with atopic dermatitis. Food allergy was not found in one fifth of children with atopic dermatitis. CONCLUSIONS: Skin patch test is an informative and reliable diagnostic test in evaluating the delayed type allergic reactions. In about half of the tested persons with atopic dermatitis, food allergy appeared in delayed type allergic reactions. Therefore it is very important to do the skin patch test for toddlers and pre-school age children. The most common allergens found with the help of skin patch test are soy, milk, peanuts, carrot, egg whites, wheat, and corn. PMID- 15547310 TI - [Women's opinion on hormone pills and the main sources of information]. AB - The aim of this paper was to analyze and to evaluate women's opinion on hormone pills and the influence on hormone pill usage and reveal the main sources of information. MATERIAL AND METHODS: Women who came for pregnancy termination (Group A) and women who came for consultation on further usage of contraception (Group K) have been interrogated. Interrogation according the special questionnaire has been performed at four consultation centers and three gynecologic clinics of Kaunas city. It was find out that due to the wrong information 25.1% of women in group A and 35% of women in group K thought that hormone pills cause irregular bleeding (p=0.01). Another myth - the possibility to become pregnant when taking pills - was indicated by 17% of women in group A and 32.4% of women in group K (p=0.01). Nine percent in group A and 12% in group K were confident that when taking hormone pills for some period of time one will not be able to become pregnant in the future. Seventy three percent of group A women and 80% of group K women indicated that the use of hormone pills increases the body weight (p=0.01). This research showed that the women that used hormone pills were properly informed about their treatment and side effects. Fifty four percent of group A women and 36% of group K women indicated that hormone pills were not suitable in case of several diseases (p=0.01). Eighty three percent of group A women and 71% of group K knew that hormone pills did not protect from cervical cancer (p=0.01). This information was mainly obtained from medical workers. The main reason why women of fertile age who took part in our research used little of hormone pills was false information received from insufficiently skilled persons and accordingly the negative opinion about hormone pills. CONCLUSIONS: Women's opinion on hormone pills is a very important factor for its usage. This opinion depends on sources of information, whether women received correct or false information on hormone pills. The most reliable source of information indicated by investigated women was medical personnel. PMID- 15547311 TI - [Effect of pyruvate on the dependence of electromechanical activity on stimulation frequency in human myocardium]. AB - The aim of the study was to determine the effect of pyruvate, a substrate of energetic metabolism, on the dependence of electromechanical activity on stimulation frequency in human myocardium. The experiments were performed on human ventricle myocardium from patients undergoing cardiac corrective open heart surgery at Clinic of Cardiac Surgery in Kaunas University of Medicine by using standard method of registration of myocardium electromechanical activity. The stimulation frequency of myocardial strips were 0.2, 0.5, 1.0, 1.5, 2.0, 2.5 and 3 Hz, duration of pulses -2-5 ms. In control, i.e. at perfusion of mycoardial strips by Tyrode solution and stimulation frequency 1 Hz, an average of contraction force (P) was 0.94+/-0.18 mN (n=12), half time of relaxation (t(a)) - 178.88+/-9.31 ms (n=12), and action potential duration measured at 50% repolarization level (VP(50)) - 294.75+/-27.95 ms (n=9). Pyruvate (10 mM) increased P to 159.9+/-13.4% (p<0.00), t(a)- 106.9+/-2.2% (n=12) (p<0.001), and VP(50)- 111.1+/-7.4% (n=5) (p<0.05), as compared to control. In the absence of pyruvate values of P, t(a) and VP(50) of myocardial strips were higher at 0.2 Hz and 0.5 Hz and lower at 3 Hz compared with those at 1 Hz of stimulation frequency. Although the dependence of these parameters on stimulation frequency in the presence of pyruvate (10 mM) was not altered but their values were persistently increased at all stimulation frequencies. The present data indicate that pyruvate is an effective inotropic agent, which can improve contractility function in wide-ranging diapason of stimulation frequency in failing human heart. PMID- 15547312 TI - [Effect of hypodynamic stress on the adrenaline-induced contraction of the smooth muscles of the thoracic aorta]. AB - Aim of the study was to investigate the influence of hypodynamic stress on the contractile responses of the smooth muscles to adrenaline. MATERIAL AND METHODS: Hypodynamic stress was induced by fixation of Chinchilla rabbits (n=8) in the metal hutches for 48 days (B. M. Fiodorov method). Rabbits in the control group (n=8) were kept in ordinary conditions of vivarium for the same period of time. Isolated preparations of the thoracic aorta were obtained from both groups of the rabbits. The contractile responses of the thoracic aorta preparations to 10( 7)mol/l and 10(-6)mol/l of adrenaline were registered in vitro by micromechanographic device under isometric conditions. RESULTS: The contractile responses of the thoracic aorta preparations under the influence of adrenaline at concentration 10(-7)mol/l were not significantly higher (p>0.05) and at concentration 10(-6)mol/l they were significantly higher (p=0.021) in rabbits after hypodynamic stress than in control ones. CONCLUSION: Long-term hypodynamic stress in rabbits leads to the significantly increased adrenaline-induced contraction of the smooth muscles of the thoracic aorta. PMID- 15547313 TI - The patient role in decision-making in Lithuanian health care. AB - The objective of the study is to explore the attitudes of Lithuanian citizens to health care system and to evaluate their social role in decision-making about their health care. MATERIAL AND METHODS: The national cross-sectional survey was conducted March 1-7, 2004. Multi-stage random sampling across the country included adult Lithuanian residents, interviewed in their home (n=1007). The response rate was 56.5%. The attitudes of citizens were evaluated using an anonymous questionnaire, originally developed and adopted with the reference to experience of previously conducted studies. RESULTS: Majority of polled women and men (84.6% and 72.6%) recognized health as very important value in their life and as the most important among other social values. The overall mean of trust in health care system - 41.3%, trust in physicians - 69.9%, implementation of right to health care - 48.9%, concern about health care - 96.5% and patient impact in health care decisions - 19.1%. Nearly half of respondents (47.4%) prefer informative general practitioner-patient interaction model, which is realized in most cases (58.8%). Partnership (shared decision-making) as interaction model is expected by 37.2% of polled respondents and realized in every sixth case in primary care. CONCLUSIONS: Health as value is the most preferred among Lithuanian citizens. Younger and more educated people trusted less in health care system, but are more motivated to play an active role in health care decision-making. The informative model of doctor-patient is dominant, while partnership is not so exposed as being expected by patients. PMID- 15547314 TI - Alcohol consumption in Lithuanian school-aged children during 1994-2002. AB - Alcohol consumption becomes an important social and health problem among youth in many countries. Analysis of data on alcohol consumption behavior from World Health Organization Cross-National Study on Health Behavior in School-aged Children (HBSC) is presented in the article. This survey is carried out in majority of European countries including Lithuania. Aim of this study was to analyze features and trends of alcohol consumption among representative sample of 11, 13 and 15-year-old school children and to compare results with international data. METHODS: Patterns and trends of alcohol consumption were analyzed in three cross sectional questionnaire surveys of the representative sample of Lithuanian secondary school student's aged 11, 13 and 15 years in 1994, 1998 and 2002. Respectively, 5428, 4513 and 5645 respondents have filled in the questionnaires anonymously in the classroom according the methodology of international HBSC study. RESULTS: Boys were using alcohol on the regular basis (once a week or more often) more frequently than girls. Prevalence of regular alcohol consumption has increased from 9.4 to 13.6% in boys and from 4.2% to 6.5% in girls (p<0.05). Beer was the most popular alcohol drink among the students and was consumed respectively by 8.9%, 10.2% and 12.7% boys, and by 3.4%, 4.6%, 5.4% girls at least once per week. Prevalence of respondents who have reported the drunkenness (two or more times) has increased in boys from 13.3% to 30.1%, and in girls from 27.1% to 19.6% during period 1994-2002 (p<0.05). CONCLUSIONS: Trend of increase of alcohol consumption among school-aged children was observed in Lithuania during 1994-2002. Boys tend to be more frequent regular users of alcohol than girls. Alcohol consumption was significantly increasing with age. Alcohol use behaviors of Lithuanian school-aged children tend to be closer to countries of the Western Europe and the Nordic countries during the period of observation. PMID- 15547315 TI - [Diagnosis of acute respiratory failure and nosocomial pneumonia]. AB - The aim of this study was to determine diagnosis and factors influencing acute respiratory failure and nosocomial pneumonia according to literature and clinical findings in critically ill patients. The term "respiratory failure" implies the inability to maintain either normal delivery of oxygen to tissues or normal removal of carbon dioxide from the tissues. There are many patients suffering from acute respiratory failure caused by nosocomial pneumonia, septic syndrome, aspiration, interstitial or alveolar lung edema, thromboembolism of a. pulmonalis, polytrauma and contusion of the lungs, acute respiratory distress syndrome, acute lung injury, status asthmaticus, rather massive transfusions of blood products, and lipid embolism in the intensive care unit. There are actually three processes involved: transfer of oxygen across the alveolus, transport to the tissues (by cardiac output), and removal of carbon dioxide from the blood into the alveolus with subsequent exhalation into the environment. Failure of any step in this process can lead to respiratory failure. Long-term hypoxia causes ischemic changes and dysfunction of brain, heart, kidney, lungs and can worsen the outcome of disease or can cause higher mortality. PMID- 15547316 TI - Functional foods, herbs and nutraceuticals: towards biochemical mechanisms of healthy aging. AB - Aging is associated with mitochondrial dysfunctions, which trigger membrane leakage, release of reactive species from oxygen and nitrogen and subsequent induction of peroxidative reactions that result in biomolecules' damaging and releasing of metals with amplification of free radicals discharge. Free radicals induce neuronal cell death increasing tissue loss, which could be associated with memory detriment. These pathological events are involved in cardiovascular, neurodegenerative and carcinogenic processes. Dietary bioactive compounds from different functional foods, herbs and nutraceuticals (ginseng, ginkgo, nuts, grains, tomato, soy phytoestrogens, curcumin, melatonin, polyphenols, antioxidant vitamins, carnitine, carnosine, ubiquinone, etc.) can ameliorate or even prevent diseases. Protection from chronic diseases of aging involves antioxidant activities, mitochondrial stabilizing functions, metal chelating activities, inhibition of apoptosis of vital cells, and induction of cancer cell apoptosis. Functional foods and nutraceuticals constitute a great promise to improve health and prevent aging-related chronic diseases. PMID- 15547317 TI - Single nucleotide polymorphisms: aging and diseases. AB - Differences of more than 3 million nucleotides can bee seen comparing the genomes of two individuals as a result of single nucleotide polymorphism (SNP). More and more SNPs can be identified and it seems that these alterations are behind of several biological phenomena. Personal differences in these nucleotides result for example in elevated disease susceptibilities, that is, certain nucleotides are more frequent in patients suffering from different diseases comparing to the healthy population. SNPs may cause substantial alterations in the cells, e.g. the enzyme activity of the respective gene changes, but in other cases the effects of the SNPs are not so pronounced. Later results indicate that SNPs can be rendered to individuals living a longer life than the average. Perhaps these results will not directly lead to the lengthening of the maximal life span; however, genes that play an important role in the aging process could be identified. In this respect SNPs are important factors in determining the information level of the cells of individuals which determines the maximal life span (I. Semsei On the nature of aging. Mech. Ageing Dev . 2000; 117: 93-108), in turn SNP is one of the factors that determine the aging process. Since there are certain age-related diseases, the discovery and the description of the SNPs as a function of age and diseases may result in a better understanding of the common roots of aging and those diseases. PMID- 15547318 TI - The histone acetyltransferase GCN5 modulates the retrograde response and genome stability determining yeast longevity. AB - Transcriptional silencing decreases at both subtelomeric and silent mating-type loci and increases at the ribosomal DNA locus during the replicative life span of the yeast Saccharomyces cerevisiae . Evidence exists that epigenetic changes in the regulatory state of chromatin may be a causal factor in determining yeast longevity and that histone deacetylases play a role. The significance of histone acetylation has been examined here in more detail. Deletion of the histone acetyltransferase gene GCN5 suppressed the extension of replicative life span afforded by the induction of the retrograde response, which signals mitochondrial dysfunction and leads to changes in nuclear gene expression. It was difficult to ascribe this effect to changes in transcriptional silencing in any of the three known types of heterochromatin. However, a promoter related effect was uncovered by the participation of GCN5 in the induction of the retrograde response. Gcn5p and the retrograde signal transducer Rtg2p are components of the histone acetyltransferase coactivator complex SLIK. Rtg2p blocks the production of extrachromosomal ribosomal DNA circles when it is not engaged in transmission of the retrograde signal. Deletion of GCN5 , which disrupts the integrity of SLIK, suppressed circle accumulation. The results indicate that Gcn5p and SLIK impact the interplay between the retrograde response signal and Rtg2p with consequences for the induction of the response and circle production. Rtg2p and Gcn5p in the SLIK complex link metabolism to stress responses, chromatin-dependent gene regulation, and genome stability in yeast aging. PMID- 15547319 TI - D-galactose-caused life shortening in Drosophila melanogaster and Musca domestica is associated with oxidative stress. AB - D-galactose causes aging acceleration in different animal models but the mechanism is unclear. In the present study, we investigated the effects of D galactose on lifespan and oxidative stress biomarkers in the fruit fly (Drosophila melanogaster) and housefly (Musca domestica). D-galactose was added to drinking water (20 mg/ml) for housefly and to culture medium (6.5%) for fruit fly from 24 h after emergence. Oxidative stress was estimated by measuring the activity of Cu-Zn-superoxide dismutase (SOD) and the levels of lipid peroxidation products, namely malondialdehyde (MDA) and lipofuscin in housefly brain (male) and in fruit fly (male and female). D-galactose caused a significant decrease in mean lifespan (by 12.6% of male and 15.9% of female) and maximum lifespan (by 12.9% of male and 17.1% of female) in fruit fly, and also a significant decrease in mean lifespan (by 27.1% of male, 19.8% of female) and maximum lifespan (by 27.1% of male, 21.9% of female) in housefly. MDA and lipofuscin increased with age in fruit fly and in housefly brains while change of the SOD activity showed a biphasic shape with age. D-galactose caused a significant increase in MDA and lipofuscin and decrease in SOD activity in the age-matched fruit flies and houseflies. These data indicate that D-galactose shortens the lifespan of the two different fly species and that the life shortening effect is associated with an increase in oxidative stress. PMID- 15547320 TI - Cross-life stage and cross-generational effects of gamma irradiations at the egg stage on Drosophila melanogaster life histories. AB - The long-term effects of X-irradiation with 0.25, 0.5, 0.75 and 1 Gy of 1 h eggs on the fitness-related life history traits in adult Drosophila melanogaster fruit flies and their offspring were investigated. Following irradiation with 0.25, 0.5 and 0.75 Gy, both F0 and F1 flies have decreased adult body weight and increased locomotor (photo- and geotactic) activity, whereas metabolic rate measured as the rate of CO2 production was unchanged or even increased, and female fecundity was slightly reduced compared to appropriate controls. In some cases, irradiation resulted in hormetic effects increased resistance to both starvation and heat shock stresses as well as life extension. An explanation of the beneficial long lasting effects induced by early irradiation is offered, which suggests that these effects are due to cross-life stage and cross-generational adaptive phenotypic plasticity. PMID- 15547321 TI - Pineal graft in old rats improves erythrocyte resistance to peroxyl radical induced hemolysis. AB - Pineal graft from young to old rats was performed and red blood cell hemolysis, induced by the water-soluble radical initiator 2,2'-azobis (2-amidino-propane) dihydrochloride (AAPH), was evaluated 6 months after graft. Pineal graft modified the hemolysis curve kinetic profile in grafted rats versus age-matched controls, the 50% hemolysis time as well as the lag time were longer, whereas the maximal amount of hemolysis was lower, and it occurred over a longer period of time. Thiobarbituric acid reactive substances production was lower in pineal-grafted rats than in controls and the age-related decrease of erythrocyte membrane fluidity was prevented by pineal graft. The present findings support an important physiological role of pineal gland in preventing age-related alterations of erythrocyte membranes and suggest a possible antioxidant action of melatonin. PMID- 15547322 TI - Age-related changes of insulin receptors, plasma insulin and glucose level. AB - The effects of aging on hepatic and erythrocyte insulin receptors have been investigated in 6, 12, 18 and 21-months-old compare to 3-months-old rats. Plasma insulin was elevated in 6, 12 and 18-months-old rats. Specific binding of insulin in liver was increased at the age of 8 months and accompanied with increase in concentration of low affinity binding sites, while specific binding to erythrocytes as well as concentration of both classes of binding sites was increased in 6-months-old rats. The protein and mRNA content of hepatic receptor were decreased only in the oldest animals. Plasma glucose was elevated starting from 12-months-old rats, while, after decrease in 6-months-old animals, citrulline was raised in the oldest group. The results demonstrating that specific binding of insulin in liver and erythrocytes and the concentration of binding sites in both tissues were not decreased during aging, as well as the absence of changes in affinity of insulin binding sites do not point out to occurrence of insulin resistance. However, the increase in insulinemia in the middle of lifespan, elevated plasma glucose and citrulline as well as decrease of hepatic receptor protein and mRNA content in the oldest animals indicate some age related changes in insulin signaling. PMID- 15547323 TI - Age-dependent alterations in mitochondrial enzymes in cortex, striatum and hippocampus of rat brain -- potential role of L-Carnitine. AB - Mitochondria link the energy -- releasing activities of electron transport and proton pumping with the energy conserving process of oxidative phosphorylation to form ATP. A declined mitochondrial performance has been generally observed during aging. In the present investigation, the activities of tricarboxylic acid cycle enzymes such as isocitrate, alpha-ketoglutarate, succinate and malate dehydrogenases and electron transport complexes I-IV were measured in mitochondria isolated from brain regions like cortex, striatum and hippocampus of young and aged rats before and after L-Carnitine supplementation. All the three brain regions of aged rats showed decreased activities of isocitrate, alpha ketoglutarate and succinate dehydrogenases, complexes I and IV when compared to control young rats. Striatum seems to be the most susceptible region when compared to hippocampus and cortex. L-Carnitine supplementation to aged rats reversed the activities of these enzymes to near normal whereas treatment to young rats did not show any significant alterations. These results confirm that L Carnitine can alleviate the age-associated decline in the metabolic efficiency of mitochondria in all three brain regions under investigation. PMID- 15547324 TI - Meiotic recombination: an affair of two recombinases. AB - In E. coli, homologous recombination is catalyzed by the RecA recombinase. Two RecA-like factors, Rad51 and Dmc1, are found in eukaryotes. Whereas Rad51 is needed for homologous recombination reactions in both mitotic and meiotic cells, the role of Dmc1 is restricted to meiosis. Recent work has shown that, like RecA and Rad51, Dmc1 mediates the homologous DNA pairing strand exchange reaction via a filamentous intermediate assembled on single-stranded DNA. Emerging evidence suggests that the tumor suppressor BRCA2 functions in the assembly of nucleoprotein filaments of Rad51 and Dmc1. The manner in which Rad51 and Dmc1 functionally cooperate in meiotic recombination remains to be determined. PMID- 15547370 TI - Health care delivery to the patients and the evolving systems of disruptions. PMID- 15547371 TI - From past to present and future is today: from inert to multifunctional biomaterials. PMID- 15547372 TI - The molders of this plastic surgeon and his quest for symmetry. PMID- 15547373 TI - Fibrous dysplasia of the face: utility of three-dimensional modeling and ex situ malar recontouring. AB - A 20-year-old male with fibrous dysplasia involving the right fronto-orbital and malar regions showed no significant progression of disease by serial computed tomography (CT) scans over 6 years. Two prior attempts to recontour the right maxilla and zygoma in situ and to increase the right orbital volume had been unsatisfactory. To solve the problem of persistent right-sided proptosis and facial asymmetry, a three-dimensional model of the midface and orbits was made on the basis of a pre-operative CT scan. Removable components transformed the model of the affected right side of the midface into a mirror image of the unaffected side, giving a precise indication of where and how much bone needed to be removed. Surgical correction was performed using a right malar osteotomy in which the zygoma was mobilized in continuity with the lateral and inferior orbital rims. This approach gave direct access to the orbit, through which the lateral and medial orbital walls were re-contoured to increase intraorbital volume. The freely mobilized zygoma was then shaped to match the prefabricated model. The zygoma was repositioned, and the affected maxilla was recontoured to blend with the remodeled zygoma. Follow-up CT scan and physical examinations postoperatively demonstrate excellent facial symmetry and correction of right-sided proptosis. When there is no documented progression of fibrous dysplasia in the face over several years, three-dimensional modeling as a guide to ex situ malar recontouring can improve the accuracy of facial reconstruction. This approach also provides direct access to the orbit for the correction of bony orbital volume. PMID- 15547374 TI - Role of guided bone regeneration principle in preventing fibrous healing in distraction osteogenesis at high speed: experimental study in rabbit mandibles. AB - The formation of fibrous tissues at the distraction gap may result from the accumulation of rapidly migrating fibroblasts at the site of an osteotomy, especially when distraction is rapid. Addition of osteopromotive membranes could theoretically prevent fibroblasts from entering the distraction gap, allowing the osteotomy site to be filled with only osteogenic cells. This study is an attempt to achieve a rapid successful distraction without fibrosis through the use of collagen membranes. Sixteen skeletally mature New Zealand white rabbits were used in this study. They were divided into two groups. One rabbit from each group was excluded from the study because of dislodgement of the distractors. In one group (n = 7), distraction was done as usual. In the other group (n = 7), a collagen membrane surrounded the osteotomy site to be distracted. After a 7-day latency period, distraction started at a rate 2 mm once per day for 5 days. The distractor was left in place for 4 weeks to allow consolidation. Results showed osteogenesis in both groups. Whereas addition of the membrane to distraction increased the quantity of bone formed, absence of the membrane allowed early mineralization (better quality of bone regarding the density). Neither of the two groups showed significant fibrosis or cartilage formation. The endosteum served as a source of blood supply when the periosteum was excluded. The periosteum served as a membrane for guided bone regeneration. Membranes for guided bone regeneration can be used with distraction when the periosteum is lost from trauma or is broken from fast distraction. PMID- 15547375 TI - Immunolocalization of androgen receptor in the developing craniofacial skeleton. AB - Male predominance in metopic and sagittal craniosynostosis and in nonsynostotic plagiocephaly suggests a role for circulating androgens in early craniofacial development. Androgens have been documented to play an important role in postnatal skeletal growth, and the androgen receptor has been recently demonstrated in human and rat osteoblast-like cell lines and in human long bones. The purpose of this study was to describe the expression of androgen receptor in the fetal craniofacial skeleton. The heads of E18 fetal CD-1 male and female mice were fixed in 10% formalin, decalcified, and embedded in paraffin. Four- to 6-mum coronal and sagittal sections were stained with a monoclonal antibody specific to androgen receptor, which was detected by an avidinbiotin conjugate and peroxidase system. The sections were then examined for androgen receptor expression patterns. Strong androgen receptor immunoreactivity was observed in the dura mater of developing fetuses. Androgen receptor expression was also noted in cells lining the osteogenic fronts and in calvarial osteoblasts. Similar androgen receptor expression patterns were found in male and female mice. Androgen receptor is abundantly expressed in fetal dura mater and calvarial bone. This study confirms the presence of androgen receptor in the murine fetal craniofacial skeleton, suggesting a potential role for the anabolic effects of androgens in the developing craniofacial skeleton. PMID- 15547377 TI - Autogenous orbital reconstruction in a child with congenital abnormalities of the orbital roof and vertical orbital dystopia. AB - Congenital anomalies of the orbital roof are rare occurrences. The case of a 2 year-old child with vertical orbital dystopia and abnormalities of the right bony orbit is presented. The patient underwent right orbital reconstruction to restore facial symmetry. A coronal approach with a frontal craniotomy was used for intracranial exposure. The abnormal angulation of the roof was corrected, and the defect was reconstructed with a split-calvarial bone graft harvested from the parietal region. The bone graft was secured with resorbable plate fixation. To preserve vision, reconstruction of this type must be done at an early age, preferably before the age of 4 years. In this patient, there is good facial symmetry and normal globe positioning 5 years after surgery. PMID- 15547378 TI - Swallowing disorders in Pierre Robin sequence: its correction by distraction. AB - Pierre Robin sequence is characterized by micro-gnathia, glossoptosis, feeding difficulties, and upper respiratory obstruction, which are frequently complicated by bronchial aspiration and pulmonary infection. Gastroesophageal reflux is also common in these patients. To assess the results of mandibular distraction, a study was performed in 18 patients to detect swallowing disorders associated with apnea episodes and gastroesophageal reflux. Polysomnography, barium pharyngoscopy, determination of blood gases, and esophageal pH measurements were undertaken before and 4 months after distraction osteogenesis. Bilateral corticotomies, followed by distraction with external devices, were performed, achieving 7 to 19 mm of elongation (mean = 12 mm). Gastroesophageal reflux was found in 83% of cases associated with apnea episodes, but it disappeared after distraction osteogenesis. Mean preoperative oxygen saturation was 72%, and it was 93% afterward. The preoperative apnea index was 18.3, and the preoperative 8.5 hypopnea index was 8.5; both disappeared. Pharyngeal transit time became less than 1 second after treatment. Abnormal tongue movements and barium stasis in the pharyngeal recess and in the trachea were eliminated in all the patients. PMID- 15547379 TI - Blepharoptosis caused from compression of levator muscle by fractured orbital roof fragment. AB - The authors report two cases of blepharoptosis caused by compression of the levator muscle by fractured orbital roof fragment; both were improved by surgical treatments. In one case, because neurosurgeons were concerned with the contusional hemorrhage of the right frontal lobe during the early admission period, they missed the orbital roof fracture. The patient underwent surgery on the 18th post-trauma day. In the other case, the early diagnosis was made, and the surgical treatment were performed on the 10th post-trauma day. The authors think if the blepharoptosis is not improved in several days and the fractured fragment is suspected to compress the levator in computed tomography scan, surgical decompression is necessary and should be performed as soon as possible. PMID- 15547380 TI - "Cat's cradle" midfacial fixation in distraction osteogenesis after Le Fort III osteotomy. AB - Distraction osteogenesis has provided a powerful technique for the treatment of severe midfacial hypoplasia. Skeletal fixation is a critical component of successful distraction osteogenesis. This is a report on the development of a new approach to bone fixation for distraction osteogenesis of the midface, based on circumferential suspension of the entire midfacial skeleton using surgical wires placed through the soft palate. This technique offers the advantages of (1) complete control of the attitude of the midface in terms of a precise dispersion of forces that can determine the desired pitch, yaw, and roll of this skeletal complex during the active phase of distraction, (2) being compatible with young children in the deciduous dentition stage because it obviates the need for rigid fixation devices on the maxillary lateral walls in the proximity of numerous dental follicles of the unerupted adult dentition, (3) actually serving as the instrument for posterior maxillary "lift" in cases of skeletal open bite malocclusions (frequently seen in the craniosynostoses), thereby allowing the clinician to determine the final occlusal and palatal planes and simultaneously close the open bite. An illustrative case of a 5(1/2)-year-old girl with Apert syndrome is presented wherein her ventilatory obstruction and ocular proptosis were significantly improved. PMID- 15547381 TI - Hybrid of distraction osteogenesis unilateral frontal distraction and supraorbital reshaping in correction of unilateral coronal synostosis. AB - Unicoronal synostotic plagiocephaly is routinely treated by intracranial wide frontal and bilateral supraorbital reshaping. Recent advancement of distraction osteogenesis in craniofacial surgery has extended to patients with craniosynostosis. Although a controversy remains between conventional osteotomy and reshaping and application of the distraction technique in surgical treatment of craniosynostosis, there have been several positive clinical reports on distraction techniques for nonsyndromic and syndromic craniosynostosis. Unicoronal distraction applied successfully to a case of frontal plagiocephaly has been described. The authors report a procedure: hybrid of unilateral frontal distraction and supraorbital reshaping on the affected side for frontal synostotic plagiocephaly. This procedure was conducted on four typical unicoronal synostotic plagiocephaly cases (patient age range, 9-14 months; all patients female) with successful results. No particular complications were encountered in any of the four cases with a follow-up period that ranged from 18 to 53 months. PMID- 15547382 TI - Enhanced frontalis sling with double-fixed, solvent-dehydrated cadaveric fascia lata allograft in the management of eye ptosis. AB - The article describes an enhanced frontalis sling procedure in the management of blepharoptosis with poor or no levator function. This modified method were used in the care of three patients and based on the principal of two-point fixation of the graft material that was prepared from solvent-dehydrated cadaveric fascia lata onto the frontalis muscle. After more than 1 year of follow-up, no recurrence was noted. With no extra incision over the skin, this surgical approach could be useful in the correction of difficult ptosis cases. PMID- 15547384 TI - Frontal sinus obliteration with beta-tricalcium phosphate . AB - Autogenous materials are considered the gold standard when indicated for frontal sinus obliteration; however, alloplastic alternatives obviate the need for a second surgical site, eliminate donor site morbidity, and reduce operating room time. A variety of different alloplastic materials have been used for this purpose, but no report to date has described the use of pure beta -tricalcium phosphate synthetic bone. This case report describes the technique and successful use of this product for frontal sinus obliteration in a severely comminuted anterior table and nasofrontal duct injury. The product warrants consideration as a viable alternative to standard autogenous and alloplastic substrates for frontal sinus obliteration. PMID- 15547385 TI - Intra- and perioperative complications of the LeFort I osteotomy: a prospective evaluation of 1000 patients. AB - The LeFort I osteotomy has become a routine procedure in elective orthognathic surgery. The authors report the occurrence of intra- or perioperative complications in a series of 1000 consecutive LeFort I osteotomies performed within a 20-year period. In total, 64 (6.4%) patients experienced complications. Anatomical complications affected 26 (2.6%), patients, including 16 (1.6%) with a deviation of the nasal septum and 10 (1.0%) with non-union of the osteotomy gap. Extensive bleeding that required blood transfusion occurred in 11 (1.1%) patients exclusively after bimaxillary corrections; in 1 patient a ligation of the external carotid artery became necessary. Significant infections such as abscesses or maxillary sinusitis occurred in 11 (1.1%) patients. No patient experienced an osteomyelitis. Ischemic complications affected 10 (1.0%) patients, including 2 (0.2%) who experienced an aseptic necrosis of the alveolar process and 8 (0.8%) who, under critical revision, were affected by retractions of the gingiva. Five (0.5%) patients experienced an insufficient fixation of the osteosynthesis material. The risk and the extent of complications was enhanced in patients with anatomical irregularities (eg, in patients with craniofacial dysplasias, orofacial clefts, or vascular anomalies). The risk of ischemic complications was enhanced in extensive dislocations or transversal segmentation of the maxilla. The authors conclude that patients with major anatomical irregularities should be informed about an enhanced risk of Le-Fort I osteotomies. Preoperative planning avoiding transversal segmentation or extensive dislocations of the maxilla should reduce the occurrence of complications. For healthy individuals, the risk of complications with the LeFort I osteotomy is considered low. PMID- 15547387 TI - Intraosseous hemangioma of the maxillofacial district: clinical analysis and surgical treatment in 10 consecutive patients. AB - The aims of this study are to illustrate functional and esthetic results obtained with different surgical strategies and to report a review of the relevant literature. There were 6 female patients and 4 male patients included in this study, with an average age of 35.7 years. Zygomatic bone was affected in six cases, the mandible in two cases, the medial orbital wall in one case, and the upper jaw in one case. In all 10 patients, surgery consisted of a wide excision of the intraosseous hemangioma with margins of 3 mm at least to ensure complete removal. Immediate reconstruction was carried out in 5 of the 10 patients. An analysis indicates that intraosseous hemangiomas of the maxillofacial area are rare; diagnosis can be difficult and is mainly based on computed tomography scans. Surgical excision, with previous angiography and embolization in cases of intraosseous hemangioma with a larger dimension or abnormal blood supply, is the treatment of choice. PMID- 15547388 TI - Classification system and treatment of zygomatic arch fractures in the clinical setting. AB - A new classification system and algorithm of zygomatic arch fractures is described that provides the surgeon with a useful starting point from which to organize a valid treatment plan and management of zygomatic arch fractures. Honig Merten (HM) class I is defined as an isolated tripod fracture, HM class II as an isolated stick fracture of the arch, and HM class III is a combined fracture of the malar bone and the zygomatic arch. Although reduction of the class I and II is usually closed, open reduction is mandatory in class III zygomatic arch fractures. PMID- 15547389 TI - Monobloc distraction osteogenesis in pediatric patients with severe syndromal craniosynostosis. AB - The management of the hypoplastic midface in syndromic craniosynostosis remains a great challenge. Frequently, patients have to be operated on numerous times to achieve a satisfactory end result, partially because of the limited skeletal advancement possible when using traditional surgical techniques. During the last decade, however, methods for gradual midfacial distraction have been presented, whereby greater advancements can be obtained. We present four children aged 17 months to 15 years with severe syndromal craniosynostosis in need of midface advancements because of severe respiratory obstruction or severe exophthalmos. These patients were complex cases with several previous craniofacial surgeries (mean of three times, range of two to six times) that yielded insufficient skeletal advancements. They were operated on with gradual monobloc advancements using the Modular Internal Distraction System. The mean length of operations was 370 minutes (range: 240-455 minutes), and the mean amount of perioperative blood transfusion needed was 1,300 ml (range: 280-2,700 ml) or 66.9 ml/kg (range: 31.1 94.9 ml/kg). The patient with the greatest number of previous operations also had the longest operation time as well as the most blood loss. The average midface advancement obtained was 25 mm (range: 20-30 mm), resulting in cessation or a significant decrease of preoperative respiratory problems, reduced exophthalmos, and improved facial profile. Apart from a local infection in one patient with a connective tissue disorder and several previous wound infections, no major postoperative complications were recorded. Distraction osteogenesis has become a versatile and safe technique that allows for large advancements of the midface. PMID- 15547391 TI - Free fibula flap mandible reconstruction in benign mandibular lesions. AB - The mandible may be affected by a variety of pathological conditions such as ameloblastomas, odontogenic keratocysts, central giant cell granulomas, fibro osseous lesions, and osteomas. They generally are benign, and conservative treatment can be enough in most of the cases. However, they can be clinically aggressive, and especially ameloblastomas, odontogenic keratocysts, and fibro osseous lesions are prone to recur. The method of treatment remains a matter of controversy in these cases. The aim of this analysis was to study 100 patients who had been treated for benign mandibular lesions, and to emphasize the importance of free fibula flap in the treatment of such lesions. PMID- 15547392 TI - Phenol intoxication in a child. AB - Phenol preparations are used in dermatology and plastic surgery for the treatment of acne and during chemical face peeling. At this institution, phenol peeling is used in addition to mechanical dermabrasion for the elimination of subclinical premalignant lesions of patients having xeroderma pigmentosum. As the phenol peel is performed, most surgeons concentrate on skin results, ignoring systemic complications. Local histological changes and systemic toxicity have been seen during applications. Cardiac arrhythmias and even sudden death have been reported. The high incidence of cardiac arrhythmias after topical application of phenol preparations is demonstrated. The case of an 11-year-old boy with a diagnosis of xeroderma pigmentosum who underwent mechanical dermabrasion and chemical peeling with phenol and then developed severe cardiac arrhythmias is reported. A serious systemic toxic effect on cardiac rhythm from cutaneously applied phenol occurred in this case. PMID- 15547393 TI - Intracranial volume measurement of metopic craniosynostosis. AB - The authors report 32 patients with nonsyndromic isolated metopic synostosis who have undergone evaluation of their intracranial volumes. Twenty-five were male and seven were female. The measured intracranial volumes were compared with normal age-corrected values established in the authors' unit, and any differences were noted. The authors found that, although there was a range of intracranial volumes, in the male patients, intracranial volumes were significantly smaller than those found in the normal population (P < 0.05). However, this result was not found in the smaller female sample. These results contrast with those of smaller earlier studies, but the authors conclude that intracranial volumes are smaller than average for age-corrected normal values; this finding highlights the need for volume expansion in conjunction with cranial reshaping. PMID- 15547395 TI - Visual loss in syndromic craniosynostosis with papilledema but without other symptoms of intracranial hypertension. AB - Four children with syndromic craniosynostosis (Crouzon's syndrome and Apert syndrome) developed irreversible visual loss. Apart from papilledema, there were no other clinical symptoms of intracranial hypertension. Ventriculomegaly or hydrocephalus was present in all cases. Two children were known to have obstructive sleep apnea syndrome. These cases are reported to inform specialists involved in follow-up of children with syndromic craniosynostosis that visual loss can develop suddenly without other symptoms of intracranial hypertension. To prevent visual loss, papilledema should be detected at an early stage and intervention should be instituted promptly. Therefore, periodic funduscopy should be performed in children at risk, such as children with syndromic craniosynostosis and additional hydrocephalus or obstructive sleep apnea syndrome. PMID- 15547397 TI - The "sardine can technique": a method for the removal of internal fixation hardware. PMID- 15547398 TI - Pleomorphic adenoma of the palate. AB - Pleomorphic adenoma, is the most common tumor (50%) of the major and minor salivary glands. Seventy percent of the tumors of the minor salivary glands are pleomorphic adenomas, and the most common intraoral site is the palate, followed by the upper lip and buccal mucosa. Pleomorphic adenoma appears as a painless firm mass and, in most cases, does not cause ulceration of the overlying mucosa. Generally it is mobile, except when it occurs in the hard palate. Intraoral mixed tumors, especially those noted within the palate, lack a well-defined capsule. Lesions of the palate frequently involve periosteum or bone. Approximately 25% of benign mixed tumors undergo malignant transformation. Treatment for the pleomorphic adenoma is radical surgery. Inadequate resection leads to local recurrence. The authors report a palate pleomorphic adenoma in a 67-year-old female patient. PMID- 15547399 TI - Aneurysmal bone cyst of the maxilla. AB - An aneurysmal bone cyst of the maxilla is a nonneoplastic, uncommon, solitary bone lesion recognized by distinct radiographic and histopathological characteristics. It is described as a localized and quickly expandable benign tumor, which can reach a considerable size. It is characterized by the replacement of bone by spongy fibro-osseous tissue and a locally destructive and multicystic lesion filled with blood. It is attributed to a circulatory disturbance leading to locally increased venous pressure but with an unclear etiology. It takes place mainly in the long bones and in patients less than 20 years old, with no sex predilection. Its treatment is by surgical excision. An aneurysmal bone cyst in the facial bone is rare. The present case describes the lesion arising in the maxilla of a 14-year-old girl. PMID- 15547400 TI - Extensive neglected psammomatoid ossifying fibroma with craniofacial deformity. AB - The psammomatoid ossifying fibroma is a rare and aggressive lesion that usually arises from the sinonasal region. The lesion is well circumscribed with the pathognomonic feature of concentric or laminated spherical ossicles that appear similar to psammoma bodies. The authors report a case of an extensive and neglected psammomatoid ossifying fibroma in a 10-year-old West African girl. She had a 5-year history of a slowly growing tumor, which at presentation was 15 cm in diameter and had resulted in left craniofacial deformity as well as left visual impairment. Due to the large tumor, the surgical resection plan necessitated a cranio-facial neurosurgical approach in combination with autogenous bone grafts for cranofacial reconstruction, and allogenous demineralized bone grafts for donor site coverage. In addition, preoperative embolization was utilized to reduce tumor vascularity. However, this embolization resulted in reduction of pericranial flap vascularity, leading to flap ischemia, infection and subsequent removal of necrotic autogenous bone graft. Ultimately, the objectives of large tumor resection and acceptable aesthetic outcome were met after controlling an episode of postoperative infection. PMID- 15547401 TI - Infantile fibrous dysplasia of the mandible. AB - Fibrous dysplasia of the craniofacial skeleton is an uncommon benign neoplasm. Its appearance in infancy (<1 year) is extremely rare. The authors report a case of mandibular fibrous dysplasia presenting in a patient 12 weeks of age as a painless, primarily left-sided, enlarging neoplasm of bone. Surgical contouring was successfully performed when the patient was 9 months of age. The authors anticipate recurrence, but at 6 months of follow-up no new growth has been observed. PMID- 15547402 TI - Correction of facial deformity using a Red III device in a patient with Antley Bixler syndrome. AB - Antley-Bixler syndrome was first described in 1975 and to date at least 44 cases have been reported. In addition to brachycephaly, this syndrome is associated with midface hypoplasia, bilateral radiohumeral synostosis, multiple joint contractures, long bone fractures, dysplastic ears, "pear shaped" nose, and occasionally urogenital or cardiac defects. The authors report a case in a 23 year-old man with severe midface deficiency treated using distraction osteogenesis. A modified Le Fort III osteotomy was performed, and nasal subunit and remaining Le Fort III segment was distracted separately. The classic RED II design was found to be insufficient, so the authors added an extra horizontal bar to the system and converted it to a RED III design. Midface advancement was 11 mm from the porion to the orbitale and 31 mm at point A. Nasal advancement at the nasal root level was 5 mm. There were no complications, and at 4 months after surgery, advancement was stable without any relapse. PMID- 15547403 TI - Postnatal onset of craniosynostosis in a case of Saethre-Chotzen syndrome. AB - Saethre-Chotzen syndrome is a craniosynostosis syndrome characterized by facial and limb abnormalities. It is caused by mutations in the TWIST gene on chromosome 7p21. To date, more than 80 different mutations in TWIST have been reported in the literature.Recently, large deletions of chromosome 7p, encompassing the TWIST locus, have been detected in patients with clinical features of Saethre-Chotzen syndrome. Strikingly, all these patients were severely mentally retarded, which is otherwise a rare finding in Saethre-Chotzen syndrome. The authors report a patient with a large TWIST/7p deletion but with normal development. Furthermore, craniosynostosis was not present at birth or at the age of 4 months. However, skull radiographs taken at the age of 14 months showed stenosis of both coronal sutures, as well as of part of the sagittal suture. Reports on postnatal onset of craniosynostosis have been made in Crouzon syndrome but, to the authors' knowledge, never in Saethre-Chotzen syndrome. PMID- 15547404 TI - Avascular necrosis of the mandibular condyle causing fibrous ankylosis of the temporomandibular joint in sickle cell anemia. AB - Avascular necrosis, which most commonly affects the femoral head, is a well recognized entity in orthopedic surgery. Maxillofacial surgeons have recently recognized that a similar process may also be at work in the temporomandibular joint because of the involvement of the mandibular condyle. Avascular necrosis of the mandibular condyle in association with sickle cell anemia is an infrequent finding. In this report, a patient with avascular necrosis of the mandibular condyle in the late stage causing fibrous ankylosis of the temporomandibular joint in sickle cell anemia is presented. PMID- 15547405 TI - Differential diagnosis between adenoid cystic carcinoma and pleomorphic adenoma of the minor salivary glands of palate. AB - Tumors arising from minor salivary glands of the palate may exhibit an overlap of clinical and biologic features that may produce diagnostic and therapeutic dilemmas. Surgical treatment can be very different, depending on the dimensions and malignant or benign nature of the tumors, and therefore should be planned on the basis of an accurate differential diagnosis. A retrospective analysis in 24 patients with pleomorphic adenoma and adenoid cystic carcinoma of minor salivary glands of the palate was performed to investigate the accuracy of fine needle aspiration cytology (FNAC) and biopsy with histology in the preoperative diagnosis. Preoperative diagnoses obtained with FNAC and biopsy were compared with findings of the definitive histopathologic examination performed on the resected mass. Correspondence between the preoperative diagnoses determined by FNACs and the definitive histopathologic results was observed in 22 of 24 cases, whereas a complete equivalence was found with regards to histology. In the analysis, FNAC was associated with 91.6% accuracy and an error rate of 8.4% in the diagnosis of pleomorphic adenoma and adenoid cystic carcinoma of the palate. From the results of the analysis, histologic examination is still the most accurate diagnostic tool in such tumors. FNAC can be considered in tumors of the head and neck regions that are difficult to reach by means of a common biopsy. PMID- 15547406 TI - Seizure as a cause of orocutaneous fistula. PMID- 15547407 TI - Melorheostosis involving the craniofacial skeleton. AB - Melorheostosis is a rare bone disorder, usually affecting the long bones and adjacent soft tissue. It was originally described by Leri and Joanny in 1922, after its classic x-ray features of flowing hyperostosis resembling dripping candle wax. There have been fewer than 10 reported cases of craniofacial involvement, and in most instances these have also involved the appendicular skeleton. The authors report a case of melorheostosis with isolated craniofacial involvement, describe the clinical course and radiologic and histologic features, and review the pertinent literature. PMID- 15547412 TI - A comparison of cartilage palisades and fascia in tympanoplasty after surgery for sinus or tensa retraction cholesteatoma in children. AB - OBJECTIVE: To compare cartilage palisades with fascia grafting in reconstruction of the eardrum after surgery for sinus or tensa retraction cholesteatoma in children, with respect to postoperative drum retraction and perforation, cholesteatoma recurrence, and hearing. MATERIAL: From March 1995 to October 2000, a total of 64 children, aged 5 to 15 years, underwent surgery for either sinus or tensa retraction cholesteatoma. The eardrum was reconstructed using cartilage palisades in 32 children and fascia or perichondrium in 32 children. Postoperatively, the patients were seen as out-patients and were recently reevaluated by otomicroscopy and audiometry. MAIN OUTCOME MEASURES: Postoperative drum retraction and perforation, cholesteatoma recurrence, and hearing (pure tone average, speech reception threshold, and air-bone gap). RESULTS: All patients in the palisade group and all but one patient in the fascia group attended the follow-up examination. In the palisade group, the mean follow-up period was 37 months (range, 3-65 mo) and in the fascia group 52 months (range, 17-75 mo). Two (6%) retractions and no perforations were found in the palisade group, versus 12 (36%) retractions and 4 perforations (12%) in the fascia group at follow-up (both significant differences). No cholesteatoma recurrence occurred. Late hearing results in sinus cholesteatomas were significantly better in the palisade group. CONCLUSIONS: The comparison of fascia and cartilage palisade grafting for drum reconstruction after tensa cholesteatoma surgery in children indicates that the palisade technique may be superior in respect to prevention of drum retraction and perforation. Further, in sinus cholesteatoma surgery, the long-term hearing results are better when grafting cartilage palisades. PMID- 15547413 TI - Eustachian tube patency and function in tympanoplasty with cartilage palisades or fascia after cholesteatoma surgery. AB - OBJECTIVE: To compare the Eustachian tube patency and function after tympanoplasty with either cartilage palisades or fascia grafting after one-stage surgery in children with tensa cholesteatoma. STUDY DESIGN: In children operated for tensa cholesteatoma, cartilage palisade tympanoplasty was performed in 32 ears and fascia tympanoplasty in 29 ears. The patency of the Eustachian tube was evaluated by the Valsalva maneuver before the operation, 1 to 3 months after the operation, and at a follow-up examination 46(1/2) months later (median). Eustachian tube function was evaluated by the nine-step inflation/deflation tympanometric test and the Toynbee test at the follow-up examination. Otomicroscopy and hearing evaluation were performed before and after surgery as well as at the follow-up. MAIN OUTCOME MEASURES: Eustachian tube patency and function. Hearing, postoperative eardrum perforation/retraction, and cholesteatoma recurrence. RESULTS: The Valsalva maneuver was positive in 30% of the ears before the operation, in 65% primarily after the operation, and in 78% at the follow-up examination. A poor tubal function was found in 57% at follow up. Overall, the late functional hearing results were better in ears with a positive Valsalva maneuver. There were no differences in tubal patency or function in relation to graft material, cholesteatoma, and tympanoplasty type. In ears with a poor tubal function, the hearing results were significantly better in the palisade group (63% success), compared with the fascia group (17% success). All of the four perforations, most of the retractions, and a single moist eardrum were found in the fascia group at the reevaluation. We found no correlation between the condition of the eardrum and the Eustachian tube function at the last evaluation. However, in ears with a poor tubal function, a nonretracted, nonperforated drum was found with higher frequency in the palisade group. Decision matrix analysis showed that the last postoperative Valsalva maneuver was the best predictor of the drum condition at the reevaluation. CONCLUSIONS: The Eustachian tube patency frequently improves after tympanoplasty after cholesteatoma surgery in children, regardless of graft material. The patency and function of the Eustachian tube seem to be without relation to graft material, cholesteatoma, or tympanoplasty type. Cartilage palisade tympanoplasty may be a better reconstruction technique after cholesteatoma surgery, especially in ears with a poor tubal function. PMID- 15547414 TI - Postauricular periosteal-pericranial flap for mastoid obliteration and canal wall down tympanomastoidectomy. AB - OBJECTIVE: To describe an effective technique for mastoid cavity obliteration in canal wall down tympanomastoidectomy for chronic otitis media and review its efficacy in producing a dry, low-maintenance, small mastoid cavity. DESIGN: : Retrospective clinical study of a consecutive series of procedures from 1995 to 2000. SETTING: Tertiary referral center and institutional academic practice in otology and neurotology. PATIENTS: Sixty consecutive procedures for active chronic otitis media with a minimum follow-up of 12 months (mean, 31 mo; range, 12-80 mo). INTERVENTION: All patients had canal wall down mastoidectomy with simultaneous tympanoplasty including split-thickness skin grafting. An inferiorly pedicled, periosteal-pericranial flap was used in conjunction with autologous bone pate to obliterate the mastoid cavity. The additional length provided by the pericranial extension of the flap permitted it to reach superior to the lateral canal and into the sinodural angle, with improved coverage of bone pate and better reduction of cavity size. OUTCOME MEASURES: The primary outcome measure was control of suppuration and creation of a dry, low-maintenance mastoid cavity, which was assessed using a previously developed semiquantitative scale. This scale includes a temporal dimension to assess control of infection. Secondary outcome measures included postoperative complications (i.e., hematoma, infection, flap necrosis, and meatal stenosis) and incidence of recurrent or residual cholesteatoma. RESULTS: Forty-nine ears (82%) maintained a small, dry, healthy mastoid cavity. Five ears (8%) had intermittent otorrhea easily controlled by topical treatment. Six ears (10%) had suboptimal control of otorrhea, of which four had meatal stenosis. There were no residual or recurrent cholesteatomas. Outcomes remained stable over progressively longer follow-up, up to 80 months. CONCLUSION: Obliteration of a canal wall down mastoid cavity by a postauricular periosteal-pericranial flap with autologous bone pate is a reliable and effective technique that results in a dry, trouble-free mastoid cavity in 90% of patients with active chronic otitis media. PMID- 15547415 TI - Detection of postoperative relapsing/residual cholesteatomas with diffusion weighted echo-planar magnetic resonance imaging. AB - PURPOSE: To assess the capability of echo-planar diffusion-weighted magnetic resonance imaging (MRI) (EPI-DWI) in diagnosing relapsing/residual cholesteatomas after canal wall-up mastoidectomy. MATERIALS AND METHODS: In a blinded study design, we investigated with MRI, including standard spin-echo sequences, 18 patients evaluated with clinical examination and computed tomography (CT) suspected for relapsing/residual cholesteatoma 7 to 19 months after a canal wall up mastoidectomy. Images were evaluated by two radiologists blinded to patients' identities, CT findings, and clinical data set, who decided in a consensus agreement whether there was a pathologic signal increase in the petrous bone in a single-shot EPI-DWI sequence. All the patients underwent a second tympanoplasty or revision surgery of the mastoidectomy cavity within 15 days after magnetic resonance investigation.Sensitivity, specificity, and predictive values were evaluated separately for standard sequences and EPI-DWI. RESULTS: In EPI-DWI, five of six patients with cholesteatoma showed a bright signal, whereas those patients with a noncholesteatomatous tissue showed no anomalies. The only misdiagnosed cholesteatoma was a pearl 2 mm in diameter. Sensitivity, specificity, and positive predictive values, and negative predictive values of EPI-DWI in diagnosing relapsing/residual cholesteatomas were 86, 100, 100, and 92%, respectively. CONCLUSION: EPI-DWI may be a useful tool in differentiating between cholesteatomatous and noncholesteatomatous tissues after closed cavity mastoidectomy. Further investigations are, however, required to establish the practical utility of EPI-DWI on larger series as a screening modality in the follow-up after closed cavity mastoidectomies. PMID- 15547416 TI - The Yung percutaneous mastoid vent: results of a multicenter trial. AB - OBJECTIVE: The primary goal of the study was to find out whether the Yung vent would remain patent long-term. The secondary goal was to find out whether mastoid ventilation could overcome permanent ventilation disorder within the middle ear. STUDY DESIGN: The study was an open prospective investigation. SETTING: The study was a multicenter study involving three tertiary referral centers. PATIENTS: Twenty-three subjects older than 12 years were included. The inclusion criteria were complete atelectasis, failed tympanoplasty because of persistent eustachian tube dysfunction, and persistent otitis media with effusion in spite of repeated ventilation tube insertion. INTERVENTION: Tympanoplasty with insertion of the Yung percutaneous mastoid vent. MAIN OUTCOME MEASURES: Patency of the vent, adverse effects, patient acceptance, hearing results, and appearance of the tympanic membrane were assessed every 3 months up to 18 months postoperatively. RESULTS: : At 18 months, 20 of 23 vents were still patent. Overall, there had been no unacceptable adverse effect on any patient. Eighteen of 23 ears had improved hearing. The tympanic membrane had returned from a collapsed state to near normal in 13 of 17 completely atelectatic ears. There were five patients who had no benefit from the vent because of blockage within the epitympanum or middle ear effusion. CONCLUSION: The Yung percutaneous mastoid vent can maintain long term patency. It is effective in the treatment of complete atelectasis as an adjunct to tympanoplasty. PMID- 15547417 TI - A new L-shaped titanium prosthesis for total reconstruction of the ossicular chain. AB - OBJECTIVE: To describe the technique used for total ossiculoplasty with the Fisch titanium total prosthesis and evaluate the 1-year postoperative functional results in patients presenting with the stapes (or footplate) without the malleus handle. STUDY DESIGN: Prospective trial with preoperative and postoperative comparison. SETTING: Academic and private practice tertiary care center for otologic surgery. PATIENTS: Forty-nine consecutive patients operated on from September 1996 to December 2000. SURGERY: : Staged ossicular reconstruction with a Fisch titanium total prosthesis placed between the footplate and the tympanic membrane without regard to the presence or absence of the stapes arch. Coupling of the prosthesis to the footplate was achieved by various techniques, including perforation, foot and spike on the footplate without perforation, and shaft alone (without foot) with tragal cartilage fixation (disc or small wedges). There was no interposition of cartilage between the prosthesis head and the tympanic membrane. MAIN OUTCOME MEASURES: Pre- and postoperative air and bone-conduction thresholds and air-bone gaps for pure-tone averages of three and four frequencies and for single frequencies. RESULTS: Postoperative air-bone gap closures within 20 dB distributed equally (50%) between 0.5, 1, and 4 kHz and reached the highest rate (89%) at 2 kHz (p < 0.05). The postoperative air-bone gaps for pure-tone averages reached 0 to 20 dB in 57% and 0 to 30 dB in 87% of the cases. There were no dead ears and no partial or total extrusions of prostheses. The best functional results were achieved through perforation coupling of the spiked foot to the footplate in large oval windows and after fixation of the shaft (without foot) with tragal cartilage disc in narrow oval windows. CONCLUSION: The functional results of the L-shaped Fisch titanium total prosthesis implanted in ears with the stapes but no malleus handle are best at 2 kHz and better than those of comparable columellar titanium prostheses over the remaining tested frequencies. PMID- 15547418 TI - Comparison of the mechanical performance of ossiculoplasty using a prosthetic malleus-to-stapes head with a tympanic membrane-to-stapes head assembly in a human cadaveric middle ear model. AB - HYPOTHESIS: Ossiculoplasty using prosthetic reconstruction with a malleus assembly to the stapes head will result in better transmission of vibrations from the eardrum to the stapes footplate than reconstruction with a tympanic membrane assembly to the stapes head. Both types of reconstruction will be affected by tension of the prosthesis. BACKGROUND: Theories (and some clinical studies) that the shape of the normal tympanic membrane is important suggest that prosthetic reconstruction to the malleus performs better than reconstruction to the tympanic membrane. This has not been previously tested by directly measuring vibration responses in the human ear. Our previous work suggests that tympanic membrane assembly to the stapes head type prostheses performed best under low tension. This had not been previously tested for malleus assembly to the stapes head type prostheses. METHODS: Hydroxyapatite prostheses were used to reconstruct a missing incus defect in a fresh cadaveric human ear model. Two types of prostheses were used, one from the stapes head to the malleus (malleus assembly to the stapes head), the other from the stapes head to the tympanic membrane (tympanic membrane assembly to the stapes head). Stapes footplate center responses were measured using a laser Doppler vibrometer in response to calibrated acoustic frequency sweeps. RESULTS: Tension had a very significant effect on both types of prostheses in the lower frequencies. Loose tension was best overall. The malleus assembly to the stapes head type prostheses consistently performed better than the tympanic membrane assembly to the stapes head type prostheses when stratified for tension. CONCLUSION: Tension has a significant effect on prosthesis function. Malleus assembly to the stapes head type prostheses generally result in better transmission of vibrations to the stapes footplate than tympanic membrane assembly to the stapes head type prostheses. PMID- 15547419 TI - Prevention of cisplatin ototoxicity using transtympanic N-acetylcysteine and lactate. AB - HYPOTHESIS: Transtympanic administration of the antioxidant N-acetylcysteine or lactated Ringer's solution onto the round window membrane will prevent cisplatin ototoxicity in the guinea pig model. BACKGROUND: Cochlear ototoxicity is a well known side effect of cisplatin administration, with the mechanism of injury thought to rest in oxidative damage to the outer hair cells. However, previous attempts at transtympanic antioxidant delivery have met with varied success. We present an effective method of counteracting cisplatin ototoxicity via the transtympanic application of lactated Ringer's solution or N-acetylcysteine. METHODS: Baseline distortion product otoacoustic emission measurements were obtained. Intraperitoneal cisplatin was administered to a cumulative dose of 20 mg/kg. The middle ears were either untreated (control) or filled with normal saline (negative control), 2%N-acetylcysteine diluted in normal saline (treatment), or lactated Ringer's solution (treatment) via anterosuperior quadrant myringotomies. Posttreatment distortion product otoacoustic emissions were obtained. RESULTS: Animals in the untreated control group and the negative control normal saline group demonstrated consistent obliteration of distortion product otoacoustic emissions. However, those receiving either lactated Ringer's solution or 2%N-acetylcysteine diluted in normal saline demonstrated significant preservation of distortion product otoacoustic emissions. The treatment regimen was well tolerated, with minimal animal loss. CONCLUSION: We have demonstrated the efficacy of transtympanic lactated Ringer's solution and N-acetylcysteine in the prevention of cisplatin ototoxicity using a guinea pig model. The possible mechanisms for the high efficacy of lactated Ringer's solution are discussed in detail. PMID- 15547420 TI - Usefulness of high doses of glucocorticoids and hyperbaric oxygen therapy in sudden sensorineural hearing loss treatment. AB - OBJECTIVE: We investigated the effect of pharmacologic (steroids, vasodilators, vitamins, and Betaserc) and hyperbaric oxygen therapy on patients with sudden sensorineural hearing loss. METHODS: The pharmacologic arm of the study consisted of 52 patients with defined sudden sensorineural hearing loss treated simultaneously in the ENT Department and National Center for Hyperbaric Medicine of the Medical University of Gdansk, Poland, from 1997 to 2000 (Group A). The hyperbaric oxygen therapy consisted of exposure to 100% oxygen at a pressure of 250 kPa for a total of 60 minutes in a multiplace hyperbaric chamber. The control group included 81 patients with defined sudden sensorineural hearing loss treated in the ENT Department, Medical University of Gdansk, from 1980 to 1996 (Group B). Both groups were comparable regarding the age of the patients, season of hearing loss occurrence, tinnitus and vestibular symptom frequency, delay before therapy, and average threshold loss before the start of treatment. The treatment results (hearing gain) were estimated using pure-tone audiometry. We retrospectively analyzed the audiograms of all patients. RESULTS: Patients from Group A (blood flow-promoting drugs, glucocorticoids in high doses, betahistine, and hyperbaric oxygen therapy) showed significantly better recovery of hearing levels compared with those from Group B (blood flow-promoting drugs and glucocorticoids in low doses) at seven frequencies (500, 1,000, 2,000, 3,000, 4,000, 6,000, and 8,000 Hz) (p < 0.05) and four groups of frequencies (pure-tone average, high-tone average, pure middle-tone average, and overall average) (p < 0.05). Percentage hearing gain in all investigated frequencies was also better in Group A versus Group B, and the differences were statistically significant (p < 0.05). CONCLUSION: We conclude that hyperbaric oxygen therapy with high doses of glucocorticoids improves the results of conventional sudden sensorineural hearing loss treatment and should be recommended. In addition, the best results are achieved if the treatment is started as early as possible. PMID- 15547421 TI - Normal hearing and language development in a deaf-born child. AB - BACKGROUND: Congenital deafness leads to major problems in speech, language, education, and social integration. Neonatal hearing screening and cochlear implantation now allow early hearing restoration. This article reports on a prospective longitudinal study of the first infant ever who received two cochlear implants in the prelexical period of her life. METHODS: The first deaf-born girl ever who received two implants at the ages of 5 and 15 months, respectively, was followed-up with repeated and detailed quantitative assessments from birth to 4 years of age. This consisted of 1) audiologic evaluation (audiometry, speech audiometry, and Categories of Auditory Performance score), 2) linguistic evaluation (monthly video analyses and tests of vocabulary, language skills, grammar, and intelligibility of the child's speech), and 3) descriptive assessment of the educational setting. RESULTS: All results lie within the 95% confidence interval of hearing peers. The audiologic performance lies at or above average from age 2 years onward. The child started babbling at the normal age of 8 months. Her linguistic skills increased from low percentiles before age 2 to above average from age 2 for comprehension and from age 3 for production. The grammar and intelligibility of the child's speech increased from low percentiles to average at age 4. The girl entered preschool at the normal age of 2.5 years, and this with only very limited special assistance. CONCLUSION: This case illustrates the fact that congenital deafness no longer has to lead to abnormal hearing and abnormal speech development. It opens the debate of the ethics of not implanting a deaf child in the first few months of life. PMID- 15547422 TI - Auditory perception and speech discrimination after cochlear implantation in patients with connexin 26 (GJB2) gene-related deafness. AB - HYPOTHESIS: Auditory perception and speech discrimination among pediatric cochlear implantees may vary because of underlying deafness etiology, including connexin 26 (GJB2) gene-related deafness. BACKGROUND: Preliminary data suggest pathologic changes due to GJB2 mutations do not affect the spiral ganglion cells, which are stimulated by the cochlear implant. The survival of the spiral ganglion cells is believed to be an important determinant of outcome after surgery. Patients with GJB2-related deafness may therefore have enhanced prospects for good speech discrimination after implantation. METHODS: In an observational cohort study, GJB2 mutation analysis was performed using polymerase chain reaction amplification and direct sequencing on 31 prelingually deaf pediatric cochlear implantees, of which there were 30 with nonsyndromic deafness of unknown etiology, and one with keratitis-ichthyosis-deafness syndrome. Speech discrmination was assessed prospectively when they had reached postoperative year 3 using the IOWA Matrix Level B Sentences test and Glendonald Auditory Screening Procedure (GASP), with both patients and assessors blind to GJB2 status. RESULTS: Eleven patients had GJB2-related deafness and 20 patients had GJB2-unrelated deafness. IOWA Matrix scores were higher in patients with GJB2-related deafness but did not reach statistical significance. However, GASP scores were statistically significantly higher in patients with GJB2-related deafness (median word score, 92%; median sentence score, 80%), compared with those of patients with GJB2-unrelated deafness (median word score, 63%; median sentence score, 45%; word score, p = 0.037; sentence score, p = 0.045). Ordinal logistic regression analysis on IOWA Matrix and GASP sentence scores found better statistically significant scores in patients with GJB2-related deafness (p < 0.05) after adjustment for confounding variables. CONCLUSION: Pediatric cochlear implantees with GJB2-related deafness appear to have equal or better speech discrimination compared with a group of prelingually deaf children with deafness of unknown etiology. PMID- 15547423 TI - Connexin 26 (GJB2) gene-related deafness and speech intelligibility after cochlear implantation. AB - HYPOTHESIS: Speech intelligibility in children after cochlear implantation may depend on their deafness cause, including connexin 26 (GJB2) gene-related deafness. BACKGROUND: There is significant variability in the degree of intelligibility, or clarity, of children's speech after cochlear implantation. GJB2 gene-related deafness may be a factor, as preliminary data suggest that pathologic changes do not affect the spiral ganglion cells, which are the neural elements stimulated by the implant, thus favoring better results. METHODS: In an observational retrospective cohort study of pediatric cochlear implantees, 38 patients with nonsyndromic deafness of unknown cause and 1 with keratitisichthyosis-deafness syndrome underwent GJB2 mutation analysis using polymerase chain reaction amplification and direct sequencing. The primary outcome measure assessed was Speech Intelligibility Rating score from postoperative Year 1 (n = 39) to Year 5 (n = 17). Educational setting was considered as a secondary outcome measure. Statistical analysis was double blinded, with patients and assessors of outcome unaware of GJB2 status. RESULTS: Fourteen patients had GJB2-related deafness and 25 had GJB2-unrelated deafness. Comparisons at Year 3 (n = 31) revealed intelligible speech achieved by 9 of 11 with GJB2-related deafness, compared with only 6 of 20 with GJB2-unrelated deafness (p = 0.017). Ordinal logistic regression analysis on Speech Intelligibility Rating scores found statistically significantly better scores in children with GJB2-related deafness (p < 0.05) both before and after adjustment for confounding variables. A larger proportion with GJB2-related deafness also attended mainstream school (p = 0.01). CONCLUSION: In pediatric cochlear implantees, GJB2-related deafness is a predictor of good speech intelligibility. PMID- 15547424 TI - Cochlear implantation in 53 patients with otosclerosis: demographics, computed tomographic scanning, surgery, and complications. AB - OBJECTIVES: To collect data from a large number of cochlear implant recipients with otosclerosis and to make an assessment of these patients' clinical characteristics, computed tomographic scans, surgical findings, and complications, and to quantify the occurrence of postoperative facial nerve stimulation. STUDY DESIGN: Retrospective multicenter study. PATIENTS: Fifty-three patients with otosclerosis from four cochlear implant centers in the United Kingdom and The Netherlands were reviewed. Sixty surgical procedures were performed in these patients: 57 devices were placed in 56 ears. RESULTS: The computed tomographic imaging demonstrated retrofenestral (cochlear) otosclerotic lesions in the majority of patients. Although not statistically significant, the extent of otosclerotic lesions on the computed tomographic scan as categorized in three types tends to be greater in patients with rapidly progressive hearing loss, in patients in whom there is surgically problematic insertion of the electrode array, and in patients with facial nerve stimulation. In four patients, revision surgery had to be performed. Twenty of 53 (38%) patients experienced facial nerve stimulation at various periods postoperatively. CONCLUSION: Cochlear implant surgery in patients with otosclerosis can be challenging, with a relatively high number of partial insertions and misplacements of the electrode array demanding revision surgery. A very high proportion of patients experienced facial nerve stimulation mainly caused by the distal electrodes. This must be discussed with patients preoperatively. PMID- 15547425 TI - Bacterial biofilms may contribute to persistent cochlear implant infection. AB - OBJECTIVES: To determine if bacterial biofilms are present on the surface of extruding or persistently infected cochlear implants. METHODS: Scanning electron microscopy was performed on cochlear implants removed from two patients because of recalcitrant infection, two implants removed secondary to device failure, and two devices that had never been implanted. Preparations were examined by experienced microbiologists for the presence of bacterial biofilms. RESULTS: Microorganisms and amorphous extracellular debris were found on the surface of the infected cochlear implants and the implants removed because of device failure. Biofilm formation was deemed definite in one infected device and possible in the other explanted devices. The never-implanted controls demonstrated microbial contamination without exopolymeric matrix, inconsistent with biofilms. CONCLUSION: Bacterial biofilm formation may play a role in recalcitrant cochlear implant infections. This may have profound implications for the treatment of cochlear implant infections. PMID- 15547426 TI - Benefits of bilateral electrical stimulation with the nucleus cochlear implant in adults: 6-month postoperative results. AB - OBJECTIVE: To evaluate the benefits of bilateral electrical stimulation for hearing-impaired adult subjects using the Nucleus 24 cochlear implant in a multicenter study, and to compare and quantify performance on speech perception measures in quiet and in noise and localization ability for unilateral and bilateral cochlear implant use. DESIGN: : Repeated single subject measures were carried out for each subject, with each subject serving as their own control. Assessment of unilateral and bilateral listening conditions for performance on tests of speech comprehension and sound localization were performed. Speech comprehension measures were performed in quiet at 0 degree azimuth and in the presence of background noise simultaneously presented from the same speaker and spatially separated by 90 degrees, at S+45 degrees N45 degrees and at S-45 degrees N+45 degrees . Test materials included Freiburger monosyllabic words, Oldenburger sentences, and the Hochmair-Schulz-Moser sentences. Tests of localization were performed in the horizontal plane with 12 speaker locations 30 degrees apart using a shortened sentence stimulus from the Hochmair-Schulz-Moser sentences at two possible presentation levels of 55 and 70 dB sound pressure level for assessment of directionality. The binaural advantage provided by bilateral stimulation was calculated with respect to each ear separately, classified as either the better or poorer performing ear for each speech material in quiet and in noise test conditions. For localization of sound, the binaural advantage was compared with left and right ears separately. Paired comparisons for performance data in all conditions were carried out by considering measurements for each subject in different conditions as paired observations and applying the Student's t test to determine the statistical difference between the data sets. SETTING: Tertiary referral centers with a cochlear implant program. PATIENTS: Thirty-seven profoundly hearing-impaired adults were enrolled in the study, 22 simultaneously and 15 sequentially bilaterally implanted. All patients received the Nucleus 24 cochlear implant and used the Nucleus SPrint or ESPrit 3G speech processor, with the vast majority using the ACE speech coding strategy. RESULTS: For spatially separated speech in noise conditions, an interaural performance advantage for the ear closest to the speech source (i.e., with a superior signal to noise ratio) compared with that for the ear closest to the noise source (i.e., with an inferior signal to noise ratio) is consistently demonstrated regardless of whether it is the better or poorer performing ear closest to the speech signal. This is referred to as a significant binaural head shadow benefit, resulting in a mean improvement between -10 dB and -11.4 dB in the critical signal to noise ratio required for 50% speech comprehension for the Olden-burger sentences and a mean improvement in the maximum score of 42% to 55% for the ear closest to the speech signal over the ear farthest away for the Hochmair-Schulz-Moser sentences. Bilateral stimulation is always observed to provide a performance advantage over the unilateral listening condition for either ear when ipsilateral to the noise source. In addition, as demonstrated by approximately half the subjects tested in noise with the Hochmair-Schulz-Moser sentences, a performance advantage of bilateral stimulation may be observed over the better ear alone when positioned ipsilateral to the speech signal, which is referred to as a binaural squelch effect. On average, for the group, this resulted in a statistically significant improvement in speech comprehension scores of 8% in the bilateral listening condition compared with the scores for the better ear alone. Through assessment of comprehension of coincidental speech in noise and speech in quiet, a significant benefit of binaural redundancy was noted for the group for Oldenburger sentence scores in noise and in quiet compared with unilateral scores for either ear and for the Freiburger monosyllabic words in quiet in comparison with the better ear alone scores. Binaural stimulation also led to a significant improvement in localization ability over either monaural condition, with the root mean square degrees of error reduced by 38 degrees compared with that observed for unilateral stimulation. CONCLUSION: Similar to what has been observed for bilateral acoustic stimulation in the past, bilateral electrical stimulation provides the foundation for the potential advantages of the head-shadow effect, providing a binaural head shadow benefit and binaural auditory processing such as binaural redundancy and binaural squelch effects, all of which combine to lead to improved speech comprehension over unilateral listening conditions. The combination of improved speech comprehension and improved localization ability made available through bilateral electrical stimulation provides the necessary foundation to further assist the hearing-impaired listener to better cope with communication in the everyday listening situation both in noise and in quiet. PMID- 15547427 TI - MRI scanning and incus fixation in vibrant soundbridge implantation. AB - OBJECTIVE: Presentation of patient data after Vibrant Sound-bridge (VS) implantation/explantation with respect to magnetic resonance imaging (MRI) compatibility and stability of incus fixation of the implant. STUDY DESIGN: In a retrospective case review, we report on two patients who had to be diagnosed by cranial MRI scans after previous implantation of a VS. Moreover, in one of these cases, the incus was removed after explantation of the VS because of a peripheral hearing loss. This enabled a microscopic evaluation of the incus. At the long process of the incus, the floating mass transmitter (FMT) has been fixed for 4 years. SETTING: Tertiary referral center. SUBJECTS: Two patients who fulfilled, at the time of implantation, the criteria for implantation of a VS. RESULTS: In both cases, no demagnetization of the external magnet nor of the FMT were found. Device function after the MRI scans were normal. Pure-tone audiometric thresholds remained unchanged after the MRI scanning and the scanning-related magnetic forces. In both cases, the FMT fixation of the incus was found to be regular. In one case, the fixation was checked by an additional tympanoscopy 1 year after the implantation, and in the other case it was assessed by recording the functional gain of the VS (which was normal) after the MRI scanning. One patient complained about a transient hyperacuity due to the loudness during the MRI scanning procedure. MRI scans showed a blackening of the implant area with spherical distortions of the picture. CONCLUSION: Although MRI scanning (at 1.5 tesla [T]) with the FMT in place did not lead to adverse effects in the two patients, systematic in vitro studies are required to determine a possible magnetization threshold that could impair the VS function when MRI scans are applied in those patients. The microscopically observed erosions of the long process of the incus after 4 years of FMT clamp fixation show similarities to findings after stapes revision surgery. However, this limited experience in one case does not allow us to make conclusions on the long-term stability of the incus fixation. PMID- 15547428 TI - Transcanal anterior approach for cystic lesions of the petrous apex. AB - OBJECTIVE: To investigate the utility of a newly described approach, the transcanal anterior approach that is a modification of the subcochlear approach for the drainage of cystic lesions of the petrous apex. STUDY DESIGN: Prospective temporal bone study. SETTING: Tertiary referral center. MATERIALS: A total of six cadaveric temporal bone specimens (four males) were included. Right ear in four specimens and left ear in two specimens were used. INTERVENTIONS: The approach commenced with postauricular skin incision. After the transsection of the meatal skin, antero-inferior tympanotomy was performed. Anteroinferior canaloplasty localized the carotid canal. As much as 0.5 cm of the vertical segment of the internal carotid artery was skeletonized. After the identification of the artery, petrous apex cells were reached by drilling the cortical bone between the cochlea and the internal carotid artery. An air cell tract was established. Position and length of the tract in two specimens were demonstrated on the 1 mm-cut computerized tomography scans. MAIN OUTCOME MEASURES: Utility of the approach was investigated based on the established criteria: anteroposterior diameter and height of the fenestra of the tract, length of the tract from the cochlea to the deepest point of the tract (depth of the tract), and the injury risk of the internal carotid artery and the cochlea. RESULTS: The mean anteroposterior diameter, the height, and the length were 4.7 +/- 1.05 mm, 3.2 +/- 0.68 mm, and 14.7 +/- 1.1 mm, respectively. Injury did not occur in the cochlea or internal carotid artery in any of the specimens. CONCLUSION: With the subcochlear approach, there is always potential risk of injury to the cochlea, the internal carotid artery, and the jugular bulb. Although this new approach includes a technically challenging stage (exposing the vertical portion of the internal carotid artery), partly exposing the vertical portion of the artery provides a safer approach, which decreases the injury risk for the round window and the jugular bulb. Additionally, measurements show that it is possible to reach a considerable part of the petrous apex cells. PMID- 15547429 TI - Aging effect on vestibular evoked myogenic potential. AB - OBJECTIVE: Vestibular evoked myogenic potential (VEMP) is applied to explore the integrity of sacculocollic reflex. Although tests to evaluate vestibular-ocular reflex pathway have shown that vestibular function is adversely affected by aging, VEMP, in this study, is used as a novel test to define how aging influences sacculocollic reflex pathway. STUDY DESIGN: Prospective study. SETTING: Academic tertiary referral center. SUBJECTS: Eighty normal subjects, equally divided into four groups according to their age, were enrolled to this study. Group I included patients aged <20 years, Group II patient ages ranged from 21 to 40 years, Group III patients were 41 to 60 years, and Group IV included patients older than 60 years. INTERVENTIONS: Recordings of VEMP responses. MAIN OUTCOME MEASURES: The response rate and parameters of VEMP, including p13 latency, n23 latency, amplitude, and interaural difference ratio. RESULTS: The VEMP response rates from Groups I to IV was 98%, 98%, 90%, and 60%, respectively, disclosing a significant difference only between Group IV and other groups (p < 0.05). The amplitude was negatively correlated with age in contrast to the n23 latency, correlating positively with age; both reached a significant difference (p < 0.05). Although the p13 latency had a trend to prolong as age increased, no significant correlation existed (p < 0.06). Moreover, the interaural difference ratio was also not significantly correlated with age. CONCLUSIONS: As age increased over 60 years, the VEMP response rate decreased dramatically. While age increased, the VEMP amplitude decreased in comparison to n23 latency prolonged. These findings might suggest that aging could deteriorate the saccular and corresponding neural functions. When interpreting the VEMP parameters, it should be kept in mind that aging could affect VEMP responses. Based on this study, we suggest establishing different reference values according to different age groups when evaluating VEMP response in patients with vestibular diseases. PMID- 15547430 TI - Vestibular evoked myogenic potentials versus vestibular test battery in patients with Meniere's disease. AB - OBJECTIVE: The present study was undertaken to assess the sensitivity of vestibular evoked myogenic potentials testing to side-of-disease in Meniere's disease patients and to test the hypothesis that information supplied by vestibular evoked myogenic potentials is complementary to that provided by a conventional vestibular test battery. STUDY DESIGN: Prospective cohort study. SETTING: Large specialty hospital, department of otolaryngology. SUBJECTS: Twenty consenting adults (9 men and 11 women) with unilateral Meniere's disease by American Academy of Otolaryngology-Head and Neck Surgery diagnostic criteria. INTERVENTIONS: All subjects underwent bilateral vestibular evoked myogenic potentials testing using ipsilateral broadband click and short-toneburst stimuli at 250, 500, and 1,000 Hz. All subjects also underwent electronystagmography and sinusoidal vertical axis rotation testing. MAIN OUTCOME MEASURES: Accuracy of side-of-disease assignment by vestibular evoked myogenic potentials, caloric asymmetry, and multivariate analysis. RESULTS: Side-of-disease assignment was most accurate using caloric asymmetry with a 5% interaural difference criterion, achieving 85% correct assignment. The next best method was vestibular evoked myogenic potentials using 250-Hz toneburst stimuli, achieving 80% correct assignment. The least accurate method was caloric asymmetry using a traditional 30% interaural difference limen, achieving 55% correct assignment. Comparison of 5% interaural difference criterion and vestibular evoked myogenic potentials using 250-Hz toneburst stimuli showed discordant results, but in no case did both 5% interaural difference criterion and vestibular evoked myogenic potentials using 250-Hz toneburst stimuli make an incorrect assignment. CONCLUSION: Vestibular evoked myogenic potentials threshold was shown to be highly sensitive to side-of-disease in unilateral Meniere's disease. We observed instances of discordance in side-of-disease assignment by caloric asymmetry and vestibular evoked myogenic potential methods but no case in which both methods were incorrect. This supports the hypothesis that vestibular evoked myogenic potentials supplies information complementary to that provided by other components of the vestibular test battery. PMID- 15547431 TI - Audiometric findings of patients with migraine-associated dizziness. AB - OBJECTIVE: The objective of this study was to determine whether the audiometric findings of migraine-associated dizziness could be used to better distinguish migraine-associated dizziness from Meniere's disease. STUDY DESIGN: A retrospective chart review. SETTING: Tertiary, otology/neurotology practice. PATIENTS: Two groups of patients were studied, a migraine-associated dizziness and a Meniere's disease group. There were 76 and 34 patients in the migraine associated dizziness and Meniere's disease groups, respectively. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Initial and follow-up pure-tone average and low frequency pure-tone average were recorded for both groups. Independent samples t tests were used to test for mean differences in pure-tone average and low frequency pure-tone average. RESULTS: Pure-tone average and low-frequency pure tone average were significantly worse for patients in the Meniere's disease group at both the initial and follow-up assessments. Three patients in the migraine associated dizziness group had an elevated pure-tone average (>/=26 dB) and/or low-frequency pure-tone average at initial and/or follow-up assessment. The remaining 73 migraine-associated dizziness patients had normal hearing. In the Meniere's disease group, only two patients had a normal pure-tone average and low frequency pure-tone average at both initial and follow-up evaluations. The hearing difference between the two groups was significant even when controlling for age and duration of dizziness symptoms. CONCLUSION: Audiometric findings of patients with migraine-associated dizziness are most often normal. Unlike Meniere's disease, the sensorineural hearing loss in migraine-associated dizziness rarely progresses. These audiometric findings may help to distinguish migraine-associated dizziness from Meniere's disease when diagnostic ambiguity exists between these two diagnoses. PMID- 15547432 TI - Subarcuate venous malformation causing audio-vestibular symptoms similar to those in superior canal dehiscence syndrome. AB - OBJECTIVE: To present a patient with symptoms similar to those of superior canal dehiscence syndrome due to another cause. STUDY DESIGN: Case report. SETTING: University hospital, tertiary referral center. PATIENT: The 65-year-old woman had suffered for 4 years from hearing loss, tinnitus, and pressure-induced vertigo. INTERVENTION: Audio-vestibular testing, high-resolution computed tomography, and magnetic resonance angiography. MAIN OUTCOME MEASURE: The superior canal dehiscence syndrome is caused by failure of normal postnatal bone development in the middle cranial fossa leading to absence of bone at the most superior part of the superior semicircular canal. The typical features for this syndrome are sound and pressure-induced vertigo with torsional eye movements, pulse synchronous tinnitus and apparent conductive hearing loss in spite of normal middle ear function. We present a patient with very similar symptoms and findings, who instead had a superior canal dehiscence close to the common crus. Neuroradiologic findings suggested that the dehiscence was related to a venous malformation. CONCLUSIONS: Symptoms and findings suggesting superior canal dehiscence syndrome can have a different cause. PMID- 15547433 TI - Management of the internal carotid artery in tumors of the lateral skull base: preoperative permanent balloon occlusion without reconstruction. AB - OBJECTIVE: To present our experience with permanent preoperative balloon occlusion of the internal carotid artery while dealing with different abnormalities of the lateral skull base and a comparison with the results mentioned in the literature. STUDY DESIGN: Retrospective case review. SETTING: Private neurotologic and skull base tertiary referral center. PATIENTS: Fifteen patients who underwent preoperative balloon occlusion of the internal carotid artery and surgery subsequently for various abnormalities of the lateral skull base between 1989 and 2002. INTERVENTIONS: Each patient was subjected to four vessel angiography along with the manual cross-compression test and balloon test occlusion to assess the efficacy of the collateral circulation. After angiography, each patient underwent a preoperative balloon occlusion, after which a lateral skull base procedure was performed for removal of the abnormality. MAIN OUTCOME MEASURES: Only those patients showing evidence of adequate collateral cerebral circulation and a less than 1-second delay between the angiographic phases of the two cerebral hemispheres on angiography were considered fit for preoperative balloon occlusion. While under going the preoperative balloon occlusion, the patients were clinically assessed for the development of any neurologic symptoms and signs. Long-term follow-up after surgery was also based on the development of symptoms and signs of neurovascular compromise. RESULTS: A major complication in the form of long-lasting hemiplegia occurred in one patient (6.7%). This complication was the result of technical factors rather than an effect of cerebral ischemia, because it was caused by an intimal dissection produced by the catheter. A defect in the visual field occurred in one patient (6.7%) that resolved partially after antiplatelet therapy. There was no mortality in our series related to preoperative balloon occlusion of the internal carotid artery. CONCLUSION: Preoperative balloon occlusion of the internal carotid artery can still be considered a viable option for the management of the internal carotid artery during lateral skull base surgery. Proper preoperative evaluation of the adequacy and efficacy of the collateral cerebral circulation reduces the chances of postoperative neurovascular complications. PMID- 15547434 TI - T-cell prolymphocytic leukemia presenting as an ear canal mass. AB - OBJECTIVE: We discuss the first reported case of a chronic leukemia involving the external auditory canal. STUDY DESIGN: Case report and review of the literature (Medline, 1962-2003). METHODS AND RESULTS: An adult man in good health came to receive medical attention because of a sensation of aural fullness. Radiologic and physical examination demonstrated an ovoid soft tissue mass of the lateral external auditory canal that, after biopsy and flow cytometry studies, was found to be T-cell prolymphocytic leukemia. The patient presented with minimal disease in a highly symptomatic location, allowing early initiation of therapy and extending his life by nearly four times the average life expectancy for this aggressive chronic leukemia. CONCLUSION: Lymphomas of the external auditory canal are extremely rare neoplasms. Only a few cases have previously been reported, and all of the previous reports involve acute leukemias. This report indicates that chronic leukemias can present with otologic manifestations such as an external auditory canal mass. PMID- 15547435 TI - Gunshot wounds to the facial nerve. AB - OBJECTIVE: This paper presents our experience with gunshot wounds to the temporal bone and discusses facial nerve lesions, surgical indication, surgical timing, and other findings. STUDY DESIGN: We performed a retrospective review of patients treated for facial nerve lesion after gunshot injury to the temporal bone. SETTING: The study was performed in the Otolaryngology Department of the University of Sao Paulo Medical School, Sao Paulo, Brazil. PATIENTS: Ninety-eight patients treated between 1988 and 1999 were analyzed. INTERVENTION: Facial nerve lesions, bullet locations, and surgical techniques were analyzed. Patients were monitored for 2 years. RESULTS: Gunshot trauma to the temporal bone presented considerable tissue loss resulting from the abrasion effect and severity of the impact. The third segment of the facial nerve was most affected, and the bullet was typically found lodged in the mastoid tip. Postoperative infection was common. Such cases required revision surgery, resulting in worse cosmetic outcomes than in cases of closed trauma. CONCLUSIONS: Surgical exploration of the facial nerve should be performed as soon as possible, since long delays increase the chance of traumatic neuroma and more pronounced scarring around the facial nerve. Open mastoidectomy with meatoplasty is the surgical technique recommended for repairing the mastoid and the facial nerve. In the majority of cases, a cable graft is necessary. Since nerve lesion in proximity to the stylomastoid foramen and extratemporal facial nerve is common, these areas must be explored carefully. PMID- 15547436 TI - Expression of emotion and quality of life after facial nerve paralysis. AB - OBJECTIVE: To investigate the facial expression of emotion and quality of life in patients after long-term facial nerve paralysis. STUDY DESIGN: Cross-sectional. SETTING: Facial nerve paralysis clinic. PATIENTS: Twenty-four patients with facial nerve paralysis and 24 significant others (partner, relative, friend). INTERVENTION: Patients were assessed using Sunnybrook, Sydney, and House Brackmann grading scales and SF-36, Glasgow Benefit Inventory, and Facial Disability Index quality-of-life measures. RESULTS: When patients identified themselves as either effective or not effective at facially communicating each of Ekman's primary emotions (happiness, disgust, surprise, anger, sadness, and fear), 50% classified themselves as not effective at expressing one or more of the six emotions. Significant others of the not effective patients rated the emotions as more difficult for their partner-patients to communicate facially than did the significant others of effective patients. The SF-36 quality-of-life survey revealed lower social functioning relative to physical functioning for not effective patients. From the Sunnybrook Facial Grading System, more synkinesis was found for those patients not effective at expressing happiness, less brow and eye movement for patients not effective at expressing sadness, and less voluntary movement for those not effective with surprise. CONCLUSION: Movement deficits associated with expressing specific emotions and an association with quality-of life measures were identified in patients with long-term facial nerve paralysis who saw themselves as not effective at facial expression of emotions. To improve management of emotional expression in patients with facial nerve paralysis, a broader approach is recommended, linking the practitioner's treatment goals with patient-driven outcome goals. PMID- 15547437 TI - Agreement between the Sunnybrook, House-Brackmann, and Yanagihara facial nerve grading systems in Bell's palsy. AB - OBJECTIVE: To assess the agreement between the Sunnybrook facial nerve grading system and the House-Brackmann and Yanagihara systems. STUDY DESIGN: Prospective clinical facial nerve grading. SETTING: Tertiary referral center. PATIENTS: One hundred assessments, 94 in patients with Bell's palsy and 6 with herpes zoster. INTERVENTION: Diagnostic. MAIN OUTCOME MEASURES: Evaluation according to the weighted regional Sunnybrook system, the gross House-Brackmann system, and the unweighted regional Yanagihara system. Weighted kappa statistics was used to measure agreement between the grading systems. RESULTS: The average weighted kappa value between the Sunny-brook, House-Brackmann, and Yanagihara grading systems was 0.65; kappa values increased temporally (but not statistically significantly) up to day 180. The highest agreement value, 0.72, was found between the Sunnybrook and Yanagihara grading systems. The weighted kappa value between the Sunnybrook and House-Brackmann systems was 0.59. In Sunnybrook gradings less than 63, there was an overlap between House-Brackmann scores of III to VI. Reliable conversion tables between the gross House-Brackmann system and the regional Sunnybrook and Yanagihara systems could not be established. CONCLUSION: The Sunnybrook system scores at the same agreement level as the House Brackmann and Yanagihara grading systems. There is an evaluative difference between the weighted regional Sunnybrook and the gross House-Brackmann systems. Substantial agreement was found between the regional Sunnybrook and Yanagihara scales. Sunnybrook grading is easy and quick. By adding objective measurements and additional secondary defects, the Sunnybrook system can be an alternative to the other predominating grading systems. PMID- 15547438 TI - Lipoma of the internal auditory canal and cerebellopontine angle. PMID- 15547439 TI - Cochlear endosteal erosion with focal osteomyelitis induced by cochlear implantation. PMID- 15547440 TI - Passive and dynamic properties of the eustachian tube: quantitative studies in a model. AB - OBJECTIVE: To question the conception of Eustachian tube (ET) dysfunction by investigating some of its passive and active properties. BACKGROUND: Middle ear (ME) fluid accumulation accompanied by a negative pressure is generally accepted to be induced by ET dysfunction, implying that the origin of pathology is within the tube. METHODS: Pressure changes were continuously and directly measured in a ME model having human dimensions, constructed with a patent ET, and employing different mastoid sizes. Various liquids were introduced into the ME chamber while maintaining atmospheric pressure. The model was tilted to a 45 degrees angle below the horizon, causing the liquid to block the tympanic inlet of the ET. Then either the ME was opened to the atmosphere (simulating an insertion of a ventilating tube) and the ET was inspected for liquid passage through it, or the cartilaginous ET was compressed and opened, imitating the "pumping" action of the ET at the process of deglutition. RESULTS: ET blockage produced a ME negative pressure in the range of -8 to -10 mm H2O. ET pumping caused a ME pressure change of 2 to 60 mm H2O, a greater change when a smaller mastoid was used. CONCLUSIONS: ET obstruction by intratympanic fluid may cause a negative pressure in an ME having a normal, patent ET. Clearance of such fluid would be facilitated according to its viscosity. The ET pumping mechanism may play a major role in fluid transport and pressure regulation of the ME. PMID- 15547441 TI - Intratympanic steroid treatment: a review. AB - OBJECTIVE: To review published literature regarding the use of intratympanic steroids in the treatment of Meniere's disease and sudden sensorineural hearing loss and to make recommendations regarding their use based on the literature review. DATA SOURCES: Literature review from 1996 to 2003, PubMed, Medline Plus, and Web of Science. STUDY SELECTION: Retrospective case series and uncontrolled prospective cohort studies were the only types of studies available for review. CONCLUSION: On the basis of the available literature, a weak recommendation is made to use intratympanic steroid treatment of sudden hearing loss if oral steroid therapy fails or is contraindicated. The available studies regarding intratympanic steroid treatment of Meniere's disease and tinnitus are inadequate to answer the question of the efficacy of this treatment for these conditions. Higher quality studies are needed. PMID- 15547443 TI - Intracranial extension of jugular foramen tumors. PMID- 15547444 TI - Let's not discourage innovative approaches to complex questions. PMID- 15547445 TI - Association study of brain-derived neurotrophic factor gene polymorphism and alcoholism. AB - OBJECTIVE: Brain-derived neurotrophic factor (BDNF) influences dopamine and serotonin neurotransmitters that are heavily linked to addiction. A quantitative trait loci study indicated that genes localized to 11p13, where the BDNF gene is mapped (11p13-15), increase the risk for severe alcohol withdrawal. Moreover, a recent study using a pooled-sample microarray suggested that the BDNF gene locus was included in the loci that were shown to be associated with drug abuse. These lines of evidence suggested that BDNF might play some role in the development of or vulnerability to alcoholism and/or clinical characteristics of alcoholic individuals. METHODS: The alcoholic subjects consisted of 377 male Japanese patients. A structured interview was used to obtain social background, drinking history, history of violence while intoxicated, history of alcohol withdrawal, and family history of alcoholism. The control group consisted of 336 nonalcoholic male subjects. Genotyping of the G196A polymorphism of the BDNF gene was done by polymerase chain reaction (PCR)-restriction fragment length polymorphism method. RESULTS: Genotype and allele distributions of the BDNF gene polymorphism did not differ significantly between alcoholic and control subjects. However, comparing clinical characteristics across G196A genotypes, we found that alcoholic subjects with violent tendencies and a history of delirium tremens had a significantly higher frequency of AA genotypes and A allele frequencies than those without them. Moreover, alcoholic subjects with the A allele had earlier onset of the disease than those without it. CONCLUSIONS: These results indicate that BDNF gene polymorphism might modify phenotypes of alcoholism. PMID- 15547446 TI - Confirmation and genetic dissection of a major quantitative trait locus for alcohol preference drinking. AB - OBJECTIVE: In previous work, we created congenic strains that carry the DBA/2IBG (D2) region for alcohol preference on chromosome 2, on an otherwise C57BL/6IBG (B6) background. Here, we report construction and testing of interval-specific congenic recombinant strains (ISCRSs) for the purpose of narrowing the quantitative trait loci (QTL) interval. METHODS: ISCRSs were derived by identifying mice that carry recombination events in the D2 interval, during the backcrossing for congenics. Recombinant mice were backcrossed to B6, and progeny that carry the reduced chromosome 2 region were tested for its effect on the alcohol preference phenotype. RESULTS: We developed multiple ISCR strains, which spanned the QTL interval. Three of these showed the D2 phenotype of reduced alcohol consumption. The overlap of two of these strains reduced the QTL interval from 66.8 to 3.5 Mb. A third positive ISCRS suggests the possibility of a second, linked QTL. CONCLUSIONS: Use of ISCRSs can narrow a QTL region to a few Mb. This reduced interval size will facilitate identification of candidate genes, through bioinformatics, gene expression, and DNA sequencing strategies. Potential difficulties, including reduced power as a result of variable phenotypes or small effect size, are discussed. PMID- 15547447 TI - Glutathione S-transferase 8-8 expression is lower in alcohol-preferring than in alcohol-nonpreferring rats. AB - OBJECTIVE: A primary focus of alcohol research is to provide novel targets for alcohol treatment by identifying genes that predispose individuals to drink alcohol. Animal models of alcoholism developed by selective breeding are invaluable tools to elucidate both the genetic nature and the underlying biological mechanisms that contribute to alcohol dependence. These selected lines (high alcohol preferring and low alcohol preferring) display phenotypic and genetic differences that can be studied to further our understanding of alcohol preference and related genetic traits. By combining molecular techniques, genetic and physiological factors that underlie the cause of alcoholism can be identified. METHODS: Total gene expression analysis was used to identify genes that are differentially expressed in specific brain regions between alcohol naive, inbred alcohol-preferring (iP) and -nonpreferring (iNP) rats. Quantitative reverse transcriptase-polymerase chain reaction, in situ hybridization, Western blot, and sequence analysis were used to further characterize rat glutathione S transferase 8-8 (rGST 8-8). RESULTS: Lower expression of rGST 8-8 mRNA was observed in discrete brain regions of iP compared with iNP animals, and these expression differences were confirmed. To determine additional expression patterns of rGST 8-8, we used in situ hybridization. Rat GST 8-8 was highly expressed in hippocampus, the choroid plexus of the dorsal third ventricle and the lateral ventricle, and ependymal cells along the dorsal third ventricle and the third ventricle. Western blot analysis showed that rGST 8-8 protein levels were lower in the hippocampus and the amygdala of iP compared with iNP. A silent single-nucleotide polymorphism in the coding region and three single-nucleotide polymorphisms in the 3'-UTR were identified in the rGST 8-8 cDNA. CONCLUSION: There is regional variation of rGST 8-8 expression in the brain, at both the mRNA and protein level, and the iP strain has lower innate rGST 8-8 levels than the iNP strain in discrete brain regions. PMID- 15547449 TI - Transfer of phosphatidylethanol between lipoproteins. AB - BACKGROUND: Phosphatidylethanol (PEth) is an abnormal phospholipid formed only in the presence of ethanol. It has been recently shown that lipoprotein-associated PEth may mediate the effects of ethanol on endothelial cells, and this may explain, at least in part, the beneficial effect of ethanol on atherosclerosis. This study was performed to investigate the transfer of PEth between lipoproteins and the effects of PEth on cholesteryl ester transfer protein (CETP) activity in plasma. METHODS: Lipoproteins were isolated from the plasma of healthy male volunteers (n = 16) and male alcoholics (n = 13). The transfer of cholesteryl esters and PEth was determined between labeled low-density lipoprotein (LDL) and unlabeled high-density lipoprotein particles in vitro. The electrophoretic mobility of PEth-modified LDL particles was determined by agarose gel electrophoresis. RESULTS: PEth was transferred from PEth-modified LDL to high density lipoprotein at an initial rate of 25.9 nmol/ml/hr. Monoclonal antibody (TP2) against the putative lipid-binding domain of CETP inhibited the transfer rate of PEth by approximately 64%, whereas the cholesteryl ester transfer was inhibited by 86%. This indicates that most of PEth was transferred by transfer proteins other than CETP. CONCLUSIONS: The transfer of PEth between lipoproteins enables the redistribution of PEth from lipoprotein fractions with a slow turnover to those with a rapid clearance. Moreover, the PEth-induced change in the electrical charge of lipoproteins may affect the binding of lipoproteins to their receptors and binding proteins. This in turn may alter the metabolism of lipoproteins and lipid-mediated signaling pathways in the cells delineating the vascular wall. PMID- 15547448 TI - Bitter receptor gene (TAS2R38), 6-n-propylthiouracil (PROP) bitterness and alcohol intake. AB - BACKGROUND: Phenylthiocarbamide (PTC) and 6-n-propylthiouracil (PROP), chemically related compounds, are probes for genetic variation in bitter taste, although PROP is safer with less sulfurous odor. Threshold for PROP distinguishes nontasters (increased threshold) from tasters (lower threshold); perceived intensity subdivides tasters into medium tasters (PROP is bitter) and supertasters (PROP is very bitter). Compared with supertasters, nontasters have fewer taste papillae on the anterior tongue (fungiform papillae) and experience less negative (e.g., bitterness) and more positive (eg, sweetness) sensations from alcohol. We determined whether the TAS2R38 gene at 7q36 predicted PROP bitterness, alcohol sensation and use. METHODS: Healthy adults (53 women, 31 men; mean age 36 years)--primarily light and moderate drinkers--reported the bitterness of five PROP concentrations (0.032-3.2 mM) and intensity of 50% ethanol on the general Labeled Magnitude Scale. PROP threshold and density of fungiform papillae were also measured. Subjects had common TAS2R38 gene haplotypes [alanine-valine-isoleucine (AVI) and proline-alanine-valine (PAV)]. RESULTS: PROP bitterness varied significantly across genotypes with repeated measures ANOVA: 26 AVI/AVI homozygotes tasted less bitterness than either 37 PAV/AVI heterozygotes or 21 PAV/PAV homozygotes. The PAV/PAV group exceeded the PAV/AVI group for bitterness only for the top PROP concentrations. The elevated bitterness was musch less than if we defined the groups using psychophysical criteria. With multiple regression analyses, greater bitterness from 3.2 mM PROP was a significant predictor of greater ethanol intensity and less alcohol intake- effects separate from age and sex. Genotype was a significant predictor of alcohol intake, but not ethanol intensity. With ANOVA, AVI/AVI homozygotes reported higher alcohol use than either PAV/AVI heterozygotes or PAV/PAV homozygotes. When age effects were minimized, PROP bitterness explained more variance in alcohol intake than did the TAS2R38 genotype. CONCLUSIONS: These results support taste genetic effects on alcohol intake. PROP bitterness serves as a marker of these effects. PMID- 15547450 TI - Alcohol increases glucagon secretion in normal man. PMID- 15547451 TI - Octanol modulation of neuronal nicotinic acetylcholine receptor single channels. AB - BACKGROUND: We have previously shown that alcohols exert a dual action on neuronal nicotinic acetylcholine receptors (AChRs), with short-chain alcohols potentiating and long-chain alcohols inhibiting acetylcholine (ACh)-induced whole cell currents. At the single-channel level, ethanol increased the channel open probability and prolonged the channel open time and burst duration. In this study, we examined the detailed mechanism of the inhibitory action of the long chain alcohol n-octanol on the neuronal nicotinic AChR. METHODS: Single-channel currents induced by application of 30 nm ACh were recorded with the patch-clamp technique from human embryonic kidney cells stably expressing the human alpha4beta2 AChR. RESULTS: Several single-channel parameters were markedly changed by octanol. At least two conductance-state currents were induced by low concentrations of ACh, and octanol increased the proportion of the low conductance-state current relative to the high-conductance-state current without changing the current amplitude. Major analyses of temporal properties of single channel currents were performed on the high-conductance-state currents. Octanol decreased the burst duration and duration of openings within burst and prolonged the mean closed time. All of these changes contributed to the decrease in the open probability in a concentration-dependent manner. CONCLUSIONS: Several aspects of octanol action on neuronal AChRs at the single-channel level are compatible with an atypical open channel block model reported with muscle nicotinic AChRs. The potentiating action of short-chain alcohols and the inhibitory action of long-chain alcohols on the neuronal nicotinic AChR are mediated through different mechanisms. PMID- 15547452 TI - Perceptual learning in detoxified alcoholic men: contributions from explicit memory, executive function, and age. AB - BACKGROUND: Visuospatial and visuoperceptual deficits have consistently been observed in detoxified alcoholics; however, the severity of impairment varies with test and task type. Identifying the component processes and factors that underlie a particular deficit may reveal why some visuospatial and visuoperceptual tasks are more compromised than others and may lead to the specification of neural systems that are particularly vulnerable in alcoholism. METHODS: We examined visuoperception and perceptual learning with a picture fragment identification task in 51 recently detoxified nonamnesic alcoholic men (aged 29-66 years) compared with 63 normal control men (aged 21-70 years). Executive function and explicit declarative memory were also assessed. RESULTS: Despite deficits in the primary components of visuoperception and explicit memory for visuospatial stimuli, the alcoholics showed normal perceptual learning. Although the alcoholics and controls performed at comparable levels on the perceptual learning task, multiple regression analyses indicated that the factors accounting for perceptual learning variance differed between and within groups. Visuoperceptual abilities consistently predicted perceptual learning in the control subjects but not the alcoholic subjects. Explicit memory contributed to perceptual learning performance in both the alcoholic and control groups. Frontal executive ability consistently predicted perceptual learning in the alcoholic subjects, but it had predictive ability only in the control subjects as time elapsed. Age was significantly correlated with perceptual learning performance in both groups. Lifetime alcohol consumption, but not alcoholism duration, was an independent predictor of 1-hr perceptual learning. CONCLUSIONS: These correlational analyses suggest that controls invoke basic visuospatial processes to perform a perceptual learning task, whereas alcoholics invoke higher-order cognitive processes (i.e., frontal executive systems) to perform the same task at normal levels. Use of more demanding cognitive systems by the alcoholics may be less efficient and more costly to processing capacity than those invoked by controls. PMID- 15547453 TI - The dopamine D1 antagonist reduces ethanol reward for C57BL/6 mice. AB - BACKGROUND: Dopamine D1 antagonist effects on behaviors related to obtaining and consuming ethanol remain unclear. The highly selective D1 antagonist ecopipam (SCH 39166), which has no effect on the serotonin system, was used to evaluate the role of D1 receptors in ethanol reward and its potential for treating alcohol abuse by determining its effect on several measures of ethanol reward in C57BL/6 (B6) mice. METHODS: Ecopipam (0.025-0.2 mg/kg) effects on instrumental and contingent consummatory responses and on noncontingent consummatory responses for ethanol and water reward were determined in food-restricted male mice trained to lever-respond for 12% ethanol delivered on a fixed ratio-4 reinforcement schedule. The mice were tested for 15-min sessions under preprandial (high-hunger and low-thirst) and postprandial (low-hunger and high-thirst) test conditions. RESULTS: Ecopipam dose-dependently reduced instrumental and consummatory responses for ethanol and ethanol intake when tested under hunger- or thirst motivated conditions with free access to water. Under thirst motivation with no access to an alternate fluid source, lever responses for ethanol and water were similar; however, ecopipam reduced responding for ethanol more than responding for water reward. When given concurrent free access to the same fluid delivered for lever pressing, animals made more contacts for ethanol than for water; ecopipam reduced free ethanol but not water contacts. CONCLUSIONS: Ecopipam attenuated ethanol reward at doses that did not affect water reward, indicating an effect independent of reductions in motor system function or general motivation and arousal. Ecopipam also reduced ethanol reward to the same degree under hunger, thirst, or sated conditions, again indicating that it affected ethanol reward at doses that did not grossly affect general motivational states. These data suggest that ecopipam may reduce ethanol reward with few side effects and that it warrants further investigation as a pharmacological tool for treating alcohol abuse. PMID- 15547454 TI - Enhanced alcohol self-administration after intermittent versus continuous alcohol vapor exposure. AB - BACKGROUND: Ethanol self-administering rats exhibit enhanced responding during withdrawal from continuous exposure to ethanol vapor. This study compared self administration of ethanol during withdrawal from continuous versus intermittent ethanol vapor. METHODS: Experiment 1 examined self-administration of ethanol in rats trained to self-administer ethanol after continuous, intermittent (14 hr on and 10 hr off), or no (i.e., controls) ethanol vapor exposure. Exposure time was equalized such that the intermittent group received 4 weeks of exposure and the continuous group received 2 weeks of exposure. Four self-administration tests were conducted 2 hr after removal from vapor, and each test was separated by 3 to 4 days of ethanol vapor. Experiment 2 examined self-administration of ethanol after 2 weeks of intermittent vapor either 2 or 8 hr after removal from vapor. Experiment 3 addressed the specificity of the increased responding for ethanol by examining saccharin self-administration after 2 weeks of intermittent vapor. RESULTS: Four weeks of intermittent exposure produced an increase in ethanol self administration during the first withdrawal relative to controls and relative to animals receiving 2 weeks of continuous exposure. The continuous group was indistinguishable from controls on the first test and gradually increased their responding across tests. Two weeks of intermittent exposure also increased ethanol self-administration, and there was no difference in this effect 2 or 8 hr after removal from vapor. There was no difference in saccharin self administration in control rats and those given 2 weeks of intermittent exposure. CONCLUSIONS: The finding that intermittent exposure produces more rapid increases in self-administration of ethanol relative to continuous exposure suggests that intermittent exposure may be associated with a more rapid escalation of the allostatic processes responsible for excessive ethanol self-administration. The mechanisms that drive the increases in drinking during withdrawal are similar after 2 and 8 hr of withdrawal and seem to be specific to ethanol. PMID- 15547455 TI - Blockade of the leptin-sensitive pathway markedly reduces alcohol consumption in mice. AB - BACKGROUND: The neuropeptide leptin links adipose stores with hypothalamic centers and serves as an endocrine signal involved in the regulation of appetite (and possibly in the endorphinergic modulation of the drug reward system). Increased plasma leptin has been observed at the onset of alcohol withdrawal in humans, and ethanol consumption after withdrawal was increased by injection of leptin in mice. We addressed the role of leptin in alcohol-related behaviors by studying ethanol consumption in two strains of spontaneously mutant mice that lack leptin (ob/ob) or the leptin receptor (db/db). METHODS: Two strains of mutant leptin-deficient (ob/ob) or leptin-resistant (db/db) mice were tested in a two-bottle-choice paradigm and were compared with wild-type (C57BL/6 inbred strain) mice. The effects of leptin injection on voluntary ethanol intake have been investigated in ob/ob and C57BL/6 mice. RESULTS: Males and females of both mutant strains showed a significantly lower preference for alcohol in a two bottle-choice paradigm compared with wild-type mice. Male ob/ob mice demonstrated slightly higher avoidance of bitter taste, and females of the both mutant strains showed a reduced preference for saccharin solutions. Administration of leptin (1 mg/kg intraperitoneally, daily for 8 days) altered body weight but failed to increase the preference for ethanol in ob/ob mice; i.e., we could not correct the effects of leptin deficiency on alcohol consumption by the injection of leptin. Also, there were no differences between the effects of leptin (1 mg/kg intraperitoneally, daily for 8 days) and saline injections on alcohol consumption in C57BL/6 mice. CONCLUSIONS: These data show that blockade of the leptin pathway markedly decreases the preference for alcohol intake, but this decrease may be the result of compensatory or developmental changes in other systems rather than a more direct effect of leptin on alcohol consumption. PMID- 15547456 TI - Gender differences in the performance of a computerized version of the alcohol use disorders identification test in subcritically injured patients who are admitted to the emergency department. AB - OBJECTIVE: The Alcohol Use Disorder Identification Test (AUDIT) has been recommended as a screening tool to detect patients who are appropriate candidates for brief, preventive alcohol interventions. Lower AUDIT cutoff scores have been proposed for women; however, the appropriate value remains unknown. The primary purpose of this study was to determine the optimal AUDIT cutpoint for detecting alcohol problems in subcritically injured male and female patients who are treated in the emergency department (ED). An additional purpose of the study was to determine whether computerized screening for alcohol problems is feasible in this setting. METHODS: The study was performed in the ED of a large, urban university teaching hospital. During an 8-month period, 1205 male and 722 female injured patients were screened using an interactive computerized lifestyle assessment that included the AUDIT as an embedded component. World Health Organization criteria were used to define alcohol dependence and harmful drinking. World Health Organization criteria for excessive consumption were used to define high-risk drinking. The ability of the AUDIT to classify appropriately male and female patients as having one of these three conditions was the primary outcome measure. RESULTS: Criteria for any alcohol use disorder were present in 17.5% of men and 6.8% of women. The overall accuracy of the AUDIT was good to excellent. At a specificity >0.80, sensitivity was 0.75 for men using a cutoff of 8 points and 0.84 for women using a cutoff of 5 points. Eighty-five percent of patients completed computerized screening without the need for additional help. CONCLUSIONS: Different AUDIT scoring thresholds for men and women are required to achieve comparable sensitivity and specificity when using the AUDIT to screen injured patients in the ED. Computerized AUDIT administration is feasible and may help to overcome time limitations that may compromise screening in this busy clinical environment. PMID- 15547457 TI - Magnesium treatment of primary alcohol-dependent patients during subacute withdrawal: an open pilot study with polysomnography. AB - BACKGROUND: Sleep electroencephalogram alterations and insomnia complaints persist after alcohol withdrawal in dependent patients and are considered strong predictors of relapse. Although disturbances of magnesium household due to alcohol consumption are well known, the relationship of magnesium metabolism and sleep disturbances has not been investigated in this patient group. We conducted an open pilot study to evaluate the effects of magnesium treatment on the sleep of primary alcohol-dependent patients during subacute withdrawal. METHODS: Patients were treated with 30 mmol magnesium daily over 4 weeks. Eleven of the 14 included patients were evaluated. Patients were free of any kind of psychotropic medication or other substances known to influence sleep. Polysomnographic recordings with monitoring of periodic leg movements in sleep (PLMS) were performed for two consecutive nights 2 weeks after acute withdrawal (baseline) and 4 weeks later at the end of the treatment period. After the baseline polysomnography, patients were investigated by the magnesium loading test to verify magnesium depletion. RESULTS: We found a significant decrease of sleep onset latency from 40.6 to 21.7 min (p = 0.03) and a significant improvement of subjective sleep quality, as assessed by the Pittsburgh Sleep Quality Index, from 8.1 to 5.8 (p = 0.05) during magnesium treatment. Changes in PLMS indices revealed two subgroups of patients: one with an increase of PLMS from 30.7 to 39.4 per hour of sleep (n = 4) and the other one with a decrease of PLMS from 8.9 to 2.1 per hour of sleep (p = 0.04). Patients with PLMS decreases seemed to have a more favorable prognosis: total sleep time, gamma-glutamyltransferase, carbohydrate-deficient transferrin, and Beck Depression Inventory scores improved significantly during treatment in this group. The magnesium loading test revealed a magnesium deficiency in only one patient, five patients showed normal retention values, and the remaining five patients had an increased magnesium excretion, indicating a possible continued renal magnesium loss during abstinence. CONCLUSIONS: The results of this study should be interpreted with caution, because no control group with placebo was investigated. Both subjective and, partly, objective parameters of sleep improved during the 4-week study period. Further research is needed to clarify the relationship of magnesium metabolism and sleep alterations. PMID- 15547458 TI - Effectiveness of naltrexone in a community treatment program. AB - OBJECTIVE: In several large, well-designed, randomized, double-blind studies, the opiate antagonist naltrexone demonstrated efficacy in the treatment of alcohol dependence. Specifically, when combined with certain psychosocial therapies, naltrexone reduces the number of drinking days, heavy drinking, and time to relapse to alcohol use in alcohol-dependent individuals. Whether this efficacy can be generalized to individuals who have alcohol use disorders and present for treatment at front-line community treatment programs has not been well established. METHODS: A total of 145 patients who presented for treatment at a rural community substance abuse treatment center were randomized to receive naltrexone 50 mg daily plus usual program treatment (n = 54), placebo plus usual treatment (n = 43), or usual treatment alone (n = 48) for 12 week. A total of 133 participants had at least one follow-up visit. Primary outcome measures included percent days drinking, average drinks per drinking day, average drinks per day, heavy drinking days (four or more for women and six or more for men), and time to first heavy drinking day. Secondary measures included changes in serum biological markers (alkaline phosphatase, alanine transaminase, aspartate transaminase, and gamma-glutamyltransferase), craving, and psychosocial functioning. RESULTS: In the intention-to-treat analysis, there were no between-group differences for any of the primary drinking outcomes at 12 weeks. In post hoc exploratory analyses, the entire sample of participants was divided into two new groups: (1) people who drank during the 2 weeks before the start of medication (entry drinkers) and (2) people who did not drink during this interval (entry abstainers). Entry abstainers were at an advantage at study entry in that they were significantly more likely to have an inpatient hospitalization immediately before entry into outpatient treatment. Mixed-model analysis of variance revealed a main effect for entry group at the 12-week treatment endpoint on the primary outcome measures of percent days drinking, average drinks per drinking day, average drinks per day, heavy drinking days, and time to first heavy drinking day. Participants in any of the randomized groups who were entry abstainers had significantly better improvement on all of the primary outcome measures. The abstainer groups that were randomized to placebo and usual treatment had significantly better outcomes than the entry drinkers in those perspective groups. However, for the naltrexone treated group, entry drinkers and entry abstainers had similar improvement in drinking-related outcomes. CONCLUSIONS: These data suggest that naltrexone may offer particular benefit to patients who continue to drink during the early stages of the trial as compared with those who have achieved abstinence before treatment entry. PMID- 15547459 TI - Phosphatidylethanol in human organs and blood: a study on autopsy material and influences by storage conditions. AB - OBJECTIVE: Phosphatidylethanol (PEth) is an abnormal phospholipid that is formed and accumulated in mammalian cells that have been exposed to ethanol. PEth has been proposed as a marker of ethanol abuse. This study was conducted to investigate the concentration of PEth in blood and organs obtained during the autopsy of alcoholics. In addition, we performed experiments on rat tissues and human blood to evaluate the effect of various storage conditions on PEth concentrations. METHODS: Human tissues and blood from alcoholics and controls were obtained at autopsy and frozen at -20 degrees C until extraction. Blood from healthy donors was incubated with ethanol for 24 hr and thereafter either extracted directly or stored at room temperature, stored at 4 degrees C, frozen at -20 degrees C, or frozen in liquid nitrogen and stored at -80 degrees C before extraction. Rats were given intraperitoneal injections of ethanol and then killed, either while still intoxicated or when sober. Rat organs were homogenized and extracted directly, after a period of storage, and/or after freezing at -20 degrees C. PEth concentration was analyzed using HPLC and verified by mass spectrometry. RESULTS: In all rat organs studied, PEth was formed during freezing at -20 degrees C with ethanol present. PEth concentrations of 9 to 205 mumol/liter were observed in the blood obtained at autopsy. The highest value was found in the case with the highest blood alcohol concentration (114 mmol/liter) at the time of death. In the experiments on human blood stored with ethanol present, PEth concentrations were not affected after 72 hr at 4 degrees C or after freezing in liquid nitrogen and storage at -80 degrees C for up to 144 hr but were slightly elevated after 24 hr at room temperature and at -20 degrees C. PEth was found in all organs obtained from the cadavers of alcoholics. Storage of organs at 4 degrees C for 24 hr with ethanol present had no effect on the PEth concentration. The PEth concentration was unaffected when no ethanol was present at the time of freezing. CONCLUSIONS: The rat experiments indicated that the very high PEth concentrations found in the organs of the alcoholics were probably largely formed while the organs were frozen at -20 degrees C. Our data suggest that tissue material from bodies that were exposed to ethanol must be stored properly to obtain reliable results from subsequent analysis for PEth. Tissue should not be frozen at -20 degrees C but instead stored refrigerated until extraction, preferably within hours of autopsy, or frozen in liquid nitrogen and stored at -80 degrees C. Blood samples that contain ethanol can be stored refrigerated for up to 72 hr or frozen in liquid nitrogen and stored at -80 degrees C without affecting PEth levels. PMID- 15547460 TI - Clinical practice characteristics and preconception counseling strategies of health care providers who recommend alcohol abstinence during pregnancy. AB - OBJECTIVE: National initiatives on fetal alcohol syndrome in Canada and the United States aimed at prevention, identification, and treatment of individuals who are affected by alcohol exposure in utero recommend that women abstain from consuming alcohol during pregnancy. Health care providers are key educators regarding appropriate alcohol use. The objective of this study was to describe characteristics of physicians who recommend alcohol abstinence during pregnancy with regard to knowledge of fetal alcohol syndrome and preconception counseling strategies. METHODS: A survey was mailed to Canadian physicians and midwives between 2001 and 2002. Participants consisted of a national random sample of 1090 Canadian obstetricians and gynecologists, midwives, and family physicians who were current members of provincial and national professional organizations. The main outcome measure was questionnaire responses to knowledge, prevention, and diagnosis of issues related to alcohol use during pregnancy. RESULTS: Response rates ranged from 31.1% among family physicians to 63.5% among midwives. Overall, 91.2% of providers recommended abstinence from alcohol during pregnancy. These providers were significantly more likely to believe that there is sufficient information about alcohol use and that clients were interested in discussing alcohol (p < 0.05). They were also significantly more likely to discuss depression, personal alcohol use, partner's use of alcohol, and family history of alcohol misuse with women of childbearing age (p < 0.05). Once a patient became pregnant, fewer practice differences were noted, although those who recommended alcohol abstinence were significantly more likely to take clinical action when pregnant patients were consuming moderate amounts of alcohol (p < 0.05). CONCLUSIONS: It is encouraging that almost 90% of Canadian health care providers recommend abstinence from alcohol during pregnancy. However, differences in clinical practice exist between providers who recommend alcohol abstinence during pregnancy as compared with those who recommend a "glass in moderation." PMID- 15547461 TI - Maternal age, alcohol abuse history, and quality of parenting as moderators of the effects of prenatal alcohol exposure on 7.5-year intellectual function. AB - BACKGROUND: In contrast to the extensive literature documenting IQ deficits in patients with fetal alcohol syndrome, effects on IQ have not generally been reported for children with alcohol-related neurodevelopmental disorder (ARND). This study examined the role of maternal age, MAST, and quality of parenting in moderating the effects of prenatal alcohol exposure on the WISC-III IQ test in moderate-to-heavily exposed children. METHODS: A total of 337 inner-city African American children whose mothers were recruited prenatally were administered the WISC-III at 7.5 years. Alcohol exposure was assesed with a timeline follow-back interview administered at every prenatal clinic visit. Moderating effects of the three risk factors were examined by adding interaction terms to regression analyses and dichotomizing the moderators and performing separate regressions on the two groups. RESULTS: Prenatal alcohol exposure was related to WISC-III Freedom from Distractibility but not to Full Scale IQ for the sample as a whole. However, among children born to older mothers, an alcohol effect emerged on Full Scale IQ and five of seven composite IQ scores. Similarly, adverse effects were seen among children of MAST-positive mothers and children whose parents provided less optimal cognitive stimulation. Each additional ounce of absolute alcohol consumed per day during pregnancy was associated with a 2.9-point decrease in Full Scale IQ and a 5.6-point decrement on Freedom from Distractibility. CONCLUSIONS: This study is the first to demonstrate IQ effects among children with ARND born to older and MAST-positive mothers, particularly in relation to first-trimester drinking. These findings suggest that there are subgroups of more vulnerable and severely affected children whose intellectual performance is compromised. A moderate- to heavy-drinking mother who has given birth to an unaffected child when she was younger needs to be warned that her risk of having an adversely affected child increases as she grows older. PMID- 15547462 TI - Inhibition of nitric oxide release in vivo by ethanol. AB - OBJECTIVE: Previous studies indicate that the nitric oxide (NO(.)) pathway is involved in the acute or chronic effects of ethanol on the central nervous system. However, direct evidence for the effect of ethanol on NO(.) production in vivo is lacking, and it is not clear whether it is inhibition or stimulation of the NO(.) pathway that contributes to the behavioral effects of ethanol. Herein the release of NO(.) in the rat striatum in vivo in response to NMDA receptor activation--the dominant mechanism controlling NO(.) formation-has been investigated after systemic or local injections of ethanol. METHODS: NMDA-induced release of authentic NO(.) was measured directly in the striatum of urethane anesthetized (1.2 g/kg intraperitoneally) male Sprague-Dawley rats by using a direct-current amperometric method coupled to an electrically modified carbon microelectrode. An injector cannula was implanted in the proximity of the electrode (250 microm apart) for focal drugs application. RESULTS: Local application of NMDA (1 microl, 100 microM) produced a sharp and transient NO(.) signal. Systemic ethanol, 1 or 2.5 g/kg intraperitoneally, caused a long-lasting, dose-dependent inhibition of NMDA-induced NO(.) release to 12.2 +/- 5.9 and 6.4 +/- 3.7% of control, respectively, 60 min after ethanol administration. Dizocilpine (0.5 mg/kg intraperitoneally) mimicked the ethanol effect, inhibiting NO release to 1.6 +/- 0.66% of control. Local application of ethanol (1 microl, 2.5% v/v) in the striatum reduced the NMDA-induced response to 28.6 +/- 3.8% of control. Focal application of the competitive NMDA receptor antagonist D-(-)-2 amino-5-phosphonopentanoic acid (100 microM) or the nonselective NO synthase inhibitor L-N(G)-nitro-arginine methyl esther (100 microM) also caused inhibition of NMDA-induced NO(.) release to 2.4 +/- 0.7 and 4.3 +/- 0.9% of control, respectively. CONCLUSIONS: Ethanol, at pharmacologically significant doses, strongly inhibits striatal NO(.) production and release apparently through inhibition of NMDA receptor function. Inhibition of NMDA receptor-mediated activation of the NO(.) pathway could be a primary neurobiological mechanism contributing to the effects of ethanol. PMID- 15547463 TI - Ethanol induces expression of the glutamate transporters EAAT1 and EAAT2 in organotypic cortical slice cultures. AB - BACKGROUND: Exposure of the developing brain to ethanol disposes the fetus to fetal alcohol syndrome and causes a number of changes in several neurochemical systems. In particular, the glutamatergic system is affected by ethanol. Thus, increased sensitivity of glutamate receptors and enhanced transmembrane transport of glutamate were found in primary astrocyte cultures. However, in these in vitro studies, changes in the expression of glutamate transporters were not detected. To further characterize the influence of chronic ethanol exposure on the developing brain, we assessed the transcriptional and translational regulation of glutamate transporters in a less artificial in vitro system. METHODS: We exposed postnatal rat cortical organotypic slice cultures to ethanol (100 mM) for 4 and 8 days. Expression of the excitatory amino acid transporters EAAT1 and EAAT2 was analyzed in comparison to untreated controls with semiquantitative reverse transcriptase-polymerase chain reaction. In addition, changes in protein expression were detected by Western blotting. RESULTS: In ethanol-exposed cortical slices, we observed more prominent fiber outgrowth and significantly increased gene expression (EAAT1: +35%, p = 0.029; EAAT2: +251%, p = 0.015). These findings could be verified on the protein level, because Western blot analysis revealed significantly higher levels of EAAT1 (+76%; p = 0.008) and EAAT2 (+104%; p = 0.018) in ethanol-treated slices compared with controls. CONCLUSIONS: Our results are in concert with earlier studies describing the induction of glutamate transport by ethanol. Enhanced expression of EAAT1 and EAAT2 after ethanol exposure can be viewed as a maladaptive process that disposes the developing brain to fetal alcohol syndrome. PMID- 15547464 TI - Alcohol intoxication impairs phosphorylation of S6K1 and S6 in skeletal muscle independently of ethanol metabolism. AB - BACKGROUND: The purpose of this study was to characterize the ability of alcohol to suppress insulin-like growth factor (IGF)-I stimulation of ribosomal S6 kinase 1 (S6K1) and 4E-BP1 phosphorylation, which are central elements in the signal transduction pathway used to coordinate the protein synthetic response and may contribute to the development of alcoholic myopathy. METHODS: In vivo studies examined the dose and time dependency of the ability of alcohol to impair signal transduction under basal and IGF-I-stimulated conditions. Additional studies examined the effect of gender, nutritional state, and route of alcohol administration. A separate study determined the direct effects of alcohol on muscle metabolism by using the isolated perfused hindlimb preparation. RESULTS: The phosphorylation of S6K1 and S6 in muscle was increased after injection of IGF I in control rats. In contrast, IGF-I failed to stimulate S6K1 or S6 phosphorylation 2.5 hr after intraperitoneal administration of alcohol when the blood alcohol concentration was increased between approximately 165 and 300 mg/dl. With a maximal suppressive dose of alcohol, the inhibitory effect on S6K1/S6 phosphorylation was observed as early as 1 hr and for up to 8 hr. The ability of alcohol to impair phosphorylation of S6K1 and S6 was independent of gender (male versus female), nutritional status (fed versus fasted), and route of alcohol administration (intraperitoneal versus oral). Furthermore, the suppressive effect of alcohol was still observed in rats pretreated with 4 methylpyrazole, suggesting that the response was independent of the oxidative metabolism of ethanol. The direct effect of alcohol on IGF-stimulated S6K1/S6 phosphorylation was also present when the isolated hindlimb was perfused in situ with buffer containing alcohol. In contrast to S6K1, acute alcohol intoxication did not consistently impair the ability of IGF-I to stimulate 4E-BP1 phosphorylation under any of the experimental conditions. CONCLUSIONS: These data indicate that acute alcohol intoxication selectively impairs IGF-I signaling via S6K1, but not 4E-BP1, and that this defect is independent of gender, nutritional state, route of administration, and alcohol metabolism. The IGF-I resistance may represent a participating mechanism by which alcohol directly limits the translation of selected messenger RNAs and, ultimately, protein synthesis in skeletal muscle. PMID- 15547465 TI - [Anterior segment of the eye and diabetes mellitus]. AB - OBJECTIVE: To study the damage to the anterior segment of the eye during mellitus diabetes. METHODS: We prospectively studied the anterior segment of the eye in 200 diabetic patients and 200 patients with no history of diabetes (control group). Parameters included the corneal curvature radius, corneal thickness, central corneal sensitivity (Cochet Bonnet esthesiometer), pupil diameter before and after dilation using tropicamide and phenylephrine eye drops, delay for maximal pupil dilation, lens transparency, and intraocular pressure. The damage to the anterior segment was compared in both groups and was correlated with the parameters of diabetes (type, duration, and equilibration), retinopathy grade, and peripheral neuropathy. RESULTS: Corneal geometry (i.e., curvature radius and thickness) and intraocular pressure did not correlate with diabetes. Cataract was significantly influenced by diabetes (p<0.001). The duration needed to obtain pharmacological maximal mydriasis significantly correlated with iris color (rs=0.47, p<0.001), patient age (rs=0.28, p<0.001) and diabetes (rs=0.25, p<0.001). Dilation was significantly longer in the diabetic patient than in those of the control group (36.5 min vs 30.7 min, p<0.001). Corneal sensitivity significantly correlated with age (rs=0.48, p<0.001) and diabetes (rs=0.53, p<0.001). Corneal sensitivity was decreased in the diabetic group (5.42 vs 5.94, p<0.001) and it correlated with retinopathy grade (rs=0.47, p<0.001) and with peripheral neuropathy (rs=0.15, p<0.001). Sensitivity and specificity of hypoesthesia in detecting peripheral neuropathy were 67.4% and 49.4%, respectively. Among the parameters of diabetes, duration of progression showed the strongest correlation with damage to anterior segment (rs > or = 0.25, p<0.001). CONCLUSION: Diabetes influences lens transparency, pharmacological pupil dilatation and corneal sensitivity. In spite of the unspecific character of the damage and the subjectivity of the measurement, corneal sensitivity is an interesting parameter to take into consideration among diabetic patients. PMID- 15547466 TI - [Calcification of hydrophilic acrylic intraocular lenses: clinical and pathological aspects]. AB - PURPOSE: The aim of this study was to evaluate the clinical aspects of ten eyes with calcified hydrophilic acrylic intraocular lenses and pathological data obtained from seven explanted lenses. MATERIAL AND METHODS: Forty-seven eyes of 40 patients received the same implant in the first 6-month period of 2001. Ten eyes showed intraocular lens opacification detected 6-18 months after the operation: seven lenses were explanted and three were left in place because they were not causing a decrease in visual acuity or glare at light. Five of ten eyes were diabetic. The explanted lenses were examined under the light microscope and the electron microscope. The elemental analysis of the lens surfaces was made by energy dispersive spectrometry. RESULTS: The light microscopy showed an irregular surface covered by a gray-white opacity. The electron microscopy detected multiple granulations on the front and back surfaces of the lenses including some portions of the haptics. The size and density of these granulations were smaller on the back surface. The energy dispersive spectrometry showed the presence of calcium and phosphate on both surfaces. The spikes of calcium and phosphate were smaller for the back surface of the lenses. DISCUSSION: Calcification was predominantly seen on the surfaces that were in contact with aqueous not covered with anterior capsule. Half (5/10) of the cases were diabetic even though 18% of all patients receiving this lens were diabetic. The presence of diabetes is very common in other series. These data suggest the role of a metabolic factor influencing the milieu of the lens in this calcification process. CONCLUSION: Calcification of the hydrophilic acrylic lenses is a relatively serious complication, but the conditions leading to its appearance and the physiopathology have not yet been fully elucidated. The surgeon should be very careful in the choice of the intraocular lens to implant, and even more so if the patient is diabetic. PMID- 15547467 TI - [Bringing back scleral contact lenses]. AB - PURPOSE: Although old generations of scleral contact lenses were totally replaced by soft and hard contact lenses, new designs and materials have recently allowed the successful use of nonfenestrated scleral contact lenses in patients who are intolerant to regular contact lenses. These new scleral contact lenses form a bridge over the cornea, which is thus permanently bathed in a saline reservoir. METHODS: Retrospective study in patients suffering from corneal diseases altering the vision and/or responsible for ocular discomfort, and who are intolerant to conventional contact lenses. RESULTS: Fifteen eyes (10 patients) were fitted with specially designed scleral contact lenses. The mean follow-up was 16 months (range, 1-31 months). The scleral lenses were indicated for optical and/or therapeutic reasons: keratoconus (3 eyes), corneal grafts (7 eyes), refractive surgery (2 eyes), corneal dysplasia (2 eyes), and Stevens-Johnson syndrome (1 eye). Optical results and tolerance were excellent except for one eye. CONCLUSION: New nonfenestrated scleral contact lenses provide a good alternative for patients who are intolerant to conventional contact lenses. PMID- 15547468 TI - [Managing of retinopathy of prematurity in a tertiary center]. AB - INTRODUCTION: Retinopathy of prematurity is a complication of prematurity whose risk factors are well known. Its prognosis remains fearsome and its treatment is difficult. PATIENTS AND METHODS: Fifty-four children were managed (108 eyes analyzed) for retinopathy of prematurity in the Ophthalmology Ward of Necker Enfants-Malades Hospital between 1996 and 2002. A retrospective review of those cases was performed to describe their clinical features, their treatment and their outcome. RESULTS: Mean gestational age was 26.75 weeks after the mother's last period (24-32.5 weeks) and mean birth weight was 842 g (530-2260 g). Stage zero occurred in ten eyes, stage 1 in two eyes, stage 2 in seven eyes, stage 3 in 42 eyes, stage 4 in ten eyes, stage 5 in 24 eyes, and after-effects stage in 13 eyes. Initial treatment (combined treatments possible) was abstention in 45 eyes, cryotherapy in 16 eyes, laser photocoagulation in 43 eyes, surgical treatment of 13 eyes (scleral buckling or vitrectomy). Cryotherapy was effective in 71.5% of cases, photocoagulation was effective in 77%, and scleral buckling was effective in 25%. Vitrectomy failed in the five eyes treated. Outcome was complicated by legal blindness in 33 eyes. DISCUSSION AND CONCLUSION: The cases of retinopathy of prematurity that we manage are often severe, stage 3 or above, which explains the outcome of the patients. The 7-year experience reported herein shows how difficult it is to manage this condition, though the need for its detection, diagnosis criteria, and therapeutical indications are well known. Because younger and smaller premature newborns are successfully managed by pediatricians, retinopathy of prematurity is far from having vanished and its morbidity is considerable. PMID- 15547469 TI - [Clinical advantages of colorimetric exploration in the early diagnosis of glaucomatous neuropathy]. AB - AIM: The purpose of this investigation was to evaluate acquired color vision defects in the early stages of primary glaucoma and in patients at risk for glaucoma. MATERIAL AND METHODS: Patients with primary open-angle glaucoma, normal tension glaucoma and ocular hypertension were examined with the IF-2 anomaloscope. The Rayleigh equation and the Moreland equation were tested. We studied the relationships between color vision defects and the other tested variables. RESULTS: We found the mean value of the setting range of the Moreland equation significantly increased in all tested groups. Color vision disturbances were found in 57% of eyes with glaucoma and in 31% of eyes with ocular hypertension. We found a positive correlation between the blue-green colour vision defects and the perimetric changes in the glaucomatous eyes. These results indicate a decrease in color discriminating sensitivity in the short wavelength part of the visible spectrum in patients with glaucoma as well as in patients with ocular hypertension. CONCLUSION: Blue-yellow color vision testing with the anomaloscope may serve as an additional test in the early diagnosis of glaucoma but the absence of dyschromatopsia does not exclude the disease. PMID- 15547470 TI - [The Mydriasert insert: an alternative to eye drops for preangiographic mydriasis]. AB - PURPOSE: Mydriasert is an insoluble ophthalmic insert, gradually releasing two well-known active ingredients: phenylephrine and tropicamide. It is indicated in presurgical mydriasis. The purpose was to evaluate its efficacy in obtaining mydriasis required for fluorescein angiography. MATERIAL AND METHODS: The ability of Mydriasert to provide mydriasis (defined by a pupillary diameter of at least 7 mm) compatible with a bilateral angiographic examination in optimal conditions was evaluated by a comparative, randomised versus active treatment (eye drops) open trial in 72 patients. RESULTS: All the patients obtained a stable mydriasis allowing angiography. In the Mydriasert group, mydriasis preparation required a mean of 10 min more (Student t test: p<0.001); however, near eyesight recovery was 15 min shorter on average (log-rank test<0.01%) and amounts of active ingredients administered to provide mydriasis were 5-10 times higher in the eye drops reference group. Cardiovascular parameters remained in the normal range in both groups. Three patients of the Mydriasert group presented one ocular symptom of local intolerance, which disappeared in 15 min (exact Fisher test not significant between the treatment groups, p=0.2394). No superficial punctate keratitis was detected during the trial. CONCLUSION: The treatment by Mydriasert can prepare preangiographic mydriasis as well as the reference treatment. The time required for mydriasis is slightly longer. Near eyesight recovery, faster with Mydriasert, could provide an improvement in patient safety and comfort at the end of the ophthalmologic visit. PMID- 15547471 TI - [Modified evisceration for biocolonizable orbital implant: the four-square technique]. AB - INTRODUCTION: The introduction of a porous, biocolonizable, orbital implant is difficult after a classic evisceration. We have developed a modification of the method that provides better results. MATERIAL AND METHODS: Four scleral squares pediculized on the right oculomotor muscles were cut. They were sutured two-by two over the anterior part of the implant. RESULTS: Fifteen patients were operated. There were no complications during a mean follow-up of 12.4 months. DISCUSSION: The soft anterior tissues were protected by the two vascularized scleral layers. The diameter of the implant was 18 mm for three patients, 20 mm for eight patients, and 22 mm for four patients. Operating time was much shorter than with enucleation and covering of the implant with autologous sclera. CONCLUSION: Evisceration with the four-square technique is a safe and quick method, which can be used for most patients, except those with an intraocular neoplasia. PMID- 15547472 TI - [Nonpenetrating deep trabeculectomy treated with mitomycin C, without implant. A prospective evaluation of 55 cases]. AB - PURPOSE: Since 1998 we have been conducting a prospective study of nonpenetrating deep trabeculectomy with chronic open-angle glaucoma to evaluate the efficiency of the technique. MATERIAL AND METHODS: The study was carried out in 55 eyes of 41 patients who suffered from open-angle glaucoma. After performing a superior scleral flap, mitomycin diluted to 0.01% was applied for 3 minutes, then the 4 x 4-mm superficial scleral flap was dissected at two-thirds deepness until reaching the cornea. The Schlemm canal and the external trabecula were surgically removed and the two points of the Schlemm canal were catheterized with a trabeculotome to ensure that the ablation was well done. If it was not, it was completed by using a trabeculotome as a guide. Postoperatively, if the filtering bleb tended to decrease or ocular pressure began to increase, the operated trabecular region was reopened with Yag laser. The filtering bleb characteristics were correlated with the normalization of intraocular pressure in the first 30 cases. RESULTS: Preoperative pressure without treatment was 32 mmHg. Postoperative intraocular pressure without treatment was 20 mmHg or less in 79% of the eyes after 4 months, 77.5% after 6 months, 75% after 8 months and 61% after 12 months. By adding a local hypotension treatment in monotherapy, a pressure of 20 mmHg or less was obtained in 79% of the cases after 12 months. No severe complications were observed. The presence of a filtering bleb is an important factor in the normalization of postoperative pressure (p=0.0048). CONCLUSIONS: This surgical technique provides a substantial decrease in intraocular pressure and very few complications after 12 months of follow-up. PMID- 15547473 TI - [Teaching phacosection in a tropical setting]. AB - PURPOSE: The aim of this study was to prospectively evaluate the training of a Malagasy ophthalmologist, already proficient in classic manual extracapsular cataract extraction (ECCE), in a small incision manual technique (phacosection). MATERIALS AND METHODS: Within the activity of the Sight First program to fight blindness in Madagascar, the ophthalmologist of Majunga recruited patients presenting total white cataracts. These patients were operated by phacosection using the local hospital's usual instrumentation along with two specific phacosection instruments and extra single-use material (a precalibrated 3.2 mm knife and viscoelastic products). Peribulbar anesthesia with eye pressure was provided. Postoperative follow-up (visual acuity and corneal status) was conducted on days 1, 7 and 30. RESULTS: Forty-four eyes were operated, the first seven by the instructor assisted by the local ophthalmologist, the next 36 by the local ophthalmologist assisted by the instructor. Five postoperative complications were noted: two cases of posterior capsular rupture (one during implantation) and three cases needing reoperation with aspiration of residual cortical masses. Induced astigmatism was low and visual rehabilitation was good, with all corneas clear at day 30. DISCUSSION: The results are quite acceptable given the training period, with only two marked complications (one not related to the operative technique) and good visual rehabilitation in less than 2 weeks. This small incision technique allowed reduction of induced astigmatism and risk free management of total white cataracts (independent of operator). CONCLUSION: Small incision manual ECCE by phacosection is a safe, possible management option for difficult cataracts in a tropical setting with only a small increase in cost. Moreover, the training of an ophthalmologist mastering classic manual ECCE appears rapid and risk-free for patients. PMID- 15547474 TI - [Granulomatous uveitis and CREST syndrome: a case study]. AB - PURPOSE: To report a case of recurrent granulomatous panuveitis associated with CREST syndrome. OBSERVATION: A 74-year-old patient with CREST syndrome presented with unilateral granulomatous panuveitis in a pseudophakic eye. She had undergone cataract surgery 6 months before. The patient reported a vision loss that had been evolving for 1 month. Visual acuity was noted at 20/400. The initial clinical examination highlighted retrodescemetic precipitates and granulomatous precipitates on the IOL. A vitreous tyndall was noted. Funduscopic examination revealed papillary edema and cystoid macular edema, confirmed by fluorescein angiography. RESULTS: Topical treatment consisting in corticosteroid eye drops associated with mydriatics controlled uveitis in a few weeks. Visual recovery was 20/30. No granulomatous uveitis etiology could be highlighted. The diagnosis of chronic endophthalmitis was also ruled out. CONCLUSION: The diagnosis retained was uveitis associated with CREST syndrome. To our knowledge, this association has only been reported twice in the literature. PMID- 15547475 TI - [Optic nerve ischemia as a complication of dental extraction: a case report]. AB - We report the clinical observation of a 16-year-old male, with no particular medical or ocular history, who presented with loss of vision in the right eye on the first post-operative day after uncomplicated extraction of the four third molar roots under general anesthesia. Loss of vision, due to optic disc ischemia, persisted after 2 months. The clinical and pathogenic features of this rare optic nerve ischemia complication are discussed. PMID- 15547476 TI - [Eales disease and tuberculous allergy]. AB - The authors report the case of a 25-year-old woman who presented unilateral Eales disease associated with biologically confirmed pulmonary tuberculosis. The patient, from a family with a history of tuberculosis, showed a painless and abrupt decrease in visual acuity of the right eye with venous and arterial vasculitis. Fluorescein angiography confirmed the existence of peripheral ischemia and vascular abnormalities. The biological exam showed a positive PPD and the BK was isolated. Antibiotic and corticosteroid drugs were used in the treatment. The clinical, immunopathological and therapeutic aspects of this disease are discussed. PMID- 15547477 TI - [Purtscher retinopathy in acute alcoholic pancreatitis. A case report]. AB - Purtscher retinopathy is a retinal vasculopathy most commonly caused by head or chest trauma. The most frequent signs include white ischemic infarcts (cotton wool spots) and retinal hemorrhages (intraretinal, preretinal, or flame). The signs of ischemic vasculopathy predominate in the interpapillomacular zone. We report a case of a 31-year-old woman with sudden bilateral loss of vision caused by Purtscher retinopathy in acute alcoholic pancreatitis. PMID- 15547478 TI - [Hormonal regulatory influence in tear film]. AB - Tear film quality depends on fine regulatory mechanisms affected by neuronal and hormonal influences. Indeed, receptors for androgens, estrogens, progesterone and prolactin have been identified in several ocular tissues in the rat, rabbit and in humans. The eye is thus a target organ for sex hormones, particularly the androgens. These hormones regulate the immune system, the morphology and secretory functions of lacrimal glands and the functioning of Meibomian glands. The influence of hormone replacement therapy in menopausal women remains unclear, as some authors support the idea that they improve the quality and the volume of tear film, whereas others have shown that they increase the risk of dry eye. Finally, knowledge of the interactions between the hormones that influence the lacrimal glands is essential for the understanding of the regulation of lacrimal gland function. However, the data presently available strongly suggest that optimal bioavailable androgen levels are essential for normal lacrimal gland function and that prolactin and estrogens also play important roles in providing a hormonal milieu that contributes to normal lacrimal gland function. PMID- 15547480 TI - [Fundamentals of the anatomy of oculomotor behavior]. AB - The oculomotricity rests on complex anatomo-physiological bases and allows the binocular vision, which is a sensory function very completed in the human. After a short embryologic recall, the binocular vision is defined. The anatomical structures, which take part in the binocularity, are reviewed within three functional stages: an effector peripheral stage (eye ball, extrinsic muscles and their nerves), a sub cortical central stage, generator of the movements (oculomotor nuclei, reticularis formation...), and a central, cortical and subcortical stage, incitator of the movements (cortical areas, vestibular nuclei, cerebellum...). These anatomical bases make it possible to understand the many disorders observed in pathology. PMID- 15547479 TI - [Childhood refraction: epidemiology, progression, evaluation and a method for correcting ametropia]. AB - Children's refraction and its progression are important because refractive errors can be precisely corrected. Some types of simple ametropia can generate reversible perturbations after optical correction and others are amblyogenic and strabogenic ametropia. Definition, epidemiology (frequency and risk factors), progression of refraction, evaluation and an optical correction method are described. PMID- 15547481 TI - [Ocular manifestations of Down syndrome and cytogenic aspects]. PMID- 15547482 TI - [Cost-benefit evaluation of spinal cord stimulation treatment for failed-back surgery syndrome patients]. AB - BACKGROUND AND PURPOSE: Spinal cord stimulation is a well-known treatment of rigorously selected failed-back surgery syndrome patients. Efficacy levels over 50% of pain relief have been reported in long-term studies. The objective of this multicenter prospective evaluation was to analyze the cost to benefit ratio of spinal cord stimulation treatment for failed back surgery syndrome patients. METHODS: Nine hospitals (pain evaluation and treatment centers) were involved in the study. Forty-three patients were selected and implanted between January 1999 and January 2000. For each patient, pre- and post-operative evaluations (6, 12 and 24 months after implantation) were performed to assess pain relief and economical impact on pain treatment costs. RESULTS: After 24 months, mean 60% pain relief was achieved as assessed with the neuropathic pain score using a Visual Analog Scale (success rate=70%), whereas low-back pain was moderately reduced (29%). The Oswestry Disability questionnaire score was improved by a mean 39%. Costs of pain treatment (medication, consultation, other) are reduced by a mean 64% (1705 Euro) per patient per year. CONCLUSIONS: This study confirms a clear analgesic effect on neuropathic sciatalgia, and moderate attenuation of low back pain. One particular interest of this study is the medico-economic prospective evaluation showing that the initial cost of the implanted device is compensated by a significant, early, and stable reduction in the cost of associated pain therapies. PMID- 15547483 TI - [Aneurysmal rupture during embolization with Guglielmi detachable coils]. AB - OBJECTIVE: The purpose of this study is to determinate the frequency, causes, management and outcome of aneurysmal rupture occurring during embolization. We present our experience with this severe and feared complication. METHODS: We retrospectively reviewed 314 acute cerebral aneurysms that were treated with endovascular coiling. These patients were identified and the management and outcomes were recorded. The literature was reviewed. RESULTS: Six patients had an intraprocedural aneurysmal rupture. This complication occurred sporadically. Prevalence was 1.9%. Of these six, four were women and two were men. The mean age was 68 years (range: 43-74 years). Four aneurysms were located in the anterior circulation and two in the posterior circulation. Perforation occurred during microcatheterization of the aneurysm in one case and during coil deposition in five cases. In these five patients, aneurysmal rupture resulted from detachment of the first coil in three patients and detachment of the third and last coil in two patients. Hemodynamic changes were noted for one patient. The Glasgow Outcome Scale score at last follow-up examination was 1 in three patients and 3 in one patient (fair recovery). Mortality was 33% and morbidity was 16.7%. CONCLUSION: Aneurysmal perforation during embolization is a rare event (1.8 to 4.4%). When perforation is recognized, embolization can be completed immediately with further coil deposition and reversal of anticoagulation therapy. PMID- 15547484 TI - [Hydroxyurea treatment for unresectable meningioma]. AB - BACKGROUND: Management of unresectable progressive meningioma remains controversial and constitutes a major challenge since therapeutic options including chemotherapy and hormone modulation are limited. Recent data have suggested that hydroxyurea treatment may have an antitumoral effect. The purpose of this prospective phase II study was to evaluate the efficacy of hydroxyurea treatment for unresectable progressive meningioma. METHODS: From 1997 to 1999, consecutive patients presenting unresectable meningioma with clinically and/or neuroradiologically documented progression were considered for entry into this protocol. Previous radiotherapy was not a mandatory inclusion criteria. Treatment consisted of continuous oral administration of hydroxyurea at a dose of 20 mg/kg per day. Follow-up assessment included physical examination, computed tomography (CT), and magnetic resonance imaging (MRI) performed every three months, as well as regular blood testing. The primary endpoint was documentation of objective response by MRI or CT. RESULTS: The intent-to-treat population was 43 patients with at least 18 months follow-up. Median age was 60.4 years. Twenty-eight patients had undergone surgery following initial diagnosis. The meningioma was located in the skull base in 67% of patients. Histology was benign in 18 and atypical in 10. The eligible population included 36 patients with documented progressive disease at the time of inclusion; with progression documented clinically in 29 (67.5%) and/or radiologically in 20 (46%). In 7 patients, clinical or radiological progression could not be confirmed. The intent-to-treat analysis at median 26 months follow-up revealed objective response to hydroxyurea in only 3 patients (7%) including one on the basis of improvement in visual symptoms and two on MRI analysis. Progressive disease was observed clinically or radiologically in 26 patients (60.5%). Of the eligible population (n=36), 2 achieved an objective response and 13 (36%) exhibited stabilization under hydroxyurea therapy, while 21 (58%) progressed under treatment. Overall tolerance was good but anemia (grade I-II) and asthenia (grade I-II) were observed in 28% and 23.5% respectively. Treatment was discontinued in 3 patients because of chronic skin toxicity in one and anemia and asthenia in two. CONCLUSION: Hydroxyurea treatment is of marginal efficacy for meningioma and must not be considered as an alternative if radiotherapy or surgery is feasible. New efficient medical treatments are still required for progressive meningiomas. PMID- 15547485 TI - [Contribution of PET to the management of patients with low-grade glioma]. AB - BACKGROUND AND PURPOSE: Management of patients with low-grade glioma is a major challenge for the neurosurgeon. When is neurosurgery indicated? Should chemotherapy or radiotherapy be used? Many questions without an answer. We reviewed our experience with 65 patients treated for low-grade glioma who had preoperative PET images (FDG or/and MET). We examined the prognostic value of PET and also determined the sensitivity and the specificity of PET images to predict outcome. METHODS: Sixty-five patients with a FDG or MET PET images were analyzed. We used two visual scales and had complete follow-up data for 63 patients. The free interval was the principal criterion for statistical analysis. The sensitivity and the specificity of PET images was determined. RESULTS: Strong FDG uptake was correlated with a short free interval (p=0.001). Similar results were found with the MET analysis (p=0.0076). We had a PET with MET and FDG for 36 patients. The sensitivity was 66% and the specificity 94% for FDG PET. Sensitivity was 100% and specificity 53% for MET PET. CONCLUSIONS: PET imaging provides a prognostic factor independent from histology. MET PET is the best exam for the follow-up of patients with low-grade glioma and is helpful for separating aggressive from low-grade glioma. PMID- 15547486 TI - [Peroperative functional mapping using direct electrical stimulations. Methodological considerations]. AB - The interindividual anatomo-functional variability of the central nervous system implies that brain surgery within eloquent regions may induce neurological sequelae. Consequently, several methods of functional mapping were developed, both preoperative non-invasive neurofunctional imaging and intraoperative cortico subcortical electrical stimulations. While this technique was reported as safe, accurate, reliable and reproducible in the recent literature, a rigorous methodology is nevertheless mandatory in order to avoid any error in the detection of the so-called eloquent structures. Indeed, an erroneous mapping could lead to prematurely interrupting the resection (false positive), or to generate a postoperative permanent deficit due to the removal of critical areas interpreted as non-essential to function (false negative). The goal of the present review is to recall the electrophysiological principles of direct brain stimulations, and to consider the selection of stimulation parameters according to a theoretical approach, in order to adapt in practice the methods to each patient. The results reported in the literature are then analyzed, concerning the clinico-surgical contribution of intraoperative electrical mapping (in terms of extent of surgical indications, minimization of risk of permanent deficit, and quality of resection), their methodological interest (e.g. validation of neurofunctional imaging techniques), and their contribution to neurosciences (better understanding of the pathophysiology of brain structures, of the connectivity, and of the dynamic mechanisms underlying plasticity). PMID- 15547487 TI - [Contribution of percutaneous acrylic vertebroplasty for tumor-related spinal cord compression]. AB - We report two cases of spinal cord compression due to vertebral hemangioma in the first patient and myeloma in the other. Vertebroplasty was performed in both patients, combined with surgery for the first and radiotherapy for the second. Postprocedural pain decreased within 24 hours. Indications for percutaneous vertebroplasty are limited to aggressive vertebral hemangiomas, malignant spinal tumors including metastasis, multiple myeloma and solitary bone plasmocytoma. Vertebroplasty consolidates the vertebral body and reduces the risk of hemorrhage. It allows provides a rapid analgesic effect, radicullomedullary decompression associated with neurosurgery, spinal stabilization and can restrict progression of vascular malformations or tumors. Incidents and complications appear to be rare. PMID- 15547488 TI - [Persistent primitive trigeminal artery associated with brain cavernoma. Case report]. AB - Among cases of embryonic carotid-basilar anastomosis which may persist after birth, persistent trigeminal artery is the most common. It has been associated with a wide variety of intracranial abnormalities. We are unaware of any other reported association with cavernoma. We report a young woman who experienced seizures following spontaneous abortion. A CT scan disclosed a right frontal hematoma. MRI revealed a cavernoma associated with a persistent trigeminal artery. The cavernoma was removed through a frontal approach. The aim of the present case is to report another type of lesion fortuitously associated with a persistent trigeminal artery. PMID- 15547489 TI - Effect of over-expression of pigment epithelium derived factor (PEDF) on developing retinal vasculature in the mouse. AB - PURPOSE: Pigment epithelium derived factor (PEDF) is a secreted protein with demonstrated anti-angiogenic properties, and with potential application for the treatment of neovascular disease. Delivery of pigment epithelium derived factor to the retina via virus mediated gene transfer has been shown to inhibit neovascularization in a number of experimental models. While pigment epithelium derived factor is endogenously expressed in the retina, its role in guiding normal vessel development and growth is not yet known. This study aimed to determine whether over-expression of pigment epithelium derived factor alters the normal pattern of retinal vessel development. METHODS: Neonatal (age postnatal day 2 (P2)) CD1 mice were injected subretinally unilaterally with AAV2/1.CMV.PEDF while contralateral eyes were injected subretinally with AAV2/1.CMV.EGFP as control. Cohorts of animals were sacrificed at P7 to P21 and the retinal vasculature was co-labeled through fluorescein-dextran perfusion and immunohistochemistry. Vascular size, localization, and structure were analyzed using light and confocal microscopy. Additional cohorts were use to obtain quantitative levels of pigment epithelium derived factor protein through ELISA. RESULTS: The extent of vessel growth from the optic disk to periphery over time (i.e., the radius of retinal vasculature), and the area of expansion of the neural retina were unaffected by over-expression of pigment epithelium derived factor to levels at least 3.5 fold higher than endogenous levels. The thicknesses of the various retinal layers were similar in AAV2/1.CMV.PEDF treated and control injected eyes. Three dimensional analysis of confocal images shows a slight delay in the rate of growth of vasculature into the deeper layers of the retina in pigment epithelium derived factor treated eyes compared to EGFP treated control eyes. However, the normal differentiation of vessels into arterioles, and venules, and the formation of a capillary network continued to occur, achieving normal and complete maturation of vascular structure by P21. CONCLUSIONS: Over expression of pigment epithelium derived factor in the developing retina exerted no marked or permanent effects on retinal vessel growth and differentiation. The findings are relevant to the safety of the potential therapeutic use of pigment epithelium derived factor in human retinal disease. PMID- 15547490 TI - Alkali burn causes aldehyde dehydrogenase 3A1 (ALDH3A1) decrease in mouse cornea. AB - PURPOSE: Aldehyde dehydrogenase 3A1 (ALDH3A1) is the most abundant soluble protein component in the mouse cornea, produced mainly by corneal epithelial cells. High levels of ALDH3A1 in cornea contribute to maintenance of a stable an d transparent corneal structure. Alkali burn is a common damage to the corneal surface, which produces an alkaline hydrolysis of matrix proteins and induces an inflammatory reaction. Our study was intended to detect changes in ALDH3A1 expression after corneal alkaline burn. METHODS: To address this issue we employed RTQ-PCR to monitor the transcriptional change of ALDH3A1 after alkali burn. We used zymography to test enzyme activity changes of ALDH3A1 in the alkali burn cornea; And SDS-PAGE and mass spectrometry technology were used to verify protein content changes and to identify ALDH3A1 protein. RESULTS: Using zymography, ALDH3A1 enzymic activity was observed to decrease immediately after corneal alkali burn and the levels recovered following healing. Proteins extracted from alkali burned corneas, when run on SDS-PAGE, showed the same sized band (about 54 kDa, which is the molecular weight of ALDH3A1) but in much smaller quantity, compared to normal corneas. This result was further verified by mass spectrometry fingerprinting of the in-gel lysis product. An immediate decrease of ALDH3A1 transcription after alkali burning of the cornea was also found using RTQ PCR. This level of transcription was gradually restored during healing. CONCLUSIONS: Alkali burn of the corneal surface caused a rapid decrease of ALDH3A1 in the corneal at both the RNA and protein levels, which leads to the loses of the protective component of the corneal surface and makes it vulnerable to further damage. The ALDH3A1 level in the cornea gradually recovered during the healing process. Use of an anti-oxidation reagent as a treatment ingredient for alkali burn of the corneal surface could compensate for the decrease of anti oxidation protection potential caused by ALDH3A1 loss. PMID- 15547491 TI - Absence of myocilin and optineurin mutations in a large Philippine family with juvenile onset primary open angle glaucoma. AB - PURPOSE: To analyze the role of the two primary open angle glaucoma (POAG) genes, myocilin (MYOC) and optineurin (OPTN), in a large Philippine family segregating autosomal dominant juvenile onset open angle glaucoma (JOAG). METHODS: The coding sequences of the MYOC and OPTN genes were screened in 27 family members by polymerase chain reaction and direct sequencing. The specific MYOC promoter polymorphism (MYOC.mtl) was identified by restriction endonuclease assay. All of the ABI MD-10 microsatellite markers on chromosomes 1, 2, 3, 7, 8, and 10, which harbor the six known POAG loci, were analyzed for linkage with POAG. RESULTS: No mutation was identified in this large kindred. Instead, three polymorphisms (-80G >A, -1000G->C, R76K) in MYOC and four polymorphisms (T34T, M98K, R545Q, IVS7+24G >A) in OPTN were found. All markers flanking the six known POAG loci gave LOD scores not more than 1.1. Non-parametric linkage analysis for all these markers resulted in p values more than 0.05. CONCLUSIONS: Both mutation testing and linkage analysis provide strong evidence against MYOC and OPTN being the causative gene in this large family. It indicates that unidentified genes will underlie the occurrence of glaucoma in this family. PMID- 15547492 TI - Gene expression profile in corneal neovascularization identified by immunology related macroarray. AB - PURPOSE: To identify differentially expressed genes in corneal neovascularization using cDNA macroarray. METHODS: Mechanical denudation of corneal and limbal epithelium was performed to induce corneal vascularization in mice. Corneas were harvested 4 days after operation. Total RNA was isolated from both normal and vascularized corneas and used for the synthesis of cDNA probes. 32P labeled exponential cDNA probes were hybridized to mouse cDNA immunology arrays. To validate the gene expression patterns revealed by the cDNA expression array analysis, semiquantitative reverse transcription-polymerase chain reaction (RT PCR) and statistical analysis were performed to compare the normal and vascularized corneal samples. RESULTS: Of 545 immunology related genes on the arrays, 6 genes were upregulated and 1 gene was downregulated in the vascularized corneas compared with the normal corneas. Semi-quantitative RT-PCR was performed on the 6 genes selected in the arrays study, and showed that calreticulin (calregulin), apolipoprotein E, HSP84 (HSP90beta), and pleiotrophin were upregulated while interferon regulatory factor-1 was downregulated in the vascularized corneas compared with the normal corneas. These genes possess a number of biological functions including molecular chaperon, growth factor, and transcriptional factors. CONCLUSIONS: The differentially expressed genes newly identified in the context of corneal neovascularization represent novel candidate factors for further functional studies of the mechanisms of corneal neovascularization. Our data may provide new insight into the biological process of inflammation induced corneal neovascularization. PMID- 15547494 TI - The emergency response triangle. PMID- 15547495 TI - A cry for help. PMID- 15547496 TI - Enjoying the ride. PMID- 15547497 TI - Traction splints: more training required. PMID- 15547498 TI - Count to 10 before you discipline. PMID- 15547499 TI - One wheel at a time. How to flatten those bumps. PMID- 15547501 TI - The 'Brady-PEA' puzzle. PMID- 15547503 TI - Complexities of blunt chest trauma. PMID- 15547505 TI - DMATs ordered to Hurricane Charley. PMID- 15547504 TI - Dancing with Charley & Frances. Behind the scenes of storm management. PMID- 15547506 TI - Diversion in a state of emergency. PMID- 15547507 TI - EMS champions of injury prevention. Highlights from some of the best injury prevention programs in the United States. PMID- 15547517 TI - Elective induction of labor: should we make a fuss? PMID- 15547518 TI - Cesarean delivery after elective induction in nulliparous women: the physician effect. AB - OBJECTIVE: The purpose of this study was to assess the contribution of the individual physician to the probability of cesarean delivery among nulliparous women who undergo elective induction. STUDY DESIGN: A cohort study compared spontaneous labor and elective induction for all term, singleton, and cephalic nulliparous gestations over 2 years (1999-2000) at a large metropolitan hospital (n = 3215). Bivariate analysis was used to identify significant variables that were related to cesarean delivery. Logistic regression analysis was used to calculate the adjusted risk of cesarean delivery and to assess the independent effect of individual physician practice on cesarean delivery. Physician effect was evaluated in terms of its relative contribution to the explanatory power of the logistic regression model to predict cesarean delivery. RESULTS: The adjusted odds ratio for cesarean delivery with elective induction was 1.78 (95% CI, 1.39, 2.27). Using forward stepwise regression, the individual physician was a significant independent risk factor in the model with an R square statistic increase from 0.194 to 0.249 when the physician statistic was added to the final model. CONCLUSION: Nulliparous women are at a significant increased risk of cesarean delivery if elective induction is performed. The individual physician has a contributing effect to this increased risk. PMID- 15547519 TI - Active management of risk in pregnancy at term in an urban population: an association between a higher induction of labor rate and a lower cesarean delivery rate. AB - OBJECTIVE: The purpose of this study was to determine whether exposure to an alternative method of care, called the active management of risk in pregnancy at term, was associated with a lower group cesarean delivery rate. STUDY DESIGN: Active management of risk in pregnancy at term used risk factors for cesarean delivery to guide an increased use of labor induction. A retrospective cohort design was used to compare clinical outcomes of 100 pregnant women who were exposed to active management of risk in pregnancy at term to 300 randomly selected subjects who received standard management. RESULTS: The 2 groups had comparable levels of prenatal risk. The group exposed to the active management of risk in pregnancy at term exposure group encountered a higher induction rate (63% vs 25.7%; P < .001) and a lower cesarean delivery rate (4% vs 16.7%; P = .01). Findings were similar for both nulliparous and multiparous subgroups. Active management of risk in pregnancy at term exposure was not associated with higher rates of other major birth outcomes. CONCLUSION: Exposure to the active management of risk in pregnancy at term exposure was associated with a significantly lower group cesarean delivery rate. A prospective randomized trial that involved active management of risk in pregnancy at term exposure is needed to further explore this association. PMID- 15547520 TI - Fetoscopic surgery in triplet pregnancies: a multicenter case series. AB - OBJECTIVE: The purpose of this study was to evaluate the feasibility of fetoscopic surgery in complicated monochorionic and dichorionic triplet pregnancies. STUDY DESIGN: This was a case series of fetoscopic surgery in 12 cases of monochorionic (n = 5) or dichorionic (n = 7) triamniotic triplets complicated by severe feto-fetal transfusion (n = 8), twin reversed arterial perfusion sequence (n = 3), or discordant for an anomaly (n = 1). Six underwent cord occlusion, and 6 had laser coagulation of the vascular anastomoses. RESULTS: All procedures were successfully completed, and the outcome was comparable to that reported for similar procedures in twins. Eighty-three percent of fetuses survived, and 13% had long-term problems. CONCLUSION: Fetoscopic surgery may be considered as a therapeutic option in complicated triplet pregnancies, with results similar to those found in twins. PMID- 15547521 TI - Risk factors for prolapse recurrence after vaginal repair. AB - OBJECTIVE: The purpose of this study was to determine factors that are associated with recurrent prolapse. STUDY DESIGN: Of 389 women who underwent vaginal prolapse and incontinence between June 1996 and May 1999, 176 women had 1-year follow-up evaluations. Recurrent prolapse was analyzed by both pelvic organ prolapse quantification stage and centimeter measurements that were relative to the hymen. Logistic regression was used to determine odds ratios and 95% CI for factors that were associated with recurrent prolapse. RESULTS: One year after surgery, 102 women (58%) had recurrent prolapse (>/=stage II). Seventeen women (10%) had prolapse >/=1 cm beyond the hymen. Age <60 years (odds ratio, 3.2; 95% CI, 1.6-6.4; P = .001) and preoperative pelvic organ prolapse quantification stage III or IV (odds ratio, 2.7; 95% CI, 1.3-5.3; P = .005) were associated with a greater likelihood of recurrent prolapse (>/=stage II) at 1 year. CONCLUSION: Younger women and women with more advanced prolapse are more likely to experience recurrent prolapse after vaginal repair. PMID- 15547522 TI - Is rectovaginal endometriosis a progressive disease? AB - OBJECTIVE: The purpose of this study was to observe the natural history of untreated asymptomatic rectovaginal endometriosis. STUDY DESIGN: This was a prospective, observational study. Eighty-eight patients with untreated asymptomatic rectovaginal endometriosis were followed for 1 to 9 years. Pain symptoms and clinical and transrectal ultrasonographic findings were evaluated before and every 6 months after diagnosis. RESULTS: Two patients had specific symptoms that were attributable to rectovaginal endometriosis that was associated with an increase in lesion size and underwent surgery. In 4 other patients, the size of the endometriotic lesions increased, but the patients remained symptom free. The estimated cumulative proportion of patients with progression of disease and/or appearance of pain symptoms that were attributable to rectovaginal endometriosis after 6 years of follow up was 9.7%. For the remaining patients, the follow-up period was uneventful, with no detectable clinical nor echographic changes of the lesions and with no appearance of new symptoms. CONCLUSION: Progression of the disease and appearance of specific symptoms rarely occurred in patients with asymptomatic rectovaginal endometriosis. PMID- 15547523 TI - Gender selection: pressure from patients and industry should not alter our adherence to ethical guidelines. AB - Patients who undergo assisted reproduction occasionally request that physicians intervene with techniques that help to determine the gender of their offspring. Conventionally, all of these methods require an invasive procedure that places both the mother and pregnancy at risk. However, for sex-linked disorders, the risk/benefit ratio is favorable; therefore, in such cases, gender selection is warranted. The recent introduction of noninvasive techniques for X- and Y chromosome-bearing sperm sorting now provides another option for couples. However, the method is not absolute in its ability to sort sperm correctly; in many cases, the offspring are not of the desired sex. Sperm sorting has been marketed increasingly as a means for "family balancing," which is contrary to recommendations that are offered by ethics committees of several professional societies. More studies are needed with respect to the impact of gender selection on families before this method is introduced into routine practice. PMID- 15547524 TI - Condition-specific antepartum fetal testing. AB - OBJECTIVE: The purpose of this study was to determine the best available antepartum fetal testing methods according to the underlying pathophysiologic condition. STUDY DESIGN: We reviewed the current literature and our clinical experience with respect to condition-specific antepartum fetal testing. RESULTS: The efficacy of most antepartum tests that we use today is not supported by randomized controlled clinical trials, but from observational nonrandomized studies and expert opinion (evidence levels II or III). CONCLUSION: Based on the available evidence, the accuracy of a test depends on the underlying pathophysiologic condition. To improve accuracy, we must use condition-specific fetal testing. PMID- 15547525 TI - The epigenetics of ovarian cancer drug resistance and resensitization. AB - Ovarian cancer is the most lethal of all gynecologic neoplasms. Early-stage malignancy is frequently asymptomatic and difficult to detect and thus, by the time of diagnosis, most women have advanced disease. Most of these patients, although initially responsive, eventually develop and succumb to drug-resistant metastases. The success of typical postsurgical regimens, usually a platinum/taxane combination, is limited by primary tumors being intrinsically refractory to treatment and initially responsive tumors becoming refractory to treatment, due to the emergence of drug-resistant tumor cells. This review highlights a prominent role for epigenetics, particularly aberrant DNA methylation and histone acetylation, in both intrinsic and acquired drug resistance genetic pathways in ovarian cancer. Administration of therapies that reverse epigenetic "silencing" of tumor suppressors and other genes involved in drug response cascades could prove useful in the management of drug-resistant ovarian cancer patients. In this review, we summarize recent advances in the use of methyltransferase and histone deacetylase inhibitors and possible synergistic combinations of these to achieve maximal tumor suppressor gene re-expression. Moreover, when used in combination with conventional chemotherapeutic agents, epigenetic-based therapies may provide a means to resensitize ovarian tumors to the proven cytotoxic activities of conventional chemotherapeutics. PMID- 15547526 TI - Acupuncture for labor pain management: A systematic review. AB - Acupuncture is widely used to alleviate symptoms in a variety of painful conditions. In obstetrics and gynecology, acupuncture has also been applied to a range of conditions including labor pain. This systematic review aims to critically evaluate the evidence on analgesic effect of acupuncture during labor. Computerized literature searches of 7 databases were performed for randomized clinical trials (RCTs) of acupuncture involving needle insertion for pain during labor. Three RCTs were identified and their methodologic quality was generally good. Two RCTs compared adjunctive acupuncture with usual care only and reported a reduction of meperidine and/or epidural analgesia. One placebo acupuncture controlled trial showed a statistically significant difference in both subjective and objective outcome measures of pain. No adverse events were reported in any of the trials. It is concluded that the evidence for acupuncture as an adjunct to conventional pain control during labor is promising but, because of the paucity of trial data, not convincing. Further research is warranted to clearly define its place in labor pain management. PMID- 15547527 TI - Does metformin induce ovulation in normoandrogenic anovulatory women? AB - OBJECTIVE: This study was undertaken to evaluate the efficacy of metformin in women with anovulation who do not have evidence for hyperandrogenism and classic polycystic ovary syndrome. STUDY DESIGN: A randomized trial of metformin (1500 mg daily) and placebo in 24 anovulatory women was undertaken for 3 months. Assessments of changes in hormone levels and insulin sensitivity were carried out. Abnormal hormonal values were defined by levels exceeding the range in normal ovulatory controls. RESULTS: Anovulatory women had normal androgen levels and luteinizing hormone but had higher serum insulin and lower insulin sensitivity compared with controls. Over 3 months, there were 16 ovulatory cycles with metformin and only 4 with placebo ( P < .05). Success of ovulation did not correlate with changes in androgen, insulin, or insulin sensitivity parameters. CONCLUSION: Metformin may be useful for inducing ovulation in anovulatory women who do not have hyperandrogenism. This effect may be independent of a lowering of androgen or insulin levels. PMID- 15547528 TI - NovaSure impedance controlled system for endometrial ablation: three-year follow up on 107 patients. AB - OBJECTIVE: This study was undertaken to assess the safety, efficacy, and data durability of the NovaSure ablation at 3 years after the procedure in women with menorrhagia secondary to dysfunctional uterine bleeding (DUB). STUDY DESIGN: A prospective, single-arm, observational pilot study (Canadian Task Force classification II-1) was carried out at a specialized center for gynecologic endoscopy with 107 premenopausal women with menorrhagia secondary to DUB. NovaSure ablation was performed in 107 patients. Pictorial Blood loss Assessment Chart diary sampling was used to assess menstrual blood loss. Ablation was performed without any type of endometrial pretreatment. RESULTS: No intraoperative or postoperative complications were observed. Treatment time averaged 94 seconds; 65% of the patients reported amenorrhea. Hysterectomy was avoided in 97.2% of patients at 3-year follow-up. CONCLUSION: Long-term clinical results demonstrate that the NovaSure system is a safe and effective method for treatment of women with menorrhagia secondary to DUB, yielding high amenorrhea and success rates, with low re-treatment rates. PMID- 15547529 TI - Hybrid capture 2 viral load and the 2-year cumulative risk of cervical intraepithelial neoplasia grade 3 or cancer. AB - OBJECTIVE: The purpose of this study was to determine the clinical value of a semiquantitative measure of human papillomavirus viral load by the hybrid capture 2 assay for stratification of the risk of histologic cervical intraepithelial neoplasia grade 3 or carcinoma. STUDY DESIGN: The Atypical Cells of Unknown Significance and Low-Grade Squamous Intraepithelial Lesions Triage Study was a randomized clinical trial of 5060 women with 2 years of follow-up to evaluate treatment strategies for women with equivocal or mildly abnormal cervical cytologic condition. The usefulness of the continuous hybrid capture 2 output relative light units/positive controls that were above the positive threshold (1.0 relative light units/positive controls), which was a surrogate for human papillomavirus viral load, for distinguishing between hybrid capture 2 positive women who were diagnosed with cervical intraepithelial neoplasia grade 3 or carcinoma during the study from those who were not diagnosed with cervical intraepithelial neoplasia grade 3 or carcinoma was examined with the use of receiver-operator characteristic analyses. RESULTS: Relative light units/positive controls values did not further discriminate between hybrid capture 2 positive women with cervical intraepithelial neoplasia grade 3 or carcinoma from those with less than cervical intraepithelial neoplasia grade 3 or carcinoma. The use of a cervical intraepithelial neoplasia grade 2 or more severe or carcinoma case definition did not alter our findings. CONCLUSION: Among women with atypical cells of unknown significance or low-grade squamous intraepithelial lesion cervical cytologic findings, the hybrid capture 2 viral load measurement did not improve the detection of 2-year cumulative cases of cervical intraepithelial neoplasia grade 3 or carcinoma significantly. PMID- 15547530 TI - Interferon-gamma expression is an independent prognostic factor in ovarian cancer. AB - BACKGROUND: Epithelial ovarian cancer prognosis is improved by the presence of intratumoral CD3 + T cells, which are known to produce interferon-gamma. We therefore speculated that interferon-gamma expression in ovarian cancer infiltrating T-lymphocytes might cause better prognosis. PATIENTS AND METHODS: Reverse transcriptase polymerase chain reaction was performed to measure the expression of interferon-gamma and other related genes in normal ovaries (n = 19) and in ovarian cancer specimens (n = 99). Median follow-up of patients was 5.8 years. RESULTS: Interferon-gamma and CD-3 expression did not significantly differ in normal and malignant tissue. Patients with high levels of interferon-gamma expression had significantly longer progression-free and overall survival. Median time to progression was 10 and 29 months for patients with low and high interferon-gamma expression, respectively ( P = .039). Corresponding survival times were 29 and 44 months ( P < .032). Application of multivariate Cox regression analysis showed interferon-gamma expression to be an independent prognostic factor for progression-free and overall survival. CONCLUSION: Elevated interferon-gamma expression correlates with improved clinical outcome in patients with ovarian cancer. PMID- 15547531 TI - The effects of repeated spectroscopic pressure measurements on fluorescence intensity in the cervix. AB - OBJECTIVE: Fluorescence spectroscopy is a promising technology for the detection of cervical squamous intraepithelial lesions (SILs). In this study we took repeated measures in the cervix to determine whether the order of measurement produces changes in fluorescence intensity and whether there are differences in variation due to pressure. METHODS: A pressure sensitive fiber-optic probe to measure fluorescence spectra was calibrated at light, medium, and firm levels (0.2, 0.4, 0.6 N). Measurements were made 3 times at each of 2 sites in the patient's cervix. Spectroscopic data were preprocessed and analyzed to compare order of pressure and intensity variability as a function of pressure on measurements. RESULTS: Four providers took 3 measurements from 2 sites each in 18 patients, yielding 108 measurements. After corrections for multiple comparisons, neither the order of probe pressure nor the variability of probe pressure significantly affected variations in fluorescence intensity. CONCLUSION: This study shows that the probe pressure variability is probably not an issue for these devices. PMID- 15547532 TI - The effect of number of throws on knot security with nonidentical sliding knots. AB - OBJECTIVE: The study was undertaken to test the integrity of nonidentical sliding knots made with 3 throws compared with those made with 6 throws with monofilament and braided absorbable suture. STUDY DESIGN: The 3 throw nonidentical sliding knot was compared with the 6 throw nonidentical sliding knot in 4 different suture groups. The groups were 0-0 polydioxanone, 2-0 polydioxanone, 0-0 polyglactin 910, and 2-0 polyglactin 910. Knots were tested to failure with a tensiometer. The proportion of 3 throw knots becoming untied was compared with the 6 throw knot within each group. Ultimate load required to break tied knots within each suture group was also evaluated. RESULTS: The 3 throw knots had very high rates of knot failure and untied significantly more often than the 6 throw knots. CONCLUSION: The 6 throw nonidentical sliding knot demonstrates superior knot integrity compared with the 3 throw knot with both monofilament and braided absorbable suture. PMID- 15547533 TI - Gene therapy of uterine leiomyomas: adenovirus-mediated expression of dominant negative estrogen receptor inhibits tumor growth in nude mice. AB - BACKGROUND: Leiomyomas (fibroids) are common estrogen-dependent uterine tumors with no effective medicinal treatment; hysterectomy is the mainstay of management. STUDY DESIGN: This study was undertaken to investigate a potential therapy for leiomyoma; we used a mutated dominant-negative estrogen receptor gene delivered via an adenoviral vector (Ad-ER-DN). RESULTS: Ad-ER-DN transduction, in both human and rat leiomyoma cell lines, induced an increase in both caspase-3 levels and BAX/Bcl-2 ratio with evident apoptosis in the TdT-mediated dUTP nick end labeling assay. In nude mice, rat leiomyoma cells ex vivo transduced with Ad ER-DN supported significantly smaller tumors compared with Ad-LacZ-treated cells 5 weeks after implantation. In mice treated by direct intratumor injection into preexisting lesions, Ad-ER-DN caused immediate overall arrest of tumor growth. The Ad-ER-DN-treated tumors demonstrated severely inhibited cell proliferation (BrdU index) and a marked increase in the number of apoptotic cells (TdT-mediated dUTP nick-end labeling index). CONCLUSION: Dominant-negative estrogen receptor gene therapy may provide a nonsurgical treatment option for women with symptomatic uterine fibroids who want to preserve their uteri. PMID- 15547534 TI - A randomized trial comparing a 30-mL and an 80-mL Foley catheter balloon for preinduction cervical ripening. AB - OBJECTIVES: The purpose of this study was to compare the efficacy of cervical ripening with 2 Foley catheter balloon volumes. STUDY DESIGN: Pregnant women admitted for induction of labor with a term singleton gestation were randomly assigned for cervical ripening by a balloon inflated with 30 mL or 80 mL of sterile saline. RESULTS: Two hundred and three women were included in the analysis. Ripening of the cervix with the larger balloon volume was associated with a significantly higher rate of post-ripening dilatation of 3 cm or more (76.0% vs 52.4%, P < .001). In primiparous women, the larger balloon volume resulted in a significantly higher rate of deliveries by 24 hours (71.4% vs 49%, P < .05), and a significantly less requirement of augmentation with oxytocin (69.3% vs 90.4%, P < .05). CONCLUSION: Ripening of the unfavorable cervix in primiparous women with a Foley catheter balloon inflated with 80 mL provided effective more dilatation, faster labor, and decreased need for oxytocin than with a balloon inflated with 30 mL of sterile saline. PMID- 15547535 TI - Labor induction with dinoprostone or oxytocine and postpartum disseminated intravascular coagulation: a hospital-based case-control study. AB - OBJECTIVE: This study was undertaken to test the hypothesis of an association between pharmacologic agents used for labor induction, in particular dinoprostone, and postpartum disseminated intravascular coagulation (DIC). STUDY DESIGN: A retrospective hospital-based case-control study. Adjusted odds ratios (AOR) were calculated by a conditional logistic regression. RESULTS: Forty valid cases of postpartum DIC were compared against 197 matched controls. Labor was induced in 17% of controls, and 56% of cases (AOR = 7.2; 95% CI: 2.1-24.6). The association was observed for both dinoprostone (AOR = 6.7; 95% CI: 1.7-26.5) and oxytocin (AOR = 8.4; 95% CI: 1.4-50.9). Other risk factors identified were as follows: a maternal age older than 34 years (AOR = 9.5; 95% CI: 2.4-37.7), complications during pregnancy (AOR = 5.5; 95% CI: 1.3-22.8), and a gestational age of over 40 weeks (AOR = 3.5; 95% CI: 1.1-11.1). Such factors were shown to also have an interaction with the induction of labor. Oxytocin augmentation showed a negative association (AOR = 0.1; 95% CI: 0.02-0.4). The absolute risk attributable to induction was estimated in 5 per 10,000 deliveries. CONCLUSION: The pharmacologic induction of labor is associated with an increased risk of postpartum DIC, regardless the substance used. Although the absolute risk seems to be quite low, the obstetricians should not neglect it, in particular for the special risk groups identified. PMID- 15547536 TI - Modified Bishop's score and induction of labor in patients with a previous cesarean delivery. AB - OBJECTIVES: The purpose of this study was to evaluate the association between preinduction modified Bishop's score and obstetric outcome, including successful vaginal birth after prior cesarean (VBAC) and uterine rupture in patients with a previous cesarean undergoing induction of labor. STUDY DESIGN: Medical records of all patients who had an induction of labor after a previous cesarean in our institution between 1988 and 2002 were reviewed. Patients were divided into 4 groups according to the modified Bishop's score (0 to 2, 3 to 5, 6 to 8, and 9 to 12). The rates of successful VBAC, symptomatic uterine rupture, and other obstetric outcomes were evaluated in each group. Multivariate regression analyses were performed to adjust for confounding factors. RESULTS: Out of 685 women included in the study, 187 (27.3%) had a modified Bishop's score <2, 276 (40.3%) of 3 to 5, 189 (27.6%) of 6 to 8, and 33 (4.8%) of 9 to 12. The rate of successful VBAC significantly correlated with the modified Bishop's score (57.5%, 64.5%, 82.5%, and 97.0%, respectively, P < .001). However, the rate of uterine rupture was not statistically significant between the groups (2.1%, 1.8%, 0.5%, 0.0%, P=.48). After adjusting for confounding variables, a modified Bishop's score >/=6 remained associated with successful VBAC (odds ratio [OR] 2.07, 95% CI 1.28-3.35, P < .001). CONCLUSION: The modified Bishop's score before induction of labor is an independent factor associated with successful VBAC. PMID- 15547537 TI - Interleukin-6, interleukin-8, and soluble tumor necrosis factor receptor-I in the cord blood as predictors of chronic lung disease in premature infants. AB - OBJECTIVES: In order to predict the late-development of chronic lung disease of prematurity (CLD), cytokines in the cord blood were assessed in this study. STUDY DESIGN: Eighteen premature infants with CLD were enrolled. Cord blood plasma levels of cytokines of these infants and 12 control infants without CLD were measured including interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-8, IL-10, interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, soluble TNF receptor I, and soluble IL-6 receptor using a cytometric bead array and an enzyme-linked immunosorbent assay. RESULTS: The cord blood IL-6, IL-8, and sTNFR-I levels were significantly elevated in CLD infants compared with those in control (P < .05). IL-1beta, IL-2, IL-4, IL-10, and IFN-gamma were undetectable in both groups. CLD infants with maternal chorioamnionitis had higher IL-6 than those without chorioamnionitis (P < .01). In CLD infants, IL-6 was higher in the infants who required prolonged oxygen therapy (P < .05). CONCLUSION: Elevated inflammatory cytokines in the cord blood are associated with the progression to CLD. PMID- 15547538 TI - The association between preeclampsia and the severity of gestational diabetes: the impact of glycemic control. AB - OBJECTIVES: We sought to determine if the rate of preeclampsia is related to the severity of gestational diabetes mellitus (GDM), and if it can be decreased by optimizing glycemic control. STUDY DESIGN: A retrospective analysis of prospectively collective data of 1813 patients with GDM was performed to determine the rate of preeclampsia. Patients were stratified after treatment was begun by level of glycemic control (well controlled was defined as mean blood glucose <95 mg/dL). The extent of hyperglycemia was analyzed by the level of the abnormality in the oral GTT and by the degree of abnormality of daily glucose control after treatment has begun. Severity of GDM was categorized using fasting plasma glucose (FPG) on a 3-hour oral GTT by 10 mg/dL increments. RESULTS: Overall, preeclampsia was diagnosed in 9.6% (174/1813) of diabetic patients. The GDM subjects who developed preeclampsia were significantly younger, had a higher nulliparity rate, were more obese, and gained significantly more weight during pregnancy. However, no difference was found in glycemic profile characteristics between the 2 groups. A comparison between patients with FPG <105 and FPG >105 revealed that the rate of preeclampsia increased significantly, 7.8% vs 13.8%, (O.R 1.81, 95%CI 1.3-2.51). For GDM patients with only mild hyperglycemia (FPG <105 mg/dL), no significant difference was found in the rate of preeclampsia. Preeclampsia rate was further evaluated in relation to level of glycemic control; for the well-controlled patients (mean blood glucose [MBG] <95 mg/dL, n=994), similar rates of preeclampsia were found between each category of FPG severity. In contrast, in poorly controlled patients (MBG >95 mg/dL, n=819), a comparison between severity threshold of FPG <115 and FPG >115 revealed that the preeclampsia rate was 9.8% vs 18% (O.R 2.56, 95%C.I. 1.5-4.3). In a logistic regression model, only prepregnancy BMI (O.R 2.3, 95%CI 1.16-2.30) and severity of GDM (O.R 1.7, 95%CI 1.21-2.38) were independently and significantly associated with an increased risk of preeclampsia. CONCLUSION: The rate of preeclampsia is influenced by the severity of GDM and prepregnancy BMI. Optimizing glucose control during pregnancy may decrease the rate of preeclampsia, even in those with a greater severity of GDM. PMID- 15547539 TI - Prophylactic antibiotics for non-laboring patients with intact membranes undergoing cesarean delivery: an economic analysis. AB - OBJECTIVE: We conducted an economic analysis of prophylactic antibiotic administration for elective cesarean delivery. STUDY DESIGN: We created a decision tree comparing the use of prophylactic antibiotics (cefazolin 1 gm intravenous) with no antibiotic use. We modeled the potential for anaphylaxis, and included outcomes of fever and endometritis. Outcome probabilities and effectiveness of antibiotic administration were derived from published sources. Costs are 2001 estimates derived from our hospital's accounting system. Sensitivity analyses were performed over the range of actual patient costs and 95% CI of the risk and probability estimates. RESULTS: Cost of an uncomplicated elective cesarean delivery was $1638.57. Fever evaluation added $125.91. Elective procedure complicated by endometritis cost $2327.29. Cefazolin administration cost $1.01. The following estimates were used: relative risk (RR) of endometritis with antibiotics was 0.18 (95% CI 0.07-0.45), fever 0.47 (95% CI 0.32-0.68), risk of endometritis without prophylaxis 4.8% (95% CI 0.9%-43%), and fever without prophylaxis 14.4% (95% CI 4%-33%). Cost of an average case without prophylaxis was $1683.72; prophylaxis reduced this to $1653.06. Sensitivity analysis over the ranges above still yielded cost savings. CONCLUSION: Administration of prophylactic antibiotics for elective cesarean delivery reduced costs by $30.66 per case, approximately 2% of the total cost. Prophylactic antibiotic administration results in cost savings for elective cesarean delivery. PMID- 15547540 TI - A prediction model for superimposed preeclampsia in women with chronic hypertension during pregnancy. AB - OBJECTIVE: Women with chronic hypertension are at increased risk for superimposed preeclampsia. We developed a prediction algorithm for superimposed preeclampsia using clinical and laboratory information that were measured early in pregnancy. STUDY DESIGN: A secondary analysis of data that were collected from 110 women with chronic hypertension who were enrolled in a trial of calcium supplementation was performed. Blood pressure, the renin-angiotensin system, and calcium metabolism were assessed at 12, 20, 28, and 36 weeks of gestation and 6 weeks after delivery. Multivariable logistic regression was used to develop the predictive model. RESULTS: Thirty-seven women had superimposed preeclampsia. The final model included systolic blood pressure, serum uric acid, and plasma renin activity, which were all measured at 20 weeks of gestation. Women with high systolic blood pressure (>140 mm Hg), elevated uric acid (>3.6 mg/dL), and low plasma renin activity (<4 ng/mL/hr) had an 86% probability of having superimposed preeclampsia. Women with 2 risk factors had a 62% probability of superimposed preeclampsia, and women with only 1 risk factor had a 30% to 40% probability of superimposed preeclampsia. CONCLUSION: We developed a prediction algorithm that can be validated in future studies for superimposed preeclampsia for women with chronic hypertension. PMID- 15547541 TI - Accidental fetal lacerations during cesarean delivery: experience in an Italian level III university hospital. AB - OBJECTIVE: The purpose of this study was to investigate the incidence, type, location, and risk factors of accidental fetal lacerations during cesarean delivery. STUDY DESIGN: Total deliveries, cesarean deliveries, and neonatal records for documented accidental fetal lacerations were reviewed retrospectively in our level III university hospital. The gestational age, the presenting part of the fetus, the cesarean delivery indication, the type of incision, and the surgeon who performed the procedure were recorded. Cesarean deliveries were divided into scheduled, unscheduled, and emergency procedures. Fetal lacerations were divided into mild, moderate, and severe. Neonatal follow-up examinations regarding laceration sequelae were available for 6 months. RESULTS: Of 14926 deliveries, 3108 women were delivered by cesarean birth (20.82%). Neonatal records documented 97 accidental fetal lacerations. Of these accidental lacerations, 94 were mild; 2 were moderate, and 1 was severe. The overall rate of accidental fetal laceration per cesarean delivery was 3.12%; the accidental laceration rate in the cohort of fetuses was 2.46%. The crude odds ratios were 0.34 for scheduled procedures, 0.57 for unscheduled procedures, and 1.7 for emergency procedures. The risk for fetal accidental lacerations was higher in fetuses who underwent emergency cesarean birth and lower for unscheduled and scheduled cesarean births (P < .001). CONCLUSION: Fetal accidental laceration may occur during cesarean delivery; the incidence is significantly higher during emergency cesarean delivery compared with elective procedures. The patient should be counseled about the occurrence of fetal laceration during cesarean delivery to avoid litigation. PMID- 15547542 TI - The novel antimicrobial peptide beta3-defensin is produced by the amnion: a possible role of the fetal membranes in innate immunity of the amniotic cavity. AB - BACKGROUND: Innate immunity evolved to eliminate microorganisms before, or after their entry into the tissues, but before enough antigen is available to activate an adaptive, immune response. Innate immunity is so successful that the majority of encountered microbes are neutralized. The beta-defensins are antimicrobial peptides produced by skin and mucosal surfaces and are an integral part of the innate immune system. The ability of the amnion cells, which are epithelial derivatives, to produce antimicrobial beta-defensins has not been explored. OBJECTIVE: This study was undertaken to test the hypothesis that amnion cells synthesize beta-defensins under either basal or stimulated conditions. METHODS: Amnion epithelial FL cells (ATCC CCL 62) were cultured in Ham's F12 and Dulbecco's modified Eagle medium plus 10% fetal calf serum until confluence, then replated into 24-well plates at 1.5 million cells per well. Cells from triplicate wells were harvested after 1, 3, 6, and 24 hours of exposure to microbial wall components (lipopolysaccharide [LPS]: 1 microg/mL or peptidoglycan [PG]: 10 microg/mL). Reverse transcription real-time polymerase chain reaction was performed with the use of human-specific primers for beta1, beta2, beta3, and beta4 defensins to compare basal messenger RNA (mRNA) levels of defensins and in response to treatment. beta-actin was used for standardization. Protein expression was investigated by immunofluorescence of the cells in culture, and by immunohistochemistry in paraffin sections of human fetal membranes from pregnancies with or without histologic chorioamnionitis. RESULTS: Amnion FL cells expressed mRNA for all known beta-defensins with beta3-defensin mRNA levels significantly higher compared with others ( P < .001, 1-way analysis of variance [ANOVA]). beta3 was the only beta-defensin whose mRNA was upregulated in response to the microbial mimics LPS (1-way ANOVA, P = .019) and PG (1-way ANOVA, P = .011). Immunofluorescence confirmed that beta3-defensin protein was present in cultured amnion cells, and upregulated in response to PG and LPS in distinct cells. Similarly, in tissue sections of human fetal membranes amnion epithelium was intensely positive for beta3-defensin protein by immunohistochemistry. Conspicuous beta3-defensin staining was also detected in the chorio-decidua. CONCLUSION: Amnion cells have the ability to produce beta-defensins. The beta3 defensin appears to be the predominant epithelial defensin expressed. Its induction by microbial mimics suggests that the amniotic epithelium may play a role in the innate immunity of the amniotic cavity. PMID- 15547543 TI - Prenatal glucocorticoid exposure and postnatal adaptation in premature newborn baboons ventilated for six days. AB - OBJECTIVE: Renal and cardiovascular function is improved during the first 24 hours of life in preterm ventilated baboons exposed to prenatal betamethasone (BETA). We hypothesized BETA-induced effects would be sustained through day 6 of life. Study design Pregnant baboons received saline or BETA (6 mg) 48 and 24 hours before preterm delivery at 125 days' gestation. The newborn baboons were ventilated for 6 days, and assessed for renal, cardiovascular, and endocrine function. RESULTS: Mean arterial blood pressure (MAP) and glomerular filtration rate (GFR) values 24 hours after delivery were higher in the BETA group. Kidney Na, K-ATPase activity was higher in the BETA group by day 6. All other measures were similar in both groups by day 6. CONCLUSION: Prenatal BETA exposure in the premature baboon: (1) increases MAP and GFR on day 1 without measurable effects by day 6 and (2) increases kidney Na, K-ATPase activity. PMID- 15547544 TI - Physical and biomechanical characteristics of rat cervical ripening are not consistent with increased collagenase activity. AB - OBJECTIVE: The cervix progressively softens during pregnancy to allow stretch without rupture in labor. Cervical softening is the product of complex modifications that include increased proteoglycan-to-collagen ratio, increased hyaluronic acid and water content, and breakdown of collagen by matrix metalloproteases. The relative contribution of collagen breakdown to cervical ripening is unclear. We sought to identify, discriminate, and quantify the physical characteristics of rat cervix during pregnancy, labor, and both before and after exposure to either prostaglandin (PGE(2)) or the collagenolytic enzyme matrix metalloprotease-1 (MMP-1). STUDY DESIGN: Cervices were collected from nonpregnant rats in diestrus (n=4) and pregnant rats on d10 (n=4), d16 (n=11), d20 (n=5), and d22 (term) nonlabor (NL: n=4) and d22 in term labor (TL: n=7). Cervices were also collected from a separate group in preterm labor induced by RU486 (PTL: n=10). The effect of PGE(2) on cervical characteristics was determined after intravaginal placement of PGE(2) gel (0.5 mg PGE(2): n=3) or placebo metylcellulose gel (CRL(PG) n=6) for 20 hours before euthanasia on d16. The effect of collagen was determined by incubating in vitro cervices from untreated d16 rats with (MMP-1: n=3) and without (CRL(MMP): n=7) activated collagenase before tensile testing. Tensile properties were quantitated by using Shimadzu EZ-test instrumentation (Shimadzu North America, Columbia, Md) with a stretching regimen that mimicked labor contractions while recording the force opposed by the tissue. Parameters such as the slope (a measure of stiffness), yield point (YP; moment the tissue changes its proprieties from elastic to plastic), and break point (BP; a measure of tissue strength) were recorded and analyzed. The plateau was defined as the phase after YP but before BP. RESULTS: Compared with d16, cervical extensibility increased significantly by d20 (slope d16: 0.41 +/- 0.03 N/mm vs d20: 0.19 +/- 0.05 N/mm, P < .01), and during both PTL (slope: 0.17 +/- 0.03 N/mm) and TL (slope: 0.11 +/- 0.02 N/mm). This increase was mimicked by PGE(2) (slope PGE(2): 0.24 +/- 0.03 vs CRL(PG): 0.40 +/- 0.05 N/mm, P=.04), but not by collagenase (slope MMP-1: 0.35 +/- 0.02 vs CRL(MMP): 0.38 +/- 0.05 N/mm, P>.05). YP was significantly reduced as pregnancy advanced, whereas BP increased, suggesting both increased plasticity (compliance) and strength. However, the plateau length increased 3-fold both by d20 and after PGE(2). In contrast, the addition of MMP-1 reduced the plateau. BP occurred significantly earlier in collagenase-treated tissues, but later in PTL-, TL-, and PGE(2) treated cervices. CONCLUSION: The changes in physical properties of the rat cervix during physiologic ripening are similar to those induced by PGE(2) and RU486, and consist of increased extensibility, compliance, and strength. These changes cannot be attributed to increased collagenase activity, which would decrease tissue compliance and strength. PMID- 15547545 TI - Degranulation of uterine mast cell modifies contractility of isolated myometrium from pregnant women. AB - OBJECTIVE: This study was undertaken to test that uterine mast cell degranulation alters human myometrial contractility in vitro and to define what mediators are involved in this process. STUDY DESIGN: Longitudinal myometrial strips prepared from biopsy specimen obtained from the lower uterine segment of women at preterm and term gestation (with and without labor) were studied. Contractile responses to compound 48/80, a mast cell degranulator, were compared in the absence or presence of a mast cell stabilizer, H 1 and H 2 receptor antagonists, cyclooxygenase, and lipoxygenase inhibitors. RESULTS: Compound 48/80 increased myometrial contractility in all groups. The mast cell stabilizer cromolyn inhibited contractility, whereas the cyclooxygenase inhibitor ibuprofen, the H 1 receptor antagonist S(+)-chlorpheniramine maleate, but not the H 2 antagonist cimetidine, only slightly attenuated this effect. The lipoxygenase inhibitor linoleyl hydroxamic acid augmented the responses to compound 48/80 in the preterm but not in the term group. CONCLUSION: Uterine mast cell degranulation, or the effects of their mediators, can modulate uterine contractility during pregnancy. PMID- 15547546 TI - Sinogenic brain abscess complicating pregnancy. AB - Cerebral abscess is an extremely rare complication of pregnancy. We report a case of a patient at 36 weeks of gestation presenting with severe headache, confusion, and seizures after starting treatment for sinusitis. Imaging revealed a left temporal lobe abscess, which was treated with broad-spectrum antibiotics. Neurologic deterioration in a gravida with sinusitis suggests secondary sinogenic intracranial suppurative complications. PMID- 15547547 TI - Myoma expulsion after uterine artery embolization: complication or cure? AB - A 54-year-old woman had an expulsed myoma 10 weeks after uterine artery embolization. After treatment with antibiotics and a small surgical intervention, she recovered completely without any sign of myomatous disease afterwards. Patients should be informed about the possibility of expulsion. Expulsion of myomas after uterine artery embolization occurs relatively frequently and may be just one of the ways to attain cure. PMID- 15547548 TI - Successful surgical treatment of a primary liver gastrinoma during pregnancy: a case report. AB - Primary neuroendocrine tumors of the liver, particularly gastrinomas, are exceptionally rare. We present a case of a sporadic primary liver gastrinoma in a young woman in whom the definitive diagnosis was reached, and surgical therapy was performed during pregnancy. PMID- 15547549 TI - Vulvitis granulomatosa and unilateral hypertrophy of the vulva related to Crohn's disease: a case report. AB - We report the case of a patient with voluminous unilateral hypertrophy of vulvar labia and pseudocondylomata in the fossa navicularis associated with intestinal Crohn's disease. No fistulas were observed. The patient has had Crohn's disease for 8 years. Interrelationship of genital alterations and Crohn's disease are discussed. PMID- 15547550 TI - Acute gastric ulcer perforation in a 35 weeks' nulliparous patient with gastric banding. AB - We present a case of a primiparous patient at 35 weeks' gestation who had had laparoscopic gastric banding, and who presented to labor and delivery with protracted vomiting followed by an acute abdomen and fetal distress. An emergency surgery revealed acute gastric ulcer perforation. This complication, although rare, should be considered. PMID- 15547551 TI - Preeclampsia-like syndrome that is associated with severe hypothyroidism in a 20 week pregnant woman. AB - We report a case of overt hypothyroidism that was associated with a preeclampsia like syndrome and fetal death in a 37-year-old woman (gravidity, 7; parity, 6). Rapid and robust correction of hypothyroidism is recommended in this situation. PMID- 15547552 TI - Endometriosis of the liver containing mullerian adenosarcoma: case report. AB - We present a case of a liver endometrioma in a postmenopausal woman. After failed management with leuprolide acetate, the mass was resected and contained focal areas of mullerian adenosarcoma. This is a rare case of mullerian adenosarcoma that appeared to arise within an endometrioma of the liver. PMID- 15547553 TI - Hypogonadotropic hypogonadism in a female with the Johnson-McMillin syndrome. AB - A case of hypogonadotropic hypogonadism associated with the Johnson-McMillin syndrome is presented. This is a rare, autosomal dominant disorder, characterized by variable degrees of alopecia and anosmia, conductive hearing loss, and increased dental caries. Until now hypogonadotropic hypogonadism has only been observed in affected men. Ovulation can be induced with gonadotropins and conception can be obtained, but because prenatal diagnosis is not as yet possible, oocyte donation should be offered as an alternative for procreation. PMID- 15547554 TI - Use of maternal plasma for noninvasive determination of fetal RhD status. AB - Determination of the fetal RhD typing using free fetal DNA in maternal plasma is beginning to enjoy widespread acceptance in Europe. Case 1, the partner of an RhD sensitized patient, was identified with a heterozygous paternal phenotype by serologic testing. Maternal plasma was drawn at 18 weeks' gestation to determine the fetal RhD status. The result was unable to be reported as RhD negative; the patient subsequently underwent amniocentesis to confirm an RhD-negative fetus. Case 2, a partner of another RhD-sensitized patient, was similarly identified with a heterozygous paternal phenotype by serologic testing. Maternal plasma was also drawn at 18 weeks' gestation to determine the fetal RhD status. It returned RhD negative and allowed for the avoidance of invasive testing for the remainder of the pregnancy. Therefore, maternal plasma testing for fetal RhD status represents a new tool in the management of the cases of RhD alloimmunization in pregnancy. PMID- 15547555 TI - Leiomyosarcoma in a premenopausal patient after uterine artery embolization. AB - A premenopausal 45-year-old woman underwent uterine artery embolization for suspected symptomatic leiomyomata. Fourteen months later, with renewed symptoms and a new pelvic mass, metastatic leiomyosarcoma was diagnosed. A lack of clinical response to a technically successful embolization should alert care providers that further evaluation and/or therapy is needed. PMID- 15547556 TI - Three-dimensional ultrasound to differentiate epigastric heteropagus conjoined twins from a TRAP sequence. AB - Twin reversed arterial perfusion sequence and epigastric heteropagus conjoined twins may appear similar antenatally. Three-dimensional ultrasound evaluated the relationship of a completely formed fetus and an adjacent second body consisting of a pelvis with 2 lower extremities, confirming the final diagnosis when two dimensional ultrasound was unsuccessful. Three-dimensional ultrasound is useful in diagnosing epigastric heteropagus conjoined twins. PMID- 15547557 TI - Teaching residents how to teach improves quality of clerkship. AB - OBJECTIVE: To evaluate the impact of a resident's teaching skills workshop on the ratings of the obstetrics and gynecology clerkship by third-year medical students. STUDY DESIGN: The 6-week obstetrics and gynecology clerkship at the University of Michigan is provided at 4 different sites including the University of Michigan Hospital. At the end of each rotation, medical students complete an evaluation form assessing various aspects of their learning experience, including the overall quality of clerkship. A workshop, "Teaching Residents How to Teach," was conducted at the University of Michigan Hospital, whereas the other 3 sites served as the control. Clerkship evaluations were compared before and after the workshop using Student t test. RESULTS: Students at the University of Michigan Hospital rated all items on the evaluation form higher after the teaching workshop. The overall quality of the clerkship at the University of Michigan Hospital improved ( P = .05), whereas the other sites remained stable. CONCLUSIONS: A "Teaching the Residents How to Teach" workshop improves the overall quality of the clerkship. PMID- 15547558 TI - Impact of work hour restrictions on resident case experience in an obstetrics and gynecology residency program. AB - OBJECTIVE: The purpose of this study was to evaluate senior resident case experience before and after enactment of work hour restrictions. STUDY DESIGN: Obstetrics and gynecology experience from 2 postgraduate year 4 classes were evaluated before and after adoption of work hour restrictions. Data were limited to experience obtained during the fourth year of residency. Data were analyzed with the 2-sample t test and Wilcoxon rank sum test, and adjusted for changes in institutional procedural volume. RESULTS: There were significant decreases in resident experience in total abdominal hysterectomy ( P = .018), procedures for genuine stress urinary incontinence ( P = .004), and hysteroscopy ( P = .006). Decreases were seen in resident experience in vaginal birth after cesarean section ( P = .011), primary cesarean section ( P = .31), and vacuum delivery ( P = .007), despite increase in institutional volume. CONCLUSION: Work hour restrictions have had impact on resident case experience in obstetrics and gynecology. Variance in institutional case numbers account for only some of these changes. PMID- 15547559 TI - Teaching students to question. AB - OBJECTIVE: The purpose of the study was to teach medical students the strategies of critical questioning to determine the effect on students' critical thinking skills, confidence in their ability to ask questions, and interaction between student and instructor. STUDY DESIGN: Workshops were developed to teach medical students how to systematically ask critical questions. Sixty-two consenting students in their third-year obstetrics and gynecology clerkship were divided according to alternate rotations to either attend the workshops (n = 28) or not (n = 41). RESULTS: Medical students who attended the workshops scored higher on the California Critical Thinking Skills Test mean total score (study group 25.1 [+/- 0.7 SEM] vs control group 22.9 [+/- 0.6 SEM], P = .028), subscales of inference (12.6 [+/- 0.3 SEM] vs 11.2 [+/- 0.3 SEM], P = .003), and of deductive reasoning (12.7 [+/- 0.4 SEM] vs 10.9 [+/- 0.3 SEM], P = .001). CONCLUSION: Teaching students to ask critical questions improves critical thinking as measured by the California Critical Thinking Skills Test. PMID- 15547560 TI - Identification of educational objectives for obstetrics and gynecology residents in the ambulatory setting. AB - OBJECTIVE: The primary objective of this study was to identify specific educational goals for obstetrics and gynecology residents in the ambulatory setting. STUDY DESIGN: A cross-sectional study of current practice patterns in primary care, benign gynecology, and office procedures was performed with mailings to local private practice obstetrician-gynecologists. Questions for the anonymous written survey were generated using the Council on Resident Education in Obstetrics and Gynecology educational objectives. Telephone interviews with staff from billing offices confirmed practitioner responses. RESULTS: Of 88 practitioners, 43 (49%) responded. Diagnoses made in the office correlated well with the topics considered important for resident knowledge. Most important primary care diagnoses were depression and abdominal pain; important gynecologic diagnoses were abnormal uterine bleeding, chronic pelvic pain, contraception, and vulvovaginal infection. CONCLUSION: This study offers a valid, practical foundation for developing a focused ambulatory resident education program based on current outpatient obstetrics-gynecology practice patterns. PMID- 15547561 TI - Evaluating medical student obstetrics and gynecology clerkship performance: which assessment tools are most reliable? AB - OBJECTIVE: This study was undertaken to investigate medical student performance on an obstetrics and gynecology (Ob-Gyn) core clerkship to determine the most discriminating measures of student performance. STUDY DESIGN: Four-hundred twenty one clinical clerks were assessed with the use of 46 performance indicators in 4 different categories. Final Scores were weighted: (1) Clinical Performance = 60%, (2) Formal Presentation = 10%, (3) Oral Examination = 10%, and (4) National Board of Medical Examiners (NBME) Subject Test score = 20%. RESULTS: A total of 17,023 scores were examined. Final Scores were normally distributed. Clinical Performance and Formal Presentation scores were highly skewed, explaining 35% and 5% of the Final Score variance, respectively. Oral Examination scores were slightly skewed, explaining 17% of the variance. The NBME Subject Test score was the only normally distributed performance indicator and, despite comprising only 20% of Final Scores, was the most highly correlated ( R = 0.86), explaining 74% of the variance. CONCLUSION: Despite their minor weighting in our grading system, NBME Subject Test scores were very highly correlated with overall clinical clerkship performance. By combining written Subject Test and Oral Examination scores, 91% of the variance in Final Scores can be explained. PMID- 15547562 TI - The obstetrics and gynecology clerkship: building a better model from past experience. AB - OBJECTIVE: The study was undertaken to identify factors contributing to the poor student perception of the obstetrics and gynecology clerkship. STUDY DESIGN: Third-year medical students at the University of Michigan complete an annual questionnaire about the overall quality of their clinical experiences. In addition, at the end of each rotation, the students complete an evaluation form assessing various aspects of their learning experience. We reviewed data collected from 2000 to 2002. We calculated effect sizes using mean scores and SDs to compare individual aspects of the learning environment across the clerkships. RESULTS: Twenty-six percent of the third-year medical students at the University of Michigan rated the strength of their obstetrics and gynecology experience as very or exceptionally strong. Compared with the other 6 clerkships, the rotation had one of the lowest ratings for overall quality. Aspects of the clerkship experience we identified as potentially explaining this overall poor rating include the clarity of the clerkship goals and objectives, clarity of expectations for student performance, accessibility of faculty, experiences in learning history-taking skills, experiences in learning physical examination skills, and student perception that they were treated in a respectful/professional manner. CONCLUSION: Aspects of the clerkship experience identified by this study as potentially explaining the low ratings of the obstetrics and gynecology rotation should be studied in greater detail. Addressing these factors will be critical for improving the overall student perception of the obstetrics and gynecology clerkship. PMID- 15547563 TI - The development of a computer-assisted curriculum in reproductive endocrinology and infertility for residents. AB - OBJECTIVE This study was undertaken to determine the features of an effective computer-based residency curriculum in reproductive endocrinology and infertility. STUDY DESIGN: A review of the literature to determine those features that have been used by training programs in medicine. Reproductive endocrinology and infertility division directors, current obstetrics and gynecology residents, and obstetrics and gynecology practitioners in the community were surveyed to evaluate critical subjects for study. RESULTS: Programs most successful in using computers in training health care providers use a case-based approach that prioritizes important subjects. Areas of greatest importance to the 3 groups surveyed include infertility, dysfunctional uterine bleeding, and management of the climacteric, and several other areas were also deemed critical. CONCLUSION: Benefits of computer-based learning include consistency, the ability to develop problem solving skills for life-long learning, and a self-paced approach, but its validity as a teaching tool will require rigorous appraisal. PMID- 15547564 TI - Acquiring laparoscopic skill proficiency: does orientation matter? AB - OBJECTIVE: This study was undertaken to determine whether side-on laparoscopic operating orientation alters time-to-skill mastery compared with head-on orientation. STUDY DESIGN: One hundred thirty-one medical students were randomly assigned by operating axis (camera position to operating field) and completed 10 attempts at each of 5 previously validated laparoscopic skills stations. Time-to completion was recorded for each attempt, generating an orientation and skill station learning curve. Statistical analysis was performed by using repeated measures analysis of variance and linear, polynomial, and logarithmic models with 95% CIs. RESULTS: Sixty-eight students were randomly assigned to head-on orientation and 63 to side-on orientation. Comparing median time-to-completion by station, head-on attempts were faster than side-on attempts for every station. Comparing learning curves by orientation, side-on learning curves were steeper than head-on learning curves for every station, except one. Asymptotes were reached in both strata by the tenth attempt. CONCLUSION: Greater initial disorientation is seen with side-on orientation compared with head-on orientation. This can be overcome with practice. PMID- 15547565 TI - Teaching contraception: an interactive presentation using managing contraception. AB - OBJECTIVE: Our purpose was to determine whether an interactive medical student contraceptive teaching session results in improved knowledge gain or satisfaction when compared with a standard, didactic lecture. STUDY DESIGN: An interactive lecture was compared with a standard lecture among third-year medical students at Oregon Health and Science University School of Medicine and Medical College of Georgia. Knowledge gain was assessed by comparing correct responses to a posttest with the students' own responses to the same questions given as a pretest before the lecture. Satisfaction with the lecture was assessed on a visual analog scale. Scores and satisfaction ratings were compared within and between lecture groups. RESULTS: A total of 150 students participated. There was a significant increase in test scores within both the interactive and standard groups ( P < .0001 for both groups). The change in scores did not differ significantly between the groups ( P = .087). Although overall satisfaction was significantly greater with the standard lecture ( P = .004), it was high with both lectures. CONCLUSION: Knowledge gain with the interactive lecture was equivalent to that with the standard lecture. Satisfaction with both learning formats was high. PMID- 15547566 TI - Defining fetal station. AB - OBJECTIVE: This study was undertaken to determine the definitions and beliefs regarding fetal station among pregnancy caregivers. STUDY DESIGN: Residents, nurses, and faculty at 5 teaching centers in Denver, Colo, were given surveys to determine what definitions were being used for fetal station and the perceived importance of these definitions. RESULTS: There were 243 responses from the 453 surveys. We found 4 definitions were in use: level of the presenting part in relationship to ischial spines in (1) centimeters or (2) thirds, and level of the biparietal diameter in relationship to the ischial spines in (3) centimeters or (4) thirds. Few caregivers were aware that other caregivers were using different definitions of fetal station. CONCLUSION: This lack of standardization may lead to errors in the care of laboring patients. PMID- 15547567 TI - Accuracy of simulated cervical dilation and effacement measurements among practitioners. AB - OBJECTIVE: To determine whether hard or soft cervical models produced more accurate results when examined by practitioners of various experience levels. STUDY DESIGN: Simulated hard and soft cervical models were placed in a chamber designed to simulate a realistic vaginal examination. These cervical dilation and effacement models ranged from 1 cm to 6 cm and from thick (4 cm) to complete (0.2 cm). Each examiner had 20 seconds to evaluate each of 12 models of varying dilation and effacement using the soft and then the hard models. Models of the same dilation and effacement were presented to each practitioner in the same sequence. Physicians, nurses, and residents did the cervical evaluations. RESULTS: Of 360 recorded dilation measurements, only 19% were exactly correct using the soft models, whereas 54% were exact using the hard models. The percentages correct using hard cervix models progressively decreased with advancing cervical dilation. No consistent trend was found using the soft models. The respective results for correct effacement measurements were 49% soft versus 58% hard. CONCLUSIONS: Assessment of cervical dilation and effacement was consistently more accurate in the hard, compared with the soft, cervical model. The greatest deviation between the models was noted among the residents, suggesting that beginning practitioners may benefit from learning on firm models before progressing to more realistic soft models. However, it should also be noted that all practitioners might benefit from practice with soft models, especially to determine accurate dilation because scores in this category were consistently below what might be expected. PMID- 15547568 TI - Obstetrics and gynecology medical student outcomes: longitudinal multispecialty clerkship versus traditional block rotations. AB - OBJECTIVE: To evaluate outcomes of students in a longitudinal clerkship. STUDY DESIGN: Clerkship grades, written examination scores, and clinical evaluations were compared between students participating in a longitudinal clerkship and students in block rotations. The percent of students pursuing an obstetrics and gynecology residency from each group was evaluated. RESULTS: There was no difference in clerkship grade (3.3 versus 3.5, P = .158). Longitudinal and block students received similar clinical evaluations with no significant difference in any category. The average written examination score was lower for students in the longitudinal clerkship (76.9% versus 80.0%, P = .008). A higher percent of students in the longitudinal program chose an obstetrics and gynecology residency, 12.5% versus 6.17% (odds ratio = 2.17, 95% confidence interval 0.57 6.89, P = .18). CONCLUSIONS: Longitudinally trained students received similar clinical evaluations to their colleagues but may need assistance in acquiring the knowledge needed for the written examination. This program should be examined closely for factors influencing a higher percent of students to pursue a residency in obstetrics and gynecology. PMID- 15547569 TI - Resident job satisfaction: will 80 hours make a difference? AB - OBJECTIVE: This study was undertaken to assess job satisfaction and quality of life among obstetrics and gynecology residents before the 80-hour work week. STUDY DESIGN: We administered a job satisfaction survey to residents before July 1, 2003, assessing satisfaction with residency training, indicators of current quality of life, and predictions for the effect of reduced work hours. RESULTS: Residents were satisfied with training, with important outliers, including leisure time, ability to pursue educational reading, and surgical experience. We created job satisfaction facets that were generally reliable constructs and valid predictors for overall residency satisfaction. Residents predict more free time and a healthier lifestyle under the new requirements, but do not anticipate using additional time to study or teach. CONCLUSION: Job satisfaction facets for residents are proposed here and may be refined through further study. Lower scores for surgical experience are of concern in light of decreasing work hours. Educators must monitor self-directed learning efforts under new work hours. PMID- 15547570 TI - Comparing the effects of physical practice and mental imagery rehearsal on learning basic surgical skills by medical students. AB - OBJECTIVE: The purpose of this study was to test the effects of varying the amount of physical practice and mental imagery rehearsal on learning basic surgical procedures. STUDY DESIGN: Using a sample of 65 second-year medical students, 3 randomized groups received either: (1) 3 sessions of physical practice on suturing a pig's foot; (2) 2 sessions of physical practice and 1 session of mental imagery rehearsal; or (3) 1 session of physical practice and 2 sessions of imagery rehearsal. All participants then performed a surgery on a live rabbit in the operating theater of a veterinary college under approved conditions. Analysis of variance was applied to pre- and post-treatment ratings of surgical performance. RESULTS: Physical practice followed by mental imagery rehearsal was statistically equal to additional physical practice. CONCLUSION: Initial physical practice followed by mental imagery rehearsal may be a cost effective method of training medical students in learning basic surgical skills. PMID- 15547571 TI - Faculty and resident preference for two different forms of lecture evaluation. AB - OBJECTIVE: This study was undertaken to compare the use of 2 different lecture evaluation forms developed for obstetrics and gynecology residents to evaluate core curriculum lectures given by faculty. STUDY DESIGN: Content and delivery of faculty lectures were evaluated by residents using (1) a 10-question evaluation form that used a 5-point rating scale and (2) a checklist evaluation form that offered multiple options to improve lectures but provided no numerical score. Each form was used exclusively for 4 months. Faculty received feedback from both forms; then faculty and residents were surveyed regarding their preferences. RESULTS: A total of 384 rating scale and 398 checklist evaluation forms were completed during the study period. Residents preferred to complete the rating scale evaluation forms (67%), whereas faculty preferred feedback from the checklist evaluation forms (62%). Faculty were more likely to plan changes to their lecture format with feedback from the checklist evaluation forms (75% vs 25%, P = .031). CONCLUSION: Although devoid of structured positive feedback, the checklist lecture evaluation form was preferred by faculty. PMID- 15547572 TI - Assessing the impact of a comprehensive sexually transmitted disease curriculum on learning outcomes. AB - OBJECTIVE: Our purpose was to assess the impact of a curriculum designed to improve third-year medical students' knowledge of sexually transmitted diseases, measured by sexually transmitted disease-related items from the National Board of Medical Examiners subject examination and by a locally developed sexually transmitted disease test. STUDY DESIGN: All students (n = 108) were exposed to a new sexually transmitted disease curriculum: a 2-hour laboratory module, lectures, syllabus, and locally developed pretest/posttest with review of the test prior to taking the National Board of Medical Examiners subject examination. Students were randomized to a attend sexually transmitted disease clinic (n = 47) versus no sexually transmitted disease clinic (n = 61). RESULTS: Students performed equal to or better than the national average on 85% of the National Board of Medical Examiners sexually transmitted disease-related items after curriculum institution, compared with 56% of the test items prior ( P < .001). Magnitude of improvement was dependent on clerkship timing, with greater improvement in students taking the obstetrics-gynecology clerkship earlier in the third year. Mean postcurriculum test scores of sexually transmitted disease improved significantly ( P < .001), independent of clinic site and clerkship timing. CONCLUSION: The curriculum for sexually transmitted disease produced significant improvement in third-year medical students' knowledge of sexually transmitted disease. This might have an impact on future prevention and control of sexually transmitted diseases in communities in which these students practice. PMID- 15547573 TI - Labor and delivery nurses: a survey of attitudes toward third-year medical students and their education. AB - OBJECTIVE: Forty-six percent of third-year medical students at the University of Alberta rated labor and delivery nurses as a negative influence on their obstetrics and gynecology rotation. We hypothesized that the nurses would have mostly negative opinions toward students and their education. STUDY DESIGN: Labor and delivery nurses were surveyed with regard to their views on the following: (1) student learning objectives, (2) factors causing a patient to reject or accept a student, and (3) the role of the nurse in medical student education. RESULTS: Eighty-nine nurses ranked student tasks (objectives) as very appropriate, appropriate, neutral, inappropriate, and very inappropriate. History and physical examination, witnessing deliveries, assisting at deliveries, and following up women in labor were rated very appropriate or appropriate by more than 96% of nurses; 85% approved of students doing supervised deliveries. Nursing staff were equally divided in approving or disapproving of pelvic exams in labour and outpatient assessment by students. Artificial rupture of membranes, fetal scalp electrode application, and episiotomy repair were not approved of by more than 70% of nurses. The most important factors causing a patient to reject or accept a student were felt to be bedside manner and previous experience with a student, with least important being attractiveness, gender, and the nurse's opinion. Eighty-seven percent of nurses declared that one of their roles is to help students gain experience, but 71% said they would protect women from students with whom they were not comfortable. CONCLUSION: Labor and delivery nurses generally have a more positive attitude toward students and their learning than review of evaluations by the students would suggest. However, nurses have reservations about students performing technical procedures in the labor and delivery room. Creation of guidelines (objectives) with nursing input and better briefing of students with regard to nursing expectations may improve the student's experience. PMID- 15547574 TI - Is indomethacin harmful for fetus? How much? PMID- 15547575 TI - VEGF concentrations in pregnancy. PMID- 15547577 TI - Pudendal neuropathy is best determined by full neurophysiologic assessment. PMID- 15547578 TI - Failure of physiologic transformation of the spiral arteries of the uteroplacental circulation in patients with preterm labor and intact membranes. PMID- 15547582 TI - Can antenatal steroids be considered a fetal "stress" test in cases of severe placental insufficiency? PMID- 15547583 TI - WHI, Semmelweis, and the fallacy of "evidence-based medicine". PMID- 15547585 TI - Cost effective teamwork. PMID- 15547586 TI - Cause for concern? PMID- 15547587 TI - Dental manpower. PMID- 15547588 TI - Boosting numbers. PMID- 15547599 TI - Endodontics: Part 5. Basic instruments and materials for root canal treatment. AB - In this part the basic endodontic instruments necessary for effective root canal treatment are described. The properties of, and manufacturer's claims for, new instruments and techniques may be compared to these basic principles before they are purchased and introduced to clinical practice. Having the correct instruments for different clinical situations may make treatment both more efficient and more effective. PMID- 15547600 TI - Destructive membranous periodontal disease (ligneous periodontitis): a case report and 3 years follow-up. AB - Destructive membranous periodontal disease is a rare, destructive and poorly defined entity, which is the part of a systemic disease due to plasminogen deficiency and fibrin deposition. The disease is characterised by gingival enlargement and periodontal tissue destruction that leads to rapid tooth loss despite treatment attempts. Biopsy is essential to rule out other periodontal disease in the differential diagnosis. PMID- 15547608 TI - Epidemiological studies of tooth wear and dental erosion in 14-year old children in North West England. Part 2: The association of diet and habits. AB - OBJECTIVE: To determine the strength of association (expressed as Odds Ratios) of potential risk factors with erosion and tooth wear in 14-year-old schoolchildren. DESIGN: A random sample of 2,385 children were selected by a stratified two-stage technique based on schools and children. SETTING: Schools in NW England. METHODS: Tooth wear was assessed by one examiner on three surfaces of all 12 anterior teeth (labial, incisal and palatal) and the occlusal surface of all four first molars using a four-point scale. Enamel wear was scored 0, dentine exposure <1/3 scored 1, >1/3 scored 2 and secondary dentine or pulpal exposure, scored 3. A questionnaire enquired about general health, dental health, habits and the frequency of intake of a wide range of foods and drinks. RESULTS: The Odds Ratios for tooth wear on any surface for habits, reflux and certain foods were: bruxism, 1.10; stomach upset, 1.45; pickles 1.86; vinegar 1.36; salt and vinegar crisps 1.33; brown/other sauces 1.57. Similarly, the odds ratios for potentially erosive drinks were: fizzy drinks 1.32; sport drinks 1.58; herbal/lemon tea 3.97. The frequency of intake was bi-modal with 397 children drinking a can per day and 207 drinking two cans per day. A significant number drank acidic beverages at bedtime but this was not associated with dental erosion. CONCLUSION: Although odds ratios greater than unity indicate an association, this was not high for carbonated beverages and many other acidic foods or drinks. Examining at fourteen years may not be ideal, as the determinants of erosion/tooth wear have not acted for long, the indices do not discriminate sufficiently and proportionately few subjects have dentine exposed on smooth surfaces. PMID- 15547609 TI - The facilitating factors and barriers influencing change in dental practice in a sample of English general dental practitioners. AB - AIM: The objective of this study was to investigate the barriers and facilitators to change in dental practices among a sample of general dental practitioners (GDPs) from three regions of England. METHOD: In-depth face-to-face interviews with 60 GDPs were undertaken. The sample was selected from a group of 317 GDPs who had completed a questionnaire in the first phase of this study. The participants were selected to reflect diversity regarding the number and extent of self-reported changes reported in the questionnaire, and personal and practice characteristics. Of the 92 attempted contacts, 60 (65%) of the interviews were successfully completed. The interview schedule formed the basis of the interview. All the interviews and notes were transferred on to NUD*IST QSR version 4, a qualitative analysis package. RESULTS: No single factor was identified as being more important than another in facilitating change. The main facilitators for change were: financial factors, regular patient attendance, particularly a core patient group, staff loyalty, having regular staff meetings and open communication and having access to peer support. The main barriers to change were the reverse of the facilitators plus not having a financial stakehold in the practice. CONCLUSIONS: A range of factors were identified as influencing change in general dental practice. These include GDPs' attitudes and experience of change, patient factors, organisational issues, contact with peers and access to appropriate training courses. PMID- 15547610 TI - The role of dental therapists working in four personal dental service pilots: type of patients seen, work undertaken and cost-effectiveness within the context of the dental practice. AB - OBJECTIVES: To describe the type of patients seen and work undertaken by dental therapists employed in four personal dental service practices and to report on their cost-effectiveness within the context of the dental practice. METHOD: All members of the dental team used a standard day sheet to record all patient contacts and procedures undertaken in that session. Dental therapists recorded data for 30 consecutive sessions and dentists recorded information for 20 sessions. Items were recorded in sufficient detail to allow later matching with the GDS statement of remuneration and a calculation of the average gross fees and patient charges per session. RESULTS: The role of the dental therapist varied between the practices studied. In two practices the therapist saw a high proportion of child patients, and in one of these this was combined with providing care for a high number of adult patients who were exempt from patient charges. In the two practices where the dental team did not include a dental hygienist, the dental therapist had a relatively high workload providing dental hygiene care for adult patients. It appears that the gross fees and patient charges generated by the dental therapist in all four PDS practices fail to cover the cost of the salary of the dental therapist, dental nurse and associated overheads borne by the practice. PMID- 15547611 TI - The patient assessment questionnaire: a new instrument for evaluating the interpersonal skills of vocational dental practitioners. AB - This paper describes a pilot study aimed at evaluating a new instrument, the patient assessment questionnaire (PAQ), which uses patient ratings for the assessment of communication skills and professionalism in vocational practitioners (VDPs). The PAQ was developed as part of an assessment system designed to address all round competence. Acohort of 99 VDPs took part in the study. Questionnaires were distributed to consecutive patients in the general dental service at two time points in the training year. Data from the pilot study was analysed to determine whether the PAQ fulfilled the criteria for robust assessment. Results provide evidence of high levels of reliability, validity and feasibility of the PAQ instrument. All indications to date suggest that the PAQ will prove to be a valuable assessment tool. It is currently being evaluated as part of the system used to assess the all round competence of dental graduates undertaking vocational training in Scotland. PMID- 15547623 TI - Dribble's in... PMID- 15547625 TI - Linkage and mutational analysis of familial thyroid dysgenesis demonstrate genetic heterogeneity implicating novel genes. AB - The pathophysiology of thyroid dysgenesis (TD) is not elucidated yet in the majority of cases. The unexpected familial clustering of congenital hypothyroidism due to TD suggests a genetically determined disorder. Four genes have been hitherto involved in thyroid development, including migration and growth. Three of these encode transcription factors (the thyroid transcription factors 1 and 2 (TTF1 or NKX2.1 and TTF2 or FOXE1) and PAX8) while the other encodes the thyrotropin hormone receptor (TSHR). Some mutations have been reported in patients affected by thyroid defects, which supports the relevance of these four genes in TD. However, their involvement in the general TD population remains questionable. Therefore, to document their involvement, we performed a linkage analysis followed by mutational analysis in 19 multiplex TD families. The LOD score results failed to prove linkage between any of the four genes and the TD phenotype, whatever the postulated mode of inheritance. Manual extended haplotypes showed allele sharing among affected individuals of at least one of these four genes in the majority of families. Nevertheless, mutational analysis did not identify mutations in these cases, arguing in favor of identity by descent and not identity by state. Furthermore, as a main result of the present study, extended haplotypes confirmed by mutational analysis showed that the four genes were excluded in five out of the 19 investigated families, demonstrating the relevance of other genes. In conclusion, the present study demonstrates genetic heterogeneity in the TD disorder and suggests the involvement of novel genes. PMID- 15547626 TI - Supportive care needs of parents of children with cancer: transition from diagnosis to treatment. AB - PURPOSE/OBJECTIVES: To analyze research related to the pediatric oncology population supportive care needs from diagnosis to treatment. DATA SOURCES: Articles published from 1992-2002. DATA SYNTHESIS: 49 studies were included. All six categories of the Supportive Care Needs Framework were found, with most studies addressing one to three of the need categories. Informational (88%) and emotional (84%) needs were identified most frequently. CONCLUSIONS: No one study examined the entire range and types of supportive care needs from diagnosis to treatment. This knowledge is key to planning appropriate care and services. Future research should be directed at understanding the full constellation of needs encountered by parents during this time. Further refinement of the Supportive Care Needs Framework is required to fully define the categories of need. IMPLICATIONS FOR NURSING: Although more research is required, supportive care that focuses on informational and emotional support appears to be most important from diagnosis to treatment. Using a conceptual framework such as the Supportive Care Needs Framework provides a methodology for planning care based on needs. PMID- 15547627 TI - Herbs or natural products that protect against cancer growth part three of a four part series. AB - PURPOSE/OBJECTIVES: To provide evidence-based research information about 47 herbs and natural products that have the potential to protect against the development of cancer. DATA SOURCES: Natural Medicines Comprehensive Database and Lawrence Review of Natural Products-Monograph System. Information about these herbs has been found in evidence-based studies cited in the references. DATA SYNTHESIS: Early research shows that some herbs and natural products appear to have the potential to prevent cancer growth. CONCLUSIONS: This preliminary evidence may be useful to healthcare professionals or patients who are concerned about cancer. IMPLICATIONS FOR NURSING: Oncology nurses who receive this information can become resources for patients or other healthcare professionals. PMID- 15547629 TI - The watchful waiting management option for older men with prostate cancer: state of the science. AB - PURPOSE/OBJECTIVES: To summarize the recent literature and report the issues and controversies surrounding watchful waiting as a management option for prostate cancer. DATA SOURCES: All recent, published articles describing the experience, outcomes, and quality of life of men undergoing watchful waiting and the psychoeducational interventions tested in this population. DATA SYNTHESIS: The outcomes of men living with prostate cancer often do not vary greatly from men who are cured from the disease through radical prostatectomy or brachytherapy. Limited intervention studies have been aimed at improving these outcomes among those who have chosen watchful waiting. CONCLUSIONS: A paucity of information remains surrounding interventions to support men undergoing watchful waiting for prostate cancer. A consensus must be reached on who is most appropriate for watchful waiting. Watchful waiting does not mean doing nothing. Men who undergo watchful waiting should be assured that it is an active, deliberate process, not an opportunity to be overlooked by the healthcare system. IMPLICATIONS FOR NURSING: Future nursing care and research must concentrate on understanding the experience of men who are undergoing watchful waiting and interventions to improve outcomes in this population. PMID- 15547630 TI - Fatigue, weight gain, and altered sexuality in patients with breast cancer: exploration of a symptom cluster. AB - PURPOSE/OBJECTIVES: To identify the symptom cluster of fatigue, weight gain, and altered sexuality caused by treatment for breast cancer. DATA SOURCES: Published research and literature review articles. DATA SYNTHESIS: Fatigue, weight gain, and altered sexuality commonly occur after breast cancer chemotherapy. Each symptom has a significant impact on quality of life; however, viewing them as a symptom cluster magnifies their impact. CONCLUSIONS: These symptoms have yet to be studied as a cluster. Exercise appears to be an intervention common to each that may be effective in reducing the severity of these treatment side effects. IMPLICATIONS FOR NURSING: Nurses should view symptoms caused by breast cancer treatment holistically, keeping in mind that a reciprocal relationship often exists among symptoms. Identification of symptom clusters with empirically derived interventions may enhance quality of care and quality of life for patients. PMID- 15547631 TI - Determination of factors associated with hospitalization in breast cancer survivors. AB - PURPOSE/OBJECTIVES: To identify factors associated with hospitalization after diagnosis of breast cancer in working-age women. DESIGN: Descriptive, retrospective survey. SETTING: Caseload of a single medical oncologist affiliated with an urban, not-for-profit, academic medical center. SAMPLE: 123 consecutively evaluated women aged 21-65 years with breast cancer associated with projected survival greater than three years. METHODS: Data were collected from an electronic clinical file with demographic, diagnostic, and hormone replacement therapy (HRT) information. Four survey forms were mailed to subjects: (a) a form ascertaining personal demographics, health status, and healthcare utilization, (b) menopausal Symptom Rating Scale, (c) a hot flash diary, and (d) the Center for Epidemiologic Studies Depression Scale. MAIN RESEARCH VARIABLES: Menopausal symptoms, depression symptoms, age, time since diagnosis, and overnight hospitalization. FINDINGS: An increasing depression score and increasing menopausal symptoms score were found to be independent predictors of hospitalization controlling for age at diagnosis, disease stage, and time since diagnosis. Demographic variables, HRT use at or prior to diagnosis (a proxy measure of health status), current self-reported health status, and hot flashes were not associated with hospitalization. CONCLUSIONS: Psychological factors can be important significant predictors of hospitalization in survivors of breast cancer independent of disease stage. Further study should be undertaken to determine whether support services directed at identifying and treating those at risk for depression or menopausal symptoms may reduce the likelihood of potentially avoidable hospitalization. IMPLICATIONS FOR NURSING: The identification of those at high risk for hospitalization because of high levels of depressive or menopausal symptoms and prompt intervention offer the opportunity to improve the quality of life of breast cancer survivors and reduce the cost of health care for themselves, their families, and the healthcare system. PMID- 15547632 TI - Assessing coughing and wheezing in lung cancer: a pilot study. AB - PURPOSE/OBJECTIVES: To establish reliability and validity of two self-report questionnaires, the Lung Cancer Cough Questionnaire and the Lung Cancer Wheezing Questionnaire. DESIGN: Prospective, exploratory pilot study. SETTING: Clinical oncology settings in the southern United States. SAMPLE: 31 adult women with lung cancer. METHODS: Content validity of both questionnaires was assessed through a comprehensive literature review and an expert judge panel. Concurrent validity was established by Spearman rank correlation coefficients and Wil-coxon Rank Sum tests with items from other valid tools. Test-retest reliability was assessed by percent agreement, kappa, paired t tests, and correlations. Internal consistency was determined by Cronbach's alpha. MAIN RESEARCH VARIABLES: Cough, wheeze. FINDINGS: Cronbach's alpha showed excellent internal consistency and percent agreement, and kappa showed similarity of item responses across test-retest administrations. Nonsignificant paired t tests indicated similar mean scores, and significant test-retest correlations supported test-retest reliability. CONCLUSIONS: Preliminary testing indicates good reliability and validity for both questionnaires. Both instruments can identify people with problems of coughing and wheezing and have the potential for monitoring these symptoms over time and determining effectiveness of interventions. IMPLICATIONS FOR NURSING: Assessment of coughing and wheezing is an important component of monitoring respiratory symptoms of lung cancer. Both of these symptoms can be amenable to interventions. Further research is needed to confirm psychometrics and sensitivity of these tools. PMID- 15547634 TI - Radioimmunotherapy with tositumomab and iodine-131 tositumomab for low-grade non Hodgkin lymphoma: nursing implications. AB - PURPOSE/OBJECTIVES: To review radioimmunotherapy approaches for low-grade non Hodgkin lymphoma (NHL) with a focus on tositumomab and iodine-131 tositumomab (Bexxar, Corixa Corporation, Seattle, WA, and GlaxoSmithKline, Philadelphia, PA). Nursing implications for Bexxar therapy are reviewed, including radiation safety, patient education, and the management of therapy-related toxicities. DATA SOURCES: Journal articles, published research data, and clinical experience. DATA SYNTHESIS: The Bexxar treatment regimen (using an anti-CD20 antibody) consists of a dosimetric administration followed 7-14 days later by a patient-specific therapeutic administration. Infusion-related adverse events and myelosuppression are manageable. Patient and caregiver education regarding the benefits of radioimmunotherapy, treatment protocols, and radiation safety precautions is necessary. CONCLUSIONS: Bexxar therapy represents an important new treatment option for patients with low-grade NHL and can be administered on an outpatient basis. IMPLICATIONS FOR NURSING: Nurses play a vital role in the success of a Bexxar therapy program by providing patient and caregiver education, patient monitoring, and coordinating treatment schedules. PMID- 15547633 TI - Burden and depression among caregivers of patients with cancer at the end of life. AB - PURPOSE/OBJECTIVES: To examine the patient and family caregiver variables that predicted caregiver burden and depression for family caregivers of patients with cancer at the end of life. DESIGN: A prospective, longitudinal study was implemented with an inception cohort of patients and their family caregivers who were followed after the diagnosis and treatment of cancer. SETTING: Community oncology sites in the midwestern United States. SAMPLE: 152 family caregivers of patients with cancer who died during the course of the study. METHODS: Telephone interviews were conducted with patients at 6-8, 12-16, 24-30, and 52 weeks following diagnoses. In addition, patient medical records and state death certificates were reviewed. MAIN RESEARCH VARIABLES: Effect of caregiver age, gender, education, relationship to the patient, employment status, reports of patient symptoms, patient cancer type, stage of cancer, time from the patient's diagnosis to death, caregiver burden, and depression. FINDINGS: Caregivers aged 45-54 reported the highest levels of depressive symptoms, and caregivers aged 35 44 reported the strongest sense of abandonment. Caregivers who were the adult children of patients with cancer and those who were employed reported high levels of depressive symptoms. Feeling abandoned (a portion of caregiver burden) was more prevalent in female, nonspouse, and adult children caregivers, and adult children caregivers of patients with early-stage cancer and patients with multiple symptoms reported a high perception of disruption in their schedule because of providing care. Caregivers whose patients died early following diagnosis reported the highest depressive symptoms, burden, and impact on schedule. CONCLUSIONS: Caregivers reported levels of depression at thresholds for screening of clinical depression. The number of patient symptoms was related to levels of caregiver depressive symptoms. An association also was found between depression and employment status. Caregiver distress was not dependent on demands of care. IMPLICATIONS FOR NURSING: Very little research exists that prospectively analyzes family caregiver experiences of burden and depression when providing end of-life cancer care for a family member. Interventions aimed at decreasing caregiver depressive symptoms should be targeted to caregivers who are middle aged, adult children, and employed. Interventions aimed at decreasing the burden associated with feeling abandoned and having schedules disrupted while providing care should be targeted to caregivers who are female, nonspouse, and adult children, and caregivers of patients with early-stage cancer and multiple symptoms. PMID- 15547635 TI - Keep moving: patients with myeloma talk about exercise and fatigue. AB - PURPOSE/OBJECTIVES: To learn about the feelings, beliefs, and experiences of patients with multiple myeloma implementing an exercise program in the context of an aggressive tandem peripheral stem cell transplant protocol. RESEARCH APPROACH: Qualitative naturalistic (constructionist). SETTING: International referral center for the treatment of multiple myeloma. PARTICIPANTS: Purposive nonprobability sample of 12 men and 9 women with multiple myeloma aged 38-70 enrolled in the exercise arm of a randomized trial of epoetin alfa with or without exercise as an intervention to decrease cancer-related fatigue. METHODOLOGIC APPROACH: Verbatim transcripts of tape-recorded, in-depth interviews analyzed for thematic content using content analysis and constant comparison. FINDINGS: Themes included (a) belief systems, which encompassed participants' beliefs about exercise and epoetin alfa, philosophy, and self-concept, (b) social context, which included life before cancer, social environment, and social system congruence, and (c) intersection between belief systems and social context, which were participants' experience appraisals. All participants believed that exercise could be helpful and would recommend exercise to other patients with cancer in a similar situation. Most believed that lack of activity contributed to decreased energy. MAIN RESEARCH VARIABLE: Patients' experience of implementing exercise in the context of treatment for multiple myeloma. CONCLUSIONS: Complex interactions between participants' beliefs and social context and experience appraisal influenced their ability to adhere to an exercise program during aggressive treatment for multiple myeloma. INTERPRETATION: Through careful assessment, clinicians can capitalize on belief and social systems that support adherence to exercise as an intervention to ameliorate fatigue for patients with cancer who are undergoing prolonged aggressive treatment. PMID- 15547636 TI - The PRO-SELF pain control program improves patients' knowledge of cancer pain management. AB - PURPOSE/OBJECTIVES: To evaluate the effectiveness of a psychoeducational program (i.e., PRO-SELF Pain Control Program) compared to standard care in increasing patients' knowledge regarding cancer pain management. DESIGN: Randomized clinical trial. SETTING: Seven outpatient settings in northern California. SAMPLE: 174 outpatients with cancer and pain from bone metastasis. METHODS: Following randomization into either the PRO-SELF or standard care group, patients completed the Pain Experience Scale (PES) prior to and at the completion of the intervention. MAIN RESEARCH VARIABLES: Total and individual item scores on the PES. FINDINGS: Total PES knowledge scores increased significantly in the PRO-SELF group (21%) compared to the standard care group (0.5%). Significant improvements in knowledge scores for patients in the PRO-SELF group were found on five of the nine PES items when compared to baseline scores. CONCLUSIONS: The PRO-SELF Pain Control Program was an effective approach to increase patients' knowledge of cancer pain management. IMPLICATIONS FOR NURSING: The use of a structured paper and-pencil questionnaire, such as the PES, as part of a psychoeducational intervention provides an effective foundation for patient education in cancer pain management. Oncology nurses can use patients' responses to this type of questionnaire to individualize the teaching and to spend more time on the identified knowledge deficits. This individualized approach to education about pain management may save staff time and improve patient outcomes. PMID- 15547637 TI - Turbulent waiting: rural families experiencing chemotherapy-induced neutropenia. AB - PURPOSE/OBJECTIVES: To explore how rural families understand and manage the chemotherapy-induced neutropenia (CIN) experience. DESIGN: Qualitative, inductive approach using family interviews. SETTING: Family homes in a rural community in the midwestern United States. PARTICIPANTS: A convenience sample (7 families [21 people] who had a family member experiencing CIN) recruited from a regional cancer treatment center. METHODS: Semistructured family interviews that were recorded on audiotape occurred along with constant, comparative analysis over 12 months. An interdisciplinary research team analyzed the transcribed data using grounded theory methodology. MAIN RESEARCH VARIABLE: The family experience of CIN. FINDINGS: An overall family process of turbulent waiting with intensified connections was revealed. Families in the study experienced a sense of vulnerability in response to the diagnosis of CIN. Intensified connections existed within and beyond the families to nurses, physicians, and community members, emphasizing the value of relationships for rural families and highlighting trust in their care providers. Waiting for chemotherapy to resume created a sense of turbulence, an unsettling time described by families as "being on a roller coaster" or "dangling." To manage the period of waiting and protect the neutropenic patient, families developed family caring strategies, including inquiry, vigilance, and balancing. The process of turbulent waiting with intensified connections led families to a reframed family integrity that included an expanded capacity for caring and protecting. CONCLUSIONS: Rural families understand and manage CIN in a context of vulnerability. The threat posed by cancer is heightened by CIN. Family waiting is a rich, interactive process by which families reemphasize relationships to manage neutropenia and is a process that healthcare professionals should acknowledge. IMPLICATIONS FOR NURSING: Findings suggest the need for further investigation of family caring strategies and for the development of family-level assessment measures in the instance of CIN. Findings contribute to theory development regarding family cancer care and suggest a need to develop an intervention protocol constructed from the perspective of a family-professional partnership. PMID- 15547638 TI - Using narrative research to understand the quality of life of older women with breast cancer. AB - PURPOSE/OBJECTIVES: To elucidate some of the issues that affect the quality of life of older women (70 years of age and older) diagnosed with breast cancer. DESIGN: Descriptive design SETTING: A National Cancer Institute-designated site in the southeastern United States. SAMPLE: 12 women who were at least 70 years of age undergoing treatment (radiation, hormonal, or chemotherapy) for breast cancer. METHODS: Two to three interview encounters per participant, each lasting approximately one hour. FINDINGS: Eight major themes emerged: Importance of God, Positive Attitude, No Alteration in Lifestyle, Physician Trust, Caregiver to Others, Importance of Health, Importance of Family, and Alteration in Lifestyle. CONCLUSIONS: The eight major themes are similar in terminology but varied in individual meanings. IMPLICATIONS FOR NURSING: Nurses must determine whether older women with a diagnosis of breast cancer are also primary caregivers to other individuals. As the story-gatherers for the healthcare team, nurses can use the data derived from interviews to document patients' health histories and provide a therapeutic process of coping with illness. PMID- 15547639 TI - Psychological distress, fatigue, burden of care, and quality of life in primary caregivers of patients with breast cancer undergoing autologous bone marrow transplantation. AB - PURPOSE/OBJECTIVES: To determine the effects of sociodemographic variables, psychological distress, fatigue, and quality of life (QOL) on burden of care in primary caregivers of patients with breast cancer undergoing autologous bone marrow transplantation (BMT). DESIGN: Descriptive, correlational, predictive. SETTING: Urban National Cancer Institute-designated comprehensive cancer center in the eastern United States. SAMPLE: Convenience sample of 102 primary caregivers. METHODS: Participants completed the sociodemographic data form, Piper Fatigue Scale, Beck Depression Inventory, State-Trait Anxiety Inventory, QOL Index, and Measurement of Objective and Subjective Burden Scales prior to BMT during an oncology clinic visit. MAIN RESEARCH VARIABLES: Depression, anxiety, fatigue, QOL, and burden of care. FINDINGS: Mean fatigue scores were low. Primary caregivers experienced moderate state anxiety and low trait anxiety. Some experienced severe depression. The objective burden of care mean score was slightly higher than the subjective mean score. Mean QOL scores were low. All variables were significantly intercorrelated, except subjective burden and temporal and sensory fatigue. Significant correlations were found between age and subjective burden, and income with fatigue temporal subscale, and state and trait anxiety. Family subscale of QOL was a significant predictor of objective burden. Age and trait anxiety were significant predictors of subjective burden. CONCLUSIONS: Primary caregivers of patients with breast cancer scheduled for BMT experience fatigue, anxiety, burden of care, and low QOL. These caregivers may have tried to meet their needs and the patients' needs simultaneously. IMPLICATIONS FOR NURSING: Healthcare providers need to proactively assess primary caregivers of patients for fatigue, psychological distress, burden of care, and decreased QOL and provide nursing interventions tailored to individual needs. PMID- 15547640 TI - Intrinsic and extrinsic religiosity, spiritual well-being, and attitudes toward spiritual care: a comparison of israeli jewish oncology nurses' scores. AB - PURPOSE/OBJECTIVES: To examine the differences among secular, traditional, and religious Israeli oncology nurses' intrinsic religiosity, extrinsic religiosity, spiritual well-being, and attitudes toward spiritual care. DESIGN: A comparative, descriptive study. SAMPLE: 148 Israeli Jewish nurses drawn from the membership of the Israeli Oncology Nursing Association. METHODS: Nurses completed mailed questionnaires. The four scales used were intrinsic and extrinsic religiosity portions of the Revised Age Universal Intrinsic-Extrinsic Scale, the Spiritual Well-Being Scale, and the Spiritual Care Perspective Scale. FINDINGS: Secular, traditional, and religious Jewish respondents differed significantly in intrinsic religiosity, extrinsic religiosity, spiritual well-being, and religious well being. No significant differences were found in existential well-being and attitudes toward spiritual care. Although not significant, an interesting trend was that secular nurses demonstrated more positive attitudes toward spiritual care than religious nurses. CONCLUSIONS: Jewish nurses' religiosity, spiritual well-being, and perhaps their attitudes toward spiritual care may be influenced by whether they are secular, traditional, or religious nurses. IMPLICATIONS FOR NURSING: Israeli Jewish oncology nurses need self-awareness of their intrinsic religiosity, extrinsic religiosity, spiritual well-being, and attitudes toward spiritual care when they are administering holistic care to their patients. PMID- 15547641 TI - International Network for the Availability of Scientific Publications: facilitating scientific publishing in developing countries. PMID- 15547642 TI - Exploiting thiol modifications. PMID- 15547643 TI - Molecules that cause or prevent Parkinson's disease. PMID- 15547644 TI - Nicotine as therapy. PMID- 15547645 TI - Hemispheric asymmetries in biodiversity--a serious matter for ecology. PMID- 15547646 TI - Prevention and treatment of the metabolic syndrome. AB - The prevalence of the metabolic syndrome is increasing owing to lifestyle changes leading to obesity. This syndrome is a complex association of several interrelated abnormalities that increase the risk for cardiovascular disease and progression to diabetes mellitus (DM). Insulin resistance is the key factor for the clustering of risk factors characterizing the metabolic syndrome. The National Cholesterol Education Program (NCEP) Adult Treatment Panel (ATP) III defined the criteria for the diagnosis of the metabolic syndrome and established the basic principles for its management. According to these guidelines, treatment involves the improvement of the underlying insulin resistance through lifestyle modification (eg, weight reduction and increased physical activity) and possibly by drugs. The coexistent risk factors (mainly dyslipidemia and hypertension) should also be addressed. Since the main goal of lipid-lowering treatment is to achieve the NCEP low-density lipoprotein cholesterol (LDL-C) target, statins are a good option. However, fibrates (as monotherapy or in combination with statins) are useful for the treatment of the metabolic syndrome that is commonly associated with hypertriglyceridemia and decreased high-density lipoprotein cholesterol (HDL-C) levels. The blood pressure target is < 140/90 mm Hg. The effect on carbohydrate homeostasis should possibly be taken into account in selecting an antihypertensive drug. Patients with the metabolic syndrome commonly have other less well-defined metabolic abnormalities (eg, hyperuricemia and raised C-reactive protein levels) that may also be associated with an increased cardiovascular risk. It seems appropriate to manage these abnormalities. Drugs that beneficially affect carbohydrate metabolism and delay or even prevent the onset of DM (eg, thiazolidinediones or acarbose) could be useful in patients with the metabolic syndrome. Furthermore, among the more speculative benefits of treatment are improved liver function in nonalcoholic fatty liver disease and a reduction in the risk of acute gout. PMID- 15547647 TI - Association of traditional and nontraditional cardiovascular risk factors with coronary artery calcification. AB - Coronary artery calcification assessed by computed tomography is an emerging marker of coronary atherosclerosis. The authors examined the association of coronary calcium scores with traditional and nontraditional cardiovascular risk factors. In 1999-2000, they measured coronary artery calcium in 360 participants free of known coronary artery disease who had participated in 2 centers of the Atherosclerosis Risk in Communities (ARIC) Study. They related coronary calcium scores to risk factors measured in 1987-1989. Most traditional risk factors were associated with the coronary calcium score. For example, the multivariately adjusted odds ratio for an elevated score (> or = 100 versus < 100) was 3.5-fold greater per 10 years of age, 3.2-fold greater in men than in women, 3.1-fold greater with diabetes (statistically nonsignificant), and 1.4- to 1.7-fold greater per standard deviation greater increments of plasma cholesterol and pack years of cigarettes smoked. Carotid artery intima-media thickness also was positively associated with coronary calcification. In contrast, a wide variety of hemostatic and inflammatory markers and serum chemistry values were unrelated to calcium scores. These findings reaffirmed the established role of traditional risk factors in the etiology of coronary artery disease, as assessed by computed tomography, but did not identify any important nontraditional risk factors. PMID- 15547648 TI - Classic intermittent claudication is an uncommon manifestation of lower extremity peripheral arterial disease in hospitalized patients with coronary artery disease. AB - Synchronous peripheral arterial disease (PAD) and coronary artery disease (CAD) is common. Standardized questionnaires such as the Rose/WHO questionnaire and later the Edinburgh modification of this questionnaire were developed to screen for PAD. Little data are available on the sensitivity of these questionnaires in hospitalized patients with CAD. The aim of this study was to determine the accuracy of these questionnaires and the prevalence of classic intermittent claudication in hospitalized patients with CAD. Medically stable patients with CAD were invited to participate before hospital discharge. The patients answered both the Rose/WHO and Edinburgh modification claudication questionnaires and had an ankle-brachial index (ABI) measured. An ABI less than or equal to 0.9 was considered to be indicative of significant PAD. Patients who had undergone previous lower extremity revascularization for PAD and had a corrected ABI greater than 0.9 were excluded. Ninety-five patients (66 men) were recruited. By measuring the ABI, 35 patients (25 men) were found to have significant PAD. An additional 3 patients who had an ABI corrected by lower extremity revascularization were excluded from the analysis. The Rose/WHO questionnaire had a sensitivity, specificity, and overall accuracy (95% CI) of 14.3% (2.7-25.9%), 96.7% (92.1-100%), and 66.3% (56.8-75.8%), respectively. The Edinburgh modification of the Rose/WHO questionnaire had a sensitivity, specificity, and overall accuracy (95% CI) of 28.6% (13.6-43.5%), 90.0% (82.4-97.6%), 67.4% (57.9 76.8%), respectively. Despite the high incidence of synchronous PAD in hospitalized patients with CAD, traditional claudication questionnaires are insensitive to PAD detection. Classic claudication is an uncommon manifestation of PAD in hospitalized patients with CAD. PMID- 15547649 TI - The levels of soluble adhesion molecules in diabetic and nondiabetic patients with combined hyperlipoproteinemia and the effect of ciprofibrate therapy. AB - Cell adhesion molecules are thought to play a role in atherosclerosis. Several clinical trials have shown that fibrate treatment leads to a reduction in coronary events, although the mechanisms are not fully understood. Soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble E-selectin plasma concentrations were measured in 10 obese dyslipidemic men (group A), in 10 obese dyslipidemic type 2 diabetic men without coronary artery disease (CAD) (group B), and in 10 dyslipidemic type 2 diabetic men with angiographically documented CAD (group C) before and after 12 weeks of treatment with ciprofibrate. Compared with nondiabetic dyslipidemic men, diabetic patients with CAD or without documented CAD had significantly increased levels of sVCAM-1 (512 +/-39 versus 750 +/-139 ng/mL; p<0.0001 and 566 +/-78 ng/mL; p<0.01, respectively) and sE-selectin (54.8 +/-6.9 versus 65.9 +/-8.8 ng/mL; p<0.001 and 62.6 +/-9.4 ng/mL; p=0.056, respectively). The levels of sICAM 1 were similar in all 3 groups. Multivariate analyses showed that the higher sCAM levels in patients occurred independently of lipoprotein levels. Waist circumference as a marker of abdominal adiposity was the only independent predictor of elevated concentrations of all 3 cell adhesion molecules in multivariate analyses. sE-selectin was associated with HbA1C levels (p<0.01) in diabetic men at baseline. After 12 weeks of ciprofibrate therapy, sVCAM-1 levels were reduced by 13.5 +/-2.1%, sICAM-1 levels by 11.8 +/-2.2%, and sE-selectin levels by 17.1 +/-3.5% (p<0.01 for all) with the greatest sE-selectin reduction in the diabetic subgroups (p<0.001). There was no correlation between the lowering of soluble adhesion molecules and the magnitude of lipid-lowering effect. An increased level of circulating adhesion molecules may be a mechanism by which dyslipidemia and/or diabetes mellitus promotes atherogenesis, and treatment with ciprofibrate may alter vascular cell activation. PMID- 15547650 TI - Assessment of toe blood pressure is an effective screening method to identify diabetes patients with lower extremity arterial disease. AB - The authors evaluated a screening program for lower extremity arterial disease (LEAD) in diabetic patients and focused on the value of toe blood pressure assessment. They recruited 437 subjects, ages 30-70 years (134 healthy controls, 166 type 1 and 137 type 2 diabetic patients; control [Ctr], DM1, and DM2) with no previous history of LEAD. They were enrolled in a longitudinal study with a planned follow-up of 10 years. Patients were consecutively enrolled from outpatient diabetes units of 2 university hospitals. Subjects were screened with respect to peripheral circulation by use of established noninvasive techniques. These included arm, ankle (AP), and toe (TP) blood pressure measurements; evaluation of peripheral neuropathy; and a standardized physical examination. Results from the baseline examination are presented in this report. The number of patients who presented peripheral pressures or indices below normal (< mean -2 SD for controls) was higher among diabetic patients; 24% of DM1 and 31% of DM2, as compared to 6% of Ctr, had at least 1 lower limb with a low TP, AP, toe/arm index (TI), or ankle/arm index (AI), and these subjects were mainly identified by using the toe/arm index. TI was independently and negatively associated with fasting blood glucose in both patient groups, and with smoking, age, and diabetes duration in DM1. The mean AP was higher in the DM1 and DM2 groups compared to Ctr, whereas overall TP, TI, and AI were similar in the groups. It was also shown that abnormally low TI was significantly more common than low AI among diabetics (p<0.001), and this was true for TP vs AP as well (p<0.05). It is beneficial to include assessment of toe blood pressure and toe/arm blood pressure index to detect early LEAD in diabetic patients. Ankle blood pressure and indices alone are less efficient, owing probably to medial sclerosis in diabetic patients. Up to 30% of diabetic patients with no ischemic symptoms may have signs of impaired arterial circulation. PMID- 15547651 TI - Effect of chronic oral supplementation with vitamins on the endothelial function in chronic smokers. AB - Cigarette smoking has been associated with endothelial dysfunction including impaired endothelium-dependent flow-mediated dilation (FMD). In cigarette smokers, increased oxygen-derived free radicals have been suspected of being one of the major causes of endothelial dysfunction, owing possibly to the inactivation of nitric oxide by free radicals. Vitamins C and E are widely used antioxidant vitamins, which have also been reported to effectively improve the endothelial function in several conditions. To test the effect of moderate-term oral antioxidant vitamin supplementation on the endothelial function in smokers, the authors evaluated the combined effect of vitamins C and E, administered in normal dosages, on FMD in young male smokers. A prospective interventional study was performed. In 15 healthy male subjects (mean age, 24.4 +/-2.5 years old). They studied FMD in the brachial artery by using high-resolution ultrasound. The vascular effects of moderate-term oral supplementation with vitamin C (1.0 g/day) and vitamin E (500 mg/day) were determined during reactive hyperemia, which causes endothelium-dependent FMD. They performed a vascular function study 3 times including before vitamin supplement, after 25 days of vitamin supplement, and 4 weeks after the cessation of the vitamin supplement. The flow-mediated dilator response measurements were repeated twice a day before vitamin supplements, and the repeatability obtained from these measurements was found acceptable (variability of FMD <2%). The oral antioxidant vitamin supplement significantly restored FMD (3.8 +/-2.2% vs 5.9 +/-2.5%; p<0.05), however, this effect disappeared 4 weeks after the vitamin supplementations ended. The combined usual dosage of vitamins C and E supplements was found to improve the endothelial function in chronic smokers. PMID- 15547652 TI - The role of valvular and thoracic aortic calcifications in distinction between ischemic and nonischemic cardiomyopathy. AB - Determination of underlying etiology in patients with dilated and globally hypokinetic left ventricles may sometimes be difficult even after detailed history and complete clinical evaluation. Cardiac valvular and thoracic aortic calcifications have previously been reported to be used as a window to diffuse atherosclerosis of the vascular system. The authors prospectively examined the predictive value of mitral annular calcification (MAC), aortic valve calcification (AVC), and thoracic aortic calcification (TAC) in diagnosis of coronary artery disease as the underlying cause of diffuse left ventricular dilatation and systolic dysfunction. The study included 98 consecutive patients (male/female = 76/22, mean age = 58.9 +/- 10.7 years, range: 33 to 75 years) over the age of 30 years admitted to their clinics between October 1999 and December 2001 with signs and symptoms of congestive heart failure associated with documented cardiomegaly. Transthoracic echocardiography and coronary angiography were performed in all patients for the evaluation of valvular calcifications and coronary status. Although there was no significant difference between the groups with and without coronary artery stenosis (CAS), with regard to presence of MAC, patients with CAS tended to have MAC more frequently (12/61, 20% vs 4/37, 11%, p > 0.05). AVC and TAC were found to be significantly more frequent in patients with CAS compared to those without CAS (AVC, 35/61, 57% vs 4/37, 11%, p < 0.001 and TAC, 28/61, 46% vs 2/37, 5%, p < 0.001). While all 3 calcifications had sensitivity under 60%, and specificity and positive predictive value over 75% individually, the presence of any of them had a sensitivity of 80%, specificity of 86%, positive predictive value of 91%, and negative predictive value of 73%. Thus the presence of any of these calcifications distinguished patients with coronary artery disease with a sensitivity of 80% and specificity of 86%. The presence of aortic valvular valve and thoracic aortic calcifications seems to be associated with significant coronary arterial stenosis; however, with relatively low negative predictive values these cannot be used in clinical practice for diagnosis of underlying coronary artery disease in patients with dilated left ventricles and impaired systolic functions. PMID- 15547653 TI - Losartan reduces left ventricular hypertrophy proportionally to blood pressure reduction in hypertensives, but does not affect diastolic cardiac function. AB - In contrast to the well-recognized salutary effects of angiotensin-converting enzyme inhibition, the value of angiotensin II type I (ATl)-receptor blockade on left ventricular hypertrophy (LVH) is controversial. In addition, the data on the influence of this therapy on cardiac diastolic function are scarce. Thirty-nine patients with moderate primary hypertension, LVH, and normal systolic function received losartan, 50 to 100 mg daily. Transthoracic echocardiography was performed at baseline and after 6 months of treatment. Thirty-one patients completed and were included in the study (16 males, 61.1 +/- 1.0 years). The patients were divided into responders if mean blood pressure (BP) decreased > 5 mm Hg at the end of the study (20 patients) and non-responders (mean BP decrease < or = 5 mm Hg, 11 patients). The BP and the LVH were significantly reduced (systolic BP by 10.0%, diastolic BP 6.5%, mean BP 8.2%, left ventricular mass index [LVMI] 6.2%, interventricular septum 5.8%, posterior wall 3.0%) (p< or =0.02), attributed to the reduction of BP and LVH in responders; the LVH in non responders did not alter with treatment. A significant correlation was noted between changes in BP and LVMI (r=0.60, p<0.001). The systolic cardiac function remained normal. The Doppler parameters usually used to assess the diastolic function of the LV (early diastolic filling velocity [E wave], late diastolic filling velocity [A wave], ratio of E/A waves, isovolumic relaxation time), which were abnormal at baseline, did not change with treatment. The size of the left atrium increased (p<0.05) at the end of the study. In conclusion, a 6-month course with losartan decreased BP and LVH. However, the LVH regression was rather associated with the reduction of the hemodynamic stimulus per se, than any trophic effect of the drug in the myocardium. The diastolic cardiac function remained abnormal with treatment. PMID- 15547654 TI - Prediction of infarct-related coronary artery of patients with acute inferior myocardial infarction by a predischarge exercise test index. AB - The predictive accuracy of electrocardiographic markers in identifying the infarct-related artery of myocardial infarctions has been a subject of extensive investigation. The present study was designed to test whether the index L II/L III ratio adapted to exercise electrocardiograms could be utilized as a marker to distinguish right coronary and left circumflex arteries as culprit coronaries in acute inferior myocardial infarctions. For this purpose, 82 patients with a positive-symptom-limited and/or submaximal treadmill exercise test with modified Bruce protocol after an acute inferior myocardial infarction were studied. Those patients with ST segment elevation during the stress test were included in the study. ST segment index was defined as the ratio of exercise-induced ST elevation amplitude in L II/L III. Patients were classified as having an index > 1 (n=24) and < 1 (n=58), and the findings were compared with the findings on coronary angiography. The groups were comparable with respect to age, gender, peak exercise level, and double products achieved. Circumflex artery was the infarct related one in the majority (21/24; 88%) of patients with an index > 1, whereas most (51/58; 88%) patients with an index < 1 had the culprit lesion in their right coronary artery (p<0.001). The ratio of exercise-induced ST elevations in leads L II and L III has a significantly high ability to discriminate the infarct related coronary artery in patients with uncomplicated inferior myocardial infarction. Considering the prognostic importance of the type of coronary involvement, this index could be a part of predischarge evaluation in this patient group. PMID- 15547655 TI - Fibromuscular dysplasia of bilateral brachial arteries--a case report and literature review. AB - Fibromuscular dysplasia (FMD) is a noninflammatory, nonatherosclerotic disease of the vascular system, that can affect intima, media, or adventitia. The dysplasia in these layers leads to decrease in the lumen and increased turbulence leading to diminished flow through the affected portion. The treatment of FMD thus far has been either surgical and/or interventional. The authors describe a patient who presented with painful digits and who was found to have bilateral brachial artery FMD and was treated with balloon angioplasty with complete resolution of symptoms. PMID- 15547656 TI - Cardiac tamponade and superior vena cava syndrome in lung cancer--a case report. AB - A combination of pericardial effusion with cardiac tamponade and superior vena caval syndrome is an unusual first presentation of carcinoma of lung, although cardiac involvement is often a late finding in widespread malignancy. Clinical identification can be difficult antemortem. Accurate diagnosis and prompt intervention are necessary to prevent adverse outcomes. Decisions regarding treatment must take into account the clinical presentation and echocardiographic findings. Echocardiography-guided pericardiocentesis with catheter drainage and/or pericardial window is the primary treatment strategy of choice for most large or hemodynamically significant effusions. New cardiac symptoms or classic findings of cardiac tamponade should prompt aggressive investigation. We present a case of adenocarcinoma of the lung that initially presented as pericardial effusion with tamponade and superior vena cava syndrome. The patient had all the clinical features of tamponade such as pulsus paradoxus, tachycardia, elevated jugular venous pressure, hypotension, and electrical alternans on surface electrocardiography. The findings were confirmed on echocardiography and computed tomography of chest, both of which allowed for rapid confirmation of the presence of an effusion and compression of the superior vena cava. The existing literature on the subject is succinctly reviewed. PMID- 15547657 TI - Bacteremia caused by late-infected pacemaker lead--a case report. AB - A 73-year-old man with bradycardia and atrial fibrillation underwent implantation of a transvenous pacemaker system on the left anterior chest wall in 1995. Six years later, he was admitted for bacteremia from coagulase-negative Staphylococcus. Repeated treatment employing antibiotic therapy was ineffective. The infected electrode was removed under cardiopulmonary bypass. His electrode had become firmly encased with fibrous tissue within the right ventricle and atrium. It was removed under direct vision during complete cardiac arrest. The postoperative course was uneventful and there has been no recurrence after 1 year. PMID- 15547658 TI - Salmonella aortic aneurysm: suggestions for diagnosis and therapy based on personal experience--a case report. AB - Infectious aneurysm is a rare event, especially after the introduction of antibiotic therapy. However, its early detection is very important for timely treatment with antibiotics and surgical intervention. This pathology may generally be due to mycotic endocarditis or septic embolization, prevailing in the preantibiotic era, and to aortitis, whose incidence is actually increasing, mainly in subjects with preexisting large-vessel atherosclerosis and intimal defects. This clinical entity is usually defined as microbial arteritis and recognizes Salmonella spp as the microorganism most frequently isolated from blood or vascular tissue cultures. The authors present the case of a 56-year-old man with a history of hypertension that some weeks before admission manifested as hyperpyrexia and episodic lumbar pain, associated with hepatosplenomegaly and with a pulsing mass in the periumbilical region. Abdominal computed tomography (CT) scan documented a voluminous infrarenal aortic aneurysm with a markedly reduced and irregular vessel wall. The patient underwent surgical excision of the aneurysm, during which marked periaortic inflammation phenomena, complete absence of the posterior aortic wall for a length of 5-6 cm, and the exposure of the correspondent vertebral bodies were observed. Histopathologic examination of the aneurysmal tissue showed atheromatous and thrombotic aspects and confirmed strong signs of inflammation. This case may suggest that the occurrence of microbial aortitis, especially from Salmonella spp, should be taken into account in the presence of a septic status associated with back, abdominal, or thoracic pain. PMID- 15547659 TI - Double-orifice mitral valve associated with nonisolated left ventricular noncompaction--a case report. AB - Double-orifice mitral valve is a rare congenital anomaly. Although it is more frequently associated with other cardiac abnormalities, it may occur as an isolated lesion. There are 2 forms of myocardial noncompaction: isolated and nonisolated myocardial noncompaction. Nonisolated myocardial noncompactions are occasionally reported postnatally in association with congenital heart anomalies such as ventricular septal defect, pulmonic stenosis, and atrial septal defect. To our knowledge, this is the first case presentation reporting a double-orifice mitral valve associated with nonisolated myocardial noncompaction. PMID- 15547660 TI - Excision repair cross complementing-group 1: gene expression and platinum resistance. AB - Platinum compounds induce their cytotoxic effect by binding to a DNA molecule in the form of a platinum-DNA-adduct. Many previous studies have shown that the level of platinum-DNA-adduct correlats with response to platinum-based chemotherapy. Although the mechanism of platinum resistance in vivo is not clearly understood, laboratory studies on cancer cell lines suggest that nucleotide excision repair (NER) is the main mechanism responsible for this resistance by increased platinum-DNA-adduct removal. NER pathway is a network of many proteins gathered in a DNA-repair system. The excision repair cross complementing-group 1 (ERCC1) gene has the leading role in NER-pathway because of its damage recognition and excision ability. In this report we reviewed the pathway leading to ERCC1 gene transcription and translation in cancer cells when exposed to cisplatin. We summarized data from different cancer cell lines and human cancers showing that the high level of ERCC1-mRNA and/or ERCC1 protein is associated with resistance to platinum compounds with direct impact on cancer patient survival and finally we analyzed drugs interfering with ERCC1 gene expression and causing the reversal of the platinum resistance when given to cancer cells prior to platinum-based chemotherapy. PMID- 15547661 TI - ZNF268, a novel kruppel-like zinc finger protein, is implicated in early human liver development. AB - The advancement in gene knockout and transgenesis have brought about enormous improvement in our understanding of mouse embryogenesis in the past decade or so. On the other hand, relatively little is known about human embryogenesis due largely to the lack of easy access to human embryos and tissues for biomedical studies. We have previously isolated a novel zinc finger gene, ZNF268, from a 3 week-old human embryo cDNA library in an effort to identify genes important for human embryonic development. To investigate the potential involvement of ZNF268 in human embryogenesis, we report here the spatial and temporal regulation of its expression during development. Northern blot and Western blot analyses revealed that ZNF268 is expressed in early embryos, predominantly, if not exclusively, in fetal liver with little detectable expression in other fetal organs. Interestingly, unlike most zinc finger proteins, ZNF268 protein was found to be localized mainly in the cytoplasm of embryonic hepatocytes. This subcellular localization was substantiated by the localization of EGFP-ZNF268 fusion protein overexpressed in the transfected COS7 cells. These results suggest that ZNF268 plays a role in early human liver development most likely by functioning through a cytoplasmic mechanism. PMID- 15547662 TI - Two further AHO-like syndrome patients with deletion of glypican 1 gene region in 2q37.2-q37.3. AB - In this report, we describe two unrelated patients with mental retardation and brachydactyly E classified as patients suffering from Albright hereditary osteodystrophy-like (AHO-like) syndrome. Fluorescence in situ hybridization (FISH) analysis using 8 different subtelomeric probes in 2q36-37 proved that the patients had subtelomeric 2qter deletions of similar size. The recently proposed candidate gene glypican 1 (GPC1) is deleted in both reported patients. PMID- 15547663 TI - Investigation of MYCN status in neuroblastoma by fluorescence in situ hybridization. AB - Neuroblastoma, a form of neuroblastic tumor, is one of the most common neoplasms seen in early childhood. The diverse clinical behavior of this tumor is most probably explainable by the heterogeneity in the associated genetic changes. We investigated 12 neuroblastoma patients for MYCN amplification and chromosome 2 aneusomy by fluorescence in situ hybridization (FISH) and results were correlated with conventional cytogenetics. Samples from both primary tumor tissue and bone marrow metastasis were available in two cases. The copy number of MYCN oncogene paralleled that of chromosome 2 in 10 cases, whereas two cases (16.7%) showed numerous distinct signals within the nuclei of the tumor cells, consistent with MYCN amplification as double minute (dmin). The morphology of dmin in one case was of an extremely small type and might potentially be missed by conventional chromosome analysis. Discordant cytogenetics and FISH results was observed between primary tumor and metastasis disease in one case, with loss of chromosome 2 tetrasomic and pentasomic cells as well as over-representation of chromosome 2 disomic cells harboring MYCN amplification in bone marrow deposits. Our study reaffirmed the need for MYCN status to be determined in light of chromosome 2 copy number, as recommended by published guidelines. We also showed that genetic heterogeneity might occur between primary tumor and bone marrow metastasis. Finally, atypical dmin morphology when encountered would need confirmation by FISH study. PMID- 15547664 TI - Evaluation of sarcoglycans, vinculin-talin-integrin system and filamin2 in alpha- and gamma-sarcoglycanopathy: an immunohistochemical study. AB - The sarcoglycan subcomplex (SGC) is a well-known system of interaction between extracellular matrix and sarcolemma-associated cytoskeleton in skeletal and cardiac muscle. The SGC is included in the DGC made up of sarcoplasmic subcomplex and a dystroglycan subcomplex. Recent developments in molecular genetics have demonstrated that the mutation of each single sarcoglycan gene, causes a series of recessive autosomal muscular dystrophies, dystrophin-positive, called sarcoglycanopathies or limb girdle muscular dystrophies. Our recent studies have demonstrated that costameres are a proteic machinery made up of DGC and vinculin talin-integrin system, also revealing the colocalization of sarcoglycans and integrins in adult human skeletal muscle. These results may support the hypothesis of the existence of a bidirectional signalling between sarcoglycans and integrins in cultured L6 myocytes. The hypothesis of bidirectional signalling between sarcoglycans and integrins could be supported by the identification of a skeletal and cardiac muscle filamin2 as a gamma-sarcoglycan, delta-sarcoglycan and, hypothetically, beta1 integrin interacting protein. Our results, acquired with an immunofluorescence study on adult human skeletal muscle affected by LGMD type 2D and 2C, showed that in LGMD2D: a) alpha-sarcoglycan staining is severely reduced; b) the beta-gamma-delta-sarcoglycan subunit and all tested integrins staining are clearly detectable; c) filamin2 is normal and shows a costameric distribution. In LGMD2C: a) alpha-sarcoglycan staining is preserved; b) the beta gamma-delta-sarcoglycan subunit staining is severely reduced; c) the alpha7B integrin is slightly reduced and beta1D-integrin is severely reduced; d) filamin2 is severely reduced. Other tested proteins of the two systems show a normal staining pattern in both sarcoglycanopathies. Our study seems to confirm, for the first time on adult human skeletal muscle of subjects affected by LGMDs, the hypo theses of: a) the existence, in mouse myotubes in culture, of two distinct subunits in sarcoglycans subcomplex; b) the presence of a bidirectional signalling between sarcoglycans and integrins, previously demonstrated on rat cultured L6 myocytes; c) the interaction of FLN2 with both sarcoglycans and integrins. These results may stimulate the search of yet unidentified common interactors of both fiber-extracellular matrix interaction systems. PMID- 15547665 TI - Increased heme oxygenase-1 and decreased delta-aminolevulinate synthase expression in the liver of patients with acute liver failure. AB - Acute liver failure (ALF) remains a serious problem in critical care with a high rate of mortality. Although the pathophysiology of ALF has not been fully elucidated, oxidative stress has been in part implicated in its pathogenesis. Heme oxygenase-1 (HO-1) is known to be induced not only by its substrate, heme, but also by various oxidative stresses, and thought to play an important role in the protection of the host from oxidative tissue injuries. In the present study, we examined expression of HO-1 as well as the non-specific delta-aminolevulinate synthase (ALAS-N, or ALAS1), the rate-limiting enzyme in heme catabolism and biosynthesis, respectively, in the livers of patients with ALF. Compared with livers from control subjects who had various disorders, but normal hepatic function, HO-1 in the liver of ALF patients was highly up-regulated at both transcriptional and protein levels. Immunohistochemical studies demonstrated that HO-1 expression occurred predominantly in hepatocytes, but not in non-parenchymal cells. In contrast to HO-1, ALAS1 gene expression was markedly down-regulated in ALF patients compared with controls. These findings suggest that, in the liver of ALF patients, there may be an increase in free heme concentration which up regulates HO-1 gene expression, while down-regulating ALAS1 gene expression, resulting in markedly altered heme metabolism and liver function. PMID- 15547666 TI - Prevention of hydrogen peroxide-induced apoptosis of human peripheral T cells by a lysosomotropic iron chelator, ammonium chloride. AB - Human peripheral T cells are considered to be easily susceptible to oxidative stress because these cells lack peroxidase activity. Therefore, in a previous study, we investigated the site of ROS formation by utilizing Mito-Capture, H(2)DCFDA (succinimidyl ester of dichlorodihydrofluorescein diacetate), DAPI (4',6-diamidino-2-phenylindole), and LysoSensor. Our results showed that ROS formation was apparently diffusely distributed in T cells oxidatively stressed with 0.1 mM hydrogen peroxide. Moreover, lysosomal swelling and deformity, possibly revealing lysosomal membrane destabilization, were observed in these cells. Based on the above-mentioned results, we concluded that an apoptotic cascade involving early lysosomal membrane destabilization exists in the hydrogen peroxide-induced apoptosis of human peripheral T cells. Therefore, the possible involvement of lysosomal protease leakage caused by hydroxyl radical formation in lysosomes (possibly resulting in mitochondrial membrane dysfunction) is considered to play an important role in hydrogen peroxide-induced T cell apoptosis. Hydrogen peroxide-mediated destabilization of lysosomal membranes with release of hydrolytic enzymes such as many kinds of cathepsins into the cell cytoplasm can lead to a cascade eventuating in cell death. To assess the importance of the intralysosomal pool of redox-active iron, we examined the effect of blockade of lysosomal digestion by exposing T cells to the lysosomotropic alkalinizing agent ammonium chloride (NH(4)Cl). Preincubation of human peripheral T cells with 10 mM NH(4)Cl for 4 h dramatically decreased apoptotic death caused by subsequent exposure to hydrogen peroxide (H(2)O(2)), and lysosomes and mitochondria showed almost normally preserved appearance. Therefore, we concluded here that lysosomal protease leakage caused by hydrogen peroxide in T cells was prevented by preincubation with ammonium chloride (NH(4)Cl). PMID- 15547667 TI - Expression of collagen and fiber-associated proteins in human septal cartilage during in vitro dedifferentiation. AB - Chondrocytes surrounded by extracellular matrix are responsible for the maintenance of the cartilage as a functional entity. It is well accepted that chondrocytes cultivated for tissue engineering dedifferentiate in cell culture. We characterized the expression of different collagens and collagen related proteins in differentiated (primary) and cultured nasal chondrocytes by using microarray gene expression analysis and immunohistochemical staining. The genes for collagen subunits 1alpha1 (Col1alpha1) and 1alpha2 (Col1alpha2) were activated during a cell culture period of 5 and 20 days whereas Col2alpha1 could be detected both in differentiated and dedifferentiated chondrocytes. The long term cell culture revealed a late activation of the Col3alpha1, Col4alpha1 and Col11alpha1 genes as well as biglycan, fibromodulin and lumican. In addition, short- and long-term cell culture resulted in down-regulation of Col9alpha1, Col9alpha2, Col9alpha3, Col10alpha1, Col18alpha1, ColQ and chondroadherin. The decorin gene showed up-regulation in short-term cell culture, but down-regulation in long-term culture. Immunohistochemical staining of the different cell populations confirmed the mRNA data for collagen type 1, 2, 3, 4, 9alpha2, 9alpha3, 18 and decorin. Because of their up-regulation in cultured chrondrocytes the collagen types 1, 3, 4 and 11 as well as biglycan, fibromodulin and lumican may be markers for dedifferentiation. The collagen types 9, 18 and Q as well as decorin and chondro-adherin revealed down-regulation and, presumably, represent markers for the differentiation of chondrocytes. PMID- 15547668 TI - Pathophysiological role of human beta-defensins 2 in gastric mucosa. AB - Human beta-defensins (HBDs) recognized in the stomach include HBD1, which is the constitutional human beta-defensin (HBD), and HBD-2 and HBD-4, which are inducible HBDs. HBD-2 is an antimicrobial peptide that is involved in host defences against bacterial infections, such as Helicobacter pylori (H.pylori) in infection of the gastric mucosal epithelium. We examined the pathophysiological role of HBD-2, besides their roles as antimicrobial peptides. The materials used for the study consisted of gastric mucosal tissue specimens collected endoscopically from patients with conditions such as chronic gastritis associated with H. pylori infection, and gastric ulcers and gastritis due to non-steroidal anti-inflammatory drugs (NSAIDs) with or without H. pylori infection. We investigated the expression of HBD-2 and NF-kappaB by RT-PCR and immunoblotting, and the relation between the localization of HBD-2 and follicular dendritic cells (FDCs) by immunohistochemistry. Expression of HBD-2 was recognized in all the mucosal tissue specimens, irrespective of the presence or absence of H. pylori infection. All of the mucosal specimens expressing HBD-2 also revealed expression of NF-kappaB. In consecutive immunohisto-chemical staining, while expression of HBD-2 was observed in the gastric mucosal epithelium, FDCs were found to be localized in the lamina propria mucosae under the epithelial cell layer. These data suggested that in addition to being antimicrobial peptides, HBD-2 may also have a pathophysiological role as proinflammatory mediators, and that the HBD may act as proinflammatory mediators in concert with the dendritic cells (DC) by transmitting a signal from the mucosal surface to the lamina propria mucosae, which seems to be the original site of gastric mucosal damage. PMID- 15547669 TI - 2-O-methylisohemigossylic acid lactone, a sesquiterpene, isolated from roots of mokumen (Gossampinus malabarica) induces cell death and morphological change indicative of apoptotic chromatin condensation in human promyelotic leukemia HL 60 cells. AB - 2-O-methylisohemigossylic acid lactone, a sesquiterpene, was purified from roots of mokumen (Gossampinus malabarica) and identified by Mass, and (1)H- and (13) NMR. This sesquiterpene displayed strong growth inhibitory effect against human promyelotic leukemia HL-60 cells. Apoptotic morphological change of the nucleus, including chromatin condensation was induced in the HL-60 cells treated with the sesquiterpene. The fragmentation of DNA by the sesquiterpene to oligonucleosomal sized fragments, a characteristic of apoptosis, was observed to be dose- and time dependent in the HL-60 cells. Inhibitors of caspases suppressed the DNA fragmentation induced by the sesquiterpene. These findings suggest that growth inhibition by the sesquiterpene of HL-60 cells results from the induction of apoptosis by the sesqui-terpene, and that caspase cascade is involved in the induction of apoptosis by the compound in the HL-60 cells. PMID- 15547670 TI - Mesenchymal stem cells contribute to the renal repair of acute tubular epithelial injury. AB - Acute renal failure (ARF) is a common disease with high morbidity and mortality. Recovery from ARF is dependent on the replacement of necrotic tubular cells with functional tubular epithelium. Recent advancement in developmental biology led to the discovery of immature mesenchymal stem cells (MSCs) in bone marrow and several established organs and to the definition of their potential in the recovery from tissue injury. We investigated the effect of MSCs infusion on the recovery from ARF induced by intramuscle injection of glycerol in C57/BL6 mice. In this model, ARF is associated with an extensive necrosis of tubular epithelial cells due to myoglobin- and hemoglobin-induced injury. MSCs were obtained from bone marrow of transgenic mice expressing green fluorescent protein (GFP). MSC GFP-positive cells (MSC-GFP(+)) injected intravenously homed to the kidney of mice with glycerol-induced ARF but not to the kidney of normal mice. MSC-GFP(+) localized in the context of the tubular epithelial lining and expressed cytokeratin, indicating that MSCs engrafted in the damaged kidney, differentiated into tubular epithelial cells and promoted the recovery of morphological and functional alterations. Moreover, MSCs enhanced tubular proliferation as detected by the increased number of proliferating cell nuclear antigen (PCNA) positive cells. A significant contribution of the engrafted MSCs in the regeneration of tubular epithelial cells was shown by the presence of a consistent number of GFP(+) tubular cells 21 days after the induction of injury. In conclusion, these results indicated a tropism of MSCs for the injured kidney and a potential contribution of these cells to tubular regeneration and to the recovery from ARF. PMID- 15547671 TI - Biological fate of tissue-engineered porcine valvular conduits xenotransplanted in the sheep thoracic aorta. AB - The ideal prosthesis to replace the diseased human aortic valve is not yet available. We have previously shown that porcine acellular aortic-valve conduits, obtained by detergent-enzymatic method, display hemodynamic performances similar to those of their native counterparts. Hence, it seemed worthwhile to ascertain whether these tissue-engineered prostheses can be successfully xenotransplanted. Porcine acellular conduits, which immunocytochemistry demonstrated to lack MHC class I and II antigens, were implanted in the thoracic aorta of 9 sheep. Two animals died just after surgery, and the other 7 sheep were sacrificed 1 or 5 months after transplantation. A rather favorable outcome of the implant was observed in 4 sheep. In these animals, aortic valves remained pliable and coaptive, and the luminal surface of the conduits was endothelized just after one month from surgery. An intense inflammatory response was present at 1 month, and, although attennuated, it persisted for 5 months, located mainly between the tunica intima and media and at the border of the implant. Vimentin-positive and smooth muscle actin-positive myofibroblasts proliferated within tunica media and adventitia, and an obvious thickening of the tunica intima was also observed. Small vessels were seen in the adventitia, and elastic fibers were well-preserved in both the aorta wall and valve leaflets. In the cases of unfavorable outcome (3 of 7 survived sheep), implants were detached from the aorta recipient and surrounded by a connective mass that almost completely obstructed their lumen. These masses were composed of a fibromyxoid background where proliferating cells, resembling those occurring in human reactive myofibroblastic lesions (proliferative fascitis), were embedded. Collectively, these rather disappointing findings indicate that acellular valve conduits, obtained by the detergent enzymatic method, are presently not suitable for clinical applications because of the persistent inflammatory response, which conceivably triggers overgrowth mechanisms that lead to implant failure. PMID- 15547672 TI - Arg-Gly-Asp (RGD) peptide ameliorates carbon tetrachloride-induced liver fibrosis via inhibition of collagen production and acceleration of collagenase activity. AB - Liver cirrhosis is caused by a relative imbalance between synthesis and degradation of collagens. Arg-Gly-Asp (RGD) peptide is a major adhesive domain of several extracellular matrix (ECM) components, such as that involved in the binding of fibronectin to the alpha5beta1 integrin receptor. We previously reported that RGD peptide increased the expression of matrix metalloproteinase in hepatic stellate cells (HSCs) which play a major role in hepatic fibrosis. We evaluated whether RGD-peptides inhibit the progression of liver fibrosis in an animal model of carbon tetrachloride-induced hepatotoxicity. RGD peptide (GRGDS) (1 mg/kg body weight) was injected intraperitoneally (i.p.) 3 times a week for one month. The group treated with control peptide (GRGES) showed pathologically typical hepatic fibrosis, while the RGD-treated group showed minimal fibrotic changes. The liver contents of collagen and hydroxyproline in the RGD-treated group was significantly lower than that of the control group. Collagenase activity measured in liver homogenates was significantly higher in the treated group than in the control group. In an in vitro study using TWNT-4 cells derived from human HSCs, RGD peptide (100 mug/ml) reduced the expression of type I collagen and tissue inhibitor of matrix metalloproteinase-1, and increased that of matrix metalloproteinase-1. These results indicated that RGD peptides inhibited liver fibrosis associated with both decreased collagen production and increased collagenase acitivity, and suggested that RGD peptide might be useful for the therapy of hepatic fibrosis. PMID- 15547673 TI - Interleukin-5, interleukin-3 and granulocyte-macrophage colony-stimulating factor prime actin-polymerization in human eosinophils: a study with hypodense and normodense eosinophils from patients with atopic dermatitis. AB - Characteristic features of atopic diseases (AD) are immigration and local activation of eosinophils. Reorganization of the cytoskeleton modulates the function of leukocytes and is a prerequisite for the motility response. In this work, the regulation of actin polymerization has been investigated by flow cytometry using NBD-phallacidin and right angle light scatter measurements in purified eosinophils isolated from patients with atopic dermatitis and normal individuals. Stimulation of eosinophils with chemotaxins such as complement fragment C5a (C5a), CC chemokine RANTES/ CCL5 and platelet activating factor (PAF) induced a reversible polymerization of actin. Normodense eosinophils purified from patients with AD showed a decreased chemotaxin-induced actin response as compared to normodense eosinophils from healthy subjects and hypodense eosinophils from patients. Stimulation of eosinophils with Th2 cytokines such as interleukin-3 (IL-3), interleukin-5 (IL-5), granulocyte macrophage colony-stimulating factor (GM-CSF) did not exert a significant effect on actin polymerization. However, pretreatment with IL-3, IL-5 or GM-CSF potentiated the chemotaxin-induced actin polymerization and graded the differential responsiveness between normodense and hypodense eosinophils. We demonstrate a different actin responsiveness in eosinophils from atopic patients and healthy subjects which could be overcome by modulating effects of Th2 cytokines. PMID- 15547674 TI - Relationship between the -374T/A RAGE gene polymorphism and angiographic coronary artery disease. AB - The receptor for advanced glycation end products (RAGE) is thought to play a critical role in diabetic atherosclerosis. Accordingly, a functional -374T/A polymorphism in RAGE gene promoter has been associated with macrovascular complications in type 1 diabetic patients. However, the extent to which this common variant influences the risk of coronary artery disease (CAD) in the general population remains to be determined. We genotyped the -374T/A RAGE polymorphism in 259 non-diabetic individuals, of whom 175 had angiographically documented coronary artery disease (CAD patients) and 84 had normal coronary angiography (CAD-free control subjects). Homozygosity for the -374A allele was found in 9.7% of the CAD patients versus 22.6% of the CAD-free subjects (p=0.005). By means of a multiple logistic regression analysis, the AA genotype of the -374T/A polymorphism was shown to be independently associated with a reduced risk of CAD (adjusted odds ratio 0.33, 95% CI 0.15 to 0.73; p=0.006). Our observations suggest that the -374T/A polymorphism of the RAGE gene may reduce susceptibility to CAD, thus exerting a protective effect on coronary risk. Future pathophysiological studies may be worthwhile to clarify the mechanisms behind this association. PMID- 15547675 TI - Molecular characterization of human gastric mucosa by HR-MAS magnetic resonance spectroscopy. AB - The present study was aimed at identifying the molecular profile characteristic of the healthy human gastric mucosa. Ex vivo HR-MAS magnetic resonance spectroscopy performed at 9.4 Tesla (400.13 MHz for (1)H) on gastric specimens collected during endoscopy, permits the identification of more than forty species giving a detailed picture of the biochemical pattern of the gastric tissues. These preliminary data will be used for a comparison with gastric preneoplastic and neoplastic situations. Moreover, the full knowledge of the biochemical pattern of the healthy gastric tissues is the necessary presupposition for the application of magnetic resonance spectroscopy directly in vivo. PMID- 15547676 TI - Determination of HIV-1 infectivity by lymphocytic cell lines with integrated luciferase gene. AB - We have established lymphocytic cell lines H9 and M8166 that contain integrated copy of luciferase gene under the control of human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR). While H9 is known to be non-permissive for or insensitive to some particular mutant strains of HIV/simian immunodeficiency virus (SIV), M8166 is one of the most susceptible lines to various HIV/SIVs. The luciferase gene driven by HIV-1 LTR was transfected into H9 and M8166 cells with the neo gene, and cell lines were selected by G418. The indicator cell lines thus obtained were designated H9/H1luc and M8166/H1luc, and monitored for their susceptibility to various HIV clones including in vitro-constructed mutants. Both cell lines, particularly M8166/H1luc, were found to be exquisitely sensitive to HIV-1 and HIV-2. Furthermore, they exhibited the response to infections by various viral clones exactly as expected from the characteristics of the original cell lines. These results indicated that our new indicator cell lines H9/H1luc and M8166/H1luc are eminently useful for a variety of molecular virological studies on HIV/SIV. PMID- 15547677 TI - Role of albumin in osteoblastic cells: enhancement of cell proliferation and suppression of alkaline phosphatase activity. AB - The role of albumin in osteoblastic cells was investigated. Osteoblastic MC3T3-E1 cells with subconfluent monolayers were cultured for 24 to 72 h in a medium without fetal bovine serum (FBS) in the presence or absence of albumin (0.25, 0.5, or 1.0 mg/ml of medium). The number of osteoblastic cells was significantly increased by culture for 24 to 72 h in the presence of albumin (1.0 mg/ml). The effect of albumin in increasing cell number was completely abolished in the presence of PD98049 (10(-7) M), staurosporine (10(-7) M), or dibucaine (10(-7) M), which is an inhibitor of various protein kinases. Also, the stimulating effect of albumin on cell proliferation was completely prevented in the presence of cycloheximide (10(-7) M), an inhibitor of protein synthesis, or 5,6-dichloro-1 beta-D-ribofuranosylbenzimidazole (DRB), an inhibitor of transcriptional activity. DNA content was significantly increased in the cells cultured with albumin (1.0 mg/ml) addition for 24 to 72 h. Alkaline phosphatase activity in the cells, which participates in osteoblastic calcification, was significantly decreased by the culture with albumin (0.25, 0.5, or 1.0 mg/ml) for 24 to 72 h. The expression of insulin-like growth factor-I (IGF-I) and transforming growth factor-beta1 mRNAs using transcription-polymerase chain reaction (RT-PCR) analysis in osteoblastic cells was not significantly altered by culture for 48 h with albumin (1.0 mg/ml). This study demonstrated that albumin had a role in the regulation of osteoblastic cell function. PMID- 15547678 TI - Adrenomedullin and endothelin-1 stimulate in vitro expansion of cord blood hematopoietic stem cells. AB - The improvement of techniques for in vitro expansion of cord blood (CB) hematopoietic stem cells (SCs) is, at present, one main task of tissue engineering. Hence, we investigated whether endothelin-1 (ET-1) and adrenomedullin (AM), two regulatory peptides exerting growth promoting action on several cell systems, favor the in vitro expansion of CB SCs in liquid culture. CB hematopoietic cell middle-term expansion was carried out in a stroma-free liquid culture medium in the presence of ET-1, AM and three different cytokine combinations. After two weeks of incubation, aliquots of expanded-cell suspension were seeded on semisolid medium and clonogenic tests were carried out by counting the number of colony forming units (CFUs) after 14 days of culture. Neither ET-1 nor AM (2.5 x 10(-8) M) were per se able to significantly increase the CFU number, but both peptides magnified the pro-expansive effects of some cytokine cocktails. In light of these findings, we conclude that ET-1 and AM are to be considered novel promising molecules that, in association with cytokines, can be utilized as pro-expansive factors of CB SCs in prevision of their clinical use in allogeneic transplantation. PMID- 15547679 TI - Generation and characterization of HIV-1 clones chimeric for subtypes B and C nef. AB - The impact of human immunodeficiency virus type 1 (HIV-1) Nef on viral infectivity was evaluated by characterization of chimeric clones. Chimera with respect to the nef gene were constructed between subtypes B and C, and monitored for their replication in human peripheral blood mononuclear cells. The parental clones used were pNL432 (subtype B) and pIndie-C1 (Indian subtype C), which show considerable sequence heterogeneity in nef and distinct viral growth phenotype. While an enhancing effect of Nef on viral infectivity was noted, no significant growth difference was observed between the parental and chimeric clones. The difference in growth potential of the two subtype clones was mainly ascribable to viral sequence(s) other than nef. Our results here clearly showed that HIV-1 Nef does not significantly affect the in vitro viral infectivity in natural target cells. PMID- 15547680 TI - Induction of apoptosis by dideoxypetrosynol A, a polyacetylene from the sponge Petrosia sp., in human skin melanoma cells. AB - Dideoxypetrosynol A, a polyacetylene from the sponge Petrosia sp., is known to exhibit significant selective cytotoxicity against several human tumor cell lines. In the present study, we investigated the possible mechanisms by which dideoxypetrosynol A exerts its anti-proliferative action in cultured human SK-MEL 2 skin melanoma cells. Exposure of SK-MEL-2 cells to dideoxypetrosynol A resulted in growth inhibition and induction of apoptosis in a dose-dependent manner as measured by MTT assay, fluorescent microscopy and flow cytometry analysis. The increase in apoptosis was associated with a dose-dependent up-regulation in proapoptotic Bax expression and down-regulation of anti-apoptotic Bcl-2. Apoptosis-inducing concentrations of dideoxypetrosynol A induced caspase-3 and caspase-9 activation accompanied by proteolytic degradation of poly(ADP-ribose) polymerase and selective down-regulation of cIAP-1. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of dideoxypetrosynol A. PMID- 15547681 TI - Truncated hepatitis C virus core protein encoded in hepatocellular carcinomas. AB - Studies have suggested that a truncated form of the hepatitis C virus (HCV) core protein can enter hepatocyte nuclei and might play a role in HCV-associated hepato-carcinogenesis. In the present study, the HCV core gene from hepatocellular carcinomas (HCC) and/or adjacent non-tumorous liver tissues from eight patients was amplified by nested RT-PCR and sequenced. Mutations in the HCV core gene that would encode a truncated core protein were found in 4 of the 8 patients. Since truncated core proteins have been shown to be capable of entry into the hepatocyte nucleus (unlike HCV itself, which is an exclusively cytoplasmic virus), the detection of mutated sequences encoding them in these four HCC patients suggests that these mutations may have played a role in the development of these HCCs. PMID- 15547682 TI - Pseudohypoaldosteronism type 1 and the genes encoding prostasin, alpha-spectrin, and Nedd4. AB - Pseudohypoaldosteronism type 1 (PHA1), a rare disorder of infancy, presents with potential life-threatening salt wasting and failure to thrive. Thus far, PHA1 has been attributed to mutations affecting the mineralocorticoid receptor or any of the three subunits assembling the amiloride-sensitive epithelial sodium channel (ENaC). However, a lot of patients with a phenotype resembling PHA1, show no defects in these proteins, making it likely that further genes are involved in the aetiology of this disease. Recent studies have elucidated additional participants (alpha-spectrin and members of the families of transmembrane serine proteases, ubiquitin-protein ligases, and serum- and glucocorticoid-regulated kinases, respectively) regulating and/or interacting in the complex pathway of sodium retention in the amiloride-sensitive distal nephron. This led us to investigate whether PHA1 can also be associated with mutations in some of these genes. Our data suggest that at least the prostasin gene might be excluded as a causative locus. PMID- 15547683 TI - Frequencies of two common mutations (c.35delG and c.167delT) of the connexin 26 gene in different populations of Hungary. AB - The most common form of non-syndromic autosomal recessive deafness (NSRD) is caused by mutations in the gene GJB2, encoding the protein connexin 26 (Cx26). The mutation c.35delG is found in 30-70% of Caucasian NSRD cases, and is abundant (allele frequency of 0.5-2%) in several European populations, while c.167delT is found in the Ashkenazi Jewish population with about 2% frequency. In the current study, using simple PCR-based tests we established an allele frequency of 0.6% in the Hungarian average, and 0.4% in the Romani (Gypsy) populations for the c.35delG mutation, and an allele frequency of 2.4% in the Ashkenazi population for the c.167delT mutation. Our results do not differ significantly from the published data for Caucasian and non-European Ashkenazi populations and they present figures for the Romani population for the first time. Both mutations may be significant causative factors among the NSRD cases of the respective populations in Central Europe. PMID- 15547684 TI - Identification and characterization of FBXL19 gene in silico. AB - CXXC1, CXXC2 (FBXL10), CXXC3 (MBD1), CXXC4 (IDAX), CXXC5, CXXC6, CXXC7 (MLL), CXXC8 (FBXL11), CXXC9 (DNMT1) and CXXC10 are CXXC family genes within the human genome. Recently, we identified and characterized CXXC5 and CXXC10 genes as the homologs of CXXC4, which is implicated in the WNT signaling pathway. Here, we identified human FBXL19 (CXXC11) gene by using bioinformatics. Complete coding sequence of FBXL19 cDNA was determined by assembling 10 exons within AC135048.2 genome sequence. NM_019085.1 cDNA was a 5'-truncated partial cDNA corresponding to nucleotide position 138-2025 of FBXL19 complete coding sequence. FBXL19-BCL7C locus at chromosome 16p11.2, FBXL10-RHOF-BCL7A locus at chromosome 12q24.31, and FBXL11-RHOD locus at chromosome 11q13.2 were paralogous regions within the human genome. FBXL19 gene was found to encode a 674-amino-acid FBXL19 protein. Human FBXL19 showed 97.5% total-amino-acid identity with mouse Fbxl19. FBXHA domain (codon 11-128 of FBXL19) and FBXHB domain (codon 404-674 of FBXL19) were identified as novel domains conserved among FBXL19, FBXL10 and FBXL11. CXXC domain was located within the FBXHA domain, and F-box domain was located within the FBXHB domain. FBXL19 consists of FBXHA and FBXHB domains, while FBXL10 and FBXL11 consist of Jumonji C (JmjC), FBXHA and FBXHB domains. This is the first report on human FBXL19 gene as well as FBXHA and FBXHB domains. PMID- 15547685 TI - Role of histone deacetylase inhibitors in the treatment of cancer (Review). AB - Acetylation and deacetylation of nucleosomal histones play an important role in the modulation of chromatin structure, chromatin function and in the regulation of gene expression. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) are two opposing classes of enzymes, which tightly control the equilibrium of histone acetylation. An imbalance in the equilibrium of histone acetylation has been associated with carcinogenesis and cancer progression. So far, a number of structurally distinct classes of compounds have been identified as HDAC inhibitors including the short-chain fatty acids, hydroxamates, cyclic tetrapeptides and benzamides. These compounds lead to an accumulation of acetylated histone proteins both in tumor cells and in normal tissues. HDAC inhibitors are able to activate differentiation, to arrest the cell cycle in G1 and/or G2, and to induce apoptosis in transformed or cancer cells. Attention is currently being drawn to molecular mechanisms involving histone deacetylases. An induction of p21(WAF/CIP1) and a suppression of angiogenic stimulating factors have been observed in tumor cells following exposure to HDAC inhibitors. In xenograft models, several HDAC inhibitors have demonstrated antitumor activity with only few side effects. Several clinical trials showed that HDAC inhibitors in well tolerated doses have significant antitumoral activities. A combination of HDAC inhibitors with differentiation-inducing agents and cytotoxic drugs is an innovative therapeutic strategy that carries the potential for significant improvements in the treatment of cancer. PMID- 15547686 TI - Differential effects of Viscum album extract IscadorQu on cell cycle progression and apoptosis in cancer cells. AB - Extracts from European mistletoe or Viscum album L. have been reported to exert cytotoxic and immunomodulatory effects in vitro and in vivo. The mechanism of this anti-tumoral activity is however, largely unknown. In this study we tested the hypothesis that IscadorQu, an aqueous fermented extract from the European mistletoe grown on oaks, induces tumor regression by cell cycle inhibition and/or interference with apoptotic signaling pathways in cancer cells. Also a possible effect on angiogenesis, which is a prerequisite for tumor growth in vivo, is studied in endothelial cell cultures. Furthermore, we examined which apoptotic signaling route is activated by staining cells for specific pro-apoptotic proteins. To characterize these properties, 6 different human cancer cell lines, one epidermis derived cell line and 2 endothelial cell cultures were incubated with different concentrations of IscadorQu. Cell cycle kinetics parameters were measured by bromodeoxyuridine (BrdU) pulse labeling and tubulin staining. Apoptotic responses were detected by M30 CytoDeath or Annexin V/propidium iodide assays. Characterization of the apoptotic pathway was performed by staining cells for active caspase 3, active caspase 8, cytochrome C and chloromethyl-X-rosamine. The results of this study show that sensitivity to IscadorQu treatment varies strongly between different cell lines. In sensitive cell lines, including tumor and endothelial cell cultures, IscadorQu caused early cell cycle inhibition followed by apoptosis in a dose-dependent manner. Apoptosis was induced by activating the mitochondrial but not the death receptor-dependent pathway. PMID- 15547687 TI - Antimetastatic and anticancer activity of S-1, a new oral dihydropyrimidine dehydrogenase-inhibiting fluoropyrimidine, alone and in combination with paclitaxel in an orthotopically implanted human breast cancer model. AB - To evaluate the antitumor and antimetastatic efficacy of oral fluoropyrimidines, alone and combined with taxane on human breast cancer xenografts model, we developed a breast cancer model that spontaneously metastasizes to the lung by orthotopic implantation of MDA-MB-435S-HM tumors into the mammary fat pad (mfp) of SCID mice. The activity of the 5-fluorouracil (5-FU)-degrading enzyme dihydropyrimidine dehydrogenase (DPD) was significantly higher in the metastatic tumors than in the primary tumors. Based on this enzymatic characteristic of pulmonary metastases of breast cancer in regard to 5-FU metabolism, we investigated the antitumor activity of two types of oral 5-FU prodrugs, with and without paclitaxel, on both orthotopically implanted breast tumors and metastatic lung tumors in mice. The drugs and doses used were: S-1, a new oral DPD inhibiting fluoropyrimidine (DIF) 8.3 mg/kg/day, capecitabine 360 mg/kg/day as a non-DIF, and paclitaxel 50 mg/kg, all of which display minimal toxicity in mice. In the primary tumors, paclitaxel and S-1 displayed a significant antitumor activity, with 57 and 41%, respectively inhibition of tumor growth (p < 0.01), but capecitabine had no effect. When S-1 and paclitaxel were combined, they synergistically caused tumor regression (tumor growth inhibition ratio 94%, p < 0.01) in mice compared to capecitabine plus paclitaxel, without any toxicity. In the pulmonary metastasis model, paclitaxel, and both S-1 alone and combined with paclitaxel, but not capecitabine alone or combined with paclitaxel, diaplayed almost complete antimetastatic activity. These results strongly suggest that combination of S-1, as a DIF with taxanes will show a potent high antitumor and antimetastatic effect on refractory human breast cancers, especially those expressing strong DPD activity. PMID- 15547688 TI - Identification of antigenic epitopes recognized by Mac-2 binding protein-specific cytotoxic T lymphocytes in an HLA-A24 restricted manner. AB - We previously reported that 90K/Mac-2 binding protein (M2BP) is highly expressed in lung cancer and that M2BP-specific immunity was observed in many patients with lung cancer. These findings suggested the possibility of using M2BP as a target antigen in cancer immunotherapy. In this study, we selected 11 peptides derived from M2BP with an HLA-A24 binding motif and analyzed their ability to induce M2BP specific cytotoxic T lymphocytes (CTL). CTLs were generated with the M2BP-derived peptides from peripheral blood CD8-positive T lymphocytes of HLA-A24-positive healthy donors in multiple in vitro stimulations. Two CTLs, one induced with M2BP(241-250) (GYCASLFAIL) and the other with M2BP(568-576) (GFRTVIRPF), produced interferon-gamma in response to HLA-A24-positive TISI cells pulsed with the same peptide used for the in vitro stimulation. Although the CTLs induced with M2BP(241-250) reacted with both peptide-pulsed TISI cells and BT20 cells expressing both M2BP and HLA-A24, the CTLs induced with M2BP(568-576) did not react with BT20 cells. The cytokine production was blocked by antibodies against HLA class I in CTLs induced using M2BP(241-250), but not in CTLs induced using M2BP(568-576). These findings suggest that M2BP(241-250) is naturally processed from the native M2BP molecule in cancer cells and recognized by M2BP-specific CTLs in an HLA-A24 restriction. An M2BP-derived CTL epitope with an HLA-A24 binding motif was identified for the first time in this study, and it is expected to be useful as a target antigenic epitope in clinical immunotherapy for lung cancer. PMID- 15547689 TI - Wnt-signaling and apoptosis after neoadjuvant short-term radiotherapy for rectal cancer. AB - Recent surgical concepts for primary rectal cancer include the combination of surgery and short-term neoadjuvant radiotherapy (STNR). This is usually given in a dose of 25 Gy over five days in order to reduce local recurrence rates. Clinical studies have shown that local recurrence is found in some patients despite STNR. We identified molecular patterns of the Wnt- and apoptosis pathways as well as expression of junction-associated molecules in rectal cancer specimens of patients who received STNR and in those who did not. Expression patterns were examined by immunohistochemistry and molecular techniques such as LightCycler RT PCR and Western blot analysis in 25 sporadic rectal adenocarcinoma specimens derived from STNR-patients or non-pretreated donors, respectively. The molecular pattern in response to STNR was heterogeneous and was reflected by responders who show activation of apoptosis and cellular remodeling, whereas the group of non responders from STNR did not show such reaction and was very similar to untreated controls. Enhanced expression of beta-catenin was generally mediated by STNR, but exclusively in the responder group impaired expression of c-Myc and junction associated molecules as well as cleavage of poly-ADP-ribose polymerase and of the caspase substrate cytokeratin 19 were found. The molecular profile suggests that STNR interferes with Wnt-signaling and c-Myc expression. STNR in its present form is not suitable to fully complete the sequence of apoptosis in all rectal adenocarcinomas. PMID- 15547690 TI - HER2 analysis in breast cancer: reduced immunoreactivity in FISH non-informative cancer biopsies. AB - Due to the central role in predicting response to herceptin and possibly also other anticancer drugs, accurate and reproducible detection of the HER2 status is important. Fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) are the most commonly used methods for HER2 analysis. It is a disadvantage of FISH that a fraction of cases remain not interpretable probably due to suboptimal tissue handling before analysis. To investigate a possible influence of tissue damage on the results of HER2 IHC we compared the HER2 IHC results obtained in tumors with and without interpretable FISH in a breast cancer tissue microarray. The HER2 IHC results differed greatly between 1551 tumors with interpretable HER2 FISH signals and 405 breast cancers showing no FISH signals. FISH informative tumors had an IHC score of 3+ in 12.6%, 2+ in 3% and 1+ in 9.2% of cases. FISH non-informative tumors showed significantly lower IHC scores (p < 0.0001). They were IHC 3+ in 3.9%, 2+ in 3.7% and 1+ in 4.4% of cases. Overall, the data show that not only FISH but also IHC results are dependent on good tissue quality for successful analysis. Poor tissue quality can be easily identified in FISH analyses because of a lack of hybridization signals. Inappropriate tissue handling is more dangerous in IHC because an artificial lack of staining can be regarded as 'negative' result. PMID- 15547691 TI - Altered expression of cell cycle regulators Cyclin D1, p14, p16, CDK4 and Rb in nodular melanomas. AB - Cell cycle regulating proteins are important in tumour development. To investigate whether alterations in Cyclin D1, p14, CDK4 and Rb are associated with tumour cell proliferation, tumour progression and patient survival in malignant melanoma, we examined 202 vertical growth phase tumours and 68 corresponding metastases for expression of Cyclin D1, p14, CDK4 and Rb, and compared the results with Ki-67 expression, p16 and p53 expression, clinico pathological variables, and survival data. Nuclear staining of Cyclin D1 was strong in 35% of cases, and correlated with high levels of Rb (p=0.05), but not with survival or other markers tested. Strong staining of p14 was found in 63% of nodular melanomas and was associated with strong p53 expression (p=0.014), and with high levels of CDK4 (p<0.0001). Low p14 expression was associated with increased tumour thickness (p=0.008) and increasing level of invasion (p=0.020). Strong nuclear staining for CDK4 was found in 81% of cases and was associated with tumour thickness below the median value of 3.7 mm and improved survival (log rank test, p=0.024). Further, 56% of the tumours showed strong nuclear staining for Rb, and these cases were significantly associated with absent/low levels of p16 staining (p=0.030), high levels of p14 (p=0.010), as well as high Ki-67 expression (p=0.005). Our results seem to confirm that the p16-Rb pathway plays an important role in tumour progression and prognosis in vertical growth phase melanomas, whereas alterations in the p14-p53 pathway might be less important. PMID- 15547692 TI - Arrangement of expression and distribution of tight junction protein claudin-1 in cell dissociation of pancreatic cancer cells. AB - Mitogen-activated protein kinase kinase 2 (MEK2) was isolated previously as a potential factor related to cancer cell dissociation in highly (PC-1.0) and weakly (PC-1) invasive pancreatic cancer cells. On the other hand, changes of structure and function of tight junction (TJ) are reported to be correlated with carcinogenesis and tumor development. In this study, immunocytochemistry and Western blot analysis were performed in pancreatic cancer cells using anti claudin-1, MEK2 and phosphorylated MEK1/2 (p-MEK1/2) antibodies to reveal the correlation between TJ and cancer cell dissociation, as well as the involvement of MEK2 in regulation of TJ in cell dissociation of pancreatic cancer. After incubation with conditioned medium of PC-1.0 cells, plasma membrane distribution of claudin-1 was obviously disrupted, and expressions of MEK2 and p-MEK1/2, as well as dissociation of cell colonies, were significantly induced in PC-1 and CAPAN-2 cells. However, U0126 (a MEK1/2 inhibitor) treatment apparently induced the plasma membrane distribution of claudin-1 and aggregation of single cells in PC-1.0 and AsPC-1 cells, synchronously seriously suppressed MEK2 and p-MEK1/2 expression. Arrangement of expression and distribution of claudin-1 is closely related to cell dissociation status in pancreatic cancer cells through MEK2 activation. PMID- 15547693 TI - Sustained gene transfer and enhanced cell death following glucosylated-PEI mediated p53 gene transfer with photochemical internalisation in p53-mutated head and neck carcinoma cells. AB - p53 is frequently mutated in head-and-neck squamous cell carcinoma. Wild-type p53 gene transfer induces apoptosis in vitro and tumor regression in vivo and clinical investigations of p53 gene therapy have been reported, mostly using viral vectors. Non-viral vectors are increasingly being used as an alternative to viral vectors and photochemical internalisation (PCI) of non-viral vectors has been reported to yield high gene transfer efficiency. The p53-mutated status of FaDu human pharynx carcinoma cell line was first assessed by DNA sequencing and the cells were transfected using tetraglucosylated polyethylenimine (PEI-Glu4) in conjunction with photochemical internalisation (PCI). The green fluorescent protein (GFP) was used as a reporter for determination of the transgene expression kinetics with or without PCI. p53 gene transfer was performed in these optimised conditions, and subsequent induction of apoptosis was investigated by flow cytometric determination of the phosphatidylserine externalization. Long term cell death was assessed using colony forming assays. DNA sequencing in FaDu cells showed a G/T point mutation at codon 248 in exon 7 of p53 gene, resulting in an arginine-to-leucine substitution. As a consequence, P53 was shown to be expressed in >90% of untreated cells using immunocytochemistry. Using PEI-Glu4 as vector, PCI was found to significantly enhance GFP gene transfer whatever the formulation solution. Transfection efficiency was significantly increased with PCI. GFP expression kinetics (24-144 h) demonstrates that PCI induces sustained transgene expression with >10% of cells remaining transfected after 144 h. In such conditions, p53 gene transfer using PEI-Glu4 and PCI, resulted in spontaneous induction of apoptosis. As a consequence, long-term cell death was significantly enhanced after wt-p53 gene transfer when PCI was used, reaching up to 50% cell death. Wild-type p53 gene transfer using PEI-Glu4/DNA complexes and PCI, yields sustained transgene expression and induces cell death in p53-mutated FaDu cells. PMID- 15547694 TI - Identification and characterization of L985P, a CD20 related family member over expressed in small cell lung carcinoma. AB - We recently reported on the use of cDNA subtraction combined with microarray based expression analysis for identifying genes that are differentially over expressed in small cell lung carcinoma. One of the several hundred genes identified using this approach was termed L985P and its molecular characterization is described in this report. The differential over-expression of L985P mRNA in SCLC, as determined by microarray analysis, was confirmed by real time RT-PCR and Northern blot analysis. Immunohistochemical analyses show that L985P protein is highly expressed in SCLC with very restricted expression observed in normal lung, which was confined to the apical region of the ciliated bronchiolar epithelium. Flow cytometric and immunohistochemical analysis showed that L985P was localized to the cell surface. Sequence homology comparison indicated that L985P is identical to MS4A8B, a member of the recently described membrane-spanning 4-domain family, subfamily A (MS4A) gene family. The MS4A gene family currently consists of greater than 20 distinct human and mouse proteins that include CD20 and FcepsilonRIbeta. Both CD20 and FcepsilonRIbeta are involved in signaling events that regulate diverse cellular functions including cell growth regulation and differentiation. Collectively, the results presented herein demonstrate that L985P is differentially over-expressed in SCLC and may have potential clinical utility as an immunotherapeutic target for the treatment of SCLC. PMID- 15547695 TI - The clinical implication of 14-3-3 sigma expression in primary gastrointestinal malignancy. AB - 14-3-3 Sigma is a checkpoint control gene that promotes G2 arrest following DNA damage. The inactivation of the 14-3-3 sigma gene, primarily by methylation mediated silencing, has been reported in various human cancers. The loss of 14-3 3 sigma expression may contribute to malignant transformation by impairing the G2/M cell cycle checkpoint function, allowing an accumulation of genetic defects. In this report, we measured 14-3-3 sigma expression in 34 gastric and 35 colorectal cancers by using semi-quantitative reverse transcription-polymerase chain reaction and Western blot analysis. We also analyzed the association between 14-3-3 sigma expression and clinicopathological parameters including p53 status. Semi-quantitative reverse transcription-polymerase chain reaction and Western blot analysis showed that 14-3-3 sigma was significantly overexpressed in gastric and colorectal cancer tissues compared with normal ones (P<0.01). The immunoreactive 14-3-3 sigma protein was mainly detected in cytoplasm of cancer cells. Sigma overexpression tended to be associated with lymph node metastasis (P=0.08) in colorectal cancer. There was significant correlation between 14-3-3 sigma protein expression and the Ki-67 labeling index in gastric cancer (P=0.001). No significant association was observed between 14-3-3 sigma expression and p53 status. These results suggest that overexpressed 14-3-3 sigma in cancer cells might be induced by the p53 independent pathway, and that increased 14-3-3 sigma expression could contribute to cancer cell proliferation and the development and/or progression of human gastrointestinal cancers. PMID- 15547696 TI - Sensitization of multidrug resistant human ostesarcoma cells to Apo2 Ligand/TRAIL induced apoptosis by inhibition of the Akt/PKB kinase. AB - Chemotherapeutic agents have been used for the treatment of patients with osteosarcoma (OS). However, inherent or acquired resistance to these agents is a serious problem in the management of OS patients. The emergence of the multidrug resistance (MDR) phenotype in cancer cells is often associated with the overexpression of P-glycoprotein, encoded by the multidrug resistance gene MDR-1. The administration of some of the most common chemotherapeutic agents to these cells becomes ineffective because of their P-gp-driven efflux from the cell. Apo2L/TRAIL is a member of the tumor necrosis factor (TNF) family of cytokines that is considered to induce death of cancer cells but not normal cells. Its powerful apoptotic activity is mediated through its cell surface death domain containing receptors, TRAIL-R1/DR4 and TRAIL-R2/DR5, which in turn spread the signal in the cytosol through the activation of the caspase cascade. The Akt/PKB kinase is an important cell survival protein which is regulated by D3 phosphoinositides. High Akt expression and activity levels are well documented in many types of tumors, which very often show an altered PI3-K/Akt/PTEN pathway. In this study the U2OS human osteosarcoma cell line and its multidrug resistant (MDR) subline that overexpresses MDR-1 gene, MDR-U2OS, have been analyzed for their responsiveness to TRAIL. In conflict with the presence of active DR4 and DR5 receptors in both clones, U2OS cells exhibited only a low responsiveness to TRAIL, while the MDR-U2OS subline did exhibit a marked TRAIL sensitivity. An analysis of the post-receptor events showed that TRAIL responsiveness correlates with a reduced expression of endogenous Akt. In fact, expression in MDR-U2OS cells of a constitutively active Akt strongly decreased their sensitivity to TRAIL. The identification of Akt as a key modulator of TRAIL responsiveness could help to design TRAIL-based combinations for treatment of osteosarcoma. Moreover, the discovery that multidrug resistant osteosarcomas are highly sensitive to TRAIL-induced apoptosis indicates TRAIL as a new candidate for the treatment of multidrug resistant bone malignancies. PMID- 15547697 TI - Vasculogenic mimicry is associated with poor survival in patients with mesothelial sarcomas and alveolar rhabdomyosarcomas. AB - Increased vasculogenesis must occur for tumors to develop and be maintained. Normally, vascular networks are composed of tube structures lined with endothelial cells. However, the vascular networks that form around some highly aggressive cancers possess a distinct tubular structure, resulting from a process called vasculogenic mimicry (VM) that does not have endothelial cells. In these tubes, the tumor cells function as endothelial cells. VM has been found in several different types of cancers such as melanoma, breast cancer, prostate cancer, and ovarian cancer. We hypothesized that it also plays a role in the development and metastasis of sarcomas, which are typically aggressive tumors. We used immunohistochemical analyses and electron microscopy to identify VM channels in 81 synovial sarcomas (SSs), 37 mesothelial sarcomas (MSs), 69 alveolar rhabdomyosarcomas (ARs), and 190 melanomas, which were used as a comparison group. The presence of red blood cells in the vessels was also used as a criterion for VM. Because VM is generally believed to be associated with aggressive cancers, we tested whether the presence of VM channel correlated with patient survival. We detected VM channels in 11 of 81 SSs (13.6%), 10 of 37 MSs (27.0%), 13 of 69 ARs (18.8%), and 10 of 190 melanomas (5.3%). The VM channels were not distributed uniformly in the tumor tissues but appeared in patches. In addition, VM channels were most frequently observed in the boundary regions between the tumor and adjacent normal tissues. The tumor cells around the VM tubes frequently stained positive for collagen IV and CD31 and were also PAS positive. In contrast, tumors that lack VM channels generally also lack these markers. Our studies of the correlation of VM with patient survival also showed that VM correlated with shorter survival in patients with MS (P=0.03), AR (P=0.03), and melanoma (P=0.04), but not with SS (P=0.76). Our studies demonstrated that VM channels are a clinically important phenotype in sarcomas and melanomas. Our findings also suggested that a subpopulation of tumor cells possess features of both endothelial cells that line the vessels and mesenchymal cells that secrete the extracellular matrix required for the vascular infrastructure. PMID- 15547698 TI - Significance of PTEN alterations in endometrial carcinoma: a population-based study of mutations, promoter methylation and PTEN protein expression. AB - Mutations in the PTEN gene are frequent in endometrial carcinoma. PTEN methylation is an alternative mechanism of gene inactivation. To elucidate different mechanisms of PTEN gene inactivation, we have studied a population based series of endometrial carcinomas for PTEN mutations in relation to clinicopathologic characteristics, promoter methylation and protein expression. PTEN mutations were found in 54%, mainly in exons 5 and 8; with at least two different mutations in 21%. Presence of PTEN mutation was significantly correlated with young age, low FIGO-stage, endometrioid subtype, low grade, microsatellite instability and favourable prognosis. Previous studies of these tumours have observed PTEN methylation in 18% and low protein expression in 20%. Low expression of PTEN-antibody 6H2.1 was correlated with the presence of mutations in exon 8 among patients with 'two hits'; i.e. > or =2 mutations, or mutation(s) plus methylation (p=0.001). Number of PTEN hits was significantly associated with microsatellite instability, low hMLH1 expression and hMLH1 methylation. Thus, PTEN mutations are frequent in sporadic endometrial carcinoma and define a prognostically favourable subgroup, whereas the relationship with PTEN protein expression is complex. A pathway in endometrial carcinogenesis involving PTEN mutation and microsatellite instability is confirmed, and this study also indicates the importance of PTEN and hMLH1 methylation in this pathway. PMID- 15547699 TI - Expression of HLA class I antigen and proteasome subunits LMP-2 and LMP-10 in primary vs. metastatic breast carcinoma lesions. AB - Malignant transformation of breast epithelia is frequently associated with an altered expression of MHC products and of antigen processing molecular machinery. The consequent impairment of tumor immune recognition is thought to confer to tumor cells a selective advantage with respect to survival and metastatization. In order to understand if metastatic breast cancer lesions might be associated with a defective proteasome subunit expression that, in turn, might limit the peptide availability and prevent stable cell surface HLA class I-tumor antigen expression, we studied by immunostaining the expression of beta2-microglobulin, HLA class I antigens and proteasome subunits LMP-2 and LMP-10 in 35 matched primary and metastatic human breast carcinoma lesions. Overall, we found a downregulation of LMP-2 in 51.4% of the lesions, of LPM-10 in 45.7% of the lesions, of HLA class I heavy chain in 40.0% of the lesions, while beta2 microglobulin was downregulated in 25.7% of the lesions studied. In most primary and metastatic lesions the downmodulation of each antigen examined was coordinated. In the cases where a selective downmodulation of antigens was observed in the primary or in the metastatic lesion (with the exception of beta2 microglobulin), it was rather observed in the primary lesions. However, LMP-10 showed a significant selective downmodulation in the metastases as well. Antigen downmodulation does not appear therefore to represent a strategy for the primary tumor to metastasize successfully. PMID- 15547700 TI - Aurora-A/STK15/BTAK enhances chromosomal instability in bladder cancer cells. AB - Chromosomal aneuploidy is associated with invasive bladder cancer and one of the genes implicated in these changes is Aurora-A/STK15/BTAK, that is localized on chromosome 20q13 and encodes a centrosome-associated serine/threonine kinase. To better understand the association between Aurora-A/STK15 expression, tumor aneuploidy and clinical prognosis, we sought to determine whether overexpression of Aurora-A/STK15 in cultured urothelial cells facilitated chromosomal instability. Using immunofluorescence staining, Northern and Western blot analyses, we verified that overexpression of Aurora-A/STK15 in bladder tumor cell lines enhanced chromosomal instability. Additionally, we observed that some bladder tumor cell lines expressed more Aurora-A/STK15 than cultured normal urothelial cells and that Aurora-A/STK15 expression was higher in an immortalized E7 urothelial cell line having 20q amplification than in an E6 line lacking 20q amplification. These results were consistent with our observations of higher mRNA levels in some T3 invasive bladder tumors than in T1 superficial tumors and adjacent normal bladder tissue. Overall our results suggest that overexpression of Aurora-A/STK15 in bladder tumor cells contributes to tumor progression by promoting chromosomal instability leading to aneuploidy. PMID- 15547701 TI - Gene expression profiles of colorectal carcinoma in response to neo-adjuvant chemotherapy. AB - The combination of irinotecan and a fluoropyrimidine has been widely accepted as a treatment for advanced colorectal carcinoma. However, there have been no evaluable data on the feasibility of these combinations. To assess the significance of such combinations, we attempted to identify gene expression patterns in response to irinotecan and two different types of fluoropyrimidines. In 12 patients dispositioned to receive preoperative chemotherapy for colorectal carcinoma, pre-therapy tumor biopsies and final resected specimens were available for analysis. Patients were randomly assigned to receive one of the following four regimens: (I), oral doxifluridine; (II), intravenous infusion of 5-FU; (III), intravenous infusion of irinotecan; (IV), combination of doxifluridine and irinotecan (I+III). To identify genes whose expressions changed, we analyzed the gene expression profiles prior to and after these therapies using an oligonucleotide microarray consisting of 12,000 genes. Next, we focused on the genes that demonstrated similar kinetics of altered expression in all patients in each of the regimens. We identified two proto-oncogenes, nuclear receptor of T cells (NOT) and c-fos, that were up-regulated in doxifluridine- and irinotecan related regimens but unchanged in the 5-FU-related regimen. Moreover, group IV tumors showed the highest apoptotic rate and lowest proliferation activity following the combined chemotherapy. These results suggest that doxifluridine has a synergistic impact on the therapeutic effect of irinotecan by up-regulating proto-oncogenes such as NOT and c-fos, and thus justify the use of one of the irinotecan and fluoropyrimidine combinations. PMID- 15547702 TI - Comparative analysis of imbalances in genomic DNA and mRNA expression levels in chondrosarcoma-derived cell line FSCP-1. AB - Malignant cell transformation results from multiple biological alterations including chromosomal abnormalities, oncogene activation, loss of suppressor gene function and a imbalance in cell regulating processes. The aim of our study was to combine gene expression and genomic analysis to evaluate the cellular phenotype of a chondrosarcoma cell line, which is potentially a useful in vitro model system for physiological and/or neoplastic chondrocytes. cDNA-array, quantitative PCR and comparative genomic hybridization (CGH) technologies were used to analyze gene expression profiles of chondrosarcoma cell line FSCP-1 in correlation to changes of DNA copy number on corresponding chromosomal sections. Gene expression analysis revealed similarities, but also great differences in between the chondrosarcoma cell line and physiological chondrocytes. In particular the proliferative activity was up-regulated and molecules involved in matrix synthesis and turnover down-regulated. CGH analysis revealed a heterogeneous pattern of DNA gains or losses. The c-myc oncogene, located on 8q24.12-q24.13, was the only gene with a marked up-regulation located on a chromosome section with a gain of DNA copy number. The inability of the chondrosarcoma cell line FSCP-1 to maintain an adequate matrix turnover as well as a notable proliferative activity is similar to neoplastic chondrosarcoma in vivo. The limited correlation between the CGH analysis and the gene expression pattern supports the notion that also in neoplastic cells most genes are not primarily regulated by the gene dosage, but by cellular regulation pathways. However, genes such as c-myc might represent significant exceptions potentially relevant for the clinico-biological behavior of the neoplasms. PMID- 15547703 TI - Induction of p21 and nuclear accumulation of TAp73alpha and c-abl during apoptosis of cisplatin-treated primary pancreatic acinar cells. AB - To understand the role of endogenous p53 and related proteins in pancreatic injury responses, we established primary pancreatic acinar cultures from wild type and p53-deficient mice and investigated the relationship between apoptosis, proliferation and underlying molecular events in cells exposed to the DNA cross linking agent cisplatin. This treatment led to a time-dependent elevation in p53 levels, accompanied by phosphorylation at key serine residues. Despite this apparent activation of p53, acinar cells entered growth arrest unaffected by p53 deficiency. Moreover, p53-null cells exhibited only a temporal delay in engaging apoptosis, compared to wild-type counterparts. Whilst p53-proficient cells rapidly accumulated nuclear p21, the kinetics of p21 accumulation in p53-null cells were delayed, correlating with the execution of p53-independent apoptosis. During the course of treatment, c-abl and TAp73alpha, a p53 homologue, accumulated in acinar cell nuclei, irrespective of genotype, indicating that they are induced upon DNA damage and that they may act in parallel or in concert with p53 for the eradication of damaged acinar cells. We also report the nuclear accumulation of c-abl and TAp73alpha in cells, treated with the nuclear export inhibitor leptomycin B, suggesting that these proteins undergo constant nucleocytoplasmic shuttling in normal culture conditions, possibly reflecting a role for TAp73alpha-mediated transactivation or repression in the regulation of in vitro acinar cell growth. PMID- 15547704 TI - The role of reactive oxygen species in cisplatin-induced apoptosis in human malignant testicular germ cell lines. AB - Testicular germ cell tumours (TGCT) are the most common solid tumour among young males. Whereas in 1970s, only 5% of patients with a metastatic testicular tumours survived their disease, these days 80% of patients treated by modern cisdiamminedichloroplatinum (cisplatin, CDDP)-based chemotherapy are cured. Although data are accumulating on the effect of the mitogen-activated protein kinase (MAPK) family on the CDDP-induced apoptosis in tumour cells, the mechanisms by which CDDP initiates apoptosis in TGCT are not completely understood. Applying Western blot and phosphorylated kinase-specific ELISA analyses, flow cytometry, blocking experiments, and morphological methods we sought here to define the MAPK pathway(s) involved in the CDDP-induced apoptosis in the human TGCT cell line NCCIT. Our experiments showed that within hours of CDDP application only the extracellular signal-regulated kinase (ERK) was dually phosphorylated and caspase-3 became active. Functional assays using chemical inhibitors demonstrated that the phosphorylation of ERK was mediated by reactive oxygen species in an Raf-1-independent manner and required the activation of caspase-3. Thus, our data suggest that CDDP mediates its apoptosis-inducing effect on the human malignant testicular germ cells, at least partially, through activation of the MEK-ERK signaling pathway in a ROS-dependent, Raf-1-independent manner. PMID- 15547705 TI - Analysis of further disease progression in metastatic non-small cell lung cancer: implications for locoregional treatment. AB - To determine whether in the natural history of metastatic non-small cell lung cancer (NSCLC) a time interval exists when metastases are limited in number and/or destination organs. Thirty-eight stage IIIB (pleural effusion)/IV NSCLC patients were treated on a phase II trial of oxaliplatin and paclitaxel. Patients' charts were reviewed and all sites of disease at initial presentation and at subsequent follow-ups were recorded, including the number of organs involved and the number of individual metastatic sites. At presentation, 74% of patients had metastases confined to one or two organs (including the lung primary). Fifty percent had < or =3 metastatic sites in addition to the lung primary. At last follow-up, 17 patients developed new lesions, 14 in a new organ and 3 in a previously involved organ. Nineteen (50%) had stable (n=12) or progressive (n=7) disease in initially involved sites without developing any new metastatic tumors. Among the 17 patients who presented with < or =4 metastatic sites and no pleural effusion, 11 (65%) had stable or progressive disease in initially involved sites without developing new metastases. These results suggest that a subset of patients who present with metastatic NSCLC may not have widely disseminated disease and that some form of local treatment combined with systemic therapy might be beneficial in these patients. Our data support the feasibility of a clinical trial that incorporates local therapies to sterilize metastases in patients with NSCLC. PMID- 15547706 TI - Differential expression and splicing isoform analysis of human Tcf-4 transcription factor in brain tumors. AB - Tcf family transcription factors are the downstream effectors of the Wnt signal transduction pathway that regulates developmental and oncogenetic processes among species. The alternative splicing of consecutive exons in the 3' part of the Tcf 4 gene and an error-prone A9 repeat in exon 17 have recently been examined in several tumors. To further understand the roles played by differential expression and splicing isoforms of human Tcf-4 in brain tumorigenesis, the expression of the 3' part of the Tcf-4 gene (exons 10-17) was analyzed by RT-PCR, nested RT-PCR and DNA sequencing. The results showed that at least 13 of the Tcf-4 alternative splicing isoforms were found and most of them were overexpressed in various brain tumors. L3 isoform was particularly dominant in metastasis. Several novel splicing isoforms were identified. One that contains different combinations of exon 16 (10-11-12-16-17, S5) was found in normal brain and pituitary adenoma but not in astrocytoma, meningioma or metastasis, whereas the other that contains part of intron 16, designated exon-16', was found in metastasis. Overall 23% of sequencing analysis in brain tumors exhibited frameshift mutations in an A9 repeat region of exon 17. These mutants exhibited 2-base or 1-base deletion (A7, A8) and 1-base insertion (A10). Nonetheless, in vitro functional assay showed that these mutants did not affect the transactivity of Tcf-4 comparing to wild type Tcf-4. Collectively, our data suggest that a large number of alternative splicing isoforms may together with variable mutations of A9 repeat region maintain balanced pools of Tcf-4 isoforms during brain tumorigenesis. PMID- 15547707 TI - Reduced tumorigenicity of B16-F10 mouse melanoma cells transfected with mycobacterial antigen 85A. AB - Immunotherapy based on the administration of the mycobacterium bacillus Calmette Guerin has been successfully used in the treatment of in situ transitional cell bladder cancer, and may be applicable to the treatment of cutaneous malignant melanoma. Antigen 85A (Ag85A) and heat shock protein 65 kDa (hsp65) are major secreted proteins of Mycobacterium species and potent stimulators of cell mediated immunity. This study evaluated the ability of Ag85A and hsp65 gene transfection to limit tumor growth by B16-F10 mouse melanoma cells. Immunoblotting confirmed protein expression and secretion by B16-F10 cells that were transiently transfected with plasmid DNA containing the Ag85A or hsp65 gene. Groups of syngeneic C57BL/6 mice were injected subcutaneously with 1x10(5) untransfected B16-F10 cells or B16-F10 cells transiently transfected with either empty vector or vector containing the Ag85A or hsp65 gene. Ag85A-expressing B16 F10 cells exhibited a dramatic 76% reduction (p<0.05, Mann-Whitney U test) in tumor weight in comparison to empty vector controls at 14 days post-inoculation. In contrast, hsp65-transfected B16-F10 cells did not show any change in tumorigenicity. Decreased tumorigenicity by Ag85A-transfected B16-F10 cells was not due to a reduced ability of Ag85A-transfected B16-F10 cells to proliferate since both mock- and Ag85A-transfected B16-F10 cells showed increased in vitro proliferation in comparison to untransfected cells. Hematoxylin and eosin staining revealed that Ag85A-transfected B16-F10 tumors contained an inflammatory leukocyte infiltrate that was not present in hsp65-transfected tumors. Reduced tumor progression by Ag85A-transfected B16-F10 melanoma cells suggests that immunotherapy based on the transient induction of Ag85A expression may be an effective approach for the treatment of cutaneous malignant melanoma. PMID- 15547708 TI - The histone deacetylase inhibitor sodium butyrate induces breast cancer cell apoptosis through diverse cytotoxic actions including glutathione depletion and oxidative stress. AB - Sodium butyrate (NaBu), a potent histone deacetylase inhibitor, modulates the expression of a large number of genes. The purpose of this study was to determine whether this dietary agent could induce apoptosis in MCF-7 cells, a breast cancer cell line that lacks caspase-3 activity, and to identify the mechanisms that underlie NaBu toxicity in these cells. Cell viability assessed by the activity of mitochondrial succinate dehydrogenase (MTT assay) revealed a dose-dependent reduction of MCF-7 cellular growth in response to NaBu treatment. Restoring caspase-3 function by transfection did not modify NaBu toxicity in these cells. Following a 24-h exposure, NaBu-induced cell growth arrest in G2/M phase in a dose-dependent fashion in association with stable expression of CDC25A, a G1 specific regulator of the cell cycle. The anti-proliferative effects of NaBu were accompanied by diminished expression of p53. Similarly, mRNA encoding c-Myc, a well-known regulator of p53, was decreased in NaBu-treated cells, while p21(Waf1/Cip1) mRNA was increased. Furthermore, bax mRNA level was up-regulated whereas a decline in Bcl-2 both protein and mRNA levels were detected in NaBu treated cells. Apoptosis was observed following a treatment with 2 mM NaBu, reflected by Annexin-V staining and by the cleavage of poly(ADP-ribose) polymerase, whereas DNA laddering was absent. Apoptosis was associated with a pronounced depletion of intracellular glutathione levels. Finally, NaBu treatment significantly increased the activities of several antioxidant enzymes, including glutathione reductase, glutathione peroxidase, and catalase. Together, these data suggest that the pro-apoptotic effects of NaBu observed in MCF-7 cells are associated with oxidative stress. PMID- 15547709 TI - Evaluation of metastatic potential in prostate carcinoma: an in vivo model. AB - Prostate cancer is frequently associated with bone metastases, which are in fact the main cause of morbidity and mortality for this tumor. To better investigate this process, animal models of bone and bone marrow metastases need to be developed. However, experimental prostate cancer bone metastases are difficult to be obtained in vivo, and some typical clinical patterns remain irreproducible. In this study, we injected PC3 prostate cancer cells into the left cardiac ventricle of nude mice, thus reproducing the basic biological phenomenon of tumor cell spreading in the arterious blood stream, and compared the outcome with direct injection of cells in the bone marrow cavity of the tibia. Mice were monitored by X-ray analysis. After 40 days, 100% of intratibially-injected and 64% of heart injected mice revealed osteolytic lesions. The heart injection was then used to select PC3 cell subpopulations by serial inoculation and recovery from bone metastasis. One of the resulting cell populations, obtained by a second round of selection and denominated PCb2, showed a more invasive phenotype compared to parental PC3, both in vitro and in vivo. Although PCb2 cells had a growth rate comparable to that of PC3 cells, they generated a higher number of bone lesions in nude mice and crossed more easily the Matrigel barrier in vitro in the presence of several chemoattractants. This phenomenon was partially due to an increased capacity to adhere to laminin and to release MMP-2 at higher level relative to the original PC3 cells. This study demonstrates that heart injection of prostate cancer cells in nude mice may represent a good experimental model to investigate the pathophysiology of bone and bone marrow metastases in vivo. PMID- 15547710 TI - MAP kinase pathways involved in glioblastoma response to erucylphosphocholine. AB - Erucylphosphocholine (ErPC) is a promising antineoplastic drug for the treatment of malignant brain tumors. It exerts strong anticancer activity and induces apoptosis even in chemoresistant glioma cells. In the present study, A172 and U373MG glioma cells were treated with ErPC to explore the contribution of MAP kinase family members ERK, JNK and p38 kinase to ErPC-induced cell death. The exposure to ErPC led to activation of JNK and concurrent inhibition of ERK in both cell lines. Using specific MAP kinase inhibitors we confirmed that in U373MG cells ERK was blocked and JNK was activated upon ErPC treatment. Both effects were dependent on caspase activation. In A172 cells, ErPC treatment resulted in an activation of the JNK pathway, whereas the situation with respect to ERK signalling was more complex. We conclude that inhibition of the ERK pathway by ErPC may be related to antiproliferative effects, while activation of the JNK pathway may be responsible for its pro-apoptotic action. PMID- 15547711 TI - The BRAFT1796A transversion is a prevalent mutational event in human thyroid microcarcinoma. AB - Mutation in exon 15 of the BRAF gene is a characteristic feature of human thyroid papillary carcinoma (PTC). To determine the role of such mutation(s) in the neoplastic progression of thyroid papillary microcarcinoma (PMC), we analyzed 46 cases from 31 Russian and 15 Japanese patients with PMC. Mutated BRAF (the BRAFT1796A transversion in all cases) was detected in 13/46 (28.2%) of the tumors: 9/31 (29.0%) and 4/15 (26.6%) in Russian and Japanese individuals, respectively, displaying no signs of difference in the mutational rates in the PMCs from patients with diverse genetic background seen in PTCs. Occurrence of the BRAF mutation did not significantly correlate with the patients' gender, age at presentation, metastatic indices or with papillary, mixed papillary and follicular, and solid/trabecular PMC histotype. On the contrary, the tumors of follicular morphology significantly associated with the mutation-free genotype (P=0.018), and in the mixed-type tumors characterized by co-occurrence of well differentiated and less differentiated components, the BRAF mutational frequency was significantly elevated (P=0.020). The results indicate the BRAFT1796A mutation is prevalent in PMCs, and thus these tumors may have a spectrum of genetic events partly overlapping with that of PTCs. PMID- 15547712 TI - Efficacy of combination therapies of percutaneous or laparoscopic ethanol lipiodol injection and radiofrequency ablation. AB - Percutaneous radiofrequency ablation (RFA) is able to destroy hepatocellular carcinoma (HCC) in a few sessions without major complications. We have previously shown that not only the combined use of percutaneous ethanol injection and RFA (PEI-RFA) but also injection of mixture of ethanol and lipiodol (PELIT) was useful for the treatment of HCC. In the present study, we further developed the combined use of PELIT and RFA through percutaneous or laparoscopic approach (PELI RFA or LELI-RFA) and evaluated its usefulness. Nineteen nodules in 18 cases were treated with PELI-RFA or LELI-RFA. In the cases treated with LELI-RFA, no bleeding and no spilling milky fluid containing tumor cells were observed from the surface of ablated tumors. In the cases sufficiently treated with PELI-RFA or LELI-RFA, the mixture of ethanol and lipiodol was accumulated in the entire region of the tumor and low-density area was observed around the lipiodol deposit by computed tomography (CT). These delineations of coagulated area were helpful to evaluate the precise area of safety margin around the tumor treated with PELI RFA or LELI-RFA. Furthermore, the total volume of coagulated necrosis significantly and positively correlated with the product of energy requirement for ablation and the volume of ethanol injected by PELI-RFA or LELI-RFA. Among the cases treated with PELI-RFA or LELI-RFA, local recurrence emerged only in one case in whom enough safety margin could not be achieved by PELI-RFA. Therefore, it is critical to evaluate whether enough safety margin could be obtained with RFA therapy, and PELI-RFA and LELI-RFA are helpful in visualizing the safety margin area. PMID- 15547713 TI - Treatment of Jurkat acute T-lymphocytic leukemia cells by onconase (Ranpirnase) is accompanied by an altered nucleocytoplasmic distribution and reduced expression of transcription factor NF-kappaB. AB - Onconase (Ranpirnase), a novel ribonuclease isolated from Rana pipiens oocytes, was reported to suppress cancer cell growth in vitro, reduce tumor size in animals, and augment cytotoxicity of several chemotherapeutic agents. Since onconase is currently in phase III clinical trials tested in treatment of mesothelioma, much emphasis has been placed on the mechanism of its anti-tumor activity. Previous studies have shown that onconase-responsive cells become arrested at the G1/S checkpoint of the cell cycle and also undergo apoptosis. A proposed mechanism for these effects is that the enzymatic activity of onconase targets cellular RNAs, in particular tRNA, with an accompanying inhibition of protein synthesis. In the present study, we have investigated the time- and dose dependent effects of onconase on growth of Jurkat SN acute T-lymphocytic leukemia cells. Significant suppression of cell proliferation became evident after 72 and 96 h of treatment, and was most pronounced at the highest concentration (10 microg/ml; 8.3x10(-7) M) of onconase. This reduction of cell proliferation, however, was not accompanied by measurable changes in distribution of cells at different phases of the cell cycle, but was paralleled by the induction of apoptosis, as assayed by flow cytometry, and with a modest decrease in the expression of a cell cycle regulatory retinoblastoma protein (Rb). Further biochemical analysis revealed that growth suppression was closely coordinated with a down-regulation in the steady state and subcellular distribution of NF kappaB, a transcription factor known to be functionally associated with cell survival. The reduction in expression of NF-kappaB by onconase appeared to coincide or even precede growth suppression, suggesting a causal relationship. To further test the hypothesis that cellular localization and expression of NF kappaB may be critical to cellular response to onconase, we also studied the growth effects of onconase in Jurkat-BalphaM cells, which, unlike the parent SN T cells, contain a stably transfected dominant-negative IkappaB gene. Growth suppression by onconase in BalphaM cells was more pronounced and occurred earlier compared to SN cells, although still did not affect changes in cell cycle phase distribution. Contrary to expectation, however, diminution in NF-kappaB expression by onconase was even more pronounced in BalphaM cells, suggesting that this transcription factor, while presumably prevented from dissociation from its inhibitory protein IkappaB in these cells, is even more efficiently targeted for degradation by onconase. These results implicate NF-kappaB and its turnover as important determinants in the anti-proliferative/apoptotic effects of onconase in acute T-lymphocytic leukemia cells. PMID- 15547714 TI - Cancer cells as targets for lentivirus-mediated gene transfer and gene therapy. AB - Lentiviruses have been used as gene transfer vectors for almost 10 years and their utility has been demonstrated in a variety of different applications. However, their value in cancer gene therapy has not been studied thoroughly. Here we show that VSV-G pseudotyped HIV-1-based lentiviruses are efficient vectors for human tumor cells in vitro and in vivo. Lentiviral gene transfer efficiency was demonstrated by transducing 42 different cell lines, representing 10 different human tumor types. It was shown that most of the cell lines were good or excellent targets for lentiviral transduction, allowing 50-95% gene transfer efficiency. These results were comparable to those obtained with an E1/E3 deleted, serotype 5 adenovirus vector. Analysis of lentivirus vector structure revealed that virus particles devoid of HIV-1 accessory proteins appeared to be more efficient, but the presence of enhancing elements cPPT and WPRE did not play a major role in transduction efficiency to four different human tumor cell lines. However, their effect on the gene expression level in these cells was apparent. To examine the impact of lentiviral gene expression level on suicide gene therapy approach, human osteosarcoma cells were transduced with lentivirus- or adenovirus vectors carrying the fusion gene HSV-TK-GFP and exposed to ganciclovir. Cell viability analysis after the treatment revealed that both vector types induced similar level of cytotoxicity, suggesting that lentiviral expression of a suicide gene is adequate for tumor cell destruction. Finally, in vivo transduction studies with subcutaneous tumors showed that lentivirus vectors can yield similar gene transfer efficiency than adenovirus vector, despite three orders of magnitude lower titer of the lentiviral preparation. In conclusion, these data show that lentiviruses are efficient gene transfer vehicles for human tumor cells and justify their use in further preclinical cancer gene therapy studies. PMID- 15547715 TI - Mosaic subclinical melanoderma: an Achilles heel for UV-related epidermal carcinogenesis? AB - Cutaneous cancers are not uncommon on the face of elderly patients. Melanin should protect, at least in part, against the ultraviolet (UV)-induced neoplastic damage. However, the density in melanin chromatophores is heterogeneous in the epidermis of Caucasian adults. The computerized UV light-enhanced visualization (ULEV) method is a sensitive tool to assess non-invasively this mosaic pattern of intra-epidermal melanin load. In this study, the combination of ULEV pattern analysis and image analysis were performed involving four groups of phototype III Caucasian subjects. The first group was composed of 55 patients aged from 65 to 75 years who suffered from several malignancies of facial skin. The second control group of 55 patients who never had developed skin cancers were matched with the first group for age, sex and phototype. The third group was composed of 80 patients aged from 49 to 59 years who had developed a single basal cell carcinoma. The fourth group comprised 80 age, sex and phototype-matched healthy control subjects. Irrespective of the groups of subjects, a correlation was found between the pattern grading and the objectively determined relative area of subclinical melanoderma. Patients with multiple skin cancers differed from the other groups by the fact that a significantly higher proportion of them exhibited an extensive type of subclinical melanoderma. This feature was also seen in a minority of patients with a single basal cell carcinoma. The extensive subclinical melanoderma pattern is interpreted as a clue for risk, but not as a cause of UV-induced skin carcinogenesis. PMID- 15547716 TI - Preclinical testing of a peptide-based, HER2/neu vaccine for prostate cancer. AB - The HER2/neu protein is over-expressed in multiple epithelial tumors and the source of immunogenic peptides currently under investigation in vaccine trials in ovarian and breast cancers. We sought to define the correlation between HER2/neu expression and risk for prostate cancer recurrence and then determine the potential efficacy of anti-HER2/neu vaccination in prostate cancer patients at risk for recurrence. The risk for prostate-specific antigen (PSA) recurrence in 95 patients undergoing prostatectomy at the Walter Reed Army Medical Center (WRAMC) was calculated and correlated to HER2/neu expression, as determined by immunohistochemical staining. Peripheral blood lymphocytes (PBL) were then isolated from six consecutive human leukocyte antigen (HLA) A2+ patients with HER2/neu+ prostate tumors. These PBL were grown in parallel cultures and stimulated either with no peptide, HER2/neu E75 peptide, or control peptide. The cultures were compared for stimulated proliferation, induced peptide-specific cytotoxicity and tumor-specific cytotoxicity. When assessed by risk group, 69% of the high risk patients' tumors over-expressed HER2/neu compared to 47% of the intermediate risk group (p<0.05). Evaluation of the in vitro immune response of PBL isolated from six consecutive prostate cancer patients revealed a statistically significant increase in E75-stimulated lymphocytic proliferation. E75-stimulated lymphocytes demonstrated an E75-specific cytolytic response in 6/6 prostate cancer patients that increased with successive stimulations. Moreover, these E75-specific lymphocytes also demonstrated tumor-specific lysis against HER2/neu-expressing prostate cancer cell lines. The majority of prostate cancer patients at high risk for recurrence have HER2/neu expressing tumors. Hence, HER2/neu is a viable target for immunotherapeutics such as preventative immunization strategies with HER2/neu peptide vaccines. PMID- 15547717 TI - Adrenomedullin (AM) and AM receptor type 2 expression is up-regulated in prostate carcinomas (PC), and AM stimulates in vitro growth of a PC-derived cell line by enhancing proliferation and decreasing apoptosis rates. AB - Adrenomedullin (AM) is a hypotensive peptide, that acts via the calcitonin receptor-like receptor (CRLR), whose interaction with the subtypes 2 and 3 of a family of receptor activity-modifying proteins (RAMP) gives rise to two distinct AM receptors, named AM1 and AM2 receptors. AM derives from the post-translational proteolytic cleavage of pro(p)AM, the last step of which involves the conversion of the inactive AM to active AM by the peptidyl-glycine alpha-amidating monooxigenase (PAM). Compelling evidence suggests that AM, in addition to exerting its well-known regulatory action on blood pressure and water and electrolyte balance, also possesses a growth promoting effect in several normal and neoplastic tissues, including human prostate. Conventional reverse transcription (RT)-polymerase chain reaction (PCR) demonstrated the expression of pAM, PAM, CRLR and RAMP(1-3) mRNAs in both prostate hyperplasias (PH) and carcinomas (PC), and semiquantitative PCR showed that pAM, PAM and RAMP3 mRNA expression was higher in PCs than PHs. Radioimmunoassay measured higher concentrations of immunoreactive AM in PCs than PHs. The expression of pAM, CRLR and RAMP1,2 mRNAs was also detected in the PC-derived cell lines PC-3 and DU-145, RAMP3 expression being restricted to the latter line. AM did not affect the growth rate (duplication time) of PC-3 cells, but it did significantly increase that of DU-145 cells. The growth promoting effect of AM was found to ensue from both the rise in the proliferation rate and the lowering in the apoptosis rate of DU-145 cells. These effects of AM were counteracted by the AM receptor antagonists CGRP(8-37) and AM(22-52), the former antagonist, which is more selective for AM2 than AM1 receptors, being more effective than the latter one. Both antagonists were per se able to induce a slow, but significant decrease in the basal growth rate of DU-145 cells by inhibiting proliferation and enhancing apoptosis, again CGRP(8-37) being more effective than AM(22-52). Taken together, our findings allow us to suggest that: i) endogenous AM system plays an important autocrine-paracrine growth promoting action in the human prostate, being possibly involved in the development of the malignant phenotype of epithelial cells; and ii) the tumor promoting effect of AM in the human prostate is mainly mediated by the AM2 receptor (CRLR/RAMP3) subtype. PMID- 15547718 TI - Overexpression and amplification of STK15 in human gliomas. AB - The serine/threonine kinase 15 (STK15) at chromosome 20q13.2 is frequently shown to be amplified and overexpressed in several human cancers. STK15 has been reported to act as a cell cycle regulator and its overexpression induces centrosome amplification and aneuploidy. Recently we showed that STK15 even plays a role in human malignant brain tumours and we described an amplification of the gene in 31% of the investigated gliomas. In this study we scrutinized the correlation of increased STK15 on DNA and mRNA levels in gliomas of different histological grades. Southern blotting confirmed the amplification frequency of the STK15 gene, which had been previously detected by comparative PCR. In total, DNA gains were found in 26% of the investigated gliomas. Interestingly, we detected overexpression of STK15 mRNA in 60% of the analyzed brain tumours. The elevated expression does not strongly correlate with gains on DNA level, but all cases with an amplification of the STK15 gene display overexpression. Gains of the STK15 gene seem to occur irrespectively of the histological grades of the tumours, so that STK15 probably is not a progression associated factor. Amplification and overexpression of the kinase rather represent a primary alteration in human gliomas, which could play an important role as an early event in all glioma subtypes. PMID- 15547719 TI - Valproic acid and interferon-alpha synergistically inhibit neuroblastoma cell growth in vitro and in vivo. AB - Valproic acid (VPA) as a differentiation inducing anti-neoplastic substance is currently tested in solid tumour and leukaemia patients. Previously, we were able to show that the anti-cancer activity of VPA was synergistically increased by interferon-alpha (IFN-alpha) in Be(2)-C neuroblastoma (NB) cells. Now, we studied the effects of VPA in combination with IFN-alpha on two other NB cell lines. UKF NB-2 and UKF-NB-3 cell growth was synergistically inhibited by VPA and IFN-alpha. Cell cycle investigations revealed massive accumulation of cells in G0/G1-phase after a combined treatment with VPA and IFN-alpha. The VPA-induced accumulation of acetylated histones in NB cell nuclei that indicates inhibition of histone deacetylases was not further enhanced by the combination treatment with IFN alpha. Most strikingly, VPA plus IFN-alpha synergistically inhibited growth of UKF-NB-3 xenograft tumours in nude mice and induced complete cures in two out of six animals, while single treatment merely inhibited tumour growth. The results of this study together with our previous report strongly encourage the clinical evaluation of VPA and IFN-alpha for NB patients. PMID- 15547720 TI - Equiguard suppresses androgen-dependent LNCaP prostate cancer cell proliferation by targeting cell cycle control via down regulation of the retinoblastoma protein Rb and induction of apoptosis via the release of cytochrome c. AB - Lack of life-prolonging therapies has provided much of the impetus for seeking complementary and alternative management/treatment options by prostate cancer (CaP) patients. Among these, the use of dietary supplements and botanical products has been showing a sustained increase in recent years, owing in part to some encouraging pre-clinical and clinical data shown in a limited number of herbal products. Notably, however, the majority of herbal and dietary supplement products have not been rigorously studied with regard to their efficacy. In vitro mechanistic experiments are considered essential preludes and requisites to more lengthy and costly animal and human studies, in that they may provide relevant insights and scientific basis for effects some of these products purportedly might demonstrate. In vitro studies in our laboratory have shown that a polyherbal supplement, Equiguard, exhibits anti-tumor activity against hormone dependent LNCaP cells cultured in both androgen-proficient (FBS) and -deficient (CS-FBS) conditions. Clinically relevant anti-prostate cancer effects of Equiguard are vividly illustrated by growth suppression and down regulated expression of prostate specific genes, respectively, androgen receptor (AR) and prostate specific antigen (PSA). However, the mechanistic bases contributing to these effects have not been well characterized. This communication describes experiments aimed at further understanding growth arrest elicited by Equiguard in LNCaP cells cultured in FBS and CS-FBS conditions. We have focused on aspects of cell cycle control and induction of apoptosis. Regulation of cell cycle progression by Equiguard was analyzed by examining changes in the expression of Rb and cyclins D/E. Using Western blot analysis, we showed that treatment caused inhibition of Rb phosphorylation, which was accompanied by the reduction of cyclins D/E expression, in both culture conditions. Moreover, cells treated with Equiguard and cultured with FBS-supplemented media showed up-regulation of cyclin dependent kinase inhibitor Kip1/p27. These results support the interpretation that suppression of Rb phosphorylation mediated the observed growth arrest induced by Equiguard under androgen-proficient condition. In contrast, Equiguard treated cells cultured in CS-FBS had lowered expression of the Kip1/p27, suggesting that different control mechanisms, possibly evoked by changes in cellular microenvironments, contributed to growth suppression by Equiguard. The growth suppressive effects of Equiguard in both culture conditions were also evaluated with respect to induction of apoptosis. While Equiguard elicited apoptosis was accompanied by an increase in the level of cytosolic cytochrome c, the relative accumulation of cytochrome c in the cytosol was unaffected by culture conditions. These results suggest that the ability to trigger apoptosis as one aspect of the control of cell growth by Equiguard is integrally linked to the release of cytochrome c, by a mechanism largely independent of the presence of androgens. PMID- 15547721 TI - Preventive effect of fermented brown rice and rice bran on N nitrosomethylbenzylamine-induced esophageal tumorigenesis in rats. AB - Modifying effect of fermented brown rice by Aspergillus Oryzae (FBRA) during the initiation or post-initiation phase of N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumorigenesis was investigated in rats. Five-week-old male F344 rats were divided into 7 groups, and groups 1-5 were given subcutaneous injections of NMBA (0.5 mg/kg body weight/injection 15 times) for 5 weeks starting at 7 weeks of age. Groups 2 and 4 were fed the diet containing 5 and 10% FBRA during the initiation phase, respectively, whereas groups 3 and 5 were fed these diets during the post-initiation phase. Group 6 was given the diet containing 10% FBRA throughout the experiment, and group 7 was kept on the basal diet alone and served as an untreated control. Incidence and multiplicity of esophageal neoplasms of group 1 (NMBA alone) were 89% and 1.63+/-1.01/rat, respectively. Those of groups 3 (65%, 1.04+/-1.04) and 5 (58%, 0.77+/-0.86) were significantly less than those of group 1. Furthermore, the incidence and multiplicity of esophageal preneoplastic lesions (dysplasia) of group 5 were less than those of group 1. Post-initiation feeding of 10% FBRA significantly decreased BrdU incorporation in the non-lesional esophageal tissues when compared to that of the control. In addition, the analysis of expression levels of phase I enzymes of livers at the termination of experiment showed no clear differences among the groups. These observations indicate for the first time that FBRA inhibits NMBA induced esophageal tumor development in rats possibly through inhibition of cell proliferation in the post-initiation phase, and suggest that FBRA is a promising dietary agent for prevention of human esophageal cancer. PMID- 15547722 TI - The piperidine nitroxide Tempol potentiates the cytotoxic effects of temozolomide in human glioblastoma cells. AB - Temozolomide (TMZ) is a methylating agent with promising antitumor efficacy for the treatment of melanomas and intermediate-grade gliomas. Unfortunately, its use in the management of high-grade gliomas (glioblastomas) is limited by multifaceted resistance mechanisms. The aim of this study was to evaluate the possibility to improve the cytotoxic response of two human glioblastoma cell lines, U87MG and U373MG, to TMZ by the use of Tempol (TPL), a low molecular weight piperidine nitroxide that has been shown to inhibit in vitro and in vivo growth of murine glioma cells. To this purpose, we used two different schedules for the combined exposure to the two agents. Our data indicate that TPL synergizes with TMZ in both U87MG and U373MG cells for both schedules tested. This effect is accompanied by an increase in apoptotic cell death and by changes in the expression of genes involved in control of the apoptotic process. TPL was also observed to induce a cell-type specific decrease in GSH levels and in GSH related enzyme activities that could contribute to its sensitizing effect. PMID- 15547723 TI - The effect of G-CSF on the in vitro cytotoxicity of cytarabine and fludarabine in the FLAG combination in pediatric acute myeloid leukemia. AB - The combination of fludarabine, cytarabine (ara-C) and G-CSF (FLAG) is routinely used in the treatment of acute myeloid leukemia (AML). In this study we characterized the interactions between fludarabine, ara-C and G-CSF in vitro using AML blasts. Exposure to G-CSF alone resulted in a higher leukemic cell survival (LCS), which might be indicative of increased proliferation. The LCS decreased significantly from 69.7 to 54.0% when blasts were exposed to G-CSF 21 h prior to incubation with ara-C (p=0.01). In contrast, LCS increased significantly (from 55.6 to 69.0%; p=0.04) after sequential exposure to G-CSF and fludarabine. Exposure to 4 combinations of fludarabine (4 h; 0.14 microM and 0.55 microM) and ara-C (96 h; 0.21 and 0.82 microM) (FLA) resulted in additive cytotoxicity. The triple combination (FLAG), 21 h 5 microM G-CSF followed by 4 h fludarabine (0.14 and 0.55 microM) and finally ara-C (0.21 and 0.82 microM) for 96 h also resulted in an additive cell kill. In conclusion, these data support the clinical use of G CSF in combination with ara-C, and the combination of ara-C and FLA. Pre-exposure to G-CSF before FLA (FLAG) did not result in increased cytotoxicity in our experiments, indicative of similar anti-leukemic activity. PMID- 15547724 TI - Biological activity of the S-adenosylmethionine decarboxylase inhibitor SAM486A in human breast cancer cells in vitro and in vivo. AB - The antitumor activity of the S-adenosylmethionine decarboxylase (SAMDC) inhibitor SAM486A in human breast cancer cells was investigated. Our in vitro study focused on testing the effects of SAM486A on the proliferation, clonogenicity, invasiveness, cell signaling and PA levels of hormone-independent MDA-MB-435 human breast cancer cells. We also investigated the antitumor action, effects on polyamine pools and tolerability of SAM486A administered to nude mice carrying MDA-MB-435 xenografts. SAM486A suppressed anchorage-independent and dependent growth and invasiveness of breast cancer cells and the inhibition of cell growth was associated with suppression of spermine synthesis. Combined administration of SAM486A and alpha-difluoromethylornithine (DFMO), a selective inhibitor of ornithine decarboxylase (ODC), exerted greater antiproliferative and anti-invasive effects and induced an overall greater suppression of cellular PA levels than the individual treatments. Both SAM486A and DFMO increased phosphorylation of STAT-1, -3, ERK1/2 and p38, thus indicating activation of both STAT signaling and the MAPK pathway. SAM486A (1 mg/kg) significantly suppressed the growth and spermine levels of established MDA-MB435 breast tumors in nude mice. SAM486A exerts a potent antitumor action in MDA-MB-435 breast cancer cells both in vitro and in vivo. Inhibition of cellular spermine is consistently observed with SAM486A treatment and may mediate its antitumor action. Combination treatment with DFMO may allow the use of lower and, hence, less toxic doses of each compound with preservation of optimal therapeutic effect. The role of activation of STAT signaling and the MAPK pathway in the antitumor action of SAM486A remains to be determined. PMID- 15547725 TI - Extracellular signal-regulated kinase induces cyclin D1 and Cdk-2 expression and phosphorylation of retinoblastoma in hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common malignancies in Southeast Asia. Hyperphosphorylation of retinoblastoma (pRB) by cyclin/CDKs in G1/S transition is required for its inactivation and cell cycle progression. In the present study, we report that phosphorylation of pRB at Ser780 and Ser795 was detected in 71% (33 of 46) and 63% (29 of 46) of HCCs examined respectively. pRB protein was undetectable in 13% (6 of 46) of HCCs examined. Phosphorylated pRB was localized in the nuclei of hepatocarcinoma cells. Benign hepatocytes exhibited very weakly or no nuclear staining for phosphorylated pRB. Over expression of E2F-1, cyclin D1, Cdk-2, Cdk-4 and cyclin A was found in 64% (30 of 46), 43% (26 of 46), 28% (11 of 46), 71% (33 of 46) and 63% (29 of 46) of HCCs examined respectively and this was correlated with elevation of ERK. Treatment of HepG2 cells with MEK1/2 inhibitor U0126 resulted in cell cycle arrest, downregulation of cyclin D1 and Cdk-2 expression and inhibition of pRB phosphorylation at Ser780 and Ser795. Ectopic expression of activated MEK1 in HepG2 cells increased cyclin D1 and Cdk-2 expression, phosphorylation of pRB at Ser780 and Ser795, and percentage of cells in S phase. Our data indicate that activated ERK plays an important role in cyclin D1 and Cdk-2 expression and phosphorylation of pRB at Ser780 and Ser795 in liver cancer cells. PMID- 15547726 TI - Synergistic induction of apoptosis in neuroblastoma cells using a combination of cytostatic drugs with interferon-gamma and TRAIL. AB - The majority of high-risk neuroblastomas lack the expression of caspase-8 due to gene silencing which suggest a mechanism for the selection of tumour cells that are refractory to multiple cytotoxic drugs including tumour necrosis factor related apoptosis-inducing ligand (TRAIL). Inhibitors of DNA methyltransferases and IFN-gamma induce expression of caspase-8, and sensitise some neuroblastoma cells to TRAIL-mediated apoptosis. Here we demonstrate that a combination of cytostatic drugs with IFN-gamma and TRAIL synergistically induces neuroblastoma cell death, which may have implications for future therapy of children with neuroblastoma. Treatment of neuroblastoma cells with IFN-gamma induced caspase-8 expression in all cell lines investigated. In five of the neuroblastoma cell lines (SHEP-1, SK-N-AS, SK-N-FI, SH-SY-5Y and Kelly), IFN-gamma promoted TRAIL mediated cleavage of caspase-8, initiating a caspase cascade involving caspase-7 and PARP followed by apoptosis. IFN-gamma-mediated facilitation of apoptosis was inhibited by the pan-caspase inhibitor zVAD-fmk and the caspase-8 specific inhibitor zIEDT-fmk, indicating an important role of caspase-8 in mediating sensitation by IFN-gamma in neuroblastoma cells. In three of the cell lines [SK-N BE(2), SK-N-DZ and IMR-32] caspase-8 expression was induced by IFN-gamma, but the cells were still resistant to TRAIL-mediated apoptosis. The pattern of basal TRAIL receptor expression, decoy receptors, FLIP and FADD could not be correlated with resistance or sensitivity to TRAIL-induced apoptosis. Importantly, treatment of neuroblastoma cell lines with cytostatic drugs increased apoptosis in the TRAIL-sensitive cell lines whereas the resistant cell lines were susceptible to TRAIL-mediated apoptosis in the presence of the anticancer drugs. The mechanism of the increased susceptibility to apoptosis might results from drug-mediated up regulation of the death receptors DR4 and DR5. PMID- 15547727 TI - Ionizing radiation induces alterations in cellular proliferation and c-myc, c-jun and c-fos protein expression in breast epithelial cells. AB - The identification of genes involved in breast cancer is of critical importance in understanding the pathogenesis of the disease. Expression of the nuclear proto oncogenes, c-myc, c-jun and c-fos, are indicative of early response events during cellular proliferation. Among them, the c-myc oncogene has been found frequently over-expressed in breast cancer. In vitro systems allow us to test the sensitivity of human breast epithelial cells to different carcinogens, including ionizing radiation. The aim of this work was to define whether these oncogenes play a functional role in radiation-induced transformation of human breast epithelial cells. We examined: a) the spontaneously immortalized MCF-10F cell line, b) clones derived from these cells treated with the carcinogen benzo(a)pyrene (BP) and then transfected with c-Ha-ras-oncogene, followed, c) by a single 3 Gy dose of gamma-rays. Protein expressions were analysed by Western immunoblot assays. Results indicated that 3 Gy dose of gamma-ray decreased the expression of these oncoproteins in the MCF-10F cells (ranging from 23 to 80%). In BP1, non-tumorigenic MCF-10F cells, radiation induced an even sharper decrease in the oncoprotein levels (ranging from 50 to 100%) relative to their non irradiated controls. In contrast, in BP1-E tumorigenic cell line radiation increased the expression in 68-80% of c-myc, c-jun and c-fos protein expression relative to non-irradiated control. Furthermore, radiation increased c-my, c-jun and c-fos protein expression in the c-Ha-ras-3 Gy cell line relative to non irradiated control cell line (ranging from 45 and 120%). Interesting, among the tumorigenic MCF-10F cells previously exposed to both BP and c-Ha-ras (BP1-Tras-3 Gy cell line), radiation increased the c-myc, c-jun, c-fos protein expression by more than 120% relative to the non-irradiated controls. In can be concluded that the MCF-10F model of breast carcinogenesis allows us to examine various aspects of regulations in gene expression and can provide us the basis for understanding the process of breast cancer. PMID- 15547728 TI - Antibody-mediated transduction of p53 selectively kills cancer cells. AB - Some human cancers are caused by functional defects in p53 that are restored by gene therapy with wild-type p53. To circumvent the use of viral vectors, we reconstituted cancer cell lines with p53 by protein transduction. A fusion protein was produced from cDNA constructed from the Fv fragment of an antibody that penetrates living cells and wild-type p53 (Fv-p53). Fv-p53 penetrated and killed cancer cells that do not express p53. Additionally, Fv-p53 killed cancer cells that were malignant as a result of mutations within p53, nuclear exclusion of p53 and over-expression of MDM2. Non-specific toxicity was excluded by showing that Fv-p53 penetrated but did not kill primary cells and cancer cells unresponsive to p53. Fv fragments alone were not cytotoxic, indicating that killing was due to transduction of p53. Fv-p53 was shown to penetrate cancer cells engrafted in vivo. These results support continued efforts to evaluate the potential efficacy of Fv-p53 for the treatment of certain cancers in vivo. PMID- 15547729 TI - KIF27 is one of orthologs for Drosophila Costal-2. AB - Signals of Hedgehog family proteins (SHH, IHH and DHH) are transduced through Patched family receptors (PTCH1 and PTCH2) and Smoothened (SMO) to GLI family transcription factors (GLI1, GLI2 and GLI3). SHH plays a key role in development and progression of pancreatic cancer, gastric cancer, basal cell carcinoma, and brain tumors. Drosophila Costal-2 (Cos2) is implicated in the Hedgehog pathway through the interaction with Smoothened (Smo), Cubitus interruptus (Ci), Fused (Fu), and microtubule; however, mammalian ortholog of Drosophila Cos2 remained to be identified. Here we identified and characterized human ortholog of Drosophila Cos2 by using bioinformatics. Full-length Drosophila Cos2 was most homologous to human KIF27, followed by mouse Kif7, and other KIF family members. KIF27 gene at human chromosome 9q22.1 and KIF7 gene at human chromosome 15q26.1 were paralogs within the human genome. Phylogenetic analysis revealed that KIF27, Kif7, KIF4A, KIF4B and KIF21A constitute the KIF27 subfamily among mammalian Kinesin family. Drosophila Cos2 protein consists of Kinesin motor (KISc) domain, Ci-binding domain, and Smo-binding domain. KIF27 itself shared the common domain structure with Drosophila Cos2, while other members of KIF27 subfamily shared partial domain structure with Drosophila Cos2. These facts indicate that KIF27 is one of mammalian orthologs for Drosophila Cos2. PMID- 15547730 TI - Characterization of KIF7 gene in silico. AB - Sonic hedgehog (SHH), Indian hedgehog (IHH) and Desert hedgehog (DHH) are Hedgehog family ligands, which play key roles in embryogenesis and carcinogenesis, especially in pancreatic cancer and gastric cancer. Drosophila Costal-2 (Cos2) is a Hedgehog signaling regulator, interacting with Smoothened (Smo), Cubitus interruptus (Ci), Fused (Fu) and microtubule. KIF27 is one of the mammalian orthologs for Drosophila Cos2. KIF7 gene is the paralog of KIF27 gene; however, complete coding sequence (CDS) of KIF7 remains elusive. Here we characterized human KIF7 gene by using bioinformatics. Human LOC374654 cDNA (NM_198525.1) was a 5'-truncated partial KIF7 cDNA, corresponding to the nucleotide position 1418-4538 of KIF7 complete CDS. KIF7 gene, consisting of 19 exons, was found to encode a 1343-amino-acid KIF7 protein. Human KIF7 showed 43.6% total-amino-acid identity with human KIF27. Kinesin motor catalytic (KISc) domain (codon 15-347 of KIF7) and novel KIF7-KIF27 homologous (KIF727H) domain (codon 699-1196 of KIF7) were well conserved between KIF7 and KIF27. Three coiled coil regions were located within the KIF727H domain. KIF7 and KIF27 were N-type Kinesins with the KIF727H domain. KIF7 mRNA was expressed in embryonic stem (ES) cells, melanotic melanoma and Jurkat T cells, while KIF27 mRNA was expressed in testis, pancreatic islet, germ cell tumors and Jurkat T cells. This is the first report on the characterization of the KIF7 gene as well as on the identification of KIF727H domain. PMID- 15547731 TI - Plasminogen activator inhibitor type-1 mutants regulate angiogenesis of human umbilical and lung vascular endothelial cells. AB - Plasminogen activator inhibitor type-1 (PAI-1) is considered one of the key regulators of tumor invasion, metastasis, as well as cancer-related angiogenesis. The literature suggests that PAI-1 plays a dual role in these processes because it inhibits plasmin-originated proteolysis and binds to vitronectin or integrins. Stimulation or inhibition of angiogenesis largely depends on which of these elements PAI-1 interacts. Wild PAI-1 converts quickly into its latent, inactive form and loses its anti-proteolytic activity, but still binds to vitronectin and integrins. Thus we constructed PAI-1s with extended half-life to prolong their anti-proteolytic activity. We have analyzed the effects of sprout formation inhibition by PAI-1s on two functionally different endothelial cell (EC) systems, human umbilical vein endothelial cells (HUVEC), expressing moderate amounts of urokinase (uPA), and human lung microvascular endothelial cells (HLMVEC), expressing high amounts of this enzyme. We have used wild-type PAI-1 (wPAI-1) (t(1/2) = 1.6 h) and PAI-1 cysteine mutants (CysPAI-1) characterized by their prolonged half-life time (hDbetaT) (t(1/2) = 63.6 h and t(1/2) = 7,000 h). We have observed a significant inhibitory dose-dependent effect exerted by the CysPAI-1s on sprout formation by HUVEC and HLMVEC cells. The inhibition rate was considerably stronger in lung capillary cell cultures and significantly more pronounced for CysPAI-1 mutants with longer anti-uPA activity (betaT). wPAI-1 with a short anti-proteolytic half-life has induced sprout formation in HUVEC, but not in HLMVEC cultures. This difference in behavior was most likely related to the presence of excessive amounts of uPA in HLMVEC cells and the known mechanism of clearing PAI-1/uPA/uPAR complexes from the cell surface. A less efficient system of HUVEC cells might give wPAI-1 the chance to interact with non proteolytic pathways of angiogenesis stimulation. We conclude that while the anti proteolytic properties of PAI-1 constructs are preserved, these proteins inhibit angiogenesis and inhibitory activity dominates over any stimulatory effects of PAI-1. PMID- 15547732 TI - Relationship between immunohistochemical evaluation of thymidylate synthase and proliferating cell nuclear antigen labeling index in gastrointestinal carcinoma. AB - We undertook this study to clarify the effect of immunohistochemical expression of thymidylate synthase (TS) on proliferative activity of carcinoma lesions in patients with gastrointestinal carcinoma. TS was immunohistochemically evaluated in 53 gastric carcinoma and 51 colorectal carcinoma patients using anti-TS polyclonal antibody. Proliferative activity represented by proliferating cell nuclear antigen (PCNA) labeling index (LI) was also immunohistochemically estimated using monoclonal antibody PC10. Then, the correlation between TS expression and PCNA LI was investigated. Both in gastric and colorectal carcinoma, the PCNA LIs of the high-TS group were significantly higher than those of the low-TS group. In gastric carcinoma, the PCNA LIs of the high-TS group were higher than those of the low-TS group in differentiated adenocarcinoma, in the depth of mucosal and/or submucosal layer, in cases without lymph node metastasis, and notwithstanding lymphatic or venous invasion. In colorectal carcinoma, PCNA LIs of the high-TS group were higher than those of the low-TS group in well differentiated adenocarcinoma, in the depth of serosa, in cases with lymph node metastasis, in cases with lymphatic invasion, and notwithstanding venous invasion. Immunohistochemical expression of TS was correlated with the proliferative activity represented by PCNA LI, but was not identical with PCNA LI. PMID- 15547733 TI - Suppressive effect of CPT-11 on rat esophageal tumorigenesis induced by N nitrosomethylbenzylamine. AB - Irinotecan hydrochloride (CPT-11) is a potent anti-cancer drug with suppressive effects against gastric and colorectal cancers. However, no evaluation of CPT-11 for esophageal squamous cell carcinoma has been performed in vivo. In this study, we examined the tumor suppressive effects of CPT-11 on N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal tumorigenesis. One hundred and fourteen rats were divided into six groups. Rats in groups 1-5 were treated with subcutaneous injection of NMBA at a dose of 0.5 mg/kg 3 times per week for 5 weeks. Rats in group 6 served as a control. Rats in groups 2 and 3 were treated with intragastric (i.g.) administration of CPT-11 at doses of 20 and 40 mg/kg, respectively, once a week simultaneously with NMBA-initiation up to the end of the experiment. Rats in groups 4 and 5 were treated with i.g. administration of CPT-11 at doses of 20 and 40 mg/kg, respectively, once a week after the NMBA initiation period up to the end of the experiment. The incidence of papilloma and hyperplasia in the esophagus showed no difference between NMBA-treated groups. However, the multiplicity of hyperplasia was significantly reduced in all CPT-11 administration groups. Immunohistochemically, expression of proliferating cell nuclear antigen (PCNA) was decreased on carcinogen-exposed squamous epithelium and preneoplastic lesions, although no significant differences were detected in the expression of single-strand DNA (ssDNA) and p53. These data suggest that CPT 11 has suppressive effects for esophageal tumors in the early step of the multistep process of carcinogenesis through antiproliferative mechanism. PMID- 15547734 TI - Amplicon profiling reveals cytoplasmic overexpression of MUC1 protein as an indicator of resistance to platinum-based chemotherapy in patients with ovarian cancer. AB - Chromosomal gains of 1q21-q22 and 13q12-q14 were closely related with the chemoresistance of patients with ovarian cancer in our previous CGH (comparative genomic hybridization) study. We conducted the present study to determine the amplified genes on chromosome 1q. Comparisons of relative copy numbers of clinically platinum-sensitive ovarian tumors (CR-group, n=14) and platinum resistant tumors (PD-group, n=14) were carried out using real-time PCR from ten different gene loci on chromosome 1q. Increased copy numbers were frequently observed in PD-group tumors, especially in the region between WI-8123 and MUC1. Relative copy number of MUC1 over 1.5 was observed in 13 (92%) of 14 PD-group tumors and 3 (21%) of 14 CR-group tumors (p<0.05). Moreover, cytoplasmic expressions of MUC1 protein were significantly higher in PD-group than those in CR-group (p<0.01). We concluded that the cytoplasmic overexpression of MUC1 might be an indicator of resistance to platinum-based chemotherapy and a prognostic marker in ovarian cancer. PMID- 15547735 TI - Induction of apoptosis by maackiain and trifolirhizin (maackiain glycoside) isolated from sanzukon (Sophora Subprostrate Chen et T. Chen) in human promyelotic leukemia HL-60 cells. AB - We have investigated the effects of maackiain and trifolirhizin (maackiain glycoside) isolated from sanzukon (Sophora Subprostrate Chen et T. Chen) on DNA of human promyelotic HL-60 leukemia cells. It was found that extent of induction of apoptosis by maackiain was larger than that by trifolirhizin in human leukemia HL-60 cells. Morphological changes showing apoptotic bodies were observed in the HL-60 cells treated with maackiain and trifolirhizin. The fragmentations of DNA by maackiain and trifolirhizin to oligonucleosomal-sized fragments that is a characteristic of apoptosis was observed to be concentration- and time-dependent in the HL-60 cells. The data of the present study show that the suppressions by maackiain and trifolrhizin of growth of the HL-60 cells result from the induction of apoptosis by these compounds, and that the extent of growth suppression and induction of apoptosis by maackiain was greater than that by the glycoside (trifolirhizin). PMID- 15547736 TI - Expression of survivin mRNA associates with apoptosis, proliferation and histologically aggressive features in hepatocellular carcinoma. AB - The clinical impact of survivin on human cancer pathogenesis and prognosis has been investigated. To clarify the clinical effect of survivin on tumor behavior and prognosis of patients with hepatocellular carcinoma (HCC), the expression of survivin mRNA in 40 samples of HCC tissue and matched-adjacent liver tissue, as well as 7 healthy hepatic tissue samples were measured by a real-time reverse transcription polymerase chain reaction. The expressed level of survivin mRNA (log copies/microg total RNA) in healthy liver tissue was 1.95+/-0.44, in morbid liver tissue adjacent to the tumors was 4.79+/-0.96, and in HCC tissue was 5.87+/ 0.73 (values are mean +/- SD and P<0.001). The amount of survivin mRNA in HCC tissues correlated negatively with the apoptotic indices (r=-0.573 and P<0.001) and correlated positively with the proliferation indices (r=0.433 and P=0.005). Expression of survivin was significantly related with histologic grade (P=0.011) and pathological tumor stage (P=0.017). Patients with HCC tumors that had a large amount of survivin mRNA (> or = mean) had lower survival rate (P=0.030), but multivariate analysis showed only Ki-67 labeling index, histologic grade, and pathologic T stage to be the independent prognosticators. These findings indicate that survivin is associated with reduced tumor cell apoptosis, increased tumor cell proliferation, and histologically aggressive tumor features, and may play an important role in tumor progression of HCC. However, further examination is needed to clarify its predictive significance for HCC patients. PMID- 15547737 TI - Intrinsic radiosensitivity by metallothionein expression has no great influence on clinical radiosensitivity in esophageal carcinoma. AB - Several studies have shown that macroscopically infiltrative type of esophageal carcinoma generally has a poorer local response to radiation therapy than that of localized type. The aim of this study was to determine the role of metallothionein (MT) as a radioprotective agent in the difference of clinical radiosensitivities in esophageal carcinoma. A total of 45 surgically resected esophageal carcinoma tissues [20 macroscopically localized type without preoperative treatment (PT), 20 macroscopically infiltrative type without PT and 5 macroscopically infiltrative type with PT] were stained for MT by immunohistochemistry and were analyzed. MT expression level of macroscopically localized type of esophageal carcinoma was significantly higher than that of infiltrative type without PT (P=0.026) and was significantly higher than that of infiltrative type with PT (P=0.024). No significant difference was found between MT expression levels in infiltrative type of esophageal carcinoma without PT and that with PT. The results of this study suggest that there is less possibility that clinical radioresistance of the tumor is due to MT expression in esophageal carcinoma and also suggest that there is less possibility that MT synthesis is induced by fractionated irradiation of a moderate dose or by an anti-cancer agent during a course of treatment. PMID- 15547738 TI - P-glycoprotein 170 expression and function as an adverse independent prognostic factor in childhood acute lymphoblastic leukemia. AB - Little is known about the prognostic role of multidrug resistance (MDR) in newly diagnosed childhood acute lymphoblastic leukemia (ALL). P-glycoprotein 170 (MDR1), a cellular drug efflux pump, is thought to be one of the major causes of MDR. The aim of this retrospective study was to evaluate in 85 children with ALL the impact of the MDR1 product of the mdr-1 gene on the achievement of complete remission (CR) and outcome. MDR1 protein expression was performed by immunocytochemistry (ICC), and flow cytometry (FC). MDR1 functional activity was performed by a rhodamine (Rhd)-123 efflux test with or without verapamil. All patients enrolled in our study were treated with AIEOP ALL 91-95 protocols. At diagnosis, 40 patients (47%) expressed MDR1 protein at significant levels, and 45 (53%) were MDR1 negative. Forty-three of the latter patients were also negative for MDR1 function, while 34/40 (85%) patients MDR1 positive preserved the function. Rhd-123 efflux was inhibited by the MDR modulator verapamil in 12/40 (30%) patients. After induction treatment, CR was achieved in 77/85 children (90.6%). All patients who did not achieve CR were MDR1 positive. Twenty-nine patients relapsed, 17 (58.6%) of whom were MDR1 positive. The 10-year overall survival (OS) rate, and disease-free survival (DFS) for MDR1 negative patients compared to MDR1 positive patients were 75.7% versus 54.8%, and 67.5% versus 46%, respectively. The 10-year event-free survival (EFS) rate was significantly higher (67.5% versus 36.8%) in the MDR1 negative group compared with the MDR1 positive population (p=0.001). Multivariate analysis showed that only EFS was independent of age, WBC count, immunophenotype, FAB subtype and prednisone response (p=0.019). Our results, derived from a monocentric study, demonstrate that MDR1 expression in childhood ALL is an independent adverse prognostic factor on outcome, and could be a useful biological marker of response in these patients. Moreover, MDR1 function was also a predictor of response, but only in univariate analysis. PMID- 15547739 TI - Nitric oxide production in fluoro-edenite treated mouse monocyte-macrophage cultures. AB - In the present study, we investigated the involvement of NO in the cytotoxic and genotoxic effects caused by fluoro-edenite in mouse monocyte-macrophage cell line J774. Fluoro-edenite is a new asbestos-like amphibole present in the benmoreitic lavas recently extracted from stone quarries in Biancavilla, a village located in the Etnean Volcanic Complex (Catania, Italy) of eastern Sicily, in which an epidemiological survey evidenced a cluster of cases of the mortality due to pleural mesothelioma. Fluoro-edenite appears as a probable carcinogenic agent. Nitrite and nitrate concentration (NO) in the supernatant was quantified by colorimetric assay based on the Griess reaction and iNOS (inducible nitric oxide synthetase) expression was determined by immunostaining in mouse monocyte macrophage cell line J774 treated with different concentrations of fluoro-edenite (5, 50 and 100 microg/ml) for 24, 48, 72 and 96 h. Parallel experiments were performed treating the cultures also with lipopolysaccharides (LPS), used as inflammation-inducing molecule. In our experimental conditions, fluoro-edenite did not modify the level of NO and the expression of iNOS at the experimental used concentrations for 24, 48 and 72 h. These parameters were significantly modified at the higher doses (50 and 100 microg/ml) of fluoro-edenite for 96 h and were further more increased, in concentration- and time-dependent manner, when cell cultures were treated with fluoro-edenite and LPS. These findings provide convincing evidence that NO is involved in the fluoro-edenite-induced cytotoxicity and geno-toxicity in mouse monocyte-macrophage cell line J774 when the fiber remain for longer times and particularly in cultures treated with LPS, demonstrating that inflammatory disorders appear to increase the risk for lung cancer induced by fluoro-edenite probably by the involvement of NO. PMID- 15547740 TI - The role of hMSH3 and hMSH6 in ovarian endometrioid carcinoma and relationship with microsatellite instability phenotype. AB - Frequent frameshift mutations in the DNA mismatch repair genes hMSH3, and hMSH6, have been reported in colorectal and endometrial cancers with microsatellite instability, however, it is unclear whether they are similarly altered in ovarian endometrioid carcinoma. In this study, we examined frequency of frameshift mutation and protein expression in hMSH3 and hMSH6 in ovarian endometrioid carcinoma with or without microsatellite instability. Only 1 frameshift mutation of the 16 cases with microsatellite instability-high phenotype (6%) was detected in poly(A)8 tract of the hMSH3, but not in poly(C)8 tract of hMSH6 genes. In addition, none of the 6 microsatellite instability-low or 20 microsatellite stable tumors showed mutations in these regions in either gene. These results indicate that mutations in the mono-nucleotide tracts of hMSH3 and hMSH6 are infrequent in ovarian endometrioid adenocarcinomas, other mechanisms may play a more important role in the development of these tumors. PMID- 15547741 TI - Assessment of K-ras, Smad4 and p53 gene alterations in colorectal metastases and their role in the metastatic process. AB - To date there is no genetic marker that gives accurate information on the prognostic impact for patients with colorectal cancer. A particular clone, not detected in the tumor, could be responsible for the metastatic process. To overcome this problem, genetic alterations were analyzed in metastatic tissues from 58 patients who developed metastases after curative surgery for colorectal cancer. K-ras, p53 and Smad4 alterations were observed in respectively 38, 60 and 27% of the metastases. These frequencies are similar to the ones reported in primary colorectal tumors. Thus, these genetic alterations cannot be used as prognostic biomarkers in patients with colorectal cancer. The metastases were stratified into 3 groups, according to the metastatic localization. K-ras mutations were detected in respectively 75, 26 and 11% of the distant, peritoneal and liver metastases. Loss of Smad4 expression was observed more frequently in the liver (62%) than in other metastases (13%). These results suggest that the genetic changes of the tumor cells indicate the location of the metastases and thus, the route of metastatic spread. PMID- 15547742 TI - Demonstration of human telomerase reverse transcriptase by in situ hybridization in lung carcinoma. AB - Telomerase activity is present in most malignant tumors and provides a mechanism for unlimited replication of neoplastic cells. This study was undertaken to investigate the expression of the gene encoding human telomerase reverse transcriptase (hTERT), the telomerase catalytic subunit gene in lung carcinoma, by in situ hybridization (ISH). hTERT is associated with telomerase activity, and overexpressed in most lung carcinomas. We assayed hTERT gene expression by ISH to study telomerase activity in lung carcinomas of 27 patients, and compared these results with those of telomerase activity by telomeric repeat amplification protocol (TRAP) assay. TERT gene expression by ISH was detected mainly in the nuclei of lung carcinoma cells. Some specimens showed a significant expression in only infiltrating lymphocytes. hTERT gene expression by ISH was found in 16 of 27 (59%) cases. On the other hand, telomerase activity by TRAP assay was detected in 21 of 27 (78%) cases. In 7 cases, TRAP assay detected telomerase activity, but ISH did not detect hTERT expression. TRAP assay might detect telomerase activity in not only carcinoma cells, but also in infiltrating lymphocytes. Our findings therefore suggest that ISH-based analysis of hTERT gene expression is superior to TRAP assay as a means of determining telomerase status during carcinogenesis. PMID- 15547743 TI - Methyl jasmonate induces apoptosis through induction of Bax/Bcl-XS and activation of caspase-3 via ROS production in A549 cells. AB - Jasmonates are plant lipid derivatives, similar to mammalian eicosanoid, that play a critical role(s) in plant defenses against herbivores and pathogens through up-regulating the expression of defense-related genes. Recently, jasmonates were shown to induce cell cycle arrest or apoptosis in human leukemia, prostate and breast cancer cells, but not in normal lymphocytes, suggesting that the chemicals can be used as a novel class of anti-cancer drugs. In the present study, we examined the molecular mechanism that contributes to methyl jasmonate induced apoptosis. Herein we show that methyl jasmonate induces apoptosis through induction of Bax/Bcl-XS and activation of caspase-3 via reactive oxygen species production in A549 human lung adenocarcinoma cells. PMID- 15547744 TI - Cdx2 expression in pancreatic tumors: Relationship with prognosis of invasive ductal carcinomas. AB - We have previously reported that Caudal-related homeobox gene 2 (Cdx2) is a useful prognostic intestinal phenotypic marker for advanced gastric cancers. In this study, we examined Cdx2 expression and phenotype in pancreatic tumors. We evaluated 19 mucinous cystic tumors (MCTs), 17 intraductal papillary-mucinous tumors (IPMTs), and 41 invasive ductal carcinomas (IDCs) with regard to their gastrointestinal phenotype. The expression of Cdx2 was also assessed immunohistochemically. The lesions were phenotypically divided into 39 gastric (G type), 29 gastric and intestinal mixed (GI type), 3 intestinal (I type), and 6 null (N type) types, independent of the histopathological type. Most of the pancreatic tumors were thus judged to be the positive for gastric phenotypic members. Cdx2 nuclear staining demonstrated a close relation to the intestinal phenotypic expression in all three types (MCTs, IPMTs, and IDCs; p<0.05). In IDCs, Kaplan-Meier analysis of Cdx2 expression showed the Cdx2-positive group to have a significantly better outcome than their negative counterparts (p=0.015). In conclusion, our data suggest that Cdx2 might be necessary for intestinal phenotypic expression even in pancreatic tumor cells. In addition, Cdx2 expression in IDCs may be a novel prognostic marker for patient survival. PMID- 15547745 TI - mRNA and protein expression of PUMA in sporadic colorectal cancer. AB - PUMA is a BH3-only member of the Bcl-2 family, up-regulated by p53 as a response to DNA damage. We have investigated the mRNA expression of PUMA with real-time PCR in 94 colorectal adenocarcinomas and the corresponding normal mucosa. Among them PUMA protein expression was investigated with immunohistochemistry in 23 tumours and 17 corresponding normal mucosa samples. The mRNA expression of PUMA decreased in 4% and increased in 4% of the tumours compared with the normal mucosa. The protein expression of PUMA decreased in 6% and increased in 29% of the tumours compared with the normal mucosa. Decreased PUMA expression in the tumour compared with the corresponding mucosa was correlated with the distal colon and rectum (P=0.02). We did not find any other relationship to clinical or pathological features. We suggest that the changes in PUMA expression may be of minor importance in the development of colorectal cancer. PMID- 15547746 TI - Characterization of stage-specific embryonic antigen-1 expression during early stages of human embryogenesis. AB - Extensive expression of stage-specific embryonic antigen-1 (SSEA-1) has been documented in some animal species, but not in human embryos. In this study, SSEA 1 was detected during human embryogenesis by whole-mount immunohistochemistry. Alkaline phosphatase (Ap) activity was detected to identify human primordial germ cells. SSEA-1 was expressed steadily and restrictedly in some cells/tissues, especially in the nephric duct and nephric tubule (including the pronephric duct and tubule, mesonephric duct and tubule, metanephric tissues) besides embryonic ectodermal cells and yolk sac from 3 to 7 weeks. High level of Ap activity was observed in vessels, part of the mesonephric duct, especially in embryonic primordial germ cells localized in the yolk sac, primitive gut, dorsal mesenteries and genital ridges. No colocalization of AP and SSEA-1 cells was observed. SSEA-1 was expressed in human embryos in a different pattern at early stages compared to that in mouse embryos. It was expressed in the nephric duct, nephric tubule, yolk sac and on the surface of embryonic ectodermal cells of the epidermis, but not in human primordial germ cells. PMID- 15547747 TI - Utility of initial MRI for predicting extent of residual disease after neoadjuvant chemotherapy: analysis of 70 breast cancer patients. AB - This study was performed to evaluate the utility of initial MRI in predicting the extent of residual disease following neoadjuvant chemotherapy (NAC). The study population consisted of 70 patients with breast cancer (unilateral, n=69; bilateral, n=1) (mean age 51 years) who underwent magnetic resonance imaging (MRI) with gadolinium enhancement both before and after NAC. Basic NAC was comprised of cyclophosphamide, pirarubicin, and 5-fluorouracil. MRI features were compared with pathological diagnosis following surgery. MRI features of breast cancer before NAC were classified as either solitary nodular (SN) (n=33) (47%), or multiple nodular and/or unlocalized dendritic (MN/UD) (n=38) (53%). MRI typing was independent of NAC in 68 tumors (SN, n=32; MN/UD, n=36) (96%, p<0.0001). All except one of the 33 SN tumors (97%) displayed negative margins. In addition, 5 of the 33 SN tumors (15%) displayed pathological complete response (pCR). Conversely, all 5 requiring total glandectomy due to wide infiltration and all except one of the 17 (94%) displaying positive margins necessitating extended resection were classified as MN/UD. Only SN-type tumors on initial MRI have the possibility of pCR after NAC. MN/UD tumors could possess margins necessitating expanded excision or total glandectomy. Morphological concepts based on MRI can prove useful in surgical planning and predicting the extent of residual disease after NAC. PMID- 15547748 TI - TU12B1-TY, a novel gene in the region at 12q22-q23.1 frequently deleted in pancreatic cancer, shows reduced expression in pancreatic cancer cells. AB - Loss of heterozygosity (LOH) on 12q is frequently observed in primary pancreatic cancer, as well as in cancers of other tissues such as stomach and germ cells. LOH correlates with poor prognosis in patients suffering from pancreatic cancer. In quest of tumor suppressor genes in this region, we used bacterial artificial chromosome (BAC) clones to construct a contig to cover one of the two targeted regions previously detected in pancreatic cancer; this region, 12B, is no larger than 650-kb between D12S360 and D12S78 at 12q22-q23.1. While constructing a detailed physical map and placing expressed sequence-tags, we identified a novel human gene, TU12B1-TY. This gene consisted of at least 14 exons and harbored an open reading frame possibly encoding a 473 amino-acid protein. A motif prediction program revealed a transmembrane domain in its carboxyl terminus. Expression in human tissues was found in the brain, placenta, skeletal muscle, pancreas, testis, uterus, and small intestine. In 21 pancreatic cancer cell lines analyzed, we found no structural alteration but all of them showed reduced expression. The present results indicate that reduced function of TU12B1-TY may contribute to the development and/or progression of human pancreatic cancer. PMID- 15547749 TI - Hotspot mutations of BRAF gene are not associated with pediatric solid neoplasms. AB - BRAF (v-raf murine sarcoma viral oncogene homolog B1) activating mutations in a high proportion of melanomas and in a small fraction of other cancers have been recently reported. All the presented mutations of BRAF are located in exons 11 and 15, and the hotspot mutation at codon 599 accounts for 87% of BRAF mutations. Because the mutational status is unclear in pediatric solid neoplasms, we screened BRAF mutations comprehensively in our tumor series presented in childhood. Two pairs of primers were designed to amplify exons 11 and exon 15, respectively, and 181 tumor samples (65 neuroblastomas, 23 Wilms tumors, 19 hepatoblastomas, 16 teratomas, 17 rhabdomyosarcomas, 13 ganglioneuromas, etc.) were investigated by PCR-SSCP method. On agarose gel electrophoresis, amplified PCR fragments showed no size-altered changes in exons 11 and 15, and SSCP analysis revealed uniform band patterns in both exons. Subsequent direct sequencing of selected 10 samples confirmed only normal sequences without any nucleotide substitutions. The current study represents the first genetic analysis of the BRAF gene in pediatric solid tumors. Our data suggest that mutations of BRAF gene as a mechanism of tumorigenesis is unlikely to be associated with most childhood neoplasms. PMID- 15547750 TI - Loss of imprinting of PEG1/MEST in lung cancer cell lines. AB - Paternally expressed imprinted gene 1/mesoderm-specific transcript (PEG1/MEST) is an imprinted gene expressed from the paternal allele. Recently, frequent loss of imprinting (LOI) of PEG1/MEST has been reported in lung adenocarcinomas. It is suggested that the LOI may be involved in pathogenesis of lung adenocarcinoma. In the present study, incidence of LOI of PEG1/MEST was examined in lung cancer cell lines, including small cell lung cancer (SCLC). Among 50 cell lines tested, 20 cell lines were heterozygous for the AflIII site of the PEG1/MEST gene. In these heterozygotes, biallelic expression was observed in 9 cell lines (45%), monoallelic in 11 (55%). In cell lines of non-small cell lung cancer (NSCLC), 62% (8 of 13) exhibited biallelic expression. In SCLC, only 1 of 7 cell lines (14%) showed biallelic expression. LOI of PEG1/MEST in the NSCLC cell line is significantly frequent compared with that in SCLC cell lines (p=0.043). This result supports the possibility that LOI may be related to tumorigenesis and malignant transformation, especially in NSCLC. PMID- 15547751 TI - Effects of thymidine phosphorylase levels in cancer, background mucosa, and regional lymph nodes on survival of advanced gastric cancer patients receiving postoperative fluoropyrimidine therapy. AB - Due to the contradictory roles that thymidine phosphorylase (TP) plays in both tumor aggressiveness and fluoropyrimidine activation, its impact on drug responses has yet to be clearly established. Therefore, the present study was performed to clarify the effects of TP levels on the prognosis of gastric cancer patients treated postoperatively with different fluoropyrimidines. A total of 52 gastric cancer patients who underwent gastrectomy from January 1997 to March 1998 were enrolled in the present study. The TP levels in the specimens were assayed by enzyme-linked immunosorbent assay (ELISA). Survival was significantly poorer for the 27 patients with high normal tissue TP activity than for the 25 with low normal tissue TP activity. Normal tissue TP level showed different effects on survival according to the chemotherapy regimen used. While the survival rate was significantly poorer in patients with high normal TP level than in those with low normal TP in the 5-FU group, the rate was almost the same in the 5'-DFUR group. Cox's proportional hazard model revealed that tumor TP was an independent prognostic factor in gastric cancer patients. Since activating and catabolizing enzymes for fluoropyrimidines differ from each other, alterations in gene expression of these enzymes should be useful predictive factors. PMID- 15547752 TI - Gene expression in response to anti-tumour intervention by polysaccharide-K (PSK) in colorectal carcinoma cells. AB - Distant metastasis is one of the major problems in treatment for advanced colorectal cancer. Polysaccharide-K (PSK), or Krestin, a mushroom ingredient, has been used as a chemoimmunotherapeutic agent for the treatment of cancers in Asia for over 30 years. Some studies have reported that PSK prevent distant metastases and improve survival rates by 10-20% in colorectal cancer. However, the mechanism of the interrelated immunomodulatory and direct anti-cancer cell activities of PSK has yet to be elucidated. To investigate the direct effect, we used cDNA microarrays to analyse expression profiles in a human colorectal adenocarcinoma cell line, HCT116, containing the wild-type p53 gene. Expression of 453 genes was significantly altered (142 up-regulated and 311 down-regulated) after 96 h exposure to 500 microg/ml PSK. Under more stringent conditions, 9 genes were up regulated and 36 down-regulated. We then examined the expression of candidate genes in two cell lines, HCT116, and SW480, a cell line with a mutant p53 gene. Our results suggest that PSK may augment anti-tumour action via genes including multidrug resistance protein 3 (MRP3), lymphotactin (Lptn), transgelin (TAGLN), and Pirin, without disturbing cell-cycle progression, and may deserve a large clinical trial in cancer therapy. PMID- 15547753 TI - Long-term survival of patients with advanced colorectal cancer may not be due to the response to chemotherapy. AB - Our primary objective was to determine the median and overall survival and secondarily the response rate to first- and second-line chemotherapy of patients with advanced colorectal metastatic disease. Three-hundred and seventy-nine patients (median age 60 years, range 30-87 years) were enrolled from April 1993 to March 2000. Median follow-up was 6 years (range 3-10 years), until July 2003. All patients were evaluable for survival and 342 were evaluable for response and toxicity. Thirty-seven patients did not undergo chemotherapy. All patients had confirmed histology as well as metastatic disease based on radiological tests. First-line treatment was administered to 342 patients: leucovorin (LV) 30 mg/m2 and 5-fluorouracil (5-FU) 425 mg/m2. Three different combinations were given as second-line treatment during different chronological periods: i) 5-FU, mitomycin C and doxorubicin (FAM); ii) 5-FU and cisplatin (CDDP) and iii) 5-FU, LV and irinotecan (CPT-II). Responses were observed as follows: first-line treatment 16.37%, after FAM 25%, following 5-FU-CDDP 26.83% and after 5-FU-LV-CPT-II, 30.61%. Survival of all patients was as follows: median 25 months (range 16.1 33.9 months). The longest survival was of patients on 5-FU-LV-CPT-II. Median survival of patients with stable disease was 19 months and of untreated patients 12 months. Patients with advanced colorectal cancer have a long median (25 months) and overall survival, despite low responsiveness to chemotherapy. PMID- 15547754 TI - Predicting recurrence and metastasis of stage III/Dukes' C colorectal cancer with lymph node metastasis. AB - This study was designed to compare the prediction of recurrence based on detection of occult neoplastic cells (ONCs) in lymph nodes or using specific criteria to identify patients at high risk of recurrence/metastasis among 105 patients with Dukes' C colorectal cancer. Prediction of recurrence based on the detection of ONCs had a sensitivity of 50.0% (22/44), specificity of 80.3% (49/61), and an accuracy of 65.2%. Prediction of recurrence based on positivity for at least 2 of the 3 high-risk criteria had a sensitivity of 54.5% (24/44), specificity of 83.6% (51/61), and an accuracy of 69.1%. Among the 34 patients with ONCs, prediction of recurrence based on positivity for all 3 high-risk criteria had a sensitivity of 27.3% (6/22), specificity of 91.7% (11/12), an accuracy of 59.5%, and a positive predictive value (PPV) of 85.7% (6/7). These results suggest that the predictive value of ONCs and the high-risk criteria was similar, and that recurrence is likely to occur in patients who fulfill < or =2 of the high-risk criteria. Accordingly, combined use of these parameters may be more effective for the early prediction of recurrence/metastasis to assist in the choice of postoperative systemic chemotherapy. PMID- 15547755 TI - Predicting recurrence and metastasis of Dukes' A primary colorectal cancer with or without proper muscle invasion. AB - This study compared the prediction of recurrence based on detection of occult neoplastic cells (ONCs) in lymph nodes or using 3 criteria to identify high-risk patients among 72 patients who had Dukes' A colorectal cancer with or without proper muscle invasion. Predicting recurrence based on the detection of ONCs had a sensitivity of 40.0% (2/5) and a false-negative rate of 60.0% (3/5), while there was a specificity of 97.0% (65/67) and false-positive rate of 3.0% (2/67), resulting in an accuracy of 68.5%, PPV of 50.0% (2/4), and NPV of 95.6% (65/68). Predicting recurrence based on the presence of at least 2 of the 3 high-risk criteria showed a sensitivity of 60.0% (3/5) and a false-negative rate of 40.0% (2/5), while it had a specificity of 74.6% (50/67) and a false-positive rate of 25.4% (17/67), resulting in an accuracy of 67.3%, PPV of 15.0% (3/20), and NPV of 96.2% (50/52). These results suggest that a prediction based on ONCs was similar to use of the high-risk criteria, with both methods having a high specificity for recurrence/metastasis of Dukes' A colorectal cancer. PMID- 15547756 TI - ErbB2 oncogene antibodies differentiate between the normal and diseased pancreas, and between chronic pancreatitis and pancreatic cancer. AB - Histological differentiation between chronic pancreatitis and pancreatic cancer, especially in biopsy material, remains challenging and the frequent association of 'secondary' chronic pancreatitis (due to ductal obstruction) with pancreatic cancer causes additional diagnostic problems. Our study, using anti-ErbB2 antibodies from Santa Cruz and Dako in tissues from the normal pancreas, chronic pancreatitis and pancreatic cancer showed that these antibodies discriminate between primary chronic pancreatitis and 'secondary' chronic pancreatitis due to pancreatic cancer. Tissues from 28 pancreatic cancer patients, 15 chronic pancreatitis patients and 12 organ donors or early autopsy cases were subjected to immunohistochemical studies using polyclonal ErbB2 antibodies from Santa Cruz and Dako. The Santa Cruz antibody immunoreacted with islet cells in all tissues from the normal pancreas and pancreatic cancer but not in any chronic pancreatitis specimen. The Dako antibody showed a membrane staining of ductal and ductular cells only in chronic pancreatitis cases but in none of the normal or cancer specimens. Moreover, in chronic pancreatitis cases, ductular cells were stained with the Santa Cruz antibody only in the severe form, but not in the mild or moderate form of the disease. The utilized ErbB2 antibodies discriminate between the normal pancreas, chronic pancreatitis and pancreatic cancer. Hence, these antibodies seem to present an additional useful aid in the surgical pathology of pancreatic diseases. PMID- 15547757 TI - Immunohistochemical staining of metallothionein in canine mammary tumors: better survival with higher expression. AB - Metallothioneins (MT) are small proteins able to strongly bind and therefore neutralize heavy-metal, free-radical and other genotoxic compounds. A close relationship between MT and zinc-finger motif-dependent transcription factors has also been shown, with implications on cell proliferation and survival. Human malignant neoplasms (e.g. breast cancer) usually present worse prognosis with increased MT content. We therefore evaluated the immunohistochemical expression of MT in a group of 15 canine mammary gland tumors. There was no relationship between MT immunostaining and histological type or malignancy grading of the lesions. Comparison of MT immunolabelling with the overall survival of the animals revealed that MT overexpression was related to better prognosis, a contrasting finding with the human counter-part. We considered that differential MT gene expression could be responsible for this variation, as observed for some human neoplasms of distinct embryonic origin, but further investigation is required to elucidate this topic. PMID- 15547758 TI - Inactivation of HTB63 human melanoma cells by irradiation with protons and gamma rays. AB - The effects of single irradiation with gamma rays and protons on HTB63 human melanoma cell growth were compared. The exponentially growing cells were irradiated with gamma rays or protons using doses ranging from 2-20 Gy. At 48 h of post-irradiation incubation under standard conditions, cell survival and induction of apoptotic cell death were examined. The best effect of the single irradiation with gamma rays was the reduction of cell growth by up to 26% (p=0.048, irradiation vs. control), obtained using the dose of 16 Gy. The same doses of proton irradiation, having energy at the target of 22.6 MeV, significantly inhibited melanoma cell growth. Doses of 12 and 16 Gy of protons provoked growth inhibition of 48.9% (p=0.003, irradiation vs. control) and 51.2% (p=0.012, irradiation vs. control) respectively. Irradiation with 12 and 16 Gy protons, compared to the effects of the same doses of gamma rays, significantly reduced melanoma cell growth (p=0.015 and p=0.028, protons vs. gamma rays, respectively). Estimated RBEs for growth inhibition of HTB63 cells ranged from 1.02 to 1.45. The electrophoretical analyses of DNA samples and flow cytometric evaluation have shown a low percentage of apoptotic cells after both types of irradiation. The better inhibitory effect achieved by protons in contrast to gamma rays, can be explained considering specific physical properties of protons, especially taking into account the highly localized energy deposition (high LET). PMID- 15547759 TI - Response of T and B lymphocytes in the spleen, lymph nodes and mammary tumors in rats treated with human soluble tumor-associated antigens and commercial human albumin. AB - We showed previously that soluble tumor-associated antigens (sTAA) isolated from breast cancer patients could suppress chemically-induced tumorigenesis in rats in comparison to the effect of commercial human albumin (CHA). Herein we analyze the possible mechanism of those findings. The following groups of mammary tumor bearing rats were used in the studies: i) control rats treated with saline; ii) rats treated with CHA; and iii) rats treated with human sTAA. Different zones of the spleen, regional lymph nodes and tumors and their cellular content (B and T cells) were analyzed using the methods of morphometry and immunohistochemistry. Treatment of tumor-bearing rats with CHA resulted in a significant decrease in the size of the germinal center of the follicles. The number of B lymphocytes in the mantle layer of the follicles, the marginal zone and red pulp decreased significantly. The number of CD8+ T cells also decreased in the marginal zone and red pulp, whereas the number of CD4+ T cells increased in the periarterial lymph sheath (PALS) and the red pulp. Reaction of the spleen to vaccination with sTAA manifested in a significant increase in the size of most areas of the white pulp and in the number of B lymphocytes. In lymph nodes from control rats or those treated with CHA, CD8+ lymphocytes mainly accumulated in the paracortical zone. In rats treated with sTAA, CD8+ lymphocytes accumulated also in the medulla. The number of CD4+ T cells in these rats sharply increased and accumulated mainly in the medulla around the vessels. The total number of lymphocytes was changed differently in different areas of tumors (peripheral vs. at depth). The number of CD8+ cells significantly increased at depth of tumors, and also the ratio in the number of these cells at depth of tumors compared to a periphery increased. No difference was found in response of lymph cells to different types of treatment. All findings indicated a strict antitumor effect of vaccination with the sTAA, which prevents the development of insufficiency of the immune system when an intensive immune reaction takes place. PMID- 15547760 TI - Frequent CYP1A1 expression in gastric cancers and their related lesions. AB - CYP1A1 expression in various tissues of 43 gastric cancer cases was analyzed with frozen tissue array-based immunohistochemical staining (IHC), RT-PCR, Western blot analysis and EROD (7-ethoxyresourfin-O-deethylase) assay. CYP1A1 was detected immunohistochemically in 86% (37/43) of gastric cancers, in both 57% (4/7) of atrophic gastritis and intestinal metaplasia and in 7.7% (1/13) of non cancerous mucosa. RT-PCR and Western blot analysis revealed coincident results with that of IHC and ruled out possible concurrent CYP1A2 expression. EROD assay showed increased CYP1A1/1B1 activity in either cancer or premalignant tissues but not in non-cancerous ones. The frequencies of CYP1A1 expression are significantly different between non-cancerous and premalignant (P<0.025) or cancer groups (P<0.005), suggesting that CYP1A1 is expressed at relatively early stage of gastrocarcinogenesis and exerts its effects throughout the stepwise oncogenic processes. PMID- 15547761 TI - PTEN and DMBT1 homozygous deletion and expression in medulloblastomas and supratentorial primitive neuroectodermal tumors. AB - Medulloblastoma, which accounts for 20-25% of all childhood brain tumors, is defined as a primitive neuroectodermal tumor (PNET) located in the cerebellum. Supratentorial PNET are less frequent than medulloblastoma. But their clinical outcome is worse than in medulloblastomas. Chromosome 10q contains at least 2 tumor suppressor genes that might play a role in brain tumor development: PTEN and DMBT1. The aim of this study was to compare the status of homozygous deletion and expression of PTEN and DMBT1 genes in PNET primary tumor samples and cell lines. Homozygous deletions of PTEN and DMBT1 were studied in 32 paraffin embedded PNET samples (23 medulloblastomas and 9 supratentorial PNET) and in 7 PNET cell lines, by differential PCR and by FISH. PTEN homozygous losses were demonstrated in 7 medulloblastomas (32%) and in no supratentorial PNET, while homozygous deletions of DMBT1 appeared in 1 supratentorial PNET (20%) and in 7 medulloblastomas (33%). No homozygous deletion of PTEN or DMBT1 was detected in any of the PNET cell lines either by differential PCR or by FISH. Expression study of the 2 genes was performed in the 7 PNET cell lines by RT-PCR. One PNET cell line lacked PTEN and DMBT1 expression, while 2 medulloblastoma cell lines did not express DMBT1. Our results add some positive data to the hypothesis that supratentorial PNETs and medulloblastomas might be genetically different. PMID- 15547762 TI - The role of Raf-1 in radiation resistance of human hepatocellular carcinoma Hep G2 cells. AB - Development of radiation resistance is one of the major reasons that cancer cells do not respond to radiotherapy and the mechanism for resistance is still not clear. Two sublines of human hepatocellular carcinoma Hep G2 cells were established from cells that survived two different irradiation regimes, 2 Gy for 10 days or 10 Gy for 2 days, respectively. Using MTT assay, the radiation conditioned cells were found to be more resistant to gamma-irradiation and have a greater extent of potentially lethal damage repair (PLDR) for radiation than the parent cells. By Western blot analysis, the radiation-conditioned cells were found to overexpress Raf-1 which is known to regulate the radiation resistance of cells. Inhibition of Raf-1 expression by antisense oligonucleotides increased the radiation sensitivity of the radiation-conditioned cells while inhibitors of Ras (L744,832), PI3K (LY294002) and p38 (SB203580) had no effect. Moreover, antisense Raf-1 oligonucleotides also decreased the radiation induced PLDR capacity of the radiation conditioned cells. It is therefore suggested that Raf-1 may induce radiation resistance through an increase in radiation induced PLDR capacity in Hep G2 cells. PMID- 15547763 TI - Expression levels of melanoma inhibitory activity correlate with time to progression in patients with high-grade glioma. AB - Melanoma inhibitory activity (MIA) is related to disease progression in patients with malignant melanoma and to invasion and metastasis of melanoma in vivo and in vitro. An alternative splice product termed MIA(splice) was described recently. In addition to melanoma, both proteins are expressed in a substantial subset of high-grade gliomas. We hypothesize that expression levels of both proteins correlate with early tumor progression and parameters of disseminated disease in patients with high-grade glioma. We examined the correlation of expression levels of MIA and MIA(splice) with time to progression and morphological and clinical markers of disseminated disease (defined as multifocal occurrence, gliomatosis, invasion or metastasis) in a series of 24 newly-diagnosed human high-grade gliomas. Homogenates of surgical specimens, cell cultures and blood samples were analyzed. Significant levels of MIA and MIA(splice) protein were detected in 71% of homogenates of high-grade glioma, but not in the related blood samples. Patients with early tumor progression had lower expression levels of MIA than patients with late progression, and the expression level of MIA was inversely related to time to progression. In addition, MIA expression correlated with a high fiber content of the extracellular matrix, suggesting a role in dissemination as known from malignant melanoma. Expression levels of MIA in homogenates of surgical specimen directly relate to a more benign clinical prognosis in patients with high-grade glioma. While a mechanistic relation has not yet been verified, factors such as a high fiber content of the extracellular matrix may explain this observation. PMID- 15547764 TI - Effect of lung resistance-related protein on the resistance to cisplatin in human ovarian cancer cell lines. AB - The mechanisms of drug-resistance in human ovarian cancer cells have not been entirely clarified. The purpose of this study was to investigate whether LRP is involved in the resistance of ovarian cancer cell lines to cisplatin and its molecular mechanism. Human ovarian cisplatin-resistant cancer cell lines (A2780/DDP and COC1/DDP) and their parental cisplatin-sensitive cell lines (A2780 and COC1), alone or transfected with antisense LRP-specific oligonucleotides (ODN) or sense ODN, were treated with cisplatin to induce differentiation. Expression of LRP was examined by RT-PCR and Western blot analysis. The sensitivities of cells to cisplatin were assessed using sulforhodamine B (SRB) assay and flow cytometry, and the accumulation and efflux of cisplatin in the cells and isolated nuclei were examined by high performance liquid chromatographic (HPLC) assay. The expressions of LRP in A2780/DDP and COC1/DDP cells were higher than those in A2780 and COC1 cells and conferred resistance to cisplatin. Transfection of LRP AsODN into A2780/DDP and COC1/DDP cells down regulated LRP expression and reversed the resistance phenotype. Levels of cisplatin accumulating in cells were increased by LRP-specific AsODN and anti-LRP monoclonal antibody. Isolated nuclei from A2780 and COC1 cells or A2780/DDP and COC1/DDP cells incubated with anti-LRP antibody contained more cisplatin than the nuclei of A2780/DDP and COC1/DDP cells not treated with anti-LRP antibody. Efflux of cisplatin was greater from the nuclei of A2780/DDP and COC1/DDP cells than those of A2780 and COC1 cells, and was inhibited by anti-LRP monoclonal antibody. Thus, LRP was involved in the resistance of ovarian cancer cells to cisplatin and has an important role in the transport of cisplatin both in exocytotic vesicles and between the nucleus and cytoplasm. PMID- 15547765 TI - Lovastatin potentiates antitumor effects of saquinavir against human lymphoma cells. AB - Antiretroviral protease inhibitors have been shown to cause hyperlipidemia, raising concerns for the possibility of cardiovascular complications among HIV infected patients. Statins have been successfully used in the management of hypercholesterolemia and were shown to exert antitumor effects. We report here that lovastatin and saquinavir exert cytostatic/cytotoxic effects against Daudi and Raji human lymphoma cells. Importantly, lovastatin potentiates the antitumor effects of saquinavir against these lymphomas in an additive manner. Addition of statins to antiretroviral treatment might therefore decrease some of its side effects while potentiating the antitumor effectiveness. PMID- 15547766 TI - A matter of life and breath. PMID- 15547767 TI - Total parenteral nutrition induced liver pathology: an autopsy series of 24 newborn cases. AB - Total parenteral nutrition (TPN)-induced liver injury is a common complication in neonates managed with newborn intensive care. In several of these cases, irreversible and even fatal liver damage may develop, with patients dying of liver failure. In spite of multiple studies over several years, the pathogenesis of TPN-induced liver damage remains poorly understood. Clinical data from 24 neonates with clinical history of receiving TPN who died at Yale-New Haven Children's Hospital and had autopsies performed, were collected by medical record review without knowledge of liver pathology findings. Liver histological sections from these patients were evaluated for multiple parameters without knowledge of the clinical course. Continuous data were analyzed by Wilcoxon signed-rank test and Mann-Whitney test, and dichotomous data by Fisher's exact test; P < 0.05 was considered significant. Different histopathological abnormalities with varying degrees of severity were observed. A progression in the severity of histopathological changes in relation to duration of TPN administration (DTPN) was found. While patients with DTPN of < 2 wk had no fibrosis or only mild degrees of fibrosis, patients with more than 6 wk of DTPN developed moderate-to severe fibrosis. Similar results were observed for cholestasis and bile duct proliferation. We did not find significant differences for birth weight, gestational age, occurrence of necrotizing enterocolitis, sepsis, or enteral feedings between the group with normal-to-mild liver changes ( n = 16), and the group with moderate-to-severe liver changes ( n = 8). On the other hand, DTPN was significantly different between these two groups ( P = 0.008). Also, patients small for gestational age ( P = 0.003) and patients with bronchopulmonary dysplasia ( P = 0.001) were more commonly seen in the group with moderate-to severe histopathological findings. Intracellular copper was detected in 12.5% of patients with moderate-to-severe liver changes, and was found in 50% of patients with normal-to-mild liver findings ( P = 0.04). Detection of copper from tissue sections also decreased with DTPN, being observed in 57% of patients with < 2 wk DTPN and in none of the patients with > 12 wk DTPN. Our findings confirm the known significant relationship between the duration of TPN and liver injury. While previously described associations with birth weight, gestational age, enteral feedings, necrotizing enterocolitis, and sepsis were not noted, our study suggests that poor intrauterine growth may be a significant clinical risk factor for TPN-induced liver injury. In addition, our findings suggest that copper may have a protective effect against the development of TPN-induced liver damage. PMID- 15547768 TI - Fetal vascular obstructive lesions: nosology and reproducibility of placental reaction patterns. AB - The purpose of this study was to assemble and test the reliability of a complete set of the placental reaction patterns seen with chronic fetal vascular obstruction in the hope that this might provide a standardized diagnostic framework useful for practicing pathologists. Study cases (14 with fetal vascular obstructive lesions, 6 controls) were reviewed blindly by seven pathologists after agreement on a standard set of diagnostic criteria. Majority vote served as the gold standard and 80% of the 180 diagnoses rendered (9 diagnoses each for 20 cases) were agreed upon by at least six of the seven scores. The sensitivity of individual diagnosis relative to the group consensus averaged 83% (range, 69 100%) and specificity averaged 91% (range, 86-100%). Reproducibility was measured by unweighted kappa-values and interpreted as follows: < 0.2, poor; 0.2-0.6, fair/moderate; > 0.6, substantial. Kappa values for lesions of distal villi were generally superior to those for lesions involving large fetal vessels: avascular villi (0.49), villous stromal-vascular karyorrhexis (0.58), and villitis of unknown etiology (VUE) with stem villitis and avascular villi (0.65) versus large vessel thrombi (any vessel, 0.34; chorionic plate vessel, 0.40) and intimal fibrin cushions (recent, 0.47; remote, 0.78). Reproducibility for a global impression of any villous change consistent with chronic fetal vascular obstruction was substantial (0.63), while that for a more severe subgroup was moderate (0.44). Three points are worthy of emphasis. Our system separately recognizes, but later combines, uniformly avascular villi and villous stromal vascular karyorrhexis as manifestations of the same underlying process. We propose that this combined group of villous lesions be dichotomized with the terms fetal thrombotic vasculopathy or extensive avascular villi (and/or villous stromal-vascular karyorrhexis) being reserved for the group with 15 or more affected terminal villi per section. Scattered foci of avascular villi (and/or villous stromal-vascular karyorrhexis) could be used to describe less severe cases. Finally, we distinguish VUE with stem villitis and avascular villi (obliterative fetal vasculopathy) as a distinct process with substantial perinatal morbidity. PMID- 15547769 TI - Characterization of inflammation in syphilitic villitis and in villitis of unknown etiology. AB - Chronic villitis is a histologic diagnosis that may be either associated with infection, or termed villitis of undetermined etiology (VUE). The lymphocytic infiltrate in VUE has been reported to consist of maternal lymphocytes, but the origin of the lymphocytic infiltrate in infectious villitis has not been identified. The purpose of our study was to compare the maternal vs. fetal origin of the infiltrating lymphocytes in VUE and syphilitic villitis, and to expand the immunophenotypic data provided by previous studies. Paraffin-embedded placentas from four males with VUE and two males with syphilitic villitis were subjected to fluorescence in situ hybridization (FISH) for the X and Y chromosomes. Serial sections were stained with antibodies to CD3, CD4, CD8, CD68, HLA-DR, and CD20. Quantitation of the relative number of cells marking with each antibody was done for four villi in each slide. CD3 lymphocytes predominated in both VUE and syphilitic villitis, with slightly more CD8 cells compared to CD4 cells. CD68 and HLA-DR positive cells were as frequent as CD3 cells, and B-lymphocytes were rare. Maternal cells were the predominant intravillous population in both VUE and syphilitic villitis, and neutrophils in syphilitic villitis were also maternal. These data indicate that the immune response in both syphilitic villitis and VUE is similar, raising the possibility of a similar immunopathogenetic pathway. PMID- 15547770 TI - Postmortem findings and clinicopathological correlation in congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is a severe life-threatening disease, with an incidence of 3 per 10,000 births, that can occur as an isolated defect or in combination with other congenital anomalies. We reviewed the clinical and autopsy reports of 39 subjects with CDH that were autopsied between 1988 and 2001 to determine whether autopsy had an additional value in the detection of malformations in patients with CDH. We compared the clinical data (including echographic results in some patients) concerning congenital anomalies with the autopsy results. Before autopsy, 6 structural cardiac defects, 3 anomalies of the urogenital system, and 3 anomalies of the digestive tract were observed in 10 patients (clinical and echographic results). However, with postmortem examination, only 4 structural cardiac defects were confirmed, 2 cases showed another cardiac anomaly, and 7 new cardiac defects were found. In the urogenital system, 1 anomaly was confirmed, 1 was not confirmed, and 1 showed another malformation. In addition, in 7 patients new urogenital malformations were found after autopsy. In the digestive tract, all 3 malformations were confirmed, but we found 3 new malformations after postmortem examination. All clinically established dysmorphic features and anomalies of the skeletal system and central nervous system were confirmed by autopsy, and no additional malformations were found. We concluded that postmortem examination has an important additional role in the detection of structural cardiac defects and malformations of the urogenital system and digestive tract in children with CDH. PMID- 15547771 TI - Pathology of human metapneumovirus infection: insights into the pathogenesis of a newly identified respiratory virus. AB - Human metapneumovirus (hMPV) is a recently discovered human virus that causes significant respiratory infections. Pathologic features of hMPV infection have not been described. A total of 1257 pediatric respiratory samples submitted for routine clinical virologic testing were additionally tested for hMPV by reverse transcriptase polymerase chain reaction (PCR). Pathology specimens, available in 6 of 53 hMPV-positive patients, were examined by light and electron microscopy and included 6 bronchoalveolar lavage (BAL) and 3 lung biopsy specimens from 6 patients (3 girls and 3 boys) ranging in age from 1 to 16 years. BAL from three patients performed within 4 days of the positive hMPV assay showed epithelial degenerative changes and eosinophilic cytoplasmic inclusions within epithelial cells, multinucleate giant cells, and histiocytes. Inclusions were not seen in three patients with BAL performed = 1 month from the time of their positive assay. Lung biopsy, performed in three patients, all = 1 month from the time of their positive assay, showed chronic airway inflammation and intraalveolar foamy and hemosiderin-laden macrophages; all three patients had an underlying pulmonary/systemic disorder. Our findings delineate the clinicopathologic features in hMPV-infected patients undergoing anatomic sampling, which may provide diagnostic guidance to a practicing pathologist. Further, they contribute toward understanding the pathogenesis of hMPV infection. PMID- 15547772 TI - Lymphocytic infiltration in pediatric thyroid carcinomas. AB - Lymphocytic thyroiditis has been associated with an increase in the incidence of thyroid papillary carcinoma in some reports, mostly series of both adults and children. Relatively little is written about thyroiditis and follicular carcinomas. We have seen several cases of pediatric follicular thyroid carcinomas, that had an associated lymphocytic infiltrate, which led us to examine all primary malignant thyroid neoplasms in our surgical files from 1984 through 2000 to examine this relationship. We also investigated the nature of the lymphocytic infiltrate with routine immunohistochemistry. Ten patients (five male, five female, ages 4.5-21 years of age) had a thyroid carcinoma resection, six (three males and three females) with papillary carcinoma and four patients (two males and two females) with low-grade follicular carcinoma. Seven samples (one male had two cases with tumor) from patients who had a papillary carcinoma resection with tissue blocks available were identified (one patient had slides but no blocks), as were all four patients with a follicular carcinoma. The thyroid of all patients with a follicular carcinoma contained a lymphocytic infiltrate; only four of the seven papillary carcinoma samples had an associated lymphoid infiltrate. In all cases with a lymphoid infiltrate, the infiltrate was present in both lobes (both adjacent and separate from the tumor). B lymphocytes were present in the lymphoid infiltrate of three of four patients with follicular carcinomas and in 1 of 3 cases of papillary carcinomas. T cells were dispersed throughout all the tumors with lymphoid infiltrates. We conclude that pediatric follicular carcinomas have an associated lymphocytic infiltrate in the tumor and/or adjacent thyroid, more commonly than papillary carcinomas. PMID- 15547773 TI - C-kit expression and mutational analysis in medulloblastoma. AB - The proto-oncogene c-kit is a receptor tyrosine kinase recognized to initiate essential signal transduction pathways that transmit biological signals for cellular proliferation, differentiation, and metastasis. Aberrant expression or mutation of c-kit has been shown to be involved in the pathogenesis of many cancers. Studies using imatinib mesylate (STI 571, Gleevec, Novartis, East Hannover, NJ, USA), an inhibitor of the tyrosine kinases brc-abl, c-kit, and PDGFR, have shown significant response in patients with chronic myelogenous leukemia and gastrointestinal stromal tumor. With the aim of identifying additional groups of tumors that may use the stem cell factor/c-kit pathway and, secondarily, may be responsive to imatinib mesylate treatment, we looked at the expression of c-kit in medulloblastoma. Medulloblastoma, a highly invasive primitive neuroectodermal tumor of the cerebellum, is the most common, malignant central nervous system tumor of childhood. Histologic features of medulloblastoma have failed to provide an accurate prediction of the clinical-biological behavior of these tumors. Characterizing the genetic events that play a role in the biology of these tumors may allow for molecular sub-typing and could lead to the development of novel therapeutic strategies. This study evaluated c-kit expression and mutational status in 10 medulloblastoma tumor samples. All 10 medulloblastoma tumors expressed c-kit by reverse transcriptase-polymerase chain reaction and 9 by immunohistochemical analysis. All tumor samples were screened for mutations in exons 9, 11, and 13 of the c-kit gene by direct sequencing. No sequence abnormalities were detected in these exons. These experiments lead us to the conclusion that c-kit activation in medulloblastoma is independent of mutation. PMID- 15547774 TI - Reference values for valve circumferences and ventricular wall thicknesses of fetal and neonatal hearts. AB - To evaluate valvular stenosis, cardiac dilation, and/or cardiac hypertrophy, measurements of valve circumference and ventricular wall thickness are of importance. To establish reference values in fetuses and neonates, we reviewed pathology reports at Women and Infants Hospital from 1978 through 2002 and found measurements in 776 cases that were suitable for analysis. Gestational ages (GA) ranged from 15 to 42 wk. The tabulated data include the mean, standard deviation, and 10th and 90th percentile values for foot length, body weight, body length, heart weight, valve measurements, and ventricular wall thicknesses for each week of GA. In cases in which clinical dating is not reliable, we estimated the GA by the mean value nearest that of the observed foot length. All linear measurements increased in a linear fashion throughout the second and third trimesters of development. The circumferences of cardiac valves at all ages, in descending order of magnitude, are: tricuspid, mitral, pulmonary, and aortic. Mean left ventricular (LV) wall thickness is greater than mean right ventricular (RV) wall thickness throughout gestation. The tables offer a means of determining valvular stenosis, or cardiac dilation and/or hypertrophy, based on various gestational ages. PMID- 15547775 TI - Alpha-synuclein expression in the developing human brain. AB - Alpha (alpha)-synuclein is a presynaptic protein, abnormal expression of which has been associated with neurodegenerative and neoplastic diseases. It is abundant in the developing vertebrate central nervous system (CNS), but less is known about its developmental expression in the human CNS. Immunohistochemical expression of alpha-synuclein was studied in 39 fetal, perinatal, pediatric, and adolescent brains. Perikaryal expression of alpha-synuclein is observed as early as 11-wk gestation in the cortical plate. Several discrete neuronal groups in the hippocampus, basal ganglia, and brain stem express perikaryal alpha-synuclein by 20-wk gestation, persisting through the first few years of life. In the cerebellum, alpha-synuclein is present by 21-wk gestation and persists into adult life as a coarse granular neuropil reaction product in the internal granular layer, and as a diffuse neuropil "blush" in the molecular layer. The germinal matrix, glia, endothelial cells, external granular layer, Pukinje cells, and dentate neurons are consistently negative for alpha-synuclein. We conclude that alpha-synuclein is expressed very early in human gestation, and that its distribution and temporal sequence of expression varies in discrete neuronal groups. Perikaryal alpha-synuclein starts disappearing from the neuronal cytosol in early childhood, and only the neuropil retains immunoreactivity into adulthood. The reappearance of alpha-synuclein in the adult neuronal cytosol in certain disease processes may represent reemergence of cues from an earlier developmental stage as part of a stress response. PMID- 15547776 TI - Nasal chondromesenchymal hamartoma: report of a case and review of the literature. AB - The sinonasal region is the site of several hamartomatous lesions, the majority of which are mesenchymal, with vascular hamartomas predominating. The occurrence of hamartomas in the nasal cavity of infants and children is especially rare. Nasal chondromesenchymal hamartoma (NCMH) is a rare lesion of the intranasal sinuses generally diagnosed in the newborn period, with the eldest reported patient presenting at 16 years of age. This neoplasm is composed of mesenchymal stromal and chondroid tissue in varying proportions. It is felt to be analogous to the mesenchymal hamartoma of the chest wall, a lesion of similar histology generally involving the ribs and chest wall of neonates. To the best of our knowledge, only 14 cases of NCMH have been reported to date. We report a case of NCMH in an 11-year-old boy. PMID- 15547777 TI - Pediatric hepatic angiosarcoma: case report and review of the literature. AB - Pediatric hepatic angiosarcoma (PHAS) is a rare tumor, which usually presents as a rapid enlargement of the liver. To date, surgery, chemotherapy, and radiotherapy have not improved the poor prognosis of PHAS with only three survivors reported. The histology of PHAS is distinct from adult angiosarcoma, because PHAS displays hypercellular whorls of sarcomatous cells, or "kaposiform" spindle cells, in addition to the general features of angiosarcoma. We report a case of PHAS that was treated with vascular ablation, chemotherapy, and liver transplantation. Lung metastases occurred 14 months posttransplant. PMID- 15547778 TI - Transient posttransplant graft-versus-host lymphadenopathy. AB - There is sparse information in humans on graft-versus-host (GVH) lymphadenopathy. A 15-month-old male received a liver and small bowel transplant for short bowel after gastroschisis. At 21 days he developed a GVH-like skin rash. Flow cytometry demonstrated 16.1% circulating donor cells. Polymerase chain reaction for Epstein Barr virus was negative. Two months later, the rash recurred with diffuse lymphadenopathy. Lymph node biopsy showed effaced architecture without visible follicles, large numbers of CD79a(+) immunoblasts interspersed with smaller CD3(+) and CD8(+) cells, and prominent dendritic cell hyperplasia. Human herpes virus 8, cytomegalovirus, and EBER-1 probes were negative, as was polymerase chain reaction for human herpes virus 6. Allograft intestinal biopsies on days 10 and 24 had a similar infiltrate. The features appeared to be those of lymphocytes trafficking between the graft and host with a mixed lymphocyte reaction in situ, a GVH-type reaction without tissue damage. The reaction was self-limiting in the intestinal graft, and the lymphadenopathy resolved with some decrease in immunosupression. Circulating donor cells fell to 2.5% by day 62, and the child has been rejection free on low-dose immunosuppression. PMID- 15547779 TI - Primary subcutaneous primitive neuroectodermal tumor with aggressive behavior and an unusual karyotype: case report. AB - Primitive neuroectodermal tumor/Ewing sarcoma (PNET/ES) rarely occurs in the skin and subcutaneous tissues. We present a case of a 16-year-old girl with primary cutaneous and subcutaneous PNET/ES of the abdominal wall. Despite wide local excision and chemotherapy, she rapidly developed cranial bone and brain metastases, followed by lung and skeletal metastases, and died shortly thereafter. The recurrent tumor exhibited light microscopic features of a small, round, blue cell tumor with intracytoplasmic glycogen. Immunohistochemical analysis showed positivity for CD99, CD56, S100, and glial fibrillary acid protein, and ultrastructural features included cytoplasmic glycogen and focal complex interdigitating synaptic junction-like cytoplasmic folds. Cytogenetic analysis of the relapsed tumor showed a complex karyotype: 47,XX,i(1)(q10), der(4)t(4;19) (q33 approximately q35;q13.1), + 8,t(15;17)(q24;p11.2 approximately p12),der(19)t (19;20)(q13.1;p11.2),der(22)t(20;22)(q13;q13). Cytogenetic, interphase fluorescence in situ hybridization, and molecular genetic analyses failed to show t(11:22) (q24;q12) or abnormalities of chromosome region 22q12. The clinical behavior and atypical and complex cytogenetic abnormalities exhibited by the tumor in this patient are unusual and represent the most aggressive end of the clinical spectrum of cutaneous and subcutaneous PNET/ES. PMID- 15547780 TI - Dysmorphic villi mimicking partial mole in a case with del(18)(q21). AB - In this article, we report the histopathologic findings in a placenta from an early second trimester abortion. The placental villi showed prominent scalloping with many intravillous trophoblastic pseudoinclusions and mild trophoblastic hyperplasia, mimicking the morphology of partial hydatidiform mole. The placental karyotype was 46,XY,del(18)(q21). These histopathologic changes have been previously described in numerical chromosomal aberrations like triploidy, tetraploidy, and trisomies, but not in structural chromosomal abnormalities. PMID- 15547781 TI - Editorial. PMID- 15547782 TI - Sigma receptors and sigma ligands: background to a pharmacological enigma. AB - Although it is now well established that the sigma receptors are distinct from neurotransmitter, neuropeptide and steroid receptors, their precise physiological role, and possible involvement in the pathology of neurological and psychiatric disorders, remains an enigma. The purpose of this short review is to provide a basis upon which to explain the pharmacological properties of such a diverse group of drugs as the antidepressants, antipsychotics and neuroprotective agents. However, a greater understanding of the importance of the sigma receptors will depend on the development of more potent and selective sigma agonists and antagonists if the therapeutic potential of the sigma receptors is to be fully realised. PMID- 15547783 TI - Neurosteroids and sigma1 receptors, biochemical and behavioral relevance. AB - The sigma(1) receptor is a 223 amino acid protein sharing no homology with other mammalian protein. It is an intracellular protein present on the endoplasmic reticulum membrane, which can translocates to other organelles and plasma membranes after activation. Activation of the sigma(1) receptor results in modulation of calcium mobilization from inositol trisphosphate receptor-gated intracellular pools and, at the plasma membrane, in modulation of several neurotransmitter responses. Behaviorally, sigma(1) receptors are involved in learning and memory, response to stress and depression, psychostimulant-induced sensitization, vulnerability to addiction and pain perception. Numerous synthetic compounds bind to sigma(1) receptor, playing the role of activator/agonist or blocker/antagonist, and these include benzomorphans, neuroleptics, antidepressants, cocaine, peptides related to neuropeptide Y or calcitonin gene related peptide. It is also the case of neuro(active)steroids, i. e., circulating neuroactive steroids and neurosteroids synthesized de novo by the brain, which appear as the most important endogenous modulators of sigma(1) receptor. Pregnenolone and dehydroepiandrosterone act as sigma(1) receptor agonists and progesterone is a potent antagonist. The present paper will review the molecular and biochemical features concerning the sigma(1) receptor and focus on the recent studies examining the impact of the neuro(active)steroid/sigma(1) receptor interaction on the antidepressant activity of sigma(1) receptor agonists in the context of neurodegenerative diseases. PMID- 15547784 TI - Behavioral pharmacology of sigma-ligands. AB - Sigma (sigma) receptors, first defined as a subclass of opioid receptors, later confounded with the high affinity phencyclidine (PCP) binding sites, now are regarded as unique binding sites, distinct from opiate and PCP receptors, and related to higher brain function. The investigation of functional significance of sigma receptors in the brain has been hampered for many years by relative lack of specific tool drugs and by the unavailability of their coherent classification into postulated agonists and antagonists. However, a potential involvement of sigma receptors in psychotic disorders was first suggested soon after their discovery. The sigma receptors are classified into two subtypes, sigma (1) and sigma (2) receptors, of which the first was recently cloned from rodent and human tissues while the second has not yet been fully characterized. Although the precise mechanism of the functional response of these receptors is still uncertain, it is accepted that sigma receptors can modulate a number of central neurotransmitter systems, including noradrenergic, glutamatergic and dopaminergic ones. The sigma receptors have been postulated to be involved in numerous pharmacological and physiological functions, including motor disorders, psychotic disorders, neuroprotective mechanisms. In the last years, a number of compounds with a high affinity and selectivity for sigma binding sites have been discovered and investigated for their therapeutic potential. In this review, we try to summarize the behavioral effects of sigma receptor ligands that have been described, and their activity in animal models related to some brain disorders, especially schizophrenia and affective disorders. PMID- 15547785 TI - Neuropharmacology of the anxiolytic drug opipramol, a sigma site ligand. AB - Although opipramol is structurally related to imipramine, it does not represent a tricyclic antidepressant drug as it does not inhibit the neuronal uptake of norepinephrine and/or serotonin. Unlike imipramine it is a rather potent sigma ligand with modest subclass selectivity which is similar in vitro as well as ex vivo. Opipramol is active in several behavioural paradigms indicative of anxiolytic properties at doses (1-10 mg/kg), which are also needed to occupy sigma binding sites. Somewhat higher doses (10-20 mg/kg) are needed for "antidepressant like" effects. The data allow the conclusion that interaction with sigma sites is involved in the anxiolytic and antidepressant effects of opipramol albeit a contribution of its weaker D (2)-antagonistic and 5-HT2 antagonistic properties cannot be totally be excluded. PMID- 15547786 TI - Involvement of the sigma1 receptor in the appetitive effects of cocaine. AB - Cocaine induces in the brain a drastic plasticity affecting numerous neurotransmission systems. In turn, the drug provokes a complex pattern of behavioral responses, including locomotor hyperactivity, stereotyped behaviors, sensitization, and appetitive and reinforcing properties that rapidly generate the addictive process. The involvement of sigma (1) receptors in cocaine effects has been suggested initially through the observation that sigma (1) antagonists could attenuate several acute effects induced by the drug, such as hyperlocomotion, stereotypies, convulsions and lethality. We will describe thereafter the recent results showing that activation of the sigma (1) receptor is also involved in the appetitive properties of cocaine, as measured using place preference conditioning in mice. Using selective sigma (1) antagonists or an in vivo antisense strategy, the role of the sigma (1) receptor in acquisition, expression and reactivation of conditioned place preference was demonstrated. The observation that repeated administration of cocaine rapidly provokes overexpression of the sigma (1) receptor outlines its major role in these first psychological steps of addictive processes. The physiological interaction between cocaine and sigma (1) receptors is detailed and the possibility that effective therapeutical strategies could target the sigma (1) receptor is considered. PMID- 15547787 TI - A perspective on the new mechanism of antidepressants: neuritogenesis through sigma-1 receptors. AB - Sigma receptors were first described as one of the opiate receptor subtypes. Now it is well established that sigma receptors, existing as subtypes sigma-1 and sigma-2, are unique non-opioid receptors which are implicated in higher-ordered brain functions. Sigma-1 receptors have high to moderate affinities for (+)benzomorphans and also many psychotrophic drugs and neurosteroids. Sigma-1 receptor agonists and certain neurosteroids such as dehydroepiandrosterone sulfate (DHEA-S) have antidepressant-like effects in animal behavioral models of depression. The antidepressant-like effect induced by sigma-1 receptor agonists may involve intracellular Ca (2+) mobilization such that sigma-1 receptor agonists modulate Ca (2+) release from endoplasmic reticulum (ER) in a cytoskeletal protein-dependent manner. In addition, growth factor-induced neurite outgrowth is mediated through sigma-1 receptors, suggesting a role of antidepressants in neuroplasticity. Igmesine (JO1783), OPC-14 523 and SA4503, have recently been developed as sigma-1 agonists and are found to have antidepressant-like activity perhaps with fewer side effects. This article reviews the new potential use of sigma-1 receptor ligands in the treatment of mood disorder. PMID- 15547788 TI - Clinical trials with sigma ligands. AB - So far, sigma-ligands have been investigated for several indications in human studies in functional diarrhea as a model of somatoform disorder (igmesine), depression (igmesine, opipramol), anxiety (opipramol and--in animal models- siramisine), schizophrenia (panamasine, SL 82.0715, rimcazole, DuP 734, BMY 14 802), and somatoform disorders (opipramol). Results for schizophrenia failed to be clear cut and so investigations have apparently stopped for the time being. The Sigma-1-selective igmesine (200 mg) showed good results in a phase-1-model of functional diarrhea and some promising results in depressed patients. However, further development has been stopped due to marketing reasons, which is also true for siramasine, a selective sigma-2-ligand with anxiolytic properties. Opipramol, which, apart from a sigma-1- and 2-receptor liability, also possesses histamine H(1)-antagonistic properties in connection with lower affinities for D(2) and 5 HT(2A) showed broad efficacy in generalized anxiety disorder and somatoform disorders. The receptor profile of opipramol and the results of studies of the selective sigma site ligands siramisine and igmesine suggest that opipramol acts pharmacologically and clinically via sigma receptors. PMID- 15547790 TI - The greatest step in vertebrate history: a paleobiological review of the fish tetrapod transition. AB - Recent discoveries of previously unknown fossil forms have dramatically transformed understanding of many aspects of the fish-tetrapod transition. Newer paleobiological approaches have also contributed to changed views of which animals were involved and when, where, and how the transition occurred. This review summarizes major advances made and reevaluates alternative interpretations of important parts of the evidence. We begin with general issues and concepts, including limitations of the Paleozoic fossil record. We summarize important features of paleoclimates, paleoenvironments, paleobiogeography, and taphonomy. We then review the history of Devonian tetrapods and their closest stem group ancestors within the sarcopterygian fishes. It is now widely accepted that the first tetrapods arose from advanced tetrapodomorph stock (the elpistostegalids) in the Late Devonian, probably in Euramerica. However, truly terrestrial forms did not emerge until much later, in geographically far-flung regions, in the Lower Carboniferous. The complete transition occurred over about 25 million years; definitive emergences onto land took place during the most recent 5 million years. The sequence of character acquisition during the transition can be seen as a five-step process involving: (1) higher osteichthyan (tetrapodomorph) diversification in the Middle Devonian (beginning about 380 million years ago [mya]), (2) the emergence of "prototetrapods" (e.g., Elginerpeton) in the Frasnian stage (about 372 mya), (3) the appearance of aquatic tetrapods (e.g., Acanthostega) sometime in the early to mid-Famennian (about 360 mya), (4) the appearance of "eutetrapods" (e.g., Tulerpeton) at the very end of the Devonian period (about 358 mya), and (5) the first truly terrestrial tetrapods (e.g., Pederpes) in the Lower Carboniferous (about 340 mya). We discuss each of these steps with respect to inferred functional utility of acquired character sets. Dissociated heterochrony is seen as the most likely process for the evolutionarily rapid morphological transformations required. Developmental biological processes, including paedomorphosis, played important roles. We conclude with a discussion of phylogenetic interpretations of the evidence. PMID- 15547791 TI - Breathing air in air: in what ways might extant amphibious fish biology relate to prevailing concepts about early tetrapods, the evolution of vertebrate air breathing, and the vertebrate land transition? AB - The air-breathing fishes have heuristic importance as possible models for the Paleozoic evolution of vertebrate air breathing and the transition to land. A recent hypothesis about this transition suggests that the diverse assemblage of marine amphibious fishes occurring primarily in tropical, high intertidal zone habitats are analogs of early tetrapods and that the intertidal zone, not tropical freshwater lowlands, was the springboard habitat for the Devonian land transition by vertebrates. Here we argue that selection pressures imposed by life in the intertidal zone are insufficient to have resulted in the requisite aerial respiratory capacity or the degree of separation from water required for the vertebrate land transition. The extant marine amphibious fishes, which occur mainly on rocky shores or mudflats, have reached the limit of their niche expansion onto land and remain tied to water by respiratory structures that are less efficient in air and more vulnerable to desiccation than lungs. We further argue that evolutionary contingencies actuated by the Devonian origin of the tetrapods marked a critical point of divergence for a way of life in which selection pressures would operate on the physiology, morphology, and natural history of the different vertebrate groups. While chronically hypoxic and shallow water conditions in the habitats of some primitive bony fishes and some amphibians appear similar to the conditions that prevailed in the Devonian, markedly different selection pressures have operated on other amphibians and bony fishes over the 300 million years since the vertebrate land transition. For example, both egg development and larval metamorphosis in extant amphibians are geared mainly toward compensating for the uncertainty of habitat water quality or even the absence of water by minimizing the time required to develop there. In contrast, reproduction by most intertidal (and amphibious) fishes, all of which are teleosts, remains dependent on a planktonic larval phase and is characterized by specializations (brooding) that minimize overdispersal and maximize recruitment back to the littoral habitat. PMID- 15547792 TI - The origin and evolution of the surfactant system in fish: insights into the evolution of lungs and swim bladders. AB - Several times throughout their radiation fish have evolved either lungs or swim bladders as gas-holding structures. Lungs and swim bladders have different ontogenetic origins and can be used either for buoyancy or as an accessory respiratory organ. Therefore, the presence of air-filled bladders or lungs in different groups of fishes is an example of convergent evolution. We propose that air breathing could not occur without the presence of a surfactant system and suggest that this system may have originated in epithelial cells lining the pharynx. Here we present new data on the surfactant system in swim bladders of three teleost fish (the air-breathing pirarucu Arapaima gigas and tarpon Megalops cyprinoides and the non-air-breathing New Zealand snapper Pagrus auratus). We determined the presence of surfactant using biochemical, biophysical, and morphological analyses and determined homology using immunohistochemical analysis of the surfactant proteins (SPs). We relate the presence and structure of the surfactant system to those previously described in the swim bladders of another teleost, the goldfish, and those of the air-breathing organs of the other members of the Osteichthyes, the more primitive air-breathing Actinopterygii and the Sarcopterygii. Snapper and tarpon swim bladders are lined with squamous and cuboidal epithelial cells, respectively, containing membrane-bound lamellar bodies. Phosphatidylcholine dominates the phospholipid (PL) profile of lavage material from all fish analyzed to date. The presence of the characteristic surfactant lipids in pirarucu and tarpon, lamellar bodies in tarpon and snapper, SP-B in tarpon and pirarucu lavage, and SPs (A, B, and D) in swim bladder tissue of the tarpon provide strong evidence that the surfactant system of teleosts is homologous with that of other fish and of tetrapods. This study is the first demonstration of the presence of SP-D in the air-breathing organs of nonmammalian species and SP-B in actinopterygian fishes. The extremely high cholesterol/disaturated PL and cholesterol/PL ratios of surfactant extracted from tarpon and pirarucu bladders and the poor surface activity of tarpon surfactant are characteristics of the surfactant system in other fishes. Despite the paraphyletic phylogeny of the Osteichthyes, their surfactant is uniform in composition and may represent the vertebrate protosurfactant. PMID- 15547793 TI - Beach-spawning fishes, terrestrial eggs, and air breathing. AB - Many fishes have independently evolved beach spawning with oviposition at the water's edge. These include intertidal, subtidal, and estuarine, as well as a few freshwater, species. Their spectacular reproductive behavior at the boundary of water and land has focused attention on adults, but they emerge either briefly or not at all. The need for air breathing is more apparent in the eggs, and the reasons for emergence are more applicable to eggs than to the adults of most beach-spawning fishes. There is little evidence of air breathing in the adults, unless they are regularly emerged at other times as well. Conversely, eggs metabolize in air and show substantial emergence tolerance. We consider beach spawning a form of parental care in fishes. The adults place eggs so they will be emerged into air during part or all of incubation, providing increased temperatures, oxygen availability, and protection. Beach spawning provides habitat segregation at different points in the life history, with air emergence early in the life cycle and a return to water at hatching. The parents take great risks to spawn at the water's edge to give their offspring the most advantageous beginning in life. PMID- 15547794 TI - Partitioning of respiration between the gills and air-breathing organ in response to aquatic hypoxia and exercise in the pacific tarpon, Megalops cyprinoides. AB - The evolution of air-breathing organs (ABOs) is associated not only with hypoxic environments but also with activity. This investigation examines the effects of hypoxia and exercise on the partitioning of aquatic and aerial oxygen uptake in the Pacific tarpon. The two-species cosmopolitan genus Megalops is unique among teleosts in using swim bladder ABOs in the pelagic marine environment. Small fish (58-620 g) were swum at two sustainable speeds in a circulating flume respirometer in which dissolved oxygen was controlled. For fish swimming at 0.11 m s(-1) in normoxia (Po2 = 21 kPa), there was practically no air breathing, and gill oxygen uptake was 1.53 mL kg(-0.67) min(-1). Air breathing occurred at 0.5 breaths min(-1) in hypoxia (8 kPa) at this speed, when the gills and ABOs accounted for 0.71 and 0.57 mL kg(-0.67) min(-1), respectively. At 0.22 m s(-1) in normoxia, breathing occurred at 0.1 breaths min(-1), and gill and ABO oxygen uptake were 2.08 and 0.08 mL kg(-0.67) min(-1), respectively. In hypoxia and 0.22 m s(-1), breathing increased to 0.6 breaths min(-1), and gill and ABO oxygen uptake were 1.39 and 1.28 mL kg(-0.67) min(-1), respectively. Aquatic hypoxia was therefore the primary stimulus for air breathing under the limited conditions of this study, but exercise augmented oxygen uptake by the ABOs, particularly in hypoxic water. PMID- 15547795 TI - Five tropical air-breathing fishes, six different strategies to defend against ammonia toxicity on land. AB - Most tropical fishes are ammonotelic, producing ammonia and excreting it as NH3 by diffusion across the branchial epithelia. Hence, those air-breathing tropical fishes that survive on land briefly or for an extended period would have difficulties in excreting ammonia when out of water. Ammonia is toxic, but some of these air-breathing fishes adopt special biochemical adaptations to ameliorate the toxicity of endogenous ammonia accumulating in the body. The amphibious mudskipper Periophthalmodon schlosseri, which is very active on land, reduces ammonia production by suppressing amino acid catabolism (strategy 1) during aerial exposure. It can also undergo partial amino acid catabolism, leading to the accumulation of alanine (strategy 2) to support locomotory activities on land. In this case, alanine formation is not an ammonia detoxification process but reduces the production of endogenous ammonia. The snakehead Channa asiatica, which exhibits moderate activities on land although not truly amphibious, accumulates both alanine and glutamine in the muscle, with alanine accounting for 80% of the deficit in reduction in ammonia excretion during air exposure. Unlike P. schlosseri, C. asiatica apparently cannot reduce the rates of protein and amino acid catabolism and is incapable of utilizing partial amino acid catabolism to support locomotory activities on land. Unlike alanine formation, glutamine synthesis (strategy 3) represents an ammonia detoxification mechanism that, in effect, removes the accumulating ammonia. The four-eyed sleeper Bostrichyths sinensis, which remains motionless during aerial exposure, detoxifies endogenous ammonia to glutamine for storage. The slender African lungfish Protopterus dolloi, which can aestivate on land on a mucus cocoon, has an active ornithine urea cycle and converts endogenous ammonia to urea (strategy 4) for both storage and subsequent excretion. Production of urea and glutamine are energetically expensive and appear to be adopted by fishes that remain relatively inactive on land. The Oriental weatherloach Misgurnus anguillicaudatus, which actively burrows into soft mud during drought, manipulates the pH of the body surface to facilitate NH3 volatilization (strategy 5) and develops high ammonia tolerance at the cellular and subcellular levels (strategy 6) during aerial exposure. Hence, with regard to excretory nitrogen metabolism, modern tropical air-breathing fishes exhibit a variety of strategies to survive on land, and they represent a spectrum of specimens through which we may examine various biochemical adaptations that would have facilitated the invasion of the terrestrial habitat by fishes during evolution. PMID- 15547796 TI - Air breathing and ammonia excretion in the giant mudskipper, Periophthalmodon schlosseri. AB - The giant mudskipper, Periophthalmodon schlosseri, is an amphibious, obligate, air-breathing teleost fish. It uses its buccal cavity for air breathing and for taking and holding large gulps of air. These fish live in mud burrows at the top of the intertidal zone of mangrove mudflats; the burrow water may be hypoxic and hypercapnic and have high ammonia levels. The buccal epithelium is highly vascularized, with small diffusion distances between air and blood. The gill epithelium is densely packed with mitochondria-rich cells. Periophthalmodon schlosseri can maintain tissue ammonia levels in the face of high ammonia concentrations in the water. This is probably achieved by active ammonium ion transport across the mitochondria-rich cells via an apical Na/H+(NH4+) exchanger and a basolateral Na/K+(NH4+) ATPase. When exposed to air, the animal reduces ammonia production, but there is some increase in tissue ammonia levels after 24 h. There is no detoxification by increased production of glutamine or urea, but there is partial amino acid catabolism, leading to the accumulation of alanine. CO2 production and proton excretion cause acidification of the burrow water to reduce ammonia toxicity. The skin has high levels of cholesterol and saturated fatty acids decreasing membrane fluidity and gas, and therefore ammonia, permeability. Exposure to elevated environmental ammonia further decreases membrane permeability. Acidification of the environment and having a skin with a low NH3 permeability reduces ammonia influx, so that the fish can maintain tissue ammonia levels by active ammonium ion excretion, even in water containing high levels of ammonia. PMID- 15547797 TI - Digestive enzyme activities in herbivorous and carnivorous prickleback fishes (Teleostei: Stichaeidae): ontogenetic, dietary, and phylogenetic effects. AB - We measured the activities of eight digestive enzymes in four species of herbivorous and carnivorous prickleback fishes and determined the effects of ontogeny, diet, and phylogeny on these enzyme activities. Of the four species, Cebidichthys violaceus and Xiphister mucosus shift to a more herbivorous diet as they grow (> or =45 mm SL [standard length]), whereas Xiphister atropurpureus and Anoplarchus purpurescens remain carnivores throughout life. Digestive enzyme activities of small (30-40 mm SL) carnivorous juveniles were compared with those of larger (60-75 mm SL) wild-caught juveniles that had consumed a natural diet and larger (60-75 mm SL) juveniles raised on a high-protein animal diet. Cebidichthys violaceus and both species of Xiphister showed ontogenetic changes in digestive enzyme activities, whereas A. purpurescens did not. Despite dietary differences between X. atropurpureus and X. mucosus, these sister taxa displayed the most similar digestive enzyme activities from ontogenetic and dietary perspectives (high alpha-amylase and lipase and low trypsin and aminopeptidase activities), and both were more similar to C. violaceus, a member of the same largely herbivorous clade, than either was to A. purpurescens, a member of an adjacent, carnivorous clade. The results support the hypothesis that phylogeny influences digestive enzyme activities in these fishes. Anoplarchus purpurescens, a carnivore with a diverse diet, showed great plasticity in enzyme activity, especially trypsin and aminopeptidase, which were elevated in this species to the highest level among the four species after consuming the high-protein diet. These results support the hypothesis that fishes with relatively broad diets can modulate digestive enzyme activities in response to changes in dietary composition. PMID- 15547798 TI - Phenotypic flexibility in a novel thermal environment: phylogenetic inertia in thermogenic capacity and evolutionary adaptation in organ size. AB - The goal of our work was to understand the role of a novel thermal environment in shaping the phenotypic expression of thermogenic capacity and organ size. To examine this we compared two populations of the South American rodent Phyllotis darwini from different altitudes (Andean and valley populations), taking advantage of the fact that this genus originated at high altitude in the Andean plateau. DNA mitochondrial analysis showed that the two populations were separated and then experienced different thermal regimens for at least the last 450,000 yr. We expected the two populations of P. darwini to present more metabolic and organ size similarities if phylogenetic inertia had been an important factor. In this sense, phylogenetic inertia means that the valley population would retain evolutionary adaptations of high altitude: a greater phenotypic flexibility in both physiological and morphological traits. In general, our results indicate that the actual thermogenic capacities (magnitude and flexibility) of the valley population are a consequence of phylogenetic inertia. On the other hand, results for organ size (magnitude and flexibility) could suggest that this population would have adapted to the less seasonal central valley. PMID- 15547799 TI - Endothermy in African platypleurine cicadas: the influence of body size and habitat (Hemiptera: Cicadidae). AB - The platypleurine cicadas have a wide distribution across Africa and southern Asia. We investigate endothermy as a thermoregulatory strategy in 11 South African species from five genera, with comparisons to the lone ectothermic platypleurine we found, in an attempt to ascertain any influence that habitat and/or body size have on the expression of endothermy in the platypleurine cicadas. Field measurements of body temperature (T(b)) show that these animals regulate T(b) through endogenous heat production. Heat production in the laboratory elevated T(b) to the same range as in animals active in the field. Maximum T(b) measured during calling activity when there was no access to solar radiation ranged from 13.2 degrees to 22.3 degrees C above ambient temperature in the five species measured. The mean T(b) during activity without access to solar radiation did not differ from the mean T(b) during diurnal activity. All platypleurines exhibit a unique behavior for cicadas while warming endogenously, a temperature-dependent telescoping pulsation of the abdomen that probably functions in ventilation. Platypleurines generally call from trunks and branches within the canopy and appear to rely on endothermy even when the sun is available to elevate T(b), in contrast to the facultative endothermy exhibited by New World endothermic species. The two exceptions to this generalization we found within the platypleurines are Platypleura wahlbergi and Albanycada albigera, which were the smallest species studied. The small size of P. wahlbergi appears to have altered their thermoregulatory strategy to one of facultative endothermy, whereby they use the sun when it is available to facilitate increases in T(b). Albanycada albigera is the only ectothermic platypleurine we found. The habitat and host plant association of A. albigera appear to have influenced the choice of ectothermy as a thermoregulatory strategy, as the species possesses the metabolic machinery to elevate to the T(b) range observed in the endothermic species. Therefore, size and habitat appear to influence the expression of thermoregulatory strategies in African platypleurine cicadas. PMID- 15547800 TI - Strategies adopted by the mudskipper Boleophthalmus boddaerti to survive sulfide exposure in normoxia or hypoxia. AB - The effects of sulfide on the energy metabolism of Boleophthalmus boddaerti in normoxia and hypoxia were examined. The 24-, 48-, and 96-h LC50 values of sulfide for B. boddaerti with body weight ranging from 11.6 to 14.2 g were 0.786, 0.567, and 0.467 mM, respectively. The tolerance of B. boddaerti to sulfide was not due to the presence of a sulfide-insensitive cytochrome c oxidase. There was no accumulation of lactate in the muscle and liver of specimens exposed to sulfide in normoxia. In addition, the levels of ATP, AMP, and energy charge in both the muscle and the liver were unaffected. These results indicate that B. boddaerti was able to sustain the energy supply required for its metabolic needs via mainly aerobic respiration when exposed to sulfide (up to 0.4 mM) in normoxia. Exposure of B. boddaerti simultaneously to hypoxia and 0.2 mM sulfide for 48 h resulted in decreases in the ATP levels in the muscle and liver. However, the energy charge in both tissues remained unchanged, and the level of lactate accumulated in the muscle was too low to have any major contribution to the energy budget of the fish. Our results reveal that B. boddaerti possesses inducible mechanisms to detoxify sulfide in an ample supply or a lack of O2. In normoxia, it detoxified sulfide to sulfate, sulfite, and thiosulfate. There were significant increases in the activities of sulfide oxidase in the muscle and liver of specimens exposed to sulfide, with that in the liver being >13-fold higher than that in the muscle. However, in hypoxia, sulfide oxidase activity in the liver was suppressed in response to environmental sulfide. In such conditions, there were significant increases in the activities of sulfane sulfur-forming enzyme(s) in the muscle and liver that were not observed in specimens exposed to sulfide in normoxia. Correspondingly, there were no changes in the levels of sulfate or sulfite in the muscle or liver. Instead, B. boddaerti detoxified sulfide mainly to sulfane sulfur in hypoxia. In conclusion, B. boddaerti was able to activate different mechanisms to detoxify sulfide, producing different types of detoxification products in normoxia and hypoxia. PMID- 15547801 TI - Age, sex, and reproductive state influence free amino acid concentrations in the fasting elephant seal. AB - Long-term fasting is a component of northern elephant seal (Mirounga angustirostris) life history requiring physiological adaptations to nitrogen conservation. Plasma free amino acids (FAAs) were determined for five elephant seal pups during the second and eighth weeks of the postweaning fast, six lactating female seals at 4-6 and 25 d postpartum, and seven sexually competitive adult male seals taken midway through the breeding season. Total FAAs declined in lactating females (11%) and pups (30%) with time fasting, but cystine concentration more than doubled in pups while decreasing by approximately 43% in lactating females. Methionine concentration significantly increased (approximately 68%) across lactation in adult females but was low for all classes of seal. Alanine was the most abundant FAA in adult males, and glycine became the dominant FAA in adult females late in lactation. Glutamine dominated the FAAs of weaned pups across the fast. Reductions in total FAAs of weanlings mirrored reductions in protein catabolism, but reductions in total FAAs also occurred in lactating females concomitant with an increase in protein catabolism. Observed variation in FAA concentrations may reflect ontogenetic requirements for certain amino acids in fasting weanlings. Similarly, increases in specific FAA concentrations across lactation may reflect variations in FAA flux resulting from the nutrient demands of lactogenesis. PMID- 15547802 TI - Respiratory chemosensitivity during wake and sleep in harbour seal pups (Phoca vitulina richardsii). AB - In this study, we examined the cardiorespiratory patterns of harbour seal pups under normoxic/normocarbic (air), hypoxic/normocarbic (15%, 12%, and 9% O2 in air), and normoxic/hypercarbic (2%, 4%, and 6% CO2 in air) conditions while awake and sleeping on land. Animals were chronically instrumented to record electroencephalogram (EEG), electromyogram (EMG), and electrocardiogram (EKG) signals, which, along with respiration (whole-body plethysmography) and oxygen consumption (VO2), were recorded from animals breathing each gas mixture for 2-4 h on separate days. Our results show that for animals breathing air, VO2 was not significantly lower during slow-wave sleep (SWS; 7.71 +/- 0.39 mL O2 min(-1) kg( 1); all measurements are mean +/- SEM) than during wakefulness (WAKE; 8.80 +/- 0.25 mL O2 min(-1) kg(-1)) and was unaffected by changes in respiratory drive. Although there was no significant fall in VO2 associated with a decrease in arousal state, breathing frequency (f(R)) did decrease (from 18.80 +/- 1.50 breaths min(-1) in WAKE to 10.40 +/- 0.49 breaths min(-1) in SWS), while the incidence of long apneas (>20 s) increased (12.76 +/- 4.06 apneas h(-1) in WAKE and 31.95 +/- 2.37 apneas h(-1) in SWS). Breathing was rarely seen during rapid eye movement (REM) sleep. Tachypnea was present at all levels of increased respiratory drive; however, hypoxia induced a dramatic bradycardia regardless of arousal state, while hypercarbia produced a tachycardia in SWS only. The hypoxic and hypercarbic chemosensitivities of harbour seal pups were similar to those of terrestrial mammals; however, unlike terrestrial mammals, where hypoxic and hypercarbic sensitivities are often reduced during SWS, the sensitivity of harbour seal pups to hypoxia and hypercarbia remained unchanged during the decrease in arousal state from WAKE to SWS. PMID- 15547805 TI - Microbiology of chronic ethmoid sinusitis: is this a bacterial disease? AB - PURPOSE: Review the microbiology of chronic rhinosinusitis and test the hypothesis that bacterial infections are not the predominant etiology for chronic rhinosinusitis. MATERIALS AND METHODS: Tissue cultures were prospectively collected from the ethmoid mucosa of 179 patients (94 had chronic rhinosinusitis and 85 had chronic rhinosinusitis with polyposis) at the time of endoscopic sinus surgery. All patients were off antibiotics for at least 1 week at the time of surgery. Both aerobic and anaerobic cultures were obtained for each specimen. RESULTS: A total of 263 isolates were identified (average: 1.5 isolates per specimen). Coagulase-negative Staphylococcus, a likely contaminant, was the most common isolate and was identified in 51% of patients. Forty percent of patients (72/179) grew pathogenic aerobic bacteria, the most common was Staphylococcus aureus (18%). Six percent of patients (10/179) grew pathogenic anaerobic bacteria. The microbiology of patients with and without nasal polyposis was not significantly different. CONCLUSION: These findings suggest that factors other than bacterial infection may play an integral role in the pathogenesis of chronic rhinosinusitis. PMID- 15547804 TI - Genetic investigation of quantitative traits related to autism: use of multivariate polygenic models with ascertainment adjustment. AB - Autism is a severe developmental disorder of unknown etiology but with evidence for genetic influences. Here, we provide evidence for a genetic basis of several quantitative traits that are related to autism. These traits, from the Broader Phenotype Autism Symptom Scale (BPASS), were measured in nuclear families, each ascertained through two probands affected by autism spectrum disorder. The BPASS traits capture the continuum of severity of impairments and may be more informative for genetic studies than are the discrete diagnoses of autism that have been used by others. Using a sample of 201 nuclear families consisting of a total of 694 individuals, we implemented multivariate polygenic models with ascertainment adjustment to estimate heritabilities and genetic and environmental correlations between these traits. Our ascertainment adjustment uses conditioning on the phenotypes of probands, requires no modeling of the ascertainment process, and is applicable to multiplex ascertainment and multivariate traits. This appears to be the first such implementation for multivariate quantitative traits. The marked difference between heritability estimates of the trait for language onset with and without an ascertainment adjustment (0.08 and 0.22, respectively) shows that conclusions are sensitive to whether or not an ascertainment adjustment is used. Among the five BPASS traits that were analyzed, the traits for social motivation and range of interest/flexibility show the highest heritability (0.19 and 0.16, respectively) and also have the highest genetic correlation (0.92). This finding suggests a shared genetic basis of these two traits and that they may be most promising for future gene mapping and for extending pedigrees by phenotyping additional relatives. PMID- 15547806 TI - Mohs micrographic surgery and the otolaryngologist. AB - OBJECTIVE: The purpose of this study was to provide an update and overview of the current indications for Mohs micrographic surgery (MMS) for cutaneous and upper aerodigestive tract malignancies. STUDY DESIGN: Literature review. METHODS: A comprehensive review of the literature from 1971 to present was completed by using the Medline data base. In addition, interviews were obtained with several otolaryngologists who had used the technique. RESULTS: MMS offers a superior alternative to standard excision in a variety of cutaneous malignancies. Overall, for the skin cancers likely to be encountered by the otolaryngologist, there is significant evidence that MMS offers increased rates of cure as well as tissue conservation. The role of MMS in upper aerodigestive tract malignancies continues to evolve and multiple issues must be addressed before its general application. CONCLUSIONS: When confronted with a difficult cutaneous malignancy, the otolaryngologist should consider MMS to optimize treatment results. PMID- 15547807 TI - Physiotherapy in patients with facial nerve paresis: description of outcomes. AB - PURPOSE: The purpose of this study was to describe changes and stabilities of long-term sequelae of facial paresis in outpatients receiving mime therapy, a form of physiotherapy. MATERIAL AND METHODS: Archived data of 155 patients with peripheral facial nerve paresis were analyzed. Main outcome measures were (1) impairments: facial symmetry in rest and during movements and synkineses; (2) disabilities: eating, drinking, and speaking; and (3) quality of life. RESULTS: Symmetry at rest improved significantly; the average severity of the asymmetry in all movements decreased. The number of synkineses increased for 3 out of 8 movements; however, the group average severities decreased for 6 movements; substantially fewer patients reported disabilities in eating, drinking, and speaking; and quality of life improved significantly. CONCLUSION: During a period of approximately 3 months, significant changes in many aspects of facial functioning were observed, the relative position of patients remaining stable over time. Observed changes occurred while the patients participated in a program for facial rehabilitation (mime therapy), replicating the randomized controlled trial-proven benefits of mime therapy in a more varied sample of outpatients. PMID- 15547808 TI - The role of viruses in idiopathic peripheral facial palsy and cellular immune response. AB - PURPOSE: The purpose of this study is to investigate the role of viruses on the idiopathic peripheral facial palsy and show the interaction of immune system. MATERIALS AND METHODS: The levels of immunoglobulin (Ig) G and IgM antibodies against to varicella-zoster virus (VZV), herpes simplex virus (HSV), cytomegalovirus (CMV), Ebstein-Barr virus (EBV), and mumps virus in venous blood taken from patients in the amount of 10 mL have been investigated by enzyme linked immunosorbent assay method. Tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), and transforming growth factor-beta (TGF-beta) were also examined. Of lymphocyte subpopulation, antibodies of CD3(+), CD4(+), CD8(+), CD19(+), and CD16(+) plus 56(+) were analyzed. RESULTS: Ten of the patients had HSV-1 IgG; 1 of the patients IgM, 5 of the patients EBV IgG, 6 of the patients VZV IgG, 1 of the patients IgM, 9 of the patients mumps IgG, 1 of the patients IgM, and finally in 7 of the patients CMV IgG antibodies were obtained. It was found that CD4(+) cell and ratio of CD4(+)/CD8(+) lower and the percentage of the CD8(+) and CD16(+) plus 56(+) cells higher compared with the control group (P < .05). The levels of TNF-alpha were lower, whereas IFN-gamma and TGF-beta1 were higher. CONCLUSION: It may be concluded from these results that VZV, HSV-1, CMV, EBV, and mumps virus play a significant role in the etiology of idiopathic peripheral facial palsy and activate the cellular immunity. PMID- 15547809 TI - Intercellular adhesion molecule-1 expression in nasal polyps treated with corticosteroid. AB - PURPOSE: The aim of this study was to compare intracellular adhesion molecule-1 (ICAM-1) expression in nasal polyp cases who were administered topical corticosteroid and in middle turbinates. MATERIALS AND METHODS: Twenty-four patients with nasal polyps were included in the study group. These patients were treated with 100 microg budesonide in each nostril twice daily for 2 months before the surgery. Twenty-one nonatopic patients with concha bullosa were included in the control group. The specimens were taken from patients undergoing endoscopic surgery. RESULTS: In polyps, significantly higher mean ICAM-1 intensity scores were found by comparison with the control turbinates. CONCLUSIONS: Corticosteroid treatment in patients with nasal polyps does not diminish ICAM-1 to that of turbinate tissue. The initiating events in the formation of nasal polyps still occur in these patients despite treatment with the topical nasal steroid. PMID- 15547810 TI - High-frequency hearing loss and middle ear involvement in rheumatoid arthritis. AB - PURPOSE: We aimed to evaluate the possible changes in the middle ears of patients with rheumatoid arthritis (RA) and to measure their hearing functions by eleven different test frequencies including standard and high frequencies. MATERIALS AND METHODS: The study group was comprised of 74 patients with RA. The control group was comprised of 45 healthy subjects. We performed speech test, tympanometry, acoustic reflex test, and standard and high-frequency pure tone audiometric tests in the study and the control groups. RESULTS: It was observed that significant increases in the thresholds of pure tones in all test frequencies for the study group versus the controls (P <.0001). In terms of the duration of the disease, hearing loss of the patients with disease duration of 1 to 5 years was higher than that of the controls and exceeded beyond the level of 20 dB especially in high frequencies starting from 10,000 Hz. In patients with disease duration of 6 to 10 years, the beginning point for hearing loss was 4,000 Hz. The patients with disease duration of 11 to 15 years and of above 16 years had high hearing thresholds in all frequencies. We found normal pressure in the middle ears of the study and the control groups. CONCLUSION: Sensorineural type hearing loss developed in patients with RA especially in high frequencies. We also determined a diminished compliance in the middle ears of those patients, but this was not in a level of a conducting type hearing loss. PMID- 15547811 TI - Anatomic findings in patients undergoing revision endoscopic sinus surgery. AB - OBJECTIVE: To report the anatomic findings that contribute to persistent sinusitis in patients requiring revision functional endoscopic sinus surgery (FESS). METHODS: Data were collected prospectively on consecutive patients requiring revision FESS at a tertiary institution over a 2-year period. Patients were evaluated with endoscopic examination of the sinonasal cavities, and computed tomography of the sinuses was performed after patients failed prolonged medical therapy for sinusitis. Information was also collected during the revision surgery. RESULTS: The most common anatomic factor associated with primary surgery failure was lateralization of the middle turbinate (78%) followed by incomplete anterior ethmoidectomy (64%), scarred frontal recess (50%), incomplete posterior ethmoidectomy (41%), and middle meatal antrostomy stenosis (39%). In addition, retained agger nasi and retained uncinate process were identified in 49% and 37% of the patients, respectively. Recurrent polyposis was seen in 37% of the patients. Other factors such as persistent sphenoid disease and sphenoid ostium stenosis were less frequent. CONCLUSION: Failure of primary FESS is most often associated with anatomic obstruction in the area of the ostiomeatal complex. Meticulous attention in this area during surgery with ventilation of obstructed anatomy as well as avoidance of scarring and turbinate destabilization may reduce the failure rate after primary FESS. PMID- 15547812 TI - Lacrimal sac rhinosporidiosis: a case report. AB - Rhinosporidiosis is a disease caused by Rhinosporidium seeberi. It usually affects the nasal mucosa and rarely the conjunctiva, lacrimal sac, tonsils, and skin. We present a case study of an isolated lacrimal sac rhinosporidiosis in an 8-year-old girl who was a migrant from Orissa, an Eastern coastal state of India. The mode of presentation and management of this case with a review of literature is discussed in brief. PMID- 15547813 TI - Facial nerve schwannomas: a report of four cases and a review of the literature. AB - OBJECTIVE: To evaluate both the surgical approaches and results of the facial nerve schwannoma cases as diagnosed in our clinic in line with the literature. MATERIAL AND METHODS: The files of 4 cases diagnosed in our clinic as facial nerve schwannoma between 1996 to 2002 were reviewed retrospectively. All the cases were evaluated with detailed history; ear, nose, and throat examination; computed tomography; and/or magnetic resonance imaging. If required, electromyography and audiometric evaluations were made. Fine-needle aspiration cytology was also performed in appropriate cases. The surgical approach used was determined depending on the tumor location and its extent. Facial nerve reconstruction was accomplished if the integrity of the nerve could not be protected. Cases were followed up for facial nerve function, complications, and recurrence. RESULTS: The tumors occurred in the vertical (n = 2), internal auditory canal (n = 1), and the peripheric segment (n = 1) of the nerve. As symptoms, facial paralysis (n = 2), hearing loss (n = 3), tinnitus (n = 1), otorrhea (n = 2), parotid mass (n = 2), and external auditory canal mass (n = 1) were observed. Radiologic investigations provided important informations. However, fine-needle aspiration cytology did not contribute to a correct preoperative diagnosis. The superficial parotidectomy approach (n = 1), retrosigmoid approach (n = 1), and transmastoid-superficial parotidectomy combined approach (n = 2) were used. The integrity of the nerve could not be protected in 3 of the cases. After the reconstruction surgery, the facial nerve function was evaluated as House-Brackmann grade 3 for all 3 cases at the mean 38.6 months follow-up time. No tumors have recurred during follow-up. CONCLUSION: Facial nerve schwannoma is a rare tumor. Through improved surgical and reconstruction techniques, postoperative morbidity is at acceptable levels. The rate of recurrence is low. It should be kept in mind for differential diagnosis of facial nerve paralysis. PMID- 15547814 TI - Primary pleomorphic liposarcoma of the parotid gland: a case report and review of the literature. AB - Although liposarcomas are the most common type of sarcoma in adults, they are rare in the head and neck region. Pleomorphic liposarcoma is the least common histologic subtype in all locations. To our knowledge, there have been only 11 reported cases of primary liposarcomas of the major salivary glands, and the pleomorphic variant arising in the parotid gland has been reported only once before. Only 2 of the 11 reported cases of liposarcoma of the major salivary glands in the literature have died of disease, and both cases were of the pleomorphic subtype. We report a case of pleomorphic liposarcoma arising in the left parotid gland of an 80-year-old woman. This case report highlights that, despite conservative surgery followed by 2 local recurrences, our patient is doing well with minimal local morbidity and no demonstrable metastases 30 months after the initial diagnosis. We also present a review of the literature on the subject. PMID- 15547815 TI - Subtotal tongue necrosis in delayed diagnosed giant-cell arteritis: a case report. AB - Giant-cell arteritis (GCA) is a chronic systemic vasculitis of large- and medium sized vessels, mainly affecting elderly patients. Headache, vision impairment, jaw claudication, and scalp tenderness are common symptoms. However, diagnosis can be difficult because GCA can affect almost every vascular pathway and lead to a variety of possible manifestations. We report the case of a belated diagnosed GCA, resulting in nearly complete necrosis of the mobile part of the tongue, visual impairment, and neurologic as well as intestinal ischemic symptoms. Aggressive immunosuppressive treatment resolved the symptoms, but the patient remained severely morbid because of bilateral necrosis of the mobile part of the tongue. In any case of unclear ischemic symptoms in an elderly patient, one must keep GCA in mind as the possible culprit disease. PMID- 15547816 TI - Rapid evolution of acute mastoiditis: three case reports of otogenic meningitis in adults. AB - Otogenic meningitis is the most common intracranial complication of neglected otitis media. In the western world, such complications seldom occur in children and young adults and are extremely rare in adults and elderly people. The current use of antibiotics and of more sophisticated surgery has greatly diminished the incidence of otogenic meningitis in comparison with the past. This has resulted in physicians having less experience concerning diagnosis and treatment of this complication. The authors reported 3 consecutive cases of otogenic meningitis in adults, which occurred in the space of 3 months after a 6-year absence of such pathology at their institution. In all 3 cases, conventional antibiotic therapy proved ineffective; the course of the disease worsened rapidly in contrast with the lack of symptoms during the period before treatment. Emergency surgical treatment was mandatory. PMID- 15547818 TI - Postpericardiotomy syndrome after minimally invasive repair of pectus excavatum. AB - Minimally invasive repair of pectus excavatum (MIRPE) was first reported in 1998 and has gained wide acceptance since then. A 17-year-old girl who had undergone thoracotomy and cardiac surgery for transposition of great vessels at the age of 18 months presented with a deep, long pectus excavatum with asymmetry. After initial uneventful postoperative clinical course after MIRPE, the patient had bilateral pleural and pericardial effusion on the sixth postoperative day. Suspecting postpericardiotomy syndrome, systemic steroids were administered, and the symptoms resolved without affecting wound healing. Manifestation of a pericardial effusion combined with bilateral pleural effusion after MIRPE, especially in patients after cardiac surgery, may indicate a postpericardiotomy syndrome that can be treated successfully by intravenous steroids. PMID- 15547819 TI - Ruptured hemorrhagic cyst in an undescended ovary. AB - The authors report on a patient with an uncommon congenital anomaly of bilateral undescended ovaries and tubes. She presented with an acute abdomen at 13 years of age. A computed tomography scan showed a cystic mass suspicious for congenital intestinal duplication. At the time of laparoscopy, she was found to have a bicornuate uterus and bilateral undescended ovaries with a ruptured right hemorrhagic ovarian cyst. This case shows that although a ruptured ovarian cyst can occur in such malpositioned organs, it can pose difficult diagnostic challenges. Suppression of the ovaries with the combined oral contraceptive pill may help to prevent further ovarian cyst development. PMID- 15547820 TI - Sustained inquiry: in the clinic and at the bench. PMID- 15547821 TI - Thymic transplantation for complete DiGeorge syndrome: medical and surgical considerations. AB - BACKGROUND/PURPOSE: Complete DiGeorge syndrome results in the absence of functional T cells. Our program supports the transplantation of allogeneic thymic tissue in infants with DiGeorge syndrome to reconstitute immune function. This study reviews the multidisciplinary care of these complex infants. METHODS: From 1991 to 2001, the authors evaluated 16 infants with complete DiGeorge syndrome. All infants received multidisciplinary medical and surgical support. Clinical records for the group were reviewed. RESULTS: Four infants died without receiving a thymic transplantation, and 12 children survived to transplantation. The mean age at time of transplantation was 2.7 months (range, 1.1 to 4.4 months). All 16 infants had significant comorbidity including congenital heart disease (16 of 16), hypocalcemia (14 of 16), gastroesophageal reflux disease or aspiration (13 of 16), CHARGE complex (4 of 16), and other organ involvement (14 of 16). Nontransplant surgical procedures included central line placement (15 of 16), fundoplication or gastrostomy (10 of 16), cardiac repair (10 of 16), bronchoscopy or tracheostomy (6 of 16), and other procedures (12 of 16). Complications were substantial, and 5 of the 12 transplanted infants died of nontransplant-related conditions. All surviving infants have immune reconstitution, with follow-up from 2 to 10 years. CONCLUSIONS: Although the transplantation of thymic tissue can restore immune function in infants with complete DiGeorge syndrome, these children have substantial comorbidity. Care of these children requires coordinated multidisciplinary support. PMID- 15547822 TI - Expression of apoptosis-related genes after fetal tracheal occlusion in rabbits. AB - BACKGROUND/PURPOSE: Late-gestation lung remodeling is associated with alveolar type II cell apoptosis early in the saccular stage (day 28 in fetal rabbits). Intrauterine tracheal occlusion (TO), a potent stimulus of fetal lung growth and maturation, significantly increases type II cell apoptosis. The aim of this study was to determine the effect of fetal TO on the spatiotemporal expression of key apoptosis-related signaling molecules. METHODS: Tracheal occlusion of fetal rabbits was performed at gestational day 25 (term, 31 days), and apoptotic gene expression was studied between days 26 and 28. RESULTS: At days 26 and 27, the protein levels of Fas and Fas-ligand (FasL) in lung lysates were similar in TO fetuses and sham-operated controls. At day 28, however, synchronous with the onset of TO-induced pulmonary distension and type II cell apoptosis, the FasL protein content was 8-fold higher in TO lungs compared with controls (P < .01), whereas Fas levels were comparable. In contrast, Bax and Bcl-2 protein levels were similar in TO and control fetuses at all time-points. TO significantly increased the cellular concentration of immunoreactive FasL in type II cells and bronchial epithelial Clara cells. Furthermore, bronchoalveolar lavage fluid (BAL) from TO fetuses at day 28 induced significantly more type II cell apoptosis in vitro compared with control BAL, an effect that was inhibited by neutralizing anti-FasL antibody. CONCLUSIONS: Our findings show that TO results in time specific increase of both cellular and soluble FasL in fetal lungs and implicate the Fas/FasL pathway as a pivotal autocrine and/or paracrine regulator of TO- induced type II cell apoptosis. PMID- 15547823 TI - Systemic hypertension associated with venovenous extracorporeal membrane oxygenation for pediatric respiratory failure. AB - BACKGROUND/PURPOSE: Arterial hypertension (HTN) is common in neonates on venoarterial (VA) extracorporeal membrane oxygenation (ECMO), but HTN in pediatric venovenous (VV) ECMO has not been well described. The authors noted HTN in their VV ECMO experience and hypothesized that HTN was associated with fluid status, steroid use, and renal insufficiency. METHODS: Records of 50 patients receiving VV ECMO for respiratory failure were reviewed. HTN was defined as systolic blood pressure greater than 95th percentile for age for > or =1 hour, unresponsive to sedation/analgesia. Hypertensive index (HI) is defined as total hypertensive hours per total ECMO hours. Fluid status was estimated by a fluid index (FI = total fluid balance during ECMO per ECMO hours per weight). RESULTS: Forty-seven of 50 patients (94%) had HTN. Median HI was 0.21 (range, 0.01 to 1.0). Thirteen patients had renal insufficiency, 39 received steroids, and 23 received continuous venovenous hemofiltration (CVVH). There was no association between HI and FI, steroid use, or renal insufficiency. Thirty-three patients were treated for HTN, often requiring multiple agents. Bleeding complicated the course of 18 patients, and HI was significantly higher in those patients (P = .03). HI was not different between survivors (37 of 39 with HTN) and nonsurvivors (10 of 11 with HTN). CONCLUSIONS: Hypertension is a common complication associated with VV ECMO with unclear etiology. HTN was frequently difficult to control. This study emphasizes the need for the development of treatment protocols to decrease the incidence, severity, and associated morbidity. Improved insight into the etiology of HTN associated with pediatric VV ECMO, including evaluation of the renin-angiotensin system, would help guide therapy. PMID- 15547824 TI - Surfactant replacement therapy on ECMO does not improve outcome in neonates with congenital diaphragmatic hernia. AB - BACKGROUND PURPOSE: Respiratory failure in neonates with congenital diaphragmatic hernia (CDH) may in part be caused by a primary or secondary surfactant deficiency. Knowledge of the optimal approach to surfactant replacement in neonates with CDH and respiratory failure is limited. The aim of this study was to determine if surfactant replacement on extracorporeal membrane oxygenation (ECMO) results in improved outcomes in neonates > or =35 weeks' gestation with unrepaired CDH. METHODS: Using the CDH Study Group Registry, the authors identified 448 neonates with CDH who were > or =35 weeks' gestation, had no major anomalies, were treated with ECMO within the first 7 days of life, and underwent repair on or after ECMO therapy. Patients in 2 groups were compared: group 1 (- Surf, n = 334) consisted of patients who received no surfactant and group 2 (+ Surf, n = 114) consisted of patients who received at least 1 dose of surfactant while on ECMO. An analysis of all patients in both groups was performed. Additionally, subgroup analyses stratified by gestational age were performed for patients 351/7 to 366/7 weeks' gestation and for patients > or =37 weeks' gestation. Primary end-points for the study were survival and length of ECMO run. Secondary end-points were length of intubation, need for supplemental oxygen at 30 days of life, and at discharge to home. Demographic, clinical, and outcome variables were examined using Fisher's Exact tests for categorical variables and using unpaired t tests for continuous variables. Odds ratios were calculated for categorical end-point variables. RESULTS: Demographic and clinical variables were similar between groups. Analyses of aggregate data showed no significant differences between groups in length of ECMO run, survival, number of days intubated, and percent of patients requiring supplemental oxygen at 30 days or discharge. Subgroup stratification by gestational age did not show significant differences between groups in any of the outcome variables. CONCLUSIONS: The data from this study suggest that surfactant replacement on ECMO for neonates with congenital diaphragmatic hernia does not provide significant benefit in the infant's clinical course with respect to survival, length of ECMO course, length of intubation, or subsequent need for supplemental oxygen. PMID- 15547825 TI - Drainage, fibrinolytics, or surgery: a comparison of treatment options in pediatric empyema. AB - BACKGROUND: The current treatments of pediatric empyemas include tube thoracostomy with or without the instillation of fibrinolytics, video-assisted thoracoscopic surgery (VATS), and open thoracotomy with decortication. Whereas success has been reported for all of these techniques, VATS has been suggested as the best method because of decreased length of stay. METHODS: A chart review of children who presented with parapneumonic effusions from February 2000 to June 2002 was conducted. The patients were divided into 4 groups depending on the treatment received: group I, chest tube alone (n = 18); group II, chest tube and fibrinolytics (n = 24); group III, chest tube, fibrinolytic, and surgery (n = 5); and group IV, surgery alone (n = 6). Preadmission, in-hospital, and outcome variables for the groups were recorded and compared using the Kruskall-Wallis test, with a P value less than .05 considered significant. All the patients who received fibrinolytics (group II and III) were grouped into subjects who received immediate transpleural fibrinolytics versus those who received fibrinolytics 48 hours after chest tube insertion. Length of stay (LOS), need for surgery, and hospital costs were compared between the early and late fibrinolytic groups using the Wilcoxon rank-sum test, with a P value less than .05 considered significant. RESULTS: Comparison of duration of symptoms, duration of preadmit antibiotics, initial white blood cell count, total lymphocyte count, and antibiotics showed no significance among the 4 groups. When comparing outcome variables, the "nonsurgery groups" (groups I and II) had shorter LOS, intensive care unit stay, and hospital charges when compared with the "surgery groups" (groups III and IV). The timing of fibrinolytic instillation (immediate versus later) did not significantly affect in the LOS, hospital charges, or the tendency to need surgery eventually in the patients who received intrapleural fibrinolytics (group II and III combined). LOS was predicted by preadmit duration of symptoms (P = .025) and overall duration of fever (P < .01). The level of pleural glucose seemed to be predictive of need for surgery (P = .015). Overall, 11 of 54 children (20.2%) eventually needed surgery. CONCLUSIONS: Tube drainage with intrapleural instillation of fibrinolytics can be performed successfully in a large number of children with empyemas. Ultrasound characterization of the fluid and, perhaps, glucose levels may guide surgical versus nonsurgical therapy. In centers in which percutaneous drainage and tissue plasminogen activator are available, this option may be a safe and less costly alternative to surgery. PMID- 15547826 TI - Is routine dilatation after repair of esophageal atresia with distal fistula better than dilatation when symptoms arise? Comparison of results of two European pediatric surgical centers. AB - BACKGROUND/PURPOSE: The aim of this study was to determine whether routine dilatation of the anastomosis after repair of an esophageal atresia with distal fistula (EADF) is superior to a wait-and-see policy with dilatation only when symptoms arise. METHODS: The records of 100 consecutive patients operated on for EADF in 2 European pediatric surgical centers (A [n = 63], B [n = 37]) were reviewed. In center A, dilatation of the anastomosis was carried out in symptomatic cases only, whereas in center B dilatation was begun 3 weeks postoperatively and repeated every 1-3 weeks until a stable diameter of 10 mm was reached. Particular attention was paid to the number of dilatations per patient, dilatation-related complications, and differences in results after 2 years. RESULTS: The patient materials of both centers did not differ with respect to the incidence of prematurity, tracheomalacia, gastroesophageal reflux (GER), and major postoperative complications. The incidence of associated anomalies was higher in center B (P < .05). In center A, 26 of 63 patients underwent dilatation; in center B, all 37 patients were dilated (P < .05). Median number of dilatations per patient was 4 in center A and 7 in center B (P < .05). In center A, 23 of 26 and in center B, 20 of 37 of the patients received medical treatment for GER at the time of the dilatations. Dilatation-related complications developed in 7 of 26 patients of center A and in 3 of 37 patients in the center B (P value, not significant). The median primary hospital stay was 24 days in center A and 33 days in center B (P < .05), median secondary hospital stay for dilatation was 6 days in center A and 13 days in center B (P < .05). After 2 years of follow-up, the incidence of dysphagia, respiratory problems, or bolus obstruction did not differ significantly between the 2 centers. CONCLUSIONS: A wait-and-see policy and dilatations based on clinical indications for patients with repaired EADF is superior to routine dilatations. It appears that more than half of the patients do not require dilatations at all. PMID- 15547827 TI - Image-guided percutaneous gastrostomy in neonates with esophageal atresia. AB - BACKGROUND: Neonates with esophageal atresia may require a gastrostomy before definitive repair. Most surgeons do this procedure using the Stamm technique through a laparotomy. The authors describe a new technique for percutaneous placement of a gastrostomy in these infants and report their preliminary results. METHODS: For children with esophageal atresia and a tracheoesophageal fistula (TEF), the air-filled stomach is localized fluoroscopically and accessed using a standard percutaneous technique. For those with pure esophageal atresia, a transhepatic needle is used to instil air into the stomach. Once the stomach is distended, the gastrostomy tube is inserted under fluoroscopy. RESULTS: Fourteen neonates with esophageal atresia had a percutaneous gastrostomy tube placed. Eleven had esophageal atresia and a TEF, and 3 had pure esophageal atresia with a gasless abdomen requiring the transhepatic approach. There were no intraoperative or major postoperative complications, but there were 5 minor early postoperative complications and 2 minor late postoperative complications. CONCLUSIONS: Percutaneous gastrostomy insertion is a safe technique for neonates with esophageal atresia and can be used even in children with pure esophageal atresia who have a gasless abdomen. This technique does not require laparotomy and appears to be associated with a low rate of complications. PMID- 15547828 TI - Gastric outlet obstruction secondary to acid ingestion in children. AB - BACKGROUND/PURPOSE: Gastric outlet obstruction (GOO) is a well-known complication of acid ingestion. However, most reports deal with adults. In this report, the authors present their experience with the treatment of acid-induced GOO in children. METHODS: The records of patients admitted for unintentional ingestion of corrosive agents between 1980 and 2002 were reviewed retrospectively. Data concerning age at ingestion, type of ingested substance, time between ingestion and the first signs of GOO, weight loss, treatment, complications, duration of hospital stay, and long-term follow-up were reviewed. RESULTS: GOO was not observed in any of the children admitted for alkaline ingestion, whereas GOO developed in 8 of 98 children (8.2%) in a mean period of 26.7 +/- 10 days after the ingestion of acid substances. Presenting symptoms were frequent nonbilious vomiting and marked weight loss. All had pyloric obstructions in the upper gastrointestinal series and required surgical intervention. Gastrojejunostomy was the operation of choice for all patients. Oral feedings were started on the third postoperative day. The complications were wound infection in 1 and upper gastrointestinal bleeding in another in the early postoperative period. Mean follow-up is 8.33 +/- 4.45 (4.8-18.7) years. No late complications such as marginal ulcus or stricture at the anastomosis site were observed in the series. CONCLUSIONS: Treatment of GOO with gastrojejunostomy gives good long-term results in children. This procedure is safe and causes minimal morbidity particularly in patients without extensive gastric damage. PMID- 15547829 TI - Annular pancreas in children: a recent decade's experience. AB - PURPOSE: Annular pancreas is a recognized cause of duodenal obstruction in children. The authors sought to characterize the clinical, radiologic, and prognostic findings in this disorder through a 10-year review of all patients with annular pancreas. METHODS: A retrospective review of all annular pancreas cases identified between 1993 and 2002 at 2 tertiary pediatric surgical centers was completed. Factors analyzed included patient demographics, presenting symptoms, associated anomalies, radiologic findings, operation performed, postoperative outcomes, and complications. RESULTS: Sixteen patients were identified. Four patients (25%) were premature. Twelve patients (75%) presented during the first week of life and the remainder within the first year. All patients presented with vomiting, which was nonbilious in 15 (94%). Five patients (31 %) had chromosomal anomalies, and 6 (38%) had other major congenital malformations. Eleven patients (69%) presented with partial duodenal obstruction. Operations performed were duodenoduodenostomy in 14 (88%), duodenojejunostomy in 1 (6%), and Ladd's procedure in 1 (6%). Enteral feedings were started, and full enteral feeding was achieved at an average of 8.4 days and 15.9 days after operation, respectively. All patients survived and were discharged after an average hospitalization of 24.0 days. CONCLUSIONS: Patients with annular pancreas have preampullary duodenal obstruction, which is more commonly partial. Duodenoduodenostomy is the appropriate treatment. Prognosis is excellent, despite the frequent association with chromosomal anomalies and major congenital malformations. PMID- 15547830 TI - The aberrant expression of HLA-DR in intrahepatic bile ducts in patients with biliary atresia: an immunohistochemistry and immune electron microscopy study. AB - BACKGROUND/PURPOSE: Although the precise pathogenesis for the destruction of the intrahepatic biliary tract in a patient with biliary atresia (BA) is still unclear, it is known that progressive destruction of the intrahepatic biliary tract is responsible for poor long-term outcome after surgical treatment. In this study, the authors evaluated the expression of HLA-DR in liver tissue of patients with BA to explore its role in the pathogenesis of this disease. METHODS: Liver biopsy specimens from patients with BA (n = 14) and normal liver biopsies (n = 12) were studied immunohistochemically and were assessed immune electron microscopically to detect the expression of HLA-DR. All patients underwent follow up for at least 3 months after surgery evaluating the short-term postoperative outcome by monitoring the serum bilirubin level during jaundice persistency. RESULTS: The immunohistochemical examinations showed in 10 of 14 liver specimens in the BA group a positive expression of HLA-DR. This expression was mainly detected in bile ducts of the central portal area rather than in proliferated ones in more peripheral areas. None of the specimens from the control group showed positive expression. Eight of 14 specimens in the BA group showed immunoelectron microscopic depositions of colloidal gold in dull and swollen ductules microvilli. This was not observed in the control group. In patients with BA there was an inverse relationship between short-term postoperative outcome and expression of HLA-DR in intrahepatic ductules. CONCLUSIONS: Our findings show that there is an aberrant expression of HLA-DR in microvilli of the ductules in patients with BA, and this abnormal expression is inversely related to the short term postoperative outcome. It is suggested that aberrant HLA-DR expression in bile ductules may play a pathogenic role in progressive damage to the intrahepatic bile ducts in BA. PMID- 15547832 TI - Renal function and outcome following salvage surgery for bilateral Wilms' tumor. AB - PURPOSE: Maintaining adequate renal function without compromising cure in children with bilateral Wilms' tumor is surgically demanding and challenging. The aim of this study was to assess renal function and outcome in children treated in one institution for bilateral Wilms' disease by renal salvage procedures. METHODS: This study is a retrospective review of all patients with histologically proven bilateral nephroblastoma who underwent a renal salvage procedure between November 1973 and June 2002. The median follow-up time was 52 months (range, 8 to 326 months). RESULTS: Twenty-three patients were included in the study. The median age at diagnosis was 19 months (range, 5 to 65 months). Patients who presented before 1982 (n = 5) were treated surgically first followed by chemotherapy. The remainder (n = 18) received initial chemotherapy before the operation. Of the 46 kidneys, 18 had a nephrectomy. The remaining 28 underwent a renal salvage procedure. At follow-up, 19 patients had good renal function, 2 had satisfactory function, and 2 had renal failure. Seventeen of the 23 children are alive and tumor free at follow-up (74%). Four children died of distant metastases and 2 of renal failure. Local recurrence in the salvaged kidney was detected in 1 patient 12 years after surgery after the commencement of immunosuppression for renal transplantation. CONCLUSIONS: These results confirm that renal salvage procedures, in combination with chemotherapy, are a safe and effective way of treating children with bilateral Wilms' disease. In addition, renal salvage surgery can maintain satisfactory renal function in the majority of these patients without an increased risk of local recurrence. When transplantation is required, the remaining native kidney should be removed to prevent tumor recurrence. PMID- 15547831 TI - Laparoscopic-assisted total cyst excision of choledochal cyst and Roux-en-Y hepatoenterostomy. AB - PURPOSE: The aim of this study was to describe the technical experience and outcome in laparoscopic-assisted total cyst excision of choledochal cyst with Roux-en-Y hepatoenterostomy. METHODS: Thirty-five patients with choledochal cyst were studied. Their age ranged from 3 months to 9 years (average age, 3.6 years). The choledochal cysts were cyst type in 33 cases and fusiform type in the other 2 cases. Four trocars were utilized with 3- to 5-mm instrumentation. Under laparoscopic guidance, the gallbladder and the dilated bile duct were excised completely. The Roux-en-Y jejunojejunostomy was performed extracorporeally by exteriorizing the jejunum through the extending umbilical incision (1.5 to 2 cm), and an end-to-side hepaticojejunostomy was carried out intracorporeally by the hand suture methods. RESULTS: Average duration of operation was 4.3 hours (range, 3.5 to 7.6 hours), intraoperative blood loss was 5 to 10 mL, and 8 of the 35 patients had associated hepatic ductal stenosis and underwent laparoscopic excision of the cyst and ductoplasty. In 1 of the 8 cases, bile leak was noticed from day 1 through 26 postoperatively. The postoperative course was uneventful in the other 34 patients with a hospital postoperative stay ranging from 3 to 6 days. There were no postoperative complications in the 3-month to 1(1/2)-year follow-up. CONCLUSIONS: Laparoscopic-assisted total cyst excision with Roux-en-Y hepatoenterostomy is feasible for the treatment of choledochal cyst in children. PMID- 15547833 TI - Different pattern of matrix metalloproteinases expression in alveolar versus embryonal rhabdomyosarcoma. AB - BACKGROUND/PURPOSE: The matrix metalloproteinases (MMPs), a family of enzymes that degrade the extracellular matrix (ECM), are important in neoplastic cell invasion and metastasis. Data for rhabdomyosarcoma (RMS), the most frequent soft tissue sarcoma of childhood, are lacking. The aim of this study was to assess their expression in this tumor and to evaluate the correlation with clinicopathologic parameters. METHODS: Immunohistochemical expression of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, TIMP-1, and TIMP-2 was investigated in 33 human RMSs, 12 alveolar, and 21 embryonal histologic subtypes (12 high risk and 9 low/standard risk). Evaluation of the results was based on the percent of positive neoplastic cells and on staining intensity (negative, moderate, and strong). In situ zymography was carried out on 4 frozen RMS samples (2 alveolar and 2 high-risk embryonal). RESULTS: Alveolar type showed a stronger MMP-1, -2 and -9 expression in comparison with embryonal (P = .006, P <.001, and P <.001, respectively). Intratumoral vessels and perivascular ECM were positive for MMP-9 in the majority of RMSs. Both TIMPs had negative results. CONCLUSIONS: Gelatinases MMP-2 and MMP-9 and collagenase MMP-1 overexpression seem to contribute to the more aggressive phenotype of alveolar rhabdomyoblastic cells. Further characterization of the expression of MMPs and consequent utilization of their inhibitors in aggressive alveolar RMSs might lead to the development of novel anticancer therapies. PMID- 15547834 TI - Laparoscopic versus open appendectomy in children with uncomplicated and complicated appendicitis. AB - PURPOSE: To address whether laparoscopic appendectomy could be an alternative to conventional open appendectomy in children with complicated appendicitis as well as uncomplicated appendicitis, a retrospective study comparing laparoscopic and open appendectomies was performed. METHODS: One hundred patients who were treated by immediate appendectomy, either laparoscopically or by the open method, between May 2000 and August 2003 were included in the study. There were 53 patients in the laparoscopic appendectomy group and 47 patients in the open appendectomy group. RESULTS: The operating time was significantly longer for laparoscopic appendectomy than for open appendectomy (P < .001). The length of hospital stay was significantly shorter in laparoscopic appendectomy in patients with uncomplicated appendicitis (P = .001). Thirteen of the 100 patients (13.0%) had 15 postoperative complications including wound infection (n = 8), intraabdominal abscess (n = 4), stitch abscess (n = 2), and small bowel obstruction (n = 1). In both uncomplicated and complicated appendicitis, there was no significant difference between laparoscopic and open appendectomies in the complication rates, and the incidences of each complication did not differ between the procedures. Among the 14 patients with generalized peritonitis, postoperative complications were seen in 5 patients (35.7%). Although the presence of generalized peritonitis was associated significantly with postoperative complications (P = .017), there was no significant association between the procedure and complications. Overall treatment costs were increased by 26.0% in laparoscopic appendectomy. CONCLUSIONS: Laparoscopic appendectomy should remain an option in children with uncomplicated and complicated appendicitis, and when laparoscopy is selected, consideration of the advantages and disadvantages of the procedure is essential. PMID- 15547835 TI - Experience of long-term synbiotic therapy in seven short bowel patients with refractory enterocolitis. AB - BACKGROUND/PURPOSE: Probiotic and prebiotic therapies are potent new strategies to treat various intestinal diseases, including inflammatory bowel disease and viral and bacterial infections. Synbiotics is defined as the combined use of probiotics and prebiotics and is expected to have a stronger effect on intestinal diseases than probiotics or prebiotics alone, but there has been no report of its clinical application. The authors designed a protocol for synbiotic therapy composed of Bifidobacterium breve, Lactobacillus casei, and galactooligosaccharides and preliminarily ascertained its clinical effects in humans. METHODS: This protocol of synbiotic therapy was applied for more than 1 year to 7 malnourished patients with short bowels who suffered from refractory enterocolitis. RESULTS: The therapeutic protocol improved the intestinal bacterial flora (inducing the domination by anaerobic bacteria and suppressing the residence of pathogenic bacteria) and increased short chain fatty acids in the feces (from 27.8 to 65.09 micromol/g wet feces). All patients but 1 accelerated their body weight gain, and 5 patients showed increased serum rapid turnover proteins. CONCLUSIONS: This protocol for synbiotic therapy might be a potent modulator of intestinal flora and a promising strategy to treat short bowel patients with refractory enterocolitis. PMID- 15547836 TI - Trefoil factor family peptide 3 prevents the development and promotes healing of ischemia-reperfusion injury in weanling rats. AB - BACKGROUND/PURPOSE: Although the pathogenesis of necrotizing enterocolitis (NEC) is not completely defined, ischemia appears to be one of the most important causative factors. Trefoil factor family peptide 3 (TFF3) is a peptide normally expressed in the small bowel and colon and is involved in the maintenance and repair of mucosal integrity. The authors hypothesized that monomeric (TFF3 Ser57) and dimeric (TFF3 Cys57) recombinant TFF3 may prevent the development and accelerate healing of intestinal ischemia-reperfusion injury in weanling rats. METHODS: Intestinal injury was induced in 18-day-old rats by occlusion of the superior mesenteric vessels for 60 minutes. To examine the protective effect, rats were given 3 microg/g of TFF3 Ser57 or TFF3 Cys57 by subcutaneous or enteral administration 30 minutes before the vascular occlusion. To examine the healing effect, rats were given 3 microg/g of TFF3 Ser57 or TFF3 Cys57 by subcutaneous or enteral administration 60 minutes after the beginning of reperfusion. Samples from small bowel and colon were collected for morphometric analysis after 3 hours of reperfusion. Mucosal damage was assessed by the Chiu score. RESULTS: Both forms of TFF3 reduced the amount of damage when administered before the ischemia. Administration of TFF3 Ser57 and TFF3 Cys57 after the beginning of reperfusion significantly increased the villous height and decreased the Chiu score in the small intestine and colon. CONCLUSIONS: TFF3 Ser57 monomer and TFF3 Cys57 dimer prevent the development and promote healing of ischemia-reperfusion injury in weanling rats. There are no differences between the routes of administration of TFF3. PMID- 15547837 TI - Placental vascular compromise in jejunoileal atresia. AB - BACKGROUND/PURPOSE: The mechanisms of intrauterine vascular disruptions that result in the development of jejunoileal atresia (JIA) are not fully understood. Monochorionic twinning with fetal death of a cotwin is known to be correlated with the development of JIA in the survivor through placental communication. The aim of this study was to evaluate whether other placental vascular compromises might contribute to the development of JIA. METHODS: Forty-five newborns (23 boys and 22 girls) who were treated for JIA at Tsukuba University Hospital from 1978 to 2003 were reviewed. Placental findings were informative in 23 cases. RESULTS: No or slight abnormality of the placenta was found in 19 cases. Significant placental abnormalities were found in 4 patients who also had a low birth weight. One patient with apple peel atresia (APA) had excessive torsion of the umbilical cord (UC), which was inserted at the margin of the placenta, and there was an adjacent area of infarction. One patent with multiple atresia (MA) was a surviving monochorionic twin with intrauterine fetal death of the other. Another case of MA showed marginal insertion of the UC. Severe placental abnormalities including wide infarction, cyst formation, and marginal insertion of the UC were found in 1 case of MA. These 3 cases of MA were complicated with other anomalies including brain anomaly. CONCLUSIONS: Placental vascular compromises were involved infrequently in JIA but might possibly be responsible for the development of JIA as well as associated anomalies and a low birth weight as chronic insults since an early stage of gestation in some cases. PMID- 15547838 TI - Associated congenital anomalies in patients with anorectal malformations--a need for developing a uniform practical approach. AB - BACKGROUND/PURPOSE: The aim of this study was to compare the incidences of various associated anomalies among subjects with high and low anorectal malformation (ARM), study the coexistence of these anomalies, and compare their frequency with those quoted in the existing literature. METHODS: Ten years of data (from January 1993 through December 2002) on the 416 subjects from our center with anorectal malformations (ARM) were evaluated. The subjects with supra or translevator rectal pouch (radiologic or operative findings) were classified as high ARM and the rest as low ARM. The associated anomalies studied were vertebral, skeletal, renal, cardiac, vesicoureteric reflux (VUR), gastrointestinal, tracheoesophageal fistula, spinal, genital, and miscellaneous. For a detailed comparison, 4 subgroups were made: boys with high ARM, boys with low ARM, girls with high ARM, and girls with low ARM. Statistical methods were used for intergroup comparison. RESULTS: There were 68% male and 32% female subjects. Overall, 58% of subjects had high ARM (supralevator and translevator). Our attempt to compare the incidences of anomalies in our study population with those in other populations was defeated because of nonuniformity of classification and investigation of these anomalies in different studies. We found a frequent clubbing of anomalies affecting distantly developing anatomic regions in patients with ARM in many studies. The highest incidence of associated anomalies was found among boys with high ARM. The male subjects also had significantly more genital anomalies in association with low ARM and gastrointestinal tract anomalies in association with high ARM. On the other hand, girls had more urologic anomalies with high ARM and VUR with low ARM. CONCLUSIONS: Except for a low incidence of spinal anomalies (8%) and of VUR (1.7%), the incidences of most associated anomalies in our study were comparable with those of the earlier studies. Additionally, the girls with high ARM were observed to have significantly more urologic anomalies compared with the boys with high ARM in contrast to the results of the earlier reports. The existence of anomalies in distantly developing anatomic regions in patients with ARM supports the possibility of a "generalized" insult during embryogenesis rather than a ("localized") defect. It was observed that the boys with ARM are more likely to suffer morbidity because of frequent occurrence of multiple associated anomalies, and it may be worthwhile to evaluate of the role of sex chromosome in relation to ARM. The authors are also of the view that there is a need for more uniformity in classification of the anomalies and in their diagnostic approach because various reported studies have differed so widely on these aspects that any interstudy comparison is difficult or not feasible. PMID- 15547839 TI - Dynamic esophageal lengthening for long gap esophageal atresia: experience with two cases. AB - This report describes two infants with long gap esophageal atresia managed by dynamic esophageal lengthening. Preliminary traction on the atretic ends of the esophagus permitted subsequent primary anastomosis under significant tension. One patient has done well, while the other developed an anastomotic stricture requiring resection. The esophagus was preserved in both infants. PMID- 15547840 TI - Vocal fold paralysis and progressive cricopharyngeal stenosis reversed by cricopharyngeal myotomy. AB - Three infants with progressive upper esophageal stenosis had bilateral vocal fold paralysis. The patients were apparently normal at birth and without neurologic abnormality. Cricopharyngeal myotomy, followed by serial dilatations, relieved esophageal stenosis and restored the swallowing function. Vocal fold mobility, to the authors' surprise, also recovered after myotomy in 2 patients. Vocal paralysis persisted in the first patient encountered, although this case was complicated by longstanding tracheostomy. Decannulation in this case was successful at 4 years only after arytenopexy and cricoid interposition grafting. Direct laryngoscopy findings showed weak abductive motion of 1 vocal fold at 14 year follow-up. Loss of vocal function secondary to constriction by a cricopharyngeal band has been previously undocumented. The potential reversibility of both vocal paralysis and esophageal stenosis by surgical cricopharyngeal myotomy is of significant clinical importance. PMID- 15547841 TI - Pulmonary torsion after cardiac surgery in two infants: review of pediatric literature. AB - Torsion of a lung or a lobe (LT) is a severe, sometimes life-threatening event that may occur spontaneously, after trauma, or after cardiac or thoracic surgery. The authors report on 2 prematurely born neonates who had LT after cardiac surgery. Both patients successfully underwent pulmonary lobectomy, which seems to be the best surgical approach. Given that careful anatomic unfolding of the lung and its reinflation under vision at the end of a cardiac or thoracic operation is deemed crucial to avoid LT, the authors suggest that, in case of a complete pulmonary fissure and/or free long bronchovascular pedicle, lobe fixation should be accomplished, too. Because of its rarity, we could find only 6 well-documented reports of LT diagnosed in children, whereas another 3 cases were quoted without clinical details. The pediatric literature is reviewed. PMID- 15547842 TI - A very rare consequence of steroid therapy: ileal perforation in a patient with familial mediterranean fever. AB - Intestinal perforation rarely occurs in children with familial Mediterranean fever (FMF). When this does happen, it is the result of untreated intestinal obstruction caused by compression from peritoneal adhesions. Intestinal perforation is a well-known complication of steroid therapy in all ages. The duodenum is affected most frequently, but perforation may also occur in other parts of the small intestine and, very rarely, the colon. Intestinal wall changes that occur in chronic FMF may promote the harmful effects of steroids. Here we present an unexpected complication, ileal perforation, in an 8-year-old boy who was taking prednisolone for FMF-related arthritis. PMID- 15547844 TI - Calcifying fibrous pseudotumor of lung: a previously unreported entity. AB - Calcifying fibrous pseudotumor (CFP) is one of the most unusual benign tumors of childhood and is located mostly in soft tissues, pleura, and peritoneum. The first case of pulmonary CFP in a 7-year-old boy is presented. The clinicopathologic features of this rare entity are discussed with special emphasis on histopathologic features and prognosis including a brief literature review. There are no presumptive clinical and laboratory findings, including tumor markers and imaging techniques, that distinguish CFP from other space occupying lung lesions. The most important diagnostic aid is to bear this entity in mind when a child presents with an unexplained pulmonary mass. Total excision is mandatory for the appropriate diagnosis and treatment. PMID- 15547843 TI - Dieulafoy's lesion and segmental dilatation of the small bowel: an uncommon cause of gastrointestinal bleeding. AB - Segmental dilatation of the bowel and Dieulafoy's lesions are uncommon but important causes of gastrointestinal bleeding. These conditions are well described in adults but can occur in children as well. Here, the authors present a case of a boy with gastrointestinal bleeding from an ulcerated Dieulafoy's lesion found concomitantly within a segment of idiopathically dilated ileum. The simultaneous occurrence of both of these lesions in a symptomatic child has not been reported previously. PMID- 15547845 TI - Ascended testis in three brothers with Pelizaeus Merzbacher syndrome. AB - Ascended testis is a rare clinical entity. The mechanism involved in testicular ascent is still not understood completely. Spasticity of cremaster muscle may cause secondary ascent of testis. The authors present 3 brothers with Pelizaeus Merzbacher syndrome, a rare, x-linked leukodystrophy in whom the testes bilaterally ascended from the normal scrotal position to an undescended position after onset of spasticity. PMID- 15547846 TI - An unusual cause of complete distal ureteral obstruction: giant fibroepithelial polyp. AB - Fibroepithelial polyp of the ureter is a rare benign neoplasm of mesodermal origin. It is an extremely rare cause of hydronephrosis in children. It usually causes partial ureteral obstruction without loss of renal function. The preferred treatment is endoscopic or surgical resection of the polyp with preservation of the renal unit. The authors present an adolescent patient with a nonfunctioning left hydronephrotic kidney caused by complete ureteral obstruction caused by a giant fibroepithelial polyp of the distal ureter. This is an extremely rare presentation and outcome of this benign ureteral neoplasm with resultant loss of renal unit. PMID- 15547872 TI - Improved survival outcome for hepatoblastoma. PMID- 15547873 TI - Outcome and complications after resection of hepatoblastoma. PMID- 15547874 TI - [Hepatitis C virus and renal transplantation]. AB - Liver disease has emerged as an important cause of morbidity and mortality after renal transplantation (RT). Hepatitis C virus (HCV) is the leading cause of liver disease after RT. The impact of HCV infection on patient and graft survival is currently a major concern. Retrospective studies with appropriate follow-up have mainly demonstrated that HCV positive patients have greater mortality compared to HCV negative recipients after RT. Novel investigations by large databases (United States Renal Data Systems (USRDS)) have shown that recipients of donor HCV positive kidneys are at an independently increased risk of mortality, adjusted hazard ratio 2.12 (95% confidence interval (95% CI), 1.72-2.87, p<0.001); there was no evidence that any subgroup was less affected. With appropriate informed consent, the use of a renal graft from an HCV positive donor could be offered to an HCV infected recipient. Many renal transplant candidates have satisfactory virological responses to antiviral therapy; the persistence of HCV clearance over a prolonged follow-up after RT has been recently noted. Further prospective studies are needed to define better the course of HCV infection among renal allograft recipients. PMID- 15547875 TI - [Hemodialysis access recirculation]. AB - Vascular access recirculation (R) allows the evaluation of the adequacy of the extracorporeal blood circuit in dialysis patients. The test verifies the correct needle position in patients with arterovenous fistulae (AVF) and the effective function of central venous catheters. In clinically uncomplicated native fistulae, a normal R test could avoid more complex procedures like blood flow measure or angiography. The AVF recirculation has two components, vascular access recirculation (AR) and cardiopulmonary recirculation (CPR). While the first phenomenon is well known, the second remained undetected for many years resulting in wrong R calculations with false positives. Using the correct formula, the great majority of AVF resulted in zero recirculation. The presence of R reduces the dialysis efficiency to critical levels, mainly in unsuspected cases. Among the numerous available R tests, the urea test is the oldest and historically the most commonly used method, but unfortunately it is labor intensive, with low sensitivity and specificity and with delayed results. The "ultrasound dilution"method is considered the gold standard, easy to perform, with good repeatability, but it is expensive requiring a specific device. Finally, the glucose infusion test (GIT) is a new low-cost test with immediate results and a very low detection limit, with good repeatability and high specificity and sensitivity. PMID- 15547876 TI - [Anti-cytokine therapy: present status and future perspectives in nephrology]. AB - The increasing understanding of the role of cytokines in chronic inflammatory disease, autoimmunity and neoplastic disease has led to a new generation of therapeutic agents, the anti-cytokine blocking agents. In this article, we review current knowledge of two different available approaches: the use of Thalidomide and the anti-cytokine antibody immune therapy. Thalidomide is an immunodulatory and antiangiogenic drug; the most pronounced effect of this drug is the inhibition of tumor necrosis factor-alpha (TNF-alpha ) production. A few years after its withdrawal from the European and Canadian markets due to severe teratogenic effects, the unexpected activity of Thalidomide in reactive lepromatous leprosy stimulated further study. After some confirmatory placebo controlled trials, multiple researches are now in progress to evaluate the optimal dose of Thalidomide in several autoimmune and neoplastic diseases. Both passive and active immunization can safely, transiently and effectively be used, as documented by animal experimentation and confirmed by clinical trials. Novel anti-cytokine therapeutic compounds, based on passive antibody immunization, are now available to treat rheumatoid arthritis and have been shown to help in Crohn's disease and in several other autoimmune diseases, and to control neoangiogenesis in cancer patients. The durability of the benefit, safety and pharmacoeconomic issues will determine whether this early success will prove to be a major breakthrough in the treatment of these painful and incurable diseases and eventually of other chronic inflammatory conditions in uremic patients. PMID- 15547877 TI - [Evidence-based guidelines in nephrology]. AB - INTRODUCTION: The preparation of evidence-based guidelines by the Nephrology Societies is fundamental to improve long-term outcomes of patients with chronic kidney diseases. However, this is a complex process and requires the interaction of clinicians and experts in epidemiology methods, and researchers and research enterprises. METHODS: In this review, we present the potential structure of a body for the coordination and development of evidence-based guidelines in a nephrology society and we address the major problems that can arise in this process describing strategies that could be used to overcome them. RESULTS: The development of evidence-based nephrology guidelines requires a structure; this should consist of a coordinating center and a number of working groups. The working groups is to identify specific research questions and to develop and synthetize the evidence in answer to the questions proposed. This shall be done in collaboration with the coordinating center. Draft guidelines produced by this process should be peer reviewed, disseminated and implemented. CONCLUSIONS: The development of evidence-based nephrology guidelines is a challenge for individual nephrology societies. These guidelines are different from typical research publications in that their success does not lie in the final publication, but in the actual dissemination and implementation, which is in the improvement of patient outcomes and its measurement. PMID- 15547878 TI - [Patient's consent to central venous catheterization]. AB - The need to obtain a patient's consent for his health care is a principle set out in the Italian Constitution, which safeguards a person's right to health. Articles 13 and 32.2 confirm a person's freedom and the right to make free decisions about one's medical treatment. Nobody must be obliged to any medical procedure, unless as by law enacted. The obligation to inform patients is important during the contractual phase: consent is an essential element in the professional contract governing the relationship between a physician and a patient. The former is obligated to inform the latter about his medical intervention clearly and precisely, to enable the patient to decide freely whether to undergo a medical procedure. At this point, it is also essential to obtain a patient's consent for those treatments that although they are carried out in a correct and careful way, could damage a person's physical integrity. The failure to obtain consent could give rise to a burden of responsibility on behalf of the clinician. A central venous catheterization in hemodialysis (HD) is a common procedure performed during routine nephrological treatments. Our signed informed consent form prior to introducing a central venous catheter is thought to satisfy requirements provided for in current regulations to give correct information. PMID- 15547879 TI - [An unexpected bottleneck]. AB - This is a clinical case concerning an uremic female patient who presented with an important oedema in the omolateral left arm, after the creation of an AV fistula. All the diagnostic procedures including the ultrasound color-Doppler were irrelevant. The patient was positive for a subclavian venous stenosis which was completely unexpected. This clinical case questions how useful it is in indirectly evaluating flow meters by ultrasound color-Doppler in the work-up of central venous steno-occlusions. PMID- 15547880 TI - [From the Mailing List of the Italian Society of Nephrology: hepatitis B prevention in dialysis units]. AB - The Mailing List SociSIN (ML-SIN) continues to develop, in addition to more experienced organizational topics, the discussion on clinical arguments. In the course of previous months, a message asking the opinions of expert Colleagues concerning preventative measures for HBsAg positive patients, in particular the use of separate rooms, has stimulated an extensive debate on this topic. This issue of the review is dedicated to the ML-SIN. It presents a short introduction dedicated to the definition of this problem. Thereafter, the messages regarding this argument are summarized, and, finally, the opinion of two experts regarding this matter is reported, experts identified as holding in account the importance of it international scientific contribution in on this specific discussion. PMID- 15547881 TI - [HCV-related liver disease in hemodialysis population: clinical and biochemical characteristic]. AB - Hepatitis C virus (HCV) infection remains frequent among patients on maintenance dialysis. It has been claimed that infrequent and slight abnormalities in serum aminotransferase activity could occur in dialysis patients with HCV. We describe a 61-year-old male patient on maintenance dialysis who acquired HCV by a nosocomial route. The natural history of HCV in this patient over 8 yrs featured frequent and high increases in serum aminotransferase and gamma-glutamyl transpeptidase (gamma-GT) levels. In December 2001, serum GOT and GPT were, respectively, 965 and 1294 UI/L; gamma-GT activity was 241 UI/L. HCV genotype was 2a/2c; median HCV RNA values in serum were 2.3 x 105 UI/mL (range, 1.14 x 104 to 4.6 x 105 UI/mL). Total bilirubin, serum albumin, and colinesterase levels remained normal over the entire follow-up. Liver biopsy was not performed and interferon (IFN) therapy was not given. Currently, biochemical liver tests (GOT/GPT/gamma-GT) are in the upper range of normal values and the patient remains viremic. Efficacy and tolerability of initial monotherapy with IFN for chronic hepatitis C among dialysis patients are briefly discussed. Further studies are warranted to define the optimal anti-viral regimen for chronic hepatitis C in the dialysis population. PMID- 15547882 TI - Laparoscopic skills training and assessment. AB - BACKGROUND: The introduction of laparoscopic techniques to general surgery was associated with many unnecessary complications, which led to the development of skills laboratories to train novice laparoscopic surgeons. This article reviews the tools currently available for training and assessment in laparoscopic surgery. METHODS: Medline searches were performed to identify articles with combinations of the following key words: laparoscopy, training, curriculum, virtual reality and assessment. Further articles were obtained by manually searching the reference lists of identified papers. RESULTS: Current training involves the use of box trainers with either innate models or animal tissues; it lacks objective assessment of skill acquisition. Virtual reality simulators have the ability to teach laparoscopic psychomotor skills, and objective assessment is now possible using dexterity-based and video analysis systems. CONCLUSION: The tools are now available for the development of a structured, competency-based, laparoscopic surgical training programme. PMID- 15547883 TI - Surgical safety and overwork. PMID- 15547884 TI - Prospective study of routine contrast radiology after total gastrectomy (Br J Surg 2004; 91: 1015-1019). PMID- 15547885 TI - Intra-abdominal hypertension and the abdominal compartment syndrome (Br J Surg 2004; 91: 1102-1110). PMID- 15547886 TI - Surgical management of primary anorectal melanoma (Br J Surg 2004; 91: 1183- 1187). PMID- 15547887 TI - Intraperitoneal and intraluminal microdialysis in the detection of experimental regional intestinal ischaemia (Br J Surg 2004; 91: 855-861). PMID- 15547888 TI - Mortality and morbidity of planned relaparotomy versus relaparotomy on demand for secondary peritonitis (Br J Surg 2004; 91: 1046-1054). PMID- 15547889 TI - Bowel injury as a complication of laparoscopy (Br J Surg 2004; 91: 1253-1258). PMID- 15547890 TI - Meta-analysis of short-term outcomes after laparoscopic resection for colorectal cancer (Br J Surg 2004; 91: 1111-1124). PMID- 15547894 TI - J. T. Hynes receives Hildebrand award / Cope scholar award to R. F. Ismagilov / P. Knochel receives cope scholar award. PMID- 15547891 TI - Systematic review of genetic influences on the prognosis of colorectal cancer (Br J Surg 2004; 91: 1275-1291). PMID- 15547895 TI - Supramolecular, bifurcated N-HOC-M bonding explains unusually low nu(CO) frequencies in metal carbonyl compounds: a case study. PMID- 15547896 TI - Individual molecules of dye-labeled DNA act as a reversible two-color switch upon application of an electric field. PMID- 15547897 TI - Self-construction of hollow SnO(2) octahedra based on two-dimensional aggregation of nanocrystallites. PMID- 15547898 TI - Quaternary ammonium groups in silica-associated proteins. PMID- 15547899 TI - A palladium(II)-clipped aromatic sandwich. PMID- 15547900 TI - [(Tp)(8)(H(2)O)(6)Cu(II) (6)Fe(III) (8)(CN)(24)](4+): a cyanide-bridged face centered-cubic cluster with single-molecule-magnet behavior. PMID- 15547901 TI - Surface-cap-mediated host-guest chemistry of semiconductor CdS: intercalative cation accumulation around a phenyl-capped CdS cluster and its notable effects on the cluster photoluminescence. PMID- 15547902 TI - Stereochemical assignment in acyclic lipids across long distance by circular dichroism: absolute stereochemistry of the aglycone of caminoside A. PMID- 15547903 TI - Enantioselective total synthesis of (+)-milnamide A and evidence of its autoxidation to (+)-milnamide D. PMID- 15547904 TI - Biofunctionalization of fluorescent rare-earth-doped lanthanum phosphate colloidal nanoparticles. PMID- 15547905 TI - Self-oxidation of a phenolate complex to a bimetallic stilbene quinone. PMID- 15547906 TI - alpha,alpha-Difluoro-H-phosphinates: useful intermediates for a variety of phosphate isosteres. PMID- 15547907 TI - A simple, one-step procedure for the formation of chiral metallamacrocycles. PMID- 15547908 TI - Readily available biaryl P,N ligands for asymmetric catalysis. PMID- 15547909 TI - Profiling kinase activities by using a peptide chip and mass spectrometry. PMID- 15547910 TI - A highly stereoselective addition of the anion derived from alpha-diazoacetamide to aromatic N-tosylimines. PMID- 15547911 TI - Highly ordered mesoporous bioactive glasses with superior in vitro bone-forming bioactivities. PMID- 15547912 TI - C(1)-symmetric sulfoximines as ligands in copper-catalyzed asymmetric Mukaiyama type aldol reactions. PMID- 15547913 TI - A mesoporous/crystalline composite material containing tin phosphate for use as the anode in lithium-ion batteries. PMID- 15547914 TI - Enantioselective total synthesis of the highly oxygenated 1,10-seco eudesmanolides eriolanin and eriolangin. PMID- 15547916 TI - Novel three-component reactions based on a Heck carbopalladation/cyclization domino reaction. PMID- 15547915 TI - [Yb(3)N(dpa)(6)][Yb(dpa)(3)]: a molecular nitride of a rare-earth metal with a Yb(3)N unit. PMID- 15547918 TI - Epstein-Barr virus associated post-transplantation lymphoproliferative disorder with hemophagocytosis in a child with Wiskott-Aldrich syndrome. AB - A 23-month-old boy with Wiskott-Aldrich syndrome (WAS) received human leukocyte antigen (HLA)-one locus mismatched, unmanipulated allogeneic bone marrow graft from his mother. An Epstein-Barr virus (EBV)-associated post-transplantation lymphoproliferative disorder (PTLD) of donor cell origin and hemophagocytosis syndrome with fever, lymphadenopathy, hepatosplenomegaly, seizures, involuntary movements and pancytopenia developed 52 days after transplantation. It was difficult to decide on the treatment strategy because the patient presented with B-cell hyperplasia that morphologically appeared malignant but was oligoclonal by heavy-chain analyses. Despite of donor leukocyte transfusion, low dose chemotherapy, and anti-B cell monoclonal antibody immunotherapy, the patient died 107 days after transplantation. PMID- 15547919 TI - Positive association between congenital anomalies and risk of neuroblastoma. AB - BACKGROUND: Case reports and epidemiological studies have suggested a relationship between congenital anomalies and childhood cancer, but some potential associations remain inconsistent. In this study, we investigated the association between congenital anomalies and neuroblastoma. PROCEDURE: We used data of a case-control study on neuroblastoma conducted from 1992 to 1994, including 538 children aged 0-19 years with newly diagnosed, histologically confirmed neuroblastoma and 504 controls identified by telephone random-digit dialing and matched to cases on date of birth. Information on congenital anomalies and potential confounding factors was collected through maternal telephone interviews using a structured questionnaire. We estimated odds ratios (OR) and 95% confidence intervals (CI), adjusted for reference age at diagnosis, mother's educational level, mother's race, and household income at birth. RESULTS: An association between the maternal report of any congenital anomalies and neuroblastoma (OR = 2.58; CI = 1.57-4.25) was observed. Neuroblastoma risk increased with increasing number of anomalies per child (OR = 3.90, CI = 1.27 11.9 for two anomalies or more), and when we restricted analyses to major anomalies (OR = 7.53, CI = 2.23-25.5). Genitourinary anomalies (OR = 5.84, CI = 1.67-20.4) and cardiac anomalies (OR = 4.27, CI = 1.22-15.0) had an elevated, but imprecise neuroblastoma risk. CONCLUSIONS: Our findings support the hypothesis of an association between neuroblastoma and congenital, especially urogenital and cardiac, anomalies. PMID- 15547920 TI - Cellular ubiquitination and proteasomal functions positively modulate mammalian nucleotide excision repair. AB - The ubiquitin-proteasome pathway is fundamental to synchronized continuation of many cellular processes, for example, cell-cycle progression, stress response, and cell differentiation. Recent studies have shown that the ubiquitin-proteasome pathway functions in the regulation of nucleotide excision repair (NER) in yeast. In order to investigate the role of the ubiquitin-proteasome pathway in the NER of mammalian cells, global genomic repair (GGR), and transcription-coupled repair (TCR) were examined in a mouse ts20 cell line that harbors a temperature sensitive ubiquitin-activating enzyme (E1). We found that E1 inactivation-induced ubiquitination deficiency decreased both GGR and TCR, indicating that the ubiquitination system is involved in the optimization of entire NER machinery in mammalian cells. We specifically inhibited the function of 19S proteasome subunit by overexpressing 19S regulatory complex hSug1 or its mutant protein hSug1mk in repair competent human fibroblast, OSU-2, cells and compared their capacity for NER. The results showed that 19S regulatory complex positively modulates NER in cells. In addition, we treated OSU-2 cells with the inhibitors of 20S subunit function, MG132 and lactacystin, and demonstrated that the catalytic activity of 20S subunit is also required for efficient NER. Moreover, the UV-induced recruitment of repair factor xeroderma pigmentosum protein C (XPC) to damage sites was negatively affected by treatment of repair competent cells with MG132. Taken together, we conclude that the ubiquitin-proteasome pathway has a positive regulatory role for optimal NER capacity in mammalian cells and appears to act through facilitating the recruitment of repair factors to DNA damage sites. PMID- 15547921 TI - SAGE profiling of UV-induced mouse skin squamous cell carcinomas, comparison with acute UV irradiation effects. AB - Ultraviolet (UV) irradiation is the primary environmental insult responsible for the development of most common skin cancers. To better understand the multiple molecular events that contribute to the development of UV-induced skin cancer, in a first study, serial analysis of gene expression (SAGE) was used to compare the global gene expression profiles of normal SKH-1 mice epidermis with that of UV induced squamous cell carcinomas (SCCs) from SKH-1 mice. More than 200 genes were found to be differentially expressed in SCCs compared to normal skin (P < 0.0005 level of significance). As expected, genes related to epidermal proliferation and differentiation were deregulated in SCCs relative to normal skin. However, various novel genes, not previously associated with skin carcinogenesis, were also identified as deregulated in SCCs. Northern blot analyses on various selected genes validated the SAGE findings: caspase-14 (reduced 8.5-fold in SCCs); cathepsins D and S (reduced 3-fold and increased 11.3-fold, respectively, in SCCs); decorin, glutathione S-transferase omega-1, hypoxia-inducible factor 1 alpha, insulin-like growth factor binding protein-7, and matrix metalloproteinase 13 (increased 18-, 12-, 12-, 18.3-, and 11-folds, respectively, in SCCs). Chemokine (C-C motif), ligand 27 (CCL27), which was found downregulated 12.7-fold in SCCs by SAGE, was also observed to be strongly downregulated 6-24 h after a single and multiple UV treatments. In a second independent study we compared the expression profile of UV-irradiated versus sham-treated SKH-1 epidermis. Interestingly, numerous genes determined to be deregulated 8 h after a single UV dose were also deregulated in SCCs. For instance, genes whose expression was upregulated both after acute UV-treated skin and SCCs included keratins 6 and 16, small proline-rich proteins, and S100 calcium binding protein A9. Studies like those described here do not only provide insights into genes and pathways involved in skin carcinogenesis but also allow us to identify early UV irradiation deregulated surrogate biomarkers of potential use in chemoprevention studies. PMID- 15547922 TI - The 2000 Burkitt lymphoma trial in Malawi. AB - BACKGROUND: We previously reported 57% 12-month event free survival (EFS) in Malawian children with stage I to III Burkitt lymphoma (BL) with an intermediate dose chemotherapy protocol lasting 77 days. This protocol was shortened to 42 days and evaluated in children with stage I to IV disease for EFS and toxicity. METHODS: All Malawian children admitted to Queen Elizabeth Central Hospital, from 03/08/2000 to 12/03/2002 with confirmed BL were eligible. A fine needle aspirate, bone marrow aspirate, cerebrospinal fluid cytology, haemoglobin (Hb), white cell count (WCC), malaria smear, ELISA for HIV, and abdominal ultrasound were performed routinely. Murphy staging was used. The first dose of chemotherapy (COP1) consisted of 300 mg cyclophosphamide (CPM), 1 mg vincristine, and 60 mg prednisone given on day 1 and followed by COP2 on day 8 (only for patients with larger tumour volumes, stage III or IV disease). The vincristine dose in COP2 was 2 mg. COMP1 and 2 given on days 22 and 36 consisted of 500 mg CPM, 2 mg vincristine, 60 mg prednisone, and 2 g methotrexate. All doses were calculated per body surface area. Intrathecal methotrexate and hydrocortisone were given with COP1 and 2. RESULTS: Forty-two patients, 30 boys and 12 girls median ages 6 and 7.5 years, respectively, had Murphy stage I(n5), II(n8), III(n21), and IV(n8) disease. The face was involved in 74%, abdomen in 55%, bone marrow in 14%, kidneys in 24%, and 12% had paraplegia. Fourteen children died during or shortly after completion of chemotherapy. Three of these were disease related. Twelve patients suffered a local relapse after 57-328 days, and one a CNS relapse at 76 days. The projected EFS at 12 months is 50% in stage I, 50% in stage II, 24% in stage III, 25% in stage IV, and 33% for all patients. The cumulative mean dose of CPM was 62 mg/kg in survivors and 64 mg/kg in children who relapsed. One third of patients experienced significant marrow suppression, and infections after COMP1. CONCLUSIONS: Thirty-three percent of children are in first remission at 12 months. The morbidity and mortality of treatment was high. The high relapse rate in all stages may be due to the low cumulative dose of CPM. PMID- 15547923 TI - Uncommon histiocytic disorders: the non-Langerhans cell histiocytoses. AB - BACKGROUND: Histiocytic disorders are currently identified by their component cells. The non-Langerhans Cell Histiocytoses (non-LCH) are a group of disorders defined by the accumulation of histiocytes that do not meet the phenotypic criteria for the diagnosis of Langerhans cells (LCs). The non-LCH consist of a long list of diverse disorders which have been difficult to categorize. A conceptual way to think of these disorders that make them less confusing and easier to remember is proposed based on immunophenotyping and clinical presentation. RESULTS: Clinically the non-LCH can be divided into 3 groups, those that predominantly affect skin, those that affect skin but have a major systemic component, and those that primarily involve extracutaneous sites, although skin may be involved. Immunohistochernically many of the non-LCH appear to arise from the same precursor cell namely the dermal dendrocyte. Juvenile Xanthogranuloma (JXG) is the model of the dermal dendrocyte-derived non-LCH. Other non-LCH with differing clinical presentation and occurring at different ages but with an identical immunophenotype appear to form a spectrum of the same disorder, deriving from the same precursor cell at different stages of maturation. They should be considered as members of a JXG family. Non-JXG family members include Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease). CONCLUSION: The non-LCH can be classified as JXG family and non-JXG family and subdivided according to fairly clear-cut clinical criteria. Utilization of this type of approach will allow better categorization, easier review of the literature and more accurate therapy decision-making. PMID- 15547924 TI - Intracranial germinoma: on whole-ventricular irradiation. AB - BACKGROUND: A current focus in the treatment of intracranial germinoma is on decreasing the volume of radiotherapy. We studied the theoretical impact of selecting different target volumes and treatment techniques. PROCEDURE: Retrospectively, five cases of localized germinoma were identified. Treatment plans were produced for three target volumes: involved field plans (i.e., targeting only gross disease), whole brain plans, and plans of increasing complexity targeting the ventricles (lateral-opposed, four-field and intensity modulated (IMRT)). A second series of patients was processed similarly except that no involved field plans were produced. RESULTS: When target volumes were compared, normal structures were substantially spared by ventricular irradiation but maximally spared by involved field radiation. When treatment techniques were assessed for ventricular irradiation, the greatest gain in tissue sparing occurred when switching from two to four fields. This gain was at the higher doses levels at the expense of an increased volume of tissue irradiated to lower doses. IMRT plans always spared a small additional amount of normal brain. Whole ventricular plans were compared to similar plans excluding the fourth ventricle. Whole brain dose was only improved by 4-6% by excluding the fourth ventricle. CONCLUSIONS: The substitution of whole-ventricular irradiation for whole-brain irradiation can spare a significant amount of normal tissue, thus providing a theoretical advantage with respect to the risk of late effects. Ventricular irradiation is best achieved with at least a four-field 3D configuration, and optimally through an IMRT plan. Exclusion of the fourth ventricle in the target volume had only a minimal impact on normal tissue doses. PMID- 15547925 TI - Massive intra-abdominal bleeding due to injury in the mesenteric artery during late breast reconstruction with a free TRAM flap: a case report. AB - We report on a rare case of major intra-abdominal bleeding that occurred in a patient undergoing late breast reconstruction with a transversus rectus abdominis musculocutaneous (TRAM) flap. During the flap-raising procedure, a small rupture was found at the base of the superior mesenteric artery, probably caused by electric current. The ruptured vessel required immediate laparotomy and suturing, and the final outcome of the patient was good. The massive bleeding, almost 8 fold as compared with three normal cases, caused clear activation of coagulation seen with elevated TAT values at the end of the operation, and remarkably enhanced fibrin turnover with elevated D-dimer, up to 25-fold from baseline, on the first postoperative day. As a result of major bleeding, the fibrinolytic system was also activated: a biphasic elevation of tPA was seen, first immediately after the bleeding event at 2 h, and later on the first postoperative day, followed by fibrinolytic shutdown associated with increased PAI-1 values. PMID- 15547926 TI - Usefulness of combined continuous suture and interrupted tie technique in hindlimb composite tissue allotransplantation in rats. AB - According to our experience of over 100 hindlimb transplantations, in rats, catching the posterior wall with an anterior wall suture is the most common cause of thrombosis and acute transplant failure. It is generally due to inadequate visibility of the lumen and consequent blind suture placement. With an intention to find a superior suture technique, a comparative study between standard end-to end interrupted and combined continuous sutures and interrupted tie techniques was performed. Twenty hindlimb transplantations to rats' femoral vessels were performed in each group. Anastomosis time, patency, complications, and animal and transplant survivals were compared. In the interrupted suture group (group I), anastomosis failure was encountered in one vein, with 95% of immediate patency rate. A redo of the venous anastomosis was performed and successful. The combined suture technique (group II) had 100% patency rates without complications. The average time for arterial anastomosis in groups I and II was 8.5 and 8.9 min, and for venous anastomosis, 10.7 and 9.6 min, respectively. At 2 weeks, no differences were found between groups as related to transplant and animal survival. PMID- 15547927 TI - Electrophysiological studies of myoclonus. AB - As myoclonus is often associated with abnormally increased excitability of cortical structures, electrophysiological studies provide useful information for its diagnosis and classification, and about its generator mechanisms. The electroencephalogram-electromyogram polygraph reveals the most important information about the myoclonus of interest. Jerk-locked back-averaging and evoked potential studies combined with recording of the long-latency, long-loop reflexes are useful to investigate the pathophysiology of myoclonus further, especially that of cortical myoclonus. Recent advances in magnetoencephalography and transcranial magnetic stimulation have contributed significantly to the understanding of some of the cortical mechanisms underlying myoclonus. Elucidation of physiological mechanisms underlying myoclonus in individual patients is important for selecting the most appropriate treatment. PMID- 15547928 TI - Prevalence of Factor V Leiden and other thrombophilic traits among Cretan children with malignancy. AB - BACKGROUND: The prevalence of thrombophilic traits, which might further enhance the risk of thrombotic complications in children treated for cancer, varies significantly among different populations. OBJECTIVE: To evaluate the prevalence of common thrombophilic traits of the East Mediterranean Region, among native Cretan children treated for malignancy. METHODS: Blood samples were consecutively collected from 31 native Cretan children treated for acute lymphoblastic leukaemia (n = 19) or other malignancies (n = 12) over 3 years. A molecular diagnosis based on the presence of Factor V Leiden (FVL), as well as on PT G20210A and MTHFR C677T mutation (in 14 patients) using PCR was applied. Patients who had central venous catheters (n = 29) were treated with an intensified thromboprophylaxis protocol that had been previously established in our institution. RESULTS: The prevalence of the FVL mutation was 19.4% (95% CI = 5 32). The allele frequency is estimated at 11.3% (95% CI: 3.5-19.1) which is higher than that reported for the population of the mainland of Greece. The prevalence of the PT G20210A and MTHFR C677T mutation was 14.3 and 71.4%, respectively (corresponding allele frequencies 7.1 and 50%, respectively). Only one patient developed thrombosis, having although no thrombophilic trait. CONCLUSIONS: Thrombophilic traits were relatively common in this group of native Cretan children treated for malignancy. Thromboprophylaxis should be considered in Cretan children in the presence of known acquired risk factors for thrombosis, but a larger prospective to study is first needed. PMID- 15547929 TI - Adjuvant chemotherapy for the treatment of advanced pediatric nonrhabdomyosarcoma soft tissue sarcoma: the National Cancer Institute experience. AB - BACKGROUND: The survival of children and adolescents with advanced (unresectable or metastatic) nonrhabdomyosarcoma soft tissue sarcoma (NRSTS) is poor. In order to clarify the role of combining chemotherapy with aggressive local control using surgery and/or radiation, we reviewed our institutional experience with the treatment of advanced pediatric NRSTS. PROCEDURE: We reviewed the charts of all patients less than 21 years treated for an advanced NRSTS at the National Cancer Institute (NCI) between 1983 and 2003. Tumor pathology was confirmed and demographic, disease, and treatment data were abstracted. Survival was calculated using standard methods. RESULTS: Of the 25 patients who were treated over the study period, 15 had metastatic disease and 10 had unresectable or incompletely resected disease at presentation. Twenty-one patients received chemotherapy consisting of the combination of vincristine, doxorubicin, cyclophosphamide, ifosfamide, and etoposide, and the remaining 4 received regimens that included doxorubicin. Twenty patients (80%) had a complete (5/25) or partial (15/25) response after chemotherapy alone. After the combination of chemotherapy and local control, 14 patients (56%) had a complete response (CR). The estimated 5 year overall and event-free survival (EFS) for all patients was 0.50 (standard error = 0.11) and 0.34 (standard error = 0.10), respectively. CONCLUSIONS: The combination of chemotherapy with aggressive local control in this cohort of pediatric patients with advanced NRSTS yielded results comparable to those observed in patients with advanced sarcomas that are chemotherapy responsive. Prospective randomized trials are needed to quantify the contribution of chemotherapy and to determine the ideal regimen. PMID- 15547930 TI - Pediatric Hodgkins disease in Morocco. PMID- 15547931 TI - Early complications in children with acute lymphoblastic leukemia presenting with hyperleukocytosis. AB - BACKGROUND: The optimal management of childhood acute lymphoblastic leukemia (ALL) with hyperleukocytosis is unclear, largely because the risk of leukostasis related complications is poorly characterized. PROCEDURE: We reviewed the presenting characteristics, initial management, and frequency and type of complications in all children seen at St. Jude Children's Research Hospital with previously untreated ALL and an initial leukocyte count >200 x 10(9)/L. RESULTS: A total of 178 children, representing 8% of all children with ALL, had an initial leukocyte count >200 x 10(9)/L; 67 patients had a leukocyte count >400 x 10(9)/L. Sixteen patients (9%) had neurological complications with 12 of these patients experiencing symptoms at presentation. Four patients (2%), all with initial leukocyte counts >400 x 10(9)/L, suffered a CNS hemorrhage. Pulmonary leukostasis occurred in 11 patients (6%). The degree of hyperleukocytosis was significantly predictive of neurological (P = 0.006) and respiratory (P = 0.014) complications. The majority of complications occurred at presentation. Cytoreduction (94 patients) decreased the leukocyte count but delayed initiation of chemotherapy (P = 0.013). CONCLUSIONS: Serious leukostasis-related complications are relatively uncommon in childhood ALL and most occur at presentation. Their incidence increases in proportion to the leukocyte count. A large subset of cases can be managed successfully without cytoreduction. Cytoreduction may be considered for patients with leukocyte counts >400 x 10(9)/L or patients who have complications at presentation. PMID- 15547932 TI - Aminoglycosides block the Kv3.1 potassium channel and reduce the ability of inferior colliculus neurons to fire at high frequencies. AB - The Kv3.1 potassium channel is expressed at high levels in auditory nuclei and contributes to the ability of auditory neurons to fire at high frequencies. We have tested the effects of streptomycin, an agent that produces progressive hearing loss, on the firing properties of inferior colliculus neurons and on Kv3.1 currents in transfected cells. We found that in inferior colliculus neurons, intracellular streptomycin decreased the current density of a high threshold, noninactivating outward current and reduced the rate of repolarization of action potentials and the ability of these neurons to fire at high frequencies. Furthermore, potassium current in CHO cells transfected with the Kv3.1 gene was reduced by 50% when cells were cultured in the presence of streptomycin or when streptomycin was introduced intracellularly in the pipette solution. In the presence of intracellular streptomycin, the activation rate of Kv3.1 current increased and inhibition by extracellular TEA become voltage dependent. The data indicate that streptomycin inhibits Kv3.1 currents by inducing a conformational change in the Kv3.1 channel. The hearing loss caused by aminoglycoside antibiotics may be partially mediated by their inhibition of Kv3.1 current in auditory neurons. PMID- 15547933 TI - Investigation of healthcare-associated transmission of Mycobacterium tuberculosis among patients with malignancies at three hospitals and at a residential facility. AB - BACKGROUND: Immunocompromised patients have an increased risk of experiencing progression of latent Mycobacterium tuberculosis infection (LTBI) to active tuberculosis (TB) disease. In January 2002, 2 patients with leukemia (Patients 1 and 2) developed pulmonary TB after recent exposure at 3 hospitals (Hospital A, Hospital B, and Hospital C) and at a residential facility for patients with cancer. Neither was known to have LTBI. Within 1 year, 3 other patients with malignancy and TB disease had been identified at these facilities, prompting an investigation of healthcare facility-associated transmission of M. tuberculosis. METHODS: The authors performed genotypic analysis of the five available M. tuberculosis isolates from patients with malignancies at these facilities, reviewed medical records, interviewed individuals who had identical M. tuberculosis genotypic patterns, and performed tuberculin skin testing (TST) and case finding for possible exposed contacts. RESULTS: Only Patients 1 and 2 had identical genotypic patterns. Neither patient had baseline TST results available. Patient 1 had clinical evidence of infectiousness 3 months before the diagnosis of TB was ascertained. Among employee contacts of Patient 1, TST conversions occurred in 1 of 59 (2%), 2 of 34 (6%), 2 of 32 (6%), and 0 of 8 who were tested at Hospitals A, B, and C and at the residential facility, respectively. Among the others who were exposed to Patient 1, 1 of 31 (3%), 1 of 30 (3%), 0 of 40 (0%), and 12 of 136 (9%) who were tested had positive TSTs at Hospitals A, B, and C and at the residential facility, respectively. CONCLUSIONS: Delayed TB diagnosis in 2 patients with leukemia resulted in the transmission of M. tuberculosis to 19 patients and staff at 3 hospitals and a residential facility. Baseline TB screening and earlier clinical recognition of active disease could reduce healthcare facility-associated transmission of M. tuberculosis among patients with malignancy. PMID- 15547934 TI - Expression of PKC in the developing zebrafish, Danio rerio. AB - Protein kinase C (PKC) is a family of enzymes involved in a wide range of biological functions. We investigated the expression of PKC-positive cells in zebrafish embryos and larvae within the first week of development to determine the developmental profile of PKC-containing cells. Our other goal was to determine if PKC alpha was associated with Rohon-Beard neurons during the first 5 days of development, when they are reported to undergo apoptosis. First, we confirmed the specificity of the antibodies by Western blotting zebrafish brain homogenates with anti-PKC and anti-PKC alpha, and detected single protein bands of approximately 78-82 kDa in size. Immunohistochemistry showed that several types of neurons were labeled, including neurons in the trigeminal ganglia, the dorsal spinal cord, and the dorsal root ganglia. Double-labeling with anti-PKC alpha and both anti-Islet-1 and zn12 confirmed the identity of the PKC-positive cells in the brain as trigeminal neurons, and in the spinal cord as Rohon-Beard cells. Some Rohon-Beard cells were labeled with anti-PKC alpha up to 7 days post fertilization (dpf). We performed TUNEL labeling and found no correlation between TUNEL-labeled and PKC alpha-labeled Rohon-Beard cells, suggesting that PKC alpha is not involved in Rohon-Beard apoptosis. Only approximately 40% of the approximately 130 Rohon-Beard cells at 24 h postfertilization (hpf) were positively labeled for PKC. Mauthner cells were labeled by anti-PKC, but not anti PKC alpha, suggesting that the major form of PKC within these cells was not PKC alpha. PMID- 15547935 TI - Centrophobism/thigmotaxis, a new role for the mushroom bodies in Drosophila. AB - Insects, like vertebrates, exhibit spatially complex locomotor activity patterns when foraging or navigating. Open field studies recently showed that Drosophila avoids central zones and stays at the periphery, an effect that can be interpreted as centrophobism and/or thigmotaxis. In this study, we further characterized this phenomenon and studied the responsible underlying neural mechanisms. The implication of the Drosophila mushroom bodies (MBs) in olfactory learning and memory processes is well documented. In an open field situation in which fly locomotor activity is recorded by video tracking, we show that center avoidance is greatly diminished in flies with hydroxyurea-ablated MBs, suggesting a new role for these structures. Furthermore, the temperature-sensitive allele of the dynamin gene shibire was expressed in various enhancer-trap P[GAL4] lines, disrupting synaptic transmission in different MB lobes. Specifically blocking the gamma lobes alters centrophobism/thigmotaxis while blocking the alpha/beta lobes does not, suggesting a functional specialization of MB lobes. Drosophila may serve as a new model system for elucidating the genetic and neural bases of such complex phenomena as centrophobism/thigmotaxis. PMID- 15547936 TI - CNS midline cells contribute to maintenance of the initial dorsoventral patterning of the Drosophila ventral neuroectoderm. AB - Dorsoventral patterning of the Drosophila ventral neuroectoderm is established by the expression of three evolutionarily conserved homeodomain genes: ventral nervous system defective (vnd), intermediate neuroblasts defective (ind), and muscle segment homeobox (msh) in the medial, intermediate, and lateral columns of the ventral neuroectoderm, respectively. It was not clear whether extrinsic factor(s) from the CNS midline cells influence the initial dorsoventral patterning by controlling the expression of the dorsoventral patterning genes. We show here that the CNS midline cells, specified by single-minded (sim), are essential for maintaining expression of the dorsoventral patterning genes. Ectopic expression of sim in the ventral neuroectoderm during the blastoderm stage repressed expression of the three homeodomain genes in the ventral neuroectoderm. This indicates that the identity of the CNS midline cells is established by a series of repressions of the three homeodomain genes in the ventral neuroectoderm. Ectopic expression of sim in the ventral neuroectoderm during initial neurogenesis induced ectopic ind expression in the medial column in addition to that in the intermediate column via EGFR signaling between the ventral neuroectoderm and midline cells. In contrast, it repressed the expression of vnd and msh in the medial and lateral columns, respectively. Our findings demonstrate that the CNS midline cells provide extrinsic positional information via EGFR signaling that maintains the initial subdivision of the ventral neuroectoderm into three dorsoventral columns during initial neurogenesis. PMID- 15547937 TI - Acute injections of brain-derived neurotrophic factor in a vocal premotor nucleus reversibly disrupt adult birdsong stability and trigger syllable deletion. AB - Behavioral variability serves an essential role in motor learning by enabling sensory feedback to select those motor patterns that minimize error. Birds use auditory feedback to learn how to sing, and their songs lose variability and become highly stereotyped, or crystallized, at the end of a sensitive period for sensorimotor learning. The molecular cues that regulate song variability are not well understood. In other systems, neurotrophins, and brain-derived neurotrophic factor (BDNF) in particular, can mediate various forms of neural plasticity, including sensitive period neural circuit plasticity and activity-dependent synapse formation, and may also influence learning and memory. Here, we have tested the hypothesis that neurotrophin expression in the robust nucleus of the arcopallium (RA), the telencephalic output controlling song, regulates song variability. BDNF and its receptor trkB are expressed in RA, and BDNF expression in RA appears to be highest in juveniles, when song is most variable and plastic, and synapse density highest. Thus, song variability and synaptic connectivity could be enhanced by augmented expression of BDNF in RA. In support of this idea, we found that BDNF injections into the adult RA induced the re-expression of juvenile-like phenotypes, including song variability and an increased synaptic density in RA. Furthermore, BDNF treatment also induced vocal plasticity, characterized by syllable deletions and persistent changes to the song patterns. These results suggest that endogenous BDNF could be a molecular regulator of the song variability essential to vocal plasticity and, ultimately, to song learning. PMID- 15547938 TI - Policy analysis of the use of Hepatitis B, Hemophilius influenzae type B, Steptococcus pneuomniae-conjugate and Rotavirus vaccines in the national immunization schedules. PMID- 15547939 TI - Molecular pathways involved in neural in vitro differentiation of marrow stromal stem cells. AB - In recent years several reports have claimed to demonstrate trans differentiation, namely that stem cells have been derived from a given tissue and have differentiated into phenotypes characteristic of different tissues following transplantation or in vitro treatment. For example, the mesenchymal stem cells, also referred to as marrow stromal stem cells (MSCs), present in bone marrow, have been induced to differentiate into neurons. We decided to investigate this phenomenon more in depth by a molecular and morphological follow-up. We analyzed the biochemical pathways that are currently induced to trigger neuron-like commitment and maturation of MSCs. Our studies suggest that: (i) the increase in cAMP, induced to differentiate MSCs, activates the classical PKA pathway and not through the exchange protein directly activated by cAMP (EPAC), a guanine nucleotide exchange factor for the small GTPase Rap1 and Rap2; (ii) MEK-ERK signaling could contribute to neural commitment and differentiation; (iii) CaM KII activity seems dispensable for neuron differentiation. On the contrary, its inhibition could contribute to rescuing differentiating cells from death. Our research also indicates that the currently used in vitro differentiation protocols, while they allow the early steps of neural differentiation to take place, are not able to further sustain this process. PMID- 15547940 TI - Charge distribution in the water molecule--a comparison of methods. AB - The charge distribution in the water molecule has been analyzed using a broad variety of basis sets, four different quantum mechanical methods (Hartree-Fock, Becke3LYP, MP2, and QCISD), and six population analysis methods (Mulliken, NPA, AIM, CHELPG, Merz-Kollman, and Resp). The influence of the molecular structure on the calculated atomic charges has been studied using small perturbations of the experimentally determined structure. PMID- 15547941 TI - Extensive theoretical studies on the low-lying electronic states of indium monochloride cation, InCl+. AB - The global potential energy curves for the 14 low-lying doublet and quartet Lambda-S states of InCl+ are calculated at the scalar relativistic MR-CISD+Q (multireference configuration interaction with single and double excitations, and Davidson's correction) level of theory. Spin-orbit coupling is accounted for via the state interaction approach with the full Breit-Pauli Hamiltonian, which leads to 30 Omega states. The computed spectroscopic constants of nine bound Lambda-S states and 17 bound Omega states are in good agreement with the available experimental data. The transition dipole moments and Franck-Condon factors of selected transitions are also calculated, from which the corresponding radiative lifetimes are derived. PMID- 15547942 TI - Arsenic trioxide represses constitutive activation of NF-kappaB and COX-2 expression in human acute myeloid leukemia, HL-60. AB - It has been proposed that eukaryotic nuclear factor nuclear factor kappa-B (NF kappaB) and cyclooxygenase-2 (COX-2) are implicated in the pathogenesis of many human diseases including cancer. Arsenic has been widely used in medicine in Oriental countries. Recent studies have shown that arsenic trioxide (As(2)O(3)) could induce in vitro growth inhibition and apoptosis of malignant lymphocytes, and myeloma cells. However, the molecular mechanisms by which As(2)O(3) initiates cellular signaling toward cell death are still unclear. In the present study, the effects of As(2)O(3) on NF-kappaB and COX-2 expression in HL-60 cells were investigated. As(2)O(3) suppressed DNA-binding activity of NF-kappaB composed of p65/p50 heterodimer through preventing the degradation of IkappaB-alpha and the nuclear translocation of p65 subsequently as well as interrupting the binding of NF-kappaB with their consensus sequences. Inhibitory effect of As(2)O(3) on NF kappaB DNA activity was dependent upon intracellular glutathione (GSH) and H(2)O(2) level, but not superoxide anion. Futhermore, we found that As(2)O(3) also downregulated the expression of COX-2, which has NF-kappaB binding site on its promoter through repressing the NF-kappaB DNA-binding activity. PMID- 15547943 TI - MEK partner 1 (MP1): regulation of oligomerization in MAP kinase signaling. AB - Specificity in signal transduction can be achieved through scaffolds, anchors, and adapters that assemble generic signal transduction components in specific combinations and locations. MEK Partner-1 (MP1) was identified as a potential "scaffold" protein for the mammalian extracellular signal-regulated kinase (ERK) pathway. To gain insight into the interactions of MP1 with the ERK pathway, we analyzed the ability of MP1 to bind to MEK1, ERK1, and to itself, and the regulation of these interactions. Gel filtration of cell lysates revealed two major MP1 peaks: a broad high molecular weight peak and a 28 kDa complex. An MP1 mutant that lost MEK1 binding no longer enhanced RasV12-stimulated ERK1 activity, and functioned as a dominant negative, consistent with the concept that MP1 function depends on facilitating these oligomerizations. Activation of the ERK pathway by serum or by RasV12 did not detectably affect MP1-MP1 dimerization or MP1-MEK1 interactions, but caused the dissociation of the MP1-ERK1 complex. Surprisingly, pharmacological inhibition of ERK activation did not restore the complex, suggesting that regulation of complex formation occurs independently of ERK phosphorylation. These results support the concept that MP1 functions as a regulator of MAP kinase signaling by binding to MEK1 and regulating its association with a larger signaling complex that may sequentially service multiple molecules of ERK. PMID- 15547945 TI - Role of Thy-1 in in vivo and in vitro neural development and regeneration of dorsal root ganglionic neurons. AB - We have examined the expression of Thy-1, an abundant glycosylphosphatidylinositol (GPI)-anchored glycoprotein, in dorsal root ganglia (DRG) and associated nerve fascicles, during postnatal development and following a nerve crush. The expression levels of Thy-1 in DRG neurons, dorsal roots, and central processes in spinal cord were rather low at postnatal day 2, and gradually increased as DRG neurons matured. During early development, the expression of Thy-1 within DRG neurons was low and equally distributed between plasma membrane and cytosol. With maturation, the staining intensities of Thy-1 in both the plasma membrane and the cytosol of DRG neurons became increased. We also studied Thy-1 expression in the regeneration of mature DRG neurons following the crush injury of sciatic nerve. Two days after the crush injury, Thy-1 expression dramatically decreased in the DRG neurons on the lesion side. Between 4 and 7 days after the injury, the expression of Thy-1 gradually increased and returned to a normal level 1 week after the sciatic nerve crush. The time course of the up-regulation of Thy-1 expression during regeneration matched that of the recovery of sensory functions, such as pain withdraw reflex, placing reflex, and the score of Basso-Beattie-Bresnahan Locomotor Rating Scale. Taken together, our results suggest that Thy-1 expression is developmentally regulated and is closely associated with the functional maturation of DRG neurons during both postnatal development and nerve regeneration. Furthermore, perturbation of Thy-1 function with anti-Thy-1 antibodies promoted neurite outgrowth from primary cultured DRG neurons, again confirming the inhibitory role of Thy-1 on neurite outgrowth. PMID- 15547947 TI - Formation of a physiological complex between TRPV2 and RGA protein promotes cell surface expression of TRPV2. AB - The transient receptor potential, sub-family Vanilloid (TRPV)(2) cation channel is activated in response to extreme temperature elevations in sensory neurons. However, TRPV2 is widely expressed in tissues with no sensory function, including cells of the immune system. Regulation of GRC, the murine homolog of TRPV2 has been studied in insulinoma cells and myocytes. GRC is activated in response to certain growth factors and neuropeptides, via a mechanism that involves regulated access of the channel to the plasma membrane. This is likely to be an important primary control mechanism for TRPV2 outside the CNS. Here, we report that a regulated trafficking step controls the access of TRPV2 to the cell surface in mast cells. In mast cells, elevations in cytosolic cAMP are sufficient to drive plasma membrane localization of TRPV2. We have previously proposed that the recombinase gene activator protein (RGA), a four-transmembrane domain, intracellular protein, associates with TRPV2 during the biosynthesis and early trafficking of the channel. We use a polyclonal antibody to RGA to confirm the formation of a physiological complex between RGA and TRPV2. Finally, we show that over-expression of the RGA protein potentiates the basal surface localization of TRPV2. We propose that trafficking and activation mechanisms intersect for TRPV2, and that cAMP mobilizing stimuli may regulate TRPV2 localization in non-sensory cells. RGA participates in the control of TRPV2 surface levels, and co-expression of RGA may be a key component of experimental systems that seek to study TRPV2 physiology. PMID- 15547948 TI - C3-induced 3LL cell proliferation is mediated by C kinase. AB - It has been demonstrated that the third component of complement (C3)(1) and its peptides increase normal and tumour cell proliferation. However, the signal cascade responsible for this phenomenon is still unknown. In this study, we elucidate some of the mechanisms involved in the signalling of C3 stimulation of cell proliferation. We have first investigated the in and out traffic of C3 peptides, then we have identified the subcellular localisation of internalised C3 and, finally, we have explored the role of protein phosphorylation in C3 traffic and in the proliferation of the Lewis lung carcinoma (3LL) cells. Our results indicate that traffic of C3 is not dependent on cytoskeletal integrity and requires protein kinase C-dependent phosphorylation. In addition, proliferation of 3LL cells stimulated by C3 depends on both C3 internalisation and protein kinase C phosphorylation. PMID- 15547949 TI - Evidence of a dominant negative mutant of yeast methionine aminopeptidase type 2 in Saccharomyces cerevisiae. AB - Eukaryotic methionine aminopeptidase type 2 (MetAP2, MetAP2 gene (MAP2)), together with eukaryotic MetAP1, cotranslationally hydrolyzes initiator methionine from nascent polypeptides when the side chain of the second residue is small and uncharged. In this report, we took advantage of the yeast (Saccharomyces cerevisiae) map1 null strain's reliance on MetAP2 activity for the growth and viability to provide evidence of the first dominant negative mutant of eukaryotic MetAP2. Replacement of the conserved His(174) with alanine within the C-terminal catalytic domain of yeast MetAP2 eliminated detectable catalytic activity against a peptide substrate in vitro. Overexpression of MetAP2 (H174A) under the strong GPD promoter in a yeast map1 null strain was lethal, whereas overexpression under the weaker GAL1 promoter slightly inhibited map1 null growth. Deletion mutants further revealed that the N-terminal region of MetAP2 (residues 2-57) is essential but not sufficient for MetAP2 (H174A) to fully interfere with map1 null growth. Together, these results indicate that catalytically inactive MetAP2 is a dominant negative mutant that requires its N terminal region to interfere with wild-type MetAP2 function. PMID- 15547950 TI - BMRP is a Bcl-2 binding protein that induces apoptosis. AB - Members of the Bcl-2 family of proteins play important roles in the regulation of cell death by apoptosis. The yeast Two-Hybrid system was utilized to identify a protein that interacts with the anti-apoptotic protein Bcl-2, designated BMRP. This protein corresponds to a previously known mitochondrial ribosomal protein (MRPL41). Binding experiments confirmed the interaction of BMRP to Bcl-2 in mammalian cells. Subcellular fractionation by differential centrifugation studies showed that both Bcl-2 and BMRP are localized to the same fractions (fractions that are rich in mitochondria). Northern blot analysis revealed a major bmrp mRNA band of approximately 0.8 kb in several human tissues. Additionally, a larger 2.2 kb mRNA species was also observed in some tissues. Western blot analysis showed that endogenous BMRP runs as a band of 16-17 kDa in SDS-PAGE. Overexpression of BMRP induced cell death in primary embryonic fibroblasts and NIH/3T3 cells. Transfection of BMRP showed similar effects to those observed by overexpression of the pro-apoptotic proteins Bax or Bad. BMRP-stimulated cell death was counteracted by co-expression of Bcl-2. The baculoviral caspase inhibitor p35 also protected cells from BMRP-induced cell death. These findings suggest that BMRP is a mitochondrial ribosomal protein involved in the regulation of cell death by apoptosis, probably affecting pathways mediated by Bcl-2 and caspases. PMID- 15547951 TI - Synergistic effect of prostaglandin F2alpha and cyclic AMP on glucose transport in 3T3-L1 adipocytes. AB - The combined effect of prostaglandin F2alpha (PGF2alpha) and cAMP on glucose transport in 3T3-L1 adipocytes was examined. In cells pretreated with PGF2alpha and 8-bromo cAMP for 8 h, a synergy between these two agents on glucose uptake was found. Insulin-stimulated glucose transport, on the other hand, was only slightly affected. The synergistic effect of these two agents was suppressed in the presence of cycloheximide and actinomycin D. In concord, immunoblot and Northern blot analyses revealed that GLUT1 protein and mRNA levels were both increased in cells pretreated with both PGF2alpha and 8-bromo cAMP, greater than the additive effect of each agent alone. The synergistic action of PGF2alpha with 8-bromo cAMP to enhance glucose transport was inhibited by GF109203X, a selective protein kinase C (PKC) inhibitor. In addition, in cells depleted of diacylglycerol-sensitive PKC by prolonged treatment with 4beta-phorbol 12beta myristate 13alpha-acetate, a PKC activator, the synergistic effects of PGF2alpha and 8-bromo cAMP on glucose transport and GLUT1 mRNA accumulation were both abolished. Taken together, these results indicate that PGF2alpha may act with cAMP in a synergistic way to increase glucose transport, probably through enhanced GLUT1 expression by a PKC-dependent mechanism. PMID- 15547952 TI - Semi-parametric and non-parametric methods for clinical trials with incomplete data. AB - Last observation carried forward (LOCF) and analysis using only data from subjects who complete a trial (Completers) are commonly used techniques for analysing data in clinical trials with incomplete data when the endpoint is change from baseline at last scheduled visit. We propose two alternative methods. The semi-parametric method, which cumulates changes observed between consecutive time points, is conceptually similar to the familiar life-table method and corresponding Kaplan-Meier estimation when the primary endpoint is time to event. A non-parametric analogue of LOCF is obtained by carrying forward, not the observed value, but the rank of the change from baseline at the last observation for each subject. We refer to this method as the LRCF method. Both procedures retain the simplicity of LOCF and Completers analyses and, like these methods, do not require data imputation or modelling assumptions. In the absence of any incomplete data they reduce to the usual two-sample tests. In simulations intended to reflect chronic diseases that one might encounter in practice, LOCF was observed to produce markedly biased estimates and markedly inflated type I error rates when censoring was unequal in the two treatment arms. These problems did not arise with the Completers, Cumulative Change, or LRCF methods. Cumulative Change and LRCF were more powerful than Completers, and the Cumulative Change test provided more efficient estimates than the Completers analysis, in all simulations. We conclude that the Cumulative Change and LRCF methods are preferable to LOCF and Completers analyses. Mixed model repeated measures (MMRM) performed similarly to Cumulative Change and LRCF and makes somewhat less restrictive assumptions about missingness mechanisms, so that it is also a reasonable alternative to LOCF and Completers analyses. PMID- 15547953 TI - Morphological irregularities and features of resistance to apoptosis in the dcp 1/pita double mutated egg chambers during Drosophila oogenesis. AB - In the present study, we demonstrate the most novel characteristic morphological features of Drosophila egg chambers lacking both dcp-1 and pita functions in the germline cells. Dcp-1 is an effector caspase and it has been previously shown to play an important role during Drosophila oogenesis [McCall and Steller, 1998 : Science 279 : 230-234; Laundrie et al., 2003 : Genetics 165 : 1881-1888; Peterson et al., 2003 : Dev Biol 260 : 113-123]. The completion of sequencing and annotation of the Drosophila genome has revealed that the dcp-1 gene is nested within an intron of another distinct gene, called pita, a member of the C2H2 zinc finger protein family that regulates transcriptional initiation. The dcp-1(-/ )/pita(-/-) nurse cells exhibit euchromatic nuclei (delay of apoptosis) during the late stages of oogenesis, as revealed by conventional light and electron microscopy. The phalloidin-FITC staining discloses significant defects in actin cytoskeleton arrangement. The actin bundles fail to organize properly and the distribution of actin filaments in the ring canals is changed compared to the wild type. The oocyte and the chorion structures have been also modified. The oocyte nucleus is out of position and the chorion appears to contain irregular foldings, while the respiratory filaments obtain an altered morphology. The dcp 1(-/-)/pita(-/-) egg chambers do not exhibit the rare events of spontaneously induced apoptosis, observed for the wild type flies, during mid-oogenesis. Interestingly, the mutated egg chambers are protected by staurosporine-induced apoptosis in a percentage of 40%, strongly suggesting the essential role of dcp-1 and/or pita during mid-oogenesis. PMID- 15547954 TI - Perspectives on the metallic interconnects for solid oxide fuel cells. AB - The various stages and progress in the development of interconnect materials for solid oxide fuel cells (SOFCs) over the last two decades are reviewed. The criteria for the application of materials as interconnects are highlighted. In terconnects based on lanthanum chromite ceramics demonstrate many inherent drawbacks and therefore are only useful for SOFCs operating around 1000 degrees C. The advance in the research of anode-supported flat SOFCs facilitates the replacement of ceramic interconnects with metallic ones due to their significantly lowered working temperature. Besides, interconnects made of metals or alloys offer many advantages as compared to their ceramic counterpart. The oxidation response and thermal expansion behaviors of various prospective metallic interconnects are examined and evaluated. The minimization of contact resistance to achieve desired and reliable stack performance during their projected lifetime still remains a highly challenging issue with metallic interconnects. Inexpensive coating materials and techniques may play a key role in pro-moting the commercialization of SOFC stack whose interconnects are constructed of some current commercially available alloys. Alternatively, development of new metallic materials that are capable of forming stable oxide scales with sluggish growth rate and sufficient electrical conductivity is called for. PMID- 15547955 TI - Solvothermal synthesis of nanosized CoSb(3) skutterudite. AB - Nanostructures enhance phonon scattering and improve the figure of merit of thermoelectric materials. Nanosized CoSb(3) skutterudite was synthesized by solvothermal methods using CoCl(2) and SbCl(3) as the precursors. A "two-step" model was suggested for the formation of CoSb(3) based on the X-ray diffraction analysis. The first step is the formation of cobalt diantimonide in the earlier stage during the synthesis process. Diantimonide was then combined with antimony atoms to form the skutterudite structured triantimonide, CoSb(3), in the later stage of the synthesis process as the second step. The synthesized CoSb(3) powders consist of irregular particles with sizes of about 20 nm and sheets of about 80 nm. PMID- 15547956 TI - Synthesis of arbutin by two-step reaction from glucose. AB - Arbutin was synthesized from glucose by two-step reaction below: (a) 2,3,4,6 tetra-O-acetyl-alpha-D-glucosyl chloride or bromide was prepared by glucose and acetyl halide (chloride or bromide). (b) 2,3,4,6-tetra-O-acetyl-alpha-D-glucosyl halide (Cl, Br) reacted with hydroquinone, methanol as solvent at pH=9.5-10.0. PMID- 15547957 TI - Synergetic effects for p-nitrophenol abatement using a combined activated carbon adsorption-electrooxidation process. AB - A novel fluidized electrochemical reactor that integrated advanced electrochemical oxidation with activated carbon (AC) fluidization in a single cell was developed to model pollutant p-nitrophenol (PNP) abatement. AC fluidization could enhance COD removal by 22%-30%. In such a combined process, synergetic effects on PNP and COD removal was found, with their removal rate being enhanced by 137.8% and 97.8%, respectively. AC could be electrochemically regenerated and reused, indicating the combined process would be promising for treatment of biorefractory organic pollutants. PMID- 15547958 TI - Self-desiccation mechanism of high-performance concrete. AB - Investigations on the effects of W/C ratio and silica fume on the autogenous shrinkage and internal relative humidity of high performance concrete (HPC), and analysis of the self-desiccation mechanisms of HPC showed that the autogenous shrinkage and internal relative humidity of HPC increases and decreases with the reduction of W/C respectively; and that these phenomena were amplified by the addition of silica fume. Theoretical analyses indicated that the reduction of RH in HPC was not due to shortage of water, but due to the fact that the evaporable water in HPC was not evaporated freely. The reduction of internal relative humidity or the so-called self-desiccation of HPC was chiefly caused by the increase in mole concentration of soluble ions in HPC and the reduction of pore size or the increase in the fraction of micro-pore water in the total evaporable water (T(r)/T(te) ratio). PMID- 15547959 TI - Preparation of natural alpha-tocopherol from non-alpha-tocopherols. AB - Non-alpha-tocopherols are hydroxymethylated and hydrogenated to produce alpha tocopherol in one pot process by simultaneously reacting with paraformaldehyde and hydrogen in the presence of catalysts of benzenesulfonic acid and 5% Pd/C in an autoclave. Effects of various operation conditions have been studied. The preferable reaction conditions are: temperature 180 degrees C to 200 degrees C, pressure 5.0 MPa, acid concentration 0.5 g/100 ml ethanol, mass ratio of Pd/C to tocopherols 7.1 g/100 g, and reaction time 5.0 h. A product with alpha-tocopherol content of 80% was obtained by using a raw material with a total tocopherols content of 80.54%. The conversion of non-alpha-tocopherols is almost 100%, and the mole yield of alpha-tocopherol is more than 90%. PMID- 15547960 TI - Comparison of volatile and semivolatile compounds from commercial cigarette by supercritical fluid extraction and simultaneous distillation extraction. AB - Supercritical carbon dioxide fluid extraction (SFE) was studied as a rapid method for extraction of volatile and semivolatile compounds of Chinese commercial cigarettes. The method was compared with simultaneous distillation and extraction (SDE). Temperature and pressure for the SFE were optimized. The extracts obtained by the two methods showed different characters in composition and represented differently the flavor characteristics of tobacco; compared to SDE, SFE can extract compounds within a shorter time and avoid the thermal degradation and solvent contamination of samples. The extracts by the two extraction methods are complementary for investigating the flavor characteristic of tobacco products. PMID- 15547961 TI - Land degradation, government subsidy, and smallholders' conservation decision: the case of the loess plateau in China. AB - Land degradation is one of the severe environmental problems in China. In order to combat land degradation, a soil conservation program was introduced since 2000 to reduce soil erosion by converting slope-cultivated land into forestry and pasture. This paper represents the first systematic attempt to investigate the impact of the soil conservation program on land degradation in the loess plateau. The results indicate that the soil conservation program to convert slope fields into forest or pasture is an effective way to combat soil erosion. However, a subsidy that is higher than profit of land use activity of slope fields before their conversion into forest and pasture is needed to encourage farmers to join the conservation program. A policy measure to encourage and assist farmers to develop sedentary livestock by using crops produced from fields as well as fodder and forage grass from the converted slope fields might contribute to combat soil erosion. Increase in off-farm job opportunities may encourage households to reduce cultivation in slope fields. That implies a policy measure to encourage rural urbanization might contribute to combat soil erosion. PMID- 15547962 TI - The mechanism of Fe (III)-catalyzed ozonation of phenol. AB - Fe (III)-catalyzed ozonation yielded better degradation rate and extent of COD (Chemical Oxygen Demand) or oxalic acid as compared with oxidation by ozone alone. Two parameters with strong effects on the efficiency of ozonation are pH of the solution and the catalyst (Fe(3+)) dosage. The existence of a critical pH value determining the catalysis of Fe (III) in acid conditions was observed in phenol and oxalic acid systems. The best efficiency of catalysis was obtained at a moderate concentration of the catalyst. A reasonable mechanism of Fe (III) catalyzed ozonation of phenol was obtained based on the results and literature. PMID- 15547963 TI - Synthesis of TiO(2) supported on activated carbon by MOCVD: operation parameters study. AB - The metallo-organic chemical vapor deposition (MOCVD) technique has been applied to the preparation of the photocatalyst titanium dioxide supported on activated carbon. The effects of various condition parameters such as carrier gas flow rate, source temperature and deposition temperature on the deposition rate were investigated. The maximum deposition rate of 8.2 mg/(g x h) was obtained under conditions of carrier gas flow rate of 400 ml/min, source temperature of 423 K and deposition temperature of 913 K. The deposition rate followed Arrhenius behavior at temperature of 753 K to 913 K, corresponding to activation energy E(a) of 51.09 kJ/mol. TiO(2) existed only in anatase phase when the deposition temperature was 773 K to 973 K. With increase of deposition temperature from 1073 K to 1273 K, the rutile content sharply increased from 7% to 70%. It was found that a deposition temperature of 773 K and a higher source temperature of 448 K resulted in finely dispersed TiO(2) particles, which were mainly in the range of 10-20 nm. PMID- 15547964 TI - Performance of PAHs emission from bituminous coal combustion. AB - Carcinogenic and mutagenic polycyclic aromatic hydrocarbons (PAHs) generated in coal combustion have caused great environmental health concern. Seventeen PAHs (16 high priority PAHs recommended by USEPA plus Benzo[e]pyrene) present in five raw bituminous coals and released during bituminous coal combustion were studied. The effects of combustion temperature, gas atmosphere, and chlorine content of raw coal on PAHs formation were investigated. Two additives (copper and cupric oxide) were added when the coal was burned. The results indicated that significant quantities of PAHs were produced from incomplete combustion of coal pyrolysis products at high temperature, and that temperature is an important causative factor of PAHs formation. PAHs concentrations decrease with the increase of chlorine content in oxygen or in nitrogen atmosphere. Copper and cupric oxide additives can promote PAHs formation (especially the multi-ring PAHs) during coal combustion. PMID- 15547965 TI - Experimental study on transient behavior of semi-open two-phase thermosyphon. AB - An experimental system was set up to measure the temperature, pressure, heat transfer rate and mass flow rate in a semi-open two-phase thermosyphon. The behaviors of a semi-open two-phase thermosyphon during startup, shutdown and lack of water were studied to get complete understanding of its thermal characteristics. The variation of wall temperature, heat-exchange condition and pressure fluctuations of semi-open two-phase thermosyphons showed that the startup of SOTPT needs about 60-70 min; the startup speed of SOTPT is determined by the startup speed of the condensation section; the average pressure in the heat pipe is equal to the environmental pressure usually; the shutdown of SOTPT needs about 30-50 min; a semi-open two-phase thermosyphon has good response to lack of water accident. PMID- 15547966 TI - Mutagenic effects of chromium trioxide on root tip cells of Vicia faba. AB - In this study on the mutagenic effects of different concentrations of chromium trioxide (CrO(3)) on Vicia faba root tip, micronucleus assay and chromosome aberration assay were used to determine the mitotic indexes, micronucleus aberration rate and chromosome aberration rate of Vicia faba root tip cells. The results showed that the effects of CrO(3) concentration on the mitotic indexes were complicated. CrO(3) increases the micronucleus aberration rate of Vicia faba root tip cells. It was found that within certain range of CrO(3) concentration the micronucleus aberration rate increased systematically with increased concentration of CrO(3), but that the micronucleus aberration rate decreased at higher level of CrO(3) and that CrO(3) also caused various types of chromosome aberration at a rate which increased systematically with increased concentration of CrO(3). We concluded that CrO(3) has significant mutagenic effect on Vicia faba root tip cells. PMID- 15547967 TI - EAG and behavioral responses of Helicoverpa armigera males to volatiles from poplar leaves and their combinations with sex pheromone. AB - Electroantennogram (EAG) evaluation of selected compounds from wilted leaves of black poplar, Populus nigra, showed that phenyl acetaldehyde, methyl salicylate, (E)-2-hexenal elicited strong responses from male antennae of Helicoverpa armigera. When mixed with sex pheromone (Ph), some volatiles, e.g. phenyl acetaldehyde, benzyl alcohol, phenylethanol, methylsalicylate, linalool, benzaldehyde, (Z)-3-hexenol, (Z)-3-hexenylacetate, (Z)-6-nonenol, cineole, (E)-2 hexenal, and geraniol elicited stronger responses from male antennae than Ph alone. Wind tunnel bioassay demonstrated that various volatiles could either enhance or inhibit the effect of synthetic sex pheromone. (E)-2-hexenal, (Z)-3 hexenol and linalool in combination with Ph could not induce any male to land on source at all, whereas phenyl acetaldehyde, benzaldehyde, (Z)-6-nonenol and salicylaldehyde combined with Ph enhanced male response rates by 58.63%, 50.33%, 51.85% and 127.78%, respectively, compared to Ph alone. These results suggested that some volatiles should modify sex pheromone caused behavior and that some of them could possibly be used as a tool for disrupting mating or for enhancing the effect of synthetic sex pheromone in the field. PMID- 15547968 TI - Effect of temperature on batch elastase production by Bacillus sp. EL31410. AB - The production of elastase by Bacillus sp. EL31410 at various temperatures was investigated. In order to study the effect of temperature on elastase fermentation, different cultivation temperatures, ranging from 39 degrees C to 28 degrees C, were evaluated in shake flask. The result indicated that 37 degrees C was best for cell growth at earlier stage; while maximum elastase activity was obtained when the cells were cultivated at 30 degrees C. This result was verified by batch fermentation in 5-L bioreactor under 37 degrees C and 30 degrees C temperature, respectively. The specific cell growth rate at 37 degrees C was higher than that at 30 degrees C during earlier stage of cultivation. The maximum value [5.5 U/(h x g DCW)] of elastase formation rate occurred at 24 h at 30 degrees C compared to 4.6 U/(h x g DCW) at 30 h at 37 degrees C. Based on these results, two-stage temperature shift strategy and oscillatory temperature cultivation mode were evaluated in the next study. When compared to single temperature of 37 degrees C or 30 degrees C, both two-stage temperature shift strategy and oscillatory temperature strategy improved biomass but did not yield the same result as expected for elastase production. The maximum biomass (both 8.6 g/L) was achieved at 30 h at 37 degrees C, but at 42 h using two-stage temperature cultivation strategy. The highest elastase production (652 U/ml) was observed at 30 degrees C in batch process. It was concluded that cultivation at constant temperature of 30 degrees C was appropriate for elastase production by Bacillus sp. EL31410. PMID- 15547969 TI - Scale-up of rifamycin B fermentation with Amycolatoposis mediterranei. AB - Study of the effect of dissolved oxygen and shear stress on rifamycin B fermentation with A. mediterranei XC 9-25 showed that rifamycin B fermentation with Amycolatoposis mediterranei XC 9-25 needs high dissolved oxygen and is not very sensitive to shearing stress. The scale-up of rifamycin B fermentation with A. mediterranei XC 9-25 from a shaking flask to a 15 L fermentor was realized by controlling the dissolved oxygen to above 25% of saturation in the fermentation process, and the potency of rifamycin B fermentation in the 15 L fermentor reached 10 g/L after 6-day batch fermentation. By continuously feeding glucose and ammonia in the fermentation process, the potency of rifamycin B fermentaion in the 15 L fermentor reached 18.67 g/L, which was 86.65% higher than that of batch fermentation. Based on the scale-up principle of constantly aerated agitation power per unit volume, the scale-up of rifamycin B fed-batch fermentation with continuous feed from a 15 L fermentor to a 7 m(3) fermentor and further to a 60 m(3) fermentor was realized successfully. The potency of rifamycin B fermentation in the 7 m(3) fermentor and in the 60 m(3) fermentor reached 17.25 g/L and 19.11 g/L, respectively. PMID- 15547970 TI - Ant colony system algorithm for the optimization of beer fermentation control. AB - Beer fermentation is a dynamic process that must be guided along a temperature profile to obtain the desired results. Ant colony system algorithm was applied to optimize the kinetic model of this process. During a fixed period of fermentation time, a series of different temperature profiles of the mixture were constructed. An optimal one was chosen at last. Optimal temperature profile maximized the final ethanol production and minimized the byproducts concentration and spoilage risk. The satisfactory results obtained did not require much computation effort. PMID- 15547971 TI - Research on the rheological properties of pesticide suspension concentrate. AB - This study reports research on pesticide suspension rheology and a new rheological parameter, the relative value of approach, which has great advantage for judging the physical stability of a pesticide suspension concentrate. Experiments showed that the system can form stable dispersions when the value of the relative value of approach (S(r)) is less than 0.1. PMID- 15547972 TI - Characterization of cellulose acetate micropore membrane immobilized acylase I. AB - This paper describes an innovative method for the immobilization of acylase I, which was entrapped into the CA-CTA micropore membrane. The most suitable casting solutions proportion for immobilizing the enzyme was obtained through orthogonal experiment. Properties of the enzyme membrane were investigated and compared with those of free enzyme and blank membrane. The thermal stability and pH stability of the enzyme inside the membrane were changed by immobilization. The optimum pH was found to be 6.0, which changes 1.0 unit compared with that of free acylase I. The optimum temperature was found to be about 90 degrees C, which is higher than that of free acylase I (60 degrees C). Experimental results showed that immobilization had effects on the kinetic parameters of acylase I. PMID- 15547973 TI - Mathematical modeling of salt-gradient ion-exchange simulated moving bed chromatography for protein separations. AB - The salt-gradient operation mode used in ion-exchange simulated moving bed chromatography (SMBC) can improve the efficiency of protein separations. A detailed model that takes into account any kind of adsorption/ion-exchange equilibrium, salt gradient, size exclusion, mass transfer resistance, and port periodic switching mechanism, was developed to simulate the complex dynamics. The model predictions were verified by the experimental data on upward and downward gradients for protein separations reported in the literature. All design and operating parameters (number, configuration, length and diameter of columns, particle size, switching period, flow rates of feed, raffinate, desorbent and extract, protein concentrations in feed, different salt concentrations in desorbent and feed) can be chosen correctly by numerical simulation. This model can facilitate the design, operation, optimization, control and scale-up of salt gradient ion-exchange SMBC for protein separations. PMID- 15547974 TI - Enhancing the quality metric of protein microarray image. AB - The novel method of improving the quality metric of protein microarray image presented in this paper reduces impulse noise by using an adaptive median filter that employs the switching scheme based on local statistics characters; and achieves the impulse detection by using the difference between the standard deviation of the pixels within the filter window and the current pixel of concern. It also uses a top-hat filter to correct the background variation. In order to decrease time consumption, the top-hat filter core is cross structure. The experimental results showed that, for a protein microarray image contaminated by impulse noise and with slow background variation, the new method can significantly increase the signal-to-noise ratio, correct the trends in the background, and enhance the flatness of the background, the consistency of the signal intensity. PMID- 15547975 TI - Parallel chemical genetic and genome-wide RNAi screens identify cytokinesis inhibitors and targets. AB - Cytokinesis involves temporally and spatially coordinated action of the cell cycle and cytoskeletal and membrane systems to achieve separation of daughter cells. To dissect cytokinesis mechanisms it would be useful to have a complete catalog of the proteins involved, and small molecule tools for specifically inhibiting them with tight temporal control. Finding active small molecules by cell-based screening entails the difficult step of identifying their targets. We performed parallel chemical genetic and genome-wide RNA interference screens in Drosophila cells, identifying 50 small molecule inhibitors of cytokinesis and 214 genes important for cytokinesis, including a new protein in the Aurora B pathway (Borr). By comparing small molecule and RNAi phenotypes, we identified a small molecule that inhibits the Aurora B kinase pathway. Our protein list provides a starting point for systematic dissection of cytokinesis, a direction that will be greatly facilitated by also having diverse small molecule inhibitors, which we have identified. Dissection of the Aurora B pathway, where we found a new gene and a specific small molecule inhibitor, should benefit particularly. Our study shows that parallel RNA interference and small molecule screening is a generally useful approach to identifying active small molecules and their target pathways. PMID- 15547976 TI - Oxygen-sensing mechanism of HemAT from Bacillus subtilis: a resonance Raman spectroscopic study. AB - Resonance Raman (RR) evidence for structural linkage between the distal side of heme pocket and the signaling domain of an oxygen sensing hemoprotein, HemAT-Bs, is reported. The band-fitting analyses of the RR spectra in the Fe-O2 stretching (nuFe-O2) region revealed the presence of three conformers with nuFe-O2 at 554, 566, and 572 cm-1, which reflect different H-bond strengths on the bound O2 molecule. While recent X-ray analysis for CN--bound HemAT-Bs suggested the importance of Thr95 and Tyr70, the species with the strongest H-bond (554 cm-1) was deleted in the T95A mutant and also by removal of the linker and signal domains; however, the Y70F mutant maintained the same three conformers. A scheme for specific O2 sensing and signaling mechanism is discussed. PMID- 15547977 TI - A slippery slope: mechanistic analysis of the intramolecular Scholl reaction of hexaphenylbenzene. AB - DFT calculations support an arenium cation-based mechanism for the Scholl reaction converting hexaphenylbenzene to hexa-peri-benzocoronene. The curve connecting fully benzenoid intermediates on the potential energy diagram is convex. This "slippery slope" provides an explanation for the ease of this cascade Scholl reaction. The calculated reaction coordinate predicts that intermediates will not accumulate; this prediction is verified by experiment. PMID- 15547978 TI - Molecular cube of Re(II) and Mn(II) that exhibits single-molecule magnetism. AB - A pseudocubic, paramagnetic cluster of ReII and MnII, [{MnCl}4{Re(triphos)(CN)3}4], has been prepared, and its magnetic properties have been investigated. Antiferromagnetic coupling is observed between the "S = 1/2" ReII and S = 5/2 MnII centers resulting in an effective S = 8 ground state. AC susceptibility studies reveal that the molecule is a single-molecule magnet with an effective barrier for magnetization reversal of Ueff = 8.8 cm-1. PMID- 15547979 TI - Structural rationalization of a large difference in RNA affinity despite a small difference in chemistry between two 2'-O-modified nucleic acid analogues. AB - Chemical modification of nucleic acids at the 2'-position of ribose has generated antisense oligonucleotides (AONs) with a range of desirable properties. Electron withdrawing substituents such as 2'-O-[2-(methoxy)ethyl] (MOE) confer enhanced RNA affinity relative to that of DNA by conformationally preorganizing an AON for pairing with the RNA target and by improving backbone hydration. 2'-Substitution of the ribose has also been shown to increase nuclease resistance and cellular uptake via changes in lipophilicity. Interestingly, incorporation of either 2'-O [2-(methylamino)-2-oxoethyl]- (NMA) or 2'-O-(N-methylcarbamate)-modified (NMC) residues into AONs has divergent effects on RNA affinity. Incorporation of 2'-O NMA-T considerably improves RNA affinity while incorporation of 2'-O-NMC-T drastically reduces RNA affinity. Crystal structures at high resolution of A-form DNA duplexes containing either 2'-O-NMA-T or 2'-O-NMC-T shed light on the structural origins of the surprisingly large difference in stability given the relatively minor difference in chemistry between NMA and NMC. NMA substituents adopt an extended conformation and use either their carbonyl oxygen or amino nitrogen to trap water molecules between phosphate group and sugar. The conformational properties of NMA and the observed hydration patterns are reminiscent of those found in the structures of 2'-O-MOE-modified RNA. Conversely, the carbonyl oxygen of NMC and O2 of T are in close contact, providing evidence that an unfavorable electrostatic interaction and the absence of a stable water structure are the main reasons for the loss in thermodynamic stability as a result of incorporation of 2'-O-NMC-modified residues. PMID- 15547980 TI - Optically active sum frequency generation from molecules with a chiral center: amino acids as model systems. AB - With amino acids as model systems, optically active sum frequency generation (OA SFG) was used to probe the chirality of molecules with a chiral center and an intrinsically achiral chromophore in isotropic solution for the first time. Like that of circular dichroism (CD), the OA-SFG's near electronic resonance originates from the extrachromophoric chiral perturbation on the carboxyl chromophore. The difference in the relative strengths of OA-SFG and CD among different amino acids can be explained by the difference in the details of perturbations. PMID- 15547981 TI - Electron transfer rates in DNA films as a function of tether length. AB - A homologous series of DNA-modified electrodes has been investigated in which the molecular tether length varies. Using intercalated, covalently bound daunomycin as a redox probe, an exponential dependence of electron transfer rates on the number of intervening methylene groups in the sigma-bonded tether is observed. In contrast, variation in DM position within DNA yields no detectable change in rate. These data confirm that overall electron transfer rates in DNA films are limited by the tether, not the DNA. PMID- 15547982 TI - Lipid-based nanotubes as functional architectures with embedded fluorescence and recognition capabilities. AB - A limited combinatorial strategy was used to synthesize a small library of soft lipid-based materials ranging from structurally unordered fibers to highly uniform nanotubes. The latter nanotubes are comprised of a bilayer structure with interdigitated alkyl chains associated through hydrophobic interactions. These tubes contain accessible 2,6-diaminopyridine linkers that can interact with thymidine and related nucleosides through multipoint hydrogen bonding, thereby quenching the intrinsic fluorescence of the aromatic linker. These results are the first example of a systematic strategy to design functional lipid nanotubes with precise structural and functional features. PMID- 15547983 TI - How many hydrogen atoms can be bound to a metal? Predicted MH12 species. AB - Quantum chemical calculations predict the existence of new molecular species with general formula MH12, where M is a group 6 atom. The previous MHn species had n values up to 9. The new systems with n = 12 would be a new record for metal hydrides. PMID- 15547984 TI - Dimeric molecular capsules under redox control. AB - A new tetraferrocenylurea calix[4]arene was prepared in order to investigate its dimerization properties. 1H NMR spectroscopic data clearly established that this calixarene forms stable dimeric molecular capsules in chloroform solution. The dimeric capsules are stabilized by the formation of multiple hydrogen bonds between the urea functional groups on the calixarene upper rims. In the dimer, eight ferrocene groups are held in proximity to the "seam" of hydrogen bonds. Upon oxidation of the ferrocene residues, electrochemical, PGSE NMR, and IR spectroscopic data revealed that the hydrogen bonds are broken, leading to the dissociation of the dimeric molecular capsule. PMID- 15547985 TI - General method for suppression of diagonal peaks in heteronuclear-edited NOESY spectroscopy. AB - A novel approach is proposed for suppression of diagonal peaks in 15N- and 13C edited NOESY spectroscopy based on subtracting a spectrum with only diagonal peaks from the conventional NOESY spectrum. This method can be applied to most heteronuclear-edited NOESY experiments. It is far more sensitive than the TROSY based approach for biomolecules with little TROSY effect, and nearly complete suppression of diagonal peaks can be achieved. The method has been demonstrated on samples of 15N-labeled calmodulin (17 kDa), 13C-labeled DdCAD-1 (24 kDa), and 13C-labeled hemoglobin (65 kDa), showing that cross-peaks very close to diagonals can be assigned reliably based on the difference spectra. PMID- 15547986 TI - Dendronized linear polymers via "click chemistry". AB - Dendronized linear polymers are prepared from dendritic azides and poly(vinylacetylene) using "click chemistry." The Cu(I)-catalyzed Huisgen [2 + 3] cycloaddition is quantitative up to the third generation. PMID- 15547987 TI - Fabrication of nanocables by electrochemical deposition inside metal nanotubes. AB - We report a novel route for fabricating Au-Te nanocables. Using nanoporous polycarbonate tract-etching (PCTE) membrane as the template, Au nanotubes were fabricated by electroless Au deposition inside the nanopores of the PCTE membrane. Using the Au nanotube membrane as a second template, Te was deposited on the surfaces of the Au nanotubes by slow electrochemical deposition, taking advantage of underpotential deposition (UPD). The deposition rate was sufficiently slow to radially grow Te nanotubes coaxially within the Au nanotubes to form nanocables. PMID- 15547988 TI - Time-dependent DFT study of [Fe(CN)6]4- sensitization of TiO2 nanoparticles. AB - Time-dependent DFT calculations have been performed on the absorption spectrum of [Fe(CN)6]4- adsorbed on a TiO2 anatase nanoparticle model to provide a detailed description of the electronic structure for this prototype system and to understand the character of the states involved in the molecule --> semiconductor charge transfer process. Our results show that a direct charge injection process from an occupied dye molecular state to a nanoparticle excited state localized on a few Ti atoms, rather than to a delocalized conduction state, effectively takes place in this system, in agreement with recent experimental evidence. PMID- 15547989 TI - In situ control of cellular growth and migration on substrates using microelectrodes. AB - We describe an electrochemical method to direct the growth and migration of mammalian cells on a substrate during cultivation in situ. Exposing the albumin coated substrate to an oxidizing agent, hypobromous acid, electrochemically generated at the tip of the scanning microelectrode, locally switched the substrate from cytophobic to cell-adhesive. This transformation generated the formation of cellular micropatterns. Since the concentration of the oxidizing agent required for the surface processing did not cause significant damage to the cell cultures, we were able to direct in situ cellular proliferation and migration by drawing adhesive micropatterns over the preexisting cellular pattern. PMID- 15547990 TI - Molecular recognition remolds the self-assembly of hydrogelators and increases the elasticity of the hydrogel by 10(6)-fold. AB - Addition of vancomycin (1, the receptor) to the supramolecular hydrogel of self assembled pyrene-d-Ala-d-Ala (2, the ligand) increases the storage modulus of the hydrogel of 2 by about 106-fold. Rheology, microscopy, and spectroscopy investigations suggest that the two-dimensional polymers, formed by the ligand receptor interaction between 1 and 2, and the self-dimerization of 1, are mainly responsible for the observed dramatic increase in elasticity. PMID- 15547991 TI - Metal-assisted assembly and stabilization of collagen-like triple helices. AB - Single-chain and TRIS-assembled collagen mimetic peptide structures incorporating catechol groups were synthesized. When 1/3 equiv of Fe3+ was added to the single chain compound in 50 mM CAPS buffer (pH 10), the 1:3 Fe3+-catechol complex that formed acted as an N-terminal scaffold to assemble the triple helix. When 1 equiv of Fe3+ was added to the TRIS-assembled compound in the buffer solution, the Fe3+ catechol complex acted as an extra C-terminal scaffold, which lead to a triple helix with both termini tethered. The formation of this C-terminal complex increased the Tm by a remarkable 22 degrees C! PMID- 15547992 TI - Li4C60: a polymeric fulleride with a two-dimensional architecture and mixed interfullerene bonding motifs. AB - All known fullerene polymers have interfullerene connections via either [2 + 2] cycloaddition or single C-C bonds. The high-resolution synchrotron X-ray powder diffraction technique was employed here to determine the crystal structure of the Li4C60 fulleride. We find that the ground state of Li4C60 is a two-dimensional polymer with monoclinic crystal symmetry and an unprecedented architecture, combining both the [2 + 2] cycloaddition and the single C-C bridging motifs. The small size of the Li+ cations is crucial in stabilizing the resulting tightly packed polymeric structure. PMID- 15547993 TI - Kinetic and thermodynamic preferences in aryl vs benzylic C-H bond activation with cationic Pt(II) complexes. AB - Anhydrous cationic Pt(II) complexes [(NN)Pt(CH3)(CF3CD2OD)]+ (1, NN = ArN=C(Me) C(Me)=NAr), which are obtained by reaction of (NN)Pt(CH3)2 with B(C6F5)3 in CF3CD2OD, activate C-H bonds of benzene and methylbenzenes, with enhanced reactivity compared to the previously prepared equilibrium mixtures with the (thermodynamically favored) aquo complexes. For methylbenzenes (toluene, p xylene, mesitylene), activation at the aromatic and benzylic positions are kinetically competitive, but the product of the latter is strongly favored thermodynamically. This unusual trend is attributed to formation of eta3-benzyl structures, which can be observed spectroscopically for 1,4-diethylbenzene activation. PMID- 15547994 TI - How large is the conjugative stabilization of diynes? AB - The conjugation stabilization energies of dienes and diynes are considerably larger than estimates based on heat of hydrogenation differences between 1,3 butadiyne and 1-butyne as well as between 1,3-butadiene and 1-butene. Such comparisons do not take into account the counterbalancing hyperconjugative stabilization of the partially hydrogenated products by their ethyl groups. When alkyl hyperconjugation is considered, the conjugation stabilization of diynes ( approximately 9.3 kcal/mol) is found by two methods (involving isomerization of nonconjugated into conjugated isomers and heats of hydrogenation) to be larger than that of dienes ( approximately 8.2 kcal/mol). PMID- 15547995 TI - Lewis acid-mediated selective chlorinations of silyl enolate. AB - A new method involving efficient, widely applicable, and highly selective alpha chlorination of simple silyl enolate with Lewis acid and an alpha,alpha-dichloro 1,3-dicarbonyl controller unit was reported. Diastereoselectivity and enantioselectivity of the reaction were investigated. High reactivity and selectivity were achieved by using alpha,alpha-dichlorinated malonic ester. PMID- 15547996 TI - An efficient synthesis of phthalocyanines based on an unprecedented double addition of oximes to phthalonitriles. AB - A novel and efficient method is reported for the preparation of phthalocyanines from phthalonitrile or its derivatives which utilizes low-cost reagents of hydrolytic stability, high simplicity, and easy accessibility, i.e., oximes, and proceeds via an unprecedented double addition of oximes to the nitrile group. PMID- 15547997 TI - Conversion from arenes having a benzene ring to those having a picolinic acid by simple growing cell reactions using Escherichia coli that expressed the six bacterial genes involved in biphenyl catabolism. AB - The comprehensive bioconversion of aromatic compounds with a benzene ring to a picolinic acid was achieved with a recombinant Escherichia coli strain that expressed the six genes involved in biphenyl catabolism, these being the bphA1(2072)A2A3A4 genes encoding the evolved biphenyl dioxygenase, the bphB gene encoding dihydrodiol dehydrogenase, and the bphC gene encoding catechol 2,3 dioxygenase. PMID- 15547998 TI - The enantioselective Tsuji allylation. AB - The first catalytic enantioselective examples of the Tsuji allylation using enol carbonates and enol silanes are described. The products possess a quaternary stereogenic center and are useful building blocks for synthetic chemistry. PMID- 15547999 TI - A strain-promoted [3 + 2] azide-alkyne cycloaddition for covalent modification of biomolecules in living systems. AB - Selective chemical reactions that are orthogonal to the diverse functionality of biological systems have become important tools in the field of chemical biology. Two notable examples are the Staudinger ligation of azides and phosphines and the Cu(I)-catalyzed [3 + 2] cycloaddition of azides and alkynes ("click chemistry"). The Staudinger ligation has sufficient biocompatibility for performance in living animals but suffers from phosphine oxidation and synthetic challenges. Click chemistry obviates the requirement of phosphines, but the Cu(I) catalyst is toxic to cells, thereby precluding in vivo applications. Here we present a strain promoted [3 + 2] cycloaddition between cyclooctynes and azides that proceeds under physiological conditions without the need for a catalyst. The utility of the reaction was demonstrated by selective modification of biomolecules in vitro and on living cells, with no apparent toxicity. PMID- 15548000 TI - Attaching organic semiconductors to gate oxides: in situ assembly of monolayer field effect transistors. AB - This study unveils a new tetracene derivative that forms dense, upright monolayers on the surface of aluminum oxide. These monolayers spontaneously self organize into the active layer in nanoscale field-effect transistor devices when aluminum oxide is used as the dielectric layer. This method gives high yields of working devices that have source-drain distances that are less than 60 nm, thereby providing a method to electrically probe the monolayer assemblies formed from approximately 10 zeptomoles of material (approximately 104 molecules). Moreover, this study delineates a new avenue for research in thin-film organic transistors where the active molecules are linked to the dielectric surface to form a monolayer transistor. PMID- 15548001 TI - Directed evolution of a glycosynthase via chemical complementation. AB - Recently, we reported a general assay for enzyme catalysis based on the yeast three-hybrid assay, Chemical Complementation, which is intended to expand the range of chemical reactions to which directed evolution can be applied. Here, Chemical Complementation was applied to a glycosynthase derived from a retaining glycosidase, an important class of enzymes for carbohydrate synthesis. Using the yeast three-hybrid assay, the glycosynthase activity of the E197A mutant of the Cel7B from Humicola insolens was linked to transcription of a LEU2 reporter gene, making cell growth dependent on glycosynthase activity in the absence of leucine. Then the LEU2 selection was used to isolate the most active glycosynthase from a Glu197 saturation library, yielding an E197S Cel7B variant with a 5-fold increase in glycosynthase activity. These results not only establish Chemical Complementation as a platform for the directed evolution of glycosynthases, but also show the generality of this approach and the ease with which it can be applied to new chemical reactions. PMID- 15548002 TI - Vanadium bromoperoxidase-catalyzed biosynthesis of halogenated marine natural products. AB - Marine red algae (Rhodophyta) are a rich source of bioactive halogenated natural products. The biogenesis of the cyclic halogenated terpene marine natural products, in particular, has attracted sustained interest in part because terpenes are the biogenic precursors of many bioactive metabolites. The first enzymatic asymmetric bromination and cyclization of a terpene, producing marine natural products isolated from red algae, is reported. Vanadium bromoperoxidase (V-BrPO) isolated from marine red algae (species of Laurencia, Plocamium, Corallina) catalyzes the bromination of the sesquiterpene (E)-(+)-nerolidol producing alpha-, beta-, and gamma-snyderol and (+)-3beta-bromo-8-epicaparrapi oxide. alpha-Snyderol, beta-snyderol, and (+)-3beta-bromo-8-epicaparrapi oxide have been isolated from Laurencia obtusa, and each have also been isolated from other species of marine red algae. gamma-Snyderol is a proposed intermediate in other bicyclo natural products. Single diastereomers of beta-snyderol, gamma snyderol, and mixed diastereomers of (+)-3beta-bromo-8-epicaparrapi oxide (de = 20-25%) are produced in the enzyme reaction, whereas two diastereomers of these compounds are formed in the synthesis with 2,4,4,6-tetrabromocyclohexa-2,5 dienone (TBCO). V-BrPO likely functions by catalyzing the two-electron oxidation of bromide ion by hydrogen peroxide producing a bromonium ion or equivalent in the active site that brominates one face of the terminal olefin of nerolidol. These results establish V-BrPO's role in the biosynthesis of brominated cyclic sesquiterpene structures from marine red algae for the first time. PMID- 15548003 TI - (S,S)-trans-cyclopentane-constrained peptide nucleic acids. a general backbone modification that improves binding affinity and sequence specificity. AB - Replacing the ethylenediamine portion of aminoethylglycine peptide nucleic acids (aegPNAs) with one or more (S,S)-trans-cyclopentane diamine units significantly increases binding affinity and sequence specificity to complementary DNA, making these modified PNAs ideal for use as nucleic acid probes in genomic analysis. The synthesis and study of this new class of PNAs (tcypPNAs) is described in which trans-cyclopentane diamine has been incorporated into several positions, and in varying number, within PNA backbones of mixed-base sequences. PMID- 15548004 TI - Ring closing enyne metathesis: control over mode selectivity and stereoselectivity. AB - Ring closing enyne metathesis to form 10-15-membered rings was achieved by using a tartrate-derived linker to attach ene and yne subunits. The exo/endo selectivity of the ring closure reaction of these substrates was found to be a function of ring size, whereby larger rings (12-15) give endo-products selectively, while smaller rings (5-11) give exo-products. The E/Z selectivity of the resultant macrocyclic 1,3-dienes was not predictable except for 10- and 11 membered rings. However, both the exo/endo-mode selectivity of the ring closure and the E/Z selectivity of the 1,3-dienes were improved by performing these reactions under ethylene atmosphere. The presence of ethylene induces a selective cross metathesis between the alkyne moiety and ethylene to generate an acyclic 1,3-diene which can undergo ring closing diene metathesis between the isolated olefin and the distal monosubstituted double bond of the 1,3-diene to generate exclusively the endo-product with high E-selectivity. PMID- 15548005 TI - Direct chemical synthesis of the beta-mannans: linear and block syntheses of the alternating beta-(1-->3)-beta-(1-->4)-mannan common to Rhodotorula glutinis, Rhodotorula mucilaginosa, and Leptospira biflexa. AB - Two stereocontrolled syntheses of a methyl glycoside of an alternating beta-(1- >4)-beta-(1-->3)-mannohexaose, representative of the mannan from Rhodotorula glutinis, Rhodotorula mucilaginosa, and Leptospira biflexa, are described. Both syntheses employ a combination of 4,6-O-benzylidene- and 4,6-O-p methoxybenzylidene acetal-protected donors to achieve stereocontrolled formation of the beta-mannoside linkage. The first synthesis is a linear one and proceeds with a high degree of stereocontrol throughout and an overall yield of 1.9%. The second synthesis, a block synthesis, makes use of the coupling of two trisaccharides, resulting in a shorter sequence and an overall yield of 4.4%, despite the poor selectivity in the key step. PMID- 15548006 TI - Intermolecular charge transfer and hydrogen bonding in solid furan. AB - The calculated structures of furan as a monomer, a dimer that was isolated from the crystal structure, and the full crystal structure have been thoroughly investigated by a combination of density functional theory (DFT) calculations and inelastic neutron scattering (INS) measurements. To improve our understanding of the nature and magnitude of the intermolecular interactions in the solid, the atoms in molecules (AIM) theory has been applied to the dimer and a cluster of eight monomers. After a careful topological study of the theoretical charge density and of its Laplacian, we have established the existence of C-H...pi, C H...O, and H...H interactions between adjacent molecules in solid furan. The electron distribution has also been analyzed by performing natural bond orbital (NBO) calculations for the monomer and a H-bonded dimer. When the hydrogen bond is established between two adjacent furan rings, some electron charge is transferred from the pi electronic system of one furan ring to the other molecule in the dimer. This result provides a model of the interaction between end groups of neighboring chains of polyfuran and could be applicable to other conjugated polymers where the pi system is responsible for their conducting properties. To determine how the intermolecular bonds in the solid affect the vibrational dynamics in the periodic system, INS data were analyzed by performing molecular and periodic density functional calculations. Reasonable agreement is achieved, although we note that the poorest agreement is for modes involving hydrogen atoms. PMID- 15548007 TI - Metal clips induce folding of a short unstructured peptide into an alpha-helix via turn conformations in water. Kinetic versus thermodynamic products. AB - Short peptides corresponding to two to four alpha-helical turns of proteins are not thermodynamically stable helices in water. Unstructured octapeptide Ac-His1 Ala2-Ala3-His4-His5-Glu6-Leu7-His8-NH(2) (1) reacts with two [Pd((15)NH(2)(CH(2))(2)(15)NH(2))(NO(3))(2)] in water to form a kinetically stable intermediate, [[Pden](2)[(1,4)(5,8)-peptide]](2), in which two 19-membered metallocyclic rings stabilize two peptide turns. Slow subsequent folding to a thermodynamically more stable two-turn alpha-helix drives the equilibrium to [[Pden](2)[(1,5)(4,8)-peptide]] (3), featuring two 22-membered rings. This transformation from unstructured peptide via turns to an alpha-helix suggests that metal clips might be useful probes for investigating peptide folding. PMID- 15548008 TI - Structure-based design of estrogen receptor-beta selective ligands. AB - We present the structure-based optimization of a series of estrogen receptor-beta (ERbeta) selective ligands. X-ray cocrystal structures of these ligands complexed to both ERalpha and ERbeta are described. We also discuss how molecular modeling was used to take advantage of subtle differences between the two binding cavities in order to optimize selectivity for ERbeta over ERalpha. Quantum chemical calculations are utilized to gain insight into the mechanism of selectivity enhancement. Despite only two relatively conservative residue substitutions in the ligand binding pocket, the most selective compounds have greater than 100 fold selectivity for ERbeta relative to ERalpha when measured using a competitive radioligand binding assay. PMID- 15548009 TI - Oxygen isotope effects on electron transfer to O2 probed using chemically modified flavins bound to glucose oxidase. AB - Apo-glucose oxidase has been reconstituted with flavins modified in the 7 and 8 positions and characterized with regard to the catalytic rate of O(2) reduction and oxygen-18 isotope effects on this process. Kinetic studies as a function of driving force indicate a reorganization energy for electron transfer to O(2) of lambda = 28 kcal mol(-)(1) at optimal pH, which is similar to the value obtained earlier from temperature dependencies of rates (Roth, J. P.; Klinman, J. P. Proc. Natl. Acad. Sci. U.S.A. 2003, 100, 62-67). For the various enzyme-bound flavins, competitive oxygen-18 kinetic isotope effects fall within the narrow range of 1.0266(5) to 1.0279(6), apparently because of the dominant contribution of outer sphere reorganization to the activation barrier; within the context of semiclassical and quantum mechanical electron transfer theories, the magnitude of the isotope effects reveals the importance of nuclear tunneling. PMID- 15548010 TI - Duplex formation and the onset of helicity in poly d(CG)n oligonucleotides in a solvent-free environment. AB - The gas-phase conformations of a series of cytosine/guanine DNA duplexes were examined by ion mobility and molecular dynamics methods. Deprotonated duplex ions were formed by electrospray ionization, and their collision cross sections measured in helium were compared to calculated cross sections of theoretical models generated by molecular dynamics. The 4-mer (dCGCG) and 6-mer (dCGCGCG) duplexes were found to have globular conformations. Globular and helical structures were observed for the 8-mer (dCGCGCGCG) duplex, with the globular form being the more favored conformer. For the 10-mer (dCGCGCGCGCG), 14-mer (dCGCGCGCGCGCGCG), and 18-mer (dCGCGCGCGCGCGCGCGCG) duplexes, only helical structures were observed in the ion mobility measurements. Theory predicts that the helical structures are less stable than the globular forms in the gas phase and should collapse into the globular form given enough time. However, molecular dynamics simulations at 300 K indicate the helical structures are stable in aqueous solution and will retain their conformations for a limited time in the gas phase. The presence of helical structures in the ion mobility experiments indicates that the duplexes retain "solution structures" in the gas phase on the millisecond time scale. PMID- 15548011 TI - Solvent dependence of PII conformation in model alanine peptides. AB - Alanine residues in two model peptides, the pentapeptide AcGGAGGNH(2) and the 11mer AcO(2)A(7)O(2)NH(2), have been reported to have substantial PII conformation in water. The PII structure in both peptides is sensitive to solvent. In the presence of the organic solvent TFE, the conformation of the pentamer changes from PII to internally H-bonded gamma or beta turns, while the chain with seven alanines forms alpha helix. The PII structure in the 11mer is more stable than that in the shorter peptide as the TFE concentration increases. For the pentamer, a comparison of short-chain aliphatic alcohols to water shows that the PII content decreases in the order water > methanol > ethanol > 2 propanol, linearly according to empirical scales of solvent polarity. Thus, depending on the extent of local solvation as folding progresses, the peptide backbone as modeled by alanine oligomers shifts from PII to internally H-bonded (gamma or beta turn) conformations and to alpha helix in longer segments. On the other hand, the PII content of AcO(2)A(7)O(2)NH(2) increases significantly in the presence of guanidine, as does that of oligoproline peptides, while detergent sodium dodecyl sulfate (SDS) favors alpha helix in this peptide. The shorter peptide does not show a parallel increase in PII with guanidine. PMID- 15548012 TI - [Fe2(SR)2(mu-CO)(CNMe)6]2+ and analogues: a new class of diiron dithiolates as structural models for the H(ox)Air state of the fe-only hydrogenase. AB - Low-temperature oxidation of Fe(2)(S(2)C(n)H(2n)(CNMe)(6-x)(CO)x (n = 2, 3; x = 2, 3) affords a family of mixed carbonyl-isocyanides of the type [Fe(2)(S(2)C(n)H(2n)(CO)x(CNMe)(7-x)](2+). The degree of substitution is controlled by the RNC/Fe ratio, as well as the degree of initial substitution at iron, with tricarbonyl derivatives favoring more highly carbonylated products. The structures of the monocarbonyl derivatives [Fe(2)(S(2)C(n)H(2n))(mu CO)(CNMe)(6)](PF(6))(2) (n = 2,3) established crystallographically and spectroscopically, are quite similar, with Fe---Fe distances of ca. 2.5 A, although the mu-CO is unsymmetrical in the propanedithiolate derivative. Isomeric forms of [Fe(2)(S(2)C(3)H(6))(CO)(CNMe)(6)](PF(6))(2) were characterized where the CO is bridging or terminal, the greatest structural difference being the 0.1 A elongation of the Fe---Fe distance when MeNC (vs CO) is bridging. In the dicarbonyl species, [Fe(2)(S(2)C(2)H(4))(mu-CO)(CO)(CNMe)(5)](PF(6))(2), the terminal CO ligand is situated at one of the basal sites, not trans to the Fe-- Fe vector. Oxidation of Fe(2)(S(2)C(2)H(4))(CNMe)(3)(CO)(3) under 1 atm CO gives the deep pink tricarbonyl [Fe(2)(S(2)C(2)H(4))(CO)(3)(CNMe)(4)](PF(6))(2). DFT calculations show that a bridging CO or MeNC establishes a 3-center, 2-electron bond within the two Fe(II) centers, which would otherwise be nonbonding. PMID- 15548013 TI - Macromolecular helicity induction in a cationic polyacetylene assisted by an anionic polyisocyanide with helicity memory in water: replication of macromolecular helicity. AB - We report the first example of the replication of macromolecular helicity. An optically active helical and anionic polyelectrolyte, the sodium salt of poly(4 carboxyphenyl isocyanide), was found to serve as the template for further helicity induction in a different polyelectrolyte with opposite charges in water, resulting in interpolymer helical assemblies with controlled helicity. The effects of the pH and salt concentration on the helicity induction were investigated. PMID- 15548014 TI - Solute solvent dynamics and energetics in enzyme catalysis: the S(N)2 reaction of dehalogenase as a general benchmark. AB - One of the most useful ways of describing and analyzing enzyme catalysis is the description of the enzyme as an effective solvent for the reacting substrate. Here, we illustrate this concept by considering the S(N)2 reaction of haloalkan dehalogenase (DhlA), analyze the energetics and dynamics of the solvent coordinate, and evaluate their relative catalytic effect. It is demonstrated that almost the entire catalytic effect is associated with the preorganization of the protein-solvent coordinate. It is clarified that this effect is associated with the fact that the transition state is "solvated" by the protein more than in the reference solution reaction. This effect is fundamentally different than the frequently proposed desolvation mechanism. The possible catalytic role of dynamical effects is analyzed by considering several reasonable ways of defining "dynamical contributions to catalysis". It is found that these contributions are small regardless of the definition used. It is also shown that the effect of the difference in the relaxation time of the solvent coordinate in the enzyme and solution reaction is rather trivial relative to the effect of the corresponding changes in reorganization free energy. PMID- 15548015 TI - Fluorescence and aggregation behavior of poly(amidoamine) dendrimers peripherally modified with aromatic chromophores: the effect of dendritic architectures. AB - PAMAM dendrimers of the zeroth to fifth generation (G0-5) have been peripherally modified with phenyl, naphthyl, pyrenyl, and dansyl chromophores. Their fluorescence behaviors are strongly affected by the dendritic architectures at different generations. These dendrimers modified with hydrophobic chromphores can self-organize into vesicular aggregates at the low generations G0-3 in water. The size and aggregation number of these vesicles decrease with increasing generation from G0 to G3. Critical aggregation concentration determined by fluorescence spectroscopy reveals that these aggregates can be favorably formed in the order of G3 > G2 > G1. In contrast to the vesicles made from traditional amphiphilic compounds, these dendrimer-based vesicles are very adhesive due to the H-bonding interaction and entanglement of dendritic branches located in the outer layer. A large number of multivesicle assemblies, i.e., "twins" and "quins" consisting of two and five vesicles, were clearly identifiable with transmission electron (TEM) and atomic force microscopy. For the dendrimers with peripheral pyrenyl chromophores, triangle-like vesicles were observed in water. The hydrophobic interphase thickness of the vesicular bilayer is ca. 2.0-3.2 nm determined by fluorescence resonance energy transfer methods, which agrees well with the thickness directly observed with TEM. PMID- 15548016 TI - Sterically demanding, bioxazoline-derived N-heterocyclic carbene ligands with restricted flexibility for catalysis. AB - A unique family of N-heterocyclic carbenes derived from bioxazolines (IBiox) suitable for application in transition-metal catalysis is described. The ligands are electron rich, sterically demanding, and have restricted flexibility. Their usefulness has been demonstrated in the Suzuki-Miyaura cross-coupling of sterically hindered aryl chlorides and boronic acids. For the first time, tetraortho-substituted biaryls with methyl and larger ortho-substituents have been synthesized from aryl chlorides using the Suzuki-Miyaura method. PMID- 15548017 TI - Anion binding to a ferric porphyrin complexed with per-O-methylated beta cyclodextrin in aqueous solution. AB - 5,10,15,20-Tetrakis(4-sulfonatophenyl)porphinato iron(III) (Fe(III)TPPS) forms a very stable 1:2 complex with heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (TMe beta-CD), whose iron(III) center is located at a hydrophobic cleft formed by two face-to-face TMe-beta-CD molecules. Various inorganic anions (X(-)) such as F(-), Cl(-), Br(-), I(-), N(3)(-), and SCN(-) coordinate to Fe(III)TPPS(TMe-beta-CD)(2) to form five-coordinate high-spin Fe(III)TPPS(X)(TMe-beta-CD)(2), while no coordination occurs with ClO(4)(-), H(2)PO(4)(-), NO(3)(-), and HSO(4)(-). Except for F(-), none of the anions investigated coordinate to Fe(III)TPPS in the absence of TMe-beta-CD due to extensive hydration to the anions as well as to Fe(III)TPPS. The present system shows a high selectivity toward the N(3)(-) anion. The thermodynamics suggests that Lewis basicity, hydrophilicity, and shape of an X(-) anion are the main factors to determine the stability of the Fe(III)TPPS(X)(TMe-beta-CD)(2) complex. PMID- 15548018 TI - Helically annelated and cross-conjugated oligothiophenes: asymmetric synthesis, resolution, and characterization of a carbon-sulfur [7]helicene. AB - The synthesis and characterization of a novel oligothiophene, in which the thiophene rings are annelated into a [7]helicene with cross-conjugated pi-system, are described. Such [7]helicenes may be viewed as fragments of the unprecedented carbon-sulfur (C(2)S)(n)() helix, possessing sulfur-rich molecular periphery. Racemic synthesis of [7]helicene is based upon iterative alternation of two steps: C-C bond homocouplings between the beta-positions of thiophenes and annelation between the alpha-positions of thiophenes. Asymmetric synthesis is carried out using (-)-sparteine-mediated annelation of the axially chiral bis(aryllithium) with electrophilic sulfur equivalent. Alternatively, enantiomers of the [7]helicene are obtained via resolution using menthol-based chiral siloxanes. Racemic [7]helicene and four other macrocyclic products of the annelation are characterized by X-ray crystallography. One of the solvent polymorphs of the [7]helicene possesses pi-stacked columns of opposite enantiomers and multiple short intermolecular contacts, including both homochiral and heterochiral short S...S contacts, suggesting an effective intermolecular electronic coupling in two-dimensions. The [7]helicene is configurationally stable at room temperature and racemizes at 199 degrees C with a half-time of about 11 h. Selected physicochemical studies (UV-vis absorption, CD, optical rotation, and cyclic voltammetry) of the [7]helicene are described. PMID- 15548019 TI - Lipopeptides incorporated into supported phospholipid monolayers have high specific activity at low incorporation levels. AB - The ability to present cell adhesion molecule (CAM) ligands in controlled amounts on a culture surface would greatly facilitate the control of cell growth and differentiation. Supported lipid monolayer/bilayer systems have previously been developed that allow for presentation of CAM ligands for cell interaction; however, these systems have employed peptide loadings much higher than those used in poly(ethylene glycol) (PEG)-based immobilization systems. We report the development of synthetic methods that can be used for the efficient and versatile creation of many linear and cyclic lipid-linked peptide moieties. Using RGD-based peptides for the alpha5beta1 integrin as a model system, we have demonstrated that these lipopeptides support efficient cell binding and spreading at CAM ligand loadings as low as 0.1 mol %, which is well below that previously reported for supported lipid systems. Engineered lipopeptide-based surfaces offer unique presentation options not possible with other immobilization systems, and the high activity at low loadings we have shown here may be extremely useful in presenting multiple CAM ligands for studying cell growth, differentiation, and signaling. PMID- 15548020 TI - Chelate-controlled synthesis of racemic ansa-zirconocenes. AB - The reaction of Zr[PhN(CH(2))(3)NPh]Cl(2)(THF)(2) (5) with lithium ansa-bis indenyl reagents Li(2)[XBI](Et(2)O) (XBI = (1-indenyl)(2)SiMe(2) (SBI, 7a), (2 methyl-1-indenyl)(2)SiMe(2) (MSBI, 7b), (2-methyl-4,5-benz-1-indenyl)(2)SiMe(2) (MBSBI, 7c), (2-methyl-4-phenyl-1-indenyl)(2)SiMe(2) (MPSBI, 7d), and 1,2-(1 indenyl)(2)ethane (EBI, 7e)) affords rac-(XBI)Zr[PhN(CH(2))(3)NPh] (8a-e) in high yield. The meso isomers were not detected by (1)H NMR. X-ray crystallographic studies show that the Zr[PhN(CH(2))(3)NPh] rings in 5, 8a, 8c, and (C(5)H(5))(2)Zr[PhN(CH(2))(3)NPh] (10) adopt twist conformations that position the N-Ph groups on opposite sides of the N-Zr-N plane. This conformation complements the metallocene structures of rac-8a-e but would destabilize the corresponding meso isomers. It is proposed that the Zr[PhN(CH(2))(3)NPh] ring adopts a similar twist conformation in the stereodetermining transition state for addition of the second indenyl ring in these reactions, which leads to a preference for rac products. The results of metallocene syntheses from other Zr amide precursors support this proposal. 8a-e are converted to the corresponding rac-(XBI)ZrCl(2) complexes (9a-e) by reaction with HCl. PMID- 15548021 TI - General, room-temperature method for the synthesis of isolated as well as arrays of single-crystalline ABO4-type nanorods. AB - Single-crystalline BaWO(4) and BaCrO(4) nanorods of reproducible shape and of varying sizes have been controllably prepared using a simple, room-temperature approach, based on the use of porous alumina template membranes. Aligned BaWO(4) and BaCrO(4) nanorod arrays can be obtained by dissolving the template. Our facile technique, which is analogous to biomineralization, offers a promising and generalized methodology to prepare other types of free-standing ABO(4) nanorods and their corresponding nanorod arrays. Extensive characterization of these samples has been performed using scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), energy-dispersive X-ray spectroscopy (EDS), selected area electron diffraction (SAED), Raman spectroscopy, FT-infrared spectroscopy (FT-IR), and X ray diffraction (XRD). PMID- 15548022 TI - Local measures of intermolecular free energies in solution. AB - Proton spin-lattice relaxation rate changes induced by freely diffusing oxygen in aqueous and mixed solvents are reported for representative amino acids and glucose. The local oxygen concentration at each spectrally resolved proton was deduced from the paramagnetic contribution to the relaxation rate. The measured relaxation increment is compared to that of the force-free diffusion relaxation model, and the differences are related to a free energy for the oxygen association with different portions of the solute molecules. The free energy differences are small, on the order of -800 to -2000 J/mol, but are uniformly negative for all proton positions measured on the amino acids in water and reflect the energetic benefit of weak association of hydrophobic cosolutes. For glucose, CH proton positions report negative free energies for oxygen association, the magnitude of which depends on the solvent; however, the hydroxyl positions report positive free energy differences relative to the force-free diffusion model, which is consistent with partial occupancy in the OH region by a solvent hydrogen bond. PMID- 15548023 TI - A generalized model for the molecular arrangement in the columnar mesophases of polycatenar mesogens. Crystal and molecular structure of two hexacatenar mesogens. AB - The columnar mesophases of two series of hexacatenar palladium(II) mesogens have been studied in detail by a combination of X-ray diffraction on aligned and unaligned samples and dilatometry. The results of these studies, combined with the results of two single crystal structure determinations, have allowed a model of the molecular arrangement in the columnar phases to be proposed. This model differs in detail from that generally accepted for the arrangement of polycatenar mesogens in columnar phases, and a new model is proposed which accounts for both new and existing data. PMID- 15548024 TI - Reversible surface oxidation and efficient luminescence quenching in semiconductor single-wall carbon nanotubes. AB - We have investigated reversible single-wall carbon nanotube (SWNT) oxidation by quantitative analysis of the oxide-induced absorption bleaching and luminescence quenching at low pH. These data, in combination with DFT structure calculations, suggest that the nanotube oxide is a 1,4-endoperoxide. At low pH, the endoperoxide protonates to create a hydroperoxide carbocation, introducing a hole in the SWNT valence band. Nanotube luminescence is extremely sensitive to quenching by hole-doping, while the absorption is relatively robust. PMID- 15548025 TI - Membrane localization and flexibility of a lipidated ras peptide studied by molecular dynamics simulations. AB - Lipid-modified membrane-binding proteins are essential in signal transduction events of the cell, a typical example being the GTPase ras. Recently, membrane binding of a doubly lipid-modified heptapeptide from the C-terminus of the human N-ras protein was studied by spectroscopic techniques. It was found that membrane binding is mainly due to lipid chain insertion, but it is also favored by interactions between apolar side chains and the hydrophobic region of the membrane. Here, 10 explicit solvent molecular dynamics simulations for a total time of about 150 ns are used to investigate the atomic details of the peptide membrane association. The 16:0 peptide lipid chains are more mobile than the 14:0 phospholipid chains, which is in agreement with (2)H NMR experiments. Peptide lipid and peptide-solvent interactions, backbone and side-chain distributions, as well as the effects of lipidated peptide insertion onto the structure, and dynamics of a 1,2-dimyristoylglycero-3-phosphocholine bilayer are described. The simulation results validate the structural model proposed by the analysis of spectroscopic data and highlight the main aspects of the insertion mechanism. The peptide in the membrane is rather rigid over the simulation time scale of about 10 ns, but different partially extended conformations devoid of backbone hydrogen bonds are observed in different trajectories. PMID- 15548026 TI - Development of a functionalized xenon biosensor. AB - NMR-based biosensors that utilize laser-polarized xenon offer potential advantages beyond current sensing technologies. These advantages include the capacity to simultaneously detect multiple analytes, the applicability to in vivo spectroscopy and imaging, and the possibility of "remote" amplified detection. Here, we present a detailed NMR characterization of the binding of a biotin derivatized caged-xenon sensor to avidin. Binding of "functionalized" xenon to avidin leads to a change in the chemical shift of the encapsulated xenon in addition to a broadening of the resonance, both of which serve as NMR markers of ligand-target interaction. A control experiment in which the biotin-binding site of avidin was blocked with native biotin showed no such spectral changes, confirming that only specific binding, rather than nonspecific contact, between avidin and functionalized xenon leads to the effects on the xenon NMR spectrum. The exchange rate of xenon (between solution and cage) and the xenon spin-lattice relaxation rate were not changed significantly upon binding. We describe two methods for enhancing the signal from functionalized xenon by exploiting the laser-polarized xenon magnetization reservoir. We also show that the xenon chemical shifts are distinct for xenon encapsulated in different diastereomeric cage molecules. This demonstrates the potential for tuning the encapsulated xenon chemical shift, which is a key requirement for being able to multiplex the biosensor. PMID- 15548027 TI - Preparation and characterization of pi-stacking quinodimethane oligothiophenes. Predicting semiconductor behavior and bandwidths from crystal structures and molecular orbital calculations. AB - A series of new quinodimethane-substituted terthiophene and quaterthiophene oligomers has been investigated for comparison with a previously studied quinoid oligothiophene that has demonstrated high mobilities and ambipolar transport behavior in thin-film transistor devices. Each new quinoidal thiophene derivative shows a reversible one-electron oxidation between 0.85 and 1.32 V, a quasi reversible one-electron second oxidation between 1.37 and 1.96 V, and a reversible two-electron reduction between -0.05 and -0.23 V. The solution UV-vis NIR spectrum of each compound is dominated by an intense (epsilon congruent with 100 000 M(-1) cm(-1)) low energy pi-pi transition that has a lambda(max) ranging between 648 and 790 nm. All X-ray crystal structures exhibit very planar quinoidal backbones and short intermolecular pi-stacking distances (3.335-3.492 A). Structures exhibit a single pi-stacking distance with parallel cofacial stacking (sulfur atoms of equivalent rings pointed in the same direction) or with alternating distances and antiparallel cofacial stacking (sulfur atoms of equivalent rings pointed in the opposite direction). Examples of the layered and herringbone-packing motifs are observed for both the parallel and the antiparallel cofacial stacking. Analysis of the X-ray structures and molecular orbital calculations indicates that all of these compounds have one-dimensional electronic band structures as a result of the pi-stacking. For structures with a unique pi-stacking distance, a simple geometric overlap parameter calculated from the shape of the molecule and the slip from perfect registry in the pi-stack correlates well with the transfer integrals (t) calculated using molecular orbital theory. The calculated valence (633 meV) and conduction (834 meV) bandwidths for a quinoid quaterthiophene structure are similar to those calculated for the benchmark pentacene and indicate that both hole and electron mobilities could be significant. PMID- 15548028 TI - Planar tetracoordinate carbon in extended systems. AB - A recently proposed system with a central planar tetracoordinate carbon linking two three-membered rings, C(5)(2-), lends itself to extension in one, two, and three dimensions. Our construction of potential realizations begins with an analysis of the electronic structure of C(5)(2-). Dimers such as C(10)Li(3-), C(10)Li(4), and a trimer C(15)Li(6) are then examined, and their geometries are optimized to find clues for ways the C(5)(2-) unit may polymerize in the presence of countercations. Coordination through the terminal carbons is favored in the oligomers and polymers; several electronically and structurally reasonable systems of the stoichiometry C(5)M(x) (M = Li, x = 2; M = Be, Pt, Zn, x = 1) emerge from band structure calculations and energetic considerations. PMID- 15548034 TI - Unexpected cross-coupling reaction between o-chloroaryl ketones and organomanganese reagents. AB - Alkyl- and arylmanganese reagents react with o-chloro or o-bromoaryl ketones to give the substituted ketones in high yields. The cross-coupling reaction is performed under mild conditions (-60 to +40 degrees C, 30 min to 4 h) and takes place with excellent chemoselectivity. [reaction: see text] PMID- 15548035 TI - Construction of chiral quaternary carbon centers by Pd-catalyzed asymmetric allylic substitution with P,N-1,1'-ferrocene ligands. AB - A chiral quaternary carbon center on allyl substrates was constructed by Pd catalyzed allylic alkylation reaction in good to high regio- and enantioselectivities for a wide range of substrates for the first time. [reaction: see text] PMID- 15548036 TI - Efficient conversion of O-substituted 3-hydroxy-4-imino-oxazolidin-2-ones into O substituted alpha-hydroxyamidoximes. AB - An efficient and convenient two-step synthesis of O-substituted alpha hydroxyamidoximes has been developed. The first step involves a high-yielding one pot synthesis of the so far unknown O-substituted 3-hydroxy-4-imino-oxazolidin-2 ones by reacting cyanohydrins stepwise with 1,1'-carbonyldiimidazole and O substituted hydroxylamines. The second step represents a novel, sodium methoxide mediated conversion of O-substituted 3-hydroxy-4-imino-oxazolidin-2-ones into the corresponding O-substituted alpha-hydroxyamidoximes. [reaction: see text] PMID- 15548037 TI - A new homobifunctional p-nitro phenyl ester coupling reagent for the preparation of neoglycoproteins. AB - A new linker system has been designed and applied to neoglycoprotein synthesis. Reaction of oligosaccharide omega-aminoalkyl glycosides with homobifunctional adipic acid p-nitrophenyl diesters in dry DMF gave the corresponding amide half ester in good yields and of sufficient stability to permit chromatographic purification. Subsequent conjugation with bovine serum albumin under very mild conditions afforded the corresponding neoglycoproteins with good efficiency. The method is well suited for the coupling of very small amounts (mg) of oligosaccharide and protein. [structure: see text] PMID- 15548038 TI - Synthesis of 4,4-disubstituted 2-aminocyclopentanecarboxylic acid derivatives and their incorporation into 12-helical beta-peptides. AB - An enantioselective synthetic route is reported for trans-2 aminocyclopentanecarboxylic acids (ACPC) bearing geminal side chain pairs at the 4-position. Beta-peptides containing the 4,4-disubstituted ACPC residues adopt the 12-helical conformation, as demonstrated by 2D NMR analysis in aqueous solution. These 4,4-disubstituted ACPC residues display functional groups, including acidic and hydrogen bond donating groups, in a geometrically defined fashion, which should be useful for the design of beta-peptides for specific applications. [structure: see text] PMID- 15548039 TI - A four-component one-pot synthesis of alpha-Gal pentasaccharide. AB - A four-component one-pot sequential synthesis of alpha-Gal pentasaccharide 2 with minimal protecting group manipulations in a very short period of time is described in this paper. [structure: see text] PMID- 15548040 TI - Palladium-catalyzed borylation of phenyl bromides and application in one-pot Suzuki-Miyaura biphenyl synthesis. AB - The coupling reaction of pinacolborane with various aryl bromides in the presence of a catalytic amount of Pd(OAc)(2) together with DPEphos as ligand and Et(3)N as base provided arylboronates. High yields were obtained in the case of electron donor substituted aryl bromides. The direct preparation of arylboronates allowed the one-pot, two-step synthesis of unsymmetrical biaryls in high yields. [reaction: see text] PMID- 15548041 TI - CamTHP*OH: a camphor-derived delta-lactol auxiliary for the effective desymmetrization of attached glycinamide residues. Asymmetric synthesis of alpha amino carbonyl compounds. AB - Stereoselective allylation of camphor and subsequent terminal hydroformylation affords a new delta-lactol auxiliary (camTHP*OH) on multigram scale. Stereoselective condensation with glycine dimethylamide and Cbz protection affords a camTHP*-desymmetrized glycinamide building block which undergoes efficient and highly diastereoselective metal enolate alkylation reactions. Acid mediated deprotection affords the N-Cbz-protected alpha-amino amide products which may be converted directly to alpha-amino ketones on treatment with Grignard or organolithium reagents without loss of stereochemical integrity. [reaction: see text] PMID- 15548042 TI - Highly stereoselective Michael addition reactions of CamTHP*-desymmetrized glycinamide for the synthesis of functionally dense amino acid derivatives. AB - The camphor-derived tetrahydropyran (camTHP*)-desymmetrized glycinamide 1 undergoes efficient and highly diastereoselective lithium enolate Michael additions to nitro olefins, alpha,beta-unsaturated ketones, esters, and lactones. Straightforward manipulation of these products affords 3-substituted pyroglutamides and beta-aryl-alpha,gamma-diamino acid derivatives, highlighting the ease of synthesis of enantiomerically enriched, functionally dense molecules using this novel building block. [reaction: see text] PMID- 15548043 TI - Unusually stable molecular capsule formation of a tetraphenyleneurea cavitand. AB - An unusually stable molecular capsule was formed by heating phenyleneurea-spanned resorcinarene cavitand with 4-methyl-N-p-tolylbenzamide. The molecular capsule behaved as a discrete molecular entity showing a cylindrical D(4d) structure and showed no guest exchange in toluene-d(8) even at 100 degrees C. [structure: see text] PMID- 15548044 TI - Eta2-dba complexes of Pd(0): the substituent effect in Suzuki-Miyaura coupling. AB - The influence of aryl substituents in dibenzylidene acetone (dba) ligands, for Pd(0) complexes, has been evaluated for Suzuki-Miyaura cross-coupling reactions. Electron-withdrawing substituents such as NO(2) or CF(3) deactivate the catalyst species whereas strongly donating substituents such as OMe increase catalytic activity over that of unsubstituted dba ligands. [reaction: see text] PMID- 15548045 TI - Asymmetric synthesis of deoxypolypropionate units via stereoselective hydrogenation of optically active cycloheptatriene. AB - Optically active polypropionate units were synthesized in 9-11 steps from 3,5 dimethylphenol. The sequence consists of the Buchner reaction controlled by a chiral 2,4-pentanediol tether and diastereoselective hydrogenation over Raney nickel. [reaction: see text] PMID- 15548046 TI - Synthesis of type II beta-turn surrogate dipeptides based on syn-alpha-amino alpha,beta-dialkyl-beta-lactams. AB - The achiral bis(trimethylsilyl)methyl group acts as an efficient stereochemical determinant of the alpha-alkylation reaction in beta-branched alpha phenyloxazolidinyl- or alpha-diphenyloxazolidinyl-beta-lactams and provides the first stereocontrolled access to syn-alpha-amino-alpha,beta-dialkyl(aryl)-beta lactams. These products are readily transformed into type II beta-turn mimetic surrogates 2B. [reaction: see text] PMID- 15548047 TI - Beta-1,3-glucan (schizophyllan) can act as a one-dimensional host for creation of novel poly(aniline) nanofiber structures. AB - We here demonstrate the creation of novel poly(aniline) (PANI) nanofiber structures by a polymer wrapping method using schizophyllan (SPG). Mannose modified SPG can also wrap PANIs to give nanofibers having a lectin affinity. This interaction is applicable to designing novel PANI/protein composites. The results establish that SPG can act as a novel "host" to assemble PANIs into one dimensional superstructures. [reaction: see text] PMID- 15548048 TI - Ring-current models from the differential Biot-Savart law. AB - The differential Biot-Savart law provides simple models for the pi ring currents induced in diatropic and paratropic planar conjugated molecules by a perpendicular magnetic field. The model predictions are confirmed by ab initio maps of nuclear magnetic shielding density. The effects on the protons and on the ring carbon atoms from the closest and furthest segments of the current loop are easily interpreted. [structure: see text] PMID- 15548049 TI - Synthesis of novel 6,11-O-bridged bicyclic ketolides via a palladium-catalyzed bis-allylation. AB - A bridging chemistry process was developed to form an ether bridge between 6-O and 11-O of erythromycin A via a tandem or stepwise palladium-catalyzed bis-pi allylation. By applying this bridging process, new 6,11-O-bridged bicyclic ketolides (BBKs) were synthesized. These BBKs showed good antibacterial activities against the macrolide-susceptible strains as well as mef-resistant strains and served as a good core for further modifications to study the structure-activity relationship (SAR) and to overcome bacterial resistance. [reaction: see text] PMID- 15548050 TI - Design, synthesis, and evaluation of a radicicol and geldanamycin chimera, radamide. AB - A chimera composed of the natural products radicicol and geldanamycin has been prepared through an amide linkage connecting the resorcinol moiety of radicicol to the quinone ring of geldanamycin. The inhibitory activity of these compounds was determined by their ability to inhibit Hsp90's inherent ATPase activity along with degradation of the Hsp90 client protein, HER-2 in MCF-7 breast cancer cells. [reaction: see text] PMID- 15548051 TI - A new motif in the formation of peptide nanotubes: the crystallographic signature. AB - Terminally protected acyclic tripeptides Boc-Tyr(1)-Val(2)-Tyr(3)-OMe 1 and Boc Tyr(1)-Ile(2)-Tyr(3)-OMe 2 self-assemble into nanotubes in crystals through various noncovalent interactions with an average internal diameter of 5 A (0.5 nm), and the tubular ensemble is developed through the hydrogen-bonded side chains of tyrosine residues. The inside of the hollow nanotubular structures is hydrophilic; however, no solvent molecules have been crystallographically detected. [structure: see text] PMID- 15548052 TI - High stereoselective preparation of O-protected 2-trifluoromethyl 3-bromoallylic alcohols from 1,1-dibromo-1-alkenes. AB - A highly stereoselective lithium-bromine exchange reaction of 2-trifluoromethyl 3,3-dibromoallylic alcohols is described. (E)- and (Z)-2-trifluoro-3-bromoallylic alcohols were obtained in THF and hexane, respectively. The lithium carbenoid intermediate was stable even at -40 degrees C and could be trapped by various electrophiles to afford functionalized 2-trifluoromethyl-3-bromoallylic alcohols. [reaction: see text] PMID- 15548054 TI - In- or in(I)-employed diastereoselective Reformatsky-type reactions with ketones: 1H NMR investigations on the active species. AB - An efficient In- or In(I)-based stereoselective C-C bond formation is reported; the diastereoselective Reformatsky-type reactions of ketones. The predominant formations of anti isomers, confirmed by the X-ray structure analyses of ester derivatives of respective alcohols 9a(1)() and 13a(1)(), conclusively revealed the stereochemistry of the reaction path. (1)H NMR investigations revealed the formation of two types of alpha-metalated transient species from alpha-halo esters with In or In(I) halides. [reaction: see text] PMID- 15548053 TI - Site-selective DNA photocleavage involving unusual photoinitiated tautomerization of chiral tridentate vanadyl(V) complexes derived from N-salicylidene alpha-amino acids. AB - The titled vanadyl(V) complexes serve as efficient reagents for cleaving supercoiled plasmid DNA by photoinitiation. Complex 3d, derived from 2-hydroxy-1 naphthaldehyde and l-phenylalanine, exhibits a unique wedge feature, inducing a site-selective photocleavage at the C22-T23 of the bulge backbone for a HIV-27 DNA system at 0.1-5 muM. Transient absorption experiments for 3d indicate the involvement of LMCT with concomitant tautomerization, leading to an o-quinone methide V-bound hydroxyl species responsible for the cleavage profiles. [structure: see text] PMID- 15548055 TI - Stereoselective N-glycosylation by Staudinger ligation. AB - Stereoselective methods for the chemical synthesis of beta-N-glycosyl amides are needed to generate glycopeptides and glycoproteins. Here, we report that the Staudinger ligation can be used to form glycosylated asparagine derivatives. The reaction proceeds with high stereoselectivity, and a variety of glycosyl azides can function as substrates. Our results provide precedence for the use of this powerful amide-bond-forming reaction for N-glycopeptide synthesis. [reaction: see text] PMID- 15548056 TI - Facile synthesis of @-tide beta-strand peptidomimetics: improved assembly in solution and on solid phase. AB - The synthesis of @-tide beta-strand peptidomimetics has been improved such that oligomers now can be obtained from solution- and solid-phase synthesis protocols approaching the efficiency and flexibility of peptide chemistry. These methods enable the synthesis of @-tide oligomers with a variety of amino acids and with lengths up to 13 units. [reaction: see text] PMID- 15548057 TI - Alkene substituents for selective activation of endo-regioselective polyepoxide oxacyclizations. AB - The presence of an alkenyl substituent on the terminal epoxide of a polyepoxide substrate enhances the yield of all-endo-regioselective tandem oxacyclization to trans-syn-trans-fused polycyclic ethers. For a substrate in which the epoxide and alkene functional groups are separated by two methylene substituents, a novel bromonium ion-induced endo-regioselective cyclization to bromooxepane is also described. [reaction: see text] PMID- 15548058 TI - Synthetic studies on the taxane skeleton: construction of eight-membered carbocyclic rings by the intramolecular B-alkyl Suzuki-Miyaura cross-coupling reaction. AB - Construction of eight-membered carbocyclic rings via the intramolecular B-alkyl Suzuki-Miyaura cross-coupling reaction has been studied. This protocol proved its potency through the formation of the eight-membered ring possessing a quaternary carbon on its ring in high yield, affording promise of a new access to the eight membered ring of Taxol. [reaction: see text] PMID- 15548059 TI - Formal [3 + 2] and [3 + 3] additions of acceptor-substituted cyclopropylmethylsilanes to allenylsilanes. AB - 1,3-Dipolar synthons formed from vicinal TBDPS-substituted cyclopropyl alkyl/phenyl ketones on treatment with Lewis acids such as TiCl(4) and Et(2)AlCl reacted with allenylsilanes to furnish [3 + 2] and [3 + 3] adducts with high regio- and stereocontrol. [reaction: see text] PMID- 15548060 TI - Synthesis of enantiopure imidazolines through a Ritter reaction of 2-(1 aminoalkyl)aziridines with nitriles. AB - The Ritter reaction of enantiopure 2-(1-aminoalkyl)aziridines 1 with different nitriles afford enantiopure tetrasubstituted imidazolines 2. The opening of the aziridine ring takes place with total regio- and stereoselectivity. A mechanism to explain the described addition reaction is proposed. [reaction: see text] PMID- 15548061 TI - Sulfonimidamides: efficient chiral iminoiodane precursors for diastereoselective copper-catalyzed aziridination of olefins. AB - N-(p-Toluenesulfonyl)-p-toluenesulfonimidamide reacts with iodosylbenzene to afford in situ a chiral iminoiodane. The latter gives, in the presence of a copper(I) catalyst, a nitrene that is very efficiently transferred under stoichiometric conditions to a variety of alkenes with diastereoselectivities up to 60%. [reaction: see text] PMID- 15548062 TI - Is the tert-butyl group bulky enough to end-cap a pseudorotaxane with a 24-crown 8-ether wheel? AB - Although rotaxane chemists have long believed that the tert-butyl group is bulkier than the cavity of dibenzo-24-crown-8-ether (DB24C8), it is essentially smaller than the cavity of DB24C8. The tert-butyl (or 4-tert-butylphenyl) group can actually function as an end-cap of DB24C8-based rotaxanes when the intercomponent interaction is effectively operative. When such attractive interaction is removed, deslippage occurs. [structure: see text] PMID- 15548063 TI - Lewis acid catalysis in heterolysis reactions of glycol ether radicals mimicking diol dehydratase-catalyzed reactions. AB - Zinc bromide-catalyzed heterolysis reactions of glycol ether radicals were studied by laser flash photolysis methods, which gave the binding constants and catalytic rate constants for fragmentation. The Lewis acid-catalyzed heterolysis reactions mimic a putative reaction pathway in diol dehydratase-catalyzed reactions and are potentially useful polar processes for incorporation into conventional radical chain reaction sequences. [reaction: see text] PMID- 15548064 TI - A novel strategy for oligosaccharide synthesis via temporarily deactivated S thiazolyl glycosides as glycosyl acceptors. AB - A new glycosylation strategy that allows chemoselective activation of the S thiazolyl (STaz) moiety of a glycosyl donor over the temporarily deactivated glycosyl acceptor, bearing the same anomeric group, has been developed. This deactivation is achieved by engaging of the STaz moiety of the glycosyl acceptor into a stable palladium(II) complex. Therefore, obtained disaccharides are then released from the complex by simple ligand exchange. [reaction: see text] PMID- 15548065 TI - A new class of near-infrared electrochromic oxamide-based dinuclear ruthenium complexes. AB - We report the synthesis of a new class of symmetric and unsymmetric oxamide-based dinuclear ruthenium complexes. These complexes were characterized by NMR, ESI-MS, and electrochemical methods. Spectroelectrochemical analysis of the complexes showed broad absorptions in the NIR region for the mixed-valence state of the complexes. The introduction of a chiral group into the bridging ligand produced an optically active complex that was studied using circular dichroism. [structure: see text] PMID- 15548066 TI - Asymmetric synthesis of cis-5-substituted pyrrolidine 2-phosphonates using metal carbenoid NH insertion and delta-amino beta-ketophosphonates. AB - Cis-5-substituted pyrrolidine phosphonates, proline surrogates, are prepared by a highly stereoselective intramolecular metal carbenoid N-H insertion from a sulfinimine-derived delta-amino alpha-diazo beta-ketophosphonate. [reaction: see text] PMID- 15548067 TI - Rearrangements encountered in the attempted syntheses of pyridoazepinone carboxylic acids. AB - Attempts to synthesize pyridoazepinone carboxylic acids such as 33 by standard methodologies resulted exclusively in unusual and unexpected rearrangement products. Seven-membered ring formation was attempted by ring expansion of a six membered ring and by aldol ring closure. In each case, the major product resulted from rearrangement of the starting material without detection of the desired product. Ultimately, an isomeric pyridoazepinone ethyl ester was prepared; however, attempted saponification resulted in another unusual rearrangement. [reaction: see text] PMID- 15548068 TI - A novel carbon skeletal trichothecane, tenuipesine A, isolated from an entomopathogenic fungus, Paecilomyces tenuipes. AB - Tenuipesine A (1), a novel trichothecane with an unprecedented carbon-migrated skeleton that embodies of a cyclopropane ring, was isolated from cultivated fruiting bodies of Paecilomyces tenuipes (Isaria japonica), a popular entomopathogenic fungi employed in folk medicine and health foods in China, Korea, and Japan. The structure was determined on the basis of two-dimensional NMR data. Its stereochemistry was elucidated by spectroscopic data and the chemical transformation of the coexisting trichothecene, 4beta-acetoxy-12,13 epoxytrichothec-9-ene-3alpha,15-diol (2). [structure: see text] PMID- 15548069 TI - Stereoselective total synthesis of ent-EI-1941-2 and Epi-ent-EI-1941-2. AB - The first asymmetric total syntheses of ent-EI-1941-2 and epi-ent-EI-1941-2 have been accomplished, starting from a chiral epoxy iodoquinone 6, a key intermediate in our total syntheses of epoxyquinols A and B. A key step in the preparation of ent-EI-1941-2 is an intramolecular carboxypalladation via a 6-endo cyclization mode, followed by beta-hydride elimination, while carboxymercuration is a key step in the synthesis of epi-ent-EI-1941-2. [reaction: see text] PMID- 15548070 TI - A concise asymmetric synthesis of the ADE fragment of nakadomarin A. AB - The ADE fragment of nakadomarin A has been synthesized in nine linear steps from commercial material. The key transformation is an asymmetric azomethine ylide [1,3]-dipolar cycloaddition to establish the AD-spirocyclic system containing three of the four stereocenters of the natural product. [reaction: see text] PMID- 15548071 TI - Ruthenium-catalyzed [2 + 2] cycloadditions between bicyclic alkenes and alkynyl halides. AB - Ru-catalyzed [2 + 2] cycloadditions between norbornadiene and alkynyl halides were found to occur in moderate to good yields (32-89%). The presence of the halide moiety greatly enhances the reactivity of the alkyne component in the cycloaddition and can be transformed into a variety of products that are difficult or impossible to obtain via direct cycloaddition. [reaction: see text] PMID- 15548072 TI - Alpha-arylsulfanyl-alpha-fluoro carbenoids: their novel chemistry and synthetic applications. AB - The bromine-magnesium exchange reactions of arylthiobromodifluoromethanes with Grignard reagents have been studied. Upon trapping with electrophiles, alkyl aryl sulfides and ketenedithioacetals are obtained. The reaction is proposed to occur via novel alpha-arylsulfanyl-alpha-fluoro carbenoids. The first examples of arylthiomethane multipole synthons are also reported. [reaction: see text] PMID- 15548073 TI - Method for the rapid synthesis of highly functionalized 2-hydroxy-1-naphthoates. Syntheses of the naphthoic acid components of neocarzinostatin chromophore and N1999A2. AB - We describe a four-step sequence for the synthesis of complex 2-hydroxy-1 naphthoic acids involving Z-selective olefination of benzaldehyde derivatives with a novel dioxolenone-containing phenyl phosphonate reagent, followed by dioxolenone cleavage with alkaline trifluoroethanol and oxidative cyclization (Mn(OAc)(3)) of the resultant trifluoroethyl beta-keto esters. [reaction: see text] PMID- 15548074 TI - Radical alkenylation of alpha-halo carbonyl compounds with alkenylindiums. AB - Alkenylation reaction of alpha-halo carbonyl compounds with alkenylindiums proceeded via a radical process in the presence of triethylborane. Unactivated alkene moieties as well as a styryl group could be introduced by this method. The geometry of the carbon-carbon double bonds of the alkenylindiums was retained. Preparation of an alkenylindium via a hydroindation of 1-alkyne followed by radical alkenylation established an efficient one-pot strategy. [reaction: see text] PMID- 15548075 TI - Modular synthesis of fluorous trialkylphosphines. AB - A modular synthetic protocol was developed for the preparation of fluorous trialkylphosphines with a different number of methylene spacers and various lengths of the fluorous ponytails P[(CH(2))(A)R(FX)][(CH(2))(B)R(FY)][(CH(2))(C)R(FZ)] (A, B, C = 3 and 4; X, Y, Z = 4, 6, and 8). [reaction: see text] PMID- 15548076 TI - 1,8-bis(diphenylmethylium)naphthalenediyl dication as an organic oxidant: synthesis of benzidines via self-coupling of N,N-dialkylanilines. AB - 1,8-Bis(diphenylmethylium)naphthalenediyl dications promoted oxidative coupling of various N,N-dialkylanilines to afford the corresponding para-coupled bisanilines, benzidines, in good to excellent yield. The dications were readily prepared from a cyclic ether precursor, derived from 1,8-dibromonaphthalene, via deoxygenation with a silylating reagent in 1,1,1,3,3,3-hexafluoro-2-propanol under anhydrous conditions. [reaction: see text] PMID- 15548077 TI - Synthesis of rigid multivalent scaffolds based on adamantane. AB - An efficient route to novel 1,3,5,7-tetrasubstituted derivatives of adamantane is described. This route starts from adamantane and gives the tetrafunctionalized derivative 9 in eight steps with an overall yield of 23%. These tetrahedrally shaped molecules possess three identical arms terminated by an activated carboxylic acid derivative and a protected amino function in the 1-position. We propose these tetravalent cage compounds such as 9 as scaffolds for the assembly of ligand/marker conjugates for studies of multivalent ligand receptor interactions. [reaction: see text] PMID- 15548078 TI - First asymmetric synthesis of orthoquinone monoketal enantiomers via anodic oxidation. AB - An asymmetric synthesis of orthoquinone monoketals was accomplished using anodic oxidation to convert aryl methyl ethers bearing a chiral ethanol unit into orthoquinone bisketals, followed by monohydrolysis of their dimethyl ketal unit. All four possible stereoisomers were generated in a diastereoselective manner by varying the attachment point of the chiral pro-ketal alcoholic auxiliary to the starting arene. A preliminary screening of subsequent nucleophilic addition reactions confirmed the potential utility of these synthons in asymmetric synthesis. [reaction: see text] PMID- 15548079 TI - Convenient catalytic, enantioselective conjugate reduction of nitroalkenes using CuF2. AB - We document the use of a new catalyst system for the enantioselective conjugate reduction of nitroalkenes utilizing commercially available CuF(2) and the bis phosphine JOSIPHOS as a ligand. This new protocol not only facilitates ready access to a variety of optically active nitroalkanes, but the results provide also new insight into copper-catalyzed reactions. [reaction: see text] PMID- 15548080 TI - Polymer-supported Mukaiyama reagent: a useful coupling reagent for the synthesis of esters and amides. AB - Polymer-supported N-alkyl-2-chloro pyridinium triflate was synthesized in one step from Wang resin. This reagent proved to be a very effective coupling reagent for the synthesis of esters or amides from carboxylic acids and alcohols or amines (primary and secondary). [reaction: see text] PMID- 15548081 TI - Reducing activity of polyphenols with stable radicals of the TTM series. Electron transfer versus H-abstraction reactions in flavan-3-ols. AB - A new method to test the antioxidant activity of polyphenols by electron transfer reactions to a stable organic free radical, tris(2,4,6-trichloro-3,5 dinitrophenyl)methyl radical (HNTTM), is reported. Therefore, the activity of the natural flavanols, (-)-epicatechin, and two synthetic derivatives, 4beta-(S cysteinyl)epicatechin and 4beta-(2-aminoethylthio)epicatechin, can be differentiated by their capacity to transfer hydrogen atoms to 2,2-diphenyl-1 picrylhydrazyl radical (DPPH) and to transfer electrons to HNTTM. [structure: see text] PMID- 15548082 TI - A general palladium-catalyzed coupling of aryl bromides/triflates and thiols. AB - We have developed an efficient palladium-catalyzed carbon-sulfur bond formation reaction of aryl bromides, triflates, and activated aryl chloride. Using this protocol, we have shown tolerance to a wide variety of aryl thiols and alkyl thiols that can also be used as sulfide equivalents. [reaction: see text] PMID- 15548083 TI - Buttressing effects on haloarene deprotonation: a merely kinetic or also thermodynamic phenomenon? AB - (2,6-Dichlorophenyl)- and (2,6-dibromophenyl)trialkylsilanes undergo hydrogen/metal interconversion preferentially at the 4- rather than 3-position. However, the organometallic species generated by such a "meta metalation" are thermodynamically less stable (i.e., more basic) than those that would result from an ordinary "ortho metalation". This was demonstrated by equilibration experiments based on permutational halogen/metal interconversion. A new buttressing effect can explain the unprecedented regioselectivity. It is supported by X-ray structures that reveal marked deformations of the benzene ring in halophenylsilanes. [structure: see text] PMID- 15548084 TI - Stereoselective synthesis of (+/-)-rocaglaol analogues. AB - An intramolecular hydroxy epoxide opening was used to access the cyclopenta[b]benzofuran ring system of the natural product rocaglaol (2). Our route allowed the stereocontrolled preparation of the rocaglaol derivative (+/-) (1S*,3S*,3aR*,8bS*)-3b. The synthesis of the (+/-)-(3R*)-epimer of 3b was also achieved. Our strategy is well-suited for the production of analogues with variation of the western ring. [reaction: see text] PMID- 15548085 TI - Selective fluorescence sensing of Li+ in an aqueous environment by a ferrocene anthracene-linked dyad. AB - A new 1-(9-anthryl)-4-ferrocenyl-2-aza-1,3-butadiene showing selective fluorescence enhancement upon binding to lithium cations over other alkaline cations in aqueous environment has been conveniently synthesized and characterized. [structure: see text] PMID- 15548086 TI - A fluorogenic 1,3-dipolar cycloaddition reaction of 3-azidocoumarins and acetylenes. AB - Copper(I)-catalyzed 1,3-dipolar cycloaddition reaction of nonfluorescent 3 azidocoumarins and terminal alkynes afforded intense fluorescent 1,2,3-triazole products. The mild condition of this reaction allowed us to construct a large library of pure fluorescent coumarin dyes. Since both azide and alkyne are quite inert to biological systems, this reaction has potential in bioconjugation and bioimaging applications. [reaction: see text] PMID- 15548087 TI - A very reliable method for determination of absolute configuration of chiral secondary alcohols by 1H NMR spectroscopy. AB - Surprisingly stable synperiplanar conformers of CFTA esters have led us to develop a new and very reliable method for assigning absolute configurations of even secondary alcohols having minimal structural differences, such as chiral benzhydrols and alpha-monodeuterated benzyl alcohols. PMID- 15548088 TI - Novel cyclooctatetraene radical cation planarized by full annelation with bicyclo[2.1.1]hexene units. AB - A novel cyclooctatetraene (COT) radical cation fully annelated with bicyclo[2.1.1]hexene units was prepared as SbCl(6)(-) salt, and planarity of the octagonal ring was clarified by ESR and theoretical calculations. Its longest wavelength absorption (630 nm) is blue-shifted from that (745 nm) of COT radical cation annelated with bicyclo[2.2.2]octene units due to the widening of the HOMO SOMO gap accompanying the flattening of the COT ring. [structure: see text] PMID- 15548089 TI - Chemical mediators: the remarkable structure and host-selectivity of depsilairdin, a sesquiterpenic depsipeptide containing a new amino acid. AB - The chemical structure determination of depsilairdin, a highly selective phytotoxin produced by the plant pathogenic fungus Leptosphaeriamaculans/Phoma lingam, is described. The elucidation of the unusual chemical structure used a combination of NMR spectral data and X-ray crystallography. The absolute configuration was established using chemical degradation and synthesis of (3S,6R) 3,6-diisopropyl-2,5-morpholinedione and its (3R,6S) and (3R,6R) stereoisomers. Similar to the fungal pathogen, depsilairdin caused strong lesions only on brown mustard leaves but not on related species. [structure: see text] PMID- 15548090 TI - Synthesis of primary amines by the electrophilic amination of Grignard reagents with 1,3-dioxolan-2-one O-sulfonyloxime. AB - Primary amines are prepared by the electrophilic amination of Grignard reagents with 4,4,5,5-tetramethyl-1,3-dioxolan-2-one O-phenylsulfonyloxime and the acidic hydrolysis of the resulting imines. [reaction: see text] PMID- 15548091 TI - Oxidative cross-coupling of acrylates with vinyl carboxylates catalyzed by a Pd(OAc)2/HPMoV/O2 system. AB - Oxidative cross-coupling of acrylates with vinyl carboxylates was first successfully achieved by the use of a Pd(OAc)(2)/HPMoV/O(2) system in fair to good yields. For instance, the reaction of n-butyl acrylate with vinyl acetate in the presence of catalytic amounts of Pd(OAc)(2) and H(4)PMo(11)VO(40).nH(2)O under O(2) in acetic acid at 70 degrees C for 12 h afforded the corresponding cross-coupling product, n-butyl 5-(acetoxy)-2,4-pentadienoate, in 70% yield. PMID- 15548092 TI - N-h insertion reactions of Boc-amino acid amides: solution- and solid-phase synthesis of pyrazinones and pyrazines. AB - A series of alpha-diazo-beta-ketoesters were reacted with Boc amino acid amides in the presence of rhodium octanoate catalyst. The resulting N-H insertion products were treated with acid, providing the 1,4-azine intermediates, which were oxidized by air to form the corresponding pyrazine-6-one products. The pyrazine-6-ones were further derivatized by N-alkylation or by conversion to the arylpyrazines using sequential bromination and Suzuki coupling reactions. [reaction: see text] PMID- 15548093 TI - Utility of the iridium complex of the pybox ligand in regio- and enantioselective allylic substitution. AB - The viability of the iridium complex of pybox as chiral catalyst in allylic substitutions and the enantioselective synthesis of branched products was studied. Among several chiral ligands evaluated, the iridium complex of pybox having a phenyl group catalyzed the reaction with high activity to form the branched amines with good enantioselectivities when hydroxylamine, amine, and aniline were employed as a nucleophile. The allylic substitution with oximes proceeded smoothly to give the branched oxime ethers with good enantioselectivities. [reaction: see text] PMID- 15548095 TI - Letter from the editor: Mental health issues for children and adolescents. PMID- 15548096 TI - Raising the bar of pediatric patient safety: report from the Institute of Medicine. PMID- 15548097 TI - Training in community pediatrics: a national survey of program directors. AB - OBJECTIVE: To describe the spectrum of residency training in community-based settings, assess the extent of resident education on community pediatrics topics, and determine whether educational activities vary by program size or availability of primary care tracks. METHODS: Survey of US pediatric residency program directors from May-September 2002. A 10-item self-administered questionnaire assessed the programs' extent of resident involvement in 15 selected community based settings and inclusion of didactic or practical education regarding 13 community health topics. RESULTS: Of 168 programs surveyed (81% response rate), 40% were small (< or =30 residents), 35% were medium (31-50 residents), 25% were large (>50 residents), and 15% had primary care tracks. Frequently required community-based settings included schools (69%), child protection teams (62%), day care centers (57%), and home visiting (48%). Of 15 community-based settings, 28% required involvement in fewer than 4, 41% required involvement in 4-6, and 31% required involvement in 7 or more. More than two-thirds offered didactic teaching and practical experience on issues related to managed care, cultural competency, and the mental health and social service systems. There were no differences in the number of required community-based settings by program size or presence of primary care tracks. CONCLUSIONS: Most pediatric residency programs require exposure to community-based settings and provide education on various community health topics. Ongoing challenges include continued implementation amid work duty hour limitations, best practice models for practical implementation of community-based experience into residency training, and the impact of such training on future involvement in the community and physician practice. PMID- 15548098 TI - Pediatricians' recommendations for complementary and alternative medical (CAM) therapies. AB - OBJECTIVES: Increasing numbers of children use complementary and alternative medicine (CAM). The purpose of this project was to assess pediatricians' attitudes and recommendations regarding CAM. METHODS: This was a national survey sent to a random sample of active members of the American Academy of Pediatrics (AAP). The survey was 8 pages long and included questions on demographic and practice characteristics, attitudes about CAM, and 3 clinical vignettes asking what therapies physicians recommended in these situations. RESULTS: The 745 eligible respondents were demographically similar to the national AAP membership. Most (87%) had been asked about CAM by a patient/parent in the 3 months prior to the survey and 34% said they or an immediate family member had used CAM in the past year. Most (66%) believed that CAM therapies could enhance recovery or relieve symptoms, but even more were concerned about possible side effects (75%) or that CAM use might delay mainstream care (74%). Only 20% routinely asked patients/parents about their use of herbs, and fewer asked about other CAM therapies. Fewer than 5% felt very knowledgeable about individual CAM therapies. For the clinical vignettes, pediatricians were far more likely to recommend medications than any CAM therapy. Over 80% of pediatricians desired additional information on CAM; the highest priorities were information about herbs, dietary supplements, nutritional therapies, and therapeutic exercise. CONCLUSIONS: Pediatricians recognize that many patients are interested in using CAM therapies, but do not feel comfortable discussing or recommending CAM therapies. Pediatricians are very interested in learning more about CAM. PMID- 15548099 TI - Communicating with pediatricians about complementary/alternative medicine: perspectives from parents of children with down syndrome. AB - OBJECTIVES: Barriers to communication about complementary/alternative medicine (CAM) between parents and pediatricians are frequently documented, yet the scope of these barriers remains poorly understood. Such barriers are especially troubling when they involve children with special health needs, among whom CAM use is especially common. This pilot study of parents of children with Down syndrome (DS) used qualitative methods to explore parents' perceptions of the extent and quality of communication about CAM with pediatricians, to elicit parents' recommendations for improvement, and to formulate new research questions. DESIGN: Semistructured interviews were conducted with parents from 30 families with children with DS. Data were audiotaped and analyzed with assistance from qualitative data analysis software. RESULTS: Parents described how they advocated vigorously with their pediatricians about biomedical concerns such as the American Academy of Pediatrics healthcare guidelines for DS, but often avoided discussion of nonbiomedical concerns such as CAM. Many parents looked to pediatricians to initiate conversations about CAM. DISCUSSION: Even parents who assertively advocate for biomedical concerns in their children's health care may be unlikely to disclose and discuss CAM use with their pediatricians. Attending to parents' experiences helps to illuminate the nature and scope of current communication barriers and poses new research questions for assessing and improving parent-physician collaboration about health-related issues that may be prioritized differently by parents and pediatricians. PMID- 15548100 TI - Incidence and description of scooter-related injuries among children. AB - OBJECTIVE: To describe trends in scooter-related injuries among US youth and compare scooter injuries to those related to in-line skates and skateboards. DESIGN: Retrospective review of data for children 1-19 years old from the National Electronic Injury Surveillance System of the US Consumer Product Safety Commission for 1997-2002. RESULTS: There were an estimated 190,878 scooter related injuries (95% confidence interval: 145,984-235,773) among children treated in US emergency departments; 90% were in those 15 years old or younger. There was a marked increase in scooter-related injuries in 2000, injuries peaked in 2001, and declined. In 2002, the number of scooter-related injuries was similar to in-line skates and lower than skateboard-related injuries. Scooter- and in-line skate-associated injuries primarily involved children 5-12 years old: rates of scooter-related injuries were higher than rates of in-line skate associated injuries among those 1-9 years old. Skateboard-related injuries more often involved teens. Forearm fractures accounted for 56% of fractures related to scooters (vs 74% in-line skates and 49% skateboards, chi-square, P < .01). Five percent of children injured using scooters had a closed head injury or skull fracture, similar to in-line skates and skateboards. CONCLUSIONS: The annual number of injuries related to scooters, which peaked in 2001, is now similar to the number of injuries related to in-line skates. Injuries related to scooters primarily involve children 5-12 years old, and forearm fractures are common. These data suggest helmets should be used, and protective equipment should be developed to reduce forearm fractures. PMID- 15548101 TI - Satisfaction with provider communication among Spanish-speaking Medicaid enrollees. AB - OBJECTIVE: To determine if differences between English- and Spanish-speaking parents in ratings of their children's health care can be explained by need for interpretive services. METHODS: Using the Consumer Assessment of Health Plans Survey-Child-Survey (CAHPS), reports about provider communication were compared among 3 groups of parents enrolled in a Medicaid managed care health plan: 1) English speakers, 2) Spanish speakers with no self-reported need for interpretive services, and 3) Spanish speakers with self-reported need for interpretive services. Parents were asked to report how well their providers 1) listened carefully to what was being said, 2) explained things in a way that could be understood, 3) respected their comments and concerns, and 4) spent enough time during medical encounters. Multivariate logistic regression was used to compare the ratings of each of the 3 groups while controlling for child's gender, parent's gender, parent's educational attainment, child's health status, and survey year. RESULTS: Spanish-speaking parents in need of interpretive services were less likely to report that providers spent enough time with their children (odds ratio = 0.34, 95% confidence interval = 0.17-0.68) compared to English speaking parents. There was no statistically significant difference found between Spanish-speaking parents with no need of interpretive services and English speaking parents. CONCLUSIONS: Among Spanish- versus English-speaking parents, differences in ratings of whether providers spent enough time with children during medical encounters appear to be explained, in part, by need for interpretive services. No other differences in ratings of provider communication were found. PMID- 15548102 TI - The involvement of low-income African American fathers in their children's lives, and the barriers they face. AB - OBJECTIVE: To examine the involvement of fathers in the lives of low-income African American 8-year-old children, and the barriers they face. METHODS: The sample was comprised of 117 fathers or father figures of 8-year-olds in families participating in a longitudinal study of child development and maltreatment. The men were asked a series of open-ended questions pertaining to their involvement in the children's lives. Their responses were audiotaped and transcribed. Major themes and subthemes were identified and coded on NVIVO software. RESULTS: The men conveyed a strong sense of commitment to the children, identifying many issues reported by white and middle class men, such as providing support and affection and teaching values and skills. They raised the need to protect the children and help take care of them when sick, some adding that they did not feel confident doing so. They saw discipline as one of their roles, but described this as difficult for them. The men faced challenges of not having financial resources, not living with the child, and lacking knowledge or skills. CONCLUSIONS: This group of fathers appears to be clearly committed to their children, despite significant challenges. There are a variety of ways that pediatricians can help facilitate their positive involvement in children's lives, and they may well contribute to the health and development of such high-risk children. PMID- 15548103 TI - Emergency department use and perceived delay in accessing illness care among children with Medicaid. AB - BACKGROUND: Children covered by Medicaid are at increased risk of emergency department (ED) utilization.Objective.-To examine whether an association exists between ED use and perceived delay in accessing acute care. DESIGN AND SETTING: Cross-sectional study. We used data from the Consumer Assessment of Health Plans Survey collected by Washington State Medicaid in 2000. We used multivariate logistic regression to determine the odds of any ED use, using a model that included whether the parent reported not always receiving illness care for the child as soon as wanted (perceived delay), age, gender, race/ethnicity, health status, presence of a special health care need, primary language, needing an interpreter, parental education, and having a regular provider. SUBJECTS: Parents of a random sample of children from 9 Medicaid managed care plans were surveyed if their children were <15 years old and enrolled at least 6 months. MAIN OUTCOME MEASURES: Any ED use in the past 6 months. RESULTS: The response rate was 56.3%, yielding 5142 subjects. Of children with illnesses in the previous 6 months, 69.1% of parents reported that their child always received care as soon as they wanted; 19.9%, 8.2%, and 2.8% reported usually, sometimes, and never, respectively. Not always (vs always) receiving illness care as soon as wanted was significantly associated with increased odds of any ED use (adjusted odds ratio: 1.79; 95% confidence interval: 1.35- 2.36). CONCLUSION: Children with managed care Medicaid are more likely to use the ED if parents perceive any delay in receiving illness care. PMID- 15548104 TI - Pediatric resident education about medical errors. AB - BACKGROUND: National organizations have called for patient safety curricula to help reduce the incidence of errors. Little is known about what trainees are taught about medical errors. OBJECTIVE: 1) To determine the amount and type of training that pediatric residents have about medical errors and 2) to assess pediatric chief resident knowledge about medical errors. METHODS: We surveyed chief residents from a national sample of 51 pediatric training programs by selecting every fourth program from the American Council on Graduate Medical Education list of accredited programs. The 21-item telephone survey was developed with patient safety specialists and piloted on several chief residents. It asked about patient-safety training sessions and awareness and knowledge about medical errors. RESULTS: The 51 chief residents helped teach 2176 residents, approximately one third of all pediatric residents. One third of programs had no lectures about medical errors and 23% did not have morbidity and mortality rounds. Sixty-one percent of respondents stated that outpatient medical errors were rarely discussed. Informal teaching was most often reported as the primary method for educating residents about medical errors. Although 58% of respondents did not know that a systemic change should be made in response to a medical error, 83% felt that residents are adequately trained to deal with a medical error. DISCUSSION: Pediatric resident education about medical errors varies widely. Attention by pediatric residency training programs to this important issue seems limited. PMID- 15548106 TI - Residency Review Committee (RRC) guidelines for pediatric residency training. PMID- 15548107 TI - Level of supervision of young children while in bathtubs. PMID- 15548110 TI - Single-strand-specific radiosensitization of DNA by bromodeoxyuridine. AB - The effects of bromodeoxyuridine (BrdUrd) substitution for thymidine on gamma-ray induced strand breakage were determined in single- and double-stranded oligonucleotides and double-stranded oligonucleotides containing a mismatched bubble region. BrdUrd does not sensitize complementary double-stranded DNA to gamma-ray-induced strand breakage, but it greatly sensitizes single-stranded DNA. However, when the BrdUrd is present in a single-stranded bubble of a double stranded oligonucleotide, the non-base-paired nucleotides adjacent to the BrdUrd as well as several unpaired sites on the opposite unsubstituted strand are strongly sensitized. The radiosensitization properties of BrdUrd result primarily from the electrophilic nature of the bromine, making it a good leaving group and leading to the irreversible formation of the uridine-yl radical (dUrd(.)) or the uridine-yl anion (dUrd(-)) upon addition of an electron. The radiolytic loss of the bromine atom is greatly suppressed in double-stranded compared to single stranded DNA. Thus we propose that the radiosensitization effects of bromouracil in vivo will likely be limited to single-strand regions such as found in transcription bubbles, replication forks, DNA bulges and the loop region of telomeres. Our results may have profound implications for the clinical use of bromodeoxyuridine (BrdUrd) as a radiosensitizer as well as for the development of targeted radiosensitizers. PMID- 15548111 TI - Attempts at correlation of the radiolytic species of irradiated solid-state captopril studied by multi-frequency EPR and HPLC. AB - The purpose of this study was to provide insight into the processes that occur after the irradiation of solid-state drugs. Electron paramagnetic resonance (EPR) experiments were performed at two different frequencies, X-band (about 9.5 GHz) and Q-band (about 34 GHz), to identify the radicals present in irradiated captopril. The results confirmed that an irradiated drug can trap several main radicals. Moreover, the radical composition varied as a function of the treatment. In addition, non-volatile final products were studied by liquid chromatography coupled to UV and to mass spectrometry (LC-MS). The variation of the radical composition did not influence the profile of the final products; this appears to indicate that, in the case of captopril, the trapped radicals observed by EPR are not the main precursors of the final products. Finally, high performance liquid chromatography data appear to indicate that radiosterilization of captopril is feasible. PMID- 15548112 TI - Enhancement of radiation cytotoxicity by UCN-01 in non-small cell lung carcinoma cells. AB - Thoracic ionizing radiation is a standard component of combined-modality therapy for locally advanced non-small cell lung cancer. To improve low 5-year survival rates (5- 15%), new strategies for enhancing the effectiveness of ionizing radiation are needed. The kinase inhibitor UCN-01 has multiple cell cycle effects, including abrogation of DNA damage-induced S- and G(2)-phase arrest, which may limit DNA repair prior to mitosis. To test the hypothesis that therapy induced cell cycle effects would have an impact on the efficacy of a combination of UCN-01 plus ionizing radiation, the cell cycle responses of the non-small cell lung cancer cell lines Calu1 (TP53-null) and A549 (wild-type TP53) to 2 Gy ionizing radiation were correlated with clonogenic survival after irradiation plus UCN-01. Irradiated cells were exposed to UCN-01 simultaneously and at 3-h increments after irradiation. In Calu1 cells but not A549 cells, sequence dependent potentiation of radiation by UCN-01 was observed, with maximal interaction occurring when UCN-01 was administered 6 h after irradiation. This coincided with the postirradiation time with the greatest depletion of cells from G(1). Abrogation of G(2) arrest was observed regardless of TP53 status. The role of TP53 was investigated using siRNA to achieve gene silencing. These studies demonstrated that radiation plus UCN-01 was more effective in cells with diminished TP53 activity, associated with a reduced G(1) checkpoint arrest. These studies indicate that simultaneous elimination of multiple DNA damage-induced checkpoints in G(1), S and G(2) may enhance the effects of radiation and that drug scheduling may have an impact on clinical efficacy. PMID- 15548113 TI - A novel anticancer ribonucleoside, 1-(3-C-ethynyl-beta-D-ribo pentofuranosyl)cytosine, enhances radiation-induced cell death in tumor cells. AB - 1-(3-C-Ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS106) is a newly developed anti-tumor agent that targets RNA synthesis. We report here that a low dose of ECyd induces radiosensitization of caspase-dependent apoptosis and reproductive cell death in cells of the gastric tumor cell lines MKN45 and MKN28 and murine rectum adenocarcinoma Colon26. Flow cytometry demonstrated that TAS106 induced the abrogation of the X-ray-induced G(2)/M checkpoint. Western blot analysis showed that X rays increased the expression of cyclin B1, phospho-Cdc2 and Wee1, whereas co-treatment with X rays and TAS106 decreased the expression of these cell cycle proteins associated with the G(2)/M checkpoint. Furthermore, TAS106 was shown to decrease the radiation-induced expression of survivin but not Bcl2 and BclX(L) regardless of TP53 status and cell type. Overexpression of wild type survivin in MKN45 cells inhibited the induction of apoptosis induced by co treatment with X rays and TAS106. These results suggest that TAS106 enhances X ray-induced cell death through down-regulation of survivin and abrogation of the cell cycle machinery. PMID- 15548114 TI - Neoplastic transformation in vitro after exposure to low doses of mammographic energy X rays: quantitative and mechanistic aspects. AB - The induction of neoplastic transformation in vitro after exposure of HeLa x skin fibroblast hybrid cells to low doses of mammography-energy (28 kVp) X rays has been studied. The data indicate no evidence of an increase in transformation frequency over the range 0.05 to 22 cGy, and doses in the range 0.05 to 1.1 cGy may result in suppression of transformation frequencies to levels below that seen spontaneously. This finding is not consistent with a linear, no-threshold dose- response curve. The dose range at which possible suppression is evident includes doses typically experienced in mammographic examination of the human breast. Experiments are described that attempt to elucidate any possible role of bystander effects in modulating this low-dose radiation response. Not unexpectedly, inhibition of gap junction intercellular communication (GJIC) with the inhibitor lindane did not result in any significant alteration of transformation frequencies seen at doses of 0.27 or 5.4 cGy in these subconfluent cultures. Furthermore, no evidence of a bystander effect associated with factors secreted into the extracellular medium was seen in medium transfer experiments. Thus, in this system and under the experimental conditions used, bystander effects would not appear to be playing a major role in modulating the shape of the dose-response curve. PMID- 15548116 TI - Changes in micronucleus frequency resulting from preirradiation of cell culture surfaces. AB - We have initiated a series of experiments to quantify the impact of environmental variables on the observed frequency of micronuclei in monolayer cultures. In this paper the influence of preirradiation of cell culture vessels on micronucleus formation in Chinese hamster ovary cells was examined. Dry cell culture vessels were preirradiated with 2 Gy of either alpha particles or X rays and immediately plated with nonirradiated cells. About 48 h later a group of randomly chosen containers was set aside, and the rest of the containers were exposed to a range of doses of X rays or alpha-particle radiation. Nonirradiated cells plated on previously irradiated cell culture surfaces manifested nearly as many micronuclei as the irradiated cells. In all experiments, preirradiation of the cell substrate (the culture dish) led to a significantly increased micronucleus frequency relative to unirradiated substrate. These results suggest that methods of cell culture vessel sterilization and the composition of cell attachment surfaces could be a confounding factor, particularly in low-dose experiments. PMID- 15548115 TI - Down-regulation of Betaig-h3 gene is involved in the tumorigenesis in human bronchial epithelial cells induced by heavy-ion radiation. AB - High-energy (HZE) heavy ions, when compared to low-LET radiation, are highly effective in inducing gene mutation, chromosomal aberrations and neoplastic transformation. However, the underlying molecular mechanisms are not clearly understood. We have recently shown that the down-regulation of Betaig-h3 expression is causally linked to the tumorigenic phenotype of papillomavirus immortalized human bronchial epithelial (BEP2D) cells treated with high-LET alpha particle radiation. Using the BEP2D cell culture system, a radiation-induced transformation model has been established by a single 60-cGy dose of (56)Fe heavy ion radiation. To determine whether the Betaig-h3 gene is involved in (56)Fe ion induced tumorigenesis, the expression levels of the Betaig-h3 gene in tumorigenic cell lines and the ability of in vivo tumor suppression through the reintroduction of the Betaig-h3 gene in tumorigenic cells were determined. We found that the expression level of this gene is markedly decreased in three tumorigenic cell lines ((56)FeT1-T3) compared with parental BEP2D cells. Ectopic expression of its cDNA in the (56)FeT2 tumorigenic cells significantly suppressed their tumorigenicity. Although biologically active TGFB1 is elevated in two of three tumorigenic cell lines, all these cell lines are resistant to the induction of Betaig-h3 expression by incubating the transformed cells with exogenous TGFB1 relative to control cells. Our data strongly suggest that down-regulation of Betaig-h3 expression results from the defect in the TGFB1 signaling pathway and plays a pivotal role in the tumorigenic process induced by (56)Fe heavy-ion radiation. PMID- 15548117 TI - Relative sensitivities of repair-deficient mammalian cells for clonogenic survival after alpha-particle irradiation. AB - The clonogenic survival of cells of the radiation-sensitive hamster cell lines irs1, irs2, irs3 and xrs5, representing different DNA repair pathways, was compared to that of their parent lines after alpha-particle irradiation. Measurements of nuclear area were made to calculate the probability of surviving a single alpha-particle traversal, the average number of lethal lesions per track and per unit dose, along with the "intrinsic radiosensitivity" of these cells, allowing for the potential of multiple lethal lesions per traversal. For all cell lines studied, alpha particles were found to be more biologically effective per unit absorbed dose than X rays at inducing cell inactivation. The repair deficient cells showed an enhanced sensitivity to alpha particles compared to their parent line, but the degree of enhancement was less than for X rays. The reduction in additional sensitivity for alpha-particle irradiation was shown not to be due predominantly to differences in cell geometry limiting the probability of a cell nucleus being traversed. The results suggest that both the nonhomologous end-joining pathway and to a lesser extent the homologous recombination repair pathway play a role in successful repair of alpha-particle induced damage, although a large proportion of damage is not repaired by either pathway. PMID- 15548118 TI - Induction of replication protein A in bystander cells. AB - The bystander effect is a biological phenomenon whereby cells not directly targeted by DNA-damaging agents elicit a response similar to that of targeted cells. Understanding the mechanisms underlying the bystander effect is important not only for radiation risk assessment but also for evaluation of protocols for radiotherapy of tumors. Identification of DNA repair and signal transduction proteins that are induced specifically in bystander cells may help in deducing the molecular mechanism(s) responsible for this complex phenomenon. With this objective, we have studied the expression of replication protein A (RPA), which is involved in various DNA metabolic activities such as replication, repair and recombination. We analyzed RPA expression by immunofluorescence and Western blot techniques in both gamma-irradiated primary human fibroblast cells and bystander cells that were recipients of conditioned growth medium harvested from gamma irradiated cell cultures. A two- to threefold induction of RPA was observed in bystander MRC5 cells treated with conditioned medium collected from gamma irradiated WI38 or MRC5 cells. Lack of induction of RPA in sham-manipulated MRC5 cells treated with irradiated medium alone (without cells) indicates that the signal elicited from the irradiated cells is responsible for induction of RPA in bystander cells. RPA was induced more effectively in bystander cells than in irradiated cells at the earliest time analyzed (30 min), and the RPA level declined to that of sham-treated control cells by 24 h after treatment. In addition to RPA, apurinic/apyrimidinic endonuclease (APE, a key enzyme of the base excision repair pathway) also showed enhanced expression in bystander cells. Our findings suggest that the induction of RPA and APE is due to a combination of DNA strand breaks and oxidized base lesions in the genomic DNA of bystander cells. PMID- 15548119 TI - Backscatter and dose perturbations for low- to medium-energy electron point sources at the interface between materials with different atomic numbers. AB - Electron backscatter at interfaces between dissimilar media can affect dosimetry and should be taken into consideration in radiotherapy and in radiobiology experiments. Backscatter dose perturbations depend upon factors such as electron energy, medium atomic number (Z), and distance from the interface. This study quantifies the backscatter dose factor (BSDF) for electron point sources of energy between 0.1 to 3 MeV in water at the interface with scattering materials ranging in Z from (13)Al to (79)Au. A Monte Carlo code that performs dose calculations for monoenergetic and continuous-spectrum electron sources was developed using EGSnrc transport routines. The BSDF was quantified in a parallel layers geometry (BSDF(1D)) and three-dimensional voxel geometry (BSDF(3D)). The BSDF(1D) near the interface increased up to 52% with decreasing energy from 3 to 0.1 MeV and increasing Z from 13 to 79. The analysis of the BSDF(3D) showed a significant dependence of the scattered electron angular distribution on Z and energy, with a decrease in isotropy going from high to low Z. This effect proves the importance of considering the correct geometry when quantifying the BSDF for electron sources, especially when the dimensions of the relevant dose-collecting volume are comparable with the CSDA range of the source. PMID- 15548120 TI - The microdosimetry of the (10)B reaction in boron neutron capture therapy: a new generalized theory. AB - The microdosimetry of (10)B thermal neutron capture reactions should be considered as an essential step to be followed before studying the radiobiological aspects of boron neutron capture therapy. The boron dose itself is insufficient as the only quantity used to describe the biological effectiveness of the (10)B reaction for two important reasons: the specific microdistribution that the (10)B carrier compound exhibits at the cellular level and the primarily stochastic nature of the energy deposition process, which influences the biological response to the particulate radiation. In this work, these two aspects are analyzed in detail and an innovative rigorous analytical framework is developed in the microdosimetry domain. This formalism provides the necessary microdosimetric tools for more precisely describing the (10)B dose distribution deposited in sensitive microscopic structures and offers improved approaches for analyzing the biological dose--effect relationship of (10)B reactions. PMID- 15548121 TI - Models for evaluating agents intended for the prophylaxis, mitigation and treatment of radiation injuries. Report of an NCI Workshop, December 3-4, 2003. AB - To develop approaches to prophylaxis/protection, mitigation and treatment of radiation injuries, appropriate models are needed that integrate the complex events that occur in the radiation-exposed organism. While the spectrum of agents in clinical use or preclinical development is limited, new research findings promise improvements in survival after whole-body irradiation and reductions in the risk of adverse effects of radiotherapy. Approaches include agents that act on the initial radiochemical events, agents that prevent or reduce progression of radiation damage, and agents that facilitate recovery from radiation injuries. While the mechanisms of action for most of the agents with known efficacy are yet to be fully determined, many seem to be operating at the tissue, organ or whole animal level as well as the cellular level. Thus research on prophylaxis/protection, mitigation and treatment of radiation injuries will require studies in whole animal models. Discovery, development and delivery of effective radiation modulators will also require collaboration among researchers in diverse fields such as radiation biology, inflammation, physiology, toxicology, immunology, tissue injury, drug development and radiation oncology. Additional investment in training more scientists in radiation biology and in the research portfolio addressing radiological and nuclear terrorism would benefit the general population in case of a radiological terrorism event or a large-scale accidental event as well as benefit patients treated with radiation. PMID- 15548122 TI - Comparisons of LET distributions for protons with energies between 50 and 200 MeV determined using a spherical tissue-equivalent proportional counter (TEPC) and a position-sensitive silicon spectrometer (RRMD-III). AB - Experiments have been performed to measure the response of a spherical tissue equivalent proportional counter (TEPC) and a silicon-based LET spectrometer (RRMD III) to protons with energies ranging from 50-200 MeV. This represents a large portion of the energy distribution for trapped protons encountered by astronauts in low-Earth orbit. The beam energies were obtained using plastic polycarbonate degraders with a monoenergetic beam that was extracted from a proton synchrotron. The LET spectrometer provided excellent agreement with the expected LET distribution emerging from the energy degraders. The TEPC cannot measure the LET distribution directly. However, the frequency mean value of lineal energy, y( )(f), provided a good approximation to LET. This is in contrast to previous results for high-energy heavy ions where y(-)(f) underestimated LET, whereas the dose-averaged lineal energy, y(-)(D), provided a good approximation to LET. PMID- 15548123 TI - Assessment of drug therapy management and the prevalence of heart failure in a managed care population with hypertension. AB - OBJECTIVES: To (1) determine the prevalence of heart failure (HF) and cardiovascular risk factors within a hypertensive managed care population, (2) measure blood pressure goal attainment in patients with concurrent HF and hypertension (HTN), and (3) assess the use of drug therapy for diabetic and nondiabetic patients with concurrent HF and HTN, particularly regarding the use of angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor blockers (ARBs). METHODS: Eligible patients were identified through a review of medical and pharmacy claims data from 10 managed care organizations (MCOs) and 2 specialty medical groups (4.6 million total members) from June 1998 through July 2001. From approximately 850,000 members in the claims database identified as hypertensive, 7,226 were randomly selected for medical chart review. Of these, 6,935 medical charts had a confirmed diagnosis of HTN but not HF, and 291 (4%) had confirmed HTN and HF. The study population--291 patients with HTN and HF- provided information on demographic characteristics, prevalence of cardiovascular risk factors and relevant comorbidities, and systolic and diastolic blood pressure. Current antihypertensive therapy prescription fill rate was evaluated using pharmacy claims. RESULTS: Patients with diagnoses of HTN and HF confirmed in the medical chart (N=291) were included in the present analysis. HF prevalence among hypertensive patients was 4% (291 of 7,226). Mean age of the study patients was 68.3 years, and 52.9% of the patients were female. Key cardiovascular risk factors included gender (men and postmenopausal women) (89.3%), age > 60 years (73.5%), hyperlipidemia (47.4%), and diabetes (38.8%). Of the total sample, only 30.1% of the diabetic (34 of 113) and 26.4% of the nondiabetic (47 of 178) patients with HF had their blood pressure controlled to the goal level of < 130/85 mm Hg recommended by the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure, the national guideline in effect at the time. Overall, 64.7% of HF patients for whom we had pharmacy claims were receiving an ACE inhibitor or an ARB. CONCLUSIONS: The study results indicate a deficit in the treatment of HTN among HF patients with and without diabetes, including failure to achieve blood pressure goals (< 130/85 mm Hg at the time of this study period). More aggressive quality improvement programs are necessary to educate providers and patients on the importance of treating blood pressure to nationally accepted goal using antihypertensives proven beneficial for hypertensive patients with HF. PMID- 15548124 TI - Severe sepsis in managed care: analysis of incidence, one-year mortality, and associated costs of care. AB - OBJECTIVE: To determine severe sepsis (SS) incidence, hospital mortality, 1-year mortality, and costs associated with care in a sample of enrollees in a nationally representative individual practice association (IPA)-network managed care organization (MCO). METHODS: This was a retrospective analysis of administrative claims data for commercial (not managed Medicare) members. We identified MCO members hospitalized for SS between July 1995 and December 1998. SS cases were identified by a combination of ICD-9-CM codes for infection and organ dysfunction. Enrollment information, physician, facility, and pharmacy claims were analyzed. Subjects with continuous enrollment were followed for 1 full year of observation. Costs were health plan payments to providers, after subtraction of member cost-share amounts. RESULTS: The incidence rate was 0.91 cases of SS per 1,000 enrollees, increasing with age. The mean age of SS patients was 50 years, and 53% were male. Approximately 63% received surgical intervention. Mortality was 21% during the first hospitalization and 36.1% at 1 year. During follow-up, 47.1% of survivors were rehospitalized. Mean index hospitalization length of stay and costs were 16 days and 26,820 dollars, with 1 year inpatient and outpatient costs totaling 48,996 dollars. Mean outpatient costs per survivor were 8,363 dollars, and mean per-patient-per-month (PPPM) outpatient costs were 906 dollars. Total follow-up costs including rehospitalization were similar for nonsurvivors compared with survivors (7,710 dollars versus 8,522 dollars, P=0.274), but PPPM costs were higher for nonsurvivors (1,760 dollars versus 699 dollars, P<0.001). CONCLUSIONS: Incidence, hospital, and 1-year mortality rates were lower in this population compared with literature reports and were associated with a lower average age in this managed care population. Mean SS hospitalization costs were high, and nearly one half of survivors required rehospitalization within 1 year. Study results suggest the need to evaluate SS interventions for improvement in health outcomes and cost outcomes, particularly in postsurgical patients. PMID- 15548125 TI - Cost-minimization analysis of once-weekly versus thrice-weekly epoetin alfa for chemotherapy-related anemia. AB - BACKGROUND: For individuals with chemotherapy-related anemia, the clinical effectiveness of epoetin alfa (EPO) dosed once weekly ([QW], 40,000 units per dose) has been demonstrated to be indistinguishable from that observed with thrice-weekly dosing ([TIW], 10,000 units per dose). Whether the advantage of less-frequent administration justifies the higher EPO dosage used in the weekly regimen in terms of overall cost of care is unknown. OBJECTIVE: To conduct a cost minimization analysis comparing QW and TIW EPO dosing from a societal perspective. METHODS: Direct and indirect medical cost data were calculated for a 16-week period for 2 large, prospective, multicenter, community-based studies. Costs measured included EPO, transfusions, laboratory tests, office visits, and opportunity cost of patient time. RESULTS: The average total costs in 2002 (first half) dollars were nearly equivalent across the 2 groups (QW: 9,204 dollars; 95% confidence interval [CI], 9,057 dollars-9,350 dollars. TIW: 9,265 dollars; 95% CI, 9,083 dollars-9,447 dollars. P=0.60). QW incurred mean drug acquisition costs that were 23% higher (QW: 6,725 dollars; 95% CI, 6,611 dollars-6,838 dollars. TIW: 5,474 dollars; 95% CI, 5,350 dollars-5,598 dollars. P<0.001). However, QW patients can avoid the resource use and time cost associated with 2 additional office visits incurred each week (QW: 592 dollars [583 dollars-600 dollars]; TIW: 1,709 dollars [1,678 dollars-1,740 dollars]; P<0.001). Transfusion and laboratory test costs were slightly higher in the TIW group (QW: 1,888 dollars [1,837 dollars-1,940 dollars]; TIW: 2,082 dollars [2,020 dollars-2,144 dollars]; P<0.001). CONCLUSION: Total anemia treatment costs over a 16-week period with EPO QW were similar to those of TIW dosing. In the absence of cost differences between regimens, the noneconomic advantages of less-frequent dosing intervals should make weekly dosing increasingly attractive to patients, clinicians, and payers. PMID- 15548126 TI - Longitudinal analysis of the use of etanercept versus infliximab determined from medical chart audit. AB - OBJECTIVE: To describe the dosing of etanercept and infliximab for the treatment of rheumatoid arthritis (RA). METHODS: Adult patients with a diagnosis of RA who were treated with either etanercept or infliximab between 1999 and 2002 were selected from 16 rheumatology practices in the western and southeastern United States. Patients with a terminal illness or those receiving a tumor necrosis factor (TNF)-alpha inhibitor for an indication other than RA were excluded. Data were collected through a review of the patient medical records. Data collected on each patient included demographics, concurrent disease-modifying antirheumatic drug therapy, TNF-alpha inhibitor dose, frequency, duration of TNF-alpha inhibitor therapy, and discontinuation of TNF-alpha inhibitor therapy. RESULTS: A total of 244 patients were included in the evaluation (etanercept only [n=128; 52%], infliximab only [n=89; 36%], both [n=27; 11%]). The mean age of these patients was 55.1+/-13.3 years, 54.9+/-13.5 years, and 52.8+/-14.0 years, respectively; the mean duration of RA was 13.3 +/- 8.8 years, 13.4+/-8.0 years, and 14.0 +/- 9.9 years, respectively. Female patients constituted 70% of the sample. Health maintenance organization insurance was the most common form of medical insurance (45.8%), followed by Medicare (22.3%). The mean duration of follow-up for etanercept and infliximab treatment was 29.3+/-14.1 months and 14.8+/-6.9 months, respectively. Among patients who were still receiving therapy at the time of review, the mean initial and last etanercept doses were 25.0 mg versus 25.8 mg (P=0.16); the mean initial and last infliximab doses were 3.38 mg/kg versus 4.51 mg/kg (P<0.001). CONCLUSION: The dosing of etanercept and infliximab therapy was consistent with the approved labeling of both medications. PMID- 15548127 TI - Managed care market perspectives on the over-the-counter availability of statins. AB - OBJECTIVE: In April, May, and June 2003, we conducted market research with decision makers from 3 payer segments to determine their perspectives on the potential introduction of statins to the U.S. over-the-counter (OTC) market. METHODS: We selected a convenience sample of survey participants based upon prominence in the market, membership size, and by willingness to participate in our study. The 12 participating managed care organizations (MCOs) cover approximately 100 million lives. The 4 pharmacy benefit managers (PBMs) cover approximately 200 million lives. The 3 large employers (one employer withheld quantitative results) provide medical coverage to nearly 1.4 million employees, both actives and retirees. Each survey participant received a verbal description of an OTC statin and a proposed patient self-management system. We asked each participant a series of questions to obtain opinions on a number of issues related to the potential introduction of an OTC statin. RESULTS: Our research findings can be summarized in the following key conclusions: (1) MCO representatives generally view OTC statins as a low-risk and beneficial addition to drug therapy options; (2) payer policies will continue to support access to prescription statins with no change in policy following introduction of an OTC option; (3) several of the MCOs and 75% of the PBMs anticipate a sharp, short term increase in plan costs as a consequence of OTC statin availability; (4) survey participants believe that consumer or member reaction will be mixed and that consumer advertising and physician education will be important; (5) MCOs and PBMs are eager for involvement but cautious about partnerships with pharmaceutical companies; and (6) the details of any OTC statin offering will lay the foundation for its success. CONCLUSIONS: Based upon interviews conducted from April through June 2003, key decision makers from 4 PBMs, 12 MCOs, and 3 large employers generally considered the introduction of an OTC statin as a low risk and a beneficial addition to drug therapy. Most believed that increased awareness would result in an initial increase in plan costs, but long-term savings would accrue through improved care and availability of lower-cost OTC options for low to-moderate-risk patients. The key concern is how to help patients gain enough knowledge and comfort to manage their own cholesterol therapy safely and successfully. PMID- 15548128 TI - Market withdrawal of Vioxx: is it time to rethink the use of COX-2 inhibitors? PMID- 15548129 TI - Spending money to save money--cardiovascular disease management. PMID- 15548130 TI - David and Goliath--employer takes on health care cost management. PMID- 15548131 TI - Categorizing patients from medical claims data--the influence of GIGO. PMID- 15548132 TI - Guiding principles for effective electronic messaging. PMID- 15548133 TI - Dose consolidation can be an efficient intervention. PMID- 15548134 TI - Nucleotide specificity of HIV-1 reverse transcriptases with amino acid substitutions affecting Ala-114. AB - Ala-114, together with Asp-113, Tyr-115 and Gln-151, form the pocket that accommodates the 3'-OH of the incoming dNTP in the HIV-1 RT (reverse transcriptase). Four mutant RTs having serine, glycine, threonine or valine instead of Ala-114 were obtained by site-directed mutagenesis. While mutants A114S and A114G retained significant DNA polymerase activity, A114T and A114V showed very low catalytic efficiency in nucleotide incorporation assays, due to their high apparent K(m) values for dNTP. Discrimination between AZTTP (3'-azido 3'-deoxythymidine triphosphate) and dTTP was not significantly affected by mutations A114S and A114G in assays carried out with heteropolymeric template/primers. However, both mutants showed decreased susceptibility to AZTTP when poly(rA)/(dT)16 was used as substrate. Steady-state kinetic analysis of the incorporation of ddNTPs compared with dNTPs showed that substituting glycine for Ala-114 produced a 5-6-fold increase in the RT's ability to discriminate against ddNTPs (including the physiologically relevant metabolites of zalcitabine and didanosine), a result that was confirmed in primer-extension assays. In contrast, A114S and A114V showed wild-type ddNTP/dNTP discrimination efficiencies. Discrimination against ribonucleotides was not affected by mutations at position 114. Misinsertion and mispair extension fidelity assays as well as determinations of G-->A mutation frequencies using a lacZ complementation assay showed that, unlike Tyr-115 or Gln-151 mutants, the fidelity of HIV-1 RT was not largely affected by substitutions of Ala-114. The role of the side-chain of Ala-114 in ddNTP/dNTP discrimination appears to be determined by its participation in van der Waals interactions with the ribose moiety of the incoming nucleotide. PMID- 15548135 TI - Factor VIIa stimulates endothelin-1 synthesis in TNF-primed endothelial cells by activation of protease-activated receptor 2. AB - The mechanisms linking prothrombotic changes to endothelial dysfunction and accelerated atheroma formation have yet to be fully defined. Expression of TF (tissue factor) on the endothelium is potentially an initiating event as binding and activation of FVII (factor VII) can result in thrombosis. Although PAR2 (protease-activated receptor-2) is expressed on vascular endothelium, its precise physiological significance and mechanism of activation have yet to be defined. In the present study, we investigated whether PAR2 can be activated by FVIIa (activated FVII) and induce ET-1 (endothelin-1) synthesis. In bovine aortic endothelial cells pretreated with TNF (tumour necrosis factor-alpha) to increase TF expression, FVIIa stimulated ET-1 synthesis via activation of PAR2. Although FX (factor X) alone was inactive, this response was enhanced by using FVII and FX in combination. Inhibition of the proteolytic activity of FVIIa abolished the response. The PAR2 agonist peptide SLIGKV also enhanced ET-1 release on TNF pretreated cells. The response to FVIIa was inhibited by a PAR2 antagonist peptide FSLLRY. Inhibition of the p38 MAPK (mitogen-activated protein kinase) reduced PAR2 expression and the ET-1 response. In summary, FVIIa can stimulate ET 1 synthesis in endothelial cells by activating PAR2, demonstrating a potential link between thrombotic processes and endothelial cell dysfunction. PMID- 15548136 TI - PAK and other Rho-associated kinases--effectors with surprisingly diverse mechanisms of regulation. AB - The Rho GTPases are a family of molecular switches that are critical regulators of signal transduction pathways in eukaryotic cells. They are known principally for their role in regulating the cytoskeleton, and do so by recruiting a variety of downstream effector proteins. Kinases form an important class of Rho effector, and part of the biological complexity brought about by switching on a single GTPase results from downstream phosphorylation cascades. Here we focus on our current understanding of the way in which different Rho-associated serine/threonine kinases, denoted PAK (p21-activated kinase), MLK (mixed-lineage kinase), ROK (Rho-kinase), MRCK (myotonin-related Cdc42-binding kinase), CRIK (citron kinase) and PKN (protein kinase novel), interact with and are regulated by their partner GTPases. All of these kinases have in common an ability to dimerize, and in most cases interact with a variety of other proteins that are important for their function. A diversity of known structures underpin the Rho GTPase-kinase interaction, but only in the case of PAK do we have a good molecular understanding of kinase regulation. The ability of Rho GTPases to co ordinate spatial and temporal phosphorylation events explains in part their prominent role in eukaryotic cell biology. PMID- 15548138 TI - "Intimate surgery" of the female: a word of caution. PMID- 15548137 TI - Bromodomain analysis of Brd2-dependent transcriptional activation of cyclin A. AB - Cyclin A is regulated primarily through transcription control during the mammalian cell cycle. A dual mechanism of cyclin A transcriptional repression involves, on the one hand, promoter-bound inhibitory complexes of E2F transcription factors and RB (retinoblastoma) family proteins, and on the other, chromatin-directed histone deacetylase activity that is recruited to the cyclin A promoter early in the cell cycle in association with these RB proteins. This dual regulation maintains transcriptional silence of the cyclin A locus until its transcription is required in S-phase. At that time, RB family members dissociate from E2F proteins and nucleosomal restructuring of the locus takes place, to permit transcriptional activation and resultant S-phase progression to proceed. We have identified a double bromo-domain-containing protein Brd2, which exhibits apparent 'scaffold' or transcriptional adapter functions and mediates recruitment of both E2F transcription factors and chromatin-remodelling activity to the cyclin A promoter. We have shown previously that Brd2-containing nuclear, multiprotein complexes contain E2F-1 and -2. In the present study, we show that, in S-phase, they also contain histone H4-directed acetylase activity. Overexpression of Brd2 in fibroblasts accelerates the cell cycle through increased expression of cyclin A and its associated cyclin-dependent kinase activity. Chromatin immunoprecipitation studies show that Brd2 is physically present at the cyclin A promoter and its overexpression promotes increased histone H4 acetylation at the promoter as it becomes transcriptionally active, suggesting a new model for the dual regulation of cyclin A. PMID- 15548139 TI - Examiner's unique possibilities to catalyze women's empowerment during a pelvic examination. PMID- 15548140 TI - Association between fertility drugs and gynecologic cancers, breast cancer, and childhood cancers. AB - Ovulation-inducing drugs have been widely used for various types of infertility since the beginning of 1960s and their use increases day by day parallel to the success achieved in fertility treatment. However, the researches performed in the last two decades have begun to discuss about the safety of these drugs and the risks associated with their use. Especially, the potential neoplastic effects of these drugs are increasingly questioned. The studies have discussed whether there is an association between the exposure to ovulation-inducing drugs and the incidence of various cancers. Moreover, several studies have been performed to reveal whether there is an increased risk of childhood cancers in children conceived after fertility treatment. The point we reached through the available data is that the risk of breast, uterine and invasive ovarian cancers is not increased, but the risk of borderline ovarian tumors might increase after such a therapy. The risk of cancer has been found similar for children conceived after fertility treatment and those conceived naturally. It should also be kept in mind that cancers are overdiagnosed in infertile women population because of the close medical surveillance, which may also contribute to the early detection of cancers. Although it is still early to state the last words on this topic, the possible association should be addressed when obtaining an informed consent before starting treatment. PMID- 15548141 TI - Well-being and sense of coherence during pregnancy. AB - BACKGROUND: Our knowledge of how a woman as an individual experiences, evaluates and copes with pregnancy is limited. The concept of 'sense of coherence' as described by Antonovsky may be an important factor in how people deal with stressful situations in life. The aim of this study is to describe pregnant women's perception of well-being during pregnancy and after delivery and to explore its relationship to their sense of coherence. METHODS: Participants (n = 177) were recruited from four Maternal Health Care Centers (MHCCs) in the Stockholm area. Three standard instruments were used in the study: the Sense of Coherence (SOC) scale, the Health Index (HI) and the Hospital Anxiety and Depression (HAD) scale. These instruments were filled in at pregnancy weeks 10-12 and 34-36 and also 8 weeks after delivery. RESULTS: One hundred and twenty women completed the study. At weeks 34-36 the pregnant women scored significantly worse for well-being than at weeks 10-12 and after delivery. A strong predictor for well-being is the women's degree of SOC. With regard to anxiety and depression no differences were found between the measurements. Neither demographic data such as age or financial status nor degree of complications influenced the results. CONCLUSIONS: Our study shows that the SOC scale measures the capacity to cope with the unforeseeable process that child-bearing still implies today. The SOC scale questionnaire and the HI can complement the midwife's subjective evaluation of the pregnant woman's need for psychosocial support during pregnancy. If this is correct, the SOC scale and the HI could be of great value in clinical work within maternal health care. PMID- 15548142 TI - Prostacyclin, thromboxane A and the effect of low-dose ASA in pregnancies at high risk for hypertensive disorders. AB - BACKGROUND: The aim of this study was to investigate the prostanoid production in pregnancies at high risk for hypertensive disorders, and the effect of low-dose acetylsalicylic acid (ASA) on prostanoids. MATERIAL AND METHODS: Ninety women with a bilateral notching in uterine arteries screened by Doppler ultrasound at 12-14 gestational weeks were randomized to the ASA (0.5 mg/kg/day) or placebo group. Forty-three women in both groups were followed up throughout the pregnancy. Urine samples were taken at baseline, and at 24-26 and 32-34 weeks of gestation to determine the urinary 11-dehydrothromboxane B(2) (u-11-dehydro TxB(2)) and 2,3-dinor-6-keto-prostaglandin F(1alpha) (u-2,3-dinor-6-keto PGF(1alpha)), the metabolites of thromboxane A(2) and prostacyclin, respectively. RESULTS: In the pregnancies with pregnancy-induced hypertension (PIH) before 37 gestational weeks, the 2,3-dinor-6-keto-PGF(1alpha)/11-dehydro-TxB(2) ratio did not increase as much as in other pregnancies (P = 0.028). In the placebo group pregnancies with preeclampsia had significantly lower 2,3-dinor-6-keto PGF(1alpha) (P = 0.019) at 12-14 weeks of gestation compared to other pregnancies. In the placebo group the 2,3-dinor-6-keto-PGF(1alpha)/11 dehydroTxB(2) ratio remained unchanged throughout the pregnancy, with no significant difference between pregnancies with a normal or an adverse outcome. In the ASA group the 2,3-dinor-6-keto-PGF(1alpha)/11-dehydro-TxB(2) ratio increased (P < 0.001, early vs. midpregnancy). Again, the changes were similar in pregnancies with a normal or an adverse outcome. CONCLUSION: The balance of prostacyclin and thromboxane A(2) shifted in an unfavorable direction in pregnancies complicated by PIH. ASA had a favorable effect on the prostanoids. PMID- 15548143 TI - Major congenital heart disease in Northern Norway: shortcomings of pre- and postnatal diagnosis. AB - BACKGROUND: Congenital heart disease (CHD) is a significant cause of perinatal mortality and morbidity worldwide. Prenatal detection rate of CHD remains low in most European countries and a substantial proportion of infants with serious heart disease are diagnosed only after discharge from hospital after birth. Earlier recognition of treatable abnormalities may improve the perinatal outcome. The purpose of this study was to evaluate the incidence, pre- and postnatal diagnosis and outcome of major CHD in two northern-most counties of Norway. MATERIALS AND METHODS: All cases of major CHD were identified by reviewing the records of all fetal and neonatal echocardiographies and all perinatal autopsies performed during the study period. Data were obtained on maternal age, associated risk factors, time of diagnosis, type of CHD and outcome. RESULTS: Ninety-one fetal and 52 neonatal echocardiographies and 77 perinatal autopsies were performed during the 2-year study period. There was a total of 5658 births during this period. Twenty-five cases of major CHD were identified. Six of them were diagnosed prenatally, 18 after birth, and one additional case was identified from the perinatal autopsy register. The incidence of major CHD in Northern Norway was 4.4 per 1000 births. The prenatal detection rate was 24% (6/25). In 39% (7/18) of cases diagnosed postnatally the diagnosis was made following a second admission after the baby has been initially discharged from hospital after birth. The mortality among liveborn infants with major CHD was 17.4% (4/23). CONCLUSION: Despite almost universal prenatal ultrasound screening and at least one neonatal examination before discharge, the diagnosis of major CHD is made after a relevant delay in a substantial proportion of cases. PMID- 15548144 TI - Pregnancy outcomes in unselected singleton pregnant women with an increased risk of first-trimester Down's syndrome. AB - BACKGROUND: The purpose of this study was to assess outcomes in pregnancies with a positive screen of first-trimester combined test (nuchal translucency, pregnancy-associated plasma protein-A and free beta-human chorionic gonadotropin). METHODS: Using a cut-off level of 1 in 270, 216 (7.1%) women had a positive screen. Among them, 187 delivered their babies in our hospital and the adverse outcomes, such as spontaneous abortion, intrauterine fetal demize, preterm prelabor rupture of the membranes, preterm labor, intrauterine growth restriction, gestational hypertensive disorders, placenta previa, chromosomal abnormalities and fetal structural anomalies, were identified and compared with the 2097 women who screened negative for Down's syndrome. RESULTS: Pregnancies with a positive screen had a significantly higher risk of adverse outcomes than those with negative screens (30.5% versus 15.3%; odds ratio 2.4; p < 0.001), especially for those complicated by spontaneous abortion (odds ratio 11.4; p < 0.05) and placenta previa (odds ratio 4.3; p < 0.05). CONCLUSIONS: Besides fetal chromosomal abnormalities and structural abnormalities, pregnancies with a positive screen for Down's syndrome in the first-trimester had a significantly higher incidence of subsequent adverse obstetric outcomes. PMID- 15548145 TI - Births and perinatal health of infants among women who have had silicone breast implantation in Finland, 1967-2000. AB - BACKGROUND: Potential problems with breast implants have been widely discussed, but few data exist on the childbearing and offspring of women with implants. The purpose of this study was to investigate the occurrence and conditions of pregnancies of women who have had cosmetic breast implantation (exposed women), and the health of their newborns. METHODS: Women who had breast implants for cosmetic reasons in the period 1967-1999 (n = 2236) were identified from hospital surgical records. The births of the exposed women were identified through record linkage to the Population Register. The perinatal health of the infants was studied by the data in the Medical Birth Register in 1987-1999. For each birth to an exposed woman, 20 control mothers who gave birth in the same year were chosen randomly from the Medical Birth Register. Differences in mothers' background characteristics were adjusted by logistic regression. RESULTS: The women had received their first cosmetic breast implants at young ages (mean 31 years). By year 2000, 26% of the exposed women had one or more children. Half of these women had not had a liveborn child before getting implants. Of the 1661 exposed women who had not (yet) had children, 32% were less than 35 years of age at the end of follow-up. The women had children at a mean of 4.7 years after the implants. Some of the perinatal health indicators suggested poorer health and others better health for infants of exposed women, but only transfers to other hospitals and lower birthweight among infants of exposed multipara were statistically significant. CONCLUSIONS: The study shows that pregnancy and infant health are relevant considerations with regard to breast implants; further studies on implants are needed. PMID- 15548146 TI - Two-stage ultrasonography in screening for fetal anomalies at 13-14 and 18-22 weeks of gestation. AB - BACKGROUND: The aim of this study was to assess the value of two-stage screening by ultrasonography in detecting selected major fetal anomalies in a low-risk obstetric population. METHODS: In a defined geographic area, 4789 consecutive low risk pregnant women participated in screening by two-stage ultrasonography as part of routine maternal care. The examinations were usually performed by specially trained midwives at 13-14 and 18-22 weeks of gestation. Of the women, 4073 had both scans, 440 had the early one only, and 276 the late scan only. Pregnancy outcomes were ascertained from obstetric and pediatric records, and the data were supplemented with information from the national birth and malformation registries. RESULTS: Of the 4855 fetuses, 33 (0.7%) had major structural defects considered detectable by ultrasonography. Of these, six (18%) were identified at the early scan, and an additional 10 (30%) at the late scan, yielding a total sensitivity of 48% for the two-stage screening. Twenty offspring had chromosomal abnormalities; 10 were identified by increased nuchal translucency at the early scan, one additional one (by hydronephrosis) at the late scan, and the remaining nine at birth. CONCLUSIONS: In a low-risk population, first-trimester scanning is useful in finding fetuses with chromosomal anomalies, but a second-trimester scan is needed for other types of defects. The sensitivity of routine screening by midwives for fetal structural defects in a general obstetric population remains lower than that reported by specialized centers. PMID- 15548147 TI - Reproductive factors and extreme levels of maternal serum alpha-fetoprotein: a population-based study. AB - BACKGROUND: Levels of maternal alpha-fetoprotein (AFP) are increased during multiple gestations and preeclampsia but little is known regarding AFP levels in relation to other reproductive factors. Consequently, the objective of this work was to describe the possible relationship between AFP levels during pregnancy and maternal age at birth, maternal age at first birth, parity, time since previous birth and gender of the offspring. METHODS: Based on national registries we obtained the reproductive history on a population-based cohort of 44 227 women who had serum AFP levels determined in gestational weeks 14-21 and whose present and previous pregnancies resulted in live-born singletons. RESULTS: Many previous births and an interval of less than 2 years since last birth were significantly associated with extremely low levels of AFP in the mother. However, age at first birth and age at present pregnancy did not influence the AFP level. Women who gave birth to a girl had AFP levels that were 5%[95% confidence interval (CI) 4 6%] lower than those of women who had a boy. Adjustment for birthweight did not significantly affect the estimate. CONCLUSIONS: Low serum AFP levels in pregnancy are significantly correlated with high parity and with a short interval between births. The significantly lower levels of AFP in women who gave birth to girls could indicate a possible gender-specific regulatory mechanism. PMID- 15548148 TI - Subclinical abnormalities of glucose metabolism in Brazilian women with a history of gestational diabetes mellitus. AB - BACKGROUND: To evaluate insulin release and insulin sensitivity in women with prior gestational diabetes mellitus (GDM) to gain a better understanding of type 2 diabetes pathogenesis. METHODS: GDM women were individually matched for age, body mass index, and waist/hip ratio with those who were normal glucose tolerant in a previous pregnancy (NGT). All women presented with normal glucose tolerance. Twenty pairs were submitted to the oral glucose tolerance test (OGTT) with plasma glucose, insulin, and C-peptide determinations. Of the 20 pairs, 18 participated in hyperglycemic (10.0 mmol/l) clamp experiments with frequent plasma glucose and insulin determinations, allowing us to calculate first- and second-phase insulin release and the insulin sensitivity index. GDM and NGT women were compared using Student's t-test, the Mann-Whitney U-test, Friedman's non-parametric test, and the two proportion test for independent groups. RESULTS: GDM women showed higher glycosylated hemoglobin values; at OGTT, they showed late insulin peak with increased plasma insulin levels only during the second hour, and a similar plasma C-peptide response despite a higher plasma glucose curve; during hyperglycemic clamp procedures, they showed similar biphasic insulin release and insulin sensitivity index. Considering that a woman with previous GDM had a defect in insulin release and/or insulin sensitivity, if its magnitude was at least 25% lower than that of the matched NGT woman, 43.8% showed impairment of first-phase insulin release and 55.6% insulin resistance. CONCLUSIONS: GDM women showed some degree of glucose intolerance. It is therefore necessary to follow them for a longer time. PMID- 15548149 TI - Housing conditions, perceived stress, smoking, and alcohol: determinants of fetal growth in Northwest Russia. AB - BACKGROUND: Social determinants of health have become important in countries of the former Soviet Union during the time of transition. AIM: This article estimates the influence of living conditions, smoking, alcohol, and maternal stress on fetal growth indices in an urban Russian setting. METHODS: All pregnant women registered at antenatal care centres in Severodvinsk (Northwest Russia) and their infants comprised the study base (n = 1399). Multivariate linear regression analysis was applied to quantify the effects of the studied factors on birth weight and the ponderal index (PI) adjusted for maternal education, age, occupation, marital status, parity, pre-pregnancy weight, paternal employment, gestational age, and others. RESULTS: Infants of smoking mothers were 126 g lighter compared with non-smokers [95% confidence interval (CI): -198; -54]. On average, birth weight decreased 27 g per cigarette smoked during pregnancy. Living in shared apartments, living in crowded housing situations, and perceived stress were associated significantly with birth weight loss: -89 g (95% CI: -153; -25), -82 g (95% CI: -136; -28), and -61 g (95% CI: -116; -7), respectively. A positive association between maternal alcohol consumption and birth weight was found. Living with parents was associated positively with both birthweight and PI. Infants whose fathers consumed more than 100 ml of absolute alcohol per week were thinner at birth compared with those of non-drinking and moderate drinking fathers. CONCLUSIONS: Poor housing conditions, maternal stress, smoking, and alcohol consumption are independent determinants of fetal growth in Northwest Russia. PMID- 15548150 TI - Subsequent pregnancy outcome after conservative treatment of a previous cesarean scar pregnancy. AB - BACKGROUND: To assess pregnancy course and outcome after conservative treatment of a cesarean scar pregnancy. METHODS: During an 8-year period, 15 cases of cesarean scar pregnancies were diagnosed at our institution. Seven of the 14 patients for whom we successfully preserved the uterus became pregnant within 3 years after termination of the scar pregnancy. The year of diagnosis, conservative method and gestational age for these five patients were recorded. Delivery method, time interval between the scar pregnancy and subsequent pregnancy, and maternal and neonatal outcome were evaluated. RESULTS: Seven pregnancies (eight live and one dead baby) were noted. The mean interval between the ectopic pregnancy and subsequent pregnancy was 13.3 months (range 0-34 months). One patient, who became pregnant 3 months after the scar pregnancy was found, suffered uterine rupture at 38.3 weeks' gestational age. Two patients with placental accrete, and one of them who continued the existing intrauterine twin pregnancy after transvaginal sono-guided aspiration of the scar pregnancy received a cesarean hysterectomy at 32 weeks of gestation. The remaining four pregnancies were uneventful, followed by early cesarean sections at 36 weeks. CONCLUSION: The results of this first series of seven subsequent pregnancies after conservative treatment of scar pregnancies are promising. An early cesarean section before over-extension of the uterus and spontaneous labor can help to prevent uterine rupture. Placenta accrete is another severe morbidity of these patients in addition to uterine rupture. Thus a cesarean hysterectomy may be the choice of treatment. PMID- 15548151 TI - Low plasma levels of oxidized low density lipoprotein in preeclampsia. AB - BACKGROUND: Markers of lipid peroxidation are commonly used to assess oxidative stress in preeclampsia. The aim of this study was to assess the concentration of oxidized low density lipoprotein (oxLDL), a novel marker for lipid peroxidation, and that of the thiobarbituric acid reactive substances (TBARS) in the pathogenesis of severe preeclampsia and to investigate the influence of gestational age on these parameters. METHOD: Plasma levels of oxLDL and TBARS were assayed in women with severe preeclampsia (n = 40), normotensive pregnant controls matched for gestational age (n = 24) and normotensive pregnant controls at full term (n = 16). RESULTS: Women with preeclampsia showed lower oxLDL levels (mean +/- SE) than matched controls (181 +/- 12 vs. 219 +/- 14; p = 0.027), whereas no differences were found for the TBARS concentration (3.8 +/- 0.6 vs. 3.7 +/- 0.4). When women with preeclampsia were compared to control women at full term, TBARS were elevated (3.8 +/- 0.6 vs. 1.5 +/- 0.2; p = 0.01). However, in women with normotensive pregnancy TBARS were also lower in full-term control pregnancy compared to early third-trimester values (p < 0.0001). CONCLUSION: Plasma TBARS decreased during the third trimester of pregnancy, underlining the importance of matching for gestational age when studying markers of lipid peroxidation in pregnant women. Women with preeclampsia had lower plasma levels of oxLDL compared to gestational age-matched controls, indicating that oxLDL could be a marker for preeclampsia. PMID- 15548152 TI - Association between maternal age and the likelihood of a cesarean section: a population-based multivariate logistic regression analysis. AB - BACKGROUND: A majority of studies examining the relationship between advancing maternal age and the likelihood of cesarean section (CS) use data from regional samples or from a limited number of medical institutions. This study uses population-based data from Taiwan to explore the relationship between maternal age and the likelihood of a CS. METHODS: The National Health Insurance Research Database (NHIRD) on registries of medical facilities and board-certified physicians and monthly claim summaries for inpatients were used. In total, 502 524 singleton deliveries were included in the study. Multivariate logistic regressions were performed with the presence of CS as the dependent variable and maternal age (<20, 20-29, 30-34 and >34 years) as the independent variable. The study controlled for maternal indications, institution characteristics, maternal requests and attending physician characteristics. RESULTS: CS rates for the age groups <20, 20-29, 30-34 and >34 years were 17.7, 27.4, 37.4 and 47.5%, respectively. The regression analyses consistently showed that the likelihood of a CS significantly increased with advancing maternal age within each category of complication after adjusting for medical institution characteristics and characteristics of the attending physician. CONCLUSIONS: This study found that, after adjusting for maternal indications, and healthcare institution and physician characteristics, there was a significant relationship between advancing maternal age and an increased likelihood of a CS. This finding, together with the high CS rate of 32.1% in Taiwan, one of the highest reported in the world today, highlights an imperative need to devise interventions to reduce the frequency of CSs. PMID- 15548153 TI - Infertility and celiac disease: do we need more than one serological marker? AB - OBJECTIVES: Celiac disease (CD) prevalence is higher in women with infertility. Our study aims were to evaluate the prevalence of undiagnosed CD in Arab infertile women and to explore the usefulness of using more than one serological marker in the diagnostic screening for CD in this population. METHODS: Women with unexplained infertility (n = 192) and age-matched healthy controls (n = 210) were prospectively enrolled. Serum was tested for human tissue transglutaminase antibodies (TTG), antiendomysial antibodies (EMA), and immunoglobulin A. Intestinal biopsy was offered to women with positive serology or immunoglobulin A (IgA) deficiency. RESULTS: CD was diagnosed in five infertile women (2.65%) and in one control (0.5%) (p = 0.11). Gastrointestinal complaints were present in 60% (three of five) of women with CD and 11.8% (22 of 187) of women without CD (p = 0.017). Anemia was reported in 80% of infertile women with CD and 4.8% of infertile women without CD (p = 0.0001). CONCLUSIONS: Undiagnosed CD is prevalent in Arab infertile women as well as in Arab women in general. CD in Arab infertile women is frequently associated with gastrointestinal complaints and anemia. EMA testing is sufficient in suspected cases. PMID- 15548154 TI - Sexual behavior during the first eight weeks after legal termination of pregnancy. AB - BACKGROUND: The aim of the study was to describe sexual symptoms and resumption of sexual activity during the first 8 weeks after unrestricted first-trimester termination of pregnancy (TOP). METHODS: Questionnaires 2 and 8 weeks after legal first-trimester abortion in 1327 women. RESULTS: The 8-week questionnaire was returned by 941 (71%). Overall, 51.0% of the women recommenced coitus within 2 weeks after TOP. This figure was significantly higher among women aged 18-24 years (60.6%) than among women in higher age groups (41.7-47.8%). At 8 weeks follow-up coitus was reported by 86.9%, ranging from 65.8% of women not in a steady relationship to 93.7% of women in a relationship (p < 0.001), and age related differences were insignificant. Long duration of vaginal bleeding, medical abortion and need for reevacuation of the uterus diminished the resumption frequency at 2 weeks follow-up, but 8 weeks after termination postabortion complications did not affect resumption. At 8 weeks follow-up 15.3% reported diminished sexual desire and 6.0% reported diminished orgasm ability comparing to prepregnancy level. Of the sexually active women 6.2% did not use sufficient contraception 8 weeks after termination. CONCLUSION: Coitus is recommenced shortly after termination of pregnancy but not always with sufficient contraception. Legal abortion is associated with few adverse effects on sexual function. PMID- 15548155 TI - Young women with genital prolapse have a low collagen concentration. AB - BACKGROUND: Genital prolapse is a common and handicapping form of pelvic floor dysfunction. To explain its genesis as a result of endopelvic connective tissue weakness, the collagen state was analyzed in women with and without genital prolapse. METHODS: Punch biopsies from the paraurethral ligaments were obtained during the operation from 22 women undergoing surgery for genital prolapse. As controls, similar biopsies were taken from 13 women who underwent gynecologic surgery for other benign reasons. Collagen concentration as hydroxyproline and its extractability by pepsin digestion were studied in relation to age by multiple regression, two-way anova, Levene's test, and Student's t-test. Histological examination was also performed. RESULTS: Women, younger than 53 years, with genital prolapse had a 30% lower collagen concentration than age matched controls, which reached significance, P = 0.01. The extractability by pepsin digestion, an indicator of cross-links in the collagen molecule, did not significantly differ between groups. It did, however, decrease significantly with age in both prolapse patient and control groups. Morphology supported these findings with a less-dense extracellular matrix composition subepithelially in genital prolapse compared to a healthy control. CONCLUSION: For the first time, we show that young women with genital prolapse have a decreased collagen concentration, suggesting a different organization of the endopelvic connective tissue extracellular matrix. Furthermore, these alterations differ from those earlier found in younger women with stress urinary incontinence. PMID- 15548156 TI - Oral contraceptives and liver hemangioma: a case-control study. AB - OBJECTIVE: To study whether there is an epidemiological association between liver hemangiomas and oral contraception use, as may be suggested by clinical observations of liver hemangioma growth during pregnancy and under estrogen administration. METHODS: A case-control study of 40 women with liver hemangiomas, as diagnosed by imaging studies [ultrasonography (US) or computed tomography (CT) and nuclear scan], and 109 age-matched control with normal studies. The women were interviewed with respect to their menstrual, reproductive and oral contraception use history. RESULTS: The liver hemangioma and control groups did not differ significantly with regard to their menstrual or reproductive history. Ever oral contraception use was reported by 30% of the cases and 27% of controls. The odds ratio (OR) was 1.1 [95% confidence interval (CI) 0.52-2.60] for ever use, 1.64 (95% CI 0.37-7.13) for initiation before the age of 20, and 0.62 (95% CI 0.16-2.42) for use duration of less than 1 year. On the multivariate analysis only the women's age emerged as a predictor for liver hemangiomas (OR 3.1; 95% CI 1.16-8.96). CONCLUSIONS: In this study liver hemangiomas were not associated with menstrual, reproductive and oral contraception use history. PMID- 15548157 TI - Postoperative infections and antibiotic prophylaxis for hysterectomy in Sweden: a study by the Swedish National Register for Gynecologic Surgery. AB - AIM: The purpose of the study was to evaluate the current use of antibiotic prophylaxis, the rate of postoperative infections, and risk factors for postoperative infections in patients undergoing elective hysterectomy for non malignant pathology. METHODS: The study was performed during the time period July 2000 to 1 January 1 2003, using data from the Swedish National Register for Gynecologic Surgery, covering 26 of 49 clinics and approximately 50% of the Swedish population. During this time period, 3267 women undergoing elective hysterectomy for non-malignant pathology (mainly dysfunctional bleeding and fibromas) and with complete data on the use of antibiotic prophylaxis and postoperative follow up were included. A postoperative infection was considered to have developed if patients or the physician reported an infection related to the surgical site, together with reported use of antibiotic treatment. RESULTS: Among the 3267 patients, 341 (10.4%) were considered to have developed a postoperative infection. In cases where antibiotic prophylaxis had not been given, total abdominal hysterectomies and vaginal hysterectomies were associated with the highest frequency for postoperative infections P < 0.05 and P < 0.01, respectively. Notably, also women undergoing subtotal hysterectomy reported significantly lower rates of postoperative infections if prophylactic antibiotics had been given, P < 0.05. Risk factors for postoperative infection were non-use of antibiotic prophylaxis, hospital, age less than 40 years, and BMI > 30 kg/m(2). CONCLUSIONS: The study results have indicated the need of prospective clinical trials on antibiotic prophylaxis for patients undergoing supravaginal abdominal hysterectomy. Also, a need to further evaluate the current prophylactic antibiotic protocols used for obese patients and younger women undergoing any kind of hysterectomy is warranted. PMID- 15548158 TI - Trust and confirmation in a gynecologic examination situation: a critical incident technique analysis. AB - BACKGROUND: Gynecologic examination is a common measure in reproductive health care. Many women experience the examination as a more or less negative event, with shortcomings in the examiner's behavior. The aim of the study was to describe, in terms of critical incidences, women's experiences concerning the personnel's behavior in the situation of gynecologic examination. METHODS: The informants were strategically chosen and consisted of 30 Swedish women between the ages of 18-82 years old. The data collection method was qualitative research interviews analyzed by critical incident technique. RESULTS: The result consisted of 30 subcategories, five categories, and two main areas - trust and confirmation. The personnel enabled trust when they promoted participation, created confidence, and were supportive. The opposite behavior contributed to the lack of trust. Confirmation described behavior that confirmed, respectively, did not confirm the women. This was shown through the presence or lack of respect and engagement. CONCLUSION: The personnel's positive behavior enabled trust and confirmed the women as individuals, while negative behavior was decisive in an unfavorable way. A complexity of patterns of knowing in nursing was identified. Participation through information that contributed to trust was important and amounted to one fourth of the incidents in the material. Respect and engagement, which confirmed the women, facilitated a positive caring relationship. The examination situation can be improved through reflection of the personnel's own behavior and further research about women's own experiences. PMID- 15548159 TI - GnRH antagonists in poor responders. PMID- 15548160 TI - Recurrent thromboembolism in ovarian cancer. PMID- 15548161 TI - Pyoderma gangrenosum as a cause of spontaneous vulvovaginal ulceration. PMID- 15548162 TI - Expectant management of a cornual pregnancy followed up by serial transvaginal color power Doppler angiography and serum beta human chorionic gonadotropin levels. PMID- 15548163 TI - Extreme hypertrophy of the labia minora. PMID- 15548164 TI - Reduction of maternal mortality--by prevention or cure? PMID- 15548165 TI - International medical school graduates in American medicine: is there the "dark side of the moon"? PMID- 15548166 TI - Bone mineral density changes in pregnancy. PMID- 15548171 TI - The role of topical calcineurin inhibitors in atopic dermatitis. AB - For more than five decades, topical corticosteroids and emollients have been the mainstay of therapy for atopic dermatitis. However, the potential for side effects limits the clinical utility of corticosteroids in providing long-term disease control. With a unique mode of action that differs from that of corticosteroids, the steroid-free topical calcineurin inhibitors (TCIs), tacrolimus ointment and pimecrolimus cream, provide skin-selective treatment that targets key factors involved in the pathogenesis of this chronic disease. An extensive series of clinical trials involving more than 16,000 patients with predominantly moderate to severe atopic dermatitis in tacrolimus studies and over 2000 patients with primarily mild to moderate disease in pimecrolimus studies has shown that both TCIs provide effective and well-tolerated treatment for atopic dermatitis. Randomized controlled trials have demonstrated that tacrolimus is superior to conventional hydrocortisone-based regimens and does not cause skin atrophy or other steroidal side-effects. Both tacrolimus and pimecrolimus prevent disease flares and provide progressive and sustained disease improvement with long-term therapy. These and other clinical benefits of TCIs are discussed, together with the safety profiles of tacrolimus and pimecrolimus and their use in clinical practice. In addition, this review summarizes findings from the many trials carried out with these agents and outlines how TCIs can provide long-term treatment and control of a chronic skin disease that may persist for years. PMID- 15548172 TI - Teduglutide ([Gly2]GLP-2) protects small intestinal stem cells from radiation damage. AB - Glucagon-like peptide-2 and its dipeptidyl peptidase (DP-IV) resistant analogue teduglutide are trophic for the gastrointestinal epithelium. Exposure increases villus height and crypt size and results in increased overall intestinal weight. As these effects may be mediated through stimulation of the stem cell compartment, they may promote intestinal healing and act as potential anti mucositis agents in patients undergoing cancer chemotherapy. A study was initiated to investigate the protective effects of teduglutide on the murine small intestinal epithelium following gamma-irradiation using the crypt microcolony assay as a measure of stem cell survival and functional competence. Teduglutide demonstrated intestinotrophic effects in both CD1 and BDF1 mouse strains. In BDF1 mice, subcutaneous injection of GLP-2 or teduglutide (0.2 mg/kg/day, b.i.d.) for 14 days increased intestinal weight by 28% and resulted in comparable increases in crypt size, villus height and area. Teduglutide given daily for 6 or 14 days prior to whole body, gamma-irradiation significantly increased crypt stem cell survival when compared with vehicle-treated controls. The mean levels of protection over a range of doses provided protection factors from 1.3 to 1.5. A protective effect was only observed when teduglutide was given before irradiation. These results suggest that teduglutide has the ability to modulate clonogenic stem cell survival in the small intestine and this may have a useful clinical application in the prevention of cancer therapy-induced mucositis. PMID- 15548173 TI - In vivo effects of interleukin-17 on haematopoietic cells and cytokine release in normal mice. AB - In order to gain more insight into mechanisms operating on the haematopoietic activity of the T-cell-derived cytokine, interleukin-17 (IL-17) and target cells that first respond to its action in vivo, the influence of a single intravenous injection of recombinant mouse IL-17 on bone marrow progenitors, further morphologically recognizable cells and peripheral blood cells was assessed in normal mice up to 72 h after treatment. Simultaneously, the release of IL-6, IL 10, IGF-I, IFN-gamma and NO by bone marrow cells was determined. Results showed that, in bone marrow, IL-17 did not affect granulocyte-macrophage (CFU-GM) progenitors, but induced a persistant increase in the number of morphologically recognizable proliferative granulocytes (PG) up to 48 h after treatment. The number of immature erythroid (BFU-E) progenitors was increased at 48 h, while the number of mature erythroid (CFU-E) progenitors was decreased up to 48 h. In peripheral blood, white blood cells were increased 6 h after treatment, mainly because of the increase in the number of lymphocytes. IL-17 also increased IL-6 release and NO production 6 h after administration. Additional in vitro assessment on bone marrow highly enriched Lin- progenitor cells, demonstrated a slightly enhancing effect of IL-17 on CFU-GM and no influence on BFU-E, suggesting the importance of bone marrow accessory cells and secondary induced cytokines for IL-17 mediated effects on progenitor cells. Taken together, these results demonstrate that in vivo IL-17 affects both granulocytic and erythroid lineages, with more mature haematopoietic progenitors responding first to its action. The opposite effects exerted on PG and CFU-E found at the same time indicate that IL-17, as a component of a regulatory network, is able to intervene in mechanisms that shift haematopoiesis from the erythroid to the granulocytic lineage. PMID- 15548174 TI - 3-O-methylfunicone, a secondary metabolite produced by Penicillium pinophilum, induces growth arrest and apoptosis in HeLa cells. AB - 3-O-Methylfunicone (OMF) is a secondary metabolite produced by the soil fungus Penicillium pinophilum which has cytostatic properties. The aim of this study was to investigate the mechanisms by which such properties are exerted, with special reference to any anti-proliferative and apoptotic potential, on HeLa cells. OMF treatment caused about 44% inhibition of cell growth after 24 h, and modifications in the tubulin fibre organization. In addition, a significant increase in p21 mRNA expression and a decrease in cyclin D1 and Cdk4 mRNA expression resulted at the same time. Apoptosis induction was demonstrated by the annexin V assay, cytofluorimetric analysis of the DNA content of the sub-G1 fraction and DNA laddering. Taken together, our data showed that the compound inhibits proliferation of HeLa cells by several mechanisms, such as disruption of tubulin fibres, cell cycle arrest and apoptosis induction. The capacity of the compound to affect the cell cycle and to modulate apoptosis is indicative of a potential for the development of a new agent for cancer chemotherapy. PMID- 15548175 TI - Extracellular pressure stimulates colon cancer cell proliferation via a mechanism requiring PKC and tyrosine kinase signals. AB - Pressure in colonic tumours may increase during constipation, obstruction or peri operatively. Pressure enhances colonocyte adhesion by a c-Src- and actin cytoskeleton-dependent PKC-independent pathway. We hypothesized that pressure activates mitogenic signals. METHODS: Malignant colonocytes on a collagen I matrix were subjected to 15 mmHg pressure. ERK, p38, c-Src and Akt phosphorylation and PKCalpha redistribution were assessed by western blot after 30 min and PKC activation by ELISA. Cells were counted after 24 h and after inhibition of each signal, tyrosine phosphorylation or actin depolymerization. RESULTS: Pressure time-dependently increased SW620 and HCT-116 cell counts on collagen or fibronectin (P < 0.01). Pressure increased the SW620 S-phase fraction from 28 +/- 1 to 47 +/- 1% (P = 0.0002). Pressure activated p38, ERK, and c-Src (P < 0.05 each) but not Akt/PKB. Pressure decreased cytosolic PKC activity, and translocated PKCalpha to a membrane fraction. Blockade of p38, ERK, c-Src or PI-3 K or actin depolymerization did not inhibit pressure-stimulated proliferation. However, global tyrosine kinase blockade (genistein) and PKC blockade (calphostin C) negated pressure-induced proliferation. CONCLUSIONS: Extracellular pressure stimulates cell proliferation and activates several signals. However, the mitogenic effect of pressure requires only tyrosine kinase and PKCalpha activation. Pressure may modulate colon cancer growth and implantation by two distinct pathways, one stimulating proliferation and the other promoting adhesion. PMID- 15548176 TI - Improved efficiency of EBV transformation of B-lymphocytes. PMID- 15548177 TI - Internal thoracic arterial grafts evaluation by multislice CT scan: a preliminary study. AB - BACKGROUND AND AIM: The internal thoracic artery (ITA) has a better long-term patency than saphenous veins, and anastomosis between ITA and the left anterior descending artery (LAD) represents the "gold-standard" of surgical myocardial revascularization. The aim of this study is to evaluate the multidetector multislice CT Scan (MCTS) as a means of postoperative evaluation of ITA coronary artery bypass grafts. METHODS: Twenty-eight patients having been operated on for coronary artery bypass with ITA during a 6-months period, benefited, 7 days after surgery, from a patency and anastomotic site control of ITA with a MCTS associated with cardiac gating (Light Speed, General Electric, USA). RESULTS: Internal thoracic artery bypasses are visualized perfectly on all their courses, with possibility of 3D reconstructions, showing the relationship between cardiac cavities and the arterial bypasses. The anastomotic site on the LAD was, in selected cases, perfectly visualized. Sequential bypasses with left ITA are well visualized as well as T or Y right-to-left ITA grafts. However, surgical clips create some image artefacts. CONCLUSIONS: The postoperative control of ITAs are possible by MCTS with a satisfactory resolution. This makes it possible to check the patency of ITAs, their course on the heart surface, and the location and quality of anastomosis with a noninvasive reproductive method. PMID- 15548178 TI - Side effects of cardiopulmonary bypass: what is the reality? AB - Despite many years of clinical and experimental research, the contribution of cardiopulmonary bypass (CPB) and cardioplegic arrest to morbidity and mortality following cardiac surgery remains unclear. This is due, in part, to lack of suitable control group against which bypass and cardioplegic arrest can be compared. The recent success of beating heart coronary artery bypass grafting has, however, for the first time, provided an opportunity to compare the same operation, in similar patient groups, with, or without CPB and cardioplegic arrest. CPB is associated with an acute phase reaction of protease cascades, leucocyte, and platelet activation that result in tissue injury. This is largely manifest as subclinical organ dysfunction that produces a clinical effect in those patients that generate an excessive inflammatory response or in those with limited functional reserve. The contribution of myocardial ischemia/reperfusion, secondary to aortic cross-clamping, and cardioplegic arrest, to the systemic inflammatory response and wider organ dysfunction is unknown, and requires further evaluation in clinical trials. PMID- 15548179 TI - Total arterial off-pump versus on-pump coronary revascularization: comparison of early outcome. AB - BACKGROUND AND AIM: To assess differences in the early outcome after complete arterial myocardial revascularization with (ONCAB) or without cardiopulmonary bypass (OPCAB). METHODS: Out of 870 consecutive CABG procedures 58 OPCAB and 91 ONCAB patients receiving exclusive arterial grafts were analyzed. OPCAB patients had more single-vessel (p < 0.0001), less triple-vessel (p < 0.0001) or left main disease (p = 0.0021), higher angina class (p = 0.003), unstable angina (p < 0.0001) or previous PTCAs (p < 0.0001). RESULTS: ONCAB was associated with longer operations (182.5 +/- 38 vs. 147 +/- 56 min; p = 0.0001) and more anastomoses/patient (3.2 +/- 1 vs. 2 +/- 0.9; p < 0.0001), but incomplete revascularization was similar in both groups (11% vs. 17%; p = ns). ITA use was identical, whereas single left internal thoracic artery (LITA) use (25.9% vs.1%; p < 0.0001) and LITA jump anastomoses (10.3% vs. 7.7%; p < 0.0001) were more frequent in OPCAB. Radial artery (RA) use (89% vs. 46.6%; p < 0.0001) and RA jump anastomoses (57.1% vs. 12.1%; p < 0.0001) were more frequent in ONCAB. Mortality, arrhythmias, cerebro-vascular accidents (CVA), and renal failure were similar, but ventilatory support shorter (8.8 +/- 11.8 vs. 15.6 +/- 9.4 h; p < 0.0001) and cardiac enzyme release smaller (p < 0.0001) after OPCAB with a trend toward less myocardial infarction (1.7% vs. 7.7%; p = 0.12) and low output (1.7% vs. 8.8%; p = 0.089), and more respiratory complications (10.3% vs. 2.2%; p = 0.056). CONCLUSIONS: Arterial OPCAB patients have less extensive CAD, but more severe symptoms. Early outcome is similar concerning mortality, arrhythmias, CVA, renal failure, or ICU and hospital stay, but with shorter ventilatory support and lower cardiac enzymes with a trend toward lower myocardial infarction and low output, but higher respiratory complication rates after OPCAB. PMID- 15548180 TI - The pneumatic tourniquet technique for endoscopic radial artery harvest. AB - BACKGROUND: Endoscopic vessel harvest is viewed as a safe, reliable, and cost effective method for greater saphenous vein removal. As the radial artery has more recently become a popular conduit choice in coronary artery revascularization, we describe herein an endoscopic technique for radial artery harvest in our first 50 patients as an alternative to the more traditional open technique. METHODS: From November 2001 to July 2002, 54 radial arteries were harvested endoscopically in 50 patients utilizing the VasoView Endoscopic Vessel Harvesting System, an Esmark bandage, and a pneumatic cuff tourniquet. RESULTS: No patients experienced symptoms of vascular compromise, or signs of infection in the donor arm. No adjunctive procedures were required during the vessel harvest, i.e., conversion to open technique. All radial arteries were successfully removed with endoscopic technique and 53 of the 54 radial arteries were successfully used as bypass conduits. Although the quality of the radial artery harvested remains subjective, we found less vasospasm than in our experience with the open technique. Thirty-day follow-up revealed no readmissions, no cardiac ischemic complications, no significant complications with the donor arm, and an excellent cosmetic result. CONCLUSIONS: Endoscopic harvest of the radial artery with the tourniquet technique may offer advantages over the more traditional open technique. PMID- 15548181 TI - Bypass to the distal right coronary artery using in situ gastroepiploic artery. AB - OBJECTIVE: We investigated the clinical outcomes after coronary artery bypass grafting (CABG) using the in situ gastroepiploic artery (GEA) exclusively bypassing to the distal right coronary artery (RCA). METHODS: Between, 1991 and 2002, 1020 consecutive patients (788 male and 232 female with a mean age of 63.6 +/- 9.7) underwent CABG to the distal coronary artery using in situ GEA in the Shin-Tokyo Hospital Group. Their perioperative and follow-up data were retrospectively analyzed. RESULTS: The in-hospital morbidity rate was 15.1% and the mortality rate was 0.6%. Postoperative myocardial infarction associated with GEA graft failure occurred in two cases. Among the survivors, follow-up was completed in all patients with a mean period of 4.7 +/- 2.4 years. Actuarial 3- and 5-year angina or intervention-free rates were 93.6% and 89.9%, respectively. Actuarial 3- and 5-year survival rates were 95.8% and 91.6%, respectively. Postoperative angiography was performed on 482 patients within 1 year, 115 patients with 3 years, and in 87 patients more than 4 year after surgery. The GEA patency rates at 1, 3, and 5-year were 95.7%, 90.2%, and 85.7%, respectively. These patency rates were compared with other graft materials (the saphenous vein n = 291, radial artery n = 186, and right internal thoracic artery n = 23) used for bypass to the distal RCA, and we found no significant differences (p = 0.29). CONCLUSION: The perioperative and clinical remote results of GEA grafting were satisfactory. The angiographic patency rates of GEA to the distal RCA was not inferior to the saphenous vein graft in this study. PMID- 15548182 TI - Harvesting radial artery and neurologic complications. AB - BACKGROUND: Determination of the incidence, mechanisms, and diagnosis of hand complications after radial artery (RA) harvesting in coronary surgery (CABG). METHODS: The study group (RA group) includes 54 patients who underwent RA harvesting in CABG operation. The control group (noRA group) consists of 131 patients who underwent CABG without the use of RA graft. The average follow-up time was 16.36 +/- 5.13 months. The patients were examined clinically, (a) for motor function abnormalities associated with radial and median nerve damage and (b) for sensory abnormalities, and the function of radial nerve was determined by eliciting the brachioradialis reflex. They answered in a formal scripted questionnaire to elicit symptoms and clinical points attributable to nerve damage during RA harvest, such as hand weakness, thumb weakness, sensation abnormalities on the back and on the palm side of the forearm, hand numbness, hand-reversible paresis or forearm infection postoperatively, and any other upper limb abnormality. RESULTS: Of the patients in the RA group, 34.09% reported left-hand abnormality after operation. On the other hand, in the noRA group left-hand abnormality was reported in 18.68% of patients. In the RA group sensation abnormality was reported in 34.09% of patients and thumb weakness alone was reported in 6.82% of patients. There was a statistically significant difference between the two groups. Low EuroSCORE was the predicting factor for motor abnormalities. CONCLUSIONS: More knowledge has been added about the neurologic complications after RA harvesting lately. We demonstrated the rate of motor and sensory abnormality, the potential mechanisms of these complications caused by surgical trauma or devascularization, and any predictive factors of complications. Optimal surgical techniques to avoid the damage of the responsible nerves are recommended. PMID- 15548183 TI - What is the mortality and recuperative difference of bilateral versus single thoracic artery coronary revascularization in patients with reoperation or over 80 years of age? AB - BACKGROUND: We examined whether bilateral internal thoracic artery revascularization (BITA) is safe for reoperative coronary revascularization (reop CABG) or primary CABG at age > or = 80 (CABG > or = 80 yrs) as, these two groups are thought to be at higher risk for death or sternal infection. METHODS: We analyzed 329 such patients between January 1, 1993 and March 31, 2002. These are subgroups of 3200 prospectively New York State risk stratified patients for BITA or SITA (single internal thoracic artery revascularization) of equivalent preoperative risk. In 37/39, BITA > or = 80 since 1996 (1996 > or = 80) the microscope was used and the free right internal thoracic artery was anastomosed to the aorta. Long-term survival was analyzed by Kaplan-Meier curves and in particular among the 36 patients between 1996 and 1997, who were operated five and six years ago (1997 > or = 80). CONCLUSIONS: Mortality and recuperative difference of BITA versus SITA in the reop CABG and CABG > or = 80 years are negligible, as there was no significant difference in hospital mortality, sternal infections, LOS, or Kaplan-Meier survival curves and average long-term survival. However BITA appears to have long-term advantage over SITA in the newer period and beyond 48 months (1996-1997 > or = 80). PMID- 15548184 TI - The history and development of direct coronary surgery without cardiopulmonary bypass. AB - The history of direct myocardial revascularization without cardiopulmonary bypass dates to 1961 in the dawn of coronary artery surgery. With the introduction and development of techniques of extracorporeal circulation around the same time, beating heart surgery was largely abandoned. Over the subsequent decades, cardiopulmonary bypass and electromechanical cardioplegic arrest became popular as means of revascularization in a bloodless and motionless field. While coronary artery surgery on the arrested heart remained undisputed for decades, myocardial revascularization on the beating heart was pursued by a few pioneering surgeons around the world, based on the belief that coronary revascularization could be performed equally well without the detrimental effects of cardiopulmonary bypass and electromechanical arrest. Various concepts and techniques developed during the 1980s by these pioneers enabled minimally invasive coronary surgery to be performed in the early 1990s. This break from the mainstream allowed selective myocardial revascularization using a minimal incision and no cardiopulmonary bypass to develop and constructed a base for future extensive revascularizations off-pump. With the subsequent explosion of new techniques for coronary exposure and myocardial stabilization, complete revascularization without cardiopulmonary bypass became possible with consistent results. Emerging from the preview of only a few surgeons just a decade ago, off-pump surgery is currently one of the accepted modalities for complete myocardial revascularization worldwide. This paradigm shift in the approach to myocardial revascularization has led to exiting new future possibilities, such as beating heart totally endoscopic coronary artery surgery. PMID- 15548185 TI - Hospital outcome after aorta-radial versus internal thoracic artery-radial artery grafts. AB - BACKGROUND: We researched our data to determine whether use of radial artery (RA) led to similar hospital morbidity as use of pedicled internal thoracic artery (ITA) with vein grafts. We also investigated if use of RA, different RA operative techniques, or number of inflow grafts were predictors for hospital outcome. METHOD: Retrospectively the hospital outcome of the first 512 patients with RAs (RA group) was compared with 108 matched patients with left ITA (LITA) and vein grafts (LITA control group). Two subgroups of RA operative techniques were further analyzed: 327 patients with RA directly from aorta (aorta-RA group), and 185 patients with RA from ITA, as a composite graft, (ITA-RA group). RESULTS: Hospital outcome of the RA group was similar to that of the LITA control group. When all ischemic events (IE) were grouped together, univariate analysis showed that aorta-RA group resulted in less IE than the ITA-RA group (2.1% versus 5.9%, respectively, p = 0.025). Number of inflow grafts did not influence IE. Multivariate analysis, however, did not show that technique of proximal RA anastomosis or number of inflow grafts were predictors for IE. CONCLUSIONS: Hospital outcome after the use of the RA is similar to that of LITA with vein grafts. Univariate analysis shows less IE after direct aorta-RA anastomoses, but multivariate analysis did not show that technique of proximal RA anastomosis and number of inflow grafts are important predictors for hospital outcome. PMID- 15548186 TI - Both leaflet preservation during mitral valve replacement: modified anterior leaflet preservation technique. AB - BACKGROUND: Satisfactory results of bileaflet preserving mitral valve replacement (MVR) had forced several institutes to preserve both leaflets during MVR. Modifications were required to prevent the preserved tissue from interfering with prosthetic valve function, to implant an adequate size of valve and to prevent left ventricle outflow tract (LVOT) obstruction. MATERIALS AND METHODS: Conventional MVR was performed to 51 patients (group 1) and bileaflet preserving MVR was performed to 43 patients (group 2). Mitral anterior leaflet incised from the middle of the leaflet to mitral annulus without chordal injury in group 2 patients. Sutures were placed through the mitral annulus first and then passed from the bottom to the tip of anterior leaflet. Posterior leaflet was also preserved. Prosthetic valve was put down into the mitral annulus and sutures were ligated. Excessive anterior leaflet tissue was attached to left atrial wall. RESULTS: Cross-clamping time was 45 +/- 5.33 minutes versus 61.32 +/- 4.43 minutes (p = 0.0001) and total cardiopulmonary bypass time was 60.80 +/- 4.44 minutes versus 80.55 +/- 3.65 minutes (p = 0.0001) in groups 1 and 2, respectively. Inotropy requirement was higher in group 1 (p = 0.0058). When compared with preoperative values postoperative left ventricle ejection fraction (LVEF) increased both at rest (from 52.74% +/- 3.88% to 62.86% +/- 3.18%, p = 0.0001) and during exercise (from 53.16% +/- 3.16% to 64.11% +/- 2.46%, p = 0.0001) in bileaflet preserving MVR group. But in conventional MVR group LVEF decreased postoperatively both at rest (from 51.45% +/- 4.27% to 48.27% +/- 3.35%, p = 0.0001) and during exercise (from 54.47% +/- 7.36% to 42.96% +/- 3.58%, p = 0.0001). CONCLUSION: Leaflet preserving MVR operation not only improves the left ventricular performance but also reduces the mortality and morbidity after MVR. LVEF increases both at rest and during exercise. Risk of LVOT obstruction can be completely eliminated with our simple technique. PMID- 15548187 TI - Evaluation of the mitral valve leaflet morphology after mitral valve reconstruction with a concept "coaptation length index". AB - BACKGROUND: In clinical settings, information on morphology of mitral valve leaflet after mitral valve reconstruction is limited. METHODS: Between January 1996 and June 2000, 36 underwent mitral valve repair for mitral regurgitation (MR). The etiology of mitral insufficiency was prolapse, dilated annulus, and ischemic. Ring annuloplasty was performed in all cases. Mitral valve short-axis dimension (MVd), vertical distance between annular line and closing point (Vd), coaptation length (CL), coaptation length index (CL/MVd) were measured by the two dimensional transesophageal echocardiography for the present 11 cases. RESULTS: In 11 cases, residual MR, using a scale from 0 to 4, was 0 in 5 patients, 1 in 4 patients, 2 in 2 patients whose etiology of regurgitation was cardiomyopathy. MVd and Vd decreased significantly (38.7 +/- 6.2 to 27.0 +/- 5.6 mm, 10.1 +/- 7.7 to 6.5 +/- 4.6 mm, respectively). CL and CLI increased significantly (6.4 +/- 2.4 to 11.6 +/- 4.6 mm, 0.16 +/- 0.06 to 0.44 +/- 0.21, respectively). Among those index, only CLI have a statistically significant negative correlation with the degree of residual MR. CONCLUSION: The mitral valve ring annuloplasty produce the morphologic change of mitral apparatus, especially the increase of CLI, which may be one of the main factors in regulation of regurgitation. PMID- 15548188 TI - A technique for infusion of cardioplegic solution in coronary artery bypass with aortic regurgitation. AB - BACKGROUND: Complications such as heart failure due to insufficient cardioplegia may develop in on-pump coronary artery bypass (CAB) with mild-to-moderate aortic regurgitation (AR). A technique for administration of cardioplegic solution was carried out to avoid such complications. METHODS AND RESULTS: Cardiopulmonary bypass was established. After aortic cross-clamping, cardioplegic solution was administered from aortic root. Because complete cardiac arrest was not rapidly achieved, the aortic root was incised. Three cusps of the aortic valve were sutured. The aorta was closed; cardioplegic solution was administered from the aortic root. Then, cardiac arrest was rapidly achieved. After distal anastomosis of quadruple bypass was completed, the suture of the cusps was removed. There was no exacerbation of AR due to this method compared to the preoperative state. CONCLUSION: When off-pump coronary artery bypass is impossible and retrograde cardioplegia cannot be performed for a certain reason, this method may be set to one of the choices. PMID- 15548189 TI - Diffuse bullous emphysema of the lung: conservative resection with a local application of a biological glue. AB - A middle-aged man suffered a recurrent right pneumothorax with a prolonged air leak due to diffuse bullous emphysema of the lung. On December 19, 2001, he underwent a successful conservative resection of the bullae with a local application of a gelatin-resorcin-formalin biological glue. PMID- 15548190 TI - Surgical treatment of postinfarction left ventricular pseudoaneurysm. AB - In this report, we present two cases who had left ventricular pseudoaneurysm, resection, and repair was done successfully. The repair was performed with the aid of cardiopulmonary bypass. The neck of the pseudoaneurysm was closed with autologous fresh pericardial patch. The surgical treatment of ventricular pseudoaneurysm depends upon their origin, size, and local extension. PMID- 15548191 TI - A large left atrial myxoma. PMID- 15548192 TI - Contemporary trends in aortic valve surgery: a single centre 10-year clinical experience. AB - The purpose of this study is to present a comprehensive profile of the trends in aortic valve replacement at a single institution over the past decade. Prospectively collected data concerning 873 patients undergoing aortic valve replacement (AVR), with and without coronary artery bypass grafting (CABG), were analysed. The patients were divided into three time periods: period I, (1990 to 1993); period II, (1994 to 1996); and period III, (1997 to 2000). Actuarial survival of AVR patients with and without CABG at 7 years was 82.9 +/- 2.4% and 79.1 +/- 3.3% (p = 0.17), respectively. Actuarial survival at 7 years for stentless, mechanical, and stented valve patients were 89.5 +/- 2.7%, 85.5 +/- 2.8%, and 76.0 +/- 3.2%, respectively. There was a significant difference in survival between the stentless and stented valve groups (p = 0.014). Age (63.8 +/ 12.9 yrs, 66.2 +/- 11.0 yrs, 67.9 +/- 10.3 yrs; p = 0.01), the incidence of peripheral vascular disease (5.1%, 10.8%, 16.6%; p = 0.001), and the extent of coronary artery disease necessitating CABG (34.0%, 38.8%, 41.0%; p = 0.05) have increased significantly in the later time period. However, operative mortality has remained constant (4.7%, 4.8%, 4.5%; p = 0.9). Moreover, perioperative complications have decreased significantly (27.4%, 18.0, 16.0%; p = 0.001). Multivariate analysis identified more recent time period as independent protective factor for early mortality and morbidity (period I, RR 1.00; period II, RR 0.47; period III, RR 0.40). PMID- 15548193 TI - A comparative analysis of outcome after heart transplantation in patients aged 60 years and older: the University of Alberta experience. AB - BACKGROUND: The Registry of the International Society for Heart and Lung Transplantation (ISHLT) 2001 Annual Report indicated that the vast majority of heart transplant recipients are between 50 and 64 years of age. However, patient age beyond 60 years may have higher long-term mortality compared to younger patients. The purpose of this study was to compare short- and intermediate-term results including rates of acute rejection, transplant coronary artery disease, infections, malignancy, and mortality in cardiac transplant recipients 60 years or older with those below the age of 60 years. METHODS: We retrospectively analyzed the results of 50 patients aged 60 years and older who underwent heart transplantation at the University of Alberta from January 1990 to December 2000 and compared them with the results of 225 younger patients undergoing heart transplantation in the same time period. RESULTS: The older and younger groups had similar rates for treated acute rejection episodes (20.0% vs. 12.6%), transplant coronary artery disease (4.0% vs. 1.1%), and mortality (10.5% vs. 14.3%), respectively. No differences were noted with regards to quality and quantity of infection or malignancy rates. Five-year actuarial survival between the older and younger patients was also comparable at 89.5% vs.86.9% (p > 0.05). CONCLUSIONS: Heart transplantation in patients 60 years of age and older can be performed as successfully as in younger patients (< 60 years) with comparable morbidity and mortality, suggesting that patient age per se should not be an exclusion criterion for heart transplantation. PMID- 15548194 TI - The corticoid environment: a determining factor for neural progenitors' survival in the adult hippocampus. AB - New neurons continue to be generated in the adult dentate gyrus of the hippocampus. Corticosterone (CORT), a steroid secreted by the adrenal glands, had been shown to regulate progenitor proliferation. High levels of CORT suppress proliferation while low levels of the steroid stimulate it. Here we present an investigation into the regulation of survival by corticoids, with emphasis on the differential effects of the pre-mitotic and post-mitotic corticoid environments. Post-mitotic adrenalectomy increased subsequent survival of progenitors at 28 days, while additional CORT administered during the post-mitotic period decreased survival. In contrast, a corticoid-free environment prior to progenitor division resulted in a reduced survival rate of new cells and, similarly, high levels of CORT before proliferation reduced subsequent survival. In addition, phased treatment with CORT during a 27-day post-mitotic interval showed that newly formed cells lose their sensitivity to administered CORT after about 18 days. These results are the first to show that the corticoid environment both before and after cell division regulates survival. PMID- 15548195 TI - Intrinsic and spontaneous neurogenesis in the postnatal slice culture of rat hippocampus. AB - Organotypic slice culture preserves the morphological and physiological features of the hippocampus of live animals for a certain time. The hippocampus is one of exceptional regions where neurons are generated intrinsically and spontaneously throughout postnatal life. We investigated the possibility that neurons are generated continuously at the dentate granule cell layer (GCL) in slice culture of the rat hippocampus. Using 5-bromodeoxyuridine (BrdU) labelling and retrovirus vector transduction methods, the phenotypes of the newly generated cells were identified immunohistochemically. At 4 weeks after BrdU exposure, BrdU-labelled cells were found in the GCL and were immunoreactive with a neuronal marker, anti NeuN. There were fibrils immunoreactive with anti-glial fibrillary acidic protein (GFAP), an astrocyte marker, in the layer covering the GCL and occasionally encapsulated BrdU-labelled nuclei. When the newly divided cells were marked with the enhanced green fluorescent protein (EGFP) using a retrovirus vector, these cells had proliferative abilities throughout the following 4-week cultivation period. Four weeks after the inoculation, the EGFP-expressing cells consisted of various phenotypes of both early and late stages of differentiation; some were NeuN-positive cells with appearances of neurons in the GCL and some were immunoreactive with anti-Tuj1, a marker of immature neurons. Some EGFP-expressing cells were immunoreactive with anti-GFAP or anti-nestin, a marker of neural progenitors. The present study suggests that slice cultures intrinsically retain spontaneous neurogenic abilities for their cultivation period. The combination of slice culture and retrovirus transduction methods enable the newly divided cells to be followed up for a long period. PMID- 15548196 TI - BM88 is an early marker of proliferating precursor cells that will differentiate into the neuronal lineage. AB - Progression of progenitor cells towards neuronal differentiation is tightly linked with cell cycle control and the switch from proliferative to neuron generating divisions. We have previously shown that the neuronal protein BM88 drives neuroblastoma cells towards exit from the cell cycle and differentiation into a neuronal phenotype in vitro. Here, we explored the role of BM88 during neuronal birth, cell cycle exit and the initiation of differentiation in vivo. By double- and triple-labelling with the S-phase marker BrdU or the late G2 and M phase marker cyclin B1, antibodies to BM88 and markers of the neuronal or glial cell lineages, we demonstrate that in the rodent forebrain, BM88 is expressed in multipotential progenitor cells before terminal mitosis and in their neuronal progeny during the neurogenic interval, as well as in the adult. Further, we defined at E16 a cohort of proliferative progenitors that exit S phase in synchrony, and by following their fate for 24 h we show that BM88 is associated with the dynamics of neuron-generating divisions. Expression of BM88 was also evident in cycling cortical radial glial cells, which constitute the main neurogenic population in the cerebral cortex. In agreement, BM88 expression was markedly reduced and restricted to a smaller percentage of cells in the cerebral cortex of the Small eye mutant mice, which lack functional Pax6 and exhibit severe neurogenesis defects. Our data show an interesting correlation between BM88 expression and the progression of progenitor cells towards neuronal differentiation during the neurogenic interval. PMID- 15548197 TI - The extracellular matrix glycoprotein Tenascin-C is expressed by oligodendrocyte precursor cells and required for the regulation of maturation rate, survival and responsiveness to platelet-derived growth factor. AB - Analysis of Tenascin-C (TN-C) knockout mice revealed novel roles for this extracellular matrix (ECM) protein in regulation of the developmental programme of oligodendrocyte precursor cells (OPCs), their maturation into myelinating oligodendrocytes and sensitivity to growth factors. A major component of the ECM of developing nervous tissue, TN-C was expressed in zones of proliferation, migration and morphogenesis. Examination of TN-C knockout mice showed roles for TN-C in control of OPC proliferation and migration towards zones of myelination [E. Garcion et al. (2001) Development, 128, 2485-2496]. Extending our studies of TN-C effects on OPC development we found that OPCs can endogenously express TN-C protein. This expression covered the whole range of possible TN-C isoforms and could be strongly up-regulated by leukaemia inhibitory factor and ciliary neurotrophic factor, cytokines known to modulate OPC proliferation and survival. Comparative analysis of TN-C knockout OPCs with wild-type OPCs reveals an accelerated rate of maturation in the absence of TN-C, with earlier morphological differentiation and precocious expression of myelin basic protein. TN-C knockout OPCs plated on poly-lysine displayed higher levels of apoptosis than wild-type OPCs and there was also an earlier loss of responsiveness to the protective effects of platelet-derived growth factor (PDGF), indicating that TN-C has anti apoptotic effects that may be associated with PDGF signalling. The existence of mechanisms to compensate for the absence of TN-C in the knockout is indicated by the development of oligodendrocytes derived from TN-C knockout neurospheres. These were present in equivalent proportions to those found in wild-type neurospheres but displayed enhanced myelin membrane formation. PMID- 15548198 TI - Expression profiling in laser-microdissected hippocampal subregions in rat brain reveals large subregion-specific differences in expression. AB - Expression profiling in the hippocampus is hampered by its cellular heterogeneity. The aim of this study was to evaluate the feasibility of using laser-microdissected hippocampal subregions for expression profiling to improve detection of transcripts with a subregion-specific expression. Cornu ammonis (CA)3 and dentate gyrus (DG) subregions were isolated from rat brain slices using laser microdissection, subjected to two rounds of linear amplification and hybridized to rat GeneChips containing approximately 8000 transcripts (RG_U34A; Affymetrix). Analysis of the data using significance analysis of microarrays revealed 724 genes with a significant difference in expression between CA3 and DG with a false discovery rate of 2.1%, of which 264 had higher expression in DG and 460 in CA3. Several transcripts with known differential expression between the subregions were included in the dataset, as well as numerous novel mRNAs and expressed sequence tags. Sorting of the differentially expressed genes according to gene ontology classification revealed that genes involved in glycolysis and general metabolism, neurogenesis and cell adhesion were consistently expressed at higher levels in CA3. Conversely, a large cluster of genes involved in protein biosynthesis were expressed at higher levels in DG. In situ hybridization was used to validate differential expression of a selection of genes. The results of this study demonstrate that the combination of laser microdissection and GeneChip technology is both technically feasible and very promising. Besides providing an extensive inventory of genes showing differential expression between CA3 and DG, this study supports the idea that profiling in hippocampal subregions should improve detection of genes with a subregion-specific expression or regulation. PMID- 15548199 TI - Spinal cord plasticity in response to unilateral inhibition of the rat motor cortex during development: changes to gene expression, muscle afferents and the ipsilateral corticospinal projection. AB - In developing Wistar albino rats, ventral horn muscle afferent boutons are lost following corticospinal innervation. Motor cortex lesions rescue a proportion of these boutons and perturb activity dependent expression of cJun and parvalbumin (PV) in the spinal cord. Therefore, we tested whether activity-dependent competition between corticospinal and proprioreceptive afferents determines the balance of these inputs to motor output pathways by delivering the inhibitory GABA agonist muscimol unilaterally to the forelimb motor cortex using slow release polymer implants from postnatal day 7 (P7) coincident with corticospinal synaptogenesis. Controls received saline. Inhibition of immature cortical neurons by muscimol was confirmed with separate in vitro electrophysiological recordings. After P28, spinal cord sections were immunostained for PV, cJun and muscle afferents transganglionically labelled with cholera toxin-B (CTB). Unilateral inhibition reduced contralaterally the number of PV positive spinal cord neurons and muscle afferent boutons in the dorsolateral ventral horn, compared to controls, and significantly altered the distribution of motoneuronal cJun expression. Separately, descending tracts were retrogradely traced with CTB from the cervical hemicord contralateral to implants. Forelimb sensorimotor cortex sections were immunostained for either CTB or PV. In muscimol treated animals, significantly fewer neurons expressed PV in the inhibited hemicortex, but as many CTB labelled corticospinal neurons were present as in controls, along with an equally large corticospinal projection from contralateral to the implant, significantly greater than in controls. Unexpectedly, unilateral inhibition of the motor cortical input did not lead to an expanded muscle afferent input. Instead, this was reduced coincident with development of a bilateral corticospinal innervation. PMID- 15548200 TI - A soluble Nogo receptor differentially affects plasticity of spinally projecting axons. AB - In the central nervous system, regeneration of injured axons and sprouting of intact axons are suppressed by myelin-derived molecules that bind to the Nogo receptor (NgR). We used a soluble form of the NgR (sNgR), constructed as an IgG of the human NgR extracellular domain, to manipulate plasticity of uninjured primary afferent and descending monoaminergic projections to the rat spinal cord following dorsal rhizotomy. Rats with quadruple dorsal rhizotomies were treated with intrathecal sNgR or saline, or were left untreated for 2 weeks. Rhizotomy alone resulted in sprouting of serotonergic axons and to a lesser extent, tyrosine-hydroxylase (TH)-expressing axons, while axons expressing dopamine-beta hydroxylase (DbetaH) were unaffected. Human IgG immunohistochemistry revealed that sNgR infused into the intrathecal space penetrated approximately 300 microm into spinal white and grey matter. Separate axonal populations differed in their responses to intrathecal sNgR: TH-expressing and DbetaH-expressing axons responded most and least vigorously, respectively. Serotonergic axons were identified by serotonin (5-HT) or serotonin transporter (SERT) immunohistochemistry. Interestingly, a large increase in 5-HT compared to SERT positive axons density in both saline and sNgR-treated rats indicated that serotonergic axons both sprouted and increased their transmitter content in response to rhizotomy and sNgR treatment. Calcitonin gene-related peptide positive axons were largely depleted ipsilaterally by rhizotomy, and sNgR increased axon density only in deeper contralateral laminae (III-V). GAP-43 immunohistochemistry revealed a small increase in axon density following dorsal rhizotomy that was further augmented by sNgR treatment. These results reveal a differential effect of myelin antagonism on distinct populations of spinally projecting axons. PMID- 15548201 TI - Hippocampal BDNF mediates the efficacy of exercise on synaptic plasticity and cognition. AB - We found that a short exercise period enhanced cognitive function on the Morris water maze (MWM), such that exercised animals were significantly better than sedentary controls at learning and recalling the location of the platform. The finding that exercise increased brain-derived neurotrophic factor (BDNF), a molecule important for synaptic plasticity and learning and memory, impelled us to examine whether a BDNF-mediated mechanism subserves the capacity of exercise to improve hippocampal-dependent learning. A specific immunoadhesin chimera (TrkB IgG), that mimics the BDNF receptor, TrkB, to selectively bind BDNF molecules, was used to block BDNF in the hippocampus during a 1-week voluntary exercise period. After this, a 2-trial-per-day MWM was performed for 5 consecutive days, succeeded by a probe trial 2 days later. By inhibiting BDNF action we blocked the benefit of exercise on cognitive function, such that the learning and recall abilities of exercising animals receiving the BDNF blocker were reduced to sedentary control levels. Inhibiting BDNF action also blocked the effect of exercise on downstream systems regulated by BDNF and important for synaptic plasticity, cAMP response-element-binding protein (CREB) and synapsin I. Specific to exercise, we found an association between CREB and BDNF expression and cognitive function, such that animals who were the fastest learners and had the best recall showed the highest expression of BDNF and associated CREB mRNA levels. These findings suggest a functional role for CREB under the control of BDNF in mediating the exercise-induced enhancement in learning and memory. Our results indicate that synapsin I might also contribute to this BDNF-mediated mechanism. PMID- 15548202 TI - Identified spinal motoneurons of young rats possess nicotinic acetylcholine receptors of the heteromeric family. AB - The aim of the present study was to determine whether, in young rats, spinal motoneurons possess functional nicotinic acetylcholine receptors. Motoneurons were identified either by retrograde labelling or by choline acetyltransferase immunohistochemistry. Whole-cell recordings were performed in spinal cord slices cut at the lumbar level. In voltage clamp, acetylcholine evoked a rapidly activating inward current. In current clamp, it depolarized the motoneuron membrane and induced action potential firing. The acetylcholine-evoked current was strongly reduced by d-tubocurarine or dihydro-beta-erythroidine, broad spectrum nicotinic antagonists, but was almost insensitive to methyllycaconitine, a nicotinic antagonist selective for receptors containing the alpha7 subunit. Moreover, exo-2-(2-pyridyl)-7-azabicyclo[2.2.1]heptane, an alpha7-specific agonist, was without effect. In young animals, light-microscopic autoradiography showed that in the central grey matter all laminae were intensely and equally labelled by [3H]epibatidine. A dense [125I]-alpha-bungarotoxin binding was also found in all laminae, with slightly lower levels in the superficial layers of the dorsal horns and in the ventral part of the grey matter. In adults, the density of [3H]epibatidine binding sites was much lower in the entire grey matter, except in layer 2 of the dorsal horn, and [125I]-alpha-bungarotoxin binding sites were present only in some selected areas. Our data indicate that spinal motoneurons possess functional nicotinic receptors of the heteromeric type and suggest that nicotinic cholinergic transmission may play a significant role in the developing spinal cord. PMID- 15548203 TI - Putative NMDA receptors in Hydra: a biochemical and functional study. AB - The feeding behaviour of the freshwater polyp Hydra vulgaris (Cnidaria, Hydrozoa) is modulated by a number of molecules acting as neurotransmitters in other nervous systems. Here we present biochemical and functional evidence of the occurrence of putative NMDA receptors in Hydra tissues. Saturation experiments showed the presence of one population of binding sites with nanomolar affinity and low capacity for [3H]MK-801. Before equilibrium, [3H]MK-801 binding was increased by the agonists glutamate and glycine as well as by reduced glutathione (GSH). In vivo the glutamate receptor agonist NMDA markedly decreased the duration of the response to GSH. This effect was linearly related to ligand doses in the nanomolar concentration range and was counteracted by either the NMDAR specific antagonist D-AP5 or by the d-serine antagonist DCKA. When NMDA concentration was increased to 10 or 100 microm, duration of the response to GSH was no longer affected unless the lectin concanavalin A, which prevents receptor desensitization in other systems, was added to the test medium. Simultaneous administration of ineffective doses of NMDA and strychnine, glycine or d-serine, an agonist at the glycine binding site of the NMDA receptor in vertebrate CNS, resulted in a strong reduction of response duration. Both D-AP5 and DCKA suppressed this effect. These results, together with the decrease in response duration produced by d-serine, support the hypothesis that NMDA-like glutamate receptors may occur in Hydra tissues where they are involved in modulation of the response to GSH with opposite actions to those of GABA and glycine. PMID- 15548204 TI - Cellular prion protein/laminin receptor: distribution in adult central nervous system and characterization of an isoform associated with a subtype of cortical neurons. AB - The 67-kDa LR protein was originally discovered as a non-integrin laminin receptor. Several more recent in vitro studies demonstrated the function of 67 kDa LR and its related 'precursor' form 37-kDa LRP as receptors of cellular prion protein and their implication in abnormal prion protein propagation in vitro. In addition, expression of both proteins was shown to increase considerably in the brain of scrapie-infected mice and hamsters. While LRP/LR are thus likely to play important roles in neuronal cell adhesion, survival and homeostasis and during pathological disorders, little is known so far about their fine cellular distribution in adult central nervous system. Using immunocytochemistry and western blotting, we show here that the 67-kDa LR is the major receptor form in adult rat brain and spinal cord, expressed within the cytoplasm and at the plasma membrane of most neurons and in a subset of glial cells. The overall distribution of LR correlates well with that reported for laminin-1 but also with brain regions classically associated with prion-related neurodegeneration. In contrast to LR, the 37-kDa LRP form is much less abundant in adult than in postnatal central nervous system. Characterization of a novel antibody allowed us to study the distribution across tissues of cell membrane-associated LRP. Interestingly, this form is almost exclusively found on a subclass of parvalbumin-immunoreactive cortical interneurons known to degenerate during the early stages of Creutzfeldt Jakob disease. Our demonstration of local differences in the expression of particular LRP/LR isoforms may be a first step towards unraveling their specific molecular interactions. PMID- 15548205 TI - Distribution and signaling of TREM2/DAP12, the receptor system mutated in human polycystic lipomembraneous osteodysplasia with sclerosing leukoencephalopathy dementia. AB - Together with its adaptor protein, the adaptor protein of 12 kDa also known as KARAP and TYROBP (DAP12), triggering receptor expressed in myeloid cells 2 (TREM2) is a stimulatory membrane receptor of the immunoglobulin/lectin-like superfamily, well known in myeloid cells. In humans, however, loss-of-function mutations of TREM2/DAP12 leave myeloid cells unaffected but induce an autosomal recessive disease characterized, together with bone cysts, by a spectrum of pathological lesions in the cortex, thalamus and basal ganglia with clinical symptoms of progressive dementia (polycystic lipomembraneous osteodysplasia with sclerosing leukoencephalopathy). Nothing was known about the role of TREM2/DAP12 in brain cell biology and physiology. By confocal immunocytochemistry we demonstrate that, in both human and mouse cerebral cortex, TREM2/DAP12, strongly expressed by microglia, is also present in a fraction of neurons but not in astrocytes and oligodendrocytes. In contrast, in the hippocampal cortex TREM2 expressing neurons are rare. Both in neurons and microglia the receptor appears to be located mostly intracellularly in a discrete compartment(s) partially coinciding with (or adjacent to) the Golgi complex/trans-Golgi network. Four nerve cell lines were identified as expressing the intracellular receptor system. In living human microglia CHME-5 and glioblastoma T98G cells, activation of TREM2 by its specific antibody induced [Ca2+]i responses, documenting its surface expression and functioning. Surface expression of TREM2, low in resting CHME-5 and T98G cells, increases significantly and transiently (60 min) when cells are stimulated by ionomycin, as revealed by both surface biotinylation and surface immunolabeling. Our results provide the first information about the expression, distribution (mostly intracellular) and functioning of TREM2/DAP12 system in nerve cells, a necessary step in the understanding of the cellular mechanisms affected in polycystic lipomembraneous osteodysplasia with sclerosing leukoencephalopathy. PMID- 15548206 TI - Involvement of corticostriatal glutamatergic terminals in striatal dopamine release elicited by stimulation of delta-opioid receptors. AB - We have previously shown that striatal dopamine release induced locally by a delta-opioid receptor agonist was totally inhibited by a glutamate N-methyl-D aspartate receptor antagonist, indicating the involvement of glutamatergic receptors in this effect. The aim of the present study was to specify this mechanism. Firstly, we investigated the effect of [D-Pen2,D-Pen5]-enkephalin (DPDPE) on glutamate release in rats by intrastriatal microdialysis. The infusion of DPDPE (10 microm) enhanced the glutamate content in dialysate by approximately 34%, an effect which did not appear to result from inhibition of glutamate uptake. We then considered the consequences of a unilateral thermocoagulation of the frontal cortex on either glutamate or dopamine release induced by stimulation of delta-opioid receptors 2 days later. This lesion, which decreased the glutamate content in ipsilateral striatum by approximately 30%, totally prevented the increase in dialysate levels of glutamate induced by DPDPE. Moreover, whereas DPDPE (10 microm) was found to increase the striatal dopamine release in intact animals by approximately 59%, this effect was also completely suppressed by the cortical lesion. Finally, we studied the effect of the lesion on the [3H]-DPDPE binding to striatal membranes prepared from the whole striatum. In the ipsilateral striatum a significant decrease in this [3H]-DPDPE binding (by approximately 18%) was found 2 days after the lesion. Our results indicate that the increase in striatal dopamine release induced by DPDPE probably depends on glutamate release from corticostriatal glutamatergic afferents in response to the stimulation of delta-opioid receptors located on terminals of these neurons. PMID- 15548207 TI - Delayed onset of prepulse inhibition deficits following kainic acid treatment on postnatal day 7 in rats. AB - Abnormal activity in corticolimbic circuits during development may be a predisposing factor for schizophrenia. Permanent or temporary lesions of limbic structures such as the ventral hippocampus and basolateral amygdala in rats on postnatal day (PND) 7 result in functional changes similar to some behavioural and cognitive signs of schizophrenia. The present experiments tested whether transient increases in the neural activity of corticolimbic circuits on PND 7 would result in similar behavioural changes. Long-Evans rats were treated with either kainic acid (KA, 1.5 mg/kg, i.p.) or saline on PND 7 and tested for prepulse inhibition (PPI) of the acoustic startle response and spontaneous locomotor activity both in a novel environment and following amphetamine treatment before puberty (PND 35) and in early adulthood (PND 56). In subgroups of animals PPI was also measured following apomorphine administration (0.2 mg/kg) and spatial learning and memory were tested in the water maze. Rats treated with KA were indistinguishable from saline-treated animals on PND 35. However, on PND 56, KA-treated animals showed a subtle consistent decrease in PPI relative to control animals, but did not show increased sensitivity to the disruptive effects of a low dose of apomorphine on PPI. Locomotor responses to novelty or amphetamine were not reliably altered in the KA-treated animals. KA- and saline treated animals performed similarly in the water maze. These results support the hypothesis that neural hyperactivity on PND 7 in rats causes behavioural changes in early adulthood that resemble some symptoms of schizophrenia. These pharmacological data suggest that the changes are not mediated by postsynaptic alterations in mesolimbic dopamine transmission. PMID- 15548208 TI - Dynamic regulation of spine-dendrite coupling in cultured hippocampal neurons. AB - We investigated the role of dendritic spine morphology in spine-dendrite calcium communication using novel experimental and theoretical approaches. A transient rise in [Ca2+]i was produced in individual spine heads of Fluo-4-loaded cultured hippocampal neurons by flash photolysis of caged calcium. Following flash photolysis in the spine head, a delayed [Ca2+]i transient was detected in the parent dendrites of only short, but not long, spines. Delayed elevated fluorescence in the dendrite of the short spines was also seen with a membrane bound fluorophore and fluorescence recovery from bleaching of a calcium-bound fluorophore had a much slower kinetics, indicating that the dendritic fluorescence change reflects a genuine diffusion of free [Ca2+]i from the spine head to the parent dendrite. Calcium diffusion between spine head and the parent dendrite was regulated by calcium stores as well as by a Na-Ca exchanger. Spine length varied with the recent history of the [Ca2+]i variations in the spine, such that small numbers of calcium transients resulted in elongation of spines whereas large numbers of calcium transients caused shrinkage of the spines. Consequently, spine elongation resulted in a complete isolation of the spine from the dendrite, while shrinkage caused an enhanced coupling with the parent dendrite. These studies highlight a dynamically regulated coupling between a dendritic spine head and its parent dendrite. PMID- 15548209 TI - Optical imaging of the spatiotemporal dynamics of cerebral blood flow and oxidative metabolism in the rat barrel cortex. AB - Oxidative metabolism and cerebral blood flow (CBF) are two of the most important measures in neuroimaging. However, results from concurrent imaging of the two with high spatial and temporal resolution have never been published. We used flavoprotein autofluorescence (AF) and laser speckle imaging (LSI) in the anaesthetized rat to map oxidative metabolism and CBF in response to single vibrissa stimulation. Autofluorescence responses reflecting oxidative metabolism demonstrated a fast increase with a delay of 0.1 s. The sign-reversed speckle contrast reflecting CBF started to rise with a delay of 0.6 s and reached its maximum 1.4 s after the stimulation offset. The fractional signal changes were 2.0% in AF and 9.7% in LSI. Pixelwise modelling revealed that CBF maps spread over an area up to 2.5-times larger than metabolic maps. The results provide evidence that the increase in cerebral oxidative metabolism in response to sensory stimulation is considerably faster and more localized than the CBF response. This suggests that future developments in functional imaging concentrating on the metabolic response promise an increased spatial resolution. PMID- 15548210 TI - A role for ERKII in synaptic pattern selectivity on the time-scale of minutes. AB - Stimulus reinforcement strengthens learning. Intervals between reinforcement affect both the kind of learning that occurs and the amount of learning. Stimuli spaced by a few minutes result in more effective learning than when massed together. There are several synaptic correlates of repeated stimuli, such as different kinds of plasticity and the amplitude of synaptic change. Here we study the role of signalling pathways in the synapse on this selectivity for spaced stimuli. Using the in vitro hippocampal slice technique we monitored long-term potentiation (LTP) amplitude in CA1 for repeated 100-Hz, 1-s tetani. We observe the highest LTP levels when the inter-tetanus interval is 5-10 min. We tested biochemical activity in the slice following the same stimuli, and found that extracellular signal-regulated kinase type II (ERKII) but not CaMKII exhibits a peak at about 10 min. When calcium influx into the slice is buffered using AM ester calcium dyes, amplitude of the physiological and biochemical response is reduced, but the timing is not shifted. We have previously used computer simulations of synaptic signalling to predict such temporal tuning from signalling pathways. In the current study we consider feedback and feedforward models that exhibit temporal tuning consistent with our experiments. We find that a model incorporating post-stimulus build-up of PKM zeta acting upstream of mitogen-activated protein kinase is sufficient to explain the observed temporal tuning. On the basis of these combined experimental and modelling results we propose that the dynamics of PKM activation and ERKII signalling may provide a mechanism for functionally important forms of synaptic pattern selectivity. PMID- 15548211 TI - Input and frequency-specific entrainment of postsynaptic firing by IPSPs of perisomatic or dendritic origin. AB - Correlated activity of cortical neurons underlies cognitive processes. Networks of several distinct classes of gamma-aminobutyric acid (GABA)ergic interneurons are capable of synchronizing cortical neurons at behaviourally relevant frequencies. Here we show that perisomatic and dendritic GABAergic inputs provided by two classes of GABAergic cells, fast spiking and bitufted interneurons, respectively, entrain the timing of postsynaptic spikes differentially in both pyramidal cells and interneurons at beta and gamma frequencies. Entrainment of pyramidal as well as regular spiking non-pyramidal cells was input site and inhibitory postsynaptic potential frequency dependent. Gamma frequency input from fast spiking cells entrained pyramidal cells on the positive phase of an intrinsic cellular theta oscillation, whereas input from bitufted cells was most effective in gamma frequency entrainment on the negative phase of the theta oscillation. The discharge of regular spiking interneurons was phased at gamma frequency by dendritic input from bitufted cells, but not by perisomatic input from fast spiking cells. Action potentials in fast spiking GABAergic neurons were phased at gamma frequency by both other fast spiking and bitufted cells, regardless of whether the presynaptic GABAergic input was at gamma or beta frequency. The interaction of cell type-specific intrinsic properties and location-selective GABAergic inputs could result in a spatio temporally regulated synchronization and gating of cortical spike propagation in the network. PMID- 15548212 TI - The cortically evoked secondary depolarization affects the integrative properties of thalamic reticular neurons. AB - Corticothalamic terminals on thalamic reticular (RE) neurons account for most synapses from afferent pathways onto this nucleus and these inputs are more powerful than those from axon collaterals of thalamocortical neurons. Given the supremacy of cortical inputs, we analysed here the characteristics and possible mechanisms underlying a secondary component of the cortically elicited depolarization in RE neurons, recorded in cats under barbiturate anesthesia. Electrical stimulation of corticothalamic axons in the internal capsule evoked fixed and short-latency excitatory postsynaptic potentials (EPSPs) that, by increasing stimulation intensity and at hyperpolarized levels (< -70 mV), developed into low-threshold spikes and spindle oscillations. The threshold for spindle oscillations was 60% higher than that required for evoking minimal EPSPs. The evoked EPSPs included a secondary depolarizing component, which appeared approximately 5 ms after the peak of the initial component and was voltage dependent, i.e. most prominent between -70 mV and -85 mV, while being greatly reduced or absent at more hyperpolarized levels. The secondary depolarizing component was sensitive to QX-314 in the recording micropipette. We suggest that the secondary component of cortically evoked EPSPs in RE neurons is due to the dendritic activation of T-currents, with a probable contribution of the persistent Na+ current. This late component affected the integrative properties of RE neurons, including their spiking output and temporal summation of incoming cortical inputs. PMID- 15548213 TI - Interneurone bursts are spontaneously associated with muscle contractions only during early phases of mouse spinal network development: a study in organotypic cultures. AB - For a short time during development immature circuits in the spinal cord and other parts of the central nervous system spontaneously generate synchronous patterns of rhythmic activity. In the case of the spinal cord, it is still unclear how strongly synchronized bursts generated by interneurones are associated with motoneurone firing and whether the progressive decline in spontaneous bursting during circuit maturation proceeds in parallel for motoneurone and interneurone networks. We used organotypic cocultures of spinal cord and skeletal muscle in order to investigate the ontogenic evolution of endogenous spinal network activity associated with the generation of coordinate muscle fibre contractions. A combination of multiunit electrophysiological recordings, videomicroscopy and optical flow computation allowed us to measure the correlation between interneurone firing and motoneurone outputs after 1, 2 and 3 weeks of in vitro development. We found that, in spinal organotypic slices, there is a developmental switch of spontaneous activity from stable bursting to random patterns after the first week in culture. Conversely, bursting recorded in the presence of strychnine and bicuculline became increasingly regular with time in vitro. The time course of spontaneous activity maturation in organotypic slices is similar to that previously reported for the spinal cord developing in utero. We also demonstrated that spontaneous bursts of interneurone action potentials strongly correlate with muscular contractions only during the first week in vitro and that this is due to the activation of motoneurones via AMPA type glutamate receptors. These results indicate the occurrence in vitro of motor network development regulating bursting inputs from interneurones to motoneurones. PMID- 15548214 TI - Somatostatin receptors differentially affect spontaneous epileptiform activity in mouse hippocampal slices. AB - Somatostatin-14 [somatotropin release-inhibiting factor (SRIF)] reduces hippocampal epileptiform activity but the contribution of its specific receptors (sst1-5) is poorly understood. We have focused on the role of sst1 and sst2 in mediating SRIF modulation of epilepsy using hippocampal slices of wild-type (WT) and sst1 or sst2 knockout (KO) mice. Recordings of epileptiform discharge induced by Mg2+ -free medium with 4-aminopyridine were performed from the CA3 region before and after the application of SRIF compounds. In WT mice, SRIF and the sst1 agonist CH-275 reduce epilepsy whereas sst1 blockade with its antagonist SRA-880 increases the bursting discharge. Activation of sst2 does not affect the bursting frequency unless its agonist octreotide is applied with SRA-880, indicating that sst1 masks sst2-mediated modulation of epilepsy. In sst1 KO mice: (i) the bursting frequency is lower than in WT; (ii) SRIF, CH-275 and SRA-880 are ineffective on epilepsy and (iii) octreotide is also devoid of effects, whereas blockade of sst2 with the antagonist D-Tyr8 Cyn 154806 increases the bursting frequency. In sst2 KO mice, the SRIF ligand effects are similar to those in WT. In the whole hippocampus of sst1 KO mice, sst2 mRNA, protein and binding are higher than in WT and reverse transcription-polymerase chain reaction of the CA3 subarea confirms an increase of the sst2 messenger. We conclude that sst1 mediates inhibitory actions of SRIF and that interactions between sst1 and sst2 may prevent sst2 modulation of epilepsy. We suggest that, in sst1 KO mice, activation of over-expressed sst2 reduces the bursting frequency, indicating that sst2 density represents the rate-limiting factor for ss(2-mediated modulation of epilepsy. PMID- 15548215 TI - Functional neuroanatomy of the human near/far response to blur cues: eye-lens accommodation/vergence to point targets varying in depth. AB - The purpose of this study was to identify the networks involved in the regulation of visual accommodation/vergence by contrasting the cortical functions subservient to eye-lens accommodation with those evoked by foveal fixation. Neural activity was assessed in normal volunteers by changes in rCBF measured with PET. Thirteen right-handed subjects participated in three monocular tasks: (i) resting with eyes closed; (ii) sustained foveal fixation upon a LED at 1.2 m (0.83 D); and (iii) accommodating alternately on a near (24 cm, 4.16 D) vs. a far (3.0 m, 0.33 D) LED alternately illuminated in sequential 2 s epochs. The contrast between the conditions of near/far accommodation and of constant foveal fixation revealed activation in cerebellar hemispheres and vermis; middle and inferior temporal cortex (BA 20, 21, 37); striate cortex and associative visual areas (BA 17/18). Comparison of the condition of constant fixation with the condition of resting with closed eyes indicated activation of cerebellar hemispheres and vermis; visual cortices (BA 17/18); a right hemisphere dominant network encompassing prefrontal (BA 6, 9, 47), superior parietal (BA 7), and superior temporal (BA 40) cortices; and bilateral thalamus. The contrast between the conditions of near/far accommodation with closed-eye rest reflected an incremental summation of the activations found in the previous comparisons (i.e. activations associated with constant fixation). Neural circuits activated selectively during the near/far response to blur cues over those during constant visual fixation, occupy posterior structures that include occipital visual regions, cerebellar hemispheres and vermis, and temporal cortex. PMID- 15548216 TI - Acute improvement of hand sensibility after selective ipsilateral cutaneous forearm anaesthesia. AB - The cortical representation of body parts is constantly modulated in response to the afferent input, and acute deafferentation of a body part results in bilateral cortical reorganization. To study the effects on hand function of right forearm anaesthesia, we investigated ten human subjects (group 1) for perception of touch, tactile discrimination and grip strength in the right (ipsilateral) and left (contralateral) hand before, during and 24 h after forearm skin anaesthesia with a local anaesthetic cream (EMLA). Ten age-matched controls (group 2) were investigated in the same way but received placebo. In group 1 a significant improvement was seen in tactile discrimination in the ipsilateral hand compared to base line (P = 0.009) and compared to group 2 (P = 0.006). The improvement in tactile discrimination remained for at least 24 h after anaesthesia. Perception of touch, was improved during anaesthesia compared to baseline values in group 1 (P = 0.046) and remained for at least 24 h. Grip strength did not change. These findings suggest that transient selective deafferentation of an extremity results in enhanced sensory functions of the functionally preserved parts of the same extremity, presumably as a result of expansion of adjacent cortical territories. Such rapid functional changes suggest unmasking of pre-existing synaptic connections as the mechanism underlying the acute modulation of sensory functions in the hand. Our findings open new perspectives for sensory re-education and rehabilitation following injury to the peripheral and central nervous system. PMID- 15548217 TI - Delta9-tetrahydrocannabinol decreases somatic and motivational manifestations of nicotine withdrawal in mice. AB - The possible interactions between Delta9-tetrahydrocannabinol (Delta9-THC) and nicotine remain unclear in spite of the current association of cannabis and tobacco in humans. The aim of the present study was to explore the interactions between these two drugs of abuse by evaluating the consequences of Delta9-THC administration on the somatic manifestations and the aversive motivational state associated with nicotine withdrawal in mice. Acute Delta9-THC administration significantly decreased the incidence of several nicotine withdrawal signs precipitated by mecamylamine or naloxone, such as wet-dog-shakes, paw tremor and scratches. In both experimental conditions, the global withdrawal score was also significantly attenuated by acute Delta9-THC administration. This effect of Delta9-THC was not due to possible adaptive changes induced by chronic nicotine on CB1 cannabinoid receptors, as the density and functional activity of these receptors were not modified by chronic nicotine administration in the different brain structures investigated. We also evaluated the consequences of Delta9-THC administration on c-Fos expression in several brain structures after chronic nicotine administration and withdrawal. c-Fos was decreased in the caudate putamen and the dentate gyrus after mecamylamine precipitated nicotine withdrawal. However, acute Delta9-THC administration did not modify c-Fos expression under these experimental conditions. Finally, Delta9-THC also reversed conditioned place aversion associated to naloxone precipitated nicotine withdrawal. Taken together, these results indicate that Delta9-THC administration attenuated somatic signs of nicotine withdrawal and this effect was not associated with compensatory changes on CB1 cannabinoid receptors during chronic nicotine administration. In addition, Delta9-THC also ameliorated the aversive motivational consequences of nicotine withdrawal. PMID- 15548218 TI - Altered morphological and electrophysiological properties of Cajal-Retzius cells in cerebral cortex of embryonic Presenilin-1 knockout mice. AB - Mutations of Presenilin-1 are the major cause of familial Alzheimer's disease. Presenilin-1 knockout (PS1-/-) mice develop severe cortical dysplasia related to human type 2 lissencephaly. This overmigration syndrome has been attributed to the premature loss of Cajal-Retzius cells (CRcs), pioneer neurons required for the termination of radial neuronal migration. To elucidate the potential cellular mechanisms responsible for this premature neuronal loss, we investigated the morphological and electrophysiological properties of visually identified CRcs of wild-type (WT) and PS1-/- mouse brains at embryonic day 16.5. The density of CRcs was substantially reduced in the cerebral cortex of PS1-/-. In PS1-/- CRcs the number of axonal branches was significantly increased to 12.5 +/- 4.9 (n = 8; WT, 4.0 +/- 1.4, n = 12), while no differences in dendritic branching and total length of dendritic and axonal compartments were observed. Additionally, the resting membrane potential of PS1-/- CRcs was significantly depolarized (-48.3 +/ 1.7 mV; n = 23) in contrast to WT CRcs (-57.9 +/- 2.1 mV; n = 38). Active membrane properties were not affected by PS1 deficiency. CRcs of both genotypes showed spontaneous postsynaptic currents that could be completely blocked by 100 microM bicuculline and were unaffected by glutamatergic antagonists, suggesting that they were mediated by GABAA receptors. These results demonstrate that axonal branching and resting membrane potential of CRcs was affected in embryonic cerebral cortex of PS1-/- mice. The depolarized membrane potential observed in PS1-/- CRcs may increase the susceptibility to neuronal death, thus facilitating the premature loss of CRcs in PS1-/- mice. PMID- 15548219 TI - Altered circadian locomotor activity in APP23 mice: a model for BPSD disturbances. AB - Over the past decade, clinical Alzheimer's disease research has been challenged with an increased interest in noncognitive symptomatology, commonly referred to as behavioural and psychological signs and symptoms of dementia (BPSD). In accordance, major attention is being paid to behavioural alterations in the phenotyping of transgenic mouse models. Besides an age-dependent decline of cognitive functions, the APP23 model was previously shown to exhibit cage activity disturbances, reminiscent of diurnal rhythm disturbances in Alzheimer patients. To further scrutinize these observations, circadian patterns of horizontal locomotor activity were assessed in 3-, 6- and 12-month-old APP23 mice and wild-type littermates in a test paradigm continuously recording cage activity over a period ranging from 1 to 3 days. At the age of 3 months, APP23 profiles resembled the wild-type pattern to a large extent, although minor differences were already noticeable. Six-month-old APP23 mice displayed an altered activity profile with a first indication of increased activity during the second half of the active phase, reminiscent of sundowning behaviour in Alzheimer patients. This bimodal overnight activity pattern became even more evident at the age of 12 months. The APP23 model was therefore shown to display an age-dependent development of cage activity disturbances and sundowning-like behaviour. A comparison is made with actigraphic recordings of human Alzheimer patients exhibiting sundowning behaviour. This first report of diurnal rhythm disturbances and sundowning-like phenomena in a transgenic mouse model greatly adds to the validity of the APP23 model. PMID- 15548220 TI - Direct visual and circadian pathways target neuroendocrine cells in primates. AB - The effect of light on neuroendocrine functions is thought to be mediated through retinal inputs to the circadian pacemaker, the hypothalamic suprachiasmatic nucleus (SCN). The present studies were conducted to provide experimental evidence for this signaling modality in non-human primates. In the St. Kitts vervet monkey, anterograde tracing of SCN efferents revealed a monosynaptic pathway between the circadian clock and hypothalamic neurons producing luteinizing hormone-releasing hormone (LHRH). Using a variety of tracing techniques, direct retinal input was found to be abundant in the SCN and in other hypothalamic sites. Strikingly, in hypothalamic areas other than the SCN, primary visual afferents established direct contacts with neuroendocrine cells including those producing LHRH and dopamine, neurons that are the hypothalamic regulators of pituitary gonadotrops and prolactin. Thus, our data reveal for the first time in primates that light stimuli can reach the hypothalamo-pituitary-gonadal axis, directly providing a pathway independent of but parallel to that of the circadian clock for the photic modulation of hormone release. PMID- 15548221 TI - Nucleus accumbens neurons in the rat exhibit differential activity to conditioned reinforcers and primary reinforcers within a second-order schedule of saccharin reinforcement. AB - The nucleus accumbens has been associated with processing information related to primary reinforcement and reward. Most neurophysiological studies report that nucleus accumbens neurons are phasically excited in response to the onsets of salient events during the seeking of reinforcement and to the delivery of primary reinforcers. However, a minority of studies report inhibition during primary reinforcement. We recorded from 65 neurons in the nucleus accumbens whilst thirsty rats performed under a second-order schedule of saccharin reinforcement. This allowed us to analyse neural activity and behaviour during reinforcer seeking in the presence of conditioned reinforcers (second-order stimuli, also called 'conditioned stimuli'), and during primary reinforcer consumption. Specifically, we sought to examine the valence of potential neural responses to primary reinforcement, to compare these responses to second-order stimulus-evoked responses, and to determine whether responses were differential to second-order stimuli presented at different time points within the schedule. Fifty out of 65 neurons we sampled responded to the second-order stimulus and/or consumption of the primary reinforcer. Most neurons in our sample exhibited excitation following the second-order stimulus and inhibition to the primary reinforcer, a pattern also present over the average response of the neural population. However, there was no systematic variation in neural responses evoked by second-order stimuli presented at different temporal proximities to primary reinforcement. Our results provide evidence that partially overlapping mechanisms within the nucleus accumbens differentially process conditioned reinforcers and primary reinforcers. PMID- 15548222 TI - Modular biased-competition and cooperation: a candidate mechanism for selective working memory. AB - Prefrontal cortex (PFC) has been suggested to play an important role in executive cognitive functions, participating in planning and controlling behaviour. The results of several recent electrophysiological studies indicate that PFC might be involved not only in the active maintenance of information but in doing so in a context- or task-dependent manner. In a delayed-match-to-sample paradigm, recordings from neurons in the PFC showed their ability to selectively represent information, which is needed for task completion, suggesting that task-irrelevant information does not access working memory. We present a neurodynamical computational model of a part of the PFC to account for the selective representation of information in working memory. We show that a network of biological realistic integrate-and-fire excitatory and inhibitory neurons, implementing the mechanisms of local or modular biased-competition, which is transmitted through cooperation to different subsets of neuronal pools, can explain the formation of selective context-dependent working memory. The modes of operation of the network are characterized and the corresponding parameter settings revealed. Modular competition and cooperation might constitute general mechanisms for implementing context-dependent formation of working memory. PMID- 15548223 TI - Post-training intrabasolateral amygdala infusions of dopamine modulate consolidation of inhibitory avoidance memory: involvement of noradrenergic and cholinergic systems. AB - There is extensive evidence that several neurotransmitter systems within the basolateral amygdala (BLA) influence memory consolidation. The present study investigated the influence of dopamine (DA) in the BLA on the consolidation of memory for inhibitory avoidance (IA) training. Male Sprague-Dawley rats (approximately 300 g) were trained on a step-through IA task and, 48 h later, tested for retention as indexed by their latencies to enter the shock compartment on the test day. Drugs were infused into the BLA or central amygdala nucleus (CEA) immediately or 3 h after training via bilateral cannulae. DA infused into the BLA immediately after training enhanced retention, whereas DA infused into the BLA 3 h after training or into the CEA did not affect retention. Infusions of the dopaminergic antagonist cis-Flupenthixol together with DA blocked the DA induced memory enhancement. Immediate post-training intra-BLA infusions of the D1 receptor antagonist SCH 23390 or the D2 receptor antagonist sulpiride impaired retention. beta-adrenergic or muscarinic cholinergic receptor antagonists coinfused into the BLA with DA blocked the memory enhancing effects of DA. These findings indicate that dopaminergic activation within the BLA modulates memory consolidation and that the modulation involves activation of both D1 and D2 receptors and concurrent activation of beta-adrenergic and cholinergic influences within the BLA. PMID- 15548224 TI - Are switches in perception of the Necker cube related to eye position? AB - The issue of the relation of eye position to perceptual reversals of the ambiguous figure of the 'Necker cube' dates back to Necker's original article [L.A. Necker (1832) The London & Edinburgh Philosophical Magazine and Journal of Science, 1, 329-337]. Despite the investigations of many distinguished psychophysicists since then, the question of whether perceptual switching is a cause or a consequence of associated changes in eye position has remained a matter of debate. In the present study we overcame methodological problems that have bedevilled many previous studies. We avoided any instruction that could interfere with the human subjects' free viewing of the Necker cube, tracked the eye position precisely and used biased versions of the cube that produced unambiguous percepts to determine how each subject actually looked at the cube. We show that, under these free-viewing conditions, there is a close link between the perception of the Necker cube and eye position. The average eye position of most subjects is at an extreme value at about the time when the subject's perception switches. From the biased cube trials we can infer that the polarity of the extreme corresponds to the percept which the subject had before the switch. These data indicate a bidirectional coupling between eye position and perceptual switching so that, after a subject's perceptual state changes, their eye position shifts to view the newly established percept. When the eye position approaches the corresponding extreme, the percept, in turn, becomes more and more likely to switch. This result suggests that the changed eye position itself might provide a negative feedback signal that suppresses the percept. PMID- 15548225 TI - Newborn human brain identifies repeated auditory feature conjunctions of low sequential probability. AB - Natural environments are usually composed of multiple sources for sounds. The sounds might physically differ from one another only as feature conjunctions, and several of them might occur repeatedly in the short term. Nevertheless, the detection of rare sounds requires the identification of the repeated ones. Adults have some limited ability to effortlessly identify repeated sounds in such acoustically complex environments, but the developmental onset of this finite ability is unknown. Sleeping newborn infants were presented with a repeated tone carrying six frequent (P = 0.15 each) and six rare (P approximately 0.017 each) conjunctions of its frequency, intensity and duration. Event-related potentials recorded from the infants' scalp were found to shift in amplitude towards positive polarity selectively in response to rare conjunctions. This finding suggests that humans are relatively hard-wired to preattentively identify repeated auditory feature conjunctions even when such conjunctions occur rarely among other similar ones. PMID- 15548226 TI - Altered sensorimotor development in a transgenic mouse model of amyotrophic lateral sclerosis. AB - Most neurodegenerative diseases become manifest at an adult age but abnormalities or pathological symptoms appear earlier. It is important to identify the initial mechanisms underlying such progressive neurodegenerative disease in both humans and animals. Transgenic mice expressing the familial amyotrophic lateral sclerosis (ALS)-linked mutation (G85R) in the enzyme superoxide dismutase 1 (SOD1) develop motor neuron disease at 8-10 months of age. We address the question of whether the mutation has an early impact on spinal motor networks in postnatal mutant mice. Behavioural tests showed a significant delay in righting and hind-paw grasping responses in mutant SOD1G85R mice during the first postnatal week, suggesting a transient motor deficit compared to wild-type mice. In addition, extracellular recordings from spinal ventral roots in an in vitro brainstem-spinal cord preparation demonstrated different pharmacologically induced motor activities between the two strains. Rhythmic motor activity was difficult to evoke with N-methyl-DL-aspartate and serotonin at the lumbar levels in SOD1G85R mice. In contrast to lumbar segments, rhythmic activity was similar in the sacral roots from the two strains. These results strongly support the fact that the G85R mutation may have altered lumbar spinal motor systems much earlier than previously recognized. PMID- 15548227 TI - Reelin receptors in developing laminated brain structures of mouse and human. AB - Reelin is an extracellular matrix protein secreted by a variety of cell types throughout the developing brain. The target cells for reelin express the cytoplasmic adapter protein Dab1, which binds to the reelin receptors VLDLR and ApoER2. In the present work, we have studied the localization of both receptors in developing mouse and human cortex, olfactory bulb and cerebellum. In mouse, some Cajal-Retzius cells express reelin and VLDLR; in humans, all the components of the signalling pathway (Reelin, Dab1, VLDLR and ApoER2) are present in subsets of Cajal-Retzius cells. In the mouse cortical plate, VLDLR and ApoER2 are present from E15 to postnatal stages; in human cortical plate they are most prominent at approximately 20 gestational weeks. In mice, cerebellar Purkinje cells only express VLDLR whereas in humans they express both VLDLR and ApoER2. Mitral cells of the mouse olfactory bulb are ApoER2-positive and VLDLR-negative. In sum, the receptor expression patterns are similar in the human and mouse cortical plate but differ in Cajal-Retzius and Purkinje cells, which in humans express additional components of the reelin-Dab1 pathway. PMID- 15548228 TI - A single high dose of cocaine induces behavioural sensitization and modifies mRNA encoding GluR1 and GAP-43 in rats. AB - Neuroadaptive changes underlying repeated exposure to cocaine-induced behavioural sensitization have been related to modification in the pattern of synaptic connectivity and excitatory transmission. Remarkably, even a single exposure to abused drugs is sufficient to elicit lasting behavioural sensitization. The present study investigated whether in Sprague-Dawley rats a single, behavioural sensitizing dose of cocaine is sufficient to induce changes in the mRNA levels of growth-associated protein 43 (GAP-43), an important protein in mediating experience-dependent plasticity and synaptic reorganization, and of glutamate receptor 1 (GluR1), a subunit of AMPA glutamate receptors, a protein that is up regulated with repeated cocaine. Single exposure to 20, but not 10 mg/kg cocaine induced locomotor sensitization to a second injection of 10 mg/kg cocaine, observed at 24 h, 48 h and 7 days. Single dose of 20 but not 10 mg/kg cocaine 48 h before scheduled death significantly enhanced GluR1 and GAP-43 mRNA expression in the nucleus accumbens (NAc), both shell and core subregions, and ventral tegmental area (VTA). No changes were found in the levels of mRNA for GluR1 and GAP-43 in the frontal cortex, caudate putamen, dentate gyrus of hippocampus and basolateral nucleus of the amygdala after the single dose of 20 mg/kg cocaine. These results further strengthen the involvement of NAc and VTA in the behavioural sensitization and suggest a role of GAP-43 in the synaptic reorganization associated to drug abuse. PMID- 15548229 TI - Fast onset of dopamine uptake inhibition by intravenous cocaine. AB - In vivo voltammetry in the nucleus accumbens of anesthetized rats was used to investigate the time of onset of dopamine uptake inhibition by intravenous cocaine. There is disagreement between behavioral and neurochemical studies concerning the time-course of cocaine effects. Because of the high temporal resolution of voltammetry, the processes of dopamine release and uptake could be temporally separated to make evaluation of cocaine effects on uptake easier to address. Within 4 s after intravenous cocaine administration (1.5 mg/kg) there was significant inhibition of dopamine uptake that reached a plateau in 20 s. The peak heights of electrically evoked dopamine signals were also rapidly increased by cocaine. The signals returned to baseline values within approximately 1 h. In parallel behavioral studies, locomotor activity was significantly increased within 5-6 s following intravenous infusion of cocaine. Here we demonstrate that intravenous cocaine administration begins inhibiting the uptake of dopamine within a few seconds. This is at least 10-fold faster than previous neurochemical estimates. The present findings may contribute to the understanding of the neurobiological mechanisms underlying the early behavioral responses to cocaine. PMID- 15548230 TI - Dopamine, depression and antidepressants. AB - Abstract The relationship between depression and dopamine deficiency in the mesolimbic pathway has been hypothesized for many years. The experimental studies with animal models of depression and the human studies implicate the role of the dopamine system in depression. Not only do dopaminergic receptor agonists, but also antagonists such as olanzapine exhibit antidepressant effects associated with standard antidepressants in patients with treatment-resistant depression. This paradoxical result suggests that further investigations are necessary to understand the role played by dopamine in depression. PMID- 15548231 TI - Impact of inter-individual differences in drug metabolism and pharmacokinetics on safety evaluation. AB - Safety evaluation aims to assess the dose-response relationship to determine a dose/level of exposure for food contaminants below which no deleterious effect is measurable that is 'without appreciable health risk' when consumed daily over a lifetime. These safe levels, such as the acceptable daily intake (ADI) have been derived from animal studies using surrogates for the threshold such as the no observed-adverse-effect-level (NOAEL). The extrapolation from the NOAEL to the human safe intake uses a 100-fold uncertainty factor, defined as the product of two 10-fold factors allowing for human variability and interspecies differences. The 10-fold factor for human variability has been further subdivided into two factors of 10(0.5) (3.16) to cover toxicokinetics and toxicodynamics and this subdivsion allows for the replacement of an uncertainty factor with a chemical specific adjustment factor (CSAF) when compound-specific data are available. Recently, an analysis of human variability in pharmacokinetics for phase I metabolism (CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4, hydrolysis, alcohol dehydrogenase), phase II metabolism (N-acetyltransferase, glucuronidation, glycine conjugation, sulphation) and renal excretion was used to derive pathway-related uncertainty factors in subgroups of the human population (healthy adults, effects of ethnicity and age). Overall, the pathway-related uncertainty factors (99th centile) were above the toxicokinetic uncertainty factor for healthy adults exposed to xenobiotics handled by polymorphic metabolic pathways (and assuming the parent compound was the proximate toxicant) such as CYP2D6 poor metabolizers (26), CYP2C19 poor metabolizers (52) and NAT-2 slow acetylators (5.2). Neonates were the most susceptible subgroup of the population for pathways with available data [CYP1A2 and glucuronidation (12), CYP3A4 (14), glycine conjugation (28)]. Data for polymorphic pathways were not available in neonates but uncertainty factors of up to 45 and 9 would allow for the variability observed in children for CYP2D6 and CYP2C19 metabolism, respectively. This review presents an overview on the history of uncertainty factors, the main conclusions drawn from the analysis of inter-individual differences in metabolism and pharmacokinetics, the development of pathway-related uncertainty factors and their use in chemical risk assessment. PMID- 15548232 TI - Functional interaction of intestinal CYP3A4 and P-glycoprotein. AB - Intestinal CYP3A4-mediated biotransformation and active efflux of absorbed drug by P-glycoprotein are major determinants of bioavailability of orally administered drugs. The hypothesis that CYP3A4 and P-glycoprotein may act in concert to limit oral drug bioavailability is attractive from a theoretical point of view. Evidence in support of such an interplay between CYP3A4 and P glycoprotein comes mainly from a limited number of in vitro and animal studies. Obviously, it is a challenging task to demonstrate in vivo in humans that the function of CYP3A4 and P-glycoprotein in enterocytes is complementary, and results to directly support this concept remain elusive. However, CYP3A4 and P glycoprotein are clearly an integral part of an intestinal defence system to protect the body against harmful xenobiotics, and drugs that are substrates of both proteins often have a low bioavailability after oral administration. The functional interaction of intestinal CYP3A4 and P-glycoprotein warrants additional study. Further understanding this interplay would be potentially useful during drug development to solve bioavailability problems of new drug entities. PMID- 15548233 TI - Carvedilol attenuates ischemia-reperfusion-induced oxidative renal injury in rats. AB - There is increasing evidence to suggest that toxic oxygen radicals play a role in the pathogenesis of ischemia/reperfusion (I/R) injury in the kidney. This study was designed to investigate the effects of carvedilol (CVD), an antihypertensive drug in I/R-induced renal failure in rats. The protective effect of CVD against the damage inflicted by reactive oxygen species (ROS) during renal I/R was investigated in Sprague-Dawley rats using histopathological and biochemical parameters. In one set of experiments, animals were unilaterally nephrectomized, and subjected to 45 min of left renal pedicle occlusion and in another set both the renal pedicles were occluded for 45 min followed by 24 h of reperfusion. Carvedilol (2 mg/kg, i.p.) was administered twice, 30 min prior to ischemia and 12 h after the reperfusion period. At the end of the reperfusion period, rats were killed. Thiobarbituric acid-reactive substances (TBARS), reduced glutathione (GSH) levels, catalase (CAT) and superoxide dismutase (SOD) activities were determined in renal tissue. Serum creatinine and blood urea nitrogen (BUN) concentrations were measured for the evaluation of renal function. Ischemic control animals demonstrated severe deterioration of renal function, renal morphology and a significant renal oxidative stress. Pretreatment of animals with CVD markedly attenuated renal dysfunction, morphological alterations, reduced elevated TBARS levels and restored the depleted renal antioxidant enzymes. The findings imply that ROS play a causal role in I/R-induced renal injury and CVD exerts renoprotective effects probably by the radical scavenging and antioxidant activities. PMID- 15548234 TI - Idazoxan effects upon contractile activity in the rat aorta are related to alpha adrenoceptors and L-type channels. AB - Idazoxan is an alpha(2) adrenoceptor antagonist and alpha(1)/alpha(2) partial agonist which also blocks imidazoline receptors. Although idazoxan is widely used in pharmacological studies, its intrinsic vasoactive properties could bring about some limitations. Others have shown that in rat aorta contracted by phenylephrine idazoxan induces relaxation and that in rat small arteries it preferentially antagonizes the alpha(1)-mediated response. We further investigated this matter, using the rat aorta and focusing on the endothelium-independent effects and on L type channels. In our study, idazoxan inhibited the contraction induced by phenylephrine, an effect which was stronger in the presence of endothelium, but did not affect the contractions induced by various other agents (high potassium, angiotensin II, prostaglandin F(2alpha)). This preferential inhibition was attenuated by 10(-4) m, but not by 10(-5) m yohimbine, and also reduced by 10(-2) m tetraethylammonium and blunted by 10(-4) m methoxyverapamil. In concentrations above 10(-5) m idazoxan induced weak contractions of the de-endothelized rings, which were prazosin- and methoxyverapamil-sensitive. Others have suggested that cyclic guanosine monophospate mediates the idazoxan-induced endothelium-dependent relaxation, but this is difficult to reconcile with our findings. Potassium efflux could play some role in the direct relaxing effect of idazoxan. The observed idazoxan effects appear as based on action upon alpha(1) receptors, but a direct interaction with L-type calcium channels could also be taken into consideration. PMID- 15548235 TI - Relationship between noradrenaline and nonlinear indexes of blood pressure dynamics in normotensive and spontaneously hypertensive rats. AB - Recent studies in normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats show that some nonlinear indexes derived from the recurrence plot method may be better markers of sympathetic activity than the spectral powers of blood pressure (BP). We herein investigated the relationships between nonlinear indexes and plasma noradrenaline concentration in conscious WKY rats and SHRs. Blood pressure was recorded for 30 min after intravenous injection of saline (0.9% NaCl, 100 microL/kg), hexamethonium (20 mg/kg), atropine (0.5 mg/kg), atenolol (1 mg/kg) or prazosin (1 mg/kg). Spectral power in the low-frequency (LF) band and the nonlinear index (L(max)), calculated on diastolic (DBP) and systolic blood pressures (SBP), were used to analyse the BP signal. Noradrenaline concentration was determined by radioenzymatic technique. A robust stepwise regression analysis - using noradrenaline concentration as dependent variable, and LF, L(max) and treatment, as independent variables -shows that treatment is the main variable explaining the variance of noradrenaline level in WKY rats, excluding the use of the pooled data to explore the relationship between noradrenaline concentration and LF or L(max). In contrast, in SHRs, treatment has no effect on the variance of noradrenaline concentration and the pooled data were then used. In this group, no correlation was observed between noradrenaline concentration and LF. In contrast, very high positive correlation was observed between noradrenaline level and L(max)-DBP (r = 0.59; P = 0.0005) or L(max)-SBP (r = 0.53; P < 0.002). The results strengthen our previous suggestion that nonlinear indexes may be better tools than spectral powers to investigate the sympathetic nervous system. PMID- 15548236 TI - Cardiovascular and survival effects of sympatho-inhibitors in adriamycin-induced cardiomyopathy in rats. AB - Adriamycin (ADR) is a widely used drug for the treatments of cancers. This study evaluates the effects of moxonidine and metoprolol on cardiac hemodynamics and survival in ADR-induced left ventricular dysfunction (total dose of 20 mg/kg in a 4-week regimen). Rats were treated with the centrally acting I(1)R agonist sympatho-inhibitor, moxonidine, or with the non-selective beta-adrenergic antagonist, metoprolol, during 1 month or until death. Treatments began 1 week after the onset of the ADR administration. Low doses (0.5 and 1 mg/kg/day) of moxonidine and metoprolol (10 mg/kg/day) improved cardiovascular function. High doses of moxonidine (3 mg/kg/day) and metoprolol (150 mg/kg/day) were cardiodepressive. Moxonidine and metoprolol both failed to improve survival. These data indicate that a treatment with these sympatho-inhibitors can reduce the left ventricular dysfunction induced by ADR. Moreover, these cardioprotective effects where obtained even when ADR was used at a dose regimen usually employed for its antineoplastic effects in rodents. Nevertheless, in this particular cardiomyopathy, we did not find any association between improvements of functional parameters and survival whatever the drug and the dose used. This problem points out the difficulty to prevent, at least with sympatho-inhibitory drugs alone, the mortality linked to the chronic cardiotoxicity of ADR. PMID- 15548237 TI - Myocardial infarction non-invasively induced in rabbits by administering isoproterenol and vasopressin: protective effects exerted by verapamil. AB - Myocardial infarction is usually induced in small animals by means of invasive procedures: the aim of this study was to cause heart necrosis lesions by non invasive means. We injected rabbits with isoproterenol (3 mg/kg, i.p.) and vasopressin (0.3 mg/kg/5 min, i.v.) alone and in combination, and studied their effects on myocardial histology, electrocardiographic profiles, the appearance of the plasma cardiac necrosis marker c-troponin I (c-TPN I), hemodynamic parameters (blood pressure, heart rate), the coagulative process partial throboplastine time (PTT), and plasma nitric oxide (NO) levels. In the rabbits treated with vasopressin alone, the ischemic damage was associated with a decrease in NO values, and the appearance of electrocardiographic T-wave inversion and low plasma c-TPN I levels, whereas the animals treated with isoproterenol alone had necrotic bands in the myocardium, plasma c-TPN I, and electrocardiographic modifications (ST-segment changes and T-wave inversion). Combined treatment increased myocardial alterations such as contraction band necrosis, induced the appearance of specific hypoxic lesions such as areas of coagulative necrosis and leukocyte infiltration, and led to higher plasma c-TPN I levels and altered ECG profiles. Both drugs favored a decrease in plasma NO values and further alterations in hemodynamic parameters, such as higher blood pressure and greater procoagulant activity. The myocardial necrosis and modified cardiovascular parameters were attributed to calcium activated processes and the decrease in NO levels. As this model of myocardial damage involves the use of drugs that facilitate the opening of L-calcium channels, we also investigated their effects on cardiovascular parameters and heart histology after pretreatment with the calcium antagonist verapamil; this drug protected against the appearance of histological myocardial lesions, electrocardiographic alterations and high plasma c-TPN I levels, and prevented the hemodynamic and procoagulation changes, but did not affect the decrease in plasma NO values. The protective effects were attributed to the drug's calcium antagonist activity. In conclusion, the injection of isoproterenol and vasopressin induces a myocardial infarction non invasively and seems to be suitable for studying early myocardial ischemic lesions and the effects of drugs interfering with myocardial damage and its related phenomena. PMID- 15548238 TI - Impairment of smooth muscle function of rat thoracic aorta in an endothelium independent manner by long-term administration of N(G)-nitro-L-arginine methyl ester. AB - In this study, we aimed to elucidate whether the daily hypertensive dose of long term N(G)-nitro-l-arginine methyl ester (l-NAME) treatment, could make a difference between endothelial and smooth muscle functions in rat thoracic aorta. We test the hypothesis that high-dose, long-term l-NAME treatment has a depressive effect on vascular smooth muscle contractile activity which is not related with nitric oxide (NO) synthesis inhibition. After 14 days of treatment, isometric tension and (45)Ca(2+) influx were measured in aortic tissues isolated from l-NAME(10) and l-NAME(100) hypertensive (10 and 100 mg/kg/day, systolic blood pressures 167 +/- 7 and 172 +/- 10 mmHg, respectively) and control normotensive rats (132 +/- 7 mmHg). In l-NAME(10)- and l-NAME(100)-treated rats, acetylcholine-induced relaxation in aortic rings was suppressed with no significant difference between the treatments. l-NAME(100) (but not l-NAME(10)) treatment, significantly inhibited contractile responses to phenylephrine, angiotensin II, and K(+) (80 mm) in endothelium-intact tissues. The effect of l NAME(100) on phenylephrine-induced contractile responses was not observed after 3 days of treatment. In endothelium-denuded aortic tissues of l-NAME(100) (but not l-NAME(10))-treated rats, phenylephrine (1 x 10(-6) m)- and K(+) (80 mm)-induced contractions and (45)Ca(2+) influxes were significantly reduced. In Ca(2+)-free medium (0.1 mm EDTA), on the contrary, the transient contractions obtained by either phenylephrine (1 x 10(-6) m) or caffeine (1 x 10(-2) m), or the sustained contractions induced by 12-o-tetradecanoylphorbol-13-acetate (1 x 10(-6) m; a protein kinase C activator) in endothelium-denuded aortic rings, were not modified by both l-NAME treatments. These results indicate that in aortic rings from l-NAME hypertensive rats, low and high doses, long-term l-NAME administration may be associated with equivalent inhibition in NO-dependent vasodilator tone (corresponding to equivalent hypertension values); whereas only high-dose, long-term l-NAME administration produces an endothelium-independent decrease in vasocontrictor activity, at least partly explained by a reduction in extracellular Ca(2+) influx. PMID- 15548239 TI - Decreased aortic smooth muscle contraction in a rat model of multibacterial sepsis. AB - We investigated whether blockade of the smooth muscle cell (SMC) inducible nitric oxide synthase (iNOS)-soluble guanylyl cyclase (sGC) vasodilator pathway would restore the fall in vasoreactivity produced by sepsis following cecal ligation and perforation (CLP) in rats. Contraction of adjacent aortic rings paired for the presence or absence of endothelial cells (EC) was recorded following high [K(+)](e) (40 mm) or norepinephrine (NE, 10(-8) to 10(-5) m) in the presence of the nitric oxide synthase inhibitor (NOS), N(G)-nitro-l-arginine methyl ester (l NAME, 0.3 mm) or the sGC inhibitor, 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1 one (ODQ, 5 mum). In EC-denuded rings, sepsis halved SMC contraction induced by high [K(+)](e) or NE; neither l-NAME nor ODQ produced an increase in NE E(max) or high [K(+)](e)-evoked contraction. In conclusion, SMC contractility is globally reduced in CLP; this reduction does not appear to be explained by induction of SMC NOS in this CLP model. PMID- 15548240 TI - Does rat global transient cerebral ischemia serve as an appropriate model to study emotional disturbances? AB - We used two validated psychopharmacological methods, the forced swimming test (FST 20 min and 5 min) and the elevated plus-maze (EPM), to quantify depression like and anxiety-like behavior induced by transient global cerebral ischemia in the rat. We also validated use of these methods for the study of antidepressant (imipramine) and anti-anxiety drugs (diazepam). Twelve days after surgery to provoke transient global ischemia, spontaneous motor activity was 40% higher in ischemic rats than in sham-operated controls. Duration of immobility during the FST 20 min and 5 min was 28 and 30% shorter, respectively, than in controls. Treatment with imipramine (3 x 30 mg/kg i.p.) induced a significantly shorter duration of immobility during the FST 5 min, but with no difference between ischemia and control rats. The EPM demonstrated that ischemia did not induce any change in the six behavior parameters measured. Diazepam (1.5 mg/kg i.p.) induced significant anxiolytic effects which were similar in ischemic and sham-operated animals. Both tests failed to demonstrate perturbed performance but conversely, these findings did disclose the sensitivity of ischemia-exposed rats to the action of imipramine and diazepam, demonstrating the usefulness of these tests as psychopharmocological tools for evaluating the effect of psychotropics in the ischemic rat. PMID- 15548241 TI - Effects of venlafaxine on ethanol withdrawal syndrome in rats. AB - The present study was designed to investigate the effects of venlafaxine, a serotonin and noradrenaline reuptake inhibitor (SNRI), on ethanol withdrawal syndrome in rats. Adult male Wistar rats (187-319 g) were used for the study. Ethanol (7.2%, v/v) was given to rats by a liquid diet for 21 days. Control rats were pair-fed an isocaloric liquid diet containing sucrose as a caloric substitute to ethanol. Venlafaxine (5, 10, 20 and 40 mg/kg) and saline were injected to rats intraperitoneally just before ethanol withdrawal. After the 2nd, 4th and 6th hour of ethanol withdrawal, rats were observed for 5 min, and withdrawal signs that included locomotor hyperactivity, agitation, stereotyped behaviour and wet dog shakes were recorded or rated. A second series of injections was given at the 6th hour after the first one, and rats were then tested for audiogenic seizures. Venlafaxine produced some inhibitory effects on locomotor hyperactivity, stereotypic behaviours and wet dog shakes. However, a two-way anova of the data did not indicate any significant effect. It reduced the incidence of the audiogenic seizures at the 6th hour of ethanol withdrawal. Venlafaxine (20 mg/kg) also prolonged the latency of the seizures significantly. Our results suggest that acute venlafaxine treatment has limited beneficial effects on ethanol withdrawal syndrome in rats. PMID- 15548242 TI - A QSAR analysis of toxicity of Aconitum alkaloids. AB - Biological activity of Aconitum alkaloids may be related to their toxicity rather than to a specific pharmacological action. A Quantitative structure-activity relationships (QSAR) analysis was performed on the following two groups of alkaloids: compounds with an aroyl/aroyloxy group at R(14) position (yunaconitine, bulleyaconitine, aconitine, beiwutine, nagarine, 3-acetyl aconitine, and penduline), and compounds with the aroyloxy group at R(4) position (N-deacetyllappaconitine, lappaconitine, ranaconitine, N-deacetylfinaconitine, N deacetylranaconitine). The LD(50) (micromol/kg) of the 12 alkaloids were obtained from the literature. LD(50) was significantly lower in group 1 than in group 2. The steric and core-core repulsion energies were significantly higher in group 1. The total energy and heat of formation and electronic energies were significantly lower in group 1. The reactivity index of N, C1', C4' and C6' were similar between groups. The reactivity index of C2' was significantly higher and the reactivity index of C3' and C5' were significantly lower in group 1. Log P and pKa were similar between groups. Molecular weight was significantly higher in group 1. A significant linear relationship was observed between log LD(50) and either analgesic log ED(50) or local anesthetic log ED(50). The LD(50)/analgesic ED(50) obtained from average values was 5.9 for group 1 and 5.0 for group 2. However, the LD(50)/local anesthetic ED(50) was 40.4 and 318, respectively. The study supports that the analgesic effects of these alkaloids are secondary to their toxic effects whereas alkaloids from group 2 are susceptible to be further studied as local anesthetic agents. PMID- 15548243 TI - Pharmacogenomics in cardiovascular clinical trials. AB - Genomics - having quickly emerged as the central discipline in basic science and biomedical research - is poised to take the center stage in clinical medicine as well over the next few decades. Although there is no specific regulatory guideline on the application of pharmacogenetics to drug development, some recommendations are already included in several published guidelines on drug development. The patients more likely to provide the most valuable information on the specific contribution of a given gene or its variant are those who fail to respond to a drug ('therapeutic failures') and those who develop toxicity to the drug. However, before drawing definite conclusions on subgroups following pharmacogenomic analyses, one must be aware of disease classification, data collection, and how much is known about the disease process. It seems reasonable to collect genomic DNA from all patients enrolled in clinical drug trials (along with appropriate consent to permit pharmacogenetic studies) for the purpose of post hoc analyses. One exception to post hoc genomic analysis is when patients with a specific genotype are excluded from randomization into a clinical trial. Physicians will need to understand the concept of genetic variability, its interactions with the environment (e.g. drug-drug or drug-disease interactions), and its implication for patient care. PMID- 15548244 TI - Effect of preoperative oral sustained-release morphine sulfate on postoperative morphine requirements in elective spine surgery. AB - Sustained-release morphine sulfate (SRMS) is a painkiller used in oncology. The purpose of our study was to assess its efficacy on postoperative morphine requirements in elective spine surgery. This was a placebo-controlled, randomized, double-blind study. Adults scheduled for spine surgery under general anesthesia were orally administered SRMS (30 mg) or a placebo 2 h before surgery. Primary endpoint was postoperative cumulated morphine consumption through patient controlled analgesia (PCA) during the 12 h following extubation. Statistical analysis was performed using a sequential method, the triangular test. The study was stopped after the sixth analysis (51 patients had been included; placebo: 26, SRMS: 25). Age, weight, sex ratio, type of surgery, intra-operative sufentanil consumption, anesthesia duration and time to extubation were similar in the two groups. Morphine consumption through PCA during the 12 h following extubation was significantly lower in the SRMS group (mean +/- SD: 10.5 +/- 7.6 mg) compared with placebo group (15.6 +/- 6.0 mg, P = 0.016, sequential analysis). Corresponding unbiased median estimates were 10.6 and 15.8 mg in SRMS and placebo groups, respectively. Morphine consumption through PCA during the 24 h following extubation was also significantly lower in the SRMS group (15.9 +/- 12.7 mg) compared with the placebo group (23.8 +/- 10.9 mg, P = 0.032). Vigilance, nausea and respiratory rate 3 and 6 h after extubation were similar in the two groups. A preoperative oral administration of SRMS (30 mg) induces a 33% reduction of postoperative morphine requirements in patients scheduled for spine surgery without inducing side effects. PMID- 15548245 TI - Patient information after ruptured intracranial aneurysm. AB - AIM: This paper reports an investigation into the effects of increased information for patients treated for intracranial aneurysm rupture. BACKGROUND: Intracranial aneurysm rupture is a grave condition that requires immediate care. It can be treated in two different ways, by surgery or by endovascular procedure. Intracranial aneurysm rupture can mean great changes in life, both for the patient and their spouse or relatives. METHODS: An intervention study was conducted using a quasi-experimental design. Participants were recruited consecutively over a period of 12 months and consisted of 62 patients treated for intracranial aneurysm rupture at a Swedish neurosurgical clinic. They were divided into two groups: an intervention group, who received written and oral information, and a comparison group, who received only oral information. INSTRUMENTS: Self-report questionnaires were sent to patients' homes 1-3 months after the aneurysm rupture. The questionnaires consisted of one study-specific instrument with questions about understanding of the information given and the State-Trait Anxiety Inventory, which measures worry or anxiety. FINDINGS: The intervention group considered that the information that they received was somewhat easier to understand and that it corresponded more closely to their needs, compared with the comparison group. The majority of patients in both groups expressed a need for more and improved information. Levels of anxiety were high for the majority of patients, but no significant difference was evident between groups. Furthermore, the results showed that the majority of patients were given information without their spouse or relatives being present. CONCLUSION: Increased information seems to be needed for these patients. There is a need to continue the work to improve information-giving to them and their relatives. PMID- 15548246 TI - Postoperative mobilization of patients with abdominal aortic aneurysm. AB - AIM: This paper reports on a study which aimed to evaluate the effects of structured written preoperative information on patients' postoperative psychological and physical wellbeing after surgery for abdominal aortic aneurysm (AAA). BACKGROUND: The possible benefits of current booklets written by professionals on postoperative psychological and physical wellbeing in patients with AAA are unknown. Previous studies have shown that preoperative information has a favourable effect on both mood state and physical mobilization. METHOD: Fifty-two patients admitted for elective repair of AAA were selected consecutively and randomized to receive only verbal (control group), or verbal and written information in booklet form (experimental group). The booklet contained procedural and sensory information about the disease and its treatment. Two questionnaires were used to establish whether the booklet had any effect on perceived health, psychological and physical wellbeing postoperatively. RESULTS: The two groups were similar regarding their perceived health but differed significantly regarding psychological wellbeing pre- and postoperatively. Patients in the experimental group were significantly sadder both pre- and postoperatively compared with those in the control group. Both groups were similar in postoperative physical wellbeing. CONCLUSION: This group of patients often has asymptomatic disease, with a short interval between diagnosis and major surgery. When patients receive an information booklet during this period, this seems to cause more worries than anticipated. Hence, a more supportive educational programme might benefit this patient group, both pre- and postoperatively. PMID- 15548247 TI - Effects of absorbent incontinence pads on pressure management mattresses. AB - AIM: The aim of this paper is to report a study to determine the effects of absorbent pads on the pressure-relieving properties of 'standard' and pressure management mattresses. BACKGROUND: Pressure ulcers and incontinence often co exist. There is a strong association between poor mobility and continence problems and patients using pressure management products are therefore also likely to be using absorbent pads. METHODS: An instrumented articulated anthropometric phantom with simulated soft body 'tissues' in the gluteal and sacral areas was used as the 'subject'. The phantom was raised and lowered on to three mattresses (standard foam, visco-elastic foam and surface-cut foam) in three states: naked, wearing a dry pad (Tena Super, SCA Hygiene AB) and wearing a wet pad. The pressure mapping device Xsensor was used to record the distribution of pressure over the sacral and ischial areas of the phantom. Peak pressure was used as the primary outcome variable and 10 repeats were made on each mattress under each condition. RESULTS: There were substantial and significant differences for all three mattresses in recorded peak pressures between the naked buttocks and the buttocks wearing a dry pad. There were no significant differences between measurements made using the dry vs. wet pad. Peak pressures frequently occurred over areas of pad folds. Additional testing showed that pads that were 'smoothed' by hand had significantly lower peak pressures than 'unsmoothed' pads. CONCLUSIONS: Absorbent pads have a substantial adverse effect on the pressure redistribution properties of mattresses. Pad folds appear to contribute to this effect, which can be ameliorated slightly by smoothing. Absorbent pad manufacturers should consider engineering pads that minimize disruption to pressure management. Further examination of continence and pressure management products is necessary to establish optimum combinations for nursing care. PMID- 15548248 TI - Promoting health for families of children with chronic conditions. AB - AIM: This paper reports a study whose aim was to describe how nurses in a paediatric unit promote the health of families of children with chronic conditions during the children's hospitalization. BACKGROUND: It is stressful for a family when a child has a chronic condition and hospitalized. Nurses have the opportunity to promote family health while a child is hospitalized, and previous studies contain a wealth of recommendations on how this should be done. However, there is little research evidence about family care and how nurses promote the health of a family with a chronically ill child. METHODS: Forty nurses who worked in two paediatric units providing care for children with chronic conditions and their families participated in the study. Data were collected in two Finnish hospitals in 2002 using group interviews, and nursing actions were observed in both units for a period of 84 hours. Data analysis was based on the grounded theory method, proceeding to the stage of axial coding. Data were collected until theoretical saturation was reached. FINDINGS: Family care was based on three distinctive strategies. Nurses used a systematic, selective or situation-specific strategy while promoting family health during a child's hospitalization. CONCLUSIONS: The findings indicate that nurses used different strategies while promoting family health during a child's hospitalization. The systematic way of working with families identified in the study seems a useful strategy. All three strategies of family nursing described here should be tested in other paediatric wards and in other geographical locations. PMID- 15548249 TI - Quality of life and symptoms among older people living at home. AB - AIM: This paper reports a study comparing the socio-demographic data, quality of life (QoL) and symptoms of older people living at home with and without help. BACKGROUND: Despite growing numbers of older people worldwide, little is know about the differences between older people receiving help to live at home and those not receiving this, especially as regards QoL and symptoms. Not only symptoms but also dependency on others per se may reduce older people's QoL. From a nursing perspective, knowledge about such issues is important because the impact of symptoms may be reducible, even when diseases cannot be cured. METHOD: A postal questionnaire was sent to an age-stratified random sample of 1866 people aged 75 years or over. Of the respondents (n = 1248) 448 received help and 793 did not. RESULTS: The group receiving help had a significantly higher age, more women, more people widowed and living alone, more children, a higher number of self-reported diseases and symptoms, greater inability to remain alone at home and lower QoL. Loneliness, depressed mood and abdominal pain were significantly related to low QoL in both groups. Living alone, not being able to remain alone at home without help, and fatigue were also predictive of low QoL among those receiving help, and number of diseases and sleep problems in those without. CONCLUSION: Receiving help with daily living seems to be significantly related to low QoL and goes along with a high number of symptoms that need to be considered in nursing care. Through regular visits, systematic assessment and intervention, especially focusing on older people's symptoms, nurses may contribute to improved QoL for this section of the population. PMID- 15548250 TI - Multicomponent individualized smoking cessation intervention for patients with lung disease. AB - AIMS: This paper reports a study examining the process and outcomes of a long term, multicomponent smoking cessation intervention for patients with lung disease initiated while hospitalized and provided over 1-year postdischarge. BACKGROUND: Successful smoking cessation interventions are of primary importance for people with lung disease. Initiation of such an intervention in hospital settings is particularly important as patients may be especially motivated to quit as a result of strong perceptions of vulnerability while hospitalized for a smoking-related disease. Tailoring the intervention to each person's needs is a promising approach to practice. METHODS: All patients who smoked and were admitted to a pulmonary unit over 2 years were invited to participate in this quasi-experimental study (n = 85), and 69 continued beyond the first month. The intervention was shaped by the TransTheoretical Model and used nicotine replacement therapy, along with individual and group counselling and support grounded in the nurse-patient relationship. The intervention was provided during hospitalization and by telephone after discharge at 1 week, and 1, 3, 6 and 12 months. RESULTS: At 12-months postdischarge, 39% of the patients reported continuous abstinence from smoking from the time they joined the programme and 52% were not smoking at that time. No relationship was found between abstinence and the number of quit attempts, readiness to quit, nicotine dependency and length of hospital stay. Readiness to quit had increased and nicotine dependency decreased significantly by the end of the programme. No gender differences were found for the main variables. CONCLUSIONS: Comprehensive, individualized smoking cessation interventions for hospitalized patients having lung disease, with a 1 year follow-up, was successful. Abstinence was high in comparison with other studies. This may in part be explained by significantly enhanced motivation to quit during the smoking cessation programme. PMID- 15548251 TI - Grounded theory: reflections on the emergence vs. forcing debate. AB - AIM: The aim of this paper is to compare Glaser's model of theory generation, where theory rises directly and rigorously out of the data, devoid of interpretivism, to Strauss's conceptually descriptive approach that encourages directive questioning and supports an interpretive stance. BACKGROUND: The discovery of grounded theory (GT) was born out of a merger between Barney Glaser and Anselm Strauss, the proverbial 'fathers' of GT. Since the co-creation of their approach to theory development through research in 1967, these scholars have taken seemingly divergent paths in further developing and evolving the pragmatic use of GT. DISCUSSION: Numerous researchers have used GT as a general method, applying it to both quantitative and qualitative research approaches. In this paper we discuss the stages and strategies of data sampling, collection, coding and analysing used by both Glaser and Strauss. Constant comparative analysis is identified as the primary strategy in the integrated coding and analysing stages of this theorizing method, regardless of the researcher's philosophical or research orientation. We also discuss initial or open coding, advanced coding, memoing, and theoretical sampling, with particular attention to comparing and contrasting the descriptive terms and application strategies that have been suggested by both Glaser and Strauss. CONCLUSION: The reported distinctions in the approach, method, and general intent of GT reflected in this paper are not easy to comprehend. The two methods reflect different basic philosophical paradigms, and therefore represent distinct approaches to GT. Researchers need to be clear about which philosophy and resulting analysis approach they are using, and the effect that approach will have on the research process and outcomes. PMID- 15548252 TI - Validation of pressure ulcer risk assessment scales: a critique. AB - AIM: The aim of this paper is to present critical analysis of the validation methods of pressure ulcer risk assessment scales. BACKGROUND: The validation of pressure ulcer risk assessment scales remains a topic of considerable debate and uncertainty. The Braden scale and Norton scale are the most frequently used. Sensitivity and specificity are the recommended and most commonly used epidemiological tools to evaluate the validity of those risk assessment scales. DISCUSSION: The use of preventive measures influences both the sensitivity and specificity of the scales. Analysis of published studies on risk assessment scales reveals that, although some patients received preventive measures and others did not, this was not taken into account. Consequently, generalization of those results is not possible. Some possible alternative designs for studying the validity of risk assessment scales are discussed. CONCLUSIONS: Currently available risk assessment scales are of only limited value, and there use will result in many patients being falsely identified as at risk or not at risk. Sensitivity and specificity criteria are not the most appropriate tools to validate risk assessment scales. A risk assessment scale should be evaluated in combination with the preventive measures used. PMID- 15548253 TI - Stressors, burnout and social support: nurses in acute mental health settings. AB - AIMS: This paper reports a study which aims (1) to investigate and compare levels of stressors and burnout of qualified and unqualified nursing staff in acute mental health settings; (2) to examine the relationships between stressors and burnout and (3) to assess the impact of social support on burnout and stressor burnout relationships. BACKGROUND: Several studies have noted that the work of mental health nurses can be highly stressful, but relatively few have focused specifically on staff working in acute inpatient settings. Although many of the pressures faced by this group are similar to those in other nursing specialties, a number of demands relate specifically to mental health settings, including the often intense nature of nurse-patient interaction and dealing with difficult and challenging patient behaviours on a regular basis. METHODS: A convenience sample of 93 nursing staff from 11 acute adult mental health wards completed the Mental Health Professionals Stress Scale, Maslach Burnout Inventory and House and Wells Social Support Scale. RESULTS: Lack of adequate staffing was the main stressor reported by qualified staff, while dealing with physically threatening, difficult or demanding patients was the most stressful aspect for unqualified staff. Qualified nurses reported significantly higher workload stress than unqualified staff. Approximately half of all nursing staff showed signs of high burnout in terms of emotional exhaustion. A variety of stressors were positively correlated with emotional exhaustion and depersonalization. Higher levels of support from co workers were related to lower levels of emotional exhaustion. Higher stressor scores were associated with higher levels of depersonalization for staff reporting high levels of social support, but not for those reporting low levels of support (a reverse buffering effect). CONCLUSIONS: Qualified and unqualified nursing staff differed in terms of the prominence given to individual stressors in their work environment. The findings were consistent with the notion of burnout developing in response to job-related stressors. While staff support groups may be useful in alleviating feelings of burnout, the reverse buffering effect suggests that they should be structured in a way that minimizes negative communication and encourages staff to discuss their concerns in a constructive way. PMID- 15548254 TI - Predictors of life satisfaction of Korean nurses. AB - AIM: This paper reports a study to identify the levels of work satisfaction, burnout and life satisfaction among Korean hospital nurses and the relative importance of negative and positive work outcomes (burnout and work satisfaction) in explaining the variance of life satisfaction of nurses. BACKGROUND: Previous research has demonstrated that work outcomes such as job satisfaction and burnout can affect overall life satisfaction. It is not yet known, however, whether positive and negative aspects of work outcomes exert equally strong or varying degrees of effect, nor whether the relationships demonstrated primarily in Western cultures also are present in other cultures. Given the widespread shortage of nurses, it is important to identify areas for organizational interventions that have the greatest potential for improving both recruitment and retention of nurses. METHODS: A cross-sectional correlational design was used. A survey was undertaken with 194 nurses from general hospitals of 300 beds or more in southern Korea between May 1999 and March 2000. Paper and pencil self-rating questionnaires were used to gather information. Data were analysed using descriptive statistics, Pearson correlations and multiple regressions. RESULTS: Korean nurses reported moderate levels of life satisfaction, with low levels of work satisfaction and high levels of burnout. Burnout explained more variance in life satisfaction than work satisfaction. Those who experienced higher personal accomplishment and lower emotional exhaustion and who were satisfied with their professional status and did not work at night reported higher life satisfaction. CONCLUSIONS: This study highlights the relative importance of negative work outcomes on nurses' overall well-being. Strategies to reduce emotional exhaustion, enhance nurses' personal accomplishment and satisfaction with professional status, and accommodate shift preferences for work scheduling were suggested. Replicating this study with nurses from other geographic areas using random selection will be needed to increase the generalizability of the findings. PMID- 15548255 TI - Professional career development for male nurses. AB - AIM: The aim of this paper is to report a study to: (a) explore Taiwanese male nurses' motivations for becoming a nurse; (b) reveal their professional developmental process in nursing; (c) understand the difficulties hindering their professional development from both professional and gender aspects; and (d) identify the strategies they use to cope with these difficulties. BACKGROUND: Hindered by historical, cultural, economic and warfare factors, the proportion of male nurses in Taiwan remains low. Taiwanese male nurses' career development process has not been well investigated yet. METHODS: A descriptive qualitative research design was used, with a convenience sample of 15 male nurses (mean age 30.8 years) with a Bachelor's degree in Nursing Science. Data were collected by semi-structured interviews and analysed by content analysis. RESULTS: Taiwanese male nurses' entrance into the nursing profession involved three phases: pre study, study and employment. The difficulties encountered during career development were related to the gender expectations of patients and the general public. The nurses received support more from superiors than from colleagues. The strategies they used included (a) improving their professional knowledge and skills to obtain higher levels of satisfaction and better opportunities for promotion; (b) thinking aggressively about job promotion; (c) choosing specialist departments as appropriate environments for graduate study and personal growth; and (d) changing their professional track for personal growth. CONCLUSIONS: Based on the nature of nursing work and clinical experiences, Taiwanese male nurses believed that nursing was a profession suitable for both men and women. Their preparation for career development started at the pre-study phase. The major strategies they used were related to a strong desire for personal growth and professional promotion. Finally, a conceptual framework was developed to depict this complex phenomenon. PMID- 15548256 TI - Strong foundations built to increase access to AIDS treatment. PMID- 15548260 TI - A transmembrane protein-tyrosine phosphatase receptor type C (CD45) exon A point mutation (77 C to G) is not associated with the development of type 1 diabetes mellitus in a German population. AB - To investigate whether a C to G transversion at position 77 in exon A of the CD45 gene is associated with the development of diabetes mellitus type 1 (T1D), we studied 165 patients and 220 control individuals. The frequency of the 77G allele in the control group was 1.1%, which was not significantly different from the 1.2% found in the patient group (P = 0.922). The C to G transversion does not seem to be associated with susceptibility for T1D. PMID- 15548261 TI - The CCR5Delta32 allele is associated with reduced liver inflammation in hepatitis C virus infection. AB - CCR5Delta32 is a deletion mutation in the chemokine receptor CCR5. Liver inflammatory activity was found to be significantly reduced (P = 0.005) in Jewish Israeli patients infected with the hepatitis C virus (HCV) carrying the CCR5Delta32 allele. The CCR5Delta32 allele does not alter susceptibility to HCV infection; however, it may play a role in the progression and outcome of the disease. PMID- 15548262 TI - Establishment of T cell-specific and natural killer cell-specific unigene sets: towards high-throughput genomics of leukaemia. AB - We report the establishment of highly non-redundant unigene sets consisting of cDNA clones derived from T lymphocytes and natural killer cells. Each set consists of 10 506 and 13 409 clones, respectively, arrayed on nylon membranes in duplicate. The sets provide an excellent tool for genome-wide gene expression analysis studies in immunology research. PMID- 15548263 TI - Lack of a strong association between HLA class II, tumour necrosis factor and transporter associated with antigen processing gene polymorphisms and virological response to alpha-interferon treatment in patients with chronic hepatitis C. AB - The aim of the study was to investigate whether polymorphisms of the HLA class II, tumour necrosis factor (TNF) and transporter associated with antigen processing (TAP) genes influence the response to alpha-interferon in patients with chronic hepatitis C. Twenty-seven sustained responders and 55 non-responders to alpha-interferon monotherapy were investigated. HLA-DRB1, DQA1, DQB1, TNFA, TNFB, TAP1 and TAP2 alleles were determined by PCR-based molecular techniques. Sustained virological response was defined as undetectable serum hepatitis C virus (HCV) RNA for at least 3 years after the end of treatment. Probability (P) values were corrected for the number of alleles tested (Pc). Viral genotype 1b was more frequent in responders than in non-responders (56% vs. 26%, P = 0.009). HLA-DQB1*02 occurred less frequently in responders than in non-responders (14.8% vs. 29%, Pc not significant). HLA-DRB1*11 and DQB1*0602 were found in 22.2% and 9.3% of responders and in 10.9% and 1.8% of non-responders, respectively (Pc not significant). There was no difference in the distribution of TNF alleles in the two groups. Twenty-four (88.8%) responder patients as compared with 34 (61.8%) non-responders were TAP1*0101 homozygous (Pc not significant). Thus, in European Caucasoids with chronic hepatitis C, we could not demonstrate a strong association between HLA class II, TNF, and TAP gene polymorphisms and response to interferon treatment. PMID- 15548264 TI - Immunoregulatory cytokine polymorphisms in Italian patients affected by paroxysmal nocturnal haemoglobinuria and aplastic anaemia. AB - We investigated regulatory variants of five cytokine genes [tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, transforming growth factor (TGF) beta, interleukin (IL)-6 and IL-10] in 40 Italian patients affected by paroxysmal nocturnal haemoglobinuria (PNH) and aplastic anaemia (AA). Genotypes associated with high production of TGF-beta and IFN-gamma were more frequent in patients than in controls. Genetic regulation of the immunological pathways involved in the pathogenesis of bone marrow failure is suggested. PMID- 15548265 TI - Presence of HLA antibodies in single-donor-derived fresh frozen plasma compared with pooled, solvent detergent-treated plasma (Octaplas). AB - Adverse reactions to plasma transfusion are well documented. One of the most serious hazards of transfusion, transfusion-related acute lung injury (TRALI), has nearly as high an incidence of mortality and major morbidity as that reported for the transfusion of incorrect blood components. The specific mechanisms of plasma component-induced adverse reactions remain unclear, but a major contribution may be related to the presence of alloantibodies. In this study, a laboratory analysis was undertaken to determine the presence of HLA antibodies in leucodepleted single-donor-derived fresh frozen plasma (FFP). A comparison was made with a pooled plasma product that has undergone solvent detergent treatment. In total, 58 plasma samples from single-donor units of leucodepleted FFP were tested along with samples from 12 units (three for each ABO blood group) of the pooled plasma (Octaplas); Octapharma Ltd, Coventry, UK), for the presence of HLA antibodies. HLA-specific enzyme-linked immunosorbent assay (ELISA) methods were used to screen for their presence, and complement-dependent cytotoxicity and flow cytometry analyses were used to further define their presence, specificity and class [immunoglobulin G (IgG)/IgM]. In the study groups, HLA antibodies were found to be present in five of the single-donor units (9%) while the pooled plasma samples tested negative. PMID- 15548266 TI - Analysis of human leukocyte antigen (HLA)-G polymorphism in normal women and in women with recurrent spontaneous abortions. AB - Human leukocyte antigen (HLA)-G is a non-classical class I antigen. It has limited expression, but is high at the foetomaternal interface. This unique expression pattern of HLA-G suggests that it might be important for survival of the foetus during pregnancy. In the present study, 120 women with recurrent spontaneous abortions (RSA) and 120 fertile control women were genotyped for the HLA-G locus. This is the first report describing HLA-G polymorphism in normal fertile and RSA women from India. The allele HLA-G*010103 was higher in women with recurrent foetal losses. Interestingly, the HLA-G*010105 and G*010108 alleles were totally absent in normal fertile women but present in RSA women with frequencies of 1.7% and 0.4%, respectively. Allele G*010107 was absent in both the groups. The frequency of the null allele G*0105N was high (13.8%) in our population as compared to other world populations. Our data support the hypothesis that HLA-G polymorphism may contribute to recurrent foetal loss. PMID- 15548267 TI - Nomenclature for factors of the HLA system, update August 2004. PMID- 15548268 TI - Nomenclature for factors of the HLA system, update September 2004. PMID- 15548270 TI - Assessment of a contrast medium as an adjunct to endodontic radiography. AB - AIM: To assess if a contrast medium improved diagnostic yield of endodontic radiographs. METHODOLOGY: Forty-five extracted mandibular premolar teeth were radiographed in bucco-lingual and mesio-distal planes. Access cavities were prepared, pulpal tissue extirpated and Ultravist contrast medium introduced under pressure. Radiographs were retaken and the teeth cleared following perfusion with India ink. Three examiners assessed all the films for: number of roots, number of root canals, curvature of root and/or root canal, presence of lateral canals, presence of a single foramen or apical delta and the presence or absence of canal obstructions. The examiners' interpretations were compared with the anatomy revealed by clearing. RESULTS: Kappa scores were calculated for each of the examiners, for each set of radiographs, to assess the level of intra- and inter examiner agreement. Only moderate agreement was found throughout (kappa = 0.40 0.61). For multiple root canals a false-positive result was significantly more likely with contrast (P < 0.05). The use of contrast did not significantly improve the sensitivity of diagnosis of lateral canals or a single apical foramen. Contrast significantly increased the risk of falsely perceiving lateral canals (P < 0.002). Overall there was no statistically significant difference in the overall assessment of the anatomy of the root canals using contrast or plain radiographs (P > 0.2). CONCLUSIONS: Plain film radiographs confidently predict the presence of root or canal curvature but apical anatomy was assessed accurately in only 46% of cases. Plain radiographs were insensitive in assessing the number of root canals present, the presence of lateral canals and the occurrence of canal obstructions. The use of Ultravist contrast medium to improve diagnosis of root canal morphology of premolars is not supported. PMID- 15548271 TI - Reduction of connexin 43 expression in aged human dental pulp. AB - AIM: To investigate the expression of connexin 43 (CX43) mRNA in young and old human dental pulp tissues to determine the characteristics of CX43 expression. METHODOLOGY: Samples were obtained from human dental pulp of healthy young (17-23 years) and aged (>50 years) subjects. CX43 expression was determined by RT-PCR and by quantitative real-time RT-PCR (QRT-PCR). The threshold cycle (Ct) value, which reflects the amount of PCR, was calculated and the difference between value in the young pulp and that in the aged pulp was statistically analysed. RESULTS: RT-PCR analysis of human dental pulp tissue detected CX43 mRNA in all the samples. CX43 was abundantly expressed in young adult dental pulp, but expression of CX43 mRNA was dramatically decreased in aged human dental pulp. QRT-PCR analysis also showed the reduced expression of CX43 in aged pulp, and expression of CX43 in young pulp was significantly higher (about 10-fold, P < 0.01, Mann Whitney U-test). CONCLUSION: Reduction of CX43 expression may be associated with the loss of viability in human dental pulp, and is considered as one characteristic of aged pulp. PMID- 15548272 TI - Investigation of the effect of the coronal restoration quality on the composition of the root canal microflora in teeth with apical periodontitis by means of T RFLP analysis. AB - AIM: To investigate the effect of the radiographic and clinical quality of coronal restorations on the composition of the root canal flora of teeth with necrotic pulps and teeth with root fillings associated with apical periodontitis. METHODOLOGY: Twenty-eight necrotic pulps and 35 root filled canals with signs of apical periodontitis were studied. Both the coronal filling (presence of radiographically or clinically deficient margins and/or secondary caries) and the root filling (homogeneity and length) were scored. Bacterial root canal samples were taken with sterile paper points under rubber dam and using measures to prevent contamination. A DNA-based nonculture bacterial identification technique was used, namely terminal restriction fragment length polymorphism (T-RFLP) analysis. RESULTS: Twelve samples were negative for bacterial DNA. A total of 33 different terminal restriction fragments (TRFs) were detected. The Fusobacterium nucleatum/Streptococcus mitis group was the most frequently encountered TRF. The mean number of TRFs per necrotic pulp was 6.2 and 5.8 for the groups with acceptable and unacceptable coronal restorations, respectively. This difference was not significant. In the root filled group, these values (respectively, 5.2 and 8.6) were statistically significantly different (P < 0.05). The following parameters in root filled teeth had no significant influence on the mean numbers of TRFs detected: the length and homogeneity of the root filling and the type of tooth (anterior-premolar-molar). CONCLUSION: T-RFLP allowed the rapid assessment of bacterial biodiversity in root canal samples. The technique revealed the presence of bacteria that have rarely been described in the root canals of teeth with apical periodontitis. Biodiversity in the root filled group was high, as compared with culture-dependent studies where monoinfections were more frequently reported. Only in root filled teeth did defective coronal restorations have a statistically significant influence on the mean numbers of detected TRFs per sample. PMID- 15548273 TI - The effect of two different calcium hydroxide combinations on root dentine microhardness. AB - AIM: To evaluate the effect of a calcium hydroxide and glycerine mix and a calcium hydroxide and water mix on the microhardness of human root dentine. METHODOLOGY: Eleven freshly extracted maxillary canine and central incisor teeth were used. The teeth were sectioned transversally to produce a total of 22 dentine discs from the middle-third of the root. The specimens were divided into two groups of 11 discs each. Dentine samples were treated with either a Ca(OH)2 glycerine combination or a Ca(OH)2-distilled water combination for 1, 3 and 7 days. Dentine microhardness was measured with a Knoop indenter with a load of 100 g for 15 s before and during the experimental period. Each root disc received a series of three indentations around the pulp space, 1 mm from canal wall. RESULTS: Statistical analysis showed that both combinations significantly decreased dentine microhardness after 3 and 7 days (P < 0.01). The reduction in dentine microhardness following the use of a Ca(OH)2-glycerine combination was significantly greater than that after a Ca(OH)2-distilled water combination after 3 and 7 days (P < 0.01). CONCLUSION: The use of Ca(OH)2 combinations for intracanal dressing softens dentine. PMID- 15548274 TI - SEM evaluation of canal wall dentine following use of Mtwo and ProTaper NiTi rotary instruments. AB - AIM: To compare using scanning electron microscopy (SEM) root canal walls following instrumentation in vitro with two different rotary NiTi instruments. The hypothesis was that no difference should be observable between the experimental groups in terms of debris on canal walls and surface morphology. METHODOLOGY: Twenty-four single-rooted human teeth were selected. Two types of NiTi instruments were used, Mtwo (Sweden & Martina, Padova, Italy) and ProTaper (Dentsply Maillefer, Ballaigues, Switzerland). Irrigation for both groups was performed after each instrument change with 5% NaOCl, 3% H2O2 and 17% EDTA solutions. Three different areas (coronal, middle and apical thirds) of the root canal were evaluated using SEM. The canal wall of each sample was assessed and compared using a predefined scale of four parameters, namely, smear layer, pulpal debris, inorganic dentine debris, surface profile. Data were analysed statistically using the Kruskal-Wallis test (anova). RESULTS: A statistically significant difference (P < 0.01) was found between the apical third and the middle and coronal thirds for both groups. No difference was observable between instrumentation groups. In the apical third canal walls were often contaminated by inorganic debris and by smear layer. In the apical third, the surface profile was affected by uninstrumented regions, comprising dentine depressions and grooves in which predentine was still visible. CONCLUSION: Both instruments produced a clean and debris-free dentine surfaces in the coronal and middle thirds, but were unable to produce a dentine surfaces free from smear layer and debris in the apical third. The presence of deep grooves and depression on dentine walls in the apical third may well explain the presence of less instrumented areas. PMID- 15548275 TI - A Confocal Laser Scanning Microscope investigation of different dental adhesives bonded to root canal dentine. AB - AIM: To evaluate the resin-dentine interface of different adhesive systems and corresponding luting cements proposed for bonding fibre posts to root canal dentine. METHODOLOGY: Fifty extracted maxillary canines and central incisors were used. After root canal treatment the teeth were randomly divided into five groups of 10 teeth each. Fibre posts were inserted with five different adhesive systems and corresponding luting cements. Group 1: Clearfil Core/New Bond (Kuraray), group 2: Multilink (Vivadent), group 3: Panavia 21/ED Primer (Kuraray), group 4: PermaFlo DC (Ultradent), and group 5: Variolink II/Excite DSC (Vivadent). The primer was labelled in each case with 0.1% Rhodamine B isothiocyanate (RITC). Each root was sectioned into 2 mm thick slices at 1, 4 and 7 mm below the cementoenamel junction. The resin-dentine interface was evaluated using a Confocal Laser Scanning Microscope; the thickness of the hybrid layer and the number of resin tags were measured. The statistical analysis was performed using nonparametrical tests for comparisons between groups; for overall comparisons the Kruskal-Wallis test was used. Intraindividual analysis within teeth was performed using a linear model. RESULTS: The thickness (microm) of the hybrid layer of group 1 (5.45; SD 1.21), group 4 (3.36; SD 1.59), and group 5 (4.33; SD 1.19) was significantly higher than in the other groups (P < or = 0.05). The number of resin tags observed in group 1 was significantly higher than in groups 2-4 (P < 0.05), but did not differ from group 5. Each group showed significantly more resin tags in the coronal and in the central part of the root canal than in the apical part (P < 0.001). CONCLUSION: Conditioning of the root canal dentine with phosphoric acid and the use of one- and two-bottle-bonding systems gave a thicker and more uniform hybrid layer with considerably more resin tags than observed after the use of 'self-etching' adhesives. This might provide a more durable bond of the post to root canal dentine. PMID- 15548278 TI - Fine-scale spatial genetic correlation analyses reveal strong female philopatry within a brush-tailed rock-wallaby colony in southeast Queensland. AB - We combine spatial data on home ranges of individuals and microsatellite markers to examine patterns of fine-scale spatial genetic structure and dispersal within a brush-tailed rock-wallaby (Petrogale penicillata) colony at Hurdle Creek Valley, Queensland. Brush-tailed rock-wallabies were once abundant and widespread throughout the rocky terrain of southeastern Australia; however, populations are nearly extinct in the south of their range and in decline elsewhere. We use pairwise relatedness measures and a recent multilocus spatial autocorrelation analysis to test the hypotheses that in this species, within-colony dispersal is male-biased and that female philopatry results in spatial clusters of related females within the colony. We provide clear evidence for strong female philopatry and male-biased dispersal within this rock-wallaby colony. There was a strong, significant negative correlation between pairwise relatedness and geographical distance of individual females along only 800 m of cliff line. Spatial genetic autocorrelation analyses showed significant positive correlation for females in close proximity to each other and revealed a genetic neighbourhood size of only 600 m for females. Our study is the first to report on the fine-scale spatial genetic structure within a rock-wallaby colony and we provide the first robust evidence for strong female philopatry and spatial clustering of related females within this taxon. We discuss the ecological and conservation implications of our findings for rock-wallabies, as well as the importance of fine-scale spatial genetic patterns in studies of dispersal behaviour. PMID- 15548276 TI - An analysis of endodontic treatment with three nickel-titanium rotary root canal preparation techniques. AB - AIM: To investigate clinical results of root canal treatment performed with the aid of nickel-titanium (NiTi) rotary instruments. SUMMARY: A total of 179 patients underwent root canal treatment with either (A) Lightspeed, or (B) ProFile .04 or (C) ProFile .04 and .06 or GT rotary instruments to create tapered preparations. In groups A and B, laterally condensed gutta-percha and AH Plus were used. Canals in group C were obturated with System B, Obtura II and Roth's 801 sealer. Initial and recall radiographs were assessed using the periapical index (PAI). Outcomes were analysed using chi-square tests, event-time analyses and logistic regression models. Two hundred and thirty-three teeth were radiographically assessed after a mean interval of 25.4 +/- 11.8 months. Favourable outcome of treatment, defined as PAI < 3 at recall was 86.7%. Logistic regression analysis and univariate analyses indicated that teeth with preoperative PAI scores >2 and retreated teeth had a significantly lower chance of healing compared with periapically healthy teeth and primary treatments, respectively. Preparation technique, length of fill and the type of sealer did not significantly affect healing rates. KEY LEARNING POINTS: Root canal treatment with NiTi root canal instrumentation systems renders favourable outcomes in more than 86% of the cases. Outcome is significantly affected by preoperative diagnoses but not by the specific choice of instrumentation system. PMID- 15548279 TI - Mitochondrial DNA differentiation between two forms of trout Salmo letnica, endemic to the Balkan Lake Ohrid, reflects their reproductive isolation. AB - Mitochondrial haplotype diversity in sympatric populations of Ohrid trout, Salmo letnica was investigated by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mtDNA control region and ND1, ND3/4, ND5/6 segments. A 310 bp fragment at the 5' end, and a 340-572 bp fragment at the 3' end of the control region were sequenced from representatives of the populations studied. Based on pairwise comparison of the sequences, five new haplotypes were identified plus one identical with the brown trout Andalusian haplotype from the southern Iberian Peninsula. The combination of both RFLP and sequence data sets yielded a total of 10 composite haplotypes. A high degree of genetic subdivision between S. letnica typicus and S. letnica aestivalis populations was observed. The notion of a sympatric origin for the two morphs is discussed. Length variation of the mtDNA control region due to the presence of an 82 bp unit, tandemly repeated one to four times, in the region between the conserved sequence block-3 (CSB-3) and the gene for phenylalanine tRNA is reported. Further, we demonstrate that a single duplication of the approximately 82 bp repeat unit is a common element of the salmonid mitochondrial control region. The unique genetic structure of Ohrid trout represents a highly valuable genetic resource that deserves appropriate management and conservation. PMID- 15548280 TI - Spatial genetic structure in a metapopulation of the land snail Cepaea nemoralis (Gastropoda: Helicidae). AB - Habitat fragmentation is a major force affecting demography and genetic structure of wild populations, especially in agricultural landscapes. The land snail Cepaea nemoralis (L.) was selected to investigate the impact of habitat fragmentation on the spatial genetic structure of an organism with limited dispersal ability. Genetic and morphological patterns were investigated at a local scale of a 500 m transect and a mesoscale of 4 x 4 km in a fragmented agricultural landscape while accounting for variation in the landscape using least-cost models. Analysis of microsatellite loci using expected heterozygosity (HE), pairwise genetic distance (FST/1-FST) and spatial autocorrelograms (Moran's I) as well as shell characteristics revealed spatial structuring at both scales and provided evidence for a metapopulation structure. Genetic diversity was related to morphological diversity regardless of landscape properties. This pointed to bottlenecks caused by founder effects after (re)colonization. Our study suggests that metapopulation structure depended on both landscape features and the shape of the dispersal function. A range of genetic spatial autocorrelation up to 80 m at the local scale and up to 800 m at the mesoscale indicated leptokurtic dispersal patterns. The metapopulation dynamics of C. nemoralis resulted in a patchwork of interconnected, spatially structured subpopulations. They were shaped by gene flow which was affected by landscape features, the dispersal function and an increasing role of genetic drift with distance. PMID- 15548281 TI - Population genetics and phylogeography of endangered Oxytropis campestris var. chartacea and relatives: arctic-alpine disjuncts in eastern North America. AB - Fassett's locoweed (Oxytropis campestris var. chartacea, Fabaceae) is an endangered perennial endemic to Wisconsin. Patterns of genetic variation within and among six remaining populations and their relationship to other members of the O. campestris complex were analysed using AFLPs from 140 accessions across northern North America. Within-population measures of genetic diversity were high (mean expected heterozygosity HE = 0.16; mean nucleotide diversity pi = 0.015) compared with other herbaceous plants. Estimates of among-population differentiation were low (FST = 0.12; PhiST = 0.29), consistent with outcrossing. Genetic and geographical distances between populations were significantly correlated within Fassett's locoweed (r2 = 0.73, P < 0.002 for Mantel test) and O. campestris as a whole (r2 = 0.63, P < 0.0001). Individual and population-based phylogenetic analyses showed that Fassett's locoweed is monophyletic and sister to O. campestris var. johannensis. Morphometric analyses revealed significant differences between Fassett's locoweed and populations of var. johannensis. The first chromosome count for Fassett's locoweed indicates that it is tetraploid (2n = 32), unlike hexaploid var. johannensis. High within-population diversity and relatively low among-population differentiation are consistent with populations of Fassett's locoweed being relicts of a more continuous Pleistocene distribution. Our data support the continued recognition of Fassett's locoweed and protection under federal and state regulations. High levels of genetic diversity within populations suggest that maintain-ing the ecological conditions that favour the life cycle of this plant may be a more pressing concern than the erosion of genetic variation. PMID- 15548282 TI - Ecological and life history characteristics predict population genetic divergence of two salmonids in the same landscape. AB - Ecological and life history characteristics such as population size, dispersal pattern, and mating system mediate the influence of genetic drift and gene flow on population subdivision. Bull trout (Salvelinus confluentus) and mountain whitefish (Prosopium williamsoni) differ markedly in spawning location, population size and mating system. Based on these differences, we predicted that bull trout would have reduced genetic variation within and greater differentiation among populations compared with mountain whitefish. To test this hypothesis, we used microsatellite markers to determine patterns of genetic divergence for each species in the Clark Fork River, Montana, USA. As predicted, bull trout had a much greater proportion of genetic variation partitioned among populations than mountain whitefish. Among all sites, FST was seven times greater for bull trout (FST = 0.304 for bull trout, 0.042 for mountain whitefish. After removing genetically differentiated high mountain lake sites for each species FST, was 10 times greater for bull trout (FST = 0.176 for bull trout; FST = 0.018 for mountain whitefish). The same characteristics that affect dispersal patterns in these species also lead to predictions about the amount and scale of adaptive divergence among populations. We provide a theoretical framework that incorporates variation in ecological and life history factors, neutral divergence, and adaptive divergence to interpret how neutral and adaptive divergence might be correlates of ecological and life history factors. PMID- 15548283 TI - Impacts of gene flow and logging history on the local genetic structure of a scattered tree species, Sorbus torminalis L. Crantz. AB - Sorbus torminalis L. Crantz is a colonizing tree species usually found at low density in managed European forests. Using six microsatellite markers, we investigated spatial and temporal patterns of genetic structure within a 472-ha population of 185 individuals to infer processes shaping the distribution of genetic diversity. Only eight young stems were found to be the result of vegetative reproduction. Despite high levels of gene flow (standard deviation of gene dispersal = 360 m), marked patterns of isolation by distance were detected, associated with an aggregated distribution of individuals in approximately 100-m patches. This spatial structure of both genes and individuals is likely to result from patterns of seedling recruitment combined with low tree density. Our results suggest that landscape factors and logging cycles markedly shape the distribution of favourable sites for seedling establishment, which are then colonized by sibling cohorts as a result of joint seed transportation by frugivores. These combined genetic and demographic processes result in similar genetic structure both within and among logging units. However, conversion to high forest may enhance genetic structuring. PMID- 15548284 TI - Mating frequency, within-colony relatedness and male production in a yellow jacket wasp, Dolichovespula arenaria. AB - We examined the mating frequencies of queens in a social wasp, Dolichovespula arenaria (Vespinae) using DNA microsatellites. Five of the seven colonies supported the hypothesis of single mating by queens. The other two colonies supported two and three matings, with effective paternity of 1.48 and 1.91. Mean worker relatedness was 0.77 +/- 0.09. In two of the four male-containing colonies, all were likely progeny of the queen. In the other two colonies workers produced 8 and 14% of the male wasps. Overall, 94.3% of the male wasps were likely progeny of the queen. These patterns are consistent with published studies of vespine wasps. PMID- 15548285 TI - Genetic drift outweighs balancing selection in shaping post-bottleneck major histocompatibility complex variation in New Zealand robins (Petroicidae). AB - The Chatham Island black robin, Petroica traversi, is a highly inbred, endangered passerine with extremely low levels of variation at hypervariable neutral DNA markers. In this study we investigated variation in major histocompatibility complex (MHC) class II genes in both the black robin and its nonendangered relative, the South Island robin Petroica australis australis. Previous studies have shown that Petroica have at least four expressed class II B MHC genes. In this study, the sequences of introns flanking exon 2 of these loci were characterized to design primers for peptide-binding region (PBR) sequence analysis. Intron sequences were comprised of varying numbers of repeated units, with highly conserved regions immediately flanking exon 2. Polymerase chain reaction primers designed to this region amplified three or four sequences per black robin individual, and eight to 14 sequences per South Island robin individual. MHC genes are fitness-related genes thought to be under balancing selection, so they may be more likely to retain variation in bottlenecked populations. To test this, we compared MHC variation in the black robin with artificially bottlenecked populations of South Island robin, and with their respective source populations, using restriction fragment length polymorphism analyses and DNA sequencing of the PBR. Our results indicate that the black robin is monomorphic at class II B MHC loci, while both source and bottlenecked populations of South Island robin have retained moderate levels of variation. Comparison of MHC variation with minisatellite DNA variation indicates that genetic drift outweighs balancing selection in determining MHC diversity in the bottlenecked populations. However, balancing selection appears to influence MHC diversity over evolutionary timescales, and the effects of gene conversion are evident. PMID- 15548286 TI - Phylogeography of Bufo fowleri at its northern range limit. AB - Many of the species that recolonized previously glaciated areas in the Great Lakes basin of North America over the past 10-12,000 years exhibit genetic evidence of multiple invasion routes and present-day secondary contact between deeply divergent lineages. With this in mind, we investigated the phylogeographical structure of genetic variability in Fowler's toads (Bufo fowleri) at the northern edge of its distribution where its range encircles the Lake Erie basin. Because B. fowleri is so closely tied to habitats along the Lake Erie shoreline, we would expect to find clear evidence of the number of invasions leading to the species' colonization of the northern shore. A 540 bp sequence from the mitochondrial control region was amplified and analysed for 158 individuals from 21 populations. Interpopulation sequence variation ranged from 0% to 6%. Phylogenetic analysis of p-distance using the neighbor-joining method revealed two deeply divergent (6% sequence divergence) mtDNA lineages (Phylogroup 1 and 2), possibly arising as a result of secondary contact of populations that entered the region from two separate glacial refugia. However, the phylogeographical pattern was not simple. The populations at Long Point, on the north shore of Lake Erie, clustered with the population from Indiana Dunes on Lake Michigan to form Phylogroup 2 whereas all other B. fowleri populations examined from both sides of Lake Erie constituted Phylogroup 1. Furthermore, mtDNA sequences from the related species Bufo americanus, obtained from populations outside the range of B. fowleri, clustered with mtDNA haplotypes of B. fowleri Phylogroup 1, indicating the possibility of partial introgression of mitochondria from one species to the other. PMID- 15548287 TI - Hybrid zone structure and the potential role of selection in hybridizing populations of native westslope cutthroat trout (Oncorhynchus clarki lewisi) and introduced rainbow trout (O. mykiss). AB - Introgressive hybridization is a common feature of many zones of contact between divergent lineages of fishes. This is particularly common when taxa that are normally allopatric come into artificial (human-induced) secondary contact. We examined 18 native populations of westslope cutthroat trout (Oncorhynchus clarki lewisi, WCT) to determine the extent of introgressive hybridization with introduced rainbow trout (O. mykiss, RBT) and the genetic structure of hybridizing populations in the upper Kootenay River, southeastern British Columbia, Canada. Using four diagnostic nuclear loci we calculated a hybrid index, inbreeding coefficient, FIS, and the linkage disquilibrium correlation coefficient, Rij, for each locality to determine the distribution of genotypes in each population. We also categorized the 142 hybrid individuals found across localities into four hybrid classes based on their genotypes. The majority of localities (11/18) showed a unimodal distribution of genotypes skewed towards genotypes of WCT. Two localities, however (lower Gold Creek and Lodgepole Creek) showed a flat to bimodal distribution and one site (lower Bull River) showed a unimodal distribution skewed towards RBT genotypes. The majority of hybrid individuals were classified genotypically as WCT backcrosses (59%) and post-F1 individuals (24%). We found a skewed ratio of pure WCT to pure RBT (17:1) and only four F1 hybrids (3%), suggesting that the spread of RBT alleles may be facilitated by hybrids straying to neighbouring populations. We also tested for the action of selection in one population using cohort analyses, but found little evidence of differential selection between pure WCT and hybrid individuals. Pooled across age classes there were significant differences in genotypic frequencies among loci suggesting differential introgression. There was no asymmetry to the hybridization between rainbow trout and westslope cutthroat trout because both species' mitochondrial DNA haplotypes were observed at similar frequencies in the hybrids. Our analyses suggest that hybrid swarms are likely to form in the upper Kootenay River drainage and that certain native WCT populations in British Columbia are at risk of local genomic extinction. PMID- 15548288 TI - The rare silver gum, Eucalyptus cordata, is leaving its trace in the organellar gene pool of Eucalyptus globulus. AB - The process of genetic assimilation of rare species by hybridizing congeners has been documented in a number of plant genera. This raises the possibility that some of the genetic diversity found in phylogeographical studies of widespread species has been acquired through hybridization with species that are now rare or extinct. In this fine-scale phylogeographical analysis, we show that a rare eucalypt species is leaving its trace in the chloroplast genome of a more abundant congener. The heart-leafed silver gum, Eucalyptus cordata, is a rare endemic of south-eastern Tasmania. Its populations are scattered amidst populations of more abundant related species, including the Tasmanian blue gum, Eucalyptus globulus. Using 339 samples from across the full range of both species, we compared chloroplast (cp) DNA haplotype phylogeography in E. globulus and E. cordata. The genealogy and distribution of chloroplast haplotypes suggest that E. globulus has acquired cpDNA from E. cordata in at least four different mixed populations. Shared haplotypes are highest in E. globulus sampled within 2 km of known E. cordata populations and drop to zero at a distance of 25 km from the nearest known E. cordata population. Localized haplotype sharing occurs in the absence of obvious hybrid zones or locally shared nuclear ribosomal DNA sequences. Given that the future loss of E. cordata from some mixed populations is likely, these findings indicate that phylogeographical analyses of organellar DNA should consider the possibility of introgression, even from species that have been eliminated from the sites of interest. PMID- 15548289 TI - New evidence for parallel evolution of colour patterns in Malagasy poison frogs (Mantella). AB - Malagasy poison frogs of the genus Mantella are diurnal and toxic amphibians of highly variable and largely aposematic coloration. Previous studies provided evidence for several instances of homoplastic colour evolution in this genus but were unable to sufficiently resolve relationships among major species groups or to clarify the phylogenetic position of several crucial taxa. Here, we provide cytochrome b data for 143 individuals of three species in the Mantella madagascariensis group, including four newly discovered populations. Three of these new populations are characterized by highly variable coloration and patterns but showed no conspicuous increase of haplotype diversity which would be expected under a scenario of secondary hybridization or admixture of chromatically uniform populations. Several populations of these variable forms and of M. crocea were geographically interspersed between the distribution areas of Mantella aurantiaca and Mantella milotympanum. This provides further support for the hypothesis that the largely similar uniformly orange colour of the last two species evolved in parallel. Phylogenies based on over 2000 bp of two nuclear genes (Rag-1 and Rag-2) identified reliably a clade of the Mantella betsileo and Mantella laevigata groups as sister lineage to the M. madagascariensis group, but did not support species within the latter group as monophyletic. The evolutionary history of these frogs might have been characterized by fast and recurrent evolution of colour patterns, possibly triggered by strong selection pressures and mimicry effects, being too complex to be represented by simple bifurcating models of phylogenetic reconstruction. PMID- 15548290 TI - Long-distance gene flow and cross-Andean dispersal of lowland rainforest bees (Apidae: Euglossini) revealed by comparative mitochondrial DNA phylogeography. AB - Euglossine bees (Apidae; Euglossini) exclusively pollinate hundreds of orchid species and comprise up to 25% of bee species richness in neotropical rainforests. As one of the first studies of comparative phylogeography in a neotropical insect group, we performed a mitochondrial DNA (mtDNA)-based analysis of 14 euglossine species represented by populations sampled across the Andes and/or across the Amazon basin. The mtDNA divergences within species were consistently low; across the 12 monophyletic species the mean intraspecific divergence among haplotypes was 0.9% (range of means, 0-1.9%). The cytochrome oxidase 1 (CO1) divergence among populations separated by the Andes (N = 11 species) averaged 1.1% (range 0.0-2.0%). The mtDNA CO1 data set displayed homogeneous rates of nucleotide substitution, permitting us to infer dispersal across the cordillera long after the final Andean uplift based on arthropod molecular clocks of 1.2-1.5% divergence per million years. Gene flow across the 3000-km breadth of the Amazon basin was inferred from identical cross-Amazon haplotypes found in five species. Although mtDNA haplotypes for 12 of the 14 euglossine species were monophyletic, a reticulate CO1 phylogeny was recovered in Euglossa cognata and E. mixta, suggesting large ancestral populations and recent speciation. Reference to closely related outgroups suggested recent speciation for the majority of species. Phylogeographical structure across a broad spatial scale is weaker in euglossine bees than in any neotropical group previously examined, and may derive from a combination of Quaternary speciation, population expansion and/or long-distance gene flow. PMID- 15548291 TI - Testing geographical pathways of speciation in a recent island radiation. AB - Determining the mode, or geographical context, of speciation is a critical first step to understanding the evolutionary mechanisms that cause new species to arise. In this study, we estimated phylogenetic relationships in the cerasina species group of the Hawaiian cricket genus Laupala (Orthoptera: Gryllidae) to test competing phylogeographical hypotheses and thus infer the mode of speciation. A previous phylogenetic result based on nuclear sequence data suggested that populations of L. cerasina on the Big Island of Hawaii are the result of two independent colonizations from Maui, implying parallel speciation and convergent song evolution, and contradicting systematic hypotheses based on behavioural and morphological data. We used amplified fragment length polymorphisms to investigate further the relationships among species and populations in the cerasina species group. Results of these analyses provide a robust estimate of phylogenetic relationships and support the phylogeographical history indicated by behavioural and morphological data. PMID- 15548292 TI - Natal homing in juvenile loggerhead turtles (Caretta caretta). AB - Juvenile loggerhead turtles (Caretta caretta) from West Atlantic nesting beaches occupy oceanic (pelagic) habitats in the eastern Atlantic and Mediterranean, whereas larger juvenile turtles occupy shallow (neritic) habitats along the continental coastline of North America. Hence the switch from oceanic to neritic stage can involve a trans-oceanic migration. Several researchers have suggested that at the end of the oceanic phase, juveniles are homing to feeding habitats in the vicinity of their natal rookery. To test the hypothesis of juvenile homing behaviour, we surveyed 10 juvenile feeding zones across the eastern USA with mitochondrial DNA control region sequences (N = 1437) and compared these samples to potential source (nesting) populations in the Atlantic Ocean and Mediterranean Sea (N = 465). The results indicated a shallow, but significant, population structure of neritic juveniles (PhiST = 0.0088, P = 0.016), and haplotype frequency differences were significantly correlated between coastal feeding populations and adjacent nesting populations (Mantel test R2 = 0.52, P = 0.001). Mixed stock analyses (using a Bayesian algorithm) indicated that juveniles occurred at elevated frequency in the vicinity of their natal rookery. Hence, all lines of evidence supported the hypothesis of juvenile homing in loggerhead turtles. While not as precise as the homing of breeding adults, this behaviour nonetheless places juvenile turtles in the vicinity of their natal nesting colonies. Some of the coastal hazards that affect declining nesting populations may also affect the next generation of turtles feeding in nearby habitats. PMID- 15548293 TI - Divergence in gene expression related to variation in host specificity of an ectomycorrhizal fungus. AB - Ectomycorrhizae are formed by mutualistic interactions between fungi and the roots of woody plants. During symbiosis the two organisms exchange carbon and nutrients in a specific tissue that is formed at the contact between a compatible fungus and plant. There is considerable variation in the degree of host specificity among species and strains of ectomycorrhizal fungi. In this study, we have for the first time shown that this variation is associated with quantitative differences in gene expression, and with divergence in nucleotide sequences of symbiosis-regulated genes. Gene expression and sequence evolution were compared in different strains of the ectomycorrhizal fungus Paxillus involutus; the strains included Nau, which is not compatible with birch and poplar, and the two compatible strains Maj and ATCC200175. On a genomic level, Nau and Maj were very similar. The sequence identity was 98.9% in the 16 loci analysed, and only three out of 1075 genes analysed by microarray-based hybridizations had signals indicating differences in gene copy numbers. In contrast, 66 out of the 1075 genes were differentially expressed in Maj compared to Nau after contact with birch roots. Thirty-seven of these symbiosis-regulated genes were also differentially expressed in the ATCC strain. Comparative analysis of DNA sequences of the symbiosis-regulated genes in different strains showed that two of them have evolved at an enhanced rate in Nau. The sequence divergence can be explained by a decreased selection pressure, which in turn is determined by lower functional constraints on these proteins in Nau as compared to the compatible strains. PMID- 15548294 TI - Major histocompatibility complex variation and mate choice in a lekking bird, the great snipe (Gallinago media). AB - Genes of the major histocompatibility complex (MHC) play a major part in the activation of the vertebrate immune system. In addition, they also appear to function as cues for mate choice. In mammals especially, several kinds of MHC dependent mate choice have been hypothesized and observed. These include choice of mates that share no or few alleles with the choosing individual, choice of mates with alleles that differ as much as possible from the choosing individual, choice of heterozygous mates, choice of certain genotypes and choice of rare alleles. We investigated these different aspects of mate choice in relation to MHC in a lekking bird species, the great snipe (Gallinago media). We found no evidence for MHC disassortative mating, no preference for males with many MHC alleles and no preference for rare alleles. However, we did find that some allelic lineages were more often found in males with mating success than in males without mating success. Females do not seem to use themselves as references for the MHC-dependent mate choice, rather they seem to prefer males with certain allele types. We speculate that these alleles may be linked to resistance to common parasites. PMID- 15548295 TI - Prevalence and differential host-specificity of two avian blood parasite genera in the Australo-Papuan region. AB - The degree to which widespread avian blood parasites in the genera Plasmodium and Haemoproteus pose a threat to novel hosts depends in part on the degree to which they are constrained to a particular host or host family. We examined the host distribution and host-specificity of these parasites in birds from two relatively understudied and isolated locations: Australia and Papua New Guinea. Using polymerase chain reaction (PCR), we detected infection in 69 of 105 species, representing 44% of individuals surveyed (n = 428). Across host families, prevalence of Haemoproteus ranged from 13% (Acanthizidae) to 56% (Petroicidae) while prevalence of Plasmodium ranged from 3% (Petroicidae) to 47% (Ptilonorhynchidae). We recovered 78 unique mitochondrial lineages from 155 sequences. Related lineages of Haemoproteus were more likely to derive from the same host family than predicted by chance at shallow (average LogDet genetic distance = 0, n = 12, P = 0.001) and greater depths (average distance = 0.014, n = 11, P < 0.001) within the parasite phylogeny. Within two major Haemoproteus subclades identified in a maximum likelihood phylogeny, host-specificity was evident up to parasite genetic distances of 0.029 and 0.007 based on logistic regression. We found no significant host relationship among lineages of Plasmodium by any method of analysis. These results support previous evidence of strong host-family specificity in Haemoproteus and suggest that lineages of Plasmodium are more likely to form evolutionarily-stable associations with novel hosts. PMID- 15548296 TI - On the origins of medfly invasion and expansion in Australia. AB - As a result of their rapid expansion and large larval host range, true fruit flies are among the world's most important agricultural pest species. Among them, Ceratitis capitata has become a model organism for studies on colonization and invasion processes. The genetic aspects of the medfly invasion process have already been analysed throughout its range, with the exception of Australia. Bioinvasion into Australia is an old event: medfly were first captured in Australia in 1895, near Perth. After briefly appearing in Tasmania and the eastern states of mainland Australia, medfly had disappeared from these areas by the 1940s. Currently, they are confined to the western coastal region. South Australia seems to be protected from medfly infestations both by the presence of an inhospitable barrier separating it from the west and by the limited number of transport routes. However, numerous medfly outbreaks have occurred since 1946, mainly near Adelaide. Allele frequency data at 10 simple sequence repeat loci were used to study the genetic structure of Australian medflies, to infer the historical pattern of invasion and the origin of the recent outbreaks. The combination of phylogeographical analysis and Bayesian tests showed that colonization of Australia was a secondary colonization event from the Mediterranean basin and that Australian medflies were unlikely to be the source for the initial Hawaiian invasion. Within Australia, the Perth area acted as the core range and was the source for medfly bioinvasion in both Western and South Australia. Incipient differentiation, as a result of habitat fragmentation, was detected in some localized areas at the periphery of the core range. PMID- 15548297 TI - Distribution of genetic variation in the growth hormone 1 gene in Atlantic salmon (Salmo salar) populations from Europe and North America. AB - The level and hierarchical distribution of genetic variation in complete sequences of the Atlantic salmon (Salmo salar) growth hormone (GH1) gene were investigated in populations from Europe and North America with a view to inferring the major evolutionary forces affecting genetic variation at this locus. Seventeen polymorphic sites were identified in complete sequences from nine populations, with levels of noncoding (intron and untranslated region sequences) nucleotide diversity being similar to those observed in other species. No variation, however, was observed in exonic sequences, indicating that nucleotide diversity in the Atlantic salmon GH1 gene is three and 25 times less than that estimated for human and Drosophila coding sequences, respectively. This suggests that purifying selection is the predominant contemporary force controlling the molecular evolution of GH1 coding sequences. Comparison of haplotype relationships within and between populations indicated that differentiation between populations from Europe and North America was greater than within-continent comparisons. However, several haplotypes observed in the northernmost European populations were more similar to those observed in North American than to any other haplotypes observed in Europe. This is most likely to be a result of historical, rather than contemporary, gene flow. Neutrality test statistics, such as Tajima's D, were significantly positive in the European populations in which North American-like haplotypes were observed. Although a positive Tajima's D is commonly interpreted as the signal of balancing selection, a more likely explanation in this case is that either historical migration or ascertainment bias, rather than within population local adaptation, has given rise to an excess of intermediate frequency alleles. PMID- 15548298 TI - Quantitative analysis of amoA mRNA expression as a new biomarker of ammonia oxidation activities in a complex microbial community. AB - AIMS: To quantitatively analyse the changes to amoA mRNA (ammonia mono-oxygenase encoding mRNA) profiles in response to a change in ammonia oxidation activity in a complex microbial community. METHODS AND RESULTS: The amoA mRNA levels in both a batch-mode incubation and a continuously fed nitrification reactor were determined by real-time reverse transcription-PCR analysis. The amoA mRNA level changed rapidly in response to the change in environmental conditions which affect ammonia oxidation activity. CONCLUSION: An increase in amoA mRNA level can be detected within 1-2 h in response to an initiation of cell activity whereas a decrease in amoA mRNA level is detected within 24 h in response to a cessation of activity. SIGNIFICANCE AND IMPACT OF THE STUDY: amoA mRNA, which shows sensitive response to ammonia oxidation activity, can be used as a biomarker of ammonia oxidation activity in wastewater treatment processes where many bacterial species exist. PMID- 15548299 TI - Glycopeptide-resistance transferability from vancomycin-resistant enterococci of human and animal source to Listeria spp. AB - AIMS: The glycopeptide-resistance transferability from vancomycin-resistant enterococci (VRE) of clinical and animal origin to different species of Listeria was investigated. METHODS AND RESULTS: Of 36 matings, performed on membrane filter, the glycopeptide resistance was successfully transferred in six attempts, five with donors of animal origin and only one with donors from clinical source. The acquired glycopeptide resistance in Listeria transconjugants was confirmed by the presence of the conjugative plasmid band and by the amplification of the 732 bp fragment of vanA gene in transferred plasmids. CONCLUSIONS: Despite the lower number of bacteria used in this study, the source of enterococci influenced the outcome of mating. Moreover transferred VanA plasmid induced a different expression in Listeria transconjugants, suggesting that gene expression might be influenced by species affiliation of recipients. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data strengthen the opinion that enterococci are an important source of resistance genes for Listeria via the transfer of movable genetic elements. As these strains are commonly found in the same habitats, a horizontal spread of glycopeptide resistance in Listeria spp. could be possible. PMID- 15548300 TI - Comparison of three immunological methods for detecting staphylococcal enterotoxins from food. AB - AIMS: Immunologically based assays for the detection of staphylococcal enterotoxins are numerous. These techniques include radio immunosorbent assays and enzyme-linked immunosorbent assays (ELISA), some of which are available as commercial kits. The purpose of this study was to compare the performances of three commercial immunoassays. METHODS AND RESULTS: Two automated detection systems, VIDAS SET bioMerieux, VIDAS SET2 bioMerieux and an ELISA method, TRANSIA PLATE Staphylococcal Enterotoxins Diffchamb were compared for detecting different quantities of purified staphylococcal enterotoxins (A, B, C2, D and E) added to food. CONCLUSIONS: VIDAS SET2 had a greater specificity (100%) and sensitivity than VIDAS SET and TRANSIA PLATE Staphylococcal Enterotoxins. More precisely, VIDAS SET2 could detect <0.5 ng g(-1) of toxins A and B, <1 ng g(-1) of toxins C2 and E and 1 ng g(-1) of toxins D and E. SIGNIFICANCE AND IMPACT OF THE STUDY: Because staphylococcal food poisoning (resulting from ingestion of low levels of staphylococcal enterotoxins) is one of the most common forms of foodborne illness there is a need for specific and sensitive methods for detecting these enterotoxins. VIDAS SET2 appears to be suitable for detecting staphylococcal enterotoxins from food. PMID- 15548301 TI - Isolation and characterization of sulphur-oxidizing Thiomonas sp. and its potential application in biological deodorization. AB - AIMS: To isolate and characterize a sulphur-oxidizing bacterial strain from activated sludge and to evaluate its potential application in biological deodorization. METHODS AND RESULTS: A dominant sulphur-oxidizing bacterial strain, designated as strain SS, was isolated from an enrichment culture using thiosulphate as a sole energy source and CO2 as a sole carbon source. The cells of this organism were aerobic, rod-shaped, Gram-negative and motile. Strain SS could grow autotrophically, heterotrophically as well as mixotrophically. Autotrophic growth was observed at pH values ranging from 2.3 to 9.0. Phylogenetic analyses revealed that strain SS belonged to Group 1 of the genus Thiomonas, closely related to Thiomonas perometabolis and Thiomonas intermedia. The thiosulphate oxidation rates of strain SS at different pH values were evaluated in terms of oxygen uptake using a Micro-Oxymax respirometer. The results showed that the maximum oxidation rate of 5.65 mg l(-1) h(-1) occurred at 56 h of growth and pH 6.0. Continuous H2S removal study demonstrated that strain SS could remove more than 99% of H2S when the inlet concentration was below 58.6 ppm. Further increase of the inlet concentration to 118 ppm gave rise to a decline in the removal efficiency to ca 90%. CONCLUSIONS: The strong acidification of the culture medium during the later period could result in the deterioration of the growth activity and the metabolism activity of strain SS. In practical application, the problems caused by the end-product inhibition and the acidification can be alleviated by periodical replacement of culture medium with fresh medium. Given the physiological flexibility and the ability to remove H2S rapidly and efficiently, strain SS could be a good 'deodorizing' candidate. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that Thiomonas species has been reported for biological deodorization application. PMID- 15548302 TI - Limitations of membrane cultures as a model solid-state fermentation system. AB - AIMS: To examine the reliability of membrane cultures as a model solid-state fermentation (SSF) system. METHODS AND RESULTS: In overcultures of Aspergillus oryzae on sterilized wheat flour discs overlaid with a polycarbonate membrane, we demonstrated that the presence of membrane filters reduced the maximum respiration rate (up to 50%), and biomass and alpha-amylase production. We also show that the advantage of membrane cultures, i.e. total recovery of biomass, is not very evident for the system used, while the changes in metabolism and kinetics are serious drawbacks. CONCLUSIONS: The use of membrane cultures is artificial and without substantial benefits and therefore has to be carefully considered. SIGNIFICANCE AND IMPACT OF THE STUDY: In future studies on kinetics and stoichiometry of SSF, one should not completely rely on experiments using membrane cultures as a model SSF system. PMID- 15548303 TI - Evaluation of two real-time polymerase chain reaction pathogen detection kits for Salmonella spp. in food. AB - AIMS: To evaluate the LightCycler Salmonella Detection Kit and the TaqMan Salmonella Gold Detection and Quantitation Kit for the real-time PCR detection of Salmonella in various food samples. METHODS AND RESULTS: Ready-to-eat foods and raw food samples were artificially contaminated with Salmonella serotypes. In the specificity test, bacterial DNA extracted from sample pre-enrichment culture was analysed with the detection kits performed respectively on the LightCycler Instrument or the ABI Prism 7000 Sequence Detection System. No false-positive or false-negative results were obtained, although the LightCycler system generated invalid PCR results on two occasions. In the sensitivity test using the LightCycler system, Salmonella could be detected in pre-enrichment cultures of 25 g samples inoculated with as low as 1.5 x 10(3) CFU (depending on food type), and false-negative results were obtained for samples with low inoculum levels. CONCLUSIONS: Two commercial kits for real-time PCR detection of Salmonella were evaluated. SIGNIFICANCE AND IMPACT OF THE STUDY: Evaluation using more food types and matrices, and foods that contain low number of Salmonella or high number of other competing bacteria, is needed before adopting the real-time PCR technique for routine food tests. PMID- 15548304 TI - Comparison of fluorogenic and chromogenic assay systems in the detection of Escherichia coli O157 by a novel polymyxin-based ELISA. AB - AIMS: Different indicator enzymes and fluorogenic or chromogenic substrates were compared as detector systems in a novel polymyxin-based enzyme-linked immunosorbent assay (ELISA) for Escherichia coli O157 lipopolysaccharide (LPS) antigens. METHODS AND RESULTS: An ELISA system was developed using polymyxin immobilized in the wells of a microtitre plate as a high-affinity adsorbent for E. coli O157 LPS antigens, which were immunoenzymatically detected using anti-E. coli O157 antibody-enzyme conjugates. With peroxidase as the indicator enzyme the fluorogenic substrates Amplex Red and QuantaBlu produced only slight improvement in the performance characteristics of the polymyxin-ELISA compared with the use of the chromogenic substrate tetramethylbenzidine (TMB). On the other hand, with alkaline phosphatase as the indicator enzyme a pronounced improvement in assay performance was noted using the fluorogenic substrate Attophos compared with the chromogenic substrate p-nitrophenylphosphate. CONCLUSIONS: The detection system exhibiting the best characteristics with respect to cost, ease of use and overall performance in the detection of E. coli O157 in enrichment cultures from a variety of solid foods was based on the use of peroxidase as the indicator enzyme with the chromogenic substrate TMB. SIGNIFICANCE AND IMPACT OF THE STUDY: The polymyxin-ELISA provides a rapid, simple and inexpensive assay system for the detection of E. coli O157 in foods. PMID- 15548305 TI - Genetic diversity of Escherichia coli recovered from the oral cavity of beef cattle and their relatedness to faecal E. coli. AB - AIMS: To determine the genetic diversity of generic Escherichia coli recovered from the oral cavities of beef cattle and their relatedness to E. coli isolated from the faeces of cattle during pasture grazing and feedlot finishing. METHODS AND RESULTS: A total of 484 E. coli (248 oral and 236 faecal isolates) were obtained from eight beef cattle after 1 and 5 months of grazing on pasture and after 1 and 5 months in a feedlot. The random amplification of polymorphic DNA (RAPD) method was used to genetically characterize these isolates. The RAPD patterns showed that ca 60% of E. coli recovered from the oral cavities and faeces during pasture and feedlot shared a close genetic relatedness. A number of E. coli with unique RAPD types were also found either in the oral cavities or faeces. Most of the E. coli RAPD types recovered from the oral cavities were shared among animals, but there were also RAPD types which were unique to individual animals. The E. coli populations of the oral cavities were genetically diverse and changed over time. CONCLUSIONS: This study indicates that there are large numbers of E. coli carried in the oral cavities of beef cattle and those E. coli are closely related to strains found in the faeces. The oral cavities of cattle harbour a genetically diverse E. coli population. SIGNIFICANCE AND IMPACT OF THE STUDY: The oral cavity may be an important reservoir of enteric pathogens which may transfer to meat during carcass dressing. A better understanding of the molecular ecology of E. coli in cattle would assist the design of approaches to control pathogenic strains during beef production and processing. PMID- 15548306 TI - Differential inactivation of Listeria monocytogenes by D- and L-lactic acid. AB - AIMS: To determine inactivation of Listeria monocytogenes by the two lactic acid isomers. METHODS AND RESULTS: The survival of four strains with varying sensitivity to acid was determined following treatment with L- or D-lactic acid at 100 mmol l(-1) (pH 3.7) or HCl at pH 3.37. There was some, but not complete, similarity in the relative sensitivity of the four strains to the two types of acid. All strains were most sensitive to D-lactic acid, which gave 0.6-2.2 log units greater reduction than L-lactic acid midway in the inactivation curves. Even very low concentrations of the two isomers had an immediate effect on pH(i) which was identical for the two isomers. CONCLUSIONS: The results show that L. monocytogenes is more sensitive to D- than to L-lactic acid; however, this difference is less than the strain variation in L-lactic acid sensitivity. SIGNIFICANCE AND IMPACT OF THE STUDY: This work has implications for the application of lactic acid for food preservation as well as for the understanding of the antibacterial mechanisms of weak organic acids. PMID- 15548307 TI - Production of L-2,3-butanediol by a new pathway constructed in Escherichia coli. AB - AIMS: A metabolic pathway for L-2,3-butanediol (BD) as the main product has not yet been found. To rectify this situation, we attempted to produce L-BD from diacetyl (DA) by producing simultaneous expression of diacetyl reductase (DAR) and L-2,3-butanediol dehydrogenase (BDH) using transgenic bacteria, Escherichia coli JM109/pBUD-comb. METHODS AND RESULTS: The meso-BDH of Klebsiella pneumoniae was used for its DAR activity to convert DA to L-acetoin (AC) and the L-BDH of Brevibacterium saccharolyticum was used to reduce L-AC to L-BD. The respective gene coding each enzyme was connected in tandem to the MCS of pFLAG-CTC (pBUD comb). The divided addition of DA as a source, addition of 2% glucose, and the combination of static and shaking culture was effective for the production. CONCLUSIONS: L-BD (2200 mg l(-1)) was generated from 3000 mg l(-1) added of DA, which corresponded to a 73% conversion rate. Meso-BD as a by-product was mixed by 2% at most. SIGNIFICANCE AND IMPACT OF THE STUDY: An enzyme system for converting DA to L-BD was constructed with a view to using DA-producing bacteria in the future. PMID- 15548308 TI - The research imperative: fighting AIDS, TB and malaria. PMID- 15548309 TI - Citation patterns in tropical medicine journals. AB - Selections of most important journals in the field of tropical medicine have previously been identified with the help of resources such as bibliographical and citation databases. This article uses ISI's Journal Citation Reports (JSR) for 2002 to analyse the citation characteristics of the Tropical Medicine category. According to these data, this small but diverse group of 12 journals bestows some 40% more citations than it receives. Its six typical core journals tend to cite one another heavily, but they also refer a lot to multidisciplinary science and general medicine journals, and to infectious diseases and parasitology journals. Looking at the sources from which JCR's tropical medicine journals derive their citations, it is clear that in this reverse direction, the specialty's literature is still more concentrated. Apart from the typical core, this JCR category also contains a number of journals with more idiosyncratic citing patterns, focused on specialties such as paediatrics, a single disease (leprosy) and a representative of Latin American and Francophone biomedical science each. Implications of concentrated citedness and language biases are discussed briefly. This paper features a selection of bibliometric parameters relating to the tropical medicine journals and lists of the 80 journals most citing and cited by them. PMID- 15548310 TI - Fifty-year mortality trends in three rural African villages. AB - OBJECTIVE: To determine differential improvements in mortality rates according to age, and to discuss differences in neonatal and post-neonatal mortality rates, using longitudinal data from the past 50 years in three rural Gambian villages. METHODS: All villagers whose date of birth was recorded were followed up until death or October 1997. A Lexis expansion was performed to categorize age: early neonatal, late neonatal, infant, 1-4 years, 5-14 years and 15 or more years. Calendar time was divided into three periods -- prior to 1975, 1975-1984 and after 1985, representing different levels of clinical care. Cox regression and likelihood-ratio test were used to model the hazards ratios. RESULTS: There were 3981 subjects included in the analysis with a total of 59 002 person-years follow up. There was a dramatic decrease in neonatal (44 to 15), infant (162 to 36) and under-five mortality rates (397-66 per 1000) from pre-1975 to the present. The disproportionate decrease in the mortality rates means that neonatal deaths accounted for a greater percentage of post-natal deaths in later years. There was a marked seasonality in mortality rates with significantly more deaths in the 'hungry' season prior to 1975 (odds = 1.87; 95% CI = 1.62-2.17) and from 1975 to 1984 (odds = 1.84; 95% CI = 1.34-2.53). This seasonality of death has diminished in recent years (odds = 1.23; 95% CI = 0.85-1.76). CONCLUSION: Our analysis indicates a dramatic reduction in mortality rates and an attenuation in the seasonality of death. It is likely that the efficient implementation of basic currently available health measures was one of the key elements in achieving such a major reduction in mortality. PMID- 15548311 TI - Nationwide HIV prevalence survey in general population in Niger. AB - A national population-based survey was carried out in Niger in 2002 to assess HIV prevalence in the population aged 15-49 years. A two-stage cluster sampling was used and the blood specimens were collected on filter paper and tested according to an algorithm involving up to three diagnostic tests whenever appropriate. Testing was unlinked and anonymous. The refusal rate was 1.1% and 6056 blood samples were available for analysis. The adjusted prevalence of HIV was 0.87% (95% CI, 0.5-1.3%) and the 95% CI of the estimated number of infected individuals was 22 864-59 640. HIV-1 and HIV-2 represented, respectively, 95.6% and 2.9% of infections while dual infections represented 1.5%. HIV positivity rate was 1.0% in women and 0.7% in men. It was significantly higher among urban populations than among rural ones (respectively, 2.1% and 0.6%, P < 10(-6)). Using logistic regression, the variables significantly related to the risk of being tested positive for HIV were urban housing, increasing age and being either widowed or divorced. The estimate from the national survey was lower than the prevalence assessed from antenatal clinic data (2.8% in 2001). In the future, the representativeness of sentinel sites should be improved by increasing the representation of rural areas accounting for more than 80% of the population. Compared with other sub-Saharan countries, the HIV prevalence in Niger is still moderate. This situation represents a strong argument for enhancing prevention programmes and makes realistic the projects promoting an access to potent antiretroviral therapies for the majority. PMID- 15548312 TI - Melioidosis epidemiology and risk factors from a prospective whole-population study in northern Australia. AB - OBJECTIVES: The aims of this study were to describe the epidemiology of melioidosis in tropical northern Australia and to assess the importance of defined risk factors. METHODS: The data were taken from a 14-year prospective study of 364 cases of melioidosis in the 'Top End' of the Northern Territory. A whole-population logistic regression model was used to estimate the crude and adjusted relative risk (RR) for the defined risk factors. RESULTS: The mean age of the study population was 46.8 years, 264 (72.5%) were male, 178 (49%) were aboriginal Australians and 59 (16.2%) died from melioidosis. Average annual incidence was 19.6 cases per 100 000 population, with an estimated rate of 260 cases per 100,000 diabetics per year. Using a whole-population logistic regression model, the estimated crude and adjusted RR [95% confidence intervals (CI)] for melioidosis were 6.3 (5.1-7.8) and 4.0 (3.2-5.1) for those aged > or = 45 years, 2.3 (1.8-2.9) and 2.4 (1.9-3.0) for males, 2.9 (2.3-3.5) and 3.0 (2.3 4.0) for aboriginal Australians, 21.2 (17.1-26.3) and 13.1 (9.4-18.1) for diabetics, 2.7 (2.2-3.4) and 2.1 (1.6-2.6) for those with excess alcohol consumption, 6.8 (5.4-8.6) and 4.3 (3.4-5.5) for chronic lung disease and 6.7 (4.7-9.6) and 3.2 (2.2-4.8) for chronic renal disease, respectively. CONCLUSIONS: Diabetes, excess alcohol intake, chronic renal disease and chronic lung disease are each independent risk factors for melioidosis. In tropical northern Australia, male sex, aboriginal ethnicity and age of > or = 45 years are also independent predictors for melioidosis. Impaired polymorph function may be critical in the predisposition to melioidosis. PMID- 15548313 TI - Therapeutic efficacy of artemether-lumefantrine and artesunate-mefloquine for treatment of uncomplicated Plasmodium falciparum malaria in Luang Namtha Province, Lao People's Democratic Republic. AB - The efficacy of the six-dose regimen of artemether-lumefantrine was compared with the combination of artesunate and mefloquine in a randomised, comparative trial in Luang Namtha Province, Northern Laos. Of 1033 screened patients, 201 were positive for Plasmodium falciparum; 108 patients of all age groups (2-66 years) with acute, uncomplicated P. falciparum malaria were enrolled in the study, 100 of whom were followed-up for 42 days. Fifty-three patients received artemether lumefantrine and 55 received artesunante-mefloquine. Both drug combinations induced rapid clearance of parasites and malaria symptoms; there was no significant difference in the initial therapeutic response parameters. Both regimes were well tolerated. After 42 days, cure rates were 93.6% (95% CI = 82.5 98.7%; 44 of 47 patients) for artemether-lumefantrine and 100% (95% CI = 93.3 100.0%; 53 of 53 patients) for artesunate-mefloquine. The results show the excellent efficacy and tolerability of both artemether-lumefantrine and artesunate-mefloquine in Northern Laos. PMID- 15548314 TI - Comparison of chloroquine, sulfadoxine/pyrimethamine, mefloquine and mefloquine artesunate for the treatment of falciparum malaria in Kachin State, North Myanmar. AB - Multi-drug resistant falciparum malaria is widespread in Asia. In Thailand, Cambodia and Vietnam the national protocols have changed largely to artesunate combined treatment regimens but elsewhere in East and South Asia chloroquine (CQ) and sulfadoxine-pyrimethamine (SP) are still widely recommended by national malaria control programmes. In Kachin State, northern Myanmar, an area of low seasonal malaria transmission, the efficacy of CQ (25 mg base/kg) and SP (1.25/25 mg/kg), the nationally recommended treatments at the time, were compared with mefloquine alone (M; 15 mg base/kg) and mefloquine combined with artesunate (MA; 15:4 mg/kg). An open randomized controlled trial enrolled 316 patients with uncomplicated Plasmodium falciparum malaria, stratified prospectively into three age-groups. Early treatment failures (ETF) occurred in 41% (32/78) of CQ treated patients and in 24% of patients treated with SP (18/75). In young children the ETF rates were 87% after CQ and 35% after SP. Four children (two CQ, two SP) developed symptoms of cerebral malaria within 3 days after treatment. By day 42, failure rates (uncorrected for reinfections) had increased to 79% for CQ and 81% for SP. ETF rates were 2.5% after treatment with M and 3.9% after treatment with MA (P > 0.2). Overall uncorrected treatment failure rates at day 42 following M and MA were 23% and 21%, respectively. Chloroquine and SP are completely ineffective for the treatment of falciparum malaria in northern Myanmar. Mefloquine treatment is much more effective, but three day combination regimens with artesunate will be needed for optimum efficacy and protection against resistance. PMID- 15548315 TI - Local fever illness classifications: implications for home management of malaria strategies. AB - BACKGROUND: The Ugandan Ministry of Health has adopted the WHO Home Based Fever Management strategy (HBM) to improve access to antimalarial drugs for prompt (<24 h) presumptive treatment of all fevers in children under 5 years. Village volunteers will distribute pre-packed antimalarials free of charge to caretakers of febrile children 2 months to 5 years ('Homapaks'). OBJECTIVE: To explore the local understanding and treatment practices for childhood fever illnesses and discuss implications for the HBM strategy. METHODS: Focus Group Discussions were held with child caretakers in three rural communities in Kasese district, West Uganda, and analysed for content in respect to local illness classifications and associated treatments for childhood fevers. RESULTS: Local understanding of fever illnesses and associated treatments was complex. Some fever illness classifications were more commonly mentioned, including 'Fever of Mosquito', 'Chest Problem', 'the Disease', 'Stomach Wounds' and 'Jerks', all of which could be biomedical malaria. Although caretakers refer to all these classifications as 'fever' treatment differed; some were seen as requiring urgent professional western treatment and others were considered severe but 'non-western' and would preferentially be treated with traditional remedies. CONCLUSIONS: The HBM strategy does not address local community understanding of 'fever' and its influence on treatment. While HBM improves drug access, Homapaks are likely to be used for only those fevers where 'western' treatment is perceived appropriate, implying continued delayed and under-treatment of potential malaria. Hence, HBM strategies also need to address local perceptions of febrile illness and adapt information and training material accordingly. PMID- 15548316 TI - Observations on false positive reactions in the rapid NOW Filariasis card test. AB - The NOW Filariasis card test is a useful tool for rapid field diagnosis of Wuchereria bancrofti infection, based on detection of specific circulating filarial antigen (CFA) in the patients' blood. Concern has been raised that a high proportion of infection negative individuals develop false positive reactions in these tests when the test cards are left for a prolonged period before being examined. We carried out a survey in an endemic Tanzanian village to investigate this phenomenon. Individuals who were positive in the NOW Filariasis test at 10 min after specimen application were also positive in the TropBio ELISA for CFA, and thus appeared to be truly positive. Many of the test cards that were negative at 10 min developed a positive line later, but these lines appeared to be falsely positive when the TropBio test was used as the gold standard. Close examination revealed that true and false positivity lines could be distinguished on their shape and colour. The study thus reaffirmed that test cards should be read after 10 min to avoid false positives, but it also indicated that experienced test card users should be able to make a correct diagnosis even at a later time. PMID- 15548317 TI - A 22-mer primer enhances discriminatory power of AP-PCR fingerprinting technique in characterization of leptospires. AB - OBJECTIVES: To evaluate the discriminatory power and usefulness of arbitrarily primed-polymerase chain reaction (AP-PCR) characterization of leptospires with M16 primer. METHODS: AP-PCR fingerprints of 20 reference strains of Leptospira representing 20 different serovars belonging to seven genospecies (Leptospira interrogans, 11; L. noguchii, 2; L. borgpetersenii, 1; L. santarosai, 2; L. biflexa, 2; L. kirschneri, 1; L. weilii, 1) were generated by employing M16 primer. Fingerprints generated with this primer were compared with those generated with two other commonly used primers PB1, and L10. An attempt was also made to type 20 leptospiral isolates with the M16 primer. RESULTS AND CONCLUSION: Fingerprints with M16 primer could not only differentiate between strains of different genospecies, but also between strains of the same genospecies belonging to different serovars. While two commonly used primers (PB1 and L10) failed to discriminate between some of the different serovars belonging to the same genospecies, this primer was able to generate discriminatory fingerprints for all strains tested. All 20 Leptospira isolates, recovered from patients in Andaman Islands, could also be typed by fingerprints generated with the M16 primer. The discriminatory power of M16 primer adds more specificity to the rapidity of this system of characterization and can be used as an excellent tool in epidemiological studies on Leptospira. PMID- 15548318 TI - Is there an association between bacteriological drinking water quality and childhood diarrhoea in developing countries? AB - To investigate the association between bacteriological drinking water quality and incidence of diarrhoea, we conducted a 1-year prospective study in the southern Punjab, Pakistan. Diarrhoea episodes, drinking water sources and drinking water quality were monitored weekly among children younger than 5 years in 200 households. We found no association between the incidence of childhood diarrhoea and the number of Escherichia coli in the drinking water sources (the public domain). A possible trend was seen relating the number of E. coli in the household storage containers (the domestic domain) and diarrhoea incidence, but this did not reach statistical significance. Faecal contamination levels in household water containers were generally high even when the source water was of good quality. Under conditions such as this, it is questionable whether public water treatment will have a significant impact on the incidence of endemic childhood diarrhoea. PMID- 15548319 TI - Treatment of maternal syphilis in rural South Africa: effect of multiple doses of benzathine penicillin on pregnancy loss. AB - OBJECTIVES: Despite few data, the treatment of syphilis in pregnant women using a single dose of benzathine penicillin is the standard of care in many resource poor settings. We examined the effect of various doses of benzathine penicillin on pregnancy loss among women with a positive Rapid Plasma Reagin (RPR) test result in a rural South African district. METHODS: All pregnant women making their first antenatal care visit during pregnancy were screened for syphilis using the RPR test. Those testing positive were counselled to receive three weekly doses of benzathine penicillin, and received a partner notification card. Pregnancy outcomes were determined from facility records or home visits where necessary. RESULTS: Of 8917 women screened, 1043 (12%) had reactive syphilis serology; of those with titre data available, 30% had titres of 1:8 or greater. While 41% (n = 430) of women received all three doses as counselled, 30% (n = 312) received only one dose, and 20% (n = 207) did not return to the clinic to receive treatment. Among the 947 women with pregnancy outcome data available, there were 17 miscarriages and 48 perinatal deaths observed. There was a strong trend towards reduced risk of pregnancy loss among women receiving multiple doses of penicillin (adjusted OR for perinatal mortality for each additional dose received, 0.63; 95% CI, 0.48-0.84). CONCLUSIONS: While this association requires further investigation, these results suggest that there may be substantial benefit to providing multiple doses of benzathine penicillin to treat maternal syphilis in this setting. PMID- 15548320 TI - [Profile of members and non members of mutual health insurance system in Rwanda: the case of the health district of Kabutare]. AB - The establishment of mutual health insurance systems is one of the priorities of the Rwandan government. Pilot studies have been conducted in three districts of the country. Nonetheless, after 4 years of implementation (1999-2003), the population coverage by these insurance systems remains relatively low. A cross sectional study of 1042 households in the Kabutare health district allowed for a comparison of socio-economic and demographic variables, and the medical, surgical, gynaecological, and obstetrical history of health insurance scheme members and non-members. The results of the study demonstrate that the distribution of members and non-members is similar in terms of sex, marital status, professional status and medical history. However, larger households (more than five members) and those having a relatively higher income (more than USD 230 per annum) are more likely to be insured than other households. Members of the mutual health insurance use more the health services than non-members, spend less on health care and increasingly maintain membership. The study emphasizes the relevance to further promote mutual health insurance, but also points to the need for mechanisms to ensure financial access for the poor rural population. PMID- 15548321 TI - Cultural concepts of tuberculosis and gender among the general population without tuberculosis in rural Maharashtra, India. AB - Gender-specific patterns of experience, meaning, and behaviour for tuberculosis (TB) require consideration to guide control programmes. To clarify concepts of gender, culture, and TB in a rural endemic population of Maharashtra, India, this study of 80 men and 80 women employed qualitative and quantitative methods of cultural epidemiology, using a locally adapted semi-structured Explanatory Model Interview Catalogue (EMIC) interviews are instruments for cultural epidemiological study of the distribution of illness-related experiences, meanings, and behaviours. This interview queried respondents without active disease about vignettes depicting a man and woman with typical features of TB. Emotional and social symptoms were frequently reported for both vignettes, but more often considered most distressing for the female vignette; specified problems included arranging marriages, social isolation, and inability to care for children and family. Job loss and reduced income were regarded most troubling for the male vignette. Men and women typically identified sexual experience as the cause of TB for opposite-sex vignettes. With wider access to information about TB, male respondents more frequently recommended allopathic doctors and specialty services. Discussion considers the practical significance of gender specific cultural concepts of TB. PMID- 15548322 TI - Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium. AB - BACKGROUND: Old, stationary cultures of Mycobacterium tuberculosis contain a majority of bacteria that can grow in broth cultures but cannot grow on solid medium plates. These may be in a non-replicating, dormant growth phase. We hypothesised that a similar population might be present in chronic, murine tuberculosis. METHODS: Estimates of the numbers of viable M. tuberculosis, strain H37Rv, in the spleens and lungs of mice in a 7-day acute infection and in a 10 month chronic infection were made by conventional plate counts and, as broth counts, by noting presence or absence of growth in serial replicate dilutions in liquid medium. RESULTS: Plate and broth counts in 6 mice gave similar mean values in the acute infection, 7 days after infection. However, the broth counts were much higher in 36 mice with a chronic infection at 10 months. Broth counts averaged 5.290 log10 cfu /organ from spleens and 5.523 log10 cfu/organ from lungs, while plate counts were 3.858 log10 cfu/organ from spleens and 3.662 log10 cfu/organ from lungs, indicating that the total bacterial population contained only 3.7% bacilli in spleens and 1.4% bacilli in lungs, capable of growth on plates. CONCLUSION: The proportion growing on plates might be a measure of the "dormancy" of the bacilli equally applicable to cultural and animal models. PMID- 15548323 TI - Threshold intensity factors as lower boundaries for crack propagation in ceramics. AB - BACKGROUND: Slow crack growth can be described in a v (crack velocity) versus KI (stress intensity factor) diagram. Slow crack growth in ceramics is attributed to corrosion assisted stress at the crack tip or at any pre-existing defect in the ceramic. The combined effect of high stresses at the crack tip and the presence of water or body fluid molecules (reducing surface energy at the crack tip) induces crack propagation, which eventually may result in fatigue. The presence of a threshold in the stress intensity factor, below which no crack propagation occurs, has been the subject of important research in the last years. The higher this threshold, the higher the reliability of the ceramic, and consequently the longer its lifetime. METHODS: We utilize the Irwin K-field displacement relation to deduce crack tip stress intensity factors from the near crack tip profile. Cracks are initiated by indentation impressions. The threshold stress intensity factor is determined as the time limit of the tip stress intensity when the residual stresses have (nearly) disappeared. RESULTS: We determined the threshold stress intensity factors for most of the all ceramic materials presently important for dental restorations in Europe. Of special significance is the finding that alumina ceramic has a threshold limit nearly identical with that of zirconia. CONCLUSION: The intention of the present paper is to stress the point that the threshold stress intensity factor represents a more intrinsic property for a given ceramic material than the widely used toughness (bend strength or fracture toughness), which refers only to fast crack growth. Considering two ceramics with identical threshold limits, although with different critical stress intensity limits, means that both ceramics have identical starting points for slow crack growth. Fast catastrophic crack growth leading to spontaneous fatigue, however, is different. This growth starts later in those ceramic materials that have larger critical stress intensity factors. PMID- 15548324 TI - Transport lattice models of heat transport in skin with spatially heterogeneous, temperature-dependent perfusion. AB - BACKGROUND: Investigation of bioheat transfer problems requires the evaluation of temporal and spatial distributions of temperature. This class of problems has been traditionally addressed using the Pennes bioheat equation. Transport of heat by conduction, and by temperature-dependent, spatially heterogeneous blood perfusion is modeled here using a transport lattice approach. METHODS: We represent heat transport processes by using a lattice that represents the Pennes bioheat equation in perfused tissues, and diffusion in nonperfused regions. The three layer skin model has a nonperfused viable epidermis, and deeper regions of dermis and subcutaneous tissue with perfusion that is constant or temperature dependent. Two cases are considered: (1) surface contact heating and (2) spatially distributed heating. The model is relevant to the prediction of the transient and steady state temperature rise for different methods of power deposition within the skin. Accumulated thermal damage is estimated by using an Arrhenius type rate equation at locations where viable tissue temperature exceeds 42 degrees C. Prediction of spatial temperature distributions is also illustrated with a two-dimensional model of skin created from a histological image. RESULTS: The transport lattice approach was validated by comparison with an analytical solution for a slab with homogeneous thermal properties and spatially distributed uniform sink held at constant temperatures at the ends. For typical transcutaneous blood gas sensing conditions the estimated damage is small, even with prolonged skin contact to a 45 degrees C surface. Spatial heterogeneity in skin thermal properties leads to a non-uniform temperature distribution during a 10 GHz electromagnetic field exposure. A realistic two-dimensional model of the skin shows that tissue heterogeneity does not lead to a significant local temperature increase when heated by a hot wire tip. CONCLUSIONS: The heat transport system model of the skin was solved by exploiting the mathematical analogy between local thermal models and local electrical (charge transport) models, thereby allowing robust, circuit simulation software to obtain solutions to Kirchhoff's laws for the system model. Transport lattices allow systematic introduction of realistic geometry and spatially heterogeneous heat transport mechanisms. Local representations for both simple, passive functions and more complex local models can be easily and intuitively included into the system model of a tissue. PMID- 15548325 TI - CP-31398, a putative p53-stabilizing molecule tested in mammalian cells and in yeast for its effects on p53 transcriptional activity. AB - BACKGROUND: CP-31398 is a small molecule that has been reported to stabilize the DNA-binding core domain of the human tumor suppressor protein p53 in vitro. The compound was also reported to function as a potential anti-cancer drug by rescuing the DNA-binding activity and, consequently, the transcription activation function of mutant p53 protein in mammalian tissue culture cells and in mice. RESULTS: We performed a series of gene expression experiments to test the activity of CP-31398 in yeast and in human cell cultures. With these cell-based assays, we were unable to detect any specific stimulation of mutant p53 activity by this compound. Concentrations of CP-31398 that were reported to be active in the published work were highly toxic to the human H1299 lung carcinoma and Saos-2 cell lines in our experiments. CONCLUSION: In our experiments, the small molecule CP-31398 was unable to reactivate mutant p53 protein. The results of our in vivo experiments are in agreement with the recently published biochemical analysis of CP-31398 showing that this molecule does not bind p53 as previously claimed, but intercalates into DNA. PMID- 15548326 TI - Hand-carried ultrasound performed at bedside in cardiology inpatient setting - a comparative study with comprehensive echocardiography. AB - BACKGROUND: Hand-carried ultrasound (HCU) devices have been demonstrated to improve the diagnosis of cardiac diseases over physical examination, and have the potential to broaden the versatility in ultrasound application. The role of these devices in the assessment of hospitalized patients is not completely established. In this study we sought to perform a direct comparison between bedside evaluation using HCU and comprehensive echocardiography (CE), in cardiology inpatient setting. METHODS: We studied 44 consecutive patients (mean age 54 +/- 18 years, 25 men) who underwent bedside echocardiography using HCU and CE. HCU was performed by a cardiologist with level-2 training in the performance and interpretation of echocardiography, using two-dimensional imaging, color Doppler, and simple calliper measurements. CE was performed by an experienced echocardiographer (level-3 training) and considered as the gold standard. RESULTS: There were no significant differences in cardiac chamber dimensions and left ventricular ejection fraction determined by the two techniques. The agreement between HCU and CE for the detection of segmental wall motion abnormalities was 83% (Kappa = 0.58). There was good agreement for detecting significant mitral valve regurgitation (Kappa = 0.85), aortic regurgitation (kappa = 0.89), and tricuspid regurgitation (Kappa = 0.74). A complete evaluation of patients with stenotic and prosthetic dysfunctional valves, as well as pulmonary hypertension, was not possible using HCU due to its technical limitations in determining hemodynamic parameters. CONCLUSION: Bedside evaluation using HCU is helpful for assessing cardiac chamber dimensions, left ventricular global and segmental function, and significant valvular regurgitation. However, it has limitations regarding hemodynamic assessment, an important issue in the cardiology inpatient setting. PMID- 15548327 TI - The "Transport Specificity Ratio": a structure-function tool to search the protein fold for loci that control transition state stability in membrane transport catalysis. AB - BACKGROUND: In establishing structure-function relationships for membrane transport proteins, the interpretation of phenotypic changes can be problematic, owing to uncertainties in protein expression levels, sub-cellular localization, and protein-folding fidelity. A dual-label competitive transport assay called "Transport Specificity Ratio" (TSR) analysis has been developed that is simple to perform, and circumvents the "expression problem," providing a reliable TSR phenotype (a constant) for comparison to other transporters. RESULTS: Using the Escherichia coli GABA (4-aminobutyrate) permease (GabP) as a model carrier, it is demonstrated that the TSR phenotype is largely independent of assay conditions, exhibiting: (i) indifference to the particular substrate concentrations used, (ii) indifference to extreme changes (40-fold) in transporter expression level, and within broad limits (iii) indifference to assay duration. The theoretical underpinnings of TSR analysis predict all of the above observations, supporting that TSR has (i) applicability in the analysis of membrane transport, and (ii) particular utility in the face of incomplete information on protein expression levels and initial reaction rate intervals (e.g., in high-throughput screening situations). The TSR was used to identify gab permease (GabP) variants that exhibit relative changes in catalytic specificity (kcat/Km) for [14C]GABA (4 aminobutyrate) versus [3H]NA (nipecotic acid). CONCLUSIONS: The TSR phenotype is an easily measured constant that reflects innate molecular properties of the transition state, and provides a reliable index of the difference in catalytic specificity that a carrier exhibits toward a particular pair of substrates. A change in the TSR phenotype, called a Delta(TSR), represents a specificity shift attributable to underlying changes in the intrinsic substrate binding energy (DeltaGb) that translocation catalysts rely upon to decrease activation energy (Delta G(T)(++). TSR analysis is therefore a structure-function tool that enables parsimonious scanning for positions in the protein fold that couple to the transition state, creating stability and thereby serving as functional determinants of catalytic power (efficiency, or specificity). PMID- 15548328 TI - The effect of short-duration sub-maximal cycling on balance in single-limb stance in patients with anterior cruciate ligament injury: a cross-sectional study. AB - BACKGROUND: It has previously been shown that an anterior cruciate ligament (ACL) injury may lead to impaired postural control, and that the ability to maintain postural control is decreased by fatigue in healthy subjects. To our knowledge, no studies have reported the effect of fatigue on postural control in subjects with ACL injury. This study was aimed at examining the effect of fatigue on balance in single-limb stance in subjects with ACL injury, and to compare the effects, and the ability to maintain balance, with that of a control group of uninjured subjects. METHODS: Thirty-six patients with unilateral, non-operated, non-acute ACL injury, and 24 uninjured subjects were examined with stabilometry before (pre-exercise) and immediately after (post-exercise) short-duration, sub maximal cycling. In addition, the post-exercise measurements were compared, to evaluate the instantaneous ability to maintain balance and any possible recovery. The amplitude and average speed of center of pressure movements were registered in the frontal and sagittal planes. The paired t-test was used for the intra group comparisons, and the independent t-test for the inter-group comparisons, with Bonferroni correction for multiple comparisons. RESULTS: No differences were found in the effects of exercise between the patients and the controls. Analysis of the post-exercise measurements revealed greater effects or a tendency towards greater effects on the injured leg than in the control group. The average speed was lower among the patients than in the control group. CONCLUSIONS: The results of the present study showed no differences in the effects of exercise between the patients and the controls. However, the patients seemed to react differently regarding ability to maintain balance in single-limb stance directly after exercise than the control group. The lower average speed among the patients may be an expression of different neuromuscular adaptive strategies than in uninjured subjects. PMID- 15548329 TI - Elevated responses to constant facial emotions in different faces in the human amygdala: an fMRI study of facial identity and expression. AB - BACKGROUND: Human faces provide important signals in social interactions by inferring two main types of information, individual identity and emotional expression. The ability to readily assess both, the variability and consistency among emotional expressions in different individuals, is central to one's own interpretation of the imminent environment. A factorial design was used to systematically test the interaction of either constant or variable emotional expressions with constant or variable facial identities in areas involved in face processing using functional magnetic resonance imaging. RESULTS: Previous studies suggest a predominant role of the amygdala in the assessment of emotional variability. Here we extend this view by showing that this structure activated to faces with changing identities that display constant emotional expressions. Within this condition, amygdala activation was dependent on the type and intensity of displayed emotion, with significant responses to fearful expressions and, to a lesser extent so to neutral and happy expressions. In contrast, the lateral fusiform gyrus showed a binary pattern of increased activation to changing stimulus features while it was also differentially responsive to the intensity of displayed emotion when processing different facial identities. CONCLUSIONS: These results suggest that the amygdala might serve to detect constant facial emotions in different individuals, complementing its established role for detecting emotional variability. PMID- 15548330 TI - Saposin C promotes survival and prevents apoptosis via PI3K/Akt-dependent pathway in prostate cancer cells. AB - BACKGROUND: In addition to androgens, growth factors are also implicated in the development and neoplastic growth of the prostate gland. Prosaposin is a potent neurotrophic molecule. Homozygous inactivation of prosaposin in mice has led to the development of a number of abnormalities in the male reproductive system, including atrophy of the prostate gland and inactivation of mitogen-activated protein kinase (MAPK) and Akt in prostate epithelial cells. We have recently reported that prosaposin is expressed at a higher level by androgen-independent (AI) prostate cancer cells as compared to androgen-sensitive prostate cancer cells or normal prostate epithelial and stromal cells. In addition, we have demonstrated that a synthetic peptide (prosaptide TX14A), derived from the trophic sequence of the saposin C domain of prosaposin, stimulated cell proliferation, migration and invasion and activated the MAPK signaling pathway in prostate cancer cells. The biological significances of saposin C and prosaposin in prostate cancer are not known. RESULTS: Here, we report that saposin C, in a cell type-specific and dose-dependent manner, acts as a survival factor, activates the Akt-signaling pathway, down-modulates caspase-3, -7, and -9 expression and/or activity, and decreases the cleaved nuclear substrate of caspase-3 in prostate cancer cells under serum-starvation stress. In addition, prosaptide TX14A, saposin C, or prosaposin decreased the growth-inhibitory effect, caspase-3/7 activity, and apoptotic cell death induced by etoposide. We also discovered that saposin C activates the p42/44 MAP kinase pathway in a pertussis toxin-sensitive and phosphatidylinositol 3-kinase (PI3K) /Akt-dependent manner in prostate cancer cells. Our data also show that the anti-apoptotic activity of saposin C is at least partially mediated via PI3K/Akt signaling pathway. CONCLUSION: We postulate that as a mitogenic, survival, and anti apoptotic factor for prostate cancer cells, saposin C or prosaposin may contribute to prostate carcinogenesis at its early androgen-dependent or metastatic AI state. PMID- 15548331 TI - "Tied together like a woven hat:" Protective pathways to Alaska native sobriety. AB - BACKGROUND: The People Awakening Project (1RO1 AA 11446-03) had two purposes, completed in Phase I and Phase II of the project. The purpose of Phase I was to complete a qualitative study; the research objective was discovery oriented with the specific aim of identification of protective and recovery factors in Alaska Native sobriety. Results were used to develop a heuristic model of protective and recovery factors, and measures based on these factors. The research objective of Phase II was to pilot these measures and provide initial validity data. METHODS: Phase I utilized a life history methodology. People Awakening interviewed a convenience sample of 101 Alaska Natives who had either recovered from alcoholism (n = 58) or never had a drinking problem (n = 43). This later group included both lifetime abstainers (LAs) and non-problem drinkers (NPs). Life histories were transcribed and analyzed using grounded theory and consensual data analytic procedures within a participatory action research framework. Analyses were utilized to generate heuristic models of protection and recovery from alcohol abuse among Alaska Natives. RESULTS: Analyses generated a heuristic model of protective factors from alcohol abuse. The resulting multilevel and multi factorial model describes interactive and reciprocal influences of (a) individual, family, and community characteristics; (b) trauma and the individual and contextual response to trauma, (c) experimental substance use and the person's social environment; and (d) reflective processes associated with a turning point, or a life decision regarding sobriety. The importance of cultural factors mediating all these protective processes is emphasized. For NPs, the resilience process drew from personal stores of self-confidence, self-efficacy, and self-mastery that derived from ability to successfully maneuver within stressful or potentially traumatizing environments. In contrast, for many LAs, efficacy was instead described in more socially embedded terms better understood as communal mastery. One style of mastery is more associated with individualistic orientations, the other with more collectivistic. Future research is needed regarding the generalizeability of this group difference. CONCLUSIONS: Results suggest that preventative interventions should focus on intervening simultaneously at the community, family, and individual levels to build resilience and protective factors at each level. Of particular importance is the building of reflexivity along with other cognitive processes that allow the individual to think through problems and to reach a life decision to not abuse alcohol. PMID- 15548332 TI - Historical measures of social context in life course studies: retrospective linkage of addresses to decennial censuses. AB - BACKGROUND: There is evidence of a contribution of early life socioeconomic exposures to the risk of chronic diseases in adulthood. However, extant studies investigating the impact of the neighborhood social environment on health tend to characterize only the current social environment. This in part may be due to complexities involved in obtaining and geocoding historical addresses. The Life Course Socioeconomic Status, Social Context, and Cardiovascular Disease Study collected information on childhood (1930-1950) and early adulthood (1960-1980) place of residence from 12,681 black and white middle-aged and older men and women from four U.S. communities to link participants with census-based socioeconomic indicators over the life course. RESULTS: Most (99%) participants were linked to 1930-50 county level socioeconomic census data (the smallest level of aggregation universally available during this time period) corresponding to childhood place of residence. Linkage did not vary by race, gender, birth cohort, or level of educational attainment. A commercial geocoding vendor processed participants' self-reported street addresses for ages 30, 40, and 50. For 1970 and 1980 censuses, spatial coordinates were overlaid onto shape files containing census tract boundaries; for 1960 no shape files existed and comparability files were used. Several methods were tested for accuracy and to increase linkage. Successful linkage to historical census tracts varied by census (66% for 1960, 76% for 1970, 85% for 1980). This compares to linkage rates of 94% for current addresses provided by participants over the course of the ARIC examinations. CONCLUSION: There are complexities and limitations in characterizing the past social context. However, our results suggest that it is feasible to characterize the earlier social environment with known levels of measurement error and that such an approach should be considered in future studies. PMID- 15548333 TI - Genome structure and transcriptional regulation of human coronavirus NL63. AB - BACKGROUND: Two human coronaviruses are known since the 1960s: HCoV-229E and HCoV OC43. SARS-CoV was discovered in the early spring of 2003, followed by the identification of HCoV-NL63, the fourth member of the coronaviridae family that infects humans. In this study, we describe the genome structure and the transcription strategy of HCoV-NL63 by experimental analysis of the viral subgenomic mRNAs. RESULTS: The genome of HCoV-NL63 has the following gene order: 1a-1b-S-ORF3-E-M-N. The GC content of the HCoV-NL63 genome is extremely low (34%) compared to other coronaviruses, and we therefore performed additional analysis of the nucleotide composition. Overall, the RNA genome is very low in C and high in U, and this is also reflected in the codon usage. Inspection of the nucleotide composition along the genome indicates that the C-count increases significantly in the last one-third of the genome at the expense of U and G. We document the production of subgenomic (sg) mRNAs coding for the S, ORF3, E, M and N proteins. We did not detect any additional sg mRNA. Furthermore, we sequenced the 5' end of all sg mRNAs, confirming the presence of an identical leader sequence in each sg mRNA. Northern blot analysis indicated that the expression level among the sg mRNAs differs significantly, with the sg mRNA encoding nucleocapsid (N) being the most abundant. CONCLUSIONS: The presented data give insight into the viral evolution and mutational patterns in coronaviral genome. Furthermore our data show that HCoV-NL63 employs the discontinuous replication strategy with generation of subgenomic mRNAs during the (-) strand synthesis. Because HCoV-NL63 has a low pathogenicity and is able to grow easily in cell culture, this virus can be a powerful tool to study SARS coronavirus pathogenesis. PMID- 15548334 TI - Permissive human cytomegalovirus infection of a first trimester extravillous cytotrophoblast cell line. AB - Human cytomegalovirus (HCMV) is the leading cause of congenital viral infection in the United States and Europe. Despite the significant morbidity associated with prenatal HCMV infection, little is known about how the virus infects the fetus during pregnancy. To date, primary human cytotrophoblasts (CTBs) have been utilized to study placental HCMV infection and replication; however, the minimal mitotic potential of these cells restricts experimentation to a few days, which may be problematic for mechanistic studies of the slow-replicating virus. The aim of this study was to determine whether the human first trimester CTB cell line SGHPL-4 was permissive for HCMV infection and therefore could overcome such limitations. HCMV immediate early (IE) protein expression was detected as early as 3 hours post-infection in SGHPL-4 cells and progressively increased as a function of time. HCMV growth assays revealed the presence of infectious virus in both cell lysates and culture supernatants, indicating that viral replication and the release of progeny virus occurred. Compared to human fibroblasts, viral replication was delayed in CTBs, consistent with previous studies reporting delayed viral kinetics in HCMV-infected primary CTBs. These results indicate that SGHPL-4 cells are fully permissive for the complete HCMV replicative cycle. Our findings suggest that these cells may serve as useful tools for future mechanistic studies of HCMV pathogenesis during early pregnancy. PMID- 15548336 TI - Out of the christmas box. PMID- 15548337 TI - Correlates of fruit and vegetable intake among Norwegian schoolchildren: parental and self-reports. AB - OBJECTIVES: To identify correlates of 6th and 7th graders' (age 10-12 years) fruit and vegetable intake, to investigate parent-child correlations of fruit and vegetable intake, and to compare parents' and children's reports of children's accessibility, skills and preferences with respect to fruit and vegetables. DESIGN: The results presented are based on the baseline survey of the 'Fruits and Vegetables Make the Marks Project', where 38 schools participated. SETTING: Fruit and vegetable intake was measured by food frequency questions. Theoretical factors, based on Social Cognitive Theory, potentially correlated to intake were measured, including behavioural skills, accessibility, modelling, intention, preferences, self-efficacy and awareness of 5-a-day recommendations. SUBJECTS: In total, 1950 (participation rate 85%) 6th and 7th graders and 1647 of their parents participated. RESULTS: Overall, 34% of the variance in the pupils' reported fruit and vegetable intake was explained by the measured factors. The strongest correlates to fruit and vegetable intake were preferences and accessibility. The correlation between the children's and their parents' fruit and vegetable intake was 0.23. The parents perceived their children's accessibility to be better than what was reported by the children (P<0.01), while the children reported their skills to be better than what was perceived by their parents (P<0.01). CONCLUSION: The results from this study clearly point to a need for nutrition interventions aimed at parents. An important next step will be to investigate whether the identified correlates predict future fruit and vegetable intake, and whether they mediate any changes in intake in an intervention study. PMID- 15548338 TI - Correlates of diet quality in the Quebec population. AB - OBJECTIVE: To determine the correlates of a high-fat diet in urban, suburban and rural areas of Quebec, Canada. DESIGN: A secondary analysis of data collected as part of a 5-year multi-factorial, multi-setting, community-intervention project. SETTING: Urban, suburban and rural settings of the province of Quebec, 1997. SUBJECTS: Data were analysed from a sample of 5214 participants (2227 males, 2987 females). A food-frequency questionnaire was completed and a global index of food quality was calculated. Logistic regression was used to identify correlates of a diet high in total fats, saturated fat and cholesterol. RESULTS: In both genders, lower level of education, smoking status, French and English languages compared with other languages spoken at home, and a rural environment were associated with poor diet quality. Having no intention to eat low-fat dairy products more often was associated with a high-fat diet. In men, obesity (body mass index >/=30 kg m( 2)) and absence of reported health problems were correlates of a high-fat diet, while, in women, lower physical activity was a correlate. CONCLUSIONS: Future health interventions in Quebec should target people with low education, smokers and those living in a rural environment. Obese men and sedentary women should have access to specific dietetic resources. PMID- 15548339 TI - Dietary patterns in middle-aged Irish men and women defined by cluster analysis. AB - OBJECTIVES: To identify and characterise dietary patterns in a middle-aged Irish population sample and study associations between these patterns, sociodemographic and anthropometric variables and major risk factors for cardiovascular disease. DESIGN: A cross-sectional study. SUBJECTS AND METHODS: A group of 1473 men and women were sampled from 17 general practice lists in the South of Ireland. A total of 1018 attended for screening, with a response rate of 69%. Participants completed a detailed health and lifestyle questionnaire and provided a fasting blood sample for glucose, lipids and homocysteine. Dietary intake was assessed using a standard food-frequency questionnaire adapted for use in the Irish population. The food-frequency questionnaire was a modification of that used in the UK arm of the European Prospective Investigation into Cancer study, which was based on that used in the US Nurses' Health Study. Dietary patterns were assessed primarily by K-means cluster analysis, following initial principal components analysis to identify the seeds. RESULTS: Three dietary patterns were identified. These clusters corresponded to a traditional Irish diet, a prudent diet and a diet characterised by high consumption of alcoholic drinks and convenience foods. Cluster 1 (Traditional Diet) had the highest intakes of saturated fat (SFA), monounsaturated fat (MUFA) and percentage of total energy from fat, and the lowest polyunsaturated fat (PUFA) intake and ratio of polyunsaturated to saturated fat (P:S). Cluster 2 (Prudent Diet) was characterised by significantly higher intakes of fibre, PUFA, P:S ratio and antioxidant vitamins (vitamins C and E), and lower intakes of total fat, MUFA, SFA and cholesterol. Cluster 3 (Alcohol & Convenience Foods) had the highest intakes of alcohol, protein, cholesterol, vitamin B(12), vitamin B(6), folate, iron, phosphorus, selenium and zinc, and the lowest intakes of PUFA, vitamin A and antioxidant vitamins (vitamins C and E). There were significant differences between clusters in gender distribution, smoking status, physical activity, body mass index, waist circumference and serum homocysteine concentrations. CONCLUSION: In this general population sample, cluster analysis methods yielded two major dietary patterns: prudent and traditional. The prudent dietary pattern is associated with other health-seeking behaviours. Study of dietary patterns will help elucidate links between diet and disease and contribute to the development of healthy eating guidelines for health promotion. PMID- 15548340 TI - Differences in food habits and cardiovascular disease risk factors among Native Americans with and without diabetes: the Inter-Tribal Heart Project. AB - OBJECTIVE: To examine differences in food habits among Native Americans with and without diabetes. DESIGN: A cross-sectional epidemiological study in which participants underwent a physical examination and answered an extensive interviewer-administered questionnaire to assess differences in food servings, preparation and eating habits. SETTING/PARTICIPANTS: Participants aged >/=25 years were randomly selected from three reservations in Minnesota and Wisconsin. There were 990 persons without diabetes, 294 with a prior diagnosis of diabetes, and 80 with fasting glucose >125 mg dl(-1) but no prior diabetes diagnosis. RESULTS: Persons with prior diabetes diagnosis were less likely than those without diabetes to report eating fast-food meals two or more times per week, eat visible fat on meat or the skin on poultry, eat fried chicken or fried fish, to add fat to cooked vegetables and drink whole milk. Persons with previously undiagnosed diabetes were more likely than previously diagnosed persons to report eating fast-food meals two or more times per week, eat visible fat on meat and the skin on poultry, drink whole milk and eat fried fish, but were less likely to drink low-fat milk. Previously undiagnosed persons were more likely than either diagnosed persons or persons without diabetes to consume lard from cooked foods and use it when cooking. CONCLUSION: Persons with diagnosed diabetes showed healthier eating patterns than those without diabetes, while undiagnosed persons showed some less favourable patterns. Because virtually all persons with diabetes in these communities receive nutrition education, the results suggest that nutrition education programmes for diabetics may be associated with healthier eating patterns. PMID- 15548341 TI - Motherhood meets epidemiology: measuring risk factors for breast-feeding cessation. AB - OBJECTIVE: To examine statistical models that have been used to predict the cessation of breast-feeding. SETTING: In nutritional epidemiology, a knowledge of risk factors that lead to breast-feeding cessation is essential to promote optimal infant health by increasing or sustaining breast-feeding rates. However, a number of methodological issues complicate the measurement of such risk factors. It is important when building multivariate models that variables entered into the model are not intervening variables, factors on the causal pathway or surrogate outcomes. Inclusion of these types of variable can lead to inaccurate models and biased results. A factor often cited to predict breast-feeding is 'intention to breast-feed' prior to the birth of the infant, although this factor is directly on the causal decision-making pathway. Another factor often cited is the age of introduction of formula feeding, which is actually part of the outcome variable because formula feeding defines the difference between full, complementary and no breast-feeding. Rather than include these as risk factors in multivariate models, factors removed from the causal pathway such as influences of educational practices, including advice to complementary feed, and beliefs and attitudes of families and health-care practitioners should be measured. CONCLUSIONS: The accurate quantification of modifiable risk factors is essential for designing public health education campaigns that are effective in sustaining or increasing breast-feeding duration. PMID- 15548342 TI - The cost of infant feeding in Liverpool, England. AB - OBJECTIVE: To investigate feeding practices in infants under the age of 4 months in Liverpool, England with particular reference to the cost of infant feeding. DESIGN: A cross-sectional survey consisting of self-completion questionnaires and interviews. SETTING: Subjects' homes within Central and South Liverpool Primary Care Trust areas. SUBJECTS: One hundred and forty-nine women (aged 18 to 43 years) and their infants (mean age 13 weeks). RESULTS: The average weekly cost of breast-feeding was 11.58 pounds sterling compared with 9.60 pounds sterling for formula-feeding. Many breast- and formula-feeding women spent money however on items that were not needed or used only once or twice. This was especially true of first-time mothers. Characteristics significantly associated with higher spending were: feeding method - mothers that had or were partially breast-feeding (P=0.001), education - those educated to degree level (P=0.028), socio-economic status - those in social classes I and II (P=0.002) and age - those aged 30 years and over (P=0.003). CONCLUSIONS: This study demonstrates that while breast feeding is often promoted as being free, this is not the case. Better information needs to be given to parents to avoid wasting money on items that are unnecessary, or where cheaper alternatives are available. PMID- 15548343 TI - Delivery of nutrition services in health systems in sub-Saharan Africa: opportunities in Burkina Faso, Mozambique and Niger. AB - BACKGROUND: In sub-Saharan Africa, underweight and micronutrient deficiencies account for an estimated 25% of the burden of disease. As the coverage of national health systems expands, increased opportunities exist to address the needs of children and women, the most vulnerable to these deficiencies, through high-quality nutrition services. OBJECTIVES: To assess health providers' knowledge and practice with regard to essential nutrition services for women and children in Burkina Faso, Mozambique and Niger, in order to assist the development of a standard guide and tools to assess and monitor the quality of the nutrition services delivered through national health systems. FINDINGS: The three surveys reveal the extent of missed opportunities to deliver nutrition services during routine prenatal, postnatal and child-care consultations for the prevention and treatment of highly prevalent nutritional deficiencies. CONCLUSION: A commitment to improving the quality of facility-based nutrition services is necessary to impact on the health outcomes of women and children 'covered' by national health systems. Rigorous assessment and monitoring of the quality of nutrition services should inform health programme and policy development. Building on the lessons learned in these three assessments, Helen Keller International has developed a standard Guide and Tools to assess the quality of the nutrition services delivered through national health systems. These tools can be adapted to assess ongoing nutrition services in health facilities, provide a framework for nutrition programming, inform the development of pre-service as well as in-service nutrition training curricula for providers, and evaluate the impact of nutrition training on providers' practices. PMID- 15548344 TI - The nutritional status of asymptomatic HIV-infected Africans: directions for dietary intervention? AB - OBJECTIVE: To compare the relationships between food (nutrient) intakes and biochemical markers of nutritional status of asymptomatic HIV-infected with HIV uninfected subjects, to gain more information on the appropriate diet for HIV infected persons at an early stage of infection. DESIGN: Cross-sectional population-based survey. SETTING: North West Province, South Africa. SUBJECTS: Two hundred and sixteen asymptomatic HIV-infected and 1550 HIV-uninfected men and women volunteers aged 15 years and older, recruited as 'apparently healthy' subjects from 37 randomly selected sites. OUTCOME MEASURES: Food and nutrient intakes, measured with a validated food-frequency questionnaire, and nutritional status indicated by anthropometric and biochemical variables, measured by a standardised methodology. RESULTS: The prevalence of HIV infection in the study population was 11.9%. The anthropometric indices and nutrient intakes of HIV infected and uninfected subjects did not differ significantly, indicating that these 216 HIV-infected subjects were at an early stage of infection. Of the biochemical nutritional status variables, high-density lipoprotein cholesterol and total cholesterol, haemoglobin, albumin and triglycerides were significantly lower in infected subjects. They also had higher globulin and liver enzyme levels than uninfected subjects. In infected subjects, serum albumin correlated significantly with serum lipids, serum vitamin A, serum vitamin E, serum iron, total iron-binding capacity and haemoglobin. The significant positive correlations of the liver enzymes with serum lipids, albumin, vitamin A and iron, observed in HIV-uninfected subjects, disappeared in the infected subjects. Polyunsaturated fat intake showed significant positive correlations with the increased liver enzymes in infected subjects. A principal components analysis indicated that, in infected subjects, increased liver enzymes correlated with higher consumption of maize meal and lower consumption of meat and vegetables. CONCLUSIONS AND RECOMMENDATIONS: This survey indicated that asymptomatic HIV infected subjects who followed a diet rich in animal foods had smaller decreases in serum albumin, haemoglobin and lipid variables, and smaller increases in liver enzymes, than those who consumed a diet based on staple foods. This suggests that animal foods are associated with improved nutritional status in HIV-infected persons. These results should be confirmed with intervention studies before dietary recommendations for asymptomatic HIV-infected individuals can be made. PMID- 15548345 TI - Anaemia and iron deficiency during pregnancy in rural Bangladesh. AB - OBJECTIVE: To study the prevalence of anaemia and its association with measures of iron deficiency (ID) among a group of pregnant women. DESIGN: Cross-sectional survey. SETTING: Pregnant women identified through house-to-house visits and participating in community-based antenatal care activities in a rural location of Mymensingh, Bangladesh. SUBJECTS: The estimates are based on 214 reportedly healthy pregnant women in their second trimester. Information on socio-economic status and reproductive history were obtained through home visits and venous blood samples were collected at antenatal care centres. Haemoglobin concentration (Hb) was measured by HemoCue, serum ferritin (sFt) by radioimmunoassay and serum transferrin receptor (sTfR) by enzyme-linked immunosorbent assay methods. ID was defined as presence of either low sFt (<12 microg l(-1)) or high sTfR (>8.5 mg l( 1)). RESULTS: The prevalence of anaemia (Hb <110 g l(-1)) was 50%, but severe anaemia (Hb <70 g l(-1)) was absent. Low sFt was observed in 42%, high sTfR in 25%, either low sFt or high TfR in 54% and both low sFt and high TfR in 13% of the pregnant women. Two out of three anaemic women had an indication of ID, which was present in 80% of women with moderate (Hb 70-99 g l(-1)) and 50% with mild (Hb 100-109 g l(-1)) anaemia. Four out of 10 non-anaemic women (Hb >/=110 g l( 1)) also had ID, but the prevalence was significantly lower than that observed in anaemic women (P=0.001). CONCLUSIONS: Despite the high prevalence of anaemia, severe cases were absent. The prevalence of ID increased at lower Hb. However, an increased prevalence was also found among women in the highest category of Hb. PMID- 15548346 TI - BRAC initiative towards promoting gender and social equity in health: a longitudinal study of child growth in Matlab, Bangladesh. AB - OBJECTIVE: To explore the effect of BRAC (formerly Bangladesh Rural Advancement Committee) initiatives towards promoting gender and social equity in health among children of poor mothers who are BRAC members. DESIGN: A cohort of 576 children from the prospective study of a BRAC- International Centre for Diarrhoeal Disease Research, Bangladesh joint research project was analysed. Data were collected three times during 1995-1996 with approximately 4-month intervals. Stunting, defined as height-for-age below minus two standard deviations from the reference median, was the outcome health measure. The study children were stratified into three groups according to their mother's social and BRAC membership status: poor and BRAC member (BM), poor non-member (TG) and non-poor non-member (NTG). SETTING: Matlab, rural area of Bangladesh. SUBJECTS: Children aged 6-72 months. RESULTS: The overall prevalence of stunting was 76%; the highest prevalence was found among TG (84.6%) children and no significant difference was observed between BM and NTG children (67.3% and 69.4%, respectively). In all groups, a significantly larger proportion of girls was stunted compared with boys in the first round. Group-level analysis showed that stunting decreased among all children except BM boys at the end of third round, with the largest decline among BM girls. In contrast, stunting prevalence increased among BM boys. A similar trend was found in the individual-level analysis, where a larger proportion of BM girls recovered from stunting compared with other groups and no recovery was observed among BM boys. At the end of the third round, the nutritional status of BM girls was almost equal to that of the BM boys, while gender inequity remained large among TG and NTG children. CONCLUSION: The BRAC initiative appeared to contribute to a significant equity gain in health for girls, as well as to decreased differences in ill health between the poor and the non-poor. PMID- 15548347 TI - Food store access and household fruit and vegetable use among participants in the US Food Stamp Program. AB - OBJECTIVE: Recent research on access to food among low-income populations in industrialised countries has begun to focus on neighbourhood food availability as a key determinant of dietary behaviour. This study examined the relationship between various measures of food store access and household fruit and vegetable use among participants in the Food Stamp Program, America's largest domestic food assistance programme. DESIGN: A secondary data analysis was conducted using the 1996-97 National Food Stamp Program Survey. The survey employed a 1-week food inventory method, including two at-home interviews, to determine household food use. Separate linear regression models were developed to analyse fruit and vegetable use. Independent variables included distance to store, travel time to store, ownership of a car and difficulty of supermarket access. All models controlled for a full set of socio-economic variables. SUBJECTS: A nationally representative sample of participants (n=963) in the Food Stamp Program. RESULTS: After controlling for confounding variables, easy access to supermarket shopping was associated with increased household use of fruits (84 grams per adult equivalent per day; 95% confidence interval 5, 162). Distance from home to food store was inversely associated with fruit use by households. Similar patterns were seen with vegetable use, though associations were not significant. CONCLUSIONS: Environmental factors are importantly related to dietary choice in a nationally representative sample of low-income households, reinforcing the importance of including such factors in interventions that seek to effect dietary improvements. PMID- 15548348 TI - Eating at fast-food restaurants is associated with dietary intake, demographic, psychosocial and behavioural factors among African Americans in North Carolina. AB - OBJECTIVE: To examine associations of the frequency of eating at fast-food restaurants with demographic, behavioural and psychosocial factors and dietary intake in African American adults. METHODS: Self-reported data from a population based cross-sectional survey of 658 African Americans, aged 20-70 years, in North Carolina. An 11-page questionnaire assessed eating at fast-food restaurants, demographic, behavioural and diet-related psychosocial factors, and dietary intake (fruit, vegetable, total fat and saturated fat intakes, and fat-related dietary behaviours). RESULTS: The participants were aged 43.9+/-11.6 years (mean+/-standard deviation), 41% were male, 37% were college graduates and 75% were overweight or obese. Seventy-six per cent reported eating at fast-food restaurants during the previous 3 months: 4% usually, 22% often and 50% sometimes. Frequency of eating at fast-food restaurants was positively associated with total fat and saturated fat intakes and fat-related dietary behaviours (P<0.0001) and inversely associated with vegetable intake (P<0.05). For example, mean daily fat intake was 39.0 g for usually/often respondents and 28.3 g for those reporting rare/never eating at fast-food restaurants. Participants who reported usual/often eating at fast-food restaurants were younger, never married, obese, physically inactive and multivitamin non-users (all P<0.01). Frequency of eating at fast-food restaurants was positively associated with fair/poor self rated health, weak belief in a diet-cancer relationship, low self-efficacy for healthy eating, weight dissatisfaction, and perceived difficulties of preparing healthy meals and ordering healthy foods in restaurants (all P<0.05). Frequency of eating at fast-food restaurants did not differ significantly by sex, education, smoking, ability to purchase healthy foods or knowledge of the Food Guide Pyramid. CONCLUSIONS: Eating at fast-food restaurants is associated with higher fat and lower vegetable intakes in African Americans. Interventions to reduce fast-food consumption and obesity in African Americans should consider demographic and behavioural characteristics and address attitudes about diet disease relationships and convenience barriers to healthy eating. PMID- 15548349 TI - Performance of a short tool to assess dietary intakes of fruits and vegetables, percentage energy from fat and fibre. AB - OBJECTIVES: We describe the methods used to develop and score a 17-item 'screener' designed to estimate intake of fruit and vegetables, percentage energy from fat and fibre. The ability of this screener and a food-frequency questionnaire (FFQ) to measure these exposures is evaluated. DESIGN: Using US national food consumption data, stepwise multiple regression was used to identify the foods to be included on the instrument; multiple regression analysis was used to develop scoring algorithms. The performance of the screener was evaluated in three different studies. Estimates of intakes measured by the screener and the FFQ were compared with true usual intake based on a measurement error model. SETTING: US adult population. SUBJECTS: For development of instrument, n=9323 adults. For testing of instrument, adult men and women in three studies completing multiple 24-hour dietary recalls, FFQ and screeners, n=484, 462 and 416, respectively. RESULTS: Median recalled intakes for examined exposures were generally estimated closely by the screener. In the various validation studies, the correlations between screener estimates and estimated true intake were 0.5 0.8. In general, the performances of the screener and the full FFQ were similar; estimates of attenuation were lower for screeners than for full FFQs. CONCLUSIONS: When coupled with appropriate reference data, the screener approach described may yield useful estimates of intake, for both surveillance and epidemiological purposes. PMID- 15548350 TI - Chemotherapy-induced and/or radiation therapy-induced oral mucositis- complicating the treatment of cancer. AB - The term mucositis is coined to describe the adverse effects of radiation and chemotherapy treatments. Mucositis is one of the most common adverse reactions encountered in radiation therapy for head and neck cancers, as well as in chemotherapy, in particular with drugs affecting DNA synthesis (S-phase-specific agents such as fluorouracil, methotrexate, and cytarabine). Mucositis may limit the patient's ability to tolerate chemotherapy or radiation therapy, and nutritional status is compromised. It may drastically affect cancer treatment as well as the patient's quality of life. The incidence and severity of mucositis will vary from patient to patient. It will also vary from treatment to treatment. It is estimated that there is 40% incidence of mucositis in patients treated with standard chemotherapy and this will not only increase with the number of treatment cycles but also with previous episodes. Similarly, patients who undergo bone marrow transplantation and who receive high doses of chemotherapy have a 76% chance of getting mucositis. Patients receiving radiation, in particular to head and neck cancers, have a 30% to 60% chance. The exact pathophysiology of development is not known, but it is thought to be divided into direct and indirect mucositis. Chemotherapy and/or radiation therapy will interfere with the normal turnover of epithelial, cells leading to mucosal injury; subsequently, it can also occur due to indirect invasion of Gram-negative bacteria and fungal species because most of the cancer drugs will cause changes in blood counts. With the advancement in cytology, a more precise mechanism has been established. With this understanding, we can select and target particular mediators responsible for the mucositis. Risk factors such as age, nutritional status, type of malignancy, and oral care during treatment will play important roles in the development of mucositis. Many treatment options are available to prevent and treat this condition, but none of them can completely prevent or treat mucositis. More and more pathological methods are being developed to understand this condition so that better therapeutic regimens can be selected. Emphasis also should be made in assessing the patient's psychologic condition, particular depressive disorders. This is important because treatment with antidepressants will not only contribute in lifting depression but also reduces pain somatization. Although mucositis is rarely life-threatening, it will interfere with treatment of cancer to a great extent. PMID- 15548351 TI - High-resolution genomic and expression profiling reveals 105 putative amplification target genes in pancreatic cancer. AB - Comparative genomic hybridization (CGH) studies have provided a wealth of information on common copy number aberrations in pancreatic cancer, but the genes affected by these aberrations are largely unknown. To identify putative amplification target genes in pancreatic cancer, we performed a parallel copy number and expression survey in 13 pancreatic cancer cell lines using a 12,232 clone cDNA microarray, providing an average resolution of 300 kb throughout the human genome. CGH on cDNA microarray allowed highly accurate mapping of copy number increases and resulted in identification of 24 independent amplicons, ranging in size from 130 kb to 11 Mb. Statistical evaluation of gene copy number and expression data across all 13 cell lines revealed a set of 105 genes whose elevated expression levels were directly attributable to increased copy number. These included genes previously reported to be amplified in cancer as well as several novel targets for copy number alterations, such as p21-activated kinase 4 (PAK4), which was previously shown to be involved in cell migration, cell adhesion, and anchorage-independent growth. In conclusion, our results implicate a set of 105 genes that is likely to be actively involved in the development and progression of pancreatic cancer. PMID- 15548352 TI - Transcriptional repression of catalase in mouse skin tumor progression. AB - Previous studies in our laboratory have shown that the elevation of reactive oxygen species levels and the repression of the antioxidant enzyme, catalase, played a critical role in the in vitro progression of benign papilloma cells to malignant carcinoma cells. Catalase message, protein levels, and activity levels were found to be downregulated in the malignantly progressed cells. The goal of this study is to further characterize the repression of catalase in malignant progression of mouse skin tumors. To validate the in vitro observations, we examined catalase expression in tumor samples generated by the multistep chemical carcinogenesis protocol. Higher levels of catalase mRNA and protein were observed in benign papillomas versus malignant carcinomas. Nuclear run-on analysis showed that catalase repression in the cultured malignant cells was transcription dependent. Results from luciferase reporter assays indicated that malignant cells have lower catalase promoter activities than benign papilloma cells, in part through the Wilm's tumor suppressor 1 (WT1) binding site within the proximal promoter region. The WT1 protein levels were found to be inversely correlated with the observed catalase promoter activities, with higher levels observed in the malignant cells versus the benign cells. These results led us to conclude that WT1 is acting as a transcription repressor in catalase gene regulation during tumor progression. PMID- 15548353 TI - Validity of serum pepsinogen I/II ratio for the diagnosis of gastric epithelial dysplasia and intestinal metaplasia during the follow-up of patients at risk for intestinal-type gastric adenocarcinoma. AB - A cohort of individuals (n = 136) with lesions as severe as atrophic chronic gastritis (ACG) was cross-sectionally evaluated for the validity assessment of pepsinogen I (PGI) and pepsinogen II (PGII) serum levels for the diagnosis of intestinal metaplasia (IM) and gastric dysplasia. PGI/PGII ratio [median (range)] was 4 (0.5-7.5) in patients with ACG (n = 35); 4.6 (1.9-6.8) in type I IM (n = 18); 4.2 (1.4-5.9) in type II or type III IM limited to the antrum and incisura (n = 20); 2.4 (0.4-5.6) in extensive incomplete IM (n = 38); and 1.3 (0.4-6.4) in low-grade dysplasia (n = 23) (P = .002). Using histopathologic data as a reference test, the area under the receiver operating characteristic curves (CI 95%) was 0.73 (0.64-0.82) for extensive IM, 0.72 (0.58-0.85) for the diagnosis of dysplasia, and 0.81 (0.66-0.95) for the diagnosis of high-grade dysplasia. Using a PGI/PGII ratio of < or =3 as the cutoff for dysplasia diagnosis, the sensitivity was 70% (62-78%), the specificity was 65% (57-73%), and the negative predictive value estimates were over 90%. No differences in PG levels according to age or gender were observed. Helicobacter pylori did not significantly influence validity measurement estimates. PGI/PGII serum level ratio can be used even in the management of patients with a high a priori probability for a positive test. It may be useful for the exclusion of more advanced lesions (extensive IM and neoplastic lesions). PMID- 15548354 TI - The novel highly lipophilic topoisomerase I inhibitor DB67 is effective in the treatment of liver metastases of murine CT-26 colon carcinoma. AB - Colorectal carcinoma occurs in 1 of 20 individuals in most developed countries. The relapse after resection with metastatic liver disease is a major cause of death. 7-t-Butyldimethylsilyl-10-hydroxycamptothecin (DB67) has been incorporated into liposomes allowing for intravenous (i.v.) administration. A preclinical efficacy study of liposomal DB67 was performed using the colon carcinoma CT-26 cell line. The therapeutic dose for DB67 and liposomal DB67 was found to be 7 mg/kg per day using the qdx5/1 schedule. The results are compared with those obtained with irinotecan. The treatment with liposomal DB67 administered intravenously was more effective in reducing the weight and volume of primary spleen tumors and the weight and extent of liver metastases than free DB67 or liposomal DB67 administered intraperitoneally, but less effective than irinotecan. When the primary tumor was resected, treatment with liposomal DB67 administered intravenously was more effective in reducing the weight and extent of liver metastases than DB67 or liposomal DB67 administered intraperitoneally, and irinotecan. DB67 showed a higher accumulation in spleen and liver after its i.v. administration in liposomal form compared with its free or liposomal form administered intraperitoneally. DB67 and liposomal DB67 are more effective than irinotecan in the treatment of liver metastases after resection of the primary tumor. PMID- 15548355 TI - Laminin-5 beta3A expression in LNCaP human prostate carcinoma cells increases cell migration and tumorigenicity. AB - Interactions between extracellular matrix proteins and prostate carcinoma cells change dramatically during prostate tumor progression. We have concentrated on two key modifications that occur in the hemidesmosome in prostate carcinoma: loss of laminin-5 protein expression and altered basal cell polarity of the alpha6beta4 integrin. We previously demonstrated two cell line-specific isoforms (beta3A and beta3B) of the LAMB3 message. Cells expressing only the beta3B isoform did not translate the beta3 protein and were unable to assemble the laminin-5 trimer. One such cell line, LNCaP, was selected to determine whether restoration of the laminin-5 beta3A isoform would cause expression of a functional laminin-5 beta3 chain, assembly and secretion of the laminin-5 trimer, and reversion to a non-neoplastic phenotype. Laminin-5 beta3A cDNA was cloned and stably transfected into LNCaP cells. We observed the restoration of the beta3 protein, but a laminin-5 trimer was not secreted. Moreover, increased cell migration was demonstrated, and tumorigenicity was increased in SCID mice. A microarray analysis, performed between transfected and nontransfected LNCaP cells, showed most changing genes to be associated with signal transduction. The beta3 chain of laminin-5 may thus play an important role in signal transduction, which may enhance cell motility and tumorigenesis. PMID- 15548356 TI - Halofuginone inhibits angiogenesis and growth in implanted metastatic rat brain tumor model--an MRI study. AB - Tumor growth and metastasis depend on angiogenesis; therefore, efforts are made to develop specific angiogenic inhibitors. Halofuginone (HF) is a potent inhibitor of collagen type alpha1(I). In solid tumor models, HF has a potent antitumor and antiangiogenic effect in vivo, but its effect on brain tumors has not yet been evaluated. By employing magnetic resonance imaging (MRI), we monitored the effect of HF on tumor progression and vascularization by utilizing an implanted malignant fibrous histiocytoma metastatic rat brain tumor model. Here we demonstrate that treatment with HF effectively and dose-dependently reduced tumor growth and angiogenesis. On day 13, HF-treated tumors were fivefold smaller than control (P < .001). Treatment with HF significantly prolonged survival of treated animals (142%; P = .001). In HF-treated rats, tumor vascularization was inhibited by 30% on day 13 and by 37% on day 19 (P < .05). Additionally, HF treatment inhibited vessel maturation (P = .03). Finally, in HF treated rats, we noticed the appearance of a few clusters of satellite tumors, which were distinct from the primary tumor and usually contained vessel cores. This phenomenon was relatively moderate when compared to previous reports of other antiangiogenic agents used to treat brain tumors. We therefore conclude that HF is effective for treatment of metastatic brain tumors. PMID- 15548357 TI - Gene expression profiling of mouse teratocarcinomas uncovers epigenetic changes associated with the transformation of mouse embryonic stem cells. AB - The molecular mechanisms of the development of teratocarcinomas from stem cells are largely unknown. To determine which genes are associated with the transformation of these cells, we have performed oligonucleotide microarray analysis, using Affymetrix U74A GeneChips, on both cell cultures and tumors in nude mice. We identified 68 genes that significantly differed in expression between the ES cell culture and the teratocarcinoma cell line, SCC-PSA1, and 51 genes with statistically different expression patterns between the ES cell tumors and the teratocarcinomas (P < .00005). We found that there were 20 genes that had common expression patterns in both groups. We also examined the role of the transition from in vitro to in vivo by comparing ES cell culture to ES cell tumor, and teratocarcinoma cell line to teratocarcinomas. We identified 22 genes that were upregulated in the ES cell tumors and 42 that had a decreased expression in the tumor (P < .0001). In comparing SCC-PSA1 to its tumor, we identified 34 upregulated genes and 25 downregulated genes (P < .001). There were only 10 genes in common from these two lists. GenMapp search revealed that several pathways, especially the cell cycle pathway, are actively involved in the induction of teratocarcinomas. Our results indicate that many key development genes may play a key role in the transformation of ES cells into teratocarcinoma cells. PMID- 15548358 TI - Expression of the platelet-derived growth factor receptor in prostate cancer and treatment implications with tyrosine kinase inhibitors. AB - The platelet-derived growth factor receptor (PDGFR) is a receptor tyrosine kinase overexpressed in a subset of solid tumors and therefore is the target of drugs inhibiting this function such as imatinib mesylate (Gleevec). Thus far, drug therapy has played a limited role in the treatment of localized prostate cancer (PCa). This study characterizes PDGFR-beta expression in a wide spectrum of PCa samples to provide empirical data as part of a rational treatment strategy. A survey of five published prostate expression array studies, including 100 clinically localized PCa, did not identify tumors with increased PDGFR-beta expression level. Protein expression of PDGFR-beta, as determined by immunohistochemistry, revealed 5% of clinically localized PCa and 16% of metastatic PCa cases to show moderate or strong expression. To develop a strategy to detect patients most likely to profit from Gleevec treatment, we analyzed cDNA expression array data from 10,000 transcripts for PDGFR-beta expression and divided tumors in groups based on PDGFR-beta expression level. Performing a supervised analysis to identify potential comarkers of PDGFR-beta in PCa, we identified a set of genes whose expression was associated with PDGFR-beta status including early growth response 1 (Egr1), an upstream effector of PDGF (4.2-fold upregulation), alpha-methylacyl-CoA racemase, as well as v-Maf and neuroblastoma suppressor of tumorigenicity (both with a 2.2-fold downregulation). Taken together, this study suggests that only a small subset of PCas may be amenable to tyrosine kinase inhibitors specific for PDGFR. PMID- 15548359 TI - Lonidamine causes inhibition of angiogenesis-related endothelial cell functions. AB - The aim of this study was to assess whether lonidamine (LND) interferes with some steps in angiogenesis progression. We report here, for the first time, that LND inhibited angiogenic-related endothelial cell functions in a dose-dependent manner (1-50 microg/ml). In particular, LND decreased proliferation, migration, invasion, and morphogenesis on matrigel of different endothelial cell lines. Zymographic and Western blot analysis assays showed that LND treatment produced a reduction in the secretion of matrix metalloproteinase-2 and metalloproteinase-9 by endothelial cells. Vessel formation in a matrigel plug was also reduced by LND. The viability, migration, invasion, and matrix metalloproteinase production of different tumor cell lines were not affected by low doses of LND (1-10 microg/ml), whereas 50 microg/ml LND, which corresponds to the dose used in clinical management of tumors, triggered apoptosis both in endothelial and tumor cells. Together, these data demonstrate that LND is a compound that interferes with endothelial cell functions, both at low and high doses. Thus, the effect of LND on endothelial cell functions, previously undescribed, may be a significant contributor to the antitumor effect of LND observed for clinical management of solid tumors. PMID- 15548360 TI - Cytokines and chemokines are expressed at different levels in small and large murine colon-26 tumors following intratumoral injections of CpG ODN. AB - Direct tumor injections of (CpG ODN) into murine colon tumor 26 (CT-26) tumors can induce a potent antitumor response. Tumor size at the beginning of treatment determines the final therapeutic outcome, with smaller tumors responding favorably to CpG ODN therapy whereas large tumors do not. CpG ODN injections in small tumors resulted in tumor necrosis and extensive inflammatory cell infiltration, with average survival that is significantly higher (48.1 +/- 34 days) when compared to control ODN-treated mice (16.1 +/- 3.5 days). Cytokines and chemokines are expressed at different levels in small and large CT-26 tumors following intratumoral injections of CpG ODN. We observed that granulocyte macrophage colony-stimulating factor and interleukin (IL) 6 are the major cytokines that were overexpressed in CpG ODN-treated small tumors but not in large tumors. Similarly, several chemokines (CXCL1, CCL2, and CCL3) were also significantly higher in CpG ODN-treated small tumors compared to control ODN treated tumors. PMID- 15548361 TI - Selective loss of codon 72 proline p53 and frequent mutational inactivation of the retained arginine allele in colorectal cancer. AB - According to recent reports, some cancer types exhibit nonrandom allele loss at codon 72 in exon 4 of the p53 gene [coding for proline (72Pro) or arginine (72Arg)]. To clarify this phenomenon for colorectal cancer and to find out if this preferential loss might have any functional significance, p53 loss of heterozygosity (LOH) and p53 mutations were investigated in a group of 61 colorectal cancers and 28 liver metastases, and were correlated with clinicopathologic factors. A comparison of a patient's blood codon 72 status with a healthy control group did not reveal an enhanced risk of developing colorectal tumors for one of the two isoforms. p53-LOH and p53 mutations were found in 62.2% and 39.4% of primary tumors, respectively, and in 57.9% and 25% of hepatic metastases, respectively. In 14 heterozygous cases showing exon 4-LOH, only the 72Pro allele was lost and the retained 72Arg was preferentially mutated. In general, p53 mutations were significantly associated with the 72Arg tumor status (P < .001). Distal tumors showed allelic losses of the p53 gene more commonly than proximal tumors (P = .054). The prevalence of 72Arg increased in frequency with higher Dukes stage (P = .056). We suggest that either the preferential loss of 72Pro or the mutation of the 72Arg in colorectal cancer and hepatic metastases is associated with malignant potential and might reflect carcinogenic exposure, particularly in the distal part of the large intestines. PMID- 15548362 TI - Detection of colorectal cancer by a quantitative fluorescence determination of DNA amplification in stool. AB - DNA amplification of exfoliated cells in stool represents an inexpensive and rapid test, but has only 50% to 60% sensitivity. A new quantitative method, called fluorescence long DNA, was developed and validated in our laboratory on stool obtained from 86 patients with primary colorectal cancer and from 62 healthy individuals. It consists of the amplification of stool DNA with fluorescence primers and the quantification of the amplification using a standard curve. Results are arbitrarily expressed in nanograms. The potential of the new method compared to the conventional approach was analyzed in a subgroup of 94 individuals (56 patients and 38 healthy volunteers). In the present series, DNA amplification analysis showed a specificity of 97% and a sensitivity of only 50%. Conversely, fluorescence DNA evaluation, using the best cutoff of 25 ng, showed a sensitivity of about 76% and a specificity of 93%. Similar sensitivity was observed regardless of Dukes stage, tumor location, and size, thus also permitting the detection of early-stage tumors. The present study seems to indicate that quantitative fluorescence DNA determination in stool successfully identifies colorectal cancer patients with a sensitivity comparable, if not superior, to that of multiple gene analysis but at a lower cost and in a shorter time. PMID- 15548363 TI - BRCA2 mutations in 154 finnish male breast cancer patients. AB - The etiology and pathogenesis of male breast cancer (MBC) are poorly known. This is due to the fact that the disease is rare, and large-scale genetic epidemiologic studies have been difficult to carry out. Here, we studied the frequency of eight recurrent Finnish BRCA2 founder mutations in a large cohort of 154 MBC patients (65% diagnosed in Finland from 1967 to 1996). Founder mutations were detected in 10 patients (6.5%), eight of whom carried the 9346(-2) A>G mutation. Two novel mutations (4075 delGT and 5808 del5) were discovered in a screening of the entire BRCA2 coding region in 34 samples. However, these mutations were not found in the rest of the 120 patients studied. Patients with positive family history of breast and/or ovarian cancer were often BRCA2 mutation carriers (44%), whereas those with no family history showed a low frequency of involvement (3.6%; P < .0001). Finally, we found only one Finnish MBC patient with 999 del5, the most common founder mutation in Finnish female breast cancer (FBC) patients, and one that explains most of the hereditary FBC and MBC cases in Iceland. The variation in BRCA2 mutation spectrum between Finnish MBC patients and FBC patients in Finland and breast cancer patients in Iceland suggests that modifying genetic and environmental factors may significantly influence the penetrance of MBC and FBC in individuals carrying germline BRCA2 mutations in some populations. PMID- 15548364 TI - Multiple stress signals induce p73beta accumulation. AB - Although p73 is a structural and functional homologue of the tumor-suppressor gene p53, it is not mutated in many human cancers as p53. Besides, p73 was shown to be activated by only a subset of signals that activate p53, such as gamma irradiation and cisplatin, but not by other common genotoxic stress-inducing agents such as ultraviolet (UV) irradiation, although many of these signals are also capable of inducing p53-independent cell death. Using a p73-specific antibody, we confirmed that c-Abl is required for cisplatin-induced p73 upregulation, and further demonstrate that the p73 protein is upregulated by UV irradiation and other stress stimuli including sorbitol, hydrogen peroxide, nocodazol, and taxol. These stress signals upregulate both p73 mRNA and increases the stability of p73, indicating that p73 is regulated transcriptionally and posttranslationally. Cells stably expressing the dominant-negative p73 inhibitor protein (p73DD) and p73(-/-) fibroblasts are more resistant than control cells to apoptosis induced by these stress signals, suggesting that p73 contributes to apoptosis induction. Together, the data demonstrate that several stress signals can signal to p73 in vivo, which raises the possibility of eradicating cancers with an unmutated p73 gene by activating them with stress-inducing agents or their mimetics. PMID- 15548365 TI - Downregulation and/or release of NKG2D ligands as immune evasion strategy of human neuroblastoma. AB - Neuroblastoma (NB) is a pediatric extracranial tumor characterized by downregulation of human leukocyte antigen class I and defects of the antigen processing machinery, two features that make it an appropriate target for natural killer (NK)-mediated lysis. NKG2D is an activating immunoreceptor expressed by cytotoxic T lymphocytes and NK cells. The ligands for NKG2D are the major histocompatibility complex class I-related chain (MIC)A and MICB glycoproteins, and the UL-16-binding proteins (ULBPs). Here, the expression of NKG2D ligands was investigated in human primary NB tumors and cell lines because scanty information is available on this issue. MICA, MICB, and ULBP transcripts were found in most tumors and cell lines. MICA protein was detected in some NB cell lines but not in primary tumors. A soluble form of MICA (sMICA) was identified in most patient sera and in some cell line supernatants. sMICA downregulated surface NKG2D in normal peripheral blood CD8(+) cells and decreased NK-mediated killing of MICA(+) NB cells. MICB was detected exclusively in the cytosol of primary tumors and cell lines. Approximately 50% of primary tumors expressed ULBP-2, but not ULBP-1 or 3. ULBP-3 was expressed in 5 of 9 cell lines, ULBP-2 in 2 of 9, whereas ULBP-1 was never detected. These studies delineate novel potential pathways of tumor escape and immunodeficiency in NB. PMID- 15548366 TI - Altered gene expression profile in mouse bladder cancers induced by hydroxybutyl(butyl)nitrosamine. AB - A variety of genetic alterations and gene expression changes are involved in the pathogenesis of bladder tumor. To explore these changes, oligonucleotide array analysis was performed on RNA obtained from carcinogen-induced mouse bladder tumors and normal mouse bladder epithelia using Affymetrix (Santa Clara, CA) MGU74Av2 GeneChips. Analysis yielded 1164 known genes that were changed in the tumors. Certain of the upregulated genes included EGFR-Ras signaling genes, transcription factors, cell cycle-related genes, and intracellular signaling cascade genes. However, downregulated genes include mitogen-activated protein kinases, cell cycle checkpoint genes, Rab subfamily genes, Rho subfamily genes, and SH2 and SH3 domains-related genes. These genes are involved in a broad range of different pathways including control of cell proliferation, differentiation, cell cycle, signal transduction, and apoptosis. Using the pathway visualization tool GenMAPP, we found that several genes, including TbR-I, STAT1, Smad1, Smad2, Jun, NFkappaB, and so on, in the TGF-beta signaling pathway and p115 RhoGEF, RhoGDI3, MEKK4A/MEKK4B, PI3KA, and JNK in the G13 signaling pathway were differentially expressed in the tumors. In summary, we have determined the expression profiles of genes differentially expressed during mouse bladder tumorigenesis. Our results suggest that activation of the EGFR-Ras pathway, uncontrolled cell cycle, aberrant transcription factors, and G13 and TGF-beta pathways are involved, and the cross-talk between these pathways seems to play important roles in mouse bladder tumorigenesis. PMID- 15548367 TI - Molecular cytogenetic characterization of tenosynovial giant cell tumors. AB - Tenosynovial giant cell tumor (TSGCT) is a disease of disputed etiology and pathogenesis. Some investigations indicate a neoplastic origin of the tumors; others indicate that they are polyclonal and inflammatory. The cytogenetic and molecular genetic features of TSGCTs are largely unknown, as only some 20 localized and 30 diffuse tumors with cytogenetic aberrations have been reported. The most common karyotypic aberrations have been trisomy for chromosomes 5 and 7 and translocations involving chromosomal area 1p11-13. We decided to screen the genomes of TSGCTs by comparative genomic hybridization (CGH) to perform interphase fluorescence in situ hybridization (IP-FISH), looking for numerical aberrations of chromosomes 1, 5, and 7, and to analyze the tumors for microsatellite instability. Except for two diffuse TSGCTs that came fresh to us, and which, by karyotyping, exhibited t(1;22)(p13;q12) and a t(1;1)(q21;p11) and +7, respectively, all studies had to be performed on formalin-fixed, paraffin embedded material. DNA was extracted from 51 localized and nine diffuse TSGCTs. CGH was successful for 24 tumors, but none of them showed copy number changes. The IP-FISH studies showed trisomy 7 in 56% of the tumors (15/27), whereas chromosomes 1 and 5 seemed to be disomic in all TSGCTs. All informative tumors were wild-type by microsatellite instability analysis. PMID- 15548368 TI - Developmental effects of prenatal exposure to bisphenol a on the uterus of rat offspring. AB - Exposure to estrogenic compounds during critical periods of fetal development could result in adverse effects on the development of reproductive organs that are not apparent until later in life. Bisphenol A (BPA), which is employed in the manufacture of a wide range of consumer products, is a prime candidate for endocrine disruption. We examined BPA to address the question of whether in utero exposure affects the uterus of the offspring and studied the expression and distribution of the estrogen receptors alpha (ERalpha) and beta (ERbeta), because estrogens influence the development, growth, and function of the uterus through both receptors. Gravid Sprague-Dawley dams were administered by gavage either 0.1 or 50 mg/kg per day BPA or 0.2 mg/kg per day 17alpha-ethinyl estradiol (EE2) as reference dose on gestation days 6 through 21. Female offspring were killed in estrus. Uterine morphologic changes as well as ERalpha and ERbeta distribution and expression were measured by immunohistochemistry and Western blot analysis. Striking morphologic changes were observed in the uterine epithelium of postpubertal offspring during estrus of the in utero BPA-treated animals (the thickness of the total epithelium was significantly reduced). ERalpha expression was increased in the 50-mg BPA and EE2-treated group. In contrast, we observed significantly decreased ERbeta expression in all BPA- and EE2-treated animals when compared with the control. In summary, these results clearly indicate that in utero exposure of rats to BPA promotes uterine disruption in offspring. We hypothesize that the uterine disruption could possibly be provoked by a dysregulation of ERalpha and ERbeta. PMID- 15548369 TI - Differential regulation of a fibroblast growth factor-binding protein during skin carcinogenesis and wound healing. AB - The initiation of premalignant lesions is associated with subtle cellular and gene expression changes. Here we describe a severe combined immunodeficiency mouse xenograft model with human adult skin and compare chemical carcinogenesis and wound healing. We focus on a secreted binding protein for fibroblast growth factors (FGF-BP) that enhances the activity of locally stored FGFs and is expressed at high levels in human epithelial cancers. Carcinogen treatment of murine skin induced papilloma within 6 weeks, whereas the human skin grafts displayed no obvious macroscopic alterations. Microscopic studies of the human skin, however, showed p53-positive keratinocytes in the epidermis, increased angiogenesis in the dermis of the treated skin, enhanced proliferation of keratinocytes in the basal layer, and an increase of FGF-BP protein and mRNA expression. In contrast, after surgical wounding of human skin grafts or of mouse skin, FGF-BP expression was upregulated within a few hours and returned to control levels after 2 days with wound closure. Enhanced motility of cultured keratinocytes and dermal fibroblasts by FGF-BP supports a role in wound healing. We conclude that adult human skin xenografts can be used to identify early molecular events during malignant transformation as well as transient changes during wound healing. PMID- 15548370 TI - Activation of the Erk pathway is required for TGF-beta1-induced EMT in vitro. AB - Transforming growth factor-beta1 (TGF-beta1) can be tumor-suppressive through the activation of the Smad-mediated signaling pathway. TGF-beta1 can also enhance tumor progression by stimulating epithelial-to-mesenchymal transition (EMT) through additional pathways. EMT is characterized by the acquisition of a fibroblast-like cell morphology, dissolution of tight junctions, disruption of adherence junctions, and formation of actin stress fibers. There is evidence linking the activation of mitogen-activated protein kinase pathways to the induction of TGF-beta1-mediated EMT. However, the role of Erk in the induction of TGF-beta1-mediated EMT remains unclear. TGF-beta1 treatment of normal murine mammary gland (NMuMG) epithelial cells resulted in increased gene expression of Ras, Raf, MEK1/2, and Erk1/2, as shown by microarray analysis and real-time polymerase chain reaction. Upon 24 and 48 hours of treatment with TGF-beta1, NMuMG and mouse cortical tubule (MCT) epithelial cells underwent EMT as shown by changes in cell morphology, delocalization of zonula occludens-1 and E-cadherin from cell-cell junctions, and formation of actin stress fibers. TGF-beta1 treatment also resulted in increased levels of phosphorylated Erk and Erk kinase activity. Treatment with an MEK inhibitor, U0126, inhibited increased Erk phosphorylation and kinase activity, and blocked TGF-beta1-induced EMT in both cell lines. These data show that TGF-beta1 induces the activation of the Erk signaling pathway, which is required for TGF-beta1-mediated EMT in vitro. PMID- 15548371 TI - Gene expression profiling of microdissected pancreatic ductal carcinomas using high-density DNA microarrays. AB - Pancreatic ductal adenocarcinoma (PDAC) remains an important cause of malignancy related death and is the eighth most common cancer with the lowest overall 5-year relative survival rate. To identify new molecular markers and candidates for new therapeutic regimens, we investigated the gene expression profile of microdissected cells from 11 normal pancreatic ducts, 14 samples of PDAC, and 4 well-characterized pancreatic cancer cell lines using the Affymetrix U133 GeneChip set. RNA was extracted from microdissected samples and cell lines, amplified, and labeled using a repetitive in vitro transcription protocol. Differentially expressed genes were identified using the significance analysis of microarrays program. We found 616 differentially expressed genes. Within these, 140 were also identified in PDAC by others, such as Galectin-1, Galectin-3, and MT-SP2. We validated the differential expression of several genes (e.g., CENPF, MCM2, MCM7, RAMP, IRAK1, and PTTG1) in PDAC by immunohistochemistry and reverse transcription polymerase chain reaction. We present a whole genome expression study of microdissected tissues from PDAC, from microdissected normal ductal pancreatic cells and pancreatic cancer cell lines using high-density microarrays. Within the panel of genes, we identified novel differentially expressed genes, which have not been associated with the pathogenesis of PDAC before. PMID- 15548372 TI - Genomic profiles in stage I primary non small cell lung cancer using comparative genomic hybridization analysis of cDNA microarrays. AB - To investigate the genomic aberrations that are involved in lung tumorigenesis and therefore may be developed as biomarkers for lung cancer diagnosis, we characterized the genomic copy number changes associated with individual genes in 14 tumors from patients with primary non small cell lung cancer (NSCLC). Six squamous cell carcinomas (SQCAs) and eight adenocarcinomas (ADCAs) were examined by high-resolution comparative genomic hybridization (CGH) analysis of cDNA microarray. The SQCAs and ADCAs shared common frequency distributions of recurrent genomic gains of 63 genes and losses of 72 genes. Cluster analysis using 57 genes defined the genomic differences between these two major histologic types of NSCLC. Genomic aberrations from a set of 18 genes showed distinct difference of primary ADCAs from their paired normal lung tissues. The genomic copy number of four genes was validated by fluorescence in situ hybridization of 32 primary NSCLC tumors, including those used for cDNA microarray CGH analysis; a strong correlation with cDNA microarray CGH data emerged. The identified genomic aberrations may be involved in the initiation and progression of lung tumorigenesis and, most importantly, may be developed as new biomarkers for the early detection and classification of lung cancer. PMID- 15548373 TI - Exceptionally potent anti-tumor bystander activity of an scFv:sTRAIL fusion protein with specificity for EGP2 toward target antigen-negative tumor cells. AB - Previously, we reported on the target cell-restricted fratricide apoptotic activity of scFvC54:sTRAIL, a fusion protein comprising human-soluble tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) genetically linked to the antibody fragment scFvC54 specific for the cell surface target antigen EGP2. In the present study, we report that the selective binding of scFvC54:sTRAIL to EGP2-positive target cells conveys an exceptionally potent pro-apoptotic effect toward neighboring tumor cells that are devoid of EGP2 expression (bystander cells). The anti-tumor bystander activity of scFvC54:sTRAIL was detectable at target-to-bystander cell ratios as low as 1:100. Treatment in the presence of EGP2-blocking or TRAIL-neutralizing antibody strongly inhibited apoptosis in both target and bystander tumor cells. In the absence of target cells, bystander cell apoptosis induction was abrogated. The bystander apoptosis activity of scFvC54:sTRAIL did not require internalization, enzymatic conversion, diffusion, or communication (gap junctional intracellular communication) between target and bystander cells. Furthermore, scFvC54:sTRAIL showed no detectable signs of innocent bystander activity toward freshly isolated blood cells. Further development of this new principle is warranted for approaches where cancer cells can escape from antibody-based therapy due to partial loss of target antigen expression. PMID- 15548374 TI - Inositol hexaphosphate inhibits growth and induces G1 arrest and apoptotic death of androgen-dependent human prostate carcinoma LNCaP cells. AB - Prostate cancer (PCA) is the most common invasive malignancy and the second leading cause of cancer-related deaths in the US male population. One approach to control this malignancy is its preventive intervention by dietary agents. Inositol hexaphosphate (IP6), a dietary constituent, has shown promising efficacy against various cancers; however, limited studies have been performed with IP6 against PCA. Here, we investigated the growth-inhibitory effect and associated mechanisms of IP6 in androgen-dependent human prostate carcinoma LNCaP cells. IP6 treatment of cells resulted in a strong growth inhibition and an increase in G1 cell population. In mechanistic studies, IP6 resulted in an increase in cyclin dependent kinase inhibitors (CDKIs) Cip1/p21 and Kip1/p27 levels, together with a decrease in cyclin-dependent kinase (CDK) 4 and cyclin D1 protein levels. An increase in CDKI levels by IP6 also led to a concomitant increase in their interactions with CDK2 and CDK4, together with a strong decrease in the kinase activity of both CDKs. Downstream in CDKI-CDK-cyclin cascade, consistent with its inhibitory effect on CDK kinase activity, IP6 treatment of cells increased hypophosphorylated levels of retinoblastoma (Rb) with a decrease in Rb phosphorylation at serine 780, 807, and 811 sites, and caused a moderate to strong decrease in the levels of transcription factors E2F1, E2F4, and E2F5. In other studies, IP6 caused a dose- and a time-dependent apoptotic death of LNCaP cells, and a decrease in Bcl2 levels, causing a strong increase in Bax versus Bcl2 ratio, as well as an inhibition of constitutively active AKT phosphorylation. Taken together, these molecular alterations provide an insight into IP6-caused growth inhibition, G1 arrest, and apoptotic death of human prostate carcinoma LNCaP cells. Because early clinical PCA growth is an androgen dependent response, the results of the present study employing androgen-dependent LNCaP cells suggest that IP6 has promise and potential to be effective against PCA. PMID- 15548376 TI - Diagnosis of pancreatic cancer using serum proteomic profiling. AB - In the United States, mortality rates from pancreatic cancer (PCa) have not changed significantly over the past 50 years. This is due, in part, to the lack of early detection methods for this particularly aggressive form of cancer. The objective of this study was to use high-throughput protein profiling technology to identify biomarkers in the serum proteome for the early detection of resectable PCa. Using surface-enhanced laser desorption/ionization mass spectrometry, protein profiles were generated from sera of 49 PCa patients and 54 unaffected individuals after fractionation on an anion exchange resin. The samples were randomly divided into a training set (69 samples) and test set (34 samples), and two multivariate analysis procedures, classification and regression tree and logistic regression, were used to develop classification models from these spectral data that could distinguish PCa from control serum samples. In the test set, both models correctly classified all of the PCa patient serum samples (100% sensitivity). Using the decision tree algorithm, a specificity of 93.5% was obtained, whereas the logistic regression model produced a specificity of 100%. These results suggest that high-throughput proteomics profiling has the capacity to provide new biomarkers for the early detection and diagnosis of PCa. PMID- 15548375 TI - Mxi1-0, an alternatively transcribed Mxi1 isoform, is overexpressed in glioblastomas. AB - The c-Myc transcription factor regulates expression of genes related to cell growth, division, and apoptosis. Mxi1, a member of the Mad family, represses transcription of c-Myc-regulated genes by mediating chromatin condensation via histone deacetylase and the Sin3 corepressor. Mxi1 is a c-Myc antagonist and suppresses cell proliferation in vitro. Here, we describe the identification of Mxi1-0, a novel Mxi1 isoform that is alternatively transcribed from an upstream exon. Mxi1-0 and Mxi1 have different amino-terminal sequences, but share identical Max- and DNA-binding domains. Both isoforms are able to bind Max, to recognize E-box binding sites, and to interact with Sin3. Despite these similarities and in contrast to Mxi1, Mxi1-0 is predominantly localized to the cytoplasm and fails to repress c-Myc-dependent transcription. Although Mxi1-0 and Mxi1 are coexpressed in both human and mouse cells, the relative levels of Mxi1-0 are higher in primary glioblastoma tumors than in normal brain tissue. This variation in the levels of Mxi1-0 and Mxi1 suggests that Mxi1-0 may modulate the Myc-inhibitory activity of Mxi1. The identification of Mxi1-0 as an alternatively transcribed Mxi1 isoform has significant implications for the interpretation of previous Mxi1 studies, particularly those related to the phenotype of the mxi1 knockout mouse. PMID- 15548377 TI - Discovery and development of BVDU (brivudin) as a therapeutic for the treatment of herpes zoster. AB - This Commentary is dedicated to the memory of Dr. Jacques Gielen, the late Editor of Biochemical Pharmacology, whom I have known as both an author and reviewer for the Journal for about 25 years. This is, quite incidentally, about the time it took for bringing brivudin (BVDU) [(E)-5-(2-bromovinyl)-2'-deoxyuridine] from its original description as an antiviral agent to the market place (in a number of European countries, including Germany and Italy) for the treatment of herpes zoster in immunocompetent persons. BVDU is exquisitely active and selective against varicella-zoster virus (VZV) and herpes simplex virus type 1 (HSV-1). BVDU owes this high selectivity and activity profile to a specific phosphorylation by the virus-encoded thymidine kinase, followed by a potent interaction with the viral DNA polymerase. The (E)-5-(2-bromovinyl)-substituent can be considered as the hallmark for the activity of BVDU against VZV and HSV-1. Extensive clinical studies have indicated that BVDU as a single (oral) daily dose of 125 mg (for no more than 7 days) is effective in the treatment of herpes zoster, as regards both short-term (suppression of new lesion formation) and long term effects (prevention of post-herpetic neuralgia). In this sense, BVDU is as efficient and/or convenient, if not more so, than the other drugs (acyclovir, valaciclovir, famciclovir) that have been licensed for the treatment of herpes zoster. There is one caveat; however, BVDU should not be given to patients under 5-fluorouracil therapy, as the degradation product of BVDU, namely (E)-5-(2 bromovinyl)uracil (BVU), may potentiate the toxicity of 5-fluorouracil, due to inhibition of dihydropyrimidine dehydrogenase, the enzyme involved in the catabolism of 5-fluorouracil. PMID- 15548378 TI - Induction of alkaline phosphatase in the inflamed intestine: a novel pharmacological target for inflammatory bowel disease. AB - This study demonstrates the upregulation of alkaline phosphatase and the mechanisms involved in experimental colitis. All models of ileal and colonic inflammation examined, which were characterized by significant oxidative stress and neutrophil infiltration, resulted in an increase in alkaline phosphatase activity which was attributable to both epithelial cells and cells of the lamina propria, mainly leukocytes. The increase in alkaline phosphatase sensitivity to the inhibitors levamisole and homoarginine, together with changes in the apparent molecular size and in the sialization of the enzyme, indicated a change in the isoform expressed. An increase in tissue non-specific alkaline phosphatase expression was observed by Western blotting. Treatment with the bone/kidney alkaline phosphatase inhibitor levamisole or a monoclonal antibody resulted in significant protection from colonic inflammation. Taken together, these results indicate that the kidney isoform is a marker of intestinal inflammation and that it might even constitute a target for pharmacological intervention. PMID- 15548379 TI - Anti-inflammatory effect of caffeic acid methyl ester and its mode of action through the inhibition of prostaglandin E2, nitric oxide and tumor necrosis factor-alpha production. AB - The anti-inflammatory effects of caffeic acid (CA), caffeic acid methyl ester (CM) and di-O-acetylcaffeic acid (DAC) were investigated in rats using the carrageenin-induced edema model and the antinociceptive effects of these compounds were also assessed in mice by means of the acetic acid-induced abdominal constriction test and hot plate test. CM (10mg/kg, p.o.) showed the most potent anti-inflammatory and antinociceptive effects in these animal models. To investigate the mechanism of the anti-inflammatory action, we examined the effects of these compounds on the lipopolysaccharide (LPS)-induced NO and PGE2 responses in the murine macrophage cell line, RAW 264.7. Our data indicate that CM is the most potent inhibitor of NO and PGE2 production and it also significantly decreased tumor necrosis factor-alpha (TNF-alpha) release. Consistent with these observations, the protein and mRNA expression levels of iNOS and COX-2 were found to be inhibited by CM in a dose-dependent manner. Furthermore, CM inhibited the nuclear factor-kappaB (NF-kappaB) activation induced by LPS, which was associated with the prevention of the degradation of the inhibitor kappaB, and subsequently with decreased p65 protein levels in the nucleus. Taken together, our data indicate that the anti-inflammatory properties of CM might result from the inhibition of iNOS, COX-2 and TNF-alpha expression through the down-regulation of NF-kappaB binding activity. PMID- 15548380 TI - Polymerization of the triphosphates of AraC, 2',2'-difluorodeoxycytidine (dFdC) and OSI-7836 (T-araC) by human DNA polymerase alpha and DNA primase. AB - OSI-7836 (4'-thio-araC, T-araC) is a nucleoside analogue that shows efficacy against solid tumor xenograft models. We examined how the triphosphates of OSI 7836 (T-araCTP), cytarabine (araCTP), and gemcitabine (dFdCTP) affected the initiation of new DNA strands by the pol alpha primase complex. Whereas dFdCTP very weakly inhibited primase, both T-araCTP and araCTP potently inhibited this enzyme. Primase polymerized T-araCTP and araCTP more readily than its natural substrate, CTP, and incorporation resulted in strong chain termination. dFdCTP, araCTP, and T-araCTP inhibited pol alpha competitively with respect to dCTP. When exogenously added primentemplates were used, pol alpha incorporated all three analogues into DNA, and incorporation caused either weak chain termination (dFdCTP), strong termination (araCTP), or extremely strong termination (T-araC). Furthermore, pol alpha polymerized T-araCTP only nine-fold less well than dCTP, whereas it polymerized araCTP and dFdCTP 24- and 83-fold less well, respectively. The presence of these three analogues in the template strand resulted in significant pausing by pol alpha, although the site and severity of pausing varied between the analogues. During the elongation of primase-synthesized primers, a reaction that is thought to mimic the normal sequence of events during the initiation of new DNA strands, pol alpha polymerized all three compounds. However, incorporation of araCTP and dFdCTP resulted in minimal chain termination, while incorporation of T-araCTP still caused extremely strong termination. The implications of these results with respect to how these compounds affect cells are discussed. PMID- 15548381 TI - A PXR reporter gene assay in a stable cell culture system: CYP3A4 and CYP2B6 induction by pesticides. AB - A stable hepatoma cell line expressing the human pregnane X receptor (hPXR) and the cytochrome P4503A4 (CYP3A4) distal and proximal promoters plus the luciferase reporter gene was developed to assess the ability of several xenobiotic agents to induce CYP3A4 and CYP2B6. After selection for neomycin resistance, one clone, displaying high luciferase activity in response to rifampicin (RIF), was isolated and the stable expression of hPXR was confirmed by reverse transcription polymerase chain reaction (RT-PCR). Dose-response curves were generated by treating these cells with increasing concentrations of RIF, phenobarbital (PB), clotrimazole (CLOT) or 5beta-pregnane-3,20-dione (5beta-PREGN). The effective concentrations for half maximal response (EC50) were determined for each of these compounds. RIF was the most effective compound, with maximal luciferase activity induced at 10 microM. The agonist activities of PXR-specific inducers measured using our stable model were consistent with those measured in transient transfectants. The abilities of organochlorine (OC), organophosphate (OP) and pyrethroid pesticides (PY) to activate hPXR were also assessed and found to be consistent with the abilities of these compounds to induce CYP3A4 and CYP2B6 in primary culture of human hepatocytes. These results suggest that CYP3A4 and CYP2B6 regulation through PXR activation by persistent pesticides may have an impact on the metabolism of xenobiotic agents and endogenous steroid hormones. Our model provides a useful tool for studying hPXR activation and for identifying agents capable of inducing CYP3A4 and CYP2B6. PMID- 15548382 TI - Inhibition of angiogenesis in vivo and growth of Kaposi's sarcoma xenograft tumors by the anti-malarial artesunate. AB - Artesunate (ART) is a semi-synthetic derivative of the sesquiterpene artemisinin used for the second line therapy of malaria infections with Plasmodium falciparum. ART also inhibits growth of many transformed cell lines. In the present investigation, we show that ART inhibited the growth of normal human umbilical endothelial cells and of KS-IMM cells that we have established from a Kaposi's sarcoma lesion obtained from a renal transplant patient. The growth inhibitory activity correlated with the induction of apoptosis in KS-IMM cells. Apoptosis was not observed in normal endothelial cells, which, however, showed drastically increased cell doubling times upon ART treatment. ART strongly reduced angiogenesis in vivo in terms of vascularization of Matrigel plugs injected subcutaneously into syngenic mice. We conclude that ART represents a promising candidate drug for the treatment of the highly angiogenic Kaposi's sarcoma. As a low-cost drug, it might be of particular interest for areas of Kaposi's sarcoma endemics. ART could be useful for the prevention of tumor angiogenesis. PMID- 15548383 TI - Effects and regulation of osteopontin in rat hepatic stellate cells. AB - Using a cDNA microarray, we identified osteopontin (OPN) as one of the genes upregulated in cultured activated hepatic stellate cells (HSCs). Northern and western blot analyses showed that OPN was increasingly expressed during the progressive activation of cultured rat HSCs, and a significant increase in OPN was observed in carbon tetrachloride-induced rat liver fibrosis. In biliary atresia, OPN protein was predominantly expressed in Kupffer cells and HSCs in the necrotic areas. Incubation of HSCs with recombinant OPN-induced significant proliferative and migratory effects, and induced matrix metalloproteinase 2 production and activation. Moreover, OPN increased type I collagen production and type II transforming growth factor-beta receptor mRNA and protein. In conclusion, this study shows that OPN is expressed in activated HSCs and suggests that the upregulation of OPN might be a central pathway of HSC activation. PMID- 15548384 TI - Design of vesicles of 1,2-di-O-acyl-3-O-(beta-D-sulfoquinovosyl)-glyceride bearing two stearic acids (beta-SQDG-C18), a novel immunosuppressive drug. AB - The immunosuppressive effects of synthetic sulfo-glycolipids in the class of sulfoquinovosyl-diacylglycerols (SQDG), including stereoisomers, were interesting in development of a promising clinical drug. Especially, 1,2-di-O-stearoyl-3-O-(6 deoxy-6-sulfo-beta-D-glucopyranosyl)-sn-glycerol (beta-SQDG-C18) was thought to be a valuable candidate because of the preliminary observations of its high inhibitory activities in spite of low toxicities. The problem of using this material is to find an applicable way avoiding its low solubility in water. The vesicle formation of beta-SQDG-C18 is advantageous to i.v. administration in its chemico-structural character. With preparation in water, beta-SQDG-C18 was hard to form vesicles, because its hydrophilicity was strong. We examined the suitable parameter of the vesicle forming condition. It was possible to take a balance between the hydrophilicity and the hydrophobicity of the beta-SQDG-C18 molecule to be optimized to form vesicles in 150 mM PBS. In addition, we demonstrated the strong immunosuppressive activity of beta-SQDG-C18 vesicles. This is the first report of the preparation method of beta-SQDG-C18 vesicles, which should facilitate in vitro and in vivo application. PMID- 15548385 TI - Involvement of both endoplasmic reticulum and mitochondria in photokilling of nasopharyngeal carcinoma cells by the photosensitizer Zn-BC-AM. AB - Photodynamic therapy (PDT) is recently developed as an effective treatment for malignant disease. In PDT, the photosensitizer eradicates tumour by induction of apoptosis. In this study, we investigated the mechanistic actions of a recently developed second generation photosensitizer, Zn-BC-AM, on nasopharyngeal carcinoma (NPC) cells. Zn-BC-AM was found to localize in the mitochondria, endoplasmic reticulum (ER), and golgi body. Photoactivation of Zn-BC-AM loaded NPC cells resulted in a rapid collapse of mitochondrial membrane potential (Deltapsim) (15 min), followed by the release of cytochrome c (1 h), and activation of caspases-9 and -3 (4 h). Expression of ER chaperones Bip/Grp78 and Grp94, and ER resident lectin-like chaperone calnexin (CNX) was also enhanced in PDT-stressed NPC cells. Caspase-12, an important caspase involved in ER stress induced apoptosis, was also activated. Inhibition of Ca2+ uptake into mitochondria by ruthenium red (RR) or loading the cells with EGTA-AM, an agent that buffers intracellular Ca2+ released from ER, resulted in a significant reduction of Zn-BC-AM PDT-induced cell death. These observations suggest that both ER and mitochondria are the subcellular targets of Zn-BC-AM. Effective activation of ER- and mitochondria-mediated apoptotic pathways is responsible for Zn-BC-AM PDT-induced NPC cell death. PMID- 15548386 TI - Transcriptional regulation of the mouse deoxycytidine kinase: identification and functional analysis of nuclear protein binding sites at the proximal promoter. AB - Deoxycytidine kinase (EC 2.7.1.74, dCK) catalyzes the phosphorylation of deoxynucleosides and several nucleoside analogues that are important in antiviral and cancer chemotherapy. The enzyme is predominantly expressed in lymphoid tissue by as yet poorly defined mechanisms. In this work, we have studied the mouse dCK regulatory region to understand the molecular details of the tissue specific expression of the enzyme. DNase I footprinting and electrophoretic mobility shift assays using nuclear extracts from mouse lymphocytes (EL-4, T cells; J558, B cells) and non-lymphoid cells (L929, fibroblasts) demonstrated the existence of at least six cis-acting elements (FP-1-FP-6) within the proximal promoter region. Functional analysis revealed that all the elements necessary to promote high level transcription of the mdCK gene are located downstream the transcription start site. 5'-Deletion and site-directed mutagenesis assays demonstrated the importance of four GC-rich regions, which bind Sp-1 and Sp-3 transcription factors. In addition, we identified a site (FP-3) located at the -282 to -310 nucleotide region of the promoter, which binds NF-1, only in B cells. Analysis of point mutations introduced at the different regions revealed functional differences in their role in mdCK transcription in the cell lines used. PMID- 15548387 TI - In vitro and ex vivo evidence for modulation of P-glycoprotein activity by progestins. AB - The well known gender-related differences in drug action may partly be explained by changes in activity and expression of drug metabolising enzymes, but also by modulation of active drug transport systems (e.g. P-glycoprotein, Pgp) by sexual steroids, which is yet not well investigated. Because many women are using hormones (e.g. as oral contraceptives) we investigated the influence of different synthetic progestins on Pgp activity. Pgp inhibition of progesterone, medroxyprogesterone, chlormadinone, cyproterone, levonorgestrel, norethisterone, desogestrel, and norgestimate was measured in vitro in two Pgp over-expressing cell lines (L-MDR1, P388/dx cells) and the corresponding parental cell lines by means of calcein assay, and ex vivo in human peripheral blood mononuclear cells (PBMCs) by rhodamine123 efflux. For most progestins tested, concentrations needed to double baseline fluorescence (f2) in L-MDR1 cells were similar to that of the potent Pgp inhibitor quinidine, whereas levonorgestrel and norethisterone did not reach f2. The results in P388/dx cells essentially confirmed our findings in L MDR1 cells. Additionally, Pgp inhibitory activity of all progestins tested was also shown ex vivo in PBMCs. The potent Pgp inhibition by several synthetic progestins in vitro and ex vivo suggests that such an interaction might be clinically relevant despite generally low plasma concentrations of progestins. The results may be of particular importance for Pgp substrates, such as protease inhibitors and chemotherapeutic agents, for which intracellular concentrations are critical. PMID- 15548388 TI - Human CYP4F12 genetic polymorphism: identification and functional characterization of seven variant allozymes. AB - The human cytochrome CYP4F12 has been shown to be metabolically active toward inflammatory mediators and exogenous compounds such as antihistaminic drugs. We recently identified a genetic polymorphism within the promoter region, associated with a decreased level of enzyme expression. In the present study, we report the further identification of single nucleotide polymorphisms in the coding sequence of the CYP4F12 gene. A polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis of DNA samples from 53 unrelated French Caucasians, allowed the identification of ten mutations, comprising seven missense mutations, 31C>T (Leu11Phe), 38C>T (Pro13Leu), 47C>T (Met16Thr), 4759G>A (Asp76Asn), 4801G>A (Val90Leu), 8896C>T (Arg188Cys) and 23545G>A (Gly522Ser). Their functional impact toward ebastine hydroxylation was evaluated using heterologous expression in Saccharomyces cerevisiae cells of site-directed mutated cDNA variants. Five out seven variants did not exhibit any significant difference in CYP4F12 catalytic activity, whereas two variants, Val90Ile and Arg188Cys, displayed significant changes in their Michaelis-Menten (Km, Vm) parameters. These data on CYP4F12 genetic polymorphism provide tools for further studies of association with pathological processes involving an inflammatory component and with variations in anti-histaminic drug response. PMID- 15548389 TI - The immunosuppressant drug FK506 prevents Fas-induced apoptosis in human hepatocytes. AB - FK506 is a potent immunosuppressive drug used for the prevention of graft rejection in organ transplantation. Experimental and clinical studies have shown correlations between apoptosis and graft rejection, and apoptosis also plays a role in cell death after ischemia-reperfusion injury in the rat liver. Fas mediated apoptosis is very likely involved in allograft rejection and experimental evidence has shown a decrease of FasR expression in mouse hepatocytes produced by the drugs. On the basis of these findings we have investigated the protective effect of FK506 in comparison with cyclosporine A (CsA) on Fas-induced apoptosis, by analysing the activation of downstream effector caspases in human hepatocytes. Apoptosis was induced by treatment with agonistic antibodies against FasR, which resulted in a significant activation of caspase-3 after 12 h. Prevention of the downstream activation of the caspase cascade and apoptosis was observed when hepatocytes were pre-treated for 3 h with immunosuppressant drugs. A significant reduction (ca. 30-40%) of caspase-3 activation by 5 microM FK506 and CsA was observed. Along with less activation of caspase-3 a decrease of apoptotic DNA fragmentation was found. In addition, FK506 significantly reduced not only caspase-8 but also caspase-9 activation, to a similar extent as CsA, thus suggesting a protective effect at the mitochondrial level of this drug, as has already been reported for CsA. These effects of FK506 help to explain its strong anti-rejection properties and suggest promising benefits of pharmacological preconditioning on ischemia-reperfusion injury following liver transplantation. PMID- 15548390 TI - Enhancement of paclitaxel-induced microtubule stabilization, mitotic arrest, and apoptosis by the microtubule-targeting agent EM012. AB - EM012, a semisynthetic phthalideisoquinoline alkaloid, has been recently found to target microtubules and possess anti-cancer activity. In this study, we evaluated the effects of EM012 in combination with the classic microtubule-targeting agent paclitaxel. Our results demonstrated that EM012 enhanced the anti-proliferative activity of nanomolar concentrations of paclitaxel in human breast cancer (MCF7), prostate cancer (DU145), and non-small-cell lung cancer (A549) cells. Further studies revealed that EM012 increased the ability of nanomolar concentrations of paclitaxel to induce mitotic arrest and apoptosis, without affecting microtubule polymerization. In contrast, when micromolar concentrations of paclitaxel were used, EM012 promoted paclitaxel-induced microtubule polymerization both in vitro and in cultured cells. Nevertheless, EM012 enhanced the ability of nanomolar concentrations of paclitaxel to stabilize microtubules, as indicated by increased tubulin acetylation. Our results therefore suggest a therapeutic potential of EM012/paclitaxel combination in the management of human cancer and provide mechanistic insights into the combined effects of these two microtubule-targeting agents. PMID- 15548391 TI - The interactions between the N-terminal and C-terminal domains of the human UDP glucuronosyltransferases are partly isoform-specific, and may involve both monomers. AB - The pathological mutation Y486D was previously shown to reduce the activities of the UDP-glucuronosyltransferases (UGTs) 1A1 and 1A6 by about 88% and 99%, respectively. Surprisingly, the corresponding mutation in UGT1A9 (Y483D) doubled the Vmax of scopoletin glucuronidation, whereas the entacapone glucuronidation rate was decreased by about 50%. Due to the primary structure identity of the C terminal half of all the human UGTs of the 1A subfamily, the sharp differences between them in the effect of a mutation deep inside the C-terminal half suggested that there are isoform-specific interactions between the variable N- and the conserved C-terminal halves. In dimeric enzymes, like the UGTs, such interactions might either occur within the same polypeptide, or between opposite monomers. The latter implies functional monomer-monomer interactions, and this was investigated using hetero-dimeric UGTs. Insect cells were co-infected with mixtures containing different combinations of recombinant baculoviruses encoding either UGT1A4 or 1A9Sol. The UGT1A4 was selected because it glucuronidates neither entacapone nor scopoletin at significant rates. The active enzyme in these hetero-dimers was 1A9Sol, a truncation mutant of UGT1A9 that exhibited a very low ratio of entacapone to scopoletin glucuronidation rates. Interestingly, the ratio of entacapone to scopoletin glucuronidation rates in the co-infected cells was dependent on, and markedly increased with, the probability that 1A9Sol forms hetero-dimers with UGT1A4. In addition, the apparent Km for entacapone in the hetero-dimers was much lower than in 1A9Sol, and resembled the corresponding value in full-length UGT1A9. The results, thus, revealed important monomer monomer interactions within the UGTs. PMID- 15548392 TI - Ultrasonographic evaluation of lower uterine segment thickness in patients of previous cesarean section. AB - OBJECTIVE: To evaluate by ultrasonography, the lower uterine segment thickness of women with a previous cesarean delivery and determine a critical thickness above which safe vaginal delivery is predictable. METHODS: A prospective observational study of 71 antenatal women with previous cesarean delivery and 50 controls was carried out. Transabdominal and transvaginal ultrasonography were used in both groups to evaluate lower uterine segment thickness. The obstetric outcome in patients with successful vaginal birth and intraoperative findings in women undergoing cesarean delivery were correlated with lower segment thickness. RESULTS: The overall vaginal birth after cesarean section (VBAC) was 46.5% and VBAC success rate was 63.5%, the incidence of dehiscence was 2.82%, and there were no uterine ruptures. There was a 96% correlation between transabdominal ultrasonography with magnification and transvaginal ultrasonography. The critical cutoff value for safe lower segment thickness, derived from the receiver operator characteristic curve, was 2.5 mm. CONCLUSION: Ultrasonographic evaluation permits better assessment of the risk of scar complication intrapartum, and could allow for safer management of delivery. PMID- 15548393 TI - Risk factors and obstetric complications associated with macrosomia. AB - OBJECTIVE: Macrosomia is associated with adverse maternal outcomes. The objective of this study was to characterize the epidemiology of macrosomia and related maternal complications. METHOD: Live births (146,526) were identified between 1995 and 1999 in the Kaiser Permanente Medical Care Program's Northern California Region (KPMCP NCR) database. Bivariate and multivariate analyses were performed for risk factors and complications associated with macrosomia (birth weight >4500 g). RESULT: Male infant sex, multiparity, maternal age 30-40, white race, diabetes, and gestational age >41 weeks were associated with macrosomia (p<0.001). In bivariate and multivariate analyses, macrosomia was associated with higher rates of cesarean birth, chorioamnionitis, shoulder dystocia, fourth degree perineal lacerations, postpartum hemorrhage, and prolonged hospital stay (p<0.01). CONCLUSION: Macrosomia was associated with adverse maternal outcomes in this cohort. More research is needed to determine how to prevent complications related to excessive birth weight. PMID- 15548394 TI - Multiple cesarean section morbidity. AB - OBJECTIVE: To quantify maternal risk associated with multiple cesarean sections (CSs) and determine whether the third CS defines a threshold for increased morbidity. METHODS: From January 1997 to January 2002, the clinical records of 3191 women who were delivered by CS at our referral maternity center were examined for selected indicators of maternal morbidity. The women were assigned to groups based on number of CSs and the frequency of each indicator was determined. A composite score for each indicator among women grouped by number of consecutive CSs was then derived to compare risk between groups and against the third CS. RESULTS: By all indicators studied, morbidity increased with successive CSs before and through the third CS. However, compared with the third, the risk of major morbidity was significantly increased with the fifth, and much worse at the sixth CS for placenta previa (odds ratio [OR]=3.8, 95% confidence interval [CI]=1.9-7.4), placenta accreta (OR=6.1, 95% CI=2.0-18.4) and hysterectomy (OR=5.9, 95% CI=1.5-24.4). But the third and fourth CSs had the same risk of major morbidity for placenta previa (OR=1.4, 95% CI=0.8-2.2), placenta accreta (OR=1.0, 95% CI=0.3-2.9) and hysterectomy (OR=0.3, 95% CI=0.0-2.7). CONCLUSIONS: The third CS does not define a threshold for increased risk to the mother. Instead, overall morbidity rises continually with each successive CS. However, specifically for major morbidity from the triad of placenta previa, placenta accreta and hysterectomy during CS, the fourth CS carries the same risk as the third. PMID- 15548395 TI - Median and ulnar nerve conduction in pregnancy. AB - OBJECTIVE: A prospective, observational study assessed median and ulnar nerve conduction during pregnancy, to identify the optimum test for differentiating physiological effects of pregnancy from pathological carpal tunnel syndrome (CTS). METHODS: Pregnant women (n=18) and age- and parity-matched non-pregnant controls (n=13) were recruited. Symptomatic and neurophysiological evaluations were performed. Median and ulnar nerve latencies and intrapalmar latency (difference between median and ulnar nerve latencies) were computed. RESULTS: Median nerve distal latency is more prolonged in pregnant women compared with non pregnant controls. Median nerve latency is more prolonged in pregnant women with symptoms of CTS than in asymptomatic women. The difference between median and ulnar nerve conduction (normal <0.2 ms) discriminates well between symptomatic and asymptomatic pregnant women. CONCLUSIONS: Intrapalmar latency is proposed as a useful diagnostic test for classification of carpal tunnel syndrome during pregnancy. PMID- 15548396 TI - Metformin therapy in women with polycystic ovary syndrome. AB - OBJECTIVE: To determine the clinical, biochemical, hormonal, and ultrasonographic effects of 6 months of metformin therapy in women with polycystic ovary syndrome (PCOS) and compare with pretherapy parameters. METHOD: 50 Indian women with PCOS, 25 unmarried and 25 married, infertile women, were enrolled in this prospective clinical study. After a baseline workup, including body mass index (BMI), waist hip ratio (WHR), Ferriman Gallwey hirsutism scoring, menstrual pattern, levels of fasting insulin, lipids, oral glucose tolerance test (OGTT), serum gonadotropins, estradiol (E2), testosterone, androstenedione, sex hormone binding globulin (SHBG), and dehydroepiandrosterone sulphate (DHEAS), patients were given 1000 gm of metformin for 6 months and then reevaluated. RESULT: In 41 of 50 women who completed treatment, significant improvement in BMI, WHR, menstrual cyclicity (80.5%), ovulation rate (66%), and pregnancy rate (28%) was noted. Statistically significant decrease in lutenising hormone (LH) and LH/FSH ratio with an increase in follicle stimulating hormone (FSH) levels were seen. Levels of high-density lipoprotein (HDL) cholesterol (Chol) increased along with a decrease in total cholesterol. Improvement was noted in ovarian volume, stromal thickness, and number of follicles. There was no change in hirsutism, acne, levels of other sex steroid hormones, and lipids. CONCLUSION: A 6-month course of metformin therapy may improve menstrual cyclicity and fertility in women with PCOS. PMID- 15548397 TI - Falsely reassuring short-term variation associated with severe fetal acidemia in a near-term pregnancy. PMID- 15548398 TI - Genital flora in pregnancy and its association with group B streptococcal colonization. PMID- 15548399 TI - Human T cell leukemia virus-1 in pregnancy. PMID- 15548400 TI - Uterine artery embolization for the treatment of symptomatic fibroids. PMID- 15548401 TI - Extra-pelvic endometriosis presenting as a vulvar mass in a teenage girl. PMID- 15548402 TI - Identification of c-kit-positive cells in the uterus. PMID- 15548403 TI - Immunohistochemical analysis of IGF-1 and IGF-2 receptors in ovaries of patients with polycystic ovary syndrome. PMID- 15548404 TI - Ovarian cyst formation and in vitro fertilization outcome. PMID- 15548405 TI - Gender and sexual health in clinical practice in Lebanon. AB - This study examines the readiness of obstetricians/gynecologists (Ob/Gyns) in Lebanon to provide sexual consultation, their degree of comfort when discussing issues of sexual health, and their attitudes regarding assessment, treatment, and referral. Data on these concepts were collected through face-to-face interviews with 286 randomly selected Ob/Gyns. Most Ob/Gyns reported feeling comfortable discussing sexuality during consultations, which they attributed much more to professional experience than to training. Most Ob/Gyns reported giving proper time for management of sexual health issues and follow-up, as these issues are brought up frequently by their women clients. However, results suggest that only one-third (31%) of Lebanese Ob/Gyns nearly always take the initiative in asking patients about their sexual health. Moreover, almost 45% of participating Ob/Gyns did not recognize a strong relationship between reproductive health and sexual functioning. Gender was not found to be an important predictor for any of the indicators measured in the present study. Ob/Gyns in Lebanon are significant consultants on various sexual issues, and they need better postgraduate training, continuing medical education, and access to medical congress resources on the topic of sexuality and its relationship to reproductive health. PMID- 15548406 TI - Conducting international collaborative research in developing nations. AB - International research partnerships bring together some of the best and the brightest in an effort to tackle global health problems. Such collaborations also pose complex challenges, such as maintaining ethical principles in the conduct of research in developing nations. In implementing a randomized clinical trial to reduce postpartum hemorrhage (PPH) during childbirth in rural India, U.S. and Indian collaborators addressed three such issues: the appropriateness of an ethical randomized controlled trial in the developing world, the inclusion of a placebo arm, and the relevance of informed consent in a semiliterate rural population. PMID- 15548407 TI - Management of breech presentation in areas with high prevalence of HIV infection. AB - OBJECTIVE: To provide recommendations for the management of breech presentation in areas of high prevalence of human immunodeficiency virus (HIV) infection. METHOD: Review of relevant literature. RESULTS: Studies show that elective cesarean section (CS) is safer than vaginal delivery for breech presentation, external cephalic version (ECV) at term increases the chance of vaginal cephalic delivery. Although there are no studies of the risk of mother-to-child transmission of HIV from ECV, indirect evidence suggests that any increased risk is likely to be very small. RECOMMENDATIONS: Where CS is available and safe, HIV positive women, or women who might be at risk of HIV, with a fetus at term with breech presentation, should be offered elective CS to reduce the risks of both vaginal breech delivery and mother-to-child HIV infection. HIV-negative women can be offered ECV at term to try to avoid CS. Where women do not have access to a safe CS, or prefer vaginal delivery, the benefit for both mother and child of attempting ECV at term is likely to outweigh the theoretical, very small, risk of facilitating HIV transmission. PMID- 15548408 TI - Observations on reproductive health programs in the Baltic States. AB - Public attention in Sweden has been drawn to three neighboring states that recently joined the European Union: Estonia, Latvia, and Lithuania. At this historic moment, it seems instructive to look at how the rapidly reformed health sectors of these ex-Soviet republics are responding to the vision of reproductive health articulated in Cairo 10 years ago. Reproductive health and rights have improved in these states in spite of recent reforms often acting to oppose improvement. Reforms such as the introduction of family medicine need continued adjustment, especially regarding antenatal care. One special challenge is the retention of essential mid-level providers, such as midwives, as the mode of HIV transmission becomes increasingly sexual. PMID- 15548409 TI - A consensus regimen for early abortion with misoprostol. PMID- 15548410 TI - Editor's comment. PMID- 15548411 TI - Strategies to prevent eclampsia in a developing country: I. Reorganization of maternity services. AB - OBJECTIVE: To determine whether changes in primary and secondary care service delivery could prevent antenatal eclampsia. METHOD: One intervention (St. Catherine) and two control (St. Ann, Manchester) parishes were chosen. The health system in St. Catherine was restructured. Primary antenatal clinics had clear instructions for referring patients to a high-risk antenatal clinic or to hospital. Guidelines were provided to high-risk clinics and the antenatal ward for appropriate treatment of hypertension and preeclampsia when induction of labor should occur. Antenatal eclampsia incidence was monitored before and during the intervention and compared with control parishes (no intervention). Each eclampsia case was investigated to identify inadequacies in the system. RESULTS: The process resulted in better identification of women at risk. Antenatal eclampsia incidence dropped dramatically as care improved. Compared with control areas, by completion of the study, the rate was significantly lower than at the start: OR 0.19 (95% CI: 0.13-0.27; p<0.001 trend). Antenatal admissions for hypertensive disorders declined significantly, and the number of bed days halved. CONCLUSION: Reorganization of maternal care can have major public health benefits and cost savings; however, women need to be alerted to recognise and act upon signs of impending eclampsia. PMID- 15548412 TI - Strategies to prevent eclampsia in a developing country: II. Use of a maternal pictorial card. AB - OBJECTIVE: To assess the efficacy and acceptability of a patient-held pictorial card aimed at raising awareness and appropriate health seeking behavior in response to prodromal symptoms of imminent eclampsia. METHOD: Pictorial cards (and posters) were issued to antenatal clinics and used to focus instruction and advice to pregnant women. Mothers were surveyed before and after the cards were introduced to assess maternal likelihood of seeking care if edema was seen, and of attending hospital if so advised. We monitored the eclampsia rate. Health workers were interviewed 6 months after cards and posters were issued to determine the acceptability of using the cards as part of routine antenatal care. RESULTS: The card was seen as widely acceptable by health professionals, and increased their own awareness of the prodromal symptoms of eclampsia and their discussion of these symptoms with antenatal mothers. Mothers' awareness and response to symptoms improved significantly and there was a marked drop in eclampsia incidence. Suggested improvements to the card were made by mothers and health workers. CONCLUSION: The cost of providing a card for every pregnant mother is likely to be offset by health service delivery savings. PMID- 15548413 TI - What do doctors think their caseload should be to maintain their skills for delivery care? AB - OBJECTIVES: To seek the opinions of doctors on what they thought the minimum, maximum and optimum annual caseload should be for the maintenance of skills and competence in a variety of obstetric procedures. METHODS: An expert panel of respondents from Africa and Asia was asked to give their opinions on what they believed the minimum, maximum and optimum caseload should be for 11 obstetric procedures via a series of structured questionnaires (Delphi exercise). In subsequent questionnaires, participants were asked if they wished to reconsider their opinions in light of the group response. RESULTS: The median values of responses given for the minimum, maximum and optimum caseloads for the 11 obstetric procedures did not change substantially over time, though greater consensus was developed as indicated by reductions in the size of inter-quartile ranges in later rounds. CONCLUSIONS: We encountered several problems associated with using the Delphi technique in this context, which throws doubt on the validity and usefulness of our results. Caseload is just one of many factors, as indicated by our expert panel that need to be considered when planning the delivery of obstetric services in remote areas. High quality training, continued medical education, appropriate quality assurance procedures, and provision of a supportive enabling environment are also important requirements. In addition, the views of clinicians need to be balanced against more objective evidence of quality of care and patient outcome in relation to procedural volume. Such evidence is lacking in the field of obstetrics and requires further investigation. PMID- 15548414 TI - Therapeutic strategies for Duchenne and Becker dystrophies. AB - Duchenne muscular dystrophy (DMD), a severe X-linked genetic disease affecting one in 3500 boys, is the most common myopathy in children. DMD is due to a lack of dystrophin, a submembrane protein of the cytoskeleton, which leads to the progressive degeneration of skeletal, cardiac, and smooth muscle tissue. A milder form of the disease, Becker muscular dystrophy (BMD), is characterized by the presence of a semifunctional truncated dystrophin, or reduced levels of full length dystrophin. DMD is the focus of three different supportive or therapeutic approaches: gene therapy, cell therapy, and drug therapy. Here we consider these approaches in terms of three potential goals: improvement of dystrophic phenotype, expression of dystrophin, and overexpression of utrophin. Utrophin exhibits 80% homology with dystrophin and is able to perform similar functions. Pharmacological strategies designed to overexpress utrophin appear promising and may circumvent many obstacles to gene and cell-based therapies. PMID- 15548415 TI - Cellular distribution and functions of P2 receptor subtypes in different systems. AB - This review is aimed at providing readers with a comprehensive reference article about the distribution and function of P2 receptors in all the organs, tissues, and cells in the body. Each section provides an account of the early history of purinergic signaling in the organ?cell up to 1994, then summarizes subsequent evidence for the presence of P2X and P2Y receptor subtype mRNA and proteins as well as functional data, all fully referenced. A section is included describing the plasticity of expression of P2 receptors during development and aging as well as in various pathophysiological conditions. Finally, there is some discussion of possible future developments in the purinergic signaling field. PMID- 15548416 TI - Neuropeptides in hypothalamic neuronal disorders. AB - A few examples of hypothalamic, peptidergic disorders leading to clinical signs and symptoms are presented in this review. Increased activity of corticotropin releasing hormone (CRH) neurons in the paraventricular nucleus (PVN) and decreased activity of the vasopressin neurons in the biological clock and of the thyroxine-releasing hormone (TRH) neurons in the PVN contribute to the signs and symptoms of depression. In men, the central nucleus of the bed nucleus of the stria terminalis (BSTc) is about twice as large and contains twice as many somatostatin neurons as in women. In transsexuals this sex difference is reversed, pointing to a role of this structure in gender. Luteinizing hormone releasing hormone (LHRH) neurons are formed in the fetal olfactory placade and migrate along the terminal nerve fibers into the hypothalamus. In Kallmann's syndrome the migration process of the LHRH (gonadotropin-releasing hormone) neurons is aborted, which explains the joint occurrence of hypogonadotropic hypogonadism and anosmia in this syndrome. In postmenopausal women, the neurons of the infundibular nucleus hypertrophy and become hyperactive because of the disappearance of the estrogen feedback and contain hyperactive peptidergic neurons. Climacteric flushes may be caused by hyperactivity of the neurokinin-B or LHRH neurons in this nucleus. The hypocretin (orexin) neurons in the perifornical area are involved in sleep. In narcolepsy with cataplexy, a loss of these neurons, probably due to an autoimmune process, is found. Obese subjects with a mutation in the gene that encodes for leptin, the preproghrelin gene, or the alpha-melanocyte-stimulating hormone (alpha-MSH) gene have been described. Decreased numbers and activity of the oxytocin neurons in the PVN may be responsible for the absence of satiety in Prader-Willi syndrome. Moreover, a glucocorticoid receptor polymorphism is associated with obesitas and dysregulation of the hypothalamus-pituitary-adrenal axis. In contrast, two single nucleotide polymorphisms (SNPs) of the AGRP gene have been associated with anorexia nervosa. PMID- 15548417 TI - Multiple parallelisms in animal cytokinesis. AB - The process of cytokinesis in animal cells is usually presented as a relatively simple picture: A cleavage plane is first positioned in the equatorial region by the astral microtubules of the anaphase mitotic apparatus, and a contractile ring made up of parallel filaments of actin and myosin II is formed and encircles the cortex at the division site. Active sliding between the two filament systems constricts the perimeter of the cortex, leading to separation of two daughter cells. However, recent studies in both animal cells and lower eukaryotic model organisms have demonstrated that cytokinesis is actually far more complex. It is now obvious that the three key processes of cytokinesis, cleavage plane determination, equatorial furrowing, and scission, are driven by different mechanisms in different types of cells. In some cases, moreover, multiple pathways appear to have redundant functions in a single cell type. In this review, we present a novel hypothesis that incorporates recent observations on the activities of mitotic microtubules and the biochemistry of Rho-type GTPase proteins and postulates that two different sets of microtubules are responsible for the two known mechanisms of cleavage plane determination and also for two distinct mechanisms of equatorial furrowing. PMID- 15548418 TI - Biosynthesis and alternate targeting of the lysosomal cysteine protease cathepsin L. AB - Upregulation of cathepsin L expression, whether during development or cell transformation, or mediated by ectopic expression from a plasmid, alters the targeting of the protease and thus its physiological function. Upregulated procathepsin L is targeted to small dense core vesicles and to the dense cores of multivesicular bodies, as well as to lysosomes and to the plasma membrane for selective secretion. The multivesicular vesicles resemble secretory lysosomes characterized in specialized cell types in that they are endosomes that stably store an upregulated protein and they possess the tetraspanin CD63. Morphologically the multivesicular endosomes also resemble late endosomes, but they store procathepsin L, not the active protease, and they are not the major site for LAMP-1 accumulation. Distinction between the lysosomal proenzyme and active protease thus identifies two populations of multivesicular endosomes in fibroblasts, one a storage compartment and one an enzymatically active compartment. A distinctive targeting pathway using aggregation is utilized to enrich the storage endosomes with a particular lysosomal protease that can potentially activate and be secreted. PMID- 15548419 TI - Microtubule-associated proteins and their essential roles during mitosis. AB - Microtubules play essential roles during mitosis, including chromosome capture, congression, and segregation. In addition, microtubules are also required for successful cytokinesis. At the heart of these processes is the ability of microtubules to do work, a property that derives from their intrinsic dynamic behavior. However, if microtubule dynamics were not properly regulated, it is certain that microtubules alone could not accomplish any of these tasks. In vivo, the regulation of microtubule dynamics is the responsibility of microtubule associated proteins. Among these, we can distinguish several classes according to their function: (1) promotion and stabilization of microtubule polymerization, (2) destabilization or severance of microtubules, (3) functioning as linkers between various structures, or (4) motility-related functions. Here we discuss how the various properties of microtubule-associated proteins can be used to assemble an efficient mitotic apparatus capable of ensuring the bona fide transmission of the genetic information in animal cells. PMID- 15548420 TI - Molecular and functional analysis of the dictyostelium centrosome. AB - The centrosome is a nonmembranous, nucleus-associated organelle that functions not only as the main microtubule-organizing center but also as a cell cycle control unit. How the approximately 100 different proteins that make up a centrosome contribute to centrosome function is still largely unknown. Considerable progress in the understanding of centrosomal functions can be expected from comparative cell biology of morphologically different centrosomal structures fulfilling conserved functions. Dictyostelium is an alternative model organism for centrosome research in addition to yeast and animal cells. With the elucidation of morphological changes and dynamics of centrosome duplication, the establishment of a centrosome isolation protocol, and the identification of many centrosomal components, there is a solid basis for understanding the biogenesis and function of this fascinating organelle. Here we give an overview of the prospective protein inventory of the Dictyostelium centrosome based on database searches. Moreover, we focus on the comparative cell biology of known components of the Dictyostelium centrosome including the gamma-tubulin complex and the homologues of centrin, Nek2, XMAP215, and EB1. PMID- 15548421 TI - Polytene chromosomes: 70 years of genetic research. AB - Polytene chromosomes were described in 1881 and since 1934 they have served as an outstanding model for a variety of genetic experiments. Using the polytene chromosomes, numerous biological phenomena were discovered. First the polytene chromosomes served as a model of the interphase chromosomes in general. In polytene chromosomes, condensed (bands), decondensed (interbands), genetically active (puffs), and silent (pericentric and intercalary heterochromatin as well as regions subject to position effect variegation) regions were found and their features were described in detail. Analysis of the general organization of replication and transcription at the cytological level has become possible using polytene chromosomes. In studies of sequential puff formation it was found for the first time that the steroid hormone (ecdysone) exerts its action through gene activation, and that the process of gene activation upon ecdysone proceeds as a cascade. Namely on the polytene chromosomes a new phenomenon of cellular stress response (heat shock) was discovered. Subsequently chromatin boundaries (insulators) were discovered to flank the heat shock puffs. Major progress in solving the problems of dosage compensation and position effect variegation phenomena was mainly related to studies on polytene chromosomes. This review summarizes the current status of studies of polytene chromosomes and of various phenomena described using this successful model. PMID- 15548422 TI - Insect basic leucine zipper proteins and their role in cyclic AMP-dependent regulation of gene expression. AB - The cAMP-protein kinase A (PKA) pathway is an important intracellular signal transduction cascade that can be activated by a large variety of stimuli. Activation or inhibition of this pathway will ultimately affect the transcriptional regulation of various genes through distinct responsive sites. In vertebrates, the best- characterized nuclear targets of PKA are the cyclic AMP response element-binding (CREB) proteins. It is now well established that CREB is not only regulated by PKA, but many other kinases can exert an effect as well. Since CREB-like proteins were also discovered in invertebrates, several studies unraveling their physiological functions in this category of metazoans have been performed. This review will mainly focus on the presence and regulation of CREB proteins in insects. Differences in transcriptional responses to the PKA pathway and other CREB-regulating stimuli between cells, tissues, and even organisms can be partially attributed to the presence of different CREB isoforms. In addition, the regulation of CREB appears to show some important differences between insects and vertebrates. Since CREB is a basic leucine zipper (bZip) protein, other insect members of this important family of transcriptional regulators will be briefly discussed as well. PMID- 15548423 TI - One-trial aversive learning induces late changes in hippocampal CaMKIIalpha, Homer 1a, Syntaxin 1a and ERK2 protein levels. AB - Most studies regarding altered gene expression after learning are performed using multi-trial tasks, which do not allow a clear discrimination of memory acquisition, consolidation and retrieval. We screened for candidate memory modulated genes in the hippocampus at 3 and 24 h after one-trial inhibitory avoidance (IA) training, using a cDNA array containing 1176 genes. While 33 genes were modulated by training (respect to shocked-only animals), most of them were upregulated (27 genes) and only 6 were downregulated. To confirm and extend these findings, we performed RT-PCRs and analyzed differences in protein levels in rat hippocampus using immunoblot assays. We found several proteins upregulated 24 h after training: extracellular signal-regulated kinase ERK2, Ca2+/calmodulin dependent protein kinase II alpha (CaMKIIalpha), Syntaxin 1a, c-fos and Homer 1a. The total level of none of these proteins were found to be altered when measured 3-h post-training. Several of the mRNAs corresponding to the upregulated proteins were changed at 3 h but not 24 h. Additionally, a number of other candidates were identified for the first time as modulated by learning. The results presented here suggest that single-trial tasks can expose previously unseen differences in dynamic regulation of gene expression after behavioral manipulations, both at the transcriptional and translational levels, and reveal a diversity of gene products modulated by this task, allowing deeper understanding of the molecular basis of memory formation. PMID- 15548424 TI - Role of histaminergic neurons in development of epileptic seizures in EL mice. AB - The EL mouse is an animal model for hereditary temporal lobe epilepsy. When the mice receive weekly vestibular stimulation, e.g., 30 "tosses", 10-15 cm vertically, they start to convulse after 1-2 weeks. The aim of this study was to evaluate the role of the histaminergic neurons in the regulation of seizure development in the EL mice. The obtained results indicated that administration of either histidine, a substrate for histamine synthesis, or metoprine (2,4-diamino 5-(3,4-dichlorophnyl)-6-methyl-pyrimidine), an inhibitor of histamine N methyltransferase (HNMT), retarded the onset of seizure episodes in the mice. The co-administration of histidine and metoprine caused a more marked delay in it. The histamine levels in the brain significantly increased in response to any of these treatments. The intraperitoneal injection of diphenhydramine, a H1 antagonist accelerated the initiation of seizure episodes in the mice, whereas thioperamide, a H3-antagonist caused a delay in the response. There were significant increases in the brain histamine levels upon injection of any of these drugs with concomitant rises in the activity of the histidine decarboxylase (HDC). These results, taken together, suggest that the histaminergic neurons play crucial roles in the development of seizures in the EL mice. They inhibit convulsion in a H1-dependent fashion, while the neurons enhance it in a H3 receptor-mediated way. PMID- 15548425 TI - Oct-3/4 repression accelerates differentiation of neural progenitor cells in vitro and in vivo. AB - Oct-3/4 (Oct-3/Oct-4/POU5F1) is a critical regulator of embryonic stem (ES) cell differentiation, though its role in tissue stem cells that persist in differentiated tissues has not been shown. Here, we show that Oct-3/4 is expressed in neurospheres (NS) composed of neural stem cells and neural progenitor cells and that up- or down-regulation of Oct-3/4 by using adenovirus vectors influences cell fate. Oct-3/4 down-regulation accelerates neuronal differentiation of progenitor cells while its sustained expression prevents neuronal differentiation. Transplantation of neurospheres into the adult rat brain shows that down-regulation of Oct-3/4 promotes differentiation of NS cells in vivo. Our findings indicate that Oct-3/4 is an essential regulator of NS cell differentiation and suggest that the modulation of Oct-3/4 could be a useful tool in clinical application of NS cells. PMID- 15548426 TI - Cyclooxygenase-2 activity contributes to neuronal expression of cyclin D1 after anoxia/ischemia in vitro and in vivo. AB - Cyclooxygenase-2 (COX-2) activity has been implicated in the pathogenesis of neuronal cell death in ischemia and other diseases, but the mechanism by which COX-2 exacerbates cell death is unknown. COX-2 activity is known to induce expression of cyclin D1 in neoplastic cells, and cyclin D1 expression can induce cell death in postmitotic neurons. In the present study, the role of COX-2 and cyclin D1 in neuronal cell death induced by anoxia and ischemia was examined. Treatment with the COX-2 specific inhibitor (NS 398 25 microM) and cyclin D1 inhibitor (flavopiridol 1 microM) increased neuronal survival and inhibited DNA fragmentation after anoxia. NS-398 suppressed anoxia-induced expression of cyclin D1. Flavopiridol inhibited the anoxia-induced increased expression of cyclin D1, but had no effect on COX-2 expression. Treatment with the selective COX-2 inhibitor, SC58125, had no affect on COX-2 expression but partially suppressed cyclin D1 expression in the cortex following middle cerebral artery occlusion in vivo. These results show that COX-2 activity is required for cyclin D1 expression after ischemia in vivo and anoxia in vitro. These data provide support for the hypothesis that cyclin D1 expression is an important mechanism by which COX-2 activity exacerbates ischemic neuronal death. PMID- 15548427 TI - Dishevelled promotes neurite outgrowth in neuronal differentiating neuroblastoma 2A cells, via a DIX-domain dependent pathway. AB - Dishevelled (Dvl) is a cytoplasmic protein involved in the Wnt-Frizzled signaling cascade, which has also been shown to interact with the cytoskeleton in part through inhibition of glycogen synthase kinase 3beta (GSK3beta). Using mouse neuroblastoma 2A (N2A) cells as a model system, we have found that overexpression of Dvl promotes the outgrowth of neurite-like processes, and leads to the induction of a striking, bipolar morphologic phenotype during neuronal differentiation. In contrast, suppression of Dvl expression using isoform specific siRNAs led to an inhibition of neurite outgrowth in these cells. In order to further elucidate the mechanism(s) responsible for this effect, we overexpressed several mutant forms of Dvl in the N2A cells, including deletions in each of the three major functional subdomains of the protein (DeltaDIX, DeltaPDZ, DeltaDEP) and point mutations in the two well-defined interaction motifs within the DIX domain (the actin-binding and vesicle-association elements; K58A and K68A/E69A, respectively). These experiments revealed that the DIX domain (and its vesicle-binding subregion) was essential for Dvl's effect on neurite extension and morphogenesis in N2A cells. In contrast, direct overexpression of a degradation-resistant form of beta-catenin (S37A), or a dominant negative GSK3beta mutant (K85R), had no effect on neurite outgrowth or morphology in neuronally differentiating N2A cells; exposure of cells to a pharmacologic inhibitor of GSK3beta (lithium) also had no effect. Taken together, these results suggest that Dvl induces cytoskeletal and morphologic rearrangements in neuronal differentiating N2A cells through a mechanism that cannot be attributed exclusively to modulation of GSK3beta or beta-catenin activity, but which does depend upon a DIX-domain/vesicle-association-dependent signaling pathway. PMID- 15548428 TI - Catechol-O-methyltransferase gene Val108/158Met polymorphism, and susceptibility to schizophrenia: association is more significant in women. AB - Schizophrenia is a complex disorder with a polygenic inheritance. Catechol-O methyltransferase (COMT) plays a significant role in the regulation of dopaminergic systems. A polymorphism at COMT Val108/158Met has been identified in association with schizophrenia. We examined the allele and genotype association of the COMT Val108/158Met polymorphism of 297 unrelated schizophrenic patients who strictly met DSM-IV criteria for schizophrenia, and 341 healthy controls. We found significant difference in allele and genotype frequencies between schizophrenic patients and controls (chi2=13.030; P=0.001). The allele frequency of the COMT-L was 45.79% in the total schizophrenic patients, and 41.50% in controls. The genotype frequency of the COM-LL was 21.2% in the total schizophrenic patients, and 11.4% in controls (OR=2.085; 95% CI=1.350-3.219; chi2=11.293; P=0.001). With a separate sex analysis, the frequency of the COMT-L allele was moderately distributed in male schizophrenia (chi2=6.177; df=2; P=0.046). The COMT-LL genotype had a 1.818-fold increased risk for schizophrenia (OR=1.818; 95% CI=1.010-3.273; chi2=4.048; P=0.044). The frequency of the COMT-L allele was even more significantly distributed in women schizophrenia (chi2=7.797; df=2; P=0.020). The COMT-LL genotype had remarkably more increased risk for schizophrenia (OR=2.456; 95% CI=1.287-4.687; chi2=7.710; P=0.005). In conclusion, our results provide strong evidence for a role of the COMT-L allele and LL genotype in the etiopathophysiology of schizophrenia with a sexual difference. PMID- 15548429 TI - Identification of differentially expressed genes induced in the rat brain by acetyl-L-carnitine as evidenced by suppression subtractive hybridisation. AB - Acetyl-L-carnitine (ALC) is a molecule widely present in the central nervous system (CNS) formed by the reversible acetylation of carnitine. It acts by stimulating energy metabolism. Reported neurobiological effects of this substance include modulation of brain energy and phospholipid metabolism; cellular macromolecules (including neurotrophic factors and neurohormones); synaptic transmission of multiple neurotransmitters. ALC is of considerable interest for its clinical application in Alzheimer's disease and in the treatment of painful neuropathies. There are experimental data that it affects attention and antagonizes deterioration of ability to learn, improving long-term memory. Moreover, ALC influences nonassociative learning of sensitization type in Hirudo medicinalis. These findings are suggesting that ALC might exert its effects by means of new protein synthesis. ALC or saline solution was injected intraperitoneally each day for 21 days in rats. Poly(A)+ RNAs were isolated from control and treated rat brain. Suppression subtractive hybridisation (SSH) method was applied for the generation of subtracted cDNA libraries and the subsequent identification of differentially expressed transcripts after treatments. The technique generates an equalized representation of differentially expressed genes irrespective of their relative abundance, and it is based on the construction of forward and reverse cDNA libraries that allow the identification of the genes that are regulated or switched off/on after ALC treatment. We identified two modulated genes, the isoform gamma of 14-3-3 protein and a precursor of ATP synthase lipid-binding protein, and one gene switched on by the treatment, the heat shock protein hsp72. PMID- 15548430 TI - Osteopontin (Eta-1) is present in the rat basal ganglia. AB - Osteopontin (OPN) is a secreted glycosylated phosphoprotein that is responsive to oxidative stress and inflammation and controls cytokine production, inducible nitric oxide synthase (iNOS) expression and apoptotic cell death. In this study, we demonstrate the presence of OPN in the rat basal ganglia. Using reverse transcriptase polymerase chain reaction (RT-PCR), OPN cDNA was found in the substantia nigra, and striatum. The presence of OPN mRNA was demonstrated in the same areas of the basal ganglia, using in situ hybridisation. OPN protein was found in the SN, using Western blotting and confirmed by immunohistochemistry. The protein was localised to neurones but not to microglia or astroglia. This is the first report of the presence of OPN in the basal ganglia where it may be involved in the maintenance of neuronal viability. PMID- 15548431 TI - A Purkinje cell specific GoLoco domain protein, L7/Pcp-2, modulates receptor mediated inhibition of Cav2.1 Ca2+ channels in a dose-dependent manner. AB - L7/Pcp-2 is a GoLoco domain protein encoded by a Purkinje cell dendritic mRNA. Although biochemical interactions of GoLoco proteins with Galpha(o) and Galpha(i) are well documented, little is known about effector function modulation resulting from these interactions. The P-type Ca2+ channels might be physiological effectors of L7 because (1) they are the major voltage-dependent Ca2+ channels (VDCC) that modulate Purkinje cell output and (2) they are regulated by G(i/o) proteins. As a first step towards validating this hypothesis and to further understand the possible physiological effect of L7 protein and its two isoforms, we have coexpressed Ca(v)2.1 channels and kappa-opioid receptors (KORs) with varying amounts of L7A or L7B in Xenopus oocytes and measured ionic currents by two-electrode voltage clamping. Without receptor activation L7 did not alter the Ca2+ channel activity. With tonic and weak activation of the receptors, however, the Ca2+ channels were inhibited by 40-50%. This inhibition was enhanced by low, but dampened by high, expression levels of L7A and L7B and differences were observed between the two isoforms. The enhancing effect of L7 was occluded by overexpression of Gbetagamma, whereas the disinhibition was antagonized by overexpression of Galpha(o). We propose that L7 differentially affects the Galpha and Gbetagamma arms of receptor-induced G(i/o) signaling in a concentration dependent manner, through which it increases the dynamic range of regulation of P/Q-type Ca2+ channels by G(i/o) protein-coupled receptors. This provides a framework for designing further experiments to determine how dendritic local fluctuations in L7 protein levels might influence signal processing in Purkinje cells. PMID- 15548432 TI - Human brain endothelium: coexpression and function of vanilloid and endocannabinoid receptors. AB - The arachidonic acid derivative, 2-arachidonoyl-glycerol (2-AG), was initially isolated from gut and brain; it is also produced and released from blood and vascular cells. Many of the 2-AG-induced cellular responses (i.e., neuromodulation, cytoprotection and vasodilation) are mediated by cannabinoid receptors CB1 and CB2. The findings presented here demonstrate the expression of CB1, CB2 and TRPV1 receptors on cerebromicrovascular endothelial cells (HBEC). The expression of TRPV1, CB1 and CB2 receptor mRNA and proteins were demonstrated by RT-PCR and polyclonal antibodies, respectively. The endocannabinoid 2-AG, and other related compounds [anandamide (ANA), methanandamide (m-ANA), N-(4 hydroxyphenyl-arachidonyl-ethanolamide) (AM404) and capsaicin] dose-dependently stimulated Ca2+ influx in HBEC. The selective TRPV1 receptor antagonist (capsazepine), CB1 receptor antagonist (SR141716A) and CB2 receptor antagonist (SR144528) inhibited these responses. The effects of capsaicin, a specific agonist for TRPV1 receptors, were inhibited by capsazepine, but only weakly by CB1 or CB2 receptor antagonists. 2-AG also induced phosphorylation of vasodilator stimulated phosphoprotein (VASP); this response was mediated by VR1 receptors. These studies clearly indicate that 2-AG and other related compounds may function as agonists on VR1 receptors, as well as CB1 and CB2 receptors, and implicated these factors in various HBEC functions. PMID- 15548433 TI - EAU guidelines on chronic pelvic pain. AB - OBJECTIVES: On behalf of the European Association of Urology (EAU) guidelines for diagnosis, therapy and follow-up of chronic pelvic pain patients were established. METHOD: Guidelines were compiled by a working group and based on current literature following a systematic review using MEDLINE. References were weighted by the panel of experts. RESULTS: The full text of the guidelines is available through the EAU Central Office and the EAU website (www.uroweb.org). This article is a short version of this text and summarises the main conclusions from the guidelines on management of chronic pelvic pain. CONCLUSION: A guidelines text is presented including chapters on prostate pain and bladder pain syndromes, urethral pain, scrotal pain, pelvic pain in gynaecological practice, role of the pelvic floor and pudendal nerve, general treatment of chronic pelvic pain and neuromodulation. These guidelines have been drawn up to provide support in the management of the large and difficult group of patients suffering from chronic pelvic pain. PMID- 15548434 TI - Laparoscopic nephroureterectomy for upper urinary tract transitional cell carcinoma: is it better than open surgery? AB - OBJECTIVE: In 1991, laparoscopic nephroureterectomy has been introduced as a treatment option for upper tract transitional cell carcinoma. Based on the review of the current literature and personal experience we want to analyze the actual results of this technique in comparison to open surgery. MATERIALS AND METHODS: We performed a MEDLINE/PubMed search and reviewed the literature on laparoscopic and open nephroureterectomy between 1991 and 2004 (n = 1365 patients) including the results of 45 patients who underwent either laparoscopic (n = 23) or open nephroureterectomy (n = 21) during the same period of time at the Klinikum Heilbronn. Demographic, perioperative and follow-up data were compared. RESULTS: The analysis revealed a slightly longer OR-time (276.6 vs. 220.1 min), and significantly lower blood loss (240.9 vs. 462.9 ml) in the laparoscopic series. No differences of minor (12.9 vs. 14.1%) or major complication rate (5.6 vs. 8.3%) were observed. All nine comparative studies revealed a significant dose reduction of the morphine-equivalents after laparoscopy. In all ten comparative series the hospital stay was shorter after laparoscopy, but only in 6 series the difference was statistically significant. The frequency of bladder recurrence (24.0 vs. 24.7%), local recurrence (4.4 vs. 6.3%), and distant metastases (15.5% vs. 15.2) did not differ significantly in both groups. The actual disease-free two-year survival rates (75.2 vs. 76.2%) were similar. The five-year survival rates averaged 81.2% in the three laparoscopic (n = 113 pat.) and 61% in the ten open series (n = 681 pat.) Six port site metastases were reported in 377 (1.6%) analyzed patients occurring 3 to 12 months following laparoscopy. CONCLUSION: Open radical nephroureterectomy still represents the golden standard for the management of upper tract transitional cell carcinoma, however, laparoscopic radical nephroureterectomy offers the advantages of minimally invasive surgery without deteriorating the oncological outcome. In case of advanced tumors (pT3,N+) open surgery is still recommended. PMID- 15548435 TI - Molecular biomarker in prostate cancer: the role of CpG island hypermethylation. AB - CpG island hypermethylation may be one of the earliest somatic genome alterations to occur during the development of multiple cancers. Recently, aberrant methylation patterns for different tumors have been reported. We present a comprehensive review of the literature describing the role of CpG island hypermethylation of DNA from prostatic tissue and bodily fluids from men with prostate cancer. We reviewed the literature to evaluate CpG island hypermethylation in tissue and bodily fluids of men with primary and metastatic prostate cancer. Additionally, we reviewed the literature with respect to CpG island hypermethylation patterns in other urological malignancies. Using modern analytic methods, CpG island hypermethylation detection can be achieved. In men with prostate cancer, correlations between specific gene regulatory region hypermethylation analyses and Gleason score, pathologic stage and tumor recurrence have been demonstrated. CpG island hypermethylation may serve as a useful molecular biomarker for the detection and diagnosis of patients with prostate cancer. PMID- 15548436 TI - Chemotherapy in hormone refractory prostate cancer: where do we stand? PMID- 15548438 TI - Clinical consequences of screening for prostate cancer: 15 years follow-up of a randomised controlled trial in Sweden. AB - OBJECTIVE: To test the feasibility of a population-based prostate cancer screening programme in general practice and explore the outcome after a 15-year follow-up period. METHODS: From the total population of men aged 50-69 years in Norrkoping (n = 9026) every sixth man (n = 1494) was randomly selected to be screened for prostate cancer every third year over a 12-year period. The remaining 7532 men were treated as controls. In 1987 and 1990 only digital rectal examination (DRE) was performed, in 1993 and 1996 DRE was combined with a test for Prostate-Specific Antigen (PSA). TNM categories, grade of malignancy, management and cause of death were recorded in the South-East Region Prostate Cancer Register. RESULTS: There were 85 (5.7%) cancers detected in the screened group (SG), 42 of these in the interval between screenings, and 292 (3.8%) in the unscreened group (UG). In the SG 48 (56.5%) of the tumours and in the UG 78 (26.7%) were localised at diagnosis (p < 0.001). In the SG 21 (25%) and in the UG 41 (14%) received curative treatment. There was no significant difference in total or prostate cancer-specific survival between the groups. CONCLUSIONS: Although PSA had not been introduced in the clinical practice at the start of the study, we were still able to show that it is possible to perform a long-term population-based randomised controlled study with standardised management and that screening in general practice is an efficient way of detecting prostate cancer whilst it is localised. Complete data on stage, treatment and mortality for both groups was obtained from a validated cancer register, which is a fundamental prerequisite when assessing screening programmes. PMID- 15548437 TI - Weekly docetaxel and vinorelbine (VIN-DOX) as first line treatment in patients with hormone refractory prostate cancer. AB - BACKGROUND: The current study investigated the clinical benefit, the impact on biochemical and objective response and tolerability of weekly docetaxel with vinorelbine (VIN-DOX) in symptomatic patients with hormone refractory prostate cancer (HRPC). METHODS: Patients were treated with docetaxel 25 mg/m2 and vinorelbine 20 mg/m2, intravenously for 6 consecutive weeks followed by a 2 week rest repeatedly until disease progression. Clinical benefit evaluations, based on Karnofsky performance status (KPS) and pain measure, were assessed weekly during therapy. A clinical benefit response was defined as a sustained (> or =4 weeks) improvements in one of these parameters. Changes in prostate-specific antigen (PSA) levels, tumoral response and toxicity also were evaluated. RESULTS: 19 men (median age 68 years), were treated. Overall, 42% of patients achieved a KPS significant change and positive pain response; 47% achieved a 50% or greater reduction in PSA. The objective response rate was observed in 2 of 9 patients with measurable disease (22%). The most important toxicity was neutropenia (Grade 3 = 32%). CONCLUSIONS: The combination of weekly VIN-DOX appears to be feasible. VIN-DOX was found to be associated with improvement in clinical benefit response and biochemical response and well tolerated as first line treatment in HRPC. PMID- 15548439 TI - Systematic development of a guideline for early detection of prostate cancer: the German way in the evidence gap. AB - OBJECTIVE: To develop an evidence-based guideline, helping physcians make prudent decisions about diagnostic care for men wishing to undergo examination for early detection of prostate cancer. METHODS: A guideline development group, comprised of twenty healthcare professionals, including urologists, clinical chemists, pathologists, geriatricians, epidemiologists, technicians and a member of a patient self-help group, systematically gathered, evaluated, and discussed the most recent research available on early detection of prostate cancer. Nominal group technique (NGT) was employed to facilitate the decision-making process. RESULTS: The NGT was sufficient to find a consensus among different medical disciplines in a timely fashion. A standardized guideline, containing a short version for physicians and a standardized patient information booklet, for nation wide use was developed. CONCLUSIONS: Population-based screening is not favoured, instead information should be given to men in their 50-70s, to enable shared decision-making between physician and patient for or against PSA-based early detection of prostate cancer. PMID- 15548440 TI - Skeletal morbidity in men with prostate cancer: quality-of-life considerations throughout the continuum of care. AB - OBJECTIVE: With current treatments, men usually survive many years after being diagnosed with prostate cancer. However, without supportive care, the systemic effects of prostate cancer and therapies such as androgen deprivation therapy (ADT) can undermine skeletal integrity, resulting in skeletal complications that may erode quality of life (QOL). Prostate cancer patients are at risk for fractures from cancer treatment-induced bone loss. In addition, they are also at risk for pathologic fractures, severe bone pain, and other sequelae from bone metastases, which almost invariably occur during the progression of prostate cancer. This review investigates the incidence and pathophysiology of bone loss and skeletal morbidity in prostate cancer patients and reviews available treatment options for maintaining skeletal health throughout the continuum of care for these patients. METHODS: Studies were identified through MEDLINE searches, review of bibliographies of relevant articles, and review of abstracts from national meetings. RESULTS: Several supportive care options are available to prevent generalized and localized bone loss, including calcium and vitamin D supplements and bisphosphonates. Oral calcium and vitamin D supplementation alone, however, appears to be insufficient to prevent bone loss during ADT. Zoledronic acid administered every 3 months during ADT or every 3 to 4 weeks for patients with bone metastases can reverse bone loss and reduce skeletal morbidity, respectively, in patients with prostate cancer. CONCLUSIONS: Skeletal complications contribute to the erosion of QOL in prostate cancer patients. Palliative care can provide important benefits to these patients. Some agents, such as zoledronic acid, may provide skeletal health benefits throughout the course of prostate cancer progression. Further investigations of the QOL impact of these benefits are warranted. PMID- 15548441 TI - Transurethral approach to the distal ureter in nephroureterectomy: transurethral extraction vs. "pluck" technique with long-term follow-up. AB - OBJECTIVES: We retrospectively compared two techniques of transurethral management of the lower ureter in nephroureterectomy. PATIENTS AND METHODS: From August 1992 to December 2003, 34 patients underwent either transurethral detachment of the intramural ureter and cephalad extraction ("pluck"; Group 1, N = 18) or transection of the ureter with subsequent transurethral extraction (Group 2, N = 16). Choice of technique was left to the operating surgeon. All patients with upper tract urothelial carcinoma (TCC) were regularly followed by cystoscopy and abdominal ultrasound. RESULTS: Of the 34 patients, 29 had upper tract TCC. Mean follow-up in these was 44 months (range: 1-129), with 24 (83.8%) over 24 months. On follow-up, 14 bladder tumors (all superficial) occurred in 7 patients (24.1%), but in no case on the scar of the excised ureteral orifice. No extravesical recurrences in the former ureteral bed were found. Of the 29 with upper tract TCC, 19 (65.5%) are alive without disease (median 45 months, range: 6 129), 5 (17.2%) have died with no evidence of disease (median 34 months, range: 20-58), and 4 (13.8%) have died from progressive disease (median 18 months, range: 1-33); 1 patient was lost to follow-up at 34 months with no evidence of disease. Differences between techniques with regard to blood loss, operative time, complications, and oncologic outcome were not significant. CONCLUSION: Both techniques proved technically and oncologically safe. Bladder tumor recurrence rate was in the range reported for classic nephroureterectomy. No extravesical tumor recurrence in the former ureteral bed or on the scar of the resected ureteral orifice occurred. PMID- 15548442 TI - Long-term follow-up of ureteral stump tumors after nephrectomy for benign renal disease. AB - OBJECTIVE: The occurrence of primary carcinoma of the ureteral stump after nephrectomy is rare. In this study, we evaluated the clinical characteristics of ureteral stump tumors after nephrectomy for benign renal disease. METHODS: During a 16-year period, 318 consecutive patients underwent simple nephrectomy for benign renal disease (216 cases) or for donation (102 cases). Eight of these 318 patients diagnosed as having an ureteral stump tumor were treated by ipsilateral ureterectomy with cuff excision of the bladder. Pathologic findings, tumor stages, and clinical characteristics were analyzed. RESULTS: The eight ureteral stump tumors comprised; 6 transitional cell carcinomas (TCCs) and 2 squamous cell carcinomas (SCCs). The mean interval between nephrectomy and ureteral stump tumor diagnosis was 76.5 months. Six of the 8 patients had pyonephrosis and two renal tuberculosis as original renal diseases. Four of the 6 TCCs were stage T1 and 2 stage T2. There was no concomitant bladder tumor at stump tumor diagnosis. Hematuria was the major presenting symptom in 3 of the 8 patients and 4 patients were diagnosed by follow-up imaging study. Two of the 6 ureteral stump TCC patients developed bladder TCC during follow-up. The 5-year survival rate of patients with ureteral stump tumor was 37.5%. T1G1 TCC was associated with a better survival than T2 or G2 TCC. No ureteral stump tumor occurred in cases of donor nephrectomy. CONCLUSION: This study demonstrate, that long-term closed observation is needed to detect ureteral stump tumor, particularly in patients that have undergo nephrectomy for a long-standing inflammatory renal disease such as pyonephrosis or tuberculosis. Hematuria is a major presenting symptom of ureteral stump tumor. However, a follow-up imaging study is also important for ureteral stump tumor detection. The prognosis is poor in cases developing ureteral stump SCC, bladder tumor recurrence, or a high-grade ureteral tumor. PMID- 15548443 TI - Validity of digital rectal examination and serum prostate specific antigen in the estimation of prostate volume in community-based men aged 50 to 78 years: the Krimpen Study. AB - BACKGROUND: In men with symptoms suggestive of BPH, an accurate estimation of the degree of prostate volume enlargement is important for the choice of treatment, and for prediction of treatment effect, the risk of acute urinary retention and the need for surgery. In a community-based population of men, the performance of digital rectal examination (DRE) and serum prostate specific antigen (PSA) is compared to planimetric transrectal ultrasonometry (planimetric TRUS) of the prostate. In this way we search for a practical, reliable, and reproducible alternative to TRUS that can be applied in a primary care setting and in the initial evaluation of men with lower urinary tract symptoms. METHODS: Data were collected from 1688 men aged 50 to 78 years recruited in a population-based study. Measurements included serum PSA, DRE, and planimetric TRUS for the estimation of prostate volume. RESULTS: The AUC values of the receiver-operating curves (ROC) curves for serum PSA as a method for the discrimination of prostate volumes above or below 30, 40 and 50 cc are 0.79, 0.86 and 0.92, respectively. DRE has limited value in the estimation of prostate volume and is only good in identifying very large prostates (>50 cc). CONCLUSIONS: In the general male population serum PSA performs reasonably well compared with planimetric TRUS, and better than DRE, in estimating whether prostate volume is greater or smaller than 30, 40 or 50 cc. Serum PSA is an acceptable alternative method to estimate the degree of prostatic enlargement in clinical settings where TRUS is not available and when prostate cancer has been excluded. PMID- 15548444 TI - Effect of NIH-IV prostatitis on free and free-to-total PSA. AB - OBJECTIVE: To examine the effect of asymptomatic prostatic inflammation (NIH category IV prostatitis) on total PSA (tPSA), free serum PSA (fPSA) and the ratio of free-to-total prostate specific antigen (%fPSA). The role of free and %fPSA as a diagnostic tool for distinguishing between cancer and non-malignant diseases of the prostate was also investigated. MATERIAL AND METHODS: In a retrospective study 1090 prostate biopsies performed between January 2000 and September 2003 were evaluated and the levels of serum total and free PSA as well as the f/tPSA ratio were determined in samples obtained immediately before biopsy. 404 patients with full clinical and histological records were included in the study. All patients underwent 6 or 8 core primary prostate needle biopsies. RESULTS: A total of 404 patients were included in the analysis. 100 prostate cancer (PCa) (24.8%), 137 NIH-IV prostatitis (33.9%) and 143 patients with benign prostatic hyperplasias (BPH) (35.4%) were identified. 24 (5.9%) patients presented with both PCa and prostatitis on histology and were excluded from further analysis. The mean (median) levels of tPSA, fPSA and %fPSA were 11.94 ng/ml (8.0), 1.31 ng/ml (1.07) and 0.15 (0.14) for NIH-IV prostatitis; 11.94 ng/ml (8.35), 1.54 ng/ml and 0.13 (0.11) for prostate cancer; and 8.19 ng/ml (7.0), 1.48 ng/ml (1.03) and 0.18 (0.15) for BPH. No significant difference was found in tPSA levels between PCa and prostatitis (p = 0.32), while the difference in tPSA levels between PCa and BPH was significant (p = 0.007). Free PSA alone had no diagnostic power in distinguishing PCa from prostatitis (p = 0. 37) and BPH (p = 0. 61). By contrast, the f/tPSA ratio showed significant between-group differences (PCa versus prostatitis (p = 0. 011), PCa versus BPH (p = 0.0001). CONCLUSIONS: Chronic asymptomatic prostatitis NIH category IV has similar effects on total PSA and free PSA levels in serum as PCa. fPSA alone cannot distinguish prostate cancer from non-malignant inflammatory disease of the prostate. The ratio of free-to-total PSA is significantly different in PCa and NIH category IV prostatitis. PMID- 15548445 TI - Vessels in benign prostatic hyperplasia contain more binding sites for endostatin than vessels in normal prostate tissue. AB - OBJECTIVE: The angiogenic phenotype is an effect of a net balance of angiogenic and anti-angiogenic factors. Endostatin is one of a group of recently described matrix-derived inhibitors of tumour angiogenesis that have acquired increasing significance for tumour treatment. Endostatin's anti-angiogenic mechanism and the causes of its heterogenic influence on various tissue types have not yet been defined. METHODS: We investigated the variations in endostatin's binding behaviour to vessels in benign prostatic hyperplasia (BPH) compared to endostatin binding to vessels in normal prostate tissue. Biotinylated endostatin was used and was detected using Extravidin CY3. RESULTS: In BPH 89.12% +/- 10.72% of vessels showed endostatin binding. This was significantly more than observed for vessels in normal prostate tissue (1.66% +/- 1.66%). CONCLUSION: The strongly proliferative tissue of BPH may be a growth-limited cause of significantly more endostatin binding sites. The investigation indicates endostatin as a possible new target for BPH treatment. PMID- 15548446 TI - Early conceptual and linguistic development of a patient and partner treatment satisfaction scale (TSS) for erectile dysfunction. AB - OBJECTIVE: To describe the early development of a pluri-language self-report questionnaire to assess male patients and their female partners' satisfaction with drug treatment for erectile dysfunction (ED). METHODS: This first development phase proceeded in several parts. Item generation followed literature review, hypothesized characteristics of the drug and in-depth interviews with patients and their partners. Perceptions and feelings related to ED and patients' expectations of treatment were explored. Items were generated simultaneously in 5 languages (American English, Canadian French, English, French and German). Content and face validity were empirically assessed by interviews with a few patients and partners in each country. Conceptual equivalence between languages was ascertained. RESULTS: The final content domains included satisfaction with: sexual spontaneity, quality of erection, quality of ejaculation, sexual pleasure, orgasm, confidence, reliability of treatment, side effects, convenience, overall satisfaction, conformity to treatment expectations and intent to continue use of drug. Cognitive debriefing with patients and partners found few issues with comprehension, however some words were considered problematic. The simultaneous development for the different languages allowed adaptation of the content at this stage and ensured consistency of all language versions. The final questionnaire consisted of 4 modules: unmedicated patient, medicated patient, unmedicated partner, and medicated partner modules. The questionnaire was then linguistically validated into 15 additional languages for further psychometric validation. CONCLUSIONS: The Treatment Satisfaction Scale (TSS) is a multi-facetted measure of patients' and partners' satisfaction with their sexual life relating to erectile dysfunction and intended for prospective use. Its simultaneous development for a variety of countries and languages has fostered true item equivalence across language versions. However, further work is needed to validate the TSS psychometrically, including identification of domains, test responsiveness and determination of appropriate scoring prior to its clinical use. PMID- 15548447 TI - Comparison of the efficacy of tolterodine and oxybutynin in different urodynamic severity grades of idiopathic detrusor overactivity. AB - OBJECTIVE: To compare the efficacy of tolterodine and oxybutynin in the treatment of specific, according to their urodynamic grade of severity, populations with overactive detrusor. METHODS: In this open, randomized, two-way crossover study 128 women with urodynamically confirmed, idiopathic detrusor overactivity were recruited. Patients were categorized in 4 grades of severity groups, according to the characteristics of the first overactive detrusor contraction during filling cystometrogram: high volume-low pressure (grade-group I), high volume-high pressure (grade-group II), low volume-low pressure (grade-group III) and low volume-high pressure (grade-group IV). The primary outcome measure was average volume of voided urine per micturition. RESULTS: 107 patients successfully completed the study protocol and were included in the analyses: 40 in group IV, 36 in III, 25 in II and 6 in group I. In groups IV and III both oxybutynin and tolterodine significantly increased the average volume of voided urine per micturition but the differences between the drugs were not significant (p > 0.05). In group II neither of the drugs achieved significant changes in the outcome measure (p > 0.05). CONCLUSIONS: Tolterodine and oxybutynin are clinically equipotent in treating detrusor overactivity in specific severity groups of patients, although urodynamic effects are somewhat different. PMID- 15548448 TI - Lack of ultrastructural detrusor changes following endoscopic injection of botulinum toxin type a in overactive neurogenic bladder. AB - INTRODUCTION: Endoscopical injections of Botulinum toxin type A into the detrusor muscle are gaining clinical acceptance in the treatment of neurogenic detrusor overactivity. Structural effects of Botulinum toxin type A are only known from studies on striated muscles, where a widespread nerve sprouting occurs temporarily. The aim of this study was to evaluate the ultrastructural effects of Botulinum toxin type A injections on the human detrusor. MATERIAL AND METHODS: 30 detrusor biopsies were obtained from 24 patients with neurogenic detrusor overactivity. Patients were divided into two groups: Group I included 13 biopsies from patients before the first Botulinum toxin type A injection. Group II included 6 biopsies from patients within 3 months after the first injection and 11 biopsies at the time of decreasing efficacy of Botulinum toxin type A. The biopsies were processed by standard procedure for detailed electron microscopic study and evaluated by 2 examiners without prior knowledge of clinical/urodynamic data. RESULTS: No statistically significant detrusor changes have been found concerning muscle cell fascicle structure (p = 0.445), width of intercellular space (p = 0.482) and number/kind of muscle cell junctions (p = 0.443). A median of 70% of intrinsic axon terminals presented with signs of degeneration in group I, a median of 66% in group II (p = 0.840). Out of 309 evaluated axon terminals in both groups, 1 sprouting axon was found in group I, 3 sprouting axons in group II (p = 0.864). Specimen from group I and group II showed only limited collagen deposits within the detrusor. No changes in the ultrastructure of the detrusor have been observed in those biopsies obtained before and after the Botulinum toxin type A injection of the same patient. CONCLUSION: This study verifies our earlier report of severe intrinsic axon degeneration in the detrusor of patients with neurogenic detrusor overactivity. It also shows nearly no structural differences of the detrusor before and after Botulinum toxin type A injections. Contrary to reports of striated muscle, axonal sprouting within the detrusor was very limited after Botulinum toxin type A injections indicating pathophysiologically different reactions to the toxin either between striated muscle and smooth muscle or between different treated diseases. PMID- 15548449 TI - Immunohistochemical evidence of vanilloid receptor 1 in normal human urinary bladder. AB - PURPOSE: Experimental and clinical evidences have shown the importance of the vanilloid receptor 1 (TRPV1) in the lower urinary tract. In humans, this receptor has been detected in nerve endings of primary sensory neurons, smooth muscle and connective tissue cells and in the rat also in the urothelium. The aim of this study is to identify, by immunohistochemistry, the cell types expressing TRPV1 in the human urinary bladder. MATERIAL AND METHODS: Specimens, obtained from normal urinary bladder by multiple biopsy and from ureter at the time of radical nefrectomy for renal cell carcinoma, were fixed and frozen. Full-thickness sections were processed for light and fluorescence microscopes. To label the TRPV1, three polyclonal antibodies were used: the anti-capsaicin receptor, the anti-VR1 (N-15) and the anti-VR1 (C-15). RESULTS: Urothelium, smooth muscle cells, mast cells and endothelium were labelled and the labelling was intracytoplasmatic. In the urothelial cells, the labelling was slightly granular. In the bladder urothelium, the superficial cells were more intensely stained than the basal and club-shaped cells. VR1-positive nerve fibers were seen running single and/or in groups in the sub-urothelium and as single varicose fibers in the muscle coat, and VR1-positive nerve endings in the urothelium. CONCLUSION: The present findings provide the evidence of the presence of TRPV1 on normal human urothelium where it could have important implications in the mechanism of action of intravesical vanilloids (capsaicin and resiniferatoxin). PMID- 15548450 TI - Increased expression of connexin 43 in the overactive neurogenic detrusor. AB - PURPOSE: The cellular mechanisms involved in unstable uncontrolled detrusor contractions in the human bladder remain unknown. One hypothesis, based on electron microscopical observations and animal studies, is that gap junctions are present in the human detrusor and are increased in patients with detrusor overactivity. Thus intercellular electrical coupling between adjacent detrusor cells would be increased allowing electrical activity to spead more easily within the detrusor muscle mass and more readily generate significant contractions. The aim of this study was to prove this hypothesis. MATERIAL AND METHODS: Detrusor biopsies have been obtained from videourodynamically evaluated patients with neurogenic detrusor overactivity (NDO) (n = 19) [group I] and from patients with stress urinary incontinence (SUI) with stable, non-obstructed detrusors (n = 5) [group II] serving as controls. Specimens were fixed, paraffin embedded, sectioned, stained with a monoclonal connexin 43 antibody and evaluated by two blinded examiners using a semiquantitative scale. Connexin 43 mRNA levels were evaluated using quantitative RT-PCR with primers for connexin 43 and for 18S rRNA. The results were correlated with the patients' groups. RESULTS: Connexin 43 could be identified in cross sections of every detrusor biopsy. In all biopsies from patients with NDO a widespread presence of connexin 43 staining was observed, whereas only a limited presence of connexin 43 staining was observed in the specimen from patients with SUI. Connexin 43 mRNA levels within the detrusor were 3.7 fold higher in the NDO group than in the SUI group (p = 0.017). CONCLUSIONS: A limited amount of gap junctions seems to be present in every detrusor independently from its urodynamic stability or instability. But there is a significant increase in connexin 43 protein and RNA levels in patients with neurogenic detrusor overactivity underlining their role in intercellular electrical coupling. PMID- 15548451 TI - Predicting the efficiency of UAG translational stop signal through studies of physicochemical properties of its composite mono- and dinucleotides. AB - In this study, we explored the problem of predicting the UAG stop-codon read through efficiency. The reported nucleotide sequences were first converted into physicochemical property vectors before being presented to a machine learning algorithm. Two sets of physicochemical properties were applied: one for mononucleosides (in terms of steric bulk, hydrophobicity and electronics) and another for dinucleotides. To the best of our knowledge, this is the first report of how dinucleotides are converted into principle components derived from NMR chemical shift data. A few efficiency prediction models were then derived and a comparison between mononucleoside and dinucleotide-based models was shown. In the derived models, the coefficients of these property based predictors lend themselves to bio-physical interpretations, an advantage which is demonstrated in this study via a prediction model based on the steric bulk factor. Although it is quite simple, the steric bulk factor model explained well the effect of sequence variations surrounding the amber stop codon and the tRNA bearing UCCU anticodon. We further proposed new alternatives at position -1 and +4 of a UAG stop codon sequence to enhance the readthrough efficiency. This research may contribute to a better understanding of the readthrough mechanisms and may also help to study the normal translation termination process. PMID- 15548452 TI - Scale-free networks versus evolutionary drift. AB - Recent studies of properties of various biological networks revealed that many of them display scale-free characteristics. Since the theory of scale-free networks is applicable to evolving networks, one can hope that it provides not only a model of a biological network in its current state but also sheds some insight into the evolution of the network. In this work, we investigate the probability distributions and scaling properties underlying some models for biological networks and protein domain evolution. The analysis of evolutionary models for domain similarity networks indicates that models which include evolutionary drift are typically not scale free. Instead they adhere quite closely to the Yule distribution. This finding indicates that the direct applicability of scale-free models in understanding the evolution of biological network may not be as wide as it has been hoped for. PMID- 15548453 TI - A hidden Markov model with molecular mechanics energy-scoring function for transmembrane helix prediction. AB - A range of methods has been developed to predict transmembrane helices and their topologies. Although most of these algorithms give good predictions, no single method consistently outperforms the others. However, combining different algorithms is one approach that can potentially improve the accuracy of the prediction. We developed a new method that initially uses a hidden Markov model to predict alternative models for membrane spanning helices in proteins. The algorithm subsequently identifies the best among models by ranking them using a novel scoring function based on the folding energy of transmembrane helical fragments. This folding of helical fragments and the incorporation into membrane is modeled using CHARMm, extended with the Generalized Born surface area solvent model (GBSA/IM) with implicit membrane. The combined method reported here, TMHGB significantly increases the accuracy of the original hidden Markov model-based algorithm. PMID- 15548454 TI - Classifying G-protein coupled receptors with bagging classification tree. AB - G-protein coupled receptors (GPCRs) play a key role in different biological processes, such as regulation of growth, death and metabolism of cells. They are major therapeutic targets of numerous prescribed drugs. However, the ligand specificity of many receptors is unknown and there is little structural information available. Bioinformatics may offer one approach to bridge the gap between sequence data and functional knowledge of a receptor. In this paper, we use a bagging classification tree algorithm to predict the type of the receptor based on its amino acid composition. The prediction is performed for GPCR at the sub-family and sub-sub-family level. In a cross-validation test, we achieved an overall predictive accuracy of 91.1% for GPCR sub-family classification, and 82.4% for sub-sub-family classification. These results demonstrate the applicability of this relative simple method and its potential for improving prediction accuracy. PMID- 15548455 TI - Fast and high precision algorithms for optimization in large-scale genomic problems. AB - There are several very difficult problems related to genetic or genomic analysis that belong to the field of discrete optimization in a set of all possible orders. With n elements (points, markers, clones, sequences, etc.), the number of all possible orders is n!/2 and only one of these is considered to be the true order. A classical formulation of a similar mathematical problem is the well known traveling salesperson problem model (TSP). Genetic analogues of this problem include: ordering in multilocus genetic mapping, evolutionary tree reconstruction, building physical maps (contig assembling for overlapping clones and radiation hybrid mapping), and others. A novel, fast and reliable hybrid algorithm based on evolution strategy and guided local search discrete optimization was developed for TSP formulation of the multilocus mapping problems. High performance and high precision of the employed algorithm named guided evolution strategy (GES) allows verification of the obtained multilocus orders based on different computing-intensive approaches (e.g., bootstrap or jackknife) for detection and removing unreliable marker loci, hence, stabilizing the resulting paths. The efficiency of the proposed algorithm is demonstrated on standard TSP problems and on simulated data of multilocus genetic maps up to 1000 points per linkage group. PMID- 15548458 TI - Assessment of therapists' and patients' personality: relationship to therapeutic technique and outcome in brief dynamic psychotherapy. AB - My first focus of this study was to explore therapists' personal characteristics as predictors of the proportion of interpretation in brief dynamic psychotherapy (N=39; maximum 40 sessions). In this study, I used data from the Norwegian Multicenter Study on Process and Outcome of Psychotherapy (1995). The main finding was that therapists who had experienced good parental care gave less interpretation (28% variance was accounted for). Therapists who had more negative introjects used a higher proportion of interpretation (16% variance was accounted for). Patients' pretreatment characteristics were not predictive of therapists' use of interpretation. The second focus was to investigate the impact of therapists' personality and the proportion of interpretation on the development of patients' maladaptive defensive functioning over the course of therapy. Better parental care and less negative introjects in therapists were associated with a positive influence and accounted for 5% variance in the reduction of patients' maladaptive defense. PMID- 15548459 TI - Attachment predicts treatment completion in an eating disorders partial hospital program among women with anorexia nervosa. AB - The goal of this study was to examine if attachment theory can provide a framework for understanding treatment completion in an eating disorders partial hospital program among women with anorexia nervosa (AN). Attachment was measured using the Attachment Styles Questionnaire (Feeney, Noller, & Hanrahan, 1994). As hypothesized, self-reports of high avoidant attachment predicted noncompletion of treatment for those with AN binge-purge subtype (ANB). However, this relationship did not emerge for those with AN restricting subtype (ANR). Also as hypothesized, self-reports of high anxious attachment predicted completing treatment for those with ANB but not for those with ANR. For completers with ANB and ANR, the program was helpful in increasing body weight and lowering drive for thinness, body dissatisfaction, interpersonal problems, and depression. Attachment avoidance, characterized by devaluing one's need for relationships, may be a contraindication for group-based partial hospital treatment of ANB. Attachment anxiety, characterized by high preoccupation with relationships, may facilitate remaining in treatment for those with ANB. PMID- 15548460 TI - Validity of the inventory of interpersonal problems for predicting treatment outcome: an investigation with the Pennsylvania Practice Research Network. AB - In this study, we examined the relationship between treatment outcome and variables from the Inventory of Interpersonal Problems Circumplex scales (IIP-C; Horowitz, Alden, Wiggins, & Pincus, 2000) in the Pennsylvania Psychological Association's Practice Research Network (PRN; Borkovec, Echemendia, Ragusea, & Ruiz, 2001). The PRN was a naturalistic observation treatment outcome study conducted with clinicians who were providing outpatient therapy. Assessment instruments, including the Compass Assessment System (Howard, Brill, Lueger, O'Mahoney, & Grissom, 1993; Sperry, Brill, Howard, & Grissom, 1996) and the IIP C, were used to assess outcome at the 7th session (N=73) and at termination (N=42). Significant associations were identified between seventh-session outcome and most of the IIP variables. Only IIP elevation and amplitude were related to termination outcome. Elevation, amplitude, and hostile submissive problems were related to treatment length. Ad hoc analyses indicated that the IIP elevation fully mediated the relationships between interpersonal problems and seventh session outcome but not the relationship between amplitude and outcome. We discuss the results in relation to the external validity of the IIP. PMID- 15548461 TI - Interpersonal problems and outcome in outpatient psychotherapy: findings from a long-term longitudinal study in Germany. AB - We used a comprehensive longitudinal data set from Germany to examine trajectories of symptom distress depending on interpersonal problems at study intake measured via the Inventory of Interpersonal Problems-64 (IIP-64; Horowitz, Strau, & Kordy, 1994). Participants (N=622) underwent mid- or long-term outpatient psychotherapy (either psychodynamically oriented psychotherapy, cognitive behavioral therapy, or analytic psychotherapy). Data comprises up to 5 assessments during a 2-year period and was analyzed via hierarchical linear modeling. In the analytic psychotherapy subgroup, initial symptom level was higher in submissive patients. Initial interpersonal problems were not predictive of the rate of symptom change during therapy. Only in psychodynamic treatments, low affiliation positively affected treatment outcome. Interpersonal problems at intake were not related to the number of utilized sessions and utilization rate across treatment subgroups. We discuss the findings and outline future research topics. PMID- 15548462 TI - Self-directed affiliation and autonomy across acute and continuation phase cognitive therapy for recurrent depression. AB - Using Benjamin's (2000) Structural Analysis of Social Behavior, we evaluated change in "self-directed" affiliation and autonomy and prediction of treatment response and relapse/recurrence among adult outpatients with recurrent major depressive disorder consenting to acute phase cognitive therapy (A-CT; Beck, Rush, Shaw, & Emery, 1979; N=156); A-CT responders randomized (N=84) to 8 months of continuation phase cognitive therapy (C-CT; Jarrett, 1989; Jarrett et al., 1998; Jarrett & Kraft, 1997) or assessment-only control; and C-CT and control patients entering a 16-month, assessment-only follow-up (N=74). Self-directed affiliation and autonomy increased after A-CT, and C-CT further increased affiliation and autonomy. Affiliation and autonomy did not predict A-CT response, but lower affiliation and higher autonomy pre-A-CT predicted relapse/recurrence post-A-CT. We discuss potential clinical implications of these results and present case examples to illustrate patterns of change. PMID- 15548463 TI - Relational measures in psychotherapy research on process and outcome: a commentary on the studies. AB - Using Benjamin's (2000) Structural Analysis of Social Behavior, we evaluated change in "self-directed" affiliation and autonomy and prediction of treatment response and relapse/recurrence among adult outpatients with recurrent major depressive disorder consenting to acute phase cognitive therapy (A-CT; Beck, Rush, Shaw, & Emery, 1979; N=156); A-CT responders randomized (N=84) to 8 months of continuation phase cognitive therapy (C-CT; Jarrett, 1989; Jarrett et al., 1998; Jarrett & Kraft, 1997) or assessment-only control; and C-CT and control patients entering a 16-month, assessment-only follow-up (N=74). Self-directed affiliation and autonomy increased after A-CT, and C-CT further increased affiliation and autonomy. Affiliation and autonomy did not predict A-CT response, but lower affiliation and higher autonomy pre-A-CT predicted relapse/recurrence post-A-CT. We discuss potential clinical implications of these results and present case examples to illustrate patterns of change. PMID- 15548464 TI - Rorschach assessment of changes following psychotherapy: a meta-analytic review. AB - I examined Rorschach assessment of personality changes following psychotherapy. I conducted a comprehensive literature search to find all studies using the Rorschach method at least twice for the same participant in connection with psychotherapy. I conducted meta-analyses for 38 samples, and I performed regression analyses to identify moderating factors. Across all Rorschach scores, the total weighted sample effect size was r = .26, and nearly half the variables obtained effect sizes higher than .30. Several moderating factors were found. Most important, effect sizes increased with longer and more intensive therapy. More concern for interscorer reliability was associated with larger effect sizes, whereas a higher degree of scorer blinding was associated with smaller effect size magnitudes. Predicted levels of change based on the regression models indicated substantial increases in effect size with longer therapies. The data indicate that many elements in the Rorschach are valid indicators of change despite the poor reputation the method has acquired within psychotherapy research. PMID- 15548465 TI - Rorschach comprehensive system variables in relation to assessing dynamic capacity and ego strength for psychodynamic psychotherapy. AB - Few studies have concerned the relation between Comprehensive System (CS; Exner, 1991, 1993, 2003) Rorschach variables, and suitability for psychodynamic psychotherapy. In this study, I predicted correlations between 17 rationally selected CS variables and suitability for psychotherapy as represented by ratings of Dynamic Capacity and Ego Strength. The participants were 52 psychotherapy applicants between 20 and 57 years old. The ratings were performed after clinical interviews and decisions concerning selection of the applicants. EA, FC, Blends, Zf, and MQo correlated positively and YFY negatively with Dynamic Capacity ratings, whereas EA, FC, and Blends correlated positively and YFY and F% negatively with Ego Strength ratings. EA, FC, YFY, Zf, MQo, and F% also differed between the applicants selected and those not selected for psychotherapy. Considering earlier research, the results for Blends, Zf, and F% were interpreted as replications and for MQo and EA as showing satisfactory consistency. PMID- 15548466 TI - Rorschach personality predictors of weight loss with behavior modification in obesity treatment. AB - Weight loss outcome for 49 obese patients after 6 months treatment in a behavior modification program was related to Rorschach personality characteristics according to the Comprehensive System (Exner, 2003), also including the Rorschach Oral Dependency Scale (Bornstein, 1996; Masling & Rabie, 1967). Less weight loss was predicted by signs of perceptual and cognitive distortions indicated by the Schizophrenia Index. More weight loss was predicted by Food Contents, suggesting a food preoccupation and possibly a benign dependency orientation. Patients with a focus on food and dependent needs could benefit above all from the treatment program implying learning how to handle food and eating in a supportive setting, as evidenced by more weight loss. Distortions in perception and cognition could constitute more profound difficulties in weight reduction important to recognize in obesity treatment. PMID- 15548467 TI - Personality and symptom change in treatment-refractory inpatients: evaluation of the phase model of change using Rorschach,TAT, and DSM-IV Axis V. AB - In this study, we examined global treatment outcomes during 16 months of intensive, psychodynamic treatment for 77 inpatients suffering from treatment refractory disorders. Hypotheses based on the phase model of treatment change (Howard, Lueger, Maling, & Martinovich, 1993; Howard, Moras, Brill, Martinovich, & Lutz, 1996) were supported in the study results. Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) Axis V scales assessing behavioral functioning demonstrated large and medium effect size change, whereas stable, enduring personality functioning assessed by psychoanalytic Rorschach scales and the Social Cognition and Object Relations Scale (Westen, 1995) for the Thematic Apperception Test (Murray, 1943) demonstrated small and medium effect size change. We also report assessment of reliable change index and clinical significance. The ecological validity of Rorschach measures is supported by significant validity coefficients (in the hypothesized directions) between implicit measures of personality functioning and behavioral ratings. PMID- 15548468 TI - Monitoring psychotherapy with performance-based measures of personality functioning. AB - In this commentary, I review a meta-analysis and three original research reports concerning the Rorschach (Exner, 2003; Rorschach, 1921/1942) and the Thematic Apperception Test (TAT; Murray, 1943) assessment in psychological treatment planning and outcome evaluation. The information in these four articles bears witness to the potential utility of performance-based personality assessment measures for this purpose. The strengths and limitations of the articles suggest several guidelines for future research designed to examine this Rorschach and TAT application including an emphasis on effectiveness studies, longitudinal data, integrated independent variables, observable dependent variables, sophisticated data analysis combining nomothetic and idiographic presentation, and the incremental contribution of performance-based measures to psychotherapy-related personality assessment. PMID- 15548469 TI - The development of therapeutic alliance during psychological assessment: patient and therapist perspectives across treatment. AB - We examined the impact of patient- and therapist-rated alliance developed during psychological assessment on the subsequent alliance measured early and late in formal psychotherapy. We hypothesized that a working alliance developed during psychological assessment conducted from a collaborative therapeutic model of assessment (TMA; Finn & Tonsager, 1992, 1997; Fischer, 1994) between the patient and therapist would carry into formal psychotherapy. We also hypothesized that alliance for those patients receiving a TMA would be significantly greater than patients receiving psychological testing as usual. To test this hypothesis, we administered the Combined Alliance Short Form-Patient Version (Hatcher & Barends, 1996) and the Combined Alliance Short Form-Therapist Version (Hatcher, 1999) to a sample of outpatients and their therapists at the end of the assessment feedback session, early, and late in psychotherapy. The hypotheses were supported as alliance scales rated at the assessment feedback session demonstrated positive and significant relationships with alliance throughout formal psychotherapy and in relation to a control group. The clinical utility and research implications of these findings are discussed. PMID- 15548470 TI - Quality of object relations as a moderator of the relationship between pattern of alliance and outcome in short-term individual psychotherapy. AB - In this study, we investigated the personality variable quality of object relations (QOR) as a moderator of the relationship between the pattern of the therapeutic alliance and treatment outcome in two forms (interpretive, supportive) of short-term individual psychotherapy. In a sample of 72 psychiatric outpatients who completed interpretive therapy, QOR emerged as a moderator for the outcome factor general symptomatology and dysfunction. For high-QOR patients, an increasing level of alliance was directly related to benefit, whereas for low QOR patients, a decreasing level of alliance was directly related to benefit. An explanation for these findings emphasized the importance of patients repeating their typical pattern of maladaptive interpersonal behavior in the therapy sessions in the context of the therapist working with the transference. In a sample of 72 psychiatric outpatients who completed supportive therapy, QOR did not emerge as a moderator. These findings in combination with evidence from previous studies suggest that QOR should be investigated as a moderator variable in future studies of short-term psychotherapy. PMID- 15548472 TI - Thematic review series: the immune system and atherogenesis. Recent insights into the biology of macrophage scavenger receptors. AB - Scavenger receptors were originally defined by their ability to bind and internalize modified lipoproteins. Macrophages express at least six structurally different cell surface receptors for modified forms of LDL that contribute to foam cell formation in atherosclerosis. In addition to their role in the pathology of atherosclerosis, macrophage scavenger receptors, especially SR-A, play critical roles in innate immunity, apoptotic cell clearance, and tissue homeostasis. In this review, we highlight recent advances in understanding the biology of macrophage scavenger receptors as pattern recognition receptors for both infectious nonself (pathogens) and modified self (apoptotic cells and modified LDL). We critically evaluate the potential of scavenger receptors and their ligands as targets for therapeutic intervention in human disease. PMID- 15548473 TI - Antiepileptic drugs in psychiatry. PMID- 15548474 TI - Clear indications of emotion depend on vivid stimuli. PMID- 15548475 TI - Using PET to identify carotid occlusion patients at high risk of subsequent stroke: further insights. PMID- 15548476 TI - Variations in care and outcome in the first year after stroke: a Western and Central European perspective. PMID- 15548477 TI - Does the risk of developing dementia increase with the number of episodes in patients with depressive disorder and in patients with bipolar disorder? AB - OBJECTIVE: Several findings suggest that some patients with depressive or bipolar disorder may be at increased risk of developing dementia. The present study aimed to investigate whether the risk of developing dementia increases with the number of affective episodes in patients with depressive disorder and in patients with bipolar disorder. METHODS: This was a case register study including all hospital admissions with primary affective disorder in Denmark during 1970-99. The effect of the number of prior episodes leading to admission on the rate of readmission with a diagnosis of dementia following the first discharge after 1985 was estimated. A total of 18,726 patients with depressive disorder and 4248 patients with bipolar disorder were included in the study. RESULTS: The rate of a diagnosis of dementia on readmission was significantly related to the number of prior affective episodes leading to admission. On average, the rate of dementia tended to increase 13% with every episode leading to admission for patients with depressive disorder and 6% with every episode leading to admission for patients with bipolar disorder, when adjusted for differences in age and sex. CONCLUSION: On average, the risk of dementia seems to increase with the number of episodes in depressive and bipolar affective disorders. PMID- 15548478 TI - From orthostatic hypotension to Shy-Drager syndrome. PMID- 15548479 TI - Recognition of emotion from moving facial and prosodic stimuli in depressed patients. AB - BACKGROUND: It has been suggested that depressed patients have a "negative bias" in recognising other people's emotions; however, the detailed structure of this negative bias is not fully understood. OBJECTIVES: To examine the ability of depressed patients to recognise emotion, using moving facial and prosodic expressions of emotion. METHODS: 16 depressed patients and 20 matched (non depressed) controls selected one basic emotion (happiness, sadness, anger, fear, surprise, or disgust) that best described the emotional state represented by moving face and prosody. RESULTS: There was no significant difference between depressed patients and controls in their recognition of facial expressions of emotion. However, the depressed patients were impaired relative to controls in their recognition of surprise from prosodic emotions, judging it to be more negative. CONCLUSIONS: We suggest that depressed patients tend to interpret neutral emotions, such as surprise, as negative. Considering that the deficit was seen only for prosodic emotive stimuli, it would appear that stimulus clarity influences the recognition of emotion. These findings provide valuable information on how depressed patients behave in complicated emotional and social situations. PMID- 15548481 TI - Greater syntactic impairments in native language in bilingual Parkinsonian patients. AB - OBJECTIVE: To investigate the presence of syntactic impairments in native language in Parkinson's disease. METHODS: Twelve bilingual patients, with Friulian as their first language (L1) and Italian as their second (L2), with Parkinson's disease and 12 normal controls matched for age, sex, and years of schooling, were studied on three syntactic tasks. RESULTS: Patients with Parkinson's disease showed a greater impairment of L1 than L2. CONCLUSIONS: These findings provide evidence of greater basal ganglia involvement in the acquisition and further processing of grammar in L1 v L2 possibly due to a major involvement of procedural memory in representing L1 grammar. PMID- 15548480 TI - Mucuna pruriens in Parkinson's disease: a double blind clinical and pharmacological study. AB - BACKGROUND: The seed powder of the leguminous plant, Mucuna pruriens has long been used in traditional Ayurvedic Indian medicine for diseases including parkinsonism. We have assessed the clinical effects and levodopa (L-dopa) pharmacokinetics following two different doses of mucuna preparation and compared them with standard L-dopa/carbidopa (LD/CD). METHODS: Eight Parkinson's disease patients with a short duration L-dopa response and on period dyskinesias completed a randomised, controlled, double blind crossover trial. Patients were challenged with single doses of 200/50 mg LD/CD, and 15 and 30 g of mucuna preparation in randomised order at weekly intervals. L-dopa pharmacokinetics were determined, and Unified Parkinson's Disease Rating Scale and tapping speed were obtained at baseline and repeatedly during the 4 h following drug ingestion. Dyskinesias were assessed using modified AIMS and Goetz scales. RESULTS: Compared with standard LD/CD, the 30 g mucuna preparation led to a considerably faster onset of effect (34.6 v 68.5 min; p = 0.021), reflected in shorter latencies to peak L-dopa plasma concentrations. Mean on time was 21.9% (37 min) longer with 30 g mucuna than with LD/CD (p = 0.021); peak L-dopa plasma concentrations were 110% higher and the area under the plasma concentration v time curve (area under curve) was 165.3% larger (p = 0.012). No significant differences in dyskinesias or tolerability occurred. CONCLUSIONS: The rapid onset of action and longer on time without concomitant increase in dyskinesias on mucuna seed powder formulation suggest that this natural source of L-dopa might possess advantages over conventional L-dopa preparations in the long term management of PD. Assessment of long term efficacy and tolerability in a randomised, controlled study is warranted. PMID- 15548482 TI - Repetitive training of compensatory steps: a therapeutic approach for postural instability in Parkinson's disease. AB - BACKGROUND: Postural instability (PI) is a common and serious problem in Parkinson's disease (PD). Dopaminergic medication is of negligible use and a positive effect of deep brain stimulation on this issue has not been reported. OBJECTIVE: To develop a method of repetitive training of compensatory steps to enhance protective postural responses by using training strategies based on recent neurophysiological research. METHODS: Fourteen patients with PD took part in a multiple baseline design study and were trained for 14 days in an ambulant setting consisting of two daily sessions. RESULTS: After training, the length of compensatory steps increased and the step initiation shortened. In a gait analysis, the cadence and the step length increased, gait velocity improved, and the period of double support shortened. The "mobility" subscore of a quality of life questionnaire (PDQ-39) also improved. All these changes were significant (p < 0.05). These effects were stable for two months without additional training. CONCLUSION: The repetitive training of compensatory steps is an effective approach in the treatment of PI and should be applied if PI is evident clinically or in a patient's history. PMID- 15548483 TI - Factors predicting improvement in motor disability in writer's cramp treated with botulinum toxin. AB - OBJECTIVE: To identify factors predicting improvement in motor disability in writer's cramp treated with botulinum toxin (BTX). METHODS: 47 patients with writer's cramp were treated with BTX and were evaluated by the same neurologists at initial referral, after each BTX injection, and when the effect of BTX was maximal at the time of the study. Patients and examiners simultaneously and independently rated the efficacy of BTX injections. Self assessment was a global clinical impression of the impact of treatment on writing quality, writing speed, writing errors, and legibility of handwriting; for objective assessment, the examiners used the Burke-Fahn-Marsden (BFM) scale. RESULTS: On the BFM scale, there was a significant improvement (p<0.0001) in both severity and disability scores. Patients with a pronation/flexion pattern of dystonia showed the best and the most sustained improvement. Primary writing tremor was little improved. There was a correlation between the self assessment score and the Burke-Fahn-Marsden score. Benefit was maintained over time CONCLUSIONS: These results have implications for the identification of patients most likely to benefit from BTX injections. PMID- 15548484 TI - Apolipoprotein E genotypes do not influence the age of onset in Huntington's disease. AB - OBJECTIVE: The epsilon4 allele of the apolipoprotein E (ApoE) gene has been defined as a critical factor for early onset neurodegeneration in Pick's, Parkinson's, and Alzheimer's disease. Unexpectedly, the epsilon4 allele appeared to delay the age of onset in Huntington's disease (HD) patients. Furthermore, sex specific effects were reported on earlier age of onset due to the ApoE epsilon2epsilon3 genotype in males with HD. The age of onset of HD is known to be negatively correlated with increasing lengths of pathogenetic CAG expansions in the huntingtin gene. METHODS: In order to examine the effects of CAG block lengths, we have correlated ApoE genotypes with the age of onset in 145 patients symptomatic for HD with psychiatric and somatic symptoms (depression, psychosis, dementia, choreic, and other movement disorders) harbouring only modestly expanded huntingtin alleles (41-45 CAGs). RESULTS: The negative correlation between age of onset and CAG block length was established in our HD cohort. Statistically significant effects of the epsilon4 allele were not obvious regarding clinical characteristics including age of onset, nor were any sex differences for the epsilon2epsilon3 genotype observed. CONCLUSION: The ApoE genotype does not affect the course of HD significantly. PMID- 15548485 TI - Pattern of collaterals, type of infarcts, and haemodynamic impairment in carotid artery occlusion. AB - BACKGROUND: In internal carotid artery (ICA) occlusion, increased oxygen extraction fraction (OEF) indicates inadequate collateral blood flow distal to the occlusion, which may be caused by poor function of collateral pathways. In ICA occlusion, the circle of Willis may be the major collateral pathway, while the collaterals through the ophthalmic artery and leptomeningeal vessels may be recruited when collateral flow through the circle of Willis is inadequate. Conversely, ischaemic lesions may affect the adequacy of collateral blood flow by reducing the metabolic demand of the brain. OBJECTIVE: To determine whether the pattern of collateral pathways and the type of infarcts are independent predictors of OEF in ICA occlusion. METHODS: We studied 42 patients with symptomatic ICA occlusion. The presence of Willisian, ophthalmic, or leptomeningeal collaterals was evaluated by conventional four vessel angiography. The infarcts on magnetic resonance imaging were categorised as territorial, border zone (external or internal), striatocapsular, lacunar, and other white matter infarcts. The value of OEF in the affected hemisphere was measured with positron emission tomography as an index of haemodynamic impairment. RESULTS: Using multivariate analysis, the presence of any ophthalmic or leptomeningeal collaterals and the absence of striatocapsular infarcts were significant and independent predictors of increased OEF. CONCLUSIONS: In patients with symptomatic ICA occlusion, the supply of collateral flow, which is affected by the pattern of collateral pathways, and the metabolic demand of the brain, which is affected by the type of infarct, may be important factors determining the severity of haemodynamic impairment. PMID- 15548486 TI - Variations in care and outcome in the first year after stroke: a Western and Central European perspective. AB - BACKGROUND: There are significant variations in the short term patterns of care and outcome after a first stroke in Europe. OBJECTIVE: To estimate the variation in stroke care and outcome up to 1 year after a stroke in selected European centres. METHODS: Hospital based stroke registers were established in 11 hospitals in seven western and central European countries to collect demographic, clinical, and resource use data at the time of first ever stroke during 1993-4. At 3 and 12 months, details of survival, activities of daily living score, and use of services were recorded. Univariate comparisons between centres were made using the chi2 test and stepwise regression was used to identify associations between centre, case mix, therapy provision, and outcomes. RESULTS: Of the 4048 patients registered, 23% were lost to follow up and 38% had died at 1 year. The proportions of survivors who felt they needed assistance at 12 months ranged from 35% in Italy to 77% in UK2. There were comparatively high amounts of therapy provided up to 1 year in UK3, France and Germany 1, mainly at home. At 1 year, social services were still providing support in UK1, UK5 and France, with some support in Germany 1 and family support was provided in France. In multivariate analysis, after adjustment for case mix and receipt of rehabilitation, non-UK centres had improved activities of daily living (p<0.001). Older age was indicative of more need for assistance, but less likelihood of assistance from the family. Those in France were more likely to get assistance from their family than any other centre. Mainland European patients were more likely to get help from their family than those in the UK. Patients in all areas except UK2 and UK3 were more likely to be dead or dependent at 1 year than patients in UK1. CONCLUSIONS: There were significant variations in the pathways of care for stroke across European centres in the mid 1990s, which were associated with variation in outcome, and remain unexplained. Family support is more prevalent in southern Europe and service support more prevalent in the UK. PMID- 15548487 TI - Circumscribed unilateral lower pontine lesion. PMID- 15548488 TI - Personality change after stroke: some preliminary observations. AB - OBJECTIVES: To describe changes in personality after stroke and effects on carers. METHODS: A consecutive series of patients was recruited from hospital admissions with stroke. A novel questionnaire was administered to the patients' main carer at nine months after the stroke to determine their perception of the patients' pre-stroke and post-stroke personality. Personality change was identified by changes in these ratings, and associations between personality change and the following variables explored: emotional disorder in patients and carers (measured using the hospital anxiety and depression scale and a structured psychiatric interview), stroke classification (Oxford community stroke classification), residual disability (Barthel index and Nottingham extended activities of daily living scale), and lesion characteristics on computed tomography (CT). RESULTS: Carers of 35 patients with stroke took part. Reported changes in personality after stroke included: reduced patience and increased frustration (both p<0.0001, t test of difference), reduced confidence, more dissatisfaction, and a less easy going nature (all p<0.005). Occasionally, aspects of personality change were seen as positive by carers. There were relations between greater personality change and interviewer rated patient depression or anxiety (p<0.001) but not when this was self rated; and between personality change and both emotional disorder in carers (p<0.005) and greater disability (p<0.01) but not CT lesion characteristics. CONCLUSIONS: Carers commonly perceive personality change in stroke patients. This is associated with self rated emotional distress in the carer. More research is needed to understand what carers mean by "personality change" and what factors contribute to the perceived change. PMID- 15548489 TI - Temporal evolution of water diffusion parameters is different in grey and white matter in human ischaemic stroke. AB - OBJECTIVES: Our purpose was to investigate whether differences exist in the values and temporal evolution of mean diffusivity () and fractional anisotropy (FA) of grey and white matter after human ischaemic stroke. METHODS: Thirty two patients with lesions affecting both grey and white matter underwent serial diffusion tensor magnetic resonance imaging (DT-MRI) within 24 hours, and at 4-7 days, 10-14 days, 1 month, and 3 months after stroke. Multiple small circular regions of interest (ROI) were placed in the grey and white matter within the lesion and in the contralateral hemisphere. Values of [grey], [white], FA[grey] and FA[white] were measured in these ROI at each time point and the ratios of ischaemic to normal contralateral values (R and FAR) calculated. RESULTS: and FA showed different patterns of evolution after stroke. After an initial decline, the rate of increase of [grey] was faster than [white] from 4-7 to 10-14 days. FA[white] decreased more rapidly than FA[grey] during the first week, thereafter for both tissue types the FA decreased gradually. However, FA[white] was still higher than FA[grey] at three months indicating that some organised axonal structure remained. This effect was more marked in some patients than in others. R[grey] was significantly higher than R[white] within 24 hours and at 10-14 days (p<0.05), and FAR[white] was significantly more reduced than FAR[grey] at all time points (p<0.001). CONCLUSIONS: The values and temporal evolution of and FA are different for grey and white matter after human ischaemic stroke. The observation that there is patient-to-patient variability in the degree of white matter structure remaining within the infarct at three months may have implications for predicting patient outcome. PMID- 15548490 TI - Deficient saccadic inhibition in Asperger's disorder and the social-emotional processing disorder. AB - BACKGROUND: Both Asperger's disorder and the social-emotional processing disorder (SEPD), a form of non-verbal learning disability, are associated with executive function deficits. SEPD has been shown to be associated with deficient saccadic inhibition. OBJECTIVE: To study two executive functions in Asperger's disorder and SEPD, inhibition and task switching, using a single saccadic paradigm. METHODS: 22 control subjects and 27 subjects with developmental social processing disorders-SEPD, Asperger's disorder, or both syndromes-performed random sequences of prosaccades and antisaccades. This design resulted in four trial types, prosaccades and antisaccades, that were either repeated or switched. The design allowed the performance costs of inhibition and task switching to be isolated. RESULTS: Subjects with both Asperger's disorder and SEPD showed deficient inhibition, as indicated by increased antisaccade errors and a disproportionate increase in latency for antisaccades relative to prosaccades. In contrast, task switching error and latency costs were normal and unrelated to the costs of inhibition. CONCLUSIONS: This study replicates the finding of deficient saccadic inhibition in SEPD, extends it to Asperger's disorder, and implicates prefrontal cortex dysfunction in these syndromes. The finding of intact task switching shows that executive function deficits in Asperger's disorder and SEPD are selective and suggests that inhibition and task switching are mediated by distinct neural networks. PMID- 15548491 TI - Neurosyphilis in the modern era. AB - OBJECTIVE: To review the nature of the presentation of neurosyphilis, the value of diagnostic tests, and the classification of the disease. METHODS: A retrospective review was carried out of the records of patients who had been identified as possible cases of neurosyphilis by a positive FTA-abs test in the CSF. The review extended over 10 years at a single hospital which served a population of mixed ancestry in a defined catchment area in the Western Cape province of South Africa. Patients were placed in predefined diagnostic categories, and clinical, radiological, and laboratory features were assessed. RESULTS: 161 patients met diagnostic criteria for neurosyphilis: 82 presented with combinations of delirium and dementia and other neuropsychiatric conditions, and the remainder had typical presentations such as stroke (24), spinal cord disease (15), and seizures (14). The average age of presentation ranged from 35.9 to 42.6 years in the different categories of neurosyphilis. Of those followed up, 77% had residual deficits from their initial illness. Cerebrospinal fluid (CSF) VDRL was positive in 73% of cases. CONCLUSIONS: The diagnosis of neurosyphilis can be made with reasonable certainty if there is an appropriate neuropsychiatric syndrome associated with a positive CSF VDRL. If the VDRL is negative, a positive FTA-abs in an appropriate clinical setting, associated with raised CSF cell count, protein, or IgG index, is a useful method of identifying neurosyphilis. Tabes dorsalis has become uncommon, but this is likely to be the only manifestation of neurosyphilis that has been altered during the antibiotic era. PMID- 15548492 TI - Cell response to oxidative stress induced apoptosis in patients with Leber's hereditary optic neuropathy. AB - OBJECTIVES: Leber's hereditary optic neuropathy (LHON) is a maternally inherited disease in which acute or subacute bilateral visual loss occurs preferentially in young men. Over 95% of LHON cases are associated with one of three mitochondrial DNA (mtDNA) point mutations, but only 50% of men and 10% of women who harbour a pathogenetic mtDNA mutation develop optic neuropathy. This incomplete penetrance and preference for men suggests that additional genetic (nuclear or mitochondrial) and/or environmental factors must modulate phenotype expression in LHON. A role for reactive oxygen species (ROS) in mitochondrial diseases, secondary to mtDNA mutations, or as a result of the direct effect of ROS cytotoxicity, has been implicated in many mitochondrial disorders, including LHON. The purpose of this study was to investigate the role of oxidative stress induced apoptosis in LHON. METHODS: The 2-deoxy-D-ribose induced apoptotic response of peripheral blood lymphocytes from six patients with LHON and six healthy subjects was investigated using light microscopy, flow cytometry, agarose gel electrophoresis, and the measurement of mitochondrial membrane potential. RESULTS: Cells of patients with LHON had a higher rate of apoptosis than those of controls and there was evidence of mitochondrial involvement in the activation of the apoptotic cascade. CONCLUSIONS: These differences in oxidative stress induced apoptosis are in line with the hypothesis that redox homeostasis could play a role in the expression of genetic mutations in different individuals and could represent a potential target in the development of new therapeutic strategies. PMID- 15548493 TI - The effects of water ingestion on orthostatic hypotension in two groups of chronic autonomic failure: multiple system atrophy and pure autonomic failure. AB - BACKGROUND: Oral ingestion of water increases seated blood pressure in patients with chronic autonomic failure by mechanisms that remain unclear. As orthostatic hypotension is common in chronic autonomic failure, and is not always adequately controlled by medication, the potential benefits of water ingestion on standing blood pressure were studied in two types of autonomic failure: multiple system atrophy (MSA), in which the lesion is central and pre-ganglionic, and pure autonomic failure (PAF), in which the lesion is post-ganglionic. METHODS: In 14 patients with autonomic failure (seven PAF and seven MSA) standing blood pressure and heart rate were measured before, and 15 and 35 minutes after ingestion of 480 ml distilled water. Patients remained seated for 15 minutes after water ingestion, with beat to beat cardiovascular indices measured with the Portapres II device with subsequent Modelflow analysis. RESULTS: Standing prior to water ingestion caused a significant fall in blood pressure in all patients. After water ingestion there was a rise in seated blood pressure. Seated and standing blood pressure at 15 and 35 minutes after water ingestion was significantly higher than before water, with an improvement in orthostatic symptoms. The time to first significant rise in seated blood pressure occurred at 5 minutes post water ingestion in PAF and at 13 minutes in MSA. These increases were accompanied by increases in total peripheral resistance, reaching significance by 5 minutes in PAF and 13 minutes in MSA. There were no significant changes in cardiac output, stroke volume, or ejection fraction. CONCLUSIONS: Water is thus beneficial in improving standing BP in AF, acting within 15 minutes in both MSA and PAF. The earlier onset of the pressor effect in PAF may reflect the differing lesion site and underlying pathophysiology between these conditions. PMID- 15548494 TI - Samuel Hahnemann's doctoral dissertation of 1779. PMID- 15548495 TI - Similar early clinical presentations in familial and non-familial frontotemporal dementia. AB - BACKGROUND: It is unclear whether there are early clinical features that can distinguish between patients with familial and non-familial frontotemporal dementia (FTD). OBJECTIVE: To compare the clinical features of FTD cases who have tau gene mutations with those of cases with a family history of FTD but no tau gene mutation, and with sporadic cases with neither feature. METHODS AND RESULTS: Comparisons of the behavioural, cognitive, and motor features in 32 FTD patients (five positive for tau gene mutations, nine familial but tau negative, and 18 tau negative sporadic) showed that age of onset and duration to diagnosis did not differ between the groups. Apathy was not observed in tau mutation positive cases, and dysexecutive signs were more frequent in familial tau mutation negative cases. Memory deficits and behavioural changes were common in all groups. CONCLUSIONS: In comparison with other neurodegenerative conditions such as Alzheimer's disease and Parkinson's disease, neither tau gene mutations nor strong familial associations confer earlier disease susceptibility. PMID- 15548496 TI - Polymorphisms in neprilysin gene affect the risk of Alzheimer's disease in Finnish patients. AB - OBJECTIVES: Neprilysin (NEP) is an amyloid beta-peptide (Abeta) degrading enzyme expressed in the brain, and accumulation of Abeta is the neuropathological hallmark in Alzheimer's disease (AD). In this study we investigated whether polymorphisms in the NEP gene have an effect on the risk for AD. METHODS: The frequencies of seven single nucleotide polymorphisms (SNPs) and apolipoprotein E (APOE) were assessed in 390 AD patients and 468 cognitively healthy controls. Genotypes of the study groups were compared using binary logistic regression analysis. Haplotype frequencies of the SNPs were estimated from genotype data. RESULTS: Two SNPs, rs989692 and rs3736187, had significantly different allelic and genotypic frequencies (uncorrected p = 0.01) between the AD and the control subjects and haplotype analysis showed significant association between AD and NEP polymorphisms. CONCLUSION: Taken together, these findings suggest that polymorphisms in the NEP gene increase risk for AD and support a potential role for NEP in AD. PMID- 15548497 TI - Olfactory function distinguishes vascular parkinsonism from Parkinson's disease. AB - OBJECTIVE: To compare olfactory function in vascular parkinsonism and Parkinson's disease diagnosed according to published clinical diagnostic criteria. METHODS: The University of Pennsylvania smell identification test (UPSIT) was carried out in 14 patients with vascular parkinsonism, 18 with Parkinson's disease, and 27 normal controls matched for age, sex, and smoking status. RESULTS: UPSIT scores in vascular parkinsonism (mean 26.1, 95% confidence interval, 23.1 to 29.0) were significantly better than in Parkinson's disease (mean 17.1 (14.5 to 19.7)) (p<0.0001), and did not differ from the healthy controls (mean 27.6 (25.8 to 29.4)) (p = 0.32). CONCLUSIONS: Testing olfactory function may be helpful in differentiating vascular parkinsonism from Parkinson's disease. PMID- 15548498 TI - Unexpected decline in survival from amyotrophic lateral sclerosis/motor neurone disease. AB - OBJECTIVES: To describe survival of 1226 Scottish adults with amyotrophic lateral sclerosis/motor neurone disease (ALS/MND). METHODS: Ten year, prospective, population based disease register. Cox time dependent proportional hazards modelling for multivariate survival analyses. RESULTS: Median survival from onset was 25 months (interquartile range 16-34 months). In multivariate models we found an increased hazard with more recently diagnosed cases-that is, there was an unexpected decline in survival over the 10 year period (hazard ratio (HR) 1.06 (95% CI 1.04 to 1.09). Positive effects on survival were demonstrated for longer time from onset to diagnosis (HR 0.38 (95% CI 0.33 to 0.42), assessment by a neurological specialist (HR 0.56 (95% CI 0.40 to 0.77), and treatment with riluzole (HR 0.24 (95% CI 0.14 to 0.42). Poor prognosis was associated with bulbar onset (HR 1.25 (95% CI 1.09 to 1.46) and a mixed lower and upper motor neurone syndrome (HR 1.23 (95% CI 1.01-1.49) and increasing age. CONCLUSIONS: We found an unexpected decline in survival over the 10 year period, despite controlling for potential confounding variables. We would be cautious about over interpreting these observations and suggest that further research is required to confirm or refute these findings. PMID- 15548499 TI - Predictive value of death certification in the case ascertainment of epilepsy. AB - OBJECTIVES: Death certificates are an unreliable source of information on cause of death, and mortality due to epilepsy can thus be underestimated. METHODS: We investigated people with epilepsy who had died, and attempted to identify factors that influence inclusion of epilepsy on the death certificate; eight factors were hypothesised and entered into a univariate logistic regression model. RESULTS: Epilepsy was on the death certificate of 16/243 (7%) people who had had epilepsy. Factors that influenced whether or not epilepsy appeared on the certificate were seizure frequency, antiepileptic drug treatment, cause of death, and certifying physician. Factors that did not seem to influence the inclusion of epilepsy were presence of convulsive seizures, occurrence of seizures during follow up, and age at death. CONCLUSIONS: We have estimated the degree of unreliability of death certificates (as currently used in the UK) as a source of information on cause of death in epilepsy. We have found that epilepsy may not appear on death certificates even if people had active epilepsy. PMID- 15548500 TI - Effect of body size on operative risk of carotid endarterectomy. AB - Many studies have found that women have a higher risk of perioperative stroke or death from carotid endarterectomy. Other vascular surgical procedures have demonstrated that body size and morphology impact on operative risk. We correlated the 30 day operative risk of stroke and death in the European Carotid Surgery Trial (ECST) with height, weight, body surface area (BSA), and body mass index using single variable analyses and multivariable logistic regression. Women were at significantly higher risk of perioperative stroke and death in the ECST. Both height and BSA confounded the effect of sex, implying that the generally smaller size of women may contribute to their increased risk. This finding should be validated in other large datasets. PMID- 15548501 TI - Phenotypic variability in siblings with type III spinal muscular atrophy. AB - Autosomal recessive spinal muscular atrophy (SMA) shows substantial phenotypic variability, presenting at a variety of ages from infancy to adult life. Diagnostic difficulties may arise because SMA sometimes produces a dystrophic or myopathic phenotype rather than classical neurogenic abnormalities. Two brothers are described who illustrate this principle and highlight the increasing importance of molecular genetics in investigating patients with neuromuscular diseases. The findings are discussed in the light of recent observations in a mouse model of SMA. PMID- 15548502 TI - Antiganglioside antibodies in paraneoplastic peripheral neuropathies. AB - A total of 29 patients with cancer and neuropathies of unknown origin that were possibly paraneoplastic were tested for antiganglioside antibodies by immunodot blot and ELISA. None of the patients had onconeural antibodies. They were compared with 41 normal subjects and 187 patients with metabolic or idiopathic neuropathies. Antiganglioside antibodies, mainly IgM anti-GM1, were more frequently found in the patients with cancer than in the control groups. However, the levels of antibodies were not different from those of the controls. There was no correlation with the pattern of the neuropathy. These results do not support the hypothesis that antiganglioside antibodies are frequent and major immunological targets in paraneoplastic neuropathies. PMID- 15548503 TI - Primary lateral sclerosis presenting parkinsonian symptoms without nigrostriatal involvement. AB - We encountered three patients with primary lateral sclerosis (PLS) showing bradykinesia, frozen gait, and severe postural instability, as well as slowly progressive spinobulbar spasticity. Cranial magnetic resonance (MR) imaging showed precentral gyrus atrophy. Central motor conduction was markedly prolonged or failed to evoke a response. Positron emission tomography (PET) showed significant reduction of [18F]fluoro-2-deoxy-D-glucose uptake in the area of the precentral gyrus extending to the prefrontal, medial frontal, and cingulate areas. No abnormalities were seen in the nigrostriatal system with PET using [18F]fluorodopa or [11C]raclopride or with proton MR spectroscopy. Thus, widespread prefrontal, medial, and cingulate frontal lobe involvement can be associated with the parkinsonian symptoms in PLS. PMID- 15548504 TI - Pathological substrate for regional distribution of increased atrophy rates in progressive supranuclear palsy. AB - BACKGROUND: Most magnetic resonance imaging (MRI) studies of progressive supranuclear palsy (PSP) are cross-sectional and lack post mortem confirmation of the diagnosis. MRI features described previously in PSP correspond to regions of pathological involvement demonstrated in separate studies, but serial MRI with pathological follow up has not been undertaken. OBJECTIVE: To investigate whether regions of increased atrophy rates demonstrated in PSP during life using fluid registered serial MRI correspond with pathological findings in confirmed PSP. METHODS: A 59 year old male presented with a six month history of balance problems and dysarthria. He had a symmetrical, levodopa unresponsive akinetic rigid syndrome with a vertical supranuclear gaze palsy. A clinical diagnosis of probable PSP was made. His disease progressed relentlessly and he died five years after onset. Two serial MRI scans undertaken during life were reviewed and fluid (non-linear) registration of the images carried out. Post mortem histopathological analysis of the brain was undertaken to definitively confirm the diagnosis and compare regional pathology with the serial imaging. RESULTS: Fluid registration demonstrated greatest rates of atrophy in the brainstem and frontal cortex, in keeping with the distribution of pathology seen at autopsy. CONCLUSION: Fluid registration of serial MRI allows the topography and rates of regional atrophy in PSP to be delineated in life. Atrophy patterns correlated well with regional pathological load. These observations suggest that serial MRI with registration may help differentiate PSP from clinically similar conditions and supports its use as a surrogate marker of disease progression. PMID- 15548505 TI - Patients whom neurologists find difficult to help. AB - BACKGROUND: All doctors recognise that some patients are more "difficult to help" than others, but the issue has received little systematic investigation in neurological practice. OBJECTIVE: To test the hypothesis that patients whose symptoms were less explained by organic disease would be perceived as more difficult to help. METHODS: In a consecutive series of 300 new neurology outpatients, neurologists indicated on four point Likert-type scales how "difficult to help" they found the patient and to what extent the patient's symptoms were explained by organic disease. The patients' demographics, health status, number of somatic symptoms, and mental state were also assessed. RESULTS: The neurologists rated 143 patients (48%) as "not at all difficult" to help, 111 (37%) as "somewhat difficult", 27 (9%) as "very difficult", and 18 (6%) as "extremely difficult". A logistic regression model was constructed and the hypothesis that patients whose symptoms were less explained by organic disease would be perceived as more difficult to help was supported. The only other measured variable that contributed to perceived difficulty was physical disability, but it explained only a small amount of the variance. CONCLUSIONS: Neurologists find patients whose symptoms are not explained by organic disease more difficult to help than their other patients. PMID- 15548506 TI - Tissue at risk concept for endovascular treatment of severe vasospasm after aneurysmal subarachnoid haemorrhage. AB - OBJECTIVE: To report a case of severe vasospasm after subarachnoid haemorrhage (SAH) where "tissue at risk" was identified by magnetic resonance imaging (MRI), and to demonstrate the haemodynamic consequences with either resolution of the perfusion-diffusion mismatch by balloon angioplasty or evolution of an infarct. METHODS: A 45 year old women with SAH underwent surgical treatment of a ruptured middle cerebral artery (MCA) aneurysm. On day 3 she became obtunded and developed a right hemiparesis. Diffusion weighted (DWI) and perfusion weighted (PWI) imaging were done before and after transluminal balloon angioplasty (TBA) of multifocal proximal vasospasm. RESULTS: The initial MRI revealed no DWI lesion but PWI showed a severe perfusion deficit of 6.7 to 16.4 seconds in the complete left MCA territory. Digital subtraction angiography confirmed severe segmental narrowing of left C1 and M1. The spastic segments were successfully dilated by TBA. Follow up MRI showed that the PWI-DWI mismatch resolved in the anterior and middle MCA territory with no tissue infarction, whereas in the terminal dorsal MCA territory a severe mismatch remained and cerebral infarction evolved. CONCLUSIONS: PWI/DWI can identify tissue at risk for infarction in severe vasospasm following SAH. This may allow selection of patients for angioplasty and the monitoring of treatment effects. PMID- 15548507 TI - Delayed early morning turn "ON" in response to a single dose of levodopa in advanced Parkinson's disease: pharmacokinetics should be considered. PMID- 15548508 TI - Lethal encephalopathy in a patient with isolated nervous system vasculitis. PMID- 15548509 TI - Acute combined central and peripheral inflammatory demyelination. PMID- 15548510 TI - Miller Fisher syndrome associated with Pasteurella multocida infection. PMID- 15548511 TI - Coagulopathy and NICE recommendations for patients with mild head injury. PMID- 15548512 TI - Neurogenic T wave inversion in pure left insular stroke associated with hyperhomocysteinaemia. PMID- 15548513 TI - Prion disease at a regional neuroscience centre: retrospective audit. PMID- 15548514 TI - The tight junction protein occludin and the adherens junction protein alpha catenin share a common interaction mechanism with ZO-1. AB - The exact sites, structures, and molecular mechanisms of interaction between junction organizing zona occludence protein 1 (ZO-1) and the tight junction protein occludin or the adherens junction protein alpha-catenin are unknown. Binding studies by surface plasmon resonance spectroscopy and peptide mapping combined with comparative modeling utilizing crystal structures led for the first time to a molecular model revealing the binding of both occludin and alpha catenin to the same binding site in ZO-1. Our data support a concept that ZO-1 successively associates with alpha-catenin at the adherens junction and occludin at the tight junction. Strong spatial evidence indicates that the occludin C terminal coiled-coil domain dimerizes and interacts finally as a four-helix bundle with the identified structural motifs in ZO-1. The helix bundle of occludin406-521 and alpha-catenin509-906 interacts with the hinge region (ZO-1591 632 and ZO-1591-622, respectively) and with (ZO-1726-754 and ZO-1756-781) in the GuK domain of ZO-1 containing coiled-coil and alpha-helical structures, respectively. The selectivity of both protein-protein interactions is defined by complementary shapes and charges between the participating epitopes. In conclusion, a common molecular mechanism of forming an intermolecular helical bundle between the hinge region/GuK domain of ZO-1 and alpha-catenin and occludin is identified as a general molecular principle organizing the association of ZO-1 at adherens and tight junctions. PMID- 15548515 TI - Structure of a high fidelity DNA polymerase bound to a benzo[a]pyrene adduct that blocks replication. AB - Of the carcinogens to which humans are most frequently exposed, the polycyclic aromatic hydrocarbon benzo[a]pyrene (BP) is one of the most ubiquitous. BP is a byproduct of grilled foods and tobacco and fuel combustion and has long been linked to various human cancers, particularly lung and skin. BP is metabolized to diol epoxides that covalently modify DNA bases to form bulky adducts that block DNA synthesis by replicative or high fidelity DNA polymerases. Here we present the structure of a high fidelity polymerase from a thermostable strain of Bacillus stearothermophilus (Bacillus fragment) bound to the most common BP derived N2-guanine adduct base-paired with cytosine. The BP adduct adopts a conformation that places the polycyclic BP moiety in the nascent DNA minor groove and is the first structure of a minor groove adduct bound to a polymerase. Orientation of the BP moiety into the nascent DNA minor groove results in extensive disruption to the interactions between the adducted DNA duplex and the polymerase. The disruptions revealed by the structure of Bacillus fragment bound to a BP adduct provide a molecular basis for rationalizing the potent blocking effect on replication exerted by BP adducts. PMID- 15548516 TI - The assembly of MreB, a prokaryotic homolog of actin. AB - MreB, a major component of the bacterial cytoskeleton, exhibits high structural homology to its eukaryotic counterpart actin. Live cell microscopy studies suggest that MreB molecules organize into large filamentous spirals that support the cell membrane and play a key shape-determining function. However, the basic properties of MreB filament assembly remain unknown. Here, we studied the assembly of Thermotoga maritima MreB triggered by ATP in vitro and compared it to the well-studied assembly of actin. These studies show that MreB filament ultrastructure and polymerization depend crucially on temperature as well as the ions present on solution. At the optimal growth temperature of T. maritima, MreB assembly proceeded much faster than that of actin, without nucleation (or nucleation is highly favorable and fast) and with little or no contribution from filament end-to-end annealing. MreB exhibited rates of ATP hydrolysis and phosphate release similar to that of F-actin, however, with a critical concentration of approximately 3 nm, which is approximately 100-fold lower than that of actin. Furthermore, MreB assembled into filamentous bundles that have the ability to spontaneously form ring-like structures without auxiliary proteins. These findings suggest that despite high structural homology, MreB and actin display significantly different assembly properties. PMID- 15548517 TI - Adipocytic differentiation and liver x receptor pathways regulate the accumulation of triacylglycerols in human vascular smooth muscle cells. AB - Lipid accumulation by vascular smooth muscle cells (VSMC) is a feature of atherosclerotic plaques. In this study we describe two mechanisms whereby human VSMC foam cell formation is driven by de novo synthesis of fatty acids leading to triacylglycerol accumulation in intracellular vacuoles, a process distinct from serum lipoprotein uptake. VSMC cultured in adipogenic differentiation medium accumulated lipids and were induced to express the adipocyte marker genes adipsin, adipocyte fatty acid-binding protein, C/EBPalpha, PPARgamma, and leptin. However, complete adipocyte differentiation was not observed as numerous genes present in mature adipocytes were not detected, and the phenotype was reversible. The rate of lipid accumulation was not affected by PPARgamma agonists, but screening for the effects of other nuclear receptor agonists showed that activation of the liver X receptors (LXR) dramatically promoted lipid accumulation in VSMC. Both LXRalpha and LXRbeta were present in VSMC, and their activation with TO901317 resulted in induction of the lipogenic genes fatty acid synthetase, sterol regulatory element binding protein (SREBP1c), and stearoyl-CoA desaturase. 27-Hydroxycholesterol, an abundant oxysterol synthesized by VSMC acted as an LXR antagonist and, therefore, may have a protective role in preventing foam cell formation. Immunohistochemistry showed that VSMC within atherosclerotic plaques express adipogenic and lipogenic markers, suggesting these pathways are present in vivo. Moreover, the development of an adipogenic phenotype in VSMC is consistent with their known phenotypic plasticity and may contribute to their dysfunction in atherosclerotic plaques and, thus, impinge on plaque growth and stability. PMID- 15548518 TI - Loss of net as repressor leads to constitutive increased c-fos transcription in cervical cancer cells. AB - We have investigated the expression of c-fos in cervical carcinoma cells and in somatic cell hybrids derived therefrom. In malignant cells, c-fos was constitutively expressed even after serum starvation. Dissection of the c-fos promoter showed that expression was mainly controlled by the SRE motif, which was active in malignant cells, but repressed in their non-malignant counterparts. Constitutive SRE activity was not mediated by sustained mitogen-activated protein kinase activity but because of inefficient expression of the ternary complex factor Net, which was either very low or even barely discernible. Chromatin immunoprecipitation assays revealed that Net directly binds to the SRE nucleoprotein complex in non-tumorigenic cells, but not in malignant segregants. Small interfering RNA targeted against Net resulted in enhanced c-fos transcription, clearly illustrating its repressor function. Conversely, stable ectopic expression of Net in malignant cells negatively regulated endogenous c fos, resulting in a disappearance of the c-Fos protein from the AP-1 transcription complex. These data indicate that loss of Net and constitutive c fos expression appear to be a key event in the transformation of cervical cancer cells. PMID- 15548519 TI - STAT-3-dependent cytosolic phospholipase A2 expression is required for thrombin induced vascular smooth muscle cell motility. AB - Vascular smooth muscle cell (VSMC) migration from media to intima and its multiplication in intima is a contributing factor in the pathogenesis of atherosclerosis and restenosis after angioplasty. Previously, we have demonstrated that STAT-3-dependent cytosolic phospholipase A(2) (cPLA(2)) expression is needed for VSMC motility induced by platelet-derived growth factor BB, a receptor tyrosine kinase agonist (Neeli et al. (2005) J. Biol. Chem. 279, 46122-46128). In order to learn more about the STAT-3-cPLA(2) axis in motogenic signaling, here we have studied its role in VSMC motility in response to a G protein-coupled receptor (GPCR) agonist, thrombin. Thrombin induced VSMC motility in a dose-dependent manner with a maximum effect at 0.5 units/ml. Thrombin activated STAT-3 as measured by its tyrosine phosphorylation and translocation from the cytoplasm to the nucleus. Forced expression of a dominant negative mutant of STAT-3 reduced thrombin-induced STAT-3 tyrosine phosphorylation and its translocation from the cytoplasm to the nucleus. Thrombin stimulated STAT-3-DNA binding and reporter gene activities in VSMC, and these responses were blocked by FS3DM, a dominant negative mutant of STAT-3. FS3DM also attenuated thrombin induced VSMC motility. Thrombin induced the expression of cPLA(2) in a time- and STAT-3-dependent manner. In addition, pharmacological inhibition of cPLA(2) blocked thrombin-induced VSMC motility. Furthermore, exogenous addition of arachidonic acid rescued thrombin-induced VSMC motility from inhibition by blockade of STAT-3 activation. Forced expression of cPLA(2) also surpassed the inhibitory effect of dominant negative STAT-3 on thrombin-induced VSMC motility. Together, these results show that thrombin-induced VSMC motility requires STAT-3 dependent induction of expression of cPLA(2). PMID- 15548520 TI - Tumor suppressor APC blocks DNA polymerase beta-dependent strand displacement synthesis during long patch but not short patch base excision repair and increases sensitivity to methylmethane sulfonate. AB - In the present investigation, we report a previously unsuspected function of the tumor suppressor protein, APC (adenomatous polyposis coli), in the regulation of base excision repair (BER). We identified a proliferating cell nuclear antigen interacting protein-like box sequence in APC that binds DNA polymerase beta and blocks DNA polymerase beta-mediated strand-displacement synthesis in long patch BER without affecting short patch BER. We further showed that the colon cancer cell line expressing the wild-type APC gene was more sensitive to a DNA methylating agent due to decreased DNA repair by long patch BER than the cell line expressing the mutant APC gene lacking the proliferating cell nuclear antigen-interacting protein-like box. Experiments based on RNA interference showed that the wild-type APC gene expression is required for DNA methylation induced sensitivity of colon cancer cells. Thus, APC may play a critical role in determining utilization of long versus short patch BER pathways and affect the susceptibility of colon cancer cells to carcinogenic and chemotherapeutic agents. PMID- 15548521 TI - The CSB protein actively wraps DNA. AB - The CSB protein is a member of the SWI2/SNF2 family of ATP-dependent chromatin remodeling factors and is essential for transcription-coupled DNA repair. The role of CSB in this DNA repair process is unclear, but the protein was found to remodel nucleosomes and alter DNA double helix conformation upon binding. Elucidating the nature of the change in DNA structure induced by CSB is of great interest for understanding the CSB mechanism of action. We analyzed the CSB.DNA complex by scanning force microscopy and measured a shortening of DNA contour length upon CSB binding in the presence of ATP. This DNA length reduction most likely results from DNA wrapping around the protein. Shorter DNA molecules were observed more frequently in the presence of non-hydrolyzable ATP analogues. These results suggest that DNA wrapping depends on ATP binding, whereas ATP hydrolysis results in unwrapping. We also provide evidence suggesting that CSB binds DNA as a dimer. DNA wrapping and unwrapping allows CSB to actively alter the DNA double helix conformation, which could influence nucleosomes and other protein-DNA interactions. PMID- 15548522 TI - The rates of switching movement of troponin T between three states of skeletal muscle thin filaments determined by fluorescence resonance energy transfer. AB - Troponin (Tn) plays the key roles in the regulation of striated muscle contraction. Tn consists of three subunits (TnT, TnC, and TnI). In combination with the stopped-flow method, fluorescence resonance energy transfer between probes attached to Cys-60 or Cys-250 of TnT and Cys-374 of actin was measured to determine the rates of switching movement of the troponin tail domain (Cys-60) and of the TnT-TnI coiled-coil C terminus (Cys-250) between three states (relaxed, closed, and open) of the thin filament. When the free Ca(2+) concentration was rapidly changed, these domains moved with rates of approximately 450 and approximately 85 s(-1) at pH 7.0 on Ca(2+) up and down, respectively. When myosin subfragment 1 (S1) was dissociated from thin filaments by rapid mixing with ATP, these domains moved with a single rate constant of approximately 400 s(-1) in the presence and absence of Ca(2+). The light scattering measurements showed that ATP-induced S1 dissociation occurred with a rate constant >800 s(-1). When S1 was rapidly mixed with the thin filament, these domains moved with almost the same or slightly faster rates than those of S1 binding measured by light scattering. In most but not all aspects, the rates of movement of the troponin tail domain and of the TnT-TnI coiled-coil C terminus were very similar to those of certain TnI sites (N terminus, Cys-133, and C terminus) previously characterized (Shitaka, Y., Kimura, C., Iio, T., and Miki, M. (2004) Biochemistry 43, 10739-10747), suggesting that a series of conformational changes in the Tn complex during switching on or off process occurs synchronously. PMID- 15548523 TI - Critical role of mitochondrial glutathione in the survival of hepatocytes during hypoxia. AB - Hypoxia is known to stimulate reactive oxygen species (ROS) generation. Because reduced glutathione (GSH) is compartmentalized in cytosol and mitochondria, we examined the specific role of mitochondrial GSH (mGSH) in the survival of hepatocytes during hypoxia (5% O2). 5% O2 stimulated ROS in HepG2 cells and cultured rat hepatocytes. Mitochondrial complex I and II inhibitors prevented this effect, whereas inhibition of nitric oxide synthesis with Nomega-nitro-L arginine methyl ester hydrochloride or the peroxynitrite scavenger uric acid did not. Depletion of GSH stores in both cytosol and mitochondria enhanced the susceptibility of HepG2 cells or primary rat hepatocytes to 5% O2 exposure. However, this sensitization was abrogated by preventing mitochondrial ROS generation by complex I and II inhibition. Moreover, selective mGSH depletion by (R,S)-3-hydroxy-4-pentenoate that spared cytosol GSH levels sensitized rat hepatocytes to hypoxia because of enhanced ROS generation. GSH restoration by GSH ethyl ester or by blocking mitochondrial electron flow at complex I and II rescued (R,S)-3-hydroxy-4-pentenoate-treated hepatocytes to hypoxia-induced cell death. Thus, mGSH controls the survival of hepatocytes during hypoxia through the regulation of mitochondrial generation of oxidative stress. PMID- 15548524 TI - Inhibition of muscarinic receptor-linked phospholipase D activation by association with tubulin. AB - Mammalian phospholipase D (PLD) is considered a key enzyme in the transmission signals from various receptors including muscarinic receptors. PLD activation is a rapid and transient process, but a negative regulator has not been found that inhibits signal-dependent PLD activation. Here, for the first time, we report that tubulin binding to PLD2 is an inhibition mechanism for muscarinic receptor linked PLD2 activation. Tubulin was identified in an immunoprecipitated PLD2 complex from COS-7 cells by peptide mass fingerprinting. The direct interaction between PLD2 and tubulin was found to be mediated by a specific region of PLD2 (amino acids 476-612). PLD2 was potently inhibited (IC50 <10 nM) by tubulin binding in vitro. In cells, the interaction between PLD2 and tubulin was increased by the microtubule disrupting agent nocodazole and reduced by the microtubule stabilizing agent Taxol. Moreover, PLD2 activity was found to be inversely correlated with the level of monomeric tubulin. In addition, we found that interaction with and the inhibition of PLD2 by monomeric tubulin is important for the muscarinic receptor-linked PLD signaling pathway. Interaction between PLD2 and tubulin was increased only after 1-2 min of carbachol stimulation when carbachol-stimulated PLD2 activity was decreased. The expression of the tubulin binding region of PLD2 blocked the later decrease in carbachol induced PLD activity by masking tubulin binding. Taken together, these results indicate that an increase in local membrane monomeric tubulin concentration inhibits PLD2 activity, and provides a novel mechanism for the inhibition of muscarinic receptor-induced PLD2 activation by interaction with tubulin. PMID- 15548525 TI - The unique C-terminal tail of the mitogen-activated protein kinase ERK5 regulates its activation and nuclear shuttling. AB - ERK5 is unique among mitogen-activated protein kinases (MAPKs) in that it contains a large C-terminal tail. We addressed the question of how this tail could affect the signaling capacity of ERK5. Gradual deletion of the C-terminal domains resulted in a drastic increase of ERK5 kinase activity, which was dependent on the up-stream MAPK cascade, thus indicating a possible auto inhibitory function of the tail. It is interesting that ERK5 was able to autophosphorylate its own tail. Moreover, ERK5, which was found to be expressed in virtually all kinds of cell lines, localized to nuclear as well as cytoplasmic compartments. The localization of ERK5 was determined by its C-terminal domains, which were also required for appropriate nucleocytoplasmic shuttling. Taken together, these results indicate that ERK5 signaling is directed by the presence of its unique C-terminal tail, which might be the key to understanding the key role of ERK5 in MAPK signaling. PMID- 15548526 TI - Site-directed mutagenesis of CC chemokine receptor 1 reveals the mechanism of action of UCB 35625, a small molecule chemokine receptor antagonist. AB - The chemokine receptor CCR1 and its principal ligand, CCL3/MIP-1alpha, have been implicated in the pathology of several inflammatory diseases including rheumatoid arthritis, multiple sclerosis, and asthma. As such, these molecules are the focus of much research with the ultimate aim of developing novel therapies. We have described previously a non-competitive small molecule antagonist of CCR1 (UCB 35625), which we hypothesized interacted with amino acids located within the receptor transmembrane (TM) helices (Sabroe, I., Peck, M. J., Jan Van Keulen, B., Jorritsma, A., Simmons, G., Clapham, P. R., Williams, T. J., and Pease, J. E. (2000) J. Biol. Chem. 275, 25985-25992). Here we describe an approach to identifying the mechanism by which the molecule antagonizes CCR1. Thirty-three point mutants of CCR1 were expressed transiently in L1.2 cells, and the cells were assessed for their capacity to migrate in response to CCL3 in the presence or absence of UCB 35625. Cells expressing the mutant constructs Y41A (TM helix 1, or TM1), Y113A (TM3), and E287A (TM7) were responsive to CCL3 but resistant to the antagonist, consistent with a role for the TM helices in CCR1 interactions with UCB 35625. Subsequent molecular modeling successfully docked the compound with CCR1 and suggests that the antagonist ligates TM1, 2, and 7 of CCR1 and severely impedes access to TM2 and TM3, a region thought to be perturbed by the chemokine amino terminus during the process of receptor activation. Insights into the mechanism of action of these compounds may facilitate the development of more potent antagonists that show promise as future therapeutic agents in the treatment of inflammatory disease. PMID- 15548527 TI - Aclacinomycin 10-hydroxylase is a novel substrate-assisted hydroxylase requiring S-adenosyl-L-methionine as cofactor. AB - Aclacinomycin 10-hydroxylase is a methyltransferase homologue that catalyzes a S adenosyl-L-methionine (AdoMet)-dependent hydroxylation of the C-10 carbon atom of 15-demethoxy-epsilon-rhodomycin, a step in the biosynthesis of the polyketide antibiotic beta-rhodomycin. S-Adenosyl-L-homocysteine is an inhibitor of the enzyme, whereas the AdoMet analogue sinefungin can act as cofactor, indicating that a positive charge is required for catalysis. 18O2 experiments show that the hydroxyl group is derived from molecular oxygen. The reaction further requires thiol reagents such as glutathione or dithiothreitol. Incubation of the enzyme with substrate in the absence of reductant leads to the accumulation of an intermediate with a molecular mass consistent with a perhydroxy compound. This intermediate is turned into product upon addition of glutathione. The crystal structure of an abortive enzyme-AdoMet product ternary complex reveals large conformational changes consisting of a domain rotation leading to active site closure upon binding of the anthracycline ligand. The data suggest a mechanism where decarboxylation of the substrate results in the formation of a carbanion intermediate, which is stabilized by resonance through the aromatic ring system of the anthracycline substrate. The delocalization of the electrons is facilitated by the positive charge of the cofactor AdoMet. The activation of oxygen and formation of a hydroxyperoxide intermediate occurs in a manner similar to that observed in flavoenzymes. Aclacinomycin-10-hydroxylase is the first example of a AdoMet-dependent hydroxylation reaction, a novel function for this cofactor. The enzyme lacks methyltransferase activity due to the positioning of the AdoMet methyl group unfavorable for a SN2-type methyl transfer to the substrate. PMID- 15548528 TI - Aluminum triggers decreased aconitase activity via Fe-S cluster disruption and the overexpression of isocitrate dehydrogenase and isocitrate lyase: a metabolic network mediating cellular survival. AB - Although aluminum is known to be toxic to most organisms, its precise biochemical interactions are not fully understood. In the present study, we demonstrate that aluminum promotes the inhibition of aconitase (Acn) activity via the perturbation of the Fe-S cluster in Pseudomonas fluorescens. Despite the significant decrease in citrate isomerization activity, cellular survival is assured by the overexpression of isocitrate lyase and isocitrate dehydrogenase (IDH)-NADP+. 13C NMR spectroscopic studies, Blue Native PAGE, and Western blot analyses indicated that although the decrease in Acn activity is concomitant with the increase of aluminum in the culture, the amount of Acn expressed is not sensitive to the concentration of the trivalent metal. A 6-fold decrease in Acn activity and no discernable change in protein content in aluminum-stressed cultures were observed. The addition of Fe(NH4)2(SO4)2 in a reducing environment led to a significant recovery in Acn activity. This enzymatic activity reverted to normal levels when aluminum-stressed cells were transferred to either a control or an iron-supplemented medium. The overexpression of the two isocitrate-metabolizing enzymes isocitrate lyase and IDH-NADP+ appears to mitigate the deficit in Acn activity. The levels of these enzymes are dependent on the aluminum content of the culture and appear to be under transcriptional control. Hence, the regulation of the enzymes involved in the homeostasis of isocitrate constitutes a pivotal component of the global metabolic strategy that ensures the survival of this organism in an aluminum citrate environment. PMID- 15548529 TI - Functional comparison of mouse slc26a6 anion exchanger with human SLC26A6 polypeptide variants: differences in anion selectivity, regulation, and electrogenicity. AB - The unusually low 78% amino acid identity between the orthologous human SLC26A6 and mouse slc26a6 polypeptides prompted systematic comparison of their anion transport functions in Xenopus oocytes. Multiple human SLC26A6 variant polypeptides were also functionally compared. Transport was studied as unidirectional fluxes of (36)Cl(-), [(14)C]oxalate, and [(35)S]sulfate; as net fluxes of HCO(3)(-) by fluorescence ratio measurement of intracellular pH; as current by two-electrode voltage clamp; and as net Cl(-) flux by fluorescence intensity measurement of relative changes in extracellular and intracellular [Cl( )]. Four human SLC26A6 polypeptide variants each exhibited rates of bidirectional [(14)C]oxalate flux, Cl(-)/HCO(3)(-) exchange, and Cl(-)/OH(-) exchange nearly equivalent to those of mouse slc26a6. Cl(-)/HCO(3)(-) exchange by both orthologs was cAMP-sensitive, further enhanced by coexpressed wild type cystic fibrosis transmembrane regulator but inhibited by cystic fibrosis transmembrane regulator DeltaF508. However, the very low rates of (36)Cl(-) and [(35)S]sulfate transport by all active human SLC26A6 isoforms contrasted with the high rates of the mouse ortholog. Human and mouse orthologs also differed in patterns of acute regulation. Studies of human-mouse chimeras revealed cosegregation of the high (36)Cl(-) transport phenotype with the transmembrane domain of mouse slc26a6. Mouse slc26a6 and human SLC26A6 each mediated electroneutral Cl(-)/HCO(3)(-) and Cl(-)/OH(-) exchange. In contrast, whereas Cl(-)/oxalate exchange by mouse slc26a6 was electrogenic, that mediated by human SLC26A6 appeared electroneutral. The increased currents observed in oocytes expressing either mouse or human ortholog were pharmacologically distinct from the accompanying monovalent anion exchange activities. The human SLC26A6 polypeptide variants SLC26A6c and SLC26A6d were inactive as transporters of oxalate, sulfate, and chloride. Thus, the orthologous mouse and human SLC26A6 proteins differ in anion selectivity, transport mechanism, and acute regulation, but both mediate electroneutral Cl( )/HCO(3)(-) exchange. PMID- 15548530 TI - Jingzhaotoxin-I, a novel spider neurotoxin preferentially inhibiting cardiac sodium channel inactivation. AB - Jingzhaotoxin-I (JZTX-I), a 33-residue polypeptide, is derived from the Chinese tarantula Chilobrachys jing-zhao venom based on its ability to evidently increase the strength and the rate of vertebrate heartbeats. The toxin has three disulfide bonds with the linkage of I-IV, II-V, and III-VI that is a typical pattern found in inhibitor cystine knot molecules. Its cDNA determined by rapid amplification of 3'- and 5'-cDNA ends encoded a 62-residue precursor with a small proregion of eight residues. Whole-cell configuration indicated that JZTX-I was a novel neurotoxin preferentially inhibiting cardiac sodium channel inactivation by binding to receptor site 3. Although JZTX-I also exhibits the interaction with channel isoforms expressing in mammalian and insect sensory neurons, its affinity for tetrodotoxin-resistant subtype in mammalian cardiac myocytes (IC50 = 31.6 nm) is approximately 30-fold higher than that for tetrodotoxin-sensitive subtypes in latter tissues. Not affecting outward delay-rectified potassium channels expressed in Xenopus laevis oocytes and tetrodotoxin-resistant sodium channels in mammal sensory neurons, JZTX-I hopefully represents a potent ligand to discriminate cardiac sodium channels from neuronal tetrodotoxin-resistant isoforms. Furthermore, different from any reported spider toxins, the toxin neither modifies the current-voltage relationships nor shifts the steady-state inactivation of sodium channels. Therefore, JZTX-I defines a new subclass of spider sodium channel toxins. JZTX-I is an alpha-like toxin first reported from spider venoms. The result provides an important witness for a convergent functional evolution between spider and other animal venoms. PMID- 15548531 TI - A pantothenate kinase from Staphylococcus aureus refractory to feedback regulation by coenzyme A. AB - The key regulatory step in CoA biosynthesis in bacteria and mammals is pantothenate kinase (CoaA), which governs the intracellular concentration of CoA through feedback regulation by CoA and its thioesters. CoaA from Staphylococcus aureus (SaCoaA) has a distinct primary sequence that is more similar to the mammalian pantothenate kinases than the prototypical bacterial CoaA of Escherichia coli. In contrast to all known pantothenate kinases, SaCoaA activity is not feedback-regulated by CoA or CoA thioesters. Metabolic labeling of S. aureus confirms that CoA levels are not controlled by CoaA or at steps downstream from CoaA. The pantothenic acid antimetabolite N-heptylpantothenamide (N7-Pan) possesses potent antimicrobial activity against S. aureus and has multiple cellular targets. N7-Pan is a substrate for SaCoaA and is converted to the inactive butyldethia-CoA analog by the downstream pathway enzymes. The analog is also incorporated into acyl carrier protein and D-alanyl carrier protein, the prosthetic groups of which are derived from CoA. The inactivation of acyl carrier protein and the cessation of fatty acid synthesis are the most critical causes of growth inhibition by N7-Pan because the toxicity of the drug is ameliorated by supplementing the growth medium with fatty acids. The absence of feedback regulation at the pantothenate kinase step allows the accumulation of high concentrations of intracellular CoA, consistent with the physiology of S. aureus, which lacks glutathione and relies on the CoA/CoA disulfide reductase redox system for protection from oxidative damage. PMID- 15548532 TI - Genetic evidence for a multi-subunit complex in coenzyme Q biosynthesis in yeast and the role of the Coq1 hexaprenyl diphosphate synthase. AB - Coenzyme Q (Q) is a lipid that functions as an electron carrier in the mitochondrial respiratory chain in eukaryotes. There are eight complementation groups of Q-deficient Saccharomyces cerevisiae mutants designated coq1-coq8. Here we provide genetic evidence that several of the Coq polypeptides interact with one another. Deletions in any of the COQ genes affect the steady-state expression of Coq3p, Coq4p, and Coq6p. Antibodies that recognize Coq1p, a hexaprenyl diphosphate synthase, were generated and used to determine that Coq1p is peripherally associated with the inner membrane on the matrix side. Yeast Deltacoq1 mutants harboring diverse Coq1 orthologs from prokaryotic species produce distinct sizes of polyprenyl diphosphate and hence distinct isoforms of Q including Q(7), Q(8), Q(9), or Q(10) (Okada, K., Kainou, T., Matsuda, H., and Kawamukai, M. (1998) FEBS Lett. 431, 241-244). We find that steady-state levels of Coq3p, Coq4p, and Coq6p are rescued in some cases to near wild-type levels by the presence of these diverse Coq1 orthologs in the Deltacoq1 mutant. These data suggest that the lipid product of Coq1p or a Q-intermediate derived from polyprenyl diphosphate is involved in stabilizing the Coq3, Coq4, and Coq6 polypeptides. PMID- 15548533 TI - Genomic organization and functional analysis of murine PKD2L1. AB - Mutations in genes that encode polycystins 1 or 2 cause polycystic kidney disease (PKD). Here, we report the genomic organization and functional expression of murine orthologue of human polycystin-2L1 (PKD2L1). The murine PKD2L1 gene comprises 15 exons in chromosome 19C3. Coexpression of PKD2L1 together with polycystin-1 (PKD1) resulted in the expression of PKD2L1 channels on the cell surface, whereas PKD2L1 expressed alone was retained within the endoplasmic reticulum (ER). This suggested that interaction between PKD1 and PKD2L1 is essential for PKD2L1 trafficking and channel formation. Deletion analysis at the cytoplasmic tail of PKD2L1 revealed that the coiled-coil domain was important for trafficking by PKD1. Mutagenesis within two newly identified ER retention signal like amino acid sequences caused PKD2L1 to be expressed at the cell surface. This indicated that the coiled-coil domain was responsible for retaining PKD2L1 within the ER. Functional analysis of murine PKD2L1 expressed in HEK 293 cells was undertaken using calcium imaging. Coexpression of PKD1 and PKD2L1 resulted in the formation of functional cation channels that were opened by hypo-osmotic stimulation, whereas neither molecule formed functional channels when expressed alone. We conclude that PKD2L1 forms functional cation channels on the plasma membrane by interacting with PKD1. These findings raise the possibility that PKD2L1 represents the third genetic locus that is responsible for PKD. PMID- 15548534 TI - Identification of NdhL and Ssl1690 (NdhO) in NDH-1L and NDH-1M complexes of Synechocystis sp. PCC 6803. AB - The subunit compositions of two types of NAD(P)H dehydrogenase complexes of Synechocystis sp. PCC 6803, NDH-1L and NDH-1M, were studied by two-dimensional blue-native/SDS-PAGE followed by electrospray tandem mass spectrometry. Fifteen proteins were observed in NDH-1L including hydrophilic subunits (NdhH, -K, -I, J, -M, and -N) and hydrophobic subunits (NdhA, -B, -E, -G, -D1, and -F1). In addition, NdhL and a novel subunit, Ssl1690 (designated NdhO), were shown to be components of this complex. All subunits mentioned above were present in the NDH 1M complex except NdhD1 and NdhF1. NdhL and Ssl1690 (NdhO) were homologous to hypothetical proteins encoded by genomic DNA in higher plants, suggesting that chloroplast NDH-1 complexes contain related subunits. Diagnostic sequence motifs were found for both NdhL and NdhO homologous proteins. Analysis of ndhL deletion mutant (M9) revealed the presence of assembled NDH-1L and NDH-1M complexes, but these complexes appear to be functionally impaired in the absence of NdhL. Both NDH-1 complexes were absent in the ndhB deletion mutant (M55). PMID- 15548535 TI - Mapping the rho1 GABA(C) receptor agonist binding pocket. Constructing a complete model. AB - Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian brain. The GABA receptor type C (GABA(C)) is a ligand-gated ion channel with pharmacological properties distinct from the GABA(A) receptor. To date, only three binding domains in the recombinant rho1 GABA(C) receptor have been recognized among six potential regions. In this report, using the substituted cysteine accessibility method, we scanned three potential regions previously unexplored in the rho1 GABA(C) receptor, corresponding to the binding loops A, E, and F in the structural model for ligand-gated ion channels. The cysteine accessibility scanning and agonist/antagonist protection tests have resulted in the identification of residues in loops A and E, but not F, involved in forming the GABA(C) receptor agonist binding pocket. Three of these newly identified residues are in a novel region corresponding to the extended stretch of loop E. In addition, the cysteine accessibility pattern suggests that part of loop A and part of loop E have a beta-strand structure, whereas loop F is a random coil. Finally, when all of the identified ligand binding residues are mapped onto a three-dimensional homology model of the amino-terminal domain of the rho1 GABA(C) receptor, they are facing toward the putative binding pocket. Combined with previous findings, a complete model of the GABA(C) receptor binding pocket was proposed and discussed in comparison with the GABA(A) receptor binding pocket. PMID- 15548536 TI - In vitro phosphorylation by cAMP-dependent protein kinase up-regulates recombinant Saccharomyces cerevisiae mannosylphosphodolichol synthase. AB - DPM1 is the structural gene for mannosylphosphodolichol synthase (i.e. Dol-P-Man synthase, DPMS) in Saccharomyces cerevisiae. Earlier studies with cDNA cloning and sequence analysis have established that 31-kDa DPMS of S. cerevisiae contains a consensus sequence (YRRVIS141) that can be phosphorylated by cAMP-dependent protein kinase (PKA). We have been studying the up-regulation of DPMS activity by protein kinase A-mediated phosphorylation in higher eukaryotes, and used the recombinant DPMS from S. cerevisiae in this study to advance our knowledge further. DPMS catalytic activity was indeed enhanced severalfold when the recombinant protein was phosphorylated in vitro. The rate as well as the magnitude of catalysis was higher with the phosphorylated enzyme. A similar increase in the catalytic activity was also observed when the in vitro phosphorylated recombinant DPMS was assayed as a function of increasing concentrations of exogenous dolichylmonophosphate (Dol-P). Kinetic studies indicated that there was no change in the Km for GDP-mannose between the in vitro phosphorylated and control recombinant DPMS, but the Vmax was increased by 6-fold with the phosphorylated enzyme. In vitro phosphorylated recombinant DPMS also exhibited higher enzyme turnover (kcat) and enzyme efficiency (kcat/Km). SDS-PAGE followed by autoradiography of the 32P-labeled DPMS detected a 31-kDa phosphoprotein, and immunoblotting with anti-phosphoserine antibody established the presence of a phosphoserine residue in in vitro phosphorylated recombinant DPMS. To confirm the phosphorylation activation of recombinant DPMS, serine 141 in the consensus sequence was replaced with alanine by PCR site-directed mutagenesis. The S141A DPMS mutant exhibited more than half-a-fold reduction in catalytic activity compared with the wild type when both were analyzed after in vitro phosphorylation. Thus, confirming that S. cerevisiae DPMS activity is indeed regulated by the cAMP-dependent protein phosphorylation signal, and the phosphorylation target is serine 141. PMID- 15548537 TI - Type I PDZ ligands are sufficient to promote rapid recycling of G Protein-coupled receptors independent of binding to N-ethylmaleimide-sensitive factor. AB - Molecular sorting of G protein-coupled receptors (GPCRs) between divergent recycling and lysosomal pathways determines the functional consequences of agonist-induced endocytosis. The carboxyl-terminal cytoplasmic domain of the beta2 adrenergic receptor (beta2AR) mediates both PDZ binding to Na+/H+ exchanger regulatory factor/ezrin/radixin/moesin-binding phosphoprotein of 50 kDa (NHERF/EBP50) family proteins and non-PDZ binding to the N-ethylmaleimide sensitive factor (NSF). We have investigated whether PDZ interaction(s) are actually sufficient to promote rapid recycling of endocytosed receptors and, if so, whether PDZ-mediated sorting is restricted to the beta2AR tail or to sequences that bind NHERF/EBP50. The trafficking effects of short (10 residue) sequences differing in PDZ and NSF binding properties were examined using chimeric mutant receptors. The recycling activity of the beta2AR-derived tail sequence was not blocked by a point mutation that selectively disrupts binding to NSF, and naturally occurring PDZ ligand sequences were identified that do not bind detectably to NSF yet function as strong recycling signals. The carboxyl terminal cytoplasmic domain of the beta1-adrenergic receptor, which does not bind either to NSF or NHERF/EBP50 and interacts selectively with a distinct group of PDZ proteins, promoted rapid recycling of chimeric mutant receptors with efficiency similarly high as that of the beta2AR tail. These results indicate that PDZ domain-mediated protein interactions are sufficient to promote rapid recycling of GPCRs, independent of binding to NSF. They also suggest that PDZ directed recycling is a rather general mechanism of GPCR regulation, which is not restricted to a single GPCR, and may involve additional PDZ domain-containing protein(s) besides NHERF/EBP50. PMID- 15548538 TI - Fragile X-related protein FXR1P regulates proinflammatory cytokine tumor necrosis factor expression at the post-transcriptional level. AB - Tumor necrosis factor (TNF) is regulated post-transcriptionally by the AU-rich element (ARE) within the 3'-untranslated region of its mRNA. This regulation modulates translational efficacy and mRNA stability. By using a cRNA probe containing the TNF ARE sequence, we screened a macrophage protein expression library and identified FXR1P. Macrophages that we generated from FXR1 knock-out mice had enhanced TNF protein production compared with wild type macrophages following activation. Expression of several other proteins that are regulated by ARE sequences was also affected by FXR1P deficiency. A GFP-ARE reporter that has green fluorescent protein (GFP) expression under control of the 3'-untranslated region of TNF mRNA had enhanced expression in transfected macrophages deficient in FXR1P. Finally, we found that the ablation of FXR1P led to a dramatically enhanced association of the TNF mRNA with polyribosomes demonstrating the important role of FXR1P in the post-transcriptional regulation of TNF expression. Our data suggest that release of this repression by FXR1P occurs during lipopolysaccharide-induced macrophage activation. Finally, complementation of the knock-out macrophages with recombinant FXR1P resulted in decreased TNF protein production, supporting our findings that FXR1P operates as a repressor of TNF translation. PMID- 15548539 TI - A dynamic zinc redox switch. AB - The crystal structures of glutathione-dependent formaldehyde-activating enzyme (Gfa) from Paracoccus denitrificans, which catalyzes the formation of S hydroxymethylglutathione from formaldehyde and glutathione, and its complex with glutathione (Gfa-GTT) have been determined. Gfa has a new fold with two zinc sulfur centers, one that is structural (zinc tetracoordinated) and one catalytic (zinc apparently tricoordinated). In Gfa-GTT, the catalytic zinc is displaced due to disulfide bond formation of glutathione with one of the zinc-coordinating cysteines. Soaking crystals of Gfa-GTT with formaldehyde restores the holoenzyme. Accordingly, the displaced zinc forms a complex by scavenging formaldehyde and glutathione. The activation of formaldehyde and of glutathione in this zinc complex favors the final nucleophilic addition, followed by relocation of zinc in the catalytic site. Therefore, the structures of Gfa and Gfa-GTT draw the critical association between a dynamic zinc redox switch and a nucleophilic addition as a new facet of the redox activity of zinc-sulfur sites. PMID- 15548540 TI - Spermine causes loss of innate immune response to Helicobacter pylori by inhibition of inducible nitric-oxide synthase translation. AB - Helicobacter pylori infection of the stomach elicits a vigorous but ineffective host immune and inflammatory response, resulting in persistence of the bacterium for the life of the host. We have reported that in macrophages, H. pylori up regulates inducible NO synthase (iNOS) and antimicrobial NO production, but in parallel there is induction of arginase II, generating ornithine, and of ornithine decarboxylase (ODC), generating polyamines. Spermine, in particular, has been shown to restrain immune response in activated macrophages by inhibiting proinflammatory gene expression. We hypothesized that spermine could prevent the antimicrobial effects of NO by inhibiting iNOS in macrophages activated by H. pylori. Spermine did not affect the up-regulation of iNOS mRNA levels but in a concentration-dependent manner significantly attenuated iNOS protein levels and NO production. Reduction in iNOS protein was due to inhibition of iNOS translation and not due to iNOS degradation. ODC knockdown with small interfering (si) RNA resulted in increased H. pylori-stimulated iNOS protein expression and NO production without altering iNOS mRNA levels. When macrophages were cocultured with H. pylori, killing of bacteria was enhanced by transfection of ODC siRNA and prevented by addition of spermine. These results identify a mechanism of immune dysregulation induced by H. pylori in which stimulated spermine synthesis by the arginase-ODC pathway inhibits iNOS translation and NO production, leading to persistence of the bacterium and risk for peptic ulcer disease and gastric cancer. PMID- 15548541 TI - Crystal structure of thrombin bound to heparin. AB - Thrombin is the final protease in the blood coagulation cascade and serves both pro- and anticoagulant functions through the cleavage of several targets. The ability of thrombin to specifically recognize a wide range of substrates derives from interactions that occur outside of the active site of thrombin. Thrombin possesses two anion binding exosites, which mediate many of its interactions with cofactors and substrates, and although many structures of thrombin have been solved, few such interactions have been described in molecular detail. Glycosaminoglycan binding to exosite II of thrombin plays a major role in switching off the procoagulant functions of thrombin by mediating its irreversible inhibition by circulating serpins and by its binding to the endothelial cell surface receptor thrombomodulin. Here we report the 1.85-A structure of human alpha-thrombin bound to a heparin fragment of eight monosaccharide units in length. The asymmetric unit is composed of two thrombin dimers, each sharing a single heparin octasaccharide chain. The observed interactions are fully consistent with previous mutagenesis studies and illustrate on a molecular level the cofactor interaction that is critical for the restriction of clotting to the site of blood vessel injury. PMID- 15548542 TI - Sphingolipid uptake by cultured cells: complex aggregates of cell sphingolipids with serum proteins and lipoproteins are rapidly catabolized. AB - Human fibroblasts, rat neurons, and murine neuroblastoma cells, cultured in the presence of fetal calf serum, were fed with [1-(3)H]sphingosine to radiolabel sphingolipids. The fate of cell sphingolipids, the release of sphingolipids in the culture medium, the interaction of sphingolipids with the proteins and lipoproteins of fetal calf serum, and the fate of sphingolipids taken up by the cells were investigated. For this latter purpose, the culture medium containing radioactive sphingolipids was delivered to nonlabeled cells. The presence of tritium at position 1 of sphingosine allowed us to follow the extent of sphingolipid catabolism by measuring the production of radioactive phosphatidylethanolamine and proteins by recycling the radioactive ethanolamine formed during sphingosine catabolism and the production of tritiated water. We confirmed that in cells the recycling of sphingosine occurred to a high extent and that only a minor portion of cell sphingolipids was catabolized to the small fragments of ethanolamine and water. Cell sphingolipids were released in the culture medium, where they formed large lipoproteic aggregates at a rate of about 12% per day. Released sphingolipids were taken up by the cells and catabolized to the sphingosine and then to ethanolamine, and recycling of sphingosine was not observed. This suggests that in the presence of fetal calf serum in the culture medium, exogenous sphingolipids directly reach the lysosomes, were they are entirely catabolized. Thus, the trafficking of sphingolipids from cells to the extracellular environment and from this to other cells does not allow the modification of the plasma membrane composition. PMID- 15548543 TI - The optimal measure of linkage disequilibrium reduces error in association mapping of affection status. AB - We have developed a simple yet powerful approach for disease gene association mapping by linkage disequilibrium (LD). This method is unique because it applies a model with evolutionary theory that incorporates a parameter for the location of the causal polymorphism. The method exploits LD maps, which assign a location in LD units (LDU) for each marker. This approach is based on single marker tests within a composite likelihood framework, which avoids the heavy Bonferroni correction through multiple testing. As a proof of principle, we tested an 890 kb region flanking the CYP2D6 gene associated with poor drug-metabolizing activity in order to refine the localization of a causal mutation. Previous LD mapping studies using single markers and haplotypes have identified a 390 kb significant region associated with the poor drug-metabolizing phenotype on chromosome 22. None of the 27 Single nucleotide polymorphisms was within the gene. Using a metric LDU map, the commonest functional polymorphism within the gene was located at 14.9 kb from its true location, surrounded within a 95% confidence interval of 172 kb. The kb map had a relative efficiency of 33% compared with the LDU map. Our findings indicate that the support interval and location error are smaller than any published results. Despite the low resolution and the strong LD in the region, our results provide evidence of the substantial utility of LDU maps for disease gene association mapping. These tests are robust to large numbers of markers and are applicable to haplotypes, diplotypes, whole-genome association or candidate region studies. PMID- 15548544 TI - Genome-wide linkage scan of epilepsy-related photoparoxysmal electroencephalographic response: evidence for linkage on chromosomes 7q32 and 16p13. AB - Photoparoxysmal response (PPR) is an abnormal visual sensitivity of the brain in reaction to intermittent photic stimulation. It is an epilepsy-related electroencephalographic trait with high prevalence in idiopathic epilepsies, especially in common idiopathic generalized epilepsies (IGEs), such as childhood absence epilepsy and juvenile myoclonic epilepsy. This degree of co-morbidity suggests that PPR may be involved in the predisposition to IGE. The identification of genes for PPR would, therefore, aid the dissection of the genetic basis of IGE. Sixteen PPR-multiplex families were collected to conduct a genome-wide linkage scan using broad (all PPR types) and narrow (exclusion of PPR types I and II and the occipital epilepsy cases) models of affectedness for PPR. We found an empirical genome-wide significance for parametric (HLOD) and non parametric (NPL) linkage (Pgw(HLOD)=0.004 and Pgw(NPL)=0.01) for two respective chromosomal regions, 7q32 at D7S1804 (HLOD=3.47 with alpha=1, P(NPL)=3.39x10(-5)) and 16p13 at D16S3395 (HLOD=2.44 with alpha=1, P(NPL)=7.91x10(-5)). These two genomic regions contain genes that are important for the neuromodulation of cortical dynamics and may represent good targets for candidate-gene studies. Our study identified two susceptibility loci for PPR, which may be related to the underlying myoclonic epilepsy phenotype present in the families studied. PMID- 15548545 TI - Mouse model carrying H222P-Lmna mutation develops muscular dystrophy and dilated cardiomyopathy similar to human striated muscle laminopathies. AB - Laminopathies are a group of disorders caused by mutations in the LMNA gene encoding A-type lamins, components of the nuclear lamina. Three of these disorders affect specifically the skeletal and/or cardiac muscles, and their pathogenic mechanisms are still unknown. We chose the LMNA H222P missense mutation identified in a family with autosomal dominant Emery-Dreifuss muscular dystrophy, one of the striated muscle-specific laminopathies, to create a faithful mouse model of this type of laminopathy. The mutant mice exhibit overtly normal embryonic development and sexual maturity. At adulthood, male homozygous mice display reduced locomotion activity with abnormal stiff walking posture and all of them die by 9 months of age. As for cardiac phenotype, they develop chamber dilation and hypokinesia with conduction defects. These abnormal skeletal and cardiac features were also observed in the female homozygous mice but with a later-onset than in males. Histopathological analysis of the mice revealed muscle degeneration with fibrosis associated with dislocation of heterochromatin and activation of Smad signalling in heart and skeletal muscles. These results demonstrate that LmnaH222P/H222P mice represent a good model for studying laminopathies affecting striated muscles as they develop a dystrophic condition of both skeletal and cardiac muscles similar to the human diseases. PMID- 15548546 TI - Abnormalities of social interactions and home-cage behavior in a mouse model of Rett syndrome. AB - Rett syndrome (RTT) is an autistic spectrum disorder with a known genetic basis. RTT is caused by loss of function mutations in the X-linked gene MECP2 and is characterized by loss of acquired motor, social and language skills in females beginning at 6-18 months of age. MECP2 mutations also cause non-syndromic mental retardation in males and females, and abnormalities of MeCP2 expression in the brain have been found in autistic spectrum disorders. We studied home-cage behavior and social interactions in a mouse model of RTT (Mecp2(308/Y)) carrying a mutation similar to common RTT causing alleles. Young adult mutant mice showed abnormal home-cage diurnal activity in the absence of motor skill deficits. Nesting, a phenotype related to social behavior, and social interactions were both impaired in these animals. Mecp2(308/Y) mice showed deficits in nest building and decreased nest use. Although there were no differences in aggression or exploration of novel inanimate stimuli, mutant mice took less initiative and were less decisive approaching unfamiliar males and spent less time in close vicinity to them in several social interaction paradigms. The abnormalities of diurnal activity and social behavior in Mecp2(308/Y) mice are reminiscent of the sleep/wake dysfunction and autistic features of RTT. These data suggest that MECP2 regulates the expression and/or function of genes involved in social behavior. The study of Mecp2(308/Y) mice will allow the identification of the molecular basis of social impairment in RTT and related autistic spectrum disorders. PMID- 15548547 TI - TTF-1 and RET promoter SNPs: regulation of RET transcription in Hirschsprung's disease. AB - Single nucleotide polymorphisms (SNPs) of the coding regions of receptor tyrosine kinase gene (RET) are associated with Hirschsprung's disease (HSCR, aganglionic megacolon). These SNPs, individually or combined, may act as a low penetrance susceptibility locus and/or be in linkage disequilibrium (LD) with another susceptibility locus located in RET regulatory regions. Because two RET promoter SNPs have been found associated with HSCR, in LD with HSCR-associated RET coding region haplotypes, their implication in the transcriptional regulation of RET is of major interest. Analysis of 172 sporadic HSCR patients also revealed the presence of HSCR-associated RET promoter SNPs in LD with the main coding region RET haplotype observed in Chinese patients. By using a weighted logistic regression approach, we determined that of all SNPs tested in our study, the promoter SNPs are the most correlated to the disease. Functional analysis of the RET promoter SNPs in the context of additional 5' regulatory regions demonstrated that the HSCR-associated alleles decrease RET transcription. These SNPs overlap a TTF-1 binding site and TTF-1-activated RET transcription is also decreased by the HSCR-associated SNPs. Moreover, we identified an HSCR patient with a Gly322Ser TTF-1 mutation that compromises activation of transcription from HSCR-associated RET promoter haplotypes. Interestingly, we show that the pattern of RET and TTF-1 expression is coincident in developing human gut. We also present a detailed profile of the RET gene in our population, which provides an insight into the higher incidence of the disease in China. PMID- 15548548 TI - Transcriptional dysregulation in striatal projection- and interneurons in a mouse model of Huntington's disease: neuronal selectivity and potential neuroprotective role of HAP1. AB - Transcriptional dysregulation has been described as a central mechanism in the pathogenesis of Huntington's disease (HD), in which medium spiny projection neurons (MSN) selectively degenerate whereas neuronal nitric-oxide-synthase positive interneurons (nNOS-IN) survive. In order to begin to understand this differential vulnerability we compared mRNA levels of selected genes involved in N-methyl-D-aspartate (NMDA) glutamate receptor and calcium (Ca2+) signaling pathways in MSN and nNOS-IN from 12-week-old R6/2 mice, a transgenic mouse model of HD and wild-type littermates. We undertook a laser capture microdissection (LCM) study to examine the contribution of transcriptional dysregulation in candidate genes involved in these two signaling pathways in discrete populations of striatal neurons. The use of LCM in combination with quantitative real-time polymerase chain reaction (Q-PCR) allowed us to quantify the neuronal abundance of candidate mRNAs. We found different transcriptional alterations in R6/2 neurons for both MSN and nNOS-IN, indicating that global transcriptional dysregulation alone does not account for selective vulnerability. Further, we observed a striking enrichment of several mRNAs in the nNOS-IN population, including that for the NMDA receptor subunit NR2D, the postsynaptic density protein 95 (PSD-95) and the huntingtin-associated protein 1 (HAP1) as well as nitric-oxide-synthase (nNOS) mRNA itself. The higher expression levels of these molecules in nNOS-IN when compared with MSN together with an association of nNOS, NR2D and HAP1 in a protein complex with PSD-95 suggest that these proteins may be involved in protective pathways that contribute to the resistance of this interneuron population to neurodegeneration in HD. PMID- 15548549 TI - Transgenic mouse model of early-onset DYT1 dystonia. AB - Early-onset dystonia is an autosomal dominant movement disorder associated with deletion of a glutamic acid residue in torsinA. We generated four independent lines of transgenic mice by overexpressing human DeltaE-torsinA using a neuron specific enolase promoter. The transgenic mice developed abnormal involuntary movements with dystonic-appearing, self-clasping of limbs, as early as 3 weeks after birth. Animals also showed hyperkinesia and rapid bi-directional circling. Approximately 40% of transgenic mice from each line demonstrated these severe behavioral abnormalities. Neurochemical analyses revealed decreases in striatal dopamine in affected transgenic mice, although levels were increased in those that had no behavioral changes. Immunohistochemistry demonstrated perinuclear inclusions and aggregates that stained positively for ubiquitin, torsinA and lamin, a marker of the nuclear envelope. Inclusions were detected in neurons of the pedunculopontine nucleus and in other brain stem regions in a pattern similar to what has been described in DYT1 patients. This transgenic mouse model demonstrates behavioral and pathologic features similar to patients with early onset dystonia and may help to better understand the pathophysiology of this disorder and to develop more effective therapies. PMID- 15548550 TI - Surviving CA1 pyramidal cells receive intact perisomatic inhibitory input in the human epileptic hippocampus. AB - Temporal lobe epilepsy (TLE) is known to be linked to an impaired balance of excitation and inhibition. Whether inhibition is decreased or preserved in the human epileptic hippocampus, beside the excess excitation, is still a debated question. In the present study, quantitative light and electron microscopy has been performed to analyse the distribution, morphology and input-output connections of parvalbumin (PV)-immunopositive interneurons, together with the entire perisomatic input of pyramidal cells, in the human control and epileptic CA1 region. Based on the degree of cell loss, the patients with therapy-resistant TLE formed four pathological groups. In the non-sclerotic CA1 region of TLE patients, where large numbers of pyramidal cells are preserved, the number of PV immunopositive cell bodies decreased, whereas axon terminal staining, and the distribution of their postsynaptic targets was not altered. The synaptic coverage of CA1 pyramidal cell axon initial segments (AISs) remained unchanged in the epileptic tissue. The somatic inhibitory input is also preserved; it has been decreased only in the cases with patchy pyramidal cell loss in the CA1 region (control, 0.637; epileptic with mild cell loss, 0.642; epileptic with patchy cell loss, 0.424 microm synaptic length/100 microm soma perimeter). The strongly sclerotic epileptic CA1 region, where pyramidal cells can hardly be seen, contains a very small number of PV-immunopositive elements. Our results suggest that perisomatic inhibitory input is preserved in the epileptic CA1 region as long as pyramidal cells are present. Basket and axo-axonic cells survive in epilepsy if their original targets are present, although many of them lose their PV content or PV immunoreactivity. An efficient perisomatic inhibition is likely to take part in the generation of abnormal synchrony in the non-sclerotic epileptic CA1 region, and thus participate in the maintenance of epileptic seizures driven, for example, by hyperactive afferent input. PMID- 15548551 TI - A neural basis for collecting behaviour in humans. AB - Collecting behaviour is commonplace in the normal population, but there has been little investigation of its neural basis in humans. The observation that collecting behaviour can assume pathological proportions in patients with certain patterns of brain damage led us to hypothesize that dysfunction in a system encompassing mesial prefrontal cortices accounts for abnormal collecting and may guide normal collecting. We tested the hypothesis in 86 subjects with focal lesions of the telencephalon, by relating the neuroanatomical placement of the lesions to the presence of repetitive and indiscriminate acquisition behaviour and impaired discard behaviour. The subjects had no history of psychiatric disease or abnormal collecting behaviour prior to lesion onset. Lesions were analysed with high-resolution three-dimensional MRI. Collecting behaviour was evaluated with a standardized questionnaire completed by a close relative of each subject. Thirteen subjects exhibited abnormal collecting, characterized by massive and disruptive accumulation of useless objects. In all cases, the abnormality of collecting behaviour was severe and persisted despite attempted interventions and obvious negative consequences. There were no differences between pathological collectors and non-collectors on tests of executive functions or anterograde memory. All subjects with pathological collecting behaviour had damage to the mesial frontal region (including the right polar sector and the anterior cingulate), but there was no damage to most of the subcortical structures that, in species such as rodents, are known to drive the acquisition and retention of objects. The evidence suggests that damage to the mesial frontal region disrupts a mechanism which normally modulates subcortically driven predispositions to acquire and collect, and adjusts these predispositions to environmental context. PMID- 15548552 TI - Combination of 'idiopathic' REM sleep behaviour disorder and olfactory dysfunction as possible indicator for alpha-synucleinopathy demonstrated by dopamine transporter FP-CIT-SPECT. AB - REM sleep behaviour disorder (RBD) and olfactory dysfunction are common and very early features of alpha-synucleinopathies, in particular Parkinson's disease. To investigate the hypothesis that these two clinical features in combination are an indicator of evolving alpha-synucleinopathy, olfactory function was assessed in RBD. We studied 30 patients (18 male, 12 female; mean age 48 +/- 14 years, range 19-78 years) with clinical (idiopathic, n = 6; symptomatic, n = 13, mostly associated with narcolepsy) or subclinical (n = 11, associated with narcolepsy) RBD according to standard criteria and 30 age- and gender-matched healthy control subjects using standardized 'Sniffin' Sticks'. RBD patients had a significantly higher olfactory threshold (P = 0.0001), lower discrimination score (P = 0.003), and lower identification score (P = 0.001). Compared with normative data, 97% of the RBD patients had a pathologically increased olfactory threshold, 63% an impaired odour discrimination score, and 63% a decreased identification score. On neurological examination, signs of parkinsonism were newly found in five patients with clinical RBD (not associated with narcolepsy), who usually had a long history of 'idiopathic' RBD. Four of the five patients fulfilled the UK Brain Bank criteria for the clinical diagnosis of Parkinson's disease. The underlying nigrostriatal degeneration of clinical Parkinson's disease was confirmed by I-123 FP-CIT SPECT in one patient and early nigrostriatal degeneration was identified by SPECT in a further two patients with 'idiopathic' clinical RBD out of 11 RBD patients who agreed to undergo SPECT studies. Our study shows that RBD patients have a profound impairment of olfactory function. Five patients with clinical RBD not associated with narcolepsy had clinical or imaging signs of nigrostriatal degeneration. This new clinical finding correlates with the neuropathological staging of Parkinson's disease (stages 1-3) as proposed by Braak. In stage 1, the anterior olfactory nucleus or the olfactory bulb is affected (along with the dorsal motor nucleus of the glossopharyngeal and vagal nerves). In stage 2, additional lesions consistently remain confined to the medulla oblongata and pontine tegmentum, which are critical areas for RBD. Midbrain lesions are found only in stage 3, in particular degeneration of dopaminergic neurons in the substantia nigra pars compacta. Thus, 'idiopathic' RBD patients with olfactory impairment might present with stage 2 preclinical alpha-synucleinopathy. Since narcoleptic patients are not known to have an increased risk of developing parkinsonism, the pathophysiology and clinical relevance of hyposmia in RBD/narcolepsy patients requires further research. PMID- 15548553 TI - Cognitive sequelae of head injury: involvement of basal forebrain and associated structures. AB - Traumatic brain injury is the most common cause of death and disability in young people and survivors often suffer from chronic cognitive deficits. From animal, post-mortem and cognitive studies, there is now increased evidence that abnormalities in the cholinergic system may be underlying some of these deficits. This study investigated this hypothesis in a group of survivors of moderate severe head injury (n = 31). Patients completed a comprehensive neuropsychological assessment and an MRI scan. Compared with a group of controls (matched on age, sex and premorbid intelligence quotient), the patients showed deficits in sustained attention, paired associate learning and reaction time, but comparative preservation of spatial working memory. Voxel-based morphometry revealed reduced grey matter density in the head injured group in the basal forebrain, the hippocampal formation and regions of the neocortex. These cognitive and structural results are consistent with cholinergic dysfunction. These preliminary findings suggest that cholinergic enhancers may be an effective treatment of cognitive deficits post head injury. PMID- 15548554 TI - Abstract and concrete concepts have structurally different representational frameworks. AB - The architecture supporting our conceptual knowledge of abstract words has remained almost entirely unexplored. By contrast, a vast neuropsychological, neurolinguistic and neuroimaging literature has addressed questions relating to the structure of the semantic system underpinning our knowledge of concrete items (e.g. artefacts and animals). In the context of semantic refractory access dysphasia, a series of experiments exploring and comparing abstract and concrete word comprehension are described. We demonstrate that semantically associated abstract words reliably interfere with one another significantly more than semantically synonymous abstract words, while concrete words show the reverse pattern. We report the first evidence that abstract and concrete word meanings are based in representational systems that have qualitatively different properties. More specifically, we show that abstract concepts, but not concrete concepts, are represented in an associative neural network. Furthermore, our patient was found to have significantly greater difficulty in identifying high frequency than low frequency abstract words. This observation constitutes the first evidence of an inverse word frequency effect. Our results challenge the generality of many existing models of human conceptual knowledge, which derive their structure from experimental findings in the concrete domain alone. PMID- 15548555 TI - Illusory persistence of touch after right parietal damage: neural correlates of tactile awareness. AB - We studied a patient who experienced 'palinaesthesia', an illusion of persistent touch following tactile stimulation on the left hand, subsequent to a right parietal meningioma affecting primary somatosensory regions in the postcentral gyrus (SI) and superior parietal gyrus (Brodmann area 7), but preserving the secondary somatosensory cortex (SII) in the upper lateral sulcus. This subjective sensation was accompanied by transient increases in objective measures of tactile threshold. The patient had mild deficits in superficial tactile perception, but showed severe left-sided extinction for offsets of tactile stimuli during bilateral stimulation, but not for onsets of stimuli. Functional MRI revealed increased neural activity during palinaesthesia selectively arising within the ipsilesional-right SI cortex, but no abnormality within left SI and bilateral SII. Right SI responded to the onset of new tactile stimuli on the left hand but not to their offset. By contrast, any tactile events on either hand modulated activity in contralateral SII regions, even undetected left-sided offsets. These data demonstrate that illusory persistence of touch following stimulation on the hand may result from sustained neural activity in a restricted region of the SI cortex outlasting the offset of the actual tactile stimuli. These findings also provide direct evidence for a critical role of SI in mediating conscious somatosensory experience on contralateral parts of the body. PMID- 15548556 TI - A novel autosomal recessive myopathy with external ophthalmoplegia linked to chromosome 17p13.1-p12. AB - We describe a new autosomal recessive myopathy of early onset and very slow progression distinguished by the prominent external ophthalmoplegia in 16 subjects of eight families from a large and highly inbred Arab community. Characteristic clinical features include mild facial and skeletal muscle weakness and atrophy more pronounced proximally in the upper limbs, facial dysmorphism and scoliosis associated with conjugate, non-restrictive ocular motility impairment greatest in the upgaze and without ptosis or aberrant eye movements. Orbital MRI in the patients demonstrated atrophy with fatty replacement of the oculorotatory muscles. The major pathological alteration on skeletal muscle biopsy was a marked type 1 fibre predominance with core-like formations. A genome wide search for regions of homozygosity in the affected members from two informative families identified linkage with chromosome 17p13.1-p12 markers. Maximum two-point logarithm of odds scores were obtained at loci D17S1803 and AFMA070WD1 (Zmax = 3.74 at = 0). Two independent recombination events at D17S1812 and D17S947 further defined a critical region of 12 cM. Several genes map to this interval, including a cluster of sarcomeric myosin heavy chain genes. One of these genes, MYH2, is involved in inclusion body myopathy 3, but no exonic mutations were found by direct sequencing. The molecular basis for this new myopathy remains to be identified. PMID- 15548557 TI - Mapping the brain in autism. A voxel-based MRI study of volumetric differences and intercorrelations in autism. AB - Autism is a disorder of neurodevelopment resulting in pervasive abnormalities in social interaction and communication, repetitive behaviours and restricted interests. There is evidence for functional abnormalities and metabolic dysconnectivity in 'social brain' circuitry in this condition, but its structural basis has proved difficult to establish reliably. Explanations for this include replication difficulties inherent in 'region of interest' approaches usually adopted, and variable inclusion criteria for subjects across the autism spectrum. Moreover, despite a consensus that autism probably affects widely distributed brain regions, the issue of anatomical connectivity has received little attention. Therefore, we planned a fully automated voxel-based whole brain volumetric analysis in children with autism and normal IQ. We predicted that brain structural changes would be similar to those previously shown in adults with autism spectrum disorder and that a correlation analysis would suggest structural dysconnectivity. We included 17 stringently diagnosed children with autism and 17 age-matched controls. All children had IQ >80. Using Brain Activation and Morphological Mapping (BAMM) software, we measured global brain and tissue class volumes and mapped regional grey and white matter differences across the whole brain. With the expectation that volumes of interconnected regions correlate positively, we carried out a preliminary exploration of 'connectivity' in autism by comparing the nature of inter-regional grey matter volume correlations with control. Children with autism had a significant reduction in total grey matter volume and significant increase in CSF volume. They had significant localized grey matter reductions within fronto-striatal and parietal networks similar to findings in our previous study, and additional decreases in ventral and superior temporal grey matter. White matter was reduced in the cerebellum, left internal capsule and fornices. Correlation analysis revealed significantly more numerous and more positive grey matter volumetric correlations in controls compared with children with autism. Thus, using similar diagnostic criteria and image analysis methods in otherwise healthy populations with an autistic spectrum disorder from different countries, cultures and age groups, we report a number of consistent findings. Taken together, our data suggest abnormalities in the anatomy and connectivity of limbic-striatal 'social' brain systems which may contribute to the brain metabolic differences and behavioural phenotype in autism. PMID- 15548558 TI - Microanatomy of the dysplastic neocortex from epileptic patients. AB - Focal cortical dysplasia (FCD) is a pathology that is characterized by the abnormal development of the neocortex. Indeed, a wide range of abnormalities in the cortical mantle have been associated with this pathology, including cytoarchitectonic alterations and the presence of dysmorphic neurons, balloon cells and ectopic neurons in the white matter. FCD is commonly associated with epilepsy, and hence we have studied the ultrastructure of cortical tissue resected from three subjects with intractable epilepsy secondary to cortical dysplasia to identify possible alterations in synaptic circuitry, using correlative light and electron microscopic methods. While the balloon cells found in this tissue do not appear to receive synaptic contacts, the ectopic neurons in the white matter were abnormally large and were surrounded by hypertrophic basket formations immunoreactive for the calcium-binding protein parvalbumin. Furthermore, these basket formations formed symmetrical (inhibitory) synapses with both the somata and the proximal portion of the dendrites of these giant ectopic neurons. A quantitative analysis revealed that in the dysplastic tissue, the density of excitatory and inhibitory synapses was different from that of the normal adjacent cortex. Both increases and decreases in synaptic density were observed, as well as changes in the proportion of excitatory and inhibitory synapses. However, we could not establish a common pattern of changes, either in the same patients or between different patients. These results suggest that cortical dysplasia leads to multiple changes in excitatory and inhibitory synaptic circuits. We discuss the possible relationship between these alterations and epilepsy, bearing in mind the possible limitations that preclude the extrapolation of the results to the whole population of epileptic patients with dysplastic neocortex. PMID- 15548559 TI - Pathological study of spinal cord atrophy in multiple sclerosis suggests limited role of local lesions. AB - Imaging studies in multiple sclerosis have shown that spinal cord atrophy correlates with clinical disability. The pathological substrate of atrophy has not as yet been investigated adequately. In order to determine the cause of spinal cord atrophy in multiple sclerosis, five different sections of the spinal cord were examined histopathologically in 33 controls and 55 multiple sclerosis cases. In the multiple sclerosis cases in each section the total lesion load and the cross-sectional area of the cord were measured. Multiple regression models were estimated, controlling for sex, age, duration of the disease and location of the cord sections. The multiple sclerosis cords were found to be significantly smaller than the controls. The duration of the disease played the most important role in determining cord atrophy. The degree of atrophy varied in different parts of the cord. Individual lesions played a minor role in local atrophy. Our findings suggest that axonal degeneration, possibly caused by the cumulative number of lesions in the brain and cord, or an alternative atrophic process, is responsible for spinal cord atrophy in multiple sclerosis, rather than tissue loss within individual lesions. PMID- 15548560 TI - Socioeconomic inequalities in indicator scores for diabetes: poor quality or poor measures? PMID- 15548561 TI - Association of deprivation, ethnicity, and sex with quality indicators for diabetes: population based survey of 53,000 patients in primary care. AB - OBJECTIVES: To determine the effect of deprivation and ethnicity on the achievement of quality indicators for patients with diabetes and the extent of any inequalities between the sexes. DESIGN: Population based cross sectional survey using electronic general practice records. SETTING: 237 UK practices contributing to the QRESEARCH database. PARTICIPANTS: 54,180 patients with diabetes, derived from a population of 1.8 million patients. MAIN OUTCOME MEASURES: Adjusted odds ratios for 18 indicators for diabetes from the new general medical services contract for UK general practitioners and comparisons between patients from the most deprived and most affluent fifths (areas of high and low ethnicity) and between men and women. RESULTS: The prevalence of diabetes was 3.0%, and there was a large variation between practices in achievement of indicators. Compared with patients from affluent areas, those from deprived areas were less likely to have body mass index and smoking status recorded. They were also less likely to have records for HbA1c concentration; an HbA1c value < 7.5% or < 10%; retinal screening; blood pressure; testing for neuropathy or microalbuminuria, or flu vaccination. Compared with patients from areas of low ethnicity those from areas of high ethnicity were less likely to have many measures recorded. Women were significantly less likely to have records for body mass index; pulses; blood pressure values below 145/85 mm Hg; testing for microalbuminuria; serum cholesterol concentration; serum cholesterol values < 5 mmol/l; and angiotensin converting enzyme inhibitors given in the presence of proteinuria or microalbuminuria. CONCLUSIONS: Practices in areas of high deprivation and high ethnicity will have to work harder to achieve the quality indicators for diabetes, and it is possible that those practices that most need the resources are the ones least likely to get them. PMID- 15548562 TI - New signs of a strengthening U.S. nurse labor market? AB - Wage increases, relatively high national unemployment, and widespread private sector initiatives aimed at increasing the number of people who become nurses has resulted in a second straight year of strong employment growth among registered nurses (RNs). In 2003, older women and, to a lesser extent, foreign-born RNs accounted for a large share of employment growth. We also observe unusually large employment growth from two new demographic groups: younger people, particularly women in their early thirties, and men. Yet, despite the increase in employment of nearly 185,000 hospital RNs since 2001, the evidence suggests that the current nurse shortage has not been eliminated. PMID- 15548563 TI - Microalbuminuria in adults after prenatal exposure to the Dutch famine. AB - Maternal undernutrition during gestation is associated with an increase in cardiovascular risk factors in the offspring in adult life. The effect of famine exposure during different stages of gestation on adult microalbuminuria (MA) was studied. MA was measured in 724 people, aged 48 to 53, who were born as term singletons in a university hospital in Amsterdam, the Netherlands, around the time of the Dutch famine 1944 to 1945. Twelve percent of people who were exposed to famine in mid gestation had MA (defined as albumin/creatinine ratio >/=2.5) compared with 7% of those who were not prenatally exposed to famine (odds ratio 2.1; 95% confidence interval 1.0 to 4.3). Correcting for BP, diabetes, and other influences that affect MA did not attenuate this association (adjusted odds ratio 3.2; 95% confidence interval 1.4 to 7.7). The effect of famine was independent of size at birth. Midgestation is a period of rapid increase in nephron number, which is critical in determining nephron endowment at birth. Fetal undernutrition may lead to lower nephron endowment with consequent MA in adult life. PMID- 15548564 TI - Glomerular podocytopathy in patients with systemic lupus erythematosus. AB - A series of patients with systemic lupus erythematosus (SLE) and proteinuria were studied to determine whether nephrotic-range proteinuria was associated with diffuse epithelial cell foot process effacement in the absence of peripheral glomerular immune aggregate deposition. Biopsies from patients with known or suspected SLE and a histologic diagnosis of (1) normal by light microscopy, (2) mesangial proliferative glomerulonephritis, or (3) focal segmental glomerulosclerosis were studied. Biopsies were excluded when they demonstrated endocapillary proliferation or necrosis by light microscopy or electron-dense glomerular basement membrane deposits by electron microscopy. Patients were required to fulfill four of 11 American Rheumatologic Association criteria for the diagnosis of SLE, and proteinuria could not be associated with nonsteroidal anti-inflammatory drug use. Eighteen biopsies were studied, eight from patients with nephrotic-range proteinuria (>/=3 g/d) and 10 from patients with non nephrotic proteinuria. The time from diagnosis of SLE to biopsy was shorter for nephrotic patients that for nonnephrotic patients. Seven of eight biopsies from nephrotic patients demonstrated at least 80% foot process effacement, whereas no biopsy from a nonnephrotic patient exhibited >20% effacement. There were no other significant pathologic differences between the nephrotic and nonnephrotic patients. The single common morphologic feature associated with nephrotic proteinuria was diffuse visceral epithelial cell foot process effacement. It is concluded that the development of nephrotic-range proteinuria in patients with SLE without peripheral immune aggregate deposition or endocapillary proliferation on renal biopsy is more likely a manifestation of SLE than the coexistence of idiopathic minimal-change glomerulopathy and SLE. PMID- 15548565 TI - Characterization of the regulation and functional consequences of p21ras activation in neutrophils by antineutrophil cytoplasm antibodies. AB - Antineutrophil cytoplasm antibodies (ANCA) are implicated in the pathogenesis of systemic vasculitis. ANCA are directed against antigens expressed on the surface of cytokine-primed neutrophils. It was shown previously that whole IgG ANCA and its fraction antigen binding [F(ab')(2)] fragment can activate the GTPase p21(ras). This study shows a functional involvement of this molecule in the ANCA activation of neutrophils by inhibiting the production of superoxide with farnesylthiosalicylic acid. Using the ras activation assay, farnesylthiosalicylic acid inhibits p21(ras) binding to its substrate at comparable concentrations to those seen for superoxide inhibition. It is also shown that activation of p21(ras) by ANCA is transient, peaking at 5 to 10 min and returning to baseline by 30 min. The use of ras isoform-specific antibodies in Western blots established, for the first time, that Harvey-ras is not present in human neutrophils, but both Kirsten-ras (K-ras) and Neuronal-ras are. Stimulation with ANCA is able to differentially activate K-ras without effects on neuronal-ras. The activation of p21(ras) by ANCA and its F(ab')(2) is prevented by inhibition of both Src kinases and phosphatidylinositol-3-kinase, indicating a cooperative role for both molecules in the G protein pathway activated by ANCA F(ab')(2) upstream of p21(ras). It is concluded that ANCA selectively activates K-ras during induction of a respiratory burst via pathways involving multiple upstream kinases. PMID- 15548566 TI - Alteration of product specificity of Aeropyrum pernix farnesylgeranyl diphosphate synthase (Fgs) by directed evolution. AB - Directed evolution of the C25 farnesylgeranyl diphosphate synthase of Aeropyrum pernix (Fgs) was carried out by error-prone PCR with an in vivo color complementation screen utilizing carotenoid biosynthetic pathway enzymes. Screening yielded 12 evolved clones with C20 geranylgeranyl diphosphate synthase activity which were isolated and characterized in order to understand better the chain elongation mechanism of this enzyme. Analysis of these mutants revealed three different mechanisms of product chain length specificity. Two mutants (A64T and A64V) have a single mutation at the 8th amino acid upstream of a conserved first aspartate-rich motif (FARM), which is involved in the mechanism for chain elongation reaction of all prenyl diphosphate synthases. One mutant (A135T) carries a single mutation at the 7th amino acid upstream of another conserved region (141GQ142), which was recently found to be another important region controlling chain elongation of a type III C20 geranylgeranyl diphosphate synthase and Escherichia coli C15 farnesyl diphosphate synthase. Finally, one mutant carrying four mutations (V84I, H88R, I177 M and M191V) is of interest. Molecular modeling, site-directed mutagenesis and in vitro assays of this mutant suggest that product chain-length distribution can be also controlled by a structural change provoked by a cooperative interaction of amino acids. PMID- 15548567 TI - Improved growth response of antibody/receptor chimera attained by the engineering of transmembrane domain. AB - Structure-based design of antibody/cytokine receptor chimeras has permitted a growth signal transduction in response to non-natural ligands such as fluorescein conjugated BSA as mimicry of cytokine-cytokine receptor systems. However, while tight on/off regulation is observed in the natural cytokine receptor systems, many chimeras constructed to date showed residual growth-promoting activity in the absence of ligands. Here we tried to reduce the basal growth signal intensity from a chimera by engineering the transmembrane domain (TM) that is thought to be involved in the interchain interaction of natural cytokine receptors. When the retroviral vectors encoding the chimeras with either the wild-type erythropoietin receptor (EpoR) TM or the one bearing two mutations in the leucine zipper motif were transduced to non-strictly interleukin-6-dependent 7TD1 cells, a tight antigen-dependent on/off regulation was attained, also demonstrating the first antigen-mediated genetically modified cell amplification of non-strictly factor dependent cells. The results clearly indicate that the TM mutation is an effective means to improve the growth response of the antibody/receptor chimera. PMID- 15548568 TI - Molecular determinants of voltage-gated sodium channel regulation by the Nedd4/Nedd4-like proteins. AB - The voltage-gated Na(+) channels (Na(v)) form a family composed of 10 genes. The COOH termini of Na(v) contain a cluster of amino acids that are nearly identical among 7 of the 10 members. This COOH-terminal sequence, PPSYDSV, is a PY motif known to bind to WW domains of E3 protein-ubiquitin ligases of the Nedd4 family. We recently reported that cardiac Na(v)1.5 is regulated by Nedd4-2. In this study, we further investigated the molecular determinants of regulation of Na(v) proteins. When expressed in HEK-293 cells and studied using whole cell voltage clamping, the neuronal Na(v)1.2 and Na(v)1.3 were also downregulated by Nedd4-2. Pull-down experiments using fusion proteins bearing the PY motif of Na(v)1.2, Na(v)1.3, and Na(v)1.5 indicated that mouse brain Nedd4-2 binds to the Na(v) PY motif. Using intrinsic tryptophan fluorescence imaging of WW domains, we found that Na(v)1.5 PY motif binds preferentially to the fourth WW domain of Nedd4-2 with a K(d) of approximately 55 muM. We tested the binding properties and the ability to ubiquitinate and downregulate Na(v)1.5 of three Nedd4-like E3s: Nedd4 1, Nedd4-2, and WWP2. Despite the fact that along with Nedd4-2, Nedd4-1 and WWP2 bind to Na(v)1.5 PY motif, only Nedd4-2 robustly ubiquitinated and downregulated Na(v)1.5. Interestingly, coexpression of WWP2 competed with the effect of Nedd4 2. Finally, using brefeldin A, we found that Nedd4-2 accelerated internalization of Na(v)1.5 stably expressed in HEK-293 cells. This study shows that Nedd4 dependent ubiquitination of Na(v) channels may represent a general mechanism regulating the excitability of neurons and myocytes via modulation of channel density at the plasma membrane. PMID- 15548569 TI - Expression levels of RyR1 and RyR3 control resting free Ca2+ in skeletal muscle. AB - To better understand the role of the transient expression of ryanodine receptor (RyR) type 3 (RyR3) on Ca(2+) homeostasis during the development of skeletal muscle, we have analyzed the effect of expression levels of RyR3 and RyR1 on the overall physiology of cultured myotubes and muscle fibers. Dyspedic myotubes were infected with RyR1 or RyR3 containing virions at 0.2, 0.4, 1.0, and 4.0 moieties of infection (MOI), and analysis of their pattern of expression, caffeine sensitivity, and resting free Ca(2+) concentration ([Ca(2+)](r)) was performed. Although increased MOI resulted in increased expression of each receptor isoform, it did not significantly affect the immunopattern of RyRs or the expression levels of calsequestrin, triadin, or FKBP-12. Interestingly, myotubes expressing RyR3 always had significantly higher [Ca(2+)](r) and lower caffeine EC(50) than did cells expressing RyR1. Although some of the increased sensitivity of RyR3 to caffeine could be attributed to the higher [Ca(2+)](r) in RyR3-expressing cells, studies of [(3)H]ryanodine binding demonstrated intrinsic differences in caffeine sensitivity between RyR1 and RyR3. Tibialis anterior (TA) muscle fibers at different stages of postnatal development exhibited a transient increase in [Ca(2+)](r) coordinately with their level of RyR3 expression. Similarly, adult soleus fibers, which also express RyR3, had higher [Ca(2+)](r) than did adult TA fibers, which exclusively express RyR1. These data show that in skeletal muscle, RyR3 increases [Ca(2+)](r) more than RyR1 does at any expression level. These data suggest that the coexpression of RyR1 and RyR3 at different levels may constitute a novel mechanism by which to regulate [Ca(2+)](r) in skeletal muscle. PMID- 15548570 TI - Role of NBC1 in apical and basolateral HCO3- permeabilities and transendothelial HCO3- fluxes in bovine corneal endothelium. AB - Corneal transparency and hydration control are dependent on HCO(3)(-) transport properties of the corneal endothelium. Recent work (13) suggested the presence of an apical 1Na(+)-3HCO(3)(-) cotransporter (NBC1) in addition to a basolateral 1Na(+)-2HCO(3)(-) cotransporter. We examined whether the NBC1 cotransporter contributes significantly to basolateral or apical HCO(3)(-) permeability and whether the cotransporter participates in transendothelial net HCO(3)(-) flux in cultured bovine corneal endothelium. NBC1 protein expression was reduced using small interfering RNA (siRNA). Immunoblot analysis showed that 5-15 nM siRNA decreased NBC1 expression by 80-95%, 4 days posttransfection. Apical and basolateral HCO(3)(-) permeabilities were determined by measuring the rate of pH(i) change when HCO(3)(-) was removed from the bath under constant pH or constant CO(2) conditions. Using either protocol, we found that cultures treated with NBC1 siRNA had sixfold lower basolateral HCO(3)(-) permeability than untreated or siCONTROL siRNA-treated cells. Apical HCO(3)(-) permeability was unaffected by NBC1 siRNA treatment. Net non-steady-state HCO(3)(-) flux was 0.707 +/- 0.009 mM.min(-1).cm(2) in the basolateral-to-apical direction and increased to 1.74 +/- 0.15 when cells were stimulated with 2 muM forskolin. Treatment with 5 nM siRNA decreased basolateral-to-apical flux by 67%, whereas apical-to basolateral flux was unaffected, significantly decreasing net HCO(3)(-) flux to 0.236 +/- 0.002. NBC1 siRNA treatment or 100 muM ouabain also eliminated steady state HCO(3)(-) flux, as measured by apical compartment alkalinization. Collectively, reduced basolateral HCO(3)(-) permeability, basolateral-to-apical fluxes, and net HCO(3)(-) flux as a result of reduced expression of NBC1 indicate that NBC1 plays a key role in transendothelial HCO(3)(-) flux and is functional only at the basolateral membrane. PMID- 15548571 TI - Mechanical loading and injury induce human myotubes to release neutrophil chemoattractants. AB - The purpose of this study was to 1) test the hypothesis that skeletal muscle cells (myotubes) after mechanical loading and/or injury are a source of soluble factors that promote neutrophil chemotaxis and superoxide anion (O(2)(-).) production and 2) determine whether mechanical loading and/or injury causes myotubes to release cytokines that are known to influence neutrophil responses [tumor necrosis factor-alpha (TNF-alpha), IL-8, and transforming growth factor beta1 (TGF-beta1)]. Human myotubes were grown in culture and exposed to either a cyclic strain (0, 5, 10, 20, or 30% strain) or a scrape injury protocol. Protocols of 5, 10, and 20% strain did not cause injury, whereas 30% strain and scrape injury caused a modest and a high degree of injury, respectively. Conditioned media from strained myotubes promoted chemotaxis of human blood neutrophils and primed them for O(2)(-). production in a manner that was dependent on a threshold of strain and independent from injury. Neutrophil chemotaxis, but not priming, progressively increased with higher magnitudes of strain. Conditioned media only from scrape-injured myotubes increased O(2)(-). production from neutrophils. Concentrations of IL-8 and total TGF-beta1 in conditioned media were reduced by mechanical loading, whereas TNF-alpha and active TGF-beta1 concentrations were unaffected. In conclusion, skeletal muscle cells after mechanical loading and injury are an important source of soluble factors that differentially influence neutrophil chemotaxis and the stages of neutrophil-derived reactive oxygen species production. Neutrophil responses elicited by mechanical loading, however, did not parallel changes in the release of IL-8, TGF-beta1, or TNF-alpha from skeletal muscle cells. PMID- 15548572 TI - Muscle-specific interaction of caveolin isoforms: differential complex formation between caveolins in fibroblastic vs. muscle cells. AB - It is generally well accepted that caveolin-3 expression is muscle specific, whereas caveolin-1 and -2 are coexpressed in a variety of cell types, including adipocytes, endothelial cells, epithelial cells, and fibroblasts. Caveolin-1 and 2 are known to form functional hetero-oligomeric complexes in cells where they are coexpressed, whereas caveolin-3 forms homo-oligomeric high molecular mass complexes. Although caveolin-2 might be expected to interact in a similar manner with caveolin-3, most studies indicate that this is not the case. However, this view has recently been challenged as it has been demonstrated that caveolin-2 and -3 are coexpressed in primary cultures of cardiac myocytes, where these two proteins can be coimmunoprecipitated. Thus it remains controversial whether caveolin-2 interacts with caveolin-3. Here, we directly address the issue of caveolin isoform protein-protein interactions by means of three distinct molecular genetic approaches. First, using caveolin-1-deficient mouse embryonic fibroblasts, in which we have stably expressed caveolin-1, -2, or -3, we find that caveolin-1 interacts with caveolin-2 in this setting, whereas caveolin-3 does not, in agreement with most published observations. Next, we used a transfected L6 myoblast cell system expressing all three caveolin proteins. Surprisingly, we found that caveolin-1, -2, and -3 all coimmunoprecipitate in this cell type, suggesting that this interaction is muscle cell specific. Similar results were obtained when the skeletal muscle of caveolin-1 transgenic animals was analyzed for caveolin-1 and caveolin-3 coimmunoprecipitation. Thus we conclude that all three caveolins can interact to form a discrete hetero oligomeric complex, but that such complex formation is clearly muscle specific. PMID- 15548573 TI - Targeting leukocyte integrins in human diseases. AB - As our understanding of integrins as multifunctional adhesion and signaling molecules has grown, so has their recognition as potential therapeutic targets in human diseases. Leukocyte integrins are of particular interest in this regard, as they are key molecules in immune-mediated and inflammatory processes and are thus critically involved in diverse clinical disorders, ranging from asthma to atherosclerosis. Antagonists that interfere with integrin-dependent leukocyte trafficking and/or post-trafficking events have shown efficacy in multiple preclinical models, but these have not always predicted success in subsequent clinical trials (e.g., ischemia-reperfusion disorders and transplantation). However, recent successes of integrin antagonists in psoriasis, inflammatory bowel disease, and multiple sclerosis demonstrate the tremendous potential of antiadhesion therapy directed at leukocyte integrins. This article will review the role of the leukocyte integrins in the inflammatory process, approaches to targeting leukocyte integrins and their ligands, and the results of completed clinical trials. PMID- 15548574 TI - Reduction in CD1d expression on dendritic cells and macrophages by an acute virus infection. AB - Mice were infected with lymphocytic choriomeningitis virus (LCMV) to determine if changes in CD1d expression occurred during an acute virus infection. It is interesting that a decrease in CD1d expression on splenic dendritic cells (DC) and macrophages (MPhi) was observed for at least 3 months post-LCMV infection, and vaccinia virus and vesicular stomatitis virus induced similar changes in CD1d upon infection with those viruses. The reduction of CD1d cell-surface expression on DC and MPhi was independent of interferon-gamma and interleukin-12 expression but partially recovered in transporter associated with antigen processing-1 deficient mice, suggesting that CD8+ T cells may play a role. Thus, one consequence of the induction of a cellular immune response is a change in CD1d expression, which may constitute a key element in regulating antiviral immunity. PMID- 15548575 TI - Age-related decline in RACK-1 expression in human leukocytes is correlated to plasma levels of dehydroepiandrosterone. AB - Aging is associated with remodeling of the immune system, contributing to increased incidence of infections, autoimmune diseases, and cancer among the elderly. Alterations in several signal transduction pathways have been reported to play an important role in immunosenescence. We show that peripheral blood leukocytes obtained from old donors (> or =65 years) have a significantly reduced expression of receptor for activated C kinase 1 (RACK-1), a protein required for protein kinase C (PKC)-beta signaling, as compared with young donors (< or =40 years), both in males and females. The decline in RACK-1 immunoboth in reactivity was age-related (Spearman correlation, r=-0.278, P=0.012). All leukocyte subpopulations, namely lympho-monocytes, granulocytes, and B and T cells, showed a similar defect. We also observed a direct correlation between circulating dehydroepiandrosterone (DHEA) and RACK-1 expression in leukocytes (Spearman correlation, r=0.388, P=0.001). Furthermore, in vitro treatment with DHEA resulted in increased RACK-1 expression in leukocytes and lymphocyte proliferation, confirming the role of this hormone in the modulation of its expression and immune functions. A relevant consequence of RACK-1-reduced expression was the observation that release of tumor necrosis factor alpha following lipopolysaccharide challenge and mitogen-induced lymphocye proliferation, which involves PKC-beta activation, was significantly reduced in elderly subjects. Overall, our findings contribute to the understanding of the complex process of immunosenescence and identify age-related loss in immunological responses as partially associated with decreased RACK-1 expression. PMID- 15548576 TI - Neural crest cell lineage segregation in the mouse neural tube. AB - Neural crest (NC) cells arise in the dorsal neural tube (NT) and migrate into the embryo to develop into many different cell types. A major unresolved question is when and how the fate of NC cells is decided. There is widespread evidence for multipotential NC cells, whose fates are decided during or after migration. There is also some evidence that the NC is already divided into subpopulations of discrete precursors within the NT. We have investigated this question in the mouse embryo. We find that a subpopulation of cells on the most dorsomedial aspect of the NT express the receptor tyrosine kinase Kit (previously known as c kit), emigrate exclusively into the developing dermis, and then express definitive markers of the melanocyte lineage. These are thus melanocyte progenitor cells. They are generated predominantly at the midbrain-hindbrain junction and cervical trunk, with significant numbers also in lower trunk. Other cells within the dorsal NT are Kit-, migrate ventrally, and, from embryonic day 9.5, express the neurotrophin receptor p75. These cells most likely only give rise to ventral NC derivatives such as neurons and glia. The p75+ cells are located ventrolateral to the Kit+ cells in areas of the NT where these two cell types are found. These data provide direct in vivo evidence for NC lineage segregation within the mouse neural tube. PMID- 15548577 TI - The COUP-TF nuclear receptors regulate cell migration in the mammalian basal forebrain. AB - Cells migrate via diverse pathways and in different modes to reach their final destinations during development. Tangential migration has been shown to contribute significantly to the generation of neuronal diversity in the mammalian telencephalon. GABAergic interneurons are the best-characterized neurons that migrate tangentially, from the ventral telencephalon, dorsally into the cortex. However, the molecular mechanisms and nature of these migratory pathways are only just beginning to be unravelled. In this study we have first identified a novel dorsal-to-ventral migratory route, in which cells migrate from the interganglionic sulcus, located in the basal telencephalon between the lateral and medial ganglionic eminences, towards the pre-optic area and anterior hypothalamus in the diencephalon. Next, with the help of transplantations and gain-of-function studies in organotypic cultures, we have shown that COUP-TFI and COUP-TFII are expressed in distinct and non-overlapping migratory routes. Ectopic expression of COUP-TFs induces an increased rate of cell migration and cell dispersal, suggesting roles in cellular adhesion and migration processes. Moreover, cells follow a distinct migratory path, dorsal versus ventral, which is dependent on the expression of COUP-TFI or COUP-TFII, suggesting an intrinsic role of COUP-TFs in guiding migrating neurons towards their target regions. Therefore, we propose that COUP-TFs are directly involved in tangential cell migration in the developing brain, through the regulation of short- and long range guidance cues. PMID- 15548578 TI - Defective paracrine signalling by TGFbeta in yolk sac vasculature of endoglin mutant mice: a paradigm for hereditary haemorrhagic telangiectasia. AB - Hereditary haemorrhagic telangiectasia (HHT) is an autosomal dominant disorder in humans that is characterised by multisystemic vascular dyplasia and recurrent haemorrhage. Germline mutations in one of two different genes, endoglin or ALK1 can cause HHT. Both are members of the transforming growth factor (TGF) beta receptor family of proteins, and are expressed primarily on the surface of endothelial cells (ECs). Mice that lack endoglin or activin receptor like kinase (ALK) 1 die at mid-gestation as a result of defects in the yolk sac vasculature. Here, we have analyzed TGFbeta signalling in yolk sacs from endoglin knockout mice and from mice with endothelial-specific deletion of the TGFbeta type II receptor (TbetaRII) or ALK5. We show that TGFbeta/ALK5 signalling from endothelial cells to adjacent mesothelial cells is defective in these mice, as evidenced by reduced phosphorylation of Smad2. This results in the failure of vascular smooth muscle cells to differentiate and associate with endothelial cells so that blood vessels remain fragile and become dilated. Phosphorylation of Smad2 and differentiation of smooth muscle can be rescued by culture of the yolk sac with exogenous TGFbeta1. Our data show that disruption of TGFbeta signalling in vascular endothelial cells results in reduced availability of TGFbeta1 protein to promote recruitment and differentiation of smooth muscle cells, and provide a possible explanation for weak vessel walls associated with HHT. PMID- 15548579 TI - EDEN-BP-dependent post-transcriptional regulation of gene expression in Xenopus somitic segmentation. AB - EDEN-BP is a Xenopus RNA-binding protein that triggers deadenylation [poly(A) tail shortening], and thereby translational repression and degradation, of a subset of maternal mRNAs soon after fertilization. We show here that this factor is expressed in the presomitic mesoderm of older embryos, the site where somitic segmentation takes place. Inhibiting EDEN-BP function using either antisense morpholino oligonucleotides or neutralizing antibodies leads to severe defects in somitic segmentation, but not myotomal differentiation. This is associated with defects in the expression of segmentation markers belonging to the Notch signalling pathway in the presomitic mesoderm. We show by a combination of approaches that the mRNA encoding XSu(H), a protein that plays a central role in Notch signalling, is regulated by the EDEN-BP pathway. Accordingly, XSu(H) is overexpressed in EDEN-BP knock-down embryos, and overexpressing XSu(H) causes segmentation defects. We finally give data indicating that, in addition to XSu(H), other segmentation RNAs are a target for EDEN-BP. These results show that EDEN-BP-dependent post-transcriptional regulation of gene expression is required for the process of somitic segmentation. PMID- 15548580 TI - Molecular dissection of Pax6 function: the specific roles of the paired domain and homeodomain in brain development. AB - The transcription factor Pax6 plays a key role during development of various organs, including the brain where it affects cell fate, cell proliferation and patterning. To understand how Pax6 coordinates these diverse effects at the molecular level, we examined the role of distinct DNA-binding domains of Pax6, the homeodomain (HD), the paired domain (PD) and its splice variant (5a), using loss- and gain-of-function approaches. Here we show that the PD is necessary for the regulation of neurogenesis, cell proliferation and patterning effects of Pax6, since these aspects are severely affected in the developing forebrain of the Pax6Aey18 mice with a deletion in the PD but intact homeo- and transactivation domains. In contrast, a mutation of the HD lacking DNA-binding (Pax64Neu) resulted in only subtle defects of forebrain development. We further demonstrate distinct roles of the two splice variants of the PD. Retrovirally mediated overexpression of Pax6 containing exon 5a inhibited cell proliferation without affecting cell fate, while Pax6 containing the canonical form of the PD lacking exon 5a affected simultaneously cell fate and proliferation. These results therefore demonstrate a key role of the PD in brain development and implicate splicing as a pivotal factor regulating the potent neurogenic role of Pax6. PMID- 15548581 TI - Planar cell polarity in the Drosophila eye is directed by graded Four-jointed and Dachsous expression. AB - Planar cell polarity (PCP) occurs when the cells of an epithelium are polarized along a common axis lying in the epithelial plane. During the development of PCP, cells respond to long-range directional signals that specify the axis of polarization. In previous work on the Drosophila eye, we proposed that a crucial step in this process is the establishment of graded expression of the cadherin Dachsous (Ds) and the Golgi-associated protein Four-jointed (Fj). These gradients were proposed to specify the direction of polarization by producing an activity gradient of the cadherin Fat within each ommatidium. In this report, I test and confirm the key predictions of this model by altering the patterns of Fj, Ds and Fat expression. It is shown that the gradients of Fj and Ds expression provide partially redundant positional information essential for specifying the polarization axis. I further demonstrate that reversing the Fj and Ds gradients can lead to reversal of the axis of polarization. Finally, it is shown that an ectopic gradient of Fat expression can re-orient PCP in the eye. In contrast to the eye, the endogenous gradients of Fj and Ds expression do not play a major role in directing PCP in the wing. Thus, this study reveals that the two tissues use different strategies to orient their PCP. PMID- 15548582 TI - Kaiso is a genome-wide repressor of transcription that is essential for amphibian development. AB - DNA methylation in animals is thought to repress transcription via methyl-CpG specific binding proteins, which recruit enzymatic machinery promoting the formation of inactive chromatin at targeted loci. Loss of DNA methylation can result in the activation of normally silent genes during mouse and amphibian development. Paradoxically, global changes in gene expression have not been observed in mice that are null for the methyl-CpG specific repressors MeCP2, MBD1 or MBD2. Here, we demonstrate that xKaiso, a novel methyl-CpG specific repressor protein, is required to maintain transcription silencing during early Xenopus laevis development. In the absence of xKaiso function, premature zygotic gene expression occurs before the mid-blastula transition (MBT). Subsequent phenotypes (developmental arrest and apoptosis) strongly resemble those observed for hypomethylated embryos. Injection of wild-type human kaiso mRNA can rescue the phenotype and associated gene expression changes of xKaiso-depleted embryos. Our results, including gene expression profiling, are consistent with an essential role for xKaiso as a global repressor of methylated genes during early vertebrate development. PMID- 15548583 TI - ENU induced mutations causing congenital cardiovascular anomalies. AB - We used non-invasive high frequency ultrasound to screen N-ethyl-N-nitrosourea mutagenized mouse fetuses for congenital cardiovascular anomalies. We ultrasound scanned 7546 mouse fetuses from 262 mutagenized families, and identified 124 families with cardiovascular defects. Represented were most of the major congenital cardiovascular anomalies seen clinically. The ENU-induced mutations in several families were mapped using polymorphic microsatellite DNA markers. One family with forelimb anomalies and ventricular septal defects, phenotypes similar to Holt-Oram syndrome, and one family with transposition of the great arteries and heart situs anomalies were mapped to different regions of mouse chromosome 4. A third mutation causing persistent truncus arteriosus and craniofacial defects, phenotypes reminiscent of DiGeorge syndrome, was mapped to mouse chromosome 2. We note that mouse chromosomes 4 and 2 do not contain Tbx5 or Tbx1, genes previously linked to Holt-Oram and DiGeorge syndromes, respectively. In two other families, the ENU-induced mutation was identified--Sema3CL605P was associated with persistent truncus arteriosus with interrupted aortic arch, and the Gja1W45X connexin43 mutation caused conotruncal malformation and coronary aneurysms. Although our screen was designed as a recessive screen, a number of the mutations showed cardiovascular phenotypes in both heterozygote and homozygote animals. These studies show the efficacy of ENU mutagenesis and high-throughput ultrasound phenotyping in recovering mutations causing a wide spectrum of congenital heart defects. These ENU-induced mutations hold promise in yielding new insights into the genetic basis for human congenital heart disease. PMID- 15548584 TI - Regional requirements for Dishevelled signaling during Xenopus gastrulation: separable effects on blastopore closure, mesendoderm internalization and archenteron formation. AB - During amphibian gastrulation, the embryo is transformed by the combined actions of several different tissues. Paradoxically, many of these morphogenetic processes can occur autonomously in tissue explants, yet the tissues in intact embryos must interact and be coordinated with one another in order to accomplish the major goals of gastrulation: closure of the blastopore to bring the endoderm and mesoderm fully inside the ectoderm, and generation of the archenteron. Here, we present high-resolution 3D digital datasets of frog gastrulae, and morphometrics that allow simultaneous assessment of the progress of convergent extension, blastopore closure and archenteron formation in a single embryo. To examine how the diverse morphogenetic engines work together to accomplish gastrulation, we combined these tools with time-lapse analysis of gastrulation, and examined both wild-type embryos and embryos in which gastrulation was disrupted by the manipulation of Dishevelled (Xdsh) signaling. Remarkably, although inhibition of Xdsh signaling disrupted both convergent extension and blastopore closure, mesendoderm internalization proceeded very effectively in these embryos. In addition, much of archenteron elongation was found to be independent of Xdsh signaling, especially during the second half of gastrulation. Finally, even in normal embryos, we found a surprising degree of dissociability between the various morphogenetic processes that occur during gastrulation. Together, these data highlight the central role of PCP signaling in governing distinct events of Xenopus gastrulation, and suggest that the loose relationship between morphogenetic processes may have facilitated the evolution of the wide variety of gastrulation mechanisms seen in different amphibian species. PMID- 15548585 TI - GDF11 modulates NGN3+ islet progenitor cell number and promotes beta-cell differentiation in pancreas development. AB - Identification of endogenous signals that regulate expansion and maturation of organ-specific progenitor cells is a major goal in studies of organ development. Here we provide evidence that growth differentiation factor 11 (GDF11), a member of the TGF-beta ligand family, governs the number and maturation of islet progenitor cells in mouse pancreas development. Gdf11 is expressed in embryonic pancreatic epithelium during formation of islet progenitor cells that express neurogenin 3. Mice deficient for Gdf11 harbor increased numbers of NGN3+ cells, revealing that GDF11 negatively regulates production of islet progenitor cells. Despite a marked expansion of these NGN3+ islet progenitors, mice lacking Gdf11 have reduced beta-cell numbers and evidence of arrested beta-cell development, indicating that GDF11 is also required for beta-cell maturation. Similar precursor and islet cell phenotypes are observed in mice deficient for SMAD2, an intracellular signaling factor activated by TGF-beta signals. Our data suggest that Gdf11 and Smad2 regulate islet cell differentiation in parallel to the Notch pathway, which previously has been shown to control development of NGN3+ cells. Thus, our studies reveal mechanisms by which GDF11 regulates the production and maturation of islet progenitor cells in pancreas development. PMID- 15548586 TI - In vivo evidence for short- and long-range cell communication in cranial neural crest cells. AB - The proper assembly of craniofacial structures and the peripheral nervous system requires neural crest cells to emerge from the neural tube and navigate over long distances to the branchial arches. Cell and molecular studies have shed light on potential intrinsic and extrinsic cues, which, in combination, are thought to ensure the induction and specification of cranial neural crest cells. However, much less is known about how migrating neural crest cells interpret and integrate signals from the microenvironment and other neural crest cells to sort into and maintain the stereotypical pattern of three spatially segregated streams. Here, we explore the extent to which cranial neural crest cells use cell-to-cell and cell-environment interactions to pathfind. The cell membrane and cytoskeletal elements in chick premigratory neural crest cells were labeled in vivo. Three dimensional reconstructions of migrating neural crest cells were then obtained using confocal static and time-lapse imaging. It was found that neural crest cells maintained nearly constant contact with other migrating neural crest cells, in addition to the microenvironment. Cells used lamellipodia or short, thin filopodia (1-2 microm wide) for local contacts (<20 microm). Non-local, long distance contact (up to 100 microm) was initiated by filopodia that extended and retracted, extended and tracked, or tethered two non-neighboring cells. Intriguingly, the cell-to-cell contacts often stimulated a cell to change direction in favor of a neighboring cell's trajectory. In summary, our results present in vivo evidence for local and long-range neural crest cell interactions, suggesting a possible role for these contacts in directional guidance. PMID- 15548587 TI - Theobromine inhibits sensory nerve activation and cough. AB - Cough is a common and protective reflex, but persistent coughing is debilitating and impairs quality of life. Antitussive treatment using opioids is limited by unacceptable side effects, and there is a great need for more effective remedies. The present study demonstrates that theobromine, a methylxanthine derivative present in cocoa, effectively inhibits citric acid-induced cough in guinea-pigs in vivo. Furthermore, in a randomized, double-blind, placebo-controlled study in man, theobromine suppresses capsaicin-induced cough with no adverse effects. We also demonstrate that theobromine directly inhibits capsaicin-induced sensory nerve depolarization of guinea-pig and human vagus nerve suggestive of an inhibitory effect on afferent nerve activation. These data indicate the actions of theobromine appear to be peripherally mediated. We conclude theobromine is a novel and promising treatment, which may form the basis for a new class of antitussive drugs. PMID- 15548588 TI - Molecular profiling of human prostate tissues: insights into gene expression patterns of prostate development during puberty. AB - Testosterone production surges during puberty and orchestrates massive growth and reorganization of the prostate gland, and this glandular architecture is maintained thereafter throughout adulthood. Benign prostatic hyperplasia (BPH) and prostate adenocarcinoma (PCA) are common diseases in adulthood that do not develop in the absence of androgens. Our objective was to gain insight into gene expression changes of the prostate gland at puberty, a crucial juncture in prostate development that is androgen dependent. Understanding the role played by androgens in normal prostate development may provide greater insight into androgen involvement in prostatic diseases. Benign prostate tissues obtained from pubertal and adult age group cadaveric organ donors were harvested and profiled using 20,000 element cDNA microarrays. Statistical analysis of the microarray data identified 375 genes that were differentially expressed in pubertal prostates relative to adult prostates including genes such as Nkx3.1, TMEPAI, TGFBR3, FASN, ANKH, TGFBR2, FAAH, S100P, HoxB13, fibronectin, and TSC2 among others. Comparisons of pubertal and BPH expression profiles revealed a subset of genes that shared the expression pattern between the two groups. In addition, we observed that several genes from this list were previously demonstrated to be regulated by androgen and hence could also be potential in vivo targets of androgen action in the pubertal human prostate. Promoter searches revealed the presence of androgen response elements in a cohort of genes including tumor necrosis factor-alpha induced adipose related protein, which was found to be induced by androgen. In summary, this is the first report that provides a comprehensive view of the molecular events that occur during puberty in the human prostate and provides a cohort of genes that could be potential in vivo targets of androgenic action during puberty. PMID- 15548589 TI - Intracellular Abeta42 activates p53 promoter: a pathway to neurodegeneration in Alzheimer's disease. AB - The amyloid beta-protein (Abeta) ending at 42 plays a pivotal role in Alzheimer's disease (AD). We have reported previously that intracellular Abeta42 is associated with neuronal apoptosis in vitro and in vivo. Here, we show that intracellular Abeta42 directly activated the p53 promoter, resulting in p53 dependent apoptosis, and that intracellular Abeta40 had a similar but lesser effect. Moreover, oxidative DNA damage induced nuclear localization of Abeta42 with p53 mRNA elevation in guinea-pig primary neurons. Also, p53 expression was elevated in brain of sporadic AD and transgenic mice carrying mutant familial AD genes. Remarkably, accumulation of both Abeta42 and p53 was found in some degenerating-shape neurons in both transgenic mice and human AD cases. Thus, the intracellular Abeta42/p53 pathway may be directly relevant to neuronal loss in AD. Although neurotoxicity of extracellular Abeta is well known and synaptic/mitochondrial dysfunction by intracellular Abeta42 has recently been suggested, intracellular Abeta42 may cause p53-dependent neuronal apoptosis through activation of the p53 promoter; thus demonstrating an alternative pathogenesis in AD. PMID- 15548590 TI - Munc13-4 is an effector of rab27a and controls secretion of lysosomes in hematopoietic cells. AB - Griscelli syndrome type 2 (GS2) is a genetic disorder in which patients exhibit life-threatening defects of cytotoxic T lymphocytes (CTLs) whose lytic granules fail to dock on the plasma membrane and therefore do not release their contents. The disease is caused by the absence of functional rab27a, but how rab27a controls secretion of lytic granule contents remains elusive. Mutations in Munc13 4 cause familial hemophagocytic lymphohistiocytosis subtype 3 (FHL3), a disease phenotypically related to GS2. We show that Munc13-4 is a direct partner of rab27a. The two proteins are highly expressed in CTLs and mast cells where they colocalize on secretory lysosomes. The region comprising the Munc13 homology domains is essential for the localization of Munc13-4 to secretory lysosomes. The GS2 mutant rab27aW73G strongly reduced binding to Munc13-4, whereas the FHL3 mutant Munc13-4Delta608-611 failed to bind rab27a. Overexpression of Munc13-4 enhanced degranulation of secretory lysosomes in mast cells, showing that it has a positive regulatory role in secretory lysosome fusion. We suggest that the secretion defects seen in GS2 and FHL3 have a common origin, and we propose that the rab27a/Munc13-4 complex is an essential regulator of secretory granule fusion with the plasma membrane in hematopoietic cells. Mutations in either of the two genes prevent formation of this complex and abolish secretion. PMID- 15548591 TI - Slipping or gripping? Fluorescent speckle microscopy in fish keratocytes reveals two different mechanisms for generating a retrograde flow of actin. AB - Fish keratocytes can generate rearward directed traction forces within front portions of the lamellipodium, suggesting that a retrograde flow of actin may also occur here but this was not detected by previous photoactivation experiments. To investigate the relationship between retrograde flow and traction force generation, we have transfected keratocytes with GFP-actin and used fluorescent speckle microscopy, to observe speckle flow. We detected a retrograde flow of actin within the leading lamellipodium that is inversely proportional to both protrusion rate and cell speed. To observe the effect of reducing contractility, we treated transfected cells with ML7, a potent inhibitor of myosin II. Surprisingly, ML7 treatment led to an increase in retrograde flow rate, together with a decrease in protrusion and cell speed, but only in rapidly moving cells. In slower moving cells, retrograde flow decreased, whereas protrusion rate and cell speed increased. These results suggest that there are two mechanisms for producing retrograde flow. One involves slippage between the cytoskeleton and adhesions, that decreases traction force production. The other involves slippage between adhesions and the substratum, which increases traction force production. We conclude that a biphasic relationship exists between retrograde actin flow and adhesiveness in moving keratocytes. PMID- 15548592 TI - Hec1 and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites. AB - A major goal in the study of vertebrate mitosis is to identify proteins that create the kinetochore-microtubule attachment site. Attachment sites within the kinetochore outer plate generate microtubule dependent forces for chromosome movement and regulate spindle checkpoint protein assembly at the kinetochore. The Ndc80 complex, comprised of Ndc80 (Hec1), Nuf2, Spc24, and Spc25, is essential for metaphase chromosome alignment and anaphase chromosome segregation. It has also been suggested to have roles in kinetochore microtubule formation, production of kinetochore tension, and the spindle checkpoint. Here we show that Nuf2 and Hec1 localize throughout the outer plate, and not the corona, of the vertebrate kinetochore. They are part of a stable "core" region whose assembly dynamics are distinct from other outer domain spindle checkpoint and motor proteins. Furthermore, Nuf2 and Hec1 are required for formation and/or maintenance of the outer plate structure itself. Fluorescence light microscopy, live cell imaging, and electron microscopy provide quantitative data demonstrating that Nuf2 and Hec1 are essential for normal kinetochore microtubule attachment. Our results indicate that Nuf2 and Hec1 are required for organization of stable microtubule plus-end binding sites in the outer plate that are needed for the sustained poleward forces required for biorientation at kinetochores. PMID- 15548594 TI - The role of microtubules in rapid hyphal tip growth of Aspergillus nidulans. AB - The filamentous fungus Aspergillus nidulans grows by polarized extension of hyphal tips. The actin cytoskeleton is essential for polarized growth, but the role of microtubules has been controversial. To define the role of microtubules in tip growth, we used time-lapse microscopy to measure tip growth rates in germlings of A. nidulans and in multinucleate hyphal tip cells, and we used a green fluorescent protein-alpha-tubulin fusion to observe the effects of the antimicrotubule agent benomyl. Hyphal tip cells grew approximately 5 times faster than binucleate germlings. In germlings, cytoplasmic microtubules disassembled completely in mitosis. In hyphal tip cells, however, microtubules disassembled through most of the cytoplasm in mitosis but persisted in a region near the hyphal tip. The growth rate of hyphal tip cells did not change significantly in mitosis. Benomyl caused rapid disassembly of microtubules in tip cells and a 10x reduction in growth rate. When benomyl was washed out, microtubules assembled quickly and rapid tip growth resumed. These results demonstrate that although microtubules are not strictly required for polarized growth, they are rate limiting for the growth of hyphal tip cells. These data also reveal that A. nidulans exhibits a remarkable spatial regulation of microtubule disassembly within hyphal tip cells. PMID- 15548593 TI - Fractionation of the epithelial apical junctional complex: reassessment of protein distributions in different substructures. AB - The epithelial apical junctional complex (AJC) is an important regulator of cell structure and function. The AJC is compartmentalized into substructures comprising the tight and adherens junctions, and other membrane complexes containing the membrane proteins nectin, junctional adhesion molecule, and crumbs. In addition, many peripheral membrane proteins localize to the AJC. Studies of isolated proteins indicate a complex map of potential binding partners in which there is extensive overlap in the interactions between proteins in different AJC substructures. As an alternative to a direct search for specific protein-protein interactions, we sought to separate membrane substructures of the AJC in iodixanol density gradients and define their protein constituents. Results show that the AJC can be fractured into membrane substructures that contain specific membrane and peripheral membrane proteins. The composition of each substructure reveals a more limited overlap in common proteins than predicted from the inventory of potential interactions; some of the overlapping proteins may be involved in stepwise recruitment and assembly of AJC substructures. PMID- 15548595 TI - Xrs2p regulates Mre11p translocation to the nucleus and plays a role in telomere elongation and meiotic recombination. AB - The Mre11-Rad50-Xrs2 (MRX) protein complex plays pivotal roles in meiotic recombination, repair of damaged DNA, telomere elongation, and cell cycle checkpoint control. Xrs2p is known to be essential for all the functions of the complex, but its role in the complex has not been clearly elucidated. A 32-amino acid region near the C terminus of Xrs2p was identified as an Mre11p-binding site. No more function of Xrs2p than translocation of Mre11p from the cytoplasm to the nucleus is necessary for response to DNA damage. However, domains in Xrs2p located both 49 amino acids upstream and 104 amino acids downstream of the Mre11p binding site are required for meiotic recombination and telomere elongation, respectively, in addition to the 32-amino acid region. These findings demonstrate that Xrs2p acts as a specificity factor that allows the MRX complex to function in meiotic recombination and in telomere elongation. PMID- 15548596 TI - Analysis of mutant phenotypes and splicing defects demonstrates functional collaboration between the large and small subunits of the essential splicing factor U2AF in vivo. AB - The heterodimeric splicing factor U2AF plays an important role in 3' splice site selection, but the division of labor between the two subunits in vivo remains unclear. In vitro assays led to the proposal that the human large subunit recognizes 3' splice sites with extensive polypyrimidine tracts independently of the small subunit. We report in vivo analysis demonstrating that all five domains of spU2AFLG are essential for viability; a partial deletion of the linker region, which forms the small subunit interface, produces a severe growth defect and an aberrant morphology. A small subunit zinc-binding domain mutant confers a similar phenotype, suggesting that the heterodimer functions as a unit during splicing in Schizosaccharomyces pombe. As this is not predicted by the model for metazoan 3' splice site recognition, we sought introns for which the spU2AFLG and spU2AFSM make distinct contributions by analyzing diverse splicing events in strains harboring mutations in each partner. Requirements for the two subunits are generally parallel and, moreover, do not correlate with the length or strength of the 3' pyrimidine tract. These and other studies performed in fission yeast support a model for 3' splice site recognition in which the two subunits of U2AF functionally collaborate in vivo. PMID- 15548597 TI - The Caenorhabditis elegans Aurora B kinase AIR-2 phosphorylates and is required for the localization of a BimC kinesin to meiotic and mitotic spindles. AB - BimC kinesins are required for mitotic spindle assembly in a variety of organisms. These proteins are localized to centrosomes, spindle microtubules, and the spindle midzone. We have previously shown that the Caenorhabditis elegans Aurora B kinase AIR-2 is required for the localization of the ZEN-4 kinesin protein to midzone microtubules. To determine whether the association of BimC kinesins with spindle microtubules is also dependent on AIR-2, we examined the expression pattern of BMK-1, a C. elegans BimC kinesin, in wild-type and AIR-2 deficient embryos. BMK-1 is highly expressed in the hermaphrodite gonad and is localized to meiotic spindle microtubules in the newly fertilized embryo. In mitotic embryos, BMK-1 is associated with spindle microtubules from prophase through anaphase and is concentrated at the spindle midzone during anaphase and telophase. In the absence of AIR-2, BMK-1 localization to meiotic and mitotic spindles is greatly reduced. This is not a consequence of loss of ZEN-4 localization because BMK-1 is appropriately localized in ZEN-4-deficient embryos. Furthermore, AIR-2 and BMK-1 directly interact with one another and the C terminal tail domain of BMK-1 is specifically phosphorylated by AIR-2 in vitro. Together with our previous data, these results suggest that at least one function of the Aurora B kinases is to recruit spindle-associated motor proteins to their sites of action. PMID- 15548598 TI - HGF converts ErbB2/Neu epithelial morphogenesis to cell invasion. AB - Activation of the hepatocyte growth factor receptor Met induces a morphogenic response and stimulates the formation of branching tubules by Madin-Darby canine kidney (MDCK) epithelial cells in three-dimensional cultures. A constitutively activated ErbB2/Neu receptor, NeuNT, promotes a similar invasive morphogenic program in MDCK cells. Because both receptors are expressed in breast epithelia, are associated with poor prognosis, and hepatocyte growth factor (HGF) is expressed in stroma, we examined the consequence of cooperation between these signals. We show that HGF disrupts NeuNT-induced epithelial morphogenesis, stimulating the breakdown of cell-cell junctions, dispersal, and invasion of single cells. This correlates with a decrease in junctional proteins claudin-1 and E-cadherin, in addition to the internalization of the tight junction protein ZO-1. HGF-induced invasion of NT-expressing cells is abrogated by pretreatment with a pharmacological inhibitor of the mitogen-activated protein kinase kinase (MEK) pathway, which restores E-cadherin and ZO-1 at cell-cell junctions, establishing the involvement of MEK-dependent pathways in this process. These results demonstrate that physiological signals downstream from the HGF/Met receptor synergize with ErbB2/Neu to enhance the malignant phenotype, promoting the breakdown of cell-cell junctions and enhanced cell invasion. This is particularly important for cancers where ErbB2/Neu is overexpressed and HGF is a physiological growth factor found in the stroma. PMID- 15548599 TI - Actin-depolymerizing factor and cofilin-1 play overlapping roles in promoting rapid F-actin depolymerization in mammalian nonmuscle cells. AB - Actin-depolymerizing factor (ADF)/cofilins are small actin-binding proteins found in all eukaryotes. In vitro, ADF/cofilins promote actin dynamics by depolymerizing and severing actin filaments. However, whether ADF/cofilins contribute to actin dynamics in cells by disassembling "old" actin filaments or by promoting actin filament assembly through their severing activity is a matter of controversy. Analysis of mammalian ADF/cofilins is further complicated by the presence of multiple isoforms, which may contribute to actin dynamics by different mechanisms. We show that two isoforms, ADF and cofilin-1, are expressed in mouse NIH 3T3, B16F1, and Neuro 2A cells. Depleting cofilin-1 and/or ADF by siRNA leads to an accumulation of F-actin and to an increase in cell size. Cofilin-1 and ADF seem to play overlapping roles in cells, because the knockdown phenotype of either protein could be rescued by overexpression of the other one. Cofilin-1 and ADF knockdown cells also had defects in cell motility and cytokinesis, and these defects were most pronounced when both ADF and cofilin-1 were depleted. Fluorescence recovery after photobleaching analysis and studies with an actin monomer-sequestering drug, latrunculin-A, demonstrated that these phenotypes arose from diminished actin filament depolymerization rates. These data suggest that mammalian ADF and cofilin-1 promote cytoskeletal dynamics by depolymerizing actin filaments and that this activity is critical for several processes such as cytokinesis and cell motility. PMID- 15548600 TI - Identification and characterization of regulator of G protein signaling 4 (RGS4) as a novel inhibitor of tubulogenesis: RGS4 inhibits mitogen-activated protein kinases and vascular endothelial growth factor signaling. AB - Tubulogenesis by epithelial cells regulates kidney, lung, and mammary development, whereas that by endothelial cells regulates vascular development. Although functionally dissimilar, the processes necessary for tubulation by epithelial and endothelial cells are very similar. We performed microarray analysis to further our understanding of tubulogenesis and observed a robust induction of regulator of G protein signaling 4 (RGS4) mRNA expression solely in tubulating cells, thereby implicating RGS4 as a potential regulator of tubulogenesis. Accordingly, RGS4 overexpression delayed and altered lung epithelial cell tubulation by selectively inhibiting G protein-mediated p38 MAPK activation, and, consequently, by reducing epithelial cell proliferation, migration, and expression of vascular endothelial growth factor (VEGF). The tubulogenic defects imparted by RGS4 in epithelial cells, including its reduction in VEGF expression, were rescued by overexpression of constitutively active MKK6, an activator of p38 MAPK. Similarly, RGS4 overexpression abrogated endothelial cell angiogenic sprouting by inhibiting their synthesis of DNA and invasion through synthetic basement membranes. We further show that RGS4 expression antagonized VEGF stimulation of DNA synthesis and extracellular signal-regulated kinase (ERK)1/ERK2 and p38 MAPK activation as well as ERK1/ERK2 activation stimulated by endothelin-1 and angiotensin II. RGS4 had no effect on the phosphorylation of Smad1 and Smad2 by bone morphogenic protein-7 and transforming growth factor-beta, respectively, indicating that RGS4 selectively inhibits G protein and VEGF signaling in endothelial cells. Finally, we found that RGS4 reduced endothelial cell response to VEGF by decreasing VEGF receptor-2 (KDR) expression. We therefore propose RGS4 as a novel antagonist of epithelial and endothelial cell tubulogenesis that selectively antagonizes intracellular signaling by G proteins and VEGF, thereby inhibiting cell proliferation, migration, and invasion, and VEGF and KDR expression. PMID- 15548601 TI - The Arabidopsis peroxisomal targeting signal type 2 receptor PEX7 is necessary for peroxisome function and dependent on PEX5. AB - Plant peroxisomal proteins catalyze key metabolic reactions. Several peroxisome biogenesis PEROXIN (PEX) genes encode proteins acting in the import of targeted proteins necessary for these processes into the peroxisomal matrix. Most peroxisomal matrix proteins bear characterized Peroxisomal Targeting Signals (PTS1 or PTS2), which are bound by the receptors PEX5 or PEX7, respectively, for import into peroxisomes. Here we describe the isolation and characterization of an Arabidopsis peroxin mutant, pex7-1, which displays peroxisome-defective phenotypes including reduced PTS2 protein import. We also demonstrate that the pex5-1 PTS1 receptor mutant, which contains a lesion in a domain conserved among PEX7-binding proteins from various organisms, is defective not in PTS1 protein import, but rather in PTS2 protein import. Combining these mutations in a pex7-1 pex5-1 double mutant abolishes detectable PTS2 protein import and yields seedlings that are entirely sucrose-dependent for establishment, suggesting a severe block in peroxisomal fatty acid beta-oxidation. Adult pex7-1 pex5-1 plants have reduced stature and bear abnormally shaped seeds, few of which are viable. The pex7-1 pex5-1 seedlings that germinate have dramatically fewer lateral roots and often display fused cotyledons, phenotypes associated with reduced auxin response. Thus PTS2-directed peroxisomal import is necessary for normal embryonic development, seedling establishment, and vegetative growth. PMID- 15548602 TI - Cdc42 mediates nucleus movement and MTOC polarization in Swiss 3T3 fibroblasts under mechanical shear stress. AB - Nucleus movement is essential during nucleus positioning for tissue growth and development in eukaryotic cells. However, molecular regulators of nucleus movement in interphase fibroblasts have yet to be identified. Here, we report that nuclei of Swiss 3T3 fibroblasts undergo enhanced movement when subjected to shear flows. Such movement includes both rotation and translocation and is dependent on microtubule, not F-actin, structure. Through inactivation of Rho GTPases, well-known mediators of cytoskeleton reorganization, we demonstrate that Cdc42, not RhoA or Rac1, controls the extent of nucleus translocation, and more importantly, of nucleus rotation in the cytoplasm. In addition to generating nuclei movement, we find that shear flows also causes repositioning of the MTOC in the direction of flow. This behavior is also controlled by Cdc42 via the Par6/protein kinase Czeta pathway. These results are the first to establish Cdc42 as a molecular regulator of not only shear-induced MTOC polarization in Swiss 3T3 fibroblasts, but also of shear-induced microtubule-dependent nucleus movement. We propose that the movements of MTOC and nucleus are coupled chemically, because they are both regulated by Cdc42 and dependent on microtubule structure, and physically, possibly via Hook/SUN family homologues similar to those found in Caenorhabditis elegans. PMID- 15548603 TI - Changes in genomewide occupancy of core transcriptional regulators during heat stress. AB - Organisms respond to heat stress by reprogramming gene expression. In Saccharomyces cerevisiae, heat-induced genes tend to be regulated by factors that belong to the Spt-Ada-Gcn5 acetyltransferase (SAGA) transcription regulatory pathway, whereas heat-repressed genes tend to be regulated by a parallel pathway involving transcription factor IID (TFIID). Here, we examine whether heat stress affects the occupancy of representative factors of each pathway at promoter regions throughout the yeast genome. Representatives of the SAGA pathway include the TATA binding protein, Spt3, and Mot1. Representatives of the TFIID pathway include the TATA binding protein, TAF1, and Bdf1. We find that heat stress causes disassembly of the TFIID pathway at genes that are inhibited by stress. In contrast, heat induces assembly of the SAGA pathway at stress-induced genes, although many also assemble along the TFIID pathway. Other genes were found to assemble almost exclusively along the TFIID pathway. Strikingly, these genes are lowly transcribed and are generally not induced. Thus, heat stress leads to factor assembly along each pathway but with distinct transcriptional outcomes. Further investigation of these pathways reveals that Bdf1 and Mot1 negatively regulate the SAGA pathway in different ways. The findings suggest that Bdf1 blocks assembly, whereas Mot1 promotes disassembly of the transcription machinery. PMID- 15548604 TI - A gene encoding a putative FAD-dependent L-2-hydroxyglutarate dehydrogenase is mutated in L-2-hydroxyglutaric aciduria. AB - The purpose of this study was to identify the biochemical and genetic defect in L 2-hydroxyglutaric aciduria, a neurometabolic disorder characterized by the presence of elevated concentrations of L-2-hydroxyglutaric acid in urine, plasma, and cerebrospinal fluid. Evidence is provided for the existence in rat tissues of a FAD-dependent enzyme catalyzing specifically the oxidation of L-2 hydroxyglutarate to alpha-ketoglutarate. This enzyme is mainly expressed in liver and kidney but also at lower levels in heart, brain, and other tissues. Subcellular fractionation indicates that the liver enzyme is present in mitochondria, where it is bound to membranes. Based on this information, a database search led to the identification of a gene encoding a human hypothetical protein homologous to bacterial FAD-dependent malate dehydrogenases and targeted to mitochondria. The gene encoding this protein, present on chromosome 14q22.1, was found to be in a region homozygous in patients with L-2-hydroxyglutaric aciduria from two consanguineous families. Three mutations that replaced a highly conserved residue (Lys-71-Glu and Glu-176-Asp) or removed exon 9 were identified in homozygous state in patients from three distinct families and were found to cosegregate with the disease. It is concluded that L-2-hydroxyglutarate is normally metabolized to alpha-ketoglutarate in mammalian tissues and that L-2 hydroxyglutaric aciduria is caused by mutations in the gene that most likely encodes L-2-hydroxyglutarate dehydrogenase. The pathological findings observed in this metabolic disorder must therefore be due to a toxic effect of L-2 hydroxyglutarate on the central nervous system. PMID- 15548605 TI - Compound library development guided by protein structure similarity clustering and natural product structure. AB - To identify biologically relevant and drug-like protein ligands for medicinal chemistry and chemical biology research the grouping of proteins according to evolutionary relationships and conservation of molecular recognition is an established method. We propose to employ structure similarity clustering of the ligand-sensing cores of protein domains (PSSC) in conjunction with natural product guided compound library development as a synergistic approach for the identification of biologically prevalidated ligands with high fidelity. This is supported by the concepts that (i) in nature spatial structure is more conserved than amino acid sequence, (ii) the number of fold types characteristic for all protein domains is limited, and (iii) the underlying frameworks of natural product classes with multiple biological activities provide evolutionarily selected starting points in structural space. On the basis of domain core similarity considerations and irrespective of sequence similarity, Cdc25A phosphatase, acetylcholinesterase, and 11beta-hydroxysteroid dehydrogenases type 1 and type 2 were grouped into a similarity cluster. A 147-member compound collection derived from the naturally occurring Cdc25A inhibitor dysidiolide yielded potent and selective inhibitors of the other members of the similarity cluster with a hit rate of 2-3%. Protein structure similarity clustering may provide an experimental opportunity to identify supersites in proteins. PMID- 15548606 TI - Linkage analysis of ordinal traits for pedigree data. AB - Linkage analysis is used routinely to map genes for human diseases and conditions. However, the existing linkage-analysis methods require that the diseases or conditions either be dichotomized or measured by a quantitative trait, such as blood pressure for hypertension. In the latter case, normality is generally assumed for the trait. However, many diseases and conditions, such as cancer and mental and behavioral conditions, are rated on ordinal scales. The objective of this study was to establish a framework to conduct linkage analysis for ordinal traits. We propose a latent-variable, proportional-odds logistic model that relates inheritance patterns to the distribution of the ordinal trait. We use the likelihood-ratio test for testing evidence of linkage. By means of simulation studies, we find that the power of our proposed model is substantially higher than that of the binary-trait-based linkage analysis and that our test statistic is robust with regard to certain parameter misspecifications. By using our proposed method, we performed a genome scan of the hoarding phenotype in a data set with 53 nuclear families, which were collected by the Tourette Syndrome Association International Consortium for Genetics (TSAICG). Standard linkage scans using hoarding as a dichotomous trait were also performed by using GENEHUNTER and ALLEGRO. Both GENEHUNTER and ALLEGRO failed to reveal any marker significantly linked to the binary hoarding phenotypes. However, our method identified three markers at 4q34-35 (P = 0.0009), 5q35.2-35.3 (P = 0.0001), and 17q25 (P = 0.0005) that manifest significant allele sharing. PMID- 15548607 TI - Transplanted human bone marrow contributes to vascular endothelium. AB - Recent evidence indicates that bone marrow is a source of endothelial progenitor cells that are mobilized into the peripheral blood in response to cytokines or tissue injury. Previously, we showed that functional endothelial cells (ECs) can be clonally derived from phenotypically defined hematopoietic stem cells. To determine the EC potential of human bone marrow and peripheral blood stem cells, blood vessels in sex-mismatched transplant recipients were evaluated. EC outcomes were identified by using a combination of immunohistochemistry and XY interphase FISH. Donor-derived ECs were detected in the skin and gut of transplant recipients with a mean frequency of 2% and could readily be distinguished from CD45-expressing hematopoietic stem cells. None of the >4,000 ECs examined had more than two sex chromosomes, consistent with an absence of cell fusion. Y chromosome signals were not detected in sex-matched female recipients, excluding the vertical transmission of male cells. None of the recipients evaluated before hematopoietic engraftment demonstrated donor-derived ECs, indicating a close linkage between the recovery of hematopoiesis and EC outcomes. Transplantable bone marrow-derived endothelial progenitor cells may represent novel therapeutic targets for hematopoietic and vascular disease. PMID- 15548608 TI - Function recovery after chemobleaching (FRAC): evidence for activity silent membrane receptors on cell surface. AB - Membrane proteins represent approximately 30% of the proteome of both prokaryotes and eukaryotes. Unique to cell surface receptors is their biogenesis pathway, which involves vesicular trafficking from the endoplasmic reticulum through the Golgi apparatus and to the cell surface. Increasing evidence suggests specific regulation of biogenesis for different membrane receptors, hence affecting their surface expression. We report the development of a pulse-chase assay to monitor function recovery after chemobleaching (FRAC) to probe the transit time of the Kir2.1 K+ channel to reach the cell surface. Our results reveal that the channel activity is contributed by a small fraction of channel protein, providing evidence of activity-silent "sleeping" molecules on the cell surface. This method distinguishes molecular density from functional density, and the assay strategy is generally applicable to other membrane receptors. The ability of the reported method to access the biogenesis pathways in a high-throughput manner facilitates the identification and evaluation of molecules affecting receptor trafficking. PMID- 15548609 TI - A small-molecule switch for Golgi sulfotransferases. AB - The study of glycan function is a major frontier in biology that could benefit from small molecules capable of perturbing carbohydrate structures on cells. The widespread role of sulfotransferases in modulating glycan function makes them prime targets for small-molecule modulators. Here, we report a system for conditional activation of Golgi-resident sulfotransferases using a chemical inducer of dimerization. Our approach capitalizes on two features shared by these enzymes: their requirement of Golgi localization for activity on cellular substrates and the modularity of their catalytic and localization domains. Fusion of these domains to the proteins FRB and FKBP enabled their induced assembly by the natural product rapamycin. We applied this strategy to the GlcNAc-6 sulfotransferases GlcNAc6ST-1 and GlcNAc6ST-2, which collaborate in the sulfation of L-selectin ligands. Both the activity and specificity of the inducible enzymes were indistinguishable from their WT counterparts. We further generated rapamycin inducible chimeric enzymes comprising the localization domain of a sulfotransferase and the catalytic domain of a glycosyltransferase, demonstrating the generality of the system among other Golgi enzymes. The approach provides a means for studying sulfate-dependent processes in cellular systems and, potentially, in vivo. PMID- 15548610 TI - GC/AT-content spikes as genomic punctuation marks. AB - Large-scale analysis of the GC-content distribution at the gene level reveals both common features and basic differences in genomes of different groups of species. Sharp changes in GC content are detected at the transcription boundaries for all species analyzed, including human, mouse, rat, chicken, fruit fly, and worm. However, two substantially distinct groups of GC-content profiles can be recognized: warm-blooded vertebrates including human, mouse, rat, and chicken, and invertebrates including fruit fly and worm. In vertebrates, sharp positive and negative spikes of GC content are observed at the transcription start and stop sites, respectively, and there is also a progressive decrease in GC content from the 5' untranslated region to the 3' untranslated region along the gene. In invertebrates, the positive and negative GC-content spikes at the transcription start and stop sites are preceded by spikes of opposite value, and the highest GC content is found in the coding regions of the genes. Cross-correlation analysis indicates high frequencies of GC-content spikes at transcription start and stop sites. The strong conservation of this genomic feature seen in comparisons of the human/mouse and human/rat orthologs, and the clustering of genes with GC-content spikes on chromosomes imply a biological function. The GC-content spikes at transcription boundaries may reflect a general principle of genomic punctuation. Our analysis also provides means for identifying these GC-content spikes in individual genomic sequences. PMID- 15548611 TI - Bv8 and endocrine gland-derived vascular endothelial growth factor stimulate hematopoiesis and hematopoietic cell mobilization. AB - Bv8 and endocrine-gland-derived VEGF (EG-VEGF), or prokineticins, are two highly related, secreted proteins that we previously described as selective angiogenic mitogens. Here we describe the expression and functional characterization of Bv8 in peripheral blood cells, notably monocytes, neutrophils, and dendritic cells, and in the bone marrow. In human and mouse, the two Bv8 G protein-coupled receptors are expressed in hematopoietic stem cells and specific mature blood cells, including lymphocytes. Bv8 is highly expressed by neutrophils at sites of inflammation and can stimulate migration of monocytes, in a pertussis toxin sensitive manner. Bv8, or EG-VEGF that shares the same receptors, increased numbers of colony-forming units granulocytic and monocytic in cultures of human or mouse hematopoietic stem cells. Systemic in vivo exposure to Bv8 or EG-VEGF resulted in significant increases in total leukocyte, neutrophil, and monocyte counts. Additionally, adenovirus (Av)Bv8 or AvEG-VEGF delivered just before 5 fluorouracil injury promoted the survival of hematopoietic cells and enhanced progenitor mobilization. In conclusion, Bv8 can promote survival and differentiation of the granulocytic and monocytic lineages. Bv8 potentially modulates growth, survival, and function of cells of the innate and adaptive immune systems, possibly through autocrine or paracrine signaling mechanisms. PMID- 15548612 TI - Harnessing phytochrome's glowing potential. AB - Directed evolution of a cyanobacterial phytochrome was undertaken to elucidate the structural basis of its light sensory activity by remodeling the chemical environment of its linear tetrapyrrole prosthetic group. In addition to identifying a small region of the apoprotein critical for maintaining phytochrome's native spectroscopic properties, our studies revealed a tyrosine-to histidine mutation that transformed phytochrome into an intensely red fluorescent biliprotein. This tyrosine is conserved in all members of the phytochrome superfamily, implicating direct participation in the primary photoprocess of phytochromes. Fluorescent phytochrome mutants also hold great promise to expand the present repertoire of genetically encoded fluorescent proteins into the near infrared. PMID- 15548613 TI - The structure of carbonmonoxy neuroglobin reveals a heme-sliding mechanism for control of ligand affinity. AB - Neuroglobin (Ngb), a globular heme protein expressed in the brain of vertebrates, binds oxygen reversibly, with an affinity comparable to myoglobin (Mb). Despite low sequence identity, the overall 3D fold of Ngb and Mb is very similar. Unlike in Mb, in Ngb the sixth coordination position of the heme iron is occupied by the distal histidine, in the absence of an exogenous ligand. Endogenous ligation has been proposed as a unique mechanism for affinity regulation and ligand discrimination in heme proteins. This peculiarity might be related to the still unknown physiological function of Ngb. Here, we present the x-ray structure of CO bound ferrous murine Ngb at 1.7 A and a comparison with the 1.5-A structure of ferric bis-histidine Ngb. We have also used Fourier transform IR spectroscopy of WT and mutant CO-ligated Ngb to examine structural heterogeneity in the active site. Upon CO binding, the distal histidine retains (by and large) its position, whereas the heme group slides deeper into a preformed crevice, thereby reshaping the large cavity ( approximately 290 A(3)) connecting the distal and proximal heme sides with the bulk. The heme relocation is accompanied by a significant decrease of structural disorder, especially of the EF loop, which may be the signal whereby Ngb communicates hypoxic conditions. This unexpected structural change unveils a heme-sliding mechanism of affinity control that may be of significance to understanding Ngb's role in the pathophysiology of the brain. PMID- 15548614 TI - Fragile X mental retardation protein is necessary for neurotransmitter-activated protein translation at synapses. AB - Fragile X mental retardation is caused by absence of the RNA-binding protein fragile X mental retardation protein (FMRP), encoded by the FMR1 gene. There is increasing evidence that FMRP regulates transport and modulates translation of some mRNAs. We studied neurotransmitter-activated synaptic protein synthesis in fmr1-knockout mice. Synaptoneurosomes from knockout mice did not manifest accelerated polyribosome assembly or protein synthesis as it occurs in wild-type mice upon stimulation of group I metabotropic glutamate receptors. Direct activation of protein kinase C did not compensate in the knockout mouse, indicating that the FMRP-dependent step is further along the signaling pathway. Visual cortices of young knockout mice exhibited a lower proportion of dendritic spine synapses containing polyribosomes than did the cortices of wild-type mice, corroborating this finding in vivo. This deficit in rapid neurotransmitter controlled local translation of specific proteins may contribute to morphological and functional abnormalities observed in patients with fragile X syndrome. PMID- 15548615 TI - The molecular program induced in T cells undergoing homeostatic proliferation. AB - Naive T cells proliferate independently of cognate antigen when introduced into lymphopenic hosts. Lymphopenia-induced proliferation depends on low-affinity MHC/self-peptide complexes and on IL-7. To elucidate the intracellular signals mediating this proliferation, we analyzed changes in gene expression in naive CD8+ T cells at different times after their transfer into a lymphopenic environment. The genes induced in response to lymphopenia were largely an attenuated subset of those turned up by full antigenic stimulation, including genes related to cell cycling, whereas excluding genes specifically associated with effector activity. After the initial phase of proliferation in an empty compartment, the naive T cells adopted a stable pattern of gene expression similar to that of antigen-experienced memory cells. Thus, T cells proliferating in lymphopenic hosts do not exhibit a unique gene-expression profile, instead relying on "traditional" signals for this antigen-independent proliferation; this process ultimately results in differentiation to "authentic" memory cells. PMID- 15548616 TI - Evidence that translocation of anthrax toxin's lethal factor is initiated by entry of its N terminus into the protective antigen channel. AB - Entry of the enzymatic components of anthrax toxin [lethal factor (LF) and edema factor] into the cytosol of mammalian cells depends on the ability of the activated protective antigen (PA63) component to form a channel (pore) in the membrane of an acidic intracellular compartment. To investigate the mechanism of translocation, we characterized N-terminally truncated forms of the PA63-binding domain of LF (LFN). Deleting 27 or 36 residues strongly inhibited acid-triggered translocation of LFN across the plasma membrane of CHO-K1 cells and ablated the protein's ability to block PA63 channels in planar lipid bilayers at a small positive voltage (+20 mV). Fusing a H6-tag to the N terminus of the truncated proteins restored both translocation and channel-blocking activities. At +20 mV, N-terminal H6 and biotin tags were accessible to Ni2+ and streptavidin, respectively, added to the trans compartment of a planar bilayer. On the basis of these findings, we propose that the N terminus of PA63-bound LF or edema factor enters the PA63-channel under the influence of acidic pH and a positive transmembrane potential and initiates translocation in an N- to C-terminal direction. PMID- 15548617 TI - Recycling Frank: Spontaneous emergence of homochirality in noncatalytic systems. AB - In this work, we introduce a prebiotically relevant protometabolic pattern corresponding to an engine of deracemization by using an external energy source. The spontaneous formation of a nonracemic mixture of chiral compounds can be observed in out-of-equilibrium systems via a symmetry-breaking phenomenon. This observation is possible thanks to chirally selective autocatalytic reactions (Frank's model) [Frank, F. C. (1953) Biochim. Biophys. Acta 11, 459-463]. We show that the use of a Frank-like model in a recycled system composed of reversible chemical reactions, rather than the classical irreversible system, allows for the emergence of a synergetic autoinduction from simple reactions, without any autocatalytic or even catalytic reaction. This model is described as a theoretical framework, based on the stereoselective reactivity of preexisting chiral monomeric building blocks (polymerization, epimerization, and depolymerization) maintained out of equilibrium by a continuous energy income, via an activation reaction. It permits the self-conversion of all monomeric subunits into a single chiral configuration. Real prebiotic systems of amino acid derivatives can be described on this basis. They are shown to be able to spontaneously reach a stable nonracemic state in a few centuries. In such systems, the presence of epimerization reactions is no more destructive, but in contrast is the central driving force of the unstabilization of the racemic state. PMID- 15548618 TI - Properties of dopamine release and uptake in the songbird basal ganglia. AB - Vocal learning in songbirds requires a basal ganglia circuit termed the anterior forebrain pathway (AFP). The AFP is not required for song production, and its role in song learning is not well understood. Like the mammalian striatum, the striatal component of the AFP, Area X, receives dense dopaminergic innervation from the midbrain. Since dopamine (DA) clearly plays a crucial role in basal ganglia-mediated motor control and learning in mammals, it seems likely that DA signaling contributes importantly to the functions of Area X as well. In this study, we used voltammetric methods to detect subsecond changes in extracellular DA concentration to gain better understanding of the properties and regulation of DA release and uptake in Area X. We electrically stimulated Ca(2+)- and action potential-dependent release of an electroactive substance in Area X brain slices and identified the substance as DA by the voltammetric waveform, electrode selectivity, and neurochemical and pharmacological evidence. As in the mammalian striatum, DA release in Area X is depressed by autoinhibition, and the lifetime of extracellular DA is strongly constrained by monoamine transporters. These results add to the known physiological similarities of the mammalian and songbird striatum and support further use of voltammetry in songbirds to investigate the role of basal ganglia DA in motor learning. PMID- 15548619 TI - Plateau potentials in developing antennal-lobe neurons of the moth, Manduca sexta. AB - Using whole cell recordings from antennal-lobe (AL) neurons in vitro and in situ, in semi-intact brain preparations, we examined membrane properties that contribute to electrical activity exhibited by developing neurons in primary olfactory centers of the brain of the sphinx moth, Manduca sexta. This activity is characterized by prolonged periods of membrane depolarization that resemble plateau potentials. The presence of plateau potential-generating mechanisms was confirmed using a series of tests established earlier. Brief depolarizing current pulses could be used to trigger a plateau state. Once triggered, plateau potentials could be terminated by brief pulses of hyperpolarizing current. Both triggering and terminating of firing states were threshold phenomena, and both conditions resulted in all-or-none responses. Rebound excitation from prolonged hyperpolarizing pulses could also be used to generate plateau potentials in some cells. These neurons were found to express a hyperpolarization-activated inward current. Neither the generation nor the maintenance of plateau potentials was affected by removal of Na+ ions from the extracellular medium or by blockade of Na+ currents with TTX. However, blocking of Ca2+ currents with Cd2+ (5 x 10(-4) M) inhibited the generation of plateau potentials, indicating that, in Manduca AL neurons, plateau potentials depend on Ca2+. Examining Ca2+ currents in isolation revealed that activation of these currents occurs in the absence of experimentally applied depolarizing stimuli. Our results suggest that this activity underlies the generation of plateau potentials and characteristic bursts of electrical activity in developing AL neurons of M. sexta. PMID- 15548620 TI - Polytrodes: high-density silicon electrode arrays for large-scale multiunit recording. AB - We developed a variety of 54-channel high-density silicon electrode arrays (polytrodes) designed to record from large numbers of neurons spanning millimeters of brain. In cat visual cortex, it was possible to make simultaneous recordings from >100 well-isolated neurons. Using standard clustering methods, polytrodes provide a quality of single-unit isolation that surpasses that attainable with tetrodes. Guidelines for successful in vivo recording and precise electrode positioning are described. We also describe a high-bandwidth continuous data-acquisition system designed specifically for polytrodes and an automated impedance meter for testing polytrode site integrity. Despite having smaller interconnect pitches than earlier silicon-based electrodes of this type, these polytrodes have negligible channel crosstalk, comparable reliability, and low site impedances and are capable of making high-fidelity multiunit recordings with minimal tissue damage. The relatively benign nature of planar electrode arrays is evident both histologically and in experiments where the polytrode was repeatedly advanced and retracted hundreds of microns over periods of many hours. It was possible to maintain stable recordings from active neurons adjacent to the polytrode without change in their absolute positions, neurophysiological or receptive field properties. PMID- 15548621 TI - Effects of temperature on calcium transients and Ca2+-dependent afterhyperpolarizations in neocortical pyramidal neurons. AB - In neocortical pyramidal neurons, the medium (mAHP) and slow AHP (sAHP) have different relationships with intracellular [Ca2+]. To further explore these differences, we varied bath temperature and compared passive and active membrane properties and Ca2+ transients in response to a single action potential (AP) or trains of APs. We tested whether Ca(2+)-dependent events are more temperature sensitive than voltage-dependent ones, the slow rise time of the sAHP is limited by diffusion, and temperature sensitivity differs between the mAHP and sAHP. The onset and decay kinetics of the sAHP were very temperature sensitive (more so than diffusion). We found that the decay time course of Ca2+ transients was also very temperature sensitive. In contrast, the mAHP (amplitude, time to peak, and exponential decay) and sAHP peak amplitude were moderately sensitive to temperature. The amplitudes of intracellular Ca2+ transients evoked either by a single spike or a train of spikes showed modest temperature sensitivities. Pyramidal neuron input resistance was increased by cooling. With the exception of threshold, which remained unchanged between 22 and 35 degrees C, action potential parameters (amplitude, half-width, maximum rates of rise and fall) were modestly affected by temperature. Collectively, these data suggest that temperature sensitivity was higher for the Ca(2+)-dependent sAHP than for voltage-dependent AP parameters or for the mAHP, diffusion of Ca2+ over distance cannot explain the slow rise of the sAHP in these cells, and the kinetics of the sAHP and mAHP are affected differently by temperature. PMID- 15548622 TI - Spatial distribution of inputs and local receptive field properties of a wide field, looming sensitive neuron. AB - The lobula giant movement detector (LGMD) in the locust visual system and its target neuron, the descending contralateral movement detector (DCMD), respond to approaching objects looming on a collision course with the animal. They thus provide a good model to study the cellular and network mechanisms underlying the sensitivity to this specific class of behaviorally relevant stimuli. We determined over an entire locust eye the density distribution of optical axes describing the spatial organization of local inputs to the visual system and compared it with the sensitivity distribution of the LGMD/DCMD to local motion stimuli. The density of optical axes peaks in the equatorial region of the frontal eye. Local motion sensitivity, however, peaks in the equatorial region of the caudolateral visual field and only correlates positively with the dorso ventral density of optical axes. On local stimulation, both the velocity tuning and the response latency of the LGMD/DCMD depend on stimulus position within the visual field. Spatial and temporal integration experiments in which several local motion stimuli were activated either simultaneously or at fixed delays reveal that the LGMD processes local motion in a strongly sublinear way. Thus the neuron's integration properties seem to depend on several factors including its dendritic morphology, the local characteristics of afferent fiber inputs, and inhibition mediated by different pathways or by voltage-gated conductances. Our study shows that the selectivity of this looming sensitive neuron to approaching objects relies on more complex biophysical mechanisms than previously thought. PMID- 15548623 TI - Integration of synaptic responses to neighboring whiskers in rat barrel cortex in vivo. AB - Characterizing input integration at the single-cell level is a critical step to understanding cortical function, particularly when sensory stimuli are represented over wide cortical areas and single cells exhibit large receptive fields. To study synaptic integration of sensory inputs, we made intracellular recordings from the barrel cortex of anesthetized rats in vivo. For each cell, we deflected the principal whisker (PW) either alone or preceded by the deflection of a single adjacent whisker (AW) at an interval of 20 or 3 ms. At the 20-ms interval in all cases, prior AW deflection significantly suppressed the PW-evoked spike output and caused the underlying synaptic response to reach a peak Vm less depolarized than that arising from PW deflection alone. The decrease in peak Vm was not attributed to hyperpolarizing inhibition but to a divisive reduction in PW-evoked PSP amplitude. The reduction in amplitude was not a result of shunting inhibition but was mostly a result of removal of the synaptic drive, or disfacilitation. When the AW-PW interval was shortened to 3 ms, spike suppression was observed in a subset of the cells studied. In most cases, a divisive reduction in synaptic response amplitude was offset by summation with the preceding AW-evoked depolarization. To determine whether suppression is a general feature of synaptic integration by barrel cortex neurons, we also characterized the interaction of responses evoked by local electrical stimulation. In contrast to the whisker data, we found that responses to paired stimulation at the same intervals produced more spikes and reached a peak Vm more depolarized than the individual responses alone, suggesting that whisker-evoked suppression is not a result of postsynaptic mechanisms. Instead, we propose that cross-whisker response suppression depends on sensory-specific mechanisms at cortical and subcortical levels. PMID- 15548624 TI - Acetylcholine dynamically controls spatial integration in marmoset primary visual cortex. AB - Recent in vitro studies have shown that acetylcholine (ACh) selectively reduces the efficacy of lateral cortical connections via a muscarinic mechanism, while boosting the efficacy of thalamocortical/feed-forward connections via a nicotinic mechanism. This suggests that high levels of ACh should reduce center-surround interactions of neurons in primary visual cortex, making cells more reliant on feed-forward information. In line with this hypothesis, we show that local iontophoretic application of ACh in primate primary visual cortex reduced the extent of spatial integration, assessed by recording a neurons' length tuning. When ACh was externally applied, neurons' preferred length shifted toward shorter bars, showing reduced impact of the extra-classical receptive field. We fitted a difference and a ratio of Gaussian model to these data to determine the underlying mechanisms of this dynamic change of spatial integration. These models assume overlapping summation and suppression areas with different widths and gains to be responsible for spatial integration and size tuning. ACh significantly reduced the extent of the summation area, but had no significant effect on the extent of the suppression area. In line with previous studies, we also show that applying ACh enhanced the response in the majority of cells, especially in the later (sustained) part of the response. These findings are similar to effects of attention on neuronal activity. The natural release of ACh is strongly linked with states of arousal and attention. Our results may therefore be relevant to the neurobiological mechanism of attention. PMID- 15548625 TI - Metabotropic glutamate receptor antagonist AIDA blocks induction of mossy fiber CA3 LTP in vivo. AB - Metabotropic glutamate receptors (mGluR) are implicated in long-term memory storage. mGluR-I and mGluR-II antagonists impede various forms of learning and long-term potentiation (LTP) in animals. Despite the evidence linking mGluR to learning mechanisms, their role in mossy fiber-CA3 long-term potentiation (LTP) is not yet clear. To explain the involvement of mGluR-I in memory mechanisms, we examined the function of the mGluR-I antagonist 1-aminoindan-1, 5-dicarboxylic acid (AIDA) on the induction of mossy fiber-CA3 LTP in vivo in male Sprague Dawley and Fischer 344 (F344) rats. Acute extracellular mossy fiber (MF) responses were evoked by stimulation of the MF bundle and recorded in the stratum lucidum of CA3. The excitatory postsynaptic potential (EPSP) magnitude was measured by using the initial slope of the field EPSP slope measured 2-3 ms after response onset. After collection of baseline MF-CA3 responses at 0.05 Hz, animals received either ((+/-))-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (N methyl-D-aspartate-R antagonist, 10 mg/kg ip), naloxone (opioid-R antagonist, 10 mg/kg ip), or AIDA (mGluR antagonist, 1 mg/kg ip or 37.5 nmol ic). LTP was induced by two 100-Hz trains at the intensity sufficient to evoke 50% of the maximal response. Responses were collected for an additional 1 h. AIDA blocked induction of LTP in the mossy fiber pathway (P < 0.05) in both strains of rats after systemic and in Sprague Dawley rats after intrahippocampal injection. PMID- 15548626 TI - Normal performance and expression of learning in the vestibulo-ocular reflex (VOR) at high frequencies. AB - The rotatory vestibulo-ocular reflex (VOR) keeps the visual world stable during head movements by causing eye velocity that is equal in amplitude and opposite in direction to angular head velocity. We have studied the performance of the VOR in darkness for sinusoidal angular head oscillation at frequencies ranging from 0.5 to 50 Hz. At frequencies of > or = 25 Hz, the harmonic distortion of the stimulus and response were estimated to be <14 and 22%, respectively. We measured the gain of the VOR (eye velocity divided by head velocity) and the phase shift between eye and head velocity before and after adaptation with altered vision. Before adaptation, VOR gains were close to unity for frequencies < or = 20 Hz and increased as a function of frequency reaching values of 3 or 4 at 50 Hz. Eye velocity was almost perfectly out of phase with head velocity for frequencies < or = 12.5 Hz, and lagged perfect compensation increasingly as a function of frequency. After adaptive modification of the VOR with magnifying or miniaturizing optics, gain showed maximal changes at frequencies <12.5 Hz, smaller changes at higher frequencies, and no change at frequencies larger than 25 Hz. Between 15 and 25 Hz, the phase of eye velocity led the unmodified VOR by as much as 50 degrees when the gain of the VOR had been decreased, and lagged when the gain of the VOR had been increased. We were able to reproduce the main features of our data with a two-pathway model of the VOR, where the two pathways had different relationships between phase shift and frequency. PMID- 15548627 TI - Sequences of predictive saccades are correlated over a span of approximately 2 s and produce a fractal time series. AB - We previously demonstrated that there is an abrupt (rather than smooth) transition between reactive and predictive modes of eye-movement tracking of target lights (a phase transition). We also found evidence that the sequence of eye movements in the reactive mode was independent, whereas those in the predictive mode were correlated and possibly formed a random fractal sequence. Here we confirm the finding of fractal structure by quantifying the rate of decay of nonlinear forecasting when applied to these data. We also estimate the window over which consecutive trials are correlated and show that the duration of this window is fixed in time rather than number of trials. These results have implications for the neural mechanisms that drive predictive movements. PMID- 15548628 TI - Using heterogeneity to predict inhibitory network model characteristics. AB - From modeling studies it has been known for >10 years that purely inhibitory networks can produce synchronous output given appropriate balances of intrinsic and synaptic parameters. Several experimental studies indicate that synchronous activity produced by inhibitory networks is critical to the production of population rhythms associated with various behavioral states. Heterogeneity of inputs to inhibitory networks strongly affect their ability to synchronize. In this paper, we explore how the amount of input heterogeneity to two-cell inhibitory networks affects their dynamics. Using numerical simulations and bifurcation analyses, we find that the ability of inhibitory networks to synchronize in the face of heterogeneity depends nonmonotonically on each of the synaptic time constant, synaptic conductance and external drive parameters. Because of this, an optimal set of parameters for a given cellular model with various biophysical characteristics can be determined. We suggest that this could be a helpful approach to use in determining the importance of different, underlying biophysical details. We further find that two-cell coherence properties are maintained in larger 10-cell networks. As such, we think that a strategy of "embedding" small network dynamics in larger networks is a useful way to understand the contribution of biophysically derived parameters to population dynamics in large networks. PMID- 15548629 TI - Using neuronal latency to determine sensory-motor processing pathways in reaction time tasks. AB - We describe a new technique that uses the timing of neuronal and behavioral responses to explore the contributions of individual neurons to specific behaviors. The approach uses both the mean neuronal latency and the trial-by trial covariance between neuronal latency and behavioral response. Reliable measurements of these values were obtained from single-unit recordings made from anterior inferotemporal (AIT) cortex and the frontal eye fields (FEF) in monkeys while they performed a choice reaction time task. These neurophysiological data show that the responses of AIT neurons and some FEF neurons have little covariance with behavioral response, consistent with a largely "sensory" response. The responses of another group of FEF neurons with longer mean latency covary tightly with behavioral response, consistent with a largely "motor" response. A very small fraction of FEF neurons had responses consistent with an intermediate position in the sensory-motor pathway. These results suggest that this technique is a valuable tool for exploring the functional organization of neuronal circuits that underlie specific behaviors. PMID- 15548630 TI - Modulatory effect of substance P to the brain stem locomotor command in lampreys. AB - Substance P initiates locomotion when injected in the brain stem of mammals. This study examined the possible role of this peptide on the supraspinal locomotor command system in lampreys. Substance P was bath applied or locally injected into an in vitro isolated brain stem, and the effects of the drug were examined on reticulospinal cells and on the occurrence of swimming in a semi-intact preparation. Bath applications of substance P induced sustained depolarizations occurring rhythmically in intracellularly recorded reticulospinal cells. Spiking activity was superimposed on the depolarizations and swimming was induced. The sustained depolarizations were abolished by tetrodotoxin, and substance P did not affect the membrane resistance of reticulospinal cells nor their firing properties, suggesting that it did not directly effect reticulospinal cells. To establish where the effects were exerted, successive lesions of the brain stem were made as well as local applications of the drug in the brain stem. Removing the mesencephalon abolished the sustained depolarizations, whereas large ejections of the drug in the mesencephalon excited reticulospinal cells and elicited bouts of swimming. More local injections into the mesencephalic locomotor region (MLR) also elicited swimming. After an injection of substance P, the current threshold needed to induce locomotion by MLR stimulation was decreased, and the size of the postsynaptic responses of reticulospinal cells to MLR stimulation was increased. Substance P also reduced the frequency of miniature spontaneous postsynaptic currents in reticulospinal cells. Taken together, these results suggest that substance P plays a neuromodulatory role on the brain stem locomotor networks of lampreys. PMID- 15548631 TI - Mechanisms determining the dynamic range of the bullfrog olfactory receptor cell. AB - Spike discharges of single olfactory receptor cells (ORCs) were recorded with the whole cell patch-clamp method applied to slice preparation. In parallel, activities of transduction channels were measured under the voltage-clamp condition. When cells were stimulated by odorants, 54 out of 306 cells exhibited inward current responses (10 mM cineole in the puffer pipette). The amplitude of the inward current was dependent on the stimulus period, reflecting the time integration for the stimulus dose, and the relation could be fitted by the Hill equation. Under the current-clamp condition, current injection induced spike discharges. In cells showing repetitive firings, the firing frequency was dependent on the amount of injected current. The relation was fitted by the Michaelis-Menten equation showing saturation. When cells were responsive to the odorant and had abilities to discharge repetitive spikes, the depolarizing responses were accompanied by repetitive spikes. In those cells, the spike frequency was dose-dependent, expressing saturation similar to the result obtained by current injection. Since both transduction channel and spike generative steps expressed saturation in their dose dependences, we explored what step(s) actually determines saturation in ORC signaling processes. By examining dose-response relations of both the current and spikes in the same cells, saturating dose was found to be dependent largely on that of the transduction step. This suggests that the dynamic range is fundamentally determined by the transduction system. In addition, a simple model derived from the nonlinearity of the plasma membrane could explain that a critical level of dynamic range was, at least in part, modified by the membrane nonlinearity. PMID- 15548632 TI - Closing in on the neural mechanisms of finger joint angle sense. Focus on "Quantitative analysis of dynamic strain sensitivity in human skin mechanoreceptors". PMID- 15548633 TI - Subtype-specific mechanisms for regulating K+ channel density during development. Focus on "The carboxyl tail region of the Kv2.2 subunit mediates novel developments of channel density". PMID- 15548634 TI - Lateral inhibition for center-surround reorganization of the frequency map of bat auditory cortex. AB - Repetitive acoustic stimulation, auditory fear conditioning, and focal electric stimulation of the auditory cortex (AC) each evoke the reorganization of the central auditory system. Our current study of the big brown bat indicates that focal electric stimulation of the AC evokes center-surround reorganization of the frequency map of the AC. In the center, the neuron's best frequencies (BFs), together with their frequency-tuning curves, shift toward the BFs of electrically stimulated cortical neurons (centripetal BF shifts). In the surround, BFs shift away from the stimulated cortical BF (centrifugal BF shifts). Centripetal BF shifts are much larger than centrifugal BF shifts. An antagonist (bicuculline methiodide) of inhibitory synaptic transmitter receptors changes centrifugal BF shifts into centripetal BF shifts, whereas its agonist (muscimol) changes centripetal BF shifts into centrifugal BF shifts. This reorganization of the AC thus depends on a balance between facilitation and inhibition evoked by focal cortical electric stimulation. Unlike neurons in the AC of the big brown bat, neurons in the Doppler-shifted constant-frequency (DSCF) area of the AC of the mustached bat are highly specialized for fine-frequency analysis and show almost exclusively centrifugal BF shifts for focal electric stimulation of the DSCF area. Our current data indicate that in the highly specialized area, lateral inhibition is strong compared with the less-specialized area and that the specialized and nonspecialized areas both share the same inhibitory mechanism for centrifugal BF shifts. PMID- 15548635 TI - Synaptic enhancement induced through callosal pathways in cat association cortex. AB - The corpus callosum plays a major role in synchronizing neocortical activities in the two hemispheres. We investigated the changes in callosally elicited excitatory postsynaptic potentials (EPSPs) of neurons from cortical association areas 5 and 7 of cats under barbiturate or ketamine-xylazine anesthesia. Single pulses to callosal pathway evoked control EPSPs; pulse-trains were subsequently applied at different frequencies to homotopic sites in the contralateral cortex, as conditioning stimulation; thereafter, the single pulses were applied again to test changes in synaptic responsiveness by comparing the amplitudes of control and conditioned EPSPs. In 41 of 42 neurons recorded under barbiturate anesthesia, all frequencies of conditioning callosal stimuli induced short-term (5-30 min) enhancement of test EPSPs elicited by single stimuli. Neurons tested with successive conditioning pulse-trains at different frequencies displayed stronger enhancement with high-frequency (40-100 Hz) than with low-frequency (10-20 Hz) rhythmic pulse-trains; >100 Hz, the potentiation saturated. In a neuronal sample, microdialysis of an N-methyl-D-aspartate (NMDA) receptor blocker in barbiturate treated cats suppressed this potentiation, and potentiation of callosally evoked EPSPs was not detected in neurons recorded under ketamine-xylazine anesthesia, thus indicating that EPSPs' potentiation implicates, at least partially, NMDA receptors. These data suggest that callosal activities occurring within low frequency and fast-frequency oscillations play a role in cortical synaptic plasticity. PMID- 15548636 TI - Quantitative analyses of dynamic strain sensitivity in human skin mechanoreceptors. AB - Microneurographical recordings from 24 slowly adapting (SA) and 16 fast adapting (FA) cutaneous mechanoreceptor afferents were obtained in the human radial nerve. Most of the afferents innervated the hairy skin on the back of the hand. The afferents' receptive fields were subjected to controlled strains in a ramp-and hold fashion with strain velocities from 1 to 64%.s(-1), i.e., strain velocities within most of the physiological range. For all unit types, the mean variation in response onset approached 1 ms for strain velocities >8%.s(-1). Except at the highest strain velocities, the first spike in a typical SAIII unit was evoked at strains <0.5% and a typical SAII unit began to discharge at <1% skin strain. Skin strain velocity had a profound effect on the discharge rates of all classes of afferents. The "typical" peak discharge rate at the highest strain velocity studied was 50-95 imp/s(-1) depending on unit type. Excellent fits were obtained for both SA and FA units when their responses to ramp stretches were modeled by simple power functions (r2 > 0.9 for 95% of the units). SAIII units grouped with SAII with respect to onset latency and onset variation but with SAI units with respect to dynamic strain sensitivity. Because both SA and FA skin afferents respond strongly, quickly, and accurately to skin strain changes, they all seem to be able to provide useful information about movement-related skin strain changes and therefore contribute to proprioception and kinesthesia. PMID- 15548638 TI - Cortico-motoneuronal system and dexterous finger movements. PMID- 15548637 TI - Afferent input modulates neurotrophins and synaptic plasticity in the spinal cord. AB - The effects of eliminating or decreasing neuromuscular activity on the expression of neurotrophins and associated molecules in the spinal cord and subsequent effects on spinal cord plasticity were determined. Spinal cord isolation (SI), which eliminates any supraspinal and peripheral monosynaptic input to the lumbar region but maintains the motoneuron-muscle connectivity, decreased the levels of brain-derived neurotrophic factor (BDNF) and neurotrophin 3 (NT-3) mRNA and protein in the isolated segments. Synapsin I, an important mediator for the effects of BDNF on synaptic plasticity, also was lower in the lumbar region of SI rats. In contrast, the levels of BDNF, synapsin, and growth-associated protein (GAP-43) were increased in the cervical spinal cord enlargement rostral to the isolated region, most likely reflecting an increased use of the forelimbs in the SI rats. GAP-43 levels were also increased in the lumbar spinal cord region, probably associated with compensatory mechanisms related to the deafferentation. In a separate set of experiments, the soleus muscle was paralyzed unilaterally via intramuscular botulinum toxin type A (BTX-A) injection to determine the effects of reducing the propioceptive input, of this normally highly active muscle on neurotrophin expression in the spinal cord. BDNF and synapsin I mRNAs were lower and NT-3 levels were higher in the lumbar hemicord ipsilateral to the BTX-A injection. Combined, these results indicate that the level of supraspinal and muscle afferent input plays an important role in modulating the levels of BDNF and NT-3 in the spinal cord. PMID- 15548639 TI - Deregulation of cell proliferation by polycyclic aromatic hydrocarbons in human breast carcinoma MCF-7 cells reflects both genotoxic and nongenotoxic events. AB - Polycyclic aromatic hydrocarbons (PAHs), such as benzo[a]pyrene (BaP), are carcinogens suggested to be involved in development of human cancer. Several recent studies have reported that PAHs can activate estrogen receptors (ER), either directly or indirectly by producing estrogenic metabolites. We hypothesized that the activation of ER by PAHs or their metabolites could induce cell proliferation in estrogen-sensitive cells. In the present study, we found that two PAHs, benz[a]anthracene (BaA) and BaP, can stimulate proliferation of human breast carcinoma MCF-7 cells at concentrations 100 nM and higher. This effect was ER-dependent, because it was blocked by the pure antiestrogen ICI 182,780. Although both PAHs partially inhibited S-phase entry and DNA synthesis induced by 17beta-estradiol, they stimulated S-phase entry when applied to MCF-7 cells synchronized by serum deprivation. This was in contrast with model antiestrogenic aryl hydrocarbon receptor ligand, 2,3,7,8-tetrachlorodibenzo-p dioxin, which fully suppressed S-phase entry. BaP, which is a strong mutagen, was found to induce p53 tumor suppressor expression, a partial S-phase arrest and at higher concentrations also cell death. Pifithrin-alpha, a synthetic inhibitor of p53 activity, abolished both S-phase arrest and apoptosis induced by genotoxic PAHs, and it potentiated the proliferative effect of BaP. Thus, both genotoxic and nongenotoxic events seem to interact in the effects of BaP on cell proliferation. Taken together, our data indicate that both BaA and BaP can stimulate cell proliferation through activation of ER. The proliferative effects of these carcinogenic compounds might contribute to tumor promotion in estrogen sensitive tissues. PMID- 15548640 TI - On the importance of exposure variability to the doses of volatile organic compounds. AB - The connection between occupational exposure to volatile organic compounds (VOCs) and the resulting internal doses is complicated by variability in air levels from day to day and by nonlinear kinetics of metabolism. We investigated long-term liver doses of VOCs and their metabolites using a physiologically based toxicokinetic model, to which 10,000 random 8-h exposures were inputted. Three carcinogenic VOCs were studied (i.e., benzene, perchloroethylene, and acrylonitrile); these compounds are all bioactivated in the liver and represent a wide range of an important toxicokinetic parameter Vmax/QL x KM. For each VOC, simulations were performed using mean air concentrations (muX) between 0.0003 and 1 mg/l (which covers both linear and saturated metabolism) and using coefficients of variation of exposure (CVX) between 0.23 and 2.18 (which includes most occupational settings). Two long-term measures of internal dose were examined, i.e., the area under the liver concentration-time curve (AUCL) and the area under the metabolic rate-time curve (AURC). Interestingly, both AUCL and AURC were linear functions of cumulative exposure (CE, mg x h/l air) even when metabolism was saturated and CVX was large. Yet, at a given CE, both AUCL and AURC were affected by CVX, with the magnitude of the effect increasing with Vmax/QL x KM (i.e., perchloroethylene < benzene < acrylonitrile). Nonetheless, the effects of CVX were typically only a few percent and should be of little consequence unless a VOC has large values of Vmax/QL x KM, muX,and CVX. We conclude that CE should be a sufficient predictor of the dose of either the parent chemical (VOC) or its metabolite in the liver, even when metabolism is nonlinear. We also observed that AUCL and AURC were sensitive to changes in values of model parameters in the high variability scenarios, suggesting that (when CVX is large) the population variability of AUCL and AURC can be quite large at a fixed CE. PMID- 15548641 TI - Acute respiratory responses of the mouse to chlorine. AB - In human subjects 15-min exposure to 0.5-1.0 ppm chlorine gas causes a nasal obstructive response in the absence of a marked sensation of irritation. The current investigation was designed to assess the response of the mouse for comparative purposes. Respiratory physiological responses were measured in female C57Bl/6J mice exposed to 0.8 to 4.0 ppm chlorine gas. Chlorine was a potent sensory irritant with an RD50 of 2.3 ppm. The gas produced airway obstruction as indicated by a concentration-dependent increase in specific airways resistance (sRaw) during the 15-min exposure. At 0.8 ppm, chlorine produced only mild sensory irritation (<20% change in breathing frequency) and a 65% increase in sRaw. Pretreatment with atropine was without effect on the obstructive response, suggesting a lack of involvement of muscarinic cholinergic pathways. Pretreatment with the sensory nerve toxin, capsaicin, dramatically reduced both the sensory irritation and obstructive responses to chlorine, suggesting the involvement of sensory nerves. Studies were also performed using the surgically isolated upper respiratory tract of the anesthetized mouse. Chlorine was efficiently scrubbed from the airstream (>97%) in that site and produced an obstructive response that was of sufficient magnitude to account for the entire response observed in the intact animal. In summary, chlorine gas produces an immediate nasal obstructive response in the mouse that appears to be similar to that in the human. PMID- 15548642 TI - D2 dopamine receptor-mediated modulation of voltage-dependent Na+ channels reduces autonomous activity in striatal cholinergic interneurons. AB - Striatal cholinergic interneurons are critical elements of the striatal circuitry controlling motor planning, movement, and associative learning. Intrastriatal release of dopamine and inhibition of interneuron activity is thought to be a critical link between behaviorally relevant events, such as reward, and alterations in striatal function. However, the mechanisms mediating this modulation are unclear. Using a combination of electrophysiological, molecular, and computational approaches, the studies reported here show that D2 dopamine receptor modulation of Na+ currents underlying autonomous spiking contributes to a slowing of discharge rate, such as that seen in vivo. Four lines of evidence support this conclusion. First, D2 receptor stimulation in tissue slices reduced the autonomous spiking in the presence of synaptic blockers. Second, in acutely isolated neurons, D2 receptor activation led to a reduction in Na+ currents underlying pacemaking. The modulation was mediated by a protein kinase C dependent enhancement of channel entry into a slow-inactivated state at depolarized potentials. Third, the sodium channel blocker TTX mimicked the effects of D2 receptor agonists on pacemaking. Fourth, simulation of cholinergic interneuron pacemaking revealed that a modest increase in the entry of Na+ channels into the slow-inactivated state was sufficient to account for the slowing of pacemaker discharge. These studies establish a cellular mechanism linking dopamine and the reduction in striatal cholinergic interneuron activity seen in the initial stages of associative learning. PMID- 15548643 TI - Structural abnormalities at neuromuscular synapses lacking multiple syntrophin isoforms. AB - The syntrophins are modular adapter proteins that function by recruiting signaling molecules to the cytoskeleton via their direct association with proteins of the dystrophin protein family. We investigated the physiological function of beta2-syntrophin by generating a line of mice lacking this syntrophin isoform. The beta2-syntrophin null mice show no overt phenotype, or muscular dystrophy, and form structurally normal neuromuscular junctions (NMJs). To determine whether physiological consequences caused by the lack of beta2 syntrophin were masked by compensation from the alpha-syntrophin isoform, we crossed these mice with our previously described alpha-syntrophin null mice to produce mice lacking both isoforms. The alpha/beta2-syntrophin null mice have NMJs that are structurally more aberrant than those lacking only alpha syntrophin. The NMJs of the alpha/beta2-syntrophin null mice have fewer junctional folds than either parent strain, and the remaining folds are abnormally shaped with few openings to the synaptic space. The levels of acetylcholine receptors are reduced to 23% of wild type in mice lacking both syntrophin isoforms. Furthermore, the alpha/beta2-syntrophin null mice ran significantly shorter distances on voluntary exercise wheels despite having normal neuromuscular junction transmission as determined by micro-electrode recording of endplate potentials. We conclude that both alpha-syntrophin and beta2-syntrophin play distinct roles in forming and maintaining NMJ structure and that each syntrophin can partially compensate for the loss of the other. PMID- 15548644 TI - The subcellular organization of cortactin in hippocampus. AB - Spines may undergo rapid, activity-dependent changes in shape and size, reflecting reorganization of the actin cytoskeleton. This remodeling is implicated in development and also in the late phase of long-term potentiation. However, the cellular mechanisms that convert activity into morphological change remain poorly understood, and little is known about the anatomical distribution of the actin-regulating proteins that mediate this remodeling. Using immunocytochemistry, we demonstrate here that cortactin (a protein implicated in actin filament nucleation, branching, and stabilization) is concentrated in hippocampal spines, where it colocalizes with F-actin. Cortactin has a Shank binding domain; recent studies report that synaptic activity may trigger actin remodeling via this interaction with Shank. However, our immunogold electron microscopic data show that cortactin concentrates within the spine core, 100-150 nm away from the postsynaptic density (PSD); only a small fraction of the cortactin in spines lies adjacent to the PSD. These data suggest that the adult dendritic spine contains two functional pools of cortactin: a large pool in the spine core that may help to mediates changes in spine shape and a small synaptic pool that may modify the PSD in response to synaptic activity. PMID- 15548645 TI - Molecular mechanism of pregnenolone sulfate action at NR1/NR2B receptors. AB - NMDA receptors are highly expressed in the CNS and are involved in excitatory synaptic transmission and synaptic plasticity as well as excitotoxicity. They have several binding sites for allosteric modulators, including neurosteroids, endogenous compounds synthesized by the nervous tissue and expected to act locally. Whole-cell patch-clamp recording from human embryonic kidney 293 cells expressing NR1-1a/NR2B receptors revealed that neurosteroid pregnenolone sulfate (PS) (300 microm), when applied to resting NMDA receptors, potentiates the amplitude of subsequent responses to 1 mm glutamate fivefold and slows their deactivation twofold. The same concentration of PS, when applied during NMDA receptor activation by 1 mm glutamate, has only a small effect. The association and dissociation rate constants of PS binding and unbinding from resting NMDA receptors are estimated to be 3.3 +/- 2.0 mm(-1)sec(-1) and 0.12 +/- 0.02 sec( 1), respectively, corresponding to an apparent affinity K(d) of 37 microm. The results of experiments indicate that the molecular mechanism of PS potentiation of NMDA receptor responses is attributable to an increase in the peak channel open probability (P(o)). Responses to glutamate recorded in the continuous presence of PS exhibit marked time-dependent decline. Our results indicate that the decline is induced by a change of the NMDA receptor affinity for PS after receptor activation. These results suggest that the PS is a modulator of NMDA receptor P(o), the effectiveness of which is lowered by glutamate binding. This modulation may have important consequences for the neuronal excitability. PMID- 15548646 TI - Fear-potentiated startle in rats is mediated by neurons in the deep layers of the superior colliculus/deep mesencephalic nucleus of the rostral midbrain through the glutamate non-NMDA receptors. AB - The amygdala sends heavy and broad projections to the rostral midbrain including the periaqueductal gray (PAG), the deep layers of the superior colliculus/deep mesencephalic nucleus (deep SC/DpMe), and the lateral mesencephalic reticular formation (MRF) that in turn project to the nucleus reticularis pontis caudalis (PnC), an obligatory relay in the primary acoustic startle circuit. Chemical lesions or inactivation of these areas blocked fear-potentiated startle, suggesting that these areas serve as a relay between the amygdala and the PnC. In the present study, we tried to determine more precisely which of these sites were critical for fear-potentiated startle and the role of glutamate receptors in this site in mediating fear-potentiated startle. Local infusion of the non-NMDA receptor antagonist 2,3-dihydroxy-6-nitro-7-sulfamoylbenzo(F)-quinoxaline (NBQX) dose-dependently blocked fear-potentiated startle when infused into the deep SC/DpMe before testing but had no effect on baseline startle amplitude. NBQX did not block fear-potentiated startle when infused before training. The same dose of NBQX infused into the dorsal/lateral PAG, the lateral MRF, or the superficial layers of the SC did not affect fear-potentiated startle. However, NBQX tended to reduce contextual freezing when infused into the dorsal/lateral PAG. These findings suggest that the deep SC/DpMe is the site that serves as a critical output relay between the amygdala and the PnC in mediating fear-potentiated startle and that glutamatergic transmission is required for this action. PMID- 15548648 TI - PHCCC, a specific enhancer of type 4 metabotropic glutamate receptors, reduces proliferation and promotes differentiation of cerebellar granule cell neuroprecursors. AB - Exposure of immature rat cerebellar granule cell cultures to the type 4 metabotropic glutamate (mGlu4) receptor enhancer N-phenyl-7 (hydroxyimino)cyclopropa[b]chromen-1a-carboxamide (PHCCC) reduced [3H]thymidine incorporation. Its action was sensitive to the growth conditions and was attenuated by two mGlu4 receptor antagonists. An antiproliferative action of PHCCC was also seen in cultures from wild-type, but not mGlu4, knock-out mice. At least in rat cultures, PHCCC was not neurotoxic and enhanced neuritogenesis. Although PHCCC reduced the increase in cAMP formation and phospho-AKT levels induced by forskolin, none of these transduction pathways significantly contributed to the reduction of [3H]thymidine incorporation. Interestingly, PHCCC reduced the expression of Gli-1, a transcription factor that mediates the mitogenic action of Sonic hedgehog. Finally, we treated newborn rats with PHCCC either intracerebrally (infusion of 5 nmol/2 microl in the cerebellar region once every other day) or systemically (5 mg/kg, i.p., once daily) from postnatal days 3-9. Local infusion of PHCCC induced substantial changes in the morphology of the developing cerebellum. In contrast, systemic injection of PHCCC induced only morphological abnormalities of the cerebellar lobule V, which became visible 11 d after the end of the treatment. These data suggest that mGlu4 receptors are involved in the regulation of cerebellar development. PMID- 15548647 TI - Chemotherapy for the brain: the antitumor antibiotic mithramycin prolongs survival in a mouse model of Huntington's disease. AB - Huntington's disease (HD) is a fully penetrant autosomal-dominant inherited neurological disorder caused by expanded CAG repeats in the Huntingtin gene. Transcriptional dysfunction, excitotoxicity, and oxidative stress have all been proposed to play important roles in the pathogenesis of HD. This study was designed to explore the therapeutic potential of mithramycin, a clinically approved guanosine-cytosine-rich DNA binding antitumor antibiotic. Pharmacological treatment of a transgenic mouse model of HD (R6/2) with mithramycin extended survival by 29.1%, greater than any single agent reported to date. Increased survival was accompanied by improved motor performance and markedly delayed neuropathological sequelae. To identify the functional mechanism for the salubrious effects of mithramycin, we examined transcriptional dysfunction in R6/2 mice. Consistent with transcriptional repression playing a role in the pathogenesis of HD, we found increased methylation of lysine 9 in histone H3, a well established mechanism of gene silencing. Mithramycin treatment prevented the increase in H3 methylation observed in R6/2 mice, suggesting that the enhanced survival and neuroprotection might be attributable to the alleviation of repressed gene expression vital to neuronal function and survival. Because it is Food and Drug Administration-approved, mithramycin is a promising drug for the treatment of HD. PMID- 15548650 TI - State anxiety modulation of the amygdala response to unattended threat-related stimuli. AB - Findings from fear-conditioning studies in rats and functional neuroimaging with human volunteers have led to the suggestion that the amygdala is involved in the preattentive detection of threat-related stimuli. However, some neuroimaging findings point to attentional modulation of the amygdala response. The clinical cognitive literature suggests that the extent to which the processing of threat related stimuli is modulated by attention is crucially dependent on participants' anxiety levels. Here, we conducted a functional magnetic resonance imaging study with 27 healthy volunteers to examine whether amygdala responsivity to unattended threat-related stimuli varies with individual differences in state anxiety. Pairs of houses and faces (both fearful or neutral in expression) were presented, and participants attended to either the faces or the houses and matched these stimuli on identity. "Low-anxious" participants showed a reduced amygdala response to unattended versus attended fearful faces, but "high-anxious" participants showed no such reduction, having an increased amygdala response to fearful versus neutral faces regardless of attentional focus. These findings suggest that anxiety may interact with attentional focus to determine the magnitude of the amygdala response to threat-related stimuli. PMID- 15548649 TI - Vascular endothelial cells promote acute plasticity in ependymoglial cells of the neuroendocrine brain. AB - Glial and endothelial cells interact throughout the brain to define specific functional domains. Whether endothelial cells convey signals to glia in the mature brain is unknown but is amenable to examination in circumventricular organs. Here we report that purified endothelial cells of one of these organs, the median eminence of the hypothalamus, induce acute actin cytoskeleton remodeling in isolated ependymoglial cells and show that this plasticity is mediated by nitric oxide (NO), a diffusible factor. We found that both soluble guanylyl cyclase and cyclooxygenase products are involved in this endothelial mediated control of ependymoglia cytoarchitecture. We also demonstrate by electron microscopy that activation of endogenous NO release in the median eminence induces rapid structural changes, allowing a direct access of neurosecretory axons containing gonadotropin-releasing hormone (GnRH) (the neuropeptide controlling reproductive function) to the portal vasculature. Local in vivo inhibition of NO synthesis disrupts reproductive cyclicity, a process that requires a pulsatile, coordinated delivery of GnRH into the hypothalamic adenohypophyseal portal system. Our results identify a previously unknown function for endothelial cells in inducing neuroglial plasticity and raise the intriguing possibility that endothelial cells throughout the brain may use a similar signaling mechanism to regulate glial-neuronal interactions. PMID- 15548651 TI - Short- and long-term effects of cholinergic modulation on gamma oscillations and response synchronization in the visual cortex. AB - Neurons can engage in synchronized oscillatory activity in the gamma-frequency range when responding to sensory stimuli. Both the oscillatory patterning and the synchronization of responses are enhanced with arousal and attention or when the electroencephalogram is activated by electrical stimulation of the mesencephalic reticular formation. Here we show with intracortical application of cholinergic antagonists that the enhancement of gamma oscillations and response synchronization is mediated by acetylcholine and muscarinic receptors. We demonstrate further that coapplication of cholinergic agonists with synchrony inducing light stimuli causes a lasting increase in the probability that the stimulated cells engage in gamma oscillations and response synchronization. These changes develop slowly over tens of minutes and then persist for many hours. Thus, cholinergic modulation plays a crucial role both in the fast, state dependent facilitation of gamma oscillations and response synchronization and in use-dependent long-term modifications of cortical dynamics that favor synchronization of responses in the gamma-frequency range. PMID- 15548652 TI - Functional compensation of P/Q by N-type channels blocks short-term plasticity at the calyx of Held presynaptic terminal. AB - Calcium channels of the P/Q subtype mediate transmitter release at the neuromuscular junction and at many central synapses, such as the calyx of Held. Transgenic mice in which alpha1A channels are ablated provide a powerful tool with which to test compensatory mechanisms at the synapse and to explore mechanisms of presynaptic regulation associated with expression of P/Q channels. Using the calyx of Held preparation from the knock-out (KO) mice, we show here that N-type channels functionally compensate for the absence of P/Q subunits at the calyx and evoke giant synaptic currents [approximately two-thirds of the magnitude of wild-type (WT) responses]. However, although evoked paired-pulse facilitation is prominent in WT, this facilitation is greatly diminished in the KO. In addition, direct recording of presynaptic calcium currents revealed that the major functional difference was the absence of calcium-dependent facilitation at the calyx in the P/Q KO animals. We conclude that one physiological function of P/Q channels is to provide additional facilitatory drive, so contributing to maintenance of transmission as vesicles are depleted during high throughput synaptic transmission. PMID- 15548653 TI - Anosmin-1 modulates fibroblast growth factor receptor 1 signaling in human gonadotropin-releasing hormone olfactory neuroblasts through a heparan sulfate dependent mechanism. AB - Defects of either anosmin-1 or fibroblast growth factor receptor 1 (FGFR1) are known to underlie hereditary Kallmann's syndrome (KS), a human disorder of olfactory and gonadotropin-releasing hormone (GnRH) neuronal ontogeny. Here, we report a functional interaction between anosmin-1 and the FGFR1-FGF2-heparan sulfate complex, leading to amplified responses in the FGFR1 signaling pathway. In human embryonic GnRH olfactory neuroblasts, wild-type anosmin-1, but not proteins with loss-of-function KS mutations, induces neurite outgrowth and cytoskeletal rearrangements through FGFR1-dependent mechanisms involving p42/44 and p38 mitogen-activated protein kinases and Cdc42/Rac1 activation. Furthermore, anosmin-1 enhances FGF2 signaling specifically through FGFR1 IIIc in heterologous BaF3 lymphoid cells in a heparan sulfate-dependent manner. Our study provides compelling evidence for anosmin-1 as an isoform-specific co-ligand modulator of FGFR signaling that amplifies and specifies FGFR1 signaling responses during human nervous system development and defines a mechanism underlying the link between autosomal and X-linked KS. PMID- 15548654 TI - Memory-like alterations in Aplysia axons after nerve injury or localized depolarization. AB - Adaptive, long-term alterations of excitability have been reported in dendrites and presynaptic terminals but not along axons. Persistent enhancement of axonal excitability has been described in proximal nerve stumps at sites of nerve section in mammals, but this hyperexcitability is considered a pathological derangement important only as a cause of neuropathic pain. Identified neurons in Aplysia were used to test the hypothesis that either axonal injury or the focal depolarization that accompanies axonal injury can trigger a local decrease in action potential threshold [long-term hyperexcitability (LTH)] having memory-like properties. Nociceptive tail sensory neurons and a giant secretomotor neuron, R2, exhibited localized axonal LTH lasting 24 hr after a crush of the nerve or connective that severed the tested axons. Axons of tail sensory neurons and tail motor neurons, but not R2, displayed similar localized LTH after peripheral depolarization produced by 2 min exposure to elevated extracellular [K(+)]. Neither the induction nor expression of either form of LTH was blocked by saline containing 1% normal [Ca(2+)] during treatment or testing. However, both were prevented by local application of the protein synthesis inhibitors anisomycin or rapamycin. The features of (1) long-lasting alteration by localized depolarization, (2) restriction of alterations to intensely depolarized regions, and (3) dependence of the alterations on local, rapamycin-sensitive protein synthesis are shared with synaptic mechanisms considered important for memory formation. This commonality suggests that relatively simple, accessible axons may offer an opportunity to define fundamental plasticity mechanisms that were important in the evolution of memory. PMID- 15548655 TI - Presynaptic Cav2.1 and Cav2.2 differentially influence release dynamics at hippocampal excitatory synapses. AB - Presynaptic calcium influx at most excitatory central synapses is carried by both Cav2.1 and Cav2.2 channels. The kinetics and modulation of Cav2.1 and Cav2.2 channels differ and may affect presynaptic calcium influx. We compared release dynamics at CA3/CA1 synapses in rat hippocampus after selective blockade of either channel subtype and subsequent quantal content restoration. Selective blockade of Cav2.1 channels enhanced paired-pulse facilitation, whereas blockade of Cav2.2 channels decreased it. This effect was observed at short (50 msec) but not longer (500 msec) intervals and was maintained during prolonged bursts of presynaptic activity. It did not reflect differences in the distance of the channels from the calcium sensor. The suppression of this effect by preincubation with the G(o/i)-protein inhibitor pertussis toxin suggests instead that high frequency stimulation relieves inhibition of Cav2.2 by G(o/i), thereby increasing the number of available channels. PMID- 15548656 TI - Chronic back pain is associated with decreased prefrontal and thalamic gray matter density. AB - The role of the brain in chronic pain conditions remains speculative. We compared brain morphology of 26 chronic back pain (CBP) patients to matched control subjects, using magnetic resonance imaging brain scan data and automated analysis techniques. CBP patients were divided into neuropathic, exhibiting pain because of sciatic nerve damage, and non-neuropathic groups. Pain-related characteristics were correlated to morphometric measures. Neocortical gray matter volume was compared after skull normalization. Patients with CBP showed 5-11% less neocortical gray matter volume than control subjects. The magnitude of this decrease is equivalent to the gray matter volume lost in 10-20 years of normal aging. The decreased volume was related to pain duration, indicating a 1.3 cm3 loss of gray matter for every year of chronic pain. Regional gray matter density in 17 CBP patients was compared with matched controls using voxel-based morphometry and nonparametric statistics. Gray matter density was reduced in bilateral dorsolateral prefrontal cortex and right thalamus and was strongly related to pain characteristics in a pattern distinct for neuropathic and non neuropathic CBP. Our results imply that CBP is accompanied by brain atrophy and suggest that the pathophysiology of chronic pain includes thalamocortical processes. PMID- 15548657 TI - Stimulus and potassium-induced epileptiform activity in the human dentate gyrus from patients with and without hippocampal sclerosis. AB - Hippocampal specimens resected to cure medically intractable temporal lobe epilepsy (TLE) provide a unique possibility to study functional consequences of morphological alterations. One intriguing alteration predominantly observed in cases of hippocampal sclerosis is an uncommon network of granule cells monosynaptically interconnected via aberrant supragranular mossy fibers. We investigated whether granule cell populations in slices from sclerotic and nonsclerotic hippocampi would develop ictaform activity when challenged by low frequency hilar stimulation in the presence of elevated extracellular potassium concentration (10 and 12 mm) and whether the experimental activity differs according to the presence of aberrant mossy fibers. We found that ictaform activity could be evoked in slices from sclerotic and nonsclerotic hippocampi (27 of 40 slices, 14 of 20 patients; and 11 of 22 slices, 6 of 12 patients, respectively). However, the two patient groups differed with respect to the pattern of ictaform discharges and the potassium concentration mandatory for its induction. Seizure-like events were already induced with 10 mm K+. They exclusively occurred in slices from sclerotic hippocampi, of which 80% displayed stimulus-induced oscillatory population responses (250-300 Hz). In slices from nonsclerotic hippocampi, atypical negative field potential shifts were predominantly evoked with 12 mm K+. In both groups, the ictaform activity was sensitive to ionotropic glutamate receptor antagonists and lowering of [Ca2+]o. Our results show that, in granule cell populations of hippocampal slices from TLE patients, high K+-induced seizure-like activity and ictal spiking coincide with basic electrophysiological abnormalities, hippocampal sclerosis, and mossy fiber sprouting, suggesting that network reorganization could play a crucial role in determining type and threshold of such activity. PMID- 15548658 TI - "What" becoming "where": functional magnetic resonance imaging evidence for pragmatic relevance driving premotor cortex. AB - Previous studies using the serial prediction task (SPT) have shown that attending to the locations of objects activates the dorsal part of premotor cortex more than attending to the sizes of objects. The opposite holds for the ventral part of the premotor cortex. The present study used functional magnetic resonance imaging to investigate whether the learning of arbitrary stimulus-response mappings influences this functional dissociation. One experimental group learned to assign stimuli to response buttons based on stimulus size; another group did so based on stimulus location. More specifically, one-half of the participants in both experimental groups learned to assign stimuli to finger movements of their right hand, whereas the other half assigned stimuli to finger movements of their left hand. During scanning, all participants performed both size SPT and location SPT. Thus, we investigated the effects of the attended stimulus property (size or location), the motor effector assigned to it (fingers of left or right hand), and the spatial arrangement of the targets (the same in all groups). As expected, without motor training, the dorsal premotor cortex was less activated during size SPT compared with location SPT. The opposite held for ventral premotor cortex. With motor training, however, this differential activity pattern vanished. Activity in dorsal premotor cortex reflected neither the attended stimulus property nor the motor effector assigned to it. Instead, its activity may be related to the spatial properties of the response targets once some object property, such as size, takes on the "pragmatic relevance" of a spatially directed response. PMID- 15548659 TI - Multiple time scales of adaptation in auditory cortex neurons. AB - Neurons in primary auditory cortex (A1) of cats show strong stimulus-specific adaptation (SSA). In probabilistic settings, in which one stimulus is common and another is rare, responses to common sounds adapt more strongly than responses to rare sounds. This SSA could be a correlate of auditory sensory memory at the level of single A1 neurons. Here we studied adaptation in A1 neurons, using three different probabilistic designs. We showed that SSA has several time scales concurrently, spanning many orders of magnitude, from hundreds of milliseconds to tens of seconds. Similar time scales are known for the auditory memory span of humans, as measured both psychophysically and using evoked potentials. A simple model, with linear dependence on both short-term and long-term stimulus history, provided a good fit to A1 responses. Auditory thalamus neurons did not show SSA, and their responses were poorly fitted by the same model. In addition, SSA increased the proportion of failures in the responses of A1 neurons to the adapting stimulus. Finally, SSA caused a bias in the neuronal responses to unbiased stimuli, enhancing the responses to eccentric stimuli. Therefore, we propose that a major function of SSA in A1 neurons is to encode auditory sensory memory on multiple time scales. This SSA might play a role in stream segregation and in binding of auditory objects over many time scales, a property that is crucial for processing of natural auditory scenes in cats and of speech and music in humans. PMID- 15548660 TI - Nitric oxide synthase (NOS)-interacting protein interacts with neuronal NOS and regulates its distribution and activity. AB - Mechanisms governing the activity of neuronal nitric oxide synthase (nNOS), the major source of nitric oxide (NO) in the nervous system, are not completely understood. We report here a protein-protein interaction between nNOS and NOSIP (nitric oxide synthase-interacting protein) in rat brain in vivo. NOSIP and nNOS are concentrated in neuronal synapses and demonstrate significant colocalization in various regions of the central and peripheral nervous systems. NOSIP produces a significant reduction in nNOS activity in a neuroepithelioma cell line stably expressing nNOS. Furthermore, overexpression of NOSIP in cultured primary neurons reduces the availability of nNOS in terminal dendrites. These results thus suggest that the interaction between NOSIP and nNOS is functionally involved in endogenous mechanisms regulating NO synthesis. Furthermore, we found that the subcellular distribution and expression levels of NOSIP are dynamically regulated by neuronal activity in vitro as well as in vivo, suggesting that NOSIP may contribute to a mechanism via which neuronal activity regulates the synaptic availability and activity of nNOS. PMID- 15548661 TI - Increased expression of the Drosophila vesicular glutamate transporter leads to excess glutamate release and a compensatory decrease in quantal content. AB - Quantal size is a fundamental parameter controlling the strength of synaptic transmission. The transmitter content of synaptic vesicles is one mechanism that can affect the physiological response to the release of a single vesicle. At glutamatergic synapses, vesicular glutamate transporters (VGLUTs) are responsible for filling synaptic vesicles with glutamate. To investigate how VGLUT expression can regulate synaptic strength in vivo, we have identified the Drosophila vesicular glutamate transporter, which we name DVGLUT. DVGLUT mRNA is expressed in glutamatergic motoneurons and a large number of interneurons in the Drosophila CNS. DVGLUT protein resides on synaptic vesicles and localizes to the presynaptic terminals of all known glutamatergic neuromuscular junctions as well as to synapses throughout the CNS neuropil. Increasing the expression of DVGLUT in motoneurons leads to an increase in quantal size that is accompanied by an increase in synaptic vesicle volume. At synapses confronted with increased glutamate release from each vesicle, there is a compensatory decrease in the number of synaptic vesicles released that maintains normal levels of synaptic excitation. These results demonstrate that (1) expression of DVGLUT determines the size and glutamate content of synaptic vesicles and (2) homeostatic mechanisms exist to attenuate the excitatory effects of excess glutamate release. PMID- 15548662 TI - Time-resolved measurement of state-specific P2X2 ion channel cytosolic gating motions. AB - ATP-gated P2X2 channels undergo permeability changes through a process that is incompletely understood. In the present study, we used fluorescence resonance energy transfer (FRET) and electrophysiology to measure cytosolic gating motions in P2X2 channels as they enter a state with increased permeability. P2X2 channels underwent permeability changes with a time course that was similar to decreases in FRET between cyan fluorescent protein and yellow fluorescent protein attached to the cytosolic domain of P2X2 channels. Wild-type and mutant channels that did not undergo permeability changes also did not show evidence of cytosolic gating motions. Moreover, immobilizing the cytosolic domain by tethering it to the plasma membrane prevented the switch in permeability and impaired the cytosolic gating motions. Both of these phenotypes were restored when the immobilizing tether was cleaved. The data provide a time-resolved measure of state-specific gating motions and suggest how a cytosolic domain may control ion channel permeability. PMID- 15548663 TI - Memory consolidation and reconsolidation in the rat pup require protein synthesis. AB - Little is known about the ontogenesis of memory, whether it appears with its full characteristics or whether they emerge progressively with development. In the adult, basic characteristics of memory processing are consolidation of memory after acquisition and reconsolidation after retrieval. Here, using a conditioned aversion paradigm and postlearning or postreactivation injection of a protein synthesis inhibitor, we show that memory acquired by rat pups as early as postnatal day 3 requires time-dependent protein synthesis after both learning and reactivation. These results present the first evidence suggesting that consolidation and reconsolidation are original properties of memory function. PMID- 15548664 TI - Orexin neurons function in an efferent pathway of a food-entrainable circadian oscillator in eliciting food-anticipatory activity and wakefulness. AB - Temporal restriction of feeding can entrain circadian behavioral and physiological rhythms in mammals. Considering the critical functions of the hypothalamic orexin (hypocretin) neuropeptides in promoting wakefulness and locomotor activity, we examined the role of orexin neurons in the adaptation to restricted feeding. In orexin neuron-ablated transgenic mice, the food-entrained rhythmicity of mPer2 expression in the brain and liver, the reversal of the sleep wake cycle, and the recovery of daily food intake were unaltered compared with wild-type littermates. In contrast, orexin neuron-ablated mice had a severe deficit in displaying the normal food-anticipatory increases in wakefulness and locomotor activity under restricted feeding. Moreover, activity of orexin neurons markedly increased during the food-anticipatory period under restricted feeding in wild-type mice. Orexin neurons thus convey an efferent signal from putative food-entrainable oscillator or oscillators to increase wakefulness and locomotor activity. PMID- 15548665 TI - The role of palmitoylation in functional expression of nicotinic alpha7 receptors. AB - Neuronal alpha-bungarotoxin receptors (BgtRs) are nicotinic receptors that require as yet unidentified post-translational modifications to achieve functional expression. In this study, we examined the role of protein palmitoylation in BgtR expression. BgtR alpha7 subunits are highly palmitoylated in neurons from brain and other cells capable of BgtR expression, such as pheochromocytoma 12 (PC12) cells. In PC12 cells, alpha7 subunits are palmitoylated with a stoichiometry of approximately one palmitate per subunit, and inhibition of palmitoylation blocks BgtR expression. In cells incapable of BgtR expression, such as human embryonic kidney cells, alpha7 subunits are not significantly palmitoylated. However, in these same cells, chimeric subunits with the N-terminal half of alpha7 fused to the C-terminal half of serotonin-3A receptor (alpha7/5-HT3A) subunits form functional BgtRs that are palmitoylated to an extent similar to that of BgtRalpha7 subunits in PC12 cells. Palmitoylation of PC12 and alpha7/5-HT3A BgtRs occurred during assembly in the endoplasmic reticulum (ER). In conclusion, our data indicate a function for protein palmitoylation in which palmitoylation of assembling alpha7 subunits in the ER has a role in the formation of functional BgtRs. PMID- 15548666 TI - Blockade of Nogo-66, myelin-associated glycoprotein, and oligodendrocyte myelin glycoprotein by soluble Nogo-66 receptor promotes axonal sprouting and recovery after spinal injury. AB - The growth of injured axons in the adult mammalian CNS is limited after injury. Three myelin proteins, Nogo, MAG (myelin-associated glycoprotein), and OMgp (oligodendrocyte myelin glycoprotein), bind to the Nogo-66 receptor (NgR) and inhibit axonal growth in vitro. Transgenic or viral blockade of NgR function allows axonal sprouting in vivo. Here, we administered the soluble function blocking NgR ectodomain [aa 27-310; NgR(310)ecto] to spinal-injured rats. Purified NgR(310)ecto-Fc protein was delivered intrathecally after midthoracic dorsal over-hemisection. Axonal sprouting of corticospinal and raphespinal fibers in NgR(310)ecto-Fc-treated animals correlates with improved spinal cord electrical conduction and improved locomotion. The ability of soluble NgR(310)ecto to promote axon growth and locomotor recovery demonstrates a therapeutic potential for NgR antagonism in traumatic spinal cord injury. PMID- 15548667 TI - Neurotrophin signaling through the p75 receptor is deficient in traf6-/- mice. AB - Activation of the neurotrophin receptor p75 has been shown to elicit opposing cellular signals. Depending on the context of the cell, p75 will either promote survival or induce apoptosis after neurotrophin stimulation. p75-induced apoptosis occurs through activation of c-Jun N-terminal kinase (JNK), whereas the survival signal is mediated by nuclear factor kappaB (NFkappaB). The receptor proximal signals that produce these responses are unknown, although several molecules have been identified that associate with the intracellular domain of p75. One such interactor, TRAF6, a member of the tumor necrosis factor receptor associated factor family, has been implicated in p75 signaling. To assess the role of TRAF6 in p75 signaling, we analyzed mice with this gene deleted. In Schwann cells isolated from traf6+/+ animals, NGF elicited an 80% increase in transcription of an NFkappaB reporter; however, in traf6-/- cells, the NGF response was abrogated. Similarly, NGF activation of JNK was not apparent in Schwann cells from mice lacking traf6. Deficiencies in p75 signaling in traf6-/- animals resulted in a loss of p75-mediated apoptosis. In sympathetic neurons cultured from traf6+/+ superior cervical ganglia (SCGs), there was an increase in JNK activation and apoptosis after BDNF binding to p75; however, traf6-/- neurons did not respond. In vivo during naturally occurring cell death, there was a 55.6% reduction in TUNEL (terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling)-positive cells in the SCG of postnatal day 4 traf6-/- animals relative to traf6+/+ littermates. These results indicate that TRAF6 plays an essential role in mediating p75 signal transduction and induction of apoptosis. PMID- 15548668 TI - Postnatal neurogenesis and gliogenesis in the olfactory bulb from NG2-expressing progenitors of the subventricular zone. AB - We used a 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP)-enhanced green fluorescent protein (EGFP) transgenic mouse to study postnatal subventricular zone (SVZ) progenitor fate, with a focus on the olfactory bulb (OB). The postnatal OB of the CNP-EGFP mouse contained EGFP+ interneurons and oligodendrocytes. In the anterior SVZ, the majority of EGFP+ progenitors were NG2+. These NG2+/EGFP+ progenitors expressed the OB interneuron marker Er81, the neuroblast markers doublecortin (DC) and Distalless-related homeobox (DLX), or the oligodendrocyte progenitor marker Nkx2.2. In the rostral migratory stream (RMS), EGFP+ cells displayed a migrating phenotype. A fraction of these cells were either NG2-/Er81+/DC+/DLX+ or NG2+/Nkx2.2+. DiI (1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate) injection into the lateral ventricle (LV) of early postnatal mice demonstrated that NG2+/EGFP+ progenitors migrate from the SVZ through the RMS into the OB. Moreover, fluorescence-activated cell sorting-purified NG2+/CNP-EGFP+ or NG2+/beta-actin-enhanced yellow fluorescent protein-positive (EYFP+) progenitors transplanted into the early postnatal LV displayed extensive rostral and caudal migration. EYFP+ or EGFP+ graft-derived cells within the RMS were DLX+/Er81+ or Nkx2.2+, migrated to the OB, and differentiated to interneurons and oligodendrocytes. In the subcortical white matter (SCWM), grafted cells differentiated to either oligodendrocytes or astrocytes. Transplantation of NG2+/EYFP+ progenitors selectively purified from the SVZ showed that these cells were migratory and generated glia and neurons in the OB, hippocampus, and striatum. In contrast, cortical, OB, or cerebellar NG2+ cells had a very limited migratory potential and gave rise to glia in the SCWM and striatum. Our findings indicate region-specific differences between NG2+ progenitor cells and show that NG2+ cells can migrate throughout the RMS and contribute to both gliogenesis and neurogenesis in the postnatal OB. PMID- 15548669 TI - RACK1 and brain-derived neurotrophic factor: a homeostatic pathway that regulates alcohol addiction. AB - Alcoholism is a devastating disease that manifests as uncontrolled drinking. Consumption of alcohol is regulated by neurochemical systems within specific neural circuits, but endogenous systems that may counteract and thus suppress the behavioral effects of ethanol intake are unknown. Here we demonstrate that BDNF plays a role in reducing the behavioral effects of ethanol, including consumption, in rodents. We found that decreasing the levels of BDNF leads to increased behavioral responses to ethanol, whereas increases in the levels of BDNF, mediated by the scaffolding protein RACK1, attenuate these behaviors. Interestingly, we found that acute exposure of neurons to ethanol leads to increased levels of BDNF mRNA via RACK1. Importantly, acute systemic administration of ethanol and voluntary ethanol consumption lead to increased levels of BDNF expression in the dorsal striatum. Taken together, these findings suggest that RACK1 and BDNF are part of a regulatory pathway that opposes adaptations that lead to the development of alcohol addiction. PMID- 15548670 TI - Dendrodendritic electrical synapses between mammalian retinal ganglion cells. AB - Electrical synapses between alpha-type ganglion cells were detected using combined techniques of dual patch-clamp recordings, intracellular labeling, electron microscopy, and channel subunit connexin immunocytochemistry in the albino rat retina. After intracellular injection of Neurobiotin into alpha-cells of inner (ON-center) and outer (OFF-center) ramifying types, measurement of tracer coupling resulted in a preferentially homologous occurrence among cells of the same morphological type (n = 19 of 24). In high-voltage as well as conventional electron microscopic analysis, direct dendrodendritic gap junctions (average size, 0.86 mum long) were present in contact sites between tracer coupled alpha-cells. In simultaneous dual whole-cell recordings from pairs of neighboring alpha-cells, these cells generated TTX-sensitive sustained spiking against extrinsic current injection, and bidirectional electrical synapses (maximum coupling coefficient, 0.32) with symmetrical junction conductance (average, 1.35 nS) were observed in pairs with cells of the same morphological type. Precise temporal synchronization of spike activity (average time delay, 2.7 msec) was detected when depolarizing currents were simultaneously injected into the pairs. To address whether physiologically identified electrical synapses constitute gap junctional connectivity between cell pairs, identified neuronal connexin36 immunoreactivity was undertaken in Lucifer yellow-labeled cell pairs after patch-clamp recordings. All alpha-cells expressed connexin36, and confocal laser-scanning imaging demonstrated that connexin36 is primarily located at dendritic crossings between electrically coupled cells (seven sites in a pair, on average). These results give conclusive evidence for electrical synapses via dendrodendritic gap junctions involving connexin36 in alpha retinal ganglion cells of the same physiological type. PMID- 15548671 TI - A genetic switch for epilepsy in adult mice. AB - Premature death from seizures afflicts gene-targeted mice expressing the Q/R site unedited glutamate receptor subunit GluR-B(Q) of AMPA receptors in central neurons. Early seizure-related death has now been circumvented by a genetic switch that restricts GluR-B(Q) expression to forebrain principal neurons from postnatal stages onward, prominently in hippocampus and striatum and less so in cortex and amygdala. When switched on, functional receptor incorporation of GluR B(Q) could be demonstrated by imaging evoked AMPA channel-mediated spinous Ca2+ transients in CA1 pyramidal cells. Sustained GluR-B(Q) expression in adult mice led to smaller excitatory postsynaptic responses in the CA1 region with unchanged presynaptic fiber excitability. Notably, despite the smaller excitatory response, the CA1 cells exhibited a reduced population spike threshold, which might underlie the spontaneous manifestations of epilepsy, including myocloni and generalized seizures with limbic components, observed by synchronous video monitoring and electroencephalographic recordings. No neuropathological symptoms developed when GluR-B(Q) expression was restricted to only hippocampal neurons. Our results show that seizure susceptibility is triggered by GluR-B(Q) expression also in the adult brain and that circuit hyperexcitability is not an immediate consequence of GluR-B(Q) but requires yet unknown downstream events, likely to be induced by non-Hebbian plasticity from Ca2+-permeable AMPA channels in principal neurons. PMID- 15548674 TI - Endogenous inhibitors of nuclear factor-kappaB, an opportunity for cancer control. AB - Excessive and prolonged activation of nuclear factor-kappaB (NF-kappaB) has been linked to numerous human diseases, especially cancer, because of the elevated expression of genes encoding antiapoptotic proteins, cytokines, chemokines, cell adhesion molecules, and so on. Eukaryotic cells have developed multiple mechanisms to keep this ubiquitous transcription factor in check. In addition to the inhibitor of kappaB family proteins, a number of endogenous molecules that negatively regulate the activation or activity of NF-kappaB have been identified. These molecules include A20, CYLD, cyPG15-deoxy-Delta(12,14)-prostaglandin J(2), Foxj1, Twist proteins, and beta-arrestins. The extended list of these endogenous inhibitors of NF-kappaB may provide new opportunities for the development of novel strategies for the intervention of malignant transformation. The question to be asked is how NF-kappaB is sustained activated in a number of cancers in which so many antagonists are surrounded. PMID- 15548675 TI - Enhancement of hypoxia-induced tumor cell death in vitro and radiation therapy in vivo by use of small interfering RNA targeted to hypoxia-inducible factor-1alpha. AB - Hypoxia-inducible factor-1alpha (HIF-1alpha) is an important transcriptional factor that is activated when mammalian cells experience hypoxia, a tumor microenvironmental condition that plays pivotal roles in tumor progression and treatment. In this study, we examined the idea of down-regulating HIF-1alpha in tumor cells for therapeutic gain. We show that the expression levels of HIF 1alpha can be significantly attenuated by use of the recently established small interfering RNA technology in combination with adenovirus-mediated gene transfer. Down-regulation of the HIF-1alpha protein enhanced hypoxia-mediated tumor cell apoptosis in vitro. Subcutaneous tumor growth was also prevented from cells with attenuated HIF-1alpha expression. In addition, intratumoral injection of adenovirus encoding the HIF-1alpha-targeted small interfering RNA had a small but significant effect on tumor growth when combined with ionizing radiation. Therefore, our results provide proof of HIF-1alpha as an effective target for anticancer therapy. They also suggest that an adenovirus-based small interfering RNA gene transfer approach may be a potentially effective adjuvant strategy for cancer treatment. PMID- 15548676 TI - Significant contribution of germline BRCA2 rearrangements in male breast cancer families. AB - Although screening for large deletions or duplications of the BRCA1 gene is becoming a routine component of the molecular diagnosis of familial breast cancer, little is known about the occurrence of such rearrangements in the BRCA2 gene. Because of the high frequency of BRCA2 mutations in breast cancer families with at least one case of male breast cancer, we selected a cohort of 39 such families, tested negative for mutations in the coding regions of BRCA1 and BRCA2, and developed an assay for BRCA2 rearrangements, based on quantitative multiplex PCR of short fluorescent fragments (QMPSF). We found three rearrangements: (1) a deletion of exons 12 and 13; (2) a duplication of exons 1 and 2; and (3) a complete deletion of BRCA2. We determined the boundaries of the deletion of exons 12 and 13, showing that it resulted from an unequal recombination between Alu sequences. We mapped the complete BRCA2 deletion, which extends over at least 298 kb and showed that it does not affect APRIN/AS3, previously characterized as a tumor suppressor gene, but it comprises several loci corresponding to proven or putative transcripts of unknown functional significance. These data suggest that screening for BRCA2 rearrangements should be done, especially in male breast cancer families tested negative for BRCA1 and BRCA2 mutations. PMID- 15548677 TI - Inhibition of extracellular-signal regulated kinases 1/2 is required for apoptosis of human colon cancer cells in vitro by sulindac metabolites. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) including sulindac have shown potent chemopreventive and tumor regressive effects against colorectal cancer, the second leading cause of cancer death in the United States. However, the mechanisms by which sulindac inhibits tumor cell growth are not completely understood. We previously reported that sulindac metabolites inhibit the mitogen activated protein/extracellular signal-regulated kinase kinase/extracellular signal-regulated kinase (MEK/ERK) signaling cascade in colorectal cancer cell lines at doses that induce apoptosis, and inhibition of MEK/ERK activity with U0126 is sufficient to induce apoptotic cell death. To determine whether inhibition of MEK/ERK activity is necessary for sulindac-induced apoptosis of human colon cancer cells, stable transfectants were created that express an activated MEK1 gene in HT29 cells. HT29-MEK1(R4F) clones displayed a 10- to 20 fold increase in MEK1 activity compared with control HT29-pCEP4 clones. When compared with control HT29-pCEP4 clones, HT29-MEK1(R4F) clones were resistant to both apoptosis and inhibition of ERK1/2 phosphorylation induced by sulindac metabolites. These results suggest that inhibition of MEK/ERK signaling is necessary for the induction of apoptosis by sulindac metabolites. PMID- 15548678 TI - Cul4A physically associates with MDM2 and participates in the proteolysis of p53. AB - The cullin 4A (Cul4A) gene is amplified and overexpressed in breast and hepatocellular carcinomas. Cul4A functions as an E3 ligase and participates in the proteolysis of several regulatory proteins through the ubiquitin-proteasome pathway. Here, we show that Cul4A associates with MDM2 and p53. Depletion of Cul4A leads to an accumulation of p53. Moreover, expression of Cul4A increases the decay-rate of p53 and delays the accumulation of p53 in response to DNA damage. Cul4A fails to increase the decay of p53 in mouse embryonic fibroblasts lacking MDM2. In addition, the Cul4A-mediated rapid decay of p53 is blocked by p19ARF. The results provide evidence for a role of Cul4A in the MDM2-mediated proteolysis of p53. PMID- 15548679 TI - Frequent aberrant methylation of the CDH4 gene promoter in human colorectal and gastric cancer. AB - Gene promoter methylation causes loss of tumor suppressor genes function in human cancer. Here, we show that the CDH4 gene, a member of the cadherin family encoding for R-cadherin, contains a CpG island located at the 5' of the first exon, which functions as a promoter element and is frequently affected by methylation in human cancer. By using methylation-specific PCR and reverse transcription-PCR in human cancer cell lines, promoter methylation could be directly linked to loss of gene expression. After treatment with the demethylating agent 5-aza-2-deoxycytidine, expression could be restored. Analysis of human primary tumors revealed that the CDH4 gene is methylated in 78% (38 of 49) of colorectal and 95% (20 of 21) of gastric carcinomas. CDH4 methylation was not detected in nonneoplastic colonic (0 of 10) and stomach (0 of 10) tissues or in peripheral blood (0 of 17). CDH4 methylation was detected in histologically normal tissues located in proximity of the neoplasms, indicating that CDH4 methylation is an early event in gastrointestinal tumor progression. We also proved that CDH4 methylation can be revealed in the peripheral blood of cancer patients. Our results indicate that CDH4 may act as a tumor suppressor gene in human gastrointestinal tumors and can potentially be used as an early diagnostic marker for gastrointestinal tumorigenesis. PMID- 15548680 TI - Overexpression of glycine-extended gastrin inhibits parietal cell loss and atrophy in the mouse stomach. AB - Recently we have reported synergistic effects between glycine-extended gastrin (G gly) and amidated gastrin-17 on acid secretion in short-term infusion studies. In the present study, we examined the long-term effect of G-gly on the atrophy promoting effects of amidated gastrin in the mouse stomach with or without Helicobacter infection. Transgenic mice overexpressing amidated gastrin (INS-GAS mice), G-gly (MTI/G-gly mice), and both peptides (INS-GAS/G-gly mice) were used for assessment of acid secretion and ulcer susceptibility and histologic examination and scoring of preneoplastic lesions in response to the 3 and 6 months Helicobacter felis (H. felis) infection. We found that MTI/G-gly mice had normal gastric histology and acid secretion. Double transgenic (INS-GAS/G-gly) mice showed 2-fold increases in acid secretion compared with INS-GAS mice. Acute peptic ulcers after pyloric ligation were noted in 50% of the INS-GAS/G-gly mice but in none of the INS-GAS mice at 6 months of age. Whereas male INS-GAS mice had a >50% decrease in the numbers of parietal cell and enterochromaffin-like cell at 6 months of age, the male double transgenic mice had no such decrease. Overexpression of G-gly reduced the scores of preneoplasia in the stomach; however, it did not prevent the development of amidated gastrin-dependent gastric cancer in both H. felis-infected mice and uninfected mice. We conclude that G-gly synergizes with amidated gastrin to stimulate acid secretion and inhibits parietal cell loss in INS-GAS/G-gly mice. The overexpression of G-gly seems to increase the susceptibility to peptic ulcer disease and delay the development of Helicobacter-mediated gastric preneoplasia in this model. PMID- 15548681 TI - Mechanistic and predictive profiling of 5-Fluorouracil resistance in human cancer cells. AB - Gene expression was analyzed in five pairs of 5-fluorouracil (5-FU) resistant and parental cancer cell lines on DNA microarrays. In unsupervised analysis, a prediction rule was built from the expression profiles of 29 genes, and 5-FU sensitivity class was predicted with 100% accuracy and high predictive strength. In supervised analysis of key 5-FU pathways, expression of 91 genes was associated with 5-FU sensitivity phenotype and segregated samples accordingly in hierarchical analysis. Key genes involved in 5-FU activation were significantly down-regulated (thymidine kinase, 2.9-fold; orotate phosphoribosyltransferase, 2.3-fold; uridine monophosphate kinase, 3.2-fold; pyrimidine nucleoside phosphorylase 3.6-fold) in resistant cells. Overexpression of thymidylate synthase and its adjacent gene, c-Yes, was detected in the resistant cell lines. The mRNA and protein overexpression of nuclear factor kappaB (NFkappaB) p65 and related antiapoptotic c-Flip gene was detected in resistant cells. The 5-FU resistant cell lines also showed high NFkappaB DNA-binding activity. Cotransfection of NFkappaB p50 and p65 cDNA induced 5-FU resistance in MCF-7 cells. Both NFkappaB- and 5-FU-induced resistant cell lines manifested reduced expression of genes governing G(1)-S and S-phase transition. Expression of genes involved in DNA replication was also down-regulated in resistant cell lines. These findings were highly consistent with the slower growth rate, higher proportion of G(1), and lower proportion of S-phase cells in the resistant cell lines. This phenotype may protect resistant cells from cell death induced by incorporation of 5-FU into DNA chains, by allowing time to repair 5-FU-induced damage. Our findings may provide novel targets for tackling 5-FU resistance. PMID- 15548682 TI - Characterization of a carcinogenesis rat model of ovarian preneoplasia and neoplasia. AB - Animal models of ovarian cancer are crucial for understanding the pathogenesis of the disease and for testing new treatment strategies. A model of ovarian carcinogenesis in the rat was modified and improved to yield ovarian preneoplastic and neoplastic lesions that pathogenetically resemble human ovarian cancer. A significantly lower dose (2 to 5 mug per ovary) of 7,12 dimethylbenz(a)anthracene (DMBA) was applied to the one ovary to maximally preserve its structural integrity. DMBA-induced mutagenesis was additionally combined with repetitive gonadotropin hormone stimulation to induce multiple cycles of active proliferation of the ovarian surface epithelium. Animals were treated in three arms of different doses of DMBA alone or followed by hormone administration. Comparison of the DMBA-treated ovaries with the contralateral control organs revealed the presence of epithelial cell origin lesions at morphologically distinct stages of preneoplasia and neoplasia. Their histopathology and path of dissemination to other organs are very similar to human ovarian cancer. Hormone cotreatment led to an increased lesion severity, indicating that gonadotropins may promote ovarian cancer progression. Point mutations in the Tp53 and Ki-Ras genes were detected that are also characteristic of human ovarian carcinomas. Additionally, an overexpression of estrogen and progesterone receptors was observed in preneoplastic and early neoplastic lesions, suggesting a role of these receptors in ovarian cancer development. These data indicate that this DMBA animal model gives rise to ovarian lesions that closely resemble human ovarian cancer and it is adequate for additional studies on the mechanisms of the disease and its clinical management. PMID- 15548683 TI - Loss of estrogen receptor signaling triggers epigenetic silencing of downstream targets in breast cancer. AB - Alterations in histones, chromatin-related proteins, and DNA methylation contribute to transcriptional silencing in cancer, but the sequence of these molecular events is not well understood. Here we demonstrate that on disruption of estrogen receptor (ER) alpha signaling by small interfering RNA, polycomb repressors and histone deacetylases are recruited to initiate stable repression of the progesterone receptor (PR) gene, a known ERalpha target, in breast cancer cells. The event is accompanied by acquired DNA methylation of the PR promoter, leaving a stable mark that can be inherited by cancer cell progeny. Reestablishing ERalpha signaling alone was not sufficient to reactivate the PR gene; reactivation of the PR gene also requires DNA demethylation. Methylation microarray analysis further showed that progressive DNA methylation occurs in multiple ERalpha targets in breast cancer genomes. The results imply, for the first time, the significance of epigenetic regulation on ERalpha target genes, providing new direction for research in this classical signaling pathway. PMID- 15548684 TI - Regulation of protein catabolism by muscle-specific and cytokine-inducible ubiquitin ligase E3alpha-II during cancer cachexia. AB - The progressive depletion of skeletal muscle is a hallmark of many types of advanced cancer and frequently is associated with debility, morbidity, and mortality. Muscle wasting is primarily mediated by the activation of the ubiquitin-proteasome system, which is responsible for degrading the bulk of intracellular proteins. E3 ubiquitin ligases control polyubiquitination, a rate limiting step in the ubiquitin-proteasome system, but their direct involvement in muscle protein catabolism in cancer remains obscure. Here, we report the full length cloning of E3alpha-II, a novel "N-end rule" ubiquitin ligase, and its functional involvement in cancer cachexia. E3alpha-II is highly enriched in skeletal muscle, and its expression is regulated by proinflammatory cytokines. In two different animal models of cancer cachexia, E3alpha-II was significantly induced at the onset and during the progression of muscle wasting. The E3alpha-II activation in skeletal muscle was accompanied by a sharp increase in protein ubiquitination, which could be blocked by arginine methylester, an E3alpha selective inhibitor. Treatment of myotubes with tumor necrosis factor alpha or interleukin 6 elicited marked increases in E3alpha-II but not E3alpha-I expression and ubiquitin conjugation activity in parallel. E3alpha-II transfection markedly accelerated ubiquitin conjugation to endogenous cellular proteins in muscle cultures. These findings show that E3alpha-II plays an important role in muscle protein catabolism during cancer cachexia and suggest that E3alpha-II is a potential therapeutic target for muscle wasting. PMID- 15548685 TI - Comparison of the effect of mutant and wild-type p53 on global gene expression. AB - The mechanisms for "gain-of-function" phenotypes produced by mutant p53s such as enhanced proliferation, resistance to transforming growth factor-beta-mediated growth suppression, and increased tumorigenesis are not known. One theory is that these phenotypes are caused by novel transcriptional regulatory events acquired by mutant p53s. Another explanation is that these effects are a result of an imbalance of functions caused by the retention of some of the wild-type transcriptional regulatory events in the context of a loss of other counterbalancing activities. An analysis of the ability of DNA-binding domain mutants A138P and R175H, and wild-type p53 to regulate the expression levels of 6.9 x 10(3) genes revealed that the mutants retained only <5% of the regulatory activities of the wild-type protein. A138P p53 exhibited mostly retained wild type regulatory activities and few acquired novel events. However, R175H p53 possessed an approximately equal number of wild-type regulatory events and novel activities. This is the first report that, after examination of the regulation of a large unfocused set of genes, provides data indicating that remaining wild-type transcriptional regulatory functions existing in the absence of counterbalancing activities as well as acquired novel events both contribute to the gain-of function phenotypes produced by mutant p53s. However, mutant p53s are likely to be distinct in terms of the extent to which each mechanism contributes to their gain-of-function phenotypes. PMID- 15548686 TI - Nuclear hormone receptor NR4A2 is involved in cell transformation and apoptosis. AB - HeLaHF cells are transformation revertants of cervical cancer HeLa cells and have lost anchorage-independent growth potential and tumorigenicity. Activation of tumor suppressor(s) was implicated previously in this transformation reversion. In this study, expression profiling analysis was carried out to identify potential oncogenes that are down-regulated in HeLaHF cells. We found that all three members of the NR4A1/Nur77/NGFIB orphan nuclear hormone receptor subfamily (NR4A1, NR4A2, and NR4A3) were down-regulated in the HeLaHF revertant. Small interfering RNA-mediated down-regulation of NR4A2 in HeLa cells, either transiently or stably, resulted in reduced anchorage-independent growth that was largely attributable to increased anoikis. Furthermore, down-regulation of NR4A2 as well as NR4A1 promoted intrinsic apoptosis. These phenotypes were also observed in several other experimental cancer cells, suggesting the observed apoptosis suppression is a more general property of NR4A2 and NR4A1. These phenotypes also suggest that the Nur77/NGFIB subfamily of orphan receptors exhibit certain oncogenic functionalities with regards to cell proliferation and apoptosis and could therefore be evaluated as potential cancer therapeutic targets. PMID- 15548687 TI - DNA microarrays reveal relationship of Ewing family tumors to both endothelial and fetal neural crest-derived cells and define novel targets. AB - Ewing family tumors (EFTs) are small round blue cell tumors that show features of neuroectodermal differentiation. However, the histogenetic origin of EFTs is still a matter of debate. We used high-density DNA microarrays for the identification of EFT-specific gene expression profiles in comparison with normal tissues of diverse origin. We identified 37 genes that are up-regulated in EFTs compared with normal tissues and validated expression of these genes in EFTs by both conventional and quantitative reverse transcription-polymerase chain reaction. The expression pattern of EFT-associated genes in normal tissues indicated a high similarity between EFTs and fetal and neuronal as well as endothelial tissues and supports the concept that a primitive neural crest derived progenitor at the transition to mesenchymal and endothelial differentiation is transformed in EFTs. EFT-associated genes could be used for molecular discrimination between EFTs and other small round blue cell tumors and clearly identified a cell line (SK-N-MC) that was initially established as neuroblastoma as being an EFT. Ectopic expression of the EFT-specific EWS-FLI1 fusion protein in human embryonic kidney (HEK293) cells was not sufficient to induce the complete EFT-specific gene expression signature, suggesting that the EFT-specific gene expression profile is not just a consequence of EWS-FLI1 expression but depends on the histogenetic background of the EFT stem cell. PMID- 15548688 TI - Inhibition of melanoma growth and metastasis by ATF2-derived peptides. AB - The resistance of melanoma to apoptosis, as well as its growth and metastasis capabilities, can be overcome by expression of a peptide derived from amino acid (aa) 51 to 100 of ATF2. Here we show that expression of ATF2((51-100)) in human melanoma cells reduced their growth in nude mice, which was additionally inhibited upon treatment with protein kinase inhibitors UCN-01 or SB203580. Injection of a fusion protein consisting of HIV-TAT and aa 51 to 100 of ATF2 into SW1 melanomas efficiently inhibits their growth and their metastasis up to complete regression. Additionally, expression of a 10aa peptide that corresponds to aa 51 to 60 of ATF2 sensitizes melanoma cells to spontaneous apoptosis, which coincides with activation of caspase 9 and poly(ADP-ribose) polymerase cleavage, and inhibit their growth in vivo. The 10aa peptide increases the association of c Jun NH(2)-terminal kinase with c-Jun but not with ATF2, resulting in concomitant increase in TRE-mediated transcription. Our study points to mechanisms underlying the activities of the ATF2 peptide while highlighting its possible use in drug design. PMID- 15548689 TI - Ionizing radiation induces frequent translocations with delayed replication and condensation. AB - Certain chromosome rearrangements display a significant delay in replication timing that is associated with a delay in mitotic chromosome condensation. Chromosomes with delay in replication timing/delay in mitotic chromosome condensation participate in frequent secondary rearrangements, indicating that cells with delay in replication timing/delay in mitotic chromosome condensation display chromosomal instability. In this report, we show that exposing cell lines or primary blood lymphocytes to ionizing radiation results in chromosomes with the delay in replication timing/delay in mitotic chromosome condensation phenotype, and that the delay in replication timing/delay in mitotic chromosome condensation phenotype occurs predominantly on chromosome translocations. In addition, exposing mice to ionizing radiation also induces cells with delay in replication timing/delay in mitotic chromosome condensation chromosomes that persist for as long as 2 years. Cells containing delay in replication timing/delay in mitotic chromosome condensation chromosomes frequently display hyperdiploid karyotypes, indicating that delay in replication timing/delay in mitotic chromosome condensation is associated with aneuploidy. Finally, using a chromosome engineering strategy, we show that only a subset of chromosome translocations displays delay in replication timing/delay in mitotic chromosome condensation. Our results indicate that specific chromosome rearrangements result in the generation of the delay in replication timing/delay in mitotic chromosome condensation phenotype and that this phenotype occurs frequently in cells exposed to ionizing radiation both in vitro and in vivo. PMID- 15548690 TI - Limited capability of regional lymph nodes to eradicate metastatic cancer cells. AB - The capacity of lymph nodes to eradicate cancer is a controversial issue. The purpose of this study was to determine the interplay between tumor growth and host resistance at early stages of lymph node metastasis. A metastasis model was made in the rat mesenteric lymph node, and migration of cancer cells was visualized in vivo. The lymph node was removed for histologic analysis and cytokine measurement. Migrant cancer cells were initially arrested in the marginal sinus. After an initial increase, the number of cancer cells in the marginal sinus declined until 48 hours after inoculation. Germinal centers and lymphoid cells in the medulla proliferated before 48 hours. ED3(+) macrophages incorporated apoptotic cancer cells, but significant cancer proliferation occurred after 4 days. Lymph nodes depleted of macrophages were massively invaded by cancer cells. Tumor necrosis factor alpha and interleukin (IL)-1beta in the nodes transiently increased after 1 hour and 3 hours, respectively, and were expressed in ED3(+) and ED2(+) macrophages, respectively. These changes were followed by a transient increase in IL-2. Interferon-gamma and IL-12 did not increase during the early stages of metastasis, but they decreased after 48 hours. In conclusion, the marginal sinus constitutes a mechanical barrier against cancer cell passage. Early pathological manifestations in the regional lymph node are consistent with those in cancer patients with improved survival. Parasinus macrophages play a role in the transient antimetastatic capability of the node, and cytokines secreted by these cells increased at the early stages of metastasis. Deterioration of cytokine induction may be responsible for subsequent cancer proliferation. PMID- 15548691 TI - Tumor-associated endothelial cells with cytogenetic abnormalities. AB - Tumor angiogenesis is necessary for solid tumor progression and metastasis. Tumor blood vessels have been shown to differ from normal counterparts, for example, by changes in morphology. An important concept in tumor angiogenesis is that tumor endothelial cells are assumed to be genetically normal, although these endothelial cells are structurally and functionally abnormal. However, we hypothesized that given the phenotypic differences between tumor and normal blood vessels, there may be genotypic alterations as well. Mouse endothelial cells were isolated from two different human tumor xenografts, melanoma and liposarcoma, and from two normal endothelial cell counterparts, skin and adipose. Tumor-associated endothelial cells expressed typical endothelial cell markers, such as CD31. They had relatively large, heterogeneous nuclei. Unexpectedly, tumor endothelial cells were cytogenetically abnormal. Fluorescence in situ hybridization (FISH) analysis showed that freshly isolated uncultured tumor endothelial cells were aneuploid and had abnormal multiple centrosomes. The degree of aneuploidy was exacerbated by passage in culture. Multicolor FISH indicated that the structural chromosomal aberrations in tumor endothelial cells were heterogeneous, indicating that the cytogenetic alterations were not clonal. There was no evidence of human tumor derived chromosomal material in the mouse tumor endothelial cells. In marked contrast, freshly isolated normal skin and adipose endothelial cells were diploid, had normal centrosomes, and remained cytogenetically stable in culture even up to 20 passages. FISH analysis of tumor sections also showed endothelial cell aneuploidy. We conclude that tumor endothelial cells can acquire cytogenetic abnormalities while in the tumor microenvironment. PMID- 15548692 TI - Physical and functional interactions between the Wwox tumor suppressor protein and the AP-2gamma transcription factor. AB - The WWOX gene encodes a tumor suppressor WW domain-containing protein, Wwox. Alterations of WWOX have been demonstrated in multiple types of cancer, and introduction of Wwox into Wwox-negative tumor cells has resulted in tumor suppression and apoptosis. The Wwox protein contains two WW domains that typically bind proline-rich motifs and mediate protein-protein interactions. Recently, we have described functional cross-talk between the Wwox protein and the p53 homologue, p73. To further explore the biological function of Wwox, we investigated other interacting candidates. In this report, we demonstrate a physical and functional association between AP-2gamma transcription factor and the Wwox protein. AP-2gamma at 20q13.2 encodes a transcription factor and is frequently amplified in breast carcinoma. We show that Wwox binds to the PPPY motif of AP-2gamma via its first WW domain. Alterations of tyrosine 33 in the first WW domain of Wwox or the proline-rich motif in AP-2gamma dramatically reduce this interaction. In addition, our results demonstrate that Wwox expression triggers redistribution of nuclear AP-2gamma to the cytoplasm, hence suppressing its transactivating function. Our results suggest that Wwox tumor suppressor protein inhibits AP-2gamma oncogenic activity by sequestering it in the cytoplasm. PMID- 15548693 TI - BRAK/CXCL14 is a potent inhibitor of angiogenesis and a chemotactic factor for immature dendritic cells. AB - BRAK/CXCL14 is a CXC chemokine constitutively expressed at the mRNA level in certain normal tissues but absent from many established tumor cell lines and human cancers. Although multiple investigators cloned BRAK, little is known regarding the physiologic function of BRAK or the reason for decreased expression in cancer. To understand the possible significance associated with loss of BRAK mRNA in tumors, we examined the pattern of BRAK protein expression in normal and tumor specimens from patients with squamous cell carcinoma (SCC) of the tongue and used recombinant BRAK (rBRAK) to investigate potential biological functions. Using a peptide-specific antiserum, abundant expression of BRAK protein was found in suprabasal layers of normal tongue mucosa but consistently was absent in tongue SCC. Consistent with previous in situ mRNA studies, BRAK protein also was expressed strongly by stromal cells adjacent to tumors. In the rat corneal micropocket assay, BRAK was a potent inhibitor of in vivo angiogenesis stimulated by multiple angiogenic factors, including interleukin 8, basic fibroblast growth factor, and vascular endothelial growth factor. In vitro, rBRAK blocked endothelial cell chemotaxis at concentrations as low as 1 nmol/L, suggesting this was a major mechanism for angiogenesis inhibition. Although only low affinity receptors for BRAK could be found on endothelial cells, human immature monocyte derived dendritic cells (iDCs) bound rBRAK with high affinity (i.e., K(d), approximately 2 nmol/L). Furthermore, rBRAK was chemotactic for iDCs at concentrations ranging from 1 to 10 nmol/L. Our findings support a hypothesis that loss of BRAK expression from tumors may facilitate neovascularization and possibly contributes to immunologic escape. PMID- 15548694 TI - Role of synaptojanin 2 in glioma cell migration and invasion. AB - The small GTPase Rac1 is thought to play an important role in cell migration and invasion. We have previously identified synaptojanin 2, a phosphoinositide phosphatase, as an effector of Rac1. Here, we show that small interfering RNA mediated depletion of either Rac1 or synaptojanin 2 inhibits invasion of SNB19 and U87MG glioblastoma cells through Matrigel and rat brain slices. Depletion of Rac1 or synaptojanin 2 also inhibits migration of SNB19 and U87MG cells on glioma derived extracellular matrix. In addition, we found that depletion of Rac1 or synaptojanin 2 inhibits the formation of lamellipodia and invadopodia, specialized membrane structures that are thought to be involved in extracellular matrix degradation. These results suggest that synaptojanin 2 contributes to the role of Rac1 in cell invasion and migration by regulating the formation of invadopodia and lamellipodia. This study also identifies synaptojanin 2 as a novel potential target for therapeutic intervention in malignant tumors. PMID- 15548695 TI - Bone morphogenetic protein (BMP)-6 signaling and BMP antagonist noggin in prostate cancer. AB - It has been proposed that the osteoblastic nature of prostate cancer skeletal metastases is due in part to elevated activity of bone morphogenetic proteins (BMPs). BMPs are osteoinductive morphogens, and elevated expression of BMP-6 correlates with skeletal metastases of prostate cancer. In this study, we investigated the expression levels of BMPs and their modulators in prostate, using microarray analysis of cell cultures and gene expression. Addition of exogenous BMP-6 to DU-145 prostate cancer cell cultures inhibited their growth by up-regulation of several cyclin-dependent kinase inhibitors such as p21/CIP, p18, and p19. Expression of noggin, a BMP antagonist, was significantly up-regulated by BMP-6 by microarray analysis and was confirmed by quantitative reverse transcription-polymerase chain reaction and at the protein level. Noggin protein was present in prostate biopsies and localized to the epithelial components of prostate by immunohistochemistry. Recombinant noggin inhibited the function of BMP-6, suggesting a negative feedback regulation of BMP activity and indicating a strategy for the development of a novel therapeutic target in the treatment of painful osteosclerotic bone metastases of prostate cancer. PMID- 15548696 TI - Host acid sphingomyelinase regulates microvascular function not tumor immunity. AB - Previous studies provided evidence that MCA/129 fibrosarcomas and B16 melanomas grow 2- to 4-fold faster in acid sphingomyelinase (asmase)-deficient mice than in asmase(+/+) littermates and are resistant to single-dose irradiation due to inability to mount an apoptotic response in tumor microvascular endothelium. However, others postulated the differences might be associated with a host antitumor immune response in asmase(+/+) mice that is not expressed in asmase(-/ ) mice due to phenotypic deficiency in antitumor immunity. The present studies demonstrate that none of the tumor-host combinations displayed the classic criteria of an immunogenic tumor because they lacked endotumoral or peritumoral infiltrates almost entirely. Furthermore, neither MCA/129 fibrosarcoma nor B16 melanoma tumors showed differences in growth or radioresponsiveness when implanted into mutant mouse models (Rag(-/-) and MEF(-/-)) lacking functional immune cell [natural killer (NK), NK-T, T, and B cells] populations. Additionally, development and function of B-, T-, and NK-cell populations in asmase(-/-) mice were normal, indistinguishable from their wild-type littermates. These data provide definitive evidence that MCA/129 fibrosarcomas and B16F1 melanomas do not elicit a host immune response in wild-type mice and that the asmase(-/-) phenotype is not deficient in antitumor immunity, supporting the notion that the patterns of tumors growth and radiation response are conditionally linked to the ability of the tumor endothelium to undergo ASMase mediated apoptosis. PMID- 15548697 TI - SHP-2-dependent mitogen-activated protein kinase activation regulates EGFRvIII but not wild-type epidermal growth factor receptor phosphorylation and glioblastoma cell survival. AB - In human glioblastomas, the most common mutation of epidermal growth factor receptor (EGFR) is an in-frame deletion of an 801-bp sequence in the extracellular domain of EGFR termed EGFRvIII. The EGFRvIII does not bind ligand EGF but has constitutive tyrosine phosphorylation (pTyr) content and kinase activity that result in enhanced transformation, reduced apoptosis, and resistance to therapy. Here we report that the protein tyrosine phosphatase SHP-2 modulates a mitogen-activated protein kinase (MAPK) kinase (MEK)-mediated signaling pathway that regulates EGFRvIII pTyr and cell survival in U87MG.EGFRvIII cells. Overexpression of the phosphatase-inactive form of SHP-2 inhibited EGFRvIII pTyr by decreasing MAPK phosphorylation. Consistent with this, we observed that the MEK inhibitor PD98059, but not the phosphatidylinositol 3' kinase inhibitor LY294002, inhibited EGFRvIII pTyr. Furthermore, constitutive EGFRvIII pTyr content observed in U87MG, LN229, and U373MG glioblastoma cells, but not in NR6.EGFRvIII fibroblasts, correlated with elevated MAPK levels in these cells. Interestingly, LY294002, but not PD98059, inhibited wild-type EGFR pTyr in response to EGF treatment in U87MG parental cells and in wild-type EGFR overexpressing U87MG cells. Inhibition of EGFRvIII pTyr by PD98059 was not observed to be phosphorylation site specific. However, LY294002 more specifically inhibited wild-type EGFR pTyr at residues Tyr(992) and Tyr(1068) in the COOH terminus. Treatment of U87MG.EGFRvIII cells with PD98059, but not LY294002, also resulted in increased cell death in response to cisplatin. Collectively, a distinct MEK-mediated pathway in human glioblastoma cells appears to differentially modulate EGFRvIII and wild-type EGFR pTyr, and inhibition of the MAPK pathway sensitizes EGFRvIII-containing human glioblastoma cells to cisplatin induced cell death. PMID- 15548698 TI - The nuclear receptor coactivator AIB1 mediates insulin-like growth factor I induced phenotypic changes in human breast cancer cells. AB - The nuclear receptor coactivator AIB1 (amplified in breast cancer 1) is overexpressed in human breast cancers and is required for estrogen signaling. However, the role of AIB1 in breast cancer etiology is not known. Here, we show that AIB1 is rate-limiting for insulin-like growth factor I (IGF-I)-dependent phenotypic changes and gene expression in human breast cancer cells. Reduction of endogenous AIB1 levels by small interfering RNA in MCF-7 breast cancer cells prevented IGF-I-stimulated anchorage-independent growth by reducing IGF-I dependent anti-anoikis. cDNA array and immunoblot analysis of gene expression revealed that reduction in AIB1 levels led to a significant decrease in the expression of several genes controlling the cell cycle and apoptosis. These AIB1 dependent changes were also observed in the presence of estrogen antagonist and were corroborated in the estrogen receptor-negative cell line MDA MB-231. AIB1 reduction decreased the expression of the IGF-I receptor and IRS-1 in MCF-7 but not in MDA MB-231 cells. IGF-I-stimulated activation of AKT was reduced by AIB1 small interfering RNA treatment, whereas mitogen-activated protein kinase (extracellular signal-regulated kinase 1/2) activation by IGF-I was unaffected. We conclude that AIB1 is required for IGF-I-induced proliferation, signaling, cell survival, and gene expression in human breast cancer cells, independent of its role in estrogen receptor signaling. PMID- 15548699 TI - P-cadherin is up-regulated by the antiestrogen ICI 182,780 and promotes invasion of human breast cancer cells. AB - P-cadherin expression in breast carcinomas has been associated with tumors of high histologic grade and lacking estrogen receptor-alpha, suggesting a link between these proteins. In the MCF-7/AZ breast cancer cell line, blocking estrogen receptor-alpha signaling with the antiestrogen ICI 182,780 induced an increase of P-cadherin, which coincided with induction of in vitro invasion. Retroviral transduction of MCF-7/AZ cells, as well as HEK 293T cells, showed the proinvasive activity of P-cadherin, which requires the juxtamembrane domain of its cytoplasmic tail. This study establishes a direct link between P-cadherin expression and the lack of estrogen receptor-alpha signaling in breast cancer cells and suggests a role for P-cadherin in invasion, through its interaction with proteins bound to the juxtamembrane domain. PMID- 15548700 TI - Transactivation of the EGR1 gene contributes to mutant p53 gain of function. AB - Tumor-associated mutants of the p53 tumor suppressor protein exert biological activities compatible with an oncogenic gain of function. To explore the underlying molecular mechanism, we performed microarray analysis, comparing p53 null cells to mutant p53-expressing cells. One of the genes up-regulated in the presence of mutant p53 was EGR1, a transcription factor implicated in growth control, apoptosis, and cancer. EGR1 induction by various types of stress is markedly augmented in cells expressing mutant p53. Moreover, chromatin immunoprecipitation analysis indicates that mutant p53 is physically associated with the EGR1 promoter. Functional assays indicate that induction of EGR1 by mutant p53 contributes to enhanced transformed properties and resistance to apoptosis. We propose that EGR1 is a significant contributor to mutant p53 gain of function. PMID- 15548701 TI - 24-hour oscillation of mouse methionine aminopeptidase2, a regulator of tumor progression, is regulated by clock gene proteins. AB - Methionine aminopeptidase2 (MetAP2) plays an important role in the growth of endothelial cells during the tumor angiogenesis stage. Recently, we have clarified that mouse methionine aminopeptidases (mMetAPs) show a 24-hour rhythm in implanted tumor masses. In the present study, we investigated the mechanism underlying the 24-hour rhythm of mMetAP2 activity in tumor-bearing mice under a light-dark (lights on from 7 a.m. to 7 p.m.) cycle. The 5' flanking region of mMetAP2 included eight E-boxes. The transcription of the mMetAP2 promoter was enhanced by the mCLOCK:mBMAL1 heterodimer, and its activation was inhibited by mPER2 or mCRY1. Deletion and mutation of the E-boxes in the region indicated that the E-box nearest to the initiation start site played an important role in the transcriptional regulation by clock genes. In sarcoma180-bearing mice, the pattern of binding of mCLOCK and mBMAL1 to the E-box and transcription of the mMetAP2 promoter showed a 24-hour rhythm with higher levels from the mid-light to early dark phase. The pattern of mMetAP2 transcription was closely associated with that of mMetAP2 mRNA expression in three types of tumor-bearing mice. mMetAP2 protein expression varied with higher levels from the late-dark to early light phase. The rhythmicity of the protein expression was synchronous with that of the activity of mMetAPs but out of phase with that of the mMetAP2 mRNA expression. These results suggest that the 24-hour rhythm of mMetAP2 activity is regulated by the transcription of clock genes within the clock feedback loops. PMID- 15548702 TI - Nuclear betaII-tubulin associates with the activated notch receptor to modulate notch signaling. AB - The Notch signal pathway plays important roles in proliferation, apoptosis, and differentiation. Abnormalities in Notch signaling are linked to many human diseases. After ligand binding, Notch signaling is activated through the cleavage of Notch receptors to release and translocate the Notch intracellular domain into the nucleus. The Notch1 receptor intracellular domain (N1IC), the activated form of the Notch1 receptor, can modulate downstream target genes via C promoter binding factor 1-dependent and -independent pathways. To further dissect the Notch1 signaling pathway, we screened the N1IC-associated proteins using a yeast two-hybrid system and identified nuclear beta(II)-tubulin as a candidate for the N1IC-associated proteins. It was suggested that the presence of beta(II)-tubulin in nuclei might be correlated with the cancerous state of cells. However, the function of beta(II)-tubulin locating in the nucleus still is unknown. Herein, we show that the complex of alpha- and beta(II)-tubulin is associated with N1IC in cancer cells by a coimmunoprecipitation analysis. The ankyrin domain of the Notch1 receptor alone was sufficient to associate with beta(II)-tubulin. Furthermore, alpha- and beta(II)-tubulin were localized in the nucleus and formed a complex with N1IC. Treatment with Taxol increased the amounts of nuclear alpha- and beta(II)-tubulin in K562 and HeLa cells and promoted the C promoter-binding factor 1-dependent transactivation activity of N1IC. We also show that nuclear beta(II)-tubulin was bound on the C promoter-binding factor 1 response elements via the association with N1IC. These results suggest that nuclear beta(II) tubulin can modulate Notch signaling through interaction with N1IC in cancer cells. PMID- 15548703 TI - Lymphocyte transformation by Pim-2 is dependent on nuclear factor-kappaB activation. AB - Pim-2 is a transcriptionally regulated oncogenic kinase that promotes cell survival in response to a wide variety of proliferative signals. Deregulation of Pim-2 expression has been documented in several human malignancies, including leukemia, lymphoma, and multiple myeloma. Here, we show that the ability of Pim-2 to promote survival of cells is dependent on nuclear factor (NF)-kappaB activation. Pim-2 activates NF-kappaB-dependent gene expression by inducing phosphorylation of the oncogenic serine/threonine kinase Cot, leading to both augmentation of IkappaB kinase activity and a shift in nuclear NF-kappaB from predominantly p50 homodimers to p50/p65 heterodimers. Blockade of NF-kappaB function eliminates Pim-2-mediated survival in both cell lines and primary cells, and both Cot phosphorylation and expression are required for the prosurvival effects of Pim-2. Although Pim-2 cooperates with Myc to promote growth factor independent cell proliferation, this feature is abrogated by NF-kappaB blockade. The ability of Pim-2 to serve as an oncogene in vivo depends on sustained NF kappaB activity. Thus, the transcriptional induction of Pim-2 initiates a novel NF-kappaB activation pathway that regulates cell survival. PMID- 15548704 TI - Interferon gamma enhances the effectiveness of tumor necrosis factor-related apoptosis-inducing ligand receptor agonists in a xenograft model of Ewing's sarcoma. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces selective apoptosis in a variety of tumors, including most cell lines derived from Ewing's sarcoma family of tumors, an aggressive sarcoma that afflicts children and young adults. To determine the in vivo efficacy of TRAIL receptor agonists in Ewing's sarcoma family of tumors, mice with orthotopic xenografts were treated with anti TRAIL-R2 monoclonal antibody or TRAIL/Apo2L in a model that can identify effects on both primary tumors and metastases. Administration of either agonist slowed tumor growth in 60% of animals and induced durable remissions in 11 to 19% but did not alter the incidence of metastatic disease. Response rates were not improved by concurrent doxorubicin treatment. Cells recovered from both TRAIL receptor agonist-treated and nontreated tumors were found to be resistant to TRAIL-induced death in vitro unless pretreated with interferon (IFN) gamma. This resistance coincided with a selective down-regulation of TRAIL receptor expression on tumor cells. In vivo treatment with IFNgamma increased tumor expression of TRAIL receptors and caspase 8, but did not increase the antitumor effect of TRAIL receptor agonists on primary tumors. However, IFNgamma treatment alone or in combination with a TRAIL receptor agonist significantly decreased the incidence of metastatic disease and the combination of TRAIL receptor agonist therapy with IFNgamma-mediated impressive effects on both primary tumors and metastatic disease. These data demonstrate that in vivo growth favors TRAIL resistance but that TRAIL receptor agonists are active in Ewing's sarcoma family of tumors and that the combination of TRAIL receptor agonists with IFNgamma is a potent regimen in this disease capable of controlling both primary and metastatic tumors. PMID- 15548705 TI - STAT1: a modulator of chemotherapy-induced apoptosis. AB - The anthracyclines, such as doxorubicin, are widely used in the treatment of breast cancer. Previously, we showed that these drugs could activate the transcription factor, nuclear factor kappaB, in a DNA damage-dependent manner. We now show that these drugs can potentiate the activation of signal transducer and activator of transcription 1 (STAT1) in MDA-MB 435 breast cancer cells treated with IFN-gamma. We observed that key markers of STAT1 activation, including tyrosine 701 and serine 727 phosphorylation, were enhanced in the presence of doxorubicin. This potentiation resulted in enhanced nuclear localization of activated STAT1 and led to an increase in the nuclear binding of activated STAT complexes. The observed potentiation was specific for STAT1 and IFN-gamma, as no effects were observed with either STAT3 or STAT5. Furthermore, the type I IFNs (alpha and beta) had little or no effect. The observed effects on STAT1 phosphorylation have previously been linked with maximal transcriptional activation and apoptosis. Cell viability was assessed by crystal violet staining followed by analysis with CalcuSyn to determine combination index values, a measure of synergy. We confirmed that significant synergy existed between IFN gamma and doxorubicin (combination index = 0.34) at doses lower than IC(50) values for this drug (0.67 micromol/L). In support of this, we observed that apoptotic cell death was also enhanced by measuring poly(ADP-ribose) polymerase and caspase-3 cleavage. Finally, suppression of STAT1 expression by small interfering RNA resulted in a loss of synergistic apoptotic cell death compared with cells, where no suppression of STAT1 expression was attained with scrambled small-interfering RNA control. We conclude that doxorubicin potentiates STAT1 activation in response to IFN-gamma, and that this combination results in enhanced apoptosis in breast cancer cells. PMID- 15548706 TI - Evidence that TRPM8 is an androgen-dependent Ca2+ channel required for the survival of prostate cancer cells. AB - The Ca(2+)-permeable channel TRPM8 is thought to play an important role in the pathophysiology of prostate cancer. We have investigated the intracellular location of TRPM8 and its role as a Ca(2+)-permeable channel in an androgen responsive and an androgen-insensitive prostate cancer cell line. We report evidence from immunofluorescence experiments that in the androgen-responsive LNCaP cell line, the TRPM8 protein is expressed in the endoplasmic reticulum and plasma membrane, acts as a Ca(2+)-permeable channel (assessed using Fura-2 to measure increases in the cytoplasmic Ca(2+) concentration) in each of these membranes, and is regulated by androgen. Although TRPM8 was detected in the androgen-insensitive PC-3 cell line, no evidence was obtained for regulation of its expression by androgen. The results of experiments using LNCaP cells, the TRPM8 antagonist capsazepine, and small interference RNA targeted to TRPM8 indicate that TRPM8 is required for cell survival. These results indicate that TRPM8 is an important determinator of Ca(2+) homeostasis in prostate epithelial cells and may be a potential target for the action of drugs in the management of prostate cancer. PMID- 15548707 TI - The anticancer drug ellipticine forms covalent DNA adducts, mediated by human cytochromes P450, through metabolism to 13-hydroxyellipticine and ellipticine N2 oxide. AB - Ellipticine is an antineoplastic agent, the mode of action of which is considered to be based on DNA intercalation and inhibition of topoisomerase II. We found that ellipticine also forms the cytochrome P450 (CYP)-mediated covalent DNA adducts. We now identified the ellipticine metabolites formed by human CYPs and elucidated the metabolites responsible for DNA binding. The 7-hydroxyellipticine, 9-hydroxyellipticine, 12-hydroxyellipticine, 13-hydroxyellipticine, and ellipticine N(2)-oxide are generated by hepatic microsomes from eight human donors. The role of specific CYPs in the oxidation of ellipticine and the role of the ellipticine metabolites in the formation of DNA adducts were investigated by correlating the levels of metabolites formed in each microsomal sample with CYP activities and with the levels of the ellipticine-derived deoxyguanosine adducts in DNA. On the basis of this analysis, formation of 9-hydroxyellipticine and 7 hydroxyellipticine was attributable to CYP1A1/2, whereas production of 13 hydroxyellipticine and ellipticine N(2)-oxide, the metabolites responsible for formation of two major DNA adducts, was attributable to CYP3A4. Using recombinant human enzymes, oxidation of ellipticine to 9-hydroxyellipticine and 7 hydroxyellipticine by CYP1A1/2 and to 13-hydroxyellipticine and N(2)-oxide by CYP3A4 was corroborated. Homologue modeling and docking of ellipticine to the CYP3A4 active center was used to explain the predominance of ellipticine oxidation by CYP3A4 to 13-hydroxyellipticine and N(2)-oxide. PMID- 15548708 TI - Ectopic expression of interferon regulatory factor-1 promotes human breast cancer cell death and results in reduced expression of survivin. AB - The overexpression of the inhibitor of apoptosis protein, survivin, may provide tumor cells with a distinct survival advantage in situ; hence, therapeutic strategies have been designed to inhibit its expression. In this study, we ectopically expressed the interferon regulatory factor (IRF)-1 protein in the breast carcinoma cell lines MDA-MB-468 and SK-BR-3 using a recombinant adenovirus (Ad-IRF-1). By screening microarray analysis of cDNA from the human breast cancer cell line MDA-MB-468 infected with Ad-IRF-1, we observed a 15-fold down regulation of the survivin gene when compared with uninfected cells. Consequently, we tested survivin expression in Ad-IRF-1-infected MDA-MB-468 and SK-BR-3 breast cancer cell lines. Immunoblotting analyses supported the contention that ectopic expression of the IRF-1 protein results in down regulation of survivin protein expression that is independent of p53. In addition, Ad-IRF-1 infection of these human breast cancer cell lines induces the expression of p21. We also report that increased apoptosis is observed in tumor cells infected with Ad-IRF-1 compared with Ad-Psi5 mock-infected cells and that cell death is further augmented when the IRF-1-infected cells are cultured with Adriamycin. Moreover, in a xenogeneic mouse model of breast carcinoma, in vivo treatment of tumor-bearing mice with intratumoral Ad-IRF-1 injections results in tumor growth inhibition. In resected tumors from mice that had been treated with Ad-IRF-1, tumor cells that express the IRF-1 transgene have a predominant IRF-1 positive, survivin-negative phenotype. Collectively, these observations suggest that therapies designed to enhance IRF-1 expression within tumor cells may represent novel treatment strategies for breast cancer. PMID- 15548709 TI - Cyclin B1 is a critical target of RhoB in the cell suicide program triggered by farnesyl transferase inhibition. AB - Farnesyl transferase inhibitors (FTIs) have displayed limited efficacy in clinical trials, possibly because of their relatively limited cytotoxic effects against most human cancer cells. Therefore, efforts to leverage the clinical utility of FTIs may benefit from learning how these agents elicit p53-independent apoptosis in mouse models of cancer. Knockout mouse studies have established that gain of the geranylgeranylated isoform of the small GTPase RhoB is essential for FTI to trigger apoptosis. Here we demonstrate that Cyclin B1 is a crucial target for suppression by RhoB in this death program. Steady-state levels of Cyclin B1 and its associated kinase Cdk1 were suppressed in a RhoB-dependent manner in cells fated to undergo FTI-induced apoptosis. These events were not derivative of cell cycle arrest, because they did not occur in cells fated to undergo FTI induced growth inhibition. Mechanistic investigations indicated that RhoB mediated transcriptional suppression but also accumulation of Cyclin B1 in the cytosol at early times after FTI treatment, at a time before the subsequent reduction in steady-state protein levels. Enforcing Cyclin B1 expression attenuated apoptosis but not growth inhibition triggered by FTI. Moreover, enforcing Cyclin B1 abolished FTI antitumor activity in graft assays. These findings suggest that Cyclin B1 suppression is a critical step in the mechanism by which FTI triggers apoptosis and robust antitumor efficacy. Our findings suggest that Cyclin B1 suppression may predict favorable clinical responses to FTI, based on cytotoxic susceptibility, and they suggest a rational strategy to address FTI nonresponders by coinhibition of Cdk1 activity. PMID- 15548710 TI - Phosphatidylinositol 3'-kinase activation leads to multidrug resistance protein-1 expression and subsequent chemoresistance in advanced prostate cancer cells. AB - The development of androgen-independent growth in advanced carcinoma of the prostate (CaP) is associated with poor prognosis and few therapeutic options. Chemotherapeutic drugs offer the afflicted patient palliative benefits, but these are short-lived because of the chemoresistant nature of hormone-refractory prostate cancer. Given the high percentage of CaP patients with mutations in the PTEN tumor suppressor gene, we sought to determine the involvement of the phosphatidylinositol 3'-kinase (PI3K) cascade in the development of CaP drug resistance. PTEN-negative PC3 cells were observed to have increased resistance to both doxorubicin and paclitaxel when compared with PTEN-positive DU145 cells. Furthermore, modulation of PI3K activity with the use of constitutively active and dominant-negative inhibitors was found to affect the ability to CaP cells to respond to chemotoxic treatments. Additionally, inhibition of PI3K with a small molecular weight inhibitor (LY294002) was able to potentiate the antineoplastic activity of both doxorubicin and paclitaxel in CaP cells. Interestingly, multidrug resistance protein-1 (MRP-1) expression, but not MDR-1 (p glycoprotein), was observed to be induced as a consequence of PI3K activation in these cell types. Inhibition of MRP-1 expression via siRNA was observed to synergistically sensitize CaP cells to chemotoxic drugs while having no appreciable effect on cell growth in the absence of these compounds. Taken together, these data suggest that PI3K activation can lead to the development of chemoresistant cells in prostatic carcinomas through the up-regulation of MRP-1. Thus, inhibition of PI3K activity with concomitant administration of chemotoxic compounds may prove beneficial in preventing the development of drug resistance in patients with hormone-refractory prostate cancer. PMID- 15548711 TI - Synergistic interaction between sphingomyelin and gemcitabine potentiates ceramide-mediated apoptosis in pancreatic cancer. AB - We have examined the mechanism by which sphingomyelin (SM) enhances chemotherapy in human pancreatic cancer cells, focusing on the correlation between ceramide metabolism and apoptosis. Dose response curves for gemcitabine in the absence or presence of 0.2 mg/mL SM provided IC(50) values of 78.3 +/- 13.7 and 13.0 +/- 3.0 nmol/L, respectively. The cytotoxic effect of the combined treatment was synergistic (combination index = 0.36). Using annexin-V staining, the percentage of apoptotic cells was 3.6 +/- 2.6% for the untreated cells, 6.5 +/- 3.8% for the 0.2 mg/mL SM-treated cells, and 19.9 +/- 12.9% for the 100 nmol/L gemcitabine treated cells, but increased significantly to 42.1 +/- 12.7% with the combined treatment (P < 0.001, compared with gemcitabine-treated group). The percentage of cells losing mitochondrial membrane potential followed a similar trend. The ceramide content of untreated and gemcitabine-treated cells was not significantly different (0.46 +/- 0.29 and 0.59 +/- 0.34 pmol ceramide/nmole PO(4)). However, when 0.2 mg/mL SM was added, ceramide levels were 1.09 +/- 0.42 and 1.58 +/- 0.55 pmol ceramide/nmol PO(4), for the SM alone and SM with gemcitabine-treated cells, respectively (P = 0.038). Acidic SMase was activated by exposure to gemcitabine but not SM, whereas the activities of neutral SMase and glycosylceramide synthase did not change with either gemcitabine or SM. The data are consistent with gemcitabine-induced activation of acidic SMase and indicate that the addition of SM can yield increased production of ceramide, mitochondrial depolarization, apoptosis, and cell death. Because SM by itself is relatively nontoxic, addition of this lipid to agents that induce apoptosis may prove useful to enhance apoptosis and increase cytotoxicity in cancer cells. PMID- 15548712 TI - Anti-CD137 monoclonal antibody administration augments the antitumor efficacy of dendritic cell-based vaccines. AB - In weakly and poorly immunogenic tumor models, we examined the effects of stimulating CD137 (4-1BB) in vivo by administering anti-CD137 monoclonal antibody after tumor lysate-pulsed dendritic cell (TP-DC) vaccination. TP-DC subcutaneous vaccination induced a transient up-regulation of CD137 on T cells and natural killer (NK) cells within vaccine-primed lymph nodes (VPLNs). In established pulmonary and subcutaneous tumor models, anti-CD137 synergistically enhanced tumor regression after TP-DC vaccination. In the subcutaneous tumor model, the combined therapy resulted in improved survival. Combined therapy also resulted in improved local control of subcutaneous tumor after surgical resection. Anti-CD137 polarized the cytokine release of VPLNs and spleen cells in response to tumor antigen toward a type 1 (interferon-gamma) versus a type 2 (interleukin-4) profile. Cell depletion and the use of knockout animals identified that CD8(+), CD4(+), and NK cells were involved in the tumor rejection response and that CD8(+) cells had the major effector role. Anti-CD137 administration resulted in increased proliferation of adoptively transferred OT-1 CD8(+) T cells in the VPLNs of mice inoculated with B16-OVA TP-DCs. Polarization toward type 1 (interferon-gamma) versus type 2 (interleukin-4) was also observed with the OT-1 cells from VPLNs and spleen cells after anti-CD137 injections. This polarization effect was abrogated by the in vivo depletion of NK cells. These findings indicate that the adjuvant effect of anti-CD137 given in conjunction with TP-DC vaccination is associated with the polarization of T effector cells toward a type 1 response to tumor antigen and is mediated via NK cells. PMID- 15548713 TI - Increased survival, proliferation, and migration in metastatic human pancreatic tumor cells expressing functional CXCR4. AB - In this study, we have evaluated 11 pancreatic tumor cell lines and tumor cells from surgical samples of patients with pancreatic adenocarcinoma for expression of the chemokine receptor CXCR4. Six of 11 cell lines expressed detectable mRNA of CXCR4, with three cell lines (AsPC1, Capan1, and Hs766T) having substantial amounts of transcripts. Expression was higher in lines derived from metastatic lesions compared with those derived from primary tumors. Different inflammatory cytokines did not modify expression, whereas IFN-gamma down-regulated and hypoxia up-regulated CXCR4 transcripts. Transcript expression was associated with surface expression in pancreatic carcinoma cell lines. All surgical carcinoma samples tested expressed higher levels of CXCR4 than normal pancreatic ducts, which were used as reference tissue. The chemokine CXCL12 induced chemotaxis in CXCR4 positive pancreatic carcinoma cell lines, which was inhibited by anti-CXCR4 monoclonal antibody and by the antagonist AMD3100. Transendothelial migration, Matrigel invasion, and activation of matrix metalloproteases were also enhanced by CXCL12. In CXCR4-positive cell lines, CXCL12 stimulated cell proliferation. The cell line Hs766T produces high levels of CXCL12, and addition of the CXCR4 antagonist AMD3100 partially inhibited proliferation, indicating an autocrine loop. Moreover, the addition of exogenous CXCL12 inhibited apoptosis induced by serum starvation. These results indicate that the CXCR4 receptor is frequently expressed in metastatic pancreatic tumor cells. CXCR4 not only stimulates cell motility and invasion but also promotes survival and proliferation. Strategies to target CXCR4 expressed on tumor cells may be of benefit in patients with pancreatic cancer. PMID- 15548714 TI - Immunological prevention of a multigene cancer syndrome. AB - Vaccines effectively prevent the onset of tumors in transgenic mice carrying activated oncogenes; however, human tumors are caused by combined alterations in oncogenes and oncosuppressor genes. We evaluated the impact of prophylactic vaccines in HER-2/neu transgenic, p53 wild-type/null mice that succumb to an aggressive cancer syndrome comprising mammary and salivary gland carcinomas and rhabdomyosarcoma. A vaccine made of allogeneic mammary carcinoma cells expressing HER-2/neu and interleukin 12 afforded long-term protection from tumor onset. Tumor prevention was mediated by T cell-derived cytokines, in particular gamma interferon, and by anti-HER-2/neu antibodies. HER-2/neu expression was inhibited in target tissues of vaccinated mice, and somatic loss of the wild-type p53 allele did not occur. A highly effective vaccine against a single oncoprotein induced a powerful immune response that arrested multistep carcinogenesis in distinct target tissues. PMID- 15548715 TI - Highly successful therapeutic vaccinations combining dendritic cells and tumor cells secreting granulocyte macrophage colony-stimulating factor. AB - In an attempt to induce potent immune antitumor activities, we investigated, within the rat 9L gliosarcoma model, distal therapeutic vaccinations associating three therapies: dendritic cell vaccination, intratumoral granulocyte macrophage colony-stimulating factor (GM-CSF) gene transfer, and tumor apoptosis induction. Vaccines of dendritic cells coinjected with processed GM-CSF secreting 9L cells induced systemic responses, resulting in the complete regression of distant preimplanted 9L tumor masses in, with the best strategy, 94% of male rats. All of the cured rats developed a long-term resistance to a rechallenge with parental cells. The curative responses were correlated with the detection of elevated specific cytotoxic activities and a CD4+, CD8+ T cell-, and natural killer (NK) cell-mediated IFN-gamma production. The survival rate of the rat seemed more directly linked to the amount of GM-CSF secreted by the transduced tumor cells, which in turn depended on the toxicity of the apoptosis-inducing treatment, than to the level of apoptosis induced. Unexpectedly, alive GM-CSF secreting 9L cells became apoptotic when injected in vivo. Thus we documented the positive role of apoptosis in the induction of therapeutic antitumor responses by comparing, at equal GM-CSF exogenous supply, the effects of dendritic cells coinjected with apoptotic or necrotic 9L cells. The data showed the superior therapeutic efficiency of combined vaccines containing apoptotic tumor cells. In conclusion, vaccinations with dendritic cells associated with apoptotic tumor cells secreting GM-CSF show a very high therapeutic potency that should show promise for the treatment of human cancer. PMID- 15548716 TI - C-type lectin-like molecule-1: a novel myeloid cell surface marker associated with acute myeloid leukemia. AB - Acute myeloid leukemia (AML) has a poor prognosis due to treatment-resistant relapses. A humanized anti-CD33 antibody (Mylotarg) showed a limited response rate in relapsed AML. To discover novel AML antibody targets, we selected a panel of single chain Fv fragments using phage display technology combined with flow cytometry on AML tumor samples. One selected single chain Fv fragment broadly reacted with AML samples and with myeloid cell lineages within peripheral blood. Expression cloning identified the antigen recognized as C-type lectin-like molecule-1 (CLL-1), a previously undescribed transmembrane glycoprotein. CLL-1 expression was analyzed with a human anti-CLL-1 antibody that was generated from the single chain Fv fragment. CLL-1 is restricted to the hematopoietic lineage, in particular to myeloid cells present in peripheral blood and bone marrow. CLL-1 is absent on uncommitted CD34(+)/CD38(-) or CD34(+)/CD33(-) stem cells and present on subsets of CD34(+)/CD38(+) or CD34(+)/CD33(+) progenitor cells. CLL-1 is not expressed in any other tissue. In contrast, analysis of primary AMLs demonstrated CLL-1 expression in 92% (68 of 74) of the samples. As an AML marker, CLL-1 was able to complement CD33, because 67% (8 of 12) of the CD33(-) AMLs expressed CLL-1. CLL-1 showed variable expression (10-60%) in CD34(+) cells in chronic myelogenous leukemia and myelodysplastic syndrome but was absent in 12 of 13 cases of acute lymphoblastic leukemia. The AML reactivity combined with the restricted expression on normal cells identifies CLL-1 as a novel potential target for AML treatment. PMID- 15548717 TI - Bone marrow is a reservoir for CD4+CD25+ regulatory T cells that traffic through CXCL12/CXCR4 signals. AB - CD4(+)CD25(+) regulatory T cells (Tregs) mediate peripheral T-cell homeostasis and contribute to self-tolerance. Their homeostatic and pathologic trafficking is poorly understood. Under homeostatic conditions, we show a relatively high prevalence of functional Tregs in human bone marrow. Bone marrow strongly expresses functional stromal-derived factor (CXCL12), the ligand for CXCR4. Human Tregs traffic to and are retained in bone marrow through CXCR4/CXCL12 signals as shown in chimeric nonobese diabetic/severe combined immunodeficient mice. Granulocyte colony-stimulating factor (G-CSF) reduces human bone marrow CXCL12 expression in vivo, associated with mobilization of marrow Tregs to peripheral blood in human volunteers. These findings show a mechanism for homeostatic Treg trafficking and indicate that bone marrow is a significant reservoir for Tregs. These data also suggest a novel mechanism explaining reduced acute graft-versus host disease and improvement in autoimmune diseases following G-CSF treatment. PMID- 15548718 TI - Local administration of granulocyte/macrophage colony-stimulating factor increases the number and activation state of dendritic cells in the sentinel lymph node of early-stage melanoma. AB - The initial tumor-draining lymph node, the sentinel lymph node, not only constitutes the first expected site of micrometastasis but also the first point of contact between tumor-associated antigens and the adaptive immune system. A tumor-induced decrease in the frequency and activation state of sentinel lymph node dendritic cells will impair the generation of effective antitumor T-cell responses and increase the likelihood of metastatic spread. Here, we demonstrate that intradermal administration of granulocyte macrophage-colony stimulating factor around the excision site of stage I primary melanoma tumors increases the number and activation state of dendritic cells in the paracortical areas of the sentinel lymph node and enhances their binding to T cells. We conclude that local treatment of melanoma patients with granulocyte macrophage-colony stimulating factor, before surgery, conditions the sentinel lymph node microenvironment to enhance mature dendritic cell recruitment and hypothesize that this may be more conducive to the generation of T-cell-mediated antitumor immunity. PMID- 15548719 TI - CYP3A4, CYP3A5, and CYP3A43 genotypes and haplotypes in the etiology and severity of prostate cancer. AB - The CYP3A genes reside on chromosome 7q21 in a multigene cluster. The enzyme products of CYP3A4 and CYP3A43 are involved in testosterone metabolism. CYP3A4 and CYP3A5 have been associated previously with prostate cancer occurrence and severity. To comprehensively examine the effects of these genes on prostate cancer occurrence and severity, we studied 622 incident prostate cancer cases and 396 controls. Substantial and race-specific linkage disequilibrium was observed between CYP3A4 and CYP3A5 in both races but not between other pairs of loci. We found no association of CYP3A5 genotypes with prostate cancer or disease severity. CYP3A43*3 was associated with family history-positive prostate cancer (age- and race-adjusted odds ratio = 5.86, 95% confidence interval, 1.10-31.16). CYP3A4*1B was associated inversely with the probability of having prostate cancer in Caucasians (age-adjusted odds ratio = 0.54, 95% confidence interval, 0.32 0.94). We also observed significant interactions among these loci associated with prostate cancer occurrence and severity. There were statistically significant differences in haplotype frequencies involving these three genes in high-stage cases (P < 0.05) compared with controls. The observation that CYP3A4 and CYP3A43 were associated with prostate cancer, are not in linkage equilibrium, and are both involved in testosterone metabolism, suggest that both CYP3A4*1B and CYP3A43*3 may influence the probability of having prostate cancer and disease severity. PMID- 15548720 TI - Reduced immunoglobulin E and allergy among adults with glioma compared with controls. AB - We and others have reported previously that adults with glioma are 1.5- to 4-fold less likely than controls to report a variety of allergic conditions. The consistent nature of this relationship calls for a biological explanation so that preventative or therapeutic modalities can be explored. We enrolled 403 newly diagnosed adult glioma cases in the San Francisco Bay Area over a 3-year period using a population-based cancer registry and 402 age/gender/ethnicity frequency matched controls identified via random digit dialing. We assessed total, food specific, and respiratory-specific IgE in available case (n = 228) and control (n = 289) serum samples. IgE levels were associated with gender, age, smoking status, and ethnicity among cases and/or controls. Among the cases, IgE levels were not associated with aspects of glioma therapy including radiation, chemotherapy, or tumor resection. Total IgE levels were lower in cases than controls: age/gender/ethnicity/education/smoking-adjusted odds ratio (OR) for elevated versus normal total IgE was 0.37 [95% confidence interval (CI), 0.22 0.64]. For the food panel, OR was 0.12 (95% CI, 0.04-0.41). For the respiratory panel, OR was 0.76 (95% CI, 0.52-1.1). Among respiratory allergies, late age of onset (>12 years) but not IgE levels defined a group with strong associations with risk (OR, 0.50; 95% CI, 0.33-0.75). These results corroborate and strengthen our findings of an inverse association between allergic reactions and glioma by showing a relationship with a biomarker for allergy and cancer for the first time. Furthermore, the results indicate a complex relationship between allergic disease and glioma risk that varies by allergen and allergic pathology. PMID- 15548721 TI - Sixteenth Annual Pezcoller Symposium: stem cells and epigenesis in cancer. AB - In the postgenomic era of cancer biology, it is becoming increasingly evident that epigenetic controls of gene expression play an important role in determining the phenotype of cancer cells, also indicating the possibility of restoring epigenetically a normal phenotype in cancer cells. Related to these mechanisms of control and their dynamic changes during carcinogenesis and tumor progression are the phenomena determining the relationships between stem cells and cancer cells. This symposium focused on epigenetic mechanisms affecting cancer development and possibly providing a basis for intervention. The basic biology of stem cells and the relationships between stem cells and cancer were discussed. Epigenetic control mechanisms affecting gene expression with emphasis on DNA methylation and histone function were considered. Therapeutic strategies stemming from the knowledge acquired in the basic topics discussed also were outlined. PMID- 15548722 TI - In vivo models for experimental therapeutics relevant to human cancer. PMID- 15548723 TI - Kallikrein 4 is a secreted protein. PMID- 15548724 TI - Combined angiotensin receptor blocker and ACE inhibitor on myocardial fibrosis and left ventricular stiffness in dogs with heart failure. AB - Angiotensin receptor blocker (ARB) and angiotensin-converting enzyme (ACE) inhibitor (ACEI) each act in a different manner to prevent myocardial fibrosis and left ventricular (LV) stiffness in animals with pathways in the heart for generating ANG II as well as ACE. A model of pacing-induced congestive heart failure (CHF) was used to test the central hypothesis that ARB + ACEI prevents myocardial fibrosis and decreases LV stiffness to a greater extent than ARB or ACEI alone. Thirty-five dogs were assigned to the following treatment protocols on the 8th day of a 4-wk pacing schedule: 1) rapid ventricular pacing, 2) ARB (candesartan cilexetil, 1.5 mg.kg(-1).day(-1)) with pacing, 3) ACEI (enalapril, 1.9 mg.kg(-1).day(-1)) with pacing, 4) ARB (candesartan cilexetil, 0.75 mg.kg( 1).day(-1)) + ACEI (enalapril, 0.95 mg.kg(-1).day(-1)) with pacing, and 5) sham operation. The LV stiffness coefficient was significantly increased after rapid pacing but was significantly lower with ARB + ACEI than with ARB or ACEI alone. The collagen volume fraction and mRNA levels of collagen I and III, which were increased by rapid pacing, were significantly lower with ARB + ACEI than with ARB or ACEI alone. Thus ARB + ACEI prevents myocardial fibrosis and decreases LV stiffness during the progression of CHF compared with ARB or ACEI alone. PMID- 15548725 TI - Bifurcation asymmetry of the porcine coronary vasculature and its implications on coronary flow heterogeneity. AB - The branching pattern of the coronary arteries and veins is asymmetric, i.e., many small vessels branch off of a large trunk such that the two daughter vessels at a bifurcation are of unequal diameters and lengths. One important implication of the geometric vascular asymmetry is the dispersion of blood flow at a bifurcation, which leads to large spatial heterogeneity of myocardial blood flow. To document the asymmetric branching pattern of the coronary vessels, we computed an asymmetry ratio for the diameters and lengths of all vessels, defined as the ratio of the daughter diameters and lengths, respectively. Previous data from silicone elastomer cast of the entire coronary vasculature including arteries, arterioles, venules, and veins were analyzed. Data on smaller vessels were obtained from histological specimens by optical sectioning, whereas data on larger vessels were obtained from vascular casts. Asymmetry ratios for vascular areas, volumes, resistances, and flows of the various daughter vessels were computed from the asymmetry ratios of diameters and lengths for every order of mother vessel. The results show that the largest orders of arterial and venous vessels are most asymmetric and the degree of asymmetry decreases toward the smaller vessels. Furthermore, the diameter asymmetry at a bifurcation is significantly larger for the coronary veins (1.7-6.8 for sinus veins) than the corresponding arteries (1.5-5.8 for left anterior descending coronary artery) for orders 2-10, respectively. The reported diameter asymmetry at a bifurcation leads to significant heterogeneity of blood flow at a bifurcation. Hence, the present data quantify the dispersion of blood flow at a bifurcation and are essential for understanding flow heterogeneity in the coronary circulation. PMID- 15548726 TI - Titin isoform-dependent effect of calcium on passive myocardial tension. AB - We studied the effects of Ca2+ on titin (connectin)-based passive tension in skinned myocardium expressing either predominantly N2B titin (rat right ventricle, RRV) or predominantly N2BA titin (bovine left atrium, BLA). Actomyosin based tension was abolished to undetectably low levels by selectively removing the thin filaments with a Ca2+-insensitive gelsolin fragment (FX-45). Myocardium was stretched in the presence and absence of Ca2+, and passive tension was measured. Ca2+ significantly increased passive tension during and after stretch in the BLA. The increase was insensitive to the actomyosin inhibitor 2,3 butanedione 2-monoxime, supporting the conclusion that the effect is titin based. Passive tension did not respond to calcium in the RRV, indicating that passive tension developed by N2B titin is calcium insensitive. Western blot analysis and immunofluorescence studies indicated that N2BA titin expresses E-rich PEVK motifs, whereas they are absent from N2B titin, supporting earlier single molecule studies that reported that E-rich motifs are required for calcium sensitivity. We conclude that calcium affects passive myocardial tension in a titin isoform-dependent manner. PMID- 15548727 TI - Stretch induces upregulation of key tyrosine kinase receptors in microvascular endothelial cells. AB - We previously demonstrated that cyclic stretch of cardiac myocytes activates paracrine signaling via vascular endothelial growth factor (VEGF) leading to angiogenesis. The present study tested the hypothesis that cyclic stretch upregulates tyrosine kinase receptors in rat coronary microvascular endothelial cells (RCMEC) and human umbilical vein endothelial cells (HUVEC). VEGF receptor-2 (Flk-1) protein levels increased in HUVEC and RCMEC in a time-dependent manner, but the increase occurred much earlier in RCMEC than in HUVEC. The enhancement of Flk-1 protein level was not inhibited by addition of VEGF neutralizing antibodies, indicating that VEGF is not involved in stretch-induced Flk-1 expression. VEGF receptor-1 (Flt-1) protein and mRNA were not changed by stretch. However, Tie-2 and Tie-1 protein levels increased in RCMEC. Angiopoietin-1 and 2, the ligands for Tie-2, increased in cardiac myocytes subjected to cyclic stretch but were not affected by stretch in endothelial cells (EC). Stretch or incubation of RCMEC with VEGF increased cell proliferation moderately, whereas stretch + VEGF had an additive effect on proliferation. Mechanical stretch induces upregulation of the key tyrosine kinase receptors Flk-1, Tie-2, and Tie-1 in vascular EC, which underlies the increase in sensitivity of EC to growth factors and, therefore, facilitates angiogenesis. These in vitro findings support the concept that stretch of cardiac myocytes and EC plays a key role in coronary angiogenesis. PMID- 15548728 TI - Responses of cerebral arterioles to ADP: eNOS-dependent and eNOS-independent mechanisms. AB - ADP mediates platelet-induced relaxation of blood vessels and may function as an important intercellular signaling molecule in the brain. We used pharmacological and genetic approaches to examine mechanisms that mediate responses of cerebral arterioles to ADP, including the role of endothelial nitric oxide synthase (eNOS). We examined responses of cerebral arterioles (control diameter approximately 30 microm) in anesthetized wild-type (WT, eNOS+/+) and eNOS deficient (eNOS-/-) mice using a cranial window. In WT mice, local application of ADP produced vasodilation that was not altered by indomethacin but was reduced by approximately 50% by NG-nitro-L-arginine (L-NNA) or 1H-[1,2,4]oxadiazolo[4,3 a]quinoxalin-1-one (ODQ) (inhibitors of NOS and soluble guanylate cyclase, respectively). In eNOS-/- mice, responses to ADP were largely preserved, and a significant component of the response was resistant to L-NNA (a finding similar to that in WT mice treated with L-NNA). In the absence of L-NNA, responses to ADP were markedly reduced by charybdotoxin plus apamin [inhibitors of Ca2+-dependent K+ channels and responses mediated by endothelium-derived hyperpolarizing factor (EDHF)] in both WT and eNOS-/- mice. Thus pharmacological and genetic evidence suggests that a significant portion of the response to ADP in cerebral microvessels is mediated by a mechanism independent of eNOS. The eNOS-independent mechanism is functional in the absence of inhibited eNOS and most likely is mediated by an EDHF. PMID- 15548729 TI - A self-calibrating telemetry system for measurement of ventricular pressure volume relations in conscious, freely moving rats. AB - Using Bluetooth wireless technology, we developed an implantable telemetry system for measurement of the left ventricular pressure-volume relation in conscious, freely moving rats. The telemetry system consisted of a pressure-conductance catheter (1.8-Fr) connected to a small (14-g) fully implantable signal transmitter. To make the system fully telemetric, calibrations such as blood resistivity and parallel conductance were also conducted telemetrically. To estimate blood resistivity, we used four electrodes arranged 0.2 mm apart on the pressure-conductance catheter. To estimate parallel conductance, we used a dual frequency method. We examined the accuracy of calibrations, stroke volume (SV) measurements, and the reproducibility of the telemetry. The blood resistivity estimated telemetrically agreed with that measured using an ex vivo cuvette method (y=1.09x - 11.9, r2= 0.88, n=10). Parallel conductance estimated by the dual-frequency (2 and 20 kHz) method correlated well with that measured by a conventional saline injection method (y=1.59x - 1.77, r2= 0.87, n=13). The telemetric SV closely correlated with the flowmetric SV during inferior vena cava occlusions (y=0.96x + 7.5, r2=0.96, n=4). In six conscious rats, differences between the repeated telemetries on different days (3 days apart on average) were reasonably small: 13% for end-diastolic volume, 20% for end-systolic volume, 28% for end-diastolic pressure, and 6% for end-systolic pressure. We conclude that the developed telemetry system enables us to estimate the pressure-volume relation with reasonable accuracy and reproducibility in conscious, untethered rats. PMID- 15548730 TI - An overview of cell renewal in the testis throughout the reproductive cycle of a seasonal breeding teleost, the gilthead seabream (Sparus aurata L). AB - The gilthead seabream is a protandrous hermaphrodite seasonal breeding teleost with a bisexual gonad that offers an interesting model for studying the testicular regression process that occurs in both seasonal testicular involution and sex change. Insofar as fish reproduction is concerned, little is known about cell renewal and elimination during the reproductive cycle of seasonal breeding teleosts with asynchronous spermatogenesis. We have previously described how acidophilic granulocytes infiltrate the testis during postspawning where, surprisingly, they produce interleukin-1beta, a known growth factor for mammalian spermatogonia, rather than being directly involved in the elimination of degenerative germ cells. In this study, we are able to discriminate between spermatogonia stem cells and primary spermatogonia according to their nuclear and cytoplasmic diameters and location in the germinal epithelium, finding that these two cell types, together with Sertoli cells, proliferate throughout the reproductive cycle with a rate that depends on the reproductive stage. Thus, during spermatogenesis the spermatogonia stem cells, the Sertoli cells, and the developing germ cells (primary spermatogonia, A and B spermatogonia, and spermatocytes) in the germinal compartment, and cells with fibroblast-shaped nuclei in the interstitial tissue proliferate. However, during spawning, the testis shows few proliferating cells. During postspawning, the resumption of proliferation, the occurrence of apoptotic spermatogonia, and the phagocytosis of nonshed spermatozoa by Sertoli cells lead to a reorganization of both the germinal compartment and the interstitial tissue. Finally, the proliferation of spermatogonia increases during resting when, unexpectedly, both oogonia and oocytes also proliferate. This proliferative pattern was correlated with the gonadosomatic index, testicular morphology, and testicular and gonad areas, suggesting that complex mechanisms operate in the regulation of gonocyte proliferation in hermaphrodite fish. PMID- 15548731 TI - Infertility in 5,10-methylenetetrahydrofolate reductase (MTHFR)-deficient male mice is partially alleviated by lifetime dietary betaine supplementation. AB - Metabolism of folate is essential for proper cellular function. Within the folate pathway, methylenetetrahydrofolate reductase (MTHFR) reduces 5,10 methylenetetrahydrofolate to 5-methyltetrahydrofolate, a methyl donor for remethylation of homocysteine to methionine, the precursor of S adenosylmethionine. S-adenosylmethionine is the methyl donor for numerous cellular reactions. In adult male mice, MTHFR levels are highest in the testis; this finding, in conjunction with recent clinical evidence, suggest an important role for MTHFR in spermatogenesis. Indeed, we show here that severe MTHFR deficiency in male mice results in abnormal spermatogenesis and infertility. Maternal oral administration of betaine, an alternative methyl donor, throughout pregnancy and nursing, resulted in improved testicular histology in Mthfr-/- offspring at Postnatal Day 6, but not at 8 mo of age. However, when betaine supplementation was maintained postweaning, testicular histology improved, and sperm numbers and fertility increased significantly. We postulate that the adverse effects of MTHFR deficiency on spermatogenesis, may, in part, be mediated by alterations in the transmethylation pathway and suggest that betaine supplementation may provide a means to bypass MTHFR deficiency and its adverse effects on spermatogenesis by maintaining normal methylation levels within male germ cells. PMID- 15548732 TI - Term human fetal membranes have a weak zone overlying the lower uterine pole and cervix before onset of labor. AB - The etiology of fetal membrane (FM) rupture is unknown. A hypothesis that the FM weakens by a process of collagen remodeling and apoptosis to facilitate rupture has been proposed. Human FMs reportedly exhibit a zone of altered histology, postulated to be the FM rupture site, but concomitant FM weakness has not been demonstrated. We hypothesized that a discrete zone of FM with marked weakness, histological change, and evidence of remodeling and apoptosis, develops in late gestation in the FM overlying the cervix. FM tissue from women undergoing prelabor cesarean delivery were perioperatively marked to identify the FM overlying the cervix, cut with a procedure that facilitates remapping the rupture strength of FM pieces to their former location and orientation on a three dimensional model, and tested for strength. A 10-cm FM zone centered at the cervical mark was compared with the remaining FM. Mean rupture strength within the cervical zone was 55% of the remaining FM. The cervical zone also exhibited increased MMP-9 protein, decreased tissue inhibitor of metalloproteinases-3 (TIMP 3) protein, and increased PARP cleavage coincident with the previously reported zone of altered histology. A discrete zone of weakness is present in term prelabor FMs overlying the cervix and has biochemical characteristics consistent with tissue remodeling and apoptosis. PMID- 15548733 TI - Effects of nicotine on human luteal cells in vitro: a possible role on reproductive outcome for smoking women. AB - We investigated the effect of nicotine and its methylated metabolite, N-methyl nicotine (M-nicotine), on human luteal cells by measuring release of progesterone and prostaglandins (PGs) from cultured cells and by testing gene expression of vascular endothelial growth factor (VEGF), an angiogenic factor strictly involved in luteal pathophysiology. Primary cultures of human luteal cells were treated for 24 h with nicotine and M-nicotine (from 10(-6) to 10(-11) M) either alone or combined with hCG (25 ng/ml); progesterone and PGs were assayed in the culture medium. In another group of experiments, luteal cells were treated for 24 h with nicotine and M-nicotine (10(-7) M) to perform reverse transcriptase-polymerase chain reaction on VEGF mRNA. Nicotine and M-nicotine negatively affected basal luteal steroidogenesis at all tested concentrations, but neither was able to affect hCG-induced progesterone release. Both substances were able to significantly increase PGF2alpha release from luteal cells, with a dose-related efficacy for M-nicotine. On the contrary, PGE2 release was significantly inhibited by both nicotine and its metabolite. Finally, nicotine was able to increase VEGF mRNA expression significantly, whereas M-nicotine was not. In conclusion, nicotine and M-nicotine can induce a sort of luteal insufficiency by inhibiting progesterone release, probably through modulation of the PG system. PMID- 15548734 TI - Diversification and alteration of recognition specificity of the pollen ligand SP11/SCR in self-incompatibility of Brassica and Raphanus. AB - The recognition specificity of the pollen ligand of self-incompatibility (SP11/SCR) was investigated using Brassica rapa transgenic plants expressing SP11 transgenes, and SP11 of Raphanus sativus S-21 was found to have the same recognition specificity as that of B. rapa S-9. In a set of three S haplotypes, whose sequence identities of SP11 and SRK are fairly high, R. sativus S-6 showed the same recognition specificity as Brassica oleracea S-18 and a slightly different specificity from B. rapa S-52. B. oleracea S-18, however, showed a different specificity from B. rapa S-52. Using these similar S haplotypes, chimeric SP11 proteins were produced by domain swapping. Bioassay using the chimeric SP11 proteins revealed that the incompatibility response induction activity was altered by the replacement of Region III and Region V. Pollen grains of Brassica transgenic plants expressing chimeric SP11 of the B. oleracea SP11-18 sequence with Region III and Region V from B. rapa SP11-52 (chimeric BoSP11 18[52]) were partially incompatible with the B. rapa S-52 stigmas, and those expressing the R. sativus SP11-6 sequence with Region III and Region V from B. rapa SP11-52 (chimeric RsSP11-6[52]) were completely incompatible with the stigmas having B. rapa S-52. However, the transgenic plant expressing chimeric RsSP11-6(52) also showed incompatibility with B. oleracea S-18 stigmas. These results suggest that Regions III and Region V of SP11 are important for determining the recognition specificity, but not the sole determinant. A possible process of the generation of a new S haplotype is herein discussed. PMID- 15548735 TI - Arabidopsis WPP-domain proteins are developmentally associated with the nuclear envelope and promote cell division. AB - The nuclear envelope (NE) acts as a selective barrier to macromolecule trafficking between the nucleus and the cytoplasm and undergoes a complex reorganization during mitosis. Different eukaryotic kingdoms show specializations in NE function and composition. In contrast with vertebrates, the protein composition of the NE and the function of NE proteins are barely understood in plants. MFP1 attachment factor 1 (MAF1) is a plant-specific NE-associated protein first identified in tomato (Lycopersicon esculentum). Here, we demonstrate that two Arabidopsis thaliana MAF1 homologs, WPP1 and WPP2, are associated with the NE specifically in undifferentiated cells of the root tip. Reentry into cell cycle after callus induction from differentiated root segments reprograms their NE association. Based on green fluorescent protein fusions and immunogold labeling data, the proteins are associated with the outer NE and the nuclear pores in interphase cells and with the immature cell plate during cytokinesis. RNA interference-based suppression of the Arabidopsis WPP family causes shorter primary roots, a reduced number of lateral roots, and reduced mitotic activity of the root meristem. Together, these data demonstrate the existence of regulated NE targeting in plants and identify a class of plant-specific NE proteins involved in mitotic activity. PMID- 15548736 TI - Chloroplast elongation factor ts pro-protein is an evolutionarily conserved fusion with the s1 domain-containing plastid-specific ribosomal protein-7. AB - The components of chloroplast translation are similar to those of prokaryotic translation but contain some additional unique features. Proteomic analysis of the Chlamydomonas reinhardtii chloroplast ribosome identified an S1-like protein, plastid-specific ribosomal protein-7 (PSRP-7), as a stoichiometric component of the 30S subunit. Here, we report that PSRP-7 is part of a polyprotein that contains PSRP-7 on its amino end and two translation elongation factor Ts (EF-Ts) domains at the carboxy end. We named this polyprotein PETs (for polyprotein of EF Ts). Pets is a single-copy gene containing the only chloroplast PSRP-7 and EF-Ts sequences found in the C. reinhardtii genome. The pets precursor transcript undergoes alternative splicing to generate three mRNAs with open reading frames (ORFs) of 1.68, 1.8, and 3 kb. A 110-kD pro-protein is translated from the 3-kb ORF, and the majority of this protein is likely posttranslationally processed into the 65-kD protein PSRP-7 and a 55-kD EF-Ts. PETs homologs are found in Arabidopsis thaliana and rice (Oryza sativa). The conservation of the 110-kD PETs polyprotein in the plant kingdom suggests that PSRP-7 and EF-Ts function together in some aspects of chloroplast translation and that the PETs pro-protein may have a novel function as a whole. PMID- 15548737 TI - Solution structure of the B3 DNA binding domain of the Arabidopsis cold responsive transcription factor RAV1. AB - The B3 DNA binding domain is shared amongst various plant-specific transcription factors, including factors involved in auxin-regulated and abscisic acid regulated transcription. Herein, we report the NMR solution structure of the B3 domain of the Arabidopsis thaliana cold-responsive transcription factor RAV1. The structure consists of a seven-stranded open beta-barrel and two alpha-helices located at the ends of the barrel and is significantly similar to the structure of the noncatalytic DNA binding domain of the restriction enzyme EcoRII. An NMR titration experiment revealed a DNA recognition interface that enabled us to propose a structural model of the protein-DNA complex. The locations of the DNA contacting residues are also likely to be similar to those of the EcoRII DNA binding domain. PMID- 15548738 TI - A Robot-based platform to measure multiple enzyme activities in Arabidopsis using a set of cycling assays: comparison of changes of enzyme activities and transcript levels during diurnal cycles and in prolonged darkness. AB - A platform has been developed to measure the activity of 23 enzymes that are involved in central carbon and nitrogen metabolism in Arabidopsis thaliana. Activities are assayed in optimized stopped assays and the product then determined using a suite of enzyme cycling assays. The platform requires inexpensive equipment, is organized in a modular manner to optimize logistics, calculates results automatically, combines high sensitivity with throughput, can be robotized, and has a throughput of three to four activities in 100 samples per person/day. Several of the assays, including those for sucrose phosphate synthase, ADP glucose pyrophosphorylase (AGPase), ferredoxin-dependent glutamate synthase, glycerokinase, and shikimate dehydrogenase, provide large advantages over previous approaches. This platform was used to analyze the diurnal changes of enzyme activities in wild-type Columbia-0 (Col-0) and the starchless plastid phosphoglucomutase (pgm) mutant, and in Col-0 during a prolongation of the night. The changes of enzyme activities were compared with the changes of transcript levels determined with the Affymetrix ATH1 array. Changes of transcript levels typically led to strongly damped changes of enzyme activity. There was no relation between the amplitudes of the diurnal changes of transcript and enzyme activity. The largest diurnal changes in activity were found for AGPase and nitrate reductase. Examination of the data and comparison with the literature indicated that these are mainly because of posttranslational regulation. The changes of enzyme activity are also strongly delayed, with the delay varying from enzyme to enzyme. It is proposed that enzyme activities provide a quasi-stable integration of regulation at several levels and provide useful data for the characterization and diagnosis of different physiological states. As an illustration, a decision tree constructed using data from Col-0 during diurnal changes and a prolonged dark treatment was used to show that, irrespective of the time of harvest during the diurnal cycle, the pgm mutant resembles a wild-type plant that has been exposed to a 3 d prolongation of the night. PMID- 15548740 TI - The N terminus of bacterial elongation factor Tu elicits innate immunity in Arabidopsis plants. AB - Innate immunity is based on the recognition of pathogen-associated molecular patterns (PAMPs). Here, we show that elongation factor Tu (EF-Tu), the most abundant bacterial protein, acts as a PAMP in Arabidopsis thaliana and other Brassicaceae. EF-Tu is highly conserved in all bacteria and is known to be N acetylated in Escherichia coli. Arabidopsis plants specifically recognize the N terminus of the protein, and an N-acetylated peptide comprising the first 18 amino acids, termed elf18, is fully active as inducer of defense responses. The shorter peptide, elf12, comprising the acetyl group and the first 12 N-terminal amino acids, is inactive as elicitor but acts as a specific antagonist for EF-Tu related elicitors. In leaves of Arabidopsis plants, elf18 induces an oxidative burst and biosynthesis of ethylene, and it triggers resistance to subsequent infection with pathogenic bacteria. PMID- 15548739 TI - Translational regulation via 5' mRNA leader sequences revealed by mutational analysis of the Arabidopsis translation initiation factor subunit eIF3h. AB - Eukaryotic translation initiation factor 3 (eIF3) consists of core subunits that are conserved from yeast to man as well as less conserved, noncore, subunits with potential regulatory roles. Whereas core subunits tend to be indispensable for cell growth, the roles of the noncore subunits remain poorly understood. We addressed the hypothesis that eIF3 noncore subunits have accessory functions that help to regulate translation initiation, by focusing on the Arabidopsis thaliana eIF3h subunit. Indeed, eIF3h was not essential for general protein translation. However, results from transient expression assays and polysome fractionation indicated that the translation efficiency of specific 5' mRNA leader sequences was compromised in an eif3h mutant, including the mRNA for the basic domain leucine zipper (bZip) transcription factor ATB2/AtbZip11, translation of which is regulated by sucrose. Among other pleiotropic developmental defects, the eif3h mutant required exogenous sugar to transit from seedling to vegetative development, but it was hypersensitive to elevated levels of exogenous sugars. The ATB2 mRNA was rendered sensitive to the eIF3h level by a series of upstream open reading frames. Moreover, eIF3h could physically interact with subunits of the COP9 signalosome, a protein complex implicated primarily in the regulation of protein ubiquitination, supporting a direct biochemical connection between translation initiation and protein turnover. Together, these data implicate eIF3 in mRNA-associated translation initiation events, such as scanning, start codon recognition, or reinitiation and suggest that poor translation initiation of specific mRNAs contributes to the pleiotropic spectrum of phenotypic defects in the eif3h mutant. PMID- 15548741 TI - RAR1 positively controls steady state levels of barley MLA resistance proteins and enables sufficient MLA6 accumulation for effective resistance. AB - The polymorphic barley (Hordeum vulgare) Mla locus harbors allelic race-specific resistance (R) genes to the powdery mildew fungus Blumeria graminis f sp hordei. The highly sequence-related MLA proteins contain an N-terminal coiled-coil structure, a central nucleotide binding (NB) site, a Leu-rich repeat (LRR) region, and a C-terminal non-LRR region. Using transgenic barley lines expressing epitope-tagged MLA1 and MLA6 derivatives driven by native regulatory sequences, we show a reversible and salt concentration-dependent distribution of the intracellular MLA proteins in soluble and membrane-associated pools. A posttranscriptional process directs fourfold greater accumulation of MLA1 over MLA6. Unexpectedly, in rar1 mutant plants that are compromised for MLA6 but not MLA1 resistance, the steady state level of both MLA isoforms is reduced. Furthermore, differential steady state levels of MLA1/MLA6 hybrid proteins correlate with their requirement for RAR1; the RAR1-independent hybrid protein accumulates to higher levels and the RAR1-dependent one to lower levels. Interestingly, yeast two-hybrid studies reveal that the LRR domains of RAR1 independent but not RAR1-dependent MLA isoforms interact with SGT1, a RAR1 interacting protein required for the function of many NB-LRR type R proteins. Our findings implicate the existence of a conserved mechanism to reach minimal NB-LRR R protein thresholds that are needed to trigger effective resistance responses. PMID- 15548742 TI - Expression and functional roles of the two distinct NDH-1 complexes and the carbon acquisition complex NdhD3/NdhF3/CupA/Sll1735 in Synechocystis sp PCC 6803. AB - To investigate the (co)expression, interaction, and membrane location of multifunctional NAD(P)H dehydrogenase type 1 (NDH-1) complexes and their involvement in carbon acquisition, cyclic photosystem I, and respiration, we grew the wild type and specific ndh gene knockout mutants of Synechocystis sp PCC 6803 under different CO2 and pH conditions, followed by a proteome analysis of their membrane protein complexes. Typical NDH-1 complexes were represented by NDH-1L (large) and NDH-1M (medium size), located in the thylakoid membrane. The NDH-1L complex, missing from the DeltaNdhD1/D2 mutant, was a prerequisite for photoheterotrophic growth and thus apparently involved in cellular respiration. The amount of NDH-1M and the rate of P700+ rereduction in darkness in the DeltaNdhD1/D2 mutant grown at low CO2 were similar to those in the wild type, whereas in the M55 mutant (DeltaNdhB), lacking both NDH-1L and NDH-1M, the rate of P700+ rereduction was very slow. The NDH-1S (small) complex, localized to the thylakoid membrane and composed of only NdhD3, NdhF3, CupA, and Sll1735, was strongly induced at low CO2 in the wild type as well as in DeltaNdhD1/D2 and M55. In contrast with the wild type and DeltaNdhD1/D2, which show normal CO2 uptake, M55 is unable to take up CO2 even when the NDH-1S complex is present. Conversely, the DeltaNdhD3/D4 mutant, also unable to take up CO2, lacked NDH-1S but exhibited wild-type levels of NDH-1M at low CO2. These results demonstrate that both NDH-1S and NDH-1M are essential for CO2 uptake and that NDH-1M is a functional complex. We also show that the Na+/HCO3- transporter (SbtA complex) is located in the plasma membrane and is strongly induced in the wild type and mutants at low CO2. PMID- 15548743 TI - Antagonistic interaction between abscisic acid and jasmonate-ethylene signaling pathways modulates defense gene expression and disease resistance in Arabidopsis. AB - The plant hormones abscisic acid (ABA), jasmonic acid (JA), and ethylene are involved in diverse plant processes, including the regulation of gene expression during adaptive responses to abiotic and biotic stresses. Previously, ABA has been implicated in enhancing disease susceptibility in various plant species, but currently very little is known about the molecular mechanisms underlying this phenomenon. In this study, we obtained evidence that a complex interplay between ABA and JA-ethylene signaling pathways regulate plant defense gene expression and disease resistance. First, we showed that exogenous ABA suppressed both basal and JA-ethylene-activated transcription from defense genes. By contrast, ABA deficiency as conditioned by the mutations in the ABA1 and ABA2 genes, which encode enzymes involved in ABA biosynthesis, resulted in upregulation of basal and induced transcription from JA-ethylene responsive defense genes. Second, we found that disruption of AtMYC2 (allelic to JASMONATE INSENSITIVE1 [JIN1]), encoding a basic helix-loop-helix Leu zipper transcription factor, which is a positive regulator of ABA signaling, results in elevated levels of basal and activated transcription from JA-ethylene responsive defense genes. Furthermore, the jin1/myc2 and aba2-1 mutants showed increased resistance to the necrotrophic fungal pathogen Fusarium oxysporum. Finally, using ethylene and ABA signaling mutants, we showed that interaction between ABA and ethylene signaling is mutually antagonistic in vegetative tissues. Collectively, our results indicate that the antagonistic interactions between multiple components of ABA and the JA ethylene signaling pathways modulate defense and stress responsive gene expression in response to biotic and abiotic stresses. PMID- 15548745 TI - Structural basis for the entrance into the phenylpropanoid metabolism catalyzed by phenylalanine ammonia-lyase. AB - Because of its key role in secondary phenylpropanoid metabolism, Phe ammonia lyase is one of the most extensively studied plant enzymes. To provide a basis for detailed structure-function studies, the enzyme from parsley (Petroselinum crispum) was crystallized, and the structure was elucidated at 1.7-A resolution. It contains the unusual electrophilic 4-methylidene-imidazole-5-one group, which is derived from a tripeptide segment in two autocatalytic dehydration reactions. The enzyme resembles His ammonia-lyase from the general His degradation pathway but contains 207 additional residues, mainly in an N-terminal extension rigidifying a domain interface and in an inserted alpha-helical domain restricting the access to the active center. Presumably, Phe ammonia-lyase developed from His ammonia-lyase when fungi and plants diverged from the other kingdoms. A pathway of the catalyzed reaction is proposed in agreement with established biochemical data. The inactivation of the enzyme by a nucleophile is described in detail. PMID- 15548744 TI - Heterodimerization and endocytosis of Arabidopsis brassinosteroid receptors BRI1 and AtSERK3 (BAK1). AB - In Arabidopsis thaliana brassinosteroid (BR), perception is mediated by two Leu rich repeat receptor-like kinases, BRASSINOSTEROID INSENSITIVE1 (BRI1) and BRI1 ASSOCIATED RECEPTOR KINASE1 (BAK1) (Arabidopsis SOMATIC EMBRYOGENESIS RECEPTOR like KINASE3 [AtSERK3]). Genetic, biochemical, and yeast (Saccharomyces cerevisiae) interaction studies suggested that the BRI1-BAK1 receptor complex initiates BR signaling, but the role of the BAK1 receptor is still not clear. Using transient expression in protoplasts of BRI1 and AtSERK3 fused to cyan and yellow fluorescent green fluorescent protein variants allowed us to localize each receptor independently in vivo. We show that BRI1, but not AtSERK3, homodimerizes in the plasma membrane, whereas BRI1 and AtSERK3 preferentially heterodimerize in the endosomes. Coexpression of BRI1 and AtSERK3 results in a change of the steady state distribution of both receptors because of accelerated endocytosis. Endocytic vesicles contain either BRI1 or AtSERK3 alone or both. We propose that the AtSERK3 protein is involved in changing the equilibrium between plasma membrane-located BRI1 homodimers and endocytosed BRI1-AtSERK3 heterodimers. PMID- 15548746 TI - The role of [Delta]1-pyrroline-5-carboxylate dehydrogenase in proline degradation. AB - In response to stress, plants accumulate Pro, requiring degradation after release from adverse conditions. Delta1-Pyrroline-5-carboxylate dehydrogenase (P5CDH), the second enzyme for Pro degradation, is encoded by a single gene expressed ubiquitously. To study the physiological function of P5CDH, T-DNA insertion mutants in AtP5CDH were isolated and characterized. Although Pro degradation was undetectable in p5cdh mutants, neither increased Pro levels nor an altered growth phenotype were observed under normal conditions. Thus AtP5CDH is essential for Pro degradation but not required for vegetative plant growth. External Pro application caused programmed cell death, with callose deposition, reactive oxygen species production, and DNA laddering, involving a salicylic acid signal transduction pathway. p5cdh mutants were hypersensitive toward Pro and other molecules producing P5C, such as Arg and Orn. Pro levels were the same in the wild type and mutants, but P5C was detectable only in p5cdh mutants, indicating that P5C accumulation may be the cause for Pro hypersensitivity. Accordingly, overexpression of AtP5CDH resulted in decreased sensitivity to externally supplied Pro. Thus, Pro and P5C/Glu semialdehyde may serve as a link between stress responses and cell death. PMID- 15548748 TI - Isolation and characterization of active LINE and SINEs from the eel. AB - Long interspersed elements (LINEs) and short interspersed elements (SINEs) are retrotransposons. These elements can mobilize by the "copy-and-paste" mechanism, in which their own RNA is reverse-transcribed into complementary DNA (cDNA). LINEs and SINEs not only are components of eukaryotic genomes but also drivers of genomic evolution. Thus, studies of the amplification mechanism of LINEs and SINEs are important for understanding eukaryotic genome evolution. Here we report the characterization of one LINE family (UnaL2) and two SINE families (UnaSINE1 and UnaSINE2) from the eel (Anguilla japonica) genome. UnaL2 is approximately 3.6 kilobases (kb) and encodes only one open reading frame (ORF). UnaL2 belongs to the stringent type--thought to be a major group of LINEs--and can mobilize in HeLa cells. We also show that UnaL2 and the two UnaSINEs have similar 3' tails, and that both UnaSINE1 and UnaSINE2 can be mobilized by UnaL2 in HeLa cells. These elements are thus useful for delineating the amplification mechanism of stringent type LINEs as well as that of SINEs. PMID- 15548747 TI - Genetic structure of Hmong-Mien speaking populations in East Asia as revealed by mtDNA lineages. AB - Hmong-Mien (H-M) is a major language family in East Asia, and its speakers distribute primarily in southern China and Southeast Asia. To date, genetic studies on H-M speaking populations are virtually absent in the literature. In this report, we present the results of an analysis of genetic variations in the mitochondrial DNA (mtDNA) hypervariable segment 1 (HVS1) region and diagnostic variants in the coding regions in 537 individuals sampled from 17 H-M populations across East Asia. The analysis showed that the haplogroups that are predominant in southern East Asia, including B, R9, N9a, and M7, account for 63% (ranging from 45% to 90%) of mtDNAs in H-M populations. Furthermore, analysis of molecular variance (AMOVA), phylogenetic tree analysis, and principal component (PC) analysis demonstrate closer relatedness between H-M and other southern East Asians, suggesting a general southern origin of maternal lineages in the H-M populations. The estimated ages of the mtDNA lineages that are specific to H-M coincide with those based on archeological cultures that have been associated with H-M. Analysis of genetic distance and phylogenetic tree indicated some extent of difference between the Hmong and the Mien populations. Together with the higher frequency of north-dominating lineages observed in the Hmong people, our results indicate that the Hmong populations had experienced more contact with the northern East Asians, a finding consistent with historical evidence. Moreover, our data defined some new (sub-)haplogroups (A6, B4e, B4f, C5, F1a1, F1a1a, and R9c), which will direct further efforts to improve the phylogeny of East Asian mtDNAs. PMID- 15548749 TI - Identification of a Giardia krr1 homolog gene and the secondarily anucleolate condition of Giaridia lamblia. AB - Giaridia lamblia was long considered to be one of the most primitive eukaryotes and to lie close to the transition between prokaryotes and eukaryotes, but several supporting features, such as lack of mitochondrion and Golgi, have been challenged recently. It was also reported previously that G. lamblia lacked nucleolus, which is the site of pre-rRNA processing and ribosomal assembling in the other eukaryotic cells. Here, we report the identification of the yeast homolog gene, krr1, in the anucleolate eukaryote, G. lamblia. The krr1 gene, encoding one of the pre-rRNA processing proteins in yeast, is actively transcribed in G. lamblia. The deduced protein sequence of G. lamblia krr1 is highly similar to yeast KRR1p that contains a single-KH domain. Our database searches indicated that krr1 genes actually present in diverse eukaryotes and also seem to present in Archaea. However, only the eukaryotic homologs, including that of G. lamblia, have the single-KH domain, which contains the conserved motif KR(K)R. Fibrillarin, another important pre-rRNA processing protein has also been identified previously in G. lamblia. Moreover, our database search shows that nearly half of the other nucleolus-localized protein genes of eukaryotic cells also have their homologs in Giardia. Therefore, we suggest that a common mechanism of pre-RNA processing may operate in the anucleolate eukaryote G. lamblia and in the other eukaryotes and that like the case of "lack of mitochondrion," "lack of nucleolus" may not be a primitive feature, but a secondarily evolutionary condition of the parasite. PMID- 15548750 TI - Multidimensional vector space representation for convergent evolution and molecular phylogeny. AB - With growing amounts of genome data and constant improvement of models of molecular evolution, phylogenetic reconstruction became more reliable. However, our knowledge of the real process of molecular evolution is still limited. When enough large-sized data sets are analyzed, any subtle biases in statistical models can support incorrect topologies significantly because of the high signal to-noise ratio. We propose a procedure to locate sequences in a multidimensional vector space (MVS), in which the geometry of the space is uniquely determined in such a way that the vectors of sequence evolution are orthogonal among different branches. In this paper, the MVS approach is developed to detect and remove biases in models of molecular evolution caused by unrecognized convergent evolution among lineages or unexpected patterns of substitutions. Biases in the estimated pairwise distances are identified as deviations (outliers) of sequence spatial vectors from the expected orthogonality. Modifications to the estimated distances are made by minimizing an index to quantify the deviations. In this way, it becomes possible to reconstruct the phylogenetic tree, taking account of possible biases in the model of molecular evolution. The efficacy of the modification procedure was verified by simulating evolution on various topologies with rate heterogeneity and convergent change. The phylogeny of placental mammals in previous analyses of large data sets has varied according to the genes being analyzed. Systematic deviations caused by convergent evolution were detected by our procedure in all representative data sets and were found to strongly affect the tree structure. However, the bias correction yielded a consistent topology among data sets. The existence of strong biases was validated by examining the sites of convergent evolution between the hedgehog and other species in mitochondrial data set. This convergent evolution explains why it has been difficult to determine the phylogenetic placement of the hedgehog in previous studies. PMID- 15548751 TI - Accounting for uncertainty in the tree topology has little effect on the decision theoretic approach to model selection in phylogeny estimation. AB - Currently available methods for model selection used in phylogenetic analysis are based on an initial fixed-tree topology. Once a model is picked based on this topology, a rigorous search of the tree space is run under that model to find the maximum-likelihood estimate of the tree (topology and branch lengths) and the maximum-likelihood estimates of the model parameters. In this paper, we propose two extensions to the decision-theoretic (DT) approach that relax the fixed topology restriction. We also relax the fixed-topology restriction for the Bayesian information criterion (BIC) and the Akaike information criterion (AIC) methods. We compare the performance of the different methods (the relaxed, restricted, and the likelihood-ratio test [LRT]) using simulated data. This comparison is done by evaluating the relative complexity of the models resulting from each method and by comparing the performance of the chosen models in estimating the true tree. We also compare the methods relative to one another by measuring the closeness of the estimated trees corresponding to the different chosen models under these methods. We show that varying the topology does not have a major impact on model choice. We also show that the outcome of the two proposed extensions is identical and is comparable to that of the BIC, Extended BIC, and DT. Hence, using the simpler methods in choosing a model for analyzing the data is more computationally feasible, with results comparable to the more computationally intensive methods. Another outcome of this study is that earlier conclusions about the DT approach are reinforced. That is, LRT, Extended-AIC, and AIC result in more complicated models that do not contribute to the performance of the phylogenetic inference, yet cause a significant increase in the time required for data analysis. PMID- 15548752 TI - Patterns of diversifying selection in the phytotoxin-like scr74 gene family of Phytophthora infestans. AB - Phytophthora infestans, the organism responsible for the Irish famine, causes late blight, a re-emerging disease of potato and tomato. Little is known about the molecular evolution of P. infestans genes. To identify candidate effector genes (virulence or avirulence genes) that may have co-evolved with the host, we mined expressed sequence tag (EST) data from infection stages of P. infestans for secreted and potentially polymorphic genes. This led to the identification of scr74, a gene that encodes a predicted 74-amino acid secreted cysteine-rich protein with similarity to the Phytophthora cactorum phytotoxin PcF. The expression of scr74 was upregulated approximately 60-fold 2 to 4 days after inoculation of tomato and was also significantly induced during early stages of colonization of potato. The scr74 gene was found to belong to a highly polymorphic gene family within P. infestans with 21 different sequences identified. Using the approximate and maximum likelihood (ML) methods, we found that diversifying selection likely caused the extensive polymorphism observed within the scr74 gene family. Pairwise comparisons of 17 scr74 sequences revealed elevated ratios of nonsynonymous to synonymous nucleotide-substitution rates, particularly in the mature region of the proteins. Using ML, all 21 polymorphic amino acid sites were identified to be under diversifying selection. Of these 21 amino acids, 19 are located in the mature protein region, suggesting that selection may have acted on the functional portions of the proteins. Further investigation of gene copy number and organization revealed that the scr74 gene family comprises at least three copies located in a region of no more than 300 kb of the P. infestans genome. We found evidence that recombination contributed to sequence divergence within at least one gene locus. These results led us to propose an evolutionary model that involves gene duplication and recombination, followed by functional divergence of scr74 genes. This study provides support for using diversifying selection as a criterion for identifying candidate effector genes from sequence databases. PMID- 15548753 TI - Prevention of mutagen formation in heated meats and model systems. AB - Possible means for preventing mutagen formation in cooked meats and in heated model systems are described. One way to reduce mutagenicity in cooked meats is to control cooking temperature, time and method. Another way is to increase water content or to avoid loss of water in meats during cooking. Addition of an excessive amount of reducing sugars to meats before cooking is effective in minimizing mutagen formation, which may be due to suppression of generation of the pyrazine cation radical Maillard intermediate of heterocyclic amines. Addition of a small amount of ascorbate or erythorbate is also effective, which may be the result of scavenging the intermediary pyrazine cation radical. PMID- 15548754 TI - Genotoxicity induced by saponified coconut oil surfactant in prokaryote systems. AB - Surfactants are amphiphilic substances with special properties and chemical structures that allow a reduction in interfacial tension, which permits an increase in molecule solubilization. The critical micelle concentration (CMC) is an important characteristic of surfactants that determines their aggregate state, which is generally related to its functional mechanism. In this work the genotoxic potential of saponified coconut oil (SCO), a surfactant obtained from Cocos nucifera, was analyzed using prokaryote systems. DNA strand breaks were not observed after treatment of a plasmid with SCO. Negative results were also obtained in the SOS Chromotest using Escherichia coli strains PQ35 and PQ37. A moderate toxicity of SCO was observed after treatment of strain CC104 with a concentration above its CMC, in which micelles were found. Nevertheless, this treatment was not cytotoxic to a CC104mutMmutY strain. Furthermore, in this DNA repair-deficient strain treatment with a SCO dose below its CMC, in which only monomers were found, demonstrated the possibility of an antioxidant effect, since a reduction in spontaneous mutagenesis frequency was observed. Finally, in an Ames test without metabolic activation mutagenicity induction was observed in strains TA100 and TA104 with treatment doses below the CMC. The cytotoxic, antioxidant and mutagenic effects of SCO can be influenced by the aggregational state. PMID- 15548755 TI - Determination of sediment mutagenicity and cytotoxicity in an area subjected to petrochemical contamination. AB - This study is an evaluation of the mutagenic and cytotoxic activity of sediments in Bom Jardim stream, one of the tributaries of the Cai River basin, Rio Grande do Sul, Brazil. This stream receives an indirect contribution of treated effluent from a petrochemical plant. The Salmonella/microsome assay, a microsuspension method, was used to evaluate moderately polar extracts of sediment samples at three points along the stream. The grain size analysis showed a lower mean content of fine particles in the principle face (front) of the complex, and this was also the sampling point with the lowest percentage of extracted organics. Low mutagenic activity was observed at the different sites studied, ranging from 3.3 to 8.3%; cytotoxic activity was more important in this area, ranging from 20 to 40%, adding up the results of assays in the presence and absence of external metabolism. In assays without S9mix there were more frequent mutagenic and cytotoxic responses, with frameshift mutations being the most frequent. The results also showed that there was a gradual, seasonal distribution of the responses as the stream mouth is reached, the most compromised points being in front of and downstream of the complex. Mutagenic and cytotoxic activity in sediment samples has proved important to determine environmental quality, despite the complexity of the chemical composition of the environmental matrix. Furthermore, use of the Salmonella assay to monitor mutagenesis and cytotoxicity helped identify the presence of pollutants. This assay is an important tool, aimed mainly at actions to preserve the genetic heritage of the fauna and flora affected by human activity and to improve environmental quality. PMID- 15548756 TI - Association between copper deficiency and DNA damage in cattle. AB - Cattle hypocuprosis is the second most widespread mineral deficiency affecting grazing cattle. The consequences of hypocuprosis include a failure of copper metalloenzymes, many of which form part of the antioxidant defence system. This work focuses on the association between copper (Cu) plasma concentration and DNA damage in Aberdeen Angus cattle. Two-hundred and ninety-nine heparinized blood samples from 2-year-old Aberdeen Angus cows were obtained from different farms located in the Salado River basin, Argentina. Plasma copper level analysis was carried out in whole samples, while cytogenetic analysis and single cell gel electrophoresis assay (comet assay) were carried out in 82 and 217 samples, respectively. Cytogenetic analysis showed a significant increase in the frequency of abnormal metaphases in moderate/severe hypocupremic groups (groups B and C) in relation to the normocupremic group (group A) (4.5 and 1.5 abnormal metaphases/100 cells, respectively, P < 0.01). The Spearman correlation test showed a negative association between cupremic values and the yield of chromosomal aberrations (r = -0.708, P < 0.0001). In the comet assay greater migration was observed in cells from the hypocupremic group, from a median of 54 in the severe hypocupremic group to 31 in the normocupremic group (P < 0.01). Accordingly, the Spearman correlation test showed a significant positive relationship between copper levels and cells without DNA migration and a significant negative relationship between copper levels and cells with a tiny tail (P < 0.0001 in both cases). The results obtained show that hypocupremia in cattle is associated with an increase in the frequency of chromosomal aberrations as well as in DNA migration as assessed by the comet assay. Whereas the comet assay could differentiate copper plasma level groups, chromosomal aberrations only detected differences between normal and hypocupremic animals. The increase of DNA damage found in hypocupremic animals could be explained by higher oxidative stress suffered by these animals. PMID- 15548757 TI - Measuring the genetic damage in cancer patients during radiotherapy with three genetic end-points. AB - In this paper the genetic damage for three genetic end-points was studied in cancer patients during radiotherapy using the micronucleus test, comet assay and hprt gene mutation test. The subjects were from: (i) a group of 24 patients suffering from various types of cancers; (ii) a group of 23 controls matched according to age, sex and smoking. Blood samples were collected from the controls and from the cancer patients prior to radiotherapy and after cumulative radiation doses of 10, 30 and 50 Gy. The results of the micronucleus test indicated that the mean micronuclei rate (MNR) and mean micronucleated cells rate (MCR) in the cancer patients prior to radiotherapy were 12.46 and 11.29 per thousand, respectively, which were significantly higher than those (6.65 and 5.96 per thousand) in controls (P < 0.01). However, the results of the comet assay and hprt gene mutation test showed no significant difference between untreated cancer patients and controls (P > 0.05). The mean MNR/MCR at 0, 10, 30 and 50 Gy in cancer patients increased with the cumulative dose, being 12.46, 35.83, 64.63 and 85.00 per thousand for MNR and 11.29, 31.25, 53.63 and 67.28 per thousand for MCR, respectively. Similar results were obtained in the comet assay and hprt gene mutation test. Meanwhile, it was found that there was inter-individual variability in response to radiotherapy among patients. PMID- 15548758 TI - DNA damaging effects of sulfur dioxide derivatives in cells from various organs of mice. AB - The DNA-damaging effects of sulfur dioxide (SO2) derivatives (a mixture of sodium sulfite and sodium bisulfite, 3:1 M/M) in the cells of various organs (brain, lung, heart, liver, stomach, spleen, thymus, bone marrow and kidney) of male mice were studied using the single cell gel electrophoresis technique (SCGE). Three groups of six mice each received an i.p. dose of SO2 derivatives (125, 250 or 500 mg/kg body wt) daily for 7 days. A control group of six mice received 200 microl of normal saline i.p. daily for 7 days. Our results show that SO2 derivatives caused significant increases in olive tail moment (OTM) in cells from all organs tested in a dose-dependent manner. These results show that SO2 derivatives can cause DNA damage to multiple organs of mice and that SO2 derivatives are systemic DNA-damaging agents, not only to the respiratory system. It is suggested that SO2 derivative exposure has a potential risk to DNA in multiple organs of mammals and might be related to carcinogenesis or other diseases related to DNA damage. Further work is required to understand the toxicological role of SO2 and its derivatives on multiple or even all organs in humans and animals. Recent research results have shown that SO2 and its derivatives can also induce an increase in the frequencies of chromosomal aberrations, sister chromatid exchanges and micronuclei in mammalian cells and cause oxidative damage in multiple organs of male and female mice. Taken together, these results suggest that SO2 and its derivatives are systemic toxic agents. However, further studies need to be performed before a definitive conclusion can be drawn. PMID- 15548759 TI - Folate levels determine effect of antioxidant supplementation on micronuclei in subjects with cardiovascular risk. AB - We have investigated the effect of modest supplementation with alpha-tocopherol (100 mg/day), beta-carotene (6 mg/day), vitamin C (100 mg/day) and selenium (50 microg/day) on oxidative stress and chromosomal damage, and the influence of methylenetetrahydrofolate reductase (MTHFR) genotype on these end-points. Subjects were two groups of middle-aged men differing in cardiovascular risk; 46 survivors of myocardial infarction before age 50 and 60 healthy controls. They were randomly divided into equal groups to receive antioxidants or placebo for 12 weeks. Twenty-eight patients and 58 controls completed the intervention. Micronucleus levels in peripheral lymphocytes and changes seen after intervention were studied in relation to the MTHFR C677T genotype, basal homocysteine and plasma folate levels. Ferric reducing ability of plasma and concentration of malondialdehyde were measured to assess the antioxidant effect of supplementation. There was no association of micronuclei with folate, homocysteine or malondialdehyde levels before supplementation. Micronucleus frequencies and plasma folate levels did not vary significantly with MTHFR genotype. Homocysteine levels in subjects with the TT variant genotype were significantly higher compared with CT or CC (P = 0.001), especially in subjects with low folate (P = 0.012). In the placebo control group an increase in micronuclei (P = 0.04) was detected at the end of the intervention period. This effect was not seen in the supplemented group. In antioxidant-supplemented myocardial infarction survivors we found an increase in the ferric reducing ability of plasma (P < 0.001) and a decrease in malondialdehyde (P = 0.001). Micronucleus frequency showed a decrease, strongest in subjects with normal folate levels (P = 0.015). In subjects with low folate levels, a high correlation was found between micronuclei after supplementation and homocysteine, both before (r = 0.979, P = 0.002) and after supplementation (r = 0.922, P = 0.009). Thus, folate deficiency may amplify the effect of other risk factors such as elevated homocysteine levels or variant MTHFR genotype, as well as influencing the ability of antioxidant supplementation to protect against genetic damage. PMID- 15548760 TI - Baseline levels of chromosome instability in the human lymphoblastoid cell TK6. AB - Induced genomic instability in the human B lymphoblastoid cell line TK6 manifests itself as increases in end-to-end chromosome fusions and non-reciprocal chromosome translocations. It is not associated with elevated frequencies of specific locus mutations or other cytogenetic alterations. Previous studies on a limited number of cells and end-points suggested that induced instability in TK6 mirrors spontaneous instability in terms of the types of alterations observed. In the present study we expanded on our previous analysis to include more cells and more end-points in order to derive a more precise measure of spontaneous instability in TK6 cells. The frequency of normal growth rate thymidine kinase mutants (TK(-/-)), measured in 44 independently isolated clones, was 2.73 +/- 0.78 x 10(-6)/cell, while that for slow growth mutants was 2.39 +/- 0.52 x 10( 6)/cell. These are similar to the frequencies observed for HPRT mutants in primary human cells. There was wide variation in chromatid break frequencies, but the average break frequency, at 0.04+/-0.01 breaks/cell, was only slightly higher than that reported for primary human cells. In contrast, the dicentric frequency of 0.006/cell was more than 10-fold higher for TK6 cells than that reported for normal primary human cells. Furthermore, the dicentrics in TK6 cells are unusual in that they are the result of end-to-end chromosome fusions. TK6 cells also show much higher levels of non-reciprocal chromosome translocations than are usually observed in primary human cells. The results suggest an inherent instability in TK6 cells that differs from what is observed in primary cells in that it affects the frequency of end-to-end chromosome fusions and non-reciprocal chromosome translocations, but not TK gene mutations or other cytogenetic alterations. PMID- 15548761 TI - DNA damage studies in untreated and treated leprosy patients. AB - The alkaline single cell gel electrophoresis assay was performed on peripheral blood lymphocytes of lepromatous and tubercloid leprosy patients (untreated and those undergoing treatment) in order to ascertain whether differential damage to DNA occurs. The study group included 28 male and 2 female patients and 15 healthy males who were matched for age and socio-economic status. The results revealed DNA damage in all patients, with a mean DNA migration length of 29.88 +/- 3.39 microm and 38% of their cells damaged when compared with the respective values obtained in healthy controls (1.28 +/- 0.40 microm, 5%). Multiple regression analysis for effects of confounding factors revealed antibiotic treatment in patients and alcohol consumption in controls as the only variables influencing DNA damage. In lepromatous and tubercloid patients, both untreated and those undergoing treatment, DNA damage increased significantly from that observed in control individuals, with greater increased damage in lepromatous patients. An increase in treatment time increased DNA damage linearly. Furthermore, an arbitrary classification of damaged cells (categories I-IV) was made based on observed tail lengths in leprosy patients (5.00-225.00 microm). The number of damaged cells in untreated patients was lower than in those undergoing treatment; the latter also had more cells with greater DNA migration lengths. There were no category III or IV cells in the control group. The results of the study therefore reveal that patients undergoing therapy had significantly greater DNA damage than untreated patients, indicating bacterial infection and drug therapy as the causal factors, since lepromatous-type disease is the more severe form with the patients having lower resistance to Mycobacterium leprae and requiring heavier and prolonged dosage of antibiotics. The study also corroborates that the assay offers an opportunity for correlating levels of therapy-induced DNA damage with administered dose and for modulating the dose-schedule so as to achieve lower levels of genotoxic damage. PMID- 15548762 TI - Styrene monomer does not induce unscheduled DNA synthesis in the mouse liver following inhalation exposure. AB - Styrene monomer is a commercially important chemical used extensively in the production of plastics. It has been shown to induce lung tumours in the mouse via the inhalation route. Styrene monomer has shown a low reactivity with DNA and also a lack of genotoxic response in the mouse lung in vivo. Together with the fact that the mouse lung tumours were late occurring and mostly benign, which suggest a promotional effect rather than initiation, these factors have led to a suggestion that the tumours may not be of genotoxic origin. The studies examining the genotoxicity of styrene monomer in vivo have to date been predominantly cytogenetic assessments, although low levels of DNA adducts have been reported in the mouse liver and lung. In order to extend this evaluation, a mouse liver unscheduled DNA synthesis study has been performed to assess the ability of styrene monomer to induce DNA damage/repair. The negative response observed in this assay is consistent with the theory that tumours observed in mouse oncogenicity studies are non-genotoxic in origin. PMID- 15548763 TI - The use of IL-2 cultures to measure chromosomal radiosensitivity in breast cancer patients. AB - Enhanced chromosomal radiosensitivity in breast cancer patients has been demonstrated in several studies. To investigate the chromosomal radiosensitivity of lymphocytes in breast cancer patients the G2 and micronucleus (MN) assays are often used. In these assays blood samples are exposed to ionizing radiation and the number of radiation-induced micronuclei or chromatid breaks are scored. In most studies investigating the in vitro chromosomal radiosensitivity of breast cancer patients the G2 and MN assays were performed on freshly drawn blood. The disadvantage of working with fresh blood samples is that in most cases only one blood sample can be obtained and that the assay cannot be easily repeated without further blood sampling. To allow repeated testing we propose the use of long-term cultures of T lymphocytes (IL-2 cultures). In this study we therefore investigated whether the radiation-induced MN response in IL-2 cultures was the same as in concordant whole blood cultures. For this study the MN assay (2 Gy) was performed on IL-2 cultures of 11 sensitive breast cancer patients and 20 healthy women. The results demonstrate that the enhanced chromosomal radiosensitivity observed in whole blood cultures of breast cancer patients is not present in IL-2 cultures derived from the same blood samples. Therefore, care has to be taken when IL-2 cultures are used to assess chromosomal radiosensitivity in breast cancer patients. PMID- 15548764 TI - Discovery of a small molecule activator of the human ether-a-go-go-related gene (HERG) cardiac K+ channel. AB - Many drugs inhibit the human ether-a-go-go-related gene (HERG) cardiac K+ channel. This leads to action potential prolongation on the cellular level, a prolongation of the QT interval on the electrocardiogram, and sometimes cardiac arrhythmia. To date, no activators of this channel have been reported. Here, we describe the in vitro electrophysiological effects of (3R,4R)-4-[3-(6 methoxyquinolin-4-yl)-3-oxo-propyl]-1-[3-(2,3,5-trifluoro-phenyl)-prop-2-ynyl] piperidine-3-carboxylic acid (RPR260243), a novel activator of HERG. Using patch clamp electrophysiology, we found that RPR260243 dramatically slowed current deactivation when applied to cells stably expressing HERG. The effects of RPR260243 on HERG channel deactivation were temperature- and voltage-dependent and occurred over the concentration range of 1 to 30 microM. RPR260243-modified HERG currents were inhibited by dofetilide (IC50 = 58 nM). RPR260243 had little effect on HERG current amplitude and no significant effects on steady-state activation parameters or on channel inactivation processes. RPR260243 displayed no activator-like effects on other voltage-dependent ion channels, including the closely related erg3 K+ channel. RPR260243 enhanced the delayed rectifier current in guinea pig myocytes but, when administered alone, had little effect on action potential parameters in these cells. However, RPR260243 completely reversed the action potential-prolonging effects of dofetilide in this preparation. Using the Langendorff heart method, we found that 5 microM RPR260243 increased T-wave amplitude, prolonged the PR interval, and shortened the QT interval. We believe RPR260243 represents the first known HERG channel activator and that the drug works primarily by inhibiting channel closure, leading to a persistent HERG channel current upon repolarization. Compounds like RPR260243 will be useful for studying the physiological role of HERG and may one day find use in treating cardiac disease. PMID- 15548765 TI - N3-methyladenine induces early poly(ADP-ribosylation), reduction of nuclear factor-kappa B DNA binding ability, and nuclear up-regulation of telomerase activity. AB - Methylation of N3-adenine represents a novel pharmacological strategy for the treatment of resistant tumors. However, little is known about the biochemical pathways involved in cell death induced by N3-methyladenine. In the present study, we show that MeOSO(2) (CH(2))(2)-lexitropsin (Me-Lex), a compound generating almost exclusively N3-methyladenine (>99%), provoked a burst of poly(ADP-ribosylation) and loss of mitochondrial membrane potential in leukemia cells. These events were followed by a marked decrease in nuclear poly(ADP ribose) polymerase-1 (PARP-1) expression and nuclear factor-kappaB (NF-kappaB) activity. Moreover, DNA damage generated by N3-methyladenine induced a marked decrease in telomerase in the cytosol that was accompanied by a transient up regulation of activity in the nucleus, as a consequence of nuclear translocation of telomerase in response to genotoxic damage. PARP-1 inhibition blocked ADP ribose polymer formation, preserved mitochondrial membrane integrity, and counteracted the reduction of NF-kappaB activity, thus preventing the appearance of necrosis. On the other hand, because PARP-1 is a component of the base excision repair (BER), the combination of Me-Lex + PARP-1 inhibitor triggered apoptosis as a result of disruption of BER process. In conclusion, the present study provides new insight into the cellular response to N3-adenine-selective methylating agents that can be exploited for the treatment of tumors unresponsive to classical wide-spectrum methylating agents. Moreover, the results underline the central and paradoxical role of PARP-1 in cell death induced by N3 methyladenine: effector of necrosis and coordinator of methylpurine repair. PMID- 15548766 TI - The mGluR5 antagonist 6-methyl-2-(phenylethynyl)pyridine decreases ethanol consumption via a protein kinase C epsilon-dependent mechanism. AB - Glutamatergic neurotransmission plays a critical role in addictive behaviors, and recent evidence indicates that genetic or pharmacological inactivation of the type 5 metabotropic glutamate receptor (mGluR5) reduces the self-administration of cocaine, nicotine, and alcohol. Because mGluR5 is coupled to activation of protein kinase C (PKC), and targeted deletion of the epsilon isoform (PKCepsilon) in mice reduces ethanol self-administration, we investigated whether there is a functional link between mGluR5 and PKCepsilon. Here, we show that acute administration of the mGluR5 agonist (R,S)-2-chloro-5-hydroxyphenylglycine to mice increases phosphorylation of PKCepsilon in its activation loop (T566) as well as in its C-terminal region (S729). Increases in phospho-PKCepsilon are dependent not only on mGluR5 stimulation but also on phosphatidylinositol-3 kinase (PI3K). In addition, the selective mGluR5 antagonist 6-methyl-2 (phenylethynyl)pyridine (MPEP) reduced basal levels of phosphorylation of PKCepsilon at S729. We also show that MPEP dose dependently reduced ethanol consumption in wild-type but not in PKCepsilon-null mice, suggesting that PKCepsilon is an important signaling target for modulation of ethanol consumption by mGluR5 antagonists. Radioligand binding experiments using [(3)H]MPEP revealed that these genotypic differences in response to MPEP were not a result of altered mGluR5 levels or binding in PKCepsilon-null mice. Our data indicate that mGluR5 is coupled to PKCepsilon via a PI3K-dependent pathway and that PKCepsilon is required for the ability of the mGluR5 antagonist MPEP to reduce ethanol consumption. PMID- 15548767 TI - Solving the energy crisis. Signaling molecule spurs cells to generate mitochondria. PMID- 15548768 TI - Ironing out cell death.Metal-grabbing enzyme saves suicidal cells. PMID- 15548769 TI - Culture clash.A growing body of research suggests that yeast have programmed death pathways, yet many researchers are skeptical. Recent studies provide some of the first experimental evidence for why a single-celled organism would commit suicide. PMID- 15548770 TI - Images in clinical medicine. Left nasal discharge and meningitis. PMID- 15548771 TI - Merck's recall of rofecoxib--a strategic perspective. PMID- 15548772 TI - House calls. PMID- 15548773 TI - Prognosis in follicular lymphoma--it's in the microenvironment. PMID- 15548774 TI - Ultrasound energy and the dissolution of thrombus. PMID- 15548775 TI - Prior-authorization programs for controlling drug spending. PMID- 15548776 TI - Prediction of survival in follicular lymphoma based on molecular features of tumor-infiltrating immune cells. AB - BACKGROUND: Patients with follicular lymphoma may survive for periods of less than 1 year to more than 20 years after diagnosis. We used gene-expression profiles of tumor-biopsy specimens obtained at diagnosis to develop a molecular predictor of the length of survival. METHODS: Gene-expression profiling was performed on 191 biopsy specimens obtained from patients with untreated follicular lymphoma. Supervised methods were used to discover expression patterns associated with the length of survival in a training set of 95 specimens. A molecular predictor of survival was constructed from these genes and validated in an independent test set of 96 specimens. RESULTS: Individual genes that predicted the length of survival were grouped into gene-expression signatures on the basis of their expression in the training set, and two such signatures were used to construct a survival predictor. The two signatures allowed patients with specimens in the test set to be divided into four quartiles with widely disparate median lengths of survival (13.6, 11.1, 10.8, and 3.9 years), independently of clinical prognostic variables. Flow cytometry showed that these signatures reflected gene expression by nonmalignant tumor-infiltrating immune cells. CONCLUSIONS: The length of survival among patients with follicular lymphoma correlates with the molecular features of nonmalignant immune cells present in the tumor at diagnosis. PMID- 15548777 TI - Ultrasound-enhanced systemic thrombolysis for acute ischemic stroke. AB - BACKGROUND: Transcranial Doppler ultrasonography that is aimed at residual obstructive intracranial blood flow may help expose thrombi to tissue plasminogen activator (t-PA). Our objective was to determine whether ultrasonography can safely enhance the thrombolytic activity of t-PA. METHODS: We treated all patients who had acute ischemic stroke due to occlusion of the middle cerebral artery with intravenous t-PA within three hours after the onset of symptoms. The patients were randomly assigned to receive continuous 2-MHz transcranial Doppler ultrasonography (the target group) or placebo (the control group). The primary combined end point was complete recanalization as assessed by transcranial Doppler ultrasonography or dramatic clinical recovery. Secondary end points included recovery at 24 hours, a favorable outcome at three months, and death at three months. RESULTS: A total of 126 patients were randomly assigned to receive continuous ultrasonography (63 patients) or placebo (63 patients). Symptomatic intracerebral hemorrhage occurred in three patients in the target group and in three in the control group. Complete recanalization or dramatic clinical recovery within two hours after the administration of a t-PA bolus occurred in 31 patients in the target group (49 percent), as compared with 19 patients in the control group (30 percent; P=0.03). Twenty-four hours after treatment of the patients eligible for follow-up, 24 in the target group (44 percent) and 21 in the control group (40 percent) had dramatic clinical recovery (P=0.7). At three months, 22 of 53 patients in the target group who were eligible for follow-up analysis (42 percent) and 14 of 49 in the control group (29 percent) had favorable outcomes (as indicated by a score of 0 to 1 on the modified Rankin scale) (P=0.20). CONCLUSIONS: In patients with acute ischemic stroke, continuous transcranial Doppler augments t-PA-induced arterial recanalization, with a nonsignificant trend toward an increased rate of recovery from stroke, as compared with placebo. PMID- 15548778 TI - Premature birth and later insulin resistance. AB - BACKGROUND: Term infants who are small for gestational age appear prone to the development of insulin resistance during childhood. We hypothesized that insulin resistance, a marker of type 2 diabetes mellitus, would be prevalent among children who had been born prematurely, irrespective of whether they were appropriate for gestational age or small for gestational age. METHODS: Seventy two healthy prepubertal children 4 to 10 years of age were studied: 50 who had been born prematurely (32 weeks' gestation or less), including 38 with a birth weight that was appropriate for gestational age (above the 10th percentile) and 12 with a birth weight that was low (i.e., who were small) for gestational age, and 22 control subjects (at least 37 weeks' gestation, with a birth weight above the 10th percentile). Insulin sensitivity was measured with the use of paired insulin and glucose data obtained by frequent measurements during intravenous glucose-tolerance tests. RESULTS: Children who had been born prematurely, whether their weight was appropriate or low for gestational age, had an isolated reduction in insulin sensitivity as compared with controls (appropriate-for gestational-age group, 14.2x10(-4) per minute per milliunit per liter [95 percent confidence interval, 11.5 to 16.2]; small-for-gestational-age group, 12.9x10(-4) per minute per milliunit per liter [95 percent confidence interval, 9.7 to 17.4]; and control group, 21.6x10(-4) per minute per milliunit per liter [95 percent confidence interval, 17.1 to 27.4]; P=0.002). There were no significant differences in insulin sensitivity between the two premature groups (P=0.80). As compared with controls, both groups of premature children had a compensatory increase in acute insulin release (appropriate-for-gestational-age group, 2002 pmol per liter [95 percent confidence interval, 1434 to 2432] [corrected]; small for-gestational-age group, 2253 pmol per liter [95 percent confidence interval, 1622 to 3128]; and control group, 1148 pmol per liter [95 percent confidence interval, 875 to 1500]; P<0.001). CONCLUSIONS: Like children who were born at term but who were small for gestational age, children who were born prematurely have an isolated reduction in insulin sensitivity, which may be a risk factor for type 2 diabetes mellitus. PMID- 15548779 TI - Medicaid prior-authorization programs and the use of cyclooxygenase-2 inhibitors. AB - BACKGROUND: Over the past five years, selective cyclooxygenase-2 inhibitors (coxibs) have accounted for a growing proportion of prescriptions for nonsteroidal antiinflammatory drugs (NSAIDs). To control these expenses, many state Medicaid programs have implemented prior-authorization requirements before coxibs can be prescribed. We evaluated the effect of such programs on the use of coxibs by Medicaid beneficiaries. METHODS: We surveyed state Medicaid agencies to determine whether prescription of coxibs required prior authorization and, if so, the criteria for authorization. For each program, we compared these criteria with evidence-based recommendations for prescribing of coxibs. Using data for all filled prescriptions in 50 state Medicaid programs from 1999 through the end of 2003, we calculated the proportion of defined daily doses of NSAIDs accounted for by coxibs. Time-series analyses were used to measure the changes in prescription patterns after the implementation of each prior-authorization program. RESULTS: By 2001, coxibs accounted for half of all NSAID doses covered by Medicaid. This proportion varied widely according to the state in 2003, from a low of 11 percent to a high of 70 percent of all NSAID doses. Twenty-two states implemented prior authorization programs for coxibs during the study period. Overall, the implementation of such programs reduced the proportion of NSAID doses made up by coxibs by 15.0 percent (95 percent confidence interval, 10.9 to 19.2 percent), corresponding to a decrease of 10.28 dollars (95 percent confidence interval, 7.56 dollars to 13.00 dollars) in spending per NSAID prescription. The effect of such programs was not influenced by the degree to which a prior-authorization program incorporated evidence-based prescribing recommendations. CONCLUSIONS: The use of coxibs and spending on NSAIDs varies widely by state and declined substantially after the implementation of prior-authorization programs. Determining whether these reductions are clinically appropriate will have important implications for the development of rational drug-reimbursement policies. PMID- 15548780 TI - Clinical practice. Acute pericarditis. PMID- 15548781 TI - Advances in H1-antihistamines. PMID- 15548782 TI - Images in clinical medicine. Scleromyxedema. PMID- 15548783 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 35-2004. A 68-year-old man with end-stage renal disease and thickening of the skin. PMID- 15548784 TI - Prematurity--a window of opportunity? PMID- 15548785 TI - HIV-1 and HCV infections among antibody-negative blood donors. PMID- 15548786 TI - MRI in breast cancer. PMID- 15548787 TI - C/EBPalpha in asthma. PMID- 15548788 TI - Hypocalciuric hypercalcemia and autoantibodies against the calcium-sensing receptor. PMID- 15548789 TI - More on blue cohosh and perinatal stroke. PMID- 15548790 TI - Generalized anxiety disorder. PMID- 15548791 TI - Soluble fms-like tyrosine kinase 1. PMID- 15548792 TI - Uveitis and neurological diseases. PMID- 15548793 TI - Threshold ROP. PMID- 15548794 TI - Found in translation. PMID- 15548795 TI - A randomised trial of povidone-iodine to reduce visual impairment from corneal ulcers in rural Nepal. AB - AIM: To assess whether povidone-iodine provided any benefit over and above a standard regimen of antibiotic therapy for the treatment of corneal ulcers. METHODS: All patients diagnosed with corneal ulcers presenting for care at a primary eye care clinic in rural Nepal were randomised to a standard protocol of antibiotic therapy versus standard therapy plus 2.5% povidone-iodine every 2 hours for 2 weeks. The main outcomes were corrected visual acuity and presence, size, and position of corneal scarring in the affected eye at 2-4 months following treatment initiation. RESULTS: 358 patients were randomised and 81% were examined at follow up. The two groups were comparable before treatment. At follow up, 3.9% in the standard therapy and 6.9% in the povidone-iodine group had corrected visual acuity worse than 20/400 (relative risk (RR) 1.77, 95% confidence interval (CI) 0.62 to 5.03). 9.4% in the standard therapy and 13.1% in the povidone-iodine group had corrected visual acuity worse than 20/60 (RR 1.39, 95% CI 0.71 to 2.77), and 17.0% and 18.8% had scars in the visual axis in each of these groups, respectively (RR 1.11, 95% CI 0.67 to 1.82). CONCLUSIONS: A small proportion of patients with corneal ulceration treated in this setting had poor visual outcomes. The addition of povidone-iodine to standard antibiotic therapy did not improve visual outcomes, although this design was unable to assess whether povidone-iodine on its own would have resulted in comparable visual outcomes to that of standard therapy. PMID- 15548796 TI - The prevalence of glaucoma in Bangladesh: a population based survey in Dhaka division. AB - BACKGROUND/AIM: Few studies have reported on the epidemiology and clinical characteristics of glaucoma in south Asia. This project aimed to determine the prevalence of glaucoma of Dhaka, Bangladesh. METHOD: A multistage, stratified, clustered sample was drawn from Dhaka Division, Bangladesh, using systematic sampling to identify individual subjects aged 35 years and older. Examination of all subjects included Snellen visual acuity, slit lamp examination (including gonioscopy and applanation tonometry) and a stereoscopic assessment of the vertical cup:disc ratio (CDR). In selected subjects, a threshold visual field examination was performed. Glaucoma was diagnosed on the basis of statistical abnormality of the vertical CDR combined with an abnormal visual field test, or in subjects with advanced glaucoma who could not complete field testing, a grossly abnormal CDR. If it was not possible to examine the optic discs and the subject was blind, glaucoma was diagnosed on the basis of a raised intraocular pressure. RESULTS: Of 3562 eligible subjects, 2347 were examined (66%). Among people aged 40 years and older, the prevalence of definite glaucoma was 2.1% (95% confidence interval: 1.5 to 2.9; 39 people). The prevalence of definite and probable glaucoma was 3.1% (95% CI: 2.4 to 4.0; 58 people) in subjects of the same age. Primary open angle glaucoma was the most common form of glaucoma, accounting for 75% of the total. Among cases of blindness not attributable to refractive error, 5% were caused by glaucoma. Glaucoma prevalence was higher in men than women, but did not show the typical increase with age. CONCLUSION: Glaucoma prevalence is relatively high in Bangladesh, although it accounts for only a small proportion of blindness in the community. It is estimated that there are approximately 586 000 people 40 years and older with definite or probable glaucoma in Bangladesh. PMID- 15548797 TI - Neurological concomitants of uveitis. AB - AIM: To describe the prevalence and types of neurological disease that occur in association with uveitis. METHODS: Retrospective review of medical records of patients attending a tertiary referral uveitis service over a 15 year period. RESULTS: Of 1450 patients with uveitis, 115 (7.9%) had neurological disease that was considered to be causally related to the eye inflammation. The most frequent neurological associations were Vogt-Koyanagi-Harada disease, primary central nervous system lymphoma, multiple sclerosis, and herpes virus infections. CONCLUSIONS: Neurological disease is common among patients attending a uveitis service. The distinctive characteristics of the uveal inflammation may be useful in diagnosing the neurological disease. PMID- 15548798 TI - Variations in the morphology of retinopathy of prematurity in extremely low birthweight infants. AB - AIMS: To investigate the clinical observations that arteriovenous shunts typical of threshold retinopathy of prematurity (ROP) are morphologically different in extremely low birthweight infants weighing less than 1000 g. METHODS: An observational case series of six extremely low birthweight infants displaying specific features of threshold retinopathy of prematurity enrolled between 1998 and 2001 at one centre. The variant morphology was documented with colour photography and fundus fluorescein angiography before laser therapy. RESULTS: Stage 3 threshold ROP in extremely premature infants may be characterised by a different morphology not demonstrating classic shunt formation. A poorly developed capillary bed is present in already vascularised retina in these cases. CONCLUSIONS: This case series of extremely low birthweight infants display variations in the typical morphological appearance of threshold ROP. In these cases, established plus disease may be present in the absence of arteriovenous shunting. Delaying treatment until a classic stage 3 ridge with extraretinal neovascularisation develops may be detrimental to controlling the disease process. The authors propose that the criteria for threshold disease requiring treatment do not accurately apply in this extremely low birthweight group as defined by the CRYO-ROP study and that treatment should be instituted before the typical threshold features arise. Plus disease remains the most reliable sign indicating the need for treatment. PMID- 15548799 TI - Inflammatory cells in brush cytology samples correlate with the severity of corneal lesions in atopic keratoconjunctivitis. AB - BACKGROUND: Inflammatory cells infiltrating to the tarsal conjunctiva are thought to be involved in the pathogenesis of corneal lesions in severe allergic conjunctival diseases. The relation between such cells and the severity of corneal lesions was studied. METHODS: Six patients with atopic keratoconjunctivitis (AKC) were enrolled in this study. Tarsal brush cytology findings and the severity of corneal damage at that point were recorded and analysed for correlation. RESULTS: Four out of six patients exhibited correlation between eosinophils and corneal damage. Three out of six patients exhibited correlation between neutrophils and corneal damage. Two out of six patients exhibited correlation between both eosinophils and neutrophils and corneal damage. Analysis of all data from all patients taken together revealed that both eosinophils and neutrophils in brush cytology samples significantly correlated with corneal damage. CONCLUSIONS: Inflammatory cells in brush cytology samples correlated with corneal damage. Evaluation of the relative percentages of inflammatory cells in brush cytology samples is a useful method of assessing disease activity in allergic conjunctival disease. PMID- 15548800 TI - A correlation of pregnancy term, disease activity, serum female hormones, and cytokines in uveitis. AB - BACKGROUND/AIMS: Pregnancy and the postpartum period are associated with the activity of autoimmune diseases including uveitis. Although the exact mechanism is unknown, hormones are reported to alter inflammatory cytokines and influence disease activity. The authors studied ocular inflammation, female hormones, and serum cytokine levels during and after pregnancy. METHODS: A prospective, observational case study was conducted. Four pregnant women in their first trimester with chronic non-infectious uveitis were followed monthly until 6 months after delivery. Serum female hormones (oestrogen, progesterone, prolactin) and various cytokines (IL-2, IL-4, IL-5, IL-6, IL-10, IFN-gamma, and TGF-beta) were measured by ELISA. RESULTS: The four patients had five full term pregnancies. Uveitis activity decreased after the first trimester but flared in the early postpartum period. Serum female hormones, highly elevated during pregnancy, drastically dropped post partum. Cytokine levels except TGF-beta were mostly undetectable. CONCLUSION: Female hormones and TGF-beta may contribute to the activity of uveitis during pregnancy and the postpartum period. PMID- 15548801 TI - Impact factors on intraocular pressure measurements in healthy subjects. AB - AIM: To evaluate whether intraocular pressure (IOP) calculation by applanation tonometry is determined more essentially by the subject's neck position or by neck constriction. METHODS: 23 right eyes of 23 healthy subjects (12 male, 11 female) were included. IOP was measured by applanation tonometry with the TonoPen on sitting participants under four different conditions: with open collar upright (A) or with the head in the headrest of a slit lamp (B), with a tight necktie upright (C) or in slit lamp position (D). All measurements with neck constriction were performed 3 minutes after placing the necktie. RESULTS: Mean IOP was 16.9 (SD 2.3) mm Hg (range 11-21 mm Hg) (A), 18.1 (SD 2.2) mm Hg (range 14-22 mm Hg) (B), 17.9 (SD 2.9) mm Hg (range 12-25 mm Hg) (C) and 18.7 (SD 2.7) mm Hg (range 13-24 mm Hg) (D). Mean IOP increased by 1.3 (SD 2.6) mm Hg (p = 0.028, paired t test, range +0.2 to +2.4 mm Hg) if subjects changed position from A to B. There was no statistically significant difference between measurements with or without neck constriction. CONCLUSION: Applanation tonometry may be inaccurate if performed in slit lamp position. In contrast, tight neckties do not significantly affect IOP evaluation in healthy subjects. PMID- 15548802 TI - Cataract surgery and subtype in a defined, older population: the SEECAT Project. AB - AIM: To describe the distribution of cataract subtypes present before surgery among a defined population of older, bilaterally pseudophakic individuals. METHODS: This was a cohort study of bilaterally pseudophakic individuals participating in the Salisbury Eye Evaluation (SEE), and their locally resident siblings. Subjects underwent slit lamp and retroillumination photography and grading using the Wilmer Cataract Grading System. For all individuals determined to be bilaterally pseudophakic, an attempt was made to determine for each eye the type(s) of cataract present before surgery, based on previous SEE photographs (for SEE participants) and/or medical records obtained from the operating ophthalmologist (for both SEE participants and their siblings). RESULTS: The mean age of 223 participants providing data in this study was 78.7 (SD 5.2) years, 19.3% of subjects were black and 60.1% female. The most common surgically removed cataract subtype in this population was pure nuclear (43.5%), followed by nuclear combined with posterior subcapsular cataract (PSC) (20.6%), and nuclear combined with cortical (13.9%); less common types were pure cortical (4.9%), pure PSC (4.5%), and PSC combined with cortical (2.7%). Factors such as sex and source of lens data (study photograph versus clinical record) did not significantly affect the distribution of lens opacity types, while PSC was significantly (p = 0.01) more common among younger people and nuclear cataract was significantly (p = 0.001) more common among white compared to black people. CONCLUSION: Epidemiological studies have suggested that the different subtypes of cataract are associated with different risk factors. As studies begin to identify new prevention strategies for cataract, it would appear likely that different strategies will be efficacious against different types of cataract. In this setting, it will be helpful to know which cataract types are most frequently associated with surgery. Among this older, majority white population, nuclear cataract showed a clear predominance among individuals having undergone surgery in both eyes. This may be contrasted with both clinic and population based studies of younger people, which have generally found PSC cataract to predominate. PMID- 15548803 TI - Is screening for interferon retinopathy in hepatitis C justified? AB - BACKGROUND/AIM: In the treatment of hepatitis C, the National Institute for Clinical Excellence advocates use of a combination of interferon alfa and ribavirin for selected patients. Retinopathy is a well recognised side effect of interferon therapy and is characterised by retinal haemorrhages, cotton wool spots, and macular oedema. The aim of this study was to document the incidence and natural history of the retinopathy in patients treated with a long acting (pegylated) interferon and ribavirin for hepatitis C and to assess the need to screen for retinal complications. METHODS: All patients started on treatment from September 2002 to August 2003 were invited to participate in the study. The past medical and ocular history, visual symptoms, and the results of a full ophthalmological assessment performed 3 months after starting treatment were noted. Any patient with retinal changes was followed up at 3 month intervals until the changes resolved. RESULTS: Of the 25 patients examined, four had evidence of retinopathy including deep retinal haemorrhage and cotton wool spots. Two of the patients were diabetic and one hypertensive. None had any visual symptoms and in all four the retinopathy resolved while the patients completed their course of treatment. CONCLUSIONS: The incidence of retinopathy with pegylated interferon is low. The retinal complications resolve while treatment is continued and are asymptomatic. This study does not support routine screening for retinopathy in patients treated with pegylated interferon and ribavirin for hepatitis C. PMID- 15548804 TI - Multidrug resistant proteins: P-glycoprotein and lung resistance protein expression in retinoblastoma. AB - BACKGROUND/AIM: Retinoblastoma is the commonest primary intraocular tumour in children. Chemotherapy now plays a big part in the treatment of these tumours. There is not much information about the role of the multidrug resistance proteins (MDR)-P-glycoprotein (P-gp) and vault protein lung resistance protein (LRP)-in retinoblastoma. The authors investigated the expression of P-gp and LRP in retinoblastoma and correlated them clinicopathologically. METHODS: Among 60 retinoblastomas, 40 tumours were not subjected to preoperative or postoperative chemotherapy and 20 tumours were subjected to postoperative chemotherapy. In this cohort 27 tumours had no invasion and 33 tumours had invasion of choroid, optic nerve, and orbit. P-gp and LRP expression were studied by immunohistochemistry. Immunoanalysis was done semiquantitatively. RESULTS: Among the 60 tumours P-gp was expressed in 23 (38%) tumours and LRP was expressed in 35 (58%). P-gp was expressed in 11/27 (40%) tumours with no invasion and in 12/33 (36%) tumours with invasion. LRP was expressed in 15/27 (55%) tumours with no invasion and in 20/33 (60%) tumours with invasion. Both P-gp and LRP were negative in three tumours with invasion, which had later developed bone marrow metastasis. There was no correlation between P-gp and LRP expression with invasion, differentiation and laterality of the tumours and response to treatment. CONCLUSION: Retinoblastoma expresses P-gp and LRP intrinsically before chemotherapy and none of these proteins predicted the response to chemotherapy. Thus, further studies are needed to understand the significance of the expression of the P-gp and LRP proteins in retinoblastoma. PMID- 15548805 TI - Identification of monosomy 3 in choroidal melanoma by chromosome in situ hybridisation. AB - BACKGROUND/AIMS: In uveal melanoma monosomy 3 is emerging as a significant indicator of a poor prognosis. To date most cytogenetic studies of uveal melanoma have utilised fresh tissue or DNA extracted from tissue sections. In this study chromosome in situ hybridisation (CISH) was used to study monosomy 3 in tissue sections. The copy number of chromosome 3 was determined and related to patient survival. METHODS: Archival glutaraldehyde or formalin fixed, paraffin embedded material was obtained from 30 metastasising and 26 non-metastasising choroidal melanomas. Hybridisations were performed using centromere specific probes to chromosomes 3 and 18. Chromosome 18 was included as a control as previous abnormalities in uveal melanoma have not been described. Chromosomal imbalance was defined on the basis of changes in both chromosome index and signal distribution. RESULTS: CISH was successfully performed on both glutaraldehyde and formalin fixed tissue. Four cases were unsuccessful because of extensive tumour necrosis. All cases were balanced for chromosome 18. Monosomy 3 was detected in 15 of the 26 cases of metastasising melanoma; the 26 non-metastasising tumours were all balanced for chromosome 3. Monosomy 3 was significantly associated with metastases related death. CONCLUSION: CISH can successfully identify monosomy 3 in archival glutaraldehyde or formalin fixed, paraffin embedded tissue sections. Similar to previous studies monosomy 3 is a significant predictor of metastases related death. PMID- 15548806 TI - Genotype-phenotype correlation in a family with Arg135Leu rhodopsin retinitis pigmentosa. AB - AIM: To describe the clinical characteristics and disease course of a large family with retinitis pigmentosa (RP) from an Arg135Leu change in rhodopsin. METHODS: 29 patients in this family were evaluated. Goldmann visual fields were performed on 14 affected individuals, Ganzfeld electroretinography (ERG) on eight individuals (11-56 years), and blood samples collected on 10 individuals (11-58 years). Patient visual field data were compared with previously reported patients with different rhodopsin mutations using linear regression. RESULTS: An Arg135Leu mutation was identified in rhodopsin. Distinct stages of clinical evolution were identified for this family ranging from normal, white dots, classic bone spicules and, finally, ending with extensive retinal pigment epithelium (RPE) atrophy. 9/16 patients over the age of 20 years also demonstrated marked macular atrophy. All patients who underwent full field ERG testing demonstrated non-recordable ERGs. The overall regression model comparing solid angles of visual fields from patients with rhodopsin mutations (Pro23His, Pro347Ala, Arg135Leu) shows significant effects for age (p = 0.0005), mutation (p = 0.0014), and interaction between age and mutation (p = 0.018) with an R(2) of 0.407. CONCLUSIONS: An Arg135Leu change in rhodopsin results in a severe form of RP that evolves through various fundus appearances that include white dots early in life and classic appearing RP later. This transmembrane change in rhodopsin proves to be more severe than in a family with an intradiscal change and a family with a cytoplasmic change. PMID- 15548807 TI - Elevated plasma levels of interleukin 8 in patients with acute anterior ischaemic optic neuropathy. AB - BACKGROUND/AIM: Alterations of the immune system may have a role in thrombogenesis. Artery sites occluded with thrombi apparently release pro inflammatory cytokines. Non-arteritic anterior ischaemic optic neuropathy (NAION) results from occlusion of the blood supply to the optic nerve. The aim of this study was to analyse levels of pro-inflammatory cytokines in patients with acute event of NAION. METHODS: Study participants included 10 patients (12 eyes) with NAION and 20 age matched controls with the same risk factors for atherosclerosis disease. Peripheral blood samples were obtained immediately at the acute onset of NAION. Plasma interleukin 8 (IL-8), IL-6, and tumour necrosis factor alpha (TNF alpha) levels were measured immediately following diagnosis and during the follow up intervals. RESULTS: The plasma levels of IL-8 were significantly higher in NAION patients at the time of diagnosis in comparison to the control group (p = 0.002), and decreased during the follow up period (6-12 months) (p = 0.05). There were no differences in plasma levels of IL-6 and TNF-alpha between NAION patients and controls, either in the acute phase or during the follow up period. CONCLUSION: Plasma levels of IL-8 are elevated during the acute phase of NAION, but not IL-6 and TNF-alpha. These elevated levels are in accordance with other acute vascular thrombosis. The clinical significance of these findings should be further evaluated. PMID- 15548808 TI - Visual acuity and the ability of the visually impaired to read medication instructions. AB - BACKGROUND/AIMS: Although medical information is often communicated in writing, little attention is given as to whether patients can read it--for example, the miniscule writing on the side of the medication bottles (equivalent to 6/6 reduced Snellen). The authors wished to determine a distance visual acuity threshold, using routine assessments of visual acuity (VA), beyond which patients are unable to read their instructions. METHODS: 180 patients, subgrouped according to the best line of Snellen acuity (from 6/9 to 6/60) achieved in either eye were recruited. Subjects were asked to read the printed manufacturer's instructions on the side of the drops box without magnification. RESULTS: Those subjects with best corrected VA of lower than or equal to 6/24 showed a significantly diminished ability to read the instructions on their eye drops bottles (p<0.001 for each comparison). When this group selected a font size of their choosing, the mode for preferred Arial font sizes were 16 for the 6/24 group, 18 for 6/36, and 22 for 6/60. CONCLUSION: This study documents the inability of patients with visual impairment to read the instructions on their bottle of eye drops. There is evidence of a distance VA threshold effect such that, although the majority of patients with 6/18 VA are able to read their instructions, patients whose VA is worse cannot. This latter group would benefit from a larger font size, a size of Arial 22 being sufficient. We recommend that all printed information for patients with visual acuities between 6/24 and 6/60 be printed in font Arial 22 or equivalent. PMID- 15548809 TI - Plasma vascular endothelial growth factor, angiopoietin-2, and soluble angiopoietin receptor tie-2 in diabetic retinopathy: effects of laser photocoagulation and angiotensin receptor blockade. AB - BACKGROUND: Proliferative diabetic retinopathy (PDR) may be a response to abnormal angiogenic growth factors such as vascular endothelial growth factor (VEGF), angiopoietin-2 (Ang-2), and the soluble angiopoietin receptor tie-2. The authors hypothesised the following: (a) there are differences in plasma levels of these growth factors in different grades of diabetic retinopathy; and (b) that the effects of intervention with panretinal laser photocoagulation (PRP) for PDR, and angiotensin receptor blockade (using eprosartan) for patients with other grades of diabetic retinopathy will be to reduce levels of the growth factors. METHODS: Cross sectional and interventional study (using PRP and eprosartan) in diabetic patients. VEGF, Ang-2, and tie-2 were measured by ELISA. RESULTS: VEGF (p<0.001) and Ang-2 levels (p<0.001) were significantly higher in 93 diabetic patients compared to 20 healthy controls, with the highest levels in grade 2 and grade 3 diabetic retinopathy (p<0.05). Tie-2 was lower in diabetics compared to controls (p = 0.008), with no significant differences between the diabetic subgroups. Overall, VEGF significantly correlated with Ang-2 (p<0.001) and tie-2 (p = 0.004) but the correlation between Ang-2 and tie-2 levels was not significant (p = 0.065). Among diabetic patients only, VEGF levels were significantly correlated with Ang-2 (p<0.001) and tie-2 (p<0.001); the correlation between Ang-2 and tie-2 levels was also significant (p<0.001). There were no statistically significant effects of laser photocoagulation on plasma VEGF, Ang-2, and tie-2 in the 19 patients with PDR, or any effects of eprosartan in the 28 patients with non-proliferative diabetic retinopathy. CONCLUSION: Increased plasma levels of VEGF and Ang-2, as well as lower soluble tie-2, were found in diabetic patients. The highest VEGF and Ang-2 levels were seen among patients with pre-proliferative and proliferative retinopathy, but there was no relation of tie-2 to the severity of retinopathy. As the majority of previous research into Ang-2 and tie-2 has been in relation to angiogenesis and malignancy, the present study would suggest that Ang-2 and tie-2 may be used as potential indices of angiogenesis in diabetes mellitus (in addition to VEGF) and may help elucidate the role of the angiopoietin/tie-2 system in this condition. PMID- 15548810 TI - Corneal and conjunctival sensitivity to air stimuli. AB - AIMS: To determine the repeatability of ocular surface sensitivity to mechanical stimulation using air stimuli and the effect of contact lens (CL) wear on sensitivity. METHODS: Repeatability: 14 subjects (24-39 years) participated. Mechanical sensitivity to warmed (34 degrees C) and ambient (20 degrees C) air was measured for the central cornea (CC), inferior cornea (IC), and inferior conjunctiva (ICON). Measurements were taken on 12 days; six morning and six afternoon measurements. Differences between sites, time of day, and stimulus temperature were evaluated. CL wear: 10 subjects (22-30 years) participated. Measurements were taken at the same time of day, either following no wear, wear of a CL of oxygen permeability [Dk] of 28 x 10(-9) [cm/s][ml O(2)/ml mm Hg] or wear of a CL of Dk 140 x 10(-9) [cm/s][ml O(2)/ml mm Hg]. Differences between sites and wear conditions were evaluated. RESULTS: Repeatability: Sensitivity varied between sites (p<0.01), time of day (p<0.05), and stimulus temperatures (p<0.01). There were no significant differences between days. Mean thresholds for eye temperature stimuli were; CC 64.4 (SD 28.6) ml/min; IC 84.6 (40.0) ml/min; ICON 120.6 (40.4) ml/min and for ambient temperature stimuli were CC 53.9 (16.0) ml/min, IC 59.0 (20.0) ml/min; ICON 72.6 (43.7) ml/min. CL wear: Sensitivity varied between sites and wear conditions (p<0.05). Conjunctival sensitivity was increased after wear of highly oxygen permeable CLs but unaffected by wear of low oxygen permeable CLs. CONCLUSIONS: The prototype gas aesthesiometer is able to repeatably measure ocular surface sensitivity and measurements are consistent with previously reported techniques. PMID- 15548811 TI - Refractive adaptation in amblyopia: quantification of effect and implications for practice. AB - AIM: To describe the visual response to spectacle correction ("refractive adaptation") for children with unilateral amblyopia as a function of age, type of amblyopia, and category of refractive error. METHOD: Measurement of corrected amblyopic and fellow eye logMAR visual acuity in newly diagnosed children. Measurements repeated at 6 weekly intervals for a total 18 weeks. RESULTS: Data were collected from 65 children of mean (SD) age 5.1 (1.4) years with previously untreated amblyopia and significant refractive error. Amblyopia was associated with anisometropia in 18 (5.5 (1.4) years), strabismus in 16 (4.2 (0.98) years), and mixed in 31 (5.2 (1.5) years) of the study participants. Mean (SD) corrected visual acuity of amblyopic eyes improved significantly (p<0.001) from 0.67 (0.38) to 0.43 (0.37) logMAR: a mean improvement of 0.24 (0.18), range 0.0-0.6 log units. Change in logMAR visual acuity did not significantly differ as a function of amblyopia type (p = 0.29) (anisometropia 0.22 (0.13); mixed 0.18 (0.14); strabismic 0.30 (0.24)) or for age (p = 0.38) ("under 4 years" 0.23 (0.18); "4-6 years" 0.24 (0.20); "over 6 years" 0.16 (0.23)). CONCLUSION: Refractive adaptation is a distinct component of amblyopia treatment. To appropriately evaluate mainstream therapies such as occlusion and penalisation, the beneficial effects of refractive adaptation need to be fully differentiated. A consequence for clinical practice is that children may start occlusion with improved visual acuity, possibly enhancing compliance, and in some cases unnecessary patching will be avoided. PMID- 15548812 TI - Factors influencing visual acuity after intravitreal triamcinolone acetonide as treatment of exudative age related macular degeneration. AB - AIM: To evaluate factors influencing change in visual acuity (VA) after intravitreal injection of triamcinolone acetonide as treatment of exudative age related macular degeneration (AMD). METHODS: This prospective, interventional, comparative non-randomised clinical case series study included 94 patients (99 eyes) showing progressive exudative AMD with occult (n = 61 eyes), minimally classic (n = 18), predominantly classic (n = 1), or totally classic (n = 8) subfoveal neovascularisation. Mean follow up was 8.5 (SD 4.7) months (median, 7.3 months; range 3.1-24.5 months). All patients received an intravitreal injection of 20-25 mg of triamcinolone acetonide. RESULTS: An increase in best VA of at least one line on the Snellen charts was found in 63 (63.1%) eyes. Correspondingly, mean VA increased significantly (p<0.001) from 0.17 (SD 0.13) to 0.22 (SD 0.17) after the injection. Postoperative increase in VA was significantly (p<0.001) and negatively correlated with preoperative VA (correlation coefficient, -0.49). Gain in visual acuity was significantly (p = 0.009) higher if preoperative visual acuity was less than 0.08 (gain: 3.2 (SD 2.9) Snellen lines) than if preoperative VA ranged between 0.08 and 0.20 (gain: 1.2 (SD 2.2) Snellen lines). Change in VA was significantly (p = 0.016) less if preoperative VA was higher than 0.20 (change: -0.8 (SD 3.4) Snellen lines). Maximal gain in VA was significantly (p = 0.035) larger in eyes with retinal pigment epithelium detachment than in eyes with minimally classic subfoveal neovascularisation. This was statistically independent of age (p = 0.99), refractive error (p = 0.88), sex (p = 0.92), and duration of follow up (p = 0.46). CONCLUSIONS: Gain in VA after intravitreal injection of 20-25 mg of triamcinolone acetonide is significantly and negatively correlated with preoperative VA. It is significantly larger in eyes with retinal pigment epithelium detachment than in eyes with minimally classic subfoveal neovascularisation. PMID- 15548813 TI - The relation between c-myc expression and interferon sensitivity in uveal melanoma. AB - BACKGROUND/AIM: Interferons (IFN) are currently being used to treat melanoma, including some patients with uveal melanoma. IFN is thought to inhibit tumour growth through downregulation of the c-myc oncogene; the overexpression of which has been shown to be associated with resistance in cell lines. The aim of this study was to investigate the relation between c-myc gene expression and IFN sensitivity in a series of uveal melanomas in a short term chemosensitivity assay. METHODS: Tumours from 45 patients with uveal melanoma who had undergone enucleation were studied. The ATP chemosensitivity assay was used to study sensitivity to IFN-alpha-2b in freshly isolated cells from each tumour. Flow cytometry was used to assess c-myc expression in formalin fixed material from the primary specimens. RESULTS: There was a wide range of IFN sensitivity between the specimens whereas c-myc expression was universal and present in 80% of the tumour cells in 80% of the specimens. Higher c-myc expression was associated with IFN alpha resistance as measured by the maximum percentage of inhibition (p = 0.05) and there was a trend with the IFN sensitivity index (p = 0.07). CONCLUSIONS: These results demonstrate that tumours with high c-myc expression are also associated with IFN resistance. Future research is required to explore the potential of c-myc gene manipulation combined with IFN therapy. PMID- 15548814 TI - Effect of niacin on the choroidal circulation of patients with age related macular degeneration. AB - AIM: To investigate the effects of niacin on choroidal blood flow in age related macular degeneration (AMD). METHODS: 12 AMD patients with bilateral drusen and visual acuity of 20/40 or better in the study eye received a single oral dose of niacin (six subjects received 500 mg and six received 250 mg) or matching placebo on two separate occasions. Laser Doppler flowmetry was used to assess relative choroidal blood velocity (ChB(Vel)), volume (ChB(Vol)), and flow (ChB(Flow)) in the foveola of the study eye at baseline, 30, and 90 minutes after dosing. RESULTS: In comparison with placebo, a statistically significant 24% increase in ChB(Vol) was observed 30 minutes after niacin administration (ANOVA, p = 0.01). In comparison with placebo, a significant decrease in ChB(Vel) of 23% was observed in the 500 mg group (p = 0.04) and no significant change in ChB(Vel) was seen in the 250 mg group at 30 minutes. No significant changes in ChB(Flow) were detected at 30 or 90 minutes. Also, there were no statistically significant changes in ChB(Vol) or ChB(Vel) at 90 minutes. CONCLUSION: In comparison with placebo, a significant 24% increase in ChB(Vol) was observed 30 minutes after niacin administration. Owing to simultaneous decrease in ChB(Vel), however, no significant change in ChB(Flow) was detected. PMID- 15548815 TI - Acute orbital effects of retrobulbar injection on optic nerve head topography. AB - AIM: To assess the effects of acute orbital volume changes after retrobulbar injection on optic nerve head topography. METHODS: The study population consisted of 95 patients with type 2 diabetes mellitus with clinically significant macular oedema who required focal pattern laser photocoagulation therapy in one eye. Before each laser treatment, 49 patients required a retrobulbar injection (approximately 7 ml of a mix of lidocaine 2% with epinephrine and bupivacaine 0.75% in equal volumes) to provide ocular akinesia. Both eyes of all patients underwent optic nerve head topographic analysis once before laser treatment (within 30 minutes), and repeated within 1 hour, 1 day, 1 week, 2 weeks, and 4 weeks after treatment, respectively. Topographic analyses were performed using a confocal scanning laser ophthalmoscope, HRT-II. The disc area, topography standard deviation, and a total of 12 topographic parameters were calculated by HRT-II. RESULTS: The mean age of the patients was 37.9 (SD 3.2) years. The mean disc area of the subjects was 2.12 (0.44) mm(2). Fellow eyes which were not treated with laser, and those treated eyes which did not receive retrobulbar injection before therapy were found not to reveal significant changes in disc topography in any of the examinations (all p values >0.05). In the topographic examinations in the first hour, first day, and first week, laser treated eyes which underwent retrobulbar injection demonstrated significant increase in the disc area, rim area, rim volume, rim area/disc area, and cup shape measure parameters while optic cup parameters significantly decreased (all p values <0.05). In the second week examinations, they did not show significant difference in disc area measurements (p>0.05). By the fourth week, all of the optic nerve head topographic variables were not significantly different from the pre injection values (all p values >0.05). Colour stereoscopic photographs did not reveal any differences in optic disc appearance. CONCLUSION: Acute orbital volume change following retrobulbar injection may cause significant topographic evidence of optic disc oedema lasting approximately 1 week. Significant changes in optic rim and cup area may last for 2 weeks after injection, with all topographic changes returning to baseline by 1 month after injection. The present findings could be a model to reflect the pathological processes that occur in cases of acute orbital volume changes such as retrobulbar haemorrhage. PMID- 15548816 TI - Age related change of optokinetic nystagmus in healthy subjects: a study from infancy to senescence. AB - BACKGROUND: Optokinetic nystagmus (OKN) gain is asymmetrical between temporal to nasal (TN) and nasal to temporal (NT) stimulation in infancy and decreases at older ages. The age at which OKN gain becomes symmetrical and decreases is debated. The aim was to investigate OKN over the whole lifespan in a large sample of healthy subjects. METHODS: In a prospective, cross sectional study OKN was tested monocularly using TN and NT small field stimulation. Stimulation velocity was 15 degrees /s and 30 degrees /s for children aged under 1 year (n = 97), and 15 degrees /s, 30 degrees /s, 45 degrees /s, and 60 degrees /s for older subjects (1-9 years, n = 66; 10-89 years, n = 86). Gain was measured using infrared oculography. RESULTS: Significant OKN gain asymmetry in favour of TN versus NT stimulation was found during the first 5 months of life (p<0.05). Only at 11 months of age was OKN symmetrical in 100% of the subjects. The percentage of children with symmetrical OKN decreased with increasing stimulus velocity. OKN gain increased in the second and third years (p<0.05 for 15 degrees /s), remained stable until 50 years of age, and showed a small but significant decrease afterwards for the tested velocities (between 6% and 18%, p<0.05). CONCLUSIONS: Infrared oculography is an accurate method to assess OKN, especially in children. Knowledge about change of OKN in healthy subjects could be helpful to interpret OKN in patients with abnormal binocular vision or lesions of the central nervous system. PMID- 15548817 TI - The evaluation of a humidifying device for vitreoretinal surgery. AB - AIM: To study the feasibility of humidifying air during vitreoretinal surgery and measure the water content of air before and after intraocular transit. METHODS: The absolute water content of air was measured in a series of six eyes undergoing fluid-air exchange during macular hole surgery. Infrared absorption spectroscopy was used to determine the water content of the air infusing and exiting each eye. After baseline measurements for each eye were recorded, a second fluid-air exchange was performed and the effect of humidifying the air infusion was documented. The humidifying device used in this study was a prototype adapted from a commercially available respiratory humidifier and enables humidified air to be delivered at a controlled temperature. RESULTS: The water content of air increased following intraocular transit, implying dehydration occurs from the intraocular surfaces. For a standard airline infusion the mean increase in water content of air egressing from an eye was 13.4 mg/l. Humidifying the air reduced the rate of water loss by nearly 90%. CONCLUSIONS: Significant water losses can occur from eyes undergoing fluid-air exchange. Humidifying the infused air can substantially reduce the dehydrating effect during an air exchange. This outcome may have a beneficial effect in reducing cataract formation and visual field defects associated with macular hole surgery. PMID- 15548818 TI - Effects of trypan blue on cell viability and gene expression in human retinal pigment epithelial cells. AB - AIM: To evaluate the effects of trypan blue on cell viability and gene expression in human retinal pigment epithelial (RPE) cells. METHODS: Three concentrations (0.06 mg/ml, 0.6 mg/ml, and 4 mg/ml) of trypan blue were applied to human ARPE19 cells for 1 minute. Cell viability was measured using the 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. RPE cells were sampled daily for 6 consecutive days to assess the effects of trypan blue on cell viability. The effects of trypan blue on the expression of apoptosis related and cell cycle arrest gene expressions including c-fos, c-jun, p53, and p21 were performed using reverse transcription-polymerase chain reaction and immunostaining. RESULTS: The MTT assay showed a concentration dependent suppression effect of trypan blue on cell viability, with higher reduction in the 0.6 mg/ml and 4 mg/ml trypan blue treated groups. No significant change in the expression of c-fos and c-jun was found with all three concentrations of trypan blue. An increase in p53 expression was found in the 4 mg/ml trypan blue treated group at 10-30 minutes after trypan blue application. Immunostaining showed a mild, albeit insignificant, increase of p53 expression in the RPE cells. No significant increase in p21 expression was observed in the 0.06 mg/ml trypan blue treated group but there were significant increases in p21 expression in both the 0.6 mg/ml (p = 0.032) and the 4 mg/ml (p = 0.025) treated groups. CONCLUSIONS: Trypan blue may lead to toxicity on cultured RPE cells as indicated by the reduction in cell viability and changes in the expression of apoptosis related and cell cycle arrest genes at higher concentrations. The application of 0.06 mg/ml trypan blue for 1 minute appeared to have no significant effect on cultured RPE. PMID- 15548819 TI - The use of magnetic resonance imaging in the diagnosis of suspected giant cell arteritis. PMID- 15548820 TI - Bilateral ischaemic optic neuropathy and stroke after multiple bee stings. PMID- 15548821 TI - Cause of V pattern strabismus in craniosynostosis: a case report. PMID- 15548822 TI - West Nile virus chorioretinitis. PMID- 15548823 TI - Swimming goggles suck. PMID- 15548824 TI - Immune recovery disease: a case of interstitial keratitis and tonic pupil following bone marrow transplantation. PMID- 15548825 TI - Occult macular dystrophy in an 11 year old boy. PMID- 15548826 TI - Pseudomonas aeruginosa microbial keratitis secondary to cosmetic coloured contact lens wear. PMID- 15548827 TI - Severe proliferative retinopathy in a patient with advanced muscular dystrophy. PMID- 15548828 TI - Bilateral decompression retinopathy after orbital decompression surgery. PMID- 15548829 TI - Retinal nerve fibre layer damage after indocyanine green assisted vitrectomy. PMID- 15548830 TI - Functional genomics of the dopaminergic system in hypertension. AB - Abnormalities in dopamine production and receptor function have been described in human essential hypertension and rodent models of genetic hypertension. Under normal conditions, D(1)-like receptors (D(1) and D(5)) inhibit sodium transport in the kidney and intestine. However, in the Dahl salt-sensitive and spontaneously hypertensive rats (SHRs) and in humans with essential hypertension, the D(1)-like receptor-mediated inhibition of epithelial sodium transport is impaired because of an uncoupling of the D(1)-like receptor from its G protein/effector complex. The uncoupling is receptor specific, organ selective, nephron-segment specific, precedes the onset of hypertension, and cosegregates with the hypertensive phenotype. The defective transduction of the renal dopaminergic signal is caused by activating variants of G protein-coupled receptor kinase type 4 (GRK4: R65L, A142V, A486V). The GRK4 locus is linked to and GRK4 gene variants are associated with human essential hypertension, especially in salt-sensitive hypertensive subjects. Indeed, the presence of three or more GRK4 variants impairs the natriuretic response to dopaminergic stimulation in humans. In genetically hypertensive rats, renal inhibition of GRK4 expression ameliorates the hypertension. In mice, overexpression of GRK4 variants causes hypertension either with or without salt sensitivity according to the variant. GRK4 gene variants, by preventing the natriuretic function of the dopaminergic system and by allowing the antinatriuretic factors (e.g., angiotensin II type 1 receptor) to predominate, may be responsible for salt sensitivity. Subclasses of hypertension may occur because of additional perturbations caused by variants of other genes, the quantitative interaction of which may vary depending upon the genetic background. PMID- 15548831 TI - Whole blood and leukocyte RNA isolation for gene expression analyses. AB - The analysis of gene expression data in clinical medicine has been plagued by the lack of a critical evaluation of accepted methodologies for the collection, processing, and labeling of RNA. In the present report, the reliability of two commonly used techniques to isolate RNA from whole blood or its leukocyte compartment was compared by examining their reproducibility, variance, and signal to-noise ratios. Whole blood was obtained from healthy subjects and was either untreated or stimulated ex vivo with Staphylococcus enterotoxin B (SEB). Blood samples were also obtained from trauma patients but were not stimulated with SEB ex vivo. Total RNA was isolated from whole blood with the PAXgene proprietary blood collection system or from isolated leukocytes. Biotin-labeled cRNA was hybridized to Affymetrix GeneChips. The Pearson correlation coefficient for gene expression measurements in replicates from healthy subjects with both techniques was excellent, exceeding 0.985. Unsupervised analyses, including hierarchical cluster analysis, however, revealed that the RNA isolation method resulted in greater differences in gene expression than stimulation with SEB or among different trauma patients. The intraclass correlation, a measure of signal-to noise ratio, of the difference between SEB-stimulated and unstimulated blood from healthy subjects was significantly higher in leukocyte-derived samples than in whole blood: 0.75 vs. 0.46 (P = 0.002). At the P < 0.001 level of significance, twice as many probe sets discriminated between SEB-stimulated and unstimulated blood with leukocyte isolation than with PAXgene. The findings suggest that the method of RNA isolation from whole blood is a critical variable in the design of clinical studies using microarray analyses. PMID- 15548832 TI - A model for estimating joint maternal-offspring effects on seed development in autogamous plants. AB - We present a statistical model for testing and estimating the effects of maternal offspring genome interaction on the embryo and endosperm traits during seed development in autogamous plants. Our model is constructed within the context of maximum likelihood implemented with the EM algorithm. Extensive simulations were performed to investigate the statistical properties of our approach. We have successfully identified a quantitative trait locus that exerts a significant maternal-offspring interaction effect on amino acid contents of the endosperm in maize, demonstrating the power of our approach. This approach will be broadly useful in mapping endosperm traits for many agriculturally important crop plants and also make it possible to study the genetic significance of double fertilization in the evolution of higher plants. PMID- 15548834 TI - Eosinophilic leukemia and idiopathic hypereosinophilic syndrome are mutually exclusive diagnoses. PMID- 15548835 TI - THE problem in orthodontics. PMID- 15548836 TI - Use of bonded power arms in cases with missing upper lateral incisors. PMID- 15548837 TI - The Readers' Corner. 1. Do you use indirect bonding? PMID- 15548838 TI - The Readers' Corner. 2. What percentage of your patients are being treated with Invisalign appliances? PMID- 15548839 TI - An acrylic transfer tray for direct-bonded lingual retainers. PMID- 15548840 TI - Direct bonding of Ortho Flextech lingual retainers. PMID- 15548841 TI - A simple transfer tray for bonding lingual retainers. PMID- 15548842 TI - Bonding fiber-reinforced lingual retainers with color-reactivating flowable composite. PMID- 15548843 TI - Orthodontics and implant therapy to replace a congenitally missing lateral incisor. PMID- 15548844 TI - In silico approaches for predicting ADME properties of drugs. AB - Combinatorial chemistry and high-throughput screening have increased the possibility of finding new lead compounds at much shorter time periods than conventional medicinal chemistry. However, too much promising drug candidates often fail because of unsatisfactory ADME properties. In silico ADME studies are expected to reduce the risk of late-stage attrition of drug development and to optimize screening and testing by looking at only the promising compounds. To this end, many in silico approaches for predicting ADME properties of compounds from their chemical structure have been developed, ranging from data-based approaches such as quantitative structure-activity relationship (QSAR), similarity searches, and 3-dimensional QSAR, to structure-based methods such as ligand-protein docking and pharmacophore modelling. In addition, several methods of integrating ADME properties to predict pharmacokinetics at the organ or body level have been studied. In this article, we briefly summarize in silico ADME approaches. PMID- 15548845 TI - Eisai hyperbilirubinemic rat (EHBR) as an animal model affording high drug exposure in toxicity studies on organic anions. AB - The Eisai hyperbilirubinemic rat (EHBR) should be a useful animal model for studies on the toxicity of organic anions which are substrates of multidrug resistance-associated protein 2 (Mrp2), since the systemic exposure to these compounds is expected to be increased in EHBR. In this study, we tested the value of EHBR for this purpose, using pravastatin (PV) and methotrexate (MTX) as model compounds. In the case of a single oral dose of PV (200 mg/kg), C(max) in plasma was 4.0-fold higher and AUC(0-infinity) was 3.6-fold larger than those of normal Sprague-Dawley rats (SDR), respectively. When multiple doses of PV were given to EHBR without co-administration of any other compound, drug-induced skeletal muscle toxicity (myopathy/rhabdomyolysis) and increased creatine phosphokinase (CPK) level were observed, whereas a control experiment using SDR did not show any toxic change. When a single dose of MTX (0.6 mg/kg) was given to EHBR orally, C(max) was 1.7-fold higher and AUC(0-infinity) was 1.6-fold larger than those of SDR, respectively. When multiple doses of MTX were given to EHBR, the changes in bone marrow, spleen and intestines were more severe than those in SDR. These findings support the view that EHBR would be a valuable animal model for toxicity studies on organic anion compounds which are substrates of Mrp2. PMID- 15548846 TI - Utility of a three-dimensional cultured human skin model as a tool to evaluate the simultaneous diffusion and metabolism of ethyl nicotinate in skin. AB - The simultaneous diffusion and metabolism of ethyl nicotinate (EN) in a cultured human skin model, Living Skin Equivalent-high, was evaluated by the in vitro skin permeation and metabolism experiments, and esterase distribution was also determined. Theoretical calculations using Fick's 2nd Law of Diffusion with Michaelis-Menten kinetics were performed to obtain the permeation and metabolic parameters together with information on enzyme distribution. The obtained data was compared with the corresponding results in excised hairless rat skin. The partition coefficient of EN from the vehicle to the skin was of the same order of magnitude for the cultured human skin and hairless rat skin, but the diffusion and metabolic parameters were different. Esterase concentration in the epidermal membrane was greater than in the dermis of cultured skin, which was similar to hairless rat skin. Taking into account the similarities and differences between the membranes, the cultured human skin model can be utilized as a model membrane to rapidly predict simultaneous diffusion and metabolism of the prodrug through human skin. PMID- 15548847 TI - Mechanism of L-lactic acid transport in L6 skeletal muscle cells. AB - L-lactic acid transport plays an important role in the regulation of L-lactic acid circulation into and out of muscle. To clarify the transport mechanism of L lactic acid in skeletal muscle, L-lactic acid uptake was investigated using a L6 cell line. mRNAs of monocarboxylate transporter (MCT) 1, 2 and 4 were found to be expressed in L6 cells. The [(14)C] L-lactic acid uptake by L6 cells increased up to pH of 6.0. The [(14)C] L-lactic acid uptake at pH 6.0 was concentration dependent with a K(m) of 3.7 mM. This process was reduced by alpha-cyano-4 hydroxycinnamate, a typical MCT1, 2 and 4 inhibitor. These results suggest that an MCT participates in the uptake of L-lactic acid by L6 cells. [(14)C] L-lactic acid uptake was markedly inhibited by monocarboxylic acids and monocarboxylate drugs but not by dicarboxylic acids and amino acids. Moreover, benzoic acid, a substrate for MCT1, competitively inhibited this process with K(i) of 1.7 mM. [(14)C] L-lactic acid efflux in L6 cells was inhibited by alpha-cyano-4 hydroxycinnamate but not by benzoic acid. These results suggest that [(14)C] L lactic acid efflux in L6 cells is mediated by MCT other than MCT1. PMID- 15548848 TI - Methotrexate-loxoprofen interaction: involvement of human organic anion transporters hOAT1 and hOAT3. AB - Human organic anion transporters hOAT1 (SLC22A6) and hOAT3 (SLC22A8) are responsible for renal tubular secretion of an antifolic acid methotrexate, and are considered to be involved in drug interaction of methotrexate with nonsteroidal anti-inflammatory drugs (NSAIDs). In our hospital, a delay of methotrexate elimination was experienced in a patient with Hodgkin's disease, who took loxoprofen, a commonly used NSAID in Japan, which suggested a cause. In this study, we examined the drug interaction via hOAT1 and hOAT3, using Xenopus laevis oocytes. hOAT1 and hOAT3 mediated the methotrexate transport with low affinity (K(m) of 724.0 muM) and high affinity (K(m) of 17.2 muM), respectively. Loxoprofen and its trans-OH metabolite, an active major metabolite, markedly inhibited the methotrexate transport by both transporters. Their inhibition concentrations (IC(50)) were in the range of the therapeutic levels. These findings suggest that loxoprofen retards the elimination of methotrexate, at least in part, by inhibiting hOAT1 and hOAT3. PMID- 15548849 TI - Effect of genetic polymorphism of OATP-C (SLCO1B1) on lipid-lowering response to HMG-CoA reductase inhibitors. AB - The effect of genetic polymorphism of human organic anion transporting polypeptide C (OATP-C) on the lipid-lowering response to 3-hydroxy-3 methylglutaryl-CoA (HMG-CoA) reductase inhibitors was assessed. A retrospective study was conducted on 66 patients who underwent treatment of hyperlipidemia with HMG-CoA reductase inhibitors in a municipal hospital in a community-based cohort of Ehime prefecture in the southern part of Japan. Plasma lipid concentrations before and after administration were analyzed in patients in relation to the 521T/C (Val-174-->Ala) polymorphism in the OATP-C gene (TT: n=44 (66.7%), TC: n=20 (30.3%), CC: n=0 (0.0%), undetermined: n=2 (3.0%)). Total cholesterol level was significantly lowered after treatment with HMG-CoA reductase inhibitors in all patients (p<0.001); moreover, subjects with the 521C allele showed an attenuated total-cholesterol-lowering effect compared with those homozygous for the 521T allele (-22.3+/-8.7% vs. -16.5+/-10.5%, p<0.05). These data suggest that the 521T/C polymorphism of the OATP-C gene modulates the lipid-lowering efficacy of HMG-CoA reductase inhibitors. PMID- 15548850 TI - Statistical evaluation of clinical trial design for a population pharmacokinetic study--a case study. AB - A population pharmacokinetic substudy design of a new chemical entity was evaluated based on the bias in parameter estimates and the power of detecting a specific subpopulation showing different clearance using a clinical trial simulation approach. The effect of analysis algorithms on type I error was also assessed. The design factors included the number of patients (n=100-300) and the number of sampling points per patient (n=2-6). Simulation data were generated from a model developed based on a Phase I study. The power was evaluated for a percentile of test statistics obtained by the simulation study. The clearance (CL) related parameters were estimated with sufficient accuracy in all study designs and all analysis algorithms: the first order (FO), first order conditional estimation (FOCE) and first order conditional estimation with interaction (FOCE-INTER) methods. With the FO and FOCE methods, the type I error rate increased as the frequency of sampling from each patient became higher, but such increase was hardly observed with the FOCE-INTER method. The power tended to depend on the size of the subpopulation. A large difference was found in the power of detecting a specific subpopulation showing a clearance decrease of 30% or 50%. Therefore, the most dominant factors controlling power would be the size of the subpopulation and the decreasing ratio of CL in the subpopulation. These findings obtained by the clinical trial simulation approach are useful for optimization of study design and determination of the limits of evaluation. PMID- 15548851 TI - Effect of water intake on pharmacokinetics of lansoprazole from fast disintegrating tablet in human subjects. AB - Lansoprazole fast disintegrating tablet (LFDT) has been developed as a multiple unit formulation to increase the QOL of patients, i.e., easy intake without water. However, there is a possibility that patients intake LFDT in accordance with clarithromycin and amoxicillin with water. To study the effect of water on the absorption of lansoprazole (LPZ), the study was carried out using human volunteers. After selected by phenotype of LPZ metabolism, extensive metabolizers (EMs) of LPZ were used in this study. Twelve healthy male EMs intook LFDT containing 30 mg LPZ with 150 mL of water and without-water, i.e., with saliva, to study the pharmacokinetics of LPZ from the gastrointestinal tract by a cross over manner with one-week washout period under fasted condition in the morning. The mean AUC(0-24s) were 2004.4+/-973.6 ng.h/mL in without-water experiment and 2018.5+/-1159.6 ng.h/mL in the case of with-water experiment. Mean C(maxs) were 851.9+/-450.8 ng/mL in without-water experiment and 830.8+/-456.8 ng/mL in with water experiment, respectively. ANOVA was applied to the log-transformed AUC(0 24) and C(max) values. The 90% two sided confidence intervals for log-transformed AUC(0-24) was 0.78-1.22 and that for log-transformed C(max) was 0.67-1.37, respectively. By comparing these pharmacokinetic parameters, we may state that there was no significant difference between the two administration modes. PMID- 15548852 TI - Molecular mechanisms and drug development in aquaporin water channel diseases: aquaporin superfamily (superaquaporins): expansion of aquaporins restricted to multicellular organisms. AB - Eleven aquaporins are identified in mammals. They all have highly conserved two asparagine-proline-alanine (NPA) boxes that are important for the formation of the water permeating pore. Recently we identified two new proteins in mammals distantly related to aquaporins (AQPs) with poorly conserved NPA boxes. Similarly poorly conserved AQP-like proteins were found in several genome projects of multicellular organisms. They may be subgrouped as the AQP superfamily or superaquaporins. Their function is still unknown. AQP11 knockout mice suffer from polycystic kidneys and neonatal fatality. The identification of superaquaporins will provide new insights into the role of AQPs in organogenesis. PMID- 15548853 TI - Molecular mechanisms and drug development in aquaporin water channel diseases: the translocation of aquaporin-5 from lipid rafts to the apical plasma membranes of parotid glands of normal rats and the impairment of it in diabetic or aged rats. AB - Salivary secretion from rat salivary glands occurs in response to stimulation by acetylcholine and norepinephrine released from nerve endings. Aquaporin-5 (AQP5) localizes in lipid rafts under control conditions and is induced to traffic to the apical plasma membrane in interlobular ducts of rat parotid glands by the activation of M3 muscarinic acetylcholine receptors or alpha1-adrenoceptors. This review will focus on the mechanisms of the translocation of AQP5 from lipid rafts to the apical plasma membrane in the interlobular duct cells of parotid glands of normal rats and the impairment of its translocation in diabetic or senescent rats. PMID- 15548856 TI - Effects of strain on contractile force and number of sarcomeres in series of Xenopus laevis single muscle fibres during long-term culture. AB - The aim of the present study is to test whether mechanical strain uniquely regulates muscle fibre atrophy/hypertrophy and adaptation of the number of sarcomeres in series within mature muscle fibres in vitro . Mature single muscle fibres from Xenopus laevis illiofibularis muscle were cultured (4-97 days) while kept at negative strain ( approximately 20% below passive slack length, 'short fibres') or at positive strain ( approximately 5% over passive slack length, 'long fibres'). Before and after culture the number of sarcomeres in series was determined using laser diffraction. During culture, twitch and tetanic force characteristics were measured every day. Survival time of long fibres was substantially less than that of short fibres. Of the long fibres 40% died or became inexcitable within 1 week, whereas this did not occur for short fibres. During culture, twitch and tetanic force of all short fibres increased substantially. Regression analysis showed that the post-culture number of sarcomeres in series was not significantly changed compared to the number before culture. It is concluded that culture at negative strain does not result in atrophy or a reduction of the number of sarcomeres in series, even after 97 days. For the long fibres we did not detect any hypertrophy as tetanic force remained stable or decreased slowly, while twitch force varied. Regression analysis of the change of the number of sarcomeres in series as a function of the culture time showed a positive slope ( P=0.054). Two out of four long fibres that were cultured for at least 2 weeks showed an increase in the number of sarcomeres of 4 5%. Compared with in vivo adaptation to mechanical stimuli this is much less than would be expected. The data suggest that strain may not be the only factor that regulates hypertrophy and the number of sarcomeres in series. PMID- 15548857 TI - Effect of glucocorticoid receptor ligands on myosin heavy chains expression in rat skeletal muscles during controllable stress. AB - The influence of agonist (dexamethasone) and antagonist (mifepristone) of glucocorticoid receptor during controllable painless stress was evaluated on myosin heavy chains expression in three masticatory and two nape rat muscles: anterior digastric (AD), anterior temporalis (AT), masseter superficialis (MS), longissimus capitis (L) and rectus capitis dorsalis major (R). The relative amounts of myosin heavy chain (MHC) protein isoform contained were significantly affected in four muscles studied by dexamethasone and in three muscles studied under mifepristone, versus control during the stress procedure, after only 1 week of treatment. The control group AT muscles contained respectively 18.2% of MHC 2A, 34.5% of MHC 2X and 47.4% of MHC 2B. The effects of dexamethasone and mifepristone were opposite in this muscle: under dexamethasone, the relative proportions of the three isoforms were 14.2, 31.0 and 54.8%: consequently, MHC 2A and 2X decreased with the profit of 2B. Under mifepristone, the relative proportions were 21.1, 36.6 and 42.3% (MHC 2A and 2X increased to the detriment of 2B). The L muscle was not affected by the two treatments and MS muscle was only affected by dexamethasone. Dexamethasone increased MHC 2B to the detriment of MHC 2A in MS, AD and R. Mifepristone and dexamethasone induced the same changes in AD. The mifepristone treatment decreased the MHC 2X profile in R. Under dexamethasone, four muscles exhibited a significantly higher proportion of the more rapid isoforms than under mifepristone. A previous work showed that controllable stress induced a marked increase in the relative expression of MHC 2B in the same skeletal muscles (Martrette et al. , 1998). Our results confirm then a significant participation of glucocorticoid in MHC isoform expression during controllable stress. PMID- 15548858 TI - Cell migration and metastasis as targets of small RNA-based molecular genetic analyses. AB - Metastatic tumor cells can migrate from one place to another in the body. This involves their adherence to host cell layers and subsequent transcellular movements by a complex process, molecular basis of which are yet to be clarified. Elucidation of genes functionally involved in metastasis may lead to deeper understanding of the mechanism of cell migration, and identification and designing of metastasis-modulating strategies for cancer therapeutics. We review here cell migration in tumor metastasis and the use of small RNA-based approaches to identify functional genes. We then describe our promising novel approach that uses randomized ribozyme libraries for identification of genes involved in cell migration, a consistent feature of metastatic cells. PMID- 15548859 TI - Post-natal developmental expression of alphaKAP splice variants in rabbit fast twitch and slow-twitch skeletal muscle. AB - AlphaKAP is a non-kinase product of a gene within alpha CaMKII gene, that anchors catalytic subunits to skeletal muscle sarcoplasmic reticulum. alphaKAP gene expresses two distinct mRNA transcripts by alternative splicing in rabbit fast muscle, rather than one, as an apparently fiber-type specific and species specific property. We report on the expression profile of alphaKAP gene in fast and slow muscles of adult rats and in rabbit post-natal developing muscles, by RT PCR analysis of muscle RNA, together with Western blot analysis of expressed alphaKAP protein isoforms. We show that alphaKAP isoform predominantly expressed in muscle of newborn rabbits, corresponds to the fast-adult isoform. In fast muscle, the transition from neonatal isoform to the common adult isoform takes place early after birth, and is never complete, unlike in slow muscle. Relevant to these findings, is the observation that neonatal/embryonic alphaKAP mRNA is highly expressed in rat myotubes grown in vitro. Together, our findings suggest that alphaKAP splicing pathways are primarily modulated according to muscle developmental stage. PMID- 15548860 TI - Inactivation of Ca2+ transients in amphibian and mammalian muscle fibres. AB - MagFluo-4 fluorescence (Ca2+) transients associated with action potentials were measured in intact muscle fibres, manually dissected from toads ( Leptodactylus insularis ) or enzymatically dissociated from mice. In toads, the decay phase of the Ca2+ transients is described by a single exponential with a time constant ( tau ) of about 7 ms. In mice, a double exponential function with tau 's of 1.5 and 15.5 ms, respectively gives a better fit. In both species the amplitude of Ca2+ transients diminished during repetitive stimulation: in amphibian muscle fibres, the decrease was about 20% with 1 Hz stimulation and 55% at 10 Hz. In mammalian fibres, repetitive stimulation causes a less conspicuous decrease of the transient amplitude: 10% at 1 Hz and 15% at 10 Hz. During tetanic stimulation at 100 Hz the transient amplitude decays to 20% in toad fibres and 40% in mouse fibres. This decrease could be associated with the phenomenon of inactivation of Ca2+ release, described by other authors. Recovery from inactivation, studied by a double stimuli protocol also indicates that in toad fibres the ability to release Ca2+ is abolished to a greater extent than in mouse fibres. In fact the ratio between the amplitudes of the second and first transient, when they are separated by a 10 ms interval, is 0.29 for toad and 0.58 for mouse fibres. In toad fibres, recovery from inactivation, to about 80 % of the initial value, occurs with a tau of 32 ms at 22 degrees C; while in mouse fibres recovery from inactivation is almost complete and occurs with a tau of 36 ms under the same conditions. The results indicate that Ca2+ release in enzymatically dissociated mammalian muscle fibres inactivates to a smaller extent than in intact amphibian muscle fibres. PMID- 15548861 TI - Intensity of X-ray reflections from skeletal muscle thin filaments partially occupied with myosin heads: effect of cooperative binding. AB - For quantitative analysis of contractile proteins of muscle by means of X-ray diffraction, it is important to know how the intensities of individual reflections are related to the number of diffracting objects, i.e., the amount of constituent contractile protein in the muscle cell. Here we diffused various amounts of exogenous myosin subfragment-1 (S1) into overstretched skinned skeletal muscle fibers, either in the presence or absence of Ca2+ , and derived the relationship between the S1 content and the intensities of reflections arising from the S1. In theory, the intensities should be proportional to the square of the S1 content (square law). However, the intensity-content relation deviated systematically from the square law as the S1 content was lowered, and it was better described as a linear function at the lower end of the S1 contents (<20% of saturation level). Model calculations show that the way of deviation is explained by the cooperative manner of S1 binding to the regulated thin filament. PMID- 15548862 TI - Specificity of blebbistatin, an inhibitor of myosin II. AB - Blebbistatin is a small molecule inhibitor discovered in a screen for inhibitors of nonmuscle myosin IIA. We have examined the specificity and potency of the drug by assaying its effects on the actin-activated MgATPase assay of diverse members of the myosin superfamily. Blebbistatin potently inhibits several striated muscle myosins as well as vertebrate nonmuscle myosin IIA and IIB with IC50 values ranging from 0.5 to 5 microM. Interestingly, smooth muscle which is highly homologous to vertebrate nonmuscle myosin is only poorly inhibited (IC50=80 microM). The drug potently inhibits Dictyostelium myosin II, but poorly inhibits Acanthamoeba myosin II. Blebbistatin did not inhibit representative myosin superfamily members from classes I, V, and X. PMID- 15548863 TI - Neisseria gonorrhoeae porin, P.IB, causes release of ATP from yeast actin. AB - Neisserial porins may play a role in the invasion of the host cell by the bacterium. The protein translocates to the host cell membrane and then to the cytosol during the invasive process, and we have shown it interacts with actin in vitro. Here, we have examined the nucleotide-dependence of the interaction of Neisseria porin, P.IB, with fluorescently labeled yeast G actin. Increasing free ATP between 0 to 0.5 mM retards complex formation between the two proteins. The ATP effect probably results from binding of the nucleotide to actin rather than to porin. Complex formation results in a biphasic release of bound nucleoside triphosphate from actin in the absence of free nucleotide at a rate slower than that of complex formation, but it does not induce hydrolysis of the actin-bound nucleotide. ATP prevents the porin-induced distortion of F-actin structure, and addition of ATP to the complex formed in the absence of free nucleotide induces actin polymerization indicating that P.IB stabilizes nucleotide-free G-actin. Our results suggest that P.IB causes an actin conformation change leading to the production of a polymerization-competent nucleotide-free protein. PMID- 15548864 TI - An open or closed case for the conformation of calponin homology domains on F actin? AB - Calponin homology domains link many different proteins to the surface of actin filaments. However, details of the structural interactions involved and the methods used to determine them are controversial. In the case of the actin binding protein utrophin, for example, several models have been proposed for the binding of utrophin's calponin homology domains to actin. We review and evaluate these models and their supporting data. PMID- 15548865 TI - Passive stiffness of Drosophila IFM myofibrils: a novel, high accuracy measurement method. AB - As the smallest muscle-cell substructure that retains the intact contractile apparatus, the single myofibril is considered the optimal specimen for muscle mechanics, although its small size also poses some technical difficulties. Myofibrils from Drosophila indirect flight muscle (IFM) are particularly difficult to study because their high passive stiffness makes them hard to handle, and too resistant to stretch to produce enough elongation for the accurate measurement of sarcomere length change. In this study, we devised a novel method for accurate stiffness measurement of single relaxed myofibrils using microfabricated cantilevers and phase contrast microscopy. A special experimental protocol was developed to minimize errors, and some data analysis strategies were used to identify and exclude spurious data. Remarkably consistent results were obtained from Drosophila IFM myofibrils. This novel, high accuracy method is potentially an effective tool for detecting small passive stiffness change in muscle mutants. PMID- 15548866 TI - Does phosphate release limit the ATPases of soleus myofibrils? Evidence that (A)M. ADP.Pi states predominate on the cross-bridge cycle. AB - The ATPases (+/-Ca2+) of myofibrils from rabbit soleus (a slow muscle) and psoas (a fast muscle) have different Ea: -Ca2+, 78 and 60 kJ/mol and +Ca2+, 155 and 71 kJ/mol, respectively. At physiological temperatures, the two types of myofibrillar ATPase are very similar and yet the mechanical properties of the muscles are different (Candau et al. (2003) Biophys J 85: 3132-3141). Muscle contraction relies on specific interactions of the different chemical states on the myosin head ATPase pathway with the thin filament. An explanation for the Ea data is that different states populate the pathways of the two types of myofibril because the rate limiting steps are different. Here, we put this to the test by a comparison of the transient kinetics of the initial steps of the ATPases of the two types of myofibril at 4 degrees C. We used two methods: rapid flow quench ('cold ATP chase': titration of active sites, ATP binding kinetics, k(cat); 'Pi burst': ATP cleavage kinetics) and fluorescence stopped-flow (MDCC-phosphate binding protein for free Pi; myofibrillar tryptophan fluorescence for myosin head thin filament detachment and ATP cleavage kinetics). We find that, as with psoas myofibrils, the most populated state on the cross-bridge cycle of soleus myofibrils, whether relaxed or activated, is (A)M.ADP.Pi. We propose a reaction pathway that includes several (A)M.ADP.Pi sub-states that are either 'weak' or 'strong', depending on the mechanical condition. PMID- 15548867 TI - Detubulation abolishes membrane potential stabilization in amphibian skeletal muscle. AB - A recently reported stabilization ('splinting') of the resting membrane potential ( Em) observed in amphibian skeletal muscle fibres despite extracellular hyperosmotic challenge has been attributed to high resting ratios of membrane Cl- to K+ permeability ( P Cl/ P K) combined with elevations of their intracellular Cl- concentrations, [Cl-]i, above electrochemical equilibrium by diuretic sensitive cation-Cl-, Na-Cl (NCC) and/or Na-K-2Cl (NKCC), co-transporter activity. The present experiments localized this co-transporter activity by investigating the effects of established detubulation procedures on Em splinting. They exposed fibres to introduction and subsequent withdrawal of 400 mM extracellular glycerol, high divalent cation concentrations, and cooling. An abolition of tubular access of extracellularly added lissamine rhodamine fluorescence, visualized by confocal microscopy, and of the action potential afterdepolarization together confirmed successful transverse (T-) tubular detachment. Fibre volumes, V , of such detubulated fibres, determined using recently introduced confocal microscope-scanning methods, retained the simple dependence upon 1/[extracellular osmolarity], without significant evidence of the regulatory volume increases described in other cell types, previously established in intact fibres. However detubulation abolished the Em splinting shown by intact fibres. Em thus varied with extracellular osmolarity in detubulated fibres studied in standard, Cl(-)-containing, Ringer solutions and conformed to simple predictions from such changes in assuming that intracellular ion content was conserved and membrane potential change DeltaEm was principally determined by the K+ Nernst potential. Furthermore, cation--Cl- co-transport block brought about by [Cl-]o or [Na+]o deprivation, or inclusion of bumetanide (10 microM) and chlorothiazide (10 microM) in the extracellular fluid gave similar results. When taken together with previous reports of significant Cl- conductances in the surface membrane, these findings suggest a model that contrastingly suggests a T tubular location for cation--Cl- co-transporter activity or its regulation. PMID- 15548868 TI - Detubulation experiments localise delayed rectifier currents to the surface membrane of amphibian skeletal muscle fibres. AB - Ionic currents in intact and detubulated frog sartorius muscle fibres were compared at room temperature using a loose-patch voltage clamp configuration in four experimental groups. The test fibres (i) were detubulated by a previously established osmotic shock protocol that involved the introduction and withdrawal of extracellular glycerol followed by exposure to Ca2+/Mg2+-Ringer solution and cooling. The control fibres were spared osmotic shock and (ii) simply studied in normal Ringer solution, (iii) exposed to 30 min of steady cooling to 9-10 degrees C before electrophysiological study or (iv) exposed to and studied in glycerol Ringer solution. The presence or absence of detubulation was confirmed for all the experimental groups through assessing for the abolition or otherwise of the delayed after-depolarisation normally associated with action potential propagation into the transverse (T) tubules. All fibre groups showed similar resting potentials (-80 to -90 mV) thus ensuring consistent baseline voltages from which the voltage clamp steps were imposed. The intact muscle fibres in the three control groups (ii)-(iv) spared osmotic shock showed both inward Na+ and delayed rectifier outward (K+) currents. In contrast, patches from detubulated muscle fibres in the test group (i) showed only delayed outward currents, consistent with contrasting contributions to Na+ and K+ currents from regions of membrane affected or spared by the detubulation procedure. Nevertheless, the voltage dependence, maximum steady state amplitudes and timecourses of the delayed outward currents were conserved through all the experimental groups. These findings suggest that the surface as opposed to the tubular membrane contributes the greater part of the delayed rectifier current in amphibian skeletal muscle. PMID- 15548869 TI - Creatine kinase is an alpha myosin heavy chain 3'UTR mRNA binding protein. AB - Altered cardiac workload regulates the translation and localization of the alpha myosin heavy chain (alphaMyHC) messenger RNA through the 3' untranslated region (UTR) by protein-RNA interactions. We used the alphaMyHC 3'UTR from neonatal rat heart tissue in a gel shift analysis to find RNA binding proteins. One was identified by microsequencing as creatine kinase, brain form B (CKBB). The affinity of its binding interaction was evaluated using sense and antisense alphaMyHC 3'UTR and 3'UTR probes from myosin isoforms of 2B and 2X skeletal muscle. Removal of calcium by the chelating agent EGTA had a potentiating effect on the formation of the CKBB/alphaMyHC 3'UTR complex in vitro . Varying the concentration of ATP (0.1-1 mM) also enhanced this interaction, suggesting that autophosphorylation of CKBB is taking place. Our novel finding that CKBB, an energy transduction enzyme, binds to the RNA of the 3'UTR of the faster ATP consuming alphaMyHC suggests a possible regulatory linkage between the metabolic state of the cell and myosin isoform expression. PMID- 15548870 TI - Age-related changes in glucose utilization and fatty acid oxidation in a muscle specific manner during rabbit growth. AB - The optimal utilization of energy substrates in muscle fibers is of primary importance for muscle contraction and whole body physiology. This study aimed to investigate the age-related changes in some indicators of glucose catabolism and fatty acid oxidation in muscles of growing rabbits. Longissimus lumborum (fast twitch, LL) and semimembranosus proprius (slow-twitch, SMP) muscles were collected at 10 or 20 weeks of age ( n=6 per age). Glucose transporter GLUT4 content was investigated by immunoblot assay. Activity levels of five enzymes were measured: lactate dehydrogenase (LDH) and phosphofructokinase (PFK) for glycolysis; citrate synthase (CS), isocitrate dehydrogenase (ICDH) and -3 hydroxyacyl-coenzyme A dehydrogenase (HAD) for oxidation. Mitochondrial and peroxisomal oxidation rates were assessed on fresh homogenates using [1-14C] oleate as substrate. At both ages, mitochondrial and peroxisomal oxidations rates, as well as activities of oxidative enzymes were higher in SMP than in LL. In both muscles, the apparent rate of fatty acid oxidation by the mitochondria did not differ between the two ages. However, a decrease in the activities of the three oxidative enzymes was observed in LL, whereas activities of CS and HAD and peroxisomal oxidation rate of oleate increased between the two ages in SMP muscle. In both muscles, LDH activity increased between 10 and 20 weeks, without variations in glucose uptake (GLUT4 transporter content) and in the first step of glucose utilization (PFK activity). In conclusion, mitochondrial oxidation rate of fatty acids and activities of selected mitochondrial enzymes were largely unrelated. Moreover, regulation of energy metabolism with advancing age differed between muscle types. PMID- 15548872 TI - MusLABEL: a program to model striated muscle A-band lattices, to explore crossbridge interaction geometries and to simulate muscle diffraction patterns. AB - The program MusLABEL has been devised as a simple aid both in understanding the origin and appearance of fibre diffraction patterns from helical structures and also to simulate the structure and some features of the diffraction patterns from striated muscles and their filament components. Helices are common as preferred conformations in both natural and synthetic macromolecules (e.g. DNA, alpha helices, polysaccharides, synthetic polymers), and they also occur frequently in extended macromolecular aggregates (e.g. actin filaments, myosin filaments, microtubules, amyloid filaments etc). For this reason, a simple way of visualising the kinds of diffraction patterns that these filament structures can give, particularly for the actin and myosin filaments in muscle, can have educational value and can also be useful as a quick means of evaluating possible symmetries in structural interpretations of diffraction data before embarking on full helical diffraction analysis. A feature of the MusLABEL program is that, when a particular kind of A-band lattice has been set up, for example for vertebrate striated muscle or insect flight muscle, additional parameters can be defined both to describe the limits to the azimuthal and axial ranges over which a myosin head can search for an actin binding site and also to describe the size and position of an actin 'target area' assuming that the azimuthal position of an actin monomer has a large effect in determining whether or not a myosin head can bind to it. By this means the effects of lattice geometry on head attachment can be explored and the diffraction effects of specific labelling patterns on actin can be calculated and simulated. The MusLABEL program, running under Microsoft Windows, is available free on the CCP13 website (www.ccp13.ac.uk) where further documentation is given. PMID- 15548871 TI - Heme oxygenase-2 products activate IKCa: role of CO and iron in guinea pig portal vein smooth muscle cells. AB - Hemin (10 microM) and carbon monoxide (CO) increased iberiotoxin-blockable IKCa in portal vein smooth muscle cells. CO-induced IKCa activation was abolished by 10 microM ODQ, 10 microM cyclopiazonic acid and 1 microM KT5823. The hemin induced effect on IKCa was abolished by pretreatment with Sn-protoporphyrin IX, a heme oxygenase inhibitor and Fe2+ chelator but was insensitive to inhibitors of soluble guanylate cyclase (GC) and cGMP-dependent protein kinase (PKG). There was no effect of hemin on IKCa in the presence of 3 microM dithiotreitol into the bath or 3 mM glutathione into the pipette solution. Superoxide dismutase (1000 U/ml) or catalase (3000 U/ml) added into the pipette solution also abolished the effect of hemin on IKCa in this tissue. Additionally, 10 microM hemin could not influence IKCa in Ca2+-free external solution or in the presence of 30 microM SKF 95356. It was concluded that CO increases IKCa via its "conventional" signaling pathway, which involves soluble GC and PKG activation and subsequent stimulation of sarcoplasmic reticulum Ca2+ pump activity resulting in Ca2+-dependent activation of IKCa due to the accumulation of Ca2+ into the space near the plasma membrane. On the other hand, internally produced CO could not yield the same IKCa increase, while Fe2+ derived from heme oxygenase 2-dependent degradation of hemin in portal vein smooth muscle cells gives rise to reactive oxygen species namely hydroxyl and superoxide radicals. Both radicals are responsible for the SKF 95356 sensitive non-selective cation channel activation, the Ca2+ influx and the subsequent increase of Ca2+ concentration near the plasma membrane that augments the KCa channel activity. PMID- 15548873 TI - Nanoporous SiC: a candidate semi-permeable material for biomedical applications. AB - We have fabricated free-standing SiC nanoporous membranes in both p -type and n type material. We showed that these membranes will permit the diffusion of proteins up to 29000 Daltons, while excluding larger proteins. By using radioactively labeled albumin, we also show that porous SiC has very low protein adsorption, comparable to the best commercially available polymer nanoporous membrane. PMID- 15548874 TI - Endothelialized networks with a vascular geometry in microfabricated poly(dimethyl siloxane). AB - One key challenge in regenerating vital organs is the survival of transplanted cells. To meet their metabolic requirements, transport by diffusion is insufficient, and a convective pathway, i.e., a vasculature, is required. Our laboratory pioneered the concept of engineering a vasculature using microfabrication in silicon and Pyrex. Here we report the extension of this concept and the development of a methodology to create an endothelialized network with a vascular geometry in a biocompatible polymer, poly(dimethyl siloxane) (PDMS). High-resolution PDMS templates were produced by replica-molding from micromachined silicon wafers. Closed channels were formed by bonding the patterned PDMS templates to flat PDMS sheets using an oxygen plasma. Human microvascular endothelial cells (HMEC-1) were cultured for 2 weeks in PDMS networks under dynamic flow. The HMEC-1 cells proliferated well in these confined geometries (channel widths ranging from 35 mum to 5 mm) and became confluent after four days. The HMEC-1 cells lined the channels as a monolayer and expressed markers for CD31 and von Willebrand factor (vWF). These results demonstrate that endothelial cells can be cultured in confined geometries, which is an important step towards developing an in vitro vasculature for tissue-engineered organs. PMID- 15548876 TI - On the design and optimization of micro-fluidic dielectrophoretic devices: a dynamic simulation study. AB - Microfabricated interdigitated electrode array is a convenient form of electrode geometry for dielectrophoretic trapping of biological particles within micro fluidic biochips. We have previously reported experimental results and finite element modeling of the holding forces for both positive and negative dielectrophoretic traps on microfabricated interdigitated electrodes within a microfluidic biochip fabricated in silicon with a 12 microm deep chamber and anodic-bonded glass cover. Based on these prior studies, we present in this paper a dynamic study to investigate the stopping capability of dielectrophoretic devices with limited electrode teeth. Simulation results on the issues of design and optimization of the dielectrophoretic devices are also presented and discussed in detail. Simulation results show that the maximum particle stopping distance in a specific device is very sensitive to the chamber height due to the near-electrode nature of DEP force. The relationship between maximum stopping distance and the applied voltage is presented, and the electrode spacing is found to be important in designing the electrode geometry. The spacing should be no less than the chamber height in order to efficiently capture the particles in a relatively short range at a given applied voltage and flow rate. PMID- 15548875 TI - Membrane-based PDMS microbioreactor for perfused 3D primary rat hepatocyte cultures. AB - To improve primary adult rat hepatocyte cultures, two types of PDMS microbioreactors containing a membrane, used as a scaffold for the attachment of cells, were built: one with a commercially-available polyester membrane, the other with a PDMS membrane (5 x 5 microm hole sizes) made in our laboratory. In that way, the relative surface area between blood perfusion and hepatocytes seen in vivo was mimicked and as cells were bathed in both sides by the culture medium they expressed much higher functions. A new technique to build such PDMS membrane was created. With this technique we could build various PDMS membranes with down to 5 x 5 microm holes and with thickness even below 20 microm. During the fifteen days of perfusion in these microbioreactors, good cell attachment then cell reorganization was observed. Moreover compared to static cultures in tissue culture-treated dishes and in cultures in inserts with the same polyester membranes, seven and two times increases in the albumin secretion by the cultured primary rat adult hepatocyte were demonstrated, respectively. Ammonium removal also increased 7 times in perfused cultures compared to static cultures. These new microbioreactors, which closely mimic the in vivo liver architecture, revealed themselves to be very promising tools towards future applications in drug screening or liver tissue engineering. PMID- 15548877 TI - Two-dimensional chemotherapy simulations demonstrate fundamental transport and tumor response limitations involving nanoparticles. AB - Zheng et al. (2004) developed a multiscale, two-dimensional tumor simulator with the capability of showing tumoral lesion progression through the stages of diffusion-limited dormancy, neo-vascularization (angiogenesis) and consequent rapid growth and tissue invasion. In this paper we extend their simulator to describe delivery of chemotherapeutic drugs to a highly perfused tumoral lesion and the tumor cells' response to the therapy. We perform 2-D simulations based on a self-consistent parameter estimation that demonstrate fundamental convective and diffusive transport limitations in delivering anticancer drug into tumors, whether this delivery is via free drug administration (e.g., intravenous drip), or via 100 nm nanoparticles injected into the bloodstream, extravasating and releasing the drug that then diffuses into the tumoral tissue, or via smaller 1 10 nm nanoparticles that are capable of diffusing directly and targeting the individual tumor cell. Even in a best-case scenario involving: constant ("smart") drug release from the nanoparticles; a homogenous tumor of one cell type, which is drug-sensitive and does not develop resistance; targeted nanoparticle delivery, with resulting low host tissue toxicity; and for model parameters calibrated to ensure sufficient drug or nanoparticle blood concentration to rapidly kill all cells in vitro ; our analysis shows that fundamental transport limitations are severe and that drug levels inside the tumor are far less than in vitro , leaving large parts of the tumor with inadequate drug concentration. A comparison of cell death rates predicted by our simulations reveals that the in vivo rate of tumor shrinkage is several orders of magnitude less than in vitro for equal chemotherapeutic carrier concentrations in the blood serum and in vitro, and after some shrinkage the tumor may achieve a new mass equilibrium far above detectable levels. We also demonstrate that adjuvant anti-angiogenic therapy "normalizing" the vasculature may ameliorate transport limitations, although leading to unwanted tumor fragmentation. Finally, our results suggest that small nanoparticles equipped with active transport mechanisms (e.g., chemotaxis) would overcome the predicted limitations and result in improved tumor response. PMID- 15548878 TI - Negative dielectrophoretic force assisted construction of ordered neuronal networks on cell positioning bioelectronic chips. AB - Developing new methods and technologies in order to pattern neurons into regular networks is of utmost scientific interest in the field of neurological research. An efficient method here is developed for trapping neurons and constructing ordered neuronal networks on bioelectronic chips by using arrayed negative dielectrophoretic (DEP) forces. A special bioelectronic chip with well defined positioning electrode arrays was designed and fabricated on silicon substrate. When a high frequency AC signal was applied, the cell positioning bioelectronic chip (CPBC) is able to provide a well-defined non-uniform electric field, and thus generate negative DEP forces. The parameters, such as size of positioning electrode, conductivity of working solution, amplitude and frequency of power signal and cell concentration, were investigated to optimize the performance of the CPBC. When the neuron suspension was added onto the energized bioelectronic chip, the neurons were immediately trapped and quickly formed the predetermined pattern. Neurons may adhere and then be cultured directly on the CPBC, and show good neuron viability and neurite development. The formation of the ordered neuronal networks after two-week culture demonstrates that negative dielectrophoretic force assisted construction of ordered neuronal networks is effective, and it could be used to assist in monitoring functional activities of neuronal networks. PMID- 15548879 TI - NanoLiterBioReactor: long-term mammalian cell culture at nanofabricated scale. AB - There is a need for microminiaturized cell-culture environments, i.e. NanoLiter BioReactors (NBRs), for growing and maintaining populations of up to several hundred cultured mammalian cells in volumes three orders of magnitude smaller than those contained in standard multi-well screening plates. These devices would enable the development of a new class of miniature, automated cell-based bioanalysis arrays for monitoring the immediate environment of multiple cell lines and assessing the effects of drug or toxin exposure. We fabricated NBR prototypes, each of which incorporates a culture chamber, inlet and outlet ports, and connecting microfluidic conduits. The fluidic components were molded in polydimethylsiloxane (PDMS) using soft-lithography techniques, and sealed via plasma activation against a glass slide, which served as the primary culture substrate in the NBR. The input and outlet ports were punched into the PDMS block, and enabled the supply and withdrawal of culture medium into/from the culture chamber (10-100 nL volume), as well as cell seeding. Because of the intrinsically high oxygen permeability of the PDMS material, no additional CO(2)/air supply was necessary. The developmental process for the NBR typically employed several iterations of the following steps: Conceptual design, mask generation, photolithography, soft lithography, and proof-of-concept culture assay. We have arrived at several intermediate designs. One is termed "circular NBR with a central post (CP-NBR)," another, "perfusion (grid) NBR (PG-NBR)," and a third version, "multitrap (cage) NBR (MT-NBR)," the last two providing total cell retention. Three cells lines were tested in detail: a fibroblast cell line, CHO cells, and hepatocytes. Prior to the culturing trials, extensive biocompatibility tests were performed on all materials to be employed in the NBR design. To delineate the effect of cell seeding density on cell viability and survival, we conducted separate plating experiments using standard culture protocols in well-plate dishes. In both experiments, PicoGreen assays were used to evaluate the extent of cell growth achieved in 1-5 days following the seeding. Low seeding densities resulted in the absence of cell proliferation for some cell lines because of the deficiency of cell-cell and extracellular matrix (ECM)-cell contacts. High viabilities were achieved in all designs. We conclude that an instrumented microfluidics-based NanoBioReactor (NBR) will represent a dramatic departure from the standard culture environment. The employment of NBRs for mammalian cell culture opens a new paradigm of cell biology, so far largely neglected in the literature. PMID- 15548880 TI - Electrowetting (EW)-based valve combined with hydrophilic teflon microfluidic guidance in controlling continuous fluid flow. AB - Electrowetting (EW)-based techniques have been widely used in manipulating discrete liquid. However, few articles discussed the controlling of continuous fluid flow by using EW-based techniques. In this paper, an EW-based valve combined with plasma-modified Teflon surface, which serves as a microfluidic guidance, in controlling continuous fluid flow has been demonstrated. The plasma modified Teflon surface is firstly demonstrated for confining continuous fluid flow. The EW-based microfluidic device possesses the functions of a valve and a microchannel without complex moving parts and grooved microchannels. The quantitative characteristics of the EW-based valve are also studied. Propylene carbonate (PC) is firstly demonstrated as the working liquid in the EW-based device because of its applications in parallel oligonucleotide synthesis. It is found that lower valve actuation voltage reduces the deterioration of the valve and improves the valve stability. PMID- 15548883 TI - Cardiac memory: do the heart and the brain remember the same? PMID- 15548884 TI - Ventricular mapping during atrial and right ventricular pacing: relation of electrogram parameters to ventricular tachycardia reentry circuits after myocardial infarction. AB - INTRODUCTION: Ventricular tachycardia (VT) late after myocardial infarction is usually due to reentry in the border zone of the infarct area. Identification of critical parts of the VT reentry circuit by catheter mapping without needing to induce VT is a desirable goal for VT ablation. The aim of this study was to develop a model to predict reentry circuit locations based on characteristics of sinus or paced electrograms and pace mapping (PM) recorded from the infarct region. METHODS: Left ventricular electroanatomic mapping with the CARTO mapping system was performed in 16 male patients with recurrent VT late after myocardial infarction. A total of 1072 left ventricular sites were recorded during atrial pacing (AP) and right ventricular pacing (RVP), and the corresponding electrograms were analyzed for their local activation time (LAT), onset (ONS), end (END), duration (DUR), and amplitude (AMP) in each pacing sequence. At 1041 of these sites, PM was performed; the resulting stimulus to QRS intervals (S-QRS) was determined at 931 sites, the remaining 110 sites did not capture. All the obtained parameters were compared with the location of 18 ablation target areas with a radius of 2 cm defined by success of radiofrequency (RF) ablation or entrainment during VT, or both. RESULTS: Of 1072 sites, 227 (21%) were in the target and 845 (79%) were outside the target. All parameters were significantly different (p < 0.05) in AP and in RVP between inside and outside the target in a univariate analysis. In a multivariate analysis LAT, END, DUR, and AMP in AP, END and AMP in RVP, and S-QRS were independent predictors for the target (p < 0.05). A combination of selected parameters of these predictors (DUR in AP, AMP in RVP, and S-QRS) had a specificity of 64% with a sensitivity of 80% for the target. CONCLUSION: The observations suggest that ablation guided by a combination of abnormal electrograms in different rhythms can be useful to ablate VT and reduce the necessity of VT induction. Anatomically fixed regions of block may be important for reentry and be identifiable during sinus rhythm. PMID- 15548885 TI - Outcome of right ventricular bifocal pacing in patients with permanent atrial fibrillation and severe dilated cardiomiopathy due to Chagas disease: three years of follow-up. AB - OBJECTIVES OF STUDY: Several studies have shown that heart failure may benefit from cardiac resynchronization therapy (CRT). Studies have demonstrated a beneficial effect of right ventricular (RV) bifocal pacing, using two leads at different positions, in similar patient populations. The aim was to evaluate this approach in Chagas disease patients who developed both severe dilated cardiomiopathy and chronic atrial fibrillation. METHODS: The study included 30 patients with a mean age of 52 +/- 6 years (16 male), who had atrioventricular block at functional class II or IV (NYHA). Patients underwent endocardial dual chamber pacemaker implantation with two RV leads-one placed near the RV outflow tract and the other in the apex. Patients were examined by echocardiography, 24 hour Holter, and New York Heart Association (NYHA) class determination before and 3, 6, 12, 18, 24, and 36 months after CRT. RESULTS: Compared to the baseline, the left ventricular ejection fraction increased in the first month of CRT, the left ventricular end diastolic diameter decreased, all patients were downgraded to NYHA class I or II, and the incidence of ventricular arrhythmias decreased. However, these could not be maintained and worsened after 6 months CRT. There was a mortality rate of 43.3% during the first year, and only 23.3% of patients remained alive after 3 years. They underwent an electrophysiological study, which revealed complex arrhythmias justifying implantable cardioverter defibrillator (ICD) in six out of seven patients. CONCLUSION: The favorable effects of RV bifocal pacing could not be maintained beyond the first 6 months, likely due to the ventricular arrhythmias. Therefore, CRT combined with ICD from the outset may be recommended for this patient group. PMID- 15548887 TI - Radiofrequency ablation of ventricular fibrillation and multiple right and left atrial tachycardia in a patient with Brugada syndrome. AB - Brugada syndrome is a well-known form of idiopathic ventricular fibrillation (VF). Few data suggest that this arrhythmia may be triggered by ventricular premature beats (VPBs), and an association with other arrhythmia such as monomorphic ventricular tachycardia (VT) or supraventricular tachycardia (SVT) has been reported. In a highly symptomatic 18-year-old-male patient with this syndrome, frequent episodes of VF, fast polymorphic VT, and fast monomorphic sustained regular tachycardia were observed. The tachycardia episodes were classified as VT or VF and as a consequence received appropriate therapies with the implanted cardioverter defibrillator (ICD). Precipitating VPBs that were stored in the ICD memory and on the electrocardiogram (ECG) exhibited the same morphology as frequent isolated VPBs. During the electrophysiological study, right and left atrial tachycardia (AT) with one-to-one atrioventricular conduction were also induced and successfully ablated. VF was ablated using the same noncontact mapping (NCM) system triggering VPBs from right ventricular outflow tract (RVOT). PMID- 15548886 TI - Impact of chronic kidney disease upon survival among implantable cardioverter defibrillator recipients. AB - BACKGROUND: Chronic kidney disease (CKD) has been linked to serious arrhythmias. We studied the impact of CKD upon implantable cardioverter-defibrillator (ICD) recipients. METHODS AND RESULTS: Baseline estimated glomerular filtration rate (eGFR) was calculated from variables at the time of ICD implantation in 95 patients. Patients with eGFR below 60 ml/min and those with end-stage renal disease (ESRD) were considered to have significant CKD. Among 95 patients who underwent ICD implantation for VT/VF, the mean age was 66.5+/-12.2 years, 27 (29.0%) were women and 20 (21.5%) were African American. The CKD groups (eGFR < 60 ml/min and ESRD) and control group (eGFR > or = 60 ml/min) were similar with respect to background histories and medications. A significant difference in all cause mortality in those with eGFR >or = 60 ml/min, 3 patients (8.6%), compared to either those with eGFR < 60 ml/min, 28 (60.9%), or ESRD 7 patients (58.3%), p < 0.0001, was noted. Proportionately more patients died from arrhythmic deaths in those with eGFR < 60 ml/min, 8 patients (17.39%) and ESRD 3 patients (25%), than those with eGFR > or = 60 ml/min, no patient. P < or = 0.0001. There was progressive increase in DFT's with worsening renal failure. The Cox proportional hazards model for time until death, found independent predictors to be: age, OR = 1.04 (per year), 95% CI 1.00-1.08, p = 0.04; CKD group, OR 2.59, 95% CI 1.27 5.30, p = 0.009; and use of beta-blockers, OR 0.25, 95% CI 0.10-0.61, p = 0.002. CONCLUSIONS: Significant CKD was related to overall poor survival, arrhythmic death and high DFTs. PMID- 15548888 TI - Successful percutaneous ablation of ventricular tachycardia in congenitally corrected transposition of the great arteries, a case report. AB - Ventricular tachycardia (VT) is rarely recognized in patients with congenitally corrected transposition of the great arteries (CCTGA). We describe a 48-year-old woman with CCTGA, systemic atrioventricular valve replacement for Ebsteinoid malformation of the valve, ventricular dysfunction and ventricular tachycardia related to the previous surgical scar. The patient had successful non-contact mapping and radiofrequency ablation of the offending tachycardia substrate. PMID- 15548889 TI - Radiofrequency catheter ablation of idiopathic left ventricular tachycardia originating in both left posterior and anterior fascicles. AB - A 45-year-old woman underwent radiofrequency ablation (RFA) for symptomatic idiopathic left ventricular tachycardia (ILVT). The clinical arrhythmias had two different patterns, a wide QRS tachycardia with right bundle branch block (RBBB) and left axis deviation (LAD) and another with RBBB and right axis deviation (RAD). The electrophysiology study localized the origin of tachycardias to the midinferior and superior ventricular septum, respectively. RFA terminated successfully ILVT with RBBB and LAD morphology, but another pattern could not be ablated. Noncontact mapping revealed the earliest site of activation at the superior septum. RFA at this site terminated successfully ILVT with RBBB and RAD. PMID- 15548890 TI - Carl Friedrich Wilhelm Ludwig (1816-1895). PMID- 15548891 TI - Redox biology of blood. PMID- 15548892 TI - Effects of cell-free hemoglobin on hypoxia-inducible factor (HIF-1alpha) and heme oxygenase (HO-1) expressions in endothelial cells subjected to hypoxia. AB - We have investigated the impact of diaspirin cross-linked hemoglobin (DBBF-Hb), a blood substitute, on cell signaling pathways that are modulated in part by biological peroxides (i.e., hydrogen peroxide, lipid peroxide, and peroxynitrite). Bovine aortic endothelial cells (BAECs) subjected to hypoxia expressed hypoxia-inducible factor (HIF-1alpha) in a time course that paralleled the expressions of heme oxygenase (HO-1). Co-incubation of the oxy form (HbFe(2+)) with hypoxic BAECs resulted in an increase in the expression of HIF 1alpha in a manner that corresponded linearly with the decay of HbFe(2+) and accumulation of the ferric form (HbFe(3+)). Inclusion of HbFe(3+) with hypoxic BAECs produced twice as much expression in the HIF-1alpha and HO-1 proteins as opposed to HbFe(2+) alone, or HbFe(2+) plus hypoxia. In addition, higher and more persistent levels of the ferryl form (HbFe(4+)), due to the consumption of endogenous peroxides, were found in the hypoxic media containing hemoglobin. Nitric oxide (NO) released from an NO donor reduced the levels of HIF-1alpha in the hypoxic cells treated with either HbFe(2+) or HbFe(3+), but had little or no effect on the levels of HO-1. DBBF-Hb modulates key cell-signaling pathways by competing with peroxides required for the deactivation of HIF-1alpha, which may modulate important physiological mediators. PMID- 15548893 TI - The radical and redox chemistry of myoglobin and hemoglobin: from in vitro studies to human pathology. AB - Recent research has shown that myoglobin and hemoglobin play important roles in the pathology of certain disease states, such as renal dysfunction following rhabdomyolysis and vasospasm following subarachnoid hemorrhages. These pathologies are linked to the interaction of peroxides with heme proteins to initiate oxidative reactions, including generation of powerful vasoactive molecules (the isoprostanes) from free and membrane- bound lipids. This review focuses on the peroxide-induced formation of radicals, their assignment to specific protein residues, and the pseudoperoxidase and prooxidant activities of the heme proteins. The discovery of heme to protein cross-linked forms of myoglobin and hemoglobin in vivo, definitive markers of the participation of these heme proteins in oxidative reactions, and the recent results from heme oxygenase knockout/knockin animal model studies, indicate that higher oxidation states (ferryl) of heme proteins and their associated radicals play a major role in the mechanisms of pathology. PMID- 15548894 TI - Heme degradation by reactive oxygen species. AB - Heme proteins play a major role in various biological functions, such as oxygen sensing, electron transport, signal transduction, and antioxidant defense enzymes. Most of these reactions are carried out by redox reactions of heme iron. As the heme is not recycled, most cells containing heme proteins have the microsomal mixed function oxygenase, heme oxygenase, which enzymatically degrades heme to biliverdin, carbon monoxide, and iron. However, the red cell with the largest pool of heme protein, hemoglobin, contains no heme oxygenase, and enzymatic degradation of the red cell heme occurs only after the senescent red cells are removed by the reticuloendothelial system. Therefore, only nonenzymatic heme degradation initiated when the heme iron undergoes redox reactions in the presence of oxygen-producing reactive oxygen species takes place in the red cell. Unlike enzymatic degradation, which specifically attacks the alpha-methene bridge, reactive oxygen species randomly attack all the carbon methene bridges of the tetrapyrrole rings, producing various pyrrole products in addition to releasing iron. This review focuses on the literature related to nonenzymatic heme degradation with special emphasis on hemoglobin, the dominant red cell heme protein. PMID- 15548895 TI - Critical redox and allosteric aspects of nitric oxide interactions with hemoglobin. AB - Nitric oxide (NO) is an important signaling molecule. Relatively long-lived NO adducts at the heme and SH groups of hemoglobin (Hb) could enable NO to carry out long-range signaling functions. In spite of significant advances, there remain as yet unresolved issues regarding the possible role of Hb in moderating NO signaling events that affect blood pressure regulation. In this review, we summarize recent reports concerning the redox and allosteric aspects of NO/Hb interactions that have advanced our understanding of the physiological significance of NO binding to heme groups (forming NO-Hb) and of reactions promoting formation of S-nitrosated Hb (SNO-Hb). Allosteric mechanisms modify the bioactivity of NO/Hb complexes by altering the lifetime of NO-Hb and the properties of SNO-Hb. Redox reactions are significant because of the complex chemistry possible for NO and its oxidation products. Reactions at ferrous and ferric heme sites have differing consequences and affinities for interactions with NO. Moreover, redox changes at heme groups affect reactivity of SH groups and vice versa. In spite of low levels of NO-Hb and SNO-Hb found in vivo, recent findings do not rule out participation of NO-Hb or SNO-Hb in NO-dependent signaling reactions. PMID- 15548896 TI - The red blood cell and vascular function in health and disease. AB - Nitric oxide (NO) is widely accepted as a central regulator of vascular tone and a vast array of other cardiovascular signaling mechanisms. An emerging player in these mechanisms is hemoglobin (Hb), an erythrocytic protein that serves as the archetypical model for an allosteric protein. Specifically, red blood cells (RBC) are suggested to be integral in matching blood flow to tissue oxygen demands. The mechanisms proposed involve the ability of Hb to sense changes in oxygen concentrations and coupling this process to modulating vascular NO levels. The molecular basis of these mechanisms remains under investigation, but is clearly diverse and discussed in this article from the basis of the blood flow responses to hypoxia. Another emerging theme in RBC biology is the role of these cells during inflammatory disease in which disease processes promote the interaction of vascular NO and the RBC. This is exemplified in hemolytic diseases, in which released Hb has drastic affects on vascular homeostasis mechanisms. Additionally, it is becoming evident that RBC express numerous molecules that mediate interactions with the extracellular matrix and cellular mediators of inflammation. The functional implications for such interactions remain unclear but highlight potential roles of the RBC in modulating inflammatory disease. PMID- 15548897 TI - Oxygen sensing in the circulation: "cross talk" between red blood cells and the vasculature. AB - Oxygen (O(2)) sensing in blood and regulation of microvascular tone appear to involve hemoglobin (Hb) conformational changes resulting from O(2) desaturation. This observation has prompted the thought that Hb functions as both an O(2) sensor and regulator of microvasular blood flow to meet local tissue oxygen demand. The mechanism(s) by which this is accomplished has recently been the subject of increasing debate. Three primary hypotheses are described within the literature and include release of adenosine 5'-triphosphate by red blood cells (RBCs), release of S-nitrosylated molecules from RBCs originally bound to beta93 cysteine residues of oxyHb, and nitrite conversion and storage of nitric oxide by Hb at the site of ferric (Fe(3+)) and ferrous (Fe(2+)) Hb. Within extravascular cells, the global regulator of oxygen homeostasis is hypoxia-inducible factor-1 (HIF- 1). This transcriptional factor is tightly regulated by O(2) and cellular redox-sensitive mechanisms. HIF-1 activation is responsible for the up-regulation of proteins, which increase O(2) supply. We believe that there are important and yet unexplored mechanisms by which RBCs can directly or indirectly communicate via redox intermediates with extravascular sites as part of the global O(2) sensing mechanism. PMID- 15548898 TI - Oxygen distribution and respiration by the microcirculation. AB - Longitudinal and radial oxygen gradients in the microcirculation due to oxygen release from arterioles show that in some tissues oxygen is primarily supplied by arterioles and secondarily by capillaries. In several tissues, the arteriolar rate of oxygen exit is too large to be explained by diffusion alone, indicating that in these tissues oxygen consumption of the arteriolar wall in vivo is much greater than that shown in in vitro studies of endothelium and vascular smooth muscle, a phenomenon that may be related to the synthesis autocoids by the endothelium in vivo. The functional significance of the high metabolic rate of the arteriolar vessels may be related to the need of providing a metabolic barrier for protecting the parenchymal tissue from high oxygen levels in arterial blood, thus reducing formation of oxygen free radicals in the perivascular tissue, a supposition supported by the finding that the radial oxygen gradient at the microvascular wall and therefore its rate of oxygen consumption are proportional to local blood oxygen partial pressure (pO(2)). Oxygen consumption by the endothelium and/or smooth muscle is also a factor in causing terminal lymphatic pO(2) to have the lowest oxygen level in the tissue, rendering this compartment most vulnerable in hypoxic conditions. PMID- 15548899 TI - Blood substitutes and redox responses in the microcirculation. AB - Transfusion of hemoglobin-based blood substitutes, designed for their plasma expansion and oxygen transport capabilities, has resulted in some major problems, such as organ dysfunction, during clinical trials. Experimental evidence demonstrates that these hemoglobins damage tissue by producing highly reactive oxygen species. Although cell-free hemoglobin may present a low risk to people with normal redox status, patients who are sick and have a poor antioxidant status may be at risk. Oxidative damage is particularly dangerous in the microcirculation because excess leakage of plasma components into the interstitium will disturb the fluid balance between blood and tissue and alter the kinetics of delivery of intravascularly injected drugs, and endogenous enzymes and hormones, to various tissues. In this review, the redox chemistry of hemoglobin-based blood substitutes is briefly described, and their effects on cultured endothelial cells, and on the exchange properties of the microvasculature, are discussed. Taking into account the possible mechanisms by which oxidative damage can occur, various methods to reduce the deleterious effects of blood substitutes in vivo are evaluated. Finally, several possible cell signaling pathways that are triggered in endothelial cells, in response to modified hemoglobins, are considered in terms of protecting microvascular function. PMID- 15548903 TI - Use of mitomycin C in the pediatric airway. AB - PURPOSE OF REVIEW: Mitomycin C was not used in the field of otolaryngology before 1998. Currently it is commonly used by all pediatric otolaryngologists dealing with airway stenoses. Mitomycin C offers the surgeon a method of modifying healing and diminishing granulation tissue and cicatricial scar. RECENT FINDINGS: There are numerous animal studies showing the beneficial effect of mitomycin C on wound healing in the airway. The human studies rarely have controls and in many cases are case reports. Human studies typically use dosages commonly used in ophthalmology, which are in the low range in relation to the animal airway studies. SUMMARY: This article attempts to summarize the available research on mitomycin C, helping the clinical otolaryngologist choose the proper dosage, indications, and frequency of use. PMID- 15548905 TI - The potential effect of widespread use of pneumococcal conjugate vaccines on the practice of pediatric otolaryngology: the case of acute otitis media. AB - PURPOSE OF REVIEW: This review provides the otolaryngologist with the evolving understanding of various aspects of pneumococcal conjugate vaccines (PCVs) that are related to their effect on the respiratory tract RECENT FINDINGS: The efficacy of PCVs against invasive pneumococcal disease and pneumonia is well established and is documented in several well-conducted studies. However, the effect of PCVs on otitis media is less obvious and more complex. PCVs clearly reduce diseases caused by vaccine-type (VT) pneumococci, but replacement of VT serotypes by non-VT serotypes in nasopharyngeal carriage of Streptococcus pneumoniae is responsible for the increase in acute otitis media (AOM) caused by non-VT serotypes. Furthermore, an increased rate of AOM caused by Haemophilus influenzae and Moraxella catarrhalis was found. Since most antibiotic-resistance in S. pneumoniae is confined to VT serotypes, vaccine use also reduces antibiotic resistance. The reduction of carriage by PCVs is responsible for the reduction of spread of VT pneumococci (herd immunity). Thus a modification of AOM rather than just a simple reduction is seen with the widespread use of PCV. SUMMARY: Acute otitis media in the era of widespread use of PCV is modified. A disease with reduced VT serotypes, reduced antibiotic resistance, and a lower rate of sequelae is to be expected. However, replacement with potential virulent organisms and development of antibiotic resistance in non-VT pneumococci is a possibility that needs careful monitoring. PMID- 15548904 TI - Update on hemangiomas and vascular malformations. AB - PURPOSE OF REVIEW: Our knowledge base in the area of vascular anomalies is growing rapidly. With greater understanding of classification and diagnosis, as well as with the numerous areas of research bringing further awareness on the complexity of these lesions, we are improving our ability to treat them. We will attempt in this article to summarize the developments in the field of vascular anomalies over the last year. RECENT FINDINGS: Emphasis on correct classification is still a high priority in the literature and yet there remains a great deal of misinformation. Many new developments in the basic science of these lesions are allowing better understanding of why these lesions occur while improving our management in these patients. Advances in laser surgery as well as sclerotherapy techniques have improved our ability to treat extensive lesions and also improve patients' quality of life. SUMMARY: Many new and exciting areas of discovery occur almost daily in the field of vascular anomalies. Due to the breadth of this topic, it is certain that not all articles can be reviewed however the author has tried to present the most recent and clinically relevant breakthroughs in the field. PMID- 15548906 TI - Recent advances in the treatment of juvenile angiofibroma. AB - PURPOSE OF REVIEW: Juvenile nasopharyngeal angiofibroma is a rare vascular tumor almost exclusive to the nasopharynx of adolescent males. Traditionally, juvenile nasopharyngeal angiofibroma has been treated surgically using open surgical approaches and has been associated with frustratingly high recurrence rates. This article reviews recent contributions to the study and treatment of this disease. In particular, advances in minimally invasive endoscopic resection of juvenile nasopharyngeal angiofibroma are evaluated. RECENT FINDINGS: The growth patterns of juvenile nasopharyngeal angiofibroma are evaluated. Young age does not appear to correlate with more aggressive disease. The major recent advance in the treatment of juvenile nasopharyngeal angiofibroma has been the application of endoscopic endonasal surgery to the treatment of select tumors. This article reviews the indications and inclusion criteria recently put forth to help select patients for this minimally invasive approach. In properly selected patients with Radkowski stage I and II lesions, recurrence rates range between 0 and 7%. Advanced lesions continue, in most cases, to require open approaches, some of which are also presented. SUMMARY: With proper patient selection, endoscopic resection of juvenile nasopharyngeal angiofibroma is feasible and may be preferable to traditional open approaches. Results suggest that after endonasal resection, disease recurrence is low. Most larger lesions, especially those with intracranial spread, continue to require open approaches for complete resection. PMID- 15548907 TI - Management of lymphatic malformations. AB - PURPOSE OF REVIEW: Innovative otolaryngologists, plastic surgeons, craniofacial surgeons, pediatric surgeons, radiologists, anesthesiologists, neonatologists, obstetricians, and scientists have continued to advance our understanding of the etiology, diagnosis, and treatment of lymphatic malformations. This article reviews the publications over the past 2 years with respect to these advances. RECENT FINDINGS: Fast-sequence MRI limits motion artifacts and allows prenatal MR to be used as a complementary study to ultrasound in the evaluation of large congenital neck masses. Three-dimensional ultrasonography may also be helpful in evaluating prenatal lymphatic malformations. Fluorescence in situ hybridization techniques can be used to evaluate lymphatic malformations for prenatal chromosomal analysis with emphasis on chromosomes 13, 18, 21, X, and Y. The sclerosing agent OK-432 is effective for macrocystic lymphatic malformations but showed less promise for microcystic lesions, mixed lesions, and lesions outside the head and neck region. Somnoplasty shows promise for reduction of tongue lymphatic malformations. Surgical excision, staged when necessary, continues to be integral to management in many cases. SUMMARY: Basic science research has furthered understanding of lymphatic malformations. Clinical research has expanded and refined our diagnostic and therapeutic options for patients with these lesions. Further identification of genes selectively expressed by lymphatic endothelium should facilitate identification of usable vascular markers that can enable analysis of the underlying biology, physiology, pathology, and treatment of the lymphatic system and its malformations. PMID- 15548908 TI - Pediatric chronic rhinosinusitis. AB - PURPOSE OF REVIEW: Pediatric sinusitis is prevalent, and the debate continues regarding how best to care for these children. Although acute sinusitis is commonly associated with an upper respiratory infection, the focus of this paper is on chronic rhinosinusitis in children. Research is often more difficult in children than adults, so many times one can learn from the adult literature and determine whether there can be application to the childhood population. RECENT FINDINGS: This paper looks at both medical and surgical treatment of chronic rhinosinusitis. "Maximal medical management" is often cited in the literature, but what this should consist of has never been clearly proved in the literature. Alternative medicine approaches as well as irrigation as an adjunct to care are discussed. Biomaterials are also be discussed. Recent outcome data are put in perspective. SUMMARY: Hopefully the reader will find the presentation stimulating. The paper does not promote surgery as a "cure all," and in the end, analysis will hopefully leave the reader more cautious but with a better understanding of this complex disease. PMID- 15548909 TI - Management of subglottic hemangioma. AB - PURPOSE OF REVIEW: Subglottic hemangioma is a challenging congenital anomaly involving the larynx in children. Management is not uniform, and no single treatment modality has been accepted as ideal. During the last several years only a few articles have dealt with this topic. The purpose of this review is to examine the published literature and discuss the alternative treatments of SGH. The authors also describe their approach and the use of microdebrider as a new surgical tool. RECENT FINDINGS: Observation is an option for the rare case of SGH with minimal airway obstruction. However, most cases will require some additional intervention. Systemic steroids intralesional steroid injection, laser ablation with both the CO2 and KTP lasers, interferon (IFN) and open surgical excision have all been utilized. SUMMARY: The authors found few technical innovations in this field in the last 5 years. All techniques utilized have some degree of success. However, all approaches have downsides and are associated with complications, some of which can be very serious. Ideally, more structured research comparing techniques would be helpful to best determine operative and postoperative management. PMID- 15548910 TI - Distraction osteogenesis of the mandible. AB - PURPOSE OF REVIEW: Airway obstruction and craniofacial deformities resulting from mandibular deficiency are challenging and complex clinical problems. Mandibular distraction osteogenesis is playing a rapidly expanding role in the treatment of these children and has supplanted traditional management techniques in many centers. Several recent studies have reviewed clinical experience, described technical refinements, and addressed issues in patient selection and preoperative workup. RECENT FINDINGS: The extension of mandibular distraction osteogenesis to newborns with Pierre Robin sequence or craniofacial syndromes associated with micrognathia and airway obstruction has radically altered treatment protocols at several institutions. Early bilateral mandibular distraction has obviated tracheostomy in most newborns, with low operative morbidity. In addition, early decannulation and resolution of obstruction has occurred when mandibular distraction osteogenesis was applied to selected micrognathic children. Some of these patients were tracheotomized and others presented with severe obstructive symptoms. Studies have described success with external and internal distraction techniques. Recent publications have also demonstrated the surgeon's ability to mold the regenerate safely, the dynamic characteristics of the consolidation phase, and success with secondary and tertiary distraction. SUMMARY: After 15 years of clinical use in children for craniofacial deformities, recent advances in distraction osteogenesis have obviated tracheostomy in most newborns with micrognathia and severe airway obstruction. Applications of this technique to children with airway issues related to micrognathia or retrognathia have been rapidly expanding. In addition, refinements in distraction technique have advanced treatment of nonairway-related mandibular deformities. PMID- 15548911 TI - Contemporary diagnostic and management techniques for extraesophageal reflux disease. AB - PURPOSE OF REVIEW: To review recent advances in the diagnosis and treatment of extraesophageal reflux. RECENT FINDINGS: For most patients, the diagnosis of extraesophageal reflux relies on history and laryngoscopic exam. The reliability and reproducibility of reporting these measures may be improved with validated symptom questionnaires and standardized scoring of physical exam findings. Though dual probe pH monitoring has been the gold standard for diagnosis, it does not measure non-acid reflux events. Intraluminal impedance monitoring has the capability of measuring all esophageal reflux events (liquid, solid, gas) and may be useful in the diagnosis of non-acid reflux. Proton pump inhibitors have replaced histamine receptor antagonists as the mainstay of treatment for extraesophageal reflux. Histamine receptor antagonists are used predominantly for nocturnal acid breakthrough, or step-down therapy. Promotility and cytoprotective agents are used less commonly. Baclofen is currently being evaluated for its ability to decrease the incidence of transient lower esophageal sphincter relaxations and reduce post-prandial acid and non-acid reflux events. For individuals refractory to medical therapy, laparoscopic fundoplication techniques have proven efficacy in relieving some symptoms; the long-term benefit is not yet known. There is yet no established data on the effects of endoluminal therapies on extraesophageal reflux symptoms. SUMMARY: The diagnosis of extraesophageal reflux for most patients relies on history and laryngoscopic exam. The diagnosis can be further verified by dual probe pH and impedance monitoring. Proton pump inhibitors are the mainstay of treatment. Laparoscopic fundoplication is proven to relieve symptoms, but there is yet no data on the effects of endoluminal therapies on extraesophageal reflux symptoms. PMID- 15548912 TI - Update on the endoscopic management of laryngeal cancer. AB - PURPOSE OF REVIEW: This update addressing the endoscopic management of laryngeal cancer presents new data published after January 2003. This new information is interpreted in the context of contemporary management principles established before 2003. RECENT FINDINGS: Endoscopic techniques in the management of laryngeal cancer continue to undergo modifications designed to improve oncologic efficacy in a manner that maximally preserves function. The modifications reported over the past two years have included the introduction of new diagnostic techniques, new surgical approaches, new endoscopic equipment, and new management philosophies. SUMMARY: Endoscopic resection has become a widely accepted and commonly employed management option in the treatment of laryngeal cancer. Advances in outcomes analysis have improved the capacity to objectively compare endoscopic treatment to other options. The prospects for applying these tools to a prospective randomized study are limited. Debate will continue as to the role for endoscopic management of laryngeal cancer. PMID- 15548913 TI - Current diagnostic and management trends for recurrent respiratory papillomatosis. AB - PURPOSE OF REVIEW: The purpose of this review is to discuss recent literature regarding diagnostic and management trends for recurrent respiratory papillomatosis (RRP) published within the past year. This includes a discussion of new information regarding the epidemiology and pathogenesis of RRP and an update on adjuvant therapy and new surgical techniques. RECENT FINDINGS: Epidemiological studies have confirmed that juvenile-onset RRP is the most common and most aggressive form of the disease. Age at diagnosis is the most important determinant of disease severity, with younger patients requiring significantly more annual surgeries and more likely to have multicentric disease. Distal tracheal or pulmonary RRP is rare, but carries a significant increase in morbidity and mortality. Research into the pathogenesis of RRP has focused on the genetics of HPV infection and host-virus interactions, suggesting a genetic basis for host susceptibility to RRP. At the present time, surgery remains the mainstay of treatment for RRP. However, recurrence after surgery is common and the search for effective adjuvant therapies is ongoing. The antiviral drug cidofovir has demonstrated efficacy against RRP and is considered a promising new adjuvant treatment of this disease. In an attempt to minimize the untoward effects of surgery, the pulsed-dye laser (PDL) has emerged as a safe and efficacious treatment for select patients with RRP. SUMMARY: While a cure for RRP remains elusive, there has been substantial progress in the diagnosis and management of this disease. Significant advances in clinical and basic science research have dramatically improved our understanding of the epidemiology and pathogenesis of the disease and led to the development of promising new adjuvant therapies and surgical techniques. This has translated to an improved quality of life for many patients with RRP. PMID- 15548915 TI - Contemporary management of spasmodic dysphonia. AB - PURPOSE OF REVIEW: To review current concept in spasmodic dysphonia management. RECENT FINDINGS: The standard of care for spasmodic dysphonia in 2004 remains botulinum toxin chemodenervation for symptomatic management. This is supported by a large body of literature attesting to its efficacy in many different hands, particularly for the adductor form of the disorder. New surgical approaches await the crucial test of time, in view of historical difficulties in achieving long term benefit via recurrent nerve section, and currently, surgery is best reserved for the rare patient who does not benefit or cannot tolerate botulinum toxin injections. Despite efforts to refine both surgical and botulinum toxin treatment, symptom relief in abductor spasmodic dysphonia and dystonia with tremor remains suboptimal. SUMMARY: Spasmodic dysphonia is a disorder of the central nervous system rather than the larynx, and as in other forms of dystonia, interventions at the end organ have not offered a definitive cure. The pathophysiology underlying dystonia is becoming better understood as a result of discoveries in genetically based forms of the disorder, and this approach is the most promising avenue to a long-term solution. PMID- 15548914 TI - Injection laryngoplasty for management of unilateral vocal fold paralysis. AB - PURPOSE OF REVIEW: The purpose of this review is to provide an up-to-date review of injection laryngoplasty technique and currently available injectable materials in the management of unilateral vocal fold paralysis (UVP). RECENT FINDINGS: Many new materials are currently available as substances for injection laryngoplasty. These materials have been developed along distinct of lines reasoning that address the inherent shortcomings of the previously available injectable substances, namely, poor tissue biocompatibility and poor persistence within the larynx. Accordingly, the past decade has seen heightened efforts toward developing implants with improved biocompatibility and longevity. The past year has witnessed publications reporting animal studies and, on occasion, human clinical trials involving the intralaryngeal injection of calcium hydroxyl appetite, autologous fascia, particulate silicone and hyaluronic acid derivatives, and others, for managing glottic insufficiency. SUMMARY: In recent years, the application of injection laryngoplasty to unilateral vocal fold paralysis (UVP) has regained popularity. The technique of injection laryngoplasty has several appealing qualities including relative technical ease, low cost, and wide availability in many clinical settings. A growing number of injectable substances have been developed and tested in the clinical setting of glottic insufficiency. When used to manage unilateral vocal fold paralysis, however, injection laryngoplasty has one irrefutable shortcoming: an inability to address posterior glottic insufficiency. Therefore, while injection laryngoplasty technique becomes increasingly popular for vocal fold augmentation in cases vocal fold paresis, atrophy, and scarring, its role in the treatment of UVP should be limited to cases with an appropriate glottal defect. These techniques should be considered as part of a complimentary armamentarium with framework surgery. PMID- 15548917 TI - Bibliography. Current world literature. Pediatric otolaryngology. PMID- 15548916 TI - Diagnostic tools in laryngology. AB - PURPOSE OF REVIEW: Laryngology has become recognized as a subspecialty within the field of otolaryngology-head and neck surgery over the course of the past 30 years. The evolution of laryngology stems primarily from our better understanding of the physiology of vocal production, society's increased reliance on oral communication in industry, and the resultant demand for improvements in diagnostic and therapeutic protocols to enhance vocal performance. The purpose of this review is to describe current advances in diagnostic tools in laryngology that contribute to our understanding of vocal physiology and our ability to improve quality of life as it relates to vocal performance. RECENT FINDINGS: Diagnostic tools in laryngology allow the clinician the ability to assess the vibratory function of the vocal folds, evaluate laryngeal structure and airway patency, assess glottal and pulmonary airflow, assess the neurologic integrity of the vocal folds, analyze the quality of the voice signal, and assess the degree of functional disability as it relates to laryngeal pathology. SUMMARY: Primary advances in diagnostic laryngology include improvements in laryngoscopy techniques, imaging, electromyography, aerodynamic testing, acoustic analysis, and functional assessment, each of which is improving our ability to understand, diagnose, and treat a wider spectrum of voice, airway, and swallowing disorders. PMID- 15548921 TI - Can better disease definition lead to cure? PMID- 15548922 TI - Toward the definition of cerebral arteriopathies of childhood. AB - PURPOSE OF REVIEW: To facilitate and standardize the diagnosis of cerebrovascular conditions in childhood, particularly in the field of arterial ischemic diseases. RECENT FINDINGS: Progress in diagnostic techniques in the past decade have led to newly established etiologies for childhood stroke, most of which represent some form of vascular pathology. These advances must be integrated into a modern nomenclature system with revised definitions of stroke and arterial wall diseases arteriopathies-in childhood. SUMMARY: This nomenclature system is intended to facilitate and enhance clinical research in childhood stroke, particularly multicenter collaborative studies. PMID- 15548923 TI - Toward a definition of childhood dystonia. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize recent progress toward providing a consistent, sensitive, specific, and useful definition of dystonia as it presents in childhood. RECENT FINDINGS: An NIH-funded consensus group published a definition of childhood dystonia in January of 2003. Recent work has attempted to identify quantitative methods for diagnosis and measurement of childhood dystonia. Techniques include biomechanical, kinematic, and surface EMG measurements that show promise for providing specific and sensitive measures of childhood dystonia. SUMMARY: The results of current research efforts will be useful for verifying and modifying definitions of dystonia to provide consistent and measurable terms for including children in research trials and selecting appropriate interventions for clinical treatment. PMID- 15548924 TI - Toward the definition of childhood migraine. AB - PURPOSE OF REVIEW: The intent of this review is to provide an update on the current understanding of the diagnostic criteria, classification, pathophysiology, evaluation, and management of pediatric migraine. RECENT FINDINGS: The International Headache Society has recently revised its diagnostic criteria and classification system in the 2004 International Criteria for Headache Disorders. These new criteria have incorporated many developmentally sensitive changes that will allow broader application in children and adolescents. Appreciation of the new classification system requires understanding of current views of the pathophysiology of migraine. Migraine is now viewed as an inherited disorder with a primary neuronal initiation of a cascade of neurochemical processes culminating in a spreading wave of cortical neuronal depolarization and regional oligemia. The evaluation of a child with headache begins with a thorough medical history and complete physical and neurologic examination. One of the key questions for the bedside clinician is when to perform further diagnostic studies. Extensive review concludes that the role of further ancillary diagnostic studies, specifically EEG and neuroimaging, is limited. The management of pediatric migraine requires a balance of biobehavioral measures coupled with agents for acute treatment and, if needed, daily preventive medicines. The pharmacologic management of pediatric migraine has been subjected to thorough review, and controlled data unfortunately are limited. The most rigorously studied agents for the acute treatment of migraine are ibuprofen, acetaminophen, and sumatriptan nasal spray, all of which have shown safety and efficacy in controlled trials. For preventive or prophylactic treatment in the population of children and adolescents with frequent, disabling migraine, flunarizine (not available in the United States) is the most efficacious agent, but encouraging data are emerging regarding the use of several antiepileptic agents such as topiramate, disodium valproate, levetiracetam, the antihistamine cyproheptadine, and the antidepressant amitriptyline. SUMMARY: Recent literature has reshaped the definition, evaluation, and management of pediatric migraine. PMID- 15548925 TI - Toward the definition of acute disseminated encephalitis of childhood. AB - PURPOSE OF REVIEW: Acute disseminated encephalomyelitis (ADEM) is a demyelinating disorder most common in childhood and adolescence and thought to have an immune pathogenesis. Some children with ADEM develop additional temporally remote episodes of demyelination with localizations that differ from those of the initial episode and multiple sclerosis is diagnosed. Others have only a single episode. Still others have recurrent episodes in temporal proximity to or with the same localizations as the initial presentation. This review is aimed at delineating what will be required to arrive at a definition of ADEM so as to distinguish it from childhood multiple sclerosis on first presentation. RECENT FINDINGS: All studies to date aimed at differential characterization of ADEM and childhood multiple sclerosis have been retrospective case reviews. Each has examined a different array of historical, physical examination, laboratory, and radiologic characteristics of a differently defined patient population. SUMMARY: Defining ADEM and distinguishing it from multiple sclerosis early in the course of the latter disease depends critically on standardization of prospective, multicenter study design, execution, and data analysis. This has become of paramount importance, as preventive therapies have evolved for recurrent attacks of multiple sclerosis. PMID- 15548927 TI - Anti-IgE therapy in allergic disease. AB - PURPOSE OF REVIEW: Recombinant monoclonal humanized anti-IgE has put forward a fundamentally new concept for the control of allergic disorders. This review will present recent data from clinical studies with anti-IgE in asthma, allergic rhinitis, and food allergy and will examine the place of anti-IgE among current therapeutic options for the treatment of asthma. RECENT FINDINGS: Therapy with anti-IgE depresses circulating free IgE to the limits of detection, inhibits early- and late-phase responses to allergens, suppresses inflammation and improves the control of allergic diseases. In moderate to severe asthma it results in fewer exacerbations and a lower requirement for both corticosteroids and beta-agonists. IgE appears to be an important regulator of high-affinity Fc receptors (FcepsilonRI) and, in the mouse, to enhance mast cell survival and activation. IgE receptors have been found on diverse inflammatory cells. Anti-IgE reduces the expression of FcepsilonRI on inflammatory cells. Current work has documented a marked decrease in tissue eosinophils, lymphocytes, and interleukin 4-positive cells by anti-IgE treatment and has provided insight into the mechanisms underlying improved control of asthma. SUMMARY: Clinical studies with anti-IgE have promoted and will continue to advance the understanding of IgE mediated disease mechanisms. They have documented its efficacy in the treatment of allergic diseases, but much remains to be learned about the most effective clinical strategies and the selection of patients for therapy. PMID- 15548928 TI - Inherited disorders of cytokines. AB - PURPOSE OF REVIEW: Cytokines are soluble mediators involved in the development or function of the immune system. This paper reviews the literature on childhood onset inherited disorders associated with impaired cytokine-mediated immunity. RECENT FINDINGS: Cytokine-mediated immunity defects can be classified into seven different groups: defects in the interleukin (IL)-7 receptor (IL7RA), in the common cytokine receptor gamma chain (gammac) of the IL-2, -4, -7, -15, and -21, and in Jak3 (JAK3) downstream of the gamma chain; mutation in the IL-2 receptor alpha (IL-2RA) and defective expression of the IL-2Rbeta chain; mutations in the gene encoding for a chemokine receptor, CXCR4; mutations in five genes involved in the IL-12/23-interferon-gamma axis (IL12B, IL12RB1, IFNGR1, IFNGR2, STAT1); mutations in three genes involved in the nuclear factor-kappaB signaling pathway (IRAK4, NEMO, IkappaBA); mutations in the tumor necrosis factor receptor signaling pathway (TNFRSF1A); and mutations in the transforming growth factor-1 gene (TGFB1). SUMMARY: Genetic cytokine-mediated immunity defects are associated with a highly heterogeneous group of clinical features, ranging from susceptibility to infections to developmental defects. This heterogeneity highlights the diversity and pleiotropy of cytokines. It is likely that many more cytokine defects and their responsive pathways will be discovered in the coming years, expanding further the heterogeneity associated with this group of childhood-onset illnesses. PMID- 15548929 TI - Understanding asthma pathogenesis: linking innate and adaptive immunity. AB - PURPOSE OF REVIEW: Treatment and even prevention of allergic asthma will require a detailed understanding of disease pathogenesis and in particular identification of factors that govern T-helper type 2 (Th2) immunity. This review defines the priming and differentiation steps necessary to develop antiallergen Th2 immunity and highlights recently identified stimuli that satisfy these requirements. RECENT FINDINGS: Striking discoveries in innate immunity have advanced our understanding of how adaptive immune responses are initiated, yet only recently have these principles been applied to allergic disease. Signaling through certain innate immune receptors, the toll-like receptors (TLR) have been shown to modulate Th2-mediated disease in animal models. The dendritic cell has emerged as the central player in the intricate interplay between the adaptive and innate systems of immunity. Recent studies have also uncovered alternative pathways of initiating allergen sensitization that depend entirely on adaptive, rather than innate immune, triggers. SUMMARY: The adaptive immune system cannot initiate a response without the "permission" of the innate immune system, and this holds true for Th2 responses to aeroallergens, although induction of Th2 immunity in response to TLR signaling varies with the type and dose of TLR ligand. However, under conditions of ongoing Th2 inflammation, the adaptive immune system can act as its own adjuvant and provide the necessary activating signals to initiate an immune response to foreign protein antigens. This may be the mechanism underlying the clinically observed phenomenon of polysensitization in atopic patients and provides another therapeutic target in asthma. PMID- 15548930 TI - Genetic testing will bring interesting times to clinical practice. PMID- 15548931 TI - Rett syndrome: clinical and molecular update. AB - PURPOSE OF REVIEW: New information on the clinical and molecular aspects of Rett syndrome has emerged at an accelerated pace since the identification of mutations in methyl-CpG-binding protein 2 gene (MECP2) was first reported in 1999. Recent reports not only present new insights into the clinical and molecular understanding of this unique disorder but also have important implications for the neurobiology beyond Rett syndrome. RECENT FINDINGS: This review covers recent advances in Rett syndrome from clinical management issues to laboratory-based genetic discoveries. SUMMARY: Clinical management issues include electrocardiographic findings, scoliosis, osteopenia, and motor control. Interesting phenotype-genotype correlations have resulted from the recognition of large-scale deletions in MECP2 as a cause of Rett syndrome, and the more recent identification of an alternate splicing isoform of the gene product. These are beginning to provide a more coherent picture of the spectrum of disease caused by mutations in MECP2. The neurobiologic role of the MECP2 gene in Rett syndrome and normal development has been greatly elucidated with the development of animal models of Rett syndrome and the study of MECP2 in humans and rodents. PMID- 15548932 TI - Genodermatoses 2003-2004. AB - PURPOSE OF REVIEW: Many genodermatoses have been linked in recent years to their respective genes. The underlying biology and integrative nature of these genes with other genes and organ systems is beginning to be understood. This paper reviews recent advances in neurocutaneous disorders, ectodermal dysplasias, and the phenomenon of revertant gene mosaicism. RECENT FINDINGS: In neurofibromatosis type 1, molecular assays are being developed to distinguish malignant from benign and premalignant lesions. Clinical mutation analysis for the NF1 gene has been problematic; a sensitive new assay using automated comparative sequence analysis may be helpful. Revision of clinical care guidelines is ongoing. New data for the prospective management of optic pathway gliomas is reviewed. The two genes that underlie tuberous sclerosis complex, tuberin and hamartin, lie at the center of an important signal transduction pathway with significant implications for pharmacologic treatment. Issues in genetic counseling for this highly variable disease are updated. Extensive progress has been made in understanding the basis of several forms of ectodermal dysplasia. Disorders caused by mutations in p63 and the connexin and NF-kappaB gene families will be reviewed. Finally, phenotypic in vivo amelioration of genodermatoses via revertant gene mosaicism will be discussed as a possible mechanism to be exploited in directed therapeutic approaches. SUMMARY: This paper reviews recent developments in the molecular and biologic bases of neurofibromatosis type 1, tuberous sclerosis, and ectodermal disorders related to p63 and the connexin and NF-kappaB gene families. The concept of revertant gene mosaicism is also discussed as a potential model for gene therapy. PMID- 15548933 TI - Redefining primary pediatric care for children with special health care needs: the primary care medical home. AB - PURPOSE OF REVIEW: As considerations of the quality of health care have matured, the role of pediatric primary care providers and models for the delivery of primary care have received growing attention. Particularly for children with chronic conditions, the need for proactive, planned, and coordinated care delivered in partnership with consumers has become more apparent. The primary care medical home has emerged as a model favored by national organizations representing pediatricians and family physicians as well as national public health policy makers, yet implementation of this model remains limited and the evidence base for its value is not yet highly developed. RECENT FINDINGS: Most studies of primary care outcomes involve individual elements of the medical home such as care coordination and continuity of care. Limited data that are emerging from studies of the medical home model as a whole in practice settings suggest improvements in patient satisfaction and in some areas of utilization. No data are available that examine specific functional or physical health outcomes associated with primary care models like the medical home. SUMMARY: The pediatric primary care medical home provides a care model for both well children and those with special health care needs that expands primary care services beyond those provided in the examination room by individual providers to include systemic services such as patient registries, explicit care planning and care coordination, planned co-management with specialists, patient advocacy, and patient education. There is an immediate need for large-scale, practice-based studies of the outcomes for children and youth, providers, and the health care system when such improvements in primary care are implemented. PMID- 15548934 TI - Current evidence on the impact of continuity of care. AB - PURPOSE OF REVIEW: A core attribute of primary care, continuity of care, is usually viewed as the relationship between a patient and a single practitioner that extends beyond episodes of illness or care for a specific disease and is ongoing over time. The latest evidence of the effectiveness of continuity of care on outcomes for children and adolescents will be presented and placed into the context of prior literature. RECENT FINDINGS: Despite numerous changes in health care delivery systems, recent studies found that clinician continuity continues to be highly valued by patients and their families. From January 1, 2002 to July 1, 2004, five new studies on physician continuity also assessed pediatric outcomes. These studies found an association between physician continuity and lower emergency department use and costs, better coordination of care, and greater patient and parent satisfaction with care. Two new studies on continuity with a primary care practice found that it was associated with higher receipt of recommended developmental, dental, and nutritional advice, as well as up-to-date immunization coverage. SUMMARY: Organizational approaches to maximizing access while maintaining continuity require further development. Efforts to restructure provider reimbursement and information systems need to move from single encounter based systems toward an emphasis on the provision of care over time, to reward and facilitate continuity of care and the positive patient outcomes with which it is associated. PMID- 15548935 TI - Coordination of care for children with special health care needs. AB - PURPOSE OF REVIEW: Coordination of care is an essential function of pediatric primary care, needed most by children with special health care needs (CSHCN). Although complex, its necessity has become better recognized with the recent increase in attention in the United States to the comprehensive "medical home" model of care. RECENT FINDINGS: Coordination is highly dependent on effective communication within the health care system and between the health care system and the larger community. While coordination may best be undertaken at the level of the physician practice, a team approach involving nonphysician staff and families as primary participants may be the best option in many cases. More attention is being paid at the health policy level to the implementation of coordination of care, although solutions to reimbursement barriers have yet to be implemented. Considerable progress on methods to improve care coordination in the primary care practice setting has been made recently. Many of these efforts have used quality improvement techniques adapted from the business world. Emerging measures of the process of care coordination are also being developed, although few studies have been published to date showing a positive impact of care coordination. SUMMARY: The value of coordination of care as an essential part of medical care for children with special health care needs is becoming widely recognized. Methods to implement it within pediatric primary care practices are being developed, although more data demonstrating its value are needed to inform policy changes. PMID- 15548937 TI - Bibliography. Current world literature. Neurology. PMID- 15548936 TI - Medical home, obesity, acute otitis media, and otitis media with effusion. AB - Four areas of pediatric office practice are reviewed: the medical home concept, obesity, acute otitis media, and otitis media with effusion. The concept of the medical home in the care of children with special health care needs, its effect on health care outcomes, and its application to office practice are discussed. The epidemiology and causes of obesity are covered along with options for obesity screening and prevention. Diagnosis and therapy of acute otitis media and otitis media with effusion are reviewed along with discussion of recent practice guidelines for both entities. PMID- 15548938 TI - Benefits and risks of oral contraceptives on cancer. PMID- 15548939 TI - Dietary habits and lung cancer risk among non-smoking women. AB - A case-control study was conducted to investigate the relationship between diet and the risk of lung cancer among women non-smokers and to compare with women smokers in the same population. Data collected by personal interviews from 435 microscopically confirmed cases and 1710 controls were analysed using unconditional logistic regression. In addition to results for all study subjects, associations between diet and lung cancer risk were compared between two highly contrasting groups: smokers (odds ratio (OR) 7.03) and non-smokers (OR 1.00). A protective effect of frequent (daily or several times per week) black tea drinking appeared among non-smoking women (OR 0.65, 95% confidence interval (CI) 0.43-0.99). Among smoking women, protective effects were observed for frequent intake of milk/dairy products (OR 0.56, 95% CI 0.32-0.96), coffee (OR 0.47, 95% CI 0.25-0.88), and wine consumption (daily or weekly OR 0.60, 95% CI 0.37-0.98; monthly OR 0.60, 95% CI 0.39-0.94). Inverse associations with the risk appeared for physical exercise for smokers only, and for the body mass index both among non-smoking and smoking women. Some items of diet may contribute to variation in risk among women in the Czech Republic; their importance seems to vary in relation to their status in smoking, the dominant factor in the aetiology of lung cancer. PMID- 15548940 TI - Phytoestrogen supplementation: a case report of male breast cancer. AB - Patients seeking alternatives to hormone replacement are increasingly using non prescription phytoestrogen supplements. The potential of these herbal remedies to prevent bone loss, heart disease, menopausal symptoms or breast cancer has been a focus of attention in scientific and lay literature. It is important to understand the effects of phytoestrogens, particularly whether excess exposure can promote hyperplasia or neoplasia of breast tissue. We report the case of a man diagnosed with breast cancer whose history was notable for extensive use of supplemental phytoestrogens and the absence of family history of breast cancer or BRCA1/BRCA2 mutation. In conclusion, breast tissue effects of phytoestrogens remain unclear. The increasing popularity and availability of phytoestrogen dietary supplements necessitates additional research in order to counsel patients regarding their safety and efficacy. PMID- 15548941 TI - Breast cancer incidence, mortality and survival in the Umbria region of Italy (1978-1999). AB - Recent incidence, mortality and survival rates of female invasive breast cancer in the Umbria region were compared with data of 20 years ago and with data from other Italian Registries. Incidence data, derived from an ad hoc survey and from cancer registry records, were analysed for the periods 1978-1982 and 1994-1999; mortality trend rates were calculated based on data from official publications and the variation was assessed by joinpoint analysis. Fifteen-year survival rates, observed, relative and age-adjusted, were also calculated. Compared with the years 1978-1982, the age-adjusted incidence rates rose during 1994-1999, while age-adjusted mortality rates decreased in this latter period. Incidence and mortality rates specific for each age group showed a constant increase with age, in both periods of observation, especially among women from 50 to 75 years of age. The 5-year relative survival rates increased 17%, rising from 0.71 in 1978 1982 to 0.86 in 1994-1998; the survival improvement was evident among women aged 45-54, and 65-74. Age-adjusted incidence and mortality rates in Umbria were lower than in other Italian Registries and survival rates showed a better prognosis for breast cancers diagnosed in Umbria in the period 1994-1998. In conclusion, the 1994-1998 decline in breast cancer mortality and the survival improvement may in part reflect the effectiveness of earlier detection by mammography and the progress in treatments. Genetic and lifestyle factors along with the improvement in diagnosis, and in quality of data collection and classification could have determined the recorded increase in breast cancer incidence. PMID- 15548942 TI - Multicenter clinical experience with large core soft tissue biopsy without vacuum assistance. AB - The increasing interest in accurate pretreatment diagnosis of solid tumours by morphology, immunohistochemistry, genetics and molecular biology requires clinicians to obtain undamaged large core biopsies. Simultaneously, medical imaging and surgery give priority to minimal tissue injury, affordable technology and optimal patient compliance. A new large core soft tissue biopsy device has been developed to meet the above criteria. After intensive preclinical testing, 30 patients gave informed consent and 26 underwent the new diagnostic biopsy procedure. The sample was studied by morphology, immunohistochemistry and, where indicated, by molecular biology. Successful diagnosis was considered when in line with clinical follow-up and, as for all malignant lesions, when confirmed by open biopsy or surgery. No difficulties in the technique were encountered in 25 patients. In one patient the procedure was prematurely stopped because of anxiety. In all other 25 procedures a complete diagnosis was reached with regard to morphology, immunohistochemistry and molecular biology. A number of radiologists suggested some automation of the technique. This new large core soft tissue biopsy system performs well in the clinical context without injury to the breast parenchyma or artefacts in the harvested tissue specimen. The system meets almost all of the proposed technical and financial requirements. Automation is underway. PMID- 15548943 TI - Prolonged lactation reduces ovarian cancer risk in Chinese women. AB - To investigate the effect of lactation on the risk of ovarian cancer for Chinese women, a case-control study was conducted in Hangzhou, capital of Zhejiang province, China. Cases were 275 patients with histologically confirmed epithelial ovarian cancer. Controls were 623 women without neoplasm. All participants were parous women who had given at least one live birth and had been residents of Zhejiang province for at least 10 years. Information was collected using a validated and reliable questionnaire on total months of lactation, number of children breastfed, and average duration of lactation per child. Multivariate logistic regression models were used to assess the association between ovarian cancer risk and lactation variables, accounting for age, locality, full-term pregnancy, oral contraceptive use and family history of the cancer. The adjusted odds ratios were 0.51 (95% confidence interval (CI) 0.3-0.9) and 0.44 (95% CI 0.2 0.9) respectively for women with over 12 months of lactation and at least three children breastfed, compared with those with 4 months or less lactation and one child breastfed. The corresponding dose-response relationships were also significant (P<0.05). Therefore, prolonged lactation could reduce the risk of ovarian cancer for Chinese women. PMID- 15548944 TI - Risk of melanoma and vitamin A, coffee and alcohol: a case-control study from Italy. AB - Limited information is available on the association between the risk of cutaneous malignant melanoma (CMM) and dietary factors. The issue was investigated using data from a case-control study conducted in Italy between 1992 and 1994, including 542 patients with incident, histologically confirmed CMM and 538 controls, admitted to the same hospitals as cases for non-dermatologic and non neoplastic diseases. We found a significant inverse association between vitamin A intake and CMM risk. The multivariate odds ratio, after allowance for phenotypic characteristics and history of sunburns, for the highest, compared with the lowest, quartile of intake was 0.71 (95% confidence interval (CI) 0.50-1.02) for beta-carotene, 0.57 (95% CI 0.39-0.83) for retinol, and 0.51 (95% CI 0.35-0.74) for total vitamin A. We found no appreciable association of CMM risk with selected food items, including fish, meat, vegetables, fruit, dairy products, wholemeal bread, alcohol, coffee and tea drinking. Consumption of tea appeared to have a protective effect on CMM risk. The relations with measures of dietary vitamin A are, however, moderate compared with the associations between phenotypic characteristics, sun exposure and number of naevi and CMM risk. PMID- 15548945 TI - Glutathione S-transferase (GST) M1, T1 and N-acetyltransferase 2 (NAT2) polymorphisms and urothelial cancer risk with tobacco smoking. AB - The objective of this study was to examine the association between the genetic polymorphism of glutathione S-transferase (GST) M1, T1 and N-acetyltransferase 2 (NAT2) genes and urothelial cancer risk in relation to smoking status. In this study, 325 Japanese patients with urothelial transitional cell carcinoma and 325 healthy controls were compared for frequencies of GSTM1, T1 and NAT2 genotypes. The frequencies of GSTM1 null genotype and NAT2 slow genotype were significantly higher in the cases than in the controls (adjusted odds ratio (OR) 1.37, 95% confidence interval (CI) 1.01-1.87, adjusted OR 3.09, 95% CI 1.69-5.63, individually). Furthermore, the risk of GSTM1 null genotype and NAT2 slow genotype was higher among smokers (adjusted OR 1.48, 95% CI 1.01-2.15, adjusted OR 4.28, 95% CI 1.96-9.36, individually). The regression analysis of cancer risk as a function of the amount of smoking showed that the susceptibility of people who had GSTM1 null genotype increased from 45 pack-years, while the susceptibility of people with NAT2 intermediate or slow genotype increased rapidly from 25 pack-years, compared with non-smokers. A multiplicative interaction between NAT2 intermediate or slow genotype and pack-years of smoking was found (P<0.001), but GSTM1 null genotype was not (P=0.06). Our results indicate that the GSTM1 null genotype and NAT2 intermediate or slow genotype are associated with an increased risk of urothelial cancer in relation to smoking amounts. Furthermore, the interaction between NAT2 intermediate or slow genotype and pack-years of smoking has a strong impact on urothelial cancer. PMID- 15548947 TI - Baicalin induces NAD(P)H:quinone reductase through the transactivation of AP-1 and NF-kappaB in Hepa 1c1c7 cells. AB - Baicalin (5,6,7-trihydroxyflavone-7-O-D-glucuronic acid, BA) is a flavone isolated from Scutellariae radix. In our previous report BA was a major active principle of NAD(P)H:quinone reductase (QR) induction mediated by Scutellariae radix extract and the induction was related to the transcriptional activation of the QR gene in Hepa 1c1c7 cells. The primary aim of the present study was to determine the molecular mechanism of QR gene expression by baicalin. The antioxidant or electrophile response element (ARE/EpRE) found at the 5'-flanking region of phase II genes may play an important role in mediating their induction by xenobiotics, including chemopreventive agents. In accordance, to study the molecular mechanisms of QR gene expression by BA, electrophoretic mobility shift assay (EMSA), using nuclear extracts of treated and untreated cells against ARE, activator protein-1 (AP-1) or nuclear factor-kappaB (NF-kappaB) binding sites, showed that BA increased the binding levels of the parameters in a dose-dependent manner. Further, Hepa 1c1c7 cells were transiently transfected with a plasmid containing three copies of the AP-1- or NF-kappaB-binding site linked to a chloramphenicol acetyltransferase (CAT) reporter gene. Using the CAT reporter gene assay, a dose-dependent transactivation of AP-1- or NF-kappaB-mediated CAT expression was observed with the treatment of BA. These results clearly indicate that BA induces the QR gene expression and activity by transactivation of AP-1 and NF-kappaB, and thus BA may be considered as a potential cancer chemopreventive agent with the induction of phase II detoxification enzyme. PMID- 15548946 TI - Suppression of diethylnitrosamine and 2-acetylaminofluorene-induced hepatocarcinogenesis in rats by tocotrienol-rich fraction isolated from rice bran oil. AB - The anticancer efficacy of tocotrienol-rich fraction (TRF) was evaluated during diethylnitrosamine (DEN)/2-acetylaminofluorene (AAF)-induced hepatocarcinogenesis in male Sprague-Dawley rats. TRF treatment was carried out for 6 months, and was started 2 weeks before initiation phase of hepatocarcinogenesis. Morphological examination of the livers from DEN/AAF rats showed numerous off-white patches and few small nodules, which were significantly reduced by TRF treatment. Cytotoxic damage by DEN/AAF was estimated by alkaline phosphatase (ALP) release into the plasma from the cell membranes. DEN/AAF caused a twofold increase in the activity of ALP in plasma as compared with normal control rats, and this increase was prevented significantly by TRF treatment. We observed an increase of 79% in liver ALP activity in DEN/AAF rats, which was further increased by another 48% after the administration of TRF. Hepatic activity of glutathione S-transferase (GST) was also increased (3.5-fold) during the induction of hepatic carcinogenesis. Lipid peroxidation and low-density lipoprotein (LDL) oxidation increased threefold following initiation by DEN/AAF as compared with normal control rats. However, TRF treatment to DEN/AAF-treated rats substantially decreased (62-66%) the above parameters and thus limited the action of DEN/AAF. We conclude that long-term intake of TRF could reduce cancer risk by preventing hepatic lipid peroxidation and protein oxidation damage due to its antioxidant actions. PMID- 15548948 TI - The transposability of the Mediterranean-type diet in non-Mediterranean regions: application to the physician/allied health team. AB - Studies are consistently declaring that the Mediterranean-type diet is transposable to non-Mediterranean regions. The nutritional end points of Med-type eating appear to be achievable through foods from a variety of traditions and appear to support predetermined expectations surrounding food preparation, choice, taste and sensory appeal. The broad emphasis on minimally processed plants and their products (vegetables, fruit, legumes, wholegrains, nuts, seeds and oils); low fat dairy, fish, less emphasis on animal products and removal of partially hydrogenated fats has piqued the attention of health professionals who are interested in arresting the incidence of chronic disease. The theoretical underpinnings of Med-type eating have driven new understandings in dietary guidelines, which is especially timely as well-marketed fad diets loom large on the current health horizon. PMID- 15548949 TI - 1997--A curious year in Sweden. PMID- 15548950 TI - Human bonding. PMID- 15548952 TI - Parallel Imaging: (Part II). PMID- 15548953 TI - SMASH, SENSE, PILS, GRAPPA: how to choose the optimal method. AB - Fast imaging methods and the availability of required hardware for magnetic resonance tomography (MRT) have significantly reduced acquisition times from about an hour down to several minutes or seconds. With this development over the last 20 years, magnetic resonance imaging (MRI) has become one of the most important instruments in clinical diagnosis. In recent years, the greatest progress in further increasing imaging speed has been the development of parallel MRI (pMRI). Within the last 3 years, parallel imaging methods have become commercially available, and therefore are now available for a broad clinical use. The basic feature of pMRI is a scan time reduction, applicable to nearly any available MRI method, while maintaining the contrast behavior without requiring higher gradient system performance. Because of its faster image acquisition, pMRI can in some cases even significantly improve image quality. In the last 10 years of pMRI development, several different pMRI reconstruction methods have been set up which partially differ in their philosophy, in the mode of reconstruction as well in their advantages and drawbacks with regard to a successful image reconstruction. In this review, a brief overview is given on the advantages and disadvantages of present pMRI methods in clinical applications, and examples from different daily clinical applications are shown. PMID- 15548954 TI - Parallel imaging at high field strength: synergies and joint potential. AB - MRI faces fundamental limitations in terms of sensitivity and speed. These limitations can be effectively tackled by the transition to higher field strengths and parallel imaging technology. Owing to largely independent physics, the two approaches can be readily combined. Considering the specific advantages and disadvantages of high field strength and parallel imaging, it is found that the combination is particularly synergistic. In the joint approach, the two concepts play different roles. Higher field strength acts as a source of higher baseline signal-to-noise ratio (SNR), while parallelization acts as a means of converting added SNR into a variety of alternative benefits. This interplay holds promise for a broad range of clinical applications, as recently illustrated by several imaging studies at 3 T. As a consequence, clinical MRI at 3 T and higher is expected to rely more on parallel acquisition than at lower field strength. The specific synergy with parallel imaging may even make 3 T the field strength of choice for a range of exams that conventionally work best at 1.5 T or less. PMID- 15548955 TI - Improved perfusion and tracer kinetic imaging using parallel imaging. AB - Dynamic contrast enhanced MRI is a valuable method to get functional information of the microvessel system. This article will describe the improvement in perfusion and tracer kinetic imaging by using parallel imaging techniques. The basic principles of kinetic analysis, the contrast mechanisms used and the scan techniques applied are addressed to give a better understanding of the influence of parallel imaging on dynamic contrast enhanced MRI. The effect of different parallel imaging techniques on scan efficiency and scan planning is part of the discussion. From the clinical applications shown, it can be concluded that parallel imaging is a very important contribution for improved dynamic contrast enhanced imaging. PMID- 15548956 TI - Parallel imaging techniques in functional MRI. AB - Originally developed for increased scanning velocity in cardiac imaging, parallel imaging (PI) techniques have recently also been applied for the reduction of artifacts in single-shot techniques. In functional brain imaging (fMRI) techniques, PI has been used for several purposes. It has been applied to reduce the distortions caused by the length of the echo-planar imaging readout, diminution of the gradient-related acoustic noise, as a means to increase acquisition speed or to increase the achievable brain coverage per unit time. In this article, the different applications of PI techniques in fMRI are reviewed, together with the basic theoretical background and the recently developed hardware necessary to achieve rapid, high signal-to-noise ratio PI-fMRI. PMID- 15548957 TI - Artifact reduction using parallel imaging methods. AB - Multiple receiver coils produce images with different but complementary views of a patient. This can be used to shorten scans times but there often remain image artifacts caused by patient motion or physiological processes such as flowing blood. This paper reviews how the extra information from the multiple coils can be used to reduce image artifacts. In one method, affected portions of data can be identified and discarded but enough information is still available to reconstruct an improved image. In other methods, the motion itself is determined and the corrupted data is then corrected, leading to an image with reduced artifacts. Results are presented from images corrupted by motion or by flowing blood. PMID- 15548958 TI - Recipient death during a live donor liver transplantation: who gets the "orphan" graft? AB - The limited availability of deceased organ donors, prolongation of waiting time, and increasing number of patients dying awaiting transplantation have contributed to the increased use of adult-to-adult living-donor liver transplant. In the event that the intended recipient dies after the donor graft has been procured but before it has been transplanted, what should be done with the graft? A structured, nine-item oral survey of 26 experts in liver transplantation was conducted in June and July 2003. Respondents were selected primarily because of their extensive experience with liver transplantation, especially adult-to-adult living-donor transplant. All respondents said the surgical team should try to use the available graft for another recipient. Twenty-one respondents believed consent from the donor or the donor's family was required for allocation, whereas 19% believed consent desirable but not required. Nine respondents recommended an allocation organization place the graft, whereas 17 respondents recommended placement within the donor hospital. Two of the respondents had previously encountered this situation, whereas four had experienced an intraoperative recipient death before procurement of a live donor graft. On the basis of the responses, we offer the following recommendations for handling orphan liver grafts: (1) obtain predonation informed consent from all donors that indicates what the donor would want to have done with the "orphan graft" in all cases of living-donor liver transplantation; (2) avoid the premature removal of the donor graft until the recipient hepatectomy and survival are likely; (3) if a live donor graft has been procured and cannot be transplanted into the intended recipient, and if informed consent has been obtained before the donor operation, the organ should be reallocated without delay to minimize cold ischemia time and maximize the utility of the graft. PMID- 15548959 TI - Involvement of interleukin-18 in acute graft-versus-host disease in mice. AB - BACKGROUND: Interleukin (IL)-18 stimulates T helper 1 (Th1)-mediated immune responses and the development of cytotoxic T lymphocytes (CTLs). Antihost CTLs are major effectors in acute graft-versus-host disease (aGvHD), a potentially fatal complication after allogeneic stem-cell transplantation. We investigated the relevant role of IL-18 in the development of aGvHD in mice. METHODS: Irradiated (C57BL/6x DBA/2) F1 (BDF1) mice transplanted with wild-type (WT) C57BL/6 (B6) splenocytes were compared with those transplanted with IL-18Ralpha deficient B6 splenocytes with respect to Th1 development, CTL activity, severity of aGvHD, and survival. RESULTS: Transplantation of WT B6 spleen cells into BDF1 mice induced aGvHD that was accompanied by elevation of both serum IL-18 levels and IL-18 receptor alpha chain (IL-18Ralpha) expression on engrafted T cells. The transplantation of WT B6 cells also induced high antihost CTL activity in host spleen, whereas transplantation of IL-18Ralpha-deficient B6 cells exhibited significantly reduced antihost-specific CTL activity, indicating that IL-18Ralpha deficient CTLs were less cytotoxic than IL-18Ralpha-expressing CTLs. Moreover, the hosts receiving transplants with the IL-18Ralpha-deficient B6 cells had fewer fatal tissue injuries and increased their survival rates as compared with those receiving transplants with WT cells. Nevertheless, Th1 development in the hosts was the same, regardless of the type of donor cells. CONCLUSIONS: These results suggest that Th1 induction and baseline CTL activity in aGvHD occur in the absence of IL-18, but endogenous IL-18 further accelerates aGvHD reaction to its full-blown manifestation. Thus, IL-18 may be involved in the development aGvHD by enhancing CTL activity. PMID- 15548960 TI - Donor brain death aggravates chronic rejection after lung transplantation in rats. AB - BACKGROUND: Many recipients of lung transplants from brain-dead donors develop bronchiolitis obliterans, a manifestation of chronic rejection. It has been shown that brain death increases inflammatory mediators and accelerates acute rejection in kidney, liver, and heart transplants. In this study, the authors investigated the hypothesis that brain death increases inflammatory mediators in the donor lung and subsequently aggravates chronic rejection of the lungs after transplantation in rats. METHODS: Brain death was induced in F344 rats by inflation of a subdurally placed balloon catheter. After 6 hr, donor lungs were assessed for influx of leukocytes, expression of cell adhesion molecules, and cytokine mRNA expression. For assessment of the lung after transplantation, lungs from brain-dead F344 rats were transplanted into WKY rats. Lung function after transplantation was monitored by chest radiographs during an observation period of 100 days. At the end of this period, the lungs were histologically examined; also, cytokine mRNA expression was measured. Lungs from ventilated living donors and living donors served as controls. RESULTS: After 6 hr of brain death, influx of polymorphonuclear cells and macrophages and expression of vascular cell adhesion molecule-1 in the donor lungs was increased. After transplantation at postoperative day 100, the lung function was significantly decreased compared with allografts from living donors. In the lung allografts from brain-dead donors, histologic symptoms of chronic rejection were obvious, including severe intimal hyperplasia but without bronchiolitis obliterans. Interleukin-2 mRNA was significantly increased in allografts from brain-dead donors compared with living donors. CONCLUSIONS: This study shows that brain death induces an inflammatory response in the donor lung and subsequently aggravates chronic rejection after transplantation. This may explain the clinical difference in long-term function between lungs from cadaveric donors and living donors. PMID- 15548961 TI - Induction of Fractalkine and CX3CR1 mediated by host CD8+ T cells in allograft tolerance induced by donor specific blood transfusion. AB - BACKGROUND: Donor-specific tolerance to heart allografts in the rat can be achieved by donor-specific blood transfusions (DST) before transplantation. This tolerance induction requires the presence of host CD8 T cells and is characterized by the infiltration of numerous leukocytes. METHODS: To identify new mediators involved in tolerance induction, gene searching was performed and resulted in the identification of the Fractalkine receptor, CX3CR1, as being highly expressed in tolerated allografts. RESULTS: We showed that the high CX3CR1 mRNA accumulation found in tolerated allografts was related to the active recruitment of monocytes/macrophages. CX3CR1 transcript accumulation was preceded by an early expression of its ligand, Fractalkine, by graft endothelial cells. Interestingly, depletion of recipient CD8 cells led to a dramatic decrease in both CX3CR1 and Fractalkine mRNA levels. Moreover, in vitro, CD8 T cells from DST primed animals were found to strongly induce Fractalkine expression in an allogeneic endothelial cell line. CONCLUSION: This is the first report describing Fractalkine, a chemokine usually described in inflammatory processes, as being expressed in a model of allograft tolerance. PMID- 15548962 TI - Caspase 3 inhibition improves survival and reduces early graft injury after ischemia and reperfusion in rat liver transplantation. AB - BACKGROUND: Apoptosis plays a crucial role after ischemia-reperfusion in organ transplantation. It is executed by caspases and influenced by the rheostat of pro and anti-apoptotic proteins of the bcl-2 family. This study investigated the effect of specific inhibition of caspases 3 and 7 on graft function, survival, and hepatic bcl-2 levels after liver transplantation. METHODS: Lewis rats underwent syngeneic orthotopic liver transplantation after 16 hr of cold graft storage (in University of Wisconsin solution). Livers of donor animals treated with D(OMe)E(OMe)VD(OMe)-fluoromethylketone (specific inhibitor of apoptosis executor caspases 3 and 7), and appropriate control groups, were investigated. Early graft injury was quantified by measurement of bile flow and determination of microvascular graft injury by using in vivo fluorescence microscopy. Apoptosis and its regulation were examined by terminal deoxynucleotide transferase-mediated dUTP nick-end labeling staining and Western blot analysis of cell death effectors, respectively. RESULTS: After specific in vivo caspase inhibition, Western blot analysis revealed inhibition of caspase-induced cleavage of poly-ADP ribose-polymerase. Inhibition of caspases 3 and 7 resulted in a significantly decreased number of apoptotic endothelial cells and improved microvascular perfusion. A cell protective effect was also suggested by an increase of bcl-2 levels at 7 days. Most important, specific caspase blockade resulted in improved rat survival after liver transplantation. CONCLUSION: Specific inhibition of apoptosis executor caspases effectively reduces graft ischemia-reperfusion injury and improves survival in liver transplantation. Better tissue preservation after caspase inhibition correlates with reduced apoptosis execution, improved microvascular perfusion, and bcl-2 up-regulation. Therefore, specific caspase inhibition represents a promising regimen for clinical use in liver transplantation. PMID- 15548963 TI - Chimerism and tolerance post-in utero transplantation with embryonic stem cells. AB - BACKGROUND: Clinical application of in utero transplantation (IUT) in human fetuses with intact immune systems resulted in a very low level of donor chimerism. In this study, we examined whether the fetal immune system early in the second trimester of pregnancy (13.5 dpc) can initiate immune tolerance for major histocompatibility complex (MHC)-mismatched embryonic stem (ES) cells. We also examined whether immune tolerance mechanisms respond differently to ontogenetically different stem cells. METHODS: MHC-mismatched ES, fetal liver (FL), and bone-marrow (BM) cells (H-2kd) at 1 x 10(9) cells/kg fetal body weight were injected intraperitoneally into 13.5 dpc BALB/c fetuses (H-2Kd). Peripheral chimerism was determined in blood by flow cytometry (sensitivity< or =0.1%) at monthly intervals. Donor-specific immune responses were determined by cytotoxic lymphocyte (CTL) assay, mixed lymphocyte reaction, and T helper (Th)1 and Th2 cytokine assays. Chimeric mice at the age of 9 months received postnatal boosts (PB) with minimal conditioning of 200 cGy by intravenous injection of 1 x 10(9) of the corresponding cells/kg body weight. RESULTS: After IUT with ES, FL, or BM cells, the level of peripheral chimerism within the first 9 months of life was 0% to 0.4%. PB with 1 x 10(9)/kg of corresponding cells resulted in a decrease in the peripheral chimerism to 0% within 2 weeks of PB. CTL and cytokine assays before and after PB demonstrated a shift toward immunity. CONCLUSIONS: Immunologic tolerance was not achieved after IUT of murine fetuses at 13.5 dpc with MHC-mismatched ES cells, and only a low level chimerism was achieved. PMID- 15548964 TI - E-selectin early overexpression induced by allogeneic activation in isolated mouse lung. AB - BACKGROUND: The interaction between host lymphocytes and graft endothelial cells plays an important role in graft rejection. METHODS: Using our model of isolated ventilated lung from female mouse perfused with fresh blood from either isogeneic or allogeneic male mouse for 3 hours without noticeable ischemia, we have investigated the kinetics of the early events after endothelial cell triggering by E-selectin engagement. RESULTS: Isogeneic perfusion induced nonspecific endothelial cell activation, which was characterized by up-regulation of E selectin, intercellular adhesion molecule (ICAM)-1, and of the pro-inflammatory cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-2, and lymphotoxin alpha (mRNAs by real-time polymerase chain reaction). Allogeneic perfusion was characterized after 3 hours by an additional loose adhesion of lymphocytes mediated by the E-selectin and related to the allogeneic activation of endothelial cells. These in turn expressed the I-A molecule (immunostaining). ICAM-1 and lymphocyte function-associated antigen (LFA)-3 mRNA levels were significantly increased in lung extracts after 2 hours, then vascular cell adhesion molecule (VCAM)-1 and TNF-alpha mRNAs after 3 hours without evidence of TNF-alpha production (enzyme-linked immunoadsorbent assay). The major participation of the E-selectin in early allogeneic activation by way of the protein kinase (PK)C pathway was confirmed by using a neutralizing anti-CD62E monoclonal antibody or the inhibitory PKC 19-31 fragment. CONCLUSIONS: Altogether, our results demonstrate that E-selectin expression (1) is not a consequence of TNF-alpha triggering, (2) up-regulates its own expression and expression of I-A, VCAM-1, TNF-alpha, and lymphotoxin-alpha mRNAs, and (3) down regulates expression of LFA-3 and ICAM-1 mRNAs. In conclusion, in our physiologic model, the E-selectin highly participates in the loose adhesion of allogeneic lymphocytes and in the early activation of endothelial cell and therefore in structural and functional lung alterations. PMID- 15548965 TI - Targeted T-cell depletion or CD154 blockade generates mixed hemopoietic chimerism and donor-specific tolerance in mice treated with sirolimus and donor bone marrow. AB - BACKGROUND: The administration of donor specific bone marrow (DSBM) to mice conditioned with antilymphocyte serum (ALS) and sirolimus can result in stable multilineage mixed chimerism and long-term graft survival. This study seeks to determine if either the targeted depletion of CD4 and/or CD8 pos T cells or costimulation blockade can substitute for ALS and preserve the efficacy of this regimen. METHODS: C57BL/6 recipients of BALB/c skin allografts were treated with DSBM (150 x 10(6) cells), sirolimus (24 mg/kg intraperitonealy), and either ALS or various monoclonal antibodies (alphaCD4, alphaCD8, alphaCD154 alone or in combination). Recipient peripheral blood mononuclear cell (PBMC) depletion, donor chimerism, and deletion of donor reactive T cells were assessed using flow cytometry. The specificity of immunologic nonreactivity and the presence of immunoregulatory activity were assessed through a mixed lymphocyte reaction assay. RESULTS: The administration of ALS, sirolimus, and DSBM resulted in sustained recipient PBMC depletion, transient chimerism, and prolonged graft survival. The substitution of an equivalent degree and duration of targeted depletion of either CD4 or CD8 pos T cells alone for ALS failed to produce chimerism or prolonged graft survival. In contrast, depletion of both CD4 and CD8 pos T cells resulted in durable multilineage chimerism, indefinite allograft acceptance (>350 days), and donor-specific tolerance to secondary skin grafts. Substitution of alphaCD154 monoclonal antibody for ALS also resulted in a state of mixed chimerism and donor specific tolerance. This tolerant state appears to be maintained at least partially through clonal deletion and suppression. CONCLUSION: Either combined CD4 and CD8 T-cell depletion or alphaCD154 blockade can effectively substitute for ALS in producing chimerism and tolerance in this model. PMID- 15548966 TI - Urgent need for normalization of corneal graft quality controls in French eye banks. AB - BACKGROUND: Assessment of corneal tissue quality before graft is mainly based upon the determination of endothelial cell density (ECD) by eye banks. These cells are responsible for corneal transparency, and ECD correlates with graft survival. In France and often elsewhere in Europe, ECD is measured using a "naked eye" procedure under a light microscope. To measure objectively the reliability of ECD determination in France, we developed four test corneas with a known ECD. METHODS: The test corneas consisted of 1 mm2 of human corneal endothelium with stained cell borders. The 64 technicians of the 21 French eye banks counted according to the protocol applied in their respective centers. RESULTS: More than half of the 256 counts (152, 59%) deviated by more than 10% from actual ECD. Of the counts, 85 (33%) were over-estimated, and 67 (26%) were under-estimated. Deviation ranged between 42% under-estimation and 82% over-estimation. Eight banks (38%) constantly over-estimated, and nine (43%) under-estimated ECD. Half of the inter-technician gaps within an eye bank were more than 10%, with a maximum of 51%. CONCLUSIONS: This audit highlights the unacceptable lack of reliability of manual ECD assessment in French eye banks. This surely indicates the delivery of poor quality corneas for graft in certain centers and wastage in others. We urgently advocate normalization of French counting methods. This may require upgrading to a computer-aided method. PMID- 15548967 TI - Treatment of established recurrent hepatitis C in liver-transplant recipients with pegylated interferon-alfa-2b and ribavirin therapy. AB - INTRODUCTION: The management issues of transplant patients with hepatitis C virus (HCV) are complex, and interferon therapy is often ineffective. We present data from a retrospective review in liver-transplant recipients suffering from HCV recurrence that were treated with pegylated alpha-2b interferon and ribavirin. METHODS: A retrospective review of transplant recipients that received combination pegylated alpha-2b interferon (1.5 mcg/kg/wk) and ribavirin (400-600 mg/day) therapy intended for at least 48 weeks. Complications were recorded and included neutropenia (<750 cells), anemia (hemoglobin <8 g) with and without treatment consisting of blood transfusions, erythropoietin, or dose reduction of ribavirin, and depression. The diagnosis of HCV recurrence was determined by an increase in liver chemistries, histopathologic findings with inflammation along with viral recurrence using the COBAS AMPLICOR HCV test. RESULTS: Fifty-seven liver-transplant recipients were included, 29 naive (group 1) to therapy and 28 nonresponders (group 2) to at least 6 months of interferon and ribavirin therapy. Eight (27.6%) patients in group 1 and six (21%) patients in group 2 were HCV nondetectable at the end of 48 weeks of therapy. Ribavirin therapy was decreased in 13 of 29 (45%) for group 1 and 11 of 28 (39%) in group 2. Therapeutic interventions were 4 of 57 (7%) blood transfusions, 23 of 57 (40%) erythropoietin, and 17 of 57 (30%) filgrastim. CONCLUSION: Combination pegylated interferon with ribavirin appears to effective therapy in HCV recurrence and in HCV nonresponsive to interferon and ribavirin. This data reveals the difficulty and caution that must be taken when treating HCV-R liver-transplant recipients with combination pegylated alpha-2b interferon and ribavirin therapy. PMID- 15548968 TI - Preemptive therapy for hepatitis C virus after living-donor liver transplantation. AB - BACKGROUND: Living-donor liver transplantation (LDLT) is important for patients with end-stage viral hepatitis because of the cadaveric organ shortage. Preliminary results, however, indicate that LDLT might be disadvantageous for patients positive for hepatitis C virus (HCV). METHODS: The subjects were 23 patients who underwent LDLT for HCV cirrhosis. All the patients preemptively received antiviral therapy consisting of interferon-alfa2b and ribavirin, which was started approximately 1 month after the operation. The therapy continued for 12 months after the first negative HCV RNA test. The patients were then observed without the therapy for 6 months (group 1). The therapy was continued for at least 12 months even when the HCV RNA test remained positive (group 2). The subjects were removed from the protocol if they could not continue the therapy for 12 months because of adverse effects or could not start the therapy because of early death. RESULTS: Eight patients were removed from the protocol. Nine patients were assigned to group 1 and the other six to group 2. The sustained virologic response ratio was 39% (9 of 23). There was a significant difference between the groups in the histologic activity score 1 year after the therapy. The cumulated 3-year survival of the HCV-positive patients was 85%, which was comparable with that of patients negative for HCV (n=93 [90%]). CONCLUSIONS: The present preemptive antiviral protocol after LDLT is safe and might warrant a controlled study for confirming its benefit on graft survival. PMID- 15548969 TI - Early cyclosporine a withdrawal in kidney-transplant recipients receiving sirolimus prevents progression of chronic pathologic allograft lesions. AB - BACKGROUND: Nephrotoxicity of calcineurin inhibitors (CNIs) is partially responsible for the development of chronic allograft nephropathy (CAN). Sirolimus has demonstrated its potential to substitute for CNIs because it lacks significant nephrotoxicity and shows a short-term immunosuppressive capacity comparable with that of cyclosporine. This results in the maintenance of better renal function when cyclosporine is eliminated, but it has not been demonstrated whether this benefit is associated with an improvement in the pathologic substrate and a reduction in CAN. METHODS: We analyzed pretransplant and 1-year renal-allograft biopsies from 64 patients enrolled in a multicenter trial. Patients received cyclosporine and sirolimus during the first 3 months after transplant and were then randomly assigned to continue with cyclosporine or have it withdrawn. Histologic chronic allograft lesions were compared between groups. RESULTS: The percentage of patients in whom chronic pathologic lesions progressed was lower in the group of cyclosporine elimination. Significant differences were observed in chronic interstitial and tubular lesions (70% vs. 40.9% [P<0.05] and 70% vs. 47.8% [P<0.05], respectively), whereas no differences were observed in acute lesions (subclinical rejection). Prevalence of CAN at 1 year was lower in this group, as was the severity and incidence of new cases (P<0.05). CONCLUSIONS: Early cyclosporine withdrawal associated with sirolimus administration is followed by an improvement in renal function, a reduction in the progression of chronic pathologic allograft lesions, and a lower incidence of new cases and severity of CAN during the first year after transplantation. This benefit may result in better long-term graft outcome. PMID- 15548970 TI - Urolithiasis after kidney transplantation in pediatric recipients: a single center report. AB - BACKGROUND: Urolithiasis occurs in approximately 6% of adult kidney transplant (KTx) recipients. Limited data are available on urolithiasis after pediatric KTx. We report the incidence, management of, and risk factors for stone development in children after KTx. METHODS: We reviewed the medical records of 399 children who received KTx at our center between September 1986 and January 2003. Transplant outcomes were compared in stone formers and controls. RESULTS: Twenty (5%) patients, age 9+/-5 (X +/- SD) years, developed stones over the follow-up period (74+/-53 months). Time to stone presentation was 19+/-22 months post-KTx. Presenting features were urinary tract infection (UTI), 8; gross hematuria, 5; microscopic hematuria, 2; dysuria without infection, 6; difficulty voiding, 3; and silent stones, 2. Stones were removed by cystoscopy in 11 (55%) patients. Stone composition was determined in 11 patients: calcium phosphate (55%), calcium oxalate (18%), mixed calcium phosphate and oxalate (9%), and struvite (18%). Factors predisposing to stones in study patients included suture retention (n = 4), elevated urinary calcium excretion (n = 2), recurrent UTI (n = 2), and urinary stasis (n = 2). The incidence of UTI was higher (P = 0.003) and of acute rejection was lower (P = 0.02) in stone patients compared with controls. Patient and graft survival rates and the incidence of chronic rejection did not significantly differ between study patients and controls (P = NS). CONCLUSIONS: Urolithiasis is not uncommon in pediatric KTx patients. Factors associated with post-KTx urolithiasis include retention of suture material, recurrent UTI, hypercalciuria, and urinary stasis. Treatment is associated with excellent outcome and low recurrence rate. PMID- 15548971 TI - Evidence for a role of toll-like receptor 4 in development of chronic allograft rejection after cardiac transplantation. AB - BACKGROUND: Long-term success of cardiac transplantation is limited by various forms of graft rejection. The specific mechanisms initiating and controlling these highly specialized immune processes remain unclear so far. METHODS: To investigate the role of innate immunity in the development of allograft rejection, we assessed toll-like receptor (TLR)4 expression and typical downstream effects of TLR signaling (B7-1, interleukin [IL]-12, tumor necrosis factor [TNF]-alpha) in circulating CD14+ monocytes in 38 transplant recipients 1 to 3 years and in 10 transplant recipients 6 to 10 years after transplantation and compared them with 20 healthy controls using reverse transcription polymerase chain reaction, flow cytometry, and enzyme-linked immunoadsorbent assay. The results were matched with endothelial function testing as an early clinical indicator of transplant vasculopathy early after transplantation. RESULTS: Allograft endothelial dysfunction (ED) was defined as a compromised coronary flow reserve (CFVR) to acetylcholine (CFVR<2 in 13 of 38 transplant recipients). In these patients, mRNA transcript levels for TLR4 (P<0.05) and surface expression of TLR4 (P<0.005) and B7-1 (P<0.05) on circulating monocytes as well as secretion of IL-12 (P<0.02) and TNF-alpha (P<0.05) were significantly higher than in the remaining 25 patients without ED. Compared with the controls, recipients late after transplantation did not show significantly elevated levels of TLR4 or dependent mediators. These results were compared with mRNA levels in a mice model of acute and chronic rejection. Rejecting mice exhibited elevated mRNA levels for mTLR4 and mB7-1. CONCLUSIONS: Our results suggest activation of innate immunity in heart-transplant recipients through TLR4 contributes to the development of chronic rejection after cardiac transplantation. PMID- 15548972 TI - Everolimus and reduced-exposure cyclosporine in de novo renal-transplant recipients: a three-year phase II, randomized, multicenter, open-label study. AB - BACKGROUND: Everolimus and cyclosporine (CsA) exhibit synergistic immunosuppressive activity when used in combination. We explored the use of everolimus with a CsA-sparing strategy in de novo renal-transplant recipients. METHODS: A phase II, randomized, open-label 3-year study was performed in 111 patients to compare the efficacy and tolerability of everolimus (3 mg/day) in combination with basiliximab, steroids, and either full-dose Neoral (FDN) or reduced-dose Neoral (RDN) (CsA trough levels 125-250 ng/mL and 50-100 ng/mL, respectively). Efficacy failure (biopsy-proven acute rejection, death, graft loss, or loss to follow-up), safety, and renal function were evaluated at 6, 12, and 36 months. A protocol amendment allowed further reduction of CsA exposure after 12 months. RESULTS: Efficacy failure was significantly higher in FDN than in the RDN group at 6 (15.1% vs. 3.4%; P=0.046), 12 (28.3% vs. 8.6%; P=0.012), and 36 (35.8% vs. 17.2%; P=0.032) months. Mean creatinine clearance was higher in the RDN group at 6 (59.7 mL/min vs. 51.1 mL/min; P=0.009), 12 (60.9 mL/min vs. 53.5 mL/min; P=0.007), and 36 (56.6 mL/min vs. 51.7 mL/min; P=0.436) months. Discontinuations and serious adverse events were more frequent in the FDN group. Reduction of CsA exposure for 6 months during the amendment improved renal function in the FDN group. CONCLUSIONS: In de novo renal-transplant recipients, the regimen of everolimus plus RDN was well tolerated, with low efficacy failure and better renal function in comparison with everolimus plus FDN. PMID- 15548973 TI - CXCR3 and CCR5 positive T-cell recruitment in acute human renal allograft rejection. AB - BACKGROUND: Experimental studies suggest that the infiltration of activated T cells into the allograft, the key event in the development of acute renal allograft rejection, depends on the expression of chemokines and their interaction with chemokine receptors expressed on T cells. METHODS: For a more detailed comprehension of the pathogenesis of T-cell recruitment in human acute rejection, the in situ expression of chemokines and chemokine receptors in allografts of 26 patients between day 3 and 9 after renal transplantation was examined in the present prospective study. RESULTS: Immunohistochemical staining showed a significantly increased number of CXCR3 (P<0.01) and CCR5 positive T cells (P<0.01) in the tubulointerstitium of patients with acute allograft rejection according to Banff grade Ia-IIb. Likewise the intrarenal RNA expression of the CXCR3 ligands IP-10 (5.2-fold) and I-TAC (7.2-fold) and the CCR5 ligand RANTES (5.7-fold), was significantly up-regulated in rejecting organs. In situ hybridization revealed that IP-10 but not I-TAC was predominantly expressed by infiltrating leukocytes in the tubulointerstitial area, co-localizing with CXCR3 positive T cells. To a lesser degree expression by tubular cells could be detected, providing a possible explanation for the increased urinary IP-10 excretion we found in patients with rejecting organs. CONCLUSIONS: These data from a prospective, biopsy-controlled study indicate that the local expression of IP-10 and RANTES leads to the directional movement of activated CXCR3 and CCR5 bearing T cells into the renal allograft and mediates acute rejection. Our data provide a rationale that blocking CXCR3 and CCR5 may offer a unique therapeutic approach to prevent episodes of acute rejection in the early phase after renal transplantation. PMID- 15548974 TI - The burden of acute renal failure in nonrenal solid organ transplantation. AB - BACKGROUND: Recipients of nonrenal solid organ transplants are at risk for acute renal failure resulting from cardiac or hepatic failure, prolonged surgery, and nephrotoxic effects of immunosuppression. Single-center studies have suggested a variable incidence of acute renal failure in this population, with an associated increase in mortality. This study examines the incidence of acute renal failure and its associated mortality and morbidity in a modern multicenter cohort. METHODS: All adult liver, heart, and lung transplant recipients from 2002 were identified from the New York Statewide Planning and Research Cooperative System database. The impact of acute renal failure on mortality, length of stay, and charges was analyzed using multivariate regression models. RESULTS: Among 519 liver, heart, and lung transplant recipients, the incidence of acute renal failure was 25%, with 8% of patients requiring renal replacement therapy. Acute renal failure requiring renal replacement therapy was associated with increased mortality among both heart (odds ratio, 9.0; 95% confidence interval, 1.8-45.8) and liver transplant recipients (odds ratio, 12.1; 95% confidence interval, 3.9 37.3). This degree of acute renal failure also increased length of stay by nearly 3 weeks and charges by more than $115,000. Even among patients who did not require renal replacement, acute renal failure was strongly associated with increased mortality, length of stay, and charges. CONCLUSIONS: Acute renal failure remains a common complication of nonrenal solid organ transplantation and is associated with increased mortality, prolonged hospitalization, and significant financial costs. PMID- 15548975 TI - Living donor nephrectomy: flank incision versus anterior vertical mini-incision. AB - BACKGROUND: Currently, many centers perform laparoscopic donor nephrectomy (DN). We studied the outcome of donors and recipients following open DN using either flank incision (ODN) or mini-incision (MIDN). METHODS: Data of 196 living kidney donors were recorded prospectively. In 127 cases ODN and 69 cases MIDN were performed. RESULTS: Demographic details of donors were comparable for both groups. The left kidney was procured in 58% for ODN and in 64% for MIDN. Multiple arteries were more frequently present when MIDN (11% vs. 28%) was performed. The mean operating time was 129 min for ODN and 133 min for MIDN. Early complications occurred in 7% following ODN and in 4% following MIDN. Late complications were observed in 21% after ODN and 1% after MIDN. The mean hospital stay was significantly longer following ODN compared with MIDN (7.5 vs. 6.4 days). The primary graft function rate was 97% in both groups. One-year graft survival was 97% after ODN and 100% after MIDN. CONCLUSIONS: Results following MIDN are superior to those following ODN. Even in case of multiple renal vessels MIDN can be safely applied. In comparison with laparoscopic DN advantages of MIDN may be reduced costs, shorter operating time, and comparable cosmetic results. PMID- 15548976 TI - Investigation of pediatric renal transplant recipients with heavy proteinuria after sirolimus rescue. AB - BACKGROUND: Little is known about the incidence and evolution of proteinuria as a complication of sirolimus rescue in children. This study describes pediatric renal transplant (Tx) recipients who were treated with sirolimus and who developed heavy proteinuria. Risk factors for the development of proteinuria and its time course are explored. METHODS: Data at various time points after sirolimus introduction were abstracted from the records of children treated at the author's center. The repeated measures general linear model and the Student's paired t test were used to analyze changes in laboratory values over time. RESULTS: Of the 13 children on sirolimus, 12 developed heavy proteinuria after a mean interval of 1 month. The mean urine protein (Up)-to-creatinine (c) ratio increased from 1.1 to a peak value of 3.9 (P=0.003) at 4.6 months after the start of sirolimus. Although not statistically significant, children on no calcineurin inhibitor (CNI) had a greater increase in the Up/c than those on low-dose CNI. At last follow-up, with the use of angiotensin receptor blockers (ARB), the Up/c declined to 2.2. No predictors could be identified for the development of proteinuria. CONCLUSIONS: Heavy proteinuria is common after the use of sirolimus as rescue therapy in children with renal Tx. Whether this is attributable to a toxic effect of the sirolimus itself or to lower CNI exposure is uncertain. Early detection of proteinuria is important to enable prompt intervention. Most children have a reduction in their Up/c with the use of ARB and can therefore be continued on sirolimus. PMID- 15548977 TI - Refinement of the automated method for human islet isolation and presentation of a closed system for in vitro islet culture. AB - BACKGROUND: The procedure of human islet isolation needs further optimization and standardization. Here, we describe techniques to enhance enzymatic digestion and minimize mechanical forces during the digestion process. The isolation protocol has also been modified to meet current GMP (cGMP) standards. Moreover, the impact of donor- and process-related factors was correlated to the use of islets for clinical transplantation. METHODS: One hundred twelve standardized consecutive islet isolations were evaluated. Metyltioninklorid and indermil (topical tissue adhesive) were applied to detect leakage of collagenase injected and to repair the damaged pancreatic glands. The effects of dye and glue were evaluated in terms of islet yield, islet function using the perifusion assay, and success rate of the isolation. To analyze key factors for successful isolations, both univariate and multivariate regression analysis were performed. RESULTS: Both Metyltioninklorid and Indermil were effective to prevent leakage of enzyme solutions from the pancreatic glands. Both islet yield and success rate were higher when these tools were applied (4,516.1+/-543.0 vs. 3,447.7+/-323.5, P=0.02; 50.0% vs. 21.3%, P=0.02, respectively). No adverse effects on islet function or collagenase activity were observed. Multivariate regression analysis identified the maximal recorded amylase >100 U/L (P=0.026), BMI (P=0.03), and the use of catecholamine (P=0.04) as crucial donor-related factors. In addition, cold ischemia time (P=0.005), the dissection procedure using whole glands with duodenum (P=0.02), and the local procurement team (P=0.03) were identified as crucial isolation-related variables. CONCLUSIONS: A standardized technique of islet isolation is presented applying novel means to improve enzymatic digestion and to meet cGMP standards. PMID- 15548978 TI - High serum leptin in patients with chronic graft-versus-host disease after hematopoietic stem cell transplantation. AB - BACKGROUND: Increased serum leptin has been described after various organ transplants, with a mechanism that is still unclear. METHODS: We measured serum leptin in 60 patients before and after allogeneic (allo) or autologous (auto) stem cell transplant (SCT) and in 60 healthy controls, matched for age and body mass index (BMI). RESULTS: Serum leptin was higher in patients after SCT than before and in controls. Leptin production was higher after allo- than after auto SCT; the presence of chronic graft-versus-host disease (cGVHD) was associated with the highest values. The physiological correlation with BMI was lost in the allogeneic setting, indicating a strong influence of factors other than the nutritional status on circulating leptin. No relationship was found between serum leptin levels and time from transplant, age, cortisol, C-reactive protein, and T lymphocyte CD4-to-CD8 ratio. Among the cytokines secreted by type-1/type-2 T helper lymphocytes, only serum interferon-gamma significantly correlated with serum leptin levels. Anti-leptin blocking antibodies partially inhibited T-cell activation in mixed lymphocyte reaction, suggesting a link between leptin and T lymphocyte activation in the allo-SCT setting. CONCLUSION: Taken together, these findings suggest that increased serum leptin concentrations may contribute to T cell activation during development of cGVHD. PMID- 15548979 TI - Prospective psychosocial monitoring of living kidney donors using the Short Form 36 health survey: results at 12 months. AB - BACKGROUND: Lack of prospective psychosocial outcome studies on living kidney donors impedes identification of risk factors for poor outcome. METHODS: Psychiatric assessment of living kidney donors was performed preoperatively and at 4 and 12 months postoperatively using a semistructured interview, the Short Form (SF)-36 Health Survey, and Patient Health Questionnaire psychiatric assessment. A total of 48 of 51 consecutive donors (94%) over a 5-year period were available for follow-up and completed all assessments. RESULTS: At preoperative assessment, only 1 of the 48 donors (2%) had a Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition Axis I psychiatric disorder, but 15 (31%) developed a disorder during the 12 months, a 29% incidence. Disorders were depressive (12%), anxiety (6%), and adjustment (13%). Seven donors (15%) demonstrated a disorder at 12 months (depressive 10%, anxiety 2%, adjustment 2%). There was a corresponding decline in psychosocial function as measured by the SF-36 Mental Component Summary score; it decreased at both 4 and 12 months (P<0.01, P<0.05); for 19% of donors, this was a larger decrease than would be expected for the cohort (>2 standard error of measurement units). Scores for SF-36 scales of General Health and Vitality decreased significantly (P<0.05), as did those of Bodily Pain, indicating greater impairment from pain. Psychiatric disorder at 12 months was associated with donor psychosocial function (Mental Component Summary) and psychiatric disorder at 4 months (P<0.01), physical function (SF-36 Physical Component Summary score) at 4 and 12 months (P<0.01), and recipient psychiatric disorder at 12 months (P<0.05). CONCLUSIONS: Donors should be alerted to possible psychosocial impairment, assessed for risk factors, and monitored for at least 12 months. Treatment should be available. PMID- 15548981 TI - Rapid discontinuation of prednisone in higher-risk kidney transplant recipients. AB - Prednisone-minimization protocols have been successful in low-risk recipients. We report on the use of a protocol incorporating rapid discontinuation of prednisone in a cohort of kidney transplant recipients (n = 79) at increased immunologic risk. Our data suggests that such recipients should not be excluded from prednisone-minimization protocols. PMID- 15548980 TI - Late-onset cytomegalovirus disease in liver transplant recipients despite antiviral prophylaxis. AB - BACKGROUND: The incidence and impact of cytomegalovirus (CMV) disease that occurs despite CMV prophylaxis among liver transplant recipients have been incompletely defined. METHODS: The incidence and risk factors for CMV disease during the first posttransplant year in a cohort of liver transplant recipients who received antiviral prophylaxis with oral ganciclovir were retrospectively analyzed using Cox proportional-hazard regression models. RESULTS: CMV disease developed in 19 of 259 recipients (7% [95% confidence interval 0.04-0.11]) at a median of 4.5 months posttransplant, included syndrome (63%) or tissue-invasive disease (37%), and was independently associated with an increased risk of mortality during the first posttransplant year (hazard ratio 14 [95% confidence interval 3.8-54], P=0.0007). The incidence was higher (10/38 [26%] vs. 8/180 [4.5%], P<0.0001) in seronegative recipients (R-) of an organ from a seropositive donor (D+) compared with seropositive (R+) patients, respectively. D+R- status was the only variable significantly associated with CMV disease in multivariate analysis. CONCLUSIONS: Late CMV disease develops in a substantial proportion of D+R- recipients after prophylaxis is discontinued, is not accurately predicted by patient factors, and is associated with increased mortality. New strategies to identify D+R- patients at risk and to reduce the incidence and impact of late CMV disease in this group are warranted. PMID- 15548982 TI - Low-dose cyclosporine with mycophenolate mofetil induces similar calcineurin activity and cytokine inhibition as does standard-dose cyclosporine in stable renal allografts. AB - One strategy to minimize nephrotoxicity in maintenance immunosuppression in renal transplantation is reduction of cyclosporine (CsA) with addition of mycophenolate mofetil (MMF). This approach seems safe, but concern exists about whether it yields adequate immunosuppression in the long term. Thus, we investigated the pharmacodynamic response to CsA in stable renal allografts treated with standard CsA (n = 17, CsA-C0h > or = 125 ng/mL) and low CsA plus MMF (n = 18 CsA-C0h <100 ng/mL). Patients treated with MMF without CsA (n = 13) and healthy subjects (n = 7) were used as controls. We observed that inhibition of calcineurin (CN) activity in peripheral blood mononuclear cells (PBMC), as well as interleukin (IL)-2 and interferon (IFN)-gamma production were similar in Standard-CsA and Low CsA+MMF groups. Moreover, addition of MMF to a low CsA dose regime improved the correlation between CsA-C2h and both CN activity and IL-2 production. Thus, our results suggest that MMF could be synergistic with the pharmacodynamic effect of low CsA in maintenance immunosuppression. PMID- 15548983 TI - Hematochezia and congenital extrahepatic portocaval shunt with absent portal vein: successful treatment by liver transplantation. PMID- 15548984 TI - Delirium in a renal transplant recipient associated with BK virus in the cerebrospinal fluid. PMID- 15548986 TI - Adenovirus-mediated gene transfer to the rat heart graft. PMID- 15548990 TI - Hematopoietic stem cell transplantation. PMID- 15548991 TI - Umbilical cord blood transplantation. AB - PURPOSE OF REVIEW: Familiar and unrelated umbilical cord blood is an appealing alternative source of hematopoietic stem cells patients undergoing transplantation for a wide variety of diseases. In the unrelated donor transplant setting, shorter time to transplant, which is particularly relevant to patients requiring urgent transplantation, and tolerance of 1-2 human leukocyte antigen mismatch, which increases the chance of finding a suitable donor, are evident advantages over bone marrow transplantation. The speed of engraftment is slower after cord blood transplantation but it is counterbalanced by a lower incidence of severe graft-versus-host disease. Cell dose and human leukocyte antigen are major factors influencing outcome after umbilical cord blood transplantation. RECENT FINDINGS: Unrelated donor cord blood transplantation is considered an acceptable option to bone marrow for pediatric transplantation, and recent data in adults point the same way. SUMMARY: This review describes the recent clinical results of cord blood transplantation and discusses developing research strategies aimed at optimizing this kind of transplantation. PMID- 15548992 TI - HLA matching in allogeneic stem cell transplantation. AB - PURPOSE OF REVIEW: The success of unrelated hematopoietic cell transplantation (HCT) is influenced by the degree of HLA compatibility between the donor and patient. The goal of this review is to summarize new findings in the field of immunogenetics and HCT from unrelated donors using myeloablative conditioning regimens. RECENT FINDINGS: Molecular typing methods can discriminate unique alleles encoded by HLA class I and II genes. Incompatibility of donor-recipient HLA alleles increases posttransplant complications including graft rejection, acute and chronic graft-versus-host disease, and mortality. These posttransplant risks increase with increasing numbers of HLA mismatches. The identification of permissible HLA mismatches may be aided by the use of functional assays. Nongenetic factors, including the stage of disease at the time of transplantation, may influence the effect of HLA mismatching on survival. SUMMARY: HLA alleles are functionally relevant. Unrelated HCT can be optimized by comprehensive and precise donor-recipient allele matching. For patients with high risk diseases who lack matched donors, use of donors with a single HLA mismatch may permit early treatment before disease progression. PMID- 15548993 TI - Recent advances in hematopoietic stem cell biology. AB - PURPOSE OF REVIEW: Exciting advances have been made in the field of hematopoietic stem cell biology during the past year. This review summarizes recent progress in the identification, culture, and in vivo tracking of hematopoietic stem cells. RECENT FINDINGS: The roles of Wnt and Notch proteins in regulating stem cell renewal in the microenvironment, and how these molecules can be exploited in ex vivo stem cell culture, are reviewed. The importance of identification of stem cells using functional as well as phenotypic markers is discussed. The novel field of nanotechnology is then discussed in the context of stem cell tracking in vivo. This review concludes with a section on the unexpected potential of bone marrow-derived stem cells to contribute to the repair of damaged tissues. The contribution of cell fusion to explain the latter phenomenon is discussed. SUMMARY: Because of exciting discoveries made recently in the field of stem cell biology, researchers now have improved tools to define novel populations of stem cells, examine them ex vivo using conditions that promote self-renewal, track them into recipients, and determine whether they can contribute to the repair of damaged tissues. These discoveries will significantly advance the field of stem cell transplantation. PMID- 15548994 TI - Stem cell therapy for cardiac diseases. AB - PURPOSE OF REVIEW: This review examines the knowledge that researchers have gained during the past year regarding some fundamental questions about stem cell therapy. These questions concern patient selection and safety, the optimal type of stem cells, the best route for their delivery, the fate of the transplanted cells, and the mechanism by which this therapy works. RECENT FINDINGS: So far, candidates for cardiac stem cell therapy have been limited to patients with acute myocardial infarction and chronic ischemic heart failure. Currently, bone marrow stem cells seem to be the most attractive cell type for these patients. The cells may be delivered by means of direct surgical injection, intracoronary infusion, retrograde venous infusion, and transendocardial injection. Stem cells may directly increase cardiac contractility or passively limit infarct expansion and remodeling. This therapy is generally well tolerated, but the potential for accelerated atherogenesis remains a concern. Eventually, cell therapy may be combined with gene therapy to treat ischemic myocardium. SUMMARY: Stem cell therapy for cardiac disease is a rapidly evolving field. Most of the evidence accumulated so far, including preclinical and clinical findings, confirms the potential of this novel therapy. However, most of the fundamental knowledge needed to guide the application of stem cell therapy in cardiac disease is still lacking. PMID- 15548995 TI - Interleukin-18: recent advances. AB - PURPOSE OF REVIEW: Interleukin-18 (IL-18) has potent immunomodulatory effects. It is the only cytokine with a unique capacity to induce T helper 1 or T helper 2 polarization, depending on the immunologic context. Serum levels of IL-18 are increased in many human diseases and it has been implicated in the pathogenesis of several immune-mediated processes. Some of the recent key advances in the immunobiology of IL-18 are discussed in this review. RECENT FINDINGS: Recent data from several laboratories have shed light on the structure of IL-18; the signaling cascades that are initiated; and its role on modulating T cells, dendritic cells, and natural killer cell function. Several new reports have expanded and delineated the role of IL-18 in a multitude of diseases, but only recent advances in the role of IL-18 in three disease processes (acute graft versus-host disease, insulin-dependent diabetes, and sepsis), where it appears to play paradoxic roles are discussed. SUMMARY: Although emerging data shed more light on the complex role of IL-18 in immune reactions, they also pose more questions. Given the pleiotropic, complex, and at times paradoxic effects of IL 18 in various disease processes, better understanding of its immunobiology might lead to the development of IL-18 and/or its antagonists as therapeutic agents against immune-mediated diseases. PMID- 15548996 TI - Mantle cell lymphoma. AB - PURPOSE OF REVIEW: Mantle cell lymphoma is the B-cell lymphoma with the worst prognosis. Until now, no standard treatment has resulted in cure. Improvements in understanding of the disease are needed to advance therapeutic efforts. RECENT FINDINGS: Pathology and immunohistochemistry can identify the subset of patients with the worse prognosis. New data suggest that at least a subset of mantle cell lymphoma cases have undergone some form of antigene selection, and particular types of Ig gene rearrangement seem to give a better prognosis. The cell cycle, the ATM, gene and the nuclear factor kappaB pathways are the main targets of the genetic abnormalities occurring in mantle cell lymphoma: new genomic and expression data have been recently published. Unfortunately, this progress has not yet brought any major improvements in therapeutic approaches, which still remain highly unsatisfactory. Autologous and allogenic bone marrow transplantations appear to be the only current treatments that might improve the outcome of patients with PMCL. New additional treatment modalities are currently under investigation. SUMMARY: This review summarizes all the most recent data published on the biology and treatment of mantle cell lymphoma. PMID- 15548997 TI - Antibody-targeted therapy: a paradigm of innovative treatment strategies in indolent and aggressive B-cell non-Hodgkin lymphoma. AB - PURPOSE OF REVIEW: This review outlines the principles of treatment of indolent and aggressive B-cell lymphoma based on current knowledge on the classification of hematologic malignancies and the rationale to implement new antibody-targeted immunotherapeutic approaches. RECENT FINDINGS: An update is provided on the use of antibody-targeted therapies in clinical trials, with emphasis on new, emerging strategies of immunotherapy in B-cell non-Hodgkin lymphoma. SUMMARY: The success of immune-mediated therapies has encouraged studies on antibody-targeted therapy in B-cell non-Hodgkin lymphoma. Promising new approaches combine classical dose intense chemotherapy with "tumor-specific" antibody targeting during several phases of the disease. The safety and efficacy of anti-CD20 in the treatment of indolent and aggressive B-cell non-Hodgkin lymphoma at any stage of disease, either as a single agent or as part of multimodality regimes, as an unconjugated antibody or as radioimmunoconjugate have changed dramatically our treatment strategies. Increasing insights into basic molecular biology and immunology of B cell non-Hodgkin lymphoma may identify subgroups of patients categorized in current classification systems who may benefit from tailored approaches with new modality antibody-targeted therapy in near future. PMID- 15548998 TI - T cell acute lymphoblastic leukemia/lymphoma: a human cancer commonly associated with aberrant NOTCH1 signaling. AB - PURPOSE OF REVIEW: Although constitutively activated forms of the NOTCH1 receptor are potent inducers of T cell acute lymphoblastic leukemia/lymphoma when expressed in the bone marrow stem cells of mice, the known involvement of NOTCH1 in human T cell acute lymphoblastic leukemia/lymphoma has been restricted to very rare tumors associated with a (7;9) chromosomal translocation involving the NOTCH1 gene. This picture has changed dramatically in the past year with the discovery of frequent mutations involving NOTCH1 in human T cell acute lymphoblastic leukemia/lymphoma. RECENT FINDINGS: NOTCH1 point mutations, insertions, and deletions producing aberrant increases in NOTCH1 signaling are frequently present in both childhood and adult T cell acute lymphoblastic leukemia/lymphoma and are detected in tumors from all major molecular subtypes. These observations are particularly important in the light of experiments using human and murine T cell acute lymphoblastic leukemia/lymphoma cell lines indicating that NOTCH1 signals are required for sustained growth and, in a subset of lines, survival. This inference is based in part on experiments conducted with small molecule inhibitors of gamma-secretase, a protease required for normal NOTCH signal transduction and the activity of the mutated forms of NOTCH1 found commonly in human T cell acute lymphoblastic leukemia/lymphoma. SUMMARY: These findings support a central role for aberrant NOTCH signaling in the pathogenesis of human T cell acute lymphoblastic leukemia/lymphoma, and they provide a rationale for trials of NOTCH inhibitors, such as gamma-secretase antagonists, in this aggressive human malignancy. PMID- 15549000 TI - Bibliography. Current world literature. Hematopoietic stem cell transplantation. PMID- 15548999 TI - Pharmacogenetics of acute lymphoblastic leukemia. AB - PURPOSE OF REVIEW: The outcome in children with acute lymphoblastic leukemia has improved significantly over the past four decades. Current therapy results in event-free survival exceeding 80% for most patients. The development of risk adapted therapy based on characteristics of the child (age), leukemia (leukocyte count, acquired genetic characteristics) and early response to therapy allows dose intensification for children with higher-risk disease. Much less attention has been given to the role of host variability (pharmacogenetic polymorphism) in determining outcome. This review discusses literature reports in this area and describes some of the challenges facing the field as it moves forward. RECENT FINDINGS: Polymorphisms in many different metabolic pathways have been demonstrated in single gene studies to influence the outcome of acute lymphoblastic leukemia. Challenges arise in establishing the generalizability of observations and interpreting complex gene-gene interactions in multigene pathways. Recent studies also illustrate the importance of correlation of clinical associations with biological mechanisms. SUMMARY: Despite significant progress in the treatment of childhood acute lymphoblastic leukemia, therapy is still unsuccessful in 20% of patients. Further knowledge of and insight into the role of host genetic polymorphisms will improve the results by integrating pharmacodynamic and pharmacogenomic studies in individualizing therapy for children with acute lymphoblastic leukemia. PMID- 15549001 TI - Gender: a relevant analytic key to palliative care? PMID- 15549002 TI - Identifying the palliative care role of the nurse working in community hospitals: an exploratory study. AB - AIM: to ascertain registered nurses' and GPs' perceptions of and influences on the nurse's role in providing palliative care in community hospitals. DESIGN: qualitative exploratory/descriptive study. SETTING: two community hospital sites within Perth and Kinross, Scotland. METHOD: taped semi-structured interviews with four registered nurses and two GPs. Participants' perceptions were sought on the roles of nursing and medical professionals and of factors facilitating and hindering the provision of palliative care within the community hospital setting. FINDINGS: six key themes were identified through thematic analysis of the information: communication, teamwork and relationships, holistic care, resources, culture, professional role. CONCLUSION: the themes were inter-related and gave valuable insight into the multidimensional nature and significance of the nurse's role in palliative care in the community hospital setting. PMID- 15549003 TI - Grief in North America, a death-denying society. PMID- 15549004 TI - The impact of unrelieved patient suffering on palliative care nurses. AB - AIM: to describe the impact of unrelieved patient suffering on nurses working with palliative care patients. DESIGN: This was a qualitative descriptive design using semi-structured interviews. SAMPLE: nine experienced palliative care nurses were interviewed. RESULTS: nurses acknowledged that the term 'suffering' generally was not used in the workplace. The nurses identified that only a small group of patients died with suffering that could be classified as 'unrelieved' but that the impact of these patients' suffering on themselves was enormous. Nurses describe the impact in terms of perceptions of suffering (difficult situation), feelings (helplessness, distress, feelings of failure), bearing the burden (alcohol consumption, headaches) and effects on their relationship with family. The nurses identified several factors that increased the personal impact of unrelieved patient suffering. The most important strategy for ameliorating the impact of unrelieved patient suffering was informal support from work colleagues. CONCLUSION: the nurses' stories indicate that the personal impact of unrelieved patient suffering could be reduced through acknowledgement of this suffering and better formal and informal support mechanisms. PMID- 15549005 TI - The development of an interdisciplinary outpatient clinic in specialist palliative care. AB - The interdisciplinary team approach is essential in the assessment and management of the palliative care of patients and their families. An innovative approach has been developed to allow the interdisciplinary team to see patients and their families in an outpatient clinic setting. This has allowed an improved assessment and has demonstrated to patients that a wider approach is undertaken. These interdisciplinary outpatient clinics have been audited and appear to be effective, and acceptable to patients and families. PMID- 15549006 TI - Assessing palliative care outcomes for people with motor neurone disease living at home. AB - A number of palliative care outcome measures are used to facilitate the provision of palliative care. This short article reports the use of one palliative care outcome measure, the Palliative care Outcome Scale, with people with motor neurone disease (MND) living at home. The outcome measure was generally viewed positively by those involved. However, problems with this scale itself point to the need for a specialized instrument to assess palliative care outcomes for people living with MND. PMID- 15549007 TI - Handling unexpected distress on the telephone: the development of interdisciplinary training. AB - St Nicholas's Hospice recognized that its non-clinical staff needed support and guidance regarding the management of particularly difficult telephone conversations. It is understandable that patients and family members who are experiencing extreme uncertainty, fear and worry might express anger and despair, however, non-clinical staff were troubled by having to handle such raw emotions on the telephone. Despite palliative care excelling at counselling skills and training in breaking bad news, no material or courses exist to help these staff handle unanticipated distress on the telephone. A training session was established and, unexpectedly, many clinical staff enrolled. Although a formal in depth evaluation did not take place, participant feedback suggested it was equally beneficial to non-clinical and clinical staff. A key element of the apparent success was the interprofessional, interagency, interpersonal sharing and exchange of ideas. PMID- 15549009 TI - Nurses to take over GPs' emergency call service. PMID- 15549010 TI - Management of patients who have relapses in multiple sclerosis. AB - Multiple sclerosis (MS) is a chronic neurological condition, which disrupts the typical functions and daily activities of those affected. Approximately 80% of those diagnosed will follow a relapsing/remitting disease course at diagnosis. Patients with relapsing/remitting MS can expect to have episodes of relapse or an exacerbation of the disease and its variety of symptoms. Management of such episodes has always been open to the preferences of the healthcare professionals involved. The aim of this article is to review the evidence that exists to support current practice in the management of patients during a relapse and to evaluate how this evidence can be incorporated into patient care. The article begins by giving an overview of MS, and continues by determining what a relapse is and discussing the speculated triggers of relapse. Methods of treating relapse are then explored. It concludes by highlighting the strongest evidence-based method for treating relapses, and by emphasizing the need for further research into the pharmaceutical and non-pharmaceutical management of MS relapses. PMID- 15549012 TI - A nurse-led, patient-centred mitoxantrone service in neurology. AB - Multiple sclerosis (MS) is an autoimmune disease of the central nervous system for which there is no known cure. There are licensed treatments available to influence the disease course, but for a small group of patients, progression of the disease may continue at an alarming rate. Mitoxantrone, a cytotoxic agent normally used in the treatment of cancer, has been shown to reduce disease activity in patients who have this more aggressive form of MS. However, the immunosuppressive action and side-effect profile of mitoxantrone mean that this drug should be used with caution. This article describes a MS nurse-led initiative where an evidence-based, integrated care pathway was designed to ensure a safe, high-quality service provision for this select patient group. PMID- 15549013 TI - Nurse with alcohol problem who failed in her care of patients. PMID- 15549011 TI - The management of pain in multiple sclerosis: a care-study approach. AB - Pain is a commonly reported symptom of multiple sclerosis (MS). However, the literature surrounding pain management for people with MS appears limited. This article describes how pain was managed for one patient. It suggests that the MS nurse is pivotal in ensuring that patients receive adequate pain control. The MS nurse can coordinate timely assessment, monitor effectiveness and side effects linked with a pain management plan, and provide education and support to patients in order to allow them to take responsibility for their own pain management. PMID- 15549014 TI - Decontamination of medical devices: legislation and compliance to practice. AB - The amount of legislation and guidelines published on decontamination reflects the current attitude of politicians and the general public to the desire to reduce the risk of infection. Incidents, such as bovine spongiform encephalopathy (BSE) in cattle with the subsequent risk of transmission of variant Creutzfeldt Jakob disease (VCJD), require greater compliance by healthcare workers to the guidance both in the hospital and the wider community. The change in care provision and the demands of government in training and education. This article will look at recent government guidance and the implications it has for clinical practice. PMID- 15549015 TI - Not for resuscitation instructions: the law for children in the UK. AB - CASE SCENARIO: David Glass, a boy of 13 years, was severely disabled with only a limited life span. His mother wished him to receive whatever medical treatment was necessary to prolong his life. Following an incident in which the hospital gave the child diamorphine against the mother's wishes, family members resuscitated the child and prevented him from dying. There was a complete breakdown of trust between the family and the hospital. His mother, Ms Glass, sought a declaration as to the course doctors in the hospital should take if the boy were admitted for emergency treatment and disagreements arose as to the treatment to be given to or withheld from the child. The judge refused the mother's application for judicial review and she appealed to the Court of Appeal (R v. Portsmouth Hospitals NHS Trust ex p. Glass [1999]). PMID- 15549016 TI - The introduction of a "trial without catheter" model in primary care. AB - Action research was used to compare and evaluate the current nursing practice of urinary catheter removal. The aim of this study was to plan, implement and evaluate a trial without catheter (TWOC) model in primary care and compare it with the clinical procedure being performed in the secondary care setting. Two sample groups were used; the first sample consisted of 10 patients who had undergone a TWOC in hospital, and the second consisted of 10 patients who had a TWOC in the community. An interview with the urology nurse specialist was also undertaken to gain further qualitative data. Statistics were analysed using an unpaired t-test. Results showed that 80% of hospital patients and all community patients stated that given the choice they would prefer to have their TWOC at home. Infection rates were monitored and 40% of hospital TWOC patients developed a urine infection, whereas no infection was reported in the community group. From this small study, it was evident that there were major benefits to providing a TWOC service within the community setting. PMID- 15549017 TI - Traumatic injuries to the head and spine 2: nursing considerations. AB - Trauma to the central nervous system can have devastating consequences for both the person who sustained the injury and his/her family/loved ones. This article first discusses pathophysiology in relation to altered cerebral haemodynamics and changes that occur after spinal injury. Following on from the underpinning theoretical perspectives, the article reviews the nursing care and management strategies required by patients who have sustained either a traumatic head injury or acute spinal injury, with the aim of controlling secondary injury, which is preventable. This ensures the patient will have the best possible prognosis and outcome. PMID- 15549018 TI - JOBST opaque compression hosiery in the management of venous disease. AB - Prevention of leg ulceration is a simple process involving assessment, accurate measurement of the leg and application of appropriate compression hosiery along with appropriate care of the skin. Persuasion of at-risk individuals to be aware of and follow prevention options needs to be incorporated in the ongoing health promotion activities of the practitioner (Price and Harding, 1996). This product focus examines the potential contributions of the new JOBST Opaque gradient compression hosiery (with the innovative JOBST soft care yarn) in prevention and management of venous disease and deep vein thrombosis, without sacrificing comfort and style in order to promote patient concordance. PMID- 15549019 TI - How to survive nursing school: a practical guide. PMID- 15549020 TI - Acinetobacter baumannii infections among patients at military medical facilities treating injured U.S. service members, 2002-2004. AB - Acinetobacter baumannii is a well known but relatively uncommon cause of health care--associated infections. Because the organism has developed substantial antimicrobial resistance, treatment of infections attributed to A. baumannii has become increasingly difficult This report describes an increasing number of A. baumannii bloodstream infections in patients at military medical facilities in which service members injured in the Iraq/Kuwait region during Operation Iraqi Freedom (OIF) and in Afghanistan during Operation Enduring Freedom (OEF) were treated. The number of these infections and their resistance to multiple antimicrobial agents underscore 1) the importance of infection control during treatment in combat and health-care settings and 2) the need to develop new antimicrobial drugs to treat these infections. PMID- 15549021 TI - Prevalence of overweight and obesity among adults with diagnosed diabetes--United States, 1988-1994 and 1999-2002. AB - Obesity in persons with diabetes is associated with poorer control of blood glucose levels, blood pressure, and cholesterol, placing persons with diabetes at higher risk for both cardiovascular and microvascular disease. Conversely, intentional weight loss is associated with reduced mortality among overweight persons with diabetes. CDC analyzed the prevalence of overweight and obesity among U.S. adults aged >/=20 years with previously diagnosed diabetes by using data from two surveys: the Third National Health and Nutrition Examination Survey (NHANES III), 1988-1994, and NHANES 1999-2002. This report summarizes the results of that analysis, which indicated that most adults with diagnosed diabetes were overweight or obese. During 1999-2002, the prevalence of overweight or obesity was 85.2%, and the prevalence of obesity was 54.8%. Encouraging patients to achieve and maintain a healthy weight should be a priority for all diabetes-care programs. PMID- 15549022 TI - Prevalence of visual impairment and selected eye diseases among persons aged >/=50 years with and without diabetes--United States, 2002. AB - Visual impairment and blindness affect an estimated 3.4 million U.S. adults aged >/=40 years. The leading causes of visual impairment and blindness are diabetic retinopathy and age-related eye diseases (e.g., cataracts, macular degeneration, and glaucoma). Diabetes affects approximately 18 million U.S. adults, of whom an estimated 30% have undiagnosed diabetes, and imposes an increased risk for eye disease. To characterize the prevalence of visual impairment and selected eye diseases (i.e., diabetic retinopathy, cataracts, macular degeneration, and glaucoma) among persons aged >50 years with and without diabetes, CDC analyzed data from the 2002 National Health Interview Survey (NHIS). This report summarizes the findings of that analysis, which identified a substantially higher prevalence of visual impairment and eye disease among those with diabetes compared with those without diabetes. Measures are needed to increase comprehensive eye examinations, especially among adults at high risk for blindness and visual impairment (e.g., persons aged >/=65 years and those with diabetes). PMID- 15549023 TI - West Nile virus activity--United States, November 9-16, 2004. AB - During November 9-16, a total of 31 cases of human West Nile virus (WNV) illness were reported from eight states (Arizona, Georgia, Maryland, Michigan, Missouri, New York, Pennsylvania, and Texas). PMID- 15549039 TI - Pain during childbirth. PMID- 15549050 TI - Benign focal liver lesions: discrimination from malignant mimickers. AB - Focal lesions of the liver often have various imaging characteristics which may be interpreted as either benign or malignant. Understanding the underlying pathophysiology of these liver lesions may lead to characteristic imaging manifestations, which direct the radiologist to the diagnosis. Benign lesions include congenital hepatic cyst, autosomal dominant polycystic disease, hemangioma, focal nodular hyperplasia (FNH), hepatic adenoma, inflammatory pseudotumor, peliosis hepatis, focal fatty infiltration, hamartoma, and infectious processes such as hepatic abscess, echinococcal cyst, and candidiasis. Characteristic imaging features, clinical symptoms, and treatment/prognosis will be discussed. Emphasis will be placed on key reliable features of each disease to develop a method of discriminating these lesions from other benign and malignant disorders. PMID- 15549051 TI - Malignant cystic and necrotic liver lesions: a pattern approach to discrimination. AB - Malignant primary and metastatic lesions of the liver have a myriad of imaging appearances. Discriminating between the various lesions requires an understanding of the underlying pathophysiology and imaging characteristics that lead to their malignant appearances. A pattern approach of recognition by imaging with understanding of why particular lesions behave the way they do on sonography, CT, MRI, and nuclear scintigraphy can be developed. A broad selection of malignant cystic liver lesions will be displayed in a multimodality pictorial style including hepatocellular carcinoma, cholangiocarcinoma, cystic and necrotic hepatic metastases, biliary cystadenoma/cystadenocarcinoma, epitheloid hemangioendothelioma, hepatoblastoma, and cystic sarcomas. Characteristic imaging features, clinical symptoms, and prognosis will be discussed. Emphasis will be placed on consistent features of these malignant lesions that help to discriminate them from other hepatic disorders. PMID- 15549052 TI - Endovascular repair of thoracic aortic disease: what we have learned. PMID- 15549053 TI - Rapid dopamine signaling in the nucleus accumbens during contingent and noncontingent cocaine administration. AB - Cocaine acts as a reinforcer through its pharmacological effects on brain monoaminergic systems, which, through repeated pairings with environmental stimuli, lead to the development of conditioned effects of the drug. Both the pharmacological and conditioned aspects of cocaine are implicated in several facets of acquisition and maintenance of addiction, including drug craving. Here, we compare the effects of contingent (response dependent) and noncontingent (response independent) cocaine on rapid dopaminergic signaling in the core of the nucleus accumbens. Dopamine was monitored using fast-scan cyclic voltammetry. Noncontingent cocaine administered to both naive and animals with a history of self-administration resulted in a profound increase in the frequency of transient dopamine release events that are not time-locked to any specific environmental stimuli. Pharmacological effects were detectable approximately 40 s after cocaine administration. In contrast, when animals where allowed to self-administer cocaine on an FR-1 schedule, dopamine transients (69+/-12 nM) were consistently observed time-locked to each reinforced response (peaking approximately 1.5 s after response completion). Importantly, no pharmacological effect of cocaine was observed within the 10 s following noncontingent cocaine administration, indicating that dopamine signals time-locked to the reinforced response are a result of the pairing of the operant behavior, the drug-associated cues, and cocaine. These data demonstrate that this pharmacological action of cocaine occurs for an extended period following either contingent or noncontingent administration, but is distinct from those dopamine transients that are time locked to each lever-press in self-administering animals. PMID- 15549055 TI - A chance for growth. PMID- 15549056 TI - Clean, green conferencing. PMID- 15549057 TI - Campaign to fight malaria hit by surge in demand for medicine. PMID- 15549058 TI - Grade expectations for German research institutes. PMID- 15549054 TI - Pharmacological and anatomical evidence for an interaction between mGluR5- and GABA(A) alpha1-containing receptors in the discriminative stimulus effects of ethanol. AB - The discriminative stimulus properties of ethanol are mediated in part by positive modulation of GABA(A) receptors. Recent evidence indicates that metabotropic glutamate receptor subtype 5 (mGluR5) activity can influence GABA(A) receptor function. Therefore, the purpose of this work was to examine the potential involvement of mGluR5 in the discriminative stimulus effects of ethanol. In rats trained to discriminate ethanol (1 g/kg, intragastric gavage (i.g.)) from water, 2-methyl-6-(phenylethyl)-pyridine (MPEP) (1-50 mg/kg, i.p.) a selective noncompetitive antagonist of the mGlu5 receptor did not produce ethanol like stimulus properties. However, pretreatment with MPEP (30 mg/kg) reduced the stimulus properties of ethanol as indicated by significant reductions in ethanol appropriate responding, specifically at 0.5 and 1 g/kg ethanol, and a failure of ethanol test doses (1 and 2 g/kg) to fully substitute for the ethanol training dose. To test whether mGluR5 antagonism altered the GABA(A) receptor component of the ethanol stimulus, the ability of MPEP to modulate pentobarbital and diazepam substitution for ethanol was assessed. Pentobarbital substitution (1-10 mg/kg, i.p.) for ethanol was not altered by MPEP pretreatment. However, MPEP pretreatment inhibited the ethanol-like stimulus properties of diazepam (5 mg/kg, i.p.). To examine a potential anatomical basis for these pharmacological findings, expression patterns of mGluR5- and benzodiazepine-sensitive GABA(A) alpha1-containing receptors were examined by dual-label fluorescent immunohistochemistry with visualization by confocal microscopy. Results indicated that mGluR5- and GABA(A) alpha1-containing receptors were both coexpressed in limbic brain regions and colocalized on the same cells in specific brain regions including the amygdala, hippocampus, globus pallidus, and ventral pallidum. Together, these findings suggest an interaction between mGluR5- and benzodiazepine-sensitive GABA(A) receptors in mediating ethanol discrimination. PMID- 15549059 TI - Europe's stem-cell workers pull together. PMID- 15549060 TI - Grant-transfer plan paves the way for European mobility. PMID- 15549061 TI - WHO calls for vaccine boost to prepare for flu pandemic. PMID- 15549062 TI - Stalemate over fusion project threatens to provoke split. PMID- 15549063 TI - Molecular biology enjoys double celebration. PMID- 15549064 TI - Britain to combat conflicts of interest in drug regulators. PMID- 15549065 TI - Unanimous vote approves tweak to smallpox genome. PMID- 15549068 TI - Benoit Mandelbrot: father of fractals. PMID- 15549069 TI - Emissions trading: the carbon game. PMID- 15549070 TI - Bible study led Newton to scientific discoveries. PMID- 15549071 TI - Public participation: let the people pick projects. PMID- 15549077 TI - Going against the flow. PMID- 15549072 TI - Let's be sensible about public participation. PMID- 15549078 TI - Neurobiology: at the root of brain cancer. PMID- 15549079 TI - Chemistry: towards tomorrow's catalysts. PMID- 15549080 TI - Evolutionary biology: butterfly mimics of ants. PMID- 15549082 TI - Nonlinear optics: disorder is the new order. PMID- 15549083 TI - Structural biology: ion pump in the movies. PMID- 15549085 TI - Historical phenology: grape ripening as a past climate indicator. AB - French records of grape-harvest dates in Burgundy were used to reconstruct spring summer temperatures from 1370 to 2003 using a process-based phenology model developed for the Pinot Noir grape. Our results reveal that temperatures as high as those reached in the 1990s have occurred several times in Burgundy since 1370. However, the summer of 2003 appears to have been extraordinary, with temperatures that were probably higher than in any other year since 1370. PMID- 15549086 TI - Atmospheric science: early peak in Antarctic oscillation index. AB - The principal extratropical atmospheric circulation mode in the Southern Hemisphere, the Antarctic oscillation (or Southern Hemisphere annular mode), represents fluctuations in the strength of the circumpolar vortex and has shown a trend towards a positive index in austral summer in recent decades, which has been linked to stratospheric ozone depletion and to increased atmospheric greenhouse-gas concentrations. Here we reconstruct the austral summer (December January) Antarctic oscillation index from sea-level pressure measurements over the twentieth century and find that large positive values, and positive trends of a similar magnitude to those of past decades, also occurred around 1960, and that strong negative trends occurred afterwards. This positive Antarctic oscillation index and large positive trend during a period before ozone-depleting chemicals were released into the atmosphere and before marked anthropogenic warming, together with the later negative trend, indicate that natural forcing factors or internal mechanisms in the climate system must also strongly influence the state of the Antarctic oscillation. PMID- 15549087 TI - Climate: large-scale warming is not urban. AB - Controversy has persisted over the influence of urban warming on reported large scale surface-air temperature trends. Urban heat islands occur mainly at night and are reduced in windy conditions. Here we show that, globally, temperatures over land have risen as much on windy nights as on calm nights, indicating that the observed overall warming is not a consequence of urban development. PMID- 15549090 TI - Targeted cancer therapy. AB - Disruption of the normal regulation of cell-cycle progression and division lies at the heart of the events leading to cancer. Complex networks of regulatory factors, the tumour microenvironment and stress signals, such as those resulting from damaged DNA, dictate whether cancer cells proliferate or die. Recent progress in understanding the molecular changes that underlie cancer development offer the prospect of specifically targeting malfunctioning molecules and pathways to achieve more effective and rational cancer therapy. PMID- 15549091 TI - G1 cell-cycle control and cancer. AB - Before replicating DNA during their reproductive cycle, our cells enter a phase called G1 during which they interpret a flood of signals that influence cell division and cell fate. Mistakes in this process lead to cancer. An increasingly complex and coherent view of G1 signalling networks, which coordinate cell growth, proliferation, stress management and survival, is helping to define the roots of malignancies and shows promise for the development of better cancer therapies. PMID- 15549092 TI - Intrinsic tumour suppression. AB - Mutations that drive uncontrolled cell-cycle progression are requisite events in tumorigenesis. But evolution has installed in the proliferative programmes of mammalian cells a variety of innate tumour-suppressive mechanisms that trigger apoptosis or senescence, should proliferation become aberrant. These contingent processes rely on a series of sensors and transducers that act in a coordinated network to target the machinery responsible for apoptosis and cell-cycle arrest at different points. Although oncogenic mutations that disable such networks can have profound and varied effects on tumour evolution, they may leave intact latent tumour-suppressive potential that can be harnessed therapeutically. PMID- 15549093 TI - Cell-cycle checkpoints and cancer. AB - All life on earth must cope with constant exposure to DNA-damaging agents such as the Sun's radiation. Highly conserved DNA-repair and cell-cycle checkpoint pathways allow cells to deal with both endogenous and exogenous sources of DNA damage. How much an individual is exposed to these agents and how their cells respond to DNA damage are critical determinants of whether that individual will develop cancer. These cellular responses are also important for determining toxicities and responses to current cancer therapies, most of which target the DNA. PMID- 15549094 TI - Tissue repair and stem cell renewal in carcinogenesis. AB - Cancer is increasingly being viewed as a stem cell disease, both in its propagation by a minority of cells with stem-cell-like properties and in its possible derivation from normal tissue stem cells. But stem cell activity is tightly controlled, raising the question of how normal regulation might be subverted in carcinogenesis. The long-known association between cancer and chronic tissue injury, and the more recently appreciated roles of Hedgehog and Wnt signalling pathways in tissue regeneration, stem cell renewal and cancer growth together suggest that carcinogenesis proceeds by misappropriating homeostatic mechanisms that govern tissue repair and stem cell self-renewal. PMID- 15549095 TI - Stromal fibroblasts in cancer initiation and progression. AB - It is widely accepted that the development of carcinoma--the most common form of human cancer--is due to the accumulation of somatic mutations in epithelial cells. The behaviour of carcinomas is also influenced by the tumour microenvironment, which includes extracellular matrix, blood vasculature, inflammatory cells and fibroblasts. Recent studies reveal that fibroblasts have a more profound influence on the development and progression of carcinomas than was previously appreciated. These new findings have important therapeutic implications. PMID- 15549096 TI - Aneuploidy and cancer. AB - In contrast to normal cells, aneuploidy--alterations in the number of chromosomes -is consistently observed in virtually all cancers. A growing body of evidence suggests that aneuploidy is often caused by a particular type of genetic instability, called chromosomal instability, which may reflect defects in mitotic segregation in cancer cells. A better understanding of the molecular mechanisms leading to aneuploidy holds promise for the development of cancer drugs that target this process. PMID- 15549097 TI - Endurance running and the evolution of Homo. AB - Striding bipedalism is a key derived behaviour of hominids that possibly originated soon after the divergence of the chimpanzee and human lineages. Although bipedal gaits include walking and running, running is generally considered to have played no major role in human evolution because humans, like apes, are poor sprinters compared to most quadrupeds. Here we assess how well humans perform at sustained long-distance running, and review the physiological and anatomical bases of endurance running capabilities in humans and other mammals. Judged by several criteria, humans perform remarkably well at endurance running, thanks to a diverse array of features, many of which leave traces in the skeleton. The fossil evidence of these features suggests that endurance running is a derived capability of the genus Homo, originating about 2 million years ago, and may have been instrumental in the evolution of the human body form. PMID- 15549098 TI - Molecular hydrogen beyond the optical edge of an isolated spiral galaxy. AB - Knowledge about the outermost portions of galaxies is limited owing to the small amount of light coming from them. It is known that in many cases atomic hydrogen (H I) extends well beyond the optical radius. In the centres of galaxies, however, molecular hydrogen (H2) usually dominates by a large factor, raising the question of whether H2 is also abundant in the outer regions. Here we report the detection of emission from carbon monoxide (CO), the most abundant tracer of H2, beyond the optical radius of the nearby galaxy NGC 4414. The host molecular clouds probably formed in the regions of relatively high H i column density and in the absence of spiral density waves. The relative strength of the lines from the two lowest rotational levels indicates that both the temperature and density of the H2 are quite low compared to conditions closer to the centre. The inferred surface density of the molecular material continues the monotonic decrease from the inner regions. We conclude that although molecular clouds can form in the outer region of this galaxy, there is little mass associated with them. PMID- 15549099 TI - Electrical generation and absorption of phonons in carbon nanotubes. AB - The interplay between discrete vibrational and electronic degrees of freedom directly influences the chemical and physical properties of molecular systems. This coupling is typically studied through optical methods such as fluorescence, absorption and Raman spectroscopy. Molecular electronic devices provide new opportunities for exploring vibration-electronic interactions at the single molecule level. For example, electrons injected from a scanning tunnelling microscope tip into a metal can excite vibrational excitations of a molecule situated in the gap between tip and metal. Here we show how current directly injected into a freely suspended individual single-wall carbon nanotube can be used to excite, detect and control a specific vibrational mode of the molecule. Electrons tunnelling inelastically into the nanotube cause a non-equilibrium occupation of the radial breathing mode, leading to both stimulated emission and absorption of phonons by successive electron tunnelling events. We exploit this effect to measure a phonon lifetime of the order of 10 ns, corresponding to a quality factor of well over 10,000 for this nanomechanical oscillator. PMID- 15549100 TI - Random quasi-phase-matching in bulk polycrystalline isotropic nonlinear materials. AB - Three-wave mixing in nonlinear materials--the interaction of two light waves to produce a third--is a convenient way of generating new optical frequencies from common laser sources. However, the resulting optical conversion yield is generally poor, because the relative phases of the three interacting waves change continuously as they propagate through the material. This phenomenon, known as phase mismatch, is a consequence of optical dispersion (wave velocity is frequency dependent), and is responsible for the poor optical conversion potential of isotropic nonlinear materials. Here we show that exploiting the random motion of the relative phases in highly transparent polycrystalline materials can be an effective strategy for achieving efficient phase matching in isotropic materials. Distinctive features of this 'random quasi-phase-matching' approach are a linear dependence of the conversion yield with sample thickness (predicted in ref. 3), the absence of the need for either preferential materials orientation or specific polarization selection rules, and the existence of a wavelength-dependent resonant size for the polycrystalline grains. PMID- 15549101 TI - Metal wires for terahertz wave guiding. AB - Sources and systems for far-infrared or terahertz (1 THz = 10(12) Hz) radiation have received extensive attention in recent years, with applications in sensing, imaging and spectroscopy. Terahertz radiation bridges the gap between the microwave and optical regimes, and offers significant scientific and technological potential in many fields. However, waveguiding in this intermediate spectral region still remains a challenge. Neither conventional metal waveguides for microwave radiation, nor dielectric fibres for visible and near-infrared radiation can be used to guide terahertz waves over a long distance, owing to the high loss from the finite conductivity of metals or the high absorption coefficient of dielectric materials in this spectral range. Furthermore, the extensive use of broadband pulses in the terahertz regime imposes an additional constraint of low dispersion, which is necessary for compatibility with spectroscopic applications. Here we show how a simple waveguide, namely a bare metal wire, can be used to transport terahertz pulses with virtually no dispersion, low attenuation, and with remarkable structural simplicity. As an example of this new waveguiding structure, we demonstrate an endoscope for terahertz pulses. PMID- 15549102 TI - Constraints on the duration and freshwater release of Heinrich event 4 through isotope modelling. AB - Heinrich events--abrupt climate cooling events due to ice-sheet instability that occurred during the last glacial period--are recorded in sediment cores throughout the North Atlantic Ocean. Modelling studies have described likely physical mechanisms for these events, but the quantitative characteristics of Heinrich events are less well known. Here we use a climate model of intermediate complexity that explicitly calculates the distribution of oxygen isotopes in the oceans to simulate Heinrich event 4 at about 40,000 yr ago. We compare an ensemble of scenarios for this Heinrich event with oxygen isotope data measured in foraminiferal calcite of a comprehensive set of sediment cores. From this comparison, we obtain a duration of 250 +/- 150 yr and an ice release of 2 +/- 1 m sea-level equivalent for Heinrich event 4, significantly reducing the uncertainties in both values compared to earlier estimates of up to 2,000 yr and 15 m of sea-level equivalent ice release, respectively. Our results indicate that the consequences of Heinrich events may have been less severe than previously assumed, at least with respect to Greenland climate and sea level. PMID- 15549103 TI - Triassic marine reptiles gave birth to live young. AB - Sauropterygians form the largest and most diverse group of ancient marine reptiles that lived throughout nearly the entire Mesozoic era (from 250 to 65 million years ago). Although thousands of specimens of this group have been collected around the world since the description of the first plesiosaur in 1821 (ref. 3), no direct evidence has been found to determine whether any sauropterygians came on shore to lay eggs (oviparity) like sea turtles, or gave birth in the water to live young (viviparity) as ichthyosaurs and mosasauroids (marine lizards) did. Viviparity has been proposed for plesiosaur, pachypleurosaur and nothosaur sauropterygians, but until now no concrete evidence has been advanced. Here we report two gravid specimens of Keichousaurus hui Young from the Middle Triassic of China. These exquisitely preserved specimens not only provide the first unequivocal evidence of reproductive mode and sexual dimorphism in sauropterygians, but also indicate that viviparity could have been expedited by the evolution of a movable pelvis in pachypleurosaurs. By extension, this has implications for the reproductive pattern of other sauropterygians and Mesozoic marine reptiles that possessed a movable pelvis. PMID- 15549105 TI - An obligate brood parasite trapped in the intraspecific arms race of its hosts. AB - Reciprocal selection pressures often lead to close and adaptive matching of traits in coevolved species. A failure of one species to match the evolutionary trajectories of another is often attributed to evolutionary lags or to differing selection pressures across a geographic mosaic. Here we show that mismatches in adaptation of interacting species--an obligate brood parasitic duck and each of its two main hosts--are best explained by the evolutionary dynamics within the host species. Rejection of the brood parasite's eggs was common by both hosts, despite a lack of detectable cost of parasitism to the hosts. Egg rejection markedly reduced parasite fitness, but egg mimicry experiments revealed no phenotypic natural selection for more mimetic parasitic eggs. These paradoxical results were resolved by the discovery of intraspecific brood parasitism and conspecific egg rejection within the hosts themselves. The apparent arms race between species seems instead to be an incidental by-product of within-species conflict, with little recourse for evolutionary response by the parasite. PMID- 15549104 TI - The evolution of alternative parasitic life histories in large blue butterflies. AB - Large blue (Maculinea) butterflies are highly endangered throughout the Palaearctic region, and have been the focus of intense conservation research. In addition, their extraordinary parasitic lifestyles make them ideal for studies of life history evolution. Early instars consume flower buds of specific host plants, but later instars live in ant nests where they either devour the brood (predators), or are fed mouth-to-mouth by the adult ants (cuckoos). Here we present the phylogeny for the group, which shows that it is a monophyletic clade nested within Phengaris, a rare Oriental genus whose species have similar life histories. Cuckoo species are likely to have evolved from predatory ancestors. As early as five million years ago, two Maculinea clades diverged, leading to the different parasitic strategies seen in the genus today. Contrary to current belief, the two recognized cuckoo species show little genetic divergence and are probably a single ecologically differentiated species. On the other hand, some of the predatory morphospecies exhibit considerable genetic divergence and may contain cryptic species. These findings have important implications for conservation and reintroduction efforts. PMID- 15549106 TI - Spatial patterns in species distributions reveal biodiversity change. AB - Interpretation of global biodiversity change is hampered by a lack of information on the historical status of most species in most parts of the world. Here we show that declines and increases can be deduced from current species distributions alone, using spatial patterns of occupancy combined with distribution size. Declining species show sparse, fragmented distributions for their distribution size, reflecting the extinction process; expanding species show denser, more aggregated distributions, reflecting colonization. Past distribution size changes for British butterflies were deduced successfully from current distributions, and former distributions had some power to predict future change. What is more, the relationship between distribution pattern and change in British butterflies independently predicted distribution change for butterfly species in Flanders, Belgium, and distribution change in British rare plant species is similarly related to spatial distribution pattern. This link between current distribution patterns and processes of distribution change could be used to assess relative levels of threat facing different species, even for regions and taxa lacking detailed historical and ecological information. PMID- 15549107 TI - Identification of human brain tumour initiating cells. AB - The cancer stem cell (CSC) hypothesis suggests that neoplastic clones are maintained exclusively by a rare fraction of cells with stem cell properties. Although the existence of CSCs in human leukaemia is established, little evidence exists for CSCs in solid tumours, except for breast cancer. Recently, we prospectively isolated a CD133+ cell subpopulation from human brain tumours that exhibited stem cell properties in vitro. However, the true measures of CSCs are their capacity for self renewal and exact recapitulation of the original tumour. Here we report the development of a xenograft assay that identified human brain tumour initiating cells that initiate tumours in vivo. Only the CD133+ brain tumour fraction contains cells that are capable of tumour initiation in NOD-SCID (non-obese diabetic, severe combined immunodeficient) mouse brains. Injection of as few as 100 CD133+ cells produced a tumour that could be serially transplanted and was a phenocopy of the patient's original tumour, whereas injection of 10(5) CD133- cells engrafted but did not cause a tumour. Thus, the identification of brain tumour initiating cells provides insights into human brain tumour pathogenesis, giving strong support for the CSC hypothesis as the basis for many solid tumours, and establishes a previously unidentified cellular target for more effective cancer therapies. PMID- 15549108 TI - A FADD-dependent innate immune mechanism in mammalian cells. AB - Vertebrate innate immunity provides a first line of defence against pathogens such as viruses and bacteria. Viral infection activates a potent innate immune response, which can be triggered by double-stranded (ds)RNA produced during viral replication. Here, we report that mammalian cells lacking the death-domain containing protein FADD are defective in intracellular dsRNA-activated gene expression, including production of type I (alpha/beta) interferons, and are thus very susceptible to viral infection. The signalling pathway incorporating FADD is largely independent of Toll-like receptor 3 and the dsRNA-dependent kinase PKR, but seems to require receptor interacting protein 1 as well as Tank-binding kinase 1-mediated activation of the transcription factor IRF-3. The requirement for FADD in mammalian host defence is evocative of innate immune signalling in Drosophila, in which a FADD-dependent pathway responds to bacterial infection by activating the transcription of antimicrobial genes. These data therefore suggest the existence of a conserved pathogen recognition pathway in mammalian cells that is essential for the optimal induction of type I interferons and other genes important for host defence. PMID- 15549109 TI - Structure of a natural guanine-responsive riboswitch complexed with the metabolite hypoxanthine. AB - Riboswitches are genetic regulatory elements found in the 5' untranslated region of messenger RNA that act in the absence of protein cofactors. They are broadly distributed across bacteria and account for the regulation of more than 2% of all genes in Bacillus subtilis, underscoring their importance in the control of cellular metabolism. The 5' untranslated region of many mRNAs of genes involved in purine metabolism and transport contain a guanine-responsive riboswitch that directly binds guanine, hypoxanthine or xanthine to terminate transcription. Here we report the crystal structure at 1.95 A resolution of the purine-binding domain of the guanine riboswitch from the xpt-pbuX operon of B. subtilis bound to hypoxanthine, a prevalent metabolite in the bacterial purine salvage pathway. This structure reveals a complex RNA fold involving several phylogenetically conserved nucleotides that create a binding pocket that almost completely envelops the ligand. Hypoxanthine functions to stabilize this structure and to promote the formation of a downstream transcriptional terminator element, thereby providing a mechanism for directly repressing gene expression in response to an increase in intracellular concentrations of metabolite. PMID- 15549111 TI - Election returns. PMID- 15549112 TI - Putting pen to paper. PMID- 15549113 TI - Biosecurity with 'bio-sense'. PMID- 15549114 TI - Fifty years of interference. PMID- 15549115 TI - Vaccine availability in the US: problems and solutions. PMID- 15549116 TI - The death of a dogma? PMID- 15549117 TI - Lead and follow: the dance of the dendritic cell and T cell. PMID- 15549118 TI - AID: a very old motif newly recognized. PMID- 15549119 TI - Why are rabbits uniquely sensitive to myxoma virus? Cherchez l'interferon! PMID- 15549120 TI - Are dendritic cells afraid of commitment? PMID- 15549122 TI - Labeled antigens and antibodies: the evolution of magic markers and magic bullets. AB - The ability to label antigens and antibodies with simple chemicals and even with whole proteins fostered new approaches to basic studies of the immune system as well as new methods of immunodiagnosis and immunotherapy. This was especially true following the introduction of monoclonal antibodies, which enhanced the specificity of many of these applications. The uses to which these labeled immunoreagents were put were legion, and those who employed them might come from any field of biology or medicine. Many of these technical elaborations were critical to progress in immunology and in many other biomedical sciences. They illustrate also the often complex interplay between technology and theory. PMID- 15549123 TI - Plasmacytoid dendritic cells in immunity. AB - Human and mouse plasmacytoid dendritic cells have been shown to correspond to a specialized cell population that produces large amounts of type I interferons in response to viruses, the so-called natural interferon-producing cells. As a result, intensive investigation is now focused on the potential functions of plasmacytoid dendritic cells in both innate and adaptive immunity. Here we review recent progress on the characterization of plasmacytoid dendritic cell origin, development, migration and function in immunity and tolerance, as well as their effect on human diseases. PMID- 15549130 TI - Pharmacogenomics in admixed populations: the Brazilian pharmacogenetics/pharmacogenomics network--REFARGEN. PMID- 15549132 TI - Activity-dependent endocytic sorting of kainate receptors to recycling or degradation pathways. AB - Kainate receptors (KARs) play important roles in the modulation of neurotransmission and plasticity, but the mechanisms that regulate their surface expression and endocytic sorting remain largely unknown. Here, we show that in cultured hippocampal neurons the surface expression of GluR6-containing KARs is dynamically regulated. Furthermore, internalized KARs are sorted into recycling or degradative pathways depending on the endocytotic stimulus. Kainate activation causes a Ca2+- and PKA-independent but PKC-dependent internalization of KARs that are targeted to lysosomes for degradation. In contrast, NMDAR activation evokes a Ca2+-, PKA- and PKC-dependent endocytosis of KARs to early endosomes with subsequent reinsertion back into the plasma membrane. These results demonstrate that GluR6-containing KARs are subject to activity-dependent endocytic sorting, a process that provides a mechanism for both rapid and chronic changes in the number of functional receptors. PMID- 15549133 TI - Phosphorylation of XPB helicase regulates TFIIH nucleotide excision repair activity. AB - Nucleotide excision repair (NER) removes damage from DNA in a tightly regulated multiprotein process. The xeroderma pigmentosum group B (XPB) helicase subunit of TFIIH functions in NER and transcription. The serine 751 (S751) residue of XPB was found to be phosphorylated in vivo. This phosphorylation inhibits NER and the microinjection of a phosphomimicking XPB-S751E mutant is unable to correct the NER defect of XP-B cells. Conversely, XPB-S751 dephosphorylation or its substitution with alanine (S751A) restores NER both in vivo and in vitro. Surprisingly, phospho/dephosphorylation of S751 spares TFIIH-dependent transcription. Finally, the phosphorylation of XPB-S751 does not impair the TFIIH unwinding of the DNA around the lesion, but rather prevents the 5' incision triggered by the ERCC1-XPF endonuclease. These data support an additional role for XPB in promoting the incision of the damaged fragment and reveal a point of NER regulation on TFIIH without interference in its transcription activity. PMID- 15549134 TI - HAT cofactor Trrap regulates the mitotic checkpoint by modulation of Mad1 and Mad2 expression. AB - As a component of chromatin-modifying complexes with histone acetyltransferase (HAT) activity, TRRAP has been shown to be involved in various cellular processes including gene transcription and oncogenic transformation. Inactivation of Trrap, the murine ortholog of TRRAP, in mice revealed its function in development and cell cycle progression. However, the underlying mechanism is unknown. Here, we show that the loss of Trrap in mammalian cells leads to chromosome missegregation, mitotic exit failure and compromised mitotic checkpoint. These mitotic checkpoint defects are caused by defective Trrap-mediated transcription of the mitotic checkpoint proteins Mad1 and Mad2. The mode of regulation by Trrap involves acetylation of histones H4 and H3 at the gene promoter of these mitotic players. Trrap associated with the HAT Tip60 and PCAF at the Mad1 and Mad2 promoters in a cell cycle-dependent manner and Trrap depletion abolished recruitment of these HATs. Finally, ectopic expression of Mad1 and Mad2 fully restores the mitotic checkpoint in Trrap-deficient cells. These results demonstrate that Trrap controls the mitotic checkpoint integrity by specifically regulating Mad1 and Mad2 genes. PMID- 15549135 TI - The presenilin C-terminus is required for ER-retention, nicastrin-binding and gamma-secretase activity. AB - gamma-Secretase is an intramembrane cleaving protease involved in Alzheimer's disease. gamma-Secretase occurs as a high molecular weight complex composed of presenilin (PS1/2), nicastrin (NCT), anterior pharynx-defective phenotype 1 and PS enhancer 2. Little is known about the cellular mechanisms of gamma-secretase assembly. Here we demonstrate that the cytoplasmic tail of PS1 fulfills several functions required for complex formation, retention of unincorporated PS1 and gamma-secretase activity. The very C-terminus interacts with the transmembrane domain of NCT and may penetrate into the membrane. Deletion of the last amino acid is sufficient to completely block gamma-secretase assembly and release of PS1 from the endoplasmic reticulum (ER). This suggests that unincorporated PS1 is actively retained within the ER. We identified a hydrophobic stretch of amino acids within the cytoplasmic tail of PS1 distinct from the NCT-binding site, which is required to retain unincorporated PS1 within the ER. Deletion of the retention signal results in the release of PS1 from the ER and the assembly of a nonfunctional gamma-secretase complex, suggesting that at least a part of the retention motif may also be required for the function of PS1. PMID- 15549137 TI - The CDK regulates repair of double-strand breaks by homologous recombination during the cell cycle. AB - DNA double-strand breaks (DSBs) are dangerous lesions that can lead to genomic instability and cell death. Eukaryotic cells repair DSBs either by nonhomologous end-joining (NHEJ) or by homologous recombination. We investigated the ability of yeast cells (Saccharomyces cerevisiae) to repair a single, chromosomal DSB by recombination at different stages of the cell cycle. We show that cells arrested at the G1 phase of the cell cycle restrict homologous recombination, but are able to repair the DSB by NHEJ. Furthermore, we demonstrate that recombination ability does not require duplicated chromatids or passage through S phase, and is controlled at the resection step by Clb-CDK activity. PMID- 15549136 TI - Enterotoxigenic Escherichia coli vesicles target toxin delivery into mammalian cells. AB - Enterotoxigenic Escherichia coli (ETEC) is a prevalent cause of traveler's diarrhea and infant mortality in third-world countries. Heat-labile enterotoxin (LT) is secreted from ETEC via vesicles composed of outer membrane and periplasm. We investigated the role of ETEC vesicles in pathogenesis by analyzing vesicle association and entry into eukaryotic cells. Fluorescently labeled vesicles from LT-producing and LT-nonproducing strains were compared in their ability to bind adrenal and intestinal epithelial cells. ETEC-derived vesicles, but not control nonpathogen-derived vesicles, associated with cells in a time-, temperature-, and receptor-dependent manner. Vesicles were visualized on the cell surface at 4 degrees C and detected intracellularly at 37 degrees C. ETEC vesicle endocytosis depended on cholesterol-rich lipid rafts. Entering vesicles partially colocalized with caveolin, and the internalized vesicles accumulated in a nonacidified compartment. We conclude that ETEC vesicles serve as specifically targeted transport vehicles that mediate entry of active enterotoxin and other bacterial envelope components into host cells. These data demonstrate a role in virulence for ETEC vesicles. PMID- 15549138 TI - Peripheral relaxant activity of apomorphine and of a D1 selective receptor agonist on human corpus cavernosum strips. AB - Apomorphine is used in the erectile dysfunction therapy and its action has been ascribed to the stimulation of central dopamine receptor. At the present stage, very little is known about the peripheral action of apomorphine on human corpus cavernosum (HCC). We have investigated the peripheral action of apomorphine and the role of dopamine receptors in HCC. We here demonstrate that both D1 and D2 receptors were expressed in the HCC, D1 receptors were two-fold more abundant than D2 and that both receptors were mainly localized on the smooth muscle cell component. Apomorphine in vitro exerted an anti-alpha1 adrenergic activity in human cavernosal strips since it prevented contraction induced by phenylephrine (PE), but not by U46619 or endothelin. Apomorphine elicited endothelium independent and concentration-dependent relaxation of the strips contracted by PE, U46619 or endothelin. The EC50 values (microM) for apomorphine, in the presence and absence of endothelium, were 51.0+/-16 and 16.0+/-14, 120+/-19 and 150+/-18, 59.0+/-15 and 140+/-50 on PE-, U46619- or endothelin-induced contraction, respectively. Selective dopamine receptor agonist A-68930 (D1-like), but not quinpirole (D2-like), caused concentration-dependent relaxation of the cavernosal strips, which was partially prevented by endothelium removal or by treatment with an inhibitor of nitric oxide (NO) synthase. In conclusion, we show that (1) apomorphine has a peripheral relaxant direct effect as well as an antiadrenergic activity, (2) HCC possesses more D1-like (D1 and D5) than D2-like (D2, D3 and D4) receptors, (3) both D1- and D2-like receptors are mainly localized on smooth muscle cells and (4) the relaxant activity is most probably mediated by D1-like receptor partially through NO release from endothelium. PMID- 15549139 TI - Erectile dysfunction in chronic renal failure patients undergoing hemodialysis in Egypt. AB - In clinical practice, the attention given to sexual problems in patients with end stage renal disease is low. In order to evaluate the erectile function in chronic renal failure patients undergoing hemodialysis (HD) as a renal replacement therapy in upper Egypt, we used the abridged version of the International Index of Erectile Function (IIEF-5). In all, 75 HD patients were subjected to clinical and laboratory investigations. The controls were 948 healthy males representing the general Egyptian population. The prevalence of erectile dysfunction (ED) among the HD patients was 82.5% compared to 30% among controls. The prevalence of ED in HD group was significantly higher than in controls. The prevalence of ED in HD patients <50 y was 80% and it was 88% in those > or =50 y, while the prevalence of ED among controls was 28 and 69.8%, respectively. The prevalence of severe degree of ED was significantly higher in both groups compared to controls, while moderate degree of ED showed a statistical significance compared to controls in age groups <50 y and mild degree of ED showed a statistical significance compared to controls in age groups > or =50 y. [corrected] Age (r= 0.3368, P<0.01), serum urea (r=-0.5974, P<0.001), and creatinine level (r= 0.5804, P<0.001) have a significant negative correlation with the presence of ED among HD patients, while serum hemoglobin (r=0.3396, P<0.001) and years of HD age (r=0.3147, P<0.01) have a significant positive correlation with the presence of ED among the HD patients. In view of the observed high prevalence of ED among the HD patients, we believe that a complete health evaluation of male HD patients should include a discussion about erectile function in the standard clinical care program of patients with renal disease. PMID- 15549140 TI - Unresolved priapism secondary to tamsulosin. AB - Tamsulosin is the most potent adrenergic alpha-1 antagonist used for the treatment of benign prostatic hyperplasia. Priapism has been reported rarely in patients taking Prazosin, Doxazosin and Terazosin. We describe an otherwise healthy man with recurrent and then persistent unresolved priapism after the use of tamsulosin. Initial treatment consisted of aspiration and intracavernosal irrigation of iced saline and vasoconstrictive agent, but in vain. We then performed Winters procedure but that too failed and the priapism persisted. Health-care professionals should inform all patients taking such medications about rare but possible serious adverse effects. PMID- 15549141 TI - Diagnostic significance of transferrinuria and albumin-specific dipstick testing in primary care patients with elevated office blood pressure. AB - This study assesses the diagnostic accuracy of transferrinuria and an albumin specific dipstick assay for detection of renal target organ damage (microalbuminuria) in hypertensive patients in a general practice setting. A spot urine sample of 130 nondiabetic patients with elevated office blood pressure readings (>140 and/or 90 mmHg) was investigated by measuring albumin to creatinine ratio (ACR) and transferrin to creatinine ratio (TCR) and by using an albumin-specific dipstick test (Micral). ACR was considered as comparative gold standard. TCR was elevated (>0.19 mg/mmol) in 26 urine samples (20.0% of the test samples). ACR was raised in 29 samples (22.3% of the test samples). Elevated TCR had a sensitivity of 97% and specificity of 91% for detection of microalbuminuria. Positive predicting value for microalbuminuria was 65%; negative predicting value was 99%. Correlation between ACR and TCR was strong (r=0.96). Dipstick testing for albumin was positive in 23 urine samples (17.7% of the test samples), 27 (20.8%) tests were false positive and six (4.6%) false negative. When dipstick was positive, the sensitivity of detecting microalbuminuria was 79%, and specificity 73%. In conclusion, detection of urinary transferrin in nondiabetic patients with hypertension is strongly associated with urinary albumin excretion. However, assessment of TCR does not identify additional patients with microalbuminuria compared to measurement of ACR alone. The semiquantitative Micral test offers a simple and valuable method to screen hypertensive patients for microalbuminuria in a primary care setting. PMID- 15549142 TI - Angiotensin converting enzyme insertion/deletion polymorphism does not alter sepsis outcome in ventilated very low birth weight infants. AB - OBJECTIVE: This study compared the effect of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) polymorphisms on the incidence and outcome of sepsis in ventilated very low birth weight infants. STUDY DESIGN: Infectious complications were retrospectively determined in 295 (234 African-American, 58 Caucasian and three Hispanic) mechanically ventilated very low birth weight (VLBW) infants (<1500 g at birth) and compared ACE I/D genotype. RESULTS: The incidence of the D allele in the study population was 0.60. A total of 113 (38.3%) infants were homozygous DD, 128 (43.4%) were heterozygous ID and 54 (18.3%) were homozygous II. One or more episodes of late BSI developed in 28 (52%) of 54 infants with the II genotype, 66 (52%) of 128 infants with the ID genotype and 52 (46%) of 113 infants with the DD genotype (p=0.618). Neither the rates of non-CONS BSI (II: 24%, ID: 23%, DD: 22%; p=0.937) nor multiple bacteremic/fungemic episodes (II: 13%, ID: 16%, DD: 12%; p=0.641) were different between genotype groups. The ACE I/D polymorphism had no effect on sepsis-related mortality (II: 7%, ID: 5%, DD: 4%; p=0.692). Sepsis mortality for infants with late BSI was 14% in infants with the II genotype, 9% with the ID genotype and 10% with the DD genotype (p=0.764). CONCLUSIONS: The ACE I/D polymorphism does not have a significant effect on the incidence or outcome of sepsis in ventilated VLBW infants. PMID- 15549143 TI - Variability in the dose of intravenous vitamin E given to very low birth weight infants. AB - OBJECTIVE: Vitamin E is required by very low birth weight (VLBW) infants to prevent vitamin E deficiency. However, prolonged intravenous intakes of vitamin E >4 international units (IU)/kg/day often yield potentially toxic serum tocopherol levels. This study was designed to assess the frequency of potentially inadequate or excessive doses of intravenous vitamin E in parenteral nutrition for VLBW infants in the US. STUDY DESIGN: A questionnaire was sent to the 100 neonatal perinatal training program centers listed in the 2003 directory (American Academy of Pediatrics (AAP)). Using the information provided for each neonatal unit, we calculated the doses of vitamin E (IU/kg/day) that would have been given to infants with body weights of 0.5, 1.0 and 1.5 kg in that unit. The doses were then recoded as inadequate if less than recommended (2.8 IU/kg/day, maximum 7 IU/day) by the American Society for Clinical Nutrition and the AAP, and excessive if >4 IU/kg/day, and frequencies were compared using chi2 analysis. RESULTS: The 65 centers that responded were predominantly among those that offered the highest level of subspecialty neonatal intensive care (level IIID). The predicted dose of vitamin E had a median value of 2.8 IU/kg/day for all three weights, and was no significantly different among the three weights. VLBW infants with weights of 0.5, 1.0 and 1.5 kg were predicted to receive doses <2.8 IU/kg/day in 12, 12 and 19% of the neonatal intensive care units (NICUs), respectively, and doses >4 IU/kg/day in 40, 31 and 10% of the NICUs, respectively. Excessive doses were significantly associated with body weight, with more frequent excessive doses at lower weight (p=0.0008). CONCLUSION: This survey showed a lack of uniformity of dosing of intravenous vitamin E in VLBW infants. The smallest infants (<1.0 kg) were receiving excessive, potentially toxic doses of vitamin E in a significant number of NICUs in the US in 2003. PMID- 15549144 TI - Telomeres and telomerase in paediatric patients with T-cell acute lymphoblastic leukaemia (T-ALL). PMID- 15549145 TI - Acadesine induces apoptosis in B cells from mantle cell lymphoma and splenic marginal zone lymphoma. PMID- 15549146 TI - CTLA-4 overexpression in CD19+/CD5+ cells correlates with the level of cell cycle regulators and disease progression in B-CLL patients. PMID- 15549147 TI - WHO-defined chronic neutrophilic leukemia: a long-term analysis of 12 cases and a critical review of the literature. PMID- 15549148 TI - Tuberculosis incidence and mortality in aboriginal areas of Taiwan, 1997-2001. AB - BACKGROUND AND PURPOSE: People living in aboriginal areas of Taiwan are known to be at especially high risk of developing tuberculosis (TB). This study investigated the incidence and mortality of TB in aboriginal areas of Taiwan. METHODS: The TB death statistics and notification data from the National TB Register and Department of Health in 1997-2001 were analyzed. RESULTS: From 1997 to 2001, people living in aboriginal areas bore a disproportionate burden of TB disease in Taiwan, with mortality rates 5.5-6.5 times and incidence rates 3.6-5.2 times higher than those of people living in non-aboriginal areas. Among patients with TB living in aboriginal areas, 381 died, accounting for 5.1% of the 7480 TB deaths in Taiwan. A large proportion of the patients with TB who died were older than 65 years, in both aboriginal (40.2%) and non-aboriginal (78.5%) areas; however, the age of TB patients who died in non-aboriginal areas was significantly older (p < 0.001). TB patients living in the aboriginal areas accounted for 3.6% (2618/71,447) of the total number of reported TB patients in Taiwan, and 32.7% were in the age range from 24-45 years. In contrast, TB patients living in non-aboriginal areas were typically older, with only 20.1% in the 24-45 age range (p < 0.001) and 44.4% older than 65 years (p < 0.001). In terms of incidence and mortality rates, men predominated in both groups; however, this pattern was less prominent in aboriginal areas than in non-aboriginal areas. CONCLUSIONS: The mortality rates and incidence of TB in aboriginal areas are much higher than those in non-aboriginal areas in Taiwan. Concentration of resources and programs to control TB in aboriginal areas may represent the most effective use of resources for fighting TB. PMID- 15549149 TI - Clinical characteristics of patients undergoing surgical resection of benign liver tumors in Taiwan, an endemic area for hepatocellular carcinoma. AB - BACKGROUND AND PURPOSE: The management of benign liver tumors in Asian countries endemic for hepatocellular carcinoma (HCC) may be different from that in western countries with a lower prevalence of HCC. The aim of this study was to investigate the characteristics of surgically treated benign liver tumors in a liver disease treatment center in Taiwan, an area prevalent for HCC. METHODS: Between January 1991 and June 2001, 57 patients with benign liver tumors underwent liver resection. The demographic data, radiologic diagnosis, and postoperative pathologic diagnosis of these patients were reviewed and analyzed. RESULTS: Cavernous hemangioma (n = 15), focal nodular hyperplasia (n = 12), and macro-regenerative nodule (n = 9) were the most frequent pathologic diagnoses of the resected liver tumors. HCC was the most frequent preoperative radiologic diagnosis (27/57, 47%). Four of the 9 macro-regenerative nodules had associated focal dysplastic change. All of the patients with macro-regenerative nodule had a preoperative imaging diagnosis of HCC. Only 1 liver biopsy was performed among the 57 patients. CONCLUSIONS: HCC was the most frequent preoperative radiologic diagnosis for benign liver tumors undergoing liver resection at a tertiary referral center in Taiwan. Macro-regenerative nodule of liver was commonly misdiagnosed as HCC by radiology alone in this study. In order to avoid unnecessary surgery, aspiration cytology should be performed for all patients without elevated serum alpha-fetoprotein and typical radiologic features of HCC. Patients with macro-regenerative nodules without atypical hepatocytes can be followed safely without surgical intervention, but when a macro-regenerative nodule with dysplastic change is found, it should be treated either by surgical resection or local ablation. PMID- 15549150 TI - Acute acetaminophen intoxication in Taiwan: outcomes and risk factors. AB - BACKGROUND AND PURPOSE: There is a paucity of information about acetaminophen intoxication from Taiwan. This study investigated the outcome and risk factors for acetaminophen-induced hepatotoxicity and validated the Rumack- Matthew nomogram in Taiwanese patients with acute acetaminophen intoxication. METHODS: A total of 75 patients with acetaminophen intoxication admitted through the emergency department were included in this retrospective analysis. Patients with a serum acetaminophen concentration above the possible risk line on the nomogram were treated with oral N-acetylcysteine. The primary outcome measure was the development of major hepatotoxicity, which was defined as a serum aminotransferase concentration greater than 1000 IU/L. Patient outcomes in the possible risk group and probable risk group were plotted on the modified Rumack Matthew nomogram for validation. The risk factors for acetaminophen-induced hepatotoxicity were identified by multiple logistic regression analysis. RESULTS: No hepatotoxicity developed in patients with an initial acetaminophen concentration below the possible risk line on the nomogram. One out of 8 patients in the possible risk group developed major hepatotoxicity; 8 out of 22 patients in the probable risk group developed major hepatotoxicity, representing an incidence of 12.5% and 36.4%, respectively. Patients in the major hepatotoxicity group were older (32.5 vs 24.2 years, p = 0.019), and had a longer time to presentation (28.1 vs 6.7 hours, p < 0.01) than those in the non/minor hepatotoxicity group. Multiple logistic regression revealed that age and time to presentation were independent risk factors for hepatotoxicity (p = 0.033 and p = 0.002, respectively). CONCLUSIONS: The results of outcome analysis confirm that the modified Rumack-Matthew nomogram has a high sensitivity for identifying Taiwanese patients at risk for acetaminophen-induced hepatoxicity. Patient age and time to presentation were independent risk factors for acetaminophen-induced hepatotoxicity. PMID- 15549151 TI - Biomechanical and clinical effects of adding interfragmentary wiring to locked nailing for humeral nonunions. AB - BACKGROUND AND PURPOSE: The use of locked nailing for the treatment of humeral nonunions is threatened by the potential of initial instability which may result in residual fracture gap and motion and is detrimental to fracture healing. This study investigated the biomechanical and clinical effects of adding interfragmentary wiring to reduce the initial instability in the treatment of humeral nonunion. METHODS: The biomechanical study compared the mechanical properties of 3 fixation methods: locked nail alone, with staple, and with interfragmentary wiring. Composite humeri were transected to simulate nonunion and were retrograde nailed. Nail-bone constructs were examined by non-destructive bending tests and destructive torsional tests. The initial instability, fixation stiffness and strength were measured and compared. The clinical study compared 21 nonunions treated by locked nailing alone and 30 nonunions treated by locked nailing with interfragmentary wiring. The indication for wiring was persistent nonunion gap or motion during operation. The union rate, time to union, functional recovery of the adjacent joints and the time for functional recovery were compared between the treatment groups. RESULTS: Biomechanically, in both bending and torsional tests, locked nailing with a staple or wiring resulted in significantly less initial instability than locked nailing alone. Test results for bending stiffness, torsional stiffness, or torsional strength did not differ significantly among the 3 fixation methods. In the clinical study, interfragmentary wiring had the advantage of providing tight nonunion compression, which was beneficial for nonunion healing and promoted early postoperative exercise. Patients treated with locked nailing with interfragmentary wiring had significantly shorter time to union and faster shoulder functional recovery (p < 0.05). CONCLUSIONS: Adding interfragmentary wiring was an effective and safe procedure in humeral nonunions treated with locked nailing. PMID- 15549152 TI - Clinical manifestations, disease course, and complications of adult-onset Still's disease in Taiwan. AB - BACKGROUND AND PURPOSE: Data on the disease course and ultimate outcome of adult onset Still's disease (AOSD) are limited. We analyzed the clinical manifestations, disease course, and complications of patients with AOSD in Taiwan. METHODS: A retrospective cohort design with prospective follow-up was used. Eighty two patients with AOSD diagnosed between 1983 and 2003 were evaluated. Their clinical features and laboratory findings at presentation, disease course, and complications were analyzed. RESULTS: Fifty nine patients (72%) were female and 55 (67.1%) were aged between 16-35 years at onset. The most common clinical manifestations were fever (100%), articular symptoms (100%), evanescent rash (87%), and sore throat (84%). Dermatographism was noted in 59% of patients. Elevation of erythrocyte sedimentation rate and C-reactive protein, which were significantly correlated with disease activity score (both p < 0.01) occurred in more than 90% of AOSD patients. Elevation of serum ferritin, which was significantly correlated with disease activity score and hepatic enzyme levels, was present in 91% of patients. Polycyclic systemic course was the most common (45%), followed by monocyclic systemic course (34%); only 20% of patients progressed to chronic arthropathy. CONCLUSIONS: The multisystemic involvement and various patterns of disease course in this series illustrate the heterogenic nature of AOSD. Serum ferritin levels can be used as a marker for monitoring disease activity in AOSD. PMID- 15549153 TI - Partial duplication of 3q and distal deletion of 10q inherited from a maternal balanced translocation. AB - Partial trisomy 3q syndrome is often the result of an unbalanced translocation or inversion. The duplicated segments are mostly from 3q25 to 3qter. We describe a karyotype of 46,XY,der(10)t(3;10)(q25.3;q26.1) in a 1-day-old male infant who presented with multiple congenital anomalies including synophrys, a long philtrum, thin lips with down-turned angles of the mouth, micrognathia, a high arched and cleft palate, clenched hands, genital hypoplasia, cryptorchidism, a large ventricular septal defect, a subependymal cyst, and corpus callosum hypoplasia. The patient had cardiopulmonary distress resulting from multiple congenital anomalies. He died of heart failure at the age of 18 days. The chromosome aberration resulted from a maternal balanced translocation. The dup(3q) syndrome superficially resembles but can be distinguished from Brachmann de Lange syndrome. Craniofacial features, cleft palate and urinary tract anomaly are more frequent in dup(3q) syndrome. Oligodactyly and phocomelia are more characteristic of Brachmann-de Lange syndrome. PMID- 15549154 TI - Acute urethritis and arthritis-dermatitis syndrome due to Neisseria meningitidis. AB - Acute urethritis and arthritis-dermatitis syndrome after sexual contact are often assumed to be caused by Neisseria gonorrhoeae. We report a case of arthritis dermatitis syndrome in a 32-year-old man who presented with generalized maculopapular and petechial skin lesions and polyarthritis. Acute urethritis developed 1 week after oro-genital sexual contact with a sex worker about 3 weeks before admission. No pathogen was found on smear of urethral discharge and skin lesions, but Gram-negative diplococci were noted in joint fluid, and blood culture yielded N. meningitidis. His condition improved gradually after repeated arthrocentesis and antibiotic therapy with ceftriaxone followed by ciprofloxacin. Oro-genital contact is a transmission route for N. meningitidis infection manifesting as acute urethritis and arthritis-dermatitis syndrome. PMID- 15549155 TI - Congenital adrenal hyperplasia with 11 beta-hydroxylase deficiency. AB - A rare form of congenital adrenal hyperplasia (CAH), 11 beta-hydroxylase deficiency, may be misdiagnosed as 21-hydroxylase deficiency, the most common form of CAH, because of similar clinical presentations at times and elevated level of 17-hydroxyprogesterone in both conditions. We report a case of 11 beta hydroxylase deficiency that was originally misdiagnosed as 21-hydroxylase deficiency. Hypertension and hypokalemia complicated with seizures and arrhythmia developed in this 9-year-old girl after abrupt withdrawal of oral dexamethasone but maintenance of fludrocortisone. Suspicion of 11 beta-hydroxylase deficiency led to DNA mutation analysis, which revealed a novel point mutation (CTG 461 CCG) in the CYP11B1 gene converting leucine to proline. Her condition stabilized rapidly after withdrawal of fludrocortisone and administration of hydrocortisone. Regular measurement of blood pressure should be performed in all patients with CAH and test of serum 11-deoxycortisol or deoxycorticosterone level should be performed in those patients with elevated blood pressure to avoid misdiagnosis of 11 beta-hydroxylase deficiency. PMID- 15549156 TI - Dengue hemorrhagic fever complicated with acute pancreatitis and seizure. AB - Dengue fever is an acute febrile viral disease, which frequently presents with high fever, headache, bone pain and skin rash. Acute pancreatitis and seizure are rare manifestations of dengue virus infection. A 66-year-old woman with diabetes mellitus presented with epigastralgia, nausea, vomiting, diarrhea and fever. Acute pancreatitis, abnormal liver function and thrombocytopenia were diagnosed at a local hospital. After persistent fever, thrombocytopenia and seizure developed she was transferred to our medical center. Dengue virus infection was confirmed by serology study and dengue hemorrhagic fever grade II was diagnosed. No further neurological symptoms occurred and pancreatitis improved gradually after supportive care. She recovered and had no sequelae at 1 year follow-up. Acute pancreatitis and seizure may be manifestations of dengue virus infection, especially in patients with delayed diagnosis, prolonged fever and thrombocytopenia. PMID- 15549157 TI - Intra-retroperitoneal duplication cyst. AB - Duplication cyst occupying the retro- and intra-peritoneal space is a rare condition. We describe a case of duplication cyst in a 13-year-old girl who presented with abdominal pain, vomiting, and a lower abdominal mass. Plain abdominal X-ray films revealed local ileus over the lower abdomen. Ultrasonography revealed 2 double-layered cystic masses over the lower abdomen with a suspicious communicating tract. Mild hydropelvis of the right kidney was also noted. Abdominal computed tomography revealed 2 cystic lesions. One was located at the pelvic cavity just above the urinary bladder and the other was in the left retroperitoneal space. Laparotomy revealed a dumbbell-shaped intra retroperitoneal duplication cyst with a small communicating tract. The cyst was excised without disturbing bowel continuity and the vascular supply. The patient was doing well at 1-year follow-up. PMID- 15549158 TI - Neodymium:YAG laser incision of congenital obstructive posterior urethral membrane in boys with urinary incontinence and low uroflow. AB - BACKGROUND AND PURPOSE: Congenital obstructive posterior urethral membrane (COPUM) is associated with sub-clinical lower urinary tract symptoms in boys. Urethral stricture following cystoscopic fulguration of the membrane and a relatively small urethral caliber make the treatment of this condition difficult. METHODS: Fifteen boys (mean age, 10.0 years) with COPUM were treated with neodymium:YAG (Nd:YAG) contact laser incision of the membrane through a cystourethroscope (n = 7) or a ureteroscope (n = 8). RESULTS: Presenting symptoms were bedtime incontinence in 15, frequency/urgency in 10, daytime incontinence in 7, lazy bladder in 3 and urinary tract infection in 2. All patients had an obstructive urinary flow pattern with a maximal flow rate (Q(max)) < 15 mL/s. The mean follow-up period was 10.5 months. After the surgery, the number of wet nights per week reduced > 90%, 50-90% and < 50% in 9 (60%), 5 (33.3%), and 1 (6.7%), respectively. Mean (Q(max)) increased from 11.3 to 14.9 mL/s (p = 0.003). CONCLUSION: Nd:YAG laser incision of COPUM with the aid of a ureteroscope or a cystourethroscope is a safe and effective method for improving urinary flow and bedtime incontinence. PMID- 15549159 TI - Dynamic evaluation of 18F-FDG uptake by microPET and whole-body autoradiography in a fibrosarcoma-bearing mouse model. AB - BACKGROUND AND PURPOSE: Fluorine-18-2-deoxy-D-glucose (18F-FDG) has been used in the clinic as a diagnostic radiotracer for monitoring many kinds of tumors, but its value for monitoring fibrosarcoma is not well established. METHODS: In this study, the uptake of 18F-FDG in a fibrosarcoma-bearing mouse model was evaluated using the high resolution positron emission tomography (PET) system microPET. Tumor cells were implanted in 3 FVB/N mice, and static microPET scanning was performed on day 1, 7, 12 and 15 after implantation. A dynamic microPET image was scanned on day 12 to determine the 18F-FDG uptake in 3 other tumor-bearing mice. Time-activity curves were plotted by drawing regions of interest in the tumor, liver, kidneys and muscles. The mice were sacrificed after dynamic microPET imaging and whole-body autoradiography (WBAR) was performed. For biodistribution study, 9 tumor-bearing mice, 3 per experimental group, were studied at 3 time points and the results were compared with the static microPET images. RESULTS: MicroPET images suggested that 18F-FDG could be used to monitor the growth of tumors 7 days after implantation. Dynamic scans of 18F-FDG uptake reached a plateau in the tumor after 20 minutes on day 12 after implantation. Both microPET and WBAR revealed evidence of tumor necrosis. The results of biodistribution and WBAR agreed with those from microPET images. CONCLUSION: MicroPET was useful for monitoring the growth of fibrosarcoma and determination of the maximal uptake time point of 18F-FDG in tumors in this tumor-bearing mouse model. PMID- 15549160 TI - [Biological vascularized matrix (BioVaM): a new method for solving the perfusion problems in tissue engineering]. AB - A new technique is presented to harvest an acellular matrix from a porcine small bowel segment preserving the mesenteric arterial and venous pedicles. Reseeding of this biological vascularized matrix (BioVaM) with functional cells, i.e. smooth muscle and urothelial cells isolated from the urinary tract, and resurfacing of its vascular structures with endothelial precursor cells results in a vascularized tissue engineered graft for reconstruction and augmentation of the urinary bladder. First promising short term implantation experiments using a porcine model for the evaluation of early graft perfusion after vascular anastomosis are presented. PMID- 15549161 TI - [Stem cell therapy for urinary incontinence]. AB - Experimental and clinical studies investigated whether urinary incontinence can be effectively treated with transurethral ultrasound-guided injections of autologous myoblasts and fibroblasts.This new therapy was performed in eight female pigs. It could be shown that the injected cells survived well and that new muscle tissue was formed. Next, 42 patients (29 women, 13 men) suffering from urinary stress incontinence were treated. The fibroblasts were mixed with a small amount of collagen as carrier material and injected into the urethral submucosa to treat atrophies of the mucosa. The myoblasts were directly injected into the rhabdosphincter to reconstruct the muscle and to heal morphological and functional defects. In 35 patients urinary incontinence could be completely cured. In seven patients who had undergone multiple surgical procedures and radiotherapy urinary incontinence improved. No side effects or complications were encountered postoperatively. The experimental as well as the clinical data clearly demonstrate that urinary incontinence can be treated effectively with autologous stem cells. The present data support the conclusion that this new therapeutic concept may represent a very promising treatment modality in the future. PMID- 15549167 TI - What's new in the renin-angiotensin system? PMID- 15549162 TI - [Tissue engineering of erectile nerves]. AB - Dissection of the cavernous nerves eliminates spontaneous erections and may lead to irreversible erectile dysfunction due to degeneration of cavernous tissue. Novel procedures to reconstruct penile innervation include cavernous nerve interposition grafting and neurotrophic treatments to revitalize penile neural input, evaluated thus far in various preclinical models of cavernous nerve injury. Schwann cells crucially contribute to successful axonal regeneration by mechanical and paracrine mechanisms in the injured nerve, and Schwann cells seeded into guidance channels have been successfully employed to support regeneration in animal models of cavernous nerve injury. Gene therapy, tissue engineering, and reconstructive techniques have been combined to deliver neurotrophic factors and recover erectile function. PMID- 15549168 TI - Structure of angiotensin I-converting enzyme. AB - Angiotensin-converting enzyme (ACE) is a zinc- and chloride-dependent metallopeptidase that plays a vital role in the metabolism of biologically active peptides. Until recently, much of the inhibitor design and mechanism of action of this ubiquitous enzyme was based on the structures of carboxypeptidase A and thermolysin. When compared to the recently solved structures of the testis isoform of ACE (tACE) and its Drosophila homologue (AnCE), carboxypeptidase A showed little structural homology outside of the active site, while thermolysin revealed significant but less marked overall similarity. The ellipsoid-shaped structure of tACE, which has a preponderance of alpha-helices, is characterised by a core channel that has a constriction approximately 10 A from its opening where the zinc-binding active site is located. Comparison of the native protein with the inhibitor-bound form (lisinopril-tACE) does not reveal any striking differences in the conformation of the inhibitor binding site, disfavouring an open and closed configuration. However, the inhibitor complex does provide insights into the network of hydrogen-bonding and ionic interactions in the active site as well as the mechanism of ACE substrate hydrolysis. The three dimensional structure of ACE now paves the way for the rational design of a new generation of domain-selective ACE inhibitors. PMID- 15549169 TI - When 6 is 9: 'uncoupled' AT1 receptors turn signalling on its head. AB - The type 1 angiotensin receptor (AT(1)) activates an array of intracellular signalling pathways that control cell and tissue responses to the peptide hormone angiotensin II (AngII). The capacity of AT(1) receptors to initiate and maintain such signals has typically been explained on the basis of conventional heterotrimeric guanine nucleotide binding protein (G protein) activation, specifically G(q/11). Accumulating evidence from studies utilising a variety of AT(1) receptor mutants and AngII analogues indicates that some important downstream effects of AT(1) receptors are independent of classical G protein coupling. Importantly, AT(1) receptor-mediated endocytosis, tyrosine phosphorylation signalling and mitogen-activated protein kinase activation as well as transactivation of the epidermal growth factor receptor can occur in G(q/11)-uncoupled receptor mutants. These observations point to a functional partitioning of AT(1) receptor signals that permits separation of short-term AngII actions (e.g., vasoconstriction) from more extended events, such as pathological cell growth in heart and blood vessels, and may open up new avenues for selective antagonism. PMID- 15549170 TI - Hijacking epidermal growth factor receptors by angiotensin II: new possibilities for understanding and treating cardiac hypertrophy. AB - Activation of the type 1 angiotensin II receptor (AT(1)R) is associated with the aetiology of left ventricular hypertrophy, although the exact intracellular signalling mechanism(s) remain unclear. Transactivation of the epidermal growth factor receptor (EGFR) has emerged as a central mechanism by which the G protein coupled AT(1)R, which lacks intrinsic tyrosine kinase activity, can stimulate the mitogen-activated protein kinase signalling pathways thought to mediate cardiac hypertrophy. Current studies support a model whereby AT(1)R-dependent transactivation of EGFRs on cardiomyocytes involves stimulation of membrane-bound metalloproteases, which in turn cleave EGFR ligands such as heparin-binding EGF from a plasma membrane-associated precursor. Numerous aspects of the 'triple membrane-passing signalling' paradigm of AT(1)R-induced EGFR transactivation remain to be characterised, including the identity of the specific metalloproteases involved, the intracellular mechanism for their activation and the exact EGFR subtypes required. Here we examine how 'hijacking' of the EGFR might explain the ability of the AT(1)R to elicit the temporally and qualitatively diverse responses characteristic of the hypertrophic phenotype, and discuss the ramifications of delineating these pathways for the development of new therapeutic strategies to combat cardiac hypertrophy. PMID- 15549171 TI - Angiotensin-converting enzyme-2: a molecular and cellular perspective. AB - Angiotensin-converting enzyme-2 (ACE2) is the first human homologue of ACE to be described. ACE2 is a type I integral membrane protein which functions as a carboxypeptidase, cleaving a single hydrophobic/basic residue from the C-terminus of its substrates. ACE2 efficiently hydrolyses the potent vasoconstrictor angiotensin II to angiotensin (1-7). It is a consequence of this action that ACE2 participates in the renin-angiotensin system. However, ACE2 also hydrolyses dynorphin A (1-13), apelin-13 and des-Arg(9) bradykinin. The role of ACE2 in these peptide systems has yet to be revealed. A physiological role for ACE2 has been implicated in hypertension, cardiac function, heart function and diabetes, and as a receptor of the severe acute respiratory syndrome coronavirus. This paper reviews the biochemistry of ACE2 and discusses key findings such as the elucidation of crystal structures for ACE2 and testicular ACE and the development of ACE2 inhibitors that have now provided a basis for future research on this enzyme. PMID- 15549172 TI - Physiological roles of angiotensin-converting enzyme 2. AB - Angiotensin-converting enzyme 2 (ACE2) is a recently discovered homologue of the key enzyme of the renin-angiotensin system, the angiotensin-converting enzyme. The ACE2 enzyme is mainly expressed in cardiac blood vessels and tubular epithelia of the kidneys. Together with ACE2's unique metallocarboxypeptidase activity, the restricted tissue distribution suggests a distinctive physiological function in blood pressure, blood flow and fluid regulation. The ace2 gene was mapped to quantitative trait loci affecting susceptibility to hypertension in rats. Furthermore, ACE2 appears to be a negative regulator of ACE in the heart. ACE2 messenger RNA and protein levels are substantially regulated in the kidney of diabetic and pregnant rats. The mechanism of ACE2 function and its physiologic significance are not yet fully understood; however, as ACE2 differs in its specificity and physiological role from ACE, this opens a new potential venue for drug discovery aimed at cardiovascular disease, hypertension and diabetic complications. PMID- 15549173 TI - Novel angiotensin peptides. AB - Virtually all existing evidence on the function of angiotensin II (Ang II) in the regulation of tissue homeostasis and blood pressure regulation bears on the more restricted question of what other mechanisms or systems may amplify or inhibit the actions of this important peptide. Whereas there is evidence that Ang II may potentiate the effects of catecholamines, various cytokines and also growth factors, the repertoire of substances which may inhibit the actions of Ang II is more limited and has been restricted primarily to prostacyclin, bradykinin and nitric oxide. Advances in receptor pharmacology and introduction of selective antagonists to two of the receptor subtypes at which Ang II binds permitted a more critical examination of the functions of the renin angiotensin system in physiological and pathophysiological conditions, as well as uncovering the previously unsuspected possibility that within the biochemical pathways leading to the formation of the peptide the renin angiotensin system could process either its immediate precursor (angiotensin I) or the actual Ang II peptide into an alternative form, angiotensin-(1-7) [Ang-(1-7)], the function of which was to antagonize the effects of Ang II. We review here the biological actions of Ang-(1 7) and discuss how this discovery may change altogether the perception of how the renin angiotensin system functions in the regulation of tissue perfusion pressure and the regulation of salt and water metabolism. PMID- 15549174 TI - The angiotensin IV/AT4 receptor. AB - The angiotensin AT(4) receptor was originally defined as the specific, high affinity binding site for the hexapeptide angiotensin IV (Ang IV). Subsequently, the peptide LVV-hemorphin 7 was also demonstrated to be a bioactive ligand of the AT(4) receptor. Central administration of Ang IV, its analogues or LVV-hemorphin 7 markedly enhance learning and memory in normal rodents and reverse memory deficits observed in animal models of amnesia. The AT(4) receptor has a broad distribution and is found in a range of tissues, including the adrenal gland, kidney, lung and heart. In the kidney Ang IV increases renal cortical blood flow and decreases Na(+) transport in isolated renal proximal tubules. The AT(4) receptor has recently been identified as the transmembrane enzyme, insulin regulated membrane aminopeptidase (IRAP). IRAP is a type II integral membrane spanning protein belonging to the M1 family of aminopeptidases and is predominantly found in GLUT4 vesicles in insulin-responsive cells. Three hypotheses for the memory-potentiating effects of the AT(4) receptor/IRAP ligands, Ang IV and LVV-hemorphin 7, are proposed: (i) acting as potent inhibitors of IRAP, they may prolong the action of endogenous promnestic peptides; (ii) they may modulate glucose uptake by modulating trafficking of GLUT4; (iii) IRAP may act as a receptor, transducing the signal initiated by ligand binding to its C-terminal domain to the intracellular domain that interacts with several cytoplasmic proteins. PMID- 15549175 TI - Angiotensin-converting enzyme 2: a functional receptor for SARS coronavirus. AB - Cellular entry of enveloped viruses is often dependent on attachment proteins expressed on the host cell surface. Viral envelope proteins bind these receptors, and, in an incompletely understood process, facilitate fusion of the cellular and viral membranes so as to introduce the viral core into the cytoplasm. Only a small fraction of viral receptors have been identified so far. Recently, a novel coronavirus was identified as the etiological agent of severe acute respiratory syndrome (SARS). The fusion protein gene of SARS coronavirus (SARS-CoV) was cloned and characterized, and shortly thereafter, angiotensin-converting enzyme 2 (ACE2) was shown to be its functional receptor. Identification of ACE2 as a receptor for SARS-CoV will likely contribute to the development of antivirals and vaccines. It may also contribute to the development of additional animal models for studying SARS pathogenesis, and could help identify the animal reservoir of SARS-CoV. PMID- 15549176 TI - The search for a unified theory of coagulation and inflammation. PMID- 15549177 TI - Alexander disease: past and present. PMID- 15549178 TI - Identification and characterization of a highly thermostable bacteriophage lysozyme. AB - Pseudomonas aeruginosa bacteriophage phiKMV is a T7-like lytic phage. Liquid chromatography-mass spectrometry of the structural proteins revealed gene product 36 (gp36) as part of the phiKMV phage particle. The presence of a lysozyme domain in the C terminal of this protein (gp36C) was verified by turbidimetric assays on chloroform-treated P. aeruginosa PAO1 and Escherichia coli WK6 cells. The molecular mass (20,884 Da) and pI (6.4) of recombinant gp36C were determined, as were the optimal enzymatic conditions (pH 6.0 in 16.7 mM phosphate buffer) and activity (4800 U/mg). Recombinant gp36C is a highly thermostable lysozyme, retaining 26% of its activity after 2 h at 100 degrees C and 21% after autoclaving. This thermostability could prove an interesting characteristic for food conservation technology. PMID- 15549179 TI - Identification of chicken lysozyme g2 and its expression in the intestine. AB - Lysozyme is an important component of the innate immune system, protecting the gastrointestinal tract from infection. The aim of the present study was to determine if lysozyme is expressed in the chicken ( Gallus gallus) intestine and to characterise the molecular forms expressed. Immunohistochemical staining localised lysozyme to epithelial cells of the villous epithelium along the length of the small intestine. There was no evidence for lysozyme expression in crypt epithelium and no evidence for Paneth cells. Immunoblots of chicken intestinal protein revealed three proteins: a 14-kDa band consistent with lysozyme c, and two additional bands of approximately 21 and 23 kDa, the latter consistent with lysozyme g. RT-PCR analyses confirmed that lysozyme c mRNA is expressed in 4-day, but not older chicken intestine and lysozyme g in 4- to 35-day chicken intestine. A novel chicken lysozyme g2 gene was identified by in silico analyses and mRNA for this lysozyme g2 was identified in the intestine from chickens of all ages. Chicken lysozyme g2 shows similarity with fish lysozyme g, including the absence of a signal peptide and cysteines involved in disulphide bond formation of the mammalian and bird lysozyme g proteins. Analyses using SecretomeP predict that chicken lysozyme g2 may be secreted by the non-classical secretory pathway. We conclude that lysozyme is expressed in the chicken small intestine by villous enterocytes. Lysozyme c, lysozyme g and g2 may fulfil complimentary roles in protecting the intestine. PMID- 15549180 TI - Administration of the antitumor drug mitoguazone protects normal thymocytes against spontaneous and etoposide-induced apoptosis. AB - The suggestion has been made that polyamines may be involved in the control of cell death, since exceedingly high or low levels induce apoptosis in different cell systems. For a deeper insight into the relationship between apoptosis and polyamine metabolism, we investigated in vitro the effect on rat thymocytes of mitoguazone (MGBG, which inhibits S-adenosylmethionine decarboxylase, i.e. a key enzyme in the polyamine biosynthetic pathway). Thymocytes were selected as an especially suitable model system, since they undergo spontaneous apoptosis in vivo and can be easily induced to apoptose in vitro by etoposide, used here as an apoptogenic agent. MGBG protected thymocytes from both spontaneous and drug induced apoptosis, and this protective effect was associated with a decrease in polyamine oxidase activity and total polyamine levels. PMID- 15549181 TI - Isolation of human epidermal stem cells by adherence and the reconstruction of skin equivalents. AB - The isolation of human epidermal stem cells is critical for their clinical applications. In the present study, we isolated three populations of epidermal keratinocytes according to their ability to adhere to collagen type IV: i.e., rapidly adhering (RA), slowly adhering (SA), and non-adhering (NA) cells. The aim of this study was to characterize RA cells and to investigate the possibility of using these cells for epidermis reconstruction. To identify RA cells, flow cytometric analysis was performed using anti-alpha(6) integrin and anti-CD71 antibodies. RA cells express high levels of alpha(6) integrin and low levels of CD71, which are considered as markers of an epidermal stem cell nature. Furthermore, electron microscopy showed that RA cells are small and have a high nuclear to cytoplasmic ratio, whereas SA and NA cells have well-developed cellular organelles and abundant tonofilaments. Western blot analysis showed that RA cells are slow cycling and express p63, a putative epidermal stem cell marker, whereas SA and NA cells express c-Myc, which is known to regulate stem cell fate. To compare epidermal regenerative abilities, skin equivalents (SEs) were made using RA, SA, and NA cells. The epidermis constructed from RA cells was well formed compared to those formed from SA or NA cells. In addition, only SEs with RA cells expressed alpha(6) integrin and beta(1) integrin at the basal layer. These results indicate that RA cells represent epidermal stem cells and are predominately comprised of stem cells. Therefore, the isolation of RA cells using a simple technique offers a potential route to their clinical application, because they are easily isolated and provide a high yield of epidermal stem cells. PMID- 15549182 TI - C-peptide stimulates Na+, K+-ATPase via activation of ERK1/2 MAP kinases in human renal tubular cells. AB - Proinsulin-connecting peptide (C-peptide) exerts physiological effects partially via stimulation of Na(+), K(+)-ATPase. We determined the molecular mechanism by which C-peptide stimulates Na(+), K(+)-ATPase in primary human renal tubular cells (HRTCs). Incubation of the cells with 5 nM human C-peptide at 37 degrees C for 10 min stimulated (86)Rb(+) uptake by 40% (p<0.01). The carboxy-terminal pentapeptide was found to elicit 57% of the activity of the intact molecule. In parallel with ouabain-sensitive (86)Rb(+) uptake, C-peptide increased alpha subunit phosphorylation and basolateral membrane (BLM) abundance of the Na(+), K(+)-ATPase alpha(1) and beta(1) subunits. The increase in BLM abundance of the Na(+), K(+)-ATPase alpha(1) and beta(1) subunits was accompanied by depletion of alpha(1) and beta(1) subunits from the endosomal compartments. C-peptide action on Na(+), K(+)-ATPase was ERK1/2-dependent in HRTCs. C-peptide-stimulated Na(+), K(+)-ATPase activation, phosphorylation of alpha(1)-subunit and translocation of alpha(1) and beta(1) subunits to the BLM were abolished by a MEK1/2 inhibitor (20 muM PD98059). C-peptide stimulation of (86)Rb(+) uptake was also abolished by preincubation of HRTCs with an inhibitor of PKC (1 muM GF109203X). C-peptide stimulated phosphorylation of human Na(+), K(+)-ATPase alpha subunit on Thr-Pro amino acid motifs, which form specific ERK substrates. In conclusion, C-peptide stimulates sodium pump activity via ERK1/2-induced phosphorylation of Thr residues on the alpha subunit of Na(+), K(+)-ATPase. PMID- 15549184 TI - Management of anemia in patients undergoing curative radiotherapy. Erythropoietin, transfusions, or better nothing? AB - BACKGROUND AND RESULTS: Anemia is a well-known risk factor for decreased local control and survival in patients undergoing curative radiotherapy. There is clear evidence from recent clinical investigations that anemia is an independent risk factor and hemoglobin (Hb) levels during radiotherapy are important (and not pretreatment Hb levels). The most likely explanation for the prognostic impact is the association with tumor hypoxia. An "optimal" Hb range with regard to tumor oxygenation seems to exist, and Hb levels < 11 g/dl and > approximately 15 g/dl impair tumor oxygenation but have (over a broader range) no significant impact on normal tissue oxygenation. There is some evidence from retrospective and prospective studies that the response to radiotherapy and the prognosis, especially in cervical cancers, might be improved if the Hb levels during radiotherapy can be maintained in the optimal range, either by transfusions or by erythropoietin. The effect of any antianemic therapy should be analyzed according to whether or not treatment was successful with regard to achieving optimal Hb levels during irrradiation. Erythropoietin is probably more effective in steadily increasing and stabilizing Hb levels, but bears the risk of overcorrection of Hb levels. The clinical relevance of erythropoietin receptors on tumor cells remains questionable. CONCLUSIONS: Treatment of anemia with the objective of improving local control and survival in radiotherapy patients is probably more difficult and sophisticated than coping with symptoms of anemia or improving quality of life. Nevertheless, the potential of antianemic treatment is high on the basis of experimental and clinical data, and further clinical trials are warranted. PMID- 15549185 TI - Ductal carcinoma in situ of the breast. Increasing importance of radiotherapy as a part of the therapy approach. AB - BACKGROUND: The treatment of ductal carcinoma in situ of the breast (DCIS) has changed dramatically during the last decade. MATERIAL AND METHODS: In a review article, prospective randomized and retrospective studies of different treatment options of DCIS are evaluated to define the new role of radiotherapy. RESULTS: Until a few years ago, total mastectomy was the standard treatment of DCIS achieving a 95-98% cure rate. Three randomized studies show that adjuvant radiotherapy following local excision significantly reduces the rate of invasive and noninvasive recurrences by 40-60%, thus making breast conservation possible. Retrospective studies evaluated prognostic factors (tumor size; age; margin width, presence of necrosis; grading) in order to define subgroups of patients whom adjuvant radiotherapy can be safely spared. Adequately excised tumors with favorable grading and not associated with necrosis are potential candidates. Further evaluation in prospective randomized studies is, however, required. CONCLUSIONS: Radiotherapy is an essential part of breast-conserving treatment of DCIS to achieve sufficient local control. PMID- 15549186 TI - Internal mammary nodes in invasive breast carcinoma. To treat or not to treat? AB - BACKGROUND AND PURPOSE: According to recent data medial location of early breast cancer was associated with a higher risk of systemic relapse and breast cancer death compared with lateral location. This paper will focus on literature data and will also present own data on the prognostic impact of radiation therapy (RT) to internal mammary nodes (IMNs) in early breast cancer patients with medial hemisphere tumor location. PATIENTS AND METHODS: Four large recent series of the literature reporting on > 50,000 patients with special focus on the impact of tumor location were evaluated. No systematic RT to IMNs was applied. At the Department of Radiation Oncology, University of Erlangen, Germany, a total of 822 patients (492 with lateral and 330 with medial lesions) with early breast cancer were treated by surgery and postoperative RT with or without chemotherapy (1985 1996). All patients with medial lesions received RT to IMNs by a mixed-beam approach (50% photons, 50% electrons) with a total dose of 50 Gy. In patients with lateral lesions RT was directed to the breast alone (50.4 Gy total dose, boost 12-16 Gy). RESULTS: The magnitude of the negative impact of medial tumor location was relatively similar in all four studies available. According to Zucali et al., the hazard ratio (HR) for distant metastases was 1.29. The HR for breast cancer-specific survival (BCSS) varied between 1.46 (Lohrisch et al.) and 1.31 (Gaffney et al.). 5-year systemic disease-free survival (SDFS) rates were 66.3% and 74.2% for high-risk medial and lateral lesions, respectively (p < 0.005). Corresponding BCSS were 75.7% and 80.8%, respectively (p < 0.03, Lohrisch et al.). For all 822 patients following treatment on IMNs, 5-year overall survival (OS) for lateral lesions and medial lesions was 76.2% and 79.1% (n.s.), and SDFS for lateral and medial tumors 72.6% and 72.9% (n.s.), respectively. No subgroup could be identified in which prognosis of patients with medial tumors was inferior to survival data for patients with lateral lesions. In postmenopausal women, OS was significantly better for patients with medial versus lateral tumors (77.6% vs. 72.7%; p = 0.05); in patients receiving adjuvant chemotherapy SDFS (5 years) was better for those with medial versus lateral tumors (80.5% vs. 67.6%; p = 0.02). CONCLUSION: Consistent literature data exist indicating a diminished survival in patients with inner versus outer quadrant breast cancer. According to our data, RT with a total dose of 50 Gy to IMNs in breast cancer patients with medial lesions was associated with OS and SDFS rates comparable to patients with lateral tumors. PMID- 15549187 TI - Neoadjuvant radiochemotherapy in locally advanced gastric carcinoma. AB - BACKGROUND AND PURPOSE: Gastric carcinoma is characterized by a high rate of local recurrences and distant metastases and is often not resectable due to locally advanced stage. The aim of this study was to examine feasibility and effectiveness of neoadjuvant radiochemotherapy (RCT) for locally advanced, primarily nonresectable gastric carcinoma and to achieve curative resection. PATIENTS AND METHODS: 21 patients with locally advanced gastric cancer located in cardia (n = 17) and corpus (n = 4; seven cT3; 14 cT4; 18 cN+; all cM0) with a median age of 61 years were scheduled to receive neoadjuvant RCT. Therapy consisted of a conventionally fractionated, conformal radiotherapy using the shrinking-field technique (1.8 Gy to 45 Gy + 5.4 Gy) and chemotherapy using cisplatin (20 mg/m(2), d1-5, 29-33), 5-fluorouracil (5-FU; 800 mg/m(2), d1-5, 29 33) or paclitaxel (135 mg/m(2), d1, 29). 4-6 weeks after completion of RCT, surgery was performed whenever feasible. RESULTS: Hematologic toxicity was moderate with grade 3 leukopenia in 10/21 patients and grade 3 thrombopenia in 5/21 (CTC). Nonhematologic toxicities consisted of 5/21 cases of fever as well as one fungal sepsis. Following RCT, tumors were classified resectable in 16/21 patients (76%); 12/21 patients (58%) were operated on, 11/12 achieved clear margins (R0). Response was as follows: complete remission (CR) 3/21 (14%), partial remission 13/21 (62%), no change 3/21 (14%), systemic progressive disease (PD) 2/21 (10%). The median survival and the 2-year survival rates were 18 months and 42%, respectively, for the patients following R0 resections as compared to 10 months and 0% for the remaining patients (p = 0.035). Local control (4 years) for patients following R0 resection was 89%. CONCLUSION: Neoadjuvant RCT is feasible and locally highly effective but must be further investigated involving a higher number of patients. PMID- 15549188 TI - Current status of radiation therapy and combined-modality treatment for bladder cancer. AB - BACKGROUND: Standard treatment for muscle-invasive bladder cancer is radical cystectomy. Combined-modality treatment (CMT), including transurethral resection (TURBT), radiation therapy (RT) and systemic chemotherapy, has been shown to produce survival rates comparable to those of radical cystectomy. With these programs, cystectomy has been reserved for patients with incomplete response or local relapse after trimodality treatment. METHODS: This review summarizes series of radical RT with different fractionation schedules and focuses on CMT for muscle-invasive bladder cancer. Current protocols of the bladder-sparing approach will be discussed and the background of future developments, including incorporation of promising new chemotherapeutic agents as well as the role of predictive and prognostic factors in selecting patients for the respective treatment alternatives, will be given. RESULTS: There is moderate evidence that hyperfractionated and accelerated regimens are superior to conventional RT at least in situations where no concomitant chemotherapy is applied. Several phase II studies and one phase III study indicate that concomitant radiochemotherapy is superior to RT alone. In modern series of CMT, 5-year survival rates in the range of 50-60% have been published, and about three quarters of the surviving patients maintained their own bladder. Recent data suggest that incorporation of newer chemotherapeutic agents, particularly gemcitabine and taxanes, in CMT protocols is feasible and promising. Clinical criteria helpful in determining patients for bladder preservation include such variables as early tumor stage, unifocal tumor, a visibly and microscopically complete TURBT, and absence of ureteral obstruction. CONCLUSION: CMT for bladder cancer is a reasonable treatment option for patients who are deemed medically unfit for cystectomy and for those seeking an alternative to radical cystectomy. PMID- 15549189 TI - Treatment of oral cavity and oropharyngeal cancer. Indications, technical aspects, and results of interstitial brachytherapy. AB - Excellent local control rates of interstitial brachytherapy in oral cavity cancer and oropharyngeal carcinoma have been demonstrated in different retrospective studies. Compared to external-beam radiation therapy the high local control rates with a low rate of side effects obtained by interstitial brachytherapy are the result of a steep dose reduction in the implant-surrounding normal tissues. Therefore, interstitial brachytherapy offers rather the possibility to give high doses without inevitably leading to high complication rates. Low-dose-rate (LDR) and pulsed-dose-rate (PDR) interstitial brachytherapy with 0.4-0.55 Gy/h/24 h for tumors of the oral cavity and oropharynx in selected patients is a proven, effective and safe treatment method with excellent long-term data both as a sole treatment modality and a postoperative method, as well as a unique treatment method of head and neck tumors in previously irradiated areas. This paper deals with the technical aspects of interstitial brachytherapy, that seem to be relevant to high-quality outcome, and gives an overview of indications as well as past and recent results of interstitial brachytherapy in head and neck cancer. PMID- 15549190 TI - Radiation therapy for nonmalignant diseases in Germany. Current concepts and future perspectives. AB - BACKGROUND: Radiotherapy (RT) of nonmalignant diseases has a long-standing tradition in Germany. Over the past decade significant theoretical and clinical progress has been made in this field to be internationally recognized as an important segment of clinical RT. This development is reflected in a national patterns-of-care study (PCS) conducted during the years 2001-2002. MATERIAL AND METHODS: In 2001 and 2002, a questionnaire was mailed to all RT facilities in Germany to assess equipment, patient accrual, RT indications, and treatment concepts. 146 of 180 institutions (81%) returned all requested data: 23 university hospitals (UNI), 95 community hospitals (COM), and 28 private institutions (PRIV). The specific diseases treated at each institution and the RT concepts were analyzed for frequencies and ratios between the different institution types. All data were compared to the first PCS in 1994-1996. RESULTS: In 137 institutions (94%) 415 megavoltage units (mean 1.7; range 1-4), and in 78 institutions (53%) 112 orthovoltage units (mean 1.1; range 0-2) were available. A mean of 37,410 patients were treated per year in all institutions: 503 (1.3%) for inflammatory disorders, 23,752 (63.5%) for degenerative, 1,252 (3.3%) for hypertrophic, and 11,051 (29.5%) for functional, other and unspecified disorders. In comparison to the first PCS there was a significant increase of patients per year (from 20,082 to 37,410; +86.3%) in most nonmalignant diseases during the past 7-8 years. Most disorders were treated in accordance with the national consensus guidelines: the prescribed dose concepts (single and total doses) varied much less during the period 2001-2002 in comparison with the previous PCS in 1994-1996. Only five institutions (3.4%) received recommendations to change single or total doses and/or treatment delivery. Univariate analysis detected significant institutional differences in the use of RT for various disorders. CONCLUSION: RT is increasingly accepted in Germany as a reasonable treatment option for many nonmalignant diseases. The long-term perspective and research plan will have to include various updates of PCS, re-writing of consensus guidelines, introduction of registries for rare nonmalignant disorders, and clinical controlled studies even for so-called established indications, as international acceptance is based on the criteria of evidence-based medicine. PMID- 15549191 TI - Radiation sensitization by inhibition of activated Ras. AB - BACKGROUND AND PURPOSE: Ras has been identified as a significant contributor to radiation resistance. This article reviews preclinical and phase I clinical studies that reported on combining inhibition of activated Ras and downstream effectors of Ras with radiotherapy. MATERIAL AND METHODS: Transfection studies and RNA interference were used to check the role of the Ras isoforms for intrinsic radiation sensibility. Western blotting was used to control for prenylation inhibition of the respective Ras isoforms and for changes in activity of downstream proteins. Clonogenic assays with human and rodent tumor cell lines served for testing radiosensitivity. In vivo, farnesyltransferase inhibitors (FTIs) and irradiation were used to treat xenograft tumors. Ex vivo plating efficiency measurements, regrowth of tumors, and EF5 staining for detection of hypoxia were endpoints in these studies. Simultaneous treatment with L-778,123 and irradiation was performed in non-small cell lung cancer, head and neck cancer, and pancreatic cancer patients. RESULTS: Radiation sensitization was achieved in vitro and in vivo blocking the prenylation of Ras proteins in cell lines with Ras activated by mutations or receptor signaling. Among the many Ras downstream pathways the phosphoinositide 3 (PI3) kinase-Akt pathway was identified as a contributor to Ras-mediated radiation resistance. Furthermore, increased oxygenation was observed in xenograft tumors after FTI treatment. Combined treatment in a phase I study was safe and effective. CONCLUSION: The rational combination of FTIs with radiotherapy may improve the clinical results of patients with tumors who bear mutant or receptor-signaling activated Ras. PMID- 15549194 TI - [Epidemiological studies on the health state of the population]. PMID- 15549195 TI - [Methodological views on the SF-36 summary scores based on the adult German population]. AB - The SF-36 is a widely used instrument to measure health-related quality of life that provides a profile of eight scales [Physical Functioning (PF), Role-Physical (RP), Bodily Pain (BP), General Health (GH), Vitality (VT), Social Functioning (SF), Role-Emotional (RE) and Mental Health (MH)]. To reduce the dimension of the profile, a principal component analysis with the data of the US SF-36 Survey was undertaken. As a result, the Physical Component Summary (PCS) and the Mental Component Summary (MCS) were gained by weighting and summing up of the original scales of the SF-36. The weights were assumed to be country specific. Comparing the weights from the representative samples of nine European countries with those from the USA it has been suggested that the weights gained from the US sample could be applied to all datasets for purposes of comparability and simplicity. In this paper, for a new representative German population sample completed in 1998, the calculation results are compared with those for the older German population sample from 1994. PCS and MCS are calculated using the weights from the new German population sample as well as using the American weights. In this paper, the calculations for the 1998 representative population sample were repeated based on the US weights developed in 1994. The German weights were also calculated based on the 1998 sample. The results were compared and the pro and cons of using the American weights are discussed. In essence, the American weights continue to be acceptable for international comparisons. The separate evaluation of physical and mental health for men and women allows additional insight into gender-specific changes. In this paper, the summary scales for the 1998 German normative population are calculated gender specific. The weights to be used in future studies are also published. With respect to gender mainstreaming, this approach should be taken into consideration in quality of life research. Regarding the methodological and factorial difficulties, the questions arise if the summary scales should be applied instead of the original scales of the SF-36 questionnaire. PMID- 15549196 TI - [Social inequality and smoking behavior in Germany]. AB - Data of the National Health Survey 1998 are used to analyze the impact of alternative indicators of social inequality on smoking of the 18- to 79-year-old population in Germany. The members of socially disadvantaged groups not only smoke more frequently, they show also lower rates in smoking cessation. This is most obvious in middle age while only small differences can be observed at older ages. Multivariate analyses evidence primarily education as a determinant of smoking among men and women. The effects of income and occupational status are fairly small. The results point out that health inequalities can partly be explained by different smoking habits and, therefore, underline the necessity of a differentiated and group-specific tobacco prevention and tobacco control policy. PMID- 15549197 TI - [Self-medication with over-the-counter (OTC) preparations in Germany]. AB - Self-medication with OTC drugs is attracting attention in developed countries. This study examines prevalence and determinants of OTC drug use in a representative sample of German adults aged 18-79 years. A total of 7099 participants of the Drug Utilisation Survey were interviewed regarding drug use including OTC use within the last 7 days prior to the interview. This survey is a part of the representative German National Health Interview and Examination Survey 1998. Complete information is available from 3393 men and 3594 women: 17.6% of men and 10.8% of women use self-medicated OTC drugs exclusively, whereas 12.3% and 29.3%, respectively, use OTC drugs in addition to prescribed drugs. Besides sex, factors such as age, socioeconomic status and community size determine self-medication with OTC drugs. The most commonly used self-medicated OTC drugs are vitamins, minerals and analgesics. The most commonly reported indication for self-medication with OTC drugs is "prevention". Beside prescribed medication, self-medication with OTC drugs is an important part of drug usage for men and women in Germany. Health consciousness appears to be a significant factor to explain the prevalence OTC drug use. PMID- 15549198 TI - [Drug usage of men and women with coronary heart disease. Results of the German Federal Health Survey 1998]. AB - When managing the risk factors of coronary heart diseases, therapies using drugs play an important role. This present study analyses self-reported data on current drug usage of men and women with coronary heart disease. The data were collected as a part of a wider drug utilisation survey which in turn is a module of the German National Health Interview and Examination Survey 1998 (Bundes Gesundheitssurvey 1998). This survey was compiled by conducting a standardised computer-assisted medical interview with a representative sample of the German population between the ages of 18 and 79 years. Of the 7099 participants, 209 of the women and 252 of the men affirmed having had a medically diagnosed coronary heart disease (CHD) such as angina pectoris and/ or myocardial infarction. Slightly more men (87.3%) were using drugs to treat CHDs than the women (86.1%). They (the men) also used more drugs on average (men 3.3, women 3.0). These differences however were not statistically significant. Clear and statistically significant differences were present in the usage of salicylic acid (men 54.4%, women 45.6%), HMG CoA reductase inhibitors (men 26%, women 15.3%) and cardiac glycosides (men 14%, women 25.8%). The higher usage prevalence rate of salicylic acid found in the men still remained after taking their age, social status and the region (east vs west) into account. The differences in the usage of specific drugs in the treatment of CHDs in men and women indicate a difference in prescription behaviour in the ambulatory medical care. Whether and to which extent gender-specific differences still remain today has to be decided by using current epidemiological data representative of the population. PMID- 15549199 TI - [Use of vitamin and mineral supplements in Germany]. AB - Recent regulations on supplement use in Europe address the necessity to include information on dietary intake to derive maximum amounts for vitamins and minerals in food supplements. Therefore, information about supplement use in Germany, as well as data on micronutrient intake (from supplements and traditional foods), in particular of persons who use several supplements, is examined in detail. As part of the representative German National Health Interview and Examination Study 1998, in the Nutrition Survey 4030 persons, aged 18-79 years, were asked about their dietary habits, including vitamin and mineral supplement use, in a personal computer-aided interview. About 43% of the population reported using supplements at least once in the observation period of 12 months. Women use supplements more frequently than men. A relatively small group uses several supplements, most often vitamin plus multivitamin supplements. Considering single nutrients, vitamin C is most commonly consumed from more than one supplement. However, a micronutrient intake above the tolerable upper intake level is rare. Use of supplements is common in Germany. Nevertheless, only a small proportion of the population uses more than one supplement containing the same micronutrient daily. Even among these persons an intake above the tolerable upper intake level is seldom. PMID- 15549200 TI - [Environmental Survey for Children-- the environmental module of KiGGS. I. Design and research program]. AB - The German Environmental Survey for Children (GerES IV) is the environment oriented module of the National Health Interview and Examination Survey for Children and Adolescents (KiGGS) which is being performed nationwide in Germany. From 2003 to 2006, a random subsample of 1800 children aged 3-14 years is being studied with regard to their body burden and health impairments linked to housing conditions and the personal environment- and health-relevant behaviour. The basic study programme includes the analysis of blood, urine, tap water and house dust as well as the application of an extensive questionnaire. The data gained from this population sample, which is representative for Germany's children, are the basis for deriving reference values to characterise the background exposure of children aged 3-14 years. Trends over time can be detected and the success of environmental policies verified by comparing the data with those of the German Environmental Survey 1990/92 (GerES II), also conducted in close cooperation with the National Health Survey, which included children aged 6-14 years. By linking the data from the Environmental and the Health Surveys, health-relevant environmental exposures can be detected and different scientific hypotheses can be tested. The main subjects that are being dealt with using subcollectives of GerES IV are 'VOC and eye and nasopharynx irritation', 'indoor allergens and allergic diseases of the respiratory system', 'chromium, nickel, fragrances and contact allergens', and 'noise, hearing capacity and stress hormones'. PMID- 15549202 TI - [Comparison of mycological and chemical analytical laboratory methods for detecting mold damage in indoor environments]. AB - To evaluate frequently used methods that discriminate between moldy and nonmoldy indoor environments, 45 homes with visible mold growth and 47 definitively non infested homes, both confirmed by inspection, were investigated by microbiological and chemical analytical methods. The study was laboratory blinded in relation to the confirmed mold status of the rooms. Statistical evaluation of the results of the applied mycological methods with the Receiver Operating Characteristic (ROC) curve analysis showed that these methods (impaction, open Petri dish method, and determination of mold spores in house dust samples) performed very well in discriminating between rooms with visible mold growth and nonmoldy rooms when the sum score of the mold genera Aspergillus and Penicillium was used as an indicator. The calculated areas under the ROC curves (AUC) of the three mycological methods were: 0.992 (95% CI 0.942-0.997) for mold spores in house dust samples, 0.996 (95% CI 0.940-0.998) for the open Petri dish method, and 0.999 (95% CI 0.957-1.000) for the determination of airborne spores with the Andersen impactor, respectively. A perfect discrimination would lead to an AUC of 1. These results were obtained with DG 18-agar as well as with malt extract agar. In contrast to the results of the mycological methods, the chemical analytical method under the same study conditions showed a distinctly lower performance in discriminating rooms according to their mold status when a sum score (concentration of eight typical MVOC) was used as an indicator. The area under the ROC curve (AUC) had a value of 0.620 (95% CI 0.509-0.723). A completely useless test would have an AUC of 0.5. As the lower limit of the 95% confidence interval of the area under the ROC curve is close to 0.5, the results obtained with the MVOC method do not differ from the classification results which can be obtained simply by chance. Possible methodological biases which could have lead to this interpretation are discussed. PMID- 15549201 TI - [German Environmental Survey for Children (GerES IV). Environmental module of KiGGS. Part 2: The first year of field work]. AB - The field work of the German Environmental Survey for Children (GerES IV) was started nationwide in May 2003. The survey is a module of the National Health Interview and Examination Survey for Children and Adolescents (KiGGS). This environmental survey includes a representative subsample of 1800 children from the 18,000 participants of the KiGGS who are being examined regarding health relevant environmental exposure. The investigational programme of the survey was tested successfully in a pilot study (pretest) the results of which contributed to the optimisation of the main study. The field work is essentially done by the environmental interviewer of the three teams of the KiGGS. Preparation, organization and accompaniment of the work is done by the coordination centre of the Robert Koch Institute on Federal Environmental Agency (UBA) instructions. Funding agencies are the Federal Ministry of Education and Research (BMBF) and the Federal Ministry for the Environment, Nature Conservation and Nuclear Safety (BMU). Both the KiGGS and the environmental module are evaluated by internal and external quality assurance during the whole study period (May 2003-May 2006). Up to now the established teams have done their field work very well and the coordination of the overall project by the Robert Koch Institute works well, too. A total of 722 subjects had taken part in the GerES IV by July 2004. PMID- 15549203 TI - [Suicidal methods--a comparison between East and West Germany. Epidemiological, forensic and sociomedical aspects]. AB - A comparative study of the means of suicide in West and East Germany is extremely difficult both because of the data stock and the methodological questions that arise. This examination analyses means of suicide used during the following years: 1962-1964, 1987-1989 and 2000-2002. Accordingly, there is a similar pattern in both East and West for type of suicide committed and distinct developmental parallelism in applying the suicide methods. Certain means of suicide are more prominent (e. g. "hanging", "falling from a height") while others decrease in significance (e. g. "drowning"). No link was found between means of suicide and "society order". The use of a certain means of suicide depends on "availability", "existence" or "accessibility" of opportunities to commit suicide in the everyday environment. The same trend towards an increase of "hard" means of suicide was noted. The ways in which suicide is committed are becoming increasingly alike, but in the East it is apparent that people adhere more to "traditional" means of suicide. PMID- 15549204 TI - [23rd Jena Symposium: Campylobacter infections]. PMID- 15549205 TI - [Report on the 67th and 68th of the provisional Commission for Cosmetic Medication of the Federal Institute for Risk Assessment (BfR) from 3 and 4 December 2003 and 6 May 2004 in Berlin]. PMID- 15549206 TI - [RKI-Commission "Methods and Quality Assurance in Environmental Medicine". Announcement from the Commission]. PMID- 15549207 TI - [Genetic polymorphism (sequence variation) of foreign substance-metabolizing enzymes and their significance in environmental medicine. Statement of the Commission "Methods and Quality Assurance in Environmental Medicine"]. PMID- 15549208 TI - [Suspension of adequate disinfection capacity (section sign) 5 Abs. 4 TrinkwV 2001 for exceptional incidents or emergencies. Statement of the Federal Environmental Agency after a hearing of the Drinking Water Commission of the Federal Ministry for Health and Social Security for the Federal Environmental Agency]. PMID- 15549209 TI - [New aspects in uv and photochemotherapy]. AB - Recent data show that from a pharmacological point of view topical (cream or bath) PUVA therapy is superior to systemic PUVA. Due to a significant reduction of side effects compared to systemic PUVA, bath PUVA has now started to replace oral PUVA therapy. Narrowband UVB has proved to be superior to broadband UVB in the treatment of psoriasis and is effective for a number of dermatoses such as vitilgo, atopic dermatitis and polymorphic light eruption. UVA1 phototherapy is highly effective in the treatment of moderate to severe atopic dermatitis and sclerosing diseases of the skin. Data dealing with UVA1 phototherapy for other indications are still preliminary. High-dose UVA1 is has been widely replaced by medium-dose UVA1, as a number of studies have shown similar therapeutic efficacy of both dose regimens. PMID- 15549210 TI - [Diagnostics of the cochlear amplifier by means of DPOAE growth functions]. AB - Extrapolated DPOAE growth functions can be applied in ENT diagnostics for a specific assessment of cochlear dysfunction. In screening newborn hearing, they are able to detect transitory sound conductive hearing loss and thus help to reduce the rate of false positive TEOAE responses in the early postnatal period. Since DPOAE growth functions are correlated with loudness functions, DPOAEs offer the potential for basic hearing aid adjustment, especially in children. Extrapolated DPOAE I/O-functions provide a tool for a fast, automated frequency specific and quantitative evaluation of hearing loss. However, DPOAE diagnostics is limited to a hearing loss of 50 dB HL. Thus, a combined measurement of DPOAE and AMFR would be useful. PMID- 15549211 TI - [Computer assisted methods in reconstructive and function-preserving orbital surgery. New capabilities of computer assisted preoperative surgical planning (CAPP) and computer assisted surgery (CAS)]. AB - BACKGROUND: Orbital reconstruction after tumor or trauma makes high demands on the surgeon when restoring the optic axis and cosmetic features. Computer assisted preoperative planning surgery (CAPP) and computer-assisted surgery (CAS) allow calculation of the form of the orbital cavity to be reconstructed preoperatively as well as the process of its realization intraoperatively. METHODS: We developed new planning software methods for this surgical procedure. For validation the deviation of accuracy was assessed between the virtual and the real model in eight patients with surgical reconstruction of the orbit. RESULTS: The check of accuracy of the reconstructions compared to the planning of the surgery by fusion of preoperative and postoperative data resulted in a mean deviation of 1.74 mm. The volumetric measurements presented a mean deviation of 0.2 cm(3) with an accuracy of >99%. The accuracy of linear measurements with the techniques of image fusion of the "feature-based" method was 0.38 mm and thus close to the technical threshold of effective data accuracy of 0.35 mm. The enophthalmus could be assessed more objectively with this method compared to using the Hertel index. CONCLUSIONS: CAPP and CAS are helpful in reconstructive orbital surgery to verify the position of the optic axis and the cosmetic result intraoperatively. PMID- 15549212 TI - [Diagnostic and therapeutic procedure for spontaneous emphysema of the neck and mediastinum]. AB - BACKGROUND: Emphysema without any etiological indices from the history represents a diagnostic and therapeutic challenge. PATIENT COLLECTIVE: Over the last 5 years, we treated four patients (three male, one female; aged 3-29 years) with cervical and/or mediastinal emphysema of unknown cause. RESULTS: Two young men with cervical emphysema were observed and received prophylactic antibiotic treatment. After involution of the emphysema, we performed an endoscopy which revealed no abnormalities. A female patient and a 3-year-old boy had a history of coughing and a query history of foreign body ingestion before the appearance of the emphysema. The immediate endoscopies were without pathologic findings. All patients recovered completely without any complications or recurrences. CONCLUSIONS: If there is no indication for a foreign body or a trauma in the history or in radiological imaging, endoscopy of the airways and the upper digestive tract should follow when the emphysema has subsided. The aim is to avoid any further spread of the emphysema and of pathogens. If there is a history of a foreign body or trauma, an immediate endoscopy is indicated. PMID- 15549213 TI - [Electronic surveillance of nosocomial infections in head and neck surgery and stapes surgery]. AB - BACKGROUND: Nosocomial infection is generally regarded as the most important postoperative complication. Therefore, on 28 December 2002 a German law was implemented requiring the surveillance of postoperative infections in all hospitals. METHODS: The authors propose using stapes and thyroid surgery to monitor the infection rate in a typical Head and Neck Department. A versatile software (CISS) based on MS Word and MS Excel was developed for this purpose. RESULTS: Postoperative infections were retrospectively analyzed for three subsequent years. The present data confirm the results of previous studies that surveillance itself is a powerful tool in reducing the postoperative infection rate. CONCLUSION: The newly developed software provided an easy tool for the collection of infection data. The reported infection rates in stapes and thyroid surgery are representative of ENT clinics in Germany. PMID- 15549214 TI - [Magnetic Drug Targeting--a new approach in locoregional tumor therapy with chemotherapeutic agents. Experimental animal studies]. AB - BACKGROUND: Advanced squamous cell carcinomas of the head and neck region were often treated with combined radio-chemotherapy. Radiotherapy allows a focused treatment of the tumor, and healthy tissue can be protected from radiation. Chemotherapy, however, is mostly given systemically and the unwanted negative side effects also develop in many other organs. AIM OF THE STUDY: Locoregional application of chemotherapeutic agents with Magnetic Drug Targeting on an animal experimental study. METHODS AND RESULTS: Magnetic Drug Targeting is a new approach to the locoregional treatment of tumors. Ferrofluids (colloidal dispersion of magnetic nanoparticles) were reversibly bound to chemotherapeutic agents and injected intra-arterially, while focused with an external magnetic field to a certain body compartment (i.e. the tumor). With only 20% or 50% percent of the regular systemic chemotherapeutic dose, we achieved an up to 26 times higher concentration in the tumor region with this application compared to the usual systemic administration. CONCLUSION: Magnetic Drug Targeting offers an unique opportunity to treat tumors locoregionally with chemotherapeutic agents. PMID- 15549215 TI - [Endoscopy of the upper gastrointestinal tract]. AB - Esophagogastroduodenoscopy (EGD) has replaced X-ray diagnosis as the standard method for assessment of the upper gastrointestinal tract. It also offers an array of minimally invasive treatment options. This contribution presents the requisite medical, technical, and human resources needed for EGD. The indication for this invasive procedure should always be carefully reviewed and contraindications excluded. EGD should be performed according to standardized procedures and well documented by noting distinctive features of peristalsis and describing alterations of mucosal size and surface texture. The endoscopic techniques of biopsy, chromoendoscopy, and percutaneous endoscopic gastrostomy should always be available in routine endoscopy. PMID- 15549216 TI - [Pharmacological treatment in adults with attention deficit hyperactivity disorder]. AB - Treatment in adults with attention deficit hyperactivity disorder predominantly relies on pharmacotherapeutic approaches especially with psychostimulants. Empirical studies indicate that their clinical effectiveness may be as high as in children and adolescents, especially in higher dosages. However, due to the high prevalence of comorbidities, e.g. depression, psychopharmacological treatment requires an extended use of other substance groups, especially antidepressants. An optimal treatment response necessitates the choice of an adequate substance depending on the leading clinical symptoms and a procedure of an individual titration of different dosages. This article reviews the current empirical results in the pharmacological treatment of ADHD in adults and provides possible treatment strategies for clinical practice. PMID- 15549217 TI - [Regional and local clusters of railway suicides]. AB - BACKGROUND AND METHOD: Suicide research has identified regional and local clusters of increased suicide prevalence. As part of a comprehensive prevention strategy to fight railway suicides, we sought to identify such particular clusters on the German Railway net (total length 37,080 km) during a 6-year observation period (1997-2002). Data stem from the central registry of all person accidents on the German Railway net. RESULTS: During the observation period, a total of 5,731 suicides (in average 18 cases per week) were registered. Against expectation derived from distribution patterns in the general suicide research, we found a preponderance in the southern part compared to the northern part of Germany and a significant downward trend from west to east ( p=0.004). Most suicides occurred on open track (66%) compared to suicides at railway platforms (34%). We identified 16 places of high risk with 6 to 29 suicides per railway km. Of these high risk places, 75% were in the proximity of psychiatric hospitals. CONCLUSION: Increased awareness for regional and local suicide clusters, initiatives to reduce the accessibility and warning infra-red beams are among recommendations for suicide prevention derived from these data. PMID- 15549220 TI - [Significance of Doppler ultrasound procedures for diagnosis of carotid stenoses]. AB - Determining degree and morphology of stenoses is important for surgical planning or stent implantation. Vascular ultrasound is usually the first modality to evaluate carotid artery stenosis. Due to rapid development various methods of vascular ultrasound are applied including continuous wave (CW) Doppler, duplex Doppler, colour-coded duplex sonography (CCDS), power Doppler and B-flow technique. For quantitative assessment of the degree of stenosis the most frequently used parameters are peak systolic velocity (PSV), end-diastolic velocity (EDV) in the internal carotid artery (ICA), as well as ICA to CCA ratios of PSV and EDV. Different results reported in the literature may reflect differences in defining the degree of stenosis and methodological differences in protocol or imaging techniques. Differences in defining the degree of stenosis, advantages and disadvantages of the different Doppler techniques and future developments are discussed in detail. PMID- 15549221 TI - [The significance of MR angiography for the diagnosis of carotid stenoses]. AB - Arteriosclerosis of the supra-aortic vessels is a known risk factor for cerebrovascular disease. Large multicenter studies like NASCET and ECST have proven the protective effects of the surgical treatment of severe (>70%) carotid artery stenoses. The exact detection and grading of stenoses of the carotid arteries is a prerequisite for a successful therapy. Besides the gold-standard examination, digital subtraction angiography (DSA), MR angiography (MRA) is being used increasingly. MRA allows a non-invasive, high-resolution depiction of the supra-aortic vessels without ionizing radiation in only 20 s. This article first illustrates the MRA-techniques, and the indications and limitations of MRA. Next, the diagnostic accuracy of MRA is compared to DSA and ultrasound. The clinical value of MRA for the detection of stenoses of the carotid arteries is discussed on the basis of clinical examples. PMID- 15549222 TI - [Distal protection devices in carotid stent]. AB - Endovascular treatment of carotid stenoses is increasingly used. Frequently temporary occlusion balloons or filters are placed distal to the stenosis to gain additional protection against cerebral emboli. There is still a scientific debate about the usefulness of such distal protection devices. The following contribution reviews existing techniques and their clinical results and should allow for a critical discussion and selection of current techniques and materials. PMID- 15549223 TI - [Unilateral eyelid swelling after a simple nose blowing]. PMID- 15549224 TI - [Surgical requirements for radiological diagnostics of liver pathologies]. AB - BACKGROUND AND PURPOSE: Radiology is an essential preoperative tool for a liver surgeon to plan extent of resection and potential difficulties during liver surgery. MATERIAL AND METHODS: Primary goal in defining liver pathologies is a careful patients' history, a clinical evaluation and reviewing at least one radiological film one could acquire. Don't rely on written reports that may direct you in a useless track. This overview tries to address the essential radiological requests of a surgeon in defining liver tumors ethiology and best optional treatment. RESULTS: Major advances in radiologic diagnostics led to an improvement in the adequate staging of a given liver pathology. Therefore we are nowadays able to inform our patients about possible treatment options without leaving a big gap to possible intra-operative findings which may alter the therapy. Surgical exploration to define therapeutic strategies becomes fundamental only in a minority of patients with unclear preoperative imaging studies. DISCUSSION: Interdisciplinary groups should define future strategies in a patient with a given liver pathology. Specialisation has defined the hepatobiliary surgeon which should be consulted in case of a liver or biliary tumor to guide possible therapeutic treatment options. PMID- 15549225 TI - [(1)H magnetic resonance spectroscopy (MRS) of the liver and hepatic malignant tumors at 3.0 Tesla]. AB - Use of whole-body MRI beyond 1.5 Tesla (T) has initiated a renaissance in spectroscopic procedures (MRS). The superior signal-to-noise ratio of clinical 3T tomographs allows reliable acquisition of MR spectra not only in fixed organs but also in targets moved by breathing such as the liver. The following contribution describes the principles of (1)H MRS and our own initial experiences with spectroscopy of the liver and hepatic malignant tumors with 3T whole-body MRI. PMID- 15549226 TI - [Current status of MRI diagnostics with liver-specific contrast agents. Gd-EOB DTPA and Gd-BOPTA]. AB - The contrast agents Gd-EOB-DTPA and Gd-BOPTA can be administered by bolus injection and are appropriate for use in MRI both as vascularization markers and markers of hepatobiliary excretion. This contribution presents an overview of the specific characteristics of contrast media and the status of clinical development. In comparison to CT and to MRI with unspecific extracellular Gd chelates, liver-specific contrast agents offer advantages in differentiating unclear liver lesions, increasing the detection rate, and examining the bile duct system. PMID- 15549227 TI - [Modern visualization of the liver with MRT. Current trends and future perspectives]. AB - This contribution provides an overview and imparts basic knowledge on pertinent technical developments in magnetic resonance imaging (MRI) of the liver: 3D sequences, respiratory triggering, parallel imaging, and 3 Tesla (3T). 3D sequences can be used as T1-weighted (T1w) sequences for analyzing dynamics of contrast enhancement or as T2w sequences for MR cholangiography. Consistent improvements in respiratory triggering make it possible to obtain good image quality on T2w scans even in patients unable to hold their breath. Parallel imaging as a universal technique to accelerate image acquisition is particularly appropriate for MRI of the liver, and it has been shown that the reduced acquisition time is not achieved at the expense of image quality. Further progress in MRI of the liver can be expected with use of the 3T systems, but hitherto irrelevant problems must still be solved. Overall the innovations presented here, applied alone or in combination, facilitate rapid, robust, and high-quality MRI diagnostic assessment of the liver. PMID- 15549229 TI - Analysis of QTLs for yield, yield components, and malting quality in a BC3-DH population of spring barley. AB - Advanced backcross (AB)-quantitative trait locus (QTL) analysis has been successfully applied for detecting and transferring QTLs from unadapted germplasm into elite breeding lines in various plant species. Here, we describe the application of a modified AB breeding scheme to spring barley. A BC3-doubled haploid (DH) population consisting of 181 lines derived from the German spring barley cultivar 'Brenda' (Hordeum vulgare subsp. vulgare) as the recurrent parent and the wild species line 'HS213' (H. vulgare subsp. spontaneum) as the donor line was evaluated for yield and its components as well as malting quality traits. A set of 60 microsatellite markers was used to genotype the population, and phenotypic data were collected at two locations in Germany in continuous years. Altogether, 25 significant QTLs were detected by single-marker regression analysis and interval mapping. Most positive QTLs originated from the recurrent parent 'Brenda'. A QTL, Qhd2.1, on chromosome 2HS from 'Brenda' explained 18.3% and 20.7% of the phenotypic variation for yield and heading date, respectively. Due to the small percentage of donor-parent genome of 6.25%, the BC3-DH lines could be directly used for the extraction of near-isogenic lines (NILs) for Qhd2.1. Consequently, it was possible to determine the precise location of the locus hd2.1 within a region of 6.5 cM, using an F2 population consisting of 234 individuals developed from a cross between an NIL containing a defined donor segment at this locus and 'Brenda'. The location of this QTL was consistent with the presence of a major photoperiod response gene, Ppd-H1, previously reported in this region, which is associated with pleiotropic effects on yield components. In summary, the analysis of a BC3-DH population in barley provides a compromise between the analysis of QTLs by means of an AB scheme and the generation of defined substitution lines. Several lines carrying defined different donor segments for only one single chromosome or trait in the genetic background of 'Brenda' could be selected for further genetic studies. PMID- 15549230 TI - Epistasis underlying female sterility detected in hybrid breakdown in a Japonica Indica cross of rice (Oryza sativa L.). AB - Epistasis is considered to be a primary genetic basis of hybrid breakdown. We found novel epistatic genes causing hybrid breakdown in an intraspecific cross of cultivated rice (Oryza sativa L.). F2 progeny derived from a cross between a Japonica variety, Asominori, and an Indica variety, IR24, showed segregation of high sterility for seeds, even though the reciprocal F1 hybrids showed about 60% seed fertility. Backcross populations (BC3F2, BC3F3), obtained from repeated backcrossing with Asominori, showed the segregation of causal genes in a simple Mendelian fashion. Using these populations, we identified that this sterility was hybrid breakdown caused by interaction among three nuclear genes distributed on the both parental genomes. These new genes, designated as hsa1, hsa2, and hsa3, were found to be involved in female gamete development by histological examination. The Indica parent IR24 has a sterile allele, hsa1-IR, which was located at near RFLP marker G148 on chromosome 12, whereas the Japonica parent Asominori has two sterile alleles, hsa2-As on chromosome 8 (close to G104) and hsa3-As on chromosome 9 (close to RM285). Female gametes carrying the hsa1-IR, hsa2-As, and hsa3-As alleles aborted in hsa1-IR homozygous plant, leading to seed sterility and selective elimination of the specific allelic combination. This study provides direct evidence that hybrid breakdown is attributed to epistatic interaction of genes from both parents and suggests that complicated mechanisms has been developed for hybrid breakdown during the evolution of rice. PMID- 15549231 TI - High-resolution genetic mapping at the Bph15 locus for brown planthopper resistance in rice (Oryza sativa L.). AB - Resistance to the brown planthopper (BPH), Nilaparvata lugens Stal, a devastating sucking insect pest of rice, is an important breeding objective in rice improvement programs. Bph15, one of the 17 major BPH resistance genes so far identified in both cultivated and wild rice, has been identified in an introgression line, B5, and mapped on chromosome 4 flanked by restriction fragment length polymorphism markers C820 and S11182. In order to pave the way for positional cloning of this gene, we have developed a high-resolution genetic map of Bph15 by positioning 21 DNA markers in the target chromosomal region. Mapping was based on a PCR-based screening of 9,472 F(2) individuals derived from a cross between RI93, a selected recombinant inbred line of B5 bearing the resistance gene Bph15, and a susceptible variety, Taichung Native 1, in order to identify recombinant plants within the Bph15 region. Recombinant F(2) individuals with the Bph15 genotype were determined by phenotype evaluation. Analysis of recombination events in the Bph15 region delimited the gene locus to an interval between markers RG1 and RG2 that co-segregated with the M1 marker. A genomic library of B5 was screened using these markers, and bacterial artificial chromosome clones spanning the Bph15 chromosome region were obtained. An assay of the recombinants using the sub-clones of these clones in combination with sequence analysis delimited the Bph15 gene to a genomic segment of approximately 47 kb. This result should serve as the basis for eventual isolation of the Bph15 resistance gene. PMID- 15549232 TI - Identification and characterization of a QTL on chromosome 2 for cytosolic glutamine synthetase content and panicle number in rice. AB - A quantitative trait locus (QTL) associated with the protein content of cytosolic glutamine synthetase (GS1; EC 6.3.1.2) in senescing leaves, panicle number, and panicle weight was characterized in rice (Oryza sativa L.). A near-isogenic line (NIL), C-22, developed by marker-assisted selection was grown under different nitrogen levels in the greenhouse and in a paddy field. Chromosome 2 of C-22 had an approximately 50-cM segment substituted from the Kasalath (indica) chromosome in a Koshihikari (japonica) genetic background. C-22 showed a 12-37% lower content of GS1 protein in leaf blades than Koshihikari, which was in good agreement with a QTL region positively affected by the japonica chromosome. At an early vegetative stage, C-22 had more active tillers than Koshihikari in the greenhouse. At the reproductive stage, both panicle number and total panicle weight of C-22 were significantly higher than those of Koshihikari, particularly when the plants were grown under a low-nitrogen condition. These traits of C-22 were further confirmed in a paddy field. Thus, tiller development was positively affected by the Kasalath chromosome at an early vegetative stage, which resulted in an increased panicle number and panicle weight at the mature stage in C-22. These data indicate that the target QTL (Pnn1; panicle number 1) is important in the development of tillers and panicles in rice. Linkage analyses for panicle number and ratio of developing tiller formation in the second axil (RDT) revealed that Pnn1 was delimited at the 6.7-cM region. PMID- 15549233 TI - Evaluation of cleaved amplified polymorphic sequence markers for Chamaecyparis obtusa based on expressed sequence tag information from Cryptomeria japonica. AB - We have developed and evaluated sequence-tagged site (STS) primers based on expressed sequence-tag information derived from sugi (Cryptomeria japonica) for use in hinoki (Chamaecyparis obtusa), a species that belongs to a different family (although it appears to be fairly closely related to sugi). Of the 417 C. japonica STS primer pairs we screened, 120 (approximately 30%) were transferable and provided specific PCR amplification products from 16 C. obtusa plus trees. We used haploid megagametophytes to investigate the homology of 80 STS fragments between C. obtusa and C. japonica and to identify orthologous loci. Nearly 90% of the fragments showed high (>70%) degrees of similarity between the species, and 35 STSs indicated homology to entries with the same putative function in a public DNA database. Of the 120 STS fragments amplified, 72 showed restriction fragment length polymorphisms; in addition, the CC2430 primers detected amplicon length polymorphism. We assessed the inheritance pattern of 27 cleaved amplified polymorphic sequence markers, using 20 individuals from the segregation population. All the markers analyzed were consistent with the marker inheritance patterns obtained from the screening panel, and no markers (except CC2716) showed significant (P<0.01) deviation from the expected segregation ratio. In total, 136 polymorphic markers were developed using C. japonica-based STS primers without any sequence modification. In addition, the applicability of STS-based markers developed in one species to other species was found to closely reflect the evolutionary distance between the species, which is roughly concordant with the difference between their rbcL sequences. We plan to use these markers for genetic studies in C. obtusa. Most of the markers should also provide reliable anchor loci for comparative mapping studies of the C. obtusa and C. japonica genomes. PMID- 15549235 TI - The complexity of non-enzymatic glycation product sets of human globins. AB - AIMS/HYPOTHESIS: Recently an individual variability in the relationships between mean blood glucose levels and HbA1c has been observed among diabetic patients. The aim of this study was to provide an accurate description and evaluation of glycated and glyco-oxidated globins from diabetic subjects and their relationship with HbA1c and plasma glucose values. METHODS: We studied 20 type 2 diabetic and 10 healthy subjects. Plasma samples were analysed by matrix-assisted laser desorption ionisation mass spectrometry. RESULTS: The presence of glycated and glyco-oxidated species of both alpha and beta globin was demonstrated. Values for these showed a good linear relationship with HbA1c values and the mean daily plasma glucose values for the 6 weeks preceding the investigation. Trends differed according to whether patients had chronic complications or not, differences being seen in the slopes of the plots relating HbA1c to the abundance of glycated and glyco-oxidated species. CONCLUSIONS/INTERPRETATION: The data obtained are consistent with the concept that individuals have a different individual proclivity for oxidation and/or that different oxidation kinetics are related to behavioural and environmental factors. Our data are thus relevant to the analysis of phenotype differences in diabetic patients. PMID- 15549236 TI - Epidemic theory and group violence. AB - BACKGROUND: Epidemics can be represented mathematically using a variety of models. One of these, the Kermack-McKendrick model, has been used to support health policy decisions concerning vaccination requirements. An unrelated body of literature suggests that some behaviours, including some types of violence, may spread in ways analogous to the contagious spread of infectious diseases, a process that has been characterized as "behavioural contagion". METHOD: Various parameter values reflecting the characteristics of crowds were substituted into the Kermack-McKendrick model. Computer simulations were used to evaluate the impact of these parameter values. RESULTS: The simulations reproduced several features of crowd violence: the tendency for riots to occur in large groups, the importance of rapid removal of violent individuals from crowds, and the roles of alcohol consumption and social identification processes. CONCLUSIONS: Epidemic models may be of relevance to the prevention and control of violent behaviour as they can assist with the identification of high-risk situations and prevention strategies. Theoretical constructs related to epidemic theory may have broad applicability for modelling the unstable course of some mental disorders. PMID- 15549237 TI - Epidemiology of multiple childhood traumatic events: child abuse, parental psychopathology, and other family-level stressors. AB - BACKGROUND: Multiple family-level childhood stressors are common and are correlated. It is unknown if clusters of commonly co-occurring stressors are identifiable. The study was designed to explore family-level stressor clustering in the general population, to estimate the prevalence of exposure classes, and to examine the correlation of sociodemographic characteristics with class prevalence. METHOD: Data were collected from an epidemiological sample and analyzed using latent class regression. RESULTS: A six-class solution was identified. Classes were characterized by low risk (prevalence=23%), universal high risk (7 %), family conflict (11 %), household substance problems (22 %), non nuclear family structure (24 %), parent's mental illness (13 %). CONCLUSIONS: Class prevalence varied with race and welfare status, not gender. Interventions for childhood stressors are person-focused; the analytic approach may uniquely inform resource allocation. PMID- 15549238 TI - Work and mental health. AB - BACKGROUND: Studies investigating the psychological correlates of types of occupation have focused on such disorders as stress, depression, suicide and substance abuse. There have also been some models proposed to allow understanding of factors common to different types of occupations. We sought to provide an overview of research related to work and mental health and consider future research directions. METHODS: A literature search was conducted using the Medline, PsycInfo, Embase and PubMed databases. The key words "occupation" or "work" were searched in combination with the key words "mental health", "risk factors", "disorders", "depression", "suicide", "trauma", "stress" or "substance use". RESULTS: Studies of "stress" tend to be more applicable to specific workplace issues. While some of the studies relating to onset of depression, suicide, substance abuse and trauma pertain to specific occupational issues and results are often not generalizable, they have progressed our understanding of risk factors to those disorders. There are workplace factors involving exposure to danger and crisis that lead to posttraumatic stress disorder (PTSD), substance abuse (including stimulants) and depersonalization. Workplace risk factors for depression involve situations promoting lack of autonomy, and involving "caring" for others as part of the work role, particularly where there is dependence on others for their livelihood. Risk factors for alcohol abuse include workplaces with access to alcohol and where use of alcohol is sanctioned. There appears to be a bi-directional relationship between personality and work, so that people are drawn to particular occupations, but the occupations then have an effect on them. An interactional model is proposed to consider this. CONCLUSION: The research questions pertaining to mental health are varied and will determine what mental health issues are of interest and the models of work applicable. There need to be more longitudinal studies and consideration of factors which the worker brings to the workplace (psychosocial issues, personality traits), as well as interpersonal issues and consideration of systemic, organizational, political and economic factors, including leadership styles. PMID- 15549239 TI - The contribution of epidemiology to the study of traumatic stress. AB - Post-traumatic stress disorder (PTSD) has been a controversial construct because of the complex set of factors that have been hypothesized to influence its onset and prevalence, such as compensation and withdrawal from combat duty. Epidemiology has done much to objectively clarify these controversies in the study of stratified population samples. The symptoms characterizing PTSD have been repeatedly described in large population samples where compensation is not a confounding issue and this has done much to support the validity of the construct. Epidemiology has also highlighted that the prevalence of exposure to traumatic events is far greater than was previously estimated. Emphasizing the importance of these events is accounting for the major burden of disease. Kessler (2000) has suggested the socio-economic effects of PTSD represent a burden of disease not dissimilar to that associated with depression. Traumatic events provide a unique opportunity to implement a preventative and public health approach to the management of psychiatric morbidity. Of particular importance is the apparent longevity of the influence that these events have on psychological adjustment. PMID- 15549240 TI - Psychosocial wellbeing and psychiatric care in the European Communities: analysis of macro indicators. AB - BACKGROUND: This paper reports the findings of an analytical comparison of several macro indicators collected routinely from institutional sources in Europe. METHODS: We carried out a review of macro indicators capable of providing a synthetic description of mental health status and the availability of psychiatric care in European countries. These were collected routinely from sources such as WHO, OECD, EUROSTAT, and IMS. The evaluation of temporal trends in each nation permits subsequent comparisons between countries. RESULTS: In all European countries, a decreased trend of suicides was observed in the period 1980 2000, with the exception of Ireland and (partially) of Spain. In Ireland, the increased trend was particularly strong, with a high risk in young and adult males. Portugal and Greece had respectively the highest and the lowest rates of undetermined causes of death. Most countries that were high consumers of alcohol in the initial period of observation had a rate above the national mean until 1980, after which consumption decreased. Most nations that were low consumers in 1960 had rates below the mean until 1980,when the rates progressively increased. However, a general decrease of alcohol-related deaths was apparent in all countries. Mortality associated with mental disorder increased, most clearly during the mid 1980s. The number of psychiatrists per 100,000 inhabitants ranged from 3.6 in Spain to 17.5 in Finland; of child psychiatrists, from 0.9 in Germany to 5.1 in Portugal. Psychiatric beds ranged from 0.4 per 1,000 inhabitants in Italy and Spain to 1.3 in Ireland. Nine countries showed a trend towards a reduction in psychiatric beds, whilst in a further five countries no change was observed. Rates of long-stay patients varied from 9.5 in the UK to 84 in Belgium, with a generally decreasing trend. The use of outpatient facilities differed markedly between countries, although there was a general increase over time. Official data were scarce in relation to the sale of psychotropic drugs. IMS data indicated increasing consumption in all European countries for antidepressant and antipsychotic medication. CONCLUSIONS: The official data resources available for European countries seem to indicate some discrepancies, probably relating to methods of recording. Better co-ordination in the collection of data about mental health status in the European Union and an improvement of the quality in available services is needed. PMID- 15549241 TI - Epidemiological theory, decision theory and mental health services research. AB - BACKGROUND: Mathematical models describing the epidemiology of major depression are potentially useful for epidemiological analyses, as decision support tools and in economic analyses. The objective of this project was to develop a Markov model based on epidemiological theory that may be useful for decision analysis and health services research. METHODS: Longitudinal data from a Canadian national survey, the National Population Health Survey (NPHS), were used. The NPHS has collected longitudinal data on a cohort of 17,262 subjects since 1994. The analysis employed a Markov tunnel in order to model the dependence of recovery probabilities on episode duration. RESULTS: Episode incidence ranged between 6.2 % per year in women under 35 to 0.26 % in men over the age of 65.A greater proportion of subjects over 35 years old reported episodes lasting more than 26 weeks. The probability of recovery declined with increasing episode duration, independently of sex. Under steady-state assumptions, a Markov model integrating these parameters predicted a point prevalence of approximately 2% in women and 1% in men under the age of 55. In older age groups, the predicted point prevalence declined in both sexes. CONCLUSIONS: These models support the hypothesis that sex differences in major depression prevalence are due primarily to differences in incidence rather than episode length. These results also indicate that there is no meaningful "central tendency" describing the distribution of episode length in major depression episode. Estimates of mean episode duration represent an intermixing of frequent brief episodes with infrequent protracted episodes. This finding may have important policy implications. PMID- 15549242 TI - Prevalence of psychiatric disorder in Europe: the potential and reality of meta analysis. AB - BACKGROUND: Within the European Mental Health Status Project, over 200 psychiatric surveys concerning members of the European Union (plus Norway) were examined for their potential for meta-analysis with regard to prevalence of psychiatric disorders and basic demographic and social variables. The diversity of samples, methods, analysis and presentation was such that only data derived from GHQ-12 and CIDI studies could be used, and those relating to sex differentials only. METHODS: The statistical program "Stata" was used to compute odds ratios (with confidence intervals) for individual studies, and to produce fixed and random effects estimates of the pooled odds ratio for all studies together, and a measure of heterogeneity. Forrest Plots were also produced. RESULTS: Analysis of GHQ-12 data with a cut-off point of 4, indicating a current or recent "probable mental health problem", showed, as expected, that women had higher prevalence rates than men. However, there was a relatively high heterogeneity score, suggesting that these studies may not be measuring the same thing. Analysis of CIDI results showed homogeneity for major depressive disorder within the last 12 months, with the risk for men about half of that for women. CONCLUSIONS: In terms of advancing epidemiological knowledge, the results are trivial, at most confirming what is already well known. However, the study shows the potential for pooled analysis, with much greater power in epidemiological investigation if consistency could be achieved in research. Various ways in which this might be done are discussed. It also shows the value of personal knowledge and personal networks in fields which are not well handled by electronic literature databases. PMID- 15549244 TI - The rebirth of PTSD: the rise of a new paradigm in psychiatry. AB - The new conception of psychological trauma that arose in the 1980s with the definition of PTSD in the DSM-III was a major change compared to the previous traumatic neurosis. While the clinical features were in some way similar, the political and sociological meanings of trauma were absolutely different. At that time, the invention of PTSD was much more the consequence of a broad mutation in mentality that introduced a new moral perspective in trauma studies than of a scientific discovery. In this paper, the author underlines from an anthropological point of view the second turning point that occurred in trauma studies in the mid 1990s when large epidemiological surveys did not confirm the first hypothesis. Readdressing the issues of vulnerability and risk factors that the previous version of PTSD had withdrawn, this second conception raises new epistemological questions that stay unsolved. PMID- 15549243 TI - Neuroticism: a non-informative marker of vulnerability to psychopathology. AB - BACKGROUND: Neuroticism measures are very popular in psychopathological research, but it is unclear how useful neuroticism is in studies of the aetiology of psychopathology. METHOD: A conceptual examination was made of the literature on the association of neuroticism and psychopathology, the ontological status of neuroticism, the purport of neuroticism questionnaires, and causal issues. RESULTS: The research on which neuroticism is built has historically been based solely on the factor analyses of the common adjectives used to describe usual behaviours. An abundance of studies have shown that neuroticism scores predict life stress, psychological distress, emotional disorders, psychotic symptoms, substance abuse, physical tension-related symptoms, medically unexplained physical symptoms, and health care utilisation. This evidence suggests that neuroticism scales index vulnerability to many forms of negative affect and psychiatric disorder. However, the associations do not clarify the nature of this vulnerability nor the underlying psychobiological mechanisms. We present evidence that neuroticism scores reflect a person's characteristic (or mean) level of distress over a protracted period of time. In this perspective, even prospective associations of neuroticism with mental health outcomes are basically futile, and largely tautological since scores on any characteristic with substantial within subject stability will predict, by definition, that characteristic and related variables at later points in time. CONCLUSION: Neuroticism is not an explanatory concept in the aetiology of psychopathology, since it measures a person's characteristic level of distress over a protracted period of time. This situation will not change until knowledge becomes available about: (i) the mechanisms that produce high neuroticism scores (and, therefore, also psychopathology) and (ii) its neurobiological substrate. Only then will we understand why neuroticism appears to "predict" the outcomes it predicts. PMID- 15549245 TI - Childhood sexual abuse predicts poor outcome seven years after parasuicide. AB - BACKGROUND: There is substantial empirical research linking borderline personality disorder with prolonged mental instability and recurrent suicidality. At the same time, a growing body of observations links borderline personality disorder to sexual abuse and other forms of abuse and trauma in childhood. The aim of this study was to investigate among patients admitted for parasuicide the predictive value for outcome 7 years after the parasuicide of a diagnosis of borderline personality disorder compared to the predictive value of a history of childhood sexual abuse. METHODS: Semi-structured interviews were conducted at the time of the index parasuicide, with follow-up interviews 7 years later. In addition, information was collected from medical records at the psychiatric clinic. A logistic regression analysis was used to assess the specific influence of the covariates borderline personality disorder, gender and reported childhood sexual abuse on the outcome variables. RESULTS: Univariate regression analysis showed higher odds ratios for borderline personality disorder, female gender and childhood sexual abuse regarding prolonged psychiatric contact and repeated parasuicides. A combined logistic regression model found significantly higher odds ratios only for childhood sexual abuse with regard to suicidal ideation, repeated parasuicidal acts and more extensive psychiatric support. CONCLUSION: The findings support the growing body of evidence linking the characteristic symptoms of borderline personality disorder to childhood sexual abuse, and identify sexual abuse rather than a diagnosis of borderline personality disorder as a predictor for poor outcome after a parasuicide. The findings are relevant to our understanding and treatment of parasuicide patients, especially those who fulfil the present criteria for borderline personality disorder. PMID- 15549246 TI - Double depression in an Australian population. AB - BACKGROUND: Double depression, or dysthymia with superimposed major depression, is a major public health issue that imposes considerable burden on the community. Double depression and its associated morbidity have not previously been delineated in an Australian population. METHODS: A random and representative sample of the South Australian population was assessed by trained interviewers. The mood module of the Primary Care Evaluation of Mental Disorders (PRIME-MD), the Short-Form Health Status Questionnaire (SF-36), and Assessment of Quality of Life (AQoL) instruments were administered, and data related to treatment use and role functioning were collated. RESULTS: Double depression was present in 2.2% of the population. This group reported high levels of treatment-seeking behaviour with 90% seeking treatment in the last month and 42.4 % taking antidepressants. They also had a highly significantly poorer quality of life than did others in the community. CONCLUSIONS: The 2.2% of the population with double depression reported high use of services with poor functioning and health-related quality of life. More effective intervention strategies are required. PMID- 15549247 TI - Risk of non-fatal suicide ideation and behaviour in recent onset schizophrenia- the influence of clinical, social, self-esteem and demographic factors. AB - BACKGROUND: Suicide rates amongst schizophrenic patients are high. There are disadvantages in investigating successfully completed suicides which make suicidal ideation and previous attempts important proxy measures of suicidal risk. The aim of this study was to investigate factors associated with these risk measures. METHOD: Fifty-nine patients suffering recent onset schizophrenia were assessed for suicidal ideation and history, and a range of demographic, clinical, social (including relatives' Expressed Emotion) and self-esteem measures. Univariate comparisons were made between those with and without suicide ideation and previous attempts. Path analysis was conducted to identify factors directly or indirectly associated with a composite scale of risk (low, medium or high). RESULTS: Approximately 25% of the sample reported a current desire to kill themselves and 47% had made one or more previous attempts. There were numerous significant univariate differences between those with or without ideation or history. Path analysis indicated that greater hopelessness (OR 1.22) and longer duration of illness (OR 1.13) increased risk. Hopelessness was associated with higher negative self-evaluation and social isolation. Negative self-evaluation was associated with more relatives' criticism which was associated with more negative symptoms. Being a male, unmarried and unemployed were all significantly associated with an increase in negative symptoms. Social isolation was associated with being unemployed, older, more positive symptoms and longer illness duration. Duration of illness was not itself predicted by any other variables. CONCLUSION: Non-fatal suicide ideation and behaviour are significantly associated with an array of demographic, clinical, interpersonal and psychological factors. To reduce risk of suicide, these factors need to be assessed and methods developed to reduce their influence. PMID- 15549248 TI - [Hip resurfacing arthroplasty]. AB - In comparison to stemmed total hip replacements, hip resurfacing offers advantages especially in joint stability and amount of femoral bone resection. After the poor results achieved with this concept that were mainly caused by failure of the materials used, reintroduction of the metal-on-metal bearing initiated a renaissance. This bearing, the cementless cup, and the improved surgical technique led to better short- to medium-term results. Revision and complication rates are now comparable to conventional total hip replacements. The functional capacity of the method is higher. Because long-term results are not available, however, questions remain, for instance, the consequences of the higher metal ion serum concentrations or the impossibility of changing the inlay when femoral revision becomes necessary. PMID- 15549249 TI - [Early functional results with the Mayo-hip, a short stem system with metaphyseal intertrochanteric fixation]. AB - Total hip arthroplasty has become one of the most successful standard procedures in orthopedic surgery. With a more frequent use in young and active patients bone saving procedures become more important. The goal is to save good bone stock for the revision procedure. One example of conservative femoral implants is the Mayo stem with reported long term results. The stem design allows a metaphyseal intertrochanteric multipoint fixation for primary fixation. The surgical technique is simple. Using a transgluteal approach, the medial preparation of the femoral neck decreases significantly the irritation of the abductor tendons. In a prospective-randomized study, the early functional results with the Mayo-stem were significantly better than the results achieved with a cement-free standard stem. The Mayo-stem may not be indicated for every hip situation. Its use, however, in cases with normal anatomy contributes to save autologeous bone. Therefore, this short stem is a sensible alternative to standard stems. PMID- 15549250 TI - [Telomerase activity and telomere length of human mesenchymal stem cells. Changes during osteogenic differentiation]. AB - Human mesenchymal stem cells (hMSC) exhibit properties of self-renewal and differentiation. Assuming that telomerase activity is associated with self renewal, it might be useful to identify and define hMSC on the basis of their telomerase status. However, telomerase activity in hMSC remains a controversial issue. Therefore, the aim of our study was to investigate telomerase activity in proliferating and highly proliferating hMSC and to measure telomerase activity and changes in telomere restriction fragment (TRF) length of confluent hMSC and of osteogenically differentiated hMSC. For tissue engineering applications scaffolds should be seeded with cells that have not lost their ability to self replicate and differentiate during in vitro cell culture. Telomerase activity could be used to characterise and isolate these cells. PMID- 15549251 TI - [In vivo and in vitro bone regeneration from cord blood derived mesenchymal stem cells]. AB - BACKGROUND: Mesenchymal stem cells with an osteoblastic differentiating potency are investigated in regard of probable tissue engineering for further clinical application. The following report describes the use of cord blood derived stem cells as an alternative to other stem cell populations for bone regenerating tissue engineering. METHODS: To demonstrate the multipotency of cord blood derived mesenchymal stem cells, unrestringated somatic stem cells (USSC) were isolated from cord blood and underwent an osteo-, chondro- and adipoblastic in vitro stimulation. To evaluate the osteoinductive potency of a porcine collagen I/III cell carrier USSC were incubated on this matrix. To investigate the in vivo effects of human USSC an athymic rat model was developed. These cells were transplanted into a femoral defect. RESULTS: Cord blood derived mesenchymal stem cells (USSC) have an in vitro multipotency and show adipo-, chondro- and osteogenic differentiation. The porcine collagen I/III carrier promoted an osteoblastic differentiation. USSC survived after xenotransplantation in an athymic rat and differentiated into osteoblasts filling the bony defect zone. CONCLUSION: Human USSC are a mesenchymal multipotent stem cell population that shows osteoblastic differentiation onto a collagen I/III carrier in vitro as well as in an athymic rat in vivo. PMID- 15549254 TI - Consensus meeting on microdialysis in neurointensive care. AB - BACKGROUND: Microdialysis is used in many European neurointensive care units to monitor brain chemistry in patients suffering subarachnoid hemorrhage (SAH) or traumatic brain injury (TBI). DISCUSSION: We present a consensus agreement achieved at a meeting in Stockholm by a group of experienced users of microdialysis in neurointensive care, defining the use of microdialysis, placement of catheters, unreliable values, chemical markers, and clinical use in SAH and in TBI. CONCLUSIONS: As microdialysis is maturing into a clinically useful technique for early detection of cerebral ischemia and secondary brain damage, there is a need to following such definition regarding when and how to use microdialysis after SAH and TBI. PMID- 15549252 TI - Clinical identification of cognitive impairment in ICU survivors: insights for intensivists. AB - BACKGROUND: A growing body of research has demonstrated the presence of ongoing cognitive impairment in large numbers of ICU survivors. OBJECTIVE: This review offers a practical framework for practicing intensivists and those following patients after their ICU stay for the identification of cognitive impairment in ICU survivors. CONCLUSIONS: Early detection of cognitive impairment in critically ill patients is an important and achievable goal, but overt cognitive impairment remains unrecognized in most cases. However, it can be identified by objective (test scores) or subjective evidence (clinical judgment, patient observation, family interaction). PMID- 15549256 TI - Severe cartilage damage by broken poly-L-lactic acid (PLLA) interference screw after ACL reconstruction. AB - Poly-L-lactic acid (PLLA) bioabsorbable interference screws are widely used for fixation of tendon to bone and bone to bone in anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) reconstructions. Complications are rare. To our knowledge this is the first report of severe chondral damage caused by late breakage of the screw. Breakage of bioscrews has only been published in cases with tendon to bone fixation. PMID- 15549257 TI - Functional outcomes of primary and secondary repairs of vesicovaginal fistulae via vaginal cuff scar excision. AB - Hospital and office charts of patients who underwent vaginal cuff scar excision for vesicovaginal fistula (VVF) repair from February 1998 to December 2002 at our institution were reviewed. Preoperative demographics and fistula characteristics were gathered. Intraoperative data included use of tissue flaps, blood loss, OR time and anesthetic type. Postoperative review included time to discharge, successful repair and postoperative urinary or sexual dysfunction. Forty fistula repairs were identified. Ninety-three percent occurred after a hysterectomy and no subjects had a history of radiation. Forty-two percent had failed at least one surgical repair of their fistula and 12% had failed two or more attempted repairs. Twenty percent of the fistulae measured 1 cm or more in diameter and the remaining 80% were 5 mm or less. Peritoneal flaps and martius flaps were performed in 32% and 5%, respectively. Postoperatively, 100% of subjects were evaluated at 3 weeks when the suprapubic catheter was removed and 93% were evaluated at 3 months or later. All subjects were cured of their fistulae at last contact. At 3 months postoperatively, 94% percent denied any urinary dysfunction and 85% had resumed sexual intercourse. Two sexually active subjects reported mild deep dyspareunia. Transvaginal cuff scar excision is an effective method for the primary and secondary repair of vesicovaginal fistulae and does not appear to cause postoperative irritative voiding symptoms or dyspareunia. PMID- 15549258 TI - Factors that affect recurrence after anterior colporrhaphy procedure reinforced with four-corner anchored polypropylene mesh. AB - The purpose of this study was to evaluate the effectiveness of the anterior colporrhaphy procedure reinforced with four-corner anchored polypropylene mesh in patients with severe (stage III or IV) anterior vaginal prolapse. Thirty-eight consecutive women were enlisted for this prospective study. The procedure consisted of an extensive vaginal dissection to join the vesicovaginal and retropubic space and an anchoring of a polypropylene mesh patch between the two Arcus Tendineus Fasciae Pelvis in a tension-free manner. The mean age of the study group was 63 (33-80) years. The success rate was 87% (33/38) at a mean follow-up interval of 21 (12-29) months. A total of eight (100%) patients were also cured of concomitant stress incontinence (five overt and three occult type) with an additional tension-free vaginal tape (TVT) operation. During follow-up, there were five de-novo stress incontinence cases (16.7%) and four vaginal erosions of mesh (10.5%). Four clinical variables--diabetes mellitus, recurrent anterior vaginal prolapse, chronic cough and vaginal erosions of mesh--were found to have a significant correlation with an unsatisfactory surgical result with large values of hazard ratios found by survival analysis. We concluded that the anterior colporrhaphy procedure reinforced with four-corner anchored polypropylene mesh was effective for most, but failed in some patients who had specific risk factors within short convalescence periods. Concomitant stress incontinence can be successfully treated by a TVT operation in combination with the anterior colporrhaphy procedure reinforced with four-corner anchored polypropylene mesh. However, the anterior colporrhaphy procedure may itself have adverse effects on urethral sphincter function. PMID- 15549259 TI - Surgical treatment of ureteral obstruction from endometriosis: our experience with thirteen cases. AB - Endometriosis is a biologically benign albeit aggressive pathology marked by high local recurrences. Ureteral involvement accounts for only a minority of cases (0.1-0.4%) with often non-specific symptoms at clinical presentation and difficult preoperative diagnosis. Thirteen cases of severe ureteral endometriosis (i.e. causing significant obstruction to the urinary flow) were observed and surgically treated, out of 17 ureteral units affected (three cases of bilateral involvement, one case of complete pyeloureteral duplicity). The initial symptomatology was acute and related to ureteral obstruction in eight cases, silent and non-specific in the other five; a presumptive diagnosis was made only for the seven patients (53.9%) with a positive medical history for pelvic (and in two cases also ureteral) endometriosis. Preoperative drainage of urine proved necessary for eight patients due to the complete functional exclusion of the excretory axis. One patient (7.7%) underwent nephrectomy due to renal atrophy. Segmental ureteral resection and termino-terminal anastomosis were performed in two patients, while seven patients underwent segmental ureterectomy and ureterocystoneostomy, with bladder psoas hitching in four cases and vesical flap according to Casati-Boari in one case. All three cases of bilateral involvement were treated by bilateral segmental ureterectomy and trans-uretero-uretero cystoneostomy with bladder psoas hitching. Following histological examination, all patients were diagnosed with active ureteral endometriosis, which was found to be intrinsic in five cases (38.5%) and extrinsic in the other eight. One of the two patients that had undergone ureterectomy and termino-terminal anastomosis had to undergo ureteral resection and ureterocystoneostomy 22 months later due to relapsing endometriosis-induced stenosis. Conversely, no ureteral endometriosis relapses occurred in the remaining 12 patients within the mean follow-up time of 41.1 months (range 6-91). Ureteral endometriosis is marked by non-specific symptoms, making preoperative diagnosis often difficult. Therefore, an ultrasound or urographic examination of the urinary tract in case of pelvic endometriosis is absolutely essential. In our experience, terminal ureterectomy with ureterocystoneostomy has provided long-term favourable results as extended ureteral resection can be performed and continuity of the urinary tract can be restored without resorting to the distal pelvic ureter, which is often affected by the disease besides being more subject to relapses. PMID- 15549260 TI - Preoperative maximal flow rate may be a predictive factor for the outcome of tension-free vaginal tape procedure for stress urinary incontinence. AB - This study examined factors potentially predicting on the outcome of the tension free vaginal tape procedure for stress urinary incontinence. A total of 60 women (aged 35-71 years, mean 57) with at least follow-up of 2 years were included in the study. The tension-free vaginal tape procedure was performed under local anesthesia except in two patients. At the latest follow-up 50 (83.3%) were cured of stress urinary incontinence. Two patients had new-onset urge symptoms without urge incontinence episodes. Multivariate regression analysis showed maximal flow rate to be associated with 0.9-fold risk of the failure; no other parameters had statistical significance. Our results confirm the feasibility and safety of the tension-free vaginal tape procedure and demonstrate that preoperative low maximal flow rate may be associated with the failure of the tension-free vaginal tape procedure. PMID- 15549261 TI - Impact of radical hysterectomy for cervical cancer on urodynamic findings. AB - To elucidate the impact of radical hysterectomy upon the urodynamic findings of patients with cervical cancer, 20 patients with cervical cancer at stage IB to IIA who underwent radical hysterectomy were recruited. Each patient underwent a 20-min pad test and urodynamic study prior to and 3 months after radical hysterectomy. ANOVA, Bonferroni test and paired t -test were utilized for analysis. The mean age of the 20 patients was 50.2+/-8.7 years with a mean parity of 3.5+/-1.5. Four (20%) of the 20 cases revealed normal urodynamic findings preoperatively, and the urodynamic findings became abnormal after surgery. Comparing the urodynamic parameters of both bladder voiding and storage functions pre- and post-surgery, we found significant impairments postoperatively in all 20 cases. Our data demonstrate that abnormal urodynamic findings may pre-exist for some patients with cervical cancer prior to surgical treatment. These findings may worsen, and/or additional abnormal states may arise subsequent to radical hysterectomy. PMID- 15549262 TI - Does posture affect cystometric parameters and diagnoses? AB - The objective of this study was to investigate the effect of lying and sitting positions on urodynamic parameters and diagnoses. This prospective study was carried out on 96 women with urinary incontinence who underwent urodynamic assessment. Cystometry was performed both in the lying and sitting positions. For filling cystometry, we infused normal saline at a rate of 50 ml/min. All the results were entered on the urodynamic database and were analysed using Minitab software release 13.30. Mean age was 49 (20-84) years. Sixty-four (67%) women complained of mixed incontinence, 16 (17%) of urgency alone, eight (8%) of stress incontinence and eight (8%) of urgency and urge incontinence. Two (2%) showed stress incontinence by lying cystometry, and 53 (55%) by sitting cystometry. During lying nine (9%) demonstrated detrusor overactivity, while 53 (55%) demonstrated detrusor overactivity in sitting position. No case of mixed incontinence was diagnosed by lying cystometry but 17 (18%) cases were detected by sitting cystometry. This study explains the higher detection rate of stress incontinence, detrusor overactivity and mixed incontinence by cystometry in sitting position. Therefore, we recommend that sitting posture is preferred over lying position for performing cystometry. PMID- 15549263 TI - mRNA assessment for procollagen production in women with genuine stress urinary incontinence. AB - The aim of the study was to examine changes in the levels of mRNA for procollagen type I and III in women with or without genuine stress incontinence (GSI). The study was performed in the 2nd Department of Obstetrics and Gynecology of University of Athens. Sixty-eight women participated in the study and they were divided in two groups. We did not find statistically significant difference between the two groups of patients in relation to the amount of mRNA of procollagen type I and III (p>0.05), but the quantity of collagen type I and III was significantly reduced in patients with stress incontinence (p<0.05). The possible cause for the reduction in the amount of collagen in women with GSI could be attributed to either a disturbance in the translation of mRNA to protein (collagen) or increased catabolism of collagen by its collagenase. PMID- 15549264 TI - Conservative management of vesicouterine fistula after uterine rupture. AB - We report the closure of a vesicouterine fistula with conservative management utilizing an indwelling transurethral Foley catheter. Uterine rupture occurred during a trial of vaginal birth after cesarean section, necessitating an emergency cesarean section. Upon entry into the abdomen, the base of the bladder was noted to be involved in the uterine rupture. The bladder trigone and ureteral orifices appeared normal. A primary, two-layer bladder repair was performed. A cystogram on postoperative day 14 demonstrated a vesicouterine fistula. Conservative management involving bladder drainage for 21 days with a transurethral Foley catheter was successful in closure of the fistula. Vesicouterine fistula, a documented complication of uterine rupture due to attempted vaginal birth after previous cesarean section, can spontaneously resolve with conservative management alone. PMID- 15549265 TI - Recurrent pelvic organ prolapse in a woman with bladder exstrophy: a case report of surgical management and review of the literature. AB - Management of gynecologic issues in women with bladder exstrophy is challenging. Congenital pelvic organ prolapse and prolapse-associated pregnancy and delivery are common. The management of prolapse is complicated by the anatomic changes involving the bony pelvis, connective tissue support, pelvic floor, and the length and axis of the vagina. A case of recurrent pelvic organ prolapse successfully managed with a sacral colpopexy and a review of the literature are presented. PMID- 15549266 TI - Balance disorder and increased risk of falls in osteoporosis and kyphosis: significance of kyphotic posture and muscle strength. AB - This controlled trial was designed to investigate the influence of osteoporosis related kyphosis (O-K) on falls. Twelve community-dwelling women with O-K (Cobb angle, 50-65 degrees measured from spine radiographs) and 13 healthy women serving as controls were enrolled. Mean age of the O-K group was 76 years (+/ 5.1), height 158 cm (+/-5), and weight 61 kg (+/-7.9), and mean age of the control group was 71 years (+/-4.6), height 161 cm (+/-3.8), and weight 66 kg (+/ 11.7). Quantitative isometric strength data were collected. Gait was monitored during unobstructed level walking and during stepping over an obstacle of four different heights randomly assigned (2.5%, 5%, 10%, and 15% of the subject's height). Balance was objectively assessed with computerized dynamic posturography consisting of the sensory organization test. Back extensor strength, grip strength, and all lower extremity muscle groups were significantly weaker in the O-K group than the control group (P <0.05), except right ankle plantar flexors (P =0.09). There was a significant difference in the anteroposterior and mediolateral displacements and velocities. The O-K subjects had less anteroposterior displacement, greater mediolateral displacement, reduced anteroposterior velocity, and increased mediolateral velocity compared with controls for all conditions of unobstructed and obstructed level walking. Obstacle height had a significant effect on all center-of-mass variables. The O-K subjects had significantly greater balance abnormalities on computerized dynamic posturography than the control group (P =0.002). Data show that thoracic hyperkyphosis on a background of reduced muscle strength plays an important role in increasing body sway, gait unsteadiness, and risk of falls in osteoporosis. PMID- 15549267 TI - Chlorosis during nitrogen starvation is altered by carbon dioxide and temperature status and is mediated by the ClpP1 protease in Synechococcus elongatus. AB - The interactive effects of inorganic carbon status, temperature and light on chlorosis induced by nitrogen deficiency, and the roles of Clp proteases in this process were investigated. In wild-type cultures grown in high or ambient CO(2), following transfer to media lacking combined nitrogen, phycocyanin per cell dropped primarily through dilution of the pigment through cell division, and also suffered variable degrees of net degradation. When grown at high CO(2) (5%), chlorophyll (Chl) suffered net degradation to a greater extent than phycocyanin. In marked contrast, growth at ambient CO(2) resulted in Chl per cell dropping through dilution. Conditions that drove net Chl degradation in the wild-type resulted in little or no net Chl degradation in a clpPI inactivation mutant, with Chl content dropping largely through growth dilution in the mutant. The chlorotic response of a clpPII inactivation strain was nearly the same as that of wild type, although phycocyanin degradation may have been slightly accelerated in the former. PMID- 15549268 TI - Phylogeny of symbiotic cyanobacteria within the genus Nostoc based on 16S rDNA sequence analyses. AB - A phylogenetic analysis of selected symbiotic Nostoc strain sequences and available database 16S rDNA sequences of both symbiotic and free-living cyanobacteria was carried out using maximum likelihood and Bayesian inference techniques. Most of the symbiotic strains fell into well separated clades. One clade consisted of a mixture of symbiotic and free-living isolates. This clade includes Nostoc sp. strain PCC 73102, the reference strain proposed for Nostoc punctiforme. A separate symbiotic clade with isolates exclusively from Gunnera species was also obtained, suggesting that not all symbiotic Nostoc species can be assigned to N. punctiforme. Moreover, isolates from Azolla filiculoides and one from Gunnera dentata were well nested within a clade comprising most of the Anabaena sequences. This result supports the affiliation of the Azolla isolates with the genus Anabaena and shows that strains within this genus can form symbioses with additional hosts. Furthermore, these symbiotic strains produced hormogonia, thereby verifying that hormogonia formation is not absent in Anabaena and cannot be used as a criterion to distinguish it from Nostoc. PMID- 15549269 TI - FieF (YiiP) from Escherichia coli mediates decreased cellular accumulation of iron and relieves iron stress. AB - The Escherichia coli yiiP gene encodes an iron transporter, ferrous iron efflux (FieF), which belongs to the cation diffusion facilitator family (CDF). Transcription of fieF correlated with iron concentration; however, expression appeared to be independent of the ferrous iron uptake regulator Fur. Absence of FieF led to decreased growth of E. coli cells in complex growth medium but only if fur was additionally deleted. The presence of EDTA was partially able to relieve this growth deficiency. Expression of fieF in trans rendered the double deletion strain more tolerant to iron. Furthermore, E. coli cells exhibited reduced accumulation of (55)Fe when FieF was expressed in trans. FieF catalyzed active efflux of Zn(II) in antiport with protons energized by NADH via the transmembrane pH gradient in everted membrane vesicles. Using the iron-sensitive fluorescent indicator PhenGreen-SK encapsulated in proteoliposomes, transmembrane fluxes of iron cations were measured with purified and reconstituted FieF by fluorescence quenching. This suggests that FieF is an iron and zinc efflux system, which would be the first example of iron detoxification by efflux in any organism. PMID- 15549270 TI - Quinapril effects on resistance artery structure and function in hypertension. AB - The effects of chronic treatment with quinapril on blood pressure, vascular reactivity and structure of resistance arteries were examined in adult, male spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) rats. SHR and WKY at 15 weeks of age were treated with quinapril (10 mg/kg per day) for 10 weeks. Structural changes in the mesenteric arteries were measured by optical sectioning with confocal microscopy and in renal arteries by light microscopic measurements. Apoptotic cells in the mesenteric vessel wall were identified using the terminal deoxynucleotide transferase-mediated dUTP-nick end-labelling (TUNEL) method. The response of mesenteric arteries to noradrenaline, electrical stimulation, acetylcholine and sodium nitroprusside was studied using a pressure myograph system. Treatment with quinapril significantly lowered systolic blood pressure and ventricular weight in both SHR and WKY. It reduced wall thickness and medial volume in mesenteric arteries from SHR and WKY and media-to-lumen ratio in interlobular arteries of SHR. It also decreased the number of smooth muscle layers in SHR and increased the number of apoptotic smooth muscle cells in both SHR and WKY. In addition, treatment normalized the augmented contractile responses and improved the impaired relaxation response of SHR mesenteric arteries to the level of WKY. We conclude that treatment with quinapril lowered blood pressure and improved cardiac and vessel structure and vessel function. An increase in apoptotic process of medial smooth muscle cells is one of the mechanisms underlying the vascular structural improvement. PMID- 15549271 TI - Selective sensory denervation by capsaicin aggravates adriamycin-induced cardiomyopathy in rats. AB - Capsaicin-sensitive sensory nerves that contain calcitonin gene-related peptide (CGRP) contribute significantly to cardioprotective mechanisms. In this study, the possible role of capsaicin-sensitive afferent nerves in the development of congestive heart failure was examined in an established model of adriamycin induced experimental cardiomyopathy in rats. Systemic treatment with capsaicin was utilized to deplete sensory neuropeptides from cardiac afferent nerves. Echocardiography was applied to assess the cardiac function in adriamycin-treated rats pretreated with capsaicin or its vehicle. In control rats, adriamycin treatment produced a reduction in the fractional shortening of the left ventricle and an increase in the ratio of the left atrial diameter and the aortic diameter, indicative of a decreased myocardial contractility and heart failure only at 3-4 weeks post-treatment. In contrast, in capsaicin-pretreated rats, a deterioration of the cardiac function was already evident 1 week after the cessation of adriamycin administration, while the clinical signs associated with cardiomyopathy were more severe and displayed a significantly more rapid progression. Immunohistochemistry revealed a complete depletion of calcitonin gene-related peptide from cardiac sensory nerves after systemic capsaicin treatment. This study has demonstrated that elimination of capsaicin-sensitive afferent nerves promotes the development and progression of adriamycin-induced myocardial dysfunction. The results suggest that interfering with capsaicin/vanilloid receptor function and/or perturbation of the myocardial CGRP metabolism may open up new perspectives concerning prevention and/or alleviation of the pathological changes that follow adriamycin treatment. PMID- 15549272 TI - Inhibition of TRPM2 channels by the antifungal agents clotrimazole and econazole. AB - TRPM2 is a Ca(2+)-permeable non-selective cation channel that uniquely is activated by intracellular ADP-ribose. To date, only one pharmacological blocker of this channel, namely flufenamic acid (FFA), has been described. Here we demonstrate, using patch clamp electrophysiology, that the antifungal imidazoles clotrimazole and econazole inhibit ADP-ribose-activated currents in HEK-293 cells expressing recombinant human TRPM2 (hTRPM2). For both compounds, all concentrations in a range from 3 microM to 30 microM produced an essentially complete inhibition of the TRPM2-mediated current. The rate of current antagonism was dependent on the concentration applied, with higher concentrations producing faster block. In addition, decreasing extracellular pH accelerated inhibition of TRPM2 by both clotrimazole and econazole; extracellular alkalisation produced the converse effect. Additional experiments indicated hTRPM2 activation was required for the antagonism of either compound to develop, and that neither compound blocked from the intracellular face of the plasma membrane. ADP-ribose-activated whole-cell and single-channel currents in the rat insulinoma cell-line CRI-G1 were also antagonised by clotrimazole. Contrary to the observations made with hTRPM2, antagonism in CRI-G1 cells could be largely reversed following clotrimazole removal. These experiments suggest that imidazole antifungals may be useful tool antagonists for future studies of TRPM2 function. PMID- 15549273 TI - Urotensin-II-mediated cardiomyocyte hypertrophy: effect of receptor antagonism and role of inflammatory mediators. AB - Urotensin-II (U-II), the most potent mammalian vasoconstrictor identified, and its receptor, UT, exhibits increased expression in cardiac tissue and plasma in congestive heart failure (CHF) patients. Cardiomyocyte hypertrophy is primarily responsible for increased myocardial mass associated with cardiac injury. Neurohumoral factors such as angiotensin-II, endothelin-1, catecholamines, and inflammatory cytokines are thought to mediate this response. U-II shares similar biological activities with other hypertrophic G(q)-coupled receptor ligands such as angiotensin-II and endothelin-1, but a role for U-II in cardiomyocyte hypertrophy has not been characterized. The hypothesis of the current study was that U-II, acting through its G(q)-coupled receptor UT plays a hypertrophic role in cardiac hypertrophic remodeling. We report that adenoviral upregulation of the UT receptor "unmasked" U-II-induced hypertrophy in H9c2 cardiomyocytes, with a threshold response of 202+/-8 binding sites/cell. U-II was equally as efficacious as phenylephrine in inducing hypertrophy, measured by a reporter assay (EC(50) 0.7+/-0.2 nM) and [(3)H]-leucine incorporation (EC(50) 150+/-40 nM). A competitive peptidic UT receptor antagonist, BIM-23127, inhibited U-II-induced hypertrophy ( K(B) 34+/-6 nM). U-II did not affect cell proliferation or apoptosis, indicating that U-II is more hypertrophic than apoptotic or hyperplastic in cardiomyocytes. U-II (10 nM) stimulated interleukin-6 release in UT-expressing cardiomyocytes (4.6-fold at 6 h). Finally, in a rat heart failure model, cardiac ventricular mRNA expression of U-II, UT receptor, interleukin-6, and interleukin-1-beta is increased time-dependently following myocardial injury. These results indicate that U-II might play a role in cardiac remodeling associated with CHF by stimulation of cardiomyocyte hypertrophy via UT, and through upregulation of inflammatory cytokines. As such, UT antagonism may represent a novel therapeutic target for the clinical management of heart failure. PMID- 15549274 TI - Kidney function in mice: thiobutabarbital versus alpha-chloralose anesthesia. AB - Mice that lack or over-express a gene of interest are important tools for unraveling gene function. The determination of single nephron function by micropuncture or precise determination of glomerular filtration rate (GFR) by inulin clearance method require experiments under anesthesia. A good anesthetic protocol should allow for reasonable and stable glomerular and tubular function. The aim of this study was to compare the commonly used thiobutabarbital (TBB) versus alpha-chloralose (CHL) anesthesia with regard to absolute levels and the stability of blood pressure, heart rate, and kidney function. Male CD1 mice were anesthetized with TBB (100 mg/kg body weight i.p.) or CHL (120 mg/kg body weight i.p.), plus ketamine (100 mg/kg body weight i.m.) given to every mouse for analgesia. After preparation for clearance experiments, two 30-min urine collections were performed at periods 1 and 2 (P1 and P2). It was observed that heart rate and mean arterial blood pressure did not differ between TBB ( n=9) vs. CHL ( n=9) and were stable through P1 and P2. In CHL, GFR as well as fractional excretion of fluid, Na(+) and K(+) were stable from P1 to P2 (P1: 190+/-15 microl/min, 1.6+/-0.2%, 0.7+/-0.1%, 35+/-5%; percent change in P2: 1+/-6, 26+/ 10, 29+/-15, 6+/-10 respectively). In TBB, GFR was significantly greater vs. CHL in P1 and did not significantly change in P2 (246+/-8 microl/min, p<0.05; percent change: -6.5+/-4). Fractional excretion of fluid, Na(+) and K(+) were not significantly different vs. CHL in P1, but significantly increased in P2 (P1: 1.5+/-0.2%, 1.1+/-0.2%, 31+/-3%; percent change in P2: 122+/-23, 128+/-21 and 29+/-6 respectively; each p<0.05 vs. P1). In conclusion, mice under both anesthetic regimens present reasonable and stable blood pressure and reasonable kidney function, but kidney reabsorption is more stable under CHL than under TBB anesthesia, which may facilitate study of the response in kidney function to acute interventions. PMID- 15549275 TI - Cognitive and physiological effects of an "energy drink": an evaluation of the whole drink and of glucose, caffeine and herbal flavouring fractions. AB - RATIONALE: Both glucose and caffeine can improve aspects of cognitive performance and, in the case of caffeine, mood. There are few studies investigating the effects of the two substances in combination. OBJECTIVES: We assessed the mood, cognitive and physiological effects of a soft drink containing caffeine and glucose as well as flavouring levels of herbal extracts. The effects of different drink fractions were also evaluated. METHODS: Using a randomised, double-blind, balanced, five-way crossover design, 20 participants who were overnight fasted and caffeine-deprived received 250 ml drinks containing 37.5 g glucose; 75 mg caffeine; ginseng and ginkgo biloba at flavouring levels; a whole drink (containing all these substances) or a placebo (vehicle). Participants were assessed in each drink condition, separated by a 7-day wash-out period. Cognitive, psychomotor and mood assessment took place immediately prior to the drink then 30 min thereafter. The primary outcome measures included five aspects of cognitive performance from the Cognitive Drug Research assessment battery. Mood, heart rate and blood glucose levels were also monitored. RESULTS: Compared with placebo, the whole drink resulted in significantly improved performance on "secondary memory" and "speed of attention" factors. There were no other cognitive or mood effects. CONCLUSIONS: This pattern of results would not be predicted from the effects of glucose and caffeine in isolation, either as seen here or from the literature addressing the effects of the substances in isolation. These data suggest that there is some degree of synergy between the cognition-modulating effects of glucose and caffeine which merits further investigation. PMID- 15549276 TI - Methylxanthines are the psycho-pharmacologically active constituents of chocolate. AB - RATIONALE: Liking, cravings and addiction for chocolate ("chocoholism") are often explained through the presence of pharmacologically active compounds. However, mere "presence" does not guarantee psycho-activity. OBJECTIVES: Two double-blind, placebo-controlled studies measured the effects on cognitive performance and mood of the amounts of cocoa powder and methylxanthines found in a 50 g bar of dark chocolate. METHODS: In study 1, participants ( n=20) completed a test battery once before and twice after treatment administration. Treatments included 11.6 g cocoa powder and a caffeine and theobromine combination (19 and 250 mg, respectively). Study 2 ( n=22) comprised three post-treatment test batteries and investigated the effects of "milk" and "dark" chocolate levels of these methylxanthines. The test battery consisted of a long duration simple reaction time task, a rapid visual information processing task, and a mood questionnaire. RESULTS: Identical improvements on the mood construct "energetic arousal" and cognitive function were found for cocoa powder and the caffeine+theobromine combination versus placebo. In chocolate, both "milk chocolate" and "dark chocolate" methylxanthine doses improved cognitive function compared with "white chocolate". The effects of white chocolate did not differ significantly from those of water. CONCLUSIONS: A normal portion of chocolate exhibits psychopharmacological activity. The identical profile of effects exerted by cocoa powder and its methylxanthine constituents shows this activity to be confined to the combination of caffeine and theobromine. Methylxanthines may contribute to the popularity of chocolate; however, other attributes are probably much more important in determining chocolate's special appeal and in explaining related self-reports of chocolate cravings and "chocoholism". PMID- 15549277 TI - Stimulant and relaxant drugs combined with stimulant and relaxant information: a study of active placebo. AB - RATIONALE: The active placebo hypothesis states that placebo effects are potentiated when an active drug is administered. OBJECTIVE: This hypothesis was tested in an experiment where information about the effect of a drug was combined with administration of an active drug or placebo. METHODS: Information that a drug acted as a relaxant, a stimulant, or as a placebo was crossed with oral administration of a relaxant drug (700 mg carisoprodol), a stimulant drug (400 mg caffeine) or placebo (lactose) in healthy volunteers ( n=94). Dependent variables were subjective and physiological measures of arousal, as well as serum carisoprodol and caffeine levels. Data were collected from 15 to 280 min after administration of drug or placebo. RESULTS: Caffeine increased alertness, systolic and diastolic blood pressure, startle blink reflexes, and skin conductance responses. Subjects were calmer after carisoprodol, and heart rate was increased. There was a positive correlation between increased arousal and carisoprodol levels when stimulant information had been provided. However, this was only seen when carisoprodol levels were very low. There was no evidence that caffeine modulated the placebo response. CONCLUSIONS: Active placebo responses were seen only transiently when carisoprodol levels were low, and only in the subjective arousal data. Caffeine did not support active placebo responses. The overall findings did not favour the active placebo hypothesis. PMID- 15549278 TI - Effect of short-term SSRI treatment on cognitive bias in generalised anxiety disorder. AB - RATIONALE: There is considerable evidence showing that individuals with generalised anxiety disorder (GAD) selectively process threat-related information, e.g. they have a bias to interpret ambiguous information in a threat related manner. Cognitive theories of anxiety, which provide the basis of cognitive-behaviour therapy, propose that such processing biases play an important role in causing and maintaining anxiety. OBJECTIVES: Given that treatment with selective serotonin re-uptake inhibitors (SSRIs) appears to be effective for GAD, we examined whether it is successful in removing cognitive bias. METHODS: The clinical group included 19 patients with a diagnosis of GAD, and the control group consisted of a non-clinical sample of volunteers, matched for age, gender and years in education. The patients were assessed on measures of interpretative bias (homophone task), anxiety and depression before being prescribed an SSRI (paroxetine or citalopram). After 4 weeks, the cognitive task and mood measures were repeated in the patient group. RESULTS: Prior to treatment, the GAD group showed a significantly greater level of threat-related interpretive bias than controls. Following SSRI treatment, there were significant reductions in both interpretive bias and in anxiety levels in the GAD group. Furthermore, individuals who showed greater clinical improvement (e.g. reflected by reduced anxiety scores) showed a correspondingly greater reduction in their cognitive bias. CONCLUSION: The results suggest that SSRIs are effective in modifying both subjective anxiety levels and threat-related interpretive bias. PMID- 15549279 TI - Motor skill training induces changes in the excitability of the leg cortical area in healthy humans. AB - Training-induced changes in cortical excitability may play an important role in rehabilitation of gait ability in patients with neurological disorders. In this study, we investigated the effect of a 32-min period of motor skill, non-skill and passive training involving the ankle muscles on leg motor cortical excitability in healthy humans. Transcranial magnetic stimulation (TMS) at a range of intensities was applied to obtain a recruitment curve of the motor evoked potentials (MEPs) in the tibialis anterior (TA) muscle before and after training. We also explored the effect of training on inhibitory and facilitatory cortical circuits by using a paired-pulse TMS technique at intervals of 2.5 ms (short-interval intracortical inhibition, SICI) and 8 ms (intracortical facilitation, ICF). During motor skill training, subjects were instructed to make a cursor follow a series of target lines on a computer screen by performing voluntary ankle dorsi- and plantarflexion movements. Non-skill and passive training consisted of repeated voluntary and assisted dorsi- and plantarflexion movements, respectively. Recruitment curves increased significantly after 32 min of motor skill training but not after non-skill and passive training, suggesting that only skill motor training increases motor cortical excitability. Motor skill training was not accompanied by any changes in the recruitment curves of TA MEPs evoked by transcranial electrical stimulation, suggesting that the increased MEPs to TMS was likely caused by changes in excitability at a cortical site. SICI was decreased after 32 min of motor skill training but no changes were observed in ICF. We conclude that similar plastic changes as have previously been reported for the hand motor following motor skill training may also be observed for the leg motor area. The observed plastic changes appeared to be related to the degree of difficulty in the motor task, and may be of relevance for rehabilitation of gait disorders. PMID- 15549280 TI - Initiation of evasive manoeuvres during self-motion: a test of three hypotheses. AB - To understand performance of evasive and interceptive actions it is important to know how people decide when to initiate a movement-initiating at the 'right' moment is often essential for successful performance. It has been proposed that initiation is triggered when a perceptually derived quantity reaches an invariant criterion value. Candidate quantities include time-to-collision (TTC), distance, and rate of image expansion (ROE), all of which have received empirical support. We studied initiation of an evasive manoeuvre in a computer-simulated steering task in which the observer was required to steer through a stationary visual environment and avoid colliding with an obstacle in their path. The results could not be explained by hypotheses which propose that evasive manoeuvre initiation is based on a fixed criterion value of TTC or distance. The overall pattern was, however, consistent with the use of a criterion ROE value. This was further tested by analyses designed to directly evaluate whether the ROE value used to initiate the response was the same across experimental conditions. Only two of the six participants showed evidence for using the ROE strategy. PMID- 15549281 TI - Verbal instructions and top-down saccade control. AB - Few studies have addressed the interaction between instruction content and saccadic eye movement control. To assess the impact of instructions on top-down control, we instructed 20 healthy volunteers to deliberately delay saccade triggering, to make inaccurate saccades or to redirect saccades--i.e. to glimpse towards and then immediately opposite to the target. Regular pro- and antisaccade tasks were used for comparison. Bottom-up visual input remained unchanged and was a gap paradigm for all instructions. In the inaccuracy and delay tasks, both latencies and accuracies were detrimentally impaired by either type of instruction and the variability of latency and accuracy was increased. The intersaccadic interval (ISI) required to correct erroneous antisaccades was shorter than the ISI for instructed direction changes in the redirection task. The word-by-word instruction content interferes with top-down saccade control. Top-down control is a time consuming process, which may override bottom-up processing only during a limited time period. It is questionable whether parallel processing is possible in top-down control, since the long ISI for instructed direction changes suggests sequential planning. PMID- 15549282 TI - Effect of stimulus probability on anti-saccade error rates. AB - Subjects sometimes fail to suppress a reflexive saccade towards the flashed stimulus in an anti-saccade task. Here, we studied how error rates in the anti saccade task varied as a function of saccadic probability. Ten subjects performed 200 anti-saccade trials for each of three saccade-direction probability conditions (20%, 50%, and 80%). We found that as the likelihood of a saccade in a given direction increased, the percentage of pro-saccade errors also increased for stimulus presentations in this direction. These results provide support for the hypothesis that errors in the anti-saccade task are the result of an increased level of motor preparation. PMID- 15549283 TI - Ultrasound features of a paratesticular arteriovenous malformation: a case report of an 11-year-old boy. AB - A paratesticular arteriovenous malformation (AVM) in an 11-year-old boy with a solitary testicle led to a testicle-sparing excision. The radiological features of this rare lesion are reviewed. PMID- 15549284 TI - Coincidence-detection FDG-PET versus gallium in children and young adults with newly diagnosed Hodgkin's disease. AB - Coincidence-detection 18F-FDG-PET (PET) and 67Ga whole-body and SPECT (Ga) were compared in children and young adults with newly diagnosed Hodgkin's disease (HD). MATERIALS AND METHODS: Thirty patients with histologically confirmed HD underwent PET with attenuation correction 1 h after injection of 150-220 MBq 18F FDG and whole-body and SPECT imaging 72 h after injection of 250-370 MBq 67Ga citrate. Two experienced readers retrospectively reviewed PET and Ga scans, grading 13 anatomic regions from one (normal) to five (abnormal). Numerical stages were assigned based on Ann Arbor classification. Comparison was made with disease sites (established by biopsy or two or more of the following: physical examination, conventional imaging studies, radionuclide studies, and follow-up studies) and clinical stages. Sensitivity, specificity, and accuracy were calculated and significance of differences determined using McNemar's test. RESULTS: PET detected 120/138 (87%) disease sites and Ga 109/138 (79%). PET and Ga were concordant for 103/138 (75%) sites. Accuracies were not significantly different for supradiaphragmatic disease. PET was more accurate than Ga for detecting splenic (0.91 vs 0.61, P = 0.012), infradiaphragmatic (0.89 vs 0.75, P = 0.042), and all disease sites combined (0.95 vs 0.91, P = 0.039). PET stage agreed with clinical stage in 79% of patients and Ga in 71%. CONCLUSION: PET was superior to Ga for evaluating children and young adults with newly diagnosed HD. PMID- 15549286 TI - Analysis of the bacterial community inhabiting an aerobic thermophilic sequencing batch reactor (AT-SBR) treating swine waste. AB - The microflora of a self-heating aerobic thermophilic sequencing batch reactor (AT-SBR) treating swine waste was investigated by a combination of culture and culture-independent techniques. The temperature increased quickly in the first hours of the treatment cycles and values up to 72 degrees C were reached. Denaturing gradient gel electrophoresis of the PCR-amplified V3 region of 16S rDNA (PCR-DGGE) revealed important changes in the bacterial community during 3 day cycles. A clone library was constructed with the near-full-length 16S rDNA amplified from a mixed-liquor sample taken at 60 degrees C. Among the 78 non chimeric clones analysed, 20 species (here defined as clones showing more than 97% sequence homology) were found. In contrast to other culture-independent bacterial analyses of aerobic thermophilic wastewater treatments, species belonging to the Bacilli class were dominant (64%) with Bacillus thermocloacae being the most abundant species (38%). The other Bacilli could not be assigned to a known species. Schineria larvae was the second most abundant species (14%) in the clone library. Four species were also found among the 19 strains isolated, cultivated and identified from samples taken at 40 degrees C and 60 degrees C. Ten isolates showed high 16S rDNA sequence homology with the dominant bacterium of a composting process that had not been previously isolated. PMID- 15549287 TI - Bile acids are new products of a marine bacterium, Myroides sp. strain SM1. AB - Strain SM1 was isolated as a biosurfactant-producing microorganism from seawater and presumptively identified as Myroides sp., based on morphology, biochemical characteristics and 16S rDNA sequence. The strain produced surface-active compounds in marine broth, which were purified, using emulsification activity for n-hexadecane as an indicator. The purified compounds were identified by thin layer chromatography, (1)H- and (13)C-NMR spectra and fast atom bombardment mass spectrometry as cholic acid, deoxycholic acid and their glycine conjugates. Type strains of the genus Myroides, M. odoratus JCM7458 and M. odoramitimus JCM7460, also produced these compounds. Myroides sp. strain SM1 possessed a biosynthetic route to cholic acid from cholesterol. Thus, bile acids were found as new products of prokaryotic cells, genus Myroides. PMID- 15549288 TI - Transgenic tobacco expressing fungal laccase promotes the detoxification of environmental pollutants. AB - The phytoremediation of soils contaminated with organic pollutants offers a low cost method for removal of such pollutants. We have attempted to enhance the environmental decontamination functions of plants by introducing appropriate enzymatic activities from microorganisms. In the present study, we introduced an extracellular fungal enzyme, the laccase of Coriolus versicolor, into tobacco plants. One transgenic plant, designated FL4, produced laccase that was secreted into the rhizosphere. FL4 was able to remove 20 micromol bisphenol A or pentachlorophenol per gram dry weight. The efficiency of this removal was apparently greater than that of control lines. Our results should stimulate efforts to develop plant-based technologies for the removal of environmental pollutants from contaminated environments. PMID- 15549289 TI - Phosphonium ionic liquids for degradation of phenol in a two-phase partitioning bioreactor. AB - Six ionic liquids (ILs), which are organic salts that are liquid at room temperature, were tested for their biocompatibility with three xenobiotic degrading bacteria, Pseudomonas putida, Achromobacter xylosoxidans, and Sphingomonas aromaticivorans. Of the 18 pairings, seven were found to demonstrate biocompatibility, with one IL (trihexyl(tetradecyl)phosphonium bis(trifluoromethylsulfonyl) amide) being biocompatible with all three organisms. This IL was then used in a two-phase partitioning bioreactor (TPPB), consisting of 1 l aqueous phase loaded with 1,580 mg phenol and 0.25 l IL, inoculated with the phenol degrader P. putida. This initially toxic aqueous level of phenol was substantially reduced by phenol partitioning into the IL phase, allowing the cells to utilize the reduced phenol concentration. The partitioning of phenol from the IL to the aqueous phase was driven by cellular demand and thermodynamic equilibrium. All of the phenol was consumed at a rate comparable to that of previously used organic-aqueous TPPB systems, demonstrating the first successful use of an IL with a cell-based system. A quantitative (31)P NMR spectroscopic assay for estimating the log P values of ILs is under development. PMID- 15549290 TI - Influence of the carbon/nitrogen/phosphorus ratio on polycyclic aromatic hydrocarbon degradation by Mycobacterium and Sphingomonas in soil. AB - Biodegradation of polycyclic aromatic hydrocarbons (PAHs) in the environment is often limited due to unfavorable nutrient conditions for the bacteria that use these PAHs as sole source of carbon and energy. Mycobacterium and Sphingomonas are 2 PAH-degrading specialists commonly present in PAH-polluted soil, but not much is known about their specific nutrient requirements. By adding different inorganic supplements of nitrogen (N) and phosphorus (P), affecting the overall carbon/nitrogen/phosphorus ratio of soil in soil slurry degradation tests, we investigated the impact of soil inorganic N and P nutrient conditions on PAH degradation by PAH-degrading Sphingomonas and Mycobacterium strains. The general theoretically calculated C/N/P ratio of 100/10/1 (expressed in moles) allowed rapid PAH metabolization by Sphingomonas and Mycobacterium strains without limitation. In addition, PAH-degradation rate and extent was not affected when ca. ten times lower concentrations of N and P were provided, indicating that Sphingomonas and Mycobacterium strains are capable of metabolizing PAHs under low nutrient conditions. Nor does PAH-degradation seem to be affected by excesses of N and P creating an imbalanced C/N/P ratio. However, supplements of N and P salts increased the salinity of soil slurry solutions and seriously limited or even completely blocked biodegradation. PMID- 15549291 TI - Enantioselective reduction of carbonyl compounds by whole-cell biotransformation, combining a formate dehydrogenase and a (R)-specific alcohol dehydrogenase. AB - A whole-cell biotransformation system for the reduction of prochiral carbonyl compounds, such as methyl acetoacetate, to chiral hydroxy acid derivatives [methyl (R)-3-hydroxy butanoate] was developed in Escherichia coli by construction of a recombinant oxidation/reduction cycle. Alcohol dehydrogenase from Lactobacillus brevis catalyzes a highly regioselective and enantioselective reduction of several ketones or keto acid derivatives to chiral alcohols or hydroxy acid esters. The adh gene encoding for the alcohol dehydrogenase of L. brevis was expressed in E. coli. As expected, whole cells of the recombinant strain produced only low quantities of methyl (R)-3-hydroxy butanoate from the substrate methyl acetoacetate. Therefore, the fdh gene from Mycobacterium vaccae N10, encoding NAD+-dependent formate dehydrogenase, was functionally coexpressed. The resulting two-fold recombinant strain exhibited an in vitro catalytic alcohol dehydrogenase activity of 6.5 units mg-1 protein in reducing methyl acetoacetate to methyl (R)-3-hydroxy butanoate with NADPH as the cofactor and 0.7 units mg-1 protein with NADH. The in vitro formate dehydrogenase activity was 1.3 units mg-1 protein. Whole resting cells of this strain catalyzed the formation of 40 mM methyl (R)-3-hydroxy butanoate from methyl acetoacetate. The product yield was 100 mol% at a productivity of 200 micromol g-1 (cell dry weight) min-1. In the presence of formate, the intracellular [NADH]/[NAD+] ratio of the cells increased seven-fold. Thus, the functional overexpression of alcohol dehydrogenase in the presence of formate dehydrogenase was sufficient to enable and sustain the desired reduction reaction via the relatively low specific activity of alcohol dehydrogenase with NADH, instead of NADPH, as a cofactor. PMID- 15549292 TI - Hydroxylation activity of P450 BM-3 mutant F87V towards aromatic compounds and its application to the synthesis of hydroquinone derivatives from phenolic compounds. AB - Cytochrome P450 BM-3 from Bacillus megaterium is a fatty acid hydroxylase exhibiting selectivity for long-chain substrates (12-20 carbons). Replacement of Phe87 in P450 BM-3 by Val (F87V) greatly increased its activity towards a variety of aromatic and phenolic compounds. The apparent initial reaction rates of F87V as to benzothiophene, indan, 2,6-dichlorophenol, and 2-(benzyloxy)phenol were 227, 204, 129, and 385 nmol min(-1) nmol(-1) P450, which are 220-, 66-, 99-, and 963-fold those of the wild type, respectively. These results indicate that Phe87 plays a critical role in the control of the substrate specificity of P450 BM-3. Furthermore, F87V catalyzed regioselective hydroxylation at the para position of various phenolic compounds. In particular, F87V showed high activity as to the hydroxylation of 2-(benzyloxy)phenol to 2-(benzyloxy)hydroquinone. With F87V as the catalyst, 0.71 mg ml(-1) 2-(benzyloxy)hydroquinone was produced from 1.0 mg ml(-1) 2-(benzyloxy)phenol in 4 h, with a molar yield of 66%. PMID- 15549293 TI - Purification and characterization of recombinant Escherichia coli-expressed Pichia etchellsii beta-glucosidase II with high hydrolytic activity on sophorose. AB - Beta-glucosidase II (Bgl II), encoded by the betaglu2 gene of the thermo-tolerant yeast Pichia etchellsii, was purified from recombinant Escherichia coli pBG22:JM109. The enzyme had a molecular mass of 176 kDa and was a dimer with an apparent subunit mass of 83 kDa. It exhibited broad substrate specificity and hydrolyzed beta-linked gluco-disaccharides and oligosaccharides, salicin, and cyanogenic glucoside amygladin. The unusually high hydrolytic activity of 7,680 units min(-1) g(-1) protein was obtained on sophorose. Competition experiments performed using differently linked beta-disaccharides indicated these to be hydrolyzed at the same active site. Transglycosylation activity leading to the biosynthesis of several disaccharides and oligosaccharides was observed. The enzyme was placed in glycosyl hydrolase family 3, based on a statistical approach using amino acid composition data. The involvement of His as a catalytically important residue was confirmed by diethylpyrocarbonate modification. Pre incubation of the purified enzyme with 5 mM p-nitrophenyl-beta-D-glucoside offered 2.5-fold higher residual activity compared with unbound enzyme, indicating protection at the active site. The feasibility of this enzyme as a biocatalyst of choice for the synthesis of glyco-conjugates is discussed. PMID- 15549294 TI - 18FDG-PET for monitoring disease activity in an HIV-1 positive patient with disseminated chronic osteomyelitic brucellosis due to Brucella melitensis. PMID- 15549295 TI - PET scan with 11C-acetate for the imaging of liver masses: report of a false positive case. PMID- 15549296 TI - The detection of aortic valve infection by FDG-PET/CT in a patient with infection following total knee replacement. PMID- 15549297 TI - FDG-PET is able to detect pancreatic carcinoma in chronic pancreatitis. AB - PURPOSE: 2-(18F)-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) is an imaging technique which enables detection of malignancies. FDG-PET has been established as a tool for the diagnosis of pancreatic carcinoma (CA). Early detection is mandatory as cure can only be achieved in non-advanced disease. This is, however, very difficult with conventional radiological techniques. Patients with chronic pancreatitis (CP) are at risk of developing CA. A simple, reliable screening method for malignant degeneration is highly desirable. We set out to investigate whether FDG-PET is able to detect CA in the setting of CP and can fulfil a potential role in the early detection of CA in CP. METHODS: FDG-PET was performed in 77 CP patients and in six patients with CP complicated by CA (CP + CA). We included 26 CA patients as a positive control. A positive scan was defined as focal FDG accumulation in the pancreas region. RESULTS: In 67 of the 77 CP (87%) patients, pancreatic FDG accumulation was absent. Six patients had significant accumulation. In CA, 24/26 patients had a positive PET. Five out of the six patients with CP + CA had focal uptake, while minor uptake was seen in one patient. FDG-PET was positive in almost all CA patients and CP + CA patients. FDG-PET was negative in the large majority (87%) of CP patients, which suggests that a positive PET scan in CP patients must lead to efforts to exclude a malignancy. CONCLUSION: These data suggest that FDG-PET has a potential role as a diagnostic tool for detecting CA in longstanding CP. PMID- 15549298 TI - In vivo imaging of adenosine A2A receptors in rat and primate brain using [11C]SCH442416. AB - PURPOSE: The aim of this study was to evaluate the suitability of [11C]SCH442416 for the in vivo imaging of adenosine A2A receptors. METHODS: In rats and Macaca nemestrina, we evaluated the time course of the cerebral distribution of [11C]SCH442416. Furthermore, in rats we investigated the rate of metabolic degradation, the inhibitory effects of different drugs acting on adenosine or dopamine receptors and the modification induced by the intrastriatal administration of quinolinic acid (QA). RESULTS: The rate of metabolic degradation of [11C]SCH442416 in rats was slow; 60 min after tracer injection, more than 40% of total plasma activity was due to unmetabolised [11C]SCH442416. At the time of maximum uptake, radioactive metabolites represented only 6% of total extractable activity in the cerebellum and less than 1% in the striatum. In the striatum, the region with the highest expression of A2A receptors, the in vivo uptake of [11C]SCH442416 was significantly reduced only by drugs acting on A2A receptors or by QA, a neurotoxin that selectively reduces the number of intrastriatal GABAergic neurons. Position emission tomography (PET) studies in monkeys indicated that the tracer rapidly accumulates in brain, reaching maximum uptake between 5 and 10 min. Twenty minutes after the injection, radioactivity concentration in the striatum was two times that in the cerebellum. CONCLUSION: The specificity of binding, the rank order of regional distribution in the brain of rats and M. nemestrina, the good signal to noise ratios and the low amount of radioactive metabolites in brain and periphery indicate that [11C]SCH442416 is a promising tracer for the in vivo imaging of A2A adenosine receptors using PET. PMID- 15549299 TI - Re: Percutaneous plating of distal tibial fractures (2004) 28(3):159. PMID- 15549300 TI - The abductor pollicis longus: relation between innervation, muscle bellies and number of tendinous slips. AB - The aim of this study was to assess the relation between the insertions of the distal tendinous slips, the muscle bellies and the innervation pattern of the abductor pollicis longus (APL) muscle and of the extensor pollicis brevis (EPB). The upper extremities of 31 frozen cadavers were dissected under magnifying lenses to describe the distribution of the posterior interosseous nerve (PION). The number and the distribution of distal tendinous slip insertions of the APL muscle were variable. Two superficial and deep distal tendon groups were noted. The separation into superficial and deep muscular parts of the APL was frequent (87%). The EPB muscle was generally constituted by one muscle belly and one tendinous slip (93.5%). The innervation by the PION to the APL and EPB muscles was classified into five types. The specific innervation between superficial and deep muscular parts of the APL muscle, the specific innervation of the deep muscle bellies and the independence of the superficial and deep distal tendon groups of the APL muscle are arguments in favor of a complex functional role of the APL motor unit in thumb mechanics. However, no independence of the tendinous slips in the two distal tendon groups and no correlation between the number of tendinous slips and muscle bellies or innervation were observed. These limit the functional role of the two independent superficial and deep musculotendinous APL motor units. The use of the APL tendon for interposition arthroplasty, for tendon transfer or tendon translocation seems logical, particularly if using one of the two distal tendon groups. PMID- 15549301 TI - The anatomical basis of the osteo-musculo-cutaneous trapezius flap in mandibular reconstruction. AB - The osteo-musculo-cutaneous trapezius flap has seen its indications in mandibular reconstruction reduced since the appearance of micro-anastomotic flaps. However, it may still be useful when the patient has marked atheroma contraindicating a free flap. While the arterial supply of this flap is well known, the same is not true for its venous drainage. The preservation of the accessory nerve and its limits in mandibular reconstruction has been little studied. We carried out a study on 30 subjects (60 cadaveric dissections of trapezius flaps) in order to address these questions. The study has been completed by a surgical series of five patients. The cadaveric study allowed demonstration of the variability of venous drainage of this flap, which depended on the external jugular vein in 80% of cases, the subclavian vein in 12% of cases and on both veins in about 8% of cases. The accessory nerve in one third of cases passed through the middle of the arteriovenous pedicle making its preservation impossible. The segment of the scapular spine allowed reconstruction of about 9 cm of mandible including the mental protuberance in 95% of cases. The surgical study confirmed the data of the cadaveric study and showed the value of this flap when free flaps are contraindicated. PMID- 15549302 TI - Reduction of catheter-related infections in neutropenic patients: a prospective controlled randomized trial using a chlorhexidine and silver sulfadiazine impregnated central venous catheter. AB - Antiseptic coating of intravascular catheters may be an effective means of decreasing catheter-related colonization and subsequent infection. The purpose of this study was to assess the efficacy of chlorhexidine and silver sulfadiazine (CH-SS)-impregnated central venous catheters (CVCs) to prevent catheter-related colonization and infection in patients with hematological malignancies who were subjected to intensive chemotherapy and suffered from severe and sustained neutropenia. Proven CVC-related bloodstream infection (BSI) was defined as the isolation of the same species from peripheral blood culture and CVC tip (Maki technique). This randomized, prospective clinical trial was carried out in 106 patients and compared catheter-related colonization and BSI using a CH-SS impregnated CVC (n=51) to a control arm using a standard uncoated triple-lumen CVC (n=55). Patients were treated for acute leukemia (n=89), non-Hodgkin's lymphoma (n=10), and multiple myeloma (n=7). Study groups were balanced regarding to age, sex, underlying diseases, insertion site, and duration of neutropenia. The CVCs were in situ a mean of 14.3+/-8.2 days (mean+/-SD) in the study group versus 16.6+/-9.7 days in the control arm. Catheter-related colonization was observed less frequently in the study group (five vs nine patients; p=0.035). CVC related BSI were significantly less frequent in the study group (one vs eight patients; p=0.02). In summary, in patients with severe neutropenia, CH-SS impregnated CVCs yield a significant antibacterial effect resulting in a significantly lower rate of catheter-related colonization as well as CVC-related BSI. PMID- 15549303 TI - Non-myeloablative stem cell transplantation for autoimmune diseases. AB - Treatment of life-threatening autoimmune diseases in animal models with induced or spontaneous autoimmune diseases can be accomplished by a 2-step procedure involving elimination of self-reactive lymphocytes with an immune ablative conditioning regimen followed by infusion of autologous or allogeneic stem cells, respectively. In animal models it was shown that using such a strategy, autoimmunity could be adequately controlled. It is speculated that de-novo development of the T and B cell repertoire from uncommitted progenitor cells in the presence of the autoantigens may be the best recipe for re-induction of self tolerance, similarly to the normal ontogeny of the immune system during the induction of self tolerance in fetal stage. For both autologous and allogeneic hematopoietic stem cell transplantation, a non-myeloablative stem cell transplantation (NST) regimen may be used for safer lymphoablation rather than myeloablation. In addition, for allogeneic hematopoietic stem cell transplantation engraftment of disease resistant donor stem cells will alter the genetic predisposition towards autoimmune disease susceptibility. PMID- 15549304 TI - Allogeneic hematopoietic cell transplantation as treatment for hematological malignancies: a review. AB - Allogeneic hematopoietic cell transplantation (HCT) was originally developed as a form of rescue from high-dose chemoradiotherapy, which is given both to eradicate malignancy and provide sufficient immunosuppression for allogeneic engraftment. The first attempts of allogeneic HCT in humans met with little success. However, a better understanding of the complexities of the human leukocyte antigen (HLA) system has allowed selecting compatible sibling donors, and the development of postgrafting immunosuppressive regimens has helped prevent serious graft-versus host disease, thereby changing the role of allogeneic HCT from a desperate therapeutic maneuver to a curative treatment modality for many patients with malignant hematological diseases. In addition, the establishment of large registries of HLA-typed volunteers has permitted finding suitable unrelated donors for many patients without family donors. Further advances in the immunogenetics of HLA, especially typing by molecular techniques, have improved results after unrelated HCT, which have begun resembling those obtained with HLA identical sibling grafts, at least in young patients. Important advances have also been made in the prevention and treatment of infectious complications and in other areas of supportive care. Since the late seventies, it has been recognized that allogeneic immunocompetent cells transplanted with the stem cells, or arising from them, mediated therapeutic anti-tumor effects independent of the action of the high-dose therapy, termed graft-versus-tumor (GVT) effects. This has prompted the recent development of non-myeloablative conditioning regimens for allogeneic HCT that have opened the way to include elderly patients and those with comorbid conditions. Remaining challenges include further advances in the prevention and treatment of both severe graft-versus-host disease and infections. Also, progress in adoptive transfer of T cells with relative tumor specificity and disease-targeted therapy with agents such as Imatinib, Rituximab or radiolabeled monoclonal antibodies would make allogeneic HCT even more effective. PMID- 15549305 TI - Allogeneic stem cell transplantation for the treatment of diseases associated with a deficiency in bone marrow products. AB - Our understanding of the pathophysiology of hematopoietic failure associated syndromes led to the developmental of potentially curative procedures for the treatment of many diseases including Severe aplastic anemia, Fanconi's anemia, Primary immunodeficiency, Osteopetrosis, and Metabolic diseases. Although the number of patients that were transplanted for bone marrow deficiency diseases is relatively low as compared to patients with hematological malignancies, the impact on the knowledge of hematopoiesis and transplantation biology is tremendous. Moreover, the patient's average young age suffering from these diseases further encourage searching for curative approaches. Lucking a fully MHC matched donor, remained a significant obstacle in stem cell transplantation for non-malignant hematological disorders. Lessons from attempts to cure aplasic anemia with bone marrow transplantation guided us to the improvement of pretransplant conditioning regimens and prevention of graft versus host reactions after transplantation. Furthermore, in recent years optimization of disease specific protocol have been successfully designed and clinically applied. PMID- 15549306 TI - Cord blood transplant: strategy of alternative donor search. AB - Unrelated cord blood transplantation has been used to treat patients with malignant and non-malignant hematopoietic disorders for whom an HLA-compatible hematopoietic stem cell donor is not available. The establishment of cord blood banks worldwide, the increased number of cord blood units frozen, and the shorter time to find a donor, have made it possible to use this source of hematopoietic stem cells to treat more than 2,500 patients. The Eurocord registry was established to study the clinical results of cord blood transplantation and to compare the outcomes of unrelated transplants using either cord blood or bone marrow. Briefly, we have found, in two distinct retrospective analyses of children or adults with acute leukemia given either an unrelated cord blood or bone marrow transplant, that leukemia-free survival and relapse were similar in both types of graft (with adjustment for confounding clinical factors). Cord blood recipients experienced a decreased incidence of acute graft-versus-host disease and delayed hematopoietic recovery compared to bone marrow recipients. To improve the delayed hematopoietic recovery after cord blood transplantation, certain approaches have been investigated such as ex vivo expansion of cord blood cells, double cord blood transplantation and reduced intensity conditioning regimen. We have also attempted to establish some guidelines for cord blood-donor choice based on cord blood cell dose and number of HLA disparities that have been found to be associated with hematopoietic recovery. In conclusion, an unrelated hematopoietic stem cell donor should be simultaneously searched for in cord blood banks and in bone marrow donor registries for patients lacking an HLA-identical sibling hematopoietic stem cell donor. The option of performing cord blood transplants should be based on urgency of the transplant, cord blood cell dose and number of HLA disparities. PMID- 15549308 TI - Structural identifiability analysis of some highly structured families of statespace models using differential algebra. AB - In this paper we identify biologically relevant families of models whose structural identifiability analysis could not be performed with available techniques directly. The models considered come from both the immunological and epidemiological literature. PMID- 15549309 TI - Multistage carcinogenesis and the fraction at risk. AB - Multistage carcinogenesis models describe the evolution of the cells in an individual's organ from a normal stage to a pre-neoplastic stage to a neoplastic stage. The triggers for the passage from one stage to the next one are presumed to be genetic alterations, which are not only governed by purely random events but also by individual environmental and genetic factors. We generalize existing models of carcinogenesis to populations composed of heterogeneous individuals, thus taking the environmental and genetic variability into account. PMID- 15549307 TI - Immunogenicity of human embryonic stem cells: can we achieve tolerance? AB - Human embryonic stem cells are unique in their capacity to propagate without losing pluripotency, and at the same time may readily differentiate to various cell types of the three embryonic germ layers. It is widely accepted today that differentiated human embryonic stem cells may in the future enable repair of vital tissues of the body. Detailed differentiation protocols need to be developed and safety issues associated with cellular therapeutics must be examined. One of the greatest hurdles facing transplantation is the development of immune rejection processes towards non-autologous cells. Profiling of histocompatibility antigens expressed on the cells reveals that they might be subjected to immune response. Here we describe the routes of immune recognition that can identify these antigens and the proposed ways for overcoming the rejection of human embryonic stem cell derivatives. PMID- 15549310 TI - Convergence of a structured metapopulation model to Levins's model. AB - We consider a structured metapopulation model describing the dynamics of a single species, whose members are located in separate patches that are linked through migration according to a mean field rule. Our main aim is to find conditions under which its equilibrium distribution is reasonably approximated by that of the unstructured model of Levins (1969). We do this by showing that the (positive) equilibrium distribution converges, as the carrying capacity of each population goes to infinity together with appropriate scalings on the other parameters, to a bimodal distribution, consisting of a point mass at 0, together with a positive part which is closely approximated by a shifted Poisson centred near the carrying capacity. Under this limiting regime, we also give simpler approximate formulae for the equilibrium distribution. We conclude by showing how to compute persistence regions in parameter space for the exact model, and then illustrate all our results with numerical examples. Our proofs are based on Stein's method. PMID- 15549311 TI - Dynamic heterogeneous spatio-temporal pattern formation in host-parasitoid systems with synchronised generations. AB - In this paper we develop a general mathematical model describing the spatio temporal dynamics of host-parasitoid systems with forced generational synchronisation, for example seasonally induced diapause. The model itself may be described as an individual-based stochastic model with the individual movement rules derived from an underlying continuum PDE model. This approach permits direct comparison between the discrete model and the continuum model. The model includes both within-generation and between-generation mechanisms for population regulation and focuses on the interactions between immobile juvenile hosts, adult hosts and adult parasitoids in a two-dimensional domain. These interactions are mediated, as they are in many such host-parasitoid systems, by the presence of a volatile semio-chemical (kairomone) emitted by the hosts or the hosts' food plant. The model investigates the effects on population dynamics for different host versus parasitoid movement strategies as well as the transient dynamics leading to steady states. Despite some agreement between the individual and continuum models for certain motility parameter ranges, the model dynamics diverge when host and parasitoid motilities are unequal. The individual-based model maintains spatially heterogeneous oscillatory dynamics when the continuum model predicts a homogeneous steady state. We discuss the implications of these results for mechanistic models of phenotype evolution. PMID- 15549312 TI - An algorithm for molecular dissection of tumor progression. AB - The volumetric growth of tumor cells as a function of time is most often likely to be a complex trait, controlled by the combined influences of multiple genes and environmental influences. Genetic mapping has proven to be a powerful tool for detecting and identifying specific genes affecting complex traits, i.e., quantitative trait loci (QTL), based on polymorphic markers. In this article, we present a novel statistical model for genetic mapping of QTL governing tumor growth trajectories in humans. In principle, this model is a combination of functional mapping proposed to map function-valued traits and linkage disequilibrium mapping designed to provide high resolution mapping of QTL by making use of recombination events created at a historic time. We implement an EM simplex hybrid algorithm for parameter estimation, in which a closed-form solution for the EM algorithm is derived to estimate the population genetic parameters of QTL including the allele frequencies and the coefficient of linkage disequilibrium, and the simplex algorithm incorporated to estimate the curve parameters describing the dynamic changes of cancer cells for different QTL genotypes. Extensive simulations are performed to investigate the statistical properties of our model. Through a number of hypothesis tests, our model allows for cutting-edge studies aimed to decipher the genetic mechanisms underlying cancer growth, development and differentiation. The implications of our model in gene therapy for cancer research are discussed. PMID- 15549313 TI - Deterministic and stochastic modelling of endosome escape by Staphylococcus aureus: "quorum" sensing by a single bacterium. AB - Deterministic and stochastic models describing quorum sensing by Staphylococcus aureus within an endosome, and the subsequent escape via the production of virulence factors, are developed and analysed. Particular attention is given to a biologically-relevant asymptotic limit of the problem, for which the solutions, including the endosome escape time, can be explicitly characterised in terms of the model parameters. PMID- 15549314 TI - Information and effective number of meioses in linkage analysis. AB - In this paper we introduce two information criteria in linkage analysis. The setup is a sample of families with unusually high occurrence of a certain inheritable disease. Given phenotypes from all families, the two criteria measure the amount of information inherent in the sample for 1) testing existence of a disease locus harbouring a disease gene somewhere along a chromosome or 2) estimating the position of the disease locus. Both criteria have natural interpretations in terms of effective number of meioses present in the sample. Thereby they generalize classical performance measures directly counting number of informative meioses. Our approach is conditional on observed phenotypes and we assume perfect marker data. We analyze two extreme cases of complete and weak penetrance models in particular detail. Some consequences of our work for sampling of pedigrees are discussed. For instance, a large sibship family with extreme phenotypes is very informative for linkage for weak penetrance models, more informative than a number of small families of the same total size. PMID- 15549315 TI - [Regression of germ cell tumors after chemotherapy]. AB - Today the treatment of gonadal germ cell tumors is standardized. The cisplatin containing chemotherapy and the multi-modal therapy strategies have increased the rate of successful treatment enormously. Germ cell tumors are almost always treated surgically. Following the rare, primary chemotherapy, the residual tumor must be classified according to the WHO as accurately as possible. A binding system for the documentation of tumor regression does not exist. The diagnostic retroperitoneal lymphadenectomy is also rare. Here as well, the classification is performed according to the WHO and the TNM classification. The examination of the tissue samples from a retroperitoneal lymphadenectomy after chemotherapy is problematic. The morphology is often bizarre, preparatory and terminological standards do not exist. Is there still vital tumor present then it can most often be diagnosed as a teratoma. In that case a classification takes place as to whether it is "mature" or "immature". If a tissue sample contains other differentiations, the classification is performed in detail according to the WHO classification of germ cell tumors. Sarcomas or carcinomas must be reliably distinguished and classified, as they lead to different therapeutic consequences. The terminology must be defined in a binding manner between both the pathology and the clinic, due to the lack of global definitions. PMID- 15549316 TI - A mitochondrial plasmid and plasmid-like RNA and DNA polymerases encoded within the mitochondrial genome of carrot (Daucus carota L.). AB - The mitochondrial genome of mitochondrial type (mitotype) SW3 of carrot (Daucus carota L.) encodes intact reading frames for a RNA polymerase (Rpo) and a DNA polymerase (Dpo) similar to those encoded by linear mitochondrial plasmids from plants. A BLAST search of translated nucleotide sequences in GenBank revealed previously unreported plasmid-like Rpo or Dpo sequence fragments in many plant mitochondrial DNAs. Phylogenetic analyses of the relationships between mitochondrial (mt)DNA-encoded and plasmid-encoded Rpos and Dpos from plants suggest that the mitochondrial sequences were derived from integrated plant plasmid sequences. A linear mitochondrial plasmid was detected in a different mitotype (FG21) of carrot by Southern hybridization of the Rpo and Dpo to undigested mtDNAs. Transcripts of the mtDNA-encoded Rpo and Dpo in mitotype SW3 were detected by RT-PCR. PMID- 15549317 TI - The Gpmk1 MAP kinase of Fusarium graminearum regulates the induction of specific secreted enzymes. AB - Recently, we described Gpmk1 MAP kinase-disruption mutants of Fusarium graminearum that were fully viable in vitro, but had completely lost their ability to infect wheat. As cell wall-degrading enzymes are postulated to participate in the infection process of F. graminearum, these MAP kinase disruption mutants were analysed for their ability to produce cell wall-degrading enzymes in vitro and compared with the wild-type strain. The gpmk1 disruption had no effect on the production of pectinolytic or amylolytic enzymes. However, Gpmk1 regulates the early induction of extracellular endoglucanase, xylanolytic, and proteolytic activities. Furthermore, the MAP kinase was responsible for the overall induction of secreted lipolytic activities. Since the disruption of the Gpmk1 MAP kinase leads to an apathogenic phenotype, these results suggest that the infection process of F. graminearum depends on the secretion of cell wall degrading enzymes, particularly during the early infection stage. Finally, this work provides the first detailed analysis of the apathogenic phenotype of the F. graminearum Gpmk1 mutants. PMID- 15549318 TI - Microsatellite markers reveal genetic differentiation among populations of Sclerotinia sclerotiorum from Australian canola fields. AB - Eight microsatellite markers were applied to 154 Sclerotinia sclerotiorum isolates from four Australian canola fields, to determine the extent of genetic variation and differentiation in populations of this pathogen. A total of 82 different haplotypes were identified and in each population many haplotypes were unique. Mycelial compatibility grouping, a phenotypic marker system controlled by multiple loci, was often associated with groups of identical or closely related microsatellite haplotypes. Genotypic diversity ranged from 36% to 80% of maximum in the four populations, and gene diversity ranged from 0.23 to 0.79. Genotypic disequilibrium analyses on each of the four populations suggested that both clonal and sexual reproduction contributed to population structure. Analyses based on genetic diversity and fixation indices demonstrated a moderate to high level of differentiation (R(ST)=0.16-0.33, F(ST)=0.18-0.23) between populations from New South Wales and those from Victoria. Despite this genetic diversity, most isolates did not vary in virulence on canola leaves. PMID- 15549319 TI - Random mutagenesis of an essential Candida albicans gene. AB - A method for the analysis of Candida albicans gene function, which involves random mutagenesis of the open reading frame, is described. This method is especially suited for the study of essential and multi-functional genes, with several advantages over regulatable promoters more commonly used to study essential gene function. These advantages include expression from the endogenous promoter, which should yield a more appropriate transcript expression and abrogate the need for shifts in carbon or amino acid sources necessary with the use of regulatable promoters. Furthermore, there is potential for isolating individual functions of multi-functional genes. To verify this experimental approach, we randomly mutated the essential C. albicans gene, BMH1. The resulting "pool" of putative mutant alleles was then introduced into a BMH1/bmh1Delta strain of C. albicans, such that only the mutagenized BMH1 sequences could be expressed. Transformants were screened for rapamycin sensitivity, defects in filamentation on M199 agar, and growth at 42 degrees C. In this way, we identified six non-lethal mutant alleles of BMH1 with altered amino acid sequences. Further phenotypic analysis of these mutant strains enabled us to segregate individual functions of C. albicans BMH1. The relative merits of Escherichia coli versus PCR-mediated mutagenesis are discussed. PMID- 15549320 TI - CT measurement of coronary calcium mass: impact on global cardiac risk assessment. AB - Coronary calcium mass percentiles can be derived from electron beam CT as well as from multidetector-row CT of all manufacturers. Coronary calcium mass may serve as a more individualized substitute for age for cardiac risk stratification. The aim was to investigate the potential impact of CT coronary calcium mass quantification on cardiac risk stratification using an adjusted Framingham score. Standardized coronary calcium mass was determined by multidetector-row CT in a total of 1,473 patients (1,038 male, 435 female). The impact on risk stratification of replacing the traditional Framingham age point score by a point score based on calcium mass relative to age was tested. Any coronary calcium found in males in the age group of 20-34 years and females in the age group of 20 59 years results in an increase of the Framingham score by 9 and 4-7 points, respectively. Only in males 65 years of age and older, none or minimal amounts of coronary calcium decrease the Framingham score by three points. The coronary calcium mass and age-related scoring system may have impact on the reassignment of patients with an intermediate Framingham risk to a lower or higher risk group. PMID- 15549321 TI - Gastrointestinal complications of bariatric Roux-en-Y gastric bypass surgery. AB - Obesity is rapidly becoming the most important public health issue in USA and Europe. Roux-en-Y gastric bypass is now established as the gold standard for treating intractable morbid or super obesity. We reviewed the imaging findings following this surgery in 234 patients. In this pictorial essay we present the CT and upper gastrointestinal contrast study appearances of the expected postoperative anatomy as well as a range of abdominal complications. The complications are classified into leaks, fistula and obstruction. Postoperative gastric outlet and small bowel obstruction can be caused by anastomotic stenosis, mesocolic tunnel stenosis, adhesions, stomal ulcer, obturation, intussusception and internal or external hernia. Small bowel obstruction may be of a simple, closed loop and/or strangulating type. The radiologist should be able to diagnose the type and possible cause of obstruction. PMID- 15549322 TI - Three dimensional ultrasound and prostate cancer. AB - Three-dimensional ultrasound (3-D US) is a non-invasive method of producing whole volume images of solid structures. Early work on prostate imaging identified several advantages over 2-D imaging with a good ability to detect intraprostatic lesions. Several 3-D transrectal ultrasound (3-D TRUS) systems are now available for prostate imaging. Initial work using gray scale ultrasound appears promising with reported overall staging accuracies of up to 94%. These results were favourable when compared to other modalities for local staging of prostate cancer. Several adjuncts to 3-D gray scale TRUS have been investigated. A greater sensitivity for cancer detection has been achieved with the addition of power colour Doppler and contrast agents. Further clinical applications for 3-D TRUS include assessing placement of brachytherapy seeds and for cyroablation techniques. Computer enhancement with image registration has shown that 3-D US images can be manipulated to derive more information. Although the results of gray scale imaging alone or with adjuncts and post processing appear promising, these techniques remain largely experimental. PMID- 15549323 TI - [Reduction of lipid peroxidation and apoptosis in corneal endothelial cells by vitamin A]. AB - PURPOSE: The goal of this study was to determine the effects of lipid peroxidation-mediated toxicity of iron ions on corneal endothelial cells leading to apoptosis. METHODS: Murine corneal endothelial cells were maintained in tissue culture medium supplemented with free iron ions, known to lead to increased lipid peroxidation. Retinoic acid in the cell supernatant and cytoplasm of these cells was determined using HPLC. The rate of apoptosis was assessed by quantification of caspase-3-like activity. The lipid peroxidation was measured using the malondialdehyde method. Supplementation of retinoic acid was tested in the setting of apoptosis. RESULTS: Free iron ions led to a rapid loss of retinoic acid in the supernatant and the corneal endothelial cells. This was correlated with rising levels of malondialdehyde following oxidative stress and increased apoptosis. Supplementation of retinoic acid alone significantly reduced oxidative stress and apoptosis in the respective cells. CONCLUSION: In this study the authors present a novel in vitro model to test the direct influence of pro oxidative species on corneal endothelial cells. The authors also prove that supplementing corneal endothelial cells with retinoic acid sufficiently prevents free radical injury and apoptosis. PMID- 15549325 TI - Temporal shifts in visual pigment absorbance in the retina of Pacific salmon. AB - The visual pigments and photoreceptor types in the retinas of three species of Pacific salmon (coho, chum, and chinook) were examined using microspectrophotometry and histological sections for light microscopy. All three species had four cone visual pigments with maximum absorbance in the UV (lambda(max): 357-382 nm), blue (lambda(max): 431-446 nm), green (lambda(max): 490-553 nm) and red (lambda(max): 548-607 nm) parts of the spectrum, and a rod visual pigment with lambda(max): 504-531 nm. The youngest fish (yolk-sac alevins) did not have blue visual pigment, but only UV pigment in the single cones. Older juveniles (smolts) had predominantly single cones with blue visual pigment. Coho and chinook smolts (>1 year old) switched from a vitamin A1- to a vitamin A2 dominated retina during the spring, while the retina of chum smolts and that of the younger alevin-to-parr coho did not. Adult spawners caught during the Fall had vitamin A2-dominated retinas. The central retina of all species had three types of double cones (large, medium and small). The small double cones were situated toward the ventral retina and had lower red visual pigment lambda(max) than that of medium and large double cones, which were found more dorsally. Temperature affected visual pigment lambda(max) during smoltification. PMID- 15549326 TI - Oral budesonide therapy improves quality of life in patients with collagenous colitis. AB - INTRODUCTION: Collagenous colitis is an idiopathic microscopic colitis characterised by watery diarrhoea. The impact of collagenous colitis on quality of life has not been assessed. Our aim was to assess quality of life in patients with this condition and compare the effect of treatment with budesonide capsules or placebo on this parameter. METHODS: Patients with chronic diarrhoea and histologically-proven collagenous colitis were randomised to receive either budesonide controlled-release capsules (Entocort capsules, AstraZeneca, Lund, Sweden), 9 mg/day, or placebo for 6 weeks. Quality of life was measured using the validated Gastrointestinal Quality of Life Index (GIQLI) at baseline and after 6 weeks. With the GIQLI, scores range from 0 to 144, with higher scores representing better quality of life. RESULTS: Complete quality of life assessment was available in 29 patients (budesonide: n=17; placebo: n=12). At baseline, quality of life was low in patients with collagenous colitis (mean 76). After 6 weeks of treatment, the mean GIQLI score increased significantly in the budesonide group (from 67 to 92, p<0.001), but remained unchanged in the placebo group (86-88). The mean score of the dimensions symptoms (p=0.001), emotional functioning (p=0.003) and physical functioning (p=0.017) increased significantly in the budesonide group compared with the placebo group. A significantly larger proportion of patients in the budesonide group experienced improved stool consistency (p<0.01) and a significant reduction in the mean stool frequency compared with those in the placebo group (p<0.01). CONCLUSION: Quality of life is seriously reduced in patients with collagenous colitis. Six-week treatment with oral budesonide controlled-release capsules significantly improves quality of life and clinical symptoms compared with placebo in these patients. PMID- 15549327 TI - Adjuvant chemotherapy in curative resected colon carcinoma: 5 fluorouracil/leucovorin versus high-dose 5-fluorouracil 24-h infusion/leucovorin versus high-dose 5-fluorouracil 24-h infusion. AB - BACKGROUND: Adjuvant postoperative treatment with 5-fluorouracil (5-FU) and leucovorin in curatively resected stage III colon cancer significantly reduces the risk of cancer recurrence and improves survival. The impact of continuous 5 FU with and without leucovorin on survival and tumor recurrence was analyzed in this study compared with the effects of bolus 5-FU/leucovorin. PATIENTS AND METHODS: Patients with a curatively resected UICC stage III colon cancer were stratified according to T, N and G category and randomly assigned to receive one of the three adjuvant treatment schemes: 5-FU 450 mg/m2 and leucovorin 100 mg/m2 x 5 days every 4 weeks; six cycles, arm A; 24-h infusion of high-dose 5 FU/leucovorin 2,600 mg/m2 and 500 mg/m2, two cycles of six applications, arm B; 24-h infusion of high-dose 5-FU 2,600 mg/m2, two cycles of six applications, arm C. RESULTS: One hundred and forty-five patients enrolled into this study were eligible. To date, 28 patients have died; 9 on arm A, 11 on arm B, and 8 on arm C (P was nonsignificant). After a median follow-up time of 45 months, there was no statistical difference in survival and tumor recurrence between the three treatment arms. Adjuvant treatment in all arms was generally well tolerated; only a minority of patients experienced grade 3 and 4 toxicities. CONCLUSION: There is no statistical difference in efficacy and toxicity in patients receiving either high-dose 5-FU with or without leucovorin or the standard 5-FU bolus regime after a curative resection of a stage III colon cancer. PMID- 15549328 TI - Lycopene enhances UVA-induced DNA damage and expression of heme oxygenase-1 in cultured mouse embryo fibroblasts. AB - BACKGROUND: It has been suggested that carotenoids including lycopene may reduce the risk of photodamage. However, carotenoids are unstable under light exposure and may produce prooxidative effects under certain circumstances. AIM OF THE STUDY: We examined whether lycopene inhibits ultraviolet A (UVA)-induced DNA damage and the expression of heme oxygenase-1 (HO-1). We hypothesized that the breakdown of lycopene by UVA irradiation, rather than intact lycopene itself, causes oxidative damage. METHODS: Mouse fibroblasts, C3H10T1/2 (C3H), were first enriched with 10 microM of lycopene in the dark for 2 h before exposure to UVA (22.5 KJ/m2). Then, DNA damage measured by the single-cell gel electrophoretic assay (comet assay) and the expression of HO-1 measured by western blotting were determined. In addition, we exposed lycopene powder to UVA (22.5 KJ/m2) to prepare pre-irradiated lycopene (ILP). Then, C3H cells were incubated with ILP for 2 h, and DNA damage and the expression of HO-1 also were determined. RESULTS: We found that lycopene enrichment did not cause damage to DNA in C3H cells not irradiated with UVA. However, lycopene enrichment strongly induced DNA damage when cells were irradiated with UVA (by ca. 2-fold as compared to control). In addition, lycopene enhanced UVA-induced HO-1 expression by ca. 2.5-fold. UVA irradiation led to a significant loss of lycopene that had been pre-incorporated into C3H cells. When cells were incubated with lycopene that had been pre irradiated with UVA without subjecting the cells to further UVA irradiation, cellular DNA damage and expression of HO-1 were markedly increased, and these effects of irradiated lycopene were concentration-dependent. CONCLUSIONS: These results demonstrate that lycopene enhances UVA-induced oxidative stress in C3H cells, and they suggest that under UVA irradiation, lycopene may produce oxidative products that are responsible for the prooxidant effects. PMID- 15549329 TI - Hypoglycaemia causes degeneration of large myelinated nerve fibres in the vagus nerve of insulin-treated diabetic BB/Wor rats. AB - The aim of this study was to find out whether dysglycaemia causes neuropathy in the vagus nerve of insulin-treated diabetic BB/Wor rats. Specimens were collected from the left vagus nerve proximal and distal to the level of recurrent laryngeal branch and from the recurrent branch itself in control rats and diabetic BB/Wor rats subjected to hyper- or hypoglycaemia. Myelinated and unmyelinated axons were counted and myelinated axon diameters were measured by electron microscopy. In controls, the vagus nerve proximal to the recurrent branch exhibited three regions in terms of fibre composition: part A was mainly composed of large myelinated axons, part B contained small myelinated and unmyelinated axons, and part C contained mainly unmyelinated axons. The distal level resembled part C at the proximal level and the recurrent branch resembled parts A and B. In hyperglycaemic rats, a normal picture was found at the proximal and distal levels of the vagus nerve and in the recurrent branch. In hypoglycaemic rats, signs of past and ongoing degeneration and regeneration of large myelinated axons were found at the proximal and distal levels and in the recurrent branch. We conclude that hypoglycaemia elicits degenerative alterations in large myelinated axons in the vagus and recurrent laryngeal nerves in diabetic BB/Wor rats. The absence of signs of neuropathy in unmyelinated and small myelinated axons suggests that the sensory and autonomic components of the nerve are less affected. In contrast, the hyperglycaemic rats examined here did not show obvious degenerative alterations. PMID- 15549330 TI - Failure of caspase inhibition in the double-lesion rat model of striatonigral degeneration (multiple system atrophy). AB - In the present study we assessed the neuroprotective effects of the pan-caspase inhibitor z-VAD.fmk [N-benzyloxycarbony-valine-alanine-aspartate-(OMe) fluoromethylketone], and the caspase-3 inhibitor Ac-DEVD.CHO (acetyl-aspartate chloromethylketone) in the double-lesion rat model of striatonigral degeneration (SND), the core pathology underlying levodopa-unresponsive parkinsonism associated with multiple system atrophy (MSA). Male Wistar rats were divided into three groups, receiving either Ac-DEVD.CHO, z-VAD.fmk or normal saline before lesion surgery, comprising a sequential unilateral quinolinic acid (QA) lesion of the striatum followed by a 6-hydroxydopamine (6-OHDA) lesion of the ipsilateral medial forebrain bundle. At 2 weeks post lesion, all rats underwent testing of spontaneous nocturnal locomotor behavior in an automated Photobeam Activity System (FlexField). Immunohistochemistry was performed with tyrosine hydroxylase, dopamine and cyclic adenosine 3',5'-monophosphate-regulated phosphoprotein and glial fibrillary acidic protein antibodies. Morphometry was performed using computerized image analysis. Behavioral and morphological analysis failed to show striatal or nigral protection in caspase inhibitor-treated animals. Our findings suggest that anti-apoptotic strategies are unrewarding in the SND rat model and, therefore, alternative neuroprotective interventions such as anti-glutamatergic agents or inhibitors of microglial activation should be explored instead. PMID- 15549331 TI - Diapedesis of mononuclear cells across cerebral venules during experimental autoimmune encephalomyelitis leaves tight junctions intact. AB - Diapedesis of leukocytes across endothelial barriers is generally believed to require the opening of endothelial tight junctions. At the blood-brain barrier (BBB), endothelial cells are interconnected by complex tight junctions. Here, we show by serial section conventional electron microscopy that during experimental autoimmune encephalomyelitis mononuclear cells traverse cerebral microvessels by a transcellular pathway, leaving the endothelial tight junctions intact. Cerebral endothelial cells were found to form filopodia-like membrane protrusions on their luminal aspect, thus embracing the mononuclear cells and forming cup-like structures, and eventually pores, through which the traversing cell could reach the abluminal side. At the abluminal side endothelial cell protrusions surrounding a migrating inflammatory cell were found to be progressively lined with basal lamina, suggesting a change from luminal to abluminal membrane characteristics of endothelial cell membranes during inflammatory cell diapedesis. Morphological evidence for the involvement of tight junctions in the diapedesis of mononuclear cells across the inflamed BBB could not be obtained in any case. Taken together, the presence of morphologically intact tight junctions and our novel finding of the presence of a basal lamina on both sides of abluminal endothelial cell protrusions surrounding migrating inflammatory cells suggests that during experimental autoimmune encephalomyelitis diapedesis of mononuclear cells occurs via a transendothelial process. PMID- 15549332 TI - Ependymoma with neuropil-like islands: a case report with diagnostic and histogenetic implications. AB - We describe a case of ependymoma with neuronal differentiation in form of neuropil-like islands. A 6-year-old boy presented at clinical examination for a short history of headaches and vomiting. Brain computed tomography showed a large, partially cystic, parieto-occipital lesion. The tumor was composed by glial fibrillary acidic protein-positive round cells with a perivascular arrangement and scattered neuropil-like islands, showing intense positivity for synaptophysin. Despite radiotherapy, the tumor recurred, showing frank features of anaplasia, but lacking the neuropil-like islands. The histological features of the tumor are discussed in the light of the concept that neuronal differentiation can occur occasionally in gliomas of different lineage without affecting the expected biological behavior. PMID- 15549333 TI - Exclusive induction of tau2 epitope in microglia/macrophages in inflammatory lesions-tautwopathy distinct from degenerative tauopathies. AB - Tau2 antibody recognizes a phosphorylation-independent epitope that is pathologically modified as tau protein is phosphorylated to form neurofibrillary tangles of Alzheimer's disease (AD). Similar modification of tau2 epitope can be induced even in the absence phosphorylation of tau, as we first demonstrated in ischemic foci and in glial cytoplasmic inclusions (GCIs) of multiple system atrophy. This modification of tau2 epitope is distinguishable from those observed in degenerative tauopathies because (1) it is a conformational change, which is reversible upon exposure to a detergent; (2) it shows an absence of fibrils composed of phosphorylated tau protein; and (3) it is characterized by the lack of immunohistochemical labeling by anti-tau antibodies other than tau2. In this study, we expanded this observation to inflammatory foci of different pathologies (human immunodeficiency virus encephalopathy, progressive multifocal leukoencephalopathy or multiple sclerosis) by examining formalin-fixed, paraffin embedded sections immunostained with a panel of anti-tau antibodies. It was found that tau2 was the only anti-tau antibody that immunolabeled microglia/macrophages in these lesions, and this immunoreactivity was reversibly diminished upon exposure to a detergent. Exclusive apparition of tau2 immunoreactivity in these cells without neurofibrillary pathology may be a secondary event shared with ischemic foci and GCIs. It is, however, related to a unique conformational state of tau, possibly grouped under the name of "tautwopathy", that may represent an initial stage of tau deposition distinct from degenerative tauopathies characterized by fibrils composed of phosphorylated tau protein. PMID- 15549334 TI - Glycodelin A and differentiation of first trimester trophoblast cells in vitro. AB - AIM: The glycoprotein, glycodelin A (GdA) is a main product of the maternal decidua in the first trimester of pregnancy and is secreted into the amniotic fluid. The purpose of this study was to investigate the effect of GdA on secretion and surface markers of isolated first trimester trophoblasts in vitro. METHODS: Cytotrophoblasts were prepared from human first trimester placentae and incubated with varying concentrations of GdA or transfected separately with the expression plasmid of GdA. Supernatants were assayed for human chorionic gonadotropin (hCG) protein concentrations. Expression of human placental lactogen (hPL), mucin 1 (MUC1) and the Thomsen-Friedenreich (TF) epitope was analysed in stimulated trophoblast cells and in unstimulated controls by immunocytochemistry. RESULTS: Glycodelin A induced a reduced expression of hPL compared with unstimulated controls. Expression of MUC1 was not affected by GdA. Freshly isolated trophoblast cells showed no TF expression but became positive for this antigen after 96 h of cultivation. GdA-stimulated trophoblast cells inhibited TF expression after 96 h of cultivation. GdA plasmids induced a significantly higher hCG production in transfected cells than in cells transfected with the empty plasmid. CONCLUSIONS: The results obtained in this study suggest that GdA is involved in the differentiation of trophoblast cells. The treatment of GdA plasmid transfected trophoblast cells stimulated hCG production in isolated trophoblast cells and inhibited hPL and TF expression, suggesting a functional link between hCG and GdA. PMID- 15549335 TI - Study of abdominal circumference proportions in fetuses with growth disorders. AB - OBJECTIVE: Abdominal circumference is the ultrasound parameter that best correlates with fetal growth. The purpose of this study was to assess whether its proportions change in cases of growth disorders. METHODS: Using the abdominal protuberance ratio(APR) described in a previous paper, the abdominal circumference in fetuses affected by macrosomia and growth retardation was studied. RESULTS AND CONCLUSIONS: In both cases, a major tendency to a normal APR was observed, revealing that abdominal circumference in macrosomic and growth retarded fetuses changes as a whole, probably reflecting a global action of the factors participating in growth promotion or restriction. In cases of macrosomia, the minor slide to a high APR could reveal the specificity of the storage organs situated anteriorly in the abdomen. Otherwise, in cases of growth retardation, the slight tendency to a high APR would reveal a special reduction in the abdominal circumference in the posterior part of its surface, probably as a consequence of the flow redistribution mechanism. PMID- 15549336 TI - Trichinella spiralis and laryngeal carcinoma: a case report. AB - An uncommon presentation of Trichinella spiralis with squamous cell carcinoma of the larynx is reported. The possible carcinogenic potential of trichinosis and its management are discussed. PMID- 15549337 TI - Rare case of pyogenic granuloma of the tongue in an 8-week-old infant. AB - We present a case of a large pyogenic granuloma of the tongue causing swallowing difficulties in an 8-week-old infant. In the literature, we found no case reports of pyogenic granuloma in such a young infant. It is a quickly growing, hypertrophic inflammatory change consisting of exceptionally richly vascularized granular tissue. The granuloma was located in the central part of the tongue and appeared dark pink in color. It was an easily bleeding pedunculated tumor measuring 3 x 2.5 cm. The etiology of the tumor is unknown; it can arise in sites of previous injury, chronic irritation and infection. The course of the disease is benign and painless, though it can be recurrent. No recurrence was found in the control examination after 3 years. PMID- 15549338 TI - Laryngeal chondroma presenting as a laryngopyocele. AB - The abnormal dilatation of saccules in direct communication with the laryngeal lumen is termed laryngocele. A simple laryngocele is an air-filled dilatation of the saccule and is mostly asymptomatic. When the neck of the laryngocele is obstructed, the laryngocele becomes filled with mucus. If a mucus-filled laryngocele is infected, it is called a laryngopyocele. The etiology of laryngoceles is not well understood. However, there is an association between laryngoceles and carcinoma of the supraglottic larynx. The main reasons are probably obstruction and mucosal retention of the saccule, increasing intralaryngeal pressure due to airway obstruction or abnormal contraction of the involved laryngeal muscles due to tumoral spreading. We present a patient with laryngopyocele that arose because of a laryngeal chondroma. PMID- 15549339 TI - A simple technique for nasogastric feeding tube insertion. AB - Nasogastric tubes are increasingly used in the management of a diverse group of patients who generally require short-term enteral feeding. Various techniques of insertion have been described emphasizing the fact that as yet there is no simple and safe method. Nasogastric intubation in head and neck cancer patients may be especially difficult following radiotherapy due to difficulties in swallowing secondary to edema, mucositis, abnormal anatomy and altered sensation. In this paper, we describe a simple technique that evolved from experience of passing enteral feeding tubes in head and neck cancer patients. The feeding tube is inserted through the appropriate nasal cavity, and at 21 cm (8 inches) from the anterior nares in the average adult (corresponding to a few millimeters above the arytenoids), the patient is asked to vocalize by saying 'eeeee' in a high pitched tone. The tube is then advanced into the esophagus while the patient is vocalizing. This technique has been successfully carried out in 22 consecutive patients, thereby avoiding the use of more invasive methods. PMID- 15549340 TI - Changes in the gene expression pattern of cytokeratins in human respiratory epithelial cells during culture. AB - The replacement of extensive tracheal defects resulting from intensive care medicine, trauma or large resections is still challenged by the re epithelialization of an autologous or alloplastic trachea replacement. Therefore, this study was performed to investigate the potential of culture-expanded human respiratory epithelial cells (hREC) to regenerate a functional epithelium for tracheal tissue engineering. hREC from seven male nasal turbinates were freshly isolated, expanded on a collagenous matrix and subsequently cultured in high density multi-layers to allow epithelial differentiation. The composition of epithelial cells in native respiratory epithelial tissue and culture-expanded hREC was analyzed by histological staining with Alcian blue and by immunohistochemical staining of cytokeratin pairs CK1/10 and CK5/14 with the antibodies 34betaE12 and CD44v6. Differentiation of culture-expanded hREC was further characterized by gene expression analysis of cytokeratins CK5, CK13, CK14 and CK18 using semi-quantitative real-time RT-PCR technique. Histological and immunohistochemical staining of culture-expanded hREC demonstrated basal cells covering the collagenous matrix. These cells formed a cellular multi-layer, which was composed of a basal layer of undifferentiated basal cells and an upper layer of cells differentiating along the squamous metaplasia and ciliated cell lineage. Lineage development of culture-expanded hREC was further documented by the induction of cytokeratins CK13 and CK18. Our results suggest that culture expanded hREC have the potential to colonize collagen-coated biomaterials and to regenerate epithelial cell types for tracheal tissue engineering. PMID- 15549341 TI - Actinomycosis of the middle turbinate: an unusual cause of nasal obstruction. AB - Actinomycosis is a rare chronic infectious disease caused by Actinomyces israelii, which is an anaerobic filamentous, gram-positive saprophyte organism of the oral cavity. Historically, these bacteria were known as fungi because of their light microscopic appearance. Actinomycosis consists of three different forms: cervico-facial (the most common form), abdominal and pulmonothoracic. It commonly involves the head and neck region including the mandible, paranasal sinuses, lacrimal gland, parotid gland and orbit. Poor oral hygiene and dental diseases have been known to be the source of actinomycosis. Actinomycosis is diagnosed with positive culture or detecting actinomyces colonies and sulfur granules in histopathologic specimens. The treatment of choice is surgical excision of the lesion and long-term penicillin therapy. Actinomycosis of the internal nose is extremely rare. There was only one nasal septum actinomycosis reported in the English literature, but there was no lateral nasal wall actiomycosis regarding the turbinate. Therefore, actinomycosis should not be overlooked for the differential diagnosis of intra-nasal lesions for the initiation of appropriate and early treatment. PMID- 15549342 TI - Burden of relatives and predictors of burden. Baseline results from the Munich 5 year-follow-up study on relatives of first hospitalized patients with schizophrenia or depression. AB - In the present study, part of the Munich 5-year follow-up study on key relatives of first-hospitalized schizophrenic and depressive patients, baseline results with respect to relatives' burden and predictors of burden are presented. Basing on a transactional stress model the following hypothesis was tested: the impact of the patients' illness on their relatives' stress outcome is moderated by the psychosocial resources of the relatives. Stress outcome was measured in terms of objective and subjective burden, well-being, self-rated symptoms and global satisfaction with life. Potential moderating variables included age and gender, generalized stress response and illness-related coping strategies, beliefs of control,perceived social support,personality factors, expressed emotion and life stressors. A total of 83 relatives, whose ill family members had been hospitalized in the Department of Psychiatry of the Ludwig-Maximilians-University of Munich for the first time, participated in the study. Findings did not entirely support the hypothesis. On the one hand, relatives' stress outcome was independent of the objective stressors (severity of the illness, kind of symptoms, level of psychosocial functioning at admission). On the other hand, burden was significantly associated with several psychosocial resources and dispositions of the relatives. Multivariate linear regression analyses indicated that expressed emotion, emotion-focused coping strategies and generalized negative stress response are the most relevant predictors of burden. It is argued that a multidimensional approach in burden assessment is necessary and has relevant implications for improving family intervention strategies. PMID- 15549343 TI - Suicide attempts and impulsivity. AB - Impulsivity in suicidal behavior can describe the attempt (state) or the attempter (trait). There are no studies simultaneously measuring attempt impulsivity and attempter impulsivity in representative samples of suicide attempts. A one-year study of 278 suicide attempts in a general hospital tested the continuous versus dichotomous relationship between attempter impulsivity (Barratt Impulsiveness Scale) and attempt impulsivity (low scores in the planning subscale of Beck's Suicidal Intent Scale). Attempter impulsivity was not a good predictor of attempt impulsivity independently of how both dimensions were measured (continuous or dichotomous ways). Impulsive attempts were associated with low lethality and lack of depression. Opportunities for prevention of suicide attempts in major depression and some personality traits may exist but require attentive monitoring of suicidal ideation and intent. PMID- 15549344 TI - Clinical effects of COX-2 inhibitors on cognition in schizophrenia. AB - An activation of pro-inflammatory cytokines in the central nervous system is associated with cognitive disturbances. This process is mediated by prostaglandins and cyclo-oxygenase-2 (COX-2). COX-2 inhibitors have been suggested to show beneficial effects in disorders associated with cognitive disturbance, although clinical effects on cognition have not been shown until today. Data from a schizophrenia study were reevaluated under the aspect whether an effect on the positive and negative syndrome scale (PANSS) factor cognition can be observed during therapy with the COX-2 inhibitor celecoxib add on to risperidone in comparison to riperidone alone. Beside the effect on the PANSS total score, the effect on the cognition factor was the most pronounced using the analysis of covariance compared to all other factors of the PANSS (p < 0.06). Although suggestions of basic research led to an expected therapeutic effect of COX-2 inhibitors on cognition, this effect could not yet be shown clinically. In schizophrenia, the effect on cognition contributes to the therapeutic effect of COX-2 inhibitors. PMID- 15549345 TI - How representative of everyday clinical populations are schizophrenia patients enrolled in clinical trials? AB - INTRODUCTION: There has been considerable discussion whether clinical trials accurately depict everyday practice. Restrictive inclusion/exclusion criteria, ethical considerations, differences in the severity of psychopathology between clinical and trial patients, or safety issues may bias results, which in turn may rather represent outcome for the "ideal" than for the "average"patient. Therefore, translation into psychiatric practice may be difficult. METHODS: A retrospective case-control study was performed. Schizophrenia inpatients at the LMU Department of Psychiatry, Munich, Germany, who had participated in clinical trials were compared to regular patients serving as controls. Probands and controls were matched by DSM-IV diagnosis, gender and age. The AMDP module, CGI and GAF were used to compare psychopathology. In addition, charts were reviewed for medication dosages, concurrent medical and neurological illness, and clinical history such as age of onset or family history. RESULTS: A total of 200 probands (100/100) were enrolled in the study. With respect to psychopathology, formally thought disordered or suicidal patients were significantly less likely to be study participants (n = 3) than controls (n = 22; p < or = 0.05). Similarly, negative schizophrenia symptoms were significantly less often present in study participants (n = 17) than in controls (n = 38; p < or = 0.05). Study participants were also medically and neurologically healthier than controls. (p = 0.05 respectively). No differences in overall illness severity as depicted by CGI and GAF were observed. CONCLUSION: We found the patients included in our clinical trials representative of the patient encountered in routine clinical practice. Adherence to inclusion and exclusion criteria prevents inclusion of severely ill (e. g. suicidal) patients requiring a more intensive treatment setting. Illness severity was found to be similar in trial participants and controls, and indicates an overall comparably severe psychopathology. The more chronic, rather treatment refractory patients were also not reflected in the trial participant pool; this population may arguably not represent the average clinical patient either. A more careful administration of antipsychotic medication was found in trial participants and may effectively be considered "good clinical practice". PMID- 15549346 TI - Detection of alcohol consumption in suicides. AB - Screening instruments for detection of alcohol consumption, abuse, and dependence for use in psychological autopsy studies with case control design are not validated. Therefore, interrater and test-retest reliability of the Luebeck Alcohol Dependence and Abuse Screening Test (LAST) and the usability of this test for the psychological autopsy method were investigated. Alcohol consumption was evaluated by a semi-structured interview including the Structured Clinical Interview for DSM-IV Axis I (SCID-I) and the LAST in 163 completed suicides (mean age 49.6 +/- 19.3 years; 64.4% men) and by personal interview in 396 population based controls (mean age 51.6 +/- 17.0 years; 55.8% men). Of the controls, 35 were additionally assessed by interviewing informants; these results were compared with those generated by personal interview. Comparison of LAST scores by personal and informant's interview of controls generated a Spearman correlation coefficient of 0.74 (P < 0.0001). The LAST (7 item-version, cut-off of 2) revealed high sensitivity and specificity for alcohol abuse and dependence, in both controls and suicides. LAST scores were significantly associated with high, frequent, and hazardous alcohol consumption (P < 0.001) in suicides. Our findings provide support for reliability and validity of identifying individuals with alcohol dependence and abuse obtained through the best-estimate method using the LAST. This 7-item questionnaire can be recommended as a useful tool for the psychological autopsy procedure in postmortem research. PMID- 15549348 TI - On various possibilities in pulsed radiation biochemistry and chemistry. AB - Several experiments are described that relate to the application of new regimes of radiation action on enzymes in vitro and some other materials. These regimes have recently come into practice due to the appearance of a new generation of devices with very short high-energy pulses of ionizing radiation. It is shown that the term "flash radiation biochemistry" in its perfect sense has to be used at the condition of the overlapping individual effective interaction microvolumes (e.g. spurs and blobs) realized during a time interval (radiation pulse duration) that is low compared with the corresponding physical-chemical process. In this situation a number of unexpected effects occur at very low absolute doses. These processes are analyzed in terms of their non-stationary and non-diffusive developments. PMID- 15549352 TI - Disease-modifying therapy in MS: a critical review of the literature. Part I: Analysis of clinical trial errors. AB - Several types of errors are commonly made during the conduct, analysis, and interpretation of clinical trials in multiple sclerosis (MS). These include statistical errors of overestimating the significance of trial results, particularly when multiple endpoints are evaluated. They also include errors arising from the use of inappropriate covariate analyses, meta-analyses, and post hoc subgroup analyses. Interpretation of trial results can also be confounded by regression to the mean, by post hoc data re-analysis, and by the use of a non concurrent control population. As these kinds of errors continue to plague the medical literature, it has become important for physicians to be able to assess critically the reports of clinical trial results. In turn, this has made it necessary for physicians to become familiar with the rudiments of the fields of statistics, epidemiology, and trial design. It is the purpose of this manuscript, therefore, to provide an overview of these principles through a detailed analysis of these kinds of clinical trial errors, together with examples that have actually occurred in the recently published MS literature. PMID- 15549350 TI - What do we know about the mechanism of action of disease-modifying treatments in MS? AB - Multiple sclerosis (MS), a chronic inflammatory disorder of the central nervous system (CNS), 2 results in damage to axons and their surrounding myelin sheath. The exact cause of inflammation remains unclear, but an autoimmune response directed against CNS antigens is suspected. MS can affect the brain, optic nerve and spinal cord, thus causing many neurological symptoms. These can include limb numbness or weakness, sensory or motor changes, ataxia, blurry vision, painful eye movements, bladder and bowel dysfunction, decreased memory, fatigue and effective disorders. This article will include a concise overview of the pathogenesis of MS in order to set the stage for subsequent discussion of the mechanisms of action of disease-modifying treatments, and whether these should influence our treatment choices. Although the exact pathogenesis of MS is not fully understood, current knowledge has already led to the development of effective treatments, namely interferon (IFN) 3 and glatiramer acetate, both of which have been shown to reduce relapse rates, while IFN 3- 1 a also reduces confirmed disability progression. Further increases in our understanding of the pathogenesis of MS are likely to assist in the identification of new targets for disease-modifying therapies and in the optimisation of current treatments.. PMID- 15549353 TI - Optimising MS disease-modifying therapies: antibodies in perspective. AB - A proportion of people with multiple sclerosis (MS) treated with interferon (IFN) a develop neutralising anti-IFN beta antibodies (NABs). The immunogenicity of the available commercial compounds relates to the genetic structure of the IFN beta molecule, its mode of production, glycosylation status, aggregate formation, commercial formulation, potency, dose, frequency and, possibly, route of administration. At present, it is not possible to predict who will develop NABs usually appear within the first 2 years of starting therapy. In patients treated with IFN beta in whom NABs persist for a significant period of time, their presence is associated with a reduction in both the biological effects and clinical efficacy. Approximately one third of NAB-positive patients with a titre > 20 NU/mL will revert to NAB-negative status with long-term follow-up. The persistence of NABs appears to be linked to the type of IFN beta treatment as well as the titre of antibodies. The overall efficacy of IFN beta and, hence, of any biological disease-modifying treatment (DMT) would be substantially improved if the development of NABs could be prevented or reversed. Although the overall efficacy of IFN beta in MS is relatively modest, the efficacy in individuals who remain NAB-negative is considerably better than in those who become persistently NAB-positive. One could argue that when comparing the 'true' clinical efficacy of different IFN beta products, the comparisons should be limited to the cohorts that remain NAB-negative. As a corollary, the therapeutic efficacy of IFN beta could be maximised if patients who tolerate higher-dose preparations could be prevented from developing persistent NABs. Strategies employed to prevent or reverse the development of NABs with other biological compounds (e. g. insulin, factor VIII, IFN beta, recombinant human erythropoietin) include improvements in the manufacturing process, immunosuppression, induction of tolerance and deimmunisation, and these should be considered in relation to biological DMT therapy as part of future clinical studies. PMID- 15549354 TI - MRI: role in optimising treatment. AB - Magnetic resonance imaging (MRI) is an extremely sensitive diagnostic tool that provides us with highly detailed images of the living human brain. Since it was first applied to multiple sclerosis (MS) in clinical trials in the 1980s, MRI has become established as a reliable, sensitive and reproducible technique for studying the pathophysiology of this complex disease. It has provided a variety of surrogate measures for clinical trials, and continues to offer new methods for the detection and monitoring of the physical and chemical changes in the brains of living patients. The unique sensitivity of conventional MRI measures for detecting MS pathology has made MRI an attractive tool for optimising treatment in clinical practice for individual patients. PMID- 15549355 TI - Importance of benefit-to-risk assessment for disease-modifying drugs used to treat MS. AB - Interferon (IFN) beta has been shown to be an effective therapy in pivotal studies of multiple sclerosis (MS), with differences in outcomes based on dose and/or frequency of administration. Glatiramer acetate (GA) has also shown to have an effect on relapses and magnetic resonance imaging measures, but not on disability. All products are associated with adverse events, and utilisation of a specific therapy needs to consider benefit in relation to risk. Evidence-based medicine provides a means of assessing benefit and risk in the context of the number of patients one needs to treat to obtain benefit (NNT) compared with the number needed to treat for an adverse outcome (NNH). Efficacy and safety data are presented from IFN beta-1a (Rebif) clinical trials, including relevant NNT and NNH values, to allow assessment of the overall benefit-to-risk ratio compared with placebo. Additional comparisons are made with published data for other IFN products and GA. The indirect comparative information reviewed demonstrates that IFN appears to have a better benefit- to-risk ratio than GA. Indirect comparisons suggest better efficacy of thrice weekly (tiw) IFN beta-1a compared with alternate-day IFN beta-1b, but no direct comparative data are available. Direct comparative data show that IFN beta-1a at a dose of 44 mcg tiw has a favourable benefit-to-risk ratio compared with both 22 mcg tiw and 30 mcg once weekly, suggesting that 44 mcg tiw currently has the best benefit- to-risk ratio for the treatment of relapsing MS. PMID- 15549356 TI - Disease-modifying therapy in MS: a critical review of the literature. Part II: Assessing efficacy and dose-response. AB - The use of so-called evidence-based medicine represents a structured way in which to critically assess the medical literature with the goal of defining the value of different therapeutic interventions and, ultimately, improving both physician decision-making and patient outcome. This is not a consensus-based process of the type that has often been employed previously in the development of treatment guidelines. Rather, these assessments involve a series of structured steps. Initially, the specific clinical questions to be answered are defined and the evidence is assembled following a structured literature search. Then, the individual studies are classified as to the quality of the evidence provided, and, finally, using a set of pre-specified rules, this evidence is translated into specific recommendations and conclusions. In this manner, evidence-based medicine can be a very powerful tool for practicing physicians and, consequently, it is important that they become familiar with the fundamentals of this analytical approach. It is the purpose of this manuscript, therefore, to provide an overview of this process using examples from two recently completed assessments on disease-modifying therapies in multiple sclerosis. PMID- 15549357 TI - Alternatives to current disease-modifying treatment in MS: what do we need and what can we expect in the future? AB - Disease-modifying treatments (DMTs) for multiple sclerosis (MS) are now widely available, and their beneficial effects on relapse rates, magnetic resonance imaging outcomes and, in some cases, relapse-related disability have been shown in numerous clinical studies. However, as these treatments are only partially effective in halting the MS disease process, the search for improved treatment regimens and novel therapies must continue. Strategies to improve our therapeutic armamentarium have to take into account the different phases or parts of the pathogenesis of the disease. Available treatments address systemic immune dysfunction, blood-brain barrier permeability and the inflammatory process in the central nervous system. Currently, patients who fail to respond adequately to first-line DMTs are often considered as candidates for intensive immunosuppression with cytostatic agents or even autologous stem cell transplantation. However, new approaches are being developed. Combination therapies offer an alternative approach that may have considerable potential to improve therapeutic yield and, although likely to present considerable challenges in terms of trial design, this certainly seems to be a logical step forward in view of the complex pathology of MS. Several new drugs are also being developed with the aim of providing more effective, convenient and/or specific modulation of the inflammatory component of the disease. These treatments include humanised monoclonal antibodies such as the anti-VLA-4 antibody natalizumab, inhibitors of intracellular activation, signalling pathways and T-cell proliferation, and oral immunomodulators such as sirolimus, teriflunomide or statins. There remains, however, an urgent need for treatments that protect against demyelination and axonal loss, or promote remyelination/regeneration. Due to the chronicity of MS, the therapeutic window for neuroprotective agents is wider than that following stroke or acute spinal cord injury, and may therefore allow the use of some drugs that have proven disappointing in other situations. Novel potential neuroprotective agents such as alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid antagonists and ion-channel blockers will be entering Phase II trials in MS in the near future, and it is hoped that these agents will mark the start of a new era for DMTs for MS. PMID- 15549358 TI - Predicting the course of relapsing-remitting MS using longitudinal disability curves. AB - BACKGROUND AND OBJECTIVE: Multiple sclerosis (MS) is a chronic, progressive disease of the central nervous system that generally occurs in adults under the age of 40 years and ultimately leads to severe neurological disability. Following the progression of MS by monitoring changes in disability levels can facilitate treatment decisions taken by physicians. The aim of this review is to present longitudinal disability curves enabling the assessment of disease progression in patients with relapsing-remitting (RR)MS. METHODS: Patients with a definite diagnosis of MS and an RR disease course were identified using the Multiple Sclerosis Center computerised database. Patients were stratified into major percentile groups based on their Expanded Disability Status Scale (EDSS) score 1 year after disease onset. Model disability curves for each percentile were constructed using mean consecutive EDSS scores for 10 years after disease onset. Model curves were generated by smoothing (parametric and non-parametric regression) and curve approximation (linear regression and moving averages). The predictive ability of model curves was validated by superimposing data from a separate group of patients with RRMS. RESULTS: Disability curves were constructed using data from 1001 patients. A significant difference between the initial percentile assignment and disability progression was indicated by the log-rank test (p < 0.001). Kaplan-Meier and life table analyses demonstrated the validity of the model in predicting disease progression. The probability of experiencing more severe disability than predicted (i. e. deviating from the initial percentile to a higher percentile over time) ranged from 6.5 % (50th percentile) to 15.4 % (75th percentile), while the probability of experiencing less severe disability than predicted (i. e. deviating from the initially assigned percentile to a lower percentile over time) ranged from 6.9% (50th percentile) to 1.6 % (75th percentile). Both suggest reasonable predictive validity. CONCLUSION: In MS, longitudinal disability curves can help to assess individual patient disability, map the effects of immunomodulatory treatments over time, and generally build on the overall clinical impression of disease progression. Such models can act as a tool to aid and support the clinical decision-making process. This review is based on the study published in Multiple Sclerosis (2003) 9:486 491. PMID- 15549359 TI - Improving MS patient care. AB - Multiple sclerosis (MS) is a multidimensional, chronic, neurological disease affecting young people that often interferes with their life and career plans. Patient care begins at the time of diagnosis, and particular emphasis should be placed on proper education about the variable disease course and treatment options. Clinical assessment at regular intervals using quantitative measures is recommended in order to obtain important information on the effects of disease modifying and/or symptomatic treatment. In addition to optimising therapy, it is imperative to develop effective and properly resourced care services for MS patients that will address their complex and lifelong requirements. PMID- 15549360 TI - MS patient management: the Analog Model. AB - The Analog Model was developed for use in patients with multiple sclerosis (MS) at an international roundtable meeting in 2001. Three key disease components are monitored, namely relapses, disability progression and magnetic resonance imaging activity. Its use provides a rapid, straightforward means of assessing the efficacy of disease-modifying therapy and the need for treatment change. The Analog Model thus assists in optimising the management of patients with MS. PMID- 15549361 TI - Can databasing optimise patient care? AB - Long-term, prospective databasing of multiple sclerosis (MS) information provides a useful resource for natural history studies. Furthermore, it is the only way to address the question of whether early treatment eliminates or delays the inevitable and irreversible clinical worsening that is the hallmark of the late phase of the illness. Due to the variable nature of MS, it is useful to monitor large numbers of individuals over time. The limitations of single databases may be overcome by regional, national or international pooling of data. In this paper, the Italian Multiple Sclerosis Database Network (MSDN) and the international web-based MSBase registry are described. PMID- 15549363 TI - Treatment optimisation in MS: progress to date and goals for the future. PMID- 15549365 TI - Expression and amplification of therapeutic target genes in retinoblastoma. AB - PURPOSE: We set out to evaluate alterations of the therapeutic target genes KIT (CD 117), EGFR, and HER-2 in human retinoblastoma. METHODS: Ninety-five formalin fixed, paraffin-embedded retinoblastomas were brought into a tissue microarray (TMA) format. Immunohistochemistry was performed to analyze the expression of CD117, EGFR, and HER-2. Fluorescence in situ hybridization (FISH) was utilized for detection of EGFR amplifications. Three tumors with strong CD117 positivity were sequenced for KIT exon 11 mutations. RESULTS: Detectable CD117 expression was seen in 19% of all interpretable cases. Sequence analysis of the three tumors with the strongest CD117 expression revealed no mutations. EGFR was positive in 14% of all cases. No EGFR amplification was observed by FISH, however. All tumors were negative for HER-2 expression. CONCLUSIONS: Our data suggest that selected cases of retinoblastoma may be candidates for anti-EGFR and imatinib mesylate (STI571) therapy. PMID- 15549366 TI - Changes in P2X3 purinoceptors in sensory ganglia of the mouse during embryonic and postnatal development. AB - The expression of the P2X(3) nucleotide receptor in embryonic day 14-18, postnatal day 1-14 and adult mouse sensory ganglia was examined using immunohistochemistry. Nearly all sensory neurons in dorsal root ganglia, trigeminal ganglia and nodose ganglia in embryos at embryonic day 14 expressed P2X(3) receptors, but after birth there was a gradual decline to about 50% of neurons showing positive immunostaining for P2X(3). In embryos there were only small neurons, while from postnatal day 7 both large and small neurons were present. Isolectin B(4) (IB(4))-positive neurons in dorsal, trigeminal and nodose ganglia did not appear until birth, but the numbers increased to about 50% by postnatal day 14 when a high proportion of IB(4)-positive neurons were also positively labelled for the P2X(3) receptor. About 10% of neurons in dorsal, trigeminal and nodose ganglia were positive for calcitonin gene-related peptide in embryos, nearly all of which stained for P2X(3) receptors. This increased postnatally to about 35-40% in adults, although only a few colocalised with P2X(3) receptors. Neurofilament 200 was expressed in about 50% of neurons in trigeminal ganglia in the embryo, and this level persisted postnatally. All neurofilament 200-positive neurons stained for P2X(3) in embryonic dorsal root ganglia, trigeminal ganglia and nodose ganglia, but by adulthood this was significantly reduced. The neurons that were positive for calbindin in embryonic dorsal, trigeminal and nodose ganglia showed colocalisation with P2X(3) receptors, but few showed colocalisation postnatally. PMID- 15549367 TI - Exercise capacity in young adults with hypertension and systolic blood pressure difference between right arm and leg after repair of coarctation of the aorta. AB - Coarctation of the aorta represents 5-7% of congenital heart defects. Symptoms and prognosis depend on the degree of stenosis, age at surgery, surgical method and the presence of other heart defects. Postoperative complications are hypertension, restenosis and an abnormal blood pressure response during exercise. This study includes 41 patients, 15-40 years old, operated in the period 1975 1996. All were exercised on a treadmill until maximal oxygen consumption was achieved. Blood pressure was measured in the right arm and leg before and immediately after exercise, and in the right arm during exercise. Oxygen consumption was monitored and we defined an aerobic phase, an isocapnic buffering phase and a hypocapnic hyperventilation phase. The resting systolic blood pressure correlates with the resting systolic blood pressure difference between right arm and leg. A resting systolic blood pressure difference between the right arm and leg of 0.13 kPa (1 mmHg) to 2.67 kPa (20 mmHg) corresponds with a slight increase in resting systolic blood pressure. This rise in blood pressure increases the aerobic phase of the exercise test, helping the patients to achieve higher maximal oxygen consumption. A resting systolic blood pressure difference of more than 2.67 kPa (20 mmHg) corresponds with severe hypertension and causes reduction in the aerobic phase and maximal oxygen consumption. Resting systolic blood pressure and resting systolic blood pressure difference between the right arm and leg are not indicators for blood pressure response during exercise. Exercise testing is important to reveal exercise-induced hypertension and to monitor changes in transition from aerobic to anaerobic exercise and limitation to exercise capacity. PMID- 15549368 TI - Effects of active recovery between series on performance during an intermittent exercise model in young endurance athletes. AB - The purpose of our study was to compare time to exhaustion ( t(lim)) and time spent at a high level of oxygen uptake (V(.)O(2)) during two high-intensity short intermittent exercises (30 s-30 s) realized with or without series. Eleven young endurance-trained athletes [16.6 (0.4) years] took part in three field tests until exhaustion: (1) a maximal graded test to measure their maximal aerobic velocity (MAV) and maximal oxygen uptake (V(.)O(2max)); (2) and (3) two randomized intermittent exercises (30 s at 110% of MAV alternated with 30 s at 50% of MAV): one alternating repetitions non-stop (IE) and another including 4 min recovery every six repetitions (IEs). The mean t(lim) measured during IEs was significantly longer than IE [respectively 960.0 (102.0) s vs 621.8 (56.2) s]. The time spent at V(.)O(2max)( t(V(.)O2max)) and the time spent above 90% of V(.)O(2max)( t(90%V(.)O2max)) did not differ significantly according to the type of exercise: with or without series [respectively t(V(.)O2max) was 158.2 (59.7) s vs 178.0 (56.5) s and t(90%O2max) was 290.4 (84.3) s vs 345.0 (61.6) s] but when expressed as a relative value, t(90%O2max) during IEs was significantly lower than during IE [respectively 36.4 (10.4)% t(lim) vs 58.3 (8.7)% t(lim)]. Despite a significant decrease ( P<0.005) of time to achieve 90% of V(.)O(2max) at the start of each series during IEs [respectively 165.0 (43.1) s for the first series and 82.5 (15.8) s for the second series ( n=6)] the time spent under 90% of V(.)O(2max) limited the t(90%V(.)O2max) during each series. In conclusion, our results showed that intermittent exercise with series does not permit an increase in the time spent at a high level of V(.)O(2); however, the athletes performed more repetitions of short intense exercise. PMID- 15549369 TI - Effects of hyperthermia on the metabolic responses to repeated high-intensity exercise. AB - In this study, we investigated the metabolic and performance responses to hyperthermia during high-intensity exercise. Seven males completed two 30-s cycle sprints (SpI and SpII) at an environmental temperature of 20.6 (0.3) degrees C [mean (SD)] with 4 min recovery between sprints. A hot or control treatment preceded the sprint exercise. For the hot trial, subjects were immersed up to the neck in hot water [43 degrees C for 16.0 (3.2) min] prior to entering an environmental chamber [44.2 (0.8) degrees C for 30.7 (7.1) min]. For the control trial, subjects were seated in an empty bath (15 min) and thereafter in a normal environment [20.2 (0.6) degrees C for 29.0 (1.9) min]. Subjects' core temperature prior to exercise was 38.1 (0.3) degrees C in the hot trial and 37.1 (0.3) degrees C in the control trial. Mean power output (MPO) was significantly higher in the hot condition for SpI [683 (130) W hot vs 646 (119) W control ( P<0.025)]. Peak power output (PPO) tended to be higher in the hot trial compared with the control trial for SpI [1057 (260) W hot vs 990 (245) W control ( P=0.03, NS)]. These differences in power output were a consequence of a faster pedal cadence in the hot trial ( P<0.025). There were no differences in sprint performance in SpII in the hot trial compared to the control trial; however, MPO was significantly reduced from SpI to SpII in the hot condition but not in the control condition ( P<0.025). Plasma ammonia was higher in the hot trial at 2 min post-SpI [169 (65) micromol l(-1 )hot vs 70 (26) micromol l(-1) control ( P<0.01)], immediately and at 2 min post-SpII [231 (76) micromol l(-1) hot vs 147 (72) micromol l(-1) control ( P<0.01)]. Blood lactate was higher in the hot trial compared with the control trial at 5 min post-SpII ( P<0.025). The results of this study suggest that an elevation in core body temperature by 1 degrees C can improve performance during an initial bout of high-intensity cycle exercise but has no further beneficial effect on subsequent power production following a 4-min recovery period. PMID- 15549370 TI - Effects of lumbar extensor fatigue and fatigue rate on postural sway. AB - Falls from heights resulting from a loss of balance are a major concern in the occupational setting. Previous studies have documented a deleterious effect of lower extremity fatigue on balance. The purpose of this study was to investigate the effect of lumbar extensor fatigue on balance during quiet standing. Additionally, the effects of fatigue rate on balance and balance recovery rate were assessed. Eight center-of-pressure-based measures of postural sway were collected from 13 participants, both before and after a protocol that fatigued the lumbar extensors to 60% of their unfatigued maximum voluntary exertion force. In addition, postural sway was measured for 30 min after the fatiguing protocol, at 5-min intervals, to quantify balance recovery rate during recovery from fatigue. Two different fatigue rates were achieved by fatiguing participants over either 10 min or 90 min. Results show an increase up to 58% in time-domain postural sway measures with lumbar extensor fatigue, but no change in frequency domain measures. Fatigue rate did not affect the magnitude of these postural sway increases, nor did it affect the rate of balance recovery following fatigue. Statistical power for the latter result, however, was low. These results show that lumbar extensor fatigue increases postural sway and may contribute to fall from-height accidents. PMID- 15549371 TI - The influence of training level and surgical experience on survival in colorectal cancer. AB - BACKGROUND AND AIMS: The effect of surgical training level, experience, and operation volume on complications and survival in colorectal cancer during a 10 year period in a medium-volume university hospital was retrospectively studied. PATIENTS AND METHODS: Four hundred and fifty-six patients were resected for primary colorectal adenocarcinoma during the 10-year period of 1981-1990, and of these, 387 patients underwent resection with curative intent. The surgeons were divided into three groups according to training level and volume: group 1, surgeons in training and other surgeons operating annually on only 1-4 patients; group 2, surgeons specializing in gastrointestinal surgery (average annual volume 4-13 operations); group 3, specialists in gastrointestinal surgery (average annual volume 3-8 operations). Postoperative morbidity and mortality rates, as well as long-term survival rates, were analysed, and comparisons were made between the patients in the three groups. RESULTS: There were no statistically significant differences between the three groups in postoperative morbidity or mortality. Cancer-specific 5-year survival rate of all patients was 57%, and that of those resected in the aforementioned three groups was 51%, 63%, and 55%, respectively, P=0.087. The 5-year survival rates for colon cancer were 59% (total), 52%, 69%, and 58%, respectively, P=0.067, and for rectal cancer were 51% (total), 42%, 53%, and 52%, respectively, P=0.585. CONCLUSION: There were no significant differences in the rates of postoperative mortality, morbidity, and long-term overall survival between the volume groups. However, in patients with colon cancer, there was a trend for better survival for those operated on by the surgeons specializing in gastrointestinal surgery, and in rectal cancer patients, a tendency of fewer local recurrences in those operated on by the specialist surgeons. PMID- 15549373 TI - Generation and properties of ascorbic acid-overproducing transgenic tobacco cells expressing sense RNA for l-galactono-1,4-lactone dehydrogenase. AB - L-Galactono-1,4-lactone dehydrogenase (GalLDH; EC 1.3.2.3) is the last enzyme in the putative L-ascorbic acid (AsA) biosynthetic pathway of plants. Here, we show for the first time that the overexpression of GalLDH can increase AsA content in tobacco (Nicotiana tabacum L.) BY-2 cells. To see the effect, we analyzed the properties of these AsA-overproducing transgenic cell lines, especially in relation to AsA content of cells, cell division, senescence and resistance to oxidative stress. The mitotic index in AsA-overproducing cells was higher than in wild-type cells. Moreover, the browning of these cells was markedly restrained, and the proportion of dead cells was reduced, especially in the later period of culture. These AsA-overproducing cells also acquired resistance to paraquat (methyl viologen), which produces active oxygen species. These results contribute to the previous insights about AsA and raise the possibility of the generation of plants that have resistance to environmental stresses by increasing their AsA content. PMID- 15549374 TI - Molecular analysis of the CRINKLY4 gene family in Arabidopsis thaliana. AB - The maize (Zea mays L.) CRINKLY4 (CR4) gene encodes a serine/threonine receptor like kinase that controls an array of developmental processes in the plant and endosperm. The Arabidopsis thaliana (L.) Heynh. genome encodes an ortholog of CR4, ACR4, and four CRINKLY4-RELATED (CRR) proteins: AtCRR1, AtCRR2, AtCRR3 and AtCRK1. The available genome sequence of rice (Oryza sativa L.) encodes a CR4 ortholog, OsCR4, and four CRR proteins: OsCRR1, OsCRR2, OsCRR3 and OsCRR4, not necessarily orthologous to the Arabidopsis CRRs. A phylogenetic study showed that AtCRR1 and AtCRR2 form a clade closest to the CR4 group while all the other CRRs form a separate cluster. The five Arabidopsis genes are differentially expressed in various tissues. A construct formed by fusion of the ACR4 promoter and the GUS reporter, ACR4::GUS, is expressed primarily in developing tissues of the shoot. The ACR4 cytoplasmic domain functions in vitro as a serine/threonine kinase, while the AtCRR1 and AtCRR2 kinases are not active. The ability of ACR4 to phosphorylate AtCRR2 suggests that they might function in the same signal transduction pathway. T-DNA insertions were obtained in ACR4, AtCRR1, AtCRR2, AtCRR3 and AtCRK1. Mutations in acr4 show a phenotype restricted to the integuments and seed coat, suggesting that Arabidopsis might contain a redundant function that is lacking in maize. The lack of obvious mutant phenotypes in the crr mutants indicates they are not required for the hypothetical redundant function. PMID- 15549375 TI - Chrysanthemum stunt viroid disturbs the photoperiodic response for flowering of chrysanthemum plants. AB - Chrysanthemum ( Dendranthema grandiflorum Kitam.) is one of the qualitative short day flowering plants. Therefore, the flowering of chrysanthemum can usually be controlled by photoperiod. However, it was noted that 'Piato' plants infected by the chrysanthemum stunt viroid (CSVd) flowered autonomously even under long-day conditions. In this study, CSVd-free and CSVd-infected plants were prepared by culturing different-sized dissected shoot apical meristems (SAMs) of 'Piato'. Using these CSVd-free and CSVd-infected plants, we clarified the relationship between CSVd infection and the autonomous flowering of 'Piato'. Under natural short-day conditions, the flowering of plants regenerated from SAMs containing leaf primordia (LPs) was 1 month earlier than plants regenerated from LP-free SAMs. CSVd was detected from these early flowering plants by reverse transcription-polymerase chain reaction. On the other hand, CSVd was not detected in plants regenerated from LP-free SAMs. CSVd-infected and CSVd-free plants were grown under long-day conditions simulated by night-break lighting at 22:00 p.m. to 2:00 a.m. All CSVd-infected plants flowered autonomously even under long-day conditions; on the other hand, CSVd-free chrysanthemum plants maintained their vegetative growth. When the CSVd-free plants were inoculated with CSVd by grafting them to CSVd-infected rootstocks, they flowered autonomously even under night-break lighting. In this study, the results suggest that CSVd may control the qualitative development process, flowering, i.e. CSVd can induce the autonomous flowering of chrysanthemum. PMID- 15549376 TI - Expression and localization of cartilage-specific matrix protein chondromodulin-I mRNA in salivary pleomorphic adenomas. AB - Pleomorphic adenoma is the most common epithelial tumor in the salivary glands. This tumor frequently exhibits "mesenchyme"-like components, including myxoid or chondroid areas. Recently, using immunohistochemical techniques, we reported that cartilage-specific matrix protein, chondromodulin-I (ChM-I), was deposited on the inter-territorial matrix of the chondroid area in salivary pleomorphic adenomas and that ChM-I, which is also a strong angio-inhibitory factor, plays an important role in the avascular nature of the chondroid area and the chondroid formation in this type of tumor. To elucidate which cells express ChM-I mRNA in pleomorphic adenomas, we examined the expression and localization of ChM-I mRNA in this type of tumor using an in situ hybridization technique. Immunoreactivity for ChM-I was observed in the inter-territorial matrix of the chondroid area, especially around the lacunae, and in the cytoplasm of neoplastic myoepithelial cells of the myxoid element of pleomorphic adenomas. On in situ hybridization analysis, strong signals for ChM-I mRNA were detected in the cytoplasm of the lacuna cells of the chondroid element, and moderate to marked signals were observed in the cytoplasm of the neoplastic myoepithelial cells of the myxoid element. Signals for ChM-I mRNA were also seen in the cytoplasm of the spindle shaped neoplastic myoepithelial cells in the transitional areas between the myxoid and chondroid elements of this tumor. Signals for ChM-I mRNA were not seen in the inner ductal cells or the fibrous element. These findings indicate that lacuna cells and neoplastic myoepithelial cells express ChM-I mRNA and that mature ChM-I, which lacuna cells and neoplastic myoepithelial cells translate, is deposited in the chondroid matrix of pleomorphic adenomas. In conclusion, lacuna cells and neoplastic myoepithelial cells express ChM-I mRNA ectopically in pleomorphic adenoma, and this plays an important role in chondroid formation and hypovascularity in this type of tumor. PMID- 15549377 TI - The "wait and see" approach of acute otitis media. PMID- 15549378 TI - Treatment and outcome of severe and non-severe acute otitis media. AB - To determine outcomes in acute otitis media (AOM) according to severity of disease and to assess different initial treatment regimens, 308 with AOM were enrolled and divided into severe (n = 277; 89.9%) and non-severe (n = 31; 10.1%) groups based on symptoms and tympanic membrane changes. Children in the severe group were initially managed with amoxicillin (AMPC) whereas children in the non severe group were initially managed without antibiotics. Children were monitored on days 1, 5, 10, 14 and 28. Five outcome measures were assessed: disappearance of symptoms at day 5, resolution of tympanic membrane changes by day 28, disappearance of middle ear effusions by day 28, recurrence of acute symptoms prior to day 28, and need to change treatment regimens. Children with severe disease were more often male (57% versus 36%, P < 0.05) and more often colonized with pathogens (77% versus 55%, P < 0.05 than children with non-severe disease. The two groups were similar with respect to age and day care attendance. Despite differences in initial treatment regimens between the two groups, symptoms improved at the same rate for severe and non-severe disease, 94% by day 5. In contrast, tympanic membranes returned to normal in 69% of the severe and 81% of the non-severe groups by day 28; however, as early as day 5, 10% of the severe and 55% of the non-severe groups demonstrated normal tympanic membranes. Middle ear effusions similarly disappeared more slowly in the severe group, 52% versus 74% by day 14 and 76% versus 84% by day 28. Recurrence rates of acute symptoms occurred with equal frequency in the severe, 15%, and non-severe groups, 10%. Failure of the symptoms or the tympanic membranes to improve led to antibiotic changes in 59.9% of the severe group and to the addition of antibiotics in 51.6% of the non-severe group. Children in the severe group who failed to improve with an initial course of amoxicillin were younger (40.2 months versus 45.8 months, P < 0.05), had higher tympanic membrane scores (4.5 versus 4.1, P < 0.05), and were more often colonized with penicillin-resistant Streptococcus pneumoniae (33.8% versus 6.5%, P < 0.01) than children who responded to AMPC. In a similar manner, children with non-severe disease who failed to improve without antibiotics were younger (40.7 months versus 54.8 months, P < 0.05) and more often colonized with pathogens (75.0% versus 33.4%, P < 0.05). CONCLUSION: Severe disease occurred more often among males and among children colonized with pathogens. Response to treatment was impaired in younger children and in children colonized with pathogens, especially penicillin-resistant Streptococcus pneumoniae. PMID- 15549379 TI - Etanercept plus colchicine treatment in a child with tumour necrosis factor receptor-associated periodic syndrome abolishes auto-inflammatory episodes without normalising the subclinical acute phase response. AB - We investigated the cause of hereditary periodic fever syndrome in a Spanish child with recurrent long episodes of fever, migratory skin rash, myalgia, arthralgia, conjunctivitis and abdominal pain. Infectious and autoimmune causes were ruled out. No familial history was reported. Analysis of the tumour necrosis factor receptor superfamily 1A (TNFRSF1A) gene identified a missense mutation (G36E) on exon 3. The absence of this variant in the patient's parents and in controls identified it as a de novo disease-associated mutation. Clinical symptoms disappeared with administration of etanercept; however, levels of acute phase reactants remained increased and could not be stabilised by the addition of colchicine. We believe that this patient gained some symptomatic relief with etanercept therapy, although not enough to completely avoid the risk of amyloidosis. Thus it is debatable whether etanercept alone or combined with other drugs, is the treatment of choice for patients with tumour necrosis factor receptor-associated periodic syndrome. CONCLUSION: Since there is variability in treatment responses among different patients with tumour necrosis factor receptor associated periodic syndrome, we suggest that a systematic evaluation of acute phase reactants, especially SAA-1, could be useful in maintaining or modifying a given therapeutic approach in these patients. PMID- 15549380 TI - Colostrum from healthy Brazilian women inhibits adhesion and contains IgA antibodies reactive with Shiga toxin-producing Escherichia coli. AB - Although Shiga toxin-producing Escherichia coli (STEC) has been isolated in Brazil, severe manifestations of the infection, such as haemorrhagic colitis and haemolytic-uraemic syndrome, are extremely rare in our population. Enteropathogenic Escherichia coli (EPEC) is the main aetiological agent of acute infantile diarrhoea in Brazil. There are many similarities between STEC and EPEC, such as the ability to produce attaching and effacing (A/E) lesions and some virulence-associated factors. Our aim was to investigate the presence of anti STEC antibodies in healthy people living in an EPEC endemic area. Colostrum samples collected from 51 women living in low socio-economic conditions were analysed. Two STEC strains: O111:H- (Stx1) and O157:H7 (Stx2), and one EPEC strain (O111:H-) were used in the bacterial adhesion assays to HEp-2 cells, in the Stx1 and Stx2 cytotoxicity assays on Vero cells, in immunoblotting and in ELISA assays. All the samples strongly inhibited the adhesion of the three strains and contained SIgA antibodies reactive with antigens of EPEC O111:H-, STEC O111:H- and STEC O157:H7, mainly STEC and EPEC 94 kDa adhesin intimin. High titres of anti-LPS O111 antibodies were found in many samples. Nevertheless, the cytotoxic effect of both Stx1 and Stx2 on Vero cells was not neutralised by any sample. CONCLUSION: Our results suggest that Brazilian people may be exposed to Shiga toxin-producing Escherichia colimore frequently than previously thought or alternatively there may be a cross reactive immunity between enteropathogenic Escherichia coliand Shiga toxin-producing Escherichia coli. PMID- 15549381 TI - Helicobacter pylori and autoimmune hepatitis. PMID- 15549382 TI - Is ultrasound a useful adjunct for assessing malaria patients? AB - The value of ultrasonography as an adjunct for diagnosis and monitoring malaria was investigated. In all, 118 patients (male/female 65/53; age 2-78 years, median 29 years) with malaria underwent a standardised abdominal ultrasound examination at baseline. In 62 out of 118 patients, ultrasonography was repeated 21 days later. In the results at baseline, huge splenomegaly with firm organ consistency, consistent with hyperreactive malarious splenomegaly syndrome, was observed in two Cameroonese children. In the other 116 patients, the most common finding was non-specific splenomegaly (96/116, 82.76%), occurring more frequently in non immune patients (71/78, 91.03%) than in patients who had grown up in malaria endemic areas (25/38, 65.79%; P<0.002). No correlation was found between liver or spleen size and any clinical parameter. The results on day 21 show that, although splenomegaly after therapy persisted more frequently in patients with malaria recrudescence or relapse (8/8, 100%) than in patients cured (32/54, 59.26%; P<0.0421), the practical value of this finding is questionable. Ultrasonography cannot be regarded as a first-line diagnostic method in patients with malaria. PMID- 15549383 TI - The identification of a sequence related to apicomplexan enolase from Sarcocystis neurona. AB - Equine protozoal myeloencephalitis (EPM) is a neurological disease caused by Sarcocystis neurona, an apicomplexan parasite. S. neurona is also associated with EPM-like diseases in marine and small mammals. The mechanisms of transmission and ability to infect a wide host range remain obscure; therefore, characterization of essential proteins may provide evolutionary information allowing the development of novel chemotherapeutics that target non-mammalian biochemical pathways. In the current study, two-dimensional electrophoresis and matrix assisted laser desorption ionization-time of flight (MALDI-ToF) mass spectrometry were combined to characterize and identify an enolase protein from S. neurona based on peptide homology to the Toxoplasma gondii protein. Enolase is thought to be a vestigial, non-photosynthetic protein resulting from an evolutionary endosymbiosis event of an apicomplexan ancestor with green algae. Enolase has also been suggested to play a role in parasite stage conversion for T. gondii. Characterization of this protein in S. neurona and comparison to other protozoans indicate a biochemical similarity of S. neurona enolase to other tissue-cyst forming coccidians that cause encephalitis. PMID- 15549384 TI - Technical variability and required sample size of helminth egg isolation procedures: revisited. AB - Mes [Vet Parasitol (2003) 115:311-320] recently reported a quantitative study of repeated measurements of nematode egg counts in faecal samples from dairy cattle, in order to compare the faecal egg counts resulting from two different laboratory techniques, the widely used McMaster method and a newer salt-sugar flotation (SSF) method. He concluded that the SSF technique requires much smaller sample sizes, and is also potentially simpler to carry out, making it the method of choice. Here I re-analyse these data to show that if the comparison is done on the most appropriate measurement scale (lognormal), and the large difference in multiplication factors is taken into account, then there is little to choose between the McMaster and SSF techniques as far as the required sample size is concerned. In particular, the treatment of the data as normally rather than lognormally distributed leads to incorrect statistical tests, power analyses and confidence intervals. PMID- 15549385 TI - Natural infestation of the chewing lice (Werneckiella equi) on horses and treatment with imidacloprid and phoxim. AB - Infestation with the chewing louse (Werneckiella (Damalinia) equi) can be found on horses world-wide. Louse infestations, including clinical signs of louse derived dermatitis, are known from Icelandic horses. A clinical field investigation was conducted in Iceland using horses with natural louse infestations to evaluate the efficacy of imidacloprid in a 10% solution in comparison with phoxim in a 0.05% solution. A total of 27 horses received a single imidacloprid treatment using 16 ml of the 10% solution along the mane and on the dorso-lateral trunk. A further 43 horses were treated twice, 14 days apart, with phoxim, using 2 x 50 ml solution applied along the mane and the dorso lateral trunk. At the final evaluation on day 28, complete control of the lice was obtained for the imidacloprid treated horses and only a single moribund louse was found on two horses treated with phoxim. PMID- 15549386 TI - Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil. V. Cytokine concentrations in experimentally infected Wistar rats. AB - Tungiasis is caused by the penetration of the female sand flea Tunga penetrans into the skin of its host. This parasitic skin disease is almost invariably associated with an intense inflammation around embedded fleas, the underlying mechanisms being unknown. A study was undertaken to determine whether Wistar rats can be used as an animal model to assess cytokine kinetics during the natural course of the infection. Laboratory-raised Wistar rats were exposed in cages put on the soil in an area with high human attack rates. Rats were examined daily and blood samples were taken before exposure and at 2, 6, 10, 13, 16 and 20 days after flea penetration. TNF-alpha, IL-1 beta, IFN-gamma, IL-4, IL-10 and CINC (a rat cytokine- induced neutrophil chemoattractant and member of the IL-8 family) were determined by enzyme immunoassay. The results showed an increasing serum concentration of TNF-alpha and IL-1 beta 10-13 days after penetration and a rapid increase in IL-4 2 days after fleas became embedded. During the natural course of the infection, the ratio of the serum concentration of TNF-alpha to that of IL-10 decreased, indicating a relative increase in the secretion of the anti inflammatory cytokine. The treatment of lesions with silicone oil abrogated the natural disease course and changed the pattern of cytokine secretion. We conclude that the Wistar rat is an appropriate model to study immune responses in tungiasis. PMID- 15549387 TI - Comparative studies on intestine ultrastructure of third-stage larvae and adults of Cystidicoloides ephemeridarum (Nematoda, Cystidicolidae). AB - The intestinal epithelium of third-stage larvae and adults of Cystidicoloides ephemeridarum from haemocoel of mayflies and stomach of brown trout was studied by electron microscopy and cytochemistry. In section, the intestine of both stages is composed of a single layer of about ten undifferentiated intestinal cells in a ring. A labyrinth of deep invaginations is present in the basal region of each cell. The apical surface is modified into well developed, regularly arranged microvilli. These, together with numerous organelles engaged in metabolism and a well defined gut lumen filled with unidentifiable material suggest that the intestine may function in digestion and absorption during both stages. The adults seem to feed upon the semifluid content of the stomach of brown trout. Fortuitous oral infection with undetermined bacteria in vitro led to degenerative changes in the intestinal tissue and probably caused death of the infected specimens. Up to 75% of the cell volume in the L(3) is occupied by glycogen deposits. In the adults, a minor portion of glycogen, together with lipid droplets, has been observed. The adults are considered to rely more on aerobic metabolism, whereas anaerobic metabolism (glycolysis) may prevail in L(3). PMID- 15549388 TI - Co-reactivity of plasmodial histidine-rich protein 2 and aldolase on a combined immuno-chromographic-malaria dipstick (ICT) as a potential semi-quantitative marker of high Plasmodium falciparum parasitaemia. AB - The combined immuno-chromographic-malaria dipstick (ICT) for the rapid diagnosis of malaria detects both Plasmodium falciparum (P.f.)-specific, histidine-rich protein 2 (HRP-2) and a plasmodial aldolase expressed by all Plasmodium species pathogenic to humans. ICT was applied in 674 febrile returnees from malaria endemic regions attending our Tropical Diseases Unit. Microscopy confirmed malaria in 69/674 cases, of whom 67/69 had returned from Africa or Madagascar, and 2/69 from the Caribbean. Monoparasitic P.f. infection occurred in 52/69, mixed infection was due to P.f.+ P. ovale (P.o.) in 3/69, and P.f.+P. malariae (P.m.) in 1/69 cases. Monoparasitic P. vivax (P.v.) infection occurred in 8/69 , P.o. in 3/69, and P.m. in 2/69 cases . Whereas a positive HRP-2 band on the test was a highly sensitive indicator for P.f. infection (52/52 patients; sensitivity 100%), this was not the case for a positive aldolase band (25/52 patients; sensitivity 48.1%). Sensitivity of aldolase band for non-falciparum plasmodia was even lower: aldolase was positive in only 3/8 (37.5%) of patients with vivax malaria, and in 0/5 cases with P.o.- or P.m. infection. Co-reaction of both bands occurred more frequently in patients with P.f. parasitaemia of > or =40,000/microl (20/25, 80.0%) as compared to patients with P.f. parasitaemia <40,000/microl (5/27, 18.5%; P<0.00005), and to patients with mixed infection (P.f.+ P.o., P.f.+ P.m.: 2/4, 50.0%; diff. n.s.). In our series, co-reaction of HRP-2 and aldolase indicated monoparasitic falciparum malaria with high P.f. parasitaemia, rather than mixed infection. Whereas the aldolase band is not a reliable qualitative marker for malaria, co-reaction of HRP-2 and aldolase band may have a potential for indicating high parasitaemia in falciparum malaria. PMID- 15549389 TI - Posttranslational modifications of alpha-tubulin of Toxoplasma gondii. AB - The posttranslational modifications of alpha-tubulin of Toxoplasma gondii were characterized by antibodies and biochemical analysis of the carboxy-terminal peptide. Alpha-Tubulin is acetylated and glutamylated. Side chains with up to three glutamate residues are linked to Glu445 of T. gondii alpha-tubulin. The data suggest that the site of glutamylation on alpha-tubulin is conserved over a broad range of species. PMID- 15549390 TI - Borrelia burgdorferi infection prevalences in questing Ixodes ricinus ticks (Acari: Ixodidae) in urban and suburban Bonn, western Germany. AB - From March to October 2003, a total of 2,518 host-seeking Ixodes ricinus ticks (1,944 nymphs, 264 females, 310 males) were collected by blanket dragging at 45 sites all over the city area of Bonn, western Germany, to be checked for Borrelia burgdorferi infection. The collection sites included 20 private gardens, nine public recreational parks, the boundaries of 14 sylvatic suburban areas and two footpaths between suburban farmed fields. Generally, numbers of specimens collected along sylvatic suburban areas and at urban sites with dense tree populations were significantly higher than at the other collection sites. Out of 1,394 specimens (865 nymphs, 241 females, 288 males) that were randomly chosen for Borrelia analysis by a simple PCR, 250 (17.9 %) were found to be infected with B. burgdorferi sensu lato. While the infection prevalences varied significantly between females (26.6%), males (12.5%) and nymphs (17.3%), there were no striking differences between sylvatic and unwooded sites. A total of 92.8% of the ticks Borrelia-positive by the simple PCR were also positive in a diagnostic nested PCR. Using genospecies-specific oligonucleotide probes, single Borrelia genospecies infections (91.4%) could be assigned to B. afzelii (39.5%), B. garinii (27.9%), B. burgdorferi sensu stricto (15.6%) and B. valaisiana (8.6%) by DNA hybridization. Various combinations of double infections were observed in 4.3% of the infected ticks. Another 4.3% of the Borrelia infections were untypeable. The B. burgdorferi genospecies distribution in the city area was shown to be variable from site to site and, even more, it was distinct from rural collection sites near Bonn. This is ascribed to a different spectrum of reservoir hosts. Taking into account the infection prevalences of host-seeking ticks in the forested surroundings of Bonn, our study demonstrates that the risk of acquiring Lyme disease after a tick bite in urban/suburban areas is comparably as high as in woodlands outside of the city. PMID- 15549391 TI - Listeria monocytogenes infection-dependent transfer of exogenously added DNA to fibroblast COS-1 cells. AB - The addition of double-stranded circular or linear DNA encoding EGFP (the Enhanced Green Fluorescent Protein) to a Listeria -containing infection medium resulted in up to 8.6% COS-1 cells expressing the reporter protein. The transfer of naked DNA into host cells upon infection by Listeria was found to be dependent on the ability of the bacteria to synthesize internalins and listeriolysin. Since no binding of DNA to the bacterial cells was detected, DNA uptake seems to be the consequence of the simultaneous entry of infection medium, and thus of naked DNA, via the phagosomes induced by the bacterium to facilitate its own entry into the host cells. PMID- 15549392 TI - Hereditary prostate cancer in Finland: fine-mapping validates 3p26 as a major predisposition locus. AB - In a recent genome-wide linkage (GWL) analysis of Finnish families at high risk for prostate cancer, we found two novel putative susceptibility loci at 3p25-p26 and 11q14. Here, we report the fine-mapping of these two critical regions at high resolution with 39 microsatellite markers in 16 families, including multiplex families that were not used in the GWL scan. The maximum multipoint HLOD was 3.39 at 3p26 and 1.42 at 11q14. The highest LOD scores were seen around markers D3S1270 and D3S4559 (alpha=0.89), covering approximately two megabases. The two known genes in this region CHL1 (cell adhesion molecule with homology to L1CAM) and CNTN6 (contactin 6) were screened for exonic mutations in the families showing the strongest linkage, but no disease-segregating sequence variants were observed. The recombination map pointed to a region proximal to the area of best linkage, suggesting that more genes may need to be investigated as candidates. These results provide strong evidence for the existence of a prostate cancer susceptibility gene at 3p26 in Finnish prostate cancer families. This locus has not been strongly linked with hereditary prostate cancer in other populations. However, the mildly positive 3p LOD scores in a recent GWL analysis of patients from the United States suggest that the locus may also be important in other populations. PMID- 15549393 TI - Linkage disequilibrium between microsatellite markers in the Swedish Sami relative to a worldwide selection of populations. AB - The pattern of linkage disequilibrium (LD) is affected by a number of factors, including population demography. High LD is seen in populations with a relatively limited and constant size, presumably because of genetic drift. We have examined the extent of LD among over 300 genome-wide pattern microsatellite loci in 29 populations from around the world. The pattern of LD varied between populations, with a larger extent of LD in populations with limited size relative to larger populations. In addition, the LD between 88 less well-spaced microsatellite markers from 10 different genomic regions was examined in the Sami compared with the general Swedish population. For these markers, increased LD extending up to 5 Mb was detected in the Sami. The amount of LD also differed between the chromosomal regions. The amount of LD in the Sami makes this population suitable for the mapping of complex genetic traits. PMID- 15549394 TI - The impact of MECP2 mutations in the expression patterns of Rett syndrome patients. AB - Rett syndrome (RTT), the second most common cause of mental retardation in females, has been associated with mutations in MeCP2, the archetypical member of the methyl-CpG binding domain (MBD) family of proteins. MeCP2 additionally possesses a transcriptional repression domain (TRD). We have compared the gene expression profiles of RTT- and normal female-derived lymphoblastoid cells by using cDNA microarrays. Clustering analysis allowed the classification of RTT patients according to the localization of the MeCP2 mutation (MBD or TRD) and those with clinically diagnosed RTT but without detectable MeCP2 mutations. Numerous genes were observed to be overexpressed in RTT patients compared with control samples, including excellent candidate genes for neurodevelopmental disease. Chromatin immunoprecipitation analysis confirmed that binding of MeCP2 to corresponding promoter CpG islands was lost in RTT-derived cells harboring a mutation in the region of the MECP2 gene encoding the MBD. Bisulfite genomic sequencing demonstrated that the majority of MeCP2 binding occurred in DNA sequences with methylation-associated silencing. Most importantly, the finding that these genes are also methylated and bound by MeCP2 in neuron-related cells suggests a role in this neurodevelopmental disease. Our results provide new data of the underlying mechanisms of RTT and unveil novel targets of MeCP2-mediated gene repression. PMID- 15549395 TI - Mitochondrial GTPase mitofusin 2 mutation in Charcot-Marie-Tooth neuropathy type 2A. AB - Charcot-Marie-Tooth disease (CMT) has been classified into two types, CMT1 and CMT2, demyelinating and axonal forms, respectively. CMT2 has been further subdivided into eight groups by linkage studies. CMT2A is linked to chromosome 1p35-p36 and mutation in the kinesin family member 1B-beta (KIF1B) gene had been reported in one pedigree. However, no mutation in KIF1B was detected in other pedigrees with CMT2A and the mutations in the mitochondrial fusion protein mitofusin 2 (MFN2) gene were recently detected in those pedigrees. MFN2, a mitochondrial transmembrane GTPase, regulates the mitochondrial network architecture by fusion of mitochondria. We studied MFN2 in 81 Japanese patients with axonal or unclassified CMT and detected seven mutations in seven unrelated patients. Six of them were novel and one of them was a de novo mutation. Most mutations locate within or immediately upstream of the GTPase domain or within two coiled-coil domains, which are critical for the functioning or mitochondrial targeting of MFN2. Formation of a mitochondrial network would be required to maintain the functional peripheral nerve axon. PMID- 15549396 TI - A novel 5q11.2 deletion detected by microarray comparative genomic hybridisation in a child referred as a case of suspected 22q11 deletion syndrome. AB - The 22q11 deletion syndrome (22q11DS) is a developmental syndrome comprising of heart, palate, thymus and parathyroid glands defects. Individuals with 22q11DS usually carry a 1.5- to 3-Mb heterozygous deletion on chromosome 22q11.2. However, there are many patients with features of 22q11DS without a known cause from conventional karyotype and FISH analysis. Six patients with features of 22q11DS, a normal chromosomal and FISH 22q11 analysis, were selected for investigation by microarray genomic comparative hybridisation (array CGH). Array CGH is a powerful technology enabling detection of submicroscopic chromosome duplications and deletions by comparing a differentially labelled test sample to a control. The samples are co-hybridised to a microarray containing genomic clones and the resulting ratio of fluorescence intensities on each array element is proportional to the DNA copy number difference. No chromosomal changes were detected by hybridisation to a high resolution array representing chromosome 22q. However, one patient was found to have a 6-Mb deletion on 5q11.2 detected by a whole genome 1-Mb array. This deletion was confirmed with fluorescence in-situ hybridisation (FISH) and microsatellite marker analysis. It is the first deletion described in this region. The patient had tetralogy of Fallot, a bifid uvula and velopharyngeal insufficiency, short stature, learning and behavioural difficulties. This case shows the increased sensitivity of array CGH over detailed karyotype analysis for detection of chromosomal changes. It is anticipated that array CGH will improve the clinician's capacity to diagnose congenital syndromes with an unknown aetiology. PMID- 15549397 TI - Developmental changes in the expression of nicotinic acetylcholine receptor alpha subunits in the rat heart. AB - Neuronal nicotinic acetylcholine receptors (nAChR) are ligand-gated ion channels that consist of various subunits. During ontogeny, muscular and neuronal nAChR undergo changes in the distribution and subunit composition in skeletal muscle and brain, respectively. Here, we have investigated the occurrence of the ligand binding alpha-subunits of neuronal nAChR by means of reverse transcription/polymerase chain reaction and immunohistochemistry in the rat heart during prenatal and postnatal development and after capsaicin-induced sensory denervation. mRNAs coding for the alpha4, alpha5, alpha7 and alpha10 subunits were detected throughout all developmental stages. Messenger coding for the alpha2 subunit was first detectable at developmental stage E20; alpha3 subunit mRNA was expressed throughout all prenatal developmental stages, whereas it was restricted postnatally to the atria. mRNA for alpha6 was observed at E14-P8 but was absent thereafter. At no developmental stage could an unequivocal signal for alpha9 nAChR subunit mRNA be obtained. The expression pattern was unchanged by capsaicin treatment. Immunohistochemistry demonstrated alpha7 subunits on cardiac neurons, fibroblasts and cardiomyocytes and alpha2/4 subunits on cardiomyocytes with a postnatal redistribution to intercalated discs, as shown by cryo immunoelectron microscopy. Our results indicate an additional non-neuronal expression of nAChR subunits in the rat heart that, as in skeletal muscle, precedes functional innervation and then undergoes changes in its distribution on the surface of cells. PMID- 15549398 TI - Cholesterol depletion by methyl-beta-cyclodextrin enhances myoblast fusion and induces the formation of myotubes with disorganized nuclei. AB - The formation of a skeletal muscle fiber begins with the withdrawal of committed mononucleated precursors from the cell cycle. These myoblasts elongate while aligning with each other, guided by recognition between their membranes. This step is followed by cell fusion and the formation of long striated multinucleated myotubes. We used methyl-beta-cyclodextrin (MCD) in primary cultured chick skeletal muscle cells to deplete membrane cholesterol and investigate its role during myogenesis. MCD promoted a significant increase in the expression of troponin T, enhanced myoblast fusion, and induced the formation of large multinucleated myotubes with nuclei being clustered centrally and not aligned at the cell periphery. MCD myotubes were striated, as indicated by sarcomeric alpha actinin staining, and microtubule and desmin filament distribution was not altered. Pre-fusion MCD-treated myoblasts formed large aggregates, with cadherin and beta-catenin being accumulated in cell adhesion contacts. We also found that the membrane microdomain marker GM1 was not present as clusters in the membrane of MCD-treated myoblasts. Our data demonstrate that cholesterol is involved in the early steps of skeletal muscle differentiation. PMID- 15549399 TI - Habitat quality as a predictor of spatial variation in blue tit reproductive performance: a multi-plot analysis in a heterogeneous landscape. AB - Vertebrate studies have rarely investigated the influence of spatial variation in habitat richness on both short-term (breeding) and long-term (offspring recruitment) reproductive performance using simultaneously multi-patch, multi habitat type and multi-year approaches at landscape level. Here we present results of such an approach using the influence of two oak tree (Quercus ilex, Q. humilis) species on reproductive performance in Corsican blue tits (Parus caeruleus ogliastrae) as a model system. We found that blue tits breeding in rich broad-leaved deciduous patches consistently laid eggs earlier in the season, and produced larger clutches and more fledglings of higher quality, than those breeding in poor evergreen patches. Also, parents, especially males, were in better physical condition in the broad-leaved deciduous than in the evergreen patches. Surprisingly, estimates of long-term effects of reproduction, such as recruitment rates of locally born offspring, did not differ between the two habitat types. Our results suggest that short-term breeding performance and phenotypic quality of both chicks and parents do not necessarily provide reliable information about contributions to following generations at a scale larger than that of the local study plot. Differences in reproductive performance between the two oak habitat types could not be attributed to density-dependent effects, differences in levels of nest predation, or differences in age structure of the birds. We suggest that habitats that are optimal for breeding are not necessarily optimal for survival after the breeding season. PMID- 15549400 TI - Effects of low-efficiency pollinators on plant fitness and floral trait evolution in Campanula americana (Campanulaceae). AB - Floral visitors vary in their pollination efficiency and their preferences for floral traits. If low-efficiency pollinators decrease the amount of pollen available to higher efficiency visitors, then low-efficiency visitors may actually have negative fitness consequences for the plants that they visit. We used experimental arrays in two populations to determine the floral preferences and the fitness effects of low-efficiency (or "ugly") pollinators on Campanula americana. These ugly pollinators (halictid bees) preferentially visited flowers with pollen over flowers that had had their pollen removed. C. americana pollen color varies quantitatively from light tan to dark purple, and we found that natural variation in pollen color influenced the magnitude of halictid preferences for flowers with pollen. In general, preferences for flowers with pollen were stronger when the ugly pollinators foraged in arrays of flowers with tan-colored pollen than in arrays with purple-colored pollen. When plants received few visits by efficient Bombus pollinators, visits by ugly pollinators significantly decreased siring success relative to plants where visits by ugly pollinators were prevented. In contrast, ugly pollinators did not influence siring success when higher efficiency pollinators were more abundant. Thus, the relationship between low-efficiency pollinators and the plants that they visit varies from commensalistic to antagonistic depending on the presence of other pollinators in the community. Our findings suggest that the negative fitness effects and floral preferences of low-efficiency or "ugly" pollinators may contribute to the maintenance of a pollen color polymorphism in C. americana. PMID- 15549401 TI - Rapid nutrient cycling in leaf litter from invasive plants in Hawai'i. AB - Physiological traits that contribute to the establishment and spread of invasive plant species could also have impacts on ecosystem processes. The traits prevalent in many invasive plants, such as high specific leaf areas, rapid growth rates, and elevated leaf nutrient concentrations, improve litter quality and should increase rates of decomposition and nutrient cycling. To test for these ecosystem impacts, we measured initial leaf litter properties, decomposition rates, and nutrient dynamics in 11 understory plants from the Hawaiian islands in control and nitrogen + phosphorus fertilized plots. These included five common native species, four of which were ferns, and six aggressive invasive species, including five angiosperms and one fern. We found a 50-fold variation in leaf litter decay rates, with natives decaying at rates of 0.2-2.3 year(-1) and invaders at 1.4-9.3 year(-1). This difference was driven by very low decomposition rates in native fern litter. Fertilization significantly increased the decay rates of leaf litter from two native and two invasive species. Most invasive litter types lost nitrogen and phosphorus more rapidly and in larger quantities than comparable native litter types. All litter types except three native ferns lost nitrogen after 100 days of decomposition, and all litter types except the most recalcitrant native ferns lost >50% of initial phosphorus by the end of the experiment (204-735 days). If invasive understory plants displace native species, nutrient cycling rates could increase dramatically due to rapid decomposition and nutrient release from invasive litter. Such changes are likely to cause a positive feedback to invasion in Hawai'i because many invasive plants thrive on nutrient-rich soils. PMID- 15549402 TI - Preferential uptake of soil nitrogen forms by grassland plant species. AB - In this study, we assessed whether a range of temperate grassland species showed preferential uptake for different chemical forms of N, including inorganic N and a range of amino acids that commonly occur in temperate grassland soil. Preferential uptake of dual-labelled (13C and 15N) glycine, serine, arginine and phenylalanine, as compared to inorganic N, was tested using plants growing in pots with natural field soil. We selected five grass species representing a gradient from fertilised, productive pastures to extensive, low productivity pastures (Lolium perenne, Holcus lanatus, Anthoxanthum odoratum, Deschampsia flexuosa, and Nardus stricta). Our data show that all grass species were able to take up directly a diversity of soil amino acids of varying complexity. Moreover, we present evidence of marked inter-species differences in preferential use of chemical forms of N of varying complexity. L. perenne was relatively more effective at using inorganic N and glycine compared to the most complex amino acid phenylalanine, whereas N. stricta showed a significant preference for serine over inorganic N. Total plant N acquisition, measured as root and shoot concentration of labelled compounds, also revealed pronounced inter-species differences which were related to plant growth rate: plants with higher biomass production were found to take up more inorganic N. Our findings indicate that species-specific differences in direct uptake of different N forms combined with total N acquisition could explain changes in competitive dominance of grass species in grasslands of differing fertility. PMID- 15549403 TI - Avian reproductive failure in response to an extreme climatic event. AB - Recently, climate change research has emphasized the potential increase in the frequency and severity of climatic extremes. We compared the reproductive effort and output among four species of passerine birds in coastal southern California, USA, a semi-arid region, during a normal precipitation year (2001) and the driest year in a 150-year climate record (2002). Both reproductive effort and output differed dramatically between years. Mean reproductive output among the four species was 2.37 fledglings/pair in 2001 and 88.4% of all pairs observed attempted at least one nest. The birds attempted a mean of 1.44 nests per pair and were successful in 47.7% of those attempts. In 2002, only 6.7% of the pairs even attempted a nest and only 1.8% were successful, for a total output of 0.07 fledglings per pair. The abundance of suitable arthropod prey items in the environment was also much lower in 2002, suggesting that low food availability was the proximal cause of the reproductive failure. The data for one of these species, the rufous-crowned sparrow (Aimophila ruficeps), were combined with reproductive and rainfall data from a previous 3-year study (1997-1999) in the same sites. The combined data sets suggest that the response of reproduction to rainfall variation is linear, and that the low end of the precipitation range brings the population near reproductive failure. Any change in climate that would increase the frequency of extreme dry conditions would likely endanger populations of these species. PMID- 15549404 TI - Tight coupling between leaf area index and foliage N content in arctic plant communities. AB - The large spatial heterogeneity of arctic landscapes complicates efforts to quantify key processes of these ecosystems, for example productivity, at the landscape level. Robust relationships that help to simplify and explain observed patterns, are thus powerful tools for understanding and predicting vegetation distribution and dynamics. Here we present the same linear relationship between Leaf area index (LAI) and Total foliar nitrogen (TFN), the two factors determining the photosynthetic capacity of vegetation, across a wide range of tundra vegetation types in both northern Sweden and Alaska between leaf area indices of 0 and 1 m2 m(-2), which is essentially the entire range of leaf area index values for the Arctic as a whole. Surprisingly, this simple relationship arises as an emergent property at the plant community level, whereas at the species level a large variability in leaf traits exists. As the relationship between LAI and TFN exists among such varied ecosystems, the arctic environment must impose tight constraints on vegetation canopy development. This relationship simplifies the quantification of vegetation productivity of arctic vegetation types as the two most important drivers of productivity can be estimated reliably from remotely sensed NDVI images. PMID- 15549405 TI - Leaf traits and associated ecosystem characteristics across subtropical and timberline forests in the Gongga Mountains, Eastern Tibetan Plateau. AB - Knowledge of how leaf characteristics might be used to deduce information on ecosystem functioning and how this scaling task could be done is limited. In this study, we present field data for leaf lifespan, specific leaf area (SLA) and mass and area-based leaf nitrogen concentrations (N(mass), N(area)) of dominant tree species and the associated stand foliage N-pool, leaf area index (LAI), root biomass, aboveground biomass, net primary productivity (NPP) and soil available-N content in six undisturbed forest plots along subtropical to timberline gradients on the eastern slope of the Gongga Mountains. We developed a methodology to calculate the whole-canopy mean leaf traits to include all tree species (groups) in each of the six plots through a series of weighted averages scaled up from leaf-level measurements. These defined whole-canopy mean leaf traits were equivalent to the traits of a leaf in regard to their interrelationships and altitudinal trends, but were more useful for large-scale pattern analysis of ecosystem structure and function. The whole-canopy mean leaf lifespan and leaf N(mass) mainly showed significant relationships with stand foliage N-pool, NPP, LAI and root biomass. In general, as elevation increased, the whole-canopy mean leaf lifespan and leaf N(area) and stand LAI and foliage N-pool increased to their maximum, whereas the whole-canopy mean SLA and leaf N(mass) and stand NPP and root biomass decreased from their maximum. The whole-canopy mean leaf lifespan and stand foliage N-pool both converged towards threshold-like logistic relationships with annual mean temperature and soil available-N variables. Our results are further supported by additional literature data in the Americas and eastern China. PMID- 15549406 TI - Hybrid process models for process optimisation, monitoring and control. AB - Hybrid models aim to describe different components of a process in different ways. This makes sense when the corresponding knowledge to be represented is different as well. In this way, the most efficient representations can be chosen and, thus, the model performance can be increased significantly. From the various possible variants of hybrid model, three are selected which were applied for important biotechnical processes, two of them from existing production processes. The examples show that hybrid models are powerful tools for process optimisation, monitoring and control. PMID- 15549407 TI - Proximal renal tubular dysfunction in primary distal renal tubular acidosis. AB - Low-molecular-weight (LMW) proteinuria has been described in patients with primary distal renal tubular acidosis (dRTA). However, other proximal renal tubular dysfunctions have rarely been reported. In this report we describe reversible and multiple proximal renal tubular cell dysfunctions in a patient with dRTA. A 4-year-old girl was admitted to our hospital for investigation of short stature and proteinuria. Laboratory studies revealed a hyperchloremic metabolic acidosis without aciduria, hypokalemia, hypouricemia with uricosuria, hypercalciuria, LMW proteinuria, phosphaturia, and generalized aminoaciduria. The patient was diagnosed as having dRTA with multiple proximal renal tubular dysfunctions. All proximal renal tubular dysfunction subsided 1.5 years after starting alkali therapy. The precise pathogenic mechanisms underlying the development of multiple proximal renal tubular dysfunctions in dRTA remained unclear. However, proximal renal tubular endosomal dysfunction resulting from a profound intracellular acidosis caused by vacuolar H+-ATPase dysfunction or hypokalemic nephropathy might contribute to the development of proximal renal tubular dysfunctions in patients with dRTA. PMID- 15549408 TI - A realistic chance for gene therapy in the near future. AB - The expanding knowledge of the genetic and cellular mechanisms of human diseases in the post-genomic era coupled with the development of different vector systems to efficiently transfer genes to a variety of cell types and organs in vivo gave rise to the concept of gene therapy as a promising therapeutic option for genetic and acquired diseases. Gene therapy has been the focus of both enthusiasm and critique in the past years. Major progress has been achieved in evaluating gene therapy in clinical trials. However, a number of hurdles must still be overcome to make gene therapy safe and applicable for human diseases. Increased knowledge of the interaction of the gene therapy vehicles with the host has resulted in modifications of existing and the development of new vector systems, as well as adjustments of future clinical applications. Adeno-associated virus vectors, retrovirus- and lentivirus-based vectors show great promise for the correction of monogenic diseases. Correction of the genetic defect can be attempted by either in vivo administration to directly target a diseased organ or by administration of ex vivo genetically modified cells, e.g., bone marrow stem cells. The lack of persistent expression and the immune responses of the host have limited the use of adenovirus vectors for the permanent correction of monogenic diseases. However, the ease of production and the number of cell types and organs that can be efficiently infected make adenovirus-based vectors a promising tool for applications where permanent gene expression is not the therapeutic goal or where the induction of immune responses is the desired response, as for genetic vaccines. Overall, gene therapy remains promising for the correction of genetic as well as acquired disorders, where permanent or transient expression of a gene product will be therapeutic. PMID- 15549409 TI - Necrotizing fasciitis in a child: a rare complication of idiopathic nephrotic syndrome. AB - In nephrotic syndrome there is an increased tendency for bacterial infections due to immunological changes secondary to proteinuria, treatment (including steroids), and other as yet unknown causes. However, necrotizing fasciitis (NF) is an uncommon complication of the disease and has rarely been reported in nephrotic children. We report a 14-month-old boy with nephrotic syndrome who developed sepsis and NF as a complication. He was treated successfully with intensive medical and surgical treatment. PMID- 15549410 TI - The multi-functional role of insulin-like growth factor binding proteins in bone. AB - The insulin-like growth factor (IGF) system is an important regulator of bone formation. The IGFs (IGF-I and IGF-II) are the most abundant growth factors produced by bone, and are regulated by their six high affinity binding proteins (IGFBPs). The IGFBPs are produced by osteoblasts and are responsible for transporting the IGFs and extending their half-lives. In general, IGFBP-1, -2, 4, and -6 inhibit and IGFBP-3 and -5 stimulate osteoblast function. IGFBP-4 and 5 are the most abundant IGFBPs produced by osteoblasts, and therefore they are the primary focus of this review. IGFBP-5 is an important stimulator of bone formation and may also function independently of IGFs. IGFBP-4 inhibits osteoblast function by sequestering IGF and preventing it from binding to its receptor. This review focuses on the specific IGF-dependent and IGF-independent roles of the IGFBPs in bone formation, as well as their potential mechanisms of action. In addition, discussion of the regulation of the IGFBPs by post translational modification (i.e., proteolysis) has been included. Studies on the regulation of production and actions of IGFBPs suggest that the IGFBP system in bone is pleiotropic and capable of serving multiple effector inputs from systemic and local sources. PMID- 15549411 TI - Alterations of the growth plate in chronic renal failure. AB - Chronic renal failure modifies the morphology and dynamics of the growth plate (GP) of long bones. In young uremic rats, the height of cartilage columns of GP may vary markedly. The reasons for this variation are unknown, although the severity and duration of renal failure and the type of renal osteodystrophy have been shown to influence the height of GP cartilage. Expansion of GP cartilage is associated with that of the hypertrophic stratum. The interference of uremia with the process of chondrocyte differentiation is suggested by some morphological features. However, analysis by immunohistochemistry and/or in situ hybridization of markers of chondrocyte maturation in the GP of uremic rats has yielded conflicting results. Thus, there have been reported normal and reduced mRNA levels for collagen X, parathyroid hormone/parathyroid hormone-related peptide receptor, and matrix metalloproteinase 9, as well as normal mRNA and protein expression for vascular endothelial growth factor and chondromodulin I, peptides related to the control of angiogenesis. In addition, a decreased immunohistochemical signal for growth hormone receptor and low insulin-like growth factor I mRNA in the proliferative zone of uremic GP are supportive of reduced chondrocyte proliferation. Growth hormone treatment improves chondrocyte maturation and activates bone metabolism in the primary spongiosa. PMID- 15549412 TI - Pathogenesis of parathyroid dysfunction in end-stage kidney disease. AB - Small decreases in serum calcium (Ca(2+)) and more-prolonged increases in serum phosphate (Pi) stimulate the parathyroid (PT) to secrete parathyroid hormone (PTH). 1,25-Dihydroxyvitamin D(3) [1,25(OH)(2) D(3)] decreases PTH synthesis and secretion. The prolonged decrease in serum Ca(2+) and 1,25(OH)(2) D(3), or increase in serum Pi, observed in patients with chronic renal failure leads to a secondary increase in serum PTH. This secondary hyperparathyroidism involves increases in PTH gene expression, synthesis, and secretion and, if chronic, to proliferation of the PT cells. A low serum Ca(2+) leads to an increase in PTH secretion, PTH mRNA stability, and PT cell proliferation. Pi also regulates the PT in a similar manner. The effect of Ca(2+) on the PT is mediated by a membrane Ca(2+) receptor. 1,25(OH)(2) D(3) decreases PTH gene transcription. Ca(2+) and Pi regulate the PTH gene post transcriptionally by regulating the binding of PT cytosolic proteins, trans factors, to a defined cis sequence in the PTH mRNA 3' untranslated region, thereby determining the stability of the transcript. The PT trans factors and cis elements have been defined. PMID- 15549413 TI - Cardiovascular disease as a late complication of end-stage renal disease in children. AB - As in older adults, cardiovascular disease is the most important cause of death in adolescents and young adult patients with end-stage renal disease (ESRD) since childhood. This concerns patients on dialysis as well as transplant patients, despite the fact that a long duration of dialysis during childhood is an extra mortality risk factor. Left ventricular hypertrophy (LVH), aortic valve calcification, and increased arterial stiffness, but not increased arterial intima media thickening, are the most frequently observed alterations in young adult survivors with childhood ESRD. In transplanted patients a concentric LVH as a result of chronic hypertension is mostly observed; in dialysis patients a more asymmetric septal LVH is found as a result of chronic volume overload. These results suggest that in children and young adults with ESRD chronic pressure and volume overload, a high calcium-phosphate product, and chronic inflammation, but not dyslipidemia, play a role in the development of cardiovascular disease. PMID- 15549414 TI - Impact of inflammation and oxidative stress on vascular calcifications in chronic kidney disease. AB - Vascular and/or valvular calcifications in patients with chronic kidney disease (CKD) appear to indicate a poor prognosis in terms of overall survival and cardiovascular morbidity and mortality. Inflammation and oxidative stress represent new features of the arterial and/or valvular calcification process. However, only limited observational and epidemiological data are available in these areas. Therefore, the link between inflammation, oxidation and vascular and/or valvular calcifications deserves careful consideration in CKD patients, since they may become targets for the development of new therapeutic strategies. PMID- 15549415 TI - "Missing" inhibitors of calcification: general aspects and implications in renal failure. AB - In the recent past, it has become increasingly clear that extracellular calcium and phosphate homeostasis is a tightly regulated process. Since the physiological serum concentrations of calcium and phosphate are several orders of magnitude above their solubility product, mechanisms inhibiting precipitation must be operative to prevent extraosseous calcification. A number of local and systemic calcification inhibitors, including fetuin-A, matrix Gla protein, and osteoprotegerin, have been identified in recent years. Deficiency and dysregulation of such factors may contribute to morbidity and even mortality. Extraosseous calcifications occur with high prevalence in patients with end-stage renal disease. In particular, vascular manifestations are clearly associated with cardiovascular events and decreased survival. In addition to the well-established roles of hyperphosphatemia and an increased calcium x phosphate product, the biological and potential clinical roles of disturbances in calcification inhibition in uremia are discussed in this overview. PMID- 15549416 TI - Allelic differences in a quantitative trait locus affecting insulin-like growth factor-I impact skeletal acquisition and body composition. AB - Insulin-like growth factor-I (IGF-I) is critical for optimal skeletal growth and maintenance. Knockout and transgenic models have provided significant insights into the role of IGF-I in bone modeling and remodeling. Congenic mice demonstrate allelic differences in particular quantitative trait loci (QTL). One such model is congenic 6T, which contains a QTL for reduced serum IGF-I donated from C3H/HeJ on a pure C57Bl/6 J (B6) background. In this study we found a 30%-50% reduction in IGF-I expression in bone, liver, and fat of the congenic 6T mouse, as well as lower circulating IGF-I compared with control B6. 6T mice also had a greater percentage body fat, but reduced serum leptin. These changes were associated with reduced cortical and trabecular bone mineral density, impaired bone formation but no change in bone resorption. Moreover, the anabolic skeletal response to intermittent parathyroid hormone (PTH) therapy was blunted in 6T compared with B6, potentially in response to greater programmed cell death in osteocytes and osteoblasts of 6T. In summary, allelic differences in IGF-I expression impact peak bone acquisition and body composition, as well as the skeletal response to PTH. Lifelong changes in circulating and skeletal IGF-I may be relevant for the pathophysiology of several diseases, including chronic renal failure. PMID- 15549417 TI - Cytokine inhibition of JAK-STAT signaling: a new mechanism of growth hormone resistance. AB - Growth hormone (GH) and insulin-like growth factor (IGF)-I are potent regulators of muscle mass in health and disease. This somatomedin axis is markedly deranged in various catabolic conditions in which circulating and tissue levels of inflammatory cytokines are elevated. The plasma concentration of IGF-I, which is primarily determined by hepatic synthesis and secretion of the peptide hormone, is dramatically decreased during catabolic and inflammatory conditions. Moreover, many of these conditions are also associated with an inability of GH to stimulate hepatic IGF-I synthesis. This defect results from an impaired phosphorylation and activation of the traditional JAK2/STAT5 signal transduction pathway. Numerous lines of evidence support the role of tumor necrosis factor (TNF)-alpha as a prominent but probably not the sole mediator of the sepsis-induced impairment in basal and GH-stimulated IGF-I synthesis in liver. Additionally, catabolic conditions produce comparable alterations in skeletal muscle. However, in contrast to liver, the GH resistance in muscle is not mediated by a defect in STAT5 phosphorylation. Muscle is now recognized to respond to infectious stimuli with the production of numerous inflammatory cytokines, including TNF-alpha. Furthermore, myocytes cultured with TNF-alpha are GH resistant and this defect appears mediated via a STAT5-independent but JNK-dependent mechanism. Collectively, these changes act to limit IGF-I availability in muscle, which disturbs protein balance and results in the loss of protein stores in catabolic and inflammatory conditions. PMID- 15549419 TI - [Chronic radiculopathy. Use of minimally invasive percutaneous epidural neurolysis according to Racz]. AB - Treatment of chronic low back pain exhibiting radicular symptoms poses a clinical problem that has not yet been solved. The technique of percutaneous minimally invasive neurolysis described by Racz is being performed increasingly to treat chronic radiculopathy. A total of 61 patients with corresponding symptomatology after screening for inclusion and exclusion criteria in the region of the lumbar spinal nerve were treated with the Racz catheter technique. Distinct clinical improvement was observed at the 3- and 6-month follow-ups after percutaneous minimally invasive epidural neurolysis. Subjective pain perception, quantified by the McNab score, clearly improved after 3 as well as 6 months. With the exception of partial catheter shearing in two cases and one occurrence of infection, no relevant side effects were noted. The Racz catheter technique for treatment of chronic radiculopathy following disk surgery is suitable with minimal side effects. PMID- 15549418 TI - Molecular physiology, pathology, and regulation of the growth hormone/insulin like growth factor-I system. AB - Since the somatomedin hypothesis of growth hormone (GH) action was first formulated nearly 50 years ago, the key roles of both GH and insulin-like growth factor (IGF)-I in human growth have been confirmed and extended to include local effects on tissue maintenance and repair. More recent insights have revealed a dark side to the GH/IGF-I signaling system. Both proteins have been implicated as potential contributing factors in selected human cancers, and normal activity through this signaling pathway has been linked to diminished lifespan in experimental animals. This review highlights both the positive and negative aspects of the GH/IGF-I-growth pathway. The overall goal is to reinforce the need for more complete understanding of the mechanisms of signaling and action of GH and IGF-I, in order to separate, if possible, the potentially beneficial outcomes on growth and on tissue maintenance and repair from deleterious effects on cancer risk and lifespan. PMID- 15549420 TI - [Cannabinoids--signal transduction and mode of action]. AB - The therapeutic use of cannabinoids, the components of cannabis sativa L., was investigated in numerous researches in detail. Animal studies revealed that cannabinoid receptor agonists alter pain-associated behaviour, have immune suppressive properties, suppress tumor growth, modulate sensitisation processes and influence memory and learning. Those effects are mediated by two membrane bound cannabinoid receptors and as mechanisms of signal transduction blockade of ion channels, inhibition of adenylate cyclase and retrograde inhibition of neurotransmitter release are currently being discussed. In clinical studies oral administration of cannabinoids indicated beneficial results during the therapy of multiple sclerosis, weight loss, nausea and vomiting due to chemotherapy, and intractable pruritus. However, therapy of chronic pain conditions revealed conflicting results and unequivocal success could not have been delivered due to unwanted side effects. Further multicentre studies are required to estimate cannabinoids as novel therapeutic tools for the treatment of chronic pain. PMID- 15549422 TI - Mortality and displaced mortality during heat waves in the Czech Republic. AB - The aims of this study were to assess impacts of hot summer periods on mortality in the Czech Republic and to quantify the size of the short-term displacement effect which resulted in lower than expected mortality after heat waves. The analysis covered the period 1982-2000 when several extraordinarily hot summers occurred in central Europe. Daily total all-cause mortality and mortality due to cardiovascular diseases (CVD) in the entire population of the Czech Republic (approximately 10 million inhabitants) were examined. The daily death counts were standardized to account for the long-term decline in mortality and the seasonal and weekly cycles. Heat-related mortality is better expressed if 1-day lag after temperature is considered compared to the unlagged relationship. With the 1-day lag, both excess total mortality and excess CVD mortality were positive during all 17 heat waves, and in 14 (12) heat waves the increase in total (CVD) mortality was statistically significant ( P=0.05). The mean relative rise in total mortality during heat waves was 13%. The response was greater in females than males and similar regardless of whether total or CVD mortality was used. The largest relative increases, exceeding 20% in both total and CVD mortality, were associated with heat waves which occurred in early summer (the first half of July 1984 and June 1994). The mortality displacement effect played an important role since mortality tended to be lower than expected after hot periods. The mean net mortality change due to heat waves was estimated to be about a 1% increase in the number of deaths. The large relative increases during some heat waves were particularly noteworthy since the study (in contrast to most analyses of the heat stress/mortality relationship) was not restricted to an urban area and/or an elderly population. PMID- 15549421 TI - Role of afferent pathways of heat and cold in body temperature regulation. AB - The detection of surface and internal temperatures is achieved by axons terminating at lamina I of the spinal dorsal horn, otherwise approached only by nociceptive afferents. Recent advances in thermal physiology research have disclosed that temperature-sensitive ion channels belonging to the "transient receptor potential" family exist in the peripheral sensory neurons and in the brain. Thermosensory, nociceptive and polymodal afferents project to different thalamic nuclei, and specific pathways to the insular cortex evoke the conscious experience of thermal sensation. The posterior insular region represents discriminative thermal sensation, while the largest correlation with subjective ratings of temperature is located in the orbitofrontal and anterior insular cortex. The insular cortex forms an integrative part of the limbic system and is closely tied with the hypothalamus, the amygdala, the anterior cingulate cortex and the orbitofrontal cortex and emerges as the main coordinator of behavioral, autonomic and endocrine responses to both non-noxious and noxious thermal stimuli. The firing rate of warm and cold receptors is not altered by pyrogens. A strong correlation between the onset of fever and production of superoxide by macrophages following the injection of pyrogens implicates reactive oxygen species as elicitors of fever, a hypothesis strengthened by the observation that oxygen radical scavengers or thiol reductants act as antipyretics. Oxidative stress appears to be sensed by the brain and a likely structure for its detection may be the redox-sensitive site of the N-methyl-D-aspartate (NMDA) receptor for glutamate, in that oxidation of this site causes fever while its reduction lowers body temperature, effects which are abrogated by specific NMDA receptor blockers. PMID- 15549424 TI - Comparison of proposed diagnostic criteria with FACT-F and VAS for cancer-related fatigue: proposal for use as a screening tool. AB - The objective was to validate the use of the proposed International Statistical Classification of Diseases and Related Health Problems (10th revision) (ICD-10) criteria for fatigue (P-ICD10) through comparison with the Functional Assessment of Cancer Therapy Fatigue (FACT-F) subscale and three visual analogue scale (VAS) qualities in cancer patients thought to be fatigued. Fatigue was assessed in 834 cancer patients at three clinical centres in Belgium, using P-ICD10, FACT-F, and VAS to assess: level of energy (VAS1), quality of life (VAS2), and ability to perform daily activities (VAS3). Of the 834 interviewed cancer patients, 54% were classified as fatigued by the P-ICD10 criteria. Internal consistency of P-ICD10 was very good (alpha coefficient 0.82). The principal component analysis corroborated good internal consistency with all variables included in the first component; a second component was used to identify psychological fatigue (concentration and short-term memory disabilities). An abridged set of screening tools based on the first three general symptoms of the P-ICD10 is proposed with 100% specificity and 86% specificity, respectively. There was a marked decrease in FACT-F and VAS1 scores in patients diagnosed as fatigued by the P-ICD10 (mean+/-SD, FACT-F 20+/-9 vs 39+/-8, VAS1 34+/-21 vs 61+/-21). A logistic regression model between P-ICD10 criteria diagnosis and FACT-F (VAS1) identified a score of 34 (61) on the FACT-F scale as a proposed cut-off point for the diagnosis of fatigue. The ICD-10 criteria can be recommended as a diagnostic tool, whereas the FACT-F scale and the level of energy 100-mm VAS assess the intensity of fatigue, and are more suitable for follow-up of cancer-related fatigue. PMID- 15549425 TI - Depression and the use of complementary medicine among breast cancer patients. AB - This was a cross-sectional study to examine the association between anxiety, depression and quality of life and the use of complementary and alternative medicine. Anxiety and depression was measured using the Hospital Anxiety and Depression Scale (HADS), and quality of life was measured using the global quality of life subscale selected from the European Organization for Treatment and Research of Cancer (EORTC) quality of life core questionnaire (QLQ-C30). In all, 177 breast cancer patients were studied, and 32% (n=57) reported that they used or were using complementary medicine. Users and nonusers did not differ significantly in almost all variables studied, with the exception of duration of their diagnosis. The most commonly used complementary medicine was prayer and spiritual healing (n=45, 73.8% of responses). Performing the logistic regression analysis controlling for age, marital status, educational level, knowledge of diagnosis, time since diagnosis, global quality of life, depression, and anxiety scores, the results indicated that the use of complementary medicine among breast cancer patients was associated with sever depression (odds ratio 2.49, 95% CI 1.06-5.89, P 0.04). The other variables studied did not show any significant results. The study findings confirm that the use of complementary medicine is more common among depressed breast cancer patients and might be a marker of greater psychological distress in this group of patients. PMID- 15549426 TI - Delayed emesis: moderately emetogenic chemotherapy. AB - Data on the incidence and efficacy of antiemetic prophylaxis against delayed emesis induced by moderately emetogenic chemotherapy are scanty. An overview of the literature has been done that showed the efficacy of dexamethasone in two of three randomized trials. Its optimal dose and duration of administration has not been defined. Only one of four randomized studies showed a statistically significant efficacy of 5-HT(3) antagonists. Finally, only weak evidence has been published on the efficacy of dopamine receptor antagonists. PMID- 15549427 TI - Current practice and knowledge of oral care for cancer patients: a survey of supportive health care providers. AB - BACKGROUND: The Oral Care Study Section of the Multinational Association of Supportive Care in Cancer (MASCC) and the International Society for Oral Oncology (ISOO) conducted a survey on clinical practices of oral/dental management of cancer patients among supportive health care providers. The main purpose was to evaluate the knowledge and current practice for preventing and managing oral side effects associated with intensive chemotherapy (ICT), hematopoietic cell transplant (HCT), and radiation therapy to the head and neck (H&N RT). MATERIALS AND METHODS: A questionnaire designed and pretested was sent to 212 MASCC/ISOO members around the world with different dental and medical backgrounds. MAIN RESULTS: Seventy-four individuals (35%) responded. The majority of respondents were aware of possible oral complications and provided patients with clinical strategies and recommendations although there was considerable variability among the respondents. Approximately 75% stated that patients were referred for oral/dental care prior to H&N RT and ICT including HCT. However, integrated dental and medical services were reported available in only about 25% of the institutions, and most patients were referred to community-based dental professionals. MAIN CONCLUSIONS: The survey represents a first review of current, international oral care practices. It suggests a need to develop evidence-based clinical guidelines to support effective oral/dental interventions and management strategies for this population. Furthermore, strategies for implementation of oral care protocols and better integration of dental and medical services should be developed. Caution in interpreting these findings is urged due to the limited response rate. PMID- 15549428 TI - Pathogenesis of carcinoma of the papilla of Vater. AB - Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7-, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20-, MUC2-). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa. PMID- 15549429 TI - Hilar cholangiocarcinoma: resectability and radicality after routine diagnostic imaging. AB - BACKGROUND/PURPOSE: En-bloc resection has contributed to the improvement of long term survival in patients with hilar cholangiocarcinoma. In addition, attenuation of intraoperative traumatization of the tumor may decrease tumor spread. The objective of this study was to assess the importance of a routine diagnostic workup for the surgical strategy, radicality, and results in patients with hilar cholangiocarcinoma. METHODS: Between September 1997 and December 2002, 82 patients with hilar cholangiocarcinoma were treated at our department. Preoperative diagnostic workup included endoscopic retrograde cholangiography (ERC), percutaneous transhepatic cholangiography (PTC), computed tomography (CT), and magnetic resonance imaging (MRI). The results of preoperative and retrospective (blinded) assessment of diagnostic data concerning the tumor growth along the bile ducts were compared with the results of surgery. RESULTS: The resection rate was 75%, and the hospital mortality, 7%. The prospective assessment of the resection to be performed was correct in 81% of cases. In ERC, magnetic resonance cholangiography (MRC), and PTC, tumor assessment was precise in 29%, 36%, and 53%, of cases, respectively. Overestimation occurred more frequently than underestimation. The 3-year survival of patients with formally curative or palliative en-bloc resection was 61% and 15%, respectively. For the 9 patients with hilar resection, the 3-year survival was 25%. Survival of patients was comparable, regardless of whether their tumor had been correctly assessed or over- or underestimated. In the multivariate analysis, R0 resection was the only significant prognostic factor ( P = 0.011). CONCLUSIONS: Our routine diagnostic approach led to high resection and survival rates. Obviously a sophisticated diagnostic workup is not an absolute prerequisite for adequate surgery. PMID- 15549430 TI - Increased risk of cholelithiasis after esophagectomy. AB - BACKGROUND/PURPOSE: Truncal vagotomy enhances gallstone formation. As esophagectomy involves truncal vagotomy, it was hypothesized that esophagectomy would increase the risk of cholelithiasis. This study was intended to test this hypothesis and to elucidate factors influencing the incidence of cholelithiasis after esophagectomy. METHODS: The study was a retrospective analysis of 136 patients with esophageal carcinoma who had survived for 5 years or longer after esophagectomy. Eight patients (5.9%) had cholelithiasis before esophagectomy. Of the remaining 128 patients, 113 underwent abdominal ultrasonographic examination for cholelithiasis twice a year after esophagectomy; the median follow-up time was 89.5 months (range, 60-117 months). RESULTS: Gallstones developed in 26 (23%) of the 113 patients undergoing regular ultrasonographic examination. The cumulative incidence of cholelithiasis reached a plateau of 34% at 10 years after esophagectomy. Reduction of body mass index after esophagectomy was the strongest independent predictor of gallstone formation after esophagectomy ( P = 0.0001, log-rank test; P = 0.0003, Cox's proportional hazards model). The prevalence of cholelithiasis at 5 years after esophagectomy (18/113; 16%) was significantly higher than that before esophagectomy (8/136; 5.9%; P = 0.012, Fisher's exact test). CONCLUSIONS: Esophagectomy yields an increased risk of the development of cholelithiasis. Truncal vagotomy and postsurgical malnutrition may contribute to this increased gallstone formation after esophagectomy. PMID- 15549431 TI - Intrahepatic rupture of a caudate lobe adenoma in liver adenomatosis. AB - Hepatic adenomatosis is a rare benign disease that is more common in young and middle-aged women who are non-steroid dependent; it is usually symptomatic, progressive, and susceptible to hemorrhagic complications. Malignant transformation within adenomas is rare. The management of hepatic adenomatosis remains difficult due to the absence of predictive signs of complications, other than the size of the adenomas. Resective surgery is usually indicated, but liver transplantation could be an indication in highly symptomatic or progressive forms of the disease and represents the treatment of choice in cases of malignant transformation. We report a case of intrahepatic rupture of a caudate lobe adenoma which occurred in an adolescent with hepatic adenomatosis; we also present a brief review of the literature. PMID- 15549432 TI - Subcapsular hematoma after right-lobe living-donor liver transplantation. AB - Because right-lobe living-donor liver transplantation was introduced in adult-to adult liver transplantation to mitigate the problems of small-for-size grafts, some technical controversies have been reported. This report describes a case of graft subcapsular hematoma due to parenchymal injury. A 53-year-old woman underwent a right-lobe living-donor liver transplantation for acute-on-chronic liver failure due to primary biliary cirrhosis. A huge subcapsular hematoma was discovered by routine Doppler echogram examination on the first posttransplantation day. Relaparotomy findings revealed that rotation of the graft for the hemostasis procedure during the transplant operation had induced a compression injury to the graft by the xiphoid process. It was speculated that a small laceration in the graft parenchyma led to the major subcapsular hematoma. This experience suggests that the graft liver must be handled with special care to prevent potential mechanical injury. PMID- 15549433 TI - Extrapulmonary small-cell carcinoma of the liver. AB - A 53-year-old man was admitted to our hospital for the evaluation of a mass (13 x 10 cm) in the left lobe of the liver seen by imaging studies. On subsequent biopsy of the mass, the lesion was histologically diagnosed as malignant small round-cell tumor, consistent with metastatic small-cell carcinoma. Segment IV segmentectomy was performed. On pathological examination, the mass showed a yellowish-gray granular appearance with multifocal hemorrhage and necrosis. The phenotypes shown by immunohistochemistry revealed characteristic patterns of small-cell carcinoma (neuron-specific enolase [NSE]+, synaptophysin+, c-Kit+, cluster designation [CD]56+, epithelial membrane antigen [EMA]+, cytokeratin [CK]7-). High resolution-computed tomography (HRCT) revealed inactive pulmonary tuberculosis with small calcified tuberculoma in the right upper lobe. Sputum cytology was negative for malignancy. The postoperative course was uneventful, and platinum-based chemotherapy (cisplatin, etoposide) was initiated. PMID- 15549434 TI - Portal and mesenteric vein thrombosis after portal vein embolization in a patient with protein S deficiency. AB - Portal vein embolization can be performed safely, and so far no major complications have been reported. We report an extremely rare complication of portal vein embolization, a case of portal and mesenteric thrombosis in a 65-year old patient with protein S deficiency. Right portal vein embolization was carried out prior to extended right hepatectomy for advanced gallbladder carcinoma involving the hepatic hilus. Computed tomography 14 days after embolization revealed massive thrombosis of the portal and the superior mesenteric veins. A protein S deficiency was found by means of an extensive workup for hypercoagulable state. Portal vein embolization may have triggered a cascade of events that was expressed as portal and mesenteric vein thrombosis resulting from deficiency of protein S. It may be better to determine the concentrations of such coagulation regulators prior to portal vein embolization. PMID- 15549435 TI - Segmental adenomyomatosis of the gallbladder predisposes to cholecystolithiasis. AB - BACKGROUND/PURPOSE: The aim of the present study was to clarify the association between adenomyomatosis of the gallbladder and cholecystolithiasis. METHODS: A cholecystectomy was performed for cholelithiasis or various other conditions in 1099 patients, of whom 608 had cholecystolithiasis. Adenomyomatosis of the gallbladder was classified as one of three variants: segmental, fundal, and diffuse. Segmental adenomyomatosis has an annular stricture dividing the gallbladder lumen into the "neck compartment" and the "fundal compartment". Bile lipid analysis was performed in 8 patients with segmental adenomyomatosis. RESULTS: Adenomyomatosis of the gallbladder was observed in 156 patients (14.2%), of whom 99 had segmental adenomyomatosis, 54 had fundal adenomyomatosis, and 3 had diffuse adenomyomatosis. The prevalence of cholecystolithiasis was higher in patients with segmental adenomyomatosis (88.9%) than in those without adenomyomatosis (52.3%; P < 0.001). Gallstones were detected earlier in patients with segmental adenomyomatosis than in those without ( P < 0.001) and were located predominantly in the fundal compartment. Bile in the fundal compartment had lower concentrations of total bile acids ( P = 0.012), with an increased cholesterol saturation index ( P = 0.012), compared to bile in the neck compartment. CONCLUSIONS: Segmental adenomyomatosis is a condition predisposing to cholecystolithiasis, probably due to the lithogenic environment in the fundal compartment. Fundal or diffuse adenomyomatosis appears to be unrelated to cholecystolithiasis. PMID- 15549436 TI - Hepatic resection for colorectal metastases in the caudate lobe of the liver. AB - Although hepatic resections for colorectal metastases have become established procedures, there is still only a small number of reports of hepatic resections for such metastases in the caudate lobe. From 1993 to 2001, seven patients underwent eight hepatic resections for colorectal metastases in the caudate lobe at our department. The patients were five men and two women, and their ages were from 53 to 73 years. The ratio of synchronous to metachronous liver metastases was 2 : 5. Solitary metastasis was observed in one patient. One patient with a metastasis in the Spiegel lobe and three patients with metastasis in the caudate process underwent partial resection of the site. The other patients underwent resection of the Spiegel lobe (two times), resection of the right-sided caudate lobe, and total caudate lobe resection. The mean (+/-SE) operative time was 315.9 +/- 30.6 min. Mean intraoperative blood loss was 1325.9 +/- 421.1 ml, and mean postoperative hospital stay was 21 +/- 3.7 days. One patient, who underwent sigmoidectomy and hepatectomy as an emergency operation due to ileus, experienced wound infection. No patient died within 12 months after the surgery. Five patients were alive at 24 months, and three at 36 months. The outcome of these patients encourages us to continue performing hepatic resection for colorectal metastases in the caudate lobe, as it is assumed to be a safe and effective procedure. PMID- 15549437 TI - Primary hydatid disease of the gallbladder: a rare clinical entity. AB - Hydatid disease is endemic in Greece, and has been known from Hippocrates' time to cause cysts in the liver. We report here three very rare cases of primary gallbladder hydatid disease without prior history or evidence of concurrent disease activity in any other location. To our knowledge, only two previous reports exist, each of one patient suffering from primary gallbladder hydatid disease. Unlike the insidious hydatid cysts of the liver, gallbladder hydatidosis in our patients was associated with early diagnosis, due to gallbladder dysfunction symptoms presenting early in its course. Moreover, the size of gallbladder cysts compared to liver cysts at diagnosis was small (maximal diameter, <5 cm), making total cyst excision along with cholecystectomy feasible for all of our patients. During long-term follow up of the patients (up to 10 years), no disease recurrences were noted. We provide supporting evidence that primary gallbladder hydatidosis presents a different pathophysiological and clinical course, having better prognosis, when compared with the usual liver disease. PMID- 15549438 TI - Unexpected gallbladder cancer during laparoscopic cholecystectomy. AB - BACKGROUND/PURPOSE: In the present study, we investigated the effectiveness of surgeons in determining incidental gallbladder pathologies at laparoscopic cholecystectomy (LC). METHODS: This study included 548 patients with gallstones who underwent LC between May 1, 2001 and October 15, 2003. The surgeon made an incision on the gallbladder wall for inspection, and palpated the mucosa after removing the gallbladder from the abdominal cavity to look for unsuspected pathologies. If an abnormal mucosa was observed or palpated, it was marked with a silk suture and then histopathologic examination was performed. RESULTS: Fifty of 548 LC specimens were found to be suspi-cious by the surgeon. Histopathological examination of frozen sections revealed incidental pathologies in 15 of these specimens. Strikingly, 5 of these specimens were considered to have gallbladder cancer (GBC). The other incidental pathologies were consistent with adenomyomatosis, xanthogranulomatous cholecystitis, and fibroepithelial and hyperplastic polyps. Four of the other 498 specimens revealed incidental pathologies at definitive histopathological examination, and all of them were consistent with gastric metaplasia. The sensitivity and specificity of the procedure was 78.9% and 93%, respectively. CONCLUSIONS: A simple prosedure; that is, incision and inspection, and palpation of the gallbladder, seems to be useful for the diagnosis of incidental gallbladder pathologies. PMID- 15549439 TI - Right hepatectomy combined with retrohepatic caval resection, using a left renal vein patch graft for advanced cholangiocarcinoma. AB - Combined resection of the inferior vena cava for hepatobiliary malignancies remains a technical challenge. We successfully resected an intrahepatic cholangiocarcinoma involving the retrohepatic vena cava, and reconstructed the caval defect using a left renal vein patch graft. The patient was a 79-year-old man. Preoperative ultrasonography and computed tomography revealed that the tumor was located in the right lobe of the liver and was about 6 cm in diameter. Arteriogram revealed encasement of the right arterial and portal branches. Magnetic resonance imaging scan revealed that the tumor involved the retrohepatic vena cava. The patient underwent a right hepatectomy combined with resection of the retrohepatic vena cava. The resected portion of the caval wall was 3.6 cm long and 2.7 cm wide. The caval defect was reconstructed using a left renal vein patch graft of a rhomboid shape, which was made by oblique incision of the vein graft. The postoperative course was uneventful. Postoperative cavogram showed adequate patency of the reconstructed retrohepatic vena cava. The patient was disease-free 22 months after surgery. In conclusion, major liver resection combined with caval resection and reconstruction can be performed safely. Furthermore, a left renal vein graft can provide a flexible patch according to the form and size of the caval defect. PMID- 15549440 TI - Intraductal papillary-mucinous adenoma developed in the ventral pancreas in a patient with pancreas divisum. AB - A 34-year-old man was admitted to our hospital with the chief complaints of back pain and epigastralgia. The physical examinations on admission disclosed no abdominal tumor. The serum concentration of total bilirubin was 1.4 mg/dl. The serum elastase-1 level was elevated to 526 ng/dl. Computed tomography showed a cystic lesion, 1 cm in diameter, in the head of the pancreas, without dilatation of the main pancreatic duct. Endoscopic retrograde cholangiopancreatography via the papilla of Vater and the accessory papilla revealed an enlarged ventral pancreatic duct and pancreas divisum. The preoperative diagnosis was mucin producing pancreatic tumor in the ventral pancreas of a patient with pancreas divisum. A pylorus-preserving pancreatoduodenectomy was performed. The gross findings of the cut surface of the resected specimen disclosed mural nodules in the dilated duct of the ventral pancreas. A histological examination of the mural nodules in the ventral pancreas revealed mucin and intraductal papillary adenoma. Benign tumors associated with pancreas divisum are rare; to the best of our knowledge, only three cases have been reported. Although in these three patients the tumor developed in the dorsal pancreas, the tumor developed in the ventral pancreas in our patient. PMID- 15549441 TI - Idiopathic extensive peliosis hepatis treated with liver transplantation. AB - A 50-year-old Danish man, who neither had wasting disease nor was taking steroid containing drugs, complained of abdominal distension, due to a markedly enlarged liver. Percutaneous needle biopsies were taken from the liver, and the findings gave suspicion of a neoplastic tumor. Because of reduced liver function and treatment-resistant ascites, he underwent liver transplantation without a definite preoperative diagnosis. The resected liver weighed 2900 g, and almost all of the parenchyma was destroyed and replaced by multicystic blood-filled spaces, diagnosed as extensive peliosis hepatis complicating liver cirrhosis. Extensive peliosis with liver cirrhosis is a rare condition. Only two cases, caused by contraceptives and treated by liver transplantation, are reported in the English-language literature. We could find no cause other than alcohol abuse lasting several years in this patient, and classified the present case as idiopathic extensive peliosis hepatis. Although scarce subjective findings and misleading liver biopsies made an exact diagnosis difficult, an orthotopic liver transplantation was the only treatment for such complicated peliosis hepatis. PMID- 15549442 TI - The honeycomb gallbladder: a new category of acquired pseudo-multiseptate gallbladder. AB - We report two cases of honeycomb gallbladder as a new category of acquired pseudo multiseptate gallbladder associated with chronic cholecystitis with stones. The two patients were elderly women without any abdominal symptoms or abnormality of laboratory data. On the imaging examinations, a hyperechoic collection with acoustic shadowing on the inferior surface of the liver was typically observed on ultrasonography, with multiloculated gallbladder being observed on computed tomography. Macroscopic findings of the cut plane of the gallbladder showed a characteristic appearance, with thin pseudo-septations arising from the wall and bridging the lumen from side to side, with a honeycomb appearance, including small stones. Microscopic findings suggest that these septational structures could have developed over a chronic inflammatory course after acute obstruction of the cystic duct. Cholecystectomy should be the choice of treatment for honeycomb gallbladder from the viewpoint of clinical management. PMID- 15549443 TI - Current status of hepatic resection for hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common tumors worldwide. For this disease, a variety of therapeutic measures have been applied, including hepatic resections, total hepatectomy followed by allografting, transarterial chemoembolization, and percutaneous tumor ablative therapy by ethanol, microwave coagulation, and radiofrequency ablation. This article focuses on the current status of hepatic resections for HCC. PMID- 15549444 TI - Efficacy of metronidazole for the treatment of clarithromycin-resistant Helicobacter pylori infection in a Japanese population. AB - BACKGROUND: Eradication of Helicobacter pylori has become a common treatment for several diseases. There is an increase in antibiotic-resistant strains, which causes the failure of eradication. The aim of this study was to investigate the usefulness of metronidazole for the treatment of H. pylori infection in patients who failed eradication therapy. METHODS: Seventy H. pylori-positive patients who had failed eradication treatment with first-line triple therapy, which consisted of a proton pump inhibitor, amoxicillin, and clarithromycin, were enrolled into the study. Before the second-line therapy, patients underwent endoscopy to obtain H. pylori strains to test susceptibility to antibiotics. Lansoprazole (30 mg b.d.), amoxicillin (750 mg b.d.), and metronidazole (250 mg b.d.) were administered for 1 week, and the result was tested by 13C-UBT. RESULTS: H. pylori was isolated from 62 patients, and 52 of them (83.9%) were clarithromycin resistant. There was no amoxicillin- or metronidazole-resistant strain. No major adverse effects were seen, and all the patients completed the 1-week regimen. The eradication rates of lansoprazole-amoxicillin-metronidazole were 96.2% (51/53; 95% CI, 87.0%-99.5%) using both intention-to-treat analysis and per protocol analysis. CONCLUSIONS: Lansoprazole-amoxicillin-metronidazole triple therapy is an effective and promising second-line H. pylori eradication therapy in a north Japanese population, which has a low frequency of metronidazole resistance. PMID- 15549445 TI - Altered expression of angiogenic factors in the VEGF-Ets-1 cascades in inflammatory bowel disease. AB - BACKGROUND: The VEGF-Ets-1 cascades play important roles in angiogenesis by converting endothelial cells to an angiogenic phenotype. The aim of this study was to clarify whether the VEGF-Ets-1 cascades are involved in the pathogenesis of inflammatory bowel disease (IBD). METHODS: Colonic specimens were taken from 42 patients with ulcerative colitis (UC), 37 with Crohn's disease (CD), 8 with non-IBD colitis, and 21 normal controls. (1) Expression of vascular endothelial growth factor (VEGF), VEGF receptors (Flt-1, KDR), and Ets-1 proteins in colonic mucosa was immunohistochemically examined using specific antibodies. (2) Expression of Ets-1 protein or VEGF, Flt-1, KDR, and Ets-1 mRNA in colonic mucosa was measured by Western blot or RT-PCR. RESULTS: (1) The number of VEGF containing cells was significantly increased in active UC ( P <0.05). The numbers of positive blood vessels (mean +/- SE /mm2) to Flt-1, KDR, and Ets-1 antibodies were significantly increased in active UC (Flt-1: 4.0 +/- 0.84; KDR: 2.4 +/- 0.37; Ets-1: 5.5 +/- 0.77) compared to active CD (Flt-1: 0.6 +/- 0.30; KDR: 0.77 +/- 0.28; Ets-1: 2.0 +/- 0.56) ( P <0.01), non-IBD colitis (Flt-1: 1.0 +/- 0.45; KDR: 1.83 +/- 0.54; Ets-1: 3.0 +/- 1.0), and controls (Flt-1: 0.88 +/- 0.40; KDR: 0.60 +/- 0.22; Ets-1: 1.67 +/- 0.47) ( P <0.01). The numbers of positive cells to these antibodies were also increased in active UC. (2) Expression of Ets-1 protein and Flt-1, KDR, and Ets-1 mRNA was increased in active UC. CONCLUSIONS: Angiogenic factors in the VEGF-Ets-1 cascades were upregulated in UC, but they were relatively downregulated in CD. These alterations might be involved in the pathogenesis of both diseases. PMID- 15549446 TI - In vivo induction of prostasin mRNA in colonic epithelial cells by dietary sodium depletion and aldosterone infusion in rats. AB - BACKGROUND: Parallel induction of prostasin, a novel serine protease, together with epithelial sodium channel (ENaC) in the colon, may be essential for physiological response to increased circulating aldosterone. The aim of the present study was to investigate whether aldosterone induces prostasin mRNA in parallel with enhanced expression of ENaC in colonic epithelial cells. METHODS: Sprague-Dawley rats were maintained on a sodium-depleted diet or subjected to continuous aldosterone infusion up to 4 weeks. Rats were necropsied at 1, 2, or 4 weeks after the beginning of each treatment. Blood was immediately collected and the large intestine was removed. Plasma aldosterone and arginine-vasopressin (AVP) levels were measured by radio-immunoassay. Epithelial cells were isolated from the right and left colon and RNA was extracted. Expression of prostasin and the alpha-, beta-, and gamma-subunits of ENaC was evaluated by quantitative RT PCR or Northern blot analysis. In another series of experiments, T84 cells were stimulated with aldosterone, dexamethasone, and AVP alone or in combination, and prostasin mRNA was measured by quantitative RT-CPR. RESULTS: Treatment with sodium-depleted diet and aldosterone infusion resulted in an increase of plasma aldosterone and induction of prostasin mRNA in the left colon. Expression of three subunits of ENaC also increased in the left colon. Induction of prostasin mRNA was observed when T84 cells were stimulated with corticosteroids plus AVP in vitro. CONCLUSIONS: Aldosterone has a pivotal role for increasing expression of prostasin in epithelial cells of the left colon. AVP may have a synergistic effect on aldosterone-mediated prostasin induction. PMID- 15549447 TI - Evaluation of venous thromboembolism and coagulation-fibrinolysis markers in Japanese patients with inflammatory bowel disease. AB - BACKGROUND: Arterial or venous thromboembolism is rarely encountered clinically as an extradigestive tract complication in inflammatory bowel disease (IBD). However, it is one of the important prognostic factors for IBD patients. The present study was designed to evaluate the relationship between pulmonary embolism (PE) and deep vein thrombosis (DVT) with coagulation-fibrinolysis markers in patients with active IBD. METHODS: In 47 consecutive cases hospitalized due to active IBD [26 with Crohn's disease (CD) and 21 with ulcerative colitis (UC)], we evaluated the disease severity, blood tests, pulmonary ventilation-perfusion scan (V/Q scan), and magnetic resonance venography (MRV) or conventional venography. RESULTS: PE was diagnosed by V/Q scan in 5 (2 with CD and 3 with UC; 10.6%). DVT was diagnosed in 5 (2 with CD and 3 with UC; 10.6%). Of the 47 patients, 8 (17.0%) had venous thromboembolism (either PE or DVT), and 2 of them (4.3%) had both conditions. In UC patients, the thrombosis group was in more severe stages based on endoscopic grading than the nonthrombosis group. In all patients, the thrombosis group were older (50.3 +/- 14.3 years) than the nonthrombosis group (29.2 +/- 11.7 years). Furthermore, the thrombosis group had higher thrombin-antithrombin III complex (13.1 +/- 17.7 ng/ml) and d-dimer (964 +/- 1402 ng/ml) values than the nonthrombosis group (5.3 +/- 5.5 ng/ml, P=0.0245, and 207 +/- 192 ng/ml, P=0.0016, respectively). There were no significant differences in leukocyte and platelet counts, C-reactive protein, and fibrinogen between the two groups. CONCLUSIONS: A high incidence of venous thromboembolism was suggested in Japanese patients with active IBD. We should be careful with thrombosis in treatment of IBD patients, especially those who are of older age and in more severe stages. PMID- 15549448 TI - Partial characterization of cytoprotective mechanisms of lecithin against bile salt-induced bile duct damage. AB - BACKGROUND: We recently demonstrated that cyclosporine A causes a disproportionate reduction of biliary lipid secretion, and this is inhibited by hydrophilic bile salts through the enhancing of biliary lecithin secretion. In the present study, the underlying mechanism of such a cytoprotective action of hydrophilic bile salts was determined with attention to the possible role of lecithin. METHODS: Immortalized mouse cholangiocytes were cultured for 4 h with taurine conjugates of a hydrophobic bile salt (cholate [TC]), and hydrophilic bile salts (ursodeoxycholate [TUDC], betamuricholate [TbetaMC], and alphamuricholate [TalphaMC]), at 200 microM, in the presence or absence of lecithin (5, 10, 25, 50, 100, or 200 microM), followed by flow cytometric detection of apoptosis, using Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining. Cholangiocyte bile salt transporter mRNAs (apical sodium-dependent bile-salt transporter [Asbt] and multidrug resistance protein 3 [Mrp3]) were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Apoptosis was induced by all of the bile salts (TC > TUDC, TbetaMC, and TalphaMC). Interestingly, bile salt-induced apoptosis was inhibited by lecithin in a concentration-dependent manner. Further, RT-PCR showed that the expressions of Asbt and Mrp3 mRNAs were enhanced by all the bile salts, whereas lecithin reduced Asbt expression, but enhanced Mrp3 expression. CONCLUSIONS: These findings indicate that bile salts cause bile-duct cell damage through Asbt-mediated uptake, but that biliary lecithin physiologically inhibits such damage by reducing the expression of this transporter. In addition, the induction of Mrp3 expression by lecithin may play a role in inhibiting the accumulation of bile. Thus, the modulation of lecithin secretion into bile may be another important target for the treatment of biliary disorders. PMID- 15549449 TI - 18F-FDG PET in the detection of extrahepatic metastases from hepatocellular carcinoma. AB - BACKGROUND: Positron emission tomography (PET) with 18F-fluoro-2-deoxy-D-glucose (18F-FDG) is useful in detecting distant metastases from a variety of malignancies. However, its efficiency in detecting distant metastases from hepatocellular carcinoma (HCC) has not been investigated. The aim of this study was to evaluate the usefulness of 18F-FDG PET for the detection of extrahepatic metastases from HCC. METHODS: Nineteen patients suspected of having extrahepatic HCC underwent 18F-FDG PET. Fourteen patients (group A) had extrahepatic lesions, which were detected by conventional studies. In five patients (group B), conventional imaging showed no extra- or intrahepatic lesions, but the tumor marker levels were elevated. The PET results were compared with those obtained by histopathology or by clinical follow-up. RESULTS: The detection rate of 18F-FDG PET was 83% (24 of 29 metastases) for extrahepatic metastases larger than 1 cm in greatest diameter and 13% (1 of 8 metastases) for lesions less than or equal to 1 cm. PET revealed two bone metastases not depicted by bone scan, and detected the nodal metastasis and intestinal metastases inconclusive on computed tomography. Extrahepatic lesions were resected in 5 patients of group A on the basis of PET findings. In all patients of group B, PET results were true negative for extrahepatic metastases, but HCCs were detected in the liver within 4 months in 4 patients. These were no false-positive lesions in either group. CONCLUSIONS: This preliminary study suggested that 18F-FDG PET could provide additional information and contribute to the management of HCC patients suspected of having extrahepatic metastases. PMID- 15549450 TI - Efficacy of 6-month interferon therapy in chronic hepatitis B virus infection in Japan. AB - BACKGROUND: In Japan, there are few studies of long-term (more than 1 month) interferon (IFN) therapy for chronic hepatitis B (CHB). In this retrospective study, we investigated the efficacy and predictors of response to 6-month IFN therapy. METHODS: We analyzed 66 Japanese patients with CHB who were treated with IFN for 6 months. They comprised patients who were hepatitis B e antigen (HBeAg) positive (n=45) and -negative (n=21). One (2%), 8 (12%), and 51 (77%) patients were infected with hepatitis B virus (HBV) genotypes A, B, and C, respectively. Responders in patients positive for HBeAg were defined as those who showed normalization of serum alanine aminotransferase (ALT) level, HBeAg loss, and HBV DNA negativity at 6 months after completion of IFN therapy. In patients negative for HBeAg, responders were defined as those patients who showed normalization of ALT level and HBV DNA negativity at the same 6-month time point. RESULTS: Of the 45 patients with HBeAg at the commencement of IFN therapy, 9 (20%) were responders. Young patients, especially those with a high serum ALT level, were significantly more likely to respond to IFN therapy. Of the 21 patients negative for HBeAg, 13 (62%) were responders. There were no significant differences in clinical characteristics between responders and nonresponders among patients negative for HBeAg. Multivariate analyses identified HBeAg negativity and young age as independent factors associated with a positive response to 6-month IFN therapy. However, long-term follow-up of the treated patients showed a fall in the response rate. CONCLUSIONS: The response rate to 6-month IFN therapy among HBeAg-positive patients was low. However, young patients may require long-term IFN therapy. PMID- 15549451 TI - A comparative clinical study of gallbladder tubular adenocarcinoma according to the grade of differentiation. AB - BACKGROUND: We sought to elucidate differences in the clinical characteristics of gallbladder carcinoma according to the histological degree of glandular formation. METHODS: The subjects consisted of 66 autopsy cases out of 331 patients admitted for gallbladder carcinoma between 1975 and 2000. After identifying 49 tubular adenocarcinoma cases, we arbitrarily divided them into two categories: a well-differentiated type (group W; n=41; well or moderately differentiated tubular adenocarcinoma), and a poorly differentiated type (group P, n=8). The clinical features of the groups were compared. RESULTS: No significant differences were found with regard to sex, age, or serum carcinoembryonic antigen (CEA) levels between the two groups. Moreover, no intergroup difference was found in either the time interval from the initial symptoms to admission, or the grade of tumor progression on admission. Although the frequency of liver metastases on admission was similar in both groups, the frequency at autopsy was significantly higher in group W than in group P (73% vs 25%; P <0.05). When survival was determined based on the duration after primary treatment, the longest and median periods were 48.7 months and 3.7 months, respectively, in group W, whereas none of the subjects in group P survived for over 2 months. This difference was statistically significant ( P <0.001). CONCLUSIONS: Hematogenous metastasis was more prominent in group W. Poorly differentiated tubular adenocarcinoma can be closely associated with a poor prognosis. PMID- 15549452 TI - Biliary excretion of bile acids and organic anions in rats with dichloroethylene induced bile canalicular injury. AB - BACKGROUND: Hepatocytes in zone 1 of the hepatic lobule play a role in the uptake and biliary excretion of bile acids and organic anions under physiological conditions, and hepatocytes in zone 3 may play a role only when there is a high dose load. To further elucidate the role of hepatic zonation in the hepatic handling of bile acids and organic anions, the biliary excretion of these compounds was studied in rats with dichloroethylene (DCE)-induced selective zone 3 bile canalicular injury. METHODS: Biliary excretion of various bile acids and organic anions was studied in rats 1 h after oral administration of DCE (5 mg/100 g). The effect of DCE on the immunostaining of multidrug resistance protein 2 (Mrp2; an important canalicular organic anion transporter) in the liver was also examined. RESULTS: The biliary excretory maximum of taurocholate and tauroursodeoxycholate was decreased in DCE-treated rats, whereas the biliary excretion of taurolithocholate-sulfate and phenolphthalein-glucuronide was unchanged in DCE-treated rats, and DCE treatment decreased the biliary excretion of sulfobromophthalein and pravastatin. DCE decreased Mrp2 staining in the canalicular membrane of zone 3 hepatocytes on immunohistochemistry. CONCLUSIONS: These findings indicate that canalicular transport in zone 3 hepatocytes is important in the biliary excretion of bile acids and organic anions, when they are administered at high doses. PMID- 15549453 TI - Assessment of occult pancreatobiliary reflux in patients with pancreaticobiliary disease by ERCP. AB - BACKGROUND: The aim of this study was to evaluate the degree of occult pancreatobiliary reflux by measuring the biliary amylase levels in the common bile duct (CBDA) and gallbladder (GBA) at endoscopic retrograde cholangiopancreatography (ERCP). METHODS: Eligible patients included 86 consecutive cases of pancreaticobiliary disease with prospective implementation of bile collection during an ERCP procedure. Patients with pancreatobiliary maljunction (PBM) were excluded. Nineteen cases of eligible patients had simultaneous collection of gallbladder bile. Bile was further collected by cholecystectomy in 8 cases. RESULTS: Twenty-two cases (26%) revealed a CBDA level higher than serum amylase (high bile amylase level, HBA group) and 64 cases exhibited a CBDA level lower than serum (LBA group). The mean values of CBDA in the HBA and LBA groups were 5502 IU/l and 29 IU/l, respectively. The rate of HBA was significantly higher in patients who were elderly, had a dilated common bile duct, and those with choledocholithiasis (P <0.05). Three cases (16%) showed a CBDA greater than twice the GBA. Eleven cases (58%) exhibited a GBA higher than the CBDA. The values of GBA obtained during ERCP and cholecystectomy were consistent. CONCLUSIONS: These findings suggest that even non-PBM cases can exhibit occult pancreatobiliary reflux, which can thereby cause biliary disease. PMID- 15549454 TI - Non-multiple lymphomatous polyposis form of mantle cell lymphoma in the gastrointestinal tract. AB - Mantle cell lymphoma (MCL) comprises 2.5%-7% of all non-Hodgkin's lymphomas, and the gastrointestinal tract is involved in about 20% of cases. Multiple lymphomatous polyposis (MLP) is an uncommon disease that is regarded as the intestinal form of MCL. We present a rare case of gastrointestinal MCL without MLP, and demonstrate that rituximab was effective for the treatment of this patient. A 61-year-old man presented with continuous diarrhea and hematochezia for a period of 5 months. Superficial lymph nodes were not palpable, but both tonsilla were enlarged. The level of soluble interleukin (IL)2-receptor was 3480 U/ml (normal <500 U/ml). Colonoscopy showed diffuse redness with erosion, without observation of any venous capillary, with these findings continuing from the rectum to the ileum. Upper gastrointestinal endoscopy showed a slightly rough gastric mucosal surface, and chicken-skin like mucosa was observed in the second portion of the duodenum. Small-to-medium size lymphoma cells were seen histologically from the tonsilla to the rectum. The lymphoma cells were immunohistochemically positive for CD5, CD20, CD79a, and cyclin D1. Polymerase chain reaction analysis revealed a chromosomal translocation t(11;14)(q13;q32) in the bcl-1 gene. We diagnosed this as a case of MCL from these findings. For treatment, the patient received a total of ten courses of combination chemotherapy consisting of cyclophosphamide (1000 mg), doxorubicin (70 mg), vincristine (2 mg) and prednisolone (50 mg) (CHOP), which led to a partial remission. However, 2.5 years later, massive infiltrations of the lymphoma cells were found in the colon and stomach. As the infiltrating lymphoma cells expressed CD20 molecules on their surfaces, the patient was treated with a chimeric anti CD20 monoclohal antibody, rituximab, which showed significant efficacy, and a second partial remission was achieved. PMID- 15549455 TI - Gastrointestinal stromal tumor in the jejunum: diagnosis and control of bleeding with electrocoagulation by using double-balloon enteroscopy. AB - A 43-year-old man presented with gastrointestinal bleeding. A tumor with central ulceration was observed in the jejunum, with the use of a new enteroscopy system called "double-balloon enteroscopy". Bleeding after biopsy sampling of the tumor was controlled endoscopically by using electrocoagulation. Histological findings of the biopsy specimens were consistent with gastrointestinal stromal tumor, and this was surgically resected. Double-balloon enteroscopy was useful for the diagnosis as well as the control of bleeding in this patient. PMID- 15549456 TI - Autoimmune pancreatitis with pseudocysts. AB - A 47-year-old woman was admitted to our hospital with complaints of fever, upper abdominal pain, and back pain. The serum amylase, C-reactive protein (CRP), and IgG (especially IgG4) were elevated, and abdominal computed tomography (CT) revealed diffuse enlargement of the pancreas and pseudocysts. Endoscopic retrograde pancreatography (ERP) revealed diffuse irregular narrowing of the main pancreatic duct. Histopathological examination of the pancreatic tissue showed fibrotic change with lymphocytic infiltration. Based on these findings, we diagnosed this case as a case of autoimmune pancreatitis. This case also fully satisfied the diagnostic criteria for autoimmune pancreatitis established by the Japan Pancreas Society in 2002. Few reports have been published on cases of autoimmune pancreatitis complicated by the formation of pseudocysts in the pancreas. We, therefore, report this case here to emphasize that cases of autoimmune pancreatitis can be complicated by the development of pseudocysts. PMID- 15549457 TI - Periampullary somatostatinoma and multiple gastrointestinal stromal tumors associated with von Recklinghausen's disease. PMID- 15549458 TI - Intrahepatic splenosis in a patient with chronic hepatitis C. PMID- 15549459 TI - Hepatocellular carcinoma that ruptured during radiofrequency ablation therapy. PMID- 15549460 TI - FDG-PET and hepatocellular carcinoma. PMID- 15549461 TI - Evidence-based medicine on domestic data. PMID- 15549462 TI - Pancreatic juice can reflux into the bile duct in patients without anomalous pancreaticobiliary junction. PMID- 15549463 TI - Multiple lymphomatous polyposis form is common but not specific for mantle cell lymphoma in the gastrointestinal tract. PMID- 15549464 TI - Laryngotracheal application of lidocaine spray increases the incidence of postoperative sore throat after total intravenous anesthesia. AB - PURPOSE: To determine the effect of laryngotracheal application of different doses of lidocaine spray on postoperative sore throat and hoarseness, we evaluated the incidence and severity of these complications in 168 ASA I-III patients aged 15-92 years in a placebo-controlled study. METHODS: After induction of anesthesia with propofol, ketamine, fentanyl, and vecuronium, the laryngotracheal area was sprayed immediately before intubation with lidocaine spray either 5 times (L5 group, n = 47) or 10 times (L10 group, n = 48) or with normal saline 1 ml (placebo group, n = 51). Postoperative sore throat and hoarseness were evaluated immediately after surgery and on the day after surgery. RESULTS: The incidence of sore throat was significantly higher in the L10 group than in the placebo group on both the day of and the day after surgery. The severity of sore throat was significantly higher in the L5 and L10 groups than in the placebo group on the day of surgery. On the day after surgery, the severity of sore throat remained significantly higher in the L10 group than in the placebo group. Although the incidence and severity of sore throat increased in a dose dependent manner, these were not significantly different between the L5 and L10 groups. In addition, the incidence and severity of hoarseness did not differ at all among the three groups. CONCLUSION: We recommend that applications of lidocaine spray to the laryngotracheal area should be avoided to help eliminate unnecessary postoperative sore throat, thereby leading to improvement in patient satisfaction. PMID- 15549465 TI - Changes in respiratory pattern during continuous positive airway pressure in infants after cardiac surgery. AB - PURPOSE: Spontaneous breathing trials are commonly used in adults to enable smooth weaning from mechanical ventilation. However, few investigations have examined spontaneous breathing tests in infants. We investigated how respiratory patterns of infants changed during continuous positive airway pressure (CPAP) and whether successful extubation followed CPAP. METHODS: Fifty-one consecutive post cardiac surgery infants satisfied the following weaning criteria: stable hemodynamics, pH > 7.30, tidal volume > 5 ml.kg(-1), and respiratory rate < 50 breaths.min(-1) with pressure control of 10-16 cm H(2)O. We applied CPAP of 3 cm H(2)O for 30 min to these 51 infants. During CPAP, tidal volume, respiratory rate, and arterial blood gases were measured. CPAP was terminated if the patient showed a sustained increase or decrease in heart rate or blood pressure (>20%), a decrease in arterial oxygen saturation (>5%), agitation, or diaphoresis. After the completion of CPAP, tracheal extubation was performed. We considered extubation successful if no reintubation was required in the ensuing 48 h. RESULTS: Although hemodynamic and ventilatory variables were unstable for the first 5 min, they stabilized after 10 min of CPAP. Fifty infants completed the CPAP trial safely. Of these, 46 (92%) underwent successful extubation after the CPAP trial. The failure group (4 infants) showed lower pH, higher arterial carbon dioxide tension, and more rapid shallow breathing during CPAP than the success group. CONCLUSION: After cardiac surgery, when infants recovered stable hemodynamics and spontaneous breathing, the ventilatory pattern and hemodynamics became stable after 10 min of CPAP. Ninety-two percent of the patients were successfully extubated following a 30-min CPAP trial. PMID- 15549466 TI - Ropivacaine produces sensory blockade in the lumbar sacral region more frequently than mepivacaine in lower thoracic epidural anesthesia. AB - PURPOSE: The first sacral nerve has the largest diameter among the spinal nerves and is resistant to local anesthetics. Ropivacaine is a newly developed local anesthetic. There is a possibility that a difference in chemical properties between ropivacaine and other local anesthetics produces a difference in the blockade of the S1 dermatome by lower thoracic epidural anesthesia. Mepivacaine, 2%, is frequently used for epidural anesthesia and produces a level of blockade similar to that of bupivacaine, 0.5%. The purpose of this study was to examine the sensory blockade in the sacral region induced by ropivacaine with that induced by mepivacaine administered in the lower thoracic epidural space. METHODS: Eighteen adults undergoing lower thoracic epidural anesthesia (thoracic 11/12 interspace) were studied in a double-blind fashion. Patients were assigned to one of two groups: those who received 2% mepivacaine, 18 ml (group M; n = 9), and those who received 1% ropivacaine, 12 ml (group R; n = 9). The cephalad levels of sensory blockade to cold, pinprick, and touch in the L2, S1, and S3 dermatomes were assessed at 10, 20, and 35 min after injection. RESULTS: There were no differences in the cephalad levels of sensory block to cold (T4 [range, T4-T2] and T4 [range, T6-T2]), pinprick (T4 [range, T6-T4] and T4 [range, T6 T4]), or touch (T6 [range, T6-T4] and T6 [range, T6-T4]) between group M and group R respectively, at 35 min (P > 0.05). In the L2 and S3 dermatomes, there were no significant differences in the numbers of patients who obtained sensory block to cold or pinprick at 20 and 35 min after study drug administration. However, in the S1 dermatome, significantly higher numbers of patients in group R obtained sensory block to cold at 20 and 35 min after study drug administration than in group M (8 and 0; 9 and 0; P = 0.001 and P < 0.001; 20 min and 35 min after administration, respectively). Also in the S1 dermatome, significantly higher numbers of patients in group R obtained sensory block to pinprick at 20 and 35 min after study drug administration than in group M (6 and 0; 9 and 0; P = 0.027 and P < 0.001; 20 min and 35 min after administration, respectively). A significantly higher number of patients in group R had sensory block to touch in the S3 dermatome at 35 min (7 and 2; group R and group M, respectively; P = 0.01). CONCLUSION: Ropivacaine, 1%, administered in the lower thoracic epidural space, induces sensory blockade to cold and pinprick in the S1 dermatome more frequently than 2% mepivacaine. PMID- 15549467 TI - Dose-finding study of intravenous midazolam for sedation and amnesia during spinal anesthesia in patients premedicated with intramuscular midazolam. AB - PURPOSE: We investigated the effective and safe dose of intravenous midazolam for sedation and amnesia during spinal anesthesia in patients premedicated with intramuscular midazolam. METHODS: One hundred and eighty patients aged 20-50 years scheduled for spinal anesthesia received midazolam 0.06 mg.kg(-1) and atropine 0.01 mg.kg(-1) intramuscularly 15 min before entering the operating room. Spinal anesthesia was performed with 0.5% hyperbaric tetracaine. Five minutes after starting surgery, midazolam 0 (control group), 0.01, 0.02, 0.03, 0.04, or 0.05 mg.kg(-1) was intravenously administered (30 patients each). Blood pressure, heart rate, respiratory rate, percutaneous oxygen saturation (S(p)(O)(2)), verbal response, eyelash reflex, and involuntary body movement were measured every 5 min for 30 min. Memory during surgery was also investigated. RESULTS: The number of the patients with loss of verbal response, with loss of eyelash reflex, and with no memory during surgery were significantly larger in the groups receiving midazolam >or=0.03 mg.kg(-1), >or=0.04 mg.kg(-1), and >or=0.02 mg.kg(-1), respectively. The decrease in blood pressure or increase in respiratory rate with decrease in S(p)(O)(2) was significantly larger in the groups receiving midazolam >or=0.03 mg.kg(-1) or 0.05 mg.kg(-1), respectively. CONCLUSION: For sedation and amnesia of the patients aged 20-50 years in spinal anesthesia with about 1 h duration receiving intramuscular midazolam 0.06 mg.kg( 1) as a premedication, intravenous midazolam 0.02 mg.kg(-1) might be effective and safe. PMID- 15549468 TI - Comparison between a disposable and an electronic PCA device for labor epidural analgesia. AB - PURPOSE: The aims of the present study were (1) to investigate if a disposable patient-controlled analgesia (PCA) device can be used for labor analgesia and (2) to evaluate the device by midwives and parturients. METHODS: Forty healthy parturients were divided into two groups and received combined spinal epidural analgesia for labor pain relief. Following intrathecal administration of 3 mg ropivacaine and 1.5 microg sufentanil, either a disposable PCA device (Coopdech Syrinjector; Daiken Medical, Osaka, Japan) or an electronic PCA device (IVAC PCAM PCA Syringe Pump; Alaris, Basingstoke, UK) was connected to the epidural catheter, and 0.15% ropivacaine with sufentanil 0.75 microg/ml was used for continuous infusion and PCA. For an electronic PCA device, continuous infusion rate, bolus dose, lockout time, and hourly limit were set at 4 ml/h, 3 ml, 15 min, and 16 ml, respectively. For a disposable PCA device, continuous infusion rate, bolus dose, and an hourly limit were set at 4 ml/h, 3 ml, and 16 ml, respectively, but lockout function was not available. RESULTS: No differences were observed between the groups concerning demographic data, obstetric data, and outcome of labor. Anesthetic requirements (disposable, 9.7 +/- 4.7 ml/h; electronic, 8.2 +/- 4.0 ml/h) and VAS score during the delivery (disposable, 26 +/- 25; electronic, 21 +/- 22) were similar between the groups. Midwives praised the disposable PCA device as well as the electronic one. CONCLUSION: The present results imply that the disposable PCA device can be an alternative to the electronic PCA device for labor analgesia. PMID- 15549469 TI - Mapping of putative ether-anesthesia resistance gene using C57BL/6J and MSM/Ms mouse strains. AB - PURPOSE: We attempted to identify the locations of major mouse genes responsible for sensitivity to diethylether (ether) anesthesia, using microsatellite linkage analyses including Quantitative Trait Locus (QTL) analysis. METHODS: To determine the locations of ether anesthesia resistance genes on chromosomes, an ether anesthesia-resistant mouse strain, C57BL/6J (C57BL), and an ether anesthesia sensitive mouse strain, MSM/Ms (MSM), were used. The sensitivity of mice to ether anesthesia was determined from the latency time required to lose the righting reflex during exposure to 4% ether vapor in air. The (C57BL x MSM) F(1) mice were found to be resistant to ether, showing that the resistant phenotype is genetically dominant. Twelve resistant and 12 sensitive mice were then selected from the 196 backcrossed F(2) mice (F(1) x MSM) at 11-16 weeks of age. Genomic DNA samples were extracted from the tails for mapping ether anesthesia-related genes using microsatellite linkage analyses. RESULTS: One major putative gene related to resistance to ether anesthesia was restricted in the region 23 to 37 cM from the centromere in chromosome 7 by primary and secondary linkage analyses. The QTL analysis narrowed the position of the gene to 29.0 cM, with a maximum logarithm of odds (LOD) score of 3.03, and it was termed Etan1 ( ether-anesthesia 1). CONCLUSION: Microsatellite linkage analyses, including QTL analysis, determined the location of the ether-resistance gene, Etan1, within a narrow range. Our findings should be helpful for further experiments, such as cloning of the gene governing the sensitivity to ether anesthesia in mice. PMID- 15549470 TI - Amsorb Plus and Dragersorb Free, two new-generation carbon dioxide absorbents that produce a low compound A concentration while providing sufficient CO2 absorption capacity in simulated sevoflurane anesthesia. AB - PURPOSE: The properties of two new-generation CO(2) absorbents, Amsorb Plus (Armstrong Medical, Coleraine, UK) and Dragersorb Free (Drager, Lubeck, Germany), were compared with those of Amsorb (Armstrong Medical) and Sodasorb II (W.R. Grace, Lexington, MA, USA). METHODS: The concentration of compound A produced by each absorbent was determined in a low-flow circuit containing sevoflurane, and the CO(2) absorption capacity of the absorbent was measured. The circuit contained 1000 g of each absorbent and had a fresh gas (O(2)) flow rate of 1 l.min(-1) containing 2% sevoflurane. CO(2) was delivered to the circuit at a flow rate of 200 ml.min(-1). RESULTS: The maximum concentrations of compound A were 2.2 +/- 0.0, 2.3 +/- 0.3, 2.2 +/- 0.2, and 23.5 +/- 1.5 ppm (mean +/- SD) for Amsorb Plus, Dragersorb Free, Amsorb, and Sodasorb II, respectively. The maximum concentration of compound A for Sodasorb II was significantly higher than those for the other absorbents (P < 0.01). The CO(2) absorption capacities (time taken to reach an inspiratory CO(2) level of 2 mmHg) were 1023 +/- 48, 1074 +/- 36, 767 +/- 41, and 1084 +/- 54 min, respectively, and the capacity of Amsorb was significantly lower than that of the other absorbents (P < 0.01). CONCLUSION: The new-generation carbon dioxide absorbents, Amsorb Plus and Dragersorb Free, produce a low concentration of compound A in the circuit while showing sufficient CO(2) absorption capacity. PMID- 15549471 TI - A rabbit model for evaluation of surgical anesthesia and analgesia: characterization and validation with isoflurane anesthesia and fentanyl analgesia. AB - PURPOSE: With a clamp test, quantitative estimation of the level of surgical anesthesia/analgesia is not easy. We have developed a rabbit pain model allowing for quantitative evaluation of the level of surgical anesthesia/analgesia using both electrical and mechanical stimuli as simulated surgical stimuli. We evaluated whether this model allows for accurate tracing of dynamically changing levels of surgical anesthesia/analgesia induced by isoflurane and fentanyl. METHODS: Eight rabbits tracheotomized and vascularly cannulated under 3% isoflurane anesthesia were placed on a sling that allowed for free movement of the head and extremities. After the isoflurane concentration was reduced stepwise to 1.5% and then to 0%, four graded doses of fentanyl (5, 10, 20, and 40 microg.kg(-1)) were injected intravenously at intervals of 120 min. At each dose, anesthetic/analgesic end-point variables were determined, including the number of animals behaviorally unresponsive to clamping the forepaw (nonresponders) and the threshold voltage of subcutaneous electrical stimulation (2, 5, and 50 Hz) required to evoke the head lift (HLT: pain detection/arousal threshold: sedative/hypnotic index) and the escape movement (EMT: pain tolerance threshold: analgesic index). RESULTS: With increasing doses of isoflurane and fentayl, HLTs and EMTs, especially those at 5 Hz, increased dose-dependently and proportionately to increases in the number of nonresponders to clamping the forepaw, a standard indicator of the anesthetic/analgesic level. CONCLUSION: Using the HLT and EMT, especially at 5 Hz, combined with a clamp test, this rabbit model allows for repeated, quantitative, and distinctive evaluation of the dynamically changing levels of both sedative/hypnotic and analgesic components of surgical anesthesia/analgesia. PMID- 15549472 TI - Long-term pain relief effects in four patients undergoing percutaneous vertebroplasty for metastatic vertebral tumor. AB - We reviewed long-term pain relief in four patients undergoing percutaneous vertebroplasty (PVP) for lumbar or back pain due to metastatic vertebral tumors. The patients received anesthesiological palliative care with analgesics until their death after PVP. Pain intensity, the presence or absence of recurrence of pain, changes in dosage of analgesics given before and after PVP, and complications associated with PVP were evaluated. A numerical rating scale (NRS) from 0 (no pain) to 10 (worst pain imaginable) was used to measure pain. The patients were three men and one woman (mean age, 58 years). PVP was performed in five vertebrae (one thoracic and four lumbar). The NRS scores on moving before PVP were 10, 8, 10, and 10. After PVP, NRS decreased to 0, 3, 5, and 0, respectively, within 72 h. No recurrence of pain in the treated area occurred until death in any of these patients. The dosages of analgesics given were decreased in two cases, but no changes were made in the other two cases. There were no complications associated with PVP. Percutaneous vertebro-plasty is a safe and effective treatment for long-time pain relief in patients with metastatic vertebral tumors. PMID- 15549473 TI - Erythemas caused by electrodes while monitoring neuromuscular blockade: three cases. AB - Skin erythemas formed in three patients during surgery at the sites where negative electrodes had been attached to stimulate the ulnar nerve for a neuromuscular transmission monitor (Relaxograph). The patients were all women, aged 52, 62, and 74 years, and general anesthesia lasted 8 h 20 min, 4 h 50 min, and 8 h 45 min, respectively. The electrodes used were disposable ECG electrodes in the first two patients and one designed for a neuromuscular monitor in the third; all were carbon-coated and then covered with gel. However, when the electrodes were detached from the lesion, they all showed loss or damage of the carbon coating under the gel. We recommend balancing the merit of monitoring with the risk of complications, even when applying an apparently safe, noninvasive monitor. PMID- 15549474 TI - Continuous spinal anesthesia and postoperative analgesia for elective cesarean section in a parturient with Eisenmenger's syndrome. AB - We describe the use of continuous spinal anesthesia (CSA) for an elective cesarean section in a 29-year-old parturient with Eisenmenger's syndrome at 30 weeks of gestation. It is essential in patients with Eisenmenger's syndrome to prevent significant increases in right-to-left shunt following the reduction in systemic vascular resistance. In this case, the patient hoped to be awake during the operation because of her fear of death. We therefore applied CSA to this patient because single-shot spinal anesthesia and epidural anesthesia might cause sudden cardiovascular depression. In fact, sudden cardiovascular changes were avoided by the titration of local anesthetics and the operation was uneventful, although prompt treatment of hypotension was essential and adjustment of the anesthetic levels was difficult. Postoperative patient-controlled spinal analgesia provided satisfactory pain relief with hemodynamic stability and no significant side effects. However, thorough experience with the requisite techniques is critical in CSA because of the technical difficulty of the procedure, and anesthesiologists must gain such experience in less-demanding cases before attempting to administer it in patients presenting extreme challenges as described in this case report. PMID- 15549475 TI - Conversion of atrial fibrillation to sinus rhythm during landiolol infusion. AB - A 71-year-old woman with a history of persistent atrial fibrillation underwent clipping of a ruptured cerebral artery aneurysm. During the surgery her cardiac rhythm was atrial fibrillation and the ventricular rate increased to 130 beats.min(-1). Administration of landiolol was started with 1-min loading infusion at 0.125 mg.kg(-1).min(-1) and continuous infusion at 0.04 mg.kg( 1).min(-1), which was effective in controlling the ventricular rate without causing hypotension. Approximately 120 min after the landiolol infusion was started, the atrial fibrillation was converted to sinus rhythm. Her sinus rhythm was maintained until she left the operating room, even after discontinuation of landiolol. PMID- 15549476 TI - Unanticipated hemothorax during general anesthesia. AB - We experienced a case of uncommon spontaneous hemothorax during general anesthesia. A 72-year-old woman underwent emergency repair of damaged ascending colon. Her chest roentgenogram before surgery revealed no abnormalities. After the uneventful surgery, the trachea was extubated and the patient breathed without difficulty with stable vital signs. However, the S(p)(O)(2) varied around 95% with a 100% oxygen mask. Chest roentgenogram obtained after extubation showed a massive amount of fluid in the pleural cavity. A chest drainage tube was inserted, and approximately 1000 ml of frank blood was drawn without air leakage. Thereafter, her oxygenation remarkably improved. We could not determine the etiology of the hemothorax. PMID- 15549477 TI - A case of complex regional pain syndrome type II after transradial coronary intervention. AB - The transradial approach for coronary catheterization is now a routine technique without serious complications at the puncture site. We report a case of complex regional pain syndrome type II (CRPS type II) in the hand after the transradial coronary intervention, which may alert medical personnel that the technique may cause serious regional pain with disability. A 61-year-old woman underwent coronary intervention via the right radial artery for the treatment of unstable angina. After the operation she complained of severe pain in the right hand, consistently felt along the median nerve distribution. The nerve conduction study suggested carpal tunnel syndrome. We made a diagnosis of CRPS type II, and the patient received stellate ganglion blockade, cervical epidural blockade, and administration of amitriptyline and loxoprofen. The symptoms gradually improved and her activities of daily living markedly improved. The median nerve appeared to be damaged by local compression and potential ischemia. Careful attention should be paid to avoid CRPS type II, associated with excess compression. PMID- 15549478 TI - Expiratory tidal volume displayed on Bird 8400 STi can exceed the preset tidal volume due to cardiogenic oscillation: a lung model study. AB - We noticed that monitored tidal volumes often exceeded preset tidal volumes in patients with large cardiogenic oscillation. To investigate whether triggering modes affect this discrepancy, we simulated cardiogenic oscillation of 90 breaths/min in a lung model, which was ventilated with a Bird 8400 STi ventilator (Bird, Palm Springs, CA, USA). The magnitude of cardiogenic oscillation was defined as peak expiratory flow fluctuation at the lung model. Two respiratory rates (5 and 10 breaths/min) and two triggering modes (flow-triggering and pressure-triggering) were applied, while tidal volume was set at 500 ml. We recorded tidal volume on a ventilator monitor and calculated the discrepancy from the set tidal volume. We also measured fluctuation in flow and airway opening pressure created by cardiogenic oscillation. During flow-triggering, larger flow fluctuation and smaller airway pressure fluctuation were observed compared with during pressure-triggering. During flow-triggering, the discrepancy between monitored tidal volume and set value ranged from 0 to +327 ml at 5 breaths/min, and from 0 to +105 ml at 10 breaths/min. There was a linear correlation between the magnitude of cardiogenic oscillation and the overestimation of tidal volume. By contrast, during pressure-triggering, the discrepancy was small. In conclusion, tidal volume is overestimated during flow-triggering but not during pressure-triggering when cardiogenic oscillation is large. PMID- 15549479 TI - Performance of a tonometer for arterial pressure measurement during anesthesia in the elderly. AB - We investigated performance of an arterial tonometer during anesthesia in the elderly. Thirty patients (17 men and 13 women), ranging from 70 to 86 years old, were included, and agreement between tonometric arterial pressure (TAP) and intraarterial pressure (IAP) was calculated. A total of 6487 paired points was sampled, and values for precision (mean absolute difference +/- SD) were 6.8 +/- 5.2 for systolic, 6.9 +/- 4.6 for mean, and 9.2 +/- 5.4 mmHg for diastolic blood pressures. Values for bias and limits of agreement [mean difference (TAP - IAP) +/- SD, mean difference (TAP - IAP) +/- 2 SD, respectively] for systolic, mean, and diastolic pressure were 1.2 +/- 8.4 and 1.2 +/- 16.8, 5.7 +/- 5.9 and 5.7 +/- 11.8, and 8.6 +/- 6.2 and 8.6 +/- 12.4 mmHg, respectively. Compared with previous data, aging is likely to affect the performance of an arterial tonometer. PMID- 15549480 TI - Intraoperative pneumothrax during retroperitoneal laparoscopic surgery. PMID- 15549481 TI - Re-publication of a translation of 'The vegetative organs of Monotropa hypopitys L.' published by F. Kamienski in 1882, with an update on Monotropa mycorrhizas. AB - This paper begins with a brief comparison of Franz Kamienski's 1882 view of the fungus-root associations and nutrition of Monotropa hypopitys with our current understanding. The rest of this paper is a re-publication of Shannon Berch's 1985 translation of Kamienski's breakthrough paper in which it was asserted that Monotropa forms a mutualistic symbiosis and is nourished by fungi associated with the roots of neighbouring trees. PMID- 15549482 TI - On the nutritional dependence of certain trees on root symbiosis with belowground fungi (an English translation of A.B. Frank's classic paper of 1885). PMID- 15549483 TI - "Does size matter?"A comparison of balloon-assisted less-invasive vs conventional retroperitoneal approach for anterior lumbar interbody fusion. AB - This is a case-series comparison of two approaches to anterior lumbar interbody fusion. A conventional open approach (COA) was compared with a balloon-assisted minimally invasive approach (BMI). Outcome measures included operating time, blood loss and complications. Secondary outcome measures included analgesia requirements, time to mobilization and inpatient stay. There were 17 females (7 COA, 10 BMI) and 18 males (9 COA and 9 BMI). Forty-five discs (21 COA, 24 BMI) in total were fused in 35 patients. There were significant differences (in favour of the BMI) in the overall operating time between the COA and the BMI, and the single level COA and the BMI. There was no inter-group difference in the PCA requirements either overall or between one or two-level operations. The less invasive approach did have a benefit in earlier mobilization of the single-level fusions. PMID- 15549484 TI - Implant removal for late-developing infection after instrumented posterior spinal fusion for scoliosis: reinstrumentation reduces loss of correction. A retrospective analysis of 45 cases. AB - A retrospective follow-up study of patients who, having undergone instrumented posterior spinal fusion for scoliosis, experienced late infection and then underwent either implant removal alone or implant removal and instrumented refusion. We conducted this study to determine whether it is possible to avoid loss of correction by a single-stage implant removal and reinstrumentation procedure. There have been a few reports of late-appearing infections after spinal instrumentation. Implant bulk, metallurgic reactions, and contamination with low-virulence microorganisms have been suggested as possible etiologic factors. The clinical symptoms include pain, swelling, redness, and spontaneous drainage of fluid. Complete instrumentation removal and systemic antibiotics is usually curative. We retrospectively reviewed 45 patients who underwent instrumented posterior spinal fusion for scoliosis and experienced development of late infections and, after a mean of 3 years after the initial procedure, either underwent implant removal alone [ n=35, instrumentation removal (HR) group] or additionally underwent reinstrumentation and fusion [ n=10, reinstrumentation and fusion (RI&F) group]. Three patients were reinstrumented 1.5 years after instrumentation removal, and seven underwent a one-stage rod removal and reinstrumentation/refusion procedure. Allergic predisposition, protracted postoperative fever, and pseudarthrosis appear to increase the risk of late developing infection after posterior spinal fusion. All wounds in both groups healed uneventfully. Preoperative radiographic Cobb measurements showed no statistically significant between-group differences. At follow-up, however, outcome was clearly better in the RI&F group: Loss of correction was significantly smaller in reinstrumented patients. Thus, the thoracic Cobb angle was 28+/-16 degrees (range 0-55 degrees ) in the RI&F group versus 42+/-15 degrees (21-80 degrees ) in the HR group, and the lumbar Cobb angle was 22+/-11 degrees (10-36 degrees ) in the RI&F group versus 29+/-12 degrees (13-54 degrees ) in the HR group. The results of our study demonstrate that wound healing is usually uneventful after instrumentation removal for late infection, also when patients undergo instrumented refusion in a one-stage procedure. Reinstrumentation appears to achieve permanent correction of scoliosis. PMID- 15549486 TI - TNFalpha-stimulated gene product (TSG-6) and its binding protein, IalphaI, in the human intervertebral disc: new molecules for the disc. AB - Inflammation and irritation of the nerve roots has been indicated as an important factor in the pain associated with symptomatic disc herniations. Tumour necrosis factor alpha (TNFalpha) is now believed to be involved in this pathway. TNFalpha causes connective tissue cells in culture to synthesise a glycoprotein, TNFalpha stimulated gene-6 (TSG-6). TSG-6 is found in inflammatory diseases of related connective tissues, such as articular cartilage in rheumatoid arthritis, but is not present in unaffected individuals. In order to determine if TSG-6 occurred in intervertebral disc (and cartilage endplate), we have investigated the presence of TSG-6 and its binding protein, inter-alpha-inhibitor (IalphaI), in 58 herniated and 15 non-herniated discs. Immunostaining for the cytokines, IL 1alpha, IL-1beta and TNFalpha, has also been carried out. We have demonstrated that both TSG-6 and IalphaI occur commonly in human intervertebral disc matrix with at least some TSG-6 in 98% of discs studied and IalphaI in all of them. Staining for TSG-6 was greatest in herniated discs, particularly close to blood vessels. IalphaI immunostaining was frequently widespread throughout the disc but there was little in the cartilage endplate. It has been proposed that these molecules have widespread effects, including extracellular matrix stabilisation, down-regulation of the protease network and reduction of inflammation. Hence, the occurrence of TSG-6 and IalphaI in disc tissue could have implications in the aetiopathogenesis and future therapeutics of intervertebral disc disease. PMID- 15549487 TI - The value of electromyography of the lumbar paraspinal muscles in discriminating between chronic-low-back-pain sufferers and normal subjects. AB - The authors studied the surface electromyographic (EMG) spectrum of the paraspinal muscles of 350 subjects. They were classified by their history as normal (n=175), chronic low back pain (n=145), or past history (n=30). They pulled upwards on a floor-mounted load cell at two-thirds of their maximum voluntary contraction for 30 s, while the EMG was measured from the paraspinal muscles at the L4/L5 level. From the EMG signal the root-mean-square (RMS) was calculated. Power spectrum analysis allowed calculation of the median frequency slope, the initial median frequency (IMF), modal frequency, peak amplitude and spectral width at half peak amplitude (half-width). All of the variables of the chronic group were significantly different from the normal group, except the median frequency slope, RMS slope and mode. Half-width, age and maximum voluntary contraction were shown to be independent predictors of back pain classification. Half-width classified the subjects with a sensitivity of 0.65 and a specificity of 0.75. PMID- 15549489 TI - Sequence variation in the gene encoding the major capsid protein of Korean fish iridoviruses. AB - Ten iridoviruses were isolated from cultured fish from various regions in Korea; 7 from rock bream, 1 from red sea bream, 1 from sea bass, and 1 from rockfish. The full open-reading frame (ORF) encoding the major capsid protein (MCP) (1362 bp) from ten iridoviruses were sequenced and the nucleotide sequences were phylogenetically analyzed. Phylogenetic analysis revealed that the ten Korean isolates were classified into one cluster. However, their sequences were not identical and, based on the nucleotide sequence variation, they could be further divided into two subgroups. While nine Korean isolates were similar to the Japanese isolate red sea bream iridovirus (RSIV), one isolate was distinct from other iridovirus isolates. These results suggest that a diversity of iridoviruses exist in Korea and that a new variant strain has emerged. PMID- 15549490 TI - Differential protein expression profile in gastrointestinal stromal tumors. AB - Gastrointestinal stromal tumors (GISTs) arise from the interstitial cells of Cajal through gain of function mutations of the oncogene KIT. Imatinib offers the first effective treatment for patients with GISTs, but the therapeutic outcome strongly depends on the type of KIT mutation. We used ProteinChip technology to investigate whether GISTs with different KIT mutations express different proteins. In total, 154 proteins were significantly differentially expressed in GISTs with exon 9 KIT mutation compared to GISTs with exon 11 KIT mutation. PMID- 15549491 TI - Characteristics of basal taurine release in the rat striatum measured by microdialysis. AB - Taurine is a sulfur-containing amino acid thought to be an osmoregulator, neurotransmitter or neuromodulator in the brain. Our objective was to establish how much taurine is released in the striatum and examine the mechanisms controlling extracellular taurine concentrations under resting conditions. The experiments were made on rats by microdialysis in vivo. Changes in taurine were compared with those in glutamate, glycine and the non-neuroactive amino acid threonine. Using the zero net flux approach we showed the extracellular concentration of taurine to be 25.2 +/- 5.1 muM. Glutamate was increased by tetrodotoxin and decreased by Ca2+ omission, glycine and threonine were not affected and both treatments increased extracellular taurine. The basal taurine release was increased by the taurine transport inhibitor guanidinoethanesulfonate and reduced by the anion channel blocker 4-acetamido-4'-isothiocyanatostilbene 2,2'-disulfonic acid. PMID- 15549492 TI - The HPLC resolution of N-2,4-dinitrophenylated amino acids and peptides stereoisomers on naphthylethylcarbamate-beta-cyclodextrin bonded phase using the acetonitrile-based mobile phase: evidence for the chiral recognition pattern. AB - A facile method of enantioresolving a variety of alpha-amino acids and peptides on naphthylethylcarbamate-beta-cyclodextrin bonded phases (i.e., SN- and RN-beta CDs) under the elution of acetonitrile-based mobile phase makes use of 2,4 dinitrofluorobenzene (DNFB) as the tagging reagent, which undergoes nucleophilic substitution by the free amino group in alkaline medium to give a N-2,4 dinitrophenyl (DNP) derivative. The resolution is better obtained on RN-beta-CD phase and fails to reproduce on the intact beta-cyclodextrin bonded phase under the same chromatographic conditions, which strongly suggests that the observed resolution should be due to the interaction of analyte with naphthylethylcarbamate moiety, not with the residual secondary hydroxyl groups on the beta-cyclodextrin. PMID- 15549493 TI - Hexafluoroacetone as protecting and activating reagent: site-selective functionalization of iminodiacetic acid. AB - Hexafluoroacetone was applied as a bidentate protecting and activating agent for the syntheses of RGD-peptide mimetics starting from iminodiacetic acid in solution and on solid phase. PMID- 15549494 TI - Structural and biochemical implications of single amino acid substitutions in the nucleotide-dependent switch regions of the nitrogenase Fe protein from Azotobacter vinelandii. AB - The structures of nitrogenase Fe proteins with defined amino acid substitutions in the previously implicated nucleotide-dependent signal transduction pathways termed switch I and switch II have been determined by X-ray diffraction methods. In the Fe protein of nitrogenase the nucleotide-dependent switch regions are responsible for communication between the sites responsible for nucleotide binding and hydrolysis and the [4Fe-4S] cluster of the Fe protein and the docking interface that interacts with the MoFe protein upon macromolecular complex formation. In this study the structural characterization of the Azotobacter vinelandii nitrogenase Fe protein with Asp at position 39 substituted by Asn in MgADP-bound and nucleotide-free states provides an explanation for the experimental observation that the altered Fe proteins form a trapped complex subsequent to a single electron transfer event. The structures reveal that the substitution allows the formation of a hydrogen bond between the switch I Asn39 and the switch II Asp125. In the structure of the native enzyme the analogous interaction between the side chains of Asp39 and Asp125 is precluded due to electrostatic repulsion. These results suggest that the electrostatic repulsion between Asp39 and Asp125 is important for dissociation of the Fe protein:MoFe protein complex during catalysis. In a separate study, the structural characterization of the Fe protein with Asp129 substituted by Glu provides the structural basis for the observation that the Glu129-substituted variant in the absence of bound nucleotides has biochemical properties in common with the native Fe protein with bound MgADP. Interactions of the longer Glu side chain with the phosphate binding loop (P-loop) results in a similar conformation of the switch II region as the conformation that results from the binding of the phosphate of ADP to the P-loop. PMID- 15549496 TI - One-year clinical evaluation of a flowable resin liner associated with a microhybrid resin in noncarious cervical lesions. AB - The aim of this study was to compare the clinical performance over 1 year of a microhybrid composite resin for class V restorations both lined and not lined with a flowable composite resin. Nineteen patients having at least two pairs of cervical noncarious lesions under occlusion were enrolled in this study. A total of 38 restorations were placed, half for each group (Single-Bond + Filtek-Flow + Filtek Z250, and Single-Bond + Filtek Z250). Two calibrated operators placed all restorations according to the manufacturers' instructions. Two other independent examiners evaluated the restorations at baseline and after a 12-month period according to the USPHS criteria and modified criteria for color match. The classic alpha score was divided into A1 for "not detectable" and A2 for "slightly discernible" filling. Statistical analysis was conducted using Fisher's exact test and McNemar's test (P=0.05). One restoration was lost after 12 months for each group (retention rate 95% for each group). After 12 months, 18 restorations showed a trend towards dark yellowing (color match A2). The use of Filtek Flow as a liner under Filtek Z250 restorations did not improve the clinical performance of class V restorations after 6 and 12 months of evaluation. PMID- 15549497 TI - Approach to a giant inguinoscrotal hernia. AB - We present an extremely huge and longstanding, giant inguinoscrotal hernia extending to below the knee with an ulcer at its base. Though hernias of this magnitude are rare, their management can be demanding and challenging. Nevertheless, in an emergency situation, the repair of the hernial defect is not essential, especially in a compromised patient. In fact, the most important step is excision of the devitalised tissue, and the final surgery can be delayed. In nonemergency management, definitive surgery can be planned either by a period of preoperative staged pneumoperitoneum, repairing after a resection of bowel and omentum, or replacing the content and ventilation of the patient to avoid the pressure on the cardiorespiratory system by forcing the tissue. This case highlights the problems encountered in management of huge hernias. PMID- 15549498 TI - Operative treatment of massive ventral hernia using polypropylene mesh: a challenge for surgeon and anesthesiologist. AB - OBJECTS: Surgical repair of very large ventral hernias has become feasible after the introduction of synthetic meshes and developments in intensive-care treatment. In addition to the operative challenges, postoperative disorders in the cardiovascular system, tissue oxygenation, increased intra-abdominal pressure, and pulmonary embolism expose the patient to severe risks. METHODS: From 1997-2002 we operated on ten patients with giant ventral incisional or umbilical hernia (mean defect size 240 cm(2)) by using retromuscular polypropylene mesh. All patients were morbidly obese [mean Body Mass Index (BMI) 39+/-7.2 kg/m(2)]. Four of the operations were emergencies because of an acute intestinal occlusion, bowel gangrene, and skin complications. The patients were reinvestigated after the mean follow-up of 2.5 years to find out the frequency of recurrence and degree of disability. RESULTS AND CONCLUSION: There was no intraoperative mortality, but one patient died at home after 5 weeks because of myocardial infarct and prolonged wound infection. She had mild stable coronary heart disease preoperatively. Although minor wound complications were observed in three patients, there was no need to remove the meshes. One small recurrent hernia was observed in the follow-up, but it was too small to be repaired. The quality of life after surgery was good for all patients, and they were satisfied with the operation. Retromuscular mesh hernioplasty associated with careful patient monitoring in intensive care is safe and feasible in the selected patients with massive ventral hernia. PMID- 15549499 TI - Population prevalence of APOE, APOC3 and PPAR-alpha mutations associated to hypertriglyceridemia in French Canadians. AB - Hypertriglyceridemia (HTG) is known as a common metabolic disorder associated with increased production, decrease catabolism and/or decreased hepatic uptake of triglyceride (TG)-rich particles. We assessed, in the Quebec City population, the allele frequency and haplotype distributions of mutations in genes related to HTG, such as the apolipoprotein E (APOE) (C112R and C158R), the apolipoprotein CIII (APOC3) (C-482T and C3238G) and the peroxisome proliferator-activated receptor alpha (PPARalpha) (L162V) genes. A total of 938 anonymous unlinked newborns from the metropolitan Quebec City area have been genotyped. Allele frequencies observed in the Quebec City population differed from known frequencies determined in other Caucasian populations. The co-transmitted allele distribution between the two-marker genotypes APOE/APOC3(C3238G) and APOC3(C 482T)/PPARalpha(L162V) presented a weak deviation from the assumption of genetic independence. Also, we observed a non-independent distribution of the T-482/G3238 allele combinations within the APOC3 gene, suggesting strong linkage disequilibrium between the C-482T and C3238G polymorphisms. Moreover, comparisons of allele frequencies observed in the population of Quebec City to those obtained in other Caucasian populations suggested that the population of Quebec City may be at a lower risk of developing HTG due to APOE, APOC3 and PPARalpha genetic variants. However, the strong linkage disequilibrium and the two-marker genotype distributions observed in the APOC3 gene suggest that these two variants may functionally interact in the Quebec City population. PMID- 15549500 TI - Association of an intragenic microsatellite marker in the CC16 gene with asthma in the Indian population. AB - The gene for Clara cell secretory protein (CC16) is an ideal candidate for investigating genetic predisposition to asthma because of its role in the airway as an anti-inflammatory molecule, differences in its levels between asthmatics and healthy controls, and its genetic location (11q13). We investigated the association of an SNP (A38G) and an intragenic repeat polymorphism in the CC16 gene with asthma and its associated traits, such as total serum IgE levels, in a case control as well as in a family based study design. A significant association was observed for the microsatellite repeat at the level of alleles and genotypes with asthma (P<0.05) in both the study designs. However, no association was observed for the A38G SNP with asthma. When haplotypes were constructed for these two loci and compared, the haplotype A_18 was found at higher frequency in patients (OR=1.59, 95%CI=1.08, 2.33, P=0.016). Also, in the family based design, a biased transmission was observed for haplotypes from parents to affected offspring (P=0.003). Individually, haplotype A_18 showed preferential transmission (82.6%) to affected offspring (P=0.001), thereby confirming the case control results. In summary, this is the first study identifying the CC16 gene to be associated with asthma in the Indian population. PMID- 15549502 TI - Causal knowledge and imitation/emulation switching in chimpanzees (Pan troglodytes) and children (Homo sapiens). AB - This study explored whether the tendency of chimpanzees and children to use emulation or imitation to solve a tool-using task was a response to the availability of causal information. Young wild-born chimpanzees from an African sanctuary and 3- to 4-year-old children observed a human demonstrator use a tool to retrieve a reward from a puzzle-box. The demonstration involved both causally relevant and irrelevant actions, and the box was presented in each of two conditions: opaque and clear. In the opaque condition, causal information about the effect of the tool inside the box was not available, and hence it was impossible to differentiate between the relevant and irrelevant parts of the demonstration. However, in the clear condition causal information was available, and subjects could potentially determine which actions were necessary. When chimpanzees were presented with the opaque box, they reproduced both the relevant and irrelevant actions, thus imitating the overall structure of the task. When the box was presented in the clear condition they instead ignored the irrelevant actions in favour of a more efficient, emulative technique. These results suggest that emulation is the favoured strategy of chimpanzees when sufficient causal information is available. However, if such information is not available, chimpanzees are prone to employ a more comprehensive copy of an observed action. In contrast to the chimpanzees, children employed imitation to solve the task in both conditions, at the expense of efficiency. We suggest that the difference in performance of chimpanzees and children may be due to a greater susceptibility of children to cultural conventions, perhaps combined with a differential focus on the results, actions and goals of the demonstrator. PMID- 15549503 TI - Normalisation of serum cholestanol concentration in a patient with cerebrotendinous xanthomatosis by combined treatment with chenodeoxycholic acid, simvastatin and LDL apheresis. AB - The concentrations of serum cholesterol, cholestanol and non-cholesterol sterols were measured in a patient with cerebrotendinous xanthomatosis under different therapeutic regimens. During treatment with chenodeoxycholic acid (CDCA) (750 mg/day) plus simvastatin (20 mg/day) for two years cholesterol and cholestanol concentrations averaged 188+/-10 mg/dl and 0.54+/-0.03 mg/dl. Thereafter treatment with simvastatin was discontinued. During treatment with low-density lipoprotein (LDL)-apheresis plus CDCA for 33 weeks, cholestanol concentrations reached almost normal levels (0.48+/-0.03 mg/dl immediately before and 0.32+/ 0.02 mg/dl directly after LDL-apheresis, n=6). A further reduction of cholesterol and cholestanol was achieved by addition of simvastatin (20 mg/day). Cholesterol and cholestanol concentrations before and after LDL-apheresis during this treatment period averaged 122+/-4 mg/dl and 55+/-10 mg/dl, and 0.42+/-0.02 mg/dl and 0.18+/-0.06 mg/dl, respectively. Despite the consistent reduction of cholestanol to normal or even subnormal levels, a definite improvement of clinical symptoms was not noted. Our results suggest caution in the recourse to an aggressive cholestanol lowering therapy. PMID- 15549501 TI - Development and validation of the osteoporosis prescreening risk assessment (OPERA) tool to facilitate identification of women likely to have low bone density. AB - Osteoporosis and its consequent increase in fracture risk is a major health concern for postmenopausal women and older men and has the potential to reach epidemic proportions. The "gold standard" for osteoporosis diagnosis is bone densitometry. However, economic issues or availability of the technology may prevent the possibility of mass screening. The goal of this study was to develop and validate a clinical scoring index designed as a prescreening tool to help clinicians identify which women are at increased risk of osteoporosis [bone mineral density (BMD) T-score -2.5 or less] and should therefore undergo further testing with bone densitometry. Records were analyzed for 1522 postmenopausal females over 50 years of age who had undergone testing with dual-energy X-ray absorptiometry (DXA). Osteoporosis risk index scores were compared to bone density T-scores. Hologic QDR 4500 technology was used to measure BMD at the femoral neck and lumbar spine (L1-L4). Participants who had a previous diagnosis of osteoporosis or were taking bone-active medication were excluded. Receiver operating characteristic (ROC) analysis was used to identify the specific cutpoint value that would identify women at increased risk of low BMD. A simple algorithm based on age, weight, history of previous low impact fracture, early menopause, and corticosteroid therapy was developed. Validation of this five-item osteoporosis prescreening risk assessment (OPERA) index showed that the tool, at the recommended threshold (or cutoff value) of two, had a sensitivity that ranged from 88.1 [95% confidence interval (CI) for the mean: 86.2-91.9%] at the femoral neck to 90% (95% CI for the mean: 86.1-93.1%) at the lumbar spine area. Corresponding specificity values were 60.6 (95% CI for the mean: 57.9-63.3%) and 64.2% (95% CI for the mean: 61.4-66.9%), respectively. The positive predictive value (PPV) ranged from 29 at the femoral neck to 39.2% at the lumbar spine, while the corresponding negative predictive values (NPVs) reached 96.5 and 96.2%, respectively. Based on this cutoff value, the area under the ROC curve was 0.866 (95% CI for the mean: 0.847-0.882) for the lumbar spine and 0.814 (95% CI for the mean: 0.793-0.833) for the femoral neck. We conclude that the OPERA is a free and effective method for identifying Italian postmenopausal women at increased risk of osteoporosis. Its use could facilitate the appropriate and more cost-effective use of bone densitometry in developing countries. PMID- 15549504 TI - The role of brain infarcts and hippocampal atrophy in subcortical ischaemic vascular dementia. AB - We investigated if, in patients with vascular lesions, the variable that best discriminated demented from non-demented patients was the severity of the vascular pathology or the degree of hippocampal atrophy. A total of 39 patients multiple subcortical infarcts, who could be considered as possible vascular dementia with small vessel pathology, with underwent a neuropsychological study and brain magnetic resonance imaging (MRI) DSM IV criteria supported by neuropsychological data were used to distinguish demented from non-demented patients. The MRI study took into account the degree of hippocampal atrophy (hippocampal height and interuncal distance) and the severity of vascular pathology (number of brain infarcts). The distribution of lesions and a factor analysis showed that hippocampal atrophy is a better predictor of dementia than the number of brain infarcts. Multiple subcortical infarcts alone are probably not able to cause clinical dementia but the presence of vascular lesions increases the expression of concomitant Alzheimer's disease. PMID- 15549505 TI - Cognitive deterioration in Alzheimer's disease: is the early course predictive of the later stages? AB - This study investigated the predictive value of the early progression rate of Alzheimer's disease on that of the later stages. We retrospectively evaluated 91 patients affected by possible Alzheimer's disease; the patients had been examined twice with the Milan overall dementia assessment (MODA) scale at an interval of 12 months (53 patients) or 24 months (38 patients). At the first assessment, speed of progression was calculated for each patient dividing the MODA difference from the normality threshold by the time elapsed from the cognitive decline onset. The second assessment of speed of progression was calculated dividing the difference between the two MODA scores by the time elapsed from the first examination. Patients with a slow progression rate in the early stage were unlikely to show a subsequent fast progression rate, and vice versa for patients with a fast early progression. A tool is provided for predicting the speed of cognitive decline of patients from a single MODA assessment. We suggest that, in future clinical trials on Alzheimer's disease, a stratification of participants based on the early rate of cognitive deterioration may be considered. PMID- 15549506 TI - "What do you remember about Chernobyl?" A new test of memory for media-mediated events. AB - We devised a new test for assessing remote memory for media-mediated events, i.e. events that are reported by and known through the media. The test consists of a verbal questionnaire covering famous events that occurred over a long period of time, from 1976 to 2000, specifically designed for use with the Italian population. A free recall procedure was adopted which makes use of progressive cues. The test can be easily updated for inclusion of future events. Normative and test-retest reliability data are presented. The test provides a new tool for assessing media-mediated memory whenever an extensive assessment of retrograde memory performance is required, both in normal people and in brain-damaged patients. PMID- 15549507 TI - A case of isolated tuberculoid leprosy of antebrachial medial cutaneous nerve. AB - Leprosy is an infectious disease of prevalence still high in endemic areas in Brazil. The neurological presentation depends on the involved nerve and is usually associated with skin lesions and the formation of multiple abscesses. We present a case of isolated tuberculoid leprosy, discuss the occurrence, the differential diagnosis and the treatment of this rare presentation and reaffirm the importance of considering leprosy in the differential diagnosis of patients with polyneuropathy or nerve enlargement with no skin lesions. PMID- 15549508 TI - Pure post-stroke cerebellar cognitive affective syndrome: a case report. AB - Cerebellar pathology commonly shows important motor signs and less evident cognitive dysfunction. The 'cerebellar cognitive affective syndrome' is characterised by impairment on executive function, spatial cognition, language and behaviour. We report the case of a man with acute onset of transitory motor features and severe mental disorders. Cranial CT and brain MRI revealed extended cerebellar lesions. Neuropsychological assessment disclosed deficits of attention, executive function and memory. Auditory event-related potentials showed abnormal P300. These data suggest a pure "cerebellar cognitive affective syndrome"and strengthen the hypothesis of cerebellar cognitive function modulation. PMID- 15549509 TI - Subcortical nodular heterotopia: a functional MRI and somatosensory evoked potentials study. AB - Subcortical nodular heterotopia (SNH) associated with refractory epilepsy may be surgically treated, and a positive outcome can be expected following the complete excision of the malformed tissue. Recent functional neuroimaging studies have suggested the possible functional relevance of cerebral malformations, and may make it possible to improve presurgical planning, thus allowing extended resections and minimising post-operative deficits. We here report the case of a 19-year-old man with epilepsy and a giant SNH associated with diffused abnormal gyrations of the right temporal-parietal regions. Cortical functional organisation was investigated by means of functional magnetic resonance imaging (MRI) during sensory and motor tasks, and somatosensory evoked potentials. The results revealed enlarged and displaced motor and sensory cortical areas with heterotopic tissue functional activation. The relevance of these findings is discussed in the light of the possible surgical treatment of drug-refractory epilepsy associated with cerebral malformations: surgical treatment based on conventional MRI studies alone, without taking the functional nature of dysplastic tissues into account, may lead to considerable side effects. PMID- 15549510 TI - Ischaemic stroke in progressive systemic sclerosis. AB - Progressive systemic sclerosis (PSS) or scleroderma is a multisystem disease affecting the skin, lungs, myocardium, kidneys and gastrointestinal tract. Primary involvement of cerebral arteries in PSS has been reported but is very rare. A 61-year-old woman suffering from scleroderma for six years was hospitalised for two subsequent episodes of transient acute dysarthria and left hemiparesis. After five hours from the first onset of symptoms, she was submitted to brain magnetic resonance (MR) protocol that showed a right subinsular ischaemic lesion and whole right middle cerebral artery (MCA) territory hypoperfusion. Intracranial and epiaortic MR angiography reported a focal stenosis in the M2 portion of MCA. She was immediately treated with i.v. high dose steroids and oral acetylsalicylic acid. At one-month follow up, MR findings were confirmed. We have documented a cerebral infarct in a PSS patient. In our opinion, the ischaemic stroke was caused by a localised autoimmune angiopathy. PMID- 15549511 TI - Sporadic Creutzfeldt-Jakob disease without dementia at onset: clinical features, laboratory tests and sequential diffusion MRI (in an autopsy-proven case). AB - A rapidly progressing dementia, followed by focal neurological signs, and evidence of periodic sharp wave complexes (PSWC) in the EEG may lead to the clinical suspicion of Creutzfeldt-Jakob disease (CJD). Different clinical variants of CJD have been described in the past, with prominent extrapyramidal or occipital lobe involvement, all included in the sporadic form of CJD (sCJD). Familiar and iatrogenic forms of CJD are also known. More recently a new variant has been described, vCJD, casually linked to bovine spongiform encephalopathy (BSE) and it has attracted increasing attention toward each form of rapidly progressing dementia; likewise the differential diagnosis between sCJD vs. vCJD is not always easy. Magnetic resonance imaging (MRI) too seems to have a peculiar role in differentiating sCJD from vCJD, even if the role of MRI in the diagnosis of CJD is still debated. Diffusion MRI is expected to play an important role in the clinical setting of CJD, contributing to formulation of an early diagnosis, especially in cases with unusual clinical presentation. In fact, the sensitivity of diffusion MRI is superior to that of conventional MRI (T1, T2, FLAIR) in detecting specific basal ganglia and cortical abnormalities early in the course of CJD and these abnormalities correlate well with areas of the most severe and characteristic neuropathological changes. We describe a case of autopsy-proven sCJD, with an unusual clinical course without dementia as a presenting symptom and discuss the role of diffusion MRI and laboratory tests in making an early diagnosis. PMID- 15549512 TI - Skeletal muscle involvement in congenital long QT syndrome. AB - Skeletal muscle involvement in a genetically confirmed congenital long QT syndrome (LQTS) has not been reported. In a 36-year-old male, intracerebral bleeding from a brainstem cavernoma suddenly occurred. His history was noteworthy for congenital LQTS due to a point mutation in the KVLQT1-gene on chromosome 11. The mutation was also found in his two daughters, his sister, her daughter and his mother. His ECG showed sinusrhythm and a QTc-interval of 0.46 ms. Needle-EMG from the right abductor pollicis, left brachial biceps, left extensor digitorum, and right anterior tibial muscle(s), carried out because muscle involvement had been previously reported in clinically diagnosed congenital LQTS, revealed abnormal spontaneous activity, but otherwise normal motor unit architecture. LQTS, due to a KVLQT1 mutation, may manifest not only in the heart, but subclinically also in the skeletal muscle. PMID- 15549513 TI - Superselective angiography increases the diagnostic yield in the investigation of intracranial haematomas caused by micro-arteriovenous malformations. PMID- 15549514 TI - Disability and quality of life in different primary headaches: results from Italian studies. AB - Headaches may have a wide range of impact on patients' lives. We report the results of Italian studies in which disability and health-related quality of life (HRQOL) in patients with different primary headaches were evaluated. The Short Form 36 (SF-36) was used to assess HRQOL; the Migraine Disability Assessment Score questionnaire (MIDAS) was used to assess disability in patients with migraine without aura or with chronic migraine. Mean MIDAS total scores were evaluated in migraine without aura and chronic migraine patients. The scores at the eight SF-36 scales were calculated in patients with the three studied headaches, and were compared with Italian normative data (Student's t-test with Bonferroni correction). Primary headaches had a considerable negative impact on patients' lives, with poor quality of life and decreased ability to function in daily duties. The mean MIDAS total score was 23.4 in 264 patients with migraine without aura, and 79.2 in 150 patients with chronic migraine. Mean SF-36 scores in migraine without aura (68 subjects), chronic migraine (84) and cluster headache (56) were lower than those from the Italian general population, with significant differences for 3 scales in migraine without aura, for 6 in chronic migraine, and for all scales in cluster headache. Our results confirmed a marked personal and social burden in patients with migraine without aura, and also in the less well-studied forms of primary headaches, cluster headache and chronic migraine. PMID- 15549515 TI - Measuring responses to therapy in headache patients: new and traditional end points. AB - Changes in numerical indices of the intensity, duration and frequency of attacks are traditionally used to assess the effectiveness of treatments for migraine and other primary headaches. However, recent clinical and therapeutic guidelines and guidelines for conducting clinical trials specify that the essential aim of symptomatic and prophylactic migraine treatments should be to reduce the global impact of the headaches on the patient's life. Some standardised instruments for assessing disability and health-related quality of life seem sensitive to treatment-induced changes in primary headache patients. However, further studies to determine the suitability of these instruments as outcome measures in clinical practice are necessary. PMID- 15549516 TI - Disability in young patients suffering from primary headaches. AB - During the last decade researchers have begun to employ standardised methodologies to investigate the global impact of primary headaches. Disease specific instruments have been developed to measure headache-related disability. The MIDAS Questionnaire, which is the most extensively studied of these instruments, was designed to assess the overall impact of headaches over the 3 months before compilation. The MIDAS Questionnaire is an optimal tool to assess headache-related disability in adults in relation to patients' daily activities. Primary headaches are a recurrent problem for children and adolescents. Forty percent of children have experienced headaches by the age of 7 years increasing to 75% by the age of 15. In a recent report we determined the suitability of the MIDAS Questionnaire in its original form for assessing disability in children and adolescents suffering from different kinds of headache. This was the first step of a line of research aimed to develop a new MIDAS Questionnaire adapted for young patients. In this second study the aims were: (1) to produce a new version of the MIDAS Questionnaire specific for young patients suffering from different forms of headache; (2) to assess the reliability of this new instrument; (3) to assess its sensitivity to treatment intervention. PMID- 15549517 TI - Acute drug treatment of migraine attack. AB - The availability of an effective drug for attack treatment is a main need of migraine patients. Symptomatic therapy of migraine now includes three main classes of drugs: ergot alkaloids, nonsteroidal antiinflammatory drugs (NSAIDs) and triptans. Clinical randomised placebo-controlled trials affirmed the efficacy and tolerability of these drugs, when used at the recommended doses and in the absence of contraindications. The efficacy data from comparative trials showed a trend in favour of triptans vs. ergot alkaloids, but failed to show significant differences between triptans and NSAIDs. However, clinical practice experience of most headache clinicians suggests that triptans provide superior efficacy in comparison with nonspecific agents, mainly in that they better satisfy migraine patient expectations. In fact, these selective specific antimigraine drugs are an advance in acute migraine treatment. PMID- 15549518 TI - Cluster headache: symptomatic treatment. AB - The clinical management of cluster headache (CH) attacks requires a symptomatic treatment that is rapidly effective in resolving or significantly reducing symptoms. First-choice drugs for the symptomatic treatment of CH are subcutaneous sumatriptan at a dose of 6 mg and 100% oxygen inhalation at a rate of 7 l/min for no more than 15 min. Sumatriptan acts by suppressing pain and the accompanying autonomic phenomena, with no substantial differences in its mechanism of action between episodic and chronic CH. The drug can be used for prolonged periods without loss of efficacy or safety and its side-effects are generally mild or moderate. Oxygen inhalation has a number of advantages over drug therapy: it is free from side-effects, has no contraindications--unlike sumatriptan, it can be used in patients with cardiac, cerebral or peripheral vascular disease and with kidney, liver or lung disease--acts rapidly and can be administered several times a day. Its disadvantages are that it is scarcely practical and may induce a "rebound effect". Sumatriptan nasal spray, zolmitriptan and dihydroergotamine nasal spray are scarcely effective. After the introduction of sumatriptan, ergotamine tartrate has been relegated to a secondary role in the symptomatic treatment of CH. Among other non-drug and topical drug treatment options, hyperbaric oxygen therapy and the intranasal application of 10% cocaine hydrochloride and 10% lidocaine in the sphenopalatine fossa have also proved effective. PMID- 15549519 TI - Migraine and stroke. AB - The association between migraine and stroke is a dilemma for neurologists. Migraine is associated with an increased stroke risk and it is considered an independent risk factor for ischaemic stroke in a particular subgroup of patients. The pathogenesis is not known but several studies report some common biochemical mechanisms between the two diseases. A classification of migraine related stroke that encompasses the full spectrum of the possible relationship between migraine and stroke has been proposed and it includes three main entities: coexisting stroke and migraine, stroke with clinical features of migraine, and migraine-induced stroke. The concept of migraine-induced stroke is well represented by migrainous infarction; it is described in the revised classification of the International Headache Society (IHS), and it represents the strongest demonstration of the relationship between ischaemic stroke and migraine. A very interesting common condition in stroke and migraine is patent foramen ovale (PFO) which could play a pathogenetic role in both disorders. The association between migraine and cervical artery dissection (CAD) is reported in recent studies. Migraine is more frequent in patients with CAD. This supports the hypothesis that an underlying arterial wall disease could be a predisposing condition for migraine. The neuroradiological evidence of subclinical lesions most typical in the white matter and in the posterior artery territories in patients with migraine, opens a new field of research. In conclusion the association between migraine and stroke remains conflicting. Solving the above mentioned issues is fundamental to understanding the epidemiologic, pathogenetic and clinical aspects of migraine-related stroke. PMID- 15549520 TI - Coagulation abnormalities in migraine and ischaemic cerebrovascular disease: a link between migraine and ischaemic stroke? AB - Migraine, particularly migraine with aura, is a risk factor for ischaemic stroke. The mechanisms underlying this association are obscure. One hypothesis is that shared risk factors may be the cause of this association. Over the last decade, studies have suggested an association between migraine and genetic abnormalities in coagulation factors which play an important role in stroke pathogenesis. Although the results of studies on various prothrombotic conditions are conflicting, findings suggest a higher frequency of some genetic abnormalities in migraine with aura patients. Thus, persistent hypercoagulability may explain the tendency for these patients to develop thromboembolic cerebrovascular events, especially when they are exposed to additional procoagulant stresses. Further studies on larger samples are required to test this hypothesis. PMID- 15549521 TI - The role of cardiac diseases in the comorbidity between migraine and stroke. AB - Several case-control and cohort studies have suggested an association between migraine and stroke. A significantly higher risk for stroke was found in women under the age of 45 years and for the subgroup with migraine with aura, the posterior circulation being significantly more frequently involved. The link between cardiac diseases and the comorbidity migraine-stroke has been evaluated considering both possible relationships: a higher prevalence of a vascular disease involving both heart and brain in migraineurs, or a cardiac disorder, more prevalent in migraineurs, with a possible aetiological role in migraine attacks. PMID- 15549522 TI - Headache and hypertension. AB - Headache is generally regarded as a symptom of high blood pressure in spite of conflicting opinions on the association of headache and arterial hypertension. Most studies have shown that mild chronic hypertension and headache are not associated and this demonstration needs to be implemented in clinical practice. Whether moderate hypertension predisposes to headache remains controversial, but there is little evidence that it does. Headaches caused by significant disturbances in arterial pressure are included in the section "Headache attributed to a disorders of homeostasis" (code 10.3) of the 2nd edition of the International Classification of Headache disorders. PMID- 15549523 TI - Spontaneous low cerebrospinal pressure: a mini review. AB - Spontaneous intracranial hypotension (SIH) is a syndrome of low cerebrospinal fluid (CSF) pressure characterised by postural headaches in patients without any history of dural puncture or penetrating trauma. Described by Schaltenbrand in 1938, SIH is thought to result from an occult CSF leak resulting in decreased CSF volume and, consequently, in low CSF pressure. Magnetic resonance imaging of the head and spine has improved the diagnosis of the syndrome showing peculiar radiographic abnormalities including diffuse pachymeningeal enhancement, subdural fluid collections and downward displacement of the cerebral structures. Treatment of SIH headache should start with conservative, non-invasive therapies while epidural blood patch has emerged as the treatment of choice for those symptomatic patients who have failed medical noninvasive approaches. PMID- 15549524 TI - Headache and intracranial hypotension: neuroradiological findings. AB - The cardinal and classic features of postural headache and low cerebrospinal fluid (CSF) pressure in intracranial hypotension may not dominate the clinical picture of the syndrome and may be associated with additional various neurological symptoms and signs. Reports of unusual clinical presentations continue to appear in the literature. Despite the considerable variability of the clinical spectrum, neuroradiological studies reveal more constant and characteristic features. Brain MRI findings include intracranial pachymeningeal thickening and post-contrast enhancement, subdural fluid collections and downward displacement or "sagging" of the brain. Spinal MRI findings include collapse of the dural sac with a festooned appearance, intense epidural enhancement owing to dilatation of the epidural venous plexus, and possible epidural fluid collections. In fact, spinal studies may demonstrate CSF leakage from spinal dural defects, which are considered the most common cause of the syndrome. Myelo MR may suggest the possible point of CSF leakage, by demonstrating an irregular root sleeve; myelo-CT and radioisotope myelocisternography (RMC) are often needed to confirm the point of CSF leakage. Neuroimaging studies are, therefore, essential for suggesting and confirming the diagnosis. PMID- 15549525 TI - Headache in brain tumours: a symptom to reappraise critically. AB - Headache can be either a late or early symptom of a brain tumour, depending on the location of the tumour. A constant, progressively increasing pain, or a change in the character of headache pain, may alert the physician to this occurrence. Fortunately most people with headache, even persistent or severe headaches, do not have a tumour. In this work we review the literature about prevalence of headache as an isolated/early symptom of brain tumour and report our experience. PMID- 15549526 TI - Headache and inflammatory disorders of the central nervous system. AB - The subcommittee of the International Headache Society for headache classification (ICHD-II) has recently recognised that secondary headaches may occur in patients affected by inflammatory diseases (ID) of the central nervous system (CNS), classifying them among the headaches attributed to non-vascular intracranial disorders. The aim of the study was to verify the association between headache and inflammatory non-infectious diseases of the CNS, by a review of the literature data on the topic, integrated by personal cases and data. Secondary headaches may occur in four main disorders: neurosarcoidosis (sec 7.3.1), aseptic (non-infectious) meningitis (7.3.2), other non-infectious ID (7.3.3) and lymphocytic hypophysitis (7.3.4). Headache and/or primary headaches are frequently reported in patients with neurosarcoidosis (30%), Behcet's syndrome (BS) (55%) and acute disseminated encephalomyelitis (45-58%). Recent data show a high incidence of headache also in multiple sclerosis (MS) (58%) (not mentioned in ICHD-II). The association between headache and inflammatory dysimmune diseases of the CNS, in particular BS and MS, might suggest a pathogenetic relationship. PMID- 15549527 TI - Neuroradiological investigations in secondary headaches. AB - In this work, we examine the neuroradiologic features of the main non vascular clinical conditions responsible for secondary headache; excluding CSF hypotension, which will be treated extensively in another work in this supplement. Headache is not a constant feature of intracranial mass lesions, even of large extension. Headache has a high diagnostic value in children, as it can be the only heralding symptom, sometimes even for a long time, of severe intracranial pathologies, which later give rise to seizure or focal neurological signs. Particular attention should be paid to children affected by leukaemia under pharmacological treatment, in which headache is almost always the presenting symptom of serious neurological syndromes, consequent to antiblastic drugs. PMID- 15549528 TI - Tension-type headache and fibromyalgia: what's common, what's different? AB - Chronic tension-type headache and fibromyalgia have striking clinical and pathophysiologic similarities. They can be associated in patients. In both conditions there is evidence of altered processing of peripheral nociceptive information. Peripheral sensitisation of musculotendinous nociceptors may play a role and, at least in chronic tension-type headache, there are indications of central sensitisation. As for the clinical presentation, there also pathophysiological differences between the two disorders. A better understanding of both these differences and similarities may hopefully help in the future management of patients. PMID- 15549529 TI - Anticonvulsant drugs in primary headaches prophylaxis. AB - Anticonvulsant drugs have been used in migraine prophylaxis since 1970. In recent years, new antiepileptic medications have given rise to much interest in pain control. Primary headaches prophylaxis is still based on old drugs, and physicians facing these conditions are always prompted to use any new possible choice. Among primary headaches, the most studied drug over last 15 years was divalproex sodium, and many papers showed its efficacy in the treatment of migraine headaches. Valproate is well tolerated and many dosages have been used with success. For the newer drugs, such as gabapentin, lamotrigine or topiramate, evidence is less strong but has been rapidly increasing in the last 5 years. In particular, topiramate has much more evidence of a good efficacy and a safe profile. We review the principal characteristics of their use, according to dosages, lasting of treatments, side effects and significant efficacy. PMID- 15549530 TI - Strategies for the treatment of autonomic trigeminal cephalalgias. AB - Trigeminal autonomic cephalalgias (TACs) are a group of primary headache syndromes characterised by two main clinical characteristics: pain and oculofacial autonomic phenomena. Three headache forms are grouped as TACs: cluster headache (CH), paroxysmal hemicrania (PH) and short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT). These are distinguished mainly on the basis of attack duration. It lasts from 15 to 180 min in CH, from 2 to 30 min in PH and from 5 to 240 s in SUNCT. The most effective drug preventative in PH is indomethacin even if in few cases other non steroidal anti-inflammatory drugs have been reported to be effective. SUNCT is commonly described as drug resistant. Recent studies report that lamotrigin may be the drug of choice for SUNCT. PMID- 15549531 TI - Therapy of primary headaches: the role of antidepressants. AB - Antidepressants are included in evidence-based guidelines for the prophylactic therapy of migraine. Although they can cause several side effects depending on the neurochemical activity, and are to be used with caution in older patients, some of them have a well-documented efficacy. Amitriptyline is classified as a Group 1 drug, whereas Fluoxetine is included in Group 2. There is fair support for the effectiveness of other serotonine reuptake inhibitors in migraine prevention. Amitriptyline has demonstrated a consistent efficacy in Chronic Tension Type Headache, and Mirtazapine has a promising profile for the treatment of the same disease. PMID- 15549532 TI - Preventive migraine therapy: what is new. AB - In this paper we review new treatment options for migraine prevention. Because we focus on new drugs, some of the data we present herein were acquired in well design double blind, controlled studies, while the efficacy of other medication is supported only by open, uncontrolled trials (noted in the text wherever appropriate). PMID- 15549533 TI - Management of headache in emergency room. AB - Headache is one of the most common symptoms that leads patients to the emergency room (ER) and is often related to diseases requiring prompt diagnosis and immediate treatment. This consideration brought us to consider the importance of the neurologist in improving the management of patients arriving in the ER with headache. We carried out a study for testing the degree of agreement between ER physician and neurologist using patient evaluation at headache centre (HC) as the gold standard. One hundred and seventeen patients with idiopathic (78) or symptomatic (39) headache were evaluated by the ER physician, the ER neurologist and the HC expert. The ER physician and the HC expert reached a fair agreement (Kappa=0.40); the other two pairs reached a moderate agreement (Kappa=0.57-0.60). There was no significant difference in the agreement of the three evaluators in patients with impairment of daily living activities or aged over 40. The agreement between the ER physician and the neurologist was lower (Kappa=0.58), especially in patients with their first headache episode. Based on our results, patients seen at the ER for a headache episode can be fairly successfully managed by the ER physician, except those who present a first attack, for whom neurological consultation is needed. PMID- 15549534 TI - Guidelines for the management of headache in the emergency department. AB - Headache is a common problem in an emergency department. The main task of the emergency room (ER) doctor is to differentiate the primary forms from the secondary ones. Given that secondary headaches can manifest themselves in such a wide variety of ways, a differential diagnosis can then become extremely difficult. Guidelines can be found in the literature which help ER doctors with this task. Here we propose a procedure whereby four different clinical scenarios are defined for nontraumatic headache and then evidence-based suggestions are given for managing them in the emergency department. PMID- 15549535 TI - Headache in the emergency department. AB - Patients with headache commonly seek care at an emergency department (ED). Patients with headache in fact account for between 1 and 2% of admissions to an ED. Therefore the ED physician must recognize symptoms and characteristics of headache that signal a potential significant organic problem in order to select appropriate tests and treatment. Key to the correct management of headache in ED is a careful, thorough history of the patient. This article summarizes what the ED physician should take into consideration in order to distinguish who simply needs reassurance and analgesia and who needs further investigation. PMID- 15549536 TI - The headache in the Emergency Department. AB - The headache is a very frequent symptom and represents the 0.36%-2.5% of all reasons of claim to Emergency Department. Even if it is rarely related to high risk diseases, it is mandatory to promptly differentiate life-threatening conditions. In order to establish a correct diagnostic and therapeutic pathway and ask for aimed specialistic consultation, the emergency physician must be familiar with the various categories of headache. It is important to distinguish between essential headache and secondary headache. All patients presenting to the emergency department with the complaint of headache should be interviewed carefully regarding their history. The quality of pain associated with the intensity, location, rate, duration, modality of onset, relieving or worsening conditions, response to drugs, symptoms or signs associated must be investigated as well. Careful neurological examination including the vision of fundus oculi and the evaluation of rigor nucalis can provide further important diagnostic information. Laboratory exams do not usually give significant issues in the majority of patients with headache. However, dosage of inflammation index can be useful when an infective or inflammatory disease is suspected. CT scan can rule out the suspicion of organic intracranial causes. When the physician suspects meningitis or subarachnoid hemorrhage (SAH) not showed by CT scanning, rachicentesis can turn out diagnostic. The modality of onset, clinical characteristics and differential diagnosis of subarachnoid hemorrhage, intracranial hypertension, colloidal cyst of the third ventricle, trigeminal neuralgia, temporal arteritis and pituitary adenomas and apoplexy will be discussed. These diseases are not only of neurological and neurosurgical interest, but involve also the physician in the Emergency Department. PMID- 15549537 TI - Neurophysiological tests in primary headaches. AB - An impressive number of studies regarding the usefulness of neurophysiological procedures in primary headaches have been performed, but some of their results are difficult to interpret. However electrophysiological methods are useful to investigate peripheral and central mechanisms underlying this disorders. Taken globally, this results argue in favour of the neuronal pathogenesis of migraine as a final result of modified brainstem inputs to the cortex and/or consequence of a functional neuronal derangement. In this paper are briefly reviewed some of the principal data nowadays reported. PMID- 15549538 TI - Headache in cerebral venous thrombosis. AB - In the past, cerebral venous thrombosis (CVT) was considered a rare, devastating disease. The widespread use of angiography, magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) over the years has made early diagnosis of CVT possible and has completely changed the perception of this condition. CVT is much more common than previously thought. It has a wide spectrum of clinical presentation, multiple causes and unpredictable outcome. In contrast to arterial stroke, headache is the most frequent and, at times, the only symptom of CVT. It is crucial to recognise the association of headache with CVT in order to reach a correct diagnosis and to start the appropriate treatment as soon as possible. Therefore in order to define the headache features useful for an early diagnosis of CVT we reviewed the current literature on this topic and performed both a retrospective and a prospective study. In the literature we found that this headache has no specific features as it can be of any grade of severity and is slightly more frequently diffuse than localised. Its onset is usually subacute over a few days but it can also be acute or chronic. It is mostly persistent but can occasionally be intermittent. Headache attributed to CVT is sometimes misleading, mimicking migraine, subarachnoid haemorrhage, CSF hypertension or hypotension. However this information has been derived from case series which lacked a control group and thus have low statistical strength. Our retrospective study, confirmed by preliminary results in the prospective multicentric study, showed that headache in CVT is as often acute as subacute, and that it is more frequently localised than diffuse. Finally, both studies showed a significant correlation between headache of acute onset and severe intensity and CVT. We then conclude that these headache features, especially in the presence of underlying prothrombotic conditions, should lead clinicians to consider the diagnosis of CVT and to require appropriate neuroimaging examinations. PMID- 15549539 TI - Oral contraceptives in women with migraine: balancing risks and benefits. AB - Oral contraceptives (OCs) are a safe and highly effective method of birth control, but can also be associated with some risks, mainly a potential thrombotic risk. OCs may condition the course of headache and sometimes start it, but their influence on the clinical evolution of migraine is not easily assessable. The last Classification of Headache Disorders of the International Headache Society clearly identifies an "exogenous hormone-induced headache" that could be triggered by intake of OCs. Old high-dose OCs could effectively worsen headache in a significant proportion of patients, but the newest formulations influence headache course to a lesser extent. In any case, while an increase in migraine frequency or intensity do not oblige the cessation of OCs, experiencing a migraine aura for the first time, or even a clear worsening of a preexistent aura suggest discontinuation of OCs. Even if both migraine and OCs intake are associated with an increased risk of ischaemic stroke, migraine per se is not a contraindication for OCs use; however, patients suffering from migraine with aura generally show a greater thrombotic risk than women with migraine without aura. Other risk factors (patient's age, tobacco use, hypertension, hyperlipidaemia, obesity and diabetes) must be carefully considered when prescribing OCs in migraine patients. Furthermore, all OCs, even those with low oestrogen content, are a major risk for venous thrombosis, particularly in women with hereditary thrombophilia. A thorough laboratory control of the genetics of prothrombotic factors and coagulative parameters should precede any decision of OCs prescription in migraine patients. PMID- 15549540 TI - Sentinel headache. AB - Patients with subarachnoid haemorrhage (SAH) frequently describe the occurrence of an underestimated or even ignored severe headache in the days or weeks preceding the bleeding. If recognised early, this warning headache might lead to specific investigations and, if indicated, a surgical approach might avoid a dramatic haemorrhagic event. In a recent and exhaustive systematic review, the incidence of a sentinel headache (SH) was evaluated in a range of 10-43% of SAH patients. SH seems to be due to a minor bleeding from a leak of a berry aneurysm and usually occurs in the preceding two weeks. Such a period is similar to the one for rebleeding in SAH and supports the hypothesis of the warning leak. Nevertheless, a warning headache can precede a SAH in unruptured aneurysm even without a minor bleeding. Underestimation or misdiagnosis of SH depends on incorrect evaluation of the headache characteristics (unusual, severe, abrupt, thunderclap), overestimation of cranial CT sensitivity (false negative increasing over the elapsing time), failure to perform lumbar puncture (LP) in patients with negative CT, incorrect evaluation of CSF findings (xanthochromia may be absent in the first 12 h) and failure to differentiate traumatic tap from true SAH. Considering the diagnosis of SH in all cases of a severe, sudden-onset (thunderclap) headache, and performing all the appropriate diagnostic exams, including LP if necessary, could prevent subsequent massive bleeding and its invalidating or fatal consequences. PMID- 15549541 TI - Towards the computerisation of ANIRCEF Headache Centres. Presentation of AIDA CEFALEE, a computer assisted diagnosis database for the management of headache patients. AB - Because of the lack of biochemical or neuroradiological markers, the diagnostic assessment of primary headaches rests on a clinical history collected by an experienced physician; the articulated criteria of the International Headache Society (IHS) represent the only available tool to test objectively the diagnostic hypothesis rising from clinical interview. However, the complexity reached by IHS criteria still represents its major limit to extensive application, at least in common clinical settings. The use of modern information technology (IT), which is specifically designed to manage complex problems with a large number of variables, seems to be the best choice to counteract the complexity of IHS classification. In this paper AIDA CEFALEE, a user-friendly client-server database for the management of headache patients, is presented. The system integrates a computer assisted diagnosis module, which may help to extend the correct use of IHS diagnostic criteria to any clinical setting. The interoperability of the system may represent the possible infrastructure of a National Network of ANIRCEF Headache Centres. PMID- 15549542 TI - Headache in children and adolescents: conventional and unconventional approaches to treatment. AB - Headache is one of the most common conditions to affect children and adolescents in industrialised countries. Studies indicate a prevalence of 8% to 60%. In over 40% of migraineurs the condition begins before 18 years of age. For clinical researchers, headache in the young is of interest because its causes are easier to investigate than in older people, as the clinical history is brief, the condition has had no time to become chronic, and it is not accompanied by changes in pain neuromodulation and neurotransmission, or the development of a chronic pain/stress-related personality that is common in adults with longstanding headache. The wide variation in reported headache prevalence in young people may be ascribed to lack of representativity of studies and vagueness of diagnostic criteria in studies performed before 1988. The publication of the International Headache Society IHS classification in 1988 for the first time made it possible to accurately diagnose the various headache forms. The diagnostic criteria for adults were adopted for use in young patients. Since that time a burgeoning number of studies on headache in young people have been published. PMID- 15549543 TI - Update on menstrual migraine: from clinical aspects to therapeutical strategies. AB - Migraine occurrence is strongly influenced by the hormonal fluctuations of the female reproductive cycle; at least 60% of women affected by migraine relate the periodicity of their attacks to the menstrual cycle. The so-called menstrual migraine, which occurs immediately before, during or at the end of the menstrual flow, has been a largely undefined condition, including some clinical subtypes which are not well defined. In the last edition of the International Classification of Headache Disorders (ICHD-II), menstrual migraine gained new attention in the Appendix, where three clinical patterns were pointed out: pure menstrual migraine without aura; menstrually related migraine without aura and non-menstrual migraine without aura. Menstrual migraine attacks show severe intensity, long duration (lasting even more than 72 h), marked unresponsiveness to pharmacological treatments, and present higher recurrence rate and work related disability than non-menstrual attacks. The pharmacological treatment of menstrual migraine can require specific cyclic prophylactic approaches (non steroidal anti-inflammatory drugs, coxibs, magnesium, long half-life triptans or oestrogen supplements in various formulations), but usually the low frequency of attacks suggests a first approach with specific symptomatic drugs. Preference should be given to triptans, due to their specificity in controlling migraine pain and its accompanying symptomatology; among them, in particular for sumatriptan, many specific studies proved a real effectiveness in the management of acute menstrual migraine attack. PMID- 15549544 TI - Primary headaches in children and adolescents. AB - Headache is one of the commonest conditions to affect children and adolescents in industrialised countries. Studies indicate a prevalence of 8-60%. In over 40% of migraineurs the condition begins before 18 years of age. For clinical researchers, headache in the young is of interest because its causes are easier to investigate than in older people, as the clinical history is brief, the condition has had no time to become chronic, and it is not accompanied by changes in pain neuromodulation and neurotransmission, or the development of a chronic pain/stress-related personality that is common in adults with longstanding headache. The wide variation in reported headache prevalence in young people may be ascribed to lack of representativity of studies and vagueness of diagnostic criteria in studies performed before 1988. The publication of the International Headache Society IHS classification in 1988 for the first time made it possible to accurately diagnose the various headache forms. The diagnostic criteria for adults were adopted for use in young patients. Since that time a burgeoning number of studies on headache in young people have been published. PMID- 15549545 TI - An updated review of migraine and co-morbid psychiatric disorders. AB - General population studies suggest a non-casual association (co-morbidity) between migraine, major depression and anxiety disorders (panic attack disorder, obsessive-compulsive disorder, generalised anxiety disorder). The risk of developing affective and anxiety disorders has not increased uniformly in the different migraine subtypes, but it is more elevated in migraine with aura patients. The relationship between migraine and depression is "bi-directional" (i. e., migraineurs have a more than three-fold risk of developing depression compared with non-migraine patients, while depression patients that have never suffered from migraine before have a more than three-fold risk of developing migraine compared with non-depressed patients) and specific (i. e., the presence of migraine or severe non-migraine headache increases a patient's risk of developing depression or panic attack disorder, whereas the presence of depression or panic attack disorder is associated with a greater risk of developing migraine, but not severe non-migraine headache). The association described in this review has significant pathogenetic, clinical and therapeutical implications. PMID- 15549546 TI - Pathophysiology of migraine. AB - The exact pathogenesis of migraine remains to be determined. In particular there is increasing evidence for the neural basis of migraine. We now have a body of data supporting the concept of central neuronal hyperexcitability as a pivotal physiological disturbance predisposing to migraine. The reasons for increased neuronal excitability may be multifactorial. Most recently, abnormality of calcium channels has been introduced as a potential mechanism of interictal neuronal excitability. Mutant voltage gated P/Q type calcium channel genes likely influence presynaptic neurotransmitter release, possibly of excitatory amino-acid systems or inhibitory. It could therefore be hypothesised that genetic abnormalities result in a lowered threshold of response to trigger factors. There is also evidence from spectroscopic studies that magnesium is low in migraine. We currently conceive of a migraine attack as originating in the brain. Triggers of an attack initiate a depolarising neuroelectric and metabolic event likened to the spreading depression of Leao. This event activates the headache and associated features of the attack by mechanisms that remain to be determined, but appear to involve either peripheral trigeminovascular or brain stem pathways, or both. Excitability of cell membranes, perhaps in part genetically determined, is the brain's route of susceptibility to attacks. Factors that increase or decrease neuronal excitability constitute the threshold for triggering attacks. PMID- 15549547 TI - Treatment strategies in migraine patients. AB - The goal of migraine treatment is to alleviate the symptoms of acute attacks and improve patients' quality of life. Therapeutic options and strategies principally rely on three types of approach: correction of causative factors; acute treatment of attacks; and prophylaxis. The quality of evidence supporting efficacy, personal experience, tolerability and safety profiles must guide the choice of a particular medication; nonetheless, we ought to keep in mind that therapeutic options should also be customised to target the individual patient, both in terms of personal characteristics and underlying comorbidities. Also, the framework of information the patient is given represents an essential component of migraine management, along with his/her active involvement in the therapeutic program and schedule. PMID- 15549548 TI - Topiramate in migraine prevention: evidence-based medicine from clinical trials. AB - The importance of evidence-based medicine is often underestimated when migraine preventive treatments are chosen. Evidence from large, well-conducted, placebo controlled trials of sufficient duration should be given particular consideration when evaluating the efficacy, tolerability and safety of a medication for a broad range of patients. Pizotifen, propranolol, flunarizine, amitriptyline, divalproex sodium and topiramate have all been evaluated for efficacy in migraine prevention in double-blind, placebo-controlled trials. The largest clinical programme in migraine prevention, studying these or any other agents, was comprised of 2 pivotal trials, as well as other studies, evaluating topiramate for preventive therapy in migraine. Results from these trials indicated that topiramate (100 mg/day) has proven safety and efficacy in migraine prevention. Response rates were high, and onset of action usually occurred within the first month of treatment. Therefore, topiramate has demonstrated safety and represents an effective option in the prevention of migraine, as supported by extensive clinical research. PMID- 15549549 TI - Measure of negative impact of migraine on daily activities, social relationships and therapeutic approach. AB - The study has been conducted in 9 European countries, interviewing 200 women in each country, aged 18-35, in fully working or studying period, to get a total of 1810 people. Migraine or severe headache was recognised and patients were studied to understand their behaviour. The features and severity of headache, the use of different drugs, the relationship between physicians and patients, the disability during attacks, the psychological aspects and the feeling of impotence that migraine patients experienced during their lives, are analysed and reported. PMID- 15549550 TI - Workplace disability in migraine: an Italian experience. AB - Workplace disability due to migraine has not been extensively researched in non English speaking countries. We assessed the repercussions of headache, and particularly of migraine, on work in a sample of employees from an Italian company (Bulgari). Information was obtained through a self-answering questionnaire in "all headaches" sufferers, and through direct interview in migraine sufferers (diagnosis according to IHS criteria). Headache frequency, pain intensity and headache-related disability were higher in migraineurs than in "all headaches" sufferers. About a quarter of migraineurs missed at least one day in the three months prior to the interview due to headache, and around 10% lost two or more days over the same period. Moore than 50% of migraineurs reported 1-7 days per month at work with headache, with reduction in productivity level by 50% or more in 15% of respondents. Our data confirmed that headaches, and particularly migraine, cause a considerable reduction in workplace productivity. Workplace interventions to effectively manage migraine are needed. PMID- 15549551 TI - Headache screening and diagnosis. AB - In most cases, diagnosis of the various headache subtypes is possible through the accurate collection of medical history data. However, sometimes serious problems of differential diagnosis may be encountered. Therefore, the distinction between migraine without aura and tension-type headache is not always easy, the relationship between chronic migraine and medication overuse headache is a complex one, and differentiation of chronic tension-type headache vs. new daily persistent headache is often problematic. A clear knowledge of the distinctive clinical features of the various headache subtypes is necessary to establish a correct diagnosis in the group of unilateral headaches with short-lived attacks and in the group of headaches with nocturnal onset. PMID- 15549552 TI - ID-migraine. AB - Migraine is a highly prevalent and disabling illness that remains substantially undiagnosed in primary care. Validated migraine screening tools are of potential value in addressing this important barrier for migraine treatment. Herein we briefly discuss reasons for screening migraine in the primary care setting, and focus on the ID-migraine, a validated migraine screening tool. PMID- 15549553 TI - MIDAS questionnaire modification for a new MIDAS junior questionnaire: a clinical experience at the Neurological Institute "C. Besta". AB - During the last decade researchers have begun to employ standardised methodologies to investigate the global impact of primary headaches. Disease specific instruments have been developed to measure headache-related disability. The MIDAS questionnaire, which is the most extensively studied of these instruments, was designed to assess the overall impact of headaches over the 3 months before compilation. The MIDAS questionnaire is an optimal tool to assess headache-related disability in adults in relation to patients' daily activities. Primary headaches are a recurrent problem for children and adolescents. Forty percent of children have experienced headaches by the age of 7 years increasing to 75% by the age of 15. In a recent report we determined the suitability of the MIDAS questionnaire in its original form for assessing disability in children and adolescents suffering from different kinds of headache. This was the first step of a line of research aimed to develop a new MIDAS questionnaire adapted for young patients. In this second study the aims were: (1) to produce a new version of the MIDAS questionnaire specific for young patients suffering from different forms of headache; (2) to assess the reliability of this new instrument; (3) to assess its sensitivity to treatment intervention. PMID- 15549554 TI - The role of neuroimaging in the diagnosis of headache in childhood and adolescence: a multicentre study. AB - Following the diagnostic indications of the Guidelines for the diagnosis and therapy of juvenile headache, we present the results of a prospective, multicentre study of headache patients aimed at evaluating the utility of neuroradiologic testing in the diagnosis of headache. A total of 6535 subjects up to age 18 were studied, and 1485 underwent neuroimaging testing based on the indications of the diagnostic Flow-Chart. Abnormal results were observed in 273 (18.5%) subjects. Incidental findings were observed in 138 (9.3%) subjects, not correlated with the headache pathology, whereas alterations that led to the diagnosis of secondary headache were observed in 135 (9.1%) subjects. In conclusion, our data support a greater incidence of positive neuroimaging examinations among patients who underwent testing based on suspicion of a secondary headache pathology when compared with the low percentages observed in study populations. PMID- 15549555 TI - Picotamide in migraine aura prevention: a pilot study. AB - In an open, preliminary trial we evaluated the use of picotamide, an antiplatelet drug, in the prophylactic treatment of migraine aura (MA). Twenty-two women suffering from migraine with typical aura or MA without headache, diagnosed according to International Headache Society criteria, entered a nine-month study. They underwent a three-month run-in period free of prophylactic drugs, followed by a six-month treatment period (subdivided in two trimesters, TI and TII) with 300 mg of picotamide administered twice daily. A detailed diary reporting neurological symptoms, duration and frequency of MA was compiled by patients along the trial time. The number of MA significantly decreased during treatment (from 6.85+/-3.82 in the run-in trimester to 2.85+/-2.72 during TI and to 2.55+/ 2.89 during TII). Also, MA duration was decreased significantly, being 36.75+/ 20.28 min during run-in, 20.00+/-16.94 during TI and 17.75+/-16.26 during TII. In 25% of patients MA totally disappeared. The number and the features of aura neurological symptoms were also positively modified by the use of picotamide. No serious adverse event was provoked by picotamide administration. Picotamide is effective in reducing MA frequency, duration and symptomatology. The effect is clearly evident in the first trimester of treatment and is maintained with no further modifications during the second trimester. PMID- 15549556 TI - Pharmacological behavioural treatment for children and adolescents with tension type headache: preliminary data. AB - Recurrent headaches are a significant health problem for young patients. Most current investigations have employed limited modalities (either medication or behavioural) and few have included treatment comparisons. The purpose of this study was to compare relaxation training (provided in a limited contact format) and amitriptyline in the treatment of young tension-type headache (TTH) sufferers. Follow-up is planned for 3, 6 and 12 months, at which time patients complete headache logs and an Italian version of the Migraine Disability Assessment (MIDAS) Questionnaire that has been adapted for this age group. Two groups of TTH sufferers of similar age and characteristics were studied. Clinical results, MIDAS total score, and individual values for items A and B were collected at the first follow-up (3 months). The clinical improvement is significant for both groups. Although the clinical results are similar in both groups, relaxation therapy seems to be more accepted than pharmacological therapy. These data, however, are preliminary and the sample sizes are small, so these conclusions are tentative. We will continue our data collection for 12 months. PMID- 15549557 TI - Chronic migraine with medication overuse: treatment outcome and disability at 3 years follow-up. AB - Patients with chronic migraine and medication overuse are particularly difficult to treat. No clear consensus exists about treatment strategies to be used and little data exists about the functional impact of headache in these patients. The purpose of the study was to determine (1) the clinical course of a sample of chronic migraine patients with medication overuse 36 months following treatment intervention and (2) whether functional impairment, assessed by the Migraine Disability Assessment (MIDAS) questionnaire, improved upon treatment. Of 106 patients meeting the criteria for chronic migraine with medication overuse (according to Silberstein and Lipton), 71 went on to complete a structured inpatient treatment, consisting of medication withdrawal and then prophylactic treatment. As a group, the patients were significantly improved at 36-month follow-up, with respect to 2 headache parameters (days of headache per month and number of used medications per month assessed by the diary card) and 2 measures of functional impact extracted from the MIDAS questionnaire (MIDAS total score and frequency of headache). Chronic migraine accompanied with medication overuse led to considerable disability prior to treatment. However, notable improvement both in headache parameters and in disability measures occurred concurrently with treatment. This suggests that successful treatment has more wide-ranging positive benefits beyond mere symptom reduction. To our knowledge, this is the first investigation where the MIDAS questionnaire has been used as an outcome measure in patients with chronic headache to assess disability during such a long follow up period. PMID- 15549558 TI - MIDAS questionnaire in the emergency setting. AB - Migraine is a common disorder and is a major cause of disability and loss of working performance in western countries. Therefore, many tools have been developed to assess migraine related disability. Among these, the Migraine Disability Assessment (MIDAS) questionnaire has been shown to be of particular interest, as it is valid, reliable and useful for therapeutic decisions. In this pilot study, we address the validity of the MIDAS questionnaire in an unselected population of migraine patients in the emergency setting. We found that the MIDAS scores in the emergency room were similar to those collected in a specialised headache centre. This result suggests that the MIDAS questionnaire could be reliably used in the emergency setting, hence avoiding unnecessary delays in the treatment of migraine patients. PMID- 15549559 TI - Identifying patients who require a change in their current acute migraine treatment: the Migraine Assessment of Current Therapy (Migraine-ACT) questionnaire. AB - The aim of the study was to design and test a new, easy to use, assessment tool, the Migraine Assessment of Current Therapy (Migraine-ACT), for identifying patients who require a change in their acute treatment. A 27-item questionnaire was developed by an international advisory board including questions formulated in four domains: headache impact, global assessment of relief, consistency of response and emotional response. Migraine patients entered a multinational, prospective study to investigate the test-retest reliability and construct validity of the tool, which was completed by the patients on two occasions. Test retest reliability was assessed by Pearson's and by Spearman correlation coefficients. Construct validity was assessed by correlating patients' answers to the 27-item questionnaire with those of well-reported measures: SF-36, MIDAS and Migraine Therapy Assessment Questionnaire (MTAQ). The test-retest reliability of the 27 initial questions ranged from good to excellent. Correlations of all items with SF-36, MIDAS and MTAQ scores--assessed by discriminatory t-tests--indicated that the following 4 were the most discriminating items: Does your migraine medication work consistently, in the majority of your attacks? Does the headache pain disappear within 2 hours? Are you able to function normally within 2 hours? Are you comfortable enough with your medication to be able to plan your daily activities? The 4-item Migraine-ACT is a brief, simple, and reliable assessment tool to identify patients who require a change in their acute migraine treatment, and can be recommended for primary care physicians, neurologists and headache clinicians. PMID- 15549560 TI - Elusive amines and cluster headache: mutational analysis of trace amine receptor cluster on chromosome 6q23. AB - Cluster headache (CH) is characterised by unilateral pain and ipsilateral autonomic features. To date, no molecular genetic evidence has been shown for CH. Small pedigrees and low penetrance render the identification of the CH-gene quite difficult. Nonetheless the association of CH and migraine to a new class of amine, namely trace or elusive amines such as tyramine, octopamine and synephrine, has recently been demonstrated. In particular, in comparison to healthy control subjects, all these neurotransmitters have been found to be greatly elevated in CH sufferers in plasma and platelets both in active and remission periods. A cluster of gene-encoding G-protein-coupled receptors that bind and are activated by trace amines was identified in the long arm of chromosome 6q23. We evaluated two families with CH by linkage analysis to 6q23 region and the mutation scanning of the TAR 1, TAR 3, TAR 4, TAR 5, PNR and GPR58 genes by denaturing high liquid chromatography is in progress in 16 familial cases. PMID- 15549561 TI - New daily persistent headache: clinical and serological characteristics in a retrospective study. AB - We present a retrospective clinical study of 18 cases of new daily persistent headache (NDPH), a rare chronic headache, included in the fourth chapter of the II IHS classification; the pathophysiology of NDPH is unknown but a link with viral infections (especially Epstein-Barr virus (EBV)) has been suggested. Comparing our series with the other two published until now, we did not find any particular difference, as regards to clinical aspects. However, our laboratory tests show a recent herpes simplex virus infection in 42% and cytomegalovirus in 11% of cases; moreover we could not find any EBV infection. Our data suggest that viruses other than EBV can play a role in NDPH. PMID- 15549562 TI - Visually evoked phase synchronisation changes of alpha rhythm in migraine. Correlations with clinical features. AB - We investigate phase synchronisation in EEG recordings from migraine patients. We use the analytic signal technique, based on the Hilbert transform, and find that migraine brains are characterised by enhanced alpha band phase synchronisation in the presence of visual stimuli. In migraine, the brain synchronises to the idling rhythm of the visual areas under certain photic stimulations; hypersynchronisation of the alpha rhythm may suggest a state of cortical hypoexcitability during the interictal phase of migraine. PMID- 15549563 TI - Cortical function abnormalities in migraine: neurophysiological and neuropsychological evidence from reaction times and event-related potentials to the Stroop test. AB - Cortical hyperexcitability was studied in migraine patients using reaction times (RT's) and Event-Related Potentials (ERP) to the Stroop test. We found a slower RTs in patients if compared to controls, supporting the hypothesis of a mild cortical functions impairment even in interictal periods in this group of patients. PMID- 15549564 TI - Visual evoked potentials in migraine. AB - Migraine is a chronic disorder. Visual symptoms and hypersensitivity to light stimuli are common. The aim of this study is the analysis of visual system in migraineurs by visual evoked potentials (VEP). We studied 53 migraineurs (21 with prophylactic migraine treatment and 32 without preventive therapy) and 20 healthy control subjects. We found lower P100 latencies in migraineurs without therapy compared to controls. In treated patients, P100 latencies showed the same trend seen in the control group. We speculate a different responsiveness of the visual system in migraineurs probably due to a dysmodulation of sensor input leading to facilitation of visual processing. PMID- 15549565 TI - Migraine, daily chronic headache and fibromyalgia in the same patient: an evolutive "continuum" of non organic chronic pain? About 100 clinical cases. AB - Aim of this study is to evaluate if migraine, daily chronic headache and fibromyalgia in the same patient can be considered as an evolutive continuum of non organic chronic pain. Therefore, migraine, daily chronic headache and fibromyalgia should be considered the expression of chronic antinociceptive system alteration. PMID- 15549566 TI - Spontaneous cerebrospinal fluid leak syndrome: report of 18 cases. AB - We examined a group of 18 consecutive patients with spontaneous cerebrospinal fluid leak syndrome (SCSFLS) and investigated clinical, MRI, radioisotope findings and therapeutic outcome of this syndrome. PMID- 15549567 TI - Optic neuritis: diagnostic criteria application in clinical practice. AB - Optic neuritis (ON) refers to any inflammatory optic neuropathy. In clinical practice ON is mainly diagnosed by ophthalmologists and less frequently by neurologists. ON diagnostic criteria are included in the Classification of International Headache Society (IHS) and in other classification systems, both in neurological and ophthalmologic fields. The aims of this study were to verify the application of IHS ON diagnostic criteria in clinical practice and the role of the ocular pain qualitative aspects. We performed a partially retrospective (140 cases) and prospective (43 cases) study analysing the clinical characteristics of patients with ON. We observed retro orbital pain in a huge percentage of patients; it was provoked or spontaneous and worsened by eye movements. We found that the new IHS classification criteria (IHS 2004) do not fully satisfy the requirements for ON diagnosis. Further study is necessary to validate the diagnostic criteria of ON in clinical practice. PMID- 15549568 TI - Does headache represent a clinical marker in early diagnosis of cerebral venous thrombosis? A prospective multicentric study. AB - The main aim of this study is to look for early clinical markers of cerebral venous thrombosis (CVT). As headache represents the major clinical manifestation at presentation we focused our attention on this symptom. We present the preliminary results of a prospective multicentric study that includes cases diagnosed as CVT in the participating centres. We have so far studied 35 patients (5 males and 30 females) from the ages of 18 to 78. The most frequent manifestation was headache (77.1%). It was more frequently localised (66.7%) and continuous (77.8%). The onset of pain was mostly acute-subacute (38.5%-50.0%) and the intensity moderate-severe (37.0%-51.9%). On univariate analysis, we found a positive correlation between CVT, acute headache onset (p=0.001) and severe headache (p=0.004). These preliminary results seem in accordance with our previous findings in the retrospective study, suggesting that CVT is more often associated with acute-onset headache of severe intensity. PMID- 15549569 TI - Migrainous cerebral infarction: case reports. AB - Migraine is a common and chronic disorder. It is considered benign but several studies have suggested it as a rare risk factor for ischaemic stroke. The association is still conflicting and seems to be restricted to particular subgroups of patients (i.e., women under the age of 45, with migraine with aura, and particularly ones who smoke and use oral contraceptives). The pathogenetic mechanisms underlying this condition are not known. We describe 6 cases of migrainous stroke fully meeting the diagnostic criteria of the International Headache Society (IHS). For each patient, demographic and anamnestic data, clinical features, results of laboratory tests and neuroimaging findings were recorded. Five of the 6 cases were women (median age of 29, range from 23 to 40). The man was 36. All patients fulfilled the IHS criteria for migraine with aura. At the time of the event, 2 patients were taking oral contraceptives and smoked, one patient smoked and three patients had no vascular risk factors. The stroke manifested as homonymous hemianopia in 3 patients, lower homonymous quadrantopia in 1 patient and sensory symptom in 1 patient. The neurological examination was normal in 1 case. All patients underwent several tests with negative results: blood test (antithrombin III, protein C or S, autoantibodies), transthoracic and transoesophageal echocardiography, extracranial and intracranial Doppler sonography, and angiography which was not performed in 1 patient. All patients had a cerebral infarct visible on neuroimaging study (MRI): posterior cerebral artery in 4, middle cerebral artery in 1 and anterior cerebral artery in 1. We support the findings reported by others that migrainous stroke is more common in young women affected by migraine with aura. In consideration of the high prevalence of migraine in the population, further research is indicated and necessary to establish if migraine is independent from other vascular risk factors. PMID- 15549571 TI - Headache and neurology. AB - The aim of this lecture is to analyse the position of headaches and especially of migraine within the body of neurological knowledge. Historical, clinical and pathophysiological data have been selected for discussion. PMID- 15549572 TI - Headache classification: criticism and suggestions. AB - The International Classification of Headache Disorders 2nd Edition (ICHD-II), published in 2004, marks an unquestionable progress from the preceding 1988 edition, but the in-depth analysis it offers is not immune from drawbacks and shortcomings. First of all, it is still basically a classification of attacks and not of syndromes. For the migraine group, while the revised classification more accurately characterises migraine with aura, it fails to provide a sufficiently structured description of those forms of migraine without aura that over the years evolve to so-called daily chronic forms. These forms are not adequately recognised as chronic migraine, which ICHD-II includes among the complications of migraine. The inclusion of short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing (SUNCT) in the cluster headache group is bound to generate some perplexity, while the recognition of new daily persistent headache (NDPH) included in the group of other primary headaches as a separate clinical entity appears somewhat premature. Doubts are also raised by the actual existence of triptan-overuse headache, which ICHD-II includes in Group 8 among medication-overuse headaches. Finally, the addition of headache attributed to psychiatric disorder, which is certainly a good option in perspective, is not yet supported by an adequate systematisation. PMID- 15549573 TI - Methodology of studies on the epidemiology of headache. AB - Headache is a common clinical condition. A correct evaluation of its frequency, characteristics and aetiology is a prerequisite for health care planning and for the implementation of preventive and treatment measures. Descriptive and analytic epidemiological studies are the basis to improve the knowledge of the burden of headache in the community. However, the results of the epidemiological studies are heavily dependent on the quality of the design, the correctness of the conduct and the appropriateness of the data analysis. A proper knowledge of the problems in the conduction and interpretation of the epidemiological studies helps the investigators to optimise the study protocols and the practicing physicians to critically view the published results. PMID- 15549574 TI - Trigeminal autonomic cephalalgias: from pathophysiology to clinical aspects. AB - The strictly unilateral headaches, more commonly known as trigeminal autonomic cephalalgias (TACs), are characterised by severe, strictly unilateral pain in the territory of the distribution of the trigeminal nerve, associated with autonomic manifestations. The recent International Headache Society classification lists the strictly unilateral headaches as cluster headache (CH), episodic and chronic paroxysmal hemicrania, and short-lasting unilateral neuralgiform headache attacks with conjunctival injection and tearing. CH is the most common and best-defined of the TACs, whose pathophysiologies have not been adequately defined. Convincing proposals for pathophysiologic mechanisms must explain the trigeminal distribution of the pain, the homolateral autonomic manifestations; and, for CH, the usually periodic recurrence of the crises and clusters. With regard to CH, (i) the pain is always located periorbitally-frontally, implicating nociceptive mechanisms involving the trigeminal nerve; (ii) the autonomic manifestations homolateral to the pain seem to be both parasympathetic (lacrimation and rhinorrhoea) and sympathetic (ptosis and miosis); and (iii) the periodicity of the attacks and seasonal recurrence of the cluster periods suggest involvement of a biological clock within hypothalamus. PMID- 15549575 TI - Rare primary headaches: clinical insights. AB - So-called "rare" headaches, whose prevalence rate is lower than 1% or is not known at all and have been reported in only a few dozen cases to date, constitute a very heterogeneous group. Those that are best characterised from the clinical point of view can be classified into forms with prominent autonomic features and forms with sparse or no autonomic features. Among the former are trigeminal autonomic cephalalgias (TACs) and hemicrania continua, while the latter comprise classical trigeminal neuralgia, hypnic headache, primary thunderclap headache, and exploding head syndrome. The major clinical discriminating factor for the differential diagnosis of TACs is the relationship between duration and frequency of attacks: the forms in which pain is shorter lived are those with the higher frequency of daily attacks. Other aspects to be considered are the time pattern of symptoms, intensity and timing of attacks, the patient's behaviour during the attacks, the presence of any triggering factors and of the refractory period after an induced attack, and response to therapy, especially with indomethacin. Often these are little known clinical entities, which are not easily detected in clinical practice. For some of them, e. g., thunderclap headache, it is always necessary to perform instrumental tests to exclude the presence of underlying organic diseases. PMID- 15549576 TI - The contribution of functional neuroimaging to primary headaches. AB - Functional imaging techniques have begun to provide considerable insight into the pathophysiology of primary headache syndromes. PET and f-MRI have allowed to to monitor the physiological cortical reaction and nociceptor transmission of head pain, but more importantly have identified pathophysiological abnormalities and even the "motor" in migraine and cluster headache attacks. This has even prompted new treatment options such as DBS in cluster headache and will undoubtly change the way we see headache. Innovative techniques such as voxel- and deformation based morphometry have just started to unravel the structural consequences of chronic pain. Functional imaging will undoubtedly provide further opportunities to study and compare metabolic, haemodynamic and structural parameters in headache sufferers' brains. PMID- 15549577 TI - Contributions of biochemistry to the pathogenesis of primary headaches. AB - We briefly summarise biochemical anomalies of serotonin, norepinephrine, glutamic and aspartic acids, the main neurotransmitters of inhibitory and excitatory neuronal circuitries, found in primary headaches and their relationship with pathogenesis of migraine and cluster headache (CH). In addition, the high levels of circulating tyramine, octopamine and synephrine (elusive amines), recently reported in both migraine types and CH, are discussed in relation to the other "classic" amines findings. In particular it is suggested how abnormal levels of elusive amines may participate in the pathophysiology of migraine and CH acting through their specific trace amine receptors and alpha and beta receptors. The possible hypothesis that emerges from the analysis of these biochemical findings is that an imbalance of systems, with opposite neurophysiological functions related to the pain and other yet unknown functions, may constitute the biochemical phenotype of migraine with and without aura, and CH. PMID- 15549578 TI - The physiopathology of migraine: the contribution of genetics. AB - Recent advances in the studies of the genetic liability to migraine include the discovery of two genes responsible for familial hemiplegic migraine (FHM) and the analysis of several sites of linkage or genetic association for the so-called typical migraines, e. g., migraine with (MA) and without aura (MO). The 2 genes implicated in the genetics of FHM are CACNA1A for FHM1 and ATP1A2 for FHM2. It is still unclear how dysfunction in these genes may trigger attacks of migraine with hemiplegic features and, in at least part of the families with FHM, also paroxysmal or progressive ataxia and epileptic seizures. It appears that mutations in CACNA1A responsible for FHM1 alter calcium influx and calcium currents in neurons, possible factors of spreading depression like events. On the other hand, abnormal regulation of intracellular calcium concentrations could alter neurotransmitter release and other cellular functions. In the case of ATP1A2 mutations, haplo-insufficiency of the gene has been hypothesised to result in abnormal potassium level regulation because of faulty Na/K exchange with subsequent depolarisation and increased liability to spreading depression, or/and in abnormal calcium levels because of the concomitant activation of the Na/Ca exchanger, with a mechanism therefore comparable to that at work in FHM1. Much more work is clearly necessary to elucidate these pathophysiological mechanisms; advances in genetics however may represent important steps in the clarification of the physiopathology of the migraine attack. PMID- 15549579 TI - Research developments in the physiopathology of primary headaches. PMID- 15549580 TI - Photohemolysis of erythrocytes by He-Ne laser irradiation: the effect of power density. AB - The effects of He-Ne laser irradiation on erythrocyte hemolysis were studied. Erythrocytes are prone to be easily damaged with hemolysis under laser irradiation. It was found that the power density (PD) of laser irradiation, but not the total irradiation power, is the crucial parameter for the hemolysis. Under irradiation with a PD of 5.41 mW/microm(2), erythrocytes were photohemolyzed within a few seconds, while at a PD of 0.52 mW/microm(2) the average tolerance duration (TD; from the start of irradiation to hemolysis) of erythrocytes was 110 s. The TDs of erythrocytes were also individual erythrocyte dependent, which may be due to their different ages. The average TDs of erythrocyte samples from different donors were different, reflecting individual variations. PMID- 15549582 TI - Role of surgery in multidisciplinary treatment for solid cancers. AB - In the evolution of solid cancer, there are four steps: noninvasive tumor, local invasive cancer without metastasis, local invasive cancer with lymph node metastasis, and eventually systemic disease. For the first three phases, local treatment, including lymph node dissection, may cure the disease. The choice of local treatment depends on the tumor characteristics, but surgery remains important in many of these cancers. Gastric cancer is one of the typical tumors which remain locally invasive, with or without nodal metastasis, but without systemic metastasis for a rather long period. Metastasis to lymph nodes occurs, frequently even in T1 tumors, but seldom to other sites until the late stage. Thus, the target of local control is the regional lymph nodes. The Intergroup study IT-0116 proved the effect of chemoradiotherapy (CRT) for curable gastric cancer, and thus proved the insufficiency of limited surgery (D0/1). The conventional method of local control for gastric cancer is surgery, including regional lymph node dissection (D2). However, the superiority of D2 has not been proven by randomized controlled trials (RCTs). But all RCTs so far have a crucial problem in the quality of treatment given in the D2 arm. D2 is not a dangerous procedure if done by specialists in large-volume hospitals. D0/1 plus CRT is better than D0/1 alone, but it may be worse than D2 alone. The survival benefit of CRT after D2 is an open question. Establishing standard adjuvant chemotherapy after D2 is a more urgent clinical issue, and there is no reason to abandon D2 gastrectomy for curable gastric cancer in Japan. PMID- 15549583 TI - Stereotactic body radiation therapy for early-stage non-small-cell lung cancer: the Japanese experience. AB - Stereotactic body radiation therapy is a new treatment modality for early-stage non-small-cell lung cancer, and is being intensively investigated in the United States, the European Union, and Japan. We started a feasibility study of this therapy in July 1998, using a stereotactic body frame. The eligibility criteria for primary lung cancer were: (1) solitary tumor less than 4 cm; (2) inoperable, or the patient refused operation; (3) histologically confirmed malignancy; (4) no necessity for oxygen support; (5) performance status equal to or less than 2, and (6) the tumor was not close to the spinal cord. A total dose of 48 Gy was delivered in four fractions in 2 weeks in most patients. Lung toxicity was minimal. No grade II toxicities for spinal cord, bronchus, pulmonary artery, or esophagus were observed. Overall survival for 29 patients with stage IA, and 14 patients with stage IB disease was 87 % and 80 %, respectively. No local recurrence was observed in a follow-up of 3-50 months. Regional lymph node recurrence developed in 1 patient, and distant metastases developed in 4 patients. We retrospectively analyzed 241 patients from 13 Japanese institutions. The local recurrence rate was 20% when the biological equivalent dose (BED) was less than 100 Gy, and 6.5% when the BED was over 100 Gy. Overall survival at 3 years was 42% when the BED was less than 100 Gy, and 46% when it was over 100 Gy. In tumors which received a BED of more than 100 Gy, overall survival at 3 years was 91% for operable patients, and 50% for inoperable patients. Long-term results, in terms of local control, regional recurrence, survival, and complications, are not yet evaluated. However, this treatment modality is highly expected to be a standard treatment for inoperable patients, and it may be an alternative to lobectomy for operative patients. A prospective trial, which is now ongoing, will, answer these questions. PMID- 15549584 TI - Multidisciplinary approach to cancer treatment: a model for breast cancer treatment at the M.D. Anderson Cancer Center. AB - Despite that fact that a multidisciplinary approach is important for cancer treatment, this approach is not widely used in Japan. The M.D. Anderson Cancer Center (MDACC) is one of the most well-established cancer institutes in the world, and has implemented a unique multidisciplinary team management approach for the treatment of breast cancer. The efforts of MDACC to eliminate cancer have been ongoing for more than 60 years. Here, we describe the multidisciplinary approach used at MDACC for the treatment for breast cancer. We focus on the background of the institute, in terms of establishing its treatment model and educational system, and compare its multidisciplinary approach with the current approach used in Japan, in the hopes of influencing future directions in cancer therapy in Japan. PMID- 15549585 TI - Phase I/II study of irinotecan, 5-fluorouracil, and l-leucovorin combination therapy (modified Saltz regimen) in patients with metastatic colorectal cancer. AB - BACKGROUND: A combination of irinotecan 125 mg/m2, 5-fluorouracil (5-FU) 500 mg/m2, and leucovorin (LV) 20 mg/m2 (Saltz regimen; treatment on days 1, 8, 15, and 22 every 6 weeks) is widely used for the treatment of metastatic colorectal cancer. A modified schedule with chemotherapy on days 1 and 8 of a 21-day cycle was recommended in 2001 because of early treatment-related mortality. We conducted a phase I/II study of this modified Saltz regimen as first-line therapy in Japanese patients with metastatic colorectal cancer to assess the maximum tolerated dose (MTD) and the recommended dose of 5-FU when given with fixed doses of l-LV and irinotecan, and to evaluate the efficacy and the feasibility of this regimen. METHODS: Irinotecan, 5-FU, and l-LV were administered on days 1 and 8 of a 21-day cycle. Irinotecan 100 mg/m2 was given intravenously over the course of 90 min on day 1, followed by l-LV 10 mg/m2, and then 5-FU. The dose of 5-FU was escalated from 400 mg/m2 (level 1) to 500 mg/m2 (level 2). If neither level met the criteria for the MTD, the recommended dose was defined as level 2, and dose escalation was discontinued, because the maximum approved weekly dose of irinotecan alone in Japan is 100 mg/m2 and the dose of 5-FU in the original Saltz regimen was 500 mg/m2. RESULTS: One patient had grade 4 neutropenia with fever at level 1, and four patients had grade 3 neutropenia at level 2. There was no treatment-related death. Level 2 did not meet the criteria for the MTD. The relative dose intensities of the first five cycles were 91% for both 5-FU and irinotecan at level 1 and 86% for 5-FU and 93% for irinotecan at level 2. The response rates were 58% for all patients, and 69% for patients at level 2. CONCLUSION. Our results confirm that the modified Saltz regimen is safe and efficacious for Japanese patients. The recommended doses for phase II studies are irinotecan 100 mg/m2, 5-FU 500 mg/m2, and l-LV 10 mg/m2. PMID- 15549586 TI - Clinicopathological features of recurrence after radical surgery for nonmetastatic renal cell carcinoma. AB - BACKGROUND: The objective of this study was to clarify the clinicopathological features of recurrent renal cell carcinoma (RCC) in patients who had undergone curative surgical resection of primary disease. METHODS: The study included 171 patients treated by radical surgery for nonmetastatic RCC in our institution. Several clinicopathological factors were analyzed to determine differences between patients with and without disease recurrence. We further investigated predictive factors for disease recurrence, as well as prognostic factors after disease recurrence, using univariate and multivariate analyses. RESULTS: Thirty four of the 171 patients (19.9%) developed recurrence, and the median time from surgery to recurrence was 11 months (range, 1 to 146 months). The incidence of disease recurrence was significantly associated with sex, mode of detection, tumor size, pathological stage, tumor grade, and microvascular invasion (MVI). Univariate analysis showed that five factors--mode of detection, tumor size, pathological stage, tumor grade, and MVI--were found to be significant risk factors for recurrence after surgery, while only the pathological stage was an independent predictor of recurrence by multivariate analysis. After disease recurrence, the 1-, 3-, and 5-year cancer-specific survival rates were 66.3%, 45.9%, and 13.4%, respectively. Despite the absence of independent prognostic predictors after recurrence, recurrence more than 1 year after surgery and complete resection of recurrent lesions were shown to be significant prognostic factors by univariate analysis. CONCLUSION: These findings suggest that careful follow-up should be performed for patients showing a high pathological stage after radical surgery for nonmetastatic RCC, because of the higher probability of disease recurrence, and that after recurrence, intensive treatment should be considered, particularly for patients with unresectable recurrent disease occurring within 1 year after initial radical surgery. PMID- 15549587 TI - Noninvasive detection of alterations in chromosome numbers in urinary bladder cancer cells, using fluorescence in situ hybridization. AB - BACKGROUND: Bladder cancers have a high potential for recurrence and sometimes become invasive even in superficial cases. In this process, gene mutations in tumor suppressor genes such as p53, on chromosome 17, or p16, on chromosome 9, are thought to be important. In order to investigate whether the detection of alterations in chromosome number might be used as an alternative to invasive techniques for the assessment of clinical bladder cancer, fluorescence in situ hybridization (FISH) was employed to analyze chromosome numbers in a series of patients. METHODS: A total of 40 patients with transitional cell carcinomas (stages Ta to T4, including carcinoma in situ [CIS]) were examined for abnormal numbers of chromosomes 9 and 17, by FISH, using 41 and 42 samples of voided urine, respectively. Tumor grades were as follows: G1:G2:G3 = 4:21:17. Urinary cytology and presence of bladder tumor antigen (BTA) were also checked in the same samples. One hundred cells were examined in each sample, and abnormality was concluded to be present when more than 20% of cells demonstrated polysomy (defined as three or more chromosomes). RESULTS: Seventeen of 41 samples (41.5%) were abnormal with regard to chromosome 9, and 17 of 42 (40.5%) were abnormal for chromosome 17. Both chromosomes were affected in 13 cases, of which 8 were positive for urinary cytology and BTA. Univariate analysis, performed with urinary cytology, BTA, tumor grade, tumor stage, involvement of vessels, pattern of invasion, number of tumors, and prognosis as parameters, demonstrated a significant influence of urinary cytology (P = 0.0368 and P = 0.0278 for chromosomes 9 and 17, respectively), BTA (P = 0.0094 for chromosome 17), involvement of vessels (P = 0.0262 for chromosome 17), pattern of invasion (P = 0.0028 and P = 0.0327 for chromosomes 9 and 17), grade (P = 0.0213 and P = 0.0174 for chromosomes 9 and 17), and stage (P = 0.0457 for chromosome 17). All the other parameters also tended to be linked with the changes in chromosomes, except for tumor number. Multivariate analysis demonstrated significant differences for tumor grade (P = 0.0166) and pattern of invasion (P = 0.0006) for chromosome 9. CONCLUSION: Voided-urine FISH is an effective noninvasive method for the detection of altered chromosome numbers in bladder cancer cells, and may provide an indication of tumor progression when combined with urinary cytology and BTA. PMID- 15549588 TI - Doublet regimen of cisplatin plus docetaxel for second-line chemotherapy after prior therapy with cisplatin plus irinotecan for non-small cell lung cancer: a phase II study. AB - BACKGROUND: To evaluate the safety and efficacy of second-line chemotherapy with docetaxel and cisplatin for non-small cell lung cancer (NSCLC), we performed a phase II study. METHODS: The subjects were 25 patients with NSCLC, 75 years or younger, without organ dysfunction (performance status [PS], 0 to 2) in whom treatment with cisplatin and irinotecan had been ineffective or had been followed by recurrence or relapse. Four weeks or more after the end of the previous therapy, 60 mg/m2 of cisplatin and 60 mg/m2 of docetaxel were administered at intervals of 3 weeks. RESULTS: Observed toxicities of grade 3 or 4 included anemia (24% of patients), leukocytopenia (48%), neutropenia (76%), thrombocytopenia (4%), hepatic dysfunction (8%), and electrolyte abnormalities (4%). However, no severe nonhematologic adverse reactions occurred. The overall response rate was 32% (95% confidence interval, 13.7-50.3). The median time to disease progression was 98 days, and the median survival time was 257 days. CONCLUSION: Our results suggest that cisplatin and docetaxel can be used as second-line chemotherapy against NSCLC. But further, comparative, study of this combination should be performed in patients with good PS and organ function who have responded to prior platinum-based chemotherapy. PMID- 15549590 TI - Prognostic significance of thymidylate synthase in patients with metastatic colorectal cancer who receive protracted venous infusions of 5-fluorouracil. AB - BACKGROUND: This study was conducted to evaluate the prognostic significance of thymidylate synthase (TS) expression in the tumor tissue of patients with metastatic colorectal cancer (CRC) who received protracted venous infusions of 5 fluorouracil (5-FU). METHODS: We retrospectively analyzed the prognostic value of TS expression as compared with other clinical prognostic factors in 57 patients with metastatic CRC. RESULTS: On univariate analysis, survival was significantly related to TS expression (low vs high; P = 0.0015), alkaline phosphatase (ALP) level (<300 vs > or =300 IU/l; P = 0.0037), performance status (0 or 1 vs 2 or 3; P = 0.0073), and white blood cell count (<10,000/mm3 vs > or =10,000/mm3; P = 0.0001), with number of metastatic sites (1 vs > or =2; P = 0.06) approaching significance. On multivariate analysis, survival was significantly related to TS expression (hazard ratio [HR], 2.97) and ALP level (HR, 2.26). CONCLUSION: In patients with metastatic CRC who received protracted venous infusions of 5-FU, TS expression was related to survival independently of other established clinical prognostic factors. PMID- 15549589 TI - The efficacy of long-term oral chemotherapy with 5'-deoxy-5-fluorouridine and cyclophosphamide for recurrent breast cancer. AB - BACKGROUND: 5'-Deoxy-5-fluorouridine (5'-DFUR) is a prodrug of 5-fluorouracil (5 FU), which is known to be converted by thymidine phosphorylase (dThdPase). A recent preclinical study revealed that cyclophosphamide (CPA) upregulated dThdPase activity, specifically in tumor cells. The purpose of the present study was to examine the efficacy of long-term administration of 5'-DFUR/CPA for patients with recurrent breast cancer. METHODS: Fifteen breast cancer patients with recurrent tumors entered this study. Ten patients had bone metastasis, five had lung metastasis, and two had liver metastasis. Three patients had multiorgan metastases. All patients had had previous exposure to standard chemotherapy such as CAF (CPA, doxorubicin, and 5-FU) and CMF (CPA, methotrexate, and 5-FU). The patients were orally administered with daily doses of 5'-DFUR at 800-1200 mg and CPA at 200 mg for 2 weeks as induction therapy, followed by 2 weeks' rest (one to two cycles). Daily doses of 800 mg of 5'-DFUR and 100 mg of CPA (as maintenance therapy) were continuously administered thereafter. Ten of the 15 patients received the maintenance therapy alone. The treatment was continued for at least 24 months (average, 35.2 months). RESULTS: The main findings included a significant decrease in pain in nine patients with bone metastasis, and this effect continued for more than 2 years. As the pain decreased, the patients' quality of life (QOL) was improved. Liver metastasis was diminished in two out of two patients. Hematological toxicity of more than grade 3 was recognized in three patients, but only during the induction therapy. CONCLUSION: Oral administration of 5'-DFUR/CPA is well tolerated and useful for patients with recurrent breast cancer. PMID- 15549591 TI - Long-term survival in a patient with hepatocellular carcinoma with resection of a metastatic lymph node after percutaneous ethanol injection therapy. AB - Lymph node (LN) metastases during operation for hepatocellular carcinoma (HCC), but not during operations for other cancer, as many surgeons can attest. We performed partial hepatectomy with LN dissection in a man with LN metastasis from HCC, and long-term survival was achieved. In December 1993, at another hospital, a tumor, 4.2 cm in diameter, in this 73-year-old patient had been diagnosed as HCC. Transcatheter arterial embolization (TAE) was performed three times. Eighteen months after the operation, a swollen LN was discovered at the hepatic hilum and was treated by TAE once and by transcatheter arterial infusion (TAI) twice. However, the level of alpha-fetoprotein gradually increased and so percutaneous ethanol injection therapy (PEIT) was performed. Nevertheless, serum PIVKA-II (protein induced by vitamin K absence or antagonist II) continued to rise. The patient was referred to our hospital for further treatment. He underwent S5 subsegmentectomy with LN dissection. Histologically, the primary lesion consisted entirely of necrotic tissue. However, in the resected LN, there was residual cancer tissue near its periphery. We concluded that dissection of the affected LN offered the only chance for long-term survival, and that PEIT should be avoided for a metastatic lymph node. PMID- 15549592 TI - Esophageal cancer with colonic metastasis successfully treated by chemoradiotherapy followed by chemotherapy with S-1 and cisplatin. AB - A 51-year-old man was hospitalized for evaluation of dysphagia and bloody stool. Gastrointestinal endoscopy showed esophageal cancer invading the gastric fundus. A metastatic lesion was demonstrated in the sigmoid colon. The patient agreed to have concurrent chemoradiotherapy for the primary lesion, followed by additional chemotherapy. The first course included 30 Gy of radiotherapy given over 3 weeks, together with daily oral administration of S-1 (80 mg/m2 per day) for 2 weeks, and a 24-h infusion of cisplatin (70 mg/m2) on day 8. After a second course of chemoradiotherapy, four additional courses of chemotherapy with S-1 and cisplatin were administered, at 4-week intervals. After the additional chemotherapy, gastroscopy and colonoscopy showed disappearance of both the primary and the metastatic lesions. One year after his initial hospitalization, no recurrence of either the primary or the metastatic tumor lesions is evident. PMID- 15549593 TI - Complete response to irinotecan hydrochloride and nedaplatin in a patient with advanced ovarian clear cell carcinoma. AB - A 55-year-old multiparous woman was diagnosed with stage IIIc ovarian clear cell carcinoma. Three years after the first surgery and adjuvant chemotherapy with irinotecan hydrochloride and mitomycin C, she developed common iliac lymph node recurrence. Two cycles of chemotherapy with irinotecan hydrochloride and nedaplatin led to a complete response. Surgical resection revealed pathological complete response. The chemosensitivity of ovarian clear cell carcinoma has been reported to be very poor. No standard chemotherapeutic regimens for this carcinoma have been established. The present study is the first report of a pathological complete response in a patient with advanced ovarian clear cell carcinoma. Future studies are needed to confirm the efficacy of this regimen for this carcinoma. PMID- 15549594 TI - Acute gefitinib-induced pneumonitis. AB - A 60-year-old woman with non-small cell lung cancer was treated with gefitinib and developed acute pneumonitis on the third day. Pulmonary damage and/or interstitial lung disease (ILD) related to gefitinib was diagnosed clinically and radiographically. Despite the immediate withdrawal of gefitinib and the administration of high-dose steroid, the patient did not fully recover and finally died of respiratory failure on day 22 after gefitinib had commenced. This case cautions us that careful monitoring provides no guarantee of safeguarding patients against ILD caused by gefitinib. We should make the decision to treat patients with gefitinib very carefully until we can elucidate which patients are at high risk and which patients are likely to have a response to this drug. PMID- 15549595 TI - Malignant melanoma of pleura in a patient with giant congenital "bathing suit" hairy nevus. AB - An unusual case of malignant melanoma of the pleura in a patient with "bathing suit" type of giant congenital hairy nevus is presented. The treatment advocated and the outcome are discussed. PMID- 15549596 TI - Influence of multivalent counterions adsorption on Langmuir films. AB - It has been shown recently by neutron and X-ray reflectivity that nanometer-sized multivalent counterions (Keggin salts) can assemble as a dense monomolecular sublayer beneath a charged Langmuir monolayer of opposite sign. We have conducted experiments that examine the surface pressure isotherms of docosamine surfactant monolayers under such conditions and have shown that they undergo dramatic modifications when the Keggin salts are added. We model these experimental results by a close-packed sublayer of counterions on which charged surfactants can organize and form complexes. We then provide a thermodynamic description of the surface/sublayer system by giving an expression for surface free energy and surface pressure. We compare the results of this discrete model to traditional mean-field descriptions where the counterions form a diffuse continuous layer. The new features are: i) modifications in the shape of the surface pressure isotherm; ii) appearance of a phase separation in the surfactant layer. Finally, we show that the model is in satisfactory agreement with the experimental isotherms and a best fit yields numerical estimates of the different interaction parameters. PMID- 15549597 TI - Viscoelastic phase separation in polymer blends. AB - In this paper, the dynamics and morphology of viscoelastic phase separation in polymer blends is investigated based on the two-fluid model in two dimensions. At critical composition, we have carefully checked the role of shear modulus, without taking account of bulk modulus. The results show that the higher shear modulus component tends to form a dispersed phase in the intermediate stage of phase separation, if the difference between the shear moduli of the components is large enough. This is opposite to the role of bulk modulus, that the higher bulk modulus component forms a networklike pattern without taking account of the shear modulus even if it is the minority phase. The morphological formation is determined by the competition of opposite effects of shear modulus and bulk modulus. For polymer blends at critical composition, the bulk modulus difference leads to a networklike pattern formed by the higher modulus component in the intermediate stage of phase separation. But if the difference between the shear moduli of the components is large enough, a co-continuous structure is observed, resulting from the competition between shear and bulk moduli. For off-critical composition, difference in bulk modulus also leads to a networklike pattern of the component with higher bulk modulus in the intermediate stage of phase separation, but phase inversion is observed rapidly. A small difference between the shear moduli of the components can support the networklike pattern to continue for longer time. But the networklike pattern does not occur for large difference between shear moduli. PMID- 15549598 TI - pH-dependent multiple morphologies of novel aggregates of carboxyl-terminated polymide in water. AB - Rigid low-molecular-weight polyimide with two carboxyl ends (CPI) formed pH dependent multiple morphologies including dimpled-beads, porous aggregates and vesicles via self-assembly in aqueous media. This is a rapid and conventional approach to a series of pH-dependent morphologies of mono-component aggregates based on a readily attainable rigid homopolymer. PMID- 15549599 TI - Unusual behaviour of poly(ethylene-oxide) in aqueous mixtures. AB - The model system of poly(ethylene-oxide) or PEO, where the changing hydrogen-bond connectivity of the water has large effect on the conformation of the polymer chain, in mixtures of water and acetonitrile, is experimentally studied. The results show the existence of a threshold water content in the system at which the 3d connectivity of the water network begins. Unusual expansion of the polymer chain, an effect larger than that observed in either of the pure solvents, is seen. Upon addition of small amounts of a monovalent salt, binding of ion to polymer takes place in pure acetonitrile solutions. Salt ions begin to co ordinate with water molecules at the same solvent ratio as the threshold for water network formation. Ions now no longer complex to PEO; instead, hydrogen bonding of water to the polymer strongly dictates conformation in this regime. PMID- 15549600 TI - New investigation on the tetragonal liquid-crystalline phase or SmQ. AB - Chiral liquid crystals, for which twist and smectic order strongly compete, can exhibit complex architectures. This is the case of the SmQ phase which shows a high degree of 3D order (tetragonal or hexagonal) as well as a liquid signature. We have combined X-ray scattering and optical observations on a new compound exhibiting a SmQ phase to discriminate between the two models proposed by A.M. Levelut et al. (J. Phys. II 7, 981 (1997)) at the molecular level for the phase with the I4(1)22 space group. PMID- 15549601 TI - Development of an optical trap for microparticle clouds in dilute gases. AB - Long-duration experiments with clouds of microparticles are planned for the ICAPS facility on board the International Space Station ISS. The scientific objectives of such experiments are widespread and are ranging from the simulation of aerosol behaviour in Earth's atmosphere to the formation of planets in the early solar system. It is, however, even under microgravity conditions, impossible to sustain a cloud of free-floating, microscopic particles for an extended period of time, due to thermal diffusion and due to unavoidable external accelerations. Therefore, a trap for dust clouds is required which prevents the diffusion of the particles, which provides a source of relative velocities between the dust grains and which can also concentrate the dust to higher number densities that are otherwise not achievable. We are planning to use the photophoretic effect for such a particle trap. First short-duration microgravity experiments on the photophoretic motion of microscopic particles show that such an optical particle cloud trap is feasible. First tests of a two-dimensional trap were performed in the Bremen drop tower. PMID- 15549602 TI - Using diagnosis-related groups. The situation in the United Kingdom National Health Service and in Germany. PMID- 15549603 TI - Data envelopment analysis based bonus payments. Theory and application to inpatient care in the German state of Saxony-Anhalt. AB - This contribution proposes a reimbursement scheme combining yardstick competition and data envelopment analysis (DEA). The scheme reimburses incurred cost while offering a bonus for resource savings relative to a DEA-based cost norm, providing firms with an incentive to minimize production cost. The individual yardsticks are as close as possible to the actual cost levels, minimizing informational rents of firms. The proposed reimbursement scheme is applied to the hospital sector in Saxony-Anhalt based on data collected in 1999. The results indicate low regulation costs together with potentially large cost reductions due to the incentive structure of the reimbursement scheme. PMID- 15549604 TI - Exponential health utility. A characterization and comments on the contribution by Happich and Muhlbacher. AB - In a recent contribution Happich and Muhlbacher proposed an axiom of constant absolute trade-off of life-years for health states, and studied the family of QALY models satisfying this axiom under expected utility and mutual utility independence between life-years and health states. Here we provide a complete characterization of the family of QALY models satisfying these conditions. This family should not be mistaken for the family of multiplicative exponential QALY models; in particular, it violates the zero condition. PMID- 15549605 TI - CNF and DNT. AB - The actin cytoskeleton of mammalian cells is involved in many processes that affect the growth and colonization of bacteria, such as migration of immune cells, phagocytosis by macrophages, secretion of cytokines, maintenance of epithelial barrier functions and others. With respect to these functions, it is not surprising that many bacterial protein toxins, which are important virulence factors and causative agents of human and/or animal diseases, target the actin cytoskeleton of the host. Some toxins target actin directly, such as the C2 toxin produced by Clostridium botulinum. Moreover, bacterial toxins target the cytoskeleton indirectly by modifying actin regulators such as the low-molecular mass guanosine triphosphate (GTP)-binding proteins of the Rho family. Remarkably, toxins affect these GTPases in a bidirectional manner. Some toxins inhibit and some activate the GTPases. Here we review the Rho-activating toxins CNF1 and CNF2 (cytotoxic necrotizing factors) from Escherichia coli, the Yersinia CNF(Y) and the dermonecrotic toxin (DNT) from Bordetella species. We describe and compare their uptake into mammalian cells, mode of action, structure-function relationship, substrate specificity and role in diseases. PMID- 15549606 TI - Anthrax toxins. AB - Bacillus anthracis, the etiological agent of anthrax, secretes three polypeptides that assemble into toxic complexes on the cell surfaces of the host it infects. One of these polypeptides, protective antigen (PA), binds to the integrin-like domains of ubiquitously expressed membrane proteins of mammalian cells. PA is then cleaved by membrane endoproteases of the furin family. Cleaved PA molecules assemble into heptamers, which can then associate with the two other secreted polypeptides: edema factor (EF) and/or lethal factor (LF). The heptamers of PA are relocalized to lipid rafts where they are quickly endocytosed and routed to an acidic compartment. The low pH triggers a conformational change in the heptamers, resulting in the formation of cation-specific channels and the translocation of EF/LF. EF is a calcium- and calmodulin-dependent adenylate cyclase that dramatically raises the intracellular concentration of cyclic adenosine monophosphate (cAMP). LF is a zinc-dependent endoprotease that cleaves the amino terminus of mitogen-activated protein kinase kinases (Meks). Cleaved Meks cannot bind to their substrates and have reduced kinase activity, resulting in alterations of the signaling pathways they govern. The structures of PA, PA heptamer, EF, and LF have been solved and much is now known about the molecular details of the intoxication mechanism. The in vivo action of the toxins, on the other hand, is still poorly understood and hotly debated. A better understanding of the toxins will help in the design of much-needed anti-toxin drugs and the development of new toxin-based medical applications. PMID- 15549607 TI - The Helicobacter pylori vacuolating cytotoxin: from cellular vacuolation to immunosuppressive activities. AB - Helicobacter pylori is a highly successful bacterial pathogen of humans, infecting the stomach of more than half of the world's population. The H. pylori infection results in chronic gastritis, eventually followed by peptic ulceration and, more rarely, gastric cancer. H. pylori has developed a unique set of virulence factors, actively supporting its survival in the special ecological niche of the human stomach. Vacuolating cytotoxin (VacA) and cytotoxin-associated antigen A (CagA) are two major bacterial virulence factors involved in host cell modulation. VacA, so far mainly regarded as a cytotoxin of the gastric epithelial cell layer, now turns out to be a potent immunomodulatory toxin, targeting the adapted immune system. Thus, in addition to the well-known vacuolating activity, VacA has been reported to induce apoptosis in epithelial cells, to affect B lymphocyte antigen presentation, to inhibit the activation and proliferation of T lymphocytes, and to modulate the T cell-mediated cytokine response. PMID- 15549608 TI - Optimization of Serratia marcescens lipase production for enantioselective hydrolysis of 3-phenylglycidic acid ester. AB - Lipase production and cell growth of Serratia marcescens ECU1010 were optimized in shake flasks, with lipase production being enhanced 9.5-fold (4,780 U/l) compared with the initial activity (500 U/l). Optimal carbon and nitrogen sources were Tween-80 and peptone, and the optimal ratio of Tween-80 to peptone was 1:3. The optimized cultivation conditions were 25 degrees C and pH 6.5. Lipase activity, particularly specific activity, could be improved by decreasing the cultivation temperature from 35 to 25 degrees C. Enzyme stability was significantly improved by simple immobilization with synthetic adsorption resin no. 8244. After five reaction cycles, enzyme activity decreased only very slightly, while enantioselectivity of the preparation remained constant, and the ees (enantiomeric excess of the remaining substrate) achieved in all cases was higher than 97%. The resin-8244-lipase preparation can be used for efficient enantioselective hydrolysis of trans-3-(4'-methoxyphenyl)glycidic acid methyl ester [(+/-)-MPGM], a key intermediate in the synthesis of Diltiazem. PMID- 15549609 TI - Evidence for the existence of PAH-quinone reductase and catechol-O methyltransferase in Mycobacterium vanbaalenii PYR-1. AB - Polycyclic aromatic hydrocarbon (PAH) quinone reductase (PQR) and catechol-O methyltransferase (COMT), from the PAH-degrading Mycobacterium vanbaalenii PYR-1, were demonstrated to be constitutive enzymes located in the soluble fraction of cell extracts. PQR activities for the reduction of 9,10-phenanthrenequinone and 4,5-pyrene- quinone were 1.40+/-0.13 and 0.12+/-0.01 micromol min(-1) mg-protein( 1), respectively. The exogenous catechols alizarin, anthrarobin, 2,3 dihydroxynaphthalene and esculetin inhibited PQR activity. Anthrarobin (100 microM) and esculetin (100 microM) inhibited 4,5-pyrenequinone reduction by 64 92%. COMT was involved in the O-methylation of 1,2-dihydroxyphenanthrene to form 1-methoxy-2-hydroxyphenanthrene and 1,2-dimethoxyphenanthrene. Both pyrene and 1 hydroxypyrene were metabolized by M. vanbaalenii PYR-1 to form 1-methoxypyrene, 1 methoxy-2-hydroxypyrene, 1-hydroxy-2-methoxypyrene and 1,2-dimethoxypyrene. Among the catechols tested, anthrarobin showed the highest COMT activity (1.06+/-0.04 nmol/30 min(-1) mg-protein(-1)). These results suggest that the PQR and COMT activities of M. vanbaalenii PYR-1 may play an important role in the detoxification of PAH catechols. PMID- 15549610 TI - Spatial selectivity to manipulate portable objects in wedge-capped capuchins (Cebus olivaceus). AB - We studied the manipulative activity of five wedge-capped capuchins (Cebus olivaceus) confronted with different types of unfamiliar and portable objects: wooden blocks, plastic rings, spoons, and coconuts. Combinatorial manipulations involving two portable objects of the same type were quite frequent. The lately introduced objects, whatever their kind, appeared as the most attractive. Nevertheless, some objects remained very attractive throughout the overall experiment, especially the wooden blocks which elicited more combinatorial and striking behaviors than the other objects. Concerning space, we observed that the individuals choose specific locations to perform their manipulative acts. The spatial distributions of these acts were more concentrated, and less concordant between individuals, in the present study than in two others conducted with the same group but involving the manipulation of familiar objects. This suggests that individual differences were more marked when the subjects manipulated unfamiliar objects than when they manipulated familiar ones. This finding may have applications when the members of a group have to benefit from an enrichment of their environment. PMID- 15549611 TI - Expression of angiogenesis-related factors in lungs of patients with congenital diaphragmatic hernia and pulmonary hypoplasia of other causes. AB - Congenital diaphragmatic hernia (CDH) is a congenital disorder, complicated by pulmonary hypoplasia (PH) and pulmonary hypertension. Hypoplastic lungs have fewer and smaller airspaces than normal, with thicker interalveolar septa; the adventitia and media of pulmonary arteries are thickened, and the total size of the pulmonary vascular bed is decreased compared to normal. Although histological abnormalities in PH have been described, less is known about the underlying molecular mechanisms. Therefore, we have investigated a series of proteins, known to be involved in angiogenesis, including von Hippel-Lindau protein (pVHL), hypoxia-inducible factor-1a (HIF-1a), vascular endothelial growth factor (VEGF), fetal liver kinase 1 (Flk-1), and endothelial and inducible nitric oxide synthase (eNOS, iNOS) by immunohistochemistry on paraffin-embedded lung tissue of CDH patients ( n = 13), patients with lung hypoplasia due to other causes ( n = 20), and normal controls ( n = 33). pVHL was expressed more frequently in the arterial smooth muscle cells of CDH lungs compared with both other groups. Furthermore, HIF-1a was expressed less frequently in the endothelium of arteries, veins, and capillaries of CDH lungs as compared with both other groups. No differences were observed in the expression patterns of VEGF, Flk-1, eNOS, and iNOS between the different groups. Our data suggest a role for pVHL and HIF-1a in normal and abnormal pulmonary angiogenesis. The differential expression of these proteins may provide a molecular basis for the histological differences observed in the lung vessels of patients with CDH. PMID- 15549612 TI - Persistent right valve of the sinus venosus presenting as a right atrial cystic mass. PMID- 15549613 TI - "Myocardial infarction" in adolescents: do we have the correct diagnosis? AB - The evaluation of adolescents with chest pain, elevated cardiac enzymes, and abnormal electrocardiograms (ECGs) continues to pose diagnostic and management dilemmas. Myocardial infarction is an uncommon finding in this population and alternative diagnoses must be considered. Our database was retrospectively reviewed for adolescents age 16-18 years without prior cardiac history who underwent cardiac catheterization. Patients who presented with chest pain, elevated cardiac enzymes, normal ejection fraction, and abnormal ECGs were included. Management, diagnostic testing, and final diagnosis were reviewed. Nine adolescents (eight males and one female) without prior cardiac history were identified. The ECG findings in all patients were consistent with myocardial ischemia in a coronary distribution. Thrombotic coronary occlusion was not found in any patient. In adolescents without prior cardiac history of risk factors for myocardial infarction such as Kawasaki disease, familial hypercholesterolemia, or drug use who present with chest pain, multiple diagnoses must be considered even in the presence of focal ischemic ECG changes and elevated cardiac enzymes. Thrombolytic therapy or anticoagulation should be withheld until a definitive diagnosis of myocardial infarction has been made. Magnetic resonance imaging is the most useful tool to differentiate focal myocarditis from myocardial infarction. PMID- 15549614 TI - Hoarseness as the initial clinical presentation of anomalous left coronary artery from the pulmonary artery. AB - Anomalous origin of the left coronary artery from the pulmonary artery is a rare congenital anomaly that, if left untreated, will most often result in severe myocardial ischemia and significant morbidity and mortality. We report an unusual presentation of this defect in a 2-month-old infant who had an initial complaint of a "hoarse cry." We theorize that impingement of the recurrent laryngeal nerve due to dilatation of the pulmonary artery was the most likely etiology of the patient's symptoms. This case serves as an important reminder that serious congenital heart disease may present with any number of complaints and unusual findings. PMID- 15549615 TI - Severe coronary artery disease in the absence of supravalvular stenosis in a patient with Williams syndrome. AB - Williams syndrome is a complex syndrome comprising developmental abnormalities, craniofacial dysmorphic features, and cardiac anomalies. The most common cardiac anomaly is supravalvular aortic stenosis. We report a case of a 6-year-old girl with Williams syndrome who presented with decompensated heart failure due to ischemic cardiomyopathy. Her only significant cardiac anomaly was severe stenosis of the left main coronary artery. She subsequently died despite surgical revascularization. Isolated coronary anomalies are rare in Williams syndrome but should be considered especially in the presence of heart failure or ischemia. PMID- 15549618 TI - Coronary artery dilatation and aortic outflow tract enlargement in children with unicommissural aortic valves. AB - We evaluated the aortic outflow tract (AOT) and coronary artery dimensions in pediatric patients with unicommissural aortic valves. A retrospective review of an echocardiographic database identified 37 patients with unicommissural aortic valves. A total of 115 echocardiograms were reviewed, and the right coronary artery (RCA), left main coronary artery (LM), left anterior descending coronary artery aortic valve annulus, aortic root, sinotubular junction (STJ), and ascending aorta were measured and z scores determined. The aortic stenosis peak gradient and the amount of aortic regurgitation (AR) were also measured. The RCA diameter (z score, 1.85 +/- 1.8, p = 0.03) and LM diameter (z score, 1.74 +/- 1.47, p = 0.04) are significantly dilated, as are all the AOT measurements: aortic annulus (2.02 +/- 1.9, p = 0.02), aortic root (2.25 +/- 1.9, p = 0.02), STJ (2.22 +/- 1.74, p = 0.01), and ascending aorta (4.38 +/- 2.03, p < 0.001). Longitudinal follow-up showed that there was no significant variation over time in any variable. The AOT measurements were significantly correlated with each other. A trend was found in which an increasing amount of AR gave an increase in AOT measurements. The aortic gradient was not significantly associated with any measurement. Our study population demonstrated significant dilatation of the RCA and LM as well as the AOT. The dilatation of the AOT structures is likely caused by the same mechanism that accounts for the AOT dilatation in patients with bicommissural aortic valves. Dilatation of the coronary arteries may represent an intrinsic abnormality in the vessel wall. Further studies are needed to define possible changes. PMID- 15549617 TI - Use of covered stents in the management of coarctation of the aorta. AB - We report our experience with the use of covered stents for the management of coarctation of the aorta. From December 2001 to March 2004, nine patients (seven males; median age, 31 years; mean weight, 65 +/- 15 kg) underwent implantation. Indications included critical or atretic native coarctation (n = 4), patients >50 years of age (n = 2), associated patent ductus arteriosus (n = 1) or adjacent aneurysm (n = 1), and the presence of a circumferential fracture within a previously implanted stent (n = 1). The covered balloon-expandable Cheatham Platinum stent and the self-expandable stent graft Braile were employed. Adequate implantation was observed in all patients. Gradients were reduced from 54 +/- 14 to 3 +/- 8 mmHg and the coarctation site increased from 2.4 +/- 2.9 to 15.9 +/- 4.3 mm. The patent ductus arteriosus was immediately closed and the aneurysm excluded. Two patients >35 years with aneurysmal ascending aorta and metallic aortic prosthesis had aneurysm formation at follow-up, with one undergoing aneurysm exclusion using a Braile stent. Although covered stents are useful in the management of selected patients with coarctation, aneurysm formation may still occur in patients with markers of aortic wall weakness. Refinements in the deployment technique and/or the stent design are needed to eliminate this risk. PMID- 15549616 TI - Fungal endocarditis in neonates and children. AB - Fungal endocarditis in the neonate and pediatric population is associated with a high degree of morbidity and mortality, with diagnosis in the majority of cases made postmortem. The aim of this article is to review published literature on fungal endocarditis in children and to discuss the incidence due to a wide range of fungi, primarily Aspergillus and Candida species, as well as its diagnosis, including the importance of molecular-based approaches and various treatment regimes. PMID- 15549619 TI - Left ventricular noncompaction cardiomyopathy in association with trisomy 13. AB - In recent years, left ventricular noncompaction (LVNC) has been recognized as a distinct form of cardiomyopathy with its own clinical presentation and natural history. More than 100 cases of LVNC have been described in children. Although LVNC has been described in association with metabolic disorders such as Fabry's disease or genetic disorders such as Roifman's syndrome, this case represents the first report of LVNC in a child with trisomy 13. PMID- 15549620 TI - Aortic valvar atresia, interrupted aortic arch, and quadricuspid pulmonary valve: a rare combination. AB - Aortic atresia and interrupted aortic arch is a rare cardiac combination. Review of the literature revealed nine cases. We present two patients with this combination and the additional finding of quadricuspid pulmonary valves, one of which was severely stenotic. In the latter patient, an aortopulmonary window was present. The other had a unique blood supply to the brachiocephalic arteries and ascending aorta from systemic collateral arteries. To the best of our knowledge, the association of a quadricuspid pulmonary valve with this combination has not been previously reported. PMID- 15549621 TI - Vascular tumors of the heart in infants and children: case series and review of the literature. AB - Primary tumors of the heart are rare in children, of which vascular tumors comprise a small subgroup. We present the clinical, histopathologic, and imaging findings in six children with vascular tumors of the heart and review the findings of 36 previously published cases. We observed three intramuscular hemangiomas of the small-vessel type in older children, two congenital hemangiomas in infants, and one malignant polymorphous hemangioendothelioma. Intramuscular hemangiomas did not respond to corticosteroid and were biologically distinct from the congenital hemangiomas, both of which exhibited regression with pharmacotherapy. Age at diagnosis appears to predict histologic type, tumor location, and clinical presentation. PMID- 15549622 TI - Quantifying and minimizing radiation exposure during pediatric cardiac catheterization. AB - This study reports findings from evaluations of new technologies to measure radiation exposure during pediatric cardiac catheterization procedures. A strategy of pulsed fluoroscopy and low power settings resulted in significantly lower patient radiation exposure compared to conventional 60 frames/sec, high power settings during fluoroscopy. During radiofrequency ablation procedures, thyroid and thoracic skin sites outside the direct fluoroscopic field received minimal radiation exposure. Intrathoracic radiation exposure was measured with the use of an esophageal dosimeter. In conclusion, strategies to reduce total radiation exposure should be employed, radiation dose should be measured, and assessment of radiation skin injury should be included in post-catheterization assessment. PMID- 15549623 TI - The pediatric cardiology pharmacopoeia: 2004 update. AB - Seven years ago, Pediatric Cardiology published the first version of a review article outlining the various medications used in the field of heart diseases in children. This article is an update and expansion to what we have previously presented. Therapeutic intervention, both surgical and through cardiac catheterization, has enabled cure and palliation of an increasingly expanding spectrum of diseases at earlier ages and with more complex lesions. Refinement of these procedures includes more advanced tools as well as the support of an expanding armament of pharmacopoeia used to stabilize and support patients before, during, and after such procedures. In addition to updating previously published data regarding inotropes, antiarrhythmics, vasodilators, diuretics, sedatives, and analgesics as well as a variety of miscellaneous medications, this article describes the use of pulmonary medications frequently needed in patients with congestive heart failure, pulmonary edema, and chronic lung disease. We also describe the difficult management of withdrawal as a result of use of sedatives and analgesics. The most recent recommendation for subacute bacterial endocarditis prophylactic antibiotic regimens is also described. PMID- 15549624 TI - A prospective randomized trial of closing laparoscopic trocar wounds by transcutaneous versus subcuticular suture or adhesive papertape. AB - BACKGROUND: Several methods for closure of trocar wounds are known in laparoscopic surgery. The choice of technique (mostly transcutaneous or subcuticular suture or adhesive papertape) is often based on the surgeon's personal experience. Thus, the objective of this trial was to assess the impact of these closure methods on potential complications of wound healing, cosmetic outcome, and patient satisfaction. METHODS: Sixty patients undergoing operative laparoscopic surgery for gynecologic indications were enrolled in this prospective randomized trial. Five-millimeter port-site incisions were closed either with subcuticular or transcutaneous absorbable sutures (4-0 polyglactin 910) or with adhesive papertape. Postoperative complications, pain, and patient satisfaction with scars were evaluated at 3-month follow-up after operation using a questionnaire. RESULTS: Data from 52 patients who returned the questionnaire were analyzed. Dissatisfying cosmetic results were reported significantly more frequently after subcuticular sutures (p < 0.05). Assessment of patient satisfaction with cosmetic outcome on a visual-analogue scale revealed significantly better results after transcutaneous skin closure than with other approaches (p < 0.05). Adverse wound healing (e.g., infections and dehiscence) was observed most frequently in the subcuticular suture group. Also, the rate of painful scars was highest with this technique. CONCLUSIONS: Transcutaneous closure with absorbable suture material seems to be the most suitable technique for closure of laparoscopic port-site incisions. PMID- 15549625 TI - Stapled laparoscopic cystgastrostomy: a series with 15 cases. AB - BACKGROUND: The goal of this study was to assess the clinical outcome of patients undergoing laparoscopic stapled cystgastrostomy for pancreatic pseudocysts in contact with the posterior wall of the stomach. METHODS: We performed a case note review of all patients who have undergone stapled laparoscopic cystgastrostomy in Norwich, UK. The cystgastrostomy was fashioned through an anterior gastrotomy using a vascular ETS stapling device in all cases. RESULTS: Fifteen patients have undergone stapled laparoscopic cystgastrostomy. The procedure was completed successfully in 12 patients. Three procedures were converted to open surgery for technical reasons. There were no complications due to bleeding from the cystgastrostomy. Early complications included systemic sepsis (one), bleeding gastric ulcer (one) and pseudocyst recurrence due to partial closure of the cystgastrostomy (two). No late recurrences or other complications have been found at a median follow-up of 37 months. CONCLUSION: Stapled laparoscopic cystgastrostomy is a safe and effective procedure for draining pancreatic pseudocysts in contact with the posterior wall of the stomach. The use of a hemostatic stapling device to fashion the cystgastrostomy may reduce the risk of catastrophic hemorrhage from the pseudocyst wall. PMID- 15549626 TI - Long-term results of laparoscopic vs open resections for rectal cancer in 124 unselected patients. PMID- 15549627 TI - A 6-year experience with the Swedish adjustable gastric band Prospective long term audit of laparoscopic gastric banding. AB - BACKGROUND: In morbid obesity conservative therapy often fails to reduce overweight permanently. As a consequence, several bariatric surgical procedures have been developed to achieve permanent excess weight loss. Among these, the laparoscopic restrictive procedures seem to be the least invasive. The aim of this prospective study was to assess and analyze the effects, complications, and outcomes after the implantation of the Swedish adjustable gastric band (SAGB) in long-term follow-up. METHODS: All consecutive patients with implantation of a SAGB between August 1996 and August 2002 were prospectively investigated. The placement of the SAGB was done by laparoscopy in all cases. Success was rated by the reduction of body mass index (BMI) excess weight loss (EWL), and reduction of comorbidities. "Nonresponders" to SAGB were defined as <30% EWL after a 3-year follow-up. Band-related complications were recorded and classified. Patient's outcome was assessed after 6 months and subsequently each year postoperatively. RESULTS: A total of 190 patients received a SAGB, 97% of whom could be followed up with a mean follow-up period of 39.4 months (duration of follow-up, 6-72). During follow-up, a significant reduction or improvement of BMI, EWL, and comorbidities were found. Nineteen percent of patients were identified as nonresponders. Early intraoperative and postoperative complications related to SAGB were one perforation of the gastric fundus (0.5%), one conversion (0.5%), one bleeding (0.5%), and two band infections (1.1%). The SAGB-related complications encountered during long-term follow-up were three port problems (1.6%), four band migrations (2.1%), five slipping/pouch dilatations (2.6%), and two band leakages (1.1%). All intra- and postoperative SAGB-related complications accounted for a total morbidity of 10.5%. Operative mortality was 0%. The overall reoperation rate was 8.5%. CONCLUSIONS: In long-term follow-up, SAGB is safe and effective. Our results demonstrate a significant EWL of 50% during the first 24 months. However, patient selection has to be improved to reduce the nonresponder rate. SAGB leads to a significant reduction of obesity-related comorbidities. SAGB is an attractive alternative in the surgical treatment of morbid obesity. PMID- 15549628 TI - Significant weight loss after laparoscopic Nissen fundoplication. AB - BACKGROUND: Laparoscopic Nissen fundoplication (LNF) has evolved as a gold standard in antireflux surgery. However, the association between body weight and gastroesophageal reflux disease (GERD) is still unclear, and no data are available concerning the effect of fundoplication on body weight. We present the first report elucidating the impact of LNF on body weight in GERD patients with special emphasis on patients' quality of life. METHODS: From July 2000 to March 2003, LNF was carried out in 213 patients (85 women and 128 men) after thorough preoperative examination including clinical interview with standardized assessment of symptoms and quality of life (QoL), endosocopy, barium swallow, 24 h pH-metry, and manometry. Follow-up investigations were performed 3 and 12 months after LNF obtainable from 209 patients (98.1%) and 154 patients (72.3%), respectively. RESULTS: The mean body mass index (BMI) decreased significantly after LNF (27.6 +/- 5.6 kg/m(2) before LNF vs 26.0 +/- 3.8 kg/m(2) after LNF, p < 0.001). Twelve months after LNF, neither a tendency toward a renewed increase nor a further decrease in BMI was observable. The average body weight loss was 3.9 kg. BMI reduction was higher in women than in men (p < 0.002), and obese patients lost more weight than lean patients (p < 0.001). There was no association between BMI reduction and dysphagia. Plasma cholesterol and triglyceride levels did not change after LNF. The mean general score of the Gastrointestinal Quality of Life Index markedly improved (90.1 +/- 21.3 before LNF vs 118.0 +/- 16.2 after LNF, p < 0.01), as did the GERD-Health Related Quality of Life Index (21.9 +/- 6.4 before LNF vs 3.5 +/- 2.7 after LNF, p < 0.001). However, there was no association between changes in BMI and QoL. CONCLUSION: LNF leads to significant and persistent body weight loss. PMID- 15549629 TI - Advantages and limits of robot-assisted laparoscopic surgery: preliminary experience. AB - BACKGROUND: In the last few years, robotics has been applied in clinical practice for a variety of laparoscopic procedures. This study reports our preliminary experience using robotics in the field of general surgery to evaluate the advantages and limitations of robot-assisted laparoscopy. METHODS: Thirty-two consecutive patients were scheduled to undergo robot-assisted laparoscopic surgery in our units from March 2002 to July 2003. The indications were cholecystectomy, 20 patients; right adrenalectomy, two points; bilateral varicocelectomy, two points; Heller's cardiomyotomy, two points; Nissen's fundoplication, two points; total splenectomy, one point; right colectomy, one point; left colectomy, 1 point; and bilateral inguinal hernia repair, one point. In all cases, we used the da Vinci surgical system, with the surgeon at the robotic work station and an assistant by the operating table. RESULTS: Twenty nine of 32 procedures (90.6%) were completed robotically, whereas three were converted to laparoscopic surgery. Conversion to laparoscopy was due in two patients to minor bleeding that could not be managed robotically and to robot malfunction in the third patient. There were no deaths. Median hospital stay was 2.2 days (range, 2-8). CONCLUSIONS: The main advantages of robot-assisted laparoscopic surgery are the availability of three-dimensional vision and easier instrument manipulation than can be obtain with standard laparoscopy. The learning curve to master the robot was >or= 10 robotic procedures. The main limitations are the large diameter of the instruments (8 mm) and the limited number of robotic arms (maximum, three). We consider these technical shortcomings to be the cause for our conversions, because it is difficult to manage bleeding episodes with only two operating instruments. The benefit to the patient must be evaluated carefully and proven before this technology can become widely accepted in general surgery. PMID- 15549630 TI - Conversion rates in laparoscopic colorectal surgery: a predictive model with, 1253 patients. AB - BACKGROUND: This study aimed all develop a mathematical model for predicting the conversion rate for patients undergoing laparoscopic colorectal surgery. METHOD: This descriptive single-center study used routinely collected clinical data from 1,253 patients undergoing laparoscopic surgery between November 1991 and April 2003. A two-level hierarchical regression model was used to identify patient, surgeon, and procedure-related factors associated with conversion of laparoscopic to open surgery. The model was internally validated and tested using measures of discrimination and calibration. Exclusion criteria for laparoscopic colectomy included a body mass greater than 50, lesion diameter exceeding 15 cm, and multiple prior major laparotomies (exclusive of appendectomy, hysterectomy, and cholecystectomy). RESULTS: The average conversion rate for the study population was 10.0% (95% confidence interval [CI], 8.3-11.7%). The independent predictors of conversion of laparoscopic to open surgery were the body mass index (odds ratio [OR], 2.1 per 10 Americans Society of Anesthesiology units increase), (ASA) grade 3 or 4, 1 or 2 (OR, 3.2, 5.8), type of resection (low rectal, left colorectal, right colonic vs small/other bowel procedures; OR, 8.82, 4.76, 2.98), presence of intraoperative abscess (OR, 3.60) or fistula (OR, 4.73), and surgeon seniority (junior vs senior staff OR, 1.56). The model offered adequate discrimination (area under receiver operator characteristic curve, 0.74) and excellent agreement (p = 0.384) between observed and model-predicted conversion rates (range of calibration, 3-32% conversion rate). CONCLUSIONS: Laparoscopic conversion rates are dependent on a multitude of factors that require appropriate adjustment for case mix before comparisons are made between or within centers. The Cleveland Clinic Foundation (CCF) laparoscopic conversion rate model is a simple additive score that can be used in everyday practice to evaluate outcomes for laparoscopic colorectal surgery. PMID- 15549631 TI - Comparison of open and laparoscopic secure placement of peritoneal dialysis catheters. AB - BACKGROUND: A major and frustrating complication of peritoneal dialysis catheter placement is mechanical outflow obstruction, which may be caused by catheter tip migration. Therefore, a secure and correct positioning of the catheter is important to minimize this risk. This technique is easily accomplished by a laparoscopic approach. METHODS: The outcomes of 50 patients in whom peritoneal dialysis catheters were inserted laparoscopically with a secure catheter placement technique were compared with those of 52 patients who underwent an open surgical technique using a stiff wire as guidance for the catheter. The data were prospectively collected but not randomized. All the patients had virgin abdomens, and all the procedures were undertaken or supervised by one surgeon. RESULTS: Catheter migration occurred in six patients (12%) in the open group, as compared with none in the laparoscopic group (p = 0.027). There were no significant differences in catheter survival between the two groups. CONCLUSIONS: The laparoscopic technique with secure placement of the catheter lowered the incidence of catheter migration, but did not increase the catheter survival. PMID- 15549632 TI - Assessment of pulmonary function in patients before and after laparoscopic and open esophagogastric surgery. AB - BACKGROUND: Laparoscopy is a technique used in various surgical procedures. Few studies in the literature compare stress between laparoscopic and open surgery used for esophagogastric surgical procedures. Pulmonary function is known to be significantly affected in open surgeries, increasing postoperative morbidity and mortality. The current study aimed to assess pulmonary function in patients before and after open and laparoscopic esophagogastric surgery. METHODS: For this study, 75 patients were divided into two groups: 50 patients undergoing laparoscopy and 25 patients undergoing open surgery. The following parameters were determined by spirometry before and after surgery: forced expiratory volume in the first second (FEV(1)), forced vital capacity (FVC), and forced expiratory flow in the midexpiratory phase (FEF(25-75%)). RESULTS: A decrease in FEV(1,) FVC, and FEF((25-75%)) was observed in the two groups on postoperative days 2, 3, and 4, as compared with the preoperative period. Likewise, FEV(1) and FVC showed a significant reduction on postoperative days 2, 3, and 4 in the patients who underwent to open surgery, but only on the day 2 in those who underwent to laparoscopic surgery. A significant decrease in FEF((25-75%)) was observed only on postoperative day 2 in the group that underwent open surgery. Significant differences in FEV(1) between the groups were observed on postoperative days 2, 3, and 4. No significant difference in FVC was noted between the groups, and a difference in FEF((25-75%)) was observed only on postoperative day 4. CONCLUSIONS: Postoperative pulmonary dysfunction was more important for the patients undergoing open surgery than for those undergoing laparoscopic surgery. PMID- 15549633 TI - Open vs thorascopic surgical management of bronchogenic cysts. AB - BACKGROUND: The aim of this study was to compare the operative outcome in children undergoing open vs thoracoscopic resection of bronchogenic cysts. METHODS: The medical records of children who underwent the resection of bronchogenic cysts from 1990 through 2000 were reviewed. Four cyst resections were performed by the open technique and five using a thoracoscopic procedure. The age of the patients, length of hospital stay, duration of drainage, operating time, and outcome were investigated. RESULTS: The mean age of patients undergoing the open procedure was 3 years and 3 months; the mean age for thoracoscopy patients was 7 years and 10 months (p < 0.05). The operating time for the open procedure was 70 +/- 25 min; for the laparoscopic procedure, it was 78 +/- 6 min (p, NS), except in one case with a main bronchial tail that required conversion (320 min). Duration of surgical drainage was 6.5 +/- 3 days for the open procedure and 2.5 +/- 1 days for the thoracoscopic one (p < 0.05). Hospital stay for open patients was 12 days +/- 0 days; it was 6 +/- 1.6 days for thoracoscopic patients (p < 0.01). There were no deaths. The thoracoscopic procedure failed once due to a main bronchial tail and had to be converted to an open procedure. Other early complications included a bronchopulmonary infection after an open cyst excision and an atelectasis after a thoracoscopic cyst excision. Late complications included one reoperation for incomplete excision in each of the two groups. CONCLUSION: Bronchogenic cyst resection can be performed safely. For complete treatment of these patients, total excision of the wall cyst is needed. In selected patients, the thoracoscopic procedure may decrease the duration of surgical drainage and length of hospital stay without increasing the operating time or MSK for complications. PMID- 15549634 TI - Results of thoracoscopic sympathectomy for the treatment of axillary and palmar hyperhidrosis with respect to compensatory hyperhidrosis and dry hands. AB - BACKGROUND: We examined the results of thoracoscopic sympathectomy (TS) for palmar and axillary hyperhidrosis with respect to operative method, symptom control, patient satisfaction and complications. METHODS: We performed a retrospective review of patient records with mail and telephone questionnaire follow-up of 55 patients (15 men) with a median age of 26 years (range, 15-52) who underwent TS between February 1994 and December 2001. RESULTS: There were no differences in complication rates between those having bilateral TS (n = 23) and those having unilateral procedures (n = 20) with a median follow-up of 21 months (range, 2-94). Forty-three patients returned questionnaires (response rate, 78%). Forty patients (93%) were satisfied with the results. Thirty-four patients (79%) noted compensatory hyperhidrosis and 22 (51%) excessively dry hands. CONCLUSION: Despite high rates of compensatory sweating, the majority of patients are very satisfied with the results. The high rate of excessively dry hands is a previously unreported finding and important to discuss when obtaining consent. PMID- 15549635 TI - First inventory of resorption lacunae on rods and plates of trabecular bone as observed by scanning electron microscopy. AB - In the present study a novel systematic distribution scheme of resorption lacunae (RL) was applied using scanning electron microscopy. RL, classified as either reticulate patch resorption lacunae (RPR) or as longitudinally extended resorption lacunae (LER) [11, were analyzed and quantified according to their localizations on rods (middle, nodes or both) and plates (central or peripheral) in standardized segments from the femoral head of 24 Caucasian subjects without bone disease. Age and gender variations were explored. No clear gender-related distribution pattern could be detected on plates. On rods of males, however, the distribution of RL tended to be higher at the nodes, but seemed to be more prevalent in the middle or extended from the middle to the nodes of rods in females. Certain other non-conclusive tendencies in relation to age, gender, type of RL and localization were observed. PMID- 15549636 TI - Localization of pigment epithelium-derived factor in growing mouse bone. AB - Pigment epithelium-derived factor (PEDF) is a potent anti-angiogenic factor found in a wide range of fetal and adult tissues, where it is thought to play a role in the regulation of angiogenesis during development. The temporal expression of PEDF during endochondral bone formation has not previously been reported. In this study, we analysed the expression pattern of PEDF in growing mouse hindlimbs from newborn day one through to maturation at week 9, using immunohistochemistry and in situ hybridization. PEDF expression was demonstrated in chondrocytes within the resting, proliferative and upper hypertrophic zones of the epiphyseal growth plate. The pattern of expression was consistent throughout the developmental stages of the mouse. In addition, PEDF was expressed by osteoblasts lining the bone spicules in the ossification zone of metaphyseal bone, as well as by osteoblasts lining cortical periosteum. These novel results demonstrate that PEDF is developmentally expressed in both cartilage and bone cells during endochondral bone formation, and strongly suggest that it may play a regulatory role in the processes of chondrocyte and osteoblast differentiation, endochondral ossification, and bone remodelling during growth and development of long bones. PMID- 15549638 TI - Modulation of collagen fibrillogenesis by dentinal proteoglycans. AB - Studies have identified different pools of proteoglycan (PG) species present within the unmineralized matrix of the predentine, the transitional phase at the predentine-dentine interface and the mineralized dentine. These PGs alter with respect to the chemical nature of their glycosaminoglycan (GAG) chains and as a result of extracellular processing of the macromolecule in the matrix. The present study has examined the influence of the PGs isolated from these phases and the influence of the attached GAG chains, upon their ability to control collagen fibrillogenesis. PGs isolated from the three phases were characterized and determined to contain a mixture of decorin and biglycan. Results have indicated that predentine PGs, which are substituted with a higher proportion of dermatan sulfate, significantly delayed fibril formation while ultimately promoting the formation of thicker fibrils. Removal of the GAG chains further delayed fibrillogenesis, leading to the formation of thinner fibrils, compared with the collagen-only control. PGs isolated from predentine-dentine, which contained a higher proportion of chondroitin sulfate, also significantly delayed fibrillogenesis, resulting in thicker collagen fibrils. GAG chains attached to the predentine-dentine interface PGs played a role in the timing of fibrillogenesis with fibril formation initiated at the same time as the collagen control, but yielding thicker fibrils. Dentine PGs significantly inhibited fibrillogenesis and fibril thickness over concentrations of 50-25 microg/mL protein. In conclusion, the PGs isolated from the distinct phases have indicated differing roles in the orchestrated organization of the extracellular matrix during dentinogenesis, with roles for both the core protein and attached GAG chains indicated. PMID- 15549637 TI - Ascorbic acid deficiency, iron overload and alcohol abuse underlie the severe osteoporosis in black African patients with hip fractures--a bone histomorphometric study. AB - Osteoporosis and femoral neck fractures (FNF) are uncommon in black Africans although osteoporosis accompanying iron overload (from traditional beer brewed in iron containers) associated with ascorbic acid deficiency (oxidative catabolism by iron) has been described from sub-Saharan Africa. This study describes histomorphometric findings of iliac crest bone biopsies and serum biochemical markers of iron overload and of alcohol abuse and ascorbic acid levels in 50 black patients with FNFs (29 M, 21 F), age 62 years (40-95) years (median [min max]), and in age- and gender-matched black controls. We found evidence of iron overload in 88% of patients and elevated markers of alcohol abuse in 72%. Significant correlations between markers of iron overload and of alcohol abuse reflect a close association between the two toxins. Patients had higher levels of iron markers, i.e., siderin deposits in bone marrow (P < 0.0001), chemical non heme bone iron (P = 0.012), and serum ferritin (P = 0.017) than controls did. Leukocyte ascorbic acid levels were lower (P = 0.0008) than in controls. The alcohol marker mean red blood cell volume was elevated (P = 0.002) but not liver enzymes or uric acid. Bone volume, trabecular thickness, and trabecular number were lower, and trabecular separation was greater in patients than in controls, all at P < 0.0005; volume, surface, and thickness of osteoid were lower and eroded surface was greater, all at P < 0.0001. There was no osteomalacia. Ascorbic acid deficiency accounted significantly for decrease in bone volume and trabecular number, and increase in trabecular separation, osteoid surface, and eroded surface; iron overload accounted for a reduction in mineral apposition rate. Alcohol markers correlated negatively with osteoblast surface and positively with eroded surface. Relative to reported data in white FNF patients, the osteoporosis was more severe, showed lower osteoid variables and greater eroded surface; FNFs occurred 12 years earlier and were more common among men. We conclude that the osteoporosis underlying FNFs in black Africans is severe, with marked uncoupling of resorption and formation in favor of resorption. All three factors--ascorbic acid deficiency, iron overload, and alcohol abuse--contributed to the osteoporosis, in that order. PMID- 15549639 TI - Soy isoflavones may protect against orchidectomy-induced bone loss in aged male rats. AB - Evidence from several studies suggests that soy protein and/or its isoflavones may have beneficial effects on bone in postmenopausal women and animal models who have osteoporosis. The present study examined the dose-dependent effects of soy isoflavones in the context of soy protein or casein on the male skeleton. Thirteen-month-old male Fisher 344 rats were orchidectomized (ORX; 5 groups) or sham-operated (Sham; 1 group) and immediately placed on dietary treatments for 180 days. Diets were semi-purified and the protein source was either casein (Sham and ORX; controls), casein with two added doses of isoflavones (Iso1; 600 mg/kg diet and Iso2; 1200 mg/kg diet), soy protein with normal isoflavones content (Soy; 600 mg/kg diet), or soy protein with added isoflavones (Soy+; 1200 mg/kg diet). A 7% loss of whole body bone mineral density (BMD) was observed due to orchidectomy; however, the ORX induced BMD loss was significantly reduced to 4.3 and 4.7 % with the Soy and Soy+, respectively. Both doses of isoflavones in conjunction with casein also reduced the loss of whole body BMD, albeit not significantly different from ORX control animals. Trabecular bone histomorphometric analysis of the proximal tibia further supported the bone sparing role of soy isoflavones as indicated by higher percent bone volume and trabecular number, and lower trabecular separation. We conclude that isoflavones exert modest beneficial effects on the male skeleton whether provided with casein or a soy protein. PMID- 15549640 TI - Ovariectomy reduces the gelatinolytic activity and expression of matrix metalloproteinases and collagen in rat molar extraction wounds. AB - Osteoporosis is commonly associated with estrogen deficiency. However, the mechanisms by which the lack of this hormone causes bone loss are poorly understood. The bone structure of the oral cavity seems to be affected by estrogen deficiency, since a delayed healing process after tooth extraction has been observed after ovariectomy in rats. The aim of this study was to describe the effect of the absence of estrogen on the expression and activity of matrix metalloproteinases (MMC)-2 and -9 and expression of types I and III collagens in the alveolar granulation tissue of young female rats after tooth extraction. Sixty-six, four-week-old female rats underwent bilateral ovariectomies (OVX) or sham operations. Three weeks later, both first and second mandibular molars were extracted and the animals were killed by cervical dislocation 3, 5, or 7 days after tooth extraction. The granulation tissues were collected from the extracted alveolar sockets and used for zymographic, Western blot, or reverse transcription polymerase chain reaction (RT-PCR) analysis. There was a gradual increase on the expression of all studied proteins as well as MMP-2 and -9 activities in the periods after surgery. In contrast, OVX animals showed a significant decrease in the gelatinolytic activities and expression of MMP-2 and -9 and types I and III collagens. The results presented here in suggest that the absence of estrogen may possibly contribute to the delayed alveolar wound healing by interfering with the extracellular matrix turnover. PMID- 15549641 TI - Maternal history of osteoporosis and femur geometry. AB - Most studies that have examined the role of skeletal factors in the relationship between an individual's family history of fracture or osteoporosis and their fracture risk have focused on bone density. In this study, we expanded the scope of skeletal factors to include geometric properties (subperiosteal width, section modulus, cortical thickness, and buckling ratio) in addition to areal bone mineral density (BMD). We compared these skeletal factors at the femur neck and shaft by self-reported maternal history of osteoporosis (OP HX) from 5334 non Hispanic whites, ages > or =20 years in the Third National Health and Nutrition Examination Survey (NHANES III, 1988-94). A total of 213 men and 315 women reported a positive OP HX (e.g., their biological mother had sustained a hip fracture after age 50 years or had a physician's diagnosis of osteoporosis). Differences in bone density and geometry by OP HX were examined after adjusting for potential confounding variables. Several bone parameters differed significantly by OP HX in both sexes at the femur neck, but none differed at the femur shaft. At the neck, those with a positive OP HX had values that differed by approximately 3% to 4% (lower for BMD, bone mineral content (BMC), cross sectional area, and cortical thickness; higher for buckling ratios) from those with a negative OP HX (P < 0.05). The magnitude of these relationships was similar in both sexes, but differences were greater in younger versus older adults. In conclusion, both men and women with a positive maternal history of osteoporosis may be at greater risk of femur neck fracture owing to thinner cortices and lower BMC, which in turn results in potentially greater cortical instability (buckling ratio) at this skeletal site. PMID- 15549642 TI - Soft drink and milk consumption, physical activity, bone mass, and upper limb fractures in children: a population-based case-control study. AB - Carbonated beverages have been reported to increase fracture risk in children but the mechanism is unclear. The aim of this population-based case-control study was to investigate the association between soft drink and milk consumption, physical activity, bone mass, and upper limb fractures in children aged 9-16 years. A total of 206 fracture cases and 206 randomly selected individually matched controls were studied. There were 47 hand fractures; 128 wrist and forearm fractures, and 31 upper arm fractures. An interviewer-administered questionnaire was utilized to retrospectively assess last-year physical activity (including television, computer, and video watching) and to recall the average weekly consumption of milk, colas, and total carbonated drinks. Bone mass at the spine, hip, and total body was assessed by dual-energy X-ray absorptiometry (DXA) and metacarpal morphometry. For total fractures, none of the above drink types was significantly different between cases and controls. For wrist and forearm fractures, there was a positive association between cola drink consumption and fracture risk (OR 1.39/unit, 95% CI: 1.01, 1.91). Cola consumption was significantly correlated with television, computer, and video watching (r = 0.20, P = 0.001) but not bone mineral density or milk drinks. After adjustment for television, computer, and video watching and bone mineral density, the association between cola drinks and fracture risk became nonsignificant (OR 1.31/unit, 95% CI: 0.94, 1.83). No association with other fracture sites was observed. In conclusion, cola, but not total carbonated beverage consumption, is associated with increased wrist and forearm fracture risk in children. However, this association is not independent of other factors and appears to be mediated by television watching and bone mineral density but not by decreased milk intake. PMID- 15549643 TI - Bone mineral density, carotid artery intimal medial thickness, and the vitamin D receptor BsmI polymorphism in Mexican American women. AB - Low bone mineral density (BMD) is a predictor of cardiovascular mortality, suggesting that osteoporosis and cardiovascular disease may share common risk factors. We assessed the relationship between BMD and intimal medial thickening (IMT) of the common carotid artery, a marker of sub-clinical atherosclerosis, in 471 women examined as part of the San Antonio Family Osteoporosis Study, a population-based study of osteoporosis risk conducted in Mexican American families. Because of the documented role of vitamin D metabolism in bone metabolism and its possible role in cardiovascular function, we further evaluated whether allelic variation at the vitamin D receptor locus (VDR) influenced joint variation in BMD and IMT. The association of BMD with IMT depended on age, with low BMD being correlated with high IMT in older women, but with low IMT in younger women [age by IMT interaction effects significant at the spine (P = 0.042), radius ultradistal (P = 0.010), and hip (P = 0.006)]. In all women, the VDR BsmI BB genotype was associated with significantly higher forearm BMD (P = 0.005 for both radius ultradistal and midpoint), higher IMT (P = 0.05), and higher spine BMD in older women (P = 0.06), but not with hip BMD. The association of the VDR genotype with IMT was independent of its association with BMD. Although a functional consequence of the BsmI polymorphism on vitamin D metabolism has not been established, these findings support a possible biological relationship among VDR, bone metabolism, and atherosclerosis. We conclude that VDR polymorphisms may be one of multiple factors influencing the joint risk of atherosclerosis and osteoporosis. PMID- 15549644 TI - APOE haplotypes influence bone mineral density in Caucasian males but not females. AB - Low bone mineral density (BMD) is one of the most important risk factors for osteoporosis. Apolipoprotein E (APOE) has been considered as a candidate gene for osteoporosis because of its influence on osteoblast uptake of lipoprotein-borne vitamin K. Using the quantitative transmission disequilibrium test QTDT, we examined linkage and/or association of APOE and BMD at the lumbar spine and the total hip in a sample of 387 Caucasian nuclear families with 715 parents and 953 children. The children were aged 20-50 years and female offspring were premenopausal as well. Four single nucleotide polymorphisms (SNP1-4) in the APOE gene, 4-locus haplotypes and 2-locus haplotypes (epsilon1, epsilon2, epsilon3, epsilon4 isoforms, reconstructed by SNP3 and SNP4) were analyzed. In the whole sample and the female offspring families we found no evidence of linkage or association for either single SNP or haplotype with BMD at the two studied skeletal sites. In the male offspring families, within-family associations were observed at the haplotypes CGTC (P = 0.001), GGTT (P = 0.002), and GATC (P = 0.006) for the lumbar spine BMD, and GATC (P = 0.008) for the total hip BMD. These data suggested that in our studied Caucasian population, APOE may have effects on BMD variation in males but not females. Further studies with a larger sample size are required to confirm such results. PMID- 15549645 TI - Heritability of spinal trabecular volumetric bone mineral density measured by QCT in the Diabetes Heart Study. AB - The heritability of trabecular volumetric bone mineral density (BMD) determined by quantitative computed tomography (QCT) has not yet been reported. The purpose of this study was to investigate the heritability of BMD as determined by QCT and DXA in 124 women and 120 men (age 39-83 years, BMI 17-75, 84% type 2 diabetics) from 101 families (232 sibling pairs) in the Diabetes Heart Study. Volumetric BMD had a heritability (h2) estimate of 0.73 (SE = 0.15, P < 0.0001) at the lumbar spine and 0.71 (SE = 0.15, P < 0.0001) at the thoracic spine. Areal BMD heritability estimates were 0.56 for PA spine, 0.43 for total hip, 0.43 for femoral neck, 0.45 for distal radius, 0.42 for mid-radius, and 0.52 for whole body (all P < 0.01). After accounting for familial correlation using generalized estimating equations, volumetric BMD was inversely associated with age (r = 0.52, P < 0.0001) and duration of diabetes (r = -0.24, P < 0.01) and positively associated with body weight (r = 0.25, P < 0.01). In multivariate analysis, adjustment for age, sex, and race lowered the h2 estimates for volumetric BMD at the lumbar (h2 = 0.41, P < 0.01) and thoracic (h2 = 0.48, P < 0.001) spine, increased the h2 estimate for areal BMD at the mid radius (h2 = 0.58, P < 0.0001), and had little effect on the h2 estimate for areal BMD at other sites (h2 = 0.41-0.55, all P < 0.01). Additional adjustment for BMI, duration of diabetes, and physical activity had little effect on the h2 estimates for volumetric BMD or areal BMD except at the hip where they were lowered (h2 = 0.31 0.33, all P < 0.05). These data suggest that, like areal BMD, volumetric BMD is highly heritable and may be used in designing linkage studies to locate genes governing bone metabolism. PMID- 15549646 TI - Simultaneous exposure of excess fluoride and calcium deficiency alters VDR, CaR, and calbindin D 9 k mRNA levels in rat duodenal mucosa. AB - Fluoride ingestion reduces intestinal calcium absorption; its molecular basis has not been studied. We studied the mRNA expression of calcium-sensing receptor (CaR), vitamin D receptor (VDR) and calbindin D 9 k (D 9 k) by northern blot analysis in the duodenal mucosa of rats. Weanling pups fed with chow diet containing adequate calcium (0.5% w/w) and drinking water (NaF < 1 ppm) served as controls (Group I) and were studied at 9 and 15 weeks. The pups, born to rats fed with a calcium-deficient diet (0.03%) and excess fluoride water (NaF 50 ppm), were continued on the same diet and water (Group II) until 9 weeks of age. Subsequently, Group II rats were divided into 4 subgroups; 3 subgroups with fluoride free water [II-A adequate calcium, II-B excess calcium (Ca 2%) and II-D calcium deficient], whereas II-C received fluorinated water and adequate calcium diet until 15 weeks. At 9 weeks, as compared to group-I, group-II had decreased VDR (P < 0.001) and D 9 k mRNA (P < 0.001), whereas CaR mRNA levels increased (P < 0.05). At 15 weeks, as compared to group-I, VDR mRNA further reduced in group II-D (P < 0.001) and II-C (P < 0.001), whereas it increased in group II-A. Removal of fluoride ingestion and calcium replenishment increased D 9 k mRNA expression, maximally in adequate calcium group (P < 0.001), while it was further reduced in group II-C (P < 0.001). CaR expression decreased significantly in all the groups. We conclude that excess fluoride reduces the mRNA levels of VDR and D 9 k in the duodenal mucosa of rats, thereby possibly reducing calcium absorption. Calcium supplementation with simultaneous fluoride removal improves their expression. PMID- 15549647 TI - Effect of the proportion of organic material in bone on thermal decomposition of bone mineral: an investigation of a variety of bones from different species using thermogravimetric analysis coupled to mass spectrometry, high-temperature X-ray diffraction, and Fourier transform infrared spectroscopy. AB - Thermogravimetric analysis linked to mass spectrometry (TGA-MS) shows changes in mass and identifies gases evolved when a material is heated. Heating to 600 degrees C enabled samples of bone to be classified as having a high (cod clythrum, deer antler, and whale periotic fin bone) or a low (porpoise ear bone, whale tympanic bulla, and whale ear bone) proportion of organic material. At higher temperatures, the mineral phase of the bone decomposed. High temperature X ray diffraction (HTXRD) showed that the main solids produced by decomposition of mineral (in air or argon at 800 degrees C to 1000 degrees C) were beta-tricalcium phosphate (TCP) and hydroxyapatite (HAP), in deer antler, and CaO and HAP, in whale tympanic bulla. In carbon dioxide, the decomposition was retarded, indicating that the changes observed in air and argon were a result of the loss of carbonate ions from the mineral. Fourier transform infrared (FTIR) spectroscopy of bones heated to different temperatures, showed that loss of carbon dioxide (as a result of decomposition of carbonate ions) was accompanied by the appearance of hydroxide ions. These results can be explained if the structure of bone mineral is represented by [Formula: see text] where V(Ca) and V(OH) correspond to vacancies on the calcium and hydroxide sites, respectively, and 2-x-y = 0.4. This general formula is consistent in describing both mature bone mineral (i.e., whale bone), with a high Ca/P molar ratio, lower HPO4(2-) content, and higher CO3(2-) content, and immature bone mineral (i.e., deer antler), with a low Ca/P ratio, higher HPO4(2-), and lower CO3(2-) content. PMID- 15549648 TI - Enhanced marrow adipogenesis and bone resorption in estrogen-deprived rats treated with the PPARgamma agonist BRL49653 (rosiglitazone). AB - Thiazolidinediones are insulin-sensitizing agents and in clinical use for the treatment of type II diabetes. Under specific experimental conditions, these molecules induce adipogenic differentiation of mesenchymal precursor cells at the expense of osteoblasts in vitro, suggesting possible negative effects on the skeleton. We measured effects of the thiazolidinedione BRL49653 on bone tissue of intact and estrogen-deprived skeletally mature adult female Wistar rats (6-9 months old). Weight gain and decreased plasma triglyceride levels confirmed the effectiveness of the treatment. However, no change in bone mass or fat marrow volume was observed in intact rats treated for 8 weeks with 5, 10, or 20 mg/kg of BRL49653. Study of marrow cultures established at necropsy revealed a higher responsiveness to adipogenic differentiation protocols of cultures established from the 10-mg/kg group compared to vehicle control. In a second study, the effects of thiazolidinedione treatment on the skeleton of estrogen-deprived rats were investigated. Application of 10 mg/kg of BRL49653 for 12 weeks resulted in enhanced bone loss (+31%; pQCT) and increased fat marrow volume (+117%; histomorphometry) compared to vehicle-treated OVX control. Interestingly, osteoblast number was comparable in both cases. Bone resorption parameters were significantly increased in the treatment group (+27% osteoclast number, +30% eroded surface). Enhanced bone loss due to treatment was consistently observed in the tibia, femur, and the lumbar spine. Our data indicate that thiazolidinediones may enhance bone loss induced by estrogen deprivation. PMID- 15549649 TI - Osteoclastogenesis inhibitory factor/osteoprotegerin reduced bone loss induced by mechanical unloading. AB - Skeletal unloading resulting from space flight and prolonged immobilization causes bone loss. Such bone loss ostensibly results from a rapid increase in bone resorption and subsequent sustained reduction in bone formation, but this mechanism remains unclear. Osteoclastogenesis inhibitory factor/osteoprotegerin (OCIF/OPG) is a recently identified potent inhibitor of osteoclast formation. We studied effects of OPG administration on tail-suspended growing rats to explore the therapeutic potential of OPG in the treatment and prevention of bone loss during mechanical unloading, such as that which occurs during space flight. Treatment with OPG in tail suspension increased the total bone mineral content (BMC g) of the tibia and femur and the total bone mineral density (BMD g/cm2) of the tibia. Moreover, treatment with OPG prevented reduction not only of BMC and BMD, but also of bone strength occurring through femoral diaphysis. Treatment with OPG in tail-suspended rats improved BMC, BMD and bone strength to levels of normally loaded rats treated with vehicle. Treatment with OPG in normally loaded rats significantly decreased urinary excretion of deoxypyridinoline, but the effect of OPG in tail suspension was unclear. These results indicate that OPG may be useful in inhibiting bone loss-engendered mechanical unloading. PMID- 15549651 TI - A model of navigation-induced currents in inland waterways and implications for juvenile fish displacement. AB - The likely extension of commercial inland navigation in the future could increase hazards directly impacting on the nurseries of freshwater fish, especially for smaller individuals with limited swimming abilities. One limitation of the evaluation of inland navigation on fish assemblages is the lack of suitable hydraulic models. This article presents a hydraulic model to assess the increase of navigation-induced physical forces due to higher vessel speed, length, and drought in a low-flowing waterway related to maximum swimming performance of fish to (1) foresee hazards of enhancement of inland navigation, (2) derive construction measures to minimize the hydraulic impact on small fish, and (3) improve fish recruitment in waterways. The derived model computed current velocities induced by passing commercial vessels in inland waterways experimentally verified and parameterized in a German lowland waterway. Results were linked with a model of maximum fish swimming performance to elucidate consequences for freshwater fish populations. The absolute magnitude of navigation-induced current limits the availability of littoral habitats for small fish. Typical navigation-induced current velocities of 0.7-1 m/s in the straight reaches of waterways will be maintained by fish longer than 42 mm only. Smaller juveniles unable to withstand those currents could become washed out, injured, or displaced. In contrast, in small local bays, the navigation-induced current declined significantly. According to our model, in a 20-m extended bay, the return current drops below 0.11 m/s, corresponding to the maximum swimming speed of a 9-mm-long fish. Thus, enhancing shoreline development by connecting oxbows, tributaries, and especially by purpose-built bays limits the impact on fish recruitment without restricting navigation resulting in more precautionary and sustainable inland navigation. PMID- 15549652 TI - Genomic structure and functional analysis of promoter region of somatolactin gene of sea bream (Sparus aurata). AB - Somatolactin (SL) is a pituitary hormone belonging to the growth hormone prolactin family and is produced in the intermediate lobe of teleosts. The SL gene was isolated from a sea bream genomic library and found to be composed of 5 exons distributed within a 9-kb length of DNA. Sequence analysis of the proximal promoter region showed the presence of a classical TATA box located 59 bp upstream from the initial start ATG codon, 5 consensus sequences corresponding to the Pit-1 binding element, and a putative CREB site. In CHO cells cotransfected with the DNA from 2 plasmids, one encoding sea bream Pit-1 under Rous sarcoma virus long terminal repeat regulation and one encoding the SL promoter driving the expression of luciferase, Pit-1 was found to enhance the expression of luciferase. Only one Pit-1 binding site was necessary for enhancement. Analysis by immunoblots of in vitro culture of pituitaries of Sparus aurata showed that several agents, including estradiol, verapamil, and phorbol myristate acetate, had different inhibitory effects on SL and growth hormone released to the culture medium. PMID- 15549653 TI - Molecular cloning and functional characterization of fatty acyl desaturase and elongase cDNAs involved in the production of eicosapentaenoic and docosahexaenoic acids from alpha-linolenic acid in Atlantic salmon (Salmo salar). AB - Fish are the only major dietary source for humans of omega-3 highly unsaturated fatty acids (HUFAs) and with declining fisheries farmed fish such as Atlantic salmon (Salmo salar) constitute an increasing proportion of the fish in the human diet. However, the current high use of fish oils, derived from wild capture marine fisheries, in aquaculture feeds is not sustainable in the longer term and will constrain continuing growth of aquaculture activities. Greater understanding of how fish metabolize and biosynthesize HUFA may lead to more sustainable aquaculture diets. The study described here contributes to an effort to determine the molecular genetics of the HUFA biosynthetic pathway in salmon, with the overall aim being to determine mechanisms for optimizing the use of vegetable oils in Atlantic salmon culture. In this paper we describe the cloning and functional characterization of 2 genes from salmon involved in the biosynthesis of HUFA. A salmon desaturase complementary DNA, SalDes, was isolated that include an open reading frame of 1362 bp specifying a protein of 454 amino acids. The protein sequence includes all the characteristics of microsomal fatty acid desaturases, including 3 histidine boxes, 2 transmembrane regions, and an N terminal cytochrome b(5) domain containing a heme-binding motif similar to that of other fatty acid desaturases. Functional expression in the yeast Saccharomyces cerevisiae showed SalDes is predominantly an omega-3 delta5 desaturase, a key enzyme in the synthesis of eicosapentaenoic acid (20:5n-3) from alpha-linolenic acid (18:3n-3). The desaturase showed only low levels of delta6 activity toward C(18) polyunsaturated fatty acids. In addition, a fatty acid elongase cDNA, SalElo, was isolated that included an open reading frame of 888 bp, specifying a protein of 295 amino acids. The protein sequence of SalElo included characteristics of microsomal fatty acid elongases, including a histidine box and a transmembrane region. Upon expression in yeast SalElo showed broad substrate specificity for polyunsaturated fatty acids with a range of chain lengths, with the rank order being C(18) > C(20) > C(22). Thus this one polypeptide product displays all fatty acid elongase activities required for the biosynthesis of docosahexaenoic acid (22:6n-3) from 18:3n-3. PMID- 15549655 TI - Imperatorin inhibits T-cell proliferation by targeting the transcription factor NFAT. AB - In our ongoing research into anti-inflammatory compounds from medicinal plants in the Mediterranean area, we have isolated several furanocoumarins from the roots of Oppopanax chironium (L.), and have evaluated them for activity related to T cell functionality. Heraclenin (1) and imperatorin (2) significantly inhibited T cell receptor-mediated proliferation in human primary T cells in a concentration dependent manner. In transient transfection experiments with a plasmid containing the IL-2 promoter we found that imperatorin is a potent inhibitor of IL-2 gene transcription. To further characterize the inhibitory mechanisms of imperatorin at the transcriptional level, we examined the DNA-binding and transcriptional activities of NF-kappaB, NFAT, and AP-1 transcription factors in Jurkat T cells. We found that imperatorin inhibited both the NFAT binding to DNA and transcriptional activities, without affecting significantly the activation of the NF-kappaB and AP-1 transcription factors. These findings provide new insights into the molecular mechanisms involved in the immunomodulatory and anti inflammatory activities of natural furanocoumarins. PMID- 15549656 TI - Casuarinin protects cultured MDCK cells from hydrogen peroxide-induced oxidative stress and DNA oxidative damage. AB - Casuarinin has been shown to be an antioxidant in acellular experiments. This study was designed to assess the ability of casuarinin, extracted from Terminalia arjuna, to protect cultured Madin-Darby canine kidney (MDCK) cells against H2O2 mediated oxidative stress. A comparison with trolox, a hydrosoluble vitamin E analogue was performed. MDCK cells were pretreated with casuarinin or trolox for 1 h, then exposed to H2O2. After incubation with 0.8 mM H2O2 for 1 h, casuarinin caused a decrease in intracellular peroxide production as shown by dichlorofluorescein (DCF) fluorescence in a concentration-dependent manner. After 3 h exposure to 8 mM H2O2, the percentage of intracellular glutathione (GSH) negative cells was reduced in the casuarinin-treated group. Addition of 32mM H2O2 to MDCK cells for 3 h induced an increase in the percentage of cells containing 8 oxoguanine but the level of such cells declined in casuarinin-treated cells. These results show that casuarinin is more effective against H2O2-induced oxidative damage than trolox. The data suggest that casuarinin attenuates H2O2 induced oxidative stress, decreases DNA oxidative damage and prevents the depletion of intracellular GSH in MDCK cells. PMID- 15549657 TI - Anti-inflammatory and antinociceptive effects of sinapyl alcohol and its glucoside syringin. AB - In the present study, syringin, isolated by activity-guided fractionation of the ethyl acetate (EtOAc) extracts of the stem bark of Magnolia sieboldii, and sinapyl alcohol, the hydrolysate of syringin, were evaluated for anti inflammatory and antinociceptive activities. Sinapyl alcohol (20, 30 mg/kg/day, p. o.) inhibited increased vascular permeability by acetic acid in mice and reduced acute paw edema by carrageenan in rats more so than syringin. When analgesic activity was measured using the acetic acid-induced writhing test and the hot plate test, sinapyl alcohol was much more potent than syringin in a mouse model. In addition, sinapyl alcohol more potently inhibited lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor (TNF)-alpha production by macrophages than syringin. Consistent with these observations, the expression levels of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 was reduced by sinapyl alcohol in a concentration dependent manner. These results suggest that the anti-inflammatory and antinociceptive effects of syringin after oral administration may be attributed to its in vivo transformation to sinapyl alcohol. PMID- 15549658 TI - Enhancement of antitumor effects of paclitaxel (taxol) in combination with red ginseng acidic polysaccharide (RGAP). AB - We have recently reported that red ginseng acidic polysaccharide (RGAP), isolated from Korean red ginseng (Panax ginseng C.A. Meyer), shows immunomodulatory and antitumor activities, mainly mediated by the nitric oxide (NO) production of macrophages. This compound may be used in cancer therapy alone or in combination with other chemotherapeutic agents. The synergistic effect of RGAP and paclitaxel (taxol) was evaluated to develop new biological response modifiers in cancer therapy. The present study demonstrates a synergistic antitumor effect of RGAP and paclitaxel in mice transplanted with sarcoma 180 and B16 melanoma. Combined treatment with paclitaxel (5 or 15 mg/kg) and RGAP (25 mg/kg) resulted in a 28.6 or 42.8 % increase in the life span of ICR mice bearing sarcoma 180 tumor cells, while no obvious effect was seen on sole paclitaxel treatment. When a combination of paclitaxel (10 mg/kg) and RGAP (100 mg/kg) was administered to C57BL/6 mice implanted with B16 melanoma, the tumor weight per mouse also decreased by 76.3 %, suggesting that RGAP may be used as an adjuvant in medicinal applications of paclitaxel. The augmented antitumor effect of paclitaxel is supposed to be the result of the immunomodulating antitumor effect of RGAP. RGAP, having B cell specific mitogenic activity, induced the secretion of interleukin-6 (IL-6) in spleen cells in a concentration-dependent manner (5 to 500 microg/microL). RGAP also restored the proliferation of splenocytes and NK cell activity suppressed by paclitaxel. Flow cytometric analysis of splenocytes in mice treated with paclitaxel showed a significant increase of CD11b+ cells. Additionally, a synergistic effect of RGAP and paclitaxel was found to effect an increased tumoricidal activity of macrophages. The above results suggest that clinical trials of RGAP as an adjuvant in cancer chemotherapy of paclitaxel are highly feasible. PMID- 15549659 TI - A study of the cardioprotective effect of breviscapine during hypoxia of cardiomyocytes. AB - Breviscapine is a flavonoid extracted from Erigeron breviscapus. Hand.-Mazz, and it has been reported that breviscapine can activate K+ channels and block Ca2+ channels. In this paper, we studied the cardioprotective effects of breviscapine on electrocardiogram (ECG) changes (ST-segment elevation), infarction size in dog heart subjected to myocardial infarction caused by left coronary artery ligation and lactate dehydrogenase (LDH) leakage, changes of intracellular free Ca2+ levels, apoptosis and necrosis in cultured neonatal rat cardiomyocytes subjected to hypoxia. Additionally, the effect of breviscapine on myocardial oxygen consumption was detected in dog myocardium in vitro. The results showed that breviscapine treatment (1 mg/kg, 2 mg/kg and 4 mg/kg) significantly reduced ST segment elevation and infarction size in hearts subjected to myocardial infarction, that breviscapine treatment (14.29 microg/mL, 28.57 microg/mL and 57.14 microg/mL) significantly decreased oxygen consumption in myocardium, and that breviscapine treatment (5 microg/mL, 10 microg/mL and 20 microg/mL) significantly reduced LDH leakage, intracellular free Ca2+ levels, apoptosis and necrosis in cardiomyocytes subjected to hypoxia. In conclusion, the present study indicates that breviscapine is in favor of myocardial protection. PMID- 15549660 TI - Polyacetylenic compounds and butanol fraction from Bidens pilosa can modulate the differentiation of helper T cells and prevent autoimmune diabetes in non-obese diabetic mice. AB - Compelling evidence suggests that infiltrating CD4+ type I helper T (Th1) cells in the pancreatic islets play a pivotal role in the progression of diabetes in non-obese diabetic (NOD) mice. We demonstrate in the present report that a butanol fraction of B. pilosa suppressed the development of diabetes, helped maintain levels of blood sugar and insulin in NOD mice in a dose-dependent manner and elevated the serum IgE levels regulated by Th2 cytokines in NOD mice. Moreover, the butanol fraction inhibited the differentiation of naive helper T (Th0) cells into Th1 cells but enhanced their transition into type II helper T (Th2) cells using an in vitro T cell differentiation assay. Two polyacetylenic compounds, 2-beta-D-glucopyranosyloxy-1-hydroxy-5(E)-tridecene-7,9,11-triyne and 3-beta-D-glucopyranosyloxy-1-hydroxy-6(E)-tetradecene-8,10,12-triyne, identified from the butanol fraction also prevented the onset of diabetes like the butanol fraction. The latter compound showed a stronger activity for T cell differentiation than the former. In summary, the butanol fraction of B. pilosa and its polyacetylenes can prevent diabetes plausibly via suppressing the differentiation of Th0 cells into Th1 cells and promoting that of Th0 cells into Th2 cells. PMID- 15549661 TI - Evidence of the regulatory effect of Ginkgo biloba extract on skin blood flow and study of its effects on urinary metabolites in healthy humans. AB - Ginkgo biloba extract has been advocated for the improvement of blood circulation in circulatory disorders. This study investigated the effect of the Gingko biloba extract EGb 761 on skin blood flow in healthy volunteers and accompanying changes in urinary metabolites. Twenty-seven healthy middle-aged subjects participated in a randomized, double-blind, placebo-controlled, crossover study. Subjects received 240 mg/d EGb 761 or placebo for periods of 3 weeks. Skin blood flow was measured on the forefoot using laser Doppler flowmetry and changes in urinary metabolites were identified by a combination of nuclear magnetic resonance (NMR) spectroscopy and multivariate data analysis (MVDA). These measurements were performed on 24-h urine samples collected at the end of the intervention periods. Following EGb 761 treatment, overall mean skin blood flow was significantly reduced as compared with placebo. Remarkably, the change of skin blood flow after EGb 761 intervention was proportionally related to blood flow after placebo treatment: subjects showed either an increased, decreased or unaltered skin blood flow. NMR/MDVA analyses showed that urinary metabolic patterns differed depending on the change in baseline blood flow after treatment with EGb 761. The present findings substantiate that EGb 761 has a multi-directional modulating action on blood flow in healthy subjects and support findings of a vasoregulatory role of this extract. Moreover, the results indicate that metabolic fingerprinting provides a powerful means to identify biochemical markers that are associated with functional changes. PMID- 15549662 TI - Effect of 13-epi-sclareol on the bacterial respiratory chain. AB - 13-epi-sclareol is a labdane-type diterpene isolated from the resinous exudates of the medicinal plant species Pseudognaphalium cheiranthifolium (Lam.) Hilliard et Burtt. and P. heterotrichium (Phil.) A. Anderb. This compound has antibacterial activity only against Gram-positive bacteria, showing a bactericidal and lytic action. The interaction of 13- epi-sclareol with the bacterial respiratory chain was analyzed. The compound inhibited oxygen consumption of intact Gram-positive cells, but not with Gram-negative bacteria. The compound inhibited NADH oxidase and cytochrome c reductase activities, while coenzyme Q reductase and the cytochrome c oxidase activities were not affected. These results suggest that the target site of 13-epi-sclareol is located between coenzyme Q and cytochrome c. Using cytoplasmic membrane fractions, the results of the analysis of the enzyme activities associated with the respiratory chain complexes were the same for both Gram-positive and Gram-negative bacteria, indicating that the compound has no access to the cytoplasmic membrane of intact Gram-negative bacteria. Thus, the Gram-negative envelope may act as a physical barrier that prevents the access of this compound to the site of action. PMID- 15549663 TI - Daphnetoxin interacts with mitochondrial oxidative phosphorylation and induces membrane permeability transition in rat liver. AB - The effects of daphnetoxin on isolated rat liver mitochondria and freshly isolated rat hepatocytes were investigated. Daphnetoxin (in the microM range) increased mitochondrial state 4 respiration and decreased both state 3 and FCCP uncoupled respiration. The transmembrane potential was strongly depressed by daphnetoxin in a concentration-dependent manner. The protonophoric activity of daphnetoxin was evidenced by the induction of mitochondrial swelling in hyposmotic K+ acetate medium in the presence of valinomycin. In isolated hepatocytes, daphnetoxin decreases intracellular ATP and simultaneously increases ADP and AMP concentrations. The addition of uncoupling concentrations of daphnetoxin to Ca2+-loaded mitochondria treated with Ruthenium Red results in non specific membrane permeabilization, as evidenced by mitochondrial swelling in isosmotic sucrose medium. Mitochondrial swelling in the presence of Ca2+ was prevented by cyclosporine A and was drastically inhibited by catalase and dithiothreitol, indicating the participation of mitochondrial generated reactive oxygen species in this process. From this study we can conclude that the bioenergetic lesion promoted by daphnetoxin seems to be sufficient to explain the lethal hapatocyte injury. PMID- 15549664 TI - Pharmacokinetics of baicalin in rats and its interactions with cyclosporin A, quinidine and SKF-525A: a microdialysis study. AB - Baicalin, a flavone glucuronide derived mainly from the root of Scutellaria baicalensis, has been used in traditional Chinese medicine as an anti inflammatory and anti-viral agent. To explore whether the disposition of baicalin is related to multidrug resistance P-glycoprotein (P-gp), baicalin (3, 10 and 30 mg kg(-1); i. v.) was injected to rats for a pharmacokinetic study using microdialysis coupled with HPLC. The results indicate that baicalin goes through hepatobiliary excretion against a concentration gradient based on the blood-to bile distribution ratio (AUCbile/AUCblood), but that AUCblood or AUCbile did not show any dose-related increase in the range from 3 to 30 mg kg(-1). Coadministration of cyclosporin A (CsA) or quinidine (both are P-gp inhibitors) was used to delineate the role of P-gp on baicalin disposition, while SKF-525A (a cytochrome P450 inhibitor) could specifically inhibit the cytochrome P450 catalysis of baicalin without crossing with P-gp function. Both CsA and quinidine promoted the active transport of baicalin into bile and reduced its level in blood, and this result was the same as that obtained by treating with SKF-525A. Hence, the association of the involvement of P-gp in active baicalin efflux into bile seems to be excluded since CsA and quinidine are also cytochrome P450 inhibitors. In addition, baicalin was not detected in the brain striatum after treating with baicalin alone in the present study. Also, neither CsA nor quinidine co-administered with baicalin is able to induce measurable levels of baicalin in rat brain, which suggests that baicalin might not be able to pass through the blood-brain barrier (BBB). PMID- 15549665 TI - Steroids from the roots of Cynanchum stauntonii. AB - A chemical investigation of the roots of Cynanchum stauntonii has resulted in the characterization of a new hydroperoxide with a 13,14 : 14,15-disecopregnane-type skeleton, named stauntonine (1), together with three related compounds, anhydrohirundigenin (2), anhydrohirundigenin monothevetoside (3), and glaucogenin C mono- D-thevetoside (4). Their structures were established by spectroscopic methods, including X-ray crystallographic diffraction analysis of stauntonine that confirmed its relative stereochemistry. The compound 1 showed dose-dependent relaxation on aortic rings with endothelium contracted by phenylepherine or KCl. PMID- 15549666 TI - Phylogenetic analysis of the DNA sequence of the non-coding region of nuclear ribosomal DNA and chloroplast of Ephedra plants in China. AB - Twenty-four Ephedra plants belonging to 8 species grown in the northern and western parts of China were phylogenetically analyzed for their non-coding DNA sequences, internal transcribed spacers (ITSs) of nuclear ribosomal DNA as well as trnL intron and intergenic spacers between trnL and trnF (trnL/ trnF) of the chloroplast. Based on the ITS sequences, the 8 species could be divided into 3 groups: Group 1 (Ephedra intermedia, E. sinica, E. przewalskii), Group 2 (E. equisetina, E. monosperma, E. gerardiana), and Group 3 (E. likiangensis, E. minuta). The species classified into Group 1 grow mainly in the north, Group 3 in the south and Group 2 in the center, suggesting their genetic and geographic relationships. A specific primer set was designed to classify the 3 groups by routine PCR. Combined analysis of ITS and trnL/ trnF differentiated the 8 Ephedra species. PMID- 15549667 TI - Antiviral and antiplasmodial spirodihydrobenzofuran terpenes from the fungus Stachybotrys nephrospora. AB - Two known spirodihydrobenzofuran terpenes (1 and 2) were isolated from a mycelium extract of the fungus Stachybotrys nephrospora BCC 3900. Compound 1 (Mer-NF5003F or stachybotrydial) exhibited potent antiviral activity (the IC50 value of 4.32 microg/mL) comparable to the standard drug, acyclovir, while compound 2 was inactive against the HSV-1 virus. Both 1 and 2 possessed antiplasmodial activity (IC50 values of 0.85 and 0.15 microg/mL for 1 and 2, respectively), and were not toxic towards the Vero cell line. A regiospecific conversion of the dialdehyde 1 to the lactone 2 proceeded simply under acidic conditions. PMID- 15549668 TI - Potent antibacterial activity of halogenated compounds against antibiotic resistant bacteria. AB - Common Gram-positive clinical pathogens are showing an increasing trend for resistance to conventional antimicrobial agents. New drugs with potent antibacterial activities are urgently needed to remediate this problem. Halogenated compounds isolated from several species of the red algae genus Laurencia were examined for their antibacterial activity against 22 strains of human pathogenic bacteria, 7 strains of which were antibiotic-resistant bacteria. Four phenolic sesquiterpenes and a polybrominated indole showed wide spectra of antibacterial activity against Gram-positive bacteria including methicillin resistant Staphylococcus aureus (MRSA), penicillin-resistant Streptococcus pneumoniae, and vancomycin-resistant Enterococcus faecalis and E. faecium (VRE). In addition, laurinterol and allolaurinterol displayed potent bactericidal activity against three strains of MRSA at 3.13 microg mL(-1), and three strains of vancomycin-susceptible Enterococcus, at 3.13 microg mL(-1) and 6.25 microg mL( 1), respectively. PMID- 15549669 TI - Antifungal activities of essential oils from Thymus quinquecostatus and T. magnus. AB - The antifungal activities of essential oils from Thymus quinquecostatus and T. magnus, which are species native to Korea, were evaluated against seven common pathogenic fungi. Additionally, the effects of the oils together with ketoconazole were tested by the checkerboard titer test. Both of the two Thymus oils showed significant inhibition of the tested fungi, with minimal inhibitor concentrations (MICs) and minimal fungicidal concentrations (MFCs) in the range of 0.04-0.39 mg/mL and 0.19-0.78 mg/mL, respectively. The two Thymus oils, and thymol as well, exhibited synergism with ketoconazole against Trichophyton rubrum, which showed the highest susceptibility to these oils. PMID- 15549670 TI - Trypanocidal activity of a new diterpene from Casearia sylvestris var. lingua. AB - Bioassay-guided fractionation of the hexanic root bark extract of Casearia sylvestris var. lingua led to the isolation of a new clerodane diterpene, whose structure was elucidated as rel-(2 S,5 R,6 R,8 S,9 S,10 R,18 S,19 R)-19-acetoxy 18,19-epoxy-6-hydroxy-18-butanoyloxy-2-(2-methylbutanoyloxy)cleroda-3,13(16), 14 triene by spectroscopic means, including 1D and 2D NMR analyses. This compound showed pronounced activity on Trypanosoma cruzi, the casual agent of Chagas' disease, with minimal inhibitory concentration (MIC) at 0.59 microg/mL. PMID- 15549671 TI - A new cytotoxic phenylbutenoid dimer from the rhizomes of Zingiber cassumunar. AB - A new phenylbutenoid dimer, (+/-)- trans-3-(4-hydroxy-3-methoxyphenyl)-4-[(E)-3,4 dimethoxystyryl]cyclohex-1-ene, was isolated from the rhizomes of Zingiber cassumunar along with the three known compounds, (+/-)- trans-3-(3,4 dimethoxyphenyl)-4-[(E)-3,4-dimethoxystyryl]cyclohex-1-ene, 4-(3,4 dimethoxyphenyl)but-1,3-diene, and 4-(2,4,5-trimethoxyphenyl)but-1,3-diene by bioassay-directed fractionation using the A549 human cancer cell line cytotoxicity assay. Structure of the new compound was elucidated by spectral analysis, including 1D and 2D NMR experiments. PMID- 15549672 TI - Anti-proliferative effects of lichen-derived lipoxygenase inhibitors on twelve human cancer cell lines of different tissue origin in vitro. AB - Lipoxygenases (LOXs) have been implicated in carcinogenesis in various cancer types. In the current study, three structurally different lichen metabolites, protolichesterinic acid (1), lobaric acid (2) and baeomycesic acid (3) were tested for anti-proliferative effects against 12 different human cancer cell lines. All compounds have known in vitro 5-LOX inhibitory activity, and 1 and 2 also inhibit 12-LOX. The activity of the lichen metabolites was compared to that of a specific 5-LOX inhibitor, zileuton (4). The following cancer cell lines were tested: Capan-1, Capan-2 and PANC-1 (all from pancreas), T47-D (breast), PC-3 (prostate), NCI-H1417 (small cell lung), NIH:OVCAR-3 (ovary), AGS (stomach), WiDr (colorectal), HL-60, K-562 and JURKAT (acute promyelocytic, erythro- and T-cell leukemia, respectively). Compound 1 showed the greatest inhibitory effect against all cell lines, with EC50 ranging from 2.4-18.1 microg mL(-1) (7.4-55.8 microM), followed by 2, with EC50 of 15.2 - 65.5 microg mL(-1) (33.2-143.6 microM). The effects of 3 and 4 were of similar orders of magnitude, with EC50 of 28.7 - >80 microg mL(-1) (76.8 - > 213.9 microM) and 12.9 - > 80 microg mL(-1) (50.4 - > 313.7 microM). The dual 5- and 12-LOX inhibitors 1 and to some extent 2 thus exert significant anti-proliferative effects against a variety of human cancer cell lines, while the selective 5-LOX inhibitors 3 and 4 are considerably less active. PMID- 15549673 TI - Effects of green tea on gene expression of hepatic gluconeogenic enzymes in vivo. AB - It has recently been reported that the major green tea polyphenolic constituent, epigallocatechin 3-gallate (EGCG), mimics the cellular effects of insulin including the reductive effect on the gene expression of rate-limiting gluconeogenic enzymes in a cell culture system. We show that administration of green tea that contains EGCG caused a reduction in the level of mRNAs for gluconeogenic enzymes, phosphoenolpyruvate carboxykinase and glucose-6 phosphatase in the mouse liver. EGCG alone was also found to down-regulate the gene expression of these enzymes but not so curcumin or quercetin. The results of this study support the idea that green tea intake may be beneficial in the prevention of diabetes mellitus. PMID- 15549675 TI - Design considerations in the sequential analysis of matched case-control data. AB - A role for sequential test procedures is emerging in genetic and epidemiological studies using banked biological resources. This stems from the methodology's potential for improved use of information relative to comparable fixed sample designs. Studies in which cost, time and ethics feature prominently are particularly suited to a sequential approach. In this paper sequential procedures for matched case-control studies with binary data will be investigated and assessed. Design issues such as sample size evaluation and error rates are identified and addressed. The methodology is illustrated and evaluated using both real and simulated data sets. PMID- 15549676 TI - Survey of the year 2003 commercial optical biosensor literature. AB - In the year 2003 there was a 17% increase in the number of publications citing work performed using optical biosensor technology compared with the previous year. We collated the 962 total papers for 2003, identified the geographical regions where the work was performed, highlighted the instrument types on which it was carried out, and segregated the papers by biological system. In this overview, we spotlight 13 papers that should be on everyone's 'must read' list for 2003 and provide examples of how to identify and interpret high-quality biosensor data. Although we still find that the literature is replete with poorly performed experiments, over-interpreted results and a general lack of understanding of data analysis, we are optimistic that these shortcomings will be addressed as biosensor technology continues to mature. PMID- 15549674 TI - Long-range control of gene expression: emerging mechanisms and disruption in disease. AB - Transcriptional control is a major mechanism for regulating gene expression. The complex machinery required to effect this control is still emerging from functional and evolutionary analysis of genomic architecture. In addition to the promoter, many other regulatory elements are required for spatiotemporally and quantitatively correct gene expression. Enhancer and repressor elements may reside in introns or up- and downstream of the transcription unit. For some genes with highly complex expression patterns--often those that function as key developmental control genes--the cis-regulatory domain can extend long distances outside the transcription unit. Some of the earliest hints of this came from disease-associated chromosomal breaks positioned well outside the relevant gene. With the availability of wide-ranging genome sequence comparisons, strong conservation of many noncoding regions became obvious. Functional studies have shown many of these conserved sites to be transcriptional regulatory elements that sometimes reside inside unrelated neighboring genes. Such sequence-conserved elements generally harbor sites for tissue-specific DNA-binding proteins. Developmentally variable chromatin conformation can control protein access to these sites and can regulate transcription. Disruption of these finely tuned mechanisms can cause disease. Some regulatory element mutations will be associated with phenotypes distinct from any identified for coding-region mutations. PMID- 15549679 TI - Effects of prenatal alcohol exposure on forepaw digit length and digit ratios in rats. AB - Prenatal exposure to alcohol can cause limb and digit defects. Variations in digit ratios in humans are associated with prenatal testosterone exposure. Since prenatal alcohol can reduce testosterone in rats, the effects of prenatal alcohol were measured on rat digit length. Pregnant Sprague-Dawley rats were intubated with 0 g/kg, 4 g/kg, or 6 g/kg of ethanol from Gestational Day 8 (GD8) to GD20. The 0-g/kg group and a nonintubated group served as controls. At postnatal day 31, forepaw digit lengths were measured and digit ratios calculated. Females had smaller digits on both forepaws and higher digit ratios on the right forepaw than males. Rats exposed to 6 g/kg of ethanol had smaller digits than controls on both forepaws and higher digit ratios than controls on the left forepaw. Rat digit ratios differ between the sexes, and prenatal alcohol exposure affects digit ratios. The results are consistent with a perinatal disruption of testosterone levels by alcohol and/or of testosterone's effects on digit length and ratios. An alternate interpretation is consistent with a retinoic acid-mediated effect of alcohol on digit length and ratios. PMID- 15549680 TI - Ontogeny and homology of the paranasal sinuses in Platyrrhini (Mammalia: Primates). AB - The identity and taxonomic distribution of paranasal sinuses among living platyrrhines has remained a contentious issue (e.g., Cave [1967] Am J Phys Anthropol 26:277-288 vs. Hershkovitz [1977] Chicago: University of Chicago Press) largely because the ontogenetic data required for their detection and identification (e.g., Cave [1967]; Maier [2000] Cambridge, UK: Cambridge University Press, 99-132.) were not attainable without sacrificing valuable juvenile and subadult specimens. Non-invasive computed tomography (CT) scanning of ontogenetic series of skulls for 10 platyrrhine genera demonstrates the presence of maxillary and ethmoid sinuses, as well as homologs of the human sphenoid and frontal sinuses. Differences in the latter two sinuses between platyrrhines and hominoids highlight the need for early developmental data in establishing sinus homology. In particular, the identification of homologous recesses in the cartilaginous nasal capsule, from which sinuses later develop, emerges as the critical step. This developmental approach also reveals that the anterior and posterior ethmoid sinuses are each sets of serial homologs, a point which reconciles previous difficulties in establishing sinus homologies across mammalian orders (e.g., Paulli [1900] Gegenbaurs Morphol Jahrb 28:147-178, 179 251, 483-564). PMID- 15549681 TI - Is the integration of heard and seen speech mandatory for infants? AB - For adults and children, speech perception can be significantly influenced by watching a speaker's mouth movements. While recent reports suggest that infants may be able to integrate heard and seen speech, the current research demonstrates that integration is neither as strong or consistent in infants as it is in adults. Three habituation experiments were conducted. In Experiment 1, female (but not male) infants showed evidence of an adult pattern of integration following habituation with an audiovisual /bi/ and testing with audio /bi/-visual /vi/ (perceived as /vi/ by adults). The interpretation of integration was supported, but only in part, by Experiment 2. In Experiment 3, infants were habituated to a mismatched audio /bi/-visual /vi/ combination and tested on concordant /bi/-/bi/ versus /vi/-/vi/ displays. Here, only male infants showed evidence of integration. These results suggest that an initial mechanism supports integration, but that integration is not mandatory for young infants. PMID- 15549682 TI - Role of oxytocin and vasopressin in the transitions of weaning in the rat. AB - Sucklings (18-day-old) and weanlings (35-day-old) were injected icv with oxytocin or its antagonist (both 0.5 microg/1 microl), or vasopressin (1.0 ng/1 microl) or its antagonist (100 ng/1 microl), prior to 4-min observation in a behavioral maze with a sibling in one box and their anesthetized dam in the other. Oxytocin abolished nipple attachment in sucklings, decreased time spent with the dam, and increased self-grooming. The oxytocin antagonist had little influence on behavior. Vasopressin increased self-grooming while its antagonist reduced passive contact with the dam, increased active contact with her, and increased exploration and activity. We conclude that these neuropeptides have diverse roles during weaning, maintaining sucklings' behavior or promoting weaning, and subserving the transition from attachment to the dam to independence from her. We propose that these neurochemicals, and others, mediate the neural, affiliative, and affective changes of weaning, and that the term "weaning" should be understood to encompass these behavioral transitions. PMID- 15549683 TI - Effect of early feeding experience on subsequent prey preference by cuttlefish, Sepia officinalis. AB - Food preferences were investigated in cuttlefish during the first 3 months' posthatching, using choice tests between crabs, shrimps, and young fish. The results showed that without previous feeding experience, cuttlefish preferred shrimps on Day 3. This suggests an innate food preference; however, it was possible to induce a preference for an originally nonpreferred prey item in 3-day old and naive cuttlefish, demonstrating the flexibility of this initial behavioral preference in response to previous individual experience. This preference suggests a learning process involving a form of long-term memory, demonstrated for the first time in juvenile cuttlefish. Until Day 30, juvenile cuttlefish fed exclusively shrimps chose shrimps. This preference probably depends on their previous feeding experience. Finally, it appears that from Day 60, cuttlefish reared on the same restricted diet have a tendency to switch their preference to novel prey items, which diversify their diet. PMID- 15549684 TI - Adrenocortical response to stress in fasted and unfasted artificially reared 12 day-old rat pups. AB - Recent studies have compared artificially reared (AR) rats with dam-reared rats on behavioral outcomes but, despite the fact that they are deprived of their mothers during the stress hyporesponsive period (SHRP), little is known about the effects of AR on the development of the stress response. In this study, the corticosterone (CORT) response to a stressor (saline injection ip) on postnatal Day 12 was assessed in rat pups that had been either dam-reared (DR) or artificially reared since Day 5. In the preceding 24 hr, half the pups in the DR group were maternally deprived (DEP). To control for the food deprivation consequent to maternal deprivation in these groups, half the pups in the AR groups also underwent 24-hr food deprivation (DEP). In the nondeprived condition AR pups did not differ from DR pups on untreated CORT levels or on levels at 1-hr poststress (i.e., all rats demonstrated low levels of CORT characteristic of the SHRP). In contrast, both maternally deprived DR pups and food-deprived AR pups exhibited increased untreated CORT levels as well as a significant increase at 30 min poststress, but CORT elevations were lower in the AR groups than in the DR groups. Thus, long-term maternal deprivation through artificial rearing in rats does not affect the reduced CORT levels and reduced CORT responsiveness associated with the SHRP; however, if animals are food deprived, then all show increased basal CORT levels and a greater CORT response to stress, although this response is lower in AR groups than in DR groups. These results suggest that rat pups artificially reared with adequate nutrition will still exhibit the SHRP. PMID- 15549685 TI - Auditory-visual speech integration by prelinguistic infants: perception of an emergent consonant in the McGurk effect. AB - The McGurk effect, in which auditory [ba] dubbed onto [ga] lip movements is perceived as "da" or "tha," was employed in a real-time task to investigate auditory-visual speech perception in prelingual infants. Experiments 1A and 1B established the validity of real-time dubbing for producing the effect. In Experiment 2, 4 1/2-month-olds were tested in a habituation-test paradigm, in which an auditory-visual stimulus was presented contingent upon visual fixation of a live face. The experimental group was habituated to a McGurk stimulus (auditory [ba] visual [ga]), and the control group to matching auditory-visual [ba]. Each group was then presented with three auditory-only test trials, [ba], [da], and [(delta)a] (as in then). Visual-fixation durations in test trials showed that the experimental group treated the emergent percept in the McGurk effect, [da] or [(delta)a], as familiar (even though they had not heard these sounds previously) and [ba] as novel. For control group infants [da] and [(delta)a] were no more familiar than [ba]. These results are consistent with infants' perception of the McGurk effect, and support the conclusion that prelinguistic infants integrate auditory and visual speech information. PMID- 15549686 TI - Maternal exposure to first-trimester sunshine is associated with increased birth weight in human infants. AB - Two alternative hypotheses have been generated to account for seasonal variation in the birth weight of human infants born in industrialized countries. First, it has been hypothesized that low ambient temperature during the second trimester of gestation decreases birth weight. Second, it has been hypothesized that exposure to bright sunshine during the first trimester increases birth weight. We tested these two hypotheses to determine which, if either, accounted for seasonal variation in birth weight of full-term infants. Birth weight data, collected over a 5-year period, were analyzed as a function of peak and trough sunshine and ambient temperature. Although there was no effect of ambient temperature during any trimester on birth weight, infants whose mothers were exposed to peak sunshine during their first trimester were born significantly heavier than infants whose mothers experienced trough levels of sunshine during the same trimester. Furthermore, infants whose mothers were exposed to trough levels of sunshine during their second and third trimesters were born significantly heavier than infants whose mothers were exposed to peak levels of sunshine during the same trimesters. We hypothesize that high levels of sunshine during early gestation may increase the level of insulin-like growth factor (IGF)-1, facilitating prenatal growth. PMID- 15549687 TI - Leis' conundrum: homology of the clavus of the ocean sunfishes. 2. Ontogeny of the median fins and axial skeleton of Ranzania laevis (Teleostei, Tetraodontiformes, Molidae). AB - One of the most conspicuous characters of the ocean sunfishes, family Molidae, is the punctuation of the body by a deep, abbreviated, caudal fin-like structure extending vertically between the posterior ends of the dorsal and anal fins, termed the clavus by Fraser Brunner. Homology of the clavus has been a matter of debate since the first studies on molid anatomy in the early 1800s. Two hypotheses have been proposed: 1) It is a highly modified caudal fin; 2) It is formed by highly modified elements of the dorsal and anal fins. To resolve this homology issue, we studied the ontogeny of the molid vertebral column and median fins and compared it to that of a less morphologically derived gymnodont (see Part 1 of this study), a member of the family Tetraodontidae. We show that in molids the chorda never flexes during development, that the claval rays form from the posterior ends of the dorsal and anal fins toward the middle, thus closing the gap inward, and that elements of the molid clavus have an identical development and composition as the proximal-middle and distal radials of the regular dorsal and anal fins. We thus conclude that the molid clavus is unequivocally formed by modified elements of the dorsal and anal fin and that the caudal fin is lost in molids. PMID- 15549688 TI - Leis' conundrum: homology of the clavus of the ocean sunfishes. 1. Ontogeny of the median fins and axial skeleton of Monotrete leiurus (Teleostei, Tetraodontiformes, Tetraodontidae). AB - We describe the ontogeny of the axial skeleton and median fins of the Southeast Asian freshwater puffer Monotrete leiurus, based on a reared developmental series. Most elements of the axial skeleton in M. leiurus arise in membrane bone. Only the base of the anterior three neural arches, the base of the hemal arches of the third preural centrum, the neural and hemal arches and spines of the second preural centrum, the parhypural, the two hypural plates, and the single epural are preformed in cartilage. In contrast to most teleosts, the proximal middle radials of the dorsal and anal fins are upright and symmetrical and their distal tips coalesce during development to form a deep band of cartilage, from which the spherical distal radials are spatially separated. PMID- 15549689 TI - Pramipexole in treatment-resistant depression: an extended follow-up. AB - We evaluated the long-term antidepressant safety and response of adjunctive pramipexole, a D2-D3 dopamine agonist, in the course of drug-resistant depression. Twenty-three patients with treatment-resistant major depressive episode (MDE) were followed up after a 16-week pramipexole add-on trial. Pramipexole was added to current treatment with TCA or SSRI, at increasing doses from 0.375-1.500 mg/day. The LIFE scale was administered at baseline of the acute trial, at Weeks 16, 32, and 48. Patients were analyzed for sustained remission (score= <2 at LIFE for at least 8 weeks) and recurrence (after remission score > =3 at LIFE for at least 2 weeks) of depression. Of 23 patients, 12 had major depression and 11 had bipolar depression (16 women; mean age=52.8 years). Mean age of onset and median duration of current MDE were 35.1 years and 6 months, respectively; all subjects had at least two prior MDEs. Mean pramipexole dose was 0.990 mg/day. Median duration of follow-up was 28 weeks. Mean baseline MADRS and CGI-S scores were 33.7+/-8.4 (sd) and 4.6+/-0.8, respectively. Median time to sustained remission from baseline was 10 weeks and overall 60.9% (14/23) of subjects recovered within Week 22. Recurrence of depression occurred in 35.7% (5/14) of remitters after Week 24 and within Week 28 from remission. Although there were no sleep attacks, two cases of hypomania and one case of psychotic mania occurred at Weeks 22, 24, and 30, respectively. Pramipexole augmentation of antidepressant treatment was relatively safe and presumably effective in the long term course of treatment resistant depression. PMID- 15549690 TI - Assessing patient beliefs in a clinical trial of Hypericum perforatum in major depression. AB - Little is known about the beliefs of patients suffering from major depression as to the causes of their illness and effective treatments. This study introduces a new instrument for capturing these beliefs, the Explanatory Model for Depression (EMD) Questionnaire, and explores the beliefs of patients participating in a clinical trial of an alternative medicine, Hypericum perforatum. Although the EMD was originally conceptualized as having five factors pertaining to models of the illness and treatment approaches, the data suggest that patient beliefs are aligned on two factors pertaining to internal and external locus of control. Strong beliefs on either of the EMD locus of control subscales are associated with more severe depression. More importantly, strong beliefs on the external locus of control subscale are associated with less improvement over the 8-week period of observation. These results support the role of patients' beliefs in their recovery from depression and suggest that patients who believe the causes of their depression are outside of their control are less likely to improve over time. It is important to note that beliefs did not mediate the effect of treatment on depression in this study, perhaps because patients were blinded to their treatment condition. Future studies should explore whether patient beliefs have an even greater impact when patients are aware of the treatment they are receiving and can determine whether that treatment is consistent with their beliefs. PMID- 15549691 TI - Neurotoxic regimens of methamphetamine induce persistent expression of phospho-c Jun in somatosensory cortex and substantia nigra. AB - Repeated systemic administration of moderate doses of methamphetamine (mAMPH) can result in neuronal damage. In addition to the prominent damage of forebrain dopamine and serotonin terminals, mAMPH also injures certain non-monoaminergic neuronal somata in the cerebral cortex. In previous studies, we have localized the damaged neurons to the "whisker barrels" in primary somatosensory cortex, reported the time course of their appearance, and found that sensory inputs from the mystacial vibrissae appear to play a crucial role in the mechanism of their injury by mAMPH. One common feature of these studies is that they used a single marker for neuronal injury, the fluorochrome dye Fluoro-Jade, which stains neurons injured by disparate mechanisms. Here we compare mAMPH-induced damage to somatosensory cortical neurons as assessed by Fluoro-Jade and immunohistochemical staining for phospho-c-Jun. A neurotoxic regimen of mAMPH induced phospho-c-Jun positive neurons in both cortical whisker barrels and the substantia nigra. Neurons in the barrel cortex can be sufficiently damaged by mAMPH that they become Fluoro-Jade-positive within 2 hr after the final mAMPH injection. By contrast, phospho-c-Jun immunoreactivity does not appear until 12-24 hr after mAMPH. As reported in an earlier study, unilateral removal of vibrissae prior to mAMPH treatment affords partial protection from injury in the hemisphere contralateral to the vibrissotomy. The vibrissotomized animals show similar decreases in Fluoro-Jade staining and phospho-c-Jun immunoreactivity in the protected hemisphere. Since phospho-c-Jun indicates activation of Jun N-terminal kinase pathways, which have been implicated in apoptosis, we conclude that phospho-c-Jun provides a useful new marker for mAMPH-induced damage to cortical neurons. PMID- 15549692 TI - The FocalPoint System: FocalPoint slide profiler and FocalPoint GS. PMID- 15549693 TI - Probing photonic and optoelectronic structures by apertureless scanning near field optical microscopy. AB - This report presents the Apertureless Scanning Optical Near-Field Microscope as a powerful tool for the characterization of modern optoelectronic and photonic components with sub-wavelength resolution. We present an overview of the results we obtained in our laboratory over the past few years. By significant examples, it is shown that this specific probe microscopy allows for in situ local quantitative study of semiconductor lasers in operation, integrated optical waveguides produced by ion exchange (single channel or Y junction), and photonic structures. PMID- 15549694 TI - Off-axis electron holography of magnetic nanowires and chains, rings, and planar arrays of magnetic nanoparticles. AB - A selection of recent results illustrating the application of off-axis electron holography to the study of magnetic microstructure in closely-spaced nanoparticles and nanowires is reviewed. Examples are taken from the characterization of FeNi nanoparticle chains, Co nanoparticle rings, two dimensional arrays of naturally occurring magnetite crystals in minerals, and single crystalline Co nanowires. Approaches that can be used to separate the magnetic signal of interest from the mean inner potential contribution to the measured holographic phase shift are described, and the spatial and phase resolution that can be achieved are discussed. PMID- 15549695 TI - Application of scanning probe microscopy to the characterization and fabrication of hybrid nanomaterials. AB - Scanning probe microscopy (SPM) is a widely used experimental technique for characterizing and fabricating nanostructures on surfaces. In particular, due to its ability to spatially map variations in materials properties with nanometer spatial resolution, SPM is particularly well suited to probe the subcomponents and interfaces of hybrid nanomaterials, i.e., materials that are made up of distinct nanometer scale components with distinguishable properties. In addition, the interaction of the SPM tip with materials can be intentionally tuned such that local surface modification is achieved. In this manner, hybrid nanostructures can also be fabricated on solid substrates using SPM. This report reviews recent developments in the characterization and fabrication of hybrid nanomaterials with SPM. Specific attention is given to nanomaterials that consist of both organic and inorganic components including individual biomolecules mounted on inorganic substrates. SPM techniques that are particularly well suited for characterizing the mechanical and electrical properties of such hybrid systems in atmospheric pressure environments are highlighted, and specific illustrative examples are provided. This review concludes with a brief discussion of the remaining challenges and promising future prospects for this field. PMID- 15549696 TI - Creating nanoscopic collagen matrices using atomic force microscopy. AB - The atomic force microscope (AFM) is introduced as a biomolecular manipulation machine capable of assembling biological molecules into well-defined molecular structures. Native collagen molecules were mechanically directed into well defined, two-dimensional templates exhibiting patterns with feature sizes ranging from a few nanometers to several hundreds of micrometers. The resulting nanostructured collagen matrices were only approximately 3-nm thick, exhibited an extreme mechanical stability, and maintained their properties over the time range of several months. Our results directly demonstrate the plasticity of biological assemblies and provide insight into the physical mechanisms by which biological structures may be organized by cells in vivo. These nanoscopic templates may serve as platforms on non-biological surfaces to direct molecular and cellular processes. PMID- 15549697 TI - Use of a near-field optical probe to locally launch surface plasmon polaritons on plasmonic waveguides: a study by the finite difference time domain method. AB - We used the finite difference time domain (FDTD) method to study the use of scanning near field optical microscopy (SNOM) to locally excite the nanometric plasmonic waveguides. In our calculation, the light is funneled through a SNOM probe with a sub-wavelength optical aperture and is irradiated on one end of two types of plasmonic waveguides made of 50 nm Au sphere arrays and Au nanowires. The incident light was well localized at one end of the waveguides and consequently propagated toward the other end, due to the excitation of surface plasmon polaritons. We found that the propagation length of the nanosphere array type waveguide varies from 100 to 130 nm depending on the light wavelength, the size of the probe aperture, and the launching heights. Our result shows that reducing the aperture size and using the light of the plasmon resonance wavelength of the nanosphere array could increase the propagation length and, thus, the efficiency of electromagnetic energy transportation through nanosphere arrays. PMID- 15549698 TI - Imaging and analysis of nanowires. AB - We used vapor-liquid-solid (VLS) methods to synthesize discrete single-element semiconductor nanowires and multicomposition nanowire heterostructures, and then characterized their structure and composition using high-resolution electron microscopy (HRTEM) and analytical electron microscopy techniques. Imaging nanowires requires the modification of the established HRTEM imaging procedures for bulk material to take into consideration the effects of finite nanowire width and thickness. We show that high-resolution atomic structure images of nanowires less than 6 nm in thickness have lattice "streaking" due to the finite crystal lattice in two dimensions of the nanowire structure. Diffraction pattern analysis of nanowires must also consider the effects of a finite structure producing a large reciprocal space function, and we demonstrate that the classically forbidden 1/3 [422] reflections are present in the [111] zone axis orientation of silicon nanowires due to the finite thickness and lattice plane edge effects that allow incomplete diffracted beam cancellation. If the operating conditions are not carefully considered, we found that HRTEM image delocalization becomes apparent when employing a field emission transmission electron microscope (TEM) to image nanowires and such effects have been shown to produce images of the silicon lattice structure outside of the nanowire itself. We show that pseudo low dose imaging methods are effective in reducing nanowire structure degradation caused by electron beam irradiation. We also show that scanning TEM (STEM) with energy dispersive X-ray microanalysis (EDS) is critical in the examination of multicomponent nanowire heterostructures. PMID- 15549699 TI - Light emission induced by tunneling electrons from surface nanostructures observed by novel conductive and transparent probes. AB - We have developed an ultrahigh-vacuum low-temperature scanning tunneling microscope (STM) equipped with a near-field optical detection system using novel conductive and optically transparent probes. Tunneling-electron induced photons generated in a nanometer-scale area just under the STM probe can be collected directly into the core of the optical fiber probe within the optical near-field region. Firstly, optical fiber probes coated with indium-tin-oxide thin film are applied to quantitative analysis of p-type GaAs(110) surface, where a decrease of light emission in photon mapping clearly extracts the existence of Zn accepter atoms located at the sub-surface layers. Secondly, in order to enhance the efficiency for inelastic tunneling excitation of a tip-induced plasmon mode, a STM probe coated with an Ag/ITO dual-layer film has been developed and applied to an Ag(111) surface, where photon mapping with a step resolution has been achieved by near-field detection. PMID- 15549700 TI - Developmental changes in the embryo, pronymph, and first molt of the scorpion Centruroides vittatus (scorpiones: buthidae). AB - For the first time the scanning electron microscope was used to compare developmental changes in scorpion embryos and the first and second stadia. In the buthid species of this study, Centruroides vittatus, and all other scorpions, the newborn climb up on their mother's back and remain there without feeding for several days. At this location, they undergo their first molt and in a few days they disperse, fully capable of foraging in the terrestrial environment. The results here support earlier suggestions that the first stadium (pronymph) is a continuation and extension of embryological development. The first molt results in a nymph with exoskeletal features much like those in the adult. In the first molt the metasoma becomes relatively longer, and the sting (aculeus) becomes sharp and functional. The metasomal segments are modified for dorsal flexion and sting use. The embryos and the pronymphs have spiracles that open into an invagination near the posterior margin of flap-like abdominal plates in segments 4-7 of the ventral mesosoma. The second instars have spiracles that lead to book lungs farther anterior in sternites. Tubular legs with cylindrical segments in embryos and pronymphs become more sculptured and oval in the transverse plane. Each leg in the pronymph has a blunt, cup-shaped tip while distal claws (ungues, dactyl) are present in the second instar and subsequent stages. There are some sharp bristles and primordial sensilla in the pronymphs, but the second stadium has adult-like surface features: rows of knobs or granulations (carinae), serrations on the inner surfaces of cheliceral and pedipalpal claws, filtering hairs at the mouthparts, peg sensilla on the pectines, and mechano- and chemoreceptor sensilla on the body and appendages. Scorpion embryos and pronymphs have some structures like fossil scorpions thought to have been aquatic. There is a gradual development of features that appear to be terrestrial adaptations. Evidence is provided for the formation of the sternum from third and fourth leg coxal primordia and possibly from the first abdominal segment. This study is the first to provide evidence for a forward shift of the gonopore along with other structures in the anterior abdomen. PMID- 15549701 TI - Double spermatogenesis in Chelicerata. AB - Sperm dimorphism is a rare phenomenon in Chelicerata. Until now, it was known only from three species of the opilionid genus Siro (Sironidae, Cyphophthalmi). Fertilizing (eusperm) and nonfertilizing spermatozoa (parasperm) develop in the same cyst and are thus sister cells. The fine structure of the spermatozoa of two species has been examined and is compared here. In contrast to Siro rubens, S. duricorius spermatozoa lack an acrosomal complex. Both sperm types produce a transitional process, a more or less modified flagellum, which is later retracted. Hence, the spermatozoa are aflagellate. Eusperm and parasperm of all three species form highly ordered sperm balls that are stored in the deferent duct. Reviewing and adding new results about the sperm dimorphism in this arachnid taxon provides the basis for some considerations of another enigmatic morphological character found in Uropygi and Amblypygi, i.e., the tubular accessory genital glands that show holocrine extrusion. These glands are suggested to represent modified, infertile derivatives of the testis anlage. Their secretion is produced in a way reminiscent of a strongly degenerated spermatogenesis. Consequently, these products may be regarded as strongly degenerated germ cells representing a line of germ cell development, which has been separated very early in spermatogenesis from the usual line leading to fertilizing sperm cells. This further, although less evident, case of probable dichotomous germ cell development is discussed with respect to the controversial phylogenetic-systematic relationships between Uropygi (Thelyphonida and Schizomida), Amblypygi, and Araneae. PMID- 15549702 TI - Coherent nano-area electron diffraction. AB - We describe the new coherent nano-area electron diffraction (NED) and its application for structure determination of individual nanostructures. The study is motivated by the challenge and the general lack of analytical techniques for characterizing nanometer-sized, heterogeneous phases. We show that by focusing electrons on the focal plane of the pre-objective lens using a 3rd condenser lens and a small condense aperture, it is possible to achieve a nanometer-sized highly parallel illumination or probe. The high angular resolution of diffraction pattern from the parallel illumination allows over-sampling and consequently the solution of phase problem based on the recently developed ab initio phase retrieval technique. From this, a high-contrast and high-resolution image can be reconstructed at resolution beyond the performance limit of the image-forming objective lens. The significance of NED for nanostructure characterization will be exemplified by single-wall carbon nanotubes and small metallic clusters. Imaging from diffraction patterns, or diffractive imaging, will be demonstrated using double-wall carbon nanotubes. PMID- 15549703 TI - Nanoscale lead and noble gas inclusions in aluminum: structures and properties. AB - Transmission electron microscopy has been used for structural and physical characterization of nanoscale inclusions of lead and noble gases in aluminum. When the inclusion sizes approach nanoscale dimensions, many of their properties are seen to deviate from similar properties in bulk and in most cases the deviations will increase as the inclusion sizes decrease. Binary alloys of lead and noble gases with aluminum are characterized by extremely low mutual solubilities and inclusions will, therefore, exist as practically pure components embedded in the aluminum matrix. Furthermore, the thermal vacancy mobility in aluminum at and above room temperature is sufficiently high to accommodate volume strains associated with the inclusions thus leading to virtually strain free crystals. The inclusions grow in parallel cube alignment with the aluminum matrix and have a cuboctahedral shape, which reflects directly the anisotropy of the interfacial energies. Inclusions in grain boundaries can have single crystalline or bicrystalline morphology that can be explained from a generalized Wulff analysis such as the xi-vector construction. The inclusions have been found to display a variety of nanoscale features such as high Laplace pressure, size dependent superheating during melting, deviations from the Wulff shape displaying magic size effects, a shape dependence of edge energy, and so on. All these effects have been observed and monitored by TEM using conventional imaging conditions and high-resolution conditions in combination with in-situ analysis at elevated temperatures. PMID- 15549704 TI - Effect of 3,4-di(OH)-cinnamate synthetic derivative on plasma and hepatic cholesterol level and antioxidant enzyme activities in high cholesterol-fed rats. AB - The effect of 3,4-di(OH)-phenylpropionic acid (L-phenylalanine methyl ester) amide (SL-1063), a synthetic derivative of 3,4-di(OH)-cinnamate, on the cholesterol metabolism and antioxidant enzyme system was examined in rats. Diets that included either SL-1063 (0.046%, w/w) or lovastatin (0.02%, w/w) as a supplement, plus 1 g cholesterol/100 g diet were fed to rats ad libitum for 5 weeks. The total plasma cholesterol and triglyceride levels were significantly lowered by the SL-1063 supplement compared to the control group. Meanwhile, the levels of plasma HDL-cholesterol and ratio of HDL-cholesterol/total cholesterol (%) were significantly higher in the SL-1063 group than in the control group. However, the lovastatin supplement did not affect the plasma lipid level. The hepatic cholesterol level and 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity were significantly lowered in the lovastatin group compared to the SL 1063 group; however, the hepatic triglyceride level did not differ among the groups. The activity of hepatic acyl CoA: cholesterol acyltransferase (ACAT), the enzyme that catalyzes hepatic cholesterol esterification, was significantly lower in the lovastatin and SL-1063 groups than in the control group. Furthermore, the SL-1063 supplement elevated the excretion of fecal sterols. As regards the hepatic antioxidant enzyme system, the superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione reductase (GR) activities were all significantly higher in the SL-1063 group compared to the control group, whereas only the GR activity was significantly increased by the lovastatin supplement. No marked difference in the GSH levels and glucose-6-phosphate dehydrogenase (G6PD) activities was observed among the groups. The levels of plasma and hepatic thiobarbituric acid reactive substances (TBARS) were lowered by the SL-1063 supplement compared to the control group. Accordingly, the current results suggest that SL-1063, a synthetic derivative of 3,4-di(OH)-cinnamate, is effective in lowering the plasma lipids and improving the antioxidant enzyme system. PMID- 15549706 TI - Adduct-forming tendencies of cationic triarylmethane dyes with proteins: metabolic and toxicological implications. AB - The formation of colorless adducts by four cationic triarylmethane dyes (TAM(+)s), methyl green (MeG(+)), malachite green (MG(+)), pararosaniline (PR(+)), and crystal violet (CV(+)) was studied spectrophotometrically at 25 degrees C, in 50 mM 3-(N-morpholino)propanesulfonic acid (MOPS) buffer (pH 8), by monitoring the loss in TAM(+) color in the absence and presence of human serum proteins as potential addends. Unfractionated serum caused a rapid bleaching of MeG(+) and MG(+), while PR(+) and CV(+) were unaffected. Sephacryl S200 HR chromatographic screening of the serum revealed two composite peaks of MeG(+) bleaching activity. The major peak (M(r) range, 40,000-130,000) overlapped with and extended on either side of the albumin peak. The minor peak corresponding to ca. 10% of the total MeG(+)-bleaching capacity had M(r) > 230,000. MG(+) bleaching activity dominated the entire chromatographic profile and implicated a multitude of minority proteins with a high capacity to form colorless MG adducts. It is concluded that highly electrophilic TAM(+)s such as MeG(+) and MG(+) must be quantitatively trapped in the form of dye-protein adducts in biological fluids and that the primary in vivo effects (e.g. toxicity) of such dyes most likely arise from ligand-type effects on multiple protein targets. Mechanisms that call for unmodified TAM(+) structure (radical-mediated redox changes, DNA intercalation) may be more relevant to the in vivo impact of dyes such as PR(+) and CV(+) that have a lower tendency to form adducts. PMID- 15549707 TI - Mercury stimulates rat liver glucocorticoid receptor association with Hsp90 and Hsp70. AB - The subject of the present study is the influence of mercury on association of rat liver glucocorticoid receptor (GR) with heat shock proteins Hsp90 and Hsp70. The glucocorticoid receptor heterocomplexes with Hsp90 and Hsp70 were immunopurified from the liver cytosol of rats administered with different doses of mercury. The amounts of co-immunopurified apo-receptor, Hsp90 and Hsp70 were then determined by quantitative Western blotting. The ratio between the amount of heat shock protein Hsp90 or Hsp70 and the amount of apo-receptor within immunopurified heterocomplexes was found to increase in response to mercury administration. On the other hand, the levels of Hsp90 and Hsp70 in hepatic cytosol remained unaltered. The finding that mercury stimulates association of the two heat shock proteins with the glucocorticoid receptor, rendering the cytosolic heat shock protein levels unchanged, suggests that mercury affects the mechanisms controlling the assembly of the receptor heterocomplexes. PMID- 15549708 TI - Possible neuroprotective effects of lecithin and alpha-tocopherol alone or in combination against ischemia/reperfusion insult in rat brain. AB - A close correlation exists between ischemia/reperfusion (I/R)-induced insult and the release of free radicals. Lecithin is a polyunsaturated phosphatidylcholine that corresponds to the phosphatidylcholine molecule. Phosphatidylcholines are high-energy functional and structural elements of all biologic membranes. alpha Tocopherol is the major lipid-soluble chain-breaking antioxidant in the body tissues and effectively protects against neuronal damage. Therefore, we studied the effect of lecithin (300 mg/kg, p.o., 14 days) and alpha-tocopherol (200 mg/kg, p.o., 14 days), alone or in combination, on the brain redox state during I/R. Adult male Wistar rats were subjected to global ischemia by the occlusion of the two carotid arteries 24 h after the last dose of drug administration. Reperfusion was carried out 1 h after induction of ischemia and lasted for another hour. Brain lipid peroxides (MDA) and glutathione (GSH) contents, as well as superoxide dismutase (SOD) and catalase (CAT) activities were assessed. The results showed that I/R elevated brain lipid peroxides content which was accompanied by a reduction in both antioxidant enzyme activities, however, brain GSH level remained unaltered. Lecithin, alpha-tocopherol and their combination restored MDA content, as well as CAT activity with a slight tendency to normalize SOD activity. We conclude that lecithin has a possible neuroprotective effect partly through its antioxidant action which is comparable to that of alpha tocopherol. PMID- 15549710 TI - A mu-class glutathione S-transferase from the marine shrimp Litopenaeus vannamei: molecular cloning and active-site structural modeling. AB - A cDNA clone coding for a mu-class glutathione S-transferase (GST) was isolated from a hepatopancreas cDNA library from the shrimp Litopenaeus vannamei. The deduced amino acid sequence (215 amino acids) has >50% identity to rodents and other mammals mu-class GSTs. Using RT-PCR, the shrimp GST transcript was detected in hepatopancreas, hemocytes, gills, and muscle, but not in pleopods. The shrimp GST sequence was computer modeled and found to fit the classical two-domain GST structure. Domain I, containing the glutathione (GSH) binding site, is more conserved compared to the flexible C-terminal domain II. Residue Q208 appears to be a key to substrate specificity by comparison with mammalian GST mutants. This position is commonly occupied by serine or threonine in mammalian mu-class GSTs, and shrimp Q208 may affect the affinity to substrates like aminochrome or 1,3 dimethyl-2-cyano-1-nitrosoguanidine. This is the first report of molecular cloning and structural modeling of a crustacean GST and provides new insights into the nature of the detoxification response on marine invertebrates. PMID- 15549712 TI - Kinetic properties of turkey pancreatic lipase: a comparative study with emulsified tributyrin and monomolecular dicaprin. AB - Using the classical emulsified system and the monomolecular film technique, we compared several interfacial properties of turkey pancreatic lipase (TPL) and human pancreatic lipase (HPL). TPL, like HPL, presented the interfacial activation phenomenon when vinyl ester was used as substrate. In the absence of colipase and bile salts, using tributyrin emulsion or monomolecular films of dicaprin at low surface pressure, TPL, unlike HPL, hydrolyzes pure tributyrin emulsion as well as dicaprin films maintained at low surface pressures. TPL was also able to hydrolyze triolein emulsion in the absence of any additive and despite the accumulation of long-chain free fatty acids at the interface. The difference of behaviors between TPL and HPL can be explained by the penetration power of each enzyme. The enzyme that presents the maximal pi(c) (TPL) interacts more efficiently with interfaces, and it is not denaturated at high interfacial energy. Turkey pancreatic lipase is more active on rac-dicaprin than HPL; a maximal ratio of 9 was found between the catalytic activities of the two lipases measured at their surface pressure optima (20 mN m(-1)). A kinetic study on the surface pressure dependency, stereospecificity, and regioselectivity of TPL was performed using enantiopure diglyceride (1,2-sn-dicaprin and 2,3-sn-dicaprin) and a prochiral isomer (1,3-dicaprin) that were spread as monomolecular films at the air-water interface. At low surface pressure (15 mN m(-1)), TPL acts preferentially on primary carboxylic ester groups of the diglyceride isomers (1,3 dicaprin), but at high surface pressure (23 mN m(-1)), this enzyme prefers both adjacent ester groups of the diglyceride isomers (1,2-sn-dicaprin and 2,3-sn dicaprin). HPL prefers adjacent ester groups of the diglyceride isomers (1,2-sn dicaprin and 2,3-sn-dicaprin). Furthermore, TPL was found to be markedly stereospecific for the sn-1 position of the 1,2-sn-enantiomer of dicaprin at low surface pressure (15 mN m(-1)), while at high surface pressure (23 mN m(-1)), this lipase presents a stereopreference for the sn-3 position of the 2,3-sn enantiomer of dicaprin. HPL is stereospecific for the sn-1 position of the 1,2-sn enantiomer of dicaprin both at 15 and 23 mN m(-1). PMID- 15549711 TI - Scoparia dulcis, a traditional antidiabetic plant, protects against streptozotocin induced oxidative stress and apoptosis in vitro and in vivo. AB - Oxidative stress is implicated in the pathogenesis of diabetic complications. The experiments were performed on normal and experimental male Wistar rats treated with Scoparia dulcis plant extract (SPEt). The effect of SPEt was tested on streptozotocin (STZ) treated Rat insulinoma cell lines (RINm5F cells) and isolated islets in vitro. Administration of an aqueous extract of Scoparia dulcis by intragastric intubation (po) at a dose of 200 mg/kg body weight significantly decreased the blood glucose and lipid peroxidative marker thiobarbituric acid reactive substances (TBARS) with significant increase in the activities of plasma insulin, pancreatic superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in streptozotocin diabetic rats at the end of 15 days treatment. Streptozotocin at a dose of 10 mug/mL evoked 6-fold stimulation of insulin secretion from isolated islets indicating its insulin secretagogue activity. The extract markedly reduced the STZ-induced lipidperoxidation in RINm5F cells. Further, SPEt protected STZ-mediated cytotoxicity and nitric oxide (NO) production in RINm5F cells. Treatment of RINm5F cells with 5 mM STZ and 10 mug of SPEt completely abrogated apoptosis induced by STZ, suggesting the involvement of oxidative stress. Flow cytometric assessment on the level of intracellular peroxides using fluorescent probe 2'7'-dichlorofluorescein diacetate (DCF-DA) confirmed that STZ (46%) induced an intracellular oxidative stress in RINm5F cells, which was suppressed by SPEt (21%). In addition, SPEt also reduced (33%) the STZ-induced apoptosis (72%) in RINm5F cells indicating the mode of protection of SPEt on RIN m5Fcells, islets, and pancreatic beta-cell mass (histopathological observations). Present study thus confirms antihyperglycemic effect of SPEt and also demonstrated the consistently strong antioxidant properties of Scoparia dulcis used in the traditional medicine. PMID- 15549713 TI - Synthesis and absolute configuration assignment of 5-amino-1,3,5-triphenyl pentane-1,3-diol stereoisomers. AB - Starting from 2,4,6-triphenylpyrylium perchlorate, 5-amino-1,3,5-triphenyl pentane-1,3-diol stereoisomers 4 were obtained in a simple two-step synthesis: reaction with hydroxylamine, and reduction with LAH of the resulting 2 isoxazoline ketone derivative 2. The eight stereoisomers of 4 were separated in a single shot on a chiral stationary phase cellulose tris(3,5 dimethylphenylcarbamate) (Chiralcel OD-H). The absolute configuration of the title compounds, intermediate 2-isoxazoline ketone 2 and isoxazoline alcohol derivative 3 were determined using a combination of diastereoselective synthesis, affiliation of the sign in chemical interconversion method, and X-ray determination. 2-Isoxazoline ketone 2 enantiomers and isoxazoline alcohol 3 enantiomers were obtained by chiral HPLC on Chiralpak AD column. 2-Isoxazoline ketone 2 enantiomers can be racemized via a retro Michael addition. PMID- 15549715 TI - That certain age. PMID- 15549714 TI - Enlarged and prominent nucleoli may be indicative of MYCN amplification: a study of neuroblastoma (Schwannian stroma-poor), undifferentiated/poorly differentiated subtype with high mitosis-karyorrhexis index. AB - BACKGROUND: According to the International Neuroblastoma Pathology Classification, neuroblastomas exhibiting MYCN amplification (A-MYCN) have unique histologic features-namely, undifferentiated/poorly differentiated subtype with a high mitosis-karyorrhexis index (U/PD-H). Nonetheless, certain tumors possessing these histologic characteristics contain a nonamplified MYCN gene (NA-MYCN). METHODS: The clinical characteristics of patients from the Children's Cancer Group (CCG) 3881 and 3891 studies who had neuroblastoma, U/PD-H, exhibiting A MYCN (n=68) or NA-MYCN (n=33) were investigated. The histologic and cytologic features of tumors (A-MYCN, n=62; NA-MYCN, n=28) filed at the Pathology Reference Laboratory, Department of Pathology and Laboratory Medicine, Childrens Hospital Los Angeles, were reviewed, and nucleolar areas in undifferentiated neuroblastic cells were evaluated using image analysis methods. RESULTS: All 68 patients whose tumors exhibited A-MYCN had disease that was in an advanced clinical stage (Stage III or IV); 89.7% of these patients were diagnosed between ages 0.5 and 3.5 years, and 67 of the 68 had been treated with the high-risk protocol in the CCG 3891 study. Children whose tumors exhibited NA-MYCN were evenly distributed across all age groups; 30 of these 33 children had advanced-stage disease, and 26 had been treated with a high-risk protocol. The prognosis associated with A-MYCN (event free survival [EFS], 15.7%; overall survival [OS], 22.2%) was significantly poorer than the prognosis associated with NA-MYCN (EFS, 56.1%; OS, 69.3%). The lone histologic/cytologic difference between tumors exhibiting A-MYCN and tumors exhibiting NA-MYCN involved nucleolar appearance. Neuroblastic cells in tumors exhibiting A-MYCN were characterized by the presence of 1 or more large, prominent nucleoli, and the mean nucleolar area was significantly greater in the 18 tumors exhibiting A-MYCN that were assessed (7.63 microm2) than in the 16 tumors exhibiting NA-MYCN that were assessed (5.53 microm2; P=0.004). CONCLUSIONS: Neuroblastomas, U/PD-H, were found to vary in terms of molecular background and clinical behavior. The results of the current study indicate that nucleolar enlargement in neuroblastic cells may be a sign of MYCN amplification. PMID- 15549716 TI - From discretion to disagreement: explaining disparities in judges' pretrial decisions. AB - Judges are afforded considerable discretion in decision-making. Through their exercise of discretion, judges construct society's notion of crime and justice. This study examined 61 lay judges' bail decision-making in the English criminal justice system. The law states that in particular cases decisions to grant bail or remand in custody should be based on the risk of a defendant absconding, offending, or obstructing justice while on bail. However, there is little guidance on how these judgments should be made and how they should affect decisions. It was found that judges varied (disagreed) in their risk judgments and decisions on the same set of simulated cases. The extent of judicial disagreement differed across cases, and the source of disagreement in decisions lay in the variability of judges' earlier risk judgments. The paper discusses how judicial disagreement may be reduced. PMID- 15549717 TI - Olfactory morphology of carcharhinid and sphyrnid sharks: does the cephalofoil confer a sensory advantage? AB - Many hypotheses have been advanced to explain the adaptive significance of the sphyrnid cephalofoil, including potential advantages of spacing the olfactory organs at the distal tips of the broad surface. We employed comparative morphology to test whether the sphyrnid cephalofoil provides better stereo olfaction, increases olfactory acuity, and samples a greater volume of the medium compared to the situation in carcharhiniform sharks. The broadly spaced nares provide sphyrnid species with a significantly greater separation between the olfactory rosettes, which could lead to an enhanced ability to resolve odor gradients. In addition, most sphyrnid species possess prenarial grooves that greatly increase the volume of water sampled by the nares and thus increase the probability of odorant encounter. However, despite a much greater head width, and a significantly greater number of olfactory lamellae, scalloped hammerhead sharks do not possess a greater amount of olfactory epithelial surface area than the carcharhiniform sandbar sharks. Therefore, sphyrnid sharks might not possess any greater olfactory acuity than carcharhinids. Despite this, there are clear olfactory advantages to the cephalofoil head morphology that could have led to its evolution, persistence, and diversification. persistence, and diversification. PMID- 15549718 TI - Cranial anatomy of the extinct amphisbaenian Rhineura hatcherii (Squamata, Amphisbaenia) based on high-resolution X-ray computed tomography. AB - The fossilized skull of a small extinct amphisbaenian referable to Rhineura hatcherii Baur is described from high-resolution X-ray computed tomographic (HRXCT) imagery of a well-preserved mature specimen from the Brule Formation of Badlands National Park, South Dakota. Marked density contrast between bones and surrounding matrix and at bone-to-bone sutures enabled the digital disarticulation of individual skull elements. These novel visualizations provide insight into the otherwise inaccessible three-dimensionally complex structure of the bones of the skull and their relationships to one another, and to the internal cavities and passageways that they enclose. This study corrects several previous misidentifications of elements in the rhineurid skull and sheds light on skull construction generally in "shovel-headed" amphisbaenians. The orbitosphenoids in R. hatcherii are paired and entirely enclosed within the braincase by the frontals; this is in contrast to the condition in many extant amphisbaenians, in which a large azygous orbitosphenoid occupies a topologically distinct area of the skull, closing the anterolateral braincase wall. Rhineura hatcherii retains a vestigial jugal and a partially fused squamosal, both of which are absent in many extant species. Sculpturing on the snout of R. hatcherii represents perforating canals conveying sensory innervation; thus, the face of R. hatcherii receives cutaneous innervation to an unprecedented degree. The HRXCT data (available at www.digimorph.org) corroborate and extend previous hypotheses that the mechanical organization of the head in Rhineura is organized to a large degree around its burrowing lifestyle. PMID- 15549719 TI - Comparative histological and immunohistochemical study of sea star tube feet (Echinodermata, Asteroidea). AB - Adhesion in sea stars is the function of specialized structures, the tube feet or podia, which are the external appendages of the water-vascular system. Adhesive secretions allow asteroid tube feet to perform multiple functions. Indeed, according to the sea star species considered, the tube feet may be involved in locomotion, fixation, or burrowing. Different tube foot shapes usually correspond to this variety of function. In this study, we investigated the variability of the morphology of sea star tube feet as well as the variability of the composition of their adhesive secretions. This second aspect was addressed by a comparative immunohistochemical study using antibodies raised against the adhesive material of the forcipulatid Asterias rubens. The tube feet from 14 sea star species representing five orders and 10 families of the Class Asteroidea were examined. The histological study revealed three main tube foot morphotypes, i.e., knob-ending, simple disc-ending, and reinforced disc-ending. Analysis of the results suggests that tube foot morphology is influenced by species habitat, but within limits imposed by the evolutionary lineage. In immunohistochemistry, on the other hand, the results were very homogeneous. In every species investigated there was a very strong immunolabeling of the adhesive cells, independently of the taxon considered, of the tube foot morphotype or function, or of the species habitat. This indicates that the adhesives in all the species considered are closely related, probably sharing many identical molecules or, at least, many identical epitopes on their constituents. PMID- 15549720 TI - Larval development of Lynceus brachyurus (Crustacea, Branchiopoda, Laevicaudata): redescription of unusual crustacean nauplii, with special attention to the molt between last nauplius and first juvenile. AB - The larval development of "conchostracans" has received only scattered attention. Here I present the results of a study on the larval (naupliar) development and the metamorphosis of Lynceus brachyurus, a member of the bivalved branchiopod order the Laevicaudata. Lynceus brachyurus is the only species of the "Conchostraca" in Denmark. The phylogenetic position of the Laevicaudata has traditionally been a source of controversy, and this study does not solve the question completely. This work focuses on features potentially important for phylogeny. The general appearance of the larvae of L. brachyurus has been known for more than a century and a half, and some of its unique features include a large, larval dorsal shield; a huge, plate-like labrum; and a pair of immovable, horn-like antennules. However, many details relating to limb morphology, potentially important for phylogeny, have not been studied previously. Based on size categories, five or six larval stages can be recognized. The larvae approximately double their length and width during development (length: 230-520 microm). Most morphological features stay largely unchanged during development, but the antennal coxal masticatory spines are significant exceptions: they become bifid after one of the first molts. In all larval stages only the antennae and the mandibles actively move. In late naupliar stages the trunk limbs become visible as rows of laterally placed, undeveloped, and still immovable lobes. Swimming is performed by the antennae, whereas the mandibles appear to be involved mainly in feeding, as in other branchiopod larvae. The last naupliar stage undergoes a small metamorphosis to the first juvenile stage, the details of which in part were studied by following the premolt juvenile condition through the cuticle of the last stage nauplius. Among other changes there is a characteristic change in the shape and morphology of the univalved dorsal naupliar shield to a bivalved juvenile carapace. The general morphologies of the antennae and the mandibles are very similar to those of other branchiopod larvae and fall well within the "branchiopod naupliar feeding apparatus" recognized as a branchiopod synapomorphy by Olesen (2003), but some specific features shared with the larvae of other "conchostracans" are also identified. These special "conchostracan" features include: 1) a similar antennular setation; 2) a similar comb-like setulation of the bifid antennal coxal processes; and 3) mandibular palpsetae with setules condensed. In light of recent suggestions concerning branchiopod phylogeny (Cyclestheria as a sister group to the Cladocera), these similarities probably do not support a monophyletic "Conchostraca" but rather are symplesiomorphies of this taxon. A final decision must await a phylogenetic analysis of a more complete set of characters. PMID- 15549721 TI - Diversity of pectoral fin structure and function in fishes with labriform propulsion. AB - Aquatic propulsion generated by the pectoral fins occurs in many groups of perciform fishes, including numerous coral reef families. This study presents a detailed survey of pectoral fin musculoskeletal structure in fishes that use labriform propulsion as the primary mode of swimming over a wide range of speeds. Pectoral fin morphological diversity was surveyed in 12 species that are primarily pectoral swimmers, including members of all labroid families (Labridae, Scaridae, Cichlidae, Pomacentridae, and Embiotocidae) and five additional coral reef fish families. The anatomy of the pectoral fin musculature is described, including muscle origins, insertions, tendons, and muscle masses. Skeletal structures are also described, including fin shape, fin ray morphology, and the structure of the radials and pectoral girdle. Three novel muscle subdivisions, including subdivisions of the abductor superficialis, abductor profundus, and adductor medialis were discovered and are described here. Specific functional roles in fin control are proposed for each of the novel muscle subdivisions. Pectoral muscle masses show broad variation among species, particularly in the adductor profundus, abductor profundus, arrector dorsalis, and abductor superficialis. A previously undescribed system of intraradial ligaments was also discovered in all taxa studied. The morphology of these ligaments and functional ramifications of variation in this connective tissue system are described. Musculoskeletal patterns are interpreted in light of recent analyses of fin behavior and motor control during labriform swimming. Labriform propulsion has apparently evolved independently multiple times in coral reef fishes, providing an excellent system in which to study the evolution of pectoral fin propulsion. PMID- 15549722 TI - Direct quantitation of poly(ADP-ribose) polymerase (PARP) activity as a means to distinguish necrotic and apoptotic death in cell and tissue samples. PMID- 15549723 TI - Breast cancer survivors involved in vigorous team physical activity: psychosocial correlates of maintenance participation. AB - Physical activity is increasingly being promoted as a means to achieve both physical and psychological benefits for cancer survivors. For women with breast cancer, one sport growing in popularity is dragon boating. The purpose of the present investigation was to examine the psychosocial correlates of dragon boat participation over the course of a season. Six crews completed the baseline (early-season) assessment (n = 109) and late-season assessments (n = 56). The self-report questionnaire completed at both time points included an assessment of the theory of planned behaviour variables, quality of life, cohesion, and physical activity levels. A prospective examination of the TPB variables revealed attitude at early season as the only significant predictor of behavioural intentions 12 weeks later at late season (R2 adjusted = 0.27, p < 0.001). Overall, the group environment was cohesive at a level similar to that for female sport teams among the asymptomatic population. As well, participants' health related quality of life was similar to normal, healthy women of similar age for both mental and physical health. PMID- 15549724 TI - Site-specific in vivo labeling of proteins for NMR studies. PMID- 15549725 TI - Direct observation of anion-mediated translocation of fluorescent oligoarginine carriers into and across bulk liquid and anionic bilayer membranes. AB - The recent hypothesis that counteranion-mediated dynamic inversion of charge and solubility might contribute to diverse functions of oligoarginines in biomembranes was tested with two fluorescently labelled oligomers, FL-R(8), one of the most active cell-penetrating peptides, and its longer version, FL-R(16). We report evidence for counteranion-mediated phase transfer from water into bulk chloroform and anionic lipid-bilayer membranes as well as reverse-phase transfer from bulk chloroform and across intact lipid-bilayer membranes into water. The differences found between FL-R(8) and FL-R(16) with regard to location in the bilayer and reverse-phase transfer from bulk and lipid-bilayer membranes into water implied that the reported results may be relevant for biological function. PMID- 15549726 TI - Hoechst 33258 as a pH-sensitive probe to study the interaction of amine oxide surfactants with DNA. AB - The use of Hoechst 33258 (HO) as a fluorescent probe to characterize the interactions between DNA and pH-sensitive amphiphiles is discussed. In the case of amine oxide amphiphiles dodecyldimethylamine oxide (DDAO) and p dodecyloxybenzyldimethylamine oxide (pDoAO), the decrease in fluorescence emission, which signals DNA-amphiphile association, is accompanied by a large hypsochromic shift in the emission maximum of the bound probe; this eventually reaches a value characteristic of HO in a neutral or slightly basic environment. These findings are compared to results obtained by using the more common ethidium bromide (EB) probe, which shows no such shift. Circular dichroism and fluorescence depolarization experiments indicate that fluorescence emission only occurs from the DNA-bound probe, and the observed shift in emission maximum when using HO as a fluorescent probe is due to a variation in the local pH in the vicinity of the probe. PMID- 15549727 TI - Interleukin 1 plays a major role in the development of Th2-mediated immunity. AB - Expulsion of the gastrointestinal nematode Trichuris muris is mediated by a T helper (Th)2-type response, involving interleukin (IL)-4, IL-9 and IL-13. Here, we show that Th2 response-associated resistance is dependent on the presence of IL-1alpha and IL-1beta. When lymph node cells from naive IL-1alpha- or IL-1beta deficient mice were subjected to Th2 polarization in vitro, they failed to polarize in the presence of IL-4 alone, but required the addition of exogenous IL 1alpha or IL-1beta. Furthermore, we demonstrate that both IL-1alpha- and IL-1beta deficient mice are susceptible to chronic T. muris infection and that the inability to expel the worms is associated with a defect in the development of a Th2 response in the mesenteric lymph nodes. These results provide the first demonstration of the critical role of IL-1 in regulating Th2 responses during gastrointestinal nematode infection. PMID- 15549728 TI - Development of an anti-IL-12 p40 auto-vaccine: protection in experimental autoimmune encephalomyelitis at the expense of increased sensitivity to infection. AB - IL-12 and IL-23, which share the IL-12 p40 subunit, have been ascribed central roles in many autoimmune disorders. We describe here an anti-IL-12 (alphaIL-12) auto-vaccine that potentially blocks both factors in vivo. Immunization of mice with mouse IL-12 coupled to OVA or Pan DR epitope (PADRE) peptide induced Ab directed against the IL-12 p40 subunit, which prevented IFN-gamma production in response to IL-12 administration in vivo. Experimental autoimmune encephalomyelitis, an IL-23-dependent disease model, induced in SJL mice with a proteolipid protein (PLP) peptide was almost undetectable after alphaIL-12 vaccination. Myelin oligodendrocyte glycoprotein (MOG)-induced disease in C57BL/6 mice was also significantly inhibited. This protection correlated with inhibited Th1 cytokine responses in vitro and with an increase in the IgG1/IgG2a anti-PLP Ab balance. Detrimental consequences of alphaIL-12 vaccination were evaluated in C57BL/6 mice infected with Leishmania major (L.m.). While delayed-type hypersensitivity (DTH) suppression and immunoglobulin as well as interleukin production patterns reflected a major shift toward a Th2-type response, L.m. growth was still significantly retarded as compared to that seen in susceptible BALB/c mice. However, vaccinated animals ultimately failed to control parasite expansion. These results suggest that some chronic autoimmune diseases may benefit from alphaIL-12 vaccination at the expense of reduced, but not completely abrogated, cell-mediated immunity. PMID- 15549729 TI - The molecular requirements for LAT-mediated differentiation and the role of LAT in limiting pre-B cell expansion. AB - Successful recombination of the heavy-chain locus in developing B cells results in the expression of the pre-BCR, which induces the proliferation and expansion of pre-B cells. To avoid uncontrolled proliferation, pre-BCR signals transmitted via the adaptor protein SLP-65 (SH2-domain-containing leukocyte protein of 65 kDa) lead to the down-regulation of pre-BCR expression and to pre-B cell differentiation. Here, we show that, similarly to SLP-65, the adaptor protein LAT (linker for activation of T cells) limits pre-B cell proliferation and reduces the potential of a tumorgenic pre-B cell line to develop leukemia in immune deficient mice. We further show that the four distal tyrosines are required for LAT activity in pre-B cells. Mutation at Y136 completely abolishes LAT activity, whereas single point-mutations at Y175, Y195 or Y235 impair, but do not block, LAT-induced pre-B cell differentiation. As LAT is also expressed in human pre-B cells, our results suggest that LAT cooperates with SLP-65 to promote the differentiation and control the proliferation of both murine and human pre-B cells. PMID- 15549730 TI - A synthetic HIV-1 Tat protein breaches the skin barrier and elicits Tat neutralizing antibodies and cellular immunity. AB - The HIV-1 Tat protein plays a critical role in the pathogenesis of HIV and has been considered as a candidate vaccine antigen. In an effort to design a non invasive vaccination strategy against HIV-1 that stimulates the induction of systemic and mucosal immune responses, we studied the transcutaneous delivery of a synthetic Tat protein using cholera toxin as an adjuvant. Following immunization of BALB/c mice with various doses of Tat, IgG and IgA antibody responses were measured in the serum and vaginal washes, respectively. Serum antibodies predominantly recognized the N-terminal and basic functional domains of the protein and exhibited neutralizing capacity against Tat-driven transactivation. Transcutaneous immunization also elicited potent cellular immune responses against Tat and the secretion of high levels of IL-2, IFN-gamma and IL 6. These findings demonstrate for the first time that by using a simple and safe immunization procedure, a synthetic Tat protein can elicit potentially protective immune responses. Transcutaneous immunization may be advantageous for the non invasive delivery of other HIV candidate vaccine antigens. PMID- 15549731 TI - IREM-1 is a novel inhibitory receptor expressed by myeloid cells. AB - Using a three-hybrid strategy, we have identified a novel cell surface molecule which interacts with the Src homology 2 (SH2) domains of SH2 domain-containing protein tyrosine phosphatase 1 (SHP-1), termed "immune receptor expressed on myeloid cells 1" (IREM-1). The full-length cDNA coding for a polypeptide of 290 amino acids presents an extracellular single V-type Ig domain, a transmembrane region and a cytoplasmic tail with five tyrosine residues, two of which are in the context of an immunoreceptor tyrosine-based inhibitory motif. Moreover, cDNA encoding for three other splicing forms of IREM-1, named IREM-1 splice variant (Sv)1, Sv2 and Sv3 were cloned by reverse transcription (RT)-PCR. The gene encoding for IREM-1 contains nine exons, is located on human chromosome 17 (17q25.1) and is homologous to previously identified molecules termed CMRF-35 and IRp60. RT-PCR, northern blot and FACS analysis with specific monoclonal antibodies indicated that IREM-1 is expressed on monocytes, granulocytes, and myeloid leukemia cell lines. Western blot analysis confirmed the recruitment of SHP-1 to IREM-1 and demonstrated that phosphotyrosine residue 205 is the main docking site for this interaction. Finally, cross-linking of IREM-1 results in the inhibition of FcRepsilon-induced activation. Our results indicate that IREM-1 is a novel inhibitory receptor of the Ig superfamily in myeloid cells. PMID- 15549732 TI - Inhibition of mast cells by interleukin-10 gene transfer contributes to protection against acute myocarditis in rats. AB - Progression of acute myocarditis involves a variety of inflammatory events. Mast cells have been implicated as the source of various cytokines, chemokines and histamine in acute inflammation and fibrosis. Interleukin (IL)-10 has well-known immunomodulatory actions that are exerted during the recovery phase of myocarditis. In this study, 9-week-old male Lewis rats were immunized with cardiac myosin. A plasmid vector expressing mouse IL-10 cDNA (800 mug per rat) was then transferred three times (7, 12 and 17 days after immunization) into the tibialis anterior muscles of the rats by electroporation. Microscopic examination of mast cells was carried out on toluidine blue-stained transverse sections of the mid ventricles. Mouse IL-10 gene transfer significantly reduced mast cell density, cardiac histamine concentration and mast cell growth, and prevented mast cell degranulation. Furthermore, improvement in both myocardial function and the overall condition of the rats was evident from the reduction in the heart weight to-body weight ratio and inflammatory infiltration as well as improvement in hemodynamic and echocardiographic parameters. These findings suggest that IL-10 gene transfer by electroporation protected against myocarditis via mast cell inhibition. PMID- 15549733 TI - Distal regulatory elements play an important role in regulation of the human IL-5 gene. AB - Eosinophil infiltration of the lung is a feature of both allergic and nonallergic asthma, and IL-5 is the key cytokine regulating the production and activation of these cells. Despite many studies focusing on the IL-5 promoter in both humans and mice there is as yet no clear picture of how the IL-5 gene is regulated. The aim of this study was to determine if distal regulatory elements contribute to appropriate regulation of the human IL-5 (hIL-5) gene. Activity of the -507/+44 hIL-5 promoter was compared to expression of the endogenous IL-5 gene in PER-117 T cells. The IL-5 promoter was not sufficient to reproduce a physiological pattern of IL-5 expression. Further, functional analysis of the 5' and 3' intergenic regions revealed a number of novel regulatory elements. We have identified a conserved enhancer located approximately 6.2 kb upstream of the hIL 5 gene. This region contains two potential GATA-3-binding sites and increases expression from the hIL-5 promoter by up to ninefold. PMID- 15549734 TI - Expression of inhibitory KIR is confined to CD8+ effector T cells and limits their proliferative capacity. AB - A subset of effector/memory CD8(+) T cells expresses natural killer cell receptors (NKR). Expression of inhibitory NKR at that stage of T cell differentiation is poorly understood. Interestingly, recent studies in mice indicated that transgenic expression of an inhibitory NKR induced the accumulation of memory T cells by inhibiting activation-induced cell death (AICD). To further understand the role of inhibitory NKR on T cells, we characterized the subset of human peripheral T cells expressing the inhibitory NKR, CD158b, and studied the modulation of antigen-driven T cell expansion by an endogenous inhibitory NKR. We found that CD158b expression was confined to a population of CD8(+)TCRalphabeta(+) effector T cells as defined by a CD45RA(+)CCR7(-) phenotype and high constitutive expression of granzyme B1. Few cells expressed the activating form CD158j in the absence of CD158b. Functionally, engagement of CD158b by MHC ligands diminished early TCR signaling, as well as AICD. However, the reduced AICD did not rescue cells for proliferation, since T cell expansion in the presence of CD158b triggering was impaired. Expression of inhibitory NKR on effector CD8(+) T cells may explain in part the poor replicative capacity of T cells at that stage of differentiation. PMID- 15549735 TI - Vaccination with autoantigen protects against aggregated beta-amyloid and glutamate toxicity by controlling microglia: effect of CD4+CD25+ T cells. AB - Neurodegenerative diseases differ in etiology but are propagated similarly. We show that neuronal loss caused by intraocular injection of aggregated beta amyloid was significantly greater in immunodeficient mice than in normal mice. The neurodegeneration was attenuated or augmented by elimination or addition, respectively, of naturally occurring CD4(+)CD25(+) regulatory T cells (Treg). Vaccination with retina-derived antigens or with the synthetic copolymer glatiramer acetate (Copolymer-1, Cop-1), but not with beta-amyloid, reduced the ocular neuronal loss. In mouse hippocampal slices, microglia encountering activated T cells overcame the cytotoxicity of aggregated beta-amyloid. These findings support the concept of "protective autoimmunity", show that a given T cell-based vaccination is protective at a particular site irrespective of toxicity type, and suggest that locally activated T cells induce a microglial phenotype that helps neurons withstand the insult. Alzheimer's and other neurodegenerative diseases might be arrested or retarded by vaccination with Cop 1 or related compounds or by treatment with compounds that weaken Treg suppression. PMID- 15549736 TI - The dithieno[3,2-b:2',3'-d]phosphole system: a novel building block for highly luminescent pi-conjugated materials. PMID- 15549737 TI - Dramatic enhancement of catalytic activity in an ionic liquid: highly practical Friedel-Crafts alkenylation of arenes with alkynes catalyzed by metal triflates. PMID- 15549738 TI - Molecular lead clusters--from unexpected discovery to rational synthesis. PMID- 15549739 TI - Organocatalyzed conjugate umpolung of alpha,beta-unsaturated aldehydes for the synthesis of gamma-butyrolactones. PMID- 15549740 TI - Butenyl-substituted alkaline-earth metallocenes: a first step towards olefin complexes of the alkaline-earth metals. PMID- 15549742 TI - Polycationic Mn12 single-molecule magnets as electron reservoirs with S > 10 ground states. PMID- 15549743 TI - Rapidly initiating ruthenium olefin-metathesis catalysts. PMID- 15549744 TI - Nanozymes: gold-nanoparticle-based transphosphorylation catalysts. PMID- 15549745 TI - A general route to macroscopic hierarchical 3D nanowire networks. PMID- 15549746 TI - A catalytic Langmuir film as a model for heterogeneous and homogeneous catalytic processes. PMID- 15549747 TI - Catalytic asymmetric mercuriocyclization of gamma-hydroxy-cis-alkenes. PMID- 15549748 TI - Nickel-catalyzed cross-coupling reaction of alkyl halides with organozinc and Grignard reagents with 1,3,8,10-tetraenes as additives. PMID- 15549750 TI - [LAl(mu-S3)2AlL]: a homobimetallic derivative of the sulfur crown S8. PMID- 15549749 TI - Nitroxide-mediated controlled free-radical emulsion polymerization of styrene and n-butyl acrylate with a water-soluble alkoxyamine as initiator. PMID- 15549751 TI - Preparation of [LAl(mu-S)2MCp2] (M = Ti, Zr) from the structurally characterized lithium complexes [[LAl(SH)[SLi(thf)2]]2] and [[LAl[(SLi)2(thf)3]]2]2 THF. PMID- 15549753 TI - Biomimetic engineering of carbon nanotubes by using cell surface mucin mimics. PMID- 15549754 TI - Probing the influence of O2 on photoinduced reversible electron transfer in perylenediimide-triphenylamine-based dendrimers by single-molecule spectroscopy. PMID- 15549755 TI - An oriented 1D coordination/organometallic dimetallic molecular wire with Ag-Pd metal-metal bonds. PMID- 15549756 TI - Asymmetric synthesis of diarylmethyl amines by rhodium-catalyzed asymmetric addition of aryl titanium reagents to imines. PMID- 15549757 TI - A gallium-coated gold cluster. PMID- 15549758 TI - Templating open- and closed-chain structures around metal complexes of macrocycles. PMID- 15549759 TI - Reversible photo-switching of the magnetization of iron oxide nanoparticles at room temperature. PMID- 15549760 TI - Direct oxidative Heck cyclizations: intramolecular Fujiwara-Moritani arylations for the synthesis of functionalized benzofurans and dihydrobenzofurans. PMID- 15549761 TI - Changes in the conductance of single peptide molecules upon metal-ion binding. PMID- 15549762 TI - Molecular and electronic structure of four- and five-coordinate cobalt complexes containing two o-phenylenediamine- or two o-aminophenol-type ligands at various oxidation levels: an experimental, density functional, and correlated ab initio study. AB - The bidentate ligands N-phenyl-o-phenylenediamine, H(2)((2)L(N)IP), or its analogue 2-(2-trifluoromethyl)anilino-4,6-di-tert-butylphenol, ((4)L(N)IP), react with [Co(II)(CH(3)CO(2))(2)]4H(2)O and triethylamine in acetonitrile in the presence of air yielding the square-planar, four-coordinate species [Co((2)L(N))(2)] (1) and [Co((4)L(O))(2)] (4) with an S=1/2 ground state. The corresponding nickel complexes [Ni((4)L(O))(2)] (8) and its cobaltocene reduced form [Co(III)(Cp)(2)][Ni((4)L(O))(2)] (9) have also been synthesized. The five coordinate species [Co((2)L(N))(2)(tBu-py)] (2) (S=1/2) and its one-electron oxidized forms [Co((2)L(N))(2)(tBu-py)](O(2)CCH(3)) (2 a) or [Co((2)L(N))(2)I] (3) with diamagnetic ground states (S=0) have been prepared, as has the species [Co((4)L(O))(2)(CH(2)CN)] (7). The one-electron reduced form of 4, namely [Co(Cp)(2)][Co((4)L(O))(2)] (5) has been generated through the reduction of 4 with [Co(Cp)(2)]. Complexes 1, 2, 2 a, 3, 4, 5, 7, 8, and 9 have been characterized by X-ray crystallography (100 K). The ligands are non-innocent and may exist as catecholate-like dianions ((2)L(N)IP)(2-), ((4)L(N)IP)(2-) or pi radical semiquinonate monoanions ((2)L(N)ISQ)(*) (-), ((4)L(N)ISQ)(*) (-) or as neutral benzoquinones ((2) L(N)IBQ)(0), ((4) L(N)IBQ)(0); the spectroscopic oxidation states of the central metal ions vary accordingly. Electronic absorption, magnetic circular dichroism, and EPR spectroscopy, as well as variable temperature magnetic susceptibility measurements have been used to experimentally determine the electronic structures of these complexes. Density functional theoretical (DFT) and correlated ab initio calculation have been performed on the neutral and monoanionic species [Co((1)L(N))(2)](0,-) in order to understand the structural and spectroscopic properties of complexes. It is shown that the corresponding nickel complexes 8 and 9 contain a low-spin nickel(II) ion regardless of the oxidation level of the ligand, whereas for the corresponding cobalt complexes the situation is more complicated. Spectroscopic oxidation states describing a d(6) (Co(III)) or d(7) (Co(II)) electron configuration cannot be unambiguously assigned. PMID- 15549764 TI - Formation of organolithium hetero-aggregates [Li4Ar2(nBu)2] (Ar=C6H4CH(Me)NMe2-2) during the directed ortho-lithiation of [1-(dimethylamino)ethyl]benzene. AB - (R)-[1-(Dimethylamino)ethyl]benzene reacts with nBuLi in a 1:1 molar ratio in pentane to quantitatively yield a unique hetero-aggregate (2 a) containing the lithiated arene, unreacted nBuLi, and the complexed parent arene in a 1:1:1 ratio. As a model compound, [Li(4)(C(6)H(4)CH(Me)NMe(2)-2)(2)(nBu)(2)] (2 b) was prepared from the quantitative redistribution reaction of the parent lithiated arene Li(C(6)H(4)CH(Me)NMe(2)-2) with nBuLi in a 1:1 molar ratio. The mono-Et(2)O adduct [Li(4)(C(6)H(4)CH(Me)NMe(2)-2)(2)(nBu)(2)(OEt(2))] (2 c) and the bis Et(2)O adduct [Li(4)(C(6)H(4)CH(Me)NMe(2)-2)(2)(nBu)(2)(OEt(2))(2)] (2 d) were obtained by re-crystallization of 2 b from pentane/Et(2)O and pure Et(2)O, respectively. The single-crystal X-ray structure determinations of 2 b-d show that the overall structural motifs of all three derivatives are closely related. They are all tetranuclear Li aggregates in which the four Li atoms are arranged in an almost regular tetrahedron. These structures can be described as consisting of two linked dimeric units: one Li(2)Ar(2) dimer and a hypothetical Li(2)nBu(2) dimer. The stereochemical aspects of the chiral Li(2)Ar(2) fragment are discussed. The structures as observed in the solid state are apparently retained in solution as revealed by a combination of cryoscopy and (1)H, (13)C, and (6)Li NMR spectroscopy. PMID- 15549765 TI - "Bent bonds" between bismuth and carbon atoms as a result of C-H activation in Mo Bi complexes. AB - The reaction of molybdocenedihydride with two equivalents of [Bi(OtBu)(3)] proceeds via alcohol elimination and provides the compound [Cp(2)Mo{Bi(OtBu)(2)}(2)] (1), which contains two Mo--Bi metal bonds, in good yields. If the two reagents are employed in a 1:1 ratio continuative condensation reactions occur. These initially lead to [{Cp(2)Mo}(2){mu-Bi(OtBu)}(2)] (2), which, however, is very unstable in solution and decomposes via additional alcohol elimination: Complex-induced proximity effects facilitate the cleavage of C--H bonds within the cyclopentadienyl ligands by the residual alkoxide ligands, so that spontaneously two further equivalents of alcohol are released, thereby yielding two isomeric compounds 3 and 4 with Cp ligands bridging Mo--Bi metal bonds: The first isomer (3) contains two mu(2)-eta(5):eta(1)-C(5)H(4) ligands, the second isomer (4) contains one bridging mu(3)-eta(5):eta(1):eta(1)-C(5)H(3) ligand. The binding of these ligands to molybdenum and bismuth atoms at the same time is made possible through "bent bonds" between the bismuth and certain carbon centres. These unusual bonding situations were analysed by means of calculations based on density functional theory (DFT), the atoms in molecules (AIM) theory, natural bond order (NBO) considerations and the electron localisation function (ELF). According to the results the bonds can be understood in terms of carbanionic centres interacting with bismuth cations (i.e. closed-shell interactions). The formation of these bonds and the thermodynamics/kinetics involved on going from 2 to 3 and 4 were also studied by theoretical methods, so that the product formation is rationalised. The crystal structures of all four new compounds were determined. These structures but also the properties and mechanisms of formation are discussed against the background of the corresponding results obtained while studying the system [(Me)Cp(2)MoH(2)]/[Bi(OtBu)(3)]. PMID- 15549766 TI - Dimeric palladium complexes with bridging aryl groups: when are they stable? AB - Stable dimeric palladium(II) complexes of general formula [Pd(2)(mu-R)(2)(eta(3) allyl)(2)] (R=haloaryl, mesityl) have been prepared. Their X-ray crystal structures, determined for some of the complexes, show that the two coordination square planes are usually coplanar. The haloaryl complexes are fluxional in solution, showing exchange between cis and trans isomers (relative to the orientation of the two allyl groups in the dimer) through solvent-assisted associative bridge splitting. A number of other ancillary ligands (O,O, S,S, or C,N donors) failed to stabilize the bridging situation. Also, bridging phenyls were unstable. The reasons for this behavior and the formation of alternative compounds in attempts at synthesizing them are fully analyzed and explained. Stable aryl bridges seem to be favored by a combination of factors: the use of ancillary ligands of small size and lacking electron lone pairs, and the use of aryl ligands reluctant to homo and hetero C--C coupling. These seem to be more important factors in the stabilization of bridging aryl complexes than the strength of the bridges themselves. PMID- 15549767 TI - Dynamic equilibrium between a supramolecular capsule and bowl generated by inter- and intramolecular metal clipping. AB - The metal-induced self-assembly of a resorcin[4]arene derivative 1 that has four pyridine units as pendent groups and two equivalents of [M(dppp)(OTf)(2)] (M=Pd, Pt) results in a dynamic equilibrium between an interclipped supramolecular capsule 3 and an intraclipped bowl 4 in nitromethane, although the interclipped capsule 3 is formed as a sole adduct in chloroform/methanol and the intraclipped bowl 4 is formed exclusively in an aqueous phase. This demonstrates how metal induced self-assembly can be tuned by subtle changes in the solvent system. The coexistence of the two structures in nitromethane was characterized by NMR spectroscopy and coldspray ionization mass spectrometry (CSI-MS). The crystal structure of the interclipped capsule 3 b, which is composed of two units of ligand 1 and four Pt(II) ions, reveals the capsule cavity to have nanoscale dimensions of 15x20 A. NMR spectra show that the dynamic equilibrium between 3 and 4 is dependent on concentration and temperature. Temperature-dependent (1)H NMR spectroscopy was carried out from 273 to 343 K to verify the thermodynamic parameters that control the dynamic equilibrium process; the conversion from the interclipped supramolecular capsule 3 a to the intraclipped bowl 4 a is entropically favored and enthalpically disfavored. The rotational barrier of the restricted rotation of pyridine units in the intraclipped bowl 4 was determined by line-shape analysis. PMID- 15549769 TI - On the electronic structure of ethidium. AB - The electronic structure of the common intercalating agent ethidium bromide (3,8 diamino-5-ethyl-6-phenylphenanthridinium bromide) is dominated by an interplay of electron donating and withdrawing effects mediated by its nitrogen atoms. X-ray crystallography, UV/Vis and IR absorption, fluorescence emission, and NMR spectroscopy are used to probe the electronic properties of the phenanthridinium "core" of ethidium as well as its exocyclic amines and 6-phenyl groups. Interestingly, despite its positive charge, most of ethidium's aromatic carbon and hydrogen atoms have high electron densities (compared to both 6 phenylphenanthridine and benzene). The data suggest that electron donation by ethidium's exocyclic amines dominates over the electron withdrawing effects of its endocyclic iminium in their combined influence on the electron densities of these atoms. Ethidium's nitrogen atoms are, conversely, electron deficient where the 5-position is the most electropositive, followed by the 3-amino, and lastly the 8-amino group. These results have been used to generate an empirically-based pi-electron density map of ethidium that may prove useful to understanding its nucleic acid binding specificity. PMID- 15549770 TI - LIGHT costimulates CD40 triggering and induces immunoglobulin secretion; a novel key partner in T cell-dependent B cell terminal differentiation. AB - The T cell-dependent differentiation and function of B lymphocytes are tightly regulated by TNF ligands (L) and receptors interactions, such as CD40/CD40L, CD27/CD70 and CD134/CD314L. The LIGHT/HVEM system [homologous to lymphotoxin, inducible expression, competing for GpD of herpes virus, that binds to the herpes virus entry mediator (HVEM), and is expressed on activated T lymphocytes) focused our attention since HVEM has a large distribution that, in addition to T cells, DC or NK, includes tumor and normal B lymphocytes. HVEM was expressed on memory and naive B cells from peripheral blood or tonsils, but not on germinal center (GC) B cells. Costimulation by CD40L+LIGHT induced LIGHT expression at the B lymphocyte surface by a transcriptional mechanism since we detected de novo expression of LIGHT-specific mRNA. LIGHT expression was further enhanced by triggering of surface IgM, a stimulus that mimics a normal step of B cell physiology, i.e. specific antigen encounter. Stimulation by LIGHT increased the B cell proliferation induced by CD40L, and induced IgG and IgM (but not IgA) secretion. We conclude that LIGHT costimulation, that mimics the B cell encounter with activated LIGHT-expressing T lymphocytes, enhances both B cell proliferation and Ig production, and thus has a central importance for humoral immunity development. PMID- 15549772 TI - Preparation and multicolor electrochromic performance of a WO3/tris(2,2' bipyridine)ruthenium(II)/polymer hybrid film. AB - A tungsten trioxide (WO(3))/tris(2,2'-bipyridine)ruthenium(II) ([Ru(bpy)(3)](2+); bpy=2,2'-bipyridine)/poly(sodium 4-styrenesulfonate) (PSS) hybrid film was prepared by electrodeposition from a colloidal triad solution containing peroxotungstic acid (PTA), [Ru(bpy)(3)](2+), and PSS. A binary solution of [Ru(bpy)(3)](2+) and PTA (30 vol % ethanol in water) gradually gave an orange precipitate, possibly caused by the electrostatic interaction between the cationic [Ru(bpy)(3)](2+) and the anionic PTA. The addition of PSS to the binary PTA/[Ru(bpy)(3)](2+) solution remarkably suppressed this precipitation and caused a stable, colloidal triad solution to form. The spectrophotometric measurements and lifetime analyses of the photoluminescence from the excited [Ru(bpy)(3)](2+) ion in the colloidal triad solution suggested that the [Ru(bpy)(3)](2+) ion is partially shielded from electrostatic interaction with anionic PTA by the anionic PSS polymer chain. The formation of the colloidal triad made the ternary [Ru(bpy)(3)](2+)/PTA/PSS solution much more redox active. Consequently, the rate of electrodeposition of WO(3) from PTA increased appreciably by the formation of the colloidal triad, and fast electrodeposition is required for the unique preparation of this hybrid film. The absorption spectrum of the [Ru(bpy)(3)](2+) ion in the film was close to its spectrum in water, but the photoexcited state of the [Ru(bpy)(3)](2+) ion was found to be quenched completely by the presence of WO(3) in the hybrid film. The cyclic voltammogram (CV) of the hybrid film suggested that the [Ru(bpy)(3)](2+) ion performs as it is adsorbed onto WO(3) during the electrochemical oxidation. An ohmic contact between the [Ru(bpy)(3)](2+) ion and the WO(3) surface could allow the electrochemical reaction of adsorbed [Ru(bpy)(3)](2+). The composition of the hybrid film, analyzed by electron probe microanalysis (EPMA), suggested that the positive charge of the [Ru(bpy)(3)](2+) ion could be neutralized by partially reduced WO(3)(-) ions, in addition to Cl(-) and PSS units, based on the charge balance in the film. The electrostatic interaction between the WO(3)(-) ion and the [Ru(bpy)(3)](2+) ion might be responsible for forming the electron transfer channel that causes the complete quenching of the photoexcited [Ru(bpy)(3)](2+) ion, as well as the formation of the ohmic contact between the [Ru(bpy)(3)](2+) ion and WO(3). A multicolor electrochromic performance of the WO(3)/[Ru(bpy)(3)](2+)/PSS hybrid film was observed, in which transmittances at 459 and 800 nm could be changed, either individually or at once, by the selection of a potential switch. Fast responses, of within a few seconds, to these potential switches were exhibited by the electrochromic hybrid film. PMID- 15549771 TI - Down-regulation of CXCR4 expression on human CD8+ T cells during peripheral differentiation. AB - Multi-color flow cytometric analysis on human CD8(+) T cell subsets revealed that CXCR4 is predominantly expressed on CD8(+) T cells with the naive CD27(+)CD28(+)CD45RA(+) phenotype, and is down-regulated during differentiation into those with an effector phenotype. The down-regulation of CXCR4 expression during peripheral differentiation was supported by the fact that the expression of CXCR4 on CD8(+) T cells was negatively correlated with that of perforin. The analysis of CCR5, CCR7, and CXCR4 co-expression further showed that CD8(+) T cells expressing a high level of CXCR4 are CCR7(+)CCR5(-) naive or central memory subsets, and those expressing a low level of CXCR4 were included in the CCR7( )CCR5(+/-) memory/effector and effector subsets. Epstein Barr virus-specific CD8(+) T cells, which mostly express the memory phenotype, expressed CXCR4, while human cytomegalovirus-specific CD8(+) T cells, which mostly express the effector phenotype, partially expressed this receptor, showing that the expression of CXCR4 is also down-regulated during differentiation of viral antigen-specific CD8(+) T cells. The classification of human CD8(+) T cells based on the expression of these chemokine receptors should prove useful for studies that clarify the differentiation of human CD8(+) T cells. PMID- 15549773 TI - Effector and regulatory T cells derived from the same T cell clone differ in MHC class II-peptide multimer binding. AB - MHC class II-peptide multimers are a valuable tool for antigen-specific detection of CD4(+) T cells. However, it has been proposed that T cells in a hypo responsive state can have diminished binding of such multimers. In the present study, we investigated this phenomenon at the clonal level. We found that anergic CD4(+) T cells had a reduced capacity to bind MHC class II-peptide multimers compared to their non-anergic counterparts. Increasing the incubation temperature, time, or MHC-peptide valency could not equalize multimer binding by anergic and non-anergic T cells. Neither anergic T cells nor non-anergic T cells internalized the MHC class II-peptide dimers efficiently, and in both cases the dimers bound to the plasma membrane at locations containing a low amount of raft associated lipids. Disruption of lipid rafts, however, led to decreased dimer binding by non-anergic T cells and to a lesser extent by anergic T cells. Finally, we show that the depth of the anergic state of the T cell, which determines its ability to regulate other T cell responses, correlates with the reduced dimer binding. We here demonstrate for the first time differential MHC class II-peptide multimer binding by regulatory (anergic) and effector T cells with identical TCR. PMID- 15549774 TI - Development of CD1d-restricted NKT cells in the mouse thymus. AB - Using genetic and phenotypic analyses, we have analyzed the developmental pathway of mouse CD1d-restricted invariant NKT cells. We provide strong evidence that similar to conventional T cells, positive selection of NKT cells occurs during a CD4(+)CD8(+) stage. Later stages of NKT cell development involved the down regulation of both TCR and CD4 levels and therefore diverge from conventional T cell development pathways. A unique and complete dependency for development on Fyn, a Src family kinase member, also distinguishes the NKT cell and conventional T cell populations. PMID- 15549775 TI - Inhibition of altered peptide ligand-mediated antagonism of human GAD65 responsive CD4+ T cells by non-antagonizable T cells. AB - Altered peptide ligands derived from T cell-reactive self antigens have been shown to be protective therapeutic agents in animal models of autoimmunity. In this study we identified several altered peptide ligands derived from the type 1 diabetes-associated autoantigen human glutamic acid decarboxylase 65 (hGAD65) epitope that were capable of antagonizing a subset of a panel of human CD4(+) GAD65 (555-567)-responsive T cell clones derived from a diabetic individual. While no altered peptide ligand was able to antagonize all six clones in the T cell panel, a single-substituted peptide of isoleucine to methionine at position 561, which resides at the TCR contact p5 position, was able to antagonize five out of the six hGAD65-responsive clones. In a mixed T cell culture system we observed that altered peptide ligand-mediated antagonism is inhibited in a dose dependent manner by the presence of non-antagonizable hGAD65 (555-567)-responsive T cells. From an analysis of the cytokines present in the mixed T cell cultures, interleukin-2 was sufficient to inhibit altered peptide ligand-induced antagonism. The inhibition of altered peptide ligand-mediated antagonism of self antigen-responsive T cells by non-antagonizable T cells has implications in altered peptide ligand therapy where T cell antagonism is the goal. PMID- 15549776 TI - B cells alter the phenotype and function of follicular-homing CXCR5+ T cells. AB - The CXC chemokine receptor (CXCR)5 is rapidly induced on activated CD4(+) T cells, allowing migration toward secondary lymphoid tissue follicles, where the CXCR5 ligand CXCL13/BCA-1 is produced. Such CXCR5(+) T cells provide efficient help for B cell immunoglobulin production and are termed follicular B helper T (T(FH)) cells. However, the molecular mechanisms by which T(FH) cells provide B cell help are unknown. Here, we demonstrate that newly generated (antigen-primed) T(FH) cells express a phenotype consistent with induction of B cell proliferation, but co-culture with primed B cells resulted in a switch to a plasma cell-inducing phenotype, characterized by loss of CD154, induction of CD70 and an increase in IL-10 production capacity. The ability to produce IL-10 could be maintained as a stable phenotype, but its secretion was strictly dependent on inducible costimulator (ICOS) signaling. Furthermore, B cells preserved a lymph node migration phenotype in proliferating T(FH) cells by preventing the loss of CC chemokine receptor (CCR)7 and the induction of CCR5. Thus, B cells directly modulate the B cell helper phenotype in T(FH) cells and actively promote their prolonged co-localization with these cells. PMID- 15549777 TI - Frontline: Inhibition of allergen-induced pulmonary inflammation by the tripeptide feG: a mimetic of a neuro-endocrine pathway. AB - Interactions between the neuro-endocrine system and immune system help maintain health. One interaction involves the superior cervical ganglia (SCG), which regulate the prohormone submandibular rat 1 (SMR1) produced by the submandibular gland (SMG). A peptide derived from SMR1, feG, has anti-inflammatory activity, and modification to D-isomer feG enhances bioactivity. We tested feG as a therapeutic agent for airways inflammation, using rats sensitized by OVA or Nippostrongylus brasiliensis (Nb). Treatment with feG but not fdG down-regulated OVA-challenge-induced increases in bronchoalveolar lavage (BAL)-derived macrophages, eosinophils and PMN (neutrophils) by 44%, 69% and 67%, respectively, at 24 h. We found that feG also reduced ICAM-1 on BAL-derived macrophages and eosinophils by 27% and 65%, and L-selectin on PMN by 55% following OVA challenge. Furthermore, feG but not fdG reduced the OVA-induced TNF increase in BAL fluid. We showed that feG also down-regulated both hyper-responsiveness to methacholine (by 27%) and microgranulomata formation in the lung parenchyma. In Nb-challenged rats, feG treatment inhibited ex vivo allergen-induced contraction of tracheal smooth muscle by up to 73%. In conclusion, feG, which is a mimetic of a peptide derived from a rat salivary gland prohormone, has anti-inflammatory properties in allergic airways inflammation in Brown-Norway rats. The role of the SCG-SMG neuro endocrine pathway in allergic asthma and other inflammatory diseases requires additional study. PMID- 15549778 TI - Bile duct cysts in adults. AB - BACKGROUND: Bile duct cysts are rare and of uncertain origin. Most have been reported in young females of Asian descent, but an increasing number have occurred in Western adults. METHODS: A Medline literature search was performed to locate articles on the pathophysiological concepts, clinical behaviour and management controversies pertaining to bile duct cysts in adults. Emphasis was placed on reports from the past two decades. RESULTS AND CONCLUSION: An increasing rate of occurrence of bile duct cyst is reported in adults. Type IV cysts are more frequent in adults than children. Presentation tends to be non specific abdominal discomfort. Related hepatobiliary or pancreatic disease frequently precedes recognition, and may complicate the postoperative course. Surgical treatment aims to relieve complications deriving from the cysts and to reduce the significant risk of malignant change within the biliary tree. Complete cyst resection, cholecystectomy and Roux-en-Y hepaticojejunostomy reconstruction is standard. Controversy exists about the role of hepatic resection in type IV and V cysts, and the role of minimally invasive and laparoscopic treatment. In general, the outcome is good and a near-zero mortality rate has been reported in institutional series over the past decade. PMID- 15549780 TI - The governance of innovation. PMID- 15549779 TI - Intraoperative adverse events and outcome after anterior resection of the rectum. AB - BACKGROUND: The aim of this population-based study was to analyse the relationship between intraoperative adverse events and outcome after anterior resection. METHODS: All 140 patients who underwent elective anterior resection in Sweden between 1987 and 1995, and who died within 30 days, were compared with a group of 423 randomly selected patients who underwent the same procedure during the same interval but survived the operation. Intraoperative adverse events and intraoperative measures taken were analysed in relation to outcome of surgery. RESULTS: Of those who died, 45.7 per cent had intraoperative adverse events compared with 30.3 per cent in the cohort group. Major bleeding, gross spillage of faeces, and two or more intraoperative adverse events were more common among those who died. When the anastomosis was considered unsatisfactory, it was more frequently reconstructed (restapled or completely resutured), with or without a temporary stoma, in those who survived. The use of a temporary stoma was comparable in the two groups when adverse events were present. CONCLUSION: Intraoperative adverse events were important contributors to morbidity and mortality. Complete reconstruction of an unsatisfactory anastomosis, with or without addition of a temporary stoma, was more frequently performed in the survivors, and may have diminished the risk of postoperative death. PMID- 15549781 TI - In vitro and in vivo biocompatibility evaluation of a polyalkylimide hydrogel for soft tissue augmentation. AB - Injectable fillers are commonly used in Plastic and Reconstructive Surgery to correct serious and slight aesthetic defects due to their low invasiveness and an easy implant technique procedure. Synthetic hydrogels are proposed as filler materials for their similarity with soft tissue and to avoid many disadvantages of naturally derived materials such as short persistence, allergenicity, and immunogenicity. Our studies are focused on the biocompatibility evaluation of a polyacrylic hydrogel containing alkylimide-amide groups and pyrogen free water (96%) (Bio-Alcamid by means of the in vitro cytotoxicity and mutagenicity assays and the in vivo skin irritation, sensitization test, and subcutaneous implant. All tests conducted on Bio-Alcamid showed no toxicity. It is a substance easy to inject and remove; it does not migrate, and its safety allows it to be a suitable filler for the correction of slight and also very serious aesthetic defects. PMID- 15549782 TI - Human skin cell cultures onto PLA50 (PDLLA) bioresorbable polymers: influence of chemical and morphological surface modifications. AB - Poly(alpha-hydroxy acid)s derived from lactic and glycolic acid are bioresorbable polymers which can cover a large range of thermal, physical, mechanical, and biological properties. Human keratinocytes have been shown as able to grow on a poly(DL-lactic acid) film. However the keratinocyte growth was delayed with respect to culture on standard tissue culture polystyrene, even though the same plateau level was observed after 2 weeks. In order to improve the performance of poly(DL-lactic acid) films as skin culture support, their surface was modified by creating tiny cavities using a method based on the leaching out of poly(ethylene oxide) from poly(lactic acid)-poly(ethylene oxide) heterogeneous blends. The surface of the films was also chemically modified by alkaline attack with sodium hydroxide and by type-I collagen coating. Murine fibroblast cell line and primary cultures of human fibroblasts and of two types of keratinocytes were allowed to adhere and to grow comparatively on the different films. The presence of cavities affected neither the adhesion of dermal fibroblasts nor that of keratinocytes. Only keratinocyte proliferation was significantly reduced by the presence of cavities. Collagen coating improved skin cell adhesion and proliferation as well, except in the case of murine fibroblasts. In the case of the NaOH treatments, similar trends were observed but their extent depended on the treatment time. In the case of chemical modifications, fluorescence microscopy bore out adhesion and proliferation tendencies deduced from MTT tests. PMID- 15549783 TI - Hydroxyapatite and gelatin composite foams processed via novel freeze-drying and crosslinking for use as temporary hard tissue scaffolds. AB - Hydroxyapatite (HA) and gelatin composites were fabricated in a foam type via a novel freeze-drying and crosslinking technique. The morphological and mechanical properties of and in vitro cellular responses to the foams were investigated. The HA powder was added at up to 30 wt % into the gelatin solution, and the mixtures were freeze-dried and further crosslinked. The pure gelatin foam had a well developed pore configuration with porosity and pore size of approximately 90% and 400-500 microm, respectively. With HA addition, the porosity decreased and pore shape became more irregular. The HA particulates, in sizes of about 2-5 microm, were distributed within the gelatin network homogeneously and made the framework surface rougher. All the foams had high water absorption capacities, showing typical hydrogel characteristics, even though the HA addition decreased the degree of water absorption. The HA addition made the foam much stronger and stiffer (i.e., with increasing HA amount the foams sustained higher compressive stress and had higher elastic modulus in both dry and wet states). The osteoblast like human osteosarcoma cells spread and grew actively on all the foams. The cell proliferation rate, quantified indirectly on the cells cultured on Ti discs coated with gelatin and gelatin-HA composites using MTT assay, exhibited an up regulation with gelatin coating compared with bare Ti substrate, but a slight decrease on the composite coatings. However, the alkaline phosphatase activities expressed by the cells cultured on composites foams as well as their coatings on Ti discs were significantly enhanced compared with those on pure gelatin foam and coating. These findings suggest that the gelatin-HA composite foams have great potential for use as hard tissue regeneration scaffolds. PMID- 15549784 TI - Photochemistry and photophysics of halogen-substituted dibenzothiophene oxides. AB - Dibenzothiophene-5-oxide (DBTO) cleanly produces dibenzothiophene (DBT) on direct photolysis, but with very low quantum yield. A proposed mechanism involves scission of the S-O bond which is coupled to an intersystem crossing step, thus producing the sulfide and O((3)P) via a unimolecular pathway. To test this hypothesis, heavy atom substituted DBTOs were prepared and photolyzed. Iodo-, bromo-, and chloro-substituted DBTOs show higher quantum yields for deoxygenation than does the parent molecule, in the order consistent with an intersystem crossing-related heavy atom effect. 2-Iododibenzothiophene also undergoes photochemical deiodination. Phosphorescence data are consistent with heavy-atom assisted intersystem crossing. PMID- 15549785 TI - Design, synthesis, and evaluation of a biomimetic artificial photolyase model. AB - Two new artificial photolyase models that recognize pyrimidine dimers in protic and aprotic organic solvents as well as in water through a combination of charge and hydrogen-bonding interactions and use a mimic of the flavine to achieve repair through reductive photoinduced electron transfer are presented. Fluorescence and NMR titration studies show that it forms a 1:1 complex with pyrimidine dimers with binding constants of approximately 10(3) M(-1) in acetonitrile or methanol, while binding constants in water at pH 7.2 are slightly lower. Excitation of the complex with visible light leads to clean and rapid cycloreversion of the pyrimidine dimer through photoinduced electron transfer catalysis. The reaction in water is significantly faster than in organic solvents. The reaction slows down at higher conversions due to product inhibition. PMID- 15549786 TI - Synthesis and activity of fluorescent isoprenoid pyrophosphate analogues. AB - New fluorescent analogues of farnesol and geranylgeraniol have been prepared and then converted to the corresponding pyrophosphates. These analogues incorporate anthranylate or dansyl-like groups anchored to the terpenoid skeleton through amine bonds that would be expected to be relatively stable to metabolism. After addition of the alcohols or the pyrophosphates to the culture medium, their fluorescence is readily observed inside a human-derived leukemia cell line. Enzyme assays have revealed that the farnesyl pyrophosphate analogue is an inhibitor of FTase, while the corresponding alcohol is not. These results, together with Western blot analyses of cell lysates, indicate that the farnesyl pyrophosphate analogue penetrates the cells as an intact pyrophosphate and that it does so at a biologically relevant concentration. PMID- 15549787 TI - Oligomers of hexa-peri-hexabenzocoronenes as "super-oligophenylenes": synthesis, electronic properties, and self-assembly. AB - Hexa-peri-hexabenzocoronene (HBC) is a remarkable polycyclic aromatic hydrocarbon and is often called "superbenzene" because of its similarity to benzene. In this article we present the facile synthesis of oligomers of HBC, up to trimers (3, 4, 5a-c) with different modes of connection. UV-vis and fluorescence spectroscopy studies reveal that the oligomers are electronically decoupled. This arises from the small atomic orbital coefficients of the bridge-head carbon atoms, the large torsion angle between the HBC units, and the large distance of interacting transition dipoles due to the size of the HBC chromophore. For comparison, a methylene-bridged HBC dimer 6, so-called "superfluorene", was prepared. The induced planarity improves pi-conjugation and suppresses the geometrical relaxation of the backbone upon electronic excitation, leading to a prominent 0-0 transition band in the fluorescence spectra. The self-assembly of the oligomers and of superfluorene 6 was studied by wide-angle X-ray diffraction (WAXD) in the bulk state, and ordered columnar stacking occurs in the HBC dimer 3, p-HBC trimer 4, and superfluorene 6. Measurements of shear-aligned samples show that, despite increasing aspects ratio by linear entrainment of disks, the anitropic element that is subject to alignment by shear is the supramolecular columns. PMID- 15549788 TI - Synthesis, basicity, and dynamics of a perfluorocyclohexenyl anion. AB - Electron transfer to perfluoro-1,3-dimethylcyclohexane in moist THF has yielded two quite different products. Tetrabutylammonium iodide irradiated with ultraviolet light gives a tetrabutylammonium enolate, but potassium fluorenone ketyl affords a cyclohexenyl anion. This allylic anion was isolated as its conjugate acid, a rather strong carbon acid. Ring inversion in the anion, measured by (19)F NMR line shape analysis, is characterized by these activation parameter values: DeltaH(++) = 8.84 +/-0.14 kcal/mol and DeltaS(++) = 0.81 +/- 0.6 cal mol(-1) K(-1). PMID- 15549790 TI - Aminolysis of aryl chlorothionoformates with anilines in acetonitrile: effects of amine nature and solvent on the mechanism. AB - The aminolysis of aryl chlorothionoformates (7, YC(6)H(4)OC(=S)Cl) with anilines (XC(6)H(4)NH(2)) in acetonitrile at 5.0 degrees C has been investigated. The rates are slower than those for the corresponding reactions of aryl chloroformates (6, YC(6)H(4)OC(=O)Cl). This rate sequence is a reverse of that for alkyl chloroformates (1-4) in water, for which rate-limiting formation of a tetrahedral intermediate, T(+/-), is predicted. On the basis of the large negative cross-interaction constant, rho(XY) = -0.77, failure of the reactivity selectivity principle, normal k(H)/k(D) values involving deuterated nucleophiles (XC(6)H(4)ND(2)), and low DeltaH(not equal) with large negative DeltaS(not equal) values, a concerted mechanism with a four-membered hydrogen bonded cyclic transition state (11) is proposed for the title reaction series. It has been shown that the solvent change from water to acetonitrile for the aminolysis of 6 and 7 causes a mechanistic change from stepwise to concerted. PMID- 15549789 TI - Electrophilic-induced cyclization reaction of hexahydroindolinone derivatives and its application toward the synthesis of (+/-)-erysotramidine. AB - A convenient synthesis of variously substituted octahydroindolo[7a,1a] isoquinolinones has been achieved by an acid-induced cyclization of hexahydroindolinones bearing tethered phenethyl groups. The formation of a single lactam diastereomer is the result of the stereoelectronic preference for axial attack by the aromatic ring onto the initially formed N-acyliminium ion from the least hindered side. Additional experiments showed that a variety of hexahydroindolinones containing tethered pi-bonds undergo a related acid-induced cyclization reaction. Treatment of the 3-methylbut-3-enyl-substituted hexahydroindolinone with acid furnished a 3:1 mixture of isomeric octahydropyrido[2,1-i]indolinones in near-quantitative yield. Interestingly, cyclization of the closely related 1-(3-methoxybut-3-enyl)-substituted hexahydroindolin-one afforded a pyrrolo[3,2,1-ij]quinolinone as the exclusive product. With this system, initial protonation takes place on the more nucleophilic enol ether pi-bond and the resulting carbonium ion undergoes a subsequent cyclization with the enamido pi-bond to give the observed product. The electrophilic promoted cyclizations were extended to include the related hexahydro[1]pyrindinone and 1H-quinolinone systems. An NBS-promoted intramolecular electrophilic aromatic substitution reaction of 1-[2-(3,4 dimethoxyphenyl)ethyl]-1,4,5,6-tetrahydroindolinone was used to assemble the tetracyclic core of the erythrinone skeleton. The resulting cyclized product was transformed into (+/-)-erysotramidine in three additional steps. PMID- 15549791 TI - Reverse micellar aggregates: effect on ketone reduction. 1. Substrate role. AB - The reduction of three aromatic ketones, acetophenone (AF), 4-methoxyacetophenone (MAF), and 3-chloroacetophenone (CAF), by NaBH(4) was followed by UV-vis spectroscopy in reverse micellar systems of water/AOT/isooctane at 25.0 degrees C (AOT is sodium 1,4-bis-2-ethylhexylsulfosuccinate). The first-order rate constants, k(obs), increase with the concentration of surfactant due to the substrate incorporation at the reverse micelle interface, where the reaction occurs. For all the ketones the reactivity is lower at the micellar interface than in water, probably reflecting the low affinity of the anionic interface for BH(4)(-). Kinetic profiles upon water addition show maxima in k(obs) at W(0) approximately 5 probably reflecting a strong interaction between water and the ionic headgroup of AOT; at W(0) < 5 by increasing W(0) BH(4)(-) is repelled from the anionic interface once the water pool forms. The order of reactivity was CAF >> AF > MAF. Application of a kinetic model based on the pseudophase formalism, which considers distribution of the ketones between the continuous medium and the interface, and assumes that reaction take place only at the interface, gives values of the rate constants at the interface of the reverse micellar system. At W(0) = 5, we conclude that NaBH(4) is wholly at the interface, and at W(0) = 10 and 15, where there are free water molecules, the partitioning between the interface and the water pool has to be considered. The results were used to estimate the ketone and borohydride distribution constants between the different pseudophases as well as the second-order reaction rate constant at the micellar interface. PMID- 15549792 TI - Reverse micellar aggregates: effect on ketone reduction. 2. Surfactant role. AB - Kinetics of the reduction of 3-chloroacetophenone (CAF) with sodium borohydride (NaBH(4)) were followed by UV-vis spectroscopy at 27.0 degrees C in different reverse micellar media, toluene/BHDC/water and toluene/AOT/water, and compared with results in an isooctane/AOT/water reverse micellar system. AOT is sodium 1,4 bis-2-ethylhexylsulfosuccinate, and BHDC is benzyl-n-hexadecyl dimethylammonium chloride. The kinetic profiles were investigated as a function of variables such as surfactant and NaBH(4) concentration and the amount of water dispersed in the reverse micelles, W(0) = [H(2)O]/[surfactant]. In all cases, the first-order rate constant, k(obs), increases with the concentration of surfactant as a consequence of incorporating the substrate into the interface of the reverse micelles where the reaction takes place. The reaction is faster at the cationic interface than at the anionic one probably because the negative ion BH(4)(-) is part of the cationic interface. The effect of the external solvent on the reaction shows that reduction is favored in the isooctane/AOT/water reverse micellar system than that with an aromatic solvent. This is probably due to BH(4)(-) being more in the water pool of the toluene/AOT/water reverse micellar system. The kinetic profile upon water addition depends largely on the type of interface. In the BHDC system, k(obs) increases with W(0) in the whole range studied while in AOT the kinetic profile has a maximum at W(0) approximately 5, probably reflecting the fact that BH(4)(-) is part of the cationic interface while, in the anionic one, there is a strong interaction between water and the polar headgroup of AOT below W(0) = 5 and, above that, BH(4)(-) is repelled from the interface once the water pool has formed. Application of a kinetic model based on the pseudophase formalism, which considers the distribution of the ketone between the continuous medium and the interface and assumes that reaction takes place only at the interface, has enabled us to estimate rate constants at the interface of the reverse micellar systems. At W(0) < 10, it was considered that NaBH(4) is wholly at the interface and, at W(0) >/= 10, where there are free water molecules, also the partitioning between the interface and the water pool was taken into account. The results were used to evaluate CAF and NaBH(4) distribution constants between the different pseudophases as well as the second-order reaction rate constant of the reduction reaction in the micellar interface. PMID- 15549793 TI - Challenging the auxiliary donor effect on molecular hyperpolarizability in thiophene-containing nonlinear chromophores: X-ray crystallographic and optical measurements on two new isomeric chromophores. AB - To reexamine the established "auxiliary donor" effect of thiophene in nonlinear optical (NLO) chromophores, we have prepared two isomeric donor-acceptor azo dyes, differing only in the position of the thiophene. Experimental analysis of these chromophores, including electric field-induced second harmonic generation (EFISH) and X-ray crystallography, contradicts previous experimental findings on similar chromophores but is consistent with the majority of computational precedents. We have found that the thiophene on the donor side produces a compound with a larger dipole moment; however, the isomer with the thiophene on the acceptor side is more nonlinear and has a higher figure of merit for NLO device applications. PMID- 15549794 TI - High kinetic resolution in the addition of a racemic allenylzinc onto enantiopure N-tert-butanesulfinimines: concise synthesis of enantiopure trans-2 ethynylaziridines. AB - Enantiopure trans-ethynyl N-tert-butanesulfinylaziridines (R(S))-6 were prepared in good to excellent yields by the condensation of the racemic allenylzinc species 1 derived from 3-chloro-1-trimethylsilylpropyne onto the corresponding enantiopure N-tert-butanesulfinimines (R(S))-5. The absolute stereochemistry of enantiopure N-tert-butanesulfinylaziridines (R(S))-6 was shown to be (R(S),2R,3R) and results from a chelate-type transition state in which the zinc atom of allenylzinc 1 is coordinated by both the nitogen and the oxygen atoms of the imine. Further removal of the N-tert-butanesulfinyl auxiliary of alkyl 3 substituted and 3,3-disubstituted ethynyl N-tert-butanesulfinylaziridines (R(S)) 6 could be achieved by treatment with HCl in MeOH affording the corresponding deprotected aziridines (2R,3R)-9 and (2R)-9 respectively as enantiomerically pure compounds. PMID- 15549795 TI - Synthesis of fused polycycles by 1,4-palladium migration chemistry. AB - Novel palladium migration/arylation methodology for the synthesis of complex fused polycycles has been developed, in which one or more sequential Pd-catalyzed intramolecular migration processes involving C-H activation are employed. The chemistry works best with electron-rich aromatics, which is in agreement with the idea that these palladium-catalyzed C-H activation reactions parallel electrophilic aromatic substitution. PMID- 15549796 TI - A facile regiocontrol in the palladium-catalyzed annulation of fluorine containing internal alkynes with variously substituted 2-iodoanilines: a new regioselective synthesis of 2- or 3-fluoroalkylated indole derivatives. AB - Treatment of various types of fluoroalkylated alkynes with o-iodoaniline in the presence of Pd(PPh(3))(4) in DMF at 80 degrees C for 8 h mainly gave 2 fluoroalkylated indoles in high yields. The use of P(o-Tol)(3) instead of PPh(3) as a ligand led to the preferential formation of 3-fluoroalkylated indoles in high yields. Interestingly, the reaction of trifluoromethylated alkynes bearing a benzylic substituent afforded 2- or 3-trifluoroethylated indole derivatives in good yields. PMID- 15549797 TI - Regio- and diastereoselective insertion of allenes into stable oxapalladacycles with a metal-bonded stereogenic carbon. preparation of contiguously substituted 3,4-dihydro-2H-1-benzopyrans. AB - Insertion of monosubstituted allenes into stable oxapalladacycle I was studied. The aim of this work was to define steric and electronic parameters of allenes that would allow for a regio- and diastereoselective synthesis of 2,3 disubstituted 3,4-dihydro-2H-1-benzopyrans, which could not be prepared via related catalytic protocols. Allenes with electron-donating alkyl substituents R sterically unencumbered at the C-3 and C-4 carbons reacted with palladacycles I to afford benzopyrans IV in good yields (45-81%), exclusively as cis diastereomers. Less than 10% of the regioisomeric benzopyrans V was detected in the crude reaction mixtures. Methoxy 1,2-propadiene afforded benzopyran IV in 98% yield as the trans diastereomer in 92% de. In contrast, allenes with electron withdrawing substituents yielded benzopyrans V with an E double bond exclusively. Nonracemic palladacycles featuring a palladium-bonded stereogenic carbon as the only element of asymmetry underwent the allene insertion with 63-93% retention of the stereochemical information, providing benzopyrans IV or V in 40-47% ee. These results demonstrated that O-bonded palladium enolates did not operate as predominant intermediates in the insertion process. The study highlights the configurational stability of carbon-bonded palladium ester enolates, especially notable in systems lacking chiral nonracemic auxiliary ligands. PMID- 15549798 TI - Synthesis of 1-amino-3-[(dihydroxyboryl)methyl]- cyclobutanecarboxylic acid as a potential therapy agent. AB - A novel boronated aminocyclobutanecarboxylic acid (1) was synthesized for potential use in boron neutron capture therapy. Starting from the readily available 3-(bromomethyl)cyclobutanone ketal (4), several synthetic routes to 1 were evaluated. After several unsuccessful attempts with traditional synthetic methods, a novel synthetic strategy to generate the new boronated cyclic amino acid was developed. The tolerance of the hydantoin group to the selenoxide elimination reaction conditions in the preparation of alkenyl compound 7 proved to be the key step in the new strategy. PMID- 15549799 TI - The RuO4-catalyzed ketohydroxylation. Part 1. Development, scope, and limitation. AB - A new straightforward oxidation of C,C-double bonds to unsymmetrical alpha hydroxy ketones using catalytic amounts of RuCl(3) and stoichiometric amounts of Oxone under buffered conditions has been developed, a reaction for which we coined the expression "ketohydroxylation". The transformation allows the direct formation of alpha-hydroxy ketones (acyloins) from olefins without intermediate formation of syn-diols. The present paper will provide detailed information starting with the underlying concept and the subsequent development of the reaction. The effect of base, solvent stoichiometry, and temperature will be discussed resulting in an improved mechanistic model that might help to explain the influence of different reaction parameters on reactivity and selectivity in RuO(4)-catalyzed oxidations of C,C-double bonds. Furthermore, an improved workup procedure allows the recovery of the ruthenium catalyst by precipitation while simplifying the overall product purification. The second part of the paper focuses on exploration of scope and limitation. A variety of substituted olefins are oxidized to alpha-hydroxy ketones in good to excellent regioselectivities and yield. Cyclic substrates proved to be problematic to oxidize; however, a careful analysis of temperature effects resulted in the development of a successful protocol for the ketohydroxylation of cyclic substrates by simply decreasing the reaction temperature. PMID- 15549800 TI - A novel redox reaction between 8-Aza-5,7-dimethyl-2-trifluoromethylchromone and alkyl mercaptoacetates. Facile synthesis of CF3-containing 2-pyridone derivatives. AB - 8-Aza-5,7-dimethyl-2-trifluoromethylchromone reacts with alkyl mercaptoacetates in the presence of triethylamine to give pyrido derivatives of 2-oxa-7 thiabicyclo[3.2.1]octane, which undergo the reductive ring opening to alkyl 2-[[3 (4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)-3-oxo-1 (trifluoromethyl)propyl]sulfanyl]acetates. The latter can be also obtained directly from 8-aza-5,7-dimethyl-2-trifluoromethylchromone and behave as the masked alpha,beta-unsaturated ketone, 4,6-dimethyl-3-(4,4,4-trifluorobut-2 enoyl)pyridin-2(1H)-one. This compound was independently synthesized from 3 acetyl-4,6-dimethyl-2-pyridone, and its synthetic potential was studied. A wide variety of 2-pyridone derivatives containing the CF(3) group have been prepared in good to moderate yields. PMID- 15549801 TI - Improved synthesis of aryltrialkoxysilanes via treatment of aryl Grignard or lithium reagents with tetraalkyl orthosilicates. AB - General reaction conditions for the synthesis of aryl(trialkoxy)silanes from aryl Grignard and lithium reagents and tetraalkyl orthosilicates (Si(OR)(4)) have been developed. Ortho-, meta-, and para-substituted bromoarenes underwent efficient metalation and silylation at low temperature to provide aryl siloxanes. Mixed results were obtained with heteroaromatic substrates: 3-bromothiophene, 3-bromo-4 methoxypyridine, 5-bromoindole, and N-methyl-5-bromoindole underwent silylation in good yield, whereas a low yield of siloxane was obtained from 2-bromofuran, and 2-bromopyridine failed to give silylated product. The synthesis of siloxanes via organolithium and magnesium reagents was limited by the formation of di- and triarylated silanes (Ar(2)Si(OR)(2) and Ar(3)SiOR, respectively) and dehalogenated (Ar-H) byproducts. Silylation at low temperature gave predominantly monoaryl siloxanes, without requiring a large excess of the electrophile. Optimal reaction conditions for the synthesis of siloxanes from aryl Grignard reagents entailed addition of arylmagnesium reagents to 3 equiv of tetraethyl- or tetramethyl orthosilicate at -30 degrees C in THF. Aryllithium species were silylated using 1.5 equiv of tetraethyl- or tetramethyl orthosilicate at -78 degrees C in ether. PMID- 15549802 TI - Photochemical acetalization of carbonyl compounds in protic media using an in situ generated photocatalyst. AB - Carbonyl compounds are conveniently converted into their corresponding dimethyl acetals in good yields and short reaction times by means of a photochemical reaction in methanol with a catalytic amount of chloranil (2,3,5,6-tetrachloro 1,4-benzoquinone, CA) as the sensitizer. Using aldehydes gives better results than using ketones, which also tend to form enol ethers as side products. These results are similar to those of simple acid-catalyzed acetalization reactions, suggesting the involvement of a photochemically generated acid. On the basis of steady state and laser flash photolysis data the reaction is proposed to involve the in situ generation of a photocatalyst (2,3,5,6-tetrachloro-1,4-hydroquinone, TCHQ) via reaction of CA with the solvent. The acetalization process is initiated by ionization of TCHQ, followed by loss of a proton to the solvent or the carbonyl, which starts a catalytic reaction. The photocatalyst is regenerated via a disproportionation reaction. PMID- 15549803 TI - Steady-state and laser flash photolysis study of the carbon-carbon bond fragmentation reactions of 2-arylsulfanyl alcohol radical cations. AB - The N-methylquinolinium tetrafluoroborate (NMQ(+))-sensitized photolysis of the erythro-1,2-diphenyl-2-arylsulfanylethanols 1-3 (1, aryl = phenyl; 2, aryl = 4 methylphenyl; 3, aryl = 3-chlorophenyl) has been investigated in MeCN, under laser flash and steady-state photolysis. Under laser irradiation, the formation of sulfide radical cations of 1-3, in the monomeric (lambda(max) = 520-540 nm) and dimeric form (lambda(max) = 720-->800 nm), was observed within the laser pulse. The radical cations decayed by first-order kinetics, and under nitrogen, the formation of ArSCH(*)Ph (lambda(max) = 350-360 nm) was clearly observed. This indicates that the decay of the radical cation is due to a fragmentation process involving the heterolytic C-C bond cleavage, a conclusion fully confirmed by steady-state photolysis experiments (formation of benzaldehyde and the dimer of the alpha-arylsulfanyl carbon radical). Whereas the fragmentation rate decreases as the C-C bond dissociation energy (BDE) increases, no rate change was observed by the replacement of OH by OD in the sulfide radical cation (k(OH)/k(OD) = 1). This suggests a transition state structure with partial C-C bond cleavage where the main effect of the OH group is the stabilization of the transition state by hydrogen bonding with the solvent. The fragmentation rate of 2-hydroxy sulfanyl radical cations turned out to be significantly slower than that of nitrogen analogues of comparable reduction potential, probably due to a more efficient overlap between the SOMO in the heteroatom and the C-C bond sigma-orbital in the second case. The fragmentation rates of 1(+*)-3(+*) were found to increase by addition of a pyridine, and plots of k(base) against base strength were linear, allowing calculation of the beta Bronsted values, which were found to increase as the reduction potential of the radical cation decreases, beta = 0.21 (3(+*)), 0.34 (1(+*)), and 0.48 (2(+*)). The reactions of 1(+*) exhibit a deuterium kinetic isotope effect with values that increase as the base strength increases: k(OH)/k(OD) = 1.3 (pyridine), 1.9 (4-ethylpyridine), and 2.3 (4-methoxypyridine). This finding and the observation that with the above three bases the rate decreases in the order 3(+*) > 1(+*) > 2(+*), i.e., as the C-C BDE increases, suggest that C-C and O-H bond cleavages are concerted but not synchronous, with the role of OH bond breaking increasing as the base becomes stronger (variable transition state). It is probable that, with the much stronger base, 4 (dimethylamino)pyridine, a change to a stepwise mechanism may occur where the slow step is the formation of a radical zwitterion that then rapidly fragmentates to products. PMID- 15549804 TI - Synthesis and application of carbohydrate-derived morpholine amino acids. AB - The synthesis of series of diversely functionalized epsilon-morpholine amino acids (MAAs, 5a-h) starting from an epsilon-sugar amino acid and following a two step oxidative glycol cleavage/reductive amination strategy, is described. In an alternative synthetic scheme, diastereoisomerically pure delta-MAAs (12a,b) were obtained. Oligopeptides containing MAAs were prepared either by direct incorporation of a MAA building block or by subjecting a fully assembled SAA containing peptide to the two-step glycol cleavage/reductive amination procedure. PMID- 15549805 TI - High-load, soluble oligomeric carbodiimide: synthesis and application in coupling reactions. AB - A facile preparation of a high-load, soluble oligomeric alkyl cyclohexylcarbodiimide (OACC) reagent via ROM polymerization from commercially available starting materials is described. This reagent is exploited as a coupling reagent for esterification, amidation, and dehydration of carboxylic acids (aliphatic and aromatic) with an assortment of alcohols (aliphatic primary, secondary, and benzylic), thiols, phenols, and amines (aliphatic primary, secondary, benzylic, and aromatic/anilines), respectively. Following the coupling event, precipitation with an appropriate solvent (Et(2)O, MeOH, or EtOAc), followed by filtration through a SPE provides the products in good to excellent yield and purity. PMID- 15549806 TI - Ion pair first and second acidities of some beta-diketones and aggregation of their lithium and cesium enediolates in THF. AB - Ion pair pK values were measured for three beta-diketones in THF, 1-3, with lithium and cesium counterions. The results showed variations with concentration indicative of aggregation of the metal enolates to dimers. Similarly, ion pair pK values could be determined for some of these metal enolates going to the corresponding dimetal dienediolates which were also found to form dimers. These equilibria are more complicated to analyze because aggregation affects both sides of the proton transfer equilibria. The results show that all of the species measured exist mostly as dimers at concentrations >0.01 M typical of most organic synthesis reactions and physical measurements. NMR measurements show that the enols of 1 and 2, which can undergo intramolecular hydrogen bonding, predominate in both THF and DMSO solutions, whereas 3, whose enols cannot be so stabilized, is mostly keto in THF but approximately equimolar enol and keto in DMSO. Dimerization of the monolithium salts is rapid on the NMR time scale but that of the dilithium salts is slow. PMID- 15549808 TI - Application of bis(iminophosphorane) in heterocyclic synthesis: new entries to symmetrically or unsymmetrically substituted thieno[2,3-d:5,4-d']dipyrimidine 4,5(3H,6H)-diones. AB - The bis(carbodiimides) 4, obtained from bis-aza-Wittig reactions of bis(iminophosphorane) 3 with 2 equiv of aromatic isocyanates, were reacted with secondary amine to give symmetrically substituted 2,7-diaminothieno[2,3-d:5,4 d']dipyrimidine-4,5(3H,6H)-dione 6 in the presence of a catalytic amount of EtO( )Na(+). Reactions of 4 with phenols or ROH in the presence of a catalytic amount of potassium carbonate or RO(-)Na(+) gave symmetrically substituted 2,7 diaryl(alkyl)oxythieno[2,3-d:5,4-d']dipyrimidine-4,5(3H,6H)-diones 6 in satisfactory yields. However, iminophosphoranes 9 were obtained via reaction of bis(iminophosphorane) 3 with 1 equiv of aromatic isocyanate and subsequent reaction with an amine in the presence of a catalytic amount of EtO(-)Na(+). Further reaction of iminophosphoranes 9 with aromatic isocyanates and various nucleophile generated unsymmetrically substituted thieno[2,3-d:5,4 d']dipyrimidine-4,5(3H,6H)-diones 12 in good yields. PMID- 15549807 TI - 9-Acylation of 1-acyldipyrromethanes containing a dialkylboron mask for the alpha acylpyrrole motif. AB - 1,9-Diacyldipyrromethanes are important precursors to porphyrins, yet synthetic access remains limited owing to (1) poor conversion in the 9-acylation of 1 acyldipyrromethanes and (2) handling difficulties because acyldipyrromethanes typically streak upon chromatography and give amorphous powders upon attempted crystallization. A reliable means for converting a dipyrromethane to a 1 acyldipyrromethane-dialkylboron complex was recently developed, where the dialkylboron (BR(2)) unit renders the complex hydrophobic and thereby facilitates isolation. Herein a refined preparation of 1,9-diacyldipyrromethanes is presented that employs the 1-acyldipyrromethane-BR(2) complex as a substrate for 9 acylation. The dialkylboron unit provides protection for the alpha-acylpyrrole unit. 9-Acylation requires formation of the pyrrolyl-MgBr reagent and the presence of 1 equiv of a nonnucleophilic base to quench the proton liberated upon alpha-acylation. Reaction of the 1-acyldipyrromethane-BR(2) complex (1 equiv) with mesitylmagnesium bromide (2 equiv) followed by the addition of an acylating agent (S-2-pyridyl thioate or acid chloride, 1.1 equiv) gives the corresponding 1,9-diacyldipyrromethane-BR(2) complex. The acylation method afforded 1,9 diacyldipyrromethane-BR(2) complexes with limited or no chromatography in yields of 64-92%. The 1,9-diacyldipyrromethane-BR(2) complexes are stable to routine handling, are readily soluble in common organic solvents, crystallize readily, and can now be prepared in multigram quantities through use of stoichiometric quantities of reagents. PMID- 15549809 TI - Sequential aza-Baylis-Hillman/ring closing metathesis/aromatization as a novel route for the synthesis of substituted pyrroles. AB - A new route to diverse 2-substituted-3-methoxycarbonyl pyrroles has been developed. Diverse SES protected alpha-methylene beta-aminoesters were obtained by a 3-component aza-Baylis-Hillman reaction. Diversity arose from the aryl aldehydes which can be used in this reaction. N-Alkylation with allyl bromide under mild conditions provided the corresponding dienes. These substituted dienes were cyclized by ring closing metathesis at room temperature or under microwave activation with Grubbs-type II catalyst to yield SES-protected pyrroline intermediates. The final pyrroles were obtained by base-promoted dehydrodesulfinylation/aromatization. The scope of each of these reactions was explored. PMID- 15549810 TI - New synthetic equivalent of nitromalonaldehyde treatable in organic media. AB - beta-Nitroenamines having a formyl group at the beta-position behave as the synthetic equivalent of unstable nitromalonaldehyde, which is a useful synthon for syntheses of versatile nitro compounds. High solubility of the nitroenamines into general organic solvents enables us to conduct reactions in the organic media accompanied by easy experimental manipulations and considerable safety. When nitroenamines are treated with 1,2-bifunctional nucleophiles such as hydrazines, hydroxylamine and glycine ester, nitrated pyrazoles, isoxazole and pyrrole-2-carboxylate were readily prepared. This methodology was also applicable to guanidines and 1,2-diamines, leading to pyrimidines and 1,4-diazepines, respectively. PMID- 15549811 TI - Highly stereoselective synthesis of functionalized beta,beta-di- and trisubstituted vinylic sulfoxides by Cu-catalyzed conjugate addition of organozinc reagents. AB - beta,beta-Disubstituted chiral vinylic sulfoxides bearing functionalities have been synthesized via Cu-catalyzed conjugate addition of organozinc reagents to chiral 1-alkynyl sulfoxides. Due to the availability of functionalized organozinc reagents and high syn-selectivity of the reaction, both geometric beta,beta disubstituted vinylic sulfoxides were selectively synthesized. Furthermore, 1 alkynyl sulfoxides were derivatized into trisubstituted vinylic sulfoxides by trapping the resulting alpha-sulfinyl vinylic carbanion with electrophiles. Highly diastereoselective THF and THP ring formations were accomplished by means of this methodology followed by an intramolecular Michael addition. PMID- 15549812 TI - Solid-phase synthesis of arginine-containing peptides and fluorogenic substrates using a side-chain anchoring approach. AB - Attachment of an amino acid to a solid support by its side chain is sometimes necessary to take advantage of an alpha-carboxylic group available for diverse modifications, including the incorporation of a fluorophore for the preparation of fluorogenic substrates. In contrast to most other amino acids, anchoring the guanidinium group of an arginine to a resin requires the use of a supplementary linker. To avoid the usually multistep synthesis of such a linker as well as its difficult attachment to the guanidine group, we developed a simple method where the guanidine group is built on a Rink amide resin. Our strategy followed the steps of guanidine formation: (i) addition of an isothiocyanate derivative of ornithine to the amino group of a solid support, yielding Nomega-linked thiocitrulline; (ii) S-methylation of thiourea; (iii) guanidinylation using ammonium acetate. Cleavage of the resin generated the arginine-containing compound, the amine group of the resin becoming part of the guanidine. We have demonstrated the usefulness of this method by the synthesis of a series of fluorogenic substrates for trypsin-like serine proteases, which were obtained in high yield and purity. Then, our strategy also allowed generation from the same precursor differentially substituted arginine derivatives, including Nomega methyl- and Nomega-ethylarginines. The ability to prepare such analogues together with the intermediates thiocitrulline and S-methylisothiocitrulline from a unique precursor while the alpha-amine and carboxylic groups remain available for modification also makes this method a powerful tool for combinatorial solid-phase synthesis of NO synthase inhibitors. PMID- 15549813 TI - Diastereoselective photochromism of bisbenzothienylethenes with an oxycarbonyl related functional group on the side chain. AB - Nine 1-[2-(1-substituted)ethyl-3-benzothienyl]-2-(2-methyl-3 benzothienyl)hexafluorocyclopentenes (4-substituted benzoyloxy, N arylcarbamoyloxy, and aryloxycarbonyloxy) were synthesized. They showed diastereoselective photochromic ring closure upon UV irradiation. Among them, 4 nitrophenoxycarbonyloxy-substituted compound recorded a remarkable 94% diastereomer excess with 91% conversion to the closed form. PMID- 15549814 TI - Nickel-catalyzed addition of alkenylzirconium reagents to bicyclic olefins: a highly regio- and stereoselective ring-opening reaction. AB - A highly regio- and stereoselective ring-opening addition of alkenylzirconium reagents to bicyclic olefins catalyzed by nickel complexes was described. Treatment of 7-oxa- and 7-azabenzonorbornadienes (1a-e) with various terminal alkenylzirconium reagents 2a-f (Cp(2)ZrClCH=CHR; R = t-Bu, n-Pr, n-Oct, 1 cyclohexenyl, SiMe(3), and Ph) in the presence of Ni(PPh(3))(2)Cl(2) and Zn powder (or a combination of ZnCl(2) and NEt(3)) in dry THF at 50 degrees C afforded the corresponding cis-2-alkenyl-1,2-dihydronaphthalene derivatives 3a-l in moderate to excellent yields. Under similar reaction conditions, internal alkenylzirconium reagents 2g,h (Cp(2)ZrClCR=CHR: R = Et and n-Pr) also undergo ring-opening addition to oxanorbornadienes 1a and 1d to give cis-2-alkenyl-1,2 dihydronaphthalene derivatives 4a-c in good yields. Possible pathways involving the transfer of alkenyl group in the alkenylzirconium reagent to the Ni(II) center followed by migration of the alkenyl group from the Ni(II) center to the carbon-carbon double bond of 7-oxanorbornadiene or the reaction of 7 oxanorbornadiene with Ni(0) to form a Ni(II)-pi-allyl prior to the transfer of the alkenyl group as key steps for the catalytic reaction were proposed and discussed. PMID- 15549815 TI - Efficient Baylis-Hillman reactions of cyclic enones in methanol as catalyzed by methoxide anion. AB - The Baylis-Hillman reactions of cyclic enones with a variety of aldehydes were investigated. 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU) was found to be a viable catalyst in promoting the reactions of sterically retarded substrates in methanol. The reactions showed clear solvent dependence and only occurred in hydroxylic solvents, especially in methanol. Further consideration on the steric character of DBU and its high basicity jointly with other experimental observations suggests that the methoxide anion should be the "true" Baylis Hillman catalyst. This has been confirmed by the effectiveness of similar reactions directly employing methoxide as the catalyst. The reaction pathways of this type of catalysis are proposed to depend on the choice of substrates. Supporting experimental observations were demonstrated and discussed in relation to mechanistic considerations. This study also reveals that both DBU and sodium methoxide can be successfully applied as effective catalysts in methanol to promote the Baylis-Hillman reactions for a range of cyclic enones including cyclopent-2-enones, cyclohex-2-enones, gamma-pyrone, and 1-benzopyran-4(4H)-ones. PMID- 15549816 TI - Superoxide dismutase versus ferricytochrome C: determining rate constants for the spin trapping of superoxide by cyclic nitrones. AB - Given that spin trapping/electron paramagnetic resonance (EPR) spectroscopy has become the primary technique to identify important biologically generated free radicals, such as superoxide (O(2)(*-)), in vitro and in vivo models, evaluation of the efficiency of specific spin traps to identify this free radical is paramount. Recently, a family of ester-containing nitrones has been prepared, which appears to have distinct advantages for spin trapping O(2)(*-) compared to the well-studied spin traps 5,5-dimethyl-1-pyrroline N-oxide 1 and 5 (diethoxyphosphoryl)-5-methyl-1-pyrroline N-oxide 2. An important determinant in the selection of a spin trap is the rate constant (k(app)) for its reaction with O(2)(*-), and several different methods have been employed in estimating this k(app). In this paper, the two most frequently used scavengers of O(2)(*-), ferricytochrome c and Cu/Zn-SOD, were evaluated as competitive inhibitors for spin trapping this free radical. Data presented herein demonstrate that SOD is the preferred compound when determining the k(app) for the reaction of O(2)(*-) with spin traps. Using this model, the k(app) for the reaction of nitrone 1, 5 tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide 3, and 5-methoxycarbonyl-5 methyl-1-pyrroline N-oxide 4 with O(2)(*)(-) was estimated to be 24.6 +/- 3.1, 73.0 +/- 12, and 89.4 +/- 1.0 M(-1) s(-1) at pH 7.0, respectively. Several other comparative studies between known spin traps were also undertaken. PMID- 15549817 TI - Design of a nonreductive method for chemoselective cleavage of hydrazines in the presence of unsaturations: application to a stereoconvergent three-component synthesis of (-)-methyl palustramate. AB - A chemoselective hydrazine (N-N) cleavage methodology that preserves the integrity of alkenes was developed based on a mild acid-promoted fragmentation of tetrasubstituted 1-(trimethylsilylmethyl)-1-benzylhydrazines. This strategy was applied to a concise asymmetric synthesis of (-)-methyl palustramate (4), which featured a convergent stereo- and regioselective sequential three-component aza[4+2]cycloaddition/allylboration/retro-sulfinyl-ene rearrangement between diene 1f, dienophile 2b, and propionaldehyde to afford cis-2-carboxy-6 hydroxyalkylpiperidine 25. The acid-promoted hydrazinolysis of 25 cleanly afforded key intermediate 31, and the latter led to target 4 in four steps after a series of functional group transformations. PMID- 15549818 TI - Synthesis of highly functionalized chiral nitriles by radical fragmentation of beta-hydroxy azides. Convenient transformation of aldononitriles into 1,4- and 1,5-iminoalditols. AB - The synthesis of highly functionalized nitriles by an alkoxyl radical fragmentation of cyclic beta-hydroxy azides is described. The alkoxyl radicals were generated by reaction of the alcohols with (diacetoxyiodo)benzene and iodine under mild conditions compatible with the presence of sensitive substituents and the protective groups most frequently used in carbohydrate chemistry. To explore the scope and limitations of this methodology, experiments were carried out using a variety of beta-hydroxy azides of the carbohydrate (1-6, 33, and 41), monoterpenoid (21 and 22), and steroid (23-25) families of natural products. Of special interest are the aldopentonitriles (15-18, 34, and 42) and aldotetrononitriles (19 and 20) synthesized from the corresponding 2-azido-2 deoxycarbohydrates. To demonstrate the versatility of these aldononitriles as chiral synthons, 1,4-imino-1-deoxysugar (37) and 1,5-imino-1-deoxysugar (43) analogues of the polyhydroxypyrrolidine and -piperidine types were prepared. PMID- 15549819 TI - Palladium-catalyzed [2 + 2 + 2] cocyclotrimerization of benzynes with bicyclic alkenes: an efficient route to anellated 9,10-dihydrophenanthrene derivatives and polyaromatic compounds. AB - An efficient method for the cocyclotrimerization of bicyclic alkenes and benzynes catalyzed by palladium phosphine complexes to give the corresponding norbornane anellated 9,10-dihydrophenanthrene derivatives is described. Bicyclic alkenes 1a i undergo [2 + 2 + 2] cocyclotrimerization with benzynes generated from precursors 2a-d [2-(trimethylsilyl)phenyl triflate (2a), 4,5-dimethyl-2 (trimethylsilyl)phenyl triflate (2b), 6-(trimethylsilyl)-2,3-dihydro-1H-5-indenyl triflate (2c), 4-methyl-2-(trimethylsilyl)phenyl triflate (2d)] in the presence of PdCl(2)(PPh(3))(2) in acetonitrile at ambient temperature to yield anellated 9,10-dihydrophenanthrene products 3a-r in moderate to excellent yields. The [2 + 2 + 2] cocyclotrimerization products from oxa- and azabicyclic alkenes can be applied for the synthesis of polyaromatics, substituted benzo[b]triphenylenes (8a f), via a simple Lewis acid mediated deoxyaromatization in good yields. In addition the [2 + 2 + 2] products undergo retro Diels-Alder reaction readily, providing a new method for the synthesis of substituted phenanthrenes and for generating isobenzofurans. A plausible mechanism is proposed to account for the catalytic [2 + 2 + 2] cycloaddition reaction. PMID- 15549820 TI - Reaction pathways and free energy barriers for alkaline hydrolysis of insecticide 2-trimethylammonioethyl methylphosphonofluoridate and related organophosphorus compounds: electrostatic and steric effects. AB - Reaction pathways and free energy barriers for alkaline hydrolysis of the highly neurotoxic insecticide 2-trimethylammonioethyl methylphosphonofluoridate and related organophosphorus compounds were studied by performing first-principles electronic structure calculations on representative methylphosphonofluoridates, (RO)CH3P(O)F, in which R = CH2CH2N+(CH3)3, CH3, CH2CH2C(CH3)3, CH2CH2CH(CH3)2, CH(CH3)CH2N+(CH3)3, and CH(CH3)CH2N(CH3)2. The dominant reaction pathway was found to be associated with a transition state in which the attacking nucleophile OH- and the leaving group F- are positioned on opposite sides of the plane formed by the three remaining atoms attached to the phosphorus in order to minimize the electrostatic repulsion between these two groups. The free energy barriers calculated for the rate-determining step of the dominant pathway are 12.5 kcal/mol when R = CH2CH2N+(CH3)3, 15.5 kcal/mol when R = CH3, 17.9 kcal/mol when R = CH2CH2C(CH3)3, 16.5 kcal/mol when R = CH2CH2CH(CH3)2, 13.4 kcal/mol when R = CH(CH3)CH2N+(CH3)3, and 18.7 kcal/mol when R = CH(CH(3))CH(2)N(CH(3))(2). The calculated free energy barriers are in good agreement with available experimentally derived activation free energies, i.e. 14.7 kcal/mol when R = CH(3), 13.4 kcal/mol when R = CH2CH2N+(CH3)3, and 13.9 kcal/mol when R = CH(CH3)CH2N+(CH3)3. A detailed analysis of the calculated energetic results and available experimental data suggests that the net charge of the molecule (M) being hydrolyzed is a prominent factor affecting the free energy barrier (DeltaG) for the alkaline hydrolysis of phosphodiesters, phosphonofluoridates, and related organophosphorus compounds. The electrostatic interactions between the attacking nucleophile OH- and the molecule M being hydrolyzed favor such an order of the free energy barrier: DeltaG(M(+)+OH-) < DeltaG(M0+OH-) < DeltaG(M(-)+OH-), where M+, M0, and M- represent the cationic, neutral, and anionic molecules, respectively. The change of the substituent R in (RO)CH(3)P(O)F from CH3 to CH2CH2N+(CH3)3 is associated with both the electrostatic and steric effects on the free energy barrier, but the electrostatic effect dominates the substituent shift of the free energy barrier. This helps to better understand why the alkaline hydrolysis of (RO)CH3P(O)F with R = CH2CH2N+(CH3)3 and CH(CH3)CH2N+(CH3)3 is significantly faster than that with R = CH3. The effect of electrostatic interaction also helps to understand why the rate constants for the alkaline hydrolysis of phosphodiesters, such as intramolecular second messenger adenosine 3',5'-phosphate (cAMP), are generally smaller than those for the alkaline hydrolysis of the phosphonofluoridates and related phosphotriesters. PMID- 15549821 TI - Highly diastereoselective tandem photoenolization-hetero-Diels-Alder cycloaddition reactions of o-tolualdehydes in the solid state. AB - The (E)-photoenols generated in situ by photolysis of o-tolualdehydes 1-5 in the solid state react with the precursor aldehydes as dienophiles in a hetero-Diels Alder cycloaddition fashion to afford trans-3-arylisochromanols in excellent yields and in a high diastereoselectivity. An examination of the reactivity of three different classes of rationally designed aldehydes shows that the tandem enolization-Diels-Alder cycloaddition occurs in the solid state efficiently for aldehydes whose (E)-photoenols (i) are more stable than their corresponding benzocyclobutenols and (ii) are not sterically congested. However, rapid cyclization to benzocyclobutenols is found to be the sole pathway for sterically encumbered (E)-enols derived from aldehydes 6-8. Given that the execution of heteromolecular reactions in the solid state is a challenge, the results obtained with simple crystalline o-tolualdehydes are remarkable and the strategy involving in situ generation of a dienol in the crystal lattice of a dienophile to achieve hetero-Diels-Alder reaction in a highly diastereoselective manner is heretofore unprecedented. In the context of enormous interest in tandem/domino reactions in contemporary synthetic organic chemistry, the results observed with o tolualdehydes exemplify successful execution of tandem reactions in the solid state. PMID- 15549822 TI - Ruthenium-catalyzed [2 + 2] cycloadditions between 7-substituted norbornadienes and alkynes: an experimental and theoretical study. AB - The ruthenium-catalyzed [2 + 2] cycloadditions of 7-substituted norbornadienes with an alkyne have been investigated. The cycloadditions were found to be highly regio- and stereoselective, giving only the anti-exo cycloadducts as the single regio- and stereoisomers in good yields. The results on the relative rate of different 7-substituted norbornadienes in the Ru-catalyzed [2 + 2] cycloadditions with an alkyne indicated that the reactivity of the alkene component decreases dramatically as the alkene becomes more electron deficient. Ab initio computational studies on the ruthenium-catalyzed [2 + 2] cycloadditions provided important information about the geometries and the arrangements of the four different groups on the Ru in the initial Ru-alkene-alkyne pi-complex, 14, and in the metallacyclopentene 15. Based on our computational studies, we also found that the first carbon-carbon bond formed in the [2 + 2] cycloaddition is between the C(5) of the alkene and the C(b) (the acetylenic carbon attached to the ester group) of the alkyne 8. Our computational studies on the potential energy profiles of the cycloadditions showed that the activation energy relative to the reactants for the oxidative addition step is in the range of 9.3-9.8 kcal/mol. The activation energy relative to the metallacyclopentene for the reductive elimination step is much higher than for the oxidative addition step (in the range of 25.9-27.6 kcal/mol). PMID- 15549823 TI - A computational study of the Diels-Alder reaction of ethyl-S-lactyl acrylate and cyclopentadiene. Origins of stereoselectivity. AB - The eight diastereoisomeric transition structures of the Diels-Alder addition of ethyl-S-lactyl acrylate and cyclopentadiene have been investigated in the gas phase and in solution by HF, MP2, and density functional theory (B3LYP and B3PW91) methods with the 6-31G(d,p) basis set. At all levels of theory used, the s-cis transition structures are more stable than the s-trans ones. The contribution of the s-trans transition structures increases in solution and, although still small, has to be taken in consideration for correct prediction of stereoselectivity. Diastereofacial selectivity is interpreted in terms of electrostatic (weak hydrogen bonding) C=O...H(C) interactions between the carbonyl group(s) of the dienophile and cyclopentadiene in the energetically favored transition structures. Endo/exo reaction selectivity is attributed to positive orbital interactions between the diene and the acrylate carbonyl oxygen in the endo s-cis transition structures. Ab initio methods reproduce well the experimentally observed trends in both endo/exo and diastereofacial selectivity. Density functional calculations in the gas phase correctly reproduce the observed trends in diastereofacial selectivity but single-point MP2 calculations are necessary to reproduce the experimental trend in endo/exo selectivity. PMID- 15549824 TI - Transition metal salt-catalyzed direct three-component mannich reactions of aldehydes, ketones, and carbamates: efficient synthesis of N-protected beta-aryl beta-amino ketone compounds. AB - The transition metal salt-catalyzed direct three-component Mannich reactions of aryl aldehydes, aryl ketones, and carbamates are described. The RuCl(3).xH(2)O-, AuCl(3)-PPh(3)-, and AuCl(3)-catalyzed direct Mannich reactions led to the synthesis of N-protected beta-aryl-beta-amino ketones, and the results create new possibilities for exploiting the transition metal salt-catalyzed direct Mannich reaction and facile synthesis of beta-amino ketone libraries. PMID- 15549825 TI - New C-C coupling reaction of cyclic nitronates with carbon nucleophiles. Umpolung of the conventional reactivity of nitronates. AB - Cyclic six-membered nitronates 1 are involved in diastereoselective C-C coupling reactions with various nucleophiles in the presence of either catalytic or stoichiometric amounts of TBDMSOTf to give the previously unknown N siloxytetrahydrooxazines. The intermediacy of N,N-bis(oxy)iminium cations was proven by NMR data. PMID- 15549826 TI - Synthesis of symmetric bis(imidazole-4,5-dicarboxamides) substituted with amino acids. AB - A series of symmetric bis(imidazole-4,5-dicarboxamides) (bis-I45DCs) were prepared with amino acid esters and a variety of linker groups. The critical pyrazine intermediates, substituted with amino acid esters, were synthesized by stoichiometric control of the amino acid ester, even though primary alkanamines, in comparison, generally offer less selectivity for this reaction. Diamines are added to subsequently react with and open the remaining acyl imidazole bonds in the pyrazine intermediates and thereby yield the bis-I45DCs. PMID- 15549827 TI - Synthesis of 4'-methyl and 4'-cyano carbocyclic 2',3'-didehydro nucleoside analogues via 1,4-addition to substituted cyclopentenones. AB - Carbocyclic 4'-methyl and 4'-cyano nucleoside analogues were synthesized using the Michael reaction to introduce the 4'-substituent and Pd-catalyzed allylic substitution to introduce the nucleoside base. Use of both the desired beta- and undesired alpha-1'-carbonate diastereomers in the Pd-catalyzed substitution was demonstrated in principle by epimerization of the alpha-diastereomer and kinetic diastereodifferentiation of a 1:1 alpha/beta mixture of 1'-carbonates. PMID- 15549828 TI - An unexpected three-component condensation leading to amino- (3-oxo-2,3-dihydro 1H-isoindol-1-ylidene)- acetonitriles. AB - The reaction of 2-carboxybenzaldehyde with primary amines in the presence of cyanide leads to the formation of 2-substituted amino(3-oxo-2,3-dihydro-1H isoindol-1-ylidene)acetonitriles. These compounds originate from the condensation of 2-carboxybenzaldehyde with the amine and two molecules of hydrogen cyanide and represent a novel class of isoindolinones. PMID- 15549830 TI - Guanidines as a nitrogen source for direct conversion of epoxides to aziridines. AB - Direct conversion of epoxides to aziridines was achieved with guanidines as a nitrogen source. Stereochemical inversion at the chiral centers of epoxides was observed without loss of optical purity. PMID- 15549829 TI - Efficient asymmetric oxidation of sulfides and kinetic resolution of sulfoxides catalyzed by a vanadium-salan system. AB - The asymmetric oxidation of sulfides to chiral sulfoxides with hydrogen peroxide in good yield and high enantioselectivity has been catalyzed very effectively by chiral vanadium-salan [N,N'-alkyl bis(salicylamine)] complex. The salan ligand shows results superior in terms of reactivity and enantioselectivity to those of salen [N,N'-alkylene bis(salicylideneimine)] analogue, and provides the sulfoxide with opposite configuration. The high enantioselectivity of this reaction is the direct result of the asymmetric oxidation. The efficient kinetic resolution of racemic sulfoxides catalyzed by the vanadium-salan system is also described. PMID- 15549831 TI - A convenient synthesis of alpha-methylene-gamma-butyrolactones from allenyl carbonyl units mediated by Mo(CO)6 through intramolecular cyclocarbonylation. AB - A novel procedure for the synthesis of cis-fused bicyclic alpha-methylene-gamma butyrolactones from allenyl carbonyl functionalities via the molybdenum-mediated cyclocarbonylation is described. The use of Mo(CO)(6) with DMSO to promote reaction results in an efficient and convenient protocol through the three component assembly in high yields. PMID- 15549832 TI - Concerning the efficient conversion of epoxy alcohols into epoxy ketones using dioxiranes. AB - Representative epoxy alcohols are cleanly converted into the corresponding epoxy ketones in high yield by selective oxidation using dimethyldioxirane (1a) and its trifluoro analogue (1b) under mild conditions. The oxidation is found to take place leaving the configuration at the epoxy functionality unaffected. The direct oxyfunctionalization of simple cyclic epoxides with the powerful dioxirane 1b provides another attractive method to access epoxy ketones regioselectively. PMID- 15549833 TI - Indirect trapping of the retroconjugate addition reaction intermediate involved in the epimerization of lobeline: application to the synthesis of (-)-sedamine. AB - Alkyl chloroformates induced indirect trapping of the retroconjugate addition reaction intermediate involved in the epimerization of lobeline is described. This strategy was applied to the conversion of (-)-lobeline to (-)-sedamine in high overall yield. PMID- 15549834 TI - Palladium-catalyzed N-vinylation of sulfoximines. AB - New previously unavailable N-vinyl sulfoximines have been synthesized by intermolecular palladium-catalyzed coupling between sulfoximines and vinyl bromides in excellent yield. Hydrogenation of the vinyl moiety opens a novel way to alpha-branched N-alkyl sulfoximines. PMID- 15549835 TI - Large-scale synthesis and resolution of TRISPHAT [tris(tetrachlorobenzenediolato) phosphate(V)] anion. AB - Both enantiomers of TRISPHAT anion can be obtained on a multigram scale through a novel resolution procedure. The Lambda enantiomer is isolated as the tri-n butylammonium salt, [(n)()Bu(3)NH][Lambda-1], which is soluble in pure CHCl(3) and CH(2)Cl(2). The Delta enantiomer is prepared as the cinchonidinium derivative, which is only soluble in polar solvent mixtures (>7.5% DMSO in CHCl(3)). PMID- 15549836 TI - Synthesis of alpha-hydrazino ketones via regio- and stereoselective electrophilic amination of manganese enolates and enamines. AB - A straightforward procedure for the regio- and stereoselective synthesis of alpha hydrazino ketones is described. Manganese enolates and manganese enamines derived from ketones and from the corresponding N-sulfinylimines react with azodicarboxylate esters (DTBAD and DEAD) in a regioselective fashion to afford in good to excellent yields the kinetic alpha-hydrazino ketones as sole or highly prevalent products. When enantiopure N-sulfinyl manganese enamines were used the stereoselectivity of these reactions ranged from 40% to 68% ee. PMID- 15549837 TI - The preparation of (1alpha,3beta)-3-hydroxycholestane-4,6-diene- 1,25-diol diacetate from a 5,7-diene precursor: a new method for the synthesis of heteroannular dienes. AB - Starting from the 7alpha-bromide 5a, a regioselective synthesis of (1alpha,3beta) 3-hydroxycholestane-4,6-diene-1,25-diol diacetate (2) is described. The preparative removal of contaminating 5,7-diene 9 was accomplished by the formation of the corresponding Diels-Alder adduct 11. Acetylation of the diacetate 2 followed by acid-catalyzed elimination and rearrangement yielded styrene 13. PMID- 15549838 TI - Synthesis of enantiopure tert-butanesulfinamide from tert butanesulfinyloxazolidinone. AB - A three-step procedure for the preparation of enantiopure tert-butanesulfinamide 6 in 51% overall yield is described starting from (1R,2S)-N-Cbz-1,2 diphenylaminoethanol. The key step is the reaction of tert-butylmagnesium chloride with N-Cbz-4,5-diphenyl-1,2,3-oxathiazolidine-2-oxide 2 to afford the optical pure tert-butylsulfinyl-4,5-diphenyl-1,3-oxazolidinone 5 via an 1,5 alkoxy anion rearrangement, which is then subject to ammonia hydrolysis with LiNH(2) in liquid ammonia to give (R)-tert-butanesulfinamide 6. PMID- 15549839 TI - Lewis Acid catalyzed dipolar cycloadditions of an activated imidate. AB - An evaluation of simple Lewis acids revealed that N-malonylimidates undergo catalyzed [3+2] cycloaddition reactions with aldehydes, imines, and activated olefins to form oxazolines, imidazolines, and pyrrolines, respectively. Reactions proceed optimally at ambient temperature with the addition of 5 mol % of MgCl(2) in CH(3)CN. Experiments aimed at elucidation of the reactive intermediate undergoing cycloaddition suggest that the Lewis acid promotes a 1,2-prototropic shift to give a metal-coordinated azomethine ylide, rather than ionization and proton transfer to give a nitrile ylide. PMID- 15549840 TI - Skeletal reorganization of enynes into 1-vinylcycloalkenes in ionic liquids. AB - The skeletal reorganization of enynes catalyzed by transition metal chlorides, such as PtCl(2), [RuCl(2)(CO)(3)](2), [RhCl(CO)(2)](2), and AuCl(3), in ionic liquids proceeds under milder conditions (at lower reaction temperatures and for shorter reaction times) than those needed for ordinary solvents. The products produced by the skeletal reorganization of enynes were easily removed from the catalyst by a simple extraction with Et(2)O or distillation. The PtCl(2) can be reused up to five times. PMID- 15549841 TI - New method to prepare peptide-oligonucleotide conjugates through glyoxylic oxime formation. AB - A new method to prepare peptide-oligonucleotide conjugates through chemoselective glyoxylic oxime linkage is reported. A novel phosphoramidite reagent, readily accessible from serine, was prepared and used in automated DNA synthesis to prepare oligonucleotides carrying a glyoxylic aldehyde functionality at the 5' terminus. This was efficiently coupled to a peptide functionalized with an aminooxy group. The method could be of general use to prepare a broad range of oligonucleotide conjugates. PMID- 15549842 TI - Synthesis of 1,1,4,4-tetrabromo-2-butenes and related compounds via desilylation bromination of silylated 1,3-butadiene derivatives. AB - The combination of zirconocene-mediated coupling of silylated alkynes with a protonation-desilylation or bromination-desilylation process afforded otherwise unavailable butadiene derivatives. When (E,E)-2,3-dialkyl-1,4-bis(trimethylsilyl) 1,3-butadienes were treated with 3 equiv of Br(2) in CH(2)Cl(2), (E)-2,3-dialkyl 1,1,4,4-tetrabromo-2-butenes were obtained in excellent yields with perfect stereoselectivity. PMID- 15549843 TI - New tin templates for the synthesis of macrocyclic polythiaether-polythiaester ligands. AB - The preparation of new tin templates, stannathianes 1-3 is described. New templates have been successfully applied to the synthesis of macrocyclic polythialactones 4-9 by cyclization of corresponding stannathianes 1-3 with pimeloyl dichloride. PMID- 15549844 TI - Magnesium iodide promoted reactions of nitrones with cyclopropanes: a synthesis of tetrahydro-1,2-oxazines. AB - Anhydrous magnesium iodide (MgI(2)) is shown to be an effective promoter of the "homo 3+2" dipolar cycloaddition of nitrones with 1,1-cyclopropane diesters. In almost all cases the products tetrahydro-1,2-oxazines are formed in excellent yields. The reactions are highly diastereoselective for a cis relationship between the substitutents at the 3- and 6-positions on the tetrahydrooxazine ring. As an alternative to using a preformed nitrone, the reaction may be performed in a 3-component sense by combining an aldehyde, an hydroxylamine, and the cyclopropane in the presence of catalytic MgI(2). PMID- 15549845 TI - Elaboration of D-(-)-ribose into a tricyclic, natural product-like scaffold. AB - The construction of natural product-like, tricyclic compounds is reported. Starting from a D-(-)-ribose-derived dihydrofurane, the tricyclic scaffold is prepared via an intramolecular hetero-Diels-Alder reaction. The reaction proceeds with very high diastereoselectivity through an endo transition state, as established on the basis of X-ray structural analysis of the products. Further modification and derivatization of the obtained products is described. PMID- 15549846 TI - Structure-dependent oxidative bromination of unsaturated C-C bonds mediated by selectfluor. AB - A number of olefins were subjected to oxidative bromination using Selectfluor/KBr. For different types of substrates, addition, monobromine substituted, or Hunsdiecker-Borodin reaction products can be readily afforded. PMID- 15549849 TI - Strong light fields coax intramolecular reactions on femtosecond time scales. AB - Energetic H(2) (+) ions are formed as a result of intramolecular rearrangement during fragmentation of linear alcohols (methanol, ethanol, propanol, hexanol, and dodecanol) induced by intense, pulsed optical fields. The laser intensity regime that is accessed in these experiments (peak intensity of 8 x 10(15) W cm( 2)) ensures multiple ionization of the irradiated alcohol molecules such that Coulomb explosions would be expected to dominate the overall fragmentation dynamics. Polarization dependent measurements show, counterintuitively, that rearrangement is induced by the strong optical field within a single, 100 fs long laser pulse, and that it occurs before Coulomb explosion of the field-ionized multiply charged alcohols. PMID- 15549850 TI - Molecular potential energy surfaces by interpolation: strategies for faster convergence. AB - A method for interpolating molecular potential energy surfaces introduced [Ischtwan and Collins, J. Chem. Phys. 100, 8080 (1994)] and developed as an iterative scheme has been improved by different criteria for the selection of the data points. Refinements in the selection procedure are based on the variance of the interpolation and the direct exploration of the interpolation error, and produce more accurate surfaces than the previously established scheme for the same number of data points. PMID- 15549851 TI - Comparison of methods for finding saddle points without knowledge of the final states. AB - Within the harmonic approximation to transition state theory, the biggest challenge involved in finding the mechanism or rate of transitions is the location of the relevant saddle points on the multidimensional potential energy surface. The saddle point search is particularly challenging when the final state of the transition is not specified. In this article we report on a comparison of several methods for locating saddle points under these conditions and compare, in particular, the well-established rational function optimization (RFO) methods using either exact or approximate Hessians with the more recently proposed minimum mode following methods where only the minimum eigenvalue mode is found, either by the dimer or the Lanczos method. A test problem involving transitions in a seven-atom Pt island on a Pt(111) surface using a simple Morse pairwise potential function is used and the number of degrees of freedom varied by varying the number of movable atoms. In the full system, 175 atoms can move so 525 degrees of freedom need to be optimized to find the saddle points. For testing purposes, we have also restricted the number of movable atoms to 7 and 1. Our results indicate that if attempting to make a map of all relevant saddle points for a large system (as would be necessary when simulating the long time scale evolution of a thermal system) the minimum mode following methods are preferred. The minimum mode following methods are also more efficient when searching for the lowest saddle points in a large system, and if the force can be obtained cheaply. However, if only the lowest saddle points are sought and the calculation of the force is expensive but a good approximation for the Hessian at the starting position of the search can be obtained at low cost, then the RFO approaches employing an approximate Hessian represent the preferred choice. For small and medium sized systems where the force is expensive to calculate, the RFO approaches employing an approximate Hessian is also the more efficient, but when the force and Hessian can be obtained cheaply and only the lowest saddle points are sought the RFO approach using an exact Hessian is the better choice. These conclusions have been reached based on a comparison of the total computational effort needed to find the saddle points and the number of saddle points found for each of the methods. The RFO methods do not perform very well with respect to the latter aspect, but starting the searches further away from the initial minimum or using the hybrid RFO version presented here improves this behavior considerably in most cases. PMID- 15549852 TI - Scaled opposite-spin second order Moller-Plesset correlation energy: an economical electronic structure method. AB - A simplified approach to treating the electron correlation energy is suggested in which only the alpha-beta component of the second order Moller-Plesset energy is evaluated, and then scaled by an empirical factor which is suggested to be 1.3. This scaled opposite-spin second order energy (SOS-MP2), where MP2 is Moller Plesset theory, yields results for relative energies and derivative properties that are statistically improved over the conventional MP2 method. Furthermore, the SOS-MP2 energy can be evaluated without the fifth order computational steps associated with MP2 theory, even without exploiting any spatial locality. A fourth order algorithm is given for evaluating the opposite spin MP2 energy using auxiliary basis expansions, and a Laplace approach, and timing comparisons are given. PMID- 15549853 TI - Real-time linear response for time-dependent density-functional theory. AB - We present a linear-response approach for time-dependent density-functional theories using time-adiabatic functionals. The resulting theory can be performed both in the time and in the frequency domain. The derivation considers an impulsive perturbation after which the Kohn-Sham orbitals develop in time autonomously. The equation describing the evolution is not strictly linear in the wave function representation. Only after going into a symplectic real-spinor representation does the linearity make itself explicit. For performing the numerical integration of the resulting equations, yielding the linear response in time, we develop a modified Chebyshev expansion approach. The frequency domain is easily accessible as well by changing the coefficients of the Chebyshev polynomial, yielding the expansion of a formal symplectic Green's operator. PMID- 15549854 TI - Parallel replica dynamics with a heterogeneous distribution of barriers: application to n-hexadecane pyrolysis. AB - Parallel replica dynamics simulation methods appropriate for the simulation of chemical reactions in molecular systems with many conformational degrees of freedom have been developed and applied to study the microsecond-scale pyrolysis of n-hexadecane in the temperature range of 2100-2500 K. The algorithm uses a transition detection scheme that is based on molecular topology, rather than energetic basins. This algorithm allows efficient parallelization of small systems even when using more processors than particles (in contrast to more traditional parallelization algorithms), and even when there are frequent conformational transitions (in contrast to previous implementations of the parallel replica algorithm). The parallel efficiency for pyrolysis initiation reactions was over 90% on 61 processors for this 50-atom system. The parallel replica dynamics technique results in reaction probabilities that are statistically indistinguishable from those obtained from direct molecular dynamics, under conditions where both are feasible, but allows simulations at temperatures as much as 1000 K lower than direct molecular dynamics simulations. The rate of initiation displayed Arrhenius behavior over the entire temperature range, with an activation energy and frequency factor of E(a) = 79.7 kcal/mol and log A/s(-1) = 14.8, respectively, in reasonable agreement with experiment and empirical kinetic models. Several interesting unimolecular reaction mechanisms were observed in simulations of the chain propagation reactions above 2000 K, which are not included in most coarse-grained kinetic models. More studies are needed in order to determine whether these mechanisms are experimentally relevant, or specific to the potential energy surface used. PMID- 15549855 TI - Nonperturbative modeling of two-photon absorption in a three-state system. AB - The physics of the two-photon absorption process is investigated for a three state system. The density-matrix equations for the two-photon interaction are solved in the steady-state limit assuming that the pump laser radiation is monochromatic. Collisional broadening, saturation, and Stark shifting of the two photon resonance are investigated in detail by numerical solution of the steady state density-matrix equations. Analytical expressions for the saturation intensity and the Stark shift are derived for the case where the single-photon transitions between the intermediate state and the initial and final states are far from resonance with the pump laser. For this case, it is found that the direction of the Stark shift is dependent on the relative magnitudes of the dipole-moment matrix elements for the single-photon transitions that couple the intermediate state with the initial and final states. Saturation and Stark shifting are also investigated for the case where the single-photon transitions between the intermediate state and the initial and final states are close to resonance with the pump laser. PMID- 15549856 TI - Spatial distributions of angular momenta in quantum and quasiclassical stereodynamics. AB - We have recently reported a derivation of the relationship between the quantum and classical descriptions of angular momentum polarization [M. P. de Miranda and F. Javier Aoiz, Phys. Rev. Lett. 93, 083201 (2004)]. This paper presents a detailed account of the derivation outlined in that paper, and discusses the implications of the new results. These include (i) a new expression of the role of the uncertainty principle in the broadening of angular momentum distributions, (ii) the attribution of azimuthal fluctuations of angular momentum distributions to spatial quantum beats, (iii) the definition of a new Fourier transform of the density matrix, distinct from those suggested in the past, that provides an alternative view of how the quantum description of angular momentum polarization approaches the classical one in the correspondence principle limit, (iv) a prescription for the determination of a quasiclassical angular momentum distribution function that does not suffer from problems encountered with its purely classical counterpart, and (v) a description of how angular momentum distributions commonly visualized with recourse to the classical vector model can be depicted with exact and well-defined quantum mechanics. PMID- 15549857 TI - Efficiency considerations in the construction of interpolated potential energy surfaces for the calculation of quantum observables by diffusion Monte Carlo. AB - A modified Shepard interpolation scheme is used to construct global potential energy surfaces (PES) in order to calculate quantum observables--vibrationally averaged internal coordinates, fully anharmonic zero-point energies and nuclear radial distribution functions--for a prototypical loosely bound molecular system, the water dimer. The efficiency of PES construction is examined with respect to (a) the method used to sample configurational space, (b) the method used to choose which points to add to the PES data set, and (c) the use of either a one- or two-part weight function. The most efficient method for constructing the PES is found to require a quantum sampling regime, a combination of both h-weight and rms methods for choosing data points and use of the two-part weight function in the interpolation. Using this regime, the quantum diffusion Monte Carlo zero point energy converges to the exact result within addition of 50 data points. The vibrationally averaged O-O distance and O-O radial distribution function, however, converge more slowly and require addition of over 500 data points. The methods presented here are expected to be applicable to both other loosely bound complexes as well as tightly bound molecular species. When combined with high quality ab initio calculations, these methods should be able to accurately characterize the PES of such species. PMID- 15549858 TI - Photoionization and photodissociation dynamics of the B 1sigmau + and C 1Piu states of H2 and D2. AB - The photoionization and photodissociation dynamics of H(2) and D(2) in selected rovibrational levels of the B (1)Sigma(u) (+) and C (1)Pi(u) states have been investigated by velocity map ion imaging. The selected rotational levels of the B (1)Sigma(u) (+) and C (1)Pi(u) states are prepared by three-photon excitation from the ground state. The absorption of fourth photon results in photoionization to produce H(2)(+) X (2)Sigma(g)(+) or photodissociation to produce a ground state H(1s) atom and an excited H atom with n >or= 2. The H(2) (+) ion can be photodissociated by absorption of a fifth photon. The resulting H(+) or D(+) ion images provide information on the vibrational state dependence of the photodissociation angular distribution of the molecular ion. The excited H(n >or= 2) atoms produced by the neutral dissociation process can also be ionized by the absorption of a fifth photon. The resulting ion images provide insight into the excited state branching ratios and angular distributions of the neutral photodissociation process. While the experimental ion images contain information on both the ionic and neutral processes, these can be separated based on constraints imposed on the fragment translational energies. The angular distribution of the rings in the ion images indicates that the neutral dissociation of molecular hydrogen and its isotopes is quite complex, and involves coupling to both doubly excited electronic states and the dissociation continua of singly excited Rydberg states. PMID- 15549859 TI - Transition state dynamics of OHF on several electronic states: photodetachment spectrum of OHF- and conical intersections. AB - Wave packet simulations of the photodetachment spectrum of OHF(-) are performed on several electronic adiabatic states, two triplets and four singlets of neutral OHF. The transition moments to these six states have been approximated using the ab initio electronic wave functions of OHF(-) and OHF calculated at the equilibrium configuration of the parent anion. In a first step, two-dimensional simulations of the spectrum are performed on new two-dimensional potential energy surfaces (PESs) of the neutral in a OHF collinear geometry. The resulting simulated spectrum is in rather good agreement with the experimental one, reproducing all the structures from 0 to 2.5 eV electron kinetic energies. At energies below 0.5 eV, all calculated states, singlets and triplets, contribute to the total spectrum. At higher energies, however, only the triplet states participate. In a second step, to improve the description of the spectrum, three dimensional wave packet simulations of the spectrum are performed, getting an excellent agreement with the experiment. The collinear (3)Sigma(-) and (3)Pi states split in two (3)A(") and one (3)A('). New adiabatic PESs are used in this work for the 2 (3)A(") and 1 (3)A(') states, while the one recently proposed was used for the ground 1 (3)A("). It is found that the minimum energy paths of the (3)Sigma(-) and (3)Pi states cross twice at collinear geometry, so that at the transition state the ground state corresponds to (3)Pi, while (3)Sigma(-) is the lowest state otherwise. Such conical intersections are expected to give rise to important Sigma-Pi vibronic effects, requiring a complete three-dimensional model of coupled diabatic states to improve our understanding of the reaction dynamics in this kind of systems. PMID- 15549860 TI - Relaxation behavior of rovibrationally excited H2 in a rarefied expansion. AB - The evolution of the rotational and vibrational distributions of molecular hydrogen in a hydrogen plasma expansion is measured using laser induced fluorescence in the vacuum-UV range. The evolution of the distributions along the expansion axis shows the relaxation of the molecular hydrogen from the high temperature in the upstream region to the low ambient temperature in the downstream region. During the relaxation, the vibrational distribution, which has been recorded up to v = 6, is almost frozen in the expansion and resembles a Boltzmann distribution at T approximately 2200 K. However, the rotational distributions, which have been recorded up to J = 17 in v = 2 and up to J = 11 in v = 3, cannot be described with a single Boltzmann distribution. In the course of the expansion, the lower rotational levels (J < 5) adapt quickly to the ambient temperature ( approximately 500 K), while the distribution of the higher rotational levels (J > 7) is measured to be frozen in the expansion at a temperature between 2000 and 2500 K. A model based on rotation-translation energy transfer is used to describe the evolution of the rotational distribution of vibrational level v = 2 in the plasma expansion. The behavior of the low rotational levels (J < 5) is described satisfactory. However, the densities of the higher rotational levels decay faster than predicted. PMID- 15549861 TI - Microwave Fourier transform spectrum of the water-carbonyl sulfide complex. AB - The microwave spectrum of the water-carbonyl sulfide complex H(2)O-OCS was observed with a pulsed-beam, Fabry-Perot cavity Fourier-transform microwave spectrometer. In addition to the normal isotopic form, we also measured the spectra of H(2)O-S(13)CO, H(2)O-(34)SCO, H(2) (18)O-SCO, D(2)O-SCO, D(2)O S(13)CO, D(2)O-(34)SCO, HDO-SCO, HDO-S(13)CO, and HDO-(34)SCO. The rotational constants are B = 1522.0115(2) MHz and C = 1514.3302(2) MHz for H(2)O-SCO; B = 1511.9153(5) MHz and C = 1504.3346(5) MHz for H(2)O-S(13)CO; B = 1522.0215(3) MHz and C = 1514.3409(3) MHz for H(2)O-(34)SCO; B = 1435.9571(3) MHz and C = 1429.1296(4) MHz for H(2) (18)O-SCO, B = 1409.6575(5) MHz and C = 1397.9555(5) MHz for D(2)O-SCO; B = 1399.8956(3) MHz and C = 1388.3543(3) MHz for D(2)O S(13)CO; B = 1409.6741(24) MHz and C = 1397.9775(24) MHz for D(2)O-(34)SCO; (B+C)/2 = 1457.9101(2) MHz for HDO-SCO; (B + C)/2 = 1448.0564(4) MHz for HDO S(13)CO; and (B+C)/2 = 1457.9418(15) MHz for HDO-(34)SCO, with uncertainties corresponding to one standard deviation. The observed rotational constants for the sulfur-34 complexes are generally higher than those for the corresponding sulfur-32 isotopomers. The heavier isotopomers have smaller effective moments of inertia due to the smaller vibrational amplitude of the (34)S-C vibration (zero point) as compared to the (32)S-C, making the effective O-(34)S bond slightly shorter. Stark effect measurements for H(2)O-SCO give a dipole moment of 8.875(9)x10(-30) C m [2.6679(28) D]. The most probable structure of H(2)O-SCO is near C(2v) planar with the oxygen of water bonded to the sulfur of carbonyl sulfide. The oxygen-sulfur van der Waals bond length is determined to be 3.138(17) A, which is very close to the ab initio value of 3.144 A. The structures of the isoelectronic complexes H(2)O-SCO, H(2)O-CS(2), H(2)O-CO(2), and H(2)O-N(2)O are compared. The first two are linear and the others are T shaped with an O-C/O-N van der Waals bond, i.e., the oxygen of water bonds to the carbon and nitrogen of CO(2) and N(2)O, respectively. PMID- 15549862 TI - Dissociative electron attachment to dinitrogen pentoxide, N2O5. AB - Electron attachment was studied in gaseous dinitrogen pentoxide, N(2)O(5), for incident electron energies between a few meV and 10 eV. No stable parent anion N(2)O(5) (-) was observed but several anionic fragments (NO(3) (-), NO(2) (-), NO(-), O(-), and O(2) (-)) were detected using quadrupole mass spectrometry. Many of these dissociative pathways were found to be coupled and provide detailed information on the dynamics of N(2)O(5) fragmentation. Estimates of the cross sections for production of each of the anionic fragments were made and suggest that electron attachment to N(2)O(5) is amongst the most efficient attachment reactions recorded for nonhalogenated polyatomic systems. PMID- 15549863 TI - Different approaches for the calculation of electronic excited states of nonstoichiometric alkali halide clusters: the example of Na3F. AB - The electronic structure and excited states of the Na(3)F cluster are investigated using different approximate, but numerically efficient, computational schemes, such as a 2e hybrid quantum/classical pseudopotential model with full-configuration interaction or time-dependent density-functional theory. Various quantities such as geometries and transition energies are compared with results previously obtained by multireference configuration interaction calculations, taken as reference data. The potential energy surfaces of the lowest excited states are investigated and the finite-temperature absorption spectra are calculated. The good agreement with recent beam experiments [J.-M. L'Hermite, V. Blanchet, A. Le Padellec, B. Lamory, and P. Labastie, Eur. Phys. J. D 28, 361 (2004)] leads to the conclusion that the absorption spectrum observed experimentally corresponds to the lowest energy isomer which has a C(2v) planar rhombic geometry. PMID- 15549864 TI - Ultrafast excited state dynamics of the Na3F cluster: quantum wave packet and classical trajectory calculations compared to experimental results. AB - Short-time, excited-state dynamics of the lowest isomer of the Na(3)F cluster is studied theoretically in order to interpret the features of recent time-resolved pump-probe ionization experiments [J. M. L'Hermite, V. Blanchet, A. Le Padellec, B. Lamory, and P. Labastie, Eur. Phys. J. D 28, 361 (2004)]. In the present paper, we propose an identification of the vibrational motion responsible for the oscillations in the ion signal, on the basis of quantum mechanical wave packet propagations and classical trajectory calculations. The good agreement between experiment and theory allows for a clear interpretation of the detected dynamics. PMID- 15549865 TI - Magnitude and orientation dependence of intermolecular interaction of perfluoropropane dimer studied by high-level ab initio calculations: comparison with propane dimer. AB - Intermolecular interaction energies of 12 orientations of C(3)F(8) dimers were calculated with electron correlation correction by the second-order Moller Plesset perturbation method. The antiparallel C(2h) dimer has the largest interaction energy (-1.45 kcal/mol). Electron correlation correction increases the attraction considerably. Electrostatic energy is not large. Dispersion is mainly responsible for the attraction. Orientation dependence of the interaction energy of the C(3)F(8) dimer is substantially smaller than that of the C(3)H(8) dimer. The calculated interaction energy of the C(3)F(8) dimer at the potential minimum is 78% of that of the C(3)H(8) dimer (-1.85 kcal/mol), whereas the interaction energies of the CF(4) and C(2)F(6) dimers are larger than those of the CH(4) and C(2)H(6) dimers. The intermolecular separation in the C(3)F(8) dimer at the potential minimum is substantially larger than that in the C(3)H(8) dimer. The larger intermolecular separation due to the steric repulsion between fluorine atoms is the cause of the smaller interaction energy of the C(3)F(8) dimer at the potential minimum. The calculated intermolecular interaction energy potentials of the C(3)F(8) dimers using an all atom model OPLS-AA (OPLS all atom model) force field and a united atom model force field were compared with the ab initio calculations. Although the two force fields well reproduces the experimental vapor and liquid properties of perfluoroalkenes, the comparison shows that the united atom model underestimates the potential depth and orientation dependence of the interaction energy. The potentials obtained by the OPLS-AA force field are close to those obtained by the ab initio calculations. PMID- 15549868 TI - Optical-optical double resonance photoionization spectroscopy of nf Rydberg states of nitric oxide. AB - The spectra of vibrationally excited nf Rydberg states of nitric oxide were recorded by monitoring the photoion current produced using two-photon double resonance excitation via the NO A (2)Sigma(+) state followed by photoexcitation of the Rydberg state that undergoes autoionization. The optical transition intensities from NO A state to nf Rydberg states were calculated, and the results agree closely with experiment. These results combined with circular dichroism measurements allow us to assign rotational quantum numbers to the nf Rydberg states even in a spectrum of relatively low resolution. We report the positions of these nf (upsilon,N,N(c)) Rydberg levels converging to the NO X (1)Sigma(+) upsilon(+) = 1 and 2 ionization limits where N is the total angular momentum excluding electron and nuclear spin and N(c) represents the rotational quantum number of the ion core. Our two-color optical-optical double resonance measurements cover the range of N from 15 to 28, N(c) from 14 to 29, and the principal quantum number n from 9 to 21. The electrostatic interaction between the Rydberg electron and the ion core is used to account for the rotational fine structure and a corresponding model is used to fit the energy levels to obtain the quadrupole moment and polarizability of the NO(+) core. Comparison with a multichannel quantum defect theory fit to the same data confirms that the model we use for the electrostatic interaction between the nf Rydberg electron and the ion core of NO well describes the rotational fine structure. PMID- 15549869 TI - Collisional deexcitation of optically allowed excited atoms by axially symmetric molecules. AB - The deexcitation process of an atom in an optically allowed excited state by a collision with an axially symmetric molecule is considered. In order to make a precise comparison with experimental data that have recently been obtained, we extend previous work for a Penning ionization process [T. Watanabe and K. Katsuura, J. Chem. Phys. 47, 800 (1967)]. Using the straight-line trajectory impact parameter method, the probability of deexcitation in the incident atom is described by a discrete-continuum excitation transfer mechanism. The effects of the ionization yield eta in molecular target and of the molecular anisotropic property of the optical transition dipole are considered. The cross-section formula sigma is presented by a similar formula for Penning ionization of the atomic target by introducing a stereo factor C(lambda) as sigma = C(lambda)[e(4)mu(2)mu(E, perpendicular)/(4piepsilon(0))(2) (2)hv](2/5). Here, v is the relative velocity of the colliding system and lambda is given by the ratio lambda =(mu(E||)/mu(E perpendicular)), where mu, mu(E perpendicular), mu(E||) are the transition dipole moments of an excited atom, A-->A(*), those of a molecule at energy E for the perpendicular component and the parallel component with respect to molecular axis. Applications to He(*)(2(1)P)+H(2) (or D(2)), Ne(*)[2p(5)((2)P(1/2))3 s (1)P(1)]+ H(2) (or D(2)) systems and systems of the same projectiles on C(6)H(6), (or C(6)D(6)) molecules are made. The results for hydrogen molecules are compared with the experimental data. PMID- 15549870 TI - Theoretical determination of parity-violating vibrational frequency differences between the enantiomers of chiral molecules. AB - A perturbation treatment has been used to compute the leading first- and second order parity-violating corrections to the vibrational energy levels of a chiral molecule. Assuming the molecular equilibrium geometry as expansion point of both parity-violating and parity-conserving potential-energy surfaces, it is shown that these corrections, i.e., harmonic and anharmonic contributions, are of the same order of magnitude and that none of them can be neglected for a realistic determination of vibrational frequency differences. Numerical tests based on ab initio MP2 force fields and quantum-relativistic calculations of the parity violating potential for each normal mode of PHBrF and AsHBrF molecules confirm this conclusion. In particular, it is shown that a normal mode of AsHBrF is characterized by one of the largest vibrational frequency difference ever found- the prediction is approximately 0.1 Hz--only one order of magnitude less than the presumed resolution limit of current experimental investigations. PMID- 15549871 TI - Spatially resolved gas phase composition measurements in supersonic flows using tunable diode laser absorption spectroscopy. AB - We used a tunable diode laser absorption spectrometer to follow the condensation of D(2)O in a supersonic Laval nozzle. We measured both the concentration of the condensible vapor and the spectroscopic temperature as a function of position and compared the results to those inferred from static pressure measurements. Upstream and in the early stages of condensation, the quantitative agreement between the different experimental techniques is good. Far downstream, the spectroscopic results predict a lower gas phase concentration, a higher condensate mass fraction, and a higher temperature than the pressure measurements. The difference between the two measurement techniques is consistent with a slight compression of the boundary layers along the nozzle walls during condensation. PMID- 15549872 TI - Double proton transfer behavior and one-electron oxidation effect in double H bonded glycinamide-formic acid complex. AB - The behavior of double proton transfer occurring in a representative glycinamide formic acid complex has been investigated at the B3LYP/6-311 + + G( * *) level of theory. Thermodynamic and, especially, kinetic parameters, such as tautomeric energy, equilibrium constant, and barrier heights have been discussed, respectively. The relevant quantities involved in the double proton transfer process, such as geometrical changes, interaction energies, and intrinsic reaction coordinate calculations have also been studied. Computational results show that the participation of a formic acid molecule favors the proceeding of the proton transfer for glycinamide compared with that without mediate-assisted case. The double proton transfer process proceeds with a concerted mechanism rather than a stepwise one since no ion-pair complexes have been located during the proton transfer process. The calculated barrier heights are 11.48 and 0.85 kcal/mol for the forward and reverse directions, respectively. However, both of them have been reduced by 2.95 and 2.61 kcal/mol to 8.53 and -1.76 kcal/mol if further inclusion of zero-point vibrational energy corrections, where the negative barrier height implies that the reverse reaction should proceed with barrierless spontaneously, analogous to that occurring between glycinamide and formamide. Furthermore, solvent effects on the thermodynamic and kinetic processes have also been predicted qualitatively employing the isodensity surface polarized continuum model within the framework of the self-consistent reaction field theory. Additionally, the oxidation process for the double H-bonded glycinamide-formic acid complex has also been investigated. Contrary to that neutral form possessing a pair of two parallel intermolecular H bonds, only a single H bond with a comparable strength has been found in its ionized form. The vertical and adiabatic ionization potentials for the neutral complex have been determined to be about 9.40 and 8.69 eV, respectively, where ionization is mainly localized on the glycinamide fragment. Like that ionized glycinamide-formamide complex, the proton transfer in the ionized complex is characterized by a single well potential, implying that the proton initially attached to amide N4 in the glycinamide fragment cannot be transferred to carbonyl O13 in the formic acid fragment at the geometry of the optimized complex. PMID- 15549873 TI - Ab initio benchmark study for the oxidative addition of CH4 to Pd: importance of basis-set flexibility and polarization. AB - To obtain a state-of-the-art benchmark potential energy surface (PES) for the archetypal oxidative addition of the methane C-H bond to the palladium atom, we have explored this PES using a hierarchical series of ab initio methods (Hartree Fock, second-order Moller-Plesset perturbation theory, fourth-order Moller Plesset perturbation theory with single, double and quadruple excitations, coupled cluster theory with single and double excitations (CCSD), and with triple excitations treated perturbatively [CCSD(T)]) and hybrid density functional theory using the B3LYP functional, in combination with a hierarchical series of ten Gaussian-type basis sets, up to g polarization. Relativistic effects are taken into account either through a relativistic effective core potential for palladium or through a full four-component all-electron approach. Counterpoise corrected relative energies of stationary points are converged to within 0.1-0.2 kcal/mol as a function of the basis-set size. Our best estimate of kinetic and thermodynamic parameters is -8.1 (-8.3) kcal/mol for the formation of the reactant complex, 5.8 (3.1) kcal/mol for the activation energy relative to the separate reactants, and 0.8 (-1.2) kcal/mol for the reaction energy (zero-point vibrational energy-corrected values in parentheses). This agrees well with available experimental data. Our work highlights the importance of sufficient higher angular momentum polarization functions, f and g, for correctly describing metal-d-electron correlation and, thus, for obtaining reliable relative energies. We show that standard basis sets, such as LANL2DZ+1f for palladium, are not sufficiently polarized for this purpose and lead to erroneous CCSD(T) results. B3LYP is associated with smaller basis set superposition errors and shows faster convergence with basis-set size but yields relative energies (in particular, a reaction barrier) that are ca. 3.5 kcal/mol higher than the corresponding CCSD(T) values. PMID- 15549874 TI - Electron attachment and detachment: electron affinities of isomers of trifluoromethylbenzonitrile. AB - Rate constants for electron attachment to the three isomers of trifluoromethylbenzonitrile [(CF(3))(CN)C(6)H(4), or TFMBN] were measured over the temperature range of 303-463 K in a 133-Pa He buffer gas, using a flowing afterglow Langmuir-probe apparatus. At 303 K, the measured attachment rate constants are 9.0 x 10(-8) (o-TFMBN), 5.5 x 10(-8) (m-TFMBN), and 8.9 x 10(-8) cm(3) s(-1) (p-TFMBN), estimated accurate to +/-25%. The attachment process formed only the parent anion in all three cases. Thermal electron detachment was observed for all three anion isomers, and rate constants for this reverse process were also measured. From the attachment and detachment results, the electron affinities of the three isomers of TFMBN were determined to be 0.70(o-TFMBN), 0.67(m-TFMBN), and 0.83 eV (p-TFMBN), all +/-0.05 eV. G3(MP2) [Gaussian-3 calculations with reduced Moller-Plesset orders (MP2)] calculations were carried out for the neutrals and anions. Electron affinities derived from these calculations are in good agreement with the experimental values. PMID- 15549875 TI - Reaction kinetics of the addition of atomic sulfur to nitric oxide. AB - The reaction of S((3)P(J)) with NO ((2)Pi) in an Ar bath gas has been studied by the laser photolysis-resonance fluorescence technique over 300-810 K at pressures from 60 to 800 mbar. The observed second-order rate constants are close to the low-pressure limit. Fitting of Troe's formalism to experiment, with an estimated F(cent) = 0.78 exp(-T/7445) and k(infinity) given subsequently, yields k(0) = (6.2+/-0.6) x 10(-33) exp(+ (940+/-40)/T) cm(6) molecule(-2) s(-1). Error limits are +/-25%. A theoretical analysis of this value suggests that the average energy transferred during collisions between Ar and the excited intermediate is DeltaE = -360(-160) (+90) cm(-1). Over 300-800 K, the high-pressure limit is predicted to be k(infinity) = 2.2 x 10(-10) (T/300)(0.24) cm(3) molecule(-1) s(-1). Doublet and quartet adducts between S and NO were characterized via CBS-QB3 theory. The kinetic data can be rationalized with SNO ((2)A(')) as the major product, and an ab initio estimate of Delta(f)H(298) for SNO is 176+/-8 kJ mol(-1). PMID- 15549876 TI - Mechanism for the formation of gas-phase protonated alcohol-ether adducts by VUV laser ionization and density-functional calculations. AB - The neutral vapors above liquid alcohol/ether mixtures, (diethyl ether/methanol, diethyl ether/ethanol, tetrahydrofuran/methanol, and tetrahydrofuran/ethanol) were co-expanded with He in a supersonic jet, ionized with a 118-nm vacuum ultraviolet laser, and detected in a time-of-flight mass spectrometer. In each case, features attributed to protonated alcohol-ether dimers and protonated ether monomers were observed, as well as those ions obtained by ionizing neat alcohol or ether samples alone. Theoretical calculations, carried out to establish the energetics of the various possible reactions leading to the formation of the observed binary adducts, indicate that the most thermodynamically favorable pathway corresponds to the addition of a protonated alcohol monomer to neutral ether. PMID- 15549877 TI - Time-resolved relaxation dynamics of Hgn- (11 + |1>) at t = 0. We find that Re rho(10)(t) oscillates with high frequency and shows a fast damping. Relaxation time of the oscillation amplitude is estimated to be 5.1 ps for a certain configuration of the solution. Then, secular approximation often used in the Redfield theory is found to work well, at least, in the present system. Population relaxation time for the first excited state and pure dephasing time may also be calculated from the component of Re rho(10)(t) to be 7.9 and 7.5 ps, respectively. Further, the many-particle measurement for Re rho(10)(t) gives the relaxation rate about three times faster than the single-measurement above. This comes from the inhomogeneity of the solute environment. We also found the fast oscillation in the diagonal part of the calculated density matrix, Re rho(11)(t). This oscillation is generated only when the initial density matrix includes the coherence. PMID- 15549881 TI - X-ray absorption spectra of water within a plane-wave Car-Parrinello molecular dynamics framework. AB - We describe the implementation of a simple technique to simulate core-level spectra within the Car-Parrinello plane-waves molecular dynamics framework. The x ray absorption (XA) spectra are generated using the transition potential technique with the effect of the core hole included through a specifically developed pseudopotential for the core-excited atom. Despite the lack of 1s core orbitals in the pseudopotential treatment, the required transition moments are accurately calculated without reconstruction of the all-electron orbitals. The method is applied to the oxygen XA spectra of water in its various aggregation states, but it is transferable to any first-row element. The computed spectra are compared favorably with the results from all-electron cluster calculations, as well as with experimental data. The periodicity of the plane-wave technique improves the description of condensed phases. The molecular dynamics simulation enables in principle a proper treatment of thermal effects and dynamical averaging in complex systems. PMID- 15549882 TI - The decay of pair correlations in quantum hard-sphere fluids. AB - A study of the asymptotic decay of the pair radial correlations in the bare quantum hard-sphere (QHS) fluid and in the quantum hard-sphere Yukawa (QHSY) fluid is presented. The conditions explored are far from quantum exchange and are contained within the region (0.1or=26 degrees C. Increases in temperature result in dehydration in the ethylene oxide chain, which increases the hydrophobicity, and impart to the molecules a longer-chain-like character. As a result line tension increases with increasing temperature, which probably outweighs the dipole-dipole repulsions showing circular domains at higher temperatures. The PM-IRRAS measurement reveals that the nu(as)(CH(2)) mode moves to lower wave numbers indicating that the LE-LC (liquid expanded-liquid condensed) phase transition during the compression of the monolayer involves changes in the conformational order of the molecules with a preferential increase in the planner trans zigzag conformation of the hydrocarbon chains. The nu(as)(CH(2)) mode in the LC region of the isotherm shows a constant value around 2917.8 cm(-1) indicating a stable state of the monolayer with an almost all-trans conformation of the hydrocarbon chains. The downward band at 1124 cm(-1) assigned to the nu(as)(C-O-C) mode indicates that the corresponding transition dipole moment is oriented perpendicular to the water surface. PMID- 15549899 TI - CdS:Mn nanocrystals passivated by ZnS: synthesis and luminescent properties. AB - Synthesis and characterization of highly luminescent ZnS-passivated CdS:Mn (CdS:Mn/ZnS) core/shell structured nanocrystals are reported. Mn-doped CdS core nanocrystals are produced ranging from 1.5 to 2.3 nm in diameter with epitaxial ZnS shell of wider band gap via a reverse micelle process. UV irradiation stimulated photo-oxidation of the ZnS shell results in formation of sulfate (ZnSO(4)) as determined by x-ray photoelectron spectroscopy, which increases the photoluminescence emission intensity and subsequent photostability. Luminescent relaxation lifetime data present two different decay components, consisting of slow decay emission from the Mn center and a fast decay emission from a defect related center. The impact of the density of surface defect states upon the emission spectra is discussed. PMID- 15549900 TI - First-principles study of C adsorption, O adsorption, and CO dissociation on flat and stepped Ni surfaces. AB - The adsorption of atomic oxygen and carbon was studied with plane wave density functional theory on four Ni surfaces, Ni(110), Ni(111), Ni(210), and Ni(531). Various adsorption sites on these surfaces are examined in order to identify the most favorable adsorption site for each atomic species. The dependence of surface bonding on adsorbate coverage is also investigated. Adsorption energies and structural information are obtained and compared with existing experimental results for Ni(110) and Ni(111). In addition, activation barriers to CO dissociation have been determined on Ni(111) and Ni(531) by locating the transition states for these processes. Our results indicate that the binding energies of C are comparatively stronger on stepped surfaces than on flat surfaces, and the energy barriers associated with CO dissociation strongly favor reactions occurring near surface steps. PMID- 15549901 TI - Solution combustion derived nanocrystalline Zn(2)SiO(4):Mn phosphors: a spectroscopic view. AB - Manganese doped nanocrystalline willemite powder phosphors Zn(2-x)Mn(x)SiO(4) (0.1(6)A(1) ground state. The mechanism involved in the generation of a green emission has been explained in detail. The effect of Mn content on luminescence has also been studied. PMID- 15549902 TI - Line shapes and satellites in high-resolution x-ray photoelectron spectra of large pi-conjugated organic molecules. AB - We present a high-resolution C1s and O1 s x-ray photoemission (XPS) study for condensed films of pi-conjugated organic molecules, namely, of the anhydrides 3,4,9,10-perylene-tetracarboxylic acid dianhydride, 1,4,5,8-naphthalene tetracarboxylic acid dianhydride, 1,8-naphthalene dicarboxylic acid anhydride, and benzoperylene-(1,8)-dicarboxylic acid anhydride as well as the quinoic acenaphthenequinone. Although the functional groups are identical for the anhydrides, the molecules show very different photoemission fine structure thus providing a detailed fingerprint. A simultaneous peak fit analysis of the XPS spectra of all molecules allows to consistently determine the ionization potentials of all chemically different carbon and oxygen atoms. Additional structures in the C1s and O1s spectra are interpreted as shakeup satellites and assigned with the help of singles and doubles configuration interaction calculations. These satellites provide further information on multielectron excitations and must be taken into account for quantitative investigations. PMID- 15549903 TI - Translocation of a beta-hairpin-forming peptide through a cylindrical tunnel. AB - We use Langevin dynamics simulations of a minimalist off-lattice model to study the translocation of a beta hairpin forming peptide through a tunnel that mimics the exit tunnel in a ribosome. We have computed the free energy of the peptide as a function of its position relative to the tunnel exit and also studied the properties of the conformational ensemble, when the peptide's position is restricted at different points along the tunnel. Confining the peptide within a sufficiently wide tunnel stabilizes the folded state. The protein then remains folded as it moves towards the tunnel exit. However, when the diameter D of the tunnel is below a certain critical value D(c), confinement destabilizes the folded state and forces the peptide to assume an extended configuration. In this case, as the peptide progresses towards the tunnel exit and eventually leaves the tunnel, it goes through a series of compact, misfolded conformations and eventually folds when it gets close to the exit. The critical tunnel diameter D(c) is comparable to the width of ribosomal tunnels. Our results suggest that co translational folding is probably not universal, but rather a protein-specific phenomenon. PMID- 15549904 TI - Fractional Brownian dynamics in proteins. AB - Correlation functions describing relaxation processes in proteins and other complex molecular systems are known to exhibit a nonexponential decay. The simulation study presented here shows that fractional Brownian dynamics is a good model for the internal dynamics of a lysozyme molecule in solution. We show that both the dynamic structure factor and the associated memory function fit well the corresponding analytical functions calculated from the model. The numerical analysis is based on autoregressive modeling of time series. PMID- 15549905 TI - Effect of salt bridges on the energy landscape of a model protein. AB - The effect of introducing salt bridges (gatekeepers) into an off-lattice three color, 46-bead model protein is investigated in terms of the effect on global optimization statistics. The global minima for all the gatekeepers that exhibited faster folding in previous molecular dynamics studies are located more rapidly than for the original potential, although the global minimum itself may change. Visualization of the underlying potential energy surface using disconnectivity graphs reveals that the gatekeepers exhibit structure intermediate between the original potential and a Go model. Competition between low-lying minima and the global minimum is reduced in the gatekeepers compared to the original potential, and interconversion barriers are generally smaller. PMID- 15549906 TI - General relationships between the mobility of a chain fluid and various computed scalar metrics. AB - We performed molecular dynamics simulations of chain systems to investigate general relationships between the system mobility and computed scalar quantities. Three quantities were found that had a simple one-to-one relationship with mobility: packing fraction, potential energy density, and the value of the static structure factor at the first peak. The chain center-of-mass mobility as a function of these three quantities could be described equally well by either a Vogel-Fulcher type or a power law equation. PMID- 15549907 TI - Discrimination in isotropic, nematic, and smectic phases of chiral calamitic molecules: a computer simulation study. AB - Racemic fluids of chiral calamitic molecules are investigated with molecular dynamics simulations. In particular, the phase behavior as a function of density is examined for eight racemates. The relationship between chiral discrimination and orientational order in the phase is explored. We find that the transition from the isotropic phase to a liquid crystal phase is accompanied by an increase in chiral discrimination, as measured by differences in radial distributions. Among ordered phases, discrimination is largest for smectic phases with a significant preference for heterochiral contact within the layers. PMID- 15549908 TI - The chemical activity of metal compound nanoparticles: importance of electronic and steric effects in M8C12 (M=Ti, V, Mo) metcars. AB - Density functional theory was employed to investigate the chemical activity of metal carbide nanoparticles. The present calculations indicate that M(8)C(12) (M=Ti, V, Mo) nanoparticles exhibit a unique behavior compared to metal [M(001)] and metal carbide surfaces [M(2)C(001) and MC(001)]. It is found that the nanoparticles behave very reactive in spite of the high carbon concentration in some reactions, while surprisingly inert in other cases. Our study reveals that the unexpected activity is the result of the interplay of shifts in the metal d bands and distortions in the geometry of the metal carbide nanoparticles. PMID- 15549909 TI - Solid state protein monolayers: morphological, conformational, and functional properties. AB - We have studied the morphological, conformational, and electron-transfer (ET) function of the metalloprotein azurin in the solid state, by a combination of physical investigation methods, namely atomic force microscopy, intrinsic fluorescence spectroscopy, and scanning tunneling microscopy. We demonstrate that a "solid state protein film" maintains its nativelike conformation and ET function, even after removal of the aqueous solvent. PMID- 15549910 TI - Interpolating moving least-squares methods for fitting potential energy surfaces: Analysis of an application to a six-dimensional system. AB - The basic formal and numerical aspects of different degree interpolated moving least-squares (IMLS) methods are applied to a six-dimensional potential energy surface (PES) of the HOOH molecule, for which an analytic ("exact") potential is available in the literature. The results of systematic investigations of the effects of weight function parameters, the degree and partial degree of IMLS, the number of data points allowed, and the optimal automatic point selection of data points up to full third-degree IMLS fits are reported. With partial reduction of cross terms and automatic point selection the full six-dimensional HOOH PES can be fit over a range of 100 kcal/mol to an accuracy of less than 1 kcal/mol with approximately 1350 ab initio points. PMID- 15549911 TI - Functional dependence of core-excitation energies. AB - We examine in depth the functional dependence of computed core-electron binding and excitation energies based on a total-energy difference approach within Kohn Sham density functional theory. Twenty-seven functional combinations were studied using a database of reliable experimental data on 18 molecules. The computed core electron binding energies are largely dependent on the choice of exchange functional. The term value of the first resonant excited state and energy differences between the lowest core-excited states are, however, quite insensitive to the choice of functionals since the errors due to the core-region cancel out. Using these results we define a different exchange functional, which mixes two functionals designed by Perdew and Wang (PD86 and PD91), with the best results for both excitation and binding energies obtained for a mixing ratio 60:40 between these. We also reexamine the relativistic corrections for inner shell excitations. PMID- 15549912 TI - High-order averaging schemes with error bounds for thermodynamical properties calculations by molecular dynamics simulations. AB - We introduce high-order formulas for the computation of statistical averages based on the long-time simulation of molecular dynamics trajectories. In some cases, this allows us to significantly improve the convergence rate of time averages toward ensemble averages. We provide some numerical examples that show the efficiency of our scheme. When trajectories are approximated using symplectic integration schemes (such as velocity Verlet), we give some error bounds that allow one to fix the parameters of the computation in order to reach a given desired accuracy in the most efficient manner. PMID- 15549913 TI - Binomial leap methods for simulating stochastic chemical kinetics. AB - This paper discusses efficient simulation methods for stochastic chemical kinetics. Based on the tau-leap and midpoint tau-leap methods of Gillespie [D. T. Gillespie, J. Chem. Phys. 115, 1716 (2001)], binomial random variables are used in these leap methods rather than Poisson random variables. The motivation for this approach is to improve the efficiency of the Poisson leap methods by using larger stepsizes. Unlike Poisson random variables whose range of sample values is from zero to infinity, binomial random variables have a finite range of sample values. This probabilistic property has been used to restrict possible reaction numbers and to avoid negative molecular numbers in stochastic simulations when larger stepsize is used. In this approach a binomial random variable is defined for a single reaction channel in order to keep the reaction number of this channel below the numbers of molecules that undergo this reaction channel. A sampling technique is also designed for the total reaction number of a reactant species that undergoes two or more reaction channels. Samples for the total reaction number are not greater than the molecular number of this species. In addition, probability properties of the binomial random variables provide stepsize conditions for restricting reaction numbers in a chosen time interval. These stepsize conditions are important properties of robust leap control strategies. Numerical results indicate that the proposed binomial leap methods can be applied to a wide range of chemical reaction systems with very good accuracy and significant improvement on efficiency over existing approaches. PMID- 15549914 TI - The accuracy of diffusion quantum Monte Carlo simulations in the determination of molecular equilibrium structures. AB - For a test set of 17 first-row small molecules, the equilibrium structures are calculated with Ornstein-Uhlenbeck diffusion quantum Monte Carlo simulations guiding by trial wave functions constructed from floating spherical Gaussian orbitals and spherical Gaussian geminals. To measure performance of the Monte Carlo calculations, the mean deviation, the mean absolute deviation, the maximum absolute deviation, and the standard deviation of Monte Carlo calculated equilibrium structures with respect to empirical equilibrium structures are given. This approach is found to yield results having a uniformly high quality, being consistent with empirical equilibrium structures and surpassing calculated values from the coupled cluster model with single, double, and noniterative triple excitations [CCSD(T)] with the basis sets of cc-pCVQZ and cc-pVQZ. The nonrelativistic equilibrium atomization energies are also presented to assess performance of the calculated methods. The mean absolute deviations regarding experimental atomization energy are 0.16 and 0.21 kcal/mol for the Monte Carlo and CCSD(T)/cc-pCV(56)Z calculations, respectively. PMID- 15549915 TI - Combinatorial invariants and covariants as tools for conical intersections. AB - The combinatorial invariant and covariant are introduced as practical tools for analysis of conical intersections in molecules. The combinatorial invariant is a quantity depending on adiabatic electronic states taken at discrete nuclear configuration points. It is invariant to the phase choice (gauge) of these states. In the limit that the points trace a loop in nuclear configuration space, the value of the invariant approaches the corresponding Berry phase factor. The Berry phase indicates the presence of an odd or even number of conical intersections on surfaces bounded by these loops. Based on the combinatorial invariant, we develop a computationally simple and efficient method for locating conical intersections. The method is robust due to its use of gauge invariant nature. It does not rely on the landscape of intersecting potential energy surfaces nor does it require the computation of nonadiabatic couplings. We generalize the concept to open paths and combinatorial covariants for higher dimensions obtaining a technique for the construction of the gauge-covariant adiabatic-diabatic transformation matrix. This too does not make use of nonadiabatic couplings. The importance of using gauge-covariant expressions is underlined throughout. These techniques can be readily implemented by standard quantum chemistry codes. PMID- 15549916 TI - Theoretical study of the electronic properties of narrow single-walled carbon nanotubes: beyond the local density approximation. AB - In this work we present a systematic density functional theory study of the electronic properties of single-walled carbon nanotubes (SWNT) with diameters ranging from 3 to 5 A. In this work meta-generalized-gradient approximation, hybrid, and screened exchange hybrid functionals are utilized to compute energy band gaps in these narrow SWNT. Our calculations using hybrid functionals show that the only true exceptions to the zone folding predictions are the (4,0) and (5,0) SWNT. The remaining chiral SWNT are semiconducting with band gaps that can be as large as 1.7 eV. However, the calculated energy band gaps are significantly smaller than those predicted by the zone folding scheme. This difference is primarily attributed to the sigma-pi hybridization present in such narrow SWNT. PMID- 15549917 TI - Atom distributions in binary atom clusters: a perturbational approach and its validation in a case study. AB - An approach to describe heteroatomic clusters A(n)B(N-n) as perturbed homoatomic ones is presented. By first treating the homoatomic systems A(N) or/and B(N) and subsequent application of first-order perturbation theory it is possible to estimate relative stabilities of the 2(N) possible distributions of the atom types A and B at the N atomic sites in a very efficient manner. The approach was tested considering Ir(n)Pt(13-n) as an example (treated with density functional methods). One observes good correlation between relative stabilities estimated from the homoatomic cases and those obtained from explicit treatments of the binary systems. Moreover we rationalize the observed correlation of atom type and atom position in Ir(n)Pt(13-n). PMID- 15549918 TI - A new localization scheme for the elongation method. AB - A different localization scheme for the elongation method is developed based on regional molecular orbitals. This scheme is more efficient and more accurate than the previous one especially for covalently bonded systems with strongly delocalized pi electrons. Ab initio test calculations have been performed on three model systems: water chains, polyglycine, and cationic cyanine chains. The dependence on the size of the starting clusters and the effect of the basis set are investigated. Our results are compared with conventional ab initio calculations and it is found in all cases that the error per added unit levels off to a satisfactorily small value as long as the starting cluster is sufficiently large. PMID- 15549919 TI - Equilibrium free energy estimates based on nonequilibrium work relations and extended dynamics. AB - Jarzynski's relation and the fluctuation theorem have established important connections between nonequilibrium statistical mechanics and equilibrium thermodynamics. In particular, an exact relationship between the equilibrium free energy and the nonequilibrium work is useful for computer simulations. In this paper, we exploit the fact that the free energy is a state function, independent of the pathway taken to change the equilibrium ensemble. We show that a generalized expression is advantageous for computer simulations of free energy differences. Several methods based on this idea are proposed. The accuracy and efficiency of the proposed methods are evaluated with a model problem. PMID- 15549920 TI - Pulse-pair control of resonance leaking in molecular multiphoton transitions. AB - We use model five-level systems to study resonance leaking of pi-pulse-induced multiphoton (MP) transitions along a strongly coupled anharmonic ladder. We demonstrate that the presence of a weakly bound background state attached to the ladder either in linear or Lambda configuration can have very pronounced effects on resonant MP ladder transitions, including essentially complete quenching of the primary transition. We also develop control strategies for the elimination of background state population based on phase-adjusted Gaussian pulse pairs and discuss the underlying control mechanisms. Finally we show that these strategies are effective in realistic molecular many-level systems. In particular, we demonstrate efficient pulse-pair control of resonance leaking in a 165-level system modeling vibrational excitation in HCN. PMID- 15549921 TI - Dipole-quadrupole and dipole-octopole polarizability of OsO4 from depolarized collision-induced light scattering experiments, ab initio and density functional theory calculations. AB - The dipole-quadrupole and dipole-octopole polarizability of osmium tetroxide (OsO(4)) has been determined from collision-induced light-scattering experiments. Our final estimates for these properties are |A|=(84+/-5)e(2)a(3)(0)E(-1)(h) and |E|=(214+/-25)e(2)a(4)(0)E(-1)(h). We have also analyzed previous experimental data of the relative permittivity and refractivity of OsO(4) to propose the electronic part of the static dipole polarizability of alpha=51.0e(2)a(2)(0)E( 1)(h). To support our findings we have performed high-level ab initio and density functional theory calculations to obtain theoretical static estimates alpha=(50.2+/-1.6)e(2)a(2)(0)E(-1)(h), A=(84+/-10)e(2)a(3)(0)E(-1)(h), and E=( 252+/-32)e(2)a(4)(0)E(-1)(h), in essential agreement with the proposed experimental values. PMID- 15549922 TI - Accurate intermolecular ground state potential of the Ar-N2 van der Waals complex. AB - After carrying out a systematic basis set convergence study, we evaluate several ground state potential energy surfaces of the Ar-N(2) van der Waals complex at the coupled cluster singles and doubles model including connected triples corrections. We use the aug-cc-pVXZ (X=5,Q,D) and the daug-cc-pVQZ basis sets augmented with a set of 3s3p2d1f1g (denoted 33211) and 3s3p2d2f1g (denoted 33221) midbond functions, respectively. aug-cc-pVTZ-33211 results were available in the literature. The aug-cc-pV5Z-33211 (daug-cc-pVQZ-33221) surface is characterized by a T-shaped minimum at R(e)=3.709 (3.701) A and of 99.01 (102.50) cm(-1), and a linear saddle point at 4.260 (4.257) A and D(e)=75.28 (79.73) cm(-1). These results are compared with the values provided by the semiempirical potentials available, and those of previous theoretical studies. The basis set convergence of the intermolecular potentials is also analyzed. From the potentials the rovibronic spectroscopic properties are determined. We study the basis set convergence of the rotational frequencies. The binding parameters that characterized the aug-cc-pVTZ-33211 surface are reasonable, but the surface is not good enough to evaluate the microwave spectra. The aug-cc-pVQZ-33211 basis set results considerably improve the triple zeta and are close to the aug-cc-pV5Z 33211. Considering the small differences between the quadruple and the quintuple zeta surfaces, the latter results can be expected to be close to convergence. At this level the differences with respect to the accurate experimental frequencies are in the order of 0.7%. In the case of the daug-cc-pVXZ-33211,33221 (X=5,Q,T,D) series, the convergence of the interaction energies with respect to basis set improvement is not so smooth. The errors in the frequencies obtained with the daug-cc-pVQZ-33221 basis set with respect to experiment are in the order of 0.4%. PMID- 15549923 TI - Product spin-orbit state resolved dynamics of the H+H2O and H+D2O abstraction reactions. AB - The product state-resolved dynamics of the reactions H+H(2)O/D(2)O- >OH/OD((2)Pi(Omega);v',N',f )+H(2)/HD have been explored at center-of-mass collision energies around 1.2, 1.4, and 2.5 eV. The experiments employ pulsed laser photolysis coupled with polarized Doppler-resolved laser induced fluorescence detection of the OH/OD radical products. The populations in the OH spin-orbit states at a collision energy of 1.2 eV have been determined for the H+H(2)O reaction, and for low rotational levels they are shown to deviate from the statistical limit. For the H+D(2)O reaction at the highest collision energy studied the OD((2)Pi(3/2),v'=0,N'=1,A') angular distributions show scattering over a wide range of angles with a preference towards the forward direction. The kinetic energy release distributions obtained at 2.5 eV also indicate that the HD coproducts are born with significantly more internal excitation than at 1.4 eV. The OD((2)Pi(3/2),v'=0,N'=1,A') angular and kinetic energy release distributions are almost identical to those of their spin-orbit excited OD((2)Pi(1/2),v'=0,N'=1,A') counterpart. The data are compared with previous experimental measurements at similar collision energies, and with the results of previously published quasiclassical trajectory and quantum mechanical calculations employing the most recently developed potential energy surface. Product OH/OD spin-orbit effects in the reaction are discussed with reference to simple models. PMID- 15549924 TI - Quantum yields for product formation in the 120-133 nm photodissociation of O2. AB - The photodissociation of O(2) in the region from 120-133 nm has been investigated using product imaging. The spectrum in this region is dominated by transitions from the ground state to the first three vibrational levels of the E (3)Sigma(u) (-) state. The O((1)D)+O((3)P) channel is the only product channel observed by product imaging for dissociation at either 124.4 nm or 120.4 nm. The O((1)D(2)) product is aligned in the molecular frame in such a way that its J vector is perpendicular to the relative velocity vector between the O((1)D) and the O((3)P). The variation in the anisotropy of dissociation is approximately predicted by considering transitions on individual lines and then taking into account the coherent excitation of overlapping resonances. At 132.7 nm, both the O((1)D)+O((3)P) and the O((3)P)+O((3)P) channels are observed with branching ratios of 0.40+/-0.08 and 0.60+/-0.09, respectively. At 130.2 nm, the quantum yield for production of O((1)D) is 0.76+/-0.28. PMID- 15549925 TI - Theoretical study of the electronic structure of CnS (n=1-6) thiocumulenes. AB - Linear sulfur-carbon chains C(n)S (n=1-6) of astronomical interest were examined by means of several theoretical methods. The three smallest compounds of the series were chosen to evaluate the performance of several computational models, including Hartree-Fock theory, density functional theory with the Becke's three parameter exchange functional and the correlation functional of Lee, Yang, and Parr (B3LYP), and electron-correlated methods (second-order Moller-Plesset perturbation method (MP2), configuration interaction method including single and double excitations (CISD), and quadratic configuration interaction method including single and double excitations (QCISD) in combination with a large variety of basis sets. The systematic comparison between the experiment and theory indicates that the B3LYP/6-311G** method can be considered suitable for the study of the electronic structures of the C(n)S compounds. The electronic ground states of the C(n)S molecules alternate between 1Sigma and 3Sigma for odd and even values of n, respectively. The B3LYP/6-311G** wave functions for these electronic ground states were analyzed by means of the atoms in molecules (AIM) and natural bond orbital (NBO) methods. Both approaches suggest that the electronic structures for the singlet and triplet compounds must be considered separately. According to the NBO method, singlet compounds can be properly represented by acetylenic structures with alternating single and triple bonds (S[triple bond]C-C[triple bond]C...). However, triplet compounds are better described by means of double bond-double bond cumulenic structures (S=C=C=C=C...) as a consequence of the average between different alpha and beta electronic densities. AIM delocalization indexes and NBO interactions between localized orbitals also indicate that these structures are strongly pi delocalized. Finally, the different singlet and triplet structures proposed provide a consistent explanation for the geometries, dipole moments, and spin-density values of the C(n)S compounds studied. PMID- 15549926 TI - Formation and properties of halogenated aluminum clusters. AB - The fast-flow tube reaction apparatus was employed to study the halogenation of aluminum clusters. For reactions with HX (X=Cl, Br, and I), acid-etching pathways are evident, and we present findings for several reactions, whereby Al(n)X(-) generation is energetically favorable. Tandem reaction experiments allowed us to establish that for Al(n)Cl(-), Al(n)I(-), and Al(n)I(2) (-), species with n=6, 7, and 15 are particularly resistant to attack by oxygen. Further, trends in reactivity suggest that, in general, iodine incorporation leaves the aluminum clusters' electronic properties largely unperturbed. Ab initio calculations were performed to better interpret reaction mechanisms and elucidate the characteristics of the products. Lowest energy structures for Al(13)X(-) were found to feature icosahedral Al(13) units with the halogen atom located at the on top site. The charge density of the highest occupied molecular orbital in these clusters is heavily dependent on the identity of X. The dependence of reactivity on the clusters' charge state is also discussed. In addition, we address the enhanced stability of Al(13)I(-) and Al(13)I(2) (-), arguing that the superhalogen behavior of Al(13) in these clusters can provide unique opportunities for the synthesis of novel materials with saltlike structures. PMID- 15549927 TI - The nature of the complex formed between pyridine and hydrogen bromide in the gas phase: an experimental approach using rotational spectroscopy. AB - The rotational spectra of two isotopomers, C(5)H(5)N...H(79)Br and C(5)H(5)N...H(81)Br, of a complex formed by pyridine and hydrogen bromide were observed by using a pulsed-jet, Fourier-transform microwave spectrometer that incorporated a mixing nozzle. Rotational constants, centrifugal distortion constants, and nuclear quadrupole coupling constants for the (14)N and Br nuclei were determined. The rotational constants indicate that the observed complex has a planar geometry of C(2v) symmetry, with the HBr subunit lying along the C(2) axis of pyridine. These conclusions are in good agreement with those obtained earlier from infrared spectroscopy in cryogenic matrices and ab initio calculations. The distance r(N...H)=1.137(2) A was obtained by fitting the rotational constants under the assumption of an unchanged pyridine geometry and with r(H-Br) fixed at the value obtained through an ab initio calculation. The nuclear quadrupole coupling constants were interpreted, with the aid of ab initio calculations for the complex, as well as hydrogen-bonded and ion-pair limiting models for the interaction of pyridine and HBr, to establish that the complex may be visualized in terms of a substantial contribution (>60%) of the ionic structure C(5)H(5)NH(+)...Br(-) to its valence-bond description. The intermolecular force constant k(sigma)=57.8(16) N m(-1) determined from the centrifugal distortion constants Delta(J) of the two isotopomers is also consistent with a strongly bound complex involving partial proton transfer. PMID- 15549928 TI - Reactions of silicon atoms and small clusters with CO: experimental and theoretical characterization of SinCO (n=1-5), Si2(CO)2, c-Si2(mu-O)(mu-CSi), and c-Si2(mu-O)(mu-CCO) in solid argon. AB - Reactions of silicon atoms and small clusters with carbon monoxide molecules in solid argon have been studied using matrix isolation infrared absorption spectroscopy. In addition to the previously reported SiCO monocarbonyl, Si(2)(CO)(2) and Si(n)CO (n=2-5) carbonyl molecules were formed spontaneously on annealing and were characterized on the basis of isotopic substitution and theoretical calculations. It was found that Si(2)CO, Si(3)CO, and Si(5)CO are bridge-bonded carbonyl compounds, whereas Si(4)CO is a terminal-bonded carbonyl molecule. The Si(2)(CO)(2) and Si(3)CO molecules photochemically rearranged to the more stable c-Si(2)(mu-O)(mu-CCO) and c-Si(2)(mu-O) (mu-CSi) isomers where Si(2) is inserted into the CO triple bond. PMID- 15549929 TI - Dissociative recombination study of Na+ (D2O) in a storage ring. AB - The dissociative recombination of Na(+)(D(2)O) ion has been studied at the heavy ion storage ring CRYRING (Manne Siegbahn Laboratory, Stockholm University). The cross section has been measured as a function of center-of-mass energy ranging from 1 meV to 0.1 eV and found to have an E(-1.37) dependence. The rate coefficient has been deduced to be (2.3+/-0.32)x10(-7)(T(e)/300)(-0.95+/-0.01) cm(3) s(-1) for T(e)=50-1000 K. The branching ratios have been measured at 0 eV. Of the four energetically accessible dissociation channels, three channels are found to occur although the channel that breaks the weak Na(+)-D(2)O bond is by far dominant. PMID- 15549930 TI - The static polarizability and first hyperpolarizability of the water trimer anion: ab initio study. AB - This work predicts the extraordinary hyperpolarizability of inorganic clusters: two water trimer anions. The first hyperpolarizabilities of (H2O-)(3) are considerable, beta(0)=1.715 x 10(7) a.u. for configuration A and beta(0)=1.129 x 10(7) a.u. for configuration B at MP2/d-aug-cc-pVDZ+x level. The first hyperpolarizabilities of (H2O-)(3) (configuration A) and related systems [(H2O)(3) and (H2O)(3)F-] are compared at the MP2/d-aug-cc-pVDZ+x level. These results are beta(0)=1.715 x 10(7) a.u. for (H2O-)(3), beta(0)=35 a.u. for (H2O)(3) [the neutral core of (H2O-)(3)], and beta(0)=46 a.u. for (H2O)(3)F-). Comparing the beta(0) values of related systems, we find that the dipole-bound excess electron is the key factor in the extraordinary first hyperpolarizability of (H2O-)(3) species. It will provide a future in the development of some materials with the excess electron (e.g., electrides) that exhibit large nonlinear optical response. PMID- 15549931 TI - Electron affinities with diffusion quantum Monte Carlo for C2 and BO molecules. AB - To verify the performance of the fixed-node diffusion quantum Monte Carlo method in electron affinities calculations, the adiabatic electron affinities of C(2) and BO molecules calculated by the fixed-node Ornstein-Uhlenbeck diffusion quantum Monte Carlo simulations guiding by trial wave functions constructed from the floating spherical Gaussian orbitals and spherical Gaussian geminals are presented in this work. The random walk based results, 3.264(43) and 2.507(32) eV for C(2) and BO, respectively, are compared with the available best experimental determined values, 3.269(6) and 2.508(8) eV for C(2) and BO, respectively, and the results of other theoretical calculations. PMID- 15549932 TI - Growth dynamics and intracluster reactions in Ni+(CO2)n complexes via infrared spectroscopy. AB - Ni(+)(CO(2))(n), Ni(+)(CO(2))(n)Ar, Ni(+)(CO(2))(n)Ne, and Ni(+)(O(2))(CO(2))(n) complexes are generated by laser vaporization in a pulsed supersonic expansion. The complexes are mass-selected in a reflectron time-of-flight mass spectrometer and studied by infrared resonance-enhanced photodissociation (IR-REPD) spectroscopy. Photofragmentation proceeds exclusively through the loss of intact CO(2) molecules from Ni(+)(CO(2))(n) and Ni(+)(O(2))(CO(2))(n) complexes, and by elimination of the noble gas atom from Ni(+)(CO(2))(n)Ar and Ni(+)(CO(2))(n)Ne. Vibrational resonances are identified and assigned in the region of the asymmetric stretch of CO(2). Small complexes have resonances that are blueshifted from the asymmetric stretch of free CO(2), consistent with structures having linear Ni(+)-O=C=O configurations. Fragmentation of larger Ni(+)(CO(2))(n) clusters terminates at the size of n=4, and new vibrational bands assigned to external ligands are observed for n> or =5. These combined observations indicate that the coordination number for CO(2) molecules around Ni(+) is exactly four. Trends in the loss channels and spectra of Ni(+)(O(2))(CO(2))(n) clusters suggest that each oxygen atom occupies a different coordination site around a four coordinate metal ion in these complexes. The spectra of larger Ni(+)(CO(2))(n) clusters provide evidence for an intracluster insertion reaction assisted by solvation, producing a metal oxide-carbonyl species as the reaction product. PMID- 15549933 TI - The double photoionization of HCl: an ion-electron coincidence study. AB - The double photoionization of HCl molecules by synchrotron radiation has been studied in the energy range between 30 and 50 eV. The HCl(2+) and Cl(2+) product ions have been detected by a photoelectron-photoion-coincidence technique, while the H(+)+Cl(+) formation, which follows the double ionization of HCl, has been studied by photoelectron-photoion-photoion coincidence. The photon energy threshold for the production of HCl(2+) ions has been found to be 35.4+/-0.6 eV, while for the dissociative channel leading to H(+)+Cl(+), it has been measured a threshold at 36.4+/-0.6 eV and a change in the slope of the cross-section energy dependence at 38.7+/-0.7 eV. The production of H+Cl(2+) occurs with a threshold photon energy of 42.8+/-1.1 eV. These results appear to be in a good agreement with previous data by different experimental techniques and recent theoretical calculations performed by our laboratory. PMID- 15549934 TI - Doubly excited 2 1Delta g state of Na2. AB - The doubly excited valence (3p+3p) 2 (1)Delta(g) state of Na(2) is experimentally observed by using optical-optical double resonance spectroscopy. A single line Ar(+) laser (a total of nine lines) was used to pump the sodium dimers from thermally populated ground state X (1)Sigma(g) (+) to the intermediate B (1)Pi(u) state. Then, a single mode Ti:sapphire laser was used to probe the doubly excited 2 (1)Delta(g) state. Violet fluorescence emitted from the highly excited states (mainly 2 (3)Pi(g) or 3 (3)Pi(g) states which are transferred from 2 (1)Delta(g) state via collision) to the a (3)Sigma(u) (+) state was monitored by a filtered photomultiplier tube (PMT). A total of 582 rovibrational levels of 2 (1)Delta(g) state were observed, identified, and assigned to the vibrational and rotational quantum numbers in the range of 0< or =v< or =28 and 11< or =J< or =99, respectively. The absolute vibrational quantum number assignment was verified by comparing the totally resolved fluorescence with the calculated Franck-Condon factors between 2 (1)Delta(g) state and B (1)Pi(u) state. Dunham coefficients and Rydberg-Klein-Rees potential curve were derived from these observed quantum levels. The primary molecular constants of Na(2) 2 (1)Delta(g) state are T(e)=32 416.759(15) cm(-1), omega(e)=124.8484(36) cm(-1), B(e)=0.119 158(3) cm(-1), and R(e)=3.508 20(5) A. PMID- 15549935 TI - Bonding and (hyper)polarizability in the sodium dimer. AB - We report a conventional ab initio and density functional theory study of the polarizability (alpha(alphabeta)/e(2)a(0) (2)E(h) (-1)) and hyperpolarizability (gamma(alphabetagammadelta)/e(4)a(0) (4)E(h) (-3)) of the sodium dimer. A large [18s14p9d2f1g] basis set is thought to yield near-Hartree-Fock values for both properties: alpha=272.28, Deltaalpha=127.22 and gamma=2157.6 x 10(3) at R(e)=3.078 87 A. Electron correlation has a remarkable effect on the Cartesian components of gamma(alphabetagammadelta). Our best value for the mean is gamma=1460.1 x 10(3). The (hyper)polarizability shows very strong bond-length dependence. The effect is drastically different for the longitudinal and transverse components of the hyperpolarizability. The following first derivatives were extracted from high-level coupled cluster calculations: (dalpha/dR)(e)=54.1, (dDeltaalpha/dR)(e)=88.1e(2)a(0)E(h) (-1), and (dgamma/dR)(e)=210 x 10(3)e(4)a(0) (3)E(h) (-3). We associate the (hyper)polarizability to bonding effects between the two sodium atoms by introducing the differential property per atom Q(diff)/2 identical with (Q[Na(2)(X (1)Sigma(g) (+))]/2-Q[Na((2)S)]). The differential (hyper)polarizability per atom is predicted to be strongly negative for the dimer at R(e), as [alpha(Na(2))/2-alpha(Na)]=-33.8 and [gamma(Na(2))/2-gamma(Na)]= 226.3 x 10(3). The properties calculated with the widely used B3LYP and B3PW91 density functional methods differ significantly. The B3PW91 results are in reasonable agreement with the conventional ab initio values. Last, we observe that low-level ab initio and density functional theory methods underestimate the dipole polarizability anisotropy. Experimental data on this important property are highly desirable. PMID- 15549936 TI - Norbornane: an investigation into its valence electronic structure using electron momentum spectroscopy, and density functional and Green's function theories. AB - We report on the results of an exhaustive study of the valence electronic structure of norbornane (C(7)H(12)), up to binding energies of 29 eV. Experimental electron momentum spectroscopy and theoretical Green's function and density functional theory approaches were all utilized in this investigation. A stringent comparison between the electron momentum spectroscopy and theoretical orbital momentum distributions found that, among all the tested models, the combination of the Becke-Perdew functional and a polarized valence basis set of triple-zeta quality provides the best representation of the electron momentum distributions for all of the 20 valence orbitals of norbornane. This experimentally validated quantum chemistry model was then used to extract some chemically important properties of norbornane. When these calculated properties are compared to corresponding results from other independent measurements, generally good agreement is found. Green's function calculations with the aid of the third-order algebraic diagrammatic construction scheme indicate that the orbital picture of ionization breaks down at binding energies larger than 22.5 eV. Despite this complication, they enable insights within 0.2 eV accuracy into the available ultraviolet photoemission and newly presented (e,2e) ionization spectra, except for the band associated with the 1a(2) (-1) one-hole state, which is probably subject to rather significant vibronic coupling effects, and a band at approximately 25 eV characterized by a momentum distribution of "s-type" symmetry, which Green's function calculations fail to reproduce. We note the vicinity of the vertical double ionization threshold at approximately 26 eV. PMID- 15549937 TI - A multichannel electron energy loss spectrometer for low-temperature condensed films. AB - We describe a wide-gap multichannel cylindrical deflection electron energy analyzer suitable for measuring the weak signals characteristic of electronically inelastic electron energy loss spectra. The analyzer has nearly ideal fringing field termination, and its resolution and energy dispersion were characterized as a function of energy by solving numerically the equation of motion of electrons in an ideal cylindrical electric field. The numerical results for the radial location of the electrons at the detector as a function of the entrance location, angle, and energy are closely approximated by a second order polynomial, and match closely with those observed. The detection efficiency of the analyzer is 100-150 times better than that of an equivalent single-channel instrument, but limited energy transmission of the zoom lens system used in our case reduced it by a factor of about 2. The performance of the new instrument was demonstrated by measuring the (3)E(1u) electronic spectrum of benzene in only 2 min and the spectrum of endo-benzotricyclo[4.2.1.0(2.5)]nonane. PMID- 15549938 TI - Joint experimental and theoretical study of vibrationally inelastic electron scattering on propane. AB - Vibrational electron energy loss spectra were measured for propane at incident energies of 3, 6, 10, 15, 20, and 25 eV at scattering angles of 40 degrees, 55 degrees, 70 degrees, 85 degrees, and 100 degrees . The spectra are compared with the results of ab initio calculations using a recently developed two-channel discrete momentum representation method. Good agreement between theory and experiment was found for large scattering angles and energies above the resonant region. PMID- 15549939 TI - Probing electronic coupling in excitonically coupled heterodimer complexes by two color three-pulse photon echoes. AB - Following the earlier work of Yang et al. [J. Chem. Phys. 110 (1999) 2983] analytical expressions for the downhill and uphill resonant two-color three-pulse photon echo peak shift (2C-3PEPS) of a heterodimer system are derived in the impulsive limit. It is shown how to obtain information about coupling between the components of the dimer from the combined one- and two-color peak shift measurements. Further analytical relations are derived which enable site specific information about the environment of the components, including the relative difference of the inhomogeneity and the difference between the energy-gap correlation functions on the heterodimer sites to be obtained. The simulations show only a very small influence of the laser pulse length on the measured values of coupling coefficient and other relevant quantities suggesting that current 2C 3PEPS measurements can find practical application in directly measuring couplings in excitonically coupled heterodimer complexes. PMID- 15549940 TI - Dynamical and structural properties of benzene in supercritical water. AB - We have employed an anisotropic united atom model of benzene (R. O. Contreras, Ph.D. thesis, Universitat Rovira i Virgili 2002) that reproduces the quadrupolar moment of this molecule through the inclusion of seven point charges. We show that this kind of interaction is required to reproduce the solvation of these molecules in supercritical water. We have computed self-diffusion coefficient and Maxwell-Stefan coefficients as well as the shear viscosity for the mixture water benzene at supercritical conditions. A strong density and composition dependence of these properties is observed. In addition, our simulations are in qualitative agreement with the experimental evidence that, at medium densities (0.6 g/cm(3) and 673 K), almost half of the benzene molecules have one hydrogen bond with water molecules. We also observe that these bonds are longer lived than the corresponding hydrogen bonds between water molecules. Similarly, we obtain an important reduction of the dielectric constant of the mixture with the increment of the amount of benzene molecules at medium and high densities. PMID- 15549941 TI - Stochastic Liouville equation simulation of multidimensional vibrational line shapes of trialanine. AB - The line shapes detected in coherent femtosecond vibrational spectroscopies contain direct signatures of peptide conformational fluctuations through their effect on vibrational frequencies and intermode couplings. These effects are simulated in trialanine using a Green's function solution of a stochastic Liouville equation constructed for four collective bath coordinates (two Ramachandran angles affecting the mode couplings and two diagonal energies). We find that fluctuations of the Ramachandran angles which hardly affect the linear absorption can be effectively probed by two-dimensional spectra. The signal generated at k(1)+k(2)-k(3) is particularly sensitive to such fluctuations. PMID- 15549942 TI - Orientational dynamics of hydrogen-bonded phenol. AB - We use femtosecond mid-infrared pump-probe spectroscopy to study the effects of hydrogen bonding on the orientational dynamics of the OD-stretch vibration of phenol-d. We study two samples: phenol-d in chloroform and phenol-d in chloroform to which we added excess acetone. For phenol-d in chloroform, we observe rotational diffusion of the OD group around the CO bond, with a correlation time of 3.7 ps. For phenol-d hydrogen bonded to acetone, the reorientation time is strongly dependent on the probe frequency, varying from 3 ps on the blue side of the spectrum to more than 30 ps on the red side. (c) 2004 American Institute of Physics. PMID- 15549943 TI - First-order mean spherical approximation for inhomogeneous fluids. AB - The first-order mean-spherical approximation (FMSA) [Y. Tang, J. Chem. Phys., 118, 4140 (2003)] is extended to the studies of inhomogeneous fluids by combining with Rosenfeld's perturbative method [Y. Rosenfeld, J. Chem. Phys. 98, 8126 (1993)]. In the extension, the key input-direct correlation function of FMSA-is applied to constructing the free energy density functional. Preserving its high fidelity at the bulk limit, the FMSA shows satisfactory performance for Yukawa fluids near hard and attractive walls. The results are better than or comparable to several other theories reported before for the geometry. The FMSA is found, in particular, more satisfactory than the traditional mean-field theory for predicting density profiles around hard walls. The FMSA is also compared with the full MSA for inhomogeneous fluids, showing no appreciable differences. The inhomogeneous FMSA goes successfully through the self-consistency test for reproducing the radial distribution function of the bulk Yukawa fluid. As far as the computation is concerned, the FMSA can be executed much faster than any nonmean-field theories, and the speed is virtually identical to that of the mean field theory. PMID- 15549944 TI - Surface hopping simulation of the vibrational relaxation of I2 in liquid xenon using the collective probabilities algorithm. AB - A surface hopping simulation of the vibrational relaxation of highly excited I(2) in liquid xenon is presented. The simulation is performed by using the collective probabilities algorithm which assures the coincidence of the classical and quantum populations. The agreement between the surface hopping simulation results and the experimental measurements for the vibrational energy decay curves at different solvent densities and temperatures is shown to be good. The overlap of the decay curves when the time axis is linearly scaled is explained in terms of the perturbative theory for the rate constants. The contribution of each solvent atom to the change of the quantum populations of the solute molecule is used to analyze the mechanism of the relaxation process PMID- 15549945 TI - Hydrogen storage in LiAlH4: predictions of the crystal structures and reaction mechanisms of intermediate phases from quantum mechanics. AB - We use the density functional theory and x-ray and neutron diffraction to investigate the crystal structures and reaction mechanisms of intermediate phases likely to be involved in decomposition of the potential hydrogen storage material LiAlH(4). First, we explore the decomposition mechanism of monoclinic LiAlH(4) into monoclinic Li(3)AlH(6) plus face-centered cubic (fcc) Al and hydrogen. We find that this reaction proceeds through a five-step mechanism with an overall activation barrier of 36.9 kcal/mol. The simulated x ray and neutron diffraction patterns from LiAlH(4) and Li(3)AlH(6) agree well with experimental data. On the other hand, the alternative decomposition of LiAlH(4) into LiAlH(2) plus H(2) is predicted to be unstable with respect to that through Li(3)AlH(6). Next, we investigate thermal decomposition of Li(3)AlH(6) into fcc LiH plus Al and hydrogen, occurring through a four-step mechanism with an activation barrier of 17.4 kcal/mol for the rate-limiting step. In the first and second steps, two Li atoms accept two H atoms from AlH(6) to form the stable Li-H-Li-H complex. Then, two sequential H(2) desorption steps are followed, which eventually result in fcc LiH plus fcc Al and hydrogen: Li(3)AlH(6)(monoclinic)-->3 LiH(fcc)+Al(fcc)+3/2 H(2) is endothermic by 15.8 kcal/mol. The dissociation energy of 15.8 kcal/mol per formula unit compares to experimental enthalpies in the range of 9.8-23.9 kcal/mol. Finally, we explore thermal decomposition of LiH, LiH(s)+Al(s)- >LiAl(s)+12H(2)(g) is endothermic by 4.6 kcal/mol. The B32 phase, which we predict as the lowest energy structure for LiAl, shows covalent bond characters in the Al-Al direction. Additionally, we determine that transformation of LiH plus Al into LiAlH is unstable with respect to transformation of LiH through LiAl. PMID- 15549946 TI - Evaluation of small-angle x-ray scattering data of a Raney-type Ni catalyst with computer simulation. AB - A reverse Monte Carlo-type simulation method was developed for the evaluation of anomalous small-angle x-ray scattering data of a Raney-type Ni catalyst. Based on other experimental data the catalytic Ni particles were modeled as small crystalline cylinders dispersed in the matrix. The average size of the Ni particles and their pair-correlation function were determined. Despite the unknown density of the catalyst, it is shown that each particle has about 2 neighbors in the first neighboring shell independent of the modeling density, and the position of the first peak of the pair-correlation function does not depend on the modeling density. A method was elaborated to get reasonable performance of the Reverse Monte Carlo-type simulation. The scattered intensity was calculated on the basis of probe scattering atoms put inside the cylinders. The effects of the omission of the real number of the atoms, the unknown density, the lack of normalization and the uncertainties in the cross sections were unified in two constants that were determined during the simulation. The method can be used for nanoparticles with other shape, where analytic form factors are complicated, and it may be powerful in the investigation of the usually neglected or simplified inter-particle structure of these systems. PMID- 15549947 TI - Nanospheres in phase-separating multicomponent fluids: a three-dimensional dissipative particle dynamics simulation. AB - The dynamics of phase separation of three-dimensional fluids containing nanospheres, which interact preferentially with one of the two fluids, is studied by means of large-scale dissipative particle dynamics simulations. We systematically investigated the effect of volume fraction, radius, and mass of the nanoparticles on both kinetics and morphology of the binary mixture. We found that nanospheres lead to a reduction of domain growth which is intensified as their volume fraction is increased for a given radius of nanoparticles, or as the nanoparticles radius is decreased for a given volume fraction. Up to moderate volume fractions of nanoparticles, the growth law, however, is found to be identical to that pure binary fluids, i.e., R(t) approximately t(n), with n=1. For relatively high volume fractions of nanoparticles, a diffusive growth regime was detected. The crossover to the slower growth regime as the nanoparticles volume fraction is increased or their radius is decreased is associated with the crystallization of the nanospheres within the preferred component. These results are qualitatively in good agreement with previous two-dimensional simulations using molecular dynamics [M. Laradji and G. MacNevin, J. Chem. Phys. 119, 2275 (2003)] and a time-dependent Ginzburg-Landau model [M. Laradji, J. Chem. Phys. 120, 9330 (2004)], as well as recent experiments. PMID- 15549948 TI - Molecular exchange dynamics in partially filled microscale and nanoscale pores of silica glasses studied by field-cycling nuclear magnetic resonance relaxometry. AB - Nuclear magnetic spin-lattice relaxation experiments have been performed in partially filled porous glasses with wetting and nonwetting fluids. The frequency dependence of the spin-lattice relaxation rate in Vycor (4 nm pores) and VitraPOR #5 (1 microm pores) silica glasses was studied as a function of the filling degree with the aid of field-cycling NMR relaxometry. The species of primary interest were water ("polar") and cyclohexane ("nonpolar"). Spin-lattice relaxation was examined in the frequency range from 1 kHz to 400 MHz with the aid of a field-cycling NMR relaxometer and an ordinary 400 MHz NMR spectrometer. Three different mobility states of the fluid molecules are distinguished: The adsorbed state at the pore walls, the bulklike liquid phase, and the vapor phase. The adsorbate spin-lattice relaxation rate is dominated by the "reorientation mediated by translational displacements" (RMTD) mechanism taking place at the adsorbate/matrix interface at frequencies low enough to neglect rotational diffusion of the molecules. The experimental data are analyzed in terms of molecular exchange between the different mobility states. Judged from the dependence of the spin-lattice relaxation rates on the filling degree, limits for slow and fast exchange (relative to the RMTD time scale) can be distinguished and identified. It is concluded that water always shows the features of slow exchange irrespective of the investigated pore sizes and filling degrees. This is in contrast to cyclohexane which is subject to slow exchange in micrometer pores, whereas fast exchange occurs in nanoscopic pores. The latter case implies that the vapor phase contributes to molecular dynamics in this case at low filling degrees while it is negligible otherwise. PMID- 15549949 TI - Surface-enhanced Raman scattering from ordered Ag nanocluster arrays. AB - We have examined the effect of ordered silver nanocluster substrates on the surface-enhanced Raman spectrum of rhodamine 6G (R6G). Triangular shaped silver nanocluster arrays with order on the approximately 100 mum range were prepared using nanosphere lithography. Direct comparisons of R6G surface-enhanced Raman spectroscopy (SERS) signals between ordered nanocluster regions and amorphous Ag regions prepared under identical deposition conditions provide strong evidence of an electromagnetic field enhancement attributed to the unique nanocluster morphology. We have obtained order of magnitude enhancement factors for both 200 and 90 nm Ag nanocluster SERS substrates relative to Ag films. PMID- 15549950 TI - HREELS, STM, and STS study of CH(3)-terminated Si(111)-(1x1) surface. AB - An ideally (1x1)-CH(3)(methyl)-terminated Si(111) surface was composed by Grignard reaction of photochlorinated Si(111) and the surface structure was for the first time confirmed by Auger electron spectroscopy, low energy electron diffraction, high-resolution electron energy loss spectroscopy (HREELS), scanning tunneling microscopy (STM), and scanning tunneling spectroscopy (STS). HREELS revealed the vibration modes associated to the CH(3)-group as well as the C-Si bond. STM discerned an adlattice with (1x1) periodicity on Si(111) composed of protrusions with internal features, covering all surface terraces. The surface structure was confirmed to be stable at temperatures below 600 K. STS showed that an occupied-state band exists at gap voltage of -1.57 eV, generated by the surface CH(3) adlattice. This CH(3):Si(111)-(1x1) adlayer with high stability and unique electronic property is prospective for applications such as nanoscale lithography and advanced electrochemistry. PMID- 15549951 TI - Calculating the hopping times of confined fluids: two hard disks in a box. AB - The dynamical transition between the anomalous single file diffusion of highly confined fluids and bulk normal diffusion can be described by a phenomenological model involving a particle hopping time tau(hop). We suggest a theoretical formalism that will be useful for the calculation of tau(hop) for a variety of systems and test it using a simple model consisting of two hard disks confined to a rectangular box with hard walls. In the case where the particles are moving diffusively, we find the hopping time diverges as a power law in the threshold region with an exponent of -(3/2). Under conditions where the particles move inertially, transition state theory predicts a power law behavior with an exponent of -2. Molecular dynamics simulations confirm the transition state theory result for inertial dynamics, while Brownian dynamics simulations suggest the scaling exponent is highly sensitive to the details of the algorithm. PMID- 15549952 TI - Calculation of pressure using the virtual-volume-variation method and the virial method from chain conformations obtained by Monte Carlo simulations on the second nearest neighbor diamond lattice. AB - For a model system of polyethylene of chain lengths 40 and 100 carbon atoms, we calculated the pressure at different densities and compared them with the experimental values. The simulation was conducted on the second nearest neighbor diamond lattice, and the pressure was calculated using the virtual-volume variation method after the system was reverse mapped to its fully atomistic form in continuous space and energy minimized. In addition, the pressure was also calculated from the virial route by conducting a short molecular dynamics simulation starting from the energy minimized structure. We show that the pressure obtained from our simulations is quite reasonable in the length of simulation time (in Monte Carlo steps) normally employed in our group. These results provide additional evidence for the equilibration of our model systems, and methodology to calculate the pressure in our lattice models. PMID- 15549953 TI - Nanotubular boron-carbon heterojunctions. AB - Linear nanotubular boron-carbon heterojunctions are systematically constructed and studied with the help of ab initio total energy calculations. The structural compatibility of the two classes of materials is shown, and a simple recipe that determines all types of stable linear junctions is illustrated in some detail. Our results also suggest the compatibility of various technologically interesting types of nanotubular materials, leading to different types of nanotubular compound materials, and pointing out the possibility of wiring nanotubular devices within heterogeneous nanotubular networks. PMID- 15549954 TI - Optical spectra and localization of excitons in inhomogeneous helical cylindrical aggregates. AB - We study the linear optical properties of helical cylindrical molecular aggregates accounting for the effects of static diagonal disorder. Absorption, linear dichroism, and circular dichroism spectra are presented, calculated using brute force numerical simulations and a modified version of the coherent potential approximation that accounts for finite size effects by using the appropriate open boundary conditions. Excellent agreement between both approaches is found. It is also shown that the inclusion of disorder results in a better agreement between calculated and measured spectra for the chlorosomes of green bacteria as compared to our previous report, where we restricted ourselves to homogeneous cylinders [Didraga, Klugkist, and Knoester, J. Phys. Chem. B 106, 11474 (2002)]. For the excitons that govern the optical response, we also investigate the disorder-induced localization properties. By analyzing an autocorrelation function of the exciton wave function, we find a strongly anisotropic localization behavior, closely following the properties of chiral wave functions which previously have been found for homogenoeus helical cylinders [Didraga and Knoester, J. Chem. Phys. 121, 946 (2004)]. It is shown that the circular dichroism spectrum may still show a strong dependence on the cylinder length, even when the exciton wave function is localized in a region small compared to the cylinder's size. PMID- 15549955 TI - Long-time dynamics of Rouse-Zimm polymers in dilute solutions with hydrodynamic memory. AB - The dynamics of flexible polymers in dilute solutions is studied taking into account the hydrodynamic memory, as a consequence of fluid inertia. As distinct from the Rouse-Zimm (RZ) theory, the Boussinesq friction force acts on the monomers (beads) instead of the Stokes force, and the motion of the solvent is governed by the nonstationary Navier-Stokes equations. The obtained generalized RZ equation is solved approximately using the preaveraging of the Oseen tensor. It is shown that the time correlation functions describing the polymer motion essentially differ from those in the RZ model. The mean-square displacement (MSD) of the polymer coil is at short times approximately t(2) (instead of approximately t). At long times the MSD contains additional (to the Einstein term) contributions, the leading of which is approximately t. The relaxation of the internal normal modes of the polymer differs from the traditional exponential decay. It is displayed in the long-time tails of their correlation functions, the longest lived being approximately t(-3/2) in the Rouse limit and t(-5/2) in the Zimm case, when the hydrodynamic interaction is strong. It is discussed that the found peculiarities, in particular, an effectively slower diffusion of the polymer coil, should be observable in dynamic scattering experiments. PMID- 15549956 TI - Excitation energies of pi-conjugated oligomers within time-dependent current density-functional theory. AB - We study the pi*<--pi singlet excitations of the pi-conjugated oligomers of polyacetylene, polydiacetylene, polybutatriene, polythiophene, poly(para phenylene vinylene), and the lowest singlet excitations of the hydrogen chain. For this we used time-dependent current-density-functional theory within the Vignale-Kohn and adiabatic local density approximations. By studying the dependence of the excitation spectrum on the chain length we conclude that the reduction of the static polarizability when using the Vignale-Kohn functional has two origins. First, the excitation energies of transitions with a large transition dipole are shifted upward. Second, the character of the transition between the lowest occupied and highest unoccupied molecular orbitals and the oscillator strength of the lowest transition within the adiabatic local density approximation is transferred to higher transitions. The lowest transitions that have a considerable oscillator strength obtained with the Vignale-Kohn functional have excitation energies that are in most cases in better agreement with available reference data than the adiabatic local density approximation. PMID- 15549957 TI - Thermodynamic properties of Lennard-Jones chain molecules: renormalization-group corrections to a modified statistical associating fluid theory. AB - A modified version of the statistical associating fluid theory (SAFT), the so called soft-SAFT equation of state (EOS), has been extended by a crossover treatment to take into account the long density fluctuations encountered when the critical region is approached. The procedure, based on White's work from the renormalization group theory [Fluid Phase Equilibria 75, 53 (1992); L. W. Salvino and J. A. White, J. Chem. Phys. 96, 4559 (1992)], is implemented in terms of recursion relations where the density fluctuations are successively incorporated. The crossover soft-SAFT equation provides the correct nonclassical critical exponents when approaching the critical point, and reduces to the original soft SAFT equation far from the critical region. The accuracy of the global equation is tested by direct comparison with molecular simulation results of Lennard-Jones chains, obtaining very good agreement and clear improvements compared to the original soft-SAFT EOS. Excellent agreement with vapor-liquid equilibrium experimental data inside and outside the critical region for the n-alkane series is also obtained. We provide a set of transferable molecular parameters for this family, unique for the whole range of thermodynamic properties. PMID- 15549958 TI - Designing generalized statistical ensembles for numerical simulations of biopolymers. AB - Conformational properties of polymers, such as average dihedral angles or molecular alpha-helicity, display a rather weak dependence on the detailed arrangement of the elementary constituents (atoms). We propose a computer simulation method to explore the polymer phase space using a variant of the standard multicanonical method, in which the density of states associated to suitably chosen configurational variables is considered in place of the standard energy density of states. This configurational density of states is used in the Metropolis acceptance/rejection test when configurations are generated with the help of a hybrid Monte Carlo algorithm. The resulting configurational probability distribution is then modulated by exponential factors derived from the general principle of the maximal constrained entropy by requiring that certain average configurational quantities take preassigned (possibly temperature dependent) values. Thermal averages of other configurational quantities can be computed by using the probability distributions obtained in this way. Moments of the energy distribution require an extra canonical sampling of the system phase space at the desired temperature, in order to locally thermalize the configurational degrees of freedom. As an application of these ideas we present the study of the structural properties of two simple models: a bead-and-spring model of polyethylene with independent hindered torsions and an all-atom model of alanine and glycine oligomers with 12 amino acids in vacuum. PMID- 15549959 TI - Phase evolution of lamellar cationic lipid-DNA complex: steric effect of an electrolyte. AB - The complexation isotherms of DNA plus lipids of a symmetric lamellar cationic lipid-DNA system were determined within a mean field free energy. The free energy incorporates the ion's finite size of NaCl simple electrolyte in solution and makes use of known structure data on this complex. The results for the predicted isotherms are in qualitative agreement with the trends of the experimental data for this property. PMID- 15549960 TI - Replica exchange molecular dynamics simulations of amyloid peptide aggregation. AB - The replica exchange molecular dynamics (REMD) approach is applied to four oligomeric peptide systems. At physiologically relevant temperature values REMD samples conformation space and aggregation transitions more efficiently than constant temperature molecular dynamics (CTMD). During the aggregation process the energetic and structural properties are essentially the same in REMD and CTMD. A condensation stage toward disordered aggregates precedes the beta-sheet formation. Two order parameters, borrowed from anisotropic fluid analysis, are used to monitor the aggregation process. The order parameters do not depend on the peptide sequence and length and therefore allow to compare the amyloidogenic propensity of different peptides PMID- 15549961 TI - Conformation of a polymer chain in solution: an exact density expansion approach. AB - The conformation of a polymer chain in solution is intrinsically coupled to the thermodynamic and structural properties of the solvent. Here we study such solvent effects in a system consisting of a flexible interaction-site n-mer chain immersed in a monomeric solvent. Chain conformation is described with a set of intramolecular site-site probability functions. We derive an exact density expansion for these intramolecular probability functions and give a diagrammatic representation of the terms contributing at each order of the expansion. The expansion is tested for a short hard-sphere chain (n=3 or 4) with site diameter sigma in a hard-sphere solvent with solvent diameter D. In comparison with Monte Carlo simulation results for 0.2< or =D/sigma< or =100, the expansion (taken to second order) is found to be quantitatively accurate for low to moderate solvent volume fractions for all size ratios. Average chain dimensions are predicted accurately up to liquidlike solvent densities. The hard-sphere chains are compressed with both increasing solvent density and decreasing solvent size. For small solvent (D100 nm) diffusion significantly disrupted by polymer chains showing an approximately 30% reduction in diffusion coefficient. The transition from the short diffusion to the long one occurs in a very narrow range (10-100 nm) of diffusion distance and this unique character of HA realizing anomalous diffusion should provide suitable environments for various bioactivities when involved in ECM. PMID- 15549965 TI - Self-diffusion in solutions of a 20 base pair oligonucleotide: effects of concentration and ionic strength. AB - The long-time self-diffusion coefficients of a 20 base pair duplex oligonucleotide are measured as functions of 20-mer and added NaCl salt concentrations. The self-diffusion coefficients decrease monotonically with increasing 20-mer concentrations for the high-added salt sample and display non monotonically decreasing 20-mer concentration dependences at lower added salt concentrations. The non-monotonic behavior is attributed to the opposing effects of the tendency to increase the interactions between 20-mers as the concentration is increased and to a decrease in the extent of the Coulomb forces as counterions from the 20-mer increasingly screen them. Attempts to account for the effect of the Coulomb forces on the self-diffusion coefficients by using effective dimensions in the hard rod theory give good agreement with experiment at the highest salt concentration studied. For the lower salt concentrations there appear to be two scaling regimes--one at low polyion concentration in which the high salt scaling of the rod dimensions by adding the Debye screening to the length and diameter of the rod is appropriate and one at high polyion concentrations where the scaling of the dimensions is the addition of 1/2 the Debye screening length. Estimates of the "overlap" concentration C*=1/L(eff) indicate that the non-monotonic decrease occurs at concentrations lower than C*. Finally, the fluorescence correlation spectroscopy self-diffusion coefficients measured here are compared with the mutual diffusion coefficients measured by dynamic light scattering. PMID- 15549966 TI - Triplet build in and decay of isolated polyspirobifluorene chains in dilute solution. AB - The triplet kinetics of a conjugated polymer, polyspirobifluorene, have been studied using time resolved photoinduced absorption spectroscopy and gated emission delayed fluorescence. Working on isolated polymer chains in dilute solution, we pay particular attention to the buildup and decay of the triplet states following intersystem crossing from the excited singlet state. Confirmation of intersystem crossing as a monomolecular cold process has been made. At high excitation powers an initial fast decay of the triplet has been observed; this is attributed to intrachain triplet-triplet annihilation. From this observation we estimate the lower bound of the intersystem crossing yield as 1.2%. We also calculate the intrachain annihilation constant to be (2.9+/-0.1)x 10(8) cm(3) s(-1). PMID- 15549967 TI - Concentration dependent size of reversibly assembling polymers in solution: a mean-field lattice theory. AB - The concentration dependence of the mean length of equilibrium polymers, , as a function of solute volume fraction is studied in a lattice description. Using a more detailed model of constituent interactions in comparison to previous studies we are able to find conditions under which a decrease of at high solute concentration may occur. PMID- 15549968 TI - Monte Carlo simulation of structure and nanoscale interactions in polymer nanocomposites. AB - Off-lattice Monte Carlo simulations in the canonical ensemble are used to study polymer-particle interactions in nanocomposite materials. Specifically, nanoscale interactions between long polymer chains (N=550) and strongly adsorbing colloidal particles of comparable size to the polymer coils are quantified and their influence on nanocomposite structure and dynamics investigated. In this work, polymer-particle interactions are computed from the integrated force-distance curve on a pair of particles approaching each other in an isotropic polymer medium. Two distinct contributions to the polymer-particle interaction potential are identified: a damped oscillatory component that is due to chain density fluctuations and a steric repulsive component that arises from polymer confinement between the surfaces of approaching particles. Significantly, in systems where particles are in a dense polymer melt, the latter effect is found to be much stronger than the attractive polymer bridging effect. The polymer particle interaction potential and the van der Waals potential between particles determine the equilibrium particle structure. Under thermodynamic equilibrium, particle aggregation is observed and there exists a fully developed polymer particle network at a particle volume fraction of 11.3%. Near-surface polymer chain configurations deduced from our simulations are in good agreement with results from previous simulation studies. PMID- 15549969 TI - Intestinal candyfloss. AB - In this issue of the Biochemical Journal, Robbe et al. have employed a state-of the-art MS technique to re-examine the sweet and sticky sugar covering of human intestinal mucosa. The availability of highly sensitive MS methods has been instrumental in determining the complex structures of mucin oligosaccharides, and in demonstrating that there are variations in structure along the sections of the gut. In contrast to previous studies, these results described by Robbe et al. show some correlation with the activities of enzymes synthesizing mucin oligosaccharides in the human colon. Many questions still remain to be answered: for example, regarding the regulation of the relative amounts of sugar chains in individual mucins, and their roles in the homoeostasis of the intestinal mucosa. Structural studies are a solid basis for understanding the functions of sugar chains, and the mechanisms and significance of changes during the development of intestinal disease. PMID- 15549970 TI - The Nazi hypothermia experiments: forbidden data? PMID- 15549971 TI - Early thoughts on 'Why Mothers Die 2000-2002'. PMID- 15549972 TI - The prognostic value of pre-operative predicted forced vital capacity in corrective spinal surgery for Duchenne's muscular dystrophy. AB - The majority of patients with Duchenne's muscular dystrophy require corrective spinal surgery for scoliosis to maintain seated balance and to slow the progression of respiratory compromise, thereby facilitating nursing and enhancing their quality of life. Traditionally patients with a pre-operative forced vital capacity (PFVC) of 30% or below predicted have been denied this surgery as it was thought that the incidence of postoperative complications was unacceptably high. We present data collected prospectively from 45 consecutive operations undertaken in our unit. These cases indicate that there is no clinically significant difference in operative and postoperative outcomes between patients with PFVC > 30% and < or =30%. However, the routine postoperative use of mask ventilation to facilitate early tracheal extubation is vital. PMID- 15549973 TI - Ease of insertion of the laryngeal tube during manual-in-line neck stabilisation. AB - The laryngeal tube has a potential role in airway management during anaesthesia or cardiopulmonary resuscitation. In patients with unstable necks, the head and neck may need to be stabilised manually (manual in-line stabilisation), but it is not known whether this procedure affects the ease of insertion of the laryngeal tube. We studied, in a cross-over study, 21 adult patients to compare the success rate of ventilation through the laryngeal tube between the Magill position (a pillow under the occiput and the head extended) or the manual in-line position of the head and neck (without a pillow under the occiput). After induction of anaesthesia and neuromuscular blockade, the laryngeal tube was inserted in turn in the two positions. The ease of insertion was scored with four categories (easy, moderately difficult, difficult and impossible), and adequacy of ventilation through the device was assessed. Ventilation was adequate in all 21 patients in the Magill position, but only in two of 21 patients during manual in line positioning (p < 0.01; 95%CI for difference: 68-94%). In the Magill position, insertion of the laryngeal tube was easy in 16 patients and moderately difficult in the remaining five patients; in the manual in-line stabilisation position, insertion was moderately difficult in two patients and impossible in the remaining 19 patients. Stabilisation of the patient's head and neck by the manual in-line method made insertion of the laryngeal tube either difficult or impossible. PMID- 15549974 TI - Comparison of Propofol-Lipuro with propofol mixed with lidocaine 10 mg on propofol injection pain. AB - A common drawback of propofol is pain on injection and lidocaine is commonly mixed with propofol to reduce its incidence and severity. We conducted a randomised, prospective, double-blind study to compare injection pain following the administration of two different formulations of propofol in 200 unpremedicated ASA I-III adult patients scheduled for elective surgery under general anaesthesia. Patients were allocated randomly into two groups to receive either Propofol-Lipuro without added lidocaine or Diprivan mixed with lidocaine 10 mg. Five ml of the study solution was injected at a constant rate over 15 s and patients graded any associated pain or discomfort using a four-point verbal rating scale. The incidence of propofol injection pain was virtually identical in both study groups with 37/98 (38%) patients experiencing pain or discomfort following Propofol-Lipuro compared with 35/98 (36%) after Diprivan (p = 0.88). We observed no significant difference in pain scores between the groups (p = 0.67). Moderate or severe injection pain was experienced by 12/98 (12%) patients given Propofol-Lipuro compared with 8/98 (8%) given Diprivan (p = 0.48). PMID- 15549975 TI - Cardiovascular response to acute normovolaemic haemodilution in patients with severe aortic stenosis: assessment with transoesophageal echocardiography. AB - Using multiplane transoesophageal echocardiography (TOE), we investigated the haemodynamic response to acute normovolaemic haemodilution (ANH) in anaesthetised patients with critical aortic stenosis. Twenty-eight patients were randomly assigned to ANH or control groups. In the control group, haemodynamic data remained unchanged over a 20-min period. In the ANH group, haemoglobin levels decreased from a mean (SD) of 134 (7) to 91 (9) g x l(-1) (p < 0.001) whereas stroke volume, central venous pressure and left ventricular (LV) end-diastolic area all increased significantly (mean (SD) +15 (6) ml; +2.0 (1.1) mmHg; +2.1 (0.8) cm2, respectively). During ANH, the accelerated blood flow through the stenotic valve caused an increased loss (SD) in LV stroke work: from 24 (8)% to 30 (10)%), (p < 0.01). Hence, lowering viscosity with ANH resulted in improved venous return, higher cardiac preload and increased stroke volume. However, this adaptive haemodynamic response was limited by less efficient LV stroke work due to dissipation of fluid kinetic energy. PMID- 15549976 TI - Cardiovascular effects of xenon and nitrous oxide in patients during fentanyl midazolam anaesthesia. AB - Xenon anaesthesia appears to have minimal haemodynamic effects. The purpose of this randomised prospective study was to compare the cardiovascular effects of xenon and nitrous oxide in patients with known ischaemic heart disease. In 20 patients who were due to undergo coronary artery bypass graft surgery, 30 min following induction of anaesthesia with fentanyl 30 microg x kg(-1) and midazolam 0.1 mg x kg(-1) but prior to the start of surgery, xenon or nitrous oxide 60% was administered for 15 min. The results showed that xenon caused a minimal decrease in the mean arterial pressure (from 81 (7) to 75 (8) mmHg, mean (SD)), but did not affect the systolic function of the left ventricle, as demonstrated by unchanged left ventricular stroke work index (LVSWI) and the fractional area change of the left ventricle (FAC) derived from transoesophageal echocardiography (TOE). However, in contrast, nitrous oxide was found to decrease the mean arterial pressure (from 81 (8) to 69 (7) mmHg), the LVSWI, and the FAC. The cardiac index, central venous and pulmonary artery occlusion pressures, systemic and pulmonary vascular resistances, and the TOE-derived E/A ratio through the mitral valve were unchanged by xenon or nitrous oxide. We conclude that xenon provides improved haemodynamic stability compared with nitrous oxide, conserving the left ventricular systolic function. PMID- 15549977 TI - Transoesophageal echocardiography is unreliable for cardiac output assessment after cardiac surgery compared with thermodilution. AB - This randomised, single-blind, double-control study compared and established prospectively the best transoesophageal echocardiography methods for determining cardiac output in patients after cardiac surgery. Thirty patients undergoing coronary artery bypass grafting were included. Measurements were taken postoperatively, after stabilisation in the intensive care unit. Cardiac output was determined by transoesophageal echocardiography in randomised order through the aortic, mitral, and pulmonary valves, right and left ventricular outflow tracts, transgastric surface areas of the left ventricle and left ventricle two dimensional volumes (Simpson's rules). 'Eyeball guessing' was done off-line. The best results were transaortic measurements using the triangular shape assumption of valve opening, but some values deviated considerably, and none of these approaches reached the limit of agreement set at 30% when compared to thermodilution. Eyeball guessing was comparable to the best transoesophageal echocardiography measurements. We conclude that transoesophageal echocardiography is an unreliable tool for determination of cardiac output in intensive care after cardiac surgery. PMID- 15549978 TI - Prescription errors in UK critical care units. AB - Drug prescription errors are a common cause of adverse incidents and may be largely preventable. The incidence of prescription errors in UK critical care units is unknown. The aim of this study was to collect data about prescription errors and so calculate the incidence and variation of errors nationally. Twenty four critical care units took part in the study for a 4-week period. The total numbers of new and re-written prescriptions were recorded daily. Errors were classified according to the nature of the error. Over the 4-week period, 21,589 new prescriptions (or 15.3 new prescriptions per patient) were written. Eighty five per cent (18,448 prescriptions) were error free, but 3141 (15%) prescriptions had one or more errors (2.2 erroneous prescriptions per patient, or 145.5 erroneous prescriptions per 1000 new prescriptions). The five most common incorrect prescriptions were for potassium chloride (10.2% errors), heparin (5.3%), magnesium sulphate (5.2%), paracetamol (3.2%) and propofol (3.1%). Most of the errors were minor or would have had no adverse effects but 618 (19.6%) errors were considered significant, serious or potentially life threatening. Four categories (not writing the order according to the British National Formulary recommendations, an ambiguous medication order, non-standard nomenclature and writing illegibly) accounted for 47.9% of all errors. Although prescription rates (and error rates) in critical care appear higher than elsewhere in hospital, the number of potentially serious errors is similar to other areas of high-risk practice. PMID- 15549979 TI - Pharmacokinetics of sevoflurane uptake into the brain. AB - Two recent studies have examined the pharmacokinetics of sevoflurane in adults. Lu et al.(Pharmacokinetics of sevoflurane uptake into the brain and body, Anaesthesia 2003; 58: 951-6) observed that jugular bulb sevoflurane concentration initially rose unexpectedly rapidly and then approached arterial concentrations unexpectedly slowly, suggesting that a blood-brain diffusion barrier exists. They also observed a large alveolar-arterial sevoflurane gradient, suggesting that an alveolar-arterial diffusion barrier exists. Nakamura et al. (Predicted sevoflurane partial pressure in the brain with an uptake and distribution model: Comparison with the measured value in internal jugular vein blood. Journal of Clinical Monitoring and Computing 1999; 15: 299-305) found no diffusion barriers. We used a computer model to analyse both data sets and show that the observations of Lu et al. can be explained by contamination of jugular samples with extracerebral blood. It is possible that the alveolar-arterial gradients observed by Lu et al. are due to discrepancies in conversions between blood concentrations and gas partial pressures. Our study suggests that there is no blood-brain diffusion barrier for sevoflurane and that the data of Lu et al. must be interpreted with caution. PMID- 15549980 TI - Tracheal intubation using the Bonfils intubation fibrescope after failed direct laryngoscopy. AB - Failed tracheal intubation due to a difficult airway is an important cause of anaesthetic morbidity and mortality. This study was undertaken to evaluate the effectiveness of the Bonfils intubation fibrescope for tracheal intubation after failed direct laryngoscopy. Twenty-five patients undergoing coronary artery bypass grafting were enrolled in the study after two attempts at conventional laryngoscopy by a board certified anaesthetist had failed. Intubation with the Bonfils fibrescope was successful on the first attempt in 22 patients (88%) and on the first or second attempt in 24 patients (96%); in one patient intubation was impossible. Median (IQR [range]) time to intubation using the Bonfils intubation fibrescope was 47.5 (30-80 [20-200]) s. Tracheal intubation using the Bonfils intubation fibrescope appears to be a simple and effective technique for the management of a difficult intubation. PMID- 15549981 TI - Use of alpha-agonists for management of anaphylaxis occurring under anaesthesia: case studies and review. AB - Anaphylaxis is an uncommon but serious complication of anaesthesia. Most current guidelines for the management of anaphylaxis list only epinephrine as a vasopressor to use in the event of cardiovascular collapse. We present two cases of anaphylaxis under anaesthesia where return of spontaneous circulation was refractory to epinephrine, but occurred following the administration of the alpha agonist metaraminol. Potential advantages and disadvantages of using epinephrine in this setting, the role of alpha-agonists and some potential mechanisms accounting for their role in successful management are reviewed. PMID- 15549982 TI - Effective doses of vecuronium in a patient with myotonic dystrophy. AB - Of the forms of muscular dystrophy, myotonic dystrophy has the greatest systemic involvement. Although most patients with myotonic dystrophy show normal sensitivity to non-depolarising neuromuscular blocking drugs, some have been reported to show greatly increased sensitivity to these drugs, and little is known about the sensitivity of different muscles. We compared effective doses of vecuronium in a patient with myotonic dystrophy at the orbicularis oculi, adductor pollicis and flexor hallucis brevis muscles during total intravenous anaesthesia. The calculated ED50 for the orbicularis oculi (7.77 microg x kg(-1) (95% CI 3.10-16.8 microg x kg(-1))) was lower than for the adductor pollicis (25.3 microg x kg(-1) (95% CI 20.7-43.3 microg x kg(-1))) and flexor hallucis brevis muscles (29.5 microg x kg(-1) (95% CI 11.0-85.6 microg x kg(-1); p < 0.01)). The ED90 was also lower for the orbicularis oculi (35.7 microg x kg(-1) (95% CI 14.8-66.5 microg x kg(-1))) than for the other muscles (51.8 microg x kg( 1) (95% CI 29.3-145.0 microg x kg(-1)) and 50.6 microg x kg(-1) (95% CI 5.29 642.0 microg x kg(-1)), respectively) (p < 0.01)). PMID- 15549983 TI - Postoperative complications after CS spray exposure. AB - Summary We report on airway complications associated with general anaesthesia in a subject who had been exposed to CS spray several hours before surgery. CS spray is a form of tear gas that is said to have a short half-life when the subject is removed from exposure. Induction of anaesthesia was uneventful. Marked laryngospasm occurred when the tracheal tube was removed at the end of the operation, and the anaesthetists experienced lacrimation and burning sensations typical of CS exposure. The effects on the attending anaesthetist made tracheal re-intubation difficult. There were no long-term adverse sequelae for the patient or anaesthetists. Suggestions are made for changes to anaesthetic practice and the advice given by the police about patients who have been exposed to CS spray. PMID- 15549984 TI - Microbial keratitis in ITU staff: an occupational hazard? AB - Microbial keratitis is a potentially blinding corneal infection; the infection may progress rapidly if untreated and result in corneal perforation. The breakdown of the innate ocular defences is known to predispose to corneal infection. We present three cases of microbial keratitis in intensive therapy unit (ITU) staff each of whom had compromised corneal immunity. Anaesthetists and nurses regularly perform high-risk procedures, such as tracheal suctioning, which have been known to cause ocular infections by aerosol inoculation. We suggest that although the absolute risk of infection is low, susceptible individuals may be at increased risk of corneal infection from exposure to potentially pathogenic organisms in such environments. We stress the need for ITU staff, particularly those who wear contact lenses, to wear eye protection when performing procedures likely to cause infected aerosols. PMID- 15549985 TI - Anaesthetic and intensive care management of a patient with Ehlers-Danlos type IV syndrome after laparotomy. AB - A 31-year-old woman with Ehlers-Danlos type IV syndrome developed multiple intensive care related complications following laparotomy for perforated bowel. Complications are more likely to occur with the Ehlers-Danlos syndrome. PMID- 15549986 TI - Severe lactic acidosis following alcohol related generalised seizures. AB - A 45-year-old alcoholic man presented following several short grand-mal seizures. He was not known to be epileptic. Initial investigations demonstrated a severe lactic acidosis. The rise in lactate was one of the highest levels reported in similar patients. The patient recovered within 4 h of management with oxygen, fluids and sodium bicarbonate. Lactic acidosis following convulsions is often associated with spontaneous resolution and a favourable outcome. PMID- 15549987 TI - The history of spinal needles: getting to the point. AB - The history of the design of spinal needle tips is discussed, from the first needles used by J. Leonard Corning in 1885 to innovative, modern needle designs that continue to appear on the market. The shape of the needle tip started as a cutting bevel and developed into the atraumatic tip and the pencil-point tip in current common use. Innovative designs such as the stylet-tipped needle and the directional needle are described, as well as the needles used for continuous spinal anaesthesia. PMID- 15549988 TI - Difficult Airway Society guidelines. PMID- 15549989 TI - Difficult Airway Society guidelines. PMID- 15549990 TI - Difficult Airway Society guidelines. PMID- 15549991 TI - Difficult Airway Society guidelines. PMID- 15549992 TI - Difficult Airway Society guidelines. PMID- 15549993 TI - Professional conflicts in anaesthesia in the United States. PMID- 15549995 TI - Training and assessment of competency of trainees in the transfer of critically ill patients. PMID- 15549996 TI - Warming patients in the lithotomy position. PMID- 15549997 TI - Use of dornase alfa in the management of ARDS. PMID- 15549998 TI - Influence of the prone position on propofol pharmacokinetics. PMID- 15549999 TI - Reaction between parecoxib and vecuronium. PMID- 15550001 TI - Excess delivery of isoflurane liquid from a syringe driver. PMID- 15550002 TI - Sub-Tenon's block without hyaluronidase. PMID- 15550004 TI - Is euphoria a side-effect of lidocaine? PMID- 15550005 TI - Inadvertent overdose of muscle relaxants. PMID- 15550006 TI - Disruption of the oxygen supply to a heart-lung machine. PMID- 15550007 TI - Carbon dioxide in laryngeal mask airway cuffs. PMID- 15550009 TI - Use of a gum elastic bougie in the oesophagus to guide the ProSeal LMA. PMID- 15550011 TI - The early history and development of thrombolysis in acute myocardial infarction. PMID- 15550012 TI - Long-term warfarin use to prevent both stroke and dementia in subjects with atrial fibrillation? PMID- 15550013 TI - Dementia in subjects with atrial fibrillation: hemostatic function and the role of anticoagulation. AB - BACKGROUND: Atrial fibrillation (AF) is associated with cognitive impairment and dementia, perhaps through encouraging a prothrombotic state and cardioembolism. OBJECTIVES: We wished to test the hypotheses that hemostatic function is altered in subjects with AF who develop dementia, and that long-term warfarin anticoagulation is protective against this complication. PATIENTS AND METHODS: Recruitment was from an observational cohort study of AF. Baseline assessment included measurement of plasma fibrinogen, fibrin D-dimer, prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complexes (TAT), von Willebrand factor and tissue plasminogen activator. We assessed cognitive function after 3 years' follow-up using the 13-item modified Telephone Interview for Cognitive Status (TICSm) and the short form of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE). RESULTS: Of the 218 subjects assessed, 145 (66%) were prescribed warfarin. Forty-nine (22%) met TICSm/IQCODE criteria for dementia. D dimer, F1+2 and TAT levels were higher in AF subjects with dementia compared with those without (medians 81 vs. 60 ng mL(-1), P = 0.008; 0.76 vs. 0.49 nmol L(-1), P = 0.006; and 1.78 vs. 1.44 microg L(-1), P = 0.003, respectively). These associations became of borderline statistical significance following adjustment for age. Logistic regression showed a trend towards warfarin use being independently associated with reduced prevalence of dementia (odds ratio 0.52, P = 0.08). CONCLUSIONS: We found evidence of increased thrombin generation and fibrin turnover in subjects with AF and dementia compared with those without dementia. Long-term warfarin use may be protective against the development of dementia in subjects with AF. PMID- 15550014 TI - Towards a diagnosis of heparin-induced thrombocytopenia after cardiopulmonary bypass. PMID- 15550015 TI - Diagnostic score for heparin-induced thrombocytopenia after cardiopulmonary bypass. AB - Heparin-induced thrombocytopenia (HIT) occurs in nearly 3% of patients treated with heparin after cardiopulmonary bypass (CPB). HIT carries a risk of severe thrombotic complications, and must be diagnosed rapidly. To identify simple criteria for estimating the probability of HIT after CPB, we retrospectively analyzed the files of 84 patients with suspected HIT after CPB and we analyzed the usefulness of several variables collected at the time of HIT suspicion to estimate HIT probability. HIT was confirmed in 35 cases and ruled out in 49 cases, on the basis of a platelet increment after heparin withdrawal, detection of heparin-dependent antibodies, and absence of other clear cause of thrombocytopenia. A biphasic platelet count from CPB to the first day of suspected HIT, an interval of >/= 5 days from CPB to the first day of suspected HIT, and a CPB duration of /= 4 days, and surgical patients are included. RESULTS: Of 6160 patients enrolled up to December 2003, 756 (12%) were acutely ill medical patients with immobility >/= 4 days, and 884 (14%) were surgical patients who developed VTE within 2 months of surgical intervention. Only 28% of acutely ill medical patients had received thromboprophylaxis, compared with 67% of surgical patients. During the 3-month follow-up period, both fatal pulmonary embolism (PE) and fatal bleeding occurred more frequently in acutely ill medical patients. Immobility in acutely ill medical patients, cancer, and PE were associated with a significantly higher risk of fatal PE or bleeding. CONCLUSIONS: In patients treated for VTE, the incidences of fatal PE, fatal bleeding, and major bleeding were significantly higher in acutely ill medical patients compared with surgical patients. Given the low percentage of acutely ill medical patients who had received thromboprophylaxis, increasing its use appropriately may reduce the incidence of VTE and associated complications. PMID- 15550018 TI - Divergent regional opinions on thromboembolic complications and prophylaxis among surgeons. PMID- 15550019 TI - A survey of thrombosis prophylaxis use in patients undergoing arthroscopic surgery. PMID- 15550020 TI - The no-reflow phenomenon in coronary arteries. AB - No reflow occurs when there is inadequate myocardial perfusion of a given segment of the coronary circulation without evidence of epicardial vessel obstruction. It is a rare but clinically significant condition associated with myocardial infarction and coronary interventions. Diagnosis is usually based on clinical signs of myocardial ischemia (symptoms and/or ECG changes) combined with coronary angiography. Management can be difficult and primarily consists of intracoronary administration of vasodilators. One interesting etiology is thromboembolism and this has become the focus for new potential treatments, including distal embolic protection devices. PMID- 15550021 TI - T cell recognition of the A2 domain of coagulation factor VIII in hemophilia patients and healthy subjects. AB - Hemophilia A patients treated with coagulation factor VIII (FVIII), and also some healthy subjects, may develop anti-FVIII antibodies (Ab), whose synthesis is driven by FVIII-specific CD4+ T cells. Some Ab block the procoagulant function of FVIII (inhibitors). Many inhibitors recognize epitopes on the FVIII A2 domain. Here, we have sought to identify A2 epitopes recognized by CD4+ T cells. We tested the proliferative response of CD4+ blood lymphocytes (BL) from hemophilia patients and healthy subjects, to overlapping synthetic peptides spanning the A2 domain sequence. Many A2 peptides induced proliferative responses of CD4+ BL from one or more subjects. The peptide-induced responses were strongest in hemophilia patients with inhibitors, weakest in healthy subjects. A2 peptides comprising residues 371-400, 621-650 and 671-690 elicited frequent and strong responses in hemophilia A patients, and especially in those with inhibitors. Healthy subjects recognized frequently only the sequence 371-400. A three-dimensional model of the A2 domain suggests that these CD4+ epitope sequences have structural features typical of 'universal' CD4+ T epitopes. PMID- 15550022 TI - Co-segregation of the PROS1 locus and protein S deficiency in families having no detectable mutations in PROS1. AB - Inherited deficiency of protein S constitutes an important risk factor of venous thrombosis. Many reports have demonstrated that causative mutations in the protein S gene are found only in approximately 50% of the cases with protein S deficiency. It is uncertain whether the protein S gene is causative in all cases of protein S deficiency or if other genes are involved in cases where no mutation is identified. The aim of the current study was to determine whether haplotypes of the protein S gene cosegregate with the disease phenotype in cases where no mutations have been found. Eight protein S-deficient families comprising 115 individuals where previous DNA sequencing had failed to detect any causative mutations were analyzed using four microsatellite markers in the protein S gene region. Co-segregation between microsatellite haplotypes and protein S deficiency was found in seven of the investigated families, one family being uninformative. This suggests that the causative genetic defects are located in or close to the protein S gene in a majority of such cases where no mutations have been found. PMID- 15550023 TI - Treatment effects of drotrecogin alfa (activated) in patients with severe sepsis with or without overt disseminated intravascular coagulation. AB - Disseminated intravascular coagulation (DIC) is a serious condition associated with sepsis. Clinical management of DIC is hampered by lack of clear diagnostic criteria. The International Society on Thrombosis and Haemostasis (ISTH) has proposed a diagnostic scoring algorithm for overt DIC based on routine laboratory tests. The objective was to assess a modified version of the ISTH scoring system and determine the effect of drotrecogin alfa (activated) (DrotAA, recombinant human activated protein C) on patients with DIC. The large database from the PROWESS clinical trial in severe sepsis was retrospectively used to assess a modified ISTH scoring system. Baseline characteristics and treatment effects of DrotAA were evaluated. At baseline, 29% (454/1568) of patients had overt DIC. Overt DIC was a strong predictor of mortality, independent of APACHE II score and age. Placebo-treated patients with overt DIC had higher mortality than patients without (43 vs. 27%). DrotAA-treated patients with overt DIC had a trend towards greater relative risk reduction in mortality than patients without (29 vs. 18%, P = 0.261) but both groups had greater relative risk reduction than placebo-treated patients. Serious bleeding rates during DrotAA infusion in patients with and without overt DIC were slightly increased (P = 0.498), compared with placebo, while clinically overt thrombotic events during the 28-day period were slightly reduced (P = 0.144). Modified ISTH overt DIC scoring may be useful as an independent assessment for identifying severe sepsis patients at high risk of death with a favorable risk/benefit profile for DrotAA treatment. Patients without overt DIC also received significant treatment benefit. PMID- 15550024 TI - A randomized comparison of the effects of aspirin and clopidogrel on thrombotic risk factors and C-reactive protein following myocardial infarction: the CADET trial. AB - A randomized, double-blind multicenter trial-the Clopidogrel and Aspirin: Determination of the Effects on Thrombogenicity (CADET) trial-was carried out to compare the effects of clopidogrel vs. aspirin on thrombotic variables and C reactive protein (CRP), over a 6-month period of treatment, in patients with an acute myocardial infarction within the previous 3-7 days, who were not scheduled for major surgery including coronary artery bypass grafting. Patients (n = 184) were randomly allocated to aspirin (75 mg day(-1)) or clopidogrel (75 mg day( 1)). Blood samples were taken at baseline and then at clinic visits at 1, 3 and 6 months. By 1 month, clottable and immunonephelometric fibrinogen, D-dimer, von Willebrand factor, factor VIII and CRP were significantly (P < 0.05) reduced from baseline values in both treatment groups; as well as tissue plasminogen activator antigen in the aspirin group only. At 6 months, there were no differences between treatment groups (P > 0.05) for any of the variables, whether or not potential confounding variables were adjusted for. Similarly, there were no differences between treatments in the difference between baseline and final values for any of the variables. Aspirin and clopidogrel were thus found to have similar effects on thrombotic variables and CRP in this patient population. PMID- 15550025 TI - Severe hemophilia A due to a 1.3 kb factor VIII gene deletion including exon 24: homologous recombination between 41 bp within an Alu repeat sequence in introns 23 and 24. AB - Partial or complete factor (F)VIII gene deletions are found in about 5% of families with severe hemophilia A. Relatively few deletions have been well characterized and, of these, recombination occurred between either common repeat elements or non-homologous sequences. In evaluating a family with severe hemophilia A, an exon 24 deletion was suspected when no fragment was obtained on attempted PCR amplifications. A combination of the 5' primer of exon 23 and the 3' primer of exon 25 fragments was used with prolonged extension times to amplify a normal 2.9 kb fragment that included exons 23 through 25; the patient's amplified product was 1.6 kb indicating a 1.3 kb deletion. A mixture of normal and patient DNA showed both sized fragments as did that from an obligate carrier. Carrier detection was applied to two women at risk; one was and one was not a carrier. Sequencing the proband's 1.6 kb fragment revealed that a 1328 bp deletion occurred between homologous sequences of 287 and 285 bp in introns 23 and 24, respectively; these share 85% identity. Blast nucleotide search revealed that these represent Alu elements. Comparison with an alignment of each of the two homologous sequences further localized recombination to a 41-bp segment. However, a simple recombination event would not account for the proband's sequence. The most likely explanation is that the homologous recombination was accompanied by incomplete mismatch repair. PMID- 15550026 TI - Guidelines on preparation, certification, and use of certified plasmas for ISI calibration and INR determination. AB - Reliable international normalized ratio (INR) determination depends on accurate values for international sensitivity index (ISI) and mean normal prothrombin time (MNPT). Local ISI calibration can be performed to obtain reliable INR. Alternatively, the laboratory may determine INR directly from a line relating local log(prothrombin time [PT]) to log(INR). This can be done by means of lyophilized or frozen plasmas to which certified values of PT or INR have been assigned. Currently there is one procedure for local calibration with certified plasmas which is a modification of the WHO method of ISI determination. In the other procedure, named 'direct' INR determination, certified plasmas are used to calculate a line relating log(PT) to log(INR). The number of certified plasmas for each procedure depends on the method of preparation and type of plasma. Lyophilization of plasma may induce variable effects on the INR, the magnitude of which depends on the type of thromboplastin used. Consequently, the manufacturer or supplier of certified plasmas must assign the values for different (reference) thromboplastins and validate the procedure for reliable ISI calibration or 'direct' INR determination. Certification of plasmas should be performed by at least three laboratories. Multiple values should be assigned if the differences between thromboplastin systems are greater than 10%. Testing of certified plasmas for ISI calibration may be performed in quadruplicate in the same working session. It is recommended to repeat the measurements on three sessions or days to control day-to-day variation. Testing of certified plasmas for 'direct' INR determination should be performed in at least three sessions or days. Correlation lines for ISI calibration and for 'direct' INR determination should be calculated by means of orthogonal regression. Quality assessment of the INR with certified plasmas should be performed regularly and should be repeated whenever there is a change in reagent batch or in instrument. Discrepant results obtained by users of certified plasmas should be reported to manufacturers or suppliers. PMID- 15550027 TI - Clinical measurement of thrombin generation by calibrated automated thrombography requires contact factor inhibition. AB - BACKGROUND: Measurement of thrombin generation by calibrated automated thrombography (CAT) could fulfill the requirements of a global test of coagulability and is potentially applicable to routine clinical laboratory practice. The purpose of this study was to determine if corn trypsin inhibitor (CTI) could be used to abolish contact factor activation in this assay, thus allowing accurate measurement of low tissue factor (TF) concentration-triggered thrombin generation on samples taken in a routine clinical setting. METHODS: The endogenous thrombin potential (ETP) was measured by CAT. RESULTS: The study demonstrated that addition of CTI after plasma separation is not sufficient and blood must be drawn into tubes containing CTI if in-vitro contact factor activated thrombin generation is to be abolished. Contact factor-activated thrombin generation is completely inhibited at a CTI concentration of 18.3 microg mL(-1) whole blood. Increasing the CTI concentration above this level does not lead to suppression of the TF-triggered ETP. At a TF concentration of 2 pmol, ETPs were significantly lower in the presence of CTI (P < 0.001). The difference (no CTI minus CTI) between results ranged from - 1 to 2159 nM min(-1) (median - 754). Whilst the low concentration TF-ETP assay was not optimized to distinguish degrees of coagulability between patient samples, there was a significant difference in ETP between normal and hemophilia samples and samples from patients with a clinical prothrombotic tendency. CONCLUSIONS: CTI can be applied to ETP measurement by CAT. This permits the use of CAT in a low TF-triggered thrombin generation assay without concern for the effect of interference from in-vitro contact factor activation and the optimum reagent conditions for using CAT as a global test of coagulability in clinical practice can now be defined. PMID- 15550028 TI - Regulation of local availability of active tissue-type plasminogen activator in vivo in man. AB - Free, biologically active tissue-type plasminogen activator (tPA) is the main initiator of intravascular fibrinolysis, but little is known about the regulation of active tPA on the organ level. The aim was to investigate if the local availability of active tPA on the organ level depends on the local release rate of tPA or the arterial input of tPA and plasminogen activator inhibitor type 1 (PAI-1). Also, we wanted to evaluate if plasma levels predict capacity for endothelial release of fibrinolytic proteins. Invasive perfused-forearm studies were performed in 96 healthy subjects. Local release rates of fibrinolytic proteins were assessed at baseline and during endothelial stimulation. Stimulation by methacholine and desmopressin induced a 6- and 12-fold increase in total tPA release rates, respectively. With increasing local release rates of tPA a gradually closer correlation emerged between the total tPA secretion and the forearm output of active tPA (from r = 0.102, ns to r = 0.85, P < 0.0001). Forearm availability of active tPA was not related to arterial input of either tPA or PAI-1. Release rates and plasma levels of tPA were not correlated. Baseline release rates of active tPA increased to noon. The major determinant for the local availability of active tPA is the capacity of the endothelium to release tPA rather than the arterial input of PAI-1 or tPA. Despite a molar excess of PAI-1, the majority of tPA released during stimulation does not undergo local inactivation. The capacity to release tPA locally cannot be predicted from its plasma concentration. PMID- 15550029 TI - Role of mRNA transcript stability in modulation of expression of the gene encoding thrombin activable fibrinolysis inhibitor. AB - Regulation of mRNA stability has emerged as a major control point in eukaryotic gene expression. The abundance of a particular mRNA can be rapidly regulated in response to a stimulus by altering the stability of existing translatable transcripts rather than by altering the rate of transcription initiation. Alternative polyadenylation of transcripts during mRNA processing can be important in determining transcript abundance if the different forms of mRNA possess different stabilities or translatability. The mRNA transcript encoding thrombin activable fibrinolysis inhibitor (TAFI) is an attractive candidate for regulation of mRNA stability because of the relatively long length of its 3' untranslated region and because the transcript can be polyadenylated at three different sites. As well, we have previously reported that treatment of HepG2 cells with interleukins (IL) - 1beta and - 6 destabilizes the endogenous TAFI mRNA expressed in this cell line. In the current study, we report that the TAFI 3'-untranslated region contains cis-acting instability element(s) and that these elements in fact determine the intrinsic stability of the TAFI transcript. Moreover, we found that the three different polyadenylated mRNA forms have different intrinsic stabilities, with the mRNA half-life increasing from the longest to the shortest transcript. Interestingly, treatment with IL-1beta plus IL-6 not only resulted in a 2-fold decrease in stability of the transcript produced using the 3'-most polyadenylation site but also resulted in profound shifts in the relative abundances of the respective polyadenylated forms through changes in the frequency of utilization of the three polyadenylation sites. As such, in the presence of IL-1beta and IL-6, the longest transcript is over a thousand times more abundant than the two shorter transcripts whereas in the absence of the stimulus it comprises only 1% of the total TAFI transcripts. PMID- 15550030 TI - Adenine triphosphate nucleotides are antagonists at the P2Y receptor. AB - The aim of the present study was to characterize the pharmacological profile of the P2Y(12) receptor for several adenine triphosphate nucleotides in view of their possible roles as partial agonists or true antagonists. Two distinct cellular systems were used: P2Y(1) receptor deficient mouse platelets ( platelets) previously shown to express a native and functional P2Y(12) receptor and 1321 N1 astrocytoma cells stably expressing the human P2Y(12) receptor (1321 N1 P2Y(12)). ADP and its structural analogues inhibited cAMP accumulation in a dose-dependent manner in both platelets and 1321 N1 P2Y(12) cells with a similar rank order of potency, 2 methylthio-ADP (2MeSADP) >>ADP - Adenosine 5'-(betathio) diphosphate (AlphaDPbetaS). Commercial ATP, 2 chloro; ATP (2ClATP) and 2 methylthio-ATP (2MeSATP) also inhibited cAMP accumulation in both cell systems. In contrast, after creatine phosphate (CP)/creatine phosphokinase (CPK) regeneration, adenine triphosphate nucleotides lost their agonistic effect on platelets and behaved as antagonists of ADP (0.5 microm)-induced adenylyl cyclase inhibition with IC(50) of 13.5 +/- 4.8, 838 +/- 610, 1280 +/- 1246 microm for 2MeSATP, ATP and 2ClATP, respectively. In 1321 N1 P2Y(12) cells, CP/CPK regenerated ATP and 2ClATP lost their agonistic effect only when CP/CPK was maintained during the cAMP assay. The stable ATP analogue ATPgammaS antagonized ADPbetaS-induced inhibition of cAMP accumulation in both platelets and 1321 N1 P2Y(12) cells. Thus, ATP and its triphosphate analogues are not agonists but rather antagonists at the P2Y(12) receptor expressed in platelets or transfected cells, provided care is taken to remove diphosphate contaminants and to prevent the generation of diphosphate nucleotide derivatives by cell ectonucleotidases. PMID- 15550031 TI - Targeting platelet GPIbalpha transgene expression to human megakaryocytes and forming a complete complex with endogenous GPIbbeta and GPIX. AB - Bernard-Soulier Syndrome (BSS) is a severe congenital platelet disorder that results from a deficiency of the platelet membrane glycoprotein (GP) Ib/IX complex that is composed of four subunits (GPIbalpha, GPIbbeta, GPIX, and GPV). Mutations in either GPIbalpha, GPIbbeta, or GPIX can result in BSS with many of the known mutations occurring in GPIbalpha. In this study, we have developed a gene therapy strategy to express hemagglutinin (HA)-tagged GPIbalpha in megakaryocytes and potentially correct a hereditary deficiency. To direct GPIbalpha expression in megakaryocytic lineage cells, we designed a GPIbalpha cassette where human GPIbalpha cDNA was placed under control of the megakaryocytic/platelet-specific alphaIIb promoter and inserted into a lentiviral vector. Human CD34+ peripheral blood cells (PBC) and Dami cells were transduced with alphaIIb-HA-GPIbalpha-WPT virus. Flow cytometry analysis demonstrated that 50.1% of the megakaryocytes derived from CD34+ stem cells and 97.3% of Dami cells were transduced and expressed transgene GPIbalpha protein. Immunoprecipitation with Western blot analysis demonstrated that transgene protein associated with endogenous GPIbbeta and GPIX proteins. To address further the lineage-specific expression of the alphaIIb-HA-GPIbalpha construct, three cell lines, Dami, AtT-20 and HepG2, were transfected with GPIbalpha expression plasmids and analyzed by confocal microscopy. The results demonstrated that among these three cell lines, the tissue-specific alphaIIb promoter was active only in Dami cells. Thus, GPIbalpha can be efficiently and specifically expressed in the megakaryocytic compartment of hematopoietic cells and the transgene product associates with endogenous GPIbbeta and GPIX forming a complete complex. This strategy could potentially be utilized for gene therapy of BSS. PMID- 15550032 TI - Cleavage of platelet endothelial cell adhesion molecule-1 (PECAM-1) in platelets exposed to high shear stress. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1, CD31) is a 130 kDa transmembrane glycoprotein that belongs to the immunoglobulin superfamily and is expressed on the surface of endothelial cells, platelets, and other blood cells. Although the importance of this adhesion molecule in various cell-cell interactions is established, its functional role in platelets remains to be elucidated. In this study, we examined whether PECAM-1 underwent changes in platelets exposed to high shear stress. Platelet PECAM-1 was cleaved under high shear stress and was released into the extracellular fluid as a fragment with an approximate molecular weight of 118 kDa. The cleavage was inhibited by an anti VWF MoAb, but not by recombinant VWF A1 domains. These findings suggest that the GPIb-VWF interaction is involved in PECAM-1 cleavage under high shear stress, and that the cleavage is independent of GPIb clustering by VWF multimers. Furthermore, EGTA or calpeptin inhibited PECAM-1 cleavage. This finding provides evidence for the involvement of calpain in PECAM-1 cleavage. Flow-cytometric analysis revealed that PECAM-1 expression on the platelet surface was decreased under high shear stress. This reduction occurred exclusively in a specific population of platelets, which corresponded to platelet-derived microparticles (PMP). In conclusion, PECAM-1 cleavage under high shear stress is closely related to the activation of calpain and the process of PMP formation mediated by the GPIb-VWF interaction. PMID- 15550033 TI - Hemostatic and hematological abnormalities in gain-of-function fps/fes transgenic mice are associated with the angiogenic phenotype. AB - The Fps/Fes tyrosine kinase has been implicated in the regulation of hematopoiesis and inflammation. Mice expressing an activated variant of Fps/Fes (MFps) encoded by a gain-of-function mutant transgenic fps/fes allele (fps(MF)) exhibited hematological phenotypes, which suggested that Fps/Fes can direct hematopoietic lineage output. These mice also displayed marked hypervascularity and multifocal-hemangiomas which implicated this kinase in the regulation of angiogenesis. Here we explored the potential involvement of Fps/Fes in the regulation of hemostasis through effects on blood cells and the vascular endothelium. Hematological parameters of fps(MF) mice were characterized by peripheral blood analysis, histology, and transmission electron microscopy. Hemostasis parameters and platelet functions were assessed by flow cytometry and measurements of activated partial thromboplastin time, prothrombin time, thrombin clot time, platelet aggregation, bleeding times and in vitro fibrinolytic assays. Hematological and morphological analyses showed that fps(MF) mice displayed mild thrombocytopenia, anemia, red cell abnormalities and numerous hemostatic defects, including hypofibrinogenemia, hyper-fibrinolysis, impaired whole blood aggregation and a mild bleeding diathesis. fps(MF) mice displayed a complex array of hemostatic perturbations which are reminiscent of hemostatic disorders such as disseminated intravascular coagulation (DIC) and of hemangioma-associated pathologies such as Kasabach-Merritt phenomenon (KMS). These studies suggest that Fps/Fes influences both angiogenic and hemostatic function through regulatory effects on the endothelium. PMID- 15550034 TI - PECAM-1 (CD31) is required for interactions of platelets with endothelial cells after irradiation. AB - Sustained adhesion of platelets to endothelial cells (EC) is believed to contribute to thrombosis and vascular occlusions following radiation exposure leading to organ functional impairment and even death. Our objective was to evaluate the role of platelet endothelial cell adhesion molecule (PECAM)-1 in the prothrombotic response of EC after irradiation. Endothelial PECAM-1 expression was determined by cell-enzyme linked immunosorbent assay (ELISA) on human microvascular EC from lung (HMVEC-L) up to 21 days after a 10 Gy irradiation. Platelet- and leukocyte-endothelial cell interactions were assessed using a flow adhesion assay with fluorescently labeled whole blood, and the function of PECAM 1 in these processes was measured by using blocking antibody. PECAM-1 expression was significantly increased on irradiated HMVEC-L and remained elevated at 21 days. Anti-PECAM-1 antibody significantly inhibited adhesion of single platelets and thrombi on irradiated HMVEC-L. This inhibitory effect persisted at day 21. Anti-PECAM-1 also reduced leukocyte adhesion to irradiated HMVEC-L. The up regulation of endothelial PECAM-1 following radiation exposure is persistent. PECAM-1 plays a key role platelet adhesion/aggregation on irradiated EC. Therefore, strategies targeting this adhesion molecule may prevent the development of radiation pathologies. PMID- 15550035 TI - Hirudin ameliorates intestinal radiation toxicity in the rat: support for thrombin inhibition as strategy to minimize side-effects after radiation therapy and as countermeasure against radiation exposure. AB - BACKGROUND: The small bowel is a dose-limiting normal tissue in radiation therapy of malignancies in the abdomen and pelvis, as well as an important determinant of survival after non-therapeutic radiation exposure. Irradiation of normal tissues, including intestine, causes loss of vascular thromboresistance and upregulation of thrombin receptors. Radiation-induced endothelial dysfunction is thought to be involved in both early and delayed radiation responses. Hence, thrombin may be a potential target for ameliorating normal tissue radiation toxicity. OBJECTIVE: To assess direct thrombin inhibition as a protective strategy against small bowel radiation toxicity. METHODS: Rat small intestine was exposed to localized orthovoltage X-radiation. Recombinant hirudin, a direct thrombin inhibitor, or vehicle was infused from 2 days before irradiation to 14 days after irradiation. Structural, cellular, and molecular aspects of intestinal radiation injury were assessed at 2 weeks (early toxicity) and 26 weeks (chronic toxicity) after irradiation. RESULTS: Compared with unirradiated intestine, irradiated intestine showed increased expression of tissue factor, increased immunoreactivity for enzymatically active thrombin, and increased extravascular fibrin(ogen) deposition. Hirudin treatment significantly attenuated radiation-induced mucosal damage (P = 0.04), reactive intestinal wall thickening (P = 0.02), transforming growth factor-beta immunoreactivity levels (P = 0.0002), and collagen III deposition (P = 0.003). The differences between hirudin-treated and control rats were more pronounced at 2 weeks than at 26 weeks after irradiation. Hirudin treatment did not affect postradiation granulocyte infiltration. CONCLUSIONS: Short-term thrombin inhibition attenuates important aspects of intestinal radiation toxicity. Thrombin is a promising target for minimizing normal tissue injury after radiation therapy of cancer, as well as for protecting normal tissues from the adverse effects of non-therapeutic radiation exposure. PMID- 15550036 TI - Prevention of venous thromboembolism after orthopedic surgery: the EXPRESS study. PMID- 15550039 TI - Wine, alcohol and cardiovascular risk: open issue. PMID- 15550044 TI - Experimental bases of the vascular protective effect of wine: studies on Lambrusco samples. PMID- 15550045 TI - Protein Z plasma levels are not associated with the risk of coronary heart disease: the PRIME Study. PMID- 15550046 TI - Aspirin resistance in vitro and hypertension in stroke patients. PMID- 15550047 TI - Measurement of plasma von Willebrand factor in acute coronary syndromes and the influence of ABO blood group status. PMID- 15550048 TI - Type 2B von Willebrand disease and related disorders of patients with increased ristocetin-induced platelet aggregation: what they tell us about the role of von Willebrand factor in hemostasis. PMID- 15550049 TI - Type 2 A (group II) von Willebrand disease mutations increase the susceptibility of VWF to ADAMTS-13. PMID- 15550050 TI - Overall hemostatic potential (OHP) assay-a possible tool for determination of prothrombotic pattern in FXII deficiency. PMID- 15550051 TI - Prothrombotic changes in users of combined oral contraceptives containing drospirenone and cyproterone acetate. PMID- 15550052 TI - Thalidomide, deep venous thrombosis and vasculitis. PMID- 15550053 TI - Thalidomide in front line treatment in multiple myeloma: serious risk of venous thromboembolism and evidence for thromboprophylaxis. PMID- 15550054 TI - Shedding of tissue factor (TF)-containing microparticles rather than alternatively spliced TF is the main source of TF activity released from human cancer cells. PMID- 15550055 TI - Measurement of soluble P-selectin and soluble CD40 ligand in serum and plasma. PMID- 15550056 TI - DNA extraction yield is associated with several phenotypic characteristics: results from two large population surveys. PMID- 15550057 TI - In vivo thrombin generation in neonates. PMID- 15550058 TI - Factor IX polypyrimidine tract mutation analysis using mRNA from peripheral blood leukocytes. PMID- 15550059 TI - Kinetic spectrophotometric factor XIII activity assays: the subtraction of plasma blank is not omissible [corrected]. PMID- 15550060 TI - The safety of recombinant factor VIIa: a rebuttal. PMID- 15550064 TI - Antibodies to Chlamydia trachomatis heat shock proteins Hsp60 and Hsp10 and subfertility in general population at age 31. AB - PROBLEM: To assess the association between antibodies to Chlamydia trachomatis heat shock proteins 60 and 10 (Hsp60 and Hsp10) and subfertility in a general population sample. METHOD OF STUDY: A nested case (n = 146)-control (n = 278) study in a population-based birth cohort. Serum immunoglobulin (Ig)G and IgA antibodies against C. trachomatis Hsp60 and Hsp10, explanatory factors, were measured by enzyme immunoassay, using recombinant proteins as antigens. The main outcome variable was subfertility (time to pregnancy > or =12 months). RESULTS: The prevalence and medians of serum IgA antibodies to Hsp60 and Hsp10 were significantly higher in the female partners of subfertile couple than in their fertile controls. On the contrary, among male partners of subfertile couple, especially among smokers serum antibody levels to Hsp antigens were lower than in the controls. CONCLUSION: The results indicate a serological association of antibodies to chlamydial Hsp antigens with female subfertility in a population based sample. PMID- 15550065 TI - Identification of the M-CSF receptor in endometriosis by immunohistochemistry and RT-PCR. AB - PROBLEM: The aim of this paper is to provide further evidence that the dystopic proliferation of endometriotic epithelia is caused by the stimulation of peritoneal macrophages. It is essential to show that endometriotic epithelial cells express the macrophage colony-stimulating factor receptor (M-CSFR) which binds the M-CSF produced by the peritoneal macrophages. METHOD OF STUDY: For the detection of M-CSFR, samples of ectopic endometrium (n = 79) and eutopic endometrium (n = 18) were compared. The specimens were gained at operative laparoscopy in the proliferative phase of the cycle. Cryostat sections were used for immunohistochemical detection. For in vitro reverse transcriptase polymerase chain reaction (RT-PCR) tests, the tissue was immediately shock frozen on paraffin sections. For the in situ RT-PCR technique the specimens were placed in a para-formaldehyde solution, embedded in paraffin and later processed. The Gene Amp 1000 in situ PCR system (Perkin Elmer) was used as the thermal cycler. RESULTS: M-CSF and the M-CSF receptor are present in eutopic and ectopic endometrium. Qualitatively, with both PCR techniques we found the M-CSF receptor to be present in all samples examined. Using the histochemical detection technique, the M-CSF receptor was found in nearly 70% of endometriosis patients compared with a statistically significant lower percentage in normal endometrium. CONCLUSIONS: The in situ RT-PCR technique and immunohistochemistry elaborated the need to trace the cellular sources of the M-CSF receptor. The identification of the M-CSF receptor in endometriotic tissue and in endometrium is apt to open a new experimental field in endometriosis research. PMID- 15550066 TI - Possible pathophysiological roles of mitogen-activated protein kinases (MAPKs) in endometriosis. AB - PROBLEM: Endometriosis accompanies local inflammatory reactions in the peritoneal cavity. We examined the phosphorylation of mitogen-activated protein kinases (MAPKs), i.e. extracellular signal-regulated kinase (ERK), p38 MAPK (p38) and c Jun N-terminal kinase (JNK) in endometriotic stromal cells, and their possible pathophysiological roles in endometriosis in relation to proinflammatory substances. METHOD OF STUDY: Endometriotic stromal cells were isolated from endometriomas and were cultured for the experiments. Phosphorylation of MAPKs in endometriotic stromal cells treated with interleukin (IL)-1beta, tumor necrosis factor (TNF)alpha and H(2)O(2) were examined by Western blot analysis. Effects of PD98059, SB202190 and SP600125 (inhibitors of ERK, p38 and JNK, respectively) on IL-1beta-induced secretion of IL-6 and IL-8, and on IL-1beta-induced expression of cyclo-oxygenase-2 (COX-2) in endometriotic cells were studied. In addition, eutopic endometrial tissues were collected, and the phosphorylation rate of p38 in eutopic endometrial tissues and endometriotic tissues were determined. RESULTS: IL-1beta, TNFalpha and H(2)O(2) stimulated the phosphorylation of ERK, p38 and JNK, while the total amounts of proteins of the respective MAPKs were virtually the same compared with those in the unstimulated controls. Both SB202190 and SP600125 suppressed IL-1beta-induced secretion of IL-6 and IL-8, and PD98059 suppressed IL-1beta-induced secretion of IL-8. Both SB202190 and PD98059 suppressed IL-1beta-induced expression of COX-2 in endometriotic cells. The p38 phosphorylation rates in the endometriotic tissues were significantly higher than those in the eutopic endometrial tissues of the same patients. CONCLUSIONS: Given the current theory that inflammatory changes are involved in the progression of endometriosis, MAPKs could play as pivotal intracellular signal transducers in endometriotic cells, and thus have a pathophysiological role in the disease. PMID- 15550067 TI - Influence of thyroid autoimmunity and maternal age on the risk of miscarriage. AB - OBJECTIVES: Recently, studies have shown an association between antiperoxidase for the detection of thyroid autoimmunity (TAI) and abortion. Another point to be considered is the association of high risk of abortion and maternal age. The aim of the present study was to evaluate if the association between TAI and miscarriage could also be verified whether a population of unselected pregnant young women who normally present a low risk of miscarriage. MATERIALS AND METHODS: We studied 534 pregnant women, by determining their serum thyroid antiperoxidase antibodies (TPO-Abs), thyrotropin (TSH) and free thyroxine (fT4) levels. Our end point was the pregnancy loss or live birth. RESULTS: Age ranged from 12 to 49 years (mean +/- S.D.; 23.5 +/- 5.9). Of 534 women, 29 (5.4%) were TPO-Ab positive. TSH levels were significantly higher in TPO-Ab-positive women compared with TPO-Ab negative women (median; 1.9 versus 1.1; P = 0.001). Elevated TSH levels were found in 13.8% (4 of 29) of the TPO-Ab-positive women compared with only 2.4% (12 of 505) in the TPO-Ab-negative women. There were no significant differences in fT4 levels in relation with autoimmunity and risk of miscarriage. The overall risk of miscarriage was 2.4% (13 of 534). Risk of miscarriage was significantly higher among women older than 35 years (7.7%), TPO Ab positive (10.3%) and presenting high levels of TSH (12.5%). These factors remained independently associated with the risk of miscarriage in full multivariate analysis. CONCLUSIONS: We conclude that TAI is independently associated with is a higher risk of miscarriage in a population of unselected pregnant presenting a low risk of miscarriage. PMID- 15550068 TI - Effect of proteasome pathway on initiation of mouse labor induced by antiprogesterone. AB - PROBLEM: Various kinds of contraction-associated molecules are up-regulated at the initiation of labor. However, expression profiling has revealed that many molecules are also down-regulated. The effect of down-regulation of molecules by protein degradation on parturition is not known. METHODS OF STUDY: We administered lactacystin, a specific proteasome inhibitor, to mouse preterm birth model induced by antiprogesterone RU486 on day 16.0 post-coitus. NF-kappaB activity, and the levels of transcripts for oxytocin receptor, prostaglandin F(2alpha) receptor (FP), cyclooxygenase-1, -2, and interleukin-1beta in the uterus were examined by electrophoretic mobility shift assay and semi quantitative reverse transcriptase-polymerase chain reaction, respectively. RESULTS: Administration of lactacystin significantly prolonged the time until the delivery of the first pup. FP mRNA level was solely elevated by RU486 treatment, and lactacystin significantly suppressed this up-regulation. CONCLUSIONS: Proteolysis by proteasomes in the uterus regulates the initiation of labor, at least in part, via control of contraction-associated molecules such as FP. PMID- 15550069 TI - CD11b Expression on circulating leukocytes increases in preparation for parturition. AB - PROBLEM: Initiation of parturition is associated with migration of leukocytes to the reproductive tract. This migration is controlled in part by expression of adhesion molecules on the surface of leukocytes and vascular endothelial cells. Within the reproductive tract, certain endothelial adhesion molecules, including intercellular adhesion molecule-1 (ICAM-1), are up-regulated at the end of gestation. ICAM-1 binds to the beta(2) integrin CD11b on the leukocytes. In this study, we wanted to investigate whether complementary changes occur in expression of adhesion molecules on maternal leukocytes in preparation for parturition. METHOD OF STUDY: We used flow cytometry to ascertain changes in adhesion molecules expression on leukocytes throughout third trimester and labor. RESULTS: We found a significant increase in the expression of CD11b on monocytes and granulocytes in women at >37 weeks of gestation. CONCLUSIONS: CD11b may be a key molecule for leukocyte trafficking to the reproductive tract at the end of pregnancy. PMID- 15550070 TI - Antichlamydial and antisperm antibodies in patients with chlamydial infections. AB - PROBLEM: Establishing the correlation between antichlamydial antibodies (AchAbs) and antisperm antibodies (ASA) in patients with chlamydial infections. METHOD OF STUDY: ASA were studied in sera from patients (142 with genital, 57 with ocular chlamydial infections) and control group (n = 100) by gelatin and tray agglutination test (TAT), sperm immobilization test (SIT) and ELISA. AchAbs were revealed by ELISA. RESULTS: A significantly higher (P < 0.05) ASA incidence was noted in patients with genital infections as compared with controls and patients with ophthalmologic infection (P < 0.0001), but not between patients with ophthalmologic infection and controls (P > 0.05). A significant correlation was established between AchAbs and ASA for TAT (r = 0.8214, P = 0.0341), SIT (r = 0.797, P = 0.032) and ELISA (r = 0.8519, P = 0.0313) in patients with genital infections only. CONCLUSIONS: The genital Chlamydia infection may play a role in the induction of ASA. This is probably a result of the inflammatory process, but not of cross-reactivity between sperm and Chlamydia trachomatis antigens. PMID- 15550071 TI - A response to 'Shall we properly re-examine the status of allogeneic lymphocyte therapy for recurrent early pregnancy failure?', Clark D A. Am J Reprod Immunol 2004; 51:7-15. PMID- 15550076 TI - Abdominal aortic aneurysm repair: balance of risk. PMID- 15550077 TI - Cancer statistics: everything you wanted to know about the cancer registry data but were too afraid to ask. AB - Australia and New Zealand (ANZ) have had complete population cancer incidence registration for decades and are beginning to develop clinical data systems to collect details of staging and treatment. Sadly, this bounty of statistical data is often under utilized and occasionally abused. Many are unaware of the data and statistical services available from their local cancer registry. To promote the use of ANZ population cancer data, the following paper describes the principal statistics that are available and identifies common problems with their use. PMID- 15550078 TI - Four-year follow up of patients with untreated abdominal aortic aneurysms. AB - BACKGROUND: The data in the literature are still controversial describing the outcome of patients not treated for a large abdominal aortic aneurysm (AAA) especially with significant comorbidities. We followed up patients trying to establish their long-term outcome. METHOD: Since 1998, we have prospectively followed all patients referred to our department with AAA. A retrospective analysis was carried out selecting all patients who had an AAA larger than 5 cm, and who declined or were declined for operative repair between February 1998 and November 2001. RESULTS: One hundred and eleven patients were included in the present study. There were 78 men and 33 women. The mean age was 80 years. At the end of the study, 65 patients (59%) were deceased. Ruptured aneurysm occurred in 27 patients (median time to rupture = 14 months) with one patient surviving an emergency repair. Thirty-nine patients died from unrelated illnesses. In the 5 5.9 cm AAA group (n = 58), out of 31 deceased patients, five (16%) have died of ruptured AAA. In the 6 cm and larger AAA group (n = 53), out of 34 deceased patients, 21 (62%) have died of ruptured AAA. There was no significant difference in survival between patients with AAA below and above 6 cm in diameter (P = 0.15). CONCLUSION: In the presence of significant comorbidities, most patients with AAA less than 6 cm died from unrelated illnesses. In the larger AAA group, the likelihood of death from AAA rupture or unrelated illnesses is almost equal. PMID- 15550079 TI - Pancreatic cancer: management and survival. AB - AIM: To review the management and survival from all pancreatic cancer over a 5 year period at a tertiary referral hospital in New Zealand and to examine similar outcome data from the national cancer registry. METHODS: A retrospective audit was conducted for the 5-year period 1994-99 of patients discharged from Christchurch Hospital (Christchurch, New Zealand) and all patients in the New Zealand Cancer Registry with a diagnosis of pancreatic cancer. Kaplan- Meier survival curves were used for analysis. RESULTS: From Christchurch Hospital a total of 230 patients were identified with a discharge diagnosis of pancreatic cancer. Medium survival for all groups was 3.9 months. There was a median survival of 1.6 months for the non-interventional group, 3.1 months for the stent group, 6.2 months for the bypass group and 12.6 months for the pancreatico duodenectomy group. These data are very similar to the New Zealand National Cancer Registry data, where the overall median survival was 3.1 months and median survival for a pancreatico-duodenectomy was 13.9 months. CONCLUSION: A pancreatico-duodenectomy is usually a palliative surgical technique and not a curative procedure. Those selected for resection have been shown to have an advantage over operative bypass in terms of length of survival, however, this most likely reflects selection bias. PMID- 15550080 TI - Haemorrhagic complications of pancreaticoduodenectomy. AB - BACKGROUND: Haemorrhagic complication occurs in 5-16% of patients following pancreaticoduodenectomy (PD). We report an analysis of patients with post-PD bleed, to identify predictors of bleed, predictors of survival following bleed and the management of post-PD bleed. METHODS: Two hundred and eighteen patients with periampullary cancers underwent PD from 1989 to 2002. Forty-four (20.2%) patients had a bleeding complication. Of these, 25 patients had an intra abdominal (IA) bleed and 21 had gastrointestinal (GI) bleed (two had both IA and GI bleed). Clinical, biochemical and tumour characteristics were analysed to identify factors influencing bleeding complications. RESULTS: The median time to presentation was 4.5 days (0-21 days). Serum bilirubin (P = 0.000, OR: 1.090) and pancreaticojejunostomy (PJ) leak (P = 0.009, OR: 3.174) were significant independent factors predicting bleeding complications. Forty-three per cent of patients each had early bleed (<48 h after PD) or delayed bleed (7 days after PD). Comparison of early and late bleeds showed that IA bleed (P = 0.02) presented as early bleeds. Male sex (P = 0.00) longer duration of jaundice (P = 0.02), PJ leak (P = 0.001), HJ leak (P = 0.001), duct to mucosa type of PJ anastomosis (P = 0.03) and IA abscess (P = 0.00) were associated with a significantly higher incidence of late bleeds. Overall mortality after PD was 9.6% with 34% and 3% in bleeders and non-bleeders, respectively. Septicaemia (P = 0.01, OR: 5.49), and acute renal failure (P = 0.01) were associated with increased mortality. CONCLUSIONS: Bleeding complications following PD were seen in one-fifth of patients and were associated with high mortality. Serum bilirubin levels and PJ leak were significant factors associated with bleeding complications. Septicaemia and acute renal failure were significant factors associated with mortality in the bleeders. PMID- 15550081 TI - Audit of postoperative chemoradiotherapy as adjuvant therapy for resected gastroesophageal adenocarcinoma: an Australian multicentre experience. AB - BACKGROUND: Improved disease free and overall survivals were seen in curatively resected patients with gastric and gastroesophageal adenocarcinoma treated with the Intergroup 0116 (INT 0116) protocol of postoperative adjuvant chemoradiotherapy compared to surgery alone. This protocol has not been widely adopted in Australian centres because of perceived risks of toxicity. METHODS: We reviewed the case records from 45 consecutive patients treated between May 1998 and August 2003 with the INT 0116 protocol and variations at five Australian institutions. The median age was 61.5 years (range 38-79). Twenty-nine patients had gastric and 12 had gastroesophageal junction primaries. All patients had attempted curative resection, however, seven had involved microscopic margins (R1 resection). Thirty-five had regional node involvement and none had evidence of distant metastasis. RESULTS: The overall National Cancer Institute-Common Toxicity Criteria (NCI-CTC) version 2.0 grade 3 and grade 4 toxicity rates for all patients were 37.8% and 4.4%, respectively. There were no treatment related deaths. Gastrointestinal grade 3 toxicity was observed in 20% of patients, while haematologic grade 3 and 4 toxicity was observed in 17.8%. Toxicities experienced led to chemotherapy dose reductions in 22 patients and dose delay in 11 patients. Seven patients had a delay in radiotherapy and two did not proceed with radiotherapy. At a median follow up of 16 months (range 5-35) from surgery, 28 patients have relapsed (six with local recurrence alone) with 22 deaths occurring, all but one caused by cancer. CONCLUSION: The INT 0116 protocol is a safe and feasible schedule in a multicentre setting with an acceptable rate of toxicity and is an appropriate adjuvant treatment option for high-risk resected gastroesophageal adenocarcinoma. PMID- 15550082 TI - Oestrogen receptors revisited: long-term follow up of over five thousand breast cancer patients. AB - BACKGROUND: The oestrogen receptor status of a breast tumour predicts the response to hormonal treatment and is an important prognostic marker; women with oestrogen receptor positive tumours having a better short-term survival outcome. METHODS: Kaplan-Meier estimates and Cox proportional hazard model were used to estimate the association between oestrogen receptor levels and long-term breast cancer-specific survival outcomes in 5735 women diagnosed with breast carcinoma from 1970 to 1997 in Western Australia. Further analysis was performed on a subset of women for whom biochemical and tumour characteristics were also available. RESULTS: Five-year breast cancer-specific survival estimates for women with oestrogen receptor positive tumours was 0.85 (95% CI 0.84-86) compared to 0.72 (95% CI 0.70-74) for women with oestrogen receptor negative tumours. The relative survival advantage of an oestrogen positive tumour over oestrogen negative tumours disappeared by the fourth year (0.8, 95% CI 0.6-1.0). Conditional upon surviving 5 years, long-term breast cancer-specific survival was better for women with oestrogen receptor and progesterone receptor negative tumours compared to other women (log rank test P-value <0.05). CONCLUSION: Despite an earlier survival advantage for women diagnosed with oestrogen receptor positive tumours, after 5 years of survival, women with oestrogen receptor negative and progesterone receptor negative tumours had better long-term survival outcomes from breast cancer compared to other women. PMID- 15550083 TI - Final report from the ASERNIP-S audit of laparoscopic live-donor nephrectomy. AB - The Australian Safety and Efficacy Register of New Interventional Procedures- Surgical (ASERNIP-S) audit of laparoscopic live-donor nephrectomy commenced in 1999 and concluded in 2003. Six centres in Australia and New Zealand contributed data for 219 donor patients regarding perioperative and postoperative outcome of surgery and short-term follow-up. These data were compared with a recent systematic review of LLDN. The Australasian experience to date compared favourably with worldwide practice, both in terms of efficacy and safety for donors (at least in the short term). The collection of audit data, as this technique was introduced into the Australasian healthcare system, has allowed the local experience with this technique to be pooled and shared, resulting in the development of Australasian practice in line with world's best practice in this area. PMID- 15550084 TI - Choledochal cyst in adults: aetiological considerations to intrahepatic involvement. AB - BACKGROUND: Choledochal cysts are rare anomalies of the biliary tree and their presentation in adults is infrequent. The high incidence of associated anomalous pancreaticobiliary duct junction (APBDJ) has been well documented. However, the prevalence of APBDJ in different anatomic distribution of cysts has yet to be clarified. METHODS: A total 39 adult patients with choledochal cysts were consecutively enrolled: 22 patients had Todani type I cysts, 16 patients had type IVa cysts, and one patient had a type IVb cyst. RESULTS: No statistical differences were found in age, sex or manifestations. A higher frequency of associated APBDJ was identified in patients with type IVa cyst (100%) than those with type I cyst (P < 0.05). With a mean follow-up of 7.2 years after cyst excision, three patients with type I choledochal cyst and four with type IVa cyst have had episodes of recurrent cholangitis, all accompanied with preoperatively diagnosed APBDJ. CONCLUSION: Anomalous pancreaticobiliary duct junction has a close relationship with adult choledochal cysts, particularly type IVa. We propose the hypothesis that intrahepatic involvement results from progressive enzymatic destruction. Adult choledochal cysts in association with APBDJ, with or without intrahepatic component, should be carefully monitored. PMID- 15550085 TI - Efficiency of obliteration procedures in the surgical treatment of hydatid cyst of the liver. AB - BACKGROUND: Hydatid cyst of the liver (HCL) is a parasitic infestation caused by several species of Echinococcus and is endemic in many sheep-raising areas. The aim of the present study is to evaluate the value of clinical, physical and laboratory findings and to discuss the diagnostic and therapeutic options in 250 patients with HCL. METHODS: Between January 1980 and December 1989, 148 patients with HCL were treated surgically, and surgical procedures performed were evaluated retrospectively. On the basis of these findings, a prospective study of 102 patients was initiated between January 1990 and December 1999. RESULTS: External drainage was performed in retrospective and prospective groups, respectively, in 110 and five patients, obliteration procedures in 31 and 82 and resectional procedures in seven and 15. While mean hospitalization time morbidity, mortality and recurrence rates were 16.2 +/- 6.9 days, 37.8%, 2.0% and 6.7% in the retrospective group, these rates were decreased to 7.8 +/- 3.5 days, 10.7%, 0.9% and 1.9%, respectively, in the prospective group. Overall morbidity, mortality and recurrence rates and median hospitalization time were 49.5%, 2.6%, 9.5% and 18.0 +/- 7.4 days in external drainage group, 7.0%, 0% 0.8% and 8.5 +/- 4.5 in obliteration procedures and 9.0%, 4.5%, 0% and 7.3 +/- 1.9 resectional procedures, respectively. CONCLUSIONS: External drainage should be performed only in infected HCL. Resection procedures are too radical and extensive for benign lesions. Obliteration procedures are simple and safe methods for the treatment of HCL, and they have low morbidity, mortality and recurrence rates. PMID- 15550086 TI - Total versus subtotal thyroidectomy for the management of benign multinodular goiter in an endemic region. AB - BACKGROUND: Because controversy still continuous to surround use of total thyroidectomy for the management of benign multinodular goiter, the present study aims to prospectively compare the safety and efficacy of total thyroidectomy with subtotal thyroidectomy. METHODS: A total of 200 consecutive patients with benign multinodular goiter were assigned to have either total thyroidectomy (n = 105) or subtotal thyroidectomy (n = 95) based on preoperative evaluation, intraoperative macroscopic findings and nodular dissemination. The patients with no healthy tissue or nodules localized in the dorsal part of the gland, which are usually left during normal subtotal resection, were assigned to the total thyroidectomy group. Demographic details, biochemical findings, indications for operation, operating time, specimen weight, complications and hospital stay were noted. RESULTS: There was no significant difference in the sex, hormonal status or duration of goiter between the two groups (P = 0.74, P = 0.59 and P = 0.59, respectively). The mean operating time was longer (148.52 min +/- 51.10 vs 135.10 min +/- 32.47, P = 0.03), and the mean weight of the specimens was greater (228.40 g +/- 229.91 vs 157.01 g +/- 151.23, P = 0.01) for total rather than subtotal thyroidectomy. Either temporary recurrent laryngeal nerve (RLN) palsy or hypoparathyroidism occurred in 10 (9.3%) or 12 (11.4%) of the patients undergoing total compared with six (6.3%) or nine (9.5%) of the patients undergoing subtotal thyroidectomy (P = 0.40 and P = 0.65, respectively). Either permanent RLN palsy or hypoparathyroidism was observed in one patient undergoing total thyroidectomy (P = 0.34 for each comparison). The mean hospital stay was longer in the total thyroidectomy group (2.24 days +/- 1.18 vs 1.89 days +/- 0.72 for subtotal thyroidectomy, P = 0.01). CONCLUSIONS: The present study shows that total thyroidectomy can be performed without increasing risk of complication, and it is an acceptable alternative for benign multinodular goiter, especially in endemic regions, where patients present with a huge multinodular goiter. PMID- 15550087 TI - Feasibility of laparoscopic common bile duct exploration in a rural centre. AB - BACKGROUND: Laparoscopic common bile duct exploration has emerged as a preferred option for the management of choledocholithiasis. The present study sought to review the feasibility of this technique in a rural centre. METHODS: A comprehensive retrospective review was undertaken of all patients who underwent surgical treatment of biliary calculi in Lismore, NSW (Australia), between January 1996 and December 2002. RESULTS: During the study period, 1567 consecutive patients underwent laparoscopic cholecystectomy, of whom 82 (5.2%) had choledocholithiasis identified at intraoperative cholangiography. A total of 86 laparoscopic common bile duct explorations were undertaken in these patients, 37 (43%) via a transcystic approach, and 49 (57%) via a laparoscopic choledochotomy. All common bile duct calculi were successfully removed in 78 cases, representing an overall duct clearance rate of 90.7%. Complications were noted in seven patients, a morbidity rate of 8.5%. Median operative time for the procedure over the study period was 173 min. Median hospital stay was 6 days for all patients. CONCLUSIONS: Laparoscopic common bile duct exploration can be successfully undertaken in a rural setting by general surgeons who have appropriate laparoscopic experience, and should be the procedure of choice for the management of choledocholithiasis in these patients. It should not be restricted to specialized surgical departments in major referral centres. PMID- 15550088 TI - Cardiac surgery in octogenarians. AB - BACKGROUND: Early and late results were studied in order to improve the indications for surgery in the elderly. METHODS: Two hundred and thirty-seven patients aged 80 years or older underwent cardiac surgery between 1987 and 2001. The mean age of patients, which included 148 men and 89 women, was 82 years. Elective operations were performed in 194 patients and urgent or emergency operations in 43. Coronary artery bypass grafting (CABG) was performed in 104 patients, valve surgery in 60, CABG plus valve in 58, and other surgery in 15. Late results were obtained in 91% of patients, and the mean follow-up period was 54 months. RESULTS: Operative mortality was 9% in total; 7% in CABG, 5% in valve, 10% in CABG plus valve. Operative mortality was significantly higher in the urgent/emergency group than in the elective group (25% vs 6%). The actuarial survival rate for hospital survivors at 60 months after surgery was 75% and the mean survival period 76 months. There were no significant differences among operations. Preoperatively 81% of the patients had been in New York Heart Association class III or IV, and 88% of survivors were in class I or II in the late period. CONCLUSIONS: Early and late results for elective surgery in octogenarians are satisfactory. However, for urgent or emergent cases, there is a marked increase in morbidity and mortality. PMID- 15550089 TI - Surgical aspects of Operation Bali Assist: initial wound surgery on the tarmac and in flight. AB - BACKGROUND: The explosion of three bombs on 12 October 2002 in Kuta, Bali resulted in mass casualties akin to those seen in war. The aim of the present report is to describe the sequence of events of Operation Bali Assist including triage, resuscitation and initial wound surgery in Bali at Sanglah Hospital in the aeromedical staging facility (ASF), Denpasar airport and the evacuation to Darwin. METHODS: A descriptive report is provided of the event and includes; resuscitation, anaesthesia, initial burns surgery management including escharotomy and fasciotomy, head injury management and importance of supplies and medical records with a description of the evacuation to Darwin. RESULTS: Operation Bali Assist involved five C130 Hercules aircraft and aeromedical evacuation medical and nursing teams managing 66 casualties in the Denpasar area and their evacuation to Royal Darwin Hospital with ketamine the most useful anaesthetic agent and cling film the most useful burns dressing. Twelve procedures were performed at the ASF including seven escharotomies, three fasciotomies and two closed reductions. One escharotomy was performed in flight. DISCUSSION: The important lessons learnt from the exercise is the inclusion of a surgeon in the aeromedical evacuation team, the importance of debridement and delayed primary closure, the usefulness of cling film as a burns dressing and the importance of continuous assessment. Future disaster planning exercises need to consider a patient age mix that might be expected in a shopping mall, rather than the young adult encountered in Bali, a more familiar age mix for Australian Defence Force medical staff. PMID- 15550090 TI - Paediatric trauma teams in Australia. AB - BACKGROUND: Trauma teams have been associated with improved survival probability of paediatric trauma patients. The present study seeks to estimate the use of trauma teams in Australian paediatric tertiary referral centres and describe their medical composition, leadership and criteria for activation. METHODS: Australian paediatric tertiary referral centres were identified. A structured questionnaire assessing the presence, composition and means of activation of a trauma team was mailed to the 'Director, Emergency Department' of all identified hospitals. Three months later, all hospitals were contacted by telephone to complete and verify data collection. RESULTS: Questionnaires were distributed to eight hospitals. Seventy-five per cent had an established trauma team. Hospitals without a trauma team claimed to have insufficient doctors to form a team and insufficient trauma caseload to justify a team. All trauma teams were potentially activated by prehospital paramedic data (field triage) and required a combination of anatomical, physiological and mechanistic criteria for activation. The two methods of mobilizing a trauma team were by dispatching a common call onto individual pagers (66%) or a specific trauma pager (33%) carried by trauma team members. Fifty per cent of hospitals had a two-tier, stratified trauma team response. All teams consisted of emergency, surgical and intensive care unit registrars. Trauma team leaders were emergency medicine specialists/registrars (33%), surgical registrars (33%) and non-defined (33%). Consultant surgeons were not members of any trauma team. Eighty-three per cent of trauma teams consisted of more junior members after hours. Fifty per cent of hospitals did not have a surgical registrar on site outside of business hours. Eighty-eight per cent of hospitals engaged in some form of trauma audit. CONCLUSIONS: Trauma teams are utilized by most Australian paediatric tertiary referral centres, with fairly uniform medical composition and criteria for activation. Paediatric surgeons presently have limited leadership roles and membership of Australian paediatric trauma teams. PMID- 15550091 TI - Peritoneal mesothelial cells produce inflammatory related cytokines. AB - BACKGROUND: Peritonitis involves cascading interactions between cytokines that initiate robust signalling processes via the interferon-g and nuclear factor kappa B pathways. The present study evaluates the interplay between various putative inducers of peritonitis and a battery of inflammation-related cytokines. METHODS: Cultures of peritoneal mesothelial cells were isolated from omenta harvested from male Wistar rats. These cultures were exposed to tumour necrosis factor (TNF)-alpha, lipopolysaccharide, zymosan, myeloperoxidase, peritoneal fluid from rats with zymosan-induced peritonitis, and peritoneal fluid from control animals. The production of TNF-alpha, interleukin (IL)-1beta, IL-6, and IL-10 was assessed after 4, 12 and 24 h. RESULTS: Lipopolysaccharide and zymosan stimulated TNF-alpha, IL-1beta, and IL-10 production; and peritoneal fluids from both control animals and animals with zymosan-induced peritonitis stimulated the production of TNF-alpha, IL-1RII, and IL-6. Expression and secretion of TNF-alpha occurred in a constitutive manner and was regulation at the protein level. The decoy molecule IL-1 receptor type II (IL-1RII) was produced at the same time as IL-1beta and production of the anti-inflammatory cytokine IL-10 was evident within 4 h. IL-6 was constitutively expressed and regulated at the transcriptional level as indicated by a marked discontinuity between the amount of IL-6 produced and the extent IL-6 messenger ribonucleic acid (mRNA) expression. CONCLUSIONS: Tumour necrosis factor-alpha might not be the sole primary mediator of peritonitis. The anti-inflammatory molecules IL-1RII and IL 10 are induced at the same time as the pro-inflammatory cytokines TNF-alpha, IL 1beta, and IL-6. This suggests that complex control systems are set in place by the factors that stimulate peritoneal mesothelial cells and might have the potential to cause peritonitis. PMID- 15550092 TI - Clinicians' reported use of clinical priority assessment criteria and their attitudes to prioritization for elective surgery: a cross-sectional survey. AB - OBJECTIVES: To explore the attitudes of clinicians working in New Zealand publicly funded hospitals towards prioritizing patients for elective surgery, and their reported use of clinical priority assessment criteria (CPAC). DESIGN: A cross-sectional study using a postal questionnaire. The questionnaire drew on themes identified from an earlier qualitative study. Questions were closed and information was sought about perceptions of the need to prioritize patients, effective ways of doing so and the use of CPAC. SETTING: New Zealand. PARTICIPANTS: A national sample of cardiologists, cardiac, general and orthopaedic surgeons, and registrars. RESULTS: Three hundred and thirty-two clinicians responded to the survey (74.1%). Respondents generally agreed that a nationally consistent method of prioritizing patients for surgery was required but felt their clinical judgement was the most effective way of prioritizing patients. Current CPAC were considered to be administrative tools and there was marked variation in their reported use. Consistent use of CPAC using the constructs provided was more likely to be reported by cardiac specialists than general or orthopaedic surgeons. Other features of the hospital system in which surgeons worked also had a major impact on access to elective surgery. CONCLUSIONS: Clinicians recognized the need for a nationally consistent method of prioritizing patients. Although most did not consider current CPAC were effective in achieving this, many felt there was some potential in further development of tools. However, further development is problematic in the absence of objective measures of need and ability to benefit. PMID- 15550093 TI - Adhesional small bowel obstruction after colorectal surgery. AB - BACKGROUND: Acute clinical indicators of complications in colorectal surgery can be readily attained but it is much harder to gather indicators of long-term outcomes such as small bowel obstruction (SBO). However, with improved in hospital data collection and coding such information is becoming available. The aim of the present study was to examine our own hospital database for SBO rates post-colorectal surgery. METHODS: The database was searched and then the relevant medical records were reviewed of all patients admitted to Flinders Medical Centre (Bedford Park, Australia) between July 1999 and November 2002 with a diagnosis of SBO following a colorectal procedure during this same time period. RESULTS: There were 21 patients that accounted for 28 readmissions from a total colorectal procedure group of 583 patients. The arbitrary subgroups were: 13/325 (4%) for colonic resections; 7/186 (3.7%) for rectal resections; 0/12 (0%) for laparotomies/other procedures; and 1/39 (2.6%) for stoma formation/reversals. The overall SBO rate requiring readmission was therefore 3.6%/pt in the 3 years. There was a large variation in the first readmission interval, 38% occurred within 3 months, 43% between 3 and 12 months, and 19% after 1 year. At the first readmission 38% of patients had operative treatment. The mean length of stay was 6.12 days for non-operative vs 21.62 days for operative treatment. CONCLUSION: The reported rate of SBO of 3.6% (at 3 years time interval) is in accordance with other studies. With 38% of patients being treated operatively at first admission there is good acceptance for conservative management in non-strangulated SBO. The prolonged hospital stay for patients needing surgery warrants further investigation. PMID- 15550094 TI - Endoscopic retrieval of a teaspoon from the stomach. PMID- 15550095 TI - Use of Foley's catheter to gain access for retroperitoneoscopy. AB - With the increasing popularity of laparoscopic urologic surgery, many different methods have been used to dissect the extraperitoneal space and gain access to the kidney and ureter. We present our initial experience using a Foley catheter to gain retroperitoneal access. This technique was successfully used in 30 children. We have not encountered any major complications and recommend its use in children. PMID- 15550096 TI - RE: Training of breast surgeons. PMID- 15550097 TI - Validity of reported melanoma incidence rates in New Zealand. PMID- 15550099 TI - Trauma can be an attractive career option. PMID- 15550100 TI - Management of contrast extravasation injury of the forearm during computed tomography. PMID- 15550101 TI - RE: Melanoma lost in the freckle. PMID- 15550102 TI - RE: Being CLEAR: evaluation of a module to teach critical appraisal to surgeons. PMID- 15550103 TI - Delayed-type hypersensitivity to subcutaneous heparin with tolerance of i.v. administration. PMID- 15550104 TI - Primary hepatic carcinosarcoma. PMID- 15550105 TI - Acute anterior compartment syndrome following low energy non-contact injury. PMID- 15550106 TI - Enterobius vermicularis and clinical appendicitis: worms in the vermiform appendix. PMID- 15550107 TI - Mucosa-associated lymphoid tissue lymphoma of the thyroid in a woman with chronic lymphocytic thyroiditis. PMID- 15550108 TI - Soft tissue solitary adult myofibroma in an intervertebral foramen. PMID- 15550109 TI - Splenic infarct: a rare cause of spontaneous rupture leading to massive haemoperitoneum. AB - A 76-year-old woman presented to the accident and emergency department with an acute surgical abdomen. She did not have any significant medical history or history of trauma. Examination findings revealed generalized peritonitis. Preliminary investigations were unhelpful in revealing a cause. On urgent exploratory laparotomy, a ruptured spleen was found and splenectomy performed. The hollow viscera were normal. The gross and microscopic appearances were suggestive of splenic rupture occurring through areas of infarction. We proceed to analyse the causes of spontaneous rupture of the spleen in our discussion of this case. PMID- 15550110 TI - Pre duodenal portal vein as a cause of duodenal obstruction in an adult. AB - Pre duodenal portal vein (PDPV) has not been reported as a cause of duodenal obstruction in adults. We describe a 60 year old man who presented with symptoms of gastric outlet obstruction caused by PDPV, and was diagnosed at laparotomy and treated by gastrojejunostomy. PMID- 15550113 TI - On the molecular mechanism of somatic hypermutation of rearranged immunoglobulin genes. AB - Somatic hypermutation (SHM) diversifies the genes that encode immunoglobulin variable regions in antigen-activated germinal centre B lymphocytes. Available evidence strongly suggests that DNA deamination potentiates phase I SHM and subsequently triggers phase II SHM. A concise review of this evidence is followed by a detailed critique of two possible models which suggest that polymerase-eta potentiates phase II SHM via either its DNA-dependent or its RNA-dependent DNA synthetic activity. Quantitative analysis, in the context of extant data that define the features of SHM, favours the RNA-dependent mechanism. PMID- 15550114 TI - Effect of retinoic acid in experimental diabetic nephropathy. AB - Although the pathogenetic mechanism of diabetic nephropathy has not been elucidated, an inflammatory mechanism has been suggested to contribute to its progression. Monocyte chemoattractant peptide (MCP)-1 attracts macrophages and T cells, and ultimately injures renal tissue. In early diabetic nephropathy, urinary excretion of MCP-1 was elevated, and increased as renal damage became more severe. Podocytes are expected to have an inflammatory role in diabetic nephropathy, as the surface expression of chemokine receptors such as CCR and CXCR on these cells has been recently reported. Although retinoid (retinal), a known anti-inflammatory agent, has been reported to be beneficial in some experimental models of renal disease, it has not been determined to prevent disease progression in diabetic nephropathy. We investigated the effects of all trans retinoic acid on the production of MCP-1 under high glucose conditions in cultured mouse podocytes. We also evaluated whether all-trans retinoic acid inhibits inflammatory changes and improves renal function during the early stages of diabetic nephropathy in streptozotocin-induced diabetic rats. In cultured podocytes, high glucose stimuli rapidly upregulated the MCP-1 mRNA transcript and protein release. Treatment with retinoic acid tended to suppress the MCP-1 gene transcript, and significantly inhibited MCP-1 protein synthesis induced by high glucose stimulation. Urinary protein excretion and the urinary albumin : creatinine ratio (ACR) were significantly higher in diabetic rats 4 weeks after the induction of diabetes mellitus compared with control rats, and retinoic acid treatment markedly decreased both proteinuria and urinary ACR (proteinuria: 1.25+/-0.69 vs 0.78+/-0.72 mg/mgCr, P=0.056; urinary ACR: 0.47+/-0.25 vs 0.21+/ 0.06 mg/mgCr, P=0.088). Urinary excretion of MCP-1 was rapidly increased 2 days after induction of diabetes mellitus in diabetic rats, and further increased until rats were 4 weeks of age, compared with control rats. Retinoic acid treatment resulted in 30% reduction of the urinary level of MCP-1 compared with vehicle-treated diabetic rats (119.3+/-74.2 vs 78.1+/-62.7 pg/mgCr, P=0.078). Immunohistochemistry revealed a significant increase in staining for MCP-1 and anti-monocyte/macrophage (ED-1) protein in the diabetic kidney, and retinoic acid treatment significantly suppressed intrarenal MCP-1 and ED-1 protein synthesis. In conclusion, podocytes are involved in the inflammatory reaction under diabetic circumstances, and these reactions were suppressed by retinoic acid. Retinoic acid also suppressed inflammatory changes in the diabetic rat kidney, and decreased proteinuria in diabetic rats. These results suggest that retinoic acid may have renoprotective effects in the early stages of diabetic nephropathy through an anti-inflammatory activity. PMID- 15550115 TI - Human growth hormone presented by K14hGH-transgenic skin grafts induces a strong immune response but no graft rejection. AB - Although immune responses leading to rejection of transplantable tumours have been well studied, requirements for epithelial tumour rejection are unclear. Here, we use human growth hormone (hGH) expressed in epithelial cells (skin keratinocytes) as a model neo-self antigen to investigate the consequences of antigen presentation from epithelial cells. Mice transgenic for hGH driven from the keratin 14 promoter express hGH in skin keratinocytes. This hGH-transgenic skin is not rejected by syngeneic non-transgenic recipients, although an antibody response to hGH develops in grafted animals. Systemic immunization of graft recipients with hGH peptides, or local administration of stimulatory anti-CD40 antibody, induces temporary macroscopic graft inflammation, and an obvious dermal infiltrate of inflammatory cells, but not graft rejection. These results suggest that a neo-self antigen expressed in somatic cells in skin can induce an immune response that can be enhanced further by induction of specific immunity systemically or non-specific immunity locally. However, immune responses do not always lead to rejection, despite induction of local inflammatory changes. Therefore, in vitro immune responses and in vivo delayed type hypersensitivity are not surrogate markers for immune responses effective against epithelial cells expressing neoantigens. PMID- 15550116 TI - Role for NKG2-A and NKG2-C surface receptors in chronic CD4+ T-cell responses. AB - The participation of CD94 and NKG2 gene family members in the function of NK cells and CD8+ cytolytic cells has recently been addressed in detail. However, the role that these molecules play in the key CD4+ regulatory cells remains largely unexplored. This study has examined the expression and regulation of CD94 and NKG2 genes in purified human peripheral CD4+ cells stimulated with several agents. We found a constitutive expression of NKG2-E in CD94-depleted resting peripheral CD4+ cells, whereas inductions of NKG2-A and NKG2-C required chronic cell activation and occurred after expression of CD94. We found that CD3-mediated stimulation induces the expression of CD94 first by day 5 of culture, followed by NKG2-A by day 15 and finally NKG2-C, which is not detected until 20 days after repeated stimulation. This pattern of gene expression differs sharply from that observed in purified CD8+ T cells, where mRNA from all NKG2 gene family members are detected after 5 days of stimulation. Selective activation of TCR V beta 2 bearing cells with toxic shock syndrome toxin-1 superantigen reveals that mRNA induction of NKG2-A and NKG2-C genes is significantly influenced by the presence of cytokines (IL-10 and TGF-beta) and by the restimulation of the cells. In addition, the occupancy of the CD94/NKG2-A receptor expressed on these superantigen-stimulated CD4+ T lymphocytes abrogates TNF-alpha and IFN-gamma production, whereas NKG2-C enhances production of these cytokines. Taken together our results reveal strict gene regulatory mechanisms for CD94 and NKG2 gene expression on CD4+ cells that are different from those governing the expression of these same genes in CD8+ cells. The results suggest that these genes also participate in chronic CD4+ T-cell responses. PMID- 15550117 TI - Shaping of adaptive immune responses to soluble proteins by TLR agonists: a role for IFN-alpha/beta. AB - Toll-like receptors (TLR) are believed to play a major role in the recognition of invading organisms, although their ability to shape immune responses is not completely understood. Our aim was to investigate in vivo the effect of different TLR stimuli on the generation of antibody responses and the induction of CD8+ T cell cross-priming after immunization with soluble protein antigens. While all TLR agonists tested elicited the production of immunomodulatory cytokines, marked differences were observed in their ability to stimulate antigen-specific immune responses. Zymosan, poly(I:C) and CpG DNA, which signal through TLR2/6, 3 and 9, respectively, were found to strongly induce the production of IgG2a antibodies, whereas R-848 (TLR7) and LPS (TLR4) did so much more weakly. In contrast, LPS, poly(I:C) and CpG DNA, but not zymosan, induced functional CD8+ T-cell responses against OVA; peptidoglycan (TLR2/?) and R-848 were also ineffective in stimulating cross-priming. Experiments using IFN-alpha/beta R-deficient mice showed that the induction of cross-priming by LPS and poly(I:C) was abrogated in the absence of IFN-alpha/beta signalling, and induction by CpG DNA was greatly reduced. Overall, our results identify LPS as another TLR agonist that is able to generate functional cross-priming against a soluble protein antigen. In addition, our results demonstrate that the ability of TLR stimuli to initiate CD8+ T-cell responses against soluble protein antigens is largely dependent on the IFN alpha/beta signalling pathway. PMID- 15550118 TI - Novel adjuvant based on a proteoliposome-derived cochleate structure containing native lipopolysaccharide as a pathogen-associated molecular pattern. AB - Proteoliposomes (PL) from Neisseria meningitidis B have been widely used as a core antigen for antimeningococcal vaccination. PL contain major outer membrane proteins, LPS and phospholipids, and they induce a strong Th1 immune response, but they have low stability in solution. Attending to the need for new vaccine adjuvants, we developed a highly stable cochleate structure (CS) from PL using a technology that allows easy incorporation of new antigens. We explored the ability of PLCS to activate the immune system and its possible application as an adjuvant for parenteral and mucosal routes. Our results showed that PLCS were able to upregulate the expression of MHC class II and costimulatory molecules on human dendritic cells, as well as being able to stimulate the production of soluble mediators of a Th1 response, such as IL-12 and nitric oxide. High levels of anti-PL IgG were detected in serum after i.m. or mucosal (oral and nasal) administration, but also anti-PL secretory IgA was produced in saliva following nasal delivery. The immune response polarization to a Th1 pattern was confirmed by the induction of IgG2a antibodies, positive delayed type hypersensitivity reactions, and IFN-gamma production by splenocytes from immunized mice. The adjuvant potential was explored using PLCS containing ovalbumin (Ova). PLCS-Ova was able to elicit a substantial increase in anti-Ova IgG compared with Ova alone. In addition, a significant reduction in lesion size was observed in mice immunized with Leishmania major antigens in PLCS after challenge with virulent protozoa, suggesting at least partial modulation of the Th2 environment induced by this parasite. In conclusion, our results support the use of PLCS as a potent Th1 adjuvant for parenteral and mucosal vaccines. PMID- 15550119 TI - Inulin-derived adjuvants efficiently promote both Th1 and Th2 immune responses. AB - There has been a recent resurgence of interest into new and improved vaccine adjuvants. This interest has been stimulated by the need for new vaccines to combat problematic pathogens such as SARS and HIV, and to counter potential bioterrorist attacks. A major bottleneck in vaccine development is the low immunogenicity of purified subunit or recombinant proteins, creating the need for safe human adjuvants with high potency. A major problem in the search for the ideal adjuvant is that adjuvants that promote cell-mediated (Th1) immunity (e.g. Freund's complete adjuvant) generally have unacceptable local or systemic toxicity that precludes their use in human vaccines. There is a need for a safe, non-toxic adjuvant that is able to stimulate both cell-mediated and humoral immunity. Inulin-derived adjuvants that principally stimulate the innate immune system through their ability to activate the alternative complement pathway have proven ability to induce both cellular and humoral immunity. With their excellent tolerability, long shelf-life, low cost and easy manufacture, they offer great potential for use in a broad range of prophylactic and therapeutic vaccines. Based on successful animal studies in a broad range of species, human trials are about to get underway to validate the use of inulin-based adjuvants in prophylactic vaccines against hepatitis B, malaria and other pathogens. If such trials are successful, then it is possible that inulin-derived adjuvants will one day replace alum as the adjuvant of choice in most human prophylactic vaccines. PMID- 15550120 TI - Mucosal adjuvants and delivery systems for protein-, DNA- and RNA-based vaccines. AB - Almost all vaccinations today are delivered through parenteral routes. Mucosal vaccination offers several benefits over parenteral routes of vaccination, including ease of administration, the possibility of self-administration, elimination of the chance of injection with infected needles, and induction of mucosal as well as systemic immunity. However, mucosal vaccines have to overcome several formidable barriers in the form of significant dilution and dispersion; competition with a myriad of various live replicating bacteria, viruses, inert food and dust particles; enzymatic degradation; and low pH before reaching the target immune cells. It has long been known that vaccination through mucosal membranes requires potent adjuvants to enhance immunogenicity, as well as delivery systems to decrease the rate of dilution and degradation and to target the vaccine to the site of immune function. This review is a summary of current approaches to mucosal vaccination, and it primarily focuses on adjuvants as immunopotentiators and vaccine delivery systems for mucosal vaccines based on protein, DNA or RNA. In this context, we define adjuvants as protein or oligonucleotides with immunopotentiating properties co-administered with pathogen derived antigens, and vaccine delivery systems as chemical formulations that are more inert and have less immunomodulatory effects than adjuvants, and that protect and deliver the vaccine through the site of administration. Although vaccines can be quite diverse in their composition, including inactivated virus, virus-like particles and inactivated bacteria (which are inert), protein-like vaccines, and non-replicating viral vectors such as poxvirus and adenovirus (which can serve as DNA delivery systems), this review will focus primarily on recombinant protein antigens, plasmid DNA, and alphavirus-based replicon RNA vaccines and delivery systems. This review is not an exhaustive list of all available protein, DNA and RNA vaccines, with related adjuvants and delivery systems, but rather is an attempt to highlight many of the currently available approaches in immunopotentiation of mucosal vaccines. PMID- 15550121 TI - Adjuvant synergy: the effects of nasal coadministration of adjuvants. AB - Modern peptide and protein subunit vaccines suffer from poor immunogenicity and require the use of adjuvants. However, none of the currently licensed adjuvants can elicit cell-mediated immunity or are suitable for mucosal immunization. In this study we explored the immunological effect of nasal co-administration of adjuvants with distinct functions: cholera toxin subunit B, a potent mucosal adjuvant that induces strong humoral responses, muramy di-peptide (MDP), an adjuvant known to elicit cell mediated immunity but rarely used nasally, and chitosan, an adjuvant that achieves specific physiological effects on mucosal membranes that improve antigen uptake. Groups of five female BALB/c mice received on days 1 and 56 nasal instillations of the recombinant Helicobacter pylori antigen urease admixed to single or multiple adjuvant combinations. Serum IgG kinetics were followed over 24 weeks. At the conclusion of the experiment, local antibody responses were determined and antigen-specific recall responses in splenocyte cultures were assayed for proliferation and cytokine production. The combination of adjuvants was shown to further contribute to the increased antigenicity of recombinant H. pylori urease. The data presented here outline and support facilitation of increased immunomodulation by an adjuvant previously defined as an effective mucosal adjuvant (chitosan) for another adjuvant (MDP) that is not normally effective via this route. PMID- 15550122 TI - Cytokines as adjuvants for avian vaccines. AB - The worldwide trend towards a reduced reliance on in-feed antibiotics has increased the pressure to develop alternative strategies to manage infectious diseases in poultry. With this in mind, there is a great emphasis on vaccine use and the enhancement of existing vaccines to provide long-term protection. Currently existing adjuvants for poultry can have deleterious side-effects, such as inflammation, resulting in the down-grading of meat quality and a subsequent reduction in profits. Therefore, to enhance the use of vaccination, alternative adjuvants must be developed. The use of recombinant cytokines as adjuvants in poultry is attracting considerable attention, and their potential role as such has been addressed by several studies. The recent identification of a number of chicken cytokine genes has provided the possibility to study their effectiveness in enhancing the immune response during infection and vaccination. This review focuses on the recent studies involving the assessment of cytokines as vaccine adjuvants. PMID- 15550123 TI - Challenges facing adjuvants for cancer immunotherapy. AB - An adjuvant is defined as a product that increases or modulates the immune response against an antigen (Ag). Based on this general definition many authors have postulated that the ideal adjuvant should increase the potency of the immune response, while being non-toxic and safe. Although dozens of different adjuvants have been shown to be effective in preclinical and clinical studies, only aluminium-based salts (Alum) and squalene-oil-water emulsion (MF59) have been approved for human use. However, for the development of therapeutic vaccines to treat cancer patients, the prerequisites for an ideal cancer adjuvant differ from conventional adjuvants for many reasons. First, the patients that will receive the vaccines are immuno-compromised because of, for example, impaired mechanisms of antigen presentation, non-responsiveness of activated T cells and enhanced inhibition of self-reactivity by regulatory T cells. Second, the tumour Ag are usually self-derived and are, therefore, poorly immunogenic. Third, tumours develop escape mechanisms to avoid the immune system, such as tumour editing, low or non-expression of MHC class I molecules or secretion of suppressive cytokines. Thus, adjuvants for cancer vaccines need to be more potent than for prophylactic vaccines and consequently may be more toxic and may even induce autoimmune reactions. In summary, the ideal cancer adjuvant should rescue and increase the immune response against tumours in immuno-compromised patients, with acceptable profiles of toxicity and safety. The present review discusses the role of cancer adjuvants at the different phases of the generation of antitumour immunity following vaccination. PMID- 15550127 TI - Significance of the cytokine interferon gamma in clinical dermatology. AB - Interferon (IFN)-gamma is considered a key cytokine of innate and adaptive immunity playing pivotal roles in host defence against microbial pathogens and tumours, and exerts profound antiproliferative and antifibrotic effects. In this review we discuss applications and perspectives of IFN-gamma in clinical dermatology, such as papillomavirus and bacterial infections, tumours, atopic dermatitis, and fibrotic conditions such as scleroderma and postradiation fibrosis. Moreover, we give a summary of the pharmacologic properties including main side effects and potential risk factors of IFN-gamma therapy. Although former enthusiasm for IFN-gamma (e.g. in atopic dermatitis) has subsided, this cytokine might remain a promising tool (and target) in clinical dermatology, due to its central immunobiologic functions, better characterization of its kinetics in diseases facilitating optimized treatment schedules, and successful applications in fibrotic conditions such as scleroderma, idiopathic pulmonary and skin postradiation fibrosis. PMID- 15550128 TI - Topical tacrolimus for treatment of childhood vitiligo in Asians. AB - Childhood vitiligo is a common disorder of pigmentation in India. Considering the lack of uniformly effective and safe treatment modalities for children with vitiligo, search for newer therapeutic agents continues. This study was designed to evaluate the role of topical tacrolimus in the treatment of childhood vitiligo. Twenty-five children with vitiligo were treated with topical 0.03% tacrolimus ointment applied twice daily for 12 weeks. Response was noted as marked to complete (> 75% repigmentation), moderate (50-75% repigmentation) and mild (< 50% repigmentation). Twenty-two children (9 boys and 13 girls) of mean age 7.2 +/- 1.4 years completed the study. Twelve (54.5%) children had vitiligo vulgaris, nine (40.9%) had focal vitiligo and one (4.5%) had segmental vitiligo. The mean duration of disease was 8 +/- 3 months. Nineteen (86.4%) children showed some repigmentation at the end of 3 months and other three had no response. Of these 19 children, repigmentation was marked to complete in 11 (57.9%), moderate in five (26.3%) and mild in three (15.7%) children. Side effects were minimal, such as the pruritus and burning noted in only three patients. Topical tacrolimus is an effective and well-tolerated treatment modality in Asian children with vitiligo. PMID- 15550129 TI - Self-detected cutaneous melanomas in Italian patients. AB - Self-detection of suspicious pigmented skin lesion combined with rapid referral to dermatologic centres is the key strategy in the fight against melanoma. The investigation of factors associated with pattern of detection of melanoma (self- vs. nonself-detection) may be useful to refine educational strategies for the future. We investigated the frequency of melanoma self-detection in a Mediterranean population at intermediate melanoma risk. A multicentric survey identified 816 consecutive cases of cutaneous melanoma in the period January to December 2001 in 11 Italian clinical centres belonging to the Italian Multidisciplinary Group on Melanoma. All patients filled a standardized questionnaire and were clinically examined by expert dermatologists. Self detected melanomas were 40.6%, while the remaining lesions were detected by a dermatologist (18.5%), the family physician (15.2%), other specialists (5%), the spouse (12.5%), a friend or someone else (8.2%). Variables associated with self detected melanomas were female sex, young age, absence of atypical nevi, knowledge of the ABCD rule, habit of performing skin self-examination. Self detected melanomas did not differ from nonself-detected tumours in term of lesion thickness; however, patients with self-detected melanomas waited a longer period before having a diagnostic confirmation (patient's delay) (> 3 months: odds ratio, 3.89; 95% confidence interval, 2.74-5.53). In order to reduce the patients' delays, educational messages should adequately stress the need for a prompt referral to a physician once a suspicious pigmented lesion is self detected. PMID- 15550130 TI - The association of smoking and acne in men in Hong Kong and in India: a retrospective case-control study in primary care settings. AB - Previous studies on the association of smoking with acne vulgaris have reported conflicting results. The objective of this study was to investigate such an association. Our setting was three primary care practices in Hong Kong and one primary care practice in India. Patient characteristics in all four practices are similar. All medical records in these practices incorporate a section in which the smoking habits of each patient is routinely documented. We searched our database and retrieved 632 records of patients with acne seen in the previous 5 years. We also retrieved 632 records of age- and sex-matched controls. Fifty three out of 379 male patients with acne and 25 out of 379 male controls were smokers (P = 0.001; OR, 2.3; 95% CI, 1.4-3.8). Six out of 253 female patients with acne and three out of 253 female controls were smokers (OR, 2.0; 95% CI, 0.5 9.4). We conclude that smoking is likely to bear a positive correlation with acne for men. Our numbers are too small for a definite conclusion to be drawn for females. PMID- 15550131 TI - Clinicopathological features and prognostic factors of cutaneous melanoma among Hong Kong Chinese. AB - Cutaneous melanoma is rare among Chinese people. The clinicopathologic features and prognostic factors of 63 Hong Kong ethnic Chinese cases were analysed. We found that the mean age of onset was 65.5 years with a female : male ratio of 1 : 1.33. Forty-two (66.7%) patients had their tumours on their feet. More than 50% had the acral lentiginous histologic subtype. The mean tumour thickness was 5.73 mm with 92.1% (n = 58) having a Clark's level of III or more. Of the tumours, 57.1% were ulcerated. Stage I and II tumours comprised 73% (n = 46) while stage III and IV tumours made up of 26% (n = 17). The 1-, 3- and 5-year survival rates were 75%, 30% and 17%, respectively. Univariate analysis for overall survival showed that advanced clinical staging (stage III and IV), tumour ulceration and thicker tumour (>4 mm) had a significantly poorer prognosis. Multivariate analysis demonstrated that advanced clinical staging was the most decisive prognostic factor followed by tumour ulceration. Our study showed that cutaneous melanoma in Chinese people is a disease predominantly of an older age group with the acral lentiginous histologic type located mainly on the feet. The prognosis depends on clinical staging and ulceration status. PMID- 15550132 TI - Oral squamous cell carcinoma during long-term treatment with hydroxyurea. AB - Hydroxyurea (HU) is commonly used for the treatment of chronic myelogenous leukaemia, polycythemia vera and essential thrombocythaemia. Patients receiving HU present a number of side-effects including skin/mucosa changes and tumours. Mucocutaneous abnormalities include xerosis, ichthyosiform lesions, dark brown pigmentation of skin folds and nails, malleolar ulcers, oral mucositis and oral ulcers. Cutaneous squamous/basal cell carcinomas have also often been reported following long-term administration of HU. HU-induced carcinogenesis is due to both the mutagenic potential of this agent and to an impairment of DNA repair mechanisms after damage by external factors such as ultraviolet radiation. Oral cancer following long-term treatment with HU has been reported only once, in a patient with concomitant multiple skin tumours. We present the unique case of a patient with polycythemia vera who developed oral cancer after 15 years of HU therapy. PMID- 15550133 TI - Parallel fluctuation of anti-desmoglein 3 and anti-BP180 autoantibody titres in a patient with bullous pemphigoid. AB - We report a 73-year-old Japanese female who developed IgG autoantibodies against BP180 as well as desmoglein 3 (Dsg3). She showed tense blisters on the extremities without apparent mucosal involvement and a skin biopsy indicated subepidermal blisters with eosinophilic spongiosis. Her clinical and histologic features indicated the diagnosis of bullous pemphigoid while anti-Dsg3 IgG might not show an apparent pathogenic effect. Interestingly, titres of anti-Dsg3 IgG fluctuated in parallel with those of anti-BP180 IgG throughout the course with two flares. Although the exact mechanism for autoantibody production is still unknown, the close link in the production of IgG autoantibodies against two independent skin antigens suggests a shared immunoregulatory mechanism against cutaneous autoantigens. PMID- 15550135 TI - Clear cell sarcoma associated with multiple pulmonary metastases with structural and numerical karyotypic changes. AB - Cytogenetic analysis was performed on a specimen from a pulmonary metastasis of clear cell sarcoma originated on the right ankle of a 53-year-old Japanese woman. It revealed near-triploid with several numerical changes including abnormalities in the copy number of chromosomes 7, 8, and 22, and structural abnormalities of chromosome 22 and others. This is a minor case which showed the multiple abnormalities of chromosomes in the absence of a t (12;22) translocation. PMID- 15550134 TI - Fatal toxic epidermal necrolysis due to lansoprazole. AB - Toxic epidermal necrolysis (TEN) is a serious cutaneous reaction and is most commonly drug induced. It is associated with significant morbidity and mortality. We describe a patient who developed fatal TEN after re-exposure to lansoprazole. Three years previously he presented with erythema multiforme due to the same drug. To our knowledge this is the first published report of TEN occurring with lansoprazole. PMID- 15550136 TI - Glaucoma induced by periorbital topical steroid use--a rare complication. AB - Eye complications arising during systemic and ophthalmic steroid use are well known. In contrast, there is little highlighting the similar risks associated with topical steroid use around the eyelids. We now describe a 29-year-old lady who used topical steroids in prolonged fashion throughout most of her life for severe eczema, with the recent application of large quantities of potent steroid continuously for one month, including to the periorbital region. Soon after, she presented to her ophthalmologist with severe bilateral glaucoma and irreversible visual loss attributed to the steroid use. Here we emphasise the possible risks of periorbital topical steroid use, as well as the importance of patient education and ophthalmological follow-up. PMID- 15550137 TI - Nail infestation by Liposcelis bostrychophila Badonnel. AB - We report an unusual case of nail infestation by Liposcelis bostrychophila Badonnel in a 70-year-old woman with onychomycosis. Liposcelis spp., also known as booklouse, are tiny insects that feed on fungi, lichen and decaying materials. In this case, the loosened hyperkeratotic nail provided a favourable environment for these insects. This is the second report of human infestation by Liposcelis spp. PMID- 15550138 TI - Hidradenitis suppurativa, Dowling-Degos and multiple epidermal cysts: a new follicular occlusion triad. AB - This case illustrates the rare association between hidradenitis suppurativa (HS) and Dowling-Degos disease (DDD). Furthermore the association of HS, DDD and multiple epidermal cysts has not to our knowledge been described before, but their coexistence in the same patient is likely to reflect the same follicular anomaly. It is possible that a single underlying defect of follicular proliferation may account for the coexistence of these conditions. PMID- 15550139 TI - Lues maligna. AB - The incidence of syphilis in the UK is rising, particularly in HIV-positive men. We describe a Caucasian, HIV-negative woman who presented with secondary syphilis taking the form known as lues maligna. She also demonstrated the prozone phenomenon whereby the rapid plasmin reagin test was negative, but the Venereal Disease Research Laboratory and Treponema pallidum haemagglutination tests were positive. PMID- 15550140 TI - Elevated MIA levels in the serum of pregnant women and of children. AB - Recent studies have shown that the protein MIA (melanoma inhibitory activity) is a potent serum marker for malignant melanoma. MIA is expressed in chondrocytes at high levels, and might potentially be elevated during periods of growth in childhood. Therefore, we evaluated MIA serum levels in pregnant women and in growing children. We found that no significant enhancement of MIA serum levels was apparent until 38 weeks of pregnancy. Starting at 38 weeks, a significant increase was noted. The study further revealed that children and teenagers also have increased MIA serum levels. However, from the age of 17 years MIA serum levels are not different from those in healthy adults. In summary, MIA can be used as a serum marker for melanoma in adults starting at the age of 17 years. In pregnant women (> 38th week), children, and teenagers MIA should not be used in serum diagnostics for malignant melanoma until larger studies have been performed to generate cut-off levels for each group. PMID- 15550141 TI - Localized granulomatous reaction to a semi-permanent hyaluronic acid and acrylic hydrogel cosmetic filler. AB - Dermalive, an injectable skin filler composed of a combination of synthetic hyaluronic acid and acrylic hydrogel particles was recently developed for soft tissue augmentation. Dermalive produces longer term results than temporary injectable fillers and is associated with a reportedly low incidence of adverse reactions. We describe a marked local reaction to the injection of Dermalive in the nasolabial fold developing within 4 months with histological confirmation of a granulomatous response. To our knowledge there has been only one previous report of a local granulomatous reaction to Dermalive. PMID- 15550143 TI - Brachioradial pruritus--an enigmatic entity. AB - Brachioradial pruritus (BRP) is an unusual clinical entity resulting in pruritus of the upper arms, classically affecting the skin overlying the proximal heads of the brachioradialis muscles. Seven patients with BRP are described. The upper limb was involved in six patients, being bilateral in four and restricted to the right arm in two. Radiological evidence of cervical vertebral osteoarthritis was seen in five. Three patients reported exacerbation with ultraviolet radiation (UVR), i.e., sunlight. All seven were treated with topical capsaicin and four got significant relief. Two of the remaining three patients found relief from amitryptiline. UVR may act as a trigger in some patients although underlying neurological cervical injury also seems to be an important aetiological factor. PMID- 15550142 TI - Vancomycin-induced linear IgA bullous disease presenting as toxic epidermal necrolysis. AB - Linear IgA bullous dermatosis (LABD) is a rare autoimmune vesiculobullous disorder characterized by variable clinical presentations that may mimic bullous pemphigoid, dermatitis herpetiformis, cicatricial pemphigoid and erythema multiforme. A few cases of drug-induced LABD that clinically resembled toxic epidermal necrolysis (TEN) have been reported. A subset of patients with LABD have been found to be drug-induced; the most common drug being vancomycin. The diagnosis of LABD is confirmed by the presence of a linear band of IgA along the basement membrane zone on direct immunofluorescence microscopy. We report a case of a 77-year-old man who presented to us with vancomycin-induced LABD that presented clinically as TEN. He had a complete recovery over a 3-week period following discontinuation of the vancomycin and the addition of oral dapsone therapy. It is important to be aware that drug-induced LABD can mimic TEN. PMID- 15550144 TI - Imiquimod induced regression of clinically diagnosed superficial basal cell carcinoma is associated with early infiltration by CD4 T cells and dendritic cells. AB - Imiquimod is presumed to clear basal cell carcinoma (BCC) through apoptosis mediated by cytokines and lymphocytes, with erosion often observed correlating with complete clearance. The objective was to determine the cellular immune response early in the course of treatment in order to examine whether cell mediated immunity could be responsible for imiquimod mediated regression of BCC. Sixteen adults with clinically diagnosed BCC were openly assigned to 5 days per week of drug (1, 2 or 4 weeks) or placebo (2 weeks) in groups of four. No baseline biopsy was performed. Post-treatment excision specimens were examined by routine and immunohistochemical staining. Treatment was associated with the early appearance of CD4 cells, activated dendritic cells and macrophages, with later infiltration by CD8 T cells. Dendritic cells continually increased with time, while macrophages reached a maximum at 1 week and then declined slightly. There were comparatively few neutrophils or gammadelta T cells. Early infiltrates were most prominent in the tumour and upper dermis. The results are consistent with a cell mediated immune response being responsible for the clearance of the BCC. Several immune-mediated tumour destruction mechanisms are likely to be involved. PMID- 15550145 TI - NK-1 antagonist CP99994 inhibits stress-induced mast cell degranulation in rats. AB - Mast cells are implicated in stress-induced inflammatory skin diseases such as psoriasis. Mechanisms of stress-induced mast cell degranulation however, are not entirely clear. Here we explore the role of activation of a Substance P (SP) receptor (NK-1) on mast cell degranulation upon exposure to stress in rats. A specific nonpeptide NK-1 antagonist, CP99994 was used to treat the rats either peripherally or intracerebroventricularly. Because increased SP activity in the brain may mediate the stress response, we also examined cutaneous mast cell degranulation after central injection of SP. Stress, as well as SP injected centrally, increased mast cell degranulation. Both central and peripheral injection of CP99994 prevented stress-induced mast cell degranulation. Surprisingly, the combination of stress with SP decreased mast cell degranulation, suggesting that high levels of SP may counteract the stress responses. Results in this animal model suggest that NK-1 antagonists may be used therapeutically to treat stress-induced inflammatory skin diseases; however, drug doses should be chosen carefully. PMID- 15550146 TI - Normal head-hair length is correlated with its diameter. AB - Shed head-hair fibres of young (16-20-year-old), nonalopecic women (n = 25), exhibiting both exogen clubs and anagen tips (EA) were studied. Such fibres are shown, for the first time, to comprise approximately 44% of shed hair and to form a uni-modal, positively skewed distribution with a mean length of 16.7 +/- 4.9 cm, which is also correlated with the length of the haircut. As individual fibres exit the skin in early anagen VI, their major-axis diameters increase rapidly to maxima at about 25% of their total potential length and subsequently decrease to their exogen clubs, at a rate of 1.31% per cm (n = 28). EA diameters are further correlated with their lengths. Maximal and proximal diameters increase by 1.40% per cm and 1.02% per cm increments in fibre lengths, respectively (P < 0.0001 each; n = 14), these changes being also different from each other (P < 0.001). Besides identifying and characterizing a new class of normal hair (EA) which will probably feature prominently in future hair research, this study reveals several other important aspects of hair growth: (i) the classically described concept of hair miniaturization in androgenetic alopecia (AGA) is excessively broad and should therefore be revised; (ii) female AGA need not necessarily require a mechanism for rapid miniaturization as recently proposed; and (iii) the putative large variability of normal hair diameters is significantly overestimated, which further opens the field of hair diameter evaluation as a biological indicator of disease and physiological function. PMID- 15550147 TI - Homing receptor and chemokine receptor on intraepidermal T cells in psoriasis vulgaris. AB - Intraepidermal T lymphocytes found in psoriatic skin lesions are involved in the development and maintenance of lesional pathology. It has become clear that differential expression of homing and chemokine receptors determines the specific migration of T cells to distinct tissues and microenvironments, including psoriasis lesions. The aim of the present study was to clarify expression of homing (CLA, VLA-4, and LFA-1) and chemokine (CCR4, CCR6, CCR7, and CXCR3) receptors on intraepidermal T cells in psoriatic lesions using flow cytometry. The vast majority of intraepidermal T cells in psoriatic lesions expressed CLA and LFA-1, whereas 58% of CD4+ and 85% of CD8+ T cells expressed VLA-4. The majority of CD4+ T cells and about half of the CD8+ T cells expressed CCR4 and CCR6, whereas less than one-third of CD4+ and CD8+ T cells expressed CXCR3 or CCR7. In patients with psoriasis the percentages of T cells expressing CLA, CCR4, and CCR6 were much higher in the epidermis of psoriatic plaques than in the peripheral blood. Thus, CLA, CCR4, and CCR6 may play a more important role in the migration of T cells to psoriatic epidermis. PMID- 15550148 TI - Altered expression of L-arginine metabolism pathway genes in chronic wounds in recessive dystrophic epidermolysis bullosa. AB - Individuals with the severe, mutilating Hallopeau-Siemens form of recessive dystrophic epidermolysis bullosa (HS-RDEB) have trauma-induced blisters and skin erosions which often progress to wounds that are slow to heal. These chronic wounds cause considerable morbidity and there is an increased risk of squamous cell carcinoma arising in the wound margins. Currently, little is known about the keratinocyte cell biology in these wounds. Therefore, we compared the gene expression profiles of wound edge with nonwounded skin from two individuals with HS-RDEB. Trauma-induced wound sites had been present in both patients for more than 3 months. Hybridizations using DermArray gene expression filters showed relative differences in gene expression between wounded and unwounded skin. Notably, there was a fivefold increase in expression of arginase-1 (ARG1) in the chronic wound samples. Expression of seven other genes relevant to L-arginine metabolism also showed differences greater than twofold. L-arginine is known to have a critical role in the synthesis of nitric oxide as part of normal tissue repair. Although alterations in arginase isoenzymes have been detected previously in other chronic wounds (human and animal models), this is the first study to demonstrate differences in several components of the L-arginine metabolism pathway in chronic wounds, and the first to examine chronic wounds in HS-RDEB. The data show that the cascade of L-arginine metabolites is altered in HS-RDEB and the findings may provide new insight into the pathology of chronic wounds in this genodermatosis. PMID- 15550149 TI - ADULT ectodermal dysplasia syndrome resulting from the missense mutation R298Q in the p63 gene. AB - Several ectodermal dysplasia syndromes have been shown to result from mutations in the gene that encodes the transcription factor p63. We describe an 11-year-old boy, with clinically normal parents, who had a developmental disorder that resembled EEC (ectrodactyly ectodermal dysplasia-clefting) syndrome (OMIM 604292). He had ectrodactyly and missing middle fingers bilaterally, onychodysplasia, hypodontia with missing teeth, hypohidrosis and lacrimal duct obstruction. DNA sequencing disclosed a heterozygous G-->A substitution at nucleotide 893, that converts an arginine residue (CGA) to glutamine (CAA), the mutation being designated R298Q. This mutation occurs within the DNA-binding domain of p63, and is close to many of the published EEC syndrome mutations. However, R298Q has been described once previously in a large German pedigree, not with EEC syndrome, but another ectodermal dysplasia disorder, ADULT (acro-dermato ungual-lacrimal-tooth) syndrome (OMIM 103285). Further clinical assessment in our patient revealed that, apart from not having cleft lip and/or palate, he had an exfoliative dermatitis of his hands and feet, and some freckling on his face and shoulders. Collectively, these features support a diagnosis of ADULT syndrome. This study has identified a specific genotype-phenotype correlation in a rare ectodermal dysplasia syndrome and the findings are useful in improving genetic counselling in this family. PMID- 15550150 TI - Necrotizing fasciitis due to Serratia marcescens. PMID- 15550151 TI - Topical tacrolimus for oral cicatricial pemphigoid. PMID- 15550152 TI - Dermatoscopic features of benign sebaceous proliferation. PMID- 15550153 TI - Plantar basal cell carcinoma: a possible eccrine origin. PMID- 15550155 TI - Systemic reaction associated with Iramo scyphopolyp, Stephanoscyphus racemosum Komai. PMID- 15550154 TI - Toxic epidermal necrolysis treated with low-dose intravenous immunoglobulin: immunohistochemical study of Fas and Fas-ligand expression. PMID- 15550156 TI - Mallory-Weiss syndrome secondary to vomiting caused by wheat-induced anaphylaxis. PMID- 15550157 TI - Progressive pigmented purpura (Schamberg's disease) responding to TL01 ultraviolet B therapy. PMID- 15550158 TI - Lichen sclerosus of the lip and buccal mucosa. PMID- 15550159 TI - Attitudes of men with androgenetic alopecia to sun protection. PMID- 15550160 TI - Eruptive syringoma in four Black South African children. PMID- 15550162 TI - Dermatologists and cosmetic surgery--a personal view of regulation and training issues. PMID- 15550163 TI - A lesion present at birth. PMID- 15550164 TI - Necrotic ulceration during haemodialysis. PMID- 15550165 TI - A case of wrinkled skin. PMID- 15550166 TI - Evaluation of a communication skills seminar for students in a Japanese medical school: a non-randomized controlled study. AB - BACKGROUND: Little data exist for the effectiveness of communication skills teaching for medical students in non-English speaking countries. We conducted a non-randomized controlled study to examine if a short intensive seminar for Japanese medical students had any impact on communication skills with patients. METHODS: Throughout the academic year 2001-2002, a total of 105 fifth-year students (18 groups of 5 to 7 students) participated, one group at a time, in a two-day, small group seminar on medical interviewing. Half way through the year, a five-station objective structured clinical examination (OSCE) was conducted for all fifth-year students. We videotaped all the students' interaction with a standardized patient in one OSCE station that was focused on communication skills. Two independent observers rated the videotapes of 50 students who had attended the seminar and 47 who had not. Sixteen core communication skills were measured. Disagreements between raters were resolved by a third observer's rating. RESULTS: There was a statistically significant difference in proportions of students who were judged as 'acceptable' in one particular skill related to understanding patient's perspectives: asking how the illness or problems affected the patient's life, (53% in the experimental group and 30% in the control group, p = .02). No differences were observed in the other 15 core communication skills, although there was a trend for improvement in the skill for asking the patient's ideas about the illness or problems (60% vs. 40%, p = .054) and one of the relationship building skills; being attentive and empathic nonverbally (87% vs. 72%, p = .064). CONCLUSION: The results of this study suggest that a short, intensive small group seminar for Japanese medical students may have had a short term impact on specific communication skills, pertaining to understanding patient's perspectives. PMID- 15550167 TI - GOtcha: a new method for prediction of protein function assessed by the annotation of seven genomes. AB - BACKGROUND: The function of a novel gene product is typically predicted by transitive assignment of annotation from similar sequences. We describe a novel method, GOtcha, for predicting gene product function by annotation with Gene Ontology (GO) terms. GOtcha predicts GO term associations with term-specific probability (P-score) measures of confidence. Term-specific probabilities are a novel feature of GOtcha and allow the identification of conflicts or uncertainty in annotation. RESULTS: The GOtcha method was applied to the recently sequenced genome for Plasmodium falciparum and six other genomes. GOtcha was compared quantitatively for retrieval of assigned GO terms against direct transitive assignment from the highest scoring annotated BLAST search hit (TOPBLAST). GOtcha exploits information deep into the 'twilight zone' of similarity search matches, making use of much information that is otherwise discarded by more simplistic approaches. At a P-score cutoff of 50%, GOtcha provided 60% better recovery of annotation terms and 20% higher selectivity than annotation with TOPBLAST at an E value cutoff of 10(-4). CONCLUSIONS: The GOtcha method is a useful tool for genome annotators. It has identified both errors and omissions in the original Plasmodium falciparum annotation and is being adopted by many other genome sequencing projects. PMID- 15550169 TI - Use of the bootstrap in analysing cost data from cluster randomised trials: some simulation results. AB - BACKGROUND: This work has investigated under what conditions confidence intervals around the differences in mean costs from a cluster RCT are suitable for estimation using a commonly used cluster-adjusted bootstrap in preference to methods that utilise the Huber-White robust estimator of variance. The bootstrap's main advantage is in dealing with skewed data, which often characterise patient costs. However, it is insufficiently well recognised that one method of adjusting the bootstrap to deal with clustered data is only valid in large samples. In particular, the requirement that the number of clusters randomised should be large would not be satisfied in many cluster RCTs performed to date. METHODS: The performances of confidence intervals for simple differences in mean costs utilising a robust (cluster-adjusted) standard error and from two cluster-adjusted bootstrap procedures were compared in terms of confidence interval coverage in a large number of simulations. Parameters varied included the intracluster correlation coefficient, the sample size and the distributions used to generate the data. RESULTS: The bootstrap's advantage in dealing with skewed data was found to be outweighed by its poor confidence interval coverage when the number of clusters was at the level frequently found in cluster RCTs in practice. Simulations showed that confidence intervals based on robust methods of standard error estimation achieved coverage rates between 93.5% and 94.8% for a 95% nominal level whereas those for the bootstrap ranged between 86.4% and 93.8%. CONCLUSION: In general, 24 clusters per treatment arm is probably the minimum number for which one would even begin to consider the bootstrap in preference to traditional robust methods, for the parameter combinations investigated here. At least this number of clusters and extremely skewed data would be necessary for the bootstrap to be considered in favour of the robust method. There is a need for further investigation of more complex bootstrap procedures if economic data from cluster RCTs are to be analysed appropriately. PMID- 15550168 TI - Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq. AB - BACKGROUND: Isopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl diphosphate (GGPP) synthase is an important branch point enzyme in terpenoid biosynthesis. Therefore, GGPP synthase is thought to be a key enzyme in biosynthesis of forskolin. Herein we report the first confirmation of the GGPP synthase gene in Coleus forskohlii Briq. RESULTS: The open reading frame for full-length GGPP synthase encodes a protein of 359 amino acids, in which 1,077 nucleotides long with calculated molecular mass of 39.3 kDa. Alignments of C. forskohlii GGPP synthase amino acid sequences revealed high homologies with other plant GGPP synthases. Several highly conserved regions, including two aspartate-rich motifs were identified. Transient expression of the N-terminal region of C. forskohlii GGPP synthase-GFP fusion protein in tobacco cells demonstrated subcellular localization in the chloroplast. Carotenoid production was observed in Escherichia coli harboring pACCAR25DeltacrtE from Erwinia uredovora and plasmid carrying C. forskohlii GGPP synthase. These results suggested that cDNA encoded functional GGPP synthase. Furthermore, C. forskohlii GGPP synthase expression was strong in leaves, decreased in stems and very little expression was observed in roots. CONCLUSION: This investigation proposed that forskolin was synthesised via a non-mevalonate pathway. GGPP synthase is thought to be involved in the biosynthesis of forskolin, which is primarily synthesised in the leaves and subsequently accumulates in the stems and roots. PMID- 15550170 TI - Validation of a prefractionation method followed by two-dimensional electrophoresis - Applied to cerebrospinal fluid proteins from frontotemporal dementia patients. AB - BACKGROUND: The aim of this study was firstly, to improve and validate a cerebrospinal fluid (CSF) prefractionation method followed by two-dimensional electrophoresis (2-DE) and secondly, using this strategy to investigate differences between the CSF proteome of frontotemporal dementia (FTD) patients and controls. From each subject three ml of CSF was prefractionated using liquid phase isoelectric focusing prior to 2-DE. RESULTS: With respect to protein recovery and purification potential, ethanol precipitation of the prefractionated CSF sample was found superior, after testing several sample preparation methods.The reproducibility of prefractionated CSF analyzed on 2-D gels was comparable to direct 2-DE analysis of CSF. The protein spots on the prefractionated 2-D gels had an increased intensity, indicating a higher protein concentration, compared to direct 2-D gels. Prefractionated 2-DE analysis of FTD and control CSF showed that 26 protein spots were changed at least two fold. Using mass spectrometry, 13 of these protein spots were identified, including retinol-binding protein, Zn-alpha-2-glycoprotein, proapolipoproteinA1, beta-2 microglobulin, transthyretin, albumin and alloalbumin. CONCLUSION: The results suggest that the prefractionated 2-DE method can be useful for enrichment of CSF proteins and may provide a new tool to investigate the pathology of neurodegenerative diseases. This study confirmed reduced levels of retinol binding protein and revealed some new biomarker candidates for FTD. PMID- 15550171 TI - Using GIS to establish a public library consumer health collection. AB - BACKGROUND: Learning the exact demographic characteristics of a neighborhood in which a public library serves, assists the collection development librarian in building an appropriate collection. Gathering that demographic information can be a lengthy process, and then formatting the information for the neighborhood in question becomes arduous.As society ages and the methods for health care evolve, people may take charge of their own health. With this prospectus, public libraries should consider creating a consumer health collection to assist the public in their health care needs. Using neighborhood demographic information can inform the collection development librarians as to the dominant age groups, sex, and races within the neighborhood. With this information, appropriate consumer health materials may be assembled in the public library. METHODS: In order to visualize the demographics of a neighborhood, the computer program ArcView GIS (geographic information systems) was used to create maps for specified areas. The neighborhood data was taken from the U.S. Census Department's annual census and library addresses were accumulated through a free database. After downloading the census block information from http://www.census.gov/geo/www/tiger/ the data was manipulated with ArcView GIS and queried to produce maps displaying the requested neighborhood demographics to view in respect to libraries. RESULTS: ArcView GIS produced maps displaying public libraries and requested demographics. After viewing the maps the collection development librarian can see exactly what populations are served by the library and adjust the library's collection accordingly. CONCLUSIONS: ArcView GIS can be used to produce maps displaying the communities that libraries serve, spot boundaries, be it "man-made or natural," that exist prohibiting customer service, and assist collection development librarians in justifying their purchases for a dedicated consumer health collection or resources in general. PMID- 15550172 TI - The Israeli strain IS-98-ST1 of West Nile virus as viral model for West Nile encephalitis in the Old World. AB - West Nile virus (WNV) recently became a major public health concern in North America, the Middle East, and Europe. In contrast with the investigations of the North-American isolates, the neurovirulence properties of Middle-Eastern strains of WNV have not been extensively characterized. Israeli WNV strain IS-98-ST1 that has been isolated from a white stork in 1998, was found to be highly neuroinvasive in adult C57BL/6 mice. Strain IS-98-ST1 infects primary neuronal cells from mouse cortex, causing neuronal death. These results demonstrate that Israeli strain IS-98-ST1 provides a suitable viral model for WNV-induced disease associated with recent WNV outbreaks in the Old World. PMID- 15550173 TI - Merkel cell carcinoma in a malignant pleural effusion: case report. AB - BACKGROUND: Merkel cell (neuroendocrine) carcinoma is a small round blue cell malignant neoplasm that primarily presents in the skin. The diagnosis of Merkel cell carcinoma in a pleural fluid is challenging because of the morphological similarity to many other malignant neoplasms. Immunohistochemical stains can be essential to establish the diagnosis of Merkel cell carcinoma. CASE PRESENTATION: A 77 year-old woman presented with a mass in her right buttock thought clinically to be a boil or sebaceous cyst. Upon histopathologic review including immunohistochemical analysis, a diagnosis of Merkel cell carcinoma was rendered. Wide-excision and sentinel lymph node biopsy revealed negative margins and no evidence of metastasis. Ten months later she complained of bone pain and a bone scan revealed multiple lesions. An abdominal CT scan revealed a T4 vertebral mass and local radiotherapy was administered. Two months later the patient presented with shortness of breath. A chest radiograph showed an effusion and thoracentesis was performed. The fluid was confirmed to contain metastatic Merkel cell carcinoma by cytology and immunohistochemical analysis. CONCLUSIONS: Merkel cell carcinoma is an aggressive neoplasm that can, despite careful surgical management, occasionally present as a malignant pleural effusion in a relatively short time period. Immunohistochemical analysis can aid in confirming this rare outcome. PMID- 15550174 TI - The docking protein Gab1 is the primary mediator of EGF-stimulated activation of the PI-3K/Akt cell survival pathway. AB - BACKGROUND: Gab1 is a docking protein that recruits phosphatidylinositol-3 kinase (PI-3 kinase) and other effector proteins in response to the activation of many receptor tyrosine kinases (RTKs). As the autophosphorylation sites on EGF receptor (EGFR) do not include canonical PI-3 kinase binding sites, it is thought that EGF stimulation of PI-3 kinase and its downstream effector Akt is mediated by an indirect mechanism. RESULTS: We used fibroblasts isolated from Gab1-/- mouse embryos to explore the mechanism of EGF stimulation of the PI-3 kinase/Akt anti-apoptotic cell signaling pathway. We demonstrate that Gab1 is essential for EGF stimulation of PI-3 kinase and Akt in these cells and that these responses are mediated by complex formation between p85, the regulatory subunit of PI-3 kinase, and three canonical tyrosine phosphorylation sites on Gab1. Furthermore, complex formation between Gab1 and the protein tyrosine phosphatase Shp2 negatively regulates Gab1 mediated PI-3 kinase and Akt activation following EGF receptor stimulation. We also demonstrate that tyrosine phosphorylation of ErbB3 may lead to recruitment and activation of PI-3 kinase and Akt in Gab1-/- MEFs. CONCLUSIONS: The primary mechanism of EGF-induced stimulation of the PI-3 kinase/Akt anti-apoptotic pathway occurs via the docking protein Gab1. However, in cells expressing ErbB3, EGF and neuroregulin can stimulate PI-3 kinase and Akt activation in a Gab1-dependent or Gab1-independent manner. PMID- 15550175 TI - Are both sympatric species Ilex perado and Ilex canariensis secretly hybridizing? Indication from nuclear markers collected in Tenerife. AB - BACKGROUND: Intra-specific and intra-individual polymorphism is frequently observed in nuclear markers of Ilex (Aquifoliaceae) and discrepancy between plastid and nuclear phylogenies is the rule in this genus. These observations suggest that inter-specific plastid or/and nuclear introgression played an important role in the process of evolution of Ilex. With the aim of a precise understanding of the evolution of this genus, two distantly related sympatric species collected in Tenerife (Canary Islands), I. perado and I. canariensis, were studied in detail. Introgression between these two species was previously never reported. One plastid marker (the atpB-rbcL spacer) and two nuclear markers, the ribosomal internal transcribed spacer (ITS) and the nuclear encoded plastid glutamine synthetase (nepGS) were analyzed for 13 and 27 individuals of I. perado and I. canariensis, respectively. RESULTS: The plastid marker is intra specifically constant and correlated with species identity. On the other hand, whereas the nuclear markers are conserved in I. perado, they are highly polymorphic in I. canariensis. The presence of pseudogenes and recombination in ITS sequences of I. canariensis explain this polymorphism. Ancestral sequence polymorphism with incomplete lineage sorting, or past or recent hybridization with an unknown species could explain this polymorphism, not resolved by concerted evolution. However, as already reported for many other plants, past or recent introgression of an alien genotype seem the most probable explanation for such a tremendous polymorphism. CONCLUSIONS: Data do not allow the determination with certitude of the putative species introgressing I. canariensis, but I. perado is suspected. The introgression would be unilateral, with I. perado as the male donor, and the paternal sequences would be rapidly converted in highly divergent and consequently unidentifiable pseudogenes. At least, this study allows the establishment of precautionary measures when nuclear markers are used in phylogenetic studies of genera having experienced introgression such as the genus Ilex. PMID- 15550176 TI - Use of a highly sensitive two-dimensional luminescence imaging system to monitor endogenous bioluminescence in plant leaves. AB - BACKGROUND: All living organisms emit spontaneous low-level bioluminescence, which can be increased in response to stress. Methods for imaging this ultra-weak luminescence have previously been limited by the sensitivity of the detection systems used. RESULTS: We developed a novel configuration of a cooled charge coupled device (CCD) for 2-dimensional imaging of light emission from biological material. In this study, we imaged photon emission from plant leaves. The equipment allowed short integration times for image acquisition, providing high resolution spatial and temporal information on bioluminescence. We were able to carry out time course imaging of both delayed chlorophyll fluorescence from whole leaves, and of low level wound-induced luminescence that we showed to be localised to sites of tissue damage. We found that wound-induced luminescence was chlorophyll-dependent and was enhanced at higher temperatures. CONCLUSIONS: The data gathered on plant bioluminescence illustrate that the equipment described here represents an improvement in 2-dimensional luminescence imaging technology. Using this system, we identify chlorophyll as the origin of wound-induced luminescence from leaves. PMID- 15550178 TI - What is the treatment of choice for Tl squamous cell carcinoma of the larynx? PMID- 15550177 TI - CLOE: identification of putative functional relationships among genes by comparison of expression profiles between two species. AB - BACKGROUND: Public repositories of microarray data contain an incredible amount of information that is potentially relevant to explore functional relationships among genes by meta-analysis of expression profiles. However, the widespread use of this resource by the scientific community is at the moment limited by the limited availability of effective tools of analysis. We here describe CLOE, a simple cDNA microarray data mining strategy based on meta-analysis of datasets from pairs of species. The method consists in ranking EST probes in the datasets of the two species according to the similarity of their expression profiles with that of two EST probes from orthologous genes, and extracting orthologous EST pairs from a given top interval of the ranked lists. The Gene Ontology annotation of the obtained candidate partners is then analyzed for keywords overrepresentation. RESULTS: We demonstrate the capabilities of the approach by testing its predictive power on three proteomically-defined mammalian protein complexes, in comparison with single and multiple species meta-analysis approaches. Our results show that CLOE can find candidate partners for a greater number of genes, if compared to multiple species co-expression analysis, but retains a comparable specificity even when applied to species as close as mouse and human. On the other hand, it is much more specific than single organisms co expression analysis, strongly reducing the number of potential candidate partners for a given gene of interest. CONCLUSIONS: CLOE represents a simple and effective data mining approach that can be easily used for meta-analysis of cDNA microarray experiments characterized by very heterogeneous coverage. Importantly, it produces for genes of interest an average number of high confidence putative partners that is in the range of standard experimental validation techniques. PMID- 15550179 TI - 2',5'-Oligoadenylate synthetase activity analysis and human papilloma virus typing as prognostic factors in patients with recurrent respiratory papillomatosis. AB - OBJECTIVE: Determination of early prognostic factors in patients with recurrent respiratory papillomatosis is extremely important, so the major goal of our prospective, multicentre study was to evaluate (1) the feasibility of various factors to determine prognosis of the clinical course, as well as (2) the response to interferon-alpha therapy in recurrent respiratory papillomatosis. METHODS: Forty-two patients with recurrent respiratory papillomatosis were treated with interferon-alpha (3 MU/m(2) three times per week; mean therapy duration was 2.7 +/- 1.8 years) in 1983-1994 and followed-up until 2003. Human papilloma virus (HPV) type, recurrent respiratory papillomatosis severity and 2',5'-oligoadenylate synthetase activity were determined by standard methods and analysed for correlation with the results of long-term clinical outcome. RESULTS AND CONCLUSION: Patients with HPV type 11, a severity score >4, a high number of surgical procedures prior to interferon-alpha therapy and a high basal 2',5' oligoadenylate synthetase activity should be considered at high risk of an aggressive clinical course, often with spread to lower airway passages, malignant transformation and death. Human papilloma virus type, score for recurrent respiratory papillomatosis severity, number of surgical procedures and 2',5' oligoadenylate synthetase activity showed significant association with response to interferon-alpha therapy and the long-term clinical course, so these factors have value in predicting prognosis in recurrent respiratory papillomatosis. PMID- 15550180 TI - Does occurrence of keratinizing stratified squamous epithelium in the middle-ear cavity always indicate a cholesteatoma? AB - The origin and behaviour of keratinizing stratified squamous epithelium, an essential component of cholesteatoma occurring in the middle-ear cavity, has puzzled otologists for decades. In this experimental study in 16 cats, central (n = 23) and peripheral (n = 9) tympanic membrane perforations were observed for up to 63 days before sacrifice. The tympanic membranes with bony rim were excised, decalcified and embedded in Epon 812. Sections were stained with toluidine blue and examined using a light microscope. The perforation had been sealed by meatal epithelium exhibiting pronounced hyperplasia and keratin formation, lying on a bed of granulation tissue. Subtotal central perforations healed within 14 days, forming a bowl-shaped tympanic membrane and leaving parts of the handle of the malleus (with meatal epithelium) protruding freely into the middle-ear cavity. Stratified squamous epithelium, morphologically identical with that of external ear canal epidermis, could be observed on the malleus even 63 days after operation. This meatal epithelium was non-keratinizing, non-invasive, and showed no destructive properties typical of acquired cholesteatoma. During certain circumstances, the cell cycle of hyperplastic epidermal epithelium within the middle-ear cavity can evidently be arrested and inactivated by a local defence mechanism. PMID- 15550181 TI - Navigation as a quality management tool in cochlear implant surgery. AB - This cadaver study assessed the value of navigation in cochlear implant surgery. Cochlear implantation was simulated on a cadaver using a Stryker-Leibinger navigation system and a Nucleus 24 Contour implant. A conventional surgical strategy consisting of mastoidectomy, posterior tympanotomy, and cochleostomy was performed. The navigated surgical procedure was evaluated for accuracy, reliability, reproducibility, and practicability. The technology of computer assisted surgery is applicable in cochlear implantation and beneficial in as much as the navigation-controlled implantation constitutes a non-invasive instrument of quality management. Nevertheless, in order to keep the point accuracy below one millimeter, a referencing method using concealed bordering anatomical structures may be further needed to perform the cochleostomy reliably under the guidance of a navigation system. More reproducible reference systems are needed if navigated lateral skull base surgery is to be fully relied upon. PMID- 15550182 TI - Cervicofacial necrotizing fasciitis: can we expect a favourable outcome? AB - Necrotizing fasciitis of the head and neck is an uncommon, progressive, destructive soft tissue infection of mixed aerobic and anaerobic organisms, having high mortality if left untreated (22 to 100 per cent). This study makes an attempt to analyse various factors and management methods determining the overall prognosis. A retrospective analysis of all cases of necrotizing fasciitis involving the head and neck, with exclusion of those involving the eyelid and the scalp, was undertaken. Various parameters such as demography, aetiology, complications, management and outcome were studied. Males outnumbered the females with the latter having a greater risk of involvement after 60 years. Odontogenic infection was the primary source of infection. Anaerobes were cultured in seven out of 17 cases, with six others showing mixed Gram positive and Gram negative organisms. Anaemia was the most commonly associated illness, with diabetes affecting four out of 17 cases. Aggressive surgical debridement with triple antibiotic therapy was used in the management of necrotizing fasciitis with an overall mortality of 11.8 per cent. Patients having late referral, anaemia and one or other complication had increased duration of total hospital stay. Better results can be obtained with proper control of infection by early diagnosis, aggressive surgical debridement and triple antibiotic therapy, along with timely control of complications and associated illnesses. PMID- 15550183 TI - The myth of tracheomalacia and difficult intubation in cases of retrosternal goitre. AB - Although it is widely believed that management of the airway is difficult in surgery for retrosternal goitre, a review of the literature, revealing management of 1969 patients with retrosternal goitre, provided scant evidence of difficult intubation or post-operative tracheomalacia resulting in tracheal collapse. This was reflected in our own series of 18 thyroidectomies for retrosternal goitre performed at our hospitals. PMID- 15550184 TI - Botulinum toxin: helpful adjunct to early resolution of laryngeal granulomas. AB - Treating laryngopharyngeal reflux is the most accepted treatment for laryngeal granulomas. However, response to this treatment is prolonged and in some cases persistent. The authors hypothesize that this is due to the continuous trauma to the mucoperichondrium from adduction of the vocal folds thus preventing regeneration of healthy mucosa. A prospective study was performed on six patients with laryngeal granulomas failing prolonged laryngopharyngeal reflux treatment, speech therapy, and surgical excisions. Botulinum toxin A was injected into the affected thyroarytenoid to reduce adduction trauma. Video-stroboscopy was used to assess response. A 50 per cent reduction in the size of the granulomas was noted within two weeks with a complete and permanent response within two to eight weeks in five out of six patients. One patient had an obstructing pyogenic granuloma that required excision and recurred after excision but responded to a repeated injection. The addition of a single injection of botulinum toxin A to the standard treatment expeditiously eliminated persistent and recurrent laryngeal granulomas. PMID- 15550185 TI - Wegener's granulomatosis and subglottic stenosis: management of the airway. AB - Wegener's granulomatosis is a multisystemic disease characterized by foci of necrotizing vasculitis and granuloma formation. Subglottic stenosis may occur either as a presenting feature or a late-stage manifestation of the disease, but will occur in approximately 10-20 per cent of cases. We present a series of seven cases of Wegener's granulomatosis with subglottic stenosis and discuss our management of this condition. Where there is active disease, tracheostomy is the first-line surgical treatment of respiratory obstruction, as an adjunct to full medical therapy. More aggressive or elaborate surgical treatments should be reserved for non-active cases in which patients have not required medical treatment for one year. PMID- 15550186 TI - Multidimensional assessment of voice and speech after supracricoid laryngectomy with cricohyoidopexy. AB - This study was designed: to evaluate the vocal function in the patients with supracricoid laryngectomy (SCL) compared with normal subjects; to determine the factors affecting voice (such as number of arytenoid(s) preserved and movement of larynx and tongue base); and to determine the correlations between videolaryngostroboscopy, acoustic and perceptual parameters. Ten patients who underwent SCL with cricohyoidopexy for primary laryngeal squamous cell carcinoma were included into the study. Vocal function was investigated by means or videolaryngostroboscopy. Voice quality was assessed by means of objective acoustic analysis and subjective perceptual ratings by trained raters. Aberrant, incompetent, and rough mucosal wave was observed in the anterior and superior surfaces of arytenoids(s), the inferior part of tongue base and the lateral walls of the hypopharynx. The acoustic parameters were found to be significantly different from those of normal subjects. The values of perceptual scores were approximately within 50 per cent of normal range. The number of arytenoids spared did not affect acoustic or perceptual measurements. A rough, breathy, unpleasant but intelligible and acceptable voice could be obtained after SCL with cricohyoidopexy. PMID- 15550187 TI - Is a doctor needed in the adult ENT pre-admission clinic? AB - OBJECTIVES: Pre-admission clinics are traditionally run jointly by nurses and doctors. Within an adult ENT pre-admission clinic, we wished to assess what doctors added to nurses' pre-clerking, to determine whether doctors were actually needed in the clinic. METHODS: Prospective study, looking at how often doctors, seeing patients after ward-based nurses, changed or added to clerking or tests as organized by nurses. RESULTS: Out of 184 patients, doctors changed or added to nurses' clerking or planned investigations in 47 patients (26 per cent), making 64 different changes. The commonest reasons for changes were ordering blood tests (22 changes), chest X-rays (eight), cancelling due to hypertension (seven), altering drug history (five) and requesting electrocardiograms (five changes). CONCLUSION: Most changes made by doctors could be eliminated by designing a pre admission clinic protocol that could easily be used by nurses. We recommend that all ENT departments consider implementing nurse-led pre-admission clinics. PMID- 15550188 TI - Concha bullosa: reducing middle meatal adhesions by preserving the lateral mucosa as a posterior pedicle flap. AB - BACKGROUND: Concha bullosa, an extensively pneumatized middle turbinate, may obstruct the paranasal sinuses. Messerklinger's partial lateral turbinectomy is commonly used to debulk the concha bullosa, leaving a raw surface with the potential for adhesions. MATERIALS AND METHODS: A modified technique of partial lateral turbinectomy is described. A posterior pedicled mucosal flap covers the inferior raw surface of the medial lamella of the middle turbinate. Three-month follow up of a consecutive series is compared with concurrent controls. RESULTS: Two (7 per cent) of 28 posterior pedicled flap and four (21 per cent) of 19 traditional partial lateral turbinectomies developed mild middle meatal adhesions (p = 0.011). Posterior pedicled flap reduced the need for post-operative cleaning of the middle meatus. CONCLUSION: The posterior pedicled mucosal flap is a simple modification to partial lateral turbinectomy that covers the raw surface facing the lateral nasal wall, significantly reducing adhesions and speeding recovery. PMID- 15550189 TI - Disaster avoided: otalgia warns of potential electrode extrusion. AB - Cochlear implantation is usually a safe procedure but, as in most surgical interventions, it can carry some risk of complications. These can be divided into major and minor that, in turn, in turn can present early or late. This case report highlights a potential late, major complication of cochlear implantation namely extrusion, which was averted by prompt intervention. The patient presented with a three months' history of otalgia some six years after successful implantation. The posterior half of the tympanic membrane was found to be retracted on the electrode, which appeared to be on the point of extruding through the drum. A tragal cartilage myringoplasty was performed to separate the electrode from the medial surface of the tympanic membrane. There was immediate and lasting relief of the otalgia, the electrode was well protected and performance with the device was unchanged. The authors recommend reinforcing an atrophic tympanic membrane at the time of the cochlear implantation by a cartilage graft to avoid this potentially serious complication. The case also highlights the need for regular otoscopy for all implanted patients. PMID- 15550190 TI - Role of CO2 laser in the management of obstructive ectopic lingual thyroids. AB - Ectopic lingual thyroid glands may occasionally undergo massive hypertrophy and be a source of airway obstruction. Securing the airway prior to surgery in such cases may be difficult. In the present paper, we report on two cases of obstructive ectopic lingual thyroids which were successfully managed endoscopically using CO(2) laser. The use of CO(2) laser for removal of ectopic lingual thyroid tissue offers advantages over traditional surgical techniques. In our opinion, removal of an appropriate amount of gland to secure an adequate airway is all that is required in most cases. PMID- 15550191 TI - Paget's disease and cochlear implantation. AB - Paget's disease of bone is a common disorder of unresolved etiology characterized by excessive bone resorption followed by excessive bone formation. If the skull is affected this may result in hearing loss and eventually develop into profound deafness. To date, no cases of cochlear implantation in patients with Paget's disease have been reported. The authors present a case of radiographically confirmed Paget's disease of the skull in a 77-year-old man with a 20-year history of progressive bilateral sensorineural hearing loss who underwent cochlear implantation. A successful insertion of the Nucleus 24 Contour electrode array was achieved without surgical and postoperative complications. At the 10 months' postoperative evaluation, the patient had gained useful open-set speech perception. In quiet conditions, his performance scores on the word and sentence recognition tests were 100 and 98 per cent, respectively. In the presence of noise (at +10 dB. signal-to-noise ratio), his performance scores on the word and sentence recognition tests were 96 and 94 per cent, respectively. PMID- 15550192 TI - Nd: YAG laser treatment for adult hypopharyngeal haemangioma. AB - Adult hypopharyngeal haemangioma is an uncommon neoplasm. We present three cases of adult hypopharyngeal haemangioma which were successfully treated with a neodymium: yttrium-aluminium-garnet (Nd: YAG) laser in our hospital between 2000 and 2003. The patients in this series experienced no untoward events from the use of the laser. Excessive bleeding and airway problems were not encountered. Postoperative pain was minimal. Excellent long-term function and a full resolution of symptoms can be obtained using Nd: YAG laser. The laser provides a bloodless field and complete healing of the hypopharynx, without stenosis or scar formation. PMID- 15550193 TI - Otoscopic appearances and tympanometric changes in narghile smokers. AB - Narghile (water-pipe) smoking requires the generation of significant negative intrapharyngeal pressure, which may be transmitted to the middle ear through the Eustachian tube. A total of 80 ears from regular narghile smokers were examined otoscopically and by tympanometry. Seventy ears from heavy cigarette smokers were similarly examined and served as a control group. There was a highly significant increase in the prevalence of attic retractions (P < 0.01) in the narghile smokers. The tympanometric changes were not significantly different between the two groups (P > 0.05). PMID- 15550194 TI - Teflon-induced granuloma: a source of false positive positron emission tomography and computerized tomography interpretation. AB - Patients diagnosed with malignancy often undergo combined positron emission tomography (PET) and computerized tomography (CT) to investigate possible metastases. This report presents a case in which, in the investigation of suspected pulmonary malignancy, combined PET and CT images suggested a malignant lesion at the level of the vocal fold. Biopsy of the lesion, however, confirmed the clinical diagnosis of a Teflon granuloma. The case highlights the potential for a false positive report during scanning of patients who have had vocal fold injection. PMID- 15550195 TI - Lower oesophageal meat bolus clearance using a radiologically guided balloon catheter: case in a 94-year-old patient. AB - Lower oesophageal foreign body meat bolus obstruction is potentially life threatening. We report a case in a 94-year-old woman in which conservative measures and flexible oesophagoscopy were unsuccessful. Rigid oesophagoscopy was considered technically difficult and so clearance by interventional radiology was attempted. Through the mouth a radiologically guided balloon catheter was introduced. It was passed beyond the bolus to dilate the site of obstruction, before being withdrawn and inflated above the bolus, pushing it into the stomach. Although successful in this case, the technique is previously unreported and so its complication rate is unknown. It is therefore presented only to be considered when other treatments are neither effective nor possible. PMID- 15550196 TI - Preoperative evaluation of bone destruction using three-dimensional computed tomography in cholesteatoma. AB - This study investigated the usefulness of three-dimensional computed tomography (3DCT) in the evaluation of bony defects caused by cholesteatoma. Fifteen patients with chronic otitis media with cholesteatoma who showed bony destruction or suspicious destruction in two-dimensional CT were examined using 3DCT. The CT data were transferred to a workstation with a real-time image processor. We used three-dimensional reconstruction software enabling image processing. In all patients, 3DCT clearly delineated the destruction of bony structures by cholesteatoma. The 3DCT-generated images provided spatial relationships, which were not easily appreciated on two-dimensional CT. Intraoperative bony destruction findings correlated with 3DCT findings. From these results, 3DCT could be useful to evaluate the invasiveness of cholesteatoma to the cranial base. It could also be helpful in planning reconstruction during surgery. PMID- 15550197 TI - Trachoma decline and widespread use of antimicrobial drugs. AB - Trachoma is disappearing in many parts of the world, even in the absence of specific control programs. Following mass antimicrobial drug treatments for trachoma in western Nepal, the prevalence of trachoma declined far more rapidly than could be attributed to the control program alone. Pharmacy surveys in the same region found that children received more antichlamydial drugs from sources outside the trachoma program than they did from the program itself. We demonstrate that high background antimicrobial drug use may be responsible for much of the observed decline in trachoma and discuss its potential role in eliminating this infectious disease. PMID- 15550198 TI - Public health interventions and SARS spread, 2003. AB - The 2003 outbreak of severe acute respiratory syndrome (SARS) was contained largely through traditional public health interventions, such as finding and isolating case-patients, quarantining close contacts, and enhanced infection control. The independent effectiveness of measures to "increase social distance" and wearing masks in public places requires further evaluation. Limited data exist on the effectiveness of providing health information to travelers. Entry screening of travelers through health declarations or thermal scanning at international borders had little documented effect on detecting SARS cases; exit screening appeared slightly more effective. The value of border screening in deterring travel by ill persons and in building public confidence remains unquantified. Interventions to control global epidemics should be based on expert advice from the World Health Organization and national authorities. In the case of SARS, interventions at a country's borders should not detract from efforts to identify and isolate infected persons within the country, monitor or quarantine their contacts, and strengthen infection control in healthcare settings. PMID- 15550199 TI - Topographic changes in SARS coronavirus-infected cells at late stages of infection. AB - Scanning electron and atomic force microscopy was used for the first time to view the maturation of the severe acute respiratory syndrome-associated coronavirus at the cell surface. The surface form of the cells at advanced infection displayed prolific pseudopodia that, in addition to the rest of the plasma membrane, were also active sites of virus release. High magnification of the maturing virus particles showed a rosette appearance with short knoblike spikes under both the scanning electron and atomic force microscopes. The final expulsion step of the maturing virus particles seemed to result in some disruptions to the plasma membrane. The cytoskeletal network along the edge of the infected cells was enhanced and could be involved in transporting and expelling the progeny virus particles. Thickening of the actin filaments at the cell edge provided the bending force to extrude the virus particles. PMID- 15550200 TI - Evaluating human papillomavirus vaccination programs. AB - Human papillomavirus (HPV) has been implicated as the primary etiologic agent of cervical cancer. Potential vaccines against high-risk HPV types are in clinical trials. We evaluated vaccination programs with a vaccine against HPV-16 and HPV 18. We developed disease transmission models that estimated HPV prevalence and infection rates for the population overall, by age group, by level of sexual activity within each age group, and by sex. Data were based on clinical trials and published and unpublished sources. An HPV-16/18 vaccine for 12-year-old girls would reduce cohort cervical cancer cases by 61.8%, with a cost-effectiveness ratio of 14,583 dollars per quality-adjusted life year (QALY). Including male participants in a vaccine rollout would further reduce cervical cancer cases by 2.2% at an incremental cost-effectiveness ratio of 442,039 dollars/QALY compared to female-only vaccination. Vaccination against HPV-16 and HPV-18 can be cost effective, although including male participants in a vaccination program is generally not cost-effective, compared to female-only vaccination. PMID- 15550201 TI - Enhanced identification of postoperative infections among inpatients. AB - We evaluated antimicrobial exposure, discharge diagnoses, or both to identify surgical site infections (SSI). This retrospective cohort study in 13 hospitals involved weighted, random samples of records from 8,739 coronary artery bypass graft (CABG) procedures, 7,399 cesarean deliveries, and 6,175 breast procedures. We compared routine surveillance to detection through inpatient antimicrobial exposure (> 9 days for CABG, > 2 days for cesareans, and > 6 days for breast procedures), discharge diagnoses, or both. Together, all methods identified SSI after 7.4% of CABG, 5.0% of cesareans, and 2.0% of breast procedures. Antimicrobial exposure had the highest sensitivity, 88%-91%, compared with routine surveillance, 38%-64%. Diagnosis codes improved sensitivity of detection of antimicrobial exposure after cesareans. Record review confirmed SSI after 31% to 38% of procedures that met antimicrobial surveillance criteria. Sufficient antimicrobial exposure days, together with diagnosis codes for cesareans, identified more postoperative SSI than routine surveillance methods. This screening method was efficient, readily standardized, and suitable for most hospitals. PMID- 15550202 TI - Enhanced identification of postoperative infections among outpatients. AB - We investigated using administrative claims data to identify surgical site infections (SSI) after breast surgery and cesarean section. Postoperative diagnosis codes, procedure codes, and pharmacy information were automatically scanned and used to identify claims suggestive of SSI ("indicators") among 426 (22%) of 1,943 breast procedures and 474 (10%) of 4,859 cesarean sections. For 104 breast procedures with indicators explained in available medical records, SSI were confirmed for 37%, and some infection criteria were present for another 27%. Among 204 cesarean sections, SSI were confirmed for 40%, and some criteria were met for 27%. The extrapolated infection rates of 2.8% for breast procedures and 3.1% for cesarean section were similar to those reported by the National Nosocomial Infection Surveillance program but differ in representing predominantly outpatient infections. Claims data may complement other data sources for identification of surgical site infections following breast surgery and cesarean section. PMID- 15550204 TI - Nucleocapsid protein as early diagnostic marker for SARS. AB - Serum samples from 317 patients with patients with severe acute respiratory syndrome (SARS) were tested for the nucleocapsid (N) protein of SARS-associated coronavirus, with sensitivities of 94% and 78% for the first 5 days and 6-10 days after onset, respectively. The specificity was 99.9%. N protein can be used as an early diagnostic maker for SARS. PMID- 15550203 TI - Histopathologic improvement with lymphedema management, Leogane, Haiti. AB - In countries where bancroftian filariasis is endemic, lymphedema of the leg is a public health problem, particularly for women, who are disproportionately affected. We investigated the effect of basic lymphedema management (hygiene, skin care, and lower limb movement and elevation) on the histologic features of lymphedema. A total of 118 skin-punch biopsy specimens were collected from the legs of 91 patients enrolled in a lymphedema treatment clinic in Leogane, Haiti. Follow-up biopsy specimens were collected from 27 patients succeeds, equals 12 months later. Keratinocyte hyperproliferation, condensed dermal collagen, and mononuclear perivascular infiltrate increased with lymphedema stage, which suggested progressive chronic inflammation and fibrosis. Follow-up biopsies showed reductions in perivascular mononuclear infiltrate in the superficial dermis (41% decrease in prevalence), perivascular fibrosis in the deep dermis (58% decrease), and periadnexal mononuclear infiltrate (53% decrease). These data suggest that the clinical improvement commonly observed with basic lymphedema management has a histologic basis. PMID- 15550205 TI - Human infection caused by Clostridium hathewayi. AB - We describe a 27-year-old man with acute cholecystitis, hepatic abscess, and bacteremia caused by Clostridium hathewayi, a newly described gram-negative, endospore-forming, rod-shaped bacterium. This report is the first of human infection caused by this microorganism. PMID- 15550206 TI - Commercial logging and HIV epidemic, rural Equatorial Africa. AB - We found a high seroprevalence of HIV among young women in a commercial logging area in Cameroon. The vulnerability of these young women could be related to commercial logging and the social and economic networks it induces. The environmental changes related to this industry in Equatorial Africa may facilitate HIV dissemination. PMID- 15550207 TI - Burkholderia cenocepacia vaginal infection in patient with smoldering myeloma and chronic hepatitis C. AB - We report a case of a vaginal infection caused by a strain of Burkholderia cenocepacia. The strain was isolated from vaginal swab specimens from a 68-year old woman with smoldering myeloma and chronic hepatitis C virus infection who was hospitalized for abdominal abscess. Treatment with piperacillin/tazobactam eliminated B. cenocepacia infection and vaginal symptoms. PMID- 15550208 TI - Decreased levofloxacin susceptibility in Haemophilus influenzae in children, Hong Kong. AB - Among 563 strains of Haemophilus influenzae from young children in Hong Kong, 5 (0.9%) had decreased susceptibility to quinolones. The five strains had a Ser-84 Lys or Asp-88-Asn substitution in GyrA. Pulsed-field gel electrophoresis showed that the isolates are genetically diverse. PMID- 15550209 TI - Steps for preventing infectious diseases in women. AB - Communicable diseases account for approximately 25% of deaths in most Latin American and Caribbean countries; illness from communicable diseases reaches 40% in developing countries. Mainly affected are poor women in rural areas. A medical approach is not sufficient to implement effective infectious disease prevention strategies in women, which would offset these numbers. Health policies must be changed, and social restrictions that circumscribe women need to be eliminated. In the long run, the only solution is to improve women's socioeconomic status. The following three steps are necessary for developing a prevention strategy: 1) a gender perspective must be incorporated into infectious disease analysis and research to target policies and programs. Data collected must be disaggregated by sex, age, socioeconomic status, education, ethnicity, and geographic location; 2) models must be developed and implemented that address gender inequities in infectious diseases in an integrated manner; and 3) outreach activities must be supported, using information, education, and communication strategies and materials for advocacy and training. Active participation of civil society groups is key to translating the strategy into specific interventions. PMID- 15550210 TI - Barriers to infectious disease care among lesbians. AB - Despite the considerable number of women in the United States who identify as lesbian, few data exist that address lesbians' health needs. The Institute of Medicine emphasized that data on sexually transmitted infections, Pap smear screening, and cervical dysplasia among lesbians were needed to guide clinical practice, policy development, and patient education. Use of surveillance data for this purpose is limited because risk classifications exclude same-gender sex among women or subsume it under behavior considered as higher risk. However, sexual transmission of human papillomavirus, HIV, Treponema pallidum, and Trichomonas vaginalis between women has been reported. Data indicate that lesbians receive routine Pap smear screening less frequently than is optimal. Moreover, lesbians commonly report previous pregnancy, induced abortion, and hormonal contraceptive use. Education of lesbians and their care providers should counter assumptions that sex between women confers no risk for transmission of sexually transmitted infections, and lesbians should receive Pap smears according to current guidelines. PMID- 15550211 TI - Gender and monitoring the response to HIV / AIDS pandemic. AB - The mechanisms, techniques, and data sources used to monitor and evaluate global AIDS prevention and treatment services may vary according to gender. The Joint United Nations Programme on HIV / AIDS has been charged with tracking the response to the pandemic by using a set of indicators developed as part of the Declaration of Commitment endorsed at the U.N. General Assembly Special Session on AIDS in 2001. Statistics on prevalence and incidence indicate that the pandemic has increasingly affected women during the past decade. Women's biologic, cultural, economic, and social status can increase their likelihood of becoming infected with HIV. PMID- 15550212 TI - Nurses' working conditions: implications for infectious disease. AB - Staffing patterns and nurses' working conditions are risk factors for healthcare associated infections as well as occupational injuries and infections. Staffing shortages, especially of nurses, have been identified as one of the major factors expected to constrain hospitals' ability to deal with future outbreaks of emerging infections. These problems are compounded by a global nursing shortage. Understanding and improving nurses' working conditions can potentially decrease the incidence of many infectious diseases. Relevant research is reviewed, and policy options are discussed. PMID- 15550213 TI - Vaccines for women age 50 and older. AB - For older populations, most of whom are women, preventing illnesses and deaths through the use of vaccines is a leading public health challenge. Our understanding about how age and sex affect the immune system is limited, and basic and translational research aimed at improving vaccines and immune responses of older persons is needed. In the meantime, fully implementing current vaccine recommendations, particularly those for influenza and pneumococcal vaccines, can save thousands of lives and prevent illnesses in persons > 50 years of age. PMID- 15550214 TI - Sexual power and HIV risk, South Africa. AB - Gender power inequities are believed to play a key role in the HIV epidemic through their effects on women's power in sexual relationships. We hypothesized that lack of sexual power, measured with a four-point relationship control scale and by a woman's experience of forced sex with her most recent partner, would decrease the likelihood of consistent condom use and increase the risk for HIV infection among sexually experienced, 15- to 24-year-old women in South Africa. While limited sexual power was not directly associated with HIV, it was associated with inconsistent condom use: women with low relationship control were 2.10 times more likely to use condoms inconsistently (95% confidence interval [CI] 1.17-3.78), and women experiencing forced sex were 5.77 times more likely to use condoms inconsistently (95% CI 1.86-17.91). Inconsistent condom use was, in turn, significantly associated with HIV infection (adjusted odds ratio 1.58, 95% CI 1.10-2.27). PMID- 15550215 TI - Women and autoimmune diseases. AB - Autoimmune diseases affect approximately 8% of the population, 78% of whom are women. The reasons for the high prevalence in women are unknown, but circumstantial evidence links autoimmune diseases with preceding infections. Animal models of autoimmune diseases have shown that infections can induce autoimmune disease. For example, coxsackievirus B3 (CB3) infection of susceptible mice results in inflammation of the heart (myocarditis) that resembles myocarditis in humans. The same disease can be induced by injecting mice with heart proteins mixed with adjuvant(s), which indicates that an active infection is not necessary for the development of autoimmune disease. We have found that CB3 triggers autoimmune disease in susceptible mice by stimulating elevated levels of proinflammatory cytokines from mast cells during the innate immune response. Sex hormones may further amplify this hyperimmune response to infection in susceptible persons, which leads to an increased prevalence of autoimmune diseases in women. PMID- 15550216 TI - Contribution of sex-linked biology and gender roles to disparities with trachoma. AB - Globally, trachoma is the leading infectious cause of blindness. Survey data consistently show that trachoma-related blindness is two to four times higher in women than men. Tracing the increased risk for trachoma and its consequences for women suggests that other factors besides biology may contribute. Understanding the reasons for the excess risk for and consequences of trachoma in girls and women requires examining a number of issues: Are girls and women more biologically susceptible to the consequences of infection with Chlamydia trachomatis? Could other factors help explain the excess of conjunctival scarring and trichiasis in women? Do gender roles affect the risk for trachoma and its consequences? Are women more likely to have recurrence after trichiasis surgery compared to men? This article explores the answers to these questions. PMID- 15550217 TI - Rubella elimination and improving health care for women. AB - In the Americas, the Pan American Health Organization supports strategies for the appropriate control and elimination of vaccine-preventable diseases, especially if the strategies are designed to reduce health inequities, strengthen the political commitment for immunization services, and promote the culture of prevention. In September 2003, the 44th Directing Council of the Pan American Health Organization adopted a goal to eliminate rubella and congenital rubella syndrome by 2010. One of the main objectives of this initiative is improving women's health, consistent with achieving the Millennium Development Goals. An important component of rubella elimination is conducting vaccination campaigns for men and women of childbearing age to reduce rapidly the number of people susceptible to rubella infection. From 1998 to 2002, the English-speaking Caribbean, Chile, Costa Rica, Brazil, and Honduras conducted mass rubella vaccination campaigns aimed at adults. Vaccination coverage reached > 95% in each country with an exception of the Caribbean, where the coverage was 80%. PMID- 15550218 TI - Globalization and infectious diseases in women. AB - Women have an enhanced vulnerability to disease, especially if they are poor. Indeed, the health hazards of being female are widely underestimated. Economic and cultural factors can limit women's access to clinics and health workers. The World Health Organization (WHO) reports that less is spent on health care for women and girls worldwide than for men and boys. As a result, women who become mothers and caretakers of children and husbands often do so at the expense of their own health. The numbers tell the story: the latest (2003) World Health Report showed that, globally, the leading causes of death among women are HIV/AIDS, malaria, complications of pregnancy and childbirth, and tuberculosis. PMID- 15550219 TI - Smallpox models as policy tools. AB - Mathematical models can help prepare for and respond to bioterrorism attacks, provided that their strengths and weaknesses are clearly understood. A series of initiatives within the Department of Health and Human Services brought modelers together with biologists and epidemiologists who specialize in smallpox and experts in bioterrorism response and health policy and has led to the parallel development of models with different technical approaches but standardized scenarios, parameter ranges, and outcome measures. Cross-disciplinary interactions throughout the process supported the development of models focused on systematically comparing alternate intervention strategies, determining the most important issues in decision-making, and identifying gaps in current knowledge. PMID- 15550220 TI - [Bioethics and cancer]. PMID- 15550221 TI - [Lymphatic mapping and biopsy of sentinel lymph node in patients with breast cancer. Results of the first phase of a study]. AB - BACKGROUND: The standard surgical treatment in breast cancer patient is resection of the primary tumor and axillary lymphadenectomy; nevertheless almost 50% of patients without axillary palpable nodes do not have axillary metastasis in the axillary dissection specimen. In theory the sentinel node identification (lymphatic mapping) selects patients with high risk of hidden metastasis and avoids unnecessary axillary dissections, the technique is performed with blue dye, radio colloid or both. OBJECTIVE: To know the sentinel node value in axillary staging in patients with invasive breast cancer without palpable nodes. MATERIAL AND METHODS: Prospective study including 87 breast cancer patients stage I-II without palpable nodes. In 65 we use blue dye only and in 22 blue dye and Tc99 nanocoloid, all patients were submitted to axillary dissection; the sentinel node was study by imprint cytology, frozen sections and H/E stains. Sensibility, success index, negative predictive value, positive predictive value and false negative rate were calculated. RESULTS: Sentinel node was identified in 57/65 patients (87%) in blue dye group and in all the patients in combined technique group. There were sentinel node metastasis in 19/79 (24%), 4/19 patients had metastatic non-sentinel nodes without sentinel node metastasis (false negative rate = 17%), all the false negative sentinel nodes in the blue dye only group. CONCLUSIONS: Lymphatic mapping and sentinel node biopsy could avoid unnecessary axillary dissections in early stages of breast cancer; the combined technique reduces the false negative rate and increases the index of success. PMID- 15550223 TI - [Analgesic alternatives for the control of postoperatory pain in radical mastectomy]. AB - INTRODUCTION: The purpose of preventive analgesia based on regional blocks with local anesthetics is to avoid sharp and hard-to-control pain in the anterior side of thorax and axilla secondary to radical mastectomy. OBJECTIVE: Our aim was to compare the analgesic effectiveness of direct trans-operational block applied to nerves vs conventional analgesia using analgesic-antipiretic and antiinflammatory (AINES) drugs. MATERIAL AND METHODS: We designed a 1-year case-control study at the Hospital de Oncologia of the Instituto Mexicano del Seguro Social's (IMSS's) Centro Medico Nacional Siglo XXI (CMN-SXXI) in Mexico City. A total of 112 patients were mastectomized under general anesthesia: Thirty one patients received preventive analgesia (cases) by direct transoperational infiltration of brachial and intercostal nerves III-VI; bupivacaine 1 mg/kg and buprenorphine 0.005 mg/kg in 15 ml were distributed among appropriate nerves accompanied by a single dose of AINES in emersion stage; during anesthetic emersion, 81 patients received conventional analgesia (control), including endovenous AINES alone or combined, and buprenorphine 0.3 mg or nalbuphine 10 mg. Three evaluations employing analog visual scale were conducted: recovery (immediate post surgery); floor (mediate), and following day. RESULTS: We studied 110 women and two men. Demographic, laboratory, anesthetic, and surgical variables were similar in both groups (p = not significant [ns]). In no case was post-surgical pain reported. Among controls, slight pain was reported by 13 patients, moderate pain by 66, and severe pain was reported by two patients, whereas in patients returned to their rooms on the floor, slight pain was reported by 38, moderate by 42, and severe by one. Finally, on the following day all patients reported slight pain. Types of pain included 15 patients with somatic pain, 10 with neuropathic, and 56 patients with mixed pain (p <0.0001). CONCLUSIONS: Preventive analgesia by direct transoperational block on nerves turned out to be an effective alternative with low level of systemic morbidity, which must be considered to prevent moderate and severe pain syndromes that are difficult to control in thoracic oncological surgery. PMID- 15550224 TI - [Considerations on employment of the laser Nd:YAG in pneumology]. AB - Employment of the laser in medicine has demonstrated its effectiveness. Laser Nd:YAG renders good results for treatment of obstructive malignant and benign airway lesions. There are no statistical differences in frequency of these two kind of lesions and immediate results of laser therapy. Proper case selection and knowledge of their limitations and their advantages are important. Results of a series of 112 patients are presented here. PMID- 15550226 TI - [Appendiceal tumors. Clinicopathologic review of 5,307 appendectomies]. AB - INTRODUCTION: Appendix tumors are unusual, accounting for 0.4% of all gastrointestinal- tract malignancies. Although rare, the spectrum of malignant disease is complex and has led to confusion with regard to accurate description of the natural history of these tumors. Consequently, many errors in diagnosis and treatment have occurred. OBJECTIVE: Our aim was to analyze incidence and clinic-pathologic behavior of appendiceal tumors. METHODS: Our study type was retrospective, observational, longitudinal, and descriptive, and was conducted at a secondary-care-level hospital. From a single care center, a histopathologic database of 5,307 appendectomies, all appendiceal tumors, was identified and case notes were reviewed. Analysis of clinical presentation, histopathology, operation, and outcome is presented. RESULTS: During a 10-year period (5,307 appendectomies), 31 patients (0.58%) with appendiceal tumors were identified: 11 had carcinoid tumors, 18 benign, and two patients had malignant tumors. Acute appendicitis was the most common presentation (79.1%), and 20.3% were normal appendices. Patients with appendiceal tumor included 23 females and eight males with mean age of 48 years. Ninety seven percent of patients had preoperative diagnosis of acute appendicitis. CONCLUSIONS: Appendiceal tumors are uncommon and most commonly present as acute appendicitis. Macroscopic suspicion during surgery is the most important point to make the best decision. PMID- 15550228 TI - [Peripheral hereditary neuropathies: Charcot-Marie-Tooth types 1 and 2]. AB - OBJECTIVE: We report 14 cases of patients with Charcot-Marie-Tooth types 1 (CMT) and type 2 (CMT2). The objective of this study was to determine the diagnostic value of structural studies of peripheral nerve and striated muscle biopsies and to discuss correlations with clinical symptoms and electrophysiologic findings. MATERIAL AND METHODS: Nerve and muscle biopsies were obtained and processed according to standards methods for transmission electron microscopy. Diagnosis of CMT was based on clinical electrophysiologic findings. RESULTS: Our results demonstrated that myelinated fibers showed moderate-to- severe demyelinization and remyelinization, folding complex, tomacula formations, and presence of moderate vacuoles into myelin. The axonal cytoskeleton shows an important decrease of neurofilaments and microtubules in myelinated and unmyelinated nerves. Striated muscle showed different degenerative changes in Z disc. One important finding was presence of crystals inclusions in mitochondrion. In 11 cases, it was possible to find relevant changes in mitochondria-like hyperplasia. In this study, we had four cases with > 20 years of evolution; these cases showed relevant degenerative changes in nerve and muscle. Nerves presented great demyelination with damage in cytoskeleton of the axon, while fibers of muscle had necrosis and hyalinization. CONCLUSIONS: According to our results, it is possible to distinguish morphologically CMT subtypes, due to the fact that in CMT2 we found a 50% decrease of filaments and microtubules that were reduced in myelinic axons; this finding does not occur in CMT1. PMID- 15550227 TI - [The role of long-acting release (LAR) depot octreotide as adjuvant management of short bowel disease]. AB - INTRODUCTION: Short bowel syndrome (SBS) comprises sequelaes of nutrient, fluid, and weight loss that occur subsequent to greatly reduced functional surface area of small intestine. Maintenance of fluid and electrolyte homeostasis in patients with SBS is often difficult; additional intravenous (i.v.) fluid and electrolytes are often required to cover unabsorbed secretory losses. The recent development of the synthetic long-acting release (LAR) depot octapeptide analog, octreotide, has rendered somatostatin therapy a practical alternative in patients with SBS who have become entrapped in a vicious cycle with chronic dehydration, thirst, and excessive fluid or food consumption. METHODS: During an 8-month period, adult patients of either sex with SBS and total parental nutrition (TPN) dependent on III adaptation intestinal phase were included in a prospective, open-label study. We used single-subject research. Patients were divided into two groups: intestinal reinstated patients, group A (n = 6), and patients with high stomal output, group B (n = 4). Octreotide depot (20 mg intramuscularly [i.m.] at 0-8 months) was administered. Baseline and post-treatment measurements of nutritional status and stool losses were analyzed. RESULTS: Treatment with octreotide depot significantly reduced intestinal output (group A, p = 0.002, group B, p = 0.823), hospital stay, i.v. fluid and electrolyte requirements, in 80% TPN was not required. CONCLUSIONS: Octreotide depot is effective in diminishing i.v. fluid and electrolyte requirements by reducing stool losses in SBS patients. Multicenter studies with control group and larger sample size are required. PMID- 15550230 TI - [Sutureless thyroidectomy with local anesthesia plus conscious sedation. Two clinical cases]. AB - OBJECTIVE: We proposed a modification of thyroid surgery and have reported two operations carried out with sutureless surgery and local anesthesia. Our study setting was Unit 305 of the General Hospital of Mexico in Mexico City. With regard to study design, two patients are presented with sutureless thyroid surgery in what we believe is the first publication of its kind in Mexico and probably the first publication in the world that reports the combination of both sutureless technique and local anesthesia. MATERIAL AND METHODS: Patient number 1 is 41 years of age with a 10-year history. Fine-needle biopsy denoted nodular goiter but definitive diagnosis was Hashimoto thyroiditis, for which we performed subtotal resection. Patient number 2 is 46 years of age, and has a history of 1 year of thyroid mass; final diagnosis was colloid goiter. Lobectomy and isthmus resection was done. We proceeded under local anesthesia plus sedation with the patient conscious, in combination with no stitches for hemostasia using a tissue sealing system ligasure. RESULTS: Both patients had good results without postoperative complications. Surgical time was 120 min in first case and 71 min for the second. Patient 1 had transient hoarseness for 3 weeks, and the healing was spontaneous. CONCLUSION: Outcome was adequate in these two cases with local anesthesia and no stitches. PMID- 15550231 TI - [Abdominal pseudocyst in a patient with ventriculoperitoneal shunt. A case report]. AB - Since 1905, the abdominal cavity has been used for absorption of cerebrospinal fluid in patients with hydrocephalus. Among complications in its use is formation of abdominal pseudocysts. We describe the case of a patient with hydrocephalus who developed an abdominal pseudocyst. The main complaint of the patient was abdominal pain and fever. During physical examination, a 15-cm abdominal tumor was detected. The patient underwent abdominal surgery for excision of the pseudocyst and peritoneal shunt was relocated to right atrium. Recovery was uneventful. Incidence of abdominal pseudocyst ranges from 1-4.5%. Principal symptoms are related with intracranial hypertension. The patient usually requires surgical exploration to resolve the illness, and in presence of infection the shunt should be changed. PMID- 15550232 TI - [Tuberculous psoas abscess in a patient with HIV]. AB - Psoas abscess is an uncommon disease with varied etiology. The most common pathogen in primary abscess is Staphylococcus aureus. Crohn's disease is the most common cause of a secondary abscess. Recently, drug abuse and HIV infection have become important risk factors in identification of a patient with psoas abscess. Mycobacterium tuberculosis is considered an extremely rare cause of psoas abscess, but it is reported in HIV patients. Physical presentation is insidious; classical symptoms are pain and fever. Computed tomography (CT) is the most important tool for diagnosis. Preferred treatment is percutaneous drainage guided by CT. Open surgical drainage should be reserved if percutaneous drainage fails, if possible by extraperitoneal approach; in secondary abscess, the transperitoneal approach is preferred to correct the possible disease. PMID- 15550233 TI - [Persistent hyperinsulinemic hypoglycemia. Two case reports]. AB - We report two cases of infants who presented seizures and persistent hypoglycemia. In both infants, we established diagnosis of persistent hypoglycemia by hyperinsulinism (PHH); glycemic/insulinic ratio was > 0.3, and we found no pancreatic changes by means of other diagnostic procedures. The medical treatment consisted of hydrocortisone, diazoxide, and octreotide without response. Definitive treatment made was sub-total pancreatectomy; postoperatory evolution was satisfactory, and the histologic report showed changes in Langerhans islet of nesidioblastosis. We conclude that PHH and glucose/insulin ratio > 0.3 indicates that surgical treatment with pancreatectomy should not be delayed. PMID- 15550234 TI - [Mitral valve prolapse. A review]. AB - Primary mitral valve prolapse (MVP) or the disease known as MVP, consists of mixomatous degeneration of the mitral valve with systolic displacement of a portion or all of one or both mitral leaflets beyond mitral annulus into left atrium during systole, associated or not with mitral insufficiency. Prevalence in Mexican population is <2%. MVP behavior is benign unless associated with mitral insufficiency (MI, moderate-to-severe) or complications, or is associated with other syndromes. The major clinical feature of mitral valve prolapse syndrome is mid-to-late systolic clicks identified with auscultation of mitral valve. Echocardiography is usually employed for diagnosis and management. Two dimensional echocardiography displays one or both leaflets prolapsing behind mitral annulus and into left atrium in systole. In the majority of cases, MVP is harmless and does not cause symptoms nor does it need to be treated. In a small number of cases, it can cause severe mitral regurgitation and needs surgical treatment. Complications derived from MVP must be treated independently. Secondary MVP also consist of displacement of one or both valves toward atrium, but this is due to pathologies such as rheumatic heart disease, ischemic heart disease, or others. In these situations, treatment will be that of the underlying diseases. PMID- 15550235 TI - [Left atrial dissection. Review]. AB - The left atrial dissection is a rare complication occurring mainly after mitral surgery, thoracic trauma, myocardial infarction and infectious endocarditis. The clinical diagnosis can be confused with isolated prosthetic dysfunction, myocardial infarction. Its onset and, its form of presentation vary widely, concerning to the intensity and to the moment in which the symptoms appear. The most important data for its diagnostic are the antecedents and a new systolic murmur, although this last one can be absent. The transesophageal echocardiogram is the first choice for diagnosis since, the dissection can be missed by transthoracic echocardiogram. The treatment is surgical, two different types of surgical approaches have been reported. The prognosis depends on the extension of the dissection, tissue quality and associated complications. PMID- 15550236 TI - [Use and production of biomedical models in the field of audiology]. AB - Presently paper mentions the importance that has the use and the production of animal models in scientific research, the relevance that has its use to be able to make a good pursuit in the research, highlighting as well as to facilitate the reproductive of the study in question. Lastly it is mentioned the models more used in the field of the audiology. PMID- 15550237 TI - Chaperoned protein disaggregation--the ClpB ring uses its central channel. AB - In this issue of Cell, exploit a clever manipulation of the Hsp100 ring chaperone, ClpB, to gain some mechanistic and physiologic understanding of the action of this chaperone in mediating ATP-dependent disaggregation of protein aggregates that accumulate in the bacterial cytoplasm under severe heat shock conditions. PMID- 15550238 TI - SNARE filtering by dynamin. AB - Fission and fusion are the two elementary steps of membrane traffic. The mechanoenzyme dynamin acts at the fission step, but, in this issue of Cell, suggest an additional role of dynamin in preparing membranes for fusion. PMID- 15550239 TI - Making the right choice--long-range chromosomal interactions in development. AB - Schizosaccharomyces pombe has the remarkable potential to switch mating type as often as every generation, through selective interaction of an expressor locus with either of two transcriptionally silent donor loci. Recent results demonstrate that selection of the appropriate donor locus likely occurs through mating-type and heterochromatin-dependent spreading of a protein complex that marks the correct donor locus. PMID- 15550240 TI - Arginine methylation of STAT1: a reassessment. PMID- 15550242 TI - A single nucleotide polymorphism in the MDM2 promoter attenuates the p53 tumor suppressor pathway and accelerates tumor formation in humans. AB - The tumor suppressor p53 gene is mutated in minimally half of all cancers. It is therefore reasonable to assume that naturally occurring polymorphic genetic variants in the p53 stress response pathway might determine an individual's susceptibility to cancer. A central node in the p53 pathway is the MDM2 protein, a direct negative regulator of p53. In this report, a single nucleotide polymorphism (SNP309) is found in the MDM2 promoter and is shown to increase the affinity of the transcriptional activator Sp1, resulting in higher levels of MDM2 RNA and protein and the subsequent attenuation of the p53 pathway. In humans, SNP309 is shown to associate with accelerated tumor formation in both hereditary and sporadic cancers. A model is proposed whereby SNP309 serves as a rate limiting event in carcinogenesis. PMID- 15550243 TI - Methylation of histone H4 lysine 20 controls recruitment of Crb2 to sites of DNA damage. AB - Histone lysine methylation is a key regulator of gene expression and heterochromatin function, but little is known as to how this modification impinges on other chromatin activities. Here we demonstrate that a previously uncharacterized SET domain protein, Set9, is responsible for H4-K20 methylation in the fission yeast Schizosaccharomyces pombe. Surprisingly, H4-K20 methylation does not have any apparent role in the regulation of gene expression or heterochromatin function. Rather, we find the modification has a role in DNA damage response. Loss of Set9 activity or mutation of H4-K20 markedly impairs cell survival after genotoxic challenge and compromises the ability of cells to maintain checkpoint mediated cell cycle arrest. Genetic experiments link Set9 to Crb2, a homolog of the mammalian checkpoint protein 53BP1, and the enzyme is required for Crb2 localization to sites of DNA damage. These results argue that H4-K20 methylation functions as a "histone mark" required for the recruitment of the checkpoint protein Crb2. PMID- 15550244 TI - Structural basis of ligand activation in a cyclic nucleotide regulated potassium channel. AB - Here we describe the initial functional characterization of a cyclic nucleotide regulated ion channel from the bacterium Mesorhizobium loti and present two structures of its cyclic nucleotide binding domain, with and without cAMP. The domains are organized as dimers with the interface formed by the linker regions that connect the nucleotide binding pocket to the pore domain. Together, structural and functional data suggest the domains form two dimers on the cytoplasmic face of the channel. We propose a model for gating in which ligand binding alters the structural relationship within a dimer, directly affecting the position of the adjacent transmembrane helices. PMID- 15550245 TI - Molecular mechanism of AHSP-mediated stabilization of alpha-hemoglobin. AB - Hemoglobin A (HbA), the oxygen delivery system in humans, comprises two alpha and two beta subunits. Free alpha-hemoglobin (alphaHb) is unstable, and its precipitation contributes to the pathophysiology of beta thalassemia. In erythrocytes, the alpha-hemoglobin stabilizing protein (AHSP) binds alphaHb and inhibits its precipitation. The crystal structure of AHSP bound to Fe(II)-alphaHb reveals that AHSP specifically recognizes the G and H helices of alphaHb through a hydrophobic interface that largely recapitulates the alpha1-beta1 interface of hemoglobin. The AHSP-alphaHb interactions are extensive but suboptimal, explaining why beta-hemoglobin can competitively displace AHSP to form HbA. Remarkably, the Fe(II)-heme group in AHSP bound alphaHb is coordinated by the distal but not the proximal histidine. Importantly, binding to AHSP facilitates the conversion of oxy-alphaHb to a deoxygenated, oxidized [Fe(III)], nonreactive form in which all six coordinate positions are occupied. These observations reveal the molecular mechanisms by which AHSP stabilizes free alphaHb. PMID- 15550246 TI - Symplekin and xGLD-2 are required for CPEB-mediated cytoplasmic polyadenylation. AB - Cytoplasmic polyadenylation-induced mRNA translation is a hallmark of early animal development. In Xenopus oocytes, where the molecular mechanism has been defined, the core factors that control this process include CPEB, an RNA binding protein whose association with the CPE specifies which mRNAs undergo polyadenylation; CPSF, a multifactor complex that interacts with the near ubiquitous polyadenylation hexanucleotide AAUAAA; and maskin, a CPEB and eIF4E binding protein whose regulation of initiation is governed by poly(A) tail length. Here, we define two new factors that are essential for polyadenylation. The first is symplekin, a CPEB and CPSF binding protein that serves as a scaffold upon which regulatory factors are assembled. The second is xGLD-2, an unusual poly(A) polymerase that is anchored to CPEB and CPSF even before polyadenylation begins. The identification of these factors has broad implications for biological process that employ polyadenylation-regulated translation, such as gametogenesis, cell cycle progression, and synaptic plasticity. PMID- 15550247 TI - Thermotolerance requires refolding of aggregated proteins by substrate translocation through the central pore of ClpB. AB - Cell survival under severe thermal stress requires the activity of the ClpB (Hsp104) AAA+ chaperone that solubilizes and reactivates aggregated proteins in concert with the DnaK (Hsp70) chaperone system. How protein disaggregation is achieved and whether survival is solely dependent on ClpB-mediated elimination of aggregates or also on reactivation of aggregated proteins has been unclear. We engineered a ClpB variant, BAP, which associates with the ClpP peptidase and thereby is converted into a degrading disaggregase. BAP translocates substrates through its central pore directly into ClpP for degradation. ClpB-dependent translocation is demonstrated to be an integral part of the disaggregation mechanism. Protein disaggregation by the BAP/ClpP complex remains dependent on DnaK, defining a role for DnaK at early stages of the disaggregation reaction. The activity switch of BAP to a degrading disaggregase does not support thermotolerance development, demonstrating that cell survival during severe thermal stress requires reactivation of aggregated proteins. PMID- 15550248 TI - Mutual control of membrane fission and fusion proteins. AB - Membrane fusion and fission are antagonistic reactions controlled by different proteins. Dynamins promote membrane fission by GTP-driven changes of conformation and polymerization state, while SNAREs fuse membranes by forming complexes between t- and v-SNAREs from apposed vesicles. Here, we describe a role of the dynamin-like GTPase Vps1p in fusion of yeast vacuoles. Vps1p forms polymers that couple several t-SNAREs together. At the onset of fusion, the SNARE-activating ATPase Sec18p/NSF and the t-SNARE depolymerize Vps1p and release it from the membrane. This activity is independent of the SNARE coactivator Sec17p/alpha-SNAP and of the v-SNARE. Vps1p release liberates the t-SNAREs for initiating fusion and at the same time disrupts fission activity. We propose that reciprocal control between fusion and fission components exists, which may prevent futile cycles of fission and fusion. PMID- 15550249 TI - RTP family members induce functional expression of mammalian odorant receptors. AB - Transport of G protein-coupled receptors (GPCRs) to the cell surface membrane is critical in order for the receptors to recognize their ligands. However, mammalian GPCR odorant receptors (ORs), when heterologously expressed in cells, are poorly expressed on the cell surface. Here we show that the transmembrane proteins RTP1 and RTP2 promote functional cell surface expression of ORs expressed in HEK293T cells. Genes encoding these proteins are expressed specifically in olfactory neurons. These proteins are associated with OR proteins and enhance the OR responses to odorants. Similar although weaker effects were seen with a third protein, REEP1. These findings suggest that RTP1 and RTP2 in particular play significant roles in the translocation of ORs to the plasma membrane as well as in the functioning of ORs. We have used this approach to identify active odorant ligands for ORs, providing a platform for screening the chemical selectivity of the large OR family. PMID- 15550250 TI - Circadian gene expression in individual fibroblasts: cell-autonomous and self sustained oscillators pass time to daughter cells. AB - The mammalian circadian timing system is composed of a central pacemaker in the suprachiasmatic nucleus (SCN) of the brain and subsidiary oscillators in most peripheral cell types. While oscillators in SCN neurons are known to function in a self-sustained fashion, peripheral oscillators have been thought to damp rapidly when disconnected from the control exerted by the SCN. Using two reporter systems, we monitored circadian gene expression in NIH3T3 mouse fibroblasts in real time and in individual cells. In conjunction with mathematical modeling and cell co-culture experiments, these data demonstrated that in vitro cultured fibroblasts harbor self-sustained and cell-autonomous circadian clocks similar to those operative in SCN neurons. Circadian gene expression in fibroblasts continues during cell division, and our experiments unveiled unexpected interactions between the circadian clock and the cell division clock. Specifically, the circadian oscillator gates cytokinesis to defined time windows, and mitosis elicits phase shifts in circadian cycles. PMID- 15550251 TI - Independent regulation of synaptic size and activity by the anaphase-promoting complex. AB - Neuronal plasticity relies on tightly regulated control of protein levels at synapses. One mechanism to control protein abundance is the ubiquitin-proteasome degradation system. Recent studies have implicated ubiquitin-mediated protein degradation in synaptic development, function, and plasticity, but little is known about the regulatory mechanisms controlling ubiquitylation in neurons. In contrast, ubiquitylation has long been studied as a central regulator of the eukaryotic cell cycle. A critical mediator of cell-cycle transitions, the anaphase-promoting complex/cyclosome (APC/C), is an E3 ubiquitin ligase. Although the APC/C has been detected in several differentiated cell types, a functional role for the complex in postmitotic cells has been elusive. We describe a novel postmitotic role for the APC/C at Drosophila neuromuscular synapses: independent regulation of synaptic growth and synaptic transmission. In neurons, the APC/C controls synaptic size via a downstream effector Liprin-alpha; in muscles, the APC/C regulates synaptic transmission, controlling the concentration of a postsynaptic glutamate receptor. PMID- 15550252 TI - A behavioral role for dendritic integration: HCN1 channels constrain spatial memory and plasticity at inputs to distal dendrites of CA1 pyramidal neurons. AB - The importance of long-term synaptic plasticity as a cellular substrate for learning and memory is well established. By contrast, little is known about how learning and memory are regulated by voltage-gated ion channels that integrate synaptic information. We investigated this question using mice with general or forebrain-restricted knockout of the HCN1 gene, which we find encodes a major component of the hyperpolarization-activated inward current (Ih) and is an important determinant of dendritic integration in hippocampal CA1 pyramidal cells. Deletion of HCN1 from forebrain neurons enhances hippocampal-dependent learning and memory, augments the power of theta oscillations, and enhances long term potentiation (LTP) at the direct perforant path input to the distal dendrites of CA1 pyramidal neurons, but has little effect on LTP at the more proximal Schaffer collateral inputs. We suggest that HCN1 channels constrain learning and memory by regulating dendritic integration of distal synaptic inputs to pyramidal cells. PMID- 15550254 TI - Treatment of acute vivax malaria with tafenoquine. AB - Tafenoquine is an 8-aminoquiniline related to primaquine with pre-clinical activity against a range of malaria species. We treated two acute cases of vivax malaria with tafenoquine (800 mg over three days) alone, instead of conventional chloroquine (1500 mg over three days) and primaquine (420 mg over 14 days). In addition to the convenience of this regimen, the rapid parasite clearances observed, coupled with a good clinical response and lack of recrudescence or relapse, indicate that further investigation of tafenoquine in the treatment of vivax malaria is warranted. PMID- 15550255 TI - Simplification and improvement of height-based azithromycin treatment for paediatric trachoma. AB - Recent data showing that azithromycin is safe at higher dosages than previously documented provide an opportunity to explore several important improvements in the efficiency and effectiveness of height-based treatment of paediatric trachoma. The purpose of this study is to examine the feasibility of a single standardised schedule for application in any trachoma-endemic region. Data for 60813 children from Asia, North and sub-Saharan Africa were analysed. A height schedule maximizing the number of children receiving treatment of 20-40 mg/kg, a conservative estimate of the safe and effective treatment range for paediatric trachoma, was developed. Using the standardised schedule, 97.7% of children aged 6 to 59 months receiving oral suspension and 96.7% of children aged 60 months to 15 years receiving tablets would have received treatment within a dosage range of 20-40 mg/kg. Less than 1% of all children would have received treatment less than 20 mg/kg. These findings suggest that the schedule presented in this paper is likely to yield safe and effective treatment for a broad range of populations vulnerable to trachoma while substantially improving the efficiency of height based treatment. PMID- 15550256 TI - Atypical cutaneous leishmaniasis in Central America: possible interaction between infectious and environmental elements. AB - Biopsies of 71 cases of atypical cutaneous leishmaniasis from Costa Rican patients were evaluated by histopathological procedures and attempts were made to culture Leishmania from nine biopsies. Leishmanin skin tests were carried out in 31 patients and 112 healthy individuals. Additional biopsies from 19 patients from Nicaragua were evaluated by routine histopathology. Ten biopsies were studied by confocal and nine by scanning electron microscopy. Inorganic material was analysed using an electron probe for microanalysis. Leishmania parasites were isolated from only two biopsies, but 90.3% of the patients from Costa Rica were leishmanin-positive, as were 27.7% of healthy individuals. Routine histopathological studies revealed naked granulomas formed by differentiated macrophages. Abundant inorganic material was observed in sections examined by confocal microscopy. Electron probe analysis revealed that silica and aluminium were the predominant elements in large particles. We postulate that the presence of this inorganic material, possibly of volcanic origin, in the skin may modulate the immunological response to Leishmania and may inhibit visceralization in the cases caused by Leishmania chagasi. PMID- 15550257 TI - Epidemiology of helminth infections and their relationship to clinical malaria in southwest Uganda. AB - It has recently been suggested that helminth infections may adversely influence susceptibility to other infections, including malaria. To investigate this hypothesis in a sub-Saharan African setting, surveys of helminth infections were conducted in 2003 among individuals who had been under weekly active case detection for clinical malaria during the preceding 18 months in four villages in Kabale District, southwest Uganda. Overall, 47.3% of individuals had at least one intestinal nematode species infection: hookworm, Ascaris lumbricoides and Trichuris trichiura were detected in 32.1, 17.4 and 8.1% of individuals, respectively. We found evidence of significant household clustering of A. lumbricoides, T. trichiura and hookworm, and clustering of heavy infection of each species. The association between helminth infection and clinical malaria was investigated in two villages and no evidence for an association was observed between the presence of infection or heavy infection and risk of malaria. PMID- 15550258 TI - A new PCR-ELISA for diagnosis of visceral leishmaniasis in blood of HIV-negative subjects. AB - The PCR-ELISA represents a promising advance for diagnosis of visceral leishmaniasis (VL) in blood samples. However, the method has been validated mostly with HIV-positive patients who are known to have high levels of parasitaemia. We developed a new PCR-ELISA assay for specific detection of Leishmania in patients' blood and validated it in Nepalese subjects with clinically suspected VL, almost all of whom were HIV-negative. For blood samples, PCR-ELISA was more sensitive (83.9%) than conventional PCR (73.2%), and demonstrated 100% and 87.2% specificity when using healthy controls who had never travelled to a VL-endemic area and controls from a VL-endemic area as references, respectively. We have demonstrated the ability of PCR-ELISA to detect parasites in blood of HIV-negative patients. The method could be used for epidemiological as well as clinical purposes, as it reduces the need for traumatic bone marrow sampling and risky spleen aspiration. PMID- 15550259 TI - Distribution of human Oesophagostomum bifurcum, hookworm and Strongyloides stercoralis infections in northern Ghana. AB - A cross-sectional study was carried out in 216 randomly selected, representative rural villages in the northeastern part of Ghana from March 1995 to May 1998. Inhabitants of randomly selected households, stratified by age and gender, were included. The geographical position of villages was recorded with a global positioning system (GPS). The prevalence of Oesophagostomum, hookworm and Strongyloides stercoralis infections in a study population of 20250 people was determined by microscopic examination of larvae in stool cultures. The overall prevalence was 10.2, 50.6 and 11.6% for the three nematodes, respectively. Hookworm infections were seen in all but one (99.5%) and S. stercoralis in 88.4% of the 216 villages, while Oesophagostomum infections were found to be common in a limited area with prevalences varying from 0 to 75%. An association was found between Oesophagostomum and hookworm infection, both at the individual and at the village level. Spatial analysis of the prevalence data indicated that the endemic area is relatively clearly demarcated to the south of the study area. PMID- 15550260 TI - Definition of an algorithm for the management of common skin diseases at primary health care level in sub-Saharan Africa. AB - In order to help primary health care (PHC) workers in developing countries in the care of common skin diseases, an algorithm for the management of pyoderma, scabies, superficial mycoses, contact dermatitis and referral of early leprosy cases (based on the identification of diseases through the presence of objective key signs, and on treatments by generic drugs) was elaborated. One thousand patients were seen by trained dermatologists, who established diagnoses and treatments; in addition, there was systematic recording of each key sign, according to the successive algorithm steps. We compared the diagnostics and treatments obtained for several combinations of diagnostic signs, with those of the dermatologists. Sensitivity, specificity, positive predictive value and negative predictive value of defined combinations were high for pyoderma, scabies and superficial mycoses. Values were less exact for dermatitis and leprosy, but were considered sufficient for the level of health care targeted. The apportionment of treatments between the algorithm and the dermatological approaches was considered appropriate in more than 80% of cases; mismanagement was possible in 7% of cases, with few predictable harmful consequences. The algorithm was found satisfactory for the management of the dermatological priorities according to the standards required at the PHC level. PMID- 15550261 TI - Diarrhoea and garbage disposal in Salvador, Brazil. AB - The association of infantile diarrhoea with the presence of garbage in the environment was investigated in Canabrava, a peripheral neighbourhood of Salvador, northeast of Brazil. A cross-sectional study was conducted with all the 184 children aged less than two years residing in Canabrava, which is located close to the city garbage dump. Variables selected for study included the method used for the disposal of excrement, type of floor, mother's education, unemployment of the head of the family, regularity of the water supply, presence of toilet, storage of garbage inside the house, age, gender, duration of breastfeeding, and the number of people per room. The estimated prevalence of diarrhoea was 21.2%. Exposure to garbage in the environment was found to be the most important factor associated with diarrhoea (adjusted odds ratio [AOR] = 3.98, 95% CI 1.56-10.13). Other important variables were the mother's education (AOR = 2.79, 95% CI 1.09-7.13), maternal breastfeeding (AOR = 2.30, 95% CI 1.05 5.04), and unemployment of the head of the family (AOR = 2.09, 95% CI 0.93-4.69). These findings indicate the necessity of adopting solutions in the public domain and of intersectorial policies for the reduction of diarrhoea. PMID- 15550262 TI - Risk factors for mucosal manifestation of American cutaneous leishmaniasis. AB - A case-comparison study was carried out to identify risk factors for mucosal manifestations of American cutaneous leishmaniasis (ACL) in southeast Brazil, using a series of 2820 patients, diagnosed with ACL between 1966 and 1999. The significant factors independently associated with mucosal leishmaniasis were: gender, age, nutritional status and length of disease. Mucosal leishmaniasis occurred 1.7 times more frequently among males than females; twice as often in individuals older than 22 years compared with the younger group; almost four times as often in individuals with severe malnutrition compared with those who were well nourished; and almost four times more frequently in individuals reporting the disease for more than 4 months compared with those reporting a shorter duration of the disease. Among individuals older than 22 years the risk of mucosal leishmaniasis increased significantly (from 1.9 to 9.6) as the nutritional status decreased, when compared with younger and well-nourished patients. The characteristics herein described and correlated with severe forms could be used as diagnostic markers as part of clinical screening in areas endemic for ACL. PMID- 15550263 TI - In vitro recrudescence of Plasmodium falciparum parasites suppressed to dormant state by atovaquone alone and in combination with proguanil. AB - We studied the viability of Plasmodium falciparum parasites reappearing in long term cultures after repetitive exposure to atovaquone and proguanil. Parasites (F32 and FCR3) exposed to 100-5000 nM atovaquone for 96 hours were reduced to <5% of initial parasitaemia but recrudesced after 9-15 days. Also, parasites exposed to 1000 nM atovaquone for 48, 72, 96 and 144 hours recrudesced after 9, 14, 21 and 23 days respectively. Immediately after removal of the drug, only 1-3 schizonts per 10000 red blood cells were found consistently, apparently unable to produce trophozoites and thus, possibly, adopting a "dormant state". Parasites (F32 and FCR3) exposed to 500 nM atovaquone for 72 hours reappeared after 14 days. These recrudescing parasites were then re-exposed and suppressed by atovaquone in three consecutive follow-up experiments. They reappeared after 12, 11 and 9 days respectively. No known point mutations in cytochrome b gene (cytb), associated with atovaquone resistance, were detected in any recrudescing parasites. Finally, parasites (F32) exposed to various concentrations of atovaquone and proguanil in combination for 72 hours reappeared after 9-17 days. The baseline susceptibilities of the parasites to individual drugs were similar before and after recrudescence in all experiments. PMID- 15550265 TI - Physicians' management of filarial lymphoedema and hydrocele in Pondicherry, India. AB - Many countries have implemented mass drug administration programmes to eliminate lymphatic filariasis, but few have also implemented morbidity management programmes to help patients with chronic lymphoedema or hydrocele due to the infection. A study was carried out to assess current morbidity management by physicians in Pondicherry, India. Fifty-two physicians were interviewed, using a semi-structured questionnaire. For the management of hydrocele, all physicians referred hydrocele patients for surgery and 83% prescribed diethylcarbamazine (DEC). For the management of chronic lymphoedema patients, most doctors (75%) prescribed DEC and 56% mentioned the possibility of surgery. Only 10% of the physicians gave advice about limb hygiene, although recent research has shown the importance of hygiene measures to prevent further progression of lymphoedema. For the management of lymphoedema patients presenting with acute attacks, all physicians prescribed DEC and antibiotics and only 15% gave advice about limb hygiene. We conclude that limb hygiene instruction for home care should be more strongly emphasised to optimize management of lymphoedema patients in Pondicherry. PMID- 15550264 TI - Travel as a risk factor for uncomplicated Plasmodium falciparum malaria in the highlands of western Kenya. AB - In the 1980s, highland malaria returned to the tea estates of western Kenya after an absence of nearly a generation. In order to determine the importance of travel for the spread of malaria in this region, we prospectively collected blood films and travel, demographic and geographic information on well persons and outpatients on tea estates near the western rim of the Rift Valley. Risk factors for malaria asexual parasitaemia included: tribal/ethnic group, home province and home district malaria endemicity. Travel away from the Kericho tea estates within the previous two months showed an odds ratio (OR) for parasitaemia of 1.59 for well persons and 2.38 for outpatients. Sexual stages of malaria parasites (gametocytes) had an OR of 3.14 (well persons) and 2.22 (outpatients) for those who had travelled. Increased risk of malaria parasitaemia with travel was concentrated in children aged <5 years. An increase in population gametocytaemia is possibly due to increased chloroquine resistance and suppressed infections contracted outside of the tea estates. PMID- 15550266 TI - The WHO dose pole for the administration of praziquantel is also accurate in non African populations. AB - In 2001, WHO developed a pole for the administration of praziquantel without the use of weighing scales, with encouraging results in African populations. In the present study, the pole was tested on height/weight data from 9354 individuals from 11 non-African countries. In more than 98% of the individuals (95% CI 97.8 98.4) the pole estimated an acceptable dosage (30-60 mg/kg), a performance statistically similar to that observed in African populations. Reproducing the present pole in the form of a strip of paper and including it in each container of praziquantel would greatly facilitate the administration of the drug in large scale interventions. PMID- 15550267 TI - Splenic infarction during acute malaria. AB - Malaria is the most frequent cause of fever among travellers returning from tropical countries. Each year about 7000 cases are notified in France, of which 90% are due to Plasmodium falciparum. We describe the case of a Caucasian female patient with no previous exposure to malaria in whom splenic infarction occurred during effective antimalarial treatment for initially uncomplicated acute malaria. Management was restricted to close clinical monitoring and analgesia (subcutaneous morphine). Imaging abnormalities resolved within a few months. We found seven other such cases in the literature. All seven patients were younger and splenic infarction occurred later than in the case we describe. Clinical outcome was favourable in all the cases. It is noteworthy that this rare complication can occur despite appropriate antimalarial prophylaxis and treatment. There are no known predictive signs. Clinicians must be aware that left hypochondrial pain occurring during treatment for acute malaria may be due to splenic infarction. PMID- 15550268 TI - Crimean-Congo hemorrhagic fever. AB - Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne disease caused by the arbovirus Crimean-Congo hemorrhagic fever virus (CCHFV), which is a member of the Nairovirus genus (family Bunyaviridae). CCHF was first recognized during a large outbreak among agricultural workers in the mid-1940s in the Crimean peninsula. The disease now occurs sporadically throughout much of Africa, Asia, and Europe and results in an approximately 30% fatality rate. After a short incubation period, CCHF is characterized by a sudden onset of high fever, chills, severe headache, dizziness, back, and abdominal pains. Additional symptoms can include nausea, vomiting, diarrhea, neuropsychiatric, and cardiovascular changes. In severe cases, hemorrhagic manifestations, ranging from petechiae to large areas of ecchymosis, develop. Numerous genera of ixodid ticks serve both as vector and reservoir for CCHFV; however, ticks in the genus Hyalomma are particularly important to the ecology of this virus. In fact, occurrence of CCHF closely approximates the known world distribution of Hyalomma spp. ticks. Therefore, exposure to these ticks represents a major risk factor for contracting disease; however, other important risk factors are known and are discussed in this review. In recent years, major advances in the molecular detection of CCHFV, particularly the use of real-time reverse transcription-polymerase chain reaction (RT-PCR), in clinical and tick samples have allowed for both rapid diagnosis of disease and molecular epidemiology studies. Treatment options for CCHF are limited. Immunotherapy and ribavirin have been tried with varying degrees of success during sporadic outbreaks of disease, but no case-controlled trials have been conducted. Consequently, there is currently no antiviral treatment for CCHF approved by the U.S. Food and Drug Administration (FDA). However, renewed interested in CCHFV, as well as increased knowledge of its basic biology, may lead to improved therapies in the future. This article reviews the history, epidemiology, ecology, clinical features, pathogenesis, diagnosis, and treatment of CCHF. In addition, recent advances in the molecular biology of CCHFV are presented, and issues related to its possible use as a bioterrorism agent are discussed. PMID- 15550269 TI - Isolation and characterization of herpes simplex virus type 1 resistant to aminothiazolylphenyl-based inhibitors of the viral helicase-primase. AB - The aminothiazolylphenyl-containing compounds BILS 179 BS and BILS 45 BS are novel inhibitors of the herpes simplex virus helicase-primase with antiviral activity in vitro and in animal models of HSV disease. To verify the mechanism of antiviral action, resistant viruses were selected by serial passage or by single step plaque selection of HSV-1 KOS in the presence of inhibitors. Three resistant isolates K138r3, K22r5, and K22r1 were found to be 38-, 316-, and 2500-fold resistant to BILS 22 BS, a potent analog of BILS 45 BS. All three viruses had growth properties in vitro similar to wild-type HSV-1 KOS but they were sensitive to acyclovir. Cutaneous and intra-cerebral inoculation of mice with K22r1 or K22r5 resulted in pathogenicity equivalent to that of HSV-1 KOS. Both isolates were fully competent for reactivation from latency following corneal inoculation. Helicase-primase purified from cells infected with resistant viruses showed decreased inhibition in an in vitro DNA-dependent ATPase assay that correlated well with antiviral resistance. Marker transfer experiments and DNA sequence analysis identified single base pair mutations clustered in the N-terminus of the UL5 gene that resulted in single amino acid changes in the UL5 protein. Taken together, the results indicate that helicase-primase inhibitors prevent HSV growth by inhibiting HSV helicase-primase through specific interaction with the UL5 protein. PMID- 15550270 TI - Effect of increasing thiolation of the polycytidylic acid strand of poly I:poly C on the alpha, beta and gamma interferon-inducing properties, antiviral and antiproliferative activities. AB - Double-stranded RNAs induce interferons and cause the development of antiviral and antiproliferative activities. Antiviral activity is related to the production of interferons and other proteins that stimulate various immunologic activities, which appear to contribute to their overall antiproliferative activity. The most active double-stranded RNA, polyI:polyC, was shown to be too toxic for therapeutic use. We conducted selective thiolation of the polyC strand at the five position of the cytosine bases, generating a partially thiolated polyC (MPC) which after annealing with a complimentary unmodified polyI, gave the thiolated double-stranded RNA, pI:MPC. We have explored antiviral and antiproliferative activities at various levels of thiolation and found that optimal responses can be obtained at 7.4% level of thiolation. This compound deserves further study of antiviral and antiproliferative responses in vivo, and eventually clinical exploration. Earlier studies have shown that this and related compounds are active against HIV-1, in human cells, and against DNA polymerases of DNA and RNA tumor viruses. PMID- 15550271 TI - A novel model of HPV infection in meshed human foreskin grafts. AB - The present study describes a novel meshing procedure that provided successful low-risk papillomavirus propagation and reproducible wart induction in human foreskin xenografts. The initial HPV6 and/or 11 inocula were collected from clinically excised human wart tissues and confirmed to be free of HPV16, 18 and 31 by PCR analysis. Human foreskin grafts were collected from a circumcision clinic, and pre-inoculated with HPV virions by scarification. Meshing was carried out with a Zimmer Skin Grafter Mesher. Grafts were cut to appropriate size (1cm x 1cm or 5mm x 5mm) for cutaneous or subcutaneous grafting to NIH-nu-bg-xid mice under halothane anesthesia. Cutaneous xenografts were dressed with antibiotics and protective band-aids for 3 weeks. In the paralleled experiment using the same viral stock containing both HPV6 and 11, and matched grafts, no visible papillomas were observed in non-meshed cutaneous xenografts (n = 4 up to 6 months). In comparison, six of eight cutaneous xenografts treated with the meshing procedure formed visible papillomas within 4 months. This high frequency of distinct papilloma induction over the surface of meshed xenografts were reproduced in subsequent experiments with viral stocks containing both HPV11 and 6 (8 out of 10 grafts), or with a single-type HPV11 inoculum (80-100%). In contrast, an initial viral stock of single-type HPV6 provided lower frequency and more delayed papilloma induction. Serial passage of HPV6 in the meshed xenograft appeared to improve both the induction frequency and growth rate up to the 3rd generation. Histology, in situ hybridization, and immunohistochemical analysis revealed similarity of xenograft warts to those observed in the clinic. The highly reproducible papilloma induction rate and successful viral stock propagation associated with the meshing procedure provide a novel feature in the HPV xenograft model. PMID- 15550272 TI - Xanthohumol, a novel anti-HIV-1 agent purified from Hops Humulus lupulus. AB - Xanthohumol, prenylchacone flavonoid, is a natural product with multi biofunctions purified from Hops Humulus lupulus. Its anti-HIV-1 activity was tested in the present study. Results showed that xanthohumol inhibited HIV-1 induced cytopathic effects, the production of viral p24 antigen and reverse transcriptase in C8166 lymphocytes at non-cytotoxic concentration. The EC50 values were 0.82, 1.28 and 0.50 microg/ml, respectively. The therapeutic index (TI) was about 10.8. Xanthohumol also inhibited HIV-1 replication in PBMC with EC50 value of 20.74 microg/ml. The activity of recombinant HIV-1 reverse transcriptase and the HIV-1 entry were not inhibited by xanthohumol. The results from this study suggested that xanthohumol is effective against HIV-1 and might serve as an interesting lead compound. It may represent a novel chemotherapeutic agent for HIV-1 infection. However, the mechanism of its anti-HIV-1 effect needs to be further clarified. PMID- 15550273 TI - Inhibition of bovine viral diarrhea virus (BVDV) by mizoribine: synergistic effect of combination with interferon-alpha. AB - Bovine viral diarrhea virus (BVDV) is a well-characterized member of the Flaviviridae family. BVDV may be a surrogate model for human hepatitis C virus (HCV), since HCV does not replicate efficiently in cell cultures and animals. Mizoribine, a nucleoside analog clinically used as an immunosuppressant, was found to be active against the replication of BVDV in cell culture. We further investigated the combination of mizoribine and interferon (IFN)-alpha for antiviral activity and found that the combination synergistically inhibited BVDV replication in bovine kidney cells, as monitored by the inhibition of virus induced cytopathicity. The combination of ribavirin and IFN-alpha was also synergistic in inhibiting BVDV replication. Treatment of infected cells with a combination of mizoribine and IFN-alpha at the concentrations, at which the respective compounds proved to be inactive, markedly reduced viral infectivity in culture supernatants. These results indicate that mizoribine in combination with IFN-alpha may have potential for the treatment of HCV infection. PMID- 15550274 TI - Promotion of oxidative stress in the aquatic macrophyte Ceratophyllum demersum during biotransformation of the cyanobacterial toxin microcystin-LR. AB - Cyanobacterial toxins have been shown to have adverse effects on mammals, birds and fish and are therefore being increasingly recognised as a potent stress and health hazard factor in aquatic ecosystems. Microcystins, which are cyclic heptapeptides and a main group of the cyanotoxins, are mainly retained within the producer-cells during cyanobacterial bloom development. However, these toxins are released into the surrounding medium by senescence and lysis of the blooms. The released toxins could then come into contact with a wide range of aquatic organisms including invertebrates, fish and aquatic plants. In many organisms, biotransformation of the toxins will take place via several glutathione-related conjugate. During the biotransformation process in which the toxin and the toxin conjugate are broken down, the formation of reactive oxygen species might occur. These reactive oxygen species activate several antioxidant enzymes such as superoxide dismutase, catalase, ascorbate peroxidase and also influence the glutathione-ascorbate cycle. Aim of this study was to investigate formation of the glutathione-conjugate, activation of glutathione S-transferases and the elevation of several antioxidant enzymes giving evidence for the promotion of oxidative stress by microcystins. During exposure of Ceratophyllum demersum to the cyanobacterial toxin microcystin-LR in an concentration of 5.0 microg/L, an elevation of microsomal and cytosolic glutathione S-transferase was measured, showing the beginning formation of the glutathione-toxin conjugate. The superoxide dismutase as well as in parallel the hydrogen peroxide level increased giving evidence for oxidative stress in the aquatic plant. Other reactive oxygen detoxifiying enzymes were also elevated and the glutathione pool, expressed in reduced glutathione and glutathione disulfide concentration was changed accordingly. PMID- 15550275 TI - Tissue-specific Cu bioaccumulation patterns and differences in sensitivity to waterborne Cu in three freshwater fish: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio), and gibel carp (Carassius auratus gibelio). AB - Rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio) and gibel carp (Carassius auratus gibelio) were exposed to copper (1-20 microM) in softened Antwerp City tap water at pH 7.3 +/- 0.1 and with a water hardness of 292.4 +/- 8.1 mg/L CaCO(3) (Ca 100.8 +/- 3.0 mg/L; Mg 11.0 +/- 0.2 mg/L). LC50s (96 h) were determined and copper accumulation in gills, liver, and kidney assessed over a 10 day period. Rainbow trout (96 h LC50: 3.3 microM/210 microg/L) were three times more sensitive to Cu exposure than common carp (96 h LC50: 10.4 microM/661 microg/L) and almost seven times more sensitive than gibel carp (96 h LC50: 22.0 microM/1398 microg/L). After 96 h, the incipient lethal level (ILL) was reached for common carp, and by the end of the experiment (>120 h) also for rainbow trout. The ILL was never reached for gibel carp. Survival analysis confirmed the differences in sensitivity shown by the 96 h LC50 values. At 1 microM Cu, the relative risk to die was six to seven times greater for rainbow trout as for common or gibel carp, respectively, while it was 9000 and 19,000 times greater at 5 microM Cu. Only the environmental Cu concentrations contributed significantly (P < 0.001) to the Time-To-Death (TTD). Tissue Cu concentrations did not relate to TTD. Among species, a clear difference in metal handling was apparent, with high liver residues and liver accumulation rates for the most sensitive species, the rainbow trout, and lower liver but higher kidney residues and kidney accumulation rates for the most resistant species, the gibel carp. Gill concentrations and accumulation rates were lowest in the sensitive rainbow trout. PMID- 15550276 TI - Comparative teratogenicity of chlorpyrifos and malathion on Xenopus laevis development. AB - The embryotoxic potential of chlorpyrifos (CPF) and malathion (MTN), two organophosphorus insecticides (OPs), was evaluated by modified Frog Embryo Teratogenesis Assay-Xenopus (FETAX). CPF and MTN were not embryolethal even at the highest concentration tested (6000 microg/l), but both exhibited a powerful teratogenicity. The probit analysis of malformed larva percentages showed a TC(50) of 161.54mug/l for CPF, and a TC(50) of 2394.01 microg/l for MTN. Therefore, CPF teratogenicity was about 15 times higher than MTN. Larvae of both exposed groups were mainly affected by ventral and/or lateral tail flexure coupled with abnormal gut coiling. Histopathological diagnosis displayed abnormal myotomes and myocytes with marked hypertrophies localized at the cell extremity, probably due to a break away of myofibril extremities at the intersomitic junction level. We speculate that this muscular damage was related to inhibition of acetylcholinesterase that showed a clear concentration-response in CPF and MTN exposed larvae. The teratogenic effects of these anti-cholinesterase compounds on Xenopus laevis myogenesis suggest a possible role played by OPs on induction of congenital muscular dystrophy. PMID- 15550277 TI - Arsenic availability, toxicity and direct role of GSH and phytochelatins in As detoxification in the green alga Stichococcus bacillaris. AB - Accumulation and toxicity of inorganic arsenic forms As(III) and As(V) to the green microalga Stichococcus bacillaris depended on environmental variables. pH of exposure and to a lesser extent elevated concentrations of humic acid, chloride and orthophosphate ions affected arsenic accumulation and its toxicity. As(V) was more toxic than As(III), especially at the near neutral pH 6.8. Intracellular As(V) uptake by algal cells was greater at pH 6.8 than at 8.2. In response to As(III) and As(V) the alga produced phytochelatins (PC(2-3)), but at As(V) exposure, their levels in cells were higher than with As(III), suggesting higher As(V) than As(III) availability and uptake. Arsenic in algal cells occurred in various complexes with non-protein SH groups. Some of these complexes dissociated under acidic conditions, but others were able to dissociate only at an alkaline pH. The former consisted of SH groups of phytochelatins. Those dissociating at an alkaline pH involved SH groups from both glutathione (GSH) and phytochelatins (PC) or their derivatives. In the predominant acid-stable mixed As SH complex, the ratio of SH (PC(2)) to SH (GSH) was 2:1, which suggests that one molecule of PC(2) (containing two SH groups) together with one molecule of GSH were involved in intracellular complexation of each As atom. This is the first demonstration of GSH involvement in arsenic complexation, in vivo. The intracellular concentration of As was greater than that of non-protein SH groups which suggests that not all the arsenic in algal cells was complexed and detoxified by thiol groups. PMID- 15550278 TI - Altered social behavior and sexual characteristics in mosquitofish (Gambusia holbrooki) living downstream of a paper mill. AB - Several environmental contaminants have been found in paper mill effluent including substances with androgenic properties. The presence of androgenic substances in paper mill effluent was also indicated in the present study which demonstrated masculinized anal fins of female mosquitofish from the paper mill effluent contaminated Fenholloway River. In addition, when compared to the nearby Econfina River, which does not receive paper mill effluent, fewer females from the contaminated river were pregnant, they were smaller and their estradiol concentration exhibited greater variation. Males from the Fenholloway River and the Econfina River had similar sperm counts, but the testes were larger and greater variation in testosterone concentration was observed in male fish from the Fenholloway River. In males and females from the Fenholloway River, liver weights were increased and computer-aided behavior analysis demonstrated a reduction in their social behavior when compared to reference fish. We conclude that a number of sexual characteristics were affected in mosquitofish living in the paper mill contaminated Fenholloway River, with possible adverse effects on the reproduction of this population. PMID- 15550279 TI - Manganese induced immune suppression of the lobster, Nephrops norvegicus. AB - Manganese (Mn) is one of the most abundant elements on earth, particularly in the soft bottom sediments of the oceans. As a micronutrient Mn is essential in the metabolic processes of organisms. However, at high concentrations the metal becomes a neurotoxin with well-documented effects. As a consequence of euthrophication, manganese is released from bottom sediments of coastal areas and the Norway lobsters, Nephrops norvegicus, can experience high levels of bioavailable Mn(2+). Here, we present the first report showing that Mn also affects several fundamental processes in the mobilisation and activation of immunoactive haemocytes. When N. norvegicus was exposed to a realistic [Mn(2+)] of 20 mg l(-1) for 10 days 24.1 microg ml(-1) was recorded in the haemolymph. At this concentration the total haemocyte count was reduced by ca. 60%. By using BrdU as a tracer for cell division, it was shown that the proliferation rate in the haematopoietic tissue did not increase, despite the haemocytepenia. A gene coding for a Runt-domain protein, known to be involved in maturation of immune active haemocytes in a variety of organisms, was identified also in haemocytes of N. norvegicus. The expression of this gene was >40% lower in the Mn-exposed lobsters as judged by using a cDNA probe and the in situ hybridisation technique. In response to non-self molecules, like lipopolysaccharide (LPS), the granular haemocytes of arthropods are known to degranulate and thereby release and activate the prophenoloxidase system, necessary for their immune defence. A degranulation assay, tested on isolated granular haemocytes, showed about 75% lower activity in the Mn-exposed lobsters than that for the unexposed. Furthermore, using an enzymatic assay, the activation per se of prophenoloxidase by LPS was found blocked in the Mn-exposed lobsters. Taken together, these results show that Mn exposure suppressed fundamental immune mechanisms of Norway lobsters. This identifies a potential harm that also exists for other organisms and should be considered when increasing the distribution of bioavailable Mn, as has been done through recently introduced applications of the metal. PMID- 15550280 TI - Reproductive toxicity of dietary zinc to Daphnia magna. AB - Regulatory assessments of metals in freshwaters are mostly based on dissolved metal concentrations, assuming that toxicity is caused by waterborne metal only. Little attention has been directed to the toxicity of dietary metals to freshwater invertebrates. In this study the chronic toxicity of dietary zinc to Daphnia magna was investigated. The green alga Pseudokirchneriella subcapitata was exposed for 64 h to a control and three dissolved zinc concentrations, i.e. 23, 28 and 61 microg L(-1), resulting in internal zinc burdens in the algae of 130, 200, 320 and 490 microg g(-1) dry weight, respectively. These algae were used as a food source in chronic, 21-day bioassays with D. magna in a test medium to which no dissolved zinc was added. None of the treatments resulted in effects on feeding rates or somatic growth of D. magna. In contrast, a significant 40% decrease of total reproduction (number of juveniles per adult) was observed in the 28 and 61 microg L(-1) treatments. Time to first brood was not affected, whereas the mean brood size and the fraction of reproducing parent daphnids were reduced from the second brood onwards and the magnitude of these reductions increased with each subsequent brood. The reduced reproduction was accompanied with an elevated zinc accumulation in the 61 microg L(-1) treatment only, suggesting that total body burden is no good indicator of dietary zinc toxicity. Overall our data suggest that dietary zinc specifically targets reproduction in D. magna through accumulation in particular target sites, possibly cells or tissues where vitellogenin synthesis or processing occur. Further, our data illustrate that the potential importance of the dietary exposure route should be carefully considered and interpreted in regulatory assessments of zinc. PMID- 15550281 TI - Temperature influences on the accumulation and elimination of mercury in a freshwater cladoceran, Daphnia magna. AB - Temperature is an important environmental variable affecting the physiology and metabolism of aquatic invertebrates and can potentially affect the rate and pathway of the uptake and elimination of contaminants. In this study, we studied the effects of the ambient temperature on the uptake (from water and food) and elimination of inorganic mercury [Hg(II)] and methymercury (MeHg) in a freshwater cladoceran, Daphnia magna, in the laboratory. At different temperatures (i.e., 14, 19 and 24 degrees C), the assimilation efficiencies of both Hg species from ingested alga were not significantly different while the dissolved uptake rates were 32 and 73% lower at 14 degrees C than at 24 degrees C for Hg(II) and MeHg, respectively. The reduced uptake rates of Hg(II) at 14 degrees C could be adequately explained by the reduced filtration rate (40% reduction), but for MeHg, the reduced filtration rate could not completely explain reduction in MeHg uptake rate. Despite the elimination rates of both Hg compounds not being affected by the temperature, the relative importance of different routes of Hg loss (i.e., excretion, egestion, molting and reproduction) was significantly different at various temperatures. The relative importance of excretion to MeHg loss increased from 24 to 14 degrees C (i.e., 52 to 85% of Hg loss) while that of reproduction to MeHg loss decreased from 43 to 11% simultaneously. Using a kinetic model, we showed that the fraction of MeHg accumulated in the daphnids coming from the dietary phase was higher at lower temperature, while there was increased accumulation of both Hg(II) and MeHg in the daphnids at higher temperatures. Nevertheless, the trophic transfer potentials for both Hg species in this algae-daphnids food chain were not influenced by the temperature. PMID- 15550282 TI - Addictive disorders. AB - In conclusion, complex medical and psychiatric comorbidity is com-mon in individuals with substance use disorders. It is important to assess comorbidity because of the implications for prevention and treatment. Studies of the neurobiology of substance use and psychiatric disorders are accumulating rapidly and informing treatment development. Information about the prevention and treatment of infectious diseases and other medical conditions associated with substance use disorders also is growing, and it is important that patients are able to benefit from this. The articles in this issue provide state-of-the-art information about several issues related to comorbidity in substance use disorders. PMID- 15550283 TI - Forensic and medical legal issues in addiction psychiatry. AB - There is a broad range of forensic issues in addiction psychiatry. For many psychiatrists, what is most challenging about addressing these issues is their origin in the law, which is a system and a way of thinking that may feel foreign to the medical practitioner. To address forensic issues, addiction psychiatrists should learn and understand the specific legal questions that arise in each forensic context. They should become familiar with the relevant definitions, criteria, and legal requirements that apply in each specific area of their practice, rather than assume that clinical definitions and reasoning will carry them. If they perform forensic evaluations, addiction psychiatrists must distance themselves from the wish to help the examinee, focusing on the role of neutral examiner. Comfort and effectiveness with forensic issues require familiarity with, knowledge of, and ultimately respect for the forensic contexts of addiction psychiatric practice. PMID- 15550284 TI - Pharmacologic management of relapse prevention in addictive disorders. AB - Substance use disorders are an important public health problem associated with significant mortality and morbidity. Effective maintenance pharmacotherapies are available for tobacco, alcohol, and opioid use disorders. For optimum treatment response, these medications should be used in conjunction with behavioral interventions. For other drugs of abuse, especially for cocaine, medication development is an active area of research. Further research is needed to develop new pharmacotherapies for substance use disorders and establish clinical guidelines on how to use these medications most effectively. PMID- 15550285 TI - Diagnosis and treatment of co-occurring affective disorders and substance use disorders. AB - There is growing interest in the co-occurrence of mood and substance use disorders. It is clear that co-occurrence of these disorders is common and has an impact on prognosis and course of both disorders. The diagnostic issues at the interface of substance or alcohol use disorders and affective illnesses are particularly difficult because of the substantial symptom overlap between substance intoxication and withdrawal and symptoms of affective disorders. Over the past few years, advances have been made in the treatment of co-occurring disorders. Further investigation of specifically tailored treatments for patients with co-occurring substance use and other mood disorders is underway. Because many advances have been made in pharmacotherapy of mood disorders in the past 10 years, this progress will impact individuals with co-occurring disorders, because newer agents with less toxicity and fewer adverse effects and interactions with substances of abuse will be evaluated for treating the comorbid condition. Specific considerations in choosing a pharmacologic agent for use in patients with substance use disorders include safety, toxicity, and abuse liability. Although there are few studies specifically targeting pharmacotherapy for co occurring disorders, those that have been conducted indicate that similar pharmacotherapeutic agents work for mood disorders with or without substance use disorders. In conclusion, although the co-occurrence of substance abuse and mood disorders is an important area in which recent developments provide cause for considerable optimism, much work remains to be done. PMID- 15550286 TI - Neurobiological mechanisms in addictive and psychiatric disorders. AB - The studies reviewed indicate that brain stress system play an important role in the acquisition and maintenance of drugs of abuse that target the brain's reward centers. In doing so, they may destabilize these areas, making the perception of pleasure more elusive and difficult to attain. Withdrawal from drugs of abuse leads to the activation of brain CRF systems that may produce the anxiogenic response associated with drug withdrawal. More research, however, is needed to investigate the role of brain stress systems and neuropeptides in other drug withdrawal symptoms such as anhedonia. A better understanding of the brain systems underlying drug withdrawal may help in the development of improved pharmacotherapies that can alleviate drug withdrawal symptoms. The second part of the article indicated that there is a very high comorbidity between depression and drug dependence. The reviewed studies suggest that depressed patients initiate drug-taking behavior to self-medicate the symptoms associated with their psychiatric disorder. Chronic use of drugs of abuse, however, may exacerbate the symptoms of pre-existing mental disorders and subsequently increase drug-taking behavior. Conversely, professional treatment of pre-existing psychiatric disorders may decrease the use of illicit substances. PMID- 15550287 TI - Medical comorbidity in addictive disorders. AB - Greater risks exist for medical comorbidities in persons with addictive disorders. Clinicians should screen for early comorbidities such as hepatitis C and HIV. During acute intoxications and overdoses, patients are at greater risk for major respiratory and cardiac events. Long-term management of persons with addictions and medical disorders requires integrative care programs involving comprehensive primary and psychiatric care. PMID- 15550288 TI - Prescription opiate medications: medical uses and consequences, laws and controls. AB - The proposed analysis and evaluation of the data elements in the OPP and other similar regulatory programs will support the following potential impact on the patients and physicians in Michigan and other states: Reduced rates of addictive use of prescriptions of Schedule II medications. Reduced rates of addictive patterns of prescribing of Schedule II medications. Improved the prescribing of Schedule II medication for pain disorders. Improved the prescribing of Schedule II medications in addictive disorders. Establish the need and direction for development of curriculum for Schedule II drugs for undergraduate medical education and continuing medical education. Establish the need and direction for development of curriculum for use of Schedule II medications in patients with addictive and pain disorders. Explore the need and direction for development of the monitoring system medical curriculum for Schedule III, IV, and V drugs. Demonstrate link between diversion and adverse effects on health caused by an addictive pattern of use and prescribing of Schedule II drugs PMID- 15550289 TI - Behavioral interventions for dual-diagnosis patients. AB - Dual diagnosis patients come to treatment with a variety of deficits,talents, and motivations. A biopsychosocial treatment plan involves multiple interventions, including medications, medical treatment, psychotherapy, family therapy, housing, and vocational rehabilitation. Treatment must be individualized and integrated, and this requires collaboration among a variety of health caregivers. There is empirical evidence that dual-diagnosis patients can be helped to stabilize, to remain in the community,and even to enter the workforce. Behavioral interventions are key ingredients to integrated and comprehensive treatment planning. There is no single model for dual disorders that explains why substance use and psychiatric illness co-occur so frequently. Mueser et al described four theoretical models accounting for the increased rates of comorbidity between psychiatric disorders and substance use disorders. They suggested that there could be a common factor that accounts for both, primary psychiatric disorder causing secondary substance abuse, primary substance abuse causing secondary psychiatric disorder, or a bidirectional problem, where each contributes to the other. There is evidence for each, although some are more compelling than others, and none is so compelling that it stands alone. Although family studies and genetic research could explain the common factor, no common gene has appeared. Antisocial personality disorder has been associated with very high rates of substance use disorders and mental illness; however, its prevalence is too low to explain most of the co-occurring phenomena. Common neurobiology, specifically the dopamine-releasing neurons in the mesolimbic system, also may be involved in mental illness, but this is not compelling at the moment. The Self-medication model is very appealing to mental health professionals, as an explanation for the secondary substance abuse model. Mueser et al suggest that three lines of evidence would be present to support this explanation: (1) patients would report beneficial effects of substance use on their symptoms; (2) epidemiology would report that a specific substance would be used by specific psychiatric disorders, and (3) psychiatric patients with severe symptoms would be more likely to abuse substances than those with mild symptoms. Unfortunately the research data do not support these. The primary substance abuse causing secondary psychiatric disorder model could be explained by neuronal kindling from substance-induced disorders. Patients who develop the psychiatric disorder after the substance use disorder do have a course of illness similar to those with a psychiatric disorder, but without substance use disorder. The bidirectional model is consistent with the tendency of disturbed teenagers to socialize with youth using alcohol and drugs; however, this model has not been tested rigorously in research studies. With such a disparate set of models, behavior interventions are conceptualized best as a multi-component program, a treatment plan that generates a problem list and devises an intervention to respond to each member of the list. This requires a talented, multi-disciplinary team or network that can assess carefully and package the interventions creatively, and dose the treatment components empathically to fit the patient's tolerance, motivation, and abilities. PMID- 15550290 TI - Changing the world: the design and implementation of comprehensive continuous integrated systems of care for individuals with co-occurring disorders. AB - This article has described the CCISC model and the process of implementation of systemic implementation of co-occurring disorder services enhancements within the context of existing resources. Four projects were described as illustrations of current implementation activities. Clearly, there is need for improved services for these individuals, and increasing recognition of the need for systemic change models that are effective and efficient. The CCISC model has been recognized by SAMHSA as a consensus best practice for system design, and initial efforts at implementation appear to be promising. The existing toolkit may permit a more formal process of data-driven evaluation of system, program, clinician, and client outcomes, to better measure the effectiveness of this approach. Some projects have begun such formal evaluation processes, but more work is needed, not only with individual projects, but also to develop opportunities for multi system evaluation, as more projects come on line. PMID- 15550291 TI - Occupational exposure and addictions for physicians: case studies and theoretical implications. AB - Risk of addiction through occupational exposure to drugs of abuse is an important but relatively neglected public health problem. Stress and access may have much less of a role in addiction among certain populations than originally was thought. Risk of addition may be increased dramatically by unintentional exposure in the workplace to potent substances that sensitize the brain. Everyone knows that second-hand inhalation of crack vapors is a very dangerous proposition, but rarely has alarm been raised about exposing anesthesiologists to second-hand fentanyl. Additional studies of the relationship between exposure in the workplace and addiction are necessary. These studies should include biological measures, such as blood levels in exposed workers, and sensitive assays that quantitatively assess levels of exposure in the workplace. PMID- 15550292 TI - The basal ganglia: anatomy, physiology, and pharmacology. AB - The basal ganglia are perceived as important nodes in cortico-subcortical networks involved in the transfer, convergence, and processing of information in motor, cognitive, and limbic domains. How this integration might occur remains a matter of some debate, particularly given the consistent finding in anatomic and physiologic studies of functional segregation in cortico-subcortical loops. More recent theories, however, have raised the notion that modality-specific information might be integrated not spatially, but rather temporally, by coincident processing in discrete neuronal populations. Basal ganglia neurotransmitters, given their diverse roles in motor performance, learning, working memory, and reward-related activity are also likely to play an important role in the integration of cerebral activity. Further work will elucidate this to a greater extent, but for now, it is clear that the basal ganglia form an important nexus in the binding of cognitive, limbic, and motor information into thought and action. PMID- 15550293 TI - The neuropsychiatry of Parkinson's disease and related disorders. AB - Parkinson's disease is associated with classical Parkinsonian features that respond to dopaminergic therapy. Neuropsychiatric sequelae include dementia, major depression, dysthymia, anxiety disorders, sleep disorders, and sexual disorders. Panic attacks are particularly common. With treatment, visual hallucinations, paranoid delusions, mania, or delirium may evolve. Psychosis is a key factor in nursing home placement, and depression is the most significant predictor of quality of life. Clozapine may be the safest treatment for psychotic features, but more research is needed to establish the efficacy of antidepressant treatments. Dementia with Lewy bodies, the second most common dementia in the elderly, may present in association with systematized delusions, depression, or RBD. Early evidence suggests the utility of rivastigmine, donepezil, low-dose olanzapine, and quetiapine in treating DLB. Parkinson-plus syndromes generally lack a good response to dopaminergic treatment and evidence additional features, including dysautonomia, cerebellar and pontine features, eye signs, and other movement disorders. MSA is associated with dysautonomia and RBD. SND (MSA-P) is associated with frontal cognitive impairments, but dementia, psychosis, and mood disorders have not been strikingly apparent unless additional pathological findings are present. In SDS (MSA-A), impotence is almost ubiquitous; urinary incontinence is frequent; depression is occasional, and sleep apnea should be treated to avoid sudden death during sleep. OPCA neuropsychiatric correlates await further definition. Progressive supranuclear palsy neuropsychiatric features include apathy, subcortical dementia, pathological emotionality, mild depression and anxiety, and lack of appreciable response to donepezil. CBD usually is recognized by early frontal dementia with ideomotor apraxia, often in the right upper extremity, attended later by poorly responsive unilateral Parkinsonism, with additional signs including cortical reflex myoclonus, limb dystonia, alien limb, oculomotor apraxia when asked to look horizontally, depression, personality changes, and, occasionally, Kluver-Bucy syndrome. The neuropsychiatry of FTDP-17 involves apraxia, executive impairment, personality changes, hyperorality, and occasional psychosis. Future research in these Parkinsonian disorders should target the characterization of neuropsychiatric sequelae and their treatment. PMID- 15550294 TI - Some mechanisms linking economic inequality and infant mortality. AB - We use data from the 1985, 1987 and 1991 United States Vital Statistics Linked Infant Birth and Death Records to assess the relationship between state-level economic inequality and an infant's probability of death. We find that economic inequality is associated with higher neonatal mortality even after we control mother's age and race and state characteristics that are likely to be associated with both inequality and infant death. Inequality is not associated with post neonatal mortality. We assess three mechanisms that could link income inequality and infant deaths: non-linearity in the relationship between parental income and infant death, economic segregation, and state health care spending. Our evidence suggests that non-linearity in the relationship between family income and infant health accounts for little of the relationship between inequality and infant death. However inequality is associated with greater economic segregation, which in turn is associated with a higher probability of infant death. This effect is partially offset by the fact that inequality is also associated with state spending on health care, which is in turn associated with lower death rates. The increase in economic segregation increased infant deaths more than the increase in health care spending reduces them, so the net effect of economic inequality is to increase infant deaths especially in the first month after birth. PMID- 15550295 TI - Determinants of old-age mortality in Taiwan. AB - Relationships among socio-demographic characteristics, general assessments of health, and old-age mortality have been well established in developed countries. There is also increasing focus on the connection between early-life experiences and late-life health. This study tests these and other associations using representative survey data from Taiwan. Survey data on the 60 and over population (N = 4049) in 1989, 1993, and 1996 are linked to 1989-1999 data from a national death registry. Survival is modeled using Gompertz regression and both fixed and time-varying covariates. Possible effects of Taiwan's Universal Health Insurance Program are tested by coding observations as pre- or post-insurance. Some results replicate findings from developed countries; others are new. Contemporaneous education effects on survival are attenuated after introducing some indicators of health from a previous period. Functional and global assessments of health have stronger associations with survival than reports of other health-related characteristics. Mainlanders have higher survival than others. Increases in survival over the 1990s were concentrated among those with the greatest number and severity of functional limitations. This pattern may be an indication of better access to health care or changes in longer-run, but unmeasured, determinants of old-age mortality. PMID- 15550296 TI - Causes and demographic, medical, lifestyle and psychosocial predictors of premature mortality: the CARDIA study. AB - We examined the 16-year mortality experience among participants in the baseline examination (1985-86) of the Coronary Artery Risk Development in Young Adults (CARDIA) Study, a U.S. cohort of 5115 urban adults initially 18-30 years old and balanced by sex and race (black and whites) in the USA. We observed 127 deaths (annual mortality of 0.15%). Compared to white women, the rate ratio (95% confidence interval) of all-cause mortality was 9.3 (4.4, 19.4) among black men, 5.3 (2.5, 11.4) among white men and 2.7 (1.2, 6.1) among black women. The predominant causes of death, which also differed greatly by sex-race, were AIDS (28% of deaths), homicide (16%), unintentional injury (10%), suicide (7%), cancer (7%) and coronary disease (7%). The significant baseline predictors of all-cause mortality in multivariate analysis were male sex, black race, diabetes, self reported liver and kidney disease, current cigarette smoking and low social support. Two other factors, self-reported thyroid disease and high hostility, were significant predictors in analyses adjusted for age, sex and race. In conclusion, we found striking differences in the rates and underlying cause of death across sex-race groups and several independent predictors of young adult mortality that have major implications for preventive medicine and social policies. PMID- 15550297 TI - Smoking, respiratory disease and health service utilisation: the paradox. AB - We compared need for and utilisation of health services among young, healthy smokers who did not want to quit "the non-motivated", and "motivated smokers", relative to never-smokers. This cross-sectional study included 9915 Hong Kong police officers. We assessed smoking status, differentiating non-motivated from motivated smokers. Need was measured as the number of respiratory symptoms, and utilisation as the number of doctor consultations. All smokers had significantly more respiratory symptoms, but relatively lower utilisation. After adjusting for other factors, the odds ratio for utilisation was 0.77 (95% CI 0.64-0.93) for motivated smokers and 0.62 (95% CI 0.50-0.77) for non-motivated, relative to never-smokers (p for trend < 0.001). Young, relatively healthy smokers, particularly the non-motivated, utilise less health services relative to the amount of symptoms. This has implications for planning smoking cessation services. PMID- 15550298 TI - Exploring the association of John Henry active coping and education on smoking behavior and nicotine dependence among Blacks in the USA. AB - Although smoking is used as a coping tool in response to stress and Blacks have been found to report smoking more in response to stress than Whites, little research exists that has examined ethno-culturally specific constructs of stress and coping as they relate to smoking behavior and nicotine dependence among Blacks in the USA. This study explored the association between the ethno culturally interactively defined construct of John Henryism, as well as the individual contributions of John Henry active coping and education on smoking behavior and nicotine dependence in a relatively urban-Midwestern Black population. Self-identified Black patients (n = 146) who had previously received a clinical intervention for nicotine dependence were followed to assess smoking status and John Henry active coping. Results revealed that patients with low levels of education who had low levels of John Henry active coping reported higher nicotine dependence scores than any other education by John Henry active coping group. Furthermore, low levels of John Henry active coping were associated with the use of menthol cigarettes and lower-educational level was associated with smoking greater than 20 cigarettes per day. Further community-based studies examining this construct among Black smokers in various socio-cultural contexts are needed to clarify the association between John Henry active coping and socioeconomic status on smoking behavior and nicotine dependence among Blacks. PMID- 15550299 TI - Individual and district scale determinants of users' satisfaction with primary health care in developing countries. AB - The paper has two aims: to explore possible determinants of user satisfaction based on a broad conception of the health care system involving both individual and district scale variables, and to add to the limited knowledge and empirical study about such determinants in developing countries. The study was carried out in one of Brazil's poorest states, Ceara, in the northeast of the country. Brazil has a policy of an increased role for the population in management together with decentralised management to the local administrative (municipio) scale. For user views on health care to be useful in management, understanding their determinants needs to explore not only individual scale factors but also influences from the local health system and wider society, here termed district scale factors. The study design took different angles on users' satisfaction to build three measures: the particular health event (termed perceived quality); a generalised perception (termed satisfaction); and, satisfaction with the Community Health Worker (termed CHW satisfaction). Potential determinants explored at the individual scale (17 variables) are grouped into: socio-demographic and economic characteristics; health outcome; health care provision; and, awareness of space for local voice in planning. Potential determinants explored at the district scale (50 variables) are grouped into: formal organisation and management (health system inputs, management capacity, outputs and outcomes); informal organisation (population awareness of health system activities, staff characteristics, district management style); and, local political culture (geographic and socio economic population profile, commitment to the district of local leaders, norms and values of staff regarding practice). Three determinants were key for all three user evaluation measures: getting an appointment, getting better, and the type of district (rural-urban). Our primary conclusion from this study is that there are limitations to the extent that user satisfaction can fulfill the claims made for it. On a more positive note, with these limitations noted, user satisfaction can prove a useful management tool more modestly at a local, context specific scale. PMID- 15550300 TI - Quantifying the effect of health status on health care utilization using a preference-based health measure. AB - The purpose of this study was to quantify the effect of health status on current and future payments and number of visits to health professionals in a large, representative community sample in British Columbia, Canada. The study population was comprised of all respondents to the 1994/5 cycle of the Canadian National Population Health Survey (NPHS) who were 12 years of age or older and residing in the province of British Columbia (N = 2084). Health status was measured with the Health Utilities Index (HUI). Two outcomes were defined for each subject: (a) the sum of all healthcare costs covered by the Medical Services Plan, incurred during a given fiscal year, and (b) the total number of visits to all health practitioners during the same year. Outcome data were obtained for a period 1994 1998. We examined the relationship between the HUI and healthcare use in a multivariate log-linear model. In the full sample, better health in 1994-1995 was associated with lower healthcare cost and lower number of visits from 1994 through 1998. The overall adjusted cost ratio was 0.89 (99% CI = 0.85, 0.94) and the overall adjusted visit ratio was 0.91 (99% CI + 0.87, 0.95). The effect of health status on the costs of care and on the number of visits was similar in men and women, was stronger in persons less than 45 years of age compared to those 45+, and was not different according to place of residence. We conclude that the HUI is a strong predictor of health services use over 5 years. A 0.1 improvement in health utility is associated with a 10% reduction in the costs of care and number of visits to health professionals. PMID- 15550301 TI - Do divergences between stated and actual willingness to pay signify the existence of bias in contingent valuation surveys? AB - This study's objective is to determine the factors that cause divergences over time (differences) between stated willingness to pay (WTP) and actual WTP (purchase behaviour), and consider whether any divergence signifies the presence of bias in contingent valuation studies. Stated WTP for insecticide-treated bed nets (ITNs) was elicited from a random sample of respondents using three question formats in Nigeria. The question formats were the bidding game (BG), binary with follow-up (BWFU) and a novel structured haggling (SH) technique. The sales of the nets and a second survey were conducted 1 month after the first survey. In the second survey, factors that might explain the divergences were built into the questionnaire and these together with socio-economic variables were examined for causes of divergences in WTP. Data were analysed using non-parametric tests, testing of means and cross-tabulations. There were divergences in WTP in all three question formats: 69.4% in the BG, 78.7% in the BWFU and 48.8% in the SH. The higher the stated WTP, the more likely the divergence between stated and actual WTP. The attitude of the community leaders to the ITNs in the BG (p<0.05), the time respondents had to think about their WTP (p<0.05) and the external information they received about the ITNs in the BWFU (p<0.05) all led to divergences in WTP. We conclude that there are genuine causes of divergences between stated and actual WTP across the three question formats, and that the lesser the criterion validity score, the more the level of divergence in WTP. Studies that compare stated and actual WTP should explicitly determine the causes of divergences in order to assess the role of bias in the divergences. PMID- 15550302 TI - Economic antecedents of the Swedish sex ratio. AB - Literature describing environmental influences on human conception and gestation implies that the ratio of male to female live births should vary positively over time with the population's ability to produce and distribute goods and services. No direct test of this hypothesis appears in the literature despite its apparent importance in understanding the biological implications of collective choices. We offer a test based on Swedish data for the years 1862 through 1991. The results support the hypothesis. We argue that the findings have implications for basic science as well as for the debate over the biological effects of social policy. PMID- 15550303 TI - Evidence-based medicine and its implications for the profession of chiropractic. AB - Evidence-based medicine (EBM) has grown in popularity and prominence in the world of orthodox medicine since the 1980s. The focus of this article is on the process of developing practice guidelines (one type of EBM) and its effects upon chiropractic, a profession with a "philosophy, science and art" that is constructed upon divergent epistemological and methodological tenets (namely, the idea of "vitalism"). The EBM movement is conceptualized as part of a larger political economy surrounding the health care environment that creates a new set of imperatives for orthodox medicine, and also branches of alternative medicine that are in the process of professionalization. The quantitative, positivist and empiricist assumptions of EBM dictate which approaches to treatment and which clinical procedures are legitimate and perhaps reimbursable under systems of managed care. The ramifications of practice guidelines and its effects upon the intraprofessional segments of the chiropractic profession are also discussed. PMID- 15550304 TI - Aggressive active case detection: a malaria control strategy based on the Brazilian model. AB - Since 1996, the Brazilian Ministry of Health has adopted a malaria control strategy known as aggressive active case detection (AACD) in which most or all members of every community are tested and treated for malaria on a monthly basis. The strategy attempts to identify and treat cases of asymptomatic malaria, which, if untreated, continue to transmit the infection. Malaria remains uncontrolled because almost all health care systems in the world rely on passive case detection: the treatment of only symptomatic cases of malaria. Research has shown conclusively that asymptomatic cases exist in any population where malaria transmission is stable and incidence is high: therefore passive case detection simply will not succeed in breaking the cycle of transmission. Numerous case studies show that malaria has been successfully controlled on a regional or national level by mass blood surveys. AACD is an effective malaria control strategy if used in conjunction with other methods, especially when (1) an effective treatment exists, (2) influx of potential carriers of the infection can be monitored, and (3) people are inclined to cooperate with monthly blood testing. AACD requires access to rapid diagnostic tests (RDTs), microscopy supplies, extensive human resources, and prompt, affordable, and effective treatment. AACD is compared to PCD in terms of clinical efficacy and cost effectiveness in a case study of malaria in the Brazilian Yanomami Indians. Where it is feasible, AACD could drastically reduce the incidence of malaria and should be an integral part of the World Health Organization's Roll Back Malaria strategy. PMID- 15550305 TI - A tailored intervention to promote breast cancer screening among South Asian immigrant women. AB - This study developed and evaluated a socioculturally tailored intervention to improve knowledge, beliefs and clinical breast examination (CBE) among South Asian (SA) immigrant women. The intervention comprised a series of socioculturally tailored breast-health articles published in Urdu and Hindi community newspapers. A pre- and post-intervention design evaluated the impact of the mailed articles among 74 participants. The mean age of participants was 37 years (SD 9.7) and they had lived 6 years (SD 6.6) in Canada. After the intervention, there was a significant increase in self-reporting 'ever had' routine physical checkup (46.4-70.8%; p < 0.01) and CBE (33.3-59.7%; p < 0.001). Also, the total summed scores of accurate answers to 12 knowledge items increased (3.3-7.0; p < 0.001). For constructs of health belief model, participants rated their level of agreement for a number of items on a scale of 1-4 (disagree to agree). After the intervention the following decreased: misperception of low susceptibility to breast cancer among SA immigrant women (3.0-2.4; p < 0.001); misperception of short survival after diagnosis (2.7-1.8; p < 0.001); and perceived barriers to CBE (2.5-2.1; p < 0.001). Self-efficacy to have CBE increased (3.1-3.6; p < 0.001). The change scores of five predictor variables were entered in a direct logistic regression to predict the uptake of CBE among participants who never had it prior to the intervention. The model, as a set, was statistically reliable [chi2(5, n = 48) = 14.2 , p < 0.01] and explained 35% of variance in the outcome; perceived barriers remained an independently significant predictor. The results support the effectiveness of written socioculturally tailored language-specific health education materials in promoting breast cancer screening within the targeted population. Future research should test the intervention in other vulnerable populations. PMID- 15550306 TI - Between 'desperation' and disability rights: a narrative analysis of complementary/alternative medicine use by parents for children with Down syndrome. AB - This paper presents a narrative analysis of complementary/alternative medicine (CAM) use by parents for children with Down syndrome (DS), based on interviews conducted with thirty families. Critics often presume that CAM use for children with developmental disabilities reflects parental desperation in the face of limited biomedical options. Integrating insights from anthropological studies of CAM with narrative analyses in disability studies, we constructively complicate this interpretation in two ways. First, we suggest that the appeal of CAM may lie in its discursive consonance with the broader narrative strategies through which parents construct alternatives to conventional definitions of DS as a condition with a fixed, universal, and essentially pathological course. Second, we submit that the process of seeking and evaluating information about CAM is consonant with how parents construct their identities as 'good' parents through describing their roles as committed advocates and service coordinators for their children. In these ways, CAM can be conceptualized as a new discursive resource that parents engage in their culturally and historically specific efforts to articulate the essential human rights of their children, and to assert the moral soundness of their own parenthood. These findings provide a new conceptualization of parents' motives for choosing CAM, thereby posing new questions for further research about CAM use for developmental disabilities. PMID- 15550307 TI - Influence of social support on progression of coronary artery disease in women. AB - There is strong and consistent evidence across numerous studies that social isolation or lack of social support is an independent risk factor for incident coronary heart disease. However, the impact of social isolation or lack of social support on the progression of coronary atherosclerosis in women has not been well documented. Among 292 women, aged 30-65 years, consecutively hospitalized with acute myocardial infarction or unstable angina between 1991 and 1994 enrolled in the female coronary risk study, 102 women were available to study disease progression over an average of 3.2 years. Three aspects of social support were studied: emotional support, social integration, and interpersonal social relations. Quantitative coronary angiography was performed 3-6 months following index hospitalization and repeated 3 years later. Progression of coronary atherosclerosis was evaluated as the change in mean luminal diameter from first to second measurements of 10 pre-defined coronary segments. Mixed model ANOVA was used to analyze the impact of social support on progression of coronary atherosclerosis. Significantly greater coronary atherosclerosis progression was found among women who lacked emotional support (mean coronary artery luminal diameter narrowing by 0.15 mm), with social isolation (0.14 mm), and lack of interpersonal social relations (0.13 mm), whereas women with high levels of support progressed less. It was 0.05 mm in women with high levels of emotional support, 0.07 mm for socially integrated women, and 0.04 mm in women with adequate interpersonal social relations. These associations were independent of conventional clinical and lifestyle factors such as age, smoking history, body mass index, menopausal status, and diagnosis of index event of acute myocardial infarction. The results of our study suggest that lack of emotional support, social isolation, and lack of interpersonal social relations are important risk factors for accelerated progression of coronary atherosclerosis in middle-aged women. PMID- 15550308 TI - Shifting the demand for emergency care in Cuba's health system. AB - Cuba has developed a programme of quality improvement of its health services, which includes an extramural emergency care system in which polyclinics and general practitioner networks play an important role. Using routine health information from the decentralised first line emergency units (FLES) and from the hospital emergency service (HES) for the period 1995-2000, we evaluated the effects of the emergency care subsystem reform on the utilisation rates of first line and hospital services in Baracoa and Cerro, a rural and a metropolitan municipality, respectively. In the self-contained health system of Baracoa, the reform of the emergency subsystem resulted in a first phase of increased utilisation of the FLES, followed by a second phase of gradual decrease, during which there was an increased utilisation of general practitioners. In contrast, the overall results of the reform in Cerro were unclear. The proximity to a hospital seems to be the most important element in the patient's decision on which entry point to the Cerro health system to use. A potential adverse effect of the reform is an increased emergency services utilisation in situations where GP care remains below patients' expectations. Given the current world-wide trends in health-care reform, the organisational alternatives developed in the Cuban health system might remain specific to the local contextual setting. PMID- 15550309 TI - Trend and area variation in amphetamine prescription usage among children and adolescents in Michigan. AB - The increased use of stimulant medications for children and teenagers is an ongoing issue of professional and public concern. Unlike methylphenidate, the growth of prescriptions and patterns of utilization of amphetamines for pediatric populations have not been well documented. The study objectives were to describe the trends of amphetamine prescription utilization among pediatric age groups in Michigan and to compare area variations. A population-based computerized data set from the state of Michigan was used to extract all outpatient prescriptions for Schedule II amphetamines dispensed from 1990 to 1997. The prescribing rates by age groups and by counties were computed with the projected population size of corresponding years, and mapped and analyzed with spatial statistical methods. Counties that did not conform to the global spatial dependence pattern in the prescription rate were identified using Moran scatter plot. A total of 236,661 outpatient prescriptions for amphetamines were dispensed in Michigan during the time frame, including less than 1% for methamphetamine, 24.5% for amphetamine, and 74.8% for dextroamphetamine. The prescribing rate was highest among children 10-14 years old (380 prescriptions per 10,000 people) in 1997, followed by children 5-9 years old (253 prescriptions per 10,000). Over the 8-year period, the prescribing rates of amphetamines increased significantly, ranging from 380% for children 2-4 years to 817% for teenagers older than 14 years. The rates among counties ranged from 60 to 1648 per 10,000 children 5-14 years old, with the highest prescribing rates in the northwestern regions of Michigan. Although spatial dependence explained 36% of the variance, the area variations that are unaccounted for are still considerable. In conclusion, there were substantial increases and unexplained area variations in amphetamine usage in Michigan during the study period. Both phenomena require awareness and evaluation from researchers, policy makers, clinicians, and the public. PMID- 15550310 TI - Injury recurrence among untreated and medically treated victims of violence in the USA. AB - Studies on the risk of intentional re-injury tend to use samples of admissions to hospitals and conclude that prior intentional injury is predictive of future such admissions. Limiting samples to persons receiving medical attention misses the equally important population of those intentionally injured but not receiving medical attention. Approximately 2 million violence-related injuries go medically unevaluated each year in the United States. A significant portion of these are severe. Including this population captures the dark figure of untreated intentional injury and allows for the estimation of the effects of medical intervention on future risk. This paper tests the hypothesis that those going without medical evaluation will have higher odds of injury recurrence than the traditionally studied population of those receiving health care. Data are drawn from the National Crime Victimization Survey (NCVS), an ongoing and nationally representative longitudinal survey of households in the United States. The NCVS collects information at the household and person-level, and when victimization is identified through screener questions, at the incident-level. In addition to capturing crime and violence reported to the police and medical professionals, this methodology captures incidents that go unreported and untreated. Controlling for correlates of violence and reinjury, persons injured in violent events receiving medical attention alone, or in combination with police notification, are at substantially lower risk of incurring future violence and violence-related injuries than persons injured but not receiving medical or police attention. PMID- 15550311 TI - Widowhood and depressive symptoms among older Chinese: do gender and source of support make a difference? AB - In this study, we examined the effects of gender and pre-bereavement social support from three different sources (spouse, adult children, and friends) on widowhood adjustment among older adults in China. Hypotheses were developed by integrating the literature in the West and the cultural context of China. Data came from a panel survey, conducted in 1991 (baseline) and 1994 (follow-up), of a probability sample of older persons in Wuhan, China. For the present analysis, only those who were married with children at baseline were selected (N = 1,263). About 10% of the sample experienced spousal death between the two measurement points. Multiple regression analyses suggest that widowhood had a negative mental health consequence for older Chinese. Social support from adult children buffered the deleterious effect of widowhood, whereas spousal support during the marriage increased one's vulnerability. Support from friends was not found to have a significant effect. Gender difference in the effect of widowhood was also not evident. In this study, we have extended bereavement and social support research to a developing nation, with some findings similar to and some different from studies in Western developed nations. PMID- 15550312 TI - Perceptions of risk and strategies for prevention: responses to HIV/AIDS in rural Malawi. AB - This paper combines quantitative and qualitative data to investigate changes in perceived risk of contracting HIV/AIDS in rural Malawi. Using longitudinal survey data, we find that Malawians worried less about contracting HIV/AIDS in 2001 than in 1998. According to qualitative interviews and observational journal accounts, HIV/AIDS and strategies to prevent it are a frequent topic of conversation amongst married Malawians. Women report worrying most about their husbands as a possible source of infection, discussing with them the importance of avoiding infection, and, increasingly, using divorce to reduce their risk. Men report worrying most about their extramarital partners and adopting preventive strategies such as fewer partners and more careful partner selection. We show that the decline in perceived risk is significantly associated with declines in the behaviors that Malawians worry most about and perceptions of risk in individuals' social networks. We interpret these findings as evidence that Malawians are changing their behavior in ways that may reduce the spread of HIV/AIDS. PMID- 15550313 TI - High prevalence of hepatic focal nodular hyperplasia in subjects with hereditary hemorrhagic telangiectasia. AB - A vascular pathogenesis of hepatic focal nodular hyperplasia (FNH) has been suggested; this study was aimed to evaluate in families with hereditary hemorrhagic telangiectasia (HHT) the prevalence of FNH, relating it to presence and stage of hepatic vascular malformations (VMs). Fifty-two HHT families underwent a screening program including abdominal Doppler sonography (US) searching for hepatic VMs; we classified them as minimal, moderate and severe, depending on the number and degree of abnormalities found by Doppler US. Presence of focal liver lesions was recorded. Diagnosis of FNH was made if at least two examinations, whether color Doppler US, liver scintigraphy, dynamic computed tomography (CT) or magnetic resonance (MR), showed suggestive findings. FNH was found in five out of 274 subjects (1.8%). All five were affected by HHT. Thus, percentage related to the group of affected patients increased to 2.9; 4/5 presented severe liver VMs. Female-to-male ratio was 4:1. FNH was single in three cases; tumor size ranged between 20 and 90 mm. During follow-up, no lesion showed a reduction in size, three showed an increase. Prevalence of FNH in patients with HHT is far greater than that reported in the general population; Doppler US role in its diagnosis and follow-up is highlighted. PMID- 15550314 TI - Compliance of abdominal aortic aneurysms: evaluation of tissue Doppler imaging. AB - Expansion of abdominal aortic aneurysms (AAA) is due to remodeling of the parietal extra-cellular matrix and may lead to rupture. This remodeling is reflected by compliance which may be an indicator of AAA behavior and thus useful for clinicians. Tissue Doppler Imaging (TDI) is an ultrasonographic modality which allows wall motion measurements along an arterial segment. It has previously been evaluated in normal aortas and was then evaluated in thirty-five patients with AAA. Mean values (+/-standard deviation) characterizing maximum diameter AAA compliance were dilation 809 mum (+/-465), strain 2.2% (+/-1), pressure strain elastic modulus 3.94 10(5) Pa [3.25; 4.8] and stiffness 28.8 [24; 34.5], last values being expressed as geometric mean [interquartile range]. They were in accordance with those previously obtained with other systems. Segmental parameter values were maximum mean segmental dilation 534 mum (+/-305) and segmental compliance 14.6 (+/-8.3) 10(-2) mum/Pa. Reproducibility was appropriate for clinical studies. The TDI system is simple and reliable for measurement of AAA compliance, and compliance can easily be recorded during routine ultrasound control. PMID- 15550315 TI - Volume estimation of small phantoms and rat kidneys using three-dimensional ultrasonography and a position sensor. AB - To evaluate the accuracy of small volume estimation, both in vivo and in vitro, measurements with a three-dimensional (3D) ultrasound (US) system were carried out. A position sensor was used and the transmitting frequency was 10 MHz. Balloons with known volumes were scanned while rat kidneys were scanned in vivo and in vitro. The Archimedes' principle was used to estimate the true volume. For balloons, the 3D US system gave very good agreement with true volumes in the volume range 0.1 to 10.0 mL (r = 0.999, n = 45, mean difference +/- 2SD = 0.245 +/- 0.370 mL). For rat kidneys in vivo (volume range 0.6 to 2.7 mL) the method was less accurate (r = 0.800, n = 10, mean difference +/- 2SD = -0.288 +/- 0.676 mL). For rat kidneys in vitro (volume range 0.3 to 2.7 mL) the results showed good agreement (r = 0.981, n = 23, mean difference +/- 2SD = 0.039 +/- 0.254 mL). For balloons, kidneys in vivo and in vitro, the mean percentage error was 9.3 +/- 4.8%, -17.1 +/- 17.4%, and 4.6 +/- 11.5%, respectively. This method can estimate the volume of small phantoms and rat kidneys and opens new possibilities for volume measurements of small objects and the study of organ function in small animals. (E-mail ). PMID- 15550316 TI - A method to create reference maps for evaluation of ultrasound images of carotid atherosclerotic plaque. AB - Ten formalin-fixed atherosclerotic carotid plaques removed by endarterectomy were molded into rectangular agar blocks containing fiducial markers on the top surface. Plaque and fiducial markers were imaged with 3-D multiangle ultrasound (US) spatial compounding as well as planar X ray. Subsequently, the blocks were decalcified, sliced, photographed and analyzed histologically. This gave a total of 123 slices. The plaque regions of the photographs were outlined and the outline adjusted to partly compensate for occasional displacement during slicing. Inside this outline, the material constitutions were found by incorporating the histologic information. From this set, slices with 1. too much tissue displacement due to cutting or 2. lack of identification of calcification as found by x ray, were removed. This resulted in 53 reference maps. The material types identified covered soft tissues, fibrous tissue, calcified tissue and unidentified tissues. The 53 reference maps can be used for direct automated quantitative comparison with US images. PMID- 15550317 TI - Parameters describing motion in carotid artery plaques from ultrasound examination: A reproducibility study. AB - We have previously developed a method for quantifying motion in carotid artery plaques from sequences of ultrasound (US) radiofrequency images. Here, we examine the intraoperator reproducibility of the results. Five independent recordings were made on each of six symptomatic and six asymptomatic patients, and processed off-line into 29 motion parameters, representing motion amplitude, stretch/compression and shear motion. For the statistical analysis, we used a linear mixed model and investigated the parameters for contributions from individual patients, contributions from recordings on each patient and contributions from heart cycles within each recording. The model was valid for seven parameters calculated over the entire heart cycle (four calculated over the systole only), which all showed good reproducibility (intraclass coefficient for variance over all patients rho(alpha) >/= 0.4). Averaging three recordings of two heart cycles each gives acceptable accuracy (normalised variance of patient means lambda < 0.3). This acquisition scheme is reasonable in a clinical situation. PMID- 15550318 TI - In vitro and in vivo studies on continuous echo-contrast application strategies using SonoVue in a newly developed rotating pump setup. AB - With emerging imaging strategies for contrast sonography (CS), there is a rising demand for the precise control of ultrasound (US) contrast agent delivery. Constant delivery minimizes artefacts and improves efficacy. The aim of this study was to evaluate the physical properties of the new contrast agent SonoVue and to evaluate the feasibility and accuracy of a new infusion approach using an automated infusion system for contrast agitation and delivery of echo-contrast agents. In vitro testing of infusion properties of SonoVue were performed in a capillary phantom mimicking tissue perfusion. Nonagitated standard infusion setups were compared with hand agitation and the new pump system with respect to possible artefacts, constancy of contrast effect and efficacy. In three volunteers, the new pump system was tested for constancy of contrast in large vessels. Without continuous agitation, continuous infusion of SonoVue resulted in bolus-like signal-intensity curves, along with substantial imaging artefacts. Additionally, homogenization of SonoVue significantly improved efficacy (p < 0.0001). No significant differences were found between hand agitation and homogenization by the new pump. In clinical settings, constant agitation using the new pump resulted in constant signal conditions in the carotid artery 3.72 +/ 0.46 units (U) after 5 min. Continuous agitation of SonoVue is mandatory for quantitative approaches. By the new infusion technique, CS could be performed for a reasonably long time period and efficacy is significantly improved (p < 0.0001). The new infusion technique might thereby allow routine application of constant infusion scenarios in clinical CS. PMID- 15550319 TI - Origins of the edge shadowing artefact in medical ultrasound imaging. AB - Ultrasound (US) B-mode images distal to smooth, rounded cavities, such as cysts, containing a fluid with a speed of sound mismatch to the surrounding tissue, often exhibit a "refractile" edge shadowing artefact. This usually appears as narrow, hypoechoic, shadow lines extending a significant distance distal to the lateral edges of the fluid cavity and parallel to the US beam. The true reasons for this artefact are likely to be complex and to vary from case to case, with many different explanations found in the literature. However, we present a simplified theoretical model for the phenomenon based on a pulsed, finite-beam solution of US scattering from circular fluid-filled cylinders that suggests that "edge" shadows can occur distal not only to edges but also to points where the incident beam intersects the cavity near to the critical angle. Both mechanisms support the view that edge shadows can arise from the combination of unusually high wavefront spreading and the speckle-generating nature of the surrounding tissue. In vitro data from a tissue-mimicking phantom confirms that the edge shadow structure depends on the sign of the speed of sound contrast between the cylinder fluid and the surrounding medium. PMID- 15550320 TI - Acoustic radiation force impulse imaging of the mechanical properties of arteries: in vivo and ex vivo results. AB - We present results of a pilot study of ex vivo and in vivo acoustic radiation force impulse (ARFI) imaging demonstrating measurements of the mechanical properties of the carotid and popliteal arteries. The results were obtained on a modified commercial scanner, providing coregistered B-mode and color Doppler images. 2D and 1D through time images are formed from the measurements of tissues' response to very brief and localized applications of radiation force. The images show good correlation with B-mode and, in ex vivo studies, pathology based characterizations of vessel geometry and plaque stiffness. In vivo measurements of arterial response during both systole and diastole are presented. We address implementation issues and discuss potential applications of this new vascular imaging method. PMID- 15550321 TI - Tissue displacements during acupuncture using ultrasound elastography techniques. AB - Acupuncture needle manipulation has been previously shown to result in measurable changes in connective tissue architecture in animal experiments. In this study, we used a novel in vivo ultrasound (US)-based technique to quantify tissue displacement during acupuncture manipulation in humans. B-scan ultrasonic imaging was performed on the thighs of 12 human subjects at different stages of needle motion, including varying amounts of rotation, downward and upward movement performed with a computer-controlled acupuncture needling instrument. Tissue displacements, estimated using cross-correlation techniques, provided successful mapping and quantitative analysis of spatial and temporal tissue behavior during acupuncture needle manipulation. Increasing amounts of rotation had a significant linear effect on tissue displacement during downward and upward needle motion, as well as on rebound tissue displacement after downward needle movement. In addition to being a valuable tool for studies of acupuncture's mechanism of action, this technique may have applications to other types of needling including biopsies. PMID- 15550322 TI - Theoretical derivation of SNR, CNR and spatial resolution for a local adaptive strain estimator for elastography. AB - Conventional techniques in elastography estimate the axial strain as the gradient of the displacement (time-delay) estimates obtained using cross-correlation of pre- and temporally stretched postcompression radiofrequency (RF) A-line segments. The use of a constant stretch factor for stretching the postcompression A-line is not adequate in the presence of heterogeneous targets that are commonly encountered. This led to the development of several adaptive strain estimation techniques in elastography. Yet, a theoretical framework for the image quality of adaptive strain estimation has not been established. In this work, we develop theoretical expressions for the image quality [measured in terms of the signal-to noise ratio (SNR), contrast-to-noise ratio (CNR) and spatial resolution] of elastograms obtained using an adaptive strain estimator developed by Alam et al. (1998). We show a linear trade-off between the SNR and axial resolution of the strain elastogram with respect to the window length used for strain estimation. The CNR shows a quadratic tradeoff with the axial resolution with respect to the window length. The SNR, CNR and axial resolution are shown to improve with the ultrasonic bandwidth. PMID- 15550323 TI - Image-guided HIFU neurolysis of peripheral nerves to treat spasticity and pain. AB - Spasticity, a major complication of central nervous system disorders, signified by uncontrollable muscle contractions, is very difficult to treat effectively. We report on the use of ultrasound (US) image-guided high-intensity focused US (HIFU) to target and suppress the function of the sciatic nerve complex of rabbits in vivo, as a possible treatment of spasticity. The image-guided HIFU device included a 3.2-MHz spherically curved transducer and an intraoperative imaging probe. A focal acoustic intensity of 1480 to 1850 W/cm(2), applied using a scanning method, was effective in achieving complete conduction block in 100% of 22 nerve complexes with HIFU treatment times of 36 +/- 14 s (mean +/- SD). Gross examination showed blanching of the nerve at the HIFU treatment site and lesion volumes of 2.8 +/- 1.4 cm(3) encompassing the nerve complex. Histologic examination indicated axonal demyelination and necrosis of Schwann cells as probable mechanisms of nerve block. With accurate localization and targeting of peripheral nerves using US imaging, HIFU could become a promising tool for the suppression of spasticity. PMID- 15550324 TI - MRI monitoring of the effect of tissue interfaces in the penetration of high intensity focused ultrasound in kidney in vivo. AB - In this paper, we studied the effect of interfaces during the application of high intensity focused ultrasound (HIFU) ablation in rabbit kidney in vivo. In kidney ablation, mainly two types of interfaces are encountered: these are muscle-kidney and fat-kidney. It was observed that the intensity for which the probability of cavitation (POC) is one was decreased when HIFU penetrated through interfaces, meaning that an interface is a potential site of cavitation. We utilized the concept of scanning the area to be treated in two dimensions (rectangular grid) by applying low intensity ultrasound (diagnostic scan). When all the points of the grid show decrease of signal in T1-weighted fast spoiled gradient (FSPGR) which indicated heating, complete necrosis was observed in the targeted area during the application of HIFU (therapeutic scan). If ultrasound goes through an interface that includes air spaces, the diagnostic scan indicates spaces with poor ultrasound penetration and as a result, during the application of the therapeutic scan, some sites remain untreated. The muscle-kidney and fat-kidney interfaces cause reflection of ultrasound, which prevents the penetration of ultrasound. Microscopic bubbles in the interface may initiate cavitation, especially at high intensities. However, sometimes these types of interfaces do not include any bubbles and therefore the propagation of ultrasound is not inhibited. PMID- 15550325 TI - Activated anti-tumor immunity in cancer patients after high intensity focused ultrasound ablation. AB - T cell-mediated immune responses represent the main cellular antitumor immunity in cancer patients. Recent studies have shown that that both surgical procedure and radiation therapy could cause the functional suppression of lymphocyte mediated cellular immunity. The purpose of current study is to evaluate whether high intensity focused ultrasound (HIFU) might change a systemic antitumor immunity, particularly T lymphocyte-mediated immunity in cancer patients. A total of 16 patients with solid malignancies were treated with HIFU. Among them, six patients had osteosarcoma (Enneking stage, II(B)4, III(B) 2), five had hepatocellular carcinoma (TNM stage, III 3, IV 2), and five had renal cell carcinoma (TNM stage, III 2, IV 3). Using flow cytometry technique, T lymphocyte and subset, B lymphocyte and natural killer cell (NK) in the peripheral blood were measured in these patients on the day before HIFU and 7 to 10 d after HIFU. The statistical significance of any observed difference is evaluated by Student's t-test. The results showed a significance increase in the population of CD4(+) lymphocytes (p < 0.01) and the ratio of CD4(+) /CD8(+) (p < 0.05) in the circulation of cancer patients after HIFU treatment. The abnormal levels of CD3(+) lymphocytes returned toward the normal range in two patients, CD4(+)/CD8(+) ratio in 3, CD19(+) lymphocytes in one and cytotoxic NK in one, respectively, in comparison to control values. It is concluded that HIFU could enhance a systemic antitumor cellular immunity in addition to local tumor destruction in patients with solid malignancies. PMID- 15550326 TI - Biological effects of high-frequency ultrasound exposure during mouse organogenesis. AB - Little has been reported on bioeffects of high-frequency ultrasound (US) and guidelines for US use do not necessarily apply to high frequencies. Pregnant CD-1 mice were exposed to Doppler or B-mode US biomicroscopy (UBM) on embryonic day (E) 8.5 or E10.5, during organogenesis. Operating frequency was 40 MHz with a free field I(SPTA) of 11.9 W/cm(2) (Doppler) and 2.6 mW/cm(2) (B-mode), peak negative pressures of 6.61 MPa and MI of 1.05 (B-mode). Offspring were assessed weekly from 1 day postnatally to euthanasia at 6 weeks, with no significant difference in pup weight, body length or crown-rump length observed. E8.5 Doppler exposed mice showed a small reduction in weight and length at 3 weeks and in weight at 6 weeks. E10.5 Doppler-exposed animals exhibited slight growth reduction in weeks 2 to 4, but were not significantly different at 6 weeks. Our results indicate similar exposures of mice should not cause significant adverse bioeffects. PMID- 15550328 TI - 3-D real-time motion correction in high-intensity focused ultrasound therapy. AB - A method for tracking the 3-D motion of tissues in real-time is combined with a 2 D high-intensity focused ultrasound (US), or HIFU, multichannel system to correct for respiratory motion during HIFU therapy. Motion estimation is based on an accurate ultrasonic speckle-tracking method. A pulse-echo sequence is performed for a subset of the transducers of the phased array. For each of these subapertures, the displacement is estimated by computing the 1-D cross correlation of the backscattered signals acquired at two different times. The 3-D motion vector is then computed by a triangulation algorithm. This technique is experimentally validated in phantoms moving as fast as 40 mm s(-1), and combined with HIFU sequences. A real-time feedback correction of the HIFU beam is achieved by adjusting the delays of each channel. The sonications "locked on target" are interleaved with very short motion-estimation sequences. Finally, in vitro experiments of "locked on target" HIFU therapy are performed in fresh moving tissues. PMID- 15550327 TI - Sonodynamic treatment of murine tumor through second-harmonic superimposition. AB - Acoustic cavitation is the primary mechanism of sonochemical reaction and has potential use for tumor treatment in combination with a certain sonodynamically active agent. It has been known that inducing cavitation with progressive waves is more difficult than with standing waves. This may have been limiting the sonodynamic treatment of tumors. We found that ultrasonically induced chemical reactions are greatly accelerated when the second harmonic is superimposed onto the fundamental. Experimental murine tumors were treated with progressive waves in combination with administration of a gallium-porphyrin complex (ATX-70). The tumors treated with second-harmonic superimposition stopped growing for about 2 days and then gradually started growing again. When only 0.5 MHz was used, tumor growth was not significantly different from that in untreated tumors. It was significantly slower than the untreated when only 1.0 MHz was used, but it was significantly further slowed when second-harmonic superimposition was used. The tumor-bearing mice treated with second-harmonic superimposition after ATX-70 administration survived 5 days longer on average than those untreated. PMID- 15550329 TI - Accreditation for ultrasound in the world. AB - To assess accreditation for ultrasound (US) throughout the world, a questionnaire was sent to 34 doctors in 34 countries; 23 of them responded. Clinical specialties are approved by law in the large majority of countries, except for a few countries like the USA or Japan. The position of "Sonologist", who is a specialist for US, has not been established in any of the countries with the exception of China. In some countries, some kinds of technicians are permitted to be involved in US after training and the recipients of such accreditation are usually called "Sonographers." Accreditation systems for US in various countries were classified into three grades, namely, "Should," "Would" and "Might," according to differences in regulations, education and certification. In 19 out of 23 countries, grading fell into the "Might" grade, in which accreditation is thought to be no more than a matter of morals. It is concluded that a respective accreditation system suitable to the country would be constructed independently. PMID- 15550330 TI - Micromanipulation of endothelial cells: ultrasound-microbubble-cell interaction. AB - Ultrasound (US) in combination with contrast microbubbles has been shown to alter the permeability of cell membranes without affecting cell viability. This permeabilisation feature is used to design new drug delivery systems using US and contrast agents. The underlying mechanisms are still unknown. One hypothesis is that oscillating microbubbles cause cell deformation resulting in enhanced cell membrane permeability. This technical note reveals the interaction between oscillating microbubbles and endothelial cells under a microscope recorded with a fast framing camera at 10 million frames per second. A microbubble expansion of 100% resulted a 2.3-mum displacement of the cell membrane. During the insonification, changes of approximately 15% in the cross-sectional distance of the endothelial cells were observed due to microbubble vibrations. In conclusion, the use of such a camera makes it possible to reveal the mechanisms of interactions between ultrasound, microbubbles and cells. PMID- 15550332 TI - Luteal phase defect: myth or reality. AB - Although the diagnosis of luteal phase defect (LPD) has been described convincingly in the research setting, it remains a controversial clinical entity. Apart from many uncertainties that surround the diagnosis of LPD, there is no convincing evidence that LPD is associated with infertility and recurrent abortion. Once diagnosed, the treatment options are empiric and include those that are recommended for unexplained infertility. The efforts to diagnose LPD in patients who have infertility or recurrent abortion are not justified. PMID- 15550333 TI - Hormonal regulation of implantation. AB - Implantation is a complex process that requires synchronization between the embryo and a receptive endometrium. Hormones, such as the female sex steroids, prostaglandins, and peptide hormones, regulate the cellular and molecular mediators of endometrial receptivity, which include pinopodes, cell adhesion molecules, cytokines, homeobox genes, and growth factors. These mediators can be altered, despite the presence of normal hormone levels and endometrial histology; this limits the usefulness of the luteal phase endometrial biopsy. Therefore, analysis of markers of endometrial receptivity may predict successful implantation better. Elevated androgen and estrogen levels, as seen with polycystic ovary syndrome and controlled ovarian hyperstimulation, respectively, also can have detrimental effects on the endometrium, and therefore, implantation. PMID- 15550334 TI - Hormonal monitoring of the first trimester of pregnancy. AB - The hormonal changes and maternal adaptations of human pregnancy are among the most remarkable phenomena in nature. Endocrinologic parameters in the early gestation period have been used to predict abnormal pregnancies and to identify fetuses that have chromosomal aberrations. This article focuses on the changes in hormones that are secreted by the maternal-fetal-placental unit that are unique for the first trimester of pregnancy and their impact on clinical outcome. PMID- 15550335 TI - Endocrinology of ectopic pregnancy. AB - This article discusses the endocrinology of ectopic pregnancy. PMID- 15550336 TI - Steroid hormone synthesis in pregnancy. AB - The goal of this article is to summarize what is known about the pathways of steroid hormone synthesis and metabolism in human pregnancy. Emphasis is placed on the distinctions between steroidogenic pathways in adults and those that are operative during human pregnancy. PMID- 15550337 TI - Fetal and maternal adrenals in human pregnancy. AB - Human pregnancy is marked by alterations in several endocrine systems--perhaps most notably, the striking increase in steroid hormone production by the adrenals of the fetus and mother. Morphologically and physiologically, the human fetal adrenal glands are remarkable organs. In proportion to the adult organs, the adrenal cortex is the largest organ of the fetus. At term, they produce more steroid and weigh the same as adrenal glands of the adult. Much of the steroid that is released by the fetal and maternal adrenals during pregnancy is the sulfated form of dehydroepiandrosterone (DHEA-S), which is used by the placenta to produce estrogens. Herein, we discuss the physiologic and pathophysiologic hormonal changes of the fetal and maternal adrenals during the course of pregnancy. PMID- 15550338 TI - Fetal hormones and sexual differentiation. AB - The process of fetal sexual differentiation, which involves establishment of genetic sex, differentiation of the gonads, and development of phenotypic sex, is summarized. The morphologic changes that occur in utero that lead to development of the male and female gonads, germ cells, reproductive tracts, and external genitalia are described. Most of the article focuses on the hormones that regulate sexual differentiation and development in utero. The genetic factors that regulate sexual differentiation, which constitute a new and emerging field, also are discussed. PMID- 15550339 TI - Biochemical screening for congenital defects. AB - This article discusses biochemical screening for congenital defects. Biochemical screening remains the standard for estimating the risk of aneuploidy in pregnancy. The combination of first and second trimester markers promises to further improve diagnostic accuracy for anomaly detection. PMID- 15550340 TI - Pituitary gland and pregnancy. AB - The hypothalamic-pituitary-adrenal axis is central to mammalian reproductive function, including conception, pregnancy maintenance, parturition, and breastfeeding. Pregnancy is associated with substantial physiologic changes within this endocrine axis to meet the demands of pregnancy, which include support of the fetus (volume support, nutritional and oxygen supply, clearance of fetal waste), protection of the fetus (from starvation, drugs, toxins), preparation of the uterus for labor, and protection of the mother from potential cardiovascular injury at delivery. This article reviews the anatomy, embryology, and physiology of the pituitary. The effect of pregnancy on pituitary structure and function, in health and disease, also is discussed. PMID- 15550341 TI - Thyroid disease in pregnancy. AB - Maternal, fetal, and neonatal thyroid physiology are discussed. Moreover, this article serves as a review of the more common thyroid diseases that are encountered during pregnancy and the postnatal period, their treatments, and their potential effects on pregnancy. PMID- 15550342 TI - Diabetes mellitus in pregnancy. AB - This article reviews normal and abnormal carbohydrate metabolism in pregnancy, with an emphasis on the challenges that are faced by those who care for the pregnant woman who has hyperglycemia. The growing problem of type 2 diabetes in pregnancy, the controversial use of oral antihyperglycemic agents for the treatment of gestational diabetes, and the long-term issue of diabetes prevention in those whose hyperglycemia resolves postpartum are also addressed. PMID- 15550343 TI - The initiation of parturition at term. AB - This article discusses the initiation of parturition at term. PMID- 15550344 TI - Effects of hormones on fetal lung development. AB - Endogenous hormones, such as glucocorticoids, play a major role in the development of the fetal lung. Considerable effort has been devoted to defining the underlying physiology and the clinical effects of administration of antenatal glucocorticoids to women who are at risk of premature delivery. Antenatal glucocorticoids have significant therapeutic benefits to the neonate with respect to the respiratory distress syndrome, intraventricular hemorrhage, and mortality. Current controversies relate to the choice of glucocorticoid, optimal dosing regime, number of courses of therapy that should be administered, and, most importantly, potential toxicities. PMID- 15550345 TI - Endocrinology of lactation. AB - The endocrine control of lactation is one of the most complex physiologic mechanisms of human parturition. Mammogenesis, lactogenesis, galactopoiesis, and galactokinesis are all essential to assure proper lactation. Prolactin is the key hormone of lactation and seems to be the single most important galactopoietic hormone. Oxytocin, serotonin, opioids, histamine, substance P, and arginine leucine modulate prolactin release by means of an autocrine/paracrine mechanism, whereas estrogen and progesterone hormones can act at the hypothalamic and adenohypophysial levels. Human placental lactogen and growth factors play an essential role to assure successful lactation during pregnancy. Oxytocin is the most powerful galactokinetic hormone. PMID- 15550346 TI - Alterations of neuroplasticity in depression: the hippocampus and beyond. AB - Early hypotheses on the pathophysiology of major depression were based on aberrant intrasynaptic concentrations of mainly the neurotransmitters serotonin and norepinephrine. However, recent neuroimaging studies have demonstrated selective structural changes across various limbic and nonlimbic circuits in the brains of depressed patients. In addition, postmortem morphometric studies revealed decreased glial and neuron densities in selected brain structures supporting the idea that major depression may be related to impairments of structural plasticity. Stressful life events are among the major predisposing risk factors for developing depression. Using the chronic psychosocial stress paradigm in male tree shrews, an animal model with a high validity for the pathophysiology of depressive disorders, we found that 1 month of stress reduced the in vivo concentrations of the brain metabolites N-acetyl-aspartate, choline containing compounds, and (phospho)-creatine, as well as the proliferation rate in the dentate gyrus and the hippocampal volume. Even though long-lasting social conflict does not lead to a loss of principal cells, the hippocampal changes were accompanied by modifications in the incidence of apoptosis. Notably, these suppressive effects of social conflict on hippocampal structure could be counteracted by treatment with the antidepressant tianeptine. These findings support current theories proposing that major depressive disorders may be associated with impairment of structural plasticity and neural cellular resilience, and that antidepressants may act by correcting this dysfunction. PMID- 15550347 TI - Preclinical research on stress, memory, and the brain in the development of pharmacotherapy for depression. AB - We have reviewed two areas of research on stress, memory, and synaptic plasticity which may be relevant toward understanding the neurobiology of major depressive disorder (MDD). First, we have presented the view that the hippocampus (HC) and prefrontal cortex (PFC) function jointly as a memory system which enables multitask processing (working memory) and consolidation of contextual information. The amygdala, by contrast, is necessary for the consolidation of emotional memories. Cognitive and neurophysiological studies have shown that HC PFC processing is impaired, and amygdaloid processing is enhanced, by stress and in anxiety and mood disorders, including MDD. Second, we have reviewed research on the behavioral and neurophysiological actions of tianeptine, an antidepressant that is known to block the adverse effects of chronic stress on hippocampal morphology. Recent work has shown that acute tianeptine enhances cognitive and electrophysiological measures of HC-PFC functioning without interfering with the emotion-induced enhancement of amygdaloid functioning in rodents. We conclude with a synthesis of the preclinical and clinical literature on stress, memory, and tianeptine with the proposal that tianeptine should enhance HC-PFC memory related processing in people under stress. PMID- 15550348 TI - Molecular mechanisms of neuroplasticity and pharmacological implications: the example of tianeptine. AB - The hippocampal formation, which expresses high levels of adrenal steroid receptors, is a malleable brain structure that is important for certain types of learning and memory. This structure is also vulnerable to the effects of stress hormones which have been reported to be increased in depressed patients, particularly those with severe depression. The amygdala, a structure that plays a critical role in fear learning, is also an important target of anxiety and stress. Certain animal models of depression involve application of repeated stress. Repeated stress promotes behavioral changes that can be associated with these two brain structures such as impairment of hippocampus-dependent memory and enhancement of fear and aggression, which are likely to reflect amygdala function. At a cellular level, opposite responses in the hippocampus and amygdala are observed, namely, shrinkage of dendrites in hippocampus and growth of dendrites in the lateral amygdala, involving in both cases a remodeling of dendrites. Furthermore, stress-induced suppression of neurogenesis has been noted in dentate gyrus. At a molecular level, the effects of repeated stress in the hippocampus involve excitatory amino acids and the induction of the glial form of the glutamate transporter. Chronic treatment with the antidepressant tianeptine may prevent these effects in hippocampus and amygdala. PMID- 15550349 TI - Neuroplasticity: from MRI to depressive symptoms. AB - Morphological and functional changes have been repeatedly reported in the brain organization of depressed patients. The main modifications demonstrated by structural magnetic resonance imaging (MRI) are a reduction in the gray matter volume within the prefrontal cortex, the hippocampus, and the striatum. The reduction in gray matter volume and the morphological atrophy are probably due to an excess of neural loss (apoptosis) and an altered regulation of the neurotrophic processes. Hence, a deficit in neurotrophic factor synthesis (brain derived neurotrophic factor [BDNF], neurotrophin [NT]-3, NT-4/5, Bcl-2, etc.) may be responsible for increased apoptosis in the hippocampus and prefrontal cortex corresponding to the cognitive impairment described in depression. This hypothesis seems to be confirmed by the decreased expression of neurotrophic factors (e.g., BDNF mRNA) in animal models of depression. In parallel, the neural plasticity (functional aspects of synaptic connectivity and long-term potential activity [LTP]) is decreased. However, the most interesting data concern the possible reversibility of this dysregulation with antidepressant treatment. For example, communication between the hippocampus and the prefrontal cortex could be re-established, enabling in a way the cognitive processes to be "reset." From a clinical point of view, the consequences of such a phenomenon are manifold: PMID- 15550350 TI - From restoration of neuroplasticity to the treatment of depression: clinical experience. AB - The adult brain has more plasticity than previously believed. Neurogenesis, growth and branching of dendrites, and remodeling of synaptic contacts in different regions of the brain occur continuously. Numerous studies have reported a decrease in neuroplasticity in depressed patients and/or in animals subjected to stress and to different models of depression. This has led to the proposal of a new approach to the pathophysiology of depression: depression could be the result of the decrease in neuroplasticity in brain structures involved in the control of mood. This new approach to the pathophysiology of depression can lead to better understanding of, or the proposal of more solid hypotheses about, some issues such as the impact of genetics and environmental factors on the occurrence of depressive episodes, the increased risk of depression in patients with somatic diseases in which there are alterations of neuroplasticity, or the increased risk of depressive relapse in depressed patients in partial remission in whom we suspect that neuroplasticity is only partially restored. These observations have also led to the proposal of new hypotheses concerning the mode of action of antidepressant drugs. In this regard, tianeptine is of particular interest. Tianeptine's pharmacological and clinical properties have been extensively studied. Tianeptine has specific neurotrophic properties, and its antidepressant properties have been well demonstrated. Tianeptine provides early relief of anxious symptoms without sedation in depressed patients. The acceptability and safety profiles of tianeptine are appreciated by both physicians and patients; for instance, tianeptine does not induce sexual dysfunction, nausea, or weight gain. It is of interest to focus on what we already know about tianeptine's pharmacological and clinical properties, and to create mechanistic hypotheses about the similarities and differences observed in clinical practice between tianeptine and other antidepressants. PMID- 15550352 TI - Transethnic itineraries for ethnomedical therapies in Nigeria: Igbo women seeking Ibibio cures. AB - Although therapeutic itineraries have been studied in a variety of contexts, little research has investigated care-seekers' quests for traditional medical treatments outside their own ethnic boundaries. The present study investigated 19 Igbo women seeking traditional cures from Ibibio indigenous healers in Akwa Ibom State, Nigeria. Emerging data show that these Igbo women were uptaking Ibibio indigenous treatments primarily for health conditions that have failed to respond to initial treatments in their places of origin, were stigmatized at their own places of origin, or and were thought to have resulted from supernatural causes. Care-seeking outside patients' ethnic borders was thus both a quest for a more effective treatment and a strategy for concealing therapeutic progress or and the nature of illness from the patients' places of origin. Findings underscore the critical role of culture and place in health-seeking behaviour and the need for health care services to be responsive to the complex nature of cultural organization involving care-seekers' and the critical ways this plays out in, flows into, and is negotiated through particular places during therapeutic quests. PMID- 15550353 TI - The importance of place of residence in predicting late-stage diagnosis of breast or cervical cancer. AB - We tested whether inner-city women were at significantly increased risk of late stage cancer diagnosis because they resided in extremely poor and socially isolated neighborhoods or in neighborhoods meeting the federal definition of a medically underserved area (MUA). Cancer registry data on women in three American cities were matched to Census data. Using logistic regression we found that residence in economically and socially distressed or medically underserved neighborhoods tended to increase the likelihood of late-stage cancer diagnoses. Further, we found that not all areas that are economically and socially distressed receive the federal MUA designation. Consequently, we argue that economically and socially distressed neighborhoods should be automatically designated as MUA. PMID- 15550354 TI - The farm as a setting for health promotion: results of a needs assessment in South Central Alberta. AB - Farming is acknowledged to be a dangerous occupation that poses risks for farmers, farm employees and family members. This paper explores the farm as a setting where health promotion and prevention programs can be implemented. We present data from an occupational health and safety needs assessment of 347 farms in the South Central region of the province of Alberta, Canada. The data are used to describe farmers' familiarity with farm living, thoughts on farm safety, the numbers of people working and types of work performed, other work, income, and the availability of resources. Results are discussed in terms of implications for practitioners under three proposed justifications for a settings model. Data revealed that there is so much heterogeneity among these farms that an ecological settings approach to program development that accounts for both the composition and context of rural settings is more appropriate than the settings model currently in use. PMID- 15550355 TI - Increasing inequalities in risk of murder in Britain: trends in the demographic and spatial distribution of murder, 1981-2000. AB - This study analyses demographic and spatial factors that underlie the rise in murder rates seen in Britain between 1981 and 2000 and considers the possible contribution of a public health approach to the understanding of murder. Comparison of murder rates by age group and sex finds that increases occurred only among males aged 5-59 years, and were greatest among males aged 20-24 years. Analysis of the relationship with poverty at the area level, using the Breadline Britain index and deciles based on wards, demonstrates that increases in murder rates were concentrated in the poorest areas. Rates of murder have risen in the same population groups and areas that have experienced increases in suicide and may be associated with worsening social and spatial inequality. PMID- 15550356 TI - Is inter-school variation in smoking uptake and cessation due to differences in pupil composition? A cohort study. AB - The aims of this study were to determine if inter-school variation in smoking prevalence was due to differences in pupil composition or other school-level factors. A cohort of 13-14-year-olds (n = 7147) from 52 schools was followed-up 1 year later. Random effects logistic regression was used to examine school variation in smoking uptake and cessation, with and without adjustment for pupil composition. Inter-school variation in smoking prevalence is not caused by differences in pupil composition but is due to differences in the onset of smoking arising because of unmeasured school contextual or collective factors operating on pupils' decisions. PMID- 15550357 TI - Lessons learned from malaria: Italy's past and sub-Sahara's future. AB - No longer a major public health concern in developed countries, malaria kills 1-3 million people annually, mostly children under the age of five in sub-Saharan Africa. In 1998, the WHO launched the Roll Back Malaria (RBM) drive to halve malaria mortality by 2010. This article contrasts the problems confronting RBM with the successful Italian drive to eradicate malaria between the late 19th and mid 20th centuries. The Italians employed education and applied socio-political will; however, ecological and socio-economic conditions in sub-Saharan Africa are more hospitable to the disease. RBM strategies should consider the Italian experience while awaiting a major scientific breakthrough necessary to achieve success. PMID- 15550358 TI - Supportive and palliative care for patients from ethnic minorities in Europe: do we suffer from institutional racism? PMID- 15550359 TI - Creating a culture of compassion: developing supportive care for people with cancer. AB - Receiving the 2003 Distinguished Merit Award from the European Oncology Nursing Society is a great moment in my professional career. It is also a time for reflection. We can easily become immersed in the specifics of our work and forget the larger picture. An opportunity such as this allows me to step back, reflect and review what I have accomplished. It also challenges me to think about the totality of cancer nursing and cancer care, look at where we have been and about where we should be going. At the heart of this lies the concept of supportive care. I would like to consider three pertinent areas. First, I will define what I consider the domain of supportive care in cancer to be. Second, I will look at what is needed to further supportive care for people with cancer. This involves building the discipline of more rigorous symptom assessment and documentation; better management of the symptoms and concerns that confront people with cancer; moving beyond the traditional framework of treatment and care, embracing a more integrated approach; addressing quality whilst at the same expediting the delivery of supportive care services. Third, I would like to consider the challenges to reform that this presents for cancer nursing and cancer nurses. A road map for change will be presented which highlights both the necessity to promote a supportive care culture whilst simultaneously building a dedicated infrastructure of staff and services. Nurses must play a key role in supportive care. Because of our unique clinical and research base, we are primed to assume leadership roles in both these spheres. Mutual valuing, partnership and shared working are the only means of delivering enhanced cancer care. We should grasp opportunities, confident that together we have the skills and knowledge to move forward. Today is yesterday's tomorrow. We cannot do anything about yesterday, but we can do something about today to ensure tomorrow is how we want it to be. We can become what we dream, let us live that dream outside and really drive forward the care we provide for people with cancer and their families. PMID- 15550360 TI - The professional role of breast cancer nurses in multi-disciplinary breast cancer care teams. AB - Since the 1970s breast cancer services have witnessed considerable changes in the management of patients. One significant change was the introduction of specialist core personnel, including the breast care nurse (BCN). The role of the BCN has been gaining credence rapidly in the British NHS and this service is perhaps the paradigm of care for other services. With the lack of specific evidence of the role of specialist nurses in the breast care team, the current study aims to explore this area by in-depth interviews with core team members, and observations of 16 multi-disciplinary teams in England. The study explores the following themes: Nurses' unique informal management leadership role in ensuring the co ordination, communication and planning of the team work; nurses' innovatory role in making the bureaucracy respond to patients and their relatives needs; nurses supportive role in the provision of expert advice and guidance to other members of the team; nurses confidence and humour in well-performing teams; and the limitations of the professional role of the breast cancer nurse. This study indicates that there is evidence that the BCN is practicing at an advanced level of practice. However, there is a severe lack of evidence-based description of that advanced practice. Cancer nurses including the BCNs should develop and participate in programmes of research in line with cancer legislation in order to build an evidence base that ultimately supports their unique role. PMID- 15550361 TI - The information needs of women who have undergone breast reconstruction. Part II: Information giving and content of information. AB - Women diagnosed with breast cancer treated by mastectomy can choose breast reconstruction. The information needs of women undergoing this procedure have only been addressed in the research literature to a limited extent. A qualitative approach was used to explore the experiences of women who had undergone breast reconstruction with a specific focus on their views on how they considered their information needs could best be met. A purposeful sample of eight women was recruited to participate in two focus groups, each lasting 2 hours. Framework analysis was used to develop an index of key themes and sub themes which transformed the data into a structured record which facilitated systematic analysis. This paper will present the emergent key themes regarding information giving and the content of information that women perceive as important when preparing for breast reconstruction. Process, delivery and patient factors are presented in the category of information giving. Several sub themes are discussed concerning the content of information considered to be relevant. Those involved in imparting information to women about such surgery should be aware of the type of information that is considered relevant, the manner in which it should be delivered and timing factors that implicate on the process. PMID- 15550362 TI - Telephone first post-intervention follow-up for men who have had radical radiotherapy to the prostate: evaluation of a novel service delivery approach. AB - The increasing prevalence of prostate cancer places pressure on services, leading to questions about how best to configure services, so as to maintain quality and best utilise the skills of the multi-disciplinary team. There have been positive evaluations of specialist nursing roles, nurse-led service provision and telephone consultations, leading us to consider whether telephone follow-up led by a specialist nurse might be an acceptable alternative to traditional follow-up in hospital-based out-patient clinics for patients receiving radical radiotherapy for prostate cancer. Thirty-six men were included in a pilot introduction of telephone follow-up, evaluated via a questionnaire survey. The evaluation explored patients' satisfaction with practical arrangements; satisfaction with the nurse; acceptability of telephone follow-up and acceptability of this being nurse-led. Patients reported high levels of satisfaction with practical arrangements. Thirty-five patients considered the nurse to be knowledgeable and found nurse-led care acceptable. Only one patient was unhappy with telephone follow-up, whilst 27 were "very happy". Only one patient felt that telephone follow-up was poorer than traditional follow-up, whilst 27 found it "as good" and three "better". Particular advantages were reported in terms of convenience and time savings. Telephone follow-up appears to have potential for this patient group and merits wider, research-based consideration. PMID- 15550363 TI - Fatigue patterns in Chinese patients receiving radiotherapy. AB - Fatigue has been reported as the most frequently occurring symptom in cancer patients receiving chemoradiotherapy. The purpose of the current descriptive study was to explore the pattern, associated factors, and experience of fatigue in Chinese cancer patients receiving radiotherapy. Twenty-seven subjects from an out patient clinic of a university hospital in Hong Kong participated in the study. They were asked to complete three times daily (morning, afternoon and evening) a diary over a period of two weeks from the beginning of their therapy. The diary incorporated a 10-cm horizontal visual analogue scale (VAS) for measuring fatigue intensity. Further, the Chinese version of the fatigue subscale of the Profile of Mood States was completed before the radiotherapy, at the end of the first week post-treatment, and at the end of the second week. Most patients (44-80.8%) undergoing radiotherapy in this study experienced fatigue at some point during their treatment. The pattern of fatigue was that of repeated curves, with higher levels observed in the afternoons and evenings, and at the end of the first week or the end of the second week. Qualitative data indicated the universal sensations of fatigue and also some culturally specific descriptors. A variety of methods were used by the patients to cope with their fatigue but the effects of such methods were not always positive. It is important for clinical practice to assess the individual patterns and perceptions of fatigue, identify those patients at risk for developing fatigue and initiate interventions to combat fatigue. PMID- 15550364 TI - Children's acceptance and tolerance of chlorhexidine and benzydamine oral rinses in the treatment of chemotherapy-induced oropharyngeal mucositis. AB - Oral care is of great importance in the prevention of chemotherapy-induced oropharyngeal mucositis. Although considerable attention has been given in improving oral care practices, patients' acceptance and tolerance of oral rinses is a continuing problem in oral care. A randomized crossover design was used to determine the relative acceptability and tolerability of chlorhexidine and benzydamine oral rinse agents in children receiving chemotherapy. At the end of the study, each subject was asked to compare these two agents in relation to stinging and taste, as well as his/her perception in reducing mucositis. Thirty four children aged 6-17 years completed two courses of chemotherapy during which they alternately practiced oral care using chlorhexidine then benzydamine or benzydamine then chlorhexidine. All of the children tolerated the agents well and continued with rinsing throughout the study. Only a few children had to resort to diluting the agents with normal saline or water. Fifty-nine percent of children reported that the stinging associated with benzydamine was more accepted than chlorhexidine. The taste of both these agents was accepted by 50% of children. Approximately 60% of children reported that chlorhexidine was more helpful than benzydamine in reducing mucositis. About 47% and 50% of them preferred chlorhexidine and benzydamine in their subsequent chemotherapy, respectively. In conclusion, chlorhexidine and benzydamine are acceptable and well-tolerated by children over the age 6 years old. PMID- 15550365 TI - Improving patient access to healthcare professionals: a prospective audit evaluating the role of e-mail communication for patients with lung cancer. PMID- 15550366 TI - Extravasation. PMID- 15550367 TI - Involvement of chloride in apoptotic cell death induced by activation of ATP sensitive P2X7 purinoceptor. AB - The ATP-gated P2X(7) receptor is a plasma membrane receptor belonging to the family of P2X purinoceptors. Its activation leads to multiple downstream events including influx of ions, pore formation to allow the passage of larger molecular weight species, and cell death by apoptosis and/or necrosis. The cell death is thought to be correlated with the pore formation but does not directly result from the dilatation of pores. We have generated and characterized a clone of chicken DT40 lymphocytes stably transfected with the rat P2X(7) receptor. In this study, we investigated the mechanism of P2X(7) receptor-induced cell death using this clone. Treatment with P2X(7) receptor agonist, 2'-3'-O-(4-benzoylbenzoyl) ATP induced depolarization of membrane potential, pore formation, and cell shrinkage, an early hallmark of apoptosis in the buffer containing physiological concentrations of ions. Analysis by flow cytometry revealed that the activity of pore formation in shrunk cells was much higher than in non-shrunk cells. The activation of P2X(7) receptor also caused the release of lactate dehydrogenase from cells. The P2X(7) receptor-mediated cell shrinkage and lactate dehydrogenase release were blocked when media Cl(-) was replaced with gluconate. However, removal of extracellular Cl(-) did not affect plasma membrane depolarization and pore formation by treatment with 2'-3'-O-(4-benzoylbenzoyl)-ATP. Therefore we concluded that pore formation plays a critical role in the P2X(7) receptor induced apoptotic cell death and that this is mediated by extracellular Cl(-) influx. PMID- 15550368 TI - Five year follow up of biocolonisable microporous fluorocarbon haptic (BIOKOP) keratoprosthesis implantation in patients with high risk of corneal graft failure. AB - AIM: To study the anatomical and visual performance following implantation of a model of artificial cornea and to evaluate the postoperative long term complications. METHODS: 11 eyes of 11 patients with bilateral corneal blindness considered as potentially having high risk of failure of penetrating corneal keratoplasty were implanted with biocolonisable Kpro keratoprosthesis (BIOKOP I, FCI, Rantigny, France) in the period between January 1996 and May 1998. Only one eye was implanted in all patients and followed up for a period of 60 months. The visual outcome, anatomical and functional stability, complications, and the general performance of the keratoprosthesis were evaluated. RESULTS: The keratoprosthesis (BIOKOP I) only 36.3% remained in position to date. In the patients' last visit five eyes (45.4%) were blind and one (9.0%) showed a slight improvement in the best corrected visual acuity (BCVA) in comparison to preoperative tests. Six eyes (54.5%) showed improved BCVA before having postoperative complications. Four eyes underwent replacement of a BIOKOP I Kpro with a BIOKOP II as a result of extrusion. The keratoprostheses remained anatomically in situ for a mean of 25.5 months and their functional performance period was limited to a mean of 22 months. CONCLUSION: Corneal keratoprosthesis (BIOKOP I, II) does not provide a stable anatomical relation with the surrounding ocular structures. Its ability to restore vision is limited to a short postoperative period in eyes implanted with severe ocular surface disease. PMID- 15550369 TI - Cryotherapy for breast fibroadenomas. AB - PURPOSE: To assess freezing protocols, imaging, and clinical outcomes of percutaneous ultrasonographically (US)-guided cryotherapy for breast fibroadenomas. MATERIALS AND METHODS: Institutional review board approval and patient consent were obtained. Forty-two biopsy-confirmed fibroadenomas were treated in 29 patients (mean age, 27 years) by using a 2.4-mm cryoprobe inserted into the fibroadenoma with US guidance. The first seven patients underwent conscious sedation, but the other 22 patients required only local anesthesia. US and thermocouple monitoring of the procedure were performed to evaluate freeze protocols based on tumor size. Saline injections protected the skin and/or chest wall. US follow-up was performed at 1 week and at 1, 3, 6, and 12 months. Pre- and 12-month postcryotherapy mammograms were available for seven patients who were over 30 years old. chi(2) and Student t tests were used to assess frequency and mean differences, respectively. RESULTS: The 22 patients who underwent local anesthesia reported minimal discomfort. No significant complications were noted, and patients were very pleased with the resolution of palpable mass effect and cosmetic results. The average pretreatment fibroadenoma volume of 4.2 cm(3)+/- 4.7 (standard deviation) was reduced to 0.7 cm(3)+/- 0.8 at 12-month follow-up (73% reduction, P < .001). US produced excellent ice visualization beyond tumor margins, while thermocouples confirmed cytotoxic temperatures approximately 5 mm behind the visible leading edge. Two patients elected to undergo either removal or biopsy of a residual mass, which revealed a shrunken hyaline matrix with preserved collagenous architecture. Mammograms showed comparable resolution of mass effects with mild surrounding parenchymal reaction. CONCLUSION: Cryotherapy of fibroadenomas is a safe, effective, and virtually painless clinic-based (ie, outpatient) treatment option with good cosmesis. PMID- 15550370 TI - Acute lung injury: effects of prone positioning on cephalocaudal distribution of lung inflation--CT assessment in dogs. AB - PURPOSE: To quantify cephalocaudal gradient of lung inflation in acute lung injury in a dog model in prone versus supine position. MATERIALS AND METHODS: Experiments were performed in accordance with Guide for the Care and Use of Laboratory Animals, as approved by National Research Council (National Institutes of Health), and were approved by committee on care and use of animals in research at Seoul National University Hospital. After induction of acute lung injury with intravenous injection of oleic acid, dogs were randomized to be ventilated in either prone (n = 6) or supine (n = 6) position. Spiral computed tomography (CT) and hemodynamic measurement were performed sequentially on an hourly basis. Volume and mean attenuation of lung were measured quantitatively by using software to evaluate each CT section. Cephalocaudal gradient of mean lung attenuation, distribution of gas and tissue, and alveolar expansion were assessed. Functional residual capacity and net alveolar expansion of entire lung were measured. Statistical analysis was performed with Friedman, sign, and Mann Whitney tests. RESULTS: Mean lung attenuation increased gradually from apex to base of lung in supine position. Thus, inflation gradient along cephalocaudal axis was found. Gas was located predominantly in upper lung, whereas tissue was dominant in lower lung in supine position. In supine group, cephalocaudal inflation gradient showed no significant change from baseline up to 4 hours. After prone positioning, cephalocaudal inflation gradient was reduced, and gas and tissue proportions became more uniform along cephalocaudal axis. In prone group, absolute values of cephalocaudal inflation gradient at time points of prone positioning for 1, 2, and 3 hours were significantly lower than baseline values (P < .05) and those in supine group (P < .05). Alveolar expansion occurred in caudal regions, and alveolar contraction occurred in cephalic regions; accordingly, net alveolar volume of entire lung was not altered significantly. Functional residual capacity was unchanged by prone positioning. CONCLUSION: In acute lung injury, prone positioning induced more uniform distribution of gas and tissue along cephalocaudal axis by reducing cephalocaudal inflation gradient. PMID- 15550371 TI - Coronary artery stenosis: direct comparison of four-section multi-detector row CT and 3D navigator MR imaging for detection--initial results. AB - PURPOSE: To prospectively compare the diagnostic accuracy of multi-detector row computed tomography (CT) and of three-dimensional (3D) navigator magnetic resonance (MR) imaging in patients referred for conventional coronary angiography for detection of coronary artery stenosis. MATERIALS AND METHODS: All patients gave written informed consent for the study, which was approved by the local ethics committee. Twenty-seven patients underwent multi-detector row CT and 3D navigator free-breathing MR imaging a mean of 5 days before undergoing invasive coronary angiography. The acquired multi-detector row CT and MR images were graded for the presence of greater than 50% stenosis in vessels larger than 1.5 mm in diameter. The diagnostic accuracies of the two examinations were compared with that of quantitative coronary angiography (QCA) by using the McNemar test. RESULTS: Owing to claustrophobia, MR images were not acquired in one patient; thus, 26 patients were included for analysis. According to QCA findings, 21 of the 26 patients had significant coronary artery disease and 58 (20%) of a total of 294 coronary artery segments larger than 1.5 mm in diameter had significant (>50%) stenosis. Multi-detector row CT had significantly higher sensitivity (46 [79%] of 58 segments) than MR imaging (36 [62%] segments, P < .05) for detection of segments with significant stenosis. Conversely, MR imaging had significantly higher specificity (198 [84%] of 236 segments) than did CT (168 [71%] segments, P < .001) for exclusion of segmental coronary artery stenosis. Both examinations had high negative predictive value for exclusion of segmental stenosis: 93% (168 of 180 segments) for CT and 90% (198 of 220 segments) for MR imaging. The overall diagnostic accuracy of MR imaging (80% [234 of 294 segments]) was significantly higher than that of CT (73% [214 segments], P < .05). CONCLUSION: MR imaging had significantly higher diagnostic accuracy than multi-detector row CT in the evaluation of coronary artery stenosis. Both techniques have high negative predictive value, making them particularly useful for ruling out coronary artery disease in symptomatic patients. PMID- 15550372 TI - Rectal cancer: MR imaging in local staging--is gadolinium-based contrast material helpful? AB - PURPOSE: To determine retrospectively whether addition of gadolinium-enhanced T1 weighted magnetic resonance (MR) sequence to T2-weighted turbo spin-echo (SE) MR imaging is valuable for preoperative assessment of T stage and circumferential resection margin in patients with primary rectal cancer. MATERIALS AND METHODS: Local institutional review board approved study and waived informed patient consent. Eighty-three patients with operable primary rectal cancer underwent preoperative MR imaging. Retrospectively, two observers independently scored T2 weighted turbo SE MR images and, in a second reading, T2-weighted images combined with gadolinium-enhanced T1-weighted turbo SE MR images for tumor penetration through rectal wall and tumor extension into mesorectal fascia. A confidence level scoring system was used, and receiver operating characteristic (ROC) curves were generated. Histologic findings were standard of reference. Difference in performance of T2-weighted and combined T2-weighted plus gadolinium-enhanced T1 weighted sequences was analyzed by comparing corresponding areas under ROC curves (A(z)) for each observer. Interobserver agreement was calculated by using linear weighted kappa statistics. RESULTS: Addition of contrast-enhanced T1-weighted to T2-weighted MR imaging did not significantly improve diagnostic accuracy for prediction of tumor penetration through rectal wall (A(z) of T2-weighted vs T2 weighted plus T1-weighted images for observer 1, 0.740 vs 0.764; observer 2, 0.856 vs 0.768) and tumor extension into mesorectal fascia (A(z) for observer 1, 0.962 vs 0.902; observer 2, 0.902 vs 0.911). Diagnostic performance (A(z)) of MR and interobserver agreement were high for prediction of tumor extension into mesorectal fascia (kappa = 0.61, 0.74) but only moderate for penetration through rectal wall (kappa = 0.47, 0.45). CONCLUSION: Gadolinium-enhanced MR sequences did not improve diagnostic accuracy for assessment of tumor penetration through rectal wall and tumor extension into mesorectal fascia. PMID- 15550373 TI - Noninvasive coronary angiography with 16-detector row CT: effect of heart rate. AB - PURPOSE: To evaluate the effect of heart rate on the quality of coronary angiograms obtained with 16-detector row computed tomography (CT) by using temporally enhanced three-dimensional (3D) approaches. MATERIALS AND METHODS: The local ethics committee approved the study, and informed consent was obtained from all patients. Fifty patients underwent coronary CT angiography (heart rate range, 45-103 beats per minute). Raw data from helical CT and electrocardiography (ECG) were saved in a combined data set. Retrospectively ECG-gated images were reconstructed at preselected phases (50% and 80%) of the cardiac cycle. A 3D voxel-based approach with cardiac phase weighting was used for reconstruction. Testing for correlation between heart rate, cardiac phase reconstruction window, and image quality was performed with Kruskal-Wallis analysis. Image quality (freedom from cardiac motion-related artifacts) was referenced against findings at conventional angiography in a secondary evaluation step. Regression analysis was performed to calculate heart rate thresholds for future beta-blocker application. RESULTS: A significant negative correlation was observed between heart rate and image quality (r = 0.80, P < .001). Motion artifact-free images were available for 44 (88%) patients and were achieved consistently at a heart rate of 80 or fewer beats per minute (n = 39). Best image quality was achieved at 75 or fewer beats per minute. Segmental analysis revealed that 97% of arterial segments (diameter > or = 1.5 mm according to conventional angiography) were assessable at 80 or fewer beats per minute. Premature ventricular contractions and rate-contained arrhythmia did not impede diagnostic assessment of the coronary arteries in 10 (83%) of the 12 patients affected. CONCLUSION: Motion free coronary angiograms can be obtained consistently with 16-detector row CT scanners and adaptive multicyclic reconstruction algorithms in patients with heart rates of less than 80 beats per minute. PMID- 15550374 TI - MR imaging of the wrist: comparison between 1.5- and 3-T MR imaging--preliminary experience. AB - Institutional review board approval and informed consent were obtained from 25 healthy volunteers and 15 consecutive patients with chronic wrist pain or suspected carpal mass, and 1.5- and 3-T magnetic resonance (MR) imaging of the wrist was prospectively performed with comparable sequence parameters and surface coils of the same geometric design. Imaging protocols at both field strengths included a T1-weighted spin-echo sequence, two intermediate-weighted fast SE sequences with different echo times and with and without fat saturation, and a three-dimensional fast field-echo sequence. The contrast-to-noise ratio (CNR) between muscle and bone and between bone and cartilage was calculated for both field strengths. The visibility of various anatomic structures, including the triangular fibrocartilage complex, carpal ligaments, nerves, and cartilage, was analyzed with a four-point grading scale. CNRs were significantly higher on 3-T MR images than on 1.5-T MR images (P < .001; analysis of variance) for all sequences. Visibility of the triangular fibrocartilage complex and intercarpal ligaments and cartilage was significantly better on 3-T MR images than on 1.5-T MR images (paired sign test). PMID- 15550375 TI - Hemoptysis: bronchial and nonbronchial systemic arteries at 16-detector row CT. AB - PURPOSE: To retrospectively evaluate 16-detector row computed tomography (CT) compared with conventional angiography in depiction of bronchial and nonbronchial systemic arteries in patients with hemoptysis. MATERIALS AND METHODS: Institutional review board approval was obtained, and informed consent was not required. Sixteen-detector row helical CT and conventional angiography of the thorax were performed in 22 patients (16 men, six women; age range, 18-75 years; mean age, 50 years) with hemoptysis. Three observers in consensus analyzed retrospectively transverse, multiplanar reconstruction, or three-dimensional CT images for visibility, traceability of bronchial arteries from their origin at the aorta or aortic branches to the hilum, and presence of nonbronchial systemic arteries. CT and angiographic findings of bronchial and nonbronchial systemic arteries causing hemoptysis were compared by two radiologists in consensus. Differences in visibility, traceability, and diameter of bronchial arteries causing and those not causing hemoptysis were tested by using generalized estimating equation method or the mixed model. RESULTS: Fifty-two (30 right and 22 left) bronchial arteries and 33 nonbronchial systemic arteries were visible at CT. Thirty-four (20 right and 14 left) of 52 bronchial arteries were traceable from their origins to the hilum. Thirty-one (16 right and 15 left) of 46 (27 right and 19 left) bronchial arteries and 26 of 64 nonbronchial systemic arteries evaluated at angiography were causing hemoptysis. Forty (87%, 23 right and 17 left) of 46 bronchial arteries seen at angiography were also detected at CT. All 31 bronchial arteries and sixteen (62%) of 26 nonbronchial systemic arteries causing hemoptysis were detected at CT. Twenty-three (74%) of 31 bronchial arteries causing hemoptysis were traceable from their origins to the hilum, and one (11%) of nine bronchial arteries not causing hemoptysis was traceable (P = .002). CONCLUSION: Sixteen-detector row CT provides depiction and traceability of the bronchial arteries in patients with hemoptysis, and in most patients it enables detection of the bronchial and nonbronchial arteries causing hemoptysis. PMID- 15550376 TI - Change in knee cartilage T2 at MR imaging after running: a feasibility study. AB - All participants provided informed consent to participate in this study, which was approved by the institutional review board of Milton S. Hershey Medical Center. The purpose of the study was to determine the feasibility of cartilage T2 mapping in the evaluation of response of femoral and tibial cartilage to running exercise. Quantitative magnetic resonance (MR) T2 maps of weight-bearing femoral and tibial articular cartilage were obtained in seven young healthy men before and immediately after 30 minutes of running by using a 3.0-T MR imager. There was no statistically significant change in T2 profiles of tibial cartilage. There was a statistically significant decrease in T2 of the superficial 40% of weight bearing femoral cartilage after exercise. These in vivo observations agree well with published ex vivo results and support the hypothesis that cartilage compression results in greater anisotropy of superficial collagen fibers. PMID- 15550377 TI - Abca7 null mice retain normal macrophage phosphatidylcholine and cholesterol efflux activity despite alterations in adipose mass and serum cholesterol levels. AB - Mutations in the A class of ATP-binding cassette transporters (ABCA) are causally implicated in three human diseases: Tangier disease (ABCA1), Stargadt's macular degeneration (ABCA4), and neonatal respiratory failure (ABCA3). Both ABCA1 and ABCA4 have been shown to transport lipids across cellular membranes, and ABCA3 may play a similar role in transporting pulmonary surfactant. Although the functions of the other 10 ABCA class transporters identified in the human genome remain obscure, ABCA7-transfected cells have been shown to efflux lipids in response to stimulation by apolipoprotein A-I. In an effort to elucidate the physiologic role of ABCA7, we generated mice lacking this transporter (Abca7-/- mice). Homozygous null mice were produced from intercrosses of heterozygous null mice at the expected Mendelian frequency and developed normally without any obvious phenotypic abnormalities. Cholesterol and phospholipid efflux stimulated by apolipoprotein A-I from macrophages isolated from wild type and Abca7-/- mice did not differ, suggesting that these activities may not be central to the physiological role of the transporter in vivo. Abca7-/- females, but not males, had significantly less visceral fat and lower total serum and high density lipoprotein cholesterol levels than wild type, gender-matched littermates. ABCA7 expression was detected in hippocampal and cortical neurons by in situ hybridization and in brain and white adipose tissue by Western blotting. Induction of adipocyte differentiation from 3T3 fibroblasts in culture led to a marked increase in ABCA7 expression. These studies suggest that ABCA7 plays a novel role in lipid and fat metabolism that Abca7-/- mice can be used to elucidate. PMID- 15550378 TI - Epigenetic silencing of the human nucleotide excision repair gene, hHR23B, in interleukin-6-responsive multiple myeloma KAS-6/1 cells. AB - During tumorigenesis, selective proliferative advantage in certain cell subsets is associated with accumulation of multiple genetic alterations. For instance, multiple myeloma is characterized by frequent karyotypic instability at the earliest stage, progressing to extreme genetic abnormalities as the disease progresses. These successive genetic alterations can be attributed, in part, to defects in DNA repair pathways, perhaps based on epigenetic gene silencing of proteins involved in DNA damage repair. Here we report epigenetic hypermethylation of the hHR23B gene, a key component of the nucleotide excision repair in response to DNA damage, in interleukin-6 (IL-6)-responsive myeloma KAS 6/1 cells. This hypermethylation was significantly abated by Zebularine, a potent demethylating agent, with a consequent increase in the hHR23B mRNA level. Subsequent removal of this drug and supplementation with IL-6 in the culture medium re-established DNA hypermethylation of the hHR23B gene and silencing of mRNA expression levels. The inclination of DNA to be remethylated, at least within the hHR23B gene promoter region, reflects an epigenetic driving force by the cytogenetic/tumorigenic status of KAS-6/1 myeloma. The IL-6 response of KAS 6/1 myeloma also raises a question of whether the proneoplastic growth factor, such as IL-6, supports the epigenetic silencing of important DNA repair genes via promoter hypermethylation during the development of multiple myeloma. PMID- 15550379 TI - Mechanistic insights into the suppression of drug resistance by human immunodeficiency virus type 1 reverse transcriptase using alpha-boranophosphate nucleoside analogs. AB - A class of amino acid substitutions in drug-resistant HIV-1 reverse transcriptase (RT) is responsible for the selectively impaired incorporation of the nucleotide analog inhibitor into DNA. We have shown previously that alpha-boranophosphate nucleoside analogs suppress RT-mediated resistance when the catalytic rate is responsible for drug resistance such as in the case of K65R and dideoxy (dd)NTPs, and Q151M toward AZTTP and ddNTPs. Here, we extend this property to BH3-d4TTP and BH3-3TCTP toward their clinically relevant mutants Q151M and M184V, respectively. Pre-steady-state kinetics on mutants of the Q151M RT family reveal a 3-5-fold resistance to d4TTP. This resistance is suppressed using BH3-d4TTP. Likewise, resistance to 3TCTP by M184V RT (30-fold) and K65R/M184V RT (180-fold) is suppressed using BH3-3TCTP because of a 160-fold acceleration of the catalytic constant kpol. Mechanistic insights into the rate enhancement were obtained using various alpha-boranophosphate nucleotides. The presence of the BH3 group renders kpol independent of amino acid substitutions present in RT. Indeed, the approximately 100-fold decrease in polymerase activity caused by the R72A substitution is restored to wild-type levels using BH3-dTTP. Metal ion titration studies show that alpha-boranophosphate nucleoside analogs enhance 3-8-fold the binding of Mg2+ ions to the active site of the RT.DNA.dNTP complex and alleviate the requirement of critical amino acids involved in phosphodiester bond formation. To our knowledge, this is the first example of rescue of polymerase activity by means of a nucleotide analog. PMID- 15550380 TI - Insulin-like growth factor-binding protein-3 plays an important role in regulating pharyngeal skeleton and inner ear formation and differentiation. AB - Insulin-like growth factor-binding protein (IGFBP)-3 is the major insulin-like growth factor (IGF) carrier protein in the bloodstream. IGFBP-3 prolongs the half life of circulating IGFs and prevents their potential hypoglycemic effect. IGFBP 3 is also expressed in many peripheral tissues in fetal and adult stages. In vitro, IGFBP-3 can inhibit or potentiate IGF actions and even possesses IGF independent activities, suggesting that local IGFBP-3 may also have paracrine/autocrine function(s). The in vivo function of IGFBP-3, however, is unclear. In this study, we elucidate the developmental role of IGFBP-3 using the zebrafish model. IGFBP-3 mRNA expression is first detected in the migrating cranial neural crest cells and subsequently in pharyngeal arches in zebrafish embryos. IGFBP-3 mRNA is also persistently expressed in the developing inner ears. To determine the role of IGFBP-3 in these tissues, we ablated the IGFBP-3 gene product using morpholino-modified antisense oligonucleotides (MOs). The IGFBP-3 knocked down embryos had delayed pharyngeal skeleton morphogenesis and greatly reduced pharyngeal cartilage differentiation. Knockdown of IGFBP-3 also significantly decreased inner ear size and disrupted hair cell differentiation and semicircular canal formation. Furthermore, reintroduction of a MO-resistant form of IGFBP-3 "rescued" the MO-induced defects. These findings suggest that IGFBP-3 plays an important role in regulating pharyngeal cartilage and inner ear development and growth in zebrafish. PMID- 15550381 TI - Activation domains from both monomers contribute to transcriptional stimulation by sterol regulatory element-binding protein dimers. AB - Sterol regulatory element-binding proteins (SREBPs) are basic helix-loop-helix leucine zipper proteins that act as dimers to activate genes in lipid metabolism. Three SREBP isoforms, 1a, 1c, and 2, are expressed at varying levels in different tissues. Thus, homo- and heterodimers probably contribute to overall SREBP activity. No studies have directly evaluated the formation or activation properties of SREBP homo- and heterodimers. Studies with overexpressed SREBP monomers are inconclusive regarding the function of a particular SREBP dimer because of potential dimerization with endogenous proteins. To assess activation by a particular SREBP dimer, we fused DNA encoding individual monomers together via a predicted flexible polypeptide tether. Tethered SREBP dimers bound DNA equivalently to the monomeric proteins and were resistant to dominant negative SREBP-1 inhibition, confirming preferential formation of intramolecular dimers. Tethered SREBP-1a and -2 homodimers, similar to the monomeric forms, activated target genes more robustly than tethered SREBP-1c homodimers. A forced SREBP-1a/2 heterodimer had similar activity to the respective homodimers. However, SREBP-1c in a heterodimer with either SREBP-1a or -2 attenuated the activity relative to the SREBP-1a or -2 homodimers. These experiments provide some of the first data showing that the integrity of both activation domains in a dimeric transcription factor is required for maximal activity. In addition, the results support a model where changes in SREBP-1c protein expression that occur in response to insulin signaling and liver X receptor signaling would be predicted to increase or decrease overall SREBP activity in a tissue-specific fashion depending on the initial fractional contribution of SREBP-1c to total cellular levels of SREBP. PMID- 15550382 TI - NMR structural comparison of the cytoplasmic juxtamembrane domains of G-protein coupled CB1 and CB2 receptors in membrane mimetic dodecylphosphocholine micelles. AB - The fourth cytoplasmic domain, the so-called C-terminal juxtamembrane segment or helix VIII, has been identified in numerous G-protein-coupled receptors and exhibits unique functional characteristics. Efforts have been devoted to studying the juxtamembrane segment in order to understand the biological importance of the segment in G-protein activation of the cannabinoid CB1 and CB2 receptors. Recent biochemical data revealed that the CB1 C-terminal juxtamembrane peptide fragment CB1-(401-417) can directly activate the G-protein and also showed that the specificity of the signal transduction activation by the C-terminal juxtamembrane region is unique to the CB1 receptor but not to the CB2 receptor (Mukhopadhyay, S., and Howlett, A. C. (2001) Eur. J. Biochem. 268, 499-505). However, there is experimental work, not yet reported, on the conformational analyses and structural comparison between the respective helix VIII segments of the two receptors. In the present study, we have examined the conformational specificities of the cytoplasmic helical domains for both cannabinoid receptors. Three-dimensional structural features of two synthetic CB1 and CB2 peptides, CB1I397-G418 and CB2I298-K319, respectively, in membrane mimetic DPC micelles were studied using a combined high resolution NMR and computer modeling approach. Comparisons of the NMR-determined structures of the two peptides as well as their correspondent mutant peptides revealed their conformational properties and salt bridge dissimilarity, which might help us to understand the different structural roles of the fourth cytoplasmic helices in the function and regulation of CB1 and CB2 receptors. PMID- 15550383 TI - Evidence for a role of the exocyst in insulin-stimulated Glut4 trafficking in 3T3 L1 adipocytes. AB - Insulin stimulates glucose transport in adipocytes and muscle by inducing the redistribution of Glut4 from intracellular locations to the plasma membrane. The fusion of Glut4-containing vesicles at the plasma membrane is known to involve the target SNAREs syntaxin 4 and SNAP-23 and the vesicle SNARE VAMP2. Little is known about the initial docking of Glut4 vesicles with the plasma membrane. A recent report has implicated Exo70, a component of the mammalian exocyst complex, in the initial interaction of Glut4 vesicles with the adipocyte plasma membrane. Here, we have examined the role of two other exocyst components, rsec6 and rsec8. We show that insulin promotes a redistribution of rsec6 and rsec8 to the plasma membrane and to cytoskeletal fractions within 3T3-L1 adipocytes but does not modulate levels of these proteins co-localized with Glut4. We further show that adenoviral-mediated overexpression of either rsec6 or rsec8 increases the magnitude of insulin-stimulated glucose transport in 3T3-L1 adipocytes. By contrast, overexpression of rsec6 or rsec8 did not increase the extent of the secretion of adipsin or ACRP30 from adipocytes and had no discernible effect on transferrin receptor traffic. Collectively, our data support a role for the exocyst in insulin-stimulated glucose transport and suggest a model by which insulin-dependent relocation of the exocyst to the plasma membrane may contribute to the specificity of Glut4 vesicle docking and fusion with the adipocyte plasma membrane. PMID- 15550384 TI - Iron-mediated H2O2 production as a mechanism for cell type-specific inhibition of tumor necrosis factor alpha-induced but not interleukin-1beta-induced IkappaB kinase complex/nuclear factor-kappaB activation. AB - Coordinated and specific regulation of tumor necrosis factor (TNF) and interleukin (IL)-1 signaling pathways and how and whether they are modified by different agents are key events for proper immune responses. The IkappaB kinase complex (IKK)/NF-kappaB and JNK/AP-1 pathways are central mediators of TNF and IL 1 during inflammatory responses. Here we show that l-mimosine, a toxic non protein amino acid that has been shown to reduce serum TNFalpha levels and affect inflammatory responses, specifically inhibits TNF-induced IKK but not JNK in a cell type-specific manner. l-Mimosine did not affect IKK and NF-kappaB activation by IL-1beta. l-Mimosine caused cell cycle arrest at G(1)-S phase, but inhibition of IKK was found to be independent of cell cycle arrest. Treatment of cells with l-mimosine resulted in production of H(2)O(2). Addition of FeSO(4) restored IKK activation by TNFalpha as did ectopic expression of catalase or pretreatment of cells with N-aceltyl-l-cysteine, indicating a role for intracellular H(2)O(2) as a mediator of inhibition. Cleavage and degradation of TNF pathway components TNFR1, RIP, and Hsp90 were observed in l-mimosine and H(2)O(2) treated cells indicating a putative mechanism for selective inhibition of TNF but not IL-1beta induced IKK activation. PMID- 15550385 TI - Novel parathyroid hormone (PTH) antagonists that bind to the juxtamembrane portion of the PTH/PTH-related protein receptor. AB - Current antagonists for the parathyroid hormone (PTH)/PTH-related protein (PTHrP) receptor (PTHR) are N-terminally truncated or N-terminally modified analogs of PTH(1-34) or PTHrP(1-34) and are thought to bind predominantly to the N-terminal extracellular (N) domain of the receptor. We hypothesized that ligands that bind only to PTHR region comprised of the extracellular loops and seven transmembrane helices (the juxtamembrane or J domain) could also antagonize the PTHR. To test this, we started with the J domain-selective agonists [Gln(10),Ala(12),Har(11),Trp(14),Arg(19) (M)]PTH(1-21), [M]PTH(1-15), and [M]PTH(1-14), and introduced substitutions at positions 1-3 that were predicted to dissociate PTHR binding and cAMP signaling activities. Strong dissociation was observed with the tri-residue sequence diethylglycine (Deg)(1)-para-benzoyl-l phenylalanine (Bpa)(2)-Deg(3). In HKRK-B7 cells, which express the cloned human PTHR, [Deg(1,3),Bpa(2),M]PTH(1-21), [Deg(1,3),Bpa(2),M]PTH(1-15), and [Deg(1,3),Bpa(2),M]PTH(1-14) fully inhibited (IC(50)s = 100-700 nm) the binding of (125)I-[alpha-aminoisobutyric acid(1,3),M]PTH(1-15) and were severely defective for stimulating cAMP accumulation. In ROS 17/2.8 cells, which express the native rat PTHR, [Deg(1,3),Bpa(2),M]PTH(1-21) and [Deg(1,3),Bpa(2),M]PTH(1 15) antagonized the cAMP-agonist action of PTH(1-34), as did PTHrP(5-36) (IC(50)s = 0.7 microm, 2.6 microm, and 36 nm, respectively). In COS-7 cells expressing PTHR-delNt, which lacks the N domain of the receptor, [Deg(1,3),Bpa(2), M]PTH(1 21) and [Deg(1,3),Bpa(2),M]PTH(1-15) inhibited the agonist actions of [alpha aminoisobutyric acid(1,3)]PTH(1-34) and [M]PTH(1-14) (IC(50)s approximately 1 microm), whereas PTHrP(5-36) failed to inhibit. [Deg(1,3),Bpa(2),M]PTH(1-14) inhibited the constitutive cAMP-signaling activity of PTHR-tether-PTH(1-9), in which the PTH(1-9) sequence is covalently linked to the PTHR J domain, as well as that of PTHR(cam)H223R. Thus, the J-domain-selective N-terminal PTH fragment analogs can function as antagonists as well as inverse agonists for the PTHR. The new ligands described should be useful for further studies of the ligand binding and activation mechanisms that operate in the critical PTHR J domain. PMID- 15550386 TI - Insulin-like growth factor-1 (IGF-1) inversely regulates atrophy-induced genes via the phosphatidylinositol 3-kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway. AB - Skeletal muscle size is regulated by anabolic (hypertrophic) and catabolic (atrophic) processes. We first characterized molecular markers of both hypertrophy and atrophy and identified a small subset of genes that are inversely regulated in these two settings (e.g. up-regulated by an inducer of hypertrophy, insulin-like growth factor-1 (IGF-1), and down-regulated by a mediator of atrophy, dexamethasone). The genes identified as being inversely regulated by atrophy, as opposed to hypertrophy, include the E3 ubiquitin ligase MAFbx (also known as atrogin-1). We next sought to investigate the mechanism by which IGF-1 inversely regulates these markers, and found that the phosphatidylinositol 3 kinase/Akt/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway, which we had previously characterized as being critical for hypertrophy, is also required to be active in order for IGF-1-mediated transcriptional changes to occur. We had recently demonstrated that the IGF1/PI3K/Akt pathway can block dexamethasone induced up-regulation of the atrophy-induced ubiquitin ligases MuRF1 and MAFbx by blocking nuclear translocation of a FOXO transcription factor. In the current study we demonstrate that an additional step of IGF1 transcriptional regulation occurs downstream of mTOR, which is independent of FOXO. Thus both the Akt/FOXO and the Akt/mTOR pathways are required for the transcriptional changes induced by IGF-1. PMID- 15550387 TI - Regulation of chicken ccn2 gene by interaction between RNA cis-element and putative trans-factor during differentiation of chondrocytes. AB - CCN2/CTGF is a multifunctional growth factor. Our previous studies have revealed that CCN2 plays important roles in both growth and differentiation of chondrocytes and that the 3'-untranslated region (3'-UTR) of ccn2 mRNA contains a cis-repressive element of gene expression. In the present study, we found that the stability of chicken ccn2 mRNA is regulated in a differentiation stage dependent manner in chondrocytes. We also found that stimulation by bone morphogenetic protein 2, platelet-derived growth factor, and CCN2 stabilized ccn2 mRNA in proliferating chondrocytes but that it destabilized the mRNA in prehypertrophic-hypertrophic chondrocytes. The results of a reporter gene assay revealed that the minimal repressive cis-element of the 3'-UTR of chicken ccn2 mRNA was located within the area between 100 and 150 bases from the polyadenylation tail. Moreover, the stability of ccn2 mRNA was correlated with the interaction between this cis-element and a putative 40-kDa trans-factor in nuclei and cytoplasm. In fact, the binding between them was prominent in proliferating chondrocytes and attenuated in (pre)hypertrophic chondrocytes. Stimulation by the growth factors repressed the binding in proliferating chondrocytes; however, it enhanced it in (pre)hypertrophic chondrocytes. Therefore, gene expression of ccn2 mRNA during endochondral ossification is properly regulated, at least in part, by changing the stability of the mRNA, which arises from the interaction between the RNA cis-element and putative trans factor. PMID- 15550388 TI - Expression of DGAT2 in white adipose tissue is regulated by central leptin action. AB - Acyl-CoA:diacylglycerol acyltransferase (DGAT) enzymes catalyze the final step in mammalian triglyceride synthesis, and their functions are considered to be involved in the mechanisms of obesity, insulin resistance, and leptin resistance. Insulin receptor substrate-2 (IRS-2)-deficient mice exhibit obesity-associated with hypertrophic adipocytes and leptin resistance. Screening for transcripts of genes involved in fatty acid and triglyceride synthesis to investigate the mechanism of the hypertrophic change in the adipocytes showed that expression of DGAT2 mRNA was up-regulated in the white adipose tissue (WAT) of Irs2-/- mice, whereas that of DGAT1 was down-regulated. This reciprocal expression of DGAT1 and DGAT2 was also observed in WAT of leptin-deficient ob/ob mice. A high fat diet also resulted in increased DGAT2 and reduced DGAT1 in the WAT of C57BL/6 mice. Induction of adipocyte hypertrophy in vitro up-regulated both DGAT1 and DGAT2 expression in 3T3-L1 cells, suggesting that adipocyte non-autonomous mechanism in vivo is required for the reciprocal changes in expression of DGAT1 and DGAT2. In fact, intracerebroventricular infusion of leptin reduced DGAT2 expression in WAT of Irs2-/- mice and ob/ob mice, independently of DGAT1 expression. We propose the hypothesis that leptin regulates adipocyte size by altering expression patterns of DGAT via central nervous system to determine the levels of triglyceride synthesis. PMID- 15550389 TI - Cystic renal neoplasia following conditional inactivation of apc in mouse renal tubular epithelium. AB - Alterations in Wnt/beta-catenin signaling have been linked to abnormal kidney development and tumorigenesis. To gain more insights into the effects of these alterations, we created mice carrying a conditional deletion of the Apc tumor suppressor gene specifically in the renal epithelium. As expected, the loss of Apc leads to increased levels of beta-catenin protein in renal epithelium. Most of these mice die shortly after birth, and multiple kidney cysts were found upon histological examination. Only rarely did these animals survive to adulthood. Analysis of these adults revealed severely cystic kidneys associated with the presence of renal adenomas. Our results confirm an important role for proper regulation of Wnt/beta-catenin signaling in renal development and provide evidence that dysregulation of the pathway can initiate tumorigenesis in the kidney. PMID- 15550390 TI - Interferon-gamma down-regulates adenosine 2b receptor-mediated signaling and short circuit current in the intestinal epithelia by inhibiting the expression of adenylate cyclase. AB - Adenosine is an endogenous signaling molecule that is highly up-regulated in inflammatory states. Adenosine acts through the A2b receptor, a G protein-coupled receptor that couples positively to Galpha(s) and activates adenylate cyclase. This leads to cAMP-mediated electrogenic chloride secretion in intestinal epithelia. To better understand the regulation of the A2b receptor in intestinal epithelia, we studied the effects of interferon-gamma (IFN-gamma), a potent immunomodulatory cytokine, in the T84 cell line. Pretreatment of cells with 500 units/ml IFN-gamma for 12 h inhibited an adenosine-induced short circuit current (Isc) without affecting the transepithelial resistance. Under these conditions, IFN-gamma did not inhibit the protein expression or membrane recruitment of the A2b receptor, shown to be essential for its function. Interestingly, IFN-gamma inhibited cAMP levels as well as its downstream signaling pathway as shown by the inhibition of adenosine-induced phosphorylation of cAMP response element-binding protein and protein kinase A activity. Similar studies with forskolin, a direct activator of adenylate cyclase, also demonstrated inhibition of cAMP and its downstream response by IFN-gamma. However, IFN-gamma did not affect secretory responses to the calcium-dependent secretagogue carbachol or cAMP analog 8-bromo cAMP, indicating that normal secretory responses to adequate second messengers in IFN-gamma-treated cells are achievable. Moreover, IFN-gamma inhibited the expression of adenylate cyclase isoforms 5 and 7. In conclusion, we demonstrate that IFN-gamma down-regulates adenosine-mediated signaling possibly through the direct inhibition of adenylate cyclase expression. We propose that IFN-gamma may acutely affect global cAMP-mediated responses in the intestinal epithelia, thereby decreasing secretory responses, which may consequently aggravate inflammatory processes. PMID- 15550391 TI - Peptidase activity of the Escherichia coli Hsp31 chaperone. AB - Hsp31, the Escherichia coli hcha gene product, is a molecular chaperone whose activity is inhibited by ATP at high temperature. Its crystal structure reveals a putative Cys(184), His(185), and Asp(213) catalytic triad similar to that of the Pyrococcus horikoshii protease PH1704, suggesting that it should display a proteolytic activity. A preliminary report has shown that Hsp31 has an exceedingly weak proteolytic activity toward bovine serum albumin and a peptidase activity toward two peptide substrates with small amino acids at their N terminus (alanine or glycine), but the physiological significance of this observation remains unclear. In this study, we report that Hsp31 does not diplay any significant proteolytic activity but has peptidolytic activity. The aminopeptidase cleavage preference of Hsp31 is Ala > Lys > Arg > His, suggesting that Hsp31 is an aminopeptidase of broad specificity. Its aminopeptidase activity is inhibited by the thiol reagent iodoacetamide and is completely abolished in a C185A mutant, which is consistent with Hsp31 being a cysteine peptidase. The aminopeptidase activity of Hsp31 is also inhibited by EDTA and 1,10 phenanthroline, in concordance with the importance of the putative His(85), His(122), and Glu(90) metal-binding site revealed by crystallographic studies. An Hsp31-deficient mutant accumulates more 8-12-mer peptides than its parental strain, and purified Hsp31 can transform these peptides into smaller peptides, suggesting that Hsp31 has an important peptidase function both in vivo and in vitro. Proteins interacting with Hsp31 have been identified by reverse purification of a crude E. coli extract on an Hsp31-affinity column, followed by SDS-polyacrylamide electrophoresis and mass spectrometry. The ClpA component of the ClpAP protease, the chaperone GroEL, elongation factor EF-Tu, and tryptophanase were all found to interact with Hsp31, thus substantiating the role of Hsp31 as both chaperone and peptidase. PMID- 15550392 TI - Macromolecular accessibility of fluorescent taxoids bound at a paclitaxel binding site in the microtubule surface. AB - The macromolecular accessibility of the paclitaxel binding site in microtubules has been investigated using a fluorescent taxoid and antibodies against fluorescein, which cannot diffuse into the microtubule lumen. The formation of a specific ternary complex of microtubules, Hexaflutax (7-O-{N-[6-(fluorescein-4' carboxamido)-n-hexanoyl]-l-alanyl}paclitaxel) and 4-4-20 IgG (a monoclonal antibody against fluorescein) has been observed by means of sedimentation and electron microscopy methods. The kinetics of binding of the antibody to microtubule-bound Hexaflutax has been measured. The quenching of the observed fluorescence is fast (k+ 2.26 +/- 0.25 x 10(6) m(-1) s(-1) at 37 degrees C), indicating that the fluorescein groups of Hexaflutax are exposed to the outer solvent. The velocity of the reaction is linearly dependent on the antibody concentration, indicating that a bimolecular reaction is being observed. Another fluorescent taxoid (Flutax-2) bound to microtubules has also been shown to be rapidly accessible to polyclonal antibodies directed against fluorescein. A reduced rate of Hexaflutax quenching by the antibody is observed in microtubule associated proteins containing microtubules or in native cellular cytoskeletons. It can be concluded that the fluorescent taxoids bind to an outer site on the microtubules that is shared with paclitaxel. Paclitaxel would be internalized in a further step of binding to reach the known luminal site, this step being blocked in the case of the fluorescent taxoids. Because the fluorescent ligands are able to induce microtubule assembly, binding to the outer site should be enough to induce assembly by a preferential binding mechanism. PMID- 15550393 TI - Activation of p21-activated kinase 6 by MAP kinase kinase 6 and p38 MAP kinase. AB - The p21-activated kinases (PAKs) contain an N-terminal Cdc42/Rac interactive binding domain, which in the group 1 PAKs (PAK1, 2, and 3) regulates the activity of an adjacent conserved autoinhibitory domain. In contrast, the group 2 PAKs (PAK4, 5, and 6) lack this autoinhibitory domain and are not activated by Cdc42/Rac binding, and the mechanisms that regulate their kinase activity have been unclear. This study found that basal PAK6 kinase activity was repressed by a p38 mitogen-activated protein (MAP) kinase antagonist and could be strongly stimulated by constitutively active MAP kinase kinase 6 (MKK6), an upstream activator of p38 MAP kinases. Mutation of a consensus p38 MAP kinase target site at serine 165 decreased PAK6 kinase activity. Moreover, PAK6 was directly activated by MKK6, and mutation of tyrosine 566 in a consensus MKK6 site (threonine-proline-tyrosine, TPY) in the activation loop of the PAK6 kinase domain prevented activation by MKK6. PAK6 activation by MKK6 was also blocked by mutation of an autophosphorylated serine (serine 560) in the PAK6 activation loop, indicating that phosphorylation of this site is necessary for MKK6-mediated activation. PAK4 and PAK5 were similarly activated by MKK6, consistent with a conserved TPY motif in their activation domains. The activation of PAK6 by both p38 MAP kinase and MKK6 suggests that PAK6 plays a role in the cellular response to stress-related signals. PMID- 15550394 TI - Site-specific disulfide capture of agonist and antagonist peptides on the C5a receptor. AB - The manner by which peptidic ligands bind and activate their corresponding G protein-coupled receptors is not well understood. One of the better characterized peptidic ligands is the chemotactic cytokine complement factor 5a (C5a), a 74 amino acid helical bundle. Previous studies showed 6-mer peptide analogs derived from the C terminus of the C5a ligand can bind to C5aR (Kd values approximately 0.1-1 microm) and either agonize or antagonize the receptor (Gerber, B. O., Meng, E. C., Dotsch, V., Baranski, T. J., and Bourne, H. R. (2001) J. Biol. Chem. 276, 3394-3400). Here, we provide direct biochemical data using disulfide trapping to support a model that these peptides bind within a transmembrane helical triad formed by alpha-helices III, VI, and VII. We show that the three amino acids on the C terminus of the peptide analogs bind too weakly to exert a functional effect themselves. However, when a cysteine residue is placed on their N terminus they can be trapped by disulfide interchange to specific cysteines in helix III and VI and not to other cysteines, engineered into the C5aR. The trapped peptides function as agonists or partial antagonists, similar to the non-covalent parents from which they were derived. These data help to further refine the binding mode for C5a to the C5aR and suggest an approach and a binding site that may be applicable to studying other peptide binding receptors. PMID- 15550395 TI - Separation force measurements reveal different types of modulation of E-cadherin based adhesion by nectin-1 and -3. AB - Nectins are Ca2+-independent cell adhesion molecules found at cadherin-based adherens junctions. We used a dual pipette assay that measures the forces required to separate cell doublets to determine how nectins affect the formation and strength of cell-cell adhesion. Less force was required to separate doublets of L cells expressing nectin-1 or nectin-3 than to separate doublets of E cadherin-expressing cells. Heterodimers formed between cells expressing nectin-1 or nectin-3 adhered more strongly than homodimers. Nectin-3 that does not trans interact with nectin-1 inhibited E-cadherin-mediated adhesion. However, the extracellular fragment of nectin-1 did not have an agonistic effect on E-cadherin dependent cell adhesion when it trans-interacted with nectin-3, expressed at high levels in cells. In contrast, the extracellular fragment of nectin-3 had a significant agonistic effect on cadherin-based adhesion when it interacted with endogenous nectin-1, expressed at low levels in cells. Our results indicate that E-cadherin is the key molecule involved in cell adhesion and that the regulation of E-cadherin-based adhesion involving cellular nectin-1 trans-interacting with nectin-3 is qualitatively different from that involving cellular nectin-3 trans interacting with nectin-1 and depends on the nectin levels expressed by cells. PMID- 15550396 TI - Supramolecular amyloid-like assembly of the polypeptide sequence coded by exon 30 of human tropoelastin. AB - Elastin is known to self-aggregate in twisted-rope filaments. However, an ultrastructural organization different from the fibrils typical of elastin, but rather similar to those shown by amyloid networks, is shown by the polypeptide sequence encoded by exon 30 of human tropoelastin. To better understand the molecular properties of this sequence to give amyloid fibers, we used CD, NMR, and FTIR (Fourier transform infrared spectroscopy) to identify the structural characteristics of the peptide. In this study, we have demonstrated, by FTIR, that antiparallel beta-sheet conformation is predominant in the exon 30 fibers. These physical-chemical studies were combined with transmission electron microscopy and atomic force microscopy to analyze the supramolecular structure of the self-assembled aggregate. These studies show the presence of fibrils that interact side-by-side probably originating from an extensive self-interaction of elemental cross beta-structures. Similar sequences, of the general type XGGZG(X, Z = V, L, A, I), are widely found in many proteins such as collagens IV and XVII, major prion protein precursor, amyloid beta A4 precursor protein-binding family, etc., thus suggesting that this sequence could be involved in contributing to the self-assembly of amyloid fibers even in other proteins. PMID- 15550397 TI - GATA-6 can act as a positive or negative regulator of smooth muscle-specific gene expression. AB - The GATA-4/5/6 family of transcription factors is important for the development of the cardiovascular system and the visceral endoderm. GATA-6 is the only family member expressed in vascular smooth muscle cells and has been shown to be important for controlling the phenotype of these cells following vascular injury. To clarify further the role of GATA-6 in regulating vascular smooth muscle differentiation, we directly examined its ability to regulate the promoters of smooth muscle-specific genes. This analysis revealed that GATA-6 strongly repressed telokin promoter activity. In contrast, GATA-6 activated the smooth muscle myosin heavy chain and smooth muscle alpha-actin promoters and had no significant effect on the SM22alpha promoter. Gel mobility shift assays demonstrate that GATA-6 binds to a consensus site adjacent to the CArG box in the telokin promoter. GATA-6 did not interfere with the serum-response factor stimulated promoter activity but blocked myocardin-induced activation of the telokin promoter. In contrast, GATA-6 and myocardin resulted in synergistic activation of the smooth muscle myosin heavy chain promoter. Consistent with these findings, overexpression of GATA-6 in smooth muscle cells selectively inhibited expression of endogenous telokin, while simultaneously increasing expression of other smooth muscle proteins. These data suggest that GATA-6 selectively inhibits telokin expression by triggering the displacement of myocardin from the serum-response factor. As GATA-6 is expressed at high levels in vascular smooth muscle, this finding may explain the relatively low levels of telokin expression in the vascular system. These data also reveal a novel transcription regulatory mechanism by which GATA-6 can modulate the activity of the myocardin-serum-response factor complexes. PMID- 15550398 TI - Dystrophin Dp71 expression is down-regulated during myogenesis: role of Sp1 and Sp3 on the Dp71 promoter activity. AB - Dp71 expression is present in myoblasts but declines during myogenesis to avoid interfering with the function of dystrophin, the predominant Duchenne muscular dystrophy gene product in differentiated muscle fibers. To elucidate the transcriptional regulatory mechanisms operating on the developmentally regulated expression of Dp71, we analyzed the Dp71 expression and promoter activity during myogenesis of the C2C12 cells. We demonstrated that the cellular content of Dp71 transcript and protein decrease in myotubes as a consequence of the negative regulation that the differentiation stimulus exerts on the Dp71 promoter. Promoter deletion analysis showed that the 224-bp 5'-flanking region, which contains several Sp-binding sites (Sp-A to Sp-D), is responsible for the Dp71 promoter basal activity in myoblasts as well as for down-regulation of the promoter in differentiated cells. Electrophoretic mobility shift and chromatin immunoprecipitation assays indicated that Sp1 and Sp3 transcription factors specifically bind to the Sp-binding sites in the minimal Dp71 promoter region. Site-directed mutagenesis assay revealed that Sp-A is the most important binding site for the proximal Dp71 promoter activity. Additionally, cotransfection of the promoter construct with Sp1- and Sp3-expressing vectors into Drosophila SL2 cells, which lack endogenous Sp family, confirmed that these proteins activate specifically the minimal Dp71 promoter. Endogenous Sp1 and Sp3 proteins were detected only in myoblasts and not in myotubes, which indicates that the lack of these factors causes down-regulation of the Dp71 promoter activity in differentiated cells. In corroboration, efficient promoter activity was restored in differentiated muscle cells by exogenous expression of Sp1 and Sp3. PMID- 15550399 TI - Solution structure of prosurvival Mcl-1 and characterization of its binding by proapoptotic BH3-only ligands. AB - The B cell lymphoma-2 (Bcl-2) homologs myeloid cell leukemia-1 (Mcl-1) and A1 are prosurvival factors that selectively bind a subset of proapoptotic Bcl homology (BH) 3-only proteins. To investigate the molecular basis of the selectivity, we determined the solution structure of the C-terminal Bcl-2-like domain of Mcl-1. This domain shares features expected of a prosurvival Bcl-2 protein, having a helical fold centered on a core hydrophobic helix and a surface-exposed hydrophobic groove for binding its cognate partners. A number of residues in the binding groove differentiate Mcl-1 from its homologs, and in contrast to other Bcl-2 homologs, Mcl-1 has a binding groove in a conformation intermediate between the open structures characterized by peptide complexes and the closed state observed in unliganded structures. Mutagenesis of potential binding site residues was used to probe the contributions of groove residues to the binding properties of Mcl-1. Although mutations in Mcl-1 had little impact on binding, a single mutation in the BH3-only ligand Bad enabled it to bind both Mcl-1 and A1 while retaining its binding to Bcl-2, Bcl-xL, and Bcl-w. Elucidating the selective action of certain BH3-only ligands is required for delineating their mode of action and will aid the search for effective BH3-mimetic drugs. PMID- 15550400 TI - Cyclooxygenase-2 suppresses hypoxia-induced apoptosis via a combination of direct and indirect inhibition of p53 activity in a human prostate cancer cell line. AB - Although p53-inactivating mutations have been described in the majority of human cancers, their role in prostate cancer is controversial as mutations are uncommon, particularly in early lesions. p53 is activated by hypoxia and other stressors and is primarily regulated by the Mdm2 protein. Cyclooxygenase (COX)-2, an inducible enzyme that catalyzes the conversion of arachidonic acid to prostaglandins and other eicosanoids, is also induced by hypoxia. COX-2 and resultant prostaglandins increase tumor cell proliferation, resistance to apoptosis, and angiogenesis. Previous reports indicate a complex, reciprocal relationship between p53 and COX-2. To elucidate the effects of COX-2 on p53 in response to hypoxia, we transfected the COX-2 gene into the p53-positive, COX-2 negative MDA-PCa-2b human prostate cancer cell line. The expression of functional p53 and Mdm2 was compared in COX-2+ versus COX-2- cells under normoxic and hypoxic conditions. Our results demonstrated that hypoxia increases both COX-2 protein levels and p53 transcriptional activity in these cells. Forced expression of COX-2 increased tumor cell viability and decreased apoptosis in response to hypoxia. COX-2+ cells had increased Mdm2 phosphorylation in either normoxic or hypoxic conditions. Overexpression of COX-2 abrogated hypoxia-induced p53 phosphorylation and promoted the binding of p53 to Mdm2 protein in hypoxic cells. In addition, COX-2-expressing cells exhibited decreased hypoxia-induced nuclear accumulation of p53 protein. Finally, forced expression of COX-2 suppressed both basal and hypoxia-induced p53 transcriptional activity, and this effect was mimicked by the addition of PGE2 to wild-type cells. These results demonstrated a role for COX-2 in the suppression of hypoxia-induced p53 activity via both direct effects and indirect modulation of Mdm2 activity. These data imply that COX-2 positive prostate cancer cells can have impaired p53 function even in the presence of wild-type p53 and that p53 activity can be restored in these cells via inhibition of COX-2 activity. PMID- 15550401 TI - Parenteral metoclopramide for acute migraine: meta-analysis of randomised controlled trials. AB - OBJECTIVE: To assess the evidence from controlled trials on the efficacy and tolerability of parenteral metoclopramide for acute migraine in adults. DATA SOURCES: Cochrane Central Register of Controlled Trials, Medline, Embase, LILACS, CINAHL, conference proceedings, clinical practice guidelines, and other sources. SELECTION CRITERIA: Randomised controlled trials of parenteral metoclopramide for acute migraine in adults. RESULTS: We reviewed 596 potentially relevant abstracts and found 13 eligible trials totalling 655 adults. In studies comparing metoclopramide with placebo, metoclopramide was more likely to provide significant reduction in migraine pain (odds ratio 2.84, 95% confidence interval 1.05 to 7.68). Used as the only agent, metoclopramide showed mixed effectiveness when compared with other single agents. Heterogeneity of studies for combination treatment prevented statistical pooling. Treatments that did include metoclopramide were as, or more, effective than comparison treatments for pain, nausea, and relapse outcomes reported in all studies. CONCLUSIONS: Metoclopramide is an effective treatment for migraine headache and may be effective when combined with other treatments. Given its non-narcotic and antiemetic properties, metoclopramide should be considered a primary agent in the treatment of acute migraines in emergency departments. PMID- 15550402 TI - Non-specific effects of vaccination on child survival: prospective cohort study in Burkina Faso. AB - OBJECTIVE: To study the effect of vaccination on mortality before 2 years of age in a developing country. DESIGN: Prospective cohort study. SETTING: Rural communities in Burkina Faso. PARTICIPANTS: 9085 children born in the study area between 1985 and 1993. MAIN OUTCOME MEASURE: Child death rate. RESULTS: Mortality before 2 years of age was lower in children who had been vaccinated: those vaccinated with BCG only had significantly lower mortality (risk ratio for vaccinated v unvaccinated children 0.37, 95% confidence interval 0.29 to 0.48) as did those vaccinated with diphtheria, tetanus, and pertussis only (0.24, 0.13 to 0.43). The second dose of diphtheria, tetanus, and pertussis was not associated with lower mortality (0.80, 0.58 to 1.12). CONCLUSION: Vaccination with diphtheria, tetanus, and pertussis as well as BCG is associated with better survival of children up to 2 years of age. PMID- 15550403 TI - Legislation on smacking. PMID- 15550404 TI - Standardisation of glycated haemoglobin. PMID- 15550405 TI - Full engagement in health. PMID- 15550406 TI - Treating insomnia. PMID- 15550407 TI - Revisiting phenobarbital for epilepsy. PMID- 15550408 TI - Doctors angry as smoking ban is limited and delayed until 2008. PMID- 15550409 TI - Scotland to ban smoking in enclosed public places. PMID- 15550410 TI - Russia fails to ban drinking in public despite soaring alcoholism. PMID- 15550411 TI - UK government tightens rules for drug regulators. PMID- 15550412 TI - FDA will increase postmarketing surveillance of drugs. PMID- 15550413 TI - FDA bars own expert from evaluating risks of painkillers. PMID- 15550416 TI - Risk factors for death in infancy persist into older age groups in England and Wales. PMID- 15550417 TI - Coroner criticised over conduct of SARS inquest. PMID- 15550419 TI - Use of drug eluting stents increases in the US but only for some groups. PMID- 15550420 TI - Ranbaxy withdraws all its AIDS drugs from WHO list. PMID- 15550422 TI - Depression and obesity are major causes of maternal death in Britain. PMID- 15550424 TI - Authority consults public on paying women 1000 pounds sterling to donate eggs. PMID- 15550425 TI - Counting hospital activity: spells or episodes? PMID- 15550426 TI - The orthopaedic approach to managing osteoarthritis of the knee. PMID- 15550427 TI - Gastric rupture associated with use of the laryngeal mask airway during cardiopulmonary resuscitation. PMID- 15550428 TI - Feeding the preterm infant. PMID- 15550429 TI - Overcoming apathy in research on organophosphate poisoning. PMID- 15550430 TI - Is economic evaluation in touch with society's health values? PMID- 15550431 TI - Promoting walking and cycling as an alternative to using cars: perception of safety is biggest obstacle. PMID- 15550432 TI - Promoting walking and cycling as an alternative to using cars: comfort of cycling needs to be akin to that of using cars. PMID- 15550433 TI - Promoting walking and cycling as an alternative to using cars: vested interests doom puny healthcare interventions. PMID- 15550434 TI - How to deal with influenza: fever may be used as treatment. PMID- 15550435 TI - Headline about basal cell carcinoma was misleading. PMID- 15550436 TI - National service framework for children: framework shows a new vision of health, in life. PMID- 15550437 TI - National service framework for children: framework ignores children's growth. PMID- 15550438 TI - How to deal with influenza: worthwhile surveillance system is in action. PMID- 15550439 TI - Making sense of rising caesarean section rates: trials and speaking with patients take time. PMID- 15550440 TI - Making sense of rising caesarean section rates: reducing caesarean section rates should not be the primary objective. PMID- 15550441 TI - Making sense of rising caesarean section rates: caesarean section on demand is obstetric dilemma. PMID- 15550442 TI - Making sense of rising caesarean section rates: editorial made no sense of rising rates. PMID- 15550443 TI - Role of the endocannabinoid system in the development of tolerance to alcohol. AB - The present review evaluates the evidence that the endocannabinoid system plays in the development of tolerance to alcohol. The identification of a G-protein coupled receptor, namely, the cannabinoid receptor (CB(1) receptor), which was activated by Delta(9)-tetrahydrocannabinol (Delta(9)-THC), the major psychoactive component of marijuana, led to the discovery of endogenous cannabinoid agonists. Until now, four fatty acid derivatives identified to be arachidonylethanolamide (AEA), 2-arachidonylglycerol (2-AG), 2-arachidonylglycerol ether (noladin ether) and virodhamine have been isolated from both nervous and peripheral tissues. Both AEA and 2-AG have been shown to mimic the pharmacological and behavioural effects of Delta(9)-THC. The role of the endocannabinoid system in the development of tolerance to alcohol was not known until recently. Recent studies from our laboratory have implicated for the first time a role for the endocannabinoid system in development of tolerance to alcohol. Chronic alcohol treatment has been shown to down-regulate CB(1) receptors and its signal transduction. The observed downregulation of CB(1) receptor function results from the persistent stimulation of the receptors by AEA and 2-AG, the synthesis of which has been shown to be increased by chronic alcohol treatment. The enhanced formation of endocannabinoids may subsequently influence the release of neurotransmitters. It was found that the DBA/2 mice, known to avoid alcohol intake, have significantly reduced CB(1) receptor function in the brain, consistent with other studies in which the CB(1) receptor antagonist SR 141716A has been shown to block voluntary alcohol intake in rodents. Similarly, activation of the CB(1) receptor system promoted alcohol craving, suggesting a role for the CB(1) receptor gene in excessive alcohol drinking behaviour and development of alcoholism. Ongoing investigations may lead to a better understanding of the mechanisms underlying the development of tolerance to alcohol and to develop therapeutic strategies to treat alcoholism. PMID- 15550446 TI - Thiamine administration in alcohol-dependent patients. PMID- 15550444 TI - The endocannabinoid system: physiology and pharmacology. AB - The endogenous cannabinoid system is an ubiquitous lipid signalling system that appeared early in evolution and which has important regulatory functions throughout the body in all vertebrates. The main endocannabinoids (endogenous cannabis-like substances) are small molecules derived from arachidonic acid, anandamide (arachidonoylethanolamide) and 2-arachidonoylglycerol. They bind to a family of G-protein-coupled receptors, of which the cannabinoid CB(1) receptor is densely distributed in areas of the brain related to motor control, cognition, emotional responses, motivated behaviour and homeostasis. Outside the brain, the endocannabinoid system is one of the crucial modulators of the autonomic nervous system, the immune system and microcirculation. Endocannabinoids are released upon demand from lipid precursors in a receptor-dependent manner and serve as retrograde signalling messengers in GABAergic and glutamatergic synapses, as well as modulators of postsynaptic transmission, interacting with other neurotransmitters, including dopamine. Endocannabinoids are transported into cells by a specific uptake system and degraded by two well-characterized enzymes, the fatty acid amide hydrolase and the monoacylglycerol lipase. Recent pharmacological advances have led to the synthesis of cannabinoid receptor agonists and antagonists, anandamide uptake blockers and potent, selective inhibitors of endocannabinoid degradation. These new tools have enabled the study of the physiological roles played by the endocannabinoids and have opened up new strategies in the treatment of pain, obesity, neurological diseases including multiple sclerosis, emotional disturbances such as anxiety and other psychiatric disorders including drug addiction. Recent advances have specifically linked the endogenous cannabinoid system to alcoholism, and cannabinoid receptor antagonism now emerges as a promising therapeutic alternative for alcohol dependence and relapse. PMID- 15550447 TI - Attenuated salivary cortisol secretion under cue exposure is associated with early relapse. AB - AIMS: To test whether the risk of relapse in alcohol dependence is predicted by the subjective experience of cue exposure (CE) and/or cortisol reactivity to alcohol cues. METHODS: Salivary cortisol and self-ratings of 'tension' and 'desire to drink' were measured in 32 detoxified alcohol-dependent inpatients during CE sessions conducted in the first and third week of motivation enhancement therapy. Subjects completed the Toronto Alexithymia Scale (TAS-20) and the Abbreviated Alcohol Expectancy Questionnaire (B-AEQ) towards the end of the inpatient treatment to measure emotional self-awareness and the expected positive effects of alcohol. RESULTS: Six weeks after the end of the inpatient treatment, 15 patients were abstinent. Relapse was verified or was presumed for 17 patients. Those who had relapsed had shown an attenuated response to CE in the third week as an inpatient but did not differ from abstainers in terms of subjective reaction to cues. Subjective ratings of CE were not related to salivary cortisol or relapse but showed several associations with factors one and two of the TAS-20. The expectancy of enhanced social contacts by using alcohol (factor 1 of the B-AEQ) correlated negatively with the decline in salivary cortisol during the CE session in the third week of treatment. Subjective ratings of CE correlated with Alexithymiascores. CONCLUSIONS: Alcoholic patients who use alcohol to enhance their social contacts typically lack hypothalamo-hypophysical pituitary-adrenocortical (HPA) reactivity in the early period of abstention. They are at an increased risk of early relapse and perhaps use alcohol to increase cortisol secretion again. PMID- 15550448 TI - Effects of acetaldehyde and TNF alpha on the inhibitory kappa B-alpha protein and nuclear factor kappa B activation in hepatic stellate cells. AB - AIMS: Increased plasma tumour necrosis alpha (TNFalpha) and elevated monocyte nuclear factor kappa B (NF-kappaB) are associated with liver injury and inflammation in models of alcoholic liver disease and are found to be elevated in monocytes of patients with alcoholic hepatitis. Acetaldehyde enhances, whereas TNFalpha inhibits, transcription of the type I collagen promoters and type I collagen production. NF-kappaB, an inhibitor of the type I collagen promoters, is increased by both acetaldehyde and TNFalpha. This study determined the effects of acetaldehyde in comparison to the effects of TNFalpha on inhibitory kappa B-alpha (IkappaB-alpha) protein and NF-kappaB activation in hepatic stellate cells. METHODS: Activated rat hepatic stellate cells in culture were exposed to acetaldehyde or TNFalpha for short periods of time, following which the cells were harvested for the determination of IkappaB-alpha protein, IkappaB-alpha kinase activity and nuclear NF-kappaB. RESULTS: Acetaldehyde increased IkappaB alpha kinase activity and decreased IkappaB-alpha after 10 min of exposure, with recovery towards control levels at 20 min. In contrast, TNFalpha resulted in higher IkappaB-alpha kinase activity at 20 min than at 10 min, and similar low IkappaB-alpha at 10 and 20 min. Both acetaldehyde and TNFalpha enhanced nuclear NF-kappaB (p65), but acetaldehyde alone also increased NF-kappaB (p50). CONCLUSIONS: TNFalpha and acetaldehyde independently activate NF-kappaB by rapid enhancement of IkappaB-alpha kinase activity and degradation of IkB-alpha protein. Increased TNFalpha is the principal mechanism for the elevation of NF kappaB in severe alcoholic hepatitis. The elevation of NF-kappaB due to TNFalpha enhance liver injury, but inhibit fibrogenesis. In contrast, the effect of acetaldehyde in activating NF-kappaB is associated with increases in both liver injury and fibrogenesis, indicating that the effects of acetaldehyde on fibrogenesis are mediated by cytokines and by trans-acting factors other than NF kappaB. PMID- 15550449 TI - Ethanol induces higher BEC in CB1 cannabinoid receptor knockout mice while decreasing ethanol preference. AB - AIMS: Previous studies have shown that CB(1) cannabinoid receptors are involved in the behavioural effects induced by chronic ethanol administration in Wistar rats by using SR 141716, a CB(1) cannabinoid receptor antagonist. These studies have now been extended to investigate the effect of acute and chronic alcoholization on blood ethanol concentration (BEC) and ethanol preference in CB(1) knockout (-/-) mice. METHODS: BEC was monitored for a period of 8 h in both CB(1)(-/-) male mice and CB(1) male wild-type (+/+) mice, which had received an acute i.p. injection of ethanol in 1, 3 or 5 g/kg doses. Ethanol preference was assayed in both groups of male mice in non-forced ethanol administration and forced chronic pulmonary alcohol administration for 14 and 39 days, respectively. RESULTS: After an acute intraperitoneal ethanol injection of 5 g/kg, CB(1)(-/-) mice showed a significant higher BEC during the ethanol elimination stage than the CB(1)(+/+) mice. However, those in the 1 and 3 g/kg groups showed no significant difference. A 2-3 fold increase in BEC was observed in CB(1)(-/-) mice on days 10 and 11 after commencement of forced chronic pulmonary alcoholization in comparison with CB(1)(+/+) mice, although comparable BEC values were assayed in both groups on day 12. In addition, these CB(1)(-/-) mice showed a significantly lower preference for ethanol than CB(1)(+/+) mice. CONCLUSIONS: The studies on CB(1)(-/-) and CB(1)(+/+) mice have clearly confirmed the involvement of CB(1) receptor on ethanol induced behavioural effects and also revealed that CB(1) receptors may be implicated in ethanol absorption/distribution, particularly after administration of high ethanol doses. PMID- 15550450 TI - Delta9-THC reinstates beer- and sucrose-seeking behaviour in abstinent rats: comparison with midazolam, food deprivation and predator odour. AB - AIMS: Recent studies suggest that cannabinoid receptor agonists may promote relapse to drug-seeking behaviour after a period of abstinence. In this study, the ability of Delta(9)-tetrahydrocannabinol (THC) to reinstate previously reinforced responding for alcoholic and non-alcoholic beverages was assessed in rats using a novel lick-based paradigm. METHODS: Rats were initially given free access to beer (containing 4.5% ethanol v/v), near-beer (a beverage that looks and tastes like beer but contains <0.5% ethanol v/v) or isocaloric sucrose in their home cages for 3 weeks. They were then trained to lick at a tube to self administer the pre-exposed beverage in operant chambers under a VR10 schedule in 30-min sessions daily. After approximately 3 weeks of such access, the rats underwent an extinction procedure, so that licking at the tube produced no reward. Once responding had ceased, the rats were subjected to various reinstatement tests. RESULTS: In Experiment 1, the cannabinoid receptor agonist Delta(9)-THC (1 mg/kg) significantly reinstated responding, previously reinforced with beer or near-beer. The effect was unlikely to be caused by increased appetite because 24 h food-deprivation had no such effect. Exposure to cat odour in the test chamber failed to reinstate responding for beer or near-beer and caused a complete inhibition of responding. In Experiment 2, Delta(9)-THC (0.3 and 1 but not 3 mg/kg) again reinstated beer-seeking behaviour while the 1 mg/kg dose also reinstated responding in sucrose trained animals. Midazolam (0.15 mg/kg but not 0.5 or 1.5 mg/kg) produced a modest reinstatement of beer-seeking but had no effect on sucrose-seeking behaviour. CONCLUSIONS: The finding that Delta(9) THC can reinstate alcohol-seeking provides the impetus for further research into the involvement of the cannabinoid system in alcohol craving. However, the reinstatement of near-beer and sucrose-seeking behaviour caused by Delta(9)-THC suggests a relatively non-specific effect. This may perhaps be related to the stressor-like effects of cannabinoids, and their ability to activate key neural circuitry in the amygdala and bed nucleus of the stria terminalis. PMID- 15550451 TI - Interactions between cannabinoid and opioid receptor systems in the mediation of ethanol effects. AB - Over the past few years, advances in the investigation of the neurochemical circuits involved in the development and treatment of alcohol dependence have identified peptides and receptors as potential key targets in the treatment of problems related to alcohol consumption. The endogenous opioid system is modified by alcohol intake in areas of the brain related to reward systems, and differential basal levels of opioid gene expression are found in rodents with a high preference for ethanol. This suggests a greater vulnerability to alcohol consumption in relation to differences in genetic background. Further evidence of the involvement of opioid peptides in alcohol dependence is the ability of the opioid antagonist naltrexone to reduce alcohol intake in animal models of dependence and in alcohol-dependent patients. Abundant evidence indicates that the activation of cannabinoid receptors stimulates the release of opioid peptides, therefore the cannabinoid receptor antagonists may presumably alter opioid peptide release, thus facilitating the reduction of ethanol consumption. However, little is known about the effects of ethanol on the endogenous cannabinoid system, the vulnerability of cannabinoid receptors to alcohol intake or their neurochemical implications in reducing consumption of alcohol. In this paper, we review the role of opioid and cannabinoid receptor systems, their vulnerability to alcohol intake and the development of dependence, and the targeting of these systems in the treatment of alcoholism. PMID- 15550452 TI - The rare ERBB2 variant Ile654Val is associated with an increased familial breast cancer risk. AB - Overexpression of the proto-oncogene ERBB2 (HER2/NEU) has been observed in 20-30% of breast cancers involving poor prognosis. Genetic alterations within ERBB2 have been shown to induce carcinogenesis and metastasis. We investigated eight annotated single nucleotide polymorphisms for occurrence in familial breast cancer samples. The confirmed variants Ile654Val, Ile655Val and Ala1170Pro were analysed in subsequent epidemiological studies on familial breast cancer risk. While Ala1170Pro resides within a C-terminally located regulatory domain, the two adjacent polymorphisms Ile654Val and Ile655Val are part of the transmembrane domain. A case-control study analysing a cohort of 348 German familial breast cancer cases and 960 corresponding controls showed no significant association of either Ile655Val (OR = 1.05, 95% CI = 0.82-1.34, P = 0.728) or Ala1170Pro (OR = 0.94, 95% CI = 0.74-1.20, P = 0.632) with familial breast cancer risk. Differences in haplotype frequencies between cases and controls could also not be detected. The ERBB2 variant Ile654Val, however, revealed an increased risk for carriers of the heterozygous Val654 allele (OR = 2.56, 95% CI = 1.08-6.08, P = 0.028). The rare Val654 variant is linked with the more frequent Val655, resulting in two consecutive valine instead of two isoleucine residues within the transmembrane domain. Computational analyses suggest that the Val654-Val655 allele provokes receptor dimerization and activation, thus stimulating kinase activity and cell transformation. We hypothesize that ERBB2 Val654 represents an oncogenic variant which might, in addition, influence clinical outcome and predict a worse prognosis. PMID- 15550453 TI - Colorectal adenoma risk is modified by the interplay between polymorphisms in arachidonic acid pathway genes and fish consumption. AB - Associations between polymorphisms in genes (SNPs) involved in the arachidonic acid (AA) pathway and colorectal adenomas have been investigated in a Dutch case control study including 384 cases and 403 polyp-free controls. Twenty-one polymorphisms in seven candidate genes were studied and a potential modifying effect of fish consumption was considered. A protective effect on colorectal adenomas was found for the CT genotype of SNP H477H in PPARgamma and the GC genotype of SNP V102V in COX-2 (OR 0.63, 95% CI 0.45-0.89 and OR 0.65, 95% CI 0.46-0.92, respectively) compared with the homozygous major genotypes. An increase in adenoma risk was observed for the TC genotype of SNP c.2242T-->C in COX-2 (OR 1.47, 95% CI 1.07-2.00) compared with the TT genotype. Analysis with estimated haplotypes confirmed these associations and revealed three additional associations with COX-2, sPLA(2) and 15LOX haplotypes. Fish consumption modified the associations with COX-2 and PPARdelta genotypes. For SNP c.-789C-->T in PPARdelta the major genotype showed a decrease in adenoma risk for those in the highest tertile of fish consumption (T3), as compared with the lowest tertile (T1) (OR 0.65, 95% CI 0.41-1.02). Protective effects were also observed for SNPs V102V and c.2242T-->C in COX-2 and high fish intake. The interaction between fish consumption and c.2242T-->C was statistically significant, with an OR for the TT genotype and high fish consumption of 0.52 (95% CI 0.27-1.01) as compared with low fish intake. These results indicate that SNPs in genes involved in the AA pathway are associated with colorectal adenoma risk. Some of these associations are modified by fish consumption. PMID- 15550454 TI - Single nucleotide polymorphisms in the EXO1 gene and risk of colorectal cancer in a Japanese population. AB - EXO1 is a member of the RAD2 nuclease family and functions in DNA replication, repair and recombination. We investigated the relationship of single nucleotide polymorphisms (SNPs) at exon 10 (T439M) and exon 13 (P757L) of the EXO1 gene with development, progression and metastasis of colorectal cancer. For T439M, the Thr/Met genotype [odds ratio (OR) = 2.03, 95% confidence interval (CI) 1.04-3.98] and Thr/Met and Met/Met genotypes combined (OR = 2.37, 95% CI 1.23-4.56) demonstrated significant association with the development of colorectal cancer after adjusting for age, gender and smoking status. For P757L, patients with the Leu/Leu genotype showed a reduced risk of colorectal cancer (adjusted OR = 0.398, 95% CI 0.183-0.866) when the Pro/Leu and Pro/Pro genotypes were combined and used as the reference. The Leu/Leu genotype also had a reduced risk (adjusted OR = 0.373, 95% CI 0.164-0.850) when the Pro/Leu genotype was used as the reference. Individuals who carried both putative risk genotypes (Thr/Met and Met/Met for T439M and Pro/Leu for P757L) showed an adjusted OR of 4.95 (95% CI 1.56-15.7) compared with those who carried both low risk genotypes. Analysis of microsatellite instability (MSI) revealed that tumors from individuals who carried both putative risk genotypes tended to have a higher frequency of MSI positives than those from patients who carried both low risk genotypes, although a significant correlation was not found between EXO1 genotype and MSI status. This is the first report to provide evidence for an association of EXO1 gene polymorphisms with colorectal cancer risk. The EXO1 genotypes were not associated with any clinicopathological characteristics in colorectal cancer patients. PMID- 15550455 TI - The signal transduction networks required for phosphorylation of STAT1 at Ser727 in mouse epidermal JB6 cells in the UVB response and inhibitory mechanisms of tea polyphenols. AB - Signal transducers and activators of transcription (STATs) play a critical role in signal transduction pathways. STATs are a family of cytoplasmic proteins with roles as signal messengers and transcription factors that participate in normal cellular responses to cytokines and growth factors. Phosphorylation of STAT1 at Ser727 is essential for its activation and occurs in response to stress signals, inflammation or infection. We observed that UVB induced phosphorylation of STAT1 (Ser727) in mouse epidermal JB6 Cl41 cells. This stimulation was inhibited by PD98059 and UO126, wortmannin, LY294002, SB202190 and SP600125 and dominant negative mutants of ERK2 (DNM-ERK2), p38 (DNM-p38) and JNK1 (DNM-JNK1). The response was absent in Jnk1(-/-) or Jnk2(-/-) knockout cells, but was unaffected by a dominant negative mutant of the phosphatidylinositol-3 kinase (PI-3K) p85 subunit (DNM-Deltap85). STAT1 (Ser727) phosphorylation was also blocked in a Rsk2(-) cell line. In Pdk1(-/-) cells STAT1 was not activated by UVB stimulation compared with strong activation in Pdk1(+/+) cells. Our data indicate that phosphorylation of STAT1 (Ser727) occurs through PI-3K, ERKs, p38 kinase, JNKs, PDK1 and p90RSK2 in the cellular response to UVB. We also show an inhibitory effect of theaflavins and EGCG on UVB-induced STAT1 (Ser727), ERKs, JNKs, PDK1 and p90RSK2 phosphorylation. PMID- 15550456 TI - Green vegetables, red meat and colon cancer: chlorophyll prevents the cytotoxic and hyperproliferative effects of haem in rat colon. AB - Diets high in red meat and low in green vegetables are associated with increased colon cancer risk. This association might be partly due to the haem content of red meat. In rats, dietary haem is metabolized in the gut to a cytotoxic factor that increases colonic cytotoxicity and epithelial proliferation. Green vegetables contain chlorophyll, a magnesium porphyrin structurally analogous to haem. We studied whether green vegetables inhibit the unfavourable colonic effects of haem. First, rats were fed a purified control diet or purified diets supplemented with 0.5 mmol haem/kg, spinach (chlorophyll concentration 1.2 mmol/kg) or haem plus spinach (n = 8/group) for 14 days. In a second experiment we also studied a group that received haem plus purified chlorophyll (1.2 mmol/kg). Cytotoxicity of faecal water was determined with a bioassay and colonic epithelial cell proliferation was quantified in vivo by [methyl-(3)H]thymidine incorporation into newly synthesized DNA. Exfoliation of colonocytes was measured as the amount of rat DNA in faeces. In both studies haem increased cytotoxicity of the colonic contents approximately 8-fold and proliferation of the colonocytes almost 2-fold. Spinach or an equimolar amount of chlorophyll supplement in the haem diet inhibited these haem effects completely. Haem clearly inhibited exfoliation of colonocytes, an effect counteracted by spinach and chlorophyll. Finally, size exclusion chromatography showed that chlorophyll prevented formation of the cytotoxic haem metabolite. We conclude that green vegetables may decrease colon cancer risk because chlorophyll prevents the detrimental, cytotoxic and hyperproliferative colonic effects of dietary haem. PMID- 15550457 TI - Lactate and force production in skeletal muscle. AB - Lactic acid accumulation is generally believed to be involved in muscle fatigue. However, one study reported that in rat soleus muscle (in vitro), with force depressed by high external K(+) concentrations a subsequent incubation with lactic acid restores force and thereby protects against fatigue. However, incubation with 20 mm lactic acid reduces the pH gradient across the sarcolemma, whereas the gradient is increased during muscle activity. Furthermore, unlike active muscle the Na(+)-K(+) pump is not activated. We therefore hypothesized that lactic acid does not protect against fatigue in active muscle. Three incubation solutions were used: 20 mM Na-lactate (which acidifies internal pH), 12 mM Na-lactate +8 mm lactic acid (which mimics the pH changes during muscle activity), and 20 mM lactic acid (which acidifies external pH more than internal pH). All three solutions improved force in K(+)-depressed rat soleus muscle. The pH regulation associated with lactate incubation accelerated the Na(+)-K(+) pump. To study whether the protective effect of lactate/lactic acid is a general mechanism, we stimulated muscles to fatigue with and without pre-incubation. None of the incubation solutions improved force development in repetitively stimulated muscle (Na-lactate had a negative effect). It is concluded that although lactate/lactic acid incubation regains force in K(+)-depressed resting muscle, a similar incubation has no or a negative effect on force development in active muscle. It is suggested that the difference between the two situations is that lactate/lactic acid removes the negative consequences of an unusual large depolarization in the K(+)-treated passive muscle, whereas the depolarization is less pronounced in active muscle. PMID- 15550458 TI - Cellular sources, targets and actions of constitutive nitric oxide in the magnocellular neurosecretory system of the rat. AB - Nitric oxide (NO) is a key activity-dependent modulator of the magnocellular neurosecretory system (MNS) during conditions of high hormonal demand. In addition, recent studies support the presence of a functional constitutive NO tone. The aim of this study was to identify the cellular sources, targets, signalling mechanisms and functional relevance of constitutive NO production within the supraoptic nucleus (SON). Direct visualization of intracellular NO, along with neuronal nitric oxide synthase (nNOS) and cGMP immunohistochemistry, was used to study the cellular sources and targets of NO within the SON, respectively. Our results support the presence of a strong NO basal tone within the SON, and indicate that vasopressin (VP) neurones constitute the major neuronal source and target of basal NO. NO induced-fluorescence and cGMP immunoreactivity (cGMPir) were also found in the glia and microvasculature of the SON, suggesting that they contribute as sources/targets of NO within the SON. cGMPir was also found in association with glutamic acid decarboxylase 67 (GAD67)- and vesicular glutamate transporter 2 (VGLUT2)-positive terminals. Glutamate, acting on NMDA and possibly AMPA receptors, was found to be an important neurotransmitter driving basal NO production within the SON. Finally, electrophysiological recordings obtained from SON neurones in a slice preparation indicated that constitutive NO efficiently restrains ongoing firing activity of these neurones. Furthermore, phasically active (putative VP) and continuously firing neurones appeared to be influenced by NO originating from different sources. The potential roles for basal NO as an autocrine signalling molecule, and one that bridges neuronal-glial-vascular interactions within the MNS are discussed. PMID- 15550459 TI - Increased ratio of rapsyn to ACh receptor stabilizes postsynaptic receptors at the mouse neuromuscular synapse. AB - The metabolic turnover of nicotinic ACh receptors (AChR) at the neuromuscular synapse is regulated over a tenfold range by innervation status, muscle electrical activity and neural agrin, but the downstream effector of such changes has not been defined. The AChR-associated protein rapsyn is essential for forming AChR clusters during development. Here, rapsyn was tagged with enhanced green fluorescent protein (EGFP) to begin to probe its influence at the adult synapse. In C2 myotubes, rapsyn-EGFP participated with AChR in agrin-induced AChR cluster formation. When electroporated into the tibialis anterior muscle of young adult mice, rapsyn-EGFP accumulated in discrete subcellular structures, many of which colocalized with Golgi markers, consistent with the idea that rapsyn assembles with AChR in the exocytic pathway. Rapsyn-EGFP also targeted directly to the postsynaptic membrane where it occupied previously vacant rapsyn binding sites, thereby increasing the rapsyn to AChR ratio. At endplates displaying rapsyn-EGFP, the metabolic turnover of AChR (labelled with rhodamine-alpha-bungarotoxin) was slowed. Thus, the metabolic half-life of receptors at the synapse may be modulated by local changes in the subsynaptic ratio of rapsyn to AChR. PMID- 15550460 TI - Intracellular calcium store filling by an L-type calcium current in the basolateral amygdala at subthreshold membrane potentials. AB - The long-term changes that underlie learning and memory are activated by rises in intracellular Ca2+ that activate a number of signalling pathways and trigger changes in gene transcription. Ca2+ rises due to influx via L-type voltage dependent Ca2+ channels (L-VDCCs) and release from intracellular Ca2+ stores have been consistently implicated in the biochemical cascades that underlie the final changes in memory formation. Here, we show that pyramidal neurones in the basolateral amygdala express an L-VDCC that is active at resting membrane potentials. Subthreshold depolarization of neurones either by current injection or summating synaptic potentials led to a sustained rise in cytosolic Ca2+ that was blocked by the dihydropyridine nicardipine. Activation of metabotropic receptors released Ca2+ from intracellular Ca2+ stores. At hyperpolarized potentials, metabotropic-evoked store release ran down with repeated stimulation. Depolarization of cells to -50 mV, or maintaining them at the resting membrane potential, restored release from intracellular Ca2+ stores, an effect that was blocked by nicardipine. These results show that Ca2+ influx via a low-voltage activated L-type Ca2+ current refills inositol 1,4,5-trisphosphate (IP(3)) sensitive intracellular Ca2+ stores, and maintains Ca2+ release and wave generation by metabotropic receptor activation. PMID- 15550461 TI - Structure of cortical microcircuit theory. AB - Recent experimental and theoretical investigations have made considerable advances in three major areas relating to the structural basis of quantitative cortical microcircuit theory. The first concerns the nature of the cellular units, encompassing the increasingly precise identification and progressively more complete listing of the individual cellular species that constitute the various cortical networks. The second element addresses the problem of heterogeneity, including the demonstration of the importance of cell to cell variability within defined interneuronal populations and the application of the Shannon-Wiener diversity index for the quantitative assessment of the number and relative abundance of interneuronal species. The third component relates to the discovery of basic topological principles underlying the circuit wiring, revealing a surprising order in the architectural design of networks. These new advances deepen our understanding of the computational principles embedded in cortical microcircuits, and they also provide novel opportunities for building realistic models of mammalian cortical microcircuits. PMID- 15550462 TI - Regulation of pyruvate dehydrogenase activity and citric acid cycle intermediates during high cardiac power generation. AB - A high rate of cardiac work increases citric acid cycle (CAC) turnover and flux through pyruvate dehydrogenase (PDH); however, the mechanisms for these effects are poorly understood. We tested the hypotheses that an increase in cardiac energy expenditure: (1) activates PDH and reduces the product/substrate ratios ([NADH]/[NAD(+)] and [acetyl-CoA]/[CoA-SH]); and (2) increases the content of CAC intermediates. Measurements were made in anaesthetized pigs under control conditions and during 15 min of a high cardiac workload induced by dobutamine (Dob). A third group was made hyperglycaemic (14 mm) to stimulate flux through PDH during the high work state (Dob + Glu). Glucose and fatty acid oxidation were measured with (14)C-glucose and (3)H-oleate. Compared with control, the high workload groups had a similar increase in myocardial oxygen consumption ( and cardiac power. Dob increased PDH activity and glucose oxidation above control, but did not reduce the [NADH]/[NAD(+)] and [acetyl-CoA]/[CoA-SH] ratios, and there were no differences between the Dob and Dob + Glu groups. An additional group was treated with Dob + Glu and oxfenicine (Oxf) to inhibit fatty acid oxidation: this increased [CoA-SH] and glucose oxidation compared with Dob; however, there was no further activation of PDH or decrease in the [NADH]/[NAD(+)] ratio. Content of the 4-carbon CAC intermediates succinate, fumarate and malate increased 3-fold with Dob, but there was no change in citrate content, and the Dob + Glu and Dob + Glu + Oxf groups were not different from Dob. In conclusion, compared with normal conditions, at high myocardial energy expenditure (1) the increase in flux through PDH is regulated by activation of the enzyme complex and continues to be partially controlled through inhibition by fatty acid oxidation, and (2) there is expansion of the CAC pool size at the level of 4-carbon intermediates that is largely independent of myocardial fatty acid oxidation. PMID- 15550463 TI - Effect of dissociating cytosolic calcium and metabolic rate on intracellular PO2 kinetics in single frog myocytes. AB - The purpose of this investigation was to utilize 2,3-butanedione monoxime (BDM; an inhibitor of contractile activation) to dissociate cytosolic [Ca(2+)] ([Ca(2+)](c)) from the putative respiratory regulators that arise from muscle contraction-induced ATP utilization in order to determine the relative contribution of [Ca(2+)](c) on intracellular P(O(2)) (P(iO(2))) kinetics during the transition from rest to contractions in single skeletal myocytes isolated from Xenopus laevis lumbrical muscle. Myocytes were subjected to electrically induced isometric tetanic contractions (0.25 Hz; 2-min bouts) while peak tension and either [Ca(2+)](c) (n= 7; ratiometric fluorescence microscopy) or P(iO(2)) (n= 7; phosphorescence microscopy) was measured continuously. Cells were studied under both control and 3 mm BDM conditions in randomized order. Initial (control, 100 +/- 0%; BDM, 72.6 +/- 4.6%), midpoint (control, 86.7 +/- 1.8%; BDM, 61.6 +/- 4.1%) and end (control, 85.0 +/- 2.8%; BDM, 57.5 +/- 5.0%) peak tensions (normalized to initial control values) were significantly reduced (P < 0.05) with BDM compared with control (n= 14). Despite the reduced peak tension, peak [Ca(2+)](c) was not altered (P > 0.05) between control and BDM trials. Thus, the peak tension-to-peak [Ca(2+)](c) ratio was reduced with BDM compared with control. The absolute fall in P(iO(2)) with contractions, which is proportional to the rise in , was significantly reduced with BDM (13.2 +/- 1.3 mmHg) compared with control (22.0 +/- 2.0 mmHg). However, P(iO(2)) onset kinetics (i.e. mean response time (MRT)) was not altered between BDM (66.8 +/- 8.0 s) and control (64.9 +/- 6.3 s) trials. Therefore, the initial rate of change (defined as the fall in P(iO(2))/MRT) was significantly slower in BDM fibres compared with control. These data demonstrate in these isolated single skeletal muscle fibres that unchanged peak [Ca(2+)](c) in the face of reduced metabolic feedback from the contractile sites evoked with BDM did not alter P(iO(2)) onset kinetics in isolated single frog myocytes, suggesting that metabolic signals arising from the contractile sites play a more substantial role than [Ca(2+)](c) in the signalling pathway to oxidative phosphorylation during the transition from rest to repeated tetanic contractions. PMID- 15550464 TI - Neutrophils contribute to muscle injury and impair its resolution after lengthening contractions in mice. AB - We tested the hypotheses that: (1) neutrophil accumulation after contraction induced muscle injury is dependent on the beta(2) integrin CD18, (2) neutrophils contribute to muscle injury and oxidative damage after contraction-induced muscle injury, and (3) neutrophils aid the resolution of contraction-induced muscle injury. These hypotheses were tested by exposing extensor digitorum longus (EDL) muscles of mice deficient in CD18 (CD18(-/-); Itgb2(tm1Bay)) and of wild type mice (C57BL/6) to in situ lengthening contractions and by quantifying markers of muscle inflammation, injury, oxidative damage and regeneration/repair. Neutrophil concentrations were significantly elevated in wild type mice at 6 h and 3 days post-lengthening contractions; however, neutrophils remained at control levels at these time points in CD18-/- mice. These data indicate that CD18 is required for neutrophil accumulation after contraction-induced muscle injury. Histological and functional (isometric force deficit) signs of muscle injury and total carbonyl content, a marker of oxidative damage, were significantly higher in wild type relative to CD18-/- mice 3 days after lengthening contractions. These data show that neutrophils exacerbate contraction-induced muscle injury. After statistically controlling for differences in the force deficit at 3 days, wild type mice also demonstrated a higher force deficit at 7 days, a lower percentage of myofibres expressing embryonic myosin heavy chain at 3 and 7 days, and a smaller cross sectional area of central nucleated myofibres at 14 days relative to CD18-/- mice. These observations suggest that neutrophils impair the restoration of muscle structure and function after injury. In conclusion, neutrophil accumulation after contraction-induced muscle injury is dependent on CD18. Furthermore, neutrophils appear to contribute to muscle injury and impair some of the events associated with the resolution of contraction-induced muscle injury. PMID- 15550465 TI - Alterations in insulin receptor signalling in the rat epitrochlearis muscle upon cessation of voluntary exercise. AB - The major purpose of this study was to elucidate mechanisms by which decreasing enhanced physical activity induces decreased insulin sensitivity in skeletal muscle. Rats with access to voluntary running wheels for 3 weeks had their wheels locked for 5 h (WL5), 29 h (WL29), or 53 h (WL53); a separate group of rats never had wheel access (sedentary, SED). Relative to WL5, submaximal insulin-stimulated 2-deoxyglucose uptake into the epitrochlearis muscle was lower in WL53 and SED. Insulin binding, insulin receptor beta-subunit (IRbeta) protein level, submaximal insulin-stimulated IRbeta tyrosine phosphorylation, and glucose transporter-4 protein level were each lower in both WL53 and SED than in WL5 and WL29. Akt/protein kinase B Ser(473) phosphorylation was lower in WL53 and SED than in WL5. Protein levels of protein tyrosine phosphatase-1B, Src homology phosphatase 2, and protein kinase C- did not vary among groups. The amount of protein tyrosine phosphatase-1B, Src homology phosphatase-2, and protein kinase C- associated with IRbeta in insulin-stimulated muscle also did not differ among the four groups. The mean of SED and WL53 had a significantly higher IRbeta associated protein tyrosine phosphatase-1B than the mean of WL5 and WL29. The enclosure of multiple changes (decreases in insulin binding, IRbeta protein, IRbeta tyrosine phosphorylation, and glucose transporter-4 protein) in the epitrochlearis muscle within the 29th to 53rd hour after cessation of voluntary wheel running raises the possibility that a single regulatory event could be responsible for the coordinated decrease. PMID- 15550466 TI - Directional analysis of coherent oscillatory field potentials in the cerebral cortex and basal ganglia of the rat. AB - Population activity in cortico-basal ganglia circuits is synchronized at different frequencies according to brain state. However, the structures that are likely to drive the synchronization of activity in these circuits remain unclear. Furthermore, it is not known whether the direction of transmission of activity is fixed or dependent on brain state. We have used the directed transfer function (DTF) to investigate the direction in which coherent activity is effectively driven in cortico-basal ganglia circuits. Local field potentials (LFPs) were simultaneously recorded in the subthalamic nucleus (STN), globus pallidus (GP) and substantia nigra pars reticulata (SNr), together with the ipsilateral frontal electrocorticogram (ECoG) of anaesthetized rats. Directional analysis was performed on recordings made during robust cortical slow-wave activity (SWA) and "global activation". During SWA, there was coherence at approximately 1 Hz between ECoG and basal ganglia LFPs, with much of the coherent activity directed from cortex to basal ganglia. There were similar coherent activities at approximately 1 Hz within the basal ganglia, with more activity directed from SNr to GP and STN, and from STN to GP rather than vice versa. During global activation, peaks in coherent activity were seen at higher frequencies (15-60 Hz), with most coherence also directed from cortex to basal ganglia. Within the basal ganglia, however, coherence was predominantly directed from GP to STN and SNr. Together, these results highlight a lead role for the cortex in activity relationships with the basal ganglia, and further suggest that the effective direction of coupling between basal ganglia nuclei is dynamically organized according to brain state, with activity relationships involving the GP displaying the greatest capacity to change. PMID- 15550467 TI - Functional diversity of electrogenic Na+-HCO3- cotransport in ventricular myocytes from rat, rabbit and guinea pig. AB - The Na(+)-HCO(3)(-) cotransporter (NBC) is an important sarcolemmal acid extruder in cardiac muscle. The characteristics of NBC expressed functionally in heart are controversial, with reports suggesting electroneutral (NBCn; 1HCO(3)(-) : 1Na(+); coupling coefficient N= 1) or electrogenic forms of the transporter (NBCe; equivalent to 2HCO(3)(-) : 1Na(+); N= 2). We have used voltage-clamp and epifluorescence techniques to compare NBC activity in isolated ventricular myocytes from rabbit, rat and guinea pig. Depolarization (by voltage clamp or hyperkalaemia) reversibly increased steady-state pH(i) while hyperpolarization decreased it, effects seen only in CO(2)/HCO(3)(-)-buffered solutions, and blocked by S0859 (cardiac NBC inhibitor). Species differences in amplitude of these pH(i) changes were rat > guinea pig approximately rabbit. Tonic depolarization (-140 mV to -0 mV) accelerated NBC-mediated pH(i) recovery from an intracellular acid load. At 0 mV, NBC-mediated outward current at resting pH(i) was +0.52 +/- 0.05 pA pF(-1) (rat, n= 5), +0.26 +/- 0.05 pA pF(-1) (guinea pig, n= 5) and +0.10 +/- 0.03 pA pF(-1) (rabbit, n= 9), with reversal potentials near 100 mV, consistent with N= 2. The above results indicate a functionally active voltage-sensitive NBCe in these species. Voltage-clamp hyperpolarization negative to the reversal potential for NBCe failed, however, to terminate or reverse NBC mediated pH(i)-recovery from an acid load although it was slowed significantly, suggesting electroneutral NBC may also be operational. NBC-mediated pH(i) recovery was associated with a rise of [Na(+)](i) at a rate approximately 25% of that mediated via NHE, and consistent with an apparent NBC stoichiometry between N= 1 and N= 2. In conclusion, NBCe in the ventricular myocyte displays considerable functional variation among the three species tested (greatest in rat, least in rabbit) and may coexist with some NBCn activity. PMID- 15550468 TI - Dissociation of slow waves and fast oscillations above 200 Hz during GABA application in rat somatosensory cortex. AB - Fast electrical oscillations (FOs; > 200 Hz), superimposed on vibrissa-evoked slow potentials, may support rapid sensory integration in neocortex. Yet, while it is well established that the positive/negative (P1/N1) slow wave of the somatosensory evoked potential primarily reflects sequential activation of supragranular and infragranular pyramidal cells mediated chiefly via excitatory chemical synaptic pathways, little is known about the generation of FOs. In this study, laminar current source-density analysis and principal component analysis indicated that FOs are generated by two dipolar current sources situated in the supra- and infragranular layers, similar in laminar location to the two current dipoles associated with the slow wave. However, exogenous GABA application reversibly abolished the N1 slow wave, leaving the P1 intact, while the FO was unaffected by GABA. Furthermore, reductions in both supra- and infragranular cortical unit discharge during application of GABA suggests that FO generation is not dependent on the same intracortical synaptic circuits that are associated with the N1 slow wave. These data suggest a marked functional dissociation between neural mechanisms underlying the slow and fast components of the vibrissa evoked response. PMID- 15550469 TI - Attenuation of conducted vasodilatation in rat mesenteric arteries during hypertension: role of inwardly rectifying potassium channels. AB - The present study was designed to elucidate whether the conduction of vasomotor responses mediated by endothelium-derived hyperpolarizing factor (EDHF) in rat mesenteric arteries is altered during hypertension. Iontophoresed acetylcholine (ACh; 500 ms) caused EDHF-mediated hyperpolarization and vasodilatation at the local site and these responses spread through the endothelium to remote sites in 12-week-old Wistar-Kyoto rats (WKY). Conducted responses were significantly attenuated in age-matched spontaneously hypertensive rats (SHR) although the rate of decay with distance did not change. Inhibition of inwardly rectifying potassium (Kir) channels (30 microM barium) eliminated the difference between WKY and SHR by attenuating conducted responses in WKY but not SHR. At the local site, barium (30 microM) significantly reduced the duration but not the amplitude of ACh-induced hyperpolarization in WKY only. Barium had no effect when the iontophoretic stimulus was reduced to 350 ms. After blockade of EDHF in SHR, ACh elicited a depolarization which our indirect data suggest spreads along the vessel in the endothelium. Messenger RNA expression of Kir2.0 genes did not differ between the strains nor did the amplitude of K(+)-induced hyperpolarization, which was abolished by disruption of the endothelium. Immunohistochemistry revealed a decrease in connexin (Cx)37 but not Cx40 or Cx43 protein in endothelial cells of SHR compared to WKY. Results suggest that conduction of EDHF-mediated responses in WKY, but not in SHR, is facilitated by activation of Kir channels at the site of ACh application and not by differences in endothelial connexin expression. Lack of Kir channel involvement in hypertension may result from reduction in the duration of the hyperpolarization due to the development of ACh-mediated depolarization, rather than to any difference in Kir subunit expression or function. PMID- 15550470 TI - Increased insulin translation from an insulin splice-variant overexpressed in diabetes, obesity, and insulin resistance. AB - Type 2 diabetes occurs when pancreatic beta-cells become unable to compensate for the underlying insulin resistance. Insulin secretion requires beta-cell insulin stores to be replenished by insulin biosynthesis, which is mainly regulated at the translational level. Such translational regulation often involves the 5' untranslated region. Recently, we identified a human insulin splice-variant (SPV) altering only the 5'-untranslated region and conferring increased translation efficiency. We now describe a mouse SPV (mSPV) that is found in the cytoplasm and exhibits increased translation efficiency resulting in more normal (prepro)insulin protein per RNA. The RNA stability of mSPV is not increased, but the predicted secondary RNA structure is altered, which may facilitate translation. To determine the role of mSPV in insulin resistance and diabetes, mSPV expression was measured by quantitative real-time RT-PCR in islets from three diabetic and/or insulin-resistant, obese and nonobese, mouse models (BTBRob/ob, C57BL/6ob/ob, and C57BL/6azip). Interestingly, mSPV expression was significantly higher in all diabetic/insulin-resistant mice compared with wild type littermates and was dramatically induced in primary mouse islets incubated at high glucose. This raises the possibility that the mSPV may represent a compensatory beta-cell mechanism to enhance insulin biosynthesis when insulin requirements are elevated by hyperglycemia/insulin resistance. PMID- 15550471 TI - Overexpression of an N-terminally truncated isoform of the nuclear receptor coactivator amplified in breast cancer 1 leads to altered proliferation of mammary epithelial cells in transgenic mice. AB - Amplified in breast cancer 1 (AIB1, also known as ACTR, SRC-3, RAC-3, TRAM-1, p/CIP) is a member of the p160 nuclear receptor coactivator family involved in transcriptional regulation of genes activated through steroid receptors, such as estrogen receptor alpha (ER(alpha)). The AIB1 gene and a more active N-terminally deleted isoform (AIB1-Delta3) are overexpressed in breast cancer. To determine the role of AIB1-Delta3 in breast cancer pathogenesis, we generated transgenic mice with human cytomegalovirus immediate early gene 1 (hCMVIE1) promoter-driven over-expression of human AIB1/ACTR-Delta3 (CMVAIB1/ACTR-Delta3 mice). AIB1/ACTR Delta3 transgene mRNA expression was confirmed in CMV-AIB1/ACTR-Delta3 mammary glands by in situ hybridization. These mice demonstrated significantly increased mammary epithelial cell proliferation (P < 0.003), cyclin D1 expression (P = 0.002), IGF-I receptor protein expression (P = 0.026), mammary gland mass (P < 0.05), and altered expression of CCAAT/enhancer binding protein isoforms (P = 0.029). At 13 months of age, mammary ductal ectasia was found in CMV-AIB1/ACTR Delta3 mice, but secondary and tertiary branching patterns were normal. There were no changes in the expression patterns of either ER(alpha) or Stat5a, a downstream mediator of prolactin signaling. Serum IGF-I levels were not altered in the transgenic mice. These data indicate that overexpression of the AIB1/ACTR Delta3 isoform resulted in altered mammary epithelial cell growth. The observed changes in cell proliferation and gene expression are consistent with alterations in growth factor signaling that are thought to contribute to either initiation or progression of breast cancer. These results are consistent with the hypothesis that the N-terminally deleted isoform of AIB1 can play a role in breast cancer development and/or progression. PMID- 15550472 TI - Metrologic traceability of total thyroxine measurements in human serum: efforts to establish a network of reference measurement laboratories. AB - BACKGROUND: Assuring/demonstrating metrologic traceability of in vitro diagnostics necessitates the availability of measurand-specific reference measurement systems (RMSs) and the possibility for industry to work with competent reference measurement laboratories (RMLs). Here we report the results of a European project to investigate the feasibility of developing a RMS for serum total thyroxine. METHODS: Four candidate RMLs (cRMLs) developed/implemented variants of a candidate reference measurement procedure (cRMP) based on isotope dilution-liquid chromatography-mass spectrometry. The sole constraint implemented was calibration with a common thyroxine primary calibrator. The RMPs were externally validated and assessed for comparability in round-robin trials using common samples, i.e., 5 lyophilized and 33 frozen native sera. At the same time, the performance of the cRMLs organized in a network was assessed. For uniform external quality assessment, common performance specifications were agreed on. RESULTS: All cRMLs performed the cRMPs with fulfillment of the predefined specifications: total and between-laboratory CVs < or =2.0% and 2.5%, respectively, and a systematic deviation < or =0.9%, estimated with a target assigned from the mean of means obtained by the cRMLs. The mean expanded uncertainty for value assignment to the native sera was 2.1%. CONCLUSIONS: A network of cRMLs, with externally conformed competence to properly perform RMPs, has been established. Performance specifications were defined and will form the basis for admittance of new network members. A serum panel, successfully targeted during the validation process, is available for split-sample measurements with commercial routine measurement procedures. The model can now be used for other measurands for which traceability to the Systeme International d'Unites is needed. PMID- 15550473 TI - Increased plasma lipid-poor apolipoprotein A-I in patients with coronary artery disease. AB - BACKGROUND: Pre-beta 1-HDL participates in a cyclic process involved in the retrieval of cholesterol from peripheral tissues. Although pre-beta 1-HDL can be measured by two-dimensional electrophoresis or crossed immunoelectrophoresis, these methods are time-consuming and require technical expertise. In this study, we separated plasma lipid-poor apolipoprotein A-I (apo A-I) by high-performance size-exclusion chromatography. METHODS: We measured plasma lipid-poor apo A-I in 20 male patients with coronary artery disease [CAD; mean (SD) age, 64.0 (18) years] and 15 male controls [54.7 (17) years] and in 7 female CAD patients [70.3 (7.7) years] and 9 female controls [65.1 (4.7) years]. RESULTS: Lipid-poor apo A I was most stable when stored at -80 degrees C in the presence of aprotinin (final concentration, 50 kIU/L). The lipid-poor apo A-I concentration decreased during incubation at 37 degrees C, and this was not prevented by the addition of 2 mmol/L of the lecithin:cholesterol acyltransferase (LCAT) inhibitor 5,5' dithiobis(2-nitrobenzoic acid). Lipid-poor apo A-I was significantly higher in CAD patients than in controls [38.3 (7.9) mg/L for male CAD patients vs 29.3 (7.3) mg/L for male controls; 43.3 (11) mg/L for female CAD patients vs 27.1 (7.4) mg/L for female controls (P <0.01 for both)]. There were no significant differences in LCAT activity or cholesteryl ester transfer protein (CETP) concentration between patients and controls. Moreover, the plasma lipid-poor apo A-I concentration was not significantly correlated with LCAT or CETP activities. CONCLUSIONS: Although the production of lipid-poor apo A-I in plasma is not fully understood, our results indicate that lipid-poor apo A-I could be used as a marker for arteriosclerosis and demonstrate that it is not identical to the pre beta1-HDL measured by other methods. PMID- 15550474 TI - Prognostic value of an early soluble L-selectin (sCD62L) assay for risk assessment in blunt multiple trauma: a metaanalysis. AB - BACKGROUND: After severe trauma, decreased plasma concentrations of the soluble adhesion molecule L-selectin (sCD62L) have been linked to an increased incidence of lung failure and multiorgan dysfunction syndrome (MODS). Individual studies have had conflicting results, however. We examined multiple studies in an attempt to determine whether early sCD62L concentrations are predictive of major complications after severe trauma. METHODS: We performed a systematic review of six electronic databases and a manual search for clinical studies comparing outcomes of multiply injured patients (Injury Severity Score > or =16) depending on their early sCD62L blood concentrations. Because of various outcome definitions, acute lung injury (ALI) and adult respiratory distress syndrome (ARDS) were studied as a composite endpoint. Weighted mean differences (WMDs) in sCD62L concentrations were calculated between individuals with and without complications by fixed- and random-effects models. RESULTS: Altogether, 3370 citations were identified. Seven prospective studies including 350 patients were eligible for data synthesis. Published data showed the discriminatory features of sCD62L but did not allow for calculation of measures of test accuracy. Three of four studies showed lower early sCD62L concentrations among individuals progressing to ALI and ARDS (WMD = -229 microg/L; 95% confidence interval, -476 to 18 microg/L). No differences in sCD62L concentrations were noted among patients with or without later MODS. Nonsurvivors had significantly lower early sCD62L plasma concentrations (WMD = 121 microg/L; 95% confidence interval, 63-179 microg/L), but little information was available on potential confounders in this group. CONCLUSIONS: Early decreased soluble L-selectin concentrations after multiple trauma may signal an increased likelihood of lung injury and ARDS. The findings of this metaanalysis warrant a large cohort study to develop selectin based models targeting the risk of inflammatory complications. PMID- 15550475 TI - Serum S-100B as an indicator of early postoperative deterioration after meningioma surgery. AB - BACKGROUND: S-100B protein is an established serum marker of primary and secondary brain damage in head injury and stroke. Despite major progress in neurophysiologic monitoring, there are still difficulties in the early identification and quantification of evolving edema or trauma after craniotomy for tumor. In this study we aimed to correlate serum S-100B values with early postoperative neurologic course as well as late outcome in meningioma surgery. METHODS: We enrolled 50 consecutive patients who underwent meningioma resection. Serum S-100B was measured preoperatively and postcraniotomy for 7 consecutive days. Twenty-five patients (50%) developed immediate postoperative neurologic deterioration, and 15 (30%) had unfavorable 6-month outcomes. We used the Mann Whitney U-test to assess the association of S-100B with all variables of interest. We used multiple logistic regression to search for the most significant predictor of postoperative deterioration. RESULTS: Increased S-100B was highly correlated with larger tumors, intraoperative difficulties, postcraniotomy acute deterioration, and long-term poor outcome. In addition, multiple logistic regression showed that age, sex, site, preoperative edema, history of meningioma resection, extent of resection, and histologic type did not correlate with postoperative increases in S-100B. Furthermore, patients with postoperative S 100B values >0.4 microg/L had increased risk of deterioration (relative risk = 9.0; 95% confidence interval, 2.4-34; P <0.0001) and of poor ultimate outcome (relative risk = 11; 95% confidence interval, 1.6-77; P = 0.002). CONCLUSIONS: After meningioma excision, postcraniotomy increases in serum S-100B appear to be an early indicator of short-term postoperative neurologic deterioration and of a poor longer-term outcome. PMID- 15550476 TI - Enhancement of sensitivity and resolution of surface-enhanced laser desorption/ionization time-of-flight mass spectrometric records for serum peptides using time-series analysis techniques. AB - BACKGROUND: Measurement of peptide/protein concentrations in biological samples for biomarker discovery commonly uses high-sensitivity mass spectrometers with a surface-processing procedure to concentrate the important peptides. These time-of flight (TOF) instruments typically have low mass resolution and considerable electronic noise associated with their detectors. The net result is unnecessary overlapping of peaks, apparent mass jitter, and difficulty in distinguishing mass peaks from background noise. Many of these effects can be reduced by processing the signal using standard time-series background subtraction, calibration, and filtering techniques. METHODS: Surface-enhanced laser desorption/ionization (SELDI) spectra were acquired on a PBS II instrument from blank, hydrophobic, and IMAC-Cu ProteinChip arrays (Ciphergen Biosystems, Inc.) incubated with calibration peptide mixtures or pooled serum. TOF data were recorded after single and multiple laser shots at different positions. Correlative analysis was used for time-series calibration. Target filters were used to suppress noise and enhance resolution after baseline removal and noise rescaling. RESULTS: The developed algorithms compensated for the electronic noise attributable to detector overload, removed the baseline caused by charge accumulation, detected and corrected mass peak jitter, enhanced signal amplitude at higher masses, and improved the resolution by using a deconvolution filter. CONCLUSIONS: These time series techniques, when applied to SELDI-TOF data before any peak identification procedure, can improve the data to make the peak identification process simpler and more robust. These improvements may be applicable to most TOF instrumentation that uses analog (rather than counting) detectors. PMID- 15550477 TI - N-terminal pro-B-type natriuretic peptide as an indicator of possible cardiovascular disease in severely obese individuals: comparison with patients in different stages of heart failure. AB - BACKGROUND: Mild stages of heart failure might be difficult to diagnose in severely obese individuals with a body mass index (BMI) >40 kg/m(2). Measurement of the N-terminal fragment of pro-B-type natriuretic peptide (NT-proBNP) is feasible for detecting cardiac impairment. The aims of our study were to measure NT-proBNP in plasma of severely obese patients and to compare the results with results for patients in different stages of manifest cardiac dysfunction. METHODS: In 61 severely obese individuals (median BMI, 43.2 kg/m(2)) and 96 nonobese patients with existing heart failure [classified into New York Heart Association (NYHA) classes I-IV], NT-proBNP was measured in the fasting condition. A medical history, physical examination, electrocardiography, blood chemistry, and chest x-ray were performed in the obese group. In addition, echocardiography was performed in the NYHA group. RESULTS: In obese individuals, NT-proBNP was increased to a median of 356 (interquartile range, 221-458) pmol/L [854 (530-1099) ng/L] and was comparable (P >0.05) to the median value for NYHA I patients {289 (258-451) pmol/L [694 (619-1082) ng/L]}, but was significantly lower than in the other NYHA groups (P <0.001 for each). CONCLUSION: The prognostic relevance of increased NT-proBNP for risk of developing cardiac insufficiency in severely obese patients needs to be further evaluated. PMID- 15550478 TI - Optimized spectrophotometric assay for the completely activated pyruvate dehydrogenase complex in fibroblasts. AB - BACKGROUND: Analysis of the pyruvate dehydrogenase complex (PDHc) activity in human skin fibroblasts is hampered by low enzyme activity in the cells. The most commonly used radiochemical method detects the formation of (14)CO(2), an endproduct of the E1 component of PDHc, from [1-(14)C]pyruvate. METHODS: We report a spectrophotometric method for the analysis of PDHc activity in fibroblasts based on detection of NADH formation via a p-iodonitrotetrazolium violet (INT)-coupled system. We investigated in detail the specific requirements of this assay, such as cofactor requirements and the effects of suggested stimulatory compounds and different cell disruption procedures. The reliability of the optimized assay was studied by investigation of patients previously diagnosed with PDHc deficiency and by comparison with results from the radiochemical method. RESULTS: Mean (SD) total PDHc activities were 136 (31) and 58 (21) mU/U of citrate synthase in fibroblast homogenates from 10 healthy volunteers and 7 PDHc-deficient patients, respectively, by the spectrophotometric assay. Similar results were obtained in a mitochondrial fraction. Dithiothreitol (DTT) increased the nonspecific inhibitor-insensitive rate with less pronounced effect on the specific rate of PDHc activity. Administration of DTT increased PDHc activity to 193 (3)% of control activity (without DTT), but decreased the inhibitor-sensitive rate from 99 (0.3)% (without DTT) to 69 (2)% (with 0.3 mmol/L DTT). CONCLUSION: The simple, optimized spectrophotometric assay for PDHc analysis allows reliable investigation of the enzyme complex in human skin fibroblasts. PMID- 15550479 TI - Proteomic fingerprints for potential application to early diagnosis of severe acute respiratory syndrome. AB - BACKGROUND: Definitive early-stage diagnosis of severe acute respiratory syndrome (SARS) is important despite the number of laboratory tests that have been developed to complement clinical features and epidemiologic data in case definition. Pathologic changes in response to viral infection might be reflected in proteomic patterns in sera of SARS patients. METHODS: We developed a mass spectrometric decision tree classification algorithm using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. Serum samples were grouped into acute SARS (n = 74; <7 days after onset of fever) and non-SARS [n = 1067; fever and influenza A (n = 203), pneumonia (n = 176); lung cancer (n = 29); and healthy controls (n = 659)] cohorts. Diluted samples were applied to WCX-2 ProteinChip arrays (Ciphergen), and the bound proteins were assessed on a ProteinChip Reader (Model PBS II). Bioinformatic calculations were performed with Biomarker Wizard software 3.1.1 (Ciphergen). RESULTS: The discriminatory classifier with a panel of four biomarkers determined in the training set could precisely detect 36 of 37 (sensitivity, 97.3%) acute SARS and 987 of 993 (specificity, 99.4%) non-SARS samples. More importantly, this classifier accurately distinguished acute SARS from fever and influenza with 100% specificity (187 of 187). CONCLUSIONS: This method is suitable for preliminary assessment of SARS and could potentially serve as a useful tool for early diagnosis. PMID- 15550480 TI - Identifying subtle interrelated changes in functional gene categories using continuous measures of gene expression. AB - MOTIVATION: Analysis of large-scale expression data is greatly facilitated by the availability of gene ontologies (GOs). Many current methods test whether sets of transcripts annotated with specific ontology terms contain an excess of 'changed' transcripts. This approach suffers from two main limitations. First, since gene expression is continuous rather than discrete, designating a gene as changed or unchanged is arbitrary and oblivious to the actual magnitude of the change. Second, by considering only the number of changed genes, finer changes in expression patterns associated with the category may be ignored. Since genes generally participate in multiple networks, widespread and subtle modifications in expression patterns are at least as important as extreme increases/decreases of a few genes. RESULTS: Numerical simulations confirm that incorporating continuous measures of gene expression for all measured transcripts yields detection of considerably more subtle changes. Applying continuous measures to microarray data from brains of mice injected with the Parkinsonian neurotoxin, MPTP, enables detection of changes in various biologically relevant GO terms, many of which are overlooked by discrete approaches. PMID- 15550481 TI - CRAVE: a database, middleware and visualization system for phenotype ontologies. AB - MOTIVATION: A major challenge in modern biology is to link genome sequence information to organismal function. In many organisms this is being done by characterizing phenotypes resulting from mutations. Efficiently expressing phenotypic information requires combinatorial use of ontologies. However tools are not currently available to visualize combinations of ontologies. Here we describe CRAVE (Concept Relation Assay Value Explorer), a package allowing storage, active updating and visualization of multiple ontologies. RESULTS: CRAVE is a web-accessible JAVA application that accesses an underlying MySQL database of ontologies via a JAVA persistent middleware layer (Chameleon). This maps the database tables into discrete JAVA classes and creates memory resident, interlinked objects corresponding to the ontology data. These JAVA objects are accessed via calls through the middleware's application programming interface. CRAVE allows simultaneous display and linking of multiple ontologies and searching using Boolean and advanced searches. PMID- 15550482 TI - Functional characterization of telomerase RNA variants found in patients with hematologic disorders. AB - Human telomerase uses a specific cellular RNA, called hTERC, as the template to synthesize telomere repeats at chromosome ends. Approximately 10% to 15% of patients with aplastic anemia or other bone marrow failure syndromes are carriers of hTERC sequence variants whose functional significance, in most cases, is unknown. We screened 10 reported and 2 newly discovered hTERC variants from such patients and found that 10 of these negatively affected telomerase enzymatic function when they were used to reconstitute telomerase enzymatic function in human cells. Most functional deficits were due to perturbations of hTERC secondary structure and correlated well with the degrees of telomere shortening and reduced telomerase activity observed in peripheral blood lymphocytes of the representative patients. We also found no evidence of dominant-negative activity in any of the mutants. Therefore, loss of telomerase activity and of telomere maintenance resulting from inherited hTERC mutations may limit marrow stem cell renewal and predispose some patients to bone marrow failure. PMID- 15550483 TI - Regulation of tissue factor-induced signaling by endogenous and recombinant tissue factor pathway inhibitor 1. AB - Tissue factor (TF) triggers upstream coagulation signaling via the activation of protease-activated receptors (PARs) of relevance for inflammation and angiogenesis. TF pathway inhibitor 1 (TFPI-1) is the physiologic inhibitor of TF initiated coagulation, but its role in regulating TF signaling is poorly understood. Here, we demonstrate that endogenous, endothelial cell-expressed TFPI 1 controls TF-mediated signaling through PARs. In endothelial cells transduced with TF to mimic exacerbated TF expression in vascular cells, TF-VIIa-Xa ternary complex-dependent activation of PAR1 remained intact when TF-mediated Xa generation was blocked with 2.5 to 5 nM recombinant TFPI-1 (rTFPI-1). Concordantly, inhibition of signaling in PAR1-expressing Chinese hamster ovary (CHO) cells required about 30-fold higher rTFPI-1 concentrations than necessary for anticoagulation. Studies with proteoglycan-deficient CHO cells document a crucial role of accessory receptors in supporting the anticoagulant and antisignaling activities of rTFPI-1. Coexpression of PAR2 with TF enhanced rTFPI mediated inhibition of TF-VIIa-Xa-mediated PAR1 signaling, suggesting an unexpected role of PAR2 in the inhibitory control of TF signaling. These experiments are of potential significance for the limited therapeutic benefit of rTFPI-1 in systemic inflammation and recommend caution in using anticoagulant potency as a measure to predict how efficacious TF-directed inhibitors block cell signaling during initiation of coagulation. PMID- 15550484 TI - Rituximab plus infusional cyclophosphamide, doxorubicin, and etoposide in HIV associated non-Hodgkin lymphoma: pooled results from 3 phase 2 trials. AB - Evidence suggests that infusional therapy is a more effective means for administering cytotoxic therapy than intravenous bolus therapy for lymphoma and offers greater potential for therapeutic synergy with rituximab, which has a long half-life. We pooled the results of 3 prospective phase 2 trials evaluating rituximab in combination with 96-hour infusion of cyclophosphamide (187.5-200 mg/m2 per day), doxorubicin (12.5 mg/m2 per day), and etoposide (60 mg/m2 per day) (R-CDE) plus granulocyte-colony-stimulating factor (G-CSF) in 74 patients with HIV-associated, B-cell non-Hodgkin lymphoma, of whom 56 (76%) patients received concurrent highly active antiretroviral therapy (HAART). The complete remission (CR) rate was 70% (95% confidence interval [CI], 59%-81%), and the estimated 2-year failure-free survival and overall survival rates were 59% (95% CI, 47%-71%) and 64% (95% CI, 52%-76%), respectively. Ten (14%) patients had opportunistic infections during or within 3 months of the end of R-CDE, and 17 (23%) patients developed nonopportunistic infections after that time. Six (8%) patients died because of infection; 2 (3%) of those infections were bacterial sepsis during R-CDE, and 4 (5%) were opportunistic infections that occurred between 2 and 8 months after the completion of R-CDE. R-CDE produced a 70% CR rate and a 59% 2-year failure-free survival rate in patients with HIV-associated lymphoma. Consistent with other reports, adding rituximab to cytotoxic therapy in this population may increase the risk for life-threatening infection. Further studies evaluating rituximab in combination with infusional chemotherapy are warranted, but caution is advised. PMID- 15550485 TI - Vascular endothelial growth factor promotes sensitivity to ultraviolet B-induced cutaneous photodamage. AB - Acute ultraviolet B (UVB) irradiation of the skin results in erythema, vasodilation, edema, and angiogenesis, which is associated with the expression of vascular endothelial growth factor (VEGF) by epidermal keratinocytes. It is unclear, however, whether VEGF is required for the damage or repair process that occurs in the skin on UVB exposure. We subjected transgenic mice that overexpress VEGF, and their wild-type littermates, to graded doses of acute UVB irradiation. The skin of VEGF-overexpressing mice was highly photosensitive and became erythematic when exposed to half the UVB dose required to induce erythema in wild type mice. Erythema was associated with proliferating dermal endothelial cells, cutaneous edema, and inflammatory cell infiltration. When subjected to 10 weeks of low-level UVB irradiation, no major changes were observed in wild-type mice, whereas VEGF transgenic mice developed skin damage associated with degradation of the dermal matrix and enhanced vascularization. Systemic treatment with an anti VEGF blocking antibody reduced the sensitivity of wild-type mice to acute UVB irradiation without inhibiting post-UVB repair. Our results reveal that VEGF promotes the cutaneous damage that occurs after UVB exposure and that the VEGF signaling pathway might serve as a novel target for the prevention of UVB-induced photodamage. PMID- 15550486 TI - Jagged1-dependent Notch signaling is dispensable for hematopoietic stem cell self renewal and differentiation. AB - Jagged1-mediated Notch signaling has been suggested to be critically involved in hematopoietic stem cell (HSC) self-renewal. Unexpectedly, we report here that inducible Cre-loxP-mediated inactivation of the Jagged1 gene in bone marrow progenitors and/or bone marrow (BM) stromal cells does not impair HSC self renewal or differentiation in all blood lineages. Mice with simultaneous inactivation of Jagged1 and Notch1 in the BM compartment survived normally following a 5FU-based in vivo challenge. In addition, Notch1-deficient HSCs were able to reconstitute mice with inactivated Jagged1 in the BM stroma even under competitive conditions. In contrast to earlier reports, these data exclude an essential role for Jagged1-mediated Notch signaling during hematopoiesis. PMID- 15550487 TI - Factor IX variants improve gene therapy efficacy for hemophilia B. AB - Intramuscular injection of adeno-associated viral (AAV) vector to skeletal muscle of humans with hemophilia B is safe, but higher doses are required to achieve therapeutic factor IX (F.IX) levels. The efficacy of this approach is hampered by the retention of F.IX in muscle extracellular spaces and by the limiting capacity of muscle to synthesize fully active F.IX at high expression rates. To overcome these limitations, we constructed AAV vectors encoding F.IX variants for muscle- or liver-directed expression in hemophilia B mice. Circulating F.IX levels following intramuscular injection of AAV-F.IX-K5A/V10K, a variant with low affinity to extracellular matrix, were 2-5 fold higher compared with wild-type (WT) F.IX, while the protein-specific activities remained similar. Expression of F.IX-R338A generated a protein with 2- or 6-fold higher specific activity than F.IX-WT following vector delivery to skeletal muscle or liver, respectively. F.IX WT and variant forms provide effective hemostasis in vivo upon challenge by tail clipping assay. Importantly, intramuscular injection of AAV-F.IX variants did not trigger antibody formation to F.IX in mice tolerant to F.IX-WT. These studies demonstrate that F.IX variants provide a promising strategy to improve the efficacy for a variety of gene-based therapies for hemophilia B. PMID- 15550488 TI - Antileukemic activity of rapamycin in acute myeloid leukemia. AB - The mammalian target of rapamycin (mTOR) is a key regulator of growth and survival in many cell types. Its constitutive activation has been involved in the pathogenesis of various cancers. In this study, we show that mTOR inhibition by rapamycin strongly inhibits the growth of the most immature acute myeloid leukemia (AML) cell lines through blockade in G0/G1 phase of the cell cycle. Accordingly, 2 downstream effectors of mTOR, 4E-BP1 and p70S6K, are phosphorylated in a rapamycin-sensitive manner in a series of 23 AML cases. Interestingly, the mTOR inhibitor markedly impairs the clonogenic properties of fresh AML cells while sparing normal hematopoietic progenitors. Moreover, rapamycin induces significant clinical responses in 4 of 9 patients with either refractory/relapsed de novo AML or secondary AML. Overall, our data strongly suggest that mTOR is aberrantly regulated in most AML cells and that rapamycin and analogs, by targeting the clonogenic compartment of the leukemic clone, may be used as new compounds in AML therapy. PMID- 15550489 TI - Liver Kupffer cells rapidly remove red blood cell-derived vesicles from the circulation by scavenger receptors. AB - Previous studies have shown that during the lifespan of red blood cells (RBCs) 20% of hemoglobin is lost by shedding of hemoglobin-containing vesicles. However, the fate of these vesicles is unknown. To study this fate we used a rat model, after having established that rat RBCs lose hemoglobin in the same way as human RBCs, and that RBC-derived vesicles are preferentially labeled by Na2(51) CrO4. Such labeled vesicles were injected into recipient rats. Within 5 minutes, 80% of the radioactivity was cleared from the circulation with a concomitant uptake by the liver of 55% of the injected dose. After 30 minutes, Kupffer cells contained considerable amounts of hemoglobin and were shown to be responsible for 92% of the liver uptake. Vesicle clearance from the blood as well as liver uptake were significantly inhibited by preinjection of the scavenger-receptor ligands polyinosinic acid and phosphatidylserine. We conclude that in rats Kupffer cells rapidly remove RBC-derived vesicles from the circulation, mainly by scavenger receptors. The same mechanism is likely to be responsible for the elimination of human RBC vesicles, thereby constituting an important pathway for the breakdown of RBCs in humans. PMID- 15550490 TI - Molecular profiling of diffuse large B-cell lymphoma identifies robust subtypes including one characterized by host inflammatory response. AB - Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease with recognized variability in clinical outcome, genetic features, and cells of origin. To date, transcriptional profiling has been used to highlight similarities between DLBCL tumor cells and normal B-cell subtypes and associate genes and pathways with unfavorable outcome. To identify robust and highly reproducible DL-BCL subtypes with comprehensive transcriptional signatures, we used a large series of newly diagnosed DLBCLs, whole genome arrays, and multiple clustering methods. Tumors were also analyzed for known common genetic abnormalities in DLBCL. There were 3 discrete subsets of DLBCL-"oxidative phosphorylation," "B-cell receptor/proliferation," and "host response" (HR)-identified characterized using gene set enrichment analysis and confirmed in an independent series. HR tumors had increased expression of T/natural killer cell receptor and activation pathway components, complement cascade members, macrophage/dendritic cell markers, and inflammatory mediators. HR DLB-CLs also contained significantly higher numbers of morphologically distinct CD2+/CD3+ tumor-infiltrating lymphocytes and interdigitating S100+/gamma interferon-induced lysosomal transferase-positive (GILT+) CD1a-/CD123- dendritic cells. The HR cluster shared features of histologically defined T-cell/histiocyte-rich B-cell lymphoma, including fewer genetic abnormalities, younger age at presentation, and frequent splenic and bone marrow involvement. These studies identify tumor microenvironment and host inflammatory response as defining features in DLBCL and suggest rational treatment targets in specific DLBCL subsets. PMID- 15550491 TI - Towards less confusing terminology in reproductive medicine: a counter proposal. AB - Less confusing terminology in reproductive medicine was the goal of a recent proposal the basis of which was to drop the term 'infertility' and replace it with a description. It also contained a prognostic grading system. The present article, arguing against most of these proposals, should not be regarded as nihilism but as constructive criticism to attain more clarity and common understanding. A counter proposal is suggested and includes retention of the term 'infertility', disposal of the terms 'subfertility' and 'sterility' with practical suggestions of when difficulties in conceiving should be investigated and treated. PMID- 15550492 TI - Elective single embryo transfer (eSET) policy in the first three IVF/ICSI treatment cycles. AB - BACKGROUND: Elective single embryo transfer (eSET), applied in the first or second IVF cycle in young patients with good quality embryos, has been demonstrated to lower the twin pregnancy rate, while the overall pregnancy rate is not compromised. It is as yet unclear whether eSET could be the preferred transfer policy in all treatment cycles, or that it should be restricted to the first or first two cycles. METHODS: eSET policy (when two or more embryos were available, at least one of them being of good quality) was offered to patients younger than 38 years in the first three treatment cycles. Retrospectively, treatment cycle outcome was studied. RESULTS: In 326 patients, 586 treatment cycles were performed (326 first, 168 second and 92 third treatment cycles). In 65 cycles (11%), eSET could not be applied because there was either no fertilization, or only one embryo available. In the remaining 521 cycles, eSET was performed in 111 cycles (19%), while in 410 cycles, no good quality embryo was available resulting in the transfer of two embryos (double embryo transfer, DET). No significant differences in ongoing pregnancy rates after transfer of fresh embryos were observed between eSET and DET in the first (both 33%), second (36 and 23%, respectively) and third treatment cycles (20 and 24%, respectively). In significantly more eSET cycles compared to DET cycles, could embryos be frozen. This resulted in a significantly higher cumulative pregnancy rate after eSET compared to DET. CONCLUSIONS: In patients younger than 38 years with at least one top quality embryo, eSET can be the transfer policy of choice in at least the first three treatment cycles, since the pregnancy rates obtained in each treatment cycle are comparable to those after DET. PMID- 15550493 TI - Diurnal changes in FSH-regulatory peptides and their relationship to gonadotrophins in pubertal girls. AB - BACKGROUND: FSH-regulatory peptides participate with GnRH and sex steroids to regulate serum FSH concentrations. We hypothesized that day/night variations in FSH serum concentrations would be associated with diurnal variation in FSH regulatory peptides. METHODS: Blood was obtained every 15 min for 24 h beginning at 08:00 h in eight girls [seven with variations in growth or puberty and one with idiopathic hypogonadotrophic hypogonadism (IHH)] and for 12 h beginning at 20:00 h in 12 additional girls with variant puberty, eight with gonadal dysgenesis or ovarian failure (GD/OF) and one with IHH. Samples across 3 h blocks were pooled for determination of LH, FSH, activin-A, inhibin-B and follistatin 288. RESULTS: LH and FSH concentrations increased from 23:00 to 08:00 h with respect to daytime concentrations in pubertal girls (P<0.005) but only LH increased (P=0.002) in girls with GD/OF. In pubertal girls, inhibin-B declined during the day (P=0.019), reaching a nadir between 17:00 and 22:45 h just prior to the night-time increase in FSH. Follistatin concentrations exhibited diurnal variation (P=0.028), with the greatest concentrations occurring between 05:00 and 11:00 h. Activin-A concentrations declined coincident with the night-time increase in FSH in pubertal girls (P<0.0001) but not in girls with GD/OF. CONCLUSIONS: The directionality of changes in FSH-regulatory proteins supports the notion that FSH-regulatory peptides may contribute to the night-time augmentation of circulating FSH during puberty in girls. PMID- 15550494 TI - Endometriosis affects sperm-endosalpingeal interactions. AB - BACKGROUND: The interaction between spermatozoa and the epithelium of the isthmic region of the uterine tube is thought to be an important part of the mechanisms of sperm transport to the site of fertilization and in preparing them for fertilization. However, it is unclear whether a dysfunction of this mechanism may contribute to subfertility in some individuals. METHODS: The sperm-binding characteristics of the epithelium from the uterine tubes of three groups of women were examined: (i) eight with pelvic endometriosis (not involving the uterine tubes); (ii) five women who had been receiving zoladex injections to control their symptoms; and (iii) as controls 10 women undergoing an elective procedure for benign gynaecological problems but with no other pathology of the reproductive tract. RESULTS: Significantly more spermatozoa bound per unit area to the ampullary epithelium of the uterine tubes taken from women with a previous diagnosis of endometriosis. CONCLUSIONS: These findings suggest that the characteristics of sperm binding to tubal epithelium may be disrupted in women with a gynaecological pathology such as endometriosis. It is suggested that this may have the potential to interfere with the availability of freely motile spermatozoa, of the appropriate physiological status, to take part in fertilization. This may be a newly described mechanism by which endometriosis can cause infertility. PMID- 15550495 TI - Duration of sexual abstinence: epididymal and accessory sex gland secretions and their relationship to sperm motility. AB - BACKGROUND: The data on the association between the abstinence period and sperm motility are conflicting. METHODS: Ejaculates from 422 men assessed for infertility were analysed according to the World Health Organization (WHO) guidelines. Seminal plasma neutral alpha-glucosidase (NAG), prostate-specific antigen (PSA), zinc and fructose were measured. Three groups were defined according to the length of sexual abstinence: G2-3 (2-3 days), G4-5 (4-5 days) and G6-7 (6-7 days). RESULTS: The total percentage of progressively motile spermatozoa was significantly higher in G4-5 compared with G2-3 and G6-7 (medians 55 versus 47 and 42%; P=0.039 and P <0.001, respectively). The percentage of spermatozoa with tail defects was significantly higher in G6-7 compared with G2-3 and G4-5 (medians 14 versus 10 and 10%; P=0.011 and P=0.002, respectively). NAG was significantly lower in G2-3 compared with G4-5 and G6-7 (medians 23 versus 34 and 34 mU/ejaculate; P <0.001 and P=0.001, respectively). The same trend was found regarding zinc (medians 6 versus 8 and 8 mumol/ejaculate; P=0.001 and P=0.005, respectively). CONCLUSIONS: Within the time interval recommended by the WHO (2-7 days), the length of the abstinence period is associated with sperm characteristics and should be taken into consideration when interpreting results of semen analysis. PMID- 15550496 TI - Prospective multicentre randomized controlled trial to evaluate factors influencing the success rate of office diagnostic hysteroscopy. AB - BACKGROUND: Diagnostic hysteroscopy is not widely performed in the office setting, one of the reasons being the discomfort produced by the procedure. This randomized controlled trial was performed to evaluate the effects of instrument diameter, patient parity and surgeon experience on the pain suffered and success rate of the procedure. METHODS: Patients were randomly assigned to undergo office diagnostic hysteroscopy either with 5.0 mm conventional instruments (n=240) or with 3.5 mm mini-instruments (n=240). Procedures were stratified according to patient parity and surgeon's previous experience. The pain experienced during the procedure (0-10), the quality of visualization of the uterine cavity (0-3) and the complications were recorded. The examination was considered successful when the pain score was <4, visualization score was >1 and no complication occurred. RESULTS: Less pain, better visualization and higher success rates were observed with mini-hysteroscopy (P <0.0001, P <0.0001 and P <0.0001, respectively), in patients with vaginal deliveries (P <0.0001, P <0.0001 and P <0.0001, respectively) and in procedures performed by experienced surgeons (P=0.02, P=NS and P=NS, respectively). The effects of patient parity and surgeon experience were no longer important when mini-hysteroscopy was used. CONCLUSIONS: Our data demonstrate the advantages of mini-hysteroscopy and the importance of patient parity and surgeon experience, suggesting that mini-hysteroscopy should always be used, especially for inexperienced surgeons and when difficult access to the uterine cavity is anticipated. They indicate that mini-hysteroscopy can be offered as a first line office diagnostic procedure. PMID- 15550497 TI - ESHRE PGD Consortium data collection IV: May-December 2001. AB - The ESHRE PGD Consortium was formed in 1997 to survey the practice of preimplantation genetic diagnosis (PGD). Since then, three reports have been published giving an overview on PGD from an ever-increasing number of centres and reporting on an increasing number of PGD cycles and pregnancies and babies born after PGD. After these initial influential publications, important shortcomings were identified primarily on the method of data collection, i.e. with Excel spreadsheets, and in the timing of the collection (cycles were collected in a different time frame from pregnancies and babies, making the follow-up of cycles very difficult). This is why the Steering Committee has made a major investment in developing and implementing a new database in FileMaker Pro 6. It was also decided that cycles would be collected from one calendar year, as well as the pregnancies and babies ensuing from that particular calendar year. This gave us the opportunity to take a closer look at the data collected earlier, and to attempt to improve their quality. This is a report on the corrected data from the first three data collections (I-III) as well as the result of the last data collection (IV) that was completely carried out using the new database. PMID- 15550498 TI - DNA sequence variations of the entire anti-Mullerian hormone (AMH) gene promoter and AMH protein expression in patients with the Mayer-Rokitanski-Kuster-Hauser syndrome. AB - BACKGROUND: The etiology of the Mayer-Rokitanski-Kuster-Hauser (MRKH) syndrome, where congenitally the Mullerian ducts fail to develop into the uterus, cervix and upper vagina, along with other malformations, is unresolved. Anti-Mullerian hormone (AMH) signal transduction inducing the degradation of Mullerian ducts in males is implicated in the MRKH syndrome. This study examined if DNA sequence variations are responsible for the activation of AMH and aberrant hormone levels in MRKH patients. METHODS: The entire AMH promoter and 3' regulatory elements of the constitutively expressed splicing factor SF3a2 were sequenced in 30 MRKH patients and genotyped in 48 control individuals using matrix-assisted laser desorption/ionization-time-of-flight mass spectronomy. Ovarian AMH promoter function was correlated with protein expression in plasma and peritoneal fluid of MRKH patients. RESULTS: Of six identified AMH promoter variations, two at positions -639 (SP1-binding site) and -210 [steroidogenic factor (SF)1-binding site] were homozygote in 73% of patients, and 69% of control individuals, destroying the SP1-binding site. AMH protein levels in plasma and peritoneal fluid from patients were equivalent to control individuals, however in three patients plasma levels were abnormally high. CONCLUSIONS: AMH is an important indicator for ovarian function. AMH promoter sequence variations or the previously proposed SF3a2-AMH fusion co-transcripts cannot be responsible for aberrant AMH expression leading to Mullerian duct degradation. PMID- 15550499 TI - Reproductive ageing and ovarian function: is the early follicular phase FSH rise necessary to maintain adequate secretory function in older ovulatory women? AB - BACKGROUND: Serum FSH elevations and decreases in inhibin B have been consistently demonstrated in the early follicular phase of cycles in women of advanced reproductive age. However, secretory products of the dominant follicle (estradiol and inhibin A) in the serum of older ovulatory women are maintained at levels similar to those of their younger counterparts. The goal of this investigation was to determine if ovarian secretory capacity is dependent on relative FSH levels and if basal measures of ovarian reserve reflect ovarian secretory capacity. METHODS: We administered equivalent low, but effective doses of recombinant FSH for 5 days to a group of older subjects (40-45 years, n=9) and younger controls (20-25 years, n=10) after pituitary suppression with a GnRH agonist. Outcome measures included follicular development as determined by serial transvaginal ultrasound examinations and serum levels of estradiol, inhibin A and inhibin B. RESULTS: Serum levels of estradiol and inhibin A were not statistically different between the two groups, while the number of large follicles formed was greater in the younger subjects. Basal parameters of ovarian reserve were not significantly correlated with ovarian secretory capacity, but did correlate with the number of follicles recruited in response to low-dose FSH. CONCLUSIONS: By providing equivalent serum levels of FSH in older and younger reproductive aged women, this study demonstrates that the secretory capacity of recruited follicles is maintained in older reproductive aged women. PMID- 15550500 TI - Intrauterine insemination versus Fallopian tube sperm perfusion in non-tubal subfertility: a systematic review based on a Cochrane review. AB - BACKGROUND: The objective of this review was to compare the efficacy of Fallopian tube sperm perfusion (FSP) with intrauterine insemination (IUI) in the treatment of non-tubal subfertility. METHODS: The principles of the Cochrane Menstrual Disorders and Subfertility Group were employed. Only randomized controlled studies comparing FSP with IUI were included in this review. The main outcome measures included live birth rates and pregnancy rates per couple. RESULTS: Twenty-eight studies were found performing the comparison of interest. Overall six studies involving 474 couples were included in the meta-analysis. One study only assessed live birth rates, which resulted in no difference in outcome between FSP and IUI [odds ratio (OR) 1.17, 95% confidence interval (CI) 0.39 3.53]. The results in pregnancy rate per couple revealed no statistically significant difference between FSP and IUI (OR 1.76, 95% CI 0.77-4.05). Subgroup analysis revealed that couples suffering from unexplained subfertility clearly benefit from FSP over IUI (OR 2.88, 95% CI 1.73-4.78). Excluding studies which used the Foley catheter for tubal perfusion resulted in a significant difference favouring FSP for all indications (OR 2.42, 95% CI 1.54-3.80). CONCLUSIONS: There is firm evidence that FSP gives rise to higher pregnancy rates in couples with unexplained subfertility and should therefore be advised in these couples. For other indications FSP has not been proven more effective compared with IUI. Results showed that the Foley catheter might not be effective for FSP. Future research should focus on comparing different types of catheters. PMID- 15550501 TI - Pain relief using electro-acupuncture for oocyte retrieval. PMID- 15550502 TI - Seasonal appearance and seasonal disappearance of menstrual function. PMID- 15550503 TI - Thyroid hormone-dependent gene expression in differentiated embryonic stem cells and embryonal carcinoma cells: identification of novel thyroid hormone target genes by deoxyribonucleic acid microarray analysis. AB - T3 is required for normal early development, but relatively few T3-responsive target genes have been identified. In general, in vitro stem cell differentiation techniques stimulate a wide range of developmental programs, including thyroid hormone receptor (TR) pathways. We developed several in vitro stem cell models to more specifically identify TR-mediated gene expression in early development. We found that embryonic carcinoma (EC) cells have reduced T3 nuclear binding capacity and only modestly express the known T3 target genes, neurogranin (RC3) and Ca2+/calmodulin-dependent protein kinase IV (CaMKIV), in response to T3. Full T3 induction in transient transfection of EC cells was restored with cotransfection of a TR expression vector. We, therefore, performed gene expression profiles in wild-type embryonic stem (ES) cells compared with expression in cells with deficient (EC) or mutant TR (TRalpha P398H mutant ES cells), to identify T3 target genes. T3 stimulation of wild-type ES cells altered mRNA expression of 610 known genes (26% of those studied), although only approximately 60 genes (1%) met criteria for direct T3 stimulation based on the magnitude of induction and requirement for the presence of TR. We selected five candidate T3 target genes, neurexophilin 2, spermatid perinuclear RNA-binding protein (SPNR), kallikrein-binding protein (KBP), prostate-specific membrane antigen (PSMA), and synaptotagmin II, for more detailed study. T3 responsiveness of these genes was evaluated in both in vitro endogenous gene expression and in vivo mouse model systems. These genes identified in a novel stem cell system, including those induced and repressed in response to T3, may mediate thyroid hormone actions in early development. PMID- 15550504 TI - Eplerenone blocks nongenomic effects of aldosterone on the Na+/H+ exchanger, intracellular Ca2+ levels, and vasoconstriction in mesenteric resistance vessels. AB - There is increasing evidence for rapid nongenomic effects of aldosterone. Aldosterone has been demonstrated to alter intracellular pH and calcium in isolated cells. However, few studies have correlated these effects with aldosterone-mediated physiological responses. Therefore, we studied rapid effects of aldosterone on vascular reactivity, intracellular Ca2+, and pH in resistance vessels. Furthermore, we explored whether the new antimineralocorticoid drug eplerenone could effectively block nongenomic aldosterone-mediated effects. The vasoconstrictor action of aldosterone was examined directly by determining the diameter of small resistance mesenteric vessels (160-200 microm resting diameter), simultaneously with intracellular pH or Ca2+. Aldosterone (10 nm) caused a rapid constriction of resistance vessels (8.1% +/- 1.0% reduction in the diameter below control conditions, P < 0.05). Aldosterone potentiated phenylephrine-mediated constriction in small and large mesenteric vessels. Aldosterone induced a rapid increase of intracellular Ca2+ and cellular alkalinization. Vasoconstrictor action of aldosterone and nongenomic effects on the sodium-proton exchanger (NHE1) activity or intracellular Ca2+ responses was abolished by eplerenone. The vasoconstrictor response of aldosterone was related to phosphatidylinositol 3-kinase (PI3-K): the hormone decreased protein kinase B phosphorylation; pharmacological inhibition of PI3-K (10 microm LY294002 or 1 microm wortmannin) increased arterial contractility. Inhibitors of ERK 1/2 phosphorylation (15 microm PD98059) had no effect on aldosterone-mediated vasoconstriction. Inhibition of protein kinase C with 1 microm bi sindolylmaleimide I and/or inhibition of NHE1 with 100 microm amiloride abolished aldosterone vasoconstrictor action of resistance mesenteric arteries. We conclude that aldosterone-mediated increase in vascular tone is related to a nongenomic mechanism that involves protein kinase C, PI3-K, and NHE1 activity. Eplerenone is an effective blocker of nongenomic effects of aldosterone in vascular tissue. PMID- 15550505 TI - Autocrine activation of the local insulin-like growth factor I system is up regulated by estrogen receptor (ER)-independent estrogen actions and accounts for decreased ER expression in type 2 diabetic mesangial cells. AB - Autocrine activation of the IGF-I system in mesangial cells (MC) promotes glomerular scarring in a model of type 1 diabetes. Although estrogens protect against progressive nondiabetic glomerulosclerosis (GS), women with diabetes seem to loose the estrogen-mediated protection against cardiovascular disease. However, little is known about the local IGF-I system and its interactions with estrogens in the pathogenesis of type 2 diabetic GS. Therefore, we examined db/db B6 (db/db) mice, a model of type 2 diabetes and diabetic GS. The IGF-I system was activated in the glomeruli and MC of female diabetic db/db mice, but not in nondiabetic db/+ littermates. We found increased IGF-I receptor (IGFR) expression and activation, including activation of MAPK. Surprisingly, estrogens, via an estrogen receptor (ER)-independent mechanism(s), increased IGFR expression, IGFR and insulin receptor substrate phosphorylation, and extracellular signal regulated kinase activation in db/db MC. In contrast, ER expression was decreased in MC and glomeruli of db/db mice. Treatment with a neutralizing antibody to IGF I or the MAPK inhibitor PD98059 increased ER expression and transcriptional activity. This suggests that the local prosclerotic IGF-I system is activated in type 2 diabetes and diminishes ER-mediated protection against GS. Although estrogens may stimulate protective ER signaling, they also activate the IGF-I system via ER-independent mechanisms in db/db MC. The later estrogen effects appear to outweigh the antisclerotic effects of ER activation. This may in part account for loss of estrogen protection against the progression of diabetic GS in women with type 2 diabetes. PMID- 15550506 TI - alpha1A-adrenoceptors activate glucose uptake in L6 muscle cells through a phospholipase C-, phosphatidylinositol-3 kinase-, and atypical protein kinase C dependent pathway. AB - The role of alpha1-adrenoceptor activation on glucose uptake in L6 cells was investigated. The alpha1-adrenoceptor agonist phenylephrine [pEC50 (-log10 EC50), 5.27 +/- 0.30] or cirazoline (pEC50, 5.00 +/- 0.23) increased glucose uptake in a concentration-dependent manner, as did insulin (pEC50, 7.16 +/- 0.21). The alpha2 adrenoceptor agonist clonidine was without any stimulatory effect on glucose uptake. The stimulatory effect of cirazoline was inhibited by the alpha1 adrenoceptor antagonist prazosin, but not by the beta-adrenoceptor antagonist propranolol. RT-PCR showed that the alpha1A-adrenoceptor was the sole alpha1 adrenoceptor subtype expressed in L6 cells. Cirazoline- or insulin-mediated glucose uptake was inhibited by the phosphatidylinositol-3 kinase inhibitor LY294002, suggesting a possible interaction between the alpha1-adrenoceptor and insulin pathways. Cirazoline or insulin stimulated phosphatidylinositol-3 kinase activity, but alpha1-adrenoceptor activation did not phosphorylate Akt. Both cirazoline- and insulin-mediated glucose uptake were inhibited by protein kinase C (PKC), phospholipase C, and p38 kinase inhibitors, but not by Erk1/2 inhibitors (despite both treatments being able to phosphorylate Erk1/2). Insulin and cirazoline were able to activate and phosphorylate p38 kinase. The phorbol ester 12-O-tetradecanoylphorbol-13-acetate and the calcium ionophore A23187 produced significant increases in glucose uptake, indicating roles for PKC and calcium in glucose uptake. Down-regulation of conventional PKC isoforms inhibited glucose uptake mediated by 12-O-tetradecanoylphorbol-13-acetate, but not by insulin or cirazoline. This study demonstrates that alpha1-adrenoceptors mediate increases in glucose uptake in L6 muscle cells. This effect appears to be related to activation of phospholipase C, phosphatidylinositol-3 kinase, p38 kinase, and PKC. PMID- 15550507 TI - Reduced adipose glucocorticoid reactivation and increased hepatic glucocorticoid clearance as an early adaptation to high-fat feeding in Wistar rats. AB - Altered peripheral glucocorticoid metabolism may be important in the pathogenesis of obesity in humans and animal models. Genetically obese Zucker rats, Lep/ob mice, and obese humans exhibit increased regeneration of active glucocorticoids selectively in adipose tissue by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD-1) and increased glucocorticoid clearance by hepatic A-ring reductases. We have examined whether dietary obesity in rats induces the same changes in glucocorticoid metabolism. Male Wistar rats were weaned onto high-fat (HF; 45% kcal from fat) or control (10% fat) diets. After 3 wk, HF rats showed no differences in weight but were glucose intolerant, had lower 11beta-HSD-1 activity in liver (3.8 +/- 0.2 vs. 4.9 +/- 0.2 pmol product/min.mg protein; P <0.01), sc fat (0.03 +/- 0.01 vs. 0.09 +/- 0.01 pmol product/min.mg protein; P <0.01), and omental fat (0.02 +/- 0.001 vs. 0.03 +/- 0.003 pmol/ product/min.mg protein; P <0.05) and higher hepatic 5beta-reductase activity (0.26 +/- 0.05 vs. 0.10 +/- 0.007 pmol product/min.mg protein; P <0.05). After 20 wk, HF rats were obese, hyperglycemic, and hyperinsulinemic, but differences in 11beta-HSD-1 and 5beta-reductase activities were no longer apparent. Mature male rats given HF diets for 24 or 72 h showed increased hepatic 5beta-reductase activity and a trend for decreased sc adipose 11beta-HSD-1 activity. Dietary obesity is not accompanied by the changes in 11beta-HSD-1 and 5beta-reductase expression and activity observed in genetically obese rodents. Acute exposure to HF diet alters glucocorticoid metabolism, predicting lower hepatic and adipose intracellular glucocorticoid concentrations, which may be a key mechanism protecting against the metabolic complications of obesity. PMID- 15550508 TI - Direct effect of melatonin on Syrian hamster testes: melatonin subtype 1a receptors, inhibition of androgen production, and interaction with the local corticotropin-releasing hormone system. AB - Besides the hypothalamus and pituitary, melatonin action at the testicular level has been recently suggested. Therefore, we investigated in the Syrian hamster, a well-characterized seasonal breeder, melatonin action on Leydig cells, testicular expression of melatonergic receptors, and possible interactions between melatonin receptors and the previously identified testicular serotoninergic and CRH systems. In isolated Leydig cells from active testes of adult hamsters kept in a long-day (14 h light, 10 h dark) photoperiod and from regressed testes of adult animals exposed to a short-day photoperiod during 16 wk (6 h light, 18 h dark), melatonin significantly reduced human chorionic gonadotropin-stimulated production of cAMP and the main androgens: testosterone and androstane 3alpha,17beta-diol, respectively, and decreased the expression of steroidogenic acute regulatory protein, P450 side chain cleavage, 3beta-hydroxysteroid dehydrogenase and 17beta-hydroxysteroid dehydrogenase. In Leydig cells exposed to a short-day photoperiod during 16 wk, melatonin stimulated the conversion of testosterone into 5alpha-reduced androgens by inducing 5alpha-reductase isoform 1, and controlled androstane-3alpha,17beta-diol production by inhibiting 3alpha hydroxysteroid dehydrogenase expression. Melatonin subtype (mel1a) receptors were detected in Leydig cells. Although the local serotonin system did not mediate melatonin action on androgen production, melatonergic effect on steroidogenesis involved the interaction between mel1a receptors and the inhibitory CRH system. Moreover, melatonin significantly increased CRH mRNA levels and production in hamster Leydig cells expressing CRH subtype 1 receptors. Our studies indicate that melatonin may act as a local inhibitor of human chorionic gonadotropin stimulated cAMP and androgen production through mel1a receptors, down-regulation of steroidogenic acute regulatory protein, and key steroidogenic enzymes expression and its interaction with the local CRH system. PMID- 15550509 TI - Overexpression of gly56/gly80/gly81-mutant insulin-like growth factor-binding protein-3 in transgenic mice. AB - IGF-independent effects of IGF-binding protein-3 (IGFBP-3) have been demonstrated in vitro; however, the physiological significance of these effects in vivo is unclear. We generated two transgenic (Tg) mouse strains that overexpress a human Gly56/Gly80/Gly81-mutant IGFBP-3 cDNA. This mutant has a markedly reduced affinity for the IGFs, but retains the IGF-independent effects. Serum levels of mutant IGFBP-3 were 156 +/- 12 and 400 +/- 24 ng/ml in hemizygous mice of strains 5005 and 5012, respectively. When Tg and wild-type mice were compared, there was no reduction in birth weight, litter size, or postnatal growth. Despite differences in transgene expression in various tissues, relative organ weight was similar in Tg and wild-type mice, with exception of brain, where a modest reduction in brain weight was observed in the high-expressing 5012 lineage. There was also a significant reduction in proliferating cell nuclear antigen-staining cells observed in the periventricular region of the developing brain in embryonic d 18 Tg embryos. In the higher expressing 5012 Tg strain, IGF-I and murine IGFBP 3 levels, marker of GH action were increased. Furthermore, there was a positive correlation between mutant IGFBP-3 levels and IGF-I levels and between mutant IGFBP-3 levels and murine IGFBP-3 (P = 0.002 and P < 0.001, respectively). These data indicate that overexpression of mutant IGFBP-3 is not associated with growth retardation. The higher levels of IGF-I and murine IGFBP-3 in the 5012 Tg strain suggest that the growth inhibitory effect of mutant IGFBP-3 may be compensated for by other mechanisms. PMID- 15550510 TI - Short-term in vivo inhibition of insulin receptor substrate-1 expression leads to insulin resistance, hyperinsulinemia, and increased adiposity. AB - Insulin receptor substrate-1 (IRS-1) has an important role as an early intermediary between the insulin and IGF receptors and downstream molecules that participate in insulin and IGF-I signal transduction. Here we employed an antisense oligonucleotide (IRS-1AS) to inhibit whole-body expression of IRS-1 in vivo and evaluate the consequences of short-term inhibition of IRS-1 in Wistar rats. Four days of treatment with IRS-1AS reduced the expression of IRS-1 by 80, 75, and 65% (P < 0.05) in liver, skeletal muscle, and adipose tissue, respectively. This was accompanied by a 40% (P < 0.05) reduction in the constant of glucose decay during an insulin tolerance test, a 78% (P < 0.05) reduction in glucose consumption during a hyperinsulinemic-euglycemic clamp, and a 90% (P < 0.05) increase in basal plasma insulin level. The metabolic effects produced by IRS-1AS were accompanied by a significant reduction in insulin-induced [Ser (473)] Akt phosphorylation in liver (85%, P < 0.05), skeletal muscle (40%, P < 0.05), and adipose tissue (85%, P < 0.05) and a significant reduction in insulin induced tyrosine phosphorylation of ERK in liver (20%, P < 0.05) and skeletal muscle (30%, P < 0.05). However, insulin-induced tyrosine phosphorylation of ERK was significantly increased (60%, P < 0.05) in adipose tissue of IRS-1AS-treated rats. In rats treated with IRS-1AS for 8 d, a 100% increase (P < 0.05) in relative epididymal fat weight and a 120% (P < 0.05) increase in nuclear expression of peroxisome proliferator-activated receptor-gamma were observed. Thus, acute inhibition of IRS-1 expression in rats leads to insulin resistance accompanied by activation of a growth-related pathway exclusively in white adipose tissue. PMID- 15550511 TI - Daily variations in type II iodothyronine deiodinase activity in the rat brain as controlled by the biological clock. AB - Type II deiodinase (D2) plays a key role in regulating thyroid hormone-dependent processes in, among others, the central nervous system (CNS) by accelerating the intracellular conversion of T4 into active T3. Just like the well-known daily rhythm of the hormones of the hypothalamo-pituitary-thyroid axis, D2 activity also appears to show daily variations. However, the mechanisms involved in generating these daily variations, especially in the CNS, are not known. Therefore, we decided to investigate the role the master biological clock, located in the hypothalamus, plays with respect to D2 activity in the rat CNS as well as the role of one of its main hormonal outputs, i.e. plasma corticosterone. D2 activity showed a significant daily rhythm in the pineal and pituitary gland as well as hypothalamic and cortical brain tissue, albeit with a different timing of its acrophase in the different tissues. Ablation of the biological clock abolished the daily variations of D2 activity in all four tissues studied. The main effect of the knockout of the suprachiasmatic nuclei (SCN) was a reduction of nocturnal peak levels in D2 activity. Moreover, contrary to previous observations in SCN-intact animals, in SCN-lesioned animals, the decreased levels of D2 activity are accompanied by decreased plasma levels of the thyroid hormones, suggesting that the SCN separately stimulates D2 activity as well as the hypothalamo-pituitary-thyroid axis. PMID- 15550512 TI - Cyclic adenosine 3',5'-monophosphate (cAMP) enhances cAMP-responsive element binding (CREB) protein phosphorylation and phospho-CREB interaction with the mouse steroidogenic acute regulatory protein gene promoter. AB - Steroidogenic acute regulatory protein (StAR) transcription is regulated through cAMP-protein kinase A-dependent mechanisms that involve multiple transcription factors including the cAMP-responsive element binding protein (CREB) family members. Classically, binding of phosphorylated CREB to cis-acting cAMP responsive elements (5'-TGACGTCA-3') within target gene promoters leads to recruitment of the coactivator CREB binding protein (CBP). Herein we examined the extent of CREB family member phosphorylation on protein-DNA interactions and CBP recruitment with the StAR promoter. Immunoblot analysis revealed that CREB, cAMP responsive element modulator (CREM), and activating transcription factor (ATF)-1 are expressed in MA-10 mouse Leydig tumor cells, yet only CREB and ATF-1 are phosphorylated. (Bu)2cAMP treatment of MA-10 cells increased CREB phosphorylation approximately 2.3-fold within 30 min but did not change total nuclear CREB expression levels. Using DNA-affinity chromatography, we now show that CREB and ATF-1, but not CREM, interact with the StAR promoter, and this interaction is dependent on the activator protein-1 (AP-1) cis-acting element within the cAMP responsive region. In addition, (Bu)2cAMP-treatment increased phosphorylated CREB (P-CREB) association with the StAR promoter but did not influence total CREB interaction. In vivo chromatin immunoprecipitation assays demonstrated CREB binding to the StAR proximal promoter is independent of (Bu)2cAMP-treatment, confirming our in vitro analysis. However, (Bu)2cAMP-treatment increased P-CREB and CBP interaction with the StAR promoter, demonstrating for the first time the physical role of P-CREB:DNA interactions in CBP recruitment to the StAR proximal promoter. PMID- 15550513 TI - Autocrine regulation of insulin secretion in human ejaculated spermatozoa. AB - A striking feature of insulin expression is its almost complete restriction to beta-cells of the pancreatic islet in normal mammals. Here we show that insulin is expressed in and secreted from human ejaculated spermatozoa. Both insulin transcript and protein were detected. In addition, the large differences in insulin secretion, assessed by RIA, between noncapacitated and capacitated sperm suggest a role for insulin in capacitation. Insulin had an oscillatory secretory pattern involving glucose dose-dependent increases and significant decreases during the blockage of an insulin autocrine effect. It appears that the effect of glucose on the fertilizing ability of sperm is mediated by glucose metabolism through the pentose phosphate pathway. Then we evaluated the autocrine effect of sperm insulin on glucose metabolism by studying the activity of glucose-6 phosphate dehydrogenase, the key rate-limiting enzyme in the pentose phosphate pathway. The simultaneous decrease in both insulin release and glucose-6 phosphate dehydrogenase activity induced by blocking the autocrine insulin effect with three different procedures (blockage of insulin release by nifedipine, immune neutralization of the released insulin by antiinsulin serum, and blockage of an insulin intracellular effector such as phosphotidylinositol 3-kinase by wortmannin) strongly suggests a physiological role of sperm insulin on these two events. Insulin secretion by spermatozoa may provide an autocrine regulation of glucose metabolism based on their energetic needs independent of systemic insulin. In conclusion, these data open a new area of study in male reproduction. PMID- 15550515 TI - Estrogen receptor-alpha mediates estrogen facilitation of baroreflex heart rate responses in conscious mice. AB - Estrogen facilitates baroreflex heart rate responses evoked by intravenous infusion of ANG II and phenylephrine (PE) in ovariectomized female mice. The present study aims to identify the estrogen receptor subtype involved in mediating these effects of estrogen. Baroreflex responses to PE, ANG II, and sodium nitroprusside (SNP) were tested in intact and ovariectomized estrogen receptor-alpha knockout (ERalphaKO) with (OvxE+) or without (OvxE-) estrogen replacement. Wild-type (WT) females homozygous for the ERalpha(+/+) were used as controls. Basal mean arterial pressures (MAP) and heart rates were comparable in all the groups except the ERalphaKO-OvxE+ mice. This group had significantly smaller resting MAP, suggesting an effect of estrogen on resting vascular tone possibly mediated by the ERbeta subtype. Unlike the WT females, estrogen did not facilitate baroreflex heart rate responses to either PE or ANG II in the ERalphaKO-OvxE+ mice. The slope of the line relating baroreflex heart rate decreases with increases in MAP evoked by PE was comparable in ERalphaKO-OvxE- ( 6.97 +/- 1.4 beats.min(-1).mmHg(-1)) and ERalphaKO-OvxE+ (-6.18 +/- 1.3) mice. Likewise, the slope of the baroreflex bradycardic responses to ANG II was similar in ERalphaKO-OvxE- (-3.87 +/- 0.5) and ERalphaKO-OvxE+(-2.60 +/- 0.5) females. Data suggest that estrogen facilitation of baroreflex responses to PE and ANG II is predominantly mediated by ERalpha subtype. A second important observation in the present study is that the slope of ANG II-induced baroreflex bradycardia is significantly blunted compared with PE in the intact as well as the ERalphaKO OvxE+ females. We have previously reported that this ANG II-mediated blunting of cardiac baroreflexes is observed only in WT males and not in ovariectomized WT females independent of their estrogen replacement status. The present data suggest that in females lacking ERalpha, ANG II causes blunting of cardiac baroreflexes similar to males and may be indicative of a direct modulatory effect of the ERalpha on those central mechanisms involved in ANG II-induced resetting of cardiac baroreflexes. These observations suggest an important role for ERalpha subtype in the central modulation of baroreflex responses. Lastly, estrogen did not significantly affect reflex tachycardic responses to SNP in both WT and ERalphaKO mice. PMID- 15550516 TI - Dynamics of flow velocities in endocardial and epicardial coronary arterioles. AB - The subendocardium is the most vulnerable area of the left ventricle to the effects of hypoperfusion and ischemia. Despite this well-acknowledged observation, the mechanisms underlying this susceptibility are not elucidated, although numerous explanations including differences in transmural distribution of hemodynamics, metabolism, and wall stresses have been proposed. Our goal was to make dynamic measurements of endocardial and epicardial flow velocities, which reflect hemodynamic and wall stresses, to approach this problem. We measured blood flow velocities in subendocardial and subepicardial coronary arterioles of in vivo beating canine hearts using a high-speed, charge-coupled device, intravital videomicroscope with a rod-probe lens. Subendocardial flow was characterized by remarkable systolic flow-velocity reversal (systolic slosh ratio, 84%; measurable velocity of retrograde flow, faster than -40 mm/s), which contrasted to predominant forward-flow velocity during systole in the subepicardial arterioles (systolic slosh ratio, 25%; maximum velocity, approximately -20 mm/s; P < 0.0005 and 0.05 vs. subendocardial arterioles, respectively). We speculate that this retrograde flow is "wasteful," because this volume must be refilled during the subsequent diastole, which thereby detracts from the net perfusion as well as the time for perfusion. Accordingly, we also believe that the retrograde systolic blood flow contributes to the vulnerability of the subendocardium to ischemia. PMID- 15550514 TI - Insulin-like growth factor-binding protein-5 induces a gender-related decrease in bone mineral density in transgenic mice. AB - IGF-binding protein-5 (IGFBP-5) is abundant in serum and bone during normal skeletal development, but levels decrease in osteoporosis. Studies have shown that IGFBP-5 stimulates markers of bone formation by potentiating IGF actions and by IGF-independent actions. To test the hypothesis that IGFBP-5 promotes the acquisition of bone mineral density (BMD), we generated transgenic (Tg) mice overexpressing Igfbp5 using a cytomegalovirus enhancer and beta-actin promoter (CMV/betaA). Tg animals showed an increase in serum IGFBP-5 concentrations by 7.7 to 3.5-fold at 3-8 wk of age, respectively. Concentrations were 6-49% higher for males compared with females in both wild-type and Tg mice. Surprisingly, BMD decreased in a gender-dependent manner, with Tg male adults affected more severely than Tg females (31.3% vs. 19.2% reduction, respectively, compared with wild-type mice, assessed by dual energy x-ray absorptiometry). Significant gender differences in BMD were confirmed by peripheral quantitative computed tomography. Histomorphometry revealed that although the bone formation rate and mineralizing surface at the periosteum decreased in Tg mice, they increased at the endosteum, suggesting opposing effects of IGFBP-5 on periosteal and endosteal osteoblasts (by altering proliferation or survival). These findings differ from previous observations in Igf1- and Igf2-null animals. In conclusion, IGFBP-5 has a significant influence on BMD acquisition and maintenance that is dependent on gender and age. The phenotype of Igfbp5 mice cannot be explained solely by IGF inhibition; thus, this study provides the first in vivo evidence, by genetic manipulation, for IGF-independent actions of IGFBP-5 in bone function. These findings have implications for the gender-biased progression of osteoporosis. PMID- 15550518 TI - Haptoglobin expression and activity during coronary collateralization. AB - Coronary collateral development relies on the coordinated secretion of growth factors. However, alone they are insufficient for permanent collateral growth. We utilized proteomics to identify other important proteins in the extracellular environment that could facilitate collateralization. Chronically instrumented dogs developed coronary collaterals by the repetitive occlusion method. Subendocardial (0.19 +/- 0.04, 0.27 +/- 0.06, 0.48 +/- 0.10, and 0.81 +/- 0.11 ml x min(-1) x g(-1) on days 1, 7, 14, and 21, respectively) and subepicardial (0.14 +/- 0.01, 0.36 +/- 0.06, 0.51 +/- 0.07, and 0.71 +/- 0.08 ml x min(-1) x g(-1) on days 1, 7, 14, and 21, respectively) blood flow increased in animals subjected to repetitive occlusion. Sham animals exhibited no changes in blood flow. Myocardial interstitial fluid (MIF) from both groups was analyzed by two-dimensional electrophoresis with matrix-assisted laser desorption/ionization time-of-flight identification. The acute-phase protein haptoglobin was identified in the group subjected to repetitive occlusion. ELISA of MIF showed haptoglobin to be elevated at all time points of collateral development compared with sham, with maximal production on day 7. Purified haptoglobin dose dependently stimulated endothelial cells to form tubes and vascular smooth muscle cells to migrate. Purified haptoglobin did not stimulate proliferation of either cell type. The relative contribution of haptoglobin to the chemotactic properties of MIF was tested using a neutralizing antibody. Neutralized MIF could not stimulate smooth muscle cells to migrate at any time during collateral development. Endothelial cell tube formation was inhibited after the midpoint of collateralization. Therefore, the acute-phase protein haptoglobin plays a critical role during coronary collateralization. PMID- 15550517 TI - mitoKATP channel activation in the postanoxic developing heart protects E-C coupling via NO-, ROS-, and PKC-dependent pathways. AB - Whereas previous studies have shown that opening of the mitochondrial ATP sensitive K(+) (mitoK(ATP)) channel protects the adult heart against ischemia reperfusion injury, it remains to be established whether this mechanism also operates in the developing heart. Isolated spontaneously beating hearts from 4 day-old chick embryos were subjected to 30 min of anoxia followed by 60 min of reoxygenation. The chrono-, dromo-, and inotropic disturbances, as well as alterations of the electromechanical delay (EMD), reflecting excitation contraction (E-C) coupling, were investigated. Production of reactive oxygen species (ROS) in the ventricle was determined using the intracellular fluorescent probe 2',7'-dichlorofluorescin (DCFH). Effects of the specific mitoK(ATP) channel opener diazoxide (Diazo, 50 microM) or the blocker 5-hydroxydecanoate (5-HD, 500 microM), the nitric oxide synthase (NOS) inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 50 microM), the antioxidant N-(2-mercaptopropionyl)glycine (MPG, 1 mM), and the PKC inhibitor chelerythrine (Chel, 5 microM) on oxidative stress and postanoxic functional recovery were determined. Under normoxia, the baseline parameters were not altered by any of these pharmacological agents, alone or in combination. During the first 20 min of postanoxic reoxygenation, Diazo doubled the peak of ROS production and, interestingly, accelerated recovery of ventricular EMD and the PR interval. Diazo-induced ROS production was suppressed by 5-HD, MPG, or L-NAME, but not by Chel. Protection of ventricular EMD by Diazo was abolished by 5-HD, MPG, L-NAME, or Chel, whereas protection of the PR interval was abolished by L-NAME exclusively. Thus pharmacological opening of the mitoK(ATP) channel selectively improves postanoxic recovery of cell-to-cell communication and ventricular E-C coupling. Although the NO-, ROS-, and PKC dependent pathways also seem to be involved in this cardioprotection, their interrelation in the developing heart can differ markedly from that in the adult myocardium. PMID- 15550519 TI - Reduced oxygen supply explains the negative force-frequency relation and the positive inotropic effect of adenosine in buffer-perfused hearts. AB - In isolated rat hearts perfused with HEPES and red blood cell-enriched buffers, we examined changes in left ventricular pressure induced by increases in heart rate or infusion of adenosine to investigate whether the negative force-frequency relation and the positive inotropic effect of adenosine are related to an inadequate oxygen supply provided by crystalloid perfusates. Hearts perfused with HEPES buffer at a constant flow demonstrated a negative force-frequency relation, whereas hearts perfused with red blood cell-enriched buffer exhibited a positive force-frequency relation. In contrast, HEPES buffer-perfused hearts showed a concentration-dependent increase in left ventricular systolic pressure [EC50 = 7.0 +/- 1.2 nM, maximal effect (Emax) = 104 +/- 2 and 84 +/- 2 mmHg at 0.1 microM and baseline, respectively] in response to adenosine, whereas hearts perfused with red blood cell-enriched buffer showed no change in left ventricular pressure. The positive inotropic effect of adenosine correlated with the simultaneous reduction in heart rate (r = 0.67, P < 0.01; EC50 = 3.8 +/- 1.4 nM, baseline 228 +/- 21 beats/min to a minimum of 183 +/- 22 beats/min at 0.1 microM) and was abolished in isolated hearts paced to suppress the adenosine-induced bradycardia. In conclusion, these results indicate that the negative force frequency relation and the positive inotropic effect of adenosine in the isolated rat heart are related to myocardial hypoxia, rather than functional peculiarities of the rat heart. PMID- 15550520 TI - Functional effects of C-type natriuretic peptide and nitric oxide are attenuated in hypertrophic myocytes from pressure-overloaded mouse hearts. AB - Increases in the myocardial level of cGMP usually exert negative inotropic effects in the mammalian hearts. We tested the hypothesis that the negative functional effects caused by nitric oxide (NO) or C-type natriuretic peptide (CNP) through cGMP would be blunted in hypertrophied cardiac myocytes. Contractile function, guanylyl cyclase activity, cGMP-dependent protein phosphorylation, and calcium transients were assessed in ventricular myocytes from aortic stenosis-induced hypertrophic and age-matched control mice. Basal percentage shortening was similar in control and hypertrophic myocytes. S-nitroso N-acetyl-penicillamine (SNAP, an NO donor, 10(-6) and 10(-5) M) or CNP (10(-8) and 10(-7) M) reduced percentage shortening in both groups, but their effects were blunted in hypertrophic myocytes. Maximal rates of shortening and relaxation were depressed at the basal level, and both reagents had attenuated effects in hypertrophy. Similar results were also found after treatment with guanylin and carbon monoxide, other stimulators of particulate, and soluble guanylyl cyclase, respectively. Guanylyl cyclase activity was not significantly changed in hypertrophy. Addition of Rp-8-[(4-chlorophenyl)thio]-cGMPS triethylamine (an inhibitor of cGMP-dependent protein kinase, 5 x 10(-6) M) blocked SNAP or the effect of CNP in control mice but not in hypertrophy, indicating the cGMP dependent kinase (PKG) may not mediate the actions of cGMP induced by NO or CNP in the hypertrophic state. Calcium transients after SNAP or CNP were not significantly changed in hypertrophy. These results suggest that in hypertrophied mice, diminished effects of NO or CNP on ventricular myocyte contraction are not due to changes in guanylyl cyclase activity. The data also indicated that PKG mediated pathways were diminished in hypertrophied myocardium, contributing to blunted effects. PMID- 15550521 TI - Direct measurement of transmural laminar architecture in the anterolateral wall of the ovine left ventricle: new implications for wall thickening mechanics. AB - Laminar, or sheet, architecture of the left ventricle (LV) is a structural basis for normal systolic and diastolic LV dynamics, but transmural sheet orientations remain incompletely characterized. We directly measured the transmural distribution of sheet angles in the ovine anterolateral LV wall. Ten Dorsett hybrid sheep hearts were perfusion fixed in situ with 5% buffered glutaraldehyde at end diastole and stored in 10% formalin. Transmural blocks of myocardial tissue were excised, with the edges cut parallel to local circumferential, longitudinal, and radial axes, and sliced into 1-mm-thick sections parallel to the epicardial tangent plane from epicardium to endocardium. Mean fiber directions were determined in each section from five measurements of fiber angles. Each section was then cut transverse to the fiber direction, and five sheet angles (beta) were measured and averaged. Mean fiber angles progressed nearly linearly from -41 degrees (SD 11) at the epicardium to +42 degrees (SD 16) at the endocardium. Two families of sheets were identified at approximately +45 degrees (beta(+)) and -45 degrees (beta(-)). In the lateral region (n = 5), near the epicardium, sheets belonged to the beta(+) family; in the midwall, to the beta(-) family; and near the endocardium, to the beta(+) family. This pattern was reversed in the basal anterior region (n = 4). Sheets were uniformly beta(-) over the anterior papillary muscle (n = 2). These direct measurements of sheet angles reveal, for the first time, alternating transmural families of predominant sheet angles. This may have important implications in understanding wall mechanics in the normal and the failing heart. PMID- 15550523 TI - Fatty acid oxidation and its impact on response of spontaneously hypertensive rat hearts to an adrenergic stress: benefits of a medium-chain fatty acid. AB - The spontaneously hypertensive rat (SHR) is a model of cardiomyopathy characterized by a restricted use of exogenous long-chain fatty acid (LCFA) for energy production. The aims of the present study were to document the functional and metabolic response of the SHR heart under conditions of increased energy demand and the effects of a medium-chain fatty acid (MCFA; octanoate) supplementation in this situation. Hearts were perfused ex vivo in a working mode with physiological concentrations of substrates and hormones and subjected to an adrenergic stimulation (epinephrine, 10 microM). (13)C-labeled substrates were used to assess substrate selection for energy production. Compared with control Wistar rat hearts, SHR hearts showed an impaired response to the adrenergic stimulation as reflected by 1) a smaller increase in contractility and developed pressure, 2) a faster decline in the aortic flow, and 3) greater cardiac tissue damage (lactate dehydrogenase release: 1,577 +/- 118 vs. 825 +/- 44 mU/min, P < 0.01). At the metabolic level, SHR hearts presented 1) a reduced exogenous LCFA contribution to the citric acid cycle flux (16 +/- 1 vs. 44 +/- 4%, P < 0.001) and an enhanced contribution of endogenous substrates (20 +/- 4 vs. 1 +/- 4%, P < 0.01); and 2) an increased lactate production from glycolysis, with a greater lactate-to-pyruvate production ratio. Addition of 0.2 mM octanoate reduced lactate dehydrogenase release (1,145 +/- 155 vs. 1,890 +/- 89 mU/min, P < 0.001) and increased exogenous fatty acid contribution to energy metabolism (23.7 +/- 1.3 vs. 15.8 +/- 0.8%, P < 0.01), which was accompanied by an equivalent decrease in unlabeled endogenous substrate contribution, possibly triglycerides (11.6 +/- 1.5 vs. 19.0 +/- 1.2%, P < 0.01). Taken altogether, these results demonstrate that the SHR heart shows an impaired capacity to withstand an acute adrenergic stress, which can be improved by increasing the contribution of exogenous fatty acid oxidation to energy production by MCFA supplementation. PMID- 15550522 TI - Endothelial dysfunction and hypercontractility of vascular myocytes are ameliorated by fluvastatin in obese Zucker rats. AB - To study the mechanisms of vascular dysfunction in diabetes mellitus, we examined the responses of the aorta to adrenomedullin (AM) and ANG II in obese Zucker (OZ), lean Zucker (LZ), and OZ rats administered fluvastatin (OZ + Flu). AM induced endothelium-dependent vasorelaxation was impaired in OZ rats compared with LZ rats, and fluvastatin restored AM-induced, endothelium-dependent vasorelaxation (%Deltatension at 10(-7) mol/l AM; LZ, -85.1 +/- 3.1%; OZ, -50.7 +/- 2.5%; OZ + Flu, -75.6 +/- 2.7%). Expression of endothelial nitric oxide synthase (eNOS) and Akt phosphorylation in response to AM (10(-7) mol/l) were also diminished in OZ rats. Fluvastatin restored the eNOS expression and Akt phosphorylation [eNOS expression (relative intensity): LZ, 2.3 +/- 0.4; OZ, 1.0 +/- 0.2; OZ + Flu, 1.8 +/- 0.3; Akt phosphorylation (relative intensity): LZ, 2.3 +/- 0.2; OZ, 1.0 +/- 0.3; OZ + Flu, 1.9 +/- 0.2]. ANG II-induced vasoconstriction was enhanced in the aortic rings of OZ rats compared with LZ rats, and this enhanced vasoconstriction was partially normalized by fluvastatin and was abolished when the aorta of OZ rats was preincubated with the Rho kinase inhibitor Y-27632. GTPgammaS-induced contraction of permeabilized aortic smooth muscle cells, which is an indicator of the Rho-dependent Ca(2+) sensitization of contraction, was enhanced in OZ rats compared with LZ rats, and this enhanced contraction was suppressed in OZ + Flu rats. These results suggested that endothelium-dependent vasorelaxation was impaired, Ca(2+) sensitization of contraction was augmented in blood vessels of OZ rats and that fluvastatin restored vascular function by activating the Akt-dependent pathway and inhibiting the Rho-dependent pathway. PMID- 15550524 TI - Hypertrophic cardiomyopathy-related beta-myosin mutations cause highly variable calcium sensitivity with functional imbalances among individual muscle cells. AB - Disease-causing mutations in cardiac myosin heavy chain (beta-MHC) are identified in about one-third of families with hypertrophic cardiomyopathy (HCM). The effect of myosin mutations on calcium sensitivity of the myofilaments, however, is largely unknown. Because normal and mutant cardiac MHC are also expressed in slow twitch skeletal muscle, which is more easily accessible and less subject to the adaptive responses seen in myocardium, we compared the calcium sensitivity (pCa(50)) and the steepness of force-pCa relations (cooperativity) of single soleus muscle fibers from healthy individuals and from HCM patients of three families with selected myosin mutations. Fibers with the Arg723Gly and Arg719Trp mutations showed a decrease in mean pCa(50), whereas those with the Ile736Thr mutation showed slightly increased mean pCa(50) with higher active forces at low calcium concentrations and residual active force even under relaxing conditions. In addition, there was a marked variability in pCa(50) between individual fibers carrying the same mutation ranging from an almost normal response to highly significant differences that were not observed in controls. While changes in mean pCa(50) may suggest specific pharmacological treatment (e.g., calcium antagonists), the observed large functional variability among individual muscle cells might negate such selective treatment. More importantly, the variability in pCa(50) from fiber to fiber is likely to cause imbalances in force generation and be the primary cause for contractile dysfunction and development of disarray in the myocardium. PMID- 15550525 TI - Iron chelation and a free radical scavenger suppress angiotensin II-induced upregulation of TGF-beta1 in the heart. AB - Long-term administration of angiotensin II causes myocardial loss and cardiac fibrosis. We previously found iron deposition in the heart of the angiotensin II infused rat, which may promote angiotensin II-induced cardiac damage. In the present study, we have investigated whether an iron chelator (deferoxamine) and a free radical scavenger (T-0970) affect the angiotensin II-induced upregulation of transforming growth factor-beta1 (TGF-beta1). Angiotensin II infusion for 7 days caused a robust increase in TGF-beta1 mRNA expression in vascular smooth muscle cells, myofibroblast-like cells, and migrated monocytes/macrophages. T-0970 and deferoxamine suppressed the upregulation of TGF-beta1 mRNA and reduced the extent of cardiac fibrosis in the heart of rats treated with angiotensin II. These agents blocked the angiotensin II-induced upregulation of heme oxygenase-1, a potent oxidative and cellular stress-responsive gene, but they did not significantly affect systolic blood pressure or plasma levels of aldosterone. In addition, T-0970 and deferoxamine suppressed the angiotensin II-induced upregulation of monocyte chemoattractant protein-1 in the heart. These results collectively suggest that iron and the iron-mediated generation of reactive oxygen species may contribute to angiotensin II-induced upregulation of profibrotic and proinflammatory genes, such as TGF-beta1 and monocyte chemoattractant protein-1. PMID- 15550526 TI - Engineering skeletal myoblasts: roles of three-dimensional culture and electrical stimulation. AB - Immature skeletal muscle cells, or myoblasts, have been used in cellular cardiomyoplasty in attempts to regenerate cardiac muscle tissue by injection of cells into damaged myocardium. In some studies, muscle tissue within myoblast implant sites may be morphologically similar to cardiac muscle. We hypothesized that identifiable aspects of the cardiac milieu may contribute to growth and development of implanted myoblasts in vivo. To test this hypothesis, we designed a novel in vitro system to mimic some aspects of the electrical and biochemical environment of native myocardium. This system enabled us to separate the three dimensional (3-D) electrical and biochemical signals that may be involved in myoblast proliferation and plasticity. Myoblasts were grown on 3-D polyglycolic acid mesh scaffolds under control conditions, in the presence of cardiac-like electrical current fluxes, or in the presence of culture medium that had been conditioned by mature cardiomyocytes. Cardiac-like electrical current fluxes caused increased myoblast number in 3-D culture, as determined by DNA assay. The increase in cell number was due to increased cellular proliferation and not differences in apoptosis, as determined by proliferating cell nuclear antigen and TdT-mediated dUTP nick-end labeling. Cardiomyocyte-conditioned medium also significantly increased myoblast proliferation. Expression of transcription factors governing differentiation along skeletal or cardiac lineages was evaluated by immunoblotting. Although these assays are qualitative, no changes in differentiation state along skeletal or cardiac lineages were observed in response to electrical current fluxes. Furthermore, from these experiments, conditioned medium did not appear to alter the differentiation state of skeletal myoblasts. Hence, cardiac milieu appears to stimulate proliferation but does not affect differentiation of skeletal myoblasts. PMID- 15550527 TI - Normal distribution of ventricular pressure-volume area of arrhythmic beats under atrial fibrillation in canine heart. AB - We previously found the frequency distribution of the left ventricular (LV) effective afterload elastance (E(a)) of arrhythmic beats to be nonnormal or non Gaussian in contrast to the normal distribution of the LV end-systolic elastance (E(max)) in canine in situ LVs during electrically induced atrial fibrillation (AF). These two mechanical variables determine the total mechanical energy [systolic pressure-volume area (PVA)] generated by LV contraction when the LV end diastolic volume is given on a per-beat basis. PVA and E(max) are the two key determinants of the LV O(2) consumption per beat. In the present study, we analyzed the frequency distribution of PVA during AF by its chi(2), significance level, skewness, and kurtosis and compared them with those of other major cardiodynamic variables including E(a) and E(max). We assumed the volume intercept (V(0)) of the end-systolic pressure-volume relation needed for E(max) determination to be stable during arrhythmia. We found that PVA distributed much more normally than E(a) and slightly more so than E(max) during AF. We compared the chi(2), significance level, skewness, and kurtosis of all the complex terms of the PVA formula. We found that the complexity of the PVA formula attenuated the effect of the considerably nonnormal distribution of E(a) on the distribution of PVA along the central limit theorem. We conclude that mean (SD) of PVA can reliably characterize the distribution of PVA of arrhythmic beats during AF, at least in canine hearts. PMID- 15550528 TI - Adaptation to mechanical load determines shape and properties of heart and circulation: the CircAdapt model. AB - With circulatory pathology, patient-specific simulation of hemodynamics is required to minimize invasiveness for diagnosis, treatment planning, and followup. We investigated the advantages of a smart combination of often already known hemodynamic principles. The CircAdapt model was designed to simulate beat to-beat dynamics of the four-chamber heart with systemic and pulmonary circulation while incorporating a realistic relation between pressure-volume load and tissue mechanics and adaptation of tissues to mechanical load. Adaptation was modeled by rules, where a locally sensed signal results in a local action of the tissue. The applied rules were as follows: For blood vessel walls, 1) flow shear stress dilates the wall and 2) tensile stress thickens the wall; for myocardial tissue, 3) strain dilates the wall material, 4) larger maximum sarcomere length increases contractility, and 5) contractility increases wall mass. The circulation was composed of active and passive compliances and inertias. A realistic circulation developed by self-structuring through adaptation provided mean levels of systemic pressure and flow. Ability to simulate a wide variety of patient-specific circumstances was demonstrated by application of the same adaptation rules to the conditions of fetal circulation followed by a switch to the newborn circulation around birth. It was concluded that a few adaptation rules, directed to normalize mechanical load of the tissue, were sufficient to develop and maintain a realistic circulation automatically. Adaptation rules appear to be the key to reduce dramatically the number of input parameters for simulating circulation dynamics. The model may be used to simulate circulation pathology and to predict effects of treatment. PMID- 15550529 TI - Long-term infusion of Met5-enkephalin fails to protect murine hearts against ischemia-reperfusion injury. AB - Recently, we reported that exogenous administration of Met(5)-enkephalin (ME) for 24 h reduces infarct size after ischemia-reperfusion in rabbits. In the present study, we tested whether ME-induced cardioprotection is exhibited in murine hearts and whether chronic infusion of this peptide can render hearts tolerant to ischemia. Barbiturate-anesthetized open-chest mice (C57BL/6J) were subjected to regional myocardial ischemia-reperfusion (45 min of occlusion and 20 min of reperfusion). Mice received saline vehicle or ME for 24 h or 2 wk before undergoing regional myocardial ischemia-reperfusion or for 24 h followed by a 24 h delay before regional myocardial ischemia-reperfusion. Infarct size was measured with propidium iodide and is expressed as a percentage of the area at risk. Infarcts were smaller after infusion of ME for 24 h than with vehicle control: 49.2 +/- 9.0% vs. 22.2 +/- 3.2% (P < 0.01). In contrast, administration of ME for 2 wk failed to elicit cardioprotection: 36.5 +/- 9.1% and 41.4 +/- 8.2% for control and ME, respectively (P = not significant). When a 24-h delay was imposed between the end of drug treatment and the onset of the ischemic insult, cardioprotection was lost: 38.5 +/- 6.1% and 42.8 +/- 6.6% for control and ME, respectively (P = not significant). Chronic sustained exogenous infusion of the endogenously produced opioid peptide ME is associated with loss of the cardioprotection that is observed with 24 h of infusion. Furthermore, in this in vivo murine model, ME failed to induce delayed tolerance to myocardial ischemia reperfusion. PMID- 15550530 TI - Differential effects of exercise on aortic mitochondria. AB - Routine exercise is widely recognized as cardioprotective. Exercise induces a variety of effects within the cardiovasculature, including decreased mitochondrial damage and improved aerobic capacity. It has been generally thought that the transient increase in oxidative stress associated with exercise initiates cardioprotective processes. Somewhat paradoxically, increased oxidative stress associated with cardiovascular disease (CVD) risk factors is thought to play an important role in the promotion and development of CVD. Hence, it is possible that CVD risk factors that increase oxidative stress (e.g., hypercholesterolemia) may modulate the cardioprotective effects of exercise. In this regard, the interaction between CVD risk factors and exercise on atherosclerotic lesion development and basal oxidant load is less defined. To determine the influence of preexistent hypercholesterolemia on cardioprotective effects of exercise, atherosclerotic lesion formation, oxidant load, mitochondrial damage, protein nitration (3-nitrotyrosine levels), and mitochondrial enzyme activities were determined in aortic tissues from normocholesterolemic (C57 control) and hypercholesterolemic [apoliprotein E deficient (apoE(-/-))] mice after 16 wk of regular exercise. In normocholesterolemic mice, regular exercise was associated with decreased mitochondrial damage and oxidant load and increased SOD2 and adenine nucleotide translocator activities. Exercise did not decrease endogenous oxidant load and mitochondrial damage in hypercholesterolemic mice and did not reduce atherosclerotic lesion development. These data are consistent with the notion that CVD risk factors associated with increased oxidative stress can alter the benefits of exercise and that mitochondrial damage appears to be correlated with the cardiovascular effects of exercise. PMID- 15550531 TI - Tolerance and short term efficacy of rituximab in 43 patients with systemic autoimmune diseases. AB - OBJECTIVE: To assess the tolerance and efficacy of rituximab in patients with various autoimmune diseases seen in daily rheumatological practice. METHODS: 866 rheumatology and internal medicine practitioners were contacted by e-mail to obtain the files of patients treated with rituximab for systemic autoimmune diseases. Patients with lymphoma were analysed if the evolution of the autoimmune disease could be evaluated. RESULTS: In all, 43 of 49 cases could be analysed, including 14 with rheumatoid arthritis (RA), 13 with systemic lupus erythematosus (SLE), six with primary Sjogren's syndrome (pSS), five with systemic vasculitis, and five with other autoimmune diseases. Rituximab was prescribed for lymphoma in two patients with RA and two with pSS. In the 39 other cases, rituximab was given because of the refractory character of the autoimmune disease. The mean follow up period was 8.3 months (range 2 to 26). There were 11 adverse events in 10 patients and treatment had to be discontinued in six. Efficacy was observed in 30 patients (70%): RA 11, SLE 9, pSS 5, vasculitis 2, antisynthetase syndromes 2, sarcoidosis 1. The mean decrease in corticosteroid intake was 9.5 mg/d (range 0 to 50) in responders. Seven patients experienced relapse after mean 8.1 months (5 to 15). Three patients died because of refractory autoimmune disease. CONCLUSIONS: Despite absence of marketing authorisation, rituximab is used to treat various refractory autoimmune diseases in daily rheumatological practice. This study showed good tolerance and short term clinical efficacy, with marked corticosteroid reduction in patients with SLE, pSS, vasculitis, and polymyositis. PMID- 15550532 TI - Male microchimerism in women with systemic sclerosis and healthy women who have never given birth to a son. AB - BACKGROUND: Male DNA or cells are often used to measure microchimerism in a woman. In studies of autoimmune diseases male microchimerism is most often attributed to the previous birth of a son. OBJECTIVE: To determine the frequency of male microchimerism in healthy women or women with systemic sclerosis who had never given birth to a son. METHODS: Real time quantitative polymerase chain reaction targeting the Y chromosome specific sequence DYS14 was employed to test DNA extracted from peripheral blood mononuclear cells of 26 women with systemic sclerosis and 23 healthy women who had never given birth to a son. RESULTS: are expressed as the genome equivalent number of male cells per million host cells (gEq/mil).Results: Male DNA was found in 15% of women with systemic sclerosis (range 0 to 23.7 gEq/mil) and in 13% of healthy women (range 0 to 5.1 gEq/mil). Although two women with male DNA had an induced abortion, most had no history of spontaneous or induced abortion (either systemic sclerosis or healthy). CONCLUSIONS: Microchimerism with male DNA can be found in the circulation of women who have never given birth to a son. Thus sources other than a male birth must be considered when male DNA is used to measure microchimerism. Although other studies are needed, there was no apparent difference in women with systemic sclerosis and healthy women. Possible sources of male DNA include unrecognised male pregnancy or unrecognised male twin, an older male sibling with transfer through the maternal circulation, or sexual intercourse alone. PMID- 15550533 TI - Cost effectiveness of adalimumab in the treatment of patients with moderate to severe rheumatoid arthritis in Sweden. AB - BACKGROUND: Societal decision makers increasingly emphasise their need for evidence based economic analyses to make reimbursement decisions. OBJECTIVE: To analyse the cost utility of adalimumab, on both incremental cost and incremental quality adjusted life years (QALYs), versus traditional disease modifying antirheumatic drugs and the other tumour necrosis factor (TNF) antagonists suitable for submission to the Swedish LFN (Pharmaceutical Benefit Board). METHODS: Swedish unit costs and treatment guidelines from a lifetime perspective were implemented. A mathematical model, incorporating data from seven trials, simulated the experiences of 10 000 hypothetical patients with moderate to severe rheumatoid arthritis (RA). The primary outcome measure-QALYs-was derived from utility values calculated from a relationship between the Health Assessment Questionnaire (HAQ) Disability Index (DI) and Health Utility Index-III (HUI-3) from adalimumab trial results. The model followed the progression of HAQ-DI through a number of treatments in a sequence accounting for mortality, drug and monitoring costs, and other direct costs. RESULTS: When using ACR50 as a response threshold for determining successful treatment, adalimumab plus methotrexate showed the greatest number of QALYs gained (2.3 from one study and 2.1 from the pooled results of two trials). The etanercept plus methotrexate strategy yielded QALY gains similar to the pooled adalimumab results. Except for the infliximab strategy, the costs results were between 35 000 and 42 000, a range normally considered cost effective in other European countries. CONCLUSION: Adalimumab appears to be cost effective for the treatment of moderate to severe RA. The results suggest that adalimumab is at least as cost effective as other TNF antagonists. PMID- 15550534 TI - Reactive arthritis attributable to Shigella infection: a clinical and epidemiological nationwide study. AB - OBJECTIVES: To study the incidence and clinical picture of Shigella associated reactive arthritis (ReA) and the arthritogenicity of various Shigella species in the population. METHODS: A questionnaire on enteric and extraintestinal, especially musculoskeletal, symptoms was sent to 278 consecutive patients with Shigella positive stool culture and to 597 controls. Analysis of self reported musculoskeletal symptoms was supplemented with clinical examination of those subjects with recent symptoms. RESULTS: Of the patients, 14/211 (7%) had ReA, and a further 4/211 (2%) other reactive musculoskeletal symptoms (tendonitis, enthesopathy, or bursitis). Of the 14 patients with ReA, all adults, 10 had S sonnei, three S flexneri, and one S dysenteriae infection. HLA-B27 was positive in 36% of the patients with ReA. One control subject had ReA. In the patients with Shigella infection, the odds ratio for developing ReA was 16.2 (95% confidence interval 2.1 to 123.9), p = 0.001. CONCLUSIONS: ReA occurred in 7% of patients after Shigella infection, with an annual incidence of 1.3/1 000 000 in Finland. Besides S flexneri, S sonnei and S dysenteriae can also trigger ReA. PMID- 15550535 TI - Ultrasound guided synovial biopsy using portal and forceps. AB - OBJECTIVE: To describe a new method for taking a synovial biopsy specimen under ultrasound guidance using portal and forceps. METHODS: Percutaneous ultrasound guided biopsy was performed for 37 patients with mono- or polyarthritis as outpatients. A portal to a planned area was built using a needle, guiding wire, and dilators, through which forceps could be inserted and samples taken. Biopsy samples were taken from small and large joints, bursae, and tendon sheaths. RESULTS: Representative synovial tissue in adequate amounts for histopathological evaluation was obtained in 33/37 cases--a success rate of 89%. The biopsy procedures were well tolerated, but one complication of skin infection was encountered. CONCLUSION: The new method of synovium biopsy under ultrasound guidance using sheath introducer set and flexible forceps can be performed on most joints and even bursae and tendon sheaths. The method gives sufficient samples for clinical work in most cases, but further work is needed before accepting this promising technique for scientific purposes. PMID- 15550536 TI - Bioluminescent imaging of a marking transgene and correction of Fabry mice by neonatal injection of recombinant lentiviral vectors. AB - Successful therapy for many inherited disorders could be improved if the intervention were initiated early. This is especially true for lysosomal storage disorders. Earlier intervention may allow metabolic correction to occur before lipid buildup has irreversible consequences and/or before the immune system mounts limiting responses. We have been developing gene therapy to treat lysosomal storage disorders, especially Fabry disease. We describe studies directed toward metabolic correction in neonatal animals mediated by recombinant lentiviral vectors. To develop this method, we first injected a marking lentiviral vector that engineers expression of luciferase into the temporal vein of recipient neonatal animals. The use of a cooled charged-coupled device camera allowed us to track transgene expression over time in live animals. We observed intense luciferase expression in many tissues, including the brain, that did not diminish over 24 weeks. Next, we injected neonatal Fabry mice a single time with a therapeutic lentiviral vector engineered to express human alpha-galactosidase A. The injection procedure was well tolerated. We observed increased plasma levels of alpha-galactosidase A activity starting at our first plasma collection point (4 weeks). Levels of alpha-galactosidase A activity were found to be significantly elevated in many tissues even after 28 weeks. No immune response was observed against the corrective transgene product. Increased levels of enzyme activity also led to significant reduction of globotriaosylceramide in the liver, spleen, and heart. This approach provides a method to treat lysosomal storage disorders and other disorders before destructive manifestations occur. PMID- 15550537 TI - Prediction of the structural motifs of sandwich proteins. AB - We investigate the supersecondary structure of a large group of proteins, the so called sandwich proteins. The analysis of a large number of such proteins has led us to propose a set of rules that can be used to predict the possible arrangements of strands in the two beta-sheets forming a given sandwich structure. These rules imply the existence of certain invariant supersecondary substructures common to all sandwich proteins. Furthermore, they dramatically restrict the number of permissible arrangements. For example, whereas for proteins consisting of three strands in each beta-sheet 180 possible strand arrangements exist a priori, our rules imply that only 15 of them are permissible. Five of these predicted arrangements describe all currently known sandwich proteins with six strands. PMID- 15550538 TI - Anaerobic growth of Methanosarcina acetivorans C2A on carbon monoxide: an unusual way of life for a methanogenic archaeon. AB - All methanogenic Archaea examined to date rely on methanogenesis as their sole means of energy conservation. Among these are ones that use carbon monoxide as a growth substrate, producing methane via a pathway that involves hydrogen as an intermediate. To further examine the role of hydrogen in this process, we tested the ability of Methanosarcina acetivorans C2A, a metabolically versatile methanogen devoid of significant hydrogen metabolism, to use CO as a growth substrate. M. acetivorans grew on CO to high cell densities (approximately 1 x 10(8) per ml) with a doubling time of approximately 24 h. Surprisingly, acetate and formate, rather than methane, were the major metabolic end products as shown by 13C NMR studies and enzymatic analysis of culture supernatants. Methane formation surpassed acetate/formate formation only when the cultures entered stationary growth phase, strongly suggesting that M. acetivorans conserves energy by means of this acetogenic and formigenic process. Resting cell experiments showed that methane production decreased linearly with increasing CO partial pressures, consistent with inhibition of methanogenesis by CO. Transposon-induced M. acetivorans mutants with lesions in the operon encoding phosphotransacetylase and acetate kinase failed to use either acetate or CO as growth substrates, indicating that these enzymes are required for both aceticlastic methanogenesis and carboxidotrophic acetogenesis. These findings greatly extend our concept of energy conservation and metabolic versatility in the methanogenic Archaea. PMID- 15550539 TI - A highly specific L-galactose-1-phosphate phosphatase on the path to ascorbate biosynthesis. AB - Ascorbate is a critical compound in plants and animals. Humans are unable to synthesize ascorbate, and their main source of this essential vitamin are plants. However, the pathway of synthesis in plants is yet to be established, and several unknown enzymes are only postulated to exist. We describe a specific L-galactose 1-phosphate (L-gal-1-P) phosphatase that we partially purified from young kiwifruit (Actinidia deliciosa) berries. The enzyme had a native molecular mass of approximately 65 kDa, was completely dependent on Mg2+ for activity and was very specific in its ability to hydrolyze L-gal-1-P. The activity had a pH optimum of 7.0, a K(-M(L-gal-1-P) of 20-40 microM and a Ka(Mg2+) of 0.2 mM. The activity was inhibited by Mg2+ at concentrations >2 mM. The enzyme from Arabidopsis thaliana shoots showed similar properties to the kiwifruit enzyme. The Arabidopsis thaliana enzyme preparation was digested with trypsin, and proteins present were identified by using liquid chromatography-MS. One of 24 proteins present in our preparation was an Arabidopsis thaliana protein, At3g02870, annotated myo-inositol-1-phosphate phosphatase in GenBank, that matched the characteristics of the purified l-gal-1-phosphate phosphatase. We then expressed a kiwifruit homologue of this gene in Escherichia coli and found that it showed 14-fold higher maximum velocity for l-gal-1-P than myo-inositol-1 P. The expressed enzyme showed very similar properties to the enzyme purified from kiwifruit and Arabidopsis, except that its KM(L-gal-1-P) and Ka(Mg2+) were higher in the expressed enzyme. The data are discussed in terms of the pathway to ascorbate biosynthesis in plants. PMID- 15550540 TI - The organization of orientation and spatial frequency in primary visual cortex. AB - Many studies have demonstrated that the primary visual cortex contains multiple functional maps of visual properties (e.g., ocular dominance, orientation preference, and spatial-frequency preference), but as yet no consistent picture has emerged as to how these maps are related to one another. Three divergent, prior optical-imaging studies of spatial frequency are reanalyzed and critiqued in this article. Evidence is presented that a nonstimulus-specific response biased the interpretation of results in previous studies. In addition to reexamining four prior cat experiments, we carried out one new experiment. Through the use of different methods and a careful removal of the nonspecific response, we are led in all instances to a unique view of cortical organization for spatial-frequency preference. In particular, we find little apparent evidence for a columnar organization for spatial frequency. The response recorded by each image pixel may be viewed as arising from an admixture of low- and high-spatial frequency populations. For most pixels, the ratio of these populations is 1:1. PMID- 15550541 TI - Discovery of functional noncoding elements by digital analysis of chromatin structure. AB - We developed a quantitative methodology, digital analysis of chromatin structure (DACS), for high-throughput, automated mapping of DNase I-hypersensitive sites and associated cis-regulatory sequences in the human and other complex genomes. We used 19/20-bp genomic DNA tags to localize individual DNase I cutting events in nuclear chromatin and produced approximately 257,000 tags from erythroid cells. Tags were mapped to the human genome, and a quantitative algorithm was applied to discriminate statistically significant clusters of independent DNase I cutting events. We show that such clusters identify both known regulatory sequences and previously unrecognized functional elements across the genome. We used in silico simulation to demonstrate that DACS is capable of efficient and accurate localization of the majority of DNase I-hypersensitive sites in the human genome without requiring an independent validation step. A unique feature of DACS is that it permits unbiased evaluation of the chromatin state of regulatory sequences from widely separated genomic loci. We found surprisingly large differences in the accessibility of distant regulatory sequences, suggesting the existence of a hierarchy of nuclear organization that escapes detection by conventional chromatin assays. PMID- 15550542 TI - dsu functions in a MYO5A-independent pathway to suppress the coat color of dilute mice. AB - MYO5A is a major actin-based vesicle transport motor that binds to one of its cargos, the melanosome, by means of a RAB27A/MLPH receptor. When one of the members of this receptor-motor complex is mutated, the melanosomes clump in the perinuclear region of the melanocyte and are transferred unevenly to the developing hair, leading to a dilution of coat color. Mutation of a fourth gene, dilute suppressor (dsu), suppresses this coat color dilution. MYO5A is required for the peripheral accumulation of melanosomes in melanocytes, but its role in melanosome transfer to neighboring keratinocytes and the hair is unknown. Here, we show that MYO5A is nonessential for melanosome transfer, although pigment incorporation into the hair in MYO5A-deficient mice is uneven, probably due to the clumping of melanosomes that occurs in the perinuclear region of mutant melanocytes. We also show that dsu is caused by a loss-of-function mutation in a unique vertebrate-specific protein that appears to function in an MYO5A independent pathway to alter pigment incorporation into the hair. Therefore, dsu identifies a unique protein involved in pigmentation of the mammalian hair. PMID- 15550543 TI - Activation of the complement system generates antibacterial peptides. AB - The complement system represents an evolutionary old and significant part of the innate immune system involved in protection against invading microorganisms. Here, we show that the anaphylatoxin C3a and its inactivated derivative C3a desArg are antibacterial, demonstrating a previously unknown direct antimicrobial effect of complement activation. The C3a peptide, as well as functional epitopes in the sequence, efficiently killed the Gram-negative bacteria Escherichia coli, Pseudomonas aeruginosa, and the Gram-positive Enterococcus faecalis. In mice, a C3a-derived peptide suppressed infection by Gram-positive Streptococcus pyogenes bacteria. Fluorescence and electron microscopy demonstrated that C3a binds to and induces breaks in bacterial membranes. C3a was also found to induce membrane leakage of liposomes. These findings provide an interesting link between the complement system and antimicrobial peptides, which are two important branches of innate immunity. PMID- 15550544 TI - Human and murine inhibitory natural killer cell receptors transfer from natural killer cells to target cells. AB - Intercellular transfer of proteins across the immunological synapse is emerging as a common outcome of immune surveillance. We previously reported that target cell MHC class I protein transfers onto natural killer (NK) cells expressing cognate killer Ig-like receptors (KIRs). We now show that, for both murine and human cells, target cells expressing inhibitory MHC class I ligands acquire cognate inhibitory NK receptors. Other cell-surface proteins, but not a cytoplasmic dye, also transferred from human NK cells to target cells across an inhibitory immunological synapse. The number of KIRs acquired from NK cells correlated with the level of expression of cognate MHC class I protein on target cells. Treatment with cytoskeletal inhibitors demonstrated that the target-cell cytoskeleton influences intercellular transfer of proteins in both directions. In contrast to constitutively expressed KIRs, a fraction of acquired KIRs could be removed by mild acid wash, demonstrating a difference between some of the acquired KIRs and constitutively expressed KIRs. An accumulation of phosphotyrosine at the location of the transferred KIRs implies a signaling capacity for NK cell proteins transferred to target cells. Thus, intercellular protein transfer between immune cells is bidirectional and could facilitate new aspects of immune cell communication. PMID- 15550545 TI - Differential nitros(yl)ation of blood and tissue constituents during glyceryl trinitrate biotransformation in vivo. AB - Nitric oxide (NO)-derived products may modify tissue constituents, forming S- and N-nitroso adducts and metal nitrosyls implicated in NO signaling. Nitrovasodilator drugs have been in widespread use for more than a century, yet their biotransformation pathways to NO and their effects as NO donors across tissues remain ill defined. By using a metabonomics approach (termed "NObonomics") for detailing the global NO-related metabolism of the cornerstone nitrovasodilator, glyceryl trinitrate (GTN; 0.1-100 mg/kg), in the rat in vivo, we find that GTN biotransformation elicits extensive tissue nitros(yl)ation throughout all major organ systems. The corresponding reaction products remained detectable hours after administration, and vascular tissue was not a major nitros(yl)ation site. Extensive heart and liver modifications involved both S- and N-nitrosation, and RBC S-nitrosothiol formation emerged as a sensitive indicator of organic nitrate metabolism. The dynamics of GTN-derived oxidative NO metabolites in blood did not reflect the nitros(yl)ation patterns in the circulation or in tissues, casting doubt on the usefulness of plasma nitrite/nitrate as an index of NO/NO-donor biodynamics. Target-tissue NO metabolites varied in amount and type with GTN dose, suggesting a dose-sensitive shift in the prevailing routes of GTN biotransformation ("metabolic shunting") from thiol nitrosation to heme nitrosylation. We further demonstrate that GTN induced nitros(yl)ation is modulated by a complex, tissue-selective interplay of enzyme-catalyzed pathways. These findings provide insight into the global in vivo metabolism of GTN at pharmacologically relevant doses and offer an additional experimental paradigm for the NObonomic analysis of NO-donor metabolism and signaling. PMID- 15550546 TI - Running on water: Three-dimensional force generation by basilisk lizards. AB - Water provides a unique challenge for legged locomotion because it readily yields to any applied force. Previous studies have shown that static stability during locomotion is possible only when the center of mass remains within a theoretical region of stability. Running across a highly yielding surface could move the center of mass beyond the edges of the region of stability, potentially leading to tripping or falling. Yet basilisk lizards are proficient water runners, regularly dashing across bodies of water to evade predators. We present here direct measurements of time-averaged force produced by juvenile plumed basilisk lizards (Basiliscus plumifrons) while running across water. By using digital particle image velocimetry to visualize fluid flow induced by foot movement, we show that sufficient support force is generated for a lizard to run across water and that novel strategies are also required to run across a highly yielding surface. Juvenile basilisk lizards produce greatest support and propulsive forces during the first half of the step, when the foot moves primarily vertically downwards into the water; they also produce large transverse reaction forces that change from medial (79% body weight) to lateral (37% body weight) throughout the step. These forces may act to dynamically stabilize the lizards during water running. Our results give insight into the mechanics of how basilisk lizards run across water and, on a broader scale, provide a conceptual basis for how locomotor surface properties can challenge established rules for the mechanics of legged locomotion. PMID- 15550547 TI - Ca2+ activity at GABAB receptors constitutively promotes metabotropic glutamate signaling in the absence of GABA. AB - Type B gamma-aminobutyric acid receptor (GABABR) is a G protein-coupled receptor that regulates neurotransmitter release and neuronal excitability throughout the brain. In various neurons, GABABRs are concentrated at excitatory synapses. Although these receptors are assumed to respond to GABA spillover from neighboring inhibitory synapses, their function is not fully understood. Here we show a previously undescribed function of GABABR exerted independent of GABA. In cerebellar Purkinje cells, interaction of GABABR with extracellular Ca2+ (Ca(2+)o) leads to a constitutive increase in the glutamate sensitivity of metabotropic glutamate receptor 1 (mGluR1). mGluR1 sensitization is clearly mediated by GABABR because it is absent in GABABR1 subunit-knockout cells. However, the mGluR1 sensitization does not require G(i/o) proteins that mediate the GABABR's classical functions. Moreover, coimmunoprecipitation reveals complex formation between GABABR and mGluR1 in the cerebellum. These findings demonstrate that GABABR can act as Ca(2+)o-dependent cofactors to enhance neuronal metabotropic glutamate signaling. PMID- 15550549 TI - Recurrent evolution of herbivory in small, cold-climate lizards: breaking the ecophysiological rules of reptilian herbivory. AB - Herbivory has evolved in many groups of vertebrates, but it is rare among both extinct and extant nonavian reptiles. Among squamate reptiles, (lizards, snakes, and their relatives), <2% of the >7,800 species are considered to be herbivorous, and herbivory is restricted to lizards. Here, we show that within a group of South American lizards (Liolaemidae, approximately 170 species), herbivory has evolved more frequently than in all other squamates combined and at a rate estimated to be >65 times faster. Furthermore, in contrast to other herbivorous lizards and to existing theory, most herbivorous liolaemids are small bodied and live in cool climates. Herbivory is generally thought to evolve only in reptile species that are large bodied, live in warm climates, and maintain high body temperatures. These three well known "rules" of herbivory are considered to form the bases of physiological constraints that explain the paucity of herbivorous reptile species. We suggest that the recurrent and paradoxical evolution of herbivory in liolaemids is explained by a combination of environmental conditions (promoting independent origins of herbivory in isolated cool-climate regions), ecophysiological constraints (requiring small body size in cool climates, yet high body temperatures for herbivores), and phylogenetic history. More generally, our study demonstrates how integrating information from ecophysiology and phylogeny can help to explain macroevolutionary trends. PMID- 15550550 TI - Mechanisms of spontaneous osteoclastogenesis in cancer with bone involvement. AB - Bone metastases represents a common cause of morbidity in patients suffering many types of cancer: breast, lung, kidney, prostate, and multiple myeloma. Osteolytic metastases often cause severe pain, pathologic fractures, hypercalcemia, spinal cord compression, and other nerve-compression syndromes. Osteoclasts (OCs), cells deriving from granulocitic-macrophagic lineage, are responsible for osteolysis, which may be reduced by inhibiting both OCs formation and activity. By studying bone osteolytic metastases mechanism in solid tumors, we report here our findings that cancer patients with bone involvement display an increase in osteoclasts precursors, compared with both healthy controls and cancer patients without bone metastases. Peripheral blood mononuclear cells (PBMCs) from patients with osteolytic lesions show osteoclastogenesis without adding M-CSF, RANKL, or TNF alpha. However, these factors are necessary to generate OCs from healthy donors, non-osteolytic patient PBMCs and T-cell depleted PBMCs. OCs derived from cancer patients show more resorption pits than OCs from healthy donors and express genes involved in osteoclastogenesis. Our data show that a spontaneous osteoclastogenesis occurs in patients affected by osteolytic lesions and may be supported by factors released by T lymphocytes. These factors could give a priming to osteoclast precursors and promote osteoclastogenesis. In fact, T-cell depleted PBMCs do not differentiate into OCs without adding M-CSF and RANKL. Moreover, we do not obtain a higher number of OCs by increasing RANKL doses in cultures, and OCs and T lymphocytes mRNA level are detected for TNF-alpha but not for RANKL. The addition of OPG to PBMCs cultures do not modify spontaneous osteoclastogenesis. A neutralizing anti-TNF-alpha antibody in unstimulated PBMC cultures of osteolytic cancer patients induces an inhibition of osteoclastogenesis. These data suggest that TNF-alpha may be responsible for osteoclastogenesis in these tumors. PMID- 15550551 TI - HO-1 induction restores c-AMP-dependent lung epithelial fluid transport following severe hemorrhage in rats. AB - Inhibition of cAMP-dependent stimulation of the vectorial fluid transport across the lung epithelium following hemorrhagic shock is mediated by NO released within the airspaces of the lung. We tested here the hypothesis that prior induction of HO-1 would attenuate the release of NO in the airspaces, thus preventing the inhibition of the c-AMP stimulation of alveolar fluid clearance (ALC) in rats. Indeed, HO-1 induction restored the cAMP-mediated up-regulation of ALC after hemorrhage by decreasing NO released within the airspaces of the lung. In vitro studies demonstrated that HO-1 induction significantly reduced the iNOS-mediated release of NO by alveolar macrophages stimulated with endotoxin for 24 h. This effect is explained in part by a HO-1-dependent attenuation of the LPS-mediated nuclear translocation of NF-kappaB. In addition, HO-1 induction also significantly reduced the iNOS-mediated release of NO by MH-S cells that were stimulated with interferon-gamma by decreasing the phosphorylation of STAT 1, another transcription factor important for the activation of the iNOS promoter. In contrast, HO-1 induction did not affect the production of NO by rat alveolar epithelial type II cells that were stimulated with cytomix (a mixture of TNF alpha, IL-1beta, and IFN-gamma) for 24 h. In summary, these results provide the first in vivo evidence that the induction of HO-1 in the lung restores a normal fluid transport capacity of the alveolar epithelium following hemorrhagic shock by inhibiting the iNOS-mediated release of NO by alveolar macrophages. PMID- 15550552 TI - Contribution of host MMP-2 and MMP-9 to promote tumor vascularization and invasion of malignant keratinocytes. AB - The matrix metalloproteinases (MMPs) play a key role in normal and pathological angiogenesis by mediating extracellular matrix degradation and/or controlling the biological activity of growth factors, chemokines, and/or cytokines. Specific functions of individual MMPs as anti- or proangiogenic mediators remain to be elucidated. In the present study, we assessed the impact of single or combined MMP deficiencies in in vivo and in vitro models of angiogenesis (malignant keratinocyte transplantation and the aortic ring assay, respectively). MMP-9 was predominantly expressed by neutrophils in tumor transplants, whereas MMP-2 and MMP-3 were stromal. Neither the single deficiency of MMP-2, MMP-3, or MMP-9, nor the combined absence of MMP-9 and MMP-3 did impair tumor invasion and vascularization in vivo. However, there was a striking cooperative effect in double MMP-2:MMP-9-deficient mice as demonstrated by the absence of tumor vascularization and invasion. In contrast, the combined lack of MMP-2 and MMP-9 did not impair the in vitro capillary outgrowth from aortic rings. These results point to the importance of a cross talk between several host cells for the in vivo tumor promoting and angiogenic effects of MMP-2 and MMP-9. Our data demonstrate for the first time in an experimental model that MMP-2 and MMP-9 cooperate in promoting the in vivo invasive and angiogenic phenotype of malignant keratinocytes. PMID- 15550553 TI - Cultured monolayers of the dog jejunum with the structural and functional properties resembling the normal epithelium. AB - The development of a culture of the normal mammalian jejunum motivated this work. Isolated crypt cells of the dog jejunum were induced to form primary cultures on Snapwell filters. Up to seven subcultures were studied under the electron microscope and in Ussing chambers. Epithelial markers were identified by RT-PCR, Western blot, and immunofluorescent staining. Confluent monolayers exhibit a dense apical brush border, basolateral membrane infoldings, desmosomes, and tight junctions expressing zonula occludens-1, occludin-1, and claudin-3 and -4. In OptiMEM medium fortified with epidermal growth factor, hydrocortisone, and insulin, monolayer transepithelial voltage was -6.8 mV (apical side), transepithelial resistance was 1,050 Omega.cm(2), and short-circuit current (I(sc)) was 8.1 microA/cm(2). Transcellular and paracellular resistances were estimated as 14.8 and 1.1 kOmega.cm(2), respectively. Serosal ouabain reduced voltage and current toward zero, as did apical amiloride. The presence of mRNA of alpha-epithelial Na(+) channel (ENaC) was confirmed. Na-d-glucose cotransport was identified with an antibody to Na(+)-glucose cotransporter (SGLT) 1. The unidirectional mucosa-to-serosa Na(+) flux (19 nmol.min(-1).cm(-2)) was two times as large as the reverse flux, and net transepithelial Na(+) flux was nearly double the amiloride-sensitive I(sc). In plain Ringer solution, the amiloride sensitive I(sc) went toward zero. Under these conditions plus mucosal amiloride, serosal dibutyryl-cAMP elicited a Cl(-)-dependent I(sc) consistent with the stimulation of transepithelial Cl(-) secretion. In conclusion, primary cultures and subcultures of the normal mammalian jejunum form polarized epithelial monolayers with 1) the properties of a leaky epithelium, 2) claudins specific to the jejunal tight junction, 3) transepithelial Na(+) absorption mediated in part by SGLT1 and ENaC, and 4) electrogenic Cl(-) secretion activated by cAMP. PMID- 15550548 TI - Coping with cold: An integrative, multitissue analysis of the transcriptome of a poikilothermic vertebrate. AB - How do organisms respond adaptively to environmental stress? Although some gene specific responses have been explored, others remain to be identified, and there is a very poor understanding of the system-wide integration of response, particularly in complex, multitissue animals. Here, we adopt a transcript screening approach to explore the mechanisms underpinning a major, whole-body phenotypic transition in a vertebrate animal that naturally experiences extreme environmental stress. Carp were exposed to increasing levels of cold, and responses across seven tissues were assessed by using a microarray composed of 13,440 cDNA probes. A large set of unique cDNAs (approximately 3,400) were affected by cold. These cDNAs included an expression signature common to all tissues of 252 up-regulated genes involved in RNA processing, translation initiation, mitochondrial metabolism, proteasomal function, and modification of higher-order structures of lipid membranes and chromosomes. Also identified were large numbers of transcripts with highly tissue-specific patterns of regulation. By unbiased profiling of gene ontologies, we have identified the distinctive functional features of each tissue's response and integrate them into a comprehensive view of the whole-body transition from one strongly adaptive phenotype to another. This approach revealed an expression signature suggestive of atrophy in cooled skeletal muscle. This environmental genomics approach by using a well studied but nongenomic species has identified a range of candidate genes endowing thermotolerance and reveals a previously unrecognized scale and complexity of responses that impacts at the level of cellular and tissue function. PMID- 15550554 TI - Ameliorating effect of hepatocyte growth factor on inflammatory bowel disease in a murine model. AB - Hepatocyte growth factor (HGF), a multifunctional cytokine, accelerates intestinal epithelial proliferation. We studied the effects of HGF in mice with trinitrobenzene sulfonic acid-induced colitis, which shows clinical and molecular resemblance to Crohn's disease. Mice with colitis repeatedly were transfected intramuscularly with human HGF cDNA. Weight, survival, histopathology, proinflammatory cytokine mRNAs, and leukocyte infiltration were assessed. Treatment with HGF cDNA induced tyrosine phosphorylation of intestinal c-Met/HGF receptors, inhibited apoptosis, and promoted mitosis in intestinal epithelial cells, accelerating intestinal epithelial restoration and suppressing inflammation. Transfection with HGF cDNA markedly suppressed intestinal mRNA expression of T-helper 1 cytokines such as interleukin-12 and -1beta, interferon gamma, and tumor necrosis factor-alpha. Numbers of total and CD4-positive T cells, neutrophils, and myloperoxidase activity in intestinal epithelium were diminished by HGF gene transfer, which also prevented weight loss, and improved survival. HGF might prove useful for controlling inflammatory bowel disease. PMID- 15550555 TI - Gender differences in small intestinal perfusion following trauma hemorrhage: the role of endothelin-1. AB - Although gender differences in intestinal perfusion exist following trauma hemorrhage (T-H), it remains unknown whether endothelin-1 (ET-1) plays any role in these dimorphic responses. To study this, male, proestrus female (female), and 17 beta-estradiol (E2)-treated male rats underwent midline laparotomy, hemorrhagic shock (blood pressure 40 mmHg, 90 min), and resuscitation (Ringer lactate, 4X shed blood volume, 1 h). Two hours thereafter, intestinal perfusion flow (IPF) was measured using isolated intestinal perfusion. The IPF in sham operated males was significantly lower than those in other groups and decreased markedly following T-H. In contrast, no significant decrease in IPF was observed in females and E2 males following T-H. The lower IPF in sham-operated males was significantly elevated by ET(A) receptor antagonist (BQ-123) administration and was similar to that seen in sham-operated females. The decreased IPF in males after T-H was also attenuated by BQ-123 administration. The intestinal ET-1 levels in sham-operated males were significantly higher than in other groups. Although plasma and intestinal ET-1 levels increased significantly after T-H in all groups, they were highest in males. Plasma E2 levels in females and E2 males were significantly higher than in males; however, they were not affected by T-H. There was a negative correlation between plasma ET-1 and E2 following T-H. Thus ET-1 appears to play an important role in intestinal perfusion failure following T-H in males. Because E2 can modulate this vasoconstrictor effect of ET-1, these findings may partially explain the previously observed salutary effect of estrogen in improving intestinal perfusion following T-H in males. PMID- 15550556 TI - Water and enzyme secretion are tightly coupled in pancreatic secretion stimulated by food or CCK-58 but not by CCK-8. AB - Pancreatic secretion of protein, water, chloride, and bicarbonate under basal conditions and in response to intravenous and intraduodenal stimuli were studied in awake rats fully recovered from surgery. During the basal phase of pancreatic secretion, protein output and water output were weakly correlated or uncorrelated, consistent with separate regulation and distinct cellular origin of enzyme (acinar cells) and water (duct cells), referred to as the two-component paradigm of pancreatic secretion. When pancreatic secretion was stimulated physiologically, water and protein output abruptly became strongly and significantly correlated, suggesting that protein secretion and water secretion are tightly coupled or that protein secretion is dependent on water secretion. The apparent function of this coupling is to resist or prevent increases in protein concentration as protein output increases. This pattern of secretion was reproduced by intravenous infusion of the CCK-58 form of cholecystokinin, which strongly stimulates pancreatic water and chloride secretion, but not by CCK-8, which only weakly stimulates water and chloride secretion in a non-dose-dependent manner. The remarkably tight association of water and protein secretion in food stimulated and CCK-58-stimulated pancreatic secretion is consistent with a single cell type as the origin of both water and enzyme secretion, i.e., the acinar cell, and is not consistent with the two-component paradigm of pancreatic secretion. Because CCK-58 is the only detectable endocrine form of cholecystokinin in the rat and its bioactivity pattern is markedly and qualitatively different from CCK-8, actions previously recorded for CCK-8 should be reexamined. PMID- 15550557 TI - Molecular determinants of the prothrombogenic phenotype assumed by inflamed colonic venules. AB - Although platelets have been implicated in the pathogenesis of human inflammatory bowel diseases, little is known about the magnitude of platelet accumulation in the inflamed bowel, what regulates this process, and its relevance to the overall inflammatory response. In this study, intravital video microscopy was used to monitor the trafficking of platelets and leukocytes and vascular permeability in colonic venules during the development of colonic inflammation induced by 3% dextran sodium sulfate (DSS). Blocking antibodies directed against different adhesion molecules as well as P-selectin-deficient mice were used to define the adhesive determinants of DSS-induced platelet recruitment. DSS induced an accumulation of adherent platelets that was temporally correlated with the appearance of adherent leukocytes and with disease severity. Platelet adhesion and, to a lesser extent, leukocyte adhesion were attenuated by immunoblockade of P-selectin and its ligand P-selectin glycoprotein ligand-1 (PSGL-1), with contributions from both platelet- and endothelial cell-associated P-selectin. DSS induced a rapid and sustained increase in vascular permeability that was greatly attenuated in P-selectin-deficient mice. P-selectin bone marrow chimeras revealed that both endothelial cell- and platelet-associated P-selectin contribute to the P-selectin expression detected in the inflamed colonic microvasculature, with endothelial P-selectin making a larger contribution. Our findings indicate that colonic inflammation is associated with the induction of a prothrombogenic phenotype in the colonic microcirculation, with P-selectin and its ligand PSGL-1 playing a major role in the recruitment of platelets. PMID- 15550558 TI - Inflammation induced changes in arachidonic acid metabolism in cat LES circular muscle. AB - Myogenic lower esophageal sphincter (LES) tone is maintained by arachidonic acid metabolites, such as PGF(2alpha) and thromboxane A(2)/B(2). Experimental esophagitis in cat reduces LES in vivo pressure and in vitro tone. Because IL 1beta may mediate esophagitis-associated reduction in ACh release in esophagus, we examined whether IL-1beta may also play a role in esophagitis-induced reduction of LES tone. A cat model of experimental esophagitis was obtained by repeated esophageal perfusion with HCl (Biancani P, Barwick K, Selling J, and McCallum R. Gastreonterology 87: 8-16, 1984 and Sohn UD, Harnett KM, Cao W, Rich H, Kim N, Behar J, and Biancani P. J Pharmacol Exp Ther 283: 1293-1304, 1997.). LES circular muscle strips were examined in muscle chambers as previously described (Biancani P, Billett G, Hillemeier C, Nissenshon M, Rhim BY, Sweczack S, and Behar J. Gastroenterology 103: 1199-1206, 1992). Levels of inflammatory mediators were measured. IL-1beta levels were higher in esophagitis than in normal LES. IL-1beta reduced normal LES tone, and the reduction was reversed by catalase, suggesting a role of H(2)O(2). This was confirmed by IL-1beta-induced production of H(2)O(2) in normal LES and elevated H(2)O(2) levels in esophagitis. H(2)O(2) by itself is sufficient to explain the changes that occur in the muscle, reducing its ability to contract. H(2)O(2) increased PGE(2) in normal LES, and PGE(2) levels were elevated in esophagitis LES, whereas PGF(2alpha) levels were unchanged. H(2)O(2) also increased levels of 8-isoprostanes, stable prostaglandin like compounds formed by free radical-induced peroxidation of arachidonic acid, and 8-isoprostane levels were elevated in esophagitis. The PGF(2alpha) analog 8 iso-PGF(2alpha) caused little contraction of LES strips but reduced PGF(2alpha) binding and contraction of normal LES. In esophagitis, PGF(2alpha) binding and contraction were reduced in LES, suggesting that isoprostanes may contribute to reduction in tone in esophagitis. The data suggest that, in esophagitis, IL-1beta causes production of H(2)O(2). H(2)O(2) increases PGE(2), which relaxes the LES, and 8-iso-F(2alpha), which blocks PGF(2alpha)-mediated contraction. PMID- 15550559 TI - Sinusoidal endothelial COX-1-derived prostanoids modulate the hepatic vascular tone of cirrhotic rat livers. AB - CCl(4) cirrhotic rat liver exhibits a hyperresponse to the alpha(1)-adrenergic agonist methoxamine (Mtx) that is associated with enhanced thromboxane A(2) (TXA(2)) production and is abrogated by indomethacin. To further elucidate the molecular mechanisms involved in the hyperresponse to vasoconstrictors, portal perfusion pressure dose-response curves to Mtx were performed in CCl(4) cirrhotic rats livers after preincubation with vehicle, the cyclooxygenase (COX)-1 selective inhibitor SC-560, and the COX-2 selective inhibitor SC-236. TXA(2) production was determined in samples of the perfusate. COX-1 expression was analyzed and quantified in hepatocytes, Kupffer cells, sinusoidal endothelial cells (SEC), and hepatic stellate cells (HSC) isolated from control and cirrhotic rat livers by double-immunofluorescence staining, with specific markers for each population using flow cytometry or Western blot analysis. COX-1 protein levels were not significantly increased in cirrhotic livers, but COX-2 protein expression was increased. COX-1 inhibition, but not COX-2, significantly attenuated the response to Mtx and prevented the increased production of TXA(2). Cirrhotic livers showed an increased expression of COX-1 in SEC and reduced expression in HSC compared with control livers, whereas COX-1 was similarly distributed in Kupffer cells. Despite abundant hepatic COX-2 expression, the increased response to Mtx of cirrhotic livers is mainly dependent of COX-1. Upregulation of COX-1 in cirrhotic SEC may be responsible for the hyperesponse to Mtx. PMID- 15550560 TI - IL-7 exacerbates chronic colitis with expansion of memory IL-7Rhigh CD4+ mucosal T cells in mice. AB - We have previously demonstrated that mucosal CD4(+) T cells expressing high levels of IL-7 receptor (IL-7R(high)) are pathogenic cells responsible for chronic colitis. Here we investigate whether IL-7 is directly involved in the expansion of IL-7R(high) memory CD4(+) mucosal T cells and the exacerbation of colitis. We first showed that CD4(+) lamina propria lymphocytes (LPLs) from wild type, T cell receptor-alpha-deficient (TCR-alpha(-/-)), and recombinase activating gene (RAG)-2(-/-)-transferred mice with or without colitis showed phenotypes of memory cells, but only CD4(+) LPLs from colitic mice showed IL 7R(high). In vitro stimulation by IL-7, but not by IL-15 and thymic stromal lymphopoietin, enhanced significant proliferative responses and survival of colitic CD4(+), but not normal CD4(+) LPLs. Importantly, in vivo administration of IL-7 mice accelerated the expansion of IL-7R(high) memory CD4(+) LPLs and thereby exacerbated chronic colitis in RAG-2(-/-) mice transferred with CD4(+) LPLs from colitic TCR-alpha(-/-) mice. Conversely, the administration of anti-IL 7R monoclonal antibody significantly inhibited the development of TCR-alpha(-/-) colitis with decreased expansion of CD4(+) LPLs. Collectively, the present data indicate that IL-7 is essential for the expansion of pathogenic memory CD4(+) T cells under pathological conditions. Therefore, therapeutic approaches targeting the IL-7R pathway may be feasible in the treatment of human inflammatory bowel disease. PMID- 15550561 TI - Basolateral Mg2+/Na+ exchange regulates apical nonselective cation channel in sheep rumen epithelium via cytosolic Mg2+. AB - High potassium diets lead to an inverse regulation of sodium and magnesium absorption in ruminants, suggesting some form of cross talk. Previous Ussing chamber experiments have demonstrated a divalent sensitive Na(+) conductance in the apical membrane of ruminal epithelium. Using patch-clamped ruminal epithelial cells, we could observe a divalent sensitive, nonselective cation conductance (NSCC) with K(+) permeability > Cs(+) permeability > Na(+) permeability. Conductance increased and rectification decreased when either Mg(2+) or both Ca(2+) and Mg(2+) were removed from the internal or external solution or both. The conductance could be blocked by Ba(2+), but not by tetraethylammonium (TEA). Subsequently, we studied this conductance measured as short-circuit current (I(sc)) in Ussing chambers. Forskolin, IBMX, and theophylline are known to block both I(sc) and Na transport across ruminal epithelium in the presence of divalent cations. When the NSCC was stimulated by removing mucosal calcium, an initial decrease in I(sc) was followed by a subsequent increase. The cAMP-mediated increase in I(sc) was reduced by low serosal Na(+) and serosal addition of imipramine or serosal amiloride and depended on the availability of mucosal magnesium. Luminal amiloride had no effect. Flux studies showed that low serosal Na(+) reduced (28)Mg fluxes from mucosal to serosal. The data suggest that cAMP stimulates basolateral Na(+)/Mg(2+) exchange, reducing cytosolic Mg. This increases sodium uptake through a magnesium-sensitive NSCC in the apical membrane. Likewise, the reduction in magnesium uptake that follows ingestion of high potassium fodder may facilitate sodium absorption, as observed in studies of ruminal osmoregulation. Possibly, grass tetany (hypomagnesemia) is a side effect of this useful mechanism. PMID- 15550562 TI - Components of intestinal epithelial hypoxia activate the virulence circuitry of Pseudomonas. AB - We have previously shown that a lethal virulence trait in Pseudomonas aeruginosa, the PA-I lectin, is expressed by bacteria within the intestinal lumen of surgically stressed mice. The aim of this study was to determine whether intestinal epithelial hypoxia, a common response to surgical stress, could activate PA-I expression. A fusion construct was generated to express green fluorescent protein downstream of the PA-I gene, serving as a stable reporter strain for PA-I expression in P. aeruginosa. Polarized Caco-2 monolayers were exposed to ambient hypoxia (0.1-0.3% O2) for 1 h, with or without a recovery period of normoxia (21% O2) for 2 h, and then inoculated with P. aeruginosa containing the PA-I reporter construct. Hypoxic Caco-2 monolayers caused a significant increase in PA-I promoter activity relative to normoxic monolayers (165% at 1 h; P < 0.001). Similar activation of PA-I was also induced by cell free apical, but not basal, media from hypoxic Caco-2 monolayers. PA-I promoter activation was preferentially enhanced in bacterial cells that physically interacted with hypoxic epithelia. We conclude that the virulence circuitry of P. aeruginosa is activated by both soluble and contact-mediated elements of the intestinal epithelium during hypoxia and normoxic recovery. PMID- 15550563 TI - Cytokine regulation of human sterol 12alpha-hydroxylase (CYP8B1) gene. AB - Sterol 12alpha-hydroxylase (CYP8B1) catalyzes cholic acid synthesis in the liver and is feedback inhibited by bile acids. In addition to activating farnesoid X receptor (nuclear receptor subfamily 1H4), bile acids also induce inflammatory cytokines in hepatocytes. The objective of this study was to investigate the mechanism by which inflammatory cytokines inhibit human CYP8B1 gene transcription. Real-time PCR assays revealed that both chenodeoxycholic acid (CDCA) and interleukin-1beta (IL-1beta) markedly reduced CYP8B1, cholesterol 7alpha-hydroxylase CYP7A1 and hepatic nuclear factor 4alpha (HNF4alpha) mRNA expression levels in human primary hepatocytes. However, CDCA induced, but IL 1beta reduced, small heterodimer partner (SHP) mRNA expression. IL-1beta inhibited human CYP8B1 reporter activity only in liver cells, and a c-Jun NH(2) terminal kinase (JNK)-specific inhibitor-blocked IL-1beta inhibition. Activated JNK1 or c-Jun inhibited, whereas their dominant negative forms blocked, IL-1beta inhibition of CYP8B1 transcription. Mutagenesis analyses mapped an IL-1beta response element to a previously identified bile acid response element, which contains an HNF4alpha binding site. A dominant negative HNF4alpha inhibited CYP8B1 gene transcription and ectopically expressed HNF4alpha blocked IL-1beta inhibition. Furthermore, IL-1beta inhibited HNF4alpha gene transcription, protein expression, and binding to the CYP8B1 gene. JNK1 phosphorylated HNF4alpha and a JNK-specific inhibitor blocked the IL-1beta inhibition of HNF4alpha expression. These results suggest that IL-1beta inhibits CYP8B1 gene transcription via a mitogen-activated protein kinase/JNK pathway that inhibits HNF4alpha gene expression and its DNA-binding ability. This mechanism may play an important role in the adaptive response to inflammatory cytokines and in the protection of the liver during cholestasis. PMID- 15550564 TI - Surfactant proteins and inflammation: the yin and the yang. PMID- 15550565 TI - The stability of Seeman JX DNA topoisomers of paranemic crossover (PX) molecules as a function of crossover number. AB - We use molecular dynamics simulations in explicit water and salt (Na+) to determine the effect of varying the number of crossover points on the structure and stability of the PX65 paranemic crossover DNA molecule and its JXM topoisomers (M denotes the number of missing crossover points), recently synthesized by the Seeman group at New York University. We find that PX65, with six crossover points, is the most stable, and that the stability decreases monotonically with the number of crossover points PX65 > JX1 > JX2 > JX3 > JX4, with 6, 5, 4, 3 and 2 crossover points, respectively. Thus, for PX65/JX1, the strain energy is approximately 3 kcal/mol/bp, while it is approximately 13 kcal/mol/bp for JX2, JX3 and JX4. Another measure of the stability is the change in the structure from the minimum energy structure to the equilibrium structure at 300 K, denoted as root-mean-square deviation in coordinates (CRMSD). We find that CRMSD is approximately 3.5 A for PX65, increases to 6 A for JX1 and increases to 10 A for JX2/JX3/JX4. As the number of crossover points decreases, the distance between the two double helical domains of the PX/JX molecules increases from approximately 20 A for PX65 to 23 A for JX4. This indicates that JX2, JX3 and JX4 are less likely to form, at least in with Na+. However, in all the cases, the two double helical domains have average helicoidal parameters similar to a typical B-DNA of similar length and base sequence. PMID- 15550566 TI - Regulation of 6S RNA biogenesis by switching utilization of both sigma factors and endoribonucleases. AB - In Escherichia coli, 6S RNA functions as a modulator of RNA polymerase sigma70 holoenzyme activity, but its biosynthetic pathway remains uncharacterized. In this study, to further understand the regulatory circuit of 6S RNA biosynthesis for the modulation of Esigma70 activity, we have characterized the biogenesis of 6S RNA. We reveal that there are two different precursors, a long and a short molecule, which are transcribed from the distal P2 and proximal P1 promoter, respectively. Transcription from the P2 promoter is both sigma70- and sigmaS dependent, whereas, in contrast, P1 transcription is sigma70- but not sigmaS dependent. Both precursors are processed to generate the 5' end of 6S RNA, and while the long precursor is processed exclusively by RNase E, the short precursor is processed by both RNase G and RNase E. Our data indicate that the switching of the utilization of both sigma factors and endoribonucleases in the biogenesis of 6S RNA would play an essential role in modulating its levels in E.coli. PMID- 15550567 TI - Solution structure of a dsDNA:LNA triplex. AB - We have determined the NMR structure of an intramolecular dsDNA:LNA triplex, where the LNA strand is composed of alternating LNA and DNA nucleotides. The LNA oligonucleotide binds to the dsDNA duplex in the major groove by formation of Hoogsteen hydrogen bonds to the purine strand of the duplex. The structure of the dsDNA duplex is changed to accommodate the LNA strand, and it adopts a geometry intermediate between A- and B-type. There is a substantial propeller twist between base-paired nucleobases. This propeller twist and a concomitant large propeller twist between the purine and LNA strands allows the pyrimidines of the LNA strand to interact with the 5'-flanking duplex pyrimidines. Altogether, the triplex has a regular global geometry as shown by a straight helix axis. This shows that even though the third strand is composed of alternating DNA and LNA monomers with different sugar puckers, it forms a seamless triplex. The thermostability of the triplex is increased by 19 degrees C relative to the unmodified DNA triplex at acidic pH. Using NMR spectroscopy, we show that the dsDNA:LNA triplex is stable at pH 8, and that the triplex structure is identical to the structure determined at pH 5.1. PMID- 15550568 TI - DNA recombination with a heterospecific Cre homolog identified from comparison of the pac-c1 regions of P1-related phages. AB - Sequencing of the 7 kb immC region from four P1-related phages identified a novel DNA recombinase that exhibits many Cre-like characteristics, including recombination in mammalian cells, but which has a distinctly different DNA specificity. DNA sequence comparison to the P1 immC region showed that all phages had related DNA terminase, C1 repressor and DNA recombinase genes. Although these genes from phages P7, phi(w39) and p15B were highly similar to those from P1, those of phage D6 showed significant divergence. Moreover, the D6 sequence showed evidence of DNA deletion and substitution in this region relative to the other phages. Characterization of the D6 site-specific DNA recombinase (Dre) showed that it was a tyrosine recombinase closely related to the P1 Cre recombinase, but that it had a distinct DNA specificity for a 32 bp DNA site (rox). Cre and Dre are heterospecific: Cre did not catalyze recombination at rox sites and Dre did not catalyze recombination at lox sites. Like Cre, Dre catalyzed both integrative and excisive recombination and required no other phage-encoded proteins for recombination. Dre-mediated recombination in mammalian cells showed that, like Cre, no host bacterial proteins are required for efficient Dre-mediated site specific DNA recombination. PMID- 15550569 TI - Functional role of G9a-induced histone methylation in small heterodimer partner mediated transcriptional repression. AB - Site-specific modification of nucleosomal histones plays a central role in the formation of transcriptionally active and inactive chromatin structures. These modifications may serve as specific recognition motifs for chromatin proteins, which act as a signal for the adoption of the appropriate regulatory responses. Here, we show that the orphan nuclear receptor SHP (small heterodimer partner), a coregulator that inhibits the activity of several nuclear receptors, can associate with unmodified and lysine 9-methylated histone-3, but not with the acetylated protein. The naturally occurring SHP mutant (R213C), which exhibits decreased transrepression potential, interacts less avidly with K9-methylated histone 3. We demonstrate that SHP can functionally interact with histone deacetylase-1 and the G9a methyltransferase and that it is localized exclusively in nuclease-sensitive euchromatin. The results point to the involvement of a multistep mechanism in SHP-dependent transcriptional repression, which includes histone deacetylation, followed by H3-K9 methylation and stable association of SHP itself with chromatin. PMID- 15550570 TI - Attenuation of behavioral effects of cocaine by the Metabotropic Glutamate Receptor 5 Antagonist 2-Methyl-6-(phenylethynyl)-pyridine in squirrel monkeys: comparison with dizocilpine. AB - Growing evidence suggests a role for metabotropic glutamate receptors (mGluRs) in the behavioral effects of cocaine related to its abuse. The mGluR5 subtype, in particular, has come under scrutiny due to its distribution in brain regions associated with drug addiction. This study investigated interactions between the selective mGluR5 antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP) and cocaine in squirrel monkeys whose lever-pressing behavior was 1) maintained under a second-order schedule of cocaine self-administration, 2) extinguished and then reinstated by cocaine priming, and 3) controlled by the discriminative stimulus (DS) effects of cocaine. Additional studies determined the effects of MPEP on unconditioned behaviors, coordination, and muscle resistance. In each experiment, the effects of MPEP were compared with those of the N-methyl-d-aspartate antagonist dizocilpine. MPEP attenuated cocaine self-administration, cocaine induced reinstatement of drug seeking, and the DS effects of cocaine at doses that did not markedly impair motor function or operant behavior in the context of drug discrimination. Dizocilpine also attenuated cocaine self-administration, but it did not significantly alter cocaine-induced reinstatement of drug seeking, and it enhanced rather than attenuated the DS effects of cocaine. The findings point to a significant contribution of mGluR5 mechanisms in the behavioral effects of cocaine related to its abuse and suggest that MPEP has properties of a functional cocaine antagonist, which are not secondary to antagonism at NMDA receptors. The contrasting interactions of MPEP and dizocilpine with cocaine imply that glutamate acting through different metabotropic and ionotropic receptors may modulate the behavioral effects of cocaine in qualitatively different ways. PMID- 15550571 TI - Raloxifene relaxes rat pulmonary arteries and veins: roles of gender, endothelium, and antagonism of Ca2+ influx. AB - Effects of raloxifene have been documented in the systemic circulation. However, its impact on the pulmonary circulation is unclear. The present study investigated the role of gender, endothelial modulation, and Ca(2+) channel in relaxations evoked by raloxifene in rat pulmonary arteries and veins. Vascular responses were studied on isolated pulmonary blood vessels mounted in a myograph and constricted by U46619 (9,11-dideoxy-11alpha,9alpha-epoxymethanoprostaglandin F(2alpha)). Constrictions to CaCl(2) were studied in Ca(2+)-free, 60 mM K(+) solution. Changes in the intracellular calcium ion concentration ([Ca(2+)](i)) in vascular smooth muscle were measured using a calcium fluorescence imaging method. Raloxifene was more effective in relaxing U46619-constricted pulmonary arteries from male than female rats. Raloxifene-induced relaxation was unaffected by ICI 182,780 [7alpha-[9-[(4,4,5,5,5,-pentafluoropentyl)-sulfinyl]nonyl]-estra 1,3,5(10)-triene-3,17beta-diol], inhibition of the nitric oxide (NO) pathway, or removal of the endothelium. In arteries without endothelium, raloxifene attenuated CaCl(2)-induced constriction and CaCl(2)-stimulated increase in [Ca(2+)](i) with similar potencies. Raloxifene caused endothelium-independent relaxations in pulmonary veins, albeit to a lesser degree than in pulmonary arteries. The venous responses showed a gender difference because raloxifene was more potent in male veins. In summary, raloxifene relaxed rat pulmonary arteries, and this effect did not involve the endothelium/NO or ICI 182,780-sensitive estrogen receptors. Raloxifene, like nifedipine, reduced constriction and [Ca(2+)](i) increase in response to CaCl(2) in high K(+) solution. Raloxifene also relaxed high K(+)-constricted pulmonary veins. Our data indicate that raloxifene acutely relaxes rat pulmonary blood vessels primarily via inhibition of Ca(2+) influx through voltage-sensitive Ca(2+) channels. Finally, raloxifene induced more relaxation in blood vessels isolated from male than female rats. PMID- 15550572 TI - Self-Administration of cocaine-opioid combinations by rhesus monkeys: evaluation of the role of mu receptor efficacy using labor supply analysis. AB - Cocaine and heroin often are abused by self-administering the drugs in combination as a "speedball". We evaluated the extent to which intrinsic efficacy at the mu-opioid receptor influences combined cocaine-opioid self-administration and used the behavioral economic model termed "labor supply" to quantitatively evaluate the reinforcing effects of cocaine-opioid combinations. Rhesus monkeys (n = 8) were trained under a progressive-ratio schedule of i.v. cocaine injection in which the response requirement increased during the experimental session and the initial response requirement was varied. Combination of cocaine with heroin enhanced self-administration compared with the drugs individually, with ineffective doses of both drugs maintaining self-administration when combined. These effects also were observed with the high-efficacy mu agonist alfentanil and low-efficacy agonist nalbuphine. Using the labor supply economic model, combinations of heroin, alfentanil, or nalbuphine with relatively low doses of cocaine were found to increase the number of injections per session ("income") and total responses per session ("labor"). Combination of a relatively high dose of cocaine with either heroin or alfentanil, but not nalbuphine, also resulted in only a small reduction in income concomitant with increased labor, suggesting that heroin and alfentanil made cocaine consumption more resistant to increasing response costs, or more "inelastic." Collectively, these findings suggest that speedball self-administration may occur even with relatively low levels of intrinsic efficacy at mu-opioid receptors and that an inelastic relationship between drug consumption and labor may contribute to the persistence of speedball abuse. PMID- 15550573 TI - Postnatal inorganic lead exposure reduces midbrain dopaminergic impulse flow and decreases dopamine D1 receptor sensitivity in nucleus accumbens neurons. AB - Lead treatment via drinking water for 3 to 6 weeks at 250 ppm was found to significantly decrease the number of spontaneously active dopamine (DA) neurons in both the substantia nigra and ventral tegmental area that were recorded using standard extracellular electrophysiological recording techniques. Lead exposure did not affect the discharge rate or discharge pattern of these DA neurons. No significant decrease in the number of tyrosine hydroxylase immunopositive cells was detected in lead-treated animals relative to controls even though the length of lead exposure was extended beyond that of the electrophysiological studies. The significant lead-induced decrease in spontaneously active cells observed in the electrophysiological studies was, therefore, not due to cell death. An acute drug challenge with the DA receptor agonist apomorphine at a dose known to hyperpolarize midbrain DA neurons (50 mug/kg i.v.) was used to determine whether hyperpolarization would normalize the number of spontaneously active DA neurons. The results suggest that depolarization inactivation was most likely not the cause for this lead effect. The D(1) receptor agonist SKF-38393 [1-phenyl-2,3,4,5 tetrahydro-(1H)-3-benzazepine-7,8-diol] was iontophoretically applied to type I nucleus accumbens (Nacb) neurons. The results demonstrated that type I Nacb neurons have a significantly lower basal discharge rate in lead-treated animals relative to controls and that the Nacb DA D(1) receptors were significantly less sensitive to SKF-38393 in the lead-treated animals. Therefore, lead exposure decreases DA neuron impulse flow presynaptically and decreases DA D(1) receptor sensitivity postsynaptically in the nucleus accumbens. PMID- 15550574 TI - Pharmacological effects of ATI22-107 [2-(2-{2-[2-chloro-4-(6-oxo-1,4,5,6 tetrahydro-pyridazin-3-yl)-phenoxy]-acetylamino}-ethoxymethyl)-4-(2-chloro phenyl)-6-methyl-1,4-dihydro-pyridine-3,5-dicarboxylic acid dimethyl ester)], a novel dual pharmacophore, on myocyte calcium cycling and contractility. AB - Historically, inhibitors of type III phosphodiesterases (PDE-III) have been effective inotropes in mammalian myocardium, but their clinical utility has been limited by adverse events, including arrhythmias that are considered to be due to Ca(2+) overload. ATI22-107 [2-(2-{2-[2-chloro-4-(6-oxo-1,4,5,6-tetrahydro pyridazin-3-yl)-phenoxy]-acetylamino}-ethoxymethyl)-4-(2-chlorophenyl)-6-methyl 1,4-dihydro-pyridine-3,5-dicarboxylic acid dimethyl ester)], a novel, dual pharmacophore compound, was designed to simultaneously inhibit the cardiac phosphodiesterase (PDE-III) and produce inotropic effects, whereas inhibiting the L-type calcium channel (LTCC) was designed to minimize increases in diastolic Ca(2+). We compared the effects of ATI22-107 and enoximone, a pure PDE-III inhibitor, on the Fluo-3 calcium transient in normal feline ventricular myocytes and trabeculae. Enoximone-induced dose-dependent increases in peak [Ca(2+)](i), diastolic [Ca(2+)](i), T50, and T75. ATI22-107 demonstrated similar dose dependent increases in peak [Ca(2+)](i) at 300 nM and 1.0 microM doses, with no further increases at higher doses. Throughout the dosing range, ATI22-107 induced much smaller, if any, increases in diastolic [Ca(2+)](i), T(25), and T(75). Current measurement of LTCC via patch-clamp techniques revealed dose-dependent decreases in LTCC current with an increasing dose of ATI22-107, thereby validating the dual functionality of the drug that has been proposed in this study. Studies in isolated trabeculae demonstrated that enoximone-induced a dose dependent augmentation of the entire force-frequency relation in normal myocardium, whereas augmentation of contractility was only observed at lower stimulation frequencies with ATI22-107. These results demonstrate the effects of the LTCC-antagonizing moiety of ATI22-107 and suggest that the novel simultaneous combination of PDE-III and LTCC inhibition by one molecule may produce a favorable profile of limited inotropy without detrimental effects of increased diastolic [Ca(2+)](i). PMID- 15550575 TI - First demonstration of a functional role for central nervous system betaine/{gamma}-aminobutyric acid transporter (mGAT2) based on synergistic anticonvulsant action among inhibitors of mGAT1 and mGAT2. AB - In a recent study, EF1502 [N-[4,4-bis(3-methyl-2-thienyl)-3-butenyl]-3-hydroxy-4 (methylamino)-4,5,6,7-tetrahydrobenzo [d]isoxazol-3-ol], which is an N substituted analog of the GAT1-selective GABA uptake inhibitor exo-THPO (4-amino 4,5,6,7-tetrahydrobenzo[d]isoxazol-3-ol), was found to inhibit GABA transport mediated by both GAT1 and GAT2 in human embryonic kidney (HEK) cells expressing the mouse GABA transporters GAT1 to 4 (mGAT1-4). In the present study, EF1502 was found to possess a broad-spectrum anticonvulsant profile in animal models of generalized and partial epilepsy. When EF1502 was tested in combination with the clinically effective GAT1-selective inhibitor tiagabine [(R)-N-[4,4-bis(3-methyl 2-thienyl)-3-butenyl]nipecotic acid] or LU-32-176B [N-[4,4-bis(4-fluorophenyl) butyl]-3-hydroxy-4-amino-4,5,6,7-tetrahydrobenzo[d]isoxazol-3-ol], another GAT1 selective N-substituted analog of exo-THPO, a synergistic rather than additive anticonvulsant interaction was observed in the Frings audiogenic seizure susceptible mouse and the pentylenetetrazol seizure threshold test. In contrast, combination of the two mGAT1-selective inhibitors, tiagabine and LU-32-176B, resulted in only an additive anticonvulsant effect. Importantly, the combination of EF1502 and tiagabine did not result in a greater than additive effect in the rotarod behavioral impairment test. In subsequent in vitro studies conducted in HEK-293 cells expressing the cloned mouse GAT transporters mGAT1 and mGAT2, EF1502 was found to noncompetitively inhibit both mGAT1 and the betaine/GABA transporter mGAT2 (K(i) of 4 and 5 muM, respectively). Furthermore, in a GABA release study conducted in neocortical neurons, EF1502 did not act as a substrate for the GABA carrier. Collectively, these findings support a functional role for mGAT2 in the control of neuronal excitability and suggest a possible utility for mGAT2-selective inhibitors in the treatment of epilepsy. PMID- 15550576 TI - Signal transduction blockade and cancer: combination therapy or multi-targeted inhibitors? PMID- 15550577 TI - Clinical trial design for microarray predictive marker discovery and assessment. AB - Transcriptional profiling technologies that simultaneously measure the expression of thousands of mRNA species represent a powerful new clinical research tool. Similar to previous laboratory analytical methods including immunohistochemistry, PCR and in situ hybridization, this new technology may also find its niche in routine diagnostics. Outcome predictors discovered by these methods may be quite different from previous single-gene markers. These novel tests will probably combine the information embedded in the expression of multiple genes with mathematical prediction algorithms to formulate classification rules and predict outcome. The performance of machine learning-algorithm-based diagnostic tests may improve as they are trained on larger and larger sets of samples, and several generations of tests with improving accuracy may be introduced sequentially. Several gene-expression profiling-technology platforms are mature enough for clinical testing. The most important next step that is needed for further progress is the development and validation of multigene predictors in prospectively designed clinical trials to determine the true accuracy and clinical value of this new technology. This manuscript reviews methodological and statistical issues relevant to clinical trial design to discover and validate multigene predictors of response to therapy. PMID- 15550578 TI - Recent advances in endocrine therapy for postmenopausal women with early breast cancer: implications for treatment and prevention. AB - BACKGROUND: In the treatment of advanced breast cancer, third-generation aromatase inhibitors (AIs) have shown superior efficacy and tolerability compared with tamoxifen and megestrol acetate, the previous standard endocrine therapies in the first- and second-line settings, respectively. AIs are now being assessed in the adjuvant and prevention settings. DESIGN: Literature review (PubMed search). RESULTS: Tamoxifen is currently the only endocrine option available for adjuvant therapy and chemoprevention in postmenopausal women. However, results from the ATAC ('Arimidex', Tamoxifen, Alone or in Combination) trial have shown anastrozole to be more effective than tamoxifen as adjuvant therapy for postmenopausal women with hormone-responsive early breast cancer. Other third generation AIs, including letrozole and exemestane, are also being investigated as adjuvant therapies. In the chemoprevention setting, tamoxifen is the only available endocrine option for women at high risk of breast cancer but, given that these are healthy subjects, is associated with an unacceptable rate of adverse events. Raloxifene is being further assessed in the STAR (Study of Tamoxifen and Raloxifene) trial, while anastrozole is being evaluated in the second IBIS-II (International Breast Intervention Study II). CONCLUSIONS: AIs, in particular anastrozole, are set to change the way that early breast cancer is treated. Effective and better-tolerated endocrine alternatives for breast cancer prevention may become available in the future. PMID- 15550579 TI - Toremifene and tamoxifen are equally effective for early-stage breast cancer: first results of International Breast Cancer Study Group Trials 12-93 and 14-93. AB - BACKGROUND: Toremifene is a chlorinated derivative of tamoxifen, developed to improve its risk-benefit profile. The International Breast Cancer Study Group (IBCSG) conducted two complementary randomized trials for peri- and postmenopausal patients with node-positive breast cancer to compare toremifene versus tamoxifen as the endocrine agent and simultaneously investigate a chemotherapy-oriented question. This is the first report of the endocrine comparison after a median follow-up of 5.5 years. PATIENTS AND METHODS: 1035 patients were available for analysis: 75% had estrogen receptor (ER)-positive primary tumors, the median number of involved axillary lymph nodes was three and 81% received prior adjuvant chemotherapy. RESULTS: Toremifene and tamoxifen yielded similar disease-free (DFS) and overall survival (OS): 5-year DFS rates of 72% and 69%, respectively [risk ratio (RR)=0.95; 95% confidence interval (CI)=0.76-1.18]; 5-year OS rates of 85% and 81%, respectively (RR = 1.03; 95% CI = 0.78-1.36). Similar outcomes were observed in the ER-positive cohort. Toxicities were similar in the two treatment groups with very few women (<1%) experiencing severe thromboembolic or cerebrovascular complications. Quality of life results were also similar. Nine patients developed early stage endometrial cancer (toremifene, six; tamoxifen, three). CONCLUSIONS: Toremifene is a valid and safe alternative to tamoxifen in postmenopausal women with endocrine responsive breast cancer. PMID- 15550580 TI - Capecitabine and vinorelbine in elderly patients (> or =65 years) with metastatic breast cancer: a phase I trial (SAKK 25/99). AB - BACKGROUND: Few chemotherapy regimens are suitable for the treatment of elderly patients with advanced breast cancer. With the aim of finding a regimen with a low burden of subjective non-overlapping toxic effects, vinorelbine and capecitabine were chosen to be investigated in a phase I dose-finding study. PATIENTS AND METHODS: Thirty-six patients with advanced breast cancer were stratified for the presence of bone and non-bone involvement and treated at four dose levels from capecitabine 800 mg/m2 orally days 1-14 and vinorelbine 20 mg/m2 intravenously days 1 and 8, to capecitabine 1250 mg/m2 orally days 1-14 and vinorelbine 25 mg/m2 intravenously days 1 and 8, for a maximum of six cycles. None of the patients had received prior chemotherapy for metastatic/advanced disease. Fifty-three per cent of patients with bone metastases and 67% of patients without bone metastases had visceral disease. The median age was 70 years for the 15 with bone involvement patients and 73 years for the 21 without bone involvement patients. RESULTS: Twenty-eight patients were fully evaluable for hematological dose-limiting toxicity (DLT), and all patients for other DLTs and for antitumor activity. One DLT with grade 3 venous thrombosis at dose level 2 and two dose-limiting neutropenia events at level 3 occurred in patients without bone involvement. Two dose-limiting neutropenia events were observed at dose level 2 for patients with bone involvement. Thus, the recommended dose was defined at level 1 (capecitabine 1000 mg/m2 days 1-14 and vinorelbine 20 mg/m2 days 1 and 8) for patients with bone involvement. For patients without bone involvement, the recommended dose was at level 2 (capecitabine 1250 mg/m2 days 1 14 and vinorelbine 20 mg/m2 days 1 and 8). For patients without bone involvement the overall response rate was 48% and the time to progression (TTP) was 4.5 months [95% confidence interval (CI) 3.3-6.9]. For patients with bone involvement the overall response rate was 53% and TTP was 5.3 months (95% CI 2.7-7.8). CONCLUSIONS: This regimen of capecitabine and vinorelbine is well tolerated and effective in elderly patients with metastatic breast cancer. Toxicity was mainly hematological and was observed at a lower dose in patients with bone involvement. A phase II study with the two different dose levels for elderly patients with and without bone involvement is currently being conducted. PMID- 15550581 TI - First-line treatment of metastatic colorectal cancer with irinotecan, oxaliplatin and 5-fluorouracil/leucovorin (FOLFOXIRI): results of a phase II study with a simplified biweekly schedule. AB - BACKGROUND: In a previous phase I-II study we demonstrated that the FOLFOXIRI regimen [irinotecan 125-175 mg/m2 day 1, oxaliplatin 100 mg/m2 day 1, l leucovorin (l-LV) 200 mg/m2 day 1, 5-fluorouracil (5-FU) 3800 mg/m2 as a 48-h chronomodulated continuous infusion starting on day 1, repeated every 2 weeks] has promising activity and efficacy in metastatic colorectal cancer. However, this regimen required a chronomodulated infusion of 5-FU, and because neutropenia occurred in 32% of cycles, granulocyte colony-stimulating factor (G-CSF) was used and the delivered dose intensity was only approximately 78% of planned. Therefore, we conducted the present phase II study in order to develop a simplified FOLFOXIRI regimen that could be more easily administered in clinical practice as well as in multicenter settings. PATIENTS AND METHODS: A total of 32 patients with unresectable metastatic colorectal cancer received irinotecan 165 mg/m2 day 1, oxaliplatin 85 mg/m2 day 1, l-LV 200 mg/m2 day 1 and 5-FU 3200 mg/m2 as a 48-h continuous (not chronomodulated) infusion starting on day 1, repeated every 2 weeks. RESULTS: All 32 patients were evaluated for safety and the incidence of grade 3-4 toxic effects, and the use of G-CSF seemed to be lower than with the previous FOLFOXIRI regimen: grade 4 neutropenia (34%), grade 3 diarrhea (16%), grade 3 stomatitis (6%) and grade 2-3 peripheral neurotoxicity (37%) were reported, and G-CSF was used in 23% of cycles. Delivered dose intensity was 88% of that planned, and no toxic deaths occurred. The intention-to treat analysis for activity showed four complete responses, 19 partial responses, seven stable disease and two progressive disease, for an overall response rate of 72% (95% confidence interval 53% to 86%). Eight (25%) patients with residual liver or lung metastases were radically resected after chemotherapy. After a median follow-up of 18.1 months, the median progression-free survival is 10.8 months and median survival is 28.4 months. CONCLUSIONS: This simplified FOLFOXIRI combination can be delivered easily in outpatient settings, with manageable toxic effects, and has very promising antitumor activity. While the safety profile seems to be improved in comparison with our previous FOLFOXIRI regimen, antitumor activity and efficacy appear to be maintained. PMID- 15550582 TI - Irinotecan in combination with 5-fluorouracil and folinic acid or with cisplatin in patients with advanced gastric or esophageal-gastric junction adenocarcinoma: results of a randomized phase II study. AB - BACKGROUND: To identify the most effective of two combinations, irinotecan/5 fluorouracil (5-FU)/folinic acid (FA) and irinotecan/cisplatin, in the treatment of advanced gastric cancer, for investigation in a phase III trial. PATIENTS AND METHODS: Patients were randomized to receive irinotecan [80 mg/m2 intravenously (i.v.)], FA (500 mg/m2 i.v.) and a 22-h infusion of 5-FU (2000 mg/m2 i.v.), weekly for 6 weeks with a 1-week rest, or irinotecan (200 mg/m2 i.v.) and cisplatin (60 mg/m2 i.v.), on day 1 for 3 weeks. RESULTS: A total of 115 patients were eligible for analysis in the per-protocol population. The overall response rate in the irinotecan/5-FU/FA arm (n=59) was 42.4%, with a complete response rate of 5.1%. Corresponding figures for the irinotecan/cisplatin arm (n=56) were 32.1% and 1.8%, respectively. The median time to progression was 6.5 months (irinotecan/5-FU/FA) and 4.2 months (irinotecan/cisplatin) (P < 0.0001), with median survival times of 10.7 and 6.9 months, respectively (P=0.0018). The major toxicity was grade 3/4 neutropenia, which was more pronounced with irinotecan/cisplatin than with irinotecan/5-FU/FA (65.7% versus 27%). Diarrhea was the main grade 3/4 non-hematological toxicity with both irinotecan/5-FU/FA (27.0%) and irinotecan/cisplatin (18.1%). CONCLUSIONS: Both combinations were active, with acceptable safety profiles. Irinotecan/5-FU/FA was selected as the most effective combination for investigation in a phase III trial in advanced gastric cancer. PMID- 15550583 TI - Addition of platinum compounds to a new agent in patients with advanced non-small cell lung cancer: a literature based meta-analysis of randomised trials. AB - BACKGROUND: Single new agents reportedly produce promising response and survival effects, but platinum-based doublets remain the standard chemotherapy for advanced non-small-cell lung cancer (NSCLC). The aim of this study was to evaluate the effectiveness of platinum for advanced NSCLC by carrying out a meta analysis of trials that compared platinum-based doublets with single new agent therapy alone. METHODS: We carried out a literature search to identify trials, conducted between 1994 and 2003, comparing a doublet of platinum plus a new agent with a new agent alone in previously untreated patients with advanced NSCLC. Outcomes analysed were response, survival and toxicity. RESULTS: Eight trials encompassing 2374 patients were identified. Platinum-based doublets produced an approximately two-fold higher overall (complete and partial) response rate than the new agent alone [odds ratio = 2.32; 95% confidence interval (CI)=1.68-3.20]. Platinum-based doublet therapy was also associated with a 13% prolongation of survival (hazard ratio = 0.87; 95% CI = 0.80-0.94, P <0.001). Despite significant increases in the frequencies of various toxic effects in patients receiving platinum-based doublets, no significant difference in treatment-related mortality was observed. CONCLUSION: This is the first published meta-analysis demonstrating the importance of combining platinum with single new agents in the treatment of advanced NSCLC. PMID- 15550584 TI - All aggressive lymphoma subtypes do not share similar outcome after front-line autotransplantation: a matched-control analysis by the Groupe d'Etude des Lymphomes de l'Adulte (GELA). AB - BACKGROUND: Data are still conflicting on the indication of front-line autologous stem-cell transplantation (ASCT) as consolidation for aggressive lymphoma. To assess the therapeutic effect of ASCT among different aggressive lymphoma subtypes, we conducted a matched-control analysis by pooling the data from two Groupe d'Etude des Lymphomes de l'Adulte (GELA) trials. PATIENTS AND METHODS: Between October 1987 and September 1998, 330 patients received ASCT after achieving complete remission with the ACBVP induction regimen. The histological slides showed: B aggressive non-Hodgkin's lymphoma (B-NHL) in 249 patients (75%), T-NHL in 52 patients (15%) (including 23 T anaplastic) and non-classified NHL in 29 patients. The age-adjusted International Prognostic Index (aaIPI) was 2 or 3 in 66%. Patients were matched with controls from the same GELA database but treated with chemotherapy only. RESULTS: ASCT did not benefit non-anaplastic T NHL patients [5-year overall survival (OS) 44% (chemotherapy) versus 49% (ASCT), P=0.87; disease-free survival (DFS) 38% versus 45%, P=0.89] in comparison with B NHL [5-year OS 77% (chemotherapy) versus 79% (ASCT), P=0.64; DFS 67% versus 72%, P=0.13]. However, for B-NHL patients with aaIPI score 2 or 3, the benefit of ASCT was significant. CONCLUSIONS: This cohort study confirms the high efficacy of front-line ASCT in responding aggressive B-NHL patients with adverse prognostic factors. PMID- 15550585 TI - Treatment of stage I and II Hodgkin's lymphoma with ABVD chemotherapy: results after 7 years of a prospective study. AB - BACKGROUND: Chemotherapy with ABVD (doxorubicin, bleomycin, vinblastine and darcarbacine) schedule is the standard treatment for advanced Hodgkin's lymphoma. Certain facts, including a low toxicity compared with MOPP/ABV (mechlorethamine, vincristine, procarbazine, prednisone, doxorubicin, bleomycin and vinblastine) and minimal potential for inducing second neoplasias or patient sterility, support the use of ABVD to treat early disease stages. In the present study, we prospectively evaluated the long-term efficacy and toxicity of six cycles of ABVD as treatment for early-stage Hodgkin's lymphoma. PATIENTS AND METHODS: From January 1990 to June 2002, 95 patients with stage I and II Hodgkin's lymphoma were treated with six ABVD cycles. Fifteen patients who met the criteria for mediastinal bulky disease also received further radiotherapy on the mediastinum. RESULTS: After six cycles, 89 patients (94%) showed a complete response (CR) and six patients (6%) showed a partial response (PR). These PRs became CRs after radiotherapy. After a median follow-up of 78 months, 14 patients had relapsed and three had died. Overall survival and progression-free survival rates at 7 years were 96% and 84%, respectively. For patients with stage IA and IIA without mediastinal bulky disease, the survival rates were 97% and 88%, respectively. CONCLUSIONS: The administration of six ABVD cycles is an effective and safe treatment in patients with stage I and II Hodgkin's lymphoma. PMID- 15550586 TI - Effect of highly active antiretroviral therapy (HAART) on pharmacokinetics and pharmacodynamics of doxorubicin in patients with HIV-associated non-Hodgkin's lymphoma. AB - BACKGROUND: We demonstrated that highly active antiretroviral therapy (HAART) increases the toxic effect of cyclophosphamide, vincristine, doxorubicin (DOX) and prednisone (CHOP) in HIV-patients with non-Hodgkin's lymphoma (NHL). To ascertain the cause of increased toxicity, we investigated the pharmacokinetics of DOX in HIV-patients with NHL treated with CHOP with and without HAART. METHODS: Complete pharmacokinetics and pharmacodynamic analysis was determined in 19 patients during 38 cycles of chemotherapy: 19 cycles with CHOP and 19 CHOP + HAART in a crossover-designed study. HAART included protease inhibitors indinavir (IDV) in nine patients, saquinavir (SQV) hard gel in six patients and nelfinavir (NFV) in four patients. RESULTS: No significant effects of HAART on pharmacokinetics parameters of DOX were observed. Similarly, no differential effect on DOX pharmacokinetics among IDV, SQV, and NFV was evidenced. Significant associations (P=0.012) were observed between DOX AUC0-infinity (area under the concentration curve) and G3-G4 WHO haematologic toxicity, in patients treated with CHOP alone, but not in those treated with CHOP + HAART (P = not significant). CONCLUSION: We demonstrated that HAART therapy has no significant effect on DOX pharmacokinetics. DOX AUC appears to be a predictor of toxicity only in patients treated with CHOP alone. Other factors beside DOX plasma levels are detrimental for toxicity after CHOP + HAART. Therefore, pharmacodynamic interactions between HAART and DOX should be considered. PMID- 15550587 TI - Interferon-alpha-2b and oral cytarabine ocfosfate for newly diagnosed chronic myeloid leukaemia. AB - BACKGROUND: Treatment with interferon and subcutaneous cytarabine produces superior cytogenetic responses in chronic myeloid leukaemia (CML) than treatment with interferon alone, but at the expense of greater toxicity. Cytarabine ocfosfate (YNK01) is an oral precursor of cytarabine that may overcome some of the inconvenience and toxicities associated with subcutaneous cytarabine administration. PATIENTS AND METHODS: We studied the efficacy and tolerability of combination therapy with interferon-alpha-2b and YNK01 in patients with newly diagnosed, untreated CML. Forty patients were treated with interferon-alpha-2b (5 MU/m2/day) plus monthly courses of YNK01 (600 mg/day for 10 days) for 1 year. RESULTS: The 6-month complete haematological response rate was 63% and the 1-year major cytogenetic response rate was 30%, with 10% of cytogenetic responses being complete. With a median follow-up of 57 months, the estimated 5-year overall survival was 86% (95% confidence interval 70% to 94%). Treatment tolerability was poor, with toxicity leading to discontinuation of one or both drugs in 60% of cases. The median daily dose of interferon alpha-2b was 7.75 MU and the median dose of YNK01 was 600 mg/day for each 10-day treatment cycle. CONCLUSIONS: Interferon-alpha-2b and YNK01 produce cytogenetic responses comparable to those achieved with interferon-alpha-2b and parenteral cytarabine, although toxicity was excessive. Alternate dosing strategies may enhance the tolerability of YNK01. PMID- 15550589 TI - Phase I trial of a murine antibody to MUC1 in patients with metastatic cancer: evidence for the activation of humoral and cellular antitumor immunity. AB - BACKGROUND: BrevaRex mAb-AR20.5 is a murine anti-MUC1 monoclonal antibody generated to induce MUC1 antigen-specific immune responses through the formation of immune complexes with circulating MUC1 and/or MUC1-expressing tumor cells that may target these immune complexes (IC) to receptors on dendritic cells (DCs). PATIENTS AND METHODS: A phase I study focusing on safety and immunology evaluated 1, 2 and 4-mg doses. Seventeen patients with MUC1-positive cancers received intravenous infusions of the antibody over 30 min on weeks 1, 3, 5, 9, 13 and 17 of treatment. RESULTS: mAb-AR20.5 was well-tolerated, not associated with dose limiting toxicity, and did not induce hypersensitivity reactions. Overall, five of 15 evaluable patients developed human anti-mouse antibodies (HAMA), five developed anti-idiotypic antibodies (Ab2) and seven developed anti-MUC1 antibodies. Immune responses were most prominent in the 2-mg dose cohort for all parameters tested, and treatment-emergent MUC1-specific T-cell responses were detected in five of 10 evaluable patients treated with mAb-AR20.5. CONCLUSIONS: The injection of a murine antibody to MUC1 induces MUC1-specific immune responses in advanced cancer patients. Anti-MUC1 antibody increases correlated with decrease or stabilization of CA15.3 levels (P=0.03). The 2-mg dose of mAb-AR20.5 showed strongest biological activity, and will be evaluated in future efficacy trials. PMID- 15550588 TI - Phase I trial of intravenous aviscumine (rViscumin) in patients with solid tumors: a study of the European Organization for Research and Treatment of Cancer New Drug Development Group. AB - BACKGROUND: Aviscumine is an Escherichia coli-derived recombinant type II ribosome-inactivating protein with potent antitumor activity in vitro and in vivo. It is the recombinant counterpart of natural mistletoe lectin-I. The current study was performed to determine the safety profile, dose-limiting toxicity (DLT) and maximum tolerated dose (MTD) of the intravenous (i.v.) administration of aviscumine in cancer patients. Translational research included the evaluation of pharmacokinetics and monitoring of plasma cytokine and anti aviscumine antibody induction after administration of the drug. PATIENTS AND METHODS: Aviscumine was given twice weekly as a 1 h central i.v. infusion in patients with advanced, refractory progressive, solid malignant tumors who had not been previously exposed to natural mistletoe preparations. They had histologically or cytologically verified disease, were > or =18 years old, had an Eastern Cooperative Oncology Group performance status < or =2 and adequate bone marrow, liver and renal function. DLT was defined as any non-hematological grade 3-4 toxicity (National Cancer Institute Common Toxicity Criteria version 2.0), neutrophil count <500/microl for > or =7 days, febrile neutropenia or thrombocytopenia grade 4. The MTD was defined as the dose at which >20% of patients experienced DLT during the first treatment cycle. The Continual Reassessment Method was used to determine the number of patients required per dose level. RESULTS: Forty-one fully eligible patients (19 male, 22 female) with a median age of 56 years (range 37-74) were enrolled. Colorectal, ovarian, renal cell and breast cancer were the most common tumor types. Dose levels of aviscumine ranged from 10 to 6400 ng/kg. The median number of cycles was two (range one to eight). Common clinical toxicities in cycle 1 were fatigue, fever, nausea, vomiting and allergic reactions. Fatigue grade 3 was dose limiting in one of six patients at 4000 ng/kg and reversible grade 3 liver toxicity (elevation in alkaline phosphatase, transaminases and/or gamma-glutamyltransferase) occurred in one of 10 patients at 4800 ng/kg and in two of five patients at 6400 ng/kg. The best response (RECIST criteria) was stable disease in 11 patients, lasting for two to eight cycles. The pharmacokinetic evaluation revealed a short alpha half life of 13 min and linear kinetics on dose levels > or =1600 ng/kg. Aviscumine stimulated the immune system with a release of cytokines such as interleukin (IL) 1beta, IL-6 and interferon-gamma, and induced immunoglobulin (Ig) G- and/or IgM anti-aviscumine antibodies of uncertain clinical relevance. CONCLUSIONS: The recommended dose for further clinical trials is 5600 ng/kg twice weekly. Based on the short half-life of the recombinant protein observed in this trial, the exploration of prolonged infusion schedules of aviscumine is warranted. PMID- 15550590 TI - Risk of cancer at sites other than the breast in Swedish families eligible for BRCA1 or BRCA2 mutation testing. AB - BACKGROUND: Population-based data on the risk of cancer in families eligible for BRCA1/2 mutation testing may help to reach a consensus about the association of BRCA1/2 mutations with cancer at sites other than the breast and may reveal new, non-BRCA1/2 related components of the familial clustering of cancer in those families. PATIENTS AND METHODS: The families of the Swedish Family-Cancer Database with at least three generations (n = 944,723) were classified according to the criteria proposed by the German Consortium for Hereditary Breast and Ovarian Cancer. The cancer incidences in the classified families were compared with the incidences in the general population. The percentages of individuals with cancer in families eligible for BRCA1/2 mutation testing were compared with data in the literature to estimate the proportion of malignancies related to BRCA1/2 mutations. RESULTS: Families with two breast cancers before the age of 50 years showed increased risk of early onset pancreatic, prostate and ovarian cancers; families with ovarian and breast cancers presented increased incidences for ovarian and ocular cancers; families with two breast cancers, at least one of them under the age of 50 years, showed increased risks of prostate and primary liver cancers. Stomach cancer before age 70 years was twice as frequent in families with breast and ovarian cancers as in the general population. BRCA1/2 mutations probably explain most of the aggregation of ovarian cancer in families with male breast cancer, and in families with at least two breast cancers diagnosed before age 50 years. CONCLUSIONS: The association of BRCA1/2 mutations with ovarian, pancreatic, prostate and stomach cancers was confirmed at a population level. However, the clustering of early pancreatic cancer in families with two breast cancers under age 50 years, the aggregation of ovarian cancer in families with breast and ovarian cancers, and the increased incidence of early onset prostate cancer in families with male breast cancer seem to be due to other effects unrelated to BRCA1/2 mutations. PMID- 15550591 TI - Thyroid disease in northern Italian children born around the time of the Chernobyl nuclear accident. AB - BACKGROUND: The Chernobyl nuclear accident of 1986 caused a dramatic increase in the incidence of thyroid cancers in exposed children in Belarus. Airborne radioactivity from the reactor spread over northern Italy, where rainout gave rise to low levels of radioactivity at ground level. PATIENTS AND METHODS: As the latency between exposure to ionising radiation and development of thyroid cancer is thought to be about 10 years, in 1996/1997 all children born in 1985 and 1986 and attending school in an area of Milan, Italy were examined for thyroid nodules. A total of 3949 children were examined by two physicians blinded to the examination and diagnosis of the other. The children were to be reassessed in 2001/2002. RESULTS: In total, 1% had palpable nodules. The nodule diagnoses were: Hurtle cell adenoma (one), thyroglossal duct cyst (one), thyroid cyst (four) and thyroiditis (four). The prevalence of thyroid disease in the cohort was indistinguishable from that of populations not exposed to radioactive pollution. Only 10 children re-presented for examination 5 years later; all were negative. The direct costs of the study were estimated at 21,200 Euros. CONCLUSION: The high cost of the study in relation to reassuring lack of increase in thyroid nodule prevalence suggests that further studies are not justified. PMID- 15550592 TI - Zoledronic acid-associated thrombotic thrombocytopenic purpura. PMID- 15550593 TI - Solid tumors in patients treated for Hodgkin's disease. PMID- 15550594 TI - Symptomatic hypocalcaemia after treatment with zoledronic acid in a patient with multiple myeloma. PMID- 15550595 TI - Analysis of Bcl-2 and p53 protein expression in non-Hodgkin's lymphoma. PMID- 15550596 TI - CDX-2 should be included in the work-up of patients with lung metastases from unknown primary. PMID- 15550597 TI - An incidence density sampling program for nested case-control analyses. AB - BACKGROUND: The nested case-control design can be a very efficient approach to an epidemiological investigation. In order to obtain unbiased estimates of relative risk, controls should be selected by incidence density sampling, which involves matching each case to a sample of those who are at risk at the time of case occurrence. METHODS: This paper presents a simple computer program for incidence density sampling. This program was evaluated using data derived from a cohort study of mortality among workers employed in the nuclear weapons industry. Controls were selected for cases via incidence density sampling; an estimate of the exposure-mortality association was obtained via conditional logistic regression. After 100 iterations of this procedure, the average effect estimate was compared to the risk estimate obtained via proportional hazards regression. The same methods were used to evaluate a program for incidence density sampling that was proposed previously by Pearce in 1989. RESULTS: Relative risk estimates obtained from nested case-control analyses conducted using the incidence density sampling program reported in this paper are unbiased. In contrast, the program for incidence density sampling proposed by Pearce tended to produce biased relative risk estimates; the magnitude of bias increased with increasing numbers of controls selected per case. CONCLUSIONS: The computer program described in this paper offers a simple approach to incidence density sampling for nested case control analyses with exact matching on attained age and appropriate enumeration of the pool of eligible controls for each case. This method overcomes problems of bias inherent in a previously proposed program for incidence density sampling. PMID- 15550598 TI - Case-crossover designs in occupational health. PMID- 15550599 TI - Work related neck pain: how important is it, and how should we understand its causes? PMID- 15550600 TI - The effects of particulate air pollution on daily deaths: a multi-city case crossover analysis. AB - BACKGROUND: Numerous studies have reported that day-to-day changes in particulate air pollution are associated with day-to-day changes in deaths. Recently, several reports have indicated that the software used to control for season and weather in some of these studies had deficiencies. AIMS: To investigate the use of the case-crossover design as an alternative. METHODS: This approach compares the exposure of each case to their exposure on a nearby day, when they did not die. Hence it controls for seasonal patterns and for all slowly varying covariates (age, smoking, etc) by matching rather than complex modelling. A key feature is that temperature can also be controlled by matching. This approach was applied to a study of 14 US cities. Weather and day of the week were controlled for in the regression. RESULTS: A 10 microg/m3 increase in PM10 was associated with a 0.36% increase in daily deaths from internal causes (95% CI 0.22% to 0.50%). Results were little changed if, instead of symmetrical sampling of control days the time stratified method was applied, when control days were matched on temperature, or when more lags of winter time temperatures were used. Similar results were found using a Poisson regression, but the case-crossover method has the advantage of simplicity in modelling, and of combining matched strata across multiple locations in a single stage analysis. CONCLUSIONS: Despite the considerable differences in analytical design, the previously reported associations of particles with mortality persisted in this study. The association appeared quite linear. Case-crossover designs represent an attractive method to control for season and weather by matching. PMID- 15550601 TI - Psychosocial work factors and dementia. AB - AIMS: To evaluate the association between psychosocial workplace factors and dementia in a case-control study. METHODS: Patients with dementia (aged 55-99 years) were recruited from 23 general practices in the city of Frankfurt-on-Main and surrounding area in 1998-2000. Of these, 108 were suffering from possible Alzheimer's disease, 59 from possible vascular dementia, and 28 from secondary or unclassified dementia. A total of 229 control subjects (aged 60-94 years) was recruited from the same study region: 122 population controls and 107 dementia free ambulatory patients. A detailed job history was elicited in a structured personal interview (next-of-kin interviews of cases). Psychosocial work exposure was assigned to cases and control subjects by linking lifetime job histories with a Finnish job-exposure matrix. Data were analysed using logistic regression, to control for age, region, sex, dementia in parents, education, smoking, and the psychosocial network at age 30. RESULTS: There were decreased odds ratios for high challenge at work, high control possibilities at work, and high social demands at work. High risks for error at work revealed a significant positive association with the diagnosis of dementia. Restriction of the analysis to cases with possible Alzheimer's disease or to cases with possible vascular dementia led to similar results. CONCLUSIONS: These results support a role for psychosocial work factors in the aetiology of dementia. As an alternative explanation, people might have chosen jobs with poor work factors due to preclinical dementia, which becomes clinically manifest decades later. PMID- 15550602 TI - Dose-response relations between occupational exposures to physical and psychosocial factors and the risk of low back pain. AB - AIMS: To assess dose-response relations between occupational exposures to physical and psychosocial factors and the risk of low back pain. METHODS: A cohort of 523 subjects, working in nursing homes and homes for the elderly, was followed prospectively for one year. Physical load for different occupations was assessed by quantitative observations at the workplace. Information on low back pain and other factors was gathered with questionnaires administered at baseline and at one year. Two outcome measures of low back pain incidence were used: any new episode of pain lasting for at least a few hours during follow up (LBP); and any new episode of disabling pain that interfered with daily activities during follow up (LBP/D). Hierarchical regression analysis with a spline function was used to estimate dose-response relations. RESULTS: The risk of LBP was not associated with physical factors, controlling for confounders; but this outcome was inversely associated with age and weakly, though imprecisely, associated with two psychosocial factors--low decision authority and high work demands. In contrast, the risk of LBP/D was positively associated with age and not associated with the psychosocial factors. Trunk flexion over 45 degrees was monotonically associated with the risk of LBP/D; the estimated relative risk was 3.18 (95% CI 1.13 to 9.00) for 1 hour and 45 minutes of bending per week (90th centile), relative to 30 minutes per week. The hierarchical estimates of effect were more stable than were the maximum likelihood estimates. CONCLUSION: Occupational exposure to trunk flexion over 45 degrees appears to be a risk factor for low back pain with disability among persons employed in nursing homes and homes for the elderly in the Netherlands. PMID- 15550603 TI - Internal consistency and validity of a new physical workload questionnaire. AB - AIMS: To examine the dimensionality, internal consistency, and construct validity of a new physical workload questionnaire in employees with musculoskeletal complaints. METHODS: Factor analysis was applied to the responses in three study populations with musculoskeletal disorders (n = 406, 300, and 557) on 26 items related to physical workload. The internal consistency of the resulting subscales was examined. It was hypothesised that physical workload would vary among different occupational groups. The occupations of all subjects were classified into four groups on the basis of expected workload (heavy physical load; long lasting postures and repetitive movements; both; no physical load). Construct validity of the subscales created was tested by comparing the subscale scores among these occupational groups. RESULTS: The pattern of the factor loadings of items was almost identical for the three study populations. Two interpretable factors were found: items related to heavy physical workload loaded highly on the first factor, and items related to static postures or repetitive work loaded highly on the second factor. The first constructed subscale "heavy physical work" had a Cronbach's alpha of 0.92 to 0.93 and the second subscale "long lasting postures and repetitive movements", of 0.86 to 0.87. Six of eight hypotheses regarding the construct validity of the subscales were confirmed. CONCLUSIONS: The results support the internal structure, internal consistency, and validity of the new physical workload questionnaire. Testing this questionnaire in non symptomatic employees and comparing its performance with objective assessments of physical workload are important next steps in the validation process. PMID- 15550604 TI - Time trends in the incidence of myocardial infarction among professional drivers in Stockholm 1977-96. AB - AIMS: To investigate time trends in the incidence of first myocardial infarction (MI) among bus, taxi, and lorry drivers in Stockholm. METHODS: In this population based case-control study, all first events of acute MI among men aged 40-69 in Stockholm County 1977-96 were identified using registers of hospital discharges and deaths. Controls were selected randomly from the general population. National censuses were used for information on occupational titles. A total of 20,364 cases and 136,342 controls were included in the study. Among these, 1183 cases and 6072 controls had worked as drivers. RESULTS: During 1977-84 the MI incidence was higher in all three driver groups than among other manual workers. There was a decline in MI incidence among drivers as well as in the general population during the study period. The decline was greater among drivers than among other manual workers. During 1985-96 the relative risk remained increased compared to other manual workers among taxi and lorry drivers, but not among bus drivers. CONCLUSIONS: Despite a reduced MI incidence for professional drivers in recent years, preventive measures to reduce the risk of MI in these occupational groups, and in particular among taxi and lorry drivers, continue to be of importance. PMID- 15550605 TI - Occupational and personal factors associated with acquired lumbar spondylolisthesis of urban taxi drivers. AB - AIMS: To investigate the occupational and personal factors associated with lumbar spondylolisthesis in taxi drivers. METHODS: Cross-sectional analysis of the baseline data from the Taxi Drivers' Health Study cohort. Information was retrieved from the medical records of standardised lumbosacral spine plain films, age, and anthropometric measures of 1242 subjects. Acquired spondylolisthesis (ASL) was defined as non-lytic spondylolisthesis involving lumbar spines above L5. Questionnaires were used to gather information on demographic features, health behaviours, exercise, work related physical and psychosocial factors, and driving time profiles. Multiple logistic regression was used to model the odds ratio (OR) for prevalent ASL cases associated with personal and occupational factors. RESULTS: A total of 40 cases (3.2%) of ASL were diagnosed. Among those driving < or =5 years, 6-15 years, and >15 years, the estimated prevalence of lumbar spondylolisthesis was 1.1%, 2.4%, and 7.1% respectively. Results of multiple logistic regression suggested that taxicab driving >15 years (OR = 3.4, 95% CI 1.1 to 10.7, compared to driving < or =5 years), age (OR = 2.6, 95% CI 1.1 to 6.6 for age 46-55; and OR = 4.8, 95% CI 1.8 to 12.9 for age >55), body mass index > or =25 kg/m2 (OR = 2.2, 95% CI 1.1 to 4.6), and frequent strenuous exercise (OR = 2.2, 95% CI 1.1 to 4.5) were significantly associated with higher prevalence of spondylolisthesis. There was a consistent likely exposure-response relation between professional seniority and ASL prevalence. CONCLUSIONS: Longitudinal studies are needed to confirm the observed association between professional driving and spondylolisthesis, and to examine further the specific occupational exposures accountable for this association. PMID- 15550606 TI - Study on the relation between occupational fenvalerate exposure and spermatozoa DNA damage of pesticide factory workers. AB - AIMS: To determine sperm nuclear DNA integrity and to investigate the relation between fenvalerate (FE) exposure and spermatozoa DNA damage. METHODS: Sperm DNA fragmentation was detected by a modified alkaline single cell gel electrophoresis (Comet) assay and a terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) assay. The olive tail moment (OTM) and percentage tail DNA were measured by the Comet assay, and cell positive percentage was measured by the TUNEL assay for DNA damage evaluation. RESULTS: The DNA integrity of spermatozoa of external and internal control groups were both significantly greater than that of the FE exposed group. The median value of tail DNA percentage in the exposure group was 11.30, which was significantly higher than 5.60 in the internal control group and 5.10 in the external control group. The median value of OTM was 3.80 in the exposure group, significantly higher than 1.50 in the internal control group and 2.00 in the external control group. Mean cell positive was 31.2% in the exposure group, significantly higher than 17.4% in the internal control and 19.6% in the external control groups. Cell positive (%) was significantly correlated with tail DNA percentage and with OTM of whole subjects (n = 63). CONCLUSIONS: Results showed that occupational FE exposure is associated with an increase in sperm DNA damage. A combination of the Comet and TUNEL assays would offer more comprehensive information for a better understanding of sperm DNA damage, and the biological significance of sperm DNA damage in sperm function and male infertility. PMID- 15550607 TI - Symptoms and medical conditions in Australian veterans of the 1991 Gulf War: relation to immunisations and other Gulf War exposures. AB - AIMS: To investigate whether Australian Gulf War veterans have a higher than expected prevalence of recent symptoms and medical conditions that were first diagnosed in the period following the 1991 Gulf War; and if so, whether these effects were associated with exposures and experiences that occurred in the Gulf War. METHODS: Cross-sectional study of 1456 Australian Gulf War veterans and a comparison group who were in operational units at the time of the Gulf War, but were not deployed to that conflict (n = 1588). A postal questionnaire was administered and the likelihood of the diagnosis of self-reported medical conditions was assessed and rated by a medical practitioner. RESULTS: Gulf War veterans had a higher prevalence of all self-reported health symptoms than the comparison group, and more of the Gulf War veterans had severe symptoms. Increased symptom reporting was associated with several exposures, including having more than 10 immunisations, pyridostigmine bromide tablets, anti biological warfare tablets, pesticides, insect repellents, reportedly being in a chemical weapons area, and stressful military service experiences in a strong dose-response relation. Gulf War veterans reported psychological (particularly post-traumatic stress disorder), skin, eye, and sinus conditions first diagnosed in 1991 or later more commonly than the comparison group. Over 90% of medical conditions reported by both study groups were rated by a medical practitioner as having a high likelihood of diagnosis. CONCLUSION: More than 10 years after the 1991 Gulf War, Australian veterans self-report all symptoms and some medical conditions more commonly than the comparison group. Further analysis of the severity of symptoms and likelihood of the diagnosis of medical conditions suggested that these findings are not due to over-reporting or to participation bias. PMID- 15550608 TI - The health of Australian veterans of the 1991 Gulf War: factor analysis of self reported symptoms. AB - BACKGROUND: A recent report showed that Australian veterans of the 1991 Gulf War displayed a greater prevalence of a multitude of self-reported symptoms than a randomly sampled comparison group of military personnel who were eligible for deployment but were not deployed to the Gulf. AIMS: To investigate whether the pattern, rather than frequency, of symptom reporting in these Australian Gulf War veterans differed from that of the comparison group personnel. METHODS: Factor analysis was used to determine whether the co-occurrence of 62 symptoms in 1322 male Gulf War veterans can be explained by a number of underlying dimensions, called factors. The methodology was also applied to 1459 male comparison group subjects and the factor solutions of the two groups were compared. RESULTS: For the Gulf War veterans, a three factor solution displayed replicability and construct validity. The three factors were labelled as psycho-physiological distress, somatic distress, and arthro-neuromuscular distress, and were broadly similar to those described in previous studies of Gulf War veterans. A concordant three factor solution was also found for the comparison group subjects, with strong convergence of the factor loadings and factor scores across the two groups being displayed. CONCLUSION: Results did not display evidence of a unique pattern of self-reported symptoms among Gulf War veterans. Results also indicated that the differences between the groups lie in the degrees of expression of the three underlying factors, consistent with the well documented evidence of increased self-reported symptom prevalence in Gulf War veterans. PMID- 15550609 TI - Stress and dysmenorrhoea: a population based prospective study. AB - BACKGROUND: Dysmenorrhoea is the most common gynaecological disorder in women of reproductive age. Despite the association between stress and pregnancy outcomes, few studies have examined the possible link between stress and dysmenorrhoea. AIMS AND METHODS: Using a population based cohort of Chinese women, the independent effect of women's perceived stress in the preceding menstrual cycle on the incidence of dysmenorrhoea in the subsequent cycle was investigated prospectively. The analysis included 1160 prospectively observed menstrual cycles from 388 healthy, nulliparous, newly married women who intended to conceive. The perception of stress and the occurrence of dysmenorrhoea in each menstrual cycle were determined from daily diaries recorded by the women. RESULTS: After adjustment for important covariates, the risk of dysmenorrhoea was more than twice as great among women with high stress compared to those with low stress in the preceding cycle (OR = 2.4; 95% CI 1.4 to 4.3). The risk of dysmenorrhoea was greatest among women with both high stress and a history of dysmenorrhoea compared to women with low stress and no history of dysmenorrhoea (OR = 10.4, 95% CI 4.9 to 22.3). Stress in the follicular phase of the preceding cycles had a stronger association with dysmenorrhoea than stress in the luteal phase of the preceding cycles. CONCLUSION: This study shows a significant association between stress and the incidence of dysmenorrhoea, which is even stronger among women with a history of dysmenorrhoea. PMID- 15550610 TI - A comparison of methods for the assessment of postural load and duration of computer use. AB - AIM: To compare two different methods for assessment of postural load and duration of computer use in office workers. METHODS: The study population existed of 87 computer workers. Questionnaire data about exposure were compared with exposures measured by a standardised or objective method. Measuring true exposure to postural load consisted of an observation of the workstation design and posture by a trained observer. A software program was used to record individual computer use. RESULTS: Comparing the answers for each item of postural load, six of eleven items showed low agreement (kappa <0.20). For six items the sensitivity was below 50%, while for eight items the specificity was 80% or higher. Computer workers were unable to identify risk factors in their workplace and work posture. On average, computer workers overestimated their total computer use by 1.6 hours. The agreement among employees who reported a maximum of three hours of computer use per day was higher than the agreement among employees with a high duration of computer use. CONCLUSIONS: Self-report by means of this questionnaire is not a very reliable method to measure postural load and duration of computer use. This study emphasises that the challenge to develop quick and inexpensive techniques for assessing exposure to postural load and duration of computer use is still open. PMID- 15550611 TI - Leisure time physical activity and strenuousness of work as predictors of physical functioning: a 28 year follow up of a cohort of industrial employees. AB - AIMS: To examine associations of leisure time physical activity and physical strenuousness of work with physical functioning 28 years later. METHODS: A cohort (n = 902) of metal industry employees was studied for exercise and housework activity in 1973 and 1978, and for BMI, current smoking, strenuousness of work, grip strength, and chronic diseases in 1973. Of the 670 survivors in 2000, 529 (79%) responded to all studied items in a follow up questionnaire including the SF-36 Physical Functioning (PF) scale. Belonging to the lower quartile of the PF scale denoted poor functioning. RESULTS: Vigorous exercise and housework activity were inversely associated with poor PF 28 years later in both white-collar and blue-collar workers. Engaging in activities of any intensity was similarly associated among the blue-collar workers. In a multiple logistic regression model including as independent variables age, sex, occupational class, the number of chronic diseases, vigorous leisure time physical activity, BMI, physical work strenuousness, and smoking (all measured at baseline), the risk of poor PF at follow up was decreased by vigorous leisure time physical activity and increased by high physical strenuousness of work, high BMI, and smoking. The effect of work strenuousness was mainly due to that among the blue-collar group. Allowing for baseline grip strength did not materially alter the results. CONCLUSION: Vigorous leisure time physical activity decreased the risk of poor physical functioning as perceived considerably later in life, while high work strenuousness, smoking, and overweight increased it. Among blue-collar workers a beneficial association was observed with all leisure time activity, including that of lower intensity. PMID- 15550612 TI - Social environment modulates photoperiodic immune and reproductive responses in adult male white-footed mice (Peromyscus leucopus). AB - Social cues may interact with photoperiod to regulate seasonal adaptations in photoperiod-responsive rodents. Specifically, photoperiod-induced adjustments (e.g., reproduction and immune function) may differ among individuals in heterosexual pairs, same-sex pairs, or isolation. Heterosexual cues may be more influential, based on their potential fitness value, than same-sex cues or no social cues. The present study examined the effects of pair (with a male or female) or individual housing on reproductive and immune responses in male white footed mice (Peromyscus leucopus) maintained in long or short photoperiods. Female pairing did not affect reproductive responses in short-day males. In long days, however, the presence of a female increased both testosterone concentrations and testes mass compared with individually housed and male-paired mice, respectively. Short-day, individually housed males enhanced delayed-type hypersensitivity (DTH) responses compared with single-housed mice in long days, but all paired groups decreased DTH responses regardless of photoperiod. The lack of enhanced DTH response in male mice paired with females coincided with reduced circulating corticosterone concentrations in both photoperiod treatments. Together, these results suggest that social environment may have important modulatory effects on photoperiod-regulated immune responses in male white-footed mice. PMID- 15550613 TI - Sensory or sympathetic white adipose tissue denervation differentially affects depot growth and cellularity. AB - Functional and histological evidence for the sympathetic nervous system (SNS) innervation of white adipose tissue (WAT) exists for several species; however, its sensory innervation has only been shown in laboratory rats, and its function is unclear. We tested the effects of sensory and SNS innervation of Siberian hamster epididymal and inguinal WAT (EWAT and IWAT) by assessing calcitonin gene related peptide (CGRP)- and tyrosine hydroxylase-immunoreactivity (ir), respectively. Next, we tested the role of the sensory innervation of WAT on growth and cellularity because WAT surgical denervation increases pad mass via selective increases in fat cell number, an effect ascribed to SNS denervation but that could be due to the accompanying surgical disruption of WAT sensory innervation. Sensory denervation was accomplished via multiple local microinjections of capsaicin into WAT, and its effects were compared with those of surgical denervation. Surgically denervated IWAT and EWAT showed significantly decreased tyrosine hydroxylase-ir and CGRP-ir, whereas capsaicin-treated WAT had only significantly decreased CGRP-ir. Surgically denervated pad masses were significantly increased; this was accompanied by increased total fat cell number in IWAT, with no change in fat cell size. EWAT only showed a significant increase in the number of small- to medium-sized adipocytes (75-125 mum diameter). By contrast, sensory-denervated pad masses were unchanged, but IWAT showed significantly increased average fat cell size. Collectively, these data provide immunohistochemical evidence for sensory and SNS innervation of WAT in Siberian hamsters and differential control of WAT cellularity by these innervations, as well as the ability of locally applied capsaicin to selectively reduce WAT sensory innervation. PMID- 15550614 TI - 17beta-estradiol downregulates tissue angiotensin-converting enzyme and ANG II type 1 receptor in female rats. AB - Estrogens have been implicated in both worsening and protecting from cardiovascular disease. The effects of 17beta-estradiol (E2) on the cardiovascular system may be mediated, at least in part, by its modulation of local tissue renin-angiotensin systems (RAS). We assessed two critical components, angiotensin-converting enzyme (ACE) and ANG II type 1 receptor (AT(1)R), in the heart, lung, abdominal aorta, adrenal, kidney, and brain in four groups of female Wistar rats (n = 5-6/group): 1) sham ovariectomized, 2) ovariectomized (OVX) treated with subcutaneous vehicle, 3) OVX treated with 25 mug/day (regular) E2 subcutaneously, and 4) OVX treated with 250 mug/day (high) subcutaneous E2 for 2 or 5 wk. After 2 wk, plasma ACE activity was not altered by OVX, but it was 34-38% lower in OVX + regular E2 and OVX + high E2 rats compared with sham OVX rats, and these decreases were no longer present after 5 wk. After 5 wk, OVX alone increased ACE activity and binding densities, and AT(1)R binding densities by 15-100% in right ventricle, left ventricle (LV), kidney, lung, abdominal aorta, adrenal and several cardiovascular regulatory nuclei in the brain. These effects were, for the most part, prevented by regular E2 replacement and were reversed to decreases by high E2 treatment. This regulation of tissue ACE and AT(1)R is significant as the activity of these tissue RAS contributes to the pathogenesis and/or progression of hypertension, atherosclerosis, and LV remodeling after myocardial infarction. PMID- 15550615 TI - Food entrainment modifies the c-Fos expression pattern in brain stem nuclei of rats. AB - When food is restricted to a few hours daily, animals increase their locomotor activity 2-3 h before food access, which has been termed food anticipatory activity. Food entrainment has been linked to the expression of a circadian food entrained oscillator (FEO) and the anatomic substrate of this oscillator seems to depend on diverse neural systems and peripheral organs. Previously, we have described a differential involvement of hypothalamic nuclei in the food-entrained process. For the food entrainment pathway, the communication between the gastrointestinal system and central nervous system is essential. The visceral synaptic input to the brain stem arrives at the dorsal vagal complex and is transmitted directly from the nucleus of the solitary tract (NST) or via the parabrachial nucleus (PBN) to hypothalamic nuclei and other areas of the forebrain. The present study aims to characterize the response of brain stem structures in food entrainment. The expression of c-Fos immunoreactivity (c-Fos IR) was used to identify neuronal activation. Present data show an increased c Fos-IR following meal time in all brain stem nuclei studied. Food-entrained temporal patterns did not persist under fasting conditions, indicating a direct dependence on feeding-elicited signals for this activation. Because NST and PBN exhibited a different and increased response from that expected after a regular meal, we suggest that food entrainment promotes ingestive adaptations that lead to a modified activation in these brain stem nuclei, e.g., stomach distension. Neural information provided by these nuclei to the brain may provide the essential entraining signal for FEO. PMID- 15550616 TI - Activation of slowly conducting medullary raphe-spinal neurons, including serotonergic neurons, increases cutaneous sympathetic vasomotor discharge in rabbit. AB - Neurons in the rostral medullary raphe/parapyramidal region regulate cutaneous sympathetic nerve discharge. Using focal electrical stimulation at different dorsoventral raphe/parapyramidal sites in anesthetized rabbits, we have now demonstrated that increases in ear pinna cutaneous sympathetic nerve discharge can be elicited only from sites within 1 mm of the ventral surface of the medulla. By comparing the latency to sympathetic discharge following stimulation at the ventral raphe site with the corresponding latency following stimulation of the spinal cord [third thoracic (T3) dorsolateral funiculus] we determined that the axonal conduction velocity of raphe-spinal neurons exciting ear pinna sympathetic vasomotor nerves is 0.8 +/- 0.1 m/s (n = 6, range 0.6-1.1 m/s). Applications of the 5-hydroxytryptamine (HT)(2A) antagonist trans-4-((3Z)3-[(2 dimethylaminoethyl)oxyimino]-3-(2-fluorophenyl)propen-1-yl)-phenol, hemifumarate (SR-46349B, 80 microg/kg in 0.8 ml) to the cerebrospinal fluid above thoracic spinal cord (T1-T7), but not the lumbar spinal cord (L2-L4), reduced raphe-evoked increases in ear pinna sympathetic vasomotor discharge from 43 +/- 9 to 16 +/- 6% (P < 0.01, n = 8). Subsequent application of the excitatory amino acid (EAA) antagonist kynurenic acid (25 micromol in 0.5 ml) substantially reduced the remaining evoked discharge (22 +/- 8 to 6 +/- 6%, P < 0.05, n = 5). Our conduction velocity data demonstrate that only slowly conducting raphe-spinal axons, in the unmyelinated range, contribute to sympathetic cutaneous vasomotor discharge evoked by electrical stimulation of the medullary raphe/parapyramidal region. Our pharmacological data provide evidence that raphe-spinal neurons using 5-HT as a neurotransmitter contribute to excitation of sympathetic preganglionic neurons regulating cutaneous vasomotor discharge. Raphe-spinal neurons using an EAA, perhaps glutamate, make a substantial contribution to the ear sympathetic nerve discharge evoked by raphe stimulation. PMID- 15550617 TI - Glucose does not activate nonadrenergic, noncholinergic inhibitory neurons in the rat stomach. AB - We reported previously that intravenously administered d-glucose acts in the central nervous system to inhibit gastric motility induced by hypoglycemia in anesthetized rats. The purpose of this study was to determine whether this effect is due to inhibition of dorsal motor nucleus of the vagus (DMV) cholinergic motoneurons, which synapse with postganglionic cholinergic neurons, or to excitation of DMV cholinergic neurons, which synapse with postganglionic nonadrenergic, noncholinergic (NANC) neurons, particularly nitrergic neurons. Three approaches were employed: 1) assessment of the efficacy of d-glucose induced inhibition of gastric motility in hypoglycemic rats with and without inhibition of nitric oxide synthase [10 mg/kg iv nitro-l-arginine methyl ester (l NAME)], 2) assessment of the efficacy of intravenous bethanechol (30 mug.kg( 1).min(-1)) to stimulate gastric motility in hypoglycemic rats during the time of d-glucose-induced inhibition of gastric motility, and 3) determination of c-Fos expression in DMV neurons after intravenous d-glucose was administered to normoglycemic rats. Results obtained demonstrated that l-NAME treatment had no effect on d-glucose-induced inhibition of gastric motility; there was no reduction in the efficacy of intravenous bethanechol to increase gastric motility, and c-Fos expression was not induced by d-glucose in DMV neurons that project to the stomach. These findings indicate that excitation of DMV cholinergic motoneurons that synapse with postganglionic NANC neurons is not a significant contributing component of d-glucose-induced inhibition of gastric motility. PMID- 15550618 TI - Central angiotensin II AT1 receptors mediate fetal swallowing and pressor responses in the near-term ovine fetus. AB - Swallowed volumes in the fetus are greater than adult values (per body weight) and serve to regulate amniotic fluid volume. Central ANG II stimulates swallowing, and nonspecific ANG II receptor antagonists inhibit both spontaneous and ANG II-stimulated swallowing. In the adult rat, AT1 receptors mediate both stimulated drinking and pressor activities, while the role of AT2 receptors is controversial. As fetal brain contains increased ANG II receptors compared with the adult brain, we sought to investigate the role of both AT1 and AT2 receptors in mediating fetal swallowing and pressor activities. Five pregnant ewes with singleton fetuses (130 +/- 1 days) were prepared with fetal vascular and lateral ventricle (LV) catheters and electrocorticogram and esophageal electromyogram electrodes and received three studies over 5 days. On day 1 (ANG II), following a 2-h basal period, 1 ml artificial cerebrospinal fluid (aCSF) was injected in the LV. At time 4 h, ANG II (6.4 microg) was injected in the LV, and the fetus was monitored for a final 2 h. On day 3, AT1 receptor blocker (losartan 0.5 mg) was administered at 2 h, and ANG II plus losartan was administered at 4 h. On day 5, AT2 receptor blocker (PD-123319; 0.8 mg was administered at 2 h and ANG II plus PD-123319 at 4 h. In the ANG II study, LV injection of ANG II significantly increased fetal swallowing (0.9 +/- 0.1 to 1.4 +/- 0.1 swallows/min; P < 0.05). In the losartan study, basal fetal swallowing significantly decreased in response to blockade of AT1 receptors (0.9 +/- 0.1 to 0.4 +/- 0.1 swallows/min; P < 0.05), while central injection of ANG II in the presence of AT1 receptor antagonism did not increase fetal swallowing (0.6 +/- 0.1 swallows/min). In the PD-123319 study, basal fetal swallowing did not change in response to blockade of AT2 receptor (0.9 +/- 0.1 swallows/min), while central injection of ANG II in the presence of AT2 blockade significantly increased fetal swallowing (1.5 +/- 0.1 swallows/min; P < 0.05). ANG II caused significant pressor responses in the control and PD 123319 studies but no pressor response in the presence of AT1 blockade. These data demonstrate that in the near-term ovine fetus, AT1 receptor but not AT2 receptors accessible via CSF contribute to dipsogenic and pressor responses. PMID- 15550619 TI - Adult sterol metabolism is not affected by a positive sterol balance in the neonatal Golden Syrian hamster. AB - Dietary components impact metabolism early in life. Some of the diet-induced effects are long lasting and can lead to various adult-based diseases. In the current studies, we examined the short-term effects of dietary cholesterol on neonatal hepatic sterol metabolism and the long-term effects that those early life diets had on sterol metabolism in adulthood. Neonatal hamsters began consuming solid food as a supplement to milk by 5 days of age; diets contained 0 or 2% added cholesterol (wt/wt). By 10 days of age, plasma and liver cholesterol concentrations were 3.2- and 2.5-fold greater, respectively, in the neonates fed cholesterol. Hepatic sterol synthesis rates were suppressed 65% in cholesterol fed neonates compared with control neonates. By 20 days of age, plasma and liver cholesterol concentrations were still greater and sterol synthesis rates were now suppressed maximally in neonates fed cholesterol compared with control neonates. The expression level of an apolipoprotein B-containing lipoprotein receptor (low density lipoprotein receptor-related protein) was greater and the mature form of the sterol regulatory element-binding protein-2 was similar in livers of 20-day old control neonates compared with control neonates at 10 days of age. To test whether the change in sterol balance in the neonatal period had a lasting effect on hepatic sterol metabolism, all animals were weaned on a low-cholesterol diet. At 70 days of age, hepatic sterol synthesis rates, plasma lipoprotein and liver cholesterol concentrations, and bile acid pool sizes and compositions were measured. Sterol balance in the adults was similar between animals fed either diet early in life, as demonstrated by a lack of difference in any parameter measured. Thus, even though dietary cholesterol suppressed hepatic sterol synthesis rates dramatically in the neonatal hamster, the change has little impact on sterol balance later in life. PMID- 15550620 TI - Blood pressure maintenance in NHE3-deficient mice with transgenic expression of NHE3 in small intestine. AB - NHE3 Na(+)/H(+) exchanger knockout (Nhe3(-/-)) mice have severe absorptive deficits in the kidney proximal tubule and intestinal tract. The resulting hypovolemia has confounded efforts to carefully evaluate the specific effects of NHE3 deficiency on kidney function. Development of mice with transgenic expression of NHE3 in the small intestine (tgNhe3(-/-)) has allowed us to analyze the role of renal NHE3 in overall maintenance of blood pressure, pressure natriuresis, and autoregulation of both glomerular filtration rate (GFR) and renal blood flow (RBF). Ambulatory blood pressure, measured by telemetry, was lower in tgNhe3(-/-) mice than in wild-type controls (tgNhe3(+/+)) when the mice were maintained on a normal NaCl diet but was normalized when they were provided with a high NaCl intake. Furthermore, administration of the AT1-receptor blocker losartan showed that circulating ANG II plays a major role in maintaining blood pressure in tgNhe3(-/-) mice fed normal NaCl but not in those receiving high NaCl. Clearance studies revealed a blunted pressure-natriuresis response in tgNhe3(-/-) mice at lower blood pressures but a robust response at higher blood pressures. Autoregulation of GFR and RBF was normal in tgNhe3(-/-) mice. These results show that dietary NaCl loading normalizes blood pressure in awake tgNhe3( /-) mice and that alterations in NHE3 activity are not essential for normal autoregulation of GFR and RBF. Furthermore, the data strongly support the hypothesis that NHE3 plays an important role in the diuretic and natriuretic responses to increases in blood pressure but also show that mechanisms not involving NHE3 mediate pressure natriuresis in the higher range of blood pressures studied. PMID- 15550621 TI - CCK inhibits the orexigenic effect of peripheral ghrelin. AB - CCK and ghrelin exert antagonistic effects on ingestive behavior. The aim of the present study was to investigate the interaction between ghrelin and CCK administered peripherally on food intake and neuronal activity in specific hypothalamic and brain stem nuclei, as assessed by c-Fos-like immunoreactivity (c FLI) in nonfasted rats. Ghrelin (13 microg/kg body wt) injected intraperitoneally significantly increased the cumulative food intake when measured at 30 min and 1 h after injection, compared with the vehicle group (2.9 +/- 1.0 g/kg body wt vs. 1.2 +/- 0.5 g/kg body wt, P < 0.028). Sulfated CCK octapeptide (CCK-8S) (2 or 25 microg/kg body wt) injected simultaneously blocked the orexigenic effect of ghrelin (0.22 +/- 0.13 g/kg body wt, P < 0.001 and 0.33 +/- 0.23 g/kg body wt, P < 0.0008), while injected alone, both doses of CCK-8S exerted a nonsignificant trend to reduce food intake. Ghrelin (13 microg/kg body wt ip) markedly increased the number of c-FLI-positive neurons per section in the arcuate nucleus (ARC) compared with vehicle (median: 31.35 vs. 9.86, P < 0.0001). CCK-8S (2 or 25 microg/kg body wt ip) had no effect on neuronal activity in the ARC, as assessed by c-FLI (median: 5.33 and 11.21 cells per section), but blocked the ghrelin induced increase of c-fos expression in this area when both peptides were administered simultaneously (median: 13.33 and 12.86 cells per section, respectively). Ghrelin at this dose had no effect on CCK-induced stimulation of c fos expression in the paraventricular nucleus of the hypothalamus and the nucleus of the solitary tract. These results suggest that CCK abolishes ghrelin-induced food intake through dampening increased ARC neuronal activity. PMID- 15550622 TI - United time-frequency spectroscopy for dynamics and global structure. AB - Ultrashort laser pulses have thus far been used in two distinct modes. In the time domain, the pulses have allowed probing and manipulation of dynamics on a subpicosecond time scale. More recently, phase stabilization has produced optical frequency combs with absolute frequency reference across a broad bandwidth. Here we combine these two applications in a spectroscopic study of rubidium atoms. A wide-bandwidth, phase-stabilized femtosecond laser is used to monitor the real time dynamic evolution of population transfer. Coherent pulse accumulation and quantum interference effects are observed and well modeled by theory. At the same time, the narrow linewidth of individual comb lines permits a precise and efficient determination of the global energy-level structure, providing a direct connection among the optical, terahertz, and radio-frequency domains. The mechanical action of the optical frequency comb on the atomic sample is explored and controlled, leading to precision spectroscopy with an appreciable reduction in systematic errors. PMID- 15550623 TI - Semaphorin 3E and plexin-D1 control vascular pattern independently of neuropilins. AB - The development of a patterned vasculature is essential for normal organogenesis. We found that signaling by semaphorin 3E (Sema3E) and its receptor plexin-D1 controls endothelial cell positioning and the patterning of the developing vasculature in the mouse. Sema3E is highly expressed in developing somites, where it acts as a repulsive cue for plexin-D1-expressing endothelial cells of adjacent intersomitic vessels. Sema3E-plexin-D1 signaling did not require neuropilins, which were previously presumed to be obligate Sema3 coreceptors. Moreover, genetic ablation of Sema3E or plexin-D1 but not neuropilin-mediated Sema3 signaling disrupted vascular patterning. These findings reveal an unexpected semaphorin signaling pathway and define a mechanism for controlling vascular patterning. PMID- 15550624 TI - COX-2-derived prostacyclin confers atheroprotection on female mice. AB - Female gender affords relative protection from cardiovascular disease until the menopause. We report that estrogen acts on estrogen receptor subtype alpha to up regulate the production of atheroprotective prostacyclin, PGI2, by activation of cyclooxygenase 2 (COX-2). This mechanism restrained both oxidant stress and platelet activation that contribute to atherogenesis in female mice. Deletion of the PGI2 receptor removed the atheroprotective effect of estrogen in ovariectomized female mice. This suggests that chronic treatment of patients with selective inhibitors of COX-2 could undermine protection from cardiovascular disease in premenopausal females. PMID- 15550626 TI - Science in the South. PMID- 15550627 TI - Infectious diseases. WHO gives a cautious green light to smallpox experiments. PMID- 15550628 TI - Italy. Academics protest plan to end tenure. PMID- 15550625 TI - No transcription-translation feedback in circadian rhythm of KaiC phosphorylation. AB - An autoregulatory transcription-translation feedback loop is thought to be essential in generating circadian rhythms in any model organism. In the cyanobacterium Synechococcus elongatus, the essential clock protein KaiC is proposed to form this type of transcriptional negative feedback. Nevertheless, we demonstrate here temperature-compensated, robust circadian cycling of KaiC phosphorylation even without kaiBC messenger RNA accumulation under continuous dark conditions. This rhythm persisted in the presence of a transcription or translation inhibitor. Moreover, kinetic profiles in the ratio of KaiC autophosphorylation-dephosphorylation were also temperature compensated in vitro. Thus, the cyanobacterial clock can keep time independent of de novo transcription and translation processes. PMID- 15550629 TI - Fusion energy. Euro meeting holds key to ITER Project. PMID- 15550630 TI - Paleontology. Spanish fossil sheds new light on the oldest great apes. PMID- 15550631 TI - Tobacco wars. Research on secondhand smoke questioned. PMID- 15550632 TI - Materials science. Key to cheaper, better nanotubes comes out in the wash. PMID- 15550633 TI - NIH ethics. Staff scientists protest plan to ban outside fees. PMID- 15550634 TI - AIDS vaccines. The first shot in a highly targeted strategy. PMID- 15550635 TI - Medicine. Estrogen's ties to COX-2 may explain heart disease gender gap. PMID- 15550637 TI - High-energy physics. Rara avis or statistical mirage? Pentaquark remains at large. PMID- 15550636 TI - Volcanology. Iceland's doomsday scenario? PMID- 15550638 TI - Human evolution. Faster than a hyena? Running may make humans special. PMID- 15550639 TI - American Society of Human Genetics meeting. Of worms, mice, and very old men and women. PMID- 15550640 TI - American Society of Human Genetics meeting. Ural farmers got milk gene first? PMID- 15550641 TI - American Society of Human Genetics meeting. New prostate cancer genetic link. PMID- 15550642 TI - Assisting, but not dictating. PMID- 15550643 TI - Varshavsky's contributions. PMID- 15550644 TI - Comment on "Small bilaterian fossils from 40 to 55 million years before the Cambrian". PMID- 15550646 TI - Intellectual property. Plants and intellectual property: an international appraisal. PMID- 15550647 TI - Planetary science. Alien weather at the poles of Mars. PMID- 15550648 TI - Molecular biology. Knives, accomplices, and RNA. PMID- 15550649 TI - Materials science. Shaping crystals with biomolecules. PMID- 15550650 TI - Planetary science: How Neptune pushed the boundaries of our Solar System. PMID- 15550651 TI - Structural biology. Voltage sensor meets lipid membrane. PMID- 15550653 TI - Measurement and the single particle. PMID- 15550654 TI - Getting the measure of nanotechnology. PMID- 15550655 TI - Time's romance of the decimal point. PMID- 15550656 TI - Putting the stars in their places. PMID- 15550657 TI - In the blink of an eye. PMID- 15550658 TI - Metrology and the state: science, revenue, and commerce. AB - "Natural measures of quantity, such as fathoms, cubits, inches, taken from the proportion of the human body, were once in use with every nation," taught Adam Smith in his lecture "Money as the measure of value and medium of exchange," delivered in 1763. "But by a little observation," he continued, "they found that one man's arm was longer or shorter than another's, and that one was not to be compared with the other; and therefore wise men who attended to these things would endeavour to fix upon some more accurate measure, that equal quantities might be of equal values. Their method became absolutely necessary when people came to deal in many commodities, and in great quantities of them." Smith's comments and the rationale underpinning them became increasingly urgent toward the end of the eighteenth century. PMID- 15550659 TI - Standards of time and frequency at the outset of the 21st century. AB - After 50 years of development, microwave atomic clocks based on cesium have achieved fractional uncertainties below 1 part in 10(15), a level unequaled in all of metrology. The past 5 years have seen the accelerated development of optical atomic clocks, which may enable even greater improvements in timekeeping. Time and frequency standards with various levels of performance are ubiquitous in our society, with applications in many technological fields as well as in the continued exploration of the frontiers of basic science. We review state-of-the art atomic time and frequency standards and discuss some of their uses in science and technology. PMID- 15550660 TI - The route to atomic and quantum standards. AB - Over the past half-century, there has been a shift away from standards based on particular artifacts toward those based on physical effects, the most stable being based on quantum properties of systems. This change was proposed at the end of the 19th century but is still not complete at the start of the 21st. We discuss how this vision has been implemented through recent advances in science and metrology and how these may soon lead to an SI system finally free from artifact standards, with a consistency based on fundamental constants. PMID- 15550661 TI - Quantum-enhanced measurements: beating the standard quantum limit. AB - Quantum mechanics, through the Heisenberg uncertainty principle, imposes limits on the precision of measurement. Conventional measurement techniques typically fail to reach these limits. Conventional bounds to the precision of measurements such as the shot noise limit or the standard quantum limit are not as fundamental as the Heisenberg limits and can be beaten using quantum strategies that employ "quantum tricks" such as squeezing and entanglement. PMID- 15550662 TI - The ocean takes a deep breath. AB - Deep convection is the major mechanism for replenishing oxygen in the deep interior of the world ocean, and its variability affects the use of atmospheric oxygen to monitor the global carbon cycle. Sensors mounted on autonomous floats allow this episodic breathing of the ocean to be monitored in near real time. The results suggest that the tools are available now to make oxygen a key parameter in marine global change research. PMID- 15550663 TI - Pierolapithecus catalaunicus, a new Middle Miocene great ape from Spain. AB - We describe a partial skeleton with facial cranium of Pierolapithecus catalaunicus gen. et sp. nov., a new Middle Miocene (12.5 to 13 million years ago) ape from Barranc de Can Vila 1 (Barcelona, Spain). It is the first known individual of this age that combines well-preserved cranial, dental, and postcranial material. The thorax, lumbar region, and wrist provide evidence of modern ape-like orthograde body design, and the facial morphology includes the basic derived great ape features. The new skeleton reveals that early great apes retained primitive monkeylike characters associated with a derived body structure that permits upright postures of the trunk. Pierolapithecus, hence, does not fit the theoretical model that predicts that all characters shared by extant great apes were present in their last common ancestor, but instead points to a large amount of homoplasy in ape evolution. The overall pattern suggests that Pierolapithecus is probably close to the last common ancestor of great apes and humans. PMID- 15550664 TI - Magnetic response of metamaterials at 100 terahertz. AB - An array of single nonmagnetic metallic split rings can be used to implement a magnetic resonance, which arises from an inductor-capacitor circuit (LC) resonance, at 100-terahertz frequency. The excitation of the LC resonance in the normal-incidence geometry used in our experiments occurs through the coupling of the electric field of the incident light to the capacitance. The measured optical spectra of the nanofabricated gold structures come very close to the theoretical expectations. Additional numerical simulations show that our structures exhibit a frequency range with negative permeability for a beam configuration in which the magnetic field couples to the LC resonance. Together with an electric response that has negative permittivity, this can lead to materials with a negative index of refraction. PMID- 15550665 TI - A chiral route to negative refraction. AB - Negative refraction is currently achieved by driving the magnetic permeability and electrical permittivity simultaneously negative, thus requiring two separate resonances in the refracting material. The introduction of a single chiral resonance leads to negative refraction of one polarization, resulting in improved and simplified designs of negatively refracting materials and opening previously unknown avenues of investigation in this fast-growing subject. PMID- 15550666 TI - Hertz-level measurement of the optical clock frequency in a single 88Sr+ ion. AB - The frequency of the 5s 2S(1/2)-4d 2D(5/2) electric quadrupole clock transition in a single, trapped, laser-cooled 88Sr+ ion has been measured by using an optical frequency comb referenced to a cesium fountain primary frequency standard. The frequency of the transition is measured as 444,779,044,095,484.6 (1.5) hertz, with a fractional uncertainty within a factor of 3 of that of the cesium standard. Improvements required to obtain a cesium-limited frequency measurement are described and are expected to lead to a 88Sr+ optical clock with stability and reproducibility exceeding that of the primary cesium standard. PMID- 15550667 TI - Multifunctional carbon nanotube yarns by downsizing an ancient technology. AB - By introducing twist during spinning of multiwalled carbon nanotubes from nanotube forests to make multi-ply, torque-stabilized yarns, we achieve yarn strengths greater than 460 megapascals. These yarns deform hysteretically over large strain ranges, reversibly providing up to 48% energy damping, and are nearly as tough as fibers used for bulletproof vests. Unlike ordinary fibers and yarns, these nanotube yarns are not degraded in strength by overhand knotting. They also retain their strength and flexibility after heating in air at 450 degrees C for an hour or when immersed in liquid nitrogen. High creep resistance and high electrical conductivity are observed and are retained after polymer infiltration, which substantially increases yarn strength. PMID- 15550668 TI - Water-assisted highly efficient synthesis of impurity-free single-walled carbon nanotubes. AB - We demonstrate the efficient chemical vapor deposition synthesis of single-walled carbon nanotubes where the activity and lifetime of the catalysts are enhanced by water. Water-stimulated enhanced catalytic activity results in massive growth of superdense and vertically aligned nanotube forests with heights up to 2.5 millimeters that can be easily separated from the catalysts, providing nanotube material with carbon purity above 99.98%. Moreover, patterned, highly organized intrinsic nanotube structures were successfully fabricated. The water-assisted synthesis method addresses many critical problems that currently plague carbon nanotube synthesis. PMID- 15550669 TI - Very low gene duplication rate in the yeast genome. AB - The gene duplication rate in the yeast genome is estimated without assuming the molecular clock model to be approximately 0.01 to 0.06 per gene per billion years; this rate is two orders of magnitude lower than a previous estimate based on the molecular clock model. This difference is explained by extensive concerted evolution via gene conversion between duplicated genes, which violates the assumption of the molecular clock in the analyses of duplicated genes. The average length of the period of concerted evolution and the gene conversion rate are estimated to be approximately 25 million years and approximately 28 times the mutation rate, respectively. PMID- 15550670 TI - Regulated fast nucleocytoplasmic shuttling observed by reversible protein highlighting. AB - The observation of the regulation of fast protein dynamics in a cellular context requires the development of reliable technologies. Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting. Light-induced conversion between the bright and dark states of a monomeric fluorescent protein engineered from a novel coral protein was employed. Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules. PMID- 15550671 TI - TRPM4 regulates calcium oscillations after T cell activation. AB - TRPM4 has recently been described as a calcium-activated nonselective (CAN) cation channel that mediates membrane depolarization. However, the functional importance of TRPM4 in the context of calcium (Ca2+) signaling and its effect on cellular responses are not known. Here, the molecular inhibition of endogenous TRPM4 in T cells was shown to suppress TRPM4 currents, with a profound influence on receptor-mediated Ca2+ mobilization. Agonist-mediated oscillations in intracellular Ca2+ concentration ([Ca2+]i), which are driven by store-operated Ca2+ influx, were transformed into a sustained elevation in [Ca2+]i. This increase in Ca2+ influx enhanced interleukin-2 production. Thus, TRPM4-mediated depolarization modulates Ca2+ oscillations, with downstream effects on cytokine production in T lymphocytes. PMID- 15550672 TI - A protein sensor for siRNA asymmetry. AB - To act as guides in the RNA interference (RNAi) pathway, small interfering RNAs (siRNAs) must be unwound into their component strands, then assembled with proteins to form the RNA-induced silencing complex (RISC), which catalyzes target messenger RNA cleavage. Thermodynamic differences in the base-pairing stabilities of the 5' ends of the two approximately 21-nucleotide siRNA strands determine which siRNA strand is assembled into the RISC. We show that in Drosophila, the orientation of the Dicer-2/R2D2 protein heterodimer on the siRNA duplex determines which siRNA strand associates with the core RISC protein Argonaute 2. R2D2 binds the siRNA end with the greatest double-stranded character, thereby orienting the heterodimer on the siRNA duplex. Strong R2D2 binding requires a 5' phosphate on the siRNA strand that is excluded from the RISC. Thus, R2D2 is both a protein sensor for siRNA thermodynamic asymmetry and a licensing factor for entry of authentic siRNAs into the RNAi pathway. PMID- 15550673 TI - The human polyomavirus, JCV, uses serotonin receptors to infect cells. AB - The human polyomavirus, JCV, causes the fatal demyelinating disease progressive multifocal leukoencephalopathy in immunocompromised patients. We found that the serotonergic receptor 5HT2AR could act as the cellular receptor for JCV on human glial cells. The 5HT2A receptor antagonists inhibited JCV infection, and monoclonal antibodies directed at 5HT2A receptors blocked infection of glial cells by JCV, but not by SV40. Transfection of 5HT2A receptor-negative HeLa cells with a 5HT2A receptor rescued virus infection, and this infection was blocked by antibody to the 5HT2A receptor. A tagged 5HT2A receptor colocalized with labeled JCV in an endosomal compartment following internalization. Serotonin receptor antagonists may thus be useful in the treatment of progressive multifocal leukoencephalopathy. PMID- 15550674 TI - Fat mobilization in adipose tissue is promoted by adipose triglyceride lipase. AB - Mobilization of fatty acids from triglyceride stores in adipose tissue requires lipolytic enzymes. Dysfunctional lipolysis affects energy homeostasis and may contribute to the pathogenesis of obesity and insulin resistance. Until now, hormone-sensitive lipase (HSL) was the only enzyme known to hydrolyze triglycerides in mammalian adipose tissue. Here, we report that a second enzyme, adipose triglyceride lipase (ATGL), catalyzes the initial step in triglyceride hydrolysis. It is interesting that ATGL contains a "patatin domain" common to plant acyl-hydrolases. ATGL is highly expressed in adipose tissue of mice and humans. It exhibits high substrate specificity for triacylglycerol and is associated with lipid droplets. Inhibition of ATGL markedly decreases total adipose acyl-hydrolase activity. Thus, ATGL and HSL coordinately catabolize stored triglycerides in adipose tissue of mammals. PMID- 15550675 TI - In vivo mechanisms of tissue-selective drug toxicity: effects of liver-specific knockout of the NADPH-cytochrome P450 reductase gene on acetaminophen toxicity in kidney, lung, and nasal mucosa. AB - Acetaminophen overdose causes toxicity in liver and extrahepatic tissues. Although it is well established that cytochrome P450 enzymes play a critical role in the metabolic activation of acetaminophen, it is not yet clear whether acetaminophen toxicity in extrahepatic tissues is a consequence of hepatic biotransformation. The aim of this study was to determine whether extrahepatic acetaminophen toxicity is altered in a mouse model that has liver-specific deletion of the NADPH-cytochrome P450 reductase (Cpr) gene. Liver-specific Cpr null (Null) mice were resistant to acetaminophen hepatotoxicity, and they showed faster acetaminophen clearance than did wild-type mice at a toxic acetaminophen dose (400 mg/kg i.p.). However, when circulating acetaminophen levels were made equivalent in the two strains, the severity of extrahepatic acetaminophen toxicity was decreased in the Null relative to that in the wild-type mice in the lung, kidney, and lateral nasal glands, although not in the nasal olfactory and respiratory mucosa. In the lung and liver, the decreased acetaminophen toxicity was accompanied by substantial decreases in the formation of acetaminophen protein adducts in the Null mice; adducts were not detected in other tissues examined. These results indicate that acetaminophen toxicity in the nasal mucosa is not dependent on hepatic microsomal P450-catalyzed metabolic activation and that acetaminophen toxicity in the lung, kidney, and lateral nasal glands is at least partly caused by liver-derived acetaminophen metabolites. PMID- 15550676 TI - Phosphorylation of Cytidine, Deoxycytidine, and Their Analog Monophosphates by Human UMP/CMP Kinase Is Differentially Regulated by ATP and Magnesium. AB - Human UMP/CMP kinase (cytidylate kinase; EC 2.7.4.14) is responsible for phosphorylation of CMP, UMP, and deoxycytidine monophosphate (dCMP) and also plays an important role in the activation of pyrimidine analogs, some of which are clinically useful anticancer or antiviral drugs. Previous kinetic data using recombinant or highly purified human UMP/CMP kinase showed that dCMP, as well as pyrimidine analog monophosphates, were much poorer substrates than CMP or UMP for this enzyme. This implies that other unidentified mechanisms must be involved to make phosphorylation of dCMP or pyrimidine analog monophosphates inside cells by this enzyme possible. Here, we reevaluated the optimal reaction conditions for human recombinant human UMP/CMP kinase to phosphorylate dCMP and CMP (referred as dCMPK and CMPK activities). We found that ATP and magnesium were important regulators of the kinase activities of this enzyme. Free magnesium enhanced dCMPK activity but inhibited CMPK activity. Free ATP or excess ATP/magnesium, on the other hand, inhibited dCMPK but not CMPK reactions. The differential regulation of dCMPK versus CMPK activities by ATP or magnesium was also seen in other 2' deoxypyrimidine analog monophosphates (deoxyuridine monophosphate, 5 fluorodeoxyuridine monophosphate, 1-beta-D-arabinofuranosylcytosine monophosphate, and gemcitabine monophosphate) versus their ribose-counterparts (UMP and 5-fluorouridine monophosphate), in a similar manner. The data suggest that the active sites of human UMP/CMP kinase for dCMP and for CMP cannot be identical. Furthermore, enzyme inhibition studies demonstrated that CMP could inhibit dCMP phosphorylation in a noncompetitive manner, with Ki values much higher than its own Km values. We thus propose novel models for the phosphorylation action of human UMP/CMP kinase. PMID- 15550677 TI - Regulation of serum response factor-dependent gene expression by proteasome inhibitors. AB - Serum response factor (SRF) is activated by contractile and hypertrophic agonists, such as endothelin-1 (ET1) to stimulate expression of cytoskeletal proteins in vascular smooth muscle cells (VSMCs). While studying the regulation of smooth muscle alpha-actin (SMA) expression at the level of protein stability, we discovered that inhibition of proteasome-dependent protein degradation by N benzoyloxycarbonyl (Z)-Leu-Leu-leucinal (MG132) or lactacystin (LC) did not enhance the levels of SMA, but, unexpectedly, attenuated SMA expression in response to ET1, without affecting the viability of VSMCs. Down-regulation of SMA protein by MG132 or LC occurred at the level of SMA transcription and via the inhibition of SRF activity. By contrast, MG132 and LC potentiated the activity of activator protein-1 transcription factor. Regulation of SRF by MG132 was not related to inhibition of nuclear factor-kappaB, an established target of proteasome inhibitors, and was not mediated by protein kinase A, a powerful regulator of SRF activity. Signaling studies indicate that inhibition of ET1 induced SRF activity by MG132 occurs at the level downstream of heterotrimeric G proteins Gq/11 and G13, of small GTPase RhoA, and of actin dynamics but at the level of SRF-DNA binding. MG132 treatment did not result in ubiquitination or accumulation of SRF. By contrast, the levels of c-Jun were rapidly increased upon incubation of cells with MG132, and ectopic overexpression of c-Jun mimicked the effect of MG132 on SRF activity. Together, these data suggest that inhibition of proteasome results in down-regulation of SMA expression via up-regulation of c Jun and repression of SRF activity at the level of DNA binding. PMID- 15550678 TI - Activation of the melastatin-related cation channel TRPM3 by D-erythro sphingosine [corrected]. AB - TRPM3, a member of the melastatin-like transient receptor potential channel subfamily (TRPM), is predominantly expressed in human kidney and brain. TRPM3 mediates spontaneous Ca2+ entry and nonselective cation currents in transiently transfected human embryonic kidney 293 cells. Using measurements with the Ca2+ sensitive fluorescent dye fura-2 and the whole-cell patch-clamp technique, we found that D-erythro-sphingosine, a metabolite arising during the de novo synthesis of cellular sphingolipids, activated TRPM3. Other transient receptor potential (TRP) channels tested [classic or canonical TRP (TRPC3, TRPC4, TRPC5), vanilloid-like TRP (TRPV4, TRPV5, TRPV6), and melastatin-like TRP (TRPM2)] did not significantly respond to application of sphingosine. Sphingosine-induced TRPM3 activation was not mediated by inhibition of protein kinase C, depletion of intracellular Ca2+ stores, and intracellular conversion of sphingosine to sphingosine-1-phosphate. Although sphingosine-1-phosphate and ceramides had no effect, two structural analogs of sphingosine, dihydro-D-erythro-sphingosine and N,N-dimethyl-D-erythro-sphingosine, also activated TRPM3. Sphingolipids, including sphingosine, are known to have inhibitory effects on a variety of ion channels. Thus, TRPM3 is the first ion channel activated by sphingolipids. PMID- 15550679 TI - Identification of endogenous glucocorticoid repressed genes differentially regulated by a glucocorticoid receptor mutant able to separate between nuclear factor-kappaB and activator protein-1 repression. AB - Glucocorticoids are commonly used in the clinic, but long-term treatment is often associated with severe side effects. One way to reduce unwanted effects is to restrict glucocorticoid receptor (GR) signaling through defined pathways. In this study, we examine endogenous target genes regulated by a GR mutant that in contrast to the wild-type GR is unable to repress stimulated nuclear factor kappaB (NF-kappaB) activity, whereas repression of activator protein-1 (AP-1) activity is maintained. This GR mutant (GRR488Q) harbors a point mutation in the second zinc finger of the DNA binding domain. Its ability to distinguish between NF-kappaB and AP-1 repression is defined using reporter genes regulated by both simple and natural promoters. The inability of GRR488Q to repress NF-kappaB was not related to its inability to activate target genes through a glucocorticoid response element. Furthermore, the discriminating property was observed in three different cell lines, suggesting that this is not a cell-specific effect. These results show that different receptor surfaces or mechanisms are involved in repression of NF-kappaB and AP-1, respectively. It is interesting that the GRR488Q still interacted physically with NF-kappaB. Gene expression profiling of human embryonic kidney 293 cells, which express the wild-type GR and the GRR488Q mutant allowed identification of endogenous genes preferentially repressed by GR interference with NF-kappaB activity. The genes differentially regulated by GRR488Q mutant versus the wild-type GR after 2 h of treatment seem mainly to be involved in control of transcription and cell growth. At 8 h, no such distinction could be seen. PMID- 15550680 TI - The aryl hydrocarbon receptor predisposes hepatocytes to Fas-mediated apoptosis. AB - Liver homeostasis is achieved by the removal of diseased and damaged hepatocytes and their coordinated replacement to maintain a constant liver cell mass. Cirrhosis, viral hepatitis, and toxic drug effects can all trigger apoptosis in the liver as a means of removing the unwanted cells, and the Fas "death receptor" pathway comprises a major physiological mechanism by which this occurs. The susceptibility to Fas-mediated apoptosis is, in part, a function of the hepatocyte's proteome. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor known to influence apoptosis, conceivably by regulating the expression of genes involved in apoptotic signaling. In this article, we present evidence demonstrating that AhR expression and function promote apoptosis in liver cells in response to Fas stimulation. Reintroduction of the AhR into the AhR-negative BP8 hepatoma cells as well as into primary hepatocytes from AhR knockout mice increases the magnitude of cell death in response to Fas ligand. Enhanced apoptosis correlates with increased caspase activity and mitochondrial cytochrome c release but not with the expression of several Bcl-2 family proteins. In vivo studies showed that in contrast to wild-type mice, AhR knockout mice are protected from the lethal effects of the anti-Fas Jo2 antibody. Moreover, down-regulation of the aryl hydrocarbon receptor nuclear translocator protein in vivo by adenovirus-mediated RNA interference to suppress AhR activity provided wild-type mice partial protection from Jo2-induced lethality. PMID- 15550682 TI - Protease inhibitors in spontaneous cervical artery dissections. AB - BACKGROUND AND PURPOSE: Observations in patients with arterial aneurysms, fibromuscular dysplasia, and spontaneous cervical artery dissection (sCAD) indicate that protease inhibitor deficiency might boost the enzymatic destruction of arterial tissue and increase the risk of these arterial wall diseases. Here we present the first large investigation of the protease inhibitor hypothesis in patients with sCAD. METHODS: Eighty patients with sCAD were compared with 80 age- and sex-matched healthy individuals. Alpha1-antitrypsin (alpha1-AT) and alpha2 macroglobulin (alpha2-MG) levels, and alpha1-AT genotypes were assessed and compared between groups. RESULTS: alpha1-AT and alpha2-MG levels as well as alpha1-AT genotypes did not differ significantly between patients and controls. The frequency of Z alleles in the patient group was higher than in the control group and than in other cohorts from Europe; however, the difference remained nonsignificant. All patients with Z alleles had internal carotid artery dissections. CONCLUSIONS: Overall, this data does not support the hypothesis that protease inhibitor levels or alpha1-AT genotypes play an important role in the etiology of sCAD. The present data does not exclude that the Pi-Z allele might have an influence on subgroups of sCAD, such as internal carotid artery dissections. PMID- 15550686 TI - Angiogenesis in symptomatic intracranial atherosclerosis: predominance of the inhibitor endostatin is related to a greater extent and risk of recurrence. AB - BACKGROUND AND PURPOSE: Angiogenesis may be beneficial in chronic myocardial and limb ischemia, but its role in intracranial atherosclerosis remains unknown. We aimed to investigate the relationship between the pro-angiogenic vascular endothelial growth factor (VEGF) and the anti-angiogenic endostatin, and the extent and risk of recurrence of symptomatic intracranial atherosclerosis. METHODS: Of a total of 94 consecutive patients with symptomatic intracranial stenoses, 40 fulfilled all inclusion criteria. Intracranial stenoses were confirmed by magnetic resonance angiography. Magnetic resonance imaging (MRI) including diffusion-weighted sequences was conducted. Plasmatic VEGF and endostatin were determined from blood samples obtained 3 months after stroke onset, and patients were followed-up thereafter. RESULTS: A total of 144 intracranial stenoses were confirmed (median number per patient=3). Endostatin/VEGF ratio gradually augmented with the increasing number of intracranial stenoses (r=0.35, P=0.02). Diabetes mellitus (OR, 6.04; CI, 1.1 to 32.2; P=0.03) and a higher endostatin/VEGF ratio (OR, 15.7; CI, 2.2 to 112.3; P=0.006) were independently associated with a greater extent of intracranial atherosclerosis. During a median follow-up of 13 months, 8 patients (20%) experienced a new cerebral ischemic event. A higher baseline endostatin concentration was an independent predictor of new events (hazard ratio, 7.24; CI, 1.6 to 33.8; P=0.011) in a Cox regression model after adjustment for age, sex, number of stenotic vessels, and risk factors. Patients with a higher endostatin level had a lower survival free of new events (P=0.01, log-rank test). CONCLUSIONS: A predominance of the inhibitor endostatin within the endogenous angiogenic response is associated with a greater extent and risk of recurrence of symptomatic intracranial atherosclerosis, suggesting that angiogenesis may be beneficial in this condition. PMID- 15550687 TI - Molecular signatures of vascular injury are associated with early growth of intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: To investigate whether molecular markers of inflammation and endothelial injury are associated with early growth of intracerebral hemorrhage (ICH). METHODS: In a multicenter prospective study, we determined concentrations of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), matrix metalloproteinase-9 (MMP-9), and cellular fibronectin (c-Fn) in blood samples obtained on admission from 183 patients with primary hemispheric ICH of <12 hours' duration. Patients had a neurological evaluation and a computed tomography (CT) scan performed at baseline and at 48+/-6 hours. Early growth of the ICH was defined as a volume increase >33% between the 2 CT examinations for ICH with a baseline volume <20 mL and >10% for ICH > or =20 mL. Clinical, radiological, and biochemical predictive factors of ICH enlargement were analyzed by logistic regression analysis. RESULTS: Fifty-four (29.5%) patients showed a relevant early growth of ICH. High leukocyte count and fibrinogen levels, low platelet count, and intraventricular bleeding were associated with early ICH growth in bivariate analyses. Plasma concentrations of IL-6 (median [quartiles]: 19.6 [13.6; 29.9] versus 15.9 [11.5; 19.8] pg/mL), TNF-alpha (13.5 [8.4; 30.5] versus 8.7 [4.7; 13.5] pg/mL), MMP-9 (153.3 [117.7; 204.7] versus 70.6 [47.8; 103.8] ng/mL), and c-Fn (8.8 [6.2; 12.5] versus 2.8 [1.6; 4.2] microg/mL) were significantly higher in patients with early growth of ICH (all P<0.001). C-Fn levels >6 microg/mL (OR, 92; 95%CI, 22 to 381; P<0.0001) and IL-6>24 pg/mL (OR, 16; 95%CI, 2.3 to 119; P=0.005) were independently associated with ICH enlargement in the logistic regression analysis. CONCLUSIONS: Molecular signatures of vascular injury and inflammatory markers in the early acute phase of ICH are associated with subsequent enlargement of the hematoma. PMID- 15550689 TI - Reconstruction of the patterns of gene expression in the developing mouse heart reveals an architectural arrangement that facilitates the understanding of atrial malformations and arrhythmias. AB - Firm knowledge about the formation of the atrial components and of the variations seen in congenital cardiac malformations and abnormal atrial rhythms is fundamental to our understanding of the normal structure of the definitive atrial chambers. The atrial region is relatively inaccessible and has continued to be the source of disagreement. Seeking to resolve these controversies, we made three dimensional reconstructions of the myocardial components of the developing atrium, identifying domains on the basis of differential expression of myocardial markers, connexin40, and natriuretic precursor peptide A. These reconstructions, made from serial sections of mouse embryos, show that from the outset of atrial development, the systemic and pulmonary veins are directly connected to the atrium. Relative to the systemic junctions, however, the pulmonary venous junction appears later. Our experience shows that three-dimensional reconstructions have three advantages. First, they provide clear access to the combined morphological and molecular data, allowing clarification and verification of morphogenetic concepts for nonmorphological experts and setting the scene for further discussion. Second, they demonstrate that, from the outset, the myocardium surrounding the pulmonary veins is distinct from that clothing the systemic venoatrial junctions. Third, they reveal an anatomical and molecular continuity between the entrance of the systemic venous tributaries, the internodal atrial myocardium, and the atrioventricular region. All these regions are derived from primary myocardium, providing a molecular basis for the observed nonrandom distribution of focal right atrial tachycardias. PMID- 15550690 TI - Cardiac sodium-calcium exchanger is regulated by allosteric calcium and exchanger inhibitory peptide at distinct sites. AB - The sarcolemmal Na+-Ca2+ exchanger (NCX) is the main Ca2+ extrusion mechanism in cardiac myocytes and is thus essential for the regulation of Ca2+ homeostasis and contractile function. A cytosolic region (f-loop) of the protein mediates regulation of NCX function by intracellular factors including inhibition by exchanger inhibitory peptide (XIP), a 20 amino acid peptide matching the sequence of an autoinhibitory region involved in allosteric regulation of NCX by intracellular Na+, Ca2+, and phosphatidylinositol-4,5-biphosphate (PIP2). Previous evidence indicates that the XIP interaction domain can be eliminated by large deletions of the f-loop that also remove activation of NCX by intracellular Ca2+. By whole-cell voltage clamping experiments, we demonstrate that deletion of residues 562-679, but not 440- 456, 498-510, or 680-685 of the f-loop selectively eliminates XIP-mediated inhibition of NCX expressed either heterologously (HEK293 and A549 cells) or in guinea pig cardiac myocytes. In contrast, by plotting I(NCX) against reverse-mode NCX-mediated Ca2+ transients in myocytes, we demonstrate that Ca2+-dependent regulation of NCX is preserved in Delta562-679, but significantly reduced in the other three deletion mutants. The findings indicate that f-loop residues 562-679 may contain the regulatory site for endogenous XIP, but this site is distinct from the Ca2+-regulatory domains of the NCX. Because regulation of the NCX by Na+ and PIP2 involves the endogenous XIP region, the Delta562-679 mutant NCX may be a useful tool to investigate this regulation in the context of the whole cardiac myocyte. PMID- 15550691 TI - Small artery remodeling depends on tissue-type transglutaminase. AB - Remodeling of small arteries is essential in the long-term regulation of blood pressure and blood flow to specific organs or tissues. A large part of the change in vessel diameter may occur through non-growth-related reorganization of vessel wall components. The hypothesis was tested that tissue-type transglutaminase (tTG), a cross-linking enzyme, contributes to the inward remodeling of small arteries. The in vivo inward remodeling of rat mesenteric arteries, induced by low blood flow, was attenuated by inhibition of tTG. Rat skeletal muscle arteries expressed tTG, as identified by Western blot and immunostaining. In vitro, activation of these arteries with endothelin-1 resulted in inward remodeling, which was blocked by tTG inhibitors. Small arteries obtained from rats and pigs both showed inward remodeling after exposure to exogenous transglutaminase, which was inhibited by addition of a nitric oxide donor. Enhanced expression of tTG, induced by retinoic acid, increased inward remodeling of porcine coronary arteries kept in organ culture for 3 days. The activity of tTG was dependent on pressure. Inhibition of tTG reversed remodeling, causing a substantial increase in vessel diameter. In a collagen gel contraction assay, tTG determined the compaction of collagen by smooth muscle cells. Collectively, these data show that small artery remodeling associated with chronic vasoconstriction depends on tissue-type transglutaminase. This mechanism may reveal a novel therapeutic target for pathologies associated with inward remodeling of the resistance arteries. PMID- 15550693 TI - Mouse strain-specific differences in vascular wall gene expression and their relationship to vascular disease. AB - OBJECTIVE: Different strains of inbred mice exhibit different susceptibility to the development of atherosclerosis. The C3H/HeJ and C57Bl/6 mice have been used in several studies aimed at understanding the genetic basis of atherosclerosis. Under controlled environmental conditions, variations in susceptibility to atherosclerosis reflect differences in genetic makeup, and these differences must be reflected in gene expression patterns that are temporally related to the development of disease. In this study, we sought to identify the genetic pathways that are differentially activated in the aortas of these mice. METHODS AND RESULTS: We performed genome-wide transcriptional profiling of aortas from C3H/HeJ and C57Bl/6 mice. Differences in gene expression were identified at baseline as well as during normal aging and longitudinal exposure to high-fat diet. The significance of these genes to the development of atherosclerosis was evaluated by observing their temporal pattern of expression in the well-studied apolipoprotein E model of atherosclerosis. CONCLUSIONS: Gene expression differences between the 2 strains suggest that aortas of C57Bl/6 mice have a higher genetic propensity to develop inflammation in response to appropriate atherogenic stimuli. This study expands the repertoire of factors in known disease-related signaling pathways and identifies novel candidate genes for future study. To gain insights into the molecular pathways that are differentially activated in strains of mice with varied susceptibility to atherosclerosis, we performed comprehensive transcriptional profiling of their vascular wall. Genes identified through these studies expand the repertoire of factors in disease-related signaling pathways and identify novel candidate genes in atherosclerosis. PMID- 15550692 TI - Cells expressing early cardiac markers reside in the bone marrow and are mobilized into the peripheral blood after myocardial infarction. AB - The concept that bone marrow (BM)-derived cells participate in cardiac regeneration remains highly controversial and the identity of the specific cell type(s) involved remains unknown. In this study, we report that the postnatal BM contains a mobile pool of cells that express early cardiac lineage markers (Nkx2.5/Csx, GATA-4, and MEF2C). These cells are present in significant amounts in BM harvested from young mice but their abundance decreases with age; in addition, the responsiveness of these cells to gradients of motomorphogens SDF-1, HGF, and LIF changes with age. FACS analysis, combined with analysis of early cardiac markers at the mRNA and protein levels, revealed that cells expressing these markers reside in the nonadherent, nonhematopoietic CXCR4+/Sca-1+/lin-/CD45 mononuclear cell (MNC) fraction in mice and in the CXCR4+/CD34+/AC133+/CD45- BMMNC fraction in humans. These cells are mobilized into the peripheral blood after myocardial infarction and chemoattracted to the infarcted myocardium in an SDF-1-CXCR4-, HGF-c-Met-, and LIF-LIF-R-dependent manner. To our knowledge, this is the first demonstration that the postnatal BM harbors a nonhematopoietic population of cells that express markers for cardiac differentiation. We propose that these potential cardiac progenitors may account for the myocardial regenerative effects of BM. The present findings provide a novel paradigm that could reconcile current controversies and a rationale for investigating the use of BM-derived cardiac progenitors for myocardial regeneration. PMID- 15550694 TI - Asymptomatic carotid atherosclerosis is associated with circulating chlamydia pneumoniae DNA in younger normotensive subjects in a general population survey. AB - OBJECTIVE: Chlamydia pneumoniae infection has been associated with atherosclerosis, but serodiagnosis is unreliable in predicting vascular infection. Direct detection of circulating chlamydial DNA in peripheral blood mononuclear cells (PBMCs) was thus evaluated as a marker for cardiovascular risk in a general population survey using the common carotid intima-media thickness (IMT) as surrogate marker of asymptomatic atherosclerosis. METHODS AND RESULTS: C pneumoniae DNA in PBMCs was determined by nested polymerase chain reaction and associated with IMT for 1032 healthy participants of a general population survey who were within the highest or lowest IMT distribution quartile. C pneumoniae DNA was more prevalent in those with increased IMT (13.4% versus 10.7%), but this was not significant in univariate and of borderline significance in multivariate analysis. Testing for potential effect modifications by known strong determinants of an increased IMT in group interaction analysis revealed an independent association between C pneumoniae DNA and IMT in normotensive subjects (odds ratio [OR], 2.06; 95% CI, 1.05 to 4.03; P=0.04) and in those <70 years old (OR, 1.84; 95% CI, 1.06 to 3.19; P=0.03). CONCLUSIONS: Asymptomatic atherosclerosis is associated with circulating C pneumoniae DNA independently of classical cardiovascular risk factors in normotensive subjects and those <70 years old. C pneumoniae has been implicated in atherogenesis. We determined the association of chlamydial DNA in peripheral blood mononuclear cells with the carotid intima media thickness from 1032 healthy subjects from a general population survey. A stratified group interaction analysis revealed an independent association in normotensive subjects and those <70 years old. PMID- 15550695 TI - Betaine and folate status as cooperative determinants of plasma homocysteine in humans. AB - OBJECTIVE: Two published studies have demonstrated that betaine in the circulation is a determinant of plasma total homocysteine, but none had sufficient power to investigate the possible effect modification by folate status. METHODS AND RESULTS: We measured homocysteine, betaine, folate, vitamin B(6), and related compounds in serum/plasma from 500 healthy men and women aged 34 to 69 years before (fasting levels) and 6 hours after a standard methionine loading test. Choline, dimethylglycine, and folate were determinants of plasma betaine in a multiple regression model adjusting for age and sex. The increase in homocysteine after loading showed a strong inverse association with plasma betaine and a weaker inverse association with folate and vitamin B(6). Fasting homocysteine showed a strong inverse relation to folate, a weak relation to plasma betaine, and no relation to vitamin B(6). Notably, adjusted (for age and sex) dose-response curves for the postmethionine increase in homocysteine or fasting homocysteine versus betaine showed that the inverse associations were most pronounced at low serum folate, an observation that was confirmed by analyses of interaction. CONCLUSIONS: Collectively, these results show that plasma betaine is a strong determinant of increase in homocysteine after methionine loading, particularly in subjects with low folate status. In 500 healthy subjects, postmethionine load increase in tHcy showed a stronger inverse relation to betaine than to folate and vitamin B6, whereas for fasting tHcy, betaine was a weaker determinant than folate. For both tHcy modalities, the association with betaine was most pronounced in subjects with low folate status. PMID- 15550697 TI - From Monitoring of Physical Agents to the Assessment of Environmental Exposure. Torino, Italy, 29-31 October 2003. Workshop proceedings. PMID- 15550696 TI - Coagulation factor Xa stimulates interleukin-8 release in endothelial cells and mononuclear leukocytes: implications in acute myocardial infarction. AB - OBJECTIVE: In acute myocardial infarction (AMI), proinflammatory plasma C reactive protein values are strongly associated with postinfarction morbidity and mortality. So far, the cause of these inflammatory changes is not well understood. Therefore, we sought to investigate the relationship between the activation of coagulation and subsequent systemic inflammatory changes in AMI. METHODS AND RESULTS: Factor Xa (FXa) bound to tissue factor pathway inhibitor and prothrombin fragments F1+2 (F1+2) were used as a measure for activated coagulation. To assess systemic inflammatory changes, plasma interleukin (IL)-6 and IL-8 concentrations were analyzed by immunoassay. Blood samples were taken from 21 patients with AMI and 20 patients with stable angina pectoris. In AMI, tissue factor pathway inhibitor FXa but not F1+2 plasma levels were associated with circulating IL-8 (P=0.01). In vitro experiments revealed that FXa stimulated IL-8 and monocyte chemoattractant protein-1 release and RNA expression in endothelial cells and mononuclear leukocytes by activation of protease-activated receptor-1. CONCLUSIONS: Our data suggest that coagulation FXa may contribute to proinflammatory changes in AMI by stimulation of IL-8 release. Therapeutic inhibition of the proinflammatory effects of FXa may improve the clinical course in AMI. This study investigates the relationship between the activation of coagulation and systemic inflammatory changes in acute myocardial infarction. Tissue factor pathway inhibitor factor Xa but not F1+2 plasma levels were associated with circulating interleukin-8. In vitro factor Xa stimulated interleukin-8 and monocyte chemoattractant protein-1 release and RNA expression by activation of protease-activated receptor 1 as an underlying mechanism. PMID- 15550698 TI - Ray-tracing techniques to assess the electromagnetic field radiated by radio base stations: application and experimental validation in an urban environment. AB - This paper aims to validate a ray-tracing model for electromagnetic field calculation, which is used in urban environments to predict irradiation from radio base stations for population exposure evaluation. Validation was carried out through a measurement campaign by choosing measurement points in order to test different propagation environments and analysing broadcast control channels through narrow band measurements. Comparison of the calculated and measured fields indicates that the ray-tracing model used calculates electric field with good accuracy, in spite of the fact that the propagation environment is not described in detail, because of difficulties in modelling the geometrical and electrical characteristics of urban areas. Differences between the calculated and measured results remain below 1.5 dB, with a mean value of 1 dB. PMID- 15550699 TI - Magnetic fields dispersed by high-voltage power lines: an advanced evaluation method based on 3-D models of electrical lines and the territory. AB - An advanced numerical evaluation tool is proposed for calculating the magnetic flux density dispersed by high-voltage power lines. When compared to existing software packages based on the application of standardized methods, this tool turned out to be particularly suitable for making accurate evaluations on vast portions of the territory, especially when the contribution of numerous aerial and/or underground lines must be taken into account. The aspects of the tool of greatest interest are (1) the interaction with an electronic archive of power lines, from which all the information necessary for the calculation is obtained; (2) the use of three-dimensional models of both the power lines and the territory crossed by these; (3) the direct interfacing with electronic cartography; and finally (4) the use of a representation procedure for the results that is based on contour maps. The tool had proven to be very useful especially for Environmental Impact Assessment procedures relative to new power lines. PMID- 15550700 TI - Hearing aids' electromagnetic immunity to environmental RF fields. AB - In this work, the electromagnetic interference on hearing aids was evaluated. Electromagnetic (EM) immunity tests on different types of hearing aids were carried out, using signals of intensity and modulation comparable to those present in the environment. The purpose of this work is to characterise the interference, establishing the immunity threshold for different frequencies and finding out which types of hearing aids are more susceptible, and in which frequency range. The tests were carried out in a GTEM cell on seven hearing aids, using AM and GSM signals in the radiofrequency (RF) range. PMID- 15550701 TI - Radon concentration in waters of geothermal Euganean basin--Veneto, Italy. AB - Since ancient times the warm thermal waters of Euganean basin, Italy, have been used for therapeutic purposes. The radioactive characteristics, assumed in depth, are due, in particular, to radon gas, which when released during the therapies in the indoor thermal room, determines exposure for the workers. The preliminary results of the project, the purpose of which is to analyze the totality of thermal springs, are presented. The concentrations, obtained by gamma spectrometry, show a high variability, and are comparable to those in other thermal springs in Italy and abroad. Possible correlations with geophysical factors, such as the temperature and depth of springs, have been investigated. A geostatistical analysis of the radon data has also been carried out, through the study of the experimental variogram, obtained by kriging one map of the radon concentration: such an approach is useful not only for mapping but also for giving one possible interpretation of the examined phenomenon through structural geologic characteristics. PMID- 15550702 TI - Quality assurance and quality control methodologies used within the Austrian UV monitoring network. AB - The Austrian UVB monitoring network is operational since 1997. Nine detectors for measuring erythemally weighted solar UV irradiance are distributed over Austria in order to cover the main populated areas as well as different levels of altitude. The detectors are calibrated to indicate the UV-Index, the internationally agreed unit for erythemally weighted solar UV irradiance. Calibration is carried out in the laboratory for determination of spectral sensitivity of each detector, and under the sun for absolute comparison with a well-calibrated, double-monochromator spectroradiometer. For the conversion from detector-weighted units to erythemally weighted units a lookup table is used, which is calculated using a radiative transfer model and which reflects the dependence of the conversion on the solar zenith angle and total ozone content of the atmosphere. The uncertainty of the calibration is about +/-7%, dominated by the uncertainty of the calibration lamp for the spectroradiometer (+/-4%). The long-term stability of this type of detectors has been found to be not satisfactory. Therefore, routinely every year all detectors are completely recalibrated. Variations of the calibration factors up to +/-10% are found. Thus, during routine operation, several measures take place for quality control. The measured data are compared to results of model calculations with a radiative transfer model, where clear sky and an aerosol-free atmosphere are assumed. At each site, the UV data are also compared with data from a co-located pyranometer measuring total solar irradiance. These two radiation quantities are well correlated, especially on clear days and when the ozone content is taken into account. If suspicious measurements are found for one detector in the network, a well-calibrated travelling reference detector of the same type is set up side-by side, allowing the identification of relative differences of approximately 3%. If necessary, a recalibration is carried out. As the main aim for the Austrian UV monitoring network is the information of the public about the actual levels of UVB irradiance, the measurement results are published on-line in the Internet (http://www.uibk.ac.at/projects/uv-index). For the previous day and backwards approximately 2 years, the distribution of maximal UVB levels over Austria is shown on a regional map. Additionally, near real-time data of most of the measurement stations are presented with a delay of usually less than half-an hour. Together with these actually measured data there is also shown the diurnal variation of the maximal expected value of the UV-Index under ideal clear conditions. PMID- 15550703 TI - A new method to measure the acoustic surface impedance outdoors. AB - In the European countries noise pollution is considered to be one of the most important environmental problems. With respect to traffic noise, different researchers are working on the reduction of noise at the source, on the modelling of the acoustic absorption of the road structure and on the effects of the pavement on the propagation. The aim of this paper is to propose a new method to measure the acoustic impedance of surfaces located outdoors, which allows us to further noise propagation models, in order to evaluate exactly the noise exposure. PMID- 15550704 TI - Influence of probe size on the measurement accuracy of non-uniform ELF magnetic fields. AB - The accuracy of extremely low frequency (ELF) magnetic field measurements can be sensitively varied by the meter probe size when the spatial distribution of the magnetic flux density is highly non-uniform, e.g. if the measurement point is close to the field source. The deviation between measured and actual field varies from point to point depending on the probe dimension and on the source configuration, once the other measurement conditions are fixed. The analysis of this effect is developed through a two-dimensional numerical model that enables the evaluation of the actual field value and of the corresponding probe indication. The error distribution, i.e. the deviation between actual and measured value, is computed for magnetic fields generated by industrial three phase systems under both balanced and unbalanced supply conditions. The analysis shows how, for a given source, the error depends not simply on the distance from the source in relation to the probe size, but on its spatial distribution, which is complex and cannot be a priori predicted without using a computational tool such as the one proposed. PMID- 15550705 TI - The monitoring of tap waters in Milano: planning, methods and results. AB - The radiometric monitoring of tap waters has been carried out, in Northern Italy, since late the 1980s as part of the activities headed by the National Surveillance Network on environmental radioactivity. Measurements were accomplished by gamma spectrometry over large samples of water that were drained in a single place and concentrated by ion exchange resin treatment. More recently a regular, periodical monitoring was started using liquid scintillation measurements. In this manner, gross alpha and beta activities, uranium, (226)Ra and (222)Rn concentrations are determined at present. Nevertheless, a single point sampling may be not fully representative of the whole Milano aqueduct, significantly extended and fed by a number of wells. As a control, an extended monitoring of the aqueduct was planned and variations of water radioactivity in different Milano areas have been assessed. PMID- 15550706 TI - Radiocaesium in mushrooms from northeast Italy, 1986-2002. AB - Late in the summer of 1986, the Health Physics Departments of Pordenone, Udine and Trieste, entrusted with monitoring radioactivity in the environment and food as a consequence of the Chernobyl accident, started noticing high concentrations of radionuclides--especially radiocaesium--in mushroom samples coming from different areas of the Friuli Venezia Giulia region (northeast Italy). To date, the authors have conducted 14 annual rounds of sampling and gamma spectrometry measurements on mushrooms, generating a total of over 2250 samples belonging to more than 300 species, which were picked in about 30 stations in the region. This surveys the main results from 15 years of macromycetes radiocontamination analysis in the region, the still unsolved problems, and hypotheses for future work. PMID- 15550707 TI - The percentage of the population exposed to harmful acoustic pollution levels resulting from vehicular traffic in the Hospital' area of Turin. AB - This paper presents the methodology and the main results of a study that has as its principal aim, the experimental identification the application and the quantification of the sustainable indicator 'Percentage of the population exposed to acoustic pollution levels' in the 'Hospital' area of Turin. The investigation of this indicator was prompted by the province of Turin's previous categorisation as 'Lead City' in the project 'Towards a support description at local the level- European Community Indicators'. Alongside, the process of experimenting, applying and quantifying the sustainable indicator--and as a logical fit with what was foreseen by the Italian normative regulation on protection against environmental acoustic pollution--the study supplies the base elements to analyse the environmental acoustic climate conditions in an important area of the city and defines the interaction tools that are the environmental indicators for the town council's future Acoustic Restoration Plan (PRA). PMID- 15550708 TI - Evaluation of the electromagnetic field level emitted by medium frequency AM broadcast stations. AB - In order to estimate the level of the electromagnetic field produced by telecommunication systems, different computational techniques can be employed whose complexity depends on the accuracy of the final results. In this paper, we present the validation of a code based on the method of moments that allows us to analyse the electromagnetic field emitted by radio-communication systems operating at medium frequencies. The method is able to provide an accurate estimate of the levels of electromagnetic field produced by this type of device and, consequently, it can be used as a method for verifying the compliance of the system with the safe exposure level regulations and population protection laws. Some numerical and experimental results are shown relevant to an amplitude modulated (AM) radio transmitter, together with the results of a forthcoming system that will be operative in the near future. PMID- 15550709 TI - UMTS signal measurements with digital spectrum analysers. AB - The launch of the Universal Mobile Telecommunications System (UMTS), the most recent mobile telecommunications standard has imposed the requirement of updating measurement instrumentation and methodologies. In order to define the most reliable measurement procedure, which is aimed at assessing the exposure to electromagnetic fields, modern spectrum analysers' features for correct signal characterisation has been reviewed. PMID- 15550711 TI - A European satellite-derived UV climatology available for impact studies. AB - This paper presents a satellite-derived climatology of the surface UV radiation, intended to support impact studies on the environment and human health. As of today, the dataset covers the period from 1 January 1984 to 31 August 2003, with daily dose maps covering Europe with a spatial resolution of 0.05 degrees. A comparison between the modelled erythemal daily dose and measurements in Ispra yields an r.m.s. value with a relative difference of 29% and a bias of 3%. The seemingly large dispersion is, however, due to a restricted number of days for which the relative difference is very high. The climatological dataset documents systematic patterns in the geographical distribution of the surface UV radiation due to cloudiness, altitude and snow. It also shows a large year-to-year variability in monthly doses of upto +/-50% in spring and +/-30% in summer. PMID- 15550710 TI - Determinants of exposure to electromagnetic fields from mobile phones. AB - In actual conditions of use, the power radiated from cellular phones changes during conversation depending on several factors. Upon request from the radio base station (RBS), the phone in fact, reduces, its power to a level that is deemed optimum for the quality of conversation. In this study, special phones, which had been modified to allow the continuous logging of power emitted during the calls have been used. Off-line processing of recorded data allowed the analysis of the behaviour of mobile phones under real-use conditions. Further data recorded by operators at selected base stations were used for the purposes of comparison and checking of the effectiveness of the experimental method. The results indicate a high proportion of use of the highest power levels, under any circumstance. Such behaviour is mainly due to frequent handovers requested by the control software to optimise the communication traffic. PMID- 15550713 TI - Hypothesis of acoustic characterisation of a 'life place'. AB - In this job a hypothesis of descriptive mathematical model is described that permits to characterize the destined areas to the conservation of the nature and the life places ("biotope"), to the aim to define the degree of existing problems, from the acoustic point of view. In agreement with European Directive 2002/49/CE (Attached I), the additional acoustic indications more adapted to the characterization of "silent zones in opened campain" have been characterized. Primary purpose of the present study is to recognize and to characterize single present sources of noise in a life place, distinguishing living natural sources from those unliving ones, and anthropic technological ones from those anthropic social ones, concentrating the attention on the definition of a weighting criterion for the produced acoustic impact from several elements of pressure that interact on a protected natural area. The data collection is in course; the result will serve to define the intervals of the acceptability classes. PMID- 15550712 TI - A method to evaluate the contribution of building material to indoor gamma dose rate through outdoor measurements: preliminary results. AB - Building materials can be an important source of exposure to gamma radiation indoors. In situations in which it is impossible to enter the dwellings to measure indoor gamma dose rate, as can happen in epidemiological studies and in surveys on randomly selected dwellings, it is important to obtain accurate and precise estimates of the indoor gamma dose rate. In this paper, preliminary results of a validation study of a new method, named IN-OUT, to estimate the indoor gamma dose rate attributable to the building materials are presented. This method, which is still in progress, is based on outdoor measurements, performed close to an external wall of a dwelling, and on a 'room model' elaboration, which takes into account geometrical and structural characteristics of the dwelling. The validation was performed using data--relevant to 91 dwellings, at present- collected in Latium and Campania within the framework of 'SETIL', the Italian epidemiological study on the aetiology of childhood leukaemia, lymphoma and neuroblastoma. PMID- 15550714 TI - Gamma dose rate calculation and mapping of piemonte (North-West Italy) from gamma spectrometry soil data. AB - In this study the air gamma dose rate map of Piemonte, a region in the North-West of Italy, was produced from gamma spectrometry soil data. Soil samples collected in 110 different sites of Piemonte were analysed with Hyperpure Germanium (HPGe) detectors (30% relative efficiency), which allow the evaluation of the activity concentrations of natural radionuclides and (137)Cs. Then, using the available mathematical models, the gamma absorbed dose rate in air due to radionuclides was calculated. The contribution of the cosmic radiation to the total absorbed dose rate, which depend on the site altitude was also evaluated and added to the soil contribution. Finally, the map of the whole region was obtained by fitting the dose rate values of the different sites with kriging algorithms. PMID- 15550715 TI - Experimental procedures and statistical results for evaluation of exposure to a radiofrequency electromagnetic field with complex analogue modulation. AB - The paper illustrates an experimental set-up used to monitor over long periods the peak and average power associated with a complex analogue modulated signal. The procedure was applied to a standard PAL television transmission. The measurement was carried out from a remote location, and a complete tele-measuring system was designed, configured and tested. Following the application of the measuring system to some real cases, interesting statistical results illustrating long-period exposure values were reported. PMID- 15550716 TI - Exposure measuring techniques for wide band mobile radiocommunications. AB - The paper illustrates the limits and performances of different experimental monitoring techniques, which are applied to digitally modulated radiofrequency electromagnetic fields used for mobile telecommunications. Different experimental set-ups have been developed, verified and applied for the analysis and characterisation of wide band probes and narrow band measuring procedures. PMID- 15550717 TI - Indoor radon concentration forecasting in South Tyrol. AB - In this paper a modern statistical technique of multivariate analysis is applied to an indoor radon concentration data base. Several parameters are more or less significant in determining the radon concentration inside a building. The elaboration of the information available on South Tyrol makes it possible both to identify the statistically significant variables and to build up a statistical model that allows us to forecast the radon concentration in dwellings, when the values of the same variables involved are given. The results confirm the complexity of the phenomenon. PMID- 15550718 TI - Radon mapping in south Tyrol: comparison between two different procedures. AB - In this paper two different procedures for radon mapping have been compared on the same database referring to indoor radon records of South Tyrol. The first procedure is based on descriptive statistics applied to administrative units while the second one implies a more complicated statistical analysis applied to a regular grid: it involves floor level normalization of radon data, filling and smoothing algorithms for the territory cells. PMID- 15550719 TI - Development of numerical phantoms by MRI for RF electromagnetic dosimetry: a female model. AB - Numerical human models for electromagnetic dosimetry are commonly obtained by segmentation of CT or MRI images and complex permittivity values are ascribed to each issue according to literature values. The aim of this study is to provide an alternative semi-automatic method by which non-segmented images, obtained by a MRI tomographer, can be automatically related to the complex permittivity values through two frequency dependent transfer functions. In this way permittivity and conductivity vary with continuity--even in the same tissue--reflecting the intrinsic realistic spatial dispersion of such parameters. A female human model impinged by a plane wave is tested using finite-difference time-domain algorithm and the results of the total body and layer-averaged specific absorption rate are reported. PMID- 15550721 TI - In vitro and in vivo correlation of the inhibitory effect of cyclosporin A on the transporter-mediated hepatic uptake of cerivastatin in rats. AB - Previously, we have shown that the inhibition of the transporter-mediated hepatic uptake of cerivastatin (CER) by cyclosporin A (CsA) could, at least partly, explain a pharmacokinetic interaction between these drugs in humans. In the present study, we have examined the effect of CsA on the in vivo disposition of CER in rats and the in vitro uptake of [14C]CER in isolated rat hepatocytes in an attempt to evaluate the effect of inhibition of transporter-mediated hepatic uptake on the in vivo CER disposition. The steady-state plasma concentration of CER increased 1.4-fold when coadministered with CsA up to a steady-state blood concentration of 4 microM. Studies of [14C]CER uptake into isolated rat hepatocytes showed saturable transport, with the saturable portion accounting for more than 80% of the total uptake. CsA competitively inhibited the uptake of [14C]CER with a Ki of 0.3 microM. The IC50 for the uptake of [14C]CER in the absence and presence of rat plasma was 0.2 and 2.3 microM, respectively. The in vivo hepatic uptake of [14C]CER evaluated by the liver uptake index method was also inhibited by CsA in a dose-dependent manner. On the other hand, CsA did not inhibit the metabolism of [14C]CER in rat microsomes. The in vitro and in vivo correlation analysis revealed that this pharmacokinetic interaction between these drugs in rats could be quantitatively explained by the inhibition of transporter mediated hepatic uptake. Thus, this drug-drug interaction in rats is predominantly caused by the transporter-mediated uptake process. PMID- 15550722 TI - Use of a benzyloxy-substituted lactone cyclooxygenase-2 inhibitor as a selective fluorescent probe for CYP3A activity in primary cultured rat and human hepatocytes. AB - The benzyloxy-substituted lactone cyclooxygenase-2 inhibitor DFB [3-[(3,4 difluorobenzyl)oxy]-5,5-dimethyl-4-[4-(methylsulfonyl)phenyl]furan-2(5H)-one] is metabolized in human and rat liver microsomal incubations and hepatocytes to a fluorescent metabolite, DFH [3-hydroxy-5,5-dimethyl-4-[4 (methylsulfonyl)phenyl]furan-2(5H)-one]. This process is CYP3A-mediated in both species, as demonstrated by incubations with recombinant CYP3A enzymes and experiments with inhibitory antibodies. Measurement of DFH fluorescence can be used as a rapid readout of CYP3A activity following microsomal or cultured hepatocyte incubations. In rat and human hepatocytes treated with prototypical inducers, the formation of DFH was linear for the first 30 min, with no secondary metabolism of DFH, such as phase II glucuronidation, observed at early time points. Using a panel of four prototypical inducers (phenobarbital, dexamethasone, phenytoin, and rifampicin), the correlation between testosterone 6beta-hydroxylation in cultured human hepatocytes and CYP3A enzyme level in cell lysate was confirmed. DFB debenzylation was then shown to correlate well with testosterone 6beta-hydroxylation in hepatocytes treated with these four inducers. Primary cultured rat and human hepatocyte induction assays were optimized for 24- and 96-well plates, respectively. Controls were established to evaluate whether test compounds demonstrate time-dependent CYP3A inhibition to avoid false negative results. Thus, the use of DFB, a fluorogenic CYP3A-selective probe substrate, affords a fast, efficient, and robust assay for the measurement of CYP3A induction in both rat and human cultured primary hepatocytes. PMID- 15550723 TI - Predicting the clearance of CYP2C9 substrates. PMID- 15550725 TI - In vitro study of mycophenolic acid glucuronidation. PMID- 15550727 TI - Superior in-hospital and 30-day outcomes with abciximab versus eptifibatide: a contemporary analysis of 495 consecutive percutaneous coronary interventions. AB - BACKGROUND: Both abciximab (AB) and eptifibatide (EP) are approved for use during percutaneous coronary intervention (PCI) but comparative data between the 2 agents are limited. METHODS: We compared in-hospital and 30-day outcomes of contemporary 495 consecutive PCIs performed by a single operator between July 2001 and November 02 with AB and EP (242 with AB and 253 with EP). Cardiogenic shock patients who underwent a second procedure within 30 days from their first procedure were excluded. Selection of glycoprotein IIb/IIIa was at the operator's discretion. The initial 444 cases were performed with unfractionated heparin and the last 51 with bivalirudin. RESULTS: AB cases comprised a higher risk group with more patients with diabetes, peripheral vascular disease, ST-elevation myocardial infarction and renal failure (p<0.05 for each) and more rotablator use and longer lesions (p<0.01 for each). AB was associated with less in-hospital (4.6 versus 12.3%; OR: 0.34; 95% CI: 0.17-0.7; p=0.004) as well as 30-day (5.5 versus 14%; OR: 0.37; 95% CI: 0.19-0.71; p=0.003) major adverse cardiac events (sum of death, MI, urgent revascularization, all bleeding and stroke). CONCLUSION: Despite its use in higher risk PCI patients, AB resulted in superior short-term outcomes compared with EP. Our data suggest that significant efficacy differences may exist between these 2 agents and that a randomized comparison is warranted. PMID- 15550728 TI - Coronary pressure notch: an early non-hyperemic visual indicator of the physiologic significance of a coronary artery stenosis. AB - The disappearance of a dichrotic notch on the peripheral arterial pulse wave has been associated with significant peripheral vascular disease. A similar observation has not been reported in the distal coronary pressure waveform. The purpose of this study was to investigate the significance of a coronary pressure notch distal to a coronary stenosis and its relationship to fractional flow reserve. Ninety-seven patients with 131 angiographically indeterminate lesions (40-80% diameter narrowing) underwent FFR measurements for physiological significance. Hemodynamic tracings were recorded prior to the administration of adenosine and visually analyzed for the presence or absence of a dicrotic notch in the distal coronary artery pressure tracing. The stenoses were then divided into two groups based on the presence or absence of a notch. Of the 54 lesions without a distal coronary pressure notch, 31 had a FFR greater than or equal to 0.75 and of the 77 lesions with a notch, 75 had a FFR greater than or equal to 0.76. The sensitivity and specificity of a pressure notch was 94% and 74%, respectively, with positive and negative predictive values of 57% and 97%, respectively. The presence of a distal coronary pressure notch was predictive of a FFR greater than or equal to 0.76. The distal dicrotic pressure notch may be used as an additional parameter without requiring hyperemia for FFR measurements of uncertain clinical significance. PMID- 15550729 TI - Primary angioplasty of unprotected left main coronary artery for acute anterolateral myocardial infarction. AB - BACKGROUND: Mortality of acute unprotected left main coronary artery (LMCA) occlusion is very high. The objectives of this analysis were to determine the effect of primary angioplasty and the impact of cardiogenic shock on unprotected LMCA occlusion-induced acute anterolateral myocardial infarction (AAMI). METHODS: Of 1,736 consecutive patients with acute myocardial infarction (AMI), 38 (2.2%) had LMCA occlusion-induced AAMI with Thrombolysis in Myocardial Infarction (TIMI) flow less than or equal to 2. All were given primary angioplasty. RESULTS: Of these 38 patients, 17 (45%) were discharged, and 21 (55%) died in-hospital. Cardiogenic shock was overt in 28 patients; 47.1% of the survival group and 95.2% of the mortality group (p=0.0008). On arrival, the survival-group had higher pH (7.40+/-0.10 vs. 7.30+/-0.14; p=0.013) and base excess (-4.5+/-3.9 vs. -10.4+/ 6.0 mEq/L; p=0.0013). In the survival group reperfusion was successful in 100% of patients, as opposed to 57.1% in the mortality group (p=0.0020), and the incident of stenting was not different between the two groups (64.7% vs. 71.4%, p=0.66). Shock patients had lower successful angioplasty rate (67.9% vs. 100%, p=0.040), higher in-hospital mortality (71.4% vs. 10.0%, p=0.0008), and higher 1-year mortality rates (p=0.0064), than stable patients. All shock patients with failed angioplasty died, but the mortality rate was 57.9% (p=0.021) when angioplasty was successful. CONCLUSIONS: Patients presenting with AAMI, LMCA occlusion, and cardiogenic shock have poor survival regardless of primary angioplasty in conjunction with coronary stents. Nevertheless, primary angioplasty is a feasible and effective procedure, and it may save lives in this clinical setting. PMID- 15550730 TI - PCI for acute myocardial infarction caused by left main disease. PMID- 15550731 TI - Predictors of survival after contemporary percutaneous coronary revascularization for acute myocardial infarction in the real world. AB - Management strategies for ST-elevation myocardial infarction (STEMI) have undergone great evolution over the past decade. The objectives of this study were to evaluate the in-hospital and long-term clinical outcomes, as well as predictors of survival, among patients who received the most contemporary percutaneous coronary revascularization strategies for STEMI in real clinical practice. During the period from October 1, 2000 to April 30, 2002, 316 patients have undergone primary percutaneous coronary intervention (PCI) in a tertiary University hospital, the in-hospital (11.1%), 30-day (13.9%) and long-term (21.8%) mortality rates were higher than that reported in randomized studies. This is likely to be due to the higher prevalence of adverse clinical profiles. Multivariable analysis show that age >65, cardiogenic shock, resuscitated cardiac arrest and intubation independently predicted in-hospital and long-term mortality, while multi-vessel disease predicted major adverse cardiac event (MACE). Among patients with cardiogenic shock, similar mortality was observed in patients with anterior myocardial infarction (MI) or inferior MI with/without right ventricle involvement. PMID- 15550732 TI - Bacteremia following complex percutaneous coronary intervention. AB - BACKGROUND AND OBJECTIVE: Complex percutaneous coronary intervention (PCI) often requires introduction of numerous devices into and out of the arterial circulation and this may result in an increased risk of bacteraemia or even septicaemia. This study was undertaken to detect the frequency of bacteraemia that may be associated with such procedures. METHODS: 147 patients undergoing complex PCI had blood culture tests immediately after and 12 hours after the procedure. RESULTS: Of 147 patients, 26 (17.7%) had detectable bacteraemia immediately after PCI. Coagulase-negative staphylococcus was isolated most commonly. An additional 12% of patients yielded positive blood cultures in the next 12 hours with femoral sheaths still in-situ. There were no associated clinical sequelae. CONCLUSIONS: Uncomplicated bacteraemia is not uncommon as a result of complex PCI procedures. Although there are usually no clinical sequelae, these findings are important for those patients who are considered to be at moderate or high risk of infective endocarditis who require an invasive procedure such as PCI. This paper emphasizes the need for maximum sterility during PCI procedures if infective complications and stent infection are to be avoided. PMID- 15550733 TI - Vasoreactivity of the radial artery after transradial catheterization. AB - The vasomotor response was used to assess the degree of radial artery injury after transradial catheterization. Vasoreactivity was studied by ultrasound before catheterization, 24 hours after, at 1 week and at 1 month in 18 patients. Mean radial artery diameter increased from 2.56+/-0.45 mm before catheterization to 2.86+/-0.48 mm at 24 hours (p=0.001) and returned to baseline values at 1 month (2.60+/-0.27 mm; p=0.95). Hyperemia-induced vasodilation did not change significantly (2.7+/-4.7% at baseline; 3.4+/-3.7% at 24 hours, 3.5+/-3.9% at 1 week and 4.8+/-4.7% at 1 month; p=0.59). Nitroglycerin-induced vasodilation was significantly attenuated at 24 hours (from 14.1+/-7.9% at baseline to 6.5+/-8.4% at 24 hours; p=0.01), but improved after 1 week (9.8+/-8.5%; p=0.1, compared to baseline) and after 1 month (13.0+/-8.9%; p=0.51, compared to baseline). Thus, soon after transradial catheterization vasoreactivity is impaired, but generally recovers as early as 1 month after the procedure. PMID- 15550734 TI - Transradial catheterization: the road less traveled. PMID- 15550735 TI - Six-month clinical and angiographic outcomes of the Tecnic Carbostent coronary system: the phantom IV study. AB - The aim of this multicenter, prospective, non-randomized study was to examine the procedural, in-hospital, and mid-term clinical and angiographic outcome of patients undergoing coronary angioplasty with the Tecnic Carbostent system. Between October 2001 and March 2002, 123 consecutive patients were treated with coronary implantation of the Tecnic Carbostent. Stable angina (54%), unstable angina (37%) and silent ischemia (9%) were clinical indications for revascularisation. The baseline lesion morphology was complex (Type B2 or C) in 59% of the cases, and the mean lesion length was 15+/-8 mm. A total of 179 stents were implanted in 149 lesions. The procedural success rate was 100%. Mean percent diameter of the stenosis decreased after the intervention from 75%+/-11% to 8%+/ 4%. The mean cross-sectional area stent recoil was 8.8%+/-7.3%. No in-hospital or 30-day major adverse cardiac events were observed. During the 6-month follow-up period, there were no deaths or myocardial infarctions, whereas the incidence of target lesion revascularisation was 12.7%. The angiographic restenosis rate was 14.1%: a focal or limited pattern (class I or II) was found in 83% of cases, whereas the remaining 17% had a proliferative morphology (class III or IV). In conclusion, this study indicates that a good clinical and angiographic outcome may be obtained with the Tecnic Carbostent coronary system in consecutive patients with de novo coronary lesions. PMID- 15550736 TI - Primary angioplasty without on-site surgical back-up: the first experience with mobile catheterization facility. AB - BACKGROUND AND OBJECTIVES: The aim of the present study is to assess the safety and efficacy of performing primary angioplasty in a center without on-site surgical back-up, and compare the data with the literature. METHODS: Seventy eight consecutive primary angioplasty procedures, performed in our center from January 2001 to February 2003, were followed prospectively. Clinical and demographic characteristics of the patients, procedural success, early and late outcomes of the patients were taken into account. The safety of angioplasty was assessed by the analysis of in-hospital complications (death, urgent need for repeat revascularization, AMI with or without ST-elevation and stroke). The angioplasty procedures were considered effective when the post-procedural residual stenosis did not exceed 50% with the distal Thrombolysis in Myocardial Infarction (TIMI) grade III flow. RESULTS: The device success rate was 92.3%. Angiographic success rate was 88.8%. In hospital mortality rate was 4.1%. These patients were admitted with cardiogenic shock; 1 died during the procedure and the other 2 died during hospital follow-up. One patient died suddenly and another developed acute MI during the 6-month follow-up period. No patients developed stroke or were referred for urgent surgery. Four patients (5.5%) underwent repeat angioplasty during follow-up. CONCLUSIONS: Primary angioplasty can be safely performed in centers without on-site surgery. The efficacy and safety requirements of angioplasty, performed in a center without on-site surgical back up using a mobile catheterization facility were similar to the data obtained from the literature. PMID- 15550737 TI - Primary PCI without on-site surgery and without on-site cath lab. PMID- 15550739 TI - The Angiomax Peripheral Procedure Registry of Vascular Events Trial (APPROVE): in hospital and 30-day results. AB - BACKGROUND: High-risk patient characteristics and complexity of percutaneous peripheral intervention (PPI) procedures suggest a need for predictable and reliable anticoagulation. We undertook this study to assess the safety and efficacy of bivalirudin as the procedural anticoagulant in patients undergoing PPI of the renal, iliac, or femoral artery. METHODS: This was a prospective, open label, single arm study inpatients undergoing PPI of the renal, iliac, or femoral vessels to assessbivalirudin as the sole procedural anticoagulant (0.75 mg/kg bolus/1.75 mg/kg/hr infusion). The primary endpoint was procedural success defined as residual stenosis < 20%. Secondary endpoints included ischemic events (death, myocardial infarction, unplanned revascularization, and amputation), and bleeding complications, as well as ACT values and times to sheath removal, ambulation, and discharge. RESULTS: 505 patients were enrolled at 26 sites. Procedural success was achieved in 95.0% of patients. Ischemic events were low (1.4%) and similar between vessel types. Protocol-defined major hemorrhage and TIMI major hemorrhage rates were 2.2% and 0.4%, respectively. Mean ACTs were similar among treatment groups (renal 353.8 seconds(s); iliac 335.9s, femoral, 343.5s). CONCLUSION: Bivalirudin provided consistent anticoagulation and similar outcomes in all vessel types treated at the dose tested. Ischemic and bleeding event rates were low, demonstrating the safe use of bivalirudin as a procedural anticoagulant in PPI. PMID- 15550741 TI - Reversible pulmonary hypertension in a patient treated with protacyclin. PMID- 15550740 TI - Acute right coronary artery occlusion in an adult patient after radiofrequency catheter ablation of a posteroseptal accessory pathway. PMID- 15550742 TI - The novel use of a covered stent in the management of a left internal mammary artery to pulmonary vasculature fistula. PMID- 15550743 TI - Distal protection of the left anterior desceding artery with the EX Filter Wire during an acute coronary syndrome. PMID- 15550744 TI - Diffuse saphenous vein graft spasm. PMID- 15550745 TI - Advanced atherosclerotic plaque as a potential cause of no-reflow in elective percutaneous coronary intervention: intravascular ultrasound and histological findings. PMID- 15550746 TI - Successful revascularization of a long chronic total occlusion with blunt microdissection complicated by coronary artery dissection. AB - We report a case of successful percutaneous revascularization of a chronic total occlusion using the LuMend Frontrunner catheter. The case was complicated by a long coronary artery dissection, with inability to access the true lumen. With favorable healing at 7 weeks, the true lumen was accessible which led to procedural success. PMID- 15550747 TI - Transcatheter treatment of Lutembacher syndrome: combined balloon mitral valvuloplasty and percutaneous atrial septal defect closure. PMID- 15550748 TI - Nickel allergy and the amplatzer septal occluder. PMID- 15550749 TI - Unprotected left main "kissing" stent implantation with a percutaneous ventricular assist device. PMID- 15550752 TI - Iron and oxidative stress in renal insufficiency. AB - BACKGROUND/AIMS: Iron (Fe) can cause tissue injury and oxidative stress by catalyzing hydroxyl radical production and lipid peroxidation. Intravenous (i.v.) Fe preparations are routinely administered to treat anemia in patients with chronic renal failure (CRF), a condition marked by oxidative stress and inflammation. In an earlier study, we showed that iron overload augments oxidative stress in the cardiovascular tissues of CRF rats. This study was designed to expand these observations to other major organs. METHODS: Rats were randomized into CRF (5/6 nephrectomized) and sham-operated control (CTL) groups. Each group was subdivided into Fe-loaded (single i.v. injection of iron dextran complex, 0.5 g/kg) and placebo-treated subgroups. After 13 weeks, systolic blood pressure, blood hemoglobin (Hb), plasma Fe concentration, lipid peroxidation products, superoxide generating enzyme, NAD(P)H oxidase, and antioxidant enzymes were determined. RESULTS: Systolic blood pressure was equally elevated and creatinine clearance was equally reduced in both CRF groups. Fe administration raised Hb, serum Fe and transferrin saturation in both CRF and CTL groups. The plasma concentration of lipid peroxidation product, malondialdehyde, was increased by Fe injection in CRF rats but not the control group. Renal tissue abundance of gp91(phox) subunit of NAD(P)H oxidase was elevated in the untreated CRF group and was partially reduced in the iron dextran-treated CRF group. Tissue abundance of the antioxidant enzymes; superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) were decreased in both untreated and iron dextran-treated CRF groups. CONCLUSION: CRF resulted in marked SOD, CAT and GPX deficiencies. A single i.v. administration of iron dextran in rats with CRF induced oxidative stress as measured by increased lipid peroxidation products and decreases in antioxidant enzymes. PMID- 15550753 TI - Efficacy of multidrug therapy combined with mizoribine in children with diffuse IgA nephropathy in comparison with multidrug therapy without mizoribine and with methylprednisolone pulse therapy. AB - AIM: To evaluate the efficacy of prednisolone, warfarin, and dipyridamole therapy combined with mizoribine (PWDM) in the treatment of diffuse immunoglobulin A (IgA) nephropathy in comparison with prednisolone, warfarin, and dipyridamole therapy without mizoribine (PWD) and with methylprednisolone pulse therapy (PWD pulse). METHODS: We collected data on 61 patients diagnosed with diffuse IgA nephropathy, and these patients were retrospectively divided into three groups without randomization. Group A included 21 patients before 1987 who were treated with PWD for 24 months, group B included 20 patients from 1987 to 1989 who were treated with PWD pulse therapy for 24 months, and group C included 20 patients after 1990 who were treated with PWDM for 24 months. Clinical features and pathological findings in each group were analyzed retrospectively. RESULTS: The time from initiation of therapy in group A, group B, and group C was 8.9 +/- 5.2, 8.1 +/- 3.9, and 7.7 +/- 3.8 years, respectively. At the latest follow-up examination, the mean urinary protein excretion (mg/m2/h) was 17 +/- 10 in group A, 22 +/- 20 in group B, and 6 +/- 6 in group C and had decreased significantly in group C as compared with the other groups. The activity index in all three groups was lower at the second biopsy than that at the first biopsy (5.1 +/- 0.8 vs. 6.5 +/- 2.1 in group A, p < 0.05; 5.6 +/- 0.9 vs. 6.6 +/- 1.7 in group B, p < 0.01, and 4.5 +/- 1.0 vs. 6.8 +/- 1.9 in group C, p < 0.01). The chronicity index in groups A and B at second biopsy was higher than at first biopsy (7.3 +/- 1.4 vs. 4.8 +/- 1.0 in group A, p < 0.01, and 8.1 +/- 2.0 vs. 5.3 +/- 0.9 in group B, p < 0.01), but was unchanged in group C. At the latest follow-up examination, 1 patient (4.8%) in group A, 3 patients (15%) in group B, and none (0%) in group C had renal insufficiency. CONCLUSION: These results suggest that PWDM appears to be more effective than PWD or PWD pulse in ameliorating proteinuria and histological severity of patients with IgA nephropathy. PMID- 15550754 TI - Cardiac involvement in facioscapulohumeral muscular dystrophy. AB - Cardiac involvement (CI) in form of myocardial thickening in a patient with genetically confirmed facioscapulohumeral muscular dystrophy (FSHMD) has not been reported. The patient is a 50-year-old male with a tandem repeat size of 17 and 14 kb in the D4Z4 locus on chromosome 4q35. The clinical cardiologic investigation was normal. Blood pressure was 150/90 mm Hg. Funduscopy, 24-hour ambulatory ECG, and 24-hour blood pressure monitoring were normal. ECG showed incomplete right bundle branch block, ST elevation in V2-V4, tall T waves in V3 V5, and hypertrophy signs. Echocardiography revealed left ventricular myocardial thickening of the posterior wall (11.7 mm) and the septum (15.5 mm). In conclusion, CI in genetically confirmed FSHMD may manifest not only as ECG abnormalities but also as left ventricular myocardial thickening. PMID- 15550755 TI - Osteogenic potential of human adipose tissue-derived stromal cells as an alternative stem cell source. AB - Adult bone marrow contains mesenchymal stem cells (bone marrow-derived mesenchymal stem cells; BMSCs) which contribute to the generation of mesenchymal tissue such as bone, cartilage, muscle and adipose. However, using bone marrow as a source of stem cells has the limitation of a low cell number. An alternate source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Human adipose tissue obtained by liposuction was processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). In this study, we compared the osteogenic differentiation of ATSCs with that of BMSCs. Both cell types were cultured in atelocollagen honeycomb-shaped scaffolds with a membrane seal (ACHMS scaffold) for three-dimensional culturing in a specific osteogenic induction medium. Optimal osteogenic differentiation in both cell types, as determined by alkaline phosphatase cytochemistry, secretion of osteocalcin, mineral (calcium phosphate) deposition and scanning electron microscopy, was obtained with the same three-dimensional culture. Furthermore, osteoblastic lining in vivo was examined using ATSC-seeded or BMSC-seeded scaffolds in nude mice. The present results show that ATSCs have a similar ability to differentiate into osteoblasts to that of BMSCs. PMID- 15550756 TI - Cellular reactions of osteoblasts to micron- and submicron-scale porous structures of titanium surfaces. AB - Osteoblast reactions to topographic structures of titanium play a key role in host tissue responses and the final osseointegration. Since it is difficult to fabricate micro- and nano-scale structures on titanium surfaces, little is known about the mechanism whereby the topography of titanium surfaces exerts its effects on cell behavior at the cellular level. In the present study, the titanium surface was structured in micron- and submicron-scale ranges by anodic oxidation in either 0.2 M H3PO4 or 0.03 M calcium glycerophosphate with 0.15 calcium acetate. The average dimensions of pores in the structured surface were about 0.5 and 2 microm in diameter, with roughness averaging at 0.2 and 0.4 microm, respectively. Enhanced attachment of cells (SaOS-2) was shown on micron- and submicron-scale structures. Initial cell reactions to different titanium surfaces, e.g. the development of the actin-containing structures, are determined by the different morphology of the surfaces. It is demonstrated that on either micron- or submicron-structured surfaces, many well-developed filopodia were observed to be primary adhesion structures in cell-substrate interactions, and some of them entered pores using their distinct tips or points along their length for initial attachment. Therefore, porous structures at either micro- or submicrometre scale supply positive guidance cues for anchorage-dependent cells to attach, leading to enhanced cell attachment. In contrast, the cells attached to a smooth titanium surface by focal contacts around their periphery as predominant adhesion structures, since repulsive signals from the environment led to retraction of the filopodia back to the cell bodies. These cells showed well organized stress fibres, which exert tension across the cell body, resulting in flattened cells. PMID- 15550757 TI - Morphological features of tail bud development in truncate mouse mutants. AB - A key malformation in the homozygous truncate mouse mutants is a partial lack of the notochord in the embryo tail. In order to analyze if tail bud development was affected by the truncate (tc) mutation, serial semithin sections of tails of the homozygous mutant embryos were compared to the wild-type controls. In the wild type embryos morphologically uniform mesenchyme of the tail bud was continuous via the medullary cord to the secondary neural tube, and via the tail cord to the notochord and the gut. In truncate embryos the tail cord was not continuous to the notochord, but to the additional lumen of the tail gut resulting in tail gut duplication. Toward the base of the tail two tail guts subsequently fused together or the additional one disappeared. If present in the tip of the tail, the notochord in truncate embryos ended near the ventral border of the secondary neural tube. The rest of the tail notochord was fragmented and the posterior ends of the fragments were frequently adjacent or even continuous to the neural tube. We suggest that the improper regionalization of the tail bud, where notochord is associated with the neural tube rather than with the tail gut, is related to the subsequent segmental lack of the notochord in truncate mutants. PMID- 15550758 TI - Purinergic signalling to rat ovarian smooth muscle: changes in P2X receptor expression during pregnancy. AB - The expression of P2X and P2Y receptor subtypes in the smooth muscle of the rat ovary during the oestrus cycle and pregnancy was examined using immunohistochemistry. RT-PCR studies of P2X receptor mRNA were also carried out. In the non-pregnant rats, P2X2 receptor protein was dominant in the smooth muscle of perifollicular rings and blood vessels. P2X1 protein expression was seen on vascular smooth muscle too, but little, if any, was present on perifollicular smooth muscle. No changes in P2X1 or P2X2 receptor expression were seen during the oestrous cycle. During early and mid-late pregnancy, there was a switch from P2X2 to P2X1 receptor protein expression in the smooth muscle of the perifollicular ring; P2X1 receptors were also more prominently expressed than P2X2 receptors on ovarian vascular smooth muscle in non-pregnant animals, but during late pregnancy the expression of P2X2 receptors was found to equal that of the P2X1 receptors. There was a return to non-pregnant P2 receptor subtype distribution 2 days after birth. Ovarian vascular and perifollicular smooth muscle showed immunoreactivity for P2Y1, but not for P2X3-7, P2Y2 or P2Y4 receptors. P2Y1 receptor expression in ovarian smooth muscle of both blood vessels and follicular rings did not show significant changes during the oestrus cycle or pregnancy. RT-PCR studies indicated that P2X1 and P2X2 receptor mRNA was present in the ovary during pregnant and non-pregnant conditions. P2X4-6 receptor mRNA was also present in all stages studied, however no immunostaining showing receptor protein for these subtypes was seen on the ovarian sections examined. In summary, purinergic signalling to ovarian perifollicular smooth muscle changed from P2X2 to P2X1 receptors during pregnancy, while there was an increase in P2X2 receptor expression on vascular smooth muscle. PMID- 15550759 TI - Changes in the epithelium of the vaginal complex during the estrous cycle of the marsupial Monodelphis domestica. 2. Scanning electron microscopy study. AB - The four stages of the estrous cycle in Monodelphis domestica, namely proestrus, estrus, postestrus and the transitional metestrus, were analyzed with the scanning electron microscope and compared with the results of the previously published transmission electron-microscopic paper [Cells Tissues Organs 2002;172:276-296]. During the estrous cycle the vaginal epithelium undergoes dramatic changes from a nonkeratinized to a highly keratinized epithelium. The predominant feature of proestrus with the beginning of keratinization is the presence of polygonal flat cells with pavement-like appearance, bordered by raised ridges and covered with microvilli. The epithelium is fully keratinized in estrus, and the superficial layers overlap like shingles. Many cells are still densely covered by microvilli, whereas others develop a complex pattern of microridges. In postestrus different epithelial structures are revealed depending on the actual stage of desquamation. In early postestrus surface cells resemble those present during estrus. In late postestrus, when only few keratinized cells are left, the nonkeratinized cells become exposed to the lumen through desquamation. These cells border the lumen during metestrus, a cycle stage during which numerous leukocytes migrate into the vaginal canal. A number of these uppermost cells is probably not yet prepared to function as metestrus cells and are therefore sloughed off as well. During metestrus compact cell masses stick in the vaginal furrows. Epithelial surface cells are highly irregular and bulging with their microvilli covered surfaces in the vaginal lumen. This study represents the first comprehensive description of alterations on the surface ultrastructure of a marsupial vagina during the estrous cycle, demonstrating considerable differences in comparison to many eutherians. PMID- 15550760 TI - Is cerebrospinal fluid spectrophotometry useful in CT scan-negative suspected subarachnoid haemorrage? AB - Missed cerebral aneurysms in CT-negative patients can have serious implications. We set out to determine the usefulness of cerebrospinal fluid (CSF) spectrophotometry and the individual significance of CSF oxyhaemoglobin, bilirubin and methaemoglobin in 463 CT scan-negative patients with suspected subarachnoid haemorrhage (SAH) and normal neurological examination. CSF spectrophotometry resulted in the diagnosis of an intracranial aneurysm in 2% (9/463) of patients with CT-negative suspected SAH. No aneurysms were found in patients in whom spectrophotometry was negative for haem pigments. Less than 1% of patients with oxyhaemoglobin alone had aneurysms diagnosed, whilst 21% of patients with bilirubin had an aneurysm. CSF spectrophotometry is an important investigation in patients with CT-negative suspected SAH, particularly where clinical suspicion is strong. Patients positive for bilirubin are likely to provide a high yield of aneurysmal bleed and should undergo angiography. Patients with oxyhaemoglobin alone in whom SAH is strongly suspected may benefit from angiography. Based on a small number of patients, we recommend that patients with methaemoglobin should also be investigated. Patients with negative spectrophotometry are unlikely to benefit from further investigation. PMID- 15550761 TI - New-onset seizures after liver transplantation: clinical implications and prognosis in survivors. AB - PURPOSE: To identify the probable etiologies and characteristics of new-onset seizures after orthotopic liver transplantation (OLT) and to assess their clinical implications and prognosis. METHODS: We retrospectively analyzed the clinical, electrophysiologic and laboratory data of 17 patients with new-onset seizures after OLT among 367 adult and pediatric patients who underwent OLT between 1999 and 2001. RESULTS: A suspected etiology of seizures was identified in most patients, including 6 (35.2%) with neurotoxicity due to immunosuppressive therapy, 4 (23.5%) with cerebrovascular disease, 3 (17.6%) with severe metabolic derangement by sepsis or rejection, and 1 each (5.8%) with hyperglycemia and brain edema due to fulminant hepatic failure. Causative factors could not be identified in 2 patients (11.8%). Seizures recurred in 15 patients (88.2%), with 9 occurring on the same day as the original seizure. Attacks caused by neurotoxicity tended to have an earlier onset, within 1 week in 4 of 6 patients, than those caused by cerebrovascular disease and sepsis/rejection, but this was not statistically significant. A total of 21 EEGs were performed in 13 patients. Eleven patients had abnormal EEG findings, of whom 4 (30.7%) showed epileptiform discharges, but the outcome of patients with epileptiform activity did not differ statistically from that of patients without such discharges (p > 0.6). The incidence of poor outcome (death or persistent vegetative state) in the group with seizures was almost 10 times higher than in the group without seizures (52.9 vs. 5.7%, p < 0.001). The prognosis of patients with seizures due to cerebrovascular disease and severe metabolic derangement by sepsis/rejection was poorer than that of patients with seizures caused by the neurotoxicity of immunosuppressive drugs (p < 0.02), suggesting that the underlying cause of seizures is important in determining prognosis. Of 8 patients who survived, 1 was lost to follow-up. The long-term outcome of seizures in surviving patients was excellent, with all survivors available for follow-up being seizure-free for a mean follow-up of 42.5 months (range, 16-58 months). CONCLUSION: New-onset seizures after OLT may herald fatal outcome, especially in patients with cerebrovascular disease or sepsis. The prognosis of seizures in survivors is excellent, and long-term antiepileptic drugs are not required in most cases. PMID- 15550762 TI - Steroid therapy altered serum levels of CCL2 and CCL5 chemokines in multiple sclerosis patients during relapse. AB - Chemokines are involved in the pathogenesis of multiple sclerosis. The aim of this study was to determine the impact of steroid therapy on the levels of CCL2 and CCL5 chemokines. The study encompassed 30 patients with clinically definite relapsing-remitting multiple sclerosis who were treated with methylprednisolone due to the relapse of the disease. The control groups consisted of 20 patients during the stable stage of relapsing-remitting multiple sclerosis and of 15 patients with noninflammatory diseases of the nervous system. Both chemokines were markedly expresssed in serum by enzyme-linked immunosorbent assay. During relapse, the levels of both chemokines differed significantly from the levels measured in both control groups. After the methylprednisolone treatment, the chemokine levels changed significantly: the levels of CCL2 increased, whilst the levels of CCL5 decreased. These alterations did not correlate with the clinical state of the patients or with the therapeutic effect of the treatment and indicated that the inflammatory reaction accompanying the relapse was receding. PMID- 15550763 TI - The pattern of inflammation and a potential new clinical meaning and usefulness of C-reactive protein in end-stage renal failure patients. AB - Inflammatory indexes are frequently elevated in end-stage renal failure (ESRF) patients. It seems that the pattern of inflammation is particular in this population. In the presence of a higher than normal microinflammatory background (CRP, C-reactive protein, values between 0.1 and 10-15 mg/l) that varies with time, waves of 'true' inflammation (CRP > 10-15 mg/l), mainly due to infections, are added periodically. To accurately assess the average microinflammation in these patients, multiple CRP measurements are required. As recent experimental studies showed that inflammation and particularly elevated CRP levels may be risk factors and not just a risk index for atherosclerosis, in this case, the characteristic inflammation pattern might be of importance in the evolution of this disease in ESRF patients. The causes of the inflammatory state in ESRF patients are multiple: renal insufficiency per se and its complications, coexisting diseases, established atherosclerosis, the consequences of renal replacement treatment, and frequent infections are potentially the main ones. The fluctuating inflammatory pattern is probably due to destabilization or changes in time of the above-mentioned parameters. Thus, the clinical meaning of the average microinflammation in these patients, as assessed by CRP measurements, seems to be that of an index indicative of the grade of their health aggravation by the multiple factors implicated in the inflammation formation. CRP is a sensitive, but not specific, risk index of the overall morbidity and mortality in these patients. The manipulation of the inflammation in ESRF patients should include follow-up and treatment of all the factors that contribute to this state and probably medications such as the statins. If inflammation and CRP in particular definitely prove to be risk factors for atherosclerosis, intensification of this treatment will be necessary. PMID- 15550764 TI - Neurogenic inflammation and pancreatitis. AB - Stimulation of primary sensory neurons produces local vasodilation, plasma extravasation, and pain and is due largely to the release of the tachykinins substance P and calcitonin-gene-related peptide. Pathological activation of sensory neurons and the inflammatory sequelae are known as neurogenic inflammation and appear to be important in many organ systems, including the pancreas. Factors that stimulate primary sensory neurons include hydrogen ions, heat, leukotrienes, arachidonic acid metabolites, bradykinin, and proteases such as trypsin, all of which may participate in the generation of acute pancreatitis. The current review examines the cellular and molecular mechanisms involved in sensory nerve activation within the pancreas and the potential contribution of neurogenic inflammation to the pathogenesis of pancreatitis. PMID- 15550765 TI - The role of extended resection in pancreatic adenocarcinoma: is there good evidence-based justification? AB - Thus far, there are no studies concerning the radicality of pancreaticoduodenectomy which, in well-performed, randomized-controlled trials employing high standards of evidence-based medicine, show a benefit over extended lymphadenectomy. The results of the only two prospective randomized studies are not comparable and both are underpowered (level of evidence Ib). Therefore, it is still unclear whether extended lymphadenectomy for pancreatic carcinoma improves outcome. Only one study suggests a positive tendency toward increased survival rates in node-positive patients. Extended approaches including additional venous resection can be performed without a rise in the morbidity and mortality rates of patients with pancreatic carcinoma. In the future appropriately powered randomized trials of standard vs. extended resections may show the benefit of extended surgical resections. In addition, well powered trials of postoperative adjuvant therapies or preoperative neoadjuvant strategies together with surgical resections may identify more effective combinations showing a survival benefit in patients with pancreatic carcinoma. PMID- 15550766 TI - Cell death pathways in pancreatitis and pancreatic cancer. AB - The understanding of the regulation of apoptosis and necrosis, the two principal cell death pathways, is becoming exceedingly important in investigations of the pathogenesis and treatment of pancreatitis and pancreatic cancer. For example, in acute pancreatitis significant amounts of pancreatic necrosis are associated with increased morbidity and mortality. Thus, determining the key steps regulating necrosis should provide insights into potential therapeutic strategies for improving outcome in these patients. On the other hand, in pancreatic cancer various survival mechanisms act to prevent cell death, resulting in promotion of tumor growth and metastasis. Resistance of pancreatic cancer to apoptosis is the key factor preventing responses to therapies. Investigations of the regulation of cell death mechanisms specific to pancreatic cancer should lead to improvements in our current therapies for this disease. The present review is designed to provide information about cell death pathways in pancreatitis and pancreatic cancer with reference to areas that need further investigation, as well as to provide measurement techniques adapted to pancreatic tissue and cells. PMID- 15550767 TI - Sequence analysis of an Asian isolate of minute virus of canines (canine parvovirus type 1). AB - Minute virus of canines (MVC), also known as canine parvovirus (CPV) type 1, is an autonomous parvovirus that infects domestic dogs worldwide and responsible for clinical problems of neonates and pregnant bitches. It was preliminary described that MVC is antigenically as well as genetically different from CPV type 2 that emerged later. However, much of MVC is still obscure since only a limited number of MVC isolates have been available for the study. MVC infections of Japanese and Korean dogs were epidemiologically studied in the previous study, and several MVC isolates could be cultivated in vitro. In the present study an almost full-length nucleotide sequence of a Korean MVC strain HM-6 genome was obtained and comparatively analyzed with those of the American MVC strain GA3 characterized recently. Genome structure of the HM-6 strain was similar to the GA3 strain showing 96.4% identity at the nucleotide sequence. Each of the deduced amino acid sequences of NS1, NP-1, and VP1/2 showed homology of 96.5%, 92.5%, and 97.5% between the HM-6 and GA3 strains. When compared with other parvovirus species, the HM-6 strain was most closely related to bovine parvovirus (BPV) as described for the GA3 strain. These results suggest that MVC together with BPV can possibly be classified into a new clade in the Parvovirinae subfamily. PMID- 15550768 TI - The gene product specified by a double stranded RNA in the flax rust (Melampsora lini) is expressed during rust growth. AB - Strain SP6 of the flax rust (Melampsora lini) contains 11 double-stranded RNAs (dsRNAs) of unknown function. A large open-reading frame (B3ORF1 in dsRNA B3 encodes a polypeptide of 614 amino acids, and using an antiserum raised against the B3ORF1-glutathione S-transferase fusion protein prepared from a bacterial expression system, we have detected the presence of a 67 kDa polypeptide in rust urediospores. This polypeptide, identical in size to that of the predicted translation product of B3ORF1 , was not detected in spores from either a fungal strain lacking the B3 dsRNA or an isogenic strain containing no dsRNA. These data indicate that B3ORF1 present in the flax rust B3 dsRNA is expressed in vivo which warrants farther investigation in search for its function during rust development. PMID- 15550769 TI - Molecular characterization of two distinct begomoviruses from Papaya in China. AB - Six papaya samples showing downward leaf curling were collected in Guangdong and Guangxi provinces, China. The result of TAS-ELISA showed they were all infected by geminiviruses. Comparison of partial DNA-A sequences reveals that these virus isolates can be classified into two groups. Group I includes isolates G2, G4, G5, G28 and G29 from Guangxi province, while isolate GD2 from Guangdong province belongs to Group II. The complete DNA-A sequence of G2 and GD2 were characterized. Sequence comparisons showed that the DNA-A of G2 and GD2 were most closely related to that of Ageratum yellow vein China virus- [Hn2] and Ageratum yellow vein virus , respectively, with 83.4 and 75.2% nucleotide sequence identity, while DNA-A sequence between G2 and GD2 had only 73.4% sequence identity. The molecular data suggests that G2 and GD2 are two distinct begomoviruses, for which the name Papaya leaf curl China virus (PaLCuCNV) for G2 and Papaya leaf curl Guangdong virus (PaLCuGDV) for GD2 are proposed. Comparison of individual encoded proteins showed the coat protein of G2 and GD2 shared highest amino acid sequence identity (97.7 and 94.2%, respectively) with that of Pepper leaf curl virus -[Malaysia] (PepLCV-[MY]), suggesting the CP of these viruses may have identical ancestor. PMID- 15550770 TI - A short insert in the leader sequence of RNA 3L, a long variant of Alfalfa mosaic virus RNA3, introduces two unidentified reading frames. AB - N20-RNA 3L, a large form of RNA 3 associated with Alfalfa mosaic virus (AMV) strain N20 comprises 2281 nt and has approximately 97% overall sequence similarity to the longest previously described RNA 3 of AMV strain YSMV (YSMV-RNA 3; 2188 nt). Compared with YSMV-RNA 3, N20-RNA 3L contains an additional 97 nt in the 5' leader upstream of the open reading frames for movement protein (MP) and coat protein (CP). Two overlapping unidentified reading frames (URF1 and URF2) result from this modification, each of which code for putative translation products of 21 amino acids. The URF1 putative peptide has a hydrophilic N terminus and a hydrophobic C-terminus, indicating a possible association with both host cell membrane and cytosol whereas the putative URF2 product is predominantly hydrophobic. A further structural modification found in N20-RNA 3L is a new tandem repeat of 243 nts which overlaps with the MP open reading frame. PMID- 15550771 TI - Translation of varicella-zoster virus genes during human ganglionic latency. AB - Immunohistochemical analysis of fixed tissue sections of human trigeminal ganglia (TG) and dorsal root ganglia (DRG) revealed the neuronal expression of proteins encoded by Varicella-zoster virus (VZV) genes 21, 29, 62 and 63. These proteins were detected mainly in the neuronal cytoplasm, are likely to be present in low abundance during VZV latency, and mirror the profile of VZV gene transcription. PMID- 15550772 TI - Sequence analysis of the glycoproteins of Tomato chlorotic spot virus and Groundnut ringspot virus and comparison with other tospoviruses. AB - The tospoviruses Tomato chlorotic spot virus (TCSV) and Groundnut ringspot virus (GRSV) cause high economic losses in several vegetable crops in Brazil. The glycoprotein precursor coding sequence was still not available for these two viruses. In this study, the 3' 4 kb M RNA of TCSV and GRSV genome was cloned and sequenced. The sequences were compiled with the available 5' region sequence (NSM gene and 5' UTR) of the same isolates. The M RNA of TCSV was deduced as formed by 4,882 nucleotides, while of GRSV by 4,855 nucleotides. Both M RNA comprised two ORFs in an ambisense arrangement. The vcRNA ORF coded for viral glycoprotein (G1/G2) precursor of TCSV (128.46 kDa) and for glycoprotein precursor of GRSV (128.16 kDa). Comparison of the TCSV and GRSV glycoprotein precursor proteins with those of other tospoviruses showed the highest identity with Tomato spotted wilt virus (81 and 79%, respectively). The amino acid sequence comparison of glycoprotein precursor between TCSV and GRSV revealed a high identity of 92%. However, the nucleotide sequence of the M RNA intergenie region showed only 78%. Phylogenetic analysis was done based on glycoprotein precursor and on M RNA intergenic region of tospoviruses and parameters on tospovirus taxonomic classification were discussed. PMID- 15550773 TI - Genetic conservation of hemagglutinin gene of H9 influenza virus in chicken population in Mainland China. AB - The hemagglutinin (HA) genes of 12 H9N2 influenza virus strains isolated from chickens in Mainland China during the period 1995-2002 were genetically analyzed. All the isolates possessed the same amino acid motif -R-S-S-R/G-L- at the cleavage site of HA. Except for the conserved amino acids, as is the case in the other avian influenza viruses, located in the receptor binding site, all of the 12 isolates possessed N at amino acid position 183; A, T, or V at position 190; K at position 137, whereas the representative strains of the other lineage (except Dk/HK/Y280/97-like lineage) virus of H9N2 viruses had H, E, and R at these positions respectively. These could be considered as the partial molecular markers of the H9 viruses isolated from chickens in Mainland China. Phylogenetic analyses showed HA genes of these isolates belonged to that of A/duck/Hong Kong/Y280/97-like virus lineage. No A/quail/Hong Kong/Gl/97-like virus was found in chicken, population since the outbreak of H9N2 influenza in Mainland China in 1992. The available evidence indicates that HA genes of H9 influenza virus circulating in Mainland China during the past years were well conserved. PMID- 15550774 TI - The TATGARAT box of the HSV-1 ICP27 gene is essential for immediate early expression but not critical for efficient replication in vitro or in vivo. AB - We constructed a recombinant virus containing a promoter mutation altering the immediate-early expression of the HSV-1 ICP27 transcript, ICP27DeltaSma, which contains a deletion of the "TATGARAT" and surrounding sequences, but retains the rest of the ICP27 promoter. This mutant does not exhibit immediate-early expression of ICP27 using criteria of expression in the absence of de novo protein synthesis and earliest expression in the kinetic cascade. While transcript abundance at 1h after infection at 0.1 PFU/cell in mouse embryo fibroblasts was significantly altered compared to infections with wt -rescues, by 4 h after infection these differences were diminished or absent. Consistent with this observation, levels of some critical proteins were reduced in the mutant as compared to rescue infections at the earliest times tested, but were equivalent by 8-12 h pi. Further, both single and multi-step virus replication was equivalent with both mutants and rescues. Thus, altering the immediate early kinetics of ICP27 leads to a sub-optimal quantitative lag-phase in gene expression but without consequence to replication fitness in vitro . Infections in vivo also revealed the ability of mutant and rescue virus to invade the CNS of mice following footpad injections was equivalent. The nature of the role of immediate-early ICP27 expression is discussed in light of these observations. PMID- 15550775 TI - Identification, expression and phylogenetic analysis of the Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV) Helicase gene. AB - The helicase gene from Anticarsia gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV) was identified and localized in the 58.85-65.90 m.u. of the viral genomic map. This gene encodes a putative polypeptide of 1221 amino acids containing motifs homologous to those found in the helicase superfamily. Expression of the AgMNPV helicase was observed as early as 4h post-infection (p.i.) up until 10 h p.i. A unique early transcription initiation site was observed upstream a putative TATA box. Phylogenetic analysis of the helicase genes of 23 baculoviruses indicated that the AgMNPV helicase is closely related to the helicase genes from Epiphyas postvitanna multicapsid nucleopolyhedrovirus and Choristoneura fumiferana defective nucleopolyhedrovirus. PMID- 15550776 TI - Evidence for presence of types A and B of beet necrotic yellow vein virus (BNYVV) in Iran. AB - Rhizomania a viral disease, caused by beet necrotic yellow vein benyvirus (BNYVV), is now widely spread, throughout the sugar beet growing areas of Iran. Genomes of BNYVV are composed of five RNA molecules with specific functions. In this study sequence analyses were conducted on the major coat protein gene (CP21), and parts of RNA3 and RNA4 of an Iranian strain of BNYVV from the Fars province. Sequence alignments of Iran Fars CP21 with other isolates showed closed similarities at nucleotide and amino acid levels with BNYVV pathotype A isolates; S from Japan, and YU2 from Yugoslavia. These results suggest that Iran-Fars isolate probably originated from Asia or neighboring European countries rather than from Germany or France. PMID- 15550777 TI - Cellular source of the poxviral N1R/p28 gene family. AB - Full-length poxvirus N1R/p28 orthologous proteins feature a prominent C-terminal RING zinc-finger motif. The RING moiety is conspicuously mutated in a number of vaccinia virus strains relative to variola virus. This, together with empirical data, suggests that N1R/p28 proteins promote virulence by suppressing apoptosis. Poxvirus N1R/p28 orthologues are strikingly similar to the RING motif of the cellular Makorin family of zinc-finger proteins, suggesting a homologous relationship connecting the viral and cellular genes. Recently identified avipox N1R/p28 orthologues further encode additional Makorin-like zinc-finger motifs, consistent with this suggestion. Phylogenetic analysis supports a model of poxviral capture of a MKRN cDNA and fusion with an existing viral gene. Establishing an evolutionary link between the viral and cellular genes will facilitate the elucidation of their respective cellular functions, and of how they interact in modulating virulence. PMID- 15550778 TI - Putative replication intermediates in endornavirus, a novel genus of plant dsRNA viruses. AB - Oryza sativa endornavirus (OSV) belongs to a new genus (Endornavirus) and family (Endoviridae) with members containing large double-stranded RNA (dsRNA) replicons with plasmid-like properties. Analysis of products obtained from in vitro reaction of the OSV RNA-dependent RNA polymerase revealed a rapid increase of a population of the non-coding strand RNA molecules with a head-to-tail composition. Northern hybridization of total RNA from OSV-carrier cells with riboprobes specific for the coding strand RNA, revealed two types of RNA molecules (i) with a site specific nick and (ii) full-length unnicked molecules. Quantitative analyses of these RNAs showed about 50-fold higher amounts of full length unnicked molecules in cultured cells in which the OSV copy number increases compared with those found in the seedling cells. Both the head-to-tail linked non-coding strand and the full-length coding strand molecules were also found in wild rice and broad beans infected with other endornaviruses indicating that the presence of these unique types of RNA molecules should be considered as a characteristic feature of Endoviridae . PMID- 15550779 TI - Effects of high- and low-dose amiodarone on mortality, left ventricular remodeling, and hemodynamics in rats with experimental myocardial infarction. AB - We examined the effect of high- (AHD) and low-dose (ALD) amiodarone on survival, hemodynamics, and left ventricular remodeling in rats with experimental myocardial infarction (MI). Thirty minutes after coronary artery ligation or sham operation, amiodarone (100 or 20 mg/kg/d) or placebo was given by gavages daily for 8 weeks. Eight weeks later, hemodynamic measurements were performed and left ventricular (LV) volume was determined after KCl-induced cardiac arrest. Early after MI, mortality was lower after both doses of amiodarone. However, excess mortality beginning 15 days after MI outweighed reduced early mortality in rats treated with AHD. Body weight and heart rate were reduced significantly and maximal stroke volume index improved by AHD. In rats with MI, AHD significantly shifted LV pressure-volume curves to the right and increased LV operating volume (2.84 +/- 0.10 versus 2.20 +/- 0.07 mL/kg, P < 0.05). In conclusion, high-dose amiodarone aggravated LV remodeling in rats with large experimental chronic MI probably by lowering heart rate. An early beneficial effect on mortality was probably also lost later by this mechanism. Low-dose amiodarone improved survival without effect on LV remodeling. PMID- 15550780 TI - Mechanisms underlying the sympathomimetic cardiovascular responses elicited by gamma-hydroxybutyrate. AB - Gamma-hydroxybutyrate (GHB) is generally thought to be a central nervous system depressant; however, GHB also has sympathomimetic cardiovascular actions. Radio telemetry was used to record the cardiovascular responses elicited by GHB (180 1000 mg/kg IV) in conscious rats. GHB elicited increases in mean arterial pressure (MAP) (24 +/- 3 to 60 +/- 5 mm Hg) lasting from 28 +/- 8 to 227 +/- 37 minutes. GHB (560 and 1000 mg/kg IV) also elicited a prolonged tachycardic response (85 +/- 23 and 95 +/- 22 bpm). The hypertension and tachycardia elicited by GHB (560 mg/kg) were reversed by the intravenous and intracerebroventricular administration of the GABAb receptor antagonist CGP 35348. CGP 35348 also reversed GHB-mediated increases in renal sympathetic nerve activity (RSNA). Administration of the purported GHB receptor antagonist NCS-382 reversed the increase in heart rate but not the pressor response elicited by GHB in telemetered rats. These data indicate that the intravenous administration of GHB markedly increases MAP, heart rate, and RSNA in conscious rats via activation of central GABAb receptors. In addition, GHB receptors appear to selectively mediate the increase in heart rate elicited by large doses of GHB. PMID- 15550781 TI - Mexiletine protects myocardium during acute ischemia by opening sarcolemmal K-ATP channel: studies in closed-chest acute ischemia model in rabbits. AB - OBJECTIVES: Although we have previously shown that mexiletine might protect myocardium during acute ischemia, the precise mechanism was unclear. In the present study, the mechanism of this effect was examined by using selective K-ATP channel blockers in closed-chest acute ischemia model in rabbits. METHODS: In 40 rabbits, the large left ventricular branch (LLVB) of the left coronary artery was occluded for 30 minutes by inserting a catheter bead (varphi0.5-0.7 x 1.5 mm) through the left carotid artery and was then reperfused. The rabbits were divided into the following 5 groups: (1) control group (n = 8); (2) mexiletine (Mex) group (n = 8, continuous infusion of Mex 24 mg/kg/h); (3) Mex + 5 hydroxydecanoate (5HD) group (n = 8, preadministration of 5HD, 5 mg/kg, followed by Mex infusion); (4) Mex + HMR1098 (selective sarcolemmal K-ATP channel blocker) group (n = 8, preadministration of HMR1098, 3 mg/kg, followed by Mex infusion); and (5) pilsicainide (Pil) group (n = 8, continuous infusion of Pil 18 mg/kg/h). The incidence of ventricular arrhythmia, hemodynamics, left ventricular ejection fraction (LVEF), and infarction size were evaluated and compared among the 5 groups. RESULTS: The incidence of ventricular arrhythmia was lower in groups treated with Mex than the control. The hemodynamics did not show significant differences among the 5 groups. Although the LVEF at 30 minutes after reperfusion was lower in the Mex group (41 +/- 3%, P < 0.001) than the control group (48 +/- 3%), the LVEF at 360 minutes after reperfusion had recovered and became higher in the Mex group (62 +/- 3%, P < 0.001) than the control group (55 +/- 3%). The infarction size was smaller in the Mex group (30 +/- 5%, P = 0.028) than the control group (51 +/- 8%). These effects of Mex were negated by HMR1098 but not by 5HD and were larger than the effects of Pil. CONCLUSIONS: Mex showed improvement in the LVEF in the later phase after reperfusion as well as a reduction in ventricular arrhythmia. The cardioprotective effect of Mex was considered to appear through its action on the sarcolemmal K-ATP channel. PMID- 15550782 TI - Caspase inhibition attenuates contractile dysfunction following cardioplegic arrest and rewarming in the setting of left ventricular failure. AB - Hyperkalemic cardioplegic arrest (HCA) and rewarming evokes postoperative myocyte contractile dysfunction, a phenomenon of particular importance in settings of preexisting left ventricular (LV) failure. Caspases are intracellular proteolytic enzymes recently demonstrated to degrade myocardial contractile proteins. This study tested the hypothesis that myocyte contractile dysfunction induced by HCA could be ameliorated with caspase inhibition in the setting of compromised myocardial function. LV myocytes were isolated from control pigs (n = 9, 30 kg) or pigs with LV failure induced by rapid pacing (n = 6, 240 bpm for 21 days) and were randomized to the following: (1) normothermia (2003 myocytes), incubation in cell culture medium for 2 hours at 37 degrees C; (2) HCA only (506 myocytes), incubation for 2 hours in hypothermic HCA solution (4 degrees C, 24 mEq K); or (3) HCA + z-VAD, incubation in hypothermic HCA solution supplemented with 10 microM of the caspase inhibitor z-VAD (z-Val-Ala-Asp-fluoromethyl-ketone, 415 myocytes). Inotropic responsiveness was examined using beta-adrenergic stimulation (25 nM isoproterenol). Ambient normothermic myocyte shortening velocity (microm/s) was reduced with LV failure compared with control values (54 +/- 2 versus 75 +/- 2, respectively, P < 0.05). Following HCA, shortening velocity decreased in the LV failure and control groups (27 +/- 5 and 45 +/- 3, P < 0.05). Institution of z-VAD increased myocyte shortening velocity following HCA in both the LV failure and control groups (49 +/- 5 and 65 +/- 5, P < 0.05). Moreover, HCA supplementation with z-VAD increased beta-adrenergic responsiveness in both groups compared with HCA-only values. This study provides proof of concept that caspase activity contributes to myocyte contractile dysfunction following simulated HCA. Pharmacologic caspase inhibition may hold particular relevance in the execution of cardiac surgical procedures requiring HCA in the context of preexisting LV failure. PMID- 15550784 TI - Dose-dependent vasodilatory effects of acetylcholine and local warming on skin microcirculation. AB - The assessment of the skin microvasculature response to iontophoretically applied acetylcholine (ACh) and local warming makes it possible to explore noninvasively in humans the functional integrity of endothelium. The present study aimed to examine whether these two stimuli of nitric oxide (NO) release have a dose dependent vasorelaxant activity. For this purpose we assessed in healthy subjects using a laser-Doppler imaging system the increase in forearm blood flow following transdermal application of increasing amounts of ACh [with an iontophoretic current of either 0.28 mC/cm2 (n = 18), 0.56 mC/cm2 (n = 14), 1.4 mC/cm2 (n = 26), 7 mC/cm2 (n = 14), 28 mC/cm2 (n = 14), or 48 mC/cm2 (n = 6)] or graded warming of the skin [to either 37 degrees C (n = 8), 39 degrees C (n = 4) or 41 degrees C (n = 12)]. The maximal vasodilation was significantly smaller with the lowest dose than with the higher doses of ACh, and a plateau was reached with the 1.4 mC/cm2 dose. The skin blood flow responses to ACh were not dependent on the pulsed or continuous pattern of iontophoretic administration. The hyperemia induced by the local heating to 41 degrees C was significantly greater than that observed with the other temperatures. When measured in the same subjects, the magnitude of the maximal ACh-mediated skin blood flow increase was significantly smaller than the vasodilation associated with the warming to 41 degrees C. In summary, transdermally applied ACh and local heating of the skin induce a dose dependent vasorelaxation. These techniques represent a unique means to investigate noninvasively the functional vasodilatory capacity of the skin microvasculature. PMID- 15550783 TI - The cardiovascular and renal effects of the potent and highly selective mu opioid agonist [Dmt1]DALDA. AB - The cardiovascular and renal effects of a mu opioid agonist, [Dmt]DALDA, were studied in conscious Sprague-Dawley rats. During the first hour postinjection, [Dmt]DALDA (0.025-250 microg/rat, IV) evoked a dose-dependent diuresis. The dose of 2.5 microg increased urine volume from 1.0 +/- 0.2 to 3.4 +/- 0.3 mL/h (P < 0.001, n = 30), urinary excretion of sodium, potassium, and cGMP, and induced a mild antihypertensive effect. This dose increased cumulative 4-hour urine volume but significantly inhibited sodium and potassium excretions. The renal and cardiovascular effects were abolished by naloxone (4 mg/kg), but not by naloxonazine (35 mg/kg SC), a selective mu-1 receptor antagonist. Pretreatment with 8 mg/kg naloxone methiodide, an opioid antagonist with limited access to the brain, partially inhibited the renal effects of [Dmt]DALDA. Inhibition of nitric oxide synthases with L-NAME (1 mg/kg) had no effect on the renal and cardiovascular actions of [Dmt]DALDA. Plasma ANP and AVP, measured at 20 and 120 minutes after injection, were not altered by 2.5 and 25 microg [Dmt]DALDA. Therefore, [Dmt]DALDA evokes renal and cardiovascular effects that may primarily be mediated by central naloxonazine-insensitive mu opioid receptors (non-mu-1). These findings indicate that the central mu opioid system is involved in the regulatory mechanism of renal handling of sodium and water. PMID- 15550785 TI - Macrophage inhibitor, semapimod, reduces tumor necrosis factor-alpha in myocardium in a rat model of ischemic heart failure. AB - Pharmaceutical agents aimed at reducing tumor necrosis factor-alpha (TNF-alpha) levels appeared to be attractive possibilities in the medical management of heart failure, as heart failure was shown to be associated with high TNF-alpha levels. However, therapies specifically targeting TNF-alpha failed to show any survival benefit. We examined whether a broad inhibition of inflammatory cytokine production secondary to macrophage inhibition would be more effective at improving cardiac function in the setting of heart failure. To this end, we studied Semapimod (formerly known as CNI-1493), a synthetic guanylhydrazone that inhibits macrophage activation and the production of several inflammatory cytokines. Left anterior descending coronary ligation surgery was performed on each animal to induce a myocardial infarction. After confirming heart failure by echocardiography, the animals were randomly assigned to one of four groups: (1) rats with myocardial infarct receiving high-dose Semapimod, 10 mg/kg/d (MI-H, N = 13); (2) rats with myocardial infarct receiving low-dose Semapimod, 3 mg/kg/d (MI L, N = 9); (3) rats with myocardial infarct receiving vehicle treatment, 2.5% mannitol in water (MI-0, N = 9); and (4) control rats with sham operation and vehicle treatment (Sham, N = 10). Both Semapimod and vehicle treatments were administered by daily tail vein injections over a course of five days. Echoes were repeated at 2, 5, and 9 weeks following treatment. At 9 weeks, hemodynamic data were collected and the animals were euthanized. Trichrome staining was done to assess infarct, and immunohistochemistry was performed to assess TNF-alpha levels. Prior to drug administration, serum was taken from 5 random rats. No detectable level of TNF-alpha was seen (lower limit of detection for the assay used = 12.5 pg/mL). Also prior to any treatment, echocardiography confirmed significant cardiac impairment of rats undergoing LAD ligation as compared with sham. Over the course of the 9 weeks, there were 4 deaths, all in the MI-H group. There was no difference between Semapimod-treated animals and vehicle-treated MI animals in any echocardiographic or hemodynamic parameter. TNF-alpha staining in the noninfarcted region was evident only in the MI groups, not the sham group. When blindly compared on a semiquantitative scale (ie, 0 = no visible staining to 3 = marked staining), a significant difference in staining was observed between MI-0 versus MI-H (1.19 +/- 0.32 versus 0.33 +/- 0.14; P = 0.03) and between MI-0 and MI-L (1.19 +/- 0.32 versus 0.39 +/- 0.22; P = 0.05). In this setting, despite the fact that Semapimod treatment decreased tissue TNF-alpha levels, it did not improve cardiac function, and at high doses it was associated with higher mortality. These results in a rodent model confirm the results of clinical trials with etanercept and infliximab (ie, that decreasing TNF levels in plasma or tissues does not improve cardiac function and may actually increase mortality). PMID- 15550786 TI - Different effects of L-type and L+N-type calcium channel blockers on hamster cheek pouch venules. AB - Dihydropyridine calcium channel blockers are not uniform in terms of their action on calcium channel. L-type calcium channel blockers dilate the resistance arterioles. Cilnidipine is a dihydropyridine calcium channel blocker that also acts on N-type calcium channels, and may dilate venules through its effect on the sympathetic receptor. The influence of an L-type calcium channel blocker (nifedipine) or this L+N type blocker at 10(-7) mol to 10(-4) mol on venular diameter was examined by superfusion of male Syrian hamster cheek pouches. Nifedipine dose dependently dilated the arterioles alone, whereas cilnidipine dilated both arterioles and venules. Application of 10(-7) mol omega conotoxin, an inhibitor of N-type channels, after nifedipine led to significant dilation of venules, while it had no influence when administered after cilnidipine. These findings indicate that the effects of calcium channel blockers on the venules differ according to the action on N-type calcium channels, and that cilnidipine (an L+N type calcium channel blocker) dilates venules through its additional action on N-type channels. PMID- 15550787 TI - The differential role of exogenous and endogenous prostacyclin in the control of renin release from dog renal cortical slices. AB - Using a continuous superfusion system of dog renal cortical slices, we studied the role of prostacyclin in the control of renin release. Superfusate renin activity and prostacyclin as 6-keto-prostaglandin F1alpha, a stable metabolite of prostacyclin, concentrations were measured by radioimmunoassay. Exogenous prostacyclin (0.1, 1, 10 microM) produced a concentration dependent and significant increase in renin release. The calcium ionophore A23187 (10 microM) produced a significant increase in 6-keto-prostaglandin F1alpha release and a significant decrease in renin release. A23187 (10 microM) hardly produced changes of 6-keto-prostaglandin F1alpha release and renin release in the absence of Ca2+. Pretreatment with indomethacin (10 microM) completely abolished the stimulatory effect of A23187 (10 microM) on 6-keto-prostaglandin F1alpha release. On the other hand, the inhibitory effect of A23187 on renin release in the pretreatment with indomethacin was almost equal to that in the "untreatment" with indomethacin. Moreover, we found that there was no association of 6-keto prostaglandin F1alpha liberation and renin activity. These results indicate that exogenous prostacyclin promotes renin release, and suggest that renin release is not to be modulated by A23187-induced prostacyclin synthesis in dog renal cortical slices. PMID- 15550788 TI - Anoxia-reoxygenation stimulates collagen type-I and MMP-1 expression in cardiac fibroblasts: modulation by the PPAR-gamma ligand pioglitazone. AB - BACKGROUND: Cardiac remodeling after ischemic injury is a major cause of heart failure. In this process, fibroblast growth and collagen synthesis and degradation play a critical role. Recent studies indicate that ligands of the peroxisome proliferator-activated receptors-gamma (PPAR-gamma) alter cardiac remodeling during chronic ischemia. This study was designed to investigate if the PPAR-gamma ligand pioglitazone would modulate fibroblast growth and collagen type I synthesis (and expression) in cardiac fibroblasts exposed to anoxia reoxygenation (A-R). METHODS AND RESULTS: Cardiac fibroblasts were exposed to anoxia (95% N2/5% CO2) and then reoxygenation (95% air/5% CO2). A-R increased fibroblast growth (MTT assay) as well as collagen type-I synthesis (H-proline incorporation) and protein expression (Western analysis). Concurrently, there was a parallel increase in the expression of matrix metalloproteinase-1 (MMP-1) in fibroblasts. Pretreatment of cardiac fibroblasts with pioglitazone (10 M) reduced all these effects of A-R. Further, A-R stimulated intracellular reactive oxygen species (ROS) generation and activated the redox-sensitive transcription factor NF-kappaB (both P < 0.05). Both these phenomena were inhibited by pretreatment of cells with pioglitazone. CONCLUSION: Thus, it appears that A-R stimulates fibroblast cell growth, collagen type-I synthesis, and MMP-1 expression in cardiac fibroblasts, most likely a result of ROS generation. Inhibition of ROS generation and induction of NF-kappaB in cardiac fibroblasts during A-R may be a mechanism of action of pioglitazone. PMID- 15550789 TI - Effects of eugenol, an essential oil, on the mechanical and electrical activities of cardiac muscle. AB - Eugenol (EUG) acts as a calcium antagonist but effects on the contractile proteins could also occur. We investigated inotropic effects of EUG in rat left ventricular papillary muscles, measuring isometric force, time variables, and post rest potentiation and EUG actions on the effects of Ca2+ (0.62 to 2.5 mM) and isoproterenol (5 ng/ml), on myosin ATPase activity and on the calcium currents in single ventricular myocytes. EUG reduced tension and time variables without altering the sarcoplasmic reticulum activity increasing post-pause relative potentiation. Isoproterenol and Ca2+ counteract these negative inotropic effects. Tetanic tension diminished, but not the myosin ATPase activity suggesting an isolated sarcolemmal effect. EUG 0.1 mM decreased the Ca2+ current amplitude in the entire potential range tested and 0.5 mM almost completely blocked this inward current. Results suggested that EUG depresses force without affecting the contractile machinery and its action is the only dependent blockade of the calcium inward current. PMID- 15550790 TI - Oxidative stress and apoptosis in cardiomyocyte induced by high-dose alcohol. AB - Binge drinking of alcohol causes cardiac dysfunction in some people. The mechanism remains unclear. This study was designed to investigate high doses of alcohol-induced oxidative stress and apoptosis in cardiomyocytes and protective effects of antioxidants. Cardiomyocytes isolated from 1- to 2-day-old Sprague Dawley rats were treated with ethanol at doses of 0 mM, 50 mM, 100 mM, and 200 mM for 24 hours. Vitamin E (1 mM) and vitamin C (0.2 mM) were added to medium 1 hour before addition of ethanol. Results showed typical apoptosis: chromatin condensation, membrane blebbing, shrinkage, and cytoplasm condensation. Apoptosis is concentration-dependent in the range of 0 to 100 mM ethanol (apoptosis rates were respectively 0.68%, 2.03%, and 9.66% at ethanol concentration of 0 mM, 50 mM, and 100 mM). Necrotic cells became greatly increased in the 200 mM ethanol treated group. Intracellular production of reactive oxygen intermediates increased as mitochondrial membrane potential decreased after ethanol treatment. Cytochrome c was found to be greater in the cytosol of the ethanol-treated groups. Activity of caspase-3 was higher in ethanol-treated groups (P < 0.05). Both vitamin E and vitamin C inhibited oxidative stress and myocyte apoptosis in ethanol-treated groups (P < 0.05). In conclusion, our data indicated that acute high-dose ethanol treatment primarily induces cardiomyocyte apoptosis at concentration up to 100 mM while necrosis is predominate at 200 mM. The underlying mechanism appears to involve mitochondrial damage via an increase in oxidative stress and releasing cytochrome c, which activates caspases that initiate chromatin fragmentation and apoptosis. Antioxidants, to a large extent, inhibit oxidative stress and apoptosis induced by ethanol. PMID- 15550791 TI - Levosimendan in off-pump coronary artery bypass: a four-times masked controlled study. AB - We tested the hypothesis that levosimendan produced beneficial hemodynamic effects during and after off-pump coronary artery bypass grafting in patients with good preoperative left ventricular function. Levosimendan at low dose (12 microg/kg), high dose (24 microg/kg), or placebo were administered in thirty-one patients in a randomized and four-times masked controlled study. Heart rate was not significantly different between experimental groups. Significant increases in cardiac output and left ventricular ejection fraction occurred after high-dose (P < 0.001; P = 0.006) and low-dose levosimendan (P = 0.001; P = 0.002). Both doses of levosimendan produced significant increased stroke volume and decreased systemic vascular resistance. Mean arterial pressure, pulmonary capillary wedge pressure, and left ventricular end-systolic volume were not significantly different between groups. The low-dose levosimendan produced better hemodynamic response than high-dose and may be preferable in patients undergoing off-pump coronary artery bypass grafting. PMID- 15550792 TI - Left atrial diameter is a simple indicator of a deficiency in atrial natriuretic peptide secretion in patients with mitral stenosis: efficacy of postoperative supplementation with synthetic human alpha-atrial natriuretic peptide. AB - OBJECTIVE: With regard to a deficiency in atrial natriuretic peptide (ANP) secretion, the relationship between plasma ANP and left atrial diameter measured by echocardiography was examined and the efficacy of postoperative supplementation was evaluated. METHODS: (1) Ninety-six patients with mitral valve disease from 1997 to 2002 (M:F = 65:31, mean-age 65.3 +/- 8.9 years) were studied for relationship analyses. (2) Twenty-six patients with mitral stenosis and left atrial diameter > or = 60 mm undergoing mitral valve replacement (M:F = 17:9, mean-age 67.4 +/- 7.5 years) were randomly allocated to one of two groups; ANP treated group (n = 13, 0.05 microg/kg/min of synthetic human alpha-ANP was postoperatively administered) and Control group (n = 13). RESULTS: (1) There were significant positive correlations between left atrial diameter and plasma renin activity (r = 0.690, P < 0.01) and between left atrial diameter and plasma aldosterone (r = 0.757, P < 0.01). The maximum value of plasma ANP was 249.5 pg/mL accompanied with 56.2 mm of left atrial diameter in 29 patients suffering from mitral stenosis. There was a significant negative correlation between left atrial diameter and ANP in patients with left atrial diameter > 56.2 mm (r = - 0.725, P < 0.0001), whereas there was a significant positive correlation in patients with left atrial diameter < or = 56.2 mm (r = 0.529, P = 0.0066). (2) At 24 hours after operation, the ANP-treated group showed significantly lower plasma renin-activity (9.2 +/- 3.3 versus 36.2 +/- 7.4 ng/mL/h) and aldosterone (113.6 +/- 36.9 versus 473.8 +/- 95.8 pg/mL) than the Control group. CONCLUSIONS: Left atrial diameter can be a simple and useful indicator of a deficiency in endogenous ANP secretion in patients with mitral stenosis, and postoperative ANP supplementation contributes to suppressing further activation of renin angiotensin system during the immediate postoperative period. PMID- 15550793 TI - The use of angiotensin receptor blockers in the treatment of chronic heart failure. AB - Angiotensin receptor blockers (ARBs) have a pharmacological role in the treatment of heart failure through their blockade of the effects of angiotensin II. ARBs, however, lack the potential benefits of inhibiting the breakdown of bradykinin that is seen with ACE-Is. Historically, the medical literature assessing ARBs in the treatment of chronic heart failure have been short in duration and primarily focused on surrogate markers of disease severity. Recent, well-designed clinical trials have shed new light on the potential roles of ARBs in the treatment of chronic heart failure and their effects on mortality in this patient population. In comparison to captopril, losartan has been shown to have similar benefits in cardiovascular mortality and morbidity. In patients with systolic dysfunction who are intolerant to ACE-Is, candesartan has been shown to reduce cardiovascular mortality and hospital admissions for heart failure. In combination with ACE-Is, candesartan and valsartan have been shown to improve heart failure morbidity and, with candesartan, reduced cardiovascular mortality in those with systolic dysfunction. These 2 trials show conflicting mortality information regarding the use of triple therapy with ACE-Is, ARBs, and beta-blockers for systolic dysfunction. In patients with heart failure but preserved systolic dysfunction, candesartan showed no effects on mortality and only modest effects on morbidity. PMID- 15550794 TI - Treatment of depression in primary care: where we are, where we can go. PMID- 15550795 TI - Clinician screening and treatment of alcohol, drug, and mental problems in primary care: results from healthcare for communities. AB - OBJECTIVE: We sought to estimate national rates of screening and treatment of alcohol, drug, and mental (ADM) problems in primary care. DESIGN: This was a cross-sectional survey administered from 1997 to 1998. PARTICIPANTS: Our study included a nationally representative household probability sample of 7301 primary care patients. MEASUREMENT: We used patient self-reports from a telephone survey to estimate rates of screening and treatment of common ADM problems, to examine the types of screening and treatment received, and to investigate adherence with treatment recommendations. Covariates included measures of ADM conditions, physical health, and sociodemographic indicators. RESULTS: Among adult primary care patients, 38.6% (95% confidence intervals [CI] 37.2-40.0) reported clinician screening for an ADM problem. Alcohol or drug screening occurred more frequently (28.3%; 95% CI 27.0-29.6) than screening for depression and anxiety (21.2%; 95% CI 20.1-22.2). Among those screened, 30.1% (95% CI; 27.8-32.4) reported ADM treatment in primary care. Medications (16.4%; 95% CI 14.3-18.5) and counseling (18.2%; 95% CI 16.1-20.3) were the most common treatments. Rates of screening were higher among individuals with ADM disorders, the young and middle aged, and the college educated. Treatment rates were higher among individuals with ADM disorders. CONCLUSIONS: Substantial effort is expended screening and treating common ADM problems in primary care, and these efforts are targeted towards those with ADM disorders. However, only about half of individuals with an ADM disorder report being screened, and among this group, about 60% report receiving any treatment. PMID- 15550796 TI - Receipt of diabetes services by insured adults with and without claims for mental disorders. AB - OBJECTIVES: We sought to determine whether receipt of the American Diabetes Association's recommended clinical services was similar among insured subjects with and without mental disorders during the period of 1996 to 2001. RESEARCH DESIGN: Our study was a retrospective analysis of Blue Cross/Blue Shield of Iowa administrative claims data, 1996-2001. SUBJECTS: We studied 26,020 adults with diabetes; 6,627 (25%) had a coexisting mental disorder. MEASURES: Service receipt included hemoglobin A1c (HbA1c) testing, dilated eye examination, cholesterol measurement, and urine protein testing. We used Cox regression to calculate hazard ratios (HRs) for service receipt after adjusting for demographic, disease, and utilization factors. RESULTS: Mental disorder subjects were more likely to be younger, women, urban residents, have diabetes complications and comorbidity, and to have increased healthcare utilization. Although they received more services (mean, 2.6) than subjects without mental disorders (mean, 2.3), they were less likely to have received a HbA1c test (HR 0.92; 99.9% confidence interval [CI] 0.87-0.97) and a cholesterol measurement (HR 0.92; 99.9% CI 0.86-0.98). Receipt of a dilated eye examination (HR 0.96; 99.9% CI 0.89-1.04) and urine protein test (HR 0.98; 99.9% CI 0.92-1.04) was similar. Service receipt varied by specific mental disorder categorization. Few subjects (< 6%) strictly adhered to the guidelines of the American Diabetes Association. CONCLUSIONS: Receipt of clinical preventive services for both populations was suboptimal. Importantly, subjects with mental disorders were more likely to have diabetic complications, even when controlling for utilization of healthcare services, possibly because of poorer receipt of HbA1c testing. Persons with mental disorders should be more aggressively educated about blood sugar control, given the high rate of complications in this population. Medical care directed at persons with comorbid medical and psychiatric disorders may be beneficial. PMID- 15550797 TI - Effects of pharmaceutical promotion on adherence to the treatment guidelines for depression. AB - OBJECTIVES: We sought to examine the impact of direct-to-consumer advertising (DTCA) and pharmaceutical promotion to physicians on the likelihood that (1) an individual diagnosed with depression received antidepressant medication and that (2) antidepressant medication was used for the appropriate duration. RESEARCH DESIGN AND SUBJECTS: A quasiexperimental design was used to examine treatment patterns of 30,621 depressed individuals whose insurance claims were included in the MarketScan database from 1997 through 2000. The main explanatory variables were spending on DTCA, detailing to physicians, and free samples for 6 antidepressant medications. RESULTS: Individuals diagnosed with depression during periods when class-level antidepressant DTCA spending was highest (cumulative spending more than US 18.5 million dollars) had 32% higher relative odds of initiating medication therapy compared with those diagnosed during periods when DTCA spending was lowest (P < 0.0001). Free samples of medications dispensed to physicians had no effect on odds of initiating antidepressant use. Class-level DTCA spending on antidepressants had a small positive effect on the duration of antidepressant use, whereas DTCA spending for the specific medication taken by an individual had no effect on treatment duration. Detailing spending at the class or product level had no significant effect on duration of treatment with an antidepressant medication. CONCLUSIONS: Our results suggest that DTCA of antidepressants was associated with an increase in the number of people diagnosed with depression who initiated medication therapy. DTCA was associated with a small increase in the number of individuals treated with antidepressants who received the appropriate duration of therapy. Promotion to physicians was not associated with either the initiation of treatment with an antidepressant or with the duration of therapy. PMID- 15550798 TI - Do the effects of quality improvement for depression care differ for men and women? Results of a group-level randomized controlled trial. AB - OBJECTIVE: We sought to examine whether a quality improvement (QI) program for depression care is effective for both men and women and whether their responses differed. DESIGN: We instituted a group-level, randomized, controlled trial in 46 primary care practices within 6 managed care organizations. Clinics were randomized to usual care or to 1 of 2 QI programs that supported QI teams, provider training, nurse assessment and patient education, and resources to support medication management (QI-Meds) or psychotherapy (QI-Therapy). PATIENTS: There were 1299 primary care patients who screened positive for depression and completed at least one questionnaire during the course of 24 months. OUTCOME MEASURES: Outcomes were probable depression, mental health-related quality of life (HRQOL), work status, use of any antidepressant or psychotherapy, and probable unmet need, which was defined as having probable depression but not receiving probable appropriate care. RESULTS: Women were more likely to receive depression care than men over time, regardless of intervention status. The effect of QI-Meds on probable unmet need was delayed for men, and the magnitude of the effect was significantly greater for men than for women; therefore, this intervention reduced differences in probable unmet need between men and women. QI reduced the likelihood of probable depression equally for men and women. QI Therapy had a greater impact on mental HRQOL and work status for men than for women. QI-Meds improved these outcomes for women. CONCLUSIONS: To affect both quality and outcomes of care for men and women while reducing gender differences, QI programs may need to facilitate access to both medication management and effective psychotherapy for depression. PMID- 15550799 TI - Monitoring depression treatment outcomes with the patient health questionnaire-9. AB - BACKGROUND: Although effective treatment of depressed patients requires regular follow-up contacts and symptom monitoring, an efficient method for assessing treatment outcome is lacking. We investigated responsiveness to treatment, reproducibility, and minimal clinically important difference of the Patient Health Questionnaire-9 (PHQ-9), a standard instrument for diagnosing depression in primary care. METHODS: This study included 434 intervention subjects from the IMPACT study, a multisite treatment trial of late-life depression (63% female, mean age 71 years). Changes in PHQ-9 scores over the course of time were evaluated with respect to change scores of the SCL-20 depression scale as well as 2 independent structured diagnostic interviews for depression during a 6-month period. Test-retest reliability and minimal clinically important difference were assessed in 2 subgroups of patients who completed the PHQ-9 twice exactly 7 days apart. RESULTS: The PHQ-9 responsiveness as measured by effect size was significantly greater than the SCL-20 at 3 months (-1.3 versus -0.9) and equivalent at 6 months (-1.3 versus -1.2). With respect to structured diagnostic interviews, both the PHQ-9 and the SCL-20 change scores accurately discriminated patients with persistent major depression, partial remission, and full remission. Test-retest reliability of the PHQ-9 was excellent, and its minimal clinically important difference for individual change, estimated as 2 standard errors of measurement, was 5 points on the 0 to 27 point PHQ-9 scale. CONCLUSIONS: Well validated as a diagnostic measure, the PHQ-9 has now proven to be a responsive and reliable measure of depression treatment outcomes. Its responsiveness to treatment coupled with its brevity makes the PHQ-9 an attractive tool for gauging response to treatment in individual patient care as well as in clinical research. PMID- 15550800 TI - The effect of improving primary care depression management on employee absenteeism and productivity. A randomized trial. AB - OBJECTIVE: To test whether an intervention to improve primary care depression management significantly improves productivity at work and absenteeism over 2 years. SETTING AND SUBJECTS: Twelve community primary care practices recruiting depressed primary care patients identified in a previsit screening. RESEARCH DESIGN: Practices were stratified by depression treatment patterns before randomization to enhanced or usual care. After delivering brief training, enhanced care clinicians provided improved depression management over 24 months. The research team evaluated productivity and absenteeism at baseline, 6, 12, 18, and 24 months in 326 patients who reported full-or part-time work at one or more completed waves. RESULTS: Employed patients in the enhanced care condition reported 6.1% greater productivity and 22.8% less absenteeism over 2 years. Consistent with its impact on depression severity and emotional role functioning, intervention effects were more observable in consistently employed subjects where the intervention improved productivity by 8.2% over 2 years at an estimated annual value of US 1982 dollars per depressed full-time equivalent and reduced absenteeism by 28.4% or 12.3 days over 2 years at an estimated annual value of US 619 dollars per depressed full-time equivalent. CONCLUSIONS: This trial, which is the first to our knowledge to demonstrate that improving the quality of care for any chronic disease has positive consequences for productivity and absenteeism, encourages formal cost-benefit research to assess the potential return-on investment employers of stable workforces can realize from using their purchasing power to encourage better depression treatment for their employees. PMID- 15550801 TI - Disease management programs for depression: a systematic review and meta-analysis of randomized controlled trials. AB - BACKGROUND: Substantial deficits in the care of depression make the provision of new evidence-based care models a matter of increasing importance. So far, disease management programs (DMPs) have not been systematically assessed. OBJECTIVE: This study was a systematic review and meta-analysis of randomized controlled trials investigating the effectiveness of DMP for depression as compared with usual primary care. METHODS: Criteria for study selection were depression as main diagnosis in adults, the intervention DMP (evidence-based guidelines, patient/provider education, collaborative care, reminder systems, and monitoring), and trial quality A/B (Cochrane Collaboration guidelines) rated by 2 observers. Measurement instruments had to be published in peer-reviewed journals and filled out by the participants, their relations, or independent raters. Meta analyses were conducted by using dichotomous outcomes within forest plots. Tests of heterogeneity, sensitivity analyses, and funnel plots were performed. Economic evaluations were descriptively summarized. RESULTS: DMP had a significant effect on depression severity, with a relative risk of 0.75 (95% confidence interval 0.70-0.81) in a homogeneous dataset of 10 high-quality trials. It was robust in all sensitivity analyses (evidence level 1A). Funnel plot symmetry indicated a low probability of publication bias. Patient satisfaction and adherence to the treatment regimen improved significantly, but only in heterogeneous models. The costs per quality adjusted life year ranged between US 9,051 dollars and US 49,500 dollars. CONCLUSION: DMP significantly enhance the quality of care for depression. Costs are within the range of other widely accepted public health improvements. Future research should focus on the effect of long-term interventions, and the compatibility with health care systems other than managed care driven ones. PMID- 15550802 TI - Quality of depression care in a population-based sample of patients with diabetes and major depression. AB - OBJECTIVES: Major depression occurs in approximately 11% to 15% of patients with diabetes and is associated with poor glycemic control and adverse medical outcomes. This study examined the rates and predictors of recognition of depression among primary care patients with diabetes and comorbid major depression and the quality of depression care provided during a 12-month period. METHODS: This study used automated utilization, pharmacy, and laboratory data from a health maintenance organization to describe the rate of recognition of depression and quality of care provided for patients with major depression and diabetes in the 12-month period before diagnosis. Major depression was diagnosed based on the Patient Health Questionnaire (PHQ-9) that was included in a mail survey sent to 9063 patients on the Group Health diabetes registry from 9 primary care clinics. RESULTS: Approximately 51% of patients with major depression and diabetes were recognized as depressed by the health care system. Women were more likely to be recognized (odds ratio [OR] 1.58, 95% confidence interval [CI 1.26 1.97]), as were those with dysthymia (OR 3.44, 95% CI 2.08-5.72), panic attacks (OR 1.55, 95% CI 1.19-2.19), patients with more than 7 primary care visits (OR 1.42, 95% CI 1.06-1.91) and patients reporting poor health (OR 1.62, 95% CI 1.04 2.53). Of the 51% of patients with major depression who were recognized, 43% received 1 or more antidepressant prescriptions but only 6.7% received 4 or more psychotherapy sessions during a 12-month period. DISCUSSION: There were large gaps in both recognition and quality of depression care provided to patients with major depression and diabetes within a health maintenance organization system. PMID- 15550803 TI - The Revised Behavior and Symptom Identification Scale (BASIS-R): reliability and validity. AB - BACKGROUND: To assess outcomes of health services, providers need brief, responsive, reliable, and valid measures that can be implemented in clinical settings with minimal cost and burden. The Behavior and Symptom Identification Scale (BASIS-32) is a self-report measure developed in 1984 to assess mental health treatment outcomes. During the past 3 years, multiple methods were used to revise the instrument to improve reliability, validity, and applicability to diverse groups of mental health service recipients. OBJECTIVE: The objective of this study was to field test the revised instrument, make further changes based on analysis of the field test data, and assess reliability and validity of the final version (BASIS-24). METHODS: A field test was implemented at 27 treatment sites across the United States. A total of 2656 inpatients and 3222 outpatients participated. Factor analytic methods, classic test theory, and item response theory modeling were used to select the most discriminating, nonredundant items for inclusion in the final version of the instrument and to assess its reliability and validity. Item response theory modeling was used to score the instrument. RESULTS: The final instrument includes 24 items assessing 6 domains: depression/ functioning, interpersonal relationships, self-harm, emotional lability, psychosis, and substance abuse. Test-retest and internal consistency reliability were acceptable. Tests of construct and discriminant validity supported the instrument's ability to discriminate groups expected to differ in mental health status, and its correlation with other measures of mental health. CONCLUSIONS: Analyses of the BASIS-24 supported its reliability and validity for assessing mental health status from the patient's perspective. PMID- 15550804 TI - Use of psychotropic medications for patients with office visits who receive a diagnosis of panic disorder. AB - OBJECTIVE: We sought to examine trends in the medication management of panic disorder from a nationally representative sample of visits to psychiatrists and primary care physicians in office-based practice. METHOD: We analyzed physician reported data from the 1992-1995 and 1996-1999 National Ambulatory Medical Care Survey, focusing on medication management of panic disorder by primary care physicians and psychiatrists. Rates for which doctors prescribed medication for patients with a diagnosis of panic disorder were compared over the course of time to identify changes in medication management between physician groups. We used logistic regression models to identify predictors of medication prescription, adjusting for the presence of other covariates. RESULTS: Both physician groups prescribed a psychotropic medication to a majority of patients for panic disorder in each time period. In 1992-1995, primary care physicians were less likely than psychiatrists to prescribe psychotropic medications. This difference was not evident in 1996-1999. The prescribing of benzodiazepines was common in both physician groups and time periods. Overall rates of psychotropic prescriptions by physician groups converged over time, although no single drug class accounted individually for the convergence. CONCLUSION: In recent years, changes in the medication management of patients with panic disorder by primary care physicians have resulted in greater similarity with the pharmacologic treatment of panic disorder provided in psychiatric practice. PMID- 15550805 TI - Achieving consensus across diverse stakeholders on quality measures for mental healthcare. AB - OBJECTIVE: Quality-improvement efforts are hindered by a lack of consensus on meaningful and feasible measures of care. The objective of this study was to develop a core set of quality measures for mental health and substance-related care that are meaningful to stakeholders, feasible to implement, and broadly representative of diverse dimensions of the mental health system. METHOD: A 12 member panel of stakeholders from national organizations evaluated 116 process measures in a 2-stage modified Delphi consensus development process. Drawing on a conceptual framework and literature review, panelists rated each measure on 7 domains using a 9-point scale (1 = best). Measures were then mapped to a framework of system dimensions to identify a core set with the highest ratings for system characteristics within each dimension. RESULTS: Twenty-eight measures were identified assessing treatment (12), access (2), assessment (2), continuity (4), coordination (2), prevention (1), and safety (5). Overall, mean ratings for meaningfulness were: clinical importance 2.29; perceived gap between actual and optimal care 2.59; association between improved performance and outcome 2.61. For feasibility, mean ratings were clarity of specifications 3.39; acceptability of data collection burden 4.77; and adequacy of case mix adjustment 4.20. The measures address a range of treatment modalities, clinical settings, diagnostic categories, vulnerable populations, and other dimensions of mental healthcare. CONCLUSIONS: A structured consensus process identified a core set of quality measures that are meaningful and feasible to multiple stakeholders, as well as broadly representative of the mental healthcare system. By yielding quantitative assessments of meaningfulness, feasibility and degree of consensus among stakeholders, these results can inform ongoing national efforts to adopt common quality measures for mental healthcare. PMID- 15550806 TI - No quick fix. PMID- 15550807 TI - Leadership: do you have the motivation? PMID- 15550808 TI - Healthy body, healthy mind: discovering a new me. PMID- 15550809 TI - Obesity surgery. PMID- 15550810 TI - Anesthesia for bariatric surgery. PMID- 15550811 TI - Nutritional considerations in bariatric surgery. PMID- 15550812 TI - Body contouring after massive weight loss. AB - As the results of bariatric surgery become increasingly successful, the demand for dy contouring following significant weight loss is rapidly growing. Persons who have lost weight are often unpleasantly surprised with the hanging folds of tissue and loose skin that may be present. The following article is a review of the surgical treatment currently available to provide a more pleasing body contour after successful weight loss. PMID- 15550814 TI - Intraoperative positioning and care of the obese patient. AB - The perioperative nurse involved in the intraoperative care of the obese patient is faced with numerous issues and challenges. As a growing number of these patients present for medical care, the nurse must consider the special positioning needs for surgery and the equipment needed to promote the safest environment for the patient. This article addresses positioning considerations for the obese patient and special equipment needs and selection in the operating room. PMID- 15550815 TI - An ethical wrinkle on the face of therapy claims. PMID- 15550816 TI - Bariatric surgery and reimbursement. PMID- 15550818 TI - Moving forward together. PMID- 15550819 TI - Full-thickness skin grafting: a procedural review. PMID- 15550821 TI - One nurse's experience in obtaining Joint Commission accreditation for office based surgery centers. AB - Obtaining and maintaining Joint Commission accreditation for the office-based facility can be a positive learning experience for nurses, doctors, and ancillary staff. The advantages of such accreditation to the organization and the patients under its care are extensive. This article outlines why offices undergo accreditation and provides insight into how it can be achieved. PMID- 15550822 TI - State-by-state laws for insurance coverage required for post-mastectomy breast reconstruction. PMID- 15550824 TI - Lip augmentation. PMID- 15550825 TI - Assessing learning preferences. PMID- 15550826 TI - Should incentives be used to increase organ donation? PMID- 15550828 TI - Using noncompete agreements. PMID- 15550829 TI - Practicing in "uncharted waters". PMID- 15550830 TI - Hypertensive emergencies. PMID- 15550831 TI - Advances in magnetic resonance. PMID- 15550832 TI - Bile resistance of coated transjugular intrahepatic portosystemic shunt stents in a flow-model. AB - PURPOSE: We sought to test the bile resistance of transjugular intrahepatic portosystemic shunt (TIPS) stents with 3 different coatings. MATERIALS AND METHODS: Three stents with different coating materials (monolayer polyethylene terephthalate [PETP], monolayer polytetrafluoroethylene [PTFE], and double layer [PTFE]) were tested in a flow model. After testing the sealing of the system with isotonic saline solution, fresh human bile was circulated. Constant pressure was 50 cm H2O. Bile resistance of the stent membranes was analyzed. RESULTS: Two of the 3 stents proved completely resistant to water. Only the PETP stent was resistant to bile. The PTFE-coated stents were not bile resistant. CONCLUSION: The bile resistance of coated TIPS stents and, thus, the dependency of TIPS shunt patency is called into question. The stent with the reported superior patency rates does not show experimental bile resistance. PMID- 15550833 TI - Dark lumen magnetic resonance colonography in a rodent polyp model: initial experience and demonstration of feasibility. AB - PURPOSE: We sought to assess dark lumen magnetic resonance (MR) colonography for the detection of colon polyps in a rodent model with histology as the gold standard. MATERIAL AND METHODS: Fourteen male Wistar rats were subjected to carcinogenic N-methyl-N'-nitro-N-nitrosoguanidine at the age of 4 months to induce colon neoplasms. MR imaging was performed after a time interval of 1 year. Preparation and data acquisition was performed with the animals under full anesthesia. After a body-warm saline enema images were acquired on a clinical 1.5 T whole-body MR system using a standard extremity coil. Plain and contrast enhanced (0.3 mmol/kg; Gd-DOTA; Dotarem, Guerbet, France) 3-dimensional T1 weighted gradient recall echo images were acquired. Two radiologists analyzed the MR data sets in consensus for lesion depiction. Contrast uptake in colonic wall and polyps was quantitatively assessed by signal-to-noise ratio and contrast-to noise ratio measurements and compared using a Wilcoxon-Mann-Whitney U test with statistical significance at a P value < 0.05. Finally, all animals were killed, and the MR imaging results were compared with pathologic findings. Sensitivity and specificity were calculated. RESULTS: By pathology, a total of 15 polyps were found in 9 of 14 rats. MR colonography detected 13 of 15 polyps measuring between 4 and 11 mm (mean 7 +/- 0.6 mm) in 8 of 9 animals, resulting in a sensitivity and specificity of 0.87 and 1.0, respectively. Compared with the precontrast data, all polyps showed a statistically significant increase in signal-to-noise ratio (78.2 +/- 6.3 to 167.4 +/- 17.7) and contrast-to-noise ratio (45.4 +/- 5.2 to 124.6 +/- 11.2). CONCLUSION: MR colonography with a dark colon lumen and a bright, contrast-enhanced colon wall appears well suited for the detection of colonic lesions in a rodent model. PMID- 15550834 TI - Quantification of functional mitral valve regurgitation in patients with congestive heart failure: comparison of electron-beam computed tomography with cardiac catheterization. AB - OBJECTIVES: We sought to determine the agreement between electron-beam computed tomography (CT) and cardiac catheterization for the quantification of mitral regurgitation and to evaluate their association with echocardiographic assessment. MATERIAL AND METHODS: Fifty patients with congestive heart failure were examined both by electron-beam CT and catheterization to calculate mitral regurgitation volume and fraction based on the difference between the left ventricular stroke and aortic flow volume. The severity of regurgitation was also compared with visual assessment by echocardiography (grade, 0-4+). RESULTS: The mean values for the mitral regurgitation volume and fraction did not differ significantly between electron-beam CT and catheterization (mean differences: 0.2 mL/m2 and -0.9%, P > 0.05 each, limits of agreement: -14.0 to 14.4 mL/m2 and 26.3 to 24.5%, respectively) and showed a good correlation (r = 0.79 and r = 0.76, respectively; P < 0.05 each). Good levels of correlation were observed between echocardiographic severity grading and quantitative measurements of regurgitation volume and fraction, which were somewhat better between echocardiography and electron-beam CT (rS = 0.78 and rS = 0.84, respectively; P < 0.05 each) than between echocardiography and catheterization (rS = 0.72 and rS = 0.81, respectively; P < 0.05 each). CONCLUSION: Our results suggest that electron beam CT allows for quantification of mitral valve regurgitation with similar accuracy as cardiac catheterization. Measurements with both modalities correlated well with the results of echocardiographic assessment. PMID- 15550835 TI - Normal brain maturation characterized with age-related T2 relaxation times: an attempt to develop a quantitative imaging measure for clinical use. AB - OBJECTIVES: We studied age-related changes in T2 relaxation times from the normal maturating human brain under routine clinical MR examination conditions. MATERIALS AND METHODS: In 70 healthy subjects aged between 3 weeks and 39 years, T2 maps of the brain in which the intensity of each pixel corresponded to T2 relaxation times were generated based on magnetic resonance imaging data collected with a triple spin echo sequence. T2 relaxation times in white matter (WM) and gray matter (GM) were measured in 6 distinctive regions of interest of the T2 maps. The age dependence of the T2 values was mathematically simulated using a biexponential function. RESULTS: T2 values were largest at the age of 3 weeks (maximum: approximately 400 milliseconds for WM and 200 milliseconds for GM) and decreased continuously with increasing age, faster in the first few months and slower thereafter, until values achieved between 95 and 110 milliseconds for WM and 88 and 95 milliseconds for GM in adults. The relationship between T2 values and age could be well simulated using a biexponential function (R > 0.92). CONCLUSIONS: T2 relaxation time correlates well with the progress of brain maturation. The used biexponential function reflects the dynamic development of myelination in newborns and young children as well as the maturation of myelination during adolescence and could be used to develop a "normal" reference for neuroradiological diagnoses. PMID- 15550836 TI - Automated short-axis cardiac magnetic resonance image acquisitions: accuracy of left ventricular dimension measurements in normal subjects and patients. AB - RATIONALE AND OBJECTIVE: This study investigates the use of an automated observer independent planning system for short-axis cardiovascular magnetic resonance (MR) acquisitions in the clinical environment. The capacity of the automated method to produce accurate measurements of left ventricular dimensions and function was quantitatively assessed in normal subjects and patients. METHODS: Fourteen healthy volunteers and 8 patients underwent cardiovascular MR (CMR) acquisitions for ventricular function assessment. Short-axis datasets of the left ventricle (LV) were acquired in 2 ways: manually planned and generated in an automatic fashion. End-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF), and left ventricular mass (LVM) were derived from the 2 datasets. The agreement between the manual and automatic planning methods was assessed. RESULTS: The mean differences between the manual and automated CMR planning methods for the normal subjects and patients were 5.89 mL and 1.93 mL (EDV), 1.14 mL and -0.41 mL (ESV), 0.81% and 0.89% (EF), and 4.35 g and 3.88 g (LVM), respectively. There was no significant difference in ESV and EF. LVM significantly differed in both groups, whereas EDV was significantly different in the normal subjects and insignificantly different in the patients. The variability coefficients were 2.8 and 3.59 (EDV), 3.3 and 5.03 (ESV), 1.79 and 2.65 (EF), and 4.36 and 2.27 (LVM) for the normal subjects and patients, respectively. The mean angular deviation of the LV axes turned out to be 8.58 +/- 5.76 degrees for the normal subjects and 8.35 +/- 5.15 degrees for the patients. CONCLUSIONS: Automated CMR planning method can provide accurate measurements of LV dimensions in normal subjects and patients, and therefore, can be used in the clinical environment for functional assessment of the human cardiovascular system. PMID- 15550837 TI - T2-weighted turbo spin-echo images, maximum-intensity projections, and three dimensional volume-rendering for delineation of pathologies and anatomic details of the inner ear. AB - PURPOSE: The aim of the current study was to compare 3-dimensional T2-weighted turbo spin-echo (TSE) axial slices, maximum-intensity projections (MIPs), and volume-rendered images with respect to the delineation of pathologic findings and anatomic structures of the inner ear. Each of the compared image types was available at 2 different resolutions. MATERIALS AND METHODS: In 64 patients, 3 dimensional T2-TSE images were acquired with a matrix of 128 x 100 and 230 x 230 (field-of-view 90 x 90 mm). Direct axial images, MIPs, and volume-rendered images from the 2 datasets were evaluated in terms of the delineation of pathologic findings and anatomic structures. Analysis was performed by 2 radiologists in consensus. RESULTS: Though axial images acquired with a matrix of 230 x 230 showed more anatomic details than images acquired with a matrix of 128 x 100, pathologic findings (33 in total) were assessable with the same confidence on images of either resolution. Pathologies completely surrounded by fluid were best assessed on axial slices. The delineation of pathologies not embedded in a fluid was almost equally good in axial slices, MIPs, or 3-dimensional volume-rendered images. CONCLUSION: Reading the axial images cannot be replaced by viewing postprocessed reconstructions for initial diagnosis, because only some of the pathologies can be delineated on MIPs and volume reconstructions. However, reconstructions can be valuable for assessing the extent of pathologies and their spatial relation to the anatomic structures. PMID- 15550838 TI - Towards a single-sequence neurologic magnetic resonance imaging examination: multiple-contrast images from an IR TrueFISP experiment. AB - OBJECTIVE: The objective of this study was to reconstruct images bearing multiple contrasts from a single sequence magnetic resonance imaging (MRI) experiment. MATERIALS AND METHODS: Using a segmented IR-TrueFISP imaging sequence, the signal recovery after inversion and alpha/2 preparation was sampled in 6 volunteers. These images were used to generate T1, T2, and spin-density maps, allowing construction of images with multiple contrasts, including T1-, T2-, spin-density weighted, and also FLAIR contrast. Traditionally acquired images bearing the corresponding contrast were obtained for comparison. Regression analysis was performed to compare the synthetic and traditionally acquired images for the whole brain and a region of interest in the occipital region. RESULTS: The synthetic images closely reproduced the contrast from the "standard" examination. Using regression analysis, the obtained image signal intensities for the calculated images compare favorably (P <0.0001-<<0.000001) with images acquired using multiple sequences. CONCLUSIONS: Perfectly registered images with any desired contrast based on T1, T2, and spin density, along with underlying quantitative maps, can be obtained using a single IR-TrueFISP sequence. PMID- 15550839 TI - Breast cancer and the NSABP-B32 sentinel node trial. PMID- 15550840 TI - Clinical considerations in breast cancer sentinel lymph node mapping: a Moffitt review. AB - Lymphatic mapping redefined the clinical significance of axillary lymph nodes in the treatment of breast cancer. Current literature supports the concept that any patient diagnosed with invasive breast cancer should in fact have their sentinel lymph nodes evaluated. However, there are many cases of " special situations in lymphatic mapping " . These special situations mark an important point that should be considered: When and who should undergo lymphatic mapping? A summary of these considerations/cases will be the focus of this report. PMID- 15550841 TI - Sentinel lymph node biopsy after neoadjuvant chemotherapy. AB - BACKGROUND: We surveyed single-center and multi-center studies pertaining to sentinel lymph node biopsy (SLNB) after neoadjuvant chemotherapy to compare the results with those of our current study to evaluate the feasibility and accuracy of SLNB after neoadjuvant chemotherapy. METHODS: From October 2001 to July 2003, 80 patients who had neoadjuvant chemotherapy underwent curative surgery and axillary lymph node dissection (ALND) after SLNB at the Center for Breast Cancer, National Cancer Center. A MEDLINE search was performed using the keywords breast cancer, sentinel lymph node biopsy, and neoadjuvant chemotherapy. RESULTS: Our results showed that 42 (52.6%) of 80 patients had downstaging of the primary tumor; 9 patients (11.3%) had pathologic complete response (pCR) and 33 (41.3%) had pathologic partial response (pPR). 26 patients (32.5%) showed complete axillary clearance after neoadjuvant chemotherapy. Among them, 5 patients (6.3%) revealed pCR of both the primary tumor and axillary metastasis. SLNB was successful in 61 of 80 patients (76.3%) and there were 3 false negatives, yielding a false negative rate (FNR) of 7.3% (3/41), a negation prediction value (NPV) of 87.0%(20/23), and an accuracy of 95.1% (58/61). Thirteen out of 16 studies retrieved by to MEDLINE pertaining SLNB after neoadjuvant chemotherapy concluded its feasibility and accuracy with a identification rate of 82%-100% and a FNR of 17-100%. CONCLUSION: Most studies, including ours, concluded that SLNB after neoadjuvant chemotherapy is accurate and could be an alternative to ALND. PMID- 15550842 TI - Surgical pathological staging of breast cancer by sentinel lymph node biopsy with special emphasis on the histological work-up of axillary sentinel lymph nodes. AB - Axillary nodal status assessed by traditional histological methods is a proven independent prognostic factor in breast cancer. Sentinel lymph node biopsy is a surgical pathologic staging procedure that not only allows the selective removal of the most likely sites of lymphogenic metastases, but also enables upstaging of breast carcinoma by detecting nodal involvement undetected by standard methods of nodal staging. This review highlights the upstaging potential of sentinel node biopsy. It also suggests that incomplete reporting of the pathological methods may make the comparisons of results from different studies difficult. The article also describes a few methods that have been claimed optimal but are probably not, and it formulates basic considerations for building up a histological protocol that can identify all metastases larger than 2 mm, which are of unquestionable prognostic relevance. These considerations are also useful for the detection of micrometastases. Issues of pathological reporting of sentinel nodal findings are also highlighted, with emphasis on the lack of standardization and on the differentiation of isolated tumor cells from micrometastases. Finally the stepwise building up of our current histology protocol and our experience gained since the introduction of sentinel node biopsy in 1997 is also briefly summarized. PMID- 15550843 TI - Lymphoscintigraphy for the visualization of sentinel lymph nodes and body contour. AB - BACKGROUND: It is important to create clear lymphoscintigraphic images when assessing the sentinel lymph nodes. This clarity needs to reach a level where the sentinel lymph nodes (SLNs) and the body contour are clearly visible. We have developed a simple image processing method using the division of primary and scattered photon counts. METHODS: Twenty patients with breast cancer were enrolled in this study. Manual injection of 150 MBq of Tc-99m tin colloid with small particle size into the peritumoral and subdermal regions was performed. Lymphoscintigraphy using a conventional gamma camera was performed three hours after the injection. Dual energy windows were set from 130 to 150 keV for the primary photons and 70 to 110 keV for the scattered photons. An anterior view of the chest and a lateral view from the affected side were obtained. Primary photon image counts were divided by the scattered photon image counts for each pixel after the addition of some constant counts to each pixel of the acquired image to improve the contrast of the scintigrams. We evaluated the ability to accurately visualize the body contour and the SLNs on the processed image. RESULTS: Image processing time was 20 to 40 seconds for each patient. In every case, the processed image clearly identified the body contour. The processed images allowed the identification of the same number of SLNs as the original images. CONCLUSIONS: This proposed method for image processing is a simple and useful means to clearly visualize both SLNs and body contours. PMID- 15550845 TI - Immunohistochemical assessment for estrogen receptor and progesterone receptor status in breast cancer: analysis for a cut-off point as the predictor for endocrine therapy. AB - BACKGROUND: An immunohistochemical (IHC) method is commonly used for determining estrogen receptor (ER) and progesterone receptor (PR) status in breast cancer. However, the proper cut-off points of IHC have not been established. Cut-off points for ER and PR status as predictive factors for endocrine therapy are needed. METHODS: A total of 249 cases of female breast cancer were enrolled. ER and PR status by IHC were analyzed using the proportion of stained cells and staining intensity by Allred's score. RESULTS: Proportion score (PS) and intensity score (IS) were related to enzyme immunoassay (EIA) titers, for both in ER and PR (p < 0.0001, all). PS correlated with IS in both ER and PR (R = 0.47 and 0.41, respectively). ER status by IHC was related to tumor size and lymph node status, while PR was related to tumor size and menopausal status. In 152 patients who received endocrine therapy with a median follow-up term of 38 months, differences in disease-free survival were most significant using a cut off point of PS 3 which indicated more than 10 % of cells stained positively for both ER and PR (p = 0.0007 and 0.0087, respectively). In addition, combination analysis of ER and PR using this cut-off point revealed a notable prognostic difference. CONCLUSION: A 10 % staining proportion may be an acceptable cut-off point for both ER and PR status by IHC, in terms of predicting response to endocrine therapy in breast cancer. PMID- 15550844 TI - Optimal particle size of radiocolloid for sentinel node identification in breast cancer-electron microscopic study and clinical comparison. AB - BACKGROUND: Sentinel node (SN) biopsy using a radiotracer is easy to learn, reproducible, and performed by the majority of breast cancer surgeons. However, this technique raises the question: what is the optimal particle size of colloid? METHODS: Patients were injected with conventional tin colloid (median particle diameter: 700 nm) and the predominant particle size of radiocolloid retained in the SN was measured using electron microscopy. This showed a narrow distribution of colloidal size (100-150 nm). Patients were then injected with modified tin colloid having a median particle size of 100 nm. A clinical comparison was performed between conventional particle size or reduced particle size tin colloid and the SN biopsy success rate, the number of SNs, and the colloidal uptake in SNs. RESULTS: A total of 118 patients were injected with the conventional tin colloid (group I) and 124 patients with the smaller particle colloid (group II). The identification rate and the number of SN in both groups were almost equal, and the patients with low-uptake SNs were not significantly less in group I(p = 0.55). However, in the subgroup of patients 60 years of age or more, group II had significantly more SNs than group I(1.4 vs 1.9; p = 0.03) and low-uptake SNs were significantly less common than in group I(p = 0.02). CONCLUSIONS: The improvement of colloidal uptake in SNs using the smaller particle size tin colloid was confirmed, and this impact was statistically significant in the older population. PMID- 15550846 TI - Is early recognition of radiologically silent brain metastasis from breast cancer beneficial? A retrospective study of 22 cases. AB - Over a period of 10 years, twenty-two patients with T2-3N2Mx breast cancer presented with symptoms of increased intracranial pressure (ICP), but brain CT scan and/or MRI did not reveal any signs of a space occupying lesion. A brain CT scan and an MRI study were performed every 15 days. Ten patients refused this close follow up. Thus, two groups were formed group A (n = 12) included the patients who were close's followed and group B (n = 10) consisted of those patients who were not. Therefore, Group A, being under careful follow-up, initiated radiotherapy were quickly. The median time from the presentation of increased ICP symptoms until the appearance of metastases on CT and/or MRI directly followed by brain irradiation was 48 days (SE = 6.1) for group A and 72 days (SE = 0.7) for group B (p = 0.0085, log-rank test). In group A, median Overall Survival (OS) was 171.0 (SE = 21.5) days, and was 99.0 (SE = 6.3) days (p = 0.014) for group B. Volumetric analysis of the secondary brain lesions at the initiation of radiotherapy showed a total volume of metastatic lesions of 19.5 +/ 13.9 cm3 versus 65.3 +/- 20.7 cm3 for groups A and B, respectively (p < 0.001, Mann-Whitney test). Post-radiotherapy, Karnofsky Performance Status and Visual Analogue Score were also improved for group A versus B (p = 0.002). Group A appeared to benefit from close follow-up since the metastases were detected and irradiation was given sooner compared with group B, thus achieving better performance status and prolonged survival. Radiologically silent brain metastases from breast cancer should not be ignored because timely whole brain irradiation should be the goal of clinicians. Clinical suspicion should lead to close follow up with multiple CT/MRI studies and cerebral spinal fluid cytology until the final diagnosis. PMID- 15550847 TI - Inhibition of bone metastasis from breast cancer with pamidronate resulting in reduction of urinary pyridinoline and deoxypyridinoline in a rat model. AB - BACKGROUND: Breast cancers frequently metastasize to bone, in a process in which osteoclasts play a major role. Bisphosphonate pamidronate, a specific inhibitor of osteoclasts, has been widely used in the treatment of bone metastasis (BM). In this study, using an animal model of BM, we examined the prophylactic and treatment effects of pamidronate against BM and clarified the relationships between BM, pamidronate and bone resorption markers such as urinary pyridinoline and deoxypyridinoline. METHODS: Bone metastases were established by inoculating c SST-2 (spontaneously developed rat mammary adenocarcinoma) cells into the thoracic aorta of 27 rats, which were then divided into three groups of rats: the untreated control group, the pre-treatment group, consisting of rats treated with pamidronate (10 mg/kg) injected subcutaneously a day before tumor inoculation, and the post-treatment group, in which rats were injected with pamidronate a week after tumor inoculation. Three weeks after tumor inoculation, blood and urine samples were collected. The subjects were then sacrificed to harvest the thoracic and lumbar vertebrae for histological examination, consisting of staining with hematoxylin and eosin and tartrate resistant acid phosphatase (TRACP). RESULTS: The incidence of BM was 70.0%, 44.4% and 37.5% in the control, pre-treatment and post-treatment groups, respectively. Although there was no significant difference among the groups, the rate of BM in the treated groups was lower than that of the control group and no bone destruction was observed in treated rats. The TRACP stained specimens revealed that there were numerous osteoclasts contributing to the control group tumor burden. The urinary levels of pyridinoline and deoxypyridinoline were reduced by pamidronate. CONCLUSION: Our results suggest that pamidronate prevents the development of BM and the destruction of bone associated with BM. Maintaining the values of Pyr and Dpyr at low levels with pamidronate might lead to inhibition of the incidence and development of BM. PMID- 15550848 TI - Evaluation of dynamic studies of MR mammography for the diagnosis of intraductal lesions with nipple discharge. AB - OBJECTIVES: We assessed the utility of dynamic magnetic resonance imaging (MRI) in differentiating benign from malignant lesions of the breast and then applied MRI to diagnose intraductal breast tumors with nipple discharge. METHODS: Gadolinium (Gd)-enhanced MR mammography was performed on 74 patients with breast tumors and 8 patients with nipple discharge. RESULTS: The steepest slopes of the contrast medium uptake (S slope) s from time-intensity curves were significantly different between malignant and benign lesions. At S slope threshold of 0.95% /second, malignancy was predicted with a sensitivity and specificity of 75% . Six of 8 cases with nipple discharge were successfully identified by MR ductography by injecting Gd-DTPA into discharging ducts. Among them, 2 non-invasive ductal carcinomas were differentiated from benign lesions by the S slope value. CONCLUSIONS: Dynamic MR mammography is an useful modality for differentiating breast lesions and has potential for evaluating intraductal lesions with nipple discharge. PMID- 15550849 TI - Expression of cdc25A and cdc25B phosphatase in breast carcinoma. AB - BACKGROUND: Previous studies have indicated that cdc25B and cdc25A phosphatase stimulate cell cycle progression and also play a role in malignant transformation of cells. In this study, we investigated the expression of these phosphatases in breast carcinoma to elucidate their significance. METHODS: We immunohistochemically examined cdc25B and cdc25A expression in 46 cases of breast carcinoma. RESULTS: Cdc25B and cdc25A were negative or only equivocally expressed in glandular epithelial cells. In breast carcinoma, 56.5% of cases had high levels of cdc25B expression. Interestingly, the expression of cdc25B was significantly lower in cases with biologically aggressive phenotypes. The cdc25A level was high in 69.6% of cases, but we could not find any relationship between cdc25A expression and clinicopathological parameters. CONCLUSION: These results suggest that cdc25B is important especially in the early phase of breast carcinoma progression. PMID- 15550850 TI - Overexpression of Her2/neu, estrogen and progesterone receptors in invasive micropapillary carcinoma of the breast. AB - BACKGROUND: Invasive micropapillary carcinoma (IMC) of the breast is a rare subtype of breast carcinoma that has an extremely high incidence of lymph node metastasis and poor clinical outcome. Furthermore, there has been interest in the prognostic markers related to the biological characteristics of breast carcinoma, such as the status of Her2/neu, estrogen receptor (ER) and progesterone receptor (PgR). MATERIALS AND METHODS: We reviewed 671 cases of primary breast carcinoma which were surgically resected between 1990 and 2003. Of these, 27 cases of invasive ductal carcinoma of the breast with a pure or partial micropapillary component were reported. Tests for Her2/neu protein, estrogen receptor (ER) and progesterone receptor (PgR) were performed on paraffin sections. The relationship among these markers was analyzed and statistical significance determined by chi square test. CONCLUSION: The rate of Her2/neu protein overexpression, ER and PgR status was quite similar to those of common breast carcinomas. PMID- 15550851 TI - A case of invasive lobular carcinoma of the breast first manifesting with duodenal obstruction. AB - Lobular carcinoma of the breast presents with various clinical manifestations. Lobular carcinoma comprises about one fifth of cases of ductal carcinoma. Intestinal metastasis is dominant in cases of lobular carcinoma. Without a prior diagnosis of lobular carcinoma of the breast, the diagnosis of intestinal metastasis from breast cancer is difficult. A 52-year-old women underwent duodenogastrectomy for duodenal cancer. About 18 months later, she underwent mastectomy for lobular carcinoma of the breast. The duodenal cancer consisted mainly of malignant cells invading the submucosa that were identical to those in the breast. Immunohistochemical assays for estrogen receptor also confirmed breast cancer metastasis. Duodenal obstruction was the first sign of isolated metastatic lobular carcinoma of the breast in this case. The characteristic metastatic pattern of lobular carcinoma, which differs from that of ductal carcinoma, should be kept in mind to ensue proper diagnosis. Preceding breast surgery for lobular carcinoma should prompt investigation based on the typical metastatic pattern. PMID- 15550852 TI - Successful combination therapy with trastuzumab and Paclitaxel for adriamycin- and docetaxel-resistant inflammatory breast cancer. AB - We present a case of adriamycin-and docetaxel-resistant inflammatory breast cancer (IBC) in which partial response was achieved with combination therapy using trastuzumab and paclitaxel. A 48-year old woman noticed a lump in her right breast. She was diagnosed with IBC and the disease was staged as T4d N1 M0, stage III B. The patient was started on neoadjuvant chemotherapy with adriamycin (50 mg/m2) and docetaxel (60 mg/m2) administered every three weeks. Six courses were performed and the response was evaluated as no change. After one month, contralateral breast swelling indicated bilateral IBC. Bilatera1 mastectomy using the Halsted method was performed. The immunohistochemical results of the Hercep Test was strongly positive (3+). After the mastectomy, right pleural effusion appeared, and cytological examination revealed the cells to be classV(adenocarcinoma). To treat the clinically advanced breast cancer, combination therapy with trastuzumab (initially 4 mg/kg followed by two or more cycles of 2 mg/kg) and paclitaxel (80 mg/m2) were given intravenously every week for eight cycles and then every two weeks thereafter. A total of 32 courses of therapy were performed, the pleural effusion completely disappeared and partial response was maintained for a duration of 482 days. The adverse reactions were mild, and it was possible for her to be treated as an outpatient with high quality of life. This report suggests that weekly combination therapy of trastuzumab and paclitaxel was useful for treatment of adriamycin-and docetaxel resistant metastatic breast cancer. PMID- 15550853 TI - Pathological fracture of the femur ten years after successful radiation therapy for metastatic breast cancer. AB - We describe a case involving a 75-year-old woman presenting with a femur fracture 10 years after radiation therapy for metastatic breast cancer, which developed in the right femur. The lesion showed complete response with bone healing following radiation therapy; however, the patient sustained a femur fracture ten years later. Histological examination of the specimens obtained from the lesion revealed features of radiation osteonecrosis, but there was no histological evidence of tumor. To our knowledge, there has been no reported case of pathological fracture ten years after radiation therapy from radiation osteonecrosis rather than progression of the metastatic lesion. Late complications of radiation therapy should be considered with care, even when metastatic lesions demonstrate complete response to treatment. PMID- 15550854 TI - Sensitization of chemotherapy by anti-HER. AB - This article reviews the current state of efforts targeting human epidermal growth factor receptor-2 (erbB2/HER2)in breast cancer therapy. The results of recently conducted clinical studies with trastuzumab and several other compounds are presented. Trastuzumab, a humanised monoclonal antibody(mAb)directed against the extracellular domain of HER2, has been shown to be active against HER2 overexpressing metastatic breast cancer, either as a single agent or when used in combination with chemotherapy. In preclinical models, trastuzumab has shown additive and even synergistic anti-tumor activity with the chemotherapeutic agents. In a large, randomised, phase III trial, the combination of trastuzumab and chemotherapy was shown to improve the response rate and survival in patients with metastatic breast cancer. The high incidence of cardiotoxicity seen with the combination of trastuzumab plus anthracycline drugs prompted several clinical studies combining trastuzumab with other chemotherapeutic agents, including taxanes, vinorelbine and platinum salts. This article summarises the available data on trastuzumab-based combination chemotherapies and novel drugs targeting HER2 and signal transduction molecules for the treatment of breast cancer. PMID- 15550855 TI - Capecitabine plus docetaxel combination chemotherapy for metastatic breast cancer. AB - Doxifluridine(5'-DFUR)is converted to its metabolite 5-FU by the enzyme thymidine phosphorylase(TP). TP is expressed significantly higher in tumor tissue than in normal tissue. Capecitabine(N4-pentoxylcarbonyl -5'-deoxy-5-fluorocytidine)is a pro-drug of 5'-DFUR and a novel fluoropyrimidine carbamate that is converted to 5 FU preferentially in tumor tissue through a three-step enzymatic cascade. Expression of TP in tumor tissue may clinically predict efficacy of capecitabine. Induction of TP activity has brought about enhancement of capecitabine efficacy by taxanes in human cancer xenografts. In addition, a phase III study directly comparison docetaxel monotherapy and docetaxel plus capecitabine has been conducted for metastatic breast cancer patients who had received anthracyclines. The overall response rate of the combination group was 42%(n=255), and that of the monotherapy group was 30%(n=256)(p=0.006). The primary endpoints were time to disease progression, and time to treatment failure, and these parameters were superior in the combination arm than in the single arm, suggesting that capecitabine sensitization by docetaxel might be a new approach to breast cancer treatment. PMID- 15550856 TI - Molecular targets for potentiation of radiation-induced cell killing. AB - Molecular target-based drugs have been emerging as a cancer treatment. Clinical trials using the combined approach of radiation therapy and molecular target based drugs have been performed to evaluate the feasibility of this approach, and improve the response of tumors to radiation. To achieve maximum radiotherapeutic gain, understanding of the interaction of radiation and drugs are indispensable. Preclinical data have already demonstrated synergistic enhancement of radiation induced cell killing by several molecular target-based drugs. Among these, the effect of drugs that target receptor tyrosine kinase and its signal transduction pathways on radiosensitivity has been intensively investigated. In this review, established and potential molecular targets for potentiation of radiation-induced cell killing are summarized, and preclinical data regarding investigations of new molecular targets for radiosensitization will be introduced. In addition, the results and toxicities of clinical trials using combined radiation therapy and molecular target-based drugs are summarized. PMID- 15550857 TI - Present role and future perspectives of the evaluation of the effect of primary chemotherapy by breast imaging. AB - Recently, primary chemotherapy has been used not only for locally advanced breast cancers but also operable cases in an adjuvant setting. Therefore, many randomized clinical trials for primary chemotherapy have been coducted all over the world, with the result that more than one-third of the cases have become nonpalpable (clinical CR). On the contrary, accurate determination of the surgical margin has become more difficult. If some cancer remains as a nonpalpable lesion, it must be detected. Breast MRI has been assuming an important role in the assessment of the extent of cancer and does so more accurately than conventional modalities such as mammography and ultrasonography. Image-guided biopsy or mapping is promising and should be developed as soon as possible. In this article, some promising modalities for the detection of residual lesions after primary chemotherapy are extensively reviewed. PMID- 15550858 TI - Novel sensitizing agents: potential contribution of COX-2 inhibitor for endocrine therapy of breast cancer. AB - Enhancement of the therapeutic effect of conventional drugs is currently an active treatment strategy for breast cancer, as shown in the clinical application of trastuzumab with chemotherapeutic agents, which prolonged survival even for metastatic disease. Cyclo-oxygenase 2(COX-2)inhibitors, which are chemoprevention agents for familial polyposis coli, are now contributing to this strategy in combination with chemotherapeutic and endocrine drugs. As an endocrine application, overexpression of COX-2 contributes to increased expression of aromatase in the breast tumor. In addition, it is also known to promote rich micro-vessels within the tumor through up-regulation of prostaglandin E2(PGE2), which can induce vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)in cancer cells, and can directly modulate endothelial cell proliferation. Since both rich vasculature and accelerated estrogen synthesis are thought to contribute to unfavorable conditions for the response to endocrine therapy, inhibiting COX-2 with COX-2 inhibitors is a promising strategy to potentiate endocrine therapy. PMID- 15550859 TI - Significance of histopathological evaluation in primary therapy for breast cancer -recent trends in primary modality with pathological complete response (pCR) as endpoint. AB - In recent years, primary therapy has been used to improve the prognosis of patients with locally advanced breast cancer and to expand the indication for breast conserving treatment for patients with a relatively early stage of breast cancer. In addition, the therapeutic efficacy of primary therapy has been evaluated on the basis of pathological findings and pathological complete response (pCR)is considered to be a main target of primary therapy. The results of NSABP protocol B-18 and B-27, and the Aberdeen trials confirmed the prognostic significance of pCR in primary therapy and indicated the significance of minute pathological assessment. However, the criteria of pathological response is not yet universal, but the evaluation of the main invasive tumor, intraductal component and the regional lymph nodes, is thought to be necessary, shown by the "Histopathological Criteria for Assessment of Therapeutic Response in Breast Cancer" compiled by the Japanese Breast Cancer Society. Among these criteria, there exist methodological variations as to the evaluation of residual disease of intraductal carcinoma, thus some controversies exist. The presence of intraductal component might be negligible with regard to prognosis, but might be an important risk factor for local recurrence after breast conserving therapy. In the future, participation by the pathologist in the field of primary therapy for breast cancer will be a matter of course in most clinical studies. PMID- 15550860 TI - The evolution of primary chemotherapy in breast cancer treatment. AB - Primary chemotherapy may be thought of as having evolved over three generations. First-generation primary chemotherapy followed trials which proved that the prognosis of breast cancer patients receiving primary chemotherapy was the same as that of patients receiving post-operative chemotherapy. Second-generation primary chemotherapy followed trials which demonstrated that regimens containing anthracyclines and taxanes yielded complete pathological response rates (pCR)of over 20%. Third-generation primary chemotherapy followed trials which demonstrated that inclusion of newer agents, such as trastuzumab and capetitabine, in combination with taxanes could yield pathological CR rates of over 30%. In third-generation primary chemotherapy, predictive factors, especially those analyzed by DNA array analysis, play a key role. Other issues, including the relevance of sentinel node biopsy after primary chemotherapy and the importance of marking the site of the primary tumor before primary chemotherapy in cases showing cCR, are highlighted. This article deals with the history of the evolution of primary chemotherapy, along with a discussion of some relevant issues. PMID- 15550861 TI - What caused the decline in breast cancer mortality in the United Kingdom? AB - BACKGROUND: The marked improvement in breast cancer mortality in the United Kingdom (UK) has attracted worldwide interest. To understand the details of this phenomenon the morbidity and mortality rates of breast cancer in the UK during the past 30 years were analyzed. METHODS: The official publications and statistical data were available and downloaded at the official UK Web sites of the National Health Service, National Statistics and Cancer Research Organization. Parts of the data were obtained directly from the National Cancer Intelligence Centre of UK. RESULTS: After the beginning of breast screening the morbidity of the subject age group soared up to the level of an age group that was 10 years older. Noninvasive carcinoma was often discovered in the subject age group after the beginning of breast screening and constituted 8.3%of all cases. The mortality rates, however, began to decrease before the start of screening and fell clearly even in age groups other than that of the study group and particularly in elderly women. CONCLUSIONS: Not only the prevalence of breast screening but also the increase of early stage cancer in all age groups have probably resulted from improvements in diagnostic technology and heightened social interest and awareness. In addition, the development of adjuvant endocrine therapy must have influenced the evident improvement in mortality of breast cancer in the UK. PMID- 15550862 TI - Prognostic implication of p53 protein expression in relation to nuclear pleomorphism and the MIB-1 counts in breast cancer. AB - BACKGROUND: A close correlation of the p53 protein expression to nuclear pleomorphism and proliferative activity in breast cancer has been reported. The prognostic implications of p53 protein expression, however, in relation to nuclear pleomorphism and proliferative activity in breast cancer remain controversial. PATIENTS AND METHODS: Nuclear pleomorphism and immunohistochemical reactivity for p53 protein and MIB-1 were evaluated on formalin-fixed paraffin stored sections from 250 patients with breast cancer for whom the median follow up duration was 6.4 years. RESULTS: p53 protein expression was positive in 66 (26.4%) of 250 cases. Nuclear pleomorphism was grade I or II in 169 (67.6%) cases and grade III in 81(32.4%)cases. The MIB-1 counts were more than 10% in 102 (40.8%) cases and less than 10% in 148 (59.2%) cases. There was a close correlation between p53 protein expression and nuclear pleomorphism (p<0.0001) and between p53 protein expression and MIB-1 counts (p<0.0001). Univariate analyses showed the 66 cases with positive p53 protein expression to have a significantly (p=0.0284) worse disease free survival (DFS) than the 184 cases with negative p53 protein expression. A multivariate analysis, however, on the variables including all of p53 protein expression, nuclear pleomorphism and MIB-1 counts indicated the MIB-1 counts (p=0.0041) as well as the lymph node status to be independently significant factors for DFS, while neither p53 protein expression nor nuclear pleomorphism were independently significant factors for DFS. CONCLUSION: The present study demonstrated that the p53 protein expression, nuclear pleomorphism and MIB-1 counts all demonstrated prognostic significance for breast cancer, while the most significant prognostic indicator among these three biological parameters was the MIB-1 counts. PMID- 15550863 TI - Clinical and pathologic features of invasive micropapillary carcinoma. AB - BACKGROUND: Invasive micropapillary carcinoma(IMC)of the breast is a rare subtype of breast carcinoma that has an extremely high incidence of lymph node metastases and poor clinical outcome. This histological subtype of breast carcinoma has remained unclear due to the rarity of cases. Many questions exist on the clinicopathological significance of this subtype, especially regarding prognosis. METHODS: We reviewed all 671 cases of primary breast carcinoma that were surgically resected at our institute between 1990 and 2004. Of these, 27 cases of invasive ductal carcinoma of the breast with a pure or partial micropapillary component were reported. The cases were analyzed using various parameters, including age at presentation, tumor size, tumor grade, presence of lymphatic invasion, and axially lymph node status. RESULTS: The patients' age at presentation ranged from 31 to 74 years (mean 52.4 years). Tumor size ranged from 0.7 to 10 cm (mean 4 cm). 88.9% (24 of 27) of the cases had lymphatic invasion. Of cases who underwent with axillary dissection, 66.6% (18 of 27) had positive lymph nodes. Clinical follow up data were available for 17 cases with IMC for 1 to 72 months. Among these, 10 patients died from breast carcinoma within 5 years. CONCLUSION: While our series is too small to make conclusions about the behavior of IMC, the difference in 6-year survival rate between the patients with IMC and those with breast carcinoma in general was statistically significant. Recognition of this distinctive and aggressive variant of infiltrating carcinoma is important because of its poor prognosis and high incidence of lymph node metastases. PMID- 15550864 TI - Follow-up of cases with false-negative pathologic sentinel nodes in breast cancer. AB - BACKGROUND: The clinical practice of sentinel lymph node biopsy for breast cancer patients started in 1999 in our hospital, to obviate unnecessary axillary lymph node dissection. The present study examines the pathological false-negative cases on intraoperative sentinel lymph node investigations and evaluates their outcomes. METHODS: The subjects consisted of 183 cases with clinically node negative breast cancer who had undergone sentinel node biopsy. When the sentinel node was noted to contain malignant cells intraoperatively, a complete axillary lymph node dissection was performed subsequently. The patients with tumor free sentinel nodes underwent no further axillary surgery. The pathological false negative cases in this series were defined as patients with lymph node involvement which was revealed postoperatively, despite negative intraoperative sentinel node examinations. After these surgeries and/or adjuvant therapies, interval clinical evaluations were performed for all patients. RESULTS: Intraoperative diagnosis of the sentinel node was 96.2% accurate compared with the results of permanent sections. There were six pathological false-negative cases, a false-negative rate of 4.1%, all of which had only micrometastasis. Five cases received systemic adjuvant therapy and have been disease-free, however, one patient who refused further therapy developed infraclavicular lymph node metastasis two years after surgery. CONCLUSIONS: In the management of the patients with postoperatively revealed sentinel node micrometastasis, systemic adjuvant therapies might reduce local relapse without secondary lymph node dissection. PMID- 15550865 TI - Breast cancer in Japanese men: does sex affect prognosis? AB - BACKGROUND: Breast cancer has received much less investigative attention in Asian men than in Caucasian men. We examined whether the prognosis of Japanese men with breast cancer differs from that of Japanese women with the disease. METHODS: The clinicopathological features of 14 Japanese men with breast cancer were reviewed and age- and stage-matched case-control analysis of these men and 140 female patients was performed. RESULTS: Disease-free survival (p=0.94) and overall survival (p=0.62) did not differ significantly between the sexes. Five-year disease-free survival was 77% for the men and 75% for the women, and the 5-year overall survival was 92% for the men and 86% for the women. The disease recurred in 2 men but none died of breast cancer, although 3 died of other causes during the median follow-up period of 7 years. There were no significant differences in p53 mutation (p=0.20) or erbB-2 oncoprotein overexpression (p=0.33) between the men and women studied. CONCLUSION: Survival rates of Japanese male and female breast cancer patients are similar when age and stage of the disease are taken into consideration. However, comorbid disease mortality is likely the major contributor to clinical outcome in Japanese male breast cancer. PMID- 15550866 TI - Clinical experience of weekly paclitaxel-based treatment as preoperative chemotherapy for patients with primary breast cancer. AB - BACKGROUND: Paclitaxel is an effective agent in the treatment of metastatic breast cancer. The aim of this study was to evaluate the safety and efficacy of weekly paclitaxel-based preoperative chemotherapy in patients with large operable breast cancer. METHODS: Patients initially received paclitaxel as a 3-hour infusion at 175 mg/m2. Three weeks after initial administration, two cycles of three weeks of paclitaxel 80 mg/m2 over a 1 hour infusion followed by a one week break were given. Of 22 patients, 9 had stage II (tumor diameter greater than 3 cm), 4 stage III A, 7 stage III B, and 2 stage IV (with ipsilateral supraclavicular lymph node metastasis) cancer, respectively. RESULTS: Excluding stage IV patients, the overall response rate to paclitaxel chemotherapy was 80%. Four of the 20 patients (20%) showed a clinical complete response (cCR). Two of these showed pathologic complete response and the other 2 had only the ductal component remaining. The primary tumor response and axillary lymph node downstaging following preoperative chemotherapy tended to be related in 16 patients with clinically positive nodes. Breast conserving surgery was performed as a result of downstaging in the 9 stage II patients. Grade 3 neutropenia occurred in one patient when 175 mg/m2 of paclitaxel was administered, but no serious side effects developed during the weekly administration of paclitaxel. CONCLUSION: The use of weekly paclitaxel-based preoperative chemotherapy appears to yield a significant anti-tumor effect without inducing serious drug-related adverse effects. Furthermore, the effectiveness of this treatment appears to result in a higher frequency of breast conserving surgery. PMID- 15550867 TI - A phase II study of S-1 in patients with metastatic breast cancer--a Japanese trial by the S-1 Cooperative Study Group, Breast Cancer Working Group. AB - BACKGROUND: S-1 is a newly developed novel oral dihydrouracil dehydrogenase inhibiting fluoropyrimidine drug consisting of 1 M tegafur (FT), 0.4 M 5-chloro 2, 4-dihydroxypyrimidine (gimeracil), and 1 M potassium oxonate (oteracil), with efficient antitumor activity and low gastrointestinal toxicity which is widely used in Japan against advanced gastric, head and neck cancers. We investigated its clinical efficacy against metastatic breast cancer. METHODS: A non-blind phase II study was carried out to evaluate the efficacy and toxicity in metastatic breast cancer patients. Patients with measurable metastasis foci (n=111) were enrolled, and 108 patients were regarded as eligible. S-1 was administered orally at a standard dose of 80 mg/m2/day b.i.d. One course consisted of 28 consecutive days of administration followed by a 14-day rest, and courses were repeated up to six times. RESULTS: Among the eligible patients, 10 had a complete response and 35 had a partial response, with an overall response rate (CR+PR) of 41.7% (95% confidence interval: CI, 32.3-51.5%). The incidences of toxicity (> or =grade 3) were neutropenia 9.1%, anemia 0.9%, anorexia 3.6%, stomatitis 1.8%, nausea/vomiting 1.8%, diarrhea 0.9%, and fatigue 2.7%, however no treatment-related deaths were observed. The median survival time was 872 days (95% CI, 572-1,110 days). There was no difference in response rate or toxicity between the under 65-year-old group and the older group. CONCLUSION: S-1 was demonstrated to have high efficacy with low gastrointestinal toxicity even in older patients and will be a promising new chemotherapy drug for metastatic breast cancer. PMID- 15550868 TI - A case of serum CEA disappearance curve after resection of breast carcinoma. AB - Carcinoembryonic antigen (CEA) elimination kinetics after tumor resection were measured in a case of breast cancer. A 45-year-old woman with a left breast carcinoma underwent surgery after neoadjuvant chemotherapy. The serum CEA level before surgery was 34.3 ng/ml. After sequential monitoring of serum CEA levels, postoperative serum CEA elimination kinetics were calculated using non-linear least square analysis with the fitting equation C(t)=(C0-Cp)exp(-kt)+Cp, where C(t) was the postoperative CEA level, t was the number days after surgery, C0 was the CEA level at postoperative time zero, Cp was the CEA at plateau, and k was the rate constant of elimination. Cp was calculated as 6.9 ng/ml, which was above the cut-off level and indicated residual malignancy. After adjuvant chemotherapy, CEA normalized to 1.8 ng/ml. In breast cancer patients with high preoperative serum CEA levels, our analytical method for CEA elimination might be useful for the detection of residual malignancies. PMID- 15550869 TI - A case of chondromatous tumor of the breast. AB - This report describes an exceedingly rare case of a benign cartilage-containing breast tumor that developed in the right breast of a 52-year-old woman. She found the mass on self examination. Physical examination revealed a 1.5 x 1.4 cm, firm, smooth and mobile lump in the lower medial quadrant close to the nipple of the right breast. Mammography revealed a slightly indistinct margined, oval-shaped, and high density nodule without microcalcifications. On ultrasonography, the lesion was a hypoechoic, oval-shaped mass with an echogenic spot. The border was slightly rough. Fine needle aspiration cytology revealed some giant cells and necrotic tissue. Excisional biopsy was then performed. Histopathologically, the lesion consisted of islands of mature hyaline cartilage with intervening strands of fibrous stroma. Mammary lobules and ducts were lacking within the mass. Fat and muscular components were not present. Therefore chondromatous tumor of the breast was diagnosed. PMID- 15550870 TI - A case of breast cholesterol granuloma accompanied by cancer. AB - Cholesterol granuloma of the breast is a very rare benign disease with clinical and imaging features that are often indistinguishable from cancer preoperatively. We report a case of breast cholesterol granuloma accompanied by cancer. The patient was a 78-year-old woman who complained of a lump in her right breast. Mammography and ultrasonography showed a well-circumscribed mass. Fine needle aspiration cytology showed many cholesterol crystals and inflammatory cells without malignancy. With a diagnosis of cholesterol granuloma, tumor extirpation was performed. Histopathologic examination revealed cholesterol granuloma together with breast cancer, and additional partial mastectomy was subsequently performed. It is noted that breast cholesterol granuloma could be accompanied by cancer. PMID- 15550871 TI - Syringomatous adenoma of the nipple: a case report. AB - Syringomatous adenoma of the nipple is a rare disease. To our knowledge, only 23 cases have been reported in the literature. We present the case of a 36-year-old lactating woman with syringomatous adenoma of the nipple, in whom local excision of the tumor was performed. Histologically, the tumor consisted of tubules, ductules and epithelial cell strands, and most of the proliferating ducts showed a characteristic teardrop or comma-shaped appearance. Careful monitoring to detect local recurrence is considered necessary, because syringomatous adenoma of the nipple, while being benign, is a locally invasive tumor. PMID- 15550872 TI - A case of breast cancer with predominant cartilaginous and osseous components. AB - Primary breast cancers with cartilaginous and osseous components are quite rare. We recently treated a 51-year-old woman suffering from primary breast cancer with predominant cartilaginous and osseous components, adjacent to a component of intraductal carcinoma. We discuss the management of this tumor with a review of the literature. PMID- 15550873 TI - [Risk of infection and prostheses: time out or a red flag?]. PMID- 15550874 TI - [Gynecology-obstetrics: one and indivisible]. PMID- 15550875 TI - [Materno-foetal dialogue and human embryo implantation: some evolving concepts]. AB - Multiple immunogical mechanisms allow fetal allograft tolerance. In this review, we first describe the maternal and embryological side in order to expose the dangers for the embryo enabling the development of materno-fetal strategies that will allow fetal survival and growth. PMID- 15550876 TI - [Changing attitudes on the surgical treatment of urogenital prolapse: birth of the tension-free vaginal mesh]. AB - OBJECTIVES: To describe, in view of the new standardization, the technique for urogenital prolapse repair using a one-piece synthetic mesh. MATERIALS AND METHODS: The history and development steps through which the Tension free Vaginal Mesh (TVM) technique emerged are described. The use of a mesh was prompted by the 20-30% recurrence rate associated with conventional repair techniques. Selection of the type of mesh is discussed. Non-absorbable synthetic meshes have shown their usefulness in visceral surgery. A list of materials along with their respective advantages and inconveniences is reviewed and particular emphasis is put on both the tolerance and erosion issues, the latter being specific to the vaginal route. The TVM Group selected a one-thread polypropylene mesh, Prolene Soft, which seemed the most appropriate for the transvaginal approach of prolapse surgical repair. The prosthesis and its design rationale are described. Full details are given on the consecutive intervention steps and underlying concepts. RESULTS: The relevant literature is scarce and there is a lack of methodologically sound studies validating the materials and techniques used. After completion of a first step of technique refinement and feasibility assessment involving about 300 surgical interventions, the authors initiated a prospective multicenter study. Clinical outcome assessments using feasibility, complications, and efficacy endpoints will be published after twelve months, three years, and five years of follow-up. CONCLUSION: Fruitful reasoning led to the development of the TVM technique of complete surgical repair of genital prolapse, which uses a synthetic materiel carefully selected after several tests. All surgeons can apply this technique after a short training period. PMID- 15550878 TI - [Laparoscopic colorectal resection for endometriosis: preliminary results]. AB - OBJECTIVE: Colorectal endometriosis is source of chronic pelvic pain greatly affecting quality-of-life. Colorectal resection is indicated after failure of medical treatment. Few data are available on complications and functional results after laparoscopic colorectal resection for endometriosis. Therefore, the aims of this prospective study were to evaluate the feasibility, peri-operative complications and functional results of laparoscopic colorectal resection for endometriosis. MATERIALS AND METHODS: From March 2001 to March 2003, 32 consecutive women with clinically-suspected colorectal endometriosis confirmed by MR imaging and rectal endoscopic sonography were included in this prospective study. RESULTS: Conversion to open surgery was required for four of the 32 women (12.5%). Mean operating time was 6 hours (range 4 to 13). Associated surgical procedures were: adhesiolysis (n=24), ureteral lysis (n=19), ovarian cystectomy (n=11), and hysterectomy (n=4). Mean blood loss was 2.4 g/dl (range: 0 to 8.6). Blood transfusion was required in 6 women including two who underwent laparoconversion. Two rectovaginal fistulae (6.3%) occurred requiring a colostomy. Urinary retention was noted in 6 women (15.6%). CONCLUSION: Laparoscopic colorectal resection for endometriosis is feasible and is associated with a significant improvement of symptoms. However, the benefit of this procedure has to be weighed against the high morbidity. PMID- 15550877 TI - [Primary breast sarcoma. A retrospective study of 42 patients treated at the Bergonie Institute during a 32-year period]. AB - OBJECTIVE: To evaluate the experience of a single cancer center with unusual tumors. To analyze Primary breast sarcomas (PBS). To investigate treatment and prognostic factors influencing overall survival (OS) and disease-free survival (DFS). PATIENTS AND METHODS: Retrospective study of a series of 42 patients. We reviewed the clinical records and pathology slides of 42 women with PBS treated in our institution between 1970 and 2002. Log-rank tests were used to determine OS and DFS. RESULTS: The median age at diagnosis was 56.9 years (24-81 years). Surgery was part of the therapeutic strategy in all the patients. Patients with angiosarcoma and those with malignant cystosarcoma constituted distinct populations. The 10-year OS and DFS rates were 53% and 55% for angiosarcoma patients and 89% and 100% for cystosarcoma patients (p=0.009 and 0.01 respectively). CONCLUSION: Careful preoperative multidisciplinary assessment is required before making the decision to treat. Mastectomy is generally indicated. Axillary lymph node dissection is not indicated. PMID- 15550879 TI - [Pregnancy and delivery in adolescents: characteristics and profile of 311 cases]. AB - OBJECTIVE: Adolescent pregnancy is considered to be a high-risk situation in a large part of the world. The aim of this study is to assess the validity of this notion for Moroccan teenagers. MATERIAL AND METHODS: From January 1999 to December 2000, there were 311 teenagers who gave birth in the "Les orangers" maternity hospital in Rabat, Morocco. We compared these adolescents with 155 women aged more than 18 years who delivered during the same period. RESULTS: We were unable to demonstrate a high risk for complications among our adolescent population. The only statistically significant differences found between the two groups concerned use of contraception (22.4% for teenagers vs. 57.6%), traditional practices (6.1% vs. 1.93%), frequency of episiotomy (90% vs. 67.14%), instrumental extraction (37.32% vs. 27.14%), lower birth weight in teenagers (3074 g vs. 3199 g) and a higher rate of low birth weight in unmarried adolescents in comparison with married ones (23.3% vs. 9.6%). CONCLUSION: Adolescent pregnancy in Morocco remains well accepted, particularly within the context of marriage. PMID- 15550880 TI - [Pregnancy beyond age 40 in 382 women: a retrospective study in Reunion Island]. AB - OBJECTIVE: To examine obstetrical and neonatal outcomes among women age 40 years and older. MATERIAL AND METHODS: There was a retrospective study including 382 pregnant women at least 40 years of age and 7786 controls aged 20-34, stratified according to parity. Associations between maternal age and pregnancy outcomes were assessed using Pearson's chi(2) test, Fischer exact test, Student test or Mann and Whitney test, as appropriate. RESULTS: Maternal age 40 and over was associated with an increased risk for gestational diabetes (multiparas: OR 3.2, 95%CI 2.2-4.8; grandmultiparas: 2.8, 1.6-5.0), chronic diabetes (multiparas: 3.5, 1.04-10.6), chronic hypertension (multiparas: 3.1, 1.5-6.1; grandmultiparas: 12.1, 3.3-53.2), pregnancy-induced hypertension (nulliparas: 4.6, 1.01-17.3; multiparas: 2.6, 1.3-5.4) and preclampsia (multiparas: 2.9, 1.1-7.2). The risk for cesarean section was 2-fold higher in older multiparas and grandmultiparas. The same was true for the rate of operative vaginal delivery. The rate of fetal chromosomal abnormalities was 7-fold higher in older gravidas. Rates of fetal death were significantly increased in the cohort of older pregnant women (nulliparas: 11.2, 2.9-44.0; multiparas: 3.9, 1.4-10.9). CONCLUSION: Pregnancy at age 40 and over is associated with high rates of obstetrical complications, cesarean sections and operative vaginal deliveries, emphasizing the importance of rigorous antenatal care. PMID- 15550881 TI - [Psychological approach to hyperemis gravidarum]. AB - INTRODUCTION: Hyperemesis gravidarum is a phenomenon that concerns only 1 or 2% of pregnant women who require hospitalization if their anorectic behavior does not cease. The treatment consists in complete isolation, anti-vomit drugs and rehydration therapy. OBJECTS: Isolation has been practiced existed since 1914, and is based on the belief that these women were simulating pain in order to obtain abortion. By isolating them from their family environment, the medical staff hopes this secret desire will be revealed. We tried to evaluate the reality of this analysis. MATERIAL AND METHOD: Two psychological interviews were conducted by a questionnaire addressed to 23 pregnant women at the time of their hospitalization for hyperemesis gravidarum in a level 3 gynecology-obstetric unit. All patients were submitted to the described treatment. RESULTS: Four groups were found. In the first group (n=6) women were submerged by a present and conscious conflict concerning their pregnancy, which found and easy solution. In the second group (n=7), the women had lost a pregnancy a short time before the present one. These patients were suffering from this loss and could not easily accept their new baby. The third group (n=6) concerned women who were afraid of a living a dramatic event that their mother had experienced in the past. This worry, held far from their consciousness, could be recalled during the interviews. In the last group (n=4), the women had passed through major difficulties in their relationship with their mothers during childhood. Becoming a mother themselves, they feared they would reproduce the same attitudes toward their own babies. Within this group, only one woman finally asked for an abortion that was performed. CONCLUSION: It would not appear that the patients have to be isolated for them to reveal their desire for abortion. Conversely, a psychological interview allows better understanding of the ambivalent attitudes of pregnant women and thus can be useful in avoiding systematic abortion. PMID- 15550882 TI - [Obstetrical and neonatal prognosis of monoamniotic twin gestations]. AB - Monoamniotic twin gestations result from a late division of the fertilized ovum with development of the two embryos within a common amnionic sac. We performed a retrospective study in our university hospital on nearly 19 years of clinical activities; we found 16 cases of monoamniotic twins. Obstetrical and neonatal outcomes were analyzed and compared to the data in the literature. The pronosis after 28 weeks of gestation does not appear to be influenced by this type of twin gestation and we demonstrate in this study that vaginal delivery is possible and not deleterious. PMID- 15550883 TI - [Kell alloimmunization in pregnancy]. AB - INTRODUCTION: Kell alloimmunization is a rare disease, although its incidence is the highest after after anti-D alloimmunization. METHODS: We report two recent cases and a review of the literature to describe practical management of Kell alloimmunization in pregnancy. DISCUSSION: When an immunization against the Kell antigen was diagnosed, amniocentesis was performed at 14 weeks gestation to determine the fetal blood group. If the fetus was Kell positive, a first fetal blood sample was drawn at 17 weeks gestation in case of fetal hydrops, and at 20 weeks without fetal hydrops. The diagnosis of anemia led to in utero transfusion. A second fetal blood sample was taken at 8 to 10 days, every two weeks during the second trimester and every three or four weeks during the third trimester. Fetal well-being was assessed with weekly sonography and rates of hemoglobin decline. These measures enable adapting the frequency of fetal blood sampling. PMID- 15550884 TI - [First detection of sentinel node in adenocarcinoma of Bartholin's gland]. AB - We report the first case of detection of sentinel node in a 54 year-old woman presenting an adenocarcinoma of Bartholin's gland. Primary carcinoma of Bartholin's gland is rare and represents 2-7% of vulvar malignant lesions; this could explain the lack of consensus about treatment. The best attitude could be vulvectomy and inguinal lymphadenectomy. Pelvic lymphadenectomy is not required when no pelvic sentinel node is observed or when no metastatic inguinal node can be detected. PMID- 15550885 TI - [Another look at the implications of the DCCT study]. AB - The fundamental role of good metabolic control has been demonstrated in type 1 and type 2 diabetes. Nevertheless, clinicians often wonder why some patients under good metabolic control develop complications while others remain free of such complications, despite a poorly controlled disease. The present study revisited material from the DCCT database, by classifying the 1441 patients as being under good or poor metabolic control if their HbA1c mean level fell in the lower (HbA1c<=6.9%) or upper (HbA1c>/=9.5%) quintile of the overall distribution of mean HbA1c levels observed in the DCCT population. The impact of metabolic control and of other potential factors related to the patient and his/her disease on the development and/or deterioration of complications, in particular diabetic retinopathy and nephropathy, was assessed. Although metabolic control is the major determinant of the risk of developing diabetic retinopathy and nephropathy, the study also emphasizes the significant role of other risk factors, in particularly BMI, disease duration, micro-albuminuria, HbA1c at baseline, gender and age on such complications. It is concluded that early control of the metabolic and clinical status of diabetic patients has major consequences on the evolution of the disease. Nomograms have been proposed to help the clinician in this task. PMID- 15550886 TI - [The quality-of-life questionnaire QLS-H: validation of the French language version of the questionnaire in patients with growth hormone deficiency and collection of reference data in the general population]. AB - The QLS-H(c) (Questions on Life Satisfaction- Hypopituitarism) is new a quality of life (QoL) self-administered questionnaire addressing the complaints of adult patients with growth hormone deficiency. The French version of the QLS-H(c) (16 items) has been psychometrically evaluated during a randomized, open label study comparing two strategies of growth hormone (GH) replacement therapy. Seventy three patients were included and received an 8-month GH replacement therapy. QoL was explored at baseline, 4 and 8 months using the QLS-H(c) questionnaire and the Nottingham Health Profile (NHP) reference scale. Acceptance of the QLS-H(c) was excellent as 92% of the questionnaires were suitable for analysis. All the items demonstrated good selectivity. The homogeneity of the questionnaire was confirmed (Cronbach's alpha, 0.87). The external validity construct was assessed and confirmed using the NHP scores. Sensitivity to change was confirmed. Following an 8-month replacement therapy, the perception of the QoL assessed with the QLS-H(c) questionnaire was significantly improved, irrespective to the treatment strategy. Finally, redundant items of the questionnaire were removed. As a result, the final version of the QLS-H(c) contained 9 items. In a parallel study, reference data of the QLS-H(c) (9 items) were collected from a representative sample of 989 subjects from the French population. With these reference ranges, algorithms to calculate Z scores adjusted for age and gender were developed as a measure for the deviation of patients' scores from those of the general population, and also to evaluate changes along time. In summary, the French version of the quality of life QLS-H(c) questionnaire is a relevant, validated investigational tool for the evaluation and follow-up of an adult patient with growth hormone deficiency. PMID- 15550887 TI - [Induction and reversibility of thyroid proliferative changes in suckling rats given thiocyanate]. AB - Potassium thiocyanate given in the drinking water of pregnant rats led to decreased body weight in their 14-day-old offspring (27%) without altering thyroid weight. Reduction of the suckling rat's body weight could be explained be defective thyroxinemia (38). Plasma FT3 and TSH were unchanged after thiocyanate treatment. The biochemical changes were in agreement with the histological aspects of the hypothyroid animals. The typical pattern was hyperplastic goiter. Colloid volume was reduced compared with controls. Presence of resorbed peripheral vacuoles, a sign of thyroid hyperactivity, was disclosed by a three fold increase in radioiodide (131I) uptake compared with controls. When the antithyroid drug was removed from the mother's milk, the pups'weight increased but did not reach control values. Plasma thyroid hormone levels returned to normal and even exceeded control values in spite of partial recovery of thyroid iodine content when thiocyanate treatment was stopped for ten days. PMID- 15550888 TI - [Oral manifestations of endocrine dysfunction]. AB - Oral manifestations of endocrine dysfunction often may be observed initially by the dentist. Objective manifestations, such as ophtalmos in hyperthyroidism, signs of hypersecretion of GH in acromegaly, are easily recognized. Dentists should have some knowledge of many other diseases in this category that occasionally come in our attention. The present article will discuss the effects of over and under-secretion of each endocrine gland separately, showing its influence on the development and maintenance of the health of the teeth and supporting structures. Diabetes mellitus is the most common endocrinological disease, with an incidence of 3%. Periodontitis risk is three time greater in diabetic patients than in general population and it may worsen the diabetes evolution. Periodontitis in diabetic patients needs an rapid diagnosis and treatment. We also presents the oral aspects of thyroid, parathyroid, suprarenalian, growth hormone and female hormones pathology. The incidence of these troubles is less important, but oral manifestations may reveal an endocrine disfunction. PMID- 15550889 TI - [Gonadal dysgenesis associated with Mayer-Rokitansky-Kuster-Hauser syndrome: a case report]. AB - Gonadal dysgenesis with female phenotype is defined as the absence or insufficient development of the ovaries. Hypogonadism or impuberism are variable, depending on the degree of gonadal development. Mayer-Rokitansky-Kuster-Hauser syndrome is a rare malformative anomaly (1/5000 women) associating uterine and vaginal aplasia with normal ovaries. We report the case of a 19-year-old woman who presented primary amenorrhea and impuberism. Hormone assay revealed hypergonadotrophic hypogonadism. The karyotype was normal, 46XX. Internal genitalia could not be identified on the pelvic ultrasound. Laparoscopy was undertaken and revealed concomitant ovarian dysgenesis and Mayer-Rokitansky Kuster-Hauser syndrome. There were no other morphological malformations. An association between these two conditions is very exceptional and appears to be coincidental, independent of chromosomal anomalies. Hormone substitution therapy remains the only therapeutic option. Hormone substitution is aimed at triggering the development of secondary sexual characters and prevent osteoporosis. There remains the unsolved problem of infertility. PMID- 15550890 TI - [Recurred hydatid cyst of the thyroid: a case report]. AB - A 21-year-old patient underwent surgery in 1987 for hydatid cyst of segment VIII of the liver. Six years later, the patient developed a cold thyroid nodule. During the thyroid operation, needle aspiration and cytology study confirmed the diagnosis of hydatid cyst. Resection of the prominent dome was performed. In 1996, the patient developed cervical recurrence. Pericytectomy was attempted and during the procedure a wound of the subclavian artery required sternoclavicular disarticulation. Hydatid cyst of the thyroid gland is rare, but should be considered in patients living in endemic zones who develop an anterior neck tumor. Cystectomy, isthmolobectomy or sub-total thyroidectomy may be indicated. Resection of the prominent dome may be required in the event of recurrence. PMID- 15550891 TI - [Introduction]. PMID- 15550892 TI - [Element of digestive pathophysiology and fecal analysis]. AB - Fecal analysis includes qualitative and quantitative studies which allows quantification and labelling of numerous pathophysiologic phenomenona. Malabsorption and over-absorption of water and electrolytes give rise to six types of watery diarrheas, and two types of constipations; malabsorption of nutriments and maldigestion of food, give rise to two types of fatty and nitrogenous diarrheas with metabolic consequences. Fecal analysis often discriminates organic from non-organic diseases and brings informations on increase or decrease of caloric losses, to the nutritionist. Microscopic observations which requires a high degree of competence and experience, allows the recognition of malabsorption/maldigestion phenomenona, of fortuitous presence of parasites and a good interpretation of a fecal file. PMID- 15550893 TI - [New fecal markers: recent developments and perspectives]. AB - Fecal occult blood testing is the most widely prescribed screening test for colorectal cancer. Recent development of immunological tests has increased specificity. Fecal DNA analysis opens up a new field for early detection of this widespread neoplasia. Inflammatory bowel disease is another important area where the development of fecal markers provides an interesting alternative to the gold standard but costly and invasive endoscopic investigations with histological analysis of biopsy specimens. Fecal TNFalpha and calprotectin can now be proposed to distinguish organic from non-organic intestinal disease, so select candidates for further investigations, and to assess disease activity. Measurement of fecal elastase provides real progress in screening for exocrine pancreatic insufficiency in patients with malabsorption syndrome. The development of non invasive fecal markers is thus of increasing interest, providing data about the entire gastrointestinal tract useful for screening and individual patient management. PMID- 15550894 TI - [Quality control in coprology]. AB - Quality control in medical laboratories was defined in guidelines for good execution of laboratory analyses issued by the French health authorities in 1994. Application of these guidelines is difficult in coprology because the sample is a complex heterogeneous matrix which varies with disease, surgery, food intake, and treatment. In addition, commercial quality control kits are not available for stool biochemical analyses and a national quality control program has not been established. We thus developed our own fecal quality control technique using pooling lyophylized stool samples. Manual or partially automated methods are used in coprology, leading to a long pre-analysis phase which is not always taken into account in quality control. This implies the need for complementary tools to insure the quality of coprology analyses. For example, semi-quantitative microscopic lipid analysis can be used as an internal standard for a given specimen. Quality assurance also involves a post-analytical phase where results obtained for a given specimen are compared with other available data and interpreted in light of the patient's clinical and therapeutic status. This quality assurance strategy enables accurate reliable results useful for long-term patient management. PMID- 15550895 TI - [Andrea Cesalpino and systemic circulation]. AB - Although since the publication of the De motu Cordis in 1628 the majority of historians have considered William Harvey the discoverer of the blood circulation, a minority of authors have tried to show that the laurels belong instead to Andrea Cesalpino (c.1520-1603). Over the years,an endless number of points and opinions on the two sides of the question have been advanced and debated; the consensus today, however, is that Harvey deserves the credit and that Cesalpino's contribution was negligible. In an attempt to clarify the issue, the author has translated de novo, from the original Latin, all the passages of Cesalpinoconcerning the circulation and has reviewed the criticisms leveled at his ideas on the subject. The results are surprising on two accounts: first, the conclusion seems inescapable that this author, several decades before Harvey, had a clear general understanding of the circulation of the blood;second, the dismissal of Cesalpino's contribution was (and is) based on misunderstandings and misinterpretations.These, in turn, were the result, for the most part, of two factors: a) critics often did not read all the pertinent passages(possibly because his Latin prose is tedious and sometimes unclear); b) some passages were inaccurately translated. As for who discovered the circulation, it depends on how we define "discovery". PMID- 15550896 TI - [Applications of molecular biology to care of patients with malignant hematological disease]. AB - The improvement of the techniques of molecular biology allowed greater performances in the field of detection and characterization of chromosomal abnormalities and/or molecular defects observed in human hematological malignancies. Cytological and immunophenotypical results are reinforced by data obtained with standard and molecular cytogenetic tools and with PCR based techniques. Molecular data are usefull at diagnosis in order to define different types of leukemias and to score patient prognosis. Therefore, these techniques help to choose among various therapeutic options. In post induction treatment period, PCR tools allowed to measure minimal residual disease (MRD) and to distinguish new prognosis factors essential for patient's management. PMID- 15550897 TI - [Intestinal bacterial colonisation and translocation in C3H/HeJ conventional mice fed a unique dose of live or dead Bifidobacterium breve C50]. AB - The efficiency and safety of use of Bifidobacterium breve C50 (BbC50), a potential probiotic, was assessed as regards intestinal microbial colonisation and bacterial translocation. A suspension of BbC50, containing 1-5 to 107-108 live bacteria, was fed to C3H/HeJ mice. The passage of live BbC50 was not demonstrated by culture either in the intestine or extra-intestinal organs. However, mice receiving the highest dose of live bacteria harbored more lactobacilli and less Bacteroides fragilis group in the cecum and colon when compared to control mice. Translocation of lactobacilli observed in the control group was not regulated by Bb50 feeding. Indeed, the spleen was significantly more frequently contaminated in mice fed BbC50, whatever the dose of live bacteria. The kidneys were also significantly more contaminated with lactobacilli in mice fed the highest dose of live Bb50. Moreover, higher dose of live BbC50 was associated with greater number of extra-intestinal contaminated organs. To conclude, BbC50 feeding induced a favorable balance in the mouse intestinal flora and was never found translocating, demonstrating its efficiency and safety of use. However, BbC50 seemed to interfere with the ability of lymphoid organs (e.g. the spleen) to eliminate translocating lactobacilli. PMID- 15550898 TI - [Antimicrobial activity of new 4,4'-bipyridine derivatives: and in vitro study]. AB - The antimicrobial and antifungal activity of some 4,4'-bipyridine derivatives were studied. The diffusimetric gelose surface diffusion method with stainless steel cylinders was used to study bacteria and Sabouraud fields for Candida albicans. Comparative analysis of the results led to the following conclusions. Diquaternary salts of 4,4'-bipyridinium possess a remarkable antimicrobial and antifungal activity. The influence of R1 and R2 substitutes in the para or meta position of the benzoylic radical affects selectivity but does not greatly influence activity. PMID- 15550899 TI - Solitary fibrous tumor of the tentorium cerebelli. Case report. AB - Solitary fibrous tumor (SFT) is a mesenchymal neoplasm that has been recognized to occur almost all along the organism. Since its description in 1996 at the meninges, a total of 59 cases of meningeal SFT have been reported. Different authors have emphasized the difficulties in the differential diagnosis with other more frequent meningeal neoplasms such as meningioma or hemangiopericytoma, as the clinico-radiological characteristics of this lesion seem to be non specific and the morphological features on pathological study may resemble other spindle cell neoplasms. The diffuse and strong reactivity for CD34 and the negativity for EMA and S-100 are data allowing the diagnosis of SFT. We report the case of a 50 year-old woman suffering from headache, in whom MRI study showed a tentorial lesion initially thought to be a meningioma. In spite of morphological similarities with a fibrous meningioma, inmunohistochemical study finally led to the diagnosis of SFT. As occurred in previous cases, the findings in our patient reflect the similarities in clinico-radiological and pathological characteristics between meningeal SFT and other spindle cell meningeal neoplasms, mainly fibrous meningioma. When a clear diagnosis cannot be done based on typical findings on conventional hematoxylin-eosin study, inmunohistochemical study should be performed in meningeal spindle cell lesions to exclude SFT. PMID- 15550900 TI - Spontaneous rupture of spinal dermoid cyst with disseminated lipid droplets in central canal and ventricles. AB - Free fat in the ventricular space is a rare but well recognized complication of ruptured tumour of dermal origin. However, only 1 patient of spontaneous rupture of spinal dermoid tumour with disseminated fat in the central canal and ventricles has been described in the literature. The authors report an extremely rare case of ruptured intraspinal dermoid and passage of free fatty droplets via the patent central canal to the intracranial CSF space. The detailed clinical presentation, radiological findings, and review of the literature are presented. Despite being rarely reported, spinal dermoid cyst can rupture spontaneously, and free fat disseminate into the ventricles, and in extremely rare cases, fat can enter into the central canal. It is underlinerd that a prompt detection, with the help of MRI is essential in cases of spinal dermoid tumour cyst, with sudden deterioration in neurological condition, keeping in mind, the possibility of free fat in the central canal. PMID- 15550901 TI - Neurological deterioration after head trauma in patients with colloid cysts of the 3rd ventricle. Two case histories. AB - The authors report 2 cases of colloid cyst of the 3(rd) ventricle (both patients previously asymptomatic) diagnosed after head trauma. Neurological deterioration due to biventricular hydrocephalus, rendering necessary an urgent bilateral CSF shunting, was observed. The possible relationship between clinical deterioration and head injury in these cases is discussed. PMID- 15550902 TI - Metastasis of breast carcinoma to intracranial meningioma. AB - A patient who had been treated for bilateral breast carcinoma subsequently developed a metastatic breast lesion in a meningioma. Although it is not uncommon for more than one tumor to occur in the same patient, metastases from one tumor into another tumor are rare (''tumor to tumor'' phenomenon). Meningiomas are the most common primary, intracranial tumors to harbor metastases, the majority of which arise from breast and lung carcinomas. Patients with a history of breast cancer and a solitary, intracranial mass with radiological features consistent of meningioma should be evaluated surgically. The lesion may be a primary cerebral malignancy, a metastatic lesion, or a meningioma with or without a metastatic lesion. PMID- 15550903 TI - Isolated, benign, intraorbital schwannoma arising from the supraorbital nerve. Case report and review of the literature. AB - The authors describe the rare case of a 65-year-old man, with absent clinical evidence of von Recklinghausen's disease, harbouring an isolated, benign schwannoma of the right supraorbital nerve. The patient presented a progressive, painless proptosis of the eye for the past 2 years, did not complain of any intra or periorbital pain, and did not experience any form of visual disturbance or field defect. The lesion was completely resected using a fronto-orbitozygomatic approach. The clinicopathological, radiological and surgical features are discussed, and the literature on supraorbital nerve schwannomas reviewed. To the authors' knowledge the fronto-orbitozygomatic approach has not been previously reported for the removal of supraorbital nerve tumors. PMID- 15550904 TI - Postoperative spondylodiscitis from Aspergillus fumigatus in immunocompetent subjects. AB - The authors describe a case of spondylodiscitis from Aspergillus fumigatus which occurred subsequent to surgery for lumbar disc herniation in a non immunodepressed patient. The results obtained by combined medical and surgical treatment are discussed. PMID- 15550905 TI - Sudden death secondary to fulminant intracranial aspegillosis in a healthy teenager after posterior fossa surgery: the role of corticosteroids and prophylactic recommendations. AB - Postoperative complications from corticosteroids in neurosurgical patients are not uncommon. Too often the deleterious immunosuppressive effects of corticosteroids are overlooked in neurosurgery patients and can lead to serious and lethal infections. EXPERIMENTAL DESIGN: case report of a 16-year-old healthy male who presented for elective resection of a recurrent juvile pilocytic astrocytoma of the posterior fossa 4 years after initial resection. SETTING: major University institutional practice. INTERVENTION/RESULTS: a standard suboccipital craniotomy with gross total resection. Postoperatively, the patient suffered from posterior fossa syndrome and diminished gag reflex requiring nasogastric feeds with progressive improvement. While awaiting transfer to a rehabilitation center on postoperative day 12 he suffered a sudden temperature spike followed by neurological decline. A stat computed tomography scan of the brain revealed a diffuse miliary process with severe cerebral edema. Sputum and cerebrospinal fluid cultures identified Aspergillus. Despite immediate therapy to combat the malignant cerebral edema, the patient died within 24 hours of onset of the symptoms. Corticosteroids are used routinely in neurosurgery to combat cerebral edema without much consideration for the immunosuppressive effects. This case demonstrates how the immunosuppressive effects of corticosteroids can lead to a fulminant lethal fungal infection. Neurosurgeons should be aware of the anticatabolic medications now available to combat the deleterious side effects of corticosteroids. PMID- 15550906 TI - Cervical epidural hematoma without fracture in a patient with ankylosing spondylitis. A case report. AB - Ankylosing spondylitis (AS) is an inflammatory disease involving the axial spine. Alterations in vertebral biomechanics leave the spine sensitive to traumas which, though minimal, may cause serious neurological lesions, particularly in long term AS patients with a completely ankylosed spine, who are more prone to suffer spine fractures. A 62-year-old man with a long-term AS suffered a minor trauma resulting in a cervical epidural hematoma from C2 to C7, leading to paraplegia. On the diagnosis of hematoma, he underwent C3-C7 left hemilaminectomies, to remove the hematoma. We could find no cause for the hematoma. The patient's condition improved, and he was eventually able to perform all his activities independently. Even though traumatic spinal epidural hematoma (SHE) of the ankylosed spine may occur in the absence of fracture, it is commonly associated with traumatic fracture or dislocation of the spine, particularly the cervical spine. In all the published series of SEH in AS, we could only find one more case of this pathology at the cervical spine without a fracture. Neurologic recovery can be successful if decompression is performed early. SHE must be considered after trauma to an ankylosed spine when there is neurological deterioration despite the absence of fracture. A good outcome depends on the early diagnosis and surgery. PMID- 15550914 TI - The natriuretic peptides in cardiovascular medicine. AB - Within the last five years, assay systems for measurement of plasma levels of brain natriuretic peptide (BNP) have been approved as a diagnostic aid for heart failure (HF). Similarly, nesiritide, a recombinant form of human BNP, has been approved for the treatment of acutely decompensated HF. Both BNP as a diagnostic test and a therapeutic modality are rapidly becoming integrated into clinical practice. The purpose of this review is to provide a brief overview of the physiology of the natriuretic peptides relevant to their informed clinical use. The current literature regarding the utility of measuring BNP for the diagnosis and management of HF is reviewed and practical recommendations regarding the interpretation of BNP levels are offered. The clinical literature regarding the use of recombinant BNP for the treatment of HF is reviewed, underscoring current gaps in our knowledge regarding the indications for and benefits of this novel agent. PMID- 15550915 TI - [Fungal prophylaxis following reduced-intensity stem cell transplantation (RIST)]. AB - Hematopoietic stem cell transplantation has been established as a curative treatment for advanced hematologic malignancies. Transplantation with a reduced intensity conditioning regimen has been developed. The minimal toxicity of reduced-intensity stem cell transplantation (RIST) has made transplantation available for patients of advanced age or with organ dysfunction. The response of malignant lymphoma and some solid tumors to RIST has been observed. RIST with unrelated donors and umbilical cord blood has been studied. Fungal infection is an important complication of RIST. Since the prognosis of fungal infection is poor, the management has been focused on its prophylaxis. Given recent progression in RIST management, the strategy of infectious prophylaxis has also changed. Equipment in the hospital is important for fungal infection; however, the median day of the development of fungal infection is day 100, when most patients are followed as outpatients. The focus of fungal management after RIST is oral antifungal agents rather than in-hospital equipment. Various antifungal agents have recently been developed and applied for clinical use. Many antifungals have been developed simultaneously for the first time. A major change in antifungal management will probably occur in the next several years. PMID- 15550916 TI - [Efficacy study of early presumptive therapy (EPT) for deep fungal infection]. AB - We retrospectively studied the efficacy of early presumptive therapy (EPT). SUBJECTS AND METHOD: Of the critically ill patients admitted from January 1998 to the end of December 2000 to Kyorin University Trauma Burn and Intensive Care Center, 77 cases were diagnosed with suspected deep fungal infection, and EPT was administered. The diagnosis of suspected deep fungal infection was made by definition. EPT (FLCZ 200 to 400 mg/day x 14 days) was started as soon as the diagnosis was made and continued for two weeks. Its efficacy was retrospectively studied by analyzing the clinical findings, changes in local organisms, and hematological tests. RESULTS: After treatment, 62% of the patients showed improvement in clinical signs of infection, elimination of locally detected fungus, and improvement in the serum diagnosis test. Post-EPT detection levels of the fungus had decreased to 21%. The mean pre-EPT body temperature was 38.7 degrees C +/- 0.6 degrees C, but the mean post-EPT temperature was 36.7 degrees C +/- 0.6 degrees C. The mean level of blood 1,3-beta-D-glucan was 35 plus minus 13 pg/ml at the time the diagnosis was made, but returned to normal levels after treatment had concluded. No patients died as a direct result of the fungal infection. CONCLUSION: This study of early presumptive therapy in critically ill patients in the emergency and intensive care medicine fields showed the therapy in these, and in high risk patients to be efficacious. PMID- 15550917 TI - [Guidelines for the management of deep mycosis in neutropenic patients]. AB - Invasive fungal infections (IFIs) are a major cause of morbidity and mortality in neutropenic patients with leukemia and those undergoing hematopoietic stem cell transplant (HSCT). Two major IFIs are systemic candidiasis (including candidemia, chronic disseminated candidiasis and pneumonia) and invasive pulmonary aspergillosis. Recently, the incidence of the latter has been increasing. Three levels of diagnosis are specified in the Japanese guidelines for the diagnosis and treatment of IFIs. Proven fungal infections are diagnosed by histological/microbiological evidence of fungi at the site of infection or positive blood culture (fungemia). Clinically documented fungal infections are diagnosed by typical radiological findings such as halo sign on chest CT plus positive serological/molecular evidence of fungi such as Aspergillus galactomannan, beta-glucan or fungal DNA. Possible fungal infections are diagnosed by typical radiological findings or positive serological/molecular evidence of fungi. For patients with high risk such as those undergoing HSCT, antifungal prophylaxis using oral antifungal agents is recommended. For possible fungal infections, empiric therapy with fluconazole (FLCZ) or amphotericin B (AMPH) is recommended. For patients with proven fungal infections or clinically documented fungal infections, targeted therapy is warranted. In case of candidemia, the best choice is FLCZ (400 mg/day) or AMPH (0.5-0.7 mg/kg/day), and for invasive pulmonary aspergillosis, a higher dose of AMPH (1.0-1.5 mg/kg/day) is indicated. Micafungin (MCFG), recently licensed in Japan, is an active agent for both Candida and Aspergillus. This drug seems useful for empiric and targeted therapy of IFIs. PMID- 15550918 TI - [Strategy for the treatment of fungal infections in critically ill surgical patients]. AB - To improve the outcome of invasive Candida infections, earlier empirical therapy before the establishment of the definitive diagnosis is considered to be necessary. However, appropriate use of empirical therapy for suspected candidiasis in febrile non-neutropenic surgical patients has not been defined. According to the guidelines from the Infectious Diseases Society of America, empirical therapy of suspected candidiasis in this setting should be limited to patients with Candida colonization of multiple sites, multiple other risk factors, and absence of any other causes of fever. A corrected colonization index which takes into account both the density and the degree of colonization of Candida spp. was shown to be the independent factors that predict subsequent candidal infection. It may also be appropriate to commence empirical therapy on the basis of a positive serodiagnostic test. Beta-D glucan is a cell-wall constituent of fungi, which is assumed to be a marker of fungal sepsis. However, it has been shown that beta-D-glucan can also be detected in patients without fungal infections, such as those on haemodialysis, and its positive predictive value is relatively low. The mono-utilization of beta-D-glucan for the assessment of fungal infection should therefore be avoided. The combined assessment of beta D-glucan and extent of colonization with Candida spp. is believed to have the advantage of lessening the likelihood of a false positive reaction of beta-D glucan. PMID- 15550919 TI - [Invasive mycosis in solid organ transplantation]. AB - Invasive mycosis in solid organ transplantation is mainly caused by Candida and Aspergillus, and its risk is higher in small bowel, liver, pancreas, and lung transplantation. Although limited analyses propose not a few risk factors for invasive mycosis in respective transplanted organs, the efficacy of prophylactic use of antifungal agents or preemptive treatments based on the information is not fully supported by prospective randomized controlled clinical data. The final guideline should be helpful for tailor-made evidence-based management based on the stratification of patients by pretransplant, surgical, immunosuppressive and organ specific characteristics. The process of repeated proposals and verification in a large number of patients is necessary. PMID- 15550920 TI - [Development of murine experimental model for candidiasis and its application]. AB - Candida albicans is a major cause of oral and esophageal infections in elder persons with poor oral hygiene and immuncompromised patients with hyposalivation, diabetes mellitus, prolonged use of antibiotics or immunosuppressive drugs. Oral thrush is a common form of oropharyngeal candidiasis whose clinical features consist of white patches appearing as discrete lesions on the buccal mucosa, throat, tongue, and gum linings that develop into confluent pseudomembranes resembling milk curds. We recently reported a simple murine model of thrush-type oral candidiasis that mimics the natural infectionin humans and is useful for both symptomatological and mycological evaluation of the responsiveness to antifungal treatments. By using this oral candidiasis model, protective activities of oral administration of several types of herbal preparations such as teatree oil, clove preparation and bovine lactoferrin were clarified. The mechanisms of protective actions of lactoferrin against oral candidiasis were particularly elucidated to include augmentation of T-cell activities of lesional lymphoid tissues. More recent studies suggested that saliva from healthy persons also shows a protective action for this murine oral candidiasis model. PMID- 15550921 TI - [Clinical and fundamental investigations on recurrent glossodynia]. AB - A type of oral lesion, so-called glossodynia has been on the increase recently. Glossodynia is a kind of psychosomatic disease in which the patient experiences chronic pain on the surface of the tongue. It has never been diagnosed as coming from organic or functional pain. Although glossodynia can be cured by antianxiety drugs, antidepressants, or autogenic training and so on, usually. These are not a desirable solution. We initially tried to administer the antifungal drug, ITCZ, to 65 glossodynia patients. Sixty-four of them were cured of tongue pain after 1 3 weeks. The effective rate of recovery was 98.5%. Only two patients experienced recurrence of pain after 15 and 17 weeks, respectively, and Candida albicans was isolated from the surface of their tongues. The nature of the recurrent strains was investigated by MICs against 4 antifungal drugs, ITCZ, MCZ, AMPH-B, and NYS, as well as by the API 20C Auxanogram biochemically, and a molecular epidemiologic study by PFGE. Each case of Candida albicans was almost the same before and after the administration of ITCZ. Above all, it is important to carefully inspect the candidiasis of the tongue and to initially administer antifungal drugs when the diagnosis is glossodynia. PMID- 15550922 TI - [A case of allergic fungal sinusitis caused by Bipolaris spicifera]. AB - We describe a case of allergic fungal sinusitis (AFS) caused by Bipolaris spicifera, the first case reported in Japan. A 70-year-old man was admitted to our hospital because of diplopia following bilateral nasal obstruction and discharge. Radiological studies using computed tomographic scan showed a large soft tissue mass occupying the right frontal, bilateral ethmoid and sphenoid sinuses. He underwent drainage surgery and histopathological examination of the contents of the paranasal sinuses revealed scattered fungal hyphae within "allergic mucin". By cytological examination, these hyphae showed septation at irregular intervals, and were swollen to various sizes. Microbiological studies identified the fungus as Bipolaris spicifera. The presence of allergic mucin and scattered fungal hyphae were very important findings in making a diagnosis of AFS histopathologically, so squash cytology of the contents of the paranasal sinuses was quite useful to observe fungal elements and identify the strain in this case. PMID- 15550923 TI - [Experimental studies on the penetration of dermatophytes into the human stratum corneum and the effects of washing]. AB - After several types of dermatophytes were applied to the stratum corneum obtained from a healthy human heel, we evaluated the penetration speed of fungal elements into the stratum corneum and the effect of washing its surface. We designed the following two environments, assuming tinea pedis after applying fungal elements to the surface of the stratum corneum. The samples were incubated under conditions simulating the daily life of those who wear socks in a house: (1) 90% humidity for 8 hours, and 100% humidity for 16 hours, and those who have bare feet in a house: (2) 80% humidity for 8 hours and 100% humidity for 16 hours. We took the samples out every 24 hours and made observations by PAS stain and scanning electron microscope before and after washing them. Although fungal elements were not removable in (1), even if washed one day later, they could be removable one and two days later in (2). We suggest that fungal elements were easily removable even if dermatophytes had begun to penetrate the surface of the stratum corneum, because the soles retained a low humidity when the shoes were removed, and the soles and interdigital regions were washed every day. Moreover, in an experiment assuming tinea corporis, fungal elements were applied to the cutting side of the stratum corneum, incubated at 80% humidity, and observed after PAS stain. The penetration of Trichophyton tonsurans to the cutting side of the stratum corneum began in 0.5 days, which was sooner than other dermatophytes. We believe this is one factor of the latest expansion of T. tonsurans infection in Japan. PMID- 15550924 TI - Efficient engraftment of in utero transplanted mice with retrovirally transduced hematopoietic stem cells. AB - Using an experimental mouse model, we have investigated the kinetics of hematopoietic reconstitution of recipients transplanted during fetal development with fresh and transduced hematopoietic stem cells (HSCs). Total bone marrow (BM) and purified Lin(-)Sca-1(+) cells, either fresh or transduced ex vivo with enhanced green fluorescent protein (EGFP)-encoding retroviral vectors, were in utero transplanted (IUT) into fetal mice. Data obtained 2 months after transplantation showed a similar proportion of engrafted animals, regardless of the fact that samples were purified or not on HSCs, and subjected or not to ex vivo transduction with retroviral vectors. The transplantation of grafts enriched in HSCs, either fresh or transduced, always improved the levels of donor chimerism of IUT mice in comparison with results obtained in mice transplanted with unpurified BM grafts (6.8 and 7.3% versus 1.15% median values, respectively). Significantly, engrafted recipients that were transplanted with the transduced graft always contained transduced EGFP(+) cells in peripheral blood (around 5% of donor cells were EGFP(+) at 2 months post-transplantation). This proportion was essentially maintained at longer times post-transplantation, as well as in secondary recipients transplanted with the BM of IUT mice. Our study describes for the first time a significant and stable engraftment of unconditioned mice subjected to IUT with HSCs transduced with retroviral vectors. PMID- 15550925 TI - Comparative studies on cellular gene regulation by HIV-1 based vectors: implications for quality control of vector production. AB - Among the recent gene therapy protocols, vectors that can stably express transgenes, for example, HIV-1-based vectors, are particularly desirable. There have been no direct reports on insertional mutagenesis by lentiviral vectors; however, the severe pathogenic nature of their parental virus (HIV-1) is still a major safety concern surrounding these vectors and prevents the progress with their clinical application. We reason that by investigating the host response we shall be able to assess the safety and potential effects of the vectors on targeted cells and understand the interaction between vectors and the host. For this, two major sets of experiments were conducted. Initially, we used cDNA microarray methodology to examine cellular gene profile in human primary umbilical cord endothelial cells (HUVECs) after HIV-1-based VSV-G/GFP vector transduction and observed a modest effect of HIV-1-based vectors on HUVECs. The represented functional categories include transcription and translation factors, tumour antigens, complement factors and signal transduction factors. Some of the differentially expressed genes, for example, Clusterin, CD151, Ku antigen and eIF4gamma, could have oncogenic potential. In the second approach, we systematically compared five different viral vectors, that is, HIV-1-based VSV G/Empty, VSV-G/GFP, VSV-G/puro, Amph/GFP and MLV-based Amph/Laz, for the effects of individual viral components on cellular gene regulation. Our comparative results demonstrated a regulatory function of Gag/Pol proteins on cellular gene expression. The significance of our findings in relation to the safety of HIV-1 vectors and the importance of quality control of vector production will be presented and discussed. PMID- 15550926 TI - Complement regulatory proteins are incorporated into lentiviral vectors and protect particles against complement inactivation. AB - Lentiviral vectors pseudotyped with G glycoprotein from vesicular stomatitis virus (VSV-G) and baculovirus gp64 are inactivated by human complement. The extent of vector inactivation in serum from individual donors was examined and results showed wide donor-dependent variation in complement sensitivity for VSV-G pseudotyped lentivectors. Amphotropic envelope (Ampho)-pseudotyped vectors were generally resistant to serum from all donors, while gp64-pseudotyped vectors were inactivated but showed less donor-to-donor variation than VSV-G. In animal sera, the vectors were mostly resistant to inactivation by rodent complement, whereas canine complement caused a moderate reduction in titer. In a novel advance for the lentiviral vector system, human complement-resistant-pseudotyped lentivector particles were produced through incorporation of complement regulatory proteins (CRPs). Decay accelerating factor (DAF)/CD55 provided the most effective protection using this method, while membrane cofactor protein (MCP)/CD46 showed donor-dependent protection and CD59 provided little or no protection against complement inactivation. Unlike previous approaches using CRPs to produce complement-resistant viral vectors, CRP-containing lentivectors particles were generated for this study without engineering the CRP molecules. Thus, through overexpression of native DAF/CD55 in the viral producer cell, an easy method was developed for generation of lentiviral vectors that are almost completely resistant to inactivation by human complement. Production of complement-resistant lentiviral particles is a critical step toward use of these vectors for in vivo gene therapy applications. PMID- 15550927 TI - Adeno-associated virus-mediated delivery of a mutant endostatin suppresses ovarian carcinoma growth in mice. AB - Earlier studies have shown that a point mutation in human endostatin at position 125 (human endostatin wherein proline 125 was substituted with alanine, P125A endostatin) improves endothelial cell binding and antiangiogenic activity. In the present study, we investigated the effect of recombinant adeno-associated virus (rAAV)-mediated gene delivery of P125A-endostatin (rAAV-P125Aendo) in a mouse model of ovarian carcinoma. Intramuscular (i.m.) injection of rAAV-P125Aendo resulted in a dose-dependent increase in serum endostatin levels. Consequently, vascular endothelial growth factor- and basic fibroblast growth factor-mediated angiogenesis was significantly inhibited in mice injected with rAAV-P125Aendo as compared to control mice injected with rAAV-LacZ. Furthermore, gene therapy using rAAV-P125Aendo construct showed sustained secretion of P125A-endostatin for up to 9 weeks after a single i.m. administration. Recombinant AAV-P125Aendo injection significantly inhibited the growth of human ovarian cancer cells in athymic nude mice. Immunofluorescence studies of residual tumors surgically removed from the rAAV-P125Aendo-treated animals showed decreased number of vessel ends and vessel length, indicating inhibition of angiogenesis. These studies suggest that recombinant AAV-mediated antiangiogenic gene therapy methods can be used to inhibit ovarian cancer growth. PMID- 15550928 TI - Optimization of equine infectious anemia derived vectors for hematopoietic cell lineage gene transfer. AB - Gene transfer into hematopoietic cells may allow correction of a variety of hematopoietic and metabolic disorders. Optimized HIV-1 based lentiviral vectors have been developed for improved gene transfer and transgene expression into hematopoietic cells. However, the use of HIV-1 based vectors for human gene therapy may be limited due to ethical and biosafety issues. We report that vectors based on the non-primate equine infectious anemia virus (EIAV) transduce a variety of human hematopoietic cell lines and primary blood cells. To investigate optimization of gene expression in hematopoietic cells, we compared a variety of post-transcriptional elements and promoters in the context of EIAV vectors. We observed cell specific increase in the number of transgene expressing cells with the different post-transcriptional elements, whereas the use of elongation factor alpha 1 (EFalpha1) promoter resulted in significant increases in both the number of transgene expressing cells and the level of transgene protein in all cell types tested. We then demonstrate increased transduction of hematopoietic cells using a second-generation EIAV vector containing a self inactivating EIAV LTR and the EIAV central polypurine tract (cppt). These data suggest that optimized EIAV vectors may be a suitable alternative to HIV-1 vectors for use in hematopoietic gene therapy. PMID- 15550929 TI - The magic of synaptogenesis. Meeting on spinogenesis and synaptic plasticity. PMID- 15550930 TI - Replication-independent chromatin loading of Dnmt1 during G2 and M phases. AB - The major DNA methyltransferase, Dnmt1, associates with DNA replication sites in S phase maintaining the methylation pattern in the newly synthesized strand. In view of the slow kinetics of Dnmt1 in vitro versus the fast progression of the replication fork, we have tested whether Dnmt1 associates with chromatin beyond S phase. Using time-lapse microscopy of mammalian cells expressing green fluorescent-protein-tagged Dnmt1 and DsRed-tagged DNA Ligase I as a cell cycle progression marker, we have found that Dnmt1 associates with chromatin during G2 and M. This association is mediated by a specific targeting sequence, shows strong preference for constitutive but not facultative heterochromatin and is independent of heterochromatin-specific histone H3 Lys 9 trimethylation, SUV39H and HP1. Moreover, photobleaching analyses showed that Dnmt1 is continuously loaded onto chromatin throughout G2 and M, indicating a replication-independent role of Dnmt1 that could represent a novel and separate pathway to maintain DNA methylation. PMID- 15550931 TI - Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator. AB - Epac1 is a guanine nucleotide exchange factor for Rap1 that is activated by direct binding of cAMP. In vitro studies suggest that cAMP relieves the interaction between the regulatory and catalytic domains of Epac. Here, we monitor Epac1 activation in vivo by using a CFP-Epac-YFP fusion construct. When expressed in mammalian cells, CFP-Epac-YFP shows significant fluorescence resonance energy transfer (FRET). FRET rapidly decreases in response to the cAMP raising agents, whereas it fully recovers after addition of cAMP-lowering agonists. Thus, by undergoing a cAMP-induced conformational change, CFP-Epac-YFP serves as a highly sensitive cAMP indicator in vivo. When compared with a protein kinase A (PKA)-based sensor, Epac-based cAMP probes show an extended dynamic range and a better signal-to-noise ratio; furthermore, as a single polypeptide, CFP-Epac-YFP does not suffer from the technical problems encountered with multisubunit PKA-based sensors. These properties make Epac-based FRET probes the preferred indicators for monitoring cAMP levels in vivo. PMID- 15550932 TI - The effect of etanercept on Graves' ophthalmopathy: a pilot study. AB - PURPOSE: To assess the effect of etanercept, an antitumour necrosis factor (anti TNF) drug, on clinical signs in Graves' ophthalmopathy (GO). METHODS: A total of 10 consecutive patients (seven female, three male) with recent-onset, active, mildly-to-moderately severe GO were treated with subcutaneous injections of 25 mg etanercept (Enbrel) twice weekly during 12 weeks. The average age was 50 years (range: 39-59 years). The average duration of GO was 4 months (range: 2-6 months). All patients were at least 2 months euthyroid with medication. The mean pretreatment clinical activity score (CAS) was 4 (range: 3-6). RESULTS: The mean CAS prior to treatment was 4, at 6 weeks 2.6, and at 12 weeks 1.6. The mean ophthalmopathy index (OI) prior to treatment was 5.8, at 6 weeks 4.8, and at 12 weeks 4.4. Thus, after 12 weeks of treatment the mean CAS had decreased by 2.4+/ 1.3 (60%), and the mean OI had decreased by 1.4+/-1.2 (24%). The main decrease of the CAS and OI was noted at 6 weeks of treatment. The difference was particularly evident on soft-tissue changes, including periocular chemosis and redness. The mean exophthalmometry values did not change. In all, 60% percent of patients reported moderate to marked improvement. In three patients, the GO flared up again after cessation of treatment. There were no serious adverse events or side effects during a mean follow-up time of 18 months. CONCLUSIONS: This study strongly suggests that etanercept may suppress the clinical signs in GO, but randomized controlled trials are needed to further evaluate the effect of anti TNF treatment and to compare its side-effects with those of corticosteroids. PMID- 15550933 TI - Anterior ischaemic optic neuropathy secondary to Henoch-Schonlein Purpura. PMID- 15550934 TI - Morphology of corneal nerves and corneal sensation in dry eye: a preliminary study. AB - PURPOSE: To evaluate the corneal sensitivity and nerve morphology in dry eyes. METHODS: A total of 32 eyes of 16 patients (10 Sjogren's syndrome and six non Sjogren's syndrome) and 19 eyes of 10 age-matched controls were studied. Sensitivity of the central cornea was measured by the Cochet-Bonnet aesthesiometer. The morphology of corneal nerves was studied by in vivo confocal microscopy (ConfoScan 2.0, Fortune Technologies Srl, Vigonza (PD), Italy). Sub basal epithelial nerve plexus, subepithelial nerve plexus, and stromal nerves were localized and evaluated for the number of nerves, thickness, reflectivity, and tortuosity for each frame. RESULTS: The mean corneal sensitivity of dry eye patients (5.6 mm/grs/S) was found significantly lower than that of the control (5.0 mm/grs/S) group (P<0.01). Although the subepithelial nerves were thicker in dry eyes (mean: 6.38+/-1.18 microm) than control eyes (mean: 5.72+/-1.27 microm), this difference was not significant (P>0.05). CONCLUSION: Decreased corneal sensitivity is not associated with morphological changes of corneal nerves in dry eyes. PMID- 15550935 TI - Different apoptotic mechanisms are involved in the antiproliferative effects of 7beta-hydroxysitosterol and 7beta-hydroxycholesterol in human colon cancer cells. AB - Plant sterols are found in fruits and vegetables. Their cholesterol-lowering effect is well documented. Our study aimed at comparing antiproliferative effects of 7beta-hydroxysitosterol (7beta-OHsito) versus 7beta-hydroxycholesterol (7beta OHchol) on the human colon cancer cell line Caco-2. When cells were exposed for 32 h to 60 microM 7beta-OHsito or to 30 microM 7beta-OHchol, both compounds caused 50% growth inhibition. Cells treated with 7beta-OHsito showed enhanced caspase-9 and -3 activities followed by DNA fragmentation. In contrast, 7beta OHchol did not activate caspase-3 and activation of caspase-9 and DNA fragmentation were delayed. The treatment of cells with the caspase inhibitor Z VAD.fmk retarded the 7beta-OHsito-induced apoptotic process but not that triggered by 7beta-OHchol. Our data suggest that the two compounds in spite of their structural similarities target different cellular pathways, which lead to cell death. PMID- 15550936 TI - The post-translational phosphorylation and acetylation modification profile is not the determining factor in targeting endogenous stress-induced p53 to mitochondria. PMID- 15550937 TI - Mechanisms of resistance to TRAIL-induced apoptosis in cancer. AB - The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is regarded as a potential anticancer agent. However, considerable numbers of cancer cells, especially some highly malignant tumors, are resistant to apoptosis induction by TRAIL, and some cancer cells that were originally sensitive to TRAIL-induced apoptosis can become resistant after repeated exposure (acquired resistance). Understanding the mechanisms underlying such resistance and developing strategies to overcome it are important for the successful use of TRAIL for cancer therapy. Resistance to TRAIL can occur at different points in the signaling pathways of TRAIL-induced apoptosis. Dysfunctions of the death receptors DR4 and DR5 due to mutations can lead to resistance. The adaptor protein Fas-associated death domain (FADD) and caspase-8 are essential for assembly of the death-inducing signaling complex, and defects in either of these molecules can lead to TRAIL resistance. Overexpression of cellular FADD-like interleukin-1beta-converting enzyme inhibitory protein (cFLIP) correlates with TRAIL resistance in several types of cancer. Overexpression of Bcl-2 or Bcl-X(L), loss of Bax or Bak function, high expression of inhibitor of apoptosis proteins, and reduced release of second mitochondria-derived activator of caspases (Smac/Diablo) from the mitochondria to the cytosol have all been reported to result in TRAIL resistance in mitochondria dependent type II cancer cells. Finally, activation of different subunits of mitogen-activated protein kinases or nuclear factor-kappa B can lead to development of either TRAIL resistance or apoptosis in certain types of cancer cells. PMID- 15550938 TI - Short interfering RNAs as a tool for cancer gene therapy. AB - There are mainly two types of short RNAs that target complementary messengers in animals: small interfering RNAs and micro-RNAs. Both are produced by the cleavage of double-stranded RNA precursors by Dicer, a member of the Rnase III family of double-stranded specific endonucleases, and both guide the RNA-induced silencing complex to cleave specifically RNAs sharing sequence identity with them. In designing a particular RNA interference (RNAi), it is important to identify the sense/antisense combination that provides the most potent suppression of the target mRNA, and several rules have been established to give >90% gene expression inhibition. RNAi technology can be directed against cancer using a variety of strategies. These include the inhibition of overexpressed oncogenes, blocking cell division by interfering with cyclin E and related genes or promoting apoptosis by suppressing antiapoptotic genes. RNAi against multidrug resistance genes or chemoresistance targets may also provide useful cancer treatments. Studies investigating these approaches in preclinical models are also reviewed. PMID- 15550939 TI - The evolution of chronic infection strategies in the alpha-proteobacteria. AB - Many of the alpha-proteobacteria establish long-term, often chronic, interactions with higher eukaryotes. These interactions range from pericellular colonization through facultative intracellular multiplication to obligate intracellular lifestyles. A common feature in this wide range of interactions is modulation of host-cell proliferation, which sometimes leads to the formation of tumour-like structures in which the bacteria can grow. Comparative genome analyses reveal genome reduction by gene loss in the intracellular alpha-proteobacterial lineages, and genome expansion by gene duplication and horizontal gene transfer in the free-living species. In this review, we discuss alpha-proteobacterial genome evolution and highlight strategies and mechanisms used by these bacteria to infect and multiply in eukaryotic cells. PMID- 15550940 TI - Iron and microbial infection. AB - The use of iron as a cofactor in basic metabolic pathways is essential to both pathogenic microorganisms and their hosts. It is also a pivotal component of the innate immune response through its role in the generation of toxic oxygen and nitrogen intermediates. During evolution, the shared requirement of micro- and macroorganisms for this important nutrient has shaped the pathogen-host relationship. Here, we discuss how pathogens compete with the host for iron, and also how the host uses iron to counteract this threat. PMID- 15550941 TI - Bacterial redox sensors. AB - Redox reactions pervade living cells. They are central to both anabolic and catabolic metabolism. The ability to maintain redox balance is therefore vital to all organisms. Various regulatory sensors continually monitor the redox state of the internal and external environments and control the processes that work to maintain redox homeostasis. In response to redox imbalance, new metabolic pathways are initiated, the repair or bypassing of damaged cellular components is coordinated and systems that protect the cell from further damage are induced. Advances in biochemical analyses are revealing a range of elegant solutions that have evolved to allow bacteria to sense different redox signals. PMID- 15550942 TI - Tularaemia: bioterrorism defence renews interest in Francisella tularensis. AB - Francisella tularensis is a highly infectious aerosolizable intracellular pathogen that is capable of causing a debilitating or fatal disease with doses as low as 25 colony-forming units. There is no licensed vaccine available. Since the 1950s there has been concern that F. tularensis could be used as a biological threat agent, and it has received renewed attention recently owing to concerns about bioterrorism. The International Conference on Tularaemia in 2003 attracted more than 200 delegates, twice the number of participants as previous meetings. This is a reflection of the increased funding of research on this pathogen, particularly in the United States. PMID- 15550943 TI - Epidemiological interpretation of antibiotic resistance studies - what are we missing? AB - Antimicrobial resistance is an emerging public-health threat. Studies of the relationship between antibiotic use and resistance, as well as surveillance programmes, examine changes in the proportion of isolates that are resistant. Although proportions are helpful to the clinician prescribing empirical therapy, proportion-based analyses can be misleading to the public-health professional as they can yield biased estimates. Proportions do not adequately reflect the burden of resistance, a measure often of interest in public health. A more appropriate measure of this burden is the rate of isolation of resistant organisms, that is, the absolute number of resistant isolates in a population over time. PMID- 15550944 TI - The life and times of ivermectin - a success story. AB - Since its introduction more than 20 years ago, ivermectin has proved to be one of the most successful therapeutic drugs in veterinary medicine, as well as the basis of one of the most successful public-health programmes of the past century. The drug arose from a unique international collaboration between the public and private sectors. The development process also incorporated the world's first and largest drug-donation programme and involved a unique association between governments, non-governmental organizations and industry. The drug is now being used, free of charge, in two global disease-elimination programmes that are benefiting millions of the world's poorest people. PMID- 15550945 TI - Antibiotic resistance: a view from the prescriber. AB - Antibiotic resistance is increasingly affecting the management of infectious diseases. The prescribing clinician is not only important to the development of the problem but also central to its solution. This article addresses the current weaknesses in the information systems and the evidence base that support prescribing. Remedies necessary for improvements in prudent prescribing include better guidance in managing specific diseases where resistance is of prognostic significance and also increasing diagnostic precision. PMID- 15550946 TI - Towards a structural view of gating in potassium channels. AB - Voltage-activated cation channels have pores that are selective for K(+), Na(+) or Ca(2+). Neurons use these channels to generate and propagate action potentials, release neurotransmitters at synaptic terminals and integrate incoming signals in dendrites. Recent X-ray and electron microscopy studies of an archaebacterial voltage-activated K(+) (Kv) channel have provided the first atomic resolution images of the voltage-sensing domains in Kv channels. Although these structures are consistent with previous biophysical analyses of eukaryotic channels, they also contain surprises, which have provoked new ideas about the structure and movements of these proteins during gating. This review summarizes our current understanding of these intriguing membrane proteins and highlights the open questions. PMID- 15550947 TI - Stress, cognitive impairment and cell adhesion molecules. AB - Stress has profound effects on brain structure and function, but the underlying mechanisms are still poorly understood. Recent studies imply that neuronal cell adhesion molecules of the immunoglobulin superfamily--NCAM and L1--are important mediators of the effects of stress on the brain. Chronic stress regimes that lead to hippocampal atrophy and spatial-learning impairment in rodents simultaneously induce a pattern of changes in cell adhesion molecule expression that fits with a role for these molecules in stress-induced neuronal damage and neuroprotective mechanisms. These findings highlight cell adhesion molecules as potential therapeutic targets to treat stress-related cognitive disturbances. PMID- 15550948 TI - Biochemical mechanisms for translational regulation in synaptic plasticity. AB - Changes in gene expression are required for long-lasting synaptic plasticity and long-term memory in both invertebrates and vertebrates. Regulation of local protein synthesis allows synapses to control synaptic strength independently of messenger RNA synthesis in the cell body. Recent reports indicate that several biochemical signalling cascades couple neurotransmitter and neurotrophin receptors to translational regulatory factors in protein synthesis-dependent forms of synaptic plasticity and memory. In this review, we highlight these translational regulatory mechanisms and the signalling pathways that govern the expression of synaptic plasticity in response to specific types of neuronal stimulation. PMID- 15550949 TI - Insights into neural mechanisms and evolution of behaviour from electric fish. AB - Both behaviour and its neural control can be studied at two levels. At the proximate level, we aim to identify the neural circuits that control behaviour and to understand how information is represented and processed in these circuits. Ultimately, however, we are faced with questions of why particular neural solutions have arisen, and what factors govern the ways in which neural circuits are modified during the evolution of new behaviours. Only by integrating these levels of analysis can we fully understand the neural control of behaviour. Recent studies of electrosensory systems show how this synthesis can benefit from the use of tractable systems and comparative studies. PMID- 15550950 TI - Receptor trafficking and synaptic plasticity. AB - Long-term potentiation and long-term depression are processes that have been widely studied to understand the molecular basis of information storage in the brain. Glutamate receptors are required for the induction and expression of these forms of plasticity, and GABA (gamma-aminobutyric acid) receptors are involved in their modulation. Recent insights into how these receptors are rapidly moved into and out of synaptic membranes has profound implications for our understanding of the mechanisms of long-term potentiation and long-term depression. PMID- 15550951 TI - Drug addiction: the neurobiology of behaviour gone awry. AB - Drug addiction manifests as a compulsive drive to take a drug despite serious adverse consequences. This aberrant behaviour has traditionally been viewed as bad "choices" that are made voluntarily by the addict. However, recent studies have shown that repeated drug use leads to long-lasting changes in the brain that undermine voluntary control. This, combined with new knowledge of how environmental, genetic and developmental factors contribute to addiction, should bring about changes in our approach to the prevention and treatment of addiction. PMID- 15550952 TI - [What we don't talk about]. PMID- 15550953 TI - [Death in nursing homes]. PMID- 15550955 TI - [Increased health inequality?]. PMID- 15550954 TI - [Sleep disorders should be investigated for treatment]. PMID- 15550956 TI - [Employment trends--the impact of chronic disease in childhood and low educational level]. AB - BACKGROUND: Our aim was to analyse long-term employment trends among disadvantaged groups on the assumption that they may be more exposed to unemployment during recessions or be at increasingly higher risk of unemployment because of stricter requirements in the workplace. MATERIAL AND METHODS: We established a cohort through links between data on individuals in national registers for all 321,975 men born 1967 through 1976. Disadvantaged groups under study were those receiving supplementary benefits in childhood because of chronic disease, and those with few educational attainments. Using Poisson regression we estimated associations with unemployment or disability over the course of the 1991-2001 period. RESULTS: Disadvantaged groups had higher risk of unemployment or disability. Associations between chronic disease in childhood and subsequent unemployment followed the business cycle; they were highest during the 1993 recession and lowest during the boom years around 1999. Over the long term, the gap in unemployment widened between those with low educational attainment and the better-educated. We found similar but less clear trends for disability. INTERPRETATION: The observed negative effect of chronic childhood disease on employment status paralleled the business cycle: it was weakest in boom years and strongest in recession years. Low educational attainment was associated with a gradual decline in the level of employment. PMID- 15550957 TI - [Could persons on sick leave learn to think differently about themselves and increase their participation in the workplace?]. AB - BACKGROUND: The aim of this study was to evaluate an empowerment programme for improved quality of life and ability to work. At two places of work, a total of 112 employees with frequent sick-leaves over the last six months were invited to participate; 19 women attended. After the intervention the participants reported better coping and quality of life, and they worked more hours per week than previously. In their opinion, the improvement was due to the intervention. This paper relates their experiences and gives an account of their work participation one year later. MATERIAL AND METHODS: Questionnaires by the start of and soon after the intervention, telephone interviews and focus group interviews. RESULTS: Only minor differences were found after one year compared to just after the intervention. Nearly all the participants expressed more self-esteem and worked more hours per week. They particularly emphasised their new work consciousness and the benefit from the counselling received. INTERPRETATION: The results in this small group indicate that after one year the participants had kept most of the self-reconstruction of life that seemed to be achieved during the intervention. PMID- 15550958 TI - [Clinical trials in Norway--completion and reporting are not satisfactory]. AB - OBJECTIVE: To investigate how drug trials are carried out and reported in Norway and to what extent they are published. MATERIAL AND METHODS: All drug trials notified in 1996 were included in the study. Data were obtained from the standard notification form, correspondence with investigators, end-of-study reports, and a questionnaire designed for this study. RESULTS: A total of 208 drug trials were notified. Most trials were initiated by the pharmaceutical industry (85%) and international multicenter studies constituted a major part (73%). Mandatory end of-study reports were submitted to the health authorities on 48 (23%) of the trials. Out of a total of 159 trials for which we have data, 39 (25%) were interrupted or not started. Out of a total of 143 trials for which we have data on publishing, 77 (54%) were not published. Trials with a positive conclusion (54%) were more likely to be published than those with a negative conclusion (38%). INTERPRETATION: The reporting of drug trials is not satisfactory. Because of low reporting frequency, health authorities do not obtain a comprehensive overview. The pharmaceutical industry initiates the majority of the trials and clinical researchers in Norway increasingly participate in international multicentre trials. Many trials are not carried out as planned; less than half are published. PMID- 15550959 TI - [Dissemination of evidence-based information about PSA test and prostatic cancer to physicians]. AB - BACKGROUND: This paper describes the evaluation of an initiative to disseminate to general practitioners, urologists and patients in Norway evidence from systematic reviews on the clinical effectiveness of prostate cancer screening. Nine systematic reviews concluded that there is no evidence that early detection and treatment of prostate cancer reduces mortality or morbidity or improve patients' quality of life. MATERIALS AND METHODS: Evidence-based information about prostrate-specific antigen and prostate cancer, including patient information leaflets, was disseminated to 4100 general practitioners and specialists in urology in Norway in March 2001. One year later they were mailed a questionnaire aimed at evaluating how they had perceived and implemented this information. RESULTS: The results of the survey (response rate 41%) showed a marked contrast between general practitioners and urologists. 91% of general practitioners versus 61% of urologists agreed to the recommendation that prostrate-specific antigen tests should not be taken by healthy men. 87% of general practitioners but only 33% of urologists followed up the recommendations in their practice. INTERPRETATION: Dissemination of evidence-based information about prostrate-specific antigen and prostate cancer was appreciated by general practitioners, had the power to convince and contributed to enhanced knowledge of a controversial issue, thereby increasing their confidence in their clinical decision making. PMID- 15550960 TI - [Health effects of outdoor air pollution]. AB - BACKGROUND: This paper addresses the contribution from epidemiological research to our current understanding of the health effects of ambient air pollution. MATERIAL AND METHODS: Well-known examples from the international literature are presented in order to describe the most commonly used research designs and show how such studies have contributed to our knowledge. A review of Norwegian studies is presented. RESULTS: A large number of studies have shown that quite low levels of air pollution are associated with negative health effects. Most studies have assessed short-term effects and both respiratory and cardiovascular effects have been shown to be associated with exposure to air pollutants. INTERPRETATION: The general view is that particulate matter causes the adverse health effect of air pollution, even though epidemiological studies alone have obvious limitations when it comes to conclusions about causation. Few Norwegian air pollution studies have so far been carried out; estimates of the health effects of air pollution in Norway must mainly rely on studies carried out in places with different climate and air pollution composition. PMID- 15550961 TI - [Self-management in anticoagulation--a meta-analysis]. AB - BACKGROUND: The aim of anticoagulation treatment is to reach an optimal international normalised ratio (INR) level in which the risk of thrombosis as well as bleeding is minimal. Recently developed methods that allow self-managed control of anticoagulation make more frequent testing possible and may thus represent a better alternative than conventional control. MATERIAL AND METHODS: In this meta-analysis, randomized, controlled trials comparing self-management with conventional control with regard to time in therapeutic range and number of bleedings and thromboses were investigated. Eight clinical trials fulfilled the criteria for inclusion, representing about 870 patient-years and 1300 patients. RESULTS: The difference in time in therapeutic range was on average 10% in favour of self-management. The rate of major and fatal bleedings was 3.5% per patient years with conventional control, 2.4% per patient-years with self-management. The rate of thrombosis was 3.9% per patient-years with conventional control as against 2.7% with self-management. OddsratioMH for major clinical events was 0.62 (95% CI 0.429, 0.895); p < 0.01. INTERPRETATION: This meta-analysis shows that self-managed control of anticoagulation improves safety and efficacy and is probably a better alternative than conventional control for selected groups of patients. PMID- 15550962 TI - [Parents' education and infant mortality 1967-1998]. AB - BACKGROUND: We have examined the association between socioeconomic status and risk of infant death in Norway between 1967 and 1998. METHODS: Information from the Medical Birth Registry of Norway on all live births and infant deaths was linked to information from Statistics Norway on parents' education. There were 1,777,364 eligible live births and 15,517 infant deaths. Differences between educational-attainment groups were estimated as risk difference, relative risk, population-attributable fraction, and index of inequality ratio. RESULTS: The risk of infant death decreased in all educational-attainment groups and the level of education increased over time. For neonatal (0-27 days of life) death the risk difference between infants whose mothers had high or low education was reduced from 3.5/1000 in the 1970s to 0.9/1000 in the 1990s. The inequality ratio declined from 1.72 to 1.32 and the population-attributable fraction from 22.3 to 8.4. For risk of postneonatal (28-364 days of life) death, the difference between infants whose mothers were in high or low education brackets increased from 0.7/1000 in the 1970s to 2.0/1000 in the 1990s. The inequality ratio went up from 1.31 to 4.00 and the population-attributable fraction from 9.7 to 39.5. INTERPRETATION: Since the late 1960s, infant mortality has decreased and parental educational levels have risen. There is a higher degree of social equality with regard to risk of neonatal death, while the opposite holds for postneonatal death. PMID- 15550963 TI - [Drug interactions in an intensive care unit]. AB - BACKGROUND: A new computerised interaction database in Norwegian is available at www.druid.uio.no and in some computer physician order entry systems. The scope of this paper was to assess drug use and examine the potential usefulness of DRUID in an intensive care unit. MATERIAL AND METHODS: Drugs given patients during their first 24 hours in our mixed surgical intensive care unit over one year were entered into DRUID. RESULTS: 191 patients received 1413 drugs (median 7). According to DRUID, 110 patients (58%) had 274 potential drug interactions (median 2) of 72 classes. No interaction was of grade A (should not be combined) or C (absorption of oral drugs), 153 were of grade B (take precautions), and 121 were of grade D (academic interest). Of grade B interactions, 51% were in two and 49% in 39 classes of drug interactions. Of grade D interactions, 60% in 4 and 40% in 27 classes. INTERPRETATION: Surprisingly few serious drug interactions were found. However, in every fourth patient precautions should be taken (e.g. dose adjustments). Electronic prescribing systems might give fewer adverse effects. PMID- 15550964 TI - [Anaphylactic reactions after intake of products with wheat]. AB - BACKGROUND: Food allergy is common in small children. Sensitisation to wheat is frequent, reactions to ingested wheat more seldom and mostly associated with atopic eczema and exercise-induced anaphylaxis. MATERIAL AND METHODS: Eleven children with severe reactions after ingestion of small amounts of wheat were referred to us for further diagnostic procedures. Skin prick tests, total and specific IgE as well as oral provocation test were performed. RESULTS: All but two children had strongly positive skin prick tests results to wheat. The same two children had specific IgE to Immunocap f4 (a-amylase) < 10 kUa/L while the nine other children had specific IgE to f4 > 29 kUa/L. All children had general reactions when provoked orally with small amounts of wheat with the need for administration of adrenalin and in most cases oral steroids. INTERPRETATION: It is important to identify children at risk for general reactions to wheat, as cereals are staple foods in our community. Identification of the proteins involved in serious reactions is essential. PMID- 15550965 TI - [Reasons for medical disqualification of Norwegian commercial pilots]. AB - BACKGROUND: The aim of this study is to present the reasons for medical disqualification (grounding) of Norwegian commercial pilots over a twenty-year period. We have also examined the issues of whether a change has taken place in the disease spectrum and to which extent new regulations have influenced aeromedical decision making. MATERIAL AND METHODS: The material is collected from the aeromedical section's archive. Files on all grounded pilots have been reviewed and classified by age group and diagnosis and grounding rates have been calculated. RESULTS: From the study population comprising 48,229 pilot-years, 275 pilots were permanently grounded, i.e. an average grounding rate of 5.7 per 1000 pilot-years. INTERPRETATION: Over this twenty-year period, the grounding rate based on cardiovascular disease fell; this may be explained by better treatment or more lenient requirements. In the diagnostic categories neurologic, musculoskeletal and psychiatric diseases the grounding rates went up. This cannot be attributed to more stringent health requirements, but possibly to the attitudes of pilots, who have become more critical of their working environment. PMID- 15550966 TI - [Treatment of malignant melanoma]. AB - The incidence and mortality of malignant melanoma has been steadily increasing over the past decades. The multidisciplinary Norwegian Melanoma Group published a set of guidelines for the treatment of cutaneous malignant melanoma in 1999. The guidelines were revised in 2002/2003. This article summarises the most important issues in the Norwegian guidelines and discusses some controversial aspects of melanoma management. PMID- 15550967 TI - [Blood transfusion and pretransfusion testing in patients with warm autoimmune haemolytic anaemia]. AB - BACKGROUND: In warm autoimmune haemolytic anaemia, patients have red cell autoantibodies that complicate the serological compatibility testing before transfusion. Different autoadsorption techniques are utilised for more extensive pretransfusion testing. We reviewed the literature on blood transfusion in these patients. We also present our experience with autoadsorption by papain and polyethylene glycol methods. MATERIAL AND METHODS: Blood samples from 13 patients with warm autoantibodies were analysed in parallel. After autoadsorption, antibody screening and identification tests were performed to detect possible red cell alloantibodies. If the serological testing is inadequate, clinically important alloantibodies can be ignored before transfusion, which can lead to haemolytic transfusion reactions. RESULTS: Autoantibodies were completely adsorbed in six of the 13 patients. In five, alloantibodies were detected. In three patients, different results were obtained with the two different techniques. INTERPRETATION: Restrictive transfusion practice in warm autoimmune haemolytic anaemia is recommended. Adequate pretransfusion testing in a specialised laboratory is required before transfusion. Autoadsorption with polyethylene glycol and papain can supplement each other in pretransfusion testing. PMID- 15550968 TI - [P-glycoprotein--a pump of significance for drug response]. PMID- 15550969 TI - [Ethical end-of-life decision making in nursing homes]. AB - BACKGROUND: A recent publication from Norwegian health authorities describes necessary routines for end-of-life decisions in hospitals. There are no comparable national recommendations regarding patients in nursing homes. 40% of deaths in Norway occur in nursing homes. METHODS: All nursing home physicians in Bergen received an open questionnaire on the practice of ethics, end-of-life decisions, and palliative care. RESULTS: 15 of the nursing homes physicians responded to the questionnaire, representing three quarters of the nursing homes and 1483 out of 1782 nursing home patients in Bergen (83%). Only two of institutions had written instructions for end-of-life decisions. Two thirds of the physicians considered use of morphine for the dying. Only a minority treated death rattle with scopolamine. 12 out of the 15 physicians administered life prolonging treatment with diuretics facing terminal pulmonary oedema, and 5 out of 15 administered antibiotics to those dying with pneumonia. 0.9% of the patients had a feeding tube. DISCUSSION: A majority of nursing home physicians have a high awareness of and willingness to give priority to end-of-life decisions, communication and palliative care. They ask for national recommendations in this challenging area. Unnecessary life-prolonging treatment and lack of palliative care are still major problems in Norwegian nursing homes. PMID- 15550970 TI - [Clinical use of haematopoietic stem cells from cord blood]. AB - A group of experts appointed by the Norwegian Centre for Health Technology Assessment (SMM) has undertaken a systematic review of available literature on the clinical effectiveness of transplanting haematopoietic stem cells from cord blood. A total of 17 studies form the documentary basis of the review. Autologous transplants of stem cells from cord blood have not been published. Retrospective studies suggest that the clinical effect of allogeneic cord blood transplants, at least in children, is comparable to transplants with allogeneic stem cells from bone marrow or peripheral blood cells. This review demonstrates the need for prospective studies comparing transplantations of cord blood with bone marrow or peripheral blood stem cells. PMID- 15550972 TI - [Documentation of violence and mortality in Darfur]. PMID- 15550973 TI - [Information about rights in connection with occupational injury and occupational disease]. PMID- 15550974 TI - [Routine ECG in methadone-assisted rehabilitation is wrong prioritization]. AB - The Norwegian Medicines Agency has recently reported dose-dependent QT prolongation and occurrence of Torsades des pointes in patients treated with methadone; the agency recommends that an ECG is taken before induction to methadone. We have performed a literature search in Medline and Embase. QT prolongation in methadone therapy is dose-dependent and primarily seen with doses higher than those usually used in maintenance therapy and/or in cases with known risk factors. Routine ECG should not be recommended, but before starting therapy the physician should secure an adequate case history including information on any family history of cardiac disease and other risk factors. ECG is recommended when doses higher than 130 mg/day are used or the serum concentration level exceeds 1200 nmol/l. QTc prolongation above 470 msek should be reported to the authors. PMID- 15550976 TI - [Thiazides (only) and hard endpoints]. PMID- 15550978 TI - [More about use of antibiotics in sepsis]. PMID- 15550979 TI - [Activity-based financing in psychiatry]. PMID- 15550980 TI - [Improved drug documentation]. PMID- 15550981 TI - [Innovation and re-creating should be encouraged]. PMID- 15550983 TI - Adaptive amplification and point mutation are independent mechanisms: evidence for various stress-inducible mutation mechanisms. AB - "Adaptive mutation" denotes a collection of processes in which cells respond to growth-limiting environments by producing compensatory mutants that grow well, apparently violating fundamental principles of evolution. In a well-studied model, starvation of stationary-phase lac(-)Escherichia coli cells on lactose medium induces Lac(+)revertants at higher frequencies than predicted by usual mutation models. These revertants carry either a compensatory frameshift mutation or a greater than 20-fold amplification of the leaky lac allele. A crucial distinction between alternative hypotheses for the mechanisms of adaptive mutation hinges on whether these amplification and frameshift mutation events are distinct, or whether amplification is a molecular intermediate, producing an intermediate cell type, in colonies on a pathway to frameshift mutation. The latter model allows the evolutionarily conservative idea of increased mutations (per cell) without increased mutation rate (by virtue of extra gene copies per cell), whereas the former requires an increase in mutation rate, potentially accelerating evolution. To resolve these models, we probed early events leading to rare adaptive mutations and report several results that show that amplification is not the precursor to frameshift mutation but rather is an independent adaptive outcome. (i) Using new high-resolution selection methods and stringent analysis of all cells in very young (micro)colonies (500-10,000 cells), we find that most mutant colonies contain no detectable lac-amplified cells, in contrast with previous reports. (ii) Analysis of nascent colonies, as young as the two-cell stage, revealed mutant Lac(+)cells with no lac-amplified cells present. (iii) Stringent colony-fate experiments show that microcolonies of lac amplified cells grow to form visible colonies of lac-amplified, not mutant, cells. (iv) Mutant cells do not overgrow lac-amplified cells in microcolonies fast enough to mask the lac-amplified cells. (v)lac-amplified cells are not SOS induced, as was proposed to explain elevated mutation in a sequential model. (vi) Amplification, and not frameshift mutation, requires DNA polymerase I, demonstrating that mutation is separable from amplification, and also illuminating the amplification mechanism. We conclude that amplification and mutation are independent outcomes of adaptive genetic change. We suggest that the availability of alternative pathways for genetic/evolutionary adaptation and clonal expansion under stress may be exploited during processes ranging from the evolution of drug resistance to cancer progression. PMID- 15550984 TI - State-dependent decisions cause apparent violations of rationality in animal choice. AB - Normative models of choice in economics and biology usually expect preferences to be consistent across contexts, or "rational" in economic language. Following a large body of literature reporting economically irrational behaviour in humans, breaches of rationality by animals have also been recently described. If proven systematic, these findings would challenge long-standing biological approaches to behavioural theorising, and suggest that cognitive processes similar to those claimed to cause irrationality in humans can also hinder optimality approaches to modelling animal preferences. Critical differences between human and animal experiments have not, however, been sufficiently acknowledged. While humans can be instructed conceptually about the choice problem, animals need to be trained by repeated exposure to all contingencies. This exposure often leads to differences in state between treatments, hence changing choices while preserving rationality. We report experiments with European starlings demonstrating that apparent breaches of rationality can result from state-dependence. We show that adding an inferior alternative to a choice set (a "decoy") affects choices, an effect previously interpreted as indicating irrationality. However, these effects appear and disappear depending on whether state differences between choice contexts are present or not. These results open the possibility that some expressions of maladaptive behaviour are due to oversights in the migration of ideas between economics and biology, and suggest that key differences between human and nonhuman research must be recognised if ideas are to safely travel between these fields. PMID- 15550985 TI - A chemoattractant role for NT-3 in proprioceptive axon guidance. AB - Neurotrophin-3 (NT-3) is required for proprioceptive neuron survival. Deletion of the proapoptotic gene Bax in NT-3 knockout mice rescues these neurons and allows for examination of their axon growth in the absence of NT-3 signaling. TrkC positive peripheral and central axons from dorsal root ganglia follow proper trajectories and arrive in close proximity to their targets but fail to innervate them. Peripherally, muscle spindles are absent and TrkC-positive axons do not enter their target muscles. Centrally, proprioceptive axons branch in ectopic regions of the spinal cord, even crossing the midline. In vitro assays reveal chemoattractant effects of NT-3 on dorsal root ganglion axons. Our results show that survival factor NT-3 acts as a short-distance axon guidance molecule for muscle sensory afferents as they approach their proper targets. PMID- 15550986 TI - DNA methylation profiling of the human major histocompatibility complex: a pilot study for the human epigenome project. AB - The Human Epigenome Project aims to identify, catalogue, and interpret genome wide DNA methylation phenomena. Occurring naturally on cytosine bases at cytosine guanine dinucleotides, DNA methylation is intimately involved in diverse biological processes and the aetiology of many diseases. Differentially methylated cytosines give rise to distinct profiles, thought to be specific for gene activity, tissue type, and disease state. The identification of such methylation variable positions will significantly improve our understanding of genome biology and our ability to diagnose disease. Here, we report the results of the pilot study for the Human Epigenome Project entailing the methylation analysis of the human major histocompatibility complex. This study involved the development of an integrated pipeline for high-throughput methylation analysis using bisulphite DNA sequencing, discovery of methylation variable positions, epigenotyping by matrix-assisted laser desorption/ionisation mass spectrometry, and development of an integrated public database available at http://www.epigenome.org. Our analysis of DNA methylation levels within the major histocompatibility complex, including regulatory exonic and intronic regions associated with 90 genes in multiple tissues and individuals, reveals a bimodal distribution of methylation profiles (i.e., the vast majority of the analysed regions were either hypo- or hypermethylated), tissue specificity, inter individual variation, and correlation with independent gene expression data. PMID- 15550987 TI - Lmo mutants reveal a novel role for circadian pacemaker neurons in cocaine induced behaviors. AB - Drosophila has been developed recently as a model system to investigate the molecular and neural mechanisms underlying responses to drugs of abuse. Genetic screens for mutants with altered drug-induced behaviors thus provide an unbiased approach to define novel molecules involved in the process. We identified mutations in the Drosophila LIM-only (LMO) gene, encoding a regulator of LIM homeodomain proteins, in a genetic screen for mutants with altered cocaine sensitivity. Reduced Lmo function increases behavioral responses to cocaine, while Lmo overexpression causes the opposite effect, reduced cocaine responsiveness. Expression of Lmo in the principal Drosophila circadian pacemaker cells, the PDF-expressing ventral lateral neurons (LN(v)s), is sufficient to confer normal cocaine sensitivity. Consistent with a role for Lmo in LN(v)function,Lmomutants also show defects in circadian rhythms of behavior. However, the role for LN(v)s in modulating cocaine responses is separable from their role as pacemaker neurons: ablation or functional silencing of the LN(v)s reduces cocaine sensitivity, while loss of the principal circadian neurotransmitter PDF has no effect. Together, these results reveal a novel role for Lmo in modulating acute cocaine sensitivity and circadian locomotor rhythmicity, and add to growing evidence that these behaviors are regulated by shared molecular mechanisms. The finding that the degree of cocaine responsiveness is controlled by the Drosophila pacemaker neurons provides a neuroanatomical basis for this overlap. We propose that Lmo controls the responsiveness of LN(v)s to cocaine, which in turn regulate the flies' behavioral sensitivity to the drug. PMID- 15550988 TI - The genetics of speciation by reinforcement. AB - Reinforcement occurs when natural selection strengthens behavioral discrimination to prevent costly interspecies matings, such as when matings produce sterile hybrids. This evolutionary process can complete speciation, thereby providing a direct link between Darwin's theory of natural selection and the origin of new species. Here, by examining a case of speciation by reinforcement in Drosophila,we present the first high-resolution genetic study of variation within species for female mating discrimination that is enhanced by natural selection. We show that reinforced mating discrimination is inherited as a dominant trait, exhibits variability within species, and may be influenced by a known set of candidate genes involved in olfaction. Our results show that the genetics of reinforced mating discrimination is different from the genetics of mating discrimination between species, suggesting that overall mating discrimination might be a composite phenomenon, which in Drosophila could involve both auditory and olfactory cues. Examining the genetics of reinforcement provides a unique opportunity for both understanding the origin of new species in the face of gene flow and identifying the genetic basis of adaptive female species preferences, two major gaps in our understanding of speciation. PMID- 15550990 TI - Treatment of postherpetic neuralgia. AB - Postherpetic neuralgia (PHN) is a serious complication of herpes zoster that has a predilection for older individuals. PHN is often associated with significant morbidity, and it can cause insomnia, fatigue, depression and interference with daily activities in affected individuals. Treatment for PHN is initiated with antivirals during the acute herpes zoster outbreak. Acyclovir (Zoviraxr, GlaxoSmithKline), valacyclovir (Valtrex, GlaxoSmithKline) or famciclovir (Famvir, Novartis) can be used to treat herpes zoster, and all three have been shown to reduce the duration of the herpetic rash and zoster-associated pain. These antivirals are most effective when used within the first 72 hours of the onset of the rash. Side-effects of these antivirals are low and include nausea, vomiting, abdominal pain and headache. Other treatment options for PHN include topical analgesics, opioid analgesics, tricyclic antidepressants and gabapentin. Because of the complexity of PHN, most patients require a combination of treatment modalities for adequate pain relief. PMID- 15550991 TI - The use of lasers and intense pulsed light sources for the treatment of pigmentary lesions. AB - Lasers and intense pulsed light sources are frequently used for the treatment of pigmented lesions, and the appropriate selection of devices for different lesions is vital to achieving satisfactory clinical outcomes. In dark-skinned patients, the risk of post-inflammatory hyperpigmentation is of particular importance. In general, long-pulse laser and intense pulsed light sources can be effective with a low risk of post-inflammatory hyperpigmentation (PIH) when used for the treatment of lentigines. However, for dermal pigmentation and tattoo, Q-switched lasers are effective, with a lower risk of complications. In the removal of melanocytic nevi, a combined approach with a long-pulse pigmented laser and a Q switched laser is particularly applicable. PMID- 15550992 TI - Tacrolimus ointment (Protopic) for atopic dermatitis. AB - Tacrolimus ointment (Protopic, Fujisawa) is an effective agent in a class of topical immunomodulators. Its mechanism of action is based on calcineurin inhibition, which results in decreased T-cell activation and inflammatory cytokine release. Tacrolimus ointment is safe and effective for short- and long term treatment of atopic dermatitis (AD) in pediatric and adult patients. The most common adverse events associated with its use are a transient burning sensation and pruritus at the site of application. Unlike topical corticosteroid agents, tacrolimus ointment does not cause a reduction in collagen synthesis or skin thickness. Because tacrolimus ointment does not cause skin atrophy, it may be safely used for months or years on all skin areas, including the face and intertriginous areas. PMID- 15550993 TI - Laser treatment of leg veins. AB - The role of lasers and intense pulsed light sources has gained increasing popularity in the management of both cosmetic telangiectasias and medically significant symptomatic varicose vein disease. These advances include endovascular technologies, novel cooling technologies, variable spot sizes and pulse durations, as well as the ability to deliver high-energy fluences. These advances have allowed the delivery of sufficient energy allowing more efficient pan-endothelial necrosis without affecting epidermal structures, and yielding a lower complication profile such as post-inflammatory hyperpigmentation and epidermal surface irregularities. The advent of extended-pulse, longer wavelength technologies such as the 1064 Neodymium:Yttrium Aluminum Garnet (Nd:YAG) laser have allowed the treatment of individuals with darker skin phenotypes, as well as treatment of deep blue reticular veins up to 3 mm in diameter in a monomodal fashion. Combined approaches of sclerotherapy plus laser treatments performed during the same treatment session may produce synergistic results in selected individuals. PMID- 15550994 TI - The interactions between human dendritic cells and microbes; possible clinical applications of dendritic cells. AB - The dendritic cells comprise several subsets that induce and regulate the immune responses against foreign and self-antigens, and that can therefore function as initiators of protective immunity and inducers of central or peripheral tolerance. The different subpopulations of dendritic cells interact with and also influence other cell populations of the immune system, such as T and B lymphocytes and natural killer cells. The factors that determine the given dendritic cell functions depend on the state of maturation and the local microenvironment. The interactions between dendritic cells and microorganisms are rather complex, but progress in the past few years has shed light on several aspects of these interactions. This review lays emphasis on the interactions between human dendritic cells, important components of the intima of arterial specimens at areas predisposed to atherosclerotic lesions, and Chlamydia pneumoniae and cytomegalovirus, the human pathogens most strongly implicated in the development of atherosclerosis. In addition, several examples of the potential clinical applications of dendritic cells are described. PMID- 15550995 TI - Understanding human dendritic cell biology through gene profiling. AB - Dendritic cells are potent antigen presenting cells whose function has been associated with a variety of immunological disorders. Because of their relevance to human disease, extensive efforts have been made to gain a better understanding of their biology. One aspect of these efforts has been in the identification of pertinent molecules expressed in these cells through gene profiling experiments and proteomics. In this review, we summarize the results from the various profiling studies that have been done with human dendritic cells. We focus on molecules, which have been confirmed by other methods, such as quantitative PCR, or have been identified in multiple profiling studies to be expressed in the respective dendritic cell type. Through such profiling experiments and subsequent analysis, interesting molecules have been identified which can be further studied to determine their role in dendritic cell biology. PMID- 15550989 TI - Distinct genomic integration of MLV and SIV vectors in primate hematopoietic stem and progenitor cells. AB - Murine leukemia virus (MLV)-derived vectors are widely used for hematopoietic stem cell (HSC) gene transfer, but lentiviral vectors such as the simian immunodeficiency virus (SIV) may allow higher efficiency transfer and better expression. Recent studies in cell lines have challenged the notion that retroviruses and retroviral vectors integrate randomly into their host genome. Medical applications using these vectors are aimed at HSCs, and thus large-scale comprehensive analysis of MLV and SIV integration in long-term repopulating HSCs is crucial to help develop improved integrating vectors. We studied integration sites in HSCs of rhesus monkeys that had been transplanted 6 mo to 6 y prior with MLV- or SIV-transduced CD34(+)cells. Unique MLV (491) and SIV (501) insertions were compared to a set of in silico-generated random integration sites. While MLV integrants were located predominantly around transcription start sites, SIV integrants strongly favored transcription units and gene-dense regions of the genome. These integration patterns suggest different mechanisms for integration as well as distinct safety implications for MLV versus SIV vectors. PMID- 15550996 TI - Chronic blockade of nitric oxide biosynthesis in rats: effect on leukocyte endothelial interaction and on leukocyte recruitment. AB - INTRODUCTION: Previous studies showed that animals chronically treated with NG nitro-L-arginine methyl ester (L-NAME) have a reduced inflammatory reaction. Now the role of L-NAME treatment (20 mg/Kg/day/14 days) on leukocyte mobilisation was assessed in rats. METHODS: In vivo leukocyte recruitment evoked by Bothrops jararaca venom (BjV) and nitrite/nitrate (NO2-/NO3-; Griess reaction) were evaluated in the air pouch cavity. Haematological parameters were evaluated in the bone marrow and in the peripheral compartment. Microcirculatory blood flow, number of rolling and adhered leukocytes, vascular reactivity and mast cell activity were studied by intravital microscopy. Blood pressure was measured by the tail-cuff method. L-selectin and beta(2) integrin expressions on peripheral and bone marrow leukocytes were quantified by flow cytometry. RESULTS: When compared with control rats (D-NAME) L-NAME treated rats had reduced PMN cell infiltrate (50%) and NO2-/NO3- (27%) in the air pouch cavity. Rolling leukocytes were decreased (70%) in L-NAME-treated animals, which was reversed by topical application of NO donor (SIN-1). BjV stimulation increased the number of rolling and adhered leukocytes only in control rats. Systemic blood pressure, microcirculatory blood flow and microvascular reactivity was not altered by the treatment. Only the vessel response to acetylcholine was delayed in treated rats. Peripheral PMN cells were increased by L-NAME treatment (100%), but the number of bone marrow cells was not altered. The treatment reduced L-selectin expression on circulating leukocytes, by either with (16%) or without (26%) stimulation with BjV; PMN cells were more affected (32-37%). Impairment of L-selectin expression was also verified in bone marrow cells under stimulation with BjV. CONCLUSIONS: Results show that this schedule of L-NAME treatment promotes a decrease on L selectin expression. This effect may promote the standstill of leukocytes in the blood compartment and may be responsible, at least in part, for the observed deficient leukocyte-endothelium interactions with subsequent impairment of leukocyte migration to the inflammatory site. PMID- 15550997 TI - Spontaneous histamine secretion during isolation of rat cardiac mast cells. AB - OBJECTIVE: To test the hypothesis that spontaneous release of histamine occurring during an isolation protocol may modify responses of rat cardiac mast cells (connective tissue-type mast cells) to secretagogues. METHODS: We assessed two protocols for enzymatic dispersion utilizing collagenase, hyaluronidase, and deoxyribonuclease; with protease (Protocol 1, n = 8) or without protease (Protocol 2, n = 3). Spontaneous release of histamine was quantified following mechanical and enzymatic dispersion of the whole heart. RESULTS: Total histamine loss (Mean +/- SEM) was 963+/-92 and 833+/-60 ng/g of tissue weight following Protocols 1 and 2. Percentages of histamine release from cell isolates following Protocol 1 were 40+/-5%, 41+/-6%, and 51+/-7% at 0, 30, and 300 microg/mL of compound 48/80. CONCLUSIONS: Enzymatic dispersion of cardiac mast cells affects their response to secretagogues. PMID- 15550998 TI - Spontaneous aggregation of leukocytes in active ulcerative colitis might be ICAM 1 dependent. AB - OBJECTIVE: In active stages of ulcerative colitis (UC), a tendency for neutrophils to aggregate in the colonic lamina propria is mediated by yet unidentified surface receptors. The aim was to assess the spontaneous leukocyte aggregation and the aggregation induced by bacteria-derived products in UC and to evaluate the involvement of ICAM-1 and beta(2)-integrins in this aggregation. MATERIALS AND METHODS: Blood was drawn from 10 patients with quiescent UC, 10 patients with active UC, and 10 healthy volunteers. The blood was stimulated with LPS or fMLP with subsequent blocking of CD11b or ICAM-1 with specific antibodies. The aggregation was assessed on glass slides with an automated image analyzer (Inflamet). RESULTS: The spontaneous leukocyte aggregation was increased in quiescent and active UC as compared to healthy controls (p < 0.05). Although not statistically significant, LPS and fMLP seemed to increase the leukocyte adhesiveness, and also a tendency towards inhibition of the leukocyte aggregation was observed by blocking ICAM-1. CONCLUSIONS: Increased adhesiveness of circulating leukocytes seems to be involved in the pathogenesis of UC, and ICAM-1 is suggested to be a part of this phenomenon. The results indicate an altered basic neutrophil response in UC. PMID- 15550999 TI - An orally active matrix metalloproteinase inhibitor, ONO-4817, reduces dextran sulfate sodium-induced colitis in mice. AB - OBJECTIVE: Over-expression of matrix metalloproteinases (MMPs) can accelerate tissue destruction and disrupt subsequent tissue repair. A dextran sulfate sodium (DSS) colitis model was established to examine the effects of MMP inhibition, by an orally active MMP inhibitor ONO-4847, on colonic inflammation. MATERIALS AND METHODS: Acute colitis was induced in female BALB/c mice by giving 8% DSS orally in drinking water for 7 days. The animals were randomized into groups receiving different concentrations of ONO-4847 or vehicle by oral gavage every day. mRNA levels of 4 MMPs and a tissue inhibitor of MMP (TIMP-1) were measured by RT-PCR in intestinal tissue isolated from mice after DSS administration. Colonic mucosal injury and inflammation were evaluated clinically, biochemically, and histologically. The clinical disease activity index (DAI), including body weight loss, stool consistency, and blood in feces, was examined. Moreover, mucosal tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma were determined by immunoassay. RESULTS: The intestinal expression of MMP-3, -7, 9, and -12 and TIMP 1 mRNA was upregulated after DSS administration. Shortening of the colon was significantly reversed by ONO-4847 at a dose of 30 mg/kg. DAI in DSS-treated mice was significantly lower in the ONO-4847-treated mice compared with the control mice. Histological study also showed a reduced infiltration of inflammatory cells, especially neutrophils, and reducedmucosal cell disruption in ONO-4847 treated mice compared with the control mice. The increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by co-administration with ONO 4847. ONO-4847 also inhibited increases in the mucosal TNF-alpha and IFN-gamma content after DSS administration. CONCLUSION: Improvements in DSS colitis in response to ONO-4847 suggest that activation of MMPs contributes to the initiation/amplification of colonic inflammatory injury by mechanisms including oxidative damage as well as enhancement of inflammatory cytokine release. PMID- 15551000 TI - Inflammatory cytokine levels in paw tissues during development of rat collagen induced arthritis: effect of FK506, an inhibitor of T cell activation. AB - OBJECTIVE AND DESIGN: To characterize rat collagen-induced arthritis (CIA) on the basis of levels of inflammatory cytokines, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 in paw tissues, and further investigate the effect of FK506 (tacrolimus), a potent inhibitor of T cell activation, on cytokine levels. METHODS: CIA was induced in female Lewis rats. The volume of hindpaws was measured before and after collagen immunization. TNF-alpha, IL-1beta and IL-6 levels in paw tissue extracts were determined by ELISA. Proteoglycan contents of cartilage in femoral heads was measured as an indication of cartilage destruction. To assess the effect of FK506 on inflammatory cytokine levels, rats were orally treated with 5 mg/kg of FK506 from days 14-21. RESULTS: TNF-alpha a level in paw tissues did not significantly change compared to levels found before collagen immunization, throughout development of CIA. In contrast, IL-1beta and IL-6 levels in paw tissues significantly increased between day 14 and day 28 after collagen imuninization, when the arthritis was at a developed stage. Therapeutic treatment with FK506 reduced the elevated level of IL-6, but not IL 1beta, in paw tissue. FK506 treatment was effective in suppressing paw swelling and also recovering the loss of proteoglycan contents in the cartilage. CONCLUSIONS: Levels of IL-1beta and IL-6, but not TNF-alpha , in paw tissue were upregulated in association with the development of arthritis in rat CIA. These results suggest that IL-1beta and IL-6, rather than TNF-alpha , may play important roles at local inflammatory sites in producing joint destruction in rat CIA. FK506 may improve arthritis in established stages of CIA, by reducing the elevated level of IL-6. PMID- 15551001 TI - Diffusible signal to murine alveolar macrophages from lipopolysaccharide- and Escherichia coli-stimulated lung Type II epithelial cells. AB - OBJECTIVE AND DESIGN: To demonstrate a diffusible intercellular macrophage activation factor secreted by Type II alveolar epithelial cells (AECs) in transwell co-cultures. MATERIALS: T(7), our Type II conditionally immortalized AEC line; MH-S, an alveolar macrophage cell line; Lipopolysaccharide (LPS) or uv killed Escherichia coli (UVEC) for antigen presentation. METHODS: LPS or UVEC stimulation of T(7) cells in the lower chamber was investigated for ability to activate MH-S cells in the upper chamber, as assayed by nitric oxide production and western blots for inducible nitric oxide synthase-2. RESULTS: Both transwell and UVEC-conditioned medium experiments showed secretion of an MH-S activation factor by T(7) cells. Many common inflammatory cytokines were ruled out as this immunoactivator. CONCLUSION: Demonstration of a diffusible activation factor produced by Type II AECs supports their potential role as first responders of innate immunity in the lung. PMID- 15551002 TI - Single nucleotide polymorphisms and haplotypes of histamine N-methyltransferase in patients with gastric ulcer. AB - INTRODUCTION: Histamine plays a crucial role in the regulation of gastric acid secretion, which is involved in the pathogenesis of peptic ulcer. Histamine N methyltransferase (HNMT) is the major metabolizing enzyme for histamine inactivation in human stomach. OBJECTIVE: This study aims to determine whether there exists a relationship between HNMT gene polymorphisms and the risk for gastric ulcer (GU). METHODS: 118 GU patients and 154 ethnically matched control subjects were enrolled and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assays were developed to genotype all these subjects for the T-1637C, C-411T, C314T and A1097T point mutations in HNMT gene. Haplotypes were reconstructed from the genotype data. RESULTS: Frequencies of the variant alleles in cases and controls were 0.398 vs 0.396 for T-1637C, 0.144 vs 0.110 for C-411T, 0.034 vs 0.042 for C314T, and 0.242 vs 0.273 for A1097T, respectively, with no significant difference for any locus between the two groups (all P > 0.05). Also the frequencies of genotypes, haplotypes and haplotype pairs based on these polymorphisms did not differ significantly between cases and controls. CONCLUSION: This study provided no evidence for the involvement of HNMT polymorphisms in the susceptibility to GU. PMID- 15551003 TI - Cockroach allergen extract stimulates protease-activated receptor-2 (PAR-2) expressed in mouse lung fibroblast. AB - OBJECTIVE: To investigate whether cockroach allergen extract can stimulate Protease-activated receptor 2 (PAR-2) expressed in mouse lung fibroblast. MATERIALS: We established an immortalized lung fibroblast cell line, DM5, from PAR-2 deficient mice. By stable transfection with either an empty vector (DM5/EV) or an expression vector encoding mouse PAR-2 cDNA (DM5/Par2), a pair of lung fibroblast cell lines with or without functional PAR-2 expression were prepared. TREATMENT: The cells were exposed to cockroach allergen extract [up to 800 protein nitrogen unit (PNU)/ml], trypsin (up to 100 nM), SLIGRL agonist peptide (up to 500 microM), and trans-cinnamoyl-LIGRO agonist peptide (up to 400 microM). METHODS: The cells were loaded with Fluo-3 calcium indicator and mobilization of intracellular calcium with the stimuli was monitored by a fluorometric plate reader. Extracellular signal-regulated kinase (ERK) phosphorylation was examined by Western blot analysis using an anti-phospho ERK antibody. RESULTS: The cockroach extract induced intracellular calcium transients in a concentration dependent manner in DM5/Par2 but not in DM5/EV. The activity was abolished when the cockroach extract was heat denatured or pre-incubated with PMSF (phenylmethanesulfonyl fluoride) prior to the assay. Concomitantly, ERK phosphorylation was seen in DM5/Par2 with the cockroach extract but not with a heat-denatured extract. The responses were sensitive to an inositol-1,4,5 triphosphate antagonist (2-APB) indicating that calcium was mobilized from intracellular store. CONCLUSIONS: Cockroach allergen extract can activate PAR-2 and thereby stimulate mouse lung fibroblasts likely through protease(s). The present study proposes a potential mechanism for cockroach antigens, similar to house dust mite antigens, in the etiology of respiratory diseases. PMID- 15551006 TI - [Nononcologic abdominal surgery in the elderly]. AB - Due to rising life expectancy of the population, the proportion of elderly patients requiring surgery is rising as well. Present aspects of selected, typical, nononcologic diseases of elderly people are discussed. The key to success in their treatment is not to consider primarily the patient's calendrical age but to assess the individual profile of risk factors on the basis of comorbidities. The kind, extent, and timing of an operation has to be based on this assessment. For the treatment of acute diseases (e.g., complicated ulcera, cholecystitis, appendicitis, mesenteric ischemia, and diverticulitis), a rapid and efficacious diagnostic algorithm is essential that takes into account the reduced functional reserve of old people. Constructive interdisciplinary cooperation and minimally invasive techniques play dominant roles in both diagnosis and therapy. Given these prerequisites, there is no reason to withhold surgical intervention from elderly patients. PMID- 15551007 TI - [Needs of health insurers regarding surgery for the aged in Germany]. AB - The increasing proportion of elderly persons presents one of the most urgent problems to European societies and has a decisive effect on social insurance systems. Costs are rising rapidly due to longer life and the falling number of births. Health care is especially hard-hit because of medical progress and growing numbers of procedures during old age. Despite these problems, the German health care system is still effective. In contrast to neighboring European states, there are neither waiting lists nor rationing of services. With respect to financing, future developments cannot be predicted. Planning scenarios taking enormously higher costs into account often result in demands for rationing, which the German health insurance companies do not accept. This study examines economic figures from the largest German health insurer and attempts to draw a picture of future health care for old people. New possibilities for prevention and resourcing and the need for rationing are discussed. The first, promising projects in these respects have already been started, with the aim of better health and quality of life for the elderly. PMID- 15551005 TI - [Thoracic surgery in the elderly]. AB - In the near future, over 40% of patients with lung cancer will be over 70 years old at the time their disease is diagnosed. Age per se, however, should not lead to the denial of a potentially curative surgical intervention. It has been shown that older patients (over 70 years), as well as patients over 80 years of age, may tolerate a lobectomy or even a pneumonectomy quite well. Most patients with lung cancer are present or former smokers and have underlying pulmonary problems, especially chronic obstructive lung disease. They are at high risk of both morbidity and mortality from surgery due to significant cardiovascular disease. The indications for surgical intervention should be based on reliable preoperative tumor staging and pulmonary assessment by an experienced interdisciplinary panel of physicians, taking into consideration the individual cardiopulmonary status of the patient. This assessment, combined with the American Society of Anesthesiologists risk classification and the overall clinical assessment by the surgeon, will provide the best available evidence for carefully weighing the benefits and risks of an operation. The responsibility for this assessment must be viewed-in the case of early stage lung cancer-in relation to the relative lack of alternative treatments for surgical intervention with comparable 5-year survival rates (>50%). PMID- 15551008 TI - ["FRAGILE"-Old people and surgery]. AB - As life expectancy and modern surgical and intensive care techniques develop, the number of old patients in surgery is rising. Associated are problems with the indication for surgical intervention and rising incidence of "typical" peri- and postoperative complications such as postoperative delirium. Geriatric assessment serves to identify patients at risk of developing complications, to describe the postoperative course of functional and cognitive abilities, and to adjust therapeutic strategies to the individual needs of these patients. The most important instruments of geriatric assessment are described. Postoperative delirium, the most prevalent complication in surgical geriatric patients, is discussed in detail with regard to risk factors, prevalence, diagnosis, and therapeutic options. PMID- 15551009 TI - [Population developments in Germany until 2050]. AB - While the total population in Germany is expected to diminish until 2050, the proportion of old and very old persons will rise considerably. The annual frequency of sicknesses, clinic days, and fatalities of old persons will likewise strongly increase. This raises difficult problems for the future design of the German public health care system. In any case, reforms will contain elements of rationing which will lead to a less expansion of the field of activity for geriatric physicians and surgeons than the purely demographic development would suggest. This dampening effect of rationing reforms on geriatric medical treatment and surgery, however, might possibly be least when done in an age specific way and combined with long-term individual provisions through savings and health insurance. PMID- 15551010 TI - [Quality of life after subtotal resection and gastrectomy for gastric cancer]. AB - Recently, quality of life has become a main objective in surgical therapy. Apart from the oncological consequences, the quality of results after gastric carcinoma resection are mainly determined by social and psychological aspects of life, early postoperative complications, and long-term nutritive/functional parameters. Of 338 patients who underwent gastric resection for adenocarcinoma of the stomach, quality of life was assessed in 73 recurrence-free patients by means of the Gastrointestinal Life Quality Index (GLQI). The median age was 71.9 years, and surgery had been carried out at least 1 year prior to the investigation (median follow-up 4.5 years). Patients with subtotal gastric resection displayed significantly higher GLQI scores (120 [97-138] points) than patients with gastrectomy (116 [70-139] points;p=0.047). Among partial parameters of the life quality index, physical functions were significantly better after subtotal resection (p=0.040), while the emotional status (p=0.147) and social activities (p=0.337) did not differ between the two groups. Abdominal symptoms (p=0.081) and the nutritional function (p=0.228) were insignificantly different. The number of meals (4 vs. 5 meals per day) and the loss of weight since surgery (5 vs. 10 kg) were less after subtotal resection than after gastrectomy. However, the latter parameter did not reach statistical significance. PMID- 15551011 TI - [Increasing abdominal pain caused by abdominal tumor in a 21-year-old female]. AB - Lymphangiomas are congenital malformations which occur mostly in the neck and head region of children. They are rarely located in the mesenterium or retroperitoneum in adults. We present a case of mesenteric cystic lymphangioma in a young woman who was admitted to hospital with a history of increasing abdominal pain. We discuss diagnostic means, differential diagnosis, and therapeutic management of the disease. PMID- 15551012 TI - [Thoracic injuries associated with acute traumatic paraplegia of the upper and middle thoracic spine]. AB - AIM: The aim of this study was to analyse the number of thoracic injuries associated with acute traumatic paraplegia of the upper and middle thoracic spine and review the early management in respect to treatment standards. METHODS: Using a prospective study protocol, data were sampled and analyzed from 12 December 2000 to 31 March 2002 at a level 1 trauma center. RESULTS: Twenty-two consecutive patients were included in the study. Sixteen suffered severe chest traumata. Lung contusion was diagnosed in 81%, followed by haemopneumothorax (75%) and fracture of the bony chest (75%). Intubation was performed 12 times in all. Chest drainage was performed in 14 patients. The mean duration of artificial ventilation was 20 days (range 2-93) and of intensive care treatment 25 days (range 2-93). Five patients died. CONCLUSIONS: Acute traumatic paraplegia of the upper and middle thoracic spine caused by high energy trauma is highly associated with severe chest trauma. Therefore, respiratory impairment must be kept in mind during the early treatment. If respiratory failure becomes more evident, emergency procedures such as intubation and chest drainage have to be performed. Secondary transfer should be avoided. PMID- 15551013 TI - [Contraction of the hip and fecal drainage via a fistula tract 30 years after "appendectomy"]. AB - A 46-year-old female was admitted with increasing fecal drainage via a fistula tract in the right inguinal region. She had a history of surgery for appendicitis 30 years previously, from which there was disturbed wound healing resulting in a blunt fistula, and the patient suffered from contraction of the right hip. Computed tomographic scan and ultrasound demonstrated an inflammatory mass in the right inguinal region. Colonoscopy demonstrated a stenosis of the rectosigmoid junction but did not provide any further specific information. Surgery revealed the presumed diagnosis of complicated Crohn's disease, but an advanced squamous cell carcinoma was also identified. The patient died 23 months later due to generalized tumor. Although malignant transformation of a fistula tract is rare, this case demonstrates that long-standing fistulas should be cured as far as possible without significant morbidity. In the case of incurable fistulas, malignancy must definitely be excluded if the clinical appearance of the fistula changes. PMID- 15551014 TI - [Malignant melanoma of the uvula and soft palate]. AB - Malignant melanoma of the uvula and soft palate is very rare. We describe a case demonstrating the typical aggressive behavior of mucosal melanoma in contrast to cutaneous melanoma. Regional lymph node and distant metastases often develop rapidly in this type of melanoma. Despite aggressive surgical approaches, other treatment options are often limited and usually palliative. Therefore, early diagnosis is critical in patients with mucosal melanoma. PMID- 15551015 TI - [Sunlight-induced granulomatous reaction to permanent lip liner]. AB - The demand for permanent make up and especially permanent lip liner has increased in recent years in Germany. Information about complications is not available in the medical literature, because tattooing is a trade whose results and problems are not monitored and documented, as is the case for medical procedures. A female patient with permanent lip liner experienced no complications for one year. Then, after intensive sun exposure she developed solid, linear papules along the outer margin of her lips. Histology revealed a granulomatous tissue reaction with a marked lymphohistiocytic infiltrate including epithelioid histiocytes. Treatment with topical steroids over three weeks led to a complete healing without relapse. To the best of our knowledge this is the first report on UV-light-induced granulomatous reaction to permanent lip liner. PMID- 15551017 TI - [The effects of recurrent otitis media with effusion on speech development]. AB - Otitis media with effusion (OME) may have adverse effects on children's cognitive, language, speech and psychosocial development. This has been demonstrated using several lines of research, the results of which, however, are not unequivocal. The question of how OME can interact with language, speech and cognitive development is also a subject of debate. According to the acoustic phonetic theory, reduced perceptual ability during episodes of OME results in a degraded representation of subphonemic features of speech sounds, leading to speech delay. The mediated theory holds that fluctuant hearing loss endangers diffuse cognitive-linguistic effects, affecting speech perception and production. PMID- 15551018 TI - [Realistic early and late results after otosclerosis surgery and presentation of a technique involving almost no complications]. AB - Although the modern technique of otosclerosis surgery introduced by John Shea on 1st May 1956 has already been used throughout the world for almost fifty years it has not been possible to bring in line the differing opinions of surgeons concerning the optimum operation technique, the problems of the fenestration of the footplate, material and form of stapes replacement prosthetics and kind and place of their anchoring on the incus. Moreover, there is considerable disagreement among surgeons on a realistic evaluation of the findings of early and late results. During the last few decades otosclerosis surgery has become the hallmark of modern ear surgery and has been practiced in very highly specialized departments as well as in outpatient departments. The author, who has been very active in otosclerosis surgery since 1959-since 1979 with a modified personal technique-, wishes to prove that it is both meaningful and essential to modify the operation technique further, which is based on his own experiences and on more than 100 international publications. In this paper he presents his modified method, which has been tested on 1800 ears since 1979, as well as the results, achieved in this way. PMID- 15551019 TI - [Computer-assisted surgery (CAS) for the optimal treatment of craniofacial fibrous dysplasia]. AB - BACKGROUND AND OBJECTIVE: The rare bone disease craniofacial fibrous dysplasia is only treated in cases of visible deformities of the splanchnocranium and neurocranium, compression syndromes, and delocation of nasal and paranasal drainage. METHODS AND PATIENTS: In a retrospective quality assessment analysis of six patients with craniofacial monostotic fibrous dysplasia, the indications were analysed for situations in which computer-assisted surgery (CAS) might be helpful. RESULTS AND CONCLUSIONS: CAS serves as an intraoperative basis for the assessment of cosmetic-aesthetic corrections of visible bone deformities of the splanchnocranium and neurocranium, and allows the controlled removal of pathologically affected bone in contralateral symmetry. This seems to be important, especially at the anterior skull base, as deformities of the human face produce strong psychological suffering. Furthermore, CAS allows the configuration of implants for defect reconstruction after focal restorations with bony radical surgery. Implants of various material can be prefabricated from these data, which can then be exactly fitted to cover the defect. PMID- 15551020 TI - [Prolonged healing process after endonasal nasal sinus surgery. Gastroesophageal reflux as a cause?]. AB - BACKGROUND: The healing process after functional endoscopic sinus surgery (FESS) normally takes weeks to months under an intensive therapy regime. Occasionally, symptomatic failures and local problems occur despite extensive follow-up treatment. PATIENTS AND METHODS: Over a 3-year period 900 patients underwent FESS in our department. Of these patients, 79 were found with no tendency toward improvement of symptoms and signs more than 3 weeks after the operation. Nasal obstruction, frontal headache, and postnasal drainage were evaluated to be the frequent postoperative complaints. Scarring of the middle meatal antrotomy and the ethmoid, inflammatory granulation, and early recurrence of polyps occurred as the local problems. Historical factors and clinical signs of gastroesophageal or laryngopharyngeal reflux were examined in this group. Afterwards, proton pump inhibitors were prescribed on a trial basis. RESULTS: Historical clues of reflux were reported by 30 patients only. However, clinical signs of a reflux were found in 66 cases. Rhinological symptoms and signs were improved in 60 patients following the application of proton pump inhibitors over 2 weeks. CONCLUSION: Our results support earlier experiences reported in the literature. Laryngopharyngeal reflux could be a factor altering the healing process after FESS. PMID- 15551021 TI - ["State of the HAART". Current strategies for antiretroviral therapy]. AB - The goal of antiretroviral therapy is to interrupt the disease progression and to approach normal life expectancy for individuals with HIV. The condition for this is to deter the emergence of resistant HIV. This aim is achieved by improved patient drug adherence, sufficient enteral resorption and the control of interactions, as well as good tolerability and full antiviral activity. An insufficient drug exposure for protease inhibitors is overcome by the use of a booster agent. Emerging HIV-resistance can be described in a phase model:Phase 1:replication of wild-type HIV under incomplete suppressive therapy. Phase 2:transformation of wild-type into resistant HIV through accumulation of mutations. A preventive, resistance-guided switch of therapy preserves therapeutic options for the future. Phase 3:replication of resistant HIV (no more mutations possible).a)stable immunity: continue with failing therapy.b)threatening disease progression: resistance-guided switch to a salvage regimen. PMID- 15551022 TI - [Carotid endarterectomy in patients without neurological symptoms]. PMID- 15551023 TI - Meta-analysis of clinical characteristics of 299 carriers of LMNA gene mutations: do lamin A/C mutations portend a high risk of sudden death? AB - This study evaluated common clinical characteristics of patients with lamin A/C gene mutations that cause either isolated dilated cardiomyopathy or dilated cardiomyopathy in association with skeletal muscular dystrophy. We pooled clinical data of all published carriers of lamin A/C gene mutations as cause of skeletal and/or cardiac muscle disease and reviewed ECG findings. Cardiac dysrhythmias were reported in 92% of patients after the age of 30 years; heart failure was reported in 64% after the age of 50. Sudden death was the most frequently reported mode of death (46%) in both the cardiac and the neuromuscular phenotype. Carriers of lamin A/C gene mutations often received a pacemaker (28%). However, this intervention did not alter the rate of sudden death. Review of the ECG findings typically showed a low amplitude P wave and prolongation of the PR interval with a narrow QRS complex. This meta-analysis suggests that cardiomyopathy due to lamin A/C gene mutations portends a high risk of sudden death, and that this risk does not differ between subjects with predominantly cardiac or neuromuscular disease. This implies then that all carriers of a lamin A/C gene mutation need to be carefully screened with particular emphasis also on tachyarrhythmias. Prospective studies are needed to evaluate risk stratification and proper treatment strategies. PMID- 15551024 TI - Increased levels of adenine nucleotide translocator 1 protein and response to oxidative stress are early events in facioscapulohumeral muscular dystrophy muscle. AB - Facioscapulohumeral muscular dystrophy (FSHD), an autosomal dominant neuromuscular disorder, has been causally related to deletion of tandemly arrayed 3.3 kb repeats (D4Z4) on chromosome 4q35. Although increased expression of several 4q35 genes has been reported, two recent studies dispute this, finding no significant changes in the transcriptional level of any of the 4q35 genes, among which is the heart and muscle-specific isoform of the adenine nucleotide translocator (ANT1). We found markedly increased levels of ANT1 protein in both unaffected and affected FSHD muscles in comparison to control healthy muscles. Comparative protein expression analysis between healthy, Duchenne muscular dystrophy, and FSHD muscle shows that proteins involved in mitochondrial function and protection from oxidative stress are also reproducibly and specifically modified in all FSHD muscles, including clinically unaffected muscles. Increased ANT1 expression and mitochondrial dysfunction may thus be initial events in FSHD pathogenesis and represent potential therapeutic targets. PMID- 15551025 TI - Extinction may not be forever. AB - Here we review the phenomenon of ecomorph evolution and the hypothesis of iterative climatic cycles. Although a widely known phenomenon, convergent evolution has been underappreciated in both its scope and commonality. The power of natural selection to override genealogy to create similar morphologies (even among distantly related organisms) supports classical Darwinian evolution. That this occurs repeatedly in stratigraphically closely spaced intervals is one of the most striking features of Earth history. Periodic extinctions followed by re evolution of adaptive types (ecomorphs) are not isolated occurrences but are embedded within complex ecological systems that evolve, become extinct, and repeat themselves in temporal synchrony. These complexes of radiation and extinction bundle the biostratigraphic record and provide the basis for a global stratigraphy. At this scale, climatic change is the only mechanism adequate to explain the observed record of repeating faunas and floras. Understanding of the underlying causes may lead to predictive theories of global biostratigraphy, evolutionary processes, and climatic change. PMID- 15551026 TI - Ant-plant-herbivore interactions in the neotropical cerrado savanna. AB - The Brazilian cerrado savanna covers nearly 2 million km2 and has a high incidence on foliage of various liquid food sources such as extrafloral nectar and insect exudates. These liquid rewards generate intense ant activity on cerrado foliage, making ant-plant-herbivore interactions especially prevalent in this biome. We present data on the distribution and abundance of extrafloral nectaries in the woody flora of cerrado communities and in the flora of other habitats worldwide, and stress the relevance of liquid food sources (including hemipteran honeydew) for the ant fauna. Consumption by ants of plant and insect exudates significantly affects the activity of the associated herbivores of cerrado plant species, with varying impacts on the reproductive output of the plants. Experiments with an ant-plant-butterfly system unequivocally demonstrate that the behavior of both immature and adult lepidopterans is closely related to the use of a risky host plant, where intensive visitation by ants can have a severe impact on caterpillar survival. We discuss recent evidence suggesting that the occurrence of liquid rewards on leaves plays a key role in mediating the foraging ecology of foliage-dwelling ants, and that facultative ant-plant mutualisms are important in structuring the community of canopy arthropods. Ant mediated effects on cerrado herbivore communities can be revealed by experiments performed on wide spatial scales, including many environmental factors such as soil fertility and vegetation structure. We also present some research questions that could be rewarding to investigate in this major neotropical savanna. PMID- 15551027 TI - Queen movement during colony emigration in the facultatively polygynous ant Pachycondyla obscuricornis. AB - In ants, nest relocations are frequent but nevertheless perilous, especially for the reproductive caste. During emigrations, queens are exposed to predation and face the risk of becoming lost. Therefore the optimal strategy should be to move the queen(s) swiftly to a better location, while maintaining maximum worker protection at all times in the new and old nests. The timing of that event is a crucial strategic issue for the colony and may depend on queen number. In monogynous colonies, the queen is vital for colony survival, whereas in polygynous colonies a queen is less essential, if not dispensable. We tested the null hypothesis that queen movement occurs at random within the sequence of emigration events in both monogynous and polygynous colonies of the ponerine ant Pachycondyla obscuricornis. Our study, based on 16 monogynous and 16 polygynous colony emigrations, demonstrates for the first time that regardless of the number of queens per colony, the emigration serial number of a queen occurs in the middle of all emigration events and adult ant emigration events, but not during brood transport events. It therefore appears that the number of workers in both nests plays an essential role in the timing of queen movement. Our results correspond to a robust colony-level strategy since queen emigration is related neither to colony size nor to queen number. Such an optimal strategy is characteristic of ant societies working as highly integrated units and represents a new instance of group-level adaptive behaviors in social insect colonies. PMID- 15551028 TI - Bulk carbon, oxygen, and hydrogen stable isotope composition of recent resins from amber-producing Hymenaea. AB - Resins of Hymenaea, an angiosperm tree genus known to be a copious resin producer and a major source of amber since the Oligo-Miocene, were collected from a wide range of tropical environments from Latin America and Africa, and analyzed for their carbon, hydrogen, and oxygen stable isotope composition. The average value for delta13C in the resins was found to be -27.0+/-1.3 per thousand, which is very similar to the values reported for resins in other studies. Delta18O values for the Hymenaea resins averaged +11.2+/-1.6 per thousand, or about 20 per thousand more depleted than normal plant cellulose. DeltaD values of the resins ranged from -196 to -319 per thousand, with an average of -243+/-30 per thousand. Rough estimates suggest a fractionation of -200 to -210 per thousand between the resins and the environmental water. This value is similar to the -200 per thousand value observed for the fractionation between other plant lipids and environmental water. The present study suggests that the stable isotope composition of fossil resins (amber) has the potential to provide information on ancient environmental waters. PMID- 15551029 TI - Retinal cryptochrome in a migratory passerine bird: a possible transducer for the avian magnetic compass. AB - The currently discussed model of magnetoreception in birds proposes that the direction of the magnetic field is perceived by radical-pair processes in specialized photoreceptors, with cryptochromes suggested as potential candidate molecules mediating magnetic compass information. Behavioral studies have shown that magnetic compass orientation takes place in the eye and requires light from the blue-green part of the spectrum. Cryptochromes are known to absorb in the same spectral range. Because of this we searched for cryptochrome (CRY) in the retina of European robins, Erithacus rubecula, passerine birds that migrate at night. Here, we report three individually expressed cryptochromes, eCRY1a, eCRY1b, and eCRY2. While eCRY1a and eCRY2 are similar to the cryptochromes found in the retina of the domestic chicken, eCRY1b has a unique carboxy (C)-terminal. In light of the 'radical-pair' model, our findings support a potential role of cryptochromes as transducers for the perception of magnetic compass information in birds. PMID- 15551030 TI - [Indian chef with amnesia and muscle pain]. PMID- 15551031 TI - [MRT of the liver. Clinical significance of nonspecific and liver-specific MRT contrast agents]. AB - The liver is a common site for various benign and malignant focal lesions. The initial modality for assessing liver lesions is ultrasound or CT. MRI with its superior soft tissue contrast offers multiple advantages over other imaging modalities. Contrast agents have been developed that increase the detection rate and provide more specific information in comparison to unenhanced techniques. In the mean time three classes are available for MR imaging of the liver: extracellular gadolinium chelates, hepatobiliary and reticulo-endothelia, superparamagnetic agents. We describe in this review the most common focal lesions, their diagnostic possibilities, and the imaging protocols. Clinical use of these contrast agents facilitates detection and differential diagnosis of focal liver lesions that may help to avoid invasive procedures such as biopsy for lesion characterization. PMID- 15551033 TI - Geographical patterns of genetic variation in the world collections of wild annual Cicer characterized by amplified fragment length polymorphisms. AB - Cicer reticulatum, C. echinospermum, C. bijugum, C. judaicum, C. pinnatifidum, C. cuneatum and C. yamashitae are wild annual Cicer species and potential donors of valuable traits to improve chickpea (C. arietinum). As part of a large project to characterize and evaluate wild annual Cicer collections held in the world gene banks, AFLP markers were used to study genetic variation in these species. The main aim of this study was to characterize geographical patterns of genetic variation in wild annual Cicer germplasm. Phylogenetic analysis of 146 wild annual Cicer accessions (including two accessions in the perennial C. anatolicum and six cultivars of chickpea) revealed four distinct groups corresponding well to primary, secondary and tertiary gene pools of chickpea. Some possible misidentified or mislabelled accessions were identified, and ILWC 242 is proposed as a hybrid between C. reticulatum and C. echinospermum. The extent of genetic diversity varied considerably and was unbalanced between species with greatest genetic diversity found in C. judaicum. For the first time geographic patterns of genetic variation in C. reticulatum, C. echinospermum, C. bijugum, C. judaicum and C. pinnatifidum were established using AFLP markers. Based on the current collections the maximum genetic diversity of C. reticulatum, C. echinospermum, C. bijugum and C. pinnatifidum was found in southeastern Turkey, while Palestine was the centre of maximum genetic variation for C. judaicum. This information provides a solid basis for the design of future collections and in situ conservation programs for wild annual Cicer. PMID- 15551034 TI - Molecular markers linked to papaya ring spot virus resistance and Fusarium race 2 resistance in melon. AB - In melon, the Fom-1 gene confers monogenic resistance against the soil-borne fungus Fusarium oxysporum f. sp. melonis, races 0 and 2, while the closely linked Prv gene specifies resistance against the papaya ring spot virus. Markers linked to these resistance (R) genes were identified using two recombinant inbred line populations, derived from crosses between Cucumis melo Vedrantais and C. melo PI 161375, and between C. melo Vedrantais and C. melo PI 414723, respectively. Using bulked segregant analysis, as well as systematic scoring of the mapping populations, we developed two amplified fragment length polymorphism markers, two random amplified polymorphic DNA markers and five restriction fragment length polymorphism (RFLP) markers linked to this locus. Four of the RFLP sequences bear homology to nucleotide-binding site-leucine-rich repeat R genes, indicating the presence of a significant R-gene cluster in this locus. Our study provides the most closely linked markers published so far for these important traits. It also improves the resolution of the whole linkage group IX, which was difficult to order in our previous studies. Two of the markers were converted to cleaved amplified polymorphic sequence markers to facilitate their application in marker assisted selection. Testing these two markers in several melon lines revealed different marker haplotypes in the melon germplasm and supported multiple, independent origin of the Fusarium races 0 and 2 resistance trait. PMID- 15551035 TI - Identification of molecular markers linked to the mildew resistance gene Pl-d in apple. AB - Powdery mildew poses a serious problem for apple growers, and resistance to the disease is a major objective in breeding programmes for cultivar improvement. As selective pressure allows pathogens to overcome previously reliable resistances, there is a need for the introduction of novel resistance genes into new breeding lines. This investigation is concerned with the identification of the first set of molecular markers linked to the gene for mildew resistance, Pl-d, from the accession 'D12'. As no prior information on the map position or markers for Pl-d were available, a bulked-segregant approach was used to test 49 microsatellite primers, 176 amplified fragment length polymorphism (AFLP) primers and 80 random amplified polymorphic DNA (RAPD) primers in a progeny segregating for Pl-d resistance, 'Fiesta' (susceptible) x A871-14 ('Worcester Pearmain' x 'D12'). The segregations of the markers identified in the resistant and susceptible bulks were scored in the progeny, then the recombination fractions between Pl-d and the most tightly linked markers were calculated and a map prepared. Three AFLP, one RAPD and two microsatellite markers were identified. One AFLP was developed into a sequence-characterised amplified region marker, while the microsatellites CH03C02 and CH01D03 were flanking markers, 7 and 11 recombination units, respectively, from Pl-d. Two more distant microsatellites on the same linkage group, CH01D09 and CH01G12, confirmed the orientation of the markers on the linkage group. These microsatellites place Pl-d on the bottom of linkage group 12 in published apple maps, a region where a number of other disease resistance genes have been identified. PMID- 15551036 TI - Comparison of four flow cytometric SNP detection assays and their use in plant improvement. AB - Single nucleotide polymorphisms (SNPs) are attractive DNA markers due to their abundance and potential for use in automated high-throughput genotyping. Numerous SNP genotyping assays have been developed, but it is unclear which assays are best suited and most efficient for various types of plant improvement research. The objective of this study was to compare the accuracy, efficiency, and cost of four SNP genotyping assays: single-base extension (SBE), allele-specific primer extension (ASPE), oligonucleotide ligation (OL), and direct hybridization (DH). All four assay methods used the same Luminex 100 flow cytometer platform. Fifty eight F(2)-derived soybean [Glycine max (L.) Merr.] lines from a cross between inbred lines G99-G725 and N00-3350 were genotyped at four SNPs. SBE and ASPE clearly differentiated between the two homozygotes and the heterozygote at each SNP. Results were in agreement with those identified using the SNaPshot minisequencing assay as a control. In contrast, the OL and DH assays were unable to differentiate between genotypes at some of the SNPs. However, when the cost per data point for the four different assays was compared, the cost of OL and DH was only about 70% of that for SBE, with DH requiring the least time of the four assays. On the basis of cost and labor, ASPE is more cost-effective and simpler than SBE, and would therefore be a good method for genetic mapping and diversity studies which require a large number of markers and a high level of multiplexing. DH appears to be the most economical assay for marker-assisted selection, though optimization for DH would be required for some SNP markers. PMID- 15551037 TI - Two quality-associated HMW glutenin subunits in a somatic hybrid line between Triticum aestivum and Agropyron elongatum. AB - High-molecular-weight glutenin subunits (HMW-GSs) from hybrid line II-12 between wheat (Triticum aestivum L.) and Agropyron elongatum (Host) Nivski were characterized with SDS-PAGE. Out of these HMW-GSs, two subunits, h1Bx and h1By, had mobilities similar to the subunits 1Bx13 and 1By16 from common wheat 4072, which was used as control. Polyclonal antibodies (pAbs) of h1Bx and h1By were prepared, and Western blotting showed that the pAbs had strong affinities for h1Bx and h1By, separately. The specificity of h1Bx-pAb was further checked; it preferentially recognized subunits h1Bx and 1Bx13. HMW-GS gene coding sequences were amplified by genomic polymerase chain reaction from hybrid II-12. Two of the five amplicons, marked II2a and II31b, were sequenced. Their coding sequences are clustered to Glu-1Bx7 and Glu-1By9 of common wheat. Three discrepant regions in deduced amino acid sequences of II2a and 31b repeated one time more than Glu-1Bx7 and Glu-1By9. N-terminal sequences of h1Bx and h1By were determined, which were identical to the published sequences of 1Bx13 and 1By16 and in agreement with that deduced from II2a and II31b, respectively. These results indicated that the two novel genes separated from the hybrid wheat derived from the allelic variation of 1Bx7 and 1By9 of the parent wheat. There is an additional cysteine residue positioned at 271st amino acid of the mature peptide of II2a, which may be related to the high quality of the flour. PMID- 15551038 TI - Use of new EST markers to elucidate the genetic differences in grain protein content between European and North American two-rowed malting barleys. AB - A population comprising 102 doubled haploid lines were produced from a cross between Beka, a barley cultivar widely grown in Spain, and Logan, a north American cultivar with inherently low protein content, a character considered to derive from the cultivar Karl. The intentions were to determine whether low nitrogen malting barleys could be developed in Spain, and if genetic factors that influenced protein content were similarly expressed in widely diverse environments, i.e. northeastern Spain and eastern Scotland. An extensive map comprising 187 molecular markers was developed. Expressed sequence-tagged-derived markers were used in addition to anonymous simple sequence repeats to determine the potential for identifying candidate genes for quantitative trait loci (QTLs), and 22 such markers were mapped for the first time. There was transgressive segregation for both yield and protein content, and the gene for low protein from Logan was not expressed in the Scottish environment. In 2002, high yield was associated with earlier heading date in Spain, while late heading at the Scottish site was associated with greater lodging and lower thousand-kernel weight. These appeared to be possible pleiotropic effects of a factor detected on chromosome 2H. Using information from a consensus map, it was shown that this locus on 2H was in the region of the photoperiod response gene Eam6. A QTL explaining 18% of the variation in grain protein content was detected on chromosome 5H in a region in which a gene for nitrate reductase was previously observed. No effect on grain protein was associated with chromosome 6H, which has been suggested as the location of the low protein gene from Karl. However, it is likely that Karl contained more than one genetic factor reducing protein, and we postulate that the gene on 6H may have been lost during the breeding of Logan. PMID- 15551039 TI - Detection and mapping of QTL for earliness components in a bread wheat recombinant inbred lines population. AB - Earliness, an adaptative trait and factor of variation for agronomic characters, is a major trait in plant breeding. Its constituent traits, photoperiod sensitivity (PS), vernalization requirement (VR) and intrinsic earliness (IE), are largely under independent genetic controls. Mapping of major genes and quantitative trait loci (QTL) controlling these components is in progress. Most of the studies focusing on earliness considered it as a whole or through one (or two) of its components. The purpose of this study was to detect and map QTL for the three traits together through an experimental design combining field trials and controlled growth conditions. QTL were mapped in a population of F(7) recombinant inbred lines derived by single-seed descent from a cross between two French varieties, 'Renan' and 'Recital'. A map was previously constructed, based on 194 lines and 254 markers, covering about 77% of the genome. Globally, 13 QTL with a LOD>2.5 were detected, of which four control PS, five control VR and four control IE. Two major photoperiod sensitive QTL, together explaining more than 31% of the phenotypic variation, were mapped on chromosomes 2B and 2D, at the same position as the two major genes Ppd-B1 and Ppd-D1. One major VR QTL explaining (depending on the year) 21.8-39.6% of the phenotypic variation was mapped on 5A. Among the other QTL, two QTL of PS and VR not referenced so far were detected on 5A and 6D, respectively. A VR QTL already detected on 2B in a connected population was confirmed. PMID- 15551040 TI - Use of trial clustering to study QTL x environment effects for grain yield and related traits in maize. AB - A population of 300 F(3:4) lines derived from the cross between maize inbred lines F2 and F252 was evaluated for testcross value in a large range of environmental conditions (11 different locations in 2 years: 1995 and 1996) in order to study (1) the magnitude of genotype x environment and (2) the stability of quantitative trait loci (QTL) effects. Several agronomic traits were measured: dry grain yield (DGY), kernel weight, average number of kernels per plant, silking date (SD) and grain moisture at harvest. A large genotype x environment interaction was found, particularly for DGY. A hierarchical classification of trials and an additive main effects and multiplicative interaction (AMMI) model were carried out. Both methods led to the conclusion that trials could be partitioned into three groups consistent with (1) the year of experiment and (2) the water availability (irrigated vs non-irrigated) for the trials sown in 1995. QTL detection was carried out for all the traits in the different groups of trials. Between 9 and 15 QTL were detected for each trait. QTL x group and QTL x trial effects were tested and proved significant for a large proportion of QTL. QTL detection was also performed on coordinates on the first two principal components (PC) of the AMMI model. PC QTL were generally detected in areas where QTL x group and QTL x trial interactions were significant. A region located on chromosome 8 near an SD QTL seemed to play a key role in DGY stability. Our results confirm the key role of water availability and flowering earliness on grain yield stability in maize. PMID- 15551041 TI - Selection of international molecular standards for DNA fingerprinting of Theobroma cacao. AB - A collaborative international program was initiated to identify and describe the genetic diversity of living germplasm collections of Theobroma cacao genotypes that are maintained in several international collections scattered throughout tropical cacao growing countries of the world. Simple sequence repeat (SSR) DNA analysis was identified as the most appropriate molecular tool for DNA fingerprinting these collections during an international forum representing academic, government and industry scientists in the cacao community. Twenty-five SSR primers, which had been previously described, were evaluated as potential candidates to define an efficient, standardized, molecular fingerprinting protocol for T. cacao accessions. These primers have been evaluated for reliability, widespread distribution across the cacao genome, number of alleles produced by the SSR primers in cacao and their ability to discriminate between cacao accessions. Approximately 690 cacao accessions were used to evaluate the utility of these SSR primers as international molecular standards, and a small number of test samples of T. cacao were sent to two other independent laboratories for verification. DNA fragments were selectively amplified by PCR, using the SSR primers labeled with fluorescent dyes, and separated by capillary electrophoresis. Based on this study, the 15 SSR primers that had the highest reproducibility and consistency within a common genotype, while allowing the differentiation of separate divergent genotypes, were selected as international molecular standards for DNA fingerprinting of T. cacao. PMID- 15551042 TI - Identification of quantitative trait loci controlling resistance to maize chlorotic dwarf virus. AB - Ineffective screening methods and low levels of disease resistance have hampered genetic analysis of maize (Zea mays L.) resistance to disease caused by maize chlorotic dwarf virus (MCDV). Progeny from a cross between the highly resistant maize inbred line Oh1VI and the susceptible inbred line Va35 were evaluated for MCDV symptoms after multiple virus inoculations, using the viral vector Graminella nigrifrons. Symptom severity scores from three rating dates were used to calculate area under the disease progress curve (AUDPC) scores for vein banding, leaf twist and tear, and whorl chlorosis. AUDPC scores for the F(2) population indicated that MCDV resistance was quantitatively inherited. Genotypic and phenotypic analyses of 314 F(2) individuals were compared using composite interval mapping (CIM) and analysis of variance. CIM identified two major quantitative trait loci (QTL) on chromosomes 3 and 10 and two minor QTL on chromosomes 4 and 6. Resistance was additive, with alleles from Oh1VI at the loci on chromosomes 3 and 10 contributing equally to resistance. PMID- 15551043 TI - QTL detection for eating quality of cooked rice in a population of chromosome segment substitution lines. AB - The genetic mechanism underlying six palatability properties of cooked rice and three physico-chemical traits was dissected in 66 BC(3)F(2) chromosome segment substitution lines (CSSLs), using a complete linkage map in three successive years. The CSSLs showed transgressive segregation for all traits studied. Significant correlation was detected among most palatability traits. A total of 25 QTLs for the nine traits were identified on nine chromosomes, and many QTLs affecting different quality traits were mapped in the same regions. Six QTLs--qLT 8 for luster, qTD-6 and qTD-8 for tenderness, qIVOE-6 and qIVOE-8 for integrated value of organoleptic evaluation, and qAC-8 for amylose content--were repeatedly detected across the 3 years. Phenotypic values were significantly different between the recurrent parent, cultivar Asominori, and the CSSLs harboring any of the six QTL alleles across the three environments, indicating that these six QTLs were non-environment-specific and could be used for marker-assisted selection in rice quality improvement. PMID- 15551045 TI - Prolonged cardiac repolarisation during spontaneous nocturnal hypoglycaemia in children and adolescents with type 1 diabetes. AB - AIMS/HYPOTHESIS: It has been postulated that hypoglycaemia-related cardiac dysrhythmia and, in particular, prolonged cardiac repolarisation, may contribute to increased mortality rates in children and adolescents with type 1 diabetes. METHODS: We examined the prevalence of prolonged QT interval on ECG during spontaneous hypoglycaemia in 44 type 1 diabetic subjects (aged 7-18 years), and explored the relationships between serial overnight measurements of QT interval corrected for heart rate (QTc) and serum glucose, potassium and epinephrine levels. Each subject underwent two overnight profiles; blood was sampled every 15 min for glucose measurements and hourly for potassium and epinephrine. Serial ECGs recorded half-hourly between 23.00 and 07.00 hours were available on 74 nights: 29 with spontaneous hypoglycaemia (defined as blood glucose <3.5 mmol/l) and 45 without hypoglycaemia. RESULTS: Mean overnight QTc was longer in females than in males (412 vs 400 ms, p=0.02), but was not related to age, diabetes duration or HbA(1)c. Prolonged QTc (>440 ms) occurred on 20 out of 74 (27%) nights, with no significant differences between male and female subjects, and was more prevalent on nights with hypoglycaemia (13/29, 44%) than on nights without (7/45, 15%, p=0.0008). Potassium levels were lower on nights when hypoglycaemia occurred (minimum potassium 3.4 vs 3.7 mmol/l, p=0.0003) and were inversely correlated with maximum QTc (r=-0.40, p=0.03). In contrast, epinephrine levels were not higher on nights with hypoglycaemia and were not related to QTc. CONCLUSIONS/INTERPRETATION: In young type 1 diabetic subjects, prolonged QTc occurred frequently with spontaneous overnight hypoglycaemia and may be related to insulin-induced hypokalaemia. PMID- 15551046 TI - Short-term infusion of interleukin-6 does not induce insulin resistance in vivo or impair insulin signalling in rats. AB - AIMS/HYPOTHESIS: Interleukin-6 has been implicated in the insulin resistance associated with obesity and impaired glucose tolerance. Previous studies in vitro have shown that IL-6 rapidly (1-2 h) impairs cellular insulin signalling and action through an increased expression of suppressor of cytokine signalling (SOCS)-3. In the present study, IL-6 or saline was infused in rats that were simultaneously in a state of hyperinsulinaemia. Muscle, liver and adipose tissue were excised after 2 h to examine potential effects on insulin signalling or gene expression. METHODS: The rats were infused with IL-6 or saline during a euglycaemic-hyperinsulinaemic clamp and the glucose infusion rate was measured after 90 to 120 min. Signal transducer and activator of transcription (STAT)3 phosphorylation and insulin-stimulated tyrosine phosphorylation of the insulin receptors and IRS were measured with immunoblotting and gene expression through real-time PCR. RESULTS: No inhibitory effect of IL-6 on insulin-stimulated whole body glucose uptake was seen in spite of high circulating levels of IL-6 (0.85+/ 0.08 nmol/l). Tyrosine phosphorylation of the insulin receptors and IRS was also unchanged in the liver, skeletal muscles and adipose tissue. However, tyrosine phosphorylation of STAT3 was increased in all tissues, showing that IL-6 signalling was activated. IL-6 mRNA tended to increase, while GLUT4, peroxisome proliferator-activated receptor-gamma coactivator-1 (PGC-1) and adiponectin gene expression were unchanged. CONCLUSIONS/INTERPRETATION: Infusion of IL-6 for 120 min in rats during euglycaemic-hyperinsulinaemic conditions did not alter the effect of insulin on whole-body glucose homeostasis, plasma adiponectin levels or insulin signalling in target tissues. Thus, the acute effects of IL-6, associated with SOCS-3 induction, do not lead to whole-body insulin resistance. These data further underscore the importance of the chronic, and potentially tissue-specific effects of IL-6 on insulin signalling and action. PMID- 15551047 TI - Aldosterone escape during blockade of the renin-angiotensin-aldosterone system in diabetic nephropathy is associated with enhanced decline in glomerular filtration rate. AB - AIMS/HYPOTHESIS: It has been suggested that aldosterone plays a role in the initiation and progression of renal disease independently of arterial blood pressure and plasma angiotensin II levels. We evaluated the influence of plasma aldosterone levels on progression of diabetic nephropathy during long-term blockade of the renin-angiotensin-aldosterone system. METHODS: A total of 63 hypertensive patients with type 1 diabetes and diabetic nephropathy were treated with losartan, 100 mg once daily, for a mean follow-up period of 35 months. Plasma aldosterone, GFR, albuminuria and 24-h blood pressure were determined at baseline and at regular intervals during the study. RESULTS: Patients were divided according to their increasing or decreasing levels of plasma aldosterone during long-term losartan treatment in an escape group (n=26) and a non-escape group (n=37). In the escape group, aldosterone levels increased from (geometric mean [95% CI]) 57 pg/ml (43-76 pg/ml) at 2 months, to 102 pg/ml (78-134 pg/ml) at the end of the study (p<0.01). The corresponding levels in the non-escape group were 83 pg/ml (69-102 pg/ml) and 49 pg/ml (40-60 pg/ml; p<0.01). The median rate of decline in GFR was 5.0 ml.min(-1).year(-1) (range 0.4-15.9 ml.min(-1).year( 1)) in the escape group, compared with 2.4 ml.min(-1).year(-1) (-1.6 to 11.0 ml.min(-1).year(-1)) in the non-escape group (p<0.005). The increase in plasma aldosterone correlated with the rate of decline in GFR (r(2)=0.19, p<0.001), corresponding to a decline in GFR of 1.5 ml.min(-1).year(-1) for every two-fold increase in plasma aldosterone. Pre-treatment and treatment values of plasma aldosterone were not related to albuminuria or to changes in albuminuria during the study. CONCLUSIONS/INTERPRETATION: Our data suggest that aldosterone escape during long-term blockade of the renin-angiotensin-aldosterone system is associated with an enhanced decline in GFR in patients with type 1 diabetes and diabetic nephropathy. PMID- 15551048 TI - Visualising insulin secretion. The Minkowski Lecture 2004. AB - Insulin secretion from pancreatic islet beta cells is a tightly regulated process, under the close control of blood glucose concentrations, neural inputs and circulating hormones. Defects in glucose-triggered insulin secretion, possibly exacerbated by a decrease in beta cell mass, are ultimately responsible for the development of type 2 diabetes. A full understanding of the mechanisms by which glucose and other nutrients trigger insulin secretion will probably be essential to allow for the development of new therapies of type 2 diabetes and for the derivation of "artificial" beta cells from embryonic stem cells as a treatment for type 1 diabetes. I focus here on recent developments in our understanding of beta cell glucose sensing, achieved in part through the development of recombinant targeted probes (luciferase, green fluorescent protein) that allow islet beta cell metabolism and Ca(2+) handling to be imaged in situ in the intact islet with single cell resolution. Combined with classical biochemistry, these techniques show that the beta cell is uniquely poised, thanks to the expression of low levels of lactate dehydrogenase and plasma membrane lactate/monocarboxylate transporters, to channel glucose carbons towards oxidative metabolism, ATP synthesis and inhibition of AMP-activated protein kinase, a newly defined regulator of insulin release. I also discuss the molecular basis of the recruitment of secretory vesicles to the cell surface, analysed by the use of new imaging techniques including total internal reflection of fluorescence, as well as the "nanomechanics" of the exocytotic event itself. PMID- 15551049 TI - [The infected implant. Part 1: bacteriology]. AB - Periprosthetic infection is a significant complication in joint replacement surgery and develops in 0.5-2% of cases. Staphylococcus aureus and commensal microorganisms of the skin, especially coagulase-negative staphylococci, as well as a broad spectrum of other potential pathogens typically already colonize the surface of the foreign body at the time of implantation. Specific mechanisms such as bacterial adhesion to host factors absorbed in the material, biofilm formation, and a metabolic adaptation of adherent microorganisms play a particularly important role in the pathogenesis and course of the disease. Microbiological diagnosis requires to some extent complex culture procedures of puncture specimens or tissue removed during surgery; this can be supplemented by modern molecular testing. Antimicrobial treatment must be conceived as a synopsis of clinical picture, confirmed pathogen, and the intended surgical procedure on an individual basis and is routinely administered as combination therapy for several weeks, sometimes also as sequential therapy. Validated preventive measures in joint replacement surgery include mandatory perioperative antibiotic prophylaxis and other additional measures. PMID- 15551050 TI - [Tissue engineering of bone. Integration and migration of human mesenchymal stem cells in colonized contructs in a murine model]. AB - Tissue engineering opens up new ways for therapy of bone defects. Therefore, the aim of this study was to establish a mouse model to investigate local cell growth of human mesenchymal stem cells (hMSC) on the scaffold in vivo. Moreover, migration of cells to other organs should be excluded.hMSC (Cambrex, USA) were cultivated according to supplier's recommendations. After inoculation on cylindric scaffolds, one matrix cell construct and one scaffold without hMSC were implanted subcutaneously left and right paravertebrally in athymic nude mice. After 2, 4, 8, and 12 weeks constructs and organs were harvested for immunohistological evaluation and PCR. In conclusion, we found integration of scaffolds loaded with hMSC implanted ectopically. HMSC seeded on 3D scaffolds survived for a period of up to 12 weeks. In addition, we could not detect hMSC in any other organ of the host. PMID- 15551051 TI - Loss of endothelium-dependent relaxation in abdominal aorta preserved in a co storage system. AB - The potentially detrimental influence of parenchymal cells on endothelial function during preservation in UW solution was examined by co-storage of rat abdominal aortic rings with isolated liver cells. Cold storage of rings in UW solution alone for up to 96 h had no effect on the response to acetylcholine, though constriction was progressively lost. Co-storage of rings with liver cells resulted in no loss of sodium nitroprusside response, but the relaxation response to acetylcholine was reduced. The loss of acetylcholine response could not be attributed to Kupffer cells, the lowering of pH, oxygen depletion, or the loss of constriction. A similar loss of endothelial function was observed in rings stored in pieces of liver, kidney or heart. We conclude that parenchymal cells exude factors during preservation by cold storage which reversibly inhibit vascular NO production. These factors could significantly impair whole organ function on reperfusion. PMID- 15551052 TI - In vitro assessment of a new ABO immunosorbent with synthetic carbohydrates attached to sepharose. AB - Transplantation across the ABO barrier is sometimes done in cases of emergency, such as acute liver failure, but is also carried out in elective cases, e.g. kidneys from living donors. Reducing the recipient anti-A/B antibody titres is often necessary in ABO-incompatible kidney transplantation. This is usually done by the use of techniques such as plasmapheresis and protein A- or sepharose linked anti-human Ig immunoadsorption. A new ABO immunosorbent with synthetic A- or B-trisaccharide carbohydrate epitopes linked to a sepharose matrix has been tested. Columns made of this material have been tested in vitro with plasma from A- and B-individuals, assessed for antibody reduction capacity, flow characteristics, biocompatibility, and unspecific protein adsorption. The columns have a high capacity for ABO antibody removal, reducing titres by three to seven steps in one passage. We noted a high biocompatibility, with no unspecific protein adsorption, no activation of coagulation factors, and a low activation of complement, no immune complex formation and no cytotoxicity towards cultured mammalian L929 cells. PMID- 15551055 TI - Decreased bone mineral density and content in neurofibromatosis type 1: lowest local values are located in the load-carrying parts of the body. AB - Neurofibromatosis type 1 (NF1) is a dominantly inherited disease. Skeletal ailments such as short stature, kyphoscoliosis, tibial bowing and pseudarthrosis are common osseous manifestations of NF1. Previously, a correlation with scoliosis and decreased bone mineral density (BMD) of the lumbar spine has been reported in 12 NF1 patients. A total of 35 NF1 patients and 26 healthy controls were included in the present study. Of the participants over 20 years of age (26 NF1 patients and all controls) 14 were male and 12 were female, seven of whom were premenopausal. The controls were matched for age, sex and body mass index (BMI). Physical activity and medical history of NF1 patients were evaluated to screen the fractures and osseous manifestations of the disease and to rule out the factors that effect BMD. BMD and bone mineral content (BMC) were measured with DXA, using a total body program. The present study detected a lowered bone mineral density (p =0.028) and content (p <0.001) in NF1 patients of both sexes. The results of the present study also show that NF1 patients have an increased risk for osteoporosis. Among NF1 patients seven cases of osteoporosis and 13 cases of osteopenia were detected. In controls, one case of osteoporosis and 13 cases of osteopenia were detected. The location of the lowest local BMD was clustered to the load-carrying parts of the body in NF1 patients. Physical activity and the medical history of the NF1 patients did not explain the decreased BMD and BMC. The findings of the present and previous studies suggest that the pathogenesis of the osseous manifestations in NF1 may involve impaired development of the skeletal system and impaired maintenance of bone structure. PMID- 15551056 TI - Bone remodeling: new aspects of a key process that controls skeletal maintenance and repair. AB - Bone remodeling is the concerted interplay of two cellular activities: osteoclastic bone resorption and osteoblastic bone formation. Bone remodeling is the physiologic process that maintains bone mass, skeletal integrity and skeletal function. A molecular understanding of this process is therefore of paramount importance for almost all aspects of skeletal physiology and many facets of bone diseases. Based on the morphological observation of the BMU-"bone multicellular unit" or "bone metabolic unit"-and a wide body of in vitro data, bone remodeling was thought to be controlled locally through functional coupling of resorption and formation and vice versa. However, recent genetic studies have shown that there is no obligatory tight cross-control of bone formation and bone resorption in vivo and that there is also a central axis controlling bone formation, one aspect of bone remodeling. The molecule that inhibits bone formation through a hypothalamic relay is leptin. Following binding to its receptor located on the ventromedial nuclei of the hypothalamus, leptin's action on bone formation is mediated via a neuronal signaling cascade that involves the ss-adrenergic system. The overall goal of this review is to show how the dialogue between clinical medicine and mouse genetics helped to uncover a new concept in skeletal physiology. PMID- 15551057 TI - Additive effects of estrogen and mechanical stress on nitric oxide and prostaglandin E2 production by bone cells from osteoporotic donors. AB - Mechanical loading is thought to provoke a cellular response via loading-induced flow of interstitial fluid through the lacuno-canalicular network of osteocytes. This response supposedly leads to an adaptation of local bone mass and architecture. It has been suggested that loss of estrogen during menopause alters the sensitivity of bone tissue to mechanical load, thereby contributing to the rapid loss of bone. The present study aimed to determine whether estrogen modulates the mechanoresponsiveness of bone cells from osteoporotic women. Bone cell cultures from nine osteoporotic women (aged 62-90 years) were pre-cultured for 24 h with 10(-11) mol/l 17beta-estradiol (E2) or vehicle, and subjected to 1 h of pulsating fluid flow (PFF) or static culture. E2 alone enhanced prostaglandin E(2) (PGE(2)) and nitric oxide (NO) production by 2.8-fold and 2.0 fold, respectively, and stimulated endothelial nitric oxide synthase protein expression by 2.5-fold. PFF, in the absence of E2, stimulated PGE(2) production by 3.1-fold and NO production by 3.9-fold. Combined treatment with E2 and PFF increased PGE(2) and NO production in an additive manner. When expressed as PFF treatment-over-control ratio, the response to fluid shear stress was similar in the absence or presence of E2. These results suggest that E2 does not affect the early response to stress in bone cells. Rather, E2 and shear stress both promote the production of paracrine factors such as NO and PGE(2) in an additive manner. PMID- 15551058 TI - Vitamin D status as the major factor determining the circulating levels of parathyroid hormone: a study in normal subjects. AB - We investigated the relative contribution of the major factors regulating calcium homeostasis in determining the circulating levels of PTH. We studied 137 males and 125 females who were healthy volunteers. Circulating PTH levels were determined by three different immunoradiometric assays (IRMA). The first one (PTH Sorin, PTH S) utilizes two affinity-purified polyclonal antibodies directed against the 1-34 and 39-84 sequence of the hormone. The two other IRMA share polyclonal anti-PTH (39-84) antibodies. The first assay (PTH Whole, PTH W) utilizes a second polyclonal antibody, directed against the 1-4 amino acid sequence. The second assay (PTH Total, PTH T) utilizes a second antibody specific for the 7-34 region. Concentrations of PTH fragments lacking the initial amino acid sequence (PTH N-truncated, PTH N-t) were determined by the difference of values between PTH T and PTH W. Vitamin D was the main explicative variable almost in every multiple linear regression model, both considering the group as a whole (PTH S: R2 = 0.238, P < 0.0001; PTH W: R2 = 0.08, P < 0.001; PTH T: R2 = 0.145, P < 0.0001; PTH N-t: R2 = 0.081, P < 0.009) and when considering men and women separately. In subjects with vitamin D insufficiency (n = 53) [25(OH)D < 30 nmol/l], mean serum levels of parathyroid hormone were significantly higher (P < 0.001) than those in subjects of similar age with normal vitamin status (n = 209) with all the assays employed. This study demonstrates the central role of 25(OH)D in regulating PTH secretion in physiological conditions. PMID- 15551059 TI - The genome sequence of an anaerobic aromatic-degrading denitrifying bacterium, strain EbN1. AB - Recent research on microbial degradation of aromatic and other refractory compounds in anoxic waters and soils has revealed that nitrate-reducing bacteria belonging to the Betaproteobacteria contribute substantially to this process. Here we present the first complete genome of a metabolically versatile representative, strain EbN1, which metabolizes various aromatic compounds, including hydrocarbons. A circular chromosome (4.3 Mb) and two plasmids (0.21 and 0.22 Mb) encode 4603 predicted proteins. Ten anaerobic and four aerobic aromatic degradation pathways were recognized, with the encoding genes mostly forming clusters. The presence of paralogous gene clusters (e.g., for anaerobic phenylacetate oxidation), high sequence similarities to orthologs from other strains (e.g., for anaerobic phenol metabolism) and frequent mobile genetic elements (e.g., more than 200 genes for transposases) suggest high genome plasticity and extensive lateral gene transfer during metabolic evolution of strain EbN1. Metabolic versatility is also reflected by the presence of multiple respiratory complexes. A large number of regulators, including more than 30 two component and several FNR-type regulators, indicate a finely tuned regulatory network able to respond to the fluctuating availability of organic substrates and electron acceptors in the environment. The absence of genes required for nitrogen fixation and specific interaction with plants separates strain EbN1 ecophysiologically from the closely related nitrogen-fixing plant symbionts of the Azoarcus cluster. Supplementary material on sequence and annotation are provided at the Web page http://www.micro-genomes.mpg.de/ebn1/. PMID- 15551062 TI - Cutaneous metabolism of glycol ethers. AB - The toxicity of glycol ethers is associated with their oxidation to the corresponding aldehyde and alkoxyacetic acid by cytosolic alcohol dehydrogenase (ADH; EC 1.1.1.1.) and aldehyde dehydrogenase (ALDH; 1.2.1.3). Dermal exposure to these compounds can result in localised or systemic toxicity including skin sensitisation and irritancy, reproductive, developmental and haemotological effects. It has previously been shown that skin has the capacity for local metabolism of applied chemicals. Therefore, there is a requirement to consider metabolism during dermal absorption of these compounds in risk assessment for humans. Cytosolic fractions were prepared from rat liver, and whole and dermatomed skin by differential centrifugation. Rat skin cytosolic fractions were also prepared following multiple dermal exposure to dexamethasone, ethanol or 2 butoxyethanol (2-BE). The rate of ethanol, 2-ethoxyethanol (2-EE), ethylene glycol, 2-phenoxyethanol (2-PE) and 2-BE conversion to alkoxyacetic acid by ADH/ALDH in these fractions was continuously monitored by UV spectrophotometry via the conversion of NAD+ to NADH at 340 nm. Rates of ADH oxidation by rat liver cytosol were greatest for ethanol followed by 2-EE >ethylene glycol >2-PE >2-BE. However, the order of metabolism changed to 2-BE >2-PE >ethylene glycol >2-EE >ethanol using whole and dermatomed rat skin cytosolic fractions, with approximately twice the specific activity in dermatomed skin cytosol relative to whole rat skin. This suggests that ADH and ALDH are localised in the epidermis that constitutes more of the protein in dermatomed skin than whole skin cytosol. Inhibition of ADH oxidation in rat liver cytosol by pyrazole was greatest for ethanol followed by 2-EE >ethylene glycol >2-PE >2-BE, but it only inhibited ethanol metabolism by 40% in skin cytosol. Disulfiram completely inhibited alcohol and glycol ether metabolism in the liver and skin cytosolic fractions. Although ADH1, ADH2 and ADH3 are expressed at the protein level in rat liver, only ADH1 and ADH2 are selectively inhibited by pyrazole and they constitute the predominant isoforms that metabolise short-chain alcohols in preference to intermediate chain-length alcohols. However, ADH1, ADH3 and ADH4 predominate in rat skin, demonstrate different sensitivities to pyrazole, and are responsible for metabolising glycol ethers. ALDH1 is the predominant isoform in rat liver and skin cytosolic fractions that is selectively inhibited by disulfiram and responds to the amount of aldehyde formed by the ADH isoforms expressed in these tissues. Thus, the different affinity of ADH and ALDH for alcohols and glycol ethers of different carbon-chain length may reflect the relative isoform expression in rat liver and skin. Following multiple topical exposure, ethanol metabolism increased the most following ethanol treatment, and 2-BE metabolism increased the most following 2-BE treatment. Ethanol and 2-BE may induce specific ADH and ALDH isoforms that preferentially metabolise short-chain alcohols (i.e. ADH1, ALDH1) and longer chain alcohols (i.e. ADH3, ADH4, ALDH1), respectively. Treatment with a general inducing agent such as dexamethasone enhanced ethanol and 2-BE metabolism suggesting induction of multiple ADH isoforms. PMID- 15551063 TI - Changes in expression of bcl-2 and bax in Syrian hamster embryo (SHE) cells exposed to ZnCl2. AB - Zinc is involved in many physiological processes and plays a critical role in functional and structural cells. Zinc at concentrations ranging from 100 to 150 micromol L(-1) has been shown to induce morphological transformation of Syrian hamster embryo (SHE) cells. At these concentrations, zinc inhibited apoptosis in SHE cells. The objective of this study was to elucidate the mechanisms of action of zinc on the apoptotic pathway. Effects of 100 and 150 micromol L(-1) ZnCl(2) on the expression of two members of the Bcl-2 family of proteins and on the transcription factor c-Myc in SHE cells was investigated using RT-PCR. No effect on the proto-oncogene c-myc was observed. Up-regulation of bcl-2 expression was found and bax expression was reduced. These changes have been corroborated by immunoblotting. Effects of Zn(2+) on bcl-2/bax ratio were confirmed in apoptotic camptothecin-treated SHE cells. Cloned and sequenced cDNAs obtained from RT-PCR amplifications allowed us to check the RT-PCR products encoded the expected proteins. This study demonstrated that zinc acts in the early phases of the apoptotic process by modification of the bcl-2/bax ratio in normal and apoptotic SHE cells. PMID- 15551065 TI - Angiotensin II blocks memory consolidation through an AT2 receptor-dependent mechanism. AB - RATIONALE AND OBJECTIVES: Several studies suggest that the brain renin angiotensin system is involved in memory consolidation. However, the participation of angiotensin II (AII) in this process is controversial. This is probably due to the fact that many of the studies carried out to elucidate this matter employed multitrial learning paradigms together with pretraining intracerebroventricular infusions, and therefore were unable to distinguish between consolidation and retrieval related events and lacked anatomical specificity. To circumvent this problem, we analyzed the role played in memory consolidation by AII using the hippocampal-dependent, one-trial, step-down inhibitory avoidance task (IA) in combination with stereotaxically localized intrahippocampal infusion of drugs. METHODS AND RESULTS: Rats bilaterally implanted with infusion cannulae into the CA1 region of the dorsal hippocampus (CA1) were trained in IA and tested for memory retention 24 h later. We found that when infused into CA1 immediately or 30 min after training but not later, AII produced a dose-dependent amnesic effect without altering locomotor activity, exploratory behavior or anxiety state. The amnesic effect of AII was not mimicked by angiotensin IV (AIV) and was totally blocked by the AII-type 2 receptor (AT2) antagonist, PD123319, but not by the AII-type 1 receptor (AT1) antagonist, losartan. Importantly, when infused alone, neither PD123319 nor losartan produced any effect on memory retention. CONCLUSIONS: Our data indicate that, when given into CA1, AII blocks memory formation through a mechanism involving activation of AT2 receptors; however, endogenous AII does not seem to participate in the consolidation of IA long-term memory. PMID- 15551068 TI - Role of alpha-2 adrenoceptors in stress-induced reinstatement of alcohol seeking and alcohol self-administration in rats. AB - RATIONALE AND OBJECTIVES: Alpha-2 adrenoceptors are known to be involved in stress-induced reinstatement of heroin and cocaine seeking in laboratory animals. Here, we studied the involvement of these receptors in stress-induced reinstatement of alcohol seeking by using an agonist (lofexidine) and an antagonist (yohimbine) of these receptors, which inhibit and activate, respectively, noradrenaline transmission. We also tested the effect of lofexidine and yohimbine on alcohol self-administration. Lofexidine is used clinically for treating opiate withdrawal symptoms and yohimbine induces stress-like responses in humans and non-humans. METHODS: Rats were trained to self-administer alcohol (12% w/v, 1 h/day) and after extinction of the alcohol-reinforced behavior, they were tested for the effect of lofexidine (0, 0.05 and 0.1 mg/kg, IP) on reinstatement of alcohol seeking induced by intermittent footshock stress (10 min, 0.8 mA) or for the effect of yohimbine (0, 1.25 and 2.5 mg/kg, IP) on reinstatement of alcohol seeking. Other rats were trained to self-administer alcohol, and after stable responding, the effects of lofexidine and yohimbine on alcohol self-administration were determined. RESULTS: Pretreatment with lofexidine (0.05 mg/kg and 0.1 mg/kg) attenuated stress-induced reinstatement of alcohol seeking and also decreased alcohol self-administration. In contrast, yohimbine pretreatment potently reinstated alcohol seeking after extinction and also induced a profound increase in alcohol self-administration. CONCLUSIONS: Results indicate that activation of alpha-2 adrencoceptors is involved in both stress-induced reinstatement of alcohol seeking and alcohol self-administration. To the degree that the present results are relevant to human alcoholism, alpha-2 adrencoceptor agonists should be considered in the treatment of alcohol dependence. PMID- 15551071 TI - Microanalytical flow-through method for assessment of the bioavailability of toxic metals in environmental samples. AB - The application of a recently proposed microanalytical flow-through system for on line sequential extraction of heavy metals from solid samples of environmental interest is described. Using various extraction schemes (a nitric acid scheme, a two-stage extraction scheme using two reagents applied in the BCR procedure) and comparison with the common batch sequential BCR procedure, the suitability of the system for fast screening of solid environmental samples is demonstrated. By pumping leaching agents sequentially through the sample held in a micro cartridge, the different metal fractions present can be assessed in less than an hour. Method evaluation was performed using SRM 1648 urban particulate matter and BCR 701 lake sediment reference material certified for extractable metals. The need for and design of laboratory internal reference material suitable for simulating the natural (dynamic) processes of metal release into the environment is also discussed. For the first time correlation is sought between fractionation techniques and physiologically based methods for assessment of the bioaccessibility of metals in biomatrices. PMID- 15551069 TI - Differential effects of intra-midbrain raphe and systemic 8-OH-DPAT on VTA self stimulation thresholds in rats. AB - RATIONALE: Intra-median raphe nucleus (MRN) administration of the 5-HT(1A) receptor agonist 8-OH-DPAT decreases lateral hypothalamic self-stimulation thresholds and is reported to have biphasic effects following systemic administration. These experiments attempted to extend the previous findings to mesolimbic pathway self-stimulation at ventral tegmental area (VTA) electrodes. OBJECTIVES: This study was conducted to provide comparative data for systemic and intra-dorsal raphe nucleus (DRN) and intra-MRN effects of 8-OH-DPAT on VTA self stimulation. METHODS: Male Sprague-Dawley rats with VTA electrodes were trained to respond for electrical stimulation. Systemic and intra-midbrain raphe 8-OH DPAT effects on rate-frequency thresholds were measured. Systemic administration of WAY 100635 was used to confirm 5-HT(1A) receptor mediation of 8-OH-DPAT effects. RESULTS: 8-OH-DPAT (0.003-0.3 mg kg(-1) SC) increased rate-frequency thresholds and decreased maximal response rates. WAY 100635 alone (0.0125-0.1 mg kg(-1) SC) did not alter these measures. Intra-DRN and intra-MRN 8-OH-DPAT (5.0 microg) decreased rate-frequency thresholds without altering maximal response rates. Intra-DRN 8-OH-DPAT (0.1-5.0 microg) induced a slight decrease and intra MRN 8-OH-DPAT a slight increase in locomotor activity. WAY 100635 (0.1 mg kg(-1)) blocked effects of 8-OH-DPAT on VTA self-stimulation. CONCLUSION: These results confirm threshold-decreasing effects of intra-MRN 8-OH-DPAT and extend this to the DRN and to VTA thresholds. Monophasic dose dependent increases in VTA thresholds following systemic 8-OH-DPAT are not equivalent to reports for hypothalamic self-stimulation. Differences between studies may be attributable to stimulation site and/or differences in threshold measurement procedures. Effects of WAY 100635 in this study indicate 5-HT(1A) receptor mediation of these 8-OH DPAT effects. PMID- 15551072 TI - New dimension in nano-imaging: breaking through the diffraction limit with scanning near-field optical microscopy. AB - In recent years scanning near-field optical microscopy (SNOM) has developed into a powerful surface analytical technique for observing specimens with lateral resolution equal to or even better than 100 nm. A large number of applications, from material science to biology, have been reported. In this paper, two different kinds of near-field optical microscopy, aperture and scattering-type SNOM, are reviewed together with recent studies in surface analysis and biology. Here, near-field optical techniques are discussed in comparison with related methods, such as scanning probe and standard optical microscopy, with respect to their specific advantages and fields of application. PMID- 15551073 TI - An assay for determination of rat adrenal catechol-O-methyltransferase activity: comparison of spontaneously hypertensive rats and Wistar-Kyoto rats. AB - A method has been developed for measurement of catechol-O-methyltransferase (COMT) activity in the rat adrenal gland. Epinephrine, synthesized in the adrenal gland, was used as substrate, and its enzymatic product, metanephrine, was quantified by high-performance liquid chromatography (HPLC) with fluorescence detection. The method has sufficient precision and accuracy. Soluble (S) and membrane-bound (MB) COMT activity in Wistar-Kyoto (WKY) rats was 20.7 +/- 3.5 and 18.6 +/- 3.4 pmol min(-1) mg(-1) protein (n = 5), respectively. To clarify the role of adrenal COMT in blood-pressure regulation, S and MB COMT activity in spontaneously hypertensive rats were determined. Respective activity was 18.6 +/- 3.4 and 17.0 +/- 1.1 pmol min(-1) mg(-1) (n = 5), which is similar to that in WKY rats. This finding suggests that COMT in the adrenal gland might not be related to blood pressure regulation. PMID- 15551074 TI - Detection of proteins cross-linked within galactoside polyacrylate-based hydrogels by means of a quantum dot fluororeagent. AB - Protein toxins have been immobilized in a galactoside polyacrylate hydrogel in a microarray format. The large pore size and solution-like environment of these novel hydrogels allow for easy penetration of large proteins and detection reagents. Confocal microscopy provided three-dimensional visualization of dye labeled toxins cross-linked within the gel and of streptavidin-coated quantum dot (QD) fluorophores used to visualize the toxins after incubation with biotinylated anti-toxin antibodies. Fluorescence microscopy was utilized to visualize arrays of toxins detected by a biotinylated antibody and then exposure to streptavidin conjugated QDs. The intensity of the QD fluorescence was quantified, and binding to two toxins on three types of hydrogels was examined. PMID- 15551075 TI - Measurement of water by oven evaporation using a novel oven design. 2. Water in motor oils and motor oil additives. AB - The measurement of water in lubricating oils is important because water accelerates the corrosion of metal parts and bearings in motors. Some of the additives added to lubricating oils to improve their performance react with the Karl Fischer reagent (KFR) causing a positive bias in the water measurement. A new oven evaporation technique for measuring water in oils has been developed that is automated, requires less sample handling, is easily calibrated, and is capable of measuring relatively small mass fractions of water (> or =50 mg/kg sample). A series of motor oils was analyzed with the standard KFR, a reagent that detects interfering substances that reduce iodine, and the aldehyde-ketone reagent that does not detect substances that react with methanol and form water. The oil samples were heated to 107 degrees C and then reheated to 160 degrees C. At both temperatures, material was measured by both KFRs, but only zinc dithiophosphate released sulfur compounds that would react with the reagent that detects interfering substances. Mass fractions of between 20 and 70% of the volatile material released at either temperature were measured with the standard KFR but not with the aldehyde-ketone reagent. These results demonstrate that there are a number of sources of positive bias in the measurement of water in motor oils and that the standard KFR cannot be used to measure water in motor oils and motor oil additives. These results also indicate that some of the material reacts with methanol to form water. Finally, these results suggest that some of the material that is volatile at 160 degrees C and not at 107 degrees C may be water that is physically occluded or may be substances that react with diethyleneglycol monomethylether to produce water. PMID- 15551076 TI - Depth profile studies of ZrTiN coatings by laser ablation inductively coupled plasma mass spectrometry. AB - The feasibility of depth profiling was studied by using a 193-nm ArF* excimer laser ablation system (GeoLas, MicroLas, Goettingen, Germany) with a lens array based beam homogenizer in combination with an ICP-QMS Agilent 7500. Two ablation cells (20 and 1.5 cm3) were compared at the laser repetition rate of 1 Hz, laser beam energy of 135 mJ and the carrier gas flow rate 1.5 L min(-1) He + 0.78 L min(-1) Ar. The ablation cell dimensions are important parameters for signal tailing; however, very small cell volumes (e.g. 1.5 cm3) may cause memory effects, which can be probably explained by dominant inertial losses of aerosol on cell walls with its delayed mobilization. The 20-cm3 ablation cell seems to be appropriate for depth profiling by continuous single-hole drilling. The study of the influence of the pit diameter magnitude on the waning and emerging signals under small crater depth/diameter aspect ratios, which range between 0.75 and 0.0375 for the 3-microm-thick coatings and pit diameters 4-80 microm, revealed that the steady-state signals of pure coating and pure substrate (out of interface) were obtained at crater diameters between 20 and 40 microm. Depth resolution defined by means of slopes of tangents in the layer interface region depend on the pit diameter and has an optimum value between 20 and 40 microm and gives 0.6 microm for the 20-microm pit. In-depth variation of concentration of coating constituent (Ti) was proved to be almost identical with two different laser/ICP systems. PMID- 15551077 TI - Simultaneous multichannel mass-specific detection for high-performance liquid chromatography using an array detector sector-field mass spectrometer. AB - The use of a separation step, such as liquid chromatography, prior to inductively coupled plasma mass spectrometry (ICP-MS) has become a common tool for highly selective and sensitive analyses. This type of coupling has several benefits including the ability to perform speciation analysis or to remove isobaric interferences. Several limitations of conventional instruments result from the necessity to scan or pulse the mass spectrometer to obtain a complete mass spectrum. When the instrument is operated in such a non-continuous manner, duty cycle is reduced, resulting in poorer absolute limits of detection. Additionally, with scanning instruments, spectral skew can be introduced into the measurement, limiting quantitation accuracy. To address these shortcomings, a high-performance liquid chromatograph has been coupled to an ICP-MS capable of continuous sample introduction and simultaneous multimass detection. These features have been realized with a novel detector array, the focal plane camera. Instrument performance has been tested for both speciation analysis and for the elimination of isobaric interferences. Absolute limits of detection in the sub picogram to tens of picograms regime are obtainable, while the added mass dimension introduced by simultaneous detection dramatically increases chromatographic peak capacity. PMID- 15551078 TI - Torsional eye movements during psychophysical testing with rotating patterns. AB - Torsional eye movements were measured while subjects viewed a large, high contrast windmill pattern rotating at 53 degrees /s or a small (5 degrees diameter) dot pattern rotating at 115 degrees /s. Both stimuli generated rotational eye movements consisting of torsional optokinetic nystagmus (tOKN) superimposed on a slow torsional drift in the direction of pattern rotation. With the wide-field windmill stimulus, torsional drifts of up to 7 degrees over 20 s were found. The dot pattern produced drifts of up to 2 degrees over 5-20 s. In both cases, the slow-phase speeds during tOKN were low (0.5-1 degrees /s). We conclude that reductions in slip speed are minimal with rotating stimuli, so torsional eye speeds will have a minimal effect on investigations of rotational motion aftereffect strength and perceived speed. While the slow-phase tOKN gain is low, the slow drift in torsional eye position will have significant effects on psychophysical results when the tests rely on keeping selected regions of the stimulus confined to specific areas of the retina, as is the case for phantom or remote motion aftereffects. PMID- 15551079 TI - Effects of neonatal C-fiber depletion on the integration of paired-whisker inputs in rat barrel cortex. AB - In the present study we used computer-controlled mechanical displacement of paired whiskers in normal and C-fiber-depleted rats to quantitatively examine the role of C-fibers in the receptive field properties of barrel cortical cells. In rodents when adjacent whiskers are stimulated prior to the main whisker responses to the main whisker are inhibited, the degree of inhibition being a function of the inter-deflection intervals. The adjacent-whisker-evoked inhibition of barrel cells in normal and C-fiber-depleted rats using neonatal capsaicin treatment were examined by stimulation of the adjacent whisker zero, 10, 20, 30, 50 and 100 ms prior to the main whisker deflection. C-fiber depletion reduced the suppressive effect of paired whisker stimulation at all of the tested inter-stimulus intervals without changing response latencies. The main effect was observed during the later phase of response (about 13-17 ms from stimulus onset) and not during the initial responses (7-12 ms). These results suggest that the inhibitory receptive field properties of low-threshold mechanical somatosensory cells are influenced by C-fibers. PMID- 15551080 TI - What the hand can't tell the eye: illusion of space constancy during accurate pointing. AB - When we press an elevator button or pick up a coffee cup, different visual information is used to guide our reach and to form our conscious experience of these objects. But can the information guiding our hand be brought into awareness? The fact that we can see and feel our own hand in action suggests that it might be possible. However, the dual visual systems theory claims that on-line control of movement is governed by the dorsal stream of visual processing, which is largely unconscious. Two experiments are presented as strong tests of the hypothesis that the visual information guiding on-line pointing in healthy human adults is inaccessible for conscious report. Results show that participants are incapable of consciously accessing the information used in pointing, even though they can see and feel their hands in action and accurate performance depends on it. PMID- 15551081 TI - Behavioral plasticity of antisaccade performance following daily practice. AB - The ability to change behavior to adapt to the environment, known as behavioral plasticity, is an important part of daily life. In the present study subjects' performances on antisaccade tasks were manipulated by training them on one of three different eye movement tasks (antisaccade, prosaccade, and fixation). Thirty subjects were tested at three time points over a 2-week period and practiced their assigned task every day between test sessions. Subjects who trained on antisaccades significantly decreased their error rates, while maintaining their reaction time, suggesting that accuracy did not improve at the expense of speed. Subjects who practiced the prosaccade task made more errors on the antisaccade task on subsequent test sessions, while those who practiced the fixation task showed no change across test sessions. These results suggest that deliberate practice of eye movement tasks can alter antisaccade performance, and that the direction of the effect is dependent upon the type of practice in which the subject engages. PMID- 15551082 TI - Effects of sympathetic stimulation on the rhythmical jaw movements produced by electrical stimulation of the cortical masticatory areas of rabbits. AB - The somatomotor and sympathetic nervous systems are intimately linked. One example is the influence of peripheral sympathetic fibers on the discharge characteristics of muscle spindles. Since muscle spindles play important roles in various motor behaviors, including rhythmic movements, the working hypothesis of this research was that changes in sympathetic outflow to muscle spindles can change rhythmic movement patterns. We tested this hypothesis in the masticatory system of rabbits. Rhythmic jaw movements and EMG activity induced by long lasting electrical cortical stimulation were powerfully modulated by electrical stimulation of the peripheral stump of the cervical sympathetic nerve (CSN). This modulation manifested itself as a consistent and marked reduction in the excursion of the mandibular movements (often preceded by a transient modest enhancement), which could be attributed mainly to corresponding changes in masseter muscle activity. These changes outlasted the duration of CSN stimulation. In some of the cortically evoked rhythmic jaw movements (CRJMs) changes in masticatory frequency were also observed. When the jaw-closing muscles were subjected to repetitive ramp-and-hold force pulses, the CRMJs changed characteristics. Masseter EMG activity was strongly enhanced and digastric EMG slightly decreased. This change was considerably depressed during CSN stimulation. These effects of CSN stimulation are similar in sign and time course to the depression exerted by sympathetic activity on the jaw-closing muscle spindle discharge. It is suggested that the change in proprioceptive information induced by an increase in sympathetic outflow (a) has important implications even under normal conditions for the control of motor function in states of high sympathetic activity, and (b) is one of the mechanisms responsible for motor impairment under certain pathological conditions such as chronic musculoskeletal head-neck disorders, associated with stress conditions. PMID- 15551083 TI - Long-lasting modulation of human motor cortex following prolonged transcutaneous electrical nerve stimulation (TENS) of forearm muscles: evidence of reciprocal inhibition and facilitation. AB - Several lines of evidence indicate that motor cortex excitability can be modulated by manipulation of afferent inputs, like peripheral electrical stimulation. Most studies in humans mainly dealt with the effects of prolonged low-frequency peripheral nerve stimulation on motor cortical excitability, despite its being known from animal studies that high-frequency stimulation can also result in changes of the cortical excitability. To investigate the possible effects of high-frequency peripheral stimulation on motor cortical excitability we recorded motor-evoked potentials (MEPs) to transcranial magnetic stimulation (TMS) of the left motor cortex from the right flexor carpi radialis (FCR), extensor carpi radialis (ECR), and first dorsal interosseous (FDI) in normal subjects, before and after transcutaneous electrical nerve stimulation (TENS) of 30 min duration applied over the FCR. The amplitude of MEPs from the FRC was significantly reduced from 10 to 35 min after TENS while the amplitude of MEPs from ECR was increased. No effects were observed in the FDI muscle. Indices of peripheral nerve (M-wave) and spinal cord excitability (H waves) did not change throughout the experiment. Electrical stimulation of the lateral antebrachial cutaneous nerve has no significant effect on motor cortex excitability. These findings suggest that TENS of forearm muscles can induce transient reciprocal inhibitory and facilitatory changes in corticomotoneuronal excitability of forearm flexor and extensor muscles lasting several minutes. These changes probably may occur at cortical site and seem to be mainly dependent on stimulation of muscle afferents. These findings might eventually lead to practical applications in rehabilitation, especially in those syndromes in which the excitatory and inhibitory balance between agonist and antagonist is severely impaired, such as spasticity and dystonia. PMID- 15551084 TI - The effects of practice and delay on motor skill learning and retention. AB - The present study assessed the effects of amount of practice and length of delay on the learning and retention of a timed motor sequence task. Participants learned to reproduce ten-element visual sequences by tapping in synchrony with the stimulus. Participants were randomly assigned to a varied-practice condition or a varied-delay condition. In the varied-practice condition, participants received either one, three, or six blocks of practice followed by a fixed 4-week delayed-recall. In the varied-delay condition, participants received three blocks of practice followed by a varied delay of either 3 days, or 2, 4, or 8 weeks. Learning was assessed by changes in accuracy, response variance, and percent response asynchrony. Our results showed that amount of practice per se did not affect learning and retention of the task. Rather, distribution of practice over several days was the most important factor affecting learning and retention. We hypothesize that passage of time is essential for a maximum benefit of practice to be gained, as the time delay may allow for consolidation of learning, possibly reflecting plastic changes in motor cortical representations of the skill. With regards to delay, our findings suggest that explicit and motoric components of a motor sequence are likely to be learned and maintained in separate but interacting systems. First, only the longest delay group showed decrements in percent correct, indicating that longer lengths of delay might hinder retrieval of explicit aspects of the task. Second, all groups showed a decrement in percent response asynchrony, suggesting that synchronization may be a more difficult parameter to maintain because it relies heavily on sensorimotor integration. PMID- 15551085 TI - Task dependent gain regulation of spinal circuits projecting to the human flexor carpi radialis. AB - In humans, the flexor carpi radialis (FCR) and extensor carpi radialis (ECR) muscles act as antagonists during wrist flexion-extension and as functional synergists during radial deviation. In contrast to the situation in most antagonist muscle pairs, Renshaw cells innervated by the motor neurons of each muscle inhibit the motoneurons, but not Ia inhibitory interneurons, of the opposite motor pool. Here we compared gain regulation of spinal circuits projecting to FCR motoneurons during two tasks: flexion and radial deviation of the wrist. We also investigated the functional consequences of this organisation for maximal voluntary contractions (MVCs). Electromyographic (EMG) recordings were taken from FCR, ECR longus and ECR brevis using fine-wire electrodes and electrical stimulation was delivered to the median and radial nerves. Ten volunteers participated in three experiments. 1. To study the regulation of the Renshaw cell-mediated, inhibitory pathway from ECR to FCR motoneurons, forty stimuli were delivered to the radial nerve at 50% of the maximal M-wave amplitude for ECR brevis. Stimuli were delivered during both isometric wrist flexions and radial deviation actions with an equivalent EMG amplitude in FCR (approximately 5% wrist flexion MVC). 2. To explore the homonymous Ia afferent pathway to FCR motoneurons, 50 stimuli were delivered to the median nerve at intensities ranging from below motor threshold to at least two times that which evoked a maximal M wave during wrist flexion and radial deviation (matched FCR EMG at approximately 5% wrist flexion MVC). 3. EMG amplitude was measured during MVCs in wrist flexion, extension and radial deviation. There was no significant difference in the inhibition of FCR EMG induced via ECR-coupled Renshaw cells between radial deviation and wrist flexion. However, the mean FCR H-reflex amplitude was significantly (P<0.05) greater during wrist flexion than radial deviation. Furthermore, EMG amplitude in FCR and ECR brevis was significantly (P<0.05) greater during MVCs in wrist flexion and extension (respectively) than radial deviation. ECR longus EMG was significantly greater during MVCs in radial deviation than extension. These results indicate that the gain of the Renshaw mediated inhibitory pathway between ECR and FCR motoneurons is similar for weak flexion and radial deviation actions. However, the gain of the H-reflex pathway to FCR is greater during wrist flexion than radial deviation. Transmission through both of these pathways probably contributes to the inability of individuals to maximally activate FCR during radial deviation MVCs. PMID- 15551086 TI - Perception of non-voluntary brief contractions in normal subjects and in a deafferented patient. AB - The accuracy of force perception by human subjects in the absence of voluntary motor command was evaluated by exploring how they perceived isometric twitches of wrist extensor muscles produced by external stimulation. Twelve normal subjects and a well-known patient lacking large-diameter afferent fibres (GL) performed estimation, production and reproduction tasks. Magnetic stimulation of the radial nerve and, in normal subjects, transcranial magnetic stimulation (TMS) of the motor cortex were used to produce weak brief non-voluntary twitches. In estimation tasks, the subjects had to ascribe verbal marks on a 1-5 scale to the forces of stimulation-induced twitches. Loose covariations of marks and forces were observed, while directions of force variations between successive twitches were relatively well detected. GL did less well than normal subjects in detecting directions of force variations. In production tasks, subjects had to produce twitches matching verbal command marks in a 1-5 range, with or without visual feedback. Performances of normal subjects and GL resembled those of estimation tasks and were not improved by visual feedback. In reproduction tasks, subjects had to duplicate stimulation-induced test twitches: first without visual feedback, second with and third again without. Large errors were observed but all subjects did better with visual feedback. In the third step, improvement with respect to the first one was significantly more marked with TMS than with peripheral stimulation. GL improved her performance in the third step, possibly because she could use information provided by group III and group IV afferents still present in her nerves. Altogether, for normal subjects (1) the performances in estimation tasks are consistent with the known behaviour of Golgi tendon organs as observed in animal experiments, and (2) results observed in reproduction tasks suggest that cortical stimulation might elicit, in addition to corticospinal activation of motoneurones, collateral discharges that could be stored as a memory of motor command. PMID- 15551087 TI - Interlimb transfer of load compensation during rapid elbow joint movements. AB - Previous research has shown that training of a novel task can improve subsequent performance in the opposite arm owing to anticipation of the previously learned task conditions. Interestingly, we recently reported preliminary evidence that such transfer might also include modulation of feedback-mediated responses. We now test interlimb transfer of load compensation responses, measured through kinematic and EMG recordings during rapid 20 degrees elbow flexion movements. Two subject groups, LR and RL, each comprising six right-handed subjects, first performed using either the left (LR) or right (RL) arm, followed by opposite arm performance. After 30 trials of consistent performance, five random trials within a background of 50 trials were loaded with a 2-kg mass prior to the "go" signal. We compared load compensation responses for naive performance with those following opposite arm exposure. Under naive conditions, the resulting load compensation responses began about 50 ms following movement onset, and were substantially more effective for the nondominant arm. Opposite arm exposure substantially improved the accuracy of only dominant arm responses. This, however, did not occur through changes in the short latency components of the load compensation response. Instead, changes in muscle activities, associated with interlimb transfer, began some 150 ms following movement onset. We expect that these changes represent transfer in the "volitional" component of the load compensation response. Because the shorter latency response was unaffected by opposite arm exposure, modulation of this component likely requires prior experience with limb specific effectors. PMID- 15551088 TI - EMG, force and discharge rate analysis of human jaw reflexes in response to axial stimulation of the incisor. AB - Reflex studies utilising controlled stimulation along the long axis of human incisors are relatively new, and the effects that various stimulus parameters have on the elicited reflexes are not fully understood. Twelve subjects were recruited to determine the effects that contraction level, stimulus force and amount of constant force applied between stimuli have on the reflex response of the masseter muscle. Multi-unit intramuscular electromyogram (EMG) was recorded alongside surface EMG to determine whether any differences existed between the two. Furthermore, cumulative peri-stimulus "dischargegrams" were constructed to determine whether events seen in the EMG corresponded to changes in the discharge rates of the underlying motor units. Axial stimulation of the incisor induced a response in the EMG comprising of peak-trough-peak, with the trough being the most dominant. The bite force record showed only a reduction (relaxation) in response to the stimulation. The most significant experimental factor affecting the reflex occurrence and strength was the stimulus force. Although the latency, duration and occurrence rates were not significantly different, the strength of the responses was greater in intramuscular recordings compared with the surface recordings. Discharge rate analysis showed that approximately two-thirds of the late peaks detected in the EMG did not correspond to an increase in the discharge rates of the underlying units; hence they were due to the clustering of action potentials following the trough and not to a change in the membrane potential of the motoneurone. It was also found that the duration of the trough, as seen by the reduced cumulative discharge rate of the underlying units, was longer than indicated by the EMG. PMID- 15551089 TI - Pitch discrimination accuracy in musicians vs nonmusicians: an event-related potential and behavioral study. AB - Previously, professional violin players were found to automatically discriminate tiny pitch changes, not discriminable by nonmusicians. The present study addressed the pitch processing accuracy in musicians with expertise in playing a wide selection of instruments (e.g., piano; wind and string instruments). Of specific interest was whether also musicians with such divergent backgrounds have facilitated accuracy in automatic and/or attentive levels of auditory processing. Thirteen professional musicians and 13 nonmusicians were presented with frequent standard sounds and rare deviant sounds (0.8, 2, or 4% higher in frequency). Auditory event-related potentials evoked by these sounds were recorded while first the subjects read a self-chosen book and second they indicated behaviorally the detection of sounds with deviant frequency. Musicians detected the pitch changes faster and more accurately than nonmusicians. The N2b and P3 responses recorded during attentive listening had larger amplitude in musicians than in nonmusicians. Interestingly, the superiority in pitch discrimination accuracy in musicians over nonmusicians was observed not only with the 0.8% but also with the 2% frequency changes. Moreover, also nonmusicians detected quite reliably the smallest pitch changes of 0.8%. However, the mismatch negativity (MMN) and P3a recorded during a reading condition did not differentiate musicians and nonmusicians. These results suggest that musical expertise may exert its effects merely at attentive levels of processing and not necessarily already at the preattentive levels. PMID- 15551090 TI - Predictive strategies in interception tasks: differences between eye and hand movements. AB - To investigate how the sensorimotor systems of eye and hand use position, velocity, and timing information of moving targets, we conducted a series of three experiments. Subjects performed combined eye-hand catch-up movements toward visual targets that moved with step-ramp-like velocity profiles. Visual feedback of the hand was prevented by blanking the target at the onset of the hand movement. A multiple regression was used to determine the effects of position, velocity, and timing accessed before each movement on the movement amplitudes of eye and hand. The following results were obtained: 1. The predictive strategy of eye movements could be modeled by a linear regression on the basis of the position error and the target velocity. This was not the case for hand movements, for which there was a significant partial correlation between the movement amplitude and the product of target velocity and movement duration. This correlation was not observed for eye movements suggesting that the predictive strategy of hand movements takes movement duration into account, in contrast to the strategy used in eye movements. 2. To determine whether the movement amplitudes of eye and hand depend on a categorical classification between a discrete number of movement types, we compared an experiment in which target position and velocity were distributed continuously with an experiment using only four different combinations of target position and velocity. No systematic differences between these experiments were observed. This shows that the system output is a function of continuous, interval-scaled variables rather than a function of discrete categorical variables. 3. We also analyzed the component of the movement amplitudes not explained by the regression, i.e., the residual error. The residual errors between subsequent trials were correlated more strongly for eye than for hand movements, suggesting that short-term temporal fluctuations of the predictive strategy were stronger for the eye than for the hand. PMID- 15551091 TI - "Acoustical vision" of below threshold stimuli: interaction among spatially converging audiovisual inputs. AB - Crossmodal spatial integration between auditory and visual stimuli is a common phenomenon in space perception. The principles underlying such integration have been outlined by neurophysiological and behavioral studies in animals; this study investigated whether the integrative effects observed in animals also apply to humans. In this experiment we systematically varied the spatial disparity (0 degrees, 16 degrees, and 32 degrees) and the temporal interval (0, 100, 200, 300, 400, and 500 ms) between the visual and the auditory stimuli. Normal subjects were required to detect visual stimuli presented below threshold either in unimodal visual conditions or in crossmodal audiovisual conditions. Signal detection measures were used. An enhancement of the perceptual sensitivity (d') for luminance detection was found when the audiovisual stimuli followed a simple spatial and temporal rule, governing multisensory integration at the neuronal level. PMID- 15551092 TI - Locally induced hypothermia for treatment of acute ischaemic stroke: a physical feasibility study. AB - During the treatment of stroke by local intra-arterial thrombolysis (LIT) it is frequently possible to pass the blood clot with a micro-catheter, allowing perfusion of brain tissue distally to the occlusion. This possibility allows for new early treatments of ischaemic brain tissue, even before the blood clot has been removed. One potential new approach to preserve brain tissue at risk may be locally induced endovascular hypothermia. Physical parameters such as the required micro-catheter input pressure, output velocity and flow rates, and a heat exchange model, applicable in the case of a micro-catheter placed within a guiding catheter, are presented. Also, a simple cerebral temperature model is derived that models the temperature response of the brain to the perfusion with coolant fluids. Based on this model, an expression has been derived for the time needed to reach a certain cerebral target temperature. Experimental in vitro measurements are presented that confirm the usability of standard commercially available micro-catheters to induce local hypothermia of the brain. If applied in vivo, the model predicts a local cooling rate of ischaemic brain tissue of 300 g of approximately 1 degrees C in 1 min, which is up to a factor 30-times faster than the time-consuming systemic hypothermia via the skin. Systemic body temperature is only minimally affected by application of local hypothermia, thus avoiding many limitations and complications known in systemic hypothermia. PMID- 15551093 TI - Pituitary gland signal in pre-term infants during the first year of life: an MRI study. AB - Our purpose was to determine the pituitary gland signal of premature neonates and infants. We retrospectively measured, in a midline sagittal T1-weighted MRI scan, the pituitary signal of 121 premature infants (mean gestational age: 32.8 weeks, mean chronological age: 6.9 months, mean corrected age: 5.2 months). The relative signal intensity of the adenohypophysis and neurohypophysis was measured as (A V)/V and (P-V)/V, respectively (where A, P, V are the mean signal intensities of the adenohypophysis, neurohypophysis and vermis white matter, respectively). On visual inspection the adenohypophysis was rated relative to the vermis white matter (VWM) and scored as hyperintense (grade 3), isointense (grade 2) and hypointense (grade 1). (A-V)/V and (P-V)/V had a negative correlation with chronological age (r=-0.38, r=-0.35, respectively, P<0.01) and corrected age (r= 0.42, r=-0.40, respectively, P<0.01). The (A-V)/V was different in the three grade groups; significant difference was found between grades 3 and 2 and grades 3 and 1. The adenohypophysis was bright in 93% of babies under 2 months of chronological age. In five babies of mean gestational age 29.1 weeks a hyperintense adenohypophysis was observed at 2.9-3.7 months of chronological age (corrected age 0.37-2 months). We conclude that in pre-term babies the adenohypophysis appears in hypersignal that may persist up to 2 months of corrected age. The adenohypophyseal and neurohypophyseal signals decrease with age. PMID- 15551094 TI - Basal cell adenoma of prostate. AB - A case of basal cell adenoma of the prostate accompanying benign prostatic hypertrophy is presented. This is an uncommon, benign lesion which is usually mistaken for a carcinoma. In order to emphasize the importance of differential diagnose and to better define this entity, we present a case report. PMID- 15551096 TI - Acquired combined factor X and XII deficiency with isolated bleeding complications in a patient with AL amyloidosis. PMID- 15551097 TI - Pericentric chromosome 8 inversion associated with the 5'RUNX1/3'CBFA2T1 gene in acute myeloid leukemia cases. AB - In the present paper we report pericentric chromosome 8 inversions in two (2.4%) of 82 acute myeloid leukemia (AML) cases characterized by the 5'RUNX1/3'CBFA2T1 fusion gene. Molecular cytogenetic characterization was achieved using appropriate bacterial artificial chromosome (BAC) and P1 artificial chromosome (PAC) probes in fluorescence in situ hybridization (FISH) experiments. In these two cases the fusion gene was detected on the der(8) short arm, resulting from a pericentric chromosome 8 inversion followed by a t(8;21) rearrangement. These results suggest that heterogeneous mechanisms can lead to the generation of the 5'RUNX1/3'CBFA2T1chimeric gene. PMID- 15551095 TI - Molecular pathways in bladder cancer. AB - The aim of this review is to provide a contemporary outline of our current understanding of the molecular and genetic events associated with tumorigenesis and the progression of bladder cancer. A comprehensive review of the literature was performed on the molecular alterations associated with transitional cell carcinoma (TCC) of the bladder. Intense research efforts are being made to better identify and characterize various bladder cancers and their true biologic potential. The need to predict which superficial tumors will recur or progress, and which invasive tumors will metastasize has led to a much better understanding of the molecular pathways associated with bladder cancer. The molecular changes that occur in TCC of the bladder are numerous and can be categorized into: (1) chromosomal alterations leading to carcinogenesis, (2) loss of cell cycle regulation accounting for cellular proliferation, and (3) metastasis, guided by events such as angiogenesis. It is becoming apparent that the accumulation of genetic and molecular changes ultimately determines a tumors phenotype and subsequent clinical behavior. At the present time, conventional histopathologic evaluation of bladder cancer (tumor grade and stage) is inadequate to accurately predict the behavior of most bladder tumors. While new laboratory techniques have allowed us to better understand how bladder cancer develops and ultimately progresses, few of these techniques are currently available for use in the clinical setting. The ultimate goal is to develop reliable prognostic markers which will accurately predict not only the expected clinical course of an individual bladder tumor but also the response of that tumor to currently available therapies. More importantly, this information may be employed in the future to dictate altogether new treatments for the prevention and/or stabilization of the early molecular events that lead to the development of bladder cancer. PMID- 15551098 TI - Characterization of red cell autoantibodies in consecutive DAT-positive patients with relation to in vivo haemolysis. AB - The clinical symptoms of patients with autoimmune haemolytic anaemia (AIHA) may range from no symptoms at all to severe haemolysis with life-threatening anaemia. The aim of the present study was to characterize the autoantibodies serologically and to evaluate their activity in vitro in monocyte monolayer assay (MMA) in consecutive direct antiglobulin test (DAT)-positive patients from a haematological department. We also aimed to evaluate the monocyte production of cytokines in vitro and relate all findings to in vivo haemolysis. Twenty-nine patients with positive DAT were included in the study and clinical characteristics were documented. The patients were divided into three groups based on laboratory parameters, severe haemolysis, moderate haemolysis and no sign of haemolysis. Severe haemolysis was related to a strongly positive DAT and positive results in MMA. When eluates were analysed in MMA, a positive result was associated with a higher concentration in the supernatants of interleukin (IL)-8 after 6 h incubation. We conclude that MMA activity and in vitro cytokine production may reflect the in vivo activity of autoantibodies. This may be of importance in understanding the mechanisms of haemolysis and in predicting the harmfulness of antibodies, especially when blood transfusions are needed. PMID- 15551099 TI - Electrodynamic smooth muscle sphincter: development and biomechanical evaluation of a novel porcine artificial smooth muscle sphincter in a new in vitro stoma simulator. AB - AIM: Many authors have suggested that the activity of the enteric inhibitory nerves is important in regulating normal gastrointestinal motility and inducing smooth muscle relaxation. Hitherto, no experimental or clinical models exist that transfer these physiological aspects to creating an autologous artificial sphincter for the treatment of major incontinence. Therefore, this study was performed to determine the contractile and relaxant capacity of gastrointestinal muscle types and to investigate the efficiency of a novel smooth muscle sphincter, based on the non-adrenergic, non-cholinergic (NANC) receptive relaxation under electrical field stimulation (EFS). METHODS: For the first step, the isometric tension from isolated circular porcine fundus and colon muscle strips was recorded during pharmacological stimulation (TTX, L-NNA and atropine) and EFS. As a result, a continent electrodynamic smooth muscle sphincter (ESMS) was created by wrapping a fundus muscle flap around an isolated segment of porcine distal colon. The EFS of the free nerve fibers of the flap was realized using a circular platinum wire electrode. Parameters such as threshold of continence, intra/preluminal pressure and fluid passage were analyzed in a newly designed in vitro stoma simulator. RESULTS: Electrical field stimulation produced a maximal and voltage-dependent fundus relaxation to --12.4 mN/mm(2) (frequency of 40 Hz, pulse duration, train duration and voltage of 5 ms, 1 s and 60 mA respectively), which were abolished by N-nitro-L -arginine (L-NNA; 10(-4) M) in a dose-dependent manner, confirming that relaxant responses were mediated by NANC nerves. The results of eight ESMS showed that circular electrical stimulation of the muscle flap caused muscle relaxation with a concomitant and effective reduction in the occlusion pressure. CONCLUSION: The NANC-induced relaxation mechanism of porcine fundus preparations could be transferred to an efficient smooth muscle sphincter with a high threshold of continence and electrically controlled defecation. PMID- 15551100 TI - Comparison of inpatient and outpatient upper gastrointestinal haemorrhage. AB - BACKGROUND AND AIMS: Inpatients developing upper gastrointestinal (GI) haemorrhage are at increased risk of death. This study was performed to elucidate differences in inpatients and outpatients. PATIENTS/METHODS: Three hundred and sixty-two patients who needed esophagogastroduodenoscopy for upper GI bleeding were identified from endoscopy charts. Patients' characteristics, bleeding parameters, clinical presentation, pre-existing medication, and laboratory data were compared between patients who were admitted because of upper GI bleeding and patients who developed bleeding while in hospital for other reasons. RESULTS/FINDINGS: Hospital mortality was 39.0% in inpatients vs. 11.1% in outpatients (p<0.01). Death due to bleeding was observed in 9.5% of inpatients vs. 2.5% of outpatients (p<0.01). Whereas peptic ulcer was the most common source of bleeding in both, variceal bleeding was the most common cause of death because of haemorrhage in both. Recurrent bleeding was associated with mortality in outpatients (p<0.001), but not in inpatients (p=0.11). Rates of bleeding recurrence and need for surgery was similar in both groups. Inpatients suffered more often from renal disease, pulmonary disease, diabetes mellitus, coagulopathy, or immunosuppression, and were treated more frequently with acetylsalicylic acid, glucocorticoids and heparin. The frequency of pre-existing disease was higher in inpatients. INTERPRETATION/CONCLUSION: Higher mortality after GI bleeding in inpatients than in outpatients is due to a generally higher prevalence of co-morbidity rather than a single or a few risk factors. PMID- 15551102 TI - Miniaturized heme proteins: crystal structure of Co(III)-mimochrome IV. AB - Protein design provides an attractive approach to test the essential features required for folding and function. Previously, we described the design and structural characterization in solution of mimochromes, a series of miniaturized metalloproteins, patterned after the F-helix of the hemoglobin beta-chain. Mimochromes consist of two medium-sized helical peptides, covalently linked to the deuteroporphyrin. CD and NMR characterization of the prototype, mimochrome I, revealed that the overall structure conforms well to the design. However, formation of Delta and Lambda diastereomers was observed. To overcome the problem of diastereomer formation, we re-designed mimochrome I, by engineering intramolecular, interchain interactions. The resulting model was mimochrome IV: the solution structural characterization showed the presence of the Lambda isomer as a unique form. To examine the extent to which the stereochemical stability and uniqueness of mimochrome IV was retained in the solid state, the crystal structure of Co(III)-mimochrome IV was solved by X-ray diffraction, and compared to the solution structure of the same derivative. Co(III)-mimochrome IV structures, both in solution and in the solid state, are characterized by the following common features: a bis-His axial coordination, a Lambda configuration around the metal ion, and a predominant helical conformation of the peptide chains. However, in the crystal structure, intrachain Glu1-Arg9 ion pairs are preferred over the designed, and experimentally found in solution, interchain interactions. This ion pairing switch may be related to strong packing interactions. PMID- 15551103 TI - Toward clinically relevant standardization of image quality. AB - In recent years, notable progress has been made on standardization of medical image presentations in the definition and implementation of the Digital Imaging and Communications in Medicine (DICOM) Grayscale Standard Display Function (GSDF). In parallel, the American Association of Physicists in Medicine (AAPM) Task Group 18 has provided much needed guidelines and tools for visual and quantitative assessment of medical display quality. In spite of these advances, however, there are still notable gaps in the effectiveness of DICOM GSDF to assure consistent and high-quality display of medical images. In additions the degree of correlation between display technical data and diagnostic usability and performance of displays remains unclear. This article proposes three specific steps that DICOM, AAPM, and ACR may collectively take to bridge the gap between technical performance and clinical use: (1) DICOM does not provide means and acceptance criteria to evaluate the conformance of a display device to GSDF or to address other image quality characteristics. DICOM can expand beyond luminance response, extending the measurable, quantifiable elements of TG18 such as reflection and resolution. (2) In a large picture archiving and communication system (PACS) installation, it is critical to continually track the appropriate use and performance of multiple display devices. DICOM may help with this task by adding a Device Service Class to the standard to provide for communication and control of image quality parameters between applications and devices, (3) The question of clinical significance of image quality metrics has rarely been addressed by prior efforts. In cooperation with AAPM, the American College of Radiology (ACR), and the Society for Computer Applications in Radiology (SCAR), DICOM may help to initiate research that will determine the clinical consequence of variations in image quality metrics (eg, GSDF conformance) and to define what constitutes image quality from a diagnostic perspective. Implementation of these three initiatives may further the reach and impact of DICOM toward quality medicine. PMID- 15551104 TI - Applying DICOM to dentistry. AB - There are more than 160,000 dentists licensed in the United States. For the dental patient, the dentist is both radiologist and treating clinician. The American Dental Association (ADA) has been a member of the Digital Imaging and Communication in Medicine (DICOM) Standard Committee since 1996. DICOM v.3 provides image object definitions for digital transmission radiography (Dx) with special categorization for intraoral projections (Io), and it also provides for color photography used in dentistry. Digital dental radiographs include transmission images of the head and jaws, pantomography, tomography and cone-beam computed tomography. In 2000, the ADA resolved to strive for interoperability of digital dental images, using the DICOM Standard as the backbone of the effort. ADA Working Group 12.1 was tasked with development of specifications and also with educating the dental profession concerning digital image interoperability. DICOM-related interoperability demonstrations are now a part of the ADA Annual Congress, in the form of seminar and as a noncommercial exhibit. PMID- 15551106 TI - [Shone's anomaly Fallbericht und Hintergrund]. AB - Shone's anomaly was first described in 1963 as a developmental complex of four potentially obstructive cardiac lesions including a supravalvular fibrous mitral ring, deformity of the mitral and/or subvalvular apparatus, subvalvular aortic stenosis and coarctation of the aorta. While paediatric patients with Shone's anomaly have been reported in the literature, only a few adult patients presenting with this anomaly have been described in the perioperative period. However, patients with an undiagnosed, incomplete form of Shone's anomaly might occasionally present for non-cardiac surgery as adults. In this case report we describe the anaesthetic management of an adult patient scheduled for a non cardiac operation, who suffered from Shone's anomaly that was unrecognised prior to the operation. PMID- 15551107 TI - Matrix metalloproteinases and their regulators are cardiovascular therapeutic targets. PMID- 15551108 TI - Mastrus ridibundus parasitoids eavesdrop on cocoon-spinning codling moth, Cydia pomonella, larvae. AB - Cocoon-spinning larvae of the codling moth, Cydia pomonella L. (Lepidoptera: Olethreutidae) employ a pheromone that attracts or arrests conspecifics seeking pupation sites. Such intraspecific communication signals are important cues for illicit receivers such as parasitoids to exploit. We tested the hypothesis that the prepupal C. pomonella parasitoid Mastrus ridibundus Gravenhorst (Hymenoptera: Ichneumonidae) exploits the larval aggregation pheromone to locate host prepupae. In laboratory olfactometer experiments, female M. ridibundus were attracted to 3 day-old cocoons containing C. pomonella larvae or prepupae. Older cocoons containing C. pomonella pupae, or larvae and prepupae excised from cocoons, were not attractive. In gas chromatographic-electroantennographic detection (GC-EAD) analyses of bioactive Porapak Q extract of cocoon-derived airborne semiochemicals, ten compounds elicited responses from female M. ridibundus antennae. Comparative GC-mass spectrometry of authentic standards and cocoon volatiles determined that these compounds were 3-carene, myrcene, heptanal, octanal, nonanal, decanal, (E)-2-octenal, (E)-2-nonenal, sulcatone, and geranylacetone. A synthetic 11-component blend consisting of these ten EAD-active compounds plus EAD-inactive (+)-limonene (the most abundant cocoon-derived volatile) was as effective as Porapak Q cocoon extract in attracting both female M. ridibundus and C. pomonella larvae seeking pupation sites. Only three components could be deleted from the 11-component blend without diminishing its attractiveness to M. ridibundus, which underlines the complexity of information received and processed during foraging for hosts. Mastrus ridibundus obviously "eavesdrop" on the pheromonal communication signals of C. pomonella larvae that reliably indicate host presence. PMID- 15551109 TI - [Suicide mortality among Turks in Germany]. AB - BACKGROUND: Suicide mortality is usually higher among men than among women. In socially disadvantaged groups, this relationship can be reversed. We investigated whether this is the case among Turkish migrants in Germany. MATERIAL AND METHODS: We calculated age- and sex-specific suicide rates for Turkish nationals residing in (West) Germany (1186 suicides) and for Germans aged below 65 years based on death registration data covering the period 1980-1997. RESULTS: Age-adjusted suicide rates among Turks were lower than among Germans (relative risk 0.3). Among Turkish girls and young women under 18 years, however, the relative risk compared to Germans was 1.8 (95% confidence interval 1.4-2.3). The male-to-female ratio in this age group was 3.2 among Germans and 0.6 among Turks. DISCUSSION: Overall, Turks living in Germany have a lower suicide mortality than Germans. Possible explanations include a high level of social coherence in the Turkish community and religious prohibitions. The high suicide rate among Turkish girls and young women could indicate the presence of social or cultural conflict situations. PMID- 15551110 TI - [Alzheimer's disease with secondary Parkinson's syndrome. Case report of a patient with dementia and Parkinson's syndrome after long-term occupational exposure to insecticides, herbicides, and pesticides]. AB - This case report describes long-term occupational exposure to agricultural insecticides, herbicides, and pesticides as possible environmental risk factors of Alzheimer's disease (AD) and Parkinson's syndrome in a 59-year-old man. Initially the patient complained about disturbances in concentration, mnestic deficits, and problems finding words. In the further course of the disease, he developed Parkinson's syndrome with predominant hypokinesia and rigor in addition to mild-to-moderate dementia. Low levels of beta-amyloid 1-42 were found in the CSF. Electroencephalography showed left frontotemporal theta waves. Cranial MRI revealed general brain atrophy with a maximum biparietally. In cerebral positron emission tomography, general hypometabolism was found with maxima biparietally and left frontally. The possible differential diagnosis of AD and Parkinson's syndrome is discussed. PMID- 15551111 TI - [Mental health care in developing countries: the example of Nigeria]. AB - The availability of mental health resources in most developing countries is poor due to scarcity of resources, competing health problems, and the low priority given to mental health issues. In Nigeria, modern psychiatric care is developing, but as yet most people have no access to it because they cannot afford the treatment. Moreover, treatment facilities are mainly concentrated in large urban centers. The cooperation between the authors of this article is aimed at improving the provision of psychiatric care for people living in Ibadan, Nigeria and rural areas close to the city. We report about the foundation of an association to provide help for mentally ill patients in Africa. PMID- 15551117 TI - National Osteoporosis Society 10th Scientific Conference. Harrogate, United Kingdom, 28 November - 1 December 2004. Abstracts. PMID- 15551112 TI - High-resolution mapping of the Rym4/Rym5 locus conferring resistance to the barley yellow mosaic virus complex (BaMMV, BaYMV, BaYMV-2) in barley (Hordeum vulgare ssp. vulgare L.). AB - Soil-borne barley yellow mosaic virus disease--caused by a complex of at least three viruses, i.e. Barley mild mosaic virus (BaMMV), Barley yellow mosaic virus (BaYMV) and BaYMV-2--is one of the most important diseases of winter barley in Europe. The two genes rym4, effective against BaMMV and BaYMV, and rym5, additionally effective against BaYMV-2, comprise a complex locus on chromosome 3HL, which is of special importance to European barley breeding. To provide the genetic basis for positional cloning of the Rym4/Rym5 locus, two high-resolution maps were constructed based on co-dominant flanking markers (MWG838/Y57c10 - MWG010/Bmac29). Mapping at a resolution of about 0.05% rec., rym4 has been located 1.07% recombination distal of marker MWG838 and 1.21% recombination proximal to marker MWG010. Based on a population size of 3,884 F2 plants (0.013% recombination) the interval harbouring rym5 was delimited to 1.49+/-0.14% recombination. By testing segmental recombinant inbred lines (RILs) for reaction to the different viruses at a resolution of 0.05% rec. (rym4) and 0.019% rec. (rym5), no segregation concerning the reaction to the different viruses could be observed. AFLP-based marker saturation for rym4, using 932 PstI+2/MseI+3 primer combinations only resulted in three markers with the closest one linked at 0.9% recombination to the gene. Two of these markers detected epialleles arising from the differential cytosine methylation of PstI sites. Regarding rym5, profiling of 1,200 RAPD primers (about 18,000 loci) and 2,048 EcoRI+3/MseI+3 AFLP primer combinations (about 205,000 loci) resulted in one RAPD marker and seven AFLP markers tightly linked to the resistance gene. Flanking markers with the closest linkage to rym5 (0.05% and 0.88% recombination) were converted into STS markers. These markers provide a starting point for chromosomal walking and may be exploited in marker-assisted selection for virus resistance based on rym5. PMID- 15551118 TI - Gene cloning, expression and functional characterization of a phosphopantetheinyl transferase from Vibrio anguillarum serotype O1. AB - Phosphopantetheinyl transferases (PPTases) catalyze the essential post translational activation of carrier proteins from fatty acid synthetases (FASs) in primary metabolism and polyketide synthetases (PKSs) and non-ribosomal polypeptide synthetases (NRPSs) in secondary metabolism. Bacteria typically harbor one PPTase specific for carrier proteins of primary metabolism (ACPS-type PPTases) and at least one capable of modifying carrier proteins involved in secondary metabolism (Sfp-type PPTases). Anguibactin, an important virulent factor in Vibrio anguillarum serotype O1, has been reported to be synthesized by a nonribosomal peptide synthetases (NRPS) system encoded on a 65-kb virulent plasmid pJM1 from strain 775 of V. anguillarum serotype O1, and the PPTase, necessary for the activation of the anguibactin-NRPS, is therefore expected to lie on the pJM1 plasmid. In this work, a putative PPTase gene, angD, was first identified on pEIB1 plasmid (a pJM1-like plasmid) from a virulent strain MVM425 of V. anguillarum serotype O1. A recombinant clone carrying complete angD was able to complement an Escherichia coli entD mutant deficient in Sfp-type PPTase. angD was overexpressed in E. coli and the resultant protein, AngD, was purified. Simultaneously, two carrier proteins involved in anguibactin-NRPS, ArCP and PCP, were overproduced in E. coli and purified. The purified AngD, PCP and ArCP were used to establish an in vitro enzyme reaction, and the PPTase activity of AngD was proved through HPLC analysis to detect the conversion of inactive carrier proteins to active carrier proteins in the reaction mixture. Co-expression of AngD with PCP or ArCP showed that AngD functioned well as a PPTase in vivo in E. coli, modifying PCP and ArCP completely. PMID- 15551121 TI - Loss of serotonin 5-HT2A receptors in the postmortem temporal cortex correlates with rate of cognitive decline in Alzheimer's disease. AB - RATIONALE: Previous studies have demonstrated reductions of serotonin 5-HT 2A receptors in the neocortex of Alzheimer's disease (AD) patients. However, it is unclear whether such losses play a role in the cognitive decline of AD. OBJECTIVES: To correlate neocortical 5-HT 2A receptor alterations with cognitive decline in AD. METHODS: Postmortem frontal and temporal cortical 5-HT 2A receptors were measured by [3H]ketanserin binding in aged controls as well as in a cohort of AD patients who had been longitudinally assessed for cognitive decline and behavioral symptoms. RESULTS: 5-HT 2A receptor densities in both regions were reduced in severely demented AD patients compared to age-matched controls. In the temporal cortex, this reduction also correlated with the rate of decline of Mini-Mental State Examination (MMSE) scores. The association between 5 HT 2A receptor loss and cognitive decline was independent of the effects of choline acetyltransferase (ChAT) activity and presence of behavioral symptoms. CONCLUSIONS: Our data suggest that loss of neocortical 5-HT 2A receptors may predict for faster cognitive decline in AD, and point to serotomimetics as potentially useful adjuvants to cholinergic replacement therapies. PMID- 15551129 TI - Desmoplastic infantile ganglioglioma: MRI and histological findings case report. AB - Desmoplastic infantile gangliogliomas (DIG) are rare intracranial tumors occurring during the 1st year of life. They arise invariably in the supratentorial region and have a great size at presentation, commonly involving more than one lobe. They are composed of a solid peripheral component of variable size, which involves the superficial cerebral cortex and the leptomeninges, and a large cystic part. Despite the great size at presentation and occasional mitotic activity in the variable undifferentiated component, this entity constitutes a distinct clinicopathological entity with benign prognosis. We hereby present the MRI and histological findings of two cases of DIG in infants aged 9 and 10 months, respectively. PMID- 15551131 TI - 68Ga-DOTANOC: a first compound for PET imaging with high affinity for somatostatin receptor subtypes 2 and 5. PMID- 15551133 TI - Intraosseous lipoma: retrospective analysis of 29 patients. AB - We reviewed 29 patients with intraosseous lipoma treated between 1985 and 2002. Mean patient age was 48 (20-75) years. According to Milgram's classification, ten cases were classified as stage I, 14 as stage II, and three as stage III. All patients were initially treated by curettage. In 11 cases an additional phenolization was performed. The average follow-up was 32 (6-208) months. At the last follow-up, none had any clinical or radiological signs of recurrence. The adequate treatment of a symptomatic intraosseous lipoma is curettage and bone grafting. In the current study, phenolization showed no added benefit. An asymptomatic intraosseous lipoma without impending pathological fracture can be treated conservatively. PMID- 15551134 TI - Fungal degradation of wood: initial proteomic analysis of extracellular proteins of Phanerochaete chrysosporium grown on oak substrate. AB - Two-dimensional (2-D) gel electrophoresis was used to separate the extracellular proteins produced by the white-rot fungus Phanerochaete chrysosporium. Solid substrate cultures grown on red oak wood chips yielded extracellular protein preparations which were not suitable for 2-D gel analysis. However, pre-washing the wood chips with water helped decrease the amount of brown material which caused smearing on the acidic side of the isoelectric focusing gel. The 2-D gels from these wood-grown cultures revealed more than 45 protein spots. These spots were subjected to in-gel digestion with trypsin followed by either peptide fingerprint analysis by matrix assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOF/MS) or by liquid chromatography (LC)/MS/MS sequencing. Data from both methods were analyzed by Protein Prospector and the local P. chrysosporium annotated database. MALDI-TOF/MS only identified two proteins out of 25 analyzed. This was most likely due to problems associated with glycosylation. Protein sequencing by LC/MS/MS of the same 25 proteins resulted in identification of 16 proteins. Most of the proteins identified act on either cellulose or hemicellulose or their hydrolysis products. Thus far no lignin peroxidase, Mn peroxidase or laccases have been detected. PMID- 15551135 TI - The yeast VPS genes affect telomere length regulation. AB - Eukaryotic cells invest a large proportion of their genome in maintaining telomere length homeostasis. Among the 173 non-essential yeast genes found to affect telomere length, a large proportion is involved in vacuolar traffic. When mutated, these vacuolar protein-sorting (VPS) genes lead to telomeres shorter than those observed in the wild type. Using genetic analysis, we characterized the pathway by which VPS15, VPS34, VPS22, VPS23 and VPS28 affect the telomeres. Our results indicate that these VPS genes affect telomere length through a single pathway and that this effect requires the activity of telomerase and the Ku heterodimer, but not the activity of Tel1p or Rif2p. We present models to explain the link between vacuolar traffic and telomere length homeostasis. PMID- 15551136 TI - Efficient plant regeneration from suspension cells of Allium cepa L. AB - Plant regeneration from calli of three cultivars of Allium cepa (Senshuki, O.Pki and Shojovaka) was investigated. Callus was induced on four variations of BDS medium containing different concentrations of 2,4-dichlorophenoxyacetic acid (2,4 D) and 6-benzyladenine (BAP). The regeneration frequency of calli of cvs. Senshuki and O.Pki subcultured on solid MS medium supplemented with BAP ranged from 50% to 80%; this frequency decreased to less than 30% after subculture in the dark in liquid BDS medium. By repeating the dark/light transitions of the culture protocol and by selecting for green cell clusters, we were able to increase the regeneration frequency to more than 80% in all three cultivars. These cell clusters maintained a high regeneration capacity in subsequent subcultures in the absence of light for 2 months. Most (97%) of the regenerated plantlets had a normal diploid karyotype (2 n=16) that was identical to that of the mother plants, although 3% of the regenerated plants of cv. Shojovaka had a tetraploid karyotype. PMID- 15551137 TI - Alteration of biomass and artemisinin production in Artemisia annua hairy roots by media sterilization method and sugars. AB - Transformed root cultures of Artemisia annua grown in autoclaved medium show large variations in biomass and artemisinin production regardless of the culture conditions or clonal type. However, using filter-sterilized sugars singly or in combination while holding the carbon level in the medium constant resulted in an unexpected variability in biomass production and artemisinin yield. Autoclaving results in variable hydrolysis of sucrose in the culture medium. Subsequent experiments using combinations of filter-sterilized sugars at a constant total carbon level in the medium showed a stimulation of artemisinin production by glucose. Growth in sucrose was equivalent to growth in fructose and significantly better than in glucose. These results suggest that sugars may be affecting terpenoid metabolism not only as carbon sources, but also as signal molecules. PMID- 15551138 TI - Down-regulation of Ku 70 and Ku 80 mRNA expression in transitional cell carcinomas of the urinary bladder related to tumor progression. AB - DNA-dependent protein kinase (DNA-PK) containing the regulatory subunits Ku 70 and Ku 80 plays a prominent role in the repair of double-stranded DNA breaks by a nonhomologous end-joining pathway maintaining genomic stability. In an attempt to elucidate the significance of the DNA-PK complex for human urothelial carcinogenesis, the expression of Ku 70 and Ku 80 was studied in 71 transitional cell carcinomas (TCC) of the urinary bladder of various grades and stages, and in relation to lifestyle and occupational bladder cancer risk factors. To analyse the mRNA expression of Ku 70 and Ku 80, real-time quantitative reverse transcription-polymerase chain reaction was used and the protein expression assessed by immunohistochemistry. Advanced high-grade, high-stage TCC expressed the mRNA of Ku 70 and Ku 80 at a lower level than superficial low-grade, low stage carcinomas, suggesting down-regulation of the Ku system to be associated with progression of bladder cancer from a low to a high malignant potential. The protein expression of Ku 70 and Ku 80 was closely related and decreased consistently with increasing grades and stages, paralleling the expression of the mRNA. Among hazardous environmental bladder cancer risk factors, heavy consumption of coffee was associated with a twofold decreased Ku 70 and Ku 80 mRNA expression, whereas tobacco smoke did not substantially affect the activity of the Ku system, except for a trend towards a dose-response relationship in the expression of Ku 70 mRNA. There is some evidence that exposure to polycyclic hydrocarbons, paints and lacquer, and stone dust may modify the expression of Ku 70 mRNA. Although the underlying molecular genetic pathways are not yet clearly understood, our data indicate that down-regulation of the Ku system promotes progression of urothelial carcinogenesis to a more malignant and aggressive clinical behavior, presumably as a result of an impaired capacity for DNA repair. PMID- 15551139 TI - Inner calvarial erosion from traumatic subdural haematoma in infancy. AB - CASE REPORT: This is a report of a case of a subdural haematoma in infancy of possible non-accidental aetiology with raised pericerebral pressure, which we postulate has eroded the inner table of the cranial bones and resulted in leakage of marrow precursor cells into the extradural space. RESULT: Subdural tapping via the fontanelle has created a channel allowing subsequent ingress of nucleated red cell precursors into the subdural space. This addition to the subdural collection has prolonged its course necessitating subduro-peritoneal shunting. PMID- 15551142 TI - Neutrophils infiltrating ultraviolet B-irradiated normal human skin display high IL-10 expression. AB - Exposure to an erythemal dose of ultraviolet B (UVB) is known to induce interleukin (IL-10) expression in human skin. It is generally believed that this IL-10 is predominantly expressed by CD11b+ HLA-DR+ macrophages that infiltrate the UVB-exposed skin. This cytokine is presumed to contribute to the immunosuppressive effects of UVB by inhibiting cell-mediated immune responses. We recently demonstrated that neutrophils, which also invade UVB-irradiated skin, express CD11b and HLA-DR as well. In addition, we showed that the presence of these neutrophils affects T-cell responses in primary T-cell cultures derived from UVB-exposed skin. Since neutrophils invade UVB-exposed skin and, like macrophages, express CD11b and HLA-DR, we sought to determine whether neutrophils represent another source of IL-10. Skin biopsies were obtained from four healthy volunteers before and 2 days after exposure to four minimal erythema doses of UVB. A series of immunohistochemical double-staining procedures using the following markers was performed: IL-10, CD11b, HLA-DR, CD36, neutrophil elastase, and CD66b. As expected IL-10 could be detected in CD11b+ HLA-DR+ CD36+ macrophages in the epidermis and dermis of UVB-exposed skin. Surprisingly, the majority of the abundant IL-10 expression was found in CD11b+ HLA-DR+ elastase+ CD66b+ neutrophils. Cytospin preparations from dermal cell suspensions confirmed the IL-10 expression by neutrophils displaying characteristic multilobular nuclei. Thus, neutrophils in UVB-exposed skin express IL-10 and should be recognized as active coplayers in the creation of the UVB-induced immunosuppressive microenvironment. PMID- 15551143 TI - Incarceration of the retroverted gravid uterus: the key to successful treatment. AB - INTRODUCTION: Incarceration of the retroverted gravid uterus is rare, and it is difficult to make a diagnosis, and, therefore, difficult to treat. We present two cases of incarceration of the retroverted gravid uterus. CASE REPORT: Two pregnant women presented with urinary retention at 14 weeks' gestation. We tried to push the gravid uterus upward, but in the end the two women underwent cesarean section. In one of the women it was possible to relieve incarceration of the retroverted gravid uterus. CONCLUSION: There was no sign of endometriosis in these pregnancies, which is important to evaluate when pregnant women have a complication. PMID- 15551144 TI - Malignant transformation of an ovarian mature cystic teratoma: computed tomography findings. AB - INTRODUCTION: The malignant transformation of an ovarian mature cystic teratoma is rare. We presented such a case of a 46-year-old woman with characteristic computed tomography (CT) findings. CASE REPORT: The patient had suffered from fullness and tenderness of the lower abdomen for one year. A high serum CA-125 level was noted. Computed tomography images showed a left adnexal mass with fat, calcification and soft tissue components, and areas of invasion through the teratoma wall. OUTCOME: The histologic diagnosis was compatible with a squamous cell carcinoma arising from a mature cystic teratoma of the ovary. PMID- 15551152 TI - Ultrastructural study of echinocytes induced by poly (ethylene glycol) cholesterol. AB - Poly (ethylene glycol)-cholesterol (PEG-Chol) consists of a hydrophilic PEG and hydrophobic cholesterol moiety. When PEG-Chol was applied to erythrocytes, the reagent quantitatively induced protrusions by exclusively distributing in the outer monolayer of the membrane. This kind of response has been regarded as a general response that reduces the stress of expansion of the outer monolayer. However, the relationship between the membrane architecture and the distribution of such molecules is unknown. In this study, we examined the distribution of tagged PEG-Chol along the shape change pathway. The echinocytic shape was initiated by the initial formation of bumps on the rim of the discoid, which subsequently elongated as protrusions. These protrusions contained aggregates of granular structures, which appeared to accommodate the increase in the outer monolayer area. At higher concentrations, PEG-Chol further induced sphero echinocytosis that resulted in numerous branched protrusion processes. We found that PEG-Chol was exclusively distributed in these protrusions and, in particular, accumulated at the tips. These results suggested that externally intercalated PEG-Chol was sequestrated from erythrocytes as membrane protrusions through an as-yet-unknown mechanism. PMID- 15551153 TI - Production and characterisation of cell- and tissue-specific monoclonal antibodies for the flatworm Macrostomum sp. AB - Monoclonal antibodies (mABs) against various cell types of the basal free-living flatworm Macrostomum sp. were produced by immunising Balb/c mice with cell suspensions of disintegrated animals. We identified 360 positive supernatants with specific staining of various tissues, cell types, patterns or structures. Here we report immunocytochemical characterisation, histological stainings and isotyping of 11 mABs specific for muscle cells (MMu-1, MMu-2, MMu-3, MMu-4), digestive and prostate glands (MDr-1 and MDr-2, MPr-1), epidermal cells (MEp-1), the ventral nerve cord including neuron clusters (MNv-1), gastrodermal cells (MDa 1) and spermatids (MSp-1). Confocal microscopy, histological techniques, electron microscopy and immunoblotting were applied to demonstrate stainings in juveniles, adults, starved or well-fed animals. Considering the current lack of specific markers the obtained mABs will be particularly helpful studying embryonic and postembryonic development, pattern formation, cell differentiation, regeneration and reproductive allocation in Macrostomum sp., and possibly other basal flatworms. The small size, ease of culturing, short generation time, transparency and the basal phylogenetic position specify Macrostomum sp. as a suitable model organism for comparative analyses within Platyhelminthes and to Drosophila and C. elegans. PMID- 15551155 TI - The distribution of P2X5 purinergic receptors in the enteric nervous system of mouse. AB - The distribution of the P2X(5) purinoceptor in the enteric nervous system of the mouse was studied by immunohistochemistry. P2X(5) receptor immunoreactivity was widely distributed in myenteric and submucosal plexuses throughout the gastrointestinal tract. In myenteric plexuses, immunoreactivity for the P2X(5) receptor was observed in nerve fibres that enveloped ganglion cell bodies, and possibly on glial cell processes. P2X(5) receptor immunoreactivity was colocalised with vasoactive intestinal peptide and surrounded ganglion cells that contained calretinin, calbindin or nitric oxide synthase. In the submucous plexus, P2X(5) receptor immunoreactivity occurred throughout the cytoplasm and on the surface membranes of the nerve cells. Double-labelling studies showed that 22%, 9%, 6% and 68% of P2X(5) receptor-immunoreactive neurones were also immunoreactive for calretinin, calbindin, nitric oxide synthase and vasoactive intestinal peptide, respectively. Thus, the P2X(5) receptor subunit is expressed in specific functional groups of neurones. P2X(2) and P2X(3) receptors were also present in the mouse enteric plexuses but no immunoreactivity for P2X(1), P2X(4) or P2X(6) receptors was found. PMID- 15551157 TI - The role of body position and gravity in the symptoms and treatment of various medical diseases. AB - Postural medicine studies the effects of gravity on human body functions and the ability to influence various diseases by changing the body's position. Orthostasis requires numerous cardiovascular and neurohumoral adaptations to prevent hypotension and a resulting decrease in cerebral perfusion. Sitting upright or in a semi-sitting position reduces venous return in patients with heart failure, intracranial pressure in patients with intracranial hypertension, intraocular pressure in glaucoma patients and may decrease gastro-oesophageal reflux. A left recumbent posture also decreases reflux. A right lateral position results in a lower sympathetic tone than lying on the left side and is beneficial in patients with heart failure or after an infarction without bradycardia. A 40 to 70% decreased prevalence of the sudden infant death syndrome has been observed since the recommendation to avoid laying infants to sleep in a prone position. Sleeping in a supine posture increases the severity of sleep apnoea compared to a lateral position. In patients with acute respiratory distress syndrome, a prone position can rapidly improve blood oxygenation. Idiopathic oedema, orthostatic proteinuria, intradiscal pressure and venous circulation in legs are improved in the decubitus position, whereas arterial flow is reduced. Health risks due to microgravity and prolonged bed rest, such as osteoporosis, venous thrombosis or pressure sores, are discussed. PMID- 15551156 TI - Strong linkage disequilibrium of a HbE variant with the (AT)9(T)5 repeat in the BP1 binding site upstream of the beta-globin gene in the Thai population. AB - A binding site for the repressor protein BP1, which contains a tandem (AT)x(T)y repeat, is located approximately 530 bp 5' to the human beta-globin gene (HBB). There is accumulating evidence that BP1 binds to the (AT)9(T)5 allele more strongly than to other alleles, thereby reducing the expression of HBB. In this study, we investigated polymorphisms in the (AT)x(T)y repeat in 57 individuals living in Thailand, including three homozygotes for the hemoglobin E variant (HbE; beta26Glu-->Lys), 22 heterozygotes, and 32 normal homozygotes. We found that (AT)9(T)5 and (AT)7(T)7 alleles were predominant in the studied population and that the HbE variant is in strong linkage disequilibrium with the (AT)9(T)5 allele, which can explain why the betaE chain is inefficiently synthesized compared to the normal betaA chain. Moreover, the mildness of the HbE disease compared to other hemoglobinopathies in Thai may be due, in part, to the presence of the (AT)9(T)5 repeat on the HbE chromosome. In addition, a novel (AC)n polymorphism adjacent to the (AT)x(T)y repeat (i.e., (AC)3(AT)7(T)5) was found through the variation screening in this study. PMID- 15551158 TI - Directly observed therapy for tuberculosis in a low prevalence region: first experience at the Tuberculosis Dispensary in Lausanne. AB - AIM: Evaluation of first experience of the directly observed therapy (DOT) programme for tuberculosis introduced in the Canton of Vaud in 1997. METHOD: Retrospective analysis of tuberculosis patients included in a DOT programme from October 1997 to March 2000 under the supervision of the TB Dispensary in Lausanne. RESULTS: 54 patients were included, 49 of whom were new cases and 5 relapses. 70% were asylum seekers and illegal immigrants. The indications for DOT were immigrant status, social problems, and physical or psychiatric comorbidities. Treatment was fully supervised in 67% and partially in 33%. The outcome was favourable (cure or treatment completion) in 88.9% and unfavourable in 11.1%. A similar success rate was observed in full and partial DOT and there was no difference in success rates between the various structures where DOT was administered. By comparison, the success rate in a historical group from the same institution was 70% in 1990. The biggest problem was communication with the patients and within the team. CONCLUSION: Treatment of tuberculosis under DOT in the Canton of Vaud resulted in improvement of the treatment success rate. PMID- 15551159 TI - Islets secretory capacity determines glucose homoeostasis in the face of insulin resistance. AB - BACKGROUND: Debate surrounds the relative importance of impaired beta-cell secretory function versus insulin resistance in type 2 diabetes. We therefore defined insulin secretion and sensitivity in patients with impaired glucose homoeostasis of varying aetiology and duration. METHODS: 126 consecutive patients undergoing an oral glucose tolerance test (OGTT) between 1999 and 2003 were included. Whole-body insulin sensitivity index (ISI) and insulinogenic index derived from the OGTT were determined in 32 healthy controls, 65 type 2 diabetic patients, 15 patients with acromegaly, 10 patients with insulinoma, and 4 patients with HAIRAN syndrome. RESULTS: Median ISI (quartiles Q25-Q75) of healthy controls and of patients with insulinoma were similar (3.5 [2.8-5.6] and 3.2 [1.7 4.2] respectively) but significantly decreased in patients with acromegaly, type 2 diabetes, and HAIRAN syndrome (2.8 [1.8-3.3], 1.9 [1.4-3], and 0.8 [0.6-1.3] respectively). Despite the decrease in ISI, patients with HAIRAN syndrome and acromegaly maintained normal glucose tolerance by adapting insulin secretion as reflected in the insulinogenic index (106.5 [90.4-127.5] and 49 [24.4-89] in HAIRAN and acromegaly respectively, versus 46.9 [27.3-66.7] in controls). In contrast, type 2 diabetic patients failed to adapt and displayed severely hampered insulin secretion (insulinogenic index of 7.6 [3.8-14.7]). Furthermore, the level of the insulinogenic index correlated significantly with duration of diabetes and HbA1c, which was not the case for the ISI. Insulinoma patients had a decreased insulinogenic index (38.7 [32-83.8]), leading to impaired glucose tolerance despite normal ISI. CONCLUSIONS: The data are compatible with the notion that beta-cell function rather than insulin sensitivity determines the evolution of hyperglycaemia. PMID- 15551160 TI - Comparison of the efficacy of two months of treatment with co-trimoxazole plus doxycycline vs. co-trimoxazole plus rifampin in brucellosis. AB - BACKGROUND: To compare the efficacy of two different regimens in treatment of brucellosis. METHODS: This randomised clinical trial study was conducted on 280 patients with brucellosis in Babol, Iran, from April 1999 to January 2002. One of the following two regimens was randomly prescribed for two months: co-trimoxazole plus doxycycline (CD group) and co-trimoxazole plus rifampin (CR group). RESULTS: 140 patients with the mean age of 35.56 (16.2) years, and 140 patients with the mean age of 31.39 (18) years, were treated with co-trimoxazole plus doxycycline and co-trimoxazole plus rifampin, respectively. Clinical manifestations and laboratory test results were similar in both groups (p >0.05), except in sex and clinical types (p <0.05). Failure of treatment was seen in 10 (7.1%) and 23 (16.4%) cases treated in the CD group and CR group, respectively (95% CI, 0.174 to 0.862; OR = 0.387; p = 0.020). Relapse was seen in 12 cases (8.6%) treated in the CD group and in 14 cases (10%) treated in the CR group (95% CI, 0.365 to 1.87; OR = 0.826; p = 0.646). Failure of treatment plus relapse was seen in 22 (15.7%) and 37 (26.4%) cases treated in the CD group and CR group, respectively (95%CI, 0.278 to 0.929, OR = 0.508; p = 0.028). Risk for developing of failure of treatment and relapse in patients treated with co-trimoxazole plus rifampin was 1.96 times higher than those treated with co-trimoxazole plus doxycycline. Among the relapsed patients, 18 (69.2%) cases occurred within 6 months after completion of therapy, and most of them in uncomplicated patients. CONCLUSION: Using two months of treatment, the efficacy of co-trimoxazole plus doxycycline is better than co-trimoxazole plus rifampin. PMID- 15551161 TI - EORTC-NCI-AACR--16th Symposium. Molecular targets and cancer therapeutics. 28 September-1 October 2004, Geneva, Switzerland. PMID- 15551162 TI - EORTC-NCI-AACR--16th Symposium. Kinase inhibitors. 28 September-1 October 2004, Geneva, Switzerland. PMID- 15551163 TI - EORTC-NCI-AACR--16th Symposium. Highlights. 28 September-1 October 2004, Geneva, Switzerland. PMID- 15551164 TI - Monoclonal Antibodies in Cancer--Fourth International Congress. 3-6 September 2004, Colorado Springs, CO, USA. PMID- 15551165 TI - Monoclonal Antibodies in Cancer--Fourth International Congress. EGFR/HER1 antibodies. 3-6 September 2004, Colorado Springs, CO, USA. PMID- 15551166 TI - Monoclonal Antibodies in Cancer--Fourth International Congress. HER2/neu and other antibodies. 3-6 September 2004, Colorado Springs, CO, USA. PMID- 15551167 TI - European Association for the Study of Diabetes--40th Annual Meeting. 5-9 September 2004, Munich, Germany. PMID- 15551168 TI - Transplantation Society--XXth International Congress. 5-10 September 2004, Vienna, Austria. PMID- 15551169 TI - Pain Management 2004--Analyzing regulatory, commercial and pharmaceutical practices. 8-9 September 2004, London, UK. PMID- 15551170 TI - European Peptide Society--28th Symposium and Third International Peptide Symposium. Peptides--bridges between disciplines. 5-10 September 2004, Prague, Czech Republic. PMID- 15551171 TI - European Society for Dermatological Research--34th Annual Meeting. 9-11 September 2004, Vienna, Austria. PMID- 15551172 TI - Pharmaceutical Technology Conference--12th International Meeting. Problems and problem solving strategies in pharmaceutical formulation and drug delivery. 12-15 September 2004, Istanbul, Turkey. PMID- 15551173 TI - Pharmacology--Fourth Croatian Congress. 15-18 September 2004, Split, Croatia. PMID- 15551174 TI - Drug Discovery Technology Asia-Pacific 2004--IBC's Second Annual Conference and Exhibition. Drug discovery to manufacturing: global partnering and new science. 18-20 October 2004, Mumbai, India. PMID- 15551175 TI - Drug Discovery Technology Asia-Pacific 2004--IBC's Second Annual Conference and Exhibition. Drug discovery and development track highlights. 18-20 October 2004, Mumbai, India. PMID- 15551176 TI - Conus toxins: targets and properties. AB - The venoms from Conus snails are rich in peptides with potent specificity for mammalian receptor sites. Each venom typically contains up to 100 conopeptides, and with approximately 500 species of Conus snail, the number of active peptides is considerable. The receptor sites targeted appear to be mostly linked to ion channels, with voltage-gated, ligand-gated and G-protein linked sites identified. Both the central and peripheral nervous system present possible physiological targets for therapeutic products derived from the venoms, although the molecules in the most advanced development target the central nervous system. In turn, this presents problems of bioavailability, while the potency is a potential source of toxicity. PMID- 15551177 TI - Pharmacognosy and reverse pharmacognosy: a new concept for accelerating natural drug discovery. AB - Combinatorial chemistry and high-throughput screening (HTS) have led to the identification of numerous agents that are active and selective in vitro. Identifying drugs that are active in vivo, however, remains a challenge. Traditional medicinal cures based on natural materials have proven useful for many populations worldwide, representing huge and disperse tracts of knowledge that are sometimes neglected in Western research due to differences in the concepts of illness. In this review we introduce a new approach, termed 'reverse pharmacognosy' (from diverse molecules to plants), which can be coupled with pharmacognosy (from biodiverse plants to molecules). Reverse pharmacognosy utilizes new techniques, such as HTS, virtual screening and a knowledge database containing the traditional uses of plants. Integrating pharmacognosy and reverse pharmacognosy in the research process may provide an efficient and rapid tool for natural drug discovery. PMID- 15551178 TI - Idraparinux sodium. Sanofi-Aventis. AB - Idraparinux sodium, a long-acting anti-Factor Xa synthetic pentasaccharide, is under development by Sanofi-Aventis for the potential prevention and treatment of venous thromboembolic events in patients with deep vein thrombosis or pulmonary embolism. PMID- 15551179 TI - Surgical aspects and outcome of delayed tethered cord release. AB - Out of a total of 550 children followed up at our spina bifida center, we report on 81 patients who were reoperated upon for secondary tethered cord syndrome between 1993 and 2000. In four cases with preceding severe progressive scoliosis, untethering was followed by surgical correction and stabilization of curvatures. In 77 patients, the indication for surgery was based on late progressive neurological deterioration. The current clinical relevance of competing etiologic factors such as symptomatic Chiari malformation, hydromyelia, and shunt dysfunction, requiring different clinical management, had been previously carefully excluded. The children underwent magnetic resonance imaging (MRI) or myelo-computed tomography (m-CT) to identify the morphologic extent of tethering and any associated spinal malformations such as dermoid tumors (19 cases) or diastematomyelia (9 cases). Surgery became faster and safer through bilateral dural incision, undercutting arachnoid adhesions along the tethered area, although this procedure increased the need for dural grafting. Complete release of the conus medullaris and cauda equina was achieved in a total of 75 cases (93 %) including those who had undergone prophylactic surgery. A mean follow-up of 4.8 years in 77 patients operated upon for late progressive neurological deterioration confirmed stabilization of presenting symptoms in 65 cases (84 %) with 20 of them (26 %) even showing significant improvement. In 12 patients (16 %), including all cases of incomplete untethering (n = 4), there was further deterioration. PMID- 15551180 TI - The neurosurgeon on duty -- quality and quantity of acute care. AB - OBJECTIVE: An important part of the daily routine in neurosurgery is the treatment of emergency room admissions, acute cases from other departments or from outside hospitals. This acute care is not normally included in performance figures or budget management nor analysed scientifically with respect to quantity and quality of care provided by neurosurgeons. METHOD: Over a one-year period, all acute care cases managed by two neurosurgical on-call teams in Hannover (Northern Germany, 522 000 inhabitants) were recorded prospectively on a day-by day basis. A large database of 1 819 entries was created and analysed using descriptive statistics. RESULTS: The minimum incidence of neurosurgical acute care cases was estimated to be 75-115/100 000 inhabitants/year. This corresponds to a mean of approximately 6 per day. The majority of patients was admitted after 5 p. m. and on weekends. Only 30 % of cases came directly via the emergency room. The fate of 70 % of patients depended initially on the "neurosurgical qualification" of primary care doctors and here deficits existed. Over one year the additional workload from acute care amounted to 1 000 unplanned admissions, 900 acute imaging procedures and almost 600 emergency operations. CONCLUSION: The current policy in public health which includes cuts in resources, transport facilities and manpower is not compatible with the demonstrated extent of acute neurosurgical care. In addition to routine elective work, a high number of extra admissions, evening or night-time surgery, and imaging procedures has to be carried out. These conclusions hold a special importance if health authorities wish to not just maintain present standards but to improve existing deficits. PMID- 15551181 TI - Fusion of MRI and CT with subdural grid electrodes. AB - In patients with drug-resistant focal epilepsies subdural grid electrodes may be implanted to determine the seizure onset zone and eloquent cortex areas. Since the spatial relationship of the grid to the underlying brain is poorly visualized on MRI, we co-registered MRI before and CT after implantation of subdural grid electrodes. In this study we sought an appropriate algorithm to combine both imaging modalities. We compared six different co-registration algorithms including surface-oriented, mutual information-based and landmark-based methods. The resulting overlay matrices were analyzed by calculating rotational and translational shifts and by judging co-registered MRI and CT scans visually. A brain surface oriented method had the lowest rotational (axial 0.7 +/- 0.6 degrees; coronal 1.7 +/- 1.1 degrees; sagittal 1.9 +/- 1.8 degrees) and translational shifts (3.7 +/- 1.3 mm). It was judged visually to be the best, had a low intra- and inter-observer variability, and lasted approximately 15 minutes. This algorithm is recommended when co-registering MRI before and CT after implantation of subdural grid electrodes. Skin-, voxel-, and landmark-based algorithms are less accurate, which is most likely due to postsurgical deformation of extra- and intracranial soft tissue. PMID- 15551182 TI - Spontaneous, multiple meningiomas. AB - OBJECT: The aim of this study is to discuss the management strategy of patients with multiple meningiomas. METHODS: We describe 35 patients with multiple meningiomas. No patients carried stigmata of neurofibromatosis. All patients were studied using MR or CT after surgical treatment. RESULTS: All 35 patients received surgical treatment. A total of 40 operations was performed. The resection of 4 meningiomas was subtotal and all other tumors were radically resected. We had excellent results with reversible operative complications in only three patients. There were no recurrences. CONCLUSIONS: We suggest that surgery is suitable for the treatment of multiple meningiomas with the following characteristics: symptomatic meningioma, asymptomatic meningioma greater than 3 cm in size and surgically accessible, and asymptomatic expanding tumor. In patients with multiple tumors, each tumor must be treated as an individual entity. The mere presence of multiple tumors does not justify their removal. PMID- 15551183 TI - A less invasive approach technique for operative treatment of lumbar canal stenosis. Technique and preliminary results. AB - The aim of this prospective study is to evaluate the efficacy of a less invasive approach for the operative treatment of lumbar canal stenosis. Using transmuscular dilatation this technique minimises surgical trauma. Through a working channel with a 11 mm outer diameter and under operative microscope guidance, decompression of the neural elements can be achieved. MATERIAL AND METHODS: From November 1998 to December 2001 38 consecutive patients with a mean age of 73.2 years were operated upon using this technique. The study included 56 lumbar segments. The mean follow up was 32 (18-55) months. The Visual Analogue Scale (VAS) for back and leg pain together with the Oxford Claudication Score (OCS) were used to assess the functional results. RESULTS: The average operating time for one level decompression was 74 minutes and the average blood loss was 32 ml/patient. Two patients (5.2 %) required revision by open surgery. The OCS improved from 29.4 preoperatively to 16.9 postoperatively (p < 0.001). VAS for back pain and leg pain also showed a significant improvement (p < 0.001). CONCLUSIONS: This less invasive approach is effective in decompressing lumbar canal stenosis. It has the advantage of early mobilisation and a short hospital stay, especially in elderly patients (mean age here: 73.2 years). PMID- 15551184 TI - Primary peripheral T-cell lymphoma of the central nervous system. AB - A 47-year-old man was admitted to our hospital with a nine-week history of visual disturbance, headache, disorientation and facial nerve palsy. Serial cranial magnetic resonance imaging (MRI) revealed progressive bilateral occipital edema with hemorrhage and meningeal involvement. There were no hints of hereditary or acquired immunodeficiency. Laboratory examination for bacterial and viral causes was negative. Open brain biopsy revealed primary central nervous system lymphoma of the extraordinary rare so-called "peripheral" T-cell type. The further course was fatal; the patient died 10 weeks after the onset of symptoms from tumor progression before planned chemotherapy could be started. CONCLUSION: If primary central nervous system lymphoma (PCNSL) is suspected, brain biopsy -- either open biopsy or stereotactic biopsy -- should be performed straight away to enable a rapid start of chemotherapy and/or radiotherapy. Peripheral T-cell lymphoma was highly aggressive in this case leading to the patient's death within several weeks. PMID- 15551185 TI - The diaphragma sellae meningioma -- a rare differential diagnosis of non functioning pituitary adenoma. AB - It is important to differentiate the rare Diaphragma sellae meningioma from the quite common hormonally inactive pituitary adenoma, especially with regards to the surgical approach. The differentiation could, however, be problematic. We report the case of a 66-year-old woman with a Diaphragma sellae meningioma, which became apparent through a visual field and visual acuity disorder. Successful radical tumor extirpation was achieved via the transsphenoidal approach, since the space-occupying lesion was initially thought to be a hormonally inactive pituitary macroadenoma. However, histological investigation confirmed a meningioma. A careful retrospective analysis of the MRI revealed the possibility of a meningioma having been overlooked, since not all distinguishing features of this lesion were manifested. These criteria, as well as the principles of the surgical approach for treating sellar space-occupying lesions are discussed. PMID- 15551186 TI - Prognostic value of improved intraoperative motor evoked potentials. A case report. AB - This unusual case involves intraoperative registration of improved motor evoked potentials accompanied by postoperative improvement of the motor status. CASE REPORT: A 51-year-old patient undergoing surgery for a right postcentral glioma was first submitted to phase reversal of somatosensory potentials for intraoperative localization of the central sulcus. During subsequent monopolar electric stimulation of the precentral gyrus, motor evoked potentials (MEPs) could not be recorded initially but only following extirpation of the tumor fraction in the central sulcus. The amplitudes of the registered potentials increased during the further clinical course. The preoperative arm-preponderant hemiparesis already improved on the first postoperative day. CONCLUSION: This intraoperative electrophysiological observation correlated with the postoperative neurological status, thus documenting functional recovery of the motor system through the intervention. PMID- 15551187 TI - [Incentive allocation of resources within medicine--are the psychosocial disciplines the losers?]. PMID- 15551188 TI - [Patterns of neuronal activity related to emotional stimulation in alexithymia]. AB - In alexithymia a frontal dysfunction is supposed to be a neurobiological correlate. This study focuses on distorted patterns of neuronal activity evoked by emotional stimuli in alexithymics and controls. Out of hospitalised patients with psychosomatic diseases 8 patients with a high score (HA) and 8 with a low one (NA) on the TAS-20 were investigated with fMRI during emotional stimulation which included pictures evoking anxiety and disgust as well as neutral illustrations. As response to negative affect arousing visual stimulation HA in comparison to NA showed a lower activation in the right medial prefrontal cortex and in the right amygdala. The results were significant for the emotion disgust. The results support the existence of a complex central feedback circuit consisting of regions of the prefrontal cortex and limbic structures to process negative affects. Hypothetically a fundamental factor for the emergence of alexithymic traits is an inhibiting process between affect processing (e. g. medial prefrontal cortex, anterior cingulum) and affect generating structures (e. g. amygdala). Furthermore our findings confirm the hypothesis that alexithymia is a disorder of higher cerebral function. PMID- 15551189 TI - [Factors influencing psychotherapeutic treatment outcome of various syndromes]. AB - The authors investigated specific and unspecific factors influencing the psychotherapeutic treatment of various syndromes using a questionnaire which systematically replicated the Consumer Reports Study performed in the USA in 1994. The authors were particularly concerned with the degree to which certain psychotherapeutic methods - psychoanalysis, depth psychology-based psychotherapy and behavioral therapy - produced differing results following treatment of syndromes. Using cluster-analysis, two groups of syndromes could be distinguished: Patients with depressive symptoms, stress-related disorders and/or relationship problems (depression-group) and patients with anxiety disorders and/or eating-related disorders (anxiety-group). With the help of cart-analysis (Classification and Regression Trees) it was possible to identify factors influencing the improvement of symptoms. The method of treatment had not a specific effect on the improvement of symptoms. In both groups the most important predictor was the length of treatment. Furthermore in the depression group the sex of the patients and a possible restriction of the treatment by the health insurance companies influenced the treatment results and in the anxiety group the frequency of treatment and the age of the patients. PMID- 15551190 TI - [Music therapy in oncology--concepts and review]. AB - The following article is addresses the applications and functions of music therapy in the acute and rehabilitative phases of treatment of adult cancer patients. It is based on a literature review with a focus on the state of empirical research in the oncology sector of music therapy and consequences for the concepts of music therapy. First the basic aspects of music therapy treatment are explained to clarify the different research methods and to examine the special demands of oncology patients. Furthermore, a brief summary of the approaches of music therapy research in the most renowned educational institutions in Germany are outlined. The results of the studies and case histories are summarized and evaluated and provide the basis for the conclusions and recommendations for the music therapeutic practice in oncology. PMID- 15551192 TI - Psychotropic effects of COX-2 inhibitors--a possible new approach for the treatment of psychiatric disorders. AB - Cyclooxygenase-2 (COX-2)--constitutively expressed in the CNS--is suggested to have an important functional role in the CNS. COX-2 interacts with neurotransmitters such as acetylcholine, serotonin, and glutamate, but is also involved in the regulation of immune system and in inflammation in the central nervous system (CNS) via effects of prostaglandins, in particular prostaglandin E (2). Recently, a role for the new generation of selective COX-2 inhibitors in the treatment of psychiatric disorders is discussed. Until now, COX-2 inhibitors have failed to show therapeutic effects in Alzheimer's disease, but studies from basic research point to a possible effect on cognition. A clinical effect of the COX-2 inhibitor celecoxib on cognition was observed in schizophrenic patients. The therapeutic effect of celecoxib add-on treatment to the atypical antipsychotic risperidone, however, is not restricted to cognition. A general effect on symptoms of schizophrenia was observed, which supports the view that an immunological/inflammatory process is involved in the pathogenesis of schizophrenia. In depression, however, signs of inflammation have been described since many years. Although results of clinical studies with COX-2 inhibitors in depression are still lacking, clinical improvement of a depressive syndrome has been observed in patients, who have been treated with rofecoxib due to other indications. These preliminary clinical data are encouraging for therapeutic effects of the selective COX-2 inhibitors in psychiatric disorders, although these effects have to be confirmed in bigger clinical studies. PMID- 15551191 TI - The AGNP-TDM expert group consensus guidelines: therapeutic drug monitoring in psychiatry. AB - Therapeutic Drug Monitoring (TDM) is a valid tool to optimise pharmacotherapy. It enables the clinician to adjust the dosage of drugs according to the characteristics of the individual patient. In psychiatry, TDM is an established procedure for lithium, some antidepressants and antipsychotics. In spite of its obvious advantages, however, the use of TDM in everyday clinical practice is far from optimal. The interdisciplinary TDM group of the Arbeitsgemeinschaft fur Neuropsychopharmakologie und Pharmakopsychiatrie (AGNP) has therefore worked out consensus guidelines to assist psychiatrists and laboratories involved in psychotropic drug analysis to optimise the use of TDM of psychotropic drugs. Five research-based levels of recommendation were defined with regard to routine monitoring of plasma concentrations for dose titration of 65 psychoactive drugs: (1) strongly recommended, (2) recommended, (3) useful, (4) probably useful and (5) not recommended. A second approach defined indications to use TDM, e. g. control of compliance, lack of clinical response or adverse effects at recommended doses, drug interactions, pharmacovigilance programs, presence of a genetic particularity concerning the drug metabolism, children, adolescents and elderly patients. Indications for TDM are relevant for all drugs either with or without validated therapeutic ranges. When studies on therapeutic ranges are lacking, target ranges should be plasma concentrations that are normally observed at therapeutic doses of the drug. Therapeutic ranges of plasma concentrations that are considered to be optimal for treatment are proposed for those drugs, for which the evaluation of the literature demonstrated strong evidence. Moreover, situations are defined when pharmacogenetic (phenotyping or genotyping) tests are informative in addition to TDM. Finally, practical instructions are given how to use TDM. They consider preparation of TDM, analytical procedures, reporting and interpretation of results and the use of information for patient treatment. Using the consensus guideline will help to ensure optimal clinical benefit of TDM in psychiatry. PMID- 15551193 TI - Zotepine versus placebo in the treatment of schizophrenic patients with stable primary negative symptoms: a randomized double-blind multicenter trial. AB - BACKGROUND: In contrast to traditional antipsychotic medication, newer, the so called atypical antipsychotic drugs promise to reduce primary negative symptoms in both, acute and chronic schizophrenia and improve quality of life. Zotepine, a D (2)5HT (2A) antagonist, was compared to placebo in this clinical trial to address these issues. METHODS: In an adaptive, 8-week multicenter, double-blind, randomized, parallel group study, efficacy and tolerability as well as influence on quality of life of zotepine and placebo were compared in 80 schizophrenic patients (53 % males, age 41 +/- 11 years) who experienced stable primary negative symptoms. Efficacy was assessed using the PANSS (primary outcome criterion: PANSS negative score), CGI, and MADRS; safety was controlled by EPS (Simpson-Angus Scale), TDRS and adverse events. Quality of life was evaluated with the SF-36 health questionnaire. RESULTS: After eight weeks of treatment with an average dose of 131 +/- 49 mg/day of zotepine, no superior efficacy of zotepine compared to placebo with respect to the PANSS subscale "negative symptoms could be demonstrated. Although patients receiving zotepine showed a pronounced reduction (per protocol set: -7.8 +/- 5.8) during treatment, it was not statistically different from a marked placebo response (-6.5 +/- 5.8). Similar small but not significant differences between zotepine and placebo were found in other subscales of the PANSS, the CGI and the MADRS. With regards to the quality of life assessment, patients under zotepine showed better results in the physical component scale and psychological well-being scale. In general, zotepine was well tolerated and there was no excess of extrapyramidal symptoms compared to placebo. CONCLUSIONS: Zotepine was not superior to placebo in reducing the severity of stable primary negative symptoms in schizophrenic patients in this trial. The main reasons for this finding were a high placebo response in a selected population, a probably too low dose of zotepine, and a short study period. However, several findings show clinical benefit of zotepine therapy in efficacy and quality of life. PMID- 15551194 TI - Olanzapine versus flupenthixol in the treatment of inpatients with schizophrenia: a randomized double-blind trial. AB - OBJECTIVE: The atypical antipsychotic olanzapine has extensively been compared with haloperidol, whereas studies vs. other (conventional) neuroleptics are scarce. This exploratory double-blind 4-week study was designed to compare the efficacy and the safety of olanzapine (OLA) and flupenthixol (FLU) which have recently been considered as a "partially atypical" antipsychotics. METHODS: Twenty-eight inpatients with schizophrenia (DSM-IV) were randomly assigned for treatment with OLA (N = 15, 5-20 mg/d) or FLU (N = 13, 5-20 mg/d). The Brief Psychiatric Rating Scale (BPRS) and the Negative Symptoms Rating Scale (NSRS), plus the Patient Global Impression (PGI) and Clinical Global Impression (CGI) scales, were used to assess the efficacy of both compounds; safety was determined by using the Simpson Angus Scale (SAS) and the Abnormal Involuntary Movement Scale (AIMS) and by assessing treatment-emergent adverse events. Non-parametric statistics were applied. RESULTS: BPRS and NSRS scores improved in both groups (exploratory tests; all p < or = 0.02). Similar results were observed for CGI Severity, CGI- and PGI-Improvement. There were no significant group differences. Responder rates (at least 40 % decrease in BPRS total) were 9/13 OLA patients (69 %) and 9/12 FLU patients (75 %). EPS events were reported only in the FLU group (p < 0.01); FLU patients needed significantly more anticholinergic medication. Weight gain was higher in OLA patients (p < 0.01). Overall, fewer patients with adverse events were observed in the OLA group (p = 0.04). No significant changes were noted on SAS and AIMS scores. CONCLUSION: Findings from this study suggest that overall and negative symptomatology improved in both treatment groups, while the safety and tolerability profiles differed for both substances. PMID- 15551195 TI - Neuroimaging and 5-HT2C receptor polymorphism: a HMPAO-SPECT study in healthy male probands using mCPP-challenge of the 5-HT2C receptor. AB - BACKGROUND: The human 5-HT (2C) receptor gene has been localized on the X chromosome and is expressed in two genetic variants. Whereas previous investigations have suggested that the 5-HT (2C) receptor gene polymorphism is critically involved in the pathogenesis of affective and eating disorders, as yet the functional consequences being associated with the rare serine variant of the 5-HT (2C) receptor in humans are unclear. METHODS: We explored by HMPAO-SPECT if a challenge with the serotonin agonist mCPP, that interacts mainly with the 5-HT (2C) receptor, provokes different patterns of regional cerebral bloodflow (rCBF) as a function of the genetic variant of the receptor. Thus we studied its action in 16 healthy male volunteers carrying the common 5-HT (2C)-cys-23 receptor gene and 16 healthy male volunteers carrying the less frequent 5-HT (2C)-ser-23 receptor gene. RESULTS: We found significant differences in rCBF between the two genotypes after mCPP infusion compared to placebo: In the cysteine group rCBF was increased in the left medial prefrontal cortex and decreased in the left anterior cingulate and right medio-temporal cortex, whereas the serine group showed an increase of rCBF in the left medio- and superior-temporal cortex and in cerebellum and a reduced rCBF in the right medial prefrontal cortex. In addition, there was a significant disordinal interaction of the genotype factors and challenge with an increase of rCBF in the serine group and a decrease in the cysteine group in the left motor cortex and calcarine cortex. Additionally, a decrease of rCBF in the serine-group and a simultaneous increase in the cysteine group was found in the right anterior and the left posterior cingulate cortex. CONCLUSION: These findings suggest that differences in the 5-HT (2C) receptor gene polymorphism has functional consequences due to a different responsiveness of the expressed 5-HT (2C) receptor variants. PMID- 15551196 TI - Modulation of P-glycoprotein function by St John's wort extract and its major constituents. AB - INTRODUCTION: Recent data suggest some relevant drug interactions caused by St John's wort extract, which can be explained by interactions with the Cytochrome P450 system or P-Glycoprotein (Pgp). Interaction with Pgp, including activation, inhibition and induction, can lead to altered plasma or brain levels of Pgp substrates. The aim of the present study was to investigate the possible interactions of St John's wort extract and most relevant constituents with the transport activity of Pgp. METHODS: We characterized the modulatory potencies in two in vitro assays using calcein-AM, first in VLB cells (a human lymphocytic leukemia cell line expressing Pgp) and second in PBCEC cells (porcine brain capillary endothelial cells). RESULTS: The extract, as well as some of the tested constituents modulate the transport by Pgp in the micromolecular range. Quercetin and hyperforin seem to be most potent. CONCLUSIONS: These findings suggest the possibility of drug interactions at the level of the gastro-intestinal absorption of drugs. Plasma levels of the constituents of St John's wort are very likely too low to interfere with Pgp at the blood-brain-barrier with the possible exception of quercetin. PMID- 15551197 TI - Open-label non-randomized versus double-blind randomized antidepressive treatment: what are the advantages of clinical decision over randomization? AB - OBJECTIVE: To study, whether and how the results from open and double-blind randomized trials on antidepressants differ. METHODS: Seventy-one patients were included in a study comparing open, non-randomized, standardized treatment with paroxetine (PAROX) and amitriptyline (AMI) after a minimum of six drug-free days (OPEN). A second group of 56 patients received the same treatment under blind randomized conditions (BLIND-RANDOM). The course of psychopathology as assessed by the Hamilton Depression Rating Scale was compared using repeated measurements ANOVA-(rm). RESULTS: While the rate of adverse events was higher in the BLIND RANDOM compared to the OPEN condition, completer-analyses revealed no differences in psychopathological outcome. CONCLUSIONS: With similar clinical outcome BLIND RANDOM trials of antidepressants may expose depressed patients to an increased risk of adverse events, when compared to OPEN conditions. However, the clinical outcome in study completers did not differ between the BLIND-RANDOM and the OPEN condition. Thus, the psychiatrist's choice may have impact on adverse events rather than on clinical outcome of antidepressant treatment. PMID- 15551198 TI - Clozapine and pregnancy--a case history. PMID- 15551199 TI - EEG abnormalities under treatment with atypical antipsychotics: effects of olanzapine and amisulpride as compared to haloperidol. PMID- 15551200 TI - Oxcarbazepine in alcohol relapse prevention--a case series. PMID- 15551201 TI - Sleepwalking associated with reboxetine in a young female patient with major depression--a case report. PMID- 15551202 TI - Simian virus 40 and human disease. PMID- 15551203 TI - Serologic evidence for exposure to simian virus 40 in North American zoo workers. AB - Some laboratories have detected DNA from the macaque polyomavirus simian virus 40 (SV40) in human tumors, but possible routes of infection remain unknown. In the present study, an enzyme immunoassay using viruslike particles (VLPs) was used to test 254 zoo workers for antibodies to SV40; 25 zoo workers with direct contact with nonhuman primates and 15 other zoo workers (23% vs. 10%, respectively; P=.01) were seropositive for SV40. Additionally, SV40 seroreactivity confirmed by competitive-inhibition experiments (i.e., blocked by addition of SV40 VLPs but not by VLPs for BK virus or JC virus, which are related human polyomaviruses) was increased in zoo workers with direct contact with nonhuman primates (10% vs. 3%, respectively; P=.04). SV40 seroreactivity therefore may reflect zoonotic exposure. PMID- 15551204 TI - Age-Specific prevalence of anal human papillomavirus infection in HIV-negative sexually active men who have sex with men: the EXPLORE study. AB - BACKGROUND: In the United States, anal cancer in men who have sex with men (MSM) is more common than cervical cancer in women. Human papillomavirus (HPV) is causally linked to the development of anal and cervical cancer. In women, cervical HPV infection peaks early and decreases after the age of 30. Little is known about the age-specific prevalence of anal HPV infection in human immunodeficiency virus (HIV)-negative MSM. METHODS: We studied the prevalence and determinants of anal HPV infection in 1218 HIV-negative MSM, 18-89 years old, who were recruited from 4 US cities. We assessed anal HPV infection status by polymerase chain reaction. RESULTS: HPV DNA was found in the anal canal of 57% of study participants. The prevalence of anal HPV infection did not change with age or geographic location. Anal HPV infection was independently associated with receptive anal intercourse (odds ratio [OR], 2.0; P<.0001) during the preceding 6 months and with >5 sex partners during the preceding 6 months (OR, 1.5; P<.0001). CONCLUSIONS: Urban, HIV-negative MSM have a stable, high prevalence of anal HPV infection across all age groups. These results differ substantially from the epidemiologic profile of cervical HPV infection in women. This may reflect differences between these populations with respect to the number of new sex partners after the age of 30 and may explain the high incidence of anal cancer in MSM. PMID- 15551205 TI - Incidence, duration, and determinants of cervical human papillomavirus infection in a cohort of Colombian women with normal cytological results. AB - Data on the incidence and determinants of human papillomavirus (HPV) infection in women >30 years old are scarce. To address this, a cohort of 1610 women--15-85 years old, HPV negative, and with normal cytological results at baseline--was monitored every 6 months for an average of 4.1 years. Information on risk factors and cervical samples for cytological testing and detection and typing of HPV DNA were obtained at each visit. The incidence of high-risk types was higher than that of low-risk types (5.0 vs. 2.0 cases/100 woman-years). The age-specific incidence curve for high-risk types was bimodal, whereas the incidence of low risk types gradually decreased with age. Infections with high-risk types lasted longer than infections with low-risk types (14.8 vs. 11.1 months). In this cohort of cytologically normal women, the incidence of cervical HPV infection was high, and the epidemiological profile of high-risk HPV types was different from that of low-risk types. PMID- 15551206 TI - Avian influenza A virus (H7N7) epidemic in The Netherlands in 2003: course of the epidemic and effectiveness of control measures. AB - An epidemic of high-pathogenicity avian influenza (HPAI) A virus subtype H7N7 occurred in The Netherlands in 2003 that affected 255 flocks and led to the culling of 30 million birds. To evaluate the effectiveness of the control measures, we quantified between-flock transmission characteristics of the virus in 2 affected areas, using the reproduction ratio Rh. The control measures markedly reduced the transmission of HPAI virus: Rh before detection of the outbreak in the first infected flock was 6.5 (95% confidence interval [CI], 3.1 9.9) in one area and 3.1 in another area, and it decreased to 1.2 (95% CI, 0.6 1.9) after detection of the first outbreak in both areas. The observation that Rh remained >1 suggests that the containment of the epidemic was probably due to the reduction in the number of susceptible flocks by complete depopulation of the infected areas rather than to the reduction of the transmission by the other control measures. PMID- 15551207 TI - Evaluation of combined live, attenuated respiratory syncytial virus and parainfluenza 3 virus vaccines in infants and young children. AB - We evaluated a combination respiratory syncytial virus (RSV) and parainfluenza 3 virus (PIV3) live, attenuated intranasal vaccine for safety, viral replication, and immunogenicity in doubly seronegative children 6-18 months old. RSV cpts 248/404 and PIV3-cp45 vaccines were combined in a dose of 10(5) plaque-forming units of each per 0.5-mL dose and compared with monovalent vaccines or placebo. The virus shedding pattern of RSV was not different between monovalent RSV cpts 248/404 vaccine and combination vaccine. Modest reductions in the shedding of PIV3-cp45 vaccine virus were found after the administration of RSV cpts-248/404 and PIV3-cp45 vaccine, relative to monovalent PIV3 vaccine; 16 (76%) of 21 children given combination vaccine shed PIV3-cp45 versus 11 (92%) of 12 of those given monovalent PIV3 vaccine. Both vaccines were immunogenic, and antibody responses were similar between the monovalent groups and the combination group. Combined RSV/PV3 vaccine is feasible for simultaneous administration, and further studies are warranted. PMID- 15551208 TI - Induction of sterilizing immunity against West Nile Virus (WNV), by immunization with WNV-like particles produced in insect cells. AB - No specific vaccine for West Nile virus (WNV) is currently available for human use. In the present study, we describe the generation of WNV-like particles (WNV LPs) in insect cells by use of recombinant baculoviruses expressing the WNV structural proteins prME or CprME. BALB/c mice immunized with purified WNV-LPs developed WNV-specific antibodies that had potent neutralizing activities. Mice immunized with prME-like particles (prME-LPs) showed no morbidity or mortality after challenge with WNV. Immunization with prME-LPs can induce sterilizing immunity without producing any evidence of viremia or viral RNA in the spleen or brain. These results suggest that WNV-LPs hold promise as a vaccine candidate for WNV infection. PMID- 15551209 TI - Detection of novel organisms associated with salpingitis, by use of 16S rDNA polymerase chain reaction. AB - Although Chlamydia trachomatis and Neisseria gonorrhoeae are established causes of salpingitis, the majority of cases have no known etiology. We used broad-range 16S rDNA polymerase chain reaction to identify novel, possibly uncultivable, bacteria associated with salpingitis and identified bacterial 16S sequences in Fallopian-tube specimens from 11 (24%) of 45 consecutive women with laparoscopically confirmed acute salpingitis (the case patients) and from 0 of 44 women seeking tubal ligations (the control subjects) at Kenyatta National Hospital, Nairobi, Kenya. Bacterial phylotypes most closely related to Leptotrichia spp. were detected as the sole phylotypes in 1, and mixed with other bacterial phylotypes in 2, specimens. Novel bacterial phylotypes and those associated with bacterial vaginosis, including Atopobium vaginae, were identified in 3 specimens. N. gonorrhoeae and Streptococcus pyogenes were identified in 2 and 1 specimens, respectively. The finding of novel phylotypes associated with salpingitis has important implications for the etiology, pathogenesis, and treatment of this important reproductive-tract disease syndrome. PMID- 15551210 TI - Virulence factor profiles and phylogenetic background of Escherichia coli isolates from veterans with bacteremia and uninfected control subjects. AB - BACKGROUND: Escherichia coli is the most common cause of gram-negative bloodstream infections, causing an estimated 40,000 deaths from sepsis each year in the United States. The present study sought to determine specifically which virulence factors (VFs) and phylogenetic groups of E. coli are epidemiologically associated with bacteremia. METHODS: E. coli isolates from 63 veterans with bacteremia and rectal isolates from 71 matched uninfected control subjects were compared both for phylogenetic group and for the presence of VFs and O antigens. RESULTS: Bacteremia isolates exhibited a significantly greater prevalence of most VFs studied. In multivariate logistic regression analysis, ompT (outer membrane protein T) was the strongest VF predictor of bacteremia (P<.001). Despite the concentration of most individual VFs within group B2, bacteremia and rectal isolates differed little by phylogenetic distribution, a finding explained by the greater prevalence of VFs among bacteremia isolates than rectal isolates within groups B2 and D. CONCLUSIONS: Although phylogenetic group partially corresponds with virulence potential in E. coli bacteremia, VFs are more-powerful predictors of pathogenic potential. Bacteremia isolates exhibit an arsenal of VFs that distinguishes them from rectal isolates from uninfected hosts, which makes these differences attractive potential targets in vaccine or drug development. PMID- 15551211 TI - An interventional approach to block brain damage caused by Shiga toxin-producing Escherichia coli infection, by use of a combination of phosphodiesterase inhibitors. AB - We tested the combination of phosphodiesterase (PDE) 3 and PDE4 inhibitors as an interventional approach to prevent the development of brain damage after Shiga toxin (Stx)-producing Escherichia coli (STEC) infection, using mice with protein calorie malnutrition. The combination consisted of pentoxifylline and rolipram; the dose of each inhibitor was 7.5 mg/kg. Treatment with this combination, which was administered intraperitoneally twice daily at 12-h intervals, increased serum concentrations of each inhibitor to >2 microg/mL and afforded significant levels of protection when it was continued for 3 days, starting on day 2 (95% survival rate; P<.001) or day 3 (63% survival rate; P<.01) of infection. The treatment reduced plasma levels of Stx2; consequently, immunoreactions of Stx2 were not found in the brain, and survivors did not show neurologic symptoms. Protection was associated with decreased levels of tumor necrosis factor (TNF)- alpha and increased production of interleukin-10 in serum, the brain, and the cecum. Although the combination at doses >2 microg/mL reduced Gb3 content of and Stx2 binding to Caco-2 cells, its ability to suppress production of TNF- alpha seemed to be more important for the decrease in cell-bound Stx2 in intestinal epithelial cells. Therefore, the combination of PDE3 and PDE4 inhibitors might be used as an interventional approach to prevent brain damage caused by STEC infection. PMID- 15551212 TI - Mycobacterial dissemination and cellular responses after 1-lobe restricted tuberculosis infection of genetically susceptible and resistant mice. AB - BACKGROUND AND METHODS: To study mycobacterial dissemination and immune-cell trafficking in tuberculosis, we developed a mouse model in which we introduced 1 microL of Mycobacterium tuberculosis directly into the middle lobe of the right lung. We investigated the kinetics of both mycobacterial spread to different anatomical sites and recruitment of phagocytes and activated lymphocytes. RESULTS: Mycobacterial dissemination was independent of susceptibility to infection and was identical in H-2-congenic mouse strains with high and low resistance to tuberculosis. In resistant mice, recruitment of phagocytic cells to the uninfected lung occurred before the appearance of mycobacteria and decreased shortly thereafter. In susceptible mice, this recruitment was delayed in both lungs but increased during a 10-week period. Recruitment of CD4+ and CD8+ lymphocytes to the contralateral lung was observed before mycobacterial dissemination in both strains, so mycobacterial seeding of secondary tissues occurred in the presence of immune lymphocytes. In resistant mice, more T cells expressed the CD44hi CD62lo activation phenotype, and higher levels of interferon gamma were produced. CONCLUSIONS: Mycobacterial spread to lymphoid organs preceded spread to the initially uninfected contralateral lung. Genetic differences in susceptibility to tuberculosis are associated with differences in dynamics of the immune response, rather than differences in mycobacterial trafficking. PMID- 15551214 TI - Emergence of penicillin-nonsusceptible Streptococcus pneumoniae clones expressing serotypes not present in the antipneumococcal conjugate vaccine. AB - BACKGROUND: Penicillin-nonsusceptible Streptococcus pneumoniae isolates are confined mainly to a few serogroups. Capsular transformation may serve as a mechanism for spreading antibiotic resistance to new serotypes. METHODS: Antibiogram and molecular typing, by pulsed-field gel electrophoresis (PFGE), were performed on 46 nasopharyngeal and middle ear fluid (MEF) isolates expressing serotype 11A, 45 MEF isolates expressing serotype 15B/C (recovered during 1998-2003 from Israeli children <5 years old), and 57 MEF isolates expressing serotype 19F (recovered during 1998-2001 from Costa Rican children <7.5 years old). RESULTS: PFGE patterns showed that 49 (86%) of 57 serotype 19F isolates and 19 (41%) of 46 serotype 15B/C isolates were closely related. The vast majority of these isolates (80% of serotype 19F and 100% of serotype 15B/C isolates) were nonsusceptible to penicillin. Multilocus sequence typing (MLST) data show that the serotype 15B/C isolates belonged to the ST346 cluster, whereas the serotype 19F isolates were a single-locus variant of ST346. For serotype 11A isolates, PFGE patterns and MLST analysis showed that 8 (80%) of the 10 penicillin-nonsusceptible isolates belonged to a single clone--namely, ST156- which was identical to the international Spain9V-3 clone. CONCLUSIONS: Penicillin nonsusceptible pneumococcal clones of serotypes not related to those included in the 11-valent conjugate vaccines may derive from capsular transformation of vaccine-related serotypes. Of particular concern was the detection of serotype 11A variants of the successful international Spain9V-3 clone. This phenomenon, although seemingly rare at present, can have implications for the long-term effectiveness of the conjugate vaccines. PMID- 15551213 TI - Comparative analysis of different vaccine constructs expressing defined antigens from Mycobacterium tuberculosis. AB - BACKGROUND: Studies of different vaccine constructs have demonstrated variable efficacy against Mycobacterium tuberculosis in animal models. Despite the fact that these vaccines have used one or another of a very small number of immunodominant antigens, a direct comparison of the relative efficacy of the antigens and delivery systems has been difficult, because the studies have used different parameters for assessment. METHODS: We compared the efficacies of the most commonly used vaccine constructs--adjuvanted protein, plasmid DNA, and live bacterial vectors--bearing the immunodominant secreted antigens early secreted antigen target-6 and antigen 85B, either alone or as a fusion protein. Mice were vaccinated with these constructs, and the effects of different delivery systems on protective efficacy (as assessed by survival studies and by monitoring bacterial load) and antigen-specific responses (including the contribution of CD4 and CD8 T cells to these responses) were assayed by various methods. RESULTS: The relative efficacy of different vaccines is dependent on the delivery system, the antigen, and the animal model. Likewise, the relative immunodominance of individual antigens in the fusion molecule is altered by the choice of delivery system. CONCLUSION: These results clearly demonstrate the importance of assessing vaccine function by use of multiple parameters and indicate which parameters are most reliable for assessing vaccine efficacy. PMID- 15551215 TI - In vitro antienterococcal activity explains associations between exposures to antimicrobial agents and risk of colonization by multiresistant enterococci. AB - We compared ceftriaxone and piperacillin-tazobactam at doses ranging from 0.1 to 2 times the human equivalent daily dose (HEDD), to determine their impact on gastrointestinal colonization by ampicillin- and vancomycin-resistant Enterococcus faecium C68 in a mouse model. Ceftriaxone failed to promote colonization at doses up to 0.25 times the HEDD, whereas piperacillin-tazobactam promoted colonization at doses up to 0.5 times the HEDD. Ceftriaxone promoted colonization at doses at least 0.5 times the HEDD, whereas piperacillin tazobactam inhibited colonization at doses at least 0.75 times the HEDD. Both piperacillin-tazobactam and ceftriaxone inhibited colonization by an enterococcal strain devoid of low-affinity penicillin-binding protein-5 (significantly increasing its susceptibility to these agents), at doses that promoted colonization by E. faecium C68. These results support a model in which the impact that different beta -lactam agents have on colonization by VRE is related to the level of the beta -lactam agent's intrinsic antienterococcal activity against the colonizing strain. PMID- 15551217 TI - Why does HIV infection not lead to disseminated strongyloidiasis? AB - We investigated the hypothesis that host immunosuppression due to advancing human immunodeficiency virus (HIV) disease favors the direct development of infective larvae of Strongyloides stercoralis, which may facilitate hyperinfection and, hence, disseminated strongyloidiasis. To do this, we sought correlations between the immune status of the subjects and the development of S. stercoralis infections. Among 35 adults, there were significant negative rank correlations between CD4+ cell counts and the proportions of free-living male and female worms. Thus, in individuals with preserved immune function, direct development of S. stercoralis is favored, whereas, in individuals with lesser immune function, indirect development is relatively more common. These results may explain the notable absence of disseminated strongyloidiasis in advanced HIV disease. Because disseminated infection requires the direct development of infective larvae in the gut, the observed favoring of indirect development in individuals immunosuppressed by advancing HIV disease is not consistent with the promotion of disseminated infection. PMID- 15551216 TI - Pharmacokinetics and safety of stavudine in HIV-infected pregnant women and their infants: Pediatric AIDS Clinical Trials Group protocol 332. AB - This study evaluates the safety, tolerance, and pharmacokinetics of stavudine (d4T) in human immunodeficiency virus (HIV)-infected zidovudine (ZDV) intolerant/refusing pregnant women and of single-dose d4T in their infants. Women received d4T and lamivudine (3TC) from enrollment until labor. During labor, women received oral 3TC and either intravenous or oral d4T. Infants received ZDV and 3TC for 6 weeks and a single dose of oral d4T at weeks 1 and 6. Mean maternal antenatal d4T pharmacokinetics (terminal plasma half-life [T1/2], 83.5+/-16.8 min; area under the plasma-concentration time curve [AUC0-infinity), 81.6+/-22.0 microg.min/mL; n=6) were not significantly different from those during labor (T(1/2), 87.3+/-24.7 min; AUC0-infinity, 88.1+/-16.6 microg.min/mL; n=6). Umbilical-cord and maternal plasma concentrations were not significantly different from one another. The oral clearance of d4T in infants was significantly greater at week 6 versus week 1 (6.8+/-1.0 vs. 5.6+/-1.2 mL/min/kg). There were no toxicities, in women or infants, that required discontinuation or modification of the study drug. No infants had positive HIV viral diagnostic tests. d4T with or without 3TC is a potential alternative to ZDV for HIV-infected pregnant women. PMID- 15551218 TI - Longitudinal analysis of nef/long terminal repeat-deleted HIV-1 in blood and cerebrospinal fluid of a long-term survivor who developed HIV-associated dementia. AB - We studied the evolution and compartmentalization of nef/long terminal repeat (nef/LTR)-deleted human immunodeficiency virus type 1 (HIV-1) from a long-term survivor who developed HIV-associated dementia (HIVD). Analysis of sequential blood-derived HIV-1 isolated before and during HIVD revealed a persistent R5X4 phenotype and a progressive loss of nef/LTR sequence; in contrast, HIV-1 present in cerebrospinal fluid during HIVD had an R5 phenotype, distinct nef/LTR sequence of unique deletions and additional nuclear factor- kappa B sites and specificity factor-1 sites, and enhanced transcriptional activity, compared with the blood derived isolates. Thus, nef/LTR-deleted HIV-1 strains may undergo compartmentalized evolution in long-term survivors and cause neurologic disease. PMID- 15551219 TI - Wasting syndrome and disruption of the somatotropic axis in simian immunodeficiency virus-infected macaques with Mycobacterium avium complex infection. AB - Mycobacterium avium is a common opportunistic infection of patients with acquired immunodeficiency syndrome (AIDS). We used the simian immunodeficiency virus (SIV) infected rhesus macaque (Macaca mulatta) model to examine whether disseminated M. avium is associated with disruption of the somatotropic axis in AIDS. Macaques were followed prospectively, and body composition was determined by dual-energy x ray absorption. Serum levels of insulin-like growth factor (IGF)-1, IGF binding protein-3, growth hormone (GH), and somatostatin were measured. SIV-infected macaques inoculated with mycobacteria had significant changes in body composition, perturbations of the somatotropic axis (characterized by increased GH/IGF-1 ratios) (day 0 [2.21] vs. day of death [DOD] [28.06]; P=.015, Mann Whitney rank sum test), and increased serum somatostatin levels (day 0 [2.00 ng/mL] vs. DOD [8.58 ng/mL]; P=.026, Mann-Whitney rank sum test). These data document alterations in the somatotropic axis secondary to experimental disseminated M. avium infection and suggest that similar changes may contribute to alterations in body composition during AIDS. PMID- 15551220 TI - Elimination programs: monitoring the effectiveness of surveillance. PMID- 15551223 TI - Veterinary medical education in Iraq. AB - Iraq is an agricultural country with a large population of animals: sheep, goats, cattle, water buffaloes, horses, donkeys, mules, and camels. In the 1980s, the successful poultry industry managed to produce enough table eggs and meat to satisfy the needs of the entire population; at one time, the thriving fish industry produced different types of fish for Iraqis' yearly fish consumption. There are four veterinary colleges in Iraq, which have been destroyed along with the veterinary services infrastructure. Understandably, improvements to the quality of veterinary education and services in Iraq will be reflected in a healthy and productive animal industry, better food quality and quantity, fewer zoonotic diseases, and more income-generating activities in rural areas. Thus, if undergraduate, graduate, and continuing education programs are improved, the veterinary medical profession will attract more competent students. This will satisfy the country's increased demand for competent veterinarians in both public and private sectors. Although Iraq has an estimated 5,000-7,000 veterinarians, there is a need for quality veterinary services and for more veterinarians. In addition, there is a need for the improvement of veterinary diagnostic facilities, as zoonotic diseases are always highly probable in this region. This article provides insight into the status of veterinary medical education and veterinary services in Iraq before and after the 1991 Gulf War and gives suggestions for improvement and implementation of new programs. Suggestions are also offered for improving veterinary diagnostic facilities and the quality of veterinary services. Improving diagnostic facilities and the quality of veterinary services will enhance animal health and production in Iraq and will also decrease the likelihood of disease transmission to and from Iraq. Threats of disease transmission and introduction into the country have been observed and reported by several international organizations. PMID- 15551224 TI - Bourgelat's vision for veterinary education and the remarkable spread of the veterinary "meme". PMID- 15551222 TI - Association between maternal age and meiotic recombination for trisomy 21. AB - Altered genetic recombination has been identified as the first molecular correlate of chromosome nondisjunction in both humans and model organisms. Little evidence has emerged to link maternal age--long recognized as the primary risk factor for nondisjunction--with altered recombination, although some studies have provided hints of such a relationship. To determine whether an association does exist, chromosome 21 recombination patterns were examined in 400 trisomy 21 cases of maternal meiosis I origin, grouped by maternal age. These recombination patterns were used to predict the chromosome 21 exchange patterns established during meiosis I. There was no statistically significant association between age and overall rate of exchange. The placement of meiotic exchange, however, differed significantly among the age groups. Susceptible patterns (pericentromeric and telomeric exchanges) accounted for 34% of all exchanges among the youngest class of women but only 10% of those among the oldest class. The pattern of exchanges among the oldest age group mimicked the pattern observed among normally disjoining chromosomes 21. These results suggest that the greatest risk factor for nondisjunction among younger women is the presence of a susceptible exchange pattern. We hypothesize that environmental and age-related insults accumulate in the ovary as a woman ages, leading to malsegregation of oocytes with stable exchange patterns. It is this risk, due to recombination independent factors, that would be most influenced by increasing age, leading to the observed maternal age effect. PMID- 15551225 TI - Editorial: food animal veterinary futures. PMID- 15551226 TI - Veterinary medicine and the lifeboat test: a perspective on the social relevance of the veterinary profession in the 21st century. PMID- 15551227 TI - Emerging challenges in public health protection, food safety and security: veterinary needs in the USDA's Food Safety and Inspection Service. AB - Meeting the needs of public service practice is a responsibility of the veterinary profession. The United States Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) has undergone significant change since 1996, when the final rule on Pathogen Reduction and Hazard Analysis and Critical Control Point (HACCP) Systems and its regulations were published in response to food-borne illnesses and deaths due to E. coli 0157:H7 in undercooked hamburgers. As a result, the role of the veterinarian is changing from a focus on carcass inspection (reactive) to scientific-based systems analysis and enforcement (preventive). With a large pool of veterinarians eligible to retire, a critical shortage of field veterinarians is predicted. The purpose of this article is to raise educators' awareness of this need, of the competencies required, and of the challenges and opportunities for veterinarians in the new public health-focused FSIS. An invitation to collaborate with the agency is offered to help meet emerging workforce requirements in public health practice. PMID- 15551228 TI - Food animal veterinary medicine: leading a changing profession. AB - The veterinary profession has gone through periods of profound change in response to economic and social changes. We are currently in another such period: profound change is required in order for the profession to remain relevant in a marketplace where a rapidly expanding knowledge base and new technologies demand an ever-increasing level of expertise in a greater variety of areas. However, the veterinary profession is perceived both internally and by the public as possessing a narrow set of skills that supports a narrow group of careers focused on salvaging ill or injured companion animals. It will be necessary to dramatically change the way veterinary students are recruited and trained, as well as how graduate veterinarians are licensed and provided continuing education, in order for the veterinary profession to capitalize on our historical strengths and provide service and leadership in a greater diversity of career paths. Even though the number of veterinarians needed to provide primary care for livestock is decreasing, both the level of expertise demanded by livestock owners and the value of veterinary involvement on livestock farms are increasing. Colleges of veterinary medicine appear challenged to meet the changing needs for veterinary services in animal agriculture because of the declining percentage of veterinary students interested in food animal careers. Fortunately for animal agriculture, the skill set needed by food animal veterinarians is also needed by several emerging segments of the veterinary profession that have tremendous potential for rapid growth, including employment in all segments of food production systems, environmental monitoring and management, bio-security and disease eradication, laboratory diagnostics, and federal regulatory and bio defense roles. Like previous periods of profound change, this moment in history will require creative thought, open discussion, and a willingness to step into the unknown. PMID- 15551229 TI - Practitioner-defined competencies required of new veterinary graduates in food animal practice. PMID- 15551230 TI - The UC Davis Early Dairy Experience Program. PMID- 15551231 TI - Minnesota's Transition Management Facility: a private--public partnership in dairy veterinary education and applied research. PMID- 15551232 TI - A coordinated teaching program for future dairy practitioners at the university of Wisconsin-Madison, School of Veterinary Medicine. AB - The University of Wisconsin School of Veterinary Medicine (UW-SVM) has implemented a variety of strategies to optimize teaching in dairy herd medicine. These include the provision of opportunities for dairy cow handling and management using a dairy teaching herd for veterinary students throughout the four-year curriculum, exposure for all students in their final year to a substantial first-opinion dairy case load using a private practice-based ambulatory clinic rotation, and, finally, the teaching of dairy herd health management and problem solving in a group of four final-year elective production medicine clinical rotations. On average, since 1986, 32.6% of each graduating class attended at least one elective production medicine rotation, with a range from 19.0% to 43.4%. For those University of Wisconsin students who could be traced, 65% were still actively involved in some aspect of dairy practice, representing a range of between seven and 17 students per year since the start of the program. The advantages and disadvantages of operating a "regional center of excellence" for training students from out-of-state institutions are discussed. PMID- 15551233 TI - Use of a non-traditional university ambulatory practice to teach large animal medicine. AB - While many other veterinary schools have moved away from a traditional university based ambulatory practice, the Ohio State University's Large Animal Practice has continued to provide a cost-effective and valuable method of preparing students for today's careers in veterinary medicine. The practice provides a full array of services to production, equine, and camelid clients, including herd health, individual animal medicine and surgery, and emergency services. Acquiring established practices from alumni has formed the client base. Four full-time veterinarians operate the clinic. While these same clinicians do some classroom teaching, their primary responsibility is devoted to the five to six fourth-year veterinary students who rotate through the clinic every two weeks. Teaching methods and objectives for these students include case discussions, homework, truck quiz books, and practice management issues. Financially, the clinic runs as a private practice, with minimal support from the college (201,000 US dollars per fiscal year) and a gross income of 676,000 US dollars per year. Thus, in a cost effective manner, this required core ambulatory rotation provides students with a scientific learning experience that exposes them to all aspects of large animal production medicine in a real-world setting. PMID- 15551234 TI - The Opportunities Map at Cornell University: finding direction in dairy production medicine. AB - Discussion between faculty and interested students revealed the existence of a multitude of opportunities in dairy production medicine at the College of Veterinary Medicine at Cornell University. Many of these were not well known to students, or even to some of the faculty, and the means of accessing specific learning experiences were sometimes obscure. Together, an informal group of faculty, students, and alumni set about cataloging available educational opportunities, resulting in a 31-page publication referred to as the "Opportunities Map." Essentially a student handbook for production medicine students, the Opportunities Map at Cornell helps guide the travel of food animal interested students through the curriculum without missing the important highlights along the way. The map was originally developed to chronicle the opportunities and resources available to students, but it has also been used to foster face-to-face communications between students and faculty, to welcome incoming students with production animal interests, and to provide a baseline description for further discussion about the curriculum. PMID- 15551235 TI - Lessons learned: what have international experiences in large livestock disease outbreaks taught us? Some personal insights. PMID- 15551236 TI - Food supply veterinary medicine: creating an awareness of livestock security risks. PMID- 15551238 TI - Diversity: a professional imperative. PMID- 15551239 TI - Diversity and changing demographics: how they will affect veterinary medicine. PMID- 15551240 TI - A historical overview of African American veterinarians in the United States, 1889- 2000. AB - The annals of veterinary medical history rarely mention the presence of African American veterinarians and other minorities. Between 1889 and 1948, records show, a meager 70 African Americans graduated from veterinary schools in the United States and Canada. It was not until the veterinary school at Tuskegee (Institute) University was established in 1945 that a significant increase in the number of African American veterinarians occurred in the United States, and over the ensuing years their participation in every facet of the profession has been striking. Their employment in various areas of the profession and their successful performance in the workforce have done much to dispel stereotypical perceptions about minorities. Despite demographic data indicating that the United States is moving rapidly toward a multicultural society, recruitment programs to increase the number of African American students and faculty at the 27 US veterinary colleges have not kept pace with the declared goals of ethnic diversity. If the needs of a changing culture are to be met, veterinary medical education must look toward more ethnic inclusion in the student body and faculty. To that end, the Iverson C. Bell Symposium has consistently advocated the adoption of new and creative methods for increasing minority student enrollment and expanding faculty opportunities in the nation's veterinary colleges. PMID- 15551241 TI - Reasons for the lack of racial diversity in veterinary medicine. PMID- 15551242 TI - Veterinary medical colleges' diversity awareness. PMID- 15551243 TI - Affirmative action in college admissions: the case for diversity. PMID- 15551244 TI - Skills, knowledge, aptitudes, and attitudes colloquium. AB - Three projects recently funded by the American Veterinary Medical Association (AVMA) through the National Commission on Veterinary Economic Issues (NCVEI) focused on the veterinary school applicant pool, leadership skills in the veterinary profession, and a veterinary teaching hospital business model, respectively. The Skills, Knowledge, Aptitude, and Attitude (SKAs) Colloquium was designed to present the results of these three projects, to discuss their importance for the future of the veterinary profession, and to develop action plans accordingly. In all, 24 veterinary colleges were represented at the workshop and a total of 72 attendees participated, achieving a broad representation of the veterinary profession ( both academic and non-academic). Through an orchestrated combination of general sessions and facilitated small group discussions, recommendations for implementation and initial action plans for next steps were developed. From these, a list of potential AAVMC follow-up activities was developed, including advocating and facilitating programs across schools to engage and educate faculty regarding the results of these projects; developing realistic information on careers in veterinary medicine; organizing an AAVMC leadership consortium; working toward further development and implementation of the veterinary teaching hospital (VTH) business model; coordinating and sponsoring a national forum on the future of the VTH; reviewing admissions processes; integrating leadership into veterinary curricula; and organizing opportunities for faculty development in leadership. PMID- 15551245 TI - Massive pneumoperitoneum and pneumomediastinum with subcutaneous emphysema after endoscopic sphincterotomy. PMID- 15551246 TI - Use of herbal supplements for chronic liver disease. AB - BACKGROUND & AIMS: Complementary and alternative medicine (CAM) is becoming popular among patients with liver disease. Although there is a growing body of evidence regarding potential mechanisms of action of these and other herbs, caution must be used to interpret the results of the few clinical trials available. Our goal was to discuss the biologic rationale for the use of specific herbs (silymarin, glycyrrhizin, sho-saiko-to, Phyllanthus amarus , Picrorrhiza kurroa , Compound 861, CH-100, and LIV.52) in the treatment of chronic liver diseases, as well as the evidence for their efficacy and adverse effects according to clinical trials. METHODS: Because of the relative paucity of clinical studies using herbs, every trial published in English was reviewed. RESULTS: Although many trials suggest that these herbs can decrease serum transaminase levels, the effects on hepatic histopathology and long-term survival are either poorly studied or conflicting. LIV.52 has been withdrawn from the market because of deleterious effects in patients with liver disease. CONCLUSIONS: Based on current evidence, we cannot recommend the use of herbal supplements for the routine treatment of any chronic liver disease and further well-designed clinical trials are necessary. PMID- 15551247 TI - Complementary and alternative medicine in gastroenterology: the good, the bad, and the ugly. AB - A large proportion of the American population avails itself of a variety of complementary and alternative medicine (CAM) interventions. Allopathic practitioners often dismiss CAM because of distrust or a belief that there is no sound scientific evidence that has established its utility. However, although not widely appreciated, there are thousands of randomized controlled trials (RCTs) that have addressed the efficacy of CAM. We reviewed the RCTs of herbal and other natural products, acupuncture, and homeopathy as examples of typical CAM modalities, focusing on conditions of interest to gastroenterologists. Peppermint (alone or in combination) has supportive evidence for use in patients with dyspepsia, irritable bowel syndrome, and as an intraluminal spasmolytic agent during barium enemas or endoscopy. Ginger appeared to be effective in relieving nausea and vomiting due to motion sickness or pregnancy. Probiotics were useful in childhood diarrhea or in diarrhea due to antibiotics; one particular formulation (VSL#3) prevented pouchitis. Acupuncture appeared to ameliorate postoperative nausea and vomiting and might be useful elsewhere. There is even a suggestion that homeopathy has efficacy in treatment of gastrointestinal problems or symptoms. The major problem in interpreting these CAM data is the generally low quality of the RCTs, although that quality might not be different compared to RCTs in the general medical literature. Gastroenterologists should become familiar with these techniques; it is likely that their patients already are. PMID- 15551248 TI - Complications of endoscopic biliary sphincterotomy: the first cut is the deepest. PMID- 15551249 TI - Prospective evaluation of pancreatic sphincterotomy as a precut technique for biliary cannulation. AB - BACKGROUND & AIMS: Pancreatic sphincterotomy has been described as an effective technique to obtain biliary access after standard methods fail. This prospective study evaluates its efficacy and compares its complication rate to conventional biliary sphincterotomy (BS). METHODS: Between January 2001 and January 2004, patients in whom biliary cannulation failed underwent a pancreatic precut sphincterotomy (PPS) and were analyzed prospectively. Multivariate analysis was performed on the following variables with regard to their ability to predict successful biliary cannulation: age, gender, time to access bile duct after precut (< or =10 or >10 minutes), final diagnosis, and operator. Complications of PPS were then compared with those resulting from endoscopic retrograde cholangiopancreatography with BS in 120 patients examined during the same period of time and matched by sex, gender, and disease process. RESULTS: One hundred sixteen patients (50 male), mean age 58 +/- 16 years, underwent PPS. Immediate biliary access was achieved after pancreatic precut in 99 cases (85%). Complications occurred in 14 patients (12%): 3 (2.6%) postsphincterotomy bleeding, 9 (8%) pancreatitis (8 mild, 1 moderate), and 2 (1.7%) retroperitoneal perforation managed conservatively. The factor statistically associated with successful biliary cannulation was the amount of elapsed time between completing the PPS and obtaining biliary access. No statistical difference was identified in the complication rate of pancreatitis between the PPS and BS groups. CONCLUSIONS: PPS is an effective precut technique to facilitate biliary cannulation. Success is correlated to the speed of biliary access after precut. In expert hands, its rate of pancreatitis is similar to endoscopic retrograde cholangiopancreatography with BS. PMID- 15551250 TI - Outcomes of surgical fundoplication in children. AB - BACKGROUND & AIMS: Surgical fundoplication has been recommended for children with persistent GERD. The purpose of this study was to determine the frequency of postoperative symptoms requiring medical evaluation and/or treatment after fundoplication in children with or without associated medical disorders. METHODS: In a retrospective cohort study, we reviewed the medical records of all children who underwent fundoplication during 1996-1999. Data were collected to analyze the following: (1) postoperative complications, (2) postoperative symptoms, (3) procedures performed to evaluate postoperative symptoms, (4) medical treatment, and (5) repeat surgery. RESULTS: A total of 198 children underwent fundoplication, and 176 (89%) came for follow-up evaluation within 2 months after surgery. The median age at the time of surgery was 2.1 y (range, 6 mo-18 y) and the median duration of follow-up was 2.0 y (range, 1.2-4.8 y). A total of 130 (74%) children had one or more associated medical disorders including neurodevelopmental delay (70%), cystic fibrosis (8%), tracheoesophageal anomalies (8%), bronchopulmonary dysplasia (8%), and reactive airway disease (35%). Postoperatively, children with associated medical disorders had a higher frequency of lung infections (52% vs. 22%, P = .03) and dumping syndrome (2% vs. 0%, P = .05). Most children (63%) received evaluation and treatment for symptoms suggestive of recurrent reflux despite fundoplication. CONCLUSIONS: Nearly two thirds of children who received fundoplication and were followed-up within 2 months after surgery either have symptoms or receive medical therapy for reflux. Fundoplication for the control of GERD in children needs further evaluation. PMID- 15551251 TI - A community-based, controlled study of the epidemiology and pathophysiology of dyspepsia. AB - BACKGROUND & AIMS: Dyspepsia is common in clinical practice and in the community. The relationship of the symptoms to meals and the pathophysiology in community dyspeptic patients is unclear. The purpose of this study was to measure symptoms, demographic features, and gastric motor and sensory functions associated with dyspepsia in the community. METHODS: A Modified Bowel Disease Questionnaire was mailed to a random sample of Olmsted County, MN, residents. Dyspeptic patients and healthy controls identified among community respondents completed further questionnaires, Helicobacter pylori serology, gastric emptying by scintigraphy, gastric accommodation by 99mTc-single-photon emission computed tomography imaging, and postprandial symptoms and satiation by a nutrient drink test. RESULTS: A total of 34.1% of community respondents reported dyspepsia within the past year, frequent (at least 25% of the time in the past year) in 17.5%, and 18.4% reported meal-related dyspepsia. Dyspepsia was frequent and related to meals in 10.8% of respondents. Compared with nondyspeptic controls, community dyspepsia was associated with higher aggregate symptom scores and bloating after a fully satiating meal. Community dyspepsia also was associated with higher somatization scores (P = .001), reporting of other somatic symptoms (P = .07), and general severity score on the symptom checklist 90 (P = .01), but not with disordered motor or sensory function. Gastric volumes, gastric emptying, and maximum tolerated volumes were not significantly different between community controls and dyspeptic patients. CONCLUSIONS: Meal-related dyspepsia is an important component of dyspepsia in the community. Community dyspeptic patients have higher symptom scores after a fully satiating meal, consistent with gastric hypersensitivity. This is associated with higher somatization scores rather than disorders of gastric emptying or volumes. PMID- 15551252 TI - A 10-day levofloxacin-based therapy in patients with resistant Helicobacter pylori infection: a controlled trial. AB - BACKGROUND & AIMS: Antibiotic resistance is a major issue in anti- Helicobacter pylori treatment. This study was aimed at assessing the efficacy of 2 therapies in patients with resistant H pylori infection. METHODS: Patients who had failed 1 or more eradication regimens underwent upper gastrointestinal endoscopy and 2 antral and 2 corpus biopsy specimens were taken for histology and culture. Metronidazole, clarithromycin, and amoxicillin resistance were determined by E test. Patients were randomly assigned to 2 therapies: 1 group received pantoprazole 40 mg, amoxicillin 1 g, levofloxacin 250 mg, all twice daily for 10 days, and the other group was treated with omeprazole 20 mg twice daily for the first week and omeprazole 20 mg twice daily, tetracycline 250 mg 4 times daily, metronidazole 500 mg twice daily, and bismuth subcitrate 240 mg twice daily for the second week. Therapeutic success was evaluated by 13C urea breath test after 4 weeks of treatment. RESULTS: We enrolled 44 patients in the levofloxacin-based regimen and 46 patients in the quadruple therapy. The former was successful in 31 of 44 (70%; 95% confidence interval: 53-87) and the latter in 17 of 46 (37%; 95% confidence interval: 23-47) patients, using intention-to-treat (ITT) analysis (P < .001). The rates of H pylori resistance to metronidazole, clarithromycin, and amoxicillin were 46%, 12%, and 0%, respectively. Resistance to both metronidazole and clarithromycin was found in 10% of cases. CONCLUSIONS: Triple therapy containing levofloxacin was better than quadruple therapy. The 70% success rate observed indicates that 10 days of pantoprazole, amoxicillin, and levofloxacin should be considered in patients who had failed 1 or more eradication regimens. PMID- 15551253 TI - CARD15 gene mutations and risk for early surgery in pediatric-onset Crohn's disease. AB - BACKGROUND & AIMS: The risk for Crohn's disease (CD) is determined in part by genetic factors. Three recently described mutations in the CARD15(NOD2) gene have been associated with adult-onset CD. We investigated the effect of CARD15 mutations on disease manifestation, disease progression, and the risk for early surgery in childhood-onset CD. METHODS: Genotyping for 3 CARD15 mutations: R702W, G908R, and 3020insC, was performed in 186 children with CD from a prospective cohort. A transmission-disequilibrium test was used to test for association with CD. Genotype with disease location and behavior was tested with logistic regression analysis. The effect of mutations on surgical outcome was evaluated using a Cox proportional hazard analysis. RESULTS: The mean age at CD diagnosis was 12.4 years. The frequency of allelic mutations observed was 6.6% for R702W, 6% for G908R, and 13.1% for 3020insC. Of Caucasian CD children, 42% had at least one CARD15 mutation. None of the non-Caucasian children with CD had any CARD15 mutation. A significant association was detected for 3020insC (P = .0045). Ileal location (odds ratio, 4.3; P = .003) and stricturing disease (odds ratio, 6.6; P = .0001) was more frequent and the risk for surgery was higher (hazard ratio, 5.8; P < .0001) and surgery occurred earlier (hazard ratio, 2.24) in those children with 3020insC mutation compared with those without 3020insC. CONCLUSIONS: In children with pediatric-onset CD, early development of stricturing behavior leading to surgical resection is influenced by ileal location and 3020insC variant of the CARD15 mutation. Genetic testing may identify children with CD who are at risk for early surgery. PMID- 15551254 TI - Clinical outcomes of double-balloon endoscopy for the diagnosis and treatment of small-intestinal diseases. AB - BACKGROUND & AIMS: A specialized system for a new method for enteroscopy, the double-balloon method, was developed. The aim of this study was to evaluate the usefulness of this endoscopic system for small-intestinal disorders. METHODS: The double-balloon endoscopy system was used to perform 178 enteroscopies (89 by the anterograde approach and 89 by the retrograde approach) in 123 patients. The system was assessed on the basis of the rates of success in jejunal and ileal insertion and the entire examination of the small intestine, diagnostic yields, ability to perform treatment, and complications. RESULTS: Insertion of the endoscope beyond the ligament of Treitz or ileocecal valve was possible in all 178 procedures. It was possible to observe approximately one half to two thirds of the entire small intestine by each approach, and observation of the entire small intestine was possible in 24 (86%) of 28 trials. The source of bleeding was identified in 50 (76%) of 66 patients with GI bleeding, scrutiny of strictures was possible in 23 patients, and a tumor was examined endoscopically in 17 patients. Two complications (1.1%) occurred. Endoscopic therapies in the small intestine including hemostasis (12 cases), polypectomy (1 case), endoscopic mucosal resection (1 case), balloon dilation (6 cases), and stent placement (2 cases) were performed successfully. CONCLUSIONS: Double-balloon endoscopy permits the exploration of the small intestine with a high success rate of total enteroscopy. The procedure is safe and useful, and it provides high diagnostic yields and therapeutic capabilities. PMID- 15551255 TI - Screening for defective DNA mismatch repair in stage II and III colorectal cancer patients. AB - BACKGROUND & AIMS: Colorectal cancers associated with the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome usually present in younger patients, show loss of mismatch repair (MMR) gene expression, and exhibit microsatellite instability (MSI). About 12% of sporadic colorectal cancers also show MMR loss and MSI. The aims of this study were to evaluate MMR loss and MSI in relation to patient age, sex, tumor stage, and site in the large bowel. METHODS: Tissue microarrays were created from 1020 stage II and III colorectal cancer cases and immunohistochemical staining performed to detect expression of the 2 major MMR proteins, hMLH1 and hMSH2. MSI was determined using the BAT-26 mononucleotide repeat. RESULTS: Ten percent of tumors showed loss of hMLH1 expression and 1.2% showed loss of hMSH2 expression. hMLH1 loss was more frequent in women (P < .001), older patients (P = .004), earlier stage tumors (P = .0001), and proximal colon tumors ( P < .0001). In contrast, tumors showing hMSH2 loss were more frequent in younger (P < .001), male (P = .05) patients and were distributed evenly between the proximal colon and distal colon/rectum. Eleven percent of tumors were MSI+ and these showed similar age, sex, stage, and site characteristics as tumors with hMLH1 loss. Discordance between MMR loss and MSI+ was found in 24 of 983 (2.4%) tumors. Of the 231 patients aged <60 years at diagnosis, 12 (5.2%) showed loss of hMLH1 and 8 (3.5%) showed loss of hMSH2. CONCLUSIONS: Routine immunohistochemical screening for MMR loss in younger colorectal cancer patients may provide a useful, first-step screening tool for the population-based detection of HNPCC. PMID- 15551256 TI - Pancreatic mucinous cystic neoplasm defined by ovarian stroma: demographics, clinical features, and prevalence of cancer. AB - BACKGROUND & AIMS: Pancreatic mucin-producing cystic neoplasms are classified into 2 distinct entities: mucinous cystic neoplasm (MCN) and intraductal papillary mucinous neoplasm (IPMN). In previous studies, MCN often has been defined loosely and has not always been distinguished clearly from IPMN. Our aims were to determine the demographics, clinical features, and prevalence of invasive cancer in MCN defined by the presence of characteristic ovarian stroma. METHODS: By using the presence of ovarian stroma as a requisite criterion for diagnosis of MCN, a single pathologist, unaware of clinical information, identified 56 MCNs from 243 mucin-producing neoplasms resected at Mayo Clinic between 1986 and 2003. Medical records of the MCN patients were reviewed to obtain clinical and demographic data. RESULTS: Patients with MCN were almost exclusively (98%) women; we identified 1 man with a neoplasm containing ovarian stroma. The mean (+/-SD) age at resection was 48 +/- 15 years (84% < 60 y). Abdominal pain was the most common presenting symptom; 16% were asymptomatic. Most MCN (93%) were in the pancreatic body/tail region. Their median size was 5 cm (61% > or =5 cm). Histologically, 50 (89%) were adenomas, 2 (4%) had carcinoma-in-situ, and 4 (7%) had invasive cancer. None of the 22 MCNs <5 cm in size had invasive cancer. No patient with noninvasive disease had a recurrence after resection. CONCLUSIONS: MCN defined by ovarian stroma has a distinct demographic and clinical profile and a low prevalence of invasive cancer. These observations suggest that ovarian stroma should be used as the defining criterion for diagnosing MCN. PMID- 15551257 TI - Population-based study of the epidemiology of and the risk factors for pyogenic liver abscess. AB - BACKGROUND & AIMS: Pyogenic liver abscess (PLA) is relatively uncommon in North America but is associated with significant morbidity and mortality. Our objective was to characterize the incidence of, risk factors for, and outcomes of PLA in a nonselected population. METHODS: Population-based surveillance was conducted in the Calgary Health Region (CHR) between April 1, 1999 and March 31, 2003. All adult CHR residents with PLA were identified, and charts were reviewed. RESULTS: Seventy-one CHR residents developed a PLA for an annual incidence of 2.3 per 100,000 population. There was an increasing incidence of PLA with advancing age. Men were at much higher risk of acquiring a PLA as compared to women (3.3 vs 1.3 per 100,000; relative risk [RR], 2.6; 95% confidence interval [CI], 1.5-4.6; P < .001), and this was observed across all age groups. A number of comorbid conditions were associated with significantly higher risk for developing a PLA including liver transplantation patients (RR, 444.8; 95% CI, 89.5-1356.0; P < .0001), diabetics (RR, 11.1; 95% CI, 6.3-19; P < .0001), and patients with a history of malignancy (RR, 13.3; 95% CI, 6.9-24.4; P < .0001). No other solid organ transplantation patient was at increased risk. All patients required admission to hospital (median length of stay, 16 days), and 7 (10%) patients died in hospital, corresponding to a mortality rate of 0.22 per 100,000 population. CONCLUSIONS: This study provides important data on the burden of PLA and identifies risk groups that might potentially benefit from preventive efforts. PMID- 15551258 TI - Endothelin-receptor antagonist treatment of portopulmonary hypertension. AB - BACKGROUND & AIMS: Portal hypertension leading to variceal bleeding and ascites is a severe complication of liver cirrhosis and is associated with a high mortality rate. Endogenous vasoconstrictors including endothelin might play a role in control of intrahepatic vascular tone. Few patients develop pulmonary hypertension in the setting of portal hypertension, known as portopulmonary hypertension. METHODS/RESULTS: In this case report, we describe a patient with portopulmonary hypertension in the setting of cryptogenic liver cirrhosis. The patient received the dual antagonist of receptors A and B of endothelin-1 bosentan. After 16 and 31 weeks the pulmonary arterial pressure dropped significantly from the baseline value of 88 mm Hg to 63 and 58 mm Hg, respectively. Hepatic venous portal gradient was measured before and after bosentan treatment. Hepatic venous portal gradient significantly decreased from 26 mm Hg (baseline) to 7 and 17 mm Hg at 16 and 31 weeks after treatment with bosentan, respectively. CONCLUSIONS: The present report demonstrates the efficacy of bosentan in the treatment of portopulmonary and portal hypertension in the clinical setting. There are ongoing prospective studies to confirm these data. PMID- 15551259 TI - A 2-bp deletion in the GJA1 gene is associated with oculo-dento-digital dysplasia with palmoplantar keratoderma. AB - Oculo-dento-digital dysplasia (ODDD, OMIM no. 164210) is a pleiotropic disorder characterized mainly by ocular anomalies, varying degrees of finger and toe syndactyly, and enamel defects. It is caused by missense mutations in the gene coding for the gap junction protein connexin 43 or GJA1. Other types of mutations have so far not been reported. Here we describe a Dutch kindred with ODDD showing a new symptom, palmoplantar keratoderma, and associated with a novel 2-bp deletion mutation of GJA1. The dinucleotide deletion 780_781delTG is located in the cytoplasmic C-terminal loop and leads to a frameshift. This is predicted to lead to the production of a slightly truncated protein with 46 incorrect amino acids in the C-terminal cytoplasmic loop (C260fsX307). This novel mutation may explain the presence of skin symptoms. PMID- 15551260 TI - Fatigue sensitivity of Y-TZP to microscale sharp-contact flaws. AB - The strength degrading effects of sharp-contact damage are examined for Y-TZP ceramic plates bonded to a plastic substrate. Contacts are made with Vickers and Berkovich diamond indenters at low loads (0.1 N to 100 N) in the ceramic lower surfaces prior to bonding to the substrates. The indentations remain in the subthreshold region, that is, without visible corner radial cracks, over the lower region (< 10 N) of the load range. A concentrated load is then applied sinusoidally to the ceramic upper surface, with the loading axis centered on the subsurface indentation flaw, thereby subjecting the flaw to cyclic tension. Relative to polished surfaces, the indentations diminish the single-cycle strength by an amount that increases with increasing indentation load. The critical number of cycles required to cause failure from the indentation flaws is then measured at specified maximum lower surface tensile stresses. At each indentation load, the strength of the ceramic plates diminishes with increased cycling. The scale of degradation is compared with that from previous studies on Y-TZP surfaces containing larger-scale surface damage: sandblast damage, as used in dental crown preparation; and blunt-contact trauma from a spherical indenter at 3000 N. These other damage modes are shown to be equivalent in their strength degrading capacity to diamond pyramid indentations in the microscale load range 0.1 N to 1 N, attesting to the highly deleterious nature of sharp particulate contacts. The mechanism of fatigue is considered in terms of microcrack evolution within the indentation damage zone. PMID- 15551261 TI - A panorama of lineage-specific transcription in hematopoiesis. AB - The hematopoietic system consists of more than ten differentiated cell types, all of which are derived from a single type of hematopoietic stem cell. The accessibility and interest of this system have made it a model for understanding normal and abnormal differentiation of mammalian cells. Newer techniques have generated a mass of data that requires integrative approaches for analysis and interpretation. The traditional view of the differentiation program holds that a small number of regulators are involved in each stage of cell specification. However, this may not be the case. Recent analyses have shown that almost all substantial subsets of genes, including the set of broadly expressed transcription factors, are expressed in patterns that are unique for each lineage. Further, much of this difference between lineages can be captured in two dimensional graphs. Understanding the biologic significance, mechanisms and constraints underlying these differences is a challenge for experimentalists and computational biologists alike. PMID- 15551262 TI - Genetics of tethered cord "syndrome": The FG syndrome. PMID- 15551263 TI - Strategies against food-borne diseases. PMID- 15551264 TI - PolyADP-ribose polymerase-1 (PARP-1) and the evolution of learning and memory. AB - PARP-1 is a multifunctional enzyme that can modulate gene expression. Cohen-Armon et al.(1) found that a homologue of PARP-1 is activated in the Aplysia nervous system as the animal responds to an aversive stimulus, which leads to sensitization, and during a more complex form of learning that involves feeding behavior. Significantly, inhibiting PARP-1 activation blocked the learning. Several key pathways in Aplysia neurons are activated both during learning and after injury, suggesting that mechanisms of learning evolved from primitive responses to injury. Since PARP-1 is evolutionarily conserved as a responder to various forms of stress, the finding that PARP-1 is activated during learning supports this idea. PMID- 15551265 TI - Genetic modules and networks for behavior: lessons from Drosophila. AB - Behaviors are quantitative traits determined through actions of multiple genes and subject to genome-environment interactions. Early studies concentrated on analyzing the effects of single genes on behaviors, often generating views of simplified linear genetic pathways. The genome era has generated a profound paradigm shift enabling us to identify all the genes that contribute to expression of a behavioral phenotype, to investigate how they are organized as functional ensembles and to begin to identify polymorphisms that contribute to phenotypic variation and are targets for natural selection. Recent studies show that the genetic architecture of behavior is determined by dynamic and plastic modular networks of pleiotropic genes and that the behavioral phenotype manifests itself as an emergent property of such networks. Such networks are exquisitely sensitive to genetic background and sex effects. This review describes how Drosophila can serve as a model for uncovering fundamental principles of the genetic architecture of behavior. PMID- 15551266 TI - Finally, a sense of closure? Animal models of human ventral body wall defects. AB - Malformations concerning the ventral body wall constitute one of the leading categories of human birth defects and are present in about one out of every 2000 live births. Although the occurrence of these defects is relatively common, few detailed experimental studies exist on the development and closure of the ventral body wall in mouse and human. This field is further complicated by the array of theories on the pathogenesis of body wall defects and the likelihood that there is no single cause for these abnormalities. In this review, we summarize what is known concerning the mechanisms of normal ventral body wall closure in humans and mice. We then outline the theories that have been proposed concerning human body wall closure abnormalities and examine the growing number of mouse mutations that impact normal ventral body wall closure. Finally, we speculate how studies in animal models such as mouse and Drosophila are beginning to provide a much-needed mechanistic framework with which to identify and characterize the genes and tissues required for this vital aspect of human embryogenesis. PMID- 15551267 TI - The evolution of the peculiarities of mammalian sex chromosomes: an epigenetic view. AB - In most discussions of the evolution of sex chromosomes, it is presumed that the morphological differences between the X and Y were initiated by genetic changes. An alternative possibility is that, in the early stages, a key role was played by epigenetic modifications of chromatin structure that did not depend directly on genetic changes. Such modifications could have resulted from spontaneous epimutations at a sex-determining locus or, in mammals, from selection in females for the epigenetic silencing of imprinted regions of the paternally derived sex chromosome. Other features of mammalian sex chromosomes that are easier to explain if the epigenetic dimension of chromosome evolution is considered include the relatively large number of X-linked genes associated with human brain development, and the overrepresentation of spermatogenesis genes on the X. Both may be evolutionary consequences of dosage compensation through X-inactivation. PMID- 15551268 TI - An extracellular driving force of cell-shape changes. AB - The cellular capacity to internalise objects, involving attachment, engulfment and uptake, exists in virtually all organisms. Many uptake reactions are associated with cell signalling. However, the mechanical forces that form endocytotic vesicles are not known. We propose a 'leverage-mediated' uptake mechanism involving lateral cross-linking processes on the cell surface that can generate the configurational energy to create an inverse curvature of the membrane. PMID- 15551269 TI - Insulin and its receptor: structure, function and evolution. AB - I present here a personal perspective on more than three decades of research into the structural biology of the insulin-receptor interaction. The solution of the three-dimensional structure of insulin in 1969 provided a detailed understanding of the insulin surfaces involved in self-assembly. In subsequent years, hundreds of insulin analogues were prepared by insulin chemists and molecular biologists, with the goal of relating the structure to the biological function of the molecule. The design of methods for direct receptor-binding studies in the 1970s, and the cloning of the receptor in the mid 1980s, provided the required tools for detailed structure-function studies. In the absence of a full three-dimensional structure of the insulin-receptor complex, I attempt to assemble the existing pieces of the puzzle generated by our and other laboratories, in order to generate a plausible mechanistic model of the insulin-receptor interaction that explains its kinetics and negative cooperativity. PMID- 15551270 TI - Hold the germ cells, I'm on duty. AB - Germ cell segregation and gamete production are developmental problems that all sexually reproducing species must solve in order to survive. Many people are familiar with the complex social structures of some insect species, where specialised castes of adult insects perform specific tasks, one of which is usually to guard the sexually reproductive queen. The parasitic wasp Copidosoma floridanum adds another level of complexity to the caste system: a fertilised egg produces both sterile, short-lived "soldier" larvae and "reproductive" larvae that complete metamorphosis to produce sexually reproductive adults. How two morphologically and functionally distinct larval castes are produced by genetically identical groups of cells developing under the same environmental conditions is a baffling problem. A recent paper suggests that differential germ cell segregation during embryogenesis may be an event both necessary and sufficient for caste determination.(1) PMID- 15551271 TI - Getting an embryo into shape. AB - Formation of a multicellular organism is a complex process involving differentiation and morphogenesis. During early vertebrate development, the radial symmetric organization of the egg is transferred into a bilateral symmetric organism with three distinct body axes: anteroposterior (AP), dorsoventral, and left-right. Due to cellular movements and proliferation, the body elongates along the AP axis. How are these processes coupled? Two recent publications now indicate that cell migration as well as orientated cell divisions contribute to axis elongation. The processes are coupled through the planar cell polarity pathway.1 At the same time, the AP axis is patterned independently of convergent extension. This process, however, is required for cell migration and represents a cue for polarized cell motility during gastrulation. Thus, it is AP polarity that instructs individual cells how to orientate with respect to the embryonic axis and provides positional information for the process of convergent extension.(2) PMID- 15551272 TI - Scent wars: the chemobiology of competitive signalling in mice. AB - Many mammals use scent marks to advertise territory ownership, but only recently have we started to understand the complexity of these scent signals and the types of information that they convey. Whilst attention has generally focused on volatile odorants as the main information molecules in scents, studies of the house mouse have now defined a role for a family of proteins termed major urinary proteins (MUPs) which are, of course, involatile. MUPs bind male signalling volatiles and control their release from scent marks. These proteins are also highly polymorphic and the pattern of polymorphic variants provides a stable ownership signal that communicates genome-derived information on the individual identity of the scent owner. Here we review the interaction between the chemical basis of mouse scents and the dynamics of their competitive scent marking behaviour, demonstrating how it is possible to provide reliable signals of the competitive ability and identity of individual males. PMID- 15551273 TI - Recombinational DNA repair: the ignored repair systems. AB - The recent finding of a role for the recA gene in DNA replication restart does not negate previous data showing the existence of recA-dependent recombinational DNA repair, which occurs when there are two DNA duplexes present, as in the case for recA-dependent excision repair, for postreplication repair (i.e., the repair of DNA daughter-strand gaps), and for the repair of DNA double-strand breaks. Recombinational DNA repair is critical for the survival of damaged cells. PMID- 15551274 TI - Shared components of protein complexes--versatile building blocks or biochemical artefacts? AB - Protein complexes perform many important functions in the cell. Large-scale studies of protein-protein interactions have not only revealed new complexes but have also placed many proteins into multiple complexes. Whilst the advocates of hypothesis-free research touted the discovery of these shared components as new links between diverse cellular processes, critical commentators denounced many of the findings as artefacts, thus questioning the usefulness of large-scale approaches. Here, we survey proteins known to be shared between complexes, as established in the literature, and compare them to shared components found in high-throughput screens. We discuss the various challenges to the identification and functional interpretation of bona fide shared components, namely contaminants, variant and megacomplexes, and transient interactions, and suggest that many of the novel shared components found in high-throughput screens are neither the results of contamination nor central components, but appear to be primarily regulatory links in cellular processes. PMID- 15551276 TI - Initial increase in the cryoglobulin level after rituximab therapy for type II cryoglobulinemia secondary to Waldenstrom macroglobulinemia does not indicate failure of response. AB - Type II (mixed) cryoglobulinemia is a systemic vasculitis that may affect many organs, including the skin, leading to ulcerations secondary to immune complex deposition. We report a patient who presented with a large necrotic ulcerative lesion on the right ankle secondary to type II cryoglobulinemic vasculitis due to Waldenstrom macroglobulinemia that was resistant to multiple modalities of therapy. Amputation to prevent further necrosis and infections was almost performed. Treatment with rituximab was initiated and the patient had a dramatic response. This case report highlights the effectiveness and prompt response to rituximab when used in patients with small vessel vasculitis and skin ulceration. We describe a rise in the cryoglobulin level that may occur initially after rituximab therapy. PMID- 15551277 TI - Efficacy of imatinib mesylate (STI571) in chronic neutrophilic leukemia with t(15;19): case report. AB - Chronic neutrophilic leukemia (CNL) is a rare hematologic disorder, for which there is no standard therapy. Recently, STI (imatinib mesylate) has been shown to be effective in treating patients with chronic myeloproliferative disorder (CMPD) displaying the translocation of the PDGFbetaR gene. Here, we present a case of a patient with CNL carrying t(15;19)(q13;p13.3) who achieved a cytogenetic remission following treatment with imatinib, 400 mg daily. After failure of alpha interferon and hydroxyurea therapy, a durable and complete clinical and cytogenic remission was induced by imatinib. To our knowledge, this is the first case with CNL who showed complete response with cytogenic remission after treatment of imatinib. The mechanism of response to this molecule is unknown in our case (other oncogenes than c-kit, tyrosine kinase, or PDGFR may be involved). The patient remains in complete remission with an excellent performance status after 7 months of therapy. We demonstrate here that imatinib can induce a clinical and cytogenetic response in a case of CNL associated with a novel translocation other than a 5q33 rearrangement. Further studies including the molecular cloning of the t(15;19)(q13;p13.3) will be important in understanding the pathophysiology of CNL with a heterogeneous clinical course and the exploitation of the basic mechanisms of imatinib treatment. PMID- 15551278 TI - Type I Gaucher disease with severe skeletal destruction, extraosseous extension, and monoclonal gammopathy. AB - Extraosseous extensions of Gaucher-cell deposits simulate malignant diseases. We describe a 65-year-old male with type-I Gaucher disease, confirmed by low leukocyte glucocerebrosidase activity, high plasma chitotriosidase, and N370/L444P gene-mutations, who had severe skeletal involvement, IgG-kappa monoclonal gammopathy, and a soft-tissue mass within the left iliac muscle. Bone marrow biopsy showed heavy infiltration by Gaucher cells, and histopathology of the excised extraosseous mass revealed infiltration by Gaucher cells. Thus, malignant diseases were excluded and the diagnosis of an extraosseous Gaucher cell extension was well documented. Our case is reported because it is very interesting and unique in the literature. PMID- 15551279 TI - Pericardial effusion: a rare presentation of adult T-cell leukemia/lymphoma. AB - Adult T-cell leukemia/lymphoma (ATLL) has a wide variety of clinical presentations. The most common ones include leukemia, hypercalcemia, lymphadenopathy, hepatosplenomegaly, and skin lesions. We report a case of ATLL in a 73-year-old woman who presented initially with chest discomfort and shortness of breath. The patient had no peripheral lymphadenopathy, circulating lymphoma cells, hepatosplenomegaly, or skin lesions. CT scan showed small mediastinal lymph nodes and pericardial effusion. Diagnosis was established by cytomorphologic evaluation and flow cytometric analysis of the pericardial fluid. Cardiac involvement is a rare event in ATLL and, when present, usually is a late finding in the setting of disseminated disease. This case was unusual because the patient lacked all common clinical features of ATLL. We present this case so as to increase awareness that ATLL could initially present with pericardial effusion. The pathophysiologic mechanisms of cardiac involvement are also discussed. PMID- 15551280 TI - Dissociation between the level of von Willebrand factor-cleaving protease activity and disease in a patient with congenital thrombotic thrombocytopenic purpura. AB - Decreased von Willebrand factor (VWF)-cleaving protease activity (<5%) has been implicated in patients with congenital thrombotic thrombocytopenic purpura hemolytic uremic syndrome (Upshaw-Schulman syndrome) and associated with mutations within the ADAMTS13 gene. In this report, we describe longitudinal studies in a patient with congenital TTP who ultimately developed end-stage renal failure and required plasma therapy from infancy. The patient was deficient in plasma high molecular weight (HMW)-VWF multimers during acute disease but had increased amounts of the HMW-VWF multimers during periods of remission. DNA analysis of this patient detected homozygosity for the R692C mutation on exon 17 of the ADAMTS13 gene, previously linked to congenital TTP. The level of VWF cleaving protease activity in the patient was remarkably low (<5%) throughout her disease, even after she entered complete remission. However, despite no improvement in the level of VWF-cleaving protease activity, this patient had complete resolution of disease following splenectomy and commencing hemodialysis, without need for ongoing plasma therapy. The patient has remained in remission for over 4 years. These observations suggest that there are other factors in conjunction with severe deficiency of VWF protease activity that participate in the platelet-mediated thrombotic complications and other disease manifestations of congenital TTP. In addition, it is possible that splenectomy could be an effective treatment option for some patients with severe, congenital TTP. PMID- 15551281 TI - Potential role of the ventilation and perfusion (V/Q) lung scan in the diagnosis of acute chest syndrome in adults with sickle cell disease. AB - The current criteria for diagnosing ACS are chest pain and presence of a new infiltrate on the chest radiograph (CXR). This study was designed to evaluate the role of ventilation and perfusion (V/Q) scan to assist in the early diagnosis of ACS. An abnormal V/Q scan was associated with a diagnosis of ACS that reached a statistical significance (P < 0.038). The sensitivity and specificity were found to be 60% and 100%, respectively. We conclude that V/Q scan may play a role in the early diagnosis of ACS. PMID- 15551282 TI - Stimulation of Kaposi's sarcoma-associated herpesvirus viremia during hematopoietic stem cell mobilization with filgrastim. AB - The effects of hematopoietic stem cell (HSC) mobilization on Kaposi's sarcoma associated herpesvirus (KSHV) were evaluated in three KSHV and human immunodeficiency virus type 1 co-infected subjects. KSHV DNA was not detected in purified CD34+ cell preparations from the period of filgrastim treatment. However, two of 3 subjects had transiently increased cell-free plasma KSHV DNA during filgrastim treatment. Peak plasma KSHV DNA (2,600 and 4,300 copies/mL) occurred on day 4 and declined to below the limit of detection by day 7. These findings suggest that, although CD34+ cell preparations do not have evidence of KSHV infection, HSC mobilization may stimulate KSHV replication in other cellular compartments that contribute to KSHV viremia. PMID- 15551283 TI - Cytarabine-induced neurotoxicity responding to methyl prednisolone. PMID- 15551284 TI - Plasma kinetics of a cholesterol-rich microemulsion in subjects with heterozygous beta-thalassemia. AB - Patients with beta-thalassemia trait have been reported to present lower plasma concentrations of low-density lipoprotein (LDL) and lower frequencies of acute myocardial infarction than normal subjects. In this study, the metabolism of LDL was tested in 12 patients with heterozygous beta-thalassemia trait (HBT) and 13 healthy subjects without the disease by determining the plasma kinetics of an artificially made cholesterol-rich microemulsion (LDE) that mimics the LDL metabolism and binds to LDL receptors. The emulsion was labeled with 14C cholesterol ester and injected intravenously into the subjects. Blood samples were drawn at regular intervals over 24 hr to determine the plasma decay curve of the emulsion radioactive label and to estimate its plasma fractional clearance rate (FCR, in hr(-1)). FCR of the 14C-cholesterol ester was greater in HBT compared to controls (0.0631 +/- 0.0178 hr(-1) and 0.0501 +/- 0.0094 hr(-1), respectively; mean +/- SD, P = 0.022). No differences were found regarding LDL cholesterol plasma concentration between the two groups, but apolipoprotein B concentration was lower in HBT than in control subjects (80 +/- 44 and 96 +/- 14, respectively; mean +/- SD, P = 0.026). Our results show that LDE FCR is increased in HBT, indicating that LDL clearance is increased in patients with beta thalassemia trait possibly due to the increased proliferation in the bone marrow of erythroid precursors. PMID- 15551285 TI - Methylenetetrahydrofolate reductase gene (MTHFR) polymorphisms and reduced risk of malignant lymphoma. AB - Folate and methionine are important nutrients in the "one-carbon" metabolism that is closely associated with DNA synthesis and DNA methylation. Genetic variation in these pathways may change susceptibility to cancer development. We have previously reported associations between lymphoma risk and germline polymorphisms in genes of methylenetetrahydrofolate reductase (MTHFR C677T and A1298C) and methionine synthase (MTR A2756G), finding the genotype other than the MTHFR 677CC/1298AA to confer a half-risk compared to the MTHFR 677CC/1298AA and a 3 fold higher risk with the MTR GG genotype than the AA/AG genotypes. To confirm the association and explore the histological difference, we extended the previous case series. A case-control study was conducted in Japan with a total of 372 lymphoma cases and 500 noncancer controls examined for genotypes. The relative risks were estimated by unconditional logistic regression analysis. In overall analyses, the age-sex adjusted odds ratio (OR) for the subjects harboring MTHFR 677T or 1298C alleles relative to 677CC/1298AA genotype was 0.58 (95% confidence interval: 0.41-0.83, P = 0.002). The MTR GG genotype showed an OR of 1.75 (0.87 3.52, P = 0.114). These findings were validated in separate analyses of the 273 new incident cases. Subgroup analyses according to histological subtype [diffuse large B-cell lymphoma (DLB), follicular lymphoma (FL), low-grade lymphoma of mucosa associated lymphoid tissue (MALT), and others] illustrated similar associations with certain exceptions for FL and MALT. Our data showed an association between the MTHFR polymorphisms and malignant lymphoma risk for all histological subtypes, although the extent of contribution of these polymorphisms may differ somewhat with histological subtype. Lack of association with MTR polymorphism was also confirmed. PMID- 15551286 TI - Response of factor V inhibitor to rituximab in a patient who received liver transplantation for primary biliary cirrhosis. AB - A 43-year-old patient developed factor V inhibitor 6 months after liver transplantation for primary biliary cirrhosis in association with Sjogren's syndrome/systemic lupus erythematosus. She suffered from ecchymoses in the lower extremities. The factor V inhibitor was eradicated after 10 weekly doses of 375 500 mg/m2 rituximab. PMID- 15551287 TI - Hemophagocytosis exacerbated by G-CSF/GM-CSF treatment in a patient with myelodysplasia. AB - We describe a 75-year-old man with neutropenia in whom bone marrow aspirate and biopsy demonstrated hemophagocytosis associated with myelodysplasia (MDS). Therapy with granulocyte-colony stimulating factor (G-CSF) and granulocyte monocyte-colony stimulating factor (GM-CSF) caused splenomegaly and severe thrombocytopenia, which recurred upon rechallenge. We propose that myeloid growth factors may be detrimental in patients with MDS-associated hemophagocytosis. PMID- 15551288 TI - An unusual presentation of primary malignant lymphoma of the spleen. PMID- 15551289 TI - Prevalence of hepatitis C virus infection in IgM-type monoclonal gammopathy of uncertain significance and Waldenstrom macroglobulinemia. PMID- 15551290 TI - Cytokine profile of sickle cell disease in Oman. AB - The aim of our study was to assess the cytokine profile of sickle cell disease (SCD) patients in steady state and in vaso-occlusive crisis (VOC). VOC has a complex nature, involving interactions between sickle red blood cells (RBC), the endothelium, and leucocytes. Endothelial damage due to recurrent adhesion of sickle RBCs may disrupt endothelial function, leading to altered cytokine release. It is therefore pertinent to study the cytokine profile of SCD patients in steady state and in crisis prior to exploring its contribution to vaso occlusive manifestations, since it is believed that an altered balance of proinflammatory and anti-inflammatory cytokines plays an important role in painful crisis. Cytokines including IL-1beta, IL-2, IL-4, IL-6, IL-8, TNF-alpha, and IFN-gamma were measured by commercially available ELISA kits in SCD patients (n = 60); in steady state (n = 26) and in painful crisis (n = 34) and compared with nonanemic age- and sex-matched normal Omani controls (n = 20). SCD patients in crisis showed elevated levels of TNF-alpha (P < 0.092) and IL-6 (P < 0.024) when compared with steady state. It was also observed that SCD patients in steady state showed a significant elevation in IL-1beta (P < 0.04), IL-6 (P < 0.0001), and IFN-gamma (P < 0.02) as compared to normal subjects. It is thus evident that both type I and type II cytokines are significantly altered in SCD patients. In steady state, type II proinflammatory cytokines are elevated, whereas in crisis, an additional augmentation of type I cytokines occurs, with persistent elevation of type II cytokines, emphasizing the role of perturbed endothelium and activated monocytes in the pathophysiology of vaso-occlusion in sickle cell crisis. PMID- 15551291 TI - Case of relapsed CD20(+) mixed-cellularity Hodgkin disease treated with sequential rituximab and radiotherapy. PMID- 15551292 TI - Resolution of acquired factor X deficiency with amyloidosis secondary to plasma cell dyscrasia. PMID- 15551293 TI - Thrombosis associated with L-asparaginase therapy and low fibrinogen levels in adult acute lymphoblastic leukemia. AB - L-asparaginase is a chemotherapeutic agent commonly used in the treatment of both adult and pediatric acute lymphoblastic leukemia (ALL). A major complication is thrombosis, resulting from reduced synthesis of proteins such as antithrombin III. Hypofibrinogenemia, also a side effect, may be a marker of thrombosis and decreased protein synthesis. A retrospective chart review of identically treated patients revealed 9 thrombotic events among 93 patients (10%), 6 (7%) occurring during treatment cycles including L-asparaginase. Twelve (13%) patients had fibrinogen levels <50 mg/dL. Of these, 3 (25%) suffered a thrombotic event. This results in a specificity of 90% and a relative risk of 10 (P = 0.014). Therefore, a fibrinogen <50 mg/dL may serve as a marker for a hypercoagulable state in ALL patients receiving L-asparaginase. PMID- 15551294 TI - Prognostic variables in newly diagnosed childhood immune thrombocytopenia. AB - Immune thrombocytopenia (ITP) has a favorable prognosis in children. Only a small number of children go on to develop chronic ITP. However, at the time of diagnosis, it is not possible to predict the course of the disease. In order to determine prognostic factors that could predict the disease course at diagnosis, we retrospectively evaluated various clinical variables in 103 pediatric patients with newly diagnosed ITP at our institution from 1995 to 2001. Sixty-eight (66%) patients had a mean platelet volume (MPV) of <8 fL on admission. Of 72 patients who had a follow-up period of at least 6 months, 54 (75%) achieved a durable remission within 6 months and 18 (25%) developed chronic ITP. In univariate analysis, a low admission MPV (<8), history of viral prodrome, and a low admission platelet count (<10 x 10(9)/L) predicted for a favorable outcome. Age and sex did not correlate with remission. In multivariate analysis, a low admission MPV and a history of a viral prodrome were the only independent factors correlated with a durable CR. The adjusted odds ratio for achieving a durable remission was 8.9 (95% CI: 1.54-51.8) for history of a viral prodrome and 14 (95% CI: 2.52-83.3) for low admission MPV value. In conclusion, our study showed that a majority of the children with newly diagnosed ITP presented with a low MPV value. A history of viral illness and a low admission MPV were found to be independent prognostic variables that predicted for the achievement of a durable CR in childhood ITP. PMID- 15551295 TI - Postprandial glucose monitoring in type 1 diabetes mellitus: use of a continuous subcutaneous monitoring device. AB - BACKGROUND: Pre-prandial glucose gives insufficient information on glycemic excursions throughout the day. We aimed to test a continuous subcutaneous glucose monitoring device (GlucoDay) to describe postprandial glucose changes. METHODS: In 23 T1DM patients, 24-h GlucoDay registrations were started about 14 h before receiving a standard breakfast B and 3 h later lunch L. RESULTS: The 3-min glucose values were computed into parameters describing the postprandial changes after B and L. Two-hour glucose was higher after B (243 +/- 69 vs 180 +/- 79 mg/dL after L, p < 0.0001). Maximum glycemia (313 +/- 105 mg/dL after B and 304 +/- 119 after L, p < 0.0001) was higher and was reached after 78 and 57 min respectively. Three-hour AUC was higher but 30-min AUC was lower after B (5725 +/ 2414 vs 7488 +/- 2208 min mg/dL after L, p = 0.004). Glucose spikes (maximum peak minus fasting plasma glucose) were similar after B and L but the difference between maximum and minimum values was smaller after B (165 +/- 110 vs 219 +/- 115 mg/dL after L, p = 0.020). Duration of hyperglycemic periods >200, 140 or 126 mg/dL were not different after B or L, but time spent at glucose <100 mg/dL was longer after L (p < 0.0001). CONCLUSIONS: These results illustrate the use of subcutaneous glucose registration to characterize postprandial glycemia patterns in T1DM. Application of such methods to evaluate this and other clinical situations in DM can lead to therapeutic and dietary adjustments and ultimately improve glycemic control. PMID- 15551296 TI - The relationship of postprandial glucose to HbA1c. AB - The gold standard for the assessment of the overall glycemic control is the determination of HbA1c. There are, however, insufficient data to determine reliably the relative contribution of fasting and postprandial plasma glucose to HbA1c. Increasing evidence suggests that excessive excursions of postprandial glucose might be important for the development of micro- and macroangiopathic complications. With respect to the treatment options, one important question to be answered is whether premeal, postmeal or fasting plasma glucose, alone or in combination, will be necessary in adjusting the therapy to achieve optimal HbA1c levels while minimizing hypoglycemia. HbA1c is difficult to predict from fasting plasma glucose. There are indications that there is a shift in the relative contribution from postprandial glucose at good to fair HbA1c levels (<7.3% to <9.2%) to fasting plasma glucose at high HbA1c (>9.3%). There is also a better correlation of afternoon and evening plasma glucose with HbA1c than with prebreakfast and prelunch plasma glucose values. Since the definition on how to define postprandial glucose is still a matter of debate and since postprandial glucose depends on the premeal blood glucose level and, on the time of the meal, its size and composition and the therapeutic strategy, the data so far available are inconclusive and the best correlation of HbA1c is with the area under the glucose profiles. Continuous glucose monitoring under daily life conditions will be the key to definitely unravel the relationship among HbA1c and fasting, premeal, postprandial and postabsorptive plasma glucose. PMID- 15551297 TI - Insulin therapy and hypoglycaemia: the size of the problem. AB - BACKGROUND AND METHODS: Hypoglycaemia is a fact of life for people with diabetes mellitus. Mild, asymptomatic episodes occur once or twice a week in insulin treated diabetic subjects. Asymptomatic hypoglycaemia, including nocturnal hypoglycaemia, occurs in about 25% of diabetic subjects treated with insulin therapy. Mild hypoglycaemia, if recurrent, induces unawareness of hypoglycaemia and impairs glucose counterregulation, which in turn predisposes to severe hypoglycaemia. Even brief hypoglycaemia can cause profound dysfunction of the brain. Prolonged, severe hypoglycaemia can cause permanent neurological sequels. In addition, it is possible that hypoglycaemia may accelerate the vascular complications of diabetes by increasing platelet aggregation and/or fibrinogen formation. Finally, hypoglycaemia may be fatal. Hypoglycaemia induced by insulin as treatment of type 1 diabetes mellitus (T1 DM) is not the consequence of diabetes, but invariably of the non-physiological replacement of insulin. RESULTS: A number of studies have demonstrated that by moving from non physiological to more physiological models of insulin therapy, most of the hypoglycaemia problems may be overcome, the percentage of glycated hemoglobin (A1c) decreased, and the quality of life improved. Interestingly, in T1 DM with hypoglycaemia unawareness, prevention of hypoglycaemia reverses not only unawareness but also improves glucose counterregulation, primarily the responses of adrenaline. CONCLUSIONS: In order to best prevent hypoglycaemia, insulin should preferably be given as continuous subcutaneous infusion via a minipump (the 'golden standard') or multiple daily insulin administrations with insulin analogues (basal insulin glargine, meal insulin rapid-acting insulin analogues) in T1 DM. PMID- 15551298 TI - Asymptomatic hypoglycaemia: identification and impact. AB - The Diabetes Control and Complications Trial (DCCT) demonstrated that intensive management in people with type 1 diabetes delays the onset and the progression of microvascular complications associated with the disease. However, it is also known that intensive regimens can increase the number of hypoglycaemias and that the perception of symptoms decreases in relation to the statutes of metabolic control 1. Impairment of hypoglycaemic awareness is reported by up to 50% of patients with type 1 diabetes and it is associated with an increase of severe episodes of hypoglycaemia 2. Self-monitoring of blood glucose (SMBG) has become a major tool in the management of diabetes. Current recommendations suggest frequent SMBG. The major inconvenience of SMBG is due to the limitations of the glycaemic profile obtained from intermittent finger-sticks. This is an incomplete picture of blood glucose excursions; moreover, the frequent SMBG is not readily accepted by patients suffering from diabetes because it is invasive and painful. PMID- 15551299 TI - Glucose control in diabetes. AB - Diabetes is a worldwide disease. Epidemiological data indicates that the degree and duration of hyperglycaemia is associated with the microvascular and macrovascular complications of disease. Glycated haemoglobin at baseline is a significant predictor of retinopathy, as well as proteinuria, amputation and survival. Intensive blood glucose control with either sulphonylureas, metformin or insulin substantially decreases the risk of microvascular complications. There are no thresholds within the diabetic range of blood glucose for risk of microvascular complications. To reduce risk, we should aim for normal blood glucose levels. Mortality correlates with several parameters of glucose control, but notably with the coefficient of variation of fasting glucose, a marker of glucose instability. Postprandial hyperglycaemia is also another independent risk factor for atherosclerosis. Glucose sensors have considerable potential for monitoring changes in blood glucose concentration. Protein glycation is widespread and glycation of haemoglobin (HbA1c) probably reflects level of glycation of other proteins. Levels of advanced glycation end products could be valuable in assessing the risk of progression to diabetic complications. The fact that a glycated protein, the glycated haemoglobin, is now widely used to estimate glucose control in diabetes emphasizes the importance of the impact of ambient blood glucose on protein levels, structure and function on the clinical outcome of diabetes. PMID- 15551300 TI - Nocturnal hypoglycaemia in type 1 diabetes--consequences and assessment. AB - Hypoglycaemia is inevitable when striving for low HbA1c values. Nocturnal hypoglycaemia often occurs without symptoms, but results in diminished next day well-being and hypoglycaemia unawareness. Frequency of nocturnal hypoglycaemia was first assessed in research ward settings, but suffered from insufficient glucose sampling frequency. This may have resulted in overestimation of the duration of hypoglycaemic episodes. The advent of the first continuous glucose sensor, the needle-type MedtronicMiniMed Continuous Glucose Measurement System, revolutionized the assessment of glucose values. However, on scrutiny, the first version of this sensor showed a drift into the hypoglycaemic area and delayed recovery from hypoglycaemia. Using the microdialysis-based GlucoDay system, our group reported a lower frequency of nocturnal hypoglycaemia in type 1 diabetes patients using an insulin pump, than that expected from the existing literature. Today, more than 80 years after the introduction of insulin for the treatment of type 1 diabetes, the associated frequency of nocturnal hypoglycaemia still awaits its definitive assessment. PMID- 15551302 TI - Survey of computer use for health topics by patients with head and neck cancer. AB - BACKGROUND: Computers are potentially powerful tools for patient education. E health, which refers to health services and information delivered through the Internet, is a growing phenomenon within the health-care field. We sought to describe computer use and interest in e-health resources among patients with head and neck cancer. METHODS: A questionnaire was administered to 207 patients with head and neck cancer attending oncology follow-up clinics at a single comprehensive cancer center. RESULTS: Forty-eight percent had never used a computer; 43% used one more than once a week. E-health information had been sought by 31%. Likelihood to access e-health information increased with education and income but decreased with age (p < or = .05). CONCLUSIONS: Many patients with head and neck cancer welcome information technology, but most prefer more traditional sources of information. Interventions to improve computer access and/or skills are largely undesired. Individuals seem to either embrace technology or not. In this respect, patients with head and neck cancer are similar to, rather than unique from, other patients with cancer. PMID- 15551303 TI - Mechanical properties of cast Ti-Hf alloys. AB - This study examined the mechanical properties of a series of Ti-Hf alloys. Titanium alloys with 10 to 40 mass % Hf were made with titanium and hafnium sponge in an argon-arc melting furnace. Specimens cast into magnesia-based investment molds were tested for yield strength, tensile strength, percentage elongation, and modulus of elasticity. Vickers microhardness was determined at 25 to 600 microm from the cast surface. X-ray diffractometry was also performed. Commercially pure Ti (CP Ti) and pure Ti prepared from titanium sponge were used as controls. The data (n = 5) were analyzed with a one-way ANOVA and the Student Newman-Keuls test (alpha = 0.05). The diffraction peaks of all the metals matched those for alpha Ti; no beta phase peaks were found. Alloys with Hf > or = 25% had significantly (p < 0.05) higher yield and tensile strength compared to the CP Ti and pure Ti. There were no significant differences (p > 0.05) in elongation among all the Ti-Hf alloys and CP Ti, whereas the elongation of alloys with Hf > or = 30% was significantly (p < 0.05) lower than that of the pure Ti. The cast Ti-Hf alloys tested can be considered viable alternatives to CP Ti because they were stronger than CP Ti and had similar elongation. PMID- 15551304 TI - AGG interspersion patterns in the CGG repeat of the FMR1 gene and linked DXS548/FRAXAC1 haplotypes in Brazilian populations. PMID- 15551305 TI - A disorder resembling pseudoachondroplasia but without COMP mutation. AB - Pseudoachondroplasia (PA) is an autosomal dominant skeletal dysplasia characterized by disproportionate short stature, generalized ligamentous laxity, irregular epi-metaphyseal ossification, and vertebral anomalies that regress with age. It usually manifests in the second year of life or later. The clinically and radiographically variable disorder is caused by mutations in the COMP gene. Parental gonadal mosaicism may lead to recurrence of the disorder in children of unaffected parents. Here, we describe sibs with bone changes similar to those seen in very severe PA born to clinically and radiographically unaffected parents. Sequencing of all 19 exons of the COMP gene failed to disclose a mutation. The sibs appear to be affected by a disorder resembling PA but resulting from a defect of an extracellular matrix protein other than COMP. It may be suspected in patients with unusually severe dwarfism, severe epi metaphyseal abnormalities, and persistent platyspondyly. PMID- 15551306 TI - Alterations of cancer-related genes in soft tissue sarcomas: hypermethylation of RASSF1A is frequently detected in leiomyosarcoma and associated with poor prognosis in sarcoma. AB - Aberrant methylation is a main mechanism of tumor suppressor gene inactivation in carcinogenesis. In this study, the methylation status of RASSF1A, p16, MLH1, MSH2 and ERalpha was investigated in 84 primary soft tissue sarcomas (STSs), including 22 liposarcomas, 18 malignant fibrous histiocytomas (MFHs), 18 leiomyosarcomas, 6 rhabdomyosarcomas, 6 neurogenic sarcomas and several other sarcoma entities. RASSF1A hypermethylation was detected in 17 of 84 (20%) STSs; however, methylation was more frequent in leiomyosarcomas (39%) compared to MFHs (6%; p < 0.015) and liposarcomas (18%). The p16 CpG island was methylated in 22 out of 82 (27%) cases. In 7 out of 81 (9%) STS samples, the promoter of MLH1 was methylated and in liposarcoma the methylation frequency was higher (14%). For MSH2, no hypermethylation was detected. Methylation of ERalpha was detected in 48 of 63 (76%) STSs, but also in 4 of 8 (50%) normal tissue samples. Furthermore, we analyzed mutational activation of K-ras and BRAF. In 4 out of 84 (5%) of STSs, a substitution at codon 599 of BRAF was found; however, no alteration of K-ras was detected. In an univariate Cox proportional-hazards regression model, we found that the risk of a tumor-related death for STS patients with methylated RASSF1A was significantly increased (RR = 2.9; p = 0.037). In summary, our data indicate that inactivation of RASSF1A is a common event in STS, especially in leiomyosarcoma. Thus, the methylation status of cancer-related genes was distinct in different STS and methylation of RASSF1A promoter can serve as prognostic marker in STSs. PMID- 15551307 TI - Biophysical measurement of brain tumor cohesion. AB - An advantage of using 3D multicellular spheres to study tumor biology is that they better approximate the interactions encountered by cells in vivo. Our previous studies have shown that the process of spheroid formation is governed by the same thermodynamic principles driving the formation of liquid droplets. This liquid-like behavior enables us to measure a key property influencing tumor behavior, namely, intercellular cohesion. We have developed a method, tissue surface tensiometry (TST), to measure the cohesivity, expressible as surface tension (sigma), of tissue aggregates under physiologic conditions. This study utilizes TST to measure the cohesivity of 3 widely used malignant astrocytoma cell lines of different in vitro invasive potentials. We compare invasiveness with aggregate cohesivity and with the expression of N-cadherin, a key mediator of cell-cell cohesion in neural tissues. We show that the cell lines exhibit liquid-like behavior since they form spheroids whose surface tension is both force- and volume-independent; that aggregates from each cell line have a distinct surface tension that correlates with their in vitro invasive capacity; that dexamethasone (Dex), a widely used therapeutic agent for the treatment of tumor-related cerebral edema, increases aggregate cohesivity and decreases invasiveness; that dexamethasone treatment decreases invasion in a dose-dependent manner but only when cells are in direct contact with one another; and that dex mediated decreased invasiveness correlates with increased aggregate cohesivity as measured by TST but not with N-cadherin expression or function. Our results demonstrate that for these cell lines, cohesivity is an excellent predictor of in vitro invasiveness. PMID- 15551308 TI - Bis(oxofluorenediyl)oxacyclophanes: synthesis, crystal structure and complexation with paraquat in the gas phase. AB - The first three representatives of the new family of oxacyclophanes incorporating two 2,7-dioxyfluorenone fragments, connected by [-CH(2)CH(2)O-](m) spacers (m=2 4), have been synthesized. The yield of the smallest oxacyclophane (m=2) is considerably higher with respect to the larger ones (m=3 and m=4), which are formed in comparable yields. Molecular modeling and NMR spectra analysis of the model compounds suggest that an essential difference in oxacyclophanes yields is caused by formation of quasi-cyclic intermediates, which are preorganized for macrocyclization owing to intramolecular pi-pi stacking interactions between the fluorenone units. The solid-state structures of these oxacyclophanes exhibit intra- and intermolecular pi-pi stacking interactions that dictate their rectangular shape in the fluorenone backbone and crystal packing of the molecules with the parallel or T-shape arrangement. The crystal packing in all cases is also sustained by weak C--HO hydrogen bonds. FAB mass spectral analysis of mixtures of the larger oxacyclophanes (m=3 and m=4) and a paraquat moiety revealed peaks corresponding to the loss of one and two PF(6) (-) counterions from the 1:1 complexes formed. However, no signals were observed for complexes of the paraquat moiety with the smaller oxacyclophane (m=2). Computer molecular modeling of complexes revealed a pseudorotaxane-like incorporation of the paraquat unit, sandwiched within a macrocyclic cavity between the almost parallel aligned fluorenone rings of the larger oxacyclophanes (m=3 and m=4). In contrast to this, only external complexes of the smallest oxacyclophane (m=2) with a paraquat unit have been found in the energy window of 10 kcal mol(-1). PMID- 15551309 TI - Attenuation of telomerase activity by hammerhead ribozyme targeting human telomerase RNA induces growth retardation and apoptosis in human breast tumor cells. AB - Ribozyme possesses specific endoribonuclease activity and catalyzes the hydrolysis of specific phosphodiester bonds, which results in the cleavage of target RNA sequences. Here, we evaluated the ability of hammerhead ribozymes targeting human telomerase RNA (hTR) to inhibit the catalytic activity of telomerase and the proliferation of cancer cells. Hammerhead ribozymes were designed against 7 NUX sequences located in open loops of the hTR secondary structure. We verified the ribozyme specificity by in vitro cleavage assay by using a synthetic RNA substrate. Subsequently, we introduced ribozyme expression vector into human breast tumor MCF-7 cells and assessed the biologic effects of ribozyme. Hammerhead ribozyme R1 targeting the template region of hTR efficiently cleaved hTR in vitro, and stable transfectants of this ribozyme induced the degradation of target hTR RNA and attenuated telomerase activity in MCF-7 cells. Moreover, the ribozyme R1 transfectant displayed a significant telomere shortening and a lower proliferation rate than parental cells. Clones with reduced proliferation capacity showed enlarged senescence-like shapes or highly differentiated dendritic morphologies of apoptosis. In conclusion, the inhibition of telomerase activity by hammerhead ribozyme targeting the template region of the hTR presents a promising strategy for inhibiting the growth of human breast cancer cells. PMID- 15551310 TI - Water-soluble molecular capsules: self-assembly and binding properties. AB - The self-assembly and characterization of water-soluble calix[4]arene-based molecular capsules (12) is reported. The assemblies are the result of ionic interactions between negatively charged calix[4]arenes 1 a and 1 b, functionalized at the upper rim with amino acid moieties, and a positively charged tetraamidiniumcalix[4]arene 2. The formation of the molecular capsules is studied by (1)H NMR spectroscopy, ESI mass spectrometry (ESI-MS), and isothermal titration calorimetry (ITC). A molecular docking protocol was used to identify potential guest molecules for the self-assembled capsule 1 a2. Experimental guest encapsulation studies indicate that capsule 1 a2 is an effective host for both charged (N-methylquinuclidinium cation) and neutral molecules (6-amino-2 methylquinoline) in water. PMID- 15551311 TI - Expeditious asymmetric synthesis of a stereoheptad corresponding to the C(19) C(27)-ansa chain of rifamycins: formal total synthesis of Rifamycin S. AB - In the presence of sulfur dioxide and an acid promoter, (-)-(1E,3Z)-2-methyl-1 ((1S)-1-phenylethoxy)penta-1,3-dien-3-yl isobutyrate reacts with (Z)-3 (trimethylsilyloxy)pent-2-ene giving a silyl sulfinate intermediate that undergoes, in the presence of palladium catalyst, a desilylation and retro-ene elimination of SO(2) with formation of (-)-(1Z,2S,3R,4S)-1-ethylidene-2,4 dimethyl-5-oxo-3-((1S)-1-phenylethoxy)-heptyl isobutyrate as major product. This ethyl ketone undergoes cross-aldol reaction with (2S)-2-methyl-3-[(tert butyldimethylsilyl)oxy]propanal giving an aldol that is reduced into a stereoheptad corresponding to the C(19)-C(27)-segment of Rifamycins with high diastereoselectivity and enantiomeric excess. PMID- 15551312 TI - Synthesis, structure, and nonlinear optical properties of cross-conjugated perphenylated iso-polydiacetylenes. AB - Monodisperse, cross-conjugated perphenylated iso-polydiacetylene (iso-PDA) oligomers, ranging from monomer 15 to pentadecamer 25, have been synthesized by using a palladium-catalyzed cross-coupling protocol. Structural characteristics elucidated by X-ray crystallographic analysis demonstrate a non-planar backbone conformation for the oligomers due to the steric interactions between alkylidene phenyl groups. The electronic absorption spectra of the oligomers show a slight red-shift of the maximum absorption wavelength as the chain length increases from dimer 17 b to pentadecamer 25, a trend that has saturated by the stage of nonamer 22. Fluorescence spectroscopy confirms that the pendent phenyl groups present on the oligomer framework enhance emission, and the relative emission intensity consistently increases as a function of chain length n. The molecular third-order nonlinearities, gamma, for this oligomer series have been measured via differential optical Kerr effect (DOKE) detection and show a superlinear increase as a function of the oligomer chain length n. Molecular modeling and spectroscopic studies suggest that iso-PDA oligomers (n>7) adopt a coiled, helical conformation in solution. PMID- 15551313 TI - Human papillomavirus type 18 variants: histopathology and E6/E7 polymorphisms in three countries. AB - In cervical cancer, human papillomavirus type 18 (HPV 18) and HPV 16 are predominantly related to adenocarcinomas (ADCs) and squamous cell carcinomas (SCCs), respectively. Here, we studied whether the geographically distributed HPV intratypic variants are also associated with histologically different tumors. A total of 44 HPV 18-positive and 91 HPV 16-positive cervical carcinomas from Indonesian, Surinamese and Dutch patients were histologically classified using hematoxilin and eosin, periodic acid Schiff plus and Alcian Blue staining. Samples were sequenced and intratypic variants were classified into the known phylogenetic branches. The Asian Amerindian HPV 18 variant was observed in 56% of ADCs compared to 15% of SCCs (p < 0.006). The African HPV 18 variant was exclusively found in SCCs. By sequencing the HPV 18 E6 and E7 open reading frames, we found predicted amino acid changes only in 8 samples. Two amino acid changes were consistent throughout the African branch. In HPV 16-positive tumors, we did not find a specific linkage between intratypic variants and histopathology. We conclude that HPV 18 intratypic variants are differentially associated with adenocarcinoma and squamous cell carcinoma of the cervix. The findings described here stress the biologic significance of intratypic HPV variants and might help explaining differences in the pathogenesis of cervical ADCs and SCCs. PMID- 15551314 TI - Six- and eightfold palladium-catalyzed cross-coupling reactions of hexa- and octabromoarenes. AB - Palladium-catalyzed sixfold coupling of hexabromobenzene (20) with a variety of alkenylboronates and alkenylstannanes provided hexaalkenylbenzenes 1 in up to 73 % and 16 to 41 % yields, respectively. In some cases pentaalkenylbenzenes 21 were isolated as the main products (up to 75 %). Some functionally substituted hexaalkenylbenzene derivatives containing oxygen or sulfur atoms in each of their six arms have also been prepared (16 to 24 % yield). The sixfold coupling of the less sterically encumbered 2,3,6,7,10,11-hexabromotriphenylene (24) gave the desired hexakis(3,3-dimethyl-1-butenyl)triphenylene (25) in 93 % yield. The first successful cross-coupling reaction of octabromonaphthalene (26) gave octakis-(3,3 dimethyl-1-butenyl)naphthalene (27) in 21 % yield. Crystal structure analyses disclose that, depending on the nature of the substituents, the six arms are positioned either all on the same side of the central benzene ring as in 1 a and 1 i, making them nicely cup-shaped molecules, or alternatingly above and below the central plane as in 1 h and 23. In 27, the four arms at C-1,4,6,7 are down, while the others are up, or vice versa. Upon catalytic hydrogenation, 1 a yielded 89 % of hexakis(tert-butylethyl)benzene (23). Some efficient accesses to alkynes with sterically demanding substituents are also described. Elimination of phosphoric acid from the enol phosphate derived from the corresponding methyl ketones gave 1-ethynyladamantane (3 b, 62 % yield), 1-ethynyl-1-methylcyclohexane (3 c, 85 %) and 3,3-dimethylpentyne (3 e, 65 %). 1 (Trimethylsilyl)ethynylcyclopropane (7) was used to prepare 1-ethynyl-1 methylcyclopropane (3 d) (two steps, 64 % overall yield). The functionally substituted alkynes 3 f-h were synthesized in multistep sequences starting from the propargyl chloride 11, which was prepared in high yields from the dimethylpropargyl alcohol 10 (94 %). The alkenylstannanes 19 were prepared by hydrostannation of the corresponding alkynes in moderate to high yields (42-97 %), and the alkenylboronates 2 and 4 by hydroboration with catecholborane (27-96 % yield) or pinacolborane (26-69 % yield). PMID- 15551315 TI - Car-Parrinello molecular dynamics study of the initial dinitrogen reduction step in Sellmann-type nitrogenase model complexes. AB - We have studied reduction reactions for nitrogen fixation at Sellmann-type model complexes with Car-Parrinello simulation techniques. These dinuclear complexes are especially designed to emulate the so-called open-side FeMoco model. The main result of this work shows that in order to obtain the reduced species several side reactions have to be suppressed. These involve partial dissociation of the chelate ligands and hydrogen atom transfer to the metal center. Working at low temperature turns out to be one necessary pre-requisite in carrying out successful events. The successful events cannot be described by simple reaction coordinates. Complicated processes are involved during the initiation of the reaction. Our theoretical study emphasizes two experimental strategies which are likely to inhibit the side reactions. Clamping of the two metal fragments by a chelating phosphane ligand should prevent dissociation of the complex. Furthermore, introduction of tert-butyl substituents could improve the solubility and should thus allow usage of a wider range of (mild) acids, reductants, and reaction conditions. PMID- 15551316 TI - Monomeric metal aqua complexes in the interlayer space of montmorillonites as strong Lewis acid catalysts for heterogeneous carbon-carbon bond-forming reactions. AB - Montmorillonite-enwrapped copper and scandium catalysts (Cu(2+)- and Sc(3+) monts) were easily prepared by treating Na(+)-mont with the aqueous solution of the copper nitrate and scandium triflate, respectively. The resulting Cu(2+)- and Sc(3+)-monts showed outstanding catalytic activities for a variety of carbon carbon bond-forming reactions, such as the Michael reaction, the Sakurai-Hosomi allylation, and the Diels-Alder reaction, under solvent-free or aqueous conditions. The remarkable activity of the mont catalysts is attributable to the negatively charged silicate layers that are capable of stabilizing metal cations. Furthermore, these catalysts were reusable without any appreciable loss in activity and selectivity. The Cu(2+)-mont-catalyzed Michael reaction proceeds via a ternary complex in which both the 1,3-dicarbonyl compound and the enone are coordinated to a Lewis acid Cu(2+) center. PMID- 15551317 TI - Synthesis of a new chiral source, (1R,2S)-1-Phenylphospholane-2-carboxylic acid, via a key intermediate alpha-phenylphospholanyllithium borane complex: configurational stability and X-ray crystal structure of an alpha monophosphinoalkyllithium borane complex. AB - A synthetic route to enantiomerically pure (1R,2S)-1-phenylphospholane-2 carboxylic acid (1), which is a phosphorus analogue of proline, has been established. A key step is the deprotonation-carboxylation of the 1 phenylphospholane borane complex 3 by using sBuLi/1,2-dipiperidinoethane (DPE). Configurational stability of the key intermediate, the amine-coordinated alpha phosphinoalkyllithium borane complex 4, was investigated by employing lithiodestannylation-carboxylation of both diastereomers of the 1-phenyl-2 trimethylstannylphospholane borane complex 7 in the presence of several kinds of amines, and as a result, 4 was found to be configurationally labile even at -100 degrees C. The key intermediate, the DPE-coordinated trans-1-phenyl-2 phospholanyllithium borane complex 9, was isolated, and the structure was identified by X-ray crystal structure analysis. This is the first X-ray crystal structure determined for an alpha-monophosphinoalkyllithium borane complex. Remarkably, the alkyllithium complex is monomeric and tricoordinate at the lithium center with a slightly pyramidalized environment, and the existence of a Li--C bond (2.170 A) has been confirmed. Moreover, (1)H-(7)Li HOESY and (6)Li NMR analyses suggested the structure of 9 in solution as well as the existence of an equilibrium between 9, its cis isomer, and the ion pair 8 at room temperature, which was extremely biased towards 9 at -100 degrees C. Finally, 1 was used as a chiral ligand in a palladium-catalyzed allylic substitution, and the desired product was obtained in high yield with good enantioselectivity. PMID- 15551318 TI - Ferrocenyl-modified DNA: synthesis, characterization and integration with semiconductor electrodes. AB - The ferrocenyl-nucleoside, 5-ethynylferrocenyl-2'-deoxycytidine (1) has been prepared by Pd-catalyzed cross-coupling between ethynylferrocene and 5-iodo-2' deoxycytidine and incorporated into oligonucleotides by using automated solid phase synthesis at both silica supports (CPG) and modified single-crystal silicon electrodes. Analysis of DNA oligonucleotides prepared and cleaved from conventional solid supports confirms that the ferrocenyl-nucleoside remains intact during synthesis and deprotection and that the resulting strands may be oxidised and reduced in a chemically reversible manner. Melting curve data show that the ferrocenyl-modified oligonucleotides form duplex structures with native complementary strands. The redox potential of fully solvated ferrocenyl 12-mers, 350 mV versus SCE, was shifted by +40 mV to a more positive potential upon treatment with the complement contrary to the anticipated negative shift based on a simple electrostatic basis. Automated solid-phase methods were also used to synthesise 12-mer ferrocenyl-containing oligonucleotides directly at chemically modified silicon <111> electrodes. Hybridisation to the surface-bound ferrocenyl DNA caused a shift in the reduction potential of +34 mV to more positive values, indicating that, even when a ferrocenyl nucleoside is contained in a film, the increased density of anions from the phosphate backbone of the complement is still dominated by other factors, for example, the hydrophobic environment of the ferrocene moiety in the duplex or changes in the ferrocene-phosphate distances. The reduction potential is shifted >100 mV after hybridisation when the aqueous electrolyte is replaced by THF/LiClO(4), a solvent of much lower dielectric constant; this is consistent with an explanation based on conformation-induced changes in ferrocene-phosphate distances. PMID- 15551319 TI - Hierarchical self-assembly of supramolecular spintronic modules into 1D- and 2D architectures with emergence of magnetic properties. AB - Hierarchical self-assembly of complex supramolecular architectures allows for the emergence of novel properties at each level of complexity. The reaction of the ligand components A and B with Fe(II) cations generates the [2x2] grid-type functional building modules 1 and 2, presenting spin-transition properties and preorganizing an array of coordination sites that sets the stage for a second assembly step. Indeed, binding of La(III) ions to 1 and of Ag(I) ions to 2 leads to a 1D columnar superstructure 3 and to a wall-like 2D layer 4, respectively, with concomitant modulation of the magnetic properties of 1 and 2. Thus, to each of the two levels of structural complexity generated by the two sequential self assembly steps corresponds the emergence of novel functional features. PMID- 15551320 TI - New chiral auxiliaries for dynamic kinetic resolution: from theory to experiment. AB - New efficient chiral auxiliaries for dynamic kinetic resolution (DKR) of bromides into amines are proposed, based on a theoretical rationalisation of known literature results. One example was synthesized and tested, affording diastereoselectivities up to 100%. Several results of DKR reactions are known, based on oxazolidinone or imidazolidinone units as chiral auxiliaries. Nevertheless, their behaviour was not fully understood until a recent paper that we published. We now used our proposed mechanism to rationalize the behaviour of other similar chiral auxiliaries and to propose small structure changes in imidazolidinone rings which could largely improve their performance. We could show that the good performance of these molecules as chiral auxiliaries for DKR reactions where bromine is the leaving group and a primary or secondary amine is the nucleophile is due, in a first step, to the formation of a hydrogen bond between the amine and the ring carbonyl oxygen and, in a second step, to the strong electrostatic interaction between the leaving bromide and the carbonyl oxygen in the C-3 substituent. Considering the behaviour of this substituent which rotates to minimize the electrostatic repulsion with the bromide when reaching the transition state, we proposed the introduction of a second substituent in the C-4 position of the imidazolidinone ring, which prevents such rotation, thus increasing the energy difference between the transition states of the two distereoisomers. With such an auxiliary we were able to increase the best de known in literature (88%), when benzylamine is used as nucleophile, to 99, or even 100%, when iodide replaces the bromide in the substrate. PMID- 15551321 TI - Molecular recognition and crystal energy landscapes: an X-ray and computational study of caffeine and other methylxanthines. AB - We introduce a new approach to crystal-packing analysis, based on the study of mutual recognition modes of entire molecules or of molecular moieties, rather than a search for selected atom-atom contacts, and on the study of crystal energy landscapes over many computer-generated polymorphs, rather than a quest for the one most stable crystal structure. The computational tools for this task are a polymorph generator and the PIXEL density sums method for the calculation of intermolecular energies. From this perspective, the molecular recognition, crystal packing, and solid-state phase behavior of caffeine and several methylxanthines (purine-2,6-diones) have been analyzed. Many possible crystal structures for anhydrous caffeine have been generated by computer simulation, and the most stable among them is a thermodynamic, ordered equivalent of the disordered phase, revealed by powder X-ray crystallography. Molecular recognition energies between two caffeine molecules or between caffeine and water have been calculated, and the results reveal the largely predominant mode to be the stacking of parallel caffeine molecules, an intermediately favorable caffeine water interaction, and many other equivalent energy minima for lateral interactions of much less stabilization power. This last indetermination helps to explain why caffeine does not crystallize easily into an ordered anhydrous structure. In contrast, the mono- and dimethylxanthines (theophylline, theobromine, and the 1,7-isomer, for which we present a single-crystal X-ray study and a lattice energy landscape) do crystallize in anhydrous form thanks to the formation of lateral hydrogen bonds. PMID- 15551322 TI - The versatile, efficient, and stereoselective self-assembly of transition-metal helicates by using hydrogen-bonds. AB - A diverse range of dinuclear double-stranded helicates in which the ligand strand is built up by using hydrogen-bonding has been synthesized. The helicates, formulated as [Co(2)(L)(2)(L-H)(2)X(2)], readily self-assemble from a mixture of a suitable pyridine-alcohol compound (L; for example, 6-methylpyridine-2 methanol, 1), and a CoX(2) salt in the presence of base. Nine such helicates have been characterized by X-ray crystallography. For helicates derived from the same pyridine-alcohol precursor, a remarkable regularity was found for both the molecular structure and the crystal packing arrangements, regardless of the nature of the ancillary ligand (X). A notable exception was observed in the solid state structure of [Co(2)(1)(2)(1-H)(2)(NCS)(2)] for which intermolecular nonbonded contacts between the sulfur atoms (SS=3.21 A) lead to the formation of 1D chains. Helicates derived from (R)-6-methylpyridine-2-methanol (2) are soluble in solvents such as CH(3)CN and CH(2)Cl(2), and their self-assembly could be monitored in solution by (1)H NMR, UV/Vis, and CD titrations. No intermediate complexes were observed to form in a significant concentration at any point throughout these titrations. The global thermodynamic stability constant of [Co(2)(2)(2)(2-H)(2)(NO(3))(2)] was calculated from spectrophotometric data to be logbeta=8.9(8). The stereoisomerism of these helicates was studied in some detail and the self-assembly process was found to be highly stereoselective. The chirality of the ligand precursors can control the absolute configuration of the metal centers and thus the overall helicity of the dinuclear assemblies. Furthermore, the enantiomers of rac-6-methylpyridine-2-methanol (3) undergo a self-recognition process to form exclusively homochiral helicates in which the four pyridine-alcohol units possess the same chirality. PMID- 15551324 TI - On the analyses of mixture vapor pressure data: the hydrogen peroxide/water system and its excess thermodynamic functions. AB - Reported here are some aspects of the analysis of mixture vapor pressure data using the model-free Redlich-Kister approach that have heretofore not been recognized. These are that the pure vapor pressure of one or more components and the average temperature of the complex apparatuses used in such studies can be obtained from the mixture vapor pressures. The findings reported here raise questions regarding current and past approaches for analyses of mixture vapor pressure data. As a test case for this analysis approach the H2O2-H2O mixture vapor pressure measurements reported by Scatchard, Kavanagh, and Tickner (G. Scatchard, G. M. Kavanagh, L. B. Ticknor, J. Am. Chem. Soc. 1952, 74, 3715-3720; G. M. Kavanagh, PhD. Thesis, Massachusetts Institute of Technology (USA), 1949) have been used; there is significant recent interest in this system. It was found that the original data is fit far better with a four-parameter Redlich-Kister excess energy expansion with inclusion of the pure hydrogen peroxide vapor pressure and the temperature as parameters. Comparisons of the present results with the previous analyses of this suite of data exhibit significant deviations. A precedent for consideration of iteration of temperature exists from the little known work of Uchida, Ogawa, and Yamaguchi (S. Uchida, S. Ogawa, M. Yamaguchi, Japan Sci. Eng. Sci. 1950, 1, 41-49) who observed significant variations of temperature from place to place within a carefully insulated apparatus of the type traditionally used in mixture vapor pressure measurements. For hydrogen peroxide, new critical constants and vapor pressure-temperature equations needed in the analysis approach described above have been derived. Also temperature functions for the four Redlich-Kister parameters were derived, that allowed calculations of the excess Gibbs energy, excess entropy, and excess enthalpy whose values at various temperatures indicate the complexity of H2O2-H2O mixtures not evident in the original analyses of this suite of experimental results. PMID- 15551323 TI - From a theoretical concept to biochemical reactions: strain-induced bond localization (SIBL) in oxidation of vitamin E. AB - The regioselectivity of the oxidation of alpha-tocopherol (the main component of vitamin E) to an ortho-quinone methide (oQM) has been explained in the literature mostly by the ill-defined term "Mills-Nixon effect". In this paper we describe the preparation of eleven alpha-tocopherol derivatives, different from each other by the sum of annulation angles, but keeping the electronic factors unchanged. These compounds underwent Ag(2)O oxidation, forming two isomeric oQMs that were trapped by vinylmethyl ether. It was found that the isomeric product ratio changes smoothly as a function of the annulation angles, not abruptly from one regioisomer to the other on going from five- to six- and seven-membered rings, as predicted by the Mills-Nixon effect. The relative amounts of the products were determined at four different temperatures, and assuming that the product ratio represents the relative rates ratio, the relative enthalpy of activations could be obtained. Theoretically (at B3LYP/6-31G* theoretical level) four different intermediates were considered. Each of these underwent angular angles deformations to model the two regioisomers. At each deformation angle the energy difference between the two intermediates models was correlated to the experimental data for each of the four intermediates. It was found that the angle deformed lithium (6-methyl-2-benzylium)phenolate correlated best (R>0.994) to the experimental data. This study confirms that the regioselectivity of the two isomeric oQMs in the oxidation of alpha-tocopherol and related compounds is simply a function of angular strain, best explained by the SIBL (strain-induced bond localization) model. In addition, this study provides evidence that the highest energy intermediate in the oxidation of vitamin E is a phenolate-benzyl cation. PMID- 15551325 TI - Altered expressions of HOX genes in human cutaneous malignant melanoma. AB - HOX genes act as master genes to control morphogenesis. In human, HOX genes form 4 clusters composing 9 to 11 HOX genes (39 genes in total) on different chromosomes. We hypothesized that aberrant expression of HOX genes was associated with development and subsequent progression of melanoma and that the 39 HOX gene expression pattern determined the sites where melanoma grew. The expression levels of 39 HOX genes in 15 human cutaneous melanoma specimens and 7 nevus pigmentosus specimens were quantified by a comprehensive analysis system based on the real-time RT-PCR method. We found that the expression levels of HOXA11, A13, B9, D12 and D13 in melanoma were higher than those in nevus pigmentosus and that the expression levels of HOXA11, B2 and C13 were significantly different between pT4 melanoma and pT1 to pT3 melanoma. It was most notable that the expression levels of HOXA1, A2, C4 and B13 in melanoma with distant metastasis were higher than those in melanoma without it. On the other hand, we found no relationship between HOX genes expression patterns and the growing sites of melanoma. These results indicated that the misexpressions of some specific HOX genes were implicated in melanoma genesis and metastasis but had no linkage with melanoma sites. PMID- 15551326 TI - Proximity of first intercourse to menarche and the risk of human papillomavirus infection: a longitudinal study. AB - Cross-sectional studies have suggested that compared with women who delay the start of their sexual career, those who first have intercourse soon after menarche are more susceptible to cervical human papillomavirus (HPV) infection and thus have a greater risk of cervical neoplasia. We describe, using longitudinal observations, how the risk of infection with HPV varies with the interval between menarche and first intercourse in 474 women aged 15-19 recruited within 12 months of first intercourse and before the acquisition of a second sexual partner. One hundred forty-five women became HPV-positive; the cumulative risk of HPV infection 3 years after first intercourse was 45.0% (95% CI = 37.9 51.2). In univariate analyses, the hazards ratio (HR) of HPV infection increased significantly with age at first intercourse (HR = 1.212 per year; 95% CI = 1.050 1.398), partner age (HR = 1.084 per year; 95% CI = 1.045-1.125) and when women reported a sexually experienced partner (HR = 2.794; 95% CI = 1.804-4.326); the interval between menarche and first intercourse was a significant predictor of infection, with an increase in the HR of 12.9% for every year of increase in this interval (95% CI = 2.1%-24.9%). In a multivariate analysis, compared with women who first had intercourse within 3 years of menarche, those who postponed first intercourse beyond this time had a greater risk of infection (HR = 1.581; 95% CI = 1.113-2.245) after controlling for age and sexual experience of partner. PMID- 15551327 TI - Craniofacial dyssynostosis: description of the first four Spanish cases and review. AB - Craniofacial dyssynostosis (CD) is characterized by premature fusion of the lambdoid and posterior part of the sagittal sutures, and short stature. Thus, the skull shape becomes dolichocephalic with protuberant forehead and either bulging or flat occiput. Facial changes are secondary to the skull defects, and some additional findings have also been described. We report on the first four known Spanish patients. They were unrelated and had Spanish ancestors. In the three previous reports about this syndrome, the authors hypothesized that the frequency of the gene causing CD must be rather high in the Spanish population, and relatively common in areas with Spanish ancestry. We have estimated the minimal birth prevalence of the syndrome in 0.51 per million livebirths. It has been previously suggested that the syndrome is inherited as an autosomal recessive trait, since there were two affected sisters among the nine published cases. Phenotypic variability is discussed in detail in this paper. We also underline several aspects for the anticipatory guidance of affected individuals, especially recommending a neurologic evaluation taking into account the radiologic findings in order to plan early interventions to avoid undesirable consequences of craniosynostosis. It is also recommended to perform additional studies (ophthalmologic, cardiologic, among others) to rule out the existence of associated anomalies, which are more frequent than previously considered. PMID- 15551328 TI - Familial lipomyelomeningocele: a further report. AB - Lipomyelomeningocele is a form of spina bifida occulta with a distinct pathogenesis that differentiates it from open neural tube defects. Familial forms are rare and the condition may be polygenic. We report on 2 affected siblings with similar lesions and raise the possibility of an autosomal recessive pattern of inheritance with implications for genetic counselling. PMID- 15551329 TI - Induced expression of human CCND1 alternative transcripts in mouse Cyl-1 knockout fibroblasts highlights functional differences. AB - Splicing of human cyclin D1 (CCND1) mRNA producing transcripts a and b is modulated by a common polymorphism (A --> G) located in a conserved splice donor region at nucleotide 870. CCND1 A/G(870) genotype is associated with tumour progression and clinical outcome in a variety of cancers. Although in vitro expression of cyclin D1 transcript a (CCND1(tra)) has been widely investigated, few studies have examined the expression of CCND1 transcript b (CCND1(trb)). We have studied the effects of inducible expression of human CCND1(trb) in comparison with human CCND1(tra) in a mouse fibroblast knock-out for cyclin D1 (MEF(Cyl-1-/-)). Inducible expression was in stable clones isolated from MEF(Cyl 1-/-) transfectants. Induction of CCND1(tra) produced a 36-kDa protein, which led to a significant increase in the proportion of cells in S-phase, as detected by BrdU incorporation after 32 hr, compared to non-induced cells (p = 0.012). Clones induced to express CCND1(tra) exhibited a significantly increased ability to grow in serum depleted (2% FCS) medium compared to non-induced clones (p = 0.0004). Induced expression of CCND1(trb) in MEF(Cyl-1-/-) transfectants produced a 31-kDa protein and resulted in no significant difference in DNA synthesis, neither did the cells acquire the ability to grow in serum-depleted conditions compared to non-induced cells. Induction of CCND1(trb) significantly enhanced the ability of MEF(Cyl-1-/-) transfectants to form colonies in soft agar, (average 30-fold increase) compared to non-induced clones or those induced to express CCND1(tra). Our data supports the emerging view that CCND1 alternate transcripts encode proteins with differing independent biological functions. We suggest that CCND1(tra) encodes a protein involved in regulating mitogen responsive, anchorage dependent G(1) progression, whereas CCND1(trb) modulates the ability of the cell to grow in an anchorage-independent manner. PMID- 15551330 TI - hOGG1 Ser326Cys polymorphism and G:C-to-T:A mutations: no evidence for a role in tobacco-related non small cell lung cancer. AB - Human 8-oxoguanine DNA glycosylase 1 (hOGG1) plays a major role in the repair of 8-hydroxyguanine, one of the major forms of DNA damage generated by reactive oxygen species in tobacco smoke. If left unrepaired by hOGG1, 8-hydroxyguanine can produce G:C-to-T:A transversions. Recent studies have suggested that the hOGG1 Ser326Cys polymorphism is associated with both a decrease in enzyme activity and an increased risk of lung cancer. To define the interaction between tobacco carcinogens, hOGG1-mediated DNA repair and DNA damage, we examined the role of the hOGG1 Ser326Cys polymorphism in mutation of the p53 gene in non small cell lung cancer (NSCLC). Tumor and nonneoplastic DNA were collected from 141 cigarette smokers with NSCLC. p53 mutations were detected by direct dideoxy sequencing and/or the GeneChip p53 assay in 74 of the 141 (52%) tumors. hOGG1 codon 326 polymorphisms were identified by polymerase chain reaction-restriction fragment length polymorphism analysis. The distribution of hOGG1 codon 326 genotypes was Ser/Ser, 90 of 141 (64%); Ser/Cys, 45 of 141 (32%); and Cys/Cys, 6 of 141 (4%). p53 mutations were significantly (p = 0.04) less common in NSCLC from patients with codon 326 Ser/Cys or Cys/Cys genotypes (21 of 51; 41%) than in NSCLC from Ser/Ser homozygotes (53 of 90; 59%). The decrease in p53 mutation frequency among carriers of the Cys allele was more evident in lung squamous cell cancer [7 of 17 (41%) for Cys/Cys and Ser/Cys vs. 27 of 38 (71%) for Ser/Ser; p = 0.04] than in nonbronchoalveolar adenocarcinoma [11 of 26 (42%) for Cys/Cys and Ser/Cys vs. 20 of 35 (57%) for Ser/Ser; p = 0.25]. The prevalence of G:C-to-T:A transversions was similar among hOGG1 codon 326 genotypes. In summary, the hOGG1 codon 326 Cys allele was associated with a decrease in p53 mutations and no effect on G:C-to-T:A transversions in NSCLC. This decrease in p53 mutations in vivo is not consistent with a decrease in the repair of 8-hydroxyguanine among carriers of the hOGG1 codon 326 Cys allele in vitro. PMID- 15551331 TI - Sydney Multicenter Study of Parkinson's disease: non-L-dopa-responsive problems dominate at 15 years. AB - One-third of the 149 people recruited 15 to 18 years ago in the Sydney Multicenter Study of Parkinson's disease have survived. The original study compared low-dose levodopa with low-dose bromocriptine. We now report the problems experienced by people who survive 15 years from diagnosis. The standardized mortality ratio is significantly elevated at 1.86 and is not significantly different between treatment arms. Falls occur in 81% of patients, and 23% sustained fractures. Cognitive decline is present in 84%, and 48% fulfill the criteria for dementia. Hallucinations and depression are experienced by 50%. Choking has occurred in 50%, symptomatic postural hypotension in 35%, and urinary incontinence in 41%. No patient is still employed, and 40% of patients live in aged care facilities. Although approximately 95% have experienced L-dopa-induced dyskinesia/dystonia and end of dose failure of medication, in the majority, these symptoms are not disabling. Dyskinesia and dystonia were delayed by early use of bromocriptine, but end-of-dose failure appeared at a similar time once L-dopa was added. The rate of disease progression is similar in both arms of the study. We conclude that the most disabling long-term problems of Parkinson's disease relate to the emergence of symptoms that are not improved by L-dopa. Neuroprotective interventions in Parkinson's disease should be judged by their ability to improve non-L-dopa-responsive aspects of the disease, rather than just by their capacity to delay the introduction of L-dopa or reduce its associated side effects. PMID- 15551332 TI - A novel mutation (E767K) in the second extracellular loop of the calcium sensing receptor in a family with autosomal dominant hypocalcemia. AB - Autosomal dominant hypocalcemia resulting from gain-of-function mutations of the calcium sensing receptor (CASR) is a rare familial disorder that can become evident at any age. We report a novel mutation (E767K) of the CASR in a family with autosomal dominant hypocalcemia. Ten members of the family had a history of hypocalcemia. The index case exhibited marked hypocalcemia and seizures in the newborn period, while her father who also has hypocalcemia, was largely asymptomatic except for a myocardial infarction-like event at 21 years of age, a new presentation of the disorder. The E767K mutation, which resides in the second extracellular loop adjacent to the fifth transmembrane domain, co-segregated with hypocalcemia in these two individuals. Both subjects are heterozygous for the mutation. The proband is also heterozygous for the previously reported CASR polymorphism of G990R in the intracellular domain, while her father is homozygous. The co-segregation of this naturally occurring mutation with autosomal dominant hypocalcemia supports the previously reported experimental model in which it was proposed that the three acidic residues (767, 758, and 759) in exo-loop 2 in CASR help maintain an inactive conformation of the receptor. PMID- 15551333 TI - Mapping nonverbal IQ in young boys with fragile X syndrome. AB - This study examines the developmental changes in nonverbal intellectual functioning evident in males with fragile X syndrome (FXS) measured by the Leiter International Performance Scales-Revised (Leiter-R). The Leiter-R provides both IQ scores and associated growth scores which permit the examination of both age based IQ scores and overall intellectual growth. Participants were 45 males with full mutation FXS and ranged in age from 4.0 to 13.8 years. Each child was assessed annually using the Leiter-R as part of a larger longitudinal battery for an average of 3.5 assessments per child and a range of 2-6 assessments, representing a total of 156 assessment occasions. Longitudinal analyzes of Leiter scores consisted primarily of hierarchical linear modeling, with the impact of chronological age, maternal education, fragile X mental retardation 1 protein (FMRP), autistic behaviors also being assessed. Findings revealed a significant linear decline in nonverbal IQ scores, with no effects of maternal education, autistic behaviors, or FMRP on mean level or rate of change in IQ scores over time. The decline slowed significantly around 8 years of age, but scores continued to decline into the 12th year of age. In contrast, a significant linear increase was observed in Leiter-R growth scores, which was negatively influenced by autistic behaviors. The rate of increase did not change over time, and neither mean level nor rate of increase was influenced by maternal education or FMRP levels. These findings suggest that declines in IQ are the result of steady, but suboptimal intellectual growth, rather than a true deterioration in overall intellectual functioning. PMID- 15551334 TI - Fetal presentation of PHACES syndrome. PMID- 15551335 TI - Familial hyper- and hypopigmentation with age-related pattern change. PMID- 15551336 TI - Mosaic trisomy 22: report of a patient with normal intelligence. PMID- 15551337 TI - Novel COL9A3 mutation in a family with multiple epiphyseal dysplasia. AB - Multiple epiphyseal dysplasia (MED) is a common skeletal dysplasia characterized by mild to moderate short stature, early-onset of osteoarthritis (OA) mainly in the hip and knee joints, and abnormally small and/or irregular epiphyses. MED is clinically and genetically heterogeneous. Six causative genes of MED have been reported, including type IX collagen genes (COL9A1, COL9A2, COL9A3). All the type IX collagen mutations previously reported cause exon skipping that loses the COL3 domain. Here we have identified a novel COL9A3 mutation co-segregating in a three generation family with MED. The mutation (IVS3 + 5G > A) was speculated to lose the COL3 domain by skipping of exon 3, which was confirmed by in vitro analysis. The patients were of normal height and had minimal complaints with phenotypes being more severe in male patients. The radiographic phenotypes of the patients were relatively milder than those of previously reported cases, and were indistinguishable to common, idiopathic OA. PMID- 15551338 TI - Novel case of dup(3q) syndrome due to a de novo interstitial duplication 3q24 q26.31 with minimal overlap to the dup(3q) critical region. AB - Dup(3q) syndrome is characterized by typical facial features, mental and growth retardation, often with congenital heart defects. The syndrome has attracted special attention because of the clinical overlap with Cornelia de Lange syndrome (CDLS). Patients with dup(3q) syndrome are trisomic for segments of the long arm of chromosome 3, most often within the region 3q21 to 3qter. Most cases have arisen as unbalanced translocations and do involve other chromosomes also. A dup(3q) minimal region has been defined at 3q26.3-q27. We report here a 15-month old boy with a de novo interstitial inverted duplication of 3q24-q26.31. Clinical evaluation revealed mild but typical features of dup(3q) syndrome. The duplication was characterized by conventional and molecular cytogenetics. The results allow further narrowing of the dup(3q) critical region at its distal end and suggest the existence of one or several major genes responsible for the dup(3q) syndrome in the proximal half of 3q26.31. Moreover, the results of fluorescence in situ hybridization (FISH) analysis with BAC probes suggest a disruption of the NLGN1 gene at the distal end of the duplication in 3q26.31 in the patient. The breakpoint within NLGN1 is unique for this patient, and the contribution of NLGN1 disruption to the phenotype of this patient remains unclear. Yet since NLGN1 is involved in synaptogenesis in the central nervous system, altered gene dosage is a good candidate for mental retardation as a recurrent feature of dup(3q) syndrome. PMID- 15551339 TI - Congenital stationary night blindness: report of an autosomal recessive family and linkage analysis. AB - Congenital stationary night blindness (CSNB) is a group of rare, non-progressive conditions of the retina characterized by abnormal rod function causing impaired night vision. Among them, the Schubert-Bornschein subgroup, itself divided into a complete and an incomplete form, is characterized by a specific electrophysiological pattern. Complete, Schubert-Bornschein CSNB is usually transmitted as a monogenic trait, and most familial cases result from mutations of the NYX gene located on the X chromosome. We report a very rare family with consanguineous, first-cousin parents, where a son and a daughter are affected with this condition, indicating autosomal recessive inheritance. As the family was too small for genome-wide linkage, we considered several candidate loci, including the sidekick SDK1 and SDK2 genes. The latter determine lamina-specific connectivity in the retina, a histological substrate of the ON pathway implicated in complete, Schubert-Bornschein CSNB. Although linkage was excluded in our family, observations like the present one may lead to the identification of a new molecular cause for this condition. PMID- 15551340 TI - Recurrent adjacent-2 segregation of a familial t(14;21)(q11.2;q11.2): phenotypic comparison of two brothers and a paternal aunt inheriting the der(14). AB - A 14-year-old boy was referred for a genetics evaluation after high-resolution chromosome analysis showed a small amount of extra material in the proximal long arm of chromosome 21. Five years prior, his karyotype analysis was interpreted as normal with a variant chromosome 21. The patient has short palpebral fissures, strabismus, flat antihelices of the ears, long thumbs with bilaterally absent interphalangeal creases, proximal bilateral 3/4 syndactyly, small testes, hypotonia, mental retardation, and speech problems. He has significant depression and behavioral problems including hyperactivity, aggression, and impulsivity. His 8-year-old brother has more severe behavioral disturbances and depression, but less significant mental retardation. A paternal aunt has mental retardation, is unusually docile, and appears similar to our patient. Chromosome analysis and fluorescence in situ hybridization (FISH) whole chromosome paint of chromosome 21 showed that the patient's father carries a "cryptic" balanced translocation, 46,XY, t(14;21)(q11.2;q11.2), as does the patient's paternal grandmother. Uniparental disomy studies using seven informative polymorphic nucleotide repeat markers from 14q and 21q confirmed biparental inheritance of the number 14 and 21 chromosomes for each brother, and indicate that they and the paternal aunt, all of whom inherited the der(14), are monosomic for proximal 21q and trisomic for proximal 14q. These karyotypes arose through an adjacent-2 segregation in the father on two occasions, and from the paternal grandmother on one occasion. This family is an example of recurrent malsegregation with translocations involving the acrocentrics. PMID- 15551341 TI - Continuous glucose monitoring in conditions other than diabetes. AB - BACKGROUND: The development of new systems for continuous glucose monitoring has recently increased the interest for their potential applications among physicians involved in diabetes care. One of the most common applications of such devices is the identification of hypoglycaemic events in insulin-treated diabetic patients (particularly during the night) and the evaluation of the full daily glucose excursions. METHODS: Among commercially available glucose sensors, the Glucoday system has been utilized for practical clinical application in the last two years. One of the most important features of this device is the accuracy in monitoring interstitial glucose values, specifically in the hypoglycaemic range. This feature is clinically relevant when applied in the clinical setting of patients with type 1 diabetes mellitus. The ability to monitor glucose continuously could be indeed a useful tool for the study of hypoglycaemic conditions other than diabetes. RESULTS: In patients with hyperinsulinaemic hypoglycaemia, recurrent episodes of asymptomatic hypoglycaemia are common, and in patients with glycogen storage diseases, avoidance of recurrent and prolonged hypoglycaemic episodes usually require frequent determinations by mean of home blood glucose monitoring. CONCLUSIONS: Experimental preliminary evidences suggest that this new technology could be applied in the clinical setting to help the physician to identify mainly nocturnal hypoglycaemic events, otherwise not revealed by traditional self blood-glucose monitoring, even in those patients who are not treated by conventional insulin therapy. PMID- 15551342 TI - Postprandial hyperglycaemia in type 2 diabetes: pathophysiological aspects, teleological notions and flags for clinical practice. AB - Type 2 diabetes subjects carry an excess risk for micro- and macrovascular disease and a higher cardiovascular morbidity and mortality rate. The beneficial impact of tight glycaemic control-evidenced by the integrated marker of fasting glucose and postprandial glucose values, the HbA1c-for the prevention of microvascular complications is definitely confirmed. Over the past few years, several studies have identified postprandial hyperglycaemia as a better predictor of cardiovascular or even of all-cause mortality, as well as an independent risk factor for atherosclerosis. The continuous glucose monitoring could offer a rationale means for the detection of postprandial hyperglycaemia and ultimately for its effective management. Advances in technology keep a promise for a reliable, convenient and closer to the idea of the artificial endocrine pancreas glucose sensor. Subcutaneous glucose levels charted by one of the new sensors were found to be well correlated with venous glucose measurements. Intervention for a healthy lifestyle is frequently hampered by patients' poor compliance. The availability of diverse antidiabetic agents provides options for targeting the glycaemic goal and a choice more fitted to the particularized pathophysiology of each individual subject. Drugs targeting postprandial glycaemia may prove to represent the 'sine qua non' for the 'return' of postprandial glucose values at a 'non-deleterious' threshold, either as monotherapy for the early stages of the disease or as combination therapy later in the progression of diabetes. PMID- 15551343 TI - Size at birth and risk of breast cancer: prospective population-based study. AB - It has been hypothesized that birth size is positively associated with breast cancer risk in adulthood. We studied birth length, birth weight and head circumference at birth and subsequent risk for breast cancer in a cohort of 16,016 women in Norway. Birth length was positively associated with risk (p trend = 002), and women who were 53 cm or longer had a relative risk of 1.8 (CI = 1.2 2.6) compared with women who were shorter than 50 cm, after adjustment for birth year, length of gestation, birth order, maternal age, maternal marital status and socioeconomic status at childbearing. Mutual adjustment for birth weight did not influence the results, and further adjustment for maternal height and adult factors (age at first birth and parity) in a subset of the cohort did not change the results. For birth weight, women in the highest category (>/= 3,840 g) had an adjusted relative risk (RR) of 1.5 (CI = 1.0-2.2) compared to women in the lowest (< 3,040 g), but mutual adjustment for birth length attenuated this association (RR = 1.1; CI = 0.7-1.8). Head circumference at birth showed a similar association as birth weight, with attenuation after mutual adjustment for birth length. The positive association with birth length was stronger among women whose mothers were relatively tall (median or taller, p trend = 0.001) compared to women whose mothers were relatively short (below median, p trend = 0.67) at childbearing. The results provide evidence that intrauterine factors influence future breast cancer risk. The positive association related to birth length suggests that factors that stimulate intrauterine longitudinal growth are particularly important. PMID- 15551344 TI - Interleukin-1B (IL-1B) polymorphisms and gastric mucosal levels of IL-1beta cytokine in Korean patients with gastric cancer. AB - Interleukin-1B and IL-1 receptor antagonist gene polymorphisms are associated with an increased risk of gastric cancer (GC) in Caucasian populations. However, recent studies could not find any association between IL-1B-511T polymorphism and the risk of GC in Asians. We tested for an association between IL-1 loci polymorphisms with increased gastric mucosal levels of IL-1beta and an increased risk of developing GC in a Korean population. Polymorphisms of IL-1A-889, IL-1B 31, IL-1B-511 and IL-1RN were genotyped in 434 controls and 234 patients with GC. Mucosal IL-1beta cytokine was measured using an ELISA. The frequencies of IL-1A, IL-1B-511, IL-1B-31 and IL-1RN were not statistically different between controls and all patients with GC. After subclassification of GC, only patients with intestinal-type GC showed a higher frequency of IL-1B-31T homozygotes (OR = 2.2; 95% CI = 1.1-4.3) compared with controls. Risk was also significantly increased in these patients for IL-1B-31T homozygotes compared with patients with diffuse type GC (OR = 3.4; 95% CI = 1.5-7.7). As in Caucasian populations, linkage disequilibrium between IL-1B-31 and IL-1B-511 was nearly complete, but the pattern of haplotype related to the risk of GC (IL-1B-31T/IL-1B-511C) was opposite (IL-1B-511T/IL-1B-31C). Mucosal IL-1beta levels in H. pylori-infected GC patients were higher in patients homozygous for IL-1B-31T compared with IL-1B 31C/T and IL-1B-31C/C. Thus, the combined effects of H. pylori infection and IL 1B-31T/IL-1B-511C polymorphisms with enhanced mucosal IL-1beta production contributed to the development of intestinal-type GC in this Korean population. PMID- 15551345 TI - Ancient gill and lung oscillators may generate the respiratory rhythm of frogs and rats. AB - Though the mechanics of breathing differ fundamentally between amniotes and "lower" vertebrates, homologous rhythm generators may drive air breathing in all lunged vertebrates. In both frogs and rats, two coupled oscillators, one active during the inspiratory (I) phase and the other active during the preinspiratory (PreI) phase, have been hypothesized to generate the respiratory rhythm. We used opioids to uncouple these oscillators. In the intact rat, complete arrest of the external rhythm by opioid-induced suppression of the putative I oscillator, that is, pre-Botzinger complex (PBC) oscillator, did not arrest the putative PreI oscillator. In the unanesthetized frog, the comparable PreI oscillator, that is, the putative buccal/gill oscillator, was refractory to opioids even though the comparable I oscillator, the putative lung oscillator, was arrested. Studies in en bloc brainstem preparations derived from both juvenile frogs and metamorphic tadpoles confirmed these results and suggested that opioids may play a role in the clustering of lung bursts into episodes. As the frog and rat respiratory circuitry produce functionally equivalent motor outputs during lung inflation, these data argue for a close homology between the frog and rat oscillators. We suggest that the respiratory rhythm of all lunged vertebrates is generated by paired coupled oscillators. These may have originated from the gill and lung oscillators of the earliest air breathers. PMID- 15551347 TI - Nerve conduction study in Sydenham's chorea. AB - Sydenham's chorea (SC) is a late complication of group A beta-hemolytic streptococci infection presumably caused by an abnormal autoimmune reaction. Despite rare case reports of peripheral neuropathy associated with streptococcal infection, there is no investigation of peripheral nerve in SC. We performed nerve conduction studies in a cohort of patients with SC. The neurophysiology investigation comprised measurement of amplitude and sensory conduction velocity of median, ulnar, and sural nerves; amplitude and motor conduction velocity; and F-wave latency of median, ulnar, fibular, and tibial nerves. Twenty-six patients entered the study (12 females, 14 males; mean age 12.8 +/- 3.6 years). Thirteen subjects had absent or decreased deep reflexes. All investigated neurophysiological parameters fell within the normal range for our population. We failed to find neurophysiological evidence of peripheral nerve involvement in patients with a history of SC. Our findings suggest that the possible autoimmune dysfunction in SC patients is not targeted against epitopes present in peripheral nerves. PMID- 15551346 TI - Age-related loss of neuronal nicotinic receptor expression in the aging mouse hippocampus corresponds with cyclooxygenase-2 and PPAR gamma expression and is altered by long-term NS398 administration. AB - Age-related changes in the mammalian dorsal hippocampus are associated with diminished expression of neuronal nicotinic acetylcholine receptors (nAChR), which is particularly severe in pathologies such as those associated with dementias, including Alzheimer's disease. Because the mouse is a useful model for age-related decline in nAChR expression in the basal forebrain and limbic system, we used immunohistochemistry to examine the influence of long-term (12-month) oral administration of nicotine and/or the cyclooxygenase-2 (COX-2) preferring non-steroidal anti-inflammatory drug (NSAID) NS398 on nAChR alpha4, alpha5, alpha7, and beta4 expression in the C57BL/6 mouse. Inhibitory neurons of the dorsal hippocampus that express nAChRs also constitutively express COX-2 and the peroxisome proliferator-antagonist receptor subtype gamma-2 (PPAR gamma2) which is also a target of NS398. Administration of NS398 correlated with retention of nAChR alpha4 and to a lesser extent nAChR beta4, but not nAChR alpha5 or alpha7, but nicotine exhibited no similar effect. Nicotine and NS398 co-administration abolished the NS398-related effect on nAChR alpha4 retention. These results provide evidence that the interaction during aging between oral administration of nicotine and NSAIDs are not straightforward and could even be antagonistic when combined. PMID- 15551348 TI - Levodopa-responsive posttraumatic parkinsonism is not associated with changes of echogenicity of the substantia nigra. PMID- 15551349 TI - Inhibition of peritoneal dissemination of ovarian cancer by tyrosine kinase receptor inhibitor SU6668 (TSU-68). AB - SU6668 (TSU-68) is a small-molecule synthetic inhibitor of the angiogenic related receptor tyrosine kinases Flk-1/KDR, PDGFRbeta, and FGFR1. Using a mouse model of peritoneally disseminated ovarian cancer, we investigated whether SU6668 inhibits peritoneal dissemination and prolongs survival time. BALB/c nude mice were intraperitoneally (i.p.) inoculated with SHIN-3 (VEGF-hypersecretory) or KOC-2S (PDGF-hypersecretory) ovarian serous adenocarcinoma cells with marked peritoneal dissemination ability. From the day after i.p. inoculation of tumor cells, SU6668 was orally administered 6 times weekly at a daily dose of 100 mg/kg or 400 mg/kg. The SU6668-administered group and the vehicle-administered control group were compared for the number of tumor vascular endothelial cells, weight of peritoneally disseminated tumors, amount of ascitic fluid and survival time. As a result, these 3 parameters were significantly smaller in the SHIN-3-inoculated, SU6668-administered mice than in the control group (p = 0.03, p = 0.002, and p = 0.02, respectively). The mean survival time was significantly longer, at 58.1 +/- 11.2 days, in the SU6668-administered mice than that (34.5 +/- 8.8 days) in the control group (p = 0.002). Similarly, in the KOC-2S-inoculated mice, the oral administration of SU6668 significantly reduced these 3 parameters (p = 0.04, p = 0.04, and p = 0.03, respectively), and significantly prolonged survival (16.6 +/- 1.7 days vs. 11.0 +/- 0.7 days, p = 0.008). Thus, the oral administration of SU6668 inhibited angiogenesis and peritoneal dissemination and prolonged survival in mice with peritoneally disseminated ovarian cancer. These effects were observed with both the VEGF- and PDGF-hypersecretory cell lines. Our results suggest that molecular targeting with oral SU6668 will become a new therapeutic strategy targeting peritoneally disseminated ovarian cancer. PMID- 15551350 TI - Novel melanoma antigen, FCRL/FREB, identified by cDNA profile comparison using DNA chip are immunogenic in multiple melanoma patients. AB - We applied a strategy that utilized a combination of systematic gene expression analysis with various tissues and immunological detection with sera from melanoma patients to identify melanoma antigens expressed preferentially in melanoma and melanocytes. We selected 101 genes by comparing cDNA profiles obtained by GeneChip analysis of a highly pigmented melanoma cell line, SKmel23, primary cultured melanocytes, HUVECs cultured endothelial cells, keratinocytes, liver and stomach. After the additional selection with criterion of high registered frequency of each cDNA in melanocyte-related cDNA libraries in the NCBI database, 15 genes including 12 known melanocyte specific genes were identified. One of the remaining 3 genes, FCRL/FREB, encoding a member of the Fc receptor family that was previously reported to express in germinal center B cells, was found to express preferentially in melanocytes and melanoma tissues by RT-PCR and Northern blot analysis. The FCRL/FREB protein was detected in the cytoplasm of melanoma cells by staining with the murine polyclonal antibody and by transfection with GFP-fused FCRL/FREB cDNA. The bacterial recombinant protein was recognized by serum IgG antibody obtained from some patients with melanoma. These results suggest that FCRL/FREB may function in melanocytes and melanoma and may be useful for development of diagnostic methods for various pigment disorders and immunotherapy of melanoma. PMID- 15551351 TI - Gender differences in clinical presentation and response to sertraline treatment of generalized anxiety disorder. AB - OBJECTIVE: To evaluate gender differences in the clinical presentation of generalized anxiety disorder (GAD) and response to sertraline treatment. METHODS: Adult outpatients who met DSM-IV criteria for GAD with a minimum Hamilton rating scale for anxiety (HAM-A) total score>or=18 were randomized to 12 weeks of double blind treatment with flexible doses (50-150 mg) of sertraline (n=182; female, 59%) or placebo (n=188; female, 51%). RESULTS: Clinical presentation of GAD was very similar in men and women in terms of the severity of the HAM-A psychic factor, severity of concomitant depression symptoms, duration of GAD, quality of life and impairment in physical health. Women had an earlier age of onset and higher HAM-A somatic factor scores compared with men. For both men and women, treatment with sertraline resulted in greater change from baseline to endpoint on the HAM-A compared with placebo (adjusted change+/-SE: men:-12.1+/-0.9 vs -8.8+/ 0.9; women: -11.4+/-0.8 vs -7.1+/-0.9, p<0.001); the interaction between gender and treatment group was not significant, nor was there a significant difference between the average change from baseline for men compared with women. Similarly, responder rates based upon clinical global impression-improvement (CGI-I) scores at endpoint showed no significant interaction between gender and treatment, nor was there a significant difference in the response rates by gender; however, the response rate of sertraline compared with placebo was significantly different (p<0.0001) (men: 64% vs 40%; women: 62% vs 34%). Similar findings were evident at week 4 assessment and for completers (week 12). Overall, sertraline was well tolerated by both men and women. DISCUSSION: Women and men with GAD showed similar clinical presentations, with the exception that women had an earlier age of onset and reported more somatic anxiety symptoms. Sertraline was an effective and well tolerated treatment for GAD in both men and women. PMID- 15551352 TI - Potential immunogenicity of adult T cell leukemia cells in vivo. AB - Experimental vaccines targeting human T cell leukemia virus type-I (HTLV-I) Tax have been demonstrated in a rat model of HTLV-I-induced lymphomas. However, the scarcity of HTLV-I-expression and the presence of defective HTLV-I-proviruses in adult T cell leukemia (ATL) cells have raised controversy about the therapeutic potential of HTLV-I-targeted immunotherapy in humans. We investigated the expression of HTLV-I antigens in fresh ATL cells by using both in vitro and in vivo assays. In flow cytometric analysis, we found that 3 of 5 acute-type and six of fifteen chronic-type ATL patients tested showed significant induction of HTLV I Tax and Gag in their ATL cells in a 1-day culture. Concomitantly with HTLV-I expression, these ATL cells expressed co-stimulatory molecules such as CD80, CD86 and OX40, and showed elevated levels of antigenicity against allogeneic T cells and HTLV-I Tax-specific cytotoxic T-lymphocytes (CTL). Representative CTL epitopes restricted by HLA-A2 or A24 were conserved in 4 of 5 acute-type ATL patients tested. Furthermore, spleen T cells from rats, which had been subcutaneously inoculated with formalin-fixed uncultured ATL cells, exhibited a strong interferon gamma-producing helper T cell responses specific for HTLV-I Tax expressing cells. Our study indicated that ATL cells from about half the patients tested readily express HTLV-I antigens including Tax in vitro, and that ATL cells express sufficient amounts of Tax or Tax-induced antigens to evoke specific T cell responses in vivo. PMID- 15551353 TI - Parental occupational exposures and Ewing's sarcoma. AB - A case-control study of Ewing's sarcoma (ES) was conducted to search for occupational exposures associated with ES. The study consisted of 196 cases and 196 random-digit controls matched on geographical region, gender, ethnic origin and birth date. A questionnaire was administered to mothers of participants to obtain information on medical conditions, medications, and parental occupations during and after the index pregnancy. An occupational exposure expert coded jobs and industries for possible and probable exposure to selected occupational hazards. Risk of ES was increased with probable parental exposure to wood dusts during their usual occupation post pregnancy (odds ration [OR] = 3.2; 95% confidence interval [CI] = 1.1-9.2). Other exposures, including a priori suspected risk factors such as exposure to pesticides and farm animals, were not significantly associated with ES. A history of household pesticide extermination was associated with ES among boys aged 15 or younger (OR = 3.0; 95% CI = 1.1 8.1), but not among girls or older boys. Our results suggest that earlier reports of associations of ES with parental farm employment may have been describing risks associated with organic dusts encountered when working on a farm, rather than agricultural exposures or other farming related exposures. PMID- 15551354 TI - Cadmium-induced malignant transformation in rat liver cells: role of aberrant oncogene expression and minimal role of oxidative stress. AB - Our study examined the role of oxidative stress and aberrant gene expression in malignant transformation induced by chronic, low-level cadmium exposure in non tumorigenic rat liver epithelial cell line, TRL 1215. Cells were cultured in 1.0 microM cadmium (as CdCl(2)) for up to 28 weeks and compared to passage-matched control cells. The level of cadmium used for transformation produced no evidence of increased superoxide (O(2) (-*.)) or hydrogen peroxide (H(2)O(2)) levels in the early stages of exposure (6-AN>atrazine approximately copper>ethanol. Results from these studies indicated that these two species responded similarly to the test compounds. Xenopus tropicalis was somewhat less sensitive to 6-AN, semicarbizide and atrazine when tested at 27 degrees C than at 23 degrees C. Ethanol, copper and atrazine were reasonably equipotent in X. tropicalis and X. laevis in terms of teratogenic response (EC50 for malformation), whereas 6-AN and semicarbizide were less potent in X. tropicalis than in X. laevis. No substantial differences (order of magnitude) in potency were observed between X. laevis and X. tropicalis with any of the test materials evaluated. Malformation syndromes induced in both species were similar in X. tropicalis and X. laevis. These results suggested that X. tropicalis could be used effectively as a test organism for the FETAX model. PMID- 15551383 TI - Prevention of fumonisin-induced maternal and developmental toxicity in rats by certain plant extracts. AB - In earlier work we have reported that garlic and cabbage extracts can protect laboratory animals from the toxic effects of different mycotoxins. Previous research demonstrated that fumonisin (FB) induced developmental effects in mice, rats and hamsters. The objectives of the present study were to utilize the pregnant rat as an in vivo model to compare the potential of garlic and cabbage seed extracts to prevent the developmental toxicity of FB and the effects of these extracts on sphingolipid metabolism in dam and foetus livers. Six treatment groups included a control group, a group fed on an FB-containing diet (150 mg kg( 1) feed) and groups treated orally with garlic or cabbage extracts (5 mg kg(-1) body wt.) with or without FB during gestation days 6-15. Evaluations of toxicity were performed on day 20. These include: maternal (mortality, body weight, feed intake and litter weight), developmental (embryonic resorption, foetal body weight, foetal soft-tissue anomalies and foetal skeletal examinations) and maternal and foetal sphingolipid metabolism. Fumonisin alone resulted in significant decreases in feed intake, body weight gain, litter weight, number of live foetuses and foetal body weight, whereas it increased significantly the number of resorbed foetuses and the number of skeletal malformations (30.4% for skull and 26.08% for sternebrae) and also increased the sphinganine/sphingosine (Sa/So) ratio in dam but not fetus livers. Garlic alone or plus FB was comparable to the control regarding all the tested parameters. On the other hand, cabbage seed extract alone or plus FB resulted in 10% maternal mortality and a decrease in maternal body weight and litter weight. It resulted in 4.65% skull malformations in foetuses but it was comparable to the control with regard to the other tested parameters. It could be concluded that both garlic and cabbage seed extracts have protective effects in pregnant rats. Moreover, garlic extract was found to have a greater protective effect than cabbage seed extract. PMID- 15551384 TI - Study of caffeine in urine and saliva of horses subjected to urinary acidification. AB - The study of caffeine in racing horses has been of growing concern in veterinary sports medicine since the Association of Racing Commissioners International (ARCI) stated that it has no valid therapeutic use in racehorses. We examined the kinetic alterations in the urinary excretion and salivary secretion of caffeine in seven horses subjected to urinary acidification using ascorbic acid because this procedure can simulate the acidosis that follows anaerobic exercise. They participated in two treatment groups: the control group (SG) received 500 ml of saline and then 2.0 mg kg(-1) caffeine i.v. 30 min later; and the acidi fi ed group (AG) was subjected to urinary acidification with ascorbic acid at a dose of 0.5 g kg(-1) i.v. and then 2.0 mg kg(-1) caffeine i.v. 30 min later. Samples were collected 30 min before caffeine administration, immediately before caffeine administration (time zero) and at 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 48 and 72 h afterwards. The samples were assayed by gas chromatography. The mean urinary pH for SG was 8.2, but for AG it was as low as 5.9 at 4 h, extending acidosis for up to 8 h. The kinetic curves for the two groups were similar for urinary excretion and salivary secretion. Differences occurred only in peak excretion and peak secretion in SG obtained at 1 h and 30 min, respectively, and in AG at 2 h and 1 h, respectively. This could be explained, in part, to the diuresis in AG compared with SG, resulting in less concentrated urine in the former group. The large difference between the pKa of caffeine and the pH of the medium may be responsible for the similar pharmacokinetics observed for the two groups. PMID- 15551385 TI - A promising peptide antibiotic from Terfezia claveryi aqueous extract against Staphylococcus aureus in vitro. AB - The antimicrobial activity of aqueous and methanolic extracts, as well as partially purified proteins extracted from Terfezia claveryi aqueous extract were investigated against Staphylococcus aureus in vitro. A 5% aqueous extract inhibited the growth of S. aureus by 66.4%, while a methanolic extract was ineffective. Partial protein purification of the aqueous extract using ammonium sulphate precipitation revealed that antimicrobial activity was within the third fraction. This fraction was then subjected to gel filtration using Sephadex G 100. Two peaks were obtained. Peak one possessed higher antimicrobial activity. This peak was then subjected to ion exchange chromatography using DEAE Sephadex. Only peak 4 from the six peaks obtained showed a slight antimicrobial activity. Antimicrobial activities of the aqueous extract and the fractions that showed antimicrobial activity were compared with reference antibiotics. PMID- 15551386 TI - Effect of Manilkara hexandra (Roxb.) Dubard against experimentally-induced gastric ulcers. AB - Effects of the flavonoid rich fraction of the stem bark of Manilkara hexandra (Roxb.) Dubard, have been studied on ethanol, ethanol-indomethacin and pylorus ligated gastric ulcers in experimental animals. Oral administration of the ethyl acetate extract (extract A3) inhibited the formation of gastric lesions induced by ethanol in a dose dependent manner. The protective effect of extract A3 against ethanol induced gastric lesions was not abolished by pretreatment with indomethacin (10 mg kg(-1)). Further, extract A3 inhibited increase in vascular permeability due to ethanol administration. Extent of lipid peroxidation was significantly reduced in animals treated with extract. Extract A3 also inhibited the formation of gastric ulcers induced by pylorus ligation, when administered both orally and intraperitoneally. Moreover, pretreatment with extract A3 increased mucus production and glycoprotein content, which was evident from the rise in mucin activity and TC: PR ratio. PMID- 15551387 TI - Antibacterial potentiality of Mesua ferrea Linn. flowers. AB - The antibacterial efficacy of the methanol extract of whole flowers of Mesua ferrea Linn. was studied against various strains of bacteria. It could inhibit a large number of Gram-positive and Gram-negative bacteria at concentration ranges of 100 to 50 microg/ml, or even lower, as against vibrios and Escherichia coli. In in vivo tests, used at concentrations of 100 and 200 microg/g of body weight, it offered significant protection to Swiss strain of albino mice when challenged with 50 MLD of a virulent strain Sulmonella typhimurium ATCC 6539. Mortality in mice due to these two dosages of the extract alone was insignificant. The extract at 200 microg/g body weight dosage, could significantly reduce the viable count of the strain Sulmonella typhimurium ATCC 6539 in liver, spleen and heart blood of the extract treated challenged mice. PMID- 15551388 TI - A double-blind, placebo-controlled investigation of the effects of two doses of a valerian preparation on the sleep, cognitive and psychomotor function of sleep disturbed older adults. AB - One of the most popular herbal remedies for the alleviation of sleep problems is valerian. However, research into valerian is sparse, and studies differ greatly with respect to design, measures, and preparations used. This clinical study used standardized sleep EEG and psychometric tests to evaluate the clinical efficacy of a valerian preparation (Li 156). A placebo-controlled three way crossover clinical trial was completed using 16 (5 male and 11 female) sleep-disturbed participants (aged 50 to 64 years, mean age 55.9, SD 4.68). Participants slept overnight in a sleep laboratory, following a 21:00 hours dose of valerian 300 mg, valerian 600 mg, or placebo (double-blind). EEG sleep was recorded for each participant at 23:00 hours until 07:00 hours, when a psychometric evaluation was performed the morning after dose. Test periods were separated by six days washout period. Results showed no significant effect between valerian 300 mg, valerian 600 mg or placebo on any EEG parameter or psychometric measure. This suggests valerian at these doses is ineffective as an acute dose for sleep problems. However, valerian is widely used, and is traditionally sedative. Therefore, more research is required into therapeutic dose, types of valerian preparation, and the optimum period of use for therapeutic effect. PMID- 15551389 TI - In vitro anti-tyrosinase activity of 5-(hydroxymethyl)-2-furfural isolated from Dictyophora indusiata. AB - The inhibition of mushroom tyrosinase by methanolic extract of Dictyophora indusiata was evaluated and the bioactive component was characterized and identified as 5-(hydroxymethyl)-2-furfural (HMF) by chromatographic and spectroscopic means. Kinetic studies revealed it to be a noncompetitive inhibitor for the oxidation of L-DOPA. On the basis of these findings some related analogues were also tested for their anti-tyrosinase activity, in order to gain more insight into structure and activity relationship among these heterocyclic compounds. PMID- 15551390 TI - Physical workload, ergonomic problems, and incidence of low back injury: a 7.5 year prospective study of San Francisco transit operators. AB - BACKGROUND: The etiologic role of biomechanical factors for low back injury (LBI) needs to be confirmed in prospective studies that control for psychosocial factors. METHODS: Complete baseline information on 1,233 vehicle operators was gathered during medical examinations and by questionnaire. First LBI during 7.5 years of follow-up was ascertained from insurance records. Hazard ratios and etiologic fractions were analyzed with Cox regression models stratified by injury severity and controlling for age, sex, height, weight, ethnicity, and biomechanical and psychosocial job factors. Severe LBI was defined as medically diagnosed postlaminectomy syndrome, spinal stenosis, herniated lumbar disc, sciatica, or spinal instability. RESULTS: An exponential dose-response relationship was found between weekly driving hours and incidence of first LBI. Indicators of physical workload were more strongly associated with more severe low back injuries compared to less severe injuries. Rates of severe LBI increased 39% for every 10-hr increase in weekly driving (hazard ratio 1.39, 95% confidence interval 1.15-1.68). Higher risks of severe LBI were also found among operators performing heavy physical labor on cable cars (hazard ratio 2.76, 95% confidence intervals 1.24-6.14) or reporting more ergonomic problems at baseline (HR for upper quartile 1.65 (95% confidence interval 1.08-2.50). Estimates of etiologic fractions suggest that reduction of ergonomic problems to the low level currently experienced by 25% of drivers would result in a 19% reduction of severe LBI among all drivers. A change from full- (more than 30 hr) to part-time driving (20-30 hr) could reduce the number of severe LBI by 59%, although this gain would be reduced to 28% at the company level if injuries expected among additional employees, hired to maintain full service are included. CONCLUSIONS: Duration of professional driving and ergonomic problems are independent and preventable risk factors for LBI even after adjustment for psychosocial factors. PMID- 15551391 TI - Effects of occupational solvent exposure on reproductive hormone concentrations and fecundability in men. AB - BACKGROUND: Little is known about the effects of organic solvents on male reproductive health. To assess fertility and reproductive endocrine function in solvent-exposed men, we investigated time-to-pregnancy using a retrospective cohort design and cross-sectionally measured reproductive hormone concentrations in painters and millwrights compared to a reference group of carpenters. METHODS: Detailed occupational, exposure, medical, and time-to-pregnancy histories were obtained by telephone interview. Plasma luteinizing hormone (LH), follicle stimulating hormone (FSH), and testosterone concentrations were determined by immunoassay. Exposure indices, which summarized working life exposure to total solvents, chlorinated solvents, aromatic solvents, and thinners, degreasers, varnishes, and adhesives as a category were calculated from exposure histories. RESULTS: FSH concentrations increased significantly with increasing exposure indices for all solvents and for chlorinated solvents. There were no significant associations of solvent exposure indices with LH or testosterone levels. LH, FSH, and testosterone concentrations also did not differ by job title. Using Cox regression, time-to-pregnancy was non-significantly longer in the painters and millwrights than the carpenters. There was no significant association between time-to-pregnancy and any of the solvent exposure indices; however, it should be noted that some of the pregnancies occurred more than 20 years previously, potentially reducing the reliability of the retrospectively collected pregnancy and exposure data. CONCLUSIONS: The significant associations between FSH levels and solvent exposure indices suggest the potential for adverse effects of solvent exposures on reproductive function in men. PMID- 15551395 TI - Anti-Candida effects of estragole in combination with ketoconazole or amphotericin B. AB - The anti-Candida effects of estragole combined with amphotericin B or ketoconazole, commonly used antifungal drugs for treatment of candidasis, were evaluated in this study. The fractional inhibitory concentration (FIC) indices of both estragole combined with ketoconazole against C. albicans and C. utilis calculated from the checkerboard microtiter assay were 0.28 and 0.50 respectively, indicating significant synergism. These drug combinations exhibited additive effects against C. tropicalis, with FIC index of 0.75. Consistent for the most part with the results from the checkerboard titer tests, the time-kill curves of the tested samples also indicated significant synergism or additive effect between ketoconazole and estragole against the Candida species evaluated. In contrast, amphotericin B showed antagonism in combination with estragole in most experiments of this study. Thus, we have shown the synergistic combination effects between estragole and ketoconazole, which may be effective combinations for the treatment of Candida infections. PMID- 15551394 TI - Further studies on the clinical efficacy of Solanum xanthocarpum and Solanum trilobatum in bronchial asthma. AB - The clinical efficacy of two herbs S. xanthocarpum and S. trilobatum in a dose of 300 mg tds for 3 days was investigated in mild to moderate bronchial asthma. Their effect was compared with standard bronchodilator drugs, salbutamol (4 mg) and deriphylline (200 mg). The respiratory function was assessed by measuring the peak expiratory flow rate (PEFR) using a mini peak flow meter. In addition, improvement in lung function was assessed by physical examination (rhonchi and crepitation) and other symptoms such as cough, breathlessness and sputum. S. xanthocarpum and S. trilobatum produced a progressive improvement in the ventilatory function of asthmatic individuals over 3 days. The scores for rhonchi, cough, breathlessness and sputum were decreased by these drug treatments. The improvement in PEFR and the reduction in other symptom scores clearly indicate a bronchodilator effect, a decrease of oedema and secretions in the airway lumen. The response to these herbs can be considered to be equivalent to that of deriphylline but less than salbutamol. No untoward effects were reported during the study. The present study further confirms the traditional use of these herbs in bronchial asthma. PMID- 15551396 TI - Immunopotentiating effect of a 'Yang'-promoting formula of traditional Chinese medicine on aged female BALB/c mice. AB - The 'Yang'-promoting traditional Chinese medicines (TCM) are used to boost vigor and enhance immunity in humans. In this study, the immunopotentiating effect of VI-28, a 'Yang'-promoting TCM formula containing extracts of radix ginseng, cornu Cervi pantotrichum and radix Salvia miltiorrhizae, was investigated. Groups of 8 month-old female ex-breeder BALB/c mice were fed on ordinary mouse food or food containing a low (0.5%) or high (2%) dose VI-28 for up to 18 weeks. From week 6, mice on the TCM-containing diet were much healthier, stronger and more alert than those on the normal mouse food. Furthermore, their thymuses were significantly bigger and heavier than those of the control mice. Histological examination revealed structural changes typical of thymic involution in mice of the control group, whilst the microstructure of thymuses from mice taking TCM-containing food was comparable to that of mice of a much younger age, indicating a positive effect of VI-28 on slowing down thymic involution. Functional analysis of splenocytes from mice of different groups suggested that oral administration of VI-28 corrected the hyporesponsiveness of T lymphocytes in aged mice. These results have important implications for our understanding of the mechanisms of the immunoboosting effect of TCM. PMID- 15551397 TI - Antioxidant activity of supercritical extract of Melissa officinalis subsp. officinalis and Melissa officinalis subsp. inodora. AB - The antioxidant activity of Melissa officinalis subsp. officinalis and of Melissa officinalis subsp. inodora extracts, obtained by using carbon dioxide under supercritical conditions was investigated. The samples were prepared in two steps. A preliminary extraction at 90 bar and 50 degrees C eliminated the essential oil, then a further extraction at 300 bar and 50 degrees C obtained the high molecular mass extract. These samples were tested for autoxidation and the iron or EDTA-mediated oxidation of linoleic acid at 37 degrees C in the absence of solvent, in in vitro systems. During linoleic acid autoxidation and its EDTA mediated oxidation both M. officinalis and M. inodora extracts showed an antioxidant activity, and no significant differences in their efficacy were observed. None showed any prooxidant activity. PMID- 15551398 TI - Evaluation of the antiherpetic activity of standardized extracts of Achyrocline satureioides. AB - Traditionally, Achyrocline satureioides or 'marcela' has been used in South America for the treatment of several disorders. For the present study, three spray-dried extracts (N1, N2 and N3) were used, all of them prepared with 50% of an hydroethanolic extract rich in flavonoid compounds and 50% of blends of different adjuvants. The cytotoxic concentration which causes destruction in 50% monolayer cells (CC50) was 62.5 microg/ml for the three extracts. The antiviral activity was evaluated by using two different strains of herpes simplex virus (HSV-1) and the best results were obtained with KOS strain and N2 extract. Studies concerning the mechanism of the antiherpetic activity demonstrated that N2 extracts showed no virucidal effect or activity on cellular receptors. HSV-1 DNA synthesis was not inhibited. The antiherpetic activity occurred between the second and ninth hour of the virus replication cycle, probably indicating a perturbation on late stages of this cycle. PMID- 15551399 TI - In vivo and in vitro activities of the seed extract of Piper guineense Schum. and Thonn. against skin and gill monogenean parasites of gold fish (Carassius auratus auratus). AB - Methanol extracts of the seeds of Piper guineense (Piperaceae) were active against gold fish (Carassius auratus auratus L. Pisces Cyprinidae) monogenean parasites. The seed extract of P. guineense was administered at different concentrations (0.5-2.0 mg/L) under in vivo and in vitro conditions. There was a higher efficacy of the effects of the extracts against fish parasites under in vitro situations than under in vivo. Three major compounds (piperanine, N isobutyl (E,E)-2,4 decadienamide and Deltaalpha,beta-dihydrowasanine) were identified from the seed extract of Piper guineense by LC-MS analysis. PMID- 15551403 TI - On the efficiency of targeted clinical trials. AB - The development of genomics-based technologies is demonstrating that many common diseases are heterogeneous collections of molecularly distinct entities. Molecularly targeted therapeutics is often effective only for some subsets patients with a conventionally defined disease. We consider the problem of design of phase III randomized clinical trials for the evaluation of a molecularly targeted treatment when there is an assay predictive of which patients will be more responsive to the experimental treatment than to the control regimen. We compare the conventional randomized clinical trial design to a design based on randomizing only patients predicted to preferentially benefit from the new treatment. Trial designs are compared based on the required number of randomized patients and the expected number of patients screened for randomization eligibility. Relative efficiency depends upon the distribution of treatment effect across patient subsets, prevalence of the subset of patients who respond preferentially to the experimental treatment, and assay performance. PMID- 15551400 TI - Components of diesel exhaust particles differentially affect lung expression of cyclooxygenase-2 related to bacterial endotoxin. AB - We have reported previously that components of diesel exhaust particles (DEP) differently affect acute lung injury related to lipopolysaccharide (LPS) in mice. This study examined the effects of components of DEP on the lung expression of cyclooxygenase (COX)-1 and -2 in the presence or absence of LPS. ICR mice were divided into six experimental groups that received vehicle, LPS (2.5 mg kg(-1)), organic chemicals in DEP (DEP-OC) extracted with dichloromethane (4 mg kg(-1)), residual carbonaceous nuclei after the extraction (washed DEP: 4 mg kg(-1)), DEP OC (4 mg kg(-1)) + LPS (2.5 mg kg(-1)) or washed DEP (4 mg kg(-1)) + LPS (2.5 mg kg(-1)) intratracheally. The expression of mRNA for both COXs in the lung was evaluated 4 h after the intratracheal administration. The magnitude of COX-1 mRNA expression was not altered in each group. The LPS treatment enhanced the COX-2 gene expression compared with vehicle treatment. Washed DEP combined with LPS further increased its expression compared with LPS alone. In contrast, combined treatment of DEP-OC with LPS decreased COX-2 gene expression compared with LPS alone. These results suggest that the residual carbonaceous nuclei of DEP predominantly enhance lung expression of COX-2 rather than the extracted organic chemicals from DEP in the presence of LPS, which is concomitant with the magnitude of acute lung injury in our previous study. PMID- 15551405 TI - Hemoglobin Titusville, a low oxygen affinity variant hemoglobin, in a family of Northern European background. AB - We report the second case of Hb Titusville in a family of Northern European background. This variant hemoglobin caused by an alpha-globin gene mutation has decreased oxygen affinity. Correct diagnosis is clinically important to spare affected individuals extensive investigations into other causes of low oxygen saturation in peripheral blood. PMID- 15551406 TI - Successful umbilical cord blood transplantation for Fanconi anemia using preimplantation genetic diagnosis for HLA-matched donor. AB - Fanconi anemia is a rare autosomal recessive disease characterized by bone marrow failure, developmental anomalies, and a high incidence of myelodysplasia and acute myeloid leukemia. Stem cell transplantation is the only curative treatment. In the absence of matched- sibling donor, an alternative mismatched family or matched unrelated donor can be used, but the results are inferior to the matched sibling transplant and carry a high risk of morbidity and mortality. Preimplantation genetic diagnosis (PGD) has been increasingly used in recent years for mutation analysis for many genetic disorders and results in the birth of healthy children, saving the need for the termination of pregnancy of an affected embryo. The use of PGD for combined analysis of mutation and HLA matching was reported for the first time in 2001. This enables the birth of an unaffected child who can serve as a donor for an affected sibling in need for stem cell transplantation. We report successful cord blood transplantation for a Fanconi anemia patient from his HLA-matched sibling, born after PGD that included mutation analysis for Fanconi anemia and HLA typing. PGD can provide an unaffected donor for a sibling affected by genetic disease in the absence of a compatible related donor. PMID- 15551407 TI - CD5-negative chronic lymphocytic leukemia with indolent clinical course and autoimmune thrombocytopenia, successfully treated with rituximab. AB - A 59-year-old male with lymphocytosis and thrombocytopenia was asymptomatic without lymphadenopathy or hepatosplenomegaly over 10 years. He was admitted to our hospital because his thrombocytopenia had worsened. The clonal lymphocytes appeared as regular small mature lymphocytes on blood films, and bone marrow biopsy showed diffuse infiltration of mature lymphocytes. However, megakaryocytes also presented. The immunophenotypic analysis by flow cytometry revealed that the lymphocytes were positive for CD19, CD20, CD22, and surface membrane immunoglobulin (SmIg) M and D-lambda and were negative for CD5, CD10, CD11c, CD23, and other lineage markers. Expression levels of CD20 and SmIg were strong. The markers were consistent with CD5- CLL with autoimmune thrombocytopenia. He received rituximab, and a rapid decrease of lymphocytes with concomitant increase of platelets was observed. A few cases of CD5- CLL with a stable clinical course have been reported, thought to be B lymphocytosis of undetermined significance (MLUS). This is the first report of CD5- CLL with indolent clinical course associated with autoimmune thrombocytopenia, successfully treated with rituximab. PMID- 15551410 TI - Foodborne infections and intoxications: need of a multidisciplinary approach for control. PMID- 15551411 TI - Criteria for the evaluation of functional foods. Opinion of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG)--(shortened version). PMID- 15551412 TI - Assessing your risk of heart disease. Knowing how doctors calculate the odds can help you to beat them. PMID- 15551413 TI - Bypass linked to better survival rates in high-risk patients. PMID- 15551414 TI - When the temperature's up, cholesterol's down. PMID- 15551415 TI - Why (and how) heart patients should check air quality. PMID- 15551416 TI - How to protect women's hearts. Poll results reveal knowledge gaps; new guidelines offer help. PMID- 15551417 TI - Hopes rise for cell regeneration. Stem cell injections appear to boost heart muscle healing. PMID- 15551418 TI - Home workouts, ample carbs. An active way to watch TV; another round of diet debates. PMID- 15551419 TI - Ask the doctors. My initial blood pressure readings in my doctor's office are as as 50 to 60 points higher than measurements taken later. Even at home, it takes two or so readings before the numbers come down. What do you think? PMID- 15551420 TI - Should elderly people with heart failure conserve their hearts with less strenuous activity, for example by attempting to maintain a low pulse rate while exercising? PMID- 15551422 TI - How much water should children drink each day? PMID- 15551421 TI - My doctor prescribed Cialis [tadalafil] for erectile dysfunction. Anything I need to watch out for? I have a heart condition. PMID- 15551423 TI - Can we get rid of peanut allergens in schools and child care centers? PMID- 15551424 TI - Neighborhood Safety Network. PMID- 15551425 TI - Product recalls. Plush Frog toys sold at Kohl's Department Stores...and Pokemon plush toys. PMID- 15551427 TI - Chest pain? Seek help within minutes. PMID- 15551426 TI - How to beat genes that raise risks. Draw up an accurate family tree, and use it to protect your heart. PMID- 15551428 TI - Fat-reduction surgery doesn't pay off in heart-risk reduction. PMID- 15551430 TI - When heart disease is "silent," sexual problems may send a signal. PMID- 15551429 TI - 4 steps to lower the toll of killer diseases. PMID- 15551431 TI - A mainstay drug underperforms. But a bedside test makes it easier to spot "aspirin resistance". PMID- 15551432 TI - What readers say about statins. Some letters report side effects; here's how doctors respond. PMID- 15551433 TI - Science vs. a fabled heart threat. The fight against a syndrome that triggers sudden death. PMID- 15551434 TI - Ask the doctors. I suffered a heart attack a few years ago, had an emergency angioplasty, and fortunately survived. I've been feeling great. But when I saw my cardiologist recently, he told me that I needed an implantable defibrillator because my heart muscle had been weakened by the heart attack. Do you think I need this? PMID- 15551435 TI - Ask the doctors. My family doctor gave me a clean bill of health, and said my EKG was normal. A few months later, I had a heart attack. How can that happen if I had a normal EKG? PMID- 15551436 TI - Ask the doctors. Can AIDS cause coronary artery disease? PMID- 15551437 TI - [Pills, capsules, and tablets. The new medicines for HIV]. PMID- 15551438 TI - [The Agua Buena Prodefense of Human Rights Association]. PMID- 15551439 TI - [Diabetes, insulin resistance and HIV]. PMID- 15551440 TI - [The CDC, prevention and people living with HIV]. PMID- 15551441 TI - Peripheral neuropathy. Damaged nerves. PMID- 15551442 TI - Health tips. Nosebleeds. PMID- 15551443 TI - Study finds aspirin may prevent certain breast cancers. PMID- 15551444 TI - Vitamin D supplements may reduce risk of falling. PMID- 15551445 TI - Cornea transplants. Restoring sight. PMID- 15551447 TI - Head lice. A common nuisance. PMID- 15551446 TI - Giant cell arteritis. A treatable disorder. PMID- 15551448 TI - I've been experiencing some shortness of breath while doing simple things around the house, like vacuuming or carrying the laundry upstairs. Tests of my lung function turned out normal. Now my doctor wants to check further for possible heart problems. Please explain how shortness of breath can be related to my heart. PMID- 15551449 TI - I was recently diagnosed with a hand deformity my doctor calls Dupuytren's contracture. Can you tell me more about this condition? PMID- 15551450 TI - Athlete's foot is hard to kick. PMID- 15551451 TI - Ultrasound treatment zaps fibroids. PMID- 15551452 TI - Asthmatics: beware bedtime eating. PMID- 15551453 TI - Is organic food healthier? To date, there's no convincing evidence that organic foods are safer or more nutritious than conventionally produced foods. PMID- 15551454 TI - Treating type 2 diabetes. When diet and exercise aren't enough, a broad range of medications can help control our blood sugar. PMID- 15551456 TI - ADHD: not just a childhood disease. PMID- 15551455 TI - Alcohol, drug dependence late in life. Age-associated changes can mask the problem, but the odds of recovery are good. PMID- 15551457 TI - Illnesses linked to suicide risk in elderly. PMID- 15551458 TI - Rheumatoid arthritis treatment progressing. PMID- 15551459 TI - Tips for traveling abroad. PMID- 15551460 TI - Navigating the Medicare drug-card program: help is out there. PMID- 15551461 TI - Over-the-counter not synonymous with safe. PMID- 15551462 TI - Urinary tract infection drug helps pain, hinders cure. PMID- 15551463 TI - Ovarian cancer: not-so-silent. A swollen abdomen, urinary symptoms, and bloating can be warning signs of ovarian cancer, which afflicts 1 in 57 women in the US. PMID- 15551464 TI - Watchful waiting in prostate cancer. PMID- 15551465 TI - Male breast cancer rates rising. PMID- 15551466 TI - Chronic leg ulcers: surgery may prevent recurrence. PMID- 15551467 TI - Is liposuction good for your heart? PMID- 15551468 TI - PAD risk rises with higher lead, cadmium in blood. PMID- 15551469 TI - What's new for warts? PMID- 15551470 TI - Alternative remedies may activate autoimmune disorders. PMID- 15551471 TI - What is a dropped bladder, and how is it treated? PMID- 15551472 TI - What does "enteric-coated" mean? PMID- 15551473 TI - Comorbidity and social factors predicted hospitalization in frail elderly patients. AB - OBJECTIVE: Studies on factors predicting the hospital admission of geriatric patients have reported different findings. The present study was undertaken to examine the rate of hospitalization among a large sample of frail elderly people living in the community and to identify the most important clinical and patient centered factors associated with the hospital admission. STUDY DESIGN AND SETTING: This is an observational cohort study. All patients (n = 1,291) in six Italian home health care agencies were assessed by a trained staff who collected data on the Minimum Data Set for Home Care (MDS-HC) form. We constructed a longitudinal database including MDS-HC data and information on hospital utilization by each patient. RESULTS: During the follow-up of 12 months, the rate of hospitalization was about 26% of the studied sample. Persons living alone were more likely to have a hospital admission than those living with an informal caregiver (odds ratio OR = 2.59, 95% confidence interval CI = 1.82-3.69). Similarly, persons with economic hardship were more frequently hospitalized than those without these problems(OR = 3.01, 95% CI = 1.75-5.18). Comorbidity and previous hospital admission were associated with a higher risk to be hospitalized, too. CONCLUSION: Our results support the hypothesis that a mix of social and health problems are independent predictors of hospitalization. Identification of those factors that best predict hospital admissions and readmissions gives direction for potential interventions and further research toward reducing unnecessary hospitalizations. PMID- 15551475 TI - [57th annual meeting of the Japanese Association for Thoracic Surgery. Sapporo, Japan. October 20-22, 2004. Program and Abstracts]. PMID- 15551474 TI - Lactobacillus paraplantarum sp. now., a new species related to Lactobacillus plantarum. AB - Four strains of facultatively heterofermentative lactobacilli isolated from beer and human feces have physiological characteristics similar to those of Lactobacillus plantarum. Unlike 66% of the L. plantarum strains tested (F. Bringel, M.-C. Curk, and J.-C. Hubert, Int. J. Syst. Bacteriol. 46:588-594, 1996), these strains do not catabolize alpha-methyl-D-mannoside. However, because they exhibit little DNA relatedness to L. plantarum and Lactobacillus pentosus, these four strains were classified as members of a new species, Lactobacillus paraplantarum; strain CNRZ 1885 (= CIP 104668) is the type strain. PMID- 15551476 TI - Tracing the origin of perchlorate. A main ingredient in rocket fuel is showing up almost everywhere researchers look, but where is it all coming from? PMID- 15551477 TI - Ion mobility spectrometers in national defence. PMID- 15551478 TI - Analytical sciences digital library. A Web-based resource for the analytical novice and the seasoned practitioner. PMID- 15551480 TI - Vinland: an inky controversy lives. PMID- 15551479 TI - Do you believe in lipid rafts? Biologists are turning to several analytical techniques to find out whether lipid rafts really exist? PMID- 15551481 TI - Upper processing stages of the perception-action cycle. AB - The neural substrate for behavioral, cognitive and linguistic actions is hierarchically organized in the cortex of the frontal lobe. In their methodologically impeccable study, Koechlin et al. reveal the neural dynamics of the frontal hierarchy in behavioral action. Progressively higher areas control the performance of actions requiring the integration of progressively more complex and temporally dispersed information. The study substantiates the crucial role of the prefrontal cortex in the temporal organization of behavior. PMID- 15551482 TI - Conflict, consciousness, and control. AB - To what degree is executive conflict resolution dependent on conflict awareness? A recent study by Dehaene et al. compared neural responses to conflict elicited through either visible or subliminal primes. Despite behavioral conflict effects for both prime types, neural activity in a control network including the anterior cingulate cortex (ACC) was present only for visible primes. Along with other recent results, these findings have important implications for theories on the relationship between ACC, consciousness, and cognitive control. PMID- 15551483 TI - Binocular battles on multiple fronts. AB - The visual percept fluctuates over time when dissimilar patterns are shown to the two eyes (binocular rivalry). Where in the brain are the corresponding fluctuations of neuronal activity? Conflicting results from studies using various techniques have rendered this question unresolved at present. A recent paper by Wilson, however, promises to reconcile previous disagreements, as it demonstrates that rivalry can be located at different stages of the neuronal hierarchy, depending on the nature of stimulation. PMID- 15551485 TI - Abstracts of the Joint Annual Meeting of the German and Dutch Societies for Immunology (JAMI). 20-23 October 2004, Maastricht, the Netherlands. PMID- 15551484 TI - The neural representation of concrete nouns: what's right and what's left? AB - In a recent paper, Fiebach and Friederici review the literature in the functional imaging of abstract and concrete nouns, and present an fMRI study. They conclude that (a) there is no evidence for a right hemispheric imaginal system that is specifically associated with concrete nouns, and (b) there is evidence supporting aspects of both the context-availability and dual-coding accounts of noun processing. I relate this result to aspects of process and representation, and the anatomy of the ventral temporal lobe. PMID- 15551486 TI - Abstracts of the 148th Meeting of the Acoustical Society of America. San Diego, California, USA. 15-19 November 2004. PMID- 15551487 TI - Dialing for help: state telephone hotlines as vital resources for parents of young children. AB - Toll-free telephone hotlines operated by the states are increasingly being used by families to obtain reliable advice on their young children's health and well being. Originally created for prenatal-care assistance alone, these lines now cover a wide range of early-childhood issues. But while the majority of the lines deliver high-quality information, promptly and empathetically, to their callers, there is still considerable room for improvement. For example, greater use could be made of experts in early-childhood services, and of knowledgeable parents, for speaking with callers and training other staff. The lines could also be made more easily accessible in several ways: through the national 800 number for childhood issues, via the more general 2-1-1 number for community-based services, and by means of a Web site for each line so that it could serve its audience at virtually any time of the day or night. PMID- 15551489 TI - Abstracts of the 5th Central European Orthopaedic Congress. Prague, Czech Republic, July 9-12, 2004. PMID- 15551488 TI - Abstracts of the 9th World Congress on Advances in Oncology and the 7th International Symposium on Molecular Medicine. 14-16 October 2004, Hersonissos, Greece. PMID- 15551490 TI - Abstracts of the 8th Congress of the European Federation for Research in Rehabilitation. Ljubljana, Slovenia, 13-17 June 2004. PMID- 15551491 TI - Carbon nanotubes take new shape. PMID- 15551492 TI - Medicare program; changes to the hospital outpatient prospective payment system and calendar year 2005 payment rates. Final rule with comment period. AB - This final rule with comment period revises the Medicare hospital outpatient prospective payment system to implement applicable statutory requirements and changes arising from our continuing experience with this system and to implement certain related provisions of the Medicare Prescription Drug, Improvement, and Modernization Act (MMA) of 2003. In addition, the final rule with comment period describes final changes to the amounts and factors used to determine the payment rates for Medicare hospital outpatient services paid under the prospective payment system. These changes are applicable to services furnished on or after January 1, 2005. In this final rule with comment period, we are responding to public comments received on the January 6, 2004 interim final rule with comment period relating to MMA provisions that were effective January 1, 2004, and finalizing those policies. Further, we are responding to public comments received on the November 7, 2003 final rule with comment period pertaining to the ambulatory payment classification assignment of Healthcare Common Procedure Coding System (HCPCS) codes identified in Addendum B of that rule with the new interim (NI) comment indicators (formerly referred to as condition codes). PMID- 15551493 TI - Medicare program; revisions to payment policies under the physician fee schedule for calendar year 2005. Final rule with comment period. AB - This final rule refines the resource-based practice expense relative value units (RVUs) and makes other changes to Medicare Part B payment policy. These policy changes concern: supplemental survey data for practice expense; updated geographic practice cost indices for physician work and practice expense; updated malpractice RVUs; revised requirements for supervision of therapy assistants; revised payment rules for low osmolar contrast media; changes to payment policies for physicians and practitioners managing dialysis patients; clarification of care plan oversight requirements; revised requirements for supervision of diagnostic psychological testing services; clarifications to the policies affecting therapy services; revised requirements for assignment of Medicare claims; addition to the list of telehealth services; and, several coding issues. We are making these changes to ensure that our payment systems are updated to reflect changes in medical practice and the relative value of services. This final rule also addresses the following provisions of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (Pub. L. 108-17) (MMA): coverage of an initial preventive physical examination; coverage of cardiovascular (CV) screening blood tests; coverage of diabetes screening tests; incentive payment improvements for physicians in shortage areas; payment for covered outpatient drugs and biologicals; payment for renal dialysis services; coverage of routine costs associated with certain clinical trials of category A devices as defined by the Food and Drug Administration; hospice consultation service; indexing the Part B deductible to inflation; extension of coverage of intravenous immune globulin (IVIG) for the treatment in the home of primary immune deficiency diseases; revisions to reassignment provisions; and, payment for diagnostic mammograms, physicians' services associated with drug administration services and coverage of religious nonmedical health care institution items and services to the beneficiary's home. In addition, this rule updates the codes subject to the physician self-referral prohibition, discusses payment for set-up of portable x ray equipment, discusses the third five-year refinement of work RVUs, and solicits comments on potentially misvalued work RVUs. We are also finalizing the calendar year (CY) 2004 interim RVUs and are issuing interim RVUs for new and revised procedure codes for CY 2005. As required by the statute, we are announcing that the physician fee schedule update for CY 2005 is 1.5 percent, the initial estimate for the sustainable growth rate for CY 2005 is 4.3, and the conversion factor for CY 2005 is $37.8975. PMID- 15551494 TI - Advances in Immunology cumulative subject index, volumes 66-82. PMID- 15551495 TI - Medicare program; prospective payment system for inpatient psychiatric facilities. Final rule. AB - This final rule establishes a prospective payment system for Medicare payment of inpatient hospital services furnished in psychiatric hospitals and psychiatric units of acute care hospitals and critical access hospitals. It implements section 124 of the Medicare, Medicaid, and SCHIP Balanced Budget Refinement Act of 1999 (BBRA). The prospective payment system described in this final rule will replace the reasonable cost-based payment system under which psychiatric hospitals and psychiatric units are paid under Medicare. PMID- 15551496 TI - Revised medical criteria for evaluating malignant neoplastic diseases. Final rules. AB - We are revising the criteria in the Listing of Impairments (the listings) that we use to evaluate claims involving malignant neoplastic diseases. We apply these criteria when you claim benefits based on disability under title II and title XVI of the Social Security Act (the Act). The revisions reflect advances in medical knowledge, treatment, and methods of evaluating malignant neoplastic diseases. PMID- 15551498 TI - Abstracts of the 10th International Congress of Biorheology and the 3rd International Conference of Clinical Hemorheology. Pecs, Hungary, July 18-22, 1999. PMID- 15551497 TI - Dr Leroy's contribution to the understanding of systemic sclerosis. PMID- 15551499 TI - Selling health lifestyles: using social marketing to promote change and prevent disease. AB - As part of its continuing mission to serve trustees and staff of health foundations and corporate giving programs, Grantmakers In Health (GIH) brought together grantmakers, researchers, and public health professionals on May 20, 2004 to discuss the application of social marketing principles to health promotion and chronic disease prevention. As a behavior change technique, social marketing has proven effective in motivating people to make the complex and difficult behavior changes that can improve health and reduce the risk of chronic diseases such as cardiovascular disease, cancer, and diabetes. The Issue Dialogue used the lens of tobacco prevention and cessation, physical activity, and healthy eating to examine how health grantmakers can use social marketing principles and techniques to encourage and support the adoption of healthier behaviors across the lifespan. This Issue Brief incorporates the information and ideas shared at the meeting with a background paper on social marketing that was prepared for participants who attended the Issue Dialogue. It starts with an introduction of social marketing concepts and provides a framework for assessing whether social marketing is an appropriate approach to use in addressing a particular issue. Subsequent sections: (1) describe both the social marketing communications process and techniques, using examples from campaigns developed by health grantmakers and others; (2) describe how social marketing can be used to promote policy change; (3) provide information on communication strategies that can complement social marketing; and (4) present opportunities for grantmakers. PMID- 15551500 TI - Evolution of retroviruses: fossils in our DNA. PMID- 15551501 TI - Circe, Cassandra, and the trojan pigs: xenotransplantation. PMID- 15551502 TI - Donald Sharp Fredrickson. PMID- 15551503 TI - In memory of Dr. Sarwar Jahan Zuberi editor of Pakistan Medical Association (Centre) 1974 - 2004. PMID- 15551504 TI - [Cystic fibrosis. Evidence based respiratory medicine: 2nd update workshop of the SPLF]. PMID- 15551505 TI - [Asthma. Evidence based respiratory medicine: 2nd update workshop of the SPLF]. PMID- 15551506 TI - [Chronic obstructive pulmonary disease exacerbations. Evidence based respiratory medicine: 2nd update workshop of the SPLF]. PMID- 15551507 TI - [Respiratory distress syndrome. Evidence based respiratory medicine: 2nd update workshop of the SPLF]. PMID- 15551508 TI - Think global, act global. PMID- 15551509 TI - From the SNM Committee on Pharmacopeia: Exemption of radiopharmaceuticals from <797>. PMID- 15551510 TI - Exemption of radiopharmaceuticals from <797>. PMID- 15551511 TI - Hypertrophic osteoarthropathy from pulmonary metastatic phyllodes tumour of the breast. PMID- 15551512 TI - Postal questionnaires identified hospitalizations for self-reported acute myocardial infarction. AB - OBJECTIVE: We validated the diagnoses of self-reported acute myocardial infarction (AMI) treated in hospital. STUDY DESIGN AND SETTING: The agreement between myocardial infarction reported in a postal questionnaire in 1998 and data from the Augsburg Coronary Event Register were assessed in a representative sample of German men and women (n = 9,176) aged 25 to 74 years at baseline examination. RESULTS: Of the 9,176 persons, 207 men and women reported an incident AMI treated in hospital during the follow-up period. Of these, 148 persons fulfilled the criteria for verified AMI (positive predictive value 71.5%). Among the 8,969 respondents who reported no AMI, three persons had an AMI (negative predictive value 100%). The sensitivity was 98.0%, and the specificity 99.3%. Much of the false-positive reporting was related to cardiac hospitalizations, predominantly for coronary heart disease (42%). CONCLUSION: A postal questionnaire seems to be a useful method to identify hospitalizations for incident nonfatal AMI cases in epidemiologic cohort studies. Because the proportion of false negatives is low, medical record reviews for case ascertainment can be limited to the group of positive responders. PMID- 15551513 TI - Practical ethics. Keep the chapel? PMID- 15551514 TI - Kaiser targets healthy lifestyles. PMID- 15551515 TI - An overview of assisted living: 2004. PMID- 15551516 TI - Doctors' compensation rose modestly in 2003. PMID- 15551517 TI - Learning collaboratives improve clinics' operations. PMID- 15551518 TI - Terri Schiavo and the law. PMID- 15551519 TI - Squash inhibitors: from structural motifs to macrocyclic knottins. AB - In this article, we will first introduce the squash inhibitor, a well established family of highly potent canonical serine proteinase inhibitors isolated from Cucurbitaceae. The squash inhibitors were among the first discovered proteins with the typical knottin fold shared by numerous peptides extracted from plants, animals and fungi. Knottins contain three knotted disulfide bridges, two of them arranged as a Cystine-Stabilized Beta-sheet motif. In contrast to cyclotides for which no natural linear homolog is known, most squash inhibitors are linear. However, Momordica cochinchinensis Trypsin Inhibitor-I and (MCoTI-I and -II), 34 residue squash inhibitors isolated from seeds of a common Cucurbitaceae from Vietnam, were recently shown to be macrocyclic. In these circular squash inhibitors, a short peptide linker connects residues that correspond to the N- and C-termini in homologous linear squash inhibitors. In this review we present the isolation, characterization, chemical synthesis, and activity of these macrocyclic knottins. The solution structure of MCoTI-II will be compared with topologically similar cyclotides, homologous linear squash inhibitors and other knottins, and potential applications of such scaffolds will be discussed. PMID- 15551520 TI - Conjectures and speculations: Jean Astruc, obstetrics, and biblical criticism in eighteenth-century France. PMID- 15551522 TI - [Medical discourse, hygiene culture, and women in Mexico City at the turn of the century]. PMID- 15551523 TI - Urban trails, human traps: the construction of territories of pleasure and pain in the lives of male homosexuals in the Brazilian northeast in the 1970s and 1980s. PMID- 15551524 TI - Juana "The Mad," the Clares, and the Carthusians: revising a necrophilic legend in early Habsburg Spain. PMID- 15551525 TI - A mysterious success: Doctor Spock and the Netherlands in the 1950s. PMID- 15551526 TI - Discretion and the rule of law: the licensing of drink in England, c. 1817-40. PMID- 15551527 TI - Big hitters tell of their manifesto for Labour. PMID- 15551528 TI - The lack of evidence for ELF magnetic-field effects on bilayer membranes and reconstituted membrane channels. AB - The effects of low-frequency (10-500 Hz) magnetic fields on the electrical properties of channel-free bilayer membranes, and on the single-channel conductance and macroscopic gating characteristics of porin channels incorporated into membranes, have been studied for field strengths in the range 10-100 microT. The field conditions that could in theory give rise to 'cyclotron resonance' effects were also studied. No evidence has been found to support the concept that cyclotron resonance and membrane ion channel effects are involved in the reported biological effects of ELF magnetic fields. PMID- 15551529 TI - The detection and analysis of multiple-membrane-channel events by convolution. AB - A mathematical convolution technique that provides simple, rapid detection of ion transport events when applied to the analysis of biological membrane conductance measurements is described. PMID- 15551530 TI - SAR calculations in an anatomically realistic model of the head for mobile communication transceivers at 900 MHz and 1.8 GHz. AB - A new mathematical model of the head has been constructed from a set of serial MRI slices from one subject. Finite-difference time-domain (FDTD) calculations of the specific energy absorption rate (SAR) have been performed on this model with a 2 mm resolution for a generic mobile communication transceiver represented by a quarter-wavelength monopole on a metal box. The antenna was mounted either at the centre or corner of the top face of the box. The frequencies considered were 900 MHz and 1.8 GHz. Three irradiation geometries were considered, a vertical handset in front of the eye and vertical and horizontal orientations at the side of the ear. The effect of a hand grasping the handset was considered. The head model was scaled to represent the head of an infant and a subset of calculations was performed to verify that the SAR deposited in the infant head did not exceed that in the adult. Results are also presented for a half-wavelength dipole. The maximum SAR values produced by the generic transceiver for the horizontal orientation at the side of the head which is the most typical position, averaged over 10 g of tissue at 900 MHz and 1.8 GHz, are 2.1 and 3.0 W kg(-1) per W of radiated power. The corresponding values over 1 g of tissue are 2.3 and 4.8 W kg( 1) per W at 900 MHz and 1.8 GHz. However, if one were to consider all possible operational conditions, the placement of the transceiver in front of the eye will give 3.1 and 4.6 W kg(-1) per W averaged over 10 g of tissue and 4.7 and 7.7 W kg(-1) per W over 1 g of tissue at 900 MHz and 1.8 GHz, respectively. PMID- 15551531 TI - Absorbed-dose calibrations in high-energy photon beams at the National Physical Laboratory: conversion procedure. AB - The absorbed-dose calibration service from NPL is based on a primary-standard calorimeter that measures absorbed dose to graphite. Secondary-standard dosemeters are calibrated in absorbed dose to water in a 60Co gamma-ray beam and in x-ray beams over a range of generating potentials from 4 MV to 19 MV. Two methods were used to convert the calibrations of working-standard ionization chambers from absorbed dose to graphite into absorbed dose to water. One method involved the use of published interaction data for photons and secondary electrons, and required a knowledge of the chamber construction. The second method involved the calculation of the ratio of absorbed dose in graphite and water phantoms irradiated consecutively in the same photon beam using the photon fluence scaling theorem. The two methods were in agreement to 0.1%. PMID- 15551532 TI - A spin-spin relaxation rate investigation of the gelatin ferrous sulphate NMR dosimeter. AB - Spin-spin NMR relaxation rate in the ferrous sulphate gelatin dosimeter has been studied in terms of pH, gelatin concentration, the addition of benzoic acid, and sample size. It is demonstrated that R2 is more sensitive to changes in Fe3+ ion concentration than R1 when measuring at frequencies of 64 and 100 MHz. pH has an important effect on dose response curves, and oxygen depletion occurs significantly more rapidly in FeSO4 gelatin than in the liquid FeSO4, resulting in a saturation dose of approximately 80 Gy at depths greater than approximately 3 mm in phantom. The concentration of gelatin can be increased to 12% by weight, and the dosimeter will continue to exhibit a linear dose response. Sensitivity is maintained at higher gel concentrations by pH compensation. Addition of low concentration benzoic acid to the system does not alter the dose response of the gelatin FeSO4 system. Finally, spontaneous oxidation of Fe2+ ions does not significantly alter the shape of dose response curves but does result in increases in R2 by up to 4% per day. PMID- 15551533 TI - An investigation of the response of a simple design of plane-parallel chamber. AB - This paper reports the experimental investigation of a simple design of plane parallel electron chamber, which has very thin layers of copper (0.018 or 0.035 mm) as conducting material. Measurements comparing the prototype chambers with other ionization chambers (PTW/Markus, NACP) have been carried out, both in a 60Co gamma-ray beam and in high-energy electron beams. The results show that the Ce factors (proportional to the product of water/air stopping-power ratio and perturbation factor) for converting the in-phantom air-kerma-calibrated chamber reading to the absorbed dose to water are nearly constant for incident electron energies between 4 and 11 MeV for prototype chambers with 0.018 mm thick copper layers and between 4 and 15 MeV for chambers with 0.035 mm thick copper layers. Other aspects concerning these prototype chambers, such as polarity effect, cable effect, collecting efficiency and angular response, have also been studied and the results are presented in this paper. PMID- 15551534 TI - The dosimetric verification of a pencil beam based treatment planning system. AB - A new three-dimensional treatment planning system (TPS) based on convolution/superposition algorithms (TMS-Radix from HELAX AB, Uppsala, Sweden) was recently installed at the University Hospital in Lund. The purpose of the present study was to design a quality assurance and acceptance testing programme to meet the specific characteristics of this convolution model. The model is based on parametrization of a non-measurable quantity-the polyenergetic pencil beam. However, the verification of the treatment planning model is still dependent on numerous comparisons of measured depth-doses and dose profiles. The test programme was divided in two basic parts: (i) model implementation and beam data consistency and (ii) model performance and limitations in special situations. The first part was scheduled for all photon beam qualities available before they could be used for clinical treatment planning. The second part was performed for selected energies only. The results indicate clearly that the model is well suited for clinical three-dimensional dose planning and that the TPS handles data as expected. For example, calculated depth-doses for open and wedge beams at depths larger than the depth of dose maximum and profiles for open beams shows a very good agreement with measurements. However, depth-dose deviations at shallow depths, especially for high energies, were found. Monitor units calculated by the system were accurate for most fields except for very large fields, where deviations of several per cent were found. PMID- 15551535 TI - The quantitative analysis of mammographic densities. AB - Quantitative classification of mammographic parenchyma based on radiological assessment has been shown to provide one of the strongest estimates of the risk of developing breast cancer. Existing classification schemes, however, are limited by coarse category scales. In addition, subjectivity can lead to sizeable interobserver and intraobserver variations. Here, we propose an interactive thresholding technique applied to digitized film-screen mammograms, which assesses the proportion of the mammographic image representing radiographically dense tissue. Observers viewed images on a CRT display and selected grey-level thresholds from which the breast and regions of dense tissue in the breast were identified. The proportion of radiographic density was then calculated from the image histogram. The technique was evaluated for the mammograms of 30 women and is well correlated (R > 0.91, Spearman coefficient) with a six-category subjective classification of radiographic density by radiologists. The technique was found to be very reliable with an intraclass correlation coefficient between observers typically R > 0.9. This technique may have a role in routine mammographic analysis for the purpose of assessing risk categories and as a tool in studies of the etiology of breast cancer, in particular for monitoring changes in breast parenchyma during potential preventive interventions. PMID- 15551536 TI - Cone-beam x-ray microtomography of small specimens. AB - Microtomography is a technique for creating three-dimensional images of the internal structure of objects with high spatial resolution. This can potentially allow inspection of the architecture of breast lumpectomy specimens and visualization of tumours in small animals and also has an application as a tool for non-destructive testing. An efficient method to perform microtomography is to use an area detector and cone-beam reconstruction techniques. In this paper we report on the development of an instrument for microtomography and show example images. The equipment consists of a microfocal x-ray tube (energies and currents up to 30 kVp, 0.2 mA, focal spot size < 5 microm), a rotating specimen stage and a high-resolution x-ray image intensifier optically coupled to a CCD video camera. Data acquisition and 3D image reconstruction are performed by a desktop computer. The well-known Feldkamp cone-beam reconstruction algorithm is used to produce tomographic images from the recorded x-ray projections. The instrument can image samples with diameters of 5-50 mm and create tomographic images with spatial resolution of the order 10-100 microm and signal-to-noise ratio of better than 5:1. This work is a continuation and improvement of an earlier instrument with a low-energy x-ray source and detector. PMID- 15551537 TI - A Monte Carlo study of grid performance in diagnostic radiology: task-dependent optimization for digital imaging. AB - A Monte Carlo computational model has been used to optimize grid design in digital radiography. The optimization strategy involved finding grid designs that, for a constant signal-to-noise ratio, resulted in the lowest mean absorbed dose in the patient. Different examinations were simulated to explore the dependence of the optimal scatter-rejection technique on the imaging situation. A large range of grid designs was studied, including grids with both aluminium and fibre interspaces and covers, and compared to a 20 cm air gap. The results show that the optimal tube potential in each examination does not depend strongly on the scatter-rejection technique. There is a significant dose reduction associated with the use of fibre-interspaced grids, particularly in paediatric radiography. The optimal grid ratio and strip width increase with increasing scattering volume. With increasing strip density, the optimal strip width decreases, and the optimal grid ratio increases. Optimal grid ratios are higher than those used today, particularly for grids with large strip density. It is, however, possible to identify grids of good performance for a range of strip densities and grid ratios provided the strip width is selected accordingly. The computational method has been validated by comparison with measurements with a caesium iodide image receptor. PMID- 15551539 TI - Accelerated EM reconstruction in total-body PET: potential for improving tumour detectability. AB - Total-body positron emission tomography (PET) is a useful diagnostic tool for evaluating malignant disease. However, tumour detection is limited by image artefacts due to the lack of attenuation correction and noise. Attenuation correction may be possible using transmission data acquired after or simultaneously with emission data. Despite the elimination of attenuation artefacts, however, tumour detection is still hampered by noise, which is amplified during image reconstruction by filtered backprojection (FBP). We have investigated, as an alternative to FBP, an accelerated expectation maximization (EM) algorithm for its potential to improve tumour detectability in total-body PET. Signal to noise ratio (SNR), calculated for a tumour with respect to the surrounding background, is used as a figure of merit. A software tumour phantom, with conditions typical of those encountered in a total-body PET study using simultaneous acquisition, is used to optimize and compare various reconstruction approaches. Accelerated EM reconstruction followed by two-dimensional filtering is shown to yield significantly higher SNR than FBP for a range of tumour sizes, concentrations and counting statistics (deltaSNR = 6.3 +/- 3.9, p < 0.001). The methods developed are illustrated by examples derived from physical phantom and patient data. PMID- 15551538 TI - A phantom for quantitative ultrasound of trabecular bone. AB - The propagation mechanisms of ultrasound in trabecular bone are poorly understood and have been the subject of extended debate; also, the reproducibility of ultrasonic measurements on bone in vivo using commercial ultrasound heel-scanning devices is such that the interpretation of the obtained data is difficult. In this paper we describe recent developments in the production of a bone-mimicking material which is well suited to the task of routine monitoring of commercial ultrasound bone scanners. The material, based on a standard epoxy resin is fabricated to a pre-determined porosity value by the inclusion of a marrow mimicking material thereby introducing a known and controlled mean pore size. Measurements of the velocity and attenuation of the material have been performed over a range of porosity values from 10% to 80% in the frequency range 500-900 kHz; also, broadband ultrasonic attenuation (BUA) values have been obtained from commercial equipment. The material displays velocities in the range 1844-3118 m s(-1) and attenuation ranging from 7.0 to 17.7 dB cm(-1) at 500 kHz. PMID- 15551540 TI - Optical-coherence tomography of a dense tissue: statistics of attenuation and backscattering. AB - This paper addresses fundamental issues that underlie the interpretation of images acquired from turbid tissues by optical-coherence tomography (OCT). The attenuation and backscattering properties of freshly excised rat arteries and their dependence on the focusing and collection optics of the OCT system were measured at two wavelengths in the near infrared (830 nm and 1300 nm). Determined from the ratio of the magnitudes of the reflections from glass plates placed on both sides of the arteries, the mean attenuation coefficient of the arterial wall was found to be in the range 14 < microt < 22 mm(-1) at 830 nm and 11 < microt < 20 mm(-1) at 1300 nm. The measured values of microt were lowest for the longer source wavelength and for probe beams with the smallest average diameters. The observed dependence of microt on beam size indicates that relatively large-scale variations in the index of refraction of the tissue contributed to degradation of the tranverse spatial coherence of the beam. We introduce a framework for understanding and quantifying beam-size effects by way of the mutual-coherence function. The fact that spatial variations in backscattering and attenuation (which includes spatial-coherence losses) have similar effects on OCT signals makes the origin of the signals difficult to determine. Evidence is given that suggests that, in spite of this difficulty, certain features of microstructures embedded several hundred micrometres deep in a turbid tissue can still be detected and characterized. PMID- 15551541 TI - Pulsed photothermal radiometry in optically transparent media containing discrete optical absorbers. AB - A description of heat transport by conduction and radiation in inhomogeneous materials following absorption of a brief optical pulse is presented, and investigated experimentally using pulsed photothermal radiometry (PPTR). The model indicates that the role of radiation as an intramedium heat transfer modality increases with increasing temperatures and decreasing infrared (IR) absorption of the medium. However, for the range of conditions analysed in this study, conductive transfer dominates. Thus, the inclusion of radiation does not significantly perturb the internal temperature profiles, although it does influence the radiometric emission from the sample, and hence the PPTR signal. The thermal confinement effects described in this study may be relevant in photomedicine, for example in pulsed laser irradiation of tissues containing small absorbing targets. PMID- 15551542 TI - The Monte Carlo modelling of in vivo x-ray fluorescence measurement of lead in tissue. AB - A Monte Carlo model has been developed, using the EGS4 code, to model the in vivo x-ray fluorescence (XRF) measurement of Pb in non-superficial bone/tissue. Unlike previous work in this field the current model incorporates a correction for Doppler broadening of the Compton scatter peak due to the electron momentum distribution of the medium (tissue/water) in which the photons are Compton scattered by convolving the Compton peak of the Monte Carlo generated spectrum with a modified Compton profile for water. This correction improves the agreement between the measured spectral shape obtained using an experimental in vivo x-ray fluorescence Pb analyser with a 109Cd/180 degrees source/geometry combination, measuring a bone phantom at depth in water and the generated spectral shape obtained from the equivalent Monte Carlo model. The model enables improved estimates to be made of the spectral background beneath the Pb Kalpha1 and Kalpha2 x-ray peaks compared with estimates based on simpler models that assume that Compton interactions are with 'free' electrons and hence permits better optimization of in vivo analyser system design. PMID- 15551543 TI - Inactivation cross section of ions for dry enzymes and viruses. AB - In this note, a semi-empirical analytical formula for the inactivation cross section of ions for dry enzymes and viruses is developed based on the delta-ray theory of track structure. Using the experimental characteristic dose of gamma rays, the cross sections are calculated and compared with experimental results. The average ratio of the experimental to the calculated cross section is 0.96 +/- 0.17. PMID- 15551544 TI - Ultrasonic colour flow imaging. AB - Real-time ultrasonic colour flow imaging, which was first demonstrated to be feasible only about a decade ago, has come into widespread clinical use. Ultrasound is scattered by ensembles of red blood cells. The ultrasonic frequency that gives the best signal-to-noise ratio for backscattering from blood depends on the required penetration. The frequency of ultrasound backscattered from flowing blood is shifted by the Doppler effect. The direction of flow can be determined by phase quadrature detection, and range selectivity can be provided by pulse-echo time-delay measurements. The Doppler frequency spectrum can be determined by Fourier analysis. Early two- and three-dimensional flow-imaging systems used slow manual scanning; velocity colour coding was introduced. Real time colour flow imaging first became feasible when autocorrelation detection was used to extract the Doppler signal. Time-domain processing, which is a broad-band technique, was also soon shown to be practicable, for analysing both radio frequency pulse-echo wavetrains and two-dimensional image speckle. Frequency- and time-domain processing both require effective cancellation of stationary echoes. The time-domain approach seems to have advantages in relation to both aliasing and the effects of attenuation in overlying tissues. Colour-coding schemes that can be interpreted without the need to refer to keys have been adopted, for both velocity and flow disturbance. Colour coding according to signal power has also been reintroduced. Three-dimensional display has been demonstrated. In interpreting colour flow images, it is important to understand the functions of critical system controls and the origins of artifacts. Various strategies can be adopted to increase the image frame rate. The problems of performance measurement and safety need to be kept under review. There are numerous opportunities for further development of ultrasonic colour flow imaging, including improvements in system design, methods of image display, the use of contrast agents and the solution of previously unexplored clinical problems. PMID- 15551545 TI - Dehydration: a model for (low-temperature) argon laser tissue bonding. AB - Despite considerable investigation, the mechanism of laser assisted vascular anastomosis remains unknown. Indications suggest that bonding is the result of thermal action, particularly the thermal denaturation of tissue proteins. However, our own work has led us to conclude that dehydration is an important factor. Hence, we have proposed that laser anastomosis is the result of dehydration at the apposed tissue faces, induced by laser irradiation. This was investigated by comparing the properties of bonds created by dehydration with those created by laser. The bonds were created using parameters consistent with laser anastomoses created in vivo. Results revealed that anastomoses created by dehydration were equivalent to those created by laser, with little difference in strength, histology or response to rehydration. The only significant difference (p < 0.02) was mean bond strength created at temperatures above the denaturation temperature of the tissue (548 g cm(-2) by laser, 994 g cm(-2) by dehydration). Given the similarity of bonds created by the two methods, we conclude that the same mechanism (i.e. dehydration) is likely to be responsible for bonding in both cases and therefore that argon laser bonding is mediated by dehydration. PMID- 15551546 TI - Oxygen diffusion and reaction kinetics in the photodynamic therapy of multicell tumour spheroids. AB - Effects of oxygen diffusion and reaction kinetics in photodynamic therapy are considered in the context of a multicell tumour spheroid model. Steady-state measurements of oxygen made with a Clark-style microelectrode (4 microm diameter tip) enable us to determine the rate of metabolic oxygen consumption and the oxygen diffusion coefficient in 500 microm diameter EMT6/Ro spheroids. These values are 5.77 micromol 1(-1) s(-1) and 1460 microm2 s(-1), respectively. Time dependent electrode measurements of oxygen concentration during laser irradiation of individual Photofrin-sensitized spheroids are fitted to numerical solutions of a pair of diffusion-with-reaction equations. The analysis yields the rate of photodynamic oxygen consumption and a parameter that governs the oxygen sensitivity of photodynamic therapy. These experimentally derived quantities are used to calculate the temporal and spatial distributions of oxygen and the rate of oxygen consumption in a spheroid during irradiation at several fluence rates. The spatial distribution of photodynamic oxygen consumption is strongly fluence rate dependent. Using the experimental and theoretical results developed in this report, previously published survival data are analysed. The analysis indicates that the threshold dose of reacting singlet oxygen in the EMT6/Ro spheroid is 323 +/- 38 micromol 1(-1) (mean +/- SEM). PMID- 15551547 TI - Admittance models for open ended coaxial probes and their place in dielectric spectroscopy. AB - Starting from a rigorous formulation for the admittance of an open ended coaxial probe, this paper identifies the simplifying assumptions that are introduced to obtain more practical solutions. The predictions of the models are compared to each other and to experimental data obtained on an independently calibrated experimental set-up. The advantages and limitations of a model are determined by the theoretical derivation and the numerical solution. The model identified as most suitable for the measurement of the dielectric properties of biological materials is used to determine the relative permittivity and conductivity of muscle and skull bone in the frequency range 1 MHz to 20 GHz. PMID- 15551548 TI - A general solution to charged particle beam flattening using an optimized dual scattering-foil technique, with application to proton therapy beams. AB - This paper describes a dual-scattering-foil technique for flattening of radiotherapeutic charged particle beams. A theory for optimization of shapes and thicknesses of the scattering foils is presented. The result is a universal optimal secondary-scatterer profile, which can be adapted to any charged particle beam by a simple scaling procedure. The calculation of the mean square scattering angle of the beam after passing through the scattering foils is done using the generalized Fermi-Eyges model for charged particle transport. It is shown that the fluence profile in the plane of interest can be made flat to better than 1% inside a predefined beam radius provided the shaped secondary scatterer has the universal radial thickness profile. The thicknesses of the two foils are optimized to minimize the total energy loss. The theory has been tested experimentally in an 180 MeV clinical proton beam. The measured distributions agree well with the calculations. PMID- 15551549 TI - A fundamental study on hyper-thermal neutrons for neutron capture therapy. AB - The utilization of hyper-thermal neutrons, which have an energy spectrum with a Maxwellian distribution at a higher temperature than room temperature (300 K), was studied in order to improve the thermal neutron flux distribution at depth in a living body for neutron capture therapy. Simulation calculations were carried out using a Monte Carlo code 'MCNP-V3' in order to investigate the characteristics of hyper-thermal neutrons, i.e. (i) depth dependence of the neutron energy spectrum, and (ii) depth distribution of the reaction rate in a water phantom for materials with 1/v neutron absorption. It is confirmed that hyper-thermal neutron irradiation can improve the thermal neutron flux distribution in the deeper areas in a living body compared with thermal neutron irradiation. When hyper-thermal neutrons with a 3000 K Maxwellian distribution are incident on a body, the reaction rates of 1/v materials such as 14N, 10B etc are about twice that observed for incident thermal neutrons at 300 K, at a depth of 5 cm. The limit of the treatable depth for tumours having 30 ppm 10B is expected to be about 1.5 cm greater by utilizing hyper-thermal neutrons at 3000 K compared with the incidence of thermal neutrons at 300 K. PMID- 15551550 TI - Optimizing the planning of intensity-modulated radiotherapy. AB - A method of computing optimized intensity-modulated beam profiles has been further developed and used to generate highly conformal radiotherapy dose distributions. The features of these distributions are shown to be strongly dependent on the tuning built into the algorithm. The optimization aims to achieve a specified dose prescription with a posteriori computation of probabilistic biological response. A method has been developed to show the effect of stratifying the intensity-modulated beam profiles into a number of finite intensity increments. It is shown that, provided the number of intensity strata is not too small, highly conformal dose distributions can be achieved with a number of fields (e.g. 7 or 9) which is not excessively large. This number, however, depends on the exact shape of the planning target volume (PTV and its disposition with respect to juxtaposed organs at risk (OARs). These intensity modulated profiles can therefore be delivered either by apparatus for 'tomotherapy' or by using the multileaf collimator at each gantry orientation to deliver a sequence of fixed fields with different field sizes, constructing the beam profile via finite increments of beam intensity. When the PTV and OARS overlap, due to including a finite margin on the clinical target volume to account for tissue movement, it is shown that the dose delivered to the overlap region provides a limit on what can be achieved with conformal therapy. This problem is encountered, for example, when treating the prostate which lies next to part of the rectum and bladder. Some comment is provided on, but not a solution for, the problem of optimizing field orientation. PMID- 15551551 TI - Modelling the effect of charge on selective deposition of particles in a diseased lung using aerosol boli. AB - The technique of using a charged bolus of aerosol to deliver a drug or other agent is advantageous since sites of interest within the lung can be selectively targeted. Ideally, the volume of the bolus should match that of the targeted region allowing the aerosol bolus particles to be confined to the selected area during the pause period after inhalation. Our existing computer model for predicting the deposition of charged aerosol particles has been developed to encompass aerosol boli, some diseased lung morphologies and drug dose administered per breathing cycle. Aerosol deposition in the targeted region is found to be enhanced by increasing particle charge, pause period and particle size. For particles in the size range 1-2.5 microm, aerosol deposition in the region affected by bronchoconstriction does not alter significantly with flow rate variation (range 250-1000 ml s(-1)) for a targeted charged bolus of matched volume. The technique may enable the optimal delivery of therapeutic or other agents to diseased or normal lungs. PMID- 15551552 TI - Improvements in the calibration of 109Cd K x-ray fluorescence systems for measuring bone lead in vivo. AB - A 109Cd K x-ray fluorescence (XRF) system using a point source in a back-scatter geometry is described. The suitability of plaster-of-Paris phantoms as targets for intercalibration standards was evaluated. When the phantom concentrations were measured by inductively-coupled-plasma mass spectrometry (ICPMs), the calculated phantom concentrations underestimated true concentrations by an average of 15%. Since calculated values are used to calibrate the K XRF system, in vivo bone-lead concentrations may be similarly underestimated. The difference between calculated and measured concentration is attributable to impurities in the plaster of Paris (e.g. calcium carbonate). The ICPMS-measured concentrations were used to calibrate the K XRF system. The same phantoms were also measured as 'unknowns' by a bone-lead measurement system (Abiomed, Danvers, MA, USA). The commercial system overestimated the lowest-concentration phantoms and underestimated the phantoms with concentrations above 15 microg Pb/g plaster of Paris. The commercial system and our system were compared by measurement of the new phantoms in air and in water. The K XRF system exhibited better precision in both situations. On the basis of this work, we recommend that plaster-of-Paris phantoms used to calibrate K XRF measurement systems be analysed first by ICPMS or another valid analytical technique. PMID- 15551553 TI - Computer-aided diagnosis in mammography: classification of mass and normal tissue by texture analysis. AB - Computer-aided diagnosis schemes are being developed to assist radiologists in mammographic interpretation. In this study, we investigated whether texture features could be used to distinguish between mass and non-mass regions in clinical mammograms. Forty-five regions of interest (ROIs) containing true masses with various degrees of visibility and 135 ROIs containing normal breast parenchyma were extracted manually from digitized mammograms as case samples. Spatial-grey-level-dependence (SGLD) matrices of each ROI were calculated and eight texture features were calculated from the SGLD matrices. The correlation and class-distance properties of extracted texture features were analysed. Selected texture features were input into a modified decision-tree classification scheme. The performance of the classifier was evaluated for different feature combinations and orders of features on the tree. A classification accuracy of about 89% sensitivity and 76% specificity was obtained for ordered features, sum average, correlation, and energy, during the training procedure. With a leave-one out method, the test result was about 76% sensitivity and 64% specificity. The results of this preliminary study demonstrate the feasibility of using texture information for classification of mass and normal breast tissue, which will be likely to be useful for classifying true and false detections in computer-aided diagnosis programmes. PMID- 15551554 TI - The contrast-detail behaviour of a photostimulable phosphor based computed radiography system. AB - Contrast-detail measurements were performed on a computed radiography imaging system as a function of detector entrance air kerma over the dose range from 0.743 microGy (0.085 mR) to 277 microGy (31.8 mR). A theoretical model of contrast-detail behaviour for a photostimulable phosphor computed radiography system has been derived, which is based on a modified version of the Rose theory of threshold detection. Included in the model are both system and x-ray quantum noise terms, as well as the response of the eye. The zero-frequency noise power of the computed film images was measured with a double-beam scanning microdensitometer. For a given detector dose, good agreement was found between the predicted and measured data when this measurement of system noise was included in the model. The contrast-detail results obtained for the computed radiography system were also compared with contrast-detail results for an image intensifier-TV based digital imaging system and a conventional film-screen system. PMID- 15551555 TI - NMR signal from flowing nuclei in fast gradient-echo pulse sequences with refocusing. AB - A theoretical description of the nuclear magnetic resonance (NMR) signal from flowing nuclei in refocused gradient-echo pulse sequences, both with continuous- and alternating-phase pulse trains, has been developed. Both laminar and plug flow models have been considered and formulae have been derived that relate mean signal intensity to flip angle, pulse sequence repetition interval (TR), and flow velocity. The degree of signal enhancement or reduction in various conditions of flow and pulse sequences depends on the precise phase relationships between the residual transverse magnetization and each radio-frequency (RF) pulse. PMID- 15551556 TI - Position-dependent scatter response functions: will they make a difference in SPECT conducted with homogeneous cylindrical phantoms? AB - This paper explains why it is possible to perform accurate quantitative SPECT when scatter correction is based on stationary and non-stationary scatter functions. This is achieved by comparing the variations of scatter parameters as a function of phantom thickness. The results show that the decrease of scatter fraction with phantom thickness and the decrease of values of scatter kernel inside the field of view are about equal. The deviation of the position-dependent slope from the average value is small for central distributions. These observations explain why estimations of scatter projection by non-stationary convolution and by stationary convolution are comparable when SPECT measurements are conducted with uniform cylindrical phantoms. It is concluded that investigations on the perceived superiority of non-stationary over stationary scatter subtraction in SPECT should be conducted with elliptic phantoms that deviate appreciably from cylindrical shape. PMID- 15551557 TI - A phantom and a procedure to obtain variable-object-contrast test images in gamma camera emission computed tomography (SPECT). AB - The value of test objects with a range of object contrast has been widely recognized for the testing of medical imaging equipment. A simple phantom is described, which, together with a data processing procedure, provides variable contrast rod objects in gamma camera emission computed tomography (SPECT). There is only one compartment to fill with radioisotope, and data from hot rods, cold rods and a uniform section are available for analysis. The rod object contrast varies from 100% to 0%, with nine contrast steps used in this analysis. The measured image contrast in the transaxial slices is shown to be sensitive to variations in the system spatial resolution. With only one compartment to fill with radioisotope, this procedure is applicable for routine checking or optimization of gamma camera SPECT systems. PMID- 15551558 TI - Error estimation of measuring total interaction coefficients of turbid media using collimated light transmission. AB - The error of measuring the total interaction coefficients of turbid media using collimated light transmission was estimated with an analytical expression, which was verified with accurate Monte Carlo simulations. The expression is based on the Henyey-Greenstein phase function of scattering and the probabilities of non scattered and singly scattered photons transmitted through a tissue slab with a unit anisotropy factor. PMID- 15551559 TI - Accuracy of dose calculation methods for retracted tissue compensators. AB - Uncertainties arise in dose calculations involving retracted tissue compensators due to the effects of the compensator upon the scatter component of the dose. Many commercial treatment-planning systems cannot allow directly for the presence of a compensator in isodose calculation or else use simple 2D methods. We present data to test calculation accuracy for a wax compensation system by comparing retraction factors measured along central-axis and off-axis raylines for a variety of compensator shapes, with those derived using effective attenuation coefficient and 3D analytical calculations. The accuracy of using measured uniform-thickness retraction factors for non-uniform shapes and the dose uniformity achievable using a simple compensation system are also discussed. We conclude that the accuracy of simple calculation methods is shape dependent and that calculation errors and dose variations can exceed +/-5% where missing-tissue thickness variations are large. The analytical method is shown to give good agreement with experiment and indicates that it should be possible to adapt algorithms that calculate scatter from the patient for use with compensators. PMID- 15551560 TI - An automatic method for determining the centre of rotation of a mechanically scanned reflection UCT system. AB - A method will be described for determining the centre of rotation of a mechanically scanned reflection ultrasound computed tomography system. It is based on the principle of obtaining opposing images of a test object containing many point targets. The method is automatic in the sense that the centre of rotation is calculated by a computer without the need for an operator to make direct measurements on the mechanical system. For the particular reflection UCT system described here, the centre of rotation is obtained in 3-5 min with a repeatability (+/-2 SD) of +/-0.3 mm. Ways in which even higher accuracy can be obtained are discussed. The basic principle of the method is applicable to any concentric imaging system for which a good approximation to an ideal point target can be produced. PMID- 15551561 TI - Determination of the optical properties of the human uterus using frequency domain photon migration and steady-state techniques. AB - The optical properties (absorption and transport scattering coefficients) of freshly excised, bulk human uterine tissues were measured at 630 nm using frequency-domain and steady-state photon migration techniques. Measurements were made on both normal (pre- and post-menopausal) and non-neoplastic fibrotic tissues. The absorption coefficient of normal post-menopausal tissue (approximately 0.06 mm(-1)) was found to be significantly greater than that of normal pre-menopausal tissue (0.02-0.03 mm(-1)) and pre-menopausal fibrotic tissue (0.008 mm(-1)). The transport scattering coefficient was similar in all three tissue types considered (0.6-0.9 mm(-1)). From the preliminary results presented here, we conclude that optical properties can be reliably calculated either from the frequency-dependent behaviour of diffusely propagating photon density waves or by combining the frequency-independent photon density wave phase velocity with steady-state light penetration depth measurements. Instrument bandwidth and tissue absorption relaxation time ultimately determine the useful frequency range necessary for frequency-domain photon migration (FDPM) measurements. Based on the optical properties measured in this study, we estimate that non-invasive FDPM measurements of normal uterine tissue require modulation frequencies in excess of 350 MHz. PMID- 15551562 TI - Prediction of temperature rise in layered media from measured ultrasonic intensity data. AB - It is now widely recognized that under some circumstances exposure to ultrasound at current diagnostic levels may result in undesirable heating in tissue. The National Council on Radiation Protection and Measurements (NCRP) and the American Institute of Ultrasound in Medicine (AIUM) in conjunction with the National Electrical Manufacturers Association (NEMA) have suggested relatively simple methods for estimating the in situ temperature rise based on simplified ultrasonic intensity profiles. In this paper, the measured spatial intensity distribution from three unscanned ultrasonic transducers is used in an integral solution to the bioheat equation to calculate the temperature increase in layered media, and these calculations are compared with the simple predictions under similar conditions. The transducers included weak and strong focusing devices and one device with a non-cylindrical beam. The ratio between the NCRP and AIUM/NEMA predictions varied from 0.5 to 2.3 in equivalent situations. The ratio between the simple predictions and the more detailed calculations varied from 0.7 to 3.2 for NCRP, and from 0.3 and 3.5 for AIUM/NEMA. These results highlight the need, firstly, to establish simple 'standard' methods for estimating likely temperature increase during clinical examination, and secondly to be clear about whether these methods are intended to give 'worst case' or 'typical' estimates. PMID- 15551563 TI - Dose rate dependence of a PTW diamond detector in the dosimetry of a 6 MV photon beam. AB - The dose rate dependence and current/voltage characteristics of a PTW Riga diamond detector in the dosimetry of a 6 MV photon beam have been investigated. Diamond detectors are radiosensitive resistors whose conductivity (i) varies almost in proportion to dose rate and (ii) is almost independent of bias voltage for a constant dose rate. At the recommended bias of +100 V, and also at +200 V, the detector is operating with incomplete charge collection due to the electron hole recombination time being shorter that the maximum time for an electron to be collected by the anode. As dose rate is varied by changing FSD or depth (changing dose per pulse), detector current and dose rate are related by the expression i alpha Ddelta where delta is approximately 0.98. This manifests itself in an overestimate in percentage depth-dose at a depth of 30 cm of approximately 1% when compared to ionization chamber results. A similar sublinearity is seen when pulse repetition frequency is varied, indicating that the dependence is an on average rather than an instantaneous dose rate. The dose rate dependence is attributed to the reduction in recombination time as dose rate increases. PMID- 15551564 TI - Photon beam quality specification by narrow-beam transmission measurements. AB - Radiation quality specifications in megavoltage photon beams are usually based on depth-dose measurements performed under reference conditions. Stopping-power ratios and various correction factors are then related to parameters such as TPR(10)20, which are extracted from depth-dose measurements. Stopping-power ratio determinations based on this concept were shown to be in error by more than 2% at high energies. Furthermore, electrons generated in the treatment head can, at high energies, contribute to the dose at a depth of 10 cm and thus significantly affect the TPR(10)20 ratio. This method was further shown to be inadequate when the dose in other parts of the field than the reference point was to be measured with ionization chamber dosimetry. A new standardized device for determining photon beam quality based on half value layer (HVL) measurements in water was developed and thoroughly investigated in both a low-energy, (4 MV) and a high energy beam. A relation between HVL and stopping-power ratios water-to-air was determined by comparative measurements with air ionization chambers and liquid filled ionization chambers together with Fricke dosimetry. Furthermore, different radiation quality gradients in the photon fields for different types of field flattening systems, and field-compensating methods were discussed. PMID- 15551565 TI - Estimation of tube potential in mammography from transmission measurements. AB - A computer program has been used to study the performance of a method for estimating the potential of mammographic x-ray tubes. The method uses measurements of the transmission of the x-ray beam through a series of filters. The transmissions are fitted to values calculated using tables of mammographic x ray spectra, thus providing an estimate of tube potential. The performance of the method was studied using a Monte Carlo simulation of experimental errors. The number of measurements and the selection of filter material were both varied, and the precision and accuracy were used as performance indicators. It was found that, for a single filter material, the precision was improved by increasing the number of transmission data, but the accuracy was unaffected. The poorest accuracy and precision were obtained when K-edge filters, or filters with very high or very low atomic number were used. Using combinations of two filter materials, a high transmission (25%) K-edge filter in combination with a low transmission (3%) non-K-edge filter produced results comparable to the best results obtained from seven transmission measurements using a single filter material. By reducing the non-K-edge filter transmission to 1%, an accuracy of +/ 0.4% and a precision of +/- 1.4% were achieved at both 25 kV and 30 kV. PMID- 15551566 TI - Quantitative reconstruction for myocardial perfusion SPECT: an efficient approach by depth-dependent deconvolution and matrix rotation. AB - An efficient reconstruction method for myocardial perfusion single-photon emission computed tomography (SPECT) has been developed which compensates simultaneously for attenuation, scatter, and resolution variation. The scattered photons in the primary-energy-window measurements are approximately removed by subtracting the weighted scatter-energy-window samples. The resolution variation is corrected by deconvolving the subtracted data with the detector-response kernel in frequency space using the depth-dependent frequency relation. The attenuated photons are compensated by recursively tracing the attenuation factors through the object-specific attenuation map. An experimental chest phantom with defects inside myocardium was used to test the method. The attenuation map of the phantom was reconstructed from transmission scans using a flat external source and a high-resolution parallel-hole collimator of a single-detector system. The detector-response kernel was approximated from measurements of a point source in air at several depths from the collimator surface. The emission data were acquired by the same detector setting. A computer simulation using similar protocols as in the experiment was performed. Both the simulation and experiment showed significant improvement in quantification with the proposed method, as compared to the conventional filtered-backprojection technique. The quantitative gain by the additional deconvolution was demonstrated. The computation time was less than 20 min on a HP/730 desktop computer for reconstruction of a 1282 x 64 array from 128 projections of 128 x 64 samples. PMID- 15551568 TI - Absolute quantification of deoxyhaemoglobin concentration in tissue near infrared spectroscopy. AB - We describe a simple technique for non-invasively determining the absolute concentration of deoxyhaemoglobin (Hb) in living tissue using near infrared spectroscopy (NIRS). The technique uses second-differential spectroscopy to determine the relative concentration of Hb to tissue water by fitting of the spectral features of these two chromophores in the 710 nm to 840 nm region of the near infrared (NIR) spectrum. Since the concentration of tissue water is generally known with accuracies of a few per cent, one can then obtain the absolute concentration of Hb ([Hb]). The validity and likely accuracy of the technique is assessed by applying it to artificially generated NIRS data. Some clinical validation is presented by comparing, during an in vivo study, changes in [Hb] obtained by this method and those calculated using more conventional techniques of relative quantification. Finally, we discuss the likely clinical significance of the measurement of absolute Hb concentration. PMID- 15551567 TI - A filtering technique to compensate for detector response in converging-beam SPECT reconstruction. AB - In this study a non-stationary filtering technique was developed to compensate for spatially variant detector response in fan- and cone-beam SPECT. First the frequency-distance relation for fan- and cone-beam geometries was derived from their parallel-beam counterpart, and then used to compute the Fourier transform of the detector response that was used as the MTF of a Metz filter. The filter was applied to the blurred data before reconstruction. The unique feature of the derived MTF is that it depends on source location, so the filtering is spatially variant. Results of evaluation studies demonstrate that the ratio between the tangential and radial FWHMS of the reconstructed point response function obtained from our filtering technique is closer to one and varies less with source location as compared to the ratio obtained without compensation or using Metz filtering with a stationary detector response. This improvement is mainly because the tangential FWHM is increased to match the radial FWHM better. With a higher order of Metz filter, both tangential and radial FWHMS are slightly smaller but their ratio deviates more from one compared to those of lower orders. In conclusion, our filtering technique provides an effective means to compensate for detector response mainly in terms of reducing anisotropy of the point response function. PMID- 15551569 TI - A method of using noise as a test pattern for determining image enhancement filters. AB - Many digitally based medical imaging systems include both reconstruction algorithms and additional image filters designed to enhance certain image features. However, the manufacturers usually consider these algorithms and filters to be proprietory information. The purpose of this note is to describe a simple procedure for determining the spatial frequency response of these proprietary enhancement filters. The technique uses image noise as a test pattern. The procedure consists of acquiring a small number of noise-only data sets (say 10) of a uniform phantom and reconstructing the images using the different filters with repeated use of the noise data sets. A straightforward analysis then yields the enhancement filter frequency responses. PMID- 15551570 TI - Modelling of the dielectric properties of normal and irradiated skin. AB - We have shown that irradiation changes the dielectric properties of human skin at radiofrequencies. Both the dielectric constant and the conductivity of the irradiated skin decrease, especially at low frequencies. The experimental data were analysed using two bioelectric models. Relevant dielectric parameters were determined by curve fitting. The dielectric relaxation of the radiation-induced acute or late reaction of the skin occurred at higher frequencies than with normal or non-irradiated skin, while the static conductivity and static dielectric constant of the irradiated skin decreased. We conclude that bioelectric modelling provides a useful tool in the evaluation of cellular changes in irradiated skin. PMID- 15551571 TI - Reduction of brain noise influence in evoked neuromagnetic source localization using noise spatial correlation. AB - In magnetoencephalographic measurements, magnetic fields caused by spontaneous brain activities not related to the neural activities under study are often referred to as brain noise. This is because the accuracy in neural source localization is considerably degraded by such spontaneous neuromagnetic fields. This paper reports the experimental results of applying the previously proposed noise covariance method to reducing the degradation caused by brain noise and to improving the accuracy in localizing auditory-evoked neural sources. Firstly we present the results of our experiments using measured brain noise and computer generated signal fields. These results confirm that the covariance method can, in principle, improve the accuracy of evoked neural source localization. Next, the method was applied to source localization for actual neuromagnetic fields evoked by speech sounds. The results obtained strongly suggest that the method is effective in processing actual evoked neuromagnetic data. PMID- 15551572 TI - Integrating sphere effect in whole bladder wall photodynamic therapy: I. 532 nm versus 630 nm optical irradiation. AB - The optical absorption, scattering and anisotropy coefficients of piglet bladder, with and without Photofrin, and of diseased human bladder were determined in vitro with a double integrating sphere set-up in the wavelength range 450-800 nm. Monte Carlo simulations were performed in a spherical geometry, representing the bladder, using the optical properties at 532 nm and 630 nm determined in vitro. The calculated fluence rates support the fluence rates that were measured at the bladder wall of a piglet during an in vivo whole bladder wall (WBW) irradiation at 532 nm and 630 nm. Fluence rates calculated and measured in vivo at 630 nm are in agreement with those measured previously in clinical photodynamic therapy (PDT) at 630 nm. WBW-PDT with red light (630 nm) will be technically more advantageous than with green light (532 nm) because of a stronger integrating sphere effect, which reduces the variations of the fluence rate at the bladder wall when the isotropic light source is moved away from the centre of the bladder. Since the optical properties show considerable variations from bladder to bladder, and since as a result the light fluence rate at the bladder wall can vary by a factor of 3 to 4 for the same non-scattered light fluence rate, we conclude that in situ light dosimetry during clinical WBW-PDT is a necessity. PMID- 15551573 TI - Absorbed doses for internal radiotherapy from 22 beta-emitting radionuclides: beta dosimetry of small spheres. AB - We calculated the mean absorbed fractions, specific absorbed fractions and mean doses per unit of cumulated activity in source spheres 10 microm-2 cm in radius for 22 beta-emitting radionuclides potentially useful in radioimmunotherapy. We considered two models of radionuclide distribution, either uniform at the surface of the source or throughout its volume. For each model, we calculated both the absorbed fractions in the spherical segments composing the source and the mean absorbed fractions. For surface distribution, we calculated the mean dose per unit of cumulated activity for a concentric sphere with a small radius (5 microm) in order to determine the minimal dose delivered to the target. Calculations were performed using point kernels for monoenergetic emissions and then integrated into the beta spectra of the different emitters (32p, 33p, 47Sc, 67Cu, 77As, 90Y, 105Rh, 109Pd, 111Ag, 121Sn, 131I, 142Pr, 143Pr, 149Pm, 153Sm, 159Gd, 166Ho, 177Lu, 186Re, 188Re, 194Ir and 199Au). Monoenergetic emissions were taken into account. Results are reported in the form of tables to facilitate use during dosimetric studies for radioimmunotherapy. An application is presented showing the potential utility of associating emitters with different energies in order to sterilize a range of tumour targets of variable size. PMID- 15551574 TI - Energy loss of 70 MeV protons in tissue-substitute materials. AB - Measurements of the energy loss of 70 MeV protons in phantom materials have been made, with a small uncertainty. Average stopping powers relative to water have been determined for the energy range between approximately 70 MeV and 40 MeV. These are expressed in terms of the mean excitation energy I of the Bethe theory. The effect of grains of inhomogeneous materials (e.g. calcium fluoride) has been explored and understood. PMID- 15551575 TI - Methods for fully automated verification of patient set-up in external beam radiotherapy with polygon shaped fields. AB - Methods have been developed to perform fully automated comparisons of the radiotherapy simulator and portal images in the most common cases where the field edges consist of straight lines. The field defining wires or rods in the simulator image, and the edges of the portal image, are localized by means of a truncated Radon transform. Edges are enhanced with the use of a generalized Laplacian operator. From the detected field outlines, a geometrical mapping function is determined that rotates, scales, and translates one image with respect to the other. A subsequent match of field shapes is executed. If satisfactory agreement is established, the anatomical structures of the simulator and portal images are compared by correlating a new pair of images that only contain the intensity ridges representing bone outlines. The correlation determines the rotation and translation that must be applied to align the anatomical structures. With the simulator image already processed, the remaining automatic processing takes approximately 2 min on a SUN Sparcstation 2. By use of a priori knowledge of the fields, the computation time needed after acquisition of the portal image may be reduced to about 40 s for 512 x 512 images and about 12 s for 256 x 256 images. PMID- 15551576 TI - The dependence of ultrasonic properties on orientation in human vertebral bone. AB - Speed of sound (SOS) and broad-band ultrasound attenuation (BUA) were measured in cubes of human trabecular bone from lumbar vertebrae, in the three major anatomical axes. There were significant differences in sos and in BUA when measured in the different axes, indicating a structural component to the ultrasonic measurement. Qualitatively different behaviour was observed in the cranio-caudal (CC) axis compared to the transverse directions: SOS was approximately 500 m s(-1) greater than in either the lateral (LT) or antero posterior (AP) axes, and BUA was approximately 23 dB MHz(-1) cm(-1) greater. Small, but significant, differences existed between the AP and LT axes for both SOS and BUA. In the AP and LT directions, strong linear correlations existed between sos and apparent density (r = 0.90), and between BUA and apparent density (r = 0.96). In the cc axis, correlations with density were poorer. The anomalous behaviour in the cc axis was due to a transient travelling ahead of the main wavefront, and it is suggested that this represents propagation of ultrasound directly through the trabecular framework as a bar wave. This can only occur in the cc axis where the majority of trabeculae are orientated parallel to the direction of propagation. Measurements on cubes in air, as opposed to water, supported this hypothesis. Modifications to the experimental technique necessary to consistently detect this phenomenon are described. PMID- 15551577 TI - A new phase function approximating to Mie scattering for radiative transport equations. AB - A new scattering phase function is presented to approximate the accurate phase function from Mie theory. The new phase function demonstrates much better agreement with Mie theory for biomedical media (with the mean cosine of scattering angles g > 0.8) than the Henyey-Greenstein phase function, which has been widely adopted by researchers in the biomedical optics field. Using the new phase function as the integral kernel in the radiative transport equation, an analytical result for the integral term of the equation is obtained for highly aligned beams. This may lead to a semi-analytical solution to the time-dependent radiative transport equation for time-resolved spectroscopy. PMID- 15551578 TI - Ion loss due to initial recombination in a parallel-plate cavity ionization chamber. AB - Collection efficiencies for ion loss due to initial recombination and back diffusion were measured for several humidities using a parallel-plate cavity ionization chamber irradiated with 60Co gamma-rays. It was shown, from measurements in a range of inverse electric field strengths from 0.05 to 14 mm V( 1), that initial recombination took place both in clusters and columns of ions produced along the path of the secondary electrons ejected by the y-rays. The ion loss due to recombination in clusters was found to increase with humidity, but that in columns did not. Effects of ion clustering reactions on recombination may be reduced after longer periods of ion drift, when recombination in columns takes place. Ion loss due to back-diffusion was also found to have no dependence on humidity. PMID- 15551579 TI - The effects of humidity on volume recombination in ionization chambers. AB - Saturation curves were measured with a parallel-plate ionization chamber for air with humidities of 0%-81%. From the curves, values of m--which is equal to (alpha/eK+ K-)1/2 where alpha is the recombination coefficient, e the charge of an ion, and K+ and K- are mobilities of positive and negative ions, respectively were obtained. It was confirmed that humidity was the decisive cause of the increase in the value of m with the lifetime of ions in the ionization chamber. It was found that the value of m increases sharply in a range of small values of the product of the ion lifetime and the partial pressure of water vapour and increases more slowly at high values of this parameter. PMID- 15551580 TI - Radiance and particle fluence. AB - The concepts of radiance and fluence are fundamental to the description of a radiation field. The International Commission on Radiological Units and Measurements (ICRU) has defined fluence in terms of the number of the radiation particles crossing a small sampling sphere. A second definition has been proposed in which the length of track segments contained within any sampling volume are used to calculate the incident fluence. This approach is often used in Monte Carlo simulations of individual particle tracks, allowing the fluence to be scored in small volumes of any shape. In this paper we stress that the second definition generalizes the classical (ICRU) concept of fluence. We also identify the assumptions inherent in the two definitions of fluence and prove their equivalence for the case of straight-line particle trajectories. PMID- 15551581 TI - Determination of the relative linear collision stopping power and linear scattering power of electron bolus material. AB - The linear collision stopping power and linear scattering power for machineable wax relative to water have been determined for electron energies between 2 and 20 MeV. Knowledge of these quantities is necessary for the use of this wax as bolus in electron pencil-beam dose algorithms. The atomic composition of the wax (rho = 0.920 +/- 0.001 g cm(-3)) was obtained by having the wax assayed. The formalisms expressed in the ICRU Report 35 were used to calculate the relative linear collision stopping and linear scattering powers of the wax. The calculated relative linear collision stopping powers of 2 to 20 MeV electrons in the wax ranged from 0.949 +/- 0.005 to 0.952 +/- 0.005, and the calculated relative linear scattering powers ranged from 0.734 +/- 0.004 to 0.729 +/- 0.004. As a check of the calculation method, the relative linear collision stopping power was measured by determining the shift in electron central-axis depth-ionization curves when varying thicknesses of water were replaced by wax. These measurements, made using 10, 12, 15 and 18 MeV electron beams with wax thicknesses from 1.0 - 4.0 cm, resulted in a mean value of 0.931 +/- 0.008. Determination of the relative linear stopping power and the linear scattering power by using the measured CT number to extract values from patient data tables resulted in values of 0.933 +/- 0.009 and 0.746 +/- 0.016, respectively, indicating that it should be acceptable to use the Hounsfield values obtained with CT scans for treatment planning dose calculations. PMID- 15551582 TI - A performance study of 3D reconstruction algorithms for positron emission tomography. AB - This paper investigates the statistical and systematic accuracy of five three dimensional reconstruction algorithms for multi-ring PET scanners operated without septa: the reprojection method, the direct Fourier reconstruction, the FAVOR algorithm, and the single-slice and multi-slice rebinning algorithms. Simulated data of a uniform cylinder, of Gaussian sources, and of spherical sources are used to compare respectively the noise properties, the modulation transfer function, and the recovery coefficients of the algorithms. Brain scans reconstructed with the different algorithms are compared by calculating the linear regression of the mean values within regions of interest. The most significant observations are a slight loss of transaxial resolution with the reprojection algorithm in the external slices of the scanner, and increased noise in the images reconstructed using multi-slice rebinning. PMID- 15551583 TI - Three-dimensional image reconstruction for PET by multi-slice rebinning and axial image filtering. AB - A fast method is described for reconstructing volume images from three dimensional (3D) coincidence data in positron emission tomography (PET). The reconstruction method makes use of all coincidence data acquired by high sensitivity PET systems that do not have inter-slice absorbers (septa) to restrict the axial acceptance angle. The reconstruction method requires only a small amount of storage and computation, making it well suited for dynamic and whole-body studies. The method consists of three steps: (i) rebinning of coincidence data into a stack of 2D sinograms; (ii) slice-by-slice reconstruction of the sinogram associated with each slice to produce a preliminary 3D image having strong blurring in the axial (z) direction, but with different blurring at different z positions; and (iii) spatially variant filtering of the 3D image in the axial direction (i.e. 1D filtering in z for each x-y column) to produce the final image. The first step involves a new form of the rebinning operation in which multiple sinograms are incremented for each oblique coincidence line (multi slice rebinning). The axial filtering step is formulated and implemented using the singular value decomposition (SVD). The method has been applied successfully to simulated data and to measured data for different kinds of phantom (multiple point sources, multiple discs, a cylinder with cold spheres, and a 3D brain phantom). PMID- 15551584 TI - A methodology for testing for statistically significant differences between fully 3D PET reconstruction algorithms. AB - We present a practical methodology for evaluating 3D PET reconstruction methods. It includes generation of random samples from a statistically described ensemble of 3D images resembling those to which PET would be applied in a medical situation, generation of corresponding projection data with noise and detector point spread function simulating those of a 3D PET scanner, assignment of figures of merit appropriate for the intended medical applications, optimization of the reconstruction algorithms on a training set of data, and statistical testing of the validity of hypotheses that say that two reconstruction algorithms perform equally well (from the point of view of a particular figure of merit) as compared to the alternative hypotheses that say that one of the algorithms outperforms the other. Although the methodology was developed with the 3D PET in mind, it can be used, with minor changes, for other 3D data collection methods, such as fully 3D cr or SPECT. PMID- 15551585 TI - Evaluation of task-oriented performance of several fully 3D PET reconstruction algorithms. AB - The relative performance of five fully 3D PET reconstruction algorithms is evaluated. The algorithms are a filtered backprojection (FBP) method and two variants each of the EM-ML and ART iterative methods. For each of the iterative methods, one variant makes use of voxels and the other makes use of 'blobs' (spherically symmetric functions smoothly decaying to zero at their boundaries) as basis functions in its discrete reconstruction model. The methods are evaluated from the point of view of the efficacy of the reconstructions produced by them for three typical medical tasks--estimation of the average activity inside specific regions of interest, detection of hot spots, and detection of cold spots. A free parameter is allowed in the description of each of the five algorithms; the parameters are determined by a training process during which a value of the free parameter is selected which (nearly) maximizes a technical figure of merit. Such training and the actual comparative evaluation is done by making use of randomly generated phantoms and their projection data. The methodology allows assignation of levels of statistical significance to claims of the relative superiority of one algorithm over another for a particular task. We find that using blobs as basis functions in the iterative algorithms is definitely advantageous over using voxels. This result has high statistical significance. (We also include a visual illustration of it.) Comparing FBP, EM-ML using blobs, and ART using blobs, we do not find a clear difference in the overall performance of the investigated variants of the methods. If anything, our results suggest that ART using blobs may be the most efficacious of the three. PMID- 15551586 TI - The performance of the single-slice rebinning technique for imaging the human striatum as evaluated by phantom studies. AB - The single-slice rebinning (SSRB) technique has been implemented on a ring-based tomograph with removable septa and its performance evaluated with phantom studies. The studies involved the determination of the plane efficiency profile and the mapping of the point spread function. The image quality obtained using the SSRB technique has been compared to that obtained using the standard 3D reconstruction method in terms of the following figures of merit: recovery as a function of source axial position and slice averaging, contrast recovery and contrast to background noise ratio as a function of source axial position. The effect of the region of interest (ROI) size on such a comparison has been evaluated. Finally the effects of different contrast conditions and ROI position on a striatal phantom image have been thoroughly explored. PMID- 15551587 TI - Oversampled filters for quantitative volumetric PET reconstruction. AB - Three-dimensional filtered backprojection uses filters generally specified in the Fourier domain. Implementing these filters by direct sampling in the Fourier domain produces an artifact in the reconstructed images consisting primarily of a DC shift. This artifact is caused by aliasing of the reconstruction filter. We have developed a filter construction technique using Fourier domain oversampling, which reduces the artifact. A method to construct the filter efficiently without the need to create and store the entire oversampled filter array is also presented. Quantitative accuracy in filtered backprojection is of particular importance in multiple-pass algorithms used to reconstruct data from cylindrical PET scanners. We are able to implement such algorithms without fitting the reprojected views to the scanner data. PMID- 15551588 TI - Simulation studies on a pseudo-three-dimensional reconstruction algorithm for volume imaging in multi-ring PET. AB - The imaging performance of a pseudo-three-dimensional (3D) reconstruction algorithm has been investigated. The algorithm is based on ID filtering followed by 3D backprojection. High- and low-frequency components of the image are reconstructed separately, and oblique projections are used only for reconstructing the high-frequency component. The cross talk between different slices due to oblique backprojection is eliminated by high-pass filtering. 'Variable filters', the cut-off frequencies of which are proportional to the tilt angles of projections, provide higher signal-to-noise ratio than 'fixed filters' with similar capability for cross-talk rejection. The maximum ring difference used in the reconstruction of each slice is extended as far as possible, while still satisfying the requirement that the total density of backprojection beams deposited in the slice is approximately uniform in the field of view. This allows the use of almost all available projection data to improve signal-to-noise ratio in peripheral slices. The root mean square noise of the reconstructed images evaluated with simulation studies is in good agreement with numerical calculations, and it is shown that the loss in signal-to-noise ratio caused by discarding the low-frequency component of oblique projections is negligibly small as long as suitable variable filters are used. PMID- 15551589 TI - Rotating positron tomographs revisited. AB - We have compared the performance of a PET scanner comprising two rotating arrays of detectors with that of the more conventional stationary-ring design. The same total number of detectors was used in each, and neither scanner had septa. For brain imaging, we find that the noise-equivalent count rate is greater for the rotating arrays by a factor of two. Rotating arrays have a sensitivity profile that peaks in the centre of the field of view, both axially and transaxially. In the transaxial plane, this effect offsets to a certain extent the decrease in the number of photons detected towards the centre of the brain due to self absorption. We have also compared the performance of a rotating scanner to that of a full-ring scanner with the same number of rings. We find that a full-ring scanner with an axial extent of 16.2 cm (24 rings) is a factor of 3.5 more sensitive than a rotating scanner with 40% of the detectors and the same axial extent. PMID- 15551590 TI - A convolution-subtraction scatter correction method for 3D PET. AB - 3D acquisition and reconstruction in positron emission tomography (PET) produce data with improved signal-to-noise ratios compared with conventional 2D slice oriented methods. However, the sensitivity increase is accompanied by an increase in the number of scattered photons and random coincidences detected. This paper presents a scatter correction technique for 3D PET data where an estimate of the scattered photon distribution is subtracted from the data before reconstruction. The scatter distribution is estimated by iteratively convolving the photopeak projections with a mono-exponential kernel. The method accounts for the 3D acquisition geometry and nature of scatter by performing the scatter estimation on 2D projections. The assumptions of the method have been investigated by measuring the variation in the scatter fraction and the scatter function at different positions in a cylinder. Both parameters were found to vary by up to 50% from the centre to the edge of a large water-filled cylinder. Despite this, in a uniform cylinder containing water with different concentrations of radioactivity, scatter was reduced from 25% in a non-radioactive region to less than 5% using the convolution-subtraction method. In addition, the relative concentration of a cylinder containing an increased concentration, which was underestimated by almost 50% without scatter correction, was within 5% of the true concentration after correction. PMID- 15551592 TI - The cardiofocal collimator: a variable-focus collimator for cardiac SPECT. AB - Investigators in nuclear medicine have long been in search of a practical method to increase the number of detected events in cardiac SPECT. A clinically practical method requires a simple data acquisition protocol, clinically acceptable reconstruction times, artifact levels near or below visual threshold, and the use of currently available cameras and computers. Towards this end, we have developed the Cardiofocal collimator, a variable-focus collimator for cardiac SPECT that increases the number of detected events from the heart by more than a factor of two compared to that of a parallel-hole collimator with equivalent resolution. In both the transverse and axial dimensions, the focusing is strongest at the centre of the collimator, and gradually relaxes to nearly parallel-hole collimation at the edge of the collimator. The variable-focus concept provides an increase in the number of counts from organs imaged near the centre of the collimator, where the heart will spend most of the time during a cardiac SPECT study, while adequately sampling enough of the background activity distribution to prevent truncation artifacts in the reconstructed images. Images are reconstructed in clinically acceptable times using a filtered backprojection reconstruction algorithm. The algorithm supports both full-scan (360 degrees) and short-scan (180 degrees plus the fan angle) acquisitions. The results of simulations and phantom studies are included to demonstrate the performance of the Cardiofocal collimator. PMID- 15551591 TI - Pinhole collimation for ultra-high-resolution, small-field-of-view SPECT. AB - The objective of this investigation was to evaluate small-field-of-view, ultra high-resolution pinhole collimation for a rotating-camera SPECT system that could be used to image small laboratory animals. Pinhole collimation offers distinct advantages over conventional parallel-hole collimation when used to image small objects. Since geometric sensitivity increases markedly for points close to the pinhole, small-diameter and high-magnification pinhole geometries may be useful for selected imaging tasks when used with large-field-of-view scintillation cameras. The use of large magnifications can minimize the loss of system resolution caused by the intrinsic resolution of the scintillation camera. A pinhole collimator has been designed and built that can be mounted on one of the scintillation cameras of a triple-head SPECT system. Three pinhole inserts with approximate aperture diameters of 0.6, 1.2 and 2.0 mm have been built and can be mounted individually on the collimator housing. When a ramp filter is used with a three-dimensional (3D) filtered backprojection (FBP) algorithm, the three apertures have in-plane SPECT spatial resolutions (FWHM) at 4 cm of 1.5, 1.9 and 2.8 mm, respectively. In-air point source sensitivities at 4 cm from the apertures are 0.9, 2.6 and 5.7 counts s(-1) microCi(-1) (24, 70 and 154 counts s( 1) MBq(-1)) for the 0.6, 1.2 and 2.0 mm apertures, respectively. In vitro image quality was evaluated with a micro-cold-rod phantom and a micro-Defrise phantom using both the 3D FBP algorithm and a 3D maximum likelihood-expectation maximization (ML-EM) algorithm. In vivo image quality was evaluated using two (315 and 325 g) rats. Ultra-high-resolution pinhole SPECT is an inexpensive and simple approach for imaging small animals that can be used with existing rotating camera SPECT system. PMID- 15551593 TI - Three-dimensional reconstruction from cone-beam data in O(N3 log N) time. AB - We have used direct Fourier techniques to modify and implement the 3D reconstruction method from cone-beam projections proposed by Grangeat. In this way we manage to decrease the computational complexity from O(N4) to O(N3 log N). Just as Grangeat's original method is exact in the mathematical sense, so is our method, provided a complete set of projection data is acquired. Also in accordance with Grangeat, our algorithm consists of two distinct phases: phase 1, from cone-beam data to derivatives of Radon data; phase 2, from derivatives of Radon data to reconstructed 3D object. In phase 1 we use the direct Fourier method in reverse to obtain line integrals in the detector plane. In phase 2 the 2D linogram method is employed for reconstruction of vertical and horizontal planes in the Radon space. PMID- 15551594 TI - Implementation of Tuy's cone-beam inversion formula. AB - Tuy's cone-beam inversion formula was modified to develop a cone-beam reconstruction algorithm. The algorithm was implemented for a cone-beam vertex orbit consisting of a circle and two orthogonal lines. This orbit geometry satisfies the cone-beam data sufficiency condition and is easy to implement on commercial single photon emission computed tomography (PECT) systems. The algorithm, which consists of two derivative steps, one rebinning step, and one three-dimensional backprojection step, was verified by computer simulations and by reconstructing physical phantom data collected on a clinical SPECT system. The proposed algorithm gives equivalent results and is as efficient as other analytical cone-beam reconstruction algorithms. PMID- 15551595 TI - The importance and implementation of accurate 3D compensation methods for quantitative SPECT. AB - The purpose of this study was to investigate the importance of 2D versus 3D compensation methods in SPECT. The compensation methods included in the study addressed two important degrading factors, namely attenuating and collimator detector response in SPET. They can be divided into two general categories. The conventional methods are based on the filtered backprojection algorithm, the Chang algorithm for attenuation compensation and the Metz filter for detector response compensation. These methods, which were computationally efficient, could only achieve approximate compensation due to the assumptions made. The quantitative compensation methods provide accurate compensation by modelling the degrading effects at the expense of large computational requirements. Both types of compensation methods were implemented in 2D and 3D reconstructions. The 2D and 3D reconstruction/compensation methods were evaluated using data from simulation of brain and heart, and patient thallium SPECT studies. Our results demonstrate the importance of compensation methods in improving the quality and quantitative accuracy of SPECT images and the relative effectiveness of the different 2D and 3D reconstruction/compensation methods. We concluded that 3D implementation of the quantitative compensation methods provides the best SPECT image in terms of quantitative accuracy, spatial resolution, and noise at a cost of high computational requirements. PMID- 15551596 TI - Generalized dual-energy-window scatter compensation in spatially varying media for SPECT. AB - The detection of scattered photons in the photopeak energy window hinders accurate activity estimation in single-photon-emission computed tomography (SPECT). To compensate for photons scattered in spatially varying media, a framework for generalized dual-energy-window scatter subtraction has been developed. Generalized scatter subtraction factors are introduced, and these factors are decomposed into terms dependent on the uniform (average) and spatially varying components of the source activity distribution. The variation of these factors with projection pixel location and gamma camera position is analysed for a simulated myocardial perfusion study with a 99Tc(m) source radionuclide and a non-uniform thorax model. Monte Carlo methods are used to model photon transport and detection. The application of pixel-dependent scatter subtraction factors for scatter compensation is evaluated in an image reconstruction experiment for this simulated myocardial perfusion study. Generalized matrix inverses with noise-dependent regularization are used for image reconstruction. For this simulation, use of a pixel-dependent scatter subtraction factor and a constant scatter subtraction factor are effective for scatter compensation. Activity estimates within the left ventricular myocardium for these two methods are practically the same as those obtained from image reconstructions where the detection of Compton-scattered photons is included in the system matrix. PMID- 15551598 TI - Use of 3D reconstruction to correct for patient motion in SPECT. AB - Patient motion occurring during data acquisition in single photon emission computed tomography (SPET) can cause serious reconstruction artefacts. We have developed a new approach to correct for head motion in brain SPECT. Prior to motion, projections are assigned to conventional projections. When head motion occurs, it is measured by a motion monitoring system, and subsequent projection data are mapped to 'virtual' projections. The appropriate position of each virtual projection is determined by applying the converse of the patient's accumulated motion to the actual camera projection. Conventional and virtual projections, taken together, form a consistent set that can be reconstructed using a three-dimensional (3D) algorithm. The technique has been tested on a range of simulated rotational movements, both within and out of the transaxial plane. For all simulated movements, the motion corrected images exhibited better agreement with a motion free reconstruction than did the uncorrected images. This technique may help to overcome one of the major remaining limitations on image quality and quantitative accuracy in SPECT. PMID- 15551597 TI - A 3D model of non-uniform attenuation and detector response for efficient iterative reconstruction in SPECT. AB - A 3D physical model for iterative reconstruction in SPECT has been developed and applied to experimental data. The model incorporates non-uniform attenuation using reconstructed transmission CT data and distance-dependent detector response based on response function measurements over a range of distances from the detector. The 3D model has been implemented in a computationally efficient manner with practical memory requirements. The features of the model that provide efficiency are described including a new region-dependent reconstruction (RDR) technique. With RDR, filtered backprojection is used to reconstruct areas of the image of minimal clinical importance, and the result is used to supplement the iterative reconstruction of the clinically important areas of the image. The 3D model was incorporated into the maximum likelihood-expectation maximization (ML EM) reconstruction algorithm and tested in three phantom studies--a point source, a uniform cylinder, and an anthropomorphic thorax--and a patient 9Tc(m) sestamibi study. Reconstructed images with the 3D method exhibited excellent noise and resolution characteristics. With the sestamibi data, the RDR technique produced essentially the conventional ML-EM estimate in the cardiac region with substantial time savings. PMID- 15551599 TI - Massively parallel computers for 3D single-photon-emission computed tomography. AB - Since the introduction of the expectation-maximization (EM) algorithm for generating maximum-likelihood (ML) and maximum a posteriori (MAP) estimates in emission tomography, there have been many investigators applying the ML method. However, almost all of the previous work has been restricted to two-dimensional (2D) reconstructions. The major focus and contribution of this paper is to demonstrate a fully three-dimensional (3D) implementation of the MAP method for single-photon-emission computed tomography (SPECT). The 3D reconstruction exhibits an improvement in resolution when compared to the generation of the series of separate 2D slice reconstructions. As has been noted, the iterative EM algorithm for 2D reconstruction is highly computational; the 3D algorithm is far worse. To accommodate the computational complexity, we have extended our previous work in the 2D arena and demonstrate an implementation on the class of massively parallel processors of the 3D algorithm. Using a 16000 processor MasPar machine, the algorithm is demonstrated to execute at 1.24 s/EM iteration for the entire 64 x 64 x 64 cube of 64 planar measurements obtained from the Siemens Orbiter rotating camera operating in the high-resolution mode. PMID- 15551600 TI - In vivo evaluation of a new system for 3D computerized angiography. AB - A new system has been designed and built to validate the concept of 3D computerized angiography (CA). This system can acquire a set of 2D digital subtracted angiography images while rotating around a patient and then, using these images, reconstruct a 3D representation of the opacified vasculature. The design principles and main characteristics of the system are described, with special attention paid to data processing aspects. An initial in vivo evaluation of this system performed on anaesthetized animals and human volunteers is presented. The influence on the quality of the 3D reconstruction of different factors such as volume resolution, estimation method, source trajectory and number of projections is discussed. PMID- 15551601 TI - The synthesis of a new x-ray picture identical in projection to a previous picture using 3D Fourier techniques. AB - A central problem in diagnostic radiology is to compare a new x-ray picture with a previous picture and from this comparison be able to decide whether anatomical changes have occurred in the patient or not. It is of primary interest that these pictures are identical in projection. If not it is difficult to decide with confidence whether differences between the pictures are due to anatomical changes or differences in projection. In this paper we present the geometrical problem and introduce some assumptions that make it possible to find the relative changes in the projection geometry between the exposure of a previous picture and a new picture. The method presented is based on the projection slice theorem (central section theorem). By used 3D Fourier transform techniques we are able to synthesize a new x-ray picture from a set of pictures exposed from a circular orbit above the patient. PMID- 15551602 TI - 3D brain mapping using a deformable neuroanatomy. AB - This paper presents two different mathematical methods that can be used separately or in conjunction to accommodate shape variabilities between normal human neuroanatomies. Both methods use a digitized textbook to represent the complex structure of a typical normal neuroanatomy. Probabilistic transformations on the textbook coordinate system are defined to accommodate shape differences between the textbook and images of other normal neuroanatomies. The transformations are constrained to be consistent with the physical properties of deformable elastic solids in the first method and those of viscous fluids in the second. Results presented in this paper demonstrate how a single deformable textbook can be used to accommodate normal shape variability. PMID- 15551603 TI - 3D reconstruction for a multi-ring PET scanner by single-slice rebinning and axial deconvolution. AB - A three-dimensional (3D) image reconstruction method, which was originally developed for a positron emission tomography (PET) system consisting of two rotating scintillation cameras, has now been implemented for a multi-ring PET scanner with retractable septa. The method is called 'single-slice rebinning with axial deconvolution' (SSAD), and can be described as follows. The projection data are sorted into transaxial 2D sinograms. Correction for the axial blurring is made by deconvolution in the sinograms. To obtain the axial spread functions, which depend on the activity distribution, 2D reconstruction is first made using a limited axial acceptance angle. The final 3D image is obtained by 2D reconstruction of transaxial planes. The method is simple but not approximate, has a modest memory requirement, and can be combined with different 2D techniques. Evaluations by Monte Carlo simulations and phantom studies have been made. PMID- 15551604 TI - Figures of merit for comparing reconstruction algorithms with a volume-imaging PET scanner. AB - For volume-imaging PET scanners, no septa are used to maximize the sensitivity by collecting events oblique to the scanner axis. We answer two questions: (i) how does the performance of an image reconstruction algorithm for a volume-imaging PET scanner depend on its general dimensions? and (ii) at what point is a three dimensional (3D) reconstruction algorithm needed for a volume-imaging scanner, as the axial extent is increased? A 3D reconstruction algorithm will accurately incorporate the oblique events in a reconstruction of the original source distribution. From simulations of an existing volume PET scanner with a maximum axial acceptance angle (+/-alpha) of alpha = 9 degrees, however, we show that the single-slice rebinning algorithm is a good compromise between sensitivity, speed, and accuracy when compared to standard two-dimensional reconstruction (alpha = 1 degrees), and a 3D reconstruction with alpha = 9 degrees. We also show with simulations that a new scanner with alpha = 27 degrees requires 3D reconstruction in order to achieve maximum sensitivity without unacceptable losses in accuracy. Measurements of scanner performance are based on a series of figures of merit that characterize image quality and quantitative accuracy measured from a set of simulated test phantoms. PMID- 15551605 TI - Implementation of an accelerated iterative algorithm for cone-beam SPECT. AB - In this paper we describe the implementation of an accelerated iterative reconstruction algorithm (AIRA) for cone-beam (CB) projections using a single circular orbit in single-photon-emission computed tomography (SPECT). This algorithm is a modified maximum-likelihood-expectation-maximization (ML-EM) algorithm and several approaches have been used to accelerate the reconstruction process. These approaches include: (i) the use of ordered subsets; (ii) the use of active areas and volumes; and (iii) the storing in memory of the transition vector for a given ray (during the forward projection step). This algorithm, which compensates for collimator geometric sensitivity variation as a function of position and makes uniform attenuation corrections has been evaluated using experimentally acquired phantom data. The results demonstrate a two-orders-of magnitude decrease of the computational time of this algorithm over the conventional ML-EM algorithm with similar convergence properties. PMID- 15551606 TI - Apical limit of root canal instrumentation and obturation, part 1. Literature review. AB - One of the major controversies in root canal therapy concerns the apical limit of instrumentation and obturation. The results of longitudinal prognostic studies, basic anatomical knowledge of the apical third of the root canal, and the histological pulp reaction to caries progression demonstrated the presence of a vital apical pulp remnant, even in the presence of a periapical lesion. Finally necrosis and bacteria establish themselves in the periapical lesion. All valid prognosis studies confirm the practice of staying short of the apex with a homogeneous obturation to obtain the highest success rate of 90-94% (when done by or under supervision of specialists; results in the general population had a failure rate greater than 50%). The location of the apical foramen(ina) related to root canal treatment most frequently ends short of the apex, often by several millimetres. PMID- 15551607 TI - Apical limit of root canal instrumentation and obturation, part 2. A histological study. AB - The results of an in vivo histological study involving apical and periapical tissues following root canal therapy after different observation periods demonstrated the most favourable histological conditions when the instrumentation and obturation remained at or short of the apical constriction. This was the case in the presence of vital or necrotic pulps, also when bacteria had penetrated the foramen and were present in the periapical tissues. When the sealer and/or the gutta-percha was extruded into the periapical tissue, the lateral canals and the apical ramifications, there was always a severe inflammatory reaction including a foreign body reaction despite a clinical absence of pain. PMID- 15551609 TI - A histopathological study of the morphological changes at the apical seat and in the periapical region after irradiation with a pulsed Nd:YAG laser. AB - In this study, the effects of pulsed Nd:YAG laser irradiation on the apical seat and periapical region of instrumented root canals were investigated. One hundred and ninety-two teeth, including incisors and molars (239 root canals), in 18 healthy adult beagle dogs were used. After extirpation of the pulp under anaesthesia, each root canal was shaped up to at least a size 40 K-file, and then laser irradiated using the following parameters: 1 W, 15 pps for 2, 3, and 4 s; 2 W, 15 pps for 2 s. The effectiveness of debris removal and carbonization on the root canal walls immediately laser irradiation were examined and inflammation of the periapical region at 1, 2, and 4 weeks after laser irradiation was examined using light microscopy. Effective debris removal was observed in most cases in the laser-treated groups and was dependent on irradiated energy. Carbonization was observed in the irradiated root canals except at the parameter of 1 W for 2 s. Periapical inflammation was observed at 1 W for 2 s to the same extent as in the control group (P > 0.01). These results suggest that the pulsed Nd:YAG laser is useful for debris removal with no adverse effects immediately after extirpation of the pulp if appropriate parameters are selected. PMID- 15551608 TI - Two canals in mesiobuccal roots of maxillary molars. AB - The purpose of this study was to examine extracted root canal treated maxillary molars cleared for: (i) the presence of a mesiopalatal (MP) canal in both first and second molars, (ii) the extension of MP canal from the pulp to the apical area, and (iii) the incidence of two foramina in the MB root. Seventy-two extracted maxillary molars (42 first and 30 second molars) were root canal treated by graduate students. The mesiobuccal (MB) roots of these samples were rendered transparent using a clearing technique. The results demonstrated that 52.3% of first and 40% of second molars had two canals obturated in the MB root. After clearing the same roots, the presence of MP canals rose to 80.9% and 66.6%, respectively. The MP canals were root canal treated as far as the foramen in 35.2% of first and 35% of second molars. However, after making them transparent, 91.1% and 90% showed the presence of this canal to the anatomical apex. The MB roots of the root canal treated first molars showed the presence of two foramina in 47% of cases but in 88.2% after clearing. The second molars showed 50% and 70%, respectively. The differences between root canal treated teeth before and after clearing were significantly different in almost all comparisons in both first and second molars. The only exception was when the presence of two foramina was compared before and after the clearing process in second molars (McNemar test P < 0.05). PMID- 15551611 TI - Linking reimbursement and quality. PMID- 15551612 TI - The economic impact of childhood obesity. PMID- 15551613 TI - An evolving strategy to increase the number of minority and low-income patients referred for transplantation. PMID- 15551614 TI - Yes I can! PMID- 15551615 TI - Vancomycin intermediate and resistant Staphylococcus aureus. What the nephrologist needs to know. AB - Individuals undergoing hemodialysis may be at increased risk for emerging antimicrobial resistance from vancomycin-intermediate Staphylococcus aureus (VISA) and vancomycin-resistant S. aureus (VRSA). The laboratory detection of VISA and VRSA is challenging and requires the use of well-thought-out algorithms. Newly available antimicrobials such as quinipristin/dalfopristin, linezolid, and daptomycin, as well as older drugs such as trimethoprim-sulfamethoxazole appear to be active against recent strains of VISA and VRSA. Prevention of VISA and VRSA necessitates determining the appropriateness of vancomycin use in renal patients and giving priority to infection control precautions in both inpatient and outpatient settings. Because most VISA and all VRSA to date have arisen from endemic methicillin-resistant S. aureus (MRSA), and in the case of VRSA have acquired genes from vancomycin-resistant enterococci (VRE), the emergence of VISA and VRSA should provide renewed motivation for the containment of MRSA and VRE transmission in the hemodialysis population. PMID- 15551616 TI - Structure and content of chronic kidney disease information on the World Wide Web: barriers to public understanding of a pandemic. AB - Chronic kidney disease (CKD) is a pandemic and the need to inform those at risk has never been more important. The World Wide Web (WWW) is no w considered a key source of health information, but the quality and utility of this information has been challenged. In this article, we assess structural, content, and linguistic barriers to accessed CKD information and discuss the implications of limited Internet access to communicating health. Technical (number of hyperlinks), content (number of six core CKD and risk factor information domains included), and linguistic (readability and variation in readability) barriers were assessed for websites offered by 12 kidney disease associations. The Flesch Reading Ease Index method was used to estimate readability scores, and variation in the readability of information was assessed. Eleven websites met inclusion criteria. Six of 11 websites provided information in all 6 domains of CKD information. A mean of 4 hyperlinks (range 3-5) was clicked before CKD information was available and a mean of 6 hyperlinks (range 4-12) was clicked to access all available CKD information. Mean readability scores for all six domains of CKD information exceeded national average literacy skills and far exceeded the 5th grade level readability desired for informing vulnerable populations. Information about CKD and diabetes consistently had higher readability scores. The WWW currently has little utility for informing populations at greatest risk for CKD. Barriers to accessing CKD information on the WWW are socioeconomic, technical, and linguistic. Having lower socioeconomic status, less access to computers and the WWW, multiple website hyperlinks, incomplete information, difficult readability, and significant variation in readability of CKD information on the WWW are social, structural, and content barriers to communicating CKD information. This may contribute to the growing epidemics of diminished public understanding about CKD, and disparities in CKD health status experienced by racial/ethnic minority populations globally. PMID- 15551617 TI - KI study: older patients with CKD are costly as they transition to dialysis. PMID- 15551618 TI - Reflections on a tragedy in Canada. PMID- 15551619 TI - Fistula first: an update for renal providers. PMID- 15551620 TI - FMCNA says data shows higher risk of mortality among patients on reuse. Industry questions bias in reporting. PMID- 15551621 TI - Reuse: what does the industry think? PMID- 15551623 TI - The quality imperative: why the kidney community must take charge. PMID- 15551624 TI - [Comparative analysis of commercial renal diets for cats, with emphasis on their fatty acid composition]. AB - The objective of this study was to compare renal diets for cats that are commercially available in The Netherlands. The diets were analysed and their composition compared with a guideline proposed by the authors. Special attention was paid to the fatty acid composition of the diets and the role of fatty acids in the progression of chronic renal failure. On the basis of a total score for nutrient levels relative to the guideline recommendations, the diets were ranked as to expected efficacy. PMID- 15551625 TI - ['Sock suffering']. PMID- 15551626 TI - [VWA looks after the law of animal experimentation (WOD)]. PMID- 15551627 TI - [Permanent representative for the EU Ton Akkerman finds pre-judgement about admitting new member states to be nonsense. Text and photos by Susan Umans]. PMID- 15551628 TI - [Frontline contra-indicated in the rabbit]. PMID- 15551629 TI - [Acute zinc poisoning in a dog]. AB - Zinc-induced haemolytic anaemia was diagnosed in a young dog. The origin of the zinc intoxication was the ingestion of a toy material which contained a high percentage of zinc. The level of zinc in the liver and kidneys was resp. 1050 and 1320 ppm. PMID- 15551630 TI - [Goodwill: adjustment of the current method]. PMID- 15551631 TI - [Work disability prevention: report of a pilot project]. PMID- 15551632 TI - [Self regulation in veterinary medicine]. PMID- 15551633 TI - [VI--Decubitus ulcer physiopathology]. PMID- 15551634 TI - [9--Clinical aspects: Decubitus ulcers. Local treatment of decubitus ulcers]. PMID- 15551635 TI - [Better nursing qualifications, benefits for patients]. PMID- 15551636 TI - [Care of the diabetic foot. The contribution of each nurse...]. PMID- 15551637 TI - [Care of the diabetic foot. Some recommendations on terms of management? Interview by Bernadette Fabregas]. PMID- 15551638 TI - [Care of the diabetic foot. A well-established therapeutic strategy]. PMID- 15551639 TI - [Care of the diabetic foot. The concept of the "wound healing center"]. PMID- 15551640 TI - [The nurse/nurse's aide dyad, what is the reality?]. PMID- 15551641 TI - [Nurse's aides and nurses, what role in the European scale?]. PMID- 15551642 TI - [The nurse/nurse's aide relationship, collaboration yes, delegation no]. PMID- 15551643 TI - [February 11 2002 decree concerning nursing competences]. PMID- 15551644 TI - [The nurse/nurse's aide dyad, what is the reality? Which collaboration in drug therapy distribution?]. PMID- 15551645 TI - [The nurse/nurse's aide dyad, what is the reality? One or the other, what power for what role?]. PMID- 15551646 TI - [The nurse/nurse's aide dyad in hemodialysis]. PMID- 15551647 TI - [The nurse/nurse's aide dyad, what is the reality? A natural complementary relationship]. PMID- 15551648 TI - [The nurse/nurse's aide dyad, what is the reality? For better...and for worse!]. PMID- 15551649 TI - [The nurse/nurse's aide dyad, what is the reality? Bibliography]. PMID- 15551650 TI - Issues in developing the medical scientist, part 2: fostering research among medical students (interview with Michael John McPhaul, MD). PMID- 15551651 TI - Consequences of cold ischemic injury of kidneys in clinical transplantation. AB - Cold storage allows the effective utilization of organs retrieved for transplantation. However, prolonged cold ischemia contributes to organ damage and increases patients' morbidity and mortality and health care cost. Using recent data from the United Network for Organ Sharing, this paper examines the outcomes of live donor and deceased donor kidney transplants in relation to cold ischemia time. PMID- 15551652 TI - New insights into the cellular and molecular mechanisms of cold storage injury. AB - Solid organ grafts, but also other biologic materials requiring storage for a few hours to a few days, are usually stored under hypothermic conditions. To decrease graft injury during cold storage, organ preservation solutions were developed many years ago. However, since then, modern biochemical and cell biologic methods have allowed further insights into the molecular and cellular mechanisms of cold storage injury, including further insights into alterations of the cellular ion homeostasis, the occurrence of a mitochondrial permeability transition, and the occurrence of free-radical-mediated hypothermic injury and cold-induced apoptosis. These new aspects of cold storage injury, which are not covered by preservation solutions in current clinical use and offer the potential for improvement of organ and tissue preservation, are presented here. PMID- 15551653 TI - Immune responses in kidney preservation and reperfusion injury. AB - Organ preservation and reperfusion injury have significant detrimental effects on both short- and long-term organ function. Ischemia reperfusion injury (IRI) underlies organ transplant dysfunction, myocardial infarction, stroke, and shock. Multiple molecular pathways are engaged in reactive oxygen production, apoptosis, signaling, and tissue regeneration. There has been an increased understanding of the important role of immune and inflammatory pathways in IRI, both in humans and in experimental models. Both cellular and soluble components of the immune system are directly activated during IRI, and there is evidence that immune mediators directly contribute to organ dysfunction. Immune activation during IRI likely underlies the enhanced immunogenicity of ischemic organs, with resultant increased rejection and fibrosis. Novel human therapies targeting T and B cells for classic immune diseases can now be considered to prevent and treat IRI. Organ preservation injury and cold ischemia could well have distinct pathophysiology from warm IRI and represent an opportunity to develop improved preservation methods. PMID- 15551654 TI - Cold ischemic organ preservation: lessons from natural systems. AB - Mammalian hibernators offer natural models for investigating solutions to the metabolic injuries that accrue during cold ischemic storage of human organs removed for transplant. Knowledge of the biochemical mechanisms that regulate and stabilize metabolism to ensure long-term viability in the hypometabolic, hypothermic state of hibernation could lead to applied treatments that could increase the time that excised organs can be maintained in cold storage and/or improve recovery of function after implantation. New research has documented the widespread role of reversible protein phosphorylation control of metabolism in achieving the coordinated suppression of metabolic rate that greatly extends viability during torpor. Analysis of hibernation-induced gene expression is proving to be of crucial importance for identifying the genes and proteins that are up-regulated to address organ-specific concerns during torpor. In particular, the power of complementary deoxyribonucleic acid (cDNA) array screening is identifying families of proteins that are up-regulated during hibernation (eg, serpins, heat shock proteins, antioxidants, membrane transporters) and highlighting previously unrecognized areas of cellular metabolism as contributing to the hibernation phenotype. These offer new targets for innovative applied treatments that could enhance cytoprotection and cold ischemia survival of organ explants. PMID- 15551655 TI - Preconditioning strategies to limit graft immunogenicity and cold ischemic organ injury. AB - During the transplant process, the graft is exposed to numerous events, which may enhance its immunogenicity. In particular, factors related to brain death, such as hemodynamic instability and systemic release of cytokines, cold preservation on harvesting, and reperfusion injury, are known to accumulate in harm, conveying a proinflammatory state to the graft before transplant. Alloimmune reactivity is initiated when the host immune system detects non-self-antigens in the context of "danger signals." Eliminating these danger signals by modifying the graft before transplant has the potential to attenuate the alloimmune response. The molecules, which mediate danger signals, have not yet been fully identified. Free oxygen radicals and interferon-gamma are important candidates. One of the most important protective mechanisms against oxidative stress is the heme oxygenase 1 system. Up regulation of heme oxygenase 1 in grafts has been shown to prevent ischemia reperfusion damage and improve long-term graft survival in various transplant models. The benefit of blocking the action of interferon-gamma in kidney transplants is less clear because the compound plays such a complex and pivotal role in the immune response, and experimental data with interferon-gamma receptor knockout mice are conflicting. It has recently become clear that catecholamines are important graft-modifying agents. Dopamine is capable of stimulating the induction of protective enzymes like heme oxygenase-1 (HO-1) rendering the organ more resistant to the insult of ischemia/reperfusion and inflammation. Retrospective clinical data suggest that treatment of brain-dead organ donors with catecholamines is associated with less rejection and a better long-term graft survival of kidneys transplanted from these donors. Catecholamines can also modulate cytokine production and prevent cold-induced damage. Other substances, such as proteoglycans and phosphatidylethanolamine-bound hyaluronic acid, may interfere with the actions of interferon-gamma. Further studies of these compounds in experimental animal models and in prospective randomized clinical trials will help establish their efficacy in donor pretreatment. It is important to underscore that donor pretreatment will have great advantages for the recipient because an improved long-term graft survival could thus be achieved cost-efficiently and without great effort or side effects. PMID- 15551656 TI - Effect of adventitial dissection in brachiobasilic arteriovenous fistulae opened in children as vascular access for hemodialysis on patency and maturation of fistulae. AB - BACKGROUND: Recently, therapy of pediatric patients with chronic renal failure has been carried out using hemodialysis, peritoneal dialysis, or renal transplant. In this study, we prospectively investigated the role of adventitial dissection (performed on brachiobasilic arteriovenous fistulae in the antecubital area) in the patency rate, maturation, and early initiation of dialysis among pediatric cases undergoing chronic hemodialysis. METHODS: Thirty children (7 male and 23 females) were included in this study. They were operated on at the Department of Cardiovascular Surgery of Alsancak State Hospital and Dokuz Eylul University between March 2001 and June 2003. Their mean age was 12.3 +/- 2.9 years (range 7-17 years), and their mean weight was 25.5 +/- 7.9 kg (range 16-44 kg). Children who underwent only brachiobasilic arteriovenous fistula operation were included. Group 1 (n = 15) consisted of those who underwent adventitial dissection, and group 2 (n = 15) consisted of those not receiving the adventitial dissection procedure. RESULTS: The procedure was conducted by the same surgeon, and follow-up examinations were done at the Department of Pediatric Nephrology by investigators masked to treatment groups. CONCLUSION: There were no significant differences in age, sex, existence of preoperative hypertension, diameter of vein measured preoperatively, and first day of dialysis between the groups. In 29 cases, a palpable thrill was noted at the operation site. Hemodialysis had been initiated after the fourth week, when the fistulae had matured. The mean duration of follow-up was 12.53 +/- 8.98 months in group 1 and 11.85 +/- 7.55 months in group 2 (p = .880). In group 1, one case developed fistula infection in the second month and one case developed lymphatic drainage. In group 2, one case developed early thrombosis, one case developed hematoma, and one case developed transient ischemia owing to arterial spasm in the ipsilateral limb. One case in each group developed minimal edema in the forearm, which was treated conservatively. The primary patency rate was 93.3% in both groups, whereas the secondary patency rate was 100% in group 1. Adventitial dissection performed on the vein during arteriovenous fistula formation does not result in any additional benefit with respect to fistula maturation, early initiation of dialysis, and patency. Among pediatric patients with chronic renal failure, in cases of inappropriate forearm veins, brachial arteriovenous fistulae performed with a meticulous surgical technique should be the procedure of choice because it is associated with minimal complications and a high patency rate. PMID- 15551658 TI - Nursing's ethos of caring and its support for a single-tiered health-care system. PMID- 15551657 TI - Simvastatin induces death of multiple myeloma cell lines. AB - BACKGROUND: Accumulating reports indicate that statins widely prescribed for hypercholesteromia have antineoplastic activity. We hypothesized that because statins inhibit farnesylation of Ras that is often mutated in multiple myeloma (MM), as well as the production of interleukin (IL)-6, a key cytokine in MM, they may have antiproliferative and/or proapoptotic effects in this malignancy. METHODS: U266, RPMI 8226, and ARH77 were treated with simvastatin (0-30 microM) for 5 days. The following aspects were evaluated: viability (IC50), cell cycle, cell death, cytoplasmic calcium ion levels, supernatant IL-6 levels, and tyrosine kinase activity. RESULTS: Exposure of all cell lines to simvastatin resulted in reduced viability with IC50s of 4.5 microM for ARH77, 8 microM for RPMI 8226, and 13 microM for U266. The decreased viability is attributed to cell-cycle arrest (U266, G1; RPMI 8226, G2M) and cell death. ARH77 underwent apoptosis, whereas U266 and RPMI 8226 displayed a more necrotic form of death. Cytoplasmic calcium levels decreased significantly in all treated cell lines. IL-6 secretion from U266 cells was abrogated on treatment with simvastatin, whereas total tyrosine phosphorylation was unaffected. CONCLUSIONS: Simvastatin displays significant antimyeloma activity in vitro. Further research is warranted for elucidation of the modulated molecular pathways and clinical relevance. PMID- 15551659 TI - Rethinking the research-practice gap: relevance of the RCT to practice. PMID- 15551660 TI - Evaluating the effectiveness of interventions: exploration of two statistical methods. AB - Repeated measures designs are often used to evaluate the effectiveness of interventions. In these designs, the outcomes are measured on several occasions before and after implementation of the intervention. Two statistical methods, the repeated measures analysis of variance (RM-ANOVA) and hierarchical linear models (HLM), can be used to analyze the data. The authors provide an overview of the statistical models underlying RM-ANOVA and HLM and discuss the strengths and limitations of each. They propose that the 2 methods are complementary in determining the effectiveness of interventions. PMID- 15551661 TI - Intersectionalities of influence: researching the health of immigrant and refugee women. AB - There is a growing recognition of the complexity surrounding multiple axes or dimensions of social identity and how they intersect to influence the health of immigrant and refugee women. The concept of intersectionalities of influence is particularly relevant in addressing diversity in nursing research. The purpose of this paper is to theorize and operationalize the concept in mental health promotion research with immigrant and refugee women. At the conceptual level, the authors propose an approach to inquiry that is informed by critical scholarship and draws from postcolonial and feminist perspectives. At the operational level, they apply an ecosystemic framework to help locate individual health issues within the familial, community, and social realms. They introduce Participatory Action Research as a way of putting these concepts into action within the research process. Their aim is to introduce a new way of inquiry that can benefit immigrant and refugee women while furthering the nursing agenda for community based research. PMID- 15551662 TI - Fall-prevention programs for the elderly: a Bayesian secondary meta-analysis. AB - A secondary meta-analysis of programs to reduce falls in the elderly is undertaken to demonstrate a Bayesian analysis. The Bayesian statistical tradition is carefully distinguished from the standard Neyman-Pearson-Wald (NPW) statistical tradition. In the 12 studies, the logit effect size is used to compare treatment groups using a prevention program to control groups without a program. To contrast the Bayesian analysis, independent-effects and fixed-effect meta-analyses are first conducted in the NPW tradition. This is followed by Bayesian independent-effects and fixed-effect meta-analyses that numerically replicate the NPW results but have conceptually different interpretations. The final analyses comprise Bayesian random-effects and predictive meta-analyses. These results differ numerically from all the previous meta-analyses and conceptually from the NPW meta-analyses. The random-effects analysis allows for heterogeneity in the effect sizes. The predictive analysis yields the distribution of a new, out-of-sample effect size, which accommodates not only the heterogeneity of the effects but also the imprecision in the parameter estimates. This last analysis shows that the effectiveness of new fall-prevention programs is less definitive than that found in the sample. Bayesian statistical methods are particularly well-suited for the complexities of nursing science studies. PMID- 15551663 TI - The scale product technique as a means of enhancing the measurement of patient satisfaction. AB - Measurement of patient satisfaction has long been hampered by two resolvable problems: a lack of content validity in commonly used instruments, and a lack of variability in satisfaction scores when these same instruments are used. Most patient satisfaction instruments have been developed from the perspective of the provider or institution rather than that of the patient, creating a situation of questionable content validity for these measures. Additionally, most patient satisfaction measures yield data that are invariant and consistently positively biased. Both of these problems can be addressed methodologically--through tool development using a qualitative method designed to obtain the patient's perspective, and through the use of the scale product technique to decrease the effect of acquiescence, thereby increasing variability in item responses. PMID- 15551664 TI - Are we missing anything? Pursuing research on attrition. AB - Attrition, or loss of participants over the course of a study, presents a significant threat to the integrity of a longitudinal research study and theory development resulting from the study. Although there has been a recent resurgence of interest in attrition, it is an underreported and understudied phenomenon despite its potential to introduce bias. Internal and external validity, reliability, and statistical validity are all impacted by a small sample and, most likely, a non-randomness in the study sample. Development of a theory of attrition will assist researchers in development of sampling strategies that will enhance the quality of their data in longitudinal designs. An ecological theoretical model of research participation is proposed and described. PMID- 15551665 TI - An exploration of participants' treatment preferences in a partial RCT. AB - The partial RCT is a modified design meant to account for participants' treatment preferences. Little is known about the factors that influence such preferences. This study investigated the extent to which participants are willing to be randomized and the factors that affect their treatment preferences. The quantitative and qualitative data collected as part of a partial RCT evaluating 2 behavioural interventions for insomnia were analyzed. Of the participants, 75% were unwilling to be randomly allocated to a treatment group. This figure increased to 90% after information on the 2 interventions was provided. Participants' knowledge of the treatment, its suitability, and their perceived ability to adhere to it influenced their treatment preference. A few participants who had no strong preference and who required treatment were willing to be randomized. The findings raise questions about the impact of treatment preference and allocation procedures on the validity of study conclusions. PMID- 15551666 TI - Advancing patient-centred care through knowledge development. AB - The call for health-care services that are patient-centred raises the need for knowledge development in both the conceptual and empirical domains. The definitions and operational elements of patient-centred care present a variety of conceptual issues. A common element in all definitions is accommodation of patient wants, preferences, and expectations. In the research domain, intervention studies face both design and measurement challenges. These include the development of interventions that are patient-centred or tailored for both patient characteristics and the environment in which they will be delivered. By addressing these critical issues, nursing can play a key role in advancing intervention science and knowledge development in the domain of patient-centred care. PMID- 15551667 TI - Personal Construct Theory: a strategy for the study of multidimensional phenomena in nursing. PMID- 15551668 TI - The mental health of moving asylum seekers from 'temporary' to 'permanent' protection visas: it's much more than a quick political fix. PMID- 15551669 TI - Cultural diversity: a challenge for community nurses. AB - Cultural diversity is a challenge for community nurses and can present many difficulties in the provision of quality nursing care and in achieving optimal health outcomes. In this paper seven enabling principles are identified and their application to diverse groups discussed. These include: willingness to recognize expert family skills and knowledge, acknowledging own and other nurses' strengths and weakness, taking time to establish rapport and acceptance, assessing influences on health and health care, providing care that is culturally appropriate, developing cultural competent practice and advocating for CLD appropriate resources and expertise. The aim of this paper is to assist community nurses to provide nursing care congruent with the many lifeway practices encountered in the community. Diverse cultural groups with distinguishing dominant characteristics of ethnic orientation, aboriginality, and disability are selected as examples to demonstrate the application of the principles. PMID- 15551670 TI - Being active supports client control over health care. AB - The purpose of this study was to identify how healthcare clients achieve and maintain a sense of control over their health. The literature review conducted refers to: (i) key definitions of control, (ii) locus of control, and (iii) control and wellbeing. Participants with a range of acute and chronic health conditions and who had been hospitalised at some point were selected for the study. Symbolic interactionism (Blumer, 1969) and modified grounded theory of Strauss & Corbin (1998) provided the frameworks for this study. During the six month study period, data were collected from sixty participants and included interviews, participant observation, reviewing participants' records (nursing care plans, nursing notes and case histories), the nursing units' philosophy, organisational charts, policies and procedures, annual reports, consumer brochures and any other relevant information sources. Findings from the study indicated that participants moved from feeling vulnerable to having a sense of control through to being purposefully active. Vulnerability was associated with: (i) having limited choices in respect to their health, (ii) lacking adequate health information to make choices, (iii) being ignored by health providers with respect to their needs, and (iv) lacking friend/family supports. Purposefully activating was associated with three major categories: (i) reflecting, (ii) being self-determiningly involved and (iii) normalising. Findings from this study could be used by health care clients who want a sense of control over their health care, and also by health care providers who wish to support clients in the healthcare process. PMID- 15551671 TI - A case for consideration of cultural diversity in heart failure management--Part 1: Rationale for the DISCOVER Study. AB - Heart failure is a condition increasing in prevalence and responsible for high health care utilization, morbidity and mortality. Randomised controlled trials of nurse-coordinated interventions have determined self-care and the incorporation of the patient and their family in care planning as critical elements of service delivery. Coping with a chronic illness, such as heart failure, forces the individual to adjust to changed physical, social and emotional functioning and to modify their lifestyle accordingly. Clinicians increasingly use models of care that focus care delivery on the community setting. In order to develop strategies to assist patients and their families with self-care it is important that clinicians understand the health-care seeking behaviours of all individuals targeted in the community. Australia is a culturally diverse nation, yet evaluations of models of care have been undertaken largely in individuals from predominately Anglo-Celtic origins. The end result of this approach is failure to understand the full range of diverse perspectives that individuals hold that can have an impact on self-care behaviours. Consideration of cultural diversity should extend beyond language to a broader appreciation of cultural values, health seeking beliefs and engagement of culturally unique communities. The 'Understanding the cultural experiences of individuals with chronic heart failure (CHF) in South East Health (DISCOVER) Study' seeks to uncover information on the health patterns, information needs and the adjustment process for overseas-born individuals with heart failure. Such information will assist clinicians to tailor health care service delivery and ensure the delivery of appropriate, quality care. This manuscript provides the background, rationale and methods for this study. PMID- 15551672 TI - Targeting community-dwelling urinary incontinence sufferers: a multi-disciplinary community based model for conservative continence services. AB - This paper presents an argument that there is a need to provide services that target community-dwelling incontinence sufferers, and presents a demonstration case study of a multi-disciplinary, community-based conservative model of service delivery: The Waterworx Model. Rationale for approaches taken, implementation of the model, evaluation and lessons learned are discussed. In this paper community dwelling sufferers of urinary incontinence are identified as an underserved group, and useful information is provided for those wishing to establish services for them. The Waterworx Model of continence service delivery incorporates three interrelated approaches. Firstly, client access is achieved by using community based services via clinic and home visits, creating referral pathways and active promotion of services. Secondly, multi-disciplinary client care is provided by targeting a specific client group, multi-disciplinary assessment, promoting client self-management and developing client knowledge and health literacy. Finally, interdisciplinary collaboration and linkages is facilitated by developing multidisciplinary assessment tools, using interdisciplinary referrals, staff development, multi-disciplinary management and providing professional education. Implementation of the model achieved greater client access, improvement in urinary incontinence and client satisfaction. Our experiences suggest that those suffering urinary incontinence and living in the community are an underserved group and that continence services should be community focussed, multi-disciplinary, generalist in nature. PMID- 15551673 TI - Exploring the complexities of prostate cancer screening with a view to supporting informed consent. AB - Men request to be screened for prostate cancer because they believe they are exhibiting responsible health promotion behaviour and there are definite benefits from the early detection of the disease. This belief about the benefits is in contrast to several national guidelines recommending that screening for prostate cancer not be done. Despite the guidelines men continue to request to be screened and doctors continue to supply screening tests to asymptomatic males. The lack of an appropriate screening test has been a key factor in supporting recommendations not to screen. However, recent studies show improved accuracy in the use of serum prostate specific antigens (PSA) as a screening tool. This implies that a revision of the guidelines might soon be appropriate. It is important that nurses and other health professionals are kept abreast of developments in prostate screening to assist men with their screening options to ensure their fully informed consent. PMID- 15551674 TI - Mental health knowledge in residential aged care: a descriptive review of the literature. AB - Health care staff knowledge of mental health is vital in the provision of quality care for older people in residential aged care settings. This paper aims to describe mental health knowledge competence of health care staff in residential aged care through a review of existing literature and to explore the link between knowledge, attitudes and education about older people and mental health. A literature review was conducted using electronic databases and library catalogues to identify articles published in English during the period 1982-2002. There is insufficient evidence within the literature to draw conclusions about staff knowledge levels in relation to mental health, however, the literature identified a link between continuing education, knowledge levels and staff attitudes to older people with mental health disorders. Future studies are needed to investigate existing levels of mental health knowledge among health care staff in residential aged care and to identify and evaluate strategies to enhance their ability to provide care for this population. PMID- 15551675 TI - Exploring bereavement care in inpatient settings. AB - This paper compares the attitudes and practices of health professionals to the provision of family-centred bereavement care in different Australian inpatient settings: palliative care services, acute care and long stay residential aged care settings. Semi-structured interviews (88) were conducted with doctors, nurses, pastoral carers and allied health staff. Comprehensive bereavement care was not being provided or resourced in the acute and aged care hospitals to the same level as that provided in palliative care services. Residential care provided continuity of care and good support for grieving relatives and other residents. A structured bereavement program is needed in all inpatient settings with palliative care patients, along with resource and education support for health professionals. PMID- 15551676 TI - Management responsibilities in the retirement village industry: a New Zealand study. AB - The Retirement Village/Rest Home industry is growing rapidly. Many of those fulfilling managerial roles within this industry do not appear to have management qualifications or prior managerial experience outside the public sector health system. If the industry is to effectively manage anticipated growth, managers may need to update their qualifications and knowledge base to ensure that they possess the skills and abilities required to meet changing business needs. This paper reports on an exploratory study designed to gather information that identifies the primary duties and responsibilites of retirement village managers and nurse managers, and their essential and desirable skills, attributes, and characteristics. PMID- 15551677 TI - Flushing an i.v. line: a simple but potentially costly procedure for both patient and health unit. AB - This observational study investigated the intravenous (i.v.) medication administration technique of Registered Nurses (n = 47) using the injection port of a concurrent intravenous infusion. Study findings showed that the majority of nurses adhered to the most common although debatable recommended guideline in stopping the concurrent infusion and slowly administering the medication in the syringe over 3-5 minutes, prior to flushing the line quickly. Nurses failed to account for the dead space in the i.v. line when calculating the administration rate for pre and post medication flush. Similarly, the benefits of continuing the concurrent infusion when administering intravenous medication were not taken into consideration. Based upon these study findings, we recommend changes to procedural guidelines for i.v. medication. PMID- 15551679 TI - Nurse-academics' scholarly productivity: perceived frames and facilitators. AB - The reward system within Universities remains focused on research, with a benchmark of scholarly productivity, especially in relation to promotion. Despite their relative newness to the tertiary system, nurse academics are judged by the same standards as other disciplines. This study sought to examine factors that constrained and/or facilitated scholarly productivity. The study used a questionnaire survey technique to establish current productivity levels, and frame and facilitating factor theory and analysis to identify major constraints and facilitators. Findings from the study were that the unremitting nature of teaching, course coordination and university service workloads interact to the detriment of research and writing. Facilitating factors included a departmental culture that values and supports research, in conjunction with tangible support from University management. Mentorship was viewed as desirable, but often not available. An environment that more actively prioritises, fosters and supports academic scholarly productivity is needed. PMID- 15551678 TI - Aneurysmal subarachnoid haemorrhage--Part 2: Post-operative nursing management diagnosis, complications and treatment: a composite case study. AB - Patients who have been diagnosed with an Aneurysmal Subarachnoid Haemorrhage (ASAH) frequently require surgical intervention to prevent further complications. Due to the complex nature of ASAH it is imperative to be meticulous in monitoring the patient's neurological condition. This is even more relevant post operatively as complications such as cerebral vasospasm peak incidence is between days four to fourteen and as well as complications of surgical and medical intervention may become evident. PMID- 15551681 TI - Diagnostic images flowing among clinicians. PMID- 15551680 TI - Factors affecting the job stress and job satisfaction of Australian nurses: implications for recruitment and retention. AB - Against a background of nurse shortages in Australian hospitals, a significant challenge facing the healthcare sector is the recruitment and retention of nurses. The job stress and job satisfaction of nurses have been associated with recruitment and retention. The aim of this study is to consider two factors that may contribute to the job satisfaction and job stress of nurses: social support and empowerment. Using a sample of 157 registered nurses in a private hospital in Melbourne, Australia, we found that social support derived from the nurse's supervisor and work colleagues lowered job stress and at the same time increased job satisfaction. The presence of nurse empowerment, meaning, impact, competence and self-determination, also lowered job stress and increased job satisfaction. Finally, we discuss contributions of this study and implications for recruitment and retention of nurses in the health sector. PMID- 15551682 TI - Data mining: solving care, cost capers. PMID- 15551683 TI - Making the Internet safe for health data. PMID- 15551684 TI - Doctors to PDAs: 'Check, please'. PMID- 15551685 TI - Readers perspectives... Biometrics technology. PMID- 15551686 TI - On ethical peer review and publication: the importance of professional conduct and communication. AB - Editors and reviewers provide an invaluable service to the nursing profession and are key to the promotion of nursing scholarship, open inquiry uncensored by opinion or bias, and further development of nursing's body of knowledge. By being accessible and promoting an effective, constructive review process, editors serve not only the nurses who write for them, but also their respective journals and readership. Authors deserve and expect professional and ethical behavior from peer reviewers and those in leadership who make final judgments about the publication status of their manuscripts. PMID- 15551687 TI - Direct links to author guidelines. PMID- 15551688 TI - [The international debate on the effectiveness of health promotion: where does it come from? Why is it so important?]. AB - This paper aims at positioning the issue of health promotion effectiveness in the context of the international debate around the utilisation of evidence-based practices. In a sociological and historical perspective, it analyses how the evidence-based practices approach has confronted the field of health promotion, especially since the second half of the 1990's. It argues that this has occurred in the context of the evaluative concerns put forward by governments all over the world, in order to downsize their publicly funded health systems. After providing definitions and some historical context, five methodological and political issues raised by the idea of utilizing evidence-based practices in health promotion are presented. The paper concludes that this approach is far from a panacea but has potential limited utilization for certain kinds of health promotion practices. Alternative suggestions on how to establish "best practices" in this field are also offered. PMID- 15551689 TI - [Using evidence to assess the effectiveness of health promotion programs: a few fundamental issues]. AB - This paper first discusses more generally the notion of "evidence" and then applies it to the field of health promotion. It is argued that health promotion has not yet reached the status of a fully recognised and recognisable discipline, borrowing many of its theoretical and methodological approaches from apparatuses to a wide variety of established other disciplines. Consequently, developing "evidence-based" practices in health promotion, as in biomedicine for example, is problematic on many counts and leaves three major issues unsolved. These issues are addressed and the authors conclude on an optimistic note, despite the serious problems that still remain to be solved. PMID- 15551690 TI - [Towards a framework to assess the effectiveness of community-based health promotion initiatives: recent North-American developments]. AB - The current systematic reviews to assess the effectiveness of community-based health promotion projects, be they quantitative (numerical meta-analyses) or qualitative (narrative reviews), both have significant drawbacks. Out of the work conducted for two initiatives, the developments for the Global programme on health promotion effectiveness carried at by the North American Region out of the International Union of Health Promotion and Education (IUHPE), as well as the work conducted for the ECIP (effectiveness of community interventions project) of Health Canada, a new way to approach the issue of effectiveness is proposed. Based on a "realist synthesis" epistemological position, this approach has led us to the first formulation of a framework aiming at identifying the mechanisms that explain why local programs are successful. PMID- 15551691 TI - [Preffi 2.0: a Dutch instrument to analyse the effectiveness of health promotion interventions]. AB - Preffi is a set of guidelines with items relevant to the effectiveness of health promotion projects, reflecting scientific and practical knowledge. Preffi is used to assess quality at various stages of an intervention, either to critically evaluate one's own project or to comment on projects proposed by others. This article describes the Preffi model, its scoring method and the systematic, seven step development process that led to its second version: Preffi 2.0. The draft version of Preffi 2.0 was tested for usefulness among 35 experienced Health Promotion specialists. They gave the instrument an average overall score of 7.7 on a scale of 10. The large majority of them evaluated it as valuable, complete, clear, well-organised and innovative. Users have commented that applying Preffi to a project yields a balanced and useful assessment, as well as a clear overview of points in the project that could be improved. PMID- 15551692 TI - [Measuring the effectiveness of health promotion policy: what can be learnt from Australian success stories?]. AB - In 1987, the Australian state of Victoria has created VicHealth, the Victorian Health Promotion Foundation, which has since been quite successful in implementing effective health promotion policies. This paper, after presenting briefly three of these success stories (tobacco control, road trauma decrease and skin cancer prevention), goes on to derive from them the essential steps of successful health promotion interventions. It concludes by reflecting on the nature of the intelligence gathering systems required to anchor good health promotion in good science, and on the barriers to proper measurement that are still pervasive. PMID- 15551693 TI - [The International Union for Health Promotion and Education's Global Programme on Health Promotion Effectiveness]. AB - The Global Programme on Health Promotion Effectiveness (GPHPE) is a multi-partner initiative, coordinated by the International Union for Health Promotion and Education (IUHPE) in collaboration with the World Health Organization, and supported by a number of partners from across the world. The GPHPE aims to raise standards of health promoting policy-making and practice worldwide by: reviewing evidence of effectiveness in terms of health, social, economic and political impact; translating evidence to policy makers, teachers, practitioners, researchers; stimulating debate on the nature of evidence of effectiveness. The principal challenge for the GPHPE is to develop a sustainable approach with adaptations suitable to different regional needs, whilst maintaining the high quality for which the European work that triggered the programme is recognised. Although varied, the regional activities are all clearly designed to contribute to the global body of knowledge that the GPHPE aims to build. The article presents the origins of the GPHPE, its operational structure and zooms in on some of the specific activities of the European Region. PMID- 15551694 TI - [The effectiveness of health promotion in France: comments from a round table of French experts]. AB - In order to comment on the situation in France, a panel of six experts originating from the academic, the governmental and the service providing worlds responded to two questions, in a format that left room for significant interaction with the participants. The first question was: According to your own experience, please identify one health promotion or health education initiative that you consider effective. The second: If there was one priority to address in relation to the issue of health promotion effectiveness in France, what would that be? This paper summarises the key elements of the experts' answers to each question. PMID- 15551695 TI - [Evaluation of social behavior in health education interventions: why is France lagging behind?]. AB - In this paper, after differentiating evaluation from classical research, three issues are addressed: How to deal with the tyranny of randomised controlled trials? How to ascribe behavioural effects to prevention or to treatment? How to disentangle the short and long term effects of interventions? It is argued that addressing these three issues would help to understand why France, as compared to other Anglo-Saxon countries for instance, has been less concerned by the issues of evaluation and effectiveness, and where it could go if it wanted to get more involved around these matters. PMID- 15551696 TI - The big story. If you've goit it, use it--or are foundations too scared to use their borrowing freedoms? PMID- 15551697 TI - Joined-up working ideas. And one for all. PMID- 15551698 TI - Data briefing. Mapping needs. PMID- 15551699 TI - Clinical management where medicine meets management. Evening the odds. PMID- 15551700 TI - A year in Bristol. Shipshape--Bristol fashion. AB - The image of United Bristol Healthcare trust is changing; its rating has risen from zero stars to two. The new management team is struggling with tight finances, but the trust must break even. UBHT managers are aiming to develop a more corporate culture. PMID- 15551701 TI - Finance. Save and prosper. PMID- 15551702 TI - Law. Live and let live. PMID- 15551703 TI - HSJ people. A sickie situation. PMID- 15551704 TI - Drivers and outcomes of PACS. PMID- 15551705 TI - Beyond clinical documentation: using the EMR as a quality tool. PMID- 15551706 TI - Works as advertised. PMID- 15551707 TI - Conversational computing. PMID- 15551708 TI - Strategic planning supports ED automation. PMID- 15551709 TI - Market competition gives way to benefits of cooperation. PMID- 15551711 TI - PACS/RIS imaging hotlist. PMID- 15551710 TI - Achieving transparency in the claims payment process. PMID- 15551712 TI - HSAs will catalyze adoption of EHRs. PMID- 15551713 TI - Too much information. PMID- 15551714 TI - What is your diagnosis? Solitary mastocytoma. PMID- 15551715 TI - Pyogenic granuloma. AB - Pyogenic granuloma (PG) is an acquired vascular lesion of the skin and mucous membranes common to the pediatric age group. PG appears as a solitary red nodule on the head or neck. The nodule is prone to hemorrhage, and bleeding is often refractory to pressure. The etiology of PG is unknown, but proposed agents include trauma, infection, and preceding dermatoses. Several surgical treatments are available with variable cosmetic results and recurrence rates. PMID- 15551716 TI - Atrophic dermatofibrosarcoma protuberans: a case report and reappraisal of the literature. AB - Dermatofibrosarcoma protuberans (DFSP) is an uncommon cutaneous malignancy. Unusual presentations described as atrophic have been documented. A case of DFSP with both clinical and histologic atrophy is presented, and all cases purporting atrophy with this tumor are reviewed. Meaningful trends are extracted from this data. In addition, the imprecise use of the term atrophic in regard to DFSP is clarified. We maintain that the variant of atrophic DFSP that mimics atrophoderma or anetoderma, as in this case, is the rarest variant of atrophic DFSP. Atrophic DFSP should be in the differential for depressed lesions on the trunks of women or on the lower extremities of children. PMID- 15551717 TI - A refractory fixed drug reaction to a dye used in an oral contraceptive. AB - A young woman presented with a classic fixed drug eruption (FDE) after taking the inactive green pills of her oral contraceptives (OCs). The patient's history was unique in that the FDE did not occur every time she took the inactive pills but was refractory, occurring every third month within hours after she took the green pills. After discontinuing the green pills but continuing the active oral contraceptive pills, the patient has not experienced a recurrence of the rash in more than 2 years. This case report reviews the unusual phenomenon of refractory periods in FDEs and highlights the importance of understanding this phenomenon in the diagnosis of drug eruptions. PMID- 15551719 TI - Leishmaniasis acquired in the Iraqi Theater of Operations: lessons learned. AB - Between August 2002 and February 2004, the Department of Defense identified 522 cases of cutaneous leishmaniasis in military personnel. This commentary examines reasons why there were so many cases of leishmaniasis during this conflict as compared with Operation Desert Storm. Lessons learned can be applied to reduce the risk to US personnel during future conflicts. Among the factors to be considered are environment, exposure, vector control, and the failure to deliver insect repellent to deployed US personnel. PMID- 15551718 TI - Treatment of recalcitrant pemphigus vulgaris with the tumor necrosis factor alpha antagonist etanercept. AB - The treatment of pemphigus vulgaris (PV) is generally regarded as challenging. Patients with the disease require long-term systemic therapy, creating concern for the toxicities of these treatments. Corticosteroids, as drugs of first choice, often must be combined with steroid-sparing agents to prevent hazardous long-term side effects. We describe a 62-year-old woman with long-standing PV whose cutaneous disease responded to therapy with the tumor necrosis factor alpha (TNF-alpha) antagonist etanercept, which was started for treatment of her inflammatory seronegative arthritis. To our knowledge, this is the first report of its efficacy in the treatment of PV. PMID- 15551720 TI - Noxious sensory perceptions in patients with mild to moderate rosacea treated with azelaic acid 15% gel. AB - Patients with rosacea form a unique subset of the sensitive skin population because of the barrier defects inherent in this condition and the increased propensity for burning/stinging from topical products. This propensity for burning/ stinging when medications, skin care products, or cosmetics are applied to the facial skin has been frequently documented but never quantified. The objective of this 2-week study was to determine the prevalence of heightened neurosensory perceptions of burning/stinging in a random population of 40 women with mild to moderate rosacea defined as 15 or fewer inflammatory papules or pustules. Also evaluated was the effect of azelaic acid 15% gel on barrier function and facial stinging utilizing transepidermal water loss (TEWL), corneometry, and lactic acid facial sting tests as noninvasive measurement criteria. At baseline, the incidence of lactic acid stinging among these rosacea subjects was 62.5%, which is substantially higher than observed in the general population. Two weeks after application of azelaic acid 15% gel, no evidence of barrier damage was noted on TEWL or corneometry tests. Moreover, there was no statistical relationship between lactic acid stinging and a stinging response that is occasionally reported with exposure to azelaic acid 15% gel. PMID- 15551721 TI - Safety and efficacy of combined use of 4-hydroxyanisole (mequinol) 2%/tretinoin 0.01% solution and sunscreen in solar lentigines. AB - The objective of this open-label, noncontrolled study was to evaluate the safety of a combination solution containing 4-hydroxyanisole (mequinol) 2%/tretinoin 0.01% (Solage) with a sunscreen in the treatment of solar lentigines. The study included a total of 406 subjects for a treatment period up to 24 weeks. Efficacy was evaluated clinically by grading the pigmentation level of the treated areas on the face and forearms. A total of 378 subjects were included in the safety population. Of the 173 subjects with skin-related and treatment-related adverse events, severity was reported as mild in 79 subjects, moderate in 71, and severe in 23. Hypopigmentation was observed in 4 subjects and had definitively resolved in 3 of these subjects at the end of the study or after treatment had been discontinued. Halo hypopigmentation was reported in 16 subjects. No allergic reactions were observed. Efficacy evaluation was based on data for 370 subjects. A total of 325 (88%) subjects had facial target lesions almost clear to clear, and a total of 298 (81%) subjects had forearm target lesions almost clear to clear. Our study shows that the mequinol 2%/tretinoin 0.01% solution is effective, convenient, and safe in the treatment of solar lentigines. PMID- 15551722 TI - The protocols of the elders of dermatology. PMID- 15551723 TI - Assessing the prevalence of depression in Punjabi and English primary care attenders: the role of culture, physical illness and somatic symptoms. AB - Previous studies exploring the prevalence of depression among South Asians reported inconsistent findings. Research artefacts due to sampling bias, measurements errors and a failure to include ethnographic methods may all explain this. We estimated the prevalence of depression, and variations of prevalence with culture, cultural adaptation, somatic symptoms and physical disability in a cross-sectional primary care survey of Punjabi and English attendees. We included a culture specific screening instrument, culturally adapted the instruments and offered bilingual interviews. We found that, compared with their English counterparts, depressive diagnoses were more common among Punjabis, Punjabi women, Punjabis with physical complaints and, contrary to expectation, even Punjabis with low scores for somatic symptoms. PMID- 15551724 TI - Do South African former detainees experience post-traumatic stress? Circumventing the demand characteristics of psychological assessment. AB - Most research on persons subjected to physical or psychological torture for political reasons has framed this experience as traumatic, with the sequelae approximating the diagnostic criteria of post-traumatic stress disorder (PTSD). Yet, critiques of the trauma model have called attention to the fact that PTSD represents a Western conceptualization of the concerns of persons who have survived stressful experiences. In order to determine whether symptoms of traumatization are salient psychiatric phenomena for South African former detainees, semi-structured qualitative interviews were conducted with 20 respondents who were detained and tortured for political reasons during the apartheid era. Interviews were transcribed and analysed for thematic content using a grounded theory approach. Results showed that although the main concerns expressed were unrelated to traumatization, participants also indicated that they experienced symptoms of post-traumatic stress. These data suggest that although too great a focus on traumatic responses may be misplaced, it remains important to consider the possibility that former detainees may exhibit symptoms of this nature. Consequently, critiques of the trauma discourse as a Western phenomenon need to be tempered with evidence of the lived reality of psychological sequelae experienced by this population. PMID- 15551725 TI - Authority, dialogue and polyphony in psychiatric consultations: a Latvian case study. AB - This article explores the impact of economic liberalization on psychiatric theory and practice in Latvia. Through a close reading of four psychiatric consultations it examines the way in which contradictions between a market economy, its attendant philosophy of unlimited individual opportunity and the implementation of these ideas in the lives of individuals are negotiated by psychiatrists and their patients. It examines the uses of psychological and somatic vocabularies and questions their claimed connections to an enlarged sense of agency or its denial. PMID- 15551726 TI - Sell a book and cook a dog: misery, memory and space from Siberian camps to Apartheid. AB - In this article, the life story of a Polish woman in South Africa is used to illustrate that stories of exile are 'matter out of place' in host countries. Space is a central category in both the story of exiles and in the collective history of receiving countries, but it has different emotional, social, cultural and political meanings. Sometimes stories of exile are denied a place in the social memory of a society. Therefore, recollection and making sense of experienced misery in foreign contexts is often a difficult and lonely undertaking. The article links the story to macro-political processes and attempts to answer questions regarding the consequences of the lack of meaningful space for exiles to remember. Collective and national memories and ceremonies are often exclusive and offer no place for 'alien' memories. Therapeutic repertoires often fail to address the emotional value of communal sharing a socio-cultural and (meta) physical space. It is argued that because the state does not take responsibility for providing a (symbolic) space for the mourning and reconciliation of people whose memories are out of place, transcultural psychiatry - its subsystem - will have to reflect on how to provide such a space. PMID- 15551727 TI - A psychiatrist in cultural transition: personal and professional dilemmas. AB - This article describes the author's personal and professional dilemmas, encountered as a psychiatrist in the process of migration from Central Europe to New Zealand. The dilemmas include: (i) personal experiences in the stages of migration, (ii) struggles within the author's own multi-ethnic community, (iii) biculturalism, (iv) the unexpected seriousness of psychopathology, and (v) the tendency for overidentification. Professionals in a cultural transition should be aware that personal and professional challenges are interconnected. The concepts of loyalty, relational ethics and existential humanism were valuable in the resolution of these dilemmas. PMID- 15551728 TI - Medical compliance in the East. PMID- 15551729 TI - Neuroendocrine cell markers for pancreatic islets and tumors. AB - The authors review the application of a variety of neuroendocrine cell markers to identify pancreatic islet cells and tumors. In the past, several empiric histochemical techniques had been used to demonstrate neuroendocrine cells, particularly the Grimelius argyrophilic stain. The development of immunohistochemistry made it possible to demonstrate specific cell products such as regulatory peptides, thus allowing the classification of pancreatic neuroendocrine tumors with a view to clinical symptoms. However, it is not always possible to visualize regulatory peptides in these tumors. It is therefore important to use broad-spectrum neuroendocrine cell markers to identify the neuroendocrine nature. These markers are proteins localized in the secretory granules (core- or membrane-related), in the cytosol, or in the cellular membrane. The markers most commonly used in routine histopathology are the secretory granule proteins chromogranin A and synaptophysin and the cytosolic enzyme neuronspecific enolase. Other new markers (e.g., synaptic vesicle protein 2) are of general diagnostic value. Region-specific antibodies to chromogranin A can be valuable in differentiating between benign and malignant neuroendocrine tumors. Some markers may be related to the functioning characteristics of pancreatic neuroendocrine tumors, such as prohormone convertases. In addition, markers giving further complementary information have been identified, such as five somatostatin receptor subtypes, the expression of which varies markedly in pancreatic neuroendocrine tumors. Antibodies against all somatostatin receptor subtypes are now commercially available, and immunohistochemical investigation of its expression should be routinely applied when considering treatment with somatostatin analogs. PMID- 15551730 TI - Immunohistochemical analysis of biliary tract lesions. AB - The distinction among inflammatory, benign, and malignant lesions of the biliary tract can at times be difficult. Several methods have been used, including immunohistochemistry (IHC), with variable success. We evaluated a panel of IHC stains to determine their utility in discriminating between bile duct lesions. Formalin-fixed, paraffin-embedded 4-microm sections from 12 inflammatory lesions, 10 bile duct adenomas, and 13 bile duct carcinomas were immunostained using a modified avidin-biotin-complex technique after epitope enhancement using antibodies for p53, Ki-67, and bcl-2. For p53 and bcl-2, greater than 1% of cells staining positive was interpreted as positive. The proliferation index was calculated by determining the number of Ki-67-positive cells in a 1000 cell count. In the inflammatory group, 0 of 12 reacted with anti-p53, 2 of 12 were positive with anti-bcl-2, and the proliferation index with was 22.9% +/- 3.9%. Two of 10 bile duct adenomas showed reactivity with anti-bcl-2, and none were decorated with anti-p53 or Ki-67. In the carcinoma group, 6 of 13 were positive with anti-p53, 9 of 12 were positive with anti-bcl-2, and the proliferation index was 35.3% +/- 5.5%. The proliferation rates differed significantly between groups (P < 0.05). The presence of bcl-2 and p53 immunoreactivity coupled with a high proliferative rate in a biliary tract lesion suggests a malignant process. A panel using these antibodies may be useful in difficult cases. PMID- 15551731 TI - Immunohistologic parameters in minimal morphologic change duodenal biopsies from patients with clinically suspected gluten-sensitive enteropathy. AB - Subclinical or latent cases of gluten-sensitive enteropathy (GSE) are difficult to diagnose, and serology-positive, histology-negative (minimal morphologic change) and serology-negative, histology-positive patients have been identified. Both, particularly the histology-negative group, require the correct diagnosis for proper management, especially because the concept of minimal histologic change GSE has escaped attention in standard textbooks. We assessed the numbers and distribution of intraepithelial T cells and their subsets with CD3, CD8, and CD4 immunostaining and examined for crypt hyperplasia with mitotic and Ki-67 proliferative indices with the aim of refining the criteria for the diagnosis of minimal change GSE. Duodenal biopsies from 46 clinically suspected cases of GSE tested for antigliadin, antiendomysium, and antitissue transglutaminase antibodies were divided into four groups: serology-positive, histology-positive (S+H+, n = 20); serology-positive, histology-negative (S+H-, n = 22), representing the minimal morphologic change group; serology-negative, histology positive (S-H+, n = 4); and serology-negative, histology-negative (S-H-, n = 28), controls with histologically normal duodenal biopsies obtained for unrelated reasons. The numbers of CD3+ and CD8+ intraepithelial T cells (IETCs) were significantly higher in histology-positive biopsies with (mean, 40.3/100 and 39.3/100 enterocytes, respectively) and without positive serology (mean, 33.3/100 and 35/100 enterocytes, respectively) compared with all other groups (S+H-, mean, 26.5/100 and 24.3/100 enterocytes, respectively; S-H-, mean, 23.3/100 and 17.9/100 enterocytes, respectively). Values for Ki-67 index in crypt enterocytes were also significantly different between histology-positive and histology negative groups (P = 0.000). The distribution of CD3+ and CD8+ IETCs was mostly even along the surface enterocytes in histology-positive cases compared with the controls, which showed an uneven distribution. The 2 parameters that significantly discriminated between minimal morphologic change GSE (S+H-) and controls (S-H-) were Ki-67 index (P = 0.007) and the distribution pattern of CD8+ IETCs (P = 0.049). CD4 IETC counts were generally low, with no significant difference between all groups. The few S-H+ cases seen most likely represented false-negative serology, because the assessed parameters of this group and S+H+ cases were indistinguishable. PMID- 15551732 TI - Reduced PTEN expression is associated with poor outcome and angiogenesis in invasive ductal carcinoma of the breast. AB - Loss of PTEN expression has been associated with advanced stages of tumor. Tumor angiogenesis is involved in tumor progression. In breast cancer, a high frequency of mutations of the PTEN locus has been reported. However, the prognostic importance of PTEN expression and its correlation with angiogenesis in breast cancer have not been well established. Formalin-fixed, paraffin-embedded tissues from 99 women with a primary diagnosis of invasive ductal carcinoma were evaluated for PTEN expression by immunohistochemical methods. The microvessel density (MVD) was also studied by immunohistochemical labeling of endothelial cells with CD34 antibody. Computerized image analysis was used to evaluate MVD. Reduced PTEN expression was seen in 27.3% of invasive ductal carcinoma. The MVD ranged from 22.0 to 197.0, with a median value of 58.5 (65.4 +/- 27.9). Reduced PTEN expression correlated with lymph node status (P < 0.01), tumor grade (P < 0.05), and tumor-node-metastasis (TNM) stage (P < 0.05). There was a statistically significant correlation between reduced PTEN expression and increased MVD (P < 0.05). The mean MVD was higher in reduced PTEN-expressive tumors, irrespective of stage, compared with normal PTEN-expressive tumors with the same stage. On multivariate analysis, only TNM stage and reduced PTEN expression correlated with survival. Our results suggest that reduced PTEN expression may be an independent prognostic indicator in patients with invasive ductal carcinoma. PTEN loss may be associated with increased tumor angiogenesis. PMID- 15551733 TI - Differential expression patterns of c-REL protein in classic and nodular lymphocyte predominant Hodgkin lymphoma. AB - Classic Hodgkin lymphoma (cHL) is characterized by numerical gains of the short arm of chromosome 2. The high frequency of 2p overrepresentation including REL, particularly in the nodular sclerosis subtype suggests that constitutive activation of nuclear factor kappaB/REL is a hallmark of Reed-Sternberg (RS) cells. The aim of this study was to investigate c-Rel protein expression patterns in cHL and nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) cases by immunohistochemical analysis. A total of 79 cases of HL were analyzed, which included 59 cases of cHL (49 nodular sclerosis; 8 mixed cellularity; 2 lymphocyte rich) and 20 cases of NLPHL. Positive staining was defined in this study as a reaction seen in the nuclei or nuclei and cytoplasm of RS or lymphocytic and histiocytic (L&H) cells in cHL and NLPHL cases, respectively. The percent positivity of c-REL staining of RS cells in cHL was seen in 51 of 59 cases (86.4%). No significant difference in c-REL expression was seen between nodular sclerosis (42 of 49, 85.7%) and mixed cellularity subtypes (7 of 8 cases, 87.5%; P = 1). In comparison, positive c-REL protein expression in L&H cells was seen in 5 of 20 NLPHL cases (25.0%). Therefore, significantly higher positivity of RS cells in cHL was seen compared with positivity of L&H cells in NLPHL; 86.4% vs. 25.0%; P = 0). Expression of Epstein-Barr virus latent membrane protein was seen in 6 of 30 cases (19.0%; 25 cHL, 5 NLPHL) and EBER1 in 5 of 27 cases (18.5%; 24 cHL, 3 NLPHL). The presence of Epstein-Barr virus did not correlate with c-REL protein expression (P = 1). Our results demonstrate that there is differential c REL protein expression in cHL in comparison with NLPHL and suggest that c-REL may play a role in the pathogenesis of classic Hodgkin lymphoma. PMID- 15551734 TI - Cutaneous immunocytoma: a clinical, histologic, and phenotypic study of 11 cases. AB - Immunocytomas represent low grade B cell lymphomas related to marginal zone lymphoma but with a predominance of cells having plasmacytic features. Eleven patients presented with lesions compatible with primary cutaneous immunocytoma. The expression of CD2, CD3, CD5, CD20, CD21, CD23, CD43, CD56, CD79, and bcl-2 was analyzed immunohistochemically and of lambda and kappa light chains by an in situ hybridization assay. There were 6 men and 5 women ranging in age from 43 to 76 years. The most common clinical presentation was as extremity based clustered erythematous brown papules. Therapy with local irradiation or Rituximab resulted in lesional resolution. Underlying illnesses included Sjogren's syndrome, hepatitis C, ulcerative colitis, autoimmune thyroid disease, and rheumatoid arthritis. Four patients were taking medications previously associated with immune dysregulation. In two patients in whom a paraproteinemia was uncovered. The most common pattern light microscopically was perivascular small lymphocytic and plasmacellular infiltrates mimicking architecturally a reactive process. Phenotypic studies revealed a marginal zone (MZL) phenotype amid the small atypical lymphocytic infiltrate and highlighted a reactive background population of non-neoplastic T and B cells; light chain restriction was seen amid the plasma cells. In one case there was EBER staining of plasma cells while in another case in whom there was hepatitis C seropositivity staining of plasma cells for hepatitis C associated RNA transcripts was observed. Primary cutaneous immunocytoma appears to arise from a pre-existing state of reactive lymphoid hyperplasia. latrogenic and endogenous immune dysregulation including in the context of lymphotropic viral infections is implicated. PMID- 15551735 TI - P53 protein is a reliable marker in identification of renal tubular injury. AB - Acute tubular necrosis (ATN) ranges from minimal histologic changes to overt necrotic tubules. Although histologic changes on routine stained sections can be seen in many ATN cases, they may be subtle in some cases. In some cases, electron microscopy may reveal more reliable findings to support a diagnosis of ATN. Thus, a molecular marker to confirm acute tubular damage and to differentiate mild from moderate tubular injury could provide more reliable detection of ATN at the light microscopic level. In this study, sections from native and transplant renal biopsies with the diagnosis of ATN were stained immunohistochemically for p53, an upstream marker for DNA damage, and compared with donor baseline biopsies as controls. The transplant and native ATN kidney groups had significantly higher numbers of p53 nuclear staining in renal tubular epithelium (transplant ATN: 4.58 +/- 1.51/mm2, n = 18, and native ATN: 6.12 +/- 1.99/mm2, n = 13) than the donor baseline group (1.09 +/- 0.51/mm2, n = 16) or controls-normal renal parenchyma away from tumors (0.029 +/- 0.017). Cases with moderate ATN changes showed significantly increased p53 tubular staining (transplant ATN: 9.20 +/- 2.59/mm2, n = 8, and native ATN: 14.3 +/- 1.88/mm2, n = 5) when compared with the mild ATN cases (transplant ATN: 0.87 +/- 0.30/mm2, n = 10, and native ATN: 1.01 +/- 0.39/mm2; n = 8). In summary, there was direct correlation between nuclear p53 staining and morphologic changes seen microscopically and ultrastructurally, suggesting that p53 can be used as a reliable marker of cellular damage to aid in the diagnosis of ATN. PMID- 15551736 TI - Enterogenous cysts of the spinal canal and cerebellopontine angle. AB - Enterogenous cysts are rare, benign lesions involving the spinal canal or the cerebellopontine angle. Typically they present with compression of the spinal cord or cranial nerves. They are usually 1 cm or less in size and are typically lined by columnar mucin-producing epithelium resembling enteric epithelium or columnar ciliated epithelium resembling respiratory epithelium. The case records of the Division of Neuropathology at Indiana University were reviewed for cystic lesions of the spinal canal and cranial cavity for a 26-year period. Seven surgically resected enterogenous cysts were identified. The original slides were studied. Additional sections were stained with mucicarmine, and immunohistochemical stains including thyroid transcription factor 1, epithelial membrane antigen, and cytokeratin 5/6 (CK5/6) were performed. Electron microscopy was performed in 1 case. The cysts ranged in size from 4 to 10 mm. One had stratified cuboidal epithelium, 1 had ciliated columnar and stratified squamous epithelium, 1 had columnar mucinous epithelium, and 4 had ciliated columnar epithelium. Epithelial membrane antigen and CK5/6 positivity were observed in all cases. Strong CK5/6 positivity was seen in the basal cells, with little or no immunoreactivity in the ciliated cells. Mucicarmine positivity was observed in 4 cases, and thyroid transcription factor 1 positivity was observed in 2 cases. Electron microscopy demonstrated well-developed stereocilia, distinct basal cells, and a thin basement membrane. The histopathologic, ultrastructural, and immunologic findings in these lesions are similar to those present in the bronchial epithelium, supporting the hypothesis of endodermal origin for these lesions. PMID- 15551737 TI - Overexpression of neural cell adhesion molecule in regenerative muscle fibers in 3-hydroxy-3-methylglutaryl coenzyme: A reductase inhibitor-induced rhabdomyolysis. AB - Skeletal muscle degeneration is a side effect of cholesterol-lowering hydroxymethylglutaryl coenzyme A reductase inhibitors. The expression of the cell cell adhesion proteins, neural cell adhesion molecule and neural-cadherin was studied in a case of rhabdomyolysis induced by the hydroxymethylglutaryl coenzyme A reductase inhibitor cerivastatin. Neural cell adhesion molecule and N-cadherin participate in the interactions of muscle cells during skeletal myogenesis. In the adult muscle, neural cell adhesion molecule is restricted to neuromuscular sites but is re-expressed in denervated muscle and in rhabdomyolysis. Our results show expression of neural cell adhesion molecule in regenerative skeletal muscle fibers but not in degenerated or unaffected fibers in cerivastatin-induced rhabdomyolysis. In contrast, N-cadherin was not expressed. The presence of apoptotic cells was studied by a fluorescence-based Tdt-mediated dUTP nick-end labeling in the same sections. Apoptosis was detected in degenerative fibers and inflammatory cells but not in regenerative fibers. We hypothesize that the expression of neural cell adhesion molecule in regenerative fibers may have a protective role against apoptosis during rhabdomyolysis. Cerivastatin-induced rhabdomyolysis appears to have common features with rhabdomyolysis of other causes. The immunohistochemical study of neural cell adhesion molecule can serve as an additional tool in the evaluation of muscle regeneration in rhabdomyolysis. PMID- 15551738 TI - P63 expression in lung carcinoma: a tissue microarray study of 408 cases. AB - p63 is a recently discovered member of the p53 family that has been shown to be important in the development of epithelial tissues. p63 may also play a role in squamous cell carcinomas of the lung, head and neck, and cervix, and its expression is increased in these tumors. The purpose of this study was to investigate the expression of p63 in a broad spectrum of histologic types of lung tumors. A total of 441 cases of primary lung tumors with follow-up data were identified, and the paraffin-embedded tissue blocks were used to construct a duplicate core tissue microarray. After review of the tissue cores, 408 cases, consisting of 123 squamous cell carcinomas, 93 adenocarcinomas, 68 large cell carcinomas, 68 classic carcinoids, 31 atypical carcinoids, 11 large cell neuroendocrine carcinomas, and 14 small cell carcinomas, were adequate for analysis. Immunohistochemistry was performed at 2 different laboratories using monoclonal antibody 4A4 to detect the expression of p63, using different staining protocols. p53 expression was also studied with immunohistochemistry using monoclonal antibody DO-7. Kaplan-Meier curves were plotted to compare the survival of p63-expressing versus nonexpressing tumors. A large proportion of squamous cell carcinomas expressed p63 (96.9%), most showing strong positive nuclear immunoreactivity. Expression in other nonsmall cell lung cancers was also present. Thirty percent of adenocarcinomas and 37% of large cell carcinomas showed p63 expression. In the neuroendocrine tumors, an increasing proportion of tumors stained for p63 as tumor grade increased; 1.9% of classic carcinoids, 30.8% of atypical carcinoids, 50% of large cell neuroendocrine carcinomas, and 76.9% of small cell carcinomas were positive. Approximately half of the positively staining neuroendocrine cases showed strong staining. Expression of p63 was of prognostic significance in neuroendocrine tumors (P < 0.0001), with higher-grade tumors more likely to express p63. Correlation between p63 and p53 expression was not observed (P = 0.18) in nonsmall cell lung cancer; however, a significant correlation between the 2 markers was found in neuroendocrine tumors (P < 0.0001). p63 staining was repeated with a different staining protocol, yielding similar results overall but a lower percentage of positive cases (34.2% vs. 48.4% of tumors positive). In conclusion, p63 expression is consistently expressed in squamous cell carcinoma in the lung, but is also expressed in a subset of adenocarcinomas and large cell carcinomas. Pulmonary neuroendocrine tumors also show p63 staining in some instances, particularly in higher-grade tumors, and the majority of small cell carcinomas are p63-positive. These results suggest that p63 may be involved in oncogenesis in a broader range of tumors than was previously thought. PMID- 15551739 TI - Her2 amplification: correlation of chromogenic in situ hybridization with immunohistochemistry and fluorescence in situ hybridization. AB - Detecting Her2 gene amplification has become routine in predicting therapeutic responsiveness in patients with breast carcinoma. Fluorescence in situ hybridization (FISH) is a common technique for detecting Her2 amplification, yet dark field fluorescence microscopy remains problematic for many pathologists. Thus, a technique such as chromogenic in situ hybridization (CISH), in which the more familiar light microscopy can be used, is appealing. Paraffin-embedded sections from 61 breast carcinomas were tested for Her2 amplification by immunohistochemistry (IHC) and CISH. FISH was used to confirm CISH results. Excellent correlation was found between IHC and CISH except in cases considered negative (1+ on the DAKO scale) by IHC. CISH detected low-level Her2 amplification in 4 of 9 of these cases. Amplification was subsequently confirmed by FISH in all but 1 case. When compared with FISH, CISH was more sensitive than IHC for detecting low levels of Her2 gene amplification. Moreover, excellent concordance was found between FISH and CISH, supporting the conclusion that the CISH assay for Her2 gene amplification provides an accurate, effective, and practical alternative to FISH. PMID- 15551740 TI - Determination of light chain restriction in fine-needle aspiration-type preparations of B-cell lymphomas by mRNA in situ hybridization. AB - The purpose of this study was to investigate whether B-cell clonality could be demonstrated in fine-needle aspiration (FNA)-type preparations by using automated and manual in situ hybridization (ISH) systems. FNA-like preparations were made from 10 cases of B-cell lymphoma and 5 cases of reactive lymphoid hyperplasia. Kappa/lambda expression was determined using an automated mRNA ISH assay or a manual ISH system. Other variables tested included type and length of fixation, protease digestion, and time in chromogen. Clonality data were corroborated by either flow cytometry or tissue-based analysis. Optimal conditions required formalin fixation, strong protease digestion, and prolonged hybridization and chromogen times; under these conditions, monoclonality was demonstrated by in situ in 8 of 10 cases. Each of the five cases of reactive lymphoid hyperplasia showed polyclonal light chain expression by automated mRNA ISH. In situ kappa/lambda mRNA analysis of FNA-type specimens allows direct determination of monoclonality in cytologic preparations. PMID- 15551741 TI - Fluorescence in situ hybridization on formalin-fixed and paraffin-embedded tissue: optimizing the method. AB - Fluorescence in situ hybridization (FISH) is widely used to study numerical and structural genetic abnormalities in both metaphase and interphase cells. The technique is based on the hybridization of labeled probes to complementary sequences in the DNA or RNA of the cells. Interphase FISH is most often applied on cytologic material such as hematologic smears or imprints, but the method is also used to study genetic changes in tissue sections when morphology is important or when cytologic material is not available. In cases in which the presence of intact nuclei is of importance, such as quantitation of signals as in triploidy, it is possible to isolate nuclei from paraffin-embedded tissue. However, using formalin-fixed paraffin-embedded tissue, either in thin sections or as isolated nuclei, one encounters a range of technical problems, paralleling those met in immunohistochemistry. Variations in time lapse between removal of tissue and fixation, duration of fixation, enzymatic pretreatment, hybridization conditions, and posthybridization washing conditions are important factors in the hybridization. In this study, we have listed the results of a systematic approach to improve FISH on isolated nuclei and tissue sections from formalin-fixed, paraffin-embedded tissue. PMID- 15551742 TI - Direct and repeat uses of tissue sections as templates for liquid-phase polymerase chain reaction amplification: applications and implications. AB - The liquid-phase polymerase chain reaction (PCR) technique is the most commonly used method for the amplification of genetic materials, although it requires the lysis of cells for DNA or RNA extraction, making it impossible to visualize the distribution and subcellular localization of the biomolecules. This study intended to assess whether tissue sections may be directly and repeatedly used as templates for liquid-phase PCR amplifications. Consecutive paraffin sections of breast tissues were placed on gamma-methacryloxypropyltrimethoxysilane-coated microscopic cover glasses perforated with a diamond knife into strips. After hematoxylin and eosin or immuno-staining, the strip of interest was inserted into a PCR tube for amplifications, and the adjacent strip with the same tissue was subject to microdissection, DNA extraction, and PCR amplifications. To use the strip repeatedly, it was transferred into a new PCR tube and amplified with a new primer set, after an initial amplification for 5 to 7 cycles. Then, initially amplified samples were amplified to a total of 40 cycles. An equal volume of PCR products from the strip and DNA extract were loaded side by side for electrophoresis and detection. The strip and DNA extract from the same tissue yielded a very comparable quality and quantity of PCR products with the same primer sets. The strip, however, could be repeatedly used for PCR amplifications with substantially more primer sets. In addition, the strip could be used for immunohistochemical or other molecular assays after PCR amplifications. Further studies with the same protocol on strips containing chromosomal spreads generated similar results. PMID- 15551743 TI - Mixed carcinoid and adenocarcinoma of the appendix: report of 4 cases with immunohistochemical studies and a review of the literature. AB - Four cases of mixed carcinoid and adenocarcinoma of the appendix were reported. All cases presented with a dominant cecal-appendiceal tumor mass and local metastasis. Two patients had multiple peritoneal implants mimicking primary peritoneal serous adenocarcinoma or carcinomatosis. Histopathologic features of the tumors are similar, with infiltrating microglandular and cribriform patterns of tumor nests, and variable numbers of goblet cells. A literature review of "goblet cell carcinoid" that included nonlocalized cases revealed a significant percentage (>14%) of tumor-associated death, in contrast to the classic carcinoid tumor. Immunohistochemical stains were helpful to separate these tumors from carcinoid tumors and primary peritoneal serous adenocarcinoma. Mixed carcinoid and adenocarcinomas were cytokeratin (CK)-20 positive, and negative or weakly positive for chromogranin A and synaptophysin. Carcinoid tumors were CK20 negative and diffusely positive for chromogranin A and synaptophysin. Peritoneal serous adenocarcinomas were CK20 negative. These cases were clinically aggressive, and 1 patient had multiple recurrences and responded partially to chemotherapy. PMID- 15551744 TI - Angiomyolipoma of the kidney: expanding disease spectrum demonstrated by 3 cases. AB - We report 3 recent cases of angiomyolipoma of the kidney. Although generally regarded as a benign neoplasm, angiomyolipoma rarely behaves in an aggressive manner, producing complicated clinical courses leading to metastasis and death. The presence of epithelioid elements within the tumor can result in difficulty differentiating benign from malignant angiomyolipoma and differentiating this tumor from renal adenocarcinoma. The presence of lymph node involvement can cause difficulty in differentiating multicentric disease in lymph nodes from metastasis to lymph nodes. The presence of cytologic abnormalities in the primary tumor can result in difficulty in differentiating atypia in benign angiomyolipoma from malignant sarcomatous transformation of a benign lesion. The 3 cases reported show many of these problems. Criteria for predicting malignancy in epithelioid tumors and sarcomatous transformation are not well recognized because of the rarity of this entity. The typical immunophenotype of all types of angiomyolipoma (cytokeratin-negative and melanomarkers-positive) is very useful in diagnosis but does not help in the differentiation from renal adenocarcinoma at frozen section. We report the empiric use of Ki67 and p53 in these cases as adjuncts to clinical and histologic assessment in predicting behavior. High Ki67 expression was a feature of malignant epithelioid angiomyolipoma. Low levels of p53 expression were seen in the angiomyolipoma with sarcomatous transformation. Benign angiomyolipomas were consistently negative for both Ki67 and p53. PMID- 15551745 TI - Electrocavernosogram in erectile dysfunction: a diagnostic tool. AB - Corpora cavernosa (CC) evoke electric waves that appear to be of diagnostic significance in evaluation of erectile dysfunction (ED). We investigated the hypothesis that electrocavernosography (ECG) exhibits different patterns in the various types of ED: neurogenic, vasculogenic, and psychogenic. Electrocavernosography was performed in the flaccid phase in 16 neurogenic, 28 vasculogenic (15 arteriogenic, 13 venogenic), and 24 psychogenic patients with ED, and in 15 healthy volunteers (controls). Two needle electrodes were introduced into the CC and the EMG activity was recorded in each of the 2 CCs of the same subject. Two 20 minute recording sessions were performed for each subject. The controls recorded slow waves (SWs) with regular rhythm and identical frequency, amplitude and conduction velocity from the 2 electrodes of the same subject. Random action potentials (APs) were superimposed on or followed the SWs. The ECG in the neurogenic ED recorded no waves in 14/16 patients and occasional irregular waves in 2. The SWs of the arteriogenic ED had irregular rhythm and variable and low parameters compared to those of the controls. The ECG of the patients with venogenic ED was similar to that of the controls, while the ECG of the psychogenic ED exhibited SWs with irregular rhythm and higher parameters than the controls. The study has revealed various ECG patterns in ED: "silent" in neurogenic. "bradyarrhythmic" in arteriogenic, "normal" in venogenic, and "overactive" in psychogenic ED. We suggest that electrocavernosography has the potential to function as an investigative tool in diagnosing the type of ED provided further studies are performed to verify the described findings. PMID- 15551746 TI - Fluorescent berberine binding as a marker of internal glycosaminoglycans sulfate in bovine oocytes and sperm cells. AB - The use of berberine as a biological marker of glycosamineglycans sulfate was employed to corroborate the presence of heparin in mammalian oocytes and sperm and its distribution in all the structures, or only in some specialized zones, of the male and female gametes. Oocytes and sperms were treated with 1.8 mM berberine for the presence of heparin and examined 10, 30, 60, and 120 minutes later. We have found that heparin is homogeneously distributed in all the zones of bovine oocytes and in sperm cells. When sperm cells are first treated with 80 microM of heparin and then berberine, 40% of them display in their post acrosomal region an intense yellow fluorescence. This may be in relation to the high amount of heparin binding sites due to the presence of the reticular membranous like system in this sperm region and in its possible role whereby gametes recognize and adhere to one another. Therefore, the use of berberine as a fluorescent marker of heparin represents clear proof of the presence of GAGs and their binding sites in the outside and inside of mammalian gametes, reinforcing the importance they play in the events of the process of fertilization. PMID- 15551747 TI - Probability based diagnostic biopsy specimens as predictors of tumor grade and stage found. AB - The Gleason score of prostatic adenocarcinoma in biopsy specimens was compared with the Gleason score of corresponding radical prostatectomy specimens from 78 patients with localized prostate cancer. Grading errors were found to be significant for well-differentiated (Gleason score 2-4) tumors. The accuracy was 6 (23%) for Gleason scores of 2-4 on needle biopsy. All of the Gleason scores of 8-10 on needle biopsy were graded correctly. When the preoperative Gleason score was <7, 20 (37%) patients had organ-confined lesions, while when preoperative Gleason score > or = 7, 5 (21%) patients were confined to the prostate. Discrepancies between the Gleason score of the biopsy material and prostatectomy specimens were larger for biopsy specimens with low Gleason scores than for biopsy specimens with high Gleason scores. Large differences existed between the Gleason histologic scores of the biopsy and prostatectomy specimens when only a single microscopic focus of the tumor in the biopsy specimen is low grade. Consequently, when tumor grade influences the clinical management of prostate cancer, patients with limited biopsy material, provided this material is not poorly differentiated, should probably undergo repeated biopsy to reduce the likihood of tumor sampling error. This awareness influences treatment policy, particularly for the watchful waiting criteria of prostate cancer. PMID- 15551748 TI - Differential regulation of inhibin subunits by germ cells in human testes. AB - Inhibin B is comprised of two dissimilar disulfide-linked subunits, termed alpha and betaB, and is physiologically more important than inhibin A in the male. The aim of this study was to investigate testicular expression of inhibin subtypes in infertile men to uncover any interaction between Sertoli cells and germ cells. Ten azoospermic patients with Sertoli cell only syndrome (SCO) and 39 oligozoospermic men were included in this study. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone concentrations were determined by chemiluminescence assays. The serum concentrations of inhibin B were measured by enzyme-linked immunosorbent assay. Immunohistochemical staining for the alpha subunit, betaA-subunit, and betaB-subunit of inhibin were performed on testicular biopsy specimens. The results were that serum inhibin B was undetectable in azoospermic men with SCO, while it was 133.8 +/- 82.0 pg/ml in oligozoospermic men. There was little expression of betaA in the testes of any patient. Expression of inhibin alpha and betaB was observed in Sertoli cells. The percentage of Sertoli cells expressing inhibin alpha was similar in azoospermic patients with SCO (55.3% +/- 20.6%) and in oligozoospermic patients (42.8% +/- 30.4%). In contrast, expression of betaB in Sertoli cells of azoospermic patients (24.9% +/- 16.8%) was lower than in oligozoospermic men (43.4% +/- 25.5%: P = 0.0308). There are no significant correlations between testicular expression of inhibin betaB and the serum inhibin B concentrations. The expression of inhibin betaB by Sertoli cells is dependent on the coexistence of spermatogenic activity within these seminiferous tubules, explaining why the level of inhibin B is low in patients with SCO. PMID- 15551749 TI - Prolonged and transient neonatal hypothyroidism on Leydig cell differentiation in the postnatal rat testis. AB - Hypothyroidism arrests the differentiation of adult Leydig cells (ALC) in the neonatal rat testis, and transient neonatal hypothyroidism produces a two-fold increase in the ALC numbers in the adult rat testis. We investigated 1) whether prolonged hypothyroidism beyond the neonatal period could continue to arrest the differentiation of the ALC, and 2) to understand how a two-fold increase in the number of ALC is produced in adult rats subjected to transient neonatal hypothyroidism. Three groups of Sprague Dawley rats were used; control, PTU-water group (transiently hypothyroid; added 0.1% propyl thiouracil/PTU to drinking water of lactating mothers at parturition until weaning of pups at day 21, pups were fed regular water thereafter), and PTU group (prolonged hypothyroid; mothers were fed 0.1% PTU in drinking water from parturition until pups were sacrificed at days 28 and 40 (pups had access to solid food after 21 days). Findings showed that PTU treatment continued to arrest ALC differentiation. Withdrawal of the PTU treatment at 21 days resulted in ALC differentiation by two-fold in number in PTU water rats. Findings on luteinizing hormone (LH)-stimulated androgen secretory capacity per testis in vitro agreed with the morphological data. These results confirmed that 1) thyroid hormone is crucial to the onset of ALC differentiation in the postnatal rat testis, 2) increased numbers of mesenchymal cells present in the hypothyroid testes differentiate into ALC upon withdrawal of the PTU treatment to produce a two-fold number of ALC in adult rats subjected to transient neonatal hypothyroidism (i.e., PTU-water treatment), and 3) numbers of ALC and mesenchymal cells increase with age at a rate of 2:1 during the process of ALC differentiation in testes of control and PTU-water rats. PMID- 15551750 TI - Ultrastructural morphology and morphometry of epididymal sperm in the volcano rabbit (Romerolagus diazi). AB - Sperm characteristics of Romerolagus diazi, an endemic endangered rabbit from Mexico's Higlands, are poorly known. Knowledge of gamete characteristics are urged for any conservation-oriented strategy and morphometry-based taxonomical database. Sperm lagomorph comparisons have been made at light microscopy resolution. Our goal was to analyze the ultrastructure of the R. diazi male gamete. Two wild animals were kept in captivity and the epididymus were obtained. Fixed gametes show a characteristic spatula-like morphology with a dilated forefront. The nucleus has an arrow head morphology lightly thicker at the base. Tail ultrastructure is similar to that of laboratory rabbits with an end piece thicker than that of human sperm. Morphometry data could be used for construction of a male gamete data base for further studies. PMID- 15551751 TI - Arrangement of beta tubulin in bull spermatozoa during nuclei decondensation. AB - Eukaryotic cells have internal scaffolding of microtubules cytoskeleton that gives them their characteristic shapes. We analyzed by immuno-fluorescence the shift and distribution of tubuline during in vitro bull sperm nuclei swelling by the action of heparin-reduced glutathione physiological decondensing agents. Sperm tubulin display a pattern that shows tubulin fluorescence all over the head, leaving the acrosome tip devoid of tubulin. In the second stage we can observe that the acrosomal zone is practically devoid of fluorescence and a net of fluorescent microtubules that seems to be anchored in the basal plate in the postacrosomal region. It is also possible to observe green spots of tubulin fluorescence in the nucleus periphery, that might represent clusters of chromatin hub-like bodies and/or the microtubule organizing center (MTOC). In the third stage, practically all tubulin moves backwards to the basal plate in the neck region of the sperm nuclei remaining in only the green fluorescence spots in the periphery of the swollen sperm nuclei. The results allow us to assume that tubulin mechanism rearrangement is considered to be necessary for the normal fertilization process. PMID- 15551752 TI - Less NO production and better motion parameter in human sperm by swim-up processing. AB - Fifteen semen specimens were obtained from men for semen analysis; each was divided into two aliquots for prepararation. The motile sperm recovery rate, percentage motility, and motion parameters were measured for each semen specimen (n = 15) before and after preparation with the use of the two methods, and cultured with different time courses (1 hr, 3 hr, and 6 hr). Nitric oxide (NO) was measured using the chemiluminscence method after centrifugation. Recovery rate of motile cell was significantly higher in direct swim-up method (69.5 +/- 42.4% versus 49.3 +/- 29.3%, p < 0.05). In motility, direct swim-up method in the different time courses was significantly better than IxaPrep method. (1 hr: 91.1 +/- 5.2% vs 65.6 +/- 16.4%, 3 hr: 87.2 +/- 7.9% vs 65.2 +/- 16.5%, 6 hr: 86.1 +/- 7.5% vs 60.8 +/- 17.6% and prewash: 61.6 +/- 16.2%, p < 0.05). In VAP and VSL, the sperm prepared by the above two methods all improved compared to pre-wash sperm (p < 0.05), but there was no statistical significance between the two methods. NO production in the direct swim-up group was significantly lower than IxaPrep group in the first hour of culture (0.09 +/- 0.09 uM vs 0.15 +/- 0.09 uM, p < 0.05). NO production increased as the culture time increased in swim-up group, but conversed in IxaPrep group. The lower level of NO produced in the swim up group may suggest that better sperm quality achieved is due to the decreased NO production. PMID- 15551753 TI - Interferon alpha-2b may impair testicular histology including spermatogenesis in a rat model. AB - Interferon-alpha has been used in various diseases at the reproductive ages. However, the effect of interferon-alpha on testicular histology has not been studied in literature. The aim of this study was to investigate the effects of interferon alpha-2B on testicular histology including spermatogenesis in a rat model. Seventeen adult male Wistar albino rats were divided into 3 groups: Six rats received 7.500 units (5 MIU/m2) of interferon alpha-2B (Intron), considered clinical treatment dose range. Six rats received 30.000 units (20 MIU/m2) of interferon alpha-2B (Intron), considered high treatment dose. Five rats served as a control group receiving 0.5 mL of saline injection. All injections were done intraperitoneally 3 times weekly for 3 weeks under inhalation anesthesia. All rats underwent bilateral orchiectomy 30 days after the experiment. Histological examination included the mean seminiferous tubular diameter (STD), germinal epithelial cell thickness (GECT), and testicular biopsy score (TBS). The mean STD was significantly lower in the low-dose interferon and high-dose interferon groups than in the control group (p = 0.008 and p = 0.004, respectively). The mean GECT was significantly lower in the low-dose interferon and high-dose interferon groups than in the control group (p = 0.008 and p = 0.004, respectively). The mean TBS was significantly lower in the low-dose interferon group (p = 0.05) and the high-dose interferon group (p = 0.01) than in the control group. The decreases in the mean values of the STD, GECT and TBS were not related to the interferon dose. Interferon alpha-2B may impair testicular histology even in clinical widely used treatment dose. Therefore, men at the reproductive ages should be fully informed for the use of interferon-alpha in the treatment of various diseases. PMID- 15551754 TI - A spectrum of clinical manifestations caused by host immune responses against Epstein-Barr virus infections. AB - Epstein-Barr virus (EBV), or human herpesvirus 4 (HHV-4), infects the vast majority of adults worldwide, and establishes both nonproductive (latent) and productive (lytic) infections. Host immune responses directed against both the lytic and latent cycle-associated EBV antigens induce a diversity of clinical symptoms in patients with chronic active EBV infections who usually contain an oligoclonal pool of EBV-infected lymphocyte subsets in their blood. Episomal EBV genes in the latent infection utilize an array of evasion strategies from host immune responses: the minimized expression of EBV antigens targeted by host cytotoxic T lymphocytes (CTLs), the down-regulation of cell adhesion molecule expression, and the release of virokines to inhibit the host CTLs. The oncogenic role of latent EBV infection is not yet fully understood, but latent membrane proteins (LMPs) expressed during the latency cycle have essential biological properties leading to cellular gene expression and immortalization, and EBV encoded gene products such as viral interleukin-10 (vIL-10) and bcl-2 homologue function to survive the EBV-infected cells. The subsequent oncogenic DNA damage may lead to the development of neoplasms. EBV-associated NK/T cell lymphoproliferative disorders are prevalent in Asia, but quite rare in Western countries. The genetic immunological background, therefore, is closely linked to the development of EBV-associated neoplasms. PMID- 15551755 TI - 2-methoxyestradiol enhances p53 protein transduction therapy-associated inhibition of the proliferation of oral cancer cells through the suppression of NFkappaB activity. AB - Protein transduction therapy using poly-arginine peptide can deliver the biologically active proteins. A previous study showed that 11 poly-arginine fused p53 protein (11R-p53) effectively penetrated across the plasma membrane and inhibited the proliferation of oral cancer cells. However, the intracellular half life of the delivered protein was less than 36 h. Previous studies also showed that 2-methoxyestradiol (2-ME), an endogenous non-toxic estrogenic metabolite, induces the stabilization of the wild-type p53 protein in human cancer cells posttranscriptionally. In the present study, we examined whether 2-ME induced the stabilization of 11R-p53 and had an inhibitory effect on the proliferation of oral cancer cells. The application of 2-ME significantly enhanced the inhibitory effect of 11R-p53 on the proliferation of oral cancer cells. However, 2-ME had no effect on the intracellular half-life of 11R-p53 in oral cancer cells. Of interest is the finding that 2-ME suppressed the transcriptional activity of NFkappaB, which has an important role in tumorigenesis, but did not affect p53 transcriptional activity. These results suggest that 2-ME synergistically enhances the 11R-p53-induced inhibition of the proliferation of oral cancer cells through the suppression of NFkB transcription. PMID- 15551756 TI - Relationship between postural balance and knee and toe muscle power in young women. AB - Muscle power in the lower extremities and body sway were measured in 57 healthy young women volunteers in their 20's. Body sway was measured with a stabilimeter for 30 sec during two-leg standing, and for 10 sec during one-leg standing with the eyes open or closed, alternating between right and left legs (5 times each). The measured parameters of body sway were locus length per time unit, locus length per environmental area, environmental area, rectangle area, root mean square area, and the ratio of sway with eyes closed to sway with eyes open. Knee flexor and extensor power and toe flexor and abductor power were the measures representing lower extremity muscle power. The increase in sway with the eyes closed was more marked during one-leg standing than two-leg standing, as expected. We found that 36 of 57 subjects (62%) were unable to maintain one-leg standing with their eyes closed, and this failure correlated with marked body sway (P = 0.0086). Many subjects had one leg that was classified as stable and the other leg classified as unstable. Clearly, testing of both legs alternately with eyes closed is necessary to measure the full range of sway in subjects. Lower extremity muscle power did not appear to be the dominant factor in maintaining balance in these young subjects. PMID- 15551757 TI - Expression and intracellular localization of FKHRL1 in mammary gland neoplasms. AB - FKHRL1 (FOXO3a), a member of the Forkhead family of genes, has been considered to be involved in the development of breast tumors; however, the in vivo expression and activation status of FKHRL1 in breast tumors still remains unclear. We immunohistochemically demonstrated the expression and intracellular localization of FKHRL1 in human breast tumors by the novel anti-FKHRL1 antibody which is available for formalin-fixed paraffin-embedded specimens. In a total of 51 cases of benign tumors, FKHRL1 was diffusely expressed in all cases, and its intracellular localization was revealed to be cytoplasmic (inactive form) in 94% of cases of intraductal papillomas (16/17) and 91% cases of fibroadenomas (31/34), with a similar pattern to normal glandular epithelium. In invasive ductal carcinomas, 83% of the cases (93/112) diffusely expressed FKHRL1; however, unlike benign tumors, 71% of the cases (66/93) showed the nuclear-targeted, active form of FKHRL1. Moreover, activated FKHRL1 was predominantly observed in scirrhous (29/36, 81% of the cases) and papillotubular (30/38, 79% of the cases) subtypes, compared to the solid-tubular subtype (7/19, 37% of the cases). Furthermore, the cases with nuclear-targeted FKHRL1 showed a tendency to have lymph nodal metastasis with statistical significance (P < 0.0001). Thus, the activation of FKHRL1 seems to be recognized as one of the specific features of invasive ductal carcinoma of the breast. PMID- 15551758 TI - Biofilm formation among methicillin-resistant Staphylococcus aureus isolates from patients with urinary tract infection. AB - Staphylococci have been confirmed to form biofilms on various biomaterials. The purpose of this study was to investigate biofilm formation among methicillin resistant Staphylococcus aureus (MRSA) isolates from patients with urinary tract infection (UTI) and to assess the relationship between biofilm-forming capacities and virulence determinants/clinical background. Over a 12-year period from 1990 through 2001, a total of 109 MRSA isolates were collected from patients (one isolate per patient) with UTI at the urology ward of Okayama University Hospital. We used the in vitro microtiter plate assay to quantify biofilm formation. We then investigated the presence of several virulence determinants by polymerase chain reaction assay and found eight determinants (tst, sec, hla, hlb, fnbA, clfA, icaA, and agrII) to be predominant among these isolates. Enhanced biofilm formation was confirmed in hla-, hlb-, and fnbA-positive MRSA isolates, both individually and in combination. Upon review of the associated medical records, we concluded that the biofilm-forming capacities of MRSA isolates from catheter related cases were significantly greater than those from catheter-unrelated cases. The percentage of hla-, hlb-, and fnbA-positive isolates was higher among MRSA isolates from catheter-related cases than those from catheter-unrelated cases. Our studies suggest that MRSA colonization and infection of the urinary tract may be promoted by hla, hlb, and fnbA gene products. PMID- 15551759 TI - Ureteroscopy using a detachable access sheath. AB - Ureteroscopy has evolved in many aspects, particularly in the flexibility and size of ureteroscopes. We have developed a new detachable access sheath to make ureteroscopic procedures more straight-forward and to reduce possible damage to delicate instruments used in the procedure. PMID- 15551760 TI - Metastatic carcinoma of the colon similar to Crohn's disease: a case report. AB - A 68-year-old Japanese man with a history of linitis plastica carcinoma of the stomach and subsequent gastrectomy 8 years previously presented with lower abdominal pain. Radiological and endoscopic examinations showed multiple submucosal nodular lesions similar to Crohn's disease in the ileocecal area. A firm diagnosis could not be made after initial multiple biopsies. Finally, a submucosal biopsy revealed adenocarcinoma. The ileocecal lesion was diagnosed as a recurrence because of the histological findings, which included mucosal preservation, a similarity with the histologic type of stomach carcinoma, and atypical immunoreactivity for primary colon carcinoma; the lesion was negative for both cytokeratin 7 and cytokeratin 20. In cases where metastatic carcinoma of the colon is suspected, we recommend early consideration of a submucosal biopsy. PMID- 15551761 TI - [Virtual colonoscopy in the screening of colon cancer]. PMID- 15551762 TI - [Physical exercise as a treatment of chronic neck pain]. PMID- 15551763 TI - [Technology and logistics of health care]. PMID- 15551764 TI - [Health of children born after in vitro fertilization]. PMID- 15551765 TI - [Vestibular neuronitis, sudden onset of peripheral vestibular disorder]. PMID- 15551767 TI - [Acute bronchitis]. PMID- 15551766 TI - [Exercise of neck and shoulder muscles as a relief for the chronic neck pain]. PMID- 15551768 TI - [Strange anomalies of the eye]. PMID- 15551769 TI - [Eye to eye]. PMID- 15551770 TI - [From gold to tissue engineering, direction towards the optimal correction and replacement]. PMID- 15551771 TI - [Artificial skin equivalents]. PMID- 15551772 TI - [Bioactive glass in the surgery of head and neck]. PMID- 15551773 TI - [New materials revolutionizing odontology]. PMID- 15551774 TI - [Biodegradable materials as tools to fasten bone in face and jaw surgery]. PMID- 15551775 TI - [Drug implants, technology serving delivery]. PMID- 15551776 TI - [Ophthalmological applications for biomaterials]. PMID- 15551777 TI - [Materials for artificial joints]. PMID- 15551778 TI - Transportation tribulations. PMID- 15551779 TI - The bell tolls for LNT. AB - The linear no-threshold (LNT) model has been a convenient tool in the practice of radiation protection, but it is not supported by scientific data at doses less than about 100 millisievert or at chronic dose rates up to at least 200 millisievert per year. Radiation protection practices based on the LNT model yield no demonstrable benefits to health when applied at lower annual doses. The assumption that such exposures are harmful may not even be conservative and has helped to foster an unwarranted fear of low-level radiation. For its new recommendations, to be issued probably in 2005, the International Commission on Radiological Protection (ICRP) has said that it expects to continue the application of the LNT model "above a few millisievert per year." National societies for radiation protection may wish to consider the need to lobby the ICRP, through the auspices of International Radiation Protection Association, to further relax adherence to the LNT assumption--up to "a few tens of millisievert per year." PMID- 15551780 TI - An evaluation of the Kearny Fallout Meter (KFM), a radiation detector constructed from commonly available household materials. AB - A radiation detector constructed of common household materials was developed at Oak Ridge National Laboratory (ORNL) by Cresson H. Kearny and has been referred to as the Kearny Fallout Meter (KFM). Developed during the height of the Cold War, the KFM was intended to place a radiation meter capable of measuring fallout from nuclear weapons in the hands of every U.S. citizen. Instructions for the construction of the meter, as well as information about radiation health effects, were developed in the form of multi-page newspaper insert. Subsequently, the sensitivity of the meter was refined by a high school teacher, Dr. Paul S. Lombardi, for use in demonstrations about radiation. The meter is currently being marketed for survivalists in light of potential radiation terrorist concerns. The KFM and Lombardi's variation of it are constructed and evaluated for this work. Calibrated tests of the response and variations in response are reported. A critique of the multi-page manual is made. In addition, the suitability of using such a detector, in terms of actual ease of construction and practical sensitivity, is discussed for its use in demonstrations and introductory classes on nuclear topics. PMID- 15551782 TI - A NATO exercise on radiological sampling. AB - The North Atlantic Treaty Organization (NATO) has held its first-ever large-scale radiological exercise employing unsealed radioactive sources. The objective of the exercise was to validate NATO protocols on radiological sampling and surveying. However, the exercise also proved to be a valuable training opportunity and was highly instructive to all involved. This paper highlights the lessons learned from this exercise, particularly in the areas of radiation survey equipment and techniques, sampling techniques, and field measurements. PMID- 15551781 TI - A database management system for radiation safety program activities. AB - The Database Management System includes a data retrieval system for most aspects of the radiation safety program at our institution. Originally, this program was designed to simplify the retrieval of data related to radioisotope use at the institution. After several modifications and changes, the present system is more elaborate and uses current commercial software, which is more readily available to the entire university and hospital community. The data included in the database are radioactive material receipts and inventory; radioisotope waste inventory and disposal; quarterly laboratory inspection results; contamination (wipe tests) results and instrument inventory and calibration; and support staff training. Both the authorized users and the Radiation Safety Office personnel maintain records independently using the same format, which makes for easy comparison and verification. Data for and from laboratory inspections are readily accessed, analyzed, and reported. The system allows for modifications as necessitated by changes in local and federal regulations or in-house policies. PMID- 15551783 TI - In situ gamma spectroscopy measurement of 41Ar during neutron activation analysis with the SLOWPOKE II reactor in Jamaica. AB - In situ gamma spectroscopy was used to measure 41Ar released into the laboratory due to the activation of air in the irradiation tubes of SLOWPOKE reactor during routine neutron activation analysis. The data obtained were used to predict the 41Ar distribution in the laboratory for various operating conditions of the reactor. The dose received by the analyst from the immersion in 41Ar was calculated to be 2.36 microSv gamma(-1), which is approximately 1% of the normal background exposure. PMID- 15551784 TI - Internal dosimetry--a review. AB - The field history and current status of internal dosimetry is reviewed in this article. Elements of the field that are reviewed include standards and models, derivation of dose coefficients and intake retention fractions, bioassay measurements, and intake and dose calculations. In addition, guidance is developed and provided as to the necessity of internal dosimetry for a particular facility or operation and methodology for implementing a program. A discussion of the purposes of internal dosimetry is included as well as recommendations for future development and direction. PMID- 15551785 TI - Light water reactor health physics. AB - In this article an overview of the historical development of light water reactor health physics programs is presented. Operational health physics programs have developed and matured as experience in operating and maintaining light water reactors has been gained. Initial programs grew quickly in both size and complexity with the number and size of nuclear units under construction and in operation. Operational health physics programs evolved to face various challenges confronted by the nuclear industry, increasing the effectiveness of radiological safety measures. Industry improvements in radiological safety performance have resulted in significant decreases in annual collective exposures from a high value of 790 person-rem in 1980 to 117 person-rem per reactor in 2002. Though significant gains have been made, the continued viability of the nuclear power industry is confronted with an aging workforce, as well as the challenges posed by deregulation and the need to maintain operational excellence. PMID- 15551786 TI - Latency and the lung cancer epidemic among United States uranium miners. AB - The latency of occupational cancer was a key factor in the recent epidemic of lung cancer among U.S. uranium miners. A review of the epidemic and analysis of latency periods with a near lifetime follow-up found that among former and nonsmokers, the mean mid-induction latent period is nearly a constant at about 25 y, regardless of age at starting or magnitude of exposure. Among cigarette smokers, the mean is shorter (about 19 y). It is not influenced by age at start of smoking, amount smoked, or magnitude of exposure, but there is a marked shortening as the age at start of radiation exposure rises. These latency variables affect lifetime risk models. By disregarding the European radon mine exposures and waiting for strong evidence of lung cancer among U.S. uranium miners (ignoring the exposures occurring while waiting during the latency period), the epidemic became inevitable. PMID- 15551787 TI - A modified ICRP 66 iodine gas uptake model and its parametric uncertainty. AB - Intakes via inhalation may occur from radionuclides released in the form of a gas. The chemical characteristics pertaining to the release influence the intake and subsequent dose to an exposed individual. Gases are taken up or absorbed in the entire respiratory tract and the associated uptake mechanisms are quite different from deposition of particulates. Gaseous iodine can exist in various chemical forms, e.g., elemental iodine, inorganic, and organic iodine compounds. These different chemical species play an integral role in the gaseous uptake o f iodine in t he respiratory tract. Gas uptake in the various regions of the respiratory tract results in the intake of iodinated material into the body. The radioactive iodine taken up in the gas-exchange tissues is absorbed into the bloodstream of an individual and subsequently transferred to other organs. Iodine in the circulatory system can then be taken up by the thyroid gland, with resulting dose to the thyroid. The magnitude and uncertainty in regional gas uptake is important in the assessment of individuals exposed to airborne releases of radioiodine. The current ICRP 66 model is rudimentary and estimates regional gas uptake based on solubility and reactivity of the different radionuclides entering the respiratory tract. The modified model proposed here employs methodology and a mathematical structure to determine estimates of fractional gas uptake rather than defaulting to literature values, as in the current ICRP model. Model parameters have been assigned input distributions and estimates of uncertainty have been determined. A sensitivity analysis of these parameters has been performed to demonstrate the importance of each of these parameters. The sensitivity analysis ranks the model-input parameters by their importance to estimates of regional gas uptake. The model developed herein may be used for improved estimation of gas uptake in the respiratory tract and subsequent dose estimates from the different chemical forms of radioiodine. PMID- 15551788 TI - Specific absorbed fractions for internal photon emitters calculated for a tomographic model of a pregnant woman. AB - Specific absorbed fractions are essential for calculation of radiation dose from internal emitters. Existing specific absorbed fractions for pregnant women were calculated using the stylized models; in this work, a partial-body tomographic model for a pregnant woman was constructed from a rare set of CT images. Based on this tomographic model, the Monte Carlo code, EGS4-VLSI, was used to derive specific absorbed fractions. Monoenergetic, isotropic photon emitters from 15 keV to 4 MeV were distributed in different source organs, and doses were calculated to many target regions in the body. Even though the results showed general agreement with previous studies for higher energies, significant differences were also found, especially for lower energies. The main reasons for the differences are due to the variation of mass, geometry, and organ distances, and they demonstrate the influence of more realistic body models on dose calculations. PMID- 15551789 TI - The deposition and translocation of methyl iodide by crops. AB - Organic forms of radioactive iodine are released during routine and accidental releases from the nuclear industry. Methyl iodide is often the predominant species in these releases. This paper describes the results of a study to determine the deposition, allocation, and loss of radioiodine after crops of bean, carrot, and cabbage were exposed to CH3 125I. The deposition velocity ranged from 0.14-7.10 x 10(-4) cm s(-1), which is in line with previous studies. Translocation of radioiodine away from the leaves to other crop components was observed post exposure. The partition of radioiodine was to those crop components growing most actively at the time of exposure. This finding contradicts some previous studies and will have implications for dose assessments. Losses of radioiodine were only observed as a consequence of leaf fall. The consequences of these findings for the modeling the movement of radioiodine in crops following the deposition of methyl iodide are discussed. PMID- 15551790 TI - Resrad-recycle: a computer model for analyzing radiation exposures resulting from recycling radioactively contaminated scrap metals or reusing radioactively surface-contaminated materials and equipment. AB - RESRAD-RECYCLE is a computer code designed by Argonne National Laboratory (ANL) to be used in making decisions about the disposition of radioactively contaminated materials and scrap metals. It implements a pathway analysis methodology to evaluate potential radiation exposures resulting from the recycling of contaminated scrap metals and the reuse of surface-contaminated materials and equipment. For modeling purposes, it divides the entire metal recycling process into six steps: (1) scrap delivery, (2) scrap melting, (3) ingot delivery, (4) product fabrication, (5) product distribution, and (6) use of finished product. RESRAD-RECYCLE considers the reuse of surface-contaminated materials in their original forms. It contains representative exposure scenarios for each recycling step and the reuse process; users can also specify scenarios if desired. The model calculates individual and collective population doses for workers involved in the recycling process and for the public using the finished products. The results are then used to derive clearance levels for the contaminated materials on the basis of input dose restrictions. The model accounts for radiological decay and ingrowth, dilution and partitioning during melting, and distribution of refined metal in the various finished products, as well as the varying densities and geometries of the radiation sources during the recycling process. A complete material balance in terms of mass and radioactivity during the recycling process can also be implemented. In an international validation study, the radiation doses calculated by RESRAD-RECYCLE were shown to agree fairly well with actual measurement data. PMID- 15551791 TI - Ecological study of solar radiation and cancer mortality in Japan. AB - Geographic observation of the increased mortality of some cancers at higher latitudes has led to a hypothesis that vitamin D produced after exposure to solar radiation has anti-carcinogenic effects. However, it is unclear whether such association would be observed in countries like Japan, where fish consumption, and therefore dietary vitamin D intake, is high. Pearson correlation coefficients were calculated between averaged annual solar radiation levels for the period from 1961 through 1990 and cancer mortality in the year 2000 in 47 prefectures in Japan, with adjustments for regional per capita income and dietary factors. A moderate, inverse correlation with solar radiation was observed for cancers of the esophagus, stomach, colon, rectum, pancreas, and gallbladder and bile ducts in both sexes (correlation coefficient, ranging from -0.6 to -0.3). The results of this study support the hypothesis that increased exposure to solar radiation reduces the risk of cancers of the digestive organs. PMID- 15551793 TI - Objective nursing notes: are they a possibility? PMID- 15551792 TI - Measurements of occupational exposure for a technologist performing 18F FDG PET scans. AB - Radiation doses to one PET technologist performing 100 18F FDG (18F fluorodeoxyglucose) imaging procedures were measured in a clinical setting using two types of thermoluminescent dosimeter (TLD) badges, one finger-ring TLD and one electronic pocket dosimeter (EPD). 18F FDG was handled either with unshielded or with viewing window tungsten shielded syringes. The resulting doses using unshielded syringes were 13.8 +/- 0.8 microSv/370 MBq and 14.3 +/- 0.4 microSv/370 MBq, measured with TLD 100 and with TLD 700H/600H, respectively. For the same series of measurements, the doses obtained using shielded syringes were 10.7 +/- 0.4 microSv/370 MBq and 7.2 +/- 2.1 microSv/370 MBq with TLD700H/600H and with EPD, respectively. The dose to the right hand from shielded syringes was 69.3 +/- 5.5 microSv/370 MBq. All these values are within the ICRP recommended dose limits. Extrapolated to 725 examinations per year, the resulting effective dose measured with TLD would be 10 mSv with unshielded and 7.5 mSv with shielded syringes, respectively (25% dose reduction). The doses measured by TLD were consistently higher than those measured by EPD, suggesting that EPD measurements might underestimate occupational doses. PMID- 15551794 TI - Using the stages of change model to improve communication about advance care planning. AB - TOPIC: Advance care planning (ACP) is a term intended to describe the dynamic process of discussing wishes for future medical care. PURPOSE: To demonstrate how a behavior change model can be used to enhance ACP. SOURCES: Authors' personal experience, workshops, literature review. CONCLUSIONS: Incorporating behavioral change theory helps develop workable interventions and provides strategies for nurses to approach patients appropriately about planning for future medical care. PMID- 15551795 TI - Threat: a concept analysis for a new era. AB - TOPIC: Living in a post-September 11 world, the concept of threat has never been more apparent in people's minds. PURPOSE: To reviews past analyses, capture the critical elements, and clarify a newly defined meaning of threat. SOURCES: Walker and Avant's method was used to conduct the analysis. CONCLUSIONS: Understanding an individual's perceived threat and the emotions experienced by this phenomenon can help develop research-based interventions, decrease perceived threat, increase positive coping strategies, and decrease negative emotions. PMID- 15551796 TI - The importance of prayer for mind/body healing. AB - TOPIC: A growing interest in using complementary and alternative modalities (CAM) to treat the patient as a whole. PURPOSE: To address the use of prayer for mind/body healing in nursing practice. SOURCES: Published research articles, studies, book chapters. CONCLUSIONS: Key research studies show that CAM are gaining acceptance by healthcare providers. Prayer is especially growing as an alternative method for healing. Additional research is needed to demonstrate further the effectiveness of prayer as a CAM modality for mind/body healing. PMID- 15551797 TI - A retrospective look at the entry dilemma. AB - Nursing is multidimensional, interactive, interdisciplinary, and complex. Almost anything that can be said about nursing can be said another way. Some things worth being said and heard will not follow the norms of journal presentation. A forum accommodates the emerging voice, the new format, the innovative approach. Nursing Forum, in an effort to honor the independent voice in nursing, presents here the voice who elects to enter the dialogue, but who does so "in another way." PMID- 15551799 TI - The role of magnetic resonance imaging in the diagnosis of postoperative spondylodiscitis. AB - BACKGROUND: Spondylodiscitis, discitis associated with vertebral osteomyelitis may follow disc-removal surgery. A targeted successful treatment of spinal infections requires clinical and laboratory data that are completed by the contribution of imaging procedures. Neuroimaging provides precise information on correct topography, localization, propagation, and differential diagnosis of spinal infectious lesions. The aim of this study was to present magnetic resonance imaging (MRI) findings in patients with postoperative spondylodiscitis. METHODS: MRI was performed in 6 patients aged 29-50, with clinically suspected postoperative spondylodiscitis. Initial examination was performed 3-8 weeks after surgery and 3, 6, or 12 months after the treatment by antibiotics. Patients underwent MRI on a IT imaging unit (Siemens, Magnetom-Impact), including sagittal T1W and T2W images and axial T1W images before and after the administration of gadolinium contrast medium. RESULTS: MRI findings included: significantly decreased signal intensity with the loss of distinction between vertebral body and intervertebral disc space on T1W, increased signal intensity in the adjacent vertebral body and end-plates on T2W, contrast enhancement of vertebral body and disc space and paravertebral soft tissue changes. Follow-up examinations performed 3, 6, or 12 months after the treatment showed less abnormal signal intensities on both T1- and T2-weighted images. CONCLUSION: Postoperative spondylodiscitis is a rare but severe complication of lumbar disc surgery. Since conventional imaging techniques are not reliable for detecting spondylodiscitis in its early stages, MRI is of great significance in the diagnosis of postoperative spondylodiscitis. PMID- 15551798 TI - Comparison of two different protocols for the induction of maturation of human dendritic cells in vitro. AB - BACKGROUND: Dendritic cells (DC) have been used for immunotherapy of malignant tumors, different kinds of infections, and other clinical conditions. For that purpose, optimal conditions for the generation of functionally mature DC in vitro are required. Two different protocols for the induction of maturation of monocyte derived DC (MDDC) were compared in this study. METHODS: MDDC were generated in vitro by cultivating adherent monocytes of healthy volunteers with granulocyte macrophage-colony stimulating factor (GM-CSF) and interleukin 4 (IL-4) during 6 days period. The immature DC thus prepared were induced to mature using two protocols. DC were stimulated for 2 days with lipopolysaccharide (LPS), or with a cocktail of proinflammatory mediators (PM) containing IL-1beta, IL-6, tumor necrosis factor alpha (TNFalpha), and prostaglandin E2 (PGE2), respectively. Phenotypic characteristics of MDDC and their endocytic activity were studied by flow cytometry. Allostimulatory activity of these cells was tested in the mixed leukocyte reaction (MLR), whereas the production of cytokines was determined by ELISA kits. RESULTS: MDDC matured with PM (PM-DC) were predominantly non-adherent cells, while about 30% of LPS-matured DC were adherent cells. In comparison with LPS-DC, PM-DC expressed higher levels of CD86 and CD83, had lower endocytic activity, produced higher levels of IL-10 and lower levels of IL-12, and more strongly stimulated proliferation of allogeneic lymphocytes. CONCLUSION: The protocol based on the combination of proinflammatory cytokines and PGE2 is better for the induction of maturation of human MDDC in vitro than the protocol using LPS alone. PMID- 15551800 TI - Antiepileptic, behavioral, and antidepressant effects of adjuvant lamotrigine therapy in drug-resistant epilepsy. AB - AIM: To evaluate the behavioral effects of lamotrigine as add-on therapy in treatment-resistant epilepsy. METHODS: An open, prospective, long-term study of lamotrigine as adjuvant therapy was performed in 56 patients with drug-resistant epilepsy (female/male ratio 35/21, age range 16-51 years). All the patients kept seizure diaries, and electroencephalograms were recorded at baseline and during 24 months of the treatment. Quality of life questionnaire, Hamilton depression scale (HMD), Beck depression scale (BDI), and Hamilton anxiety scale (HMA) were used before and during lamotrigine therapy. Comparative assessments were made in an age- and sex-matched control group treated with other antiepileptic drugs. RESULTS: Overall, seizure control was improved in 55.3% of the patients, remained unchanged in 39.3%, and deteriorated in 5.4%. Improvement in some quality of life measures occurred in 50% of the patients. The HMD subscales and BDI scale showed significant improvement in lamotrigine treated patients compared to the control group (ANOVA, p < 0.01). Negative behavioral effects occurred in 10.7% of the patients. CONCLUSION: Lamotrigine demonstrated significant antiepileptic long term efficacy, and its positive effects on the mood and quality of life, which surpassed the negative behavioral effects, and contributed highly to the favorable treatment outcome. PMID- 15551801 TI - [Endothelial trauma in the surgery of cataract]. AB - Cataract surgery is most common in human surgery and comprises 80% of eye surgery programs. Owing to sophisticated technologies, it has become a routine surgery with lowered complications rate; hence, the functional outcome is more conditioned by operative trauma. The aim of this study was to demonstrate the significance of specular microscopy in the evaluation of operative trauma during extracapsular cataract extraction (ECCE) and phacoemulsification (P), in a controlled environment. The study included 100 consecutive patients who met the established criteria, and groups were formed according to the type of surgery by the assignment of successive numbers from a random number table. Examination and photographs of the corneal endothelium, as well as pachymetry were performed on Keeler-Konan Poclington Specular Microscope (KSP). The obtained results revealed significant dissimilarity in endothelial cell reduction (9.17% in group E, and 4.72% in group P), which generated statistically significant correlation of preoperative and post-operative pachymetry in the group E (p=0.0004). On the basis of the results obtained by specular microscopy, it was concluded that under the same conditions phacoemulsification caused reduced operative trauma of the corneal endothelium. PMID- 15551802 TI - [Morphological changes in aorto-coronary vein graft--the analysis of autopsy and biopsy material]. AB - BACKGROUND: Patients with implanted aortic coronary grafts have different survival time, which raises the question why the efficacy of graft implants is so poor. The aim of this study was to present the results of the analysis of morphological changes in the vein grafts taken after the death of patients who died after surgery in different time intervals, as well to present the analysis of the grafts obtained after surgical reintervention. METHODS: The total number of 656 grafts of 308 dead patients was analyzed, as well as 76 grafts from 40 patients who underwent surgical reintervention. According to the duration of the graft since surgical intervention until death, all the analyzed changes were divided into two groups: a) early changes and complications, and b) late changes and complications in aorto-coronary vein grafts. RESULTS: After the autopsy, 518 vein grafts from the first group were evaluated histopathologically. Changes were found in the form of small or large areas with peeled endothelium in 266 grafts, with the insudation of fibrin and thrombocytes in such places, subendothelial edema, and occlusive thrombosis of the graft lumen. Significant stenosis, which occurred distally from the anastomoses, was present in 118 grafts without changes in the walls of the graft, and there was significant narrowing of the graft lumen in 134 vein grafts due to intimal hyperplasia. In the second group, 138 grafts were histopathologically analyzed after autopsy. Significant hyperplasia was present in 117 grafts with the migration of smooth muscle cells from media into intima, and in 21 grafts there were atheromatous plaques. In 120 veins analyzed before the graft implantation, the lesion or the lack of endothelium was found, as well as the penetration of fibrin and blood elements and intimal hyperplasia. In 46 veins analyzed before the graft implantation, significant intimal hyperplasia with the elevated number of smooth muscle cells was found. CONCLUSION: The most frequent lesions in the grafts were the lesions of the endothelium, which caused thrombosis formation and lumen occlusion. Intimal hyperplasia in patients with longer survival time occurred due to the migration of smooth muscle cells from the media, or due to the formation of atherosclerotic plaques, which caused graft lumen stenosis or thrombosis. PMID- 15551803 TI - [Evaluation of local and regional spread of malignant tumors of the tongue and floor of the mouth]. AB - Malignant tumors of the oral cavity grow rapidly, frequently and early metastazing to the surrounding regional lymph nodes. The aim of this study was to evaluate the correlation between clinically confirmed local and regional spread and intraoperatively and histopathologically verified local spread. A series of 74 patients with carcinoma of the tongue and floor of the mouth were analyzed. All the patients were surgically treated during the period 1991-1995. Clinical evidence of local spread (cT) was in high accord with intraoperatively and histopathologically evidenced spread (pT) amounting to 83.8%. The degree of correlation decreased with the increase of T stage. Clinically observed regional spread (cN) and intraoperatively and histopathologically confirmed regional spread (pN) was lower, amounting to 56.8% in comparison to the corresponding T categories. PMID- 15551804 TI - Neuroendocrine differentiation in prostate cancer. AB - BACKGROUND: In numerous recent studies attention has been focused to neuroendocrine differentiation (NED) in prostate cancer (PC). Focal NED is present in almost all PCs, but it is prominent in only 5-10% of the carcinomas. The prognostic significance of focal NED in PC is controversial, but current evidence suggests its influence on the onset and/or conversion of hormone resistant tumor phenotype. The aim of this study was to evaluate the relationship between NED status, based only on immunohistochemical use of neuroendocrine (NE) markers, with PC grade and stage, and preoperative serum levels of prostate specific antigen (PSA). METHODS: The study included the biopsy material of 73 untreated PC patients (pts.) obtained by transurethral resection (TUR) (37 pts.), and radical retropubic prostatectomy (RRP) (36 pts.). Two representative tissue samples (typically the block containing the largest amount of neoplasm) were selected for immunohistochemical (IMM) staining. NE cells were identified using a panel of IMM markers: chromogranin A, neuron-specific enolase, and serotonin. The level of PC exocrine differentiation was detected by monoclonal antibodies against PSA. RESULTS: Significant expression of NE cells was demonstrated in 26 (70.2%) pts. with PC after TUR. In this group, serum preoperative PSA values ranged from 0.1 to 9.6 ng/ml. The majority of pts. with NED had low differentiated PC with Gleason grade score (GGS) > 7, and normal PSA values below 4 ng/ml (77%), in clinical stage D (54%). Statistically significant correlation (p<0.01) of positive NED with higher stage and grade and low PSA values was established. Among the pts. with localized PC in whom RRP was performed (n=36), significant expression of NE cells was found in 15 pts. (41.7%), 8 (53.3%) in pT2 stage, and 7 (46.7%) in pT3 stage. Significant correlation between NED with preoperative PSA values and stage of PC in pts. with RRP was not found. CONCLUSION: We demonstrated the significant NED in poorly differentiated PC in patients in the advanced stage of the disease. The expression of NED in organ confined PC did not correlate with tumor stage, but it correlated with tumor grade (GGS < or = 7). PMID- 15551805 TI - Autologous bone marrow-derived progenitor cell transplantation for myocardial regeneration after acute infarction. AB - BACKGROUND: Experimental and first clinical studies suggest that the transplantation of bone marrow derived, or circulating blood progenitor cells, may beneficially affect postinfarction remodelling processes after acute myocardial infarction. AIM: This pilot trial reports investigation of safety and feasibility of autologous bone marrow-derived progenitor cell therapy for faster regeneration of the myocardium after infarction. METHODS AND RESULTS: Four male patients (age range 47-68 years) with the first extensive anterior, ST elevation, acute myocardial infarction (AMI), were treated by primary angioplasty. Bone marrow mononuclear cells were administered by intracoronary infusion 3-5 days after the infarction. Bone marrow was harvested by multiple aspirations from posterior cristae iliacae under general anesthesia, and under aseptic conditions. After that, cells were filtered through stainless steel mesh, centrifuged and resuspended in serum-free culture medium, and 3 hours later infused through the catheter into the infarct-related artery in 8 equal boluses of 20 ml. Myocardial viability in the infarcted area was confirmed by dobutamine stress echocardiography testing and single-photon emission computed tomography (SPECT) 10-14 days after infarction. One patient had early stent thrombosis immediately before cell transplantation, and was treated successfully with second angioplasty. Single average ECG revealed one positive finding at discharge, and 24-hour Holter ECG showed only isolated ventricular ectopic beats during the follow-up period. Early findings in two patients showed significant improvement of left ventricular systolic function 3 months after the infarction. There were no major cardiac events after the transplantation during further follow-up period (30-120 days after infarction). Control SPECT for the detection of ischemia showed significant improvement in myocardial perfusion in two patients 4 months after the infarction. Echocardiographic assessment in these two patients also showed significant improvement of systolic function three months after the infarction. CONCLUSION: Preliminary results of the study showed that the transplantation of bone marrow-derived progenitor cells into the infarcted area was safe, and feasible, and might improve myocardial function. Further follow-up will show if this treatment is effective in preventing negative remodeling of the left ventricle and reveal potential late adverse events (arrhythmogenicity and propensity for restenosis). PMID- 15551806 TI - [Botulinum toxin A in the treatment of anal fissure]. AB - BACKGROUND: This paper presents our first experience in the treatment of primary anal fissure by injection of botulinum A toxin into the internal sphincter. METHODS: The study group of the retrospective study included 12 outpatients (8 females and 4 males), mean age 42 (range 18-60). During the period 2000-2003, after unsuccessful conservative treatment, patients were treated with the injections of botulinum A toxin, 100 units on both sides of the anal fissure laterally into the internal anal sphincter (50 units on either side). The patients were clinically evaluated 3, 7, and 30 days, and 3 and 6 months after the treatment. RESULTS: Three fissures had healed after a month, and seven after 3 months. Two remained unhealed, but asymptomatic. There was no incontinence of flatus or faeces after 3 months of the treatment. After temporary healing, two fissures relapsed after 6 months, and these patients had the adequate tonus of the anal sphincter muscles. Except for the temporary incontinence, there were neither other side-effects, nor serious complications. CONCLUSION: Injection of botulinum A toxin achieved good results in the treatment of anal fissure. The appropriate use makes this method safe as an alternative to surgical treatment of anal fissure. PMID- 15551807 TI - Renewed granulocyte support practice and its alternatives. PMID- 15551808 TI - [War injuries of the extremities]. PMID- 15551809 TI - Psoriasis in a patient with dermatomyositis. AB - Psoriasis has been consistently associated with arthritis and inflammatory bowel diseases, though there have been reports on patients with psoriasis and other autoimmune dermatoses. Sometimes, sharply demarcated scaly plaques located over extensor surfaces in patients with dermatomyositis may clinically resemble psoriatic lesions. Histologic findings of interface dermatitis, typical for dermatomyositis, help clinicians to rule out psoriasis. A patient is presented with dermatomyositis in which psoriatic lesions developed over the extremities and lower trunk. Histological examination confirmed the diagnosis of psoriasis. Both diseases have run independent courses. It is prudent to include psoriasis in the differential diagnosis of scaly eruptions occurring in dermatomyositis patients. PMID- 15551811 TI - A well-differentiated liposarcoma co-existent with leiomyoma. AB - BACKGROUND: The coexistence of a well-differentiated liposarcoma (atypical lipomatous tumor) and benign smooth muscle component in a single soft tissue neoplasm is extremely rare. CASE REPORT: Histologic and immunohistochemical characteristics of tumor, which represented the recurrence of a retroperitoneal atypical lipoma in a 50-year-old female patient are presented in this paper. The tumor represented the recurrence of a retroperitoneal atypical lipoma. Lipomatous component consisted of a mixture of lipoma-like and sclerosing variant of well differentiated liposarcoma. Characteristically, the heterologous smooth muscle differentiation was manifested as a macroscopically visible nodular growth in the form of a leiomyoma within the adipocytic component. It consisted of intersecting fascicles of spindle cells, which lacked mitoses and significant atypia, and were immunopositive for smooth muscle markers. CONCLUSION: This case of well differentiated liposarcoma with benign smooth muscle is the first reported tumor of retroperitoneal localization. It is also the first one to exhibit the heterologous smooth muscle component as a distinct leiomyoma. PMID- 15551810 TI - [Umbilical metastasis (Sister Mary Joseph's nodule) diagnosed by fine-needle aspiration]. AB - Sister Mary Joseph's nodule is the eponym for metastatic involvement of the umbilicus. This less common entity is the sign of disseminated malignant disease, mainly of digestive and gynecologic origin, and is associated with a poor prognosis. A case of Sister Mary Joseph's nodule in a 76-year-old woman in whom the umbilical metastasis was the first sign of malignant disease in presented. The diagnosis of metastatic adenocarcinoma was established by fine needle aspiration cytology of the umbilical nodule. Radiological and ultrasonographic investigation disclosed carcinoma of the gallbladder with pancreas, stomach, and colon invasion as well as peritoneal dissemination. The diagnosis was confirmed by exploratory laparatomy and histological examination of the excised umbilical nodule. PMID- 15551813 TI - [Colors for the patient rooms of the 3E Hospital. Community lecture to students of the course 2001/2004]. PMID- 15551814 TI - [Myoma: many women have unnecessary surgery -- finally new methods come to Germany, too]. PMID- 15551812 TI - Smallpox outbreak in Yugoslavia in 1972. PMID- 15551815 TI - [Grief attendance in child's death approximately at birth]. PMID- 15551816 TI - [Measles-mumps-rubella vaccine and autism -- rumors become soon not to be extinguished]. PMID- 15551817 TI - [Changes in vocational image of the hospital nurse]. PMID- 15551818 TI - [Prebiotic ballast stuff. New developments in infant nutrition]. PMID- 15551819 TI - [Not performing the duties in occupational activity can have far-reaching legal liability outcome]. PMID- 15551820 TI - [Health care reform leaves allergy and skin patients in the lurch]. PMID- 15551822 TI - [National Commission on Breast Feeding (BfR). Recommendations on duration of breast feeding (discharged on March 1, 2004)]. PMID- 15551821 TI - [When the deodorant gives up -- help against excessive sweating]. PMID- 15551823 TI - Biosorption of heavy metals by red algae (Palmaria palmata). AB - The biosorption of heavy metals from aqueous solutions was investigated, using a cheap and abundant dry biomass of red algae P. palmata. The Freundlich, Langmuir and Brunauer Emmer and Teller (BET) models were used to describe the uptake of lead (pb2+), copper (Cu2+), nickel (Ni2+), cadmium (Cd 2+) and zinc (Zn2+) on P. palmata. The good fits of the Langmuir and BET models to the experimental data reflected that the sorption on P. palmata was a multi-layer sorption, in which a Langmuir equation could be applied to each layer. The highest maximum sorption capacity q(max), derived from the Langmuir model was 15.17 mg g(-1) for lead and 6.65 mg g(-1) for copper (dry weight metal/dry weight biosorbent) at a pH of 5.5 6. The affinity of metals for P. palmata was found to decrease in the order: Pb2+ > Cd2+ > Cu2+ > Ni2+. The factors influencing copper and lead uptake were found to be contact time, pH, initial concentration and temperature. Biosorption of copper and lead was a rapid process, with 70% and 100% of the respective uptakes occurring within the first 10 minutes. PMID- 15551824 TI - Startup of an anaerobic fluidized bed reactor for landfill leachate treatment. AB - Anaerobic treatment of the young landfill leachate in a pilot-scale fluidized bed reactor was performed to observe the effects of startup on COD removal, biogas production and biomass attachment. During the experiments, the organic loading rate (OLR) was increased gradually from 2.5 to 27 g COD l(-1) day(-1) with varying the feed rate and the hydraulic retention time in the range of 5.5-13 1 day(-1) and 1-2.35 days, respectively. The anaerobic fluidized bed reactor (AFBR) showed an excellent performance and attained steady state conditions with COD removal of 90% after 80 days. Biogas production in the reactor continuously increased and its average yield (Y(gas)) was defined as 0.531 biogas g(-1) COD(rem)(-1) with a correlation of 99%. Greater microbial attachment was found at the increased loading rates and the attached biomass concentration (X(attach)) consisted of about 90% of the total biomass concentration. In addition, the substrate utilization rate indicated a linear increase as a function of time during the process. PMID- 15551825 TI - Phosphate/sulphate exchange studies on Amberlite IRA-400. AB - The sorption behaviour of phosphate on strong basic anion exchanger, Amberlite IRA-400 (SO4(2-) form) is studied as a function of pH (3-11) at two different initial concentration ranges (0.161 - 0.807 mmol l(-1), 3.874 - 6.134 mmol l(-1)) at 25 degrees C. The stiochiometry of the exchange reactions is observed to be dependent upon concentration and pH of the solutions. It is suggested that the sorption of phosphate takes place initially as PO4(3-) followed by HPO4(2 )/H2PO4(-). The data obtained is explained with the help of a modified Langmuir and mass law action equations. PMID- 15551826 TI - Treatment of dairy wastewater using a vertical bed with passive aeration. AB - The aim of this research was to investigate the feasibility of treating liquid dairy wastes by a vertical bed equipped with an innovative passive aeration system. The vertical bed (32 liter) was operated by recirculating consecutive batches of liquid waste in the column. Batches of liquid waste were applied at two different rates: 1) each batch was recirculated for 72 hours, and 2) each batch was recirculated for 24 hours. Settled liquid dairy wastes (5000 mg l(-1) COD, 2000 mg l(-1) BOD and 2500 mg l(-1) TSS) were used in the experiments. When the reactor operated with each batch recirculating for 72 hours, the BOD and COD reduction were 66% and 40%, respectively. The vertical bed operated successfully without the need for an additional rest period. The main removal was observed to take place during the first 20 hours. No biomass or solids accumulation was observed indicating that the remaining 52 hours of recirculation were actually used for bed regeneration, i.e. integrated rest period. When the reactor operated with each batch recirculating for 24 hours, the system clogged after 21 days. An additional 24 day rest period was needed in order to free 94% of the initial void space. In this mode, the BOD and COD reduction were 67% and 47%, respectively. The overall COD removal in a complete operational cycle (feeding period followed by a rest period) was 467 g COD m(-3) d(-1) (996 g COD m(-2) d(-1)). This value is 1.4 higher than the COD removal obtained in the 72 hour per batch mode and shows the advantage of conventional vertical bed operation of intensive feeding followed by rest period rather than a rest period integrated into the feeding cycle. PMID- 15551827 TI - Catalytic oxidation of SCN- and CN- ions from aqueous solutions. AB - Oxidation of inorganic contaminants, such as thiocyanate and cyanide ions, by ozone in water has improved significantly in the presence of solid catalysts based on nickel and cobalt oxides supported on alumina. It can be considered that the hydroxyl radical produced by catalytic decomposition of ozone over the catalyst's surface was responsible for the oxidation of the adsorbed ions. Ozonation efficiency was increased by 20% in the presence of catalytic systems in particular for those systems containing both nickel and cobalt oxide. PMID- 15551828 TI - A new technology for automatic identification and sorting of plastics for recycling. AB - A new technology for automatic sorting of plastics, based upon optical identification of fluorescence signatures of dyes, incorporated in such materials in trace concentrations prior to product manufacturing, is described. Three commercial tracers were selected primarily on the basis of their good absorbency in the 310-370 nm spectral band and their identifiable narrow-band fluorescence signatures in the visible band of the spectrum when present in binary combinations. This absorption band was selected because of the availability of strong emission lines in this band from a commercial Hg-arc lamp and high fluorescence quantum yields of the tracers at this excitation wavelength band. The plastics chosen for tracing and identification are HDPE, LDPE, PP, EVA, PVC and PET and the tracers were compatible and chemically non-reactive with the host matrices and did not affect the transparency of the plastics. The design of a monochromatic and collimated excitation source, the sensor system are described and their performances in identifying and sorting plastics doped with tracers at a few parts per million concentration levels are evaluated. In an industrial sorting system, the sensor was able to sort 300 mm long plastic bottles at a conveyor belt speed of 3.5 m.sec(-1) with a sorting purity of -95%. The limitation was imposed due to mechanical singulation irregularities at high speed and the limited processing speed of the computer used. PMID- 15551829 TI - Sulfur formation by steady-state continuous cultures of a sulfoxidizing consortium and Thiobacillus thioparus ATCC 23645. AB - The elemental sulfur formation by the partial oxidation of thiosulfate by both a sulfoxidizing consortium and by Thiobacillus thioparus ATCC 23645 was studied under aerobic conditions in chemostat. Steady state was attained with essentially total conversion to sulfate when the dissolved oxygen concentration was 5 mgO2 l( 1) and below a dilution rate (D) of 3.0 d(-1)for the consortium and 0.9 d(-1) for T thioparus. The consortium formed elemental sulfur in steady state under oxygen limitation. Fifty percent of the theoretical elemental sulfur yield was obtained with a dissolved oxygen concentration of 0.2 mgO2 l(-1). Growth of T thioparus was negatively affected with a concentration below 1.9 mgO2 l(-1). Consortium yield from batch cultures was 2.1 g(-1) (protein) mol(-1) (thiosulfate), which was comparable with the values obtained in the chemostat at dilution rates of 0.4 d(-1) and 1.2 d(-1). The consortium showed a maximum degradation rate of 0.105 g(thiosulfate) g(-1) (protein) min(-1) and a saturation rate for S2O3(2-) of 1.9 mM. PMID- 15551830 TI - Solubilization and desorption of PAHs in soil-aqueous system by biosurfactants produced from Pseudomonas aeruginosa P-CG3 under thermophilic condition. AB - Surface-active agents (surfactants) can enhance bioremediation of soils contaminated with PAHs by reducing sorption of PAHs or increasing desorption rates. The effectiveness of nonionic surfactant (Tween 80) and biosurfactants to enhance the solubilization and desorption of phenanthrene (Phe) and pyrene (Pyr) in soil-aqueous systems under thermophilic conditions was investigated using batch studies. Tween 80 and biosurfactants produced from Pseudomonas aeruginosa strain, ATCC 9027 and a strain isolated in our laboratory, P-CG3 were tested in this study. The results showed that all the surfactants could effectively enhance the solubility of both Phe and Pyr under thermophilic condition (55 degrees C), which was linearly proportional to the concentrations of surfactants at concentrations above their respective critical micelle concentration (CMC). When the surfactants' concentrations in aqueous phase were above their respective CMCs, substantial amounts of PAHs were desorbed from soil into the aqueous phase. Among the three surfactants used, the biosurfactant produced from our own isolate, P-CG3 was more effective in enhancing the solubilization and desorption of PAHs, implying that it might have the potential to be further applied in the bioremediation of PAH contaminated soils. PMID- 15551831 TI - Biodegradation of Red B dye by Bacillus sp. OY1-2. AB - Batch tests were employed to investigate the effects of aerobic and anoxic conditions on the biodegradation of Red B dye by Bacillus OY1-2. Results from batch experiments demonstrated anoxic conditions were beneficial for rapid biodegradation of Red B dye in comparison to aerobic conditions. Biodegradation is a major mechanism in the decolorization of Red B dye in comparison with biosorption, which accounted for only 8% of the total decolorization efficiency. Reactors packed with granular activated carbon (GAC) and inoculated with Bacillus OY1-2 were investigated to treat a synthetic wastewater under anoxic conditions. In the absence of cosubstrates, Red B dye was degraded; however, a significant improvement in degradation resulted with the addition of cosubstrates. PMID- 15551832 TI - Physical and chemical processes for removing suspended solids and phosphorus from liquid swine manure. AB - A physical/chemical treatment train, that included 24-hour preliminary settling followed by coagulation/flocculation and sedimentation, was tested at a laboratory bench scale to treat liquid swine manure for the removal of total suspended solids (TSS) and total phosphorus (TP). Preliminary (i.e., natural) settling time had an effect on TSS removal within only the first 24 hours. TSS removal efficiency reached 75% (TSS concentration was reduced from 5,800 to 1,450 mg 1(-1)) after 24 hours of preliminary settling. Also, as a result of the 24 hour preliminary settling, TP concentration was reduced from 533 to 318 mg 1(-1), thus leading to a TP removal efficiency of 40%. When compared to ferric chloride, alum was more effective in reducing both TSS and TP. At a 95% confidence interval, alum dose, coagulation Gt (coagulation velocity gradient * rapid-mixing time), and flocculation Gt (flocculation velocity gradient * slow-mixing time) were not significant for TSS removal while alum dose was the only significant factor for TP removal. For the 24-hour settled liquid manure that had a TP concentration in the range of 362 to 401 mg l(-1)and as alum dose increased up to 1,600 mg 1(-1), TP removal efficiency increased up to 70%. Then, the rate of increase in TP removal efficiency per increase in alum dose started to decrease and TP removal efficiency reached about 93% at an alum dose of 3,000 mg 1(-1). Sequential alum dosing improved the TSS removal efficiency while it had no effect on TP removal efficiency. The mass ratio of removed TSS/applied alum increased from about 0.38, during a one-step dosing of alum at a concentration of 1,600 mg l(-1), to about 0.58 during a two-step dosing of alum at a concentration of 1,600 mg l(-1) (i.e., 800 mg l(-1) followed by another 800 mg l(-1)). PMID- 15551834 TI - Nitrate removal from saline water using autotrophic denitrification by the bacterium Thiobacillus denitrificans MP-1. AB - Autotrophic denitrification of synthetic wastewater by Thiobacillus denitrificans MP-1 isolated from mangrove sediment was investigated in both up-flow packed-bed reactors and fermentor. More than 97.5% and 90% of the nitrate in inflow was removed after 8.8 and 161 hours at 250 and 195 mg l(-1) for the packed-bed reactor and fermentor system, respectively. The nitrate was quickly denitrified at very low flow rates (0.11 m h(-1)) for the packed-bed reactors, but as the flow rate was greater than 0.13 m h(-1), the nitrate removal rate increased as the flow rate increased. In the static fermentor system, the denitrification can be described by a secondary reaction, but at a flow rate between 1.31 to 1.49 m h(-1), the reactor performance can be described using the zero-order reaction in the packed-bed reactor. As the speed increases, the zero-order reaction translates into half-order reaction as the penetration efficiency of nitrate decreases. The mass ratios between the nitrate removed and the sulfate produced were determined to be 6.81 and 9.32 in the reactor column and fermentor, respectively. The results of this study suggest that efficient removal of high concentrations of nitrate in water or wastewater can be achieved effectively using autotrophic bacteria immobilized on surfaces of sulphur granules in packed bed reactor. PMID- 15551833 TI - Performance of a UASB-digester system treating domestic wastewater. AB - The anaerobic treatment of raw domestic wastewater by a novel technology consisting of an Up-flow Anaerobic Sludge Bed (UASB) reactor combined with a completely mixed digester for the stabilisation of the UASB sludge was assessed. A pilot-scale plant of the so-called UASB-Digester system was located at the municipal wastewater treatment facility of Santiago de Compostela (Northwest of Spain). The main aim of the Digester was to enhance the biodegradation of influent solids retained in the UASB reactor at low temperatures, then increasing its specific methanogenic activity. The sludge drawn from the middle zone of the UASB entered the upper zone of the Digester and then circulated from the bottom of the Digester to the UASB bottom. Circulating in an automated semi-continuous way, the flow of this sludge stream was selected in order to set a previously defined hydraulic retention time (HRT) (16-27 d) in the digester. The Digester temperature was set at an optimum value ranging from 25 to 35 degrees C. The steady state efficiency of the UASB system, at 6-8 h of HRT, 15-16 degrees C of temperature and 330-360 mg l(-1) of influent total chemical oxygen demand (TCOD) was 79% of total suspended solids (TSS) removal, 52% of TCOD removal and 60% of biological oxygen demand (BOD5) removal. The hydrolysis of retained solids reached 85%, while excess sludge generation was only 7% of influent TCOD. A stable anaerobic (pre)treatment of diluted domestic wastewater was reached as the sludge concentration in the reactor remained mainly constant and the specific methanogenic activity showed a slight increase. PMID- 15551835 TI - Adolescent accidents and injury: prevention is the way. PMID- 15551836 TI - Youth injury in Israel: an appeal for change. AB - Unintentional injury in children and adolescents have raised public health concerns in recent years, because it is the leading cause of death, a physical burden for the injured person, an emotional and social burden for the families together with an economic burden for society. The National Center for Children's Safety and Health in Israel was established in 1995 at the Schneider Children's Medical Center in order to focus on prevention of childhood and adolescent injury and safety. Over these years data has been collected from the Central Bureau of Statistics, the Ministry of Health, the National Trauma Register, Police and the National Poison Center in order to look for trends and develop strategies for intervention. These efforts, together with the active prevention programs that have been established, resulted in a national report (1) presented to the Minister of Health and other policy makers in 2003. The current situation of youth injuries in Israel, which over the years have not shown any improvement, necessitates a new agenda with a focus on prevention and collaboration between various ministries and agencies in order to implement it. The report alone cannot have an impact upon youth safety, but we hope that it will provide an impetus for developing youth safety strategies and programs based on data. We also believe there is a need for long-term monitoring and follow-up of youth injury trends in Israel. PMID- 15551837 TI - Burn injuries and adolescents in Israel. AB - Burn injury is a public health concern often associated with individual pain, emotional stress, prolonged hospitalizations, permanent disfigurement and family stress. In this paper we studied the avaliable data on burn injury among adolescents in Israel through a Medline search and found three relevant studies with data on this population. The incidence rate of burn injury was 0.46 per 1,000 children aged 5-14 years for Jews and 0.91 for Bedouin. Most of the burn injury in this age group was caused by hot liquids, followed by fire and chemical burns for both Jews and Bedouin, but electical burns occurred more often in Bedouins. Mortality was very low for the adolescent group. Prevention programs in schools since the 1980s have been found effective, but the public health focus should now be geared towards groups at risk. PMID- 15551838 TI - Adolescent injury risk behavior. AB - Health-risk behaviors that contribute to the leading causes of morbidity and mortality among adolescents and young adults are often established during adolescence and extended into adulthood. Unintentional motor vehicle injury is the leading cause of mortality in childhood and adolescence in developed countries. This review presents some of the risk factors found in research on unintentional injury and death in adolescence, including risk factors for siblings and adolescents with intellectual disability. These findings should be connected with the findings of a recent study that showed that about one third of all unintentional childhood injury deaths in the United States were preventable. For injury prevention to take place and being effective a multidisciplinary approach is needed to identify host, agent and environmental factors using epidemiology research and biomechanics. In the population of adolescents with intellectual disability there has been little research on injury epidemiology or injury prevention and the service provider will need to focus and educate staff on this issue in order to prevent injury that can result in further disability. PMID- 15551839 TI - Neighborhood social composition and injury risks among pre-adolescent and adolescent boys and girls. A study in Stockholm metropolitan. AB - The study investigates the extent to which social and socioeconomic characteristics of a population within a particular living area influence injury risks among young people. The study group comprised pre-adolescent and adolescent boys and girls aged 10-19 living in the Stockholm metropolitan area in Sweden over the three-year period 2000--2002 (about 185,000 subjects each year). Area comparisons were made at parish level (96 parishes) based on three compositional indexes derived from a factor analysis of sixteen population attributes. Thereafter, each factor was transformed into an additive index and divided into three levels. Diagnosis-specific injury risks were then measured by index, considering injury causes with documented social differences (five for boys and three for girls). Injuries resulting in at least one night of hospitalization during the period 2000--2002 were considered. Three main dimensions with regard to the social fabric of the Stockholm metropolitan area were identified: socioeconomic precariousness and ethnic concentration (Factor 1), educational and financial assets (Factor 2), and concentration of well-off citizens of Nordic origin (Factor 3). Lower levels of socioeconomic precariousness and ethnic concentration showed a protective effect on boys in the cases of traffic and sports-related injuries, but an aggravating one in the cases of falls on the same level and violence-related injuries. Level of educational and financial assets did not impact on falls on the same level among boys, but increased the risk of such injuries among girls. Increased risks of traffic-related injuries among boys and of falls on the same level among both boys and girls were found in areas with lower concentrations of well-off citizens of Nordic origin. It is concluded that social and socioeconomic composition of the population in a living area impacts on injury risks of various kinds in a rather specific manner--in magnitude and in kind. The mechanisms via which contextual aspects operate during youth are likely to vary according to type (cause) of injury and gender. PMID- 15551840 TI - Alcohol and road safety behaviour among New Zealand tertiary students. AB - In New Zealand, as in most developed countries, rates of injury mortality and morbidity peak in late adolescence (15-19 years), when they account for around three quarters of fatalities and a third of hospitalisations. Road traffic crashes account for over half of fatal injuries in New Zealand as in most developed countries. Individuals engaged in post-secondary education, i.e., tertiary students, are a large and important subgroup of young people exposed to the risk of road traffic injury. The aims of this study were to estimate the prevalence of unsafe road behaviours and to examine the role of demographic variables, alcohol use, driver licence status, and transport needs. Participants were 1,480 students (899 women) from 12 residential halls in Dunedin, New Zealand. They anonymously completed a questionnaire on alcohol use and road safety at the start of the 2000 academic year. A sub-sample of 967 students completed a follow-up survey six months later. The mean age of the sample was 18.3 years (SD=1.6), and 88% had a driver licence. Male gender and drinking status were independently associated with speeding, drink-driving and drink riding, but not seatbelt use. Drinking levels showed a strong dose-response relationship with the prevalence of unsafe behaviours with the exception of seatbelt use. In marked contrast to drinking levels, which increased from baseline to follow-up, drink-driving and drink-riding levels decreased, probably reflecting the reduced motor vehicle transport needs of students in residential halls, relative to their out-of-term living environments. In addition to various existing, evidence-based countermeasures, application of interventions for reducing hazardous drinking may be required to help reduce road traffic injury rates in this population. PMID- 15551841 TI - Substance use and youth violence. A study among 6th to 10th grade Israeli school children. AB - This study examined the co-morbidity of substance use and violence among a representative sample of 8,394 6th-10th grade Israeli students. A representative national self report sample of 8,394 students in 6th through 10th grade. Measures included smoking, alcohol consumption, and illicit drug use, predicting involvement in bullying, injury during a fight and weapon-carrying in the past 30 days. We found across all grades, genders and ethnicities, daily smoking, use of hard drugs, history of drunkenness and binge drinking were the best predictors of violent behavior. Involvement in such behaviors put girls in higher risk for violent behaviors compared with boys. We concluded that use of substances immensely increased the odds of involvement in violent behavior, and this association was extremely strong for Arab girls. The study suggested that although girls were less frequently involved in substance use, the girls who did were at much higher risk for involvement in youth violence. PMID- 15551842 TI - Is there equalisation in socioeconomic differences in the risk of traffic injuries in childhood? A study of three cohorts of Swedish school children. AB - Equalisation of socioeconomic differences in injury risks at school ages is investigated, considering changes in risk inequalities over time for various categories of road users. Three national cohorts of children were followed up after their entry in different compulsory school levels (about 190,000 subjects by cohort). Subjects were attributed a household socioeconomic status in 1990 and their traffic injuries were sought over a five-year period (1990--94) in the national hospital discharge and causes of death registers. Socioeconomic inequality was measured by year and cohort for various RTI (Road Traffic Injury) diagnoses using the Relative Index of Inequality (RII). Whether equalisation arose or not was assessed considering changes in RII scores within the five-year period, measured by Chi-squared test. RII scores were generally high except for bicycle-related injuries among boys from the oldest cohort throughout the follow up. This relative equality in bicycle-injury risk benefited boys from all socioeconomic groups. Also, there was some, though short lasting, equalisation in bicycle-related injury risk for the younger cohort at the end of the first grade of compulsory school. Hence, some but limited support to the concept of equalisation was found, especially among young male bicycle riders. Population based intervention aimed at improving safety behaviours among bicycle or motor vehicle users should target not only suspected class-related influences, but even influences from the school, peer group and youth culture. Transitional periods in modes of transportation are a major concern. PMID- 15551843 TI - Unintentional injuries among 16 to 20 year old students in Switzerland. AB - In our society, accidents constitute a major public health problem, especially among youth. The objective of this paper was to describe the incidence of nonfatal injuries that required medical care among 16 to 20 year-old in Switzerland, its distribution by type of injury and whether there were differences by gender or by academic track and whether these injuries had sequels (hospitalisation, physical and psychological sequels). Overall, 28.3% of the sample reported at least one accident needing medical care in the previous 12 months, with males having more accidents than females and apprentices more than students. By type of accident, sports were the most frequently reported, followed by traffic, leisure time and work accidents. Half of males and one-third of females reported more than one accident, and 16% and 8% of them, respectively, reported four or more. Both physical and psychological sequels were more frequent among females, while hospitalisation was more frequent among males. Accident prevalence rates remain high among adolescents. Safety counselling and environmental measures need to be implemented. PMID- 15551844 TI - Adolescents, accidents and the need to extend our scope of research. AB - Accidents in adolescence result in mortality and disability that in many cases could have been prevented. The issue is a multi-factorial complex phenomenon, where new research ideas are needed to improve outcome. This short communication presents several research ideas that could provide new relevant answers. PMID- 15551845 TI - Trends in adolescent injury mortality in Israel. AB - Epidemiology is an important and powerful tool in public health used to monitor health, observe trends and identify risk factors for utilization in strategy, policy and planning of services and interventions. Unintentional injury is the leading cause of death in adolescents worldwide with motor vehicle injury and drowning the leading causes. In Israel the mortality rate for motor vehicle mortality rate for adolescents was found to be 10.6 per 100,000 and 1.0 per 100,000 for drowning. Studies have shown a decrease in injury and mortality over time, but there is still much to do and calculation studies have shown that it is possible to prevent further with targeted serious effort and strategies employed. PMID- 15551846 TI - The importance of safety and quality in rural America. PMID- 15551847 TI - The environmental context of patient safety and medical errors. AB - The environmental context of patient safety and medical errors was explored with specific interest in rural settings. Special attention was paid to unique features of rural: healthcare organizations and their environment that relate to the patient safety issue and medical errors (including the distribution of patients, types of adverse events associated with learning, information flows, triage and transfer decisions, and culture of safety). Relevant organizational theories and strategies fo medical error reduction and prevention in rural health care settings were identified. Financial and technical assistance are needed to support the systematic collection of data from rural hospitals and other entities and to enhance relevant patient safety practices for rural America. PMID- 15551848 TI - Assuring rural hospital patient safety: what should be the priorities? AB - CONTEXT: Since reports on patient safety were issued by the Institute of Medicine, a number of interventions have been recommended and standards designed to improve hospital patient safety, including the Leapfrog, evidence-based safety standards. These standards are based on research conducted largely in urban hospitals, and it may not be possible to generalize them to rural hospitals. PURPOSE: The absence of rural-relevant patient safety standards and interventions may diminish purchaser and public perceptions of rural hospitals, further undermining the financial stability of rural hospitals. This study sought to assess the current evidence concerning rural hospital patient safety and to identify a set of rural-relevant patient safety interventions that the majority of small rural hospitals could readily implement and that rural hospitals, purchasers, consumers, and others would find relevant and useful. These interventions should help rural hospitals prioritize patient safety efforts. METHODS: As background, we reviewed literature; interviewed representatives of provider, payer, consumer, and governmental groups in 8 states; and calculated patient safety indicator rates in rural hospitals using the Agency for Healthcare Research and Quality's Health Care Cost and Utilization Project National Inpatient Sample. Based on the research literature and patient safety recommendations from national organizations, we developed a list of potentially important patient safety areas for rural hospitals. The rural relevance of these safety interventions was evaluated by a national expert panel in terms of the frequency of the problem, ability to implement, and the internal and external value to rural providers, purchasers, and consumers. FINDINGS: The limited available research suggests that patient safety events and medical errors may be less likely to occur in rural than in urban hospitals. We identified 9 areas of patient safety and 26 priority patient safety interventions relevant to rural hospitals. CONCLUSIONS: Many of the identified areas of patient safety and interventions are relevant to all types of hospitals, not just rural hospitals. However, some areas, such as transfers, are especially relevant to rural hospitals. The challenges of implementing some interventions, such as 24/7 pharmacy coverage, are significant given workforce supply and financial problems faced by many small rural hospitals. The results of this study provide an important platform for further work to test the validity and effectiveness of these interventions. PMID- 15551849 TI - Quality improvement strategies and best practices in critical access hospitals. AB - CONTEXT: Critical access hospitals (CAHs) face many challenges in implementing quality improvement (QI) initiatives, which include limited resources, low volume of patients, small staffs, and inadequate information technology. A primary goal of the Medicare Rural Hospital Flexibility Program is to improve the quality of care provided by CAHs. PURPOSE: This article describes key quality improvement initiatives for a national sample of CAHs that are actively involved in implementing quality-related initiatives in collaboration with support hospitals and statewide organizations. METHODS: Researchers conducted a national telephone survey of 72 CAHs and 2 in-depth case studies of CAHs. FINDINGS: The survey and case studies demonstrate that many CAHs are successfully implementing QI activities, including patient safety initiatives, improvements in overall QI processes and peer review processes, and implementation of QI projects focused on treatment of 1 or more specific diseases. The CAHs are involved with multiple external organizations in these activities. The administrators of the 2 case study CAHs have made QI a priority for their hospitals; ensured that resources are available for QI activities; and worked with their support hospitals, statewide organizations, and other CAHs to develop and implement rural-relevant QI initiatives. CONCLUSIONS: Cost-based Medicare reimbursement has been a key factor in the ability of CAHs to fund additional staff, staff training, and equipment to improve patient care. The commitment of hospital leaders and key staff is a crucial factor in moving QI initiatives forward in CAHs. PMID- 15551851 TI - What would be the effect of referral to high-volume hospitals in a largely rural state? AB - CONTEXT: Volume of certain surgical procedures has been linked to patient outcomes. The Leapfrog Group and others have recommended evidence-based referral using specific volume thresholds for nonemergent cases. The literature is limited on the effect of such referral on hospitals, especially in rural areas. PURPOSE: To examine the impact of evidence-based referral by volume standard for 5 hospital procedures (abdominal aortic artery repair, coronary angioplasty, coronary artery bypass graft, esophageal cancer surgery, and pancreatic resection) in a largely rural state. METHODS: Healthcare Cost and Utilization Project Iowa State Inpatient Dataset was analyzed to identify hospitals meeting the volume standard versus those not meeting the standard. FINDINGS: Relatively few hospitals perform these procedures in Iowa. Hospitals performing the procedures at a volume above the threshold standard set by the Leapfrog Group tend to be larger, receive more transfers from other hospitals for these procedures, and perform fewer of these procedures on an emergency basis. In Iowa, hospitals that met the volume standard did not differ from hospitals that did not meet the volume standard in risk-adjusted mortality rates. The impact of evidence based referral would be substantial in terms of travel time for some procedures (ie, coronary artery bypass graft, pancreatic resection, and esophageal cancer surgery) and produce considerable lost revenue for some hospitals. CONCLUSIONS: Evidence-based referral would be associated with substantial burden for some patients and hospitals in Iowa. This negative impact does not appear to be offset by improvement in in-hospital mortality rates. These initial findings suggest that there are a number of issues that need to be considered, especially in a rural state, before evidence-based referral is embraced as a means to enhance patient outcomes. PMID- 15551850 TI - Translating research into practice: voluntary reporting of medication errors in critical access hospitals. AB - CONTEXT: Low service volume, insufficient information technology, and limited human resources are barriers to learning about and correcting system failures in small rural hospitals. PURPOSE: This paper describes the implementation of and initial findings from a voluntary medication error reporting program developed by the Nebraska Center for Rural Health Research (NCRHR) to overcome these barriers in 6 Nebraska critical access hospitals (CAHs). METHODS: Participating Nebraska CAHs mailed copies of medication error reports to the NCRHR monthly for entry into a database. Quarterly summaries enabled each CAH to compare its reports by severity, type, phase of the medication use process, contributing factors, and causes to those of its peers and MEDMARX, a national medication error reporting program. Workshops emphasized learning from the reported errors by identifying system sources of variation in medication use and initiating change to achieve best practices. FINDINGS: Similar to MEDMARX, 99% of medication errors reported by 6 Nebraska CAHs were not harmful, reported errors most often originated in the administration phase, and the most common error type was omission. The CAHs reported significantly smaller proportions of "near miss" errors and errors originating in the prescribing phase than in MEDMARX. CONCLUSIONS: By collaborating with CAHs, an academic medical center, and a national reporting program, the NCRHR is translating the Institute of Medicine's recommendation for voluntary error reporting into practices that allow CAHs to learn about and improve their medication use systems. However, limited presence of pharmacists in CAHs is a barrier to implementing double checks and learning from system failures in the medication use system. PMID- 15551852 TI - Applied strategies for improving patient safety: a comprehensive process to improve care in rural and frontier communities. AB - CONTEXT: Medical errors and patient safety have gained increasing attention throughout all areas of medical care. Understanding patient safety in rural settings is crucial for improving care in rural communities. PURPOSE: To describe a system to decrease medical errors and improve care in rural and frontier primary care offices. METHODS: Applied Strategies for Improving Patient Safety (ASIPS) was a demonstration project designed to collect and analyze medical error reports and use these reports to develop and implement interventions aimed at decreasing errors. ASIPS participants were clinicians and staff in 2 practice based research networks: the Colorado Research Network (CaReNet) and the High Plains Research Network (HPRN). This paper describes ASIPS in HPRN. FINDINGS: Fourteen HPRN practices with a total of 150 clinicians and staff have participated in ASIPS. Participants have submitted 128 reports. Diagnostic tests were involved in 26% of events; medication errors appeared in 20% of events. Communication errors were reported in 72%. Two learning groups developed "Principles for Process Improvement" for medication errors and diagnostic testing errors. Several safety "alerts" were issued to improve care, and 2 interventions were implemented to decrease errors. CONCLUSIONS: A safe and secure reporting system that relies on voluntary reporting from clinicians and staff can be successfully implemented in rural primary care settings. Information from reports can be used to identify processes that can be improved. PMID- 15551853 TI - Organization of care for acute myocardial infarction in rural and urban hospitals in Kansas. AB - CONTEXT: One in 4 Americans lives in a rural community and relies on rural hospitals and medical systems for emergent care of acute myocardial infarctions (AMI). The infrastructure and organization of AMI care in rural and urban Kansas hospitals was examined. METHODS: Using a nominal group process, key elements within hospitals that might influence quality of AMI care were identified, including personnel, equipment, organizational systems, and quality improvement activities. These elements were included in a survey of 45 rural and 12 urban Kansas hospitals. FINDINGS: Though emergency 911 systems were widely available in both urban and rural communities, paramedics and advanced cardiac life support were less likely to be available in rural communities. Few rural hospitals were capable of emergent catheterization, angioplasty, or coronary artery bypass surgery; cardiologists, though readily available by phone, were rarely available on-site. Nevertheless, most rural ambulances could not bypass local hospitals. Most rural hospitals transferred the vast majority of their patients to urban medical centers within an average distance of 78 miles. Standardized protocols were used for emergent AMI care in 67% of urban and 62% of rural hospitals. Hospitals included aspirin in 53% and beta-blockers in 28% of either protocols or standing orders. CONCLUSIONS: Although faced with more limited resources, some rural hospitals, like their urban counterparts, have implemented protocols to address emergent care of AMI patients. Nevertheless, many of these protocols omit crucial aspects of AMI care. Rural and urban hospitals should jointly develop systems that assure consistent, rapid delivery of AMI care. PMID- 15551854 TI - Barriers to cancer screening by rural Appalachian primary care providers. AB - CONTEXT: Rural Appalachia has significantly higher overall cancer mortality compared with national rates, and lack of cancer screening is believed to be one of the contributing factors. Reducing the cancer disparity in this region must include strategies to address suboptimal cancer screening practices by rural Appalachian primary care providers (PCPs). PURPOSE: To identify and investigate barriers to recommending and/or performing cancer screening among rural Appalachian PCPs. METHODS: A semistructured focus group research design was used to elicit perceived barriers to recommending and/or performing cancer screening from 36 rural Appalachian PCPs (in 5 groups), including physicians, nurse practitioners, and a physician assistant. FINDINGS: Findings indicate that rural Appalachian PCPs may not be performing recommended cancer screenings for a number of reasons. Time constraints, conflicting guidelines, and perceptions that patients do not value prevention were reported barriers to cancer screening. The PCPs in this study expressed frustration in attempting to encourage cancer screening and cited patient factors such as socioeconomic status, Appalachian culture, and cancer fatalism as barriers to cancer screening. CONCLUSIONS: Rural Appalachian PCPs encounter various barriers, such as lack of time and multiple cancer screening guidelines, to incorporating cancer screening into their practice routine. The findings underscore the negative impact of some cultural factors on preventive care delivered by PCPs. Increased provider education is needed on how best to encourage cancer screening within a cultural context and should include clarification and understanding of current cancer screening guidelines. PMID- 15551855 TI - Comparative performance data for critical access hospitals. AB - CONTEXT: Among small rural hospitals, there is a growing recognition of the need to measure and report on the use of resources and the safety and quality of the services provided. Dashboards, clinical value compasses, and balanced scorecards are approaches to performance measurement that have been adopted by many health care organizations. However, there exists very little comparative performance data specific for critical access hospitals. PURPOSE: To identify how comparative performance data for critical access hospitals (CPD-CAH) might facilitate performance and quality improvement, to assess the potential benefits and drawbacks of such data, and to identify some of the critical issues in the development and implementation of CPD-CAH. METHODS: Assessment of discussions by participants at a rural hospital performance improvement summit and authors' analyses. FINDINGS: CPD-CAH potentially could improve quality of care and patient outcomes, provide comparative data and benchmarks, inform policy development, facilitate collaboration, and enhance community relations. However, CPD-CAH could also impose an unaffordable cost, produce poor information, require complex coordination, induce a negative public reaction, and result in perverse hospital behavior. Development and implementation of CPD-CAH would require including stakeholders' assessment of its desirability and feasibility, setting objectives, establishing guiding principles, developing a method, collecting and analyzing data, and disseminating results. CONCLUSIONS: CPD-CAH could significantly advance CAH performance and quality improvement. However, development and implementation would be a complicated exercise requiring academic expertise and practitioner consultation. The potential value of CPD-CAH should be carefully weighed against its potential cost. PMID- 15551856 TI - Measuring rural hospital quality. AB - CONTEXT: Increased interest in the measurement of hospital quality has been stimulated by accrediting bodies, purchaser coalitions, government agencies, and other entities. PURPOSE: This paper examines quality measurement for hospitals in rural settings. We seek to identify rural hospital quality measures that reflect quality in all hospitals and that are sensitive to the rural hospital context. METHODS: We develop a conceptual model for measuring rural hospital quality, with a focus on the special issues posed by the rural hospital context for quality measurement. With the assistance of a panel of rural hospital and hospital quality measurement experts, we review hospital quality measures from national and rural organizations for their fit to rural hospitals. FINDINGS: Based on this analysis, we recommend an initial core set of quality measures relevant for rural hospitals with less than 50 beds. This core set of 20 measures includes 11 core measures from the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) related to community acquired pneumonia, heart failure, and acute myocardial infarction; 1 measure related to infection control; 3 measures related to medication dispensing and teaching; 2 procedure-related measures; 1 financial measure; and 2 other measures related to the use of advance directives and emergency department monitoring of trauma vital signs. CONCLUSION: Based on the special measurement needs posed by the rural hospital context, we suggest avenues for future quality measure development for core rural hospital functions (eg, triage, stabilization, and transfer, and emergency care) not considered in existing quality measurement sets. PMID- 15551857 TI - Intensive care unit utilization and interhospital transfers as potential indicators of rural hospital quality. AB - CONTEXT: Obtaining meaningful information from statistically valid and reliable measures of the quality of care for disease-specific care provided in small rural hospitals is limited by small numbers of cases and different definitive care capacities. An alternative approach may be to aggregate and analyze patient services that reflect more generalized care processes. PURPOSE: To evaluate the applicability of intensive care unit (ICU) utilization and interhospital transfers as potential indicators of quality in rural hospitals. METHODS: Secondary data analysis of ICU utilization and interhospital transfer practices in Iowa's Critical Access (CAH), rural, rural referral, and urban hospitals. FINDINGS: Rural hospitals have fewer resources, provide a more limited range of definitive care services, and rely to a greater extent on transferring patients to other hospitals capable of providing the required definitive care. Examining ICU utilization and interhospital transfer patterns we found (1) that lower percentages of patients receive ICU care in smaller facilities; (2) higher transfer rates for both ICU and non-ICU patients in CAH hospitals; (3) shorter average lengths of stay for ICU patients from smaller hospitals who were transferred; and (4) lower mortality rates for CAH and rural hospital ICU patients. CONCLUSIONS: Examining ICU utilization and interhospital transfer patterns offers potential insights into rural hospital quality measurement and comparisons. PMID- 15551858 TI - Wolfgang Baumeister--Felix Hoppe-Seyler Lecturer 2004. PMID- 15551859 TI - Mapping molecular landscapes inside cells. AB - Cryoelectron tomography opens a window into the inner space of cells. It combines the potential of three-dimensional imaging with a close-to-life preservation of biological samples. Tomograms with molecular resolution are essentially images of the cellular proteome and, in conjunction with advanced pattern recognition techniques, they can be used to map the molecular landscape inside organelles and cells. PMID- 15551860 TI - Paper of the Year 2003: Award to Dieter Hoffmann. PMID- 15551861 TI - Structural basis of denitrification. AB - Denitrification represents an important part of the biogeochemical cycle of the essential element nitrogen. It constitutes the predominant pathway of the reductive dissimilation of nitrate in the environment. Via four enzymatic reactions, nitrate is transformed stepwise to nitrite (NO2-), nitric oxide (NO), and nitrous oxide (N2O), to finally yield dinitrogen gas (N2). All steps within this metabolic pathway are catalyzed by complex multi-site metalloenzymes with unique spectroscopic and structural features. In recent years, high-resolution crystal structures have become available for these enzymes with the exception of the structure for NO reductase. PMID- 15551862 TI - Roles of nectins in cell adhesion, migration and polarization. AB - Nectins are Ca2+-independent immunoglobulin (Ig)-like cell-cell adhesion molecules, which comprise a family consisting of four members. Nectins have five activities: (1) they show Ca2+-independent cell-cell adhesion activity by homo- and hetero-trans-interactions through their extracellular regions; (2) they bind afadin, an actin filament (F-actin)-binding protein, through their cytoplasmic tails and are connected to the actin cytoskeleton; (3) they induce activation of Cdc42 and Rac small G proteins through their cytoplasmic tails; (4) they bind Par 3, a cell polarity protein, through their cytoplasmic tails; and (5) they heterophilically trans-interact with Necls, nectin-like molecules, through their extracellular regions. Through these activities, nectins regulate a variety of cellular functions, including adhesion, migration, and polarization. Here we describe these activities and functions of nectins. PMID- 15551863 TI - Designing novel spectral classes of proteins with a tryptophan-expanded genetic code. AB - Fluorescence methods are now well-established and powerful tools to study biological macromolecules. The canonical amino acid tryptophan (Trp), encoded by a single UGG triplet, is the main reporter of intrinsic fluorescence properties of most natural proteins and peptides and is thus an attractive target for tailoring their spectral properties. Recent advances in research have provided substantial evidence that the natural protein translational machinery can be genetically reprogrammed to introduce a large number of non-coded (i.e. noncanonical) Trp analogues and surrogates into various proteins. Especially attractive targets for such an engineering approach are fluorescent proteins in which the chromophore is formed post-translationally from an amino acid sequence, like the green fluorescent protein from Aequorea victoria. With the currently available translationally active fluoro-, hydroxy-, amino-, halogen-, and chalcogen-containing Trp analogues and surrogates, the traditional methods for protein engineering and design can be supplemented or even fully replaced by these novel approaches. Future research will provide a further increase in the number of Trp-like amino acids that are available for redesign (by engineering of the genetic code) of native Trp residues and enable novel strategies to generate proteins with tailored spectral properties. PMID- 15551864 TI - Imprinted small RNA genes. AB - Genomic imprinting is an epigenetic phenomenon that results in differential expression of both alleles, depending on their parent of origin. We have recently identified many imprinted small non-coding RNA genes belonging to the C/D RNA and microRNA gene families, both of which are usually known to play key roles in post transcriptional metabolism of specific genes (e.g. C/D RNAs guide ribose methylation of target RNAs while microRNAs elicit either translational repression or RNA interference). Although the functional and evolutionary significance of this association between C/D RNA genes, microRNA genes and genomic imprinting is still highly elusive, these observations provide a framework for further analysis of the potential role of small non-coding RNAs in epigenetic control. PMID- 15551865 TI - Inter- and intra-molecular distances determined by EPR spectroscopy and site directed spin labeling reveal protein-protein and protein-oligonucleotide interaction. AB - Recent developments including pulse and multi-frequency techniques make the combination of site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy an attractive approach for the study of protein-protein or protein-oligonucleotide interaction. Analysis of the spin label side chain mobility, its solvent accessibility, the polarity of the spin label micro environment and distances between spin label side chains allow the modeling of protein domains or protein-protein interaction sites and their conformational changes with a spatial resolution at the level of the backbone fold. Structural changes can be detected with millisecond time resolution. Inter- and intra molecular distances are accessible in the range from approximately 0.5 to 8 nm by the combination of continuous wave and pulse EPR methods. Recent applications include the study of transmembrane substrate transport, membrane channel gating, gene regulation and signal transfer. PMID- 15551866 TI - Combined transport of water and ions through membrane channels. AB - The coupling of ion and water flow through membrane channels is under dispute. Among all human aquaporins only aquaporin-6 exhibits ion channel activity. Whether aquaporin-6 functions also as a water channel cannot yet be determined with confidence. Similarly, a comparison of single-channel water permeabilities from ion channels and aquaporins suggests that ion channels may play a secondary role as water channels. However, the fraction of absorbed fluid that crosses epithelial ion channels still remains to be determined. PMID- 15551867 TI - How do ABC transporters drive transport? AB - Members of the ATP-binding cassette (ABC) superfamily are integral membrane proteins that hydrolyze ATP to drive transport. In the last two decades these proteins have been extensively characterized on a genetic and biochemical level, and in recent years high-resolution crystal structures of several nucleotide binding domains and full-length transporters have extended our knowledge. Here we discuss the possible mechanisms of transport that have been derived from these crystal structures and the extensive available biochemical data. PMID- 15551868 TI - X-ray structure of fumarylacetoacetate hydrolase family member Homo sapiens FLJ36880. AB - The human protein FLJ36880 belongs to the fumarylacetoacetate hydrolase family. The X-ray structure of FLJ36880 has been determined to 2.2 A resolution employing the semi-automated high-throughput structural genomics approach of the Protein Structure Factory. FLJ36880 adopts a mixed beta-sandwich roll fold and forms homodimers in crystals as well as in solution. One Mg2+ ion is bound to each subunit of the dimeric protein by coordination to three carboxylate oxygens and three water molecules. These metal binding sites are accessible from the same surface of the dimer, partly due to the disorder of the undecapeptide stretch D29 to L39. The overall structure and metal binding site of FLJ36880 bear clear similarities to the C-terminal domain of the bifunctional enzyme HpcE from Escherichia coli C, fumarylacetoacetate hydrolase from Mus musculus and to YcgM (Apc5008) from E. coli 1262. These similarities provide a framework for suggesting biochemical functions and evolutionary relationships of FLJ36880. It appears highly probable that the metal binding sites are involved in an enzymatic activity related to the catabolism of aromatic amino acids. Two point mutations in the active-site of FAH, responsible for the metabolic disease hereditary tyrosinemia type I (HTI) in humans, affect residues that are structurally conserved in FLJ36880 and located in the putative catalytic site. PMID- 15551869 TI - Generation of glycogen- and albumin-producing hepatocyte-like cells from embryonic stem cells. AB - We present a novel two-step protocol for the differentiation of embryonic stem (ES) cells into the hepatic lineage. Differentiated hepatocyte-like cells express genes and proteins characteristic for endodermal and hepatic cells and acquire a functional hepatic phenotype as demonstrated by albumin secretion and glycogen storage. During differentiation, alpha-fetoprotein, albumin, transthyretin, alpha 1-antitrypsin, cytochrome P450 subunits 2b9 and 2b13 and tyrosine aminotransferase transcripts are upregulated. Quantitative RT-PCR data revealed a fetal hepatic phenotype corresponding to day 13-14 of liver development. Terminally differentiated hepatocyte-like cells show a bi-nucleated, cuboidal morphology labeled by albumin, alpha-1-antitrypsin, liver amylase, dipeptidyl peptidase IV, c-met and cytokeratin 18. ES-derived intermediate cell types transiently and partially co-express nestin with albumin and alpha-fetoprotein, respectively, but not cytokeratin 19. This finding suggests an ES-derived potential hepatic progenitor cell type, which is partially nestin-, albumin- and alpha-fetoprotein-positive, but cytokeratin 19-negative. PMID- 15551871 TI - A database search for double-strand containing RNAs in Dictyostelium discoideum. AB - In eukaryotic cells, double-stranded RNA is degraded to 21mers and triggers RNA interference. Using a pattern description language, we have searched the EMBL database for sequences with the potential to form double strands in cis in Dictyostelium discoideum. No extended inverted repeats were found in mRNAs. However, the antisense direction of some mRNAs encoding regulatory or developmentally regulated proteins showed the ability to form double-stranded regions. In EST archives, we found potential double strands derived from a few genes, but these transcripts are not continuously encoded in the genome. Most likely, they represent hybrid molecules of sense and antisense RNAs. PMID- 15551870 TI - Imaging morphological details and pathological differences of red blood cells using tapping-mode AFM. AB - The surface topography of red blood cells (RBCs) was investigated under near physiological conditions using atomic force microscopy (AFM). An immobilization protocol was established where RBCs are coupled via molecular bonds of the membrane glycoproteins to wheat germ agglutinin (WGA), which is covalently and flexibly tethered to the support. This results in a tight but non-invasive attachment of the cells. Using tapping-mode AFM, which is known as gentle imaging mode and therefore most appropriate for soft biological samples like erythrocytes, it was possible to resolve membrane skeleton structures without major distortions or deformations of the cell surface. Significant differences in the morphology of RBCs from healthy humans and patients with systemic lupus erythematosus (SLE) were observed on topographical images. The surface of RBCs from SLE patients showed characteristic circular-shaped holes with approx. 200 nm in diameter under physiological conditions, a possible morphological correlate to previously published changes in the SLE erythrocyte membrane. PMID- 15551872 TI - Segregation of partly melted molecules: isolation of CpG islands by polyacrylamide gel electrophoresis. AB - The technique of segregation of partly melted molecules (SPM) is a convenient and efficient method to isolate DNA fragments associated with CpG islands. The approach is conceptually simple and uses denaturant gradient gel electrophoresis to separate DNA molecules digested with restriction endonucleases. The SPM methodology has successfully been applied to the identification of genes from anonymous, unsequenced DNA fragments and CpG islands methylated in human cancer. In this article the theoretical background and practical application of the SPM method is reviewed. PMID- 15551873 TI - A monomeric mutant of restriction endonuclease EcoRI nicks DNA without sequence specificity. AB - We have mutated the monomer-monomer interface of the restriction endonuclease EcoRI in order to destabilize the homodimer and to stabilize heterodimers. Mutations of Leu158 to charged amino acid residues result in strong destabilization of the dimer. The largest effect was detected for the L158D mutant which is monomeric even at higher concentrations. It unspecifically degrades DNA by cleaving both single strands independently every 15 nucleotides on the average. Although cleavage is reproducible, it is not determined by nucleotide sequence but by general properties like conformation or deformability as has been found for other unspecific nucleases. Mutations of Ile230, which is in direct contact with Leu158 of the other subunit, cause structural changes with the loss of about ten percent alpha-helix content, but interfere only marginally with homodimerization and double strand cleavage. Again the mutation to aspartate shows the strongest effects. Mixtures of single mutants, one containing aspartate at one of the two positions and the other lysine at the corresponding position, form heterodimers. These are mainly stabilized compared to the homodimers by re establishment of the wild-type hydrophobic interaction at the not mutated residues while an interaction of aspartate and lysine seems energetically unfavorable in this structural context. PMID- 15551874 TI - [Rabies transmitted by vampires]. PMID- 15551875 TI - [Type 3 virus neuropathological findings in two human rabies cases in Bajo Baudo, Choco]. PMID- 15551876 TI - [Necrotizing fasciitis in an immunocompetent patient caused by Apophysomyces elegans]. AB - A case study is presented of a 7-year-old boy, seriously injured in a car accident, who developed a fatal infection due to Aphophysomyces elegans--a mold of the Mucoracea family. Fungal invasion was initially manifested by a spotted wound in the left lumbar region which developed into a necrotizing fasciitis. Later this progressed to the right lumbar area, including the gluteus and the corresponding flank. Antimycotic treatment proved ineffective, and the child died 8 weeks after the accident. Other cases due to this fungus are reviewed. PMID- 15551879 TI - [Effectiveness of treatments for hypertension in a sample of Colombian patients]. AB - Hypertension represents a high health cost because of its prevalence, its low level of diagnosis and control, and its role as a primary risk factor for other cardiovascular diseases. According to the JNC 7 report, hypertensive individuals have blood pressures of 140/90 mm Hg or higher; recommended treatment reduces these values to below 120/80 mm Hg. Co-morbidity and the presence of other risk factors must also be considered. In a random sample of 458 hypertensive patients from 6 Colombian cities, the effectiveness, tolerance and adherence to treatment was compared in cases with treatment of at least one year's duration. During routine blood pressure examinations, trained nurses obtained patient consent and additional anthropometric data, such as including co-morbidity, risk factors, antihypertensive medication prescribed, dosages and usage of unrelated medications. Some of the data were retrieved from the patients' medical histories. The average age of the patients was 57.6 +/- 13 years, with 67.5% women; 92% with complete adherence to the treatment and 59% not reporting adverse events associated with the medication. Forty-four percent were treated with antihypertensive monotherapy with the most commonly prescribed medications as follows (in order): hydrochlorothiazide, verapamil, enalapril, metoprolol and propanolol. Forty-five percent (n=207) were control patients, 35% were in a hypertensive stage 1 and 19.7% were in stage 2. Multivariate analysis showed that uncontrolled hypertension was significantly associated with geriatrics receiving a combination of antihypertensive medication and residence in three cities- Ibague, Barranquilla and Manizales--where smaller daily doses of hypertensive medications are prescribed. Health care teams are advised to adjust doses carefully to obtain clearly defined therapeutic objectives. PMID- 15551877 TI - [Molecular epidemiology of nosocomial infection by extended-spectrum beta lactamases-producing Klebsiella pneumoniae]. AB - Molecular epidemiology applied to the study of nosocomial infection has been fundamental in formulating and evaluating control methods. From patients in a level 3 Bogota hospital, Klebsiella pneumoniae samples were isolated that produced extended-spectrum beta-lactamases (ESBL). Each of 15 isolates was characterized microbiologically and by molecular characters realized by pulsed field gel electrophoresis (PFGE) and by repetitive-DNA sequences amplification (REP-PCR). Antimicrobial susceptibility and ESBL production was determined in accordance with NCCLS guidelines. The beta-lactamases were evaluated by isoelectric-focusing and PCR. Twelve (80%) of the isolates were associated with nosocomial infection; 11 of them were from intensive care units. The antibiotic susceptibility displayed 13 resistance patterns--87% presented co-resistance to amikacin, 53% to gentamicin, 33% to ciprofloxacin, 40% to cefepime, 67% to piperacillin/tazobactam, 60% to trimethoprim/sulfamethoxazole and 47% to chloranphenicol. All were sensitive to imipenem. Production of TEM and SHV beta lactamases was detected simultaneously in most isolates by isoelectric focusing and 93.3% produced a ceftazidimase of pl 8.2 of the SHV-5 type. The 15 isolates were grouped into 11 and 12 electrophoretic patterns by PFGE and REP-PCR, respectively. The degree of genetic variability indicated an endogenous origin of the nosocomial infections. PMID- 15551878 TI - [Expression and activity of polymorphisms in the 67-kDa protein of the NADPH oxidase system]. AB - The NADPH oxidase system plays a central role in the antimicrobial activity of phagocytes. This system is initiated by the translocation of cytosolic proteins p67phox, p47phox and p40phox to be in close contact with membrane flavocytochrome b558. This event begins the electron transfer from cytosolic NADPH to molecular oxygen to produce superoxide anions. Herein, a functional analysis is presented of p67phox polymorphisms identified from healthy humans. Mutations were generated in the p67phox cDNA by site-directed mutagenesis and then transiently expressed in COS7 cells that also expressed gp91phox, p22phox, and p47phox from stable transgenes. The changes Va1166lle, Pro329Ser and His389Gln correspond to possible polymorphisms identified in healthy individuals revealed a functional activity similar to COSphox cells transiently transfected with WT p67phox; therefore, these modifications are not associated with genetic deficiencies in NADPH oxidase. In conclusion, the COSphox system represents an easily transfectable model for analysis of NADPH oxidase function in intact cells. The analysis of mutant derivatives of p67phox provides insight into molecular mechanisms by which this subunit regulates the NADPH oxidase. PMID- 15551881 TI - [Exposure to Leptospira in stray dogs in the city of Cali]. AB - In Colombia, little information is available concerning the epidemiology of leptospirosis in urban environments. Furthermore, the role of dogs in the transmission cycle of leptospirosis in the urban setting is unclear. To explore the potential role of canines in the transmission of leptospirosis in Cali, a serological study was conducted with 197 serum samples collected from stray dogs during 2001 and 2003. Serum specimens were screened with the Microscopic Agglutination Test (MAT) and 7 serovars--Icterohaemorrhagiae, Canicola, Gryppotyphosa, Hardjo strain Hardjobovis, Pomona, Hardjo strain Hardjoprajitno and Bratislava. All serovars were provided by the Instituto Colombiano Agropecuario (ICA), Tulua, Colombia. The MAT was considered positive when 50% or more leptospiras were agglutinated with one or more serovars in a serum dilution of 1:100. At least one serovar showed evidence of infection in 41.1% of the dogs. The most prevalent serovar was Icterohaemorrhagiae, found in 55.6% of the seropositive dogs. 48.1% were co-agglutinations. No reactions against the serovars Pomona, Hardjo strain Hardjoprajitno and Bratislava were observed. These findings suggested that stray dogs are potential reservoirs of Leptospira in Cali and underscored the need to study the epidemiology of this disease in Colombia. PMID- 15551880 TI - [Molecular and biological characterization of the CIBMUQ/HDC strain, a reference strain for Colombian Toxoplasma gondii]. AB - There are few reports about characterization strains of Toxoplasma gondii that analyze the differences between isolates from Europe or United States with those obtained in South America. The current study analyzes virulence data from the mouse model, the gene SAG2 polymorphism by PCR-RFLP and microsatellite analysis in a single Colombian isolate. The strain was isolated from blood of a child with congenital toxoplasmosis, living in Armenia, Colombia. Analysis of virulence in the mouse showed that this strain has an LD100 of 10 tachyzoites. Both methods of genetic characterization demonstrated that this strain belonged to the clonal type 1 and was called HOM/CTCO/2002/CIBMUQ/BL/HDC (brief name: CIBMUQ/HDC). The CIBMUQ/HDC strain is the first Colombian strain available as a reference strain for national and international researchers. PMID- 15551882 TI - [Molecular surveillance of invasive penicillin-resistant Streptococcus pneumoniae Colombian isolates recovered from children less than 5 years of age]. AB - The rapid increase of penicillin-resistant Streptococcus pneumoniae isolates could be a consequence of the spread of clones or due to the antimicrobial selective pressure. The genetic relatedness of 190 invasive isolates S. pneumoniae with reduced susceptibility to penicillin recovered from Colombian children less than 5 years old during a surveillance study from 2000 to 2003 was determined by the use of pulsed-field electrophoresis (PFGE). Overall, 42 different PFGE patterns were identified, but 4 of them included 76% of all isolates. They were related with international clones 1-Spain23F, 2-Spain6B, 3 Spain9V and 26-Colombia23F. Our results indicated that the dissemination of penicillin-resistant S. pneumoniae was the result of the spread of international clones, specially, the 3-Spain9V clone. PMID- 15551883 TI - [Role of Langerhans cells in the immunity of leishmaniasis]. AB - Immune response induced against Leishmania parasites is influenced by several factors, one of the most important being the type of Antigen Presenting Cell (APC). Langerhans cells, a subpopulation of APC, are sentinel cells for detecting invader microorganisms; they reside in skin tissues at levels where the phlebotomine fly vector inoculates Leishmania parasites. Presence of microorganisms can induce activation of Langerhans cells, leading to their maturation and migration towards lymph nodes. There, Langerhans cells present antigens to T cells for their subsequent activation and specific differentiation into effector cells. Early after a Leishmania infection, few T cells have been observed at sites of infection, suggesting that infected macrophages have little opportunity to locate T cells specific for elimination of Leishmania parasites. However, Langerhans cells may be the cells available to provide signals for the stimulation of parasite-specific T-cell responses in the lymph node and for inducing T-cell migration to the infected skin. Herein, the main characteristics of Langerhans cells are reviewed, with special emphasis on their participation in cutaneous inflammatory response. The role of these cells in infections caused by protozoan parasites of the Leishmania genus is discussed. PMID- 15551884 TI - [Development of a record-keeping strategy for improvement of information retention in microbiological processing]. AB - The improvement of microbiological information processing in clinical laboratories depends on retention of information concerning who, what, when, how, and why each process was performed, the implementation of quality control procedures, and finally, its evaluation. The four objectives to be addressed are as follows: (1) to improve the collection of information concerned with microbiological processes, (2) to evaluate results of implemented strategies, (3) to offer a model data base to be used in research projects, and (4) to propose an evaluation model for comparative studies. To do this, microbiological cultures were collected from hospitalized patients from June 1997 to June 2003. Data for the analytical matrix were obtained from lab requests, medical history and the microbiological data. Statistical analyses were performed in Epi-Info 6. The laboratory records for 46,072 microbiological cultures were analyzed. Completion levels in data collection were compared between years 1997 and 2003. Samples from 1997 and 2003 showed 11% and 99% of the request forms specifically requesting microbiological culture, 11% and 99% were completed in 1997 and 2003, respectively. For the same years, 9% and 85% specifically stated the time of the request. Ten percent and 68%, respectively, provided complete information. Zero and 83% respectively stated who had collected the sample. Zero and 77%, respectively, specified the time of sample collection. Forms containing all relevent microbiological data were most complete with 78% and 96%, respectively. A database with 44 variables related to microbiological processes was created. In conclusion, improvement of microbiological data processing depends not only on the method of collection and completion of recorded information, but also on constant quality control and evaluation. PMID- 15551885 TI - [Cryptococcus spp. DNA extraction from environmental samples]. AB - The genus Cryptococcus encompasses 38 species, but only 3 are associated with disease in humans and animals, Cryptococcus laurentii, Cryptococcus albidus and Cryptococcus neoformans. The last one is the most frequently reported. The disease is acquired by the inhalation of infectious propagules present in the environment. The habitat has been established using extraction techniques with buffer supplemented with antibiotics and plating in selective media. The aim of this work was to evaluate several DNA extraction techniques for Cryptocococus spp. from environmental samples. The control isolates were C. neoformans, C. albidus, C. laurentii and Paracoccidiodes brasiliensis. We also used vermiculita and soil samples contaminated with different yeast concentrations (10 to 10(6) cells/g) and samples naturally contaminated with C. neoformans. DNA was extracted with physical and chemical methods and with a commercial kit, and the DNA was purified with agarose blocks and silica columns. For the PCR amplification we used the CN4-CN5 primers, which are specific for C. neoformans. Only the commercial kit allowed DNA extraction and amplification from contaminated soil samples up to a concentration of 10 cells/g and from one sample naturally colonized. With this work we extracted and amplified DNA from Cryptococcus spp. from environmental samples with appropriate PCR specificity, it will be a tool to establish the ecological areas of C. neoformans in our country. PMID- 15551886 TI - [Multibacillary leprosy recurrences]. PMID- 15551887 TI - [Pregnancy dengue]. PMID- 15551888 TI - When economy meets medicine--malignant hyperthermia in Taiwan. PMID- 15551889 TI - Nitrous oxide uptake in man: a new concept of uptake of inhalation anesthetics. AB - BACKGROUND: With proper recognition of the functional residual capacity (FRC) and the existence of the alveolar membrane in consideration of gas uptake, the uptake of nitrous oxide does not follow the previously described power function of time, 1,000 ml x t(-1/2). METHODS: By use of a simple subtraction method, nitrous oxide uptake was measured in patients undergoing anesthesia with nitrous oxide in oxygen and a high-dose narcotic for cardiac surgery. RESULTS: When FRC washin was considered separately, the uptake of nitrous oxide which started from zero, reached its peak value at the end of washin, and then diminished slowly. CONCLUSIONS: In contrast to previous publications, in which the time course of FA/FI was used to express the uptake of nitrous oxide, in spite of the area under the curve of FA/FI should not be used to express uptake, instead, the area above the curve, (1-FA/FI), fits the results and should be considered as representing uptake. PMID- 15551890 TI - The antinociceptive effect of buprenorphine and its long-acting ester buprenorphine benzoate in rats. AB - BACKGROUND: A long-acting analgesic is particularly desirable in patients with long-lasting pain. In order to prolong the duration of action of a potent analgesic, buprenorphine, a buprenorphine ester (buprenorphine benzoate) was synthesized in our laboratory. The aim of the study was to evaluate whether the ester form could have a long-lasting effect. METHODS: Two studies were carried out. In study 1, we evaluated the antinociceptive effect of buprenorphine HCI at dosage of 0.05, 0.1, and 0.6 micromol/kg. In study 2, we evaluated the antinociceptive effect of buprenorphine benzoate at dosage of 0.6 micromol/kg. Male Sprague-Dawley rats (n = 12 in each group of different treatment) were used and the antinociceptive effects of the testing compounds were evaluated using the plantar test (a thermal test). RESULTS: Intramuscular injection of buprenorphine HCI at dosage of 0.05, 0.1, and 0.6 micromol/kg in rats produced dose-related antinociceptive effects with duration of action of 3, 4, and 4 h, respectively. Intramuscular injection of buprenorphine benzoate at 0.6 micromol/kg produced a duration of action of 48 h. CONCLUSIONS: Intramuscular injection of buprenorphine benzoate in rats produced a longer duration of action, which is 12-fold longer than that of buprenorphine HCI. PMID- 15551891 TI - Frequent use of alternative airway techniques makes difficult intubations less and easier. AB - BACKGROUND: This study is to evaluate how alternative airway techniques work at different scenarios of difficult airways in our department where those techniques are popularized in daily practice. METHODS: This is an open and observational study. Ten thousand two hundred and twenty nine adult patients were included in the 7-month period of investigation. When a difficult intubation was recognized, the responsible anesthesiologist would complete a questionnaire that was designed to explore the possible causes relative to the difficulty, i.e. the laryngoscopic view grade, the muscle relaxant administered during induction, any difficulty of mask ventilation, the eventually successful intubation technique, and the recommendation on the next intubation to the same patient. RESULTS: About 22% of the patients received various airway techniques for intubation other than laryngoscopic technique in general anesthesia. Laryngoscopic intubation was planned firstly on 8,058 cases; the incidence of difficult intubation was 0.44% (36/8058). Laryngoscopic view was found to be grade 2 in 10 patients, grade 3 in 24, and grade 4 in 2 patients, respectively. In these difficult airway intubations were accomplished ultimately with a conventional laryngoscope in 13 patients, a lighting stylet in 20 patients, a laryngeal mask airway (LMA) in 2 patients, and surface tactile oral intubation (STOI) in 1 patient. As to these patients with difficult airways who would undergo general anesthesia with endotracheal intubation next time awake intubation was suggested only to 4 patients. CONCLUSIONS: In elective general anesthesia, frequent use of the alternative airway techniques, such as LMA for a surgical procedure of short duration and lighting stylet for a possibly difficult airway, will decrease the events of difficult intubations. Lighting stylet seems more advantageous than laryngoscope in management of difficult intubation caused by poor laryngoscopic view. PMID- 15551892 TI - A survey for prevention and treatment of malignant hyperthermia in Taiwan. AB - BACKGROUND: Malignant hyperthermia (MH) is a hypermetabolic disorder with high mortality. Intravenous administration of dantrolene is the specific treatment. However, the only means to lower mortality rate are early detection and suitable treatment. Therefore, properly monitoring and handy availability of dantrolene are essential to lower the mortality of MH. This study was designed to evaluate the capabilities in prevention and treatment of MH of the hospitals in Taiwan. METHODS: There were 102 certified training hospitals (including medical centers and regional hospitals) in Taiwan selected for this study. A questionnaire was designed to evaluate the capabilities of these hospitals in dealing with prevention and treatment of MH. RESULTS: A total of 66 copies of the questionnaire were returned, which gave a response rate of 64.7%. The results of the survey are as follows: Succinylcholine was frequently used for induction in 92% of the hospitals investigated. For monitoring during general anesthesia, pulse oximeter was routinely used in 66(100%) hospitals, and ETCO2 monitor in 51 (77.3%) hospitals but continuous body temperature was routinely monitored only in 13 (19.7%) hospitals. Six or more vials of dantrolene were stored and available for immediate use in 23 (34.9%) hospitals and the rest of 43 (65.1%) hospitals did not have any stock of dantrolene at all. Of these 43 hospitals, 25 (58.1%) relied on other hospitals to supply dantrolene and the time required to obtain dantrolene from other hospitals was 70.7 +/- 34.7 min. There were 21 cases who sustained MH in 17 (25.8%) hospitals in the past ten years, of whom 15 were resuscitated successfully and 6 died, thus giving a mortality rate of 28.6%. CONCLUSIONS: This study has revealed that in some hospitals in Taiwan intraoperatively (anesthetic) monitoring is inadequate and there is no stockpile of dantrolene for immediate treatment of MH. We recommend that appropriate anesthetic monitoring equipment should be strictly applied and a stock of initial dose of dantrolene (6 vials) should be kept, which are necessary for early diagnosis and treatment of MH. PMID- 15551893 TI - Supplementing epidural lidocaine with midazolam: effect on sensorymotor block level. AB - BACKGROUND: Midazolam, the only clinically available water-soluble benzodiazepine, has been reported to have an antinociceptive effect through neuroaxial pathway. Also it can be used epidurally for postoperative analgesia and chronic pain management. In a recent study it was disclosed that when midazolam was added to intrathecal bupivacaine it improved the duration and quality of the spinal anesthesia. In this study the effects of this drug on intraoperative motor and sensory blocks were evaluated. METHODS: We studied 53 adult ASA I and II patients scheduled for elective lower-limb surgery. All patients received epidural anesthesia with 19 ml of lidocaine and l ml additive. Patients were randomly allocated to three groups. Midazolam was administered epidurally in doses of 5 mg (1 ml) and 3 mg (1 ml) in groups land 2 respectively, and group 3 received normal saline Iml as placebo. Patients were evaluated for the onset time and duration of sensory and motor blocks, duration to the first rescue analgesic and hemodynamic changes. RESULTS: Onset of sensory block and time to peak effect was significantly faster and shorter in group one patients who received 5 mg midazolam as compared with those of other two groups (P = 0.002 and P = 0.001, respectively). Also, the durations of motor and sensory blocks were significantly longer in group one compared with those of other two groups (P = 0.004 and P = 0.045, respectively). CONCLUSIONS: Midazolam can improve the durations of sensory and motor blocks of lidocaine in a single epidural administration. It seems that the 5 mg dosage works better than the lower one. PMID- 15551894 TI - Transfusion-related acute lung injury--a report of two and the possible role of steroid. AB - Transfusion-related acute lung injury (TRALI) is a rare but potentially fatal clinical syndrome associated with transfusion. Dyspnea, hypoxemia, hypotension, and fever are the typical symptoms which mimic those of the adult respiratory distress syndrome (ARDS). The onset of TRALI is usually within 4-6 hours after transfusion. According to literature, TRALI is possibly underdiagnosed and underreported. Differential diagnosis includes fluid overload, allergy and sepsis. No definite protocol except supportive treatment is recommended up to now. We present two cases of TRALI and discuss the possible mechanisms and the role of steroid therapy. PMID- 15551896 TI - Pulmonary edema secondary to the airway obstruction in a patient with parkinson's disease. AB - Pulmonary edema is a potentially life-threatening complication of acute airway obstruction that develops rapidly without warning in persons who are otherwise well. Respiratory abnormalities have been reported in patients with Parkinson's disease, especially obstructive airway. To our knowledge, pulmonary edema secondary to airway obstruction in patients with Parkinson's disease has not been reported previously. Here, we describe a case of Parkinson's disease complicated with pulmonary edema, and airway obstruction might be the mechanism. PMID- 15551895 TI - Sudden onset of paraplegia after total hip replacement surgery in a patient with hepatocellular carcinoma--a case report. AB - Postoperative paraplegia is a major complication, of which the pathogenesis is usually multifactorial. This report is to discuss the case of a 36-year-old male patient who, after total hip replacement (THR), right, sustained a sudden-onset postoperative paraplegia. On subsequent examination, it was discovered that the patient had multiple vertebral metastases from hepatocellular carcinoma (HCC) resulting in thecal sac compression at L1 and S1 levels. This instance of distal spinal metastasis from HCC, with initial presentation of a sudden onset of paraplegia immediately after THR surgery, is worth reporting because of its being a rare occurrence and traumatizing effects on the patient, family members, and the surgical team. More importantly, we bring forth this case in order to advance an opinion concerning prevention of this devastating complication. Hence, we discuss the contributory factors and the appropriate perioperative survey and management relevant to cancer patients who are to undergo a non-cancer surgery. PMID- 15551897 TI - Occurrence of the Bezold-Jarisch reflex during Cesarean section under spinal anesthesia--a case report. AB - We report a 35-year-old parturient with gestational diabetes mellitus (GDM) history who sustained a nearly fatal Bezold-Jarisch reflex during Cesarean section under spinal anesthesia. A high spinal block combined with acute massive hemorrhage may produce life-threatening Bezold-Jarisch reflex which should be treated prophylactically and aggressively during surgery. Additionally, the autonomic disturbance associated with GDM might exaggerate the reflex in this episode. The patient survived after successful resuscitation and was discharged without any sequela. PMID- 15551898 TI - Fatal ischemia stroke in a patient with an asymptomatic carotid artery occlusion after lumbar spine surgery--a case report. AB - Recognizing carotid artery stenosis (CAS) in an asymptomatic patient preoperatively is difficult. We report a 63-year-old female who had diabetes mellitus and hypertension suffered from a fatal ischemic stroke secondary to a total occlusion of the left carotid artery after a lumbar spine surgery in the prone position. Intraoperative induced hypotension and neck rotation in this patient should be considered as contributory factors. According to previous studies extensively screening out CAS is not recommended. Using electroencephalography (EEG) or somatosensory evoked potential (SSEP) to monitor brain function and carefully positioning of the neck are recommended in these patients who are at risk of stroke while receiving induced hypotension during lumbar spine surgery in the prone position. PMID- 15551899 TI - The voice of male nursing. PMID- 15551900 TI - How to tackle verbal abuse. PMID- 15551901 TI - Breaking the bottlenecks. PMID- 15551903 TI - What you need to know about whooping cough. PMID- 15551902 TI - A competency framework for nurses in specialist roles. AB - As nurses undertake more specialist roles, strategies need to be in place to ensure competencies are maintained. University College London Hospitals has developed a competency framework to support personal and professional development of nurses in specialist roles. PMID- 15551904 TI - CVP monitoring. PMID- 15551905 TI - Treatment and management of acute appendicitis. AB - Acute appendicitis is one of the most common surgical conditions, and affects about seven per cent of the population. Perforation of the appendix and associated peritonitis is the most common complication. This guided reflection article discusses the signs, pre and postoperative treatment and nursing management of patients presenting with acute appendicitis. PMID- 15551906 TI - Improving the quality and appeal of pureed meals for patients. AB - This article describes how a critical incident led to a review of the pureed diet options offered to patients within a hospital setting. This resulted in significant changes within the trust, improving both the nutritional content and the appeal of the pureed diet menu. PMID- 15551907 TI - Using guidance to prevent errors when giving blood transfusions. PMID- 15551908 TI - Promoting health in the armed forces. PMID- 15551909 TI - The draft mental health bill as social policing. PMID- 15551910 TI - The heat is on. PMID- 15551911 TI - Heart attacks are predictable. PMID- 15551914 TI - Safe at the centre. PMID- 15551912 TI - Making more time. PMID- 15551913 TI - Sick and tired. PMID- 15551915 TI - Improving patient safety and incident reporting. AB - Patient safety is currently an international priority in health care, as it is widely accepted that the quality of healthcare provision, in terms of reducing errors and other forms of unnecessary patient harm, needs to be improved significantly. This article describes the work and position of the National Patient Safety Agency (NPSA) in NHS-funded care. It outlines the contribution made by two nurses who, as clinical specialty advisers (CSAs) in the organisation, are charged with helping to ensure that nursing issues are considered as an integral part of developing solutions to patient safety issues. PMID- 15551916 TI - Improving communication in day surgery settings. AB - This article emphasises the need for the development of a therapeutic relationship between nurse and patient in day surgery settings. While the short patient stay associated with day surgery may appear to mitigate against this, the use of interpersonal skills can directly support its augmentation. Enhancing skills such as listening, empowerment and caring has the potential to increase patient satisfaction with services in this area. PMID- 15551917 TI - Effective use of emollients in infants and young people. AB - Dry skin conditions are common in infants and young people. Emollients are the mainstay of treatment and play an important role in rehydrating dry skin. Nurses need to have a good knowledge of the causes of dry skin, the importance of skin barrier function and the properties and effective use of emollients if they are to help parents and children choose the most suitable product. PMID- 15551918 TI - The use of silver-based dressings in wound care. AB - Chronic wounds such as leg ulcers and pressure ulcers are often slow to heal. One of the causes of delayed wound healing is the presence of micro-organisms in the wound. A strategy for the prevention and treatment of wound colonisation or infection, which is receiving renewed attention, is the use of silver-based dressings. Silver has been used as an antimicrobial agent for centuries. It is effective against a broad range of bacteria (including methicillin and vancomycin resistant strains), yeast, fungi and viruses. A number of new silver-based dressings, some of which act by the sustained release of silver ions to the wound bed, have recently become available, but there are wide variations in the amount of data supporting the use of individual products. This article reviews the evidence base for silver-containing dressings to help practitioners select the most appropriate product for the type of wound being treated. PMID- 15551920 TI - The benefits of larval therapy in wound care. AB - This article discusses the role of larval therapy as an alternative to conventional treatment for acute or chronic wounds that respond poorly to conventional treatment (Courtenay 1999). Although not suitable for all wound types, this treatment is versatile and safe to use in hospitals and community settings (Davies 2004). In certain clinical situations, such as eliminating methicillin-resistant Staphylococcus aureus, larval therapy is considered to be a proven alternative in modern wound management (Brocklesby 2002). PMID- 15551919 TI - Exudate and care of the peri-wound skin. AB - A moist environment provides the optimum conditions for wound healing, but maintaining control of exudate and preventing its contact with the surrounding skin remain a challenge to carers. An essential part of management is identifying the underlying pathology of the wound. Correct diagnosis of the cause of increased exudate production is necessary for the implementation of appropriate interventions. Continual assessment of the wound and surrounding skin, together with evaluation of dressing performance, are integral to manage the different stages of wound healing. PMID- 15551921 TI - Take a second chance. PMID- 15551922 TI - Pharmacists and veterinary surgeons: cooperating on animal care. PMID- 15551923 TI - Veterinary nurses: undervalued and underpaid? PMID- 15551925 TI - Urinary corticoid:creatinine ratios in healthy pet dogs after oral low-dose dexamethasone suppression tests. AB - Eleven dogs were used in a trial to find a suitable dose of dexamethasone for an oral dexamethasone suppression test for the diagnosis of hyperadrenocorticism. Basal urinary corticoid:creatinine ratios were established in all 11 and then groups of seven were given oral doses of 0.02, 0.01 or 0.0075 mg dexamethasone/kg bodyweight and urine samples were collected at two-hour intervals from 08.00 to 22.00. The doses of 0.02 and 0.01 mg/kg consistently suppressed their urinary corticoid:creatinine ratios measured at 16.00 by a mean of more than 50 per cent and those of individual dogs to less than 1.0 x 10(-6), whereas the dose of 0.0075 mg/kg did not. PMID- 15551924 TI - Evaluation of baseline indices of sensibility in captive cetaceans. AB - Twelve possible tests of sensibility and six possible tests of vitality were evaluated for their ease of application and the reliability of the animals' responses in 25 animals of six species of captive cetaceans. The protocols for the application of the tests and the responses observed are described. PMID- 15551926 TI - Effects of a single dose of dexamethasone-21-isonicotinate on the metabolism of heifers in early lactation. AB - Eight Swedish crossbred heifers, about two-and-a-half years old, were given a single intramuscular dose of dexamethasone-21-isonicotinate between nine and 15 days after they had calved and eight similar heifers were left untreated. The treatment had no significant effects on the lipolytic activity of the heifers' fat tissues, and no effect on the concentrations of non-esterified fatty acids and beta-hydroxybutyrate in blood or the triacylglycerol content of the liver. However, there were significant increases in plasma glucose concentrations two days after the injection and in plasma insulin concentrations two and four days after the injection. PMID- 15551927 TI - Distribution of PrP(res) in the brains of BSE-affected cows detected by active surveillance in Catalonia, Spain. PMID- 15551929 TI - Successful treatment of megabacteriosis in a canary (Serinus canaria) with nystatin. PMID- 15551928 TI - Enzootic nasal tumour virus demonstrated in sheep in Senegal by direct detection of provirus from tumour DNA. PMID- 15551930 TI - Ultrasonographic diagnosis of omasal leiomyoma in a cow. PMID- 15551931 TI - Effect of sow parity on the transmission of Lawsonia intracellularis to piglets. PMID- 15551932 TI - Spinal cord lesions in broiler chickens. PMID- 15551933 TI - Changes to TB testing. PMID- 15551934 TI - Early neutering of cats. PMID- 15551935 TI - Early neutering of cats. PMID- 15551936 TI - A cumulative review of the range and incidence of significant adverse events associated with acupuncture. AB - OBJECTIVE: To summarise the range and frequency of significant adverse events associated with acupuncture in order to provide evidence on which to base continuing efforts to improve the safety of acupuncture practice. METHODS: Searches were conducted of computerised databases, previous reviews of case reports, population surveys, prospective surveys of acupuncture practice and relevant sections of textbooks for primary and secondary reports to indicate the range of significant adverse events associated with acupuncture. Data from prospective surveys of acupuncture were combined to estimate the incidence of serious adverse events. RESULTS: A total of 715 adverse events was included. There were 90 primary reports of trauma, and 186 secondary reports; the most common were pneumothorax and injury to the central nervous system. Infection accounted for 204 primary reports and 91 secondary reports. Over 60% of these cases were hepatitis B. The next most common infection was of the external ear, as a complication of auricular acupuncture. The 144 miscellaneous events mainly comprised seizures and drowsiness judged severe enough to cause a traffic hazard. There were 12 primary reports of deaths. According to the evidence from 12 prospective studies which surveyed more than a million treatments, the risk of a serious adverse event with acupuncture is estimated to be 0.05 per 10 000 treatments, and 0.55 per 10 000 individual patients. CONCLUSIONS: The risk of serious events occurring in association with acupuncture is very low, below that of many common medical treatments. The range of adverse events reported is wide and some events, specifically trauma and some episodes of infection, are likely to be avoidable. PMID- 15551937 TI - BMAS policy statements in some controversial areas of acupuncture practice. AB - The safe and competent practice of acupuncture requires the practitioner to conform to a number of basic principles. These include minimising the risk of transmission of infection, maintaining sufficient knowledge of anatomy to guide safe needling, and seeking an orthodox medical diagnosis before embarking on treatment. Beyond these basic principles, there are certain circumstances in practice that generate regular debate amongst practitioners. This article details advice on a limited selection of such circumstances. This advice has been drawn from a set of policy statements originally drafted to facilitate clarity and consistency within the teaching of Western medical acupuncture provided by the BMAS. It is not comprehensive, but covers the areas where there were the greatest differences in approach among the teaching staff. The original policy statements were also incorporated into the BMAS Code of Practice. By using the guidance in this article, in addition to applying sound clinical judgement and a knowledge of relevant anatomy, we hope that practitioners will minimise both the theoretical risks and the reported serious adverse events related to acupuncture, yet be able to practise unencumbered by illogical restrictions. PMID- 15551938 TI - A survey of the use of self-acupuncture in pain clinics--a safe way to meet increasing demand? AB - An acupuncture service is well established within a pain clinic in Nottingham, England, and is now unable to meet the increasing demand for treatment despite recent expansion. Patients used to be offered training in self-acupuncture. This was withdrawn because of safety concerns, but is being considered again as a way of meeting the demand. There is little published research on the topic, so a survey of 42 English hospitals was conducted to establish whether acupuncture services are provided, and to discover whether others were offering training in self-acupuncture to patients. Thirty hospitals replied, 23 of which offer an acupuncture service. The average waiting time for the first acupuncture treatment was 18.5 weeks, and the average waiting time for follow up treatments was nine weeks. One hospital taught patients auricular self-acupuncture, another was planning to teach patients, and a third hospital had previously taught patients but stopped. Discussing these findings, concerns are raised about the safety of self-acupuncture, and issues about patient selection, training, information, supervision and supply of materials are reviewed. A debate on these issues would be valuable. PMID- 15551940 TI - Debate--patients should be encouraged to treat themselves. AB - In favour of self-acupuncture, this is a useful way to prolong the effects of acupuncture when the response is only brief or patients cannot attend frequently. If the patient is capable and the condition is suitable for self-acupuncture, patients can be taught how to do it and then seen for review. Patients should be provided with complete information sheets, and one example is presented. Against self-acupuncture, patient safety is paramount and would be compromised by teaching them how to do acupuncture. In addition, serious accidents have happened with self-acupuncture, including a death. Other arguments against it are that it is less effective than standard acupuncture, the patient misses out on the therapeutic relationship, and safer alternative methods exist. PMID- 15551941 TI - Acupuncture for chronic neck pain--a cohort study in an NHS pain clinic. AB - The study investigates the outcome of acupuncture for chronic neck pain in a cohort of patients referred to an NHS chronic pain clinic. One hundred and seventy two patients were selected for acupuncture over a period of 6.5 years. Treatment was given by a single acupuncturist and consisted of a course of needle acupuncture for an average of seven sessions per patient. Treatment outcome was measured by an oral rating scale of improvement at the end of treatment and at follow up six months and one year after treatment. Nineteen patients were withdrawn from treatment for various reasons, two for adverse events. One hundred and fifty three patients were evaluated, of whom 68% had a successful outcome from acupuncture, reporting an improvement in pain of at least 50%. The success rate was higher in patients with a short duration of pain: 85% in patients with pain for up to three months and 78% with pain for up to six months. Long-term follow up showed that 49% of the patients who completed treatment had maintained the benefit after six months, and 40% at one year. The results indicate that acupuncture can be an effective treatment for selected patients with chronic neck pain. PMID- 15551942 TI - Unilateral septic arthritis of a lumbar facet joint secondary to acupuncture treatment--a case report. AB - This report describes a case of septic arthritis of the lumbar facet joint probably as a result of acupuncture treatment. A 48 year old man with a long history of back pain presented with a two week history of increasing pain following a third session of acupuncture. Examination revealed tenderness in the right lumbosacral area and laboratory investigations revealed raised inflammatory markers with negative blood cultures. A bone scan and MRI scan showed evidence of septic arthritis of the right L5/S1 facet joint. An x ray computed tomography guided biopsy was carried out which isolated staphylococcus aureus. The patient was initially treated with intravenous antibiotics. A repeat MRI scan demonstrated persistent septic arthritis with adjacent early abscess formation. Surgical debridement of the facet joint was therefore performed. The patient had resolution of his symptoms and the inflammatory markers returned to normal. He regained a full range of movement of the lumbar spine. Very few cases have been reported of lumbar facet joint septic arthritis and this condition is rare in association with acupuncture treatment. A high index of suspicion needs to be maintained and if conservative management fails then debridement can result in an acceptable outcome. PMID- 15551943 TI - Polar acupuncture. AB - Musculoskeletal disorders are common in people who undertake adventure travel to the Antarctic, and in those who support them, because of the hard physical demands and lack of rest. This paper describes the successful use of acupuncture as first line treatment for ten patients in these circumstances, and comments on its advantages, particularly in its capacity to reduce the use of non-steroidal anti-inflammatory drugs. PMID- 15551944 TI - Report on acupuncture in patients with valvular heart disease and prosthetic valves--the opinion of COPE. PMID- 15551945 TI - [Occupational exposure to endocrine disruptors: state of the art]. AB - Research into how exposure to "endocrine disrupters chemicals" affects human health is attracting increasing attention among European and international scientists since these contaminants are so widespread in the home and work environment and can have far-reaching effects on mental and physical health. Here we draw a general picture of studies to date on specific occupational exposures to single chemicals such as bisphenol A, styrene, etc., or homogeneous groups such as pesticides, metals, dioxins, phthalates and others. Although the exposure occurs in different ways, the toxic mechanisms of action vary widely, and it is hard to establish precisely the conditions of occupational exposure, significant correlations are nevertheless evident between the potential dose and its effects and further studies are certainly needed. There is still much debate on the epidemiological methods employed, which may overestimate exposure. The "measure" or at least an accurate description of exposure conditions is critical to the whole question and attempts to ensure this involve standardized procedures and statistical tests as the basis for a protocol for assessing the risk of occupational exposure. Investigations to date have focused on the effects on the reproductive system, in males in particular. However, considering the broad range of equilibria and systems on which endocrine destructive compounds can act, the international scientific community needs to persist in its efforts to develop methods for checking the effects on other endocrine organs--particularly the thyroid gland--and on the immune and neurological systems. PMID- 15551946 TI - [Vibration, back pain and physical exercise in high-risk professionals: a cross sectional study]. AB - Repeated loads and vibration stress in professional settings are relevant risk factors for back pain. Aim of this cross-sectional study was to estimate: a) the prevalence of back pain in two high-risk professional samples (helicopter pilots and bus drivers); b) the association between physical/sports exercise and back pain subjective perception across age. Prevalence of back pain is 94% in helicopter pilots and 74% in bus drivers; prevalence of back pain significantly increases with age. The positive effect of regular physical/sports exercise on subjective back pain significantly decreases with age. Physical or sports exercise adapted to structural characteristic of patients can result effective in diminishing personal impairment in subjects at professional risk. PMID- 15551948 TI - [Judgment on work capacity of a health worker with schizophrenia]. AB - We show the case of a worker affected with schizophrenia who was declared by the psychiatrist fit to work. The "Medico Competente" on the contrary, on the basis of his studies and in particular on his deep knowledge of the features of the office and the place where that worker works, decides that he is not fit to do that job. This judgement is confirmed by the "ASL-Collegio Medico". The case we described emphasizes in a very strong way what is written in the item 17, paragraph 1, letter a of the D.Lgs 626/94, which extends the judgement of the fitness to work given by the "Medico Competente" even to the psychic sphere of the worker. PMID- 15551947 TI - "In vitro" effects of different arsenic compounds on PBMC (preliminary study). AB - Aim of this investigation was to compare the effects of 10(-4) M and 10(-7) M As compounds on spontaneous and PHA stimulated PBMC proliferation and IFN-gamma and TNF-alpha release. The inhibitory effect of the 10(-4) M As salts was in the following order: momo-methyl-arsinous acic (MMAs(III)) > sodium arsenite (As(III)) > tetraphenyl arsonium chloride (As(V)) > sodium arsenate (As(V)) > potassium- and sodium-esa-fluorum arsenate (As(V)) > dimethyl arsinic acid (DMAs(V)), while monomethyl-arsonic-acid (MMAs(V)) and arsenobetaine did not exert immune effects. 10(-7) M MMAs(III) stimulated the spontaneous PBMC proliferation, while As(III) and DMAs(V) enhanced the PHA stimulated PBMC proliferation. This study shows that the immune effects of As salts depends on speciation; moreover, the immunotoxicity of inorganic arsenic in part depends on the intracellular bio-synthesis of MMAs(III) from MMAs(V). PMID- 15551949 TI - [Noise-related occupational risk aboard fishing vessels: considerations on prevention and the protection of exposed workers]. AB - Recent legislation regarding the safety of workers aboard fishing vessels requires the appointment by ship owners of a Reference Physician in charge of health surveillance, preventive inspections and related tasks. As maritime workers, especially fishermen, have always been excluded from legal protection of occupational health, there are no exhaustive data on the incidence of their occupational disease. Several epidemiological studies of fishermen have evidenced a high prevalence and incidence of occupational conditions, among which noise related hypoacousia. We report data of a phonometric survey conducted aboard six fishing vessels carrying a crew of less than six fishing in the mid-Adriatic. Measurements were performed during fishing and navigation aboard five vessels fitted with a fixed-pitch propeller and during fishing only aboard one vessel fitted with an controllable pitch propeller. Measurements were conducted: 1) in the engine rooms; 2) in the work area on deck; 3) at the winch; 4) in the wheelhouse; 5) in the mess-room and kitchen; 6) in the sleeping quarters. Results show that the equivalent sound pressure level in the engine rooms consistently exceeded 90 dBA on all vessels. The speed of the vessels fitted with the fixed pitch propeller is 3-4 knots in the fishing phase and around 10 knots during navigation to and from the fishing grounds; noise emission is lower with the former regimen because of the smaller number of engine revolutions per minute. Our survey demonstrated considerably different noise levels in the various areas of vessels. One key element in workers' exposure, the tasks assigned and the environmental working conditions is of course the type of fishing in which the vessel is engaged. Further phonometric studies are required to assess the daily level of exposure per crew member, which represents the reference for the noise related risk of each subject. Knowledge of the sound pressure levels in the work environment and the length of daily exposure of each crew member will allow to assess the level of occupational exposure and consequently enact the proper prevention and protection measures by the Reference Physician. PMID- 15551951 TI - [Echinococcosis: a persistent endemic]. AB - Echinococcosis seems to be still endemic in Italy. Data from epidemiological studies in middle- south Italy indicates that this parasitosis is often a work- related disease. Our survey concerning findings among patients from surgical departments in Tuscany confirms the rsults of the studies. Echinococcosis is still endemic and dairy farming seems to be a risk factor. PMID- 15551950 TI - [Oxidative stress in station service workers]. AB - INTRODUCTION: The aim of this study is to identify an oxidative stress in service station workers. Previous studies verified an increased incidence of leukemia and myeloma, however other authors haven't verified it. There are reports of nasal, pharyngeal, laryngeal, and lung cancer in service station workers. Our study wants to evaluate the oxidative balance in the fuel workers. MATERIAL AND METHODS: We studied 44 subjects with gasoline exposure and 29 control subjects. We determined the blood concentrations of Glutathione reduced and oxidized, Protein sulfhydrylic (PSH) Vitamine E, Vitamine C, Malondialdehyde, Protein oxidized (OX-PROT) and beta carotene. The t test was performed to analyze the differences between the means, the Chi square was used to evaluate the statistical significance of associations between variable categorical (redox index). The Anova test excluded the confusing effect of age, smoke and alcohol habit. RESULTS: The mean age of the workers was 36.6 years, instead the control group was 38. In the workers Glutathione reduced, Vit. E and Beta carotene were lower than in the control subjects, this difference was statistically significant (p < 0.01). The Malondialdehyde concentration was higher in the workers higher than in the control group, but this difference wasn't statistically significant. DISCUSSION AND CONCLUSIONS: Our data demonstrated Glutathione, Vit. E, and Beta carotene are useful to verify a reduction of the antioxidant activity. The only marker of the presence of oxidative injury that correlated to work exposure was the malondialdehyde. The redox index was surest marker. The limit of our study is the number of control group, it was little and lower than workers. Conclusively we believe it's useful to continue our studies and, if our results are going to be confirmed, we retain that stress oxidative determination would be verified in occupational medicine using these markers, especially to study exposure of the fuel workers who were investigated less and, in our opinion, would receive more attention. PMID- 15551952 TI - [Excretion kinetics of phenylhydroxyethyl mercapturic acids (PHEMAs), ethanol consumption, and chronic exposure to styrene: preliminary data on humans]. AB - Styrene (S) is a widely used aromatic hydrocarbon, responsible for several adverse effects. In humans, the metabolism of S is well characterized: besides the major metabolites (mandelic and phenylglyoxylic acid), a minor metabolic pathway leads to phenylhydroxyethyl mercapturic acids (PHEMAs) [N-acetyl-S-(1 phenyl-2-hydroxyethyl)-L-cysteine (M1) and N-acetyl-S-(2-phenyl-2-hydroxyethyl)-L cysteine (M2)], that are potentially useful for biomonitoring purposes. A pilot study on a volunteer exposed under controlled conditions to S, with or without ethanol administration, allowed us to characterize the excretion profile of PHEMAs and the ethanol-induced interference on PHEMAs metabolic pathway. We further considered a group of 9 workers exposed to S during the working week to determine the confounding role of chronic exposure. Our results confirm the wide interindividual variability of both the biotransformation rate of S into PHEMAs and of the excretion rate of these metabolites. Moreover, both the above parameters changed during the working week, suggesting the existence of a large intraindividual variability as a consequence of the exposure to S and to other solvents. As a practical rule, the data indicate that it is necessary to collect samples at the beginning of the working week when studies on the correlation between genotype and phenotype are carried out. Finally, the results emphasise the importance of excluding an even extemporary ethanol assumption when practicing a biological monitoring programme based on the determination of urinary PHEMAs. PMID- 15551953 TI - [Comparative analysis of the use of 4 methods in the evaluation of the biomechanical risk to the upper limb]. AB - In the last years notices of Work Related Musculo-Skeletal Disorders are significantly increased, for a much widespread mechanism of disease and risk identification. About the last, the methods of assessment, even if in great number, are not universally recognized. This study reports the application of four methods (Check list OCRA, OREGE, Strain Index, ACGIH) to evaluate 12 work emplacements at a different biomechanical overload level. The comparison revealed variability between the methods in situations of middle risk, while the methods substantially gave similar results in situations without any risk or with high risk. Methods could not reveal every component of the risk in a reproducible and univocal way also when the risk assessment was accordant. It is necessary a following analytical evaluation of each risk factor contributing to define the sintetic index. In the complex of biomechanical risk factors these ones might not be considered able to define unequivocally "accettable" or "not accettable" components. PMID- 15551954 TI - [De Quervain's tenosynovitis and work with hand tools: a case study]. AB - Work-related musculoskeletal disorders of the upper limbs are a frequent complaint and their pathogenetic mechanism is often related to mechanical overload of the body region involved without traumatic events. The occupational origin of such disorders is much debated, since the clinical pictures are often not specific and may be found in subjects who do not perform hazardous work tasks. A better understanding of the relationship between clinical symptoms, diagnosis, and functional capacity would provide a deeper knowledge in decision making for return to work and rehabilitation strategies. In this paper we present a case study of a worker using scissors who developed De Quervain's syndrome in the dominant hand. A methodology for the biomechanical investigation of the work task and the possible occupational aetiology of the clinical picture are discussed according to the findings of the specific literature. PMID- 15551955 TI - [Drug addiction and working]. AB - The majority of subjects with drugs addiction is actively working. Therefore, the possible consequences of drug addiction in the workplace are presented in this paper. They entail reduction of work performance, increase of accidents and absenteism. The role of the occupational physician is also discussed in terms of drug addiction diagnosis and job fitness evaluation. The opportunity to use drug testing in the workplace is also considered, for those cases of occasional drugs abuse. PMID- 15551956 TI - [Neurobehavioral tests in occupational medicine: diagnosis and prevention]. AB - Modern neurobehavioral methods are used in the assessment of the early effects due to exposure to neurotoxic agents in working and general environment. This paper describes the history and evolution of neurobehavioral methods and their contribution for research on the effects due to the exposure to neurotoxic substances. Moreover, the paper describes the different fields of application of neurobehavioral tests: experimental, epidemiological and follow-up studies, health surveillance, clinical diagnosis, definition of job fitness and risk assessment for the definition of threshold limit values. PMID- 15551957 TI - [Alcohol and working]. AB - Due to its negative impact on both health and productivity, alcohol misuse is a serious concern in the workplace. Some occupations (e.g. employees of the catering and hotel trade, seamen, sales representatives, brewers and distillers, journalists, physicians, lawyers) are associated with a high rate of alcohol abuse. Alcohol intake can modify worker's behaviour (impaired judgement and vigilance, dulled reflexes) causing reduced performance, mistakes during operating procedures, accidents and injuries. Moreover it can affect the toxicokinetic and toxicodinamic properties of several substances in the workplace, inducing a more complex evaluation of exposure assessment and diagnostic procedures of occupational diseases. The occupational physician, during health surveillance program, can face several alcohol related issues. These entail diagnostic evaluation of alcoholism, job fitness evaluation, in heavy drinkers, advise of rehabilitation and health promotion program. PMID- 15551958 TI - [Analysis on intraspecific variation of Cnidium monnieri by RAPD]. AB - OBJECTIVE: To study the genetic variation of Cnidium monnieri from different regions. METHODS: Random amplified polymophic DNA technique was used to analyze genetic polymorphy of Cnidium monnieri from 9 different regions, and dendrogram was constructed by UPGMA. RESULTS: 20 random primers were used for polymorphic selection. A total of 75 bands were amplified, 64 of which were polymorphism. CONCLUSION: The smaller the geographic distances between two Cnidium monnieri, the smaller genetic differences. However, it should not be overlooked that natural environment, especially the local ecological conditions may also show some influences on the genetic variations among different C. monnieri samples. PMID- 15551959 TI - [The dynamic change of total alkaloids content from Pinellia ternate]. AB - OBJECTIVE: To determine the alkaloids content of Pinellia ternate in different growing time for finding out the best harvest time. METHODS: The alkaloid was extracted by chloroform from Pinellia ternate and tested at 417 nm with the spectroscopy by acid dye. The result was analyzed in One-Way ANOVA. RESULTS: The calibration curve of alkaloid was linear( r = 0.9989). The recovery rate was perfect (100.1%). The alkaloids content in different growing time had very significant difference ( F = 12.789, P < 0.01). CONCLUSION: This method is simple and quick. The alkaloids content in whole seedling stage is higher than that in other growing stages, while the mean total alkaloids output per plant is the highest in intensive seedling-downfallen stage which is considered as the best harvest time. PMID- 15551960 TI - [Studies on distribution of paris and annual growing characters of some species on Mountain E' mei]. AB - The research of distribution and habitat of Paris on Mountain E' mei showed that: (1) there were 8 species and 3 varieties of Paris on Mt. E' mei, most of them distributed from alti. 1000m to 1400m in humid and fertile, neutral or acid soil under the evergreen broad-leaved forests, bamboo forests and shrub forests. (2) only Paris bashanensis distributed at higher altitude about 1800 approximately 2800 m; (3) the growing time of P. polyphylla var. chinensis and other two species planted at the lower E' mei mountain area, was later and shorter than those plantd at Kunming; (4) the whole rhizome or with terminal bud as the planting material had a high survival rate and grew fast. PMID- 15551961 TI - [Application of allele-specific primer in the identificatin of Hedyotis diffusa]. AB - OBJECTIVE: To develop a convenient and effective method for the identification of Hedyotis diffusa. METHODS: DNA templates were extracted from H. diffusa and its adulterants samples on commercial markets. And DNA fragments of rDNA ITS-2 regions were amplified and sequenced subsequently. ITS-2 sequences of all samples were aligned. The allele-specific primer was designed for distinguishing H. diffusa. RESULTS: The nucleotide difference between H. diffusa and 8 other species is obvious in the ITS-2 region. In addition, the allele-specific primers were employed to amplify the DNA from H. diffusa and 8 other species. The result indicated that a 392 bp DNA fragment was amplified from H. diffusa, whereas no any fragment was amplified from 8 other species under the same reaction condition. CONCLUSION: The primers designed in the present study were highly specific for H. diffusa. They could be used as key components in the H. diffusa identification kit. PMID- 15551962 TI - [Isolation and identification of minor bioactive saponins from the leaves of Panax notoginseng]. AB - The minor bioactive saponins of the total saponin from the leaves of Panax notoginseng (Burk.) F. H. Chen. were separated and purified by column chromatographies on silica gel and thin layer gel. Four compounds were identified as: ginsenoside C-K (I) [20 (S) -protopanaxadiol 20-O-beta-D-glucopyranoside], ginsenoside-Rh1 (II) [20 (S) -protopanaxatriol-6-O-beta-D-glucopyranoside], ginsenoside-Mc (III) [20 (S)-protopanaxadiol 20-O-alpha-L-arabinofuranosyl (1 --> 6) -D-glucopyranoside] and notoginsenoside-Fe (IV) [20(S) -protopanaxadiol-3-O beta-D-glucopyranosido-20-O-alpha-L-arabinofuranosyl(1 --> 6)-beta-D glucopyranoside]. Compound I was isolated from the leaves of Panax notoginseng for the first time. PMID- 15551963 TI - [Studies on the constituents from Sambucus chinese]. AB - Two compounds were isolated from the petroleum ether and ethyl acetate soluble fraction of ethanolic extract from Sambucus chinese Lindl. They were identified as: 3-beta-acetoxyolean-12-ene (I) , ethyl caffeate (II). I and II were isolated from this plant for the first time. PMID- 15551964 TI - [Study on flavonoids from stem bark of Pongamia pinnata]. AB - Seven flavonoids, pongaflavone (1), karanjin (2), pongapin (3), pongachromene (4), 3,7-Dimethoxy-3', 4'-methylenedioxyflavone (5), millettocalyxin C( 6), 3,3',4', 7-tetramethoxyflavone (7), were isolated from 50% EtOH syrup of the bark of Pongamia pinnata and structureal elucidated on the base of spectral data. Compound 6 was isolated for first time from the plant of the genus Pongamia. PMID- 15551965 TI - [Determination and comparision of gentiopicroside of various parts of five species of genus Gentiana collected from Xinjiang by TLCS]. AB - OBJECTIVE: To study gentiopicroside's determination and comparison of various parts of herb by TLCS and make it impossible to uncover resource of Gentian in Xinjiang area. METHOD: Gentiopicroside's determination and comparison of various parts of herb in Gentiana Plants were carried out by TLCS. RESULT: The average recovery rate was 98.66% (RSD = 1.33%, n = 5). The Contents were stable. The results kept a high degree of comparable with HPLC ones. CONCLUSION: This method was good-steady, high-accurate, simple, convenient, sensitive, reproducible. PMID- 15551966 TI - [Effects of osthol on androgen level and nitric oxide synthase activity in castrate rats]. AB - OBJECTIVE: To investigate the effects of osthol on androgen level and nitric oxide synthase (NOS) activity in castrate rats. METHODS: Immature castrate male rats were treated with osthol daily for 20 d. The levels of testosterone(T), luteinizing hormone (LH) and follicle stimulating hormone (FSH) in serum and the activity of NOS, the nitric monoxide (NO) level in penis tissue were determined. RESULTS: T( P < 0.05), LH( P < 0.05) and FSH( P < 0.01) were significantly increased at the high dose of osthol treatment. The high and low dose of osthol treatment significantly enhanced the activity of NOS (P < 0.01). CONCLUSION: Osthol could increase androgen, gonadotropin and nitric oxide synthase (NOS) activity. It suggested that osthol had androgen-like effect and gonadotropin-like effect. PMID- 15551967 TI - [Effects of Capsule Yizhi on the delayed neuronal death in hippocampal CA1 region and memory function after global cerebral ischemia in rats]. AB - OBJECTIVE: To observe the effects of Capsule Yizhi on the delayed neuronal death in hippocampal CA1 region and memory function after global cerebral ischemia in rats. METHODS: The model of acute reperfusion injury after cerebral ischemia in rats was made by vertebral and carotid arteries occlusion. 1,2,4,8 and 40 days later, the neurons in hippocampal CA1 region were immunohistochemically stained and calculated. And the Morris water maze test was carried out to observe the learning and memory capacities in the 40th day. RESULTS: The number of normal neurons was significantly higher in Capsule Yizhi (100 mg/kg) group than that in the control group. Compared with the control group, Capsule Yizhi (100 mg/kg) could significantly shorten time in finding the platform under the water surface in Morris water maze test. CONCLUSION: Capsule Yizhi had protective effects on neurons in hippocampal CA1 region,and presented promising effects on learning and memory dysfunction after global ischemia/reperfusion injury. PMID- 15551968 TI - [The effects of extracts from Herba Lagopsis on lymph microcirculation of acute blood stasis rats]. AB - OBJECTIVE: To observe the effects of the extracts from Herba Lagopsis (HLE) on lymph microcirculation of acute blood stasis rats. METHODS: The acute blood stasis model was made by iv dextran (10 ml/kg) within 1 min. After 6 minutes, HLE(1.0g/kg) was given intravenously to the treatment group and the same volume of NS was done in the normal control group. The changes of contractility of mesenteric micro-lymphatics (ML) was observed by using a vital microscope with TV recorder. RESULTS: HLE could markedly improve the ML spontaneous constraction frequency, lymphatics constractive activity (Index I), total lymphatics constractive activity (Index II) and lymphatics dynamics (L. D-Index) (P < 0.05 approximately 0.01). CONCLUSION: HLE played an important role in lymph microcirculation of acute blood stasis rats. PMID- 15551969 TI - [Experimental study on preventing postoperative posterior urethra scar in response to Tablet Fu Kang]. AB - OBJECTIVE: To find an effective measurement to prevent and decrease the posterior urethral scar stricture. METHODS: Twenty New Zealand rabbits were chosen. All the rabbits were made posterior urethra disruption model, and at the same time, the urethra anastomosis operation was performed. Twenty rabbits were divided into two groups after operation at random. One group was treated by Tablet Fu Kang (experimental group), another group was treated by NS (control group). All the rabbits were killed three weeks later, the urethra anastomotic stoma tissue was chosen and stained with Masson. All the figures of slides were inputted into figure analytic system. RESULTS: There were little collagenous fiber, sparsely and arranged in good order in scar tissue of urethra anastomotic stoma in experimental group. The area of collagenous fibers was 117.11 +/- 10.17 microm2 in three visual fields. There were many collagenous fibers, densely,and arranged in disorder in scar tissue of urethra anastomotic stoma in control group, the relative area was 136.43 +/- 15.85 microm2. There was statistic significance when comparing the area of collagenous fiber in two groups (P < 0.05). CONCLUSIONS: Tablet Fu Kang could restrain the hyperplasia of fibroblast in the process of wound healing of rabbit smooth muscle in urethra. PMID- 15551970 TI - [Study on optimizing the extraction process of total free organic acids in Pinellia ternate Breit]. AB - OBJECTIVE: To optimize the extraction process of total free organic acids in Pinellia ternate Breit. METHOD: The optimum extraction was investigated by total free organic acids determined by direct potential titration. The concentration of ethanol, amount of ethanol, extraction time and extraction times were the four factors in the experiment. RESULTS: The optimum extraction process was adding 10 times amount of 75% ethanol, refluxing for 2 times, 1 h each time. CONCLUSION: This extraction process shows higher yield of total free organic acids in Pinellia ternate Breit and is available for industrial production. PMID- 15551971 TI - Dentistry on the road(map). PMID- 15551972 TI - Continuing education. PMID- 15551973 TI - Practical information. PMID- 15551974 TI - Insurance woes. PMID- 15551975 TI - Coverage dilemma. PMID- 15551976 TI - Flawed system. PMID- 15551977 TI - In vivo versus in vitro. PMID- 15551978 TI - Knowledge needed. PMID- 15551979 TI - Improving America's access to care: The National Institute of Dental and Craniofacial Research addresses oral health disparities. AB - BACKGROUND AND OVERVIEW: The National Institute of Dental and Craniofacial Research, or NIDCR, in 2001 sponsored the establishment of Centers for Research to Reduce Oral Health Disparities. The centers are based at Boston University; New York University; the University of Michigan; the University of Washington; and the University of California, San Francisco. Reflecting the importance of research to reduce disparities, the centers, along with related grants, represent one of the largest financial commitments ever made by the NIDCR. The centers are sponsored in part by the National Center on Minority Health and Health Disparities, or NCMHHD. Each of the five centers has forged partnerships that include ties with dental societies, state and local health agencies, community and migrant health centers, American Indian tribal nations and institutions that serve other diverse patient populations. CONCLUSIONS AND CLINICAL IMPLICATIONS: This network is attempting to address the needs of communities with poor oral health. A major part of the effort of these new centers is to build community networks and establish long-term relationships. Center investigators also recognize that solutions to these vexing problems must be built on an understanding of the social, economic, racial, educational, political and behavioral factors that affect most health care issues. PMID- 15551980 TI - Selectively preventing development of third molars in rats using electrosurgical energy. AB - BACKGROUND: Third molars are teeth with questionable value. People who never develop third molars avoid confronting the disease and pain these teeth often cause. Since third molars do not begin to develop until children are approximately five years of age, a window of opportunity exists to therapeutically prevent this tooth's development. The dentition of neonate rats possess developmental stages of molars similar to that of five-year-old children. This pilot study tests the hypothesis that third molars can be selectively prevented from developing. METHODS: Thirty-three neonate rats received a momentary pulse of electrosurgical energy to one of their maxillary tuberosities. The tuberosities on the contralateral sides received no treatment. Intraoral and radiographic examinations of sacrificed animals occurred when they were between 47 and 52 days old. Several tuberosity areas from sacrificed animals underwent histologic examination. RESULTS: Ten rats showed no intraoral or radiographic evidence of third molar development, and six developed smaller-than-normal third molars. Development of the maxilla also was affected frequently. One treated tuberosity area that was missing a third molar showed no histologic evidence of tooth-forming tissues, cyst formation or other significant abnormal tissue changes. CONCLUSIONS: Maxillary third molars can be selectively prevented from developing in rat pups at or near the time of tooth bud initiation; however, electrosurgical energy is too powerful and uncontrollable to reliably confine its damage to only the tooth-forming tissues. CLINICAL IMPLICATIONS: Third molars may be able to be selectively prevented from developing in growing children near the time of tooth bud initiation if less-differentiated precursor tooth-forming tissues can be selectively targeted. PMID- 15551981 TI - The effect of mechanical undercuts on the strength of composite repair. AB - BACKGROUND: The authors conducted an in vitro study to test the hypothesis that undercuts prepared in old composite restorations could improve the strength between the restoration and a flowable composite as repair material. METHODS: The authors used three composites to fabricate cylinders as repair substrates. The etched-only group was ground, etched, dried and built up with a flowable composite. For the undercut group, the authors introduced arrays of fissures on the surfaces before preparing the specimens for subsequent buildup in the manner described for the etched-only group. They made nonrepaired cylinders for baseline measures of strength. They sliced all finished cylinders into slender bars with a diamond saw. Flexure strength values were determined by a three-point-bending test. RESULTS: Nonrepaired bars exhibited statistically significantly higher flexure strength values than did repaired bars, as determined by Wilcoxon rank sum test. Two-way general linear model showed that both material (P < .0001) and undercut (P = .0207) exhibited a statistically significant influence on the repaired flexure strength. Repair substrate with elastic modulus close to that of repair material exhibited a greater percentage of recovery of the respective cohesive strength. Compared with the etched-only group, the undercut group yielded a higher mean flexure strength with one composite but a lower mean flexure strength with the other two. Examination of the fractured surfaces showed that a significant number of undercuts were filled only partially. CONCLUSION: Flexure strengths of repaired specimens always were lower than the cohesive flexure strengths of the materials being repaired. Undercuts did not generally improve repair strength. CLINICAL IMPLICATIONS: Small undercuts on the surface of composite often are difficult to fill completely, resulting in areas of stress concentration that result in no improvement in the repair strength. PMID- 15551982 TI - A systematic review of the performance of a laser fluorescence device for detecting caries. AB - BACKGROUND: The DIAGNOdent, or DD (KaVo America, Lake Zurich, Ill.), a device for detecting caries using laser fluorescence, has been growing in popularity during the past three years. Although several evaluations of its diagnostic performance have appeared in the literature, the range of reported performances is extensive. TYPES OF STUDIES REVIEWED: The authors conducted a systematic review of the literature to assess the diagnostic performance of the DD. Of 115 articles identified in the search, 25 studies were included in the review according to criteria requiring histologic validation and outcomes expressed as sensitivity and specificity values. RESULTS: For detection of dentinal caries, sensitivity values ranged widely (0.19 to 1.0), although most tended to be high. Specificity values exhibited a similar pattern, ranging from 0.52 to 1.0. In comparison with visual assessment methods, the DD exhibited a sensitivity value that was almost always higher and a specificity value that was almost always lower. The body of evidence is characterized largely by in vitro studies, so that generalization to the clinical setting is uncertain. Because caries thresholds varied substantially across the studies, synthesized estimates of performance were not possible. CONCLUSIONS AND CLINICAL IMPLICATIONS: The DD clearly is more sensitive than traditional diagnostic methods; however, the increased likelihood of false positive diagnoses compared with that with visual methods limits its usefulness as a principal diagnostic tool. PMID- 15551983 TI - Trigeminal neuralgia and glossopharyngeal neuralgia: two orofacial pain syndromes encountered by dentists. AB - BACKGROUND: Dentists frequently evaluate patients for oropharyngeal pain that may or may not eventually be related to oral pathology. Two rare neurological disorders that present with severe orofacial pain are trigeminal neuralgia, or TN, and glossopharyngeal neuralgia, or GPN. Both are secondary to cranial nerve compression by arteries and veins at the point at which the nerves exit the pons and brainstem. RESULTS: The authors present the results for two series of patients treated for TN and GPN. Significant success can be seen after intracranial microvascular decompression for both disorders, with low complication rates. Short- and long-term outcomes depend on proper patient selection. CLINICAL IMPLICATIONS: It is important for practitioners to recognize these syndromes and properly refer patients to a neurosurgeon experienced in treating such disorders. This can help the dentist and patient avoid oral procedures that will not alleviate the painful symptoms. PMID- 15551984 TI - An aid in sharpening explorers. PMID- 15551985 TI - Why switch to digital radiography? PMID- 15551986 TI - Medications' impact on oral health. AB - BACKGROUND: Over-the-counter and prescription drugs are used frequently, in large quantities and by many adults, particularly by those older than 65 years of age. A number of medications (prescription, over-the-counter, vitamins and minerals, herbal preparations) can affect oral health. With the population's aging, and as more drugs become available, dentists can expect to encounter medication-related oral side effects among their patients. TYPES OF STUDIES REVIEWED: The author reviewed studies that ranged from case reports to randomly controlled, double blinded studies. However, in view of the subject matter, the majority of findings are based on case reports. CONCLUSIONS: Since many patients regularly take medications, both prescribed and nonprescribed, dentists always must take a thorough medical history so that they can be aware of medication-related problems and the impact of medications on diagnosis and treatment planning. CLINICAL IMPLICATIONS: Dentists must be aware of the potential oral tissue complications that medications can create and develop appropriate treatment plans for their patients that consider the oral health impact of the medications they take. PMID- 15551987 TI - The standard of care in dentistry: Where did it come from? How has it evolved? AB - BACKGROUND: The author explores the topic of the standard of care. He reviews the initial application of the standard to health care professionals, as well as its evolution from an 1898 legal case in which 10 elements of the standard of care were stated. CONCLUSIONS: The standard of care continually evolves with the advent of new materials, new procedures and new court rulings. Before applying the standard of care, dentists should consider new available treatments, as well as their state's current interpretation of the standard of care. PRACTICE IMPLICATIONS: The standard of care should be applied to all dentists when patients claim alleged malpractice. It should be tempered by changes in the practice of dentistry, which dentists should be aware of. PMID- 15551988 TI - Maximizing the physical use of the office. AB - When referring to the physical plant in a dental practice, I am referring mainly to the use of dental chairs, because this is where dental practice production occurs. By maximizing the use of chairs or adding the necessary number of chairs to a practice, any office can grow and experience a proper patient flow. Since most offices have significantly high rates of no-shows, last-minute cancellations and overdue patients, the potential for growth is enormous. By using the schedule and chairs properly, the office has the opportunity to maximize production, create convenient appointments for patients and significantly increase practice profitability. PMID- 15551989 TI - Self-reported satisfaction of enrollees in capitated and fee-for-service dental benefit plans. AB - BACKGROUND: This article examines the impact of capitated, or CAP, and fee-for service, or FFS, dental benefit plans on the enrollees' satisfaction with their plans and their satisfaction with their dentists. METHODS: The authors selected four dental markets: California, New Jersey, Michigan and North Carolina. Eight Fortune 500 companies participated. Enrollees were selected randomly and interviewed about their experiences with their dental plans. The sample consisted of 2,340 respondents, of whom 42.3 percent were enrolled in CAP plans and 57.7 percent in FFS plans. RESULTS: The major findings were that those enrolled in FFS plans were four times more likely to be very satisfied than dissatisfied with their dental plans than were those in CAP plans. The FFS plan enrollees were 16 times more likely to be very satisfied than dissatisfied with their dentists than were those in CAP plans. CONCLUSION: Enrollees generally were satisfied with their plans and their dentists but those in FFS plans were the most satisfied. The higher the premium paid, the higher the level of satisfaction. PRACTICE IMPLICATIONS: Enrollees with perceived unmet needs were less satisfied with their dental benefit plans and dentists. Taking care of needs is the most significant thing dentists can do to affect patients' satisfaction. PMID- 15551990 TI - Curing performance of a new-generation light-emitting diode dental curing unit. AB - BACKGROUND; Recent technological advances have resulted in the marketing of high powered, or HP, battery-operated light-emitting diode, or LED, dental curing lights. The authors examine the curing efficiency and peak polymerization temperature, or Tp, of a new HP LED curing light. METHODS: The authors studied four visible light-curing, or VLC, units: HP LED (A), first-generation LED (B), conventional halogen (C) and high-intensity halogen (D). They determined the depth of cure, or DOC; adhesion; and Tp of three types of VLC resin-based composites after exposure to each light. The exposure times for units A and D were one-half those for units B and C. RESULTS: The power density of unit A was 1,000 milliwatts per square centimeter, which was comparable to that of unit D with turbo charge. The DOC and adhesion attained for all three resin-based composites after being light cured by unit A for a 10-second exposure time were equivalent to those after being light cured by unit D for a 10-second exposure time and to those after being light cured by units B and C for 20-second exposure times. The resin-based composites light cured by unit A attained significantly lower Tps than did those light cured by unit D at equivalent cure, or exposure, times and by unit C at twice the cure time. CONCLUSIONS: The authors found that Unit A effectively cured the resin-based composites at one-half the cure time of units B and C and at the same time as unit D, while maintaining low Tp. CLINICAL IMPLICATIONS: The battery-operated HP LED curing light might be an effective, time-saving alternative for clinicians to use in light curing resin-based composites. PMID- 15551991 TI - High court rules ERISA pre-empts negligence claims against HMOs. PMID- 15551992 TI - For the dental patient. Missing a tooth? A single-tooth implant may be for you. PMID- 15551993 TI - [Biosafety in STD/AIDS: conditioning factors of nursing workers' adherence to precaution measures]. AB - Aiming to identify the conditioning factors of nursing workers' adherence to precaution/isolation in caregiving, this descriptive study was conducted with the target population consisted of nurses representing medium-sized and large hospitals in the city of Sao Paulo. Of the 79 institutions that met the established criteria, 15 (18.98%) comprised the sample of the study. In a collective meeting, the focal group technique was used for data collection. After obtaining the participants' consent, discussions were recorded on cassette tapes and then transcribed. Data was organized as proposed by Bardin (1977), thematic units were extracted and two empirical analysis categories were defined, which were termed institutional and individual conditioners. PMID- 15551994 TI - [Compliance with the treatment of chronic pain and health control locus]. AB - The objective was to assess compliance of patients with nonl-oncologic pain and identify relationships between beliefs of health control and compliance. Thirty patients were evaluated over a period of six months, in five interviews. Compliance was classified as total, partial and non-compliance, and the Index of Correct Medication Ingestion (ICMI) was calculated. The belief about health control was evaluated by the Multidimensional Health Locus of Control Scale. High rate of partial compliance and non-compliance with the treatment (40.0% to 56.7%), ICMI means between 57.2% and 69.5% were observed, and that patients with higher "internality" beliefs were less compliant with the treatment. PMID- 15551995 TI - [Social support in the rehabilitation of mastectomized women: the role of the sexual partner]. AB - This study identified the types of social support offered by the sexual partners of women with breast cancer and their perceptions about this support. Nine partners of women in this condition participated in the study. Data was collected through interviews and submitted to content analysis. Partners perceived themselves as important elements of social support to their wives, offering affection, stimulus to self-care and help with household chores. Difficulties faced in offering social support involved sexual relations, communication channels, the feeling of impotence and insecurity to deal with the implications of the diagnosis and to reorganize household chores. PMID- 15551996 TI - [King's theory and its interface with the "Family Health" Program]. AB - This study of epistemological intention is about King's goal achievement theory and its contributions on knowledge and practicing for nurses who work in the "Family Health" Program (FHP), Brazil. The analysis component, based on the Meleis' Method of Theory Analysis concerning critical thought on the theory, was applied as well as reading and thoughts on FHP guidelines and proposals in order to verify the interface between the two paradigms in the study. The outcome of the study proves that the two approaching proposals are coherent. Such coherence drives authors to recommend the adoption of King's teachings concerning FHP practice for nursing to obtain greater and better results in its actions. PMID- 15551997 TI - [The process of organizational culture formation in a philanthropic hospital]. AB - This study was carried out in a philanthropic medium-size hospital institution in Sao Paulo - Brazil, aiming to disclose the cultural features of the institution. The adopted methodology was the qualitative study, following the steps proposed by Thevenet: document analysis, interview and observation. The analysis showed that when a new professional group starts working in an institution, it considers that some values must be changed. This change means to restructure the management of the organization and the people involved in it, facing the conflict posed by changing or preserving the old system. PMID- 15551998 TI - [The meaning of team work in the rehabilitation of people with congenital craniofacial malformation]. AB - This study aimed to understand the meaning of teamwork of rehabilitation professionals in craniofacial anomalie. It was carried out a phenomenological analysis for contemplating the understanding and interpretation of the sense considering the subject. Twelve professionals from different areas were interviewed, guided to the subject: What does work in a team on the rehabilitation of craniofacial anomalies mean to you? These themes were brought up: training for the task, difficulty in working with a team, relationship with patient and family, work conditions and the professional insertion in the team. The analysis sought to reflect the phenomenon engendering convergences and divergences to express the differences and continuous learning. PMID- 15551999 TI - [Hospitalized children' perceptions on exams]. AB - The study had as objective to identify the hospitalized children's perception, in school age, about the accomplishment of exams. The research of descriptive nature was used, prevailing the investigation of qualitative character. The results revealed that the children cooperate with exams despite the fear. The fear was related to the dread of the physical pain, being hurt, or even the unknown. It is considered that the child notes the exam accomplishment as part of his/ her treatment and he/she would like to receive explanations about the procedure. PMID- 15552000 TI - [Nursing diagnosis of patients in the preoperatory period of cardiac surgery]. AB - The purpose of this study was to identify nursing diagnosis of patients in the preoperative period of cardiac surgery. One instrument was elaborated and validated for data collection. Such instrument was organized according to Horta's Conceptual Model. Seventeen adult patients in the preoperative period of cardiac surgery were evaluated, of whom 09 were males and 08 were females with the average of 58 years old. After data collection, the nursing diagnoses were identified based on NANDA's Taxonomy I. The nursing diagnoses identified with a higher frequency than 50% were: Activity Intolerance (88.2%), Infection risk (76.5%), Risk for peripheral neurovascular dysfunction (76.5%), Knowledge deficit (on the disease, surgery and anesthesia) (76.%), Altered cardiopulmonary tissue perfusion (70.6%), Ineffective airway pattern (70.6%); Pain (70.6%), Altered Sexual Patterns (58.8%) and Sleep Pattern Disturbance (52.9%). It was observed a predominance of diagnoses related to physiological needs. PMID- 15552001 TI - [Cost teaching in graduate nursing schools]. AB - The present study intends to analyse the implement of content related to Costs in the Management Applied to Nursing subject in Graduate Nursing Schools in Sao Paulo State. The descriptive, exploratory associative study with a quantitative approach was put into effect in 28 professionals at the Nursing Schools. The results demonstrated that 19 (67.86%) professors teach that subject in graduation. No statistically significant correlation or association has been found among Cost Teaching, the Number of Post Graduation staff and the type of Institutions they work for (Public or Private). PMID- 15552002 TI - [Training and development of the nursing staff: a model of spread sheet cost]. AB - This paper presents a model of spread sheet cost for training and development programs to the nursing staff in a hospital organization. Significant items of the total cost have been considered in relation to its elaboration and proper functioning. This model is divided into four parts: Item 1--data related to the training programs; Item 2--direct costs of these programs; Item 3--indirect costs (Continuum Educational Center structure), and Item 4--cost amount. The use of spread sheet cost may provide knowledge and managementfor the nurses and nurse managers. However, the related costs should be reviewed according to each service. PMID- 15552003 TI - [Holism only in theory: the struggle of nursing students feelings about their education process]. AB - Despite the speech of the human being's integration, we have observed that the nurse professional formation emphasizes the technical dimension, leaving out the professional's internal growth. Searching for understanding that process, our goal is to identify and to analyze the perception and feelings of nursing teaching staff, related to their formation as a person and as a professional in the of Nursing scope. The reports that were submitted to the content analysis pointed out two categories: Holism only in theory and Scheme of the feelings, revealing a stage of many discoveries and little space for the student person. The results of the research offered important elements to think about it by the Schools, visualizing the student's formation as an integral and integrated person. PMID- 15552004 TI - [Weaning experiences among women from the same family]. AB - This work is a case study that aimed to acknowledge the reasons that persuade women-mothers from a same family to wean their children, and identify the feelings that accompany these women during this process. A semi-structured interview was used for data collection, and the analysis was done using Minayo's (1993) methodological orientations. It was concluded that weaning takes place due to various reasons such as: a new pregnancy, child's age, mother's or child's disease, and breast refusal by the child. It was verified that weaning can be soothing for some women while for others it is accompanied by suffering. Through the studied women-mother's speeches, it was observed that policymakers' discourses on breast-feeding seem to influence a lot on this practice, having better results on the youngest ones than the previous generations of the family. PMID- 15552005 TI - [Epistemology of psychiatric and mental health nursing: the need to develop skills in nursing training]. PMID- 15552006 TI - [Cooperation between a surgeon and an interventional radiologist in cases of internal bleeding]. AB - INTRODUCTION: Internal bleeding still remains a serious condition, which must be urgently diagnosed and treated. In the Faculty Hospital in Hradec Kralove the diagnosis of the internal bleeding source and its consequent treatment is managed under cooperation between a surgeon and an interventional radiologist. A CASE REVIEW: In the case-review section of this report, a total number of five patients with serious cases of internal bleeding, the source of which was diagnosed and treated under cooperation between the above mentioned specialists is presented. The patients concerned suffered from the following: a trauma to the liver parenchyma, posttraumatic arterioportal shunts, an intraheparic aneurysm, a relaps of the uterine carcinoma with hemorrhaging into the colon and the vagina, bleeding into retroperitoneum. In all the cases the lesion was successfully treated and the hemorrhage was managed. DISCUSSION: Exact location of the bleeding artery during an arteriogragraphic examination is a prerequisite of a successful treatment of the condition. Both, the absorbable materials which allow gradual recanalization of the embolized vessel, and the non-absorbable materials may be used to conduct embolization of the affected vessel. It is necessary to haemodynamically stabilize the patient prior to the procedure. CONCLUSION: In many cases, the cooperation between a surgeon and an interventional radiologist allows for the source of the internal bleeding to be located and managed. It is indicated in certain cases of benign and malignant disorders and in some cases of the parenchymatose organs trauma. PMID- 15552007 TI - [Unusual cause of the acute appendicitis]. AB - Exogenous foreign particles in the appendix rarely case appendicitis. Their prevalence is reported to be 0.0005%. This fact makes both lay and medical community underestimate possible complications related to the presence of foreign particles in the appendix lumen. Our case review documents risks related with swallowing a sharp foreign particle. The particle concerned was a tooth root--on prosthesis with an acrylic crown, which caused acute appendicitis with the appendical wall perforation by the above mentioned foreign body. PMID- 15552008 TI - [Cryolization in the inoperable breast carcinoma treatment]. AB - The authors present a case review describing a method of cryolization. They employed this method in the case of a patient with an extensive advanced breast carcinoma with multiple metastases located in the thoracic wall, in order to make the patient's state surgically manageable. PMID- 15552009 TI - [Secondary malignant tumors of the mammary gland]. AB - The authors present three case-reviews of rare metastatic affections of the female mammary gland by the following disorders--a malignant lymphoma, a Grawitz carcinoma of the kidney and by a malignant melanoma. Histological examinations of the tumors and restrospective examinations of the respective patients anamneses resulted in the primary focus determination in two cases. PMID- 15552010 TI - [A contribution to contemporary absolute and relative contraindications for breast conserving surgeries (BCS) in early carcinoma of the breast in the first or second stage]. AB - Absolute and relative contraindications for BCS (Breast Conserving Surgery) are stated according to the EUSOMA (European Society of Mastology). Statement includes the risk factors for local recidive, and unfavourable progress of the illness itself. Stress is put on the health treatment fallowing BCS. PMID- 15552011 TI - [A contemporary approach to ductal and lobular carcinoma of the mammary gland in situ]. AB - The statement discusses epidemiology data about DCIS and LCIS, their classification evaluates predictive prognosis factors. Surgical methods are stated, as well as the following health treatment. We present the basic oncology reguirement for the surgical treatment at DCIS, meaning "tumor free margins". DCIS is clinically more serious and significant than LCIS. PMID- 15552012 TI - [Mesenchymal extraperitoneal tumors in the elderly]. AB - Incidence rates of solid tumors increase with age in our population. Epithelial tumors are the most frequent tumors of the old age. On the contrary, extraperitoneal mesenchymal tumors are rare. Surgical treatment remains the most important and the most effective therapeutic modality to manage solid tumors. The risk level of the surgical treatment in the elderly is influenced by many factors, which may often be substancially reduced. Therefore, the elderly should not be excluded from curative surgical procedures on their calendary age basis. PMID- 15552013 TI - [Laser palliative destruction of rectum carcinoma--case reports]. AB - The palliative treatment of developed primary or recurrent rectum carcinoma is still an important issue in treatment of this disease. The aim of the surgery is in such case to remove the tumorous obstacle and to prevent the development of ileus, to reduce pain, to limit the volume of tumorous tissue, to prevent bleeding and possible incontinence or mucous secretion. The transanal laser thermodestruction of the tumor represents one of the possible ways of palliative treatment of rectum carcinoma. The method is very considerate even for the high risk patients. On the other hand it requires a short-term hospitalization. The method of transanal laser evaporisation of carcinoma is also important from the ethical point of view: in many cases it may prevent the creation of palliative derivative colostomy. This fact is very important especially in patients with the terminal stage of the malign carcinoma because they live the rest of their lives in acceptable conditions. The important benefits are described in three case histories. The method is presented as an urgent surgery in solution of ileus which is based on tumorous rectum stenosis. PMID- 15552014 TI - [War surgery at the turn of the century. Some aspects of surgeon's education and preparation]. AB - Authors describe current situation in surgical residents training in the military environment with focus on military surgery and surgery. The recent changes running in Czech Army Military Medical Service and changes associated with the new law regulating postgradual education in medical subjects require quite new approach to residents training in Czech Republic. This process will have to set up new educational goals, curricula remake and method of teaching quality measurement. PMID- 15552015 TI - [Limited open dissection of the flexor retinaculum in the surgical treatment of carpal tunnel syndrome]. AB - Decompression of the median nerve with flexor retinaculum release is the keystone of the surgical treatment of the carpal tunnel syndrome. There is still a scientific discussion about the risks and benefits of the conventional surgical techniques (radical open versus endoscopic dissection). In our mind limited-open release of the flexor retinaculum seems to be an optimal method, because of the fact that combines the predominant benefits of the conventional techniques (e.g. direct visual exploration of the nerve, decrease of the postoperative pain and morbidity, shortening of the postoperative reconvalescence etc.). Thirty-eight patients with carpal tunnel syndrome treated with an open-limited release were followed up. Any wound infection or other healing complication, any chronic pain of the palm (as a result of a palmar cutaneous branch of the median nerve injury), or painful scarring were remarked postoperatively. One patient (2.6%) underwent reoperation because of the symptoms recurrence. Other patients with complete resolution of the symptomatology were satisfied with the surgery. PMID- 15552016 TI - [Conversion of an external fixation to that with an intramedullary pin in cases of complicated diaphyseal fractures]. AB - In this study the authors present a method of conversion of external fixations to closed intramedullary ones in the treatment of complicated diaphyseal fractures of the calf and femur. According to the conditions, the fixation is secured by a pin. The above mentioned procedure is advantageous especially in cases of patients with open fractures, in patients with polytraumas as well as in patients with a threatening or with a present compartment syndrome. Treatment of the above fractures is not always simple and represents a serious problem in everyday traumatological practice. Treatment of fractures using the above procedure has very good results with a minimum of complications. Although, nowadays, the skeletal defect can be solved and therefore the bearing capacity of the extremity be renewed, damages to the muscles are often irreparable and cause a permanent invalidity. PMID- 15552017 TI - [Role of videomediastinoscopy in diagnostic procedures of pathological conditions in the mediastinum]. AB - Due to recent advances in videotechnology and a worldwide comback of invasive staging methods of the lung cancer, mediastinoscopy has been revived. In the Surgical Clinic of the Faculty Hospital in Plzen, the authors carried out 54 videomagnetoscopic examinations of 53 patients aged 58.1 years on average, in total, during the period from 2000 till 2003. Mostly, in 41 cases (76%) the examination was performed as a part of the lung cancer staging procedure. None of the patients exited in direct correlation with the surgical procedure. Complications, not requiring reoperation, were reported in 2 cases. The lung carcinoma staging findings agreed with the postoperative N disease classification in 86% of the examined cases. Five cases (14%) were underestimated and always the superior lobe carcinomas were concerned, out of which four of them were on the left. At the same time, the non-invasive staging examination using computer tomography (CT) in the mediastinal lung cancer dissemination examination, proved to show a high rate of false-positive findings, when compared to that with videomediastinoscopy. Mediastinoscopy remains the supreme method in determining other pathological affections of the middle mediastinum, if properly indicated. The authors carry out the above miniinvasive examination as a part of the lung cancer staging method in all patients with the CT-confirmed mediastinal lymphonodes enlarged over 1 cm. They consider this method a necessary part of this disease staging protocol, in cases of the left superior lung lobe tumors combined with videothoracoscopy. At the same time the authors recommend it as the method of choice in diagnosing other pathological processes of the middle mediastinum, mainly for its high efficiency rate and its low rates of complications. PMID- 15552018 TI - [Thyroid gland surgery--a contribution to the discussion on its surgical technique]. AB - The authors present their thyroid gland operation results from the point of view of the current discussion on various techniques of the thyroid surgical procedures. PMID- 15552019 TI - [Patients' perioperative worries and experiences]. AB - AIM: The aim of this study was to detect the main causes of patients' preoperative distress and their subjective experiences. METHODS: After institutional ethic committee approval and patients consent an anonymous questionnaire was randomly distributed on the first and second postoperative day. Its content were basic demography, preoperative anxiety level using five point verbal scale, main cause; of anxiety, the worst postoperative experience, postoperative pain intensity (11 point scale) and its treatment incidence of postoperative nausea and vomiting (PONV) and other side effects of the surgery and the anesthesia. Correlations with specific patients' characteristics were searched for to determine the risk groups. RESULTS: Two hundred filled in questionnaires out of the 244 distributed could be used for further evaluation. Sixty-two per cent of patients suffered from preoperative anxiety, more women (p < 0.01) and more frequently after intra-abdominal (intra-thoracic) surgeries (p < 0.05). Forty-three per cent of patients had no subjective problems following surgery, others suffered more frequently from the postoperative pain (18.5 percent), PONV (11.5 percent) and postoperative procedures, e.g. changing of dressing, drains and from the postoperative regime. There was a correlation between PONV and both the intensity of postoperative pain and anamnesis of PONV during previous surgery (p < 0.01). PMID- 15552020 TI - [Genetics of inflammatory bowel disease]. AB - The Inflammatory Bowel Disease (IBD) are multifactorial diseases involving the interaction of genetic and environmental factors. In genetic terms, the IBD are polygenic and multigenic disorders with incomplete penetrance. In the late decade, investigators have applied the complementary techniques of genome-wide scanning and candidate gene analysis to search susceptibility genes. The IBD susceptibility regions, widely replicated, are in chromosomes 16 (IBD1), 12 (IBD2) and 6 (IBD3). Recently, a significant association have been reported with Crohn's disease and NOD2/ CARD15 gene. This gene is an appropriate candidate gene because its localization and function. More studies is necessary to confirm this association, search an other variants of this gene and other candidate gene. This studies provide best comprehension of the disease pathogenesis and deliver clinical application. PMID- 15552021 TI - [Rheumatologic manifestations of parvovirus B19 infection]. AB - Rheumatologic manifestations of parvovirus B19 infection is a recent individualization. They are rather frequent and varied and meet at the young adult's. Generally, it is responsible for acute, bilateral and symmetrical arthritis, usually involving distal joints and sometimes associated to signs caused by viral infections. Prolonged articular forms were discribed but are rare being able to sometimes feign a nocive rheumatoid arthritis because of their clinical aspect and of a seropositivity. Axial manifestations were also reported. The diagnosis of the recent infection by the parvovirus B19 can be confirmed by ELISA identification of specific IgM antibodies. The mechanism of the articular manifestations is still unknown and the link between parvovirus B19 and systemic vasculitis is questioned. PMID- 15552022 TI - [Evaluation by Doppler echocardiography of hemodynamic profil of mitral bileaflet prothesis. Report on 90 cases]. AB - From 1981 to 1999 Mitral bileaflet prosthesis was implanted to 90 patients. Doppler echocardiography was performed for these patients between January and March 2002 with a mean deadline of 111 months after the intervention. 36 were women (40%) and 54 were men (60%) the mean age was 41 years (20 - 70 years) The mitral bileaflet prosthesis was a Saint Jude in 65 cases, Jyros 8 cases, Carbomedics 7 cases, Sorin Bicarbon 7 cases, Edwards Duromedics 2 cases et an ON X in one case. The maximal transprosthetic gradient was 15.7 mm Hg +/- 5.06 (6.25 mm Hg). The mean transprosthetic gradient was 5.6 mm Hg +/- 1.07 (39.5 mmHg). The mean prosthesis functional area 2.37 cm2 +/- 0.44 (1.75 cm2 et 3.60 cm2). Maximal gradient, mean gradient and prosthesis functional area are independent from kind mitral bileaflet prosthesis and from the prosthesis size. PMID- 15552023 TI - [The delivery of macrosomic infants weighing 4500 g and more. A report of 61 cases]. AB - The objective of this study is to review the difficulties in prediction of great macrosomia, to assess trial of labor results, and to confirm the increased risk of perinatal complications. MATERIAL AND METHOD: in this retrospective study we analyzed 61 deliveries of infants with birth weights > 4500 g in one year period. This group was compared with a group of infants weighing between 4000 and 4500 g (339 cases) born during the same period. Elective cesarean delivery was performed for 3 cases of the second group because of overestimation of fetal weight. 38 cases of the first group (62.3%) were delivered vaginally after underestimation of fetal weight. Incidence of shoulder dystocia in vaginal delivery was 13.5% in the first group and 5.32% in the second one, but difference was not statistically significant (p = 0.07). Difference between incidence of hypoglycemia in the first group (18.03%) and in the second one (2.06%) was highly significant (p = 0.00006) There is no reliable method for prediction of fetal weight > than 4500 g. With literature review, we confirm the increased risk of shoulder dystocia, birth asphyxia and hypoglycemia for these infants. So, we believe that cesarean delivery is justified in all cases of fetal weight estimation > 4500g. PMID- 15552024 TI - [Epilepsy and pregnancy: necessity of a combined care]. AB - Pregnant women with epilepsy are at risk for a variety of complications, that require an interdisciplinary cooperation between neurologists, pediatricians and obstetricians. We report a retrospective study of 50 pregnant women with epilepsy over a 5 year period. Phenobarbital is largely the most used (74%). During pregnancy, an increase in seizure frequency was noticed in 27 cases (54%). Baribituric were essentially maintained in 74% of cases. We have reported a relatively high intervention rate during the delivery. Forty eight babies (96%) were born at term. There were no cases of neonatal haemorrhage or perinatal death. Symptoms of drug impregnation were found in 7 cases (14%). Four children born to epileptic mothers treated during pregnancy had congenital malformations. Pregnancy doesn't have a catastrophic effect on the course of epilepsiy apart from the risk of an increase in the frequency of the fits. This risk must be reduced thanks to an optimal use of an anti-convulsive therapy before and during pregnancy. The major risk being foetal, mainly foetal malformations and hemorrhages. PMID- 15552025 TI - [Carcinoid tumors of the appendix. A report of 46 cases]. AB - Carcinoid tumors of the appendix are rare. The appendix constitutes the most frequent localization of these tumors. We studied of a retrospective way all cases that had an appendectomy in the service of general surgery of Sfax on a period of 7 years going from 1995 to 2001. During this period, 9584 patients had an appendectomy in our service. Among which, 46 cases (0.48%) of carcinoid tumor of the appendix have been diagnosed. In more than 95% of cases, patients were admitted due to acute appendicitis; but none of them had the signs of the carcinoid syndrome. The treatment was a simple appendectomy in 44 cases (95%). The right hemicolectomy was necessary in two cases, because of an invasion of the appendicular base in a case and a tumor greater than 2 cm in the other case. There is no reported case of lymph node or systemic metastasis. The authors discuss data in the literature and analyse the features of the clinical picture, diagnosis, and try to define a convenient attitude of treatment of these tumors. PMID- 15552026 TI - [Nonresponse to hepatits B vaccine in health care workers]. AB - The authors report the results of an investigation of witness cases realised in collaboration between Occupational Medecine Service and Immunology Laboratory of Hedi CHAKER University Hospital SFAX during the year 2000. The purpose was to search the genetic control of the HLA class I system for the non-response to hepatitis B vaccine and to evaluate the contribution of other favorite factors as tabac, sex, age. Thus, in a population of 32 healthy agents found nonresponders to hepatitis B vaccine by the titers of anti HBs antibody, we have performed the HLA-A, -B phenotypes by the technique of complement dependent microcytotoxicity. The frequency of studied HLA class I antigens, was compared for the non responders group, to the frequency observed in witnesses group, done with 52 responders healthy agents. The rate of nonresponsiveness hepatitis B vaccine was evaluated up to 5 %. Statistically significant difference was observed for HLA A1 and-B44 markers showing fraquencies which were considerably higher in the non responders than in witnesses. PMID- 15552027 TI - [Pustular psoriasis in childhood in 15 cases]. AB - Psoriasis is a common, chronic and recurrent, inflammatory disease of the skin. Children were affected in 30% of cases. Severe form can be observed: pustular psoriasis (PP), psoriatic arthropathy and erythrodermic psoriasis. There were 15 children with PP, with an age range of 1-15.5 years, (mean age 7.56 years). Psoriasis constituted 5% of the total dermatological disorders in children. The severe forms of psoriasis were rare in children Annular PP was the most common form of PP in children. Juvenile PP had generally better prognostic than in adults, but the evolution is characterized by recurrences. PMID- 15552028 TI - [Toxocariasis: under-estimated disease. Report of 9 cases]. AB - Toxocariasis or visceral larva migrans is a parasitosis due to the migration in the human organism of animal ascarid larvae. Its importance is under-estimated and the reported tunisian cases are rare. We report nine cases of toxocariasis, noted between January of 2000 to March of 2002, in the laboratory of parasitology mycology in La Rabta Hospital. They were six children and three adults. The clinical forms are varied: ophthalmologic form (5 cases), general syndrome (1 case), oedema (1 case) and hypereosinophilia (2 cases), and confirmed serologically by the presence of anti Toxocara canis antibody using ELISA test. Visceral larva migrans should be kept in mind to avoid severe forms such as ophthalmologic forms. PMID- 15552029 TI - [Potter's reno-facial syndrome]. AB - Potter's reno-facial syndrome is a rare innate abnormality. We bring 4 observations repertoried at the maternity of military hospital of Tunis over a period of 6 years (1997 - 2002). The purpose of our work is to determine after a review of the literature the echographic and foetopathologic characteristics, and the forecast of this syndrome. The frequency of the bilateral renale agenesis is of 0.27 per thousand. Positive diagnosis bases essentially on the ultrasound of the 2th, or the 3-th trimester. The signs of appeal are essentially the oligoamnios associated to an hypotrophy. The caryotype is systematic to eliminate an associeted chromosomic abnormality. Foetopathologic exam is usefull for the diagnosis. Main abnormality except the urinary pathology is the lung hypoplasia. Therapeutic interruption of the pregnancy in this situation not compatible with the extra-uterine life., only type IV authorize the development of the pregnancy according to echographic data and of foetal urinaire biochemistry. We insist on the early practice of the morphological ultrasound between 20 - 22 weeks for the diagnosis of foetal abnormalities and the place of the genetic advice in association with the geneticist in the coverage of the couple. PMID- 15552030 TI - [Hypertrophic osteoarthropathy revealing a malignant thymoma in a child]. AB - Association of hypertrophic osteoarthropathy and malignant thymoma is very rare in children. Authors report a case of a 14 year old girl followed for rheumatic fever since she was 8 years old. She was explored for arthralgia. The exam showed hippocratic fingers, soft tissues hypertrophy of lower limbs and dyspnoea. Plain radiography had demonstrated symmetrical ossifying periostisis in tubular bones of lower limbs evocating hypertrophic osteoarthropathy diagnosis. Chest radiography has demonstrated bilobate anterior mediastinal mass. Ultrasonography and MRI revealed a voluminous anterior mediastinal mass with cystic and fleshy components. Thymic adenocarcinoma diagnosis was made by surgical biopsy. PMID- 15552031 TI - [Malign Brenner's tumor particularities and coverage]. AB - Brenner's tumour of the ovary is rare, mostly benign. It was described for the first time by Mac-Naughton-Jones in 1898. Von Numers listed the first malign form in 1945 that represents 2 % of tumours. On the basis of a personal report and of 160 cases listed in the literature, we propose to study anatomoclinical and epidemiological elements of this hurt and to study histological particularities and various therapeutic processes. Malignant Brenner's tumour arises mostly in peri and postmenopausal period. Clinical signs are not specific. Histological exam puts the problem of the choice of the histological malignant criteria. Metastasis occurs in half of cases. They are mostly loco-regional. The treatment is essentially surgical. The efficiency of an additive treatment seems to be debatable. PMID- 15552032 TI - [Duodenal varix rupture a rare cause of gastrointestinal bleeding from portal hypertension]. AB - The duodenal varix rupture is a rare and serious complication of portal hypertension. The authors report one case of duodenal varix rupture in a 38 year old woman, revealing portal hypertension, which caused a massive gastro intestinal bleeding. The endoscopy didn't allow to conclude to a diagnosis. An emergent laparotomy was necessary regarding the massive bleeding. A duodenal varix suture ligation was performed after pyloroduodenotomy. She had a well recovery. After a review of literature, the authors conclude that this abnormal situation is rare, its diagnostic is difficult and it has several therapeutic strategies. PMID- 15552033 TI - [Doctor Mahmoud el Materi one of the first physicians and militants of Tunisia]. AB - Mahmoud el Materi was undoubtedly, one of the main leading figures of contemporary tunisian history. Open and brillant, he detained a vast knowledge, but was nevertheless a man of moderation and humility, and a humanist who dedicated his entire professional and militating life to the service of the people of Tunisia. In a country, stricken then by poverty and widespread human misery, he was a remarkable militant who fought for independence and greatly contributed to the rebirth of modern Tunisia. PMID- 15552034 TI - [Genetic structure of Tujia as revealed by Y chromosomes]. AB - Tujia people call themselves "Bizika", which means aboriginal. Genetic study of Tujia is virtually absent. To characterize the genetic structure of Tujia,the distribution of 14 Y haplogroups was studied in Tujia populations sampled from Enshi, Hubei (31 males) and Jishou, Hunan (68 males). A total of eight haplogroups were observed in the Enshi and Jishou populations. The haplogroup frequencies of Tujia were compared with the frequencies of other related ethnic groups, including Northern Han, Southern Han, Tibetan-Burman speaking populations,Daic and Hmong-Mien. The principal component (PC) analysis was conducted and the PCs were plotted to explore the historical migrations. In addition, partial correlation analysis was performed to study the relationship between the first three PCs and the haplogroups. The PC2 revealed a cluster of Tujia groups including Longshan,Yongshun, and Enshi with Lahu, suggesting possible interaction between Tujia and the Di-Qiang groups. However,a similarity between Han and Tujia populations, though differentiated, were also observed. We postulated, by incorporating the results of archaeological and historical evidences, that the Ba people, the ancestors of the Tujia,might be related with Di-Qiang groups and inhabited the Tujia area initially before a substantial interaction with Han and other ethnic groups. PMID- 15552035 TI - [Principal component analysis of Y-chromosome haplotype distribution in 18 ethnic groups in Yunnan Province]. AB - Based on the historical records, 18 of the 26 ethnic groups in Yunnan Province are the descendant populations of three ancient tribes, Bai-Yue, Bai-Pu and Di Qiang, linguistically belonging to the Daic, Austro-Asiatic and Tibeto-Burman, respectively. In order to trace the origins of these native ethnic groups, a total of 13 East Asian specific Y-chromosome biallelic markers were used to study the genetic structure of 20 local populations covering all the 18 ethnic groups in Yunnan Province. Haplotypes were analysis by PCR-RFLP method. Our results showed that H11 and H12 were the predominant haplotypes in the descendant populations of Bai-Yue tribe. H5, H6 and H8 were the dominant haplotypes in Di Qiang descendants, and the frequencies of H6, H8 and H11 were very high in the descendant populations of Bai-Pu. To investigate relationships among 20 populations, a three dimensional PC analysis were performed based on the distribution of the 13 haplotypes. All populations were divided into two clusters in the PC plot. The first cluster was mainly composed by the descendant populations of Bai-Yue, and the second one was mainly composed by the descendants of Di-Qiang tribe. This result indicated that Bai-Yue and Di-Qiang's paternal lineage had different origins, which was in agreement with the historical documents and linguistic classification. PMID- 15552036 TI - [Regulation of pre-mRNA alternative spicing in U251 cells by TPA]. AB - By using the mini-gene construct containing partial sequence of Bcl-X gene as model, we examined the function of TPA on Bcl-X pre-mRNA alternative splicing in vivo and vitro with RT-PCR and site-directed mutagesis assay. The results show that PKA signaling system can regulate Bcl-X pre-mRNA alternative splicing, the possible mechanism is that the responsible sequence affect the choice of the 5' downstream or upstream splice site of Bcl-X pre-mRNA. PMID- 15552037 TI - [Detecting genetic polymorphisms of CYP1 A1 and GSTM1 simultaneously with oligonucleotide microarray]. AB - Cytochrome P4501A1 plays a major role in the bioactivation of a number of tobacco procarcinogens. Glutathione S-transferase( GSTM1), a member of the class of GST gene family, has been shown to be polymorphic because of gene deletion resulting in a failure to express the GSTM1 gene in 50% approximately 60% of individuals. Some CYP1 A1/GSTM1 null genotype combinations seem to predispose the lung, esophagus, and oral cavity of smokers to an even higher risk for cancer or DNA damage, requiring, however, confirmation. An easy and reliable oligonuleotide microarray approach validated through direct sequencing method is developed in order to accurately detect single nucleotide polymorphisms of CYP1 A1 gene and discriminate the presence and absence of GSTM1 gene. The m1 (Msp I) and m2 (Ile462Val) polymorphisms of CYP1 A1 gene and GSTM1 null genotype were also determined in a random population of 84 healthy, unrelated volunteers with developed microarray-based method. Of 84 cases, 47.6% were calssified as GSTM1 null, close to the published data. It's interesting that there lack three genotypes of m1 -m2 locus in the population: TT-AG, TT-GG and TC-GG. However, according to the data of the genotype frequencies independently happened at both m1 and m2 site, the combination frequencies of above three genotypes are 11.4%, 2.6%, and 3.1% respectively. Therefore we assume that the haplotypes of m1 -m2 are only T-A, C-A and C-G, but not T-G, as it were,there is no recombination happened between m1 site and m2 site. The frequencies of three haplotypes of T-A, C-A and C-G, calculated through corresponding genotypes, are 69.6%, 7.7% and 22.6% respectively. PMID- 15552038 TI - [The effect of HSV-tk/GCV on hepatic specific damage driven by murine ALB promoter/enhancer]. AB - The recombinant vector (pLLTK), containing murine serum albumin (ALB) gene promoter/enhancer directing the herpes simplex virus thymidine kinase (HSV-tk) gene expression was constructed to study its effect on hepatic cell-specific damage. Firstly,in order to compare the hepatic cell-specific transcriptional activity, three vectors were constructed in which green fluorescent protein (GFP) gene was used as a reporter marker. Vector pLE was driven by murine serum ALB gene promoter,whereas pLLE contained not only murine ALB promoter but also an enhancer located at upstream of the promoter was included. In the meanwhile, the vector pLEL had a murine ALB promoter and enhancer placed at the downstream from GFP. After transfected into Hep-G2 (a human hepatic cell line) and HC-11 (a murine breast epithelial cell line), GFP expression was examined by using fluorescence microscope as well as flow cytometer. Secondly, the vector pLLTK was used to observe the killing effect on Hep-G2 cells. The results demonstrated that ALB promoter/enhancer was able to direct hepatic cell-specific GFP expression. Furthermore, HSV-tk expression in Hep-G2 cells was ganciclovir (GCV)sensitive. After seven days of culture with GCV, pLLTK-transfected Hep-G2 cells showed an obvious cell death (53%) when detected by 3-4,5 dimethylthiozol-2-yl-2, 5 diphenyl tetrazolium bromide colorimetry (MTT) assay. Compared to the untreated group, there were no obvious changes in cellular growth inhibition rate in the Hep-G2 cells transfected with a blank control vector pcDNA3. 1 (only 2% of cells appeared cell death). All these results indicated that the above constructs were hepatocyte-specific. It therefore paves a way for further creating a liver specific damage animal model by transgenic approach using HSV-tk gene expression driven by the ALB promoter/enhancer. PMID- 15552039 TI - [Establishing mouse embryonic stem cell line carrying a fluorescent undifferentiated marker]. AB - To label mouse ES cells,a cell line derived from the inner cell mass of 3.5-day blastocysts,with enhanced green fluorescent protein (EGFP), the vector of pRex-1 EGFP was transferred into mouse ES cells by electroporation. The expressions of Rex-1 in undifferentiated and differentiated ES cells were detected by the microscopic observation of EGFP and by RT-PCR. The results showed that the EGFP gene was transferred into the mouse ES cell line, and the transfected cells in undifferentiated state showed high levels of EGFP expression. When the cells began to differentiate, the EGFP expressions were gradually reduced. A mouse ES cell line expressing EGFP under the control of Rex-1 gene promoter was generated. The cell line provides a powerful approach for the research of the process of mammalian development and for the screening of small molecules that can regulate this process. PMID- 15552040 TI - [Cloning of cDNA of SRG4, a mouse spermatogenesis related gene and expression in mouse different developing stages]. AB - Spermatogenesis required lots of genes taking part in this process. In prevision studies, Odf1, Odf2, Sak57, Spag4, Spag5 were identified in mouse and rat sperm tail developing. Two ESTs, BG101130 and BG100990, which coded amino acid sequences with high homology to rat Spag4, were found from mouse spermatocytes. By electrical hybridization, a 1 155 bp sequence was obtained and named as SRG4 (Spermatogenesis Related Gene 4) (GenBank accession number: AY307077), which included the full-length sequence of mouse testis hypothetical gene AK006225. The opening reading frame of SRG4 was 87 approximately 1 133 bp, and was proved by RT PCR in mouse testis. The information analysis showed SRG4 gene was located in 2H1 H2, and the putative protein was 348-amino acid with a coiled-coil region, suggesting SRG4 might function as a transmembrane protein. The homologies of amino acid sequences were 74% between TSARG4 and SRG4 gene and 45% between SRG4 and rat Spag4 gene, respectively. RT-PCR and Northern blot results showed SRG4 gene expressed specifically in mouse testis. Changes of SRG4 expression in mouse different developing stages were observed. The results showed SRG4 gene expressed slightly in two weeks postpartum, and expressed abundantly from three weeks, and reached the top at 4 -5 weeks. This indicted SRG4 gene may play an important role in mouse spermatogenesis. PMID- 15552041 TI - [Cloning and expression analysis in mature individuals of salmon gonadotropin releasing hormone (sGnRH) gene in common carp]. AB - Two types of complementary DNAs (cDNA) encoding the precursor of salmon gonadotropin-releasing hormone (sGnRH, [Trp7, Leu8] GnRH) are cloned and sequenced from common carp brain using rapid amplification of cDNA ends (RACE). The two cDNAs are referred to sGnRH cDNA1 and cDNA2, and the full-length fragment of cDNA1 and cDNA2 were 393 and 478 bp, respectively. Two sGnRH cDNAs contain an open reading frame of 285 bp, which encodes the sGnRH precursor including 94 amino acid residues. The sGnRH precursors consist of a signal peptide, sGnRH decapeptide and a GnRH-associated peptide (GAP) which is linked by the processing site (Gly-Lys-Arg). Two different sGnRH genes are characterized by intron trapping, and they share a similar structure composed of four exons and three introns. The nucleotide sequences identity of intron 1, intron 2 and intron 3 in sGnRH gene1 and gene2 are 71.1, 76. 1 and 88.0%, respectively. The basic structure and encoding architecture of sGnRH cDNAs and genes are similar with the reported GnRH. It is presumed that all the GnRH variants may evolve from a common ancestral molecular. Southern blot results confirm further the conclusion that there are two differential sGnRH genes in common carp genome. The results of reverse transcription-polymerase chain reaction (RT-PCR) assaying show that the two sGnRH genes co-express in the dissected brain regions, pituitary and ovary with the exception of expression of sGnRH genes in testis. According to the co existence of two sGnRH genes in distinct brain regions,pituitary and ovary, it is presumed that sGnRH plays an important role in the regulation of hypothalamic pituitary-gonadal (HPG) axis,and that sGnRH also operates as the neuromodulator, autocrine and/or paracrine regulator. PMID- 15552042 TI - [Three Bombyx mori genes, chi, gluE and fruA, encode proteins homologous to microorganism and primary analysis of horizontal gene transfer]. AB - According to the analysis of large scale EST sequencing of silkworm, Bombyx mori, we found that chi, glue and fruA of silkworm have very high homology at amino acid level and closely phylogenetic relative with that of microorganism, but lower similarity with genes of eelworm (Caenorhabditis elegans), fruitfly (Drosophila melanogaster), mosquito (Anopheles gambiae) and other relative insects, respectively. It indicates that each of them is likely to have common ancestor with that of microorganism. Namely, microbial genes were likely transferred to silkworm by horizontal gene transfer, instead of the vertical inheritance in evolutionary manner. PMID- 15552043 TI - Analysis of microsatellites derived from bee Ests. AB - To accelerate the molecular analysis of genetics,evolution and behavior, etc. in the honey bee (Apis), the frequency and density of simple sequence repeats (SSRs) have been analyzed from the bee EST (expressed sequence tag) database comprising 15 869 sequences amounting to 7.9 Mb. Results showed that the frequency of SSRs was 1/0.52 kb in bee ESTs, and hexanucleotide repeats (45.0%) motifs appeared to be the most abundant type in bee,the dinucleotide, mononucleotide, trinucleotide, tetranucleotide and pentanucleotide repeats are 17.9%, 14.1%, 11.6%, 9.2% and 2.2%, respectively. Meanwhile, the A-rich repeats are predominant in each type of SSRs, such as A, AT, AG, AC, AAT, AAG, AAC, AAAT, AAAG, AAAAG, AAAAT, AATAT, AAAAAG and AAAAAT repeats, whereas G-rich repeats are rare in the coding regions. The further analysis suggests that, apart from minor deviations, there is no significant difference in the distribution and density of microsatellites in the redundant and non-redundant set of bee ESTs. Furthermore, the availability of microsatellite markers can be expected to enhance the power and resolution of genome analysis in bee. PMID- 15552044 TI - [Isolation and expression of novel expressed sequence tags (ESTs) from ovarian follicles of Shaoxing ducks]. AB - Three expressed sequence tags ( ESTs), SXDF0201 (271 bp), SXDF0202 (200 bp) and SXDF0203 (173 bp), were isolated from ovarian follicles of Shaoxing ducks by using silver staining mRNA differential display. GenBank/BLAST analysis revealed that SXDF0201 was not homologous to any of the published sequences from all species, indicating that it was a novel EST and was then registered in GenBank (GenBank Accession No.: CB072629), while SXDF0202 and SXDF0203 were found to be highly homologous to seven known chicken ESTs and chicken mRNA for gizzard smooth muscle myosin heavy chain. 5'-RACE was employed to extend the SXDF0201 to 544 bp which was confirmed as novel in BLAST search. The temporal and spatial expression of SXDF0201 and SXDF0202 were also investigated with semi-quantitative RT-PCR. The result showed that: both SXDF0201 and SXDF0202 were found to be expressed in hypothalamus, pituitary, muscle, liver, and fat tissues of Shaoxing ducks; SXDF0201 was expressed significantly higher in ovaries of 30-day-old Shaoxing ducks compared with that of 60-day-old (P < 0.05) and 90-day-old (P = 0.015), but the expression of SXDF0202 showed no difference throughout the ovarian development; granulose layers expressed higher SXDF0201 than theca layers in almost all hierarchical follicles, the expression of SXDF0202 in granulose layers increased along with follicular maturation (P < 0.01) from Fw to F3 follicles, but decreased dramatically to the lowest in F1 follicles (P < 0.01). In theca layers, the highest expression of SXDF0202 was found in Fw follicles (P < 0.01). PMID- 15552045 TI - [Phylogenetic tree and synteny of DMRT genes family of vertebrates]. AB - Vertebrates contain a family of genes related to the Drosophila doublesex and C. elegans mab-3 genes, which encode transcription factors including a DNA-binding motif, DM domain. Evolution and function of different DMRT genes of vertebrates have not been understood yet,although some DM proteins are involved in sex determination, sexual differentiation and early embryonic development among different phyla. By genomic analysis of zebrafish and rat DMRT genes, all protein sequences of the vertebrate DMRTs were searched from gene databases and aligned. Phylogenetic tree of all these DMRT genes was reconstructed and evaluated by Bootstrap method. These DMRT genes were clustered into seven subfamilies. Results from analysis of gene structure and cluster organization of DMRT genes showed that synteny of DMRT genes of vertebrates were highly conserved among human, mouse, rat, fugu, medaka and zebrafish, with two syntenic groups, DMRT 1 approximately 3 and DMRT 5 approximately 6. PMID- 15552046 TI - [A new method for phylogenic analysis]. AB - With ESTs from porcine fatty tissue and cDNA sequences from human, bovine and mouse in non-reduncdant database and dbEST in GenBank,we sampled cDNA sequences of 70 function-known genes in four species on the base of randomly sampling method, analyzed the mutation pattern of 70 x 150 bp linking sequences between them, and established an integrated phylogenic analysis method. The results showed that 391 single bases mutations were found in 70 x 150 bp linking sequences alignment among four species. The number of mutation bases between them were greatly exceeded the 1/1000 predicted in the human genome analysis. C/T(T/C) and A/G (G/A) transitions were the major types of single base mutation. The genetic relationship between pig and bovine who are both Artiodactylous is the nearest, the next is human, and the farthest is mouse. The differentiation sequence taken place in four species from the same ancestor is that mouse is the earliest one, and the latter human, and pig and bovine are the latest. PMID- 15552047 TI - [Maximum likelihood analysis for mapping dynamic trait QTL in outbred population. I . Methodology]. AB - The quantitative traits whose phenotypic values change with time in life or other quantitative factors, were defined as dynamic traits. Based on the idea about random regression test-day model for estimating breeding values in animal evaluation, a mathematic model was constructed for mapping dynamic trait loci by using Legendre polynomials to model dynamic changes of each genetic effect. The Maximum likelihood analysis implemented via EM algorithm was used to estimate the parameters, including QTL position, fixed genetic regression effects for dynamic trait QTL mapping in outbred population. Compared with the existing method for dynamic trait mapping QTL, the new method presented here not only allowed to sample dynamic trait in disequillibrium way, but also can achieve to map dynamic traits QTL in any resource population by just one step. The further study on dynamic trait mapping QTL was theoretically discussed by incorporating genetic analysis of dynamic trait into general mapping QTL. PMID- 15552048 TI - [Identification of quantitative trait loci controlling plant height and days to heading from Yuanjiang common wild rice (Oryza rufipogon Griff.)]. AB - An advanced backcross strategy was used to identify quantitative trait loci (QTLs) associated with plant height and days to heading in the BC3F2 population derived from an accession of common wild rice (Oryza rufipogon Griff.) from Yuanjiang in Yunnan Province of China, as the donor, and an elite Indica cultivar "Teqing", as the recipient. Based on analyses of 116 SSR markers distributed throughout the rice genome and by using single-point analysis, four putative QTLs derived from O. rufipogon were detected for plant height on chromosome 1, and 6 QTLs for days to heading on chromosome 1, 3, 7, 8 and 11. All Alleles of QTLs for plant height from O. rufipogon could increase plant height of the backcross population. One QTL near RM104 on chromosome 1, explaining 27% and 28% of phenotypic variance and additive value reached 26.24 cm and 26.28 cm respectively in Beijing and Hefei, was detected. The location of this QTL is corresponding to sd-1. One QTL near the marker RM25 on chromosome 1, explaining 13% and 15% of phenotypic variance was identified. The additive value of this QTL reached 4.60 days and 3.65 days respectively in Beijing and Hefei, and the allele from O. rufipogon could delay the backcross population's days to heading. PMID- 15552049 TI - [Inheritance of bc1 gene in intersubspecific hybrids of rice (Oryza sativa L.)]. AB - Distorted segregation of the brittle culm-1 gene (bc1) on rice chromosome 3 was found with greatly increased or decreased frequency of bc1 bc1 genotype in inter subspecific hybrids, although the gene normally transmitted to its offspring following the Mendelian Law in intra-subspecific hybrids. In a combination of Kamairazu//Ketan Nangka/Kamairazu,an increased frequency of bc1 bc1 in F1, normal segregation in F2, and increased and decreased frequency in a few F3 and F4 lines were observed. In a cross of IR36/Kamairazu, decreased frequency in F2, both normal and decreased segregations in F3 and F4, and a few lines of increased ratio in F4 were found. In F2 of Ketan Nangka/IR36//Kamairazu, increased and decreased and normal segregations were all observed. There was no significant correlation between the frequency of bc1 bc1 and pollen fertility. It implied that distorted segregation of bc1 was caused by selective fertilization of male gametes, which were governed by gametophyte genes of ga2, ga3 and ga14 on chromosome 3. PMID- 15552050 TI - [Cloning of a MADS box protein gene (GhMADS1) from cotton (Gossypium hirsutum L.)]. AB - As a kind of transcription factors, MADS-box protein plays an important role in various cellular processes, especially in the development of floral organs. Based on the contig analysis of the cotton ESTs, the coding region of a cotton MADS-box protein (GhMADS1) was obtained by RT-PCR from floral buds of cotton (G. hirsutum). The cloned fragment of 713 bp (GhMADS1, GenBank accession no. AF538965) contains an open reading frame of 711 bp,coding a polypeptide of 236 amino acids. It was demonstrated that the deduced GhMADS1 protein was highly homologous to the AGL2 group of MADS-box proteins from Vitis vinifera, Nicotiana sylvestris, Petunia hybrida, Arabidopsis thaliana and Antirrhinum majus. Phylogenetic analysis also indicated that GhMADS1 belongs to the AGL2 group of MADS-box proteins. RT-PCR analysis showed that GhMADS1 gene expressed in petals, stamens, ovules and fibers, but not in roots, stems, leaves, bracts and sepals. The strongest expression of GhMADS1 gene was detected in petals. But in floral buds of a cotton homeotic mutant (CHV1), whose floral organs are all converted to bract leaf-like organs, the transcript of GhMADS1 gene was not detected. It was proposed that GhMADS1 gene would be crucial to the development of cotton floral organs. PMID- 15552051 TI - [Identifying the S genotypes of sweet cherry (Prunus avium L.) cultivars]. AB - This report identified S-RNase genes (S genes) of sweet cherry (Prunus avium L.), presented the sequences of S genes by using a pair of specific primers PruC2 and PruC4R based on the conserved regions C2 and RC4 of Rosaceous S-RNase genes and researched the S gene specific products from the genomic DNAs of different cultivars in which most of the S genotypese were unknown. The bands of PCR were cloned and their sequences were compared in GenBank. Four S genes were defined and the conclusion was made that all the same bands from PCR in the agarose gel had the same length and sequence of nucleic acid and were the same kind of S gene. The lengths of the amplified S genes are as follows: S1 is 677 bp, S3 762 bp, S4 945 bp, S6 456 bp. The S genotypes (S gene genotypes) of the tested self incompatible cultivars were identified as follows: 'Hongdeng', 'Hongyan' and 'Early ruby', as same as 'Van', were S1 S3; 'Jueze', 'Hongfeng' and 'Napoleon' were S3S4; 'Dazi' was S1 S6; 'Changbahong' was S1 S4; 'Elton' was S3S6. The self compatibile cultivars 'Waiyin No.7' and 'Stella' had the same S genotypes S3 S4'. PMID- 15552052 TI - [Constructing genetic linkage maps in Chinese Fir using F1 progeny]. AB - A new strategy was proposed for constructing linkage maps for 1:1 segregation molecular markers in Chinese fir. Linkage phase and recombination ratio for any two loci can be inferred and estimated. For the best order in a linkage group, the Hidden Markov Chain Model was used for multilocus linkage analysis. The mapping method proposed here was more effective than the 'pseudo-testcross' method commonly used in forest trees. In this paper,with the AFLP marker data obtained from the F1 progeny of 'Jurong 0' (female) x 'Rouye' (male), the genetic linkage maps of the Chinese fir clones, 'Jurong' and 'Rouye', were reconstructed based on the new mapping method. In the genetic linkage map of "Jurong 0", 101 markers were distributed in 11 linkage groups with an average distance of 22.6 cM and a total length of 2 282.6 cM. There were as many as 17 markers and as few as 5 markers in single linkage groups of the map. In the genetic linkage map of 'Rouye', 94 markers were distributed in the 11 linkage groups with an average distance of 27.3 cM and a total length of 2 565.8 cM. There were as many as 16 markers and as few as 4 markers in single linkage groups of the map. Compared with the 'pseudo- testcross' method, the two maps increased 26 markers and 28 markers, respectively, and totally 54 markers were added. The two maps contained 195 AFLP markers and each spanned more than 2 000 cM, almost covering the whole genome of Chinese fir. Therefore, the resolutions of linkage maps of Chinese fir 'Jurong 0' and 'Rouye' were improved with the new mapping method, and the maps provide the framework for further genetic analysis. PMID- 15552053 TI - [Cloning and functional analyses of promoters of two carbonic anhydrase genes from Dunaliella salina]. AB - The cloning vectors pMD-DCA1 and pMD-CA containing the promoters of duplicated carbonic anhydrase 1 (DCA1) and carbonic anhydrase (CA) genes, respectively, from Dunaliella salina, and expression vector pDM307 containing bar-NOS polyA fragment were digested with EcoR I. The bar-NOS polyA fragment was fused, respectively, to the fragments of the vectors pMD-DCA1 and pMD-CA to form transgenic D. salina expression vectors pMDDC-B and pMDC-B. The micro-shots were prepared by coating two constructs (pMDDC-B and pMDC-B) with gold particle. Each sample was bombarded once, twice, and thrice, respectively, with micro-projectile gun at a rupture pressure of 690 kPa in helium gas. The screening culture of the bombarded alga cells was performed in PKS liquid and solid medium containing 3 mg/L phosphinothricin (PPT) to develop the transformed cells of D. salina. Analyses of the transformed cells were carried out through PCR, Southern blotting, and Northern blotting. The results of screening culture showed that the expression of the external bar gene of vectors pMDDC-B and pMDC-B was stable and transient, respectively, in the transformed D. salina cells. In the meantime, the transformed efficiency of particle bombardment twice was higher than that of once or thrice particle bombardment at a rupture pressure of 690 kPa in helium gas. PCR and Southern blotting analyses indicated that the external bar gene was integrated into the genome of the cells. Northern analysis indicated that expression efficiency of the bar gene driven by DCA1 promoter was regulated by the gradient concentration of sodium chloride, and the positive blotting signal intensity of the bar mRNA was highest in the medium containing 2 mol/L of sodium chloride. The findings of the present study suggest that promoter of the DCA1 gene may be an inducible promoter following a hyperosmotic shock with high activity and safety in the research of transgenic D. salina. The tandem GT sequences of the promoter region of DCA1 and CA genes may be related to the molecular mechanisms of the extreme halo-toleration of the unicellular green alga, D. salina. PMID- 15552054 TI - [The research progress of zebrafish gene engineering]. AB - The research progress of zebrafish gene engineering is reviewed, in including the recent explosion status of transgenic zebrafish lines, transgeneic zebrafish in targeted screens, the development of transgenic technology in zebrafish as well as the current hotspots and future perspective of zebrafish gene engineering. By now the zebrafish gene engineering, nuclear transplantation and chromosome set manipulation technologies have been established in China. With firm foundation of zebrafish cytogenetics and embryology, we will obtain gene engineering zebrafish with gene targeting integration and promote the advancement and development of biotechnology in applied research field in near future. PMID- 15552055 TI - Ultrastructure of Gyrodinium spirale, the type species of Gyrodinium (Dinophyceae), including a phylogeny of G. dominans, G. rubrum and G. spirale deduced from partial LSU rDNA sequences. AB - A detailed ultrastructural analysis of the type species of Gyrodinium, G. spirale, was made based on cells collected from Skagerrak and southern Kattegat (Denmark). This material is considered very similar to the type material studied by Bergh from southern Kattegat. The analysis revealed many characters typical for dinoflagellates as well as a number of previously undescribed features. Here, emphasis was given to a three-dimensional configuration of the flagellar apparatus, the surface ridges, and the nuclear capsule. The latter had a rather complex ultrastructure consisting of two wall-like layers surrounded by membranes, with nuclear pores restricted to globular invaginations of these layers. To overcome difficulties with culturing of many auto- and heterotrophic dinoflagellates, we designed a specific reverse primer to amplify ca. 1800 base pairs of nuclear-encoded LSU rDNA. Using this approach, LSU rDNA sequences were determined from three heterotrophic species of Gyrodinium, including the type species. Using other alveolates (i.e. ciliates and Apicomplexa) as outgroup species, phylogenetic analyses based on Maximum Likelihood, Maximum Parsimony, and Neighbor-Joining supported Gyrodinium as a separate lineage. Unfortunately, the nearest sister group to Gyrodinium could not be established due to low bootstraps support for the deep branching pattern. PMID- 15552056 TI - The influence of mesozooplankton on phytoplankton nutrient limitation: a mesocosm study with northeast Atlantic plankton. AB - We used marine phytoplankton from mesocosms seeded with different zooplankton densities to study the impact of mesozooplankton on phytoplankton nutrient limitation. After 7 d of grazing (copepod mesocosms) or 9 d (appendicularian mesocosms) phytoplankton nutrient limitation was studied by enrichment bioassays. After removal of mesozooplankton, bioassay bottles received either no nutrients, phosphorus or nitrogen alone, or a combination of nitrogen and phosphorus and were incubated for 2 d. Phytoplankton reproductive rates in the bottles without nutrient addition were calculated after correction for grazing by ciliates and indicated increasing nitrogen limitation with increasing copepod abundance. No nutrient limitation was found in the appendicularian mesocosms. The increase of nutrient limitation with increasing copepod density seems to be mainly the result of a trophic cascade effect: Copepods released nanoplankton from ciliate grazing pressure, and thereby enhanced nitrogen exhaustion by nanophytoplankton and reduced nitrogen excretion by ciliates. Nitrogen sequestration in copepod biomass, the mechanism predicted by the ecological stoichiometry theory, seems to have been a weaker effect because there was only little copepod growth during the experiment. PMID- 15552057 TI - Paleoleishmania proterus n. gen., n. sp., (Trypanosomatidae: Kinetoplastida) from Cretaceous Burmese amber. AB - A trypanosomatid (Trypanosomatidae: Kinetoplastida) associated with a blood filled female sand fly in Cretaceous Burmese amber, is described in the new genus and species, Paleoleishmania proterus. The genus Paleoleishmania is established as a collective genus for digenetic fossil trypanosomes associated with sand flies. Amastigotes, promastigotes and paramastigotes are described. Paleoleishmania proterus is the first fossil kinetoplastid and provides a minimum age for the digenetic Trypanosomatidae. Its discovery indicates that vector-borne pathogens had been established by the Early Cretaceous. PMID- 15552058 TI - Sequence features of Oxytricha trifallax (class Spirotrichea) macronuclear telomeric and subtelomeric sequences. AB - We sequenced and analyzed the subtelomeric regions of 1356 macronuclear "nanochromosomes" of the spirotrichous ciliate Oxytricha trifallax. We show that the telomeres in this species have a length of 20 nt, with minor deviations; there is no correlation between telomere lengths at the two ends of the molecule. A search for open reading frames revealed that the 3' and 5' untranslated regions are short, with a median length of approximately 130 nt, and that surprisingly there are no detectable differences between sequences upstream and downstream of genes. Our results confirm a previously reported purine bias in the first approximately 80 nucleotides of the subtelomeric regions, but with this larger data set we curiously detected a 10 bp periodicity in the bias; we relate this finding to the possible regulatory and structural functions these regions must serve. Palindromic sequences in opposing subtelomeric regions, although present in most sequences, are not statistically significant. PMID- 15552059 TI - Characterization of chitin synthases from Entamoeba. AB - A major component of the Entamoeba cyst wall is chitin, a homopolymer of beta (1,4)-linked N-acetyl-D-glucosamine. Polymerization of chitin requires the presence of active chitin synthases (CHS), a group of enzymes belonging to the family of beta-glycosyl transferases. CHS have been described for fungi, insects, and nematodes; however, information is lacking about the structure and expression of this class of enzymes in protozoons such as Entamoeba. In this study, the primary structures of two putative E. histolytica CHS (EhCHS-1 and EhCHS-2) were determined by gene cloning and homologous proteins were identified in databases from E. dispar and the reptilian parasite E. invadens. The latter constitutes the widely used model organism for the study of Entamoeba cyst development. The two ameba enzymes revealed between 23% and 33% sequence similarity to CHS from other organisms with full conservation of all residues critically important for CHS activity. Interestingly, EhCHS-1 and EhCHS-2 differed substantially in their predicted molecular weights (73 kD vs. 114 kD) as well as in their isoelectric points (5.04 vs. 8.05), and homology was restricted to a central stretch of about 400 amino acid residues containing the catalytic domain. Outside the catalytic domain, EhCHS-1 was predicted to have seven transmembrane helices (TMH) of which the majority is located within the C-terminal part, resembling the situation found in yeast; whereas, EhCHS-2 is structurally related to nematode or insect chitin synthases, as it contained 17 predicted TMHs of which the majority is located within the N-terminal part of the molecule. Northern blot analysis revealed that genes corresponding to CHS-1 and CHS-2 are not expressed in Entamoeba trophozoites, but substantial amounts of CHS-1 and CHS-2 RNA were present 4 to 8 hours after induction of cyst formation by glucose deprivation of E. invadens. The time-courses of expression differed slightly between the two ameba CHS genes, as in contrast to CHS-1 RNA, expression of CHS-2 RNA was more transient and no plateau was observed between 8 and 16 hours of encystation. However, both CHS RNAs were no longer detectable after 48 hours when most of the cells had been transformed into mature cysts. PMID- 15552060 TI - Localization of centrins in the hypotrich ciliate Paraurostyla weissei. AB - Centrins are ubiquitous cytoskeletal proteins that are generally associated with the centrosome and form large cytoskeletal networks in protists. To obtain more data on the respective role of different centrin proteins, we studied their distribution and behavior in one ciliate species, Paraurostyla weissei, using specific antibodies. In this species, only two major proteins of 21 and 24 kDa corresponding to centrins, were identified by 1D and 2D electrophoresis. Immunofluorescence analysis showed that these two proteins displayed non overlapping localization in the interphase cell and during morphogenesis. Both centrin proteins localize on the fibrous network linking the oral basal bodies in the interphase cell and in the form of marginal dots, which correspond to the proximal ends of the striated rootlets; the 21 kDa centrin was also detected within the basal bodies, whereas the 24 kDa centrin allowed identifying new structures, the frontal dashes. During morphogenesis, the 21 kDa centrin locates at the basal bodies, while the 24 kDa centrin is detected along the striated rootlets and in close association with the basal bodies pairs. These data are discussed in terms of the potential roles of the two centrins in different cellular functions. PMID- 15552061 TI - Highly divergent SSU rRNA genes found in the marine ciliates Myrionecta rubra and Mesodinium pulex. AB - Myrionecta rubra and Mesodinium pulex are among the most commonly encountered planktonic ciliates in coastal marine and estuarine regions throughout the world. Despite their widespread distribution, both ciliates have received little attention by taxonomists. In order to better understand the phylogenetic position of these ciliates, we determined the SSU rRNA gene sequence from cultures of M. rubra and M. pulex. Partial sequence data were also generated from isolated cells of M. rubra from Chesapeake Bay. The M. rubra and M. pulex sequences were very divergent from all other ciliates, but shared a branch with 100% bootstrap support. Both species had numerous deletions and substitutions in their SSU rRNA gene, resulting in a long branch for the clade. This made the sequences prone to spurious phylogenetic affiliations when using simple phylogenetic methods. Maximum likelihood analysis placed M. rubra and M. pulex on the basal ciliate branch, following removal of ambiguously aligned regions. Fluorescent in situ hybridization probes were used with confocal laser scanning microscopy to confirm that these divergent sequences were both expressed in the cytoplasm and nucleolus of M. ruisra and M. pulex. We found that our sequence data matched several recently discovered unidentified eukaryotes in Genbank from diverse marine habitats, all of which had apparently been misattributed to highly divergent amoeboid organisms. PMID- 15552062 TI - International politics, Ectocarpus, Valonia, Halicystis, and Acetabularia. PMID- 15552063 TI - Have you an exit route? PMID- 15552064 TI - Funding and access to the GDS. PMID- 15552065 TI - Good occlusal practice. PMID- 15552066 TI - Good one, Watson. PMID- 15552067 TI - Is CPD effective? PMID- 15552068 TI - Uncemented crowns. PMID- 15552069 TI - Plasma cell gingivitis apparently related to the use of khat: report of a case. AB - Plasma cell gingivitis (PCG) is characterized by massive infiltration of plasma cells into the subepithelial tissue. It is a rare condition; the cause of which is still not fully understood. A case of PCG is reported in the mandibular gingiva probably caused by chewing khat. This report is the first, as far as we know, that relates PCG to the use of khat. The histological examination revealed infiltration of polyclonal plasma cells without signs of fungus, tuberculosis or malignancy. It is concluded that the changes were compatible with an allergic like reaction. The patient, a 30-year-old immigrant from Somalia, revealed in a subsequent consultation that he regularly used khat. The leaves are placed in the buccal sulcus. The PCG disappeared within two weeks of the use of khat being discontinued. Dental surgeons (periodontists) in Europe and the New World will, due to increasing immigration from Africa and the Arabian Peninsula, meet more patients who regularly use khat. This means that PCG and other khat related intraoral changes will become more common in the future. PMID- 15552070 TI - Crowns and extra-coronal restorations: endodontic considerations: the pulp, the root-treated tooth and the crown. AB - Endodontic considerations is the fourth in the series on crowns and other extra coronal restorations. This article focuses strongly on contemporary biological principles, and is not intended to provide a comprehensive review of commercially available materials and techniques. Principles are illustrated in a variety of clinical case scenarios. PMID- 15552071 TI - A randomised crossover trial of post-operative cognitive and psychomotor recovery from benzodiazepine sedation: effects of reversal with flumazenil over a prolonged recovery period. AB - OBJECTIVE: To study the post-operative cognitive and psychomotor recovery from midazolam conscious sedation, after reversal with the benzodiazepine antagonist flumazenil over a prolonged recovery period. DESIGN: A prospective, double-blind, randomised, crossover trial. SETTING: Out-patient Sedation Department, Newcastle Dental Hospital and School METHOD: Eighteen patients, ASA I or II, received midazolam on two separate occasions to undergo equivalent dental treatment. Following treatment patients were reversed with intravenous flumazenil or saline (placebo) at alternate appointments. Assessment of mood and cognitive function was undertaken using a highly sensitive and specific computerised battery of cognitive tests administered by telephone. Cognitive and psychomotor tests were administered prior to sedation and every hour for 6 hours post reversal. RESULTS: Results indicated no significant effect of flumazenil on simple reaction time and choice reaction time but did show a trend of reversing the effects of midazolam on numeric working memory and word recognition. CONCLUSION: The cognitive and psychomotor effects of the sedation were not fully reversed by flumazenil. Cognitive impairments were still present up to 6 hours post-reversal, despite patients appearing clinically more alert. This has important implications for treatment protocols and discharge instructions. PMID- 15552072 TI - Meta-analysis of measures of performance reported in oral cancer and precancer screening studies. AB - OBJECTIVE: To elicit a range of values for sensitivity, specificity and other measures of performance in screening for oral cancer and precancer. METHOD: A literature search which included three databases was conducted. Strict inclusion criteria were applied. Values for sensitivity (Sn) and specificity (Sp), from seven investigations, were expressed as a receiver operator characteristic (ROC) curve. Meta-analysis of the combined results was used to produce a summary operator characteristic (SROC) curve. RESULTS: The pooled weighted value of Sn from the seven studies was 0.796. From the SROC, the corresponding value of Sp at this level of Sn was 0.977 (95% CI 0.941, 0.991). When Sp was held at 0.977, the corresponding value of Sn from the SROC was 0.796 (95% CI 0.594, 0.912). CONCLUSIONS: The reports selected for eventual inclusion revealed a high level of heterogeneity with respect to the location of investigations, prevalence of lesions, the personnel used and other factors. The meta-analysis indicated that overall the studies had high discriminatory ability. The estimates of Sn and Sp, and values obtained for other measures of screening performance, were considered suitable for input to a simulation model in assessing the likely cost-utility of a variety of screening scenarios in further planned research. PMID- 15552073 TI - Orthodontic teaching practice and undergraduate knowledge in British dental schools. AB - AIM: The aim was to survey current orthodontic teaching practice in the undergraduate syllabus at British dental schools and to test the abilities of undergraduate students according to the requirements of the GDC regulations. MATERIALS AND METHODS: Information collected by means of a questionnaire sent to each dental school in 1998 was compared with similar data from 1994. The orthodontic knowledge and treatment planning ability of students was assessed by a multiple-choice examination paper completed by a random 10% sample of students from each dental school. RESULTS: In 1998 on average 195 curriculum hours were devoted to orthodontics and each student treated five patients. The teaching of fixed appliances had increased considerably between 1994 and 1998. The average MCQ score was 58% (range 39-72%). Students scored well on questions that tested basic knowledge but much less well when they were required to apply that knowledge. Only three schools felt that it was realistic to expect undergraduates to formulate orthodontic treatment plans, as they are required to do by the GDC. CONCLUSION: Results support the view that undergraduate orthodontic training should concentrate on diagnosis and recognition of problems rather than on providing limited exposure to treatment techniques. PMID- 15552074 TI - Where is the ethics in aesthetic dentistry? PMID- 15552075 TI - Focus Awards 2001. PMID- 15552076 TI - Biomagnetic localization of electrical current sources in the human heart with realistic volume conductors using the single-current-dipole model. AB - The boundary element method was applied in order to investigate the localization accuracy for focal sources measured from MCG data. Various homogeneous volume conductor models were composed: the individually shaped torso, a scaled standard torso, an unscaled standard torso, a scaled cuboid and a scaled ellipsoid. We implemented these models in single-dipole inverse solution techniques. High resolution multichannel data were analysed from two patients showing ventricular extrasystoles and two patients suffering from Wolff-Parkinson-White syndrome. Moreover, we report the localization of shallow- and deep-lying catheters (depth 9 cm and depth 17.5 cm below the measurement grid). Using an individually shaped homogeneous torso yields a localization error of less than 3 cm even for the deepest sources (mean error 2.4 cm). Probability-based dipole localization shows that the remaining error could only partly be explained by data noise statistics. Therefore it seems to be due to either inner inhomogeneities or the inadequacy of the single current dipole or a combination of the two. Thus clinically useful localization accuracy in the millimetre range requires more sophisticated volume conductor and source models. The evaluation of measurement data and simulation study shows that a scaled cuboid model can provide nearly the same localization accuracy as the individually shaped torso model. Single dipole reconstruction with this model is computationally faster than that with the individually shaped model of the human body and is fast enough for use in clinical applications. PMID- 15552077 TI - Photon beam convolution using polyenergetic energy deposition kernels. AB - In photon beam convolution calculations where polyenergetic energy deposition kernels (EDKS) are used, the primary photon energy spectrum should be correctly accounted for in Monte Carlo generation of EDKS. This requires the probability of interaction, determined by the linear attenuation coefficient, mu, to be taken into account when primary photon interactions are forced to occur at the EDK origin. The use of primary and scattered EDKS generated with a fixed photon spectrum can give rise to an error in the dose calculation due to neglecting the effects of beam hardening with depth. The proportion of primary photon energy that is transferred to secondary electrons increases with depth of interaction, due to the increase in the ratio mu ab/mu as the beam hardens. Convolution depth dose curves calculated using polyenergetic EDKS generated for the primary photon spectra which exist at depths of 0, 20 and 40 cm in water, show a fall-off which is too steep when compared with EGS4 Monte Carlo results. A beam hardening correction factor applied to primary and scattered 0 cm EDKS, based on the ratio of kerma to terma at each depth, gives primary, scattered and total dose in good agreement with Monte Carlo results. PMID- 15552078 TI - Analysis of photon beam exit dose using photon point kernels. AB - The Monte Carlo method is used to analyse the dose fall-off at the exit surface of a megavoltage photon beam. The convolution/superposition method of dose calculation using Monte-Carlo-generated homogeneous photon kernels is shown to be in error for exit dose calculation. Instead, photon kernels that incorporate modelling of the exit surface were generated, also using Monte Carlo, to analyse the problem, and the calculated dose fall-off using these kernels agrees well with measured data. In addition, the physics underlying the characteristics of the dose fall-off is analysed based on complete Monte Carlo modelling. Practical improvements to the convolution/superposition method are suggested. PMID- 15552079 TI - Dose-response curve slope improvement and result reproducibility of ferrous sulphate-doped gels analysed by NMR imaging. AB - Ferrous sulphate gel analysed by relaxation time measurements with NMR imaging is considered a useful dosimeter for 3D determinations of absorbed dose. A protocol for the gel preparation with agarose SeaPlaque that leads to a dosimeter with very high dose sensitivity is described. The dose-response curve slope is about 0.2 s(-1) Gy(-1) and the G factor turns out to be approximately 185 ions per 100 eV of absorbed energy. A method for making the measurements and analysing the results that brings about good result reproducibility is suggested. A thorough experimental study of the dependence of the dosimeter response on the elapsed time from preparation to irradiation and from irradiation to NMR measurement has revealed good reproducibility. The above characteristic of the gel system is very interesting, because it shows the possibility of utilizing the dosimeter for absolute dose determinations with satisfactory reliability. PMID- 15552080 TI - Quantitation in SPECT using an effective model of the scattering. AB - A new method for correcting simultaneously the attenuation, scatter and resolution effects in SPECT has been developed for the case of a homogeneous medium. It is based on an effective model of the scatter process. This model depends on only four parameters which are determined experimentally and remain independent of the geometry and of the dimensions of the scatter medium. The method uses the data from the peak events and does not need additional energy windows on the scattered events. An original filter is proposed to remove the noise due to the poor statistics of clinical data. Tests on phantoms varying in size and activity show that the model allows absolute activity determination with an accuracy of a few per cent. PMID- 15552081 TI - Noise and signal decoupling in maximum-likelihood reconstructions and Metz filters for PET brain images. AB - Images reconstructed with the maximum-likelihood-by-expectation-maximization (ML) algorithm have lower noise in some regions, particularly low count areas, compared with images reconstructed with filtered backprojection (FBP). The use of statistically correct noise model coupled with the positivity constraint in the ML algorithm provides this noise improvement, but whether this model confers a general advantage for ML over FBP with no noise model and any reconstruction filter, is unclear. We have studied the quantitative impact of the correct noise model in the ML algorithm applied to simulated and real PET fluoro-deoxyglucose (FDG) brain images, given a simplified but accurate reconstruction model with spatially invariant resolution. For FBP reconstruction, several Metz filters were chosen and images with different resolution were obtained depending on the order (1-400) of the Metz filters. Comparisons were made based on the mean Fourier spectra of the projection amplitudes, the noise-power spectra, and the mean region-of-interest signal and noise behaviour in the images. For images with resolution recovery beyond the intrinsic detector resolution, the noise increased significantly for FBP compared with ML. This indicates that in the process of signal recovery using ML, the noise is decoupled from the signal. Such noise decoupling is not possible for FBP. However, for image resolution equivalent to or less than the intrinsic detector resolution, FBP with Metz filters of various orders can achieve a performance similar to ML. The significance of the noise decoupling advantage in ML is dependent on the reconstructed image resolution required for specific imaging tasks. PMID- 15552082 TI - The evaluation of interactions between anaesthetic agent vapours and carrier gases by laser refractometry. AB - A measurement system and technique have been developed for the evaluation of interactions between anaesthetic agent vapours and carrier gases. The refractivities of mixtures of anaesthetic agent and carrier gas have been compared to values derived from the addition of the absolute refractivities of the individual components. These results show that there is no significant interaction between the anaesthetic agent and the carrier gas at the +/- 0.06% level of uncertainty. Results for the anaesthetic agents isoflurane, sevoflurane, enflurane, halothane, chloroform and desflurane and the carrier gases nitrogen, oxygen and dry air will be presented. PMID- 15552083 TI - Factors affecting penumbral shape and 3D dose distributions in stereotactic radiotherapy. AB - Linear accelerator based stereotactic radiotherapy or radiosurgery is usually performed with small collimated circular beams. The penumbral shape of these beams is caused by geometric penumbra and radiation penumbra; the former is affected by collimator position and focal spot size, the latter by energy and field size. This note reports measurements of penumbral width as collimator position, size and energy are varied. Radiation penumbra is a much greater effect than geometric penumbra at the distances used. The profile can also be changed by adding flattening filters to the external collimator. Adding a shaped filter to increase the primary radiation at the edges of the beam improves the shape of the profiles. What is of clinical importance is not the penumbral shape itself, but the effect it has on the three-dimensional dosage distributions. Cumulative dose volume histograms (DVHS) have been used to compare treatment plans produced by beams with different penumbral shapes. None of the aforementioned factors changes the DVH as much as a change in the field radius of a few mm. Ensuring that sufficient collimators are made, and reducing the interval between successive field sizes, is the single most important thing that can be done to avoid irradiating unnecessarily large volumes of tissue. PMID- 15552084 TI - Tibial lead determination by 99Tcm radiopharmaceutical x-ray fluorescence. AB - The feasibility of measuring tibial lead concentration by x-ray fluorescence with an internal 99Tcm labelled bone-seeking radiopharmaceutical was investigated using phantoms containing known values of lead concentration and 99Tcm activity. The minimum detectable concentration (MDC) at two standard deviations based on the counts in the Kalpha1 peak of 10.9 microg Pb ml(-1) was estimated to correspond to an MDC for an individual within a range of approximately 8-15 microg Pb (g bone mineral)(-1) if the counts from all four K x-ray peaks were included. Due to its much greater dose compared to an external source, the MDC of this internal source technique would have to be reduced before it could be used for measurements of occupationally or environmentally exposed individuals other than as an adjunct for a patient undergoing a bone scan. Methods of achieving such a reduction include increasing the acquisition time and the number of HPGE detectors, and optimizing the design of their collimators. PMID- 15552085 TI - Total-body irradiation on an isocentric linear accelerator: a radiation output compensation technique. AB - A treatment technique for total-body irradiation (TBI) is proposed that combines arc therapy with dynamic output control to achieve high-grade dose uniformity. The patient lies on a low couch and receives exposure in the prone and supine positions from a modulated arcing beam. The technique has been validated using a personal computer to control the linear accelerator and we demonstrate that only minor alterations to current dynamic therapy systems would be required. We have examined the practical application of this treatment with emphasis on methods of conformal therapy where an optimized dose distribution is prepared from a matrix of caliper measurements taken from the patient. This technique provides a means for regular TBI treatment on a computer-controlled linear accelerator that is easy to set up, requires short exposure times and is comfortable for the patient. PMID- 15552086 TI - An accurate energy-range relationship for high-energy electron beams in arbitrary materials. AB - A general analytical energy-range relationship has been derived to relate the practical range, Rp, to the most probable energy, Ep, of incident electron beams in the range 1 to 50 MeV and above, for absorbers of any atomic number. The expression is cubic in energy and requires as input parameters the total stopping power, So, the ratio of the scattering power and the total specific stopping power, T0/epsilon0, both taken at 10 MeV, and the radiation length for the material involved, X0. In addition to these parameters, five of the derived parameters are used to 'fine tune' the equation and minimize the mean square deviation from experimental and/or Monte Carlo data by means of non-linear regression. In the present study only Monte Carlo data determined with the new ITS.3 code have been employed. The standard deviations of the mean deviation from the Monte Carlo data at any energy are about 0.10, 0.12, 0.04, 0.11, 0.04, 0.03, 0.02 mm for Be, C, H2O, Al, Cu, Ag and U, respectively, and the relative standard deviation of the mean is about 0.5% for all materials. The fitting program gives some priority to water-equivalent materials, which explains the low standard deviation for water. A small error in the fall-off slope can give a different value for Rp. We describe a new method which reduces the uncertainty in the Rp determination, by fitting an odd function to the descending portion of the depth dose curve in order to accurately determine the tangent at the inflection point, and thereby the practical range. An approximate inverse relation is given expressing the most probable energy of an electron beam as a function of the practical range. The resultant relative standard error of the energy is less than 0.7%, and the maximum energy error deltaEp is less than 0.3 MeV. PMID- 15552087 TI - The theoretical and experimental evaluation of the heat balance in perfused tissue. AB - Accurate treatment planning is necessary for the successful application of hyperthermia in the clinic. The validity of four different bioheat models or combinations of models is evaluated: the conventional bioheat transfer equation, the limited effective conductivity model, a mixed heat sink-effective conductivity model and a discrete vessel model. The heat balance for the heated volume, and especially the ratio between conductive heat removal and heat escape through the veins, is different for each of these models. Model predictions were compared with results from experiments on isolated perfused bovine tongues. Tongues were suspended in a water-filled container and heated by conduction. The steady state temperature distribution and heat balance were determined at various blood flow rates. Increased blood flow was found to lower the mean tissue temperature and to enhance both conductive and venous heat removal. This result agrees only with the mixed heat sink-effective conductivity and the discrete vessel model predictions. At low flow rates a modified heat sink term should be used because the venous efflux temperature was significantly lower than the mean tissue temperature. PMID- 15552088 TI - Noise properties of the EM algorithm: I. Theory. AB - The expectation-maximization (EM) algorithm is an important tool for maximum likelihood (ML) estimation and image reconstruction, especially in medical imaging. It is a non-linear iterative algorithm that attempts to find the ML estimate of the object that produced a data set. The convergence of the algorithm and other deterministic properties are well established, but relatively little is known about how noise in the data influences noise in the final reconstructed image. In this paper we present a detailed treatment of these statistical properties. The specific application we have in mind is image reconstruction in emission tomography, but the results are valid for any application of the EM algorithm in which the data set can be described by Poisson statistics. We show that the probability density function for the grey level at a pixel in the image is well approximated by a log-normal law. An expression is derived for the variance of the grey level and for pixel-to-pixel covariance. The variance increases rapidly with iteration number at first, but eventually saturates as the ML estimate is approached. Moreover, the variance at any iteration number has a factor proportional to the square of the mean image (though other factors may also depend on the mean image), so a map of the standard deviation resembles the object itself. Thus low-intensity regions of the image tend to have low noise. By contrast, linear reconstruction methods, such as filtered back-projection in tomography, show a much more global noise pattern, with high-intensity regions of the object contributing to noise at rather distant low-intensity regions. The theoretical results of this paper depend on two approximations, but in the second paper in this series we demonstrate through Monte Carlo simulation that the approximations are justified over a wide range of conditions in emission tomography. The theory can, therefore, be used as a basis for calculation of objective figures of merit for image quality. PMID- 15552089 TI - Noise properties of the EM algorithm: II. Monte Carlo simulations. AB - In an earlier paper we derived a theoretical formulation for estimating the statistical properties of images reconstructed using the iterative ML-EM algorithm. To gain insight into this complex problem, two levels of approximation were considered in the theory. These techniques revealed the dependence of the variance and covariance of the reconstructed image noise on the source distribution, imaging system transfer function, and iteration number. In this paper a Monte Carlo approach was taken to study the noise properties of the ML-EM algorithm and to test the predictions of the theory. The study also served to evaluate the approximations used in the theory. Simulated data from phantoms were used in the Monte Carlo experiments. The ML-EM statistical properties were calculated from sample averages of a large number of images with different noise realizations. The agreement between the more exact form of the theoretical formulation and the Monte Carlo formulation was better than 10% in most cases examined, and for many situations the agreement was within the expected error of the Monte Carlo experiments. Results from the studies provide valuable information about the noise characteristics of ML-EM reconstructed images. Furthermore, the studies demonstrate the power of the theoretical and Monte Carlo approaches for investigating noise properties of statistical reconstruction algorithms. PMID- 15552090 TI - An estimation of noise levels in HMPAO RCBF SPECT images using simulation and phantom data; comparison with results obtained from repeated normal controls. AB - The purpose of this work was to determine noise levels in HMPAO RCBF SPECT images. Eight simulated images of a uniform sphere of activity were made at each of three different count levels. Three images of the Amersham brain phantom were obtained at each of three count levels, roughly corresponding to the simulation levels. Image reconstruction involved a modified Shepp-Logan filter with and without attenuation correction. The scaling constant in the Budinger equation was shown to vary little over the count range used with a mean value of 23 for uncorrected phantom data and 27 for corrected phantom data, corresponding to RMS noise levels of 7%-15%. The variance due to noise was calculated as a percentage of the variance obtained for 53 normal control studies following image registration and normalization. Values of 54% for uncorrected images and 67% for corrected images were obtained. For 10 normal controls a repeated study was performed. The ratio of within-subject to (single sample) between-subject variance was determined as 73% for uncorrected images and 78% for corrected images. PMID- 15552091 TI - A portable diffuse reflectance spectrophotometer for the rapid and automatic measurement of meat pigments. AB - A portable reflectance spectrophotometry system, which can measure a wide range of absorbance and has an adhesive pad type fibre optic probe, was developed to improve the availability of non-invasive reflectance spectrophotometry. A Czerny Turner type spectrometer, which can be connected to an optical fibre probe, was developed, and a linear charge coupled device (CCD) was used as its light detector to increase data acquisition speed. The scanning time of the linear CCD is automatically controlled in accordance with the reflectance of meat to increase the dynamic range of absorption. We tried to apply this instrument to samples of meat pigments in order to demonstrate whether reflectance spectrophotometry is suitable for evaluating the freshness of meat. We examined the myoglobin solution in the form of the met-derivative in order to detect the concentration of myoglobin. The absorbance spectrum of myoglobin in the range of 0-4 mg dl(-1) was successfully distinguished with this instrument. PMID- 15552092 TI - Teatment planning figures of merit in thermal and epithermal boron neutron capture therapy of brain tumours. AB - The boron neutron capture therapy (BNCT) figures of merit of advantage depth, therapeutic depth, modified advantage depth and maximum therapeutic depth have been studied as functions of 10B tumour to blood ratios and absolute levels. These relationships were examined using the Monte Carlo neutron photon transport code, MCNP, with an ideal 18.4 cm diameter neutron beam incident laterally upon all ellipsoidal neutron photon brain-equivalent model. Mono-energetic beams of 0.025 eV (thermal) and 35 eV (epithermal) were simulated. Increasing the tumour to blood 10B ratio predictably increases all figures of merit. concentration was also shown to have a strong bearing on the figures of merit when low levels were present in the system. This is the result of a non-10B dependent background dose. At higher levels however, the concentration of 10B has a diminishing influence. For boron sulphydryl (BSH), little advantage is gained by extending the blood 10B level beyond 30 ppm, whilst for D,L,-p-boronophenylalanine (BPA) this limit is 10 ppm. To achieve a therapeutic depth of 6 cm (brain mid-line from brain surface) using the thermal beam, a tumour to blood ratio of 25 with 10 ppm 10B in the blood is required for BPA. Similarly, a tumour to blood ratio of 8.5 with 30 ppm blood 10B is required for the maximum therapeutic depth of BSH to reach the brain mid-line. These requirements are five times above current values for these compounds in humans. Applying the epithermal beam under identical conditions, the therapeutic depth reaches the brain mid-line with a tumour to blood 10B ratio of only 5.7 for BPA. For BSH, the maximum therapeutic depth reaches the brain mid line with a tumour to blood ratio of only 1.9 with 30 ppm in the blood. Human data for these compounds are very close to these requirements. PMID- 15552093 TI - An algorithm for the fast registration of image sequences obtained with a scanning laser ophthalmoscope. AB - There is a need to quantify images of retinal pathology so that the natural progression and response to various forms of treatment such as laser therapy can be reliably monitored. The images obtained with a scanning laser ophthalmoscope are better than those from a traditional fundus camera, and the method of data collection leads naturally to digitization and quantitative techniques. However, during any angiographic sequence, there will inevitably be eye movement and it is essential that this be corrected for prior to the application of quantitative analysis. This article presents an algorithm (the smart localized binary correlation algorithm) for the automatic registration of fluorescein or indocyanine green angiograms which is based on localized correlation of binary templates in binary thresholded gradient images. The use of this algorithm decreases the calculation time up to 180 times when compared with the fastest applicable algorithm found in the literature. The algorithm only detects translational movement of the images but proves to be robust against rotational movement for the small angles (< 4 degrees) encountered in ophthalmic imaging. Registration accuracy is estimated to be +/- 2 pixels. PMID- 15552094 TI - Investigation of the imaging properties of an x-ray film scanner. AB - In this note, the imaging properties of a CCD x-ray film scanner were investigated. The relationship between optical density and the logarithm of output pixel value provides information on the linearity of the system. The resolution properties of the scanner can be characterized by the presampling modulation transfer function which was calculated from a set of line spread functions with various alignments relative to the sampling grid. Our results show that the scanner is linear up to 2.0 optical density units, which leads to restrictions in digitizing x-ray films with higher optical density. The measured MTF shows the good spatial resolution of the CCD scanner which is demonstrated on an example. PMID- 15552095 TI - Comments on 'A microdosimetric study of the dose enhancement in a fast neutron beam due to boron capture'. PMID- 15552096 TI - Solution of the linear bio-heat transfer equation. PMID- 15552097 TI - Aeq and other factors in a 60Co beam for a spherical or cylindrical minimal phantom and for an ionization chamber with a known A(wall) correction factor. AB - Three types of tissue-air ratio (TAR) are summarized. These TARS differ in their definition of the in-air absorbed dose. The first defines it as the absorbed dose at the centre of a spherical or cylindrical minimal water phantom in free space. The second defines it as the maximum primary absorbed dose in a semi-infinite water phantom. The third defines it as the absorbed dose in an imaginary infinitesimal mass of water within the cavity of a chamber in free space, where the absorbed dose is averaged within the cavity. It is concluded that the 60Co TAR data compiled by Godden and the 60Co TAR data of Johns and Cunningham should be reviewed. Aeq and other factors are evaluated for spherical and cylindrical minimal water phantoms in a 60Co gamma-ray beam. A method of obtaining Aeq and other factors for an ionization chamber with a known Awall correction factor is also reported. The work indicates some discrepancies with previously published material. PMID- 15552098 TI - Characteristics of 60Co gamma-ray SPR (scatter-primary ratio), SF (scatter factor), beta (dose-kerma ratio), and dmax (depth of maximum dose). AB - For 60Co gamma-rays, using zero-area tissue-maximum ratio (TMR) and revised scatter-maximum ratio (SMR) data, we investigate how the scatter-primary ratio (SPR), the scatter factor (SF), and the dose-kerma ratio (beta) change with field size and depth. We also investigate how the depth of maximum absorbed dose (dmax) changes with field size in three ways: the first uses zero-area primary plus scatter absorbed dose data, the second uses integrated primary absorbed dose data, and the third uses integrated primary plus scatter absorbed dose data. The investigated characteristics are also compared with reported ones. PMID- 15552099 TI - Midplane dose determination using in vivo dose measurements in combination with portal imaging. AB - The possibility of using portal films in combination with semiconductor in vivo measurements for midplane dose distribution is investigated. A general algorithm, using measured entrance and exit doses and available beam data of the Linac, is proposed to derive the midplane dose for symmetrical inhomogeneities. Experimental verification of the algorithm with phantom measurements is performed for different kinds of inhomogeneities (Al, air and cork) and phantom thicknesses from 13 cm to 30 cm. When using only the entrance dose and the exit dose, provided by the diodes on the beam axis, the algorithm predicts for the different inhomogeneities midplane doses in all cases within 1% of the midplane doses measured with an ionization chamber. When using the portal film in combination with entrance and exit dose measurements to estimate the midplane dose in an off axis position, the calculated midplane doses are within 3% of the midplane doses measured with an ionization chamber. The midplane doses calculated with the algorithm are compared to the midplane doses obtained with simplified calculation methods, i.e. the arithmetical mean and the geometrical mean of the measured entrance and exit doses. The geometrical mean especially seems to give acceptable results (within 5%) and can as such be used as an easy rule of thumb to estimate roughly the midplane dose. Finally, critical considerations on the validity and the precision of the proposed algorithm are given. The present results confirm the possibility of using the portal film for midplane dose distribution determination at the patient level. PMID- 15552100 TI - An algorithm to include the bremsstrahlung contamination in the determination of the absorbed dose in electron beams. AB - None of the existing protocols or codes of practice for high-energy electron dosimetry take any account of the accelerator-generated bremsstrahlung always present in electron beams. This results in a systematic error in the derivation of the absorbed dose. The purpose of this study is to draw attention to this omission which affects the absorbed dose calibration. A method based on available experimental data is presented for dealing with this deficiency in electron dosimetry. A re-defined algorithm for absorbed dose derivation accounting for this bremsstrahlung component is proposed. The question of omission of the bremsstrahlung contamination is important in comparing ionization methods with other dosimetric methods such as calorimetry or the use of ferrous sulphate. PMID- 15552101 TI - Iterative concurrent reconstruction algorithms for emission computed tomography. AB - Direct reconstruction techniques, such as those based on filtered backprojection, are typically used for emission computed tomography (ECT), even though it has been argued that iterative reconstruction methods may produce better clinical images. The major disadvantage of iterative reconstruction algorithms, and a significant reason for their lack of clinical acceptance, is their computational burden. We outline a new class of 'concurrent' iterative reconstruction techniques for ECT in which the reconstruction process is reorganized such that a significant fraction of the computational processing occurs concurrently with the acquisition of ECT projection data. These new algorithms use the 10-30 min required for acquisition of a typical SPECT scan to iteratively process the available projection data, significantly reducing the requirements for post acquisition processing. These algorithms are tested on SPECT projection data from a Hoffman brain phantom acquired with a 2 x 10(5) counts in 64 views each having 64 projections. The SPECT images are reconstructed as 64 x 64 tomograms, starting with six angular views. Other angular views are added to the reconstruction process sequentially, in a manner that reflects their availability for a typical acquisition protocol. The results suggest that if T s of concurrent processing are used, the reconstruction processing time required after completion of the data acquisition can be reduced by at least 1/3T s. PMID- 15552102 TI - Influence of window size in small-window colour measurement, particularly of teeth. AB - Tooth colour is often measured with a small window for illumination and measurement. This causes edge loss of the light, resulting in systematic errors in colour coordinates. This paper gives a quantification of the edge losses, and explains their cause. We measured reflectance spectra for 27 Formalin fixated extracted incisors using a small-window reflectance spectrophotometer equipped with external diaphragms of 3, 4, and 5 mm diameter, and using a spectroradiometer. We calculated the colour coordinates L*a*b* from these spectra. Finally, 16 randomly chosen teeth were illuminated with a pencil beam (lambda = 543 nm, and lambda = 633 nm) while the emerging light was measured as a function of distance from the illuminated spot using a CCD detector. These data were used to calculate small-window edge losses, and thus to predict the small window reflectance factors relative to spectroradiometrically determined reflectance, at both 543 nm and 633 nm. In all instruments the same spot on the tooth was illuminated and measured, and the teeth were always wet. Colour coordinates for the small-window colour measurements deviate significantly from those determined using the spectroradiometer. These deviations can be explained from the wavelength-dependant edge loss that arises in small-window colour measurement. PMID- 15552103 TI - The effect of alterations in physical and chemical characteristics on TOBEC derived body composition estimates: validation with non-human models. AB - The measurement of total-body electrical conductivity (TOBEC) has become one of the standard methods for the estimation of body composition in infants. We investigated, using non-human models, the effect on the accuracy of TOBEC-derived body composition estimates of alterations in physical and chemical characteristics of the fat-free mass (FFM). The effect of electrolyte type, concentration and volume on TOBEC was determined using 2, 3 and 51 solutions of six different chlorides and sodium bicarbonate. Equimolar concentrations yielded TOBEC values in accordance with known ion conductivities: H+ >> Ca2+ > Mg2+ > K+ > Na+ > Li+ and Cl- > HCO3-. The behaviour of these solutions was described very accurately over a wide range of concentrations (1-200 mM) by a simple exponential law. Dissolved egg-white protein, glycine and L-glutamine elicited no TOBEC signal. In vitro, using polyethylene bottles filled with physiologic saline, in the interval of 2-45 degrees C a linear relation was observed between temperature and TOBEC. Below the freezing point no TOBEC signal was elicited. The effect of tissue autolysis and body temperature on TOBEC was examined by repeated measurements of TOBEC and temperature in seven fresh infant minipig cadavers. Five minipigs were allowed to cool. Shortly after death TOBEC decreased by 2.5% per degrees C. Two animals were kept at constant temperature. The TOBEC signal showed a gradual increase of 9% after 7 h due to autolysis. We conclude that in vivo TOBEC measurements are affected by ion concentration (e.g. non-isotonic hydration changes), geometry (e.g. deviations in body shape), temperature (e.g. fever, skin cooling) and tissue autolysis (measurements after death). Proteins, molecules with strong dipole moments and ions trapped in crystalline structures do not significantly affect the TOBEC reading. PMID- 15552104 TI - Frequency-domain optical absorption spectroscopy of finite tissue volumes using diffusion theory. AB - The goal of frequency-domain optical absorption spectroscopy is the non-invasive determination of the absorption coefficient of a specific tissue volume. Since this allows the concentration of endogenous and exogenous chromophores to be calculated, there is considerable potential for clinical application. The technique relies on the measurement of the phase and modulation of light, which is diffusely reflected or transmitted by the tissue when it is illuminated by an intensity-modulated source. A model of light propagation must then be used to deduce the absorption coefficient. For simplicity, it is usual to assume the tissue is either infinite in extent (for transmission measurements) or semi infinite (for reflectance measurements). The goal of this paper is to examine the errors introduced by these assumptions when measurements are actually performed on finite volumes. Diffusion-theory calculations and experimental measurements were performed for slabs, cylinders and spheres with optical properties characteristic of soft tissues in the near infrared. The error in absorption coefficient is presented as a function of object size as a guideline to when the simple models may be used. For transmission measurements, the error is almost independent of the true absorption coefficient, which allows absolute changes in absorption to be measured accurately. The implications of these errors in absorption coefficient for two clinical problems--quantitation of an exogenous photosensitizer and measurement of haemoglobin oxygenation--are presented and discussed. PMID- 15552105 TI - Effect of anisotropy corrections on the dynamic dose calculations in high dose rate (HDR) brachytherapy. AB - An analytic formalism utilizing a convolution technique is presented for the incorporation of anisotropy corrections into the dynamic (or transit) dose calculations in high dose rate (HDR) brachytherapy. A simple numerical implementation of the suggested formalism is also presented. Two calculational examples are provided in order to show some effects of anisotropy corrections on the dynamic dose calculations. Based on the results from these examples and published literature, the absence of anisotropy corrections in the dynamic dose calculations could result in a dosimetric error up to the order of 0.1 Gy during the full course of HDR brachytherapy treatments depending on fractionation and geometrical arrangement. Therefore, the incorporation of anisotropy corrections into dynamic dose calculations could eliminate this kind of error in HDR brachytherapy. PMID- 15552106 TI - Modelling induced currents in biological cells exposed to low-frequency magnetic fields. AB - Interactions of low-frequency magnetic fields with biological systems have been a subject of intense scientific inquiry and public concern. Most research has been done at powerline frequencies of 50 Hz or 60 Hz. One of the key questions related to interactions of low-frequency magnetic fields with biological systems is which parameters of the exposure field are responsible for observed effects. Knowledge of the induced electric field and current in various experimental in vitro systems is important for this purpose. The 3D impedance method is used in this research to model spatial patterns of induced electric fields and current in two preparations of cells. A cell monolayer with a random distribution of cells and a confluent monolayer of cells with gap junctions are considered; because of the limitations of the computational method, biological cells are represented by cubes rather than more realistic shapes (e.g. spheres). The random model indicates that for higher cell densities the pattern of the induced current flow has a limited dependence on the size and shape of the container in which the cells are placed, it depends mostly on the actual cell placement. Gap junctions, not surprisingly, are shown to increase the current density, but only if their resistance is sufficiently low. The highest current density occurs in the gaps. PMID- 15552107 TI - Laminated primary ceiling barriers for medical accelerator rooms. AB - In this work the performance of a number of laminated shields used as the primary ceiling barrier for high-energy accelerator rooms was investigated. The neutron and photon dose equivalent rates were measured outside each shield. Based on the measured photon level and the calculated x-ray leakage level the dose equivalent rate due to photons produced by neutron interactions with the shield materials was estimated. Shielding parameters for polyethylene were established for photons and neutrons. It was found that the barriers designed using the techniques given in an NCRP Report suffered from excessive radiation leakage. PMID- 15552108 TI - Dose distribution measurements in superficial x-ray beams using NMR dosimetry. AB - The conversion of Fe2+ to Fe3+ in a Fricke solution due to ionizing radiation can be detected using nuclear magnetic resonance (NMR) imaging. The aim of the present study was to develop a suitable system for the study of dose distributions in superficial radiation beams making use of the water equivalence of the dosimetric gel system. Agarose gels (1.5%) doped with 0.5 mM ammonium ferrous sulphate and 125 mM sulphuric acid were exposed to x-rays from a superficial radiotherapy treatment unit (HVT 1.4, 2.4 and 7.25 mm A1). In the experiments doses between 10 Gy and 20 Gy were given, and the gel surface was in direct contact with the lead glass cone applicators (diameters 1 cm and 2.5 cm) at an FSD of 10 cm. Images were obtained within 4 h after irradiation in the head coil of a 1.5 T clinical MR scanner. Using spin-echo sequences with seven different repetition times between 120 ms and 4 s, the spin-lattice relaxation time (T1) was calculated for specified regions of interest with about 1 mm spacing. The inverse of T1 was shown to be proportional to the given dose and 1/T1 maps were obtained for all three superficial radiation qualities. The depth dose curves determined with NMR dosimetry compare well with those obtained with a thin-window parallel plate ion chamber and thermoluminescence dosimetry in the same radiation beams. PMID- 15552109 TI - Numerical calculation of energy deposition by high-energy electron beams: III. Three-dimensional heterogeneous media. AB - The phase space time evolution model of Huizenga and Storchi and Morawska Kaczynska and Huizenga has been modified to accommodate calculations of energy deposition by arbitrary electron beams in three-dimensional heterogeneous media. This is a further development aimed at the use of the phase space evolution model in radiotherapy treatment planning. The model presented uses an improved method to control the evolution of the phase space state. This new method results in a faster algorithm, and requires less computer memory. An extra advantage of this method is that it allows the pre-calculation of information, further reducing calculation times. Typical results obtainable with this model are illustrated with the cases of (i) a 20 MeV pencil beam in a water phantom, (ii) a 20 MeV 5 x 5 cm2 beam in a water phantom containing two air cavities, and (iii) a 20 MeV 5 x 5 cm2 beam in a water phantom containing an aluminium region. The results of the dose distribution calculations are in good agreement with and require significantly less computation time than results obtained with Monte Carlo methods. PMID- 15552110 TI - A fast and sensitive TLD method for measurement of energy and homogeneity of electron beams using transmitted radiation through lead. AB - A simple and fast, but sensitive TLD method for the measurement of energy and homogeneity of therapeutically used electron beams has been developed and tested. This method is based on the fact that when small thicknesses of high-Z absorbers such as lead are interposed in the high-energy electron beams, the transmitted radiation increases with the energy of the electron beams. Consequently, the ratio of readouts of TLDS held on the two sides of a lead plate varied sharply (by factor of 70) with a change in energy of the electron beam from 5 MeV to 18 MeV, offering a very sensitive method for the measurement of the energy of electron beams. By using the ratio of TL readouts of two types of TLD ribbon with widely different sensitivities, LiF TLD-700 ribbons on the upstream side and highly sensitive CaF2:Dy TLD-200 ribbons on the downstream side, an electron energy discrimination of better than +/- 0.1 MeV could be achieved. The homogeneity of the electron beam energy and the absorbed dose was measured by using a jig in which the TLDS were held in the desired array on both sides of a 4 mm thick lead plate. The method takes minimal beam time and makes it possible to carry out measurements for the audit of the quality of electron beams as well as for intercomparison of beams by mail. PMID- 15552111 TI - Beam characteristics of the therapax DXT300 orthovoltage therapy unit. AB - The operating performance and beam characteristics of a new orthovoltage unit, the Therapax DXT300, have been evaluated. Percentage depth-dose and backscatter tables are presented for several applicator sizes, at 30 cm and 50 cm focal skin distances (FSDS) and for multiple x-ray beam qualities with the tube operating between 100 and 300 kVp accelerating potential. The unit has been found to provide beam characteristics similar to those reported for other orthovoltage therapy machines. The linearity and short- and long-term stability/reproducibility of the unit's internal dosimetry system have also been studied, and results indicate a very stable beam output of better than 1% standard deviation. The data presented in this work should provide the basis for comparison with other units and act as a reference for clinics commissioning the Therapax DXT300 in the future. PMID- 15552112 TI - A comparison of four ultraviolet sources to alter graft-versus-host responses. AB - Graft-versus-host disease (GVHD) is a major contributor to the morbidity and mortality associated with allogeneic bone marrow transplantation. Direct ultraviolet B (UVB) irradiation of bone marrow and spleen cell allografts in mice using broadband lamps is known to abolish alloreactive responses which would normally cause GVHD. Using a histoincompatible murine model, we have extended these observations by comparing the physical spectrum of four UV sources (the Philips TUV8W, TL12 and TL01, and the Spectronics XX15B) with in vitro assessment of bone marrow progenitor cell damage and suppression of lymphocyte proliferation and in vivo comparison of the effect on GVHD of the TL12 and XX15B and on the rate of engraftment with the TL12. At doses of uv found to abolish lymphocyte proliferation (2.5, 7, 12 and 1000 J m(-2) with the TUV8W, XX15B, TL12 and TL01 lamps) colony-forming unit granulocyte-macrophage (CFU-GM) proliferation was reduced to 81%, 71%, 79% and 62%, respectively. At an optimal dose found to suppress GVHD (100 J m(-2) integrated radiant energy from 200-320 nm for the TL12 and XX15B) CFU-GM proliferation showed a reduction of 98% with the XX15B and 86% with the TL12. At this radiant energy with the TL12, the rate of bone marrow engraftment was impaired with 72% marrow cellularity at 2 weeks, decreasing to 48% after 200 J m(-2). Our results with this model demonstrate that broadband UVB irradiation of bone marrow permits transplantation across a major histocompatibility barrier. Furthermore we have provided in vitro evidence that narrowband UVB or UVC might potentially be applied to this model. PMID- 15552113 TI - Quality assessment of DSA, ultrasound and CT digital images compressed with the JPEG protocol. AB - Digital diagnostic images from subtraction angiography, computer tomography and ultrasound grabbed at 640 x 560 pixel and 320 x 280 pixel, eight-bit resolution were compressed at seven different levels with the 'lossy' JPEG algorithm in order to decrease the computer magnetic memory space required for their storage. After reconstruction, the deformations caused by the process were assessed (i) qualitatively: a perception test was performed twice, by five radiologists, on a total of 14 images compressed at various levels and (ii) quantitatively: the average grey level value per pixel was calculated at each compression level from each system's residual images obtained by subtracting the compressed image from the original and taking the absolute value of the result. It was concluded that images compressed at a ratio of about 10:1 were indistinguishable from the originals and hence could be considered acceptable for clinical use. PMID- 15552114 TI - A fast non-invasive beam check for mammography x-ray units. AB - A prototype instrument is presented for the indirect evaluation of the main physical parameters characterizing a mammographic x-ray beam. The instrument consists of an x-ray probe with four filtered solid-state detectors connected to a signal-conditioning electronic circuitry and a high-speed digitizer interfaced to a portable personal computer. The system, governed by an ad hoc developed software, can measure kVp, kV waveform, ripple, HVL, exposure time and exposure in a single shot of the x-ray beam with good accuracy. The system should prove very useful to any quality-control programme for mammography. PMID- 15552115 TI - Magnetic resonance imaging of radiation dose distributions using a polymer-gel dosimeter. AB - A new formulation of a tissue-equivalent polymer-gel dosimeter for the measurement of three-dimensional dose distributions of ionizing radiation has been developed. It is composed of aqueous gelatin infused with acrylamide and N, N'-methylene-bisacrylamide monomers, and made hypoxic by nitrogen saturation. Irradiation of the gel, referred to as BANG, causes localized polymerization of the monomers, which, in turn, reduces the transverse NMR relaxation times of water protons. The dose dependence of the NMR transverse relaxation rate, R2, is reproducible (less than 2% variation) and is linear up to about 8 Gy, with a slope of 0.25 s(-1)Gy(-1) at 1.5 T. Magnetic resonance imaging may be used to obtain accurate three-dimensional dose distributions with high spatial resolution. Since the radiation-induced polymers do not diffuse through the gelatin matrix, the dose distributions recorded by BANG gels are stable for long periods of time, and may be used to measure low-activity radioactive sources. Since the light-scattering properties of the polymerized regions are different from those of the clear, non-irradiated regions, the dose distributions are visible, and their optical densities are dependent on dose. PMID- 15552116 TI - A technique for simulation of the point spread function of a gamma camera. AB - A technique for simulating the point spread function (PSF) of gamma-camera images which treats primary photons and detected scattered photons separately is described. The primary photon PSF was measured at different distances in air from the camera. The scatter PSF was also assessed with different thicknesses of water between the point and the camera, and fitted to a circularly symmetric bi exponential function. An analysis of the variation of detected scattered radiation based on the results is presented. Simulation of the PSF at a given distance from the camera and thickness of attenuating medium was then possible from the sum of the appropriate primary photon PSF corrected for attenuation and the appropriate scatter PSF. The accuracy of the technique has been studied with variation in depth, density and uniformity of thickness of the attenuating medium. PMID- 15552117 TI - Quantitative brain FDG/PET studies using dynamic aortic imaging. AB - Positron emission tomography (PET) measurements of cerebral glucose utilization using 18F-fluorodeoxyglucose (FDG) are a useful tool in the investigation of localized brain function in normal and disease states. A major impediment to the application of FDG/PET in clinical investigation has been the need for arterial blood sampling to quantify cerebral glucose metabolism (CMRGlc). Qualitative studies, though informative in a variety of clinical settings, are of limited value for research applications and do not utilize the inherent quantitative nature of PET. We present a novel PET technique employing a whole-body PET tomograph with abdominal aortic imaging from 0 to 30 min as an alternative to arterial blood sampling to obtain the input function for cerebral metabolic rate calculations. Two or three arterial samples taken during the 10-45 min period were used to scale and extend the blood curve and the brain was imaged from 35-55 min post-injection. We performed 12 studies in which both arterial blood sampling and aortic scans were obtained. We found the correlation of global metabolic rates (GMR) when comparing the two techniques to be extremely high (R2 = 0.99). This suggests that the use of dynamic aortic imaging is less invasive and a viable alternative to arterial blood sampling in quantitative FDG/PET imaging. PMID- 15552118 TI - Effects of slow flow on slice profile and NMR signal in fast imaging sequences. AB - A computer program has been developed to evaluate the selective-slice profiles obtained in the steady state for fast gradient-echo imaging. Both spoiled and refocused gradient-echo pulse sequences have been considered. By numerically solving the Bloch equations modified for the effects of flow, for a three dimensional volume of spins, for realistic RF excitations and linear gradient combinations, the program permits the combined effects of flow and imaging variables on the magnetization slice profile to be assessed quantitatively. We have found that the gradient pattern in gradient-echo pulse sequences is a significant factor for determining the steady-state slice profiles and the strength of the NMR signal from the flowing spins. PMID- 15552119 TI - An investigation of an electromagnetic method for the measurement of body composition. AB - This note presents an investigation of an electromagnetic method for the measurement of the volume of electrolytes in the human body. The shift in the frequency of a resonant transmission line, wound as a large helix, when a phantom containing 1% saline was inserted coaxially, was found to be proportional to the volume of saline for small volumes only and for larger volumes became proportional to the surface area. This was observed for twophantom geometries. The volume of saline used ranged from 10 ml to 60 l. These observations suggest that the resonant electromagnetic system only responds to water at the surface. PMID- 15552120 TI - A phantom for the testing and calibration of near infra-red spectrometers. AB - This note describes a simple phantom which can be used to check the long term stability and accuracy of relative attenuation measurements made by near infra red (NIR) spectrometers. The phantom has been calibrated, and the results of the calibration are shown to agree with theoretical predictions of the relative attenuation. PMID- 15552121 TI - Photon backscattering tissue characterization by energy dispersive spectroscopy evaluations. AB - Techniques for in vivo tissue characterization based on scattered photons have usually been confined to evaluating coherent and Compton peaks. However, information can also be obtained from the energy analysis of the Compton scattered distribution. This paper looks at the extension of a technique validated by the authors for characterizing tissues composed of low-atomic-number elements. To this end, an EDXRS (energy dispersive x-ray spectrometry) computer simulation procedure was performed and applied to test the validity of a figure of merit able to characterize binary compounds. This figure of merit is based on the photon fluence values in a restricted energy interval of the measured distribution of incoherently scattered photons. After careful experimental tests with 59.54 keV incident photons at scattering angles down to 60degrees, the simulation procedure was applied to quasi-monochromatic and polychromatic high radiance sources. The results show that the characterization by the figure of merit, which operates satisfactorily with monochromatic sources, is unsatisfactory in the latter cases, which seem to favour a different parameter for compound characterization. PMID- 15552122 TI - The design and performance characteristics of a multileaf collimator. AB - A multileaf collimator, which has been in routine clinical use for both conventional and conformal radiotherapy for over four years, is described in detail. The collimator replaces the conventional treatment head of a Philips SL series linear accelerator and comprises 80 tungsten leaves and two orthogonal pairs of back-up collimators. Each leaf projects a width of 1 cm in the isocentric plane, allowing shaped photon treatment beams of up to 40 cm square. The performance of the prototype and first production model have been thoroughly tested against the design specifications and the requirements of IEC standards. Radiation attenuation by the collimator components has been measured and substantially exceeds those requirements. The irregular portion of a field (shielded by the leaves only) receives, on average, a dose of less than 2% of the tumour dose. The effect on the penumbra of using leaves which translate linearly and have curved faces has been assessed and found not to degrade the sharpness of the beam fall-off significantly. The reproducibility of the video system used in positioning the leaves has been measured and gave a root mean square deviation of less than 0.3 mm in repeat setting of a 10 cm square field, and an accuracy always within 1 mm. The rationale for clinical use of the device is discussed and its effect on treatment quality control and reliability, is considered. PMID- 15552123 TI - A new scintillator detector system for the quality assurance of 60Co and high energy therapy machines. AB - A new single-channel detector system has been developed to perform routine quality assurance of 60Co and high-energy therapy machines. This detector is composed of an orange plastic scintillator, optically coupled to a radiation resistant polycarbonate light pipe and a shielded silicon photodiode imbedded in a hollow solid water phantom block. No temperature and pressure corrections are required. Stability results were consistent with standard deviations fluctuating from 0.03% up to 0.09% for 60Co and from 0.05% up to 0.18% for other high energies. This device provides a quick, easy and reliable beam output check remotely, using an automatic reset based on a radiation triggering system device, storing multiple sequential readings. The reproducibility of this detector was checked on a daily and weekly basis at different energies (60Co, 6 MV and 18 MV x rays and 6, 9, 12, 16 and 20 MeV electron beams). These results were found to be consistent with those obtained using an ion chamber. Other characteristics of this detector, including the consequences of the radiation-induced light in the light pipe (stem effect) and the radiation damage on this system are briefly discussed. PMID- 15552124 TI - Quantitative imaging of holmium-166 with an Anger camera. AB - The objective of this study was to develop a quantitative Anger camera imaging approach for 166Ho in the skeletal system of patients. A dual energy window method was designed to subtract the interference from septal penetration and bremsstrahlung events in Anger camera images acquired with the 80 keV x-rays emitted by 166Ho. The validity of this scatter subtraction method for 166Ho images was demonstrated as improvements of the line spread function and modulation transfer function. Camera sensitivity was found to be nearly independent of source-to-collimator distance only for images acquired with a high energy collimator. Studies in an Alderson abdominal water phantom demonstrated scatter subtraction can provide quantitative Anger camera images of 166Ho with a scatter multiplier of k = 1.0 and a correction for attenuation. Attenuation correction factors derived from a transmission image were measured for the phantom and verified with water-equivalent blocks of known thickness. Whole-body scan images of 166Ho localized in the skeletal system of patients were significantly improved with this simple scatter subtraction method, and when used to estimate the activity distribution within separate bone regions of the skeleton. PMID- 15552125 TI - An analytical reconstruction algorithm for multifocal converging-beam SPECT. AB - Multifocal converging-beam collimation has been suggested for cardiac SPECT imaging to increase sensitivity over the heart without truncation of the activity distribution in the chest. In this study, an analytical reconstruction algorithm is derived for multifocal fan-beam and multifocal cone-beam tomography. In the algorithm, the projection data are differently weighted and filtered, depending on the distance from the detector. For a given image point, the set of filtered data corresponding to the distance between this point and detector is backprojected to determine the pixel value of the point. Thus, the backprojection is done only once at each projection view. To evaluate the algorithm, simulation studies are performed using a 3D Defrise slab phantom without considering photon attenuation and scatter, detector response, and statistical noise. Reconstructed images demonstrate that reasonable quality can be achieved with a modest focal length variation rate and with a small radius of rotation. A collimator with a focal length increasing quickly near its centre provides better quality in the image region distant from the central plane of the cone geometry, but produces more severe artifacts at the centre of the reconstructed image, compared to a collimator with an initially slowly varying focal length. PMID- 15552126 TI - An ice calorimeter for photon dosimetry. AB - A prototype absorbed-dose ice calorimeter has been constructed of ice made with triple-distilled water. The calorimeter was formed by sandwiching a thermistor between two blocks of ice. Initial measurements in a 60Co gamma-ray beam show that ice calorimetry is feasible, giving a signal to noise ratio of about 300 for a dose of 5 Gy. PMID- 15552127 TI - Polarity effect of plane-parallel ionization chambers in electron radiation. AB - Although plane-parallel ionization chambers have been in use for some time, there is still much to be learned about their performance characteristics. This work is concerned with the polarity effect in electron beams about which there is little published data. The investigations involved several popular ionization chambers; the PTW Markus chamber, the NACP chamber and its Calcam version, the Vinten-631 chamber, and the NE 2571 (Farmer-type) thimble chamber. The chambers were irradiated in electron beams of nominal energies between 4 MeV and 18 MeV. It was found in this study that the NACP, Markus and Vinten chambers require a correction of the order of 0.2% in the energy range between 4.5 MeV and 18 MeV. The overall behaviour of the Calcam chamber was similar with the exception of energies below 4 MeV. The depth dependence of the polarity effect seemed closely related to the mean beam energy at the depth of measurement. There is some evidence that the effect is also dependent on the angular spread of the electron beam and its spectrum. The authors considered how best to quantify the polarity effect practically, and propose that it should be expressed as a correction factor to be applied to readings with one particular chamber bias. PMID- 15552128 TI - Biomaterial-centered infection: microbial adhesion versus tissue integration. 1987. PMID- 15552129 TI - High-pressure pulsatile lavage causes soft tissue damage. AB - Irrigation and debridement can be a source of iatrogenic injury in open fracture treatment. Although high-pressure pulsatile lavage has been shown to cause considerable damage to bone, little has been written about its effects on soft tissue. The purpose of this study is to quantify and compare the damages on soft tissue caused by high-pressure pulsatile lavage and low-pressure lavage. Forty specimens of fresh ovine muscle were collected and subjected to high-pressure pulsatile lavage or low-pressure lavage, with the delivery orientation being across or in line with the muscle fibers. Ten additional specimens were used as controls. The results show that high-pressure pulsatile lavage causes considerable soft tissue penetration of particulate markers (across, 4.7 mm; in line, 15.6 mm) when compared with low-pressure lavage (across, 0.5 mm; in line, 0.7 mm). Furthermore, all specimens subjected to high-pressure pulsatile lavage showed gross tissue disruption. Fifteen additional samples were obtained to measure cellular death. This was observed at a deeper level for high-pressure pulsatile lavage (median depths: across, 1210 microm; in line, 1335 microm), which was approximately twice that of low-pressure lavage (across, 485 microm; in line, 682 microm). These results show that high-pressure pulsatile lavage penetrates and disrupts soft tissue to a deeper level than low-pressure lavage, causing considerable gross and microscopic tissue disruption. PMID- 15552130 TI - Porcine small intestine submucosa does not show antimicrobial properties. AB - The goal of this study is to examine whether porcine small intestine submucosa (SIS) exhibits antimicrobial properties in a standard in vitro system, without pretreatment with acetic acid or extraction of soluble proteins. Previous animal studies suggest that porcine SIS may have inherent antibiotic properties. Using the guidelines for disk diffusion susceptibility testing by Bauer, 17/64-inch diameter disks made of porcine small intestine submucosa and of gortex were compared with standard antibiotic-impregnated disks against six organisms. The zone of inhibition was measured after 24 hours and minimum bacterial concentrations were determined by serial dilutions of a solution in which porcine small intestine submucosa was allowed to elute for 24 hours. Neither porcine SIS or gortex discs caused inhibition of the growth of any organism. The porcine small intestine submucosa discs showed bacterial growth on top of the discs whereas the gortex did not. Neither the dilutional concentrations of the porcine small intestine submucosa eluent nor the gortex eluent inhibited the growth of any organism. These findings suggest that the porcine small intestine submucosa does not have intrinsic antimicrobial properties. The growth of bacteria on top of the porcine small intestine submucosa suggests that porcine small intestine submucosa itself may provide a favorable environment for the growth of bacteria. More research is necessary to decide what role porcine small intestine submucosa plays in the treatment of infected surgical sites. PMID- 15552131 TI - Gentamicin and vancomycin do not impair experimental fracture healing. AB - The purpose of this study was to determine whether gentamicin or vancomycin impair experimental fracture healing in rats. Forty-one male Wistar rats were divided into three groups, receiving either 1.5 mg/kg of gentamicin intramuscularly twice daily for 3 weeks (n = 15), 25 mg/kg of vancomycin intraperitoneally twice daily for 3 weeks (n = 14), or no treatment (n = 12), starting 7 days after production of closed, nondisplaced, bilateral femoral fractures. The mean 30 minute serum concentrations of gentamicin and vancomycin were 4.5 microg/mL and 35.1 microg/mL, respectively. Biomechanical and radiographic studies were used to evaluate fracture healing. Torsional strength testing of fracture callus exposed to gentamicin and vancomycin was similar (437 and 424 N-mm, respectively) to controls (334 N-mm). Radiographs showed similar healing in control animals (Goldberg score, 2.3) compared with rats treated with gentamicin and vancomycin (Goldberg score, 2.6 in both groups). Results of this study do not show impaired healing of experimental fractures after exposure to gentamicin or vancomycin (such as has been reported after exposure to quinolones). PMID- 15552132 TI - The effect of glycine filler on the elution rate of gentamicin from acrylic bone cement: a pilot study. AB - Elution of antibiotics from acrylic bone cement (polymethylmethacrylate [PMMA]) is dependent on the access of fluid to the depths of the cement that contains the antibiotic. Commercially prepared antibiotic beads that are porous have higher elution rates than hand-mixed, nonporous antibiotic PMMA mixtures. To increase the elution of gentamicin from hand-mixed PMMA, glycine was added as a filler to produce porosity. Elution of gentamicin from the antibiotic PMMA-glycine mixture increased with increasing amounts of glycine. With 3.6 g gentamicin powder and 14 g of crystalline glycine per batch of Palacos PMMA, the elution of gentamicin from the PMMA at 2 days was, similar to the previously documented elution of gentamicin from commercially prepared porous Septopal PMMA beads. With further investigation it may be possible to identify a specific filler and a volume of filler that can be hand mixed in antibiotic PMMA to produce the elution behavior that is needed for specific clinical requirements. PMID- 15552133 TI - Eradication by surfactant irrigation of Staphylococcus aureus from infected complex wounds. AB - Treatment of infected orthopaedic hardware usually requires the removal of the appliance. When the device is removed and immediately replaced, persistent infection frequently complicates this exchange procedure. We modeled the exchange procedure in rats by passing a wire suture through a posterior spinous process and then contaminating the wound with Staphylococcus aureus. We then investigated whether a sequence of surfactant enriched irrigation solutions (Castile soap followed by benzalkonium chloride, sequential surfactant irrigation) had a greater capacity to eradicate Staphylococcus aureus from the experimental wound than did the standard wound irrigant, normal saline. When we left the wire in place through the 2-week course of the study, sequential surfactant irrigation showed only a modest advantage over normal saline (staphylococci recovered from 39% versus 58% of wound cultures respectively). Simple removal of the wire 24 hours after implantation and bacterial contamination prevented wound infection in most animals (with the wire removed, 38% of the animals remained infected versus 85% with the wire left in place), without regard to the irrigation solution. Alternatively, when we removed the wire after 24 hours, irrigated the wound, and then placed a fresh wire back into the wound, sequential surfactant irrigation showed a significant advantage over NS (54% of the animals irrigated with sequential surfactants remained infected versus 100% of the animals irrigated with normal saline). Our findings confirm the importance of a contaminated medical device for promoting foreign body infection; our findings also show that sequential surfactant irrigation has therapeutic value in a rat model of orthopaedic device infection; this irrigation protocol should be studied further as a potential agent for the treatment of infected orthopaedic wounds. PMID- 15552134 TI - Successful treatment of total hip and knee infection with articulating antibiotic components: a modified treatment method. AB - There are many problems associated with the use of articulating antibiotic cement spacer blocks and articulating components in the two-stage treatment of total hip replacement and total knee replacement infections. These include loss of motion during and after treatment, bone loss, generation of cement debris, inadequate dosing of cement with the appropriate antibiotic, and biologic failure. Forty four patients with 54 consecutive periprosthetic hip and knee infections (31 septic total knee arthroplasties and 23 septic total hip arthroplasties) had treatment with a modified two-stage reimplantation protocol using articulating components made of antibiotic-cement-only prosthetic components and antibiotic cement-covered prosthetic components between January 1995 and May 2002. Second stage revision, after six weeks of parenteral antibiotics, was completed an average of 84 days after the first stage. A minimum two-year followup after final treatment is evaluated. One of the 23 total hip replacement infections persisted or recurred with the original organism(s) after treatment (95.7% success) as did two of the 31 total knee replacement infections (93.5% success). Combined success rate was 94.4%. This modified treatment method incorporates early range of motion during first-stage treatment with articulating components that provide local high dose elution of broad-spectrum antibiotics, provides the flexibility of customizing the antibiotic cement components with culture-directed antibiotics, and results in a high biologic success rate. PMID- 15552135 TI - Systemic safety of high-dose antibiotic-loaded cement spacers after resection of an infected total knee arthroplasty. AB - The purpose of this study was to assess the systemic safety and potential adverse effects of using a high-dose antibiotic-impregnated cement spacer after resection arthroplasty of an infected total knee replacement. Between October 2000 and December 2002, 36 knees (34 patients) had a resection arthroplasty of an infected total knee prosthesis with placement of a high-dose antibiotic impregnated cement spacer. There were 24 men and 10 women with a mean age of 66.5 years (range, 48 84 years). All spacers placed contained an average of 3.4 batches of cement with an average total dose of 10.5 g of vancomycin (range, 3-16 g) and 12.5 g of gentamicin (range, 3.6-19.2 g). All patients were followed up post-operatively until reimplantation for evidence of renal failure. The preoperative creatinine ranged from 0.7 to 1.8 mg/dL. All patients were concomitantly treated with 6 weeks of intravenous organism-specific antibiotics. One patient with normal preoperative renal function (Cr 0.7 mg/dL) had a perioperative 1-day transient rise in serum creatinine (1.7 mg/dL) postoperatively that subsequently normalized. No patients showed any clinical evidence of acute renal insufficiency, failure, or other systemic side effects of the antibiotics. Treatment of patients with an infected total knee arthroplasty with high-dose vancomycin and gentamicin antibiotic spacers seems to be clinically safe. PMID- 15552136 TI - Complications and results of arthroplasty for salvage of failed treatment of malignant pathologic fractures of the hip. AB - The purpose of this study was to evaluate the results and complications of hip arthroplasty done for salvage of failed treatment of pathologic proximal femoral fractures secondary to malignancy. Between 1980 and 2000, 42 patients with a mean age of 63 years were treated with hip arthroplasty to salvage failed treatment of a pathologic proximal femoral fracture. Total hip arthroplasty was done in 16 patients (3 uncemented, 2 hybrid, 11 cemented), and bipolar hemiarthroplasty in 26 (2 uncemented, 24 cemented). A modular, proximal femoral replacement construct was used in 15 patients. Patients were followed a mean of 5.8 years (range, 15 days-20 years). Four hips required reoperation, all for deep infection. Harris Hip score improved from an average of 42 points (range, 17-76 points) preoperatively to an average of 83 points (range, 52-100 points) postoperatively. Most recent radiographs showed femoral component loosening in only one patient. Implant survivorship free of revision for any reason at 5 years was 90% (range, 65-96%) and free of revision for aseptic failure or radiographic failure was 97% (range, 64-99%). Hip arthroplasty is an effective treatment for salvage of failed treatment of pathologic proximal femoral fractures. Modular proximal femoral replacements were often required. The most concerning complication was deep prosthetic infection, which occurred in nearly 10% of this patient population, and in 21% of patients with prior irradiation. PMID- 15552137 TI - Infected fractures of the distal tibial metaphysis and plafond: achievement of limb salvage with free muscle flaps, bone grafting, and ankle fusion. AB - Infection after fractures of the tibial plafond is a challenging problem that may even result in amputation. The current study evaluates a limb salvage protocol and the associated long term functional outcome in 6 patients (mean age 46 years) who were treated for infection after a fracture of the distal tibial metaphysis and plafond. Our limb salvage protocol included 3 stages: 1) radical debridement and stabilization of the ankle with a bridging external fixator, 2) soft tissue coverage with free muscle flaps, and 3) ankle fusion using iliac crest bone graft for filling the existing defects measuring 4.2 cm on average. At a mean followup of 5.5 years (range, 2-10.5 years), limb-salvage and eradication of infection was accomplished in all extremities. Fusion of the ankle joint was achieved in all patients, with one patient requiring a supplemental bone grafting procedure for delayed healing of the fusion site. All patients are able to walk without assistive devices and five of six patients are pain free. Limb salvage with free muscle flaps, bone grafting, and ankle fusion is a viable option for the treatment of infected tibial metaphysis and plafond fractures. PMID- 15552138 TI - Wound complications after open Achilles tendon repair: an analysis of risk factors. AB - Operative treatment of Achilles tendon ruptures has the risk of wound complications. The purpose of this study is to determine the risk ratio for specific risk factors associated with wound related complications in patients with operatively treated Achilles tendon ruptures. Between 1978 and 2001, 167 open Achilles tendon repairs were done at our institution. Clinical data were retrospectively reviewed. Tobacco use, diabetes, age, gender, timing of surgery, body mass index and steroid use were evaluated as potential risk factors for wound healing complications. One patient was lost to follow-up and two patients had nonsimultaneous ruptures and only the first repair was included; the remaining patients were followed up until complete wound healing. There were 17 wound complications in 164 patients (10.4%). Significant risk factors for development of wound complications included tobacco use (p < 0.0001), steroid use (p = 0.0005), and female sex (p = 0.0400). For those patients who had one or more of the following risk factors: diabetes, tobacco use, or steroid use; eight of 19 (42.1%) had a complication, compared with nine of 145 (6.2%) for those without risk factors present (p < 0.0001). Surgeons doing open Achilles tendon repairs should be cognizant of the specific risk factors identifled in this study, because they might impact decision making with regard to operative versus nonoperative treatment. PMID- 15552139 TI - Successful treatment of chronic bone and joint infections with oral linezolid. AB - Linezolid is an attractive alternative for the treatment of chronic bone and joint infections because it is active against common pathogens including methicillin-resistant staphylococci and vancomycin-resistant enterococci and because its oral formulation is convenient for long-term administration. To evaluate the ability of linezolid to produce long-term remission, we prospectively monitored 11 consecutive adult patients who received linezolid for osteomyelitis (n = 9) or prosthetic joint infection (n = 2). Linezolid 600 mg was administered orally twice daily for a mean of 10 weeks (range, 6 to 19 weeks). Pathogens were methicillin-resistant Staphylococcus aureus (n = 5), methicillin resistant coagulase-negative staphylococci (n = 4), vancomycin-resistant Enterococcus faecium (n = 1), and vancomycin-sensitive Enterococcus faecalis (n = 1). After a mean followup of 27 months (range, 17 to 41 months), all 11 patients had remission by clinical, laboratory, and radiographic criteria. During week 8 of linezolid treatment, one patient developed a gram-negative superinfection, which resolved with appropriate therapy. During week 6 of linezolid treatment, one patient developed mild thrombocytopenia and another patient developed mild anemia. Both episodes of myelosuppression were reversible within 10 days after completing the planned 6-week courses of linezolid. We recommend weekly complete blood counts to detect hematologic abnormalities. We conclude that oral linezolid seems to be a useful and convenient alternative for the treatment of bone and joint infections. PMID- 15552140 TI - The current status of material used for depot delivery of drugs. AB - The ideal local antibiotic delivery system has not been created. Antibiotic-laden bone cement has become the gold standard in the treatment of infected orthopaedic implants and there are confirmatory laboratory and clinical data that support the use of these materials. Heat-stable antibiotics elute from antibiotic-laden bone cement and do not have a notable influence on the compressive strengths of bone cement if the antibiotics are used in appropriate amounts. If the proper antibiotic is chosen, placed in the appropriately porous materials in sufficient amounts, and implanted in bone, antibiotic levels in the surrounding bone are many times greater than can be achieved by safe systemic antibiotic doses. Although the materials that have been manufactured commercially have been used for over 30 years in Europe, until recently, they have not been available in the United States. Currently, there are five antibiotic-laden bone cement composites that have been approved by the FDA and that are available for clinical use. Studies are being done to search for biodegradable implants preferable to antibiotic-laden bone cement; however, these studies and the materials are still in early stages and development. Currently, there are no FDA-approved biodegradable materials available for use to treat infected orthopaedic implants. As new materials become available and their elution characteristics are recorded, it is important for surgeons to understand how the data were collected so they can have a clear understanding of the elution characteristics of the material used and how the material acts in different environments. Even with extensive historic, clinical, and research data that prove the effectiveness of antibiotic laden cement, the ideal drug delivery system is neither agreed on nor available. PMID- 15552141 TI - Practical applications of antibiotic-loaded bone cement for treatment of infected joint replacements. AB - The use of antibiotic-loaded bone cement is an accepted treatment method for infected joint arthroplasties. It is helpful to separate the use of antibiotic loaded bone cement as a method of prophylaxis as compared with the treatment of an established infection. A low dose of antibiotic-loaded bone cement (< or = 1 g of antibiotic per batch of cement) should be used for prophylaxis, and high-dose antibiotic-loaded bone cement (> 1 g antibiotic per batch of cement) is indicated for treatment. The only commercially available antibiotic-loaded bone cement products are low dose, with the use of tobramycin or gentamicin as an antibiotic selection. High-dose antibiotic-loaded bone cement requires hand mixing by the surgeon to facilitate the use of high dosages and choices of multiple antibiotics. Treatment of infected hip and knee arthroplasties with high-dose antibiotic-loaded bone cement is aided by the use of spacers of various shapes and sizes. These spacers, whether they are static or articulating (mobile), are meant to provide local delivery of antibiotics, stabilization of soft tissues, facilitation of an easier reimplantation, and improved clinical outcomes. PMID- 15552142 TI - Local antibiotic therapy in the treatment of open fractures and osteomyelitis. AB - Local antibiotic therapy is part of the treatment protocol of open fractures and osteomyelitis. However, there are limitations in the literature evaluating the efficacy of the technique and controversies remain. Polymethylmethacrylate cement commonly is used as a delivery vehicle, but bioabsorbable vehicles are being investigated. The vehicle is impregnated with antibiotic(s) active against the suspected pathogens, usually an aminoglycoside and/or vancomycin. Elution depends on the surface area, the characteristics and permeability of the delivery vehicle, type and concentration of the antibiotic(s) used, presence of fluid, rate of fluid turnover, and time postimplantation. Local antibiotic therapy is a safe technique resulting in high local concentration of antibiotics with minimal systemic levels. Local antibiotics effectively have controlled infection in animal models and, despite limitations of the existing literature, seem to be useful in the clinical setting. The length of implantation and the need for removal of the delivery vehicle remain controversial. Nonabsorbable vehicles may eliminate the need for reoperation and removal. PMID- 15552143 TI - Two-stage reimplantation of total joint infections: a comparison of resistant and non-resistant organisms. AB - The prevalence of resistant organisms has increased throughout the last decade. In order to determine best treatment strategies, studies on reimplantation must address this population. The purpose of this study is to compare the results of delayed reconstruction for total joint infections caused by methicillin-sensitive and methicillin-resistant organisms. This investigation included 46 patients (21 men, 25 women) with an average age 67 years (range, 36-86 years) who had a two staged reimplantation after periprosthetic total joint infection. Thirty-seven patients cultured methicillin-sensitive organisms and nine cultured methicillin resistant organisms. Treatment included resection, aggressive debridement, 6 weeks of parenteral antibiotics, and subsequent debridement and reimplantation. Treatment was considered successful if the patient was disease free at last followup (average 5 years, range 2-10 years). The efficacy of two-stage reconstruction was similar for infections with sensitive and resistant bacteria. Only 3 of 46 patients failed delayed reconstruction requiring further treatment. Failure rates with regards to infections with sensitive bacteria were 5.4% (2 of 37 infections) and resistant bacteria 11.1% (1 of 9 infections), which was not significant. The current study shows clinical success of two-stage reimplantation for patients with methicillin-resistant bacteria complicating total joint arthroplasty. PMID- 15552144 TI - Alternative materials to acrylic bone cement for delivery of depot antibiotics in orthopaedic infections. AB - Acrylic bone cement has considerable laboratory and clinical data validating it as a delivery material for depot administration of antibiotics. However, an alternate material that does not require a secondary procedure for removal is desired. Many biodegradable materials have been evaluated as alternatives including protein-based materials (collagen, fibrin, thrombin, clotted blood), bone-graft, bone-graft substitutes and extenders (hydroxyapatite, beta-tricalcium phosphate, calcium sulfate, bioglass), and synthetic polymers (polyhanhydride, polylactide, polyglycolide, polyhydroxybutyrate-co-hydroxyvalerate, polyhydroxyalkanoate). Various forms and combinations of these materials have been investigated worldwide, characterizing their elution properties and performance in treating osteomyelitis in animal models. Many of these have had limited clinical evaluation. Outside the United States, some of these materials are used clinically. In the United States, none have been approved. None are commercially available for clinical use. Morselized cancellous bone and calcium sulfate are the two materials that have been used clinically in the United States on a physician-prescribed, hand-mixed, basis. Considering the limited clinical data that currently are available, the use of these materials still is experimental. Clinical application should be cautious, limiting the total antibiotic load. Until definitive data are available, a prudent dose would be no higher than one that would have acceptable toxicity risk if administered intravenously over 24 hours. PMID- 15552145 TI - Shoulder scoring scales for the evaluation of rotator cuff repair. AB - Various assessment tools have been proposed for evaluation of shoulder function. Analyses of comparability, validity, and reliability among shoulder assessment tools are lacking. The purpose of our investigation was to compare the results of three commonly used shoulder assessment tools in the evaluation of a specific shoulder condition. Seventy-two full-thickness rotator cuff tears were treated with an open rotator cuff repair from 1986-1993. The average age of the patients at surgery was 58 years (range, 24-92 years). The median duration of followup at the time of evaluation for this study was 55 months (range, 24-102 months; standard deviation, 22 months). Correlation among the results of the UCLA, Constant-Murley, and the Simple Shoulder Test scales was fair (range, 0.66-0.76). More importantly, the three systems were evaluated for their ability to accurately predict improved motion, strength, and patient satisfaction. The Simple Shoulder Test and the Constant-Murley scales showed the highest positive predictive values. The Simple Shoulder Test is a patient-based self-assessment device that easily can be incorporated into a busy clinical practice, providing outcome data comparable with complex evaluation systems when evaluating the results of rotator cuff repair. PMID- 15552146 TI - Ultrasonographic stress test of the metacarpophalangeal joint of the thumb. AB - The purpose of this study was to evaluate functional ultrasound as a tool for detecting an ulnar collateral ligament injury of the thumb. The feasability of using ultrasound for imaging the thumb joint space was tested in a pilot study, using ultrasound and dissection in 14 cadaveric hand specimens. To test this method clinically, both metacarpophalangeal joints of the thumb in 461 healthy volunteers were examined using ultrasound (11 MHz) under radial stress. The distance between the innominate tubercle of the first metacarpal head to the proximal phalanx was measured. The mean distance between the first metacarpal head and the proximal phalanx (n = 461) was 4.5 mm (standard deviation, 0.65 mm) on the right side and 4.6 mm (standard deviation, 0.61 mm) on the left side. These data were compared with data of 25 patients with an operative diagnosis of rupture of the ulnar collateral ligament of the thumb. The difference in joint space between the injured and uninjured sides was 2.25 mm (standard deviation, 0.46 mm). The metacarpophalangeal joint space can be reproducibly detected on high-frequency ultrasound. An increased gap seen on ultrasound is indicative of a rupture of the ulnar collateral ligament of the thumb. PMID- 15552147 TI - Dose-responsiveness of electrophysiologic change in a new model of acute carpal tunnel syndrome. AB - This study was done to determine the dose-responsiveness during the development of acute pressure-induced median neuropathy in rabbits and to develop a new animal model of acute carpal tunnel syndrome. Twenty-three rabbits were used. Carpal tunnel syndrome was induced by infusing a controlled saline solution into the carpal tunnel of rabbits using a 21-gauge needle under general anesthesia to elevate the carpal tunnel pressure. The changes in the amplitude and latency of the compound muscle action potential obtained by abductor pollicis recording were observed after a complete conduction block, and after the release of pressure at various pressure levels. Pressures greater than 30 mm Hg applied to the carpal tunnel caused a remarkable electrophysiologic change. A higher pressure level resulted in a shorter time required for a complete conduction block and also for recovery after the release of pressure. Complete conduction block occurred between 40 and 50 minutes at a pressure of 100 mm Hg. The degree of recovery after a conduction block was related inversely to the pressure-time integrals. Our animal model reflects the pathophysiology of acute carpal tunnel syndrome and shows a regular dose-responsiveness during the development of acute pressure induced neuropathy. PMID- 15552148 TI - MRI can identify high intensity bands around implants that correspond to radiolucent lines on radiographs: an ex vivo study of sheep acetabula. AB - We investigated the use of magnetic resonance imaging in categorizing the tissue interface between bone and bone cement after total hip replacement. In an ex vivo sheep model, we compared correlations between the thicknesses of radiolucent lines measured from plain radiographs and corresponding high signal intensity bands in magnetic resonance imaging scans. Correlations obtained for the three main Gruen Zones were in the range between r2 = 0.58 (superior zone), and r2 = 0.86 (inferior zone). In two specimens magnetic resonance imaging was able to detect high signal intensity bands that were not visible on the radiographs. The average thickness of the high signal intensity bands measured on the magnetic resonance imaging scans ranged from 14.6% (Zone 2) to 39.9% (Zone 3) larger than the corresponding radiolucent lines measured on the plain radiographs. Magnetic resonance imaging has the advantage over current techniques in that it is noninvasive and does not use ionizing radiation. If problems associated with metal artifacts can be reduced, these potential advantages may make magnetic resonance imaging the preferred method for longitudinal studies assessing osteointegration and analysis of fibrous membranes. PMID- 15552149 TI - Polymerase chain reaction can detect bacterial DNA in aseptically loose total hip arthroplasties. AB - Identifying low-grade infection in failed total hip arthroplasties is an important but difficult task. This study investigated the ability of the polymerase chain reaction to identify low-grade infection during revision of total hip arthroplasties that failed from aseptic causes. One hundred thirteen specimens from 31 total hip arthroplasties revised for aseptic loosening were compared with 105 control specimens from 28 primary total hip arthroplasties. All surgeries were done in laminar flow operating rooms. No primary or revision specimen had positive microbiologic cultures. No revision specimen had histologic evidence suggestive of infection. Using the polymerase chain reaction with a detection threshold of 10 organisms per cubic centimeter of specimen, bacterial DNA was identified in 39 of 85 revision tissue specimens (46%) compared with 18 of 84 primary tissue specimens (21.4%). Bacterial DNA was identified in the synovial fluid of three specimens taken from 28 revision total hip arthroplasties (10.7%) and in two specimens taken from 21 primary total hip arthroplasties (9.5%). As multiple specimens were sent for each hip, a maximum of 16 of 31 revision total hip arthroplasties (52%) and eight of 28 primary total hip arthroplasties (29%) were considered to be infected. Bacterial DNA can be found in many specimens obtained from revised total hip arthroplasties considered to be aseptically loose. Because bacterial DNA identified at primary total hip arthroplasty was assumed to be attributable to contamination rather than present in healthy tissues, the overall specimen contamination rate of 19% and case contamination rate of 29% indicate that the polymerase chain reaction has poor specificity at this sensitivity level for diagnosing infection in revision total hip arthroplasty. PMID- 15552150 TI - Osteolysis: medical and surgical approaches. AB - Osteolysis is defined as the process of progressive destruction of periprosthetic bony tissue, characterized on serial radiographs as progressive radiolucent lines and/or cavitation at the implant-bone or cement-bone interface. Without proper treatment, osteolysis may progress to aseptic loosening and catastrophic failure of the implant. Osteolysis is a multifactorial process stemming from host, prosthesis, and surgical factors. Billions of submicron wear particles are generated at material interfaces that cause osteolysis. Wear particles are dispersed along the effective joint space, into bone, and throughout the adjacent soft tissues. Initially, patients may have no clinical symptoms despite radiographic evidence of osteolysis and can remain completely asymptomatic, even in the presence of substantial bone loss. Symptoms usually do not occur until the bone loss has reached the point of causing implant loosening, implant failure (fracture), or periprosthetic fracture. Radiographic images and standard examination provide limited information and almost always underestimate the amount of bone loss. Three-dimensional computed tomography is more sensitive and can monitor the volume of the lesion, however the trade-offs are its high cost and radiation exposure. The timing for treatment of periprosthetic osteolysis is controversial and the medical decision-making is debatable. Medical treatment seems to be attractive but there is no proven or approved drug therapy to prevent or inhibit periprosthetic osteolysis. We reviewed the medical and surgical options that are available for treatment of periprosthetic osteolysis. PMID- 15552151 TI - Titanium and cobalt-chrome cementless femoral stems of identical shape produce equal results. AB - This study of proximal porous-coated stems of identical shape, but with two types of stem materials (Ti alloy or Co-Cr alloy), was done to determine the differences, if any, in these stems in the following parameters: clinical and functional outcomes; the prevalence of thigh pain; the stem alignment and the stem-canal fill; cup position; the degree of periprosthetic bone loss; the prevalence of polyethylene liner wear and osteolysis; the incidence of aseptic loosening of acetabular and femoral components; and complication rates. In the Ti alloy group, there were 60 patients (70 hips) followed up for a mean of 6.4 years (range, 5-7 years) with a mean age at surgery of 45.3 years and a mean weight of 65.7 kg. In the Co-Cr alloy group, there were 60 patients (70 hips) followed up for a mean of 6.6 years (range, 5-7 years) with a mean age at surgery of 45.4 years and a mean weight of 66.8 kg. Harris hip scores did not differ and the prevalence of transient pain in the thigh was 5% with both stems. No acetabular or femoral component had aseptic loosening in either group. Loss of bone density in the calcar region was more severe in the Co-Cr group. The mean polyethylene linear wear per year was 0.15 mm (SD, 0.015) in the Ti group and 0.14 mm (SD, 0.013) in the Co-Cr group. No acetabular or femoral osteolysis was seen in any patient. The clinical and radiographic results of this study did not show any significant differences between the Ti and Co-Cr alloy femoral components; however, the Ti alloy femoral component retained greater periprosthetic bone as compared with a Co-Cr alloy femoral component. PMID- 15552152 TI - Primary uncemented Harris-Galante acetabular components in patients 50 years old or younger: results at 10 to 12 years. AB - Seventy-three patients (84 hips) 50 years or younger were treated with primary total hip arthroplasty using the Harris-Galante uncemented acetabular component. At a median followup of 10 years, there were seven acetabular metal shell revisions: five for osteolysis and one each for aseptic loosening and dislocation. There were 10 polyethylene exchanges without metal acetabular shell removal. None of the unrevised acetabular components was radiographically loose, but eight had pelvic osteolysis. The rate of 10-year survival without revision of the acetabular metal shell was 87.9% (95% confidence interval, 80.1%-95.7%). The same rate of survival without revision for aseptic acetabular component loosening was 97.3% (95% confidence interval, 92.9%-100%). Revision for aseptic acetabular loosening or polyethylene liner exchange or both was 84.5% (95% confidence interval, 75.8%-93.4%). PMID- 15552153 TI - Total femoral arthroplasty for salvage of end-stage prosthetic disease. AB - The complications and outcomes in patients who had total femoral arthroplasty for salvage of a severely compromised femur were studied. The clinical scenarios included numerous revision total hip or knee arthroplasties, failed periprosthetic fractures, or recurrent infection treated with multiple radical debridement surgeries. Fifty-nine patients (average age, 73.7 years) were identified. At an average 4.8 years followup, adequate pain relief was achieved and Harris hip pain scores averaged 33.8 of 44 points, and knee pain scores averaged 42.8 of 50 points. Good function was achieved with 98% of patients able to ambulate and 43% using no assistive device or a cane only. There were 18 complications or subsequent surgeries. Infection and dislocation occurred in eight patients and seven patients, respectively. Total femoral arthroplasty for salvage of a severely compromised femur provides acceptable results even in the most difficult of cases. PMID- 15552154 TI - An intraoperative pressure-measuring device used in total knee arthroplasties and its kinematics correlations. AB - Fluoroscopic and retrieval analyses of knee implants show considerable variability even for the same implant design, and implicate the possible importance of surgical technique and compartment pressure balance in total knee arthroplasties. This study was done to correlate intraoperative computer-assessed compartment pressure measurements with postoperative kinematics to explain these variations. Thirty-eight patients had posterior cruciate-sacrificing low-contact stress total knee arthroplasties using a balanced gap technique. At trial reduction, an instrumented tibial insert designed to record the magnitude, location, and dynamic imprint of the pressures in the medial and lateral compartments was placed into the knee. Pressures were recorded electronically for a range of motion from 0 degrees - 120 degrees. Sixteen of the 38 patients agreed to do successive weightbearing deep knee bends under fluoroscopic surveillance. Only three of the 16 patients had condylar lift-off, but all experienced lift-off at a single flexion angle. In the three patients who had condylar lift-off, a compartment pressure imbalance, as measured by the intraoperative pressure sensor, occurred at the same flexion angle of lift-off. These data suggest that although a given implant design may have inherent kinematic tendencies, surgical technique and compartment pressure balance significantly impact kinematic performance. PMID- 15552155 TI - Clinical significance of gelatinases in septic arthritis of native and replaced knees. AB - We hypothesized that more gelatinases appear in effusions of septic arthritis than aseptic arthritis. This study examined the laboratory variables of inflammation and the levels of gelatinase A and B (matrix metalloproteinases-2 and -9) in 75 effusions from the knees of 37 patients with inflammatory arthritis and compared them with effusions of septic and aseptic arthritis. Gelatin zymography revealed that the levels of the latent matrix metalloproteinase-9 were higher in 24 effusions of septic arthritis than in 51 effusions of aseptic arthritis. The latent matrix metalloproteinase-9 levels of septic arthritis also correlated with the neutrophil counts in effusions. Significantly more activated matrix metalloproteinases-2 and -9 appeared in effusions of septic arthritis in native and replaced knees than in effusions of aseptic arthritis. A high matrix metalloproteinase-9 level and the appearance of activated matrix metalloproteinases-2 and -9 may distinguish septic from aseptic arthritis, even in cases with a low neutrophil count in the replaced knee. Joint aspiration may not only reduce the bacteria counts, endotoxins, and proinflammatory cytokines, but also decrease the amount of matrix metalloproteinases in effusions that attack the extracellular matrix of native and artificial joints. PMID- 15552156 TI - Chondrosarcoma of the chest wall: a retrospective clinical analysis. AB - Primary chondrosarcoma of the chest wall is uncommon, and reports of treatment from one institution are limited. Treatment of this lesion is difficult because of the anatomic location, which is in close proximity to surrounding neurovascular and visceral structures. The purpose of this study was to review the outcome of surgical resection of chondrosarcoma of the chest wall, and to delineate the prognostic factors related to local recurrence and survival. Twenty four patients with chondrosarcoma of the chest wall were treated at our institution between 1986 and 2000. There were 14 males and 10 females, with an average age of 54 years (range, 11-76 years). Patients were observed for a minimum of 3 years or until death. The median followup was 71 months. The anatomic locations of chondrosarcoma of the chest wall were the rib in 16 patients, the ribs and sternum in two patients, the ribs and spine in three patients, and the sternum only in three patients. Histologically, 17 patients had Grade 1 disease and seven patients had Grade 2 disease. At followup, 17 patients were alive without disease, two were alive with disease, two died without disease, and three died secondary to progressive disease. The overall survival estimate at 5 years was 92%. The recurrence rate for patients with adequate surgical margins was 10%, compared with 75% for patients with inadequate margins. The 5-year survival rate for patients with adequate surgical margins was 100%, compared with 50% in patients with inadequate surgical margins. An inadequate margin of resection was associated with a significantly worse overall survival and a higher chance of having local recurrence develop. PMID- 15552157 TI - The use of cement in osteoarticular allografts for proximal humeral bone tumors. AB - In a proximal humerus resection for a bone tumor, the use of an osteoarticular allograft is considered the best restoration of shoulder function. We retrospectively reviewed the outcomes of 31 patients who had an intraarticular resection of the proximal humerus for a bone tumor. Twenty-three of the allografts were filled with cement. The average followup was 5.3 years. Of the 31 patients with more than 24 months followup, seven had revision surgery or removal of the allograft. Kaplan-Meier analysis showed that the probability of survival of the reconstruction was 78% at 5 years. Fracture was the main complication in 11 patients (37%) of whom seven were in the noncemented group. Four of these patients had successful surgery for conversion to an allograft-prosthetic composite, whereas one patient had a new allograft. Allografts that were filled with cement had four fractures (18%); three were subchondral fractures discovered by routine CT scans. None of these patients had pain or needed revision surgery. Osteochondral allograft in proximal humerus replacement is a reliable reconstructive technique if the allograft is augmented by filling the intramedullary space with cement. Moreover, cement augmented allografts are less expensive and technically easier than allograft-prosthetic composites. PMID- 15552158 TI - Phalangeal osteoid osteomas in the hand: a diagnostic problem. AB - Osteoid osteomas in the phalanges of the hand often present with atypical clinical and radiologic characteristics. These unusual presentations may cause a delay in diagnosis and treatment. In this study, seven cases (seven patients) of phalangeal osteoid osteomas in the hand were examined, six of which were misdiagnosed for long periods of time. These seven cases illustrate unusual presentations, including absence of reactive bone, monoarticular arthritis, clubbing, macrodactyly, painless swelling, and absence of bony lysis. Phalangeal osteoid osteomas also have unusual histologic features. Recognition of these unusual clinical, radiographic, and histologic features may prevent misdiagnosis and lead to prompt definitive therapy. PMID- 15552159 TI - Intertrochanteric versus subcapital osteotomy in slipped capital femoral epiphysis. AB - We reviewed 15 flexion intertrochanteric osteotomies and 11 subcapital osteotomies done for chronic, severe, stable slipped capital femoral epiphysis at one institution with a minimum 2 years followup. The goal was to determine which procedure achieved better deformity correction and which procedure was safer. Radiographically, the two groups were equivalent in compensating for epiphyseal slip. Flexion intertrochanteric osteotomy was more effective in restoring proximal femoral anatomy, as determined by articulotrochanteric distance and trochanter-center of head distance. The incidence of complications, including osteonecrosis and chondrolysis, was low for both groups, but the reoperation rate was greater in the subcapital osteotomy group than in the flexion intertrochanteric group. Flexion intertrochanteric osteotomy seems to be an effective, safe, and reproducible realignment osteotomy for treatment of chronic, severe, stable slipped capital femoral epiphysis. PMID- 15552160 TI - Early loss of hip containment in a child with dysplasia epiphysealis hemimelica. AB - Progressive loss of hip containment attributable to dysplasia epiphysealis hemimelica of the right proximal femur and macrodactyly of the right second toe was diagnosed and monitored by radiographs and magnetic resonance imaging in a 7 year-old boy. The patient had early surgical correction including shortening osteotomy of the hypertrophic toe and partial resection of the involved superolateral femoral head-neck junction combined with a Pemberton-like acetabuloplasty. This treatment restored containment and function. At 4 years followup, the femoral head remained contained and the patient was participating fully in the activities of daily life for his age-group. A review of the literature suggests monitoring the hip with magnetic resonance imaging may allow early identification of a hip at risk for loss of containment. This treatment may save the hip from severe deformity and early secondary osteoarthritis. PMID- 15552161 TI - Molecular regulation of osteoblasts for tissue engineered bone repair. AB - The use of biodegradable polymers in medicine and biomedical research is increasing. A key growth area has been the use of these materials in tissue engineering, especially for guided regeneration of bone and cartilage. Our interest has been in determining the mechanisms by which cellular attachment and growth occurs on these materials. In the current study, we examined human osteoblast cell adhesion, growth, and morphologic changes on polymeric scaffolds composed of polylactic-co-glycolic acid and polylactic acid materials. We examined these characteristics in association with measurements of levels of key adhesion integrin receptors in the presence and absence of antibodies against alpha2, alpha3, alpha4, alpha5, alpha6, and beta1 subunits, and the adhesion ligand peptides RGD (Arg-Gly-Asp) and RGE (Arg-Gly-Ser). At 2 hours, results showed initial cell adhesion was considerably decreased on polylactic-co-glycolic acid and polylactic acid in the presence of the alpha2 and beta1, antibodies with a 70% adhesion rate difference observed among the groups evaluated. Higher levels of inhibition were observed on polylactic-co-glycolic acid relative to polylactic acid, which may be correlated to a higher number of cells being able to interact with the surface initially. The presence of known competitive peptide (RGD) at 2 hours, revealed its ability to block cellular adhesion to these matrices relative to the control and noncompetitive peptide RGE on polylactic-co-glycolic acid matrices. Overall adhesion rate was affected by the presence of the integrin antibodies to the alpha2, alpha3, alpha4, alpha5, alpha6, and beta1 subunits with highest differences among polylactic-co-glycolic acid relative to its control, therefore suggesting that initial osteoblastic cell adhesion to commonly used biomaterials is regulated through integrin binding. PMID- 15552162 TI - Mini-pig fresh osteochondral allografts deteriorate after 1 week of cold storage. AB - As a well-defined animal transplantation model, the mini-pig potentially is well suited for large animal studies of fresh osteochondral allograft transplantation. This study was done to determine the histologic characteristics and function of proteoglycan synthesis of mini-pig articular cartilage after refrigeration in basal media for as much as 6 weeks. Osteochondral sections of 10 mini-pig knees were refrigerated in various media at 4 degrees C for 1 to 42 days after slaughter. Four hundred twenty samples were evaluated by 35S uptake and 260 samples by histologic evaluations. Proteoglycan synthesis declined by 7 days to 21% of the level measured on Day 1 and was undetectable at 42 days. Histologic evaluation revealed progressive degeneration. Mankin scores rose from 3.69 +/- 0.27 on Day 1 to 6.40 +/- 0.18 on Day 7, and logarithmically increased to 10.83 +/- 0.07 on Day 42. These results indicate that the metabolic characteristics of porcine articular cartilage were not retained after refrigeration in basal media for 7 days. Optimum cold storage of porcine osteochondral allografts for cartilage transplantation research may be less than 7 days. Because osteochondral grafts for clinical use currently are stored for greater than 7 days, similar studies of the viability of human articular cartilage are needed. PMID- 15552163 TI - Lack of efficacy of low-intensity pulsed ultrasound on prevention of postmenopausal bone loss evaluated at the distal radius in older Chinese women. AB - A within-subject, randomized, and prospective intervention trial was done to evaluate the potential effect of low-intensity pulsed ultrasound on prevention of postmenopausal bone loss. Twenty healthy postmenopausal women between the ages of 51-81 years met the inclusion criteria. The treatment hand was randomly selected, and the contralateral site served as a control. Integral and trabecular bone mineral density were measured using highly precise multilayer peripheral quantitative computed tomography at the bilateral distal radius at baseline, 3 months after daily low-intensity pulsed ultrasound treatment, and 3 months after discontinuing treatment. Results showed that the rate of bone change (trabecular bone mineral density and integral bone mineral density) did not significantly differ between the site treated with low-intensity pulsed ultrasound and the contralateral control at either followup. Also, during the followup, bone mineral density did not change significantly in the contralateral control site. This was the first prospective and randomized study to show that low-intensity pulsed ultrasound at the current regime did not have significant effects on intact bone for prevention of postmenopausal bone loss in the distal radii of older Chinese women. PMID- 15552164 TI - Fluorescent microspheres are reliable for serial bone blood flow measurements. AB - The fluorescent microsphere method is one of the current techniques to determine regional blood flow in various organs. The purpose of this study was to examine the suitability of fluorescent microspheres for serial measurement of regional bone blood flow. Six anesthetized female New Zealand rabbits received five left ventricular injections of fluorescent microspheres in 20-minute intervals. To test the precision of the measurement two types of fluorescent microspheres were injected simultaneously at the first and last injections. Blood flow was calculated in the kidneys, lungs, brain, femurs, and tibias after measuring the fluorescence intensity in each reference blood and tissue sample. Comparison of blood-flow values obtained by simultaneously injected microspheres showed an excellent correlation and a minimal percentage difference at the first and last injections, indicating valid measurements of regional bone blood flow. No significant differences were observed when comparing blood flow in the corresponding regions of bones on the right side and left side. Mean blood flow in the femur and tibia significantly increased at the fourth injection whereas flow distribution within the femur and tibia essentially remained unchanged throughout the experiment. Comparison of blood flow values obtained by simultaneously injected microspheres showed moderate agreement for the kidneys and lungs at the last injections. Because this finding might be attributable to disturbances of microcirculation caused by accumulation of spheres in high-flow organs, the increase in regional bone blood flow observed in our experiments has to be interpreted carefully. This study showed that bone blood flow can be determined reliably in anesthetized rabbits by as many as three serial injections of fluorescent microspheres. PMID- 15552165 TI - Systematic reviews of nonrandomized clinical studies in the orthopaedic literature. AB - We systematically reviewed systematic reviews of surgical orthopaedic interventions published between 1996 and 2001 to document when and how nonrandomized studies were included. From more than 10,000 citations examined in various electronic databases, 58 orthopaedic systematic reviews were eligible for inclusion based on specific criteria. Thirty of these (52%) included nonrandomized studies, 15 of which found no randomized controlled trials. Systematic reviews were more likely to include randomized controlled trials if nondistinguishable operations were compared (if participants could be blinded). Only six of the systematic reviews that included nonrandomized studies (20%) assessed the quality of primary studies. Heterogeneity of studies was a major concern. In 21 of the systematic reviews that included nonrandomized studies (70%), data for groups treated similarly were pooled across studies, and outcomes for pooled groups were compared. The conclusions of systematic reviews that included nonrandomized studies are weakened by the limitations of nonrandomized study designs. The absence of established methods for including nonrandomized studies in systematic reviews, and consequently variability in the methods adopted, also limits the comparability of such reviews. Therefore the findings of systematic reviews that include nonrandomized studies should be interpreted with caution. PMID- 15552166 TI - Elbow soft tissue mass in a 9-year-old boy. PMID- 15552167 TI - Posttraumatic intracortical lytic lesion in a 46-year-old woman. PMID- 15552168 TI - A 32-year-old man with knee pain. PMID- 15552169 TI - Mortality after reamed intramedullary nailing of bilateral femur fractures. PMID- 15552170 TI - A healthy workplace promotes quality care. PMID- 15552171 TI - Continence clarification. PMID- 15552172 TI - Staff mix and public safety. PMID- 15552174 TI - Staff mix and public safety. PMID- 15552173 TI - Staff mix and public safety. PMID- 15552175 TI - Nurse practitioners. PMID- 15552176 TI - Setting the bar for rural nurse practice. PMID- 15552177 TI - Changing staff mix. PMID- 15552178 TI - Finding the right balance. PMID- 15552179 TI - Developing nursing's future leaders. PMID- 15552180 TI - Improving post-operative pain management by focusing on prevention. PMID- 15552181 TI - Quality practice environments are in the public's interest. PMID- 15552182 TI - For a limited time only. Left in the lurch, desperate hospitals are turning to interim healthcare managers to temporarily fill executive positions. AB - A convergence of factors in the healthcare industry has helped create a new career track-the interim CEO. It's a demanding but enjoyable path for some. "You've got to be willing to live the lifestyle," says Lowell Johnson, left, who's worked for 22 employers in the past 16 years. PMID- 15552183 TI - National ambition. Deal would put Coventry in all 50 states. PMID- 15552184 TI - Tenet selling three in Mass. PMID- 15552185 TI - Spreading the wealth. HHS technology grants target rural communities. PMID- 15552186 TI - Getting personal. Cerner offers $25 million to create health records. PMID- 15552187 TI - Coming up short. Shortages of space, beds, staff cause capacity crunch. PMID- 15552188 TI - When doses disappear. Flu shot shortage sheds light on drug supply chain. PMID- 15552189 TI - Drug pullout. Massive recall of Vioxx poses logistical problems. PMID- 15552190 TI - Out of sight. Pharmaceutical industry temporarily replaces hospitals as target. PMID- 15552191 TI - Designing with health in mind. Innovative design elements can make hospitals safer, more healing places. PMID- 15552192 TI - It's all by design. Award-winning projects make sure people come first. PMID- 15552193 TI - Straight talk. New approaches in healthcare. Outsourcing in the business office increases cash. AB - Want more cash? Rather than simply sending old patient balances to collection agencies in knee-jerk fashion, sophisticated business offices are scrutinizing their days in accounts receivable, determining what types of accounts and dollar balances to outsource for collection and when to outsource them. In this installment of Straight Talk, we look at the outsourcing strategies in the business offices of three healthcare systems: 266-bed Tuomey Healthcare System, Sumter, S.C.' 3-hospital, 1,062-bed Montefiore Medical Center, New York; and 14,200-bed, 30-hospital New York Presbyterian Health System, New York. Modern Healthcare and PricewaterhouseCoopers present Straight Talk. The session on outsourcing of business office functions was held on September 23, 2004 at Modern Healthcare's Chicago headquarters. Charles S. Lauer, publisher of Modern Healthcare, was the moderator. PMID- 15552194 TI - By the numbers. Top 10 healthcare provider consultants: ranked by 2002 provider revenue. PMID- 15552195 TI - Internal customer service: caring for each other at work. PMID- 15552196 TI - EMTALA: the ophthalmic nurse as qualified medical personnel (QMP). PMID- 15552197 TI - Topical NSAIDS to control pain in clear corneal cataract extraction. AB - This study was conducted to assess the additive efficacy of ophthalmic topical nonsteroidal anti-inflammatory drugs (NSAIDs) with topical anesthesia in the control of pain associated with clear corneal cataract extraction. The patients who received three days of preoperative topical NSAIDs had a statistically significant decrease in their level of discomfort. We have previously shown that three days of preoperative NSAIDs can reduce postoperative inflammation after cataract surgery. This study demonstrates that ophthalmic topical NSAIDs also decrease discomfort during surgery. PMID- 15552198 TI - Bacterial contamination rate of eyedrops: comparison of a hospital and a private outpatient center in Kermanshah, Iran. AB - Bacterial contamination rate of eyedrops: comparison of a hospital and private outpatient ophthalmology centers of Kermanshah, Iran. Eye drops are sterile solutions with different clinical effects. Like any therapeutic medication, they should be administered using aseptic technique and stored in a way that prevents bacterial contamination. In our hospital and local private clinical settings, some cases of bacterial keratitis are thought to be due to eyedrops, which are used on multiple patients and at some point become contaminated. This study was done to compare the bacterial contamination rate in an educational hospital with private ophthalmic centers and attempt to identify the route of contamination and consequently, a method to resolve or reduce the problem. PMID- 15552199 TI - Cyber sight--ORBIS telemedicine. PMID- 15552200 TI - Ophthalmic coherence tomography (OCT). PMID- 15552201 TI - Allergic to PVP? A discussion of alternatives. PMID- 15552202 TI - Flying engineer teaches thousands to see with one eye: the 30-year background of the book, A Singular View. PMID- 15552203 TI - Demonstrate your competency with the CRNO credential! PMID- 15552204 TI - Wilmer eye institute nurses create AMD support group initiative learn more at the ASORN annual meeting. PMID- 15552205 TI - Cultural pathways through the information age. PMID- 15552206 TI - Expressive communication and human development in the new broadband environment. PMID- 15552207 TI - How children and adults learn to use computers: a developmental approach. PMID- 15552208 TI - Emotional capital and information technologies in the changing rhetorics around children and childhoods. PMID- 15552209 TI - [Etiology of severe/profound, pre/perilingual bilateral hearing loss in Cantabria (Spain)]. AB - OBJECTIVE: To know the etiology of preiperlingual bilateral hearing loss in children. MATERIALS AND METHODS: All the patients diagnosed with bilateral severe/profound, pre or perilingual hearing loss at Sierrallana and Marques de Valdecilla Hospitals (Cantabria, Spain) during the last 20 years were included in this study. RESULTS: A hundred patients were diagnosed with bilateral severe/profound pre/perilingual hearing loss. The most frequent etiology was hereditary (49%), followed by severe perinatal hypoxia (11%), ototoxicity (5%), meningitis (3%), hyperbilirubinemia (3%) and rubella (2%). In 21% of cases was not known. CONCLUSIONS: The two most frequent etiologies found in severe/profound hearing loss in children in our area were hereditary and non infectious perinatal problems. Infectious disease were scarce. Will decrease when genetic test were used as clinical basis. PMID- 15552210 TI - [Blood viscosity modifications in patients with presbycusis]. AB - OBJECTIVE: [corrected] To evaluate the existence of blood viscosity alterations in presbyacusis as an eventual pathophysiological mechanism. MATERIAL AND METHODS: Nine 75 year-old subjects bearing of bilateral slowly progressive sensorineural deafness were considered to be studied on a yearly basis in a ten year folow-up period, evaluating the binaural hearing loss in speech tones (500, 1000, 2000 and 3000 Hz). Hemorheological profile on peripheral blood included total blood viscosity (BV) at different shear rates (SR), blood filterability (BF), erythrocyte deformability and aggregability (EA). RESULTS: There was a trend of all parameters towards hyperviscosity, with an increase at ten years of BV at high SR and EA at total stasis which were statistically significant. The level of deafness correlated significativelly to low values of BF and increases on BV at high SR. CONCLUSIONS: Presbyacustic patients do show hemorheological evidences that suggest a tendency towards hyperviscosity as a predisposing factor for hearing loss. PMID- 15552211 TI - [Prevalence of exostoses among surfers of the Basque Coast]. AB - Exostoses of the External Auditory Canal are benign bony tumours very common in individuals who frequently practice water activities. The objective [corrected] of this study is to determine the prevalence of exostoses in a population of surfers of the Basque Coast. We have studied the correlation between years surfing and the prevalence and severity of exostoses. We have done a cross sectional epidemiological study giving a questionnaire Twenty-five of the 41 surfers had surfer's ear (61%) and the prevalence was significantly higher in those who had surfed for more than 10 years (p < 0.05). Severity was also higher in the first group (p < 0.05). The relation between the hours dedicated to the sport showed similar results. only one of the 24 cases needed surgery (4%). PMID- 15552212 TI - [Endoscopic surgery of nasopharyngeal angiofibroma]. AB - Juvinile nasopharyngeal angiofibromas are highly vascular benign tumors but with an agressive behaviour with invasion of skull base and vital structures. Our study includes 11 male patients that underwent surgery for angiofibromas between 1992-2002 with an endoscopic surgical approach and embolization before surgery. The average age was 14.7 years old. Using Fisch staging, 8 patients were stage II. All cases achieved symptomatic remission, with no complications but the recurrence was 36.3%. The endoscopic excision is a safe and effective minimally invasive surgical treatment in patients in stage I and II. It should be considered as a first-choice option for these cases. PMID- 15552213 TI - [Laryngo-tracheal estenosis in adults. A restrospective study of 21 cases and literature review]. AB - We are reporting a retrospective study of 21 adults diagnosed of laryngo-tracheal stenosis and admitted to our Department between 1990 and 2001. In 9 cases (42.85%) it was due to prolonged orotracheal intubation; other 10 were secondary to tracheostomy (47.62%) and the rest, to coniotomy. We have studied and analysed these variables: Age, gender, personal history, level of stenosis, its grade according to Cotton's clasification, treatment and clinical evolution with an average follow-up of 12 months. Our main goal was to get people to know on experience with this pathology and to carry out a literature review in order to compare our data and results with other published series. PMID- 15552214 TI - [Quality of life in patients treated for early laryngeal cancer, in healthy people and in those with other associated pathologies]. AB - Nowadays there are different non aggressive treatment options for early laryngeal cancer. However, they are not free of undesirable effects. We have studied this patients' quality of life, by comparing it with that obtained from healthy population and other illness groups (post-AMI patients, Chronic Kidney patients). We have studied 62 T1-T2NoMo laryngeal carcinoma diagnosed at the "Virgen de la Victoria" clinical hospital from Malaga, Spain, between 1990 and 1998. We used the Quality of Life in Malaga (QLM), general QL questionnaire which was developed and validated at the Department of Preventive Medicine of the University of Malaga, Spain. The results were compared with those obtained from ramdom samples in healthy population from Malaga and from various pathology groups. The global QOL of our patients treated for an early glottic cancer (Index Quality of life 29.4%) was better than almost all the other ones. PMID- 15552215 TI - [Hodgkin and Non-Hodgkin lymphomas in otorhinolaryngology]. AB - Lymphomas of the head and neck arise in lymph nodes and extranodal areas as Waldeyer ring, nasal cavity, thyroid gland and salivary glands. Though anatomically in close proximity, lymphomas presenting at the ENT area have different clinical characteristics if they are Hodgkin or Non-Hodgkin. We included 31 patients with the diagnosis of lymphoma and diagnosed in our Department between 1999 and 2002. We studied all the different variables that differentiate Hodgkin and Non-Hodgkin's lymphoma. Both types of lymphoma usually present as a cervical mass. Non-Hodgkin lymphoma have extranodal involvement more frequently and also a more advance disease at diagnosis. It is important to include the lymphoma in the differential diagnosis of any cervical mass or ENT lesion. PMID- 15552216 TI - [Malignant laryngeal fibrous histiocytoma (MLFH). Report of two unusual cases]. AB - Laryngeal presentation of Malignat Fibrous Histiocytoma (MFH) is uncommon. It is more prevalent in elderly males. The most frequent form of laryngeal presentation is as subepithelial nodules, and its clinical behaviour is variable and unpredictable. Microscopically, it is a tumour with two well differentiated components: histiocitic and fibroblastic, with several different structural patterns that can make histologic diagnosis a difficult one. We present two cases of MFH of the larynx, and a review of the literature. PMID- 15552217 TI - [Malignant neck paraganglioma with a rare localization]. AB - We report a clinical case of a man with a cervical paraganglioma, originated from ectopic paraganglionic tissue, with metastasis to cervical lymph nodes. We carry out a reviw of the symptoms, diagnosis and treatment of this type of neoplasm. PMID- 15552218 TI - Non-HDL cholesterol predicts coronary heart disease in primary prevention: findings from an Italian 40-69 year-old cohort in general practice. AB - OBJECTS: Non-HDL cholesterol is now recommended as an index of risk associated with combined dyslipidemia, and it has also been found useful in predicting coronary heart disease (CHD) risk in patients with diabetes. We studied the association between known CHD risk factors, enclosed non-HDL cholesterol, and a "high CHD risk condition", i.e. a "5-years CHD risk >15%" in general practice. METHODS: We studied 4,085 40-69 year-old diabetic (no. 489) and non-diabetic (no. 3,596) individuals from an opportunistic cohort. Cross-sectional descriptive statistics, and age- and gender-adjusted multiple logistic exponential betas have been calculated. RESULTS: About 12% of the participants had diabetes. Age- and gender-adjusted comparison showed that all the study variables were significantly worse in diabetic vs. non-diabetic individuals (except cigarette smoking, total blood cholesterol and the ratio of total to HDL cholesterol). They had a mean "5 year CHD-risk" significantly higher than non-diabetic individuals (18.8+/-11.9% vs 7.5+/-6.9%, P<0.01), and a four-fold prevalence of "5-years CHD risk >15%" (55.4% vs 11.1%, P<0.01). As to diabetic individuals, the study variables associated to a "high CHD risk condition" were cigarette smoking, systolic blood pressure, and non-HDL blood cholesterol levels. As to non-diabetic individuals cigarette smoking, systolic blood pressure, and HDL (inversely) and non-HDL blood cholesterol levels were associated to a "high CHD risk condition". CONCLUSIONS: Non-HDL cholesterol--and cigarette smoking and systolic blood pressure--strongly predicted a "high CHD risk condition" both in diabetic and non-diabetic individuals. PMID- 15552219 TI - [The diastolic function: an echocardiographic study and its clinical meaning]. AB - Many patients with manifestations and/or symptoms of heart failure show a normal or only slight depressed systolic function of the left ventricle. The origin of these symptoms is attributed to anomalies of the diastolic function and the echocardiography has became an important technique for a non-invasive investigation of such anomalies. The valuation of the diastolic function, mainly in patients with a left ventricle dysfunction, is usually performed by means of the analysis of the transmitral flow with pulsed Doppler, integrating as necessary the basal information with actions of preload variations. However, in many cases it is necessary to use other procedures of study to better characterise the diastolic function in each patient by means of the analysis of venous pulmonary flow, the valuation of propagation velocity of mitral flow with Color M-Mode, or the valuation of diastolic velocities of myocardium with Doppler tissue imaging. The integrated analysis of these various methods of exam, moreover, allows to enhance the diagnostic accuracy in the evaluation of the filling pressure, which is a fundamental parameter to stratify the risk of short/medium term events, being equal the left ventricle dysfunction. PMID- 15552220 TI - [Heart failure: preventing rehospitalizations by disease management programs]. AB - Chronic heart failure is a growing public health problem for prevalence, morbidity and costs. The major proportion of costs is attributable to rehospitalizations and many of these readmissions may be preventable. Since 1990, some investigators have tested a variety of disease management programs designed to improve quality of life, functional status and decrease rehospitalizations rates. We identified these studies by a computerized search of the MEDLINE database. The programs described reflected a wide variety of methods and we categorized these programs recognizing the prevalent disease management approach. We reported the results of these trials about rehospitalizations and analysed a number of limitations that must be considered when determining their adoption into clinical practice. PMID- 15552221 TI - [Postoperative atrial fibrillation: etiopathogenesis, prevention and therapy]. AB - Atrial fibrillation is the most frequent complication after cardiac surgery. Its onset leads to a threefold higher risk for stroke compared with patients in sinus rhythm and other adverse events such as thromboembolic events and heart failure. The direct consequence is an increased length of hospital stays with obvious economic implications. These reasons have led the attention of many investigators to point out the rule of possible predisposing factors and underlining mechanisms in order to establish an effective preventive treatment. The present paper is aimed to review the state-of-the-art knowledge about post-operative atrial fibrillation and its complex etiopathogenesis which is in turn responsible for the lack of consensus regarding routine prophylaxis. PMID- 15552222 TI - [Nursing role in cardiac prevention]. AB - The lack of a multidisciplinary approach is certainly among the causes of the ineffectiveness of intervention in the field of cardiovascular secondary prevention. By multidisciplinary approach is meant involving cardiologists, nurses, rehabilitation therapists, dieticians as well as psychologists in the framing of interventions tailor made to patients needs. In particular, people working in the nursing area can play a very important role which can be summed up into three different levels: a technical level, aiming at the cooperation with cardiologists to carry out diagnostic examinations and give a portrait of patients in terms of risks; a second level consists in giving information, and helping to face the disease, as well as stepping in during its evolution, almost a health counsellor for the patients; finally the nurse can act as a psychological support both to the patient and his/her family during acute illnesses and reassure him/her that he/she is being treated properly and that successively will resume a good quality of life. Hospital represent an ideal place for secondary prevention, at least in the first phases of the intervention. The results collected during hospitalization would be rapidly lost if they were not followed and sustained in the medium-long term by structured follow-up programmes. The development of ambulatories might represent a link between hospitals and the territory, i.e. the specialist and one's personal physician. The staff of ambulatories should comprise a cardiologist and a trained professional nurse, this being specialized, specifically, in cardiology and cardiovascular prevention. Staff of the type described could work independently, co-ordinating ambulatories on the territory within the framework of standardized recognized protocols and relating information concerning patients, general practitioners and other surgeries. In this way, an essential link of the continuity in medical care would be guaranteed. PMID- 15552223 TI - [Cardiovascular prevention and coronary heart disease mortality decrease]. PMID- 15552224 TI - [Gender medicine]. PMID- 15552225 TI - [Gender-specific medicine]. PMID- 15552226 TI - [The category gender and meeting standards for medical education as a result]. AB - The Medical School Hanover, Institute of History, Ethics and Philosophy in Medicine, in co-operation with the University of Bristol, School for Policy Studies, carried out a project to examine the meaning of gender in medical education and to detect methods to incorporate the gender-perspective from public health research and gender studies as an interdisciplinary programme into medical education. The duration of the project was limited to two years, and funded by the EU from 2001 to 2003. The study clearly shows that the category gender is being very slowly introduced into medical education at medical faculties and medical universities in Europe. Questions are primary focussed on biological differences between the sexes of patients, whereas gender studies touch gender relationships in occupations, interactions and communication, different styles of leadership etc. The adaptation of this approach may lead to a new understanding of the doctor-patient-interaction as well as of the relationship between teachers and students. PMID- 15552227 TI - Domestic violence--a comparison of activities in the health care sector in Brazil and Germany. AB - During the last decades an, international discussion has started on how to reduce the gender gap in women's and men's health. Feminist activists and public health scientists in Canada, Latin America (foremost Brazil), the States and South Africa issued a declaration of intention to focus on health impacts caused by gender-related domestic violence. In Europe, and particularly in Germany, the topic as an issue of medical care has lately been given increased attention. In this article, we present activities undertaken to overcome the obstacles in integrating the topic of domestic violence as a medical issue. The successful Brazilian strategy served as a model, in particular for implementing a hot-line for victims of domestic violence within the day-to-day clinical setting. The General Medical Council (Arztekammer), the Women's Medical Association of Lower Saxony, and the Hanover Medical School founded a working committee "domestic violence" in co-operation with the Ministry of Women, Work and Social Affairs. Between 2002 and 2003, this task force worked out several guidelines and hand outs for physicians to promote gender-sensitive treatment of abused women, gaining increasing acceptance by the German medical care providers in medical education and in-training services for physicians. PMID- 15552228 TI - Osteoporosis: gender-specific aspects. AB - Traditionally, osteoporosis has been regarded as a disease of postmenopausal women; nevertheless, although osteoporosis is more common in women than in men, osteoporosis in men is a frequent and a severe condition. Osteoporosis generally can be characterized as either primary or secondary; in men secondary osteoporosis is more frequent than in women. The role of estrogen deficiency in the pathogenesis of postmenopausal osteoporosis is clearly established; remarkably, evidence accumulated over the past years strongly suggests that estrogen also plays the dominant role in regulating the male skeleton. Several independent studies observed decreased serum estradiol levels in men with idiopathic osteoporosis. In contrast to postmenopausal osteoporosis, there are only few studies on the treatment of osteoporosis in men; two different compounds (alendronate and teriparatide) appear to be equally effective in men and women. PMID- 15552229 TI - [The role of gender medicine in rehabilitation]. AB - This article provides a coherent view of gender differences in rehabilitation medicine from the biological, individual and social perspective. It reports the role of gender in pain, in rehabilitation of the musculosceletal system and in rehabilitation of patients with prostate cancer. PMID- 15552230 TI - Do we diagnose and treat coronary heart disease differently in men and women? AB - There are gender-specific differences in the significance of cardiovascular risk factors, as well as in the symptoms and in the diagnostic approach of cardiovascular events. From the point of view of everyday clinical practice, the differential approach toward both genders is based on gender-specific risk assessment. A global risk assessment should be carried out in males > 40 years of age and in females > 50 years or those who are postmenopausal. Diabetes and hypertriglyceridemia require aggressive therapy particularly in women. Depending on level of risk appropriate therapy should be instituted: life style counseling (smoking!), therapy of dyslipidemias, antihypertensive therapy and diabetes control. Symptoms of coronary attack are experienced by men more often "classically", whilst women commonly present with unspecific symptoms, which may delay proper medical care. Appropriate patient education is needed particularly in younger women to avoid unnecessary delay of intervention in acute coronary syndromes. Regarding diagnostics, there are gender differences in the specificity and sensitivity of some noninvasive diagnostic tests, which should be taken into account. PMID- 15552231 TI - [Gender aspects of socioeconomic and psychosocial risk factors of cardiovascular diseases]. AB - Socioeconomic and psychosocial factors exert influence on health as well as the development, the progression and the prevention of diseases. Social factors regarding cardiovascular diseases have been widely researched. Whilst characteristics of classic type A behaviour increase cardiovascular risk among men, characteristics of type B behaviour represent a protective value, especially among women. Depression--a disease that is particularly prevalent among women and is associated with socioeconomic factors--negatively influences the development of cardiovascular diseases, triggers cardiovascular events and influences rehabilitation. Lifestyle factors, which are positively or negatively correlated with cardiovascular disease, show a gender-specific prevalence and are related to psychosocial factors. More women than men report healthy nutrition, whereas more men report partaking in physical exercise. Obesity is--depending on the occupational group and the social level--more prevalent among women compared to men. PMID- 15552232 TI - [The importance of the parameter sex & gender in anesthesia and intensive care medicine dealing with patients under alcohol influence]. AB - Alcoholised females and males belong to a specific group of risk patients in anesthesia and intensive care medicine. Knowledge about effects touching the interaction of chronic alcohol consumption and acute alcohol abuse with other drugs is on a very low level. The assessment of risk depends on several factors. In addition to the judgement of grades of seriousness of an alcohol intoxication the parameter of sex and gender can be seen as much a prominent parameter as age, morbidity, and co-morbidity. This thesis is confirmed by some significant sex based biological differences related to biological effects of alcohol consumption and differences in pharmacodynamics and pharmacokinetics of anesthetics. The high rate of injuries and accidents under the influence of alcohol, strongly recommends the detection of a sex- and gender-sensitive approach by anesthesia and intensive care medicine to patients with chronic or acute alcohol abuse. PMID- 15552233 TI - Looking toward the epidemic of obesity. PMID- 15552234 TI - Healthy children. PMID- 15552235 TI - Using BRFSS data to estimate prevalence of obesity and physical activities in Oklahoma. AB - Obesity has risen at an epidemic rate during the past two decades in the United States. OBJECTIVE: this report describes the obesity prevalence and its relations to physical activities in Oklahoma. METHOD: Used data from Behavioral Risk Factor Surveillance System 2001-2003 for analysis. RESULT: In Oklahoma, the prevalence rates of obesity were increasing in the recent decades and reached 23.3%. People with lower income, fewer years in school, being male, and certain minority groups tend to have higher obesity prevalence rate than others. People with obesity tend to report having chronic diseases. Although self-reported moderate physical activity in Oklahoma has already met the Healthy People 2010 Objective, percentage of no leisure-time physical activity was still high. CONCLUSION: Relationship between prevalence of obesity and physical activity is difficult to interpret. Other risk factors of obesity should be considered in further study. PMID- 15552236 TI - Low-carbohydrate diets: what should we recommend? PMID- 15552237 TI - Childhood obesity: assessing the cost. AB - This paper addresses the cost of childhood and adolescent obesity on the United States healthcare system. The specific focus is the prevention and treatment of obesity in children. The cost of prevention and treatment programs are considered as well as other costs due to further complications caused by obesity in children and adolescents. Examples of health expenditures in Canada are used to exemplify the prevalence of obesity and its effect on healthcare expenditures. PMID- 15552238 TI - Overview and management of obesity in adults. AB - Teaching Points. The prevalence of overweight or overtly obese adults in the United States is growing and associated with significant morbidity and mortality. Office evaluation of the overweight or obese adult includes a thorough history and physical examination complemented by several laboratory tests that evaluate for co-morbid conditions. Treatment of the overweight or obese adult requires a step-wise approach beginning with lifestyle modification followed by pharmacologic or surgical interventions in selected patients. Treatment decisions should take into consideration body mass index, waist circumference and the presence of comorbidities or obesity-associated risk factors. PMID- 15552239 TI - Childhood obesity: epidemiology, comorbid conditions, psychological ramifications, and clinical recommendations. AB - The prevalence of childhood obesity in the United States has increased dramatically in recent years, doubling in the last 20 years, and is expected to result in a major public health crisis. This epidemic has left no race, socioeconomic status, or age unaffected. Children in Oklahoma are no exception; according to the Centers for Disease Control and Prevention, Morbidity and Mortality Weekly Report, 25% of Oklahoma's adolescents are at risk for overweight or are overweight. In order to address this problem, efforts are underway nationally to institute preventive measures for childhood obesity and its comorbid conditions. PMID- 15552240 TI - Obesity in the elderly: survival of the fit or fat. AB - Obesity is an important health problem for the growing elderly segment of the population. Age-related changes in body composition should be taken into account when considering morbidity. Today, sarcopenic obesity, which is defined as excess fat with loss of lean body mass, is a highly prevalent problem. Obesity in the elderly is related to morbidity; e.g., sleep apnea, cancer, osteoarthritis, diabetes and hypertension. The advantages and disadvantages of using BMI, waist circumference, waist: hip ratio, and body weight to measure age-related changes in obesity are discussed. In addition, the merits of treatment options for obesity; e.g., behavioral modifications, diet, and exercise--are described. One important conclusion derived from a review of these treatments is that age itself is not a contraindication for pharmacotherapy or even surgery for morbid obesity. PMID- 15552241 TI - Association of obesity and smoking with chronic diseases among Oklahoma adults. AB - Obesity and smoking are the two modifiable risk factors that contribute the most for many chronic diseases. Obesity has nearly doubled in Oklahoma and in the US since 1990 while Oklahoma adult smoking rates have remained above 25%. The purpose of this study was to utilize the Behavioral Risk Factor Surveillance System (BRFSS) to describe the associations between overweight, obesity, and cigarette smoking with selected chronic diseases among Oklahoma adults. The BRFSS is an ongoing, state-based, random-digit dialing telephone survey of the non institutionalized adult population ages 18 years and older designed to collect various kinds of health-related information. Descriptive statistics were obtained for high blood pressure, high cholesterol, diabetes, asthma, and arthritis by BMI status (i.e. not overweight, overweight, obese) and smoking status (i.e., current, former, never). Logistic regression was performed to test for associations between BMI status or smoking status and these diseases. Self reported diabetes, arthritis, and lifetime high blood pressure and high cholesterol rates were higher among those who were overweight and highest among those who were obese, while asthma rates were higher among obese adults than those who were not obese. Asthma rates were higher for those who currently smoke than former smokers and never smokers. Former smokers had significantly higher rates of high blood pressure, high cholesterol, diabetes, and arthritis compared to never smokers or current smokers. PMID- 15552242 TI - High-energy, high-return practice. PMID- 15552243 TI - Determining a proper sample size. PMID- 15552245 TI - Putting the family in family medicine. PMID- 15552244 TI - Evaluating clinical literature. PMID- 15552247 TI - Investing in IT could get easier. PMID- 15552246 TI - Understanding when to use modifier -25. PMID- 15552248 TI - Can you negotiate better reimbursement? PMID- 15552249 TI - An introduction to tablet PCs. PMID- 15552250 TI - Asthma days: an approach to planned asthma care. PMID- 15552251 TI - A tool for evaluating patients with cold symptoms. PMID- 15552252 TI - The big deal about small talk. PMID- 15552253 TI - Senate Appropriations Committee would halt funds - to implement "75 percent rule" for rehab facilities. PMID- 15552254 TI - Kahn: physician-owned limited service facilities "getting in under the wire". PMID- 15552255 TI - Premiums grow 11.2 percent in 2004; employers project 0 9.6 percent hike for per employee costs. PMID- 15552256 TI - Health care at a cross-road. PMID- 15552257 TI - Making health care more accessible and affordable. PMID- 15552258 TI - Conflict of interest hurts patients. PMID- 15552259 TI - Laser,"closure", stents and other new technology in the treatment of venous disease. AB - New technology is transforming our approach to venous disease. Once considered taboo for fear of causing deep venous thrombosis, surgical or endovenous intervention in the deep venous system is now known to be safe. Newer techniques allow minimally invasive procedures, many carried out on an outpatient basis. Traditional conservative regimens to treat acute ilio-femoral DVT are yielding to more aggressive interventional approaches to minimize the high incidence of postthrombotic syndrome with the former. Stent technology can safely and effectively relieve many of the disabling symptoms of chronic venous obstruction. Stent patency is high and morbidity low. Knowledge of venous pathophysiology has also advanced, though much remains to be learned. The beneficiaries are the large patient population with DVT and chronic venous insufficiency, which is estimated to be at least three times as large as patients with arterial disease. Unlike arterial disease, venous pathology afflicts the younger working age population in large numbers at enormous direct and social costs. In the older patient, deep venous disease is common, but seldom suspected. The symptoms are often ascribed to systemic causes. A thorough venous investigation is the key to proper diagnosis and treatment. Often, a minimally invasive procedure such as EVLT or stent insertion can offer surprising symptom relief with significant improvement in the quality of life during the twilight years. PMID- 15552260 TI - The uninsured. PMID- 15552261 TI - "What I did this summer" revisited. PMID- 15552262 TI - Some pediatric excerpts and observations in 1910. PMID- 15552263 TI - What physicians need to learn from veterinarians. PMID- 15552264 TI - The significance of serum gamma-glutamyltransferase in cardiovascular diseases. AB - Since early after the introduction of serum gamma-glutamyltransferase (GGT) in clinical practice as a reliable and widely employed laboratory test, epidemiological and prospective studies have repeatedly shown that this activity possesses a prognostic value for morbidity and mortality. The association is independent of possibly concomitant conditions of liver disease, and notably, a significant independent correlation of serum GGT exists with the occurrence of cardiovascular diseases (myocardial infarction, stroke). Experimental work has documented that active GGT is present in atherosclerotic plaques of coronary as well as in cerebral arteries. These findings, and the recently recognized functions of GGT in the generation of reactive oxygen species, indicate that serum GGT represents a true marker of cardiovascular diseases and underlying atherosclerosis. Further insights into potential therapeutic interest will probably be derived from studies investigating the origin of GGT activity in plaque tissue. PMID- 15552265 TI - Anti-tissue transglutaminase antibodies in inflammatory bowel disease: new evidence. AB - Anti-tissue transglutaminase, previously held to be identical to anti-endomysial antibodies in celiac sprue, has been reported in inflammatory bowel disease patients. To investigate these data further, we evaluated serum and intestinal anti-tissue transglutaminase in inflammatory bowel disease patients, with respect to the Crohn's disease activity index and the integrated disease activity index. Study population comprised: 49 patients with Crohn's disease and 29 patients with ulcerative colitis; 45 patients with celiac sprue and 85 autoimmune patients as disease controls; and 58 volunteers as healthy controls. Immunoglobulin A (IgA) anti-recombinant human tissue transglutaminase and anti-endomysial antibody detection in sera and fecal supernatants were performed. Adsorption of positive sera with recombinant human tissue transglutaminase were also performed. Marked increased anti-tissue transglutaminase concentrations were found in celiac sprue, while low-positive values were also found in Crohn's disease and ulcerative colitis. Anti-endomysial antibodies were detectable only in celiac sprue. Antigen adsorption resulted in a significant reduction of the anti-tissue transglutaminase either in celiac sprue or inflammatory bowel disease sera. A significant correlation between anti-tissue transglutaminase and Crohn's disease activity index or integrated disease activity index scores was found. Anti-tissue transglutaminase was also detectable in fecal supernatants from inflammatory bowel disease patients. Data highlight that both circulating and intestinal anti tissue transglutaminases are detectable in inflammatory bowel disease, and that they are related to disease activity. These features underline that, in addition to anti-tissue transglutaminase, an anti-endomysial antibody test is necessary in the diagnostic work-up of celiac sprue, especially in patients with known inflammatory bowel disease. PMID- 15552266 TI - Expression level of MDR1 message in peripheral blood leukocytes from healthy adults: a competitive nucleic acid sequence-based amplification assay for its determination. AB - Accurate quantification of multidrug resistance-1 gene (MDR1) expression in target cells would be of important therapeutic value in predicting cellular response to anticancer drugs. Because certain normal cells in peripheral blood physiologically express MDR1, increasing the sensitivity of the detection methods might result in confounding low-degree expression in tumor cells with physiologic expression in normal cells. The purpose of this study was to determine MDR1 mRNA expression levels in peripheral blood leukocytes obtained from healthy adult volunteers using a competitive nucleic acid sequence-based amplification (NASBA) assay. We determined the reference intervals of MDR1 mRNA expression in peripheral blood obtained from 98 healthy adults by measuring its expression with the quantitative NASBA assay between 5.50 x 10(4) copies/microg RNA and 6.76 x 10(5) copies/microg RNA. The new reference intervals were evaluated using a number of sensitive or resistant cell lines as control; positive or negative MDR1 expression was clearly demonstrated. We also reevaluated MDR1 expression levels in leukemia cells obtained from patient peripheral blood; 18 of 31 samples (58%) exceeded the newly established upper reference limit. The cutoff value established could be used to distinguish significant MDR1 expression in tumor cells from physiologic expression in certain normal cells coexistent in peripheral blood. PMID- 15552267 TI - Characteristics of prolinase against various iminodipeptides in erythrocyte lysates from a normal human and a patient with prolidase deficiency. AB - The effect of various amino acids and MnCl2 on prolinase activity in erythrocyte lysates from a healthy individual and a patient with prolidase deficiency was investigated. A concentration of 0.1 mM MnCl2 increased prolinase activity in normal erythrocytes against pro-gly, pro-glu, pro-leu, pro-ser and pro-phe, but inhibited that against pro-ala, pro-val, pro-met and pro-asp. However, prolinase activity against these iminodipeptides was enhanced by pre-incubation with glycine, independent of MnCl2. The same studies on erythrocytes from a prolidase deficient patient showed almost the same results as the normal control, except that prolinase activity against pro-gly and pro-ser was slightly inhibited by adding 0.1 mM MnCl2. Some amino acids, glutamic acid and glutamine, slightly enhanced prolinase activity against pro-gly in erythrocytes from both the normal control and the prolidase-deficient patient, but N-acetyl-L-glutamic acid, gamma aminobutyric acid (GABA) and beta-alanine showed no effect. Branched amino acids, L-valine, L-leucine and L-isoleucine strongly inhibited the prolinase activity against pro-gly. However, conversely, their isomers, D-valine, D-leucine and D isoleucine, enhanced it. The kinetics of prolinase activity in the erythrocytes from both the normal individual and the prolidasedeficient patient were also studied. Their Km values were changed by adding glycine or 0.1 mM MnCl2, but Vmax values were almost the same. PMID- 15552268 TI - Alteration of homocysteine catabolism in pre-eclampsia, HELLP syndrome and placental insufficiency. AB - Hyperhomocysteinemia is a risk factor in obstetrical complications such as pre eclampsia, 'hemolysis, elevated liver enzymes, low platelet' (HELLP)-syndrome and placental insufficiency. The aim of our study was to investigate the alterations of homocysteine catabolism in these patients in relation to serum B-vitamins and renal function. Maternal fasting serum from pre-eclampsia (n=24), HELLP (n=20) and placental insufficiency (n=25) patients at the time of diagnosis and pregnant controls (n=34) was analyzed for homocysteine and its metabolites cystathionine and methylmalonic acid, the vitamins B6, B12 and folate, renal and additional parameters. Cystathionine, a parameter of homocysteine catabolism, was significantly increased in pre-eclampsia and HELLP compared with controls and placental insufficiency patients (mean concentrations: 343, 324, 248, 227 nmol/l; p=0.001). Homocysteine, folic acid, vitamin B6 and methylmalonic acid, however, did not differ significantly between groups. The main determinants of cystathionine are cystatin C and vitamin B6, whereas the main determinants of homocysteine are folate and uric acid. The strongest dependency of cystathionine on vitamin B6 was observed in pre-eclampsia and HELLP patients. The results suggest that the vitamin B6-dependent trans-sulfuration pathway is activated in pre-eclampsia and HELLP syndrome, probably by oxidative stress. Therefore, the demand for vitamin B6 is increased in these patients. Furthermore, renal dysfunction and low vitamin B6 levels contribute to the increase of cystathionine in pre-eclampsia and HELLP patients. PMID- 15552269 TI - Immune system-mediated endothelial damage is associated with NO and antioxidant system disorders. AB - Two distinct systems of different origin are involved in the pathogenesis of both infectious and immunological vasculitis syndrome: nitric oxide (NO) from endothelial cells and granulocyte NADPH oxidase. In this study, in 31 children with immune system dysfunction, NO, NO synthase (NOS) and antioxidant enzyme activities [catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx)], as well as immunological parameters, were investigated. On the basis of the clinical findings, all children were divided into three groups: group I, 8 children clinically showing macular skin manifestations; group II, 11 children with maculo-papulous changes; and group III, 12 children with clinical findings of papulous changes. Plasma NO values in groups II and III were significantly elevated (79.14+/-30.13 and 65.32+/-6.70 micromol/l), compared to the control group (41.24+/-3.65 micromol/l), while group I showed statistically lower values (32.38+/-3.37 micromol/l). In children with the highest level of NO (group II) NOS activity was two-fold higher (1.77+/-0.59 nmol/ml/min; p<0.01) than in controls (0.98+/-0.23 nmol/ml/min). Catalase activity showed a significant increase and SOD activity a significant decrease in all experimental groups, while GPx was not significantly changed. The results show that immune system dysfunction manifested as vasculitis is associated with significant disturbances in the NO system and free radicals scavengers. PMID- 15552270 TI - Bone marrow microvascular density and angiogenic growth factors in multiple myeloma. AB - There is evidence that angiogenesis plays an important role in the progression of multiple myeloma (MM). Hepatocyte growth factor (HGF) and tumor necrosis factor alpha (TNF-alpha) are cytokines that potently stimulate angiogenesis. We evaluated the microvascular density (MVD) of bone marrow biopsies (after immunostaining with anti-CD34 antibodies) and serum levels of HGF and TNF-alpha in 43 patients with newly diagnosed MM. Twenty-four of these patients reached a plateau phase after treatment and were reevaluated for MVD, HGF and TNF-alpha. MVD values and serum levels of HGF and TNF-alpha were elevated in newly diagnosed MM patients in comparison with healthy controls. Pre-treatment MVD, HGF and TNF alpha increased with advancing stage of MM disease. In patients reaching the plateau phase, a significant reduction in MVD, HGF and TNF-alpha levels occurred. A positive correlation was noted between pre-treatment MVD and serum levels of TNF-alpha and lactic dehydrogenase but not with HGF. However, HGF strongly correlated with beta2-microglobulin (beta2M), TNF-alpha and lactate dehydrogenase (LDH). We conclude that angiogenesis in MM, as expressed by the bone marrow MVD and the serum levels of angiogenic molecules such as HGF and TNF-alpha, increases with advancing clinical stage and decreases after effective chemotherapy. PMID- 15552271 TI - The insulin-like growth factor system in the circulation of patients with viral infections. AB - The insulin-like growth factor (IGF) system was examined in the circulation of patients with viral infections (herpes simplex virus, HSV; cytomegalovirus, CMV; rotavirus, RV and adenovirus, AV). The serum concentrations of IGF-I, IGF-II and cortisol were measured by radioimmunoassay, while IGF-binding proteins (IGFBPs) were characterised by ligand-affinity blotting. Although both IGF-I and IGF-II concentrations were significantly lower in patients with viral infections (p<0.05) than in healthy persons, the IGF-II/IGF-I ratio was increased (p<0.05). No correlation between the concentration of IGF-I and IGF-II and the intensity of the antibody response to infection was observed. Ligand-affinity blotting demonstrated decreased amounts of IGFBP-3 (patients with HSV, CMV, AV and some patients with RV), increased IGFBP-2 (some patients with HSV and RV) and IGFBP-1 (patients with RV). Serum cortisol was significantly elevated (p<0.05) in patients infected with HSV, CMV and RV. The alterations observed can be interpreted as induction of the hypothalamic-pituitary-adrenal axis and suppression of the growth hormone (GH)/IGF axis under the influence of viral infection. PMID- 15552272 TI - Time course of systemic markers of inflammation in patients presenting with acute coronary syndromes. AB - Inflammation within coronary plaques may cause an acute coronary syndrome by promoting rupture and erosion. It was the aim of this study to examine whether markers of inflammation derive from a cardiac or extracardiac source and how their levels develop over time. Blood samples were taken from patients with acute coronary syndromes (ACS) with proven atherosclerotic lesion(s) of the left coronary artery (n=13) and from control patients without coronary artery disease (n=13). Blood was taken from the femoral vein and the coronary sinus vein before and after coronary angioplasty (day 0) and on days 1 and 120. Levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-1-receptor antagonist (IL-1 ra) and soluble CD40 ligand (sCD40L) were higher in ACS patients as compared to controls and remained elevated up to day 120. In the long-term time course these markers of inflammation and plaque remodeling slightly decreased in ACS patients. There were no statistically significant differences detectable in the levels of TNF-alpha, IL-6, IL-1 beta, IL-10, IL-1 ra, sCD40L and monocyte chemoattractant protein-1 (MCP-1) in the blood of ACS patients taken from a cardiac source as compared to an extracardiac source (coronary sinus vs. femoral vein). This study demonstrates the importance of a systemic inflammatory condition in patients with ACS, in whom markers of inflammation are increased as compared to controls. During long-term follow-up the pro-inflammatory activity remains elevated in ACS patients, supporting the concept of a systemic rather than a local vascular inflammation contributing to the development of atherosclerosis. PMID- 15552273 TI - Age- and sex-related reference values for serum insulin concentration and its biological determinants in a French healthy population. The STANISLAS cohort. AB - Insulin is involved in coronary heart disease through diabetes and metabolic syndrome. A great deal is known about insulin and its correlates, as well as factors related to changes in insulin. However, few studies consider the broad variety of correlates simultaneously. Therefore, the aims of the present study were to characterize the main factors of biological variation affecting serum insulin concentration and to establish reference limits of insulinemia in a presumably healthy French population. Insulin was measured using a microparticular enzymatic immunoassay. A total of 646 subjects aged 11-58 years from the STANISLAS cohort and divided into four groups of 162 males, 157 females, 163 boys and 164 girls, were included in the statistical analyses. In the whole population, serum insulin concentration varied from 0.80 to 54.60 microU/ml. Significant factors affecting insulin were age, gender, body mass index and glucose, in addition to alanine aminotransferase and high-density lipoprotein cholesterol in men, triglycerides and oral contraceptive use in women, and alkaline phosphatase in girls. In summary, we presented biological correlates of insulin in both healthy French male and female adults and children/adolescents and determined reference limits for insulin for each group. These results will contribute to a better interpretation of insulin data in further studies and laboratory investigations. PMID- 15552274 TI - Potential pitfalls of comparative measurements of reticulocytes with flow cytometry and microscopy in prematures and infants. AB - Reticulocyte counting by flow cytometry (Bayer H*3, ADVIA 120) in blood of prematures, infants and children > 1 year of age was compared with microscopic counting under research conditions (9000 counted red blood cells per slide). While in children >1 year a good concordance of both methods was observed, 2.3 2.4-fold higher values were obtained in neonates by microscopy (Brilliant Cresyl Blue stain, 0.5%). However, another laboratory found good agreement between H*3 counting and microscopy in samples also obtained from neonates using the same methods. Despite very similar results for all age groups in comparative flow cytometry measurements in both laboratories, counting of smears from neonates differed, showing an approximately 2.3-fold larger amount of reticulocytes in our laboratory. The reason for these observations was a greater enlargement (1250 fold) used routinely in our laboratory compared with 800-fold in the other one. Thus very mature reticulocytes frequently found in neonates could only be detected using a 1250-fold enlargement. Similarly, the low concentration of the colouring matter used in the H*3 (0.0005% oxazin or 0.001% ADVIA 120) is obviously not sufficient for detection of mature reticulocytes. Therefore, it is important to consider this phenomenon and to standardise microscopic enlargement, especially for comparisons in multicentre studies. PMID- 15552275 TI - Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay. AB - Current clinical practice considers antinuclear antibody (ANA) testing as a screening test; this has a major impact on laboratory work with a growing volume of analyses that need to be performed rapidly, to maintain good specificity and sensitivity. Ongoing discussions have been raised in order to identify the best technology to use in ANA screening, taking into account both clinical and economical implications. The aim of our study was to compare three different enzyme immunoassays (EIA) with immunofluorescence (IF) assay in order to identify which test is better for use as a screening test. The study was performed on 473 sera and the three different EIA tests were based on nuclear homogenates from HeLa cells, purified antigens from HEp-2 cells and recombinant antigens, respectively. The concordance between EIA-ANA and IF-ANA techniques, determined by the K statistic, was acceptable, but not complete, and discrepancies between both EIA-positive/IF-negative samples and IF-positive/EIA-negative were found. Both methods show interesting diagnostic abilities, however, the IF-ANA assay seems to be the first choice test in a well-standardized immunofluorescence laboratory with experienced microscopists, whereas the EIA test might be useful especially in large-scale ANA screening. PMID- 15552276 TI - Analytical and diagnostic accuracy of the EliA automated enzyme fluoroimmunoassay for antineutrophil cytoplasmic autoantibody detection. AB - Anti-proteinase 3 antineutrophil cytoplasmic antibodies (PR3-ANCA) and anti myeloperoxidase antibodies (MPO-ANCA) are considered important serological markers for several forms of idiopathic systemic vasculitis. The aim of the study was to verify the analytical and clinical performance of a new automated enzyme fluoroimmunoassay, the EliA system, for PR3-ANCA and MPO-ANCA detection. For this purpose the sera of 52 consecutive well-defined patients with a clinical diagnosis of Wegener's granulomatosis (WG) (n=29) or microscopic polyangiitis (MPA) (n=23), and 70 controls suffering from connective tissue disease (25 systemic lupus erythematosus, 25 Sjogren's syndrome and 20 rheumatoid arthritis) were tested for PR3-ANCA and MPO-ANCA with the EliA assay (Pharmacia Diagnostics, Freiburg, Germany). For comparison purposes, the same sera were also tested by indirect immunofluorescence, another direct immunometric assay (Varelisa, Pharmacia Diagnostics) and a capture PR3-ANCA (Wieslab AB, Lund, Sweden) method. Both the EliA PR3-ANCA and MPO-ANCA assays showed between- and within-assay precision of <10%. The dilution test gave straight lines (r2=0.998) for both antibody assays. The recovery ranged from 97.9% to 102.7% for PR3-ANCA and from 84.9% to 91.4% for MPO-ANCA. There was a high positive correlation between the EliA and Varelisa methods for quantitative detection of MPO-ANCA levels (r2=0.949) and a lower correlation for PR3-ANCA (r2=0.771). Conversely, poor correlation was observed between EliA PR3-ANCA and capture PR3-ANCA (r2=0.537). The overall sensitivity and specificity of EliA PR3-ANCA and MPO-ANCA for the vasculitides considered in this study were 82.7% and 97.2%, respectively, with a positive predictive value of 96.6% and a negative predictive value of 84.9%. Comparison of the results obtained with the indirect immunofluorescence, Varelisa and capture PR3-ANCA methods showed that the indirect immunofluorescence assay is the most sensitive method for the diagnosis of vasculitis (88.5%), but the least specific (94.3%); the EliA method is slightly more specific (97.2%) than the Varelisa method (95.7%), and also slightly more sensitive (82.7% vs. 80.8%). Capture PR3-ANCA proved to be the most sensitive method for detection of anti proteinase 3 antibodies in WG (89.7% vs. 86.2% EliA and 79.3% Varelisa). In conclusion, the EliA MPO-ANCA and PR3-ANCA methods provide good diagnostic accuracy and excellent analytical accuracy, which, in association with the practicality of the automated EliA system, make this method a useful tool for the diagnosis of ANCA-associated vasculitides. PMID- 15552277 TI - A particle-enhanced turbidimetric immunoassay for quantitative determination of orosomucoid in urine: development, validation and reference values. AB - Increased urinary orosomucoid excretion rate (UOER) is an independent predictor of cardiovascular and all-cause mortality in patients with type 2 diabetes, as demonstrated by a conventional, immunoturbidimetric method. We wanted to optimize the method by developing a fully automated, particle-enhanced turbidimetric (PET) immunoassay with a lower detection limit, to allow assessment of orosomucoid in urine in healthy individuals and patients. A micro-particle-based immunoreagent was prepared for a PET immunoassay. The calibration was traceable to the certified reference material (CRM 470) for specific human serum proteins. We studied 69 healthy adults (28 men and 41 women) to establish reference values for the new assay. The detection limit of orosomucoid in urine was found to be 0.05 mg/l, about 20 times lower than for the conventional assay. Within-run imprecision [CV%, (level)] was 6.7% (0.23 mg/l), 1.0% (1.08 mg/l) and 1.0% (4.69 mg/l). Total imprecision [CV%, (level)] was 10.4% (0.23 mg/l), 3.9% (1.08 mg/l) and 3.4% (4.69 mg/l). Reference values [median (2.5-97.5 percentiles)] for UOER were 0.36 (0.07-2.04) microg/min and for urinary orosomucoid/creatinine ratio 0.04 (0.009-0.17) mg/mmol. We describe a fully automated, transferable, sufficiently precise, high-sensitivity assay for orosomucoid in urine and present reference values traceable to CRM 470. PMID- 15552278 TI - Analytical performance and clinical results of a fully automated MEIA system for brain natriuretic peptide assay: comparison with a point of care testing method. AB - The aim of this study was to evaluate the analytical performance of a recently available immunoassay for brain natriuretic peptide (BNP), based on microparticle enzyme immunoassay (MEIA, AxSYM System, Abbott Laboratories), whose analytical characteristics and clinical results were compared with those of a point of care testing (POCT) method (TRIAGE system, Biosite Diagnostics). The within-run and total imprecision of the MEIA system were 18.4% and 19.8% at 21 ng/l, 8.0% and 14.8% at 183 ng/l, and 5.7% and 14.0% at 319 ng/l, respectively. The detection limit of the MEIA system was tested by repeatedly measuring (n=20) the 0 calibrator in four different runs; a mean +3 SD value of 5.6+/-4.8 ng/l (range 1.8-12.6 ng/l) was obtained. A close linear relationship (MEIA= -22.5+/-1.71 POCT method, R=0.950, n=296) was found (BNP concentration: 5-5500 ng/l), with a significant bias (mean difference: 164.8 ng/l, p<0.0001). Mean BNP concentration measured in 94 reference subjects (57 women and 37 men; mean age 43.5+/-14.0 years) was higher with MEIA than POCT, (25.9+/-32.7 ng/l vs. 11.7+/-8.9 ng/l, p<0.0001). The same trend was observed also in 202 cardiac patients (620.6+/ 1082.2 ng/l vs. 386.1+/-594.5 ng/l, p<0.0001). Our data suggest that MEIA and POCT have quite similar analytical performance but different clinical results. Then, different reference values, as well as cut-off values, should be taken into account for the clinical use of these two immunoassays. PMID- 15552279 TI - Multicentre performance evaluation of the E170 module for modular analytics. AB - The E170 module was evaluated at 13 sites in an international multicentre study. The objective of the study was to assess the analytical performance of 49 analytes, and to collect feedback on the system's reliability and practicability. The typical, within-run coefficients of variation (CVs) for most of the quantitative assays ranged between 1 and 2% while a range of 2-4% was achieved with the infectious disease methods. Total precision CVs were found to be within the manufacturer's expected performance ranges, demonstrating good concordance of the system's measuring channels and a high reproducibility during the 2-4-week trial period. The functional sensitivity of 11 selected assays met the clinical requirements (e.g., thyreotroponin (TSH) 0.008 mU/l, troponin T 0.02 microg/l, total prostate-specific antigen (PSA) 0.03 microg/l). The E170 showed no drift during an 8-hour period and no relevant reagent carryover. Accuracy was confirmed by ring trial experiments and method comparisons vs. Elecsys 2010. The reliability and practicability of the system's hardware and software met with, or even exceeded, the evaluator's requirements. Workflow studies showed that E170 can cover the combined workload of various routine analysers in a variety of laboratory environment. Throughput and sample processing time requirements were achieved while personnel 'hands-on-time' could be reduced. PMID- 15552280 TI - Beta-thalassaemia--a major health problem in Pakistan. PMID- 15552281 TI - Allogeneic bone marrow transplantation in beta-thalassaemia--single centre study. AB - OBJECTIVE: To evaluate outcome of allogeneic BMT in beta-Thalassaemia at the Armed Forces Bone Marrow Transplant Centre, Rawalpindi, Pakistan from August 2001 to November 2003. METHODS: Nineteen patients with beta-Thalassaemia underwent allogeneic BMT/PBSC transplantation from HLA identical sibling donors. Patients were classified in three groups according to Pesaro (Italy) risk classification. Class-I (n = 9) and Class-II (n = 7) patients received conditioning with busulphan/cyclophosphamide, whereas Class-III (n = 3) patient received conditioning with hydroxyurea, azathioprine, fludarabine, along with Bu14 / Cy 200. Cyclosporine, prednisolone and methotrexate were given for GvHD prophylaxis. Stem cells dose infused was >4.0 x 10(8)/kg body weight of the patient. RESULTS: Engraftment was achieved in all Class-I patients, whereas in Class-II and Class III , graft rejection was observed in one patient from each class. Median time to achieve absolute neutrophil recovery (> 0.5 x 10(9)/l) was 13 days, platelet count (> 20 x 10(9)/1) was 15 days and reticulocyte count (>0.5%) was 15 days. Acute GvHD was observed in 15 patients. One patient developed grade IV GvHD (liver and skin) and died within 30 days post BMT. Post transplant infectious complications were pseudomonas septicemia, disseminated fungal infection, CMV pneumonia and tuberculosis. Three patients died of these complications during post transplant period (31-90 days). Median stay in hospital was 25 days. CONCLUSION: Allogeneic BMT is the only curative therapy for beta-Thalassaemia patients, however the success rate can be increased if the patients are selected carefully and transplanted at an early age. PMID- 15552282 TI - The pregnancy rates with intrauterine insemination (IUI) in superovulated cycles employing different protocols (clomiphen citrate (CC), human menopausal gonadotropin (HMG) and HMG+CC) and in natural ovulatory cycle. AB - OBJECTIVE: To compare the result of IUI in infertile couples with different protocols of induction ovulation. METHODS: In a retrospective study, 209 infertile couples with different diagnosis (unexplained, male factor, endometriosis, tubal disease, ovulatory dysfunction and multifactorial infertility) were subjected to different protocol of induction ovulation: 50-100 mg CC in day 2-6, 50 mg CC in day 2-6 + 2 amp HMG in day 5, 7, 9, 11, and 2 amp HMG per day. Natural ovulatory cycle + IUI was used for sperm stored patients. 441 consecutive cycles of IUI was performed 36-40 hours after HCG injection. The data were analysed with student T-test and Mann-Whitney test. The significance was defined as P<0.005. RESULTS: Thirty one pregnancies (7% per cycle, 15% per patient) occurred. One pregnancy occurred (pregnancy per cycle was 2% and per patient was 12%) in 8 patients undergoing 37 cycles of IUI with natural ovulation. The result with CC in 27 patients undergoing 41 cycles IUI was 2 pregnancies (4% per cycle, 7% per patient). In 129 patients receiving 283 cycles of IUI with CC+HMG 21 pregnancies occurred (7% per cycle, 16% per patient). In 35 patients receiving 80 cycles of IUI with HMG 8 pregnancies occurred (9% per cycle, 23% per patient). CONCLUSION: The method chosen for ovulation induction had a critical bearing on the success of IUI. The result of IUI will be better by using induction ovulation compared to natural ovulatory cycle. In our programme the combined use of HMG+IUI yielded a higher rate of pregnancy rate compared with CC+IUI, CC+HMG+IUI and natural ovulatory cycle+IUI. PMID- 15552283 TI - Prevalence of hypertension in a low income settlement of Karachi, Pakistan. AB - OBJECTIVE: To assess the prevalence of hypertension in a low-income community in Karachi. METHODS: Cross sectional survey, through multi-stage sampling of 857 adult subjects in 405 households was conducted between April and September 2002. RESULTS: The overall prevalence of hypertension was 26% (95% C.I. 23, 29), the prevalence among males (34%) was higher than females (24%). The mean age of participants was 35 +/- 14 years, the prevalence of hypertension increased with age. Proportionately there were more cases of hypertension among male participants over 35 years of age as compared to female participants of the same age (p < or = 0.001). Fifty-eight percent of hypertensives were unaware of their hypertension. None of the hypertensive subjects who were aware of their condition had blood pressure under 140/90 mmHg. Stratified analysis revealed significant risk factors for hypertension. Hypertension was 1.7 (OR 95% C.I. 1.14, 2.42) times more common among males then females. Males were 1.7 (OR 95% C.I. 1.06, 2.6) times less likely to have been aware of their hypertension status. Age analysis revealed that the prevalence of hypertension increased with age and hypertensive subjects were 5.6 (OR 95% C.I. 3.9, 8.1) times more likely to be over 35 years of age. CONCLUSION: There is a need to control hypertension and prevent its complications through effective community interventions. The survey results indicate high prevalence and poor control of hypertension in the community, but more research is required to understand the reasons behind this phenomena. PMID- 15552284 TI - Profile of children with congenital adrenal hyperplasia--a hospital study. AB - OBJECTIVE: To collect baseline information on congenital adrenal hyperplasia (CAH) and to identify relevant issues specific to this disease in Pakistan. METHODS: A retrospective analysis of medical records of pediatric patients registered for serum 17 hydroxyprogesterone (17-OHP) measurement and documented to have CAH in the period 1987 to 1998 was carried out at The Aga Khan University, Karachi (AKU). The clinical notes were reviewed for documentation of CAH as the diagnosis. RESULTS: Of the 152 children registered for 17-OHP testing, sixty-three were diagnosed with CAH. Salt wasting, simple virilization and non classical CAH was found in 40 (63%), 18 (29.0%) and 5 (8.0%) patients respectively. Twenty-one (33.9%) patients were incorrectly assigned sex and of these, 20 (32.2%) patients were females who were either considered males or just not assigned gender. Parental consanguinity was found in 33 (52.3%) cases. No case had a history of similar features in either parent but in 19 (30.6%) cases similar features were present in siblings. Sixteen cases (25.4%) had a history of sibling death in the neonatal period and 7 had a history of sibling death in infancy. Maternal obstetric histories identified 3 (4.8%) cases with a history of still birth(s) and 4 (6.4%) with a history of abortion(s). CONCLUSION: Children with CAH should be diagnosed early as a rational and judicious choice of sex assignment is a critical aspect of treatment. The high rate of consanguinity emphasized the need to establish the true incidence of the defect in Pakistani population. PMID- 15552285 TI - Burden of gynaecological disease in a tertiary hospital: two years audit of outpatient department at PIMS. AB - OBJECTIVE: To estimate the pattern of gynaecological morbidity in women seeking services. METHODS: All women seeking advice for gynaecological problems in the outpatients department of Obstetrics and Gynaecology, Pakistan Institute of Medical Sciences Islamabad, from January 1997 to December 1998 were included. RESULTS: The total reproductive morbidity presenting at the outpatients department was 29,196. Of these 18,289 (62.7%) presented with obstetric morbidity and 10,851 (37.2%) with gynaecological morbidity. Menstrual irregularity was the commonest gynaecological problem encountered (41.1%). The rest included reproductive tract infections (27.8%), subfertility (18.2%), urogynaecological problems (5.3%), menopausal symptoms (4%), benign genital tract tumors (2. 8%), gynaecological malignancies (0.7%) and others (0.1%, including sexual dysfunction, congenital malformations and genital tract injuries) CONCLUSION: A number of women are seeking help for the varying severity of gynaecological morbidity, ranging from mild discomfort to overt disability. The major gnaecological morbidities identified are menstrual irregularities and reproductive tract infections. This study highlights the need for addressing and prioritising resources towards these women health issues which affect women's ability to fulfill a wide range of diverse roles. PMID- 15552286 TI - Management of blunt renal trauma: a profile of 65 patients. AB - OBJECTIVE: To outline the pattern and trends in the management of patients of renal trauma at the Department of Urology and Renal Transplantation, Jinnah Hospital, Lahore, from January 1998 to June 2003. METHODS: We retrospectively reviewed 65 patients identified with blunt renal trauma. RESULTS: Of 65 patients of renal trauma, injuries were graded I to V in 18 (27%), 9 (14%), 13 (20%), 16 (24%) and 9 (14%) respectively. Nephrectomy was done in all patients of Grade V and in 3 patients of Grade IV injury. Two patients in each group of grade IV and V died of associated injury. Rest of the patients were treated conservatively with blood transfusion, parenteral fluid, antibiotics, analgesics and bed rest. Of 53 patients treated conservatively two (4%) developed fever and eight (15%) hypertension. Two (4%) patients of grade III underwent delayed exploration for perinephric collection. CONCLUSION: Conservative management of blunt renal trauma without associated abdominal injury is feasible in patients who are hemodynamically stable at presentation. PMID- 15552287 TI - Prevalence of depression and the associated risks factors among adult women in a fishing community. AB - OBJECTIVE: To determine prevalence of depression and its associated risk factors with depression among adult women in a fishing community. METHODS: This study was conducted in Sualehabad, a small community with a population of 5000 of Manora Island in two stages. In first stage door to door survey was conducted and about 1200 women were screened for depression by using Mini International Neuropsychiatric Interview by Sheehan which was supplemented by ICD-10 Diagnostic Classification. In order to determine the associated risk factors, 91 controls were selected among the non-depressed women randomly from the same community with the 91 identified cases of depression. In second stage, a semi-structured questionnaire was administered to both the cases and controls, which was then subjected to reliability and validity exercises. The total duration of the study was six months. RESULTS: The point prevalence of depressive disorder was 7.5%. Only 13% reported treatment from government facility and 14% reported previous consultation with a psychiatrist. Sixteen percent of women were aware about mental health facility available in the area. Twenty seven percent reported that they got relief from alternative treatment. The characteristics that demonstrated a statistically significant association related to risk factors were increasing age, being married, more than four children in family, illiteracy and financial difficulties at home. There was a significant association observed in reporting social, behavioral and relational factors related to depression by cases as compared to control group. CONCLUSION: The prevalence of depressive disorder among adult women of the fishing community was 7.5% which is at a high magnitude keeping in view the prevalence rate of the country and the global estimates. There is a need for more community based studies in order to assess the magnitude of depression which is posing one of the major public health problems. PMID- 15552288 TI - Clinical presentation of thyroid cancer patients in Pakistan--AKUH experience. AB - OBJECTIVE: To asses the clinical presentation of patients with thyroid cancer, their preferred diagnostic work-up and preferred surgical procedure. METHODS: A retrospective chart review of patients that underwent partial or total thyroidectomy for thyroid cancer at AKUH between 1989 and 2002. SPSS was used to analyze data. Pearson's chi square and likelihood ratios used to calculate tests of significance. RESULTS: A total of 97 patients were included in this review. Solitary thyroid nodule was the initial presentation in three. Majority (58.7%) presented with "neck swelling" or enlarging goiter. One out of every 3 patients showed evidence of lymph node involvement and 20% had distant metastases. The aggressiveness of the disease did not correlate with age, sex or histological subtype. There was no correlation between the size of the tumor and presence of local or distant metastases. CONCLUSION: Thyroid cancer is a more aggressive disease in Pakistan, with majority of patients presenting with multinodular goiters, and a significant number have lymph node metastases. A higher degree of vigilance and a lower threshold for fine needle aspiration (FNA) is needed while evaluating patients with thyroid goiter. PMID- 15552289 TI - Quantitative and qualitative assessment of biomedical publications from Iran, Pakistan and Egypt through their impact factor. PMID- 15552290 TI - Assessment of physicians understanding of asthma guidelines in a tertiary care hospital. PMID- 15552291 TI - Cystic breast lymphangioma. PMID- 15552292 TI - Pleomorphic liposarcoma in a ten year old child. PMID- 15552293 TI - Attitudes and practices of postgraduate medical trainees towards research--a snapshot from Faisalabad. AB - OBJECTIVE: To assess the attitudes and practices of postgraduate medical trainees towards research. METHODS: It was a self-administered questionnaire based cross sectional survey conducted on 55 conveniently selected trainees in Allied Hospital, Faisalabad. RESULTS: Only 11 trainees read journals monthly, seven had written an article for a journal, 51 regarded reading literature important, 39 intended to engage in future research and 37 said they received inappropriate research training. The major reasons cited for poor research activity in Pakistan were poor research training and awareness. CONCLUSION: Though the attitudes towards research were positive, they were deficient practically in terms of reading and writing literature. There is an immediate need to improve research training in our educational institutes to facilitate the development of the local literature both in terms of research utilization and production PMID- 15552295 TI - Abstracts of case reports. PMID- 15552296 TI - Multidisciplinary task force tackles Chicago distemper outbreak. Questions raised about vaccinations. PMID- 15552297 TI - The catch-22 in regulating the slaughter industry. USDA encourages improvements in animal welfare at slaughterhouses. PMID- 15552298 TI - Small-Business Summit marks legislative successes, goals. Health care costs, frivolous lawsuits key concerns. PMID- 15552299 TI - Mammal mapping projects to move forward. NIH changes procedure for choosing sequencing targets. PMID- 15552300 TI - A vision of a no-kill future. PMID- 15552301 TI - Finds evidence-based conclusions missing from study. PMID- 15552302 TI - Thoughts on legal classification of pets as sentient property. PMID- 15552303 TI - What is your diagnosis? Calcium crystal deposition disease (chondrocalcinosis). PMID- 15552304 TI - What is your diagnosis? Horner syndrome. PMID- 15552305 TI - Animal behavior case of the month. Two dogs were evaluated for episodic bouts of bizarre behavior. PMID- 15552306 TI - Environmental surveillance for Salmonella enterica in a veterinary teaching hospital. AB - OBJECTIVE: To evaluate the extent of environmental contamination with Salmonella enterica in a veterinary teaching hospital. DESIGN: Longitudinal study. SAMPLES: Environmental samples obtained from 69 representative locations within a veterinary teaching hospital by use of a commercially available electrostatic wipe. PROCEDURE: Environmental samples were obtained for bacteriologic culture, and antimicrobial susceptibility testing was performed on each environmental isolate. Environmental isolates were compared with isolates obtained from animals during the same period to investigate potential sources of environmental contamination. RESULTS: 54 S. enterica isolates were recovered from 452 (11.9%) cultured environmental samples. Five different serotypes were recovered; the most common serotypes were S. Newport and S. Agona. Within the 5 serotypes recovered, 10 distinguishable phenotypes were identified by use of serotype and antimicrobial susceptibility patterns. Of the environmental isolates, 41 of 54 (75.9%) could be matched to phenotypes of isolates obtained from animal submissions in the month prior to collection of environmental samples. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that environments in veterinary hospitals can be frequently contaminated with S. enterica near where infected animals are managed and fecal specimens containing S. enterica are processed for culture in a diagnostic laboratory. Bacteriologic culture of environmental samples collected with electrostatic wipes is an effective means of detecting contamination in a veterinary hospital environment and may be beneficial as part of surveillance activities for other veterinary and animal rearing facilities. PMID- 15552307 TI - Understanding issues and solutions for unowned, free-roaming cat populations. PMID- 15552308 TI - Humane strategies for controlling feral cat populations. PMID- 15552309 TI - Analyzing approaches to feral cat management--one size does not fit all. PMID- 15552310 TI - Professional, ethical, and legal dilemmas of trap-neuter-release. PMID- 15552311 TI - Trap-neuter-release programs: the reality and the impacts. PMID- 15552312 TI - The welfare of feral cats and wildlife. PMID- 15552313 TI - Laboratory quality considerations for veterinary practitioners. PMID- 15552314 TI - Seroprevalences of antibodies against Bartonella henselae and Toxoplasma gondii and fecal shedding of Cryptosporidium spp, Giardia spp, and Toxocara cati in feral and pet domestic cats. AB - OBJECTIVE: To compare seroprevalences of antibodies against Bartonella henselae and Toxoplasma gondii and fecal shedding of Cryptosporidium spp, Giardia spp, and Toxocara cati in feral and pet domestic cats. DESIGN: Prospective cross-sectional serologic and coprologic survey. ANIMALS: 100 feral cats and 76 pet domestic cats from Randolph County, NC. PROCEDURE: Blood and fecal samples were collected and tested. RESULTS: Percentages of feral cats seropositive for antibodies against B. henselae and T. gondii (93% and 63%, respectively) were significantly higher than percentages of pet cats (75% and 34%). Percentages of feral and pet cats with Cryptosporidium spp (7% of feral cats; 6% of pet cats), Giardia spp (6% of feral cats; 5% of pet cats), and T. cati ova (21% of feral cats; 18% of pet cats) in their feces were not significantly different between populations. Results of CBCs and serum biochemical analyses were not significantly different between feral and pet cats, except that feral cats had a significantly lower median PCV and significantly higher median neutrophil count. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that feral and pet cats had similar baseline health status, as reflected by results of hematologic and serum biochemical testing and similar prevalences of infection with Cryptosporidium spp, Giardia spp, and T. cati. Feral cats did have higher seroprevalences of antibodies against B. henselae and T. gondii than did pet cats, but this likely was related to greater exposure to vectors of these organisms. PMID- 15552315 TI - Reproductive capacity of free-roaming domestic cats and kitten survival rate. AB - OBJECTIVE: To determine reproductive capacity of naturally breeding free-roaming domestic cats and kitten survival rate. DESIGN: Prospective cohort and retrospective cross-sectional study. ANIMALS: 2,332 female cats brought to a trap neuter-return clinic for neutering and 71 female cats and 171 kittens comprising 50 litters from a cohort study of feral cats in managed colonies. PROCEDURE: Data collected for all cats included pregnancy, lactation, and estrus status and number of fetuses for pregnant cats. Additional data collected for feral cats in managed colonies included numbers of litters per year and kittens per litter, date of birth, kitten survival rate, and causes of death. RESULTS: Pregnant cats were observed in all months of the year, but the percentage of cats found to be pregnant was highest in March, April, and May. Cats produced a mean of 1.4 litters/y, with a median of 3 kittens/litter (range, 1 to 6). Overall, 127 of 169 (75%) kittens died or disappeared before 6 months of age. Trauma was the most common cause of death. CONCLUSIONS AND CLINICAL RELEVANCE: Results illustrate the high reproductive capacity of free-roaming domestic cats. Realistic estimates of the reproductive capacity of female cats may be useful in assessing the effectiveness of population control strategies. PMID- 15552316 TI - Time and financial costs of programs for live trapping feral cats. AB - OBJECTIVE: To determine the time and financial costs of programs for live trapping feral cats and determine whether allowing cats to become acclimated to the traps improved trapping effectiveness. DESIGN: Prospective cohort study. ANIMALS: 107 feral cats in 9 colonies. PROCEDURE: 15 traps were set at each colony for 5 consecutive nights, and 5 traps were then set per night until trapping was complete. In 4 colonies, traps were immediately baited and set; in the remaining 5 colonies, traps were left open and cats were fed in the traps for 3 days prior to the initiation of trapping. Costs for bait and labor were calculated, and trapping effort and efficiency were assessed. RESULTS: Mean +/- SD overall trapping effort (ie, number of trap-nights until at least 90% of the cats in the colony had been captured or until no more than 1 cat remained untrapped) was 8.9 +/- 3.9 trap-nights per cat captured. Mean overall trapping efficiency (ie, percentage of cats captured per colony) was 98.0 +/- 4.0%. There were no significant differences in trapping effort or efficiency between colonies that were provided an acclimation period and colonies that were not. Overall trapping costs were significantly higher for colonies provided an acclimation period. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that these live trapping protocols were effective. Feeding cats their regular diets in the traps for 3 days prior to the initiation of trapping did not have a significant effect on trapping effort or efficiency in the present study but was associated with significant increases in trapping costs. PMID- 15552317 TI - Antiplatelet effects and pharmacodynamics of clopidogrel in cats. AB - OBJECTIVE: To evaluate antiplatelet effects and pharmacodynamics of clopidogrel in cats. DESIGN: Original study. ANIMALS: 5 purpose-bred domestic cats. PROCEDURE: Clopidogrel was administered at dosages of 75 mg, p.o., every 24 hours for 10 days; 37.5 mg, p.o., every 24 hours for 10 days; and 18.75 mg, p.o., every 24 hours for 7 days. In all cats, treatments were administered in this order, with at least 2 weeks between treatments. Platelet aggregation in response to ADP and collagen and oral mucosal bleeding times (OMBTs) were measured before and 3, 7, and 10 days (75 and 37.5 mg) or 7 days (18.75 mg) after initiation of drug administration. Serotonin concentration in plasma following stimulation of platelets with ADP or collagen was measured before and on the last day of drug administration. Platelet aggregation, OMBT, and serotonin concentration were evaluated at various times after drug administration was discontinued to determine when drug effects were lost. RESULTS: For all 3 dosages, platelet aggregation in response to ADP platelet aggregation in response to collagen, and serotonin concentration were significantly reduced and OMBT was significantly increased at all measurement times during drug administration periods. All values returned to baseline values by 7 days after drug administration was discontinued. No significant differences were identified between doses. None of the cats developed adverse effects associated with drug administration. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that administration of clopidogrel at dosages ranging from 18.75 to 75 mg, p.o., every 24 hours, results in significant antiplatelet effects in cats. PMID- 15552318 TI - Use of cisplatin, 5-fluorouracil, and second-look laparotomy for the management of gastrointestinal adenocarcinoma in three dogs. AB - Three dogs with advanced-stage adenocarcinoma of the gastrointestinal tract were treated by use of resection, adjuvant chemotherapy with cisplatin and 5 fluorouracil, and second-look laparotomy (SLL). In combination with histologic examination of biopsy specimens obtained during the procedure, SLL is the most accurate diagnostic procedure for identification of residual or recurrent microscopic or macroscopic abdominal neoplasia; however, to the authors' knowledge, there are no reports of its clinical use in the field of veterinary oncology. This lack of clinical use in animals is likely because of factors such as cost, procedure-associated risks perceived by the owners and veterinarians, lack of data to define proper clinical application, and, perhaps to some degree, an entrenched belief that treatment of advanced stage cancer in animals is inappropriate. Nevertheless, the use of SLL should be considered for evaluation of abdominal tumors or intra-abdominal metastases in dogs that appear to be in complete clinical remission near or at the anticipated completion of chemotherapy (especially if effective second-line chemotherapy protocols are available) or when secondary cytoreduction might be beneficial. PMID- 15552319 TI - Surgical management of gallbladder mucoceles in dogs: 22 cases (1999-2003). AB - OBJECTIVES: To describe preoperative, surgical, and postoperative findings and determine prognostic indicators and treatment recommendations in dogs treated surgically for gallbladder mucocele. DESIGN: Retrospective study. ANIMALS: 22 client-owned dogs. PROCEDURES: Medical records of dogs with gallbladder mucoceles that were treated surgically were reviewed. History, clinical signs, results of selected clinicopathologic analyses and abdominal ultrasonography, surgical procedure performed, results of histologic examination of a liver biopsy specimen, and survival time were recorded. Follow-up information was obtained via telephone interview with owners and referring veterinarians. RESULTS: Dogs were 7 to 15 years of age and had non-specific clinical signs (vomiting, anorexia, and lethargy). Physical examination findings included icterus, signs of depression, and signs of discomfort on palpation of the abdomen. Sixteen dogs had a definitive diagnosis and 6 dogs were strongly suspected of having a gallbladder mucocele on the basis of results of abdominal ultrasonography. Fifteen dogs survived after surgery; 3 of these dogs had bile-induced peritonitis, and 4 had pancreatitis. One dog was euthanatized as a result of severe pancreatitis, and 1 was euthanatized because of acute renal failure; 5 dogs died as a result of pancreatitis, cholecystitis, or bile-induced peritonitis. Hepatic abnormalities were detected histologically in all dogs. CONCLUSIONS AND CLINICAL RELEVANCE: No predictors of survival were identified. No associations between outcome of surgical treatment (survival vs nonsurvival) and preoperative findings, biliary rupture, surgical procedure performed, results of histologic examination of the liver, or development of pancreatitis were found. Cholecystoduodenostomy and cholecystectomy appear to be acceptable treatments for gallbladder mucocele. PMID- 15552320 TI - Surgical management of compound odontoma in two horses. AB - Two horses were admitted for evaluation of mandibular swelling (horse 1) or maxillary distortion (horse 2). Both horses had radiographic evidence of tumors of dental origin that had the appearance of a compound odontoma. Extensive surgical resection was performed for treatment. Horse 1 was treated with 1-stage surgical resection, but an iatrogenic fracture occurred during surgery, which was managed successfully with a type I external fixator and extraoral alimentation. Horse 2 was treated in multiple stages to remove all portions of the tumor. To manage an extensive orosinal fistula, a custom-designed dental bridge was constructed to occlude the fistula. For both horses, the histopathologic diagnosis was compound odontoma. Compound odontomas are benign, locally expansive tumors of dental origin. Compound odontomas can be treated successfully; however, multiple surgeries may be necessary. PMID- 15552322 TI - Ethical limits in human subjects research. PMID- 15552321 TI - Evaluation of magnetic resonance imaging for detection of internal tumors in green turtles with cutaneous fibropapillomatosis. AB - OBJECTIVE: To describe the gross cross-sectional anatomy of green turtles (Chelonia mydas) and evaluate magnetic resonance imaging (MRI) for detection of internal tumors in green turtles with cutaneous fibropapillomatosis. DESIGN: Prospective study. ANIMALS: 3 dead green turtles, 1 healthy green turtle, and 8 green turtles with cutaneous fibropapillomatosis. PROCEDURES: Gross cross sectional anatomy of a dead turtle was described. Each live turtle underwent a complete physical examination, and dorsoventral whole-body survey radiographic views were obtained. Magnetic resonance imaging was performed in dorsal and transverse planes. Radiographs and magnetic resonance images were examined for evidence of internal nodules. Results were compared with necropsy findings in 5 of 8 turtles. RESULTS: Nodules in the lungs of 2 turtles were detected via radiography, whereas pulmonary nodules were detected in 5 turtles via MRI. No other visceral nodules were detected via radiography; however, masses in the stomach and adjacent to the bladder and kidneys were detected in 1 turtle via MRI. Other extrapulmonary abnormalities observed at necropsy were not detected on MR images. CONCLUSIONS AND CLINICAL RELEVANCE: MRI may be valuable for detection of internal tumors in green turtles with cutaneous fibropapillomatosis. Nodules were more apparent in the lungs than in other organs. Results of MRI may serve as prognostic indicators for sea turtles undergoing assessment, treatment, and rehabilitation. Clinical application may be limited by cost and availability of MRI technology. PMID- 15552323 TI - The ethics of biobanks. PMID- 15552324 TI - Committee for Oversight of Research Involving the Dead (CORID): insights from the first year. PMID- 15552325 TI - The involuntary research subject. PMID- 15552326 TI - Informed consent in pediatric research. PMID- 15552327 TI - The poster child for the need for central review of research protocols: the Children's Oncology Group. PMID- 15552328 TI - Inconsistent regulatory protection under the U.S. Common Rule. PMID- 15552329 TI - Placebo controlled trials: restrictions, not prohibitions. PMID- 15552330 TI - Ethical dilemmas in retrospective studies on genital surgery in the treatment of intersexual infants. PMID- 15552331 TI - HIV/AIDS clinical research, and the claims of beneficence, justice, and integrity. PMID- 15552332 TI - CQ sources/bibliography. PMID- 15552333 TI - Genetic testing after breast cancer diagnosis: implications for physician-patient communications. PMID- 15552334 TI - A diagnosis of undue influence: Congressman Henry Waxman on science and politics. PMID- 15552335 TI - Two perspectives. PMID- 15552336 TI - Applying critical thinking skills to character education and values clarification with students who are deaf or hard of hearing. AB - Students who are deaf or hard of hearing must learn to think critically. Character education (CE) refers to the effort to teach basic values and moral reasoning (Doyle & Ponder, 1977). Values clarification (VC) is the process of examining one's basic values and moral reasoning (Rokeach, 1973). Character education and values clarification as subject matter foster the development of critical thinking (CT), a tool used both to develop and to modify values and moral reasoning. These three areas mutually support one another. The development of a set of values and their underlying moral reasoning is the foundation for thinking critically about values. The authors examine the components of critical thinking, character education, and values clarification, summarize the literature, and provide a template for appropriate lesson plans. They also describe strategies that promote the development of critical thinking, character education, and values clarification. PMID- 15552337 TI - Factors involved in the ineffective dissemination of sexuality information to individuals who are deaf or hard of hearing. AB - The last 40 years of literature pertaining to sexuality and deaf individuals are reviewed. Current research, which establishes that people who are deaf do not have adequate information on sexuality issues, is examined, as well as some of the factors that play a role in the ineffective dissemination of sexuality information to this population. Parents, education (both in a historical and a contemporary light), peers, and the very acquisition of language are examined with regard to their contextualized part in the process of knowledge sharing. Historical paradigms are placed within Griffiths's "mythconceptions" framework (as discussed in Watson, 2002) in an effort to determine possible causative factors relating to deaf people's insufficient knowledge regarding sexuality. PMID- 15552338 TI - Deaf children in regular classrooms: a sociocultural approach to a Brazillan experience. AB - Inclusion of deaf children in regular classrooms is often described as unsuccessful. The present article shows how communicative and metacommunicative strategies used in teacher(s)-deaf students(s) interactions may facilitate inclusion. A fourth-grade classroom was investigated where a coteaching approach- a master teacher working with a teacher trained in Brazilian Sign Language (BSL)- was used. The class, 7 deaf and 19 hearing students, was selected because of the teacher dyad's effectiveness with these students. The teachers' interactive styles and strategies are highlighted, along with communicative and metacommunicative processes that occurred between them and the deaf students. The authors show that meanings are co-constructed not only through words or BSL but through nonverbal actions. Relational metacommunicative strategies make integration more effective and learning easier and more pleasant; therefore, dialogue with deaf children entails more than the mere use of words, either vocally or with signs. PMID- 15552339 TI - Comparing motor development of deaf children of deaf parents and deaf children of hearing parents. AB - Deaf children of Deaf parents perform better academically (Ritter-Brinton & Stewart, 1992), linguistically (Courtin, 2000; M. Harris, 2001; Vaccari & Marschark, 1997), and socially (Hadadian & Rose, 1991; M. Harris, 2001) than Deaf children of hearing parents. Twenty-nine Deaf children in residential schools were assessed to determine if a significant difference also exists in motor development between Deaf children with Deaf parents and Deaf children with hearing parents. In the locomotor area, 78.6% of Deaf children of Deaf parents and 73.3% of Deaf children of hearing parents reached or surpassed average performance levels. In regard to object control, 92.9% of Deaf children of Deaf parents and 93.3% of Deaf children of hearing parents reached or surpassed average performance levels. The study results show no significant difference between the motor development of Deaf children of Deaf parents and Deaf children of hearing parents. PMID- 15552341 TI - Attention-deficit hyperactivity disorder: setting the record straight. PMID- 15552340 TI - Preparing elementary education majors to teach science using an inquiry-based approach: the Full Option Science System. AB - Science education traditionally has received insufficient attention. As a literature review shows, teacher preparation in science will be best served by improvements in pedagogy and in the content of required undergraduate science courses. The American Association for the Advancement of Science (1993, 1995) and the National Research Council (1993, 1995) have addressed this need in advocating a "science for all" that is highly significant for diverse learners. The No Child Left Behind Act emphasizes that reform of teacher preparation is part of an urgent national commitment to bring high-quality teacher candidates into the classroom. The Gallaudet University undergraduate teacher education program has developed an inquiry-based course that emphasizes integration of the sciences. Acquisition of the Full Option Science System, and its adaptation to and integration into the course, has resulted in specific curricular changes and positive results. PMID- 15552342 TI - Intrauterine infections: a literature review. AB - During gestation, many microorganisms can infect the fetus, causing severe birth defects. Such organisms and the resulting clinical syndromes have been categorized as TORCH infections, a useful acronym referring to Toxoplasma gondii, other microorganisms (like syphilis), rubella virus, cytomegalovirus, and herpesviruses. Since dental patients typically report pregnancy or previous infections as part of their health history, and TORCH infections can manifest oral symptoms, the dentist is in a position to act as an educator and screener for these infections. This article reviews TORCH agents for dentists to help the clinician in educating pregnant patients about the risks these infections pose to the fetus. The authors also note oral symptoms related to these infections. PMID- 15552343 TI - Oral hygiene and oral symptoms among the elderly in long-term care. AB - Dental teams examined 1910 elderly adults living in long-term care settings (1358 institutionalized, 552 homebound) from all 19 counties in Norway to document oral hygiene, oral symptoms and ability to receive dental treatment. The Mucosal Plaque Score (MPS) was used to assess oral status. The MPS was significantly better in women than in men, in individuals with dentures than in those with any remaining teeth, and in people who were homebound rather than those who lived in institutions. The MPS did not differ between age groups or geographic regions. According to the Treatment Ability Index, nearly a quarter of the sample was able to receive comprehensive dental care. Reports of "any oral symptom" and "eating/chewing problems" decreased with age and were most prevalent for individuals who had dentures. The MPS had only slight impact on oral symptoms, chewing ability and dry mouth (p > 0.05). Dry mouth was found in 16.9% and was most prevalent in individuals with dentures. PMID- 15552344 TI - Prosthodontic management of older severe hypodontia patients in general dental practice: a case series. AB - Although a detailed understanding of the etiology of hypodontia is lacking, there is a need for awareness among dental professionals of the social and psychological consequences of severe hypodontia, as well as knowledge of those affected by hypodontia have ongoing dental treatment needs. Although, there may be a need for immediate referral of children to different specialists for optimal multidisciplinary treatment planning, most older patients with severe hypodontia can be treated by general practitioners. Practitioners therefore need to be aware of the ramifications of the condition and be capable of providing adequate oral health care for these patients and referring them for additional services when necessary. A case-report series is used to illustrate routine prosthodontic treatments that can be offered to older patients in general dental practice. PMID- 15552345 TI - Dental status and oral health quality of life in economically disadvantaged French adults. AB - Expansion of French health insurance coverage has increased funding for dental care for economically disadvantaged adults. This study aimed to measure clinical and self-perceived oral health, behaviors, and use of dental services by adults who were eligible for such coverage. The regional agency that gives administrative services for the health insurance funds provided a sample of 900 adults aged 35-44 years, insured through this program. We reached 805 of these adults by mail; of these 18% were surveyed and clinically examined. Self perceived oral health was measured by the Global Oral Health Assessment Index (GOHAI) and participants' attitudes to dental health, by questionnaire. Decayed and Missing teeth constituted 40% of the DMFT. Participants reported poor oral health (63%), and 79% perceived a need for care, although they used dental services infrequently and had poor knowledge of available services. Cost of care and number of carious teeth were important predictors of the GOHAI. PMID- 15552346 TI - Word length, phonemic, and visual similarity effects in poor and normal readers. AB - Serial order recall for visually and auditorily presented stimuli was examined in a group of 12-year-old poor readers and 7-year-old reading-age controls. With pictorial presentation, the poor readers showed a visual similarity effect, no word length effect, and a smaller phonemic similarity effect than that of controls. However, with visual presentation of printed words and with auditory presentation, poor readers showed word length and phonemic similarity effects of similar magnitude to that of controls. It is concluded that poor readers rely on visual information in tasks where the presented images are highly codable, and where verbal recoding is not obligatory, but that they will make use of phonological coding when the stimuli are not as easily codable visually in memory. PMID- 15552347 TI - Age differences in rereading. AB - Younger and older adults read a series of expository and narrative passages twice in order to answer comprehension questions. Reading time was used to index attentional allocation to word, textbase, and situation model processing and to assess shifts in the allocation policy from the first to the second reading. Older readers' comprehension was at least as good as that of younger readers. Analysis of reading times suggested that for both genres, older adults allocated more attention to situation model features than younger adults did on the first reading, whereas young and old allocated attention similarly to this level of representation on the second reading, suggesting that mature readers may give greater priority to situation model construction when first encountering text. Also, for both genres, older adults showed relatively less facilitation than the young in word-level processing in rereading, suggesting that representation at this level is not as firmly established during reading or decays more quickly for older readers. For narrative texts only, this pattern also obtained for textbase processing. Collectively, these data show that age equivalence in text comprehension at the molar level may be accomplished through different processing routes at the molecular level. PMID- 15552348 TI - The contribution to immediate serial recall of rehearsal, search speed, access to lexical memory, and phonological coding: an investigation at the construct level. AB - Rehearsal speed has traditionally been seen to be the prime determinant of individual differences in memory span. Recent studies, in the main using young children as the participant population, have suggested other contributors to span performance. In the present research, we used structural equation modeling to explore, at the construct level, individual differences in immediate serial recall with respect to rehearsal, search, phonological coding, and speed of access to lexical memory. We replicated standard short-term phenomena; we showed that the variables that influence children's span performance influence adult performance in the same way; and we showed that speed of access to lexical memory and facility with phonological codes appear to be more potent sources of individual differences in immediate memory than is either rehearsal speed or search factors. PMID- 15552349 TI - Valence of distractor words increases the effects of irrelevant speech on serial recall. AB - Participants memorized target words in silence or while ignoring neutral or valent (positive or negative) distractor words that could be either possessor relevant or other-relevant. Distractor words impaired recall performance, but valent distractor words caused more disruption than neutral distractors, and negative distractors caused more disruption than positive distractors. The results are problematic for explanations of the irrelevant speech effect within working memory models that do not specify an explicit role of attention in the maintenance of information for immediate serial recall. PMID- 15552350 TI - Visual phonology: the effects of orthographic consistency on different auditory word recognition tasks. AB - In this study, we investigated orthographic influences on spoken word recognition. The degree of spelling inconsistency was manipulated while rime phonology was held constant. Inconsistent words with subdominant spellings were processed more slowly than inconsistent words with dominant spellings. This graded consistency effect was obtained in three experiments. However, the effect was strongest in lexical decision, intermediate in rime detection, and weakest in auditory naming. We conclude that (1) orthographic consistency effects are not artifacts of phonological, phonetic, or phonotactic properties of the stimulus material; (2) orthographic effects can be found even when the error rate is extremely low, which rules out the possibility that they result from strategies used to reduce task difficulty; and (3) orthographic effects are not restricted to lexical decision. However, they are stronger in lexical decision than in other tasks. Overall, the study shows that learning about orthography alters the way we process spoken language. PMID- 15552351 TI - Can semantic relatedness explain the enhancement of memory for emotional words? AB - Memory for emotional items is often better than memory for neutral items. In three experiments, we examined whether this typical finding is due to the higher semantic relatedness inherent to emotional items, a confound in previous studies. We also controlled for other possible confounding variables, such as imagery. In Experiments 1 and 2, participants encoded lists of emotional and categorized neutral words equivalent in semantic relatedness, as well as lists of random neutral words with lower semantic relatedness. In Experiment 3, the lists were mixed, containing words from all the conditions. Surprise free recall was tested after a 40- to 55-min retention interval. Free recall of emotional words was better than that of random neutral words, replicating the classic effect. Importantly, categorized words were recalled better than random neutral words, and not worse than emotional words. These results emphasize the important role of semantic relatedness in the classic effect and suggest that organizational processes operate alongside arousal-related ones to enhance memory for emotional material. PMID- 15552352 TI - Phonology in the bilingual Stroop effect. AB - In a bilingual Stroop task, we examined between-language interference among proficient Japanese-English bilingual speakers. Participants named ink colors either in Japanese or in English. The Japanese color terms were either phonologically similar to (i.e., loan words) or dissimilar from (i.e., traditional color terms) English color terms. For both response languages, a significant between-language Stroop effect was found despite the orthographic dissimilarity between the languages. The magnitude of the between-language interference was larger with the phonologically similar terms. These findings implicate direct links connecting phonologically similar matching words in the lexicons of proficient bilingual speakers of dissimilar languages and imply that phonological processing in lexical access occurs even when the access is done unintentionally. PMID- 15552353 TI - Dissociating mere exposure and repetition priming as a function of word type. AB - The mere exposure effect is defined as enhanced attitude toward a stimulus that has been repeatedly exposed. Repetition priming is defined as facilitated processing of a previously exposed stimulus. We conducted a direct comparison between the two phenomena to test the assumption that the mere exposure effect represents an example of repetition priming. In two experiments, having studied a set of words or nonwords, participants were given a repetition priming task (perceptual identification) or one of two mere exposure (affective liking or preference judgment) tasks. Repetition priming was obtained for both words and nonwords, but only nonwords produced a mere exposure effect. This demonstrates a key boundary for observing the mere exposure effect, one not readily accommodated by a perceptual representation systems (Tulving & Schacter, 1990) account, which assumes that both phenomena should show some sensitivity to nonwords and words. PMID- 15552354 TI - Episodic memory change in late adulthood: generalizability across samples and performance indices. AB - Younger adults recall more information from episodic memory tasks than do older adults. Because longitudinal studies are rare and often incompatible, the extent of actual late-life memory change is not well established. We assemble two different longitudinal samples of normal older adults, each of which is tested twice at a 3-year interval, using a large battery of episodic memory indicators. Together, two-wave data from both the Victoria Longitudinal Study in Canada (n = 400) and the Kungsholmen Project in Sweden (n = 168) cover a 40-year span of adulthood, ranging from 54 to 94 years of age. Principal memory tasks include categorizable word lists, story recall, and random word lists, as well as indicators of cognitive support. Overall, an examination of performance on sets of common and complementary episodic tasks reveals that, for both samples, actual 3-year changes are modest and that, when decline occurs, it is gradual. The exception-greater decline for more supported tasks-suggests that these may be especially sensitive to late-life changes. PMID- 15552355 TI - Causes and constraints of the shift-to-easier-materials effect in the control of study. AB - The shift-to-easier-materials (STEM) effect occurs when individuals adopt a low performance goal and subsequently select to restudy more easier items of a list than the difficult ones. The causes and constraints of the STEM effect were investigated across four experiments in which participants first briefly studied and judged their learning of 30 paired associates. They were then instructed to obtain a low performance goal and were asked to select items for restudy. The STEM effect was present when items were presented for selection simultaneously, but when items were presented for selection sequentially, the participants instead selected the more difficult items. The presence and absence of STEM effects were linked to planning that was triggered by the simultaneous format and to difficulties in executing an appropriate plan under the sequential format, respectively. Moreover, the STEM effect was evident for individuals with high memory spans but not for those with low memory spans. These and other findings highlight the contribution of planning and capacity constraints to the control of study time. PMID- 15552356 TI - The episodic nature of involuntary autobiographical memories. AB - Involuntary autobiographical memories are conscious and unintended recollections of personal experiences. In Study 1, involuntary memories were compared with voluntary word-cued memories, both retrieved in naturalistic settings via a self paced procedure. The involuntary memories more frequently referred to specific episodes, came with more physical reaction, had more impact on mood, and dealt with more unusual and less positive events. Study 2 demonstrated that these differences were not due to differences between verbal and nonverbal cues, by using Francis Galton's "memory walk" as a nonverbal method to cue voluntary memories. In both studies, systematic differences were found between specific and nonspecific memories. The findings show that the way autobiographical memories are sampled greatly affects the findings and that involuntary retrieval more often provides access to memories of specific episodes and associated emotional states. PMID- 15552357 TI - Strengthening the activation of unconsciously activated memories. AB - Reading a word activates a set of associated words-for example, reading CAB activates car, driver, and other associates. This research was conducted to determine whether the unconscious activation of these associates adds activation to other associates in the same set. Words were studied, and recall was tested with cues comprised of associates directly activated by the target. On the basis of preexisting links, the associates serving as test cues also received strong (e.g., car) or weak (e.g., driver) additional inputs from other associates in the set. Cues receiving stronger inputs from other associates were more effective, indicating that unconsciously activated associates strengthen one another. This effect declined when conditions disrupted the retrieval of context features linked to the episode. Attending to another task during or after study or receiving implicit test instructions decreased the effects of input from other associates. Importantly, disruptions affected input from other associates more than direct input from the target, suggesting that the influence of context wanes as the links in the network become more distant. PMID- 15552358 TI - How generation affects source memory. AB - Generation effects (better memory for self-produced items than for provided items) typically occur in item memory. Jurica and Shimamura (1999) reported a negative generation effect in source memory, but their procedure did not test participants on the items they had generated. In Experiment 1, participants answered questions and read statements made by a face on a computer screen. The target word was unscrambled, or letters were filled in. Generation effects were found for target recall and source recognition (which person did which task). Experiment 2 extended these findings to a condition in which the external sources were two different faces. Generation had a positive effect on source memory, supporting an overlap in the underlying mechanisms of item and source memory. PMID- 15552359 TI - How similar is false recognition to veridical recognition objectively and subjectively? AB - Three recognition memory experiments were conducted using modified Deese-Roediger McDermott (DRM) and DRM paradigms. In Experiment 1, the reaction time (RT) of the false alarms to critical nonpresented words (false memory) was compared with the RT of hits to the critical presented words and with the RT of hits to the studied list words (true memory). The RT of the false alarms to the critical nonpresented words was significantly longer than that of the hits to the critical words and than that of the studied list words. In Experiment 2, in addition to RT, participants' confidence level was measured on a 4-point scale for a yes or no response. Confidence rating was significantly higher for the hits to the critical presented words and to the list words than for the false alarms to the critical non-presented words. Experiment 3 further showed that how similar false memory experience was to that of true memory was a function of retention size (number of lists of words retained in memory). In all three experiments, the participants' recognition RTs distinguished false memory from veridical memory, and in Experiments 2 and 3, so did their confidence ratings. Therefore, false memory and veridical memory differ at both the objective and the subjective levels. The results are consistent with a single familiarity dimension model of recognition memory. PMID- 15552360 TI - Conceptual and perceptual information both influence melody identification. AB - Three processes have been identified as central to object identification: top down processing, bottom-up processing, and lateral competition. Six experiments using the perceptual interference paradigm were conducted to assess the relative contributions of these three processes to melody identification. Significant interference was observed only when the target and the distracting information were difficult to distinguish both perceptually and conceptually. Lateral competition-the activation of specific distractor melodies--did not influence the magnitude of interference observed. These results suggest that bottom-up and top down processes contribute more to melody identification than does lateral competition. The data are discussed in terms of the broader literature on object identification and the relationship between identifying melodies, spoken words, and visual objects. PMID- 15552361 TI - Spatial representation by young infants: categorization of spatial relations or sensitivity to a crossing primitive? AB - The spatial representation abilities of 3- to 4-month-old infants were examined in four experiments. Experiments 1 and 2 showed that infants familiarized with a diamond appearing in distinct locations to the left or right of a vertical bar subsequently preferred a stimulus depicting the diamond on the opposite side of the bar over a stimulus depicting the diamond in a novel location on the same side of the bar. Experiment 3 was a replication of Experiment 1, except that the bar was oriented at 45 degrees. In this instance, infants divided their attention between the stimulus depicting the diamond on the opposite side of the bar and the stimulus depicting the diamond in a novel location on the same side of the bar. Experiment 4 demonstrated that the results of Experiment 3 were not a consequence of a failure to process the diagonal bar. When considered with previous reports that infants can represent the categories of above and below (Quinn, 1994), the present results suggest that (1) infants can also represent the categories of left and right, and (2) performance cannot be interpreted as a response to an arbitrary crossing of one object relative to another. Although recent discussions of the relation between language and cognition have pointed to the ways in which spatial language influences spatial cognition (Bowerman & Levinson, 2001), the present findings are consistent with an influence in the opposite direction: Spatial cognition may in some instances shape spatial language. PMID- 15552362 TI - [Analysis of excitatory and inhibitory components of postsynaptic currents recorded in the pyramidal neurons and interneurons of the rat hippocampus]. AB - Postsynaptic currents recorded in the whole-cell configuration with patch-clamp method are actually the sum ofexcitatory (EPSC) and inhibitory (IPSC) components. An approach has been developed allowing the quantitative evaluation of the amplitude and the time course of EPSC and IPSC without treatment of the brain slice with pharmacological inhibitors. The approach is based on the substantial difference in the equilibrium potential values of incoming cationic and anionic currents as the existence of linear regions of corrent-voltage dependence of these currents. The comparison of the results obtained with the classical pharmacological method and with the suggested one demonstrated their coincidence. It allows analysing the postsynaptic currents in sigle neurons without altering the synaptic transmission in the whole brain slice. The contribution of inhibitory currents in the composite synaptic response of intemeurons turned out to be smaller in comparison with pyramidal neurons of CA1 field of the rat hippocampus. PMID- 15552363 TI - [Glutamatergic modulation of vertebrate neuromuscular transmission]. AB - The paper is devoted to the analysis of evidence pointing to presence of glutamatergic modulation of vertebrate neuromuscular transmission. The data on the glutamate's origin and release in the endplate region as well as on the presence of specific glutamate receptors are discussed. The effects of glutamate on different types of acetylcholine secretion in the synapses of amphibians and mammals are described. The question of possible physiological role of glutamatergic modulation of neuromuscular transmission is discussed. PMID- 15552364 TI - [Antiadhesive molecule T-cadherin is an atypical low-density lipoprotein receptor in vascular cells]. AB - Elevated serum LDL level, which results in cholesterol accumulation in vascular wall, is widely accepted as a risk factor in atherosclerosis development. Additionally to metabolic effects, LDL can produce hormone-like effects in a number of cells: activate second messenger systems, regulate gene expression and activate platelets and stimulate cell proliferation. The responses elicited by LDL are rapid, dose-dependent and capable of being saturated, indicating the involvement of specific receptor/binding sites in LDI-stimulated signal transduction. This LDL-binding protein was isolated from human aorta media and identified as T-cadherin. Cadherins are a superfamily of adhesion molecules that mediate Ca2+ -dependent cell-cell adhesion in embryogenesis and in adult organism's solid tissues. Intercellular junctions are formed as a result of interactions between extracellular domains of the neighboring cells' cadherins. Binding of the intercellular domain to the acting cytoskeleton ensures stability of cadherin-mediated adhesive junctions. T-cadherin is a unique member of calcium dependent adherent proteins; in contrast to classical cadherins T-cadherin is anchored to the cell surface membranes via a glycosyl phosphatidyl inositol (GPI) moiety. Subcellular distribution of T-cadherin is restricted to lipid rafts on the cell membranes where it co-localizes with signal-transducing molecules. The function of T-cadherin has not yet been revealed. It was originally cloned from chicken embryo brain where the spatial-temporally restricted pattern of T cadherin suggests its role as a negative guidance cue in tegulating the segmental organization of trunk neural crest migration and motor axon projections. Comparative study of the T-cadherin expression in human organs and tissues revealed that T-cadherin content was maximal in cardiovascular system. Its expression in VSMC depends on the cell phenotype and proliferate activity and increases in atherosclerotic lesion and restenosis. T-cadherin seems to play a key role in the regulation of the vascular cell phenotype, migration and growth. We hypothesize that T-cadherin is an anti-adhesive molecule which participates in intercellular interactions informing cells about their environment and regulating migration and proliferation of cells in vascular wall, while LDL interfere with the normal function of T-cadherin. PMID- 15552365 TI - [The nervous system role in inhibition of hemopoiesis]. AB - The paper presents currently available data on the role of nerve signalisation and inhibitory effect of transforming growth factor B, tumour necrosis factor, interleukine-1 in the regulation of hemopoiesis. PMID- 15552366 TI - [Some problems in measuring the frequency-resolving power of hearing]. AB - In spite of detailed elaboration of masking methods of measuring the frequency selectivity of hearing, such measurements actually are not in use for diagnostics purpose because of their time-consumption and ambiguity of extrapolation of the results to perception of complex sound spectrum patterns. A method of direct measuring of spectrum resolving power using rippled-noise test, is suggested. Results of measurements have shown that the actual ability of hearing to discriminate complex sound spectra is higher than that predicted by acuteness of auditory frequency filters: dependence of acuteness of auditory frequency filters on sound level does not influence the ability to discriminate complex spectra; and the influence on interfering noise on the frequency resolving power can not be explained by a decrease of the spectral contrast by the spread of excitation. PMID- 15552367 TI - [Visual cycle and dark adaptation: a new approach in research]. AB - Visual cycle is the series of reactions that support regeneration of the visual pigmen after its photolysis in retinal rods and cones. Inherited or acquired deficiencies of the visual cycle impair dark adaptation and lead to a series of visual disorders. The paper describes a new approach to study of the visual cycle that uses fast dichroic microspectrophotometer. The method allows studying interconversion of bleaching products in single intact photoreceptors in condition approaching the situation in vivo. Using this approach, we established a complete scheme of transitions between metarhodopsins, retinal and retinol in amphibian rods. It appeared that the decay of metarhodopsins controls both the time course of rod dark adaptation following small bleaches and the production of retinol that is the substrate for rhodopsin regeneration. We also obtained novel data on kinetics of the decay of cone metapigments that was found to be by an order of magnitude faster than in rods. Possible application of the method for further study of the visual cycle in normal and pathological conditions is discussed. PMID- 15552368 TI - [The functional condition of the immune system and the lymphocytes' hemopoiesis modulating properties]. AB - Donor splenocytes and timocytes have an ability to stimulate erythropoesis after massive blood-letting. Changing of functional state of the immune system by means of immune modulators action affects the character and expressiveness of hemopoiesis regulation function of lymphocytes. Splinocytes and thymocytes depress granulocytopoiesis in the recipients' bone marrow after activation by T- and B-lymphocytes. The activation of B-lymphocytes determines the cells' capacity to increase concentration of thrombocytes in the blood. Donor thymocytes can activate erythropoiesis and granulocytopoiesis after macrophages stimulation. PMID- 15552369 TI - [Intracellular recording of the frog dorsal root single fibres' responses mediated by the GABAA and 5-HT-receptors]. AB - The effects of GABA, bicuculline and 5-HT on primary afferents in the isolated spinal cord of the frog Rana ridibunda were studied. Bath application of GABA (1 mM) reduced the primary afferent depolarisation (PAD) in IX segment of the spinal cord evoked by X dorsal root stimulation (57 +/- 8% of initial level, n = 5, p < 0.05). The action potentials (AP) recorded in dorsal root afferents was also suppressed under the GABA action (74 +/- 9%, p < 0.05). Bath application of bicuculline (50 microM) reduced the PAD (21 +/- 7%), n = 6, p < 0.05), meanwhile the AP in dorsal root afferents was resistant against the bicuculline action. Bath application of 5-HT (25 microM) depressed the PAD (34 +/- 7%, n = 7, p < 0.05) and the amplitude of the AP recorded from the single afferent fibre in dorsal column (76 +/- 6%, n = 7, p < 0.05). In contrast to GABA, 5-HT more effectively suppressed the late phase of the PAD evoked by X dorsal root stimulation and caused (76 +/- 6%, n = 7, p < 0.05) an alteration of the AP shape. All effects induced by these drugs were reversible. The mechanisms of GABA and 5-HT modulation of spinal cord afferent income are discussed. PMID- 15552370 TI - [The mechanisms of short-term forms of synaptic plasticity]. AB - In experiments on the frog cutaneous pectoris muscle in cases of different external calcium concentrations, using extracellular recording technique, processes of facilitation and depression of transmitter release during the high frequency stimulation were investigated. On the ground of experiments using intracellular mobile calcium buffers BAPTA-AM and EGTA-AM, it was proposed that at least two (low- and high-affinity) calcium-binding sites underlie the facilitation. Both the facilitation and the depression were accompanied by such transformations of underlied of nerve ending responses as changes of the third phase amplitude. Application of potassium channel blockers allowed us to reveal the significant contribution of changes of duration of the AP repolarisation phase and, accordingly, the changes of magnitude of calcium influx to development of facilitation and depression of transmitter release. It was also revealed that, during the high-frequency rhythmic stimulation, the increase of asynchrony of transmitter release leading to decrease of facilitation and increase of depression occurred. It was concluded that the forms of short-term synaptic plasticity--facilitation and depression, were caused by various presynaptic mechanisms: the increase of concentration of "local" and accumulation of "residual" calcium, the changes of calcium influx, increase of temporal course of secretion, the impairment of equilibrium between the depletion and restoration of mediator supply. Due to some of these processes and specific conditions of synapse functioning, the facilitation of the depression of transmitter release occurred. PMID- 15552371 TI - [Electromechanical heterogeneity of the myocardium]. AB - Herein we discuss modem data showing that ventricle's working myocardium is highly heterogeneous. Significant transmural differences in electrophysiological and biomechanical properties of cardiomyocytes are reviewed. The reviewed evidence of myocardial heterogeneity constitutes the basis for modem assessment of segmental kinetics of different regions in intact heart. We used muscle duplexes as condensed models of a heterogeneous myocardial system. Experimental data, presented here were obtained both in biological duplexes formed by isolated myocardial preparations and in mathematical models of muscle duplexes. We showed that specific functional heterogeneity of cardiomyocytes, related to their excitation sequence, allowed the myocardium to optimise its contractile function and smooth dispersion of repolarisation. PMID- 15552372 TI - Stop it, now. PMID- 15552373 TI - A call to all nurses: get out there and vote! PMID- 15552374 TI - LPNs and nurses' aides also need protection from back injuries. PMID- 15552375 TI - Called to task for using the term, "fifth vital sign". PMID- 15552376 TI - Hands-on help Torsades de pointes. AB - Torsades de pointes, a French term that literally means "twisting of the points," is a lethal form of ventricular tachycardia named for its hallmark feature: polymorphic QRS complexes that appear to twist around the isoelectric line. Without treatment, few patients survive Torsades de pointes (TdP) once it develops. The key to preventing sudden cardiac death lies in early detection of warning signs. PMID- 15552377 TI - Help patients cope with chemo. AB - Chemotherapy is keeping more and more patients alive, but its side effects can be debilitating. Appropriate nursing care and education can help cancer patients overcome the challenges of this vital treatment. PMID- 15552378 TI - Patient information. Managing chemo's side effects. PMID- 15552379 TI - We stop aggression as soon as it starts. AB - It's a sad reality, but all too many of today's nurses find themselves in a workplace where verbal abuse, threats, and physical assaults are common-place. The attacks seem to come from all sides: unruly patients, volatile family members, abrasive doctors, and angry colleagues. PMID- 15552380 TI - Erectile dysfunction. PMID- 15552381 TI - 6 diet myths (and the facts that debunk them). PMID- 15552382 TI - A sideline within nursing? PMID- 15552383 TI - Big mouth! PMID- 15552384 TI - Analysis of the forms, functions and facilitation of social support in one English county: a way for professionals to improve the quality of health care. AB - Voluntary organisations are an integral part of community care, and the available research indicates the value of their social support role. However, surprisingly little is known about the forms and functions of this support, or the links to the formal support provided by the National Health Service (NHS), so hampering quality improvements. Therefore, a small sample of voluntary service organisations in one English county participated in a pilot study. This involved the staff and users of these organisations, and a geographically linked sample of NHS mental health professionals. Interview data indicated that the voluntary sector users and staff held similarly positive views of the appropriately varied forms and functions of the provided social support, and all participants held unusually similar and positive views of their links, although areas for improvement were suggested by both groups (e.g. links to GPs). PMID- 15552385 TI - A new tool for measurement of process-based performance of multispecialty tertiary care hospitals. AB - There is an increasing need of a model for the process-based performance measurement of multispecialty tertiary care hospitals for quality improvement. Analytic hierarchy process (AHP) is utilized in this study to evolve such a model. Each step in the model was derived by group-discussions and brainstorming sessions among experienced clinicians and managers. This tool was applied to two tertiary care teaching hospitals in Barbados and India. The model enabled identification of specific areas where neither hospital performed very well, and helped to suggest recommendations to improve those areas. AHP is recommended as a valuable tool to measure the process-based performance of multispecialty tertiary care hospitals. PMID- 15552386 TI - Nursing work environment and quality of care: differences between units at the same hospital. AB - The literature suggests that improvements in nurses' work environments may improve the quality of patient care. Furthermore, monitoring the work environment through staff surveys may be a feasible method of identifying opportunities for quality improvement. This study aimed to confirm five proposed sub-scales from the Nursing Work Index - Revised (NWI-R) to assess the nursing work environment and the performance of these sub-scales across different units in a hospital. Data were derived from a cross-sectional survey of 243 nurses from 13 units of a 300-bed university-affiliated hospital in Quebec, Canada, during 2001. Using confirmatory factor analysis, the five subscales were confirmed; three of the sub scales had greater ability to discriminate between units. Using hierarchical regression models, "resource adequacy" was the sub-scale most strongly associated with the perceived quality of care at the last shift. The NWI-R sub-scales are potentially useful for comparison of work environments of different nursing units at the same hospital. PMID- 15552387 TI - Quality management in health care: a 20-year journey. AB - In this article, the total quality programme in the Spanish healthcare system (1986-1992) and the subsequent quality improvement steps that have led to definition and implementation of such an integrated framework, seeking a quality management system and patient safety, are discussed. PMID- 15552388 TI - Quality improvement techniques to improve patient satisfaction. AB - This paper describes several approaches for implementing quality improvement initiatives to improve patient satisfaction, which enables health-care organizations to position themselves for success in today's global and increasingly competitive environment. Specifically, measuring the views of patients, improving patient satisfaction through a community-wide effort, and using a Six Sigma program are discussed. Each of these programs can be an effective mechanism for quality improvement. A key component to quality improvement techniques involves collaborative efforts by all health-care professionals and managers as they seek to increase patient satisfaction. PMID- 15552389 TI - Identification of seniors at risk: process evaluation of a screening and referral program for patients aged > or =75 in a community hospital emergency department. AB - Reports on the authors' experience with a patient safety quality improvement program, intended to reduce the incidence and severity of adverse outcomes for emergency department (ED) patients aged > or = 75. The Identification of Seniors at Risk scale was used for screening, and those at high risk were referred for appropriate intervention. The plan-do-study-act improvement cycle was followed, conducting process evaluation to diagnose and correct implementation difficulties. Reports that: implementing an ED screening and referral program is deceptively difficult; process evaluation multidisciplinary working group meetings are an essential improvement tool; screening inclusion criteria had to be adapted to the subject population in order to make efficient use of staff time; the screening questions and process required ongoing assessment, revision, and local adaptation in order to be useful; and high-risk screening in the ED is critical to a hospital system's ability to anticipate clinical problems; the plan do-study-act improvement cycle is a practical and useful tool for improving quality and systems in a real care setting. PMID- 15552390 TI - Completing the circle: from PD to PDSA. AB - Problem-solving teams, involving front-line staff, are widely used to achieve continuous process improvement. Approaches such as "plan-do-study-act" (PDSA) cycles, are now a core element of many health-care improvement initiatives. This paper evaluates the use of PDSA improvement cycles within the UK National Health Service, using emergency care improvement activity as a source of research evidence. It was found that, despite an abundance of information on how to implement this type of change, many senior professionals still misinterpret how this should work. This has implications for how such methodologies are implemented. There is a long way to go in allowing greater employee involvement, moving much further away from the "management committee" style of change. Care has to be taken to ensure that empowered employees are working to consistent and appropriate objectives. It is important that senior personnel develop process understanding alongside the workforce, rather than simply providing distant support. PMID- 15552391 TI - Quality and more quality. PMID- 15552392 TI - Formulating a health quality improvement strategy for a developing country. AB - The purpose of this paper is to present the idea of a national health quality plan as a way of mobilising and organising effort towards quality improvement, and stimulate debate and interes in the West in the subject, showing how quality improvement can be developed and invigorated by engaging in issues arising from quality strategies in developing countries. Considers why developing countries are concerned to improve the quality of health care and the different methods which can be used. Stresses the importance of recognising the starting-point and what can help and hinder this work. Concentrates on improving the quality of health care, rather than the wider and important question of improving health. Concludes that developing countries do have the exerptise and that extra resources are needed, mostly for training and supervision. Opines that if implementation is not managed then the investment will be wasted. PMID- 15552393 TI - Measuring service quality provision within an eating disorders context. AB - Traditionally, research concerning quality measurement and improvement has centred on manufacturing and industrial sectors. In the last decade, however, the focus has turned to the measurement of quality in services and more recently to healthcare. In today's progressively competitive health-care environment hospitals increasingly realise the need service quality as a means to improve their competitive position. The aim of this study is to assess the service quality provision of a leading UK-based eating disorder service provider, achieved through the development and application of the SERVQUAL research instrument for measuring service quality. Following thematic analysis of service user feedback, six dimensions of service quality were identified and mapped on to the P-C-P attributes model. The approach adopted for this case study research may usefully inform service assessment programmes for similarly positioned health care service providers. PMID- 15552394 TI - A critique of emerging European legislation in the pharmaceutical industry: a clinical trials analysis. AB - The objective of this paper is to critique emerging legislation in the pharmaceutical industry, focusing on the clinical trials sector. Possible changes are identified and discussed inrelation to their impact on phase I clinical trials conducted in the UK. It is concluded that smaller contract research organisations, which have benefited in the past from European Union legislative variation, may have resource problems in trying to cope with the changing business environment created through legislative harmonization. These SMEs must use this opportunity to seek clinical trials research partnerships in a new harmonized EU market. PMID- 15552395 TI - Relationship between malpractice claims and medical care quality. AB - The purpose of this paper was to examine the relationship between medical malpractice claims and medical care quality in Taiwan. The Delphi technique with an expert panel was used to determine the relationship between malpractice and medical quality. A total of 371 medical malpractice claims were analyzed. Main measures included the rate and strength of malpractice cases associated with quality and the identification of the quality factors influencing the occurrence of malpractice. Results showed that malpractice claims were associated with internal medicine cases, surgery cases, pediatric cases, obstetricc and gynecological cases, physicians' professional competence, nonacceptable outcomes, complications, and poor communication. Concludes that medical malpractice cases could be avoided by increasing physicians' professional knowledge, practical skills, and communication. PMID- 15552396 TI - Developing service quality in mental healtlh services. AB - Through a literature review, this paper demonstrates that there is a lack of knowledge about quality management within the mental health services. Ideas about quality from the wider services sector are then discussed, and a case study of a mental health scenario is provided. It is argued that health service generally, and mental health services in particular, have much to gain from becoming more closely aligned with the wider field of knowledge of quality management. Concludes that the wider techniques of managing service quality may prove useful, particularly in mental health services, due to the nature of such services and their inherent variability. PMID- 15552397 TI - The multiple roles of the mitochondrion of the malarial parasite. AB - Mitochondria of the malaria parasite Plasmodium falciparum are morphologically different between the asexual and sexual blood stages (gametocytes). In this paper recent findings of mitochondrial heterogeneity are reviewed based on their ultrastructural characteristics, metabolic activities and the differential expression of their genes in these 2 blood stages of the parasite. The existence of NADH dehydrogenase (complex I), succinate dehydrogenase (complex II), cytochrome c reductase (complex III) and cytochrome c oxidase (complex IV) suggests that the biochemically active electron transport system operates in this parasite. There is also an alternative electron transport branch pathway, including an anaerobic function of complex II. One of the functional roles of the mitochondrion in the parasite is the coordination of pyrimidine biosynthesis, the electron transport system and oxygen utilization via dihydroorotate dehydrogenase and coenzyme Q. Complete sets of genes encoding enzymes of the tricarboxylic acid cycle and the ATP synthase complex are predicted from P. falciparum genomics information. Other metabolic roles of this organelle include membrane potential maintenance, haem and coenzyme Q biosynthesis, and oxidative phosphorylation. Furthermore, the mitochondrion may be a chemotherapeutic target for antimalarial drug development. The antimalarial drug atovaquone targets the mitochondrion. PMID- 15552398 TI - Activity of benzothiazoles and chemical derivatives on Plasmodium falciparum. AB - Malaria is a major health concern particularly in Africa which has about 90% of the worldwide annual clinical cases. The increasing number of drug-resistant Plasmodium falciparum justifies the search for new drugs in this field. Antimalarial activity of 2-substituted 6-nitro- and 6-amino-benzothiazoles and their anthranilic acids has been tested. An in vitro study has been performed on W2 and 3D7 strains of P. falciparum and on clinical isolates from malaria infected patients. Toxicity has been assessed on THP1 human monocytic cells. For the most active drug candidates, the in vitro study was followed by in vivo assays on P. berghei-infected mice and by in vitro assays in order to determine the stage-dependency and the mechanism of action. Of 39 derivatives tested in vitro, 2 had specific antimalarial properties. Each compound was active on all stages of the parasite, but one was markedly active on mature schizonts, while the other was more active on young schizont forms. Both drugs were also active on mitochondrial membrane potential. In vivo data confirmed efficiency with a sustained decrease of parasitaemia. Products A12 and C7 may be considered as potential antimalarial worthy of further chemical and biological research. PMID- 15552399 TI - Trypanosomatid biodiversity in Costa Rica: genotyping of parasites from Heteroptera using the spliced leader RNA gene. AB - The biodiversity of insect trypanosomes is largely unknown, resulting in significant gaps in the understanding of pathogen evolution. A culture independent preliminary survey of trypanosomatid fauna was conducted for the parasites of Heteroptera (Hemiptera) from several localities in Costa Rica. Trypanosomatid infections were detected by light microscopy of smeared gut contents. Out of 257 insects representing 6 families, infections were found in 62 cases; cultures were obtained for 29 new isolates. Gut material from infected hosts was preserved in the field using an SDS-EDTA buffer solution for subsequent DNA extraction in the laboratory. PCR amplification of the trypanosomatid specific spliced leader (SL) RNA gene repeats was successful for 60 field samples. Eighteen distinct SL RNA typing units were identified in a set of 28 samples analysed in detail. Cluster analysis indicated that these typing units were unique and thus could represent new species and, in some cases, new genera. This study reveals only a minor fraction of the trypanosomatid biodiversity, which is anticipated to be high. PMID- 15552400 TI - Phylogeny, taxonomy and grouping of Trypanosoma rangeli isolates from man, triatomines and sylvatic mammals from widespread geographical origin based on SSU and ITS ribosomal sequences. AB - Phylogenetic relationships among Trypanosoma rangeli isolates from man, wild mammals and triatomine bugs from widespread geographical origin were inferred by comparison of the small subunit of ribosomal gene sequences. The phylogenetic trees indicated that the subgenus Herpetosoma is polyphyletic and strongly supported division of this group into two monophyletic lineages, one made up of T. rangeli, T. rangeli-like and allied species and other consisting of T. lewisi and related taxa. Based on phylogenetic analysis, morphology, behaviour in vertebrate and invertebrate hosts and epidemiology we propose: a) the validation of Herpetosoma as a taxon comprised only for species of group lewisi and the maintenance of T. lewisi as the type-species of this subgenus; b) the classification of T. rangeli, T. rangeli-like and allied species into a 'T. rangeli-clade' more closely related to Schizotrypanum than to T. lewisi or T. brucei. The phylogenetic tree disclosed at least 4 groups within the clade T. rangeli, all confirmed by polymorphism of the internal transcribed spacer, thus conferring for the first time phylogenetic support to groups of T. rangeli and corroborating the high complexity of this taxon. Grouping was independent of their mammalian host-species and geographical origin, indicating that other factors are determining this segregation. PMID- 15552401 TI - Immunization with cytoplasmic repetitive antigen and flagellar repetitive antigen of Trypanosoma cruzi stimulates a cellular immune response in mice. AB - In previous studies, we demonstrated that CRA and FRA recombinant proteins, used for diagnosis of Chagas' disease, elicited a humoral immune response in susceptible and resistant mice. To understand better the immune response to these proteins, we have evaluated, the cellular immune response in CRA- and in FRA immunized BALB/c and C57BL/6 mice. A specific cellular lymphoproliferative response was observed in both strains of mice. Spleen cell cultures mainly from CRA-immunized C57BL/6 and FRA-immunized BALB/c mice produced high levels of IFN y, indicating the induction of a Type 1 immune response. Regarding the T cell subsets, CD4+ T cells were the major source of IFN-y in CRA- and FRA-immunized mice. These results suggest that CRA and FRA are important immunogens in inducing a Type 1 immune response and that they may be considered as potential vaccine antigens. PMID- 15552402 TI - Molecular ecology of Schistosoma mansoni transmission inferred from the genetic composition of larval and adult infrapopulations within intermediate and definitive hosts. AB - We investigated the genotypic composition of the digenetic parasite Schistosoma mansoni for its adult stages within the definitive host (the wild rat, Rattus rattus) and for the larval stages within the intermediate host (the snail, Biomphalaria glabrata) both collected at the same transmission site. Our analyses are based upon the recognition and distribution of 200 different multilocus genotypes generated by RAPD markers. While intramolluscan larval infrapopulations are characterized by a low infection rate (0.6 % on average) and low intra-host genetic diversity (1.1 genotype on average per infected snail), adult infrapopulations within rats showed a high infection rate (94%) and a substantial intra-host genetic diversity (34 genotypes on average) linked to high intensities (160 worms per host on average). A single definitive host bearing 105 different genotypes harboured 52 % of the total genetic diversity detected within the whole parasite population. Analysis of the genetic data allowed the identification of various ecological, behavioural and immunological factors which are likely to enhance transmission of multiple parasite genotypes towards the vertebrate hosts. From the distribution of repeated identical multilocus genotypes within the parasite population and among the hosts, we have inferred different parameters of the cercarial transmission efficiency as well as patterns and processes by which vertebrate hosts acquire infection in the field. PMID- 15552403 TI - Stage-specific alternative splicing of the heat-shock transcription factor during the life-cycle of Schistosoma mansoni. AB - Stage-specific alternative splicing of the heat-shock transcription factor of Schistosoma mansoni (SmHSF) generates isoforms with structural diversity that may modulate the activity of SmHSF at different life-stages, and thus may regulate the expression of different genes at different developmental stages. RT-PCR, cloning and DNA-sequence analyses showed stage-specific alternative splicing inside the DNA-binding domain (DBD) involving introns I1 and I2, and beyond the DBD involving introns I4a and I7. Retention of introns I2 and I4 would inactivate SmHSF since they contain termination codons. Retention of intron I1 would add 11 amino acids inside the DBD and may change the DNA-binding specificity of SmHSF; intron I7 would add 13 amino acids to the effector region of HSF. Retention of introns was more pronounced in cercariae (larval stage living in water) than in adult worms (parasitic form in mammals). The isoforms were expressed in bacteria, but functional evaluation was not feasible, because only the isoform lacking introns was soluble while isoforms with introns were insoluble. However, stage specific alternative splicing that changed HSF function in vivo was evidenced in intact cercariae. The cercarial SmHSF mRNA was enriched with introns I2 and I4a that contain termination codons. Therefore, translation of the SmHSF mRNA was impaired, and the SmHSF protein was undetectable. Consequently, the HSP70 gene could not be transcribed, and the HSP70 mRNA was missing. Alternative splicing was observed for short DNA segments (33-45 bp) bound by splice signals, located in the coding region. These are not bona fida exons since they are not flanked by introns. Yet, they are not regular introns since they are often found in mature mRNA. Alternative splicing of these DNA segments caused structural diversity that could modulate the function of the gene product. PMID- 15552404 TI - The effect of colostrum on pigs pre-natally or post-natally exposed to Schistosoma japonicum. AB - Pre-natal infection of Schistosoma japonicum in pigs may prove to be a useful model in shedding light on human pre-natal schistosomiasis. This study describes the effects of immune colostrum on worm burdens, tissue egg counts, liver pathology and crude worm or egg antigen-specific IgG and IgA responses, in groups of pigs pre-natally, pre-natally + post-natally or post-natally exposed to S. japonicum. Results suggest that pre-natal exposure and immune colostrum did not affect the establishment of a post-natal challenge infection. However, immune colostrum seemed to increase the levels of septal fibrosis in pre-natally exposed pigs. These findings indicate that further investigations will prove valuable, elucidating the influence of the parasitological and immunological status of the sow, on pre-natally exposed pigs, and on the ability of these pigs to develop resistance against S. japonicum later in life. PMID- 15552405 TI - Meta-analysis of age-prevalence patterns in lymphatic filariasis: no decline in microfilaraemia prevalence in older age groups as predicted by models with acquired immunity. AB - The role of acquired immunity in lymphatic filariasis is uncertain. Assuming that immunity against new infections develops gradually with accumulated experience of infection, models predict a decline in prevalence after teenage or early adulthood. A strong indication for acquired immunity was found in longitudinal data from Pondicherry, India, where Mf prevalence was highest around the age of 20 and declined thereafter. We reviewed published studies from India and Subsaharan Africa to investigate whether their age-prevalence patterns support the models with acquired immunity. By comparing prevalence levels in 2 adult age groups we tested whether prevalence declined at older age. For India, comparison of age groups 20-39 and 40+ revealed a significant decline in only 6 out of 53 sites, whereas a significant increase occurred more often (10 sites). Comparison of older age groups provided no indication that a decline would start at a later age. Results from Africa were even more striking, with many more significant increases than declines, irrespective of the age groups compared. The occurrence of a decline was not related to the overall Mf prevalence and seems to be a chance finding. We conclude that there is no evidence of a general age-prevalence pattern that would correspond to the acquired immunity models. The Pondicherry study is an exceptional situation that may have guided us in the wrong direction. PMID- 15552406 TI - Structure of the microfilarial reservoir of Loa loa in the human host and its implications for monitoring the progr,ammes of Community-Directed Treatment with Ivermectin carried out in Africa. AB - This paper describes the structure of the microfilarial reservoir of Loa loa in an endemic population of central Cameroon. The possible effects of age and sex on the prevalence and intensity of microfilaraemia have been explored. Logistic analysis showed that the prevalence of microfilaraemia increased significantly with age, reaching 60 % in the oldest males. This result suggests that the figure commonly reported, according to which only one third of the infected individuals were microfilaraemic, should be reconsidered; in addition, as part of surveys of loiasis, crude microfilaraemia prevalence values should be replaced by adjusted ones. The intensity of infection did not show any age-specific change. As a result, even if the oldest members of the male population are clearly the most at risk of developing post-ivermectin serious adverse reactions, especially Loa encephalopathy, the other members of the population are not risk-free. Therefore, in those areas where the African Programme for Onchocerciasis Control is undertaking regular mass distributions of ivermectin for onchocerciasis control, and where loiasis is co-endemic, no subpopulation should be excluded from surveillance and monitoring during community directed treatments with ivermectin. PMID- 15552407 TI - Biology of Thelazia callipaeda (Spirurida, Thelaziidae) eyeworms in naturally infected definitive hosts. AB - Thelazia callipaeda (Spirurida, Thelaziidae) eyeworm causes ocular infection in carnivores and humans in the Far East; this infection has been recently reported also in Europe--northern and southern Italy--in dogs, cats and foxes. The natural vector/s of T. callipaeda is/are unknown and the development of the nematode in its definitive hosts is limited to an experimental trial on dogs. To contribute new insights into the development of T. callipaeda in the definitive host in field conditions, eyeworms were collected from naturally infected dogs from an area with a high prevalence of infection (up to 60.14%) in the Basilicata region of southern Italy, from January 2002 to December 2003. Conjunctival secretions were also collected and examined for the presence of immature stages. The presence of blastomerized eggs throughout the period--except for the months from May to November--indicates a seasonality in the reproductive activity of T. callipaeda, coinciding with the presence/absence of the vector. In fact, 1st stage larvae were found in the lachrymal secretions of dogs in summer (June--July 2002 and 2003), ready to be ingested by flies feeding about the eyes. The evidence of 4th-stage larvae in March 2002 and April, July and October 2003 may be accounted for by the presence of flies that act as intermediate hosts of T. callipaeda from early spring to early autumn. The presence of immature stages in October indicates an overlapping generation of nematodes and a 2nd cycle of vector infection. This basic knowledge of the development of T. callipaeda will hopefully help future epidemiological studies to identify the intermediate hosts and define the likely risk for vectors in field conditions. PMID- 15552408 TI - Stability of Corynosoma populations with fluctuating population densities of the seal definitive host. AB - In theory there should be a strong coupling between host and parasite population sizes. Here, we investigated population size and structure in 3 species of acanthocephalans, Corynosoma semerne, C. strumosum and C. magdaleni, in ringed seals (Phoca hispida) from the Bothnian Bay over a period of more than 20 years. During this period, seal numbers first decreased markedly and then increased steadily; at the same time, a paratenic fish host particularly important for C. strumosum has gradually disappeared from the bay due to decreasing salinity. We found no evidence that the mean abundance of any of the 3 acanthocephalan species changed significantly over time, nor was there any relationship between parasite abundance at any point in time and seal numbers in the corresponding year. Based on the proportion of sexually mature female worms per infrapopulation, and on relationships between the sex ratio of worms and infrapopulation size, both C. magdaleni and C. semerme appear to be doing well, independently of the population size of their seal definitive hosts. In contrast, perhaps because of the loss of its main paratenic host, C. strumosum appears more at risk in the Bothnian Bay. Our results show that in complex natural systems, there are not necessarily simple, direct links between definitive host population size or density, and parasite population dynamics. PMID- 15552409 TI - Dynamics of chemical mimicry in the social parasite wasp Polistes semenowi (Hymenoptera: Vespidae). AB - Chemical cues are so important in the recognition mechanism of social insects that most social parasites (which rely on hosts to rear their brood) have been documented as overcoming the mechanism by which colony residents recognize non nestmates, by mimicking the odour of the usurped colony. We simulated in the laboratory the process by which the obligate social parasite, Polistes semenowi, invades nests of the host species, Polistes dominulus, in the field and analysed the epicuticular lipid layer before and after host nest usurpation. The experiment documents that P. semenowi social parasites have an epicuticular hydrocarbon pattern which is very similar to that of their host but, after entering host colonies, parasites mimic the odour of the colonies they invade, to the point that they perfectly match the hydrocarbon profile peculiar to the colony they entered. However, both before and after host nest invasion, parasites show a tendency to possess diluted recognition cues with respect to their hosts. PMID- 15552410 TI - Temperature distribution in atherosclerotic coronary arteries: influence of plaque geometry and flow (a numerical study). AB - Intravascular coronary thermography is a method that may detect vulnerable, atherosclerotic plaques and is currently evaluated in a clinical setting. Active macrophages or enzymatic heat releasing processes in vulnerable plaques may act as heat sources. To better understand the parameters of influence on thermographic measurements, numerical simulations have been performed on a model of a coronary artery segment containing a heat source. Heat source parameters and flow were varied to study their influence on temperatures at the lumen wall. Maximal temperature differences at the lumen wall increased when the source volume increased and they differ with the source geometry. The simulations showed that blood flow acts as a coolant to the lumen wall. Blood flow decreased maximal temperatures depending on the source geometry, source volume and the maximal flow velocity. Influence of flow was highest for circumferentially extended sources, up to a factor 3.7, and lowest for longitudinally extended sources, down to a factor 1.9. When cap thickness increased, maximal temperatures decreased and the influence of flow increased. This study shows that correct interpretation of intravascular thermographic measurements requires data on the flow and on the morphologic characteristics of the atherosclerotic plaque. PMID- 15552411 TI - The relationship between temporal variation of hypoxia, polarographic measurements and predictions of tumour response to radiation. AB - The polarographic oxygen sensor is one of the most used devices for in vivo measurements of oxygen and many other measurement techniques for measuring tumour hypoxia are correlated with electrode measurements. Little is known however about the relationship between electrode measurements and the real tissue oxygenation. This paper investigates the influence of the temporal change of the hypoxic pattern on the electrode measurements and the tumour response. Electrode measurements and tumour response were simulated using a computer program that allows both the calculation of the tissue oxygenation with respect to the two types of hypoxia that might arise in tumours and the virtual insertion of the electrode into the tissue. It was therefore possible to control the amount of each type of hypoxia in order to investigate their influence on the measurement results. Tissues with several vascular architectures ranging from well oxygenated to poorly oxygenated were taken into consideration as might be seen in practice. The influence of the electrode measurements on the treatment outcome was estimated by calculating the tumour control probability for the tumours characterized either by the real or by the measured tumour oxygenation. We have simulated electrode oxygen measurements in different types of tissues, covering a wide range of tumour oxygenations. The results of the simulations showed that the measured distribution depends on the details of the vascular network and not on the type of hypoxia. We have also simulated the effects of the temporal change of the acute hypoxic pattern due to the opening and the closure of different blood vessels during a full fractionated treatment. The results of this simulation suggested that the temporal variation of the hypoxic pattern does not lead to significantly different results for the electrode measurements or the predicted tumour control probabilities. In conclusion, it was found that the averaging effect of the electrode leads to a systematic deviation between the actual oxygen distribution and the measured distribution. However, as the electrode reflects the general trends of the tissue oxygenation it has the potential of being used for the general characterization of tumour hypoxia even if the actual type of hypoxia measured by the electrode cannot be determined. Indeed, the change in time of the acute hypoxic region does not compensate for the lack of oxygenation at a specific moment and therefore does not influence the polarographic oxygen measurements. PMID- 15552412 TI - Comparison of in vitro and in vivo alpha/beta ratios for prostate cancer. AB - Parallel in vitro and in vivo studies provide insight into the relationship between clinical response and intrinsic cellular radiosensitivity and may aid in the development of predictive assays. Compilations of radiosensitivity parameters from in vitro experiments can also be used to examine the potential effectiveness of alternative or new treatment plan designs until enough clinical data become available to directly estimate the requisite radiosensitivity parameters. In this work, survival data for six prostate cancer cell lines (ten datasets total) have been extracted from the literature and re-analysed using the linear-quadratic (LQ) survival model. The paired bootstrap technique for regression is used to compute 95% confidence intervals for the estimated radiosensitivity parameters. LQ radiosensitivity parameters derived from the in vitro data are then compared to radiosensitivity parameters derived from clinical data for prostate cancer. Estimates of alpha range from 0.09 to 0.35 Gy(-1) (all cell lines), and the alpha/beta ratio ranges from 1.09 to 6.29 Gy (all cell lines). Point estimates of the repair half-time (PPC-1, TSU-Pr1, PC-3 and DU-145 cell lines) range from 5.7 to 8.9 h (95% confidence interval from 0.26 h to 10.7 h). Differences in the radiosensitivity parameters determined from the data reported by different laboratories are as large as or larger than the differences in radiosensitivity parameters observed among the various prostate cell lines. The reported studies demonstrate that even seemingly small corrections for dose rate effects, such as those expected in high dose rate (HDR) experiments, can sometimes have a significant impact on estimates of alpha and alpha/beta. By neglecting dose rate effects in the analysis of HDR experiments, estimates of the alpha/beta, ratio may be too high by factors as large as 1.3 to 6.2. The half-time for repair derived from the in vitro experiments appears significantly larger (slower repair rate) than estimates derived from the clinical data. However, the prostate radiosensitivity parameters alpha and alpha/beta may be approximately the same in vitro and in vivo. Most of the in vitro data are consistent with an alpha/beta ratio for prostate cancer less than 3 or 4 Gy. PMID- 15552413 TI - Measurements of output factors with different detector types and Monte Carlo calculations of stopping-power ratios for degraded electron beams. AB - The aim of the present study was to investigate three different detector types (a parallel-plate ionization chamber, a p-type silicon diode and a diamond detector) with regard to output factor measurements in degraded electron beams, such as those encountered in small-electron-field radiotherapy and intraoperative radiation therapy (IORT). The Monte Carlo method was used to calculate mass collision stopping-power ratios between water and the different detector materials for these complex electron beams (nominal energies of 6, 12 and 20 MeV). The diamond detector was shown to exhibit excellent properties for output factor measurements in degraded beams and was therefore used as a reference. The diode detector was found to be well suited for practical measurements of output factors, although the water-to-silicon stopping-power ratio was shown to vary slightly with treatment set-up and irradiation depth (especially for lower electron energies). Application of ionization-chamber-based dosimetry, according to international dosimetry protocols, will introduce uncertainties smaller than 0.3% into the output factor determination for conventional IORT beams if the variation of the water-to-air stopping-power ratio is not taken into account. The IORT system at our department includes a 0.3 cm thin plastic scatterer inside the therapeutic beam, which furthermore increases the energy degradation of the electrons. By ignoring the change in the water-to-air stopping-power ratio due to this scatterer, the output factor could be underestimated by up to 1.3%. This was verified by the measurements. In small-electron-beam dosimetry, the water-to-air stopping-power ratio variation with field size could mostly be ignored. For fields with flat lateral dose profiles (>3 x 3 cm2), output factors determined with the ionization chamber were found to be in close agreement with the results of the diamond detector. For smaller field sizes the lateral extension of the ionization chamber hampers its use. We therefore recommend that the readily available silicon diode detector should be used for output factor measurements in complex electron fields. PMID- 15552414 TI - On the relation between the spatial dose integrity and the temporal instability of polymer gel dosimeters. AB - When irradiating a polymer gel dosimeter to relatively high doses, edge enhancing effects (overshoots) may be noticed near dose gradients, resulting in a loss of spatial dose integrity. These overshoots are believed to be a consequence of monomers diffusing into the high-dose region, where they react with long-living macroradicals. Macroradicals may also be responsible for the temporal chemical instability of post-irradiation polymerization that occurs in the polymer gel dosimeter. In this study, a mathematical model is proposed that simulates the edge enhancing effect. The model is based on the hypothesis that the macroradicals are responsible for both the temporal instability and loss of spatial dose integrity. All input parameters for the model are obtained from independent experiments. The edge enhancing effect is studied both experimentally and theoretically for polymer gel dosimeters with various gelatin concentrations. The change in the edge enhancement is also investigated over post-irradiation time. Comparisons between polymer gel measurements and simulations confirm the hypothesis that there is a strong relation between the spatial and temporal instabilities. PMID- 15552415 TI - Experimental kerma coefficients for carbon deduced from microscopic cross sections at 96 MeV incident neutron energy. AB - The double-differential cross sections for (n, px), (n, dx), (n, tx), (n, 3Hex) and (n, alphax) reactions in carbon have been measured at 96 MeV incident neutron energy. The various charged particles (inclusive spectra) were identified using deltaE-E techniques. From the experimental data, energy- and angle-differential as well as production cross sections were determined, and subsequently the partial and total kerma coefficients. The deduced partial and total kerma coefficients were compared to previous experimental results and theoretical calculations. The findings indicate that the deduced kerma coefficients for the hydrogen isotopes are in good agreement with those deduced from a previous measurement, and that the kerma coefficient values, in particular of the hydrogen isotopes, are systematically higher than values obtained from recent model calculations, which consequently resulted in a total kerma coefficient which is up to 30% higher than predicted by the calculations. PMID- 15552417 TI - Pragmatic fully 3D image reconstruction for the MiCES mouse imaging PET scanner. AB - We present a pragmatic approach to image reconstruction for data from the micro crystal elements system (MiCES) fully 3D mouse imaging positron emission tomography (PET) scanner under construction at the University of Washington. Our approach is modelled on fully 3D image reconstruction used in clinical PET scanners, which is based on Fourier rebinning (FORE) followed by 2D iterative image reconstruction using ordered-subsets expectation-maximization (OSEM). The use of iterative methods allows modelling of physical effects (e.g., statistical noise, detector blurring, attenuation, etc), while FORE accelerates the reconstruction process by reducing the fully 3D data to a stacked set of independent 2D sinograms. Previous investigations have indicated that non stationary detector point-spread response effects, which are typically ignored for clinical imaging, significantly impact image quality for the MiCES scanner geometry. To model the effect of non-stationary detector blurring (DB) in the FORE+OSEM(DB) algorithm, we have added a factorized system matrix to the ASPIRE reconstruction library. Initial results indicate that the proposed approach produces an improvement in resolution without an undue increase in noise and without a significant increase in the computational burden. The impact on task performance, however, remains to be evaluated. PMID- 15552416 TI - GATE: a simulation toolkit for PET and SPECT. AB - Monte Carlo simulation is an essential tool in emission tomography that can assist in the design of new medical imaging devices, the optimization of acquisition protocols and the development or assessment of image reconstruction algorithms and correction techniques. GATE, the Geant4 Application for Tomographic Emission, encapsulates the Geant4 libraries to achieve a modular, versatile, scripted simulation toolkit adapted to the field of nuclear medicine. In particular, GATE allows the description of time-dependent phenomena such as source or detector movement, and source decay kinetics. This feature makes it possible to simulate time curves under realistic acquisition conditions and to test dynamic reconstruction algorithms. This paper gives a detailed description of the design and development of GATE by the OpenGATE collaboration, whose continuing objective is to improve, document and validate GATE by simulating commercially available imaging systems for PET and SPECT. Large effort is also invested in the ability and the flexibility to model novel detection systems or systems still under design. A public release of GATE licensed under the GNU Lesser General Public License can be downloaded at http:/www-lphe.epfl.ch/GATE/. Two benchmarks developed for PET and SPECT to test the installation of GATE and to serve as a tutorial for the users are presented. Extensive validation of the GATE simulation platform has been started, comparing simulations and measurements on commercially available acquisition systems. References to those results are listed. The future prospects towards the gridification of GATE and its extension to other domains such as dosimetry are also discussed. PMID- 15552418 TI - Design and simulation of a high-resolution stationary SPECT system for small animals. AB - Exciting new SPECT systems can be created by combining pinhole imaging with compact high-resolution gamma cameras. These new systems are able to solve the problem of the limited sensitivity-resolution trade-off that hampers contemporary small animal SPECT. The design presented here (U-SPECT-III) uses a set of detectors placed in a polygonal configuration and a cylindrical collimator that contains 135 pinholes arranged in nine rings. Each ring contains 15 gold pinhole apertures that focus on the centre of the cylinder. A non-overlapping projection is acquired via each pinhole. Consequently, when a mouse brain is placed in the central field-of-view, each voxel in the cerebrum can be observed via 130 to 135 different pinholes simultaneously. A method for high-resolution scintillation detection is described that eliminates the depth-of-interaction problem encountered with pinhole cameras, and is expected to provide intrinsic detector resolutions better than 150 microm. By means of simulations U-SPECT-III is compared to a simulated dual pinhole SPECT (DP-SPECT) system with a pixelated array consisting of 2.0 x 2.0 mm NaI crystals. Analytic calculations indicate that the proposed U-SPECT-III system yields an almost four times higher linear and about sixty times higher volumetric system resolution than DP-SPECT, when the systems are compared at matching system sensitivity. In addition, it should be possible to achieve a 15 up to 30 times higher sensitivity with U-SPECT-III when the systems are compared at equal resolution. Simulated images of a digital mouse brain phantom show much more detail with U-SPECT-III than with DP-SPECT. In a resolution phantom, 0.3 mm diameter cold rods are clearly visible with U-SPECT III, whereas with DP-SPECT the smallest visible rods are about 0.6-0.8 mm. Furthermore, with U-SPECT-III, the image deformations outside the central plane of reconstruction that hamper conventional pinhole SPECT are strongly suppressed. Simulation results indicate that future pinhole SPECT systems are likely to bring about significant improvements in radio-molecular imaging of small animals. PMID- 15552419 TI - Image quality assessment of LaBr3-based whole-body 3D PET scanners: a Monte Carlo evaluation. AB - The main thrust for this work is the investigation and design of a whole-body PET scanner based on new lanthanum bromide scintillators. We use Monte Carlo simulations to generate data for a 3D PET scanner based on LaBr3 detectors, and to assess the count-rate capability and the reconstructed image quality of phantoms with hot and cold spheres using contrast and noise parameters. Previously we have shown that LaBr3 has very high light output, excellent energy resolution and fast timing properties which can lead to the design of a time-of flight (TOF) whole-body PET camera. The data presented here illustrate the performance of LaBr3 without the additional benefit of TOF information, although our intention is to develop a scanner with TOF measurement capability. The only drawbacks of LaBr3 are the lower stopping power and photo-fraction which affect both sensitivity and spatial resolution. However, in 3D PET imaging where energy resolution is very important for reducing scattered coincidences in the reconstructed image, the image quality attained in a non-TOF LaBr3 scanner can potentially equal or surpass that achieved with other high sensitivity scanners. Our results show that there is a gain in NEC arising from the reduced scatter and random fractions in a LaBr3 scanner. The reconstructed image resolution is slightly worse than a high-Z scintillator, but at increased count-rates, reduced pulse pileup leads to an image resolution similar to that of LSO. Image quality simulations predict reduced contrast for small hot spheres compared to an LSO scanner, but improved noise characteristics at similar clinical activity levels. PMID- 15552420 TI - A dose calculation method including scatter for IMRT optimization. AB - IMRT optimization is an iterative process in which many dose calculations may be necessary. A fast and accurate dose calculation engine is essential for the optimization program to converge to a reliable solution in a reasonable time. Previously we developed a fast dose calculation method for IMRT optimization based on the table lookup (TLP). The TLP kernel is constructed from the dose distributions computed from an accurate superposition/convolution (SC) algorithm at the beginning. Dose calculations at subsequent iterations are performed by looking up the kernel in memory, therefore are very fast. However, it is assumed by the TLP that the beamlet only deposits doses to the voxels on its path. In this paper, we propose an improved table lookup based dose calculation method that includes the scatter contribution (TLPS). The intensity map for each beam is decomposed into an even and an odd checkbox-type pattern for the construction of the kernel for primary and scatter components. The performance of this algorithm is compared with the methods of SC only and TLP without scatter on a few clinical cases and the advantages of speed and accuracy are demonstrated. This method is especially useful for those optimizations that utilize the IMRT optimization as an inner loop, such as the beam angle optimization. PMID- 15552421 TI - DOSSCORE: an accelerated DOSXYZnrc code with an efficient stepping algorithm and scoring grid. AB - DOSSCORE is an accelerated version of DOSXYZnrc that allows photons to cross voxel boundaries of the same medium and utilizes a separate scoring grid superimposed on the geometrical grid. Two different stepping algorithms, the hownear method and the scaling method are implemented in DOSSCORE. The hownear method allows particles to travel larger distances in homogeneous regions where there is no interest in the dose deposition of these particles, whilst the scaling method utilizes a stepping algorithm in which particles are only slowed down by the boundaries of the geometrical voxels and not by the boundaries of the scoring voxels. For CT-based phantoms, only photon ray tracing is applied, which results in a rather modest speed gain of factor 1.2 compared to DOSXYZnrc. The hownear method and scaling method do not increase the speed for CT-based phantoms, but only for homogeneous phantoms and phantoms with a limited number of small heterogeneities. In cases where a small number of scoring voxels are needed, the hownear method performs better than the scaling method, whilst the opposite is true for cases when many scoring voxels are needed. The photon transport is accelerated by almost a factor of 2 for all phantoms (homogeneous, heterogeneous with much homogeneity and CT-based phantoms) compared to DOSXYZnrc. For a small number of scoring voxels, the hownear method is up to a factor of 2.6 and 1.9 faster than DOSXYZnrc for homogeneous and heterogeneous phantoms in the case of photon beams. For an electron beam, a speed gain of factor 2.4 is obtained. For a full scoring grid like the one used in DOSXYZnrc, the scaling method is up to a factor of 2.2 and 1.7 faster than DOSXYZnrc for homogeneous and heterogeneous phantoms in the case of photon beams. For an electron beam, a speed gain of factor 2 is obtained. A speed increase without biasing the results is very relevant. The use of two separate grids, the more efficient stepping algorithms and the accelerated photon transport can be applied to every EGS-based or other Monte Carlo code. PMID- 15552422 TI - Development of an inorganic scintillating mixture for proton beam verification dosimetry. AB - The availability at the Paul Scherrer Institute (PSI) of a spot-scanning technique with an isocentric beam delivery system (gantry) allows the realization of intensity-modulated proton therapy (IMPT). The development of 3D dosimetry is an important tool for the verification of IMPT therapy plans based on inhomogeneous 3D conformal dose distributions. For that purpose new dosimeters are being developed. The concept is to use a system of many millimetre sized scintillating volumes distributed in a polyethylene block, which are read on a CCD camera over a bundle of optical fibres and which can be irradiated from any direction orthogonal to the fibre axis. The purpose of this work is to investigate the composition of such small sensitive volumes. A mixture of inorganic phosphors and optical cement allows an optimal coupling between the scintillating volume and the optical fibre. Five different inorganic phosphors, available as powder, have been examined by considering their response along the Bragg curve. In particular, two phosphors have shown interesting behaviours: Gd2O2S:Tb and (Zn, Cd)S:Ag. Both phosphors have a high emission efficiency but contrasting behaviour in the Bragg peak region. The efficiency of Gd2O2S:Tb decreases with increasing stopping power (quenching of luminescence) while that of (Zn, Cd)S:Ag increases. Because of these contrasting behaviours it is possible to prepare a mixture of the two scintillating powders in a certain ratio in order to modulate the height of the measured Bragg peak relative to the entrance value so that it is in agreement with the ionization chamber measurements. We propose to use a mixture for the sensitive volume consisting of the following weight fractions: 48% Gd2O2S:Tb, 12% (Zn, Cd)S:Ag and 40% optical cement. PMID- 15552423 TI - On the use of pharmacokinetic models. AB - Extensive use of models in pharmacology, in physiology and in radiotherapy raises some questions on the nature and utility of models in general and of compartmental models in particular. In this paper I will define in a simple and logical way a set of useful pharmacokinetic parameters and show how their estimation depends on the assumed model. A special problem arises when some parameters are not identifiable; in that case I will show how it is possible to determine a range for them. Two examples are used to illustrate how to compute the value of the identifiable parameters and the range of the non-identifiable ones, when the available experimental data are not sufficient to identify a model. PMID- 15552424 TI - Measurement of the three-dimensional distribution of radiation dose in grid therapy. AB - A single large dose of megavoltage x-rays delivered through a grid is currently being utilized by some centres for palliative radiotherapy treatments of large tumours. In this note, we investigate the dosimetry of grid therapy using two dimensional film dosimetry and three-dimensional gel dosimetry. It is shown that the radiation dose is attenuated more rapidly with depth in a grid field than an open field, and that even shielded regions receive approximately 25% of the dose to the unshielded areas. PMID- 15552425 TI - An investigation of the 109Cd gamma-ray induced K-x-ray fluorescence (XRF) bone lead measurement calibration procedure. AB - Two sets of phantoms have been used to calibrate a 109Cd y-ray induced K-XRF bone lead measurement system. Both sets of phantoms are made of plaster of Paris, but the calibration lines are significantly different. This results in a significant difference for the derived concentrations of bone lead for the same person using these two sets of phantoms. This study shows that the different calibration lines are due to the different compositions of the phantoms, which can then be accounted for by adjusting the parameters related to the phantom composition in spectral analysis. Bone-lead concentrations for ten lead-exposed smelter workers were computed before and after analysis modification, and the results show that the bone-lead concentrations for the same person calculated from two sets of phantoms are not significantly different, only after the modifications are incorporated. Through these investigations, it was discovered that a common practice of setting the ratio of the calcium edge amplitude to the coherent scatter amplitude as a constant is only valid when all spectra are acquired at the same system resolution. When there is a change in the resolution between spectra, it has been determined that the ratio of the calcium edge amplitude to the coherent area should instead be used as the constant factor in the analysis program. PMID- 15552426 TI - On the question of 3D seed reconstruction in prostate brachytherapy: the determination of x-ray source and film locations. AB - Inaccuracy in seed placement during permanent prostate implants may lead to significant dosimetric deviations from the intended plan. In two recent publications (Todor et al 2002 Phys. Med. Biol. 47 2031-48, Todor et al 2003 Phys. Med. Biol. 48 1153-71), methodology was described for identifying intraoperatively the positions of seeds already implanted, thus allowing reoptimization of the treatment plan and correcting for such seed misplacement. Seed reconstruction is performed using fluoroscopic images and an important (and non-trivial) component of this approach is the ability to accurately determine the position of the gantry relative to the treatment volume. We describe the methodology for acquiring this information, based on the known geometry of six markers attached to the ultrasound probe. This method does not require the C-arm unit to be isocentric and films can be taken with the gantry set at any arbitrary position. This is significant because the patient positioning on the operating table (in the lithotomy position) restricts the range of angles at which films can be taken to a quite narrow (typically +/- 10 degrees) interval and, as a general rule, the closer the angles the larger the uncertainty in the seed location reconstruction along the direction from the x-ray source to the film. PMID- 15552427 TI - Visible absorption properties of radiation exposed XR type-T radiochromic film. AB - The visible absorption spectra of Gafchromic XR type-T radiochromic film have been investigated to analyse the dosimetry characteristics of the film with visible light densitometers. Common densitometers can use photospectrometry, fluorescent light (broad-band visible), helium neon (632 nm), light emitting diode (LED) or other specific bandwidth spectra. The visible absorption spectra of this film when exposed to photon radiation show peaks at 676 nm and 618 nm at 2 Gy absorbed doses which shift to slightly lower wavelengths (662 nm and 612 nm at 8 Gy absorbed dose) at higher doses. This is similar to previous models of Gafchromic film such as MD-55-2 and HS but XR type-T also includes a large absorption at lower visible wavelengths due to 'yellow' dyes placed within the film to aid with visible recognition of the film exposure level. The yellow dye band pass is produced at approximately 520 nm to 550 nm and absorbs wavelengths lower than this value within the visible spectrum. This accounts for the colour change from yellow to brown through the added absorption in the red wavelengths with radiation exposure. The film produces a relatively high dose sensitivity with up to 0.25 OD units per Gy change at 672 nm at 100 kVp x-ray energy. Variations in dose sensitivity can be achieved by varying wavelength analysis. PMID- 15552428 TI - We need to keep health visiting in the frame. AB - The new NSF on maternity services does not recognise the importance of the health visitor role. PMID- 15552429 TI - Why skin disease needs a higher profile. PMID- 15552430 TI - Doing the knowledge. PMID- 15552431 TI - What plans do you have to improve your IT skills? PMID- 15552432 TI - Seven steps to patient safety. AB - Nurses have a fundamental role in ensuring the safety of patients in their care. Judith Chamberlain-Webber outlines current policy measures aimed at removing the blame culture and encouraging the reporting of incidents and near misses without fear of reprimand, so that lessons can be learned. PMID- 15552433 TI - Rallying Scotland to the cause of health. PMID- 15552434 TI - A point of contact for patients and staff. AB - The concept of nurses working in an extended role was formally introduced by the UKCC 12 years ago, but the role of the rheumatology practitioner had been evolving sporadically over the previous decade. This had resulted in a variety of job titles, including specialist and rheumatology practitioner, with accompanying different levels of resposibility. The changes to junior doctors' hours have meant that more nurses are taking on extended roles. Practitioners have also come from physiotherapy or occupational therapy backgrounds. PMID- 15552435 TI - The challenge of providing spiritual care. AB - Spirituality is an essential element of holistic nursing and nurses need the knowledge and confidence to address spiritual issues with patients. This paper describes the development of a tool to help with broaching this sensitive aspect of care. It argues that nurses need to examine their own spiritual needs in order to be able to help others. PMID- 15552436 TI - Development of a wound-management formulary for use in clinical practice. AB - A wound-management formulary is an important aid to health-care professionals looking for the most appropriate product for a particular patient. Some of the basic principles of compiling such a formulary are described, providing a step-by step guide, and concentrating principally on one for wound dressings. PMID- 15552437 TI - Improving early detection of COPD: the role of spirometry screening assessment. AB - Chronic obstructive pulmonary disease is on the increase, yet public awareness of the condition remains low. Early diagnosis and smoking cessation can arrest the decline in lung status. A primary care-based spirometry screening assessment initiative has improved diagnosis, resulting in early intervention for patients with COPD. PMID- 15552438 TI - Implementing an NHS continence service for nursing home residents. AB - As part of the funded nursing care agenda, the NHS now has responsibility to provide a continence service free of charge for residents of nursing homes. This article explains how such a service was implemented in one PCT. PMID- 15552439 TI - Impact of lower urinary tract symptoms on quality of life and sexual function. AB - Patients with lower urinary tract symptoms may also experience sexual dysfunction, but are reluctant to discuss this with a health-care professional. Nurses who assess these patients have a responsibility to offer non-judgemental support. The investigations and treatment options available are discussed in this article. PMID- 15552440 TI - Choosing the right glove for the right purpose. AB - With the increase in the number of health-care workers developing allergies to natural rubber latex, knowing what alternative gloves can be used is becoming increasingly important. This article provides a guide to the various types of gloves available and their specific uses in health care. PMID- 15552441 TI - The importance of blood pressure control. PMID- 15552442 TI - Changing practice in the subcutaneous infusion of fluids to improve safety. AB - Many nurses administer subcutaneous infusion therapy; it is seen as an alternative to intravenous therapy. But how sharp-safe is the equipment they use? An evaluation of a clinical trial on a catheter to replace metal butterfly needles is outlined, together with details of how this was introduced into clinical areas. PMID- 15552443 TI - Policy development is paramount. Interview by Carolyn Scott. PMID- 15552444 TI - Finding the tools for the job. AB - Steve Kelsey uncovers some of the key challenges facing ward sisters and calls for a more structured development to support the role. PMID- 15552445 TI - Do what I say, not what I do ... PMID- 15552446 TI - Smile for your sphincter. PMID- 15552447 TI - Time to abolish unnecessary male circumcisions. PMID- 15552448 TI - Need to know: vaginal birth after caesarean. Part 1. PMID- 15552449 TI - Water: what are we afraid of? PMID- 15552450 TI - Routine cord blood gas analysis: an overreaction? PMID- 15552451 TI - Litigation and the midwife: shoulder dystocia. PMID- 15552452 TI - Celebrating 10 baby friendly years. PMID- 15552453 TI - Becoming baby friendly. PMID- 15552454 TI - So you think you know how to teach breastfeeding? PMID- 15552455 TI - Midwifery basics: postnatal care. Care of the baby at birth. PMID- 15552456 TI - Topical umbilical cord care at birth. PMID- 15552457 TI - What's on the label? PMID- 15552458 TI - Caught on the hop. PMID- 15552459 TI - Behind the figures. PMID- 15552460 TI - Decision time. PMID- 15552461 TI - Safe admission. Interview by Lynne Pearce. PMID- 15552462 TI - Blood spot screening. Interview by Dina Leifer. PMID- 15552463 TI - Heart of the matter. PMID- 15552464 TI - C is for communication. PMID- 15552465 TI - A world of difference. PMID- 15552466 TI - Test and treat Helicobacter pylori before endoscopy. AB - BACKGROUND: Helicobacter pylori may have major implications for patients' wellbeing and future health. If a patient is found to be H. pylori positive it is important that the infection is eradicated because of the risk of associated peptic ulcers and gastric cancers. There are, however, great demands on NHS gastroenterology and endoscopy services and following the introduction of recent guidelines for dyspepsia some of these issues may be addressed. The literature suggests that a strategy of test and treat before endoscopy referral will benefit patients and be cost-effective. CONCLUSION: There is evidence that, over a period of time, it is more prudent to test and treat H. pylori first and then review the patient's condition before endoscopy is performed (if no other symptoms are identified). PMID- 15552467 TI - Developing and accrediting a CPD framework. AB - This article describes the process to formally accredit existing in-house education courses in a continuing professional development (CPD) framework. The partnership was between two Lothian NHS organisations and a higher education institution (HEI). The nature of the partnership and the differing approaches taken by the organisations are discussed. The authors explore the context of learning in the NHS in relation to the different organisations, and describe the issues identified and the strategies used to address these. The benefits to all stakeholders are listed. Although this case study refers to an initiative in Scotland, the framework model for flexible and work-based education could be adopted across the UK. PMID- 15552468 TI - The management of nausea and vomiting in palliative care. AB - For patients with terminal illness, nausea and vomiting can be very distressing symptoms and can have a negative effect on their quality of life. This article discusses the nursing management of these symptoms, with a view to improving quality of life for these patients. PMID- 15552469 TI - Nursing Standard Nurse 2004 Awards. Time for a change. Interview by Rosalind Bal. PMID- 15552470 TI - Nursing Standard Nurse 2004 Awards. Path finder. Interview by Rosalind Bal. PMID- 15552471 TI - Nursing Standard Nurse 2004 Awards. Running the show. Interview by Rosalind Bal. PMID- 15552472 TI - Nursing Standard Nurse 2004 Awards. Older person's champion. Interview by Dina Leifer. PMID- 15552473 TI - Nursing Standard Nurse 2004 Awards. Up for the challenge. Interview by Ruth Williams. PMID- 15552474 TI - Nursing Standard Nurse 2004 Awards. Home life. Interview by Lynne Wallis. PMID- 15552475 TI - Nursing Standard Nurse 2004 Awards. In perspective. Interview by Steven Black. PMID- 15552476 TI - Nursing Standard Nurse 2004 Awards. Peace of mind. Interview by Steven Black. PMID- 15552477 TI - Nursing Standard Nurse 2004 Awards. Without prejudice. Interview by Ruth Williams. PMID- 15552478 TI - Nursing Standard Nurse 2004 Awards. Breathe easy. Interview by Steven Black. PMID- 15552479 TI - Nursing Standard Nurse 2004 Awards. Closer to home. Interview by Lynne Wallis. PMID- 15552480 TI - Future sights. PMID- 15552481 TI - Will the bank make us or break us? PMID- 15552482 TI - Is there too much qualitative nursing research? PMID- 15552484 TI - How to work abroad. PMID- 15552485 TI - Sorted: protecting the gay community. Interview by Janis Smy. PMID- 15552486 TI - 'Daniel taught me lessons I'll remember forever'. PMID- 15552487 TI - Measuring the impact of modern matrons in the ward setting. AB - With the The NHS Plan (Department of Health, 2000) the government made it clear there was a need to reintroduce the role of matron to establish a clear responsibility for standards in the ward setting. The role of modern matron is now considered to be a fundamental part of clinical governance, ensuring the quality of the care provided is of the highest standard. Staff at South Essex Partnership NHS Trust instituted an audit to provide an accurate picture of the impact of modern matrons. PMID- 15552488 TI - What you need to know about rabies. PMID- 15552489 TI - H2-receptor antagonists. PMID- 15552490 TI - Multidisciplinary meetings in medical admissions units. AB - Multidisciplinary working was introduced in a medical admissions unit to improve the discharge process. The number of patients discharged increased as did the opportunities for discharge because they took place throughout the week including weekends. The number of medical patients occupying surgical beds decreased by 100 per cent, which reduced disruption to elective surgery schedules. PMID- 15552491 TI - Incidence, pathophysiology and treatment of cervical cancer. AB - Cervical cancer is the third most common cancer affecting women. While not in itself a sexually transmitted disease, cervical cancer is linked to the presence of the human papilloma virus, which is sexually transmitted. Survival rates are significantly higher where diagnosis is early, and a national screening programme has reduced mortality in the UK. However, uptake is low in some groups and nurses have a role in increasing uptake. PMID- 15552492 TI - A study into safe and efficient use of defibrillators by nurses. AB - AIM: To find out if ward nurses can perform rapid defibrillation at six months and one year after they have completed training in this procedure. METHOD: A longitudinal quasi-experimental study. RESULTS: Fifteen registered nurses and one enrolled nurse demonstrated rapid and safe defibrillation at six months and one year after training. CONCLUSION: Ward nurses who have a range of qualifications and experience can deliver safe and rapid defibrillation. PMID- 15552493 TI - Understanding the importance of effective glycaemic control. PMID- 15552494 TI - How to improve care through effective decision-making. PMID- 15552495 TI - Trusts take control of staff training. PMID- 15552496 TI - Stroke: fact and fiction. PMID- 15552497 TI - Seeing beyond adversity. PMID- 15552498 TI - Male carers: myth or reality? PMID- 15552499 TI - Into the unknown: establishing a new service in an acute hospital setting. PMID- 15552500 TI - Rehabilitation: more than activities of daily living. PMID- 15552501 TI - Developing and maintaining a therapeutic relationship: Part 1. PMID- 15552502 TI - Assessing the cultural and religious needs of older people. AB - Patients' cultural and religious needs can become particularly important during ill health. This article describes two different frameworks and how they can be used by nurses to assess those needs among older people in their care. PMID- 15552503 TI - Antiplatelet therapy. PMID- 15552504 TI - Mental health in acute care: Part two. PMID- 15552505 TI - Becoming invisible. PMID- 15552506 TI - Heart of the matter. PMID- 15552507 TI - My husband and I. PMID- 15552508 TI - Benefit/risk of therapies for rheumatoid arthritis: underestimation of the "side effects" or risks of RA leads to underestimation of the benefit/risk of therapies. AB - Most physicians are familiar with the side effects or risks of drugs used to treat rheumatoid arthritis (RA), but relatively less familiar with the "side effects" or risks associated with RA itself RA is not thought to have the same potential severity as a cardiovascular or neoplastic disease by most physicians, the public, or even some rheumatologists, although relative rates of predicted mortality in some patients with RA are in the range of some people with coronary artery disease or Hodgkin's disease. Many reasons may be identified to explain why the risks of RA have been underestimated: RA does not lead to acute life threatening situations; population-based data have suggested that most people who meet criteria for RA have a mild or self-limited process; acute attributed causes of death in people with RA are superficially similar to those in the general population; clinical trials have suggested many therapies that are efficacious over a period of 3-12 months; few long-term longitudinal studies were performed prior to the 1980s; medical recommendations made during the 1950s-1980s suggested that simple therapies were adequate for most patients; and quantitative information concerning patient status is generally not included in standard rheumatology care. As more information has emerged concerning severe long-term outcomes in the "natural history" of RA (as treated prior to the 1990s), new strategies of aggressive intervention have been developed. Furthermore, basic research has led to new therapies. It appears that the benefit/risk ratio of therapies for RA has increased substantially over the last two decades, and the outlook for patients with RA is much better at this time than in previous years. PMID- 15552509 TI - Undifferentiated arthritis--disease course assessed in several inception cohorts. AB - The prognosis of patients with undifferentiated arthritis (UA) may vary from self limited to severe destructive rheumatoid arthritis (RA). Because early aggressive treatment might offer an effective means to slow disease progression in RA, it is important to identify UA patients who will develop RA and treat them as early as possible. At the same time, inappropriate treatment of patients with a more benign disease course should be avoided. Here, an overview is given of the characteristics and numbers of patients with UA who evolve into RA. UA is defined as any arthritis that has the potential for a persistent course, without fulfilling the classification criteria for specific rheumatic disorders. To compare endpoints in the different databases, the 1987 ACR criteria for RA were used. In the nine databases employing a similar definition for undifferentiated arthritis, the proportion of patients with UA that evolved into RA within 1 year varied from 6% to 55%. These differences arise in large part from differences in the inclusion criteria and in the definitions used for UA and RA. The data from the various cohorts support a hypothesis that a considerable proportion of UA patients are actually patients with RA in a very early stage. Controlled intervention studies with early antirheumatic treatment in these patients are mandatory in order to provide further insight into the natural course of UA and to define a treatment strategy that will successfully slow or prevent disease progression. PMID- 15552510 TI - New approaches to imaging early inflammatory arthritis. AB - Imaging techniques such as musculoskeletal ultrasonography (MUS) and magnetic resonance imaging (MRI) are playing an increasingly important role in the assessment of patients with inflammatory arthritis. Such modalities are now used routinely in the evaluation of joint, tendon and soft tissue inflammation and bone damage in many early arthritis clinics. They have the ability to directly visualise, characterise and quantify the earliest inflammatory changes and have proved not only to be useful additional complimentary clinical tools to improve the speed and accuracy of diagnosis, direct appropriate treatment, monitor response to therapy, measure disease progression and outcome but also continue to contribute to our understanding of disease pathogenesis. These imaging methods may therefore offer a significant advantage as they endorse the principles of early diagnosis and optimal targeted therapy essential to providing the most favourable long term outcome for patients with inflammatory arthritis. This article reviews the current evidence supporting the role of MUS and MRI in the assessment of patients with inflammatory arthritis. PMID- 15552511 TI - Quantitative documentation of benefit/risk of new therapies for rheumatoid arthritis: patient questionnaires as an optimal measure in standard care. AB - Assessment of benefit/risk of therapies for any disease is best conducted according to quantitative data. In many diseases, such as hypertension or hyperlipidemia, a single quantitative measure serves as a "gold standard" for patient status, but no single measure can serve as a "gold standard' for all individual patients with rheumatoid arthritis (RA). Therefore, indices such as the American College of Rheumatology (ACR) Core Data Set and Disease Activity Score (DAS), are used in clinical trials and other clinical research. These indices include 3 types of measures, which are derived from a health professional [joint counts, global]; a laboratory [erythrocyte sedimentation rate (ESR) and C reactive protein (CRP)]; or a patient questionnaire [physical function, pain, global]. In most standard clinical care, the majority of clinicians do not collect joint count or patient questionnaire data at most visits. Therefore, assessment and management of most patients with RA is conducted empirically, with the only quantitative data from laboratory tests. Measures on a patient self report questionnaire of physical function, pain, and global status, are as informative as joint counts, radiographic scores, laboratory tests, or any measure by a health professional to document status, estimate prognosis, and monitor responses to therapies. We suggest that quantitative measurement may be incorporated into standard clinical care most easily and effectively by asking each patient to complete a simple 1-page questionnaire at each visit to a rheumatologist. PMID- 15552512 TI - Internet-based monitoring of patients with rheumatoid arthritis. AB - Patient-derived measures have been increasingly recognized as a valuable means for monitoirng patients with rheumatoid arthritis. One advantage of this data is that it can be collected remotely. This would allow more frequent and more rapid assessments, which could optimize therapeutic intervention and patient outcome. PMID- 15552513 TI - Mandatory pharmacosurveillance--a Canadian model for access to therapy and research. AB - Regulatory authorities in Canada have expressed a vital need for pharmacoepidemiological data on long-term effectiveness, safety, and cost-benefit of new therapies, particularly in comparison to currently available therapies, in routine clinical practice to allow informed decision making in listing new therapies on formulary. We describe the evolution of a new model of pharmacosurveillance involving a partnership between academic and community rheumatologists, government, and industry whereby access to therapy is conditional on participation in an industry-funded pharmacosurveillance study that assesses long-term effectiveness, safety, and cost-benefit. Though funded by industry, the program is administered by government and designed and operated at arms length from industry. The clinic data sheets are available at www. altarheum.com. The program also provides a sustainable model for promoting observational research on therapeutics in general. PMID- 15552514 TI - Healing of erosive changes in rheumatoid arthritis. AB - This is an overview over the history and present state of knowledge of radiographic signs of erosion healing. The existence of healing or repair has been confirmed; different observers agree in the identification of healing; it may be identified without knowing the sequence of the films. As healing indicates that inflammation has discontinued for several months in an individual joint, it might represent a good additional outcome measure in RA clinical trials. PMID- 15552515 TI - Relative versus absolute goals of therapies for RA: ACR 20 or ACR 50 responses versus target values for "near remission" of DAS or single measures. AB - Therapies for rheumatoid arthritis (RA) may be assessed according to relative levels of measures to compare efficacy to another therapy or to a placebo, as in the American College of Rheumatology (ACR) 20%, 50%, or 70% (ACR 20 ACR 50 and ACR 70) responses, or by absolute levels of measures, as in disease activity scores (DAS), ACR criteria for remission, or "target values" of specific measures. Regulatory considerations have emphasized primarily relative comparisons to a placebo or standard therapy, derived in part from the weak efficacy of traditional disease modifying anti-rheumatic drugs (DMARDs). While improvement compared to placebo certainly indicates efficacy, it is of concern that measures of inflammatory activity, such as swollen joints and the erythrocyte sedimentation rate (ESR), may be stable or improved over periods of 5 10 years, while measures of damage, such as joint deformity and radiographic changes, may progress over the same period in the same patients. These findings suggest that improvement at a level of 20% or 50% may deter but not prevent severe long-term outcomes of radiographic progression, functional declines, work disability, and premature mortality, seen in most patients until the middle 1990s. Outcomes appear to be improved at this time, associated with aggressive treatment strategies and more powerful therapies, including biologic agents. In the Finnish Rheumatoid Arthritis Combination Therapy Trial (FinRACo), no patient who was in remission after 6 months was receiving work disability payments 4 1/2 years later, compared to 22% of patients who had ACR 20 or 50 responses and 54% of patients who did not have ACR 20 responses after 6 months who were all receiving work disability payments after 5 years. These findings suggest that absolute targets, including remission, may be realistic contemporary goals, with aggressive treatment strategies and more effective DMARDs and biologic agents. PMID- 15552516 TI - Longer term benefits of treating rheumatoid arthritis: assessment of radiographic damage and physical function in clinical trials. AB - Evidence from randomized controlled trials with newly approved DMARD therapies have proven efficacy by American College of Rheumatology [ACR] response criteria, Disease Activity Scores [DAS] and radiographic measures of disease progression. Treatment over 2 years duration results in clinically meaningful improvements in physical function, by the Health Assessment Questionnaire [HAQ] and health related quality of life [HRQOL], using the medical outcomes short form 36 [SF 36]. Changes in HAQ are evident within one month, maximal at 3-6 months, and sustained over 24 months, reflected by improvements in social functioning, role emotional and the general health profile as well as physical domains of SF-36. Trials with the new DMARDs, as well as MTX, indicate that long term benefits in radiographic damage and physical function can be inferred from treatment data over 12 months. "Successful" patients who continue to do well will derive benefit from treatment for as long as 3 to 5 years. Recent RCTs indicate that combination therapy, initiated together, offers more improvement in radiographic progression and physical function than monotherapy, although the trial data cannot yet tell us which DMARD may be most beneficial in a given patient. Once treatment with any one agent--biologic or synthetic--is initiated, the addition of a second agent should occur rapidly, if for example active disease persists 8 or 12 weeks later, without waiting for documented treatment failure. If this treatment paradigm is followed, along with regular assessments of radiographic damage and physical function, then patients may more likely derive long term clinical benefit than with traditional approaches. PMID- 15552517 TI - Cost-effectiveness of anti-tumor necrosis factor agents. AB - Rheumatoid arthritis can lead to substantial morbidity, disability and mortality. The development of anti-tumor necrosis factor antibodies from the bench to the bedside over the past 15 years has ushered in the new era of biologic therapies for rheumatic diseases. Etanercept, infliximab and adalimumab have all been approved for the treatment of rheumatoid arthritis on the basis of improved clinical outcomes. Because these treatments, however, are expensive and not uniformly effective, concerns have arisen regarding their cost-effectiveness. This paper reviews the disease burden of rheumatoid arthritis, costs of drug therapy, costs of rheumatoid arthritis and the economics and cost-effectiveness of anti-tumor necrosis factor antibody agents. PMID- 15552518 TI - Benefit/risk of combination therapies. AB - It is now accepted that rheumatoid arthritis is not a benign disease, and has considerable morbidity and increased mortality rates. Monotherapy with disease modifying anti-rheumatic drugs (DMARDs) is often ineffective, and rarely leads to sustained clinical remission. Many clinical trials suggest the effectiveness of using combination therapies, and also the benefit of aggressive management early in the course of the disease. However, recent publications studying a variety of combination therapies in rheumatoid arthritis have shown diverse results. Drug combinations at an early stage of rheumatoid arthritis which slow radiographic progression appear to be the most convincing. These data suggest that practitioners should begin with intensive therapy in early disease, and not reserve combination therapy for those who fail monotherapy. The therapeutic strategy should be positioned according to the severity of the disease: in RA with markers indicating severity, combination should be initiated at the start of therapy while it should be instituted in a rapid step-up fashion in mild RA if insufficient efficacy is seen for monotherapy. In general, combinations appear safe and well-tolerated, but continued caution with appropriate monitoring of long-term results and possible toxicities is required. PMID- 15552519 TI - Glucocorticoid use in rheumatoid arthritis: benefits, mechanisms, and risks. AB - Glucocorticoids have long been recognized to have beneficial effects in rheumatoid arthritis (RA) (1,2). Several clinical trials over the last decade have further documented the efficacy of glucocorticoids in relieving inflammation and in preventing radiographic erosions in early RA (3-5). Additionally, research has yielded new insights about the cellular mechanisms responsible for these perceived beneficial effects (6,7). Despite potential short term benefits, there is a lack of demonstrated long-term efficacy as well as concerns about short and long-term toxicity. Although these concerns have limited enthusiasm for glucocorticoids by many patients and practitioners, in the U.S. it is estimated that 44% to 75% of RA patients use glucocorticoids (8,9). Confusion and controversy may relate to the fact that toxicity reports are also limited by only modest data quality and quantity. Given growing clinical and basic science evidence supporting the efficacy of glucocorticoids for the treatment of rheumatoid arthritis, their use may further increase. In this review we will examine the latest data supporting the benefits and risks of glucocorticoid use in RA. PMID- 15552520 TI - Benefit and risk of methotrexate treatment in rheumatoid arthritis. AB - This is a literature review on the efficacy and toxicity of low dose weekly methotrexate treatment in rheumatoid arthritis. Personal recommendations on dosing and monitoring (of) the drug are given. PMID- 15552521 TI - Benefit/risk of leflunomide in rheumatoid arthritis. AB - Leflunomide was first shown to have disease-modifying properties in a rat model of adjuvant-induced arthritis. Leflunomide has been subsequently used with success in several animal models of tissue and organ allograft and of autoimmune disease including collagen- and adjuvant-induced arthritis, interstitial nephritis, myasthenia gravis, and systemic lupus erythematosus. Based on its success as an immunosuppressive agent in these models, leflunomide was tested for the treatment of rheumatoid arthritis (RA). PMID- 15552522 TI - Benefit/risk of cyclosporine in rheumatoid arthritis. AB - Combination therapy has emerged as a crucial therapeutic tool to control aggressive rheumatoid arthritis (RA). Cyclosporine (CsA) when combined with methotrexate (MTX) has shown substantial benefit in clinical practice. The primary benefit is its positive effect in the control of joint-bone erosions. The most feared adverse effect is the development of nephrotoxicity, which may be in part hemodynamic and in part structural, i.e. fibrotic. Careful monitoring of concomitant drugs, hypertension and through blood levels should allow the patient to maintain normal renal function. The successful employment of CsA in lupus nephritis clearly supports this statement. PMID- 15552523 TI - Overview of benefit/risk of biological agents. AB - Targeted tumor necrosis factor-alpha antagonists, first approved by the FDA in 1998, have had a significant impact on the treatment of patients with rheumatoid arthritis. In general, the benefit/ risk ratio for these agents and the IL-1 receptor antagonist, anakinra, has been quite favorable. However, infrequent adverse events can be serious and require continued pharmacovigilance. Infections, particularly tuberculosis and less commonly fungal infections, are among the most serious adverse events, especially given delays in diagnosis due to subtle or atypical presentations. Questions have also arisen regarding whether anti-TNF-alpha agents increase the risk of lymphoma, a complicated issue confounded by the multiple risk factors for lymphoma in patients with rheumatoid arthritis and low observed incidence rates of lymphoma, requiring prolonged monitoring. Additional rare reported complications include systemic lupus erythematosus-like syndromes, congestive heart failure and demyelinating syndromes (including cases resembling progressive multifocal leukoencephalopathy). Ongoing post-marketing surveillance of these and other serious adverse events is necessary to determine the true incidence rates, and whether a reassessment of the overall risk-benefit of tumor necrosis factor-alpha antagonists will be required. PMID- 15552524 TI - Switching between biological agents. AB - The TNF-alpha antagonists infliximab, etanercept and adalimumab have similar efficacies in clinical trials in the rheumatic diseases, and this efficacy may be related primarily to their neutralizing free TNF-alpha. Thus, a reasonable question for clinicians is whether patients who have failed one TNF-alpha antagonist could reasonably be given a trial with another such agent, or whether this is simply a waste of time and money. Several published studies have addressed this important practical issue and are reviewed in this paper. Data from the Stockholm TNF-alpha follow-up registry "STURE" that address this issue are described in detail. The overall conclusion appears to be that such switches of biologicals can be effective. Nonetheless, further attention should be paid to the details of various clinical scenarios in which this question can arise and the methods by which comparisons are made of treatment effects occurring during sequential therapies. PMID- 15552525 TI - Benefits and risks of biological agents: lymphomas. AB - Lymphomas are uncommon malignancies of unknown aetiology. Rheumatoid arthritis is a known risk factor for lymphoma, and some studies show that this risk is higher in patients with more severe disease. The causes of the association between RA and lymphoma are not understood. Conventional anti-rheumatic agents may increase the risk for lymphoma, but these associations are relatively weak at most. For the currently available TNF-alpha antagonists, available data include the possibility of a somewhat higher risk for lymphoma than for patients not treated with such agents, but also point to several sources of bias that could explain a possible association. Current practice recommendations should probably not go further than an awareness of the possibility of lymphoma in any patient with RA exhibiting unexplained systemic symptoms. PMID- 15552526 TI - Tuberculosis and opportunistic infections: relevance to biologic agents. AB - With the expanding use of biologic agents, in particular TNF inhibitors, tuberculosis and other opportunistic infections have become an important and growing concern in rheumatology. Clinicians using these therapies should have an understanding of the scope of the problem, the underlying scientific rationale, as well as the optimal approaches to screening, monitoring and treatment. PMID- 15552527 TI - Demyelination and inhibition of tumor necrosis factor (TNF). AB - The development of tumor necrosis factor alpha (TNFalpha) inhibitor therapy is arguably the most significant achievement in the treatment of rheumatic diseases to date. One serious potential side effect associated with these agents is the onset of neurologic signs and symptoms. In this paper we will examine the relationship of TNFalpha antagonism and demyelinating disease. Reviewing early laboratory and animal models, we discuss the mechanism of TNFalpha in central nervous system (CNS) injury and repair. Two negative studies of TNFa inhibitor therapy in the treatment of refractory multiple sclerosis (MS) are considered. From the manufacturers' clinical development programs and post-marketing adverse event reporting data, we report the current incidence of demyelinating symptoms associated with each of the commercially available anti-TNFa agents. Comparing these reports to the incidence of MS in society as a whole, we find the rate of new cases of neurologic disease in exposed patients is not different from the rate of expected cases. Finally we explore arguments that support and refute a potential biologic relationship between TNFa neutralization and demyelinating disease. PMID- 15552528 TI - Unusual toxicities with TNF inhibition: heart failure and drug-induced lupus. AB - Serious and unexpected adverse events, such as heart failure and drug-induced lupus, have been reported in patients receiving TNF inhibitor therapy. These events generally are easily recognizable, although they cannot be predicted nor avoided, other than by drug avoidance altogether Many patients have great benefit from anti-TNF therapies. Their intelligent use requires a firm understanding of these rare toxicities, so as to minimize the morbidity associated with their uncommon occurrence. PMID- 15552529 TI - Safety issues related to emerging therapies for rheumatoid arthritis. AB - Several novel biologic therapies for rheumatoid arthritis (RA) are emerging that target aspects of the immune system other than tumor necrosis factor (TNF). Two such therapies currently in development include CTLA4Ig (Abatacept) and anti-CD20 monoclonal antibody (Rituximab). CTLA4Ig has been demonstrated to be well tolerated with a good short-term safety profile. Rituximab has also been shown to have a good safety profile in a limited number of RA patients. Further safety data with larger numbers of RA patients treated with Rituximab is required. PMID- 15552531 TI - [Morphometric variability of precuneus in relation to gender and the hemisphere of human brain]. AB - Precuneus, a quadrangular gyms of the medial surface of the human parietal lobe, is bound by three primary brain sulci and by superior hemispheric border. Precise encephalometric data about precuneus are important in the studies of brain lateralization, sex dimorphism, and brain functions in general. In this study, total and visible (exstrasulcal) surface area of the precuneus were measured on 50 brains of the adult persons (31 male, and 29 female), together with the investigation of its relationship to the side (left/right) and gender (sex dimorphism). The average total surface area of the precuneus was 16.07 cm2 on the right (males 16.44 cm2, females 15.27 cm2), and 15.44 cm2 on the left (males 15.67 cm2, females 14.62 cm2). The average visible (extrasulcal) surface area of cortex of precuneus was 9.97 cm2 on the left (males 10.75 cm2, females 8.91 cm2), and 9.38 cm2 on the right (males 10.25 cm2, females 8.19 cm2). Exstrasulcal surface area of the left precuneus was larger, by 0.59 cm on the average, which was not statistically significant. Total surface area of precuneus of males was significantly larger on the right (16.44 cm2) (p<0.01) than on the left (15.67 cm2). In females it was also larger on the right (15.27 cm2) than on the left (14.62 cm2), but with no statistical significance. Visible (exstrasulcal) surface area of both, (left and right precuneus of males), was highly significantly larger in comparison with the females (p<0.001). The obtained results and other facts suggested that sex dimorphism of human brain, including precuneus, was present, but not always easily observable, studied or proven in all the details. PMID- 15552530 TI - Presence of interleukin-8 and the IL-1 receptor antagonist in the cervical mucus of fertile and infertile women. AB - BACKGROUND: Cytokines are involved in almost every aspect of reproduction, and recent studies suggested a relationship between cytokines and male/female infertility. In the present study, the levels of interleukin-8 (IL-8) and interleukin-1 receptor antagonist (IL-1Ra) were determined in the cervical mucus of fertile and infertile women. METHODS: Groups of patients were formed according to the results of the standard procedure for infertility investigation, including postcoital test and the presence of antispermatozoid antibodies in the sera of both partners and in seminal plasma, by mixed antiglobulin reaction (MAR) test and Kibrick agglutination test. IL-8 and IL-1Ra levels were determined in solubilized (ultrasonographic sonication) cervical mucus sampled in the midcycle by commercial ELISA kits and expressed as pg/mg proteins. RESULTS: The groups were designated as fertile (n=20) and infertile (n=48). The latter was divided into two subgroups, one consisting of infertile women with positive postcoital test and without antispermatozoid antibodies (n=30), and the other designated as infertile women with negative postcoital test (n=18). This subgroup was composed of women with negative postcoital test and without antibodies (n=10) and the women with negative postcoital test with antibodies (n=8). Similar levels of IL-8 and IL-1Ra were noted in the cervical mucus of infertile women and women with positive postcoital test and without antispermatozoid antibodies. A tendency of decrease (p=0.052) and significant decrease in IL-8 levels (p<0.05) was noted in negative postcoital test group and negative postcoital test group without antibodies, respectively, compared to the levels in the fertile examinees. A significant rise in IL-1Ra levels (p<0.05) was detected in the mucus of negative postcoital test group with further increase in negative postcoital test group with antibodies (p<0.02). CONCLUSION: Changes in IL-8 and IL-1Ra levels in the cervical mucus of infertile patients with negative postcoital test suggested the existence of the relationship between cervical cytokines and infertility in these women. PMID- 15552532 TI - [Myocardial perfusion scintigraphy in identification and localization of individual coronary lesions]. AB - The aim of this study was to evaluate the accuracy of myocardial perfusion scintigraphy with technetium-99m tetrofosmin by single-photon emission computed tomography (SPECT), using one-day protocol in the identification and localization of individual stenosed coronary vessels. Sixty-eight patients with suspected coronary artery disease (CAD) were studied. In thirty of them coronary angiography showed significant stenosis (> or = 50%). Nine patients were with one vessel disease, 11 were with two-vessel disease, and 10 were with three-vessel disease. All the patients were administered two i.v. injections of 99mTc tetrofosmin, one at peak pharmacologic exercise (1-3 min after i.v. administration of dipiridamol 0.56 mg per kg during 4 min) 370 MBq, and the other 740 MBq at rest 3 hrs after the exercise test (acquisition was obtained 15-30 min after injections for both studies). Overall sensitivity, specificity, and diagnostic accuracy in the identification of individual stenosed coronary vessels were 90%, 86%, and 88%, respectively. Sensitivity, specificity, and accuracy in each of the individual vascular territories were not significantly different: LAD (96%, 64%, and 75%), ACx (73%, 100%, and 94%), RCA (95%, 93%, and 94%). The results of this study demonstrated one-day 99mTc-tetrofosmin SPECT scintigraphy to be suitable and accurate technique for the identification and localization of individual stenosed coronary vessels, as well as a highly sensitive method in the recognition of one- and multiple-vessel diseases of coronary arteries. PMID- 15552533 TI - [Assessment of diet therapy effect on blood pressure of obese persons with and without antihypertensive therapy]. AB - BACKGROUND: Arterial hypertension is a multicausal chronic disease often accompanied by obesity. The aim of this investigation was to examine the effect of diet therapy in the treatment of obese hypertensives with and without antihypertensive therapy. METHODS: The investigation was conducted at the Department of Nutrition on a sample of 110 obese hypertensive patients on diet therapy. Subjects were divided into two groups: the group on diet therapy with antihypertensive drugs E1 (n=78), and the group on diet therapy without pharmacotherapy E2 (n=32). Nourishment state i.e., obesity level was assessed by body mass index (BMI). All the patients belonged to the obese group--BMI > 30 kg/m2. Besides blood pressure values, the following parameters were monitored: serum cholesterol, trygliceride levels and BMI. RESULTS: The obtained results in the group with diet therapy combined with antihypertensive therapy showed highly significant decrease of anthropometrical parameters: body weight (99.14 kg vs. 90.16 kg) (p<0.001) and BMI (37.32 vs. 34.09) (p<0.001); percent body fat (41.97 vs. 38.78) (p<0.001); systolic (154.81 vs. 141.91) (p<0.001), and diastolic pressures (95.42 vs. 87.36) (p<0.001); cholesterol (6.39 vs. 5.99) (p<0.002), and triglycerides concentration (2.69 vs. 2.21) (p<0.019). In the group on single diet therapy, highly significant decrease of body mass (99.33 vs. 90.18) (p<0.001), BMI (34.79 vs. 31.58) (p<0.001), percent body fat (39.27 vs. 36.70) (p<0.001), systolic (148.44 vs. 132.74) (p<0.001), and diastolic pressures (93.97 vs. 82.90) (p<0.001), was achieved, while the differences between initial and final cholesterol and triglyceride concentrations, although observed, were not statistically significant. CONCLUSION: The obtained results implicated that diet therapy significantly helped the normoregulation of both systolic and diastolic blood pressure. Considering this, during physicians' routine practice in the treatment of hypertension, attention should be paid on the reduction of the corresponding level of obesity. PMID- 15552535 TI - [The frequency of resistance to antibiotics of most frequently isolated bacteria from blood cultures during the period 1997-2002]. AB - The aim of this study was to determine the frequency of resistance to antibiotics of the most frequently isolated bacteria from blood cultures of hospitalized patients during the period 1997-2002. The resistance to antibiotics was determined by disk diffusion method according to National Committee for Clinical Laboratory Standards procedures. The majority of staphylococci isolates were resistant to methicillin, and the proportion of methicillin-resistant Staphylococcus aureus was stable (76.8-81.6%), during the follow-up period. None of the staphylococci isolates were resistant to vancomycin, but there was a very high incidence of high-level resistance of enterococci to aminoglycosides (47.2 72.2%). In 1998, only one strain among enterococci was resistant to vancomycin (Enterococcus faecium, VanA phenotype). Enterococcus spp isolates expressed variable frequency of resistance to ampicillin (15-40.1%) during the follow-up period. Among Enterobacteriaceae there were no isolates resistant to imipenem, but dramatic increase of the resistance to ceftriaxone was found from 35.9% in 1997 to 95.9% in 2002 (p<0.001). Extended spectrum beta-lactamases production was found in all the species of enterobacteria isolates. Resistance to imipenem was observed in Acinetobacter spp isolates in 2002 for the first time. Pseudomonas spp isolates expressed high and very variable resistance to all antibiotics tested during the follow-up period. PMID- 15552534 TI - [Occupational exposure to ionizing radiation and the occurrence of cataract]. AB - Radiation cataract is one of ensuing effects of ionizing radiation, since its threshold dose under which it does not occur, and above which it shows dose dependency, has been observed. Clinical course of radiation cataract is identical for all the types of ionizing radiation and is very typical. Minimal dose for progressive cataract formation is determined by the type of radiation, i.e., its relative biological efficacy, dose, and the duration of the exposure period. Theoretically, threshold dose existence does not exclude the incidence of cataract formation under significantly smaller doses, as well. The aim of this study was to analyze the incidence of cataract formation among the medical staff professionally exposed to ionizing radiation. Neither of the diagnosed cataracts had typical morphology, nor was the correlation established between the dose, exposure time, and the cataract formation. All the diagnosed cataracts were described as premature, and therefore ionizing radiation was considered as a co factor in premature cataract formation in the examined groups. PMID- 15552536 TI - Carbamazepine for acute psychosis with EEG abnormalities. AB - AIM: To investigate the efficacy of carbamazepine as adjuvant drug therapy in acute paranoid psychosis with associated EEG abnormalities, compared to sole antipsychotic treatment. METHODS: Eleven medication-naive patients, diagnosed with acute paranoid psychosis with associated EEG abnormalities, were divided into two treatment groups: sole fluphenazine group, with flexible dosing of 5-10 mg/day (n=6), and carbamazepine group (n=5) with the addition of carbamazepine (600 mg/day) to fluphenazine treatment. Clinical Global Impression (CGI), Brief Psychiatric Rating Scale (BPRS), Scale for the Assessment of Negative Symptoms (SANS), and EEG were assessed on the baseline and after 6 weeks of treatment. Paired and two-tailed t-tests were used for statistical significance. RESULTS: All the patients showed significant improvement of mental state after 6 weeks of treatment with no significant differences in CGI, BPRS, and total SANS scores in relation to the therapy with carbamazepine. Nevertheless, after 6 weeks of the treatment, EEG findings were significantly better in carbamazepine group, in relation to the findings from the onset of the treatment, as well as in comparison to sole fluphenazine group. CONCLUSION: Although carbamazepine stabilized abnormal brain electrical activities it seemed that the associated EEG abnormalities were not significant for acute psychosis observed. These preliminary results suggested that there was no convincing evidence that carbamazepine was efficient as the augmentation of antipsychotic treatment for patients with both acute paranoid psychosis and EEG abnormalities. PMID- 15552537 TI - [The role of Helicobacter pylori infection in the development of gastric neoplasms]. PMID- 15552538 TI - [Opioid analgesics]. PMID- 15552539 TI - [Measures for the improvement of medical practice]. PMID- 15552540 TI - [Aspiration syndrome]. PMID- 15552541 TI - [Transitional type of Castleman's disease manifested as the POEMS syndrome]. AB - BACKGROUND: Castleman's disease is an atypical lymphoproliferative disorder characterized by angiofollicular hyperplasia of lymph nodes. Histologically, it can be classified into a hyaline-vascular type, plasma-cell type, and transitional (mixed-cell) type, while clinically localized type has been classified as unicentric, or generalized (multicentric) form of the disease. CASE REPORT: This paper presents a 21 year old male patient with multicentric Castleman's disease, a transitional (mixed-cell) type. The disease was manifested by fever, generalized enlargement of peripheral lymph nodes, breast glands enlargement, hyperemia of the face, and weakness of the lower extremities. We found mediastinal lymphadenopathy, pleural and pericardial effusions, sensorimotor peripheral neuropathy and polyclonal hypergammaglobulinemia. The simultaneous presence of these manifestations of the disease (sensomotor peripheral neuropathy, lymphadenopathy, effusions, endocrinopathy, polyclonal gammaglobulinemia and skin changes) is indentified as POEMS syndrome. The diagnosis of Castleman's disease was based on the results of histopathologic analysis of mediastinal lymph node biopsies after thoracotomy. The patient was treated with corticosteroids (prednisone 80 mg daily for 2 weeks followed by 60 mg daily). A partial response was achieved after 4 months of treatment. CONCLUSION: A transitional type of multicentric Castleman's disease may be present itself as POEMS syndrome. The effect of corticosteroid therapy in this form of the disease is unpredictable. PMID- 15552542 TI - [Subcutaneous emphysema as a complication of tooth extraction]. AB - Subcutaneous emphysema is a rare complication in dentistry, which may lead to diagnostic errors and inadequate therapy. A 17 year old female patient, in whom the separation of tooth roots was performed by the use of air-powered drill during the extraction of the first right lower molar, is presented in this paper. During the intervention, swelling of the right half of the face and the lower eyelid suddenly occurred, accompanied with simultaneous feeling of choking, and pressure in the neck and chest. Because of the suspicion of the allergic reaction, the patient was administered antihistaminic agent, together with parenteral corticosteroid, and was sent to an institution where she was treated as an in-patient. During hospitalization, subcutaneous emphysema of the face and neck was diagnosed by physical and x-ray examination. The emphysema completely disappeared after the use of oxygen and antibiotics. PMID- 15552543 TI - Cognitive-behavioral treatment for panic disorder: current status. AB - Is cognitive behavioral treatment (CBT) appropriate for panic disorder with or without agoraphobia (PDA) in children, adolescents, and adults? Are its effects durable? In this review, we survey various psychological approaches to the treatment of PDA and examine the relative efficacy and clinical utility of each. A growing body of research demonstrates that CBT is well-tolerated, cost effective, and produces substantial treatment gains for individuals with PDA over the short- and long-term. Nevertheless, not everyone benefits and there is room for improvement among those who do. We address these shortcomings and consider recent developments. PMID- 15552544 TI - No significant QTc interval changes with high-dose ziprasidone: a case series. AB - This study examined the effects of high-dose ziprasidone on the electrocardiograms (ECGs) of adult inpatients. This was done in an effort to assess the incremental risk of QTc interval prolongation when using ziprasidone in doses above the maximum dose of 160 mg/day approved by the Food and Drug Administration. We retrospectively examined pre- and post-treatment QTc intervals in 15 subjects at the James A. Haley Veterans Affairs Medical Center who had received high doses of ziprasidone, ranging from 240 to 320 mg/day, due to intractable psychotic symptoms. Pure baseline ECGs were not always attainable, since the majority of the subjects were taking concomitant medications at the time that ziprasidone was initiated. Analysis yielded an average increase of 3.4 msec from pre- to post-treatment, with a maximum post-treatment interval of 452 msec and no cases having a pre- to post- treatment QTc interval increase > 20 msec. These findings are statistically insignificant and do not approach the 500 msec threshold of concern. This study suggests that high-dose ziprasidone does not have a significant incremental effect on QTc interval prolongation in certain subsets of patients. However, it does not rule out the possibility that ziprasidone could have deleterious effects in higher risk populations or that ziprasidone could have rare, idiosyncratic cardiogenic effects. Limitations of the study are identified and recommendations for future research are presented. PMID- 15552545 TI - Suicide and violence in patients with major psychiatric disorders. AB - The relationship between violence directed at the self and violence directed at others has intrigued psychiatrists for several decades. The goal of this study was to examine the relationship between suicide and violence against others in patients with major psychiatric disorders and to compare psychiatric symptoms associated with suicide in violent and non-violent patients. Subjects included physically assaultive psychiatric inpatients and a nonviolent comparison group. Physical and verbal assaults were recorded prospectively for 4 weeks. History of suicide attempts was obtained through chart review and patient interviews. The Brief Psychiatric Rating Scale was administered at the end of the 4 weeks by raters who were blind to both suicidal and violent behavior. The suicide attempters did not differ from the non-attempters on any measure of violent behavior or hostility. Suicide attempts were not accompanied by different symptoms in violent and non-violent patients; however, violence and suicide attempts were accompanied by dissimilar psychiatric symptoms. The physically assaultive patients presented with more severe positive psychotic symptoms than the non-assaultive patients. The suicide attempters, on the other hand, did not differ from non-attempters on psychotic symptoms, but presented with more severe depression and anxiety. The relationship between these symptoms and suicide attempts was noteworthy in its temporal stability, as most of the patients had attempted suicide many years prior to this study. PMID- 15552547 TI - The recommended dosage range: how is it established and why would it ever be exceeded? PMID- 15552548 TI - A really, really long-term followup. PMID- 15552546 TI - The role of monoamine oxidase inhibitors in current psychiatric practice. AB - The use of monoamine oxidase inhibitors (MAOIs) by psychiatrists has declined over the past several decades with the expansion of psychiatrists' pharmacologic armamentarium. This trend has also been driven by concern about food and drug interactions and side effects, as well as waning physician experience with these medications. Many psychiatrists, in fact, never prescribe MAOIs. Recent research has liberalized the MAOI diet and identified symptom presentations more likely to respond to these medications. Thus, clinicians must continue to familiarize themselves with the properties of and indications for prescribing MAOIs. PMID- 15552549 TI - Organization liability: beyond respondeat superior. PMID- 15552550 TI - TealLock 5.20 security software program for handheld devices. AB - The TealLock has a simple graphic interface, and the program is user-friendly with well thought out options to customize security settings. The program is inexpensive and works seamlessly with the Palm OS platform's built-in basic Security application. The developer offers a 30-day free trial version and there is no downside to trying it to see if it meets your needs. It seems to be an effective security software program for psychiatrists who keep confidential and sensitive patient information on their PDAs. In keeping with HIPAA regulations, the TealLock bolsters security for protected health information stored on PDAs or other handheld devices by providing safeguards that address authentication, access control, encryption, and selected aspects of transmission. PMID- 15552551 TI - Irrational fear of AIDS associated with suicidal behavior. AB - This article concerns patients who have an irrational conviction that they have AIDS, despite medical evidence to the contrary, and who, despite medical reassurance, go on to complete suicide or make serious suicide attempts. Patients with such irrational convictions often develop this symptom complex in the setting of extramarital affairs and subsequent feelings of guilt and shame. Two case reports are presented, one of completed suicide and one of serious attempted suicide. Both patients were on inpatient chemical dependency wards at the time of their suicidal acts. A comparison is made to syphilophobia. The literature on the irrational fear of AIDS and syphilophobia is reviewed. Although such cases are not common and suicidality among such patients is not common, it is helpful for clinicians to be aware of the potential risk for serious suicidal acts in patients who develop this irrational belief system. PMID- 15552552 TI - Legal and ethical challenges in telepsychiatry. AB - Telepsychiatry in the 21st century poses a wide range of legal and ethical challenges. The authors review issues related to licensure, credentialing, privacy, security, confidentiality, informed consent, and professional liability in the use of telepsychiatry services and illustrate the discussion with hypothetical clinical vignettes. It is clear that there will be a need in the immediate future to create legal instruments as well as formal professional ethical guidelines for the practice of telepsychiatry. PMID- 15552553 TI - Development of hip dysplasia in puberty due to delayed ossification of femoral nucleus, growth plate and triradiate cartilage. AB - Besides hip dysplasia diagnosed after birth, there are dysplasias that do not develop until puberty, causing subluxation of the femoral head to occur late in the skeletal growth period. These dysplasias have various causes. In the two siblings described in this study, the boy showed a conspicuous delay in the appearance of the femoral nuclei and the fusion of the triradiate cartilages. The fusion occurred at 16 years of age in the boy and at 13-14 years in the girl. This was preceded by conspicuous structural changes, especially in the posterior triradiate cartilage and acetabular roof but also affecting the lateral growth plate of the femoral neck, which was horizontal over two-thirds of the diameter. We know from animal studies that the growth of the triradiate cartilage increases the diameter of the acetabulum but not the depth. The acetabulum is deepened by pressure from the femoral head. As a result of coxa valga and a prolonged period of acetabular expansion, combined with abnormalities of the superior acetabular rim, the femoral heads in these children finally subluxate. Whenever development of the femoral ossific nucleus is delayed during the first year of life, radiographic follow-ups should be instituted at 8 years of age. PMID- 15552554 TI - Evaluation of the subchondral fracture in predicting the extent of femoral head necrosis in Perthes disease: a prospective study of 92 patients. AB - The aim of this study was to evaluate the subchondral fracture as a predictor for the extent of femoral head necrosis in Perthes disease. Out of 392 patients, 92 (23.5%) had a detectable subchondral fracture at the time of diagnosis. There was concordance between predicted Catterall groups on the basis of the extent of the subchondral fracture and the actual Catterall groups at the time of maximal resorption in 61% of the cases, when assessed by an experienced observer. When using the extent of the subchondral fracture to predict Salter-Thompson groups, this observer obtained 89% concordance with the actual Salter-Thompson groups at the time of maximal resorption. The inter-observer agreement between the experienced and a less experienced observer regarding the presence or absence of a subchondral fracture was moderate (weighted kappa 0.59, 87% agreement). When using the extent of the subchondral fracture as a measure of femoral head involvement (Catterall groups), the inter-observer agreement was moderate (weighted kappa 0.46). Patients with detectable subchondral fracture were significantly older (mean 6.5 years) at the time of diagnosis than those without visible fracture (mean 5.2 years). The delay in diagnosis was significantly shorter in the group with subchondral fracture (mean 3.2 months) than among patients without visible fracture (mean 4.9 months). There was no significant difference with regard to sex, pain level, pain localization, or limping gait between the two groups. We conclude that the subchondral fracture is a relatively rare early sign in Perthes disease. When present, it is a useful sign when assessed by an experienced observer as its extent was in fairly good concordance with the extent of femoral head involvement at the time of maximal resorption. Awareness of this radiographic sign will aid the orthopaedic surgeon to establish diagnosis and, to some degree, to predict prognosis early in the course of the disease. PMID- 15552555 TI - Mechanoreceptor evaluation of hip joint capsule and ligamentum capitis femoris in developmental hip dysplasia: a preliminary study. AB - We aimed to determine if there are mechanoreceptors in the hip joint capsule and ligamentum capitis femoris (LCF) of patients with developmental dysplasia of the hip (DDH). We took capsule and LCF biopsies from 20 hips of 20 patients who were operated on because of DDH. The mean age was 10.2 months (range 6-20 months) at the time of surgery. There were 12 girls and eight boys. Teratologic and secondary hip dislocations were not included in this study. Full thickness, 0.5 x 0.5 cm anterior capsule and LCF portions were taken for biopsy specimen. Specimens were stained with hemotoxylin eosin and examined immunohistochemically using monoclonal antibody against S-100 protein. In both analyses no mechanoreceptor was found in any samples of capsule or LCF. In this preliminary study we could not find mechanoreceptors in the local anterior joint capsule and LCF of the hip in children with DDH. We think that additional studies are necessary in order to understand the mechanoreceptor characteristics of the hip joint in children not only with DDH but also in children with healthy hips. PMID- 15552556 TI - The role of AO external fixation in proximal femoral osteotomies in the pediatric neuromuscular population. AB - Internal fixation in proximal femoral osteotomies using traditional devices may be sub-optimal in children with neuromuscular disorders who have small or osteopenic bone. In this population, between 1988 and 2000, we performed 36 proximal femoral varus osteotomies in 28 patients. These were controlled by the AO external fixator. The average age at surgery was 7 years (range, 2-13 years). A mean varus correction of 34 degrees (range, 15-90 degrees) was obtained. Complications consisted of one superficial pin tract infection, one skin breakdown, and one non-union. Other than the non-union, all osteotomies were stable at the time of the fixator removal. The AO external fixator is an effective alternative in maintaining corrective proximal femoral osteotomies in children with fragile bones. PMID- 15552557 TI - Influence of speed variation and age on the asymmetry of ground reaction forces and stride parameters of normal gait in children. AB - The purpose of this study was to assess the influence of age and speed on the asymmetry of ground reaction forces (GRFs) and spatiotemporal parameters (STPs) during normal gait in 4- to 10-year-old children. Forty-seven children walked at three speeds on a treadmill dynamometer (ADAL; Tecmachine, Andrezieux Boutheon, France). Thirty steps were recorded at each speed for each foot for each child. The GRF and stride parameters were normalized to body weight and to body height, respectively. A left-right symmetry index (SI) was calculated for each parameter. The influence of both age and speed on the different SI was examined with a two way analysis of variance. GRF and STP were asymmetric (SI ranged from +/- 1.92% to +/- 45.05%). The SI of forces Fz1 and Fy1 are negative in children aged 4-6 years, indicating that left Fz1 and Fy1 are higher than right Fz1 and Fy1. The cross effect of age and speed on the asymmetries of gait parameters was not significant There was no significant effect of age on the asymmetries of vertical GRF and STP. The asymmetry of vertical propulsive force (Fz3) alone increased with the speed (P < 0.05). This study showed that the different tasks performed by the lower limbs could be responsible for the asymmetry of GRF and STP in children aged 4-6 years. It also shows that upper and lower limits of normal asymmetry of gait parameters are different. Therefore no single criterion value can be used to assess the symmetry of several gait parameters. The small values of upper and lower limits of symmetry indices of vertical forces and stride duration show that the symmetry indices of these variables are reliable measurements and should thus be used in symmetry analysis of gait in normal and disabled children. PMID- 15552558 TI - Ischemic necrosis following clubfoot surgery: the purple hallux sign. AB - Ischemic necrosis, which develops rarely after clubfoot surgery, may have a vascular etiology, since many idiopathic and neurogenic clubfeet have congenital deficiency of the anterior tibial and dorsalis pedis arteries. Dorsalis pedis deficiency is demonstrated more frequently in those clubfeet showing greater deformity. Substantial hypoplasia of the profunda femoris and posterior and anterior tibial arteries was evident in the affected limb of a patient in this series who underwent postoperative arteriography. Herein, we report massive necrosis in seven limbs of six patients after clubfoot surgery and have combined this series with seven previously published cases. Additional cases support our hypothesis that arterial deficiencies put some postoperative clubfeet at risk of perioperative ischemic necrosis. Necrosis occurs in those regions supplied by the congenitally diminished anterior tibial and dorsalis pedis arteries. Knowing that children with congenital vascular deficiency are at risk for ischemic necrosis, surgeons should be alert to the subtle, early signs of ischemia and be prepared to prevent or ameliorate the consequences of this condition. Since hypoperfusion in these postoperative feet is a surgical emergency, we propose clinical guidelines for treatment for this phenomenon, which we have named the purple hallux sign. PMID- 15552559 TI - Dysplasia epiphysealis hemimelica of the hand: two cases at the proximal interphalangeal joint. AB - We report two cases of dysplasia epiphysealis hemimelica (Trevor's disease) at the PIP level of the finger. In one girl the first radiographic signs were seen at the age of 4 years, and progressive clinodactyly required treatment at the age of 7 years. She underwent an osteotomy. The second girl was seen with a clinodactyly and the accessory growth center was present already at the age of 5 years. The intra-articular exostosis was removed. Both had a complete correlation and a full range of motion. PMID- 15552560 TI - Injuries of the scapholunate ligament in children. AB - Wrist arthroscopy was performed in three children (9, 11 and 12 years old) after persisting pain over the dorsal scapholunate region and revealed intraligamentous tear of the scapholunate ligament in two cases and its avulsion from the scaphoid border in one case. Open surgical repair of all lesions was performed after arthroscopy during the same session. After an average follow-up of 2.4 years all patients were pain free and resumed all sports activities. This study shows that different kind of injuries can occur to the scapholunate ligament in the skeletally immature carpus. Surgical repair leads to good results. PMID- 15552561 TI - Delayed union and non-union of the ulna following intramedullary nailing in children. AB - Most studies report little or no problem with union following intramedullary nailing of fractured forearm bones in children. The bone involved in the occasional delayed union is not mentioned except for one delayed union of the ulna following an open fracture. The present paper specifically highlights problems with union of the ulna following nailing in children. It reports two cases of delayed union and one non-union following nailing of closed fractures of both forearm bones. In all cases, the radius united in good time. We discuss the anatomical reasons and surgical techniques that predispose the ulna bone to delayed union and non-union and recommend surgical precautions to avoid this. PMID- 15552562 TI - Juxtaposition artefact in a forearm radiograph: a cautionary note in the interpretation of a multiply exposed radiographic plate. AB - This case report outlines the risks associated with double plate exposure radiographs in follow up images of a child's forearm fracture. Protocols need to be established between radiology and orthopaedic departments for adequate imaging techniques of children's forearms. PMID- 15552563 TI - Non-adjacent spondylolisthesis in ehlers-danlos syndrome. AB - To report a case of non-adjacent spondylolisthesis in a patient with Ehlers Danlos syndrome. Clinical case analysis of a case of Ehlers-Danlos with non adjacent spondylolisthesis. Review of this case reveals progressive lytic non adjacent spondylolisthesis in a child with Ehlers-Danlos syndrome. There are no other reported cases of Ehlers-Danlos syndrome associated with non-adjacent spondylolisthesis. A review of the pathophysiology of the disease reveals possible mechanisms for this spinal abnormality. PMID- 15552564 TI - Mesomelic dwarfism in pseudoachondroplasia. AB - Pseudoachondroplasia (PSACH) is associated with mutations in the cartilage oligomeric matrix protein (COMP) gene and the clinical characteristics include short stature, deformities of the extremities involving the epiphyses and metaphyses, early onset arthritis, and ligament laxity. PSACH has been considered a rhizomelic form of dwarfism. So far no previous report has described mesomelic shortening of the limbs in PSACH. We reviewed nine patients with a diagnosis of PSACH based on clinical and radiographic examination and mutation analysis of the COMP gene. The mean height in the adults was 116 cm. All patients showed mesomelic dwarfism. The average ratios of radial length to humeral length and tibial length to femoral length were 0.62 and 0.63, respectively. The tibia and the radius showed more severe bony deformity than the femur and humerus. The degree of short stature was related to the site of the mutation in the COMP gene, but there was no correlation between bony deformity and height or gene mutation. PMID- 15552565 TI - Treatment of severe late onset Perthes' disease with soft tissue release and articulated hip distraction. PMID- 15552566 TI - Connecting bar for hip spica reinforcement: does it help? PMID- 15552567 TI - Guidewire damage during connulated screw fixation for slipped capital femoral epiphysis. PMID- 15552568 TI - Treatment of true developmental dysplasia of the hip using Pavlik's method. PMID- 15552569 TI - Hangman's fracture caused by suspected child abuse. PMID- 15552570 TI - [Influence of environmental factors in early ontogenesis on aging and life span]. AB - A possible dependence of aging and life span in man on specific features of early ontogenesis has been analyzed on the basis of published and author's data. It was shown that the life span depended on climatic factors acting during prenatal and/or early postnatal period. When analyzing a sample of 101,634 humans died in Kiev in 1990-2000, a reliable relationship was established between the age of death and month of birth. The lowest and highest death ages were observed in persons born in April-June and in the end of the year, respectively. The lowest and highest average monthly values of death age differed in men and women by 2.6 and 2.3 years, respectively. In persons survived for more than 60 years, a "birthday" effect was demonstrated: dependence of the mortality level on the month of individual annual cycle was the highest in its first and last month. It has been proposed that this effect is related imprinting of the "birth stress" in the structure of biological rhythms, which may lead to periodic changes in viability during the individual annual cycle. The mechanisms underlying long-term effects of stresses in early ontogenesis have been studied on Drosophila melanogaster. X-irradiation of eggs at 0.50 and 0.75 Gy led to an increased level of survival of imago. The described radiation hormesis was accompanied by an increased resistance of DNA to S1-nuclease, which could be due to a long-term activation of the repair system in irradiated insects. The results obtained suggest that long-term changes in the range of gene expression induced by mild stresses in early ontogenesis can be a key mechanism of early "programming" of the potential of longevity and hormesis for life span. PMID- 15552571 TI - [Localization of chromosomal nucleus organizing regions in one-cell mouse embryos and oocytes by fluorescence in situ hybridization]. AB - In mouse zygotes, ribosomal genes (rDNA) are transcriptionally silent and so called "nucleolar precursor bodies" are present instead of typical nucleoli. However, the functional significance of these structures remains obscure. Specifically, it remains unknown whether structural association between the nucleolar precursor bodies and rDNA are maintained when rDNA synthesis is switched off. Here, we studied for the first time the rDNA topology in one-cell mouse embryos and MII oocytes using fluorescence in situ hybridization and mouse rDNA probes. Our data suggest that in the pronuclei of one-cell embryos, rDNAs form rather compact clusters, whose number does not exceed that of nucleolus organizing chromosomes characteristic for the haploid set of mouse chromosomes. In zygotic pronuclei, not all nucleolar precursor bodies are associated with rDNA and not all rDNA repeats are attached to the nucleolar precursor bodies. Altogether, these data favor the idea that spatial interactions of nucleolus organizing chromosomes and nucleolar precursor bodies are not obligatory. We assume that associations between nucleolar precursor bodies and nucleolus organizing chromosomal regions are mediated by centromeric heterochromatin. The total numbers of silver stained nucleolus organizing chromosomes in CBA and C57BL mice are different. rDNA genes are unequally distributed among nucleolus organizing chromosomes and nucleolus organizing regions of sister chromatids. PMID- 15552572 TI - [Influence of nitric oxide synthase inhibitors on embryonic destructions and morphological features in pre- and postimplantation mouse embryos]. AB - Nitric oxide is an important intraovarian regulatory factor. The periimplantation period is a critical phase in mouse development. Although it was shown that nitric oxide plays an essential role during gestation, its role in the preimplantation period is not yet fully clear. We studied the involvement of nitric oxide in developmental competence (embryonic defects and morphology of pre and postimplantation embryos) using nitric oxide synthase inhibitors, which suppress all forms of nitric oxide synthase, and female mice, to which the inhibitors had been administered before their mating with intact males. The level of mortality of pre- and postimplantation embryos in females mated to intact males increased soon after the administration of inhibitors. Studies of the morphology of embryos have shown that there was a delay in embryogenesis at the stages of cleavage and gastrulation. The results obtained suggest that nitric oxide is a potent regulator of embryonic differentiation, specifically in pre- and postimplantation mouse embryos. PMID- 15552573 TI - [Involvement of gap junctions in stimulation of in vitro maturation of the common frog oocytes by low progesterone concentrations]. AB - The inhibitor of gap junctions 18alpha-glycerretinic acid inhibits the maturation of follicle-enclosed common frog oocytes stimulated by low progesterone concentrations. The inhibitory effect of 18alpha-glycerretinic acid does not depend on concentrations within the limits of 5-40 microM. The inhibitory progesterone concentrations differ markedly in different females. The inhibitory effects of actinomycin D (5 microg/ml) and 18alpha-glycerretinic acid (5 microg/ml) were expressed when the same progesterone concentrations were used. When injected in an intact oocyte, Lucifer yellow was transported into the follicle cells, thus suggesting the presence of gap junction between these latter and the oocyte. The data obtained suggest that the previously described transcription-dependent factor formed in the follicle cells under the influence of low progesterone concentrations and stimulating oocyte maturation is transported in the oocyte along gap junctions. PMID- 15552574 TI - [Development of a model for the role of mitogens initiating repair hyperplasia in big salivary glands]. AB - It was shown in repeated experiments on 132 rats that proliferation in the corneal epithelium was nonspecifically enhanced after sialotomy. PMID- 15552575 TI - [Age-related variability of serum protein antigens in sturgeons (Acipenseridae, Acipenseriformes)]. AB - We studied the variability of antigenic properties of proteins in two sturgeon species at different stages of postembryonic development. The deepest changes occurred in individual components of albumins and beta-globulins (transferrins) and were mostly related to an increased proportion of the protein accounting for these antigens. Transformation of the main component of albumins A1 into adult antigens was completed in 5-month old fry. The main component of beta-globulins A (component of transferrins) appeared in the blood flow much later than other proteins and could retain the fry features until the age of 3-4 years. Other antigens belonging to alpha1- and alpha2-globulins and the second component of transferrins were more stable and did not undergo substantial changes. The direction of ontogenetic variability of serum antigens in sturgeon fry did not depend on the habitat of adult fish in fresh or sea water. PMID- 15552576 TI - [Effects of arachidonoyl dopamine, haloperidol, and their mixtures on regeneration of freshwater Hydra attenuata]. AB - Arachidonoyl dopamine and haloperidol, both separately and in different combinations, inhibit regeneration of the gastral and basal regions of hydra. In addition, both substances induce stable anomalies of morphogenesis in the form of outgrowths and additional tentacles in gastral regenerates. In the presence of both substances at different combinations, anomalies either do not appear altogether, or exist for a short time, thus suggesting the normalization of morphogenesis. Possible mechanisms underlying the effects of these substances are discussed. PMID- 15552577 TI - [Distribution and cytogenetic features of triploid male goldfish from Azov basin]. AB - When studying uni-bisexual goldfish (Carassius auratus gibelio) populations in the Azov basin in 1995-2000, we found triploid males, which constituted 2.5%, on average, of the total numbers of studied samples. The areas of nuclei of erythrocytes of triploid males were, on average, 1.35 times those in diploid males. At the same optical density of DNA, the sizes of mature spermatozoon heads in triploid males were, on average, 1.8 times smaller than in diploid males, as follows from the data obtained in 1966. The results of similar studies carried out during the period of natural spawning activity in 1997-1999 suggest that the sizes of spermatozoon heads suggest that the sizes of spermatozoon heads in triploid males were, on the contrary, 1.5 those in diploid males. Triploid males were characterized by mosaicism of spermatozoon size and chromosome mosaicism in somatic cells. Electrophoretic analysis for the locus of transferring confirmed the triploid status of this genetic group. The results of comparative crosses of goldfish with different ploidy suggest a high fertilizing capacity of triploid males, as well as normal viability of their progenies. A distinct positive correlation (r = 0.73) was found between the numbers of triploid females and triploid males in mixed di-triploid populations. No significant correlation was found between males and females within di- or triploid populations. PMID- 15552578 TI - [Influence of combined microinjection of gene engineering construct and endonuclease on preimplantation development of mouse embryos in vitro]. AB - We studied the influence of combined microinjection of a gene engineering construct and site-specific endonuclease Sal in the pronucleus on preimplantation development of (CBA x C57BL)F1 mouse embryos in vitro. The rate of survival of the embryos was estimated according to their capacity to develop until the blastocyst stage and hatch from zona pellucida. The results obtained suggest that the microinjection of exogenous DNA jointly with endonuclease SalI at concentrations from 0.1 to 0.01 U/microl decreased reliably the rate of survival, as compared to the control (p < 0.05 and p < 0.01, respectively). However, a decrease of endonuclease SalI concentration in the injection mixture to 0.01 U/microl enhanced the capacity of mouse embryos to develop until the blastocyst stage and hatch from zona pellucida, as compared to the embryos microinjected with exogenous DNA and endonuclease SalI at a higher concentration. PMID- 15552579 TI - Assessment of softball bat safety performance using mid-compression polyurethane softballs. AB - There is currently much debate about the safety of the sport of softball. Batted ball speed and average pitcher reaction time are factors often used to determine safe performance. Batted-ball speed is shown to be the most important factor to consider when determining safe play. Average pitcher reaction time is explained and directly correlated to batted-ball speed. Eleven aluminum multi-wall, three aluminum single-wall and two composite softball bats were tested with mid compression polyurethane softballs averaging 1721+/-62 N/6.4 mm to represent the relative bat-ball performance for the sport of slowpitch softball. Nine men and six women were chosen for this study out of a test group of over three hundred slowpitch softball players. On average, aluminum bat performance results were within the recommended safety limits established by the national softball associations. However, when composite bats were used, their performance results exceeded the recommended safety limits which can pose a significant safety risk. Using aluminum softball bats, batted-ball speeds ranged from 80 to 145km x h(-1) Using composite softball bats, batted-ball speeds ranged from 146 to 161 km x h( 1). The scientific relevance of this study is to provide performance information that can lead to injury prevention in the sport of softball. PMID- 15552581 TI - Analysis of postural stability in collegiate soccer players before and after an acute bout of heading multiple soccer balls. AB - The aim of our study was to determine if any immediate changes in balance were discernable in college soccer players after a specially designed heading session. Eight male and two female skilled collegiate soccer players had a baseline balance pre-test using the Balance Master, followed by heading 20 balls kicked consecutively by a teammate from the touchline to a point near the goal, which was followed by a post-test using the same testing technique. Paired t-tests were used to compare balance ability between pre- and post-test conditions. There was no difference in balance pre- to post-test (85.3% and 86.2% respectively). No significant difference was noted from pre- to post-test in the mean equilibrium scores for conditions three through six on the Sensory Organization Test, with the exception of condition four, which revealed a significant increase from pre- to post-test. We concluded that an acute session of heading soccer balls may not result in balance changes in collegiate soccer players. PMID- 15552580 TI - Optimizing segmental movement in the jumping header in soccer. AB - This study looks at segmental movements in the jumping header from an optimization viewpoint. Investigations on the header so far have focused on head restriction in the movement but have not clarified how and to what extent body segments influence the performance of the skill. In the present study a biomechanical model was used to analyze the jumping header in simulated competition to give a clear picture of an optimized header. Skilled soccer players headed balls at speeds of 13 m x s(-1) the results indicated that the head moves as a free non-restricted segment in the jumping header and should be allowed to do so, even though much soccer literature says otherwise to prevent injuries. The arm movement showed individual characteristics and gave no general advantages in optimizing ball speed after impact in the header The movement of the legs was, on the other hand, the single most important factor in the skill. Therefore, coaches and players shouldfocus on developing muscle strength in the stomach, back and pelvis and should put no restrictions on head and arm movement to optimize the jumping header. PMID- 15552582 TI - Relationships between rigging set-up, anthropometry, physical capacity, rowing kinematics and rowing performance. AB - The general aim of this study was to examine the relations between rigging set up, anthropometry, physical capacity, rowing kinematics and rowing performance. Fifteen elite single scullers participated in the experiment. Each sculler's preferred rigging set-up was quantified using measurements that included oar length, inboard, span, gearing ratio, swivel-seat height, footstretcher-seat height and distance, and footstretcher angles. Rowing performance was assessed using 2000 m race times from the Australian National Selection trials. Selected anthropometric, physical capacity and kinematic variables were also quantified. Several rigging variables were significantly correlated with each other, and with various anthropometric, physical capacity and kinematic variables. The individual variables that had the highest correlations with race time were 2 km ergometer time (r=0.90), mass (r=-0.87), height (r=-0.86), oar length (r = -0.85) and strength (r = -0.84). Overall results of this study indicated that the fastest rowers tend to be the largest and strongest, and that these larger body dimensions are reflected in the choice of rigging settings. Rigging set-up by itself should not be considered to be a primary determinant of rowing performance, but rather a consequence of faster rowers being larger and stronger and scaling their rigging set-up accordingly. To maximise rowing performance it appears important to tune the rigging of the boat to match the rower's size and strength. PMID- 15552583 TI - Reliability of power output during rowing changes with ergometer type and race distance. AB - Coaches, sport scientists and researchers assess rowing performance on-water and on a variety of ergometers. Ergometers are frequently used because of the easier assessment environment. However, there is limited information on the ability of rowers to reproduce mean power or time-trial time when using different rowing ergometers (Concept II and RowPerfect) or completing tests over different distances (500 m versus 2000 m races). To test the efficacy of an intervention on a rower's ability to produce power, or to monitor that ability, it is essential to determine a reliable rowing performance test. The per cent standard error of measurement in performance (assessed by mean power and time-trial time) of fifteen national standard rowers was determined for five repeated 500 m and two repeated 2000 m races on a Concept II and RowPerfect ergometer. The per cent standard error of measurement (% SEM) in mean power between 5x500m races, regardless of gender, was 2.8% (95% confidence limits (CL)=2.3 to 3.4%) for the Concept II ergometer and 3.3% (95% CL=2.5 to 3.9%) for the RowPerfect ergometer (n = 15). Over 2000 m the per cent standard error of measurement in mean power was 1.3% (95% CL 0.9 to 2.9%) for the Concept II ergometer and 3.3% (95% CL 2.2 to 7.0%) for the RowPerfect ergometer The results highlight an increase in per cent standard error of the mean during performance races of less than 2000m on the Concept II ergometer, and performance races on the RowPerfect ergometer compared with the Concept II ergometer over 500 m and 2000 m. The most appropriate protocol for testing the influence of an intervention on the ability of a rower to produce power would be 2000 m races on a Concept II ergometer. PMID- 15552584 TI - Anthropometric and biomechanical field measures of floor and vault ability in 8 to 14 year old talent-selected gymnasts. AB - The aim of this study was to identify the anthropometric and physical prerequisites for high difficulty floor tumbling and vaulting. Twenty 8-14 year old female talent-selected gymnasts performed handstand push-offs, and single and multiple jumps on a portable Kistler force plate. The force curves were analysed using Kistler and Excel software to obtain peak displacement, peak take-off force, and power The gymnasts were also assessed for sprinting, with and without vaulting, and standing broad jump performances. Video footage from the vault take off was analysed using Video Expert II software to obtain the horizontal and vertical take-off velocities. Each gymnast's best vault starting score, three best floor tumbling skills, and anthropometric characteristics were recorded. Statistical analysis included one-way analysis of variance (ANOVA) to examine the effect of age (8-10 years, 11-12 years, 13-14 years) on the performance measures and linear regression analysis with performance start score for vault or best floor tumbling score as the outcome variable. The best regression model for indicating vaulting talent had, as predictor variables, resultant velocity at take-off from the board, squat jump power, and average power during the last five jumps in the continuous bent-leg jump series. The best regression model for indicating floor tumbling ability had, as predictor variables, age, vault running velocity, and reduced ground contact time in a handstand push-off. PMID- 15552585 TI - Technique factors related to ball release speed and trunk injuries in high performance cricket fast bowlers. AB - In this study we analysed technique, ball speed and trunk injury data collected at the Australian Institute of Sport (AIS) from 42 high performance male fast bowlers over a four year period. We found several notable technique inter relationships, technique and ball speed relationships, and associations between technique and trunk injuries. A more front-on shoulder alignment at back foot contact was significantly related to increased shoulder counter-rotation (p < 0.001). Bowlers who released the ball at greater speeds had an extended front knee, or extended their front knee, during the front foot contact phase (p < 0.05). They also recorded higher braking and vertical impact forces during the front foot contact phase and developed those forces more rapidly (p < or =0.05). A maximum hip-shoulder separation angle occurring later in the delivery stride (p = 0.05) and a larger shoulder rotation to ball release (p = 0.05) were also characteristics of faster bowlers. Bowlers suffering lower back injuries exhibited typical characteristics of the 'mixed' technique. Specifically, the hip to shoulder separation angle at back foot contact was greater in bowlers who reported soft tissue injuries than in non trunk-injured bowlers (p = 0.03), and shoulder counter-rotation was significantly higher in bowlers who reported lumbar spine stress fractures than non trunk-injured bowlers (p = 0.01). The stress fracture group was also characterised by a larger hip angle at front foot contact and ball release, whereas a more flexed front knee at ball release characterised the non trunk-injured group. PMID- 15552586 TI - Immunodetection of a hepatitis C virus (HCV) antigen and Thl/Th2 cytokines in cerebrospinal fluid of meningitis patients. AB - Infection with hepatitis C virus (HCV) has become the most important public health problem in Egypt. HCV infection has been implicated in diseases of the central nervous system. Cerebrospinal fluid (CSF) and serum samples from 91 patients with meningitis (62 males and 29 females, mean age of 37 years) were investigated. Anti-HCV antibodies and HCV antigen were evaluated in patients CSF and serum using enzyme linked immunosorbent assay. The levels (mean +/- SD pg/ml) of Th1 cytokines (IFN-gamma and TNF-alpha) and Th2 interleukines (IL-10 and IL-4) were also determined. The anti-HCV antibodies were detected in high percentages both in CSF samples (71%) and in sera (90%). Also, the HCV antigen was detected in about 60% of tested CSF and serum samples. The levels of IFN-gamma and IL-10 cytokines were significantly higher (P < 0.05) in both serum and CSF of patients positive for HCV antigen than those negative. HCV antigen was detected in the CSF of meningitis patients with a significant upregulation of Th1 and Th2 responses. The high incidence of HCV infection may draw light on the etiological role of HCV in the pathogensis of meningitis diseases in our study group. PMID- 15552587 TI - Development of an enzyme-linked immunoassay for sensitive detection of native and recombinant human interferon-gamma using whole IgG fraction as polyclonal tracer. AB - Monoclonal antibodies (mAbs) and polyclonal antibodies (pAbs) against human interferon gamma (IFN-gamma) were produced and used for development of a sensitive enzyme-linked immunosorbent assay (ELISA) for the detection and quantitation of native and recombinant human IFN-gamma in tissue culture fluid and human sera. The human IFN-gamma ELISA was constructed using mAb CAy-IFNg111 as the capture antibody (Ab) and biotinylated polyclonal mouse immunoglobulin G (IgG) as the tracer Ab. The assay is completed within 4 hr at room temperature (RT). The human IFN-gamma ELISA worked in tissue culture medium and human serum and was capable of detecting both recombinant and native human IFN-gamma. The assay dynamic range extended from 16 to 1000 pg/mL and the sensitivity level was less than 3 pg/mL of human IFN-gamma with averaged intra- and inter-assay variation coefficients less than 8% for both. The results demonstrated that without the need of an antigen-affinity purification, biotinylation of protein G purified pAb, obtained from 1 mL of mouse blood, was sufficient for constructing the tracer reagent for the establishment of a highly sensitive ELISA (40,000 test) for the quantitative detection of native and recombinant human IFN-gamma in culture supernatant and human sera. PMID- 15552588 TI - Development and validation of enzyme-linked immunosorbent assays for quantification of anti-methotrexate IgG and Fab in mouse and rat plasma. AB - This laboratory is investigating the use of anti-methotrexate IgG (AMI) and anti methotrexate Fab fragments (AMF) within an inverse targeting strategy that is designed to enhance the pharmacokinetic selectivity of intraperitoneal (i.p.) chemotherapy. The goal of this study was to develop enzyme-linked immunosorbent assays (ELISAs) to determine concentrations of AMI and AMF in mouse and rat plasma. An antigen-specific ELISA was developed for AMI and AMF in mouse and rat plasma. The assay was validated with respect to precision and accuracy by evaluating the recovery of AMI and AMF from mouse and rat plasma samples. Preliminary pharmacokinetic studies of AMI and AMF were performed in Sprague Dawley rats and Swiss Webster mice. The animals were instrumented with a jugular vein cannula and administered AMI or AMF, 15 mg kg(-1) via the cannula. Plasma samples were taken at various time points and analyzed using the ELISA, and the observed concentration vs. time profiles were subjected to non-compartmental pharmacokinetic analyses. Standard curves for the ELISAs were found to be linear over concentration ranges of 0-250 and 0-350 ng mL(-1) for AMI and AMF, respectively. Intra-assay and inter-assay recovery of AMI and AMF from plasma samples were found to be within 15% of theoretical values. Preliminary pharmacokinetic investigations of AMI allowed estimation of AMI clearance to be 0.017 mL kg(-1) min(-1) in the rat and 0.043 mL kg(-1) min(-1) in the mouse. AMF clearance was estimated to be 0.038 and 1.93 mL kg(-1) min(-1) in the mouse and rat, respectively. In conclusion, ELISAs have been developed and validated for quantitation of AMI and AMF in rat and mouse plasma. The assays will allow further investigations of AMI and AMF pharmacokinetics. PMID- 15552590 TI - The fate of perfluoro-tagged metabolites of L-DOPA in mice brains. AB - A novel compound for use in magnetic resonance (MR) imaging was created by covalently bonding multiple 19F atom tags to L-DOPA. Tagging L-DOPA permits bypassing the rate-limiting factor in the biosynthesis of dopamine (DA), the conversion of tyrosine into L-DOPA. The next step in the biosynthetic pathway, the removal of the carboxyl group on the molecule by the enzyme L-aromatic acid decarboxylase (AADC), happens rapidly after L-DOPA is taken up into neurons. In order to be useful as a tool in MR imaging, the novel compound and/ or its perfluoro-tagged metabolites must accumulate in vesicles in dopaminergic neurons. We administered L-DOPA with a nine 19F atom tag (250 mg/kg) to mice pups, waited 1.5 or 3 hr, and used high pressure liquid chromatography (HPLC) to examine neural tissue samples for tagged L-DOPA and tagged DA. The isomer of L-DOPA with the tag bonded at the 5 position yielded the highest conversion to tagged DA at 1.5 hr after an i.p. injection. This study provides the first direct evidence that L-DOPA, tagged with nine fluorine atoms, is taken up into mammalian brain dopaminergic neurons where it is converted to perfluro-tagged DA. The use of these tagged compounds may make it feasible to investigate the uptake and conversion of important neurotransmitter in vivo with fluorine imaging. PMID- 15552589 TI - Evaluation of a microsphere-based flow cytometric assay for diagnosis of celiac disease. AB - The multiplexed particle-based flow cytometric technology proposes a new approach for the diagnosis of autoimmune diseases combining the advantages of conventional methods with the ability to quantitatively determine multiple autoantibodies in the same sample, simultaneously and rapidly. Recently, a commercial kit (FIDIS Celiac, Biomedical Diagnostics, Mane la Valle, France) was introduced for the simultaneous detection of IgA anti-tissue transglutaminase (anti-tTG), IgG, and IgA anti-gliadin antibodies (AGA). This study was undertaken to evaluate and compare the FIDIS Celiac kit with standardized commercial ELISAs (QUANTA Lite, INOVA Diagnostics Inc., San Diego, CA). A disease group consisted of 21 samples from untreated patients with biopsy confirmed celiac disease (CD), and two control groups of historical sera (207 from regular blood donors and 181 from chronically infected hepatitis patients) were studied. All control sera were negative for IgA anti-endomysial antibodies (EmA) and had an IgA concentration above the lower normal limit. Concerning the reproducibility, intra- and inter assay coefficients of variation (CVs) ranging between 2% and 12%, and between 3% and 21%, respectively, were observed. Regarding the diagnostic quality, each assay was compared to the disease diagnosis using the McNemar test and the kappa (K) parameter, while ROC analysis was applied. Generally, the performance of FIDIS assay was proved almost equally adequate to that of ELISA in the detection of IgA anti-tTG antibodies, IgA and IgG AGA. However, the performance of FIDIS assay was found surmounting that of ELISA among hepatitis patients, possibly due to the avoidance of debris and unbound cross contaminants and, hence, the "noise" of such materials in samples under analysis. Taking our results together with the simplicity and the high throughput of FIDIS assay, its overall performance in the diagnosis of CD seems better than that of ELISA. PMID- 15552591 TI - A new end-point for ELISA titrations. AB - This report describes a new ELISA procedure based on end-point titrations. This end-point ELISA takes advantage of the change of color intensity that occurs when peroxidase-containing wells of an ELISA plate are revealed with diaminobenzidine nickel and further intensification with silver: as antibody concentration and, therefore, peroxidase concentration, decreased, the color became stronger in some wells and, afterwards (i.e., at lower antibody and peroxidase concentrations), the color faded toward clear background. It is proposed that the reciprocal of the sample dilution at which the color intensifies can be used as a measure of the sample antibody content. This report verifies the validity and precision of that procedure. PMID- 15552593 TI - Inter-test comparison between filter paper absorbed blood eluate and serum for malaria serology by enzyme immunoassay: an operational feasibility. AB - Antimalarial IgG and IgM were detected by enzyme immunoassay in finger-stick blood samples collected in capillary tubes and also spotted onto Whatman filter paper. Assay was performed in 92 blood samples obtained from 53 falciparum malaria patients, representing 23 fever cases (malaria negative) and 16 healthy individuals. A simple indirect ELISA was done using Plasmodium falciparum lysate and MSP1(19) peptide as antigens. Total IgG and IgM contents were also estimated in individual sera and filter paper eluate by single radial immunodiffusion (SRID). Assay results of both serum and filter paper eluates were compared. The sensitivity and specificity of the assays for IgG measurement were comparable between serum and filter paper eluates (P < 0.001), whereas, in case of IgM, detection level was poor in filter paper eluates as observed by ELISA and SRID. The filter paper eluates may serve the purpose of antigen-specific IgG detection in seroepidemiological surveys. PMID- 15552592 TI - A laboratory method for purification of major cow's milk allergens. AB - Food allergy is responsible for the most frequent allergic reactions in children under 1 year of age. Diagnostic tests such as skin test or specific IgE assay usually need highly purified preparations of allergenic proteins. The aim of the present study was to purify three main cow's milk proteins: casein, alpha lactalbumin, and beta-lactoglobulin as allergenic extracts for first time in Iran. PMID- 15552594 TI - Combination therapy in clinical and cosmetic dermatology: the marriage of device and drug. AB - The first generations of lasers used in clinical and cosmetic dermatology achieved their effects by means of epidermal and dermal ablation. While effective in removing some of the stigmata of photodamage including pigmentary changes and rhytides, vascular abnormalities associated with such conditions as melasma and rosacea, were not sufficiently effective. The new generation of laser and non laser light devices (eg, intense pulsed light or IPL) offer excellent results in the management of clinical and cosmetic conditions, including significant changes in improvement in vascular conditions such as rosacea and actinic damage and stimulating dermal collagen production, without significant injury to the epidermis. The combination of light therapies and topical agents adds to the efficacy of these procedures, particularly in post-procedural maintenance. Light based therapies have been an important addition to the anti-acne armamentarium as they are effective and do not add to the increasing bacterial resistance problem. PMID- 15552596 TI - Combination therapy for solar lentigines. AB - Solar lentigines are benign, hyperpigmented lesions that present a significant cosmetic nuisance for many middle-aged and elderly patients with chronic accumulated sun exposure. While previous monotherapies designed to lighten these lesions offer relatively modest improvement, there are several new treatment options. Combination topical therapy using 2% mequinol/0.01% tretinoin [Solage Topical Solution] has been shown to markedly reduce lesion darkness with few side effects. Chemical peels can give good results either alone or in combination with topical therapy. Cryotherapy is an effective and inexpensive way of treating solar lentigines while IPL and lasers are more costly treatment options. For patients desiring treatment, optimal cosmetic improvement can be achieved using a combination of topical and procedural therapies. PMID- 15552595 TI - Advances in the topical treatment of acne and rosacea. AB - Acne and rosacea are common skin diseases which may present similarly and both involve inflammation. Both can result in significant cosmetic impairment and lead to quality of life decrements if not optimally treated. The conventional approach for both diseases involves the use of topical therapy to treat inflammatory lesions in combination, when needed, with a systemic or topical antibiotic. An important issue in the management of both diseases at present is the need to reduce antibiotic usage due to the increasing problem of bacterial resistance. One of the emerging treatment paradigms that is becoming increasingly useful as an antibiotic-sparing strategy is the use of procedural therapies in combination with medical management. Such procedural modalities include lasers, intense pulsed light (IPL), and photodynamic therapies (PDT). Topical regimens are used pre-treatment and following physical modalities for maintenance of remission. PMID- 15552597 TI - Utilizing combination therapy to optimize melasma outcomes. AB - Melasma is a chronic and recurrent disorder. It has been underdiagnosed and undertreated due to lack of effective therapies and the perception that it is merely a cosmetic nuisance. Hydroquinone, corticosteroids, licorice extracts and kojic acid have been used as monotherapy to treat melasma. However, the present standard of care in melasma therapy is combination therapy. To date, the most effective treatment is a triple-combination agent that contains hydroquinone 4%, tretinoin 0.05% and fluocinolone acetonide 0.01%. In clinical trials, its use led to complete or near-complete clearing of melasma in 8 weeks. A long-term study demonstrated its continuing efficacy and safety for as long as 360 days. In an examination of quality of life parameters, patients using the triple-combination cream showed significant improvements in self-perception by all 1290 patients. Various combinations of melasma therapy, such as chemical peels, particularly as adjuvants to the triple-combination cream, are discussed. PMID- 15552598 TI - The incidence of skin cancer continues to increase at an alarming pace. PMID- 15552599 TI - Efficacy and irritation in the treatment of actinic keratosis with topical 5 fluorouracil. PMID- 15552600 TI - Practice pearl: gargling with cholinergic ophthalmic drops for treating the oral lesions of pemphigus vulgaris. PMID- 15552601 TI - Computerized image analysis of nails affected by fungal infection: evaluation using digital photographs and manually defined areas. AB - Despite the relevant increase in clinical trials on the efficacy of various systemic and/or topical antifungal agents in onychomycosis therapy, the evaluation of the results is largely subjective. The aim of this study was to set up and ensure an objective, reproducible and reliable method to measure nail plate involvement in onychomycosis. In order to validate a specifically designed software for the computerized image analysis of affected areas of the nail, standardized clinical pictures of onychomycosis were prepared by six different clinicians using a sample of 11 affected nails. Diseased areas and total nail plates were measured both on the clinical pictures and on their drawings traced by the different clinicians on transparent tapes adhering to sample nails. The computerized procedure was undertaken by a trained operator who was not a dermatologist. The variation coefficients of measurements on clinical pictures (automatically detected) and on drawings were compared. In addition, the agreement between automatic evaluation and drawing was evaluated by means of Bland-Altman analysis. To consider the effect of possible variations linked to different operators using the computerized method, 11 clinical pictures (one for each clinical case considered) were selected and submitted to computerized image analysis by six different trained operators. The computerized detection of affected nail areas showed a coefficient of variation (vc=8.5%) lower than that observed on drawings (vc=14.7%). The two methods showed appreciable agreement, as demonstrated by Bland-Altman plot. The coefficient of variation of image analysis conducted by six different operators was very low for the total area calculation (vc=0.9%) and acceptable for pathological area detection (vc=4.8%). Based on the results obtained, we conclude that automatic evaluation is a reliable and helpful method for the measurement of the clinical involvement of the nail plate in onychomycosis and for the evaluation of therapies, since it can increase the objectivity and reproducibility of data. However, in a minority of difficult cases, expert dermatological evaluation is needed. PMID- 15552602 TI - Oral isotretinoin in the treatment and prevention of cutaneous squamous cell carcinoma. AB - The clinical uses of oral isotretinoin (13-cis-retinoic acid) are many. They include treatment of acneiform eruptions, diseases of cornification, inflammatory disorders, and neoplastic processes. Isotretinoin has shown greater efficacy in the chemoprevention of squamous cell carcinoma, rather than in the treatment. High-dose isotretinoin has mainly been utilized in those patients at high-risk for multiple new skin cancers where the risk of morbidity from side effects is overshadowed by the benefit to the patient. PMID- 15552603 TI - Primary prevention efforts for melanoma. PMID- 15552604 TI - The excimer lasers. AB - The excimer lasers are a group of lasers that have found wide application in a variety of medical fields including dermatology, cardiology, ophthalmology, and orthopedics. The word excimer refers to excited dimer. These lasers operate in the ultraviolet range, and examples include the 193 nm argon-fluroide, 248 nm krypton-fluoride, 351 nm xenon-fluoride, and of particular interest to dermatology, the 308 nm xenon-chloride. These lasers utilize a mixture of a noble gas and a halogen as a lasing material. They were first used in medicine for their ability to produce cold tissue ablation, but more recently have been used in dermatology as a method of non-ablative phototherapy. PMID- 15552605 TI - Immunomodulators for skin cancer. AB - Therapeutic interventions to augment tumor antigenicity or increase the host's immune response against cancer cells include recombinant cytokines, immune modulators, vaccination with tumor antigens, T cell-based immunotherapy, and gene therapy. We describe the current role of the immunomodulators (up-regulators of the immune response) in the therapy of skin cancer (non melanoma skin cancer, melanoma, lymphoma, Kaposi sarcoma, and extramammary Paget's disease). PMID- 15552606 TI - Facial enhancement and the European experience with Sculptra (poly-l-lactic acid). AB - The primary reason patients seek aesthetic treatments is to combat the signs of aging. However, the majority of facial treatments and procedures fill specific wrinkles or pull-taught sagging skin, without returning the volume and contours of a youthful face. Injectable poly-L-lactic acid (Sculptra) is a synthetic, biodegradable polymer, popular in Europe for the correction of lipoatrophy. The novel technique and mechanism of action of this product require physicians to adjust their practice of treating a specific line to returning volume to a facial area. Sculptra has been used successfully for the correction of nasolabial folds, mid and lower facial volume loss, jaw line laxity, and other signs of facial aging. Sculptra treatment provides a minimally invasive, effective, and prolonged (18-24 months) facial enhancement correction with a low frequency of side effects and no need for allergy testing. PMID- 15552607 TI - Laser-mediated photodynamic therapy of actinic cheilitis. AB - Actinic cheilitis (AC) is a common precancerous condition for which a safe, effective, rapid, and cosmetically favorable treatment is needed. The objective of this study was to assess the safety and efficacy of the long-pulsed pulsed dye laser (LP PDL) (595 nm) with photodynamic therapy (PDT) for the treatment of AC. This study was designed to be a prospective, proof-of concept pilot study to assess safety and efficacy of LP PDL in conjunction with topical 20% 5 aminolevulinic acid solution PDT for the treatment of AC. Control patients received LP PDL alone. The setting was an outpatient clinical research center. A volunteer sample of 21 patients with biopsy-proven AC was enrolled (age range, 42 86 years; skin types I-III). All patients were refractory to prior therapies. Patients with a history of herpes labialis were pre-treated with famcyclovir. Nineteen patients received one-to-three treatments of topical 20% 5 aminolevulinic acid for 2-3 hours, followed by LP PDL (595 nm) at monthly intervals. Two control patients received one treatment with LP PDL alone. Patients in the ALA-LP PDL group were followed at 1, 2, 3, 6, 9, and 12 months. Clearance of AC was assessed by clinical evaluation. Control patients were followed to the one month interval. We observed none-to-mild pain; slight-to moderate erythema; no crusting, purpura, or scarring; treatment time of less than one minute; and complete resolution of post-operative erythema by day three. Complete clearance was achieved in 13/19 (68%) of patients following a mean of 1.8 treatments (7/13 (37%) after one, 2/13 (11%) after two, and 1/13 (21%) after three treatments). Patients were followed for a mean of 4.1 (range 1-12) months. Among the remaining cases, partial clearing was achieved in two patients, recurrence during the follow-up interval was observed in one patient, and failure to follow-up occurred in three patients. Post-operative impetiginization occurred in three patients with erosive AC, which resolved with dicloxacillin therapy. Among the control patients, no clearing was observed. Treatment of AC using LP PDL (595 nm) at nonpurpuric parameters following topical application of 5 aminolevulinic acid at short incubation times is safe and effective. It may offer the advantages of rapid incubation, treatment, and recovery times, minimal discomfort, excellent cosmetic outcome, and good efficacy rates. Patients with erosive AC should receive antibacterial prophylaxis. Multiple treatments may be required for complete clearing. PMID- 15552608 TI - Prednicarbate (dermatop): a review. AB - Prednicarbate is a non-halogenated, double-ester derivative of prednisolone that has become of interest for the treatment of inflammatory skin diseases, for example atopic dermatitis. In the past, topical steroids have been a popular choice for treating these diseases; however, the potential for significant local and systemic adverse events, such as skin atrophy and hypothalamic-pituitary adrenal (HPA) axis suppression have limited their use. Prednicarbate has been found to have a favorable benefit-risk ratio, with low skin atrophy potential, and a high anti-inflammatory action. Prednicarbate may be particularly advantageous when used intermittently in pediatric and elderly patients. PMID- 15552609 TI - Flexural Bazex syndrome associated with tonsillar adenocarcinoma. AB - Bazex Syndrome (acrokeratosis paraneoplastica) is characterized by a psoriasiform skin eruption that favors acral sites and has been associated with underlying malignancy in all reported cases. Squamous cell carcinoma of the upper third of the respiratory system and upper third of the gastrointestinal tract and squamous cell carcinoma of unknown primary with cervical lymph node metastasis are the most commonly associated neoplasms. Here we report a case of flexural Bazex syndrome with tonsillar adenocarcinoma. PMID- 15552610 TI - Undifferentiated connective tissue disease with various pigmentary patterns: a study of three patients. AB - Undifferentiated connective tissue disease (UCTD) is a condition characterized by the presence of clinical manifestations suggestive of a connective tissue disease and at least one non-organ specific autoantibody. In this report I am presenting three types of cutaneous pigmentary changes are presented in three patients which were the clue to the diagnosis of UCTD. PMID- 15552611 TI - Steroid-free pimecrolimus (Elidel) for monotherapy of lichen planus. AB - Lichen planus (LP) is a disease distinguished by pruitic violaceous planar papules containing reticulated white striae. It can occur just about anywhere, but most commonly occurs in the mouth, genital, and distal extremity areas. The general treatment for LP consists of topical or systemic steroids, although a standard accepted treatment is still to be determined. Pimecrolimus has generated recognition as a topical non-steroidal drug labeled for treatment of atopic dermatitis. The proposed mechanism of action of pimecrolimus is inhibition of cytokine production and proliferation. Cytokine obstruction results in the limitation of T-cell propagation, which is the inciting factor in the pathological process of LP This element may prove be advantageous in the treatment of LP. PMID- 15552613 TI - Acneiform eruption induced by Iressa (gefitinib) tablets used to treat non-small cell lung cancer. AB - Many medications have been reported to induce acneiform eruptions. A relatively new chemotherapy drug, gefitinib (Iressa), approved by the Food and Drug Administration in 2003 for the treatment of advanced-stage non-small cell lung cancer (NSCLC), has been reported to cause acne or an acne-like eruption. We report an Asian female who presented with hundreds of erythematous papules and pustules on her face, chest, and back, all of which appeared after starting gefitinib to treat non-small cell lung cancer. PMID- 15552612 TI - Inflammation of seborrheic keratoses caused by cytarabine: a pseudo sign of Leser Trelat. AB - Cytarabine, used for the treatment of acute myelogenous leukemia, can cause a rare reaction of inflammation of existing seborrheic keratoses. We report a case of cytarabine induced inflammation of seborrheic keratoses mimicking a vesicular eruption in 53-year-old man with acute myelogenous leukemia. PMID- 15552614 TI - Unilateral trichomegaly induced by bimatoprost ophthalmic solution. AB - In this report, we describe the case of an 80-year-old woman with unilateral trichomegaly. PMID- 15552615 TI - The BRAF gene is frequently mutated in malignant melanoma. PMID- 15552616 TI - An interview with the new NJDA president. PMID- 15552617 TI - Modern day Magellan gives hope to thousands. PMID- 15552618 TI - Local anesthesia for N.J. hygienists: a winning proposition. PMID- 15552619 TI - Oral pathology quiz #44. Case number 1. Lichen planus. PMID- 15552620 TI - Practical aspects of bioterrorism for dentistry. Part 1. PMID- 15552621 TI - Vascular endothelial growth factor as a therapeutic target in cancer. AB - PURPOSE: The role of vascular endothelial growth factor (VEGF) in angiogenesis, vascular maintenance, lymphangiogenesis, and immune cell function will be explained. The link between angiogenesis and tumor progression and the rationale for targeting VEGF in the treatment of cancer will be discussed. In addition, the various therapeutic approaches to inhibiting VEGF will be reviewed. SUMMARY: Advances in our understanding of the mechanisms underlying tumor progression have translated into a number of novel, biologically-based therapeutic strategies. One approach stems from evidence suggesting that angiogenesis is required for tumor growth and metastasis. While the vasculature is normally quiescent in adults, bursts of angiogenesis are tightly regulated by dozens of stimulatory and inhibitory factors. VEGF is thought to be the most potent direct-acting stimulatory regulator of angiogenesis. Excessive expression of VEGF is found in many types of cancer, and high levels correlate with increased microvascular density, disease recurrence, and decreased survival. CONCLUSION: Inhibiting VEGF appears to be a valid therapeutic strategy and may prevent the growth of new tumor blood vessels, cause regression of the existing tumor vasculature, enhance the anti-tumor immune response, and normalize the vasculature (thereby improving the delivery of chemotherapy). PMID- 15552622 TI - Update on clinical trials targeting vascular endothelial growth factor in cancer. AB - PURPOSE: The clinical trials of vascular endothelial growth factor (VEGF) inhibitors are discussed. SUMMARY: The recent approval of bevacizumab (a recombinant humanized monoclonal antibody to vascular endothelial growth factor) for the treatment of patients with metastatic colorectal cancer heralds a new era in cancer therapy as bevacizumab is the first dedicated antiangiogenic agent available for routine use. Many other agents designed to target VEGF are in development, including other antibodies targeting the receptor or ligand, soluble receptors, and small-molecule inhibitors of receptor tyrosine kinase function. While some agents are still in the preclinical stages of testing, clinical trials have been done or are ongoing with many VEGF inhibitors. Some of the clinical data are very encouraging, although the development of this class of agents has been hindered in some cases by the lack of dose limiting toxicity and the lack of an accurate understanding of the dose-response relationship. Future efforts will be aimed at elucidating the mechanisms underlying resistance and activity, and optimizing the use of these agents alone and in combination with other modalities in clinical practice. Establishing the relative merits of the different anti-VEGF strategies is another area of intense effort. CONCLUSION: The current data on VEGF inhibitors are encouraging in regard to the treatment of cancer and should continue to be studied in relationship to other antiangiogenic agents. PMID- 15552623 TI - Overview of bevacizumab: a new cancer therapeutic strategy targeting vascular endothelial growth factor. AB - PURPOSE: The pharmacology, pharmacokinetics, preclinical and clinical experience to date, and clinical concerns in monitoring patients receiving bevacizumab, recombinant humanized monoclonal antibody to vascular endothelial growth factor (VEGF), are described. SUMMARY: Preclinical research revealed that bevacizumab specifically inhibits VEGF, has activity in multiple cancer cell lines, and is synergistic with several cancer chemotherapeutic agents. In humans, bevacizumab has a long half-life, allowing intravenous administration once every two to three weeks. Dose-limiting toxicities, the formation of antibodies to bevacizumab, and problems with wound healing after surgery have not been observed in clinical trials. A phase II study of bevacizumab in combination with 5-fluorouracil and leucovorin in patients with metastatic colorectal cancer showed promising results (i.e., therapeutic response rate and disease-free progression of survival), although a clear dose-response relationship was not demonstrated and concerns were raised about the potential for thromboembolic events, bleeding, hypertension, and proteinuria. In a phase III study in patients with refractory metastatic breast cancer, bevacizumab doubled the response rate from capecitabine but it did not affect survival. First-line use of bevacizumab with irinotecan, orouracil, and leucovorin produced significant improvements in response rate, duration of response, and survival in a phase III study of patients with metastatic colorectal cancer. Bevacizumab was associated with hypertension, which was readily managed with mild antihypertensive agents, and possibly with gastrointestinal (GI) perforation, but not with serious bleeding, thromboembolism, or proteinuria. Nevertheless, patients receiving bevacizumab should be monitored for GI perforation, bleeding, thromboembolism, hypertension, and proteinuria, especially if they have a condition that predisposes them to these problems (e.g., a history of unusual bleeding or clotting, hypertension, or proteinuria; use of anticoagulants or other medications that affect clotting or coagulation). CONCLUSION: In clinical trials, bevacizumab has shown promise in promoting synergism with other chemotherapeutic agents in the treatment of various cancers. PMID- 15552624 TI - Congress investigates FDA's handling of antidepressant safety information. PMID- 15552625 TI - MI risk prompts rofecoxib withdrawal. PMID- 15552626 TI - Pharmacists around the globe meet in New Orleans. PMID- 15552627 TI - Pittsburgh Poison Center is terrorism response leader. PMID- 15552628 TI - Use of proton-pump inhibitors at a nonprofit, mixed-model health maintenance organization. PMID- 15552629 TI - Exact volume in intravenous solution bags. PMID- 15552631 TI - Pharmacy residency programs: how to find the one for you. PMID- 15552630 TI - Patent expiration for lansoprazole. PMID- 15552632 TI - Assuming the role of a pharmacy director. PMID- 15552633 TI - The two i's of pharmacy. PMID- 15552634 TI - Pharmacotherapy, pharmacogenomics, and the future of alcohol dependence treatment, part 1. AB - PURPOSE: The neurobiological basis of alcohol dependence, established pharmacotherapies for alcohol dependence, pharmacotherapies under investigation, and obstacles to treatment are discussed. SUMMARY: Alcohol binds to hydrophobic pockets of proteins, changing their three-dimensional structure and their function. Proteins that are particularly sensitive to alcohol include ion channels, neurotransmitter receptors, and enzymes involved in signal transduction. Established pharmacologic treatments, notably disulfiram and naltrexone, combined with behavioral therapies, may reduce the amount of drinking, the risk of relapse, the number of days of drinking, and craving in some alcohol-dependent individuals. For many patients, however, these treatments are not effective. Recent advances in molecular and behavior genetics are guiding the development of new drugs; these efforts seek to identify pharmacologic pathways relevant to alcohol dependence and to more effectively match treatments to individuals according to their genetic characteristics. Efficacy and safety concerns for acamprosate have been satisfied; the drug was recently released for marketing in the United States. Medications such as sertraline, ondansetron, topiramate, and aripiprazole represent novel lines of research and are currently being tested for use in the treatment of alcoholism. Even with more efficacious medications, however, a transformation must occur in how alcoholism treatment is viewed, not only by the public but also by clinicians. CONCLUSION: In addition to existing drug treatments for alcohol dependence, many other medications are under investigation, particularly for specific types of alcoholism. Pharmacogenomics is expected to play an important role in this research effort. PMID- 15552635 TI - Pharmacokinetics and safety of extended-release divalproex sodium tablets: morning versus evening administration. AB - PURPOSE: The pharmacokinetics and safety of 1000-mg extended-release divalproex sodium given once daily either in the morning or evening were compared. METHODS: Healthy volunteers 19-55 years of age were enrolled in this open-label, parallel design study. Subjects were randomized to receive extended-release divalproex either in the morning or in the evening. Blood samples were taken immediately before and at 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 121, 122, 123, 124.5, 126, 127.5, 129, 130.5, 132, 133, 134, 135, 136.5, 138, 139.5, 141, 142.5, and 144 hours after the first dose of each six-day regimen. Plasma samples were assayed for total valproic acid concentrations using gas chromatography. Valproic acid pharmacokinetic values were measured, and the safety of each regimen was evaluated. RESULTS: Mean steady-state valproic acid exposure, maximum concentration, and minimum concentration were 1771 mg x hr/L, 86.9 mg/L, and 55.5 mg/L for the morning dosing and 1728 mg x hr/L, 84.8 mg/L, and 57.4 mg/L for the evening dosing regimens, respectively. Adverse events reported by two or more subjects were abdominal pain and somnolence for the morning dosing regimen and asthenia, headache, and pain for the evening dosing regimen. All adverse events were mild or moderate; none caused subject withdrawal from the study. There were no significant differences in the pharmacokinetic parameters (p > 0.51) and safety between groups. Diurnal variation in plasma valproic acid concentrations was minimal with once-daily administration of extended-release divalproex. CONCLUSION: Evening once-daily administration of extended-release divalproex was not associated with substantial differences in pharmacokinetics or safety compared with morning once-daily administration. PMID- 15552636 TI - Predicted serum valproic acid concentrations in patients missing and replacing a dose of extended-release divalproex sodium. AB - PURPOSE: Computer simulations were used to analyze changes in steady-state total plasma valproic acid concentrations when a patient misses a dose of once-daily extended-release divalproex sodium, replaces it at a later time, and resumes scheduled therapy. METHODS: Valproic acid concentration-time profiles were simulated for 1000 hypothetical patients for each of two missed-dose scenarios using a one-compartment (assumes rapid distribution) population kinetic model with nonlinear protein binding. For each scenario, a lognormal distribution of clearance of unbound valproic acid, volume of distribution of unbound valproic acid, protein-binding values, and albumin concentration was generated. All simulations incorporated 20% interpatient variability and 10% residual error. RESULTS: Our pharmacokinetic simulations predicted that the chance for high plasma valproic acid concentrations resulting in clinical toxicity is low when extended-release divalproex doses are replaced within 12 hours followed by resumption of scheduled administration, perhaps due to the controlled, near zero order absorption characteristics of the extended-release formulation. If a patient misses a dose of extended-release divalproex, it should be replaced as soon as the patient remembers. The next dose should be taken at the regularly scheduled time. A missed dose of extended-release divalproex may be replaced up to 12 hours later without any clinically significant change in plasma valproic acid concentrations in a majority of the patients. CONCLUSION: Simulation of two scenarios suggested that a missed dose of extended-release divalproex sodium may be replaced up to 12 hours later without any clinically significant perturbation in plasma valproic acid concentrations in the majority of adolescent and adult patients with epilepsy. PMID- 15552638 TI - Interdisciplinary education program for nurses and pharmacists. PMID- 15552637 TI - Physical compatibility of pemetrexed disodium with other drugs during simulated Y site administration. AB - PURPOSE: The physical compatibility of pemetrexed disodium with selected other drugs during simulated Y-site injection was studied. METHODS: A 5 mL sample of pemetrexed disodium 20 mg/mL in 0.9% sodium chloride injection was combined with 5 mL of a solution of each of 79 other drugs. The other test drugs included antineoplastics, antiinfectives, and supportive care drugs used undiluted or diluted in 0.9% sodium chloride injection or 5% dextrose injection. Visual examinations were performed with the unaided eye in normal diffuse fluorescent light at intervals up to four hours after mixing. Combinations with no obvious incompatibility were examined further with a high-intensity monodirectional light source to enhance visualization of small particles and low-level turbidity. The combinations were also evaluated with a turbidimeter at one and four hours. All combinations without visual incompatibility were assessed with a particle sizer counter. RESULTS: Of the 79 pemetrexed-secondary drug combinations, 55 were compatible for at least four hours. However,mixture with 24 drugs resulted in precipitation (including microprecipitation) and color change. CONCLUSION: Pemetrexed disodium was incompatible with 24 drugs during simulated Y-site administration and should not be administered with them. PMID- 15552639 TI - Role of the director of pharmacy: the first six months. AB - PURPOSE: Interviewing for a director of pharmacy position is discussed, and recommended activities for a pharmacy director's first six months on the job are described. SUMMARY: A prospective pharmacy director should first target the specific positions available that best fit his or her skills. The individual should tour the hospital and the pharmacy for each position of interest. Interviews should include the pharmacy staff, the nursing staff, the direct report manager, and the medical staff. The hospital's financial condition should be assessed and a postinterview analysis conducted. Once hired, the new director should devote the first three months to an assessment of pharmacy services, staff, and customers; a regulatory and accreditation review; and assessments of organizational leadership, information systems, and clinical services. In addition, key documents, such as hospital policies and procedures, should be reviewed. At the end of the first three months, two documents, a summary of the new director's initial assessments and a chart showing current workflow, should be completed. Tasks for the first six months on the job include devising a work plan to address issues, developing reporting systems and metrics, setting priorities, creating staffing and clinical plans, promoting staff development creating and revising procedures, and evaluating outsourcing needs. The director should take a moment from time to time to celebrate successes with the staff. CONCLUSION: The director of pharmacy has one of the most complex jobs in the hospital. Planning, communicating, networking, writing, listening, leading, and motivating are skills necessary for success. PMID- 15552640 TI - Medication-related services in a Mexican pediatric hospital. PMID- 15552641 TI - The curious adventures of Trichophyton equinum in the realm of molecular biology: a modern fairy tale. AB - Sequence analysis of the highly variable internal transcribed spacer (ITS) regions 1 and 2 of the ribosomal DNA (rDNA) of the nuclear DNA has been used to explore the phylogeny of the dermatophytes. Results have led some investigators to recommend that a number of dermatophyte species be reduced to synonymy with other established species. One such recommendation is that both varieties of the zoophilic dermatophyte Trichophyton equinum be reduced to synonymy with the anthropophilic species T. tonsurans. The morphologies of both species are reviewed, as are their roles in human infection, their physiological characteristics and their respective ecologies. Close examination of these attributes shows clear differences between the varieties of T. equinum and T. tonsurans. The significance of the homogeneity of the ITS sequences of these two dermatophytes is discussed in the context of more recent appraisals of the results of ITS sequencing in other fungi. It is concluded that the results of ITS analysis for both varieties of T. equinum and T. tonsurans are indicative only of common ancestry. Similar arguments could be put forward for rescinding other changes in dermatophyte nomenclature that have been published in recent years. PMID- 15552642 TI - Molecular ecology and pathogenic potential of Fonsecaea species. AB - The genus Fonsecaea is revised on the basis of ribosomal DNA internal transcribed spacer (ITS) sequence data. Two species are recognized, F. pedrosoi and the new defined F. monophora. The distinction between these species does not correspond with the classical distinction of F. pedrosoi and F. compacta. The latter appears to be no more than a morphological variant. Both species recognized in this study are agents of human chromoblastomycosis; however, in F. pedrosoi a strict association with this disease is noted, while F. monophora is a more general opportunist. Subspecific randomly amplified polymorphic DNA (RAPD) typing revealed a high degree of strain diversity, although clonal reproduction is also likely to occur. Most strains with Fonsecaea-like morphology isolated from environments to which symptomatic human patients were exposed were found to be more closely related to species of Cladophialophora than to Fonsecaea. PMID- 15552644 TI - Chemiluminescent visualization of superoxide generated by Candida albicans. AB - The high toxicity of reactive oxygen species (ROS) suggested a possible role in the pathogenicity of human pathogenic fungi. We previously reported a chemiluminescence method for measuring ROS generation in Candida albicans. In the present study, we attempted to visualize the ROS, superoxide anion radical (O2-), generated by paraquat (PQ)-stimulated C. albicans using methyl-Cypridina luciferin analog (MCLA) as a chemiluminescence probe. Colonies of a wild-type C. albicans parent strain and its respiration-deficient mutant grown on agar plates were overlaid with a mixture of PQ and MCLA solutions. MCLA-dependent light emission from the colonies was recorded with a Hamamatsu ultralow-light-imaging apparatus with a CCD camera in a light-tight box. In the wild-type strain, marginal regions of growing colonies were strongly illuminated. The light emission from the colonies was extinguished by superoxide dismutase (SOD), proving that the light emission was strictly due to the superoxide anion. However, colonies of the respiration-deficient mutant poorly generated superoxide. Chemiluminecence measurements by a luminometer showed vigorous superoxide generation by the exponential phase cells of the parent strain but weak generation by the stationary phase cells. In the mutant, superoxide generation was weak compared with the parent strain. These results indicate that expansion of the colonies was due to the actively respiring cells located in the marginal regions. To our knowledge, the present report is the first chemiluminescent visualization of ROS including superoxide generated by C. albicans. PMID- 15552643 TI - Value of an inhalational model of invasive aspergillosis. AB - Animal models of invasive aspergillosis have been used for virulence studies and antifungal efficacy evaluations but results have been inconsistent. In an attempt to reproduce human infection, many Aspergillus animal models have utilized a 'pulmonary route' for delivery of conidia, largely through intranasal instillation. However, several radiolabeled particle studies have shown that aerosol delivery is preferable to intranasal instillation to create a more homogenous delivery to the lungs. We hypothesized that an inhalational model would be more robust for studies of invasive aspergillosis pathogenesis and antifungal therapy. We developed an inhalational model of Aspergillus fumigatus infection using a Hinners inhalation chamber and demonstrated by quantitative polymerase chain reaction that this new inhalational model creates a more homogenous murine pneumonia, facilitating analysis of mutant strains and treatment regimens. PMID- 15552645 TI - PCR identification system for the genus Aspergillus and three major pathogenic species: Aspergillus fumigatus, Aspergillus flavus and Aspergillus niger. AB - A PCR system was developed that allowed recognition of three major pathogenic Aspergillus species, namely A. fumigatus, A. niger and A. flavus, in isolates obtained from clinical specimens. The primer pair for PCR was designed from conserved sequences of internal transcribed spacer 1 (ITS1) ribosomal DNA and its flanking regions. Products 521 bp in size were successfully amplified by PCR from the seven Aspergillus species most frequently encountered as opportunistic pathogens, and the three most commonly significant species were identified by separate PCR reactions or nested PCR based on use of species-specific primers. To our knowledge, this is first report of identification of the second and third most frequent pathogenic Aspergillus species using specific PCR amplification. The PCR based identification system reported here will be a powerful tool to control difficult pulmonary fungal infections and to speed the application of effective treatment. PMID- 15552646 TI - Candida albicans PLD I activity is required for full virulence. AB - Phospholipase D1 (PLD1) mutants of Candida albicans are defective in important in vivo and in vitro virulence factors. PLD1 mutants colonize the murine alimentary tract as well as PLD1 sufficient strains. In comparison to PLD1 sufficient strains, the PLD1 mutants: (i) are unable to survive in internal organs after intravenous challenge; (ii) do not decrease the body weights of mice after oral challenge; and (iii) are not lethal for immunodeficient mice after oral challenge. In vitro, the PLD1 mutants show a drastically reduced capacity to penetrate epithelial monolayers and they fail to develop hyphae when grown on solid Spider medium. The morphogenic switch from yeast to hyphae is controlled by multiple parallel signaling pathways that couple specific stimuli to the regulation of several transcription factors. Our data suggest that PLD1 functions in at least one of these pathways regulating morphogenesis in vitro and that while the mutants are able to form hyphae in vivo, the hyphae are defective in their ability to cause oroesophageal and gastric candidiasis and to kill the C. albicans-colonized mice. PMID- 15552647 TI - Retrospective study of feline and canine cryptococcosis in Australia from 1981 to 2001: 195 cases. AB - A retrospective study of 155 cats and 40 dogs diagnosed with cryptococcosis between 1981 and 2001 was undertaken. Age, sex, breed, clinical findings, feline immunodeficiency virus and feline leukaemia virus status (in cats), species of Cryptococcus causing disease and region of domicile were recorded. Associations between variables were tested. Male and female cats were affected equally. Age ranged from 1 to 16 years, with a preponderance of cats aged between 2 and 3 years. Siamese, Himalayan and Ragdoll breeds were over-represented. Rural cats were more frequently infected with Cryptococcus gattii. Retroviral infection was not identified as a predisposing condition and was not correlated with either species of Cryptococcus or physical findings. Most cats had signs of nasal cavity infection, which was typically localised for a substantial period before invasion of adjacent structures or dissemination. Male and female dogs were affected equally. A marked preponderance of young, large breed dogs was noted. Border Collies, Boxers, Dalmatians, Dobermann Pinschers, Great Danes and German Shepherds were over-represented. Cryptococcus species involved was not affected by place of domicile. Although nasal cavity involvement was important, the canine cohort had a greater propensity to develop secondary central nervous system involvement and disseminated disease than feline cases. There were no clinical findings in either cats or dogs which could be reliably used to distinguish disease caused by Cryptococcus neoformans variety grubii from disease caused by Cryptococcus gattii. Both Cryptococcus species appear to be primary pathogens of cats and dogs, with the upper respiratory tract presumed to be the predominant primary site of inoculation in most but not all cases. PMID- 15552648 TI - Isolation, characterization, and regulation of the Candida albicans ERG27 gene encoding the sterol 3-keto reductase. AB - The Candida albicans ERG27 gene which encodes the 3-keto reductase enzyme required for sterol C-4 demethylation was isolated and found to encode a 349 amino acid protein that is 60% identical at the amino acid level to the Saccharomyces cerevisiae Erg27p. A C. albicans erg27 null was created in a strain containing an integrated ERG27 rescue cassette under the control of the pMAL2 inducible promoter. The C. albicans erg27 strain was able to grow only in the presence of maltose indicating that the ERG27 gene is essential. The C. albicans erg27 null showed complete loss of both 3-keto reductase and oxidosqualene cyclase (Erg7p) activities compromising all sterol synthesis. These results suggest that Erg27p inhibitors might be effective antifungals. To explore ERG27 regulation, an erg11 null strain was generated. C. albicans erg6 and erg24 mutants were also employed along with the inhibitors, itraconazole and zaragozic acid A, to characterize ERG27 expression using Northern analysis. Expression was increased two- to fourfold in erg11, erg6 and erg24 backgrounds. However, itraconazole which targets Erg11p (lanosterol demethylase) increased ERG27 expression 10-fold and zaragozic acid A which targets the Erg9p (squalene synthase) increased ERG27 expression fivefold. The azole and erg11 results support other observations that azoles may affect non-sterol targets. PMID- 15552649 TI - Sino-oral zygomycosis due to Absidia corymbifera in a patient with acute leukemia. AB - Fungi belonging to class Zygomycetes become pathogenic in certain predisposing conditions; principally diabetes mellitus, immunosuppression, trauma or burns. We report a case of a 31-year-old man with acute promyelocytic leukemia who developed infection of the sino-oral cavity, due to Absidia corymbifera during a neutropenic phase following induction chemotherapy. A provisional diagnosis of zygomycosis was made by demonstration of broad aseptate branching filamentous hyphae in the scrapings of the palate, which was subsequently confirmed as A. corymbifera by culture. Surgical debridement could not be done due to the thrombocytopenic status of the patient; instead antifungal therapy with amphotericin B was instituted. However, the patient succumbed to the infection after 15 days of its diagnosis. Although infections with Absidia are infrequent, this case highlights the need for its awareness as a potentially lethal opportunistic fungal infection that can present even with short duration of exposure to the usual risk factors. PMID- 15552650 TI - Candida glabrata sepsis secondary to oral colonization in bone marrow transplantation. AB - Candida glabrata has emerged as a common cause of fungal sepsis in bone marrow transplant patients, particularly those receiving fluconazole prophylaxis. Colonization of the lower GI tract and indwelling catheters have been thought to be the primary sources of systemic infection with Candida. We report on a bone marrow transplant patient who developed Candida glabrata sepsis from pre-existing oral colonization. PMID- 15552651 TI - Line-addressable digital-deflection programmable micromirror array. AB - We report a novel digital-deflection programmable micromirror array driven by micromechanical digital-to-analog converters that eliminates the need for electrical digital-to-analog converters for analog displacement control, thus simplifying the driving circuitry and reducing the overall system cost. Furthermore, owing to the bistable and hysteretic characteristics of parallel plate electrostatic actuators, an array of micromirrors can be controlled by means of row- and column-addressing lines, which drastically reduce the number of routing wires and allow array sizes to increase while they maintain high array quality. PMID- 15552652 TI - Wide-range tuning of polymer microring resonators by the photobleaching of CLD-1 chromophores. AB - We present a simple and effective method for the postfabrication trimming of optical microresonators. We photobleach CLD-1 chromophores to tune the resonance wavelengths of polymer microring resonator optical notch filters. A maximum wavelength shift of -8.73 nm is observed. The resonators are fabricated with a soft-lithography molding technique and have an intrinsic Q value of 2.6 x 10(4) and a finesse of 9.3. The maximum extinction ratio of the resonator filters is 34 dB, indicating that the critical coupling condition has been satisfied. PMID- 15552653 TI - Intensity fluctuations and degree of polarization in three-dimensional thermal light fields. AB - The normalized intensity fluctuations of arbitrary electromagnetic wave fields obeying Gaussian statistics are expressed in terms of the three-dimensional degree of polarization. This general formulation implies an important physical result concerning the polarization of planar fields and the dimensionality of the formalism. The results are expected to be particularly useful in intensity interferometry. PMID- 15552654 TI - High-energy in-fiber pulse amplification for coherent lidar applications. AB - An Er:Yb codoped fiber amplifier chain for the generation of pulses for coherent lidar applications at a wavelength near 1.5 microm is reported. The final 1.8-m long power amplification stage had a 50-microm core diameter and yielded a 23-dB energy gain, resulting in 0.29-mJ, 100-ns pulses at a repetition rate of 4 kHz with no Brillouin scattering and an M2 of 2.1. PMID- 15552655 TI - Spectral polarimetry with a differential group delay bias. AB - We show that the addition of a polarization controller and bias differential group delay to a spectral polarimeter increases the amount of polarization information that can be extracted from a Stokes spectrum measurement. We use this biased spectral polarimetry on 40-Gbit/s signals to measure the polarization mode dispersion of the optical fiber through which they propagate. Our measurements do not require any control of the polarization at the fiber input. Averaging measurements from several settings of the polarization controller yields improved accuracy, and, after data rejection is applied, both the accuracy and the number of valid data points is increased when using several measurements. PMID- 15552656 TI - Germania-glass-core silica-glass-cladding modified chemical-vapor deposition optical fibers: optical losses, photorefractivity, and Raman amplification. AB - Germania-glass-core silica-glass-cladding single-mode fibers (deltan as great as 0.143) with a minimum loss of 20 dB/km at 1.85 microm were fabricated by modified chemical-vapor deposition. The fibers exhibit strong photorefractivity, with type IIa index modulation of 2 x 10(-3). A Raman gain of 300 dB/(kmW) was determined at 1.12 microm. Only 3 m of such fibers is sufficient for constructing the 10-W Raman laser at 1.12 microm with a 13-W pump at 1.07 microm. PMID- 15552657 TI - Characterization of the time dependence of polarization mode dispersion. AB - We elaborate on a recently proposed model of the temporal dependence of polarization mode dispersion in long fiber links. We evaluate the probability that a channel of a wavelength-division multiplexed system never incurs an outage, and the range of frequency over which the parameters that characterize the temporal statistics of the differential group delay are correlated. PMID- 15552658 TI - Adaptive dispersion compensation for remote fiber delivery of near-infrared femtosecond pulses. AB - We report on remote delivery of 25-pJ broadband near-infrared femtosecond light pulses from a Ti:sapphire laser through 150 m of single-mode optical fiber. Pulse distortion caused by dispersion is overcome with precompensation by adaptive pulse shaping techniques, while nonlinearities are mitigated by use of an SF10 glass rod for the final stage of pulse compression. A near-transform-limited pulse duration of 130 fs was measured after the final compression. PMID- 15552659 TI - In-line polarization controller that uses a hollow optical fiber filled with a liquid crystal. AB - We demonstrate an in-line polarization controller based on a hollow optical fiber filled with a nematic liquid crystal by fabricating thin-film electrodes on the cladding of a fiber. The polarization controller consists of three control sections with fixed optic axes, which operate as phase retarders. The phase retardation in each section is controlled by the magnitude of the applied electric field. The full wave retardation voltage of the polarization controller is approximately 85 V. PMID- 15552660 TI - Wide-passband, temperature-insensitive, and compact pi-phase-shifted long-period gratings in endlessly single-mode photonic crystal fiber. AB - We demonstrate what is believed to be the first fabrication of a wide-passband, temperature-insensitive, and compact spectral filter based on a pi-phase-shifted long-period grating in an endless single-mode photonic crystal fiber. By introducing a pi-phase shift in the middle of a 12-period long-period grating, two symmetrical rejection bands at wavelengths of 1252.65 and 1418.7 nm are obtained with isolation higher than 18 dB and a passband bandwidth of 84.15 nm. The pi-phase-shifted long-period gratings are inscribed by the relaxation of mechanical stress with the focused pulses of a CO2 laser and a point-by-point technique without geometric deformation. The length of the spectral filter is approximately 6.6 mm with a sensitivity of 8 pm/ degrees C at a medium temperature range of 23-190 degrees C. PMID- 15552662 TI - Optical biopsy of liver fibrosis by use of multiphoton microscopy. AB - We demonstrate the application of multiphoton microscopy in diagnosing toxin- (CCl4-) induced liver fibrosis in mice. Although hepatocyte autofluorescence does not vary significantly, different degrees of necrosis and stellate cell proliferation at necrotic sites in livers with fibrosis (ex vivo) can be detected easily from multiphoton-induced autofluorescence images by use of 780-nm excitation. Our result suggests that multiphoton microscopy can be developed into an effective technique for the detection and diagnosis of liver fibrosis in vivo. PMID- 15552661 TI - Measuring 0.1-nm motion in 1 ms in an optical microscope with differential back focal-plane detection. AB - Back-focal-plane detection of micrometer-sized beads offers subnanometer resolution for single-molecule, optical trapping experiments. However, laser beam pointing instability and mechanical drift of the microscope limit the resolution of optical-trapping experiments. By combining two infrared lasers with improved differential beam-pointing stability (< or = 0.05 microrad), we simultaneously measure and subtract the motion of the microscope stage, leading to a resolution of <0.1 nm in 1 ms and stability of 0.5 nm over 60 s. Repeated steps of 0.4 nm at 1 Hz are resolved with a signal-to-noise ratio of 25. PMID- 15552663 TI - Anisotropy of light propagation in biological tissue. AB - We investigated the propagation of light in biological tissues that have aligned cylindrical microstructures (e.g., muscle, skin, bone, tooth). Because of pronounced anisotropic light scattering by cylindrical structures (e.g., myofibrils and collagen fibers) the spatially resolved reflectance exhibits a directional dependence that is different close to and far from the incident source. We applied Monte Carlo simulations, using the phase function of an infinitely long cylinder, to explain quantitatively the experimental results. These observations have consequences for noninvasive determination of the optical properties of tissue as well as for the diagnosis of early tissue alterations. PMID- 15552664 TI - Rayleigh scattering cross sections of combustion species at 266, 355, and 532 nm for thermometry applications. AB - Rayleigh scattering cross sections are measured for nine combustion species (Ar, N2, O2, CO2, CO, H2, H2O, CH4, and C3H8) at wavelengths of 266, 355, and 532 nm and at temperatures ranging from 295 to 1525 K. Experimental results show that, as laser wavelengths become shorter, polarization effects become important and the depolarization ratio of the combustion species must be accounted for in the calculation of the Rayleigh scattering cross section. Temperature effects on the scattering cross section are also measured. Only a small temperature dependence is measured for cross sections at 355 nm, resulting in a 2-8% increase in cross section at temperatures of 1500 K. This temperature dependence increases slightly for measurements at 266 nm, resulting in a 5-11% increase in cross sections at temperatures of 1450 K. PMID- 15552665 TI - Experimental observation of true left-handed transmission peaks in metamaterials. AB - We report true left-handed (LH) behavior in a composite metamaterial consisting of a periodically arranged split ring resonator (SRR) and wire structures. We demonstrate the magnetic resonance of the SRR structure by comparing the transmission spectra of SRRs with those of closed SRRs. We have confirmed experimentally that the effective plasma frequency of the LH material composed of SRRs and wires is lower than the plasma frequency of the wires. A well-defined LH transmission band with a peak value of -1.2 dB (-0.3 dB/cm) was obtained. The experimental results agree extremely well with the theoretical calculations. PMID- 15552666 TI - Er:Yb-doped waveguide laser fabricated by femtosecond laser pulses. AB - Laser action is demonstrated in a 20-mm-long waveguide fabricated on an Er:Yb doped phosphate glass by femtosecond laser pulses. An output power of 1.7 mW with approximately 300 mW of pump power coupled into the waveguide is obtained at 1533.5 nm. Waveguides are manufactured with the 520-nm radiation from a frequency doubled, diode-pumped, cavity-dumped Yb:glass laser operating at a 166-KHz repetition rate, with a 300-fs pulse duration. PMID- 15552667 TI - Compact 10-GHz Nd:GdVO4 laser with 0.5-W average output power and low timing jitter. AB - We demonstrate a compact, diode-pumped Nd:GdVO4 laser with a repetition rate of 9.66 GHz and 0.5-W average output power. The laser is passively mode locked with a semiconductor saturable absorber mirror (SESAM), yielding 12-ps-long sech2 shaped pulses. For synchronization of the pulse train to an external reference clock, the SESAM is mounted on a piezoelectric transducer. With an electronic feedback loop of only a few kilohertz loop bandwidth we achieved a rms timing jitter of 146 fs (integrated from 10 Hz to 10 MHz). This is an upper limit because it is mostly limited by the measurement system. The laser setup with a simple linear cavity has a footprint of only 130 mm x 30 mm. PMID- 15552668 TI - Efficient 1521-nm Nd:GdVO4 Raman laser. AB - An efficient compact diode-pumped actively Q-switched Nd:GdVO4 laser at 1521 nm is demonstrated by use of the nonlinear optical process of stimulated Raman scattering. With a 13.6-W diode pump power, greater than 1.18 W of 1521-nm average power at a repetition rate of 20 kHz is generated. At a repetition rate of 5 kHz, the pulse width is shorter than 5 ns and the peak power is higher than 20 kW. PMID- 15552669 TI - Active correction of thermal lensing through external radiative thermal actuation. AB - Absorption of laser beam power in optical elements induces thermal gradients that may cause unwanted phase aberrations. In precision measurement applications, such as laser interferometric gravitational-wave detection, corrective measures that require mechanical contact with or attachments to the optics are precluded by noise considerations. We describe a radiative thermal corrector that can counteract thermal lensing and (or) thermoelastic deformation induced by coating and substrate absorption of collimated Gaussian beams. This radiative system can correct anticipated distortions to a high accuracy, at the cost of an increase in the average temperature of the optic. A quantitative analysis and parameter optimization is supported by results from a simplified proof-of-principle experiment, demonstrating the method's feasibility for our intended application. PMID- 15552670 TI - Diode pumping of a continuous-wave Pr3+-doped LiYF4 laser. AB - We report, for the first time to our knowledge, diode-pumped cw laser oscillation of Pr3+:LiYF4 in the red spectral range. The pump power is provided by a GaN laser diode emitting a maximum output power of 25 mW at a wavelength of approximately 442 nm. The Pr3+ laser emits 1.8 mW of output power at a 639.7-nm wavelength. Threshold pump power and slope efficiency in a nonoptimized setup are determined to be 5.5 mW and 24%, respectively. PMID- 15552671 TI - Low-threshold two-dimensional annular Bragg lasers. AB - Lasing at telecommunication wavelengths from annular resonators employing radial Bragg reflectors is demonstrated at room temperature under pulsed optical pumping. Submilliwatt pump threshold levels are observed for resonators with 0.5 1.5-wavelength-wide defects of radii 7-8 microm. The quality factors of the resonator modal fields are estimated to be of the order of a few thousand. The electromagnetic field is shown to be guided by the defect. Good agreement is found between the measured and the calculated spectra. PMID- 15552672 TI - Enhanced optical nonlinearity near the photonic bandgap edges of a cholesteric liquid crystal. AB - The third-order Kerr nonlinear optical effect of a one-dimensional photonic bandgap structure of a cholesteric liquid crystal is investigated. In a femtosecond nonlinear transmission measurement, nonlinear optical changes in the bandgap edges are observed. From analysis of the dispersion relation, Kerr nonlinear coefficients of nematics, forming the cholesteric liquid crystal, are found to be enhanced by 1-2 orders of magnitude through the photonic bandgap structure. PMID- 15552673 TI - Dependence of parabolic pulse amplification on stimulated Raman scattering and gain bandwidth. AB - An analytical model has been developed and verified by numerical simulations to determine limits induced by stimulated Raman scattering (SRS) on parabolic pulse evolution in high-power, high-energy Yb-fiber amplifiers. Our results show that the maximum achievable parabolic pulse energies are limited by SRS at low amplifier gains and by the finite gain bandwidth at high gains. Therefore, an optimum gain value exists that maximizes the achievable output pulse energy. PMID- 15552674 TI - Efficient generation of high-power quasi-single-cycle terahertz pulses from a single infrared beam in a second-order nonlinear medium. AB - It is shown that the coherence lengths for terahertz (THz) generation based on difference-frequency generation within an ultrafast infrared pulse can be sufficiently long for a wide bandwidth about each phase-matching wavelength owing to a slight dispersion in the THz region. As a result, quasi-single-cycle THz pulses can be efficiently generated. An efficient conversion for the parametric process is made possible not only by use of the wide phase-matching bandwidth but also by optimization of the pulse width for each peak THz frequency. I have investigated the strong-pump regime and found the limits to the conversion efficiencies to generate high peak intensities efficiently for THz waves with which to explore nonlinear regimes of the THz interactions. PMID- 15552675 TI - Diffraction and localization in low-dimensional photonic bandgaps. AB - We show that, in low-dimensional photonic bandgaps, wave diffraction resulting from localization in the translational-invariant directions is strongly influenced by the photonic band structure of the periodic crystal, leading to new kinds of wave localization. In particular, for a periodic layered structure we show that, close to a bandgap edge, diffraction is enhanced, with a transition from a parabolic diffraction curve-typical of isotropic media and supporting Gaussian beams-to hyperbolic or elliptic diffraction curves. In the last two cases localization in the form of stationary X-shaped or sinc-shaped waves is possible. PMID- 15552676 TI - Measurement of temperature profiles on visible light-emitting diodes by use of a nematic liquid crystal and an infrared laser. AB - We present a new technique for measuring the temperature profiles of visible LED chips by use of a nematic liquid crystal with IR laser illumination. The LEDs studied have a multi-quantum-well InGaN/GaN/sapphire structure. New features in this technique are the use of a high-power IR laser beam as the sensing light and the insertion of a color filter in the optical path to block the high-intensity LED light. For the LEDs measured, the conversion efficiency decreases by 70% when the junction temperature rises from 25 to 107 degrees C. This technique is a valuable tool for studying the performance of LEDs as a function of junction temperature. PMID- 15552677 TI - Enhanced single-photon emission from quantum dots in photonic crystal waveguides and nanocavities. AB - A theoretical formalism is presented to investigate enhanced radiative decay of excited dipoles in photonic crystal waveguides and nanocavities with a view to achieving efficient single-photon emission from embedded quantum dots. Surprisingly, large enhancement effects are achievable in both waveguides and nanocavities, and enhanced emission in the waveguide is shown to scale proportionally (inversely) with the photon group index (velocity). Further, a way to include radiative coupling of the quantum dot is shown, and the importance of its inclusion is subsequently demonstrated. PMID- 15552678 TI - Nanostructures, local fields, and enhanced absorption in intense light-matter interaction. AB - Recent literature has reported impressive enhancements in hard-x-ray emission from short-lived solid plasmas by modulation of the interacting surface with nanostructures. We show that the modification of local electric fields near surface structures results in excessive absorption and enhanced x-ray production. A simple model based on local field variations explains the observed x-ray enhancements quantitatively. PMID- 15552679 TI - 11-W average power Ti:sapphire amplifier system using downchirped pulse amplification. AB - We demonstrate a high-power laser system that employs a new scheme in which pulses with negative chirp are amplified and then recompressed by dispersion in a block of transparent material. This scheme has significant advantages for amplification of intermediate energy pulses at high average power, including insensitivity to small misalignments of the pulse compressor, elimination of compressor gratings and their thermal loading issues, low compressor energy and bandwidth throughput losses, and a simplified optical design. Using this scheme, we demonstrate what we believe is the highest-average-power single-stage Ti:sapphire amplifier system with 11-W compressed output. PMID- 15552680 TI - Coherent control of phonon-polaritons in a terahertz resonator fabricated with femtosecond laser machining. AB - Using femtosecond laser machining, we fabricated a terahertz resonant cavity in LiNbO3. Optical pulse sequences with variable repetition rates, generated through a novel pulse-shaping method, are used for characterization of the cavity resonances and for amplification of terahertz phonon-polaritons in the cavity. PMID- 15552681 TI - Optical rectification at metal surfaces. AB - The emission of freely propagating terahertz (THz) radiation coming from optical rectification at metallic surfaces has been detected and characterized for the first time to the authors' knowledge. The observed THz transients are induced through nonlinear electronic processes at gold and silver surfaces on intense pulsed optical photoexcitation and exhibit a peak electric field of as much as 200 V/cm. This finding opens a qualitatively new way to investigate nonlinear phenomena at metal surfaces and also can be exploited for the development of new THz emitters. PMID- 15552682 TI - Plasmas with an index of refraction greater than 1. AB - Over the past decade, x-ray lasers in the wavelength range 14-47 nm have been used for interferometry of plasmas. As in optical interferometry of plasmas, the experimental analysis assumed that the index of refraction is due only to free electrons. This makes the index of refraction less than 1. Recent experiments in A1 plasmas have shown fringe lines bending the wrong way as though the electron density were negative. We show how the bound electrons can dominate the index of refraction in many plasmas and make the index greater than 1 or enhance the index such that one would greatly overestimate the density of the plasma using interferometry. PMID- 15552683 TI - "A moment of grace". St. Joseph Health System looks forward to new life as a public juridic person. PMID- 15552684 TI - Words, actions, beliefs: the mission at work. PMID- 15552685 TI - Integrating spirituality and work. In response to a survey, mission leaders suggest a variety of methods and tactics. PMID- 15552686 TI - Toward workplace spirituality. St. Louis-based Ascension Health is attending to the "spirit in work". PMID- 15552687 TI - Credit where it is due. PMID- 15552688 TI - Patient safety and the ministry. PMID- 15552689 TI - Quality and the "efficacious work of God". The ministry is the sacramental presence of Jesus' touch in today's world. PMID- 15552690 TI - Contract or convenant? The gospel context of Catholic health care should ease the disclosure of medical errors. PMID- 15552691 TI - Medical error: some ethical concerns. The best protection against error remains the physician's traditional sense of professional responsibility. PMID- 15552692 TI - Maine care center reduces use of bed rails. St. Marguerite d'Youville Pavilion launched its initiative in 2003. PMID- 15552693 TI - The "Sentinel Events" study. For our ministry, building a culture of safety should be a leadership responsibility. PMID- 15552695 TI - Diversity in multiinstitutional settings. Interview by Sr. Karin Dufault. PMID- 15552694 TI - "Mercy meds" boosts safety. An initiative at a St. Louis-based system reduces the danger of medication errors. PMID- 15552696 TI - Baptizing" deceased infants? Is there a Catholic ritual that chaplains can perform to relieve grieving parents? PMID- 15552697 TI - Allowing the elderly to age in place. In Portland, OR, a Catholic-sponsored PACE site provides community-based health care services. PMID- 15552698 TI - Enhancing provider quality. PMID- 15552699 TI - Extending the professional memory: four samples of environmental health history. PMID- 15552700 TI - A systems-based food safety evaluation: an experimental approach. AB - Food establishments are complex systems with inputs, subsystems, underlying forces that affect the system, outputs, and feedback. Building on past exploration of the hazard analysis critical control point concept and Ludwig von Bertalanffy General Systems Theory, the National Park Service (NPS) is attempting to translate these ideas into a realistic field assessment of food service establishments and to use information gathered by these methods in efforts to improve food safety. Over the course of the last two years, an experimental systems-based methodology has been drafted, developed, and tested by the NPS Public Health Program. This methodology is described in this paper. PMID- 15552701 TI - Side-by-side comparison of three sampling methods for aerosolized endotoxin in a wastewater treatment facility. AB - Research studies have established the occurrence of adverse health effects in individuals exposed to organic dusts and water aerosols laden with endotoxin. To determine what exposure levels cause these health effects, it is necessary to quantify airborne endotoxin. Several scientific studies have demonstrated that the quantification of detectable endotoxin is affected by differences in sampling media, analytical method, and aerosol composition. The study reported here performed side-by-side endotoxin sampling using a liquid impinger, a glass fiber filter, and a polycarbonate filter in a wastewater treatment plant. Results show levels of detected endotoxin appear to be highest with the impinger. Coefficients of variation calculated for each sampling method show the glass fiber filter having the least variability when sampling was conducted at the highest endotoxin levels. Lastly, a Spearman rank order correlation test identified an apparent correlation between endotoxin levels obtained with the impinger and the glass fiber filter. PMID- 15552702 TI - Detection of latex allergens by immunoelectron microscopy in ambient air (PM10) in Oslo, Norway (1997-2003). AB - The authors collected ambient air along two highways in Oslo to investigate the annual variations in particulate matter (PM10) and the presence of latex as an outdoor allergen. PMI, was monitored for a period of five years, during which time the use of studded winter tires was reduced. The presence of latex and of common aeroallergens was examined directly on the collection filters with immunoelectron microscopy visualized in a scanning electron microscope. The annual variation in PM10 was similar over the five years of sampling, with increased mass concentrations in winter. Statistical analysis indicated no major effect from the change to nonstudded tires. The most important factors influencing the PM10 concentration were meteorological parameters like wind and rain. Immnunolabeling of the filters showed latex as an outdoor allergen that adhered to carbon aggregates from vehicle emission. The results also indicated cross-reactive epitopes among the common allergens investigated, which for sensitized subjects may add to the risk of developing latex allergy. PMID- 15552703 TI - Environmental health professionals and emergency preparedness: Canadian perspectives. PMID- 15552704 TI - Clinical and electrodiagnostic testing of carpal tunnel syndrome: a narrative review. AB - Carpal Tunnel Syndrome (CTS) is a pressure-induced neuropathy that causes sensorimotor disturbances of the median nerve, which impair functional ability. A clear history that elicits relevant personal and work exposures and the nature of symptoms can lead to a high probability of a correct diagnosis. Hand diagrams and diagnostic questionnaires are available to provide structure to this process. A variety of provocative tests have been described and have variable accuracy. The Phalen's wrist flexion and the carpal compression tests have the highest overall accuracy, while Tinel's nerve percussion test is more specific to axonal damage that may occur as a result of moderate to severe CTS. Sensory evaluation of light touch, vibration, or current perception thresholds can detect early sensory changes, whereas 2-point discrimination changes and thenar atrophy indicate loss of nerve fibers occurring with more severe disease. Electrodiagnosis can encompass a variety of tests and is commonly used to assess the presence/severity of neuropathic changes and to preclude alternative diagnoses that overlap with CTS in presentation. The pathophysiologic changes occurring with different stages of nerve compression must be considered when interpreting diagnostic test results and predicting response to physical therapy management. PMID- 15552705 TI - Conservative interventions for carpal tunnel syndrome. AB - The assessment and conservative interventions in patients with carpal tunnel syndrome (CTS) are described in this paper. Information about surgical procedures and postoperative care has also been included. It is difficult to make definitive conclusions about the literature regarding success of treatment for CTS due to variations in outcome measures, severity of CTS, and inconsistencies in duration, dosage, and follow-up time for interventions. Based on what is known to date, this author recommends that patients with mild or moderate CTS be provided with a conservative program of splinting the wrist in neutral for nocturnal wear. In addition, intermittent exercise (nerve-gliding exercises) and activity modification, including avoidance of protracted periods of sustained gripping activities and awkward wrist positions, can be useful. This conservative program may be complemented by pain-relieving modalities during times of activity and supplemental participation in other exercise such as yoga. If symptoms are not relieved to the satisfaction of the patient, or they recur, then it is incumbent upon the therapist to refer the patient to a hand surgeon for injection or possible surgical decompression. PMID- 15552706 TI - Pronator syndrome and other nerve compressions that mimic carpal tunnel syndrome. AB - The purpose of this clinical commentary is to provide a comprehensive review of compressive neuropathies that may mimic carpal tunnel syndrome, provide the clinician with information to differentially diagnose these median nerve compression sites, and provide an evidence-based opinion regarding conservative intervention techniques for the various compression syndromes. While rare in comparison to carpal tunnel syndrome, pronator syndrome and anterior interosseous nerve syndrome are proximal median nerve compressions that may be suspected if a patient with carpal tunnel syndrome fails to respond to conservative or surgical intervention. Differential diagnosis is based largely on the symptoms, patterns of paresthesia, and specific patterns of muscle weakness. Due to the relative rarity of pronator syndrome and anterior interosseous nerve syndrome, few controlled studies exist to determine the most effective treatment techniques. Based on sound anatomical and biomechanical considerations, anecdotal experience, and available research, however, treatment strategies for pronator syndrome and anterior interosseous nerve syndrome compression neuropathies can be divided into 4 major categories: (1) rest/immobilization, (2) modalities, (3) nerve gliding, and (4) nonconservative treatment. PMID- 15552708 TI - Upper extremity work-related musculoskeletal disorders: a treatment perspective. AB - Numerous terms, including repetitive-stress injuries and cumulative-trauma disorders, have been used to describe what is now commonly termed work-related musculoskeletal disorders. The etiology of work-related musculoskeletal disorders is multifactorial and includes physical, individual, and psychosocial factors that contribute to the presenting symptoms. Prolonged positioning away from the ideal posture will affect neural and other soft tissues in the upper extremity. Abnormal postures and positions may result in chronic nerve compression or may shorten muscles and, if the muscle crosses over a nerve, compression may occur. These postures may also contribute to muscle imbalance. A thorough clinical evaluation is necessary to identify all sites of nerve compression, muscle imbalance, and factors affecting patient symptoms so that appropriate intervention can be instituted. Management must include patient education, postural correction, and a specific physical therapy program to address the multiple levels of nerve compression and cervicoscapular muscle imbalance. In addition to physical therapy intervention, behavioral modification at home and at work may be necessary for successful management. PMID- 15552709 TI - Design features of alternative computer keyboards: a review of experimental data. AB - Design of computer keyboards no longer is limited to the flat keyboards that are typically shipped with personal computers. Keyboards now exist that are split into halves and these halves can be slanted away from each other (creating a triangle between the halves), sloped downward toward the visual display terminal, tilted upward like a tent, or simply separated. These design features are intended to alleviate discomfort and possible musculoskeletal disorders that have been suggested to be associated with the extensive use of conventional computer keyboards. The geometry of conventional keyboards requires the wrists to be in 10 degrees to 15 degrees of ulnar deviation and 20 degrees of extension and the forearms to be nearly fully pronated while typing. A review of the available experimental data collected on 10-digit touch typists indicates that (1) keyboards with a slant angle (half of the opening angle) of 10 degrees to 12.5 degrees or keyboards with halves separated to approximately shoulder width are both effective in placing the wrist in near neutral (0 degree) ulnar/radial deviation when typing, (2) wrist extension can be reduced to near neutral (0 degrees) when a keyboard with a negative slope of 7.5 degrees is used, contingent on the wrist rest also sloping with the keyboard, and (3) tilting the keyboard halves 20 degrees to 30 degrees is effective in reducing forearm pronation to approximately 45 degrees. These studies also indicate that experienced 10-digit touch typists readily adapt (within 10 minutes) to these individual alternative keyboard features, and can type with approximately the same speed and accuracy as with the conventional keyboard. While placing the wrist and forearm in a more neutral position could, in theory, reduce the incidence of musculoskeletal disorders, randomized controlled trials are necessary before strong recommendations can be made on the effectiveness of alternative keyboards for the prevention and/or treatment of musculoskeletal disorders. In the absence of these randomized controlled trials, the information in this article provides preliminary guidance to clinicians in their evaluation of computer keyboards and workstations and their recommendations to patients. PMID- 15552707 TI - Work-related musculoskeletal disorders of the hand and wrist: epidemiology, pathophysiology, and sensorimotor changes. AB - The purpose of this commentary is to present recent epidemiological findings regarding work-related musculoskeletal disorders (WMSDs) of the hand and wrist, and to summarize experimental evidence of underlying tissue pathophysiology and sensorimotor changes in WMSDs. Sixty-five percent of the 333 800 newly reported cases of occupational illness in 2001 were attributed to repeated trauma. WMSDs of the hand and wrist are associated with the longest absences from work and are, therefore, associated with greater lost productivity and wages than those of other anatomical regions. Selected epidemiological studies of hand/wrist WMSDs published since 1998 are reviewed and summarized. Results from selected animal studies concerning underlying tissue pathophysiology in response to repetitive movement or tissue loading are reviewed and summarized. To the extent possible, corroborating evidence in human studies for various tissue pathomechanisms suggested in animal models is presented. Repetitive, hand-intensive movements, alone or in combination with other physical, nonphysical, and nonoccupational risk factors, contribute to the development of hand/wrist WMSDs. Possible pathophysiological mechanisms of tissue injury include inflammation followed by repair and/or fibrotic scarring, peripheral nerve injury, and central nervous system reorganization. Clinicians should consider all of these pathomechanisms when examining and treating patients with hand/wrist WMSDs. PMID- 15552710 TI - Clinical and functional differences among patients with idiopathic anaphylaxis. AB - BACKGROUND: There are no studies assesing whether patients with idiopathic anaphylaxis are a heterogenous population OBJECTIVE: A study has been carried out to assess whether clinical and functional differences (mast cell releasability) exist between two sub-types of Idiopathic Anaphylaxis (Generalized Idiopathic Anaphylaxis -IA-G- and Idiopathic Anaphylaxis with Angioedema -IA-A-). METHODS: Patients were selected from the Idiopathic Anaphylaxis (IA) patient population of Hospital General de Albacete (Albacete, Spain) and this data were collected between 1990 and 1995. This series is composed of 81 patients. In the interest of seeing whether an IA classification is warranted between IA-G and IA-A, a logistic regression model was constructed in order to know if differences exist between IA-G and IA-A. To evaluate mast cell releaseability in different groups (IA-G, IA-A, atopic patients, urticaria and healthy subjects) we analysed the log 10 wheal area produced by four consecutive concentrations of codeine (from 90 to 3,3 mg/ ml). In those patients with IA-G, the variable urticaria was controlled, but not in those with IA-A. A parallel line assay was used to study the differences arising among all groups. When the conditions of parallelism and linearity were not fulfilled, a Hotelling's T2 test was performed. RESULTS: In the logistic regression equation total IgE, with an O.R. of 1.006 (95% C.I. 1.001 1.01) favoured the presence of IA-G; whereas the presence of urticaria did not favour the presence of IA-G, with an O.R. of 0.159 (95% C.I. 0.04-0.507). IA-G and IA-A patients showed a higher cutaneous reaction to codeine than atopic patients (p=0.005 and p=0.001 respectively). However, IA-G patients had a lower reaction to codeine than those patients with urticaria (p=0.048). No differences were observed among patients with IA-A and patients with urticaria, as was the case between IA-A and IA-G patients with respect to cutaneous response to codeine. CONCLUSION: Apparently, IA-G patients appear to be closely related to the presence of atopy, while IA-A patients are closely related to the presence of urticaria. Along with other unknown factors, an enhanced mast cell releaseability may explain these episodes of Idiopathic Anaphylaxis among atopic patients. PMID- 15552711 TI - Lysed enterococcus faecalis FK-23 oral administration reveals inverse association between tuberculin responses and clinical manifestations in perennial allergic rhinitis: a pilot study. AB - BACKGROUND: The interest in anti-allergy immunoregulation by lactic acid bacteria (LAB) has been growing over the last few decades. There is evidence to suggest that lysed Enterococcus faecalis FK-23 (LFK), a kind of LAB preparation, could relieve the clinical symptoms of Japanese cedar pollinosis. However, little is known about how LFK plays a role in combating allergy. OBJECTIVE: The aim of this study was to clarify whether improvement of clinical manifestations is associated with enhancement of tuberculin responses in patients with allergic rhinitis treated by LFK. METHODS: One gram of LFK per day was administered orally to fifty perennial allergic rhinitis patients in an open trial that lasted 28 days. Nasal symptoms and sign scores were rated before and after administration of LFK. Tuberculin responses and peripheral blood cells were also measured before and after LFK treatment. RESULTS: Purified protein derivative (PPD) skin test diameter was 2.14 +/- 2.14 mm before LFK administration versus 7.26 +/- 4.81 mm at day 31 (p < 0.01). A significant inverse correlation was recognized between PPD skin test diameters and total nasal scores in the nasal provocation test before and after treatment (r= - 0.600, p < 0.001). Peripheral blood eosinophils were 248 +/- 149 cells/ml before LFK administration and then they significantly decreased to 76 +/- 98 cells/microl (p < 0.01). CONCLUSIONS: These finding may be interpreted as a result of improved clinical symptoms in allergic rhinitis after LFK oral treatment owing to the enhanced host's Th1-type immune responses and supression of the over-expression of Th2-dominated allergic responses. PMID- 15552712 TI - Olerance of a cluster schedule with a house dust mite extract quantified in mass units: multicentre study. AB - The standardisation of allergenic extracts in micrograms of the major allergen has encouraged the search for new treatment schedules, with the purpose of shortening the number of visits and doses required to reach the maintenance dose without eliciting a greater risk of adverse reactions for the patients. With this objective, a prospective multicentre pharmacovigilance study was designed that included 200 patient with allergic rhinoconjunctivitis and/or allergic asthma sensitised to mites (Dermatophagoides pteronyssinu and/or farinae). The dose increment period was carried out using a cluster schedule, where the optimal dose wa reached after 4 visits, administering two doses in each visit. The duration of the study was 5 months and a total o 1902 doses were administered. At the end of the trial, 31 adverse reactions in 23 patients were recorded. Six of these were systemic (0.3% of t administered doses) recorded in 6 patients (3% of the sample). One was an immediate reaction (grade 1) and delayed (4 mild and 1 moderate). Two were asthmatic exacerbations, 2 cutaneous reactions, 1 rhinitis and 1 an unspecific symptom (not IgE-mediated). Two appeared upon administration of the first vial and the remaining 4 after administration of the third cluster. Therefore, the schedule tested presents an adequate tolerance profile, suggesting savings (compared to th conventional schedule of 13 doses per patient) of 1800 visits and 1000 treatment doses in the whole study. PMID- 15552713 TI - Assessment of allergenicity to Mallotus phillipensis pollen in atopic patients in India: a new allergen. AB - OBJECTIVE: Pollen grains of the Euphorbiaceae family are well known causative agents of respiratory allergies in India, European countries and USA. Mallotus phillipensis belongs to the same family and may have some common allergenic properties. It has thus been evaluated for the first time in Indian population for its pollinosis causing properties. METHODS: Pollen antigen of Mallotus phillipensis (MP) was extracted and characterized for its protein components by biochemical methods. Pollinosis potency of crude extract of MP pollen was evaluated by skin prick test on population residing in different parts of India. Specific IgE binding characteristics of the extract were determined by ELISA and Immunoblot. RESULTS: Marked skin reactivity in 5.7% atopic population was recorded and subjects constituting 23.8% of the total patients tested showed skin sensitivity to the MP pollen antigen. Significantly raised specific IgE against MP pollen were recorded in 50% of the skin test positive patients. A number of protein bands were detected in a wide Molecular weight range as well as in acidic pI range, by SDS-PAGE and IEF, respectively. A total 11 protein fractions were detected by the specific IgE antibodies on immunoblotting with patient's sera and were considered allergenic. CONCLUSION: Patients from different geographical regions have shown sensitization to MP pollen antigen. Many proteins have similar molecular weights and pI as other allergenic members of the family (Ricinus communis and Putranjiva roxburghii) found in India, which constitutes a good reason for studying cross reactivity among the members of family Euphorbiaceae, in the future. PMID- 15552714 TI - Analysis of new respiratory allergies in patients monosensitized to airborne allergens in the area north of Milan. AB - BACKGROUND: A recent study carried out in North of Milan, an area that was recently colonized both by birch and ragweed pollen, showed that subjects developing allergies to these "new" allergens were much older than those becoming allergic to "traditional" airborne allergens, which suggests that predisposition to develop respiratory allergies is probably allergen-specific, persists throughout life, and does not occur until the patient encounters the appropriate allergen for a sufficiently long period. OBJECTIVE: The present study aimed to test this hypothesis by following-up a large cohort of patients monosensitized to airborne allergens. METHODS: The prevalence and characteristics of new sensitizations to airborne allergens after >2 years of follow-up were retrospectively assessed in 726 patients monosensitized to grass, pellitory, mite, birch, ragweed, Alternaria, mugwort, or plantain living North of Milan. RESULTS: The overall prevalence of new sensitizations was 35% (256/726), with marked differences between the subgroups ranging from 11% in Alternaria-allergic subjects to 46% in grass-allergic subjects. Patients allergic to grass, birch, or pellitory pollen showed a significantly higher propensity to become sensitized to a second airborne allergen than subjects allergic to ragweed, Alternaria, and mite. Overall, the "new" allergens (birch and ragweed) caused 228/256 (89%) new sensitizations detected in the whole study group. Ragweed pollen induced 90% (38/42) of new sensitizations in birch pollen allergic patients, and birch pollen induced 80% (8/10) of new sensitizations in ragweed allergic patients. No difference in age at the first visit or in duration of the follow-up between patients developing and not developing new respiratory allergies was observed. CONCLUSION: This study shows that: 1) predisposition to develop respiratory allergies is allergen-specific and persists throughout life; 2) proneness to become allergic to certain airborne allergens might be associated with primary sensitization to specific airborne allergens; 3) in monosensitized adults, sensitization to another airborne allergen that has been always present in that particular geographical area is unlikely. PMID- 15552715 TI - Epidemiology of urticaria in Spain. AB - BACKGROUND: In spite of the frequency of chronic urticaria there are very few epidemiological studies of its prevalence and distribution. OBJECTIVE: We wanted to approach the real prevalence of chronic urticaria in a population-based study and to depict demographic distribution and personal perception of the disease. We also wanted to describe the frequency of acute urticaria episodes in the population studied. METHODS: We conducted a population-based study among adults in Spain. We questioned 5003 individuals after calculating a sample size for a maximum variability (conservative approach p=q=0.5). RESULTS: We found a 0.6% (95% CI: 0.4-0.8) prevalence of chronic urticaria. The prevalence is significantly higher in women than in men with a OR=3.82 (95%CI 1.56-9.37). Chronic urticaria is a self-limited disease, yet in 8.7% of cases chronic urticaria lasts from one to 5 years and in 11.3%, for more than 5 years. The average age of onset is 40 years. CONCLUSIONS: We offer large epidemiology study data on the prevalence of chronic urticaria. The prevalence of chronic urticaria has not yet been defined in an adult population-based study. With this work we offer such data to describe the prevalence and features of this disease. PMID- 15552716 TI - Thirty cases of bronchial asthma associated with exposure to pet hamsters. AB - BACKGROUND: The identification, isolation, and elimination of allergen(s) causing bronchial asthma are the most efficient form of treatment. The pet industry has diversified recently, increasing the risk of exposure of pet owners to many unknown antigens. We clinically studied the characteristics of asthma associated with exposure to pet hamsters. METHODS: The study group comprised 30 adults in whom the onset, recurrence, or exacerbation of asthma was triggered by contact with pet hamsters. Clinical characteristics such as sex, age, period required for symptom onset, species of hamster, treatment and disease course, smoking status, and hamster-specific IgE antibodies in serum were studied. RESULTS: The male: female ratio of the study group was 1:1.3, and mean age was 37.7 years. Patients with no previous history of asthma initially presented with cough, progressing to episodes of asthma. Asthmatic symptoms were associated with hamster contact and ranged in severity from mild to severe. Three patients required hospital admission for treatment. The mean period from the start of hamster exposure to the onset of asthmatic episodes was 15.7 months. Dwarf hamsters were responsible for most cases. The CAP-RAST score for hamster-specific IgE antibodies was 1 to 4 in 22 patients and 0 in 8 patients. Eight patients with a score of 1 or higher for hamster-specific IgE antibodies had a CAP-RAST score of 0 for mite antigen. In these patients, terminating hamster contact resulted in a rapid improvement in symptoms, with no need for further treatment. Twenty-three of the 30 subjects (76.7%) were smokers. CONCLUSION: Exposure to pet hamsters is an important risk factor for the onset, recurrence, or exacerbation of asthma. Smoking may also increase the risk of asthmatic symptoms in patients exposed to hamsters. PMID- 15552717 TI - Allergenic fungi spore records (15 years) and sensitization in patients with respiratory allergy in Thessaloniki-Greece. AB - BACKGROUND: The prevalence of respiratory allergy to fungi spores (FS) is not precisely known but is estimated at 20-30% of atopic patients. There are no aerobiological records, necessary for respiratory allergy diagnosis and treatment, in Thessaloniki and generally in Greece. AIM: Creation of a database on FS circulation and investigation of skin sensitivity (SS) of asthmatics by using skin prick tests (SPT). MATERIAL AND METHODS: Daily records and identification of 15 airborne FS species were conducted, using a Burkard trap during 1987-2001. SS to 5 most common FS extracts was investigated, by using SPT in a total of 1311 asthmatics with atopy, submitted to the Out-Patient Clinic of Asthma (Pulmonary Dept, Aristotle Univ, Thessaloniki) in 1990-2001. RESULTS: The FS recorded in the 15-year period were as follows: Cladosporium spp. (72.2%), Alternaria spp. (9.8%), Ustilago spp. (8.1%), Ascospores (2.7%), Agrocybe spp. (1.5%), Helminthosporium spp. (1.4%), Leptosphaeria spp (1.2%), Agrogybe spp. (1.1%), whereas the species Botrytis, Stemphylium, Pleospora, Nigrospora, Epicoccum. Fusarium, Torula and Phoma presented concentrations <1%. The highest numbers of airborne FS were recorded during summer. Positive skin reaction to FS was observed in 421 (32%) patients of the 1311 asthmatics. Positive skin reaction to Altemaria species was observed in 177 patients (13.5%), in 98 (7.4%) to Cladosporium, 65 (5%) to Aspergillus, 45 (3.4%) to Fusarium and 36 (2.7%) to Rhizopus. FS sensitivity is much more frequent in younger men. CONCLUSIONS: For the first time in Thessaloniki-Greece, 15 allergenic FS species circulation has been recorded for the last 15 years. SS was more frequently detected for the species of Alternaria, Cladosporium and Aspergillus. PMID- 15552718 TI - Mite allergen levels and acarologic analysis in house dust samples in Uberaba, Brazil. AB - Mite allergen exposure has been widely related to sensitization and development of allergic diseases. This study intended to evaluate the degree of allergen exposure in Uberaba, Brazil, through the measurements of Der f 1 and Der p 1 allergen levels associated with the acarologic analysis in house dust samples. A total of 240 dust samples were collected from 60 houses through vacuuming sofas and bedding, during the months of March and July 2000. Indoor temperature and relative humidity were also measured. Mites were counted and identified under light microscopy and allergen levels were measured by two-site monoclonal antibody ELISAs. The major mite family was Pyroglyphidae (39.4%), having D. pteronyssinus as the most frequent species (15.6%), followed by D. farinae (12.3%) and E. maynei (7.9%). The family Glycyphagidae was less commonly found (4.8%), with Blomia tropicalis (4.4%) as its majoritary member. The highest levels of Der f 1 and Der p 1 allergens were found in bedding samples in March (31.7 and 0.9 microg/g of dust, respectively), with Der f 1 levels significantly higher than Der p 1 (p < 0.0001). There was a significant positive correlation between the mite number and allergen levels. These results indicate that Dermatophagoides sp are the most frequent mites in our region followed by E. maynei. Therefore, the knowledge of the local mite fauna would improve the means of investigating the association between allergen exposure and sensitization, allowing the addition of new mite extracts in diagnostic tests. PMID- 15552719 TI - Airborne plane-tree (Platanus hispanica) pollen distribution in the city of Cordoba, South-western Spain, and possible implications on pollen allergy. AB - Plane-trees are widely grown in Spain as ornaments. This taxon releases large amounts of pollen into the air and this pollen type seems to have proved implications on pollinosis. The aim of this study was to analyse airborne Platanus pollen content in the air of C6rdoba (South-western Spain) over the last 10 years, and its distribution in different areas of the city, as well as to consider possible implications regarding pollen allergies. Results revealed that Platanus pollen concentrations have been at their highest over the last 5 years. Within the city, the lowest pollen concentrations were in Northern areas of the city. Nevertheless, concentrations high enough to be of risk are reached over the whole city. All the patients studied presented symptoms during the Platanus pollen season. We can therefore conclude that plane pollen presents a risk for the whole city although symptoms are more intense and persistent in districts with a higher number of plane-trees. Platanus allergy is a particular problem for city-dwellers, where these trees are commonly used as ornamentals. PMID- 15552720 TI - Airborne allergy to sunflower seed. AB - BACKGROUND: There is increasing evidence that bird fanciers may develop airborne allergies to unusual allergens. OBJECTIVE: To detect the allergen source in a bird fancier with a history of asthma associated with bird cage cleaning activities and with contact with a Brazil parrot. METHODS: SPT with a large series of both airborne and food allergens were carried out. IgE reactivity to allergens causing wheal and flare reactions was confirmed by in-vitro investigations including ELISA/ELISA inhibition and immunoblot analysis. RESULTS: Strong skin reactivity to sunflower seed was observed. Immunoblot analysis showed IgE reactivity to low m.w. proteins, most probably 2S albumin, and ELISA inhibition studies showed the absence of cross-reactivity to mustard. CONCLUSION: Sunflower seed dust may sensitize patients via the respiratory tract. Differently from previously reported cases of sunflower seed allergy, no cross-reactivity to 2S albumin from botanically unrelated seeds was found. PMID- 15552721 TI - Topical dexketoprofen as a cause of photocontact dermatitis. AB - We reported on the case of a patient who developed a cutaneous eruption in a photoexposed area 1 week after a continous topical treatment with dexketoprofen (Enangel). Photopatch tests were positive for dexketoprofen, ketoprofen and piketoprofen and patch test was positive for piketoprofen. Control photopatch testing with dexketoprofen in 15 healthy volunteers was negative. Dexketoprofen, ketoprofen and piketoprofen are non-steroidal anti-inflamatory drugs (arylpropionic acid derivatives) often used as topical anti-inflammatory agents. It appears that the benzophenone moiety of their chemical structure is the cause of their photosensitivity and cross-photoreaction. PMID- 15552722 TI - A rare case of intravascular coagulation after honey bee sting. AB - A patient presented with coagulation problems a few days after honeybee sting. The purpuric skin changes developed on the legs and buttocks. She manifested signs of hypotension with disturbance of consciousness. Allergen-specific IgE serum levels against honey bee venom antigens reached >17.5 kU/l. The platelet count was 33,000/ml . The prothrombin index decreased to 28%, prothrombine time was prolonged to 34". Fibrin degradation products were present in serum. After 10 day treatment the girl improved, but necrotic skin changes required further plastic surgery. Honeybee sting problems should be taken into account as a cause of coagulation problems. PMID- 15552723 TI - A new theoretical foundation for the polarized-oriented multilayer theory of cell water and for inanimate systems demonstrating long-range dynamic structuring of water molecules. AB - Over the centuries, a vast amount of evidence has been gathering that layers of water sometimes measuring tens of thousands of water molecules thick exhibit altered properties in consequence of exposure to some solid surfaces. Yet, a clear cut theory based on the laws of physics that would predict this kind of long range dynamic ordering of water molecules has been long missing. It is thus with great joy that I announce that a new theory has been developed, which offers theoretical confirmation of the phenomena of long-range dynamic structuring of water by appropriate solid surfaces and which gives clear cut quantitative answers to some key questions about the phenomenon. Thus, for example, under an ideal condition, an idealized checkerboard of alternatingly positively-, and negatively-charged sites of the correct size and distribution could polarize and orient deep layers of water molecules ad infinitum. Based on the quantitative data thus obtained and a relevant simple statistical mechanical law, the new theory predicts that a thin layer of water held between two juxtaposed ideal or near-ideal NP surfaces will not freeze at any (attainable) temperature. On the other hand, water polarized and oriented by an ideal or near-ideal NP-NP system may also not evaporate at temperature hundreds of degrees higher than the normal boiling temperature of water. Both predictions have been confirmed (retroactively) by experimental observations made in the past, accidentally or by design. In a following paper, I will demonstrate that the conclusion reached from the study of the two-dimensional NP surface can be smoothly passed on to the living cells. In the living cell, only one-dimensional linear chains of fully extended protein chains exist. Nonetheless, by proper orientation and distribution, they can achieve similar though less intense water polarization orientation--as experimentally demonstrated worldwide during the 40 years past. PMID- 15552724 TI - Cepharanthine, an anti-inflammatory drug, suppresses mitochondrial membrane permeability transition. AB - Cepharanthine (CEP), a biscocrourine alkaloid, has been widely used in Japan for the treatment of several disorders. Furthermore, accumulated evidence shows that CEP protects against some cell death systems but not others. Recently, it was found that mitochondria play an important role in a mechanism of apoptosis involving membrane permeability transition (MPT). Although CEP stabilizes the mitochondrial membrane structure and protects some functions of mitochondria from damage, the mechanism of action of CEP on MPT remains obscure. In this study, therefore, we examined the effect of CEP on Ca2+- and Fe2+/ADP-induced MPT of isolated mitochondria. CEP inhibited Ca2+-induced swelling, depolarization, Cyt.c release, and the release of Ca2+ in a concentration dependent manner. CEP also inhibited Ca2+-induced generation of reactive oxygen species and Fe/ADP-induced swelling and lipid peroxidation. Furthermore, CEP suppressed Ca2+-induced thiol modification of adenine nucleotide transloase (ANT). These results suggested that CEP suppressed MPT by a decrease in affinity of cyclophilin D for ANT. From these results it was concluded that the suppression of MPT by CEP might be due to its inhibitory action on Ca2+ release and antioxidant activity and that CEP might suppress the mechanism of apoptotic cell death when directly interacted with mitochondria in cells. PMID- 15552725 TI - MR imaging of vertebral metastases at 0.2 Tesla: clinical evaluation of T1 weighted opposed-phase gradient-echo imaging. AB - The purpose of this study was to evaluate clinically T1-weighted spin-echo imaging (T1-SE) and T1-weighted opposed-phase gradient-echo (T1-opposed GRE) in medical examinations for vertebral metastases using 0.2 Tesla MRI. Twenty-one patients (9 males and 12 females, 105 vertebrae) with non-neoplastic lesions were assigned to the normal group (Group N), whereas 27 patients (16 males and 11 females, 133 vertebrae) with malignant metastatic vertebral lesions were assigned to the metastatic group (Group M). Using quantitative analysis, the contrast to noise ratio (CNR) for bone marrow to muscle in the region of interest (ROI) defined by the operator were determined with both groups, whereas the CNR for lesion to bone marrow was determined with Group M. In the subjective analysis, 275 vertebrae of 27 patients in Group M were evaluated. There were significant differences in the Group M between CNR values, T1-SE and T1-opposed GRE. The evaluation by T1-SE image alone was rated as 85.0% for sensitivity, 99.3% for specificity, respectively. On the other hand, evaluation rate only by T1-opposed GRE was 98.5% for sensitivity, 82.4% for specificity, respectively. T1-opposed GRE image is an effective tool for 0.2 Tesla MRI to examine metastatic bone marrow tumors. These findings indicate the necessity of both T1-SE and T1-opposed GRE in diagnostic imaging of vertebral metastases using 0.2 Tesla MRI. PMID- 15552726 TI - Photo-induced arousal response by hornets. AB - When an Oriental hornet Vespa orientalis is subjected to ether anesthesia and then exposed to ultraviolet A light (UVAL) (at a wavelength of 366 nm), it commences showing signs of awakening by starting to move its limbs. While in the process of waking the voltage on its body surface surges sharply from 17-180 mV (median = 71.0) to a level of 93-570 mV (median = 327.5). This elevated level is maintained for several minutes but subsequently drops sharply to starting level. The increase in voltage is throughout accompanied by fluttering of the wings and movements of the legs, as well as attempts to extricate itself from the bindings to the electrodes. These movements by the awakening hornet persist for several minutes even after the irradiation source is turned off but shortly after the switch-off the hornet lapses into sleep again. The described scenario is generally similar in worker, queen and drone hornets, and may even occur in decapitated specimens. The same type of awakening can be repeated in the same fashion after a while, but then the increase in voltage will be smaller than the first time. Continuous UV irradiation of an anesthetized hornet results in a generalized and protracted awakening which, however, is significantly shorter than in a hornet left anesthetized in the dark. PMID- 15552727 TI - [Effects of "host factor" bile on adaptability and virulence of Vibrios, foodborne potential pathogenic agents]. AB - In order to improve the knowledge of host/pathogenic agent interaction and to obtain a more careful estimation of risk related to ingestion of food contaminated by Vibrio spp., the effects of bile extracts have been studied. The growth of one V. fluvialis, two V. alginolyticus, and three V. parahaemolyticus strains, isolated from mollusks and crustaceans, has been determined to evaluate their adaptability to intestinal environment. Moreover, the expression of virulence factors responsible for the colonization, as bacterial "swarming mobility", biofilm production, adherence on epithelial cells and hydrophobicity, has been evaluated. Using a bile concentration of 1.5%, all examined strains showed a constant inhibitory effect, quite moderate in the first growth phases. Bile increased the "swarming mobility" and biofilm production; also the adherence was favored, but only after adaptation and during the early logarithmic phase. The decreased hydrophobicity could explain the reduction of adherence during the stationary phase. Studying the phenotypic expression of virulence factors in "minor vibrios" in the presence of bile, it was possible to extend the knowledge about their pathogenetic mechanisms owing to the ingestion of contaminated food. That permits a more careful estimation of risk related to the contamination, considering the high frequency of isolation of these species in some seafood. PMID- 15552728 TI - [Measles in the Local Health Unit Naples 4: vaccination coverage, epidemic 2001 2002 and prevention strategies]. AB - The authors have analysed vaccination coverage for measles. The active offer of the vaccine brought about an increase of the vaccination coverage but participation to vaccination proved unsatisfactory between 1995 and 2000. The measles epidemic reported 1367 cases, of which 125 were hospitalised (3 cases presented complications). The target for the next years is to reach a vaccination coverage of 95% or more and recuperate the non vaccinated, starting with the 1990 birth cohort. PMID- 15552729 TI - [Health-care workers vaccination coverage: such a long way to run]. AB - The poor vaccination coverage among health-care workers in a hospital, resulted from a self-administered questionnaire, showed the necessity to improve educational efforts and to promote vaccination campaign. PMID- 15552730 TI - [Trihalomethanes and chlorites in finished drinking water in Sardinia (Italy) and possible health effects]. AB - A five years monitoring was carried out in central and southern Sardinia (Italy) to assess the levels of two disinfection by-products (DBPs)--total thrialomethanes (TTHMs) and chlorites--in drinking water Between 1997 and 2002, about 1900 drinking water samples were analysed for both of them. The results showed that TTHMs exceeded very often the maximum admissible concentration. Chlorite concentration was found rising in all the distribution networks since the utilization of chlorine dioxide as disinfectant in 1999, exceeding the maximum contaminant level goals suggested by the World Health Organization. These results are relevant for epidemiological studies on health effects from DBPs exposure. PMID- 15552731 TI - [The prevalence survey of nosocomial infections: a very informative tool in a big hospital setting]. AB - In November 2001 a one-day prevalence survey of nosocomial infections (NI) was conducted in surgical and intensive care units at two hospitals in Rome. The main goal was to quantify the occurrence of NI and of some patient-care practices, such as antibiotic use. The following NI were studied: pneumonia, bloodstream infections, urinary tract infections, surgical site infections (SSI), central line-related local infections. A total of 517 patients were included. The prevalence of the above mentioned NI was 6.6%. SSI were the most frequent NI (50%), with a prevalence of 6.5% in patients who underwent surgery; the SSI prevalence was 4.7% in clean surgical procedures (SP) and 5.6% in clean contaminated SP. In 29 NI, 33 micro-organisms were isolated; the most frequent (24.3%) was Staphylococcus aureus, which was methicillin-resistant (MRSA) in 75% of the strains. On the day of the study, 41.8% of the patients were treated with antimicrobials, mainly aminopenicillins and third generation cephalosporins. In 31.8% of cases antibiotics were given for surgical prophylaxis; patients received prophylactic antibiotics more than 2 hours prior the initial incision in almost 40% of clean SP and 60% of clean-contaminated SP. In our hospitals prevalence survey was a very useful tool to identify the following priorities: SSI prospective surveillance in clean and clean-contaminated SP, control of MRSA selection and transmission, guidelines for antimicrobial prophylaxis in surgery. PMID- 15552732 TI - [Trends in gastric cancer incidence, mortality and survival in the Umbria region of Italy. 1978-82 and 1994-99]. AB - Gastric cancer, despite the decrease observed in the last 50 years, is yet the fourth cause of death for malignant tumor in Italy. Data from the Registro Tumori Umbro di Popolazione have been used to assess incidence, mortality and survival rates for this cancer over the period 1994-1999. The results have been compared with the data of an ad hoc study realized in the period 1978-1982 and with the ones of the other population-based cancer registries. Mortality and incidence rates are among the highest in Italy, confirming the existence of a high risk area for gastric cancer in central Italy; anyway, both mortality and incidence decrease during the study period. Also survival rates from gastric cancer recorded in Umbria over the period 94-99 were among the highest respect to other Italian cancer Registries. A case-control study carried out in a neighbouring area, aimed to explain the elevated incidence of gastric cancer, found that the consumption of a "traditional" diet--rich in proteins (particularly from sausages), nitritis and nitrous compound, alcohol, coffee, and fibres--was an important risk factor for gastric cancer. Survival over the period 94-99 is significantly improved, if compared to 1979-82 survival rate. The high survival rate recorded in the Umbria region could be related both to the number of cases treated in Umbria and to the quality of the regional oncologic system; in fact, in Umbria we reported high survival rates for several malignancies. PMID- 15552733 TI - [Research on Pisa University student's life style and health]. AB - Surveys on life and health conditions of university students besides providing useful epidemiologic data partly related to the young population. They are potentially useful for the intervention planning aimed to remove possible environmental or behavioural risk factors. Following these purpose a survey sample about 1200 student attending the University of Pisa was carried out through an anonymous questionnaire about 41 questions focused on behaviours and lifestyles of young people, possible health problems, self-perception of the health state, the use of public health services, and the more commonly used (or preferred) sources of information on health topics. The results suggest that although the self perception of the state of health is broadly positive for the majority of students, there is also a significant frequency of individuals declaring significant at risk behaviours (persistence of sexual practices at risk, drug use, alcohol and smoke abuse). The consumption, also sporadic, of psicoattive substances has been declared gives beyond 40% of the students, in particular 37% asserts to have tried drugs. The habit to smoke appeared diffused much between the students; the percentage of smokers turns out equal to 41% in total (40% females, 42% males). 76% of the interviewed ones has declared to be sexually active, the percentage of those who have declared multiple relations or occasional is elevated (12%), and 47% of the students asserts to use the condom with fixed partner and 77% with occasional partner. Moreover there seems to be a significant association between the changes in the diet and other lifestyles that are due to the "university lifestyle" (as a consequence of leaving the family or increased commuting) and the insurgence of several types of sickness, e.g. gastroenteric disturbances and fever attacks. Finally, the present investigation suggests that few interventions on services offered to students (such as accommodation, teaching structures and especially the creation of a "health office" where students can acquire information on health and other topics related to their condition) could have a significantly beneficial impact on the general state of health of the university population. PMID- 15552734 TI - Attempted suicides in the municipality of Padua (Italy): a retrospective survey (1996-2000). AB - This study was conduced to determine attempted suicide (AS) rates in Padua between 1996 and 2000, to compare them with Italy and other Veneto areas. MATERIALS AND METHODS: It's an analysis of Police and Carabinieri reports. For each case, they indicate age, sex, marital status, mental and physical disorders, emotional disorders, alcoholism, drug addiction and method of attempted suicide. RESULTS: The annual rate of AS between 1996-2000 ranged from 14.3 (1996) to 3 (2000) per 10(5) inhabitants, with the highest rates in early Autumn; 57.7% of suicide attempts involved females. There is a peak between ages 25 and 34 years, with a higher incidence among females (p < 0.05). Divorce and single status appear to be important risk conditions, especially in females. Mental disorders (especially depression) are the most important condition prior to AS. Both genders have attempted suicide especially by poisoning or falling. CONCLUSION: In Padua AS rates are rather high by comparison with the regional average, but lower than in Verona. Marital status and loneliness (with various age-related peculiarities) influence AS rates and relational problems, inability to overcome frustration (in 10-44 year-olds) may play an important part. The use of educational tools, beginning in the early school years is very important, paying special attention to personal and family distress. PMID- 15552736 TI - Was the NHS set up to be safe? PMID- 15552735 TI - [Cluster of Pseudomonas aeruginosa in a neurosurgical unit]. AB - Hospital infections caused by Pseudomonas aeruginosa and particulary meningitis appear to be very common in neurosurgical wards and in patients with brain traumas. This study was carried out during the period between 25th October and the 11th December 2000 in a neurosurgical ward of the teaching hospital of the University "Federico II" of Naples. During this period, five patients contracted an infection caused by P. aeruginosa. The microorganisim was found to be responsible for three cases of meningitis and two cases of surgical site infections. The P. aeruginosa isolates responsible for the infections and the "environmental" isolates were subjected to genotypic typing through the analysis of macrorestriction patterns of genomic DNA after XbaI digestion of Pulsed Field Gel Electrophoresis (PFGE). Four clinical isolates and an environmental isolate recovered from an hand washing basin located in the sub-intensive care area showed an identical PFGE pattern, as well as the some multiresistant antibiotype. The results of this study allows us to point out that the surveillance programs of infections in hospitals should include the molecular typing of micro-organisms singled out in clinical samples and, in case of outbreaks, also the typing of microorganisms originating from the environment and the hospital staff. PMID- 15552737 TI - Leading by example. PMID- 15552738 TI - Safety culture? What safety culture? PMID- 15552739 TI - Almost the real thing. AB - Simulation training offers tuition that is constructive, realistic and highly participatory, and by representing believable working environments, simulators are ideal for undertaking technical and behavioural training, for individuals, small groups or entire teams. With current government led changes in healthcare provision such as implementing the European working time directive (DoH 2003), healthcare training opportunities with real patients are likely to become scarcer. The challenge will then be to train skilled, knowledgeable and proficient individuals in other ways. Simulation centres should offer part of the solution. PMID- 15552740 TI - On target in A&E. PMID- 15552742 TI - How to lobby. PMID- 15552741 TI - Scaling the political ladder. PMID- 15552743 TI - Integrating complementary and alternative medicine. PMID- 15552744 TI - [Osteoarthritis: a new priority for rheumatology?]. PMID- 15552745 TI - [New treatments for ankylosing spondylitis: re-evaluating certain myths and dogmas]. AB - We propose a revision centered on the primary care physician practice of a few beliefs on frequency, diagnostic, natural history and classical treatment of ankylosing spondylitis as a help to optimize management at a time of availability of new treatments. PMID- 15552746 TI - [Foot orthotics: indication and realization]. AB - Feet problems are a very often reason for searching medical advice: one out every people after 65 years of age have foot problems. Conservative medical treatment associated with insoles, braces and special shoes is a very important part of treatment. Patient with foot problem could have just a simple conflict due to too narrow shoes or be as severe as an destructive diabetic arthropathy. After reminding the great groups of pathologies, causes and effects, we describe successively effects, kinds indication and fabrication of foot orthosis and shoes. Prevention of complications is a very important tool in treating feet problems. Every patient with a foot orthosis should have a very good footwear. The interdisciplinary approach with orthesist is mandatory, as is the precise and meticulous follow-up of this patient. PMID- 15552747 TI - [Efficacy of tumor necrosis factor-alpha blockade for severe sciatica?]. AB - The domain of sciatica is at the edge of a mini revolution. For ten years evidence have been accumulating in favour of a local inflammation rather than a pathology resulling only from a nerve compression. This hypothesis has first been strengthened by the discovery of inflammatory mediators in human herniated discs and then by animal models. These models have demonstrated the impossibility for nerve root compression to produce sciatica in the absence of inflammation and the importance of proinflammatory cytokines in this pathology. TNF-alpha have been proved to be the most important inflammatory cytokine and TNF-alpha modulators has been most effective in the treatment of these models. Two pilot studies realized on humans seem to confirm these experimental data. A multicenter randomised, double-blind, placebo controlled study is being planed in Switzerland. PMID- 15552748 TI - [Review of the protective effects of tumor necrosis factor inhibitors in rheumatoid arthritis]. AB - Rheumatoid arthritis (RA) is a chronic in-flammatory disease with progressive joint damage, generally considered irreversible. A more aggressive therapeutic approach is promoted. Progression of joint damage, gradual functional disability and rate of mortality are lowered by disease modifying antirheumatic drugs (DMARD). Among these, we discuss the relative role of TNF inhibitors and their efficacy on preventing radiographic progression of joint damage. We also touch on the notion of a therapeutic window of opportunity early in the course of RA, as early initiation of aggressive treatment seems to provide a sustained benefit on radiographic progression. PMID- 15552749 TI - [Iatrogenic rheumatic diseases]. AB - This article reviews, through illustrative cases, five rheumatic diseases associated with drug treatments: statins and myopathy, quinolones and tendinopathy, viscosupplementation and pseudoseptic arthritis, diuretics and gout, barbiturates and adhesive capsulitis. Due to the rarity of such cases, we lack validated therapeutic recommendations. Announcement to the adverse drug reaction reporting system is the best way to enhance our knowledge and to improve the public health. PMID- 15552750 TI - [Chondrocalcinosis: a disease frequently occurring in the second half of life]. AB - Articular chondrocalcinosis (ACC) is a metabolic arthropathy caused by calcium pyrophosphate crystal deposits. It is frequent in the second half of the life (6% of the population aged between 60 and 70 years, 30% of the elderly after 80 years). Both males and females, are involved in the same proportion. Clinical manifestations include pseudogout attacks, mainly of the knee and wrist joints, secondary osteoarthritis and destructive arthropathy (mainly of the knee and hip joints). There is no basis therapy. The pyrophosphate calcium deposits do not disappear during the evolution of the disease. Symptomatic therapy with NSAIDS, analgesic drugs or intra-articular corticosteroids may be very useful. Orthopaedic surgery is necessary in cases with severe destructive arthropathy. PMID- 15552751 TI - [The popliteal fossa: an unrecognized junction]. AB - The popliteal fossa is located at an important place of the osteo-articular system near the knee of course but also in strong relationship with the neighboured vascular, muscular and nervous structures. The symptomatology of the Baker's cyst is well known but its physiopathology is less obvious. It is important to remain aware of some unusual conditions that must prompt a differential diagnosis reaching from functional articular conditions over vascular and nervous problems to major tumoral problems. The patient's history and a rigorous clinical examination must remain the main stone of the medical evaluation; however, most situations require complementary diagnostic procedures among which the ultrasonography and/or the MRI represent nowadays the gold standards. It belongs to the physician's skill to use these tools in an efficient way according to the clinical condition. Actually, the Baker's cyst is in most cases secondary to a mechanical or inflammatory intraarticular knee problem; the treatment of this condition will cure the cyst that usually doesn't require any own treatment. PMID- 15552752 TI - [Slow-acting anti-rheumatic agents: recent developments]. AB - Until recently, treatment of osteoarthritis targeted the pain and the disability of the involved joints. It consisted in a mix of analgesics, anti-inflammatory drugs, steroid infiltrations and physical therapies. Usefulness of slow but long acting substances is increasing not only by their action on the symptoms but also because they positively influence the evolution of the disease. Some of them seem to induce a structural effect on the cartilage confirmed by a slow down of the joint space narrowing on successive radiographs of the knees. Presently, only basic matricial precursors such as chondroitin sulfate, glucosamine sulfate and hyaluronic acid are available in the clinic. They act by reducing cartilage destruction and enhancing chondrocyte anabolism. These preliminary results should be confirmed by further studies based on magnetic resonance imaging and surrogate markers of the cartilage matrix. Recent developments in the understanding of the physiopathology of osteoarthritis, including the key role of some cytokines, should allow new therapic ways, individually or in combination. PMID- 15552753 TI - [A practical approach to diagnosing and managing post-menopausal osteoporosis]. AB - Postmenopausal osteoporosis is a major challenge in Medicine. From a simple 10 points questionnaire allowing the detection of the main risks of the disease, different treatment options, based on mineral density, are now available. Treatment should be adapted to age and way of life of the patient. PMID- 15552754 TI - [Rheumatoid arthritis of the cervical spine: surgical management]. AB - Cervical spine involvement in patients suffering from rheumatoid arthritis significantly increases with time. This progression results in C1-C2 instability, vertical subluxation, subaxial spine subluxation or a combination of those three types of instability. It can remain asymptomatic or present with pain and/or neurological symptoms. Surgical treatment could be indicated in the presence of C1-C2 instability greater than 6 mm or even grater than 3 mm if there is associated vertical subluxation. Surgery can be associated with significant mortality and morbidity. In the presence of myelopathy surgical results can be particularly unfavourable with a mortality as high as 50%. It seems therefore important to proceed to surgical stabilisation quite early in order to prevent the onset of neurological involvement. Primary fusion extending to the upper thoracic spine should also be considered in selected patients in order to avoid the onset of caudal instability which can present with late development of progressive myelopathy. PMID- 15552755 TI - [Prevention of heterotopic ossification following prosthetic total hip replacement]. AB - Heterotopic ossification, mostly of the hip, is a frequent complication after reconstructive surgery. Most common prophylactic therapies are non-steroidal anti inflammatory medication (AINS) applied post-operatively and external radiotherapy (RT) administered in the pre- or post-operative setting. Both treatments proved to be efficient in several randomised clinical trials. Prevailing trend regarding RT is a treatment prior to surgery. It is appreciated by both physician and the patient because it is simple and comfortable. In this article, we present the literature linked to the subject together with the benefit and the inconvenience of both therapeutic methods. PMID- 15552756 TI - [Biopsy and surgical treatment of soft tissue tumors]. AB - When a soft tissue tumor is initially misdiagnosed as a simple trauma or a mass, treatment may be adversely affected by the delay in diagnosis or an inappropriate invasive procedure that results in extension of a probable malignant tumor. Patients with soft tissue tumors present a diagnostic challenge to the clinician. A systematic approach to the evaluation and biopsy is necessary in order to minimize diagnostic errors and biopsy complications. With a multidisciplinary approach and adequate surgical management, the prognosis with a five-year survival rate can be excellent. PMID- 15552757 TI - [The role of diagnostic imaging in sarcoma]. AB - A radiologist should be included in the multidisciplinary approach which will allow optimal diagnostic evaluation and treatment of sarcoma. The appropriate use of ultrasound, computed tomography and magnetic resonance imaging plays a significant role in the evaluation of these tumors and their recurrences. The potential role of dynamic magnetic resonance imaging has also been evaluated. A review of diagnostic imaging and their clinical utility is discussed. PMID- 15552758 TI - [Clinical quiz: waldenstrom macroglobulinemia]. PMID- 15552759 TI - The price of value. PMID- 15552760 TI - Public health finance: a conceptual framework. AB - In an attempt to stimulate development of public health finance as a field of practice, policy, and scholarship, this article proposes a working definition of the term "public health finance," embeds it in the context of the maturing literature on the public health system and its infrastructure, and proposes a four-part typology that spans both public-sector and private-sector contributions to the financing of prevention and health promotion. A developmental strategy for the field--in applied research, training and education, and performance standards -is outlined as well. PMID- 15552761 TI - Tracking our investments in public health: what have we learned? AB - A voluntary public health expenditures reporting system existed in the United States for over 25 years, tracking and reporting trends in health department expenditures, revenue streams, functions, and programs. Today, no such system exists and no data are available to help us understand how and where public health dollars are being spent and the trends during good and bad economic times. This article discusses the history of and lessons learned from the former public health expenditures reporting system and more recent demonstration projects that experimented with reporting by essential public health services. The article also explores how what we have learned can be used for developing and implementing a system today to meet public health and public policy needs. PMID- 15552762 TI - Examining public health financing in New York State: a methodology for evaluating local and national public health data. AB - Historically, there has been little national, standardized data collection on the financing of public health. In part this is due to the diverse way in which local public health is organized and funded. The lack of consistent financial data to adequately evaluate performance severely limits the ability of policy makers to set standards for improving quality. However, even in the absence of such data and analysis, policies have been established by local, state, and national governments to reduce public health funding, while at the same time, placing additional responsibilities on local public health. In an effort to understand fiscal trends for local health departments in New York State, the New York State Association of County Health Officials conducted a survey in December 2002 that collected data on spending and funding sources from calendar year 2000 to calendar year 2002. The survey instrument presents a model for state and local public health departments to evaluate their own resources/needs and to promote information exchange within and between localities. With limited resources, local public health officials must make difficult choices about their programs. By encouraging localities to collect uniform financial data sets, public health will be better suited to support its case for stable and sufficient funding. PMID- 15552763 TI - Local public health cost study in Georgia. AB - Development of a uniform cost study methodology for local health department services in Georgia began with a desire to negotiate cost-based reimbursement from Medicaid. Managed care and the Health Insurance Portability and Accountability Act requirements for common coding standards added impetus to Georgia's efforts to document the cost of local public health. With a determination that the result was achievable, the Division of Public Health contracted with a medical practice management firm. What followed included a major team effort with active working groups of county, district, and state representatives. A template was developed by the consultants to compile the cost report and automated applications were installed. Statewide training engaged the local public health workforce and led to a successful pilot project. This article describes the interactive process that led ultimately to the ability of every county in the state to produce a valid cost report. The cost of direct services provided for individuals by county health departments can now be calculated with a cost per relative value unit. The cost report also includes the cost of environmental health, dental health, population-based services and all other local public health services. A brief description of the methodology is presented. PMID- 15552764 TI - Combining the benefits of decision science and financial analysis in public health management: a county-specific budgeting and planning model. AB - State public health agencies are charged with providing and overseeing the management of basic public health services on a population-wide basis. These activities have a re-emphasized focus as a result of the events of September 11, 2001, the subsequent anthrax events, and the continuing importance placed on bioterrorism preparedness, West Nile virus, and emerging infectious diseases (eg, monkeypox, SARS). This has added to the tension that exists in budgeting and planning, given the diverse constituencies that are served in each state. State health agencies must be prepared to allocate finite resources in a more formal manner to be able to provide basic public health services on a routine basis, as well as during outbreaks. This article describes the use of an analytical approach to assist financial analysis that is used for budgeting and planning in a state health agency. The combined benefits of decision science and financial analysis are needed to adequately and appropriately plan and budget to meet the diverse needs of the populations within a state. Health and financial indicators are incorporated into a decision model, based on multicriteria decision theory, that has been employed to acquire information about counties and public health programs areas within a county, that reflect the impact of planning and budgeting efforts. This information can be used to allocate resources, to distribute funds for health care services, and to guide public health finance policy formulation and implementation. PMID- 15552765 TI - The Florida Department of Health and the Florida Association of County Health Department Business Administrators: a model of successful collaboration to sustain operational excellence. AB - The Florida Association of County Health Department Business Administrators, Inc., is an organization of public health business professionals working collaboratively with the state's department of health administrative offices in promoting best practices throughout local county health departments. This article discusses a number of joint initiatives, past and present, that are benefiting public health services in Florida. In addition, mentoring and training, contract management, cost allocation, and financial reporting, among others, are "hot button" topics of interest to all states in promoting improved administration of their public health programs. PMID- 15552766 TI - Understanding the financing and functions of metropolitan health departments: a key to improved public health response. AB - Addressing the financing challenges of the governmental public health system requires a more detailed understanding of the specific role and functions of local health departments, particularly for metropolitan health departments. Many current studies and reports mistakenly aggregate all 3,000 local health departments as a single unit of analysis and, as a result, fail to highlight the increasingly important role of metropolitan health departments. These large departments represent less than 10% of local health departments, but at the same time, serve more than 60% of the US population. The largest 25 metropolitan health departments serve jurisdictions averaging 2 million persons; a larger population than 14 of the states. This article presents a description of the capacity and organizational characteristics of metropolitan health departments, the essential role these agencies play in the preparedness and health disparities areas, and the very different public health financing issues that are raised when the policy focus is on the form of local public health practice serving most of the population. PMID- 15552767 TI - Ways to think about money. AB - Healthy People 2010, the nation's objectives for achieving a higher level of health, includes Goal 23-16: Increase the proportion of federal, tribal, state, and local public health agencies that gather accurate data on public health expenditures, categorized by essential public health service. Taking time to understand the complexities of accounting for public health expenditures, as well as the reasons for doing so, is an essential precursor to taking the needed steps to include economic information in public health policy making and public health systems research. PMID- 15552768 TI - There will never be enough money!!! PMID- 15552770 TI - Practices in public health finance: an investigation of jurisdiction funding patterns and performance. AB - A field of study for public health finance has never been adequately developed. Consequently, very little is known about the relationships, types, and amount of finances that fund the public health system in America. This research was undertaken to build on the sparse knowledge of public health finance by examining the value of performance measurement systems to financial analysis. A correlational study was conducted to examine the associations between public health system performance of the 10 essential public health services and funding patterns of 50 local health departments in a large state. The specific objectives were to investigate if different levels and types of revenues, expenditures, and other demographic variables in a jurisdiction are correlated to performance. Pearson correlation analysis did not conclusively show strong associations; however, statistically significant positive associations primarily between higher levels of performance and jurisdiction taxes per capita were found. PMID- 15552769 TI - Getting what you pay for: public health spending and the performance of essential public health services. AB - Governmental spending in public health varies widely across communities, raising questions about how these differences may affect the availability of essential services and infrastructure. This study used data from local public health systems that participated in the National Public Health Performance Standards Program pilot tests between 1999 and 2001 to examine the association between public health spending and the performance of essential public health services. Results indicated that performance varies significantly with both local and federal spending levels, even after controlling for other system and community characteristics. Some public health services appear more sensitive to these expenditures than others, and all services appear more sensitive to local spending than to state or federal spending. These findings can assist public health decision makers in identifying public health financing priorities during periods of change in the resources available to support local public health infrastructure. PMID- 15552771 TI - Applying principles for outcomes-based contracting in a public health program. AB - A national movement is underway for government agencies and their program implementation partners, such as contractors and grantees, to explicitly demonstrate the benefits acquired from the expenditure of public funds. Given such expectations, agencies have adopted initiatives, such as outcomes-based contracting, as quality improvement tools to facilitate performance improvements and to document results. When using outcomes-based contracting methods, payments are linked to accomplishment of mutually agreed upon results. Outcomes are not defined in terms of what is performed, but on the impact of what has been achieved. This case study documents the implementation of some fundamental principles for outcomes-based contracting in a state health department community partnership program. Results are also presented from an interview of contractors that participated in this new contracting process. Interview objectives were to document the impact of outcomes-based contacting on building collaborations and improving accountability. Results revealed perceptions of a highly collaborative relationship between the agency and contractors where contractors viewed outcomes based contracting as improving accountability by focusing on results, establishing and monitoring performance targets, and facilitating contractor flexibility. Respondents also indicated strongly that under this contracting method, they utilized the funding more effectively by linking it with other community investments. PMID- 15552772 TI - Competencies for public health finance: an initial assessment and recommendations. AB - The purpose of the study in this article was to identify The needs of public health managers with regard to public health finance. A survey of public health practitioners regarding competencies was conducted and a review of course offerings in finance among schools of public health was performed. Most public health practitioners surveyed believe that a broad array of management competencies are required to administer the finances of a public health facility or department. Respondents added 35 competencies to those initially given to them for review. Most added competencies that were more specific than the original competencies or could be viewed as subpoints of the original competencies. Many schools offered no courses specifically addressing public health care finance, with a few offering at most only one public health finance course. All schools offered at least one corporate finance course, and the majority offered two or more courses. We conclude with a number of recommendations for education and competency development, suggesting several next steps that can advance the field of public health's understanding of what managers need to master in public health finance to effectively function as public health managers. PMID- 15552773 TI - Economics of prevention: the public health research agenda. AB - Public health policy makers often focus their attention on the economic evaluation methods (eg, cost-benefit and cost-effectiveness analyses) because of their interest in the economic returns from investment in prevention programs. This article presents a case for the broader applicability of economic theories and methods in development of public health prevention research issues. Public financing, delivery, and regulatory policies are often advocated and used to correct the imperfections in the market for preventive health services. A proper understanding of the incentives and constraints faced by individual agents in the market, however, can improve the effectiveness of these policies in achieving the prevention targets. In developing a research agenda, economics can play a more substantial role beyond the traditional economic evaluation research. PMID- 15552774 TI - The coach in you. PMID- 15552775 TI - The impact of federal funding on local bioterrorism preparedness. PMID- 15552776 TI - Tobacco smoking and cancer: a brief review of recent epidemiological evidence. AB - This report summarises the epidemiological evidence on the association between tobacco smoking and cancer, which was reviewed by an international group of scientists convened by IARC. Studies published since the 1986 IARC Monograph on "Tobacco smoking" provide sufficient evidence to establish a causal association between cigarette smoking and cancer of the nasal cavities and paranasal sinuses, nasopharynx, stomach, liver, kidney (renal cell carcinoma) and uterine cervix, and for adenocarcinoma of the oesophagus and myeloid leukaemia. These sites add to the previously established list of cancers causally associated with cigarette smoking, namely cancer of the lung, oral cavity, pharynx, larynx, oesophagus, pancreas, urinary bladder and renal pelvis. Other forms of tobacco smoking, such as cigars, pipes and bidis, also increase risk for cancer, including cancer of the lung and parts of the upper aerodigestive tract. A meta-analysis of over 50 studies on involuntary smoking among never smokers showed a consistent and statistically significant association between exposure to environmental tobacco smoke and lung cancer risk. Smoking is currently responsible for a third of all cancer deaths in many Western countries. It has been estimated that every other smoker will be killed by tobacco. PMID- 15552777 TI - Impact of low-dose CT on lung cancer screening. AB - Despite advances in therapy, the prognosis of lung cancer remains dismal due to the fact that most cases of lung cancer are diagnosed at advanced stages, when the chance of cure is poor. In cases detected at early stages prognosis is better. Unfortunately, early lung cancer usually causes no symptoms and is, consequently, rarely diagnosed. Therefore, screening for early asymptomatic lung cancer with diagnostic procedures appears promising particularly as risk factors for lung cancer are well known (cigarette smoking, occupational asbestos exposure and others) and screening could, therefore, focus on these risk groups. In the past, screening trials using analysis of sputum cytology and to some extent chest radiography have failed to demonstrate a reduction in lung-cancer mortality with screening, probably due to insufficient sensitivity of these tests for early lung cancer. During the last decade the introduction of spiral computed tomography (CT) has provided a technique with a much higher sensitivity for small lung cancers. Feasibility studies using low-radiation-dose CT demonstrated a high proportion of non-small-cell lung cancer at the initial examination (prevalence) with decreasing numbers of detected cancers at follow-up (incidence). The proportion of early-stage tumors was high both at prevalence and incidence examinations. The rate of invasive procedures for benign lesions was low; most indeterminate lesions could be classified with non-invasive diagnostic approaches. The proportion of interval cancers (cancers diagnosed by symptoms between two screening CT scans) was low. As, however, these one-arm feasibility trials are not appropriate to assess a potential mortality reduction through CT screening, prospective randomised multicenter trials were recently initiated in several countries to analyse the effect of CT screening on lung-cancer mortality. PMID- 15552778 TI - Using molecular markers in sputum for the early detection of lung cancer: a review. AB - Recent exciting advances have been made in identifying potential molecular markers in sputum which may soon be ready for validation trials to indicate high risk for lung cancer, especially central airway squamous cell lung cancer. Hopefully, a set of biomarkers will be identified with higher sensitivity, specificity, and enough lead-time compared to traditional cytopathology to justify endobronchial evaluation and local therapy to help reduce lung-cancer mortality in high-risk patients. PMID- 15552779 TI - Fluorescent bronchoscopy: contribution for lung cancer screening? AB - The majority of early lesions, even when centrally located, is missed by conventional white-light (WL) bronchoscopy. Fluorescent bronchoscopy is a promising tool in localising early malignant changes in the central airways, because it is significantly more sensitive than WL bronchoscopy. Originally, drug induced fluorescence has been used, but today autofluorescence bronchoscopy is more common in relation to its simplicity and advantages. Different systems are available which yield comparable results. However, there are no studies comparing all systems. The disadvantage is the low specificity of the method. Fluorescent bronchoscopy will be more effective if carried out in high-risk patients and/or embedded in a program of preprocedural evaluation of sputum and together with a CT examination of the thorax. But a reduction of mortality as a result of such efforts has not been shown until the present time. PMID- 15552780 TI - Lung cancer screening--where we are in 2004 (take home messages). AB - The best prognosis for lung cancer can be expected by diagnosis at an early stage of the disease. Long-term survival may be improved by increasing the number of early-stage diagnoses. At the present time, three different screening tools for lung cancer are available: Low-dose CT scanning, sputum analysis and fluorescence bronchoscopy. Each of these tools has a different screening target. Low-dose CT scanning focusses on small pulmonary nodules, sputum analysis has the potential of detecting lung cancer of the central airways, and fluorescence bronchoscopy can identify pre-malignancy, carcinoma in situ and minimally invasive squamous cell carcinoma. The best way forward appears to be a combination of all techniques. Sputum analysis can be used to define a better-characterised risk population, and subsequently this population can undergo low-dose CT and fluorescence bronchoscopy. PMID- 15552781 TI - Prognosis in N2 NSCLC. AB - The prognosis of bronchogenic carcinoma in stage III N2 is poor. Five-year survival ranges between 0 and 5%. Lymph-node involvement itself still is the main prognostic factor. Complete lymphadenectomy improves long-term survival in contrast to lymph-node sampling. Recent studies have indicated that the number of involved lymph nodes could be another prognostic factor. It has also been proved that complete lymphadenectomy is necessary for correct staging. This also applies to preoperative staging prior to neoadjuvant treatment. For this reason exact knowledge of lymph-node anatomy and drainage is required. To achieve assessment and comparison of mediastinal staging and of lymphadenectomy, the number of pathologically examined lymph nodes should be documented. Other prognostic factors within N2 stages are age and T stage. Molecular markers are subject to major investigation. A definite clinical relevance, however, could so far not be verified for any of them. PMID- 15552782 TI - Surgical exploration of the mediastinum: mediastinoscopy and intraoperative staging. AB - Lung resection remains the therapy of choice offering the greatest potential for cure in non-spread lung cancer. Prognostic importance of lymph-node involvement has been underlined by several studies. So, exploration of the mediastinum is of major importance for defining the therapeutic strategy in a possibly curative setting. Pre-resectional exploration of the mediastinal lymph-nodal status is mandatory to define tumour stage exactly and establish specific therapy. Cervical mediastinoscopy is the primary diagnostic procedure and remains the gold standard in invasive surgical staging. Complementary, parasternal mediastinoscopy, extended mediastinoscopy, and video-assisted thoracoscopy may be performed. These techniques allow accurate assessment of mediastinal lymph-node involvement, resulting in an appropriate judgement as to resectability and possible treatment options. Different techniques are established for intraoperative exploration and staging. In terms of curative surgery of lung cancer we demand accurate staging which is achieved by systematic and complete Lymph-node dissection. So, individually and dependent on primary tumour site, accurate mediastinal staging of Lung cancer should be performed in combination with definitive lung resection. PMID- 15552783 TI - Mediastinal staging--the role of endobronchial and endo-oesophageaL sonographic guided needle aspiration. AB - Conventional transbronchial needle aspiration (TBNA) is a valuable procedure but remains underutilised. Recently, imaging guidance such as CT fluoroscopy has created considerable interest. As CT fluoroscopy is cumbersome and exposes patients and staff to radiation, endosonographic imaging may be a better choice in providing guidance for TBNA or FNA in enlarged mediastinal Lymph nodes. Real time imaging during the procedure is helpful and increases the yield. PMID- 15552784 TI - FDG PET: advantages for staging the mediastinum? AB - Correctly staging lung cancer is extremely important because treatment options and prognosis differ significantly by stage. In this review, the performance characteristics of positron emission tomography using F-18-fluoro-deoxyglucose (FDG PET) for staging the mediastinum are given in comparison with other non invasive imaging modalities on the basis of current literature data. There are three meta-analyses demonstrating that FDG PET is more accurate than CT for detecting mediastinal metastases. First publications suggest that dual-modality PET-CT may provide a further gain in accuracy. Additionally, whole-body FDG PET provides information on M-staging and prognosis. Data comparing FDG PET with other functional imaging modalities or endoscopic ultrasound are limited. The necessity of a histological confirmation of PET results is still under discussion. PMID- 15552785 TI - Value of FDG PET in the management of NSCLC. AB - Metabolic imaging with positron emission tomography (PET) using 18F-fluoro-2 deoxy-glucose (FDG) has been accepted as an important imaging modality in lung cancer. FDG PET may have important impacts on the management of lung-cancer patients, for instance by improvement of locoregional (mediastinal) and extrathoracic staging (unexpected metastases). Interesting findings have now been reported in the response assessment to induction therapy providing results of greater prognostic significance than that obtained by conventional imaging methods. In the field of thoracic irradiation, FDG PET may provide advantages in terms of reduced toxicity, treatment intensification, better local tumour control and increased survival. PMID- 15552786 TI - Histopathologic evaluation of mediastinal lymph nodes in lung cancer. AB - Regional lymph nodes represent the most frequent metastatic site in lung cancer. During histopathologic assessment of lymph-node involvement, in the presence of gross tumour, one or several HEtE-stained sections will suffice to demonstrate the tumour and its possible extranodal extension. In the absence of macroscopically detectable metastatic tumour growth, the entire node should be submitted for microscopic examination and be cut into 3- to 4-mm slices in the longitudinal or transverse plane. If the node is sliced, care should be taken to process different surfaces for microscopic examination. After neoadjuvant therapy, the percentage of therapy-induced necrosis and the still vital tumour tissue in the dissected lymph nodes should be estimated microscopically. EUS guided fine-needle aspiration with subsequent cytologic examination represents a complementary method in the evaluation of mediastinal lymph-node lesions. The proposed way of histopathologic evaluation of mediastinal lymph nodes tries to reach a high diagnostic yield, and to offer a compromise between theoretical demands and practical feasibility. PMID- 15552787 TI - Mediastinal staging (take home messages). AB - Mediastinal lymph-node involvement is still the major prognostic factor in NSCLC. Currently, cervical mediastinoscopy is the gold standard of mediastinal lymph node staging. However, less invasive methods such as transbronchial or transoesophageal FNA are becoming more popular and might replace or adjunct mediastinoscopy under certain circumstances. It has been clearly shown that FDG PET is more accurate than CT for the detection of mediastinal lymph-node metastases. However, a positive finding on the PET scan implies that these lymph nodes have to be examined by invasive methods (e.g. mediastinoscopy). It has been demonstrated that PET might be useful in order to detect non-symptomatic distant metastases. The method of pathological analysis of the lymph nodes is critical for the correct determination of the N stage. PMID- 15552788 TI - Resected non-small-cell lung cancer stage I/II: indication for adjuvant/neoadjuvant therapy? AB - Complete surgical resection remains the current standard of care for operable patients with stage-I or stage-II non-small-cell lung cancer. However, there is a strong rationale that supports the concept of the addition of systemic therapy to surgery either preoperatively or postoperatively even in patients with early stage disease, as distant relapse continues to be the dominant form of relapse after surgical resection of NSCLC. Earlier trials of adjuvant therapy have yielded mixed results and the survival gains demonstrated have been modest at best. However, recently presented data of randomised phase-III trials showed an absolute survival benefit of 12-15% for patients with completely resected stage IB and stage-II NSCLC receiving adjuvant platinum-based chemotherapy compared with observation alone. These trials provide resounding approval for adjuvant chemotherapy being the new standard of care for patients with early-stage non small-cell lung cancer who have undergone complete resection of the tumour. PMID- 15552789 TI - Non-small-cell lung cancer: multimodality approach in stage-III resectable disease. AB - The long-term results of surgery +/- radiotherapy in patients with operable disease of locally advanced non-small-cell lung cancer are discouraging. In the vast majority, disseminated microscopic disease, resulting in the later occurrence of distant metastases, contributes substantially to this poor long term outcome. The further development of multimodality treatment approaches in randomised trials, including effective systemic therapy, is necessary. These approaches and the current status of multimodality treatment strategies of resectable locally advanced non-small-cell lung cancer are discussed. PMID- 15552790 TI - Surgery after multimodality treatment for non-small-cell lung cancer. AB - Neoadjuvant treatment for Locally advanced non-small-cell lung cancer (NSCLC) stage IIIA and IIIB promises higher resection rates because of a reduction of the primary tumour and sterilisation of mediastinal nodes ("downstaging"). In this study we analyse the perioperative course and the long-term survival of patients with trimodality treatment. Between 03/1991 and 12/2002, 392 patients with NSCLC underwent resection after induction treatment. Included were 266 males and 126 females, age 55.8 +/- 9 (28-74), of whom 218 were stage-IIIA patients, 174 were stage-IIIB patients. Induction treatment included 3 courses of chemotherapy with cisplatin/etoposide or cisplatin/paclitaxel, followed by one course of chemotherapy with cisplatin/etoposide as well as hyperfractionated accelerated radiotherapy of the primary tumour and the mediastinal nodes with 45 Gy, followed by surgery. Before induction treatment all patients underwent mediastinoscopy. In patients with N3 disease mediastinoscopy was repeated before surgery. Resections included 133 pneumonectomies (34%), 15 bilobectomies (4%), 55 sleeve lobectomies (14%), 168 lobectomies (42.5%), 6 segmentectomies (1,5%), and 15 explorative thoracotomies (4%). In-hospital mortality rates amounted to 4.6% (18 patients) while postoperative morbidity ran up to 46% (180 patients). Morbidity and mortality rates were significantly higher in patients with Karnofsky status lower than 80% and patients older than 65 years. Bronchopleural fistulas occurred in 16 patients (3.2%). The protection of the bronchial stump or anastomosis with viable tissue, like pericardial fat, proves to be a significant factor for the reduction of septic complications. For NSCLC, the 5- and 7-year survival rates were 36% and 31%, respectively, for stage IIIA, and 26% for stage IIIB. This intensive trimodality treatment proves to be feasible. Treatment-related toxicities are overall moderate and acceptable. Accurate cardiopulmonary evaluation before surgery and reinforcement of bronchial stump or anastomosis can contribute to reducing complications. Induction treatment demonstrated a "downstaging effect", so that a clear trend for organ-sparing resection was observed. Long-term survival rates for selected groups look very promising when compared to historical controls. PMID- 15552791 TI - Stage-III NSClC: multimodality therapy for inoperable tumours. AB - The combination of radiotherapy and chemotherapy (RCT) has improved the treatment results of patients with inoperable stage-III NSCLC in comparison to irradiation alone and is seen as the standard treatment at present. Concomitant RCT was significantly superior to sequential RCT in three of four studies, so that the simultaneous therapy sequence is seen today as the best modality. Nevertheless, the results, with median survival times of 15-18 months and a 5-year survival rate between 10 and 20%, are still not satisfactory so that further possibilities of optimisation are being looked for: --Can the results of concomitant RCT be improved by an induction chemotherapy or subsequent chemotherapy? --Is there an additional benefit of operative measures for tumours which through the application of RCT get operable? --What is the additional benefit of biological response modifiers applied together with RCT? --Is there a place for RCT of elderly patients or patients in poor general condition? The rationale as well as the initial results shall be discussed in this overview. PMID- 15552792 TI - How can we further improve radiotherapy for stage-III non-small-cell lung cancer? AB - Combined modality treatment in advanced NSCLC has produced some gain in treatment outcome. Local control as addressed by radiotherapy is still a significant site of failure. Doses higher than achieved by conventional conformal radiotherapy are shown to result in better control rates. Volume restriction seems to be the most important issue in dose escalation. Integration of PET imaging into target definition, omission of clinically uninvolved lymph-node areas and measures to decrease set-up and movement uncertainties are explored. Introduction of risk estimation based on dose-volume analysis for dose prescription may further optimise individual treatment. PMID- 15552793 TI - Localized small-cell lung cancer: which type of thoracic radiotherapy and which time schedule. AB - The effect of scheduling combined radio-chemotherapy was studied in randomised trials for limited-disease small-cell lung cancer. The available trials with constant overall treatment time of combination therapy were separated into three classes: (1) same radiotherapy scheme, given early vs. late, (2) split-course radiotherapy, given alternately with chemotherapy, vs. continuous-course radiotherapy given concurrently, (3) radiotherapy, given with altered fractionation and the same concurrent chemotherapy. There was a trend towards a better survival in the group in which early radiotherapy was given concurrently with chemotherapy. A shorter treatment radiotherapy cycle, using hyperfractionated accelerated irradiation during concurrent radio-chemotherapy, was more effective than conventional fractionation, whereas alternating radio chemotherapy schedules were as effective as concurrent schedules, leaving overall treatment time of radio chemotherapy constant. The data show that scheduling of radiotherapy and chemotherapy is important in the treatment of a fast proliferating tumour such as SCLC. Prolonged overall treatment with sequential radiotherapy and chemotherapy should be avoided. And there is every indication that an early onset of radiotherapy, concurrently applied to chemotherapy, is superior to a late onset [odds ratio of death within 2 years: odds ratio = 0.77 (0.60-0.99)]. There is a need to increase the effectiveness of radiotherapy over conventionally fractionated radiotherapy to 45 Gy, and this can be accomplished by using hyperfractionated accelerated radiotherapy. PMID- 15552794 TI - Combined-modality treatment of non-small-cell lung cancer stages I-III (take home messages). AB - Combined-modality approaches including surgery, radiotherapy and chemotherapy have led to a clear improvement of treatment results in localised NSCLC. For stages I and II, postoperative adjuvant chemotherapy may lead to an improved progression-free survival. In stage IIIA, preoperative chemotherapy, respectively radio-chemotherapy, increases the incidence of complete resections. Compared to radio-chemotherapy alone the implementation of surgery in stage IIIA(N2)-patients leads to a better progression-free survival. For stage IIIB simultaneous radio chemotherapy with or without additional sequential chemotherapy has become the new standard of treatment for patients in sufficient clinical condition. In the future, the implementation of further individual selection and prognostic parameters including tumour volume, location of the tumour, response to induction treatment, co-morbidities and molecular tumour analysis may contribute to further individualise the treatment approach. PMID- 15552795 TI - Genome-wide screening for prognosis-predicting genes in early-stage non-small cell lung cancer. AB - In early-stage non-small-cell lung cancer (NSCLC), a substantial proportion of patients can be cured by surgery. Development of distant metastases is the most frequent cause of therapeutic failure. The possibility to accurately predict a patient's risk for developing distant metastasis would help to identify patients that are candidates for further intervention such as conventional adjuvant chemotherapy or experimental drugs. Current molecular biology techniques allow the genome-wide screening for differentially expressed genes; and adequate bioinformatics approaches are developed at a rapid pace to improve prognosis prediction. Genes associated with metastasis do not necessarily play a role in disease pathogenesis but rather reflect the activation of specific signal transduction pathways that are associated with enhanced migration and invasion capability. In our own work, we have identified several genes (e.g. thymosin beta 4, elF4A1), including a novel non-coding RNA (MALAT-1) to be expressed at significantly higher levels in stage-I and stage-II NSCLC primary tumours that subsequently metastasised. As a consequence, patients with high-level expression of these genes were shown to have significantly worse survival compared to patients with low-level expression of these genes. These data support the hypothesis that gene-expression patterns in primary tumours determine the tumours' likelihood to metastasise. In the near future, this information will be used for tailored therapy approaches for patients with early-stage NSCLC. PMID- 15552796 TI - Validation of molecular and immunological factors with predictive importance in lung cancer. AB - Histological classification and staging are cornerstones of diagnosis in lung cancer. Treatment options have been enriched in the last few years by the development of a number of new drugs, and therapy is now increasingly being carried out within multimodal concepts and at earlier stages. Still, outcome of the disease is far from satisfactory and progress in clinical and preclinical research is time-consuming. With the whole variety of potent new therapeutic compounds including classical cytostatics and biological factors at hand, many now believe that a clear improvement of treatment results will be derived from a better understanding of the biology of these tumours and a resulting improvement of diagnosis. Biological factors reflecting the underlying tumour biology and aspects of clinically important pathomechanisms may not only better predict outcome of the disease but also of its treatment, serving as surrogate markers for a more appropriate general intensification of therapy and ideally for specific "targeted" interventions. This article describes the different insights in the biology of these tumours in relation with the representing surrogate markers, and opens routes to possible diagnostic and therapeutic consequences. PMID- 15552797 TI - Tumour-stroma interaction: cancer-associated fibroblasts as novel targets in anti cancer therapy? AB - Stroma cells, together with extracellular matrix components, provide the microenvironment that is pivotal for cancer cell growth, invasion and metastatic progression. Characteristic stroma alterations accompany or even precede the malignant conversion of epithelial cells. Crucial in this process are fibroblasts, also termed myofibroblasts or cancer-associated fibroblasts (CAFs) that are located in the vicinity of the neoplastic epithelial cells. They are able to modify the phenotype of the epithelial cells by direct cell-to-cell contacts, through soluble factors or by modification of extracellular matrix components. Seminal functional studies in various cancer types, including breast, colon, prostate and lung cancer, have confirmed the concept that fibroblasts can determine the fate of the epithelial cell, since they are able to promote malignant conversion as well as to revert tumour cells to a normal phenotype. This review focuses on characteristic changes of fibroblasts in cancer and provides the experimental background elucidating functional properties of CAFs in the carcinogenic process. A possible implication in lung carcinogenesis is emphasised. Finally, a laser-capture- and microarray-based approach is presented, which comprehensively characterises carcinoma-associated fibroblasts in their in vivo environment for the identification of potential targets for anti-cancer therapy. PMID- 15552798 TI - Signal transduction pathways as novel therapy targets in lung cancer. AB - Cytotoxic therapy for lung-cancer patients has only moderately improved during the last decades. Simultaneously, efforts of intensive research to increase our understanding of the molecular basis of lung cancer have been undertaken. The cancer cell has been characterised by several genetic changes that lead to altered cellular functions. In addition, multiple factors of the cancer-cell environment further affect the tumour cell via various receptors and subsequent signaling pathways. The increased knowledge of cellular signaling offers the opportunity to develop novel substances that target specific pathway molecules. In the current review, some of the most essential receptors and signaling pathways involved in lung cancer will be described. In conjunction, examples of novel target-specific agents that have already found their way into clinical trials will be discussed. PMID- 15552799 TI - Molecular targeting in radiotherapy of lung cancer. AB - Molecular targeting is a promising option to increase the radiation response of tumours and to decrease normal tissue reactions, i.e. to achieve therapeutic gain. Molecular targeting substances in themselves are not curative while radiation is a highly efficient cytotoxic agent, with local recurrences often occurring from only few surviving clonogenic cells. High-dose radiotherapy therefore offers optimal conditions to evaluate the potential of specific biology driven drugs for oncology. This review summarises the current status of preclinical and clinical research on combined radiation with examples of molecular targeting substances relevant for the treatment of NSCLC (EGFR, COX-2, VEGFR, KGF, TGF-beta, BBI). PMID- 15552800 TI - New targeted treatments in lung cancer--overview of clinical trials. AB - Combination chemotherapy has been established as the cornerstone of systemic treatment for advanced lung cancer in the last ten to fifteen years. However, improvements with new drug combinations in recent years have been rather small and a general outcome plateau has been reached with one-year survival rates of about 40% and two-year survival rates of less than 15%. Survival over three to four years is still a rare event in this disease, and more and more efforts are being made to develop innovative systemic treatment strategies with mechanisms of action different from conventional cytotoxic drugs. These molecular targeted agents have made a strong move forward in the management of this disease since Gefitinib--a small molecule EGF-receptor tyrosine kinase inhibitor--was registered in 2003 by the FDA and a number of further countries for the third line treatment of non-small-cell lung cancer. Since then, every month findings have been reported about new cellular targets on lung-cancer cells and, consequently, new agents aiming at these molecular targets are being developed, preclinically. Some of these agents have already been tested in the clinics within phase-I, phase-II and some even within randomised phase-III trials. In this review we will try to summarise the current knowledge and data on the clinical activity of these new drugs in lung cancer and to give a perspective on the future for these new treatment principles. The most promising strategies have been aiming at the EGF-receptor family, serum-VEGF and the VEGF-receptor family (VEGF-1 and -2, respectively). Results from pivotal registration trials are expected within the next one or two years for a number of these new drugs, and the standards of care for advanced lung cancer may change dramatically, comparable to what we have seen in other solid tumours such as metastasised breast and colon cancer. PMID- 15552801 TI - Molecular oncology--perspectives in lung cancer. AB - Despite novel therapies in lung cancer treatment the 5-year survival rate still remains poor. Furthermore, screening concepts for early diagnosis, based on conventional sputum cytology and chest radiography, have so far not demonstrated an impact on decreasing lung-cancer mortality. More specific molecular markers allow new insights in the process of lung carcinogenesis. Furthermore they raise the hope that they provide new tools for early diagnosis and screening of high risk individuals, determination of prognosis, and identification of innovative treatments. In this review, these perspectives of molecular targets in lung cancer will be discussed and summarised. Angiogenesis-stimulating factors (VEGF, FGF, MMP, etc.), parameters concerning tumour cell proliferation and apoptosis (EGFR, p53, K-ras, rb, bcl-2, etc.) are well known. Several of these genetic factors have already been investigated, but no single parameter has yet gained a sufficient selectivity regarding prognostic significance or therapeutic efficacy. New aspects in the complex tumour-stroma interaction and the interactive, cross talking signal transduction pathways and recently developed functional genomic approaches, such as DNA microarrays and proteomics might lead to further progress in biological staging models and treatment concepts. PMID- 15552802 TI - Chemotherapy in stage-IV NSCLC. AB - Since cisplatin-based chemotherapy was proven to increase survival in advanced and metastatic NSCLC various new combinations have been tested. The third generation regimens which showed almost comparable efficacy among each other in randomised trials often proved a better response rate and time to progression combined with a remarkable reduction of toxic side effects compared to "classic" combinations, whereas most studies only noted a modest increase in survival. Two drug regimens were more effective than monotherapy but at the expense of significantly increased toxicity, while monotherapy compared to BSC improved quality of life and survival. The novel antifolate Pemetrexed proved comparable activity to docetaxel with significantly reduced toxicities. PMID- 15552803 TI - First-line chemotherapy in metastatic small-cell lung cancer (SCLC). AB - The overall treatment results in metastatic small-cell lung cancer have not been changed in the last decades. The prognosis of the disease is still poor with median survival times of less than one year and nearly no chance of cure. This article intends to summarize the current status of treatment in m-SCLC and especially focuses on the aspects of choice of drugs and efforts of treatment intensification either by dose escalation or shortening of treatment intervals. Furthermore the currently available data about the activity of newer drugs, including taxanes and topoisomerase I inhibitors are reported. These cytostatic agents widen the therapeutic options in the treatment of SCLC and will hopefully improve the outcome of the patients in the next years. PMID- 15552804 TI - Interventional endoscopic treatment. AB - Interventional bronchoscopy has evolved as an integral part of lung-cancer treatment but it is not always used to its full potential. The different methods can provide immediate relief of dyspnea and haemoptysis. Bleeding from central airway tumours can be stopped by coagulation preferably with the argon plasma coagulator. In cases of intraluminally growing tumour masses removal of the malignant tissue is accomplished with the Nd-YAG laser, electrocautery, argon plasma coagulation, cryotherapy or photodynamic therapy. Intramural tumour growth is most efficiently treated with high dose-rate endobronchial brachytherapy. Extrinsic compression or airway wall destruction require the placement of an airway stent. All methods can be combined and complement other palliation methods such as radiation or chemotherapy. PMID- 15552805 TI - Palliative percutaneous radiotherapy in non-small-cell lung cancer. AB - Percutaneous radiotherapy is an effective tool for the palliative treatment of patients with non-small-cell lung cancer (NSCLC). About two thirds of patients experience a notably improvement of symptoms after palliative radiotherapy. A whole variety of very different radiation schedules like a single fraction of 10 Gy, 2 fractions of 8.5 Gy, 10 fractions of 3 Gy, 25 fractions of 2 Gy, and others have been used for palliation. The effects of these different schedules have been compared in a total of 11 randomized trials of which 10 reported survival data and form subject of this review. According to these studies, an increase in total irradiation dose does not substantially prolong median survival, but results in a significant better 1-year survival. A comprehensive review of the data reveals that patients with poor performance status (ECOG score > or = 3) do not benefit from higher doses, but patients with good performance status do benefit. Patients with poor performance status, and patients with large distant tumour burden regardless of their performance status, are efficiently treated by a short course of relatively low dose radiotherapy. Schedules like 2 x 8.5 Gy and 4 x 5 Gy are most appropriate in this situation. For patients with good performance status the choice of the optimal radiation schedule is less clear. Schedules with total doses between 30 and 45 Gy in 2.5-3.0 Gy fractions should be preferred in these situations. PMID- 15552806 TI - Treatment of brain metastases from non-small-cell lung cancer (NSCLC): radiotherapy. AB - Brain metastases occur frequently in lung-cancer patients and are associated with a crucial decrease in prognosis and impairment of Life quality. With improved treatment and earlier diagnosis of primary tumour as well as earlier detection of lesions due to improved neuroradiological diagnosis the incidence is apparently increasing. Whole-brain radiation therapy (WBRT) prolongs median survival from 1 to 3-6 months. One-year survival rate after WBRT is approximately 10-20%. Neurological function could be improved with minimal morbidity. However, long term survival is observed in patients with favourable prognostic factors like controlled primary tumour site, no extracranial disease, good performance status and age <60 years. In these patients individually optimised aggressive treatment strategies are clearly justified. Surgical resection or radiosurgery (RS) combined with adjuvant WBRT prolong survival to approximately 8-11 months. Surgical resection is preferred when rapid relief of increased intracranial pressure is required. The incidence of new brain metastases is low in patients with poor prognostic factors. Palliative RS could be used in these patients to rapidly improve neurological deficits. In locally advanced NSCLC radiosurgery may be used to effectively control brain disease without delay in treatment of the primary tumour site. The role of prophylactic ("elective") cranial irradiation in NSCLC patients as well as the role of combined radiochemotherapy for brain metastases has to be addressed in further clinical trials in the future. PMID- 15552807 TI - Treatment of brain metastases from lung cancer: chemotherapy. AB - Brain metastases are a frequent complication in patients suffering from Lung cancer, and a significant cause of morbidity and mortality. Brain metastases are found in about 10% of patients at the time of diagnosis, and approximately 40% of all patients with lung cancer develop brain metastases during the course of their disease. The prognosis of these patients is rather poor. The standard treatment for brain metastases, so far, has been whole-brain radiation therapy and surgery focussing on symptom palliation. The use of chemotherapy for the treatment of brain metastases has been limited because of a presumed lack of effectiveness due to the blood-brain barrier. However, the importance of the blood-brain barrier is probably overrated in the case of macroscopic metastases or relapsed disease as the blood-brain barrier has already been disrupted at this stage resulting from the newly developed blood vessels not provided with the physiological properties of the common blood-brain barrier. Chemotherapeutic agents initially lipid insoluble or liquor-impermeable can also penetrate into the brain and, therefore, trigger action against tumour cells. A number of clinical trials have demonstrated that brain metastases resulting from both small-cell lung cancer and non-small-cell lung cancer are susceptible to systemic chemotherapy. In small cell lung cancer, cerebral response rates up to 50% were observed even in the second-line situation and were comparable to the response rates observed in the primary tumour. In non-small-cell lung cancer, similar results were achieved. Therefore, it seems justified to further evaluate the significance of chemotherapy compared to whole-brain radiation therapy. Whether chemotherapy alone is superior to whole-brain radiation therapy, or whether the combination of both therapeutic modalities should be preferred for the management of brain metastases, has not yet been proven, and further randomised phase-III studies are clearly needed. Based on the current available data, and the promising response rates in patients with lung cancer, chemotherapy should be considered for the management of brain metastases as part of a multimodality (or "interdisciplinary") treatment concept. PMID- 15552808 TI - Therapy of advanced non-small-cell lung cancer (take home message). AB - Palliative therapy of advanced NSCLC aims at symptom relief, improvement of quality of life and prolongation of survival. This goal is usually achieved by one or more of the following treatment modalities: chemotherapy, radiotherapy, special palliative interventions and general supportive care measures. PMID- 15552809 TI - In vitro drug sensitivity and apoptosis induction in newly diagnosed pediatric acute lymphoblastic leukemia: correlation with overall survival. AB - The present study looked for any associations between in vitro drug sensitivity and clinical outcome in pediatric acute lymphoblastic leukemia (ALL) with the standard drugs used for leukemia therapy. A total of 72 samples were analyzed. In vitro sensitivity to drugs was tested by a methylthiazol-tetrazolium assay in 6 serial fold dilutions. Apoptosis was determined by TUNEL assay and apoptotic index was calculated for each sample. Patients sensitive to prednisone, asparginase, vincristine, and 6-mercapto purine had higher overall survival compared to patients whose tumor cells were resistant to these drugs (p < .01). For the other drugs tested, overall survival did not vary from that of the resistant patients. For doxorubicin, asparginase, vincristine, prednisone combination sensitivity, there was a significant worsening of prognosis from the extremely sensitive patients through an intermediate sensitive group to a most resistant group. The present study thus shows that in vitro drug-sensitivity testing provides significant prognostic information in childhood ALL. PMID- 15552810 TI - A case of acute lymphoblastic leukemia presenting as severe hypercalcemia. AB - A 9-year-old girl presented at the Pediatric Emergency Department with an acute onset of gastrointestinal symptoms due to hypercalcemia. Despite the absence of circulating blast, bone marrow biopsy was diagnostic of acute lymphoblastic leukemia. The hypercalcemia was initially treated with intravenous hydration and furosemide, and later on with bisphosphonates. However, the serum calcium levels normalized only after the beginning of specific chemotherapy. Hypercalcemia represents an emergency in children, and acute leukemia must be considered in its differential diagnosis, even when there are no circulating blasts. PMID- 15552811 TI - Five questions for assessing psychological problems in pediatric patients cured of neoplastic disease. AB - Pediatric patients who have been cured of cancer may have particular unexpressed needs and psychopathological consequences. The authors describe their experience with an apparently well-adjusted 20-year-old girl previously cured of rhabdomyosarcoma, who developed severe psychological distress with serious difficulty in controlling her emotions due to defense mechanisms and massive rationalization. The trauma of tumor and its treatment can be associated with psychopathological disorders that may significantly affect the future life of a person cured of cancer. Doctors must learn to notice telltale signs and take the time to investigate the patient's social life and identify any psychopathological disorders. Experience drawn from pediatric oncology might suggest an approach based on a physician-patient relationship that could provide the means for identifying patients' psychological needs and problems. PMID- 15552812 TI - Hemangiopericytoma in an adolescent girl: a case report. AB - Hemangiopericytoma (HPC) is a rare soft tissue tumor The few published reports account for the little information available on its clinical management. Here the authors report the successful treatment of an adolescent girl with rare HPC of the tongue. After incomplete surgical excision of the tumor she was admitted to the Hematology-Oncology Department and was treated with a 3-drug combination regimen (ifosfamide, actinomycin D, vincristine) for 8 weeks. She achieved partial remission in week 9 based on the magnetic resonance imaging (MRI)findings. Conventional radiation therapy was initiated at week 9 and continued until week 16. At week 20, according to the MRI findings, she achieved complete remission and continuation therapy was initiated. The young girl has been alive without evidence of the disease for the last 3 years of follow-up. In conclusion, the current report indicates that in cases of incomplete surgical excision of the tumor, chemotherapy and radiotherapy seem to be effective. PMID- 15552813 TI - Long-term results of high-dose chemotherapy and autologous stem cell rescue for high-risk neuroblastoma patients: a report of the Spanish working party for BMT in children (Getmon). AB - The authors retrospectively analyzed the long-term outcome of 67 patients over 1 year of age at diagnosis with high-risk neuroblastoma (stage 4 or stage 3 with N myc amplification) who were treated with megatherapy and stem cell rescue from 1984 to 1998. Median age at transplant was 4 years (range 1.6-15 years). The source of cells was peripheral stem cells in 29 and bone marrow in 38 patients. In 12 patients, an in vitro purging of bone marrow harvest was performed. Most patients were conditioned with melphalan, BCNU, and VM-26. After transplant 19 patients received complementary treatment with IL-2 (16) or 13-cis-retinoic acid (3). Six patients (8%) died from transplant-related toxicity and 39 from disease progression. Three patients were alive with active disease at the time of analysis. Nineteen patients are alive and disease-free at a median follow-up of 104 months. Five-year event-free survival is 0.30. Survival of patients who received a purged graft was not significantly better than the rest. Post transplant complementary treatment significantly improved overall and event-free survival (p = .01 and p = .04, respectively). PMID- 15552814 TI - Parvovirus B19-associated transient red cell aplasia in children: the role of bone marrow examination in unusual presentations. AB - Human parvovirus B19 (PV B19) infection in children commonly presents as fifth disease. Transient red cell crisis, the other manifestation of PV B19 infection, is usually reported in children with chronic hemolytic anemia, with a worsening of the anemia. However, this condition may pass unrecognized in children without an underlying hemolytic disorder, since the anemia may be of a short duration and self-limiting. The authors report 3 cases of PV B19-induced transient aplastic in different clinical settings--pancytopenia in one child, during induction phase for acute lymphoblastic leukemia in the second, and fever with joint pains in the third. Treatment for PV B19-induced transient aplastic crisis is essentially supportive. There may be a dilemma in patients on immunosuppressive therapy, since initially it is difficult to distinguish between chronic pure red cell aplasia (a condition where intravenous immunoglobulin therapy is beneficial) and transient aplastic crisis, where supportive red cell transfusions suffice. The patient with leukemia also recovered spontaneously despite being on steroids. In all the 3 patients, the pure red cell aplasia recovered spontaneously without administration of intravenous gammaglobulins. PMID- 15552815 TI - The effects of acute infection on hematological parameters. AB - This study was undertaken to investigate the effects of acute infections (e.g., upper respiratory tract infection, acute gastroenteritis, urinary tract infection) on total blood count, the relation of these effects with acute phase reactants, and the level of improvement in the total blood count after the resolution of acute infection. A total of 113 previously healthy children between the ages of 6 months and 12 years were enrolled in the study. The control group consisted of 43 healthy children with proper age and gender distribution. A total of 55.7% of the patients had a decrease of 0.10-2.40 g/dL in Hb values on the 3rd day of acute infections. The comparisons of the 1st, 3rd, and 15th day Htc, RBC, MCV, MCHC, RDW values of the study and control groups revealed no significant differences. 7he 1st day SI, SIBC, and TS values of the study group were low in majority of the patients. Then they gradually increased, finally reaching at their normal levels on the 15th day. There was no significant difference between the sTfR and sTfR/log ferritin values of the study and control groups. PMID- 15552816 TI - Clinicolaboratory findings and treatment of iron-deficiency anemia in childhood. AB - One of the major causes of anemia in childhood worldwide is iron deficiency. Its prevalence depends mainly on age, being higher in infancy and adolescence. Its etiology varies, but poor iron diet is considered the commonest causative factor. Better tactics may be needed, like the targeted screening of children who belong to high-risk groups, to eradicate childhood iron deficiency. The amount of the body iron regulates its absorption from the gut through mechanisms that are still poorly understood. Early identification of iron deficiency is essential for the prevention not only of anemia but also the numerous and long-term consequences caused by the lack of iron. Many tests are available for the diagnosis of the disease. Some of them seem very promising for the early detection of iron deficiency, but further research is needed before they become widely acceptable in clinical practice. Treatment is based on oral iron salts, which do not have any serious side effects. PMID- 15552817 TI - Analysis of 136 febrile neutropenic episodes in children with cancer: evaluation of treatment effectiveness and cost. AB - In this study, 136 febrile neutropenic episodes were overviewed retrospectively. Factors affecting treatment success and cost were analyzed. Twenty percent of the episodes were microbiologically documented and 51 % of the bacterial isolates were gram negatives. The most commonly used empirical therapies in febrile episodes were the combination of two drugs (58.0%), monotherapy (14.8%), and antibiotics plus fluconazole (20.6 %). In lymphoproliferative tumors duration of fever and discharge from the hospital were longer Administration of the hematopoietic growth factors shortened neither the duration of neutropenia nor fever and hospitalization. Treatment costs were higher in lymphoproliferative tumors, in bacteremia, and in episodes where glycopeptides, antifungal drugs, and hematopoietic growth factors were used. In conclusion, duration of neutropenia was a significant independent predictive factor for duration of fever In the lymphoproliferative tumors, duration of fever was longer and cost of treatment was more than in the solid tumors. PMID- 15552818 TI - Second primary myogenic sarcoma in a patient with bilateral retinoblastoma. AB - Retinoblastoma is the primary ocular malignancy affecting children under 6 years of age. The development of second malignant tumors in survivors of hereditary retinoblastoma is a well-known clinical entity and a major cause of morbidity and mortality. Rhabdomyosarcomas as second primary tumors have been only rarely described. The authors report a patient with bilateral retinoblastoma who developed a myogenic sarcoma of the orbit after 5.5 years of diagnosis. The short latency period may be explained by tumor histology with the contribution of radiotherapy and chemotherapy. The prognosis of second tumors is poor despite aggressive treatment. PMID- 15552819 TI - Subcutaneous administration of protein C concentrate. AB - Children born with severe homozygous protein C deficiency do not survive beyond the neonatal period unless they receive protein C replacement. Protein C concentrate (Baxter Biosciences, Vienna) is usually given intravenously via a central venous catheter The authors describe 2 children in whom protein C concentrate was successfully administered by subcutaneous infusion after they had had recurrent central line infections. Alternate daily doses of 250-350 IU/kg resulted in trough protein C levels of > 25 IU/dL. In the follow-up period of 1-2 years neither child has had a thrombotic episode or purpuric skin lesions, and infusions are managed in the home by their parents. PMID- 15552820 TI - Amputee skiers: lofty ambassadors in the rehabilitation of cancer. AB - Adolescence is characterized by inter-related physiologic and emotional development. It is a particularly vulnerable period of life culminating into emergence as an adult. The majority of malignant bone tumors in pediatrics affect the adolescent patient. As a part of the treatment, amputation may be required for cure. This often creates a "grieving process. " Adjustment to this process is described. At the M. D. Anderson Cancer Center to assist in functional and emotional rehabilitation, an annual ski trip for amputees has been held over the past two decades. The accomplishments of this trip are outlined. Motivated by the success of the program, preadolescent amputees and blind and paraplegic cancer patients were also invited to participate. The adolescent amputee paved the way for rehabilitating other physically challenged cancer patients! PMID- 15552821 TI - Disseminated BCG infection resembling langerhans cell histiocytosis in an infant with severe combined immunodeficiency: a case report. AB - We present a very rare congenital immunologic disease, severe combined immunodeficiency syndrome (SCID) in 6-months-old-boy with prolonged mucocutaneous candidiasis, severe anaemia, skin rash similar to the infiltrative eczema of Langerhans cell histiocytosis (LCH) and subcutaneous nodules with histiocytic infiltration. Laboratory findings show profound absence of humoral and cell mediated immunity. Pathology specimens analysis of subcutaneous nodule revealed numerous S-100 protein and Cd1a negative histiocytes, occupied by BCG intracellular growth. Histopathology and immunohistochemistry confirmed the diagnosis of BCG dissemination. BCG vaccination in infants with SCID can lead to life threatening dissemination, resembling to the infiltrative eczema of LCH and may mislead the clinician. PMID- 15552822 TI - Looking for Work? How to use your age as an asset. PMID- 15552823 TI - The mentoring cycle: a six-phase process for success. PMID- 15552824 TI - When is it OK to ration healthcare? We must rationally and fairly make decisions about life-saving resources. PMID- 15552825 TI - Optimizing patient flow. Improve patient throughput and processes while delivering efficient, high-quality care. PMID- 15552826 TI - The CEO's role in healthcare philanthropy. With their commitment, CEOs ensure the success of development programs. PMID- 15552827 TI - Overcoming barriers in designing a recognition program. Compromise is an important tactic in building an employee reward and recognition program. PMID- 15552828 TI - Every child deserves proper healthcare. Mobile health team cares for uninsured children in central Texas. PMID- 15552829 TI - Pay for performance: obstacles/implications. Despite challenges, pay-for performance programs are here to stay. PMID- 15552830 TI - From practice to academia. A successful transition requires new skills and a different perspective. PMID- 15552831 TI - Adding value with better strategy and ATT Governance. Meeting challenges requires '"continuous governance enhancement". PMID- 15552832 TI - Recruiting new members. Basic strategies can help you increase your chapters membership. PMID- 15552833 TI - [Molecular epidemiology and laboratory diagnosis of Clostridium difficile associated diarrhea]. PMID- 15552834 TI - [Norwalk virus and Noro virus]. AB - Norwalk virus and Noro virus are members of the Caliciviridae. These viruses are morphological similarity in each other and shows small round structure. These viruses also are well known as main pathogens of acute infectious gastroenteritis. Clinical features include an incubation period of 24 of 48 hours and illness period of 18 to 72 hours with vomiting and diarrhea in most patients and high secondary attack rates. Oral transmitted infection occurs contaminated water and foods. In our country, outbreak of Noro virus-related gastroenteritis are reported sometimes in hospital and nursing home from winter to early spring seasons. This article are described to the morphlogy, physical characteristics, epidemiology, and clinical manifestation relating to Norwalk virus and Noro virus. PMID- 15552835 TI - [Comparison of a rapid antigen test and cultures for diagnosis of meningitis in children]. AB - Fourteen pediatric patients diagnosed as bacterial meningitis between August 1997 and April 2002 were enrolled in this study. Both rapid antigen detection test, Slidex Meningite 5 Kit (Biomerieux) and culture were performed using cerebrospinal fluids (CSF). H. influenzae was isolated from 11 samples and was the most frequently isolated bacteria, followed by S. pneumoniae from 4 samples and enteric bacteriae from 2 samples. Five out of six samples with positive result by culture were also positive by the rapid antigen test. Gram-negative rod was identified in smear specimens of CSF from all these 5 samples. Significance of the rapid antigen test should be recognized under drug resistance of those bacteriae are increasing. PMID- 15552836 TI - [A case of inactive Escherichia coli isolated from feces initially suspected to be Shigella boydii subgroup C serotype 14 by conventional biochemical tests in tubes]. AB - A small number of colorless colonies grew from DHL agar of feces culture taken as part of a complete physical for a 42 year-old woman who had lived in Singapore for several years. When cultured for first-stage identification using conventional biochemical tests in tubes for Enterobacteriaceae, such as TSI agar slant and SIM medium, the results for lactose reaction (-), saccharose reaction ( ), gas (very weak +) and motility (-) were obtained, and Shigella spp. was suspected. Serological tests (by serotype) for Shigella spp. were then conducted. As a result, clear C14 agglutination was observed. Based on these results, the isolate was strongly suspected to be Shigella boydii serotype 14, but since the woman had no symptoms of abdominal pain, diarrhea or fever, such identification was still questionable. When further identification test was carried out using Walk-Away 96 and VITEK 2, non-Shigella spp. identification results were obtained. In second-stage identification, xylolytic activity, acetate salt utilization and use of carbon sources in CA (citrate-acetate) medium were checked, all results were positive, and the isolate was ultimately identified as Inactive Escherichia coli. While Shigella spp. and E. coli are taxonomically similar, they are quite different from each other in terms of pathogenicity. Accurate and rapid identification of Shigella spp. is therefore important. PMID- 15552837 TI - [Detection of vancomycin-resistant enterococci by a fully automated microbiology system, RAISUS]. AB - A fully automated microbiology system, RAISUS recently developed (Nissui Pharmaceuticals Co., Ltd., Tokyo) was evaluated for identification of enterococci and for detection of vancomycin-resistant enterococci (VRE). When a total of 124 enterococcal isolates were tested, RAISUS correctly identified 122 (98.4%) isolates. Two isolates resulted in species-identifications disagreed with the reference but agreed as belonging to the genus of Enterococcus. When a total of fifty-seven VRE isolates confirmed to be positive for vanA and/or vanB genes were tested against vancomycin, the current RAISUS susceptibility program version 1.76 could detect 41 (71.9%) isolates of VRE as having > or = 32 microg/ml MIC for vancomycin, but one was intermediate (MIC, 8.0 microg/ml) and the remaining 15 vanB-type isolates were incorrectly interpreted as vancomycin-susceptible (MIC, < or = 4.0 microg/ml). The test program based on the algorism to determine bacterial growth in the presence of vancomycin was developed and evaluated. With this test program, all the VRE isolates positive for vanA and/or vanB genes were identified as being vancomycin-resistant or intermediate interpretation. However, eight of 19 clinical isolates of E. casseliflavus and E. gallinarum intrinsically possessing vanC gene were determined as being < or = 4.0 microg/ml MIC for vancomycin. With the influence of program revision, RAISUS became to incubate the test plate longer than with the current program, but 50% of enterococcal isolates including vancomycin-resistant and vancomycin-susceptible isolates were determined within 5 hour-incubations and 90% were within 9 to 10 hour incubations. With these results, we can conclude that the revised test program for enterococcal isolates could rapidly and correctly identify vancomycin resistance, and will be applicable to the routine susceptibility test in clinical laboratories. PMID- 15552838 TI - [Germline mutation of BRCA1 gene in Polish families with strong aggregation of breast and/or ovarian cancer based on coding sequence analysis using the SSCP method]. AB - The aim of the study was to identify possible BRCA1 germline mutations in Polish families with strong aggregation of breast and/or ovarian cancer. Identification was based on analysis of the entire coding region using SSCP--"Phast System" technique. The study group consisted of 34 women with breast or/and ovarian cancer from families with aggregation of breast and/or ovarian cancer referred from the Oncology Center in Szczecin. Breast and ovarian cancer cases were revealed in 16 families, breast cancer in 17 and ovarian cancer in 1 family. Blood samples for genetic analyses were taken from at least one affected woman from each family. As a rule, samples were taken from the youngest woman. Genomic DNA was prepared from peripheral blood leukocytes with the nonenzymatic rapid method described by Lahiri and Nurnberger. The entire coding region of the BRCA1 gene was screened for the presence of germline mutations by use of Single Stranded Conformation Polymorphism (SSCP) followed by direct sequencing of variants. Primers used for PCR amplification of the BRCA1 gene have been described in the Breast Cancer Information Core (BIC) database. Samples were sequenced with the use of fluorescent dideoxy-chain terminators from the ABI Prism Kit (PE Biosystems) and model 373 automated sequencer (PE Biosystems). Four mutations were found in exon 5 (4/9) and three in exon 11 (3/9) of the BRCA1 gene. The two mutations in exon 20--5382insC (2/9) could not be identified using SSCP and required sequencing. Three BRCA1 abnormalities--C61G, 3819de15 and 5382insC, were the most frequent mutations detected in Polish families with strong aggregation of breast and/or ovarian cancers. These three abnormalities were most frequent in hereditary breast-ovarian cancer syndrome (6/9), followed by hereditary breast cancer-site specific syndrome (3/9). The present study and literature data suggest that the most effective strategy for DNA testing for cancer susceptibility genes is to identify the type and frequency of mutations in families with strong familial aggregation of cancers. PMID- 15552839 TI - [Evaluation of toxic doses of fluorine on expression of collagen genes and synthesis of some collagen proteins in rat skin]. AB - The results of studies carried out over the last few decades indicate a significant involvement of fluoride in connective tissue metabolism. Unfortunately, opinions concerning this issue vary and sometimes are conflicting. Collagen constitutes the main component of the extracellular matrix. Biosynthesis of collagen is a complex, multistage process. Each step in collagen biosynthesis may be affected by exogenous factors. The purpose of the present study was to evaluate the effects of toxic doses of fluoride in drinking water on: 1. The level of COL1A1 gene expression in rat skin; 2. Concentrations of acid-soluble, pepsin-soluble and total collagen isolated from rat skin; 3. Ratio of pepsin soluble to acid-soluble collagen concentrations; 4. Proportions of alpha2(I)/alpha1(I) and beta/alpha1(I) collagen chains in extracts ofpepsin soluble collagen. An attempt was made to determine whether fluoride effects are gender-dependent. The experiment was performed in 108 Wistar rats (males and females). Rats were given 60 mg sodium fluoride/dm3 in drinking during 1, 3 or 6 months. Subsequently, animals were sacrificed and blood, femoral bones and fragments of abdominal skin were taken for analysis. Methods used in the study included potentiometry with F- ion-selective electrode, RT-PCR assay, calorimetry, and SDS-PAGE electrophoresis. The results led to the following conclusions: 1. COL1A1 gene expression in the skin of male rats receiving 60 mg NaF/dm3 in drinking water was significantly reduced after 6 months exposure. 2. Fluoride at doses used in the present study basically does not exert an effect on the concentration of each collagen form in rat skin, ratio of pepsin-soluble to acid-soluble collagen concentrations, and relative proportions of some collagen chains in the pepsin-soluble collagen extract. 3. Male rats were more sensitive to fluoride action in the present study as compared with females. 4. Further research on fluoride penetration to the skin in experimental animals is needed in order to elucidate the relatively low toxicity of fluoride in this tissue. PMID- 15552840 TI - [The influence of some trace elements on bioaccumulation in tissues and bioenergetic metabolism of the edible snail Helix aspersa maxima as determined by HPLC of purine derivatives]. AB - The aim of this work was to determine the bioaccumulation of fluoride and some metals (Cu, Zn, Pb) in tissues of snails under strictly controlled conditions expecting with this approach to verify the hypothesis that snails are suitable for the monitoring of environmental hazards. Additionally, the toxicity of fluorides administered orally on the energy balance of the snail's foot was investigated basing on concentrations of nucleosides, nucleotides and their products measured with high-performance liquid chromatography (HPLC). Two parallel snail cultures were started. The effect of dose on tissue levels of fluoride and metals was studied in the first part of the experiment. The second part served to study the effects of fluoride on energy metabolism of foot muscle (Tab. 1). Quantitation of fluoride and metal levels was done in soft tissues (foot, hepatopancreas) and shells of snails. Qualitative and quantitative analysis of purine compounds was performed in slices of foot. Fluoride concentrations in pulverized shells were measured using an ion-selective electrode. Gas chromatography served to determine fluoride concentrations in soft tissues (hepatopancreas and foot). Concentrations of metals were determined spectrophotometrically. Fluoride and metal content was calculated basing on weight of the pulverized sample. Purines were measured in foot muscle slices with high-performance liquid chromatography (HPLC). Concentrations were adjusted for protein content of sample. Concentrations of the following nucleosides, nucleotides and their products were determined: ATP, ADP, AMP, Ado (adenosine), GTP, GDP, GMP, Guo (guanosine), Hyp (hypoxanthine), IMP, Ino (inosine), Xan (xanthine), Urd (uridine), UA (uric acid), NAD+, and NADP (Fig. 1.). Statistical analysis was done with non-parametric test of Kruskal-Wallis, Mann-Whitney U-test and Spearman Rank Correlation Coefficient. CONCLUSIONS: 1. Accumulation in the shell was significantly increased at the lowest concentration of fluoride, but levels remained below those in the foot or hepatopancreas. It can be inferred that due to low sensitivity, accumulation of fluoride in soft tissues is not a suitable indicator for biomonitoring purposes. Shells seem to be more suited for this aim. 2. Due to low sensitivity, accumulation of metals in soft tissues is not a suitable indicator for biomonitoring purposes. 3. Fluoride had a statistically significant effect on the energy metabolism in muscle, especially on the content of AMP and GMP (Fig. 3, 4). The content of adenylate derivatives was increased (Fig. 3, 4) and phosphorylation of ADP to ATP was inhibited (Fig. 2, 5). 4. An increase in TAN and AEC with 1330 mg F-/kg seems to result from inhibition by fluoride of energy-consuming processes (Fig. 5). 5. In cases of high levels of fluoride it seems reasonable to measure the content of AMP, GMP, Guo or the value of AEC which appear to serve as universal indicators of depressed metabolic function (Fig. 3, 4, 5, 6). PMID- 15552841 TI - [Aortic expression of monocyte chemotactic protein-1 (MCP-1) gene in rabbits with experimental atherosclerosis]. AB - The theory of Ross describes atherosclerosis as a process induced by inflammatory reactions involving cytokines, cell adhesion molecules, and chemokines. The latter have been identified as the principal mediator of cell recruitment into the vascular wall when accumulating monocytes become a source of foam cells. The most potent monocyte attractant among known chemokines is the monocyte chemotactic protein-1 (MCP-1). This protein is synthesized in vivo by cells of the vascular wall and its expression is largely controlled by NF-kB nuclear transcription factor. The importance of inflammation for the induction and progression of atherosclerosis suggests that anti-inflammatory drugs could be a useful modality in this condition. The present work was undertaken to: 1) adapt the RT-PCR technique to measurements of MCP-1 gene expression in rabbit aorta, 2) assess MCP-1 gene expression in rabbit aorta during atherosclerosis induced with a cholesterol-rich diet, 3) evaluate the effect of ibuprofen on MCP-1 gene expression in rabbit aorta during atherosclerosis induced with a cholesterol-rich diet. The study was done in 72 rabbits assigned to eight even groups on the basis of body weight and starting cholesterol and triglyceride concentrations in serum. All rabbits were fed a standard chow. In some groups, the diet was supplemented with cholesterol and/or ibuprofen. Two months later rabbits in four groups, i.e. control (K2), control with ibuprofen (IK2), cholesterol-rich (M2) and cholesterol rich with ibuprofen (IM2) were weighed and blood was sampled for measurements of cholesterol and triglyceride concentrations in serum. The liver, heart, kidneys and adrenals were collected at autopsy and weighed. Additionally, a fragment of the ascending aorta was obtained for RT-PCR. The extent of atherosclerosis in aorta was determined using planimetry. Another month later this procedure was repeated for the remaining groups K3, IK3, M3 and IM3. RT-PCR was applied to measure MCP-1 gene expression in relation to constitutive expression of the GAPDH gene. Significantly lower expression was found in rabbits given ibuprofen (groups IK2, IK3, IM3) as compared with groups K2, K3 and M2 (Tab. 1, Fig. 1). Significantly higher concentrations of cholesterol and triglycerides, as well as liver and adrenal mass indices were revealed in rabbits fed a cholesterol-rich diet with or without ibuprofen, in comparison to groups K2, K3, IK2 and IK3. No atherosclerotic lesions were disclosed in control groups. Atheromatous lesions were demonstrated in rabbits fed a cholesterol-rich diet with or without ibuprofen, occupying more than 60% of the intimal surface. The following conclusions were made: 1) RT-PCR corrected for contamination of RNA samples with genomic DNA is a reliable technique for studying MCP-1 gene expression in rabbit aorta, 2) Three months of cholesterol-rich diet is without effect on MCP-1 gene expression in rabbit aorta, 3) Ibuprofen suppresses MCP-1 gene expression in the aorta without affecting the progression of atherosclerosis induced with the cholesterol-rich diet. PMID- 15552842 TI - [Sorption kinetics of fluoride by artificial and natural hydroxyapatite]. AB - The nature and kinetics of fluoride uptake by hydroxyapatite under various conditions remain the object of interest. This problem was now investigated with an experimental model reproducing as closely as possible the conditions in vivo. The aim of this work was: (1) to study the kinetics of fluoride uptake by natural and artificial hydroxyapatite depending on experimental conditions and to determine the reaction rate constants; (2) to describe the equilibrium of fluoride uptake with adsorption isotherms and develop a best fit mathematical model for the process taking place under various experimental conditions; (3) to determine and compare the capacity for fluoride uptake by natural and artificial hydroxyapatite depending on experimental conditions. Attention has focused on the equilibrium and kinetics of the process of fluoride uptake under conditions as similar to those in a living organism as it is possible to reproduce in vitro. Those conditions represented just one of sixteen various experimental setups differing as to process parameters of the experimental system. The equilibria and kinetics of the experimental system were determined basing on measurements of selected parameters. Adsorption isotherms were obtained experimentally and a best fit mathematical model was developed to describe the process. Additionally, maximal capacity for fluoride uptake was calculated, as well as equilibrium constants, adsorption and desorption rate constants for both hydroxyapatite types. The following conclusions were drawn: 1. Fluoride uptake by artificial and natural hydroxyapatite is a biphasic process essentially independent of conditions in the reaction environment adopted in the present work. The first phase is rapid and does not exceed some 15 minutes. The second phase is much slower and takes place over a period of several dozen hours. 2. Fluoride sorption by both hydroxyapatites is essentially a physico-chemical process which can mathematically be best described with Langmuir and Langmuir-Freundlich adsorption isotherms. Under conditions of equilibrium, the adsorbed substance forms a monolayer on the surface of the sorbent. 3. The binding of fluoride by natural hydroxyapatite is stronger than by its artificial counterpart. 4. The stability of fluoroapatite formed during fluoride uptake by natural hydroxyapatite is greater than that of its artificial counterpart. 5. Natural hydroxyapatite has a relatively large capacity for fluoride ions under experimental conditions adopted in the present work. This capacity exceeds that of artificial hydroxyapatite in spite of smaller specific surface of the natural substance. PMID- 15552843 TI - [The influence of hormonal replacement therapy on bone density in postmenopausal women depending on polymorphism of vitamin D receptor (VDR) and estrogen receptor (ER) genes]. AB - Osteoporosis is still an important health problem in modern societies. The densitometric criterion for the diagnosis of this condition established by WHO in 1994 is bone mass density (BMD) lower than 2.5 standard deviation (SD) from the mean value for young healthy individuals of the same sex. Between 60 and 90% of bone density (quantity of bone tissue in the human skeleton) at the time when growth is terminated is genetically determined. For this reason, genes predisposing to osteoporosis and mechanisms of their activity remain the object of investigations. Among them are genes coding for vitamin D receptor (VDR), estrogen receptor (ER), type I collagen, TGF-beta and IL-6. Diminishing bone density past the age of thirty is a physiologic process. Bone loss averages 0.3 0.6% per year. Acceleration of this process to 1.2-6% per year in postmenopausal women has been attributed to constantly decreasing estrogen concentration. Hence, the gold standard in osteoporosis prevention and treatment includes estrogen progestagen therapy enriched with vitamin D analogues, calcium-rich diet and regular physical exercises. Treatment of osteoporosis can be long and expensive. The condition may lead to disability. Osteoporotic fractures and their complications may be fatal. For these reasons, the chief priority in osteoporosis is prevention. Unfortunately, current diagnostic methods (for detection of osteoporosis and monitoring of treatment) remain unsatisfactory. Molecular techniques offer a promising approach to diagnosis and monitoring of therapy. Additionally, the risk of osteoporosis in 1st degree relatives can be assessed and early prevention can be started. The present study addressed the following questions: 1. Are there differences in spine BMD in untreated women with postmenopausal osteoporosis depending on polymorphism of VDR and ER genes? 2. Does efficacy of treatment (increase in spine BMD) in women with postmenopausal osteoporosis depend on polymorphism of VDR and ER genes? 3. What estrogen concentration is necessary to protect bone tissue depending on the polymorphism of VDR and ER genes? The study group included 44 postmenopausal women aged 44-75 years with primary osteoporosis on cyclic HRT (hormonal replacement therapy). Two hormonal preparations were administered: Systen 50 (Jansen Cilag) transdermal system twice per week between day 1 and 21 of the cycle; Provera (Upjohn) 5 mg tablets daily between day 16 and 27 of the cycle. This therapy was supplemented with vitamin D analogue (Alphacalcidolum, Glaxo-Poznan) orally at 0.25 microg per day, calcium-enriched (1200 mg daily) diet and regular physical exercise. Patients were qualified to the study on the basis of a questionnaire. Women with secondary osteoporosis were excluded. TSH, FT3, and FT4 concentrations, as well as fasting glucose were measured. 24 h glycemia was established in women with elevated glucose levels. Polymorphism of the ER gene was studied with Xba I and Pvu II restrictases. Polymorphism of the VDR gene was studied with Bsm I restrictase. Age and BMI were recorded. Spine BMD was determined with DEXA (Dual Energy X-ray Absorptiometry (Lunar instrument) before treatment and after 12 months of HRT. Serum estradiol concentrations were measured before treatment and after 2 months of HTR. The following conclusions were drawn: 1. There is no connection between VDR and ER gene polymorphism and degree of osteoporosis before treatment. 2. XX, PP and Bb markers or X, P, B alleles are associated with a significant decrease in therapeutic efficacy. Nevertheless, satisfactory results were achieved in each woman with primary osteoporosis. 3. Estradiol concentration in serum before and during HRT does not depend on the polymorphism of VDR and ER genes. PMID- 15552844 TI - [Concentrations of selected bioelements and toxic metals and their influence on health status of children and youth residing in Szczecin]. AB - This study was performed in 174 children (59 boys and 115 girls) residing in Szczecin. The objective was to measure the concentrations of heavy metals--lead and cadmium--and some bioelements--magnesium, zinc, copper, calcium, and iron- and to establish relationships between concentrations of elements in serum, erythrocytes, and hair. Mean concentrations of lead in serum and erythrocytes were 0.003 micromol/L (0.07 microg/dL) and 0.96 micromol/L (2.0 microg/dL), respectively (Table 1). Mean concentrations of cadmium in serum and erythrocytes were 0.006 micromol/L (0.07 microg/dL) and 0.02 micromol/L (0.21 microg/dL), respectively (Table 4). In both cases, the maximum allowable concentrations were not exceeded. In hair, the concentrations exceeded the maximum allowable concentration, reaching 0.006 micromol/g dry mass (1.2 microg/g d.m.) for lead and 0.0008 micromol/g d.m. (0.1 microg/g d.m.) for cadmium (Table 3). A positive correlation was found between the concentration of lead in erythrocytes and hair (r = 0.37, p < 0.000001). Concentrations of magnesium in serum and erythrocytes were below the normal range, but were within norm in hair. Mean concentrations of total magnesium in serum, erythrocytes, and hair were 0.69 mmol/L (1.7 mg/dL), 1.6 mmol/L (3.9 mg/dL), and 0.9 micromol/g d.m. (21.5 microg/g d.m.(, respectively (Table 4). 35 children with hypomagnesemia and elevated concentrations of lead and cadmium were placed on magnesium supplementation with Laktomag B6 preparation. This led to significant reduction in the concentrations of lead and cadmium in erythrocytes and hair. Mean concentration of lead diminished from 0.28 micromol/L (5.8 microg/dL) to 0.17 micromol/L (3.6 microg/dL) (p < 0.03) in erythrocytes and from 0.014 micromol/g d.m. (2.9 microg/g d.m.) to 0.007 micromol/g d.m. (1.5 microg/g d.m.) (p < 0.001) (table 5). Mean concentration of cadmium diminished from 0.02 micromol/L (0.23 microg/dL) to 0.015 micromol/L (0.17 microg/dL) in erythrocytes (p < 0.002) and from 0.009 micromol/g d.m. (0.1 microg/g d.m.) to 0.0009 micromol/g d.m. (0.01 microg/g d.m.) in hair (p < 0.02) (Table 6). Magnesium supplementation caused a significant increase in ionized magnesium in serum and and insignificant increase in erythrocytes and hair. No correlations were found in the examined children between hypomagnesemia, elevated levels of lead and cadmium in blood and hair, and health status. PMID- 15552845 TI - [Selected psychoactive drugs--clinical problems and medical-legal aspects]. AB - The multifarious aspects of psychoactive drug use present a significant challenge to the contemporary analyst. During the first stage of the present experiment, the recovery from human serum and urine of some psychoactive drugs with acidic or basic properties was studied. The efficiency of this process was determined using solutions of drug standards added to serum or urine. Classic liquid-liquid extraction, as well as solid phase extraction methods were compared. The efficiency of recovery was checked using high-performance liquid chromatography (HPLC). The results of this study confirm the usefulness of RP-18 sorbent from Merck and the importance in terms of quantitative analysis of the technique selected for isolation of the xenobiotic from the biological material. The second stage of the experiment was aimed at qualitative determination of some narcotics using thin-layer chromatography (TLC). By stepwise comparison and elimination it was possible to develop an optimal system of chromatographic separation using laminar staining. The proposed system and the conditions for separation ofxenobiotics with six selected elution systems and laminar visualization confirm the feasibility of separating 22 psychoactive drugs. The practical use of the system is limited mainly to screening. Conditions for quantitative analysis of diazepam, tramadol, and pethidine in biological material (serum, urine) using high-performance liquid chromatography, as well as morphine in serum using an immunoenzyme assay have been developed. The procedures have been applied to analysis of narcotics and psychoactive drugs administered prior to anesthesia (morphine, diazepam, pethidine) or for suppression of post-operative pain (morphine, tramadol) in 31 patients of an intensive care unit. 10 ml of blood was drawn at fixed times: 30 minutes prior to surgery (S1), at start of surgery (S2), 60 minutes later (S3), 30 minutes after administration of analgesic (S4), and 60 minutes after administration of analgesic (S5). Urine samples were also collected: immediately after surgery (M1) and 90 minutes after administration of analgesic (M2). PMID- 15552846 TI - [Assessment of mycological and clinical factors on the course and results of treatment of mycotic infections in patients with recurrent onychomycosis]. AB - In spite of excellent results of conservative treatment of onychomycosis, there is still a significant number of patients in whom the dermatophyte infection persists in deep layers of the nail plates. Intensive treatment must be undertaken in some of these patients, depending on their clinical state and extent of the persistent dermatophyte infection of the nails. The aim of this study was to determine fungal factors facilitating the persistence of dermatophytosis in nails and to evaluate clinical data of patients in whom conservative treatment was unsuccessful. The study also concentrated on the effectiveness of various kinds of onychomycosis therapy. A total of 704 patients with onychomycosis underwent therapy between 1995 and 2000. 155 patients were previously treated unsuccessfully with terbinafine or itraconazole. Apart from the method of therapy, the main factors influencing the results of therapy in 549 patients visiting the physician for the first time were: age and location and extent of dermatophyte infection. Observation of patients with recurrent mycosis revealed that repeated monotherapy with terbinafine or itraconazole is the least efficient regimen. Significantly better results were obtained with combined itraconazole and pentoxifylline or itraconazole and topical amorolphine. Undoubtedly, surgery is the most efficient method and almost always successful. Surgery remains burdensome and costly, necessitating not only removal of the nail, but also a monthly course of terbinafine and meticulous topical treatment until nail plate regrows. Analysis of factors being potentially capable of influencing the results of treatment revealed that enzymatic activity of dermatophyte is by far the most important one. When enzymatic activity of different dermatophyte species from patients with recurrent onychomycosis was compared with enzymatic activity of control species from patients who were never treated before, significant and characteristic statistical differences were noticed. Statistically significant differences between both groups were detected regarding activities of beta-glucosidase, lipase, arylamidase and N-acetyl-beta glucosaminidase. Higher activities of these enzymes are characteristic for strains isolated from patients who have been unsuccessfully treated before. Undoubtedly, high enzymatic activity is an important factor favoring survival of dermatophytes in the nail. In conclusion, ineffective pharmacological treatment of fungal infection of nails may depend on both clinical features of the patient and the properties of infecting fungus, in particular on its enzymatic activity. PMID- 15552847 TI - [Evolution of indications for cesarean section between 1991 and 2000 in materials from the Pathology Clinic in the Department of Pregnancy and Labor, Pomeranian Medical University in Szczecin]. AB - The present study was designed to analyze retrospectively the course of 2693 pregnancies and deliveries between 1999 and 2000 at the Department of Pathology of Pregnancy and Labor, Pomeranian Academy of Medicine in Szczecin. Attention was focused on the frequency of cesarean sections including preventive and urgent cases, obstetric and paraobstetric pathologies complicating pregnancy, perinatal and maternal mortality. The efficacy of diagnosing fetal distress in pregnancy and labor by continuous fetal monitoring with cardiotocography versus ultrasonography, in particular umbilical artery Doppler velocimetry, was studied. The material consisted of 2693 pregnant women (1476 (54.8%) primiparous and 1217 (45.2%) multiparous) aged 15-46 years, and their 2802 newborns. Gestational age varied between 23 and 43 weeks. Two groups were formed: I--1268 women delivering by cesarean section between 1991 and 1995; II--1425 women delivering by cesarean section between 1996 and 2000. In 1995/1996, complete ultrasonography supplemented with umbilical artery Doppler flow and intrauterine invasive intervention (cordocentesis, amnioinfusion, amnioreduction, intrauterine fetal blood transfusion) were introduced, as well as screening for diabetes in pregnancy and immunological tests for anticardiolipin antibodies. At that time, the Program of Improved Prenatal Care in Poland was fully implemented. Moreover, the Intensive Therapy of Newborns Uni was established at the Department of Pathology of Pregnancy and Labor. Depending on the moment the decision to perform cesarean section was taken, the prospective group was divided into two groups: antenatal (group P--1036 gravida) and intranatal (group S--1657 parturients). Depending on indications for cesarean section, urgent (N) were distinguished from preventive (Pr) cases. CTG interpretation was performe using qualitative and semi quantitative Fischer's method. CTG recordings during labor were interpreted according to the qualitative method proposed by FIGO in 1987. An analysis of PI, S/D, RI, and flow curve (absence (AEDF) or reversion (REDF) of end-diastolic umbilical artery blood flow) in umbilical artery Doppler velocimetr was performed. A decrease of more than two standard deviations from average valu for several pregnancy time intervals was regarded as pathological. The absence (AEDF) or reversion (REDF) of end-diastolic umbilical artery blood flow was considered to be a high risk factor. The status of newborns was assessed using 5 min Apgar score and parameters o acid-base equilibrium in umbilical artery blood. The following conclusions were drawn: 1. Analysis of cases during the last decade of the 20th century revealed: --increased rate of cesarean sections with a shift in indications from urgent to preventive; --increase in obstetric risk associated with higher percentage of complication during pregnancy; --higher percentage of cesarean sections in premature labor; --correlation between premature labor and increased risk of poor newborn statu --decrease in perinatal mortality of newborns. 2. Analysis of methods for antepartum fetal monitoring revealed: - abnormal CTG patterns and abnormal blood flow in umbilical artery with ab sence or reversion of end-diastolic blood flow the coexistence of which were diagnostically effective for prediction of poor newborn status. 3. Analysis of fetal monitoring methods revealed: --normal intrapartum CTG patterns and normal antepartum blood flow in umbilical artery were markers of good newborn status; - abnormal CTG patterns and abnormal blood flow in umbilical artery had lim ited predictive value regarding poor newborn status. 4. Evolution of indications for cesarean section was multifactorial in character. New and more effective methods for fetal monitoring are needed. PMID- 15552848 TI - [Evaluation of early and late effects of surgical treatment in multivalvular heart diseases]. AB - AIM OF STUDY: Assessment of early and late effects of surgical treatment in multivalvular heart disease. MATERIAL: 186 patients operated at the Department of Cardiac Surgery (Pomeranian Academy of Medicine in Szczecin) between November 1985 and December 1998, 146 (78.5%) of whom were with stage NYHA III and IV. 30 patients (16%) had a history of cardiosurgery. Number of procedures performed: DVR--136, DVR+TVS--26, AVR+MVP--17, AVR+MVP+TVS--7. RESULTS: Early postoperative mortality was 15.6% (29 patients). The risk of early death was associated with the following factors: low cardiac output, cardiac arrest and non-rheumatic etiology of acquired valvular disease. Long-term results were complete in 100% of patients. According to NYHA, statistically significant improvement was noted (average values: 3.02 before and 2.01 after surgery; p < 0.00001). Probability of survival after 5, 7, 10, and 14 years was 77, 69, 61, and 575, respectively. Probability of survival was (1) without hemorrhagic complications 90, 77, 62, 20%, respectively; (2) without thromboembolic episodes 93, 88, 83, 73%, respectively; (3) without infectious endocarditis 98, 97, 97, 97%, respectively; and (4) without perivalvular leak 97, 96, 96, 90%, respectively. Poor long-term prognosis was associated with nonrheumatic etiology of valvular disease, III and IV degrees of NYHA before surgery, persistent atrial fibrillation before operation and low cardiac output during the postoperative period. In the group where repair of damaged valve was done, good long-term results were achieved in 75% of patients with mitral valve and 80% with tricuspid valve pathology. PMID- 15552849 TI - [Significance of anatomic variants of bony surroundings of the internal carotid artery and their significance for lateral surgical approaches to the cavernous sinus]. AB - OBJECTIVE: The central skull base region and the cavernous sinus in particular have traditionally been thought inaccessible to surgical manipulations. Despite significant progress in anatomy, physiology, pathology, imaging and surgical techniques, it is still challenging to explore this part of the skull. The main risk is injury to the internal carotid artery running through the area surrounded by bony structures e.g.: anterior, middle and posterior clinoid processes, optic strut, and petrous bone (Fig. 1, 2). Access to this area requires removal of bony structures but at the same time represents the greatest surgical risk. This microsurgical anatomic study was performed to give surgeons more details on these structures and their relation to the internal carotid artery and venous neighborhood, thus making the operative procedure safer. METHODS: 40 central skull base area specimens were harvested from 20 fresh human cadavers during regular anatomopathologic autopsy. Almost immediately after excision, arterial and venous systems were cannulated and injected with colored acrylic. Microsurgical dissections were performed using a surgical microscope, along with morphometric measurements and photographic documentation of anatomical variants of structures studied. RESULTS: The anterior clinoid process is a structure of complex architecture. In most cases (70%), at least one bony spicula arises from the tip of anterior clinoid and not infrequently passes in a fold of the dura directly under the internal carotid. Bony bridges were observed joining spiculae with similar protrusions of the middle and/or posterior clinoid thought to be especially dangerous during anterior clinoidectomy (Tab. 2, Fig. 2). This observation is in agreement with the literature. 60% of anterior clinoids were pierced by narrow venous canals arising from the anterior cavernous sinus and running through the clinoid space (Fig. 12). They are considered a source of bleeding usually encountered at the very end of anterior clinoid removal. The optic strut seems to be a structure of stable and relatively simple architecture. In 20% of specimens, optic struts were pierced by narrow venous canals arising from the anterior cavernous sinus and running through the clinoid space. The anterior clinoid process and the optic strut happen to be pneumatized. In this case, the pneumatized area is covered with a mucous membrane and opens to the sphenoid sinus or posterior ethmoidal cells (Tab. 2). It was quite surprising that the carotid canal roof was incompletely closed in all samples, although the length of opening was highly variable (Tab. 12, Fig. 1). A relatively thick connective tissue layer covered the opening, usually occupying the area under the Gasserian ganglion i.e. the most medial part of the canal. The greater superficial petrosal nerve was a good landmark for identification of position of the carotid canal in the petrous bone. The canal ran almost directly under the nerve. The hiatus of greater superficial petrosal nerve poorly demarks the area of bone removal lateral to the posterior loop of the internal carotid (Tab. 12). Numerous morphometric measurements of anterior clinoid, optic strut, carotid canal and their anatomical variants give an even greater insight into the area (Tab. 3-13, Fig. 3-11). It is obvious that preoperative CT scanning should be done using thinnest slicing available taking into consideration the scale of measurements presented in this paper. CONCLUSIONS: More details were revealed concerning an area already explored by many researchers. New observations are presented on venous canals passing through the clinoid space. A new insight has been obtained into the area around the roof of the internal carotid canal. The present rich morphometric and photographic documentation should be helpful when dealing with vascular, neoplastic or traumatic lesions of the central skull base. It can also be useful for preoperative planning or training at a neuroanatomy lab. PMID- 15552850 TI - [Assessment of early results of cardiac surgery in patients over 65 years of age]. AB - The aim of the study was determine the influence of age on the outcome of cardiac surgery and reveal risk factors important for the outcome of cardiac surgery in patients over the age of 65 years. Early results of surgical treatment was analyzed in 1741 cases operated in extracorporeal circulation. The patients were divided into two groups. The study group consisted of 375 patients over 65 years of age, while the control group consisted of 1366 patients younger than 65 years. It was shownfound that higher mortality in the study group was associated with low ejection fraction, urgency of surgery, class III and IV of CCS, and BMI > 26. Mortality in the control group was associated with female sex, elevated serum creatinine, bacterial endocarditis, tobacco smoking, urgency of surgery, class III and IV of CCS. Factors significantly influencing mortality in both groups were compared. Mortality in group A in comparison with group B was influenced by low ejection fraction, urgency of surgery, class III and IV of CCS and tobacco smoking. The EuroSCORE was calculated for both groups. It was shown that the EuroSCORE scale allows assessment of operative risk for patients over 65 years of age. Although mortality and morbidity in cardiac surgery are elevated for patients older than 65 years, the outcome is influenced more by risk factors than by age alone. PMID- 15552851 TI - [The effect of reinfusion of mediastinal blood drained after coronary surgery on coagulation, fibrinolysis and volume of blood products transfused]. AB - The aim of the study was to assess the effect of reinfusion of shed mediastinal blood on coagulation and volume of blood products transfused postoperatively. In the first retrospective part of the study, two groups of patients were compared with regard to postoperative blood loss and volume of blood products transfused postoperatively. The study group comprised 260 patients who received drained mediastinal blood. The control group comprised 277 patients who did not receive drained mediastinal blood postoperatively. Volumes of blood lost and transfused were similar in both groups (Tab. 2). The percentage of patients who avoided homologous transfusions was slightly lower in the study group, but the difference did not reach statistical significance (Fig. 1). Two subgroups with drainage higher than usual were compared and no significant differences were found (Tab. 3). In the second prospective part, the effect of postoperative autotransfusion on coagulation and fibrinolysis was studied. The study (autotransfusion) group consisted of 39 patients. The control group consisted of 28 patients. Coagulation tests done in both groups are listed in Tab. 1. Statistically significant differences were disclosed for prothrombin time preoperatively (p = 0.024), activated partial thromboplastin time preoperatively (p = 0.0029), and after 4 (p = 0.0046) and 8 hours (p = 0.0238) from surgery. The most notable differences were found for D-dimers. The level of D-dimers 4 hours after operation and reinfusion of mediastinal blood rose to an average of 22617 ng/mL, in comparison with 3419 ng/mL after 4 hours from surgery in the control group (statistically significant). However, after deduction of D-dimers transfused with mediastinal blood, the mean corrected concentration of D-dimers was 7000 ng/mL and was not significantly higher than in the control group (Tab. 4). The consecutive levels of D-dimers in the study group, though significantly higher than in the control group, tended to decrease, indicating that reinfusion did not contribute to activation of fibrynolysis (Fig. 2). It was therefore concluded that reinfusion of shed mediastinal blood allowed for a slight reduction in the volume of blood products transfused in the study group and did not affect coagulation. PMID- 15552852 TI - [Evaluation of selected factors modifying blood loss after total hip arthroplasty and techniques for the prevention of protracted postoperative bleeding]. AB - The aim of this work was to evaluate blood loss in total hip arthroplasty patients, to determine the effect of some factors on bleeding including age, gender, blood group, body build, bone density, and hip replacement technique, and to develop a method for preventing excessive and protracted postoperative bleeding. Altogether, 59 patients with degenerative hip disease were enrolled. Postoperative bleeding was determined after 12 and 24 hours from surgery by measuring the volume of blood in flasks connected to Redon suction drains. The mean volume of blood lost after hip arthroplasty was 797.3 mL after 12 hours and 1029.4 mL after 24 hours. Total blood loss after surgery was 1257.9 mL. No statistically significant differences in blood loss were noted depending on body weight. In consequence, the percentage of blood lost in relation to total blood volume was determined. This indicator was found to be the only objective measure of blood loss in hip arthroplasty patients. Postoperative bleeding was smaller in patients with higher systolic blood pressure intraoperatively, reduced bone density in the femoral neck and spine, and in females. A tendency to reduced bleeding was observed in the elderly and for shorter surgery times. Blood group and hip replacement technique had no effect on blood loss. PMID- 15552853 TI - [The influence of occlusal interferences on pulp vitality threshold in permanent dentition]. AB - Examination of pulp vitality is a valuable aid in diagnosis and treatment and thus has been in use for a long time. Numerous factors influencing the pain threshold have been found including occlusal interferences. The effect of EOI (Experimental Occlusal Interferences) has been described, while the effect of primary occlusal interference awaits further investigation. The aim of this work was to evaluate the influence of malocclusion (primary occlusal interference) on pulp vitality. At first, the pattern of bite forces in normal occlusion was established. For this purpose, 20 individuals were examined. The following results were found: (a) Bite forces on the right and left side are symmetrical; (b) Total bite forces are distributed as follows: (I) upper half arch: medial incisor 6.3%, lateral incisor 2.9%, canine 3.6%, first premolar 10.4%, second premolar 11.7%, first molar 29.4%, second molar 23.8%; (II) lower half arch: medial incisor 4.1%, lateral incisor 4%, canine 4.8%, first premolar 9%, second premolar 15.1%, first molar 27.3%, second molar 29.7% (Fig. 4). Next, 83 patients with unilateral malocclusion were subjected to clinical examination (orthodontic diagnosis) and measurement of bite forces (with T-Scan system). A computer program was used to establish bite forces (in %) for each tooth. 576 teeth were divided into 9 groups in accordance with the orthodontic diagnosis as follows: class II tendency (n = 126), cross-bite (n = 50), rotated (n = 30), scissors bite (n = 23), class III tendency (n = 19), class II (n = 16), open bite (n = 15), class III (n = 9), normal occlusion (n = 288). Each tooth in malocclusion was assigned a contralateral "control" tooth (Fig. 5). Bite forces acting on teeth in malocclusion were compared with control teeth using Student's t-test (Fig. 6). There is no difference between bite forces of teeth in normal occlusion and malocclusion on the opposite side of dental arches in the same patient, the only exceptions being open and cross-bite. Bite force values obtained in a group with normal occlusion and those reported by Blamphin, Kochanska and Gidzinska Glodkowska formed parallel lines (Fig. 7). Pulp vitality threshold was measured at the buccal and occlusal side in all teeth of the group of 83 patients. Measurements were performed using Ez-Test (Satelec). The results are summarized in Table I. CONCLUSIONS: 1. Bite forces are symmetrical on the right and left side and differ for each tooth. 2. There is no difference between bite forces of teeth in normal occlusion and malocclusion on the opposite side of dental arches in the same patient except for open and cross-bites. 3. Bite forces in patients with unilateral malocclusion are slightly lower than in patients with normal occlusion. 4. There is no difference between pulp vitality in normal occlusion and malocclusion on the opposite side of dental arches in the same patient. 5. Pulp vitality threshold is higher at the buccal than occlusal side. 6. Pulp vitality threshold decreases with age and differs with gender. PMID- 15552854 TI - [Treatment of crossbite with the quadhelix appliance and lower lingual arch to maintain constant lower intermolar width]. AB - The quadhelix appliance is often recommended for the treatment of children with crossbite. It is characterized by high effectiveness, low cost and short treatment time. Its modification with asymmetric arms enables asymmetric expansion of the upper arch. However, no clinical studies confirming such action are known. During maxillary expansion the lower arch tends to follow the maxillary teeth by tipping laterally. This phenomenon hinders crossbite treatment. The aim of the present study was to evaluate the effect of asymmetric arms of the quadhelix appliance during treatment of crossbite and to assess the effectiveness of the lower lingual arch in maintaining lower intermolar width. The study group included 20 children with crossbite. The treatment procedure comprised slow maxillary expansion with the quadhelix appliance. A lower lingual arch activated 1 mm inwards was simultaneously used in order to prevent increase in the lower intermolar width through occlusal contacts. The maxillary intermolar widths were measured between the central fossae of the right and left first permanent upper molars. Mandibular intermolar widths were measured between medial buccal cusps. The mean increase in upper intermolar width was 4.1 mm. The mean change in the lower intermolar width was -0.1 mm and was statistically insignificant. Posterior crossbite was eliminated in 16 children (80%). In 20% of children an improvement (e.g. reduction of the number of teeth in crossbite) was noted. The subtraction sum of the upper and lower intermolar widths was -5.7 mm before treatment and -1.4 mm after treatment. The upper dental arch before treatment had an asymmetric shape in all subjects. The width of the more deficient side was 19.65 mm and the wider side measured 22.3 mm. This difference was statistically highly significant (p < 0.0001). After treatment, the width of the side where the shorter arm was used was 22 mm. The width of the opposite side was 23.7 mm. The difference was statistically significant (p < 0.003). The width of the palate increased on the average by 3.5 mm. The change in overbite was statistically insignificant, enabling safe application of this method in patients with crossbite and open vertical skeletal configuration. The length of the upper arch increased by 0.2 mm (p < 0.01). The average treatment time was 17 weeks, depending on the number of teeth in crossbite before treatment (Rs = 0.58; p < 0.007). Longer treatment times (6-7 months) were reported in other studies describing patients of the same age. The quadhelix with asymmetric lateral arms is recommended for the treatment of crossbite with an asymmetric upper dental arch. The simultaneous application of lower lingual arch prevents lower intermolar expansion, thus shortening the time of treatment. PMID- 15552855 TI - [Evaluation of caries prevention programmes in preschool children]. AB - The aim of the two-year study was to compare the efficacy of two caries prevention approaches in preschool children basing on oral hygiene and dentition. In addition, parents were polled in order to evaluate their attitude towards prophylaxis and treatment of caries and to obtain their opinion on the programme. The first year of the study focused on 4 and 6-year-old children in whom two preventive approaches were applied, i.e. daily toothbrushing under supervision of trained (1) teacher or (2) parents. Since the results of the second approach were unsatisfactory, approach No. 1 was adopted during the second year of study in 3, 4, 5, and 6-year-old children. All children were supplied with a low-fluoride toothpaste and the toothbrush was replaced every three months. Approach No. 1 significantly reduced Pl.I by 89% in 4-year-old children, decreased caries progression in primary and permanent dentition, and reduced the number of filled teeth. A more prominent effect of oral hygiene vs. diet on caries progression in primary teeth was documented. Efficacy of approach No. 2 did not meet expectations. PMID- 15552856 TI - [Evaluation methods of nightguard teeth bleaching with opalescence (ultradent) based on clinical and experimental tests]. AB - In March 1989 Haywood and Heymann presented a new method for bleaching teeth called by them "nightguard vital bleaching". This procedure requires that the patient wears for 6 to 8 hours an individually designed guard filled with a viscous solution of carbamide peroxide. The aim of the present study was to determine the efficiency of the guard bleaching method and to address frequently asked questions concerning local complications and the influence of the preparation on the enamel surface. The scanning electron microscope was used in the experimental part to reveal changes in microhardness and on the surface of the bleached human enamel. The purpose of the clinical part was to determine the efficiency of bleaching, to estimate the frequency of pain and other symptoms and to check the influence of bleaching guard on accumulation of dental plaque and gingival status as reflected by the Plaque Index (Pl.I.) and the Gingival Index (G.I.). It was found that bleaching with 10, 20 or 35% carbamide peroxide reduced the microhardness and modified the morphology of enamel surface proportionally to the concentration of the bleaching agent. It was ascertained that 10% carbamide peroxide effectively bleached teeth and considerably improved the hygiene of the oral cavity and gingival status (Pl.I, G.I.). During bleaching, over half of the patients suffered from local complications--thermal hypersensitivity of teeth and irritation of gums. Some deterioration of the bleaching effect was observed after 12 months in females only. Almost 100% of the patients were satisfied with the results of bleaching and would recommend this procedure to family members. PMID- 15552857 TI - [Efficacy of orthodontic treatment according to the Peer Assessment Rating index]. AB - The work deals with assessment of orthodontic treatment using the Peer Assessment Rating (PAR) index. The objective was to rate the results of orthodontic treatment in the following groups: 1. All patients seen at the Department of Orthodontics, Pomeranian Academy of Medicine; 2. Patients treated by individual dentists at the Department of Orthodontics, Pomeranian Academy of Medicine; 3. Patients with distocclusion; 4. Patients treated with fixed appliances. 1. Rating in patients treated at the Department of Orthodontics, Pomeranian Academy of Medicine Mean reduction in PAR for all patients treated at the Department of Orthodontics was 70%. On this basis, the efficiency of treatment in this group can be termed as high, with 18% of patients classified to "marked improvement", 73% of patients to "improvement" and 9% to "deterioration--no change" groups. This reduction in PAR index is a good result in comparison with published data. 2. Rating in patients treated by individual dentists at the Department of Orthodontics, Pomeranian Academy of Medicine Reduction in PAR ranged from 48% to 83%, depending on the dentist concerned. Orthodontist A achieved the highest reduction (83%), while orthodontists C and E achieved lowest reduction (64% and 48%, respectively). Analysis of patients treated by orthodontists A and C demonstrated that the median PAR for orthodontist A was higher by 5 points than for orthodontist C. 80 malocclusions treated by orthodontist A were difficult to manage. Besides, treatment by orthodontist A was approximately 4 months shorter than by orthodontist C. Patients of orthodontist A had approximately two visits less than patients of orthodontist C. Average costs of treatment for orthodontist A were approximately PLN 500 lower than costs for orthodontist C. In 25% of patients of orthodontist A "marked improvement" was achieved as opposed to only 17% of for orthodontist C. 3. Rating in patients with distocclusion The most significant improvement in this group was achieved with Lehman's appliance (84%). Significant improvement was also noted in patients treated with a two-arch fixed appliance (82%). The lowest reduction in PAR was observed in patients treated with one-arch fixed appliance (64%). High efficiency of treatment with Lehman's appliance or with two-arch fixed appliance in patients with distocclusion is comparable with published results. 4. Rating in patients treated with fixed appliances Patients treated with two-arch fixed appliance achieved higher reduction in PAR than patients treated with one-arch fixed appliance. The main criterion was overcrowding of lower incisors. Results show that treatment with two-arch fixed appliance, in spite of lack of overcrowding of lower incisors, improves the efficiency of treatment. Patients treated with two-arch fixed appliance achieved better qualitative and quantitative results than patients treated with one-arch fixed appliance. In effect, the former patients were managed more effectively. The following conclusions were drawn: 1. The efficiency of treatment at the Department of Orthodontics, Pomeranian Medical University, was high; 2. The efficiency of treatment by orthodontists at the Department of Orthodontics, Pomeranian Medical University, was high; 3. Treatment efficacy for patients with distocclusion was highest with Lehman's appliance and with two-arch fixed appliance; 4. Although costs of treatment were reduced with one-arch fixed appliance, efficiency was lower than for two-arch fixed appliance. Moreover, it is necessary to: 5. Treat abnormalities of occlusion affecting the esthetics of bite, but also abnormalities with importance for normal occlusion that the patient is not aware of, like abnormalities in buccal segments, compression of lower incisors, marked overbite and centerline shift; 6. Control right and left buccal occlusion to the same extent; 7. Treat both jaws using removable and fixed appliances. PMID- 15552858 TI - [Histomorphologic evaluation of upper lip frenum in relation to the method of treating diastema]. AB - The aim of this study was to determine collagen density in the hypertrophic upper lip frenum and scar forming after frenectomy and to compare the results with collagen density in a normal population (autopsy specimens). Differences in collagen density may cause problems in diastema closure and may eventually result in relapse of this condition. The study was done in 47 patients treated for diastema. Histomorphology was performed in 15 patients of this group. Autopsy material was obtained from 27 persons. Histomorphometric assessment of collagen density was done after staining of collagen fibers according to van Gieson. A computerized image analysis system was used for interactive detection of stained collagen fibers on the digitized microscopic image. The percentage of positive staining expressed as ratio of positive area to whole area of the field studied was determined and recorded as an Excel file. Collagen density was increased in the pathologic frenum and in the scar as compared with the normal frenum. The difference was statistically significant. Diastema treatment with frenectomy, fixed appliance and retainer produced more stable results as compared to treatment with a removable appliance without frenectomy. The results suggest that the similar collagen density observed in hypertrophic upper lip frenum and scar after frenetomy may hinder complete closure of diastema and facilitate relapse after treatment. PMID- 15552859 TI - [Nursing support at a rooming-in maternity ward]. AB - The international congress called "Birth Quality-Life Quality" has been a key scientific event in Poland resulting in great changes in perinatal care. Improvements in the quality of care at maternity wards were not limited to purely medical issues. Attention became increasingly focused on psycho-social problems of women after parturition. Emphasis is now directed to support by the doctors and nurses and efforts to acquaint young mothers with the benefits of natural feeding, as well as to prepare them for taking good care of their child. Social support is seen as an important tool in the improvement of care over the mother and child. The present study has classified social support according to J. House as informational, emotional, instrumental and worthing. Informational support consists of methods to cope with specific problems that require delivery of information and teaching of new skills. Emotional support focuses on demonstrating care, winning trust and showing empathy. Instrumental support is reflected by delivery of goods or services to persons in need. Finally, worthing support aims at promoting self-esteem. The aim of this study was (1) to analyze the determinants of informational, emotional, instrumental and worthing support and to compare the needs for different types of support among primiparas and multiparas at an obstetrics ward with the rooming-in system; (2) to study correlations between availability and demand for social support; (3) to determine the expectations of patients as to sources of informational, emotional, instrumental and worthing support and as to methods and means used during delivery of informational support. The study was done in a group of 200 women giving birth at the Department of Obstetrics and Perinatology, Pomeranian Medical University in Szczecin. A special questionnaire was developed to determine the needs for support. A modified Social Support Score was also used. It was found that the demand for all determinants of informational support was greater among primiparas than multiparas. In the lactation part, significant differences were noted for 25 determinants (Tab. 1). Similarly, significant differences were noted for 13 determinants of neonatal care and nursing (Tab. 2) and for 9 determinants of postpartum self-care (Tab. 3). A large demand for most determinants of emotional support was noted in the study and reference groups. Statistically significant differences (p < 0.05) were revealed as to 3 determinants representing feeling of safety by the woman during hospitalization (Tab. 4). It was found that the demand for all determinants of instrumental support was greater among primiparas than multiparas (Tab. 5). There were no significant differences in worthing support needs (Tab. 6). Primiparas significantly more often expected informational and instrumental (p < 0.001), as well as emotional support (p < 0.05) (Tab. 7) in comparison to multiparas. The need for social support at a maternity ward was found to be independent of the available support (Tab. 8). The most frequently expected source of informational support is the nurse/midwife at the maternity ward, as well as the doctor at the ward and the outpatient unit. Worthing and instrumental support is usually expected of the husband and the family. The usual methods of delivering support include discussions, training, and demonstration. Brochures remain the preferred vehicle for delivering support (Tab. 9). PMID- 15552860 TI - [Bibliography of scientific publications of the Pomeranian Medical University in Szczecin for 2001]. PMID- 15552861 TI - [Triptans in migraine: a comparative review of pharmacology, pharmacokinetics]. PMID- 15552862 TI - [Triptans in migraine: from clinical view]. PMID- 15552863 TI - [Meta-analysis of triptan treatment in migraine]. PMID- 15552864 TI - [Side effects of triptans]. PMID- 15552865 TI - [Pathophysiology and new treatment for hereditary neuropathy]. PMID- 15552866 TI - [Treatment of spinal and bulbar muscular atrophy]. PMID- 15552867 TI - [A case of young adult presenting with cerebral infarction caused by homocystinuria]. AB - Homocystinuria is a congenital metabolic disorder, and has been known as life threatening risk factor of vascular disease including ischemic stroke. We report a case of cerebral infarction due to homocystinuria. The patient was a 21-year old woman exhibiting left hemiparesis and a previous history of ectopia lentis. Magnetic resonance imaging showed multiple fresh infarctions in the right frontal and temporal lobes, basal ganglia, corona radiata, and internal capsule. The right common carotid angiogram demonstrated complete occlusion at the origin of the right internal carotid artery. Further investigation clarified increased level of serum methionine and homocysteine and urinary homocystin due to cystathionine beta-synthase deficiency. Homocystinuria was diagnosed as the cause of cerebral infarction. The patient was treated by low methionine diet and administration of folic acid, cobalamin, and aspirin. It should be recognized that some patients with homocystinuria are missed in the neonatal screening for congenital metabolic disorders. Recent studies indicated that the homocysteinemia is one of risk factors of ischemic stroke in the general population as well as in the patients of homocystinuria. We recommend metabolic screening for homocystinuria, when treating a juvenile patient with ischemic stroke of unknown etiology. PMID- 15552868 TI - [Argyrophilic grain disease clinically mimicking Parkinson's disease with dementia: report of an autopsy case]. AB - Argyrophilic grain disease (AGD) is a neurodegenerative dementia, which is neuropathologically characterized by the spindle-or comma-shaped argyrophilic grains scattered in the neuropil of hippocampal area. Several research reports have disclosed the pathological, biochemical and genetic characteristics of AGD, whereas the clinical aspects have not been fully investigated. Here we report an autopsy case of AGD. She developed tremor at age 63, and then developed dyskinesia, rigidity and gait disturbance. Thereafter, she had cognitive impairment and emotional disturbance at age 71, and died of pneumonia at age 76. She was clinically diagnosed as Parkinson's disease with dementia due to the presence of parkinsonism and dementia. Macroscopically, the brain demonstrated mild atrophy, and the weight was 1,240 g. Many argyrophilic grains were found in the hippocampus and amygdala. Coiled bodies and ballooned neurons were also present, while Alzheimer-type neurofibrillary changes were mild, consistent with stage 2 of Braak's classification. This case was neuropathologically diagnosed as AGD. In contrast, no remarkable pathological changes, including neuronal loss and Lewy bodies, were found in the nigra, locus ceruleus and basal nuclei. On the basis of the above-mentioned clinicopathological findings, parkinsonism with dementia is considered to be one of the clinical manifestations of AGD. PMID- 15552869 TI - [Surgical treatment for dissecting aneurysm of the anterior cerebral artery presenting only with headache: a case report]. AB - The authors reported the first case of a dissecting aneurysm of the anterior cerebral artery (ACA) presenting only with headache successfully treated by trapping. A 23-year-old man was admitted to our department due to headache. CT scan and MRI showed neither hemorrhage nor infarction. Angiography demonstrated an aneurysmal dilatation at the right A1 segment with venous pooling which suggested a dissecting aneurysm. We performed trapping and resection of the aneurysm without neurological deficits. The natural history of intracranial dissecting aneurysm without ischemia and/or hemorrhage is still unclear, and therefore it is controversial about the necessity of surgical intervention. Our case and review of the literature suggest that surgical treatment should be considered even in patients without ischemia and/or hemorrhage, if it is possible in consideration of hemodynamic condition. PMID- 15552870 TI - [An autopsy case with cerebral histoplasmoma: case report]. AB - Histoplasma capsulatum infection is, for the most part, asymptomatic or of little clinical consequence. Disseminated infection due to H. capsulatum is rather uncommon. Clinically apparent infection of central nervous system (CNS) is rare, and involves in 10 to 50% of patients with disseminated histoplasmosis. Although CNS histoplasmosis is frequently fetal or only discovered as an autopsy, some patients can be effectively treated with anti-fungal agents. We describe a 44 year-old civil man who is engineering contractor with headache without evidence of systemic infection. Magnetic resonance imaging showed enhancing masses in the third and forth ventricles, right interpeduncular cistern, and right cerebello pontine angle. After biopsy of what was presumed to be a malignant lymphoma, the patient died of rapidly progressive multiple cerebral infarctions. The autopsy revealed the CNS histoplasmoma disseminating systemically. And we finally diagnosed him as histoplasmoma by gene analysis. It was extremely difficult to make a diagnosis based on his physical and radiological findings because it should be included in the differential diagnosis of a well or ring enhanced lesion. It is very important to ask patients about their birthplace, past illness, occupation, and where they had traveled. In the present case, the patient working for the construction has visited many countries including the African Continent and Central America. Clinicians should maintain a high index of suspicion in patients who are from any area endemic for histoplasmosis. The clinical, radiological and pathological features of this infection were reviewed in this report. PMID- 15552871 TI - [A case of cortical infarction showing unilateral facial and hypoglossal paresis]. AB - We describe a case presenting with facial and hypoglossal nerve paresis due to cortical cerebral infarction. A 54-year-old man visited our hospital complaining of sudden episode of dysarthria and facial paresis. Neurological findings revealed tongue deviation to the left and left facial paresis with forehead wrinkling while neither limb paralysis nor sensory impairment was observed. Head CT performed on day 3 after onset revealed a cortical infarction in the right prefrontal gyrus. Symptoms gradually improved with medical management. This patient had isolated facial and hypoglossal nerve paresis without other neurological symptoms. Patients with mild paresis of cranial nerves should be diagnosed carefully, because their paresis could be supranuclear type. PMID- 15552872 TI - [Spinal epidural abscess with persistent increase in cerebrospinal fluid protein: a case study]. AB - A 67-year-old man under hemodialysis treatment developed neck stiffness, fever and conscious disturbances. The patient was infected with Methicilin-resistant Staphylococcus aureus (MRSA) sepsis caused by an infection on a dialysis shunt. On admission, he was diagnosed with bacterial meningoencephalitis and underwent a series of antibiotic chemotherapies. The treatment brought cell count in the cerebrospinal fluid to a subnormal level but his clinical status did not improve. The patient continued to have high level of cerebrospinal fluid protein (898 mg/dl). Cervical MRI demonstrated two abscesses deep in the neck as well as in the epidural region of the cervical spinal cord, from C2 to C5 vertebral levels. Based on these findings, spinal epidural abscess (SEA) was diagnosed. Intensive antibiotic chemotherapy especially targeted for MRSA could eradicate abscesses and improve clinical status. However, persistent high protein level in the cerebrospinal fluid could suggest SEA. PMID- 15552873 TI - [Meningioma on brain MRA using 3D-TOF sequence]. PMID- 15552874 TI - [Various mediators of acute pancreatitis and the mechanism of their activation]. PMID- 15552875 TI - [Clinical classification and pathological physiology of acute pancreatitis]. AB - Acute pancreatitis is a protean disease capable of wide clinical variation, ranging from mild discomfort to severe organ failure. Moreover, the inflammatory process may remain localized in the pancreas, spread to regional tissues, or even involve remote organ systems. This variability in presentation and clinical course has plagued the study and management of acute pancreatitis since its original clinical description. However, precise comprehension about the pathological physiology is required for better treatment. We described about the clinical classification and pathological physiology in the general in this PMID- 15552876 TI - [Defense mechanism to prevent ectopic activation of pancreatic digestive enzymes under physiological conditions and its breakdown in acute pancreatitis]. AB - Independent of the etiology, acute pancreatitis is associated with significant morbidity and the potential for mortality. In most patients, acute pancreatitis follows an uncomplicated or mild course. Recent studies in hereditary pancreatitis have clearly revealed that trypsin is the key enzyme at the onset of pancreatitis. However, there are several defense mechanisms to prevent ectopic activation of trypsin under physiological conditions. If the defense mechanisms failed or activation of trypsin occurred over defense ability, trypsin would activate other digestive enzymes and self-digestion of the pancreas would occur. PMID- 15552877 TI - [Animal experimental models for acute pancreatitis]. AB - In acute pancreatitis, multiple organ failure in the early phase and infectious complications in the late phase are contributors to mortality. To analyze the mechanism of aggravation of acute pancreatitis is to investigate the mechanism of organ dysfunction and infection. As strategy to elucidate the mechanism, various animal experimental models are utilized. Caerulein-induced pancreatitis and bile salt-induced pancreatitis (duct injection model) are frequently employed for mild edematous pancreatitis and severe necrotizing pancreatitis, respectively. It is important to select an appropriate experimental model that corresponds to the purpose of study. PMID- 15552878 TI - [Metabolic disorders of patients with acute pancreatitis: carbohydrate, lipid and protein metabolic disorders]. AB - Acute pancreatitis has been reported in several studies to increase the catabolism and proteolysis of skeletal muscle in comparison with normal controls. Decreased levels of total plasma proteins and rapid turnover proteins and a marked decrease of the ratio of branched-chain to aromatic amino acid further characterize the hypercatabolic state. Significant decreases in plasma essential amino acids, with marked reductions of almost all amino acids in the liver and increased uptake of endogenous amino acids by the skeletal muscle mass, have been reported clinically and experimentally. Gluconeogenesis increases, and glucose clearance and oxidation diminish, leading to glucose intolerance in 40% to 90% of cases. As a consequence, insulin canbe required in as many as 81% of patients. PMID- 15552879 TI - [Epidemiology of acute pancreatitis--incidence by etiology, relapse rate, cause of death and long-term prognosis]. AB - The epidemiology of acute pancreatitis is reported with reference to its incidence by etiology, relapse rate, cause of death and long-term prognosis. According to the national investigation about its epidemiology in 1999, the speculated number of patients was 19,500 in 1998. In 1,688 patients reported in the study, there were 1,051 males and 565 females (1.9:1), and 54.8% of them were of mild type, 19.9% of moderate type and 25.3% of severe type. The incidence peaked in their fifties in males and seventies in females. The three most common causes of acute pancreatitis were alcohol consumption, gallstone and idiopathic disease. The relapse rate of severe acute pancreatitis was 12.5%, and 70% of them relapsed only once. The mortality of severe type due to pancreatitis-related causes was organ failures in 73 % within 2 weeks after its attack, and infectious diseases due to pancreatic infection in 33.3 % after more than three months from its attack. PMID- 15552880 TI - [Role of increase in permeability and circulatory failure in the development of organ dysfunction in severe acute pancreatitis]. AB - Although many factors such as histamine, bradykinin, TNFalpha, IL-1, FDP, thrombin, and PAF are involved in the increase of the vascular permeability, neutrophil infiltration is a major cause of microcirculatory impairment and organ failure in acute pancreatitis. Vasospasm and microthrombi formation due to hypercoagulability are also other cause of deterioration of the microcirculation of the pancreas leading to pancreatic necrosis. PMID- 15552881 TI - [Severe acute pancreatitis and abnormalities in blood coagulation and fibrinolysis system]. AB - In acute pancreatitis, coagulo-fibrinolytic abnormalities may easily result in disseminated intravascular coagulation (DIC). The prognosis will become very poor, once bleeding necrosis or circulatory failure also occur in important internal organs other than the pancreas, leading to multiple organ failure. Changes in blood coagulation and fibrinolysis system may be greatly involved in the severity of acute pancreatitis. In medical treatment, it is necessary to be cautious of predictors of severity, and to perform the early diagnosis of DIC along with quick and positive intensive management. PMID- 15552882 TI - [Role of immunological disorders in the pathophysiology of severe acute pancreatitis]. AB - Severe acute pancreatitis is a disease associated with high mortality, causes of which are multiple organ failure(MOF) in the early course and the development of bacterial infections in the later course. Inflammatory response leading to organ failure continues to be one of the major problem after acute pancreatitis. This review summarizes recent studies that demonstrate the critical role played by inflammatory mediators in acute pancreatitis. Immunological disorder, immune suppression, also plays a role at high risk of sepsis in the development of severe acute pancreatitis. We have to clarify the mechanism of immunological suppression in the development of bacterial infections in severe acute pancreatitis. It is important that the elucidation of key mediators in MOF and the mechanism of immune suppression in the development of bacterial infection could be improved by the prognosis of severe acute pancreatitis. PMID- 15552883 TI - [Role of free radicals in the development of severe acute pancreatitis]. AB - Acute pancreatitis is an inflammatory disease which leads to acinar cell damage, interstitial edema, and hemorrhage. Some patients develop severe acute pancreatitis and result in multiple organ dysfunction syndrome. Acute pancreatitis is initiated by the activation of pancreatic enzyme in acinar cells. Following activation of trypsinogen into trypsin, local inflammation is initiated and activated inflammatory mediators are produced. Polymorphonuclear neutrophils, macrophages, and lymphocytes release lysosomal enzymes, oxygen free radicals, vasoactive substances, and proinflammatory mediators. In the course of the development of acute pancreatitis oxygen-free radicals and their derivatives play an important role as the molecular trigger in constituting lesions in the pancreas. Damaged acinar cells as well as activated neutrophils and macrophages produce large amount of oxygen radicals in acute pancreatitis. The hydrogen peroxide, superoxide, the hydroxyl radical, and singlet oxygen are key elements capable of cellular injury in acute pancreatitis. These highly reactive species cause various reactions, such as destruction of lipid membranes by peroxidation of fatty acids and destruction of lysosomal membranes. The oxygen radicals generated in the circulation might injure the capillary endothelium, and play an important role in accelerating the progression of acute pancreatitis. The imbalance of oxygen radical generating and oxygen radical scavenging processes is considered to lead to the cell injury in acute pancreatitis. These oxygen radicals are not only restricted in the pancreatic tissue, but involved in the systemic manifestation of the disease, particularly in the lungs, liver, and blood. PMID- 15552884 TI - [Role of apoptosis in severe acute pancreatitis]. AB - The role of apoptosis in a severe acute pancreatitis is recently given the most attention, but it is hardly known in a clinical study. We show three assumptions between apoptosis and the pathophysiological mechanisms of severe acute pancreatitis in this paragraph. First, the apoptosis of pancreatic acinar cell might prevent from a worsening in acute pancreatitis due to diminish leakage of pancreatic enzyme and extension of inflammation. Secondly, impairment cellular immunity due to peripheral lymphocyte apoptosis might be linked to the development of subsequent infectious complications in acute pancreatitis. Third, apoptosis in various major organs might be involved in the mechanism of multiple organs failure in acute pancreatitis. These findings might lead the new development in the therapeutic strategy of acute pancreatitis. PMID- 15552885 TI - [Diagnostic criteria for acute pancreatitis and evaluation of the severity]. AB - A research group supported by grants from the Ministry of Health and Labor has recently proposed new diagnostic criteria of acute pancreatitis and scoring system to predict the severity of the disease. Comparisons with the present criteria and scoring system were described and their potential problems were briefly discussed. PMID- 15552886 TI - [Recent advances in biochemical diagnosis and assessment of severity in acute pancreatitis]. AB - Serum amylase is most commonly used as a biochemical marker of acute pancreatitis (AP). But it lacks specificity. The serum lipase level is more accurate and a better marker. Serum elastase -1 level is specific and remains elevated longer, but its radioimmunoassay is not routinely used. Recently, it can be rapidly measured by latex turbidometric immunoassay with automatic analyzer. Biochemically, only CRP test is available and useful to assess severity, but its sensitivity is unacceptably low in the early course of the disease. Urinary trypsinogen activation peptide (TAP) or trypsinogen-2 is an earlier marker. Increasing knowledge of the inflammatory process in AP has led to possibly useful biochemical indicators of severity, such as cytokines, nonpancreatic synovial type group II PLA2 or granulocyte elastase. PMID- 15552887 TI - [Imaging of acute pancreatitis--the role of US, CT and MRI]. AB - Imaging of the pancreas with US and CT plays a major role in the diagnosis and evaluations of patients with acute pancreatitis. In severe acute pancreatitis, dynamic CT is essential to assess the severity of the disease and evaluate the complications. MRI is comparable to CT in its capacity to provide precise information about the severity of acute pancreatitis. MRI has some advantages over CT in being free from ionizing radiation and lower toxity of gadolinium which is used for contrast medium intravenously. The disadvantages of MRI, however, include cost, longer exam duration, limited availability and the practical difficulties of scanning a patient with ICU equipment. PMID- 15552888 TI - [Essential therapeutic strategies for acute pancreatitis--guidelines for initial treatment and their significance]. AB - Acute pancreatitis is a disease with a wide variety of pathophysiologies, ranging from mild to severe condition. In about 80% to 90% of cases, acute pancreatitis presents as a mild inflammation with low morbidity and mortality, self-reversing to normal condition within 3-4 days. However, the natural course of severe pancreatitis progresses into SIRS and necrosis of the pancreas and its surrounding tissue. And infection of the necrotic tissue develops in sepsis and organ failures. Therefore, the initial management for acute pancreatitis would significantly contribute on the early prognosis. The first step includes not only diagnosis but also initial treatment according to etiological assessment and severity stratification. The summarization of initial management revealed as follows: monitorings of temperature, pulse rate, blood pressure, urine output volume, abdominal pain etc, and treatments of cardio-pulmonary care with sufficient fluid resuscitation, pain control, resting of pancreas etc. As the treatment of the initial management after initial onset, the significances of nasogastric tube drainage, prophylactic use of broadspectrum antibiotics, continuous infusion of protease inhibitors, use of H2 receptor antagonists, control by enteral nutrition and so on have been discussed. And also the indications of total parenteral nutrition, selective digestive decontamination and the efficacies of peritoneal lavage, continuous hemodiafiltration and continuous arterial infusion of protease inhibitor and antibiotics have been reviewed. For those events, the evidences were collected by a systemic search of MEDLINE and Japan Centra Revues Medicina. And then practical recommendations were also graded and introduced to provide a framework for clinitians to manage acute pancreatitis as a guideline. This paper described a part of those recommendations. PMID- 15552889 TI - [Treatment of acute pancreatitis with protease inhibitor, H2 receptor antagonist and somatostatin analogue]. AB - Acute pancreatitis sometimes develops to severe condition with a variety of clinical manifestations and high mortality. Autodigestion of the pancreas, secondary to the activation of digestive enzymes, plays a major role in the pathogenetic mechanism of acute pancreatitis. To improve the mortality and complication rates, appropriate treatments based on the precise prediction of disease severity are required. To this end, the early administration of protease inhibitors has commonly been employed for the therapy of acute pancreatitis in Japan. However, a number of clinical trials have failed to show the clinical effects of protease inhibitors, H2 receptor antagonist and somatostatin analogue on the treatment of acute pancreatitis. To evidence the therapeutic value of these agents for acute pancreatitis, well-organized clinical studies will be required. PMID- 15552890 TI - [Efficacy of selective digestive decontamination (SDD) for severe acute pancreatitis]. AB - Bacterial translocation (BT) is involved in the development of pancreatic infection in severe acute pancreatitis (SAP) and influences the prognosis. We should suppress BT to prevent pancreatic infection and improve survival rate. Selective digestive decontamination (SDD) is rational treatment. We should aim at effective SDD, ensuring the following 4 points: (1) enteral antimicrobials, in combination with, (2) parenteral antibiotics given immediately on admission, (3) hand hygiene, (4) surveillance cultures of throat and rectum. Starting enteral feeding as soon as possible and avoiding long-term SDD is useful for maintaining the defenses of the intestinal mucosa and preventing emergence of resistant bacteria. We used glutamine and dietary fiber besides SDD and were able to reduce pancreatic infection. PMID- 15552891 TI - [New aspects of pharmaco-therapy for acute pancreatitis]. AB - Despite improvement in intensive medical care management, the mortality rate from severe acute pancreatitis is still high. Attempt to reduce the mortality rate, some new drugs were investigated on experimental pancreatitis and clinical cases. There have been several clinical trials of somatostatin, somatostatin analogue octreotide, and a cholecystokinin A(CCK-A) receptor antagonist, loxiglumide, for the inhibition of pancreatic secretion. On the other hand, for the inhibition of the systemic inflammatory response, many studies in the use of the platelet activating factor(PAF) antagonist, lexipafant, were reported in clinical trials. IS-741, a new synthetic anti-inflammatory agent, has been studied on various models of experimental pancreatitis. In this paper, it is introduced the results of these new drugs on experimental pancreatitis or clinical trials. PMID- 15552892 TI - [Nutritional therapy in acute pancreatitis]. AB - In this review, systematic search of literatures for acute pancreatitis and nutrition was performed. Eleven randomized controlled trials (RCT) were found. Eight of them are about parenteral or enteral nutrition, 2 are about immunomodulated nutrition, and one is nutritional method in postoperative period. None of these showed benefit of total parenteral nutrition. Moreover, enteral nutrition via nasojejunal tube after 1-2 days after onset or operation of acute pancreatitis showed safe and more effectiveness than parenteral nutrition even in severe cases. These results indicate no benefit of parenteral nutrition in mild to moderate pancreatitis. Early enteral nutrition via nasojejunal tube can be used even in severe cases without ileus or intestinal ischemia. PMID- 15552893 TI - [Indications for and efficiency of peritoneal lavage in severe acute pancreatitis]. AB - Peritoneal lavage was developed as a simple method to evacuate the ascitic fluid associated with pancreatitis. Although a number of studies demonstrated its efficacy in improvement of clinical manifestations, a meta-analysis of randomized control studies could not reveal its effectiveness on mortality or morbidity in severe acute pancreatitis. However, it has been well established that the ascitic fluid with pancreatitis is extremely toxic and to induce organ damage due to apoptotic cell death. Particularly, we have recently found that the ascitic fluid with pancreatitis can induce bacterial translocation due to the increase of gut permeability. Therapeutic strategy of peritoneal lavage, i.e. evacuation of toxic substances from peritoneal cavity, should be reevaluated to achieve further improvement of treatment result of severe acute pancreatitis. PMID- 15552894 TI - [Indications for and clinical efficacy of continuous hemodiafiltration (CHDF) for severe acute pancreatitis]. AB - It has been claimed humoral mediator network including pro-inflammatory cytokines plays a pivotal role in the pathophysiology of SAP. On the other hands, we have reported CHDF using polymethyl methacrylate (PMMA) membrane hemofilter could remove cytokines in blood of a patient continuously and effectively. Therefore, we applied CHDF to patients with SAP. CHDF was performed using a PMMA-hemofilter for removal of causative cytokines regardless of renal function 24 hours a day without interruption until IL-6 blood level became below 400 pg/ml. Selective digestive decontamination (SDD) was also given for prevention of bacterial translocation. Forty-two out of 45 patients (93%) receiving both CHDF and SDD survived. We conclude critical care with PMMA-CHDF and SDD is an effective treatment for SAP. PMID- 15552895 TI - [Continuous regional arterial infusion of protease inhibitor and antibiotic for severe acute pancreatitis]. AB - Continuous regional arterial infusion (CRAI) of protease inhibitor and antibiotic has been established as an initial treatment for severe acute pancreatitis in Japan. CRAI therapy reduced the mortality rate and the frequency of infected pancreatic necrosis. Recently vasospasm has been recognized to be involved in the development of pancreatic ischemia or hypoperfusion in the early phase of the disease, and early application of CRAI of nafamostat may protect the evolution of ischemia into necrosis by inhibition of the increase in the intravascular coagulant activity. PMID- 15552896 TI - [Surgical indications and timing in severe acute pancreatitis]. AB - Infected pancreatic necrosis is the major risk factor with regard to morbidity and mortality in severe acute pancreatitis. Whereas surgery for sterile necrosis can only be recommended in selected cases, infected pancreatic necrosis is a well accepted indication for surgery. Surgery should be postponed until 4 weeks after the onset of symptoms, as necrosis is well demarcated at that time. Two surgical techniques can be performed with comparable results regarding mortality: necrosectomy combined with closed continuous local lavage or open drainage. Selection of these techniques depends on the extent of infected peripancreatic fat necrosis. Open drainage seems to be selected for the cases in which infected tissue is extensively found. However, local lavage is associated with the lower morbidity than open drainage. PMID- 15552897 TI - [Anti-cytokine therapy for severe acute pancreatitis]. AB - The systemic manifestations of acute pancreatitis (AP) are responsible for the majority of pancreatitis-associated morbidity and mortality. Recent studies have established that severe AP is a disease with systemic inflammatory response syndrome as well as compensatory anti-inflammatory response syndrome. Based on their roles in the pathogenesis of AP, new therapies have been sought and tested, including those preventing the biological activity of two pro-inflammatory cytokines, tumor necrosis factor (TNF)-alpha and interleukin-1 (IL-1). Biological activity of TNF might be blunted by anti-TNF-alpha antibody or soluble TNF receptor, and IL-1 receptor antagonist might blunt that of IL-1. Although anti cytokine therapies against IL-1, TNF-alpha or macrophage migration inhibitory factor showed promising results in experimental models of AP, the question remains as to whether similar antagonism during clinical pancreatitis would benefit patients with severe AP. Major considerations include the suitability of AP to cytokine antagonism in clinical settings, the possibility that such a therapy may lead to the development of immunosuppression and consequent infection, and whether a therapeutic window for such antagonism PMID- 15552898 TI - [Pancreatitis-associated gene mutations]. AB - Since the discovery of the cationic trypsinogen gene mutations in patients with hereditary pancreatitis, a variety of pancreatitis-associated gene mutations have been reported, including pancreatic secretory trypsin inhibitor and cystic fibrosis transmembrane conductance regulator. Although the patients with these mutations are rarely seen, genetic disorders inducing pancreatitis have provided us major breakthroughs to understand the molecular basis of the disease. Furthermore, the major stream in pancreatology has been evidenced in patients with hereditary pancreatitis: acute pancreatitis --> chronic pancreatitis --> pancreatic cancer. This report will focus on the pancreatitis-associated genes and the molecular mechanism of pancreatitis associating with these gene mutations. PMID- 15552899 TI - [Concept of SIRS and severe acute pancreatitis]. AB - Acute pancreatitis is a potentially fatal disease, the severity of which ranges from a mild edematous form to a severe necrotizing form. Most patients develop systemic inflammatory response syndrome (SIRS), which is induced by proinflammatory cytokines. The cytokine production can be induced by activation of Toll-like receptor. The breakdown products of the pancreatic and peripancreatic tissues by proteases might be the agonists. Cytokines are supposed to be produced as a biological defense system. However, cytokines may often evoke organ failure and/or immunosuppressive state, if they would be produced excessively. To express this complicated pathologic condition, Ogawa proposed a concept of LISIS(local inflammation-induced systemic immunosuppression syndrome). We have to pay attention to such aspects in the management of this disease. PMID- 15552900 TI - [Possible mechanisms of hepatitis C virus-related hepatocarcinogenesis]. AB - Persistent infection of hepatitis C virus (HCV) can lead to a high risk for hepatocellular carcinoma(HCC). HCV core protein plays important roles in HCV induced hepatocarcinogenesis, because its expression in mice causes hepatic steatosis and HCC without accompanying hepatitis. However, its precise mechanisms remain unclear. We investigated whether the HCV core protein alters the expression of the factors associated with hepatic steatosis and HCC in vivo. By Western immunoblot and Northern blot analyses, expression of the proteins including fatty acid-metabolizing enzymes and cell cycle regulators, which are induced by the activation of peroxisome proliferator-activated receptor alpha (PPARalpha), significantly increased in HCV core protein transgenic mice. This result suggests the possibility that PPARalpha activation might contribute to HCV core protein-mediated hepatocarcinogenesis. PMID- 15552902 TI - [Studies on allergic diseases: paradigm of management]. PMID- 15552901 TI - [Blood pressure variation and cardiovascular risk in hypertension]. AB - Disruption of circadian rhythm of blood pressure (BP) is associated with advanced target organ damage and poor cardiovascular prognosis. We studied silent cerebrovascular disease and stroke events in older Japanese patients with different nocturnal BP dipping. There was a J-shaped relationship of nocturnal dipping status with silent cerebral infarcts detected by brain MRI at baseline, and with stroke incidence during the follow-up period. The extreme-dippers (with marked nocturnal BP dipping) and risers (with higher nocturnal BP than awake BP) had a higher prevalence of silent cerebral infarcts and poorer stroke prognosis than those with appropriate nocturnal BP dipping (dippers). Extreme-dippers tended to have predominant systolic hypertension and increased BP variability. Several factors are affecting the diurnal BP variation pattern. The non-dipping pattern is associated with autonomic nervous dysfunction and poor sleep quality due to nocturnal behavior and sleep apnea. Extreme-dippers might have increased arterial stiffness with reduced circulating blood volume in addition to an excessive morning surge due to alpha-adrenergic hyperactivity. Morning BP surge, which is partly associated with nocturnal BP dipping status, was a predictor of stroke event independently for ambulatory BP level and silent cerebral infarcts. Antihypertensive medication that normalize the disrupted circadian BP variation might improve cardiovascular prognosis in high-risk hypertensive patients. PMID- 15552903 TI - [Diagnostic approach based on physiopathology. 1. Diagnosis of bronchial asthma]. PMID- 15552904 TI - [Diagnostic approach based on physiopathology. 2. Allergic rhinitis]. PMID- 15552905 TI - [Diagnostic approach based on physiopathology. 3. Atopic dermatitis]. PMID- 15552906 TI - [Diagnostic approach based on physiopathology. 4. Food allergy]. PMID- 15552907 TI - [Diagnostic approach based on physiopathology. 5. Drug hypersensitivity]. PMID- 15552908 TI - [Diagnostic tests and advances in their methods. 1. Skin reactions]. PMID- 15552909 TI - [Diagnostic tests and advances in their methods. 2. In vitro IgE antibody determination]. PMID- 15552910 TI - [Diagnostic tests and advances in their methods. 3. Histamine release test]. PMID- 15552911 TI - [Diagnostic tests and advances in their methods. 4. Induction of sputum production and ECP]. PMID- 15552912 TI - [Possibility for genetic diagnosis of allergic diseases]. PMID- 15552913 TI - [Quantitative analysis of environmental allergens and its significance]. PMID- 15552914 TI - [Quantification of the inflammatory mediators and their significance]. PMID- 15552915 TI - [Food-dependent exercise-induced anaphylaxis]. PMID- 15552916 TI - [Latex-fruit syndrome]. PMID- 15552917 TI - [Hypersensitivity to chemical substances]. PMID- 15552918 TI - [Guideline review. The 2003 guideline for prevention and management of asthma]. PMID- 15552919 TI - [Treatment of allergy within a framework of primary care: discussion]. PMID- 15552920 TI - [Cibenzoline-succinate-induced hypoglycemia]. PMID- 15552921 TI - [Nephrotic syndrome caused by focal segmental glomerulosclerosis with dentaterubral-pallidoluysian atrophy]. PMID- 15552922 TI - [Severe sleep apnea as a cause of hypertension]. PMID- 15552923 TI - [Allogeneic hematopoietic stem cell transplantation of myelodysplastic syndrome with pulmonary tuberculosis]. PMID- 15552924 TI - [Successful treatment of relapsed/refractory Hodgkin's lymphoma with tandem transplantation]. PMID- 15552925 TI - [Panhypopituitalism associated with HELLP syndrome]. PMID- 15552926 TI - [Antiphospholipid syndrome]. PMID- 15552928 TI - [Antibacterial agent-resistant organisms of the 21st century and their control]. PMID- 15552927 TI - [Prion disease: recent progress]. PMID- 15552929 TI - [Diseases of the domain of internal medicine and depression]. PMID- 15552930 TI - [Autoimmune pancreatitis]. PMID- 15552931 TI - [Effect of bcl-2 over-expression on the membrane structure of NG108-15 nerve cells: effectiveness of membrane-structure analysis using FACS for the first screening of apoptosis-inducing drugs]. AB - Bcl-2 is a Bcl-2 family protein that is known to be anti-apoptotic and is predominantly localized to the mitochondria. We previously showed that an analgesic, buprenorphine hydrochloride (Bph), induces apoptosis in the rodent derived nerve cell line, NG108-15, through the mitochondrial apoptotic route. A Bcl-2-overexpressing strain of NG108-15 cells, Bcl-2 (P2), was established, and the effect of Bcl-2 expression on Bph-induced apoptosis was compared between the mock vector-transfected NG108-15 cells and the Bcl-2 (P2) cells. The Bcl-2 (P2) cells died after treatment with Bph, and we observed all the biological and morphological markers of apoptosis that we tested for. In flow cytometric analysis, a difference in the cell membrane phospholipid flip-flop pattern-a feature of apoptosis- was observed between the NG108-15 cells and the Bcl-2 (P2) cells. Here, we show by flow cytometric analysis that Bcl-2 over-expression may affect the membrane structure of Bcl-2 (P2) cells. An increased fluorescein isothiocyanate (FITC) signal of annexin V-FITC, which typically represents phospholipid flip-flop of the cellular membrane in early apoptosis, was barely detected in the Bcl-2 (P2) cells. Since our previous study reported the localization of over-expressed Bcl-2 protein to the cell membrane of Bcl-2 (P2) cells, together these observations suggest that the Bcl-2 protein may affect the integrity of the structure of the NG108-15 cell membrane. PMID- 15552932 TI - [Urinary 8-hydroxy-2'-deoxyguanosin (8-OHdG) in patients with chronic liver diseases]. AB - Urinary 8-hydroxy-2'-deoxyguanosin(8-OHdG) has been reported as sensitive biomarker of oxidative DNA damage and also of oxidative stress. We measured the urinary 8-OHdG in patients with chronic liver diseases by competitive ELISA, and analyzed the relationship with clinical characteristics. Fifty patients (male/female: 22/28) with chronic liver disease were enrolled this study. The mean concentration of urinary 8-OHdG in healthy control and patients with liver cirrhosis, chronic hepatitis C, chronic hepatitis B, and autoimmune hepatitis were 10.40+/-3.14, 10.14+/-4.19, 11.79+/-5.58, 14.99+/-4.46, and 13.64+/-3.84 microg/gCr, respectively. There were no significant differences among the five group. The mean concentration of urinary 8-OHdG in inveterate drinker was significantly higher than that in non-drinker (16.67+/-4.29 vs. 11.19+/-4.80 microg/gCr, p<0.05). The smoking enhanced the elevation of urinary 8-OHdG in drinkers. In clinical characteristics, serum y-GTP, a marker of alcoholic liver disease, had significant positive correlation with urinary 8-OHdG on the drinker with chronic hepatitis. In addition, there was a positive correlation between serum ferritin levels and urinary 8-OHdG levels. Iron in the liver suggested oxidative damage of hepatocytes through the fenton reaction in patients with chronic liver disease. In conclusion, drinking and smoking induced liver damage by oxidative stress, and urinary 8-OHdG may be reliable marker of oxidative stress in patients with chronic liver disease. PMID- 15552933 TI - [Relevance of lipoprotein cholesterol levels measured by HPLC method to appearance midband on electrophoresis and remnant-like particle (RLP)-cholesterol levels]. AB - We have recently developed an HPLC method able to separate five lipoproteins (HDL, LDL, IDL, VLDL and chylomicron) followed by cholesterol measurement on each lipoprotein. As an application of this method, this study focused on analyses of triglyceride (TG)-rich lipoproteins, one of risk factor for atherosclerosis. The appearance of midband on electrophoresis is conceivably implicated in atherogenesis. The present study revealed that cholesterol levels in VLDL, IDL and LDL were significantly higher in midband-positive sera than negative sera. Cholesterol levels in remnant-like proteins, another atherogenic indicator, were significantly related to those in VLDL and Chylomicron measured by the present HPLC method. In conclusion, this novel HPLC method can provide valuable information for analyses on TG-rich lipoproteins. PMID- 15552934 TI - [Recent advances in immunologic laboratory testing for rheumatic diseases]. AB - Immunologic laboratory tests serve critical roles in the care of patients with various rheumatic diseases. These tests can provide relevant information of rheumatic diseases based on their immunopathophysiological condition. Although immunologic laboratory tests are quite useful for the determination of diagnosis and the estimation of disease activity, organ involvement and prognosis, they are frequently misused and resulted in an inappropriate diagnosis and treatment. Appropriate use of immunologic laboratory tests and accurate clinical interpretation of the test results can prevent false diagnosis and unnecessary treatment. In order to improve clinical care of patients with rheumatic diseases, clinicians caring patients with rheumatic disease should recognize meanings, characteristics and limitations of each result of immunologic laboratory testing. This article reviewed recent advances in immunologic laboratory testing such as antinuclear antibody, anti-DNA antibody and anti-neutrophil cytoplasmic antibody, and introduced guidelines for these testing. These guidelines based on evidences of EBM may contribute to the appropriate use of immunologic laboratory tests for rheumatic diseases. PMID- 15552935 TI - [Current topics on immunological laboratory tests-thyroid diseases]. AB - Current topics in the field of thyroid disease are the development of the second generation assay for TSH receptor antibody (TRAb) using recombinant human TSH receptor and the appearance of antineutrophil cytoplasmic antibodies(ANCA) in Graves' disease patients treated with propylthiouracil(PTU). This new TRAb assay is very useful, since the sensitivity and the specificity were almost 100%, respectively, in the diagnosis of Graves' disease. Furthermore, a new coated tube assay for the detection of blocking TRAb has been developed by using TSH/LH receptor chimera. The prevalence of ANCA is high in Graves' disease patients treated with PTU, but the clinical significance of ANCA is under controversy, since only a part of them develop vasculitis, and recently it has been reported that ANCA is frequently positive in Graves' disease patients before the onset of methimazole treatment. The 7th version of guidelines for the diagnosis of thyroid disease have been prepared by the Japan Thyroid Association, and opens to public inspection. They show the importance of immunological laboratory tests in this field. PMID- 15552936 TI - [Advances in neuroimmunological laboratory studies on neuromuscular diseases]. AB - The recent methodological advances in molecular biology, immunology, and genetics have clarified neuroimmunological problems in axonal Guillain-Barre syndrome, seronegative myasthenia gravis, paraneoplastic neurologic syndromes and many others. In addition to clinical and serological studies in peripheral neuropathies, the origins and measurement of anti-ganglioside antibodies and relationships to similar carbohydrate structures on infectious organisms are discussed in the context of molecular mimicry hypothesis, especially related with both the localization of relevant gangliosides in the nerve and clinical phenotypes. Major advances have been made in animal modeling of anti-ganglioside antibody-associated disease. An explanation for muscular weakness in 10-15% of patients with seronegative myasthenia gravis who lack autoantibodies to acetylcholine receptors(AchRs) appears to be the autoantibodies to muscle specific receptor tyrosine kinase (MuSK). MuSK mediates agrin-induced clustering of AchRs during synapse formation. These autoantibodies to the extracellular domain of MuSK inhibit its function in tissue culture. Isoelectric focusing (IEF) and agar gel electrophoresis (AGE) are used to examine cerebrospinal fluid (CSF) and sera from patients with multiple sclerosis (MS). The CSF oligoclonal IgG bands (OB) are less frequently observed in Japanese MS patients compared with Caucasian patients. Few optic-spinal form of MS (OS-MS) was positive for OB by agarose gel electrophoresis, but IEF is more sensitive than AGE. Recent IEF data revealed some OS-MS patients were positive for OB. The neuroimmunological advances revealed the relationship between the neuroimmunological diseases and the putative autoantibodies as diagnostic markers, for example, HAM and hnRNP-A1, MSand anti-hnRNP-B1 antibody, opsoclonus-myoclonus syndrome and anti-GluRdelta2 antibody, Rasmussen encephalitis and anti-GluR3 antibody, paraneoplastic brainstem encephalitis and anti-Ma2 antibody, and so on. PMID- 15552937 TI - [Transmission of tuberculosis (I)]. AB - BCG vaccination has been given to infants and children so extensively and repeatedly for more than 55 years in Japan that it is very difficult, or often impossible to diagnose tuberculosis infection by tuberculin testing. On the other hand, as the treatment of latent tuberculosis infection has become more and more important recently, diagnosis of infection is becoming more and more important at the occasions of contact survey. However, understanding of the health care workers about transmission of tuberculosis is incomplete frequently at present. This is the reason why the author has written this review. The author has described on the history of the progress of droplet nuclei infection theory, infectivity of tuberculosis by bacteriological status of the patients, importance of cough, susceptibility of the host, and environmental factor concerning transmission of tubercle bacilli in this issue. PMID- 15552938 TI - [Molecular epidemiological analysis by IS1245-based restriction fragment length polymorphism typing on cases with pulmonary Mycobacterium avium disease observed in the same family]. AB - INTRODUCTION: Mycobacterium avium-intracellulare complex (MAC) has become one of major human pathogens, however, its routes of transmission and environmental reservoirs causing human infection were not yet elucidated. We reported three families affected by pulmonary Mycobacterium avium (M. avium) disease. Previous reports on MAC diseases observed in the same family were very rare. The purposes of this study were to investigate whether the infected M. avium was the same strain among cases in the same family and to examine the possibility of human-to human transmission, or infection from exposure to a common environmental reservoir. METHODS: M. avium isolates from nine cases of three families were examined by DNA polymorphism based typing technique, restriction fragment length polymorphism (RFLP) analysis using insertion sequence IS1245 as a probe, to type the strains. Some isolates were subcultured to a single clone. RESULTS: All strains isolated from cases in the same family showed different patterns by the RFLP analysis. And not only simultaneous polyclonal infection but also repeated polyclonal infections were observed in some patients. DISCUSSION: The results suggest importance of underlying anti-mycobacterial immunological impairment and defects of local defense rather than virulence of infected strains as the pathogenesis of pulmonary M. avium disease. PMID- 15552939 TI - [Direct detection of rifampicin resistant Mycobacterium tuberculosis in sputum by line probe assay (LiPA)]. AB - OBJECTIVES: To examine the direct detection of rifampicin (RFP)-resistant Mycobacterium tuberculosis in sputum by Line Probe Assay (LiPA). MATERIALS AND METHODS: We collected 130 sputa and analyzed both by LiPA and the Amplicor M.tuberculosis assay. For culture-positive samples, RFP resistance testing was performed and compared with the results by LiPA. RESULTS: Eighty two out of 84 M. tuberculosis samples were detected by LiPA and all of 10 Mycobacteria other than M. tuberculosis (MOTT) samples and 36 negative samples were negative by LiPA. The detection rate is same as Amplicor. For culture-positive samples, LiPA showed mutation pattern for all of 22 RFP-resistant strains and wild type pattern for 19 of 20 RFP-sensitive strains. The one remaining showed mixed pattern of wild type and mutation pattern. CONCLUSION: The use of LiPA for sputum coould enable early detection of RFP-resistant tuberculosis and seems to be useful for the control of tuberculosis. PMID- 15552940 TI - [Case of pubic tuberculous osteomyelitis and pericarditis during anti tuberculosis chemotherapy]. AB - 56 year-old man was referred to the department of orthopedics in our hospital for further investigation on right inguinal pain. The patient was initially diagnosed as bacterial myelitis in right pubic bone and was treated with antibiotics. Since his symptom did not improve, the curettage was performed. Histological examination of the pubic bone obtained during the operation showed epithelioid cell granulomas with caseous necrosis, supporting the diagnosis of tuberculous osteomyelitis. Chest X-ray film revealed small nodular lesions in both upper lung fields. Sputum was positive for acid-fast bacilli and Amplified Mycobacterium Tuberculosis Direct Test (TB-MTD) was positive. The diagnosis of pubic tuberculous osteitis and pulmonary tuberculosis was confirmed. Specimens from the pubic bone and sputum were both culture positive for Mycobacterium tuberculosis, and bacilli were sensitive to anti-tuberculosis drugs. The antituberculosis chemotherapy was started with INH, RFP, SM and PZA. Symptoms had gradually improved, however 3 months after starting treatment, high fever developed and chest X-ray revealed heart enlargement and bilateral pleural effusion. Pericardial effusion showed exudative nature with lymphocyte predominancy and high level of ADA, 98.4 U/l. Pleural effusion was transudate. TB-MTD and culture were negative both in pericardial and pleural effusion. Paradoxical reaction was thought to be the cause of pericarditis. TB chemotherapy was continued and pericardial drainage was performed. One month later, fever improved, and pleural effusion and pericardial effusion disappeared. Transient elevated transaminase was observed, and was thought to be the side effect of anti TB drugs. All symptoms gradually improved, and he was discharged after 6 months TB chemotherapy. PMID- 15552941 TI - [An infant with tuberculosis who previously contacted an infant with congenital tuberculosis]. AB - We report a case of one-year and three-month old male infant with healed tuberculosis who was found by positive tuberculin skin test prior to scheduled BCG vaccination. Abdominal CT imaging revealed calcification in liver, spleen, and lymphnodes along pancreas and hepato-duodenal ligament, despite of normal chest imaging. A temporary intubated infant with congenital tuberculosis and/or his mother with untreated tuberculosis might infect him during his stay in the same NICU when he had digestive disease at three-month old. This report suggests that periodic contact investigation is needed for infants who had contact with the intubated infant with congenital tuberculosis in the same room, since cell mediated immunity is not well developed in infants. PMID- 15552942 TI - [Tuberculous infection and biological response in man]. AB - The characteristics and function of human lymphocytes in tuberculous morbid site were studied. Exudative-sensitized lymphocytes in tuberculous pleural fluid reacted to the specific antigen more effectively and produced higher titers of cytokines including interferon gamma (IFN-gamma) than circulating lymphocytes. CD4+/CD8- T-cell subset is responsible for the antigen-specific IFN-gamma production in pleural T lymphocytes of patients with tuberculous pleurisy. Thus, activated T lymphocytes concern the production of cytokines at the morbid site and they effectively exert local cellular immunity through the action of such cytokines. Immunofluorescence study showed increased production of inducible nitric oxide synthase (iNOS) and peroxynitrite in BCG-inoculated human alveolar macrophages (AM). Reverse transcriptase-polymerase chain reaction methods also revealed the higher expression of iNOS-coding mRNA. Colony assay demonstrated that human AM effectively killed BCG in their cytoplasm. However, treatment of AM with NG-monomethyl-L-arginine monoacetate resulted in markedly reduced killing activity. These results clearly show that BCG-induced NO and its reactive product with the oxygen radical, peroxynitrite, could play an important role in BCG killing in human AM. We measured the pleural concentrations of IFN-gamma, interferon-gamma-inducing cytokines; interleukin (IL)-12 and IL-18 and interferon gamma-inducible chemokines; IFN-gamma-inducible protein of 10 kDa (IP-10), monokine induced by IFN-gamma (Mig), and IFN-inducible T cell alpha chemoattractant (I-TAC). These cytokines and chemokines in tuberculous pleural effusions were much higher than those in malignant pleural effusions. These findings indicate that IFN-gamma plays an important role in the cell mediated immunity in tuberculosis. PMID- 15552943 TI - [Spinal cord protection during descending thoracic and thoracoabdominal aortic surgery]. AB - The incidence of postoperative paralysis after thoracic and thoracoabdominal aortic operations has decreased, but is still high in comparison with other operations. The analysis of the mechanism involved in the ischemic tolerance of the spinal cord could contribute to the protection of the spinal cord from ischemia. The identification of the Adamkiewicz artery and the predictive factors for postoperative paralysis in the preoperative period, the use of motor evoked potential, several adjuncts to keep the spinal cord circulation, the stabilization of the hemodynamics with good oxygenation, and hypothermia contribute to the prevention of the spinal cord ischemia. The anesthetics appropriate for the monitoring of the motor-evoked potential are propofol and fentanyl with or without ketamine. Among the anesthetic drugs, narcotics might exacerbate the motor function after the spinal cord ischemia. The analgesic drugs which do not aggravate the spinal cord dysfunction would be expected. Good cooperation of surgeons and anesthesiologists greatly contributes to the finding of the spinal cord ischemia during this operation. PMID- 15552944 TI - [The relationship between the respiratory parameters under spontaneous breathing and effect site propofol concentrations]. AB - BACKGROUND: The prediction of the hypnotic states is useful to maintain the adequate anesthesia. During propofol anesthesia, the respiratory depression has been documented in a dose-dependent manner. Therefore, we investigated whether the respiratory depression under the spontaneous breathing reflected the estimated effect site propofol concentrations (Cp) in a dose-dependent fashion. METHODS: We enrolled 12 patients for elective lower limb surgery under combined subarachnoid anesthesia and propofol sedation. The respiratory parameters and BIS were measured at the Cp of 5 microg x ml(-1) followed by the 0.5 microg x ml(-1) decrements until the patients' movement. Effective indices to predict patients' movement were determined by receiver-operator characteristics. RESULTS: The significant correlations within a particular patient between the respiratory parameters and Cp were observed, although those were not between the patients. An EtCO2 of 53 mmHg or greater represents a clinically determinant condition for non movement of the patients. CONCLUSIONS: We concluded that the respiratory parameters during spontaneous breathing were useful indices to predict the changes in the effect site propofol concentrations and to maintain the adequate anesthetic levels. PMID- 15552945 TI - [Patient dissatisfaction with anesthetic care]. AB - BACKGROUND: The evaluation of services by patients is an essential component of quality improvement in anesthesiology. Therefore, it is important to identify the factors for patient dissatisfaction. METHODS: We retrospectively studied 9974 consecutive patients who had received spinal or general anesthesia for elective surgery between 1999 and 2002. Pre-anesthetic, intra-anesthetic and post anesthetic variables were recorded and patient satisfaction was assessed using direct interviews at the post-anesthetic clinic. Qualitative data on dissatisfaction were obtained by asking patients' reasons for dissatisfaction. RESULTS: 348 of the 8843 respondents (3.9%) had dissatisfaction with anesthesia. The rates of dissatisfaction were higher in women than in men and in spinal anesthesia than in general anesthesia, and were observed mostly in the patients aged from 20 to 39 years. Qualitative data show that the common reasons for dissatisfaction with anesthesia were spinal anesthesia as the most dissatisfactory factor, followed by epidural anesthesia, postoperative pain, vomiting/nausea and memory of tracheal extubation. However, other various factors were associated with dissatisfaction. CONCLUSIONS: It is difficult for anesthesiologists to satisfy all patients, because patients' senses of values were varied. However, we conclude that anesthesiologists can improve the quality of anesthesia by enlightenment of the patient about anesthesia, and moreover, by better peri-anesthetic management for dissatisfactory factors with anesthesia. PMID- 15552946 TI - [Time course of thoracic epidural pressure and saphenous venous pressure changes with oxytocics after fetus delivery in parturients undergoing cesarean section]. AB - BACKGROUND: We examined the effects of oxytotics administered after fetus delivery on epidural and venous pressures in 40 parturients undergoing cesarean section under spinal anesthesia. METHODS: The subjects received 0.5% isobaric bupivacaine for spinal anesthesia. An epidural catheter placed at T11-12 and a venous catheter placed at left saphenous vein, were connected to pressure transducers to monitor each pressure. The subjects received intravenous methylergometrine 0.2 mg (methylergometrine group: n=20) or oxytocin 10 IU (oxytocin group: n=20) immediately after fetus delivery. Intramyometrial PGF2alpha was additionally administered from 5 minutes after fetus delivery in case of low uterine tone. Statistic analysis was performed with ANOVA. RESULTS: Epidural pressure in the methylergometrine group increased at placental delivery and the increase continued until 15 minutes after delivery (P<0.05). Epidural pressure in the oxytocin group increased at placental delivery and began to decrease at 5 minutes after delivery (P<0.05). Saphenous venous pressure began to increase at 10 minutes after delivery in the methylergometrine group (P<0.05). CONCLUSIONS: The increase in epidural pressure within 5 minutes after fetus delivery would be caused by uterine contraction in both groups. The vasoconstrictive effect in the methylergometrine group is likely to cause the epidural pressure increase after delivery. PMID- 15552947 TI - [Application of preoperative risk severity evaluation system (EuroSCORE=European system for cardiac operative risk evaluation) for cardiac operative patients]. AB - BACKGROUND: There is no widespread, well-accepted preoperative risk evaluation system for cardiac operative patients in Japan. We evaluated a scoring system for the prediction of postoperative morbidity and mortality in Japanese cardiac surgical patients. METHODS: Between August 1, 2002 and July 31, 2003 154 patients undergoing heart surgery with cardiopulmonary bypass were scored according to EuroSCORE (European system for cardiac operative risk evaluation). All patients were divided into three risk groups, low risk group (score 0-2), medium risk group (score 3-5), and high risk group (score 6 plus). We studied correlations between score points and postoperative morbidity and mortality. Data were analyzed using chi2 test, with a P-value of less than 0.05 as significant. RESULTS: The mean preoperative risk score was 5.6. The increase in preoperative risk score was associated with an increase in postoperative length of ventilation support and ICU stay. The low risk group had 35 patients with 0 death (0%), 5 morbidity (14.2%). The medium risk group had 57 patients with 2 death (3.5%), 14 morbidity (24.6%). The high risk group had 62 patients with 9 death (14.5%), 35 morbidity (56.5%). Overall, there were 11 death (7.1%), 54 morbidity (35.1%) in 154 patients. The risk score point correlated with postoperative morbidity and mortality. CONCLUSIONS: In spite of the limitation of total patient's number (N = 154) in this study, we recommend EuroSCORE as a simple and essential tool for the risk assesment of cardiac surgery and prediction of postoperative morbidity and mortality. PMID- 15552948 TI - [Laparoscopic ovarian cystectomy with abdominal wall lift during pregnancy under combined spinal-epidural anesthesia]. AB - Abdominal wall lift laparoscopic surgery is often used for patients during pregnancy because it is physiologically superior to CO2 pneumoperitoneum laparoscopic surgery. Operation for adnexal cysts is performed in the 1st trimester. We report seven cases of ovarian cysts during pregnancy, resected using gassless laparoscopic method with a whole abdominal wall lift under combined spinal-epidural anesthesia (CSEA). Combined spinal-epidural anesthesia had several advantages in these cases; 1. In the 1 st trimester, general anesthesia should be avoided. We could manage these cases without general anesthesia nor sedative medications. 2. During pregnancy, it is difficult to estimate the level of sensory blockade by spinal anesthesia. Epidural top-up helped us to easily control the level of sensory blockade. 3. Differential diagnosis of pain related to uterine contraction and postoperative pain is difficult. Post-operative analgesia was established by epidural PCA, thus anti uterine contraction medicines were prophylactically administered in only one of seven cases. There was no particular trouble during the anesthesia and all the operative procedures were performed uneventfully. Based on our limited experiences, CSEA may be a safe and appropriate anesthetic technique for laparoscopic ovarian cystectomy with abdominal wall lift during pregnancy. PMID- 15552949 TI - [Spinal anesthesia for 12 patients receiving home oxygen therapy]. AB - From Jan 2001 to Nov 2003, 12 patients receiving home oxygen therapy (HOT) underwent surgery under spinal anesthesia at Okayama Rosai Hospital. The basic diseases for HOT were emphysema (n=8), interstitial pneumonia (n=1), asthma (n=1) and lung cancer (n=1). Mean FEV1.0 and FVC were 0.85 l and 1.97 l, respectively. Mean PaO2 and PaCO2 were 76.5 mmHg and 45.5 mmHg, respectively, under nasal oxygen of 1.67 l x min(-1). Perioperative complications occured in 3 cases. In case 5, postoperative heart failure occured and was easily treated with diuretics. In case 8, intraoperative hypotension (systemic blood pressure less than 80 mmHg) occured. In case 12, the patient developed dyspnea because of high spinal anesthetic level of T1. She was not intubated because PaO2, PaCO2 and pH were not deteriorated. Perioperative PaO2, PaCO2 and pH were stable and there were no pulmonary or morbid complications in all cases. It is important to assess not only pulmonary function but also cardiovascular status by echocardiography and general physical status by Hugh-Jones classification in order to avoid severe complications. PMID- 15552950 TI - [A case of unexpected involuntary movement of lower extremities during epidural anesthesia]. AB - A 79-year-old man was scheduled for transurethral resection of the bladder tumor. Preoperative clinical and laboratory examinations revealed no abnormalities. An epidural catheter was inserted into the L3-4 interspace and 1.5% lidocaine 10 ml was injected smoothly with no abnormal signs. Thirty minutes after the injection, involuntary movement of lower extremities was noticed. When the catheter was removed after the operation, there was neither hemorrhage nor pain at the puncture site. Neither sensory disorder nor paralysis occurred while the movement continued for 150 minutes. No abnormal findings which might cause the involuntary movement were detected by clinical examinations including the cerebral CT and lumbar MRI. The detailed mechanism of the involuntary movement was not clarified, but exclusion of several complications including epidural hematoma by CT and MRI examination is essential for further diagnosis and treatment. PMID- 15552951 TI - [Anesthetic management of caesarean section in a patient with asymptomatic idiopathic prolonged QT interval syndrome]. AB - A 27-year-old patient with asymptomatic idiopathic prolonged QT interval syndrome underwent elective caesarean section. We selected combined spinal-epidural anesthesia to avoid excessive sympathetic stimulation. Bupivacaine 9 mg and lidocaine 100 mg were used for spinal anesthesia and epidural anesthesia, respectively. The anesthetic course was uneventful, and the infant showed no complication during the perioperative period. In conclusion, we consider combined spinal-epidural anesthesia as a safe anesthetic strategy for caesarean section in a patient with asymptomatic idiopathic prolonged QT interval syndrome. PMID- 15552952 TI - [Airway obstruction during one-lung ventilation using a bronchial blocker and a tracheostomy tube]. AB - We describe our experience with a 60-year-old man who had severe airway obstruction during one-lung ventilation with the tracheostomy tube using a bronchial blocker. The blocker, deriving from Univent tube, was passed through the tracheostomy tube and placed in the right main bronchus. We checked that the blocker was in appropriate place with a bronchofiberscope and obtained good one lung ventilation with the patient in the left lateral position. However, just after the start of operation, when the skin was incised, sever hypoxia and resultant bradycardia and hypotension occurred, probably because of not only malposition of blocker but also atelectasis in the upper lobe of the dependent lung by secretion. PMID- 15552953 TI - [A case of pharyngolaryngeal edema after posterior occipito-cervical operation]. AB - A 70-year-old female developed respiratory failure due to pharyngolaryngeal edema after posterior occipito-cervical fusion. She had a history of total thyroidectomy with bilateral neck dissection for advanced thyroid cancer associated with multiple lung metastases. However, her general condition was good, and she was not in cachexia. Her pulmonary function test revealed %VC of 54% and %FEV1.0 of 79%. This posterior occipito-cervical fusion was necessary for pain relief. Twenty-four hours after surgery she suddenly showed dyspnea, requiring tracheal intubation, and was supported by mechanical ventilation. The pharyngolaryngeal edema was recognized with bronchoscopy from the lower larynx with arytenoid region to nasal choana. In several days she recovered from pharyngolaryngeal edema and was extubated. She was discharged from ICU after close observation for 24 hours after extubation. She developed respiratory distress again due to difficulty in sputum expectoration, resulting in emergency tracheostomy an the floor at midnight. In this case, pharyngolaryngeal edema may have been caused by disturbance of lymphatic flow due to posterior occipito cervical fusion in a patient with a past history of bilateral neck dissection. We also reviewed the literature in this report. PMID- 15552954 TI - [Anesthetic management for non-cardiac operation with transesophageal echocardiography in a patient with left atrial myxoma]. AB - A 68-year-old female with left atrial myxoma underwent osteosynthesis of the night tibia. During the operation we observed the motion of the tumor continuously by transesophageal echocardiography (TEE). The operation was finished successfully without any cardiovascular complications. We think that TEE was useful for observation of left atrial myxoma during this non-cardiac surgery. PMID- 15552955 TI - [Unanticipated endotracheal tube displacement in a short-neck patient with a history of chronic rheumatoid arthritis: a comparison of three kinds of endotracheal tubes]. AB - Endotracheal tubes are known to have a risk of being displaced at neck extension or flexion or rotation. However, the displacement seldom causes clinical problems. An 74-year-old patient suffering from chronic rheumatoid arthritis underwent debridement in the infected knee under general anesthesia. Mechanical ventilation had to be continued because of poor oxygenation after the operation. She had been intubated with Hi Lo Evac endotracheal tube (HLE) for 3 days, then extubated. However, she again needed mechanical ventilation because of aggravated oxygenation. At that time, Profile cuff siliconised endotracheal tube (PCS) was placed for 3 days. We encountered dangerous displacement of HLE during the first mechanical ventilation, but did not have any clinically dangerous displacement of PCS during the second mechanical ventilation. Therefore, we compared the hardness of the three popular endotracheal tubes. We found the hardness of HLE was higher than the others. This might be one of the reasons for dangerous displacement of the endotracheal tube in our case. PMID- 15552956 TI - [A case of spontaneous intracranial hypotension effectively treated with cervical epidural blood patch]. AB - We report a case of cervical spontaneous intracranial hypotension (SIH). The patient is a 46-year-old woman with hard of hearing, dizziness and postural headache. Radionuclide cisternography (RNC) demonstrated a cerebrospinal fluid (CSF) leak at the low cervical region. Resolution of headache was obtained by conservative therapies of bed rest and intravenous (i.v.) drip infusion, but the dizziness remained. We performed epidural blood patch (EBP) with 8 ml of autologous blood at the C 6-7 interspace. Her dizziness disappeared after several days of EBP. SIH is an uncommon disease caused by CSF leakage. SIH is often self limiting, responding to bed rest and/or i.v. drip infusion. However, if the symptoms of SIH do not show a complete recovery by conservative therapies, EBP or continuous epidural infusion of saline has reportedly been used for the management of these cases. Most of the reported cases of EBP are in the thoracic and lumbar spinal regions. We have performed cervical EBP without complications, and the 6 month-follow-up MRI and RNC demonstrated that the abnormal findings had disappeared. PMID- 15552957 TI - [A survival case of cardiac arrest resuscitated initially by several anesthesiologists in an airport]. AB - We experienced a case of cardiac arrest in a man near the boarding gate area in an airport. An anesthesiologist witnessed the onset of his cardiac arrest. Three anesthesiologists undertook cardiopulmonary resuscitation until the arrival of the emergency medical service crew. An emergency medical kit equipped in an airplane could not be used during the resuscitation. Successful defibrillation was achieved by fourth 360 J shock performed by an emergency life-saving technician. The patient regained consciousness before hospitalization. Airport employees should be trained in techniques of basic life support in order to be able to resuscitate patients who have suffered cardiac arrest in an airport. Medical emergency kits equipped in airplanes should be made available for use in cases of cardiac arrest occurring near a boarding gate in an airport. PMID- 15552958 TI - [Minitracheotomy in treating upper airway obstruction after anterior cervical fusion]. AB - Respiratory disturbance occurs sometimes after anterior cervical fusion. This is often a result of upper airway obstruction due to prevertebral soft tissue swelling. Therefore we used cricothyrotomy tubes (Mini-Trach) in two patients with postoperative upper airway obstruction and performed assist-ventilation via the tubes. After starting to ventilate through Mini-Trach II, respiratory disturbance disappeared soon and respiration improved markedly. Mini-Trach II is an effective device to use in patients with post-operative respiratory disturbance due to prevertebral soft tissue swelling. PMID- 15552959 TI - [Development of rapid and sensitive screening methods for natural products using specific monoclonal antibodies]. PMID- 15552960 TI - Uterine cervical cancer: a holistic approach to mental health and it's socio psychological implications. AB - This correlation study investigated outpatients with early stage uterine cervical cancer. The subjects' mental health and its' relationship with demographic characteristics, clinical characteristics, and quality of life were examined. One hundred and seventy six patients from three major hospitals in the Fukuoka area were surveyed with a structured questionnaire. The status of mental health measured by CES-D (Center for Epidemiologic Studies Depression Scale) indicated an average score of 13+/-8 (mean+/-SD). No clinical parameters were found to have significant correlation to CES-D. However, increased pain (p< 0.001) and the absence of a husband or a partner (p < 0 .01) had greater CES-D score which indicated worse mental health outcome. The QOL (Quality of Life) scale developed for this study consists of the four domains: "Feel satisfied with life" (r = .526, p < 0. 01), "Find life worth living" (r= -.485, p < 0.01), "Feel no hindrance in daily life" (r= -.319, p<0.01), and "Feel no anxiety with illness" (r= -. 578, p < 0.01) all which have statistically significant correlations with CES-D scores respectively. Upon examination using the multi-regression model, a strong relationship between CES-D scores and "Feel no anxiety with illness" (r= .331, p<0.001) was evident. This showed to be the strongest indicator affecting the depression outcome, followed by "Strong pain" (r= .231, p<0.01). Clinical parameters, such as performance status, clinical stage, and medical treatment did not show any correlation to CES-D scores. The research suggests that the mental health of outpatients with uterine cervical cancer was influenced by pain and quality of life, rather than the clinical parameters. The presence of a husband or a partner played the role as social support to reduce the level of depression. In order to provide complete care of patients, pain management, anxiety management, and spousal involvement are crucial to patients' mental health, especially in the ongoing care of uterine cervical cancer. PMID- 15552961 TI - [Evaluation of prognosis and recurrence of hepatocellular carcinoma treated from 1988 to 2002 at Department of Medicine III, Kyushu University Hospital]. AB - From a series of 452 consecutive patients with hepatocellular carcinoma (HCC) admitted to the department of Medicine III, Kyushu University Hospital between January 1988 and December 2002, patients were evaluated, who met the following criteria: (a) the patients had not been diagnosed with HCC previously, (b) they had a solitary HCC less than 4 cm in diameter, or had fewer than three tumors with diameters less than 3 cm, and (c) patients with extrahepatic metastasis or vessel invasion were excluded. Five-year survival was more than 60%, and there were no significant differences in survival among the periods of 1988 - 1989, 1990 - 1994, and 1995 - 2002. There were significant differences in survival among patients with tumors of diameters < 2 cm (80%), 2 - 3 cm (60%), 3 < (40%) cm, and among those with tumor numbers of 1 (70%), 2 (50 %), 3 - 4 tumors (30%). The study also analyzed variables that could influence prognosis or recurrence of HCC, including tumor size and number, parameters of hepatic function, combined therapy with transcatheter arterial chemoembolization and local recurrence. Multivariate regression analysis showed that indocyanine green retention at 15 min serum albumin level, and local recurrence were significant independent predictors of survival, and that serum albumin and alanine aminotransferase levels were significant independent predictors of recurrence. In conclusion, achieving complete necrosis of HCC at first treatment to prevent local recurrence is important for improving the prognosis of patients with HCC. In addition, ameliorating hepatitis, by antiviral treatment for example, to maintain hepatic function is also important for improving both the prognosis and the prevention of the recurrence. PMID- 15552962 TI - [Progressive structural abnormalities in the brain of the schizophrenic]. AB - One hundred years ago, Kraepelin hypothesized that cerebral damage might cause dementia praecox. Many patients with schizophrenia show progressive clinical deterioration, although the evidence has remained unclear for the organic brain damage. Recently, brain abnormalities have been suggested, including enlargement of the lateral ventricles and reduction of the frontal and temporal lobes, due to the development of magnetic resonance imaging (MRI). Changes in brain structures have also been reported in the clinical course of schizophrenia. These structural brain studies have played important roles in clarifying the pathophysiology of schizophrenia and understanding the association of clinical symptoms with morphological brain change. However, there have been inconsistencies in the results on progressive structural changes. The selection of the patients, effects of medication, and social environmental factors may account for these inconsistencies. We have reviewed recent studies that found structural changes in the brain, including subcortical and ventricular of the schizophrenia. PMID- 15552963 TI - [The way to remission from chronic depression: a psychopathological consideration]. AB - In this report, we discuss the possibility of recovery from chronic depression from the psychopathologic viewpoint. In clinical practice, it is important to treat carefully the recovering process as well as not to dismiss the bipolarity, in order to prevent prolongation of the depression. We should consider the recovering process not as a mere interval preceding the remission, but an independent phase marked by a certain instability between two stable stages, severe depression and normal mental state. It is characterized by the psychopathology of the criticality, in which clinical meaning of a phenomenon could not determined univocally, varying from an extreme to another depending on therapeutic parameters. We propose some clinical devices to the chronic depression, defining its psychopathology as a diffuse dissemination of the criticality of the recovering process. As to general matter, it is necessary to switch therapeutic strategy for the acute stage to that proper to chronic period, maintaining the sick role on the patient. For the physiological treatment, stabilization of mood swing and regulation of circadian rhythm have priority over elevation of mood. Finally we discuss on the psychotherapeutic problems. On the one hand, chronic depressive patients have to liberate them from their narcissistic object relationship to establish autonomic ego. On the other hand, the therapist has to recognize that their depression has a certain truth on human being itself, to give them back the meaning of the disease. PMID- 15552964 TI - [Approach to depressogenic genes from genetic analyses of animal models]. AB - Human depression or mood disorder is defined as a complex disease, making positional cloning of susceptibility genes a formidable task. We have undertaken genetic analyses of three different animal models for depression, comparing our results with advanced database resources. We first performed quantitative trait loci (QTL) analysis on two mouse models of "despair", namely, the forced swim test (FST) and tail suspension test (TST), and detected multiple chromosomal loci that control immobility time in these tests. Since one QTL detected on mouse chromosome 11 harbors the GABA A receptor subunit genes, we tested these genes for association in human mood disorder patients. We obtained significant associations of the alpha 1 and alpha 6 subunit genes with the disease, particularly in females. This result was striking, because we had previously detected an epistatic interaction between mouse chromosomes 11 and X that regulates immobility time in these animals. Next, we performed genome-wide expression analyses using a rat model of depression, learned helplessness (LH). We found that in the frontal cortex of LH rats, a disease implicated region, the LIM kinase 1 gene (Limk 1) showed greatest alteration, in this case down regulation. By combining data from the QTL analysis of FST/TST and DNA microarray analysis of mouse frontal cortex, we identified adenylyl cyclase-associated CAP protein 1 (Cap 1) as another candidate gene for depression susceptibility. Both Limk 1 and Cap 1 are key players in the modulation of actin G-F conversion. In summary, our current study using animal models suggests disturbances of GABAergic neurotransmission and actin turnover as potential pathophysiologies for mood disorder. PMID- 15552965 TI - [Treatment standard for bipolar disorders]. AB - Lithium has clearly been the standard drug for treatment of acute mania, bipolar depression and for prophylaxis of manic and depressive phase. However, recent reviews show acute failure rates of lithium to be over 50%. It is well recognized mixed state, dysphoric mania or rapid cycler appears less likely to respond to lithium. It has also been clarified that in these subtypes of mania valproic acid and carbamazepine may be more efficacious than lithium. As for the treatment of bipolar depression, consensus seems to be emerging that combined treatment with a mood stabilizer and SSRI or SNRI is the first choice because TCA with a mood stabilizer increases the risk of switch to mania. PMID- 15552966 TI - Superior mediastinal and neck lymphatic mapping in mid- and lower-thoracic esophageal cancer as defined by ferumoxides-enhanced magnetic resonance imaging. AB - OBJECTIVE: The purpose of this study was to detect lymphatic drainage into the superior mediastinum and neck in thoracic esophageal cancer patients using ferumoxides-enhanced magnetic resonance imaging (MRI), and to have this information assist in determining the appropriate extent of lymphadenectomy. METHODS: Nine male patients with T2-T3 mid- and lower-thoracic esophageal cancer with lymph node metastasis were examined. The day before surgery, ferumoxides was endoscopically injected into the submucosal layer of the peritumoral lesion. Thereafter, lymph nodes in the superior mediastinum and neck, which were shown to be ferumoxides-enhanced on MRI, were harvested and evaluated; magnetic force from all harvested lymph nodes was measured ex vivo. RESULTS: MRI of the superior mediastinum and neck revealed 1(median) ferumoxides-enhanced lymph nodes in eight (89%) patients, and there was laterality in the lymphatic mapping in both areas. Of the 15 lymph nodes into which drainage was detected by enhanced MRI, 12 (80%) were magnetite-positive. In six patients (67%), magnetic resonance enhanced lymph nodes corresponded completely with the ex vivo magnetite examination, and in 3 patients (33%) there was partial agreement. In 3 (60%) of the 5 patients that showed paratracheal and/or supraclavicular lymph node metastases, all of the affected nodes were detected by MRI; in one patient some of the affected nodes were detected. CONCLUSION: Ferumoxides-enhanced MRI is useful for visualizing lymphatic drainage to the superior mediastinum and neck in thoracic esophageal cancer. It is an adequate procedure to form an estimate on the appropriate extent of lymphadenectomy. PMID- 15552967 TI - Granulocyte-colony stimulating factor enhanced the recruitment of bone marrow cells into the heart: time course evaluation of phenotypic differentiation in the doxorubicin-induced cardiomyopathic model. AB - OBJECTIVE: We traced and evaluated bone marrow-derived cells after granulocyte colony stimulating factor (G-CSF) treatment in the doxorubicin-induced cardiomyopathic heart in the time course. METHODS: C57BL/6 male mice received doxorubicin (15 mg/kg, i.p.). At 1 week after administration of doxorubicin, the mice were irradiated (900 cGy) followed by transplantation of bone marrow cells (BMT) derived from transgenic mice expressing green fluorescent protein (GFP) (1 x 10(6)) via a tail vein (BMT). G-group (n = 22) received G-CSF (50 microg/kg/day x 8 days, s.c.) after BMT, while C-group (n = 17) received saline. At 4 and 7 weeks after BMT, heart sections were fixed to evaluate bone marrow-derived GFP cells (BMD-GFP) with immunostaining for Troponin I (TnI), atrial-natriuretic peptide (ANP), connexin 43, von Willebrand factor, and Ki67. RESULT: There were migrated BMD-GFP in the whole heart of all animals. In the time course, migrated BMD-GFP increased in G-group. At 7 weeks the number of migrated BMD-GFP in G group (56.2 +/- 15.6/HPF) was larger than that in C-group (18.9 +/- 10.7/HPF) (p < 0.05). TnI- and connexin 43-positive BMD-GFP were spindle-shaped. Von Willebrand factor-positive BMD-GFP showed thinner-shape. ANP- and Ki67-positive BMD-GFP showed oval-shape. The numbers of these positive cells derived from BMD GFP, not different between the 2 groups, did not change from 4 to 7 weeks. CONCLUSION: The migration of BMD-GFP into the heart increased from 4 to 7 weeks after BMT by G-CSF. However, cardiomyocytes and endothelial cells originating from BMD-GFP were very few and neither increased nor changed in their shapes and numbers in the short term. PMID- 15552968 TI - Cosmetic approach for correction of simple congenital heart defects in female patients. AB - OBJECTIVE: Much more concern is recently paid to the cosmetics when simple congenital heart defect is repaired. Here we present the reversed U-shaped skin incision with lower partial sternotomy for juvenile or adult female patients. METHODS: Four patients with atrial septal defect (ASD) and one patient with ventricular septal defect underwent the operation with this method. Patients' ages ranged from 5 to 46 years old. RESULTS: Postoperative recovery in all cases was uneventful. Only the middle portion of the operative scar was visible when the breast was well developed. The scar was completely covered by the brassiere. A numbness of the upper medial portion of the left breast was recognized in one initial patient, who underwent ASD closure and concomitant right-side Maze procedure, necessitating a wider dissection than that used for other patients. CONCLUSION: A reversed U-shaped skin incision with a lower sternotomy could provide relatively comfortable operative fields for cardiac surgeons, and be cosmetically appealing to female patients who require a repair of simple congenital heart defect. PMID- 15552969 TI - Posterior mitral annuloplasty using autologous pericardium in the repair of posterior leaflet prolapse. AB - OBJECTIVE: Mitral valve repair is frequently performed now because it produces a favorable postoperative quality of life, as well as improved cardiac function. For the treatment of posterior leaflet prolapse, we perform a posterior mitral annuloplasty using an autologous pericardium. The present study assessed the efficacy of this operation. METHODS: From April 1999 to October 2003, 42 patients underwent a posterior mitral annuloplasty using autologous pericardium for the treatment of posterior leaflet prolapse. There were 15 men and 27 women with an average age of 63.9 +/- 11.8 years. The length of the autologous pericardium matched the length of the posterior leaflet annulus as measured with Carpentier Edwards ring sizer that was chosen based on the area of the anterior leaflet. RESULTS: The average size of the Carpentier-Edwards ring sizer that was used to determine the length of the autologous pericardium was 27.7 +/- 13 mm, and the absolute length of the pericardium was 50.9 +/- 1.8 mm, and the average intraoperative jet area, as assessed by transesophageal echocardiography, was 0.36 +/- 0.47 cm2. The five-year freedom from reoperation was 97.1%, while the freedom from significant residual mitral regurgitation (> or = 3+/4+) was 92.0%. Two patients (4.8%) developed systolic anterior motion, and one patient (2.4%) had a cerebral infarction. None of the patients died after surgery, and no patients developed complications such as hemolysis or ring detachment. CONCLUSIONS: Posterior mitral annuloplasty using an autologous pericardium was shown to be a superior technique because it allows a sufficient annular repair with no complications such as hemolysis or ring detachment. PMID- 15552970 TI - Subdural hematoma induced by cerebrospinal fluid drainage: potentially catastrophic complication of spinal cord protection after thoracoabdominal aneurysm repair. AB - A 50-year-old man underwent replacement of the descending thoracic aorta for a DeBakey type III-b chronic dissecting aortic aneurysm. During the surgery, lumbar cerebrospinal fluid (CSF) drainage with a 10 cm H2O pop-off pressure was used to protect against spinal cord ischemia. During cardiopulmonary bypass, the patient's pupils were isocoric, but anisocoric at end of the operation. As computed tomography of the brain showed right subdural hematoma, neurosurgical drainage was instituted emergently. Although some neurological deficit remained, the patient recovered well and was discharged. This case provides a strong reminder that CSF drainage for spinal cord protection against ischemia might induce subdural hematoma, which can be catastrophic during an operation for thoracoabdominal aortic aneurysm. PMID- 15552971 TI - Off-pump coronary artery bypass grafting in a patient with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria has not been described in patients undergoing off-pump coronary artery bypass grafting. A 65-year-old man who underwent percutaneous coronary stenting to the proximal left anterior descending artery for unstable angina was readmitted to our hospital complaining of recurrent chest pains. A coronary angiography revealed in-stent restenosis and new lesions of the distal left anterior descending artery as well as the left circumflex artery branch. He was found to have paroxysmal nocturnal hemoglobinuria which contributes to serious surgical complications including infection, bleeding, hemolysis and acute renal failure. After pancytopenia was treated with administration of granulocyte colony stimulating factor and transfusion of the washed red blood cells preoperatively, off-pump coronary artery bypass grafting was performed. Cardiopulmonary bypass was avoided in order to reduce activation of complements. His postoperative course was uneventful. Combination of appropriate perioperative management and off-pump cardiac surgery yielded an effective result in treating this patient without major complications. PMID- 15552972 TI - Off-pump quintuple coronary artery bypass grafting for situs inversus totalis. AB - Dextrocardia associated with situs inversus totalis is a rare condition. A 49 year-old man with dextrocardia and situs inversus totalis underwent off-pump quintuple coronary artery bypass grafting using bilateral internal thoracic arteries and bilateral radial arteries. The operative technique was similar to that of off-pump coronary artery bypass grafting for situs solitus. His postoperative recovery was uneventful. PMID- 15552973 TI - Lateral position prevents respiratory occlusion during surgical procedure under general anesthesia in the patient of huge anterior mediastinal lymphoblastic lymphoma. AB - Lymphoblastic lymphoma, an aggressive mediastinal mass, is recognized as serious threat to the patient in developing cardiac tamponade or airway obstruction. Surgical procedure is often required to relieve clinical emergency and to establish prompt pathological diagnosis. However, in such a patient, acute respiratory occlusion in the spine position can be a life-threatening complication during general anesthesia. We describe a 17-year-old man whose cardiac tamponade was treated by pericardial-pleural window through a left anterior thoracotomy in the lateral position. The patient recovered from hemodynamic compromise without showing respiratory occlusion during general anesthesia and remained in the lateral position until extubation. Pathological diagnosis was precursor T-lymphoblastic lymphoma. There were no complications attributable to the operative procedure. Further chemotherapy reduced the mediastinal mass in size after two weeks when the patient developed sepsis and died. Lateral position prevents respiratory occlusion during surgical procedure under general anesthesia in the patient of huge anterior mediastinal tumor with airway obstruction. PMID- 15552974 TI - Surgical management of tunnel-like subaortic stenosis via ventricular septal defect in a patient with the interrupted aortic arch. AB - A 24-day-old male with interrupted aortic arch (type B), ventricular septal defect, and tunnel-like subaortic stenosis underwent a one-stage surgical treatment. The operative procedure comprised reconstruction of the aortic arch, transatrial excision of the subaortic fibromuscular tissue via the ventricular sepatal defect, and patch closure of the defect. The patient tolerated the procedure well and the postoperative echocardiography demonstrated a residual pressure gradient across the left ventricular outflow tract of 20 mmHg. Our result suggests that the transatrial surgical management of subaortic stenosis via the ventricular sepatal defect produces a safe and promising surgical option. PMID- 15552975 TI - Large pulmonary arteriovenous malformation with hyperammonemia. AB - A 45-year-old female presented with generalized fatigue, unaccompanied by other symptoms. Investigation revealed severe anemia due to gastric bleeding, and hereditary hemorrhagic telangiectasia accompanied by a large pulmonary arteriovenous malformation (PAVM). Additionally, the presence of hepatic arteriovenous and portovenous shunts indicated hepatic involvement. In addition to hypoxemia due to right-to-left shunting in PAVM, hyperammonemia with normal hepatic function was detected. The large PAVM was successfully managed with surgical resection. Hyperammonemia, persisting despite the hemostasis of gastric bleeding, improved postoperatively in the absence of treatment directed at hepatic involvement. We believe that resection of large PAVM contributed to the improvement of hyperammonemia. PMID- 15552976 TI - Beating mitral valve replacement for a patient with porcelain aorta. AB - Beating mitral valve replacement was performed for mitral valve stenosis in a patient with a porcelain aorta after aortic valve replacement. The 77-year-old patient had developed heart failure several times. A chest computed tomographic scan revealed severe calcification on the ascending aorta and aortic arch. A cardiopulmonary bypass was established by bicaval drainage and aortic return after epiaortic echographic evaluation of the ascending aorta. The mitral valve was successfully replaced under a beating heart. PMID- 15552977 TI - [New perspectives on glycology for the progress of neurobiology]. PMID- 15552978 TI - [Challenge to a riddle of brain: neuroglycobiology in retrospect and anticipation]. PMID- 15552979 TI - [View of the Japanese biomedical research system]. PMID- 15552980 TI - [Japan consortium for glycobiology and glycotechnology; toward establishment of international network and systems glycobiology]. PMID- 15552981 TI - [Role of glycoconjugates in the developing Drosophila nervous system]. PMID- 15552982 TI - [Analysis of sugar codes in nervous system of the nematode C. elegans]. PMID- 15552983 TI - [Carbohydrate chains and a neuron specific ubiquitin ligase]. PMID- 15552984 TI - [Development and regeneration of the central nervous system and neural chondroitin sulfate proteoglycans]. PMID- 15552985 TI - [Mmorphogenesis of neurons and chondroitin sulfate]. PMID- 15552986 TI - [Bral1, Bral2: the novel brain specific-hyaluronan and protoglycan link protein genes]. PMID- 15552987 TI - [Retinal axon targeting and proteoglycans]. PMID- 15552988 TI - [Heparan sulfate, an essential of CNS development]. PMID- 15552989 TI - [Role of heparan sulfate endosulfatases in extracellular signal transduction]. PMID- 15552990 TI - [Role of glycolipids in myelin]. PMID- 15552991 TI - [Functional coupling between neuron and glia in neuronal sphingolipid biosynthesis]. PMID- 15552992 TI - [Lipid microdomains in nervous system]. PMID- 15552993 TI - [Gangliosides and synaptic function]. PMID- 15552994 TI - [Functional analysis of the nervous system in knockout mice of ganglioside synthase genes]. PMID- 15552995 TI - [Role of basigin, a glycoprotein belonging to the immunoglobulin superfamily, in the nervous system]. PMID- 15552996 TI - [Role of perlecan in neuromuscular function: studies in knockout mice and human disorders]. PMID- 15552997 TI - [Mice deficient in the HNK-1 carbohydrate exhibit impaired learning and memory]. PMID- 15552998 TI - [Function of dystroglycan in the nervous system]. PMID- 15552999 TI - [Abnormalities in myelinogenesis in glycolipid sulfotransferase-KO mice]. PMID- 15553000 TI - [Defective O-mannosyl glycosylation causes congenital muscular dystrophies]. PMID- 15553001 TI - [Fukuyama-type congenital muscular dystrophy and alpha-dystroglycanopathy]. PMID- 15553002 TI - [Pivotal role of ganglioside in aggregation of amyloid beta-protein]. PMID- 15553003 TI - [Glycosyltransferases as substrates for Alzheimer's beta-secretase]. PMID- 15553004 TI - [Abnormal connection of mutant PS1 to glycosphingolipids metabolism]. PMID- 15553005 TI - [Axonal Guillain-Barre syndrome and physiological role of gangliosides in saltatory conduction of myelinated nerve fibers]. PMID- 15553006 TI - [Miller Fisher syndrome]. PMID- 15553007 TI - [Differential sialylations on kindling model mouse]. PMID- 15553008 TI - Pulmonary lesions detected in population-based CT screening for lung cancer: reliable findings of benign lesions. AB - PURPOSE: To identify the characteristics of benign pulmonary lesions in order to reduce false-positive rates in screening computed tomography (CT) and in order to reduce frequency of follow-up high-resolution CT (HRCT). MATERIALS AND METHODS: We evaluated 238 screening-detected benign lesions and 23 screening-detected lung cancers for 12 characteristics: spiculation, well-defined margin, concave margin, polygonal shape, notch/lobulation, solid component, ground-glass opacity (GGO), air bronchogram, cavity, bubble-like appearance, pleural indentation, and vascular convergence. We also measured the lesion diameters to set a threshold for benign lesions. We tested combinations of these characteristics to differentiate benign lesions from lung cancers. RESULTS: By using certain combinations of the characteristics that showed statistically significant differences between benign lesions and lung cancers, benign lesions could be extracted without contamination by lung cancer in screening CT, when the combination included solid component as a positive finding. In HRCT, more than 80% of the benign lesions could be extracted without contamination by lung cancer when the combination included GGO as a negative finding. CONCLUSION: It seems possible to reduce the frequency of follow-up HRCT to establish a diagnosis of benign lesions using certain combinations of the characteristics of benign nodules. PMID- 15553009 TI - Ovarian vessel dilatation with ovarian mass on MR images: influence of pelvic venous plexus dilatation. AB - PURPOSE: We assessed the value of analyzing visualization of the ovarian vessels in relation to the features of ovarian masses using MR images. MATERIALS AND METHODS: 74 patients with 91 masses (61 benign lesions and 30 malignant lesions) were evaluated. Ovarian masses were classified using the following features: 1) benignancy, 2) characterization, and 3) maximum diameter. We assessed the relationship between visualization and the diameter of the ipsilateral ovarian vessels and the features of ovarian masses. In addition, we assessed the relationship between visualization and the diameter of ovarian vessels and pelvic venous plexus (PVP) dilatation. RESULTS: Regardless of tumor laterality, 82% of ovarian vessels were visualized (121/148 vessels) on MR images. There was a significant relationship between the average diameter of ovarian vessels and dilatation of PVP (p<0.001). In patients without dilatation, there was a stronger relationship between ovarian vessel average diameter and benignancy (p=0.007), and the character of ovarian mass (solid versus cystic; p<0.05, mixed versus cystic; p<0.01). CONCLUSION: We confirmed a strong relationship between the diameter of ovarian vessels and dilatation of PVP. If ovarian vessels are dilated in patients without dilatation of PVP, we could suggest the possibility of malignant ovarian tumor. PMID- 15553010 TI - Contrast enhancement for whole-body screening using multidetector row helical CT: comparison between uniphasic and biphasic injection protocols. AB - OBJECTIVE: To investigate which is more suitable for whole-body screening with multidetector row CT (MDCT) during one breathhold, a uniphasic or biphasic injection protocol for contrast material. SUBJECTS AND METHODS: Sixty patients received a volume of 1.7 mL x weight (kg) with iopamidol 300 mg iodine/mL. The patients were randomized into two injection protocols: A) a fixed injection rate of 2.0 mL/sec with a 70 sec delay, B) administration of 80% of the contrast material in 40 sec, then administration of the remaining 20% in 20 sec with an 80 sec delay. A helical scan from the apex of the lung to the base of the pelvic cavity was performed during one breathhold. CT attenuation values of the thoracic aorta, pulmonary artery, abdominal aorta, portal vein, superior vena cava (SVC), suprarenal and infrarenal inferior vena cava (IVC), liver, and pancreas were measured. Two radiologists visually assessed the degree enhancement of the IVC. In addition, the two radiologists visually assessed artifacts caused by contrast material in the subclavian vein and SVC using a four-point scale. RESULTS: Enhancement of the SVC in protocol A was significantly better than that in protocol B (p=0.04). Enhancement of the infrarenal IVC and liver in protocol B was significantly better than that in protocol A (p<0.01, p<0.01). Renal enhancement in protocol B was significantly better than that in protocol A (p=0.02). In all patients with both protocols A and B, enhancement of the suprarenal IVC was visually graded as acceptable or good. In all patients with protocol B, enhancement of the infrarenal IVC was graded as acceptable or good. In only 2/3 of patients with protocol A, enhancement of the infrarenal IVC was graded as acceptable or good. There was no significant difference in artifacts in the subclavian vein between the two protocols (p=0.77). Artifacts in the SVC in protocol B were significantly fewer than those in protocol A (p<0.01). CONCLUSION: Protocol B was more suitable for whole-body screening than protocol A, because of better enhancement of the liver and infrarenal IVC and fewer artifacts in the SVC. PMID- 15553011 TI - Peripheral vascular coil for peripheral MR angiography: phantom-based comparison with body coil by SNR, CNR, and visual evaluation. AB - AIM: To evaluate the image quality of MR angiography (MRA) with a peripheral vascular coil. MATERIALS AND METHODS: A peripheral vascular coil, a technical coil used in MRA of the pelvis and lower extremities, has 12 individual coil elements arranged in six pairs. We evaluated the performance of a peripheral vascular coil for image quality, signal-to-noise ratio (SNR), contrast-to-noise ratio (CNR), and visual evaluation by comparing it to a body coil using a phantom. RESULTS: SNR with the peripheral vascular coil was 1.5-2.2 times higher than that with the body coil in vertical distance, and 1.6-1.8 times higher in horizontal distance. CNR with the peripheral vascular coil was 2.1-3.8 times higher than that with the body coil. Visual evaluation with the peripheral vascular coil was 1.1-1.2 times higher than with the body coil in spin echo sequences, and 1.2-1.9 times higher in 3D fast spoiled GRASS (3D-FSPGR) sequences. CONCLUSION: The peripheral vascular coil for peripheral MRA is robust and accurate in evaluating peripheral vascular diseases. PMID- 15553012 TI - Impact of pretreatment cholinesterase level on survival of inoperable intrahepatic or hepatic-hilar carcinomas treated with three-dimensional conformal radiotherapy. AB - PURPOSE: Prognostic factors for inoperable hepatocellular carcinomas (HCC), intrahepatic cholangiocellular carcinomas (IHCC), and hepatic-hilar cholangiocellular carcinomas (HHCC) treated with three-dimensional conformal radiotherapy (3DCRT) were evaluated. MATERIALS AND METHODS: Thirty-five consecutive patients were treated with 3DCRT (HCC, 14; IHCC, 11; HHCC, 10). Doses of 3DCRT ranged from 30-70 Gy (mean, 51.5 Gy). The follow-up time of the 12 living patients was 5-39 (median, 11) months. RESULTS: Two-year overall survival rates of HCC, IHCC, and HHCC were 19%, 26%, and 39%, respectively. On univariate analysis, 3DCRT dose (<50 Gy vs. 50-54 Gy vs. > or =60 Gy) (p<0.01) and pre-3DCRT cholinesterase value (p<0.01) were statistically significant factors for survival. Two-year overall survival rates of the patients treated with <50 Gy, 50 54 Gy, and > or =60 Gy were 0%, 50%, and 17%, respectively. The difference in survival between patients given 50-54 Gy and > or =60 Gy was not statistically significant (p=0.13). Two-year overall survival rates of patients with higher and lower cholinesterase values were 43% and 0%, respectively. On multivariate analysis, the independent variable most predictive of survival was pre-3DCRT cholinesterase value (p<0.01). CONCLUSION: Pre-3DCRT cholinesterase value was a significant independent prognostic indicator for survival. Benefit of dose escalation above 60 Gy could not be demonstrated. PMID- 15553013 TI - Prediction of tumor control by tumor regression at 40 Gy/4 weeks of external beam irradiation for oropharyngeal carcinoma. AB - PURPOSE: To predict tumor control on the basis of tumor regression at 40 Gy/4 weeks of external beam radiotherapy (EBRT) for oropharyngeal carcinoma. METHODS: From 1990 to 1997, 55 patients (9 Stage I+II, 15 Stage III, and 31 Stage IV) with squamous cell carcinoma of the oropharynx were treated with curative EBRT (median dose, 66 Gy). Tumor response at 4 weeks was compared with loco-regional control. RESULTS: The five-year loco-regional control rate was 52%. Regression of primary tumors was more rapid and occurred at a rate higher than that of nodal lesions; however, nodal response served as a superior predictor of tumor control. Predictive factors for loco-regional control were tumor stage, loco-regional response at 4 weeks, and pathological differentiation. Stage III and non-well differentiated stage IV tumors demonstrating >50% regression at 4 weeks were controlled initially at 74% (17/23) and ultimately at 83% (19/23) by salvage therapy. In contrast, stage III+IV tumors displaying <50% regression and well differentiated stage IV tumors exhibiting >50% regression were controlled initially at 9% (2/23) and ultimately at 22% (5/23). CONCLUSION: Tumor regression at 40 Gy/4 weeks of EBRT is an important parameter in terms of prediction of loco regional control for stage III and IV disease. PMID- 15553014 TI - Evaluation of abdominal aortic aneurysm for endovascular stent-grafting with volume-rendered CT images of vessel lumen and thrombus. AB - PURPOSE: To evaluate the accuracy of CT images of abdominal aortic aneurysm (AAA) with volume-rendered (VR) display of vessel lumen and thrombus and to evaluate its usefulness in the planning of stent-grafting. MATERIALS AND METHODS: Helical CT was performed in 11 patients with AAA. Volume data of the vessel lumen and thrombus were separately extracted, and the VR images were reconstructed. Vessel measurements were made by five radiologists and compared with the axial and multiplanar reconstruction (MPR) images. Angiography and intravascular ultrasonography were used as standard references. The relations of aneurysmal sac to the major arterial branches and the subjective overall diagnostic value were evaluated by using a continuous rating scale. RESULTS: Accuracy of measurements was higher on VR images than on axial/MPR in 7 of 13 regions. Interobserver variance of VR images was smaller in 7 of 14 regions. Detection of renal and internal iliac artery involvements was better on VR images (p<0.05). Overall diagnostic value was also higher on VR images (p<0.05). CONCLUSION: VR CT images of vessel lumen and thrombus are supportive of and supplementary to conventional axial/MPR images in planning stent-grafting for AAA. PMID- 15553015 TI - Focal sparing of segment 2 in fatty liver: US appearance. AB - PURPOSE: To determine the frequency and sonographic features of focal sparing of fatty liver at segment 2. MATERIALS AND METHODS: The sonographic database was searched over the preceding three years. The criteria for sonographic diagnosis of focal sparing of fatty liver was an area of liver parenchyma of relatively low echogenicity with lack of mass effect. RESULTS: Six (0.9%) of 686 patients with diffuse fatty liver showed focal sparing at segment 2. The areas showed spherical or ovoid hypoechoic mass-like lesions measuring 2 to 6 cm in longest diameter (average, 3.8 cm). These all abutted on the posterior surface of segment 2 adjacent to the ligamentum venosum. These were segmental in two cases and subsegmental in four. CONCLUSION: In diffuse fatty liver, segment 2 is infrequently spared, and this may be a helpful adjunct sign of fatty liver. PMID- 15553016 TI - Multi-slice CT of thyroid nodules: comparison with ultrasonography. AB - PURPOSE: To evaluate the usefulness of multi-slice computed tomography (MSCT) in comparison with ultrasonography (US) for the differentiation of benign from malignant thyroid nodules and the evaluation of tumor extension. MATERIALS AND METHODS: Thirty patients with thyroid nodules (14 malignant, 16 benign) who underwent both MSCT and US participated in the present study. MSCT with contrast enhancement was performed, and 3D shaded volume rendering (SVR) and multiplanar reconstruction (MPR) were employed to differentiate benign from malignant nodules and to evaluate tumor extension. US images were obtained using a 7.5 MHz annular array probe. A three-point rating scale was used for image interpretation, and the kappa statistic was employed to evaluate agreement between MSCT and US. RESULTS: Sensitivity in differentiating benign from malignant nodules was found to be 85.7% for US and 78.6% for MSCT. Disagreement between MSCT and US occurred in assessing the homogeneity of the solid component and the presence of fibrous capsule. In two of seven T4 cases, MPR provided a more accurate diagnosis than US examination in detecting extracapsular invasion. CONCLUSION: For differential diagnosis of thyroid nodules, US was found to be better than MSCT. However, MSCT could be useful for the evaluation of advanced cases with suspicion of extracapsular extension. PMID- 15553017 TI - MRI of testicular epidermoid cyst. AB - This report presents and discusses the magnetic resonance imaging (MRI) and pathological findings of a case of testicular epidermoid cyst. A 35-year old man consulted a physician after he felt an enlargement of the right testis. Physical examination revealed a painless, non-tender, elastic hard mass in the right testis. A radiological examination was performed. MRI demonstrated an enlarged right testis, with a mass in it. The mass showed slightly low signal intensity on T1-weighted imaging, and high signal intensity on T2-weighted imaging. Both the T1- and T2-weighted images revealed a peripheral low signal intensity rim. The mass showed no enhancement. A right orchiectomy was carried out for epidermoid cyst. Pathologic examination showed a well-encapsulated nodule within the right testicular parenchyma, filled with a cheesy, yellow-white material similar in appearance to atheroma. The bull's-eye appearance is considered to depend on the presence of calcification. From the viewpoint of pathology, testicular epidermoid cysts do not always show bull's-eye. We then must recognize that some testicular tumors may be epidermoid cysts even without the bull's-eye findings. PMID- 15553018 TI - Benign pleomorphic adenoma with extensive cystic degeneration: unusual MR findings in two cases. AB - Two cases of benign pleomorphic adenoma (PA) with extensive cystic degeneration are described. In both cases magnetic resonance (MR) imaging revealed entirely cystic tumors with an irregularity at the cyst wall. Some additional findings were observed: one case showed layering within the cyst contents caused by intratumoral hemorrhage, and the other had a small mural nodule at the cyst wall that showed very low intensity on both T1- and T2-weighted images and corresponded pathologically to the hyaline stroma. In both cases, preoperative diagnosis was difficult based on the MR findings alone. Benign PA should be included in the differential diagnosis of an entirely cystic parotid mass. PMID- 15553019 TI - Retained lipiodized oil misdiagnosed as residual metallic material. AB - Retained foreign material is one of the most common issues in medical malpractice lawsuits. We report here a case in which an extremely high-density material was found within the pelvis, which proved to be harmless residual lipiodized oil. Although there have been reports of lipiodized oil being retained within the fallopian tube for more than 25 years, to our knowledge there have been no previous reports of lipiodized oil retention in areas other than the genital tract. PMID- 15553020 TI - Super-selective transcatheter arterial embolization of a traumatic splenic arteriovenous fistula. AB - We report a 61-year-old woman with arteriovenous fistula (AVF) of a splenic artery caused by traumatic injury, which was successfully occluded by percutaneous transcatheter arterial embolization (TAE). We embolized the AVF with a coaxial system using six microcoils. This case suggests that super-selective TAE for traumatic AVF of a splenic artery is an effective and primary choice of first treatment. The use of a microcatheter and microcoils is an alternative to surgery. PMID- 15553021 TI - [Beating heart coronary artery bypass grafting for acute myocardial infarction]. AB - We consider that off-pump coronary artery bypass grafting (CABG) [OPCAB], which results in local myocardial ischemia, is more effective for patients with acute myocardial infarction (AMI) than conventional CABG under cardiac arrest with global myocardial ischemia. Twenty-one patients (15 males, 6 females) received OPCAB for AMI, among whom surgery was performed following percutaneous coronary intervention (PCI) failure in 4 and PCI was performed prior to OPCAB in 2, while PCI was not performed in the remaining 15. Preoperatively, 16 patients had intraaortic balloon pumping (IABP), and 4 had IABP and percutaneous cardiopulmonary support (PCPS). The mean interval from onset to surgery was 11.7 (range 3 to 40) hours. In 20 cases, a complete revascularization was performed. The mean number of bypasses was 2.3 and OPCAB was carried out in 14 patients. In 2 cases, OPCAB was converted to on-pump beating CABG for complete revascularization. Fourteen patients (67%), each maintained with preoperative left ventricular ejection fraction (EF), were discharged with an elective bypass. Four patients died after on-pump beating CABG, in whom EF was lower than 10%. In addition, 3 died of low cardiac output syndrome (LOS) under PCPS and 1 of ventricular fibrillation. Based on our results, we considered that complete revascularization using OPCAB was effective for cases of AMI with PCI difficulty. However, in shock cases requiring PCPS, cardiac function was not improved even after revascularization. Therefore, it is necessary to study new procedures for shock cases during the period from onset to surgery. PMID- 15553022 TI - [Lobectomy for lung cancer in an elderly man on maintenance hemodialysis]. AB - An 82-year-old man who had been undergoing hemodialysis for chronic renal failure, 3 times a week for the last 3 years, was admitted to our hospital for the treatment of lung cancer. Under careful perioperative management, we successfully performed a lobectomy. Histological examination revealed well differentiated squamous cell carcinoma and the postoperative staging was pT1N0M0. On postoperative day 1, he was commenced on hemodialysis with nafamostat mesilate instead of heparin sodium, and there was no postoperative bleeding. The patient had an uneventful postoperative course and was discharged 3 weeks after his operation. PMID- 15553023 TI - [Ischemic left ventricular free wall rupture followed by ventricular septal perforation]. AB - A 78-year-old woman underwent coronary angiography because of acute onset of anterior chest pain, disclosing total occlusion of the left anterior descending artery. After this she fell into circulatory collapse. As a subsequent chest computed tomography (CT) revealed pericardial effusion, she was transferred to our hospital. At operation, an oozing lesion was found on the left ventricular anterior wall near the apex. Under extracorporeal membrane oxygenation, the bleeding was completely controlled by applying fibrin glue sheets. On the thirteenth day after operation, a new systolic murmur appeared with hemodynamic deterioration. Echocardiographic examination revealed ventricular septal perforation, and she underwent reoperation. The ventricular septal perforation was recognized on the apical anterior wall. It was repaired by an infarction exclusion method. The postoperative course was uneventful. Although ischemic ventricular double rupture is a very rare complication, patients who have risk factors for cardiac rupture need to be intensively followed up. PMID- 15553024 TI - [Thoracoscopic lung biopsy in the diagnosis of diffuse lung disease]. AB - The diagnosis of diffuse lung disease can only be made accurately by performing a thoracoscopic lung biopsy. We evaluated the safety and efficacy of thoracoscopic lung biopsy in diffuse lung disease. Between May 1999 and April 2003, 34 patients were referred to us for thoracoscopic lung biopsy. We compared the clinical characteristics, number of biopsies, operation times, postoperative course, complications, and mortality in 19 patients with dyspnea and 15 patients who were asymptomatic. Although the number of biopsies, operation times, duration of chest drainage, and postoperative stay did not differ significantly between the 2 groups, postoperative oxygen support was needed for significantly longer in the dyspnea group (p =0.038). A pathological diagnosis was established in all patients. Postoperative complications developed in 4 patients. A prolonged air leak in 1 patient and a wound infection in another resolved conservatively, but 2 patients died of acute exacerbation. Thus, we conclude that the indications for surgery must be carefully considered in dyspneic patients with diffuse lung to prevent acute exacerbation. PMID- 15553025 TI - [Change in the volume of extra cellular fluid in coronary artery bypass grafting (CABG); comparison between off-pump and on-pump CABG]. AB - During the perioperative period for off-pump coronary artery bypass surgery (OPCAB) and on-pump coronary artery bypass surgery (on-pump CABG), the volume of extra cellular fluid (ECF) was measured. The subjects were elective adult coronary artery bypass surgery cases, consisting of 13 OPCAB cases and 7 on-pump CABG cases. The ECF volume was measured the day before surgery, immediately after surgery, and 2, 4, 6, 8, 12, 24 and 48 hours after surgery, with a bioimpedance analyzer (XITRON 4000 C). ECF volume variation was defined as the difference from the preoperative value divided by body weight, and was expressed in %BW. At the same time, respiratory-index and leukocyte count were measured. The maximum postoperative ECF volume was 3.13 +/- 2.6 %BW in the OPCAB group and 5.36 +/- 2.0 %BW in the on-pump CABG group, that is, significantly higher in the on-pump CABG group. The ECF volume started to increase in the on-pump CABG group immediately after surgery (4.38 +/- 1.8 %BW in the on-pump CABG group and 2.07 +/- 2.4 %BW in the OPCAB group), reaching its peak 6 hours after surgery in the on-pump CABG group and 4 hours after surgery in the OPCAB group. Thereafter, the volume gradually decreased, and 48 hours after surgery the volume decreased in the OPCAB group to 0.064 +/- 1.5 %BW, or to about the same value as the preoperative value, whereas in the on-pump CABG group the volume remained high: 1.9 +/- 2.9 % BW. There was no significant difference between the 2 groups in the change in respiratory-index. The leukocyte count remained significantly higher in the on pump CABG group. The ECF volume was measured by the bioimpedance measuring method. This is a useful method of measuring the volume non-invasively and continuously. In the OPCAB group, the increase in postoperative ECF volume was less, and recovery to the preoperative level was faster than in the on-pump CABG group. PMID- 15553026 TI - [The efficacy of function water (electrolyzed strong acid solution) on open heart surgery; postoperative mediastinitis due to methicillin-resistant Staphylococcus aureus]. AB - Methicillin-resistant Staphylococcus aureus (MRSA) infection after cardiac surgery has recently increased. We compared the anti-inflammatory effect of an electrolyzed strong acid solution and a warm saline solution in patients with open heart surgery. These solutions were used for mediastinal irrigation before closing the sternum. Group A patients were irrigated by a warm saline solution, and group B patients were irrigated by an electrolyzed strong acid solution, administration of this water is safe, feasible, and easy for the prevention of MRSA infection. Postoperative infection was significantly decreased in the group B as compared in the group A. An electrolyzed strong acid solution may be effective on postoperative infection, particularly MRSA infection following open heart surgery. PMID- 15553027 TI - [Thoracoscopic surgery using a new bronchial blocker]. AB - We report use of a new bronchial blocker through a single-lumen endotracheal tube to achieve one-lung ventilation to perform thoracoscopic operation in patients in whom placement of the double-lumen tube failed and difficult intubation is predicted. The bronchial blocker tube was placed into the aimed bronchus under the bronchoscopic vision and the cuff of the blocker was inflated to achieve one lung ventilation. In all of the 4 patients, the bronchial blocker could be inserted and placed safely, quickly, and exactly under the fiberoptic flexible bronchoscopic vision to perform thoracoscopic operation without any complications. The new bronchial blocker tube through the indwelling endotracheal tube may have advantages in situations where placement of double-lumen endotracheal tubes is technically impossible or inappropriate. The use of the new bronchial blocker tube will, however, require careful evaluation in larger series. PMID- 15553028 TI - [Delayed surgical treatment of traumatic injury of the thoracic aorta]. AB - Two cases of traumatic aneurysm of the thoracic aorta associated with blunt chest trauma are described. We underwent the operation for the aorta at 23 days in case 1 and 39 days in case 2 after the injury. Immediate repair may be thought to be the first choice for acute traumatic injury of the thoracic aorta, but delayed surgery can be achieved more safely in the patients, especially with associated injury. PMID- 15553029 TI - [Lung cancer with ground glass opacity diagnosed by transbronchial lung biopsy using an ultrathin bronchoscope and virtual bronchoscopy]. AB - Although thoracic computed tomography (CT) screening indicated that there are many patients who have pulmonary shadow with ground glass opacity, it is sometimes difficult to obtain the appropriate specimens for histological diagnosis of such patients. We herein report a lung cancer patient with ground glass opacity who was diagnosed preoperatively by an ultrathin bronchoscope and virtual bronchoscopy. A 78-year-old female was admitted to our hospital due to bacterial pneumonia. At the admission, CT showed another abnormal small shadow in her right middle lobe. Since the shadow was not visible by fluoroscopy, we reconstructed the images of virtual bronchoscopy using the data obtained by multidetector CT. The location of the shadow was determined in the peripheral area of a dorsal branch of right B4aialpha. Then the transbronchial lung biopsy using an ultrathin bronchoscope with simultaneous CT guidance was performed. The histological findings of the biopsy specimens revealed that the shadow was highly suspicious for malignancy. Therefore, the right middle lobectomy was conducted, and the tumor was diagnosed as an adenocarcinoma. An ultrathin bronchoscope with virtual bronchoscopy is useful to diagnose a pulmonary shadow with ground glass opacity. PMID- 15553030 TI - [Adult mitral and tricuspid valve regurgitation due to patent ductus arteriosus combined with atrial fibrillation; report of a case]. AB - 73-year-old female with mitral and tricuspid valve regurgitation due to patent ductus arteriosus (PDA) with atrial fibrillation (AF) was reported. The patient was admitted for dyspnea on effort. She was diagnosed as mitral and tricuspid valve regurgitation due to PDA with AF by transthoracic echocardiography. She underwent mitral valve plasty (quadrangular resection of prolapsed posterior leaflet and annuloplasty with Duran flexible-ring 25 mm), tricuspid annuloplasty (De Vega annuloplasty), PDA direct closure and radiofrequency modified maze procedure. Postoperative echocardiogram showed good mitral and tricuspid valve function. He recovered well after the operation and was discharged in stable sinus rhythm. PMID- 15553031 TI - [Primary cardiac malignant lymphoma diagnosed intraoperatively during aortic valve repair; report of a case]. AB - A 58-year-old male who complained of chest oppression on effort was diagnosed as aortic regurgitation by echocardiography. The cardiac catheterization was showed moderate aortic regurgitation. He was referred to our hospital for aortic valve replacement. We considered preoperatively that the aortic regurgitation is due to infective endocarditis or aortitis syndrome. We have not noticed cardiac tumor at all until aortic valve replacement. Postoperative pathological diagnosis of the tumor was malignant lymphoma. The tumor had a characteristic architecture of a cardiac MICE (mesothelial/monocytic incidental cardiac excrescence). After the operation, additional chemotherapy using doxorubicin and cysplatin was performed. The patient has been doing well for 5 years after operation. PMID- 15553032 TI - [Congenital supravalvular aortic stenosis; report of a case]. AB - We experienced a successful surgical case of extended aortoplasty by means of Doty's method (two sinus reconstruction) for congenital supravalvular aortic stenosis (SAS). Case was 12-year-old boy, who had no complaint except heart murmur. The retrograde aortography demonstrated localized stenosis just above the aortic valve, and it was an hour-glass type. The preoperative peak systolic pressure gradient between the left ventricle and ascending aorta was 56 mmHg, which was improved postoperatively. This case showed excellent results. Doty's aortoplasty was favorable method for SAS without deformity of aortic valve and coronary obstruction. PMID- 15553033 TI - [Aortic valve papillary fibroelastoma found at reoperation after open mitral commissurotomy; report of a case]. AB - In 1982, a 63-year-old woman underwent open mitral commissurotomy for rheumatic mitral stenosis. She was referred to our hospital for surgical treatment of mitral restenosis in 2003. Preoperative echocardiography showed rheumatic mitral valve restenosis and demonstrated a mobile mass of the aortic valve. During the operation, a mitral valve replacement was performed using prosthetic valve (SJM 29 mm) and a tricuspid valve was repaired by annuroplasty (De Vega). Through the aortotomy, a multiple frond-like tumor, measuring 5 mm, was found attached to the free-edge of the left-coronary cusp of the aortic valve. The resection of the tumor was performed without valve replacement. The histopathologic diagnosis was papillary fibroelastoma. The postoperative course was uneventful. In the pathogenesis of this case, the iatrogenic factor may have played a role in tumor development. PMID- 15553034 TI - [Mitral valve plasty and coronary artery bypass grafting under beating heart cardiopulmonary bypass for severe left ventricular dysfunction; report of a case]. AB - A 54-year-old man with ischemic mitral regurgitation and severe heart failure due to broad myocardial infarction successfully underwent mitral valve plasty and coronary artery bypass grafting under beating heart. He had an old anterior myocardial infarction and was admitted to our hospital with acute inferior myocardial infarction. Two weeks later, the cathetelization revealed moderate mitral regurgitation and triple vessel coronary artery disease. We selected antegrade continuous blood perfusion for myocardial protection on operation. He recovered uneventfully and discharged on postoperative day 31. We could perform this procedure safety and satisfactorily, we could this procedure for heart valve operation with other complications. PMID- 15553035 TI - [Posterior mediastinal paraganglioma with a 60-year history; report of a case]. AB - We report a case of a 68-year-old man with a posterior mediastinal tumor. He remembered having difficulty in food passage through his esophagus in childhood. The preoperative thoracic computed tomography (CT) revealed a 5 x 3 cm mass with strong enhancement in the posterior mediastinum. A right thoracotomy was performed to resect this indeterminate mass. Complete resection was achieved. The pathologic diagnosis was paraganglioma. Postoperatively, he experienced more comfortable food passage through his esophagus than ever before. There has been no sign of recurrence 3 years after the operation. The tumor was clinically diagnosed as benign paraganglioma because of its long silent course. PMID- 15553036 TI - [Simultaneous spontaneous pneumomediastinum caused by vocal exercise; report of 2 cases]. AB - Two cases of spontaneous pneumomediastinum caused by vocal exercise were reported. Two 18-year-old men were admitted to our hospital simultaneously in April 2003, because of cervical discomfort and chest pain after vocal exercise of self introduction as a event of freshman in their college. Their chest X-ray film and chest computed tomography (CT) demonstrated typical pneumomediastinum. One of them also showed thoracic epidural emphysema without any particular neurological deficit. Both cases completely recovered by only conservative therapy within 5 days. Spontaneous pneumomediastinum which occurs frequently in young men is thought to be a relatively rare disease showing a good prognosis. It seems important to consider this rare condition when the young man complaints chest pain and discomfort around their neck. We thought that there must be a high possibility of this disease being overlooked as a mere chest pain in young man of unknown reason until now. It is our conclusion that spontaneous pneumomediastinum is a really benign condition that requires no specific examination nor therapy. PMID- 15553037 TI - [Pulmonary embolectomy followed by lobectomy to treat for massive endobronchial bleeding; report of a case]. AB - We describe a case of successful pulmonary embolectomy that required lobectomy to treat for endobronchial bleeding. Sixty-nine-year-old female got into shock required CPR during rehabilitation after replacement of femur head. Her enhanced computed tomography (CT) showed massive embolism in pulmonary artery. Emergency embolectomy was performed under cardiopulmonary bypass (CPB). A videoscope was employed to see inside the pulmonary artery. At the end of the CPB, endobronchial bleeding occurred. The right bronchus was occluded by balloon and CPB was weaned. The right lower lobectomy was done on the next day. She had got pyothorax by methicillin-resistant Staphylococcus aureus (MRSA), and has recovered after intravenous administration of antibiotics and continuous wash out of pleural space by saline containing 0.02% povidon-iodine. Postoperative scan showed no defects of blood flow and ventilation except resected lobe. PMID- 15553038 TI - [A large nontuberculous mycobacterial granuloma mimicking lung cancer; report of a case]. AB - A 45-year-old woman with no immunodeficiency or clinical symptoms presented to our hospital for treatment of an enlarging spherical lung tumor in 4.0-cm diameter. Chest radiography 8 years ago had shown a 1.5-cm-diameter tumor. Chest computed tomography (CT) showed the solitary tumor, located in the right apical segment, to have an irregular margin without satellite lesions or cavitations. Mycobacterium avium complex (MAC) was cultured in tumor specimens incised during transbronchial biopsy. Right upper lobectomy was performed because of resistance to 6-month antituberculosis treatment. Pathological findings showed a MAC infected granuloma with caseous necrosis. Postoperative course was uneventful and she had no recurrence 3 years after surgery. These findings suggest that nontuberculous mycobacterial granuloma can enlarge without clinical manifestations or any satellite lesions and cavitations, leading to a misdiagnosis of lung cancer. PMID- 15553039 TI - [Bronchioloalveolar carcinoma with acute respiratory failure due to hemoptysis; report of a case]. AB - A 69-year-old male was admitted to our hospital for hemoptysis and dyspnea. Because of his deteriorating respiratory distress, he was intubated and controlled by respirator for 3 days. He was diagnosed with adenocarcinoma of the lung by the sputum examination and chest computed tomography (CT) revealed an infiltration shadow in the peripheral superior ventral segment (S3) of the right upper lobe. He underwent right upper lobectomy with video-assisted thoracic surgery. Microscopic findings of the resected specimen measuring 10 x 10 x 7 cm revealed mucin-producing bronchioloalveolar carcinoma (BAC) with metastases in lymph nodes and the same lobe (S2b). We reported a rare case of BAC with hemoptysis. PMID- 15553040 TI - [Recurrent breast cancer to the sternum 15 years after radical mastectomy and primary lung cancer; report of a case]. AB - We reported a successful operative case of solitary metastasis in the sternum 15 years after radical operation for a breast cancer and a primary lung cancer. The patient was a 59-year-old woman who was admitted for skin ulcer and tumor of the anterior chest wall. Histological diagnosis by skin biopsy was metastasis of breast cancer. Concurrently, chest computed tomography (CT) revealed a coin lesion with slight spiculation at the right lower lobe. Because histological diagnosis by the partial resection of the right lower lobe was primary lung cancer, we performed right lower lobectomy. Twenty-four days after the operation, subtotal sternal resection was carried out. She is alive and well without any complaints. PMID- 15553041 TI - [Anemia in Japanese women: the current situation and the cause]. PMID- 15553042 TI - [Analysis of varicella zoster virus infection following allogeneic stem cell transplants]. AB - The purpose of this study was to evaluate patients who contracted the varicella zoster virus infection (VZV) following their allogeneic stem cell transplants. We retrospectively reviewed the incidence and the timing of varicella zoster virus (VZV) infections, including the clinical course, complications, and associated clinical risk factors. Between January 1998 and April 2003, a total of 71 patients received allogeneic stem cell transplants in our hospital. For prophylaxis of the herpes virus infection, all patients were given a daily oral 1000 mg dose of acyclovir from day -7 to day +35. Among the 71 patients, 28 of them (39.4%) developed VZV infection between day 77 and day 980 (median 182 days) following their allogeneic stem cell transplants. In 21 of these infected patients (75%) the occurrence was within the first 300 days after the transplant. Twenty-two patients (78.5%) were under treatment with immunosuppressive agents. Twenty-six patients developed only one episode of the VZV infection after their transplants, but two other patients developed two episodes. Twenty one patients (75%) stricken with the VZV infections had cutaneous reactivation infections of a single dermatome, and in one patient two dermatomes were affected. Five patients (17.8%) developed disseminated cutaneous zoster, and one patient (3.6%) developed a visceral infection. Treatment with acyclovir (oral or drip infusion) was successful in 25 patients. Two patients improved with vidarabine treatment, however the patient with the visceral infection died despite the use of acyclovir. The incidence of visceral infection was low, but the one case was fatal. PMID- 15553043 TI - [Splenic marginal zone lymphoma associated with antiphospholipid antibodies]. AB - A 61-year-old woman experienced a high fever with anemia and APTT prolongation after suffering a herpes zoster virus infection. Physical examination revealed a large splenomegaly without lymphadenopathy. Laboratory evaluations were positive for lupus anticoagulant (LA) and monoclonal IgM-kappa protein. LA was associated with the presence of anti-beta2GPI antibody, anti-cardiolipin antibody, and anti prothrombin antibody. Moreover, the results of factors IX, XI, and XII assays and CRP and FDP-E were disturbed. A splenectomy was performed, and a splenic marginal zone lymphoma (SMZL) was diagnosed. All hematological findings rapidly recovered after the splenectomy. No thrombotic events occurred after the splenectomy even though thrombosis prophylaxis was not performed. The clinical course suggested that the SMZL-producing antibody induced immunological abnormalities in the labolatory tests. Since the patient suffered disease progression soon after the splenectomy, an autologous peripheral stem cell transplantation with rituximab administration was performed. PMID- 15553044 TI - [Methotrexate-induced interstitial pneumonitis in a child with acute lymphoblastic leukemia]. AB - We report a case of 5-year-old boy with acute lymphoblastic leukemia who developed interstitial pneumonitis induced by methotrexate (MTX). The patient was hospitalized with fever, cough, dyspnea and hypoxemia during maintenance treatment with low dose MTX and 6-mercaptopurine. A diagnosis of MTX pneumonitis was made based on the clinical findings, viral and serologic studies, negative microbiology and the radiological features. The patient recovered after cessation of the MTX treatment. Interstitial pneumonitis caused by MTX is well-recognized and the prevalence has been estimated to be 0.3-7.5% among patients with adult rheumatoid arthritis. However, there are few reports in the literature regarding this adverse effect in patients with leukemia. Furthermore, very few cases of childhood leukemia have been reported regarding MTX induced interstitial pneumonitis. Physicians should be aware of this rare complication during maintenance treatment with weekly low dose MTX for acute lymphoblastic leukemia in children. PMID- 15553045 TI - [Imatinib mesylate plus G-CSF therapy for chronic myelogenous leukemia in the blastic crisis]. AB - Imatinib mesylate (imatinib) has shown significant effects in patients with chronic myelogenous leukemia. However, hematological toxicity often occurs and requires dosage reduction or discontinuation of imatinib treatment. A patient with chronic myelogenous leukemia in the blastic crisis received granulocyte colony stimulating factor (G-CSF) simultaneously with imatinib. The patient was continuously treated with imatinib and G-CSF and achieved remission without any severe infection or neutropenia. There are a few reports on the efficacy of combined therapy with G-CSF and imatinib; however, the results in our case are rare suggesting that the use of G-CSF is effective for preventing severe infection. G-CSF enables continuous treatment with high-dose imatinib. PMID- 15553046 TI - [Successful treatment of POEMS syndrome with high-dose chemotherapy and autologous peripheral blood stem cell transplantation]. AB - A 42-year-old woman presented with pericardial and pleural effusion, ascites and para-aortic lymphadenopathy of unknown etiology. Six months later she was admitted with fever, pain and motor disturbance of lower limbs, and exacerbation of the effusion, ascites and edema. Physical examination showed hepatosplenomegaly, skin pigmentation and hypertrichosis. Immunoelectophoresis revealed monoclonal IgA-lambda protein in the serum and Bence-Jones protein lambda in the urine. Bone marrow aspiration showed a mild increase of atypical plasma cells. Vascular endothelial growth factor (VEGF) had markedly increased to 10,900 pg/ml. Electromyography showed changes suggestive of demyelination. These clinical features were consistent with the diagnosis of POEMS syndrome. VAD chemotherapy was not effective for the effusion and neuropathic deterioration. After control of the massive pleural effusion by chest tube drainage, peripheral blood stem cell (PBSC) collection was performed with cyclophosphamide and G-CSF. The patient received melphalan 100 mg/m2 on 2 consecutive days and the PBSC were infused 2 days later. The bone marrow recovered rapidly and the pericardial and pleural effusion disappeared completely. Her performance status markedly improved from a bedridden state. High-dose melphalan with auto-PBSCT should be investigated further as a recommended therapy for POEMS syndrome. PMID- 15553047 TI - [Hypercholesterolemia as a part of chronic GVHD after allogeneic stem cell transplantation]. AB - A 45-year-old female with acute myelogenous leukemia (AML-M6) received an allogeneic stem cell transplantation from an HLA-identical sibling donor in June 2002. Prophylaxis against graft-versus-host disease (GVHD) consisted of cyclosporine (CsA) and short-term methotrexate. Acute GVHD did not occur and CsA was discontinued on day 145 after transplantation. However, soon thereafter she suffered from conjunctivitis, stomatitis and liver dysfunction with hypercholesterolemia and was diagnosed as having chronic GVHD. The liver dysfunction and hypercholesterolemia failed to improve despite the administration of CsA and prednisolone. Atrovastatin was not effective and immunosuppressive therapy for two months including ursodeoxycholic acid finally improved the jaundice and hypercholesterolemia. Although lipid metabolism analysis in this case disclosed the same findings as in other intrahepatic cholestatic liver diseases, the results show that the improvement of hypercholesterolemia in chronic GVHD needs the same treatment as chronic GVHD. PMID- 15553048 TI - [CD56-positive peripheral T-cell lymphoma primarily presenting with tonsillar swelling]. AB - A 51-year-old woman was admitted to our hospital with tonsillar swelling. After tonsillectomy was performed, she was diagnosed as having CD56-positive T-cell lymphoma, mainly composed of small and medium-sized atypical cells. An immunohistochemical study showed that the malignant lymphocytes were positive for CD3, CD8, CD56, TIA-1 and granzyme B, while negative for CD20, CD5 and CD10. Flowcytometry demonstrated the lymphocytes were positive for CD56. Southern blot analysis revealed a rearrangement of the T-cell receptor gamma chain. The disease stage by Ann Arbor staging classification was II B. We provided MCEC therapy followed by autologous peripheral blood stem cell transplantation, and complete remission (CR) was achieved. Two months after CR, however, the patient relapsed with peritonitis due to perforation of an ileal tumor, and died of sepsis. It is rare for CD56-positive T-cell lymphoma to occur primarily in the tonsils. Because small bowel ulcers were revealed during the course of induction chemotherapy, we report a valuable case in which suspected CD56-positive enteropathy-type T-cell lymphoma (ETL) occurred primarily in the tonsils. PMID- 15553049 TI - [Multiple myeloma of the IgD-lambda type invading CNS]. AB - A 52-year-old woman was admitted to the gynecological department of our hospital on July 29, 2002 because of a right lower abdominal mass. She has been suffering from pain in the right leg and inguinal area for a month before coming to the hospital. She was found to have pancytopenia and high serum levels of LDH and IgD. A bone marrow examination showed 63.8% of plasma cells and serum immunoelectrophoresis showed M-protein of the IgD-lambda type. She was diagnosed as having multiple myeloma and transferred to our department. VAD therapy was started from August 22. Although the plasma cells in the bone marrow almost disappeared, the right lower abdominal mass remained and a new mass appeared on the right frontal chest wall after two courses of the treatment. Combination chemotherapy with vincristine, ranimustine, melphalan, and dexamethasone (ROAD) was started on November 1. This was followed with thalidomide and radiation therapy of the right inguinal region was added. On December 16th, she suddenly experienced speech disturbance, nausea and the disturbance of consciousness. Examination of her cerebrospinal fluid showed 368/microl mononuclear cells with 93% plasma cells. The plasma cells disappeared after the 6th intrathecal injection with MTX and prednisolone and the chemotherapy was resumed. One month later, CNS relapse was apparent followed by generalized spread of the tumor mass, and she died on March 17, 2003. PMID- 15553050 TI - [Relapse of diffuse large B cell lymphoma to CD20-negative multiple cutaneous tumors immediately after anti-CD20 monoclonal antibody (rituximab) therapy]. AB - A 60-year-old male was referred to our hospital because of cervical lymphadenopathy and a left hilar abnormal shadow seen on chest X-ray in May 1999. The pathological findings of the cervical lymph nodes revealed that the patient had a malignant lymphoma, of the diffuse large B cell type, at clinical stage IIIB. Immunohistochemistry demonstrated that the lymphoma cells were positive for CD11a, CD19, CD20, CD23, CD25, CD45, IgM, IgD and lambda, but negative for CD5. Although a complete remission was obtained after 8 courses of CHOP therapy, the patient relapsed 32 months later. Two courses of a half dose of CHASE therapy consisting of CPM, ara-C, VP-16 and dexamethasone, followed by rituximab (600 mg/week x4) resulted in a transient re-induction of complete remission. However, multiple cutaneous tumors became apparent just 10 days after the last rituximab therapy. Immunohistochemistry of the cutaneous tumors revealed infiltration of CD20-negative lymphoma cells. A series of chemotherapy including high-dose MTX was ineffective, and the patient died in August 2003. Autopsy findings revealed the systemic intra-capillary infiltration of CD20 negative-lymphoma cells into multiple organs, including the lungs, liver, and kidneys. A CD20 negative-clone selected by rituximab therapy appeared to have expanded in this case. PMID- 15553051 TI - [Myelodysplastic syndrome associated with intestinal tract-type Behcet disease characterized by an esophageal ulcer]. AB - A 55-year-old man with advanced myelodysplastic syndrome was hospitalized prior to undergoing an allogeneic bone marrow transplantation. Immediately before hospitalization, he had suffered from phlegmon in both lower extremities and right forearm as well as genital and oral ulcers. After admission, he developed an esophageal ulcer and was thus diagnosed as having intestinal tract-type Behcet disease. HLA-B51 was not present. Within a month, he died of pulmonary hemorrhage associated with pneumonia, possibly because of a low platelet count, and vasculoendothelial damage related to Behcet disease. This is a rare case of myelodysplastic syndrome that developed Behcet disease with a severe esophageal ulcer. PMID- 15553052 TI - [Pseudomonas sepsis with ecthyma gangrenosum in an acute myeloid leukemia patient]. AB - A 56-year-old woman with acute myeloid leukemia had two rapidly growing necrotizing nodules with ulcer formation on her head after the first course of consolidation therapy. Clinical features corresponding to sepsis (e.g., fever) appeared following the development of the skin lesion. Pseudomonas aeruginosa was isolated from the blood as well as pus of the lesion. Based on these findings, a diagnosis of ecthyma gangrenosum was made. Treatment with ciprofloxacin and gamma globulin dramatically improved the patient's clinical features. Since Pseudomonas sepsis with ecthyma gangrenosum is associated with a high mortality rate, it is important to start immediate treatment with appropriate antibiotics. PMID- 15553055 TI - Promoting evidence-based practice in spinal cord injury/disorders health care. PMID- 15553056 TI - Lessons learned in implementing SCI clinical practice guidelines. AB - While clinical practice guidelines (CPGs) were designed as a tool to improve patient outcomes, decrease practice variation, and optimize resource utilization, providers often encounter significant barriers to integrating these into clinical practice. A study was conducted at six spinal cord injury (SCI) centers in the Department of Veterans Affairs (VA) to improve provider adherence and patient outcomes of two CPGs: Prevention of Thromboembolism in Spinal Cord Injury and Neurogenic Bowel Management in Adults With Spinal Cord Injury. To design effective implementation strategies, focus groups were conducted to identify provider-perceived barriers and facilitators to implementing recommendations for each of the SCI guidelines. Based on this information, four guideline implementation strategies were designed: (a) use of local opinion leaders ("clinical champions"), (b) patient-mediated interventions, (c) standardized documentation template/standing orders, and (d) social marketing/outreach visits. These strategies were implemented at each site. This article identifies "lessons learned" during the process of trying to get these CPGs embedded into clinical practice. PMID- 15553057 TI - Improving respiratory vaccination rates in veterans with spinal cord injury/disorders: lessons learned. AB - The burden of respiratory disease following a spinal cord injury/disorder (SCI/D) is well-described and experienced by patients, families, health care providers, and the health care delivery system. Despite the effectiveness of respiratory vaccinations (influenza and pneumococcal), vaccination rates in this population have been historically low. The Veterans Health Administration (VHA) Spinal Cord Injury Quality Enhancement Research Initiative (QUERI) developed a program to increase vaccination rates for veterans with SCI/D. Patient, provider, and system level strategies were used. The purpose of the study was to improve vaccination rates in this population using multiple strategies. The result was significant improvement in vaccination rates following intervention. This paper describes the lessons learned in implementing best practices for vaccination. In addition, the basic elements of this program, and the similarities with the Chronic Care Model, are described. PMID- 15553058 TI - Understanding a VA preventive care clinical reminder: lessons learned. AB - This article presents "lessons learned" about using a computerized clinical reminder (CCR) for influenza in spinal cord injury (SCI) centers in the Veterans Health Administration (VHA). Although research has shown that use of a CCR increases adherence to preventive care measures, technical and administrative issues affect its use by nurses. CCR is a term for a reminder in a patient's electronic medical record to initiate a preventive care activity. Qualitative data about using the CCR for influenza were collected from staff using semi structured interviews, e-mail questionnaires, and telephone calls. Once issues were identified, follow-up activities included discussions with local and national experts and problem-solving activities. The investigators identified two national issues about the CCR for influenza: the taxonomy to identify patients with SCI, and the "due date" for influenza vaccine. Local issues included authorization for nurses to use the CCR, ease of use of the CCR for inpatients and outpatients, connectivity of the medication tracking program for inpatients with the electronic medical record, configuration of the CCR in the electronic medical record, training, and technical support. PMID- 15553059 TI - Staff influenza vaccination: lessons learned. AB - Respiratory complications are the top cause of morbidity and mortality among persons with spinal cord injury/disorders (SCI/D). One method to protect patients with SCI/D from complications is to reduce exposure to influenza through vaccination of health care workers (HCWs). This study examined the extent to which HCWs promoted safe, quality care for patients with SCI/D as related to their beliefs about, and receipt/non-receipt, of influenza vaccination. Surveys were distributed to HCWs at 23 SCI/D centers within the Veterans Health Administration (VHA). Vaccination rates, reasons for receipt/non-receipt, and possible associations were examined. The response rate was 53% (n = 820). The SCI/D staff influenza vaccination rate was 50%, with the lowest rates among nurses. The most common reason for vaccination was self-protection. The most common reason for not being vaccinated was concern about side effects. Of providers who recommended the vaccine to patients, significantly more (60%) received the vaccine; p = .000. PMID- 15553060 TI - In search of the "superior" cervical orthosis: Philadelphia Cervical Orthosis versus Aspen Cervical Orthosis. PMID- 15553061 TI - Qualitative research: where do nurses begin? PMID- 15553062 TI - Leading through partnering: from bedside to community. AB - Partnering as a means of leading requires a particular focus and has particular characteristics. It is unrealistic to think that every person that participates in a partnership would have honed the skills to provide guidance, strength, and support for the process. It is not likely that every partner understands the collaborative process well enough to engage all partners with tact, openness, fairness, and critical, but respectful, reflection. The characteristics depicted in the Leading Through Partnering dome reflect those leaders who have integrated partnering into a coherent framework of action. Stern (2003), in describing her grounded theory research on "attentive partnering" among colleagues, determined that conditions for partnering seem to require the presence of "determined, persuasive leaders who foster growth-enhancing collegial relationships" (pg. 271). The concept of partnering continues to take hold in many forms. Leading Through Partnering as a variant form, whether occurring on a small scale at the bedside or a large scale in the community, is likely to be more than just a passing trend. PMID- 15553063 TI - Educating children, adolescents, and their families following spinal cord injury. PMID- 15553064 TI - The nursing shortage and its impact on retention and recruitment. AB - According to Nevidjon and Erickson (2001), 1.8 million nurses work primarily in hospital settings where the shortage is greatest, but the impact of the nursing shortage is also having major effects in all health care settings. This review looked at a few short- and long-term solutions that will need to be implemented to resolve the problem. Professional organizations, such as JCAHO and ANA, are joining forces to keep nursing shortage issues in the forefront. The government is keenly aware of the problem and is providing funds to address the nursing shortage. Even so, by now, it should be glaringly clear to the reader that there are no easy solutions. It will take the combined efforts of everyone to bring about a positive change for the future of the nursing profession. PMID- 15553065 TI - [Viral hepatitis type C: new facts and hypotheses]. AB - It has been generally accepted that cell-mediated immunity is critical to both the resolution of acute HCV infection and the pathogenesis of chronic hepatitis. However, they are a lot questions still waiting for elucidation. This article is a summary of recent facts and hypotheses formulated in last two years in the field of investigations on viral hepatitis C. PMID- 15553066 TI - [Treatment strategies of acute and chronic hepatitis C]. AB - There is a overview of actual regimens proposed for treatment of acute and chronic hepatitis C. The new formulations of interferon alpha obtained by pegylation--PEG-IFN-s have resulted in improved efficacy as monotherapy and in combination with ribavirin. The results of clinical trials and therapeutical recommendations have been presented. PMID- 15553067 TI - [Cholestatic liver disease in children]. AB - Cholestatic liver disease constitutes a large part of chronic liver diseases during infancy. It is caused either by extrahepatic disorders (obstruction) or by intrahepatic cholestasis (functional). The differential diagnosis should be done as early as possible because the delayed surgical therapy in extrahepatic cholestasis has a very bad prognosis. Intrahepatic cholestasis may be caused by a broad spectrum of different disorders such as congenital infection, endocrine, chromosomal abnormalities or inborn errors of metabolism. Familial clustering is typical for the Byler's disease, Alagille's syndrome, PFIC, errors of bile acid synthesis or alpha-1-ATD. The established diagnosis allows to start etiological treatment: dietary--in metabolic diseases, antibiotics--in bacterial infections, antiviral--in viral infections etc. To lower bile acids level pharmacological treatment (UDCA) or surgical procedures (hepatoportoenterostomy, partial biliary diversion) should be performed. In progressive cholestasis the only effective therapy is liver transplantation. PMID- 15553068 TI - [Two cases of Wilson disease diagnosed as autoimmune hepatitis]. AB - Authors present two cases of Wilson disease in which the final diagnosis was very difficult and needed the differentiation with autoimmune hepatitis. Observation of those two patients leads to the conclusion that paediatricians have to deepen they knowledge of autoimmune hepatitis and Wilson disease which are seen more often in children now-a-days. It will help to quicker the diagnosis and treatment which is most essential for better prognosis in those patients. PMID- 15553069 TI - [Endoscopy in the diagnosis of chronic hepatitis C--diagnosis and therapy depending on the phase of diseases progression and liver regeneration]. AB - The GI endoscopy can be divided into upper GI tract endoscopy (esophago-, gastro , entero-, fistulo- and cholangioscopy) and lower GI tract endoscopy (recto-, sigmoido-, colonoscopy) from practical point of view and the characteristic of used equipment. A lot of therapeutic methods for GI tract is associated with each of these procedures. GI tract endoscopy doesn't play significant part in diagnosis of acute and chronic C hepatitis. Significance of endoscopy procedures decidedly increases in the case of progressive liver fibrosis and liver cirrhosis associated with HCV infection., where changes in GI tract are observed to 87% patients. These changes can be divided into: 1) not associated with portal hypertension, 2) these ones caused by portal hypertension. The most observed changes not associated with portal hypertension involve: reflux esophagitis, esophageal candidiasis; different variants of gastritis, gastric and duodenal ulcer. To the changes connected with portal hypertension, which are possible for endoscopy assessment, belongs esophageal and gastric varices, portal gastro-, entero-, colopathy, and gastric antral vascular ectasiae (GAVE). However to-day endoscopy has got not only diagnostic significance but also enables: estimation of pharmacotherapy efficiency, the primary and secondary prophylaxis of bleedings from GI varices as well as therapy of GI bleeding in this group of patients. PMID- 15553070 TI - [Ribavirin--pharmacological features, antiviral effects against hepatitis C virus (HCV) and other viruses and side effects during treatment]. AB - Ribavirin is a broad spectrum antiviral agent first synthesised in 1972. It shows varying degrees of clinical efficacy in a variety of human viral infections including viral hepatitis, respiratory tract infections and haemorrhagic fevers. This review focuses on the complicated pharmacokinetics of ribavirin as well as on its direct (antiviral) and indirect (immunomodulatory) modes of action with particular emphasis on its antiviral activity against HCV. Side effects and drug interactions have also been briefly described. PMID- 15553071 TI - [Chronic hepatitis C in HIV-infected patients receiving highly active antiretroviral therapy (HAART)]. AB - Introduction of HAART in the treatment of HIV-infection revealed, that clinical consequences of HIV-infection in these patients occur earlier than in HIV negative ones. It may be a reason of unsuccessfully antiretroviral therapy. Another problem is hepatotoxicity of antiretroviral therapy, especially in HCV infected patients. In the follow, treatment of chronic hepatitis C seems to be justified. PMID- 15553072 TI - [HGV, TTV and SEN viruses--are these organisms pathogenic?]. AB - After the discovery of hepatitis A, B, C, D and E viruses, several lines of evidence suggested that an additional hepatitis agent (s) might exist. Molecular approaches led sequentially to the discoveries of GBV-C/HGV, TTV and more recently SENV. At issue is whether these agents cause non - A non - E hepatitis and/or some extrahepatic diseases. These problems are discussed in present paper, considering the newest data from literature. PMID- 15553073 TI - [Acute hepatitis C]. AB - OBJECTIVE: The aim of the study was both to evaluate the natural outcome of acute hepatitis C (AHC), in this factors influencing chronicity of HCV, and to assess results of antiviral therapy in acute phase of disease. PATIENTS AND METHODS: Seventy-seven pts with diagnosis of AHC (all HCVRNA-positive, in this 44 anti-HCV positive at entry) were seen. Sixty-four of them (F/M: 33/31; mean age: 45.4y +/- 16.0) were non-treated (NT pts) with IFN during 6-months follow-up and thirteen patients (F/M: 5/8; mean age: 36.6ys +/- 11.9) were treated (T pts) with IFN. Antiviral therapy was started within 2 to 18 (median 10) weeks after the onset of AHC including seven pts with IFN alpha-2b at 5MU thrice weekly (TIW) and one pt with IFN-alpha-2a at 6MU TIW for 24 weeks, three pts with combined (Rebetron) therapy according to HCV genotype (1F/3a for 24 weeks and 2M/1b for 48 weeks) and two pts with PEG-Intron for one month followed by Intron at 3MU TIW for 20 weeks. RESULTS: 64.1% of NT pts developed chronic hepatitis. In multivariate logistic regression analysis age > 40y, nonicteric course and medical procedures as route of infection were independently and significantly associated with chronicity. Ten of the thirteen (77%) treated pts presented sustained response (SR) at the end of follow-up. Only one seronegative pt no responded to the therapy. The others 12 pts (all seropositive) presented rapid normalisation of ALT levels. However, 3 of them developed breakthrough between 12 and 20 week of therapy and ribavirin was added with beneficial effect. At the end of treatment point, all seropositive patients showed biochemical and virological response but two (with breakthrough history) of them relapsed. CONCLUSIONS: Seropositive, nonicteric patients with acute hepatitis C should be treated as soon as possible and pegylated-IFN for 24 weeks seems to be currently the best choice. Combination therapy with PEG and ribavirin could be proposed to the patients who failed to monotherapy. The beneficial effect of AHC therapy is indicative for necessity of creating conditions for diagnosed AHC more frequent than hitherto. PMID- 15553074 TI - Who will educate the next generation of nurses? A looming faculty shortage. AB - Nursing is a profession that has many career choices. This article highlighted some of the major issues that will affect practicing hospital nurses and nurse educators. The nursing shortage is no longer "impending"-it is here and there are no signs that there will be a reprieve for staffing at the bedside. The major point of concern made in this article is that we must prompt our best and brightest to consider a career in nurse education. We are leaving too many qualified applicants to nursing school behind due to a faculty shortage in U.S. schools and colleges of nursing. Every nurse is called upon to "think outside the box" and employ strategies of collaboration between the practice arena [whether hospital, long term care, or hospice] and the academic setting to manage the shortage of nurses and nurse faculty. Using existing resources in a different way may assist in educating our future nurses. PMID- 15553075 TI - Atypical spinal cord injury: spinal dural arteriovenous fistula. AB - Spinal dural arteriovenous fistulas (SDAVF) are vascular anomalies composed of intertwining arteries and veins with direct arteriovenous (AV) communication. It is presumed that the fistula is an acquired abnormality that produces an arterialization and increase in venous blood flow leading to venous hypertension, venous congestion with eventual hypo-perfusion, or ischemia of the spinal cord. Symptoms include progressive sensory and motor changes that commonly take place over a 2- to 3-year time span. Exacerbation of symptoms is frequently the reason for additional diagnostic work-up. Treatment is aimed at elimination of the communication between arteries and veins. Treatment options include surgical resection of the piece of dura containing the AV fistula, transvascular embolization, or a combined procedure. Outcome is a function of the extent of cord injury and full recovery is rare. Rehabilitation is crucial in maximizing functional outcomes. It is essential for the nurse caring for the patient with a SDAVF to understand the pathological changes related to the fistula, correlate clinical findings, identify diagnostic tools for evaluation, and differentiate treatment options. An understanding of the disorder will provide the groundwork for the nurse to formulate a plan of care identifying patient needs from assessment parameters, rehabilitation needs, and patient education. PMID- 15553076 TI - A model for assessing learning readiness for self-direction of care in individuals with spinal cord injuries: a qualitative study. AB - Methods are needed for assessing and facilitating learning readiness for self direction of care of individuals with spinal cord injury (SCI). With declining length of stay, self-direction of care is a major learning task for patients with permanent disability. A retrospective study of secondary data gathered from 30 nursing records was used to develop the Rehabilitation Learning Readiness Model for Spinal Cord Injury. The framework evolved from Orem's (1995) self-care deficit nursing theory and two self-directed adult learning models (Grow, 1991; Pratt, 1988). Using qualitative methods of analysis, five stages of learning readiness behavior (Dependent, Involvement, Engagement, Self-Initiation, and Self Direction) and five corresponding nursing roles (Authority, Guide, Motivator, Mentor, and Consultant), emerged. Expert nurses corroborated the clinical significance of the model. The model is proposed as a conceptual paradigm for use by nurses to assess learning readiness challenges experienced by patients with SCI and to improve teaching roles and practices. PMID- 15553077 TI - Understanding care-related abuse and neglect in the lives of women with SCI. AB - Abuse and neglect of women with spinal cord injury (SCI) is a serious health issue. To begin to understand the nature of this problem an interpretive phenomenological study of abuse of women with SCI was conducted. A sample of 13 women who had experienced abuse post-injury participated in a total of 24 in depth interviews. Analytic strategies employed included thematic analysis, exemplars, biographical comparisons, and paradigm cases. The analysis yielded four major categories including (a) abuse and neglect-definitions and types, (b) vulnerability factors, (c) behavioral responses to abuse, and (d) health effects. This research report reviews findings from the first category, abuse definitions and types. Findings indicate that care-related forms of abuse and neglect are particularly common in the lives of women with SCI. PMID- 15553078 TI - The adaptation of children to spinal cord injury of a family member: the individual's perspective. AB - The importance of family involvement in spinal cord injury (SCI) rehabilitation is well-documented, but little is known about the needs of children in these families. This study surveyed persons with SCI at a national rehabilitation center to establish the number who had close relatives who were children, gain their views on facilities available for visiting children, and determine the impact of SCI on children. Thirty-two inpatients provided details of any close relatives who were children, and 17 participants supplied further information in a semi-structured interview. Twenty-six participants (81%) had close relatives under age 18, for a total of 88 children. Most children had visited participants at the centre, but levels of satisfaction with facilities were low. Participants also commented on children's understanding of SCI and coping strategies. The results suggest that rehabilitation teams could consider children's needs more fully, provide appropriate physical facilities, and facilitate understanding and emotional adjustment of children. PMID- 15553079 TI - Reproductive health for women with spinal cord injury: pregnancy and delivery. PMID- 15553080 TI - Crossing the limits: interstate licensure. PMID- 15553081 TI - De-mystifying the expert witness testimony experience. PMID- 15553082 TI - Breaking through the glass ceiling: women in executive leadership positions--part II. PMID- 15553083 TI - Strategies for coping with the non-compliant adolescent spinal cord injury patient--part II. PMID- 15553084 TI - Gene rearrangements and evolution of tRNA pseudogenes in the mitochondrial genome of the parrotfish (Teleostei: Perciformes: Scaridae). AB - Genomic size of animal mitochondrial DNA is usually minimized over time. Thus, when regional duplications occur, they are followed by a rapid elimination of redundant material. In contrast to this general view, we report here long sustained tRNA pseudogenes in the mitochondrial genome (mitogenome) of teleost fishes of the family Scaridae (parrotfishes). During the course of a molecular phylogenetic study of the suborder Labroidei, we determined the complete nucleotide sequence of the mitogenome for a parrotfish, Chlorurus sordidus, and found a gene rearrangement accompanied by a tRNA pseudogene. In the typical gene order of vertebrates, a tRNA-gene cluster between ND1 and ND2 genes includes tRNA(Ile) (I), tRNA(Gln) (Q), and tRNA(Met) (M) genes in this order (IQM). However, in the mitogenome of the parrotfish, the tRNA(Met) gene was inserted between the tRNA(Ile) and the tRNA(Gln) genes, and the tRNA(Gln) gene was followed by a putative tRNA(Met) pseudogene (psiM). Such a tRNA gene rearrangement including a pseudogene (IMQpsiM) was found in all of the 10 examined species, representing 7 of the 10 currently recognized scarid genera. All sister groups examined (20 species of Labridae and a single species of Odacidae) had the typical gene order of vertebrate mitogenomes. Phylogenetic analysis of the tRNA(Met) genes and the resulting pseudogenes demonstrated that the ancestral tRNA(Met) gene was duplicated in a common ancestor of the parrotfish. Based on the fossil record, these results indicate that the pseudogenes have survived at least 14 million years. Most of the vertebrate mitochondrial gene rearrangements involving the IQM region have held the tRNA(Met) gene just upstream of the ND2 gene, and even in a few exceptional cases, including the present ones, the tRNA pseudogenes have been found in that position. In addition, most of these tRNA(Met) pseudogenes maintained clover-leaf secondary structures, with the remainder sustaining the clover-leaf structure in the "top half (TpsiC and acceptor arms). Considering their potential secondary structures (holding "top halves" of the clover-leaf structures), locations within mitogenomes (flanking the 5' ends of the ND2 genes) and stabilities over time (survived at least 14 Myr), it is likely that the tRNA pseudogenes retain function as punctuation marks for mitochondrial ND2 mRNA processing. PMID- 15553085 TI - Teleost fish genomes contain a diverse array of L1 retrotransposon lineages that exhibit a low copy number and high rate of turnover. AB - Retrotransposable elements exhibit a wide range of variation in population dynamics, abundance, and lineage diversity among host genomes across taxa. This range of diversity is illustrated by a single well-defined constituent monophyletic clade of L1 non-LTR retrotransposons that is shared between mammalian and teleost fish genomes. Despite the clear phylogenetic relationships that exist between mammalian and teleost L1 sequences, these elements exhibit markedly different dynamics within their respective taxa. While mammalian genomes typically contain a single, abundant lineage of L1 elements that traces millions of years of evolution, the zebraflsh genome was recently shown to exhibit a high diversity of ancient lineages coexisting at a very low copy number and apparently exhibiting a high rate of turnover. In the present study, a combination of degenerate PCR, lineage-specific PCR, and genomic Southern blot analysis is utilized to demonstrate high L1 lineage diversity, low copy number, and a high proportion of polymorphic inserts in the genomes of the killifish species, Fundulus heteroclitus. Additional species surveyed by degenerate PCR include Cyprinodon variegatus, Rivulus marmoratus, and Menidia beryllina. These results further support the generality of the differences that exist in host-element dynamics between teleost fish and mammalian genomes with regard to L1 retrotransposons. PMID- 15553086 TI - Measuring the coding potential of genomic sequences through a combination of triplet occurrence patterns and RNY preference. AB - The distribution of n-tuplet frequencies is shown to strongly correlate with functionality when examining a genomic sequence in a reading-frame specific manner. The approach described herein applies a coarse-graining procedure, which is able to reveal aspects of triplet usage that are related to protein coding, while at the same time remaining species independent, based on a simple summation of suitable triplet occurrences measures. These quantities are ratios of simple frequencies to suitable mononucleotide-frequency products promoting the incidence of the RNY motif, preferred in the most widely used codons. A significant distinction of coding and noncoding sequences is achieved. PMID- 15553087 TI - Interspecific chromosome-wide transcription profiles reveal the existence of mammalian-specific and species-specific chromosome domains. AB - A long-range exploration of expression levels through wide chromosome territories was carried out in three species (pig, cattle, and chicken) by aligning EST counts against the human genome. This strategy made it possible to produce expression profiles that were very similar between pig and cattle and that were significantly correlated with chicken levels of expression. In parallel with these alignments, we developed a statistical approach enabling us to screen genomic regions for both underexpression and overexpression at the chromosome level within a given species, as well as interspecifically. The observed correlations are indicative of the existence of interspecifically conserved domains of gene expression, not only for housekeeping genes (which are highly expressed), but also for regions where genes are significantly underexpressed. Furthermore, our strategy made it possible to point out regions that are differentially regulated between species. These expression data were crossed with available comparative mapping information for pigs and cattle, suggesting that coregulated regions are syntenic in various mammals. PMID- 15553088 TI - Comparative genetics of functional trinucleotide tandem repeats in humans and apes. AB - Several human neurodegenerative disorders are caused by the expansion of polymorphic trinucleotide repeat regions. Many of these loci are functional short tandem repeats (STRs) located in brain-expressed genes, and their study is thus relevant from both a medical and an evolutionary point of view. The aims of our study are to infer the comparative pattern of variation and evolution of this set of loci in order to show species-specific features in this group of STRs and on their potential for expansion (therefore, an insight into evolutionary medicine) and to unravel whether any human-specific feature may be identified in brain expressed genes involved in human disease. We analyzed the variability of the normal range of seven expanding STR CAG/CTG loci (SCA1, SCA2, SCA3-MJD, SCA6, SCA8, SCA12, and DRPLA) and two nonexpanding polymorphic CAG loci (KCNN3 and NCOA3) in humans, chimpanzees, gorillas, and orangutans. The study showed a general conservation of the repetitive tract and of the polymorphism in the four species and high heterogeneity among loci distributions. Humans present slightly larger alleles than the rest of species but a more relevant difference appears in variability levels: Humans are the species with the largest variance, although only for the expanding loci, suggesting a relationship between variability levels and expansion potential. The sequence analysis shows high levels of sequence conservation among species, a lack of correspondence between interruption patterns and variability levels, and signs of conservative selective pressure for some of the STR loci. Only two loci (SCA1 and SCA8) show a human specific distribution, with larger alleles than the rest of species. This could account, at the same time, for a human-specific trait and a predisposition to disease through expansion. PMID- 15553089 TI - Gene network polymorphism is the raw material of natural selection: the selfish gene network hypothesis. AB - Population genetics, the mathematical theory of modern evolutionary biology, defines evolution as the alteration of the frequency of distinct gene variants (alleles) differing in fitness over the time. The major problem with this view is that in gene and protein sequences we can find little evidence concerning the molecular basis of phenotypic variance, especially those that would confer adaptive benefit to the bearers. Some novel data, however, suggest that a large amount of genetic variation exists in the regulatory region of genes within populations. In addition, comparison of homologous DNA sequences of various species shows that evolution appears to depend more strongly on gene expression than on the genes themselves. Furthermore, it has been demonstrated in several systems that genes form functional networks, whose products exhibit interrelated expression profiles. Finally, it has been found that regulatory circuits of development behave as evolutionary units. These data demonstrate that our view of evolution calls for a new synthesis. In this article I propose a novel concept, termed the selfish gene network hypothesis, which is based on an overall consideration of the above findings. The major statements of this hypothesis are as follows. (1) Instead of individual genes, gene networks (GNs) are responsible for the determination of traits and behaviors. (2) The primary source of microevolution is the intraspecific polymorphism in GNs and not the allelic variation in either the coding or the regulatory sequences of individual genes. (3) GN polymorphism is generated by the variation in the regulatory regions of the component genes and not by the variance in their coding sequences. (4) Evolution proceeds through continuous restructuring of the composition of GNs rather than fixing of specific alleles or GN variants. PMID- 15553090 TI - Functional constraints of 6-phosphogluconate dehydrogenase (6-PGD) based on sequence and structural information. AB - The pentose phosphate cycle is considered as a major source of NADPH and pentose needed for nucleic acid biosynthesis. 6-Phosphogluconate dehydrogenase (6PGD), an enzyme participating in this cycle, catalyzes the oxidative decarboxylation of 6PGD to ribulose 5-phosphate with the subsequent release of CO2 and the reduction of NADP. We have determined the amino acid sequence of 6PGD of Bactrocera oleae and constructed a three-dimensional model based on the homologous known sheep structure. In a comparative study of 6PGD sequences from numerous species, all the conserved and variable regions of the enzyme were analyzed and the regions of functional importance were localized, in an attempt promoted also by the direct involvement of the enzyme in various human diseases. Thus, analysis of amino acid variability of 37 6PGD sequences revealed that all regions important for the catalytic activity, such as those forming the substrate and coenzyme binding sites, are highly conserved in all species examined. Moreover, several amino acid residues responsible for substrate and coenzyme specificity were also found to be identical in all species examined. The higher percentage of protein divergence is observed at two regions that accumulate mutations, located at the distant parts of the two domains of the enzyme with respect to their interface. These peripheral regions of non-functional importance are highly variable and are predicted as antigenic, thus reflecting possible regions for antibody recognition. Furthermore, locating the differences between diptera 6PGD sequences on the three-dimensional model suggests probable positions of different amino acid residues appearing at B. oleae fast, intermediate, and slow allozymic variants. PMID- 15553091 TI - Position-associated GC asymmetry of gene duplicates. AB - It is well known that repositioning of a gene often exerts a strong impact on its own expression and whole development. Here we report the results of genome-wide analyses suggesting that repositioning may also radically change the evolutionary fate of gene duplicates. As an indicator of these changes, we used the GC content of gene pairs which originated by duplication. This indicator turned out to be duplicate-asymmetric, which means that genes in a pair differ significantly in GC content despite their apparent origin from a common ancestor. Such an asymmetry necessarily implies that after duplication two originally identical genes mutated in opposite directions-toward GC-rich and GC-poor content, respectively. In mammalian genomes, this trend is definitely associated with presumably methylated hypermutable CpG sites, and in a typical GC-asymmetric gene pair, its two member genes are embedded in GC-contrasting isochores. However, we unexpectedly found similar significant GC asymmetry in fish, fly, worm, and yeast. This means that neither methylation alone nor methylation in combination with isochores can be counted as a primary cause of the GC asymmetry; rather they represent specific realizations of some universal principle of genome evolution. Remarkably, genes from pairs with the greatest GC asymmetry tend to be on different chromosomes, suggesting that the mutational difference between gene duplicates is associated with translocation of a new gene to a different place in the genome, whereas GC symmetric pairs demonstrate the opposite tendency. A recently emerged extra gene copy is usually on the same chromosome as is its parent but quickly, by 0.05 substitution per synonymous site, either has perished or occupies a different chromosome. During this earliest posttranslocation period, the ratio of nonsynonymous/synonymous base substitutions is unusually high, suggesting a rapid adaptive evolution of novel functions. In a general context of evolution by gene duplication, our interpretation of this position-dependent GC asymmetry between duplicated genes is that evolution of redundant genes toward a new function has often been associated with their very early, postduplication repositioning in the genome, with a concomitant abrupt change in epigenetic control of tissue/stage specific expression and an increase in the mutation rate. Of eight eukaryotic genomes studied, the most distinguished in this respect is the human genome. PMID- 15553092 TI - Modulation of base-specific mutation and recombination rates enables functional adaptation within the context of the genetic code. AB - The persistence of life requires populations to adapt at a rate commensurate with the dynamics of their environment. Successful populations that inhabit highly variable environments have evolved mechanisms to increase the likelihood of successful adaptation. We introduce a 64 x 64 matrix to quantify base-specific mutation potential, analyzing four different replicative systems, error-prone PCR, mouse antibodies, a nematode, and Drosophila. Mutational tendencies are correlated with the structural evolution of proteins. In systems under strong selective pressure, mutational biases are shown to favor the adaptive search of space, either by base mutation or by recombination. Such adaptability is discussed within the context of the genetic code at the levels of replication and codon usage. PMID- 15553093 TI - Selection on codon usage for error minimization at the protein level. AB - Given the structure of the genetic code, synonymous codons differ in their capacity to minimize the effects of errors due to mutation or mistranslation. I suggest that this may lead, in protein-coding genes, to a preference for codons that minimize the impact of errors at the protein level. I develop a theoretical measure of error minimization for each codon, based on amino acid similarity. This measure is used to calculate the degree of error minimization for 82 genes of Drosophila melanogaster and 432 rodent genes and to study its relationship with CG content, the degree of codon usage bias, and the rate of nucleotide substitution. I show that (i) Drosophila and rodent genes tend to prefer codons that minimize errors; (ii) this cannot be merely the effect of mutation bias; (iii) the degree of error minimization is correlated with the degree of codon usage bias; (iv) the amino acids that contribute more to codon usage bias are the ones for which synonymous codons differ more in the capacity to minimize errors; and (v) the degree of error minimization is correlated with the rate of nonsynonymous substitution. These results suggest that natural selection for error minimization at the protein level plays a role in the evolution of coding sequences in Drosophila and rodents. PMID- 15553095 TI - [Bio-artificial organs]. PMID- 15553094 TI - Molecular adaptation in plant hemoglobin, a duplicated gene involved in plant bacteria symbiosis. AB - The evolutionary history of the hemoglobin gene family in angiosperms is unusual in that it involves two mechanisms known for potentially generating molecular adaptation: gene duplication and among-species interaction. In plants able to achieve symbiosis with nitrogen-fixing bacteria, class 2 hemoglobin is expressed at high concentrations in nodules and appears to be a key factor for the achievement and regulation of the symbiotic exchange. In this study, we make use of codon models of DNA sequence evolution with the goal of determining the nature of the selective forces which have driven the evolution of this gene. Our results suggest that adaptive evolution occurred during the period of time following the duplication event (functional divergence) and that a change in the selective pressures arose in class 2 hemoglobin in relation to the acquisition of a symbiotic function. PMID- 15553096 TI - [Biomaterials]. AB - Biomaterials are essential components in the development of artificial organs. Synthetic polymers are widely used for a number of biomedical applications: a) as medical supplies, b) devices to support or replace malfunctioning body parts or c) systems with a local therapeutic function. The design of biomaterials has developped over the past three decades from devices with a mechanical function, over systems with a dedicated function (degradable, drug releasing, surface modified) to devices which are tailor made to interact in a smart way with the biological environment. At present polymers are attractive materials to construct scaffolds for tissue regeneration and regenerative medicine. The material design is adapted to the requirements for cell ingrowth, cell migration, angiogenesis etc. This is mainly controlled by the surface topography and chemistry that play a key role in the interfacial material-cell communication. This remarkable development is the direct result of a multi-disciplinary approach, combining material science with engineering, cell biology, molecular biology, medicine and regulatory guidelines. PMID- 15553097 TI - [Bio-artificial organs: cardiac applications]. AB - The most important bio-prosthetic organs in cardiovascular medicine are artificial heart valve prostheses and blood pumps. Cardiac valve prostheses can be divided in 'mechanical' and 'biological' valves. Mechanical prostheses are entirely made of artificial materials and require a meticulous anticoagulation therapy. The biological heart valves or heterografts (the allo- and autografts are not considered in this issue) are made of fixed or crosslinked biological tissue and therefore anticoagulation of the patient is not necessary. Actual mechanical heart valves consist of a titanium ring in which one or more leaflets made of pyrolite carbon assure the opening and closing mechanism. Biological heart valves are made of fixed porcine aortic valves or of fixed bovine pericardium. Both tissues can be mounted on a metal frame (so-called 'stented valves') or they can lack this structure ('stentless valves'). The problem with these biological valves is the durability: during the years they start degenerating or calcifying. To prevent this, recent biological heart valves are treated with an anti-mineralisation procedure. Since recent years intense research is ongoing to develop a living heart valve by 'tissue engineering'. When the entire heart fails, and pharmacological treatment remains inadequate or a heart transplantation is not immediately possible, artificial blood pumps are implanted. In general, two categories of blood pumps can be distinguished: displacement and rotary blood pumps. Displacement pumps move a certain amount of blood by the movement of a flexible membrane. This movement is pneumatically or electrically driven and requires an extensive installation: artificial ventricles, tubing, motors, pneumatic systems and driving consoles. Examples in clinical use are: Novacor, Heart Mate, Thoratec, Medos/HIA. Recent developments focus on miniaturisation and endovascular implantation. The rotary blood pumps are well suited for these purposes. They can be either axial, diagonal or radial pumps. A promising new develoment is the Impella pump: this is an axial flow pump having a diameter of 4 mm, which is implanted, inclusive the electrical motor, in the failing heart and delivers an output of 2 to 10 liter/minute. Clinical testing of this device is ongoing. PMID- 15553098 TI - Cochlear implantation: a bionic system to restore hearing in severely hearing impaired patients. AB - CI replaces and functionally bypasses the inner haircells of the cochlea. Candidate selection is not merely based on non-aided auditory thresholds, but aided speech recognition tests in quite and in noise after a rehabilitation period may be decisive, besides a comprehensive selection process including general health, imaging, learning and cognitive skills, supportive aspect of the environment. The vast majority of severely or profoundly hearing impaired persons can benefit from CI. CI restores hearing of adults to a level allowing normal conversation, using the telephone and even understanding in a limited amount of background noise. Congenitally severely or profoundly hearing impaired children implanted before the age of 2 years, will outperform patients operated at a later age, allowing to follow mainstream education. CI is not just a technique, but the driving force behind an elaborate care-programme for the severely hearing impaired patient. PMID- 15553099 TI - [Bio-artificial implants in orthopedics]. AB - Aging of the population and thus of loadbearing cartilage produces a number of degenerative symptoms. At the knee, considerable macroscopic asymmetry of the joint underlies these degenerative changes, were it not for two menisci that somewhat neutralize this discongruency. Microscopically, cartilage is a structure within the knee joint that allows for low friction and congruent movement. Repetitive microtrauma rather than macrotrauma can affect its integrity. Both microscopically and macroscopically reconstruction of a cartilage-like structure is achieved through implantation of cultured cells or auto- and allografts either by minimally invasive or classical surgery, which theoretically could have a beneficial effect on the natural course of degeneration. PMID- 15553100 TI - [Intracellular calcium-releasing channels as cellular targets for immunophilins: a molecular, functional and structural analysis]. AB - In this study, the FKBP12-binding properties of IP3Rs and RyRs were compared. Although the primary sequence of IP3Rs en RyRs contained a putative FKBP12 binding site, the functional, molecular and structural properties of these sites appeared to be completely different. For RyRs, FKBPs appear to function as associated proteins that are important for the functional regulation of the channel, thereby stabilizing the RyR complex. For IP3Rs, FKBPs might be involved in the de novo protein synthesis of the IP3Rs and the folding of the peptide chain to a functional IP3R protein, thereby functioning as helper enzymes. Hence, it is very unlikely that they function as associated regulatory proteins of the IP3R. In addition, we provided evidence that FKBP 12 is also an important regulating protein of the Ca(2+)-flux properties of the RyR3. FKBP12 clearly modulated both RyR3-mediated global and local Ca(2+)-responses. PMID- 15553101 TI - [Recent developments in gene therapy]. AB - Gene therapy is defined as the introduction of genetic material in a patient's cells with resulting therapeutic benefit. It is a promising new biomedical discipline that could potentially lead to new treatments for hereditary diseases, cardiovascular and neurologic disorders, cancer, diabetes and even infectious diseases. The introduction of genetic material into somatic cells requires gene delivery vectors. Since viruses have developed efficient means to introduce their own genetic material into cells they can be readily adapted as viral vectors for gene therapy. Preclinical studies in animal models have shown that therapeutic effects can be achieved after gene therapy for genetic, acquired and complex disorders. Furthermore, therapeutic effects have been obtained in several phase I/II gene therapy clinical trials for hemophilia, severe combined immune deficiency (SCID) and cancer. Gene transfer technology has improved significantly over the past few years and has led to the development of vectors which have fewer side-effects without compromising their efficacy, at least partly due the development of cell-type specific targetable vectors. Nevertheless, the success of gene therapy is still very much depending upon the continuous development of improved vector technologies which would hopefully and ultimately cure diseases which are refractory to current treatment paradigms. PMID- 15553102 TI - Effect of zolpidem on brain injury and diaschisis as detected by 99mTc HMPAO brain SPECT in humans. AB - The study investigates the effect of zolpidem (CAS 82626-48-0) on brain injuries and cerebellar diaschisis. Four patients with varied brain injuries, three of them with cerebellar diaschisis, were imaged by 99mTc HMPAO Brain SPECT before and after application of zolpidem. The baseline SPECT before zolpidem showed poor tracer uptake in brain injury areas and cerebellar diaschisis. After zolpidem, cerebral perfusion through brain injury areas improved substantially in three patients and the cerebellar diaschisis was reversed. Observations point to a GABA based phenomenon that occurs in brain injury and diaschisis that is reversible by zolpidem. PMID- 15553103 TI - Alpha-dihydroergocryptine in the long-term therapy of Parkinson's disease. AB - The trial was designed as an open-label, post-authorisation safety study, aimed to complete the available information on adverse events and drug reactions to alpha-dihydroergocryptine (CAS 14271-05-7, alpha-DHEC). The study included 294 patients with idiopathic Parkinson's disease who received levodopa (CAS 59-92-7, L-DOPA) and started taking alpha-DHEC (Cripar). Adverse events were analysed descriptively, Parkinson's disease symptoms were documented using a questionnaire applied by the physicians. Patients were evaluated at study start and three and six months later, respectively. In 31 patients, 32 adverse events were observed, gastrointestinal and nervous system disorders being the most frequent. Dyskinesias, psychoses/hallucinations, sleep disturbances, and cardiovascular disorders were uncommon (< or = 1%). in total, 21 adverse events were classified as adverse drug reactions. In nearly 80 % of the cases, Parkinson symptoms had improved or completely vanished. Symptoms were unchanged in 16.7 % of patients and had worsened in 3.1%. The results confirm that the use of alpha-DHEC in combination therapy with levodopa in patients with Parkinson's disease is a well tolerated and efficacious treatment option. PMID- 15553104 TI - A rapid, sensitive and validated method for the determination of ondansetron in human plasma by reversed-phase high-pressure liquid chromatography. AB - A simple and sensitive method for the determination of ondansetron (CAS 116002-70 1) in human plasma was developed using high-pressure liquid chromatography (HPLC). The procedure involves extraction of human plasma with tertiary butyl methyl ether containing 2 mol/l sodium hydroxide, followed by reversed-phase HPLC using a LiChrospher 100 RP-18e 5 microm column and UV detection at 305 nm. The retention times of ondansetron and internal standard (propranolol hydrochloride, CAS 318-98-9) were 9.38 and 13.40 min, respectively. The calibration curves were linear over the range of 10 ng/ml (lower limit of quantitation, LOQ) and 380 ng/ml for ondansetron. The intra- and inter-assay coefficients of variation for all the criteria of validation were less than 15% over the linearity range. Ondansetron was stable upon storage in human plasma. The sensitivity and precision of the method were within the accepted limits (< 15 %) throughout the validation period. The present method is useful for determination of plasma concentrations of ondansetron during human pharmacokinetic studies. PMID- 15553105 TI - Clinical study of the therapeutic efficacy and safety of emedastine difumarate versus terfenadine in the treatment of seasonal allergic rhinitis. AB - OBJECTIVE: Emedastine is a new H1-receptor antagonist endowed with potent and selective antihistamine activity. The aim of this study was to evaluate the therapeutic efficacy and tolerability of emedastine difumarate (CAS 87233-62-3) in Caucasian patients in the treatment of seasonal allergic rhinitis as compared to terfenadine (CAS 50679-08-8). METHODS: A total of 130 patients suffering from grass pollen allergic rhinitis were randomly assigned to 14 days treatment with either emedastine difumarate (2 mg b.i.d.) or terfenadine (60 mg b.i.d.) in a double-blind, randomised, crossover design. Primary efficacy parameter was a Total Severity Symptom Score, including among symptoms nasal congestion, sneezing, rhinorrhea, nasal/throat/palate itching, eye itching and lacrimation. Safety was assessed on routine laboratory tests and recording vital signs and adverse events (AEs). RESULTS: Emedastine 2 mg b.i.d. was significantly more effective than terfenadine 60 mg b.i.d. in reducing Total Symptom Severity Score (p = 0.0258). This statistical significant difference was also obtained in controlling sneezing and rhinorrhea (p = 0.003). Moreover, both the physician and patients indicated emedastine as the preferred therapy (p < 0.01). Forty-seven drug related AEs were reported for emedastine (= 51.07 %) and 53 for terfenadine (64.15 %), most of them involving the CNS. CONCLUSION: The results of study show that emedastine difumarate is more effective than terfenadine in the symptomatic management of seasonal allergic rhinitis and is particularly active in controlling the main nasal symptoms, such as sneezing and rhinorrhea; it is safe and well tolerated in this therapeutic indication, while related AEs are less if compared to those displayed by terfenadine. PMID- 15553106 TI - Clinical study of the therapeutic efficacy and safety of emedastine difumarate versus cetirizine in the treatment of seasonal allergic rhinitis. AB - OBJECTIVE: The therapeutic efficacy and tolerability of emedastine difumarate (CAS 87233-62-3) in male and female Caucasian patients with seasonal allergic rhinitis as compared to cetirizine (CAS 83881-51-0) was evaluated. METHODS: The study was designed as a double-blind, randomised, parallel groups comparison of two antihistamines administered by oral route (emedastine 4 mg o.d. versus cetirizine 10 mg o.d.) in a population of 120 patients suffering from grass pollen allergic rhinitis. The duration of the treatment period was 14 days. Primary efficacy variable was a total symptoms score (including among symptoms nasal congestion, sneezing, rhinorrhea, nasal/throat/palate itching, eye itching and lacrimation) evaluated after 14 days of treatment vs. baseline value. Safety was assessed on routine laboratory assays and recording vital signs and adverse events (AEs). RESULTS: The between-group difference in primary efficacy variable averaged over the 2-week treatment period was not statistically significant. Results clearly showed that no significant difference exists between the two treatments as far as total symptoms score evaluated after 14 days of treatment vs. baseline values are concerned. Therefore, the efficacy profiles of the study medications are overlapping. The pattern and incidence of AEs was similar in both treatment groups. The most frequent AEs with both compounds were related to the CNS, headache being the most reported one. In particular, this study seems to disclose a slighter tendency to drowsiness with emedastine than with cetirizine. CONCLUSIONS: Both drugs under investigation in this trial appear to be effective for relieving the symptoms of seasonal allergic rhinitis in Caucasian adult patients. The results demonstrate that emedastine 4 mg o.d. is comparable in efficacy to cetirizine 10 mg once daily in the symptomatic management of seasonal allergic rhinitis. Moreover, based on the results of this study, emedastine can be considered a safe and well-tolerated drug and its safety profile seems to resemble that of cetirizine. PMID- 15553107 TI - Analgesic effects of low-dose intravenous orphenadrine in the state of capsaicin hyperalgesia. A randomised, placebo-controlled, double-blind cross-over study using laser somatosensory evoked potentials obtained from capsaicin-irritated skin in healthy volunteers. AB - The present investigation aimed to elucidate the analgesic efficacy of 30 mg of intravenous orphenadrine citrate (CAS 4682-36-4) in a human pain model. Eighteen healthy female and male subjects were enrolled and received single infusions of 30 mg orphenadrine citrate and matching placebo in two periods which were separated by a 1 week washout period. The study was designed as a randomised, double-blind, placebo-controlled, two-period, cross-over trial. The intended neurogenic inflammation and hyperalgesia were induced by topical, occlusive application of 1% capsaicin solution (INCI: Capsicum frutescens, containing capsaicinoides from Capsicum annuum annuum, CAS 84603-55-4) for 30 min on defined skin areas of the back. The pain response to CO2 laser pulses applied to the capsaicin pre-treated skin was measured by event related Vertex-EEG recordings. This technique allowed studying the influence of orphenadrine citrate on the (central) P2-component and the (peripheral) Ni-component of the pain response (LSEP). Both, orphenadrine citrate and placebo were given as intravenous infusions over 60 min. Orphenadrine citrate exerted a significant reduction in central and peripheral components of the pain response when compared to placebo. The effect on the central component was highly significant and more pronounced than the peripheral effect of the drug. The analgesic effect developed fast, was already present during infusion, was ongoing, and exceeded the observational period of 4 h after start of infusion. In summary, orphenadrine citrate was able to exert an analgesic/anti-hyperalgesic effect in a low-dose paradigm (30 mg dose) which was predominantly due to central/spinal mechanisms in this capsaicin model with laser somatosensory evoked potentials. PMID- 15553108 TI - Bioequivalence study of levothyroxine tablets compared to reference tablets and an oral solution. AB - The study was designed to evaluate the bioequivalence of three levothyroxine sodium (CAS 51-48-9) formulations, i.e. a test and a reference tablet and an oral solution. A bioequivalence study was carried out in 25 healthy volunteers, who were administered a single dose of 600 microg levothyroxine in the form of the test formulation (levothyroxine sodium tablets 200 microg; Eferox), the originator product, and an oral solution. The trial was performed in one study center according to an open, randomized, three-way cross-over design with wash out periods of 35 days between administration. Blood samples were taken up to 48 h post dose, the plasma was separated and the concentrations of levothyroxine and triiodothyronine were determined by radioimmunoassay with I125 labeling method. The levothyroxine mean Cmax were 112.0+/-17.3 ng/ml, 113.4+/-18.5 ng/ ml and 111.3+/-15.1 ng/ml, while the mean AUC0-24 were 2263.7+/-332.8 ng x h/ ml, 2307.3+/-351.3 ng x h/ml and 2286.1+/-331.0 ng x h/ml for the test and reference tablets as well as for the oral solution, respectively. No significant differences were found of principal pharmacokinetic parameters between the studied formulations. The 90%-confidence interval for the primary target parameters, intra-individual ratios of AUC0-24 and Cmax of levothyroxine were within the acceptance ranges for bioequivalence trials, i.e. AUC0-24 0.954-1.016 and 0.966-1.011 as well as Cmax 0.948-1.027 and 0.968-1.032 for test tablets versus reference tablets and the oral solution, respectively. Similar results were observed for triiodothyronine. In the light of the present study it can be concluded that the levothyroxine test tablet is bioequivalent to the reference formulation in respect of extent and rate of absorption. The results of the present trial confirm the findings of a previous study, performed under steady state conditions with Eferox tablets 100 microg in patients without thyroid function. PMID- 15553109 TI - Analysis of pooled plasma samples as a predictor for proving bioequivalence of drugs with a long elimination half-life. The example of phenprocoumon. AB - The results of pooled plasma analysis in a bioequivalence trial provide information comparable with that of the mean concentration vs. time curves for each formulation. Although it seems feasible that pool plasma analysis will have similar or even greater advantages in cases of bioequivalence trials with a parallel-group design, no such data has been published in the literature probably due to the limited number of such trials. The present study was designed with the aim to investigate the prediction value of pool plasma analysis in a bioequivalence trial with phenprocoumon (CAS 435-972). The study was performed as a monocentric, randomized, open clinical trial in two parallel groups of healthy male volunteers (31 per group), all of which received a single oral dose of 3 mg phenprocoumon. Serial blood samples were drawn for the pharmacokinetic analysis pre-dose, and 0.33, 0.67, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 12, 24, 48, 96, 144, 192, and 240 h after drug administration. Pool plasma was prepared for each sampling point and each formulation. Drug concentrations were measured by means of an HPLC method. A comparison between the pool plasma results and the results of individual analysis revealed a very good correspondence regarding the parameters AUC, Cmax and t(max). The present trial demonstrates that the method of time-wise pooling provides reliable information not only in cross-over trials but also in trials with parallel groups of volunteers. PMID- 15553110 TI - High resolution X-ray structure and potent anti-HIV activity of recombinant dianthin antiviral protein. AB - Dianthin antiviral protein (DAP) is a naturally occurring antiviral protein from the leaves of carnation (Dianthus caryophyllus) capable of depurinating HIV-1 RNA and inhibiting HIV-1 replication in human peripheral blood mononuclear cells. Escherichia coli-derived recombinant DAP (rDAP, amino acids 1-254) was purified to homogeneity for structural and functional studies. In the following paper the X-ray crystal structure of rDAP as well as its complexes with cyclic AMP and adenyl-guanosine (ApG) as substrate analogs at 1.7 A resolution are reported. Molecular modeling studies of the interactions of DAP and the structurally similar pokeweed antiviral protein (PAP) with a single-stranded RNA heptamer predicted a more potent anti-HIV activity for rDAP due to its unique surface topology and more favorable charge distribution in its 20 A-long RNA binding active center cleft. In accordance with the predictions of the modeling studies, rDAP was more potent than rPAP in depurinating HIV-1 RNA. To the knowledge of the authors, this is the first structural and functional characterization of recombinant DAP. PMID- 15553111 TI - UEMS charter on training of medical specialists in the EU--the new neurosurgical training charter. PMID- 15553112 TI - Neurosurgical training programme director position and responsibilities. PMID- 15553113 TI - Teaching staff. PMID- 15553114 TI - The ideal neurosurgical training curriculum. PMID- 15553115 TI - Internal regulations and general guidelines of a neurosurgical department and training programme. AB - It is strongly recommended that general guidelines and internal regulations be laid down in written form by the department. The guidelines regulate the general workflow and related trainee duties and responsibilities, education and research, special procedures for particular diseases and conditions (standards of care). Regarding general workflow of the department, the following items should be laid down once in written form: time table of department, working hours, admission procedures, medical records directives, responsibilities on wards and in the emergency unit, patient information, discharge procedures, outpatient consultations, call schedule and on-call duty plan, as well as week-end and holiday regulations for rounds on wards and ICU. Regarding education and research, the following items should be written down: conferences and teaching rounds, policies on presentations and publications, policies regarding meetings and courses, research leave and vacation plan. The written definition of standards of care is still considered optional in neurosurgery at the present time. PMID- 15553116 TI - Principles of teaching in a structured training programme, the rotations, and the surgical training plan. AB - The present paper is divided into two parts. In the first part it summarises the essentials of transfer of knowledge and personality from trainer to trainee. The training capacity of a programme should be identified first since the number of residents in training has implications on the structure of the programme. The capacity is limited by the number of surgical "resident cases", which should at least amount to 70 per year and resident. For the future, more emphasis should be laid on the acquirement of methods of self-teaching and continuous self education, in order to provide the trainees with the ability to cope with changes during their later career more easily. In part two the organisation of rotations as well as the structured surgical training plan are discussed. Teaching can be organised either as a gradual exposure to more and more complex procedures or as a sequence of speciality rotations. Structured teaching of theoretical contents should accompany practical teaching and in academic programmes a scientific rotation should be integrated. The ongoing subspecialisation is currently exceeding the limits of complete coverage during a six-year-programme. PMID- 15553117 TI - A structured neurosurgical training plan and the neurosurgical logbook in the UK. PMID- 15553118 TI - The European Neurosurgical Log-Book (UEMS/EANS). PMID- 15553119 TI - Morbidity & mortality conferences--how can we do it? PMID- 15553120 TI - Problem-based learning in residency training and the tutorial process for training and education in neurosurgery. PMID- 15553121 TI - Periodic progress evaluation. PMID- 15553122 TI - Research rotation in a trainee's curriculum. PMID- 15553123 TI - The accreditation of a training programme. PMID- 15553124 TI - The European examination--its present status and potential development. PMID- 15553125 TI - Neurosurgical subspecialization: pros and cons. PMID- 15553126 TI - Comprehensive two-dimensional chromatography in food analysis. AB - Comprehensive two-dimensional (2D) chromatographic techniques can be considered innovative methods, only quite recently developed. Since their introduction to the chromatographic community, these techniques have been used in several fields and have gained an excellent reputation as valuable and powerful analytical tools. The revolutionary aspect of comprehensive multidimensional (MD) techniques, in respect to classical MD chromatography, is that the entire sample is subjected to the 2D advantage. The resulting unprecedented separating capacity makes these approaches prime choices when analysts are challenged with highly complex mixtures. Furthermore, in the case of automated systems, instrumental analysis times are roughly the same as in monodimensional applications. The present review reports various comprehensive chromatographic applications on different food matrices. The GC x GC section highlights two fundamental aspects for component separation/identification: the exceptional peak capacity and the formation of group types on the 2D space plane. The LC x LC section reports the employment in food analysis of a recently developed multidimensional normal-phase (NP)-reversed-phase (RP) high performance liquid chromatography (HPLC) system. Also reported are comprehensive LC x GC and packed column supercritical fluid chromatography (pSFC x pSFC) applications in this field. PMID- 15553127 TI - Volatile compounds in virgin olive oil: occurrence and their relationship with the quality. AB - The stimulation of the human sensory receptors by volatile compounds present in virgin olive oils gives rise to the sensory attributes that describe its delicate and fragrant aroma. The composition of the volatile compounds and their biogenesis is briefly illustrated. Analytical methodologies for evaluating the volatile fraction and the sensory properties of virgin olive oils are elucidated. Compounds responsible for typical flavours are examined and the influence of the main factors on the composition of volatile compounds is discussed. The origin of off-flavours are also described and the consequent changes of volatile composition and of sensory characteristics are analysed. The relationships between volatile compounds and sensory attributes are discussed. PMID- 15553128 TI - Evaluation of two gas chromatography-olfactometry methods: the detection frequency and perceived intensity method. AB - Two gas chromatography-olfactometry methods were evaluated in terms of repeatability, range of sensitivity and discriminating properties. Six volatile flavour compounds at various concentration levels were analysed by a panel of eight assessors using the detection frequency method and the perceived intensity method. The coefficient of variance, averaged over the individual compounds for three replicate samples, was 16% for the detection frequency method and 28% for the intensity method. The average correlation coefficient of the individual compounds with concentration was 0.93 (range 0.88-0.99) for the intensities. They were slightly higher than those for the detection frequencies (0.91, range 0.81 0.97). The detection frequency method was more accurate in terms of repeatability, and the intensity method was more accurate with regard to discrimination between concentration levels. The range of sensitivity was similar for both methods. PMID- 15553129 TI - Correlating sensory attributes to gas chromatography-mass spectrometry profiles and e-nose responses using partial least squares regression analysis. AB - Sensory scores for 15 attributes identified in soy sauce aroma by quantitative descriptive analysis were correlated with purge and trap gas chromatography-mass spectrometry (GC-MS) profiles and electronic nose (e-nose) responses using partial least squares (PLS) regression analysis. Highly predictive PLS models were obtained for every attribute based on whole GC-MS profiles. However, the predictability has been greatly improved in the models calculated from 20 selected peaks that showed higher contribution to each attribute in the first PLS analysis. Contrarily, except for alcoholic and fishy notes, predictability of PLS models calculated from e-nose responses was poor. The correlation between GC-MS profiles and e-nose responses was unsatisfactory due to high similarity in sensor responses. PMID- 15553130 TI - Comprehensive two-dimensional gas chromatography of complex samples by using a 'reversed-type' column combination: application to food analysis. AB - The practicability and potential of a non-orthogonal approach in comprehensive two-dimensional gas chromatography (GC x GC) were studied and compared to those of the orthogonal approach for two different complex matrices, and using conventional flame ionisation (FID) and time-of-flight mass spectrometry (ToF MS) detection. The separation of a diesel oil showed that the non-orthogonal approach also provides interesting, but completely reversed, ordered structures. For the more extensively studied flavour analysis in food samples, improved peak shapes and, also, different types of ordered structures and retention behaviour, and improved detectability for polar compounds make the two approaches complementary to each other. As a consequence, identification and/or determination of targets and/or unknowns can be performed more reliably. Analytical performance (close to three-order linearity; LODs, 2-30 pg injected in most cases; R.S.D.s, 1-6% (n = 6)) was fully satisfactory. PMID- 15553131 TI - Analysis of roasted coffee bean volatiles by using comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry. AB - The volatile headspace from two coffee bean varieties, namely Arabica (Coffea arabica) and Robusta (Coffea canephora ex Froehner), were sampled by using solid phase microextraction (SPME), and then analysed with comprehensive two dimensional gas chromatography interfaced to a time-of-flight mass spectrometer (GC x GC-TOFMS). Two distinct column set combinations were investigated--an apolar-polar and polar-apolar configuration--and the separation achieved from each set was compared. Results were compared with a previous literature report for coffee analysed by GC x GC-FID, using an analogous polar-apolar column set combination, where authentic standards were used to confirm the position of the selected components in the 2D separation space. The present study provides independent mass spectral confirmation of component identity, and demonstrates that the relative, structured position of these components is comparable in the two experiments. Total ion current (TIC) chromatograms were processed using ChromaTOF automated data processing software. It was necessary to restrict the number of processed peaks to 1000 (S/N > 100), which required approximately 8 h for processing. Extracted ion chromatograms were generated using prominent fragment ions, and unique masses, to aid in analyte identification process, and was particularly useful in instances of component peak overlap, and for the identification of pyrazine analytes (e.g. 44, 88, 122 u). Semi-quantitative analysis was restricted to the 44 selcted components; however, the omission of peaks with S/N < 100, limiting the processed peaks to 1000, reduced the semi quantitative application of the GC x GC-TOFMS method developed. Finally, results gained from GC x GC-TOFMS and GC x GC-qMS analyses were comparable with respect to spectral similarity assignments for the 44 target analytes. PMID- 15553132 TI - Essential oil yield and quality of methyl eugenol rich Ocimum tenuiflorum L.f. (syn. O. sanctum L.) grown in south India as influenced by method of harvest. AB - A field experiment carried out during 2001--2002 under semi-arid conditions of Hyderabad, India investigated the effect of three different methods of harvesting at full bloom stage, on essential oil yield and quality of methyl eugenol rich sacred/holy basil (Ocimum tenuiflorum L.f.; Lamiaceae). The harvest methods were: harvesting of primary branches, secondary branches and shoot biomass cut at 30 cm above ground level. Four harvests at 102, 192, 287 and 360 days after transplanting of the crop were taken in 1 year in each method of harvest. Harvesting of secondary branches led to maximum plant height and number of secondary branches per plant compared to harvesting of primary branches or shoot biomass cut at 30 cm above ground during second, third and fourth harvests. On the contrary, secondary branch harvest gave least biomass yield in all the four harvests. But due to higher essential oil content, secondary branch harvest gave 25.2 and 15.4% higher total (sum total of all four harvests) essential oil yield (kg/ha per year) over primary branches and shoot biomass cut at 30 cm above ground methods of harvesting, respectively. A similar treatment difference was observed in respect of oil composition studied in the first harvest. Harvesting shoot biomass at 30 cm above ground produced oil containing highest amount of methyl eugenol. The content of methyl eugenol decreased in the order of shoot biomass cut at 30 cm above ground > primary branch > secondary branch treatments. A reverse trend was observed, however, in respect of (E)-cinnamyl acetate, eugenol and beta-elemene constituents of the oil. Little variability was, however, observed among the treatments in respect of 24 other constituents of the oils. PMID- 15553133 TI - Comparison of the volatile constituents of Artemisia capillaris from different locations by gas chromatography-mass spectrometry and projection method. AB - The volatile chemical constituents of Artemisia capillaries (an important traditional Chinese medicine) were determined by gas chromatography-mass spectrometry (GC-MS) and sub-window factor analysis (SFA). Seventy-five components were separated and 43 of them were qualitatively and quantitatively determined, which represented about 89.03% of the total content. This profile was then used to identify and assess the consistency of the herb by using an orthogonal projection method. Four different sources of A. capillaries were analyzed and compared with each other. Among the components determined, there were 51 components coexisting in all samples although the relative peak areas of a few showed variations. It is the first time to apply orthogonal projection method to the comparison of different samples, and it reduces the burden of qualitative analysis as well as the subjectivity. The results showed a fair consistency in their GC-MS fingerprint. A. capillaris was distinguished from Artemisia sacrorum L., a possible substitute in traditional Chinese medicine by comparing the fingerprints with each other. PMID- 15553134 TI - On-line reversed-phase liquid chromatography-gas chromatography coupled to mass spectrometry for enantiomeric analysis of chiral compounds in fruit beverages. AB - A method based on the on-line coupling of reversed phase liquid chromatography with gas chromatography/mass spectrometry (RPLC-GC-MS) for the chiral evaluation of characteristic constituents of fruit beverage aroma was investigated. The consideration of a variety of parameters involved in the transfer step allowed to achieve relative standard deviations ranging from 0.4 to 10% in most cases and detection limits from 0.2 to 2.5 mg/l. By applying the developed method to fruit beverages, racemic mixtures of ethyl 2-methylbutanoate and gamma-nonalactone were found. This fact suggests the eventual addition of artificial aromas. The method proposed in the present work can be useful to assess reliably the authenticity of aqueous samples, such as fruit beverages. PMID- 15553135 TI - Natural variability of the enantiomeric composition of bioactive chiral terpenes in Mentha piperita. AB - The variability of the enantiomeric distribution of biologically active chiral terpenes in Mentha piperita plants from different geographical origins was evaluated by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). The optimisation of some parameters (i.e. exposure temperature, extraction time and type of fibre) affecting SPME-extraction enabled relative standard deviations ranging from 4 to 13% to be achieved. The use of two different chiral stationary phases allowed to separate the identified chiral terpenes into their corresponding enantiomers as well as to verify the enantiomeric excesses of those compounds which were enantiomerically resolved on both phases. For all chiral terpenes, the enantiomeric composition varied within a very narrow range all over the samples. Consequently, it may be stated that the enantiomeric composition of chiral terpenes in Mentha piperita appears to be independent of the geographical origin of the plant and, thus, any alteration in the characteristic value may be related to an adulteration or inadequate sample handling. These results support the usefulness of the enantiomeric composition of bioactive chiral terpenes in Mentha piperita in authenticity studies. PMID- 15553136 TI - Extraction and analysis of phenolics in food. AB - Phenolics are ubiquitous compounds found in all plants as their secondary metabolites. These include simple phenols, hydroxybenzoic acid and cinnamic acid derivatives, flavonoids, coumarines and tannins, among others. The extraction of phenolics from source materials is the first step involved in their analysis. While chemical methods are used for determination of total content of phenolics, chromatographic and spectrometric analyses are employed for identification and quantification of individual compounds present. This paper provides a summary of background information and methodologies used for the analysis of phenolics in foods and nutraceuticals. PMID- 15553137 TI - Health and sensory properties of virgin olive oil hydrophilic phenols: agronomic and technological aspects of production that affect their occurrence in the oil. AB - Hydrophilic phenols are the most abundant natural antioxidants of virgin olive oil (VOO), in which, however, tocopherols and carotenes are also present. The prevalent classes of hydrophilic phenols found in VOO are phenolic alcohols, phenolic acids, flavonoids, lignans and secoiridoids. Secoiridoids, that include aglycon derivatives of oleuropein, demethyloleuropein and ligstroside, that are present in olive fruit, are the most abundant phenolic antioxidants of VOO. The sensory and healthy proprieties of VOO hydrophilic phenols as well as the agronomic and technological parameters that affect their concentration in the oil are discussed in this paper. PMID- 15553138 TI - Anthocyanins in cereals. AB - The anthocyanic composition of some pigmented cereals is still not well established, neither in relation to some of their components, nor from the quantitative point of view. Nonetheless, the use of analytical techniques, such as diode array spectroscopy and mass spectrometry (MS, PDMS, MALDI) coupled or not to liquid chromatography, are permitting, in recent years, the confirmation of the structure of some of the principal anthocyanins and a knowledge of those which are present in minor proportion. In this article, firstly, a review of the principal methods of analysis of anthocyanins is made. This is followed by a review of the most significant advances achieved in the last years in the field of the identification and quantification of these pigments in cereals and the present uses of the commercial extracts of anthocyanins obtained from these sources and the perspectives for their use is included. PMID- 15553139 TI - Analysis of biologically active compounds in potatoes (Solanum tuberosum), tomatoes (Lycopersicon esculentum), and jimson weed (Datura stramonium) seeds. AB - Potatoes and tomatoes, members of the Solanaceae plant family, serve as major, inexpensive low-fat food sources providing for energy, high-quality protein, fiber, vitamins, pigments, as well as other nutrients. These crops also produce biologically active secondary metabolites, which may have both adverse and beneficial effects in the diet. This limited overview, based largely on our studies with the aid of HPLC, TLC, ELISA, GC-MS, and UV spectroscopy, covers analytical aspects of two major potato trisaccharide glycoalkaloids, alpha chaconine and alpha-solanine, and their hydrolysis products (metabolites) with two, one, and zero carbohydrate groups; the potato water-soluble nortropane alkaloids calystegine A3 and B2; the principal potato polyphenolic compound chlorogenic acid; potato inhibitors of digestive enzymes; the tomato tetrasaccharide glycoalkaloids dehydrotomatine and alpha-tomatine and hydrolysis products; the tomato pigments beta-carotene, lycopene, and chlorophyll; and the anticholinergic alkaloids atropine and scopolamine present in Datura stramonium (jimson weed) seeds that contaminate grain and animal feed. Related studies by other investigators are also mentioned. Accurate analytical methods for these food ingredients help assure the consumer of eating a good-quality and safe diet. PMID- 15553140 TI - Chromatographic analysis of alkylresorcinols and their metabolites. AB - Alkylresorcinols (AR) are phenolic lipids present in high amounts in the bran fraction of wheat and rye. AR are of scientific interest as bioactive compounds, as markers for the presence of wholegrain or bran fractions of wheat and rye in food products, and as possible biomarkers of intake for wholegrain wheat and rye. This review discusses their extraction from cereal grains, food and biological fluids, and the various chromatographic methods used in their analysis. PMID- 15553141 TI - High-performance anion-exchange chromatography coupled with pulsed amperometric detection and capillary zone electrophoresis with indirect ultra violet detection as powerful tools to evaluate prebiotic properties of fructooligosaccharides and inulin. AB - Fructooligosaccharides (FOS) and inulin are food grade non-digestible carbohydrates that exert beneficial nutritional effect. This paper describes the suitability of high-performance anion-exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) and capillary zone electrophoresis (CZE) to evaluate fermentation properties of FOS and inulin in pure Bifidobacterium cultures; and to study their effects on faecal cultures (microbial population and short-chain fatty acids). Prebiotic effectiveness of FOS and inulin of different degrees of polymerization was evaluated monitoring the changes in their molecular weight distribution during the in vitro growth of selected Bifidobacterium strains. The qualitative analysis of the residual soluble oligosaccharides or polysaccharides from Raftilose Synergy, Raftiline HP and Raftilose P95 was carried out by HPAEC-PAD, using a CarboPac PA 100 column and an appositely optimized gradient elution program. Under the optimized gradient elution conditions, glucose, fructose, sucrose were resolved from each other and from fructans with a DP ranging from 3 (1-kestose) to 60. The chromatographic profiles of the spent broths pointed out that almost every strain presented a different capability to ferment fructan chains of variable DP, indicating wide strain to strain differences. To explore the prebiotic effect of FOS and inulin, related to of short chain fatty acids (SCFAs) accumulation in faecal cultures due to fermentative metabolism of intestinal microflora, analysis of SCFAs, acetic and lactic acid was achieved by co-electroosmotic capillary electrophoresis, where the electrophoretic mobility of the anionic analytes and electroosmotic flow (EOF) were similarly directed. Moreover, the use of UV detection for the analyses of our organic anions required a running electrolyte which allowed indirect detection. The optimization of the capillary electrophoretic conditions was carried out by applying a chemometric study based on the use of the experimental design, the effects of three parameters, i.e. temperature, voltage and percentage of methanol added to the background electrolyte were investigated. PMID- 15553142 TI - Development of a rapid method based on solid-phase extraction and liquid chromatography with ultraviolet absorbance detection for the determination of polyphenols in alcohol-free beers. AB - An analytical method based on solid-phase extraction (SPE) and followed by liquid chromatographic separation and ultraviolet detection (HPLC-UV) is proposed for the determination of 10 phenolic compounds which participate on beer stability and sensory properties in alcohol-free beers. Acetonitrile was found to be the most appropriate solvent for the elution of polyphenolic compounds adsorbed on C18 cartridges. The performance of the method was assessed by the evaluation of parameters such as absolute recovery (generally higher than 60%), repeatability (lower than 10%), linearity (r2 higher than 0.993) and limits of quantitation (ranging from 1 to 37 microg/L); no matrix effects were observed. The polyphenol content of different Spanish alcohol-free beers is presented. Five phenolic compounds such as protocatechuic, p-coumaric, ferulic, caffeic acids, and (+) catechin were identified at levels lower than 10 mg/L. PMID- 15553143 TI - Characterisation of flavonols in broccoli (Brassica oleracea L. var. italica) by liquid chromatography-uV diode-array detection-electrospray ionisation mass spectrometry. AB - The flavonoid composition of broccoli inflorescences has been studied by LC/UV DAD/ESI-MSn. A large number of hydroxycinnamic acid esters of kaempferol and quercetin glucosides has been characterised. The structures of the flavonoid glycosides were analysed after alkaline hydrolysis, and were identified as 3 sophoroside/sophorotrioside-7-glucoside/sophoroside of kaempferol, quercetin and isorhamnetin (this last found in trace amount). These complex quercetin and isorhamnetin glucosides have not been previously characterised in nature. In addition, several less complex glucosides based on the same aglycones have been identified. The effect of sugar substitution and acylation on chromatographic mobility and ESI ionisation and fragmentation are discussed. PMID- 15553144 TI - Determination of anthocyanins in wine by direct injection liquid chromatography diode array detection-mass spectrometry and classification of wines using discriminant analysis. AB - A rapid HPLC-diode array detection (DAD) method was developed for the routine analysis of 16 anthocyanins in wine. Direct injection of filtered wine samples followed by selective detection at 520 nm allowed quantitation of these compounds in red wines. The method was linear for malvidin-3-glucoside over the range 5 250ppm, and the limit of detection for this compound was 0.18 ppm. A volatile mobile phase is used, which enables hyphenation to mass spectrometry (MS). With HPLC-MS, a total of 44 pigments could be identified in South African wines. Obtained mass spectra are discussed for a series of representative wine constituents and results are compared with literature references. An attempt was made to differentiate between different cultivars according to the anthocyanin content using stepwise forward linear discriminant analysis (LDA). PMID- 15553145 TI - Liquid chromatographic-mass spectrometric analysis of anthocyanin composition of dark blue bee pollen from Echium plantagineum. AB - Dark blue bee pollen samples from pollinic type Echium plantegineum were analysed in order to identify and quantify their anthocyanin pigments. Five samples were collected from different apicultural Spanish regions and the anthocyanin composition was determined by HPLC with diode array and MS detection. Eight different pigments were identified, the principal anthocyanin being petunidin-3-O rutinoside. The other pigments found were delphinidin, cyanidin and petunidin-3-O glucoside; delphinidin, cyanidin, peonidin and malvidin-3-O-rutinoside and cyanidin-3-(6"-malonylglucoside). The anthocyanin content ranged from 45 to 80 mg/100 g of blue pollen, which could represent a significant source of phytochemicals. Minor variations in the anthocyanin profiles were found, which could be explained by the geographical differences between collection regions. PMID- 15553146 TI - Liquid chromatography-mass spectrometry identification of anthocyanins of isla oca (Oxalis tuberosa, Mol.) tubers. AB - High-performance liquid chromatography (HPLC)-diode array detection (DAD)-mass spectrometry (MS) techniques have been successfully employed in the identification of the anthocyanins of the coloured tubers of isla oca (Oxalis tuberosa), the second most cultivated tuber in the Andean region. Tubers underwent a pre-treatment step in order to inhibit enzymatic reactions and to obtain a stable powder or "concentrate". This concentrate was dissolved, purified and then analysed. Eight different compounds were found. The major peaks were malvidin glucosides (malvidin 3-O-glucoside and 3,5-O-diglucoside). The rest of the peaks were 3,5-O-diglucosides of petunidin and peonidin, and 3-O-glucosides of delphinidin, petunidin and peonidin. Only malvidin 3-O-acetylglucoside-5-O glucoside was found as an acylated anthocyanin. PMID- 15553147 TI - Determination of pigments in vegetables. AB - Plant pigments are responsible for the shining color of plant tissues. They are also found in animal tissues and, eventually in transformed food products as additives. These pigments have an important impact on the commercial value of products, because the colors establish the first contact with the consumer. In addition plant pigments may have an influence on the health of the consumers. Pigments are labile: they can be easily altered, and even destroyed. Analytical processes have been developed to determine pigment composition. The aim of this paper is to provide a brief overview of these methods. PMID- 15553148 TI - Tocopherol measurement in edible products of vegetable origin. AB - Since natural antioxidants present increasing interest for food industry due to their beneficial effect on health, new potential sources have been screened among edible aromatic plants and a microalgae, Spirulina platensis. The determination was performed after optimising a previously validated method, because important differences have been found among values described in literature for tocopherol content in products of vegetable origin. Values obtained ranged from 3.42 mg alpha-tocopherol/100 g of dill to 132.2 mg/100 g of fresh bay and from 0.14 mg gamma-tocopherol/100 g of spearmint to 3.45 mg/100 g of parsley. In all cases results were calculated from fresh leaves. Preliminary experiments were developed with bay (Laurus nobilis) plant to devise the supercritical fluid extraction of tocopherols, generating environmentally friendly processes to selectively extract fractions enriched with antioxidant compounds while removing fractions corresponding to essential oils, that is, those that correspond to the characteristic aroma of the plants. Another striking result has been the tocopherol content in the microalgae, 1.3 mg alpha-tocopherol/100 g of dried commercial spirulina, which do not justify the supposed source of antioxidant vitamins. Results suggest the need of more reliable determinations of tocopherols in vegetable sources to be included in databases. PMID- 15553149 TI - Evaluation of a methylation procedure to determine cyclopropenoids fatty acids from Sterculia striata St. Hil. Et Nauds seed oil. AB - Cyclopropenoids fatty acids (CPFA) from Sterculia striata seed oil were characterized by gas chromatography/mass spectrometry (GC/MS) and quantified by gas chromatography-flame ionization detector (GC-FID) after derivation to fatty acid methyl esters using a cold base-catalyzed procedure. 1H nuclear magnetic resonance (NMR) analysis were done in oil and fatty acid methyl esters derivatives to quantify CPFA and verify artifacts formation during the base catalyzed reaction. Similar quantities of CPFA were found in S. striata and Sterculia foetida seed oils before and after a base-catalyzed methylation by NMR analysis, with no artifact formation. These results were compatible with those obtained by GC-FID analysis. Transmethylation with KOH in methanol was an appropriated method to prepare cyclopropenoids fatty acids methyl esters and quantify them by GC and NMR analysis. PMID- 15553150 TI - Liquid chromatographic analysis of milk phospholipids with on-line pre concentration. AB - Two methods have been developed for the analysis of bovine milk phospholipid (PL) classes by NP-HPLC with evaporative light scattering detection. In the first method, a PVA-Sil guard column was used for the rapid determination of the major milk PL, phosphatidylethathanolamine (PE), phosphatidylcholine (PC) and sphingomyelin (SM). In the second method, the guard column was used to pre concentrate the PL, which were then transferred on-line onto a PVA-Sil analytical column by the use of column switching valves. This enabled separation of complete milk PL, including phosphatidylinositol (PI), phosphatidylserine (PS) and lysophosphatidylcholine (LPC). PMID- 15553151 TI - Determination of mono- and di-acylglycerols in milk lipids. AB - A simple and rapid method based on solid phase extraction and gas chromatography has been developed for the determination of monoacylglycerols (MAG) and diacylglycerols (DAG) at low-concentration levels typically found in milk and dairy ingredients. The method enabled measurement of individual milk MAG (including isomeric forms) with the exception of glycerol, monobutyrate. The DAG were separated and quantified as groups according to their carbon numbers. However, it was possible to identify the major DAG components within a group. The minimum detection limits for MAG and DAG were in the range of 5-8 and 10-17 microg/ml, respectively. The corresponding R.S.D. values were 1.7-3.9 and 0.2 9.9%, respectively. C8-C18 MAG and C26-C36 DAG were present in the lipids extracted from whole milk, anhydrous milk fat and buttermilk. The concentrations of MAG and DAG in buttermilk were, respectively, thirteen- and three-folds higher than that in whole milk or anhydrous milk fat. In dairy lipids, 1,2(2,3)-DAG isomers predominated almost to the exclusion of 1,3-isomers. PMID- 15553152 TI - Improved liquid chromatographic method for determination of carotenoids in Taiwanese mango (Mangifera indica L.). AB - An HPLC method was developed to determine the various carotenoids in Taiwanese mango (Mangifera indica L.). Initially, the peel and seed of mangoes were removed, the pulps were cut into pieces, freeze-dried, ground into powder, extracted and subjected to HPLC analysis. A mobile phase of methanol-isopropanol (99:1, v/v) (A) and methylene chloride (100%) (B) with the following gradient elution was developed: 100% A and 0% B in the beginning, maintained for 15 min, decreased to 70% A in 45 min, maintained for 15 min and returned to 100% A in 65 min. A total of 25 carotenoids were resolved within 53 min by using a C-30 column with flow rate at 1 mL/min and detection at 450 nm. alpha-Carotene was used as an internal standard to quantify all the carotenoids. All-trans-beta-carotene was present in largest amount (29.34 microg/g), followed by cis isomers of beta carotene (9.86 microg/g), violaxanthin and its cis isomers (6.40 microg/g), neochrome (5.03 microg/g), luteoxanthin (3.6 microg/g), neoxanthin and its cis isomers (1.88 microg/g), zeaxanthin (1.16 microg/g) and 9- or 9'-cis-lutein (0.78 microg/g). PMID- 15553153 TI - Identification and formation of angiotensin-converting enzyme-inhibitory peptides in Manchego cheese by high-performance liquid chromatography-tandem mass spectrometry. AB - A total of 75 peptides included in the fraction with molecular mass below 3000 from an 8-month-old Manchego cheese could be identified using HPLC coupled on line to an ion trap mass spectrometer. Some previously described peptides with antihypertensive and/or angiotensin-converting enzyme (ACE)-inhibitory activity were detected. The formation of five active sequences was followed during cheese ripening in four different batches of Manchego cheese. Two experimental batches of Manchego cheese elaborated with selected bacterial strains with the aim of improve the organoleptic characteristics demonstrated also a good performance in the formation of peptides with ACE-inhibitory activity during cheese ripening. PMID- 15553154 TI - Quantitative determination of alpha(s2)- and alpha(s1)-casein in goat's milk with different genotypes by capillary electrophoresis. AB - A capillary electrophoresis (CE) method has been applied for the quantitative determination of alpha(s1)- and alpha(s2)-CN in goat's milk. Several analytical parameters were evaluated showing the reliability of this CE method. Coefficients of determination (R2) greater than 99% were obtained and determination limits of 1.23 and 0.98 mg/ml were achieved for alpha(s1)- and alpha(s2)-CN, respectively. The analytical parameters studied in terms of accuracy, precision and recovery were within acceptable limits. Among 18 samples of 4 different genotypes (BB, EE, BF and FF) for alpha(s1)-CN were analysed, different amounts were obtained from the genotypes. PMID- 15553155 TI - Characterization of denatured metallothioneins by reversed phase coupled with on line chemical vapour generation and atomic fluorescence spectrometric detection. AB - A new analytical hyphenated technique is proposed for determination and characterization of thiolic proteins, based on reverse phase chromatography (RPC) coupled on-line with cold vapour generation atomic fluorescence spectrometry (CVGAFS). Proteins are pre-column simultaneously denatured and derivatized in phosphate buffer solution containing 8.0 mol l(-1) urea and p hydroxymercurybenzoate (PHMB). The derivatized proteins are separated on a C4 Vydac Reverse Phase column. Post-column on-line reaction of derivatized denatured proteins with bromine, generated in situ by KBr/KBrO3 in HCl medium, allowed the fast conversion of both the uncomplexed PHMB and of the PHMB bound to proteins to inorganic mercury, also in the presence of methanol in the RPC eluent phase. Hg(II) is selectively detected by AFS in a Ar/H2 miniaturized flame after sodium borohydride reduction to Hg degrees. Under optimized conditions, on-line bromine treatment gives a 98+/-2% recovery of both free and protein-complexed PHMB. The effect of methanol on the sensitivity of Hg(II) detection was studied and controlled. RPC-CVGAFS system has been applied to the analysis of metallothioneins from rabbit liver (MT(RL)) standard solutions, and their commercial isoforms MT-1 and MT-2. The analysis of denatured, PHMB-complexed MTs allowed the determination of the number of thiolic groups complexed by PHMB. It was found that MTs from rabbit liver have 10.0+/-0.3 (MT-1) and 6.7+/-0.3 (MT-2 and MT(RL)) -SH groups complexed by PHMB. The detection limit (LODc) for PHMB in 95% methanol in the optimized conditions was about 9.3 x 10(-9) mol l(-1) and for the denatured MTs LODc was about 8.6 x 10(-10) mol l(-1), taking into account an approximate complexating ratio PHMB:MTs of 7:1. PMID- 15553156 TI - Characterization of winemaking yeast by cell number-size distribution analysis through flow field-flow fractionation with multi-wavelength turbidimetric detection. AB - Yeasts are widely used in several areas of food industry, e.g. baking, beer brewing, and wine production. Interest in new analytical methods for quality control and characterization of yeast cells is thus increasing. The biophysical properties of yeast cells, among which cell size, are related to yeast cell capabilities to produce primary and secondary metabolites during the fermentation process. Biophysical properties of winemaking yeast strains can be screened by field-flow fractionation (FFF). In this work we present the use of flow FFF (FlFFF) with turbidimetric multi-wavelength detection for the number-size distribution analysis of different commercial winemaking yeast varieties. The use of a diode-array detector allows to apply to dispersed samples like yeast cells the recently developed method for number-size (or mass-size) analysis in flow assisted separation techniques. Results for six commercial winemaking yeast strains are compared with data obtained by a standard method for cell sizing (Coulter counter). The method here proposed gives, at short analysis time, accurate information on the number of cells of a given size, and information on the total number of cells. PMID- 15553157 TI - Chromatographic speciation of anionic and neutral selenium compounds in Se accumulating Brassica juncea (Indian mustard) and in selenized yeast. AB - Selenium-accumulating plants such as Brassica juncea (Indian mustard) concentrate the element in plant shoots and roots. Such behavior may provide a cost-effective technology to clean up contaminated soils and waters that pose major environmental and human health problems (phytoremediation). Such ability to transform selenium into bioactive compounds has important implications for human nutrition and health. Element selective characterization of B. juncea grown in the presence of inorganic selenium under hydroponic conditions provides valuable information to better understand selenium metabolism in plants. The present work determines both previously observed organoselenium species such as selenomethionine and Se-methylselenocysteine and for the first time detects the newly characterized S-(methylseleno)cysteine in plant shoots and roots when grown in the presence of selenate or selenite as the only selenium source. A key feature of this study is the complementary role of selenium and sulfur specific chromatographic detection by HPLC with interfaced inductively coupled plasma mass spectrometry (ICP-MS) detection and by derivatization GC with interfaced atomic spectral emission. HPLC-ICP-MS limits of detection for such species were in the range 5-50 ng Se mL(-1) in the injected extracts. Speciation profiles are compared with those of selenium-enriched yeast by both HPLC-ICP-MS and GC-AED. PMID- 15553158 TI - Investigation of the elemental composition and chemical association of several elements in fulvic acids dietary supplements by size-exclusion chromatography UV inductively coupled plasma mass spectrometric. AB - Four fulvic acid dietary supplement samples were obtained for this study with the intention of investigating the elemental composition and association of fulvic acids found in fulvic acid supplements. This was achieved by coupling size exclusion chromatography (SEC) sequentially with UV-vis and inductively coupled plasma mass spectrometric (ICP-MS) detectors. The combination of UV and ICP-MS offered highly sensitive and selective detection. This technique was used in the present study to initially investigate the chemical association of several different elements including, Cr, Co, Ca, Fe, I, Mg, Zn, Se, Cu, Mn, Mo, As, Hg, Pb, and Ag, by observing the elution profile of the fulvic acids obtained with UV detection and matching their retention times with the peaks measured with ICP-MS. The results found based on this type of analysis suggest that there was some association of the elements to the fulvic acids. It was also of interest to observe the stability of these complexes upon human digestion; therefore a gastric digestion was mimicked. In the fulvic acid dietary supplement samples studied, fulvic acids were present in the samples and there was elemental association based on the retention time overlap in the UV as well as the ICP-MS. The fulvic complexes found in the samples were of a low molecular weight As a result of the digestion the SEC-ICP-MS chromatographic profile in some of the samples changed, which may infer that the elemental association had changed. PMID- 15553159 TI - Human exposure to polybrominated diphenyl ethers through the diet. AB - Polybrominated diphenyl ethers (PBDEs), a class of brominated flame retardants, are used in a variety of consumer products being produced in notable quantities. PBDEs have been detected in environmental samples. In recent years, a marked increase in the levels of PBDEs in human biological tissues and fluids, especially breast milk, has been observed in some countries. As for other persistent organic pollutants (POPs), dietary intake is very probably the main route of exposure to PBDEs for the general population. This paper reviews the state of the science regarding human exposure to PBDEs through the diet. Because of the scarce information about it, it is concluded that studies focused on determining PBDE exposure for the population of a number of countries are clearly required. The correlation of PBDE body burdens and dietary intake of PBDEs are also necessary. PMID- 15553160 TI - Polychlorinated naphthalenes in animal aquatic species and human exposure through the diet: a review. AB - Polychlorinated naphthalenes (PCNs) are a group of environmental pollutants, which contain one to eight chlorine atoms per naphthalene molecule, forming a total of 75 possible congeners. Several of the PCN congeners display toxicity similar to that of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) through AhR mediated mechanisms. There are toxicological similarities between PCNs and other well known environmental contaminants such as polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). However, in contrast to these compounds, information on exposure to PCNs for non-occupationally exposed populations is rather scarce. In this article, information on human exposure to PCNs through dietary intake is reviewed. Because this information is very limited and taking into account that most data on PCN levels in potential foods concern to aquatic species, these data are also reviewed. It is concluded that further investigations on dietary intake and potential human health effects of PCNs are clearly necessary. PMID- 15553161 TI - Ruggedness and other performance characteristics of low-pressure gas chromatography-mass spectrometry for the fast analysis of multiple pesticide residues in food crops. AB - Low-pressure gas chromatography-mass spectrometry (LP-GC-MS) using a quadrupole MS instrument was further optimized and evaluated for the fast analysis of multiple pesticide residues in food crops. Performance of two different LP-GC-MS column configurations was compared in various experiments, including ruggedness tests with repeated injections of pesticides in matrix extracts. The tested column configurations employed the same 3 m x 0.15 mm i.d. restriction capillary at the inlet end, but different analytical columns attached to the vacuum: (A) a 10 m x 0.53 mm i.d., 1 microm film thickness RTX-5 Sil MS column; and (B) a 10 m x 0.25 mm i.d., 0.25 microm film thickness DB-5MS column. Under the optimized conditions (compromise between speed and sensitivity), the narrower analytical column with a thinner film provided slightly (<1.1-fold) faster analysis of <5.5 min separation times and somewhat greater separation efficiency. However, lower detection limits for most of the tested pesticides in real extracts were achieved using the mega-bore configuration, which also provided significantly greater ruggedness of the analysis (long-term repeatability of analyte peak intensities, shapes, and retention times). Additionally, the effect of the increasing injection volume (1-5 microl) on analyte signal-to-noise ratios was evaluated. For the majority of the tested analyte-matrix combinations, the increase in sensitivity caused by a larger injection did not translate in the same gain in analyte detectability. Considering the costs and benefits, the injection volume of 2-3 microl was optimal for detectability of the majority of 57 selected pesticides in apple, carrot, lettuce, and wheat extracts. PMID- 15553162 TI - Simple and rapid assay for analyzing residues of carbamate insecticides in bovine milk: hot water extraction followed by liquid chromatography-mass spectrometry. AB - A simple, specific and rapid procedure for determining six largely used carbamate insecticides in bovine whole milk is here presented. This method is based on the matrix solid-phase dispersion technique with heated water as extractant followed by liquid chromatography (LC)-mass spectrometry (MS) equipped with a single quadrupole and an electrospray ion source. Target compounds were extracted from milk by water heated at 90 degrees C. After acidification and filtration, 0.2 mL of the aqueous extract was injected in the LC column. MS data acquisition was performed in the selected ion-monitoring mode, selecting three ions for each target compound. Heated water appeared to be an excellent extractant, since absolute recovery data ranged between 76 and 104% with R.S.D. not larger than 8%. Using butocarboxim (an obsolete carbamate insecticide) as surrogate internal standard, the accuracy of the analysis at three spike levels varied between 85 and 105% with R.S.D. not larger than 9%. On the basis of a signal-to-noise ratio of 10, limits of quantification were estimated to range between 3 ppb (propoxur) and 8 ppb (pirimicarb). The effects of temperature, volume and flow rate of the extractant on the analyte recovery were studied. PMID- 15553163 TI - Cold-temperature stability of five beta-lactam antibiotics in bovine milk and milk extracts prepared for liquid chromatography-electrospray ionization tandem mass spectrometry analysis. AB - The stability of five major beta-lactam antibiotics (amoxicillin, ampicillin, cloxacillin, oxacillin, and penicillin G) in fortified milk and in milk extracts prepared for LC-ESIMS/MS analysis was studied at varying cold temperatures (4, 20, and -76 degrees C). Storage of milk samples at 4 degrees C resulted in measurable losses of all beta-lactams after 6 days (>50% in most cases). Slow degradation of penicillin G, cloxacillin, and oxacillin was observed in milk stored at -20 degrees C, but no losses were recorded at -76 degrees C over 4 weeks. All antibiotics showed good stability at all temperature tested in milk extracts prepared for LC-ESIMS/MS analysis. The results of this study emphasize adherence to adequate sample handling and storage protocols as to reflect residue levels at the time of sample submission. PMID- 15553164 TI - Quantitative determination of chloramphenicol in milk powders by isotope dilution liquid chromatography coupled to tandem mass spectrometry. AB - A method is described for the determination of residues of the illegal antibiotic chloramphenicol (CAP) in milk powders. The analyte is quantified by liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (LC ESI-MS-MS) operating in negative ion multiple reaction monitoring mode (MRM) after a liquid-liquid extraction followed by a clean-up step on solid phase extraction (SPE) cartridge. Because of the presence of two chlorine atoms in the CAP molecule, four specific transition reactions of CAP were monitored by MS-MS in selecting m/z 321 --> 257, 321 --> 152 (35Cl2) and m/z 323 --> 257, 323 --> 152 (37Cl35Cl). Two calibration curves were constructed by plotting the area ratio of m/z 321 --> 152 versus 326 --> 157 and m/z 321 --> 257 versus 326 --> 262 against their corresponding amount ratio. Indeed, even if m/z 321 --> 152 was found to give a higher MS-MS response (calibration curve used by default), an interfering chemical substance was sometimes observed for some milk extracts and not for the transition m/z 321 --> 257. The quantitation method was validated according to the European Union (EU) criteria for the analysis of veterinary drug residues at 0.1, 0.2 and 0.5 microg/kg concentration levels using d5-CAP as internal standard. The decision limit (CCalpha) and detection capability (CCbeta) of CAP in milk were calculated for m/z 321 --> 152 at 0.02 microg/kg and 0.03 microg/kg, respectively, and for m/z 321 --> 257 at 0.02 microg/kg and 0.04 microg/kg, respectively. At the lowest fortification level (i.e. 0.1 microg/kg), repeatability and within-laboratory reproducibility were calculated for m/z 321 - > 257 both at 0.02 microg/kg and for m/z 321 --> 152 at 0.03 and 0.05 microg/kg, respectively. Moreover, the measurement of uncertainty of the analytical method was calculated at the same spiking levels and falls within the precision values of the within-laboratory reproducibility. This method can be applied to several types of milk powders (e.g. full cream, skim) and can serve as a monitoring tool to avoid that unacceptable levels of residues of CAP enter the food chain. PMID- 15553165 TI - Multiresidue method for the detection of tranquillisers, xylazine, and a beta blocker in animal production by liquid chromatography-tandem mass spectrometry. AB - Tranquillisers are often used in animal production, especially in pigs to calm them before transport to the slaughterhouse. The use of certain substances (derived from phenothiazine) is totally prohibited, whilst other compounds (butyrophenone and beta-blockers) are regulated through the establishment of MRLs. A physico-chemical detection method based on liquid chromatography-tandem mass spectrometry is described. Validation was carried out according to the criteria laid down in Directive 2002/657/EC. The method was also used to detect and quantify these substances in treated animals. PMID- 15553166 TI - Oxidative degradation of food dye E133 Brilliant Blue FCF Liquid chromatography electrospray mass spectrometry identification of the degradation pathway. AB - This paper is devoted to the evaluation of the degradation pathway of the E133 Brilliant Blue FCF (C.I. 42090) that is largely used in the food industry. The degradation is studied in oxidation conditions obtained by addition of potassium persulfate at different persulfate to dye molar ratios under natural sunlight irradiation. The degradation pathway of the dye passes through a species coloured in dark blue and then gives rise to uncoloured species. Due to the low volatility and the poor thermal stability of the dye, reversed-phase liquid chromatography associated to mass spectrometry and tandom mass spectrometry was employed to follow the kinetics of degradation and identify some intermediates. The identification of organic species still present in the decoloured dye and the value of COD obtained in these conditions show evidence that complete decolorization does not correspond to complete mineralisation. No direct information of toxicity is available for the uncoloured degradation products but the further formation of aromatic amines can not be excluded. PMID- 15553167 TI - Simultaneous detection of type A and type B trichothecenes in cereals by liquid chromatography-electrospray ionization mass spectrometry using NaCl as cationization agent. AB - A LC/MS method for the simultaneous determination of both type A and type B trichothecenes by using an electrospray ionization (ESI) interface in the positive ionization mode with a single quadrupole analyzer is described. In order to enhance the ionization of both groups of trichothecenes, the sodium ion was used as cationization agent by adding sodium chloride to the eluent. All LC/MS parameters were optimized. The newly developed LC/ESI-MS method was applied to the analysis of a wheat reference material and cereal-based foods and feeds. PMID- 15553168 TI - Concentration of ochratoxin A in wines from supermarkets and stores of Valencian Community (Spain). AB - Ochratoxin A (OTA) is a mycotoxin produced by fungi species belonging to the genera Aspergillus and Penicillium being isolated in alcoholic beverages. The aim of this work is developed and applied a procedure for the analysis of OTA in wines. An analytical method based on immunoaffinity column (IAC) for clean-up, liquid chromatography with fluorescence detection (LC-FD), and LC-FD after of OTA methylation was used to determine the occurrence of OTA in wines. Recoveries of this mycotoxin spiked to red wines at 0.5 ng/ml level were >90% with an average of relative standards deviations of 4%. Furthermore, 116 wine samples from designation of origin (DO) and three samples from food stores of Valencian Community (Spain) were examined for the occurrence of OTA being the levels of this mycotoxin ranged from <0.01 to 0.76 ng/ml. Finally, the estimated daily intake of OTA in this study was 0.15 ng/kg bw per day. PMID- 15553169 TI - Determination of volatile N-nitrosamines in irradiated fermented sausage by gas chromatography coupled to a thermal energy analyzer. AB - Volatile N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR) in irradiated pepperoni and salami sausages were determined using a gas chromatography coupled to a thermal energy analyzer (GC-TEA). These fermented sausages with aerobic or vacuum packaging were irradiated at 0, 5, 10, and 20 kGy, and then stored for 4 weeks at 4 degrees C. Both NDMA and NPYR in the fermented sausage were significantly reduced by irradiation. The vacuum packaging showed significantly lower (P < 0.05) N-nitrosamine levels than that of the aerobic ones. After storage, the contents of NDMA and NPYR in the irradiated sausage were lower than those of the non-irradiated control. Results indicated that a high dose of irradiation (>10 kGy) was needed to reduce the carcinogenic N nitrosamines in the fermented sausage during storage and the GC-TEA analysis was effective in determining the N-nitrosamines in irradiated meats even at low trace levels. PMID- 15553170 TI - Determination of heterocyclic amines by liquid chromatography-quadrupole time-of flight mass spectrometry. AB - This paper discusses the applicability of coupling liquid chromatography (LC) to mass spectrometry (MS) using a time-of-flight (TOF) mass analyzer for the analysis of heterocyclic amines (HAs). Accurate mass measurement (<2 mDa) with both MS-MS and in-source CID MS-MS was carried out to confirm the elemental composition of some fragments previously reported. Some isobaric assignments (fragments containing N versus CH2 and NH3 versus CD2H) were distinguished by taking advantage of the resolution provided by the TOF mass analyzer. On the other hand, the LC-MS analysis of HAs in MS acquisition mode was also performed. Quality parameters of the method were established. The linearity range extended over three orders of magnitude, limits of detection were in the pg level and good short-term precision values (R.S.D., 1.2-8.0%) were obtained. The LC-ESI-TOF method was applied to the determination of HAs in a lyophilized meat extract and the results obtained were comparable to those given by MS-MS with triple quadrupole and ion trap instruments. PMID- 15553171 TI - Immunomodulation of human natural killer cell cytotoxic function by organochlorine pesticides. AB - Organochlorine pesticides are used worldwide. To our knowledge there have been no studies dealing with the effects of these agents under in vitro conditions on human natural killer (NK) cell cytotoxic function. NK cells play a central role in immune defense against tumor development and viral infections. Thus, any agent that interferes with the ability of NK cells to lyse their targets could increase the risk of tumor incidence and/or viral infections. In this study, we examined the effects of organochlorine pesticides and some of their breakdown products on the ability of human NK cells to lyse tumor cells. A total of 11 compounds were tested. The compounds were tested in both purified NK cells as well as a cell preparation that contained other mononuclear cells (predominantly T cells) and NK lymphocytes (referred to as T/NK cells). Lymphocytes were exposed to the compounds for periods of time ranging from 1 hour to 6 days. Exposure of highly purified NK cells to 5 microM alpha-chlordane, gamma-chlordane, 4,4'-DDT, heptachlor, oxychlordane, or pentachlorophenol (PCP) inhibited their ability to destroy K562 tumor-cells by 88+/-5, 92+/-8, 61+/-13%, 64+/-10%, 69+/-11%, 76+/ 12%, respectively, after a 24 h exposure. The loss of cytotoxic function seen with alpha-and gamma-chlordane remained essentially constant out to 6 days, while that seen with 4,4'-DDT, oxychordane and PCP increased with longer exposures (6 d). PCP was the most effective of the compounds tested at decreasing NK function. Of the compounds that caused decreased lytic function when tested in purified NK cells, only PCP and oxychordane decreased the lytic function of the T/NK cell preparation after any exposure. The results provide evidence of relative toxic potential for the 11 compounds and their immunomodulatory effects on other mononuclear cells (such as T-cells, B-cells, and monocytes) as well as NK lymphocyte function. PMID- 15553172 TI - Outcomes following abuse of methanol-containing carburetor cleaners. AB - INTRODUCTION: Carbureter cleaners may contain methanol and are abused via inhalation. Toxicity resulting from the methanol component of these products is poorly described. METHODS: We conducted a retrospective poison center chart review over a four-year period (3/98-3/02) of outcomes following methanol containing carbureter cleaners (MCC) exposure. Inclusion criteria were: (1) use of MCC, (2) evaluation in health care facility (HCF), (3) no known co-ingestion exposure and (4) at least 12 hour follow-up. RESULTS: 33 cases were reviewed with 11 cases excluded because of significant co-ingestions. Of the remaining 22 cases the mean age was 17 [range: 14-41] years old with 90% of cases between 14 and 17 years old. Six women and 16 men were in the study. Six of 22 cases had acidosis (serum bicarbonate < or =22 mmol/L or pH < or =7.35), 100% of patients had neurological symptoms (ataxia, etc.) and 14/22 had vomiting on presentation. Three patients received treatment with ethanol (1) and fomepizole (2). All others received intravenous fluids (15) or no treatment (4). Mean serum methanol concentration was 28mg/dl [range: 0-341 with 17/22 developing acidosis. Serum methanol was obtained at a mean of 3.5 hours [range 1-7 hours] post use. All metabolic disturbances resolved within 24 hours except in one patient (41 years old) in which her disturbances resolved within 72 hours. No patient developed visual disturbances or neurological sequealae. CONCLUSIONS: Significant toxicity following inhalation of MCC was rare with symptoms improving without aggressive care (dialysis, alcohol dehydrogenase blockade). PMID- 15553173 TI - Changes in hematological, hemostatic and biochemical parameters induced experimentally in rabbits by Loxosceles gaucho spider venom. AB - Human accidents caused by Loxosceles spiders may result in local dermal necrosis and, in some cases, severe systemic reactions - such as intravascular hemolysis, disseminated intravascular coagulation (DIC), renal failure and death. Since many aspects of envenomation by Loxosceles spiders remain unclear, we studied the hematological and hemostatic responses induced by the i.d. injection of 10 microg/kg Loxosceles gaucho venom in rabbits. For this purpose, total blood cell count, platelet function, coagulation tests and biochemical parameters were analysed at 3, 24, 48, 72 and 120 hours after venom administration. Thrombocytopenia and leukopenia were noted at 3 and 24 hours. Histopathological analysis of the skin lesion, performed at 24 hours after venom administration, showed a massive presence of leukocytes and platelets, hemorrhage and thrombus formation at the injection site. At 72 and 120 hours, neutrophilic leukocytosis and thrombocytosis were observed. Platelet hyperaggregation was noticeable at 48 and 72 hours. Haptoglobin and fibrinogen levels were elevated early and remained in high levels over time. Significant increases in coagulation factors V, VII, VIII, IX, X and XI were noted at 120 hours. The results showed that neither intravascular hemolysis nor DIC occurred. However, the early onset of thrombocytopenia and leukopenia are important findings that may be related to dermal necrosis formation during loxoscelism. PMID- 15553174 TI - Cytotoxicity of the phytosterol diosgenin and its derivatives in rat cultured hepatocytes and V79 fibroblasts. AB - In this work, the cytotoxic effects of some spirostane derivatives were examined in cultured hepatocytes and V79 fibroblasts using different viability assays. The derivatives were obtained by modifying the A and B rings of diosgenin. Diosgenin and its derivatives were more toxic in V79 fibroblasts (IC50 40-300 microM) than in hepatocytes (IC50 280-1000 microM). Inhibition of cytochrome P450IIIA in cultured hepatocytes by incubation with 1 mM cimetidine did not alter the toxicity of these compounds in these cells. These observations suggest that other pathways of detoxification may be involved in hepatocytes. In conclusion, the compounds studied merit investigation for their potential pharmacological and industrial applicability. PMID- 15553175 TI - Ochratoxin A levels in human plasma and foods in Lebanon. AB - Ochratoxin A (OTA) is a widespread mycotoxin which contaminates food such as cereals, beer, coffee, wine and products of animal origin. OTA is known for its nephrotoxic, immunotoxic and carcinogenic properties. The prevalence of OTA in human blood and foodstuffs has been investigated in many countries. In this study, exposure of the Lebanese population to OTA was evaluated and the contamination of the most commonly consumed foods in Lebanon by OTA was assessed. Plasma samples from healthy individuals and also cereals and beer samples obtained from markets were collected from the different regions of Lebanon. OTA was detectable in 33% of tested plasma samples (n =250) with a concentration ranging from 0.1 to 0.87 ng/mL and a mean of 0.17+/-0.01ng/mL. No sex and age differences were found. The frequency of OTA-positive plasma samples obtained in the South of Lebanon and in the Bekaa valley (50 and 47%, respectively) was significantly higher compared to plasma samples obtained in the Beirut/Mount Lebanon region (19%). Food analyses showed that wheat, burghul and beer were contaminated with a mean value of 0.15+/-0.03 microg/kg, 0.21+/-0.04 microg/kg and 0.19+/-0.12 ng/mL, respectively. These data suggest that the Lebanese population is exposed to OTA through food ingestion at concentrations lower than the tolerable daily intake. PMID- 15553176 TI - Difficulties in assessing brain death in a case of benzodiazepine poisoning with persistent cerebral blood flow. AB - Assessing brain death may sometimes be difficult, with isoelectric EEG following psychotrope overdoses or normal cerebral blood flow (CBF) persisting despite brain death in the case of ventricular drainage or craniotomy. A 42-year-old man, resuscitated after cardiac arrest following a suicidal ingestion of ethanol, bromazepam and zopiclone, was admitted in deep coma. On day 4, his brainstem reflexes and EEG activity disappeared. On day 5, his serum bromazepam concentration was 817 ng/ml (therapeutic: 80-150). The patient was unresponsive to 1 mg of flumazenil. MRI showed diffuse cerebral swelling. CBF assessed by angiography and Doppler remained normal and EEG isoelectric until he died on day 8 with multiorgan failure. There was a discrepancy between the clinically and EEG assessed brain death, and CBF persistence. We hypothesized that brain death, resulting from diffuse anoxic injury, may lead, in the absence of major intracranial hypertension, to angiographic misdiagnoses. Therefore, EEG remains useful to assess diagnosis in such unusual cases. PMID- 15553177 TI - How the government broke its Bristol inquiry pledge. PMID- 15553178 TI - What are the odds? Can the Confed straddle two horses in its expanded role as employers' organisation? PMID- 15553179 TI - Evidence ideas. The human touch. PMID- 15553180 TI - Data briefing. Public opinion. PMID- 15553181 TI - Clinical management where medicine meets management. Knowledge is power. AB - --Improving knowledge and understanding of diabetes among primary care professionals and patients has triggered service improvements in one PCT. - Diabetes care once given in hospital is now provided in primary care. --Referrals to hospital have halved and waiting time reduces. PMID- 15553182 TI - Elderly people's services. Home truths. AB - --A study assessed whether more older people would avoid institutional care if current home care technologies were utilised fully. --A number of people were found to benefit from different technology bundles and at a lower cost than institutional care. --For this approach to succeed, high application of comprehensive geriatric assessment would be needed. PMID- 15553183 TI - Mental health. Cents and sensibility. PMID- 15553184 TI - HSJ people: regional focus on the midlands. Central spark. PMID- 15553185 TI - Buildings: from the drawing board. PMID- 15553186 TI - Buildings: race against time. PMID- 15553187 TI - Buildings: estates of disarray. PMID- 15553188 TI - Buildings: design for life. PMID- 15553189 TI - Vaccination: a victim of its own success? PMID- 15553190 TI - What is your diagnosis? Juvenile-onset generalised demodicosis. PMID- 15553191 TI - Cytological features of canine ovarian tumours: a retrospective study of 19 cases. AB - The cytological features of 19 histologically confirmed canine ovarian tumours were retrospectively examined. Seven cases were cytologically classified as papillary adenocarcinoma, eight cases as granulosa cell tumours, two cases as mature ovarian teratomas, one case as a dysgerminoma and one case as a mixed granulosa cell tumour/dysgerminoma. On cytology, papillary adenocarcinoma was characterised by a papillary glandular pattern and tight cohesiveness. Granulosa cell tumours showed monolayered clusters of loosely cohesive granulosa cells. Call-Exner-like bodies were found in five of seven cases. Granulosa cells appeared to be heterogeneous and usually contained several intracytoplasmic vacuoles. Teratoma was characterised cytologically by keratin debris (two cases) and a mixture of epithelial cells with sebaceous, basaloid, columnar/palisading or ciliated appearance (one case). The dysgerminoma contained severely atypical round cells admixed with small lymphocytes. The mixed dysgerminoma/granulosa cell tumour had a mixture of germinal and granulosa cells. Cytological diagnosis was in agreement with histopathology in 18 of 19 (94.7 per cent) cases. PMID- 15553192 TI - A retrospective study of clinical findings, treatment and outcome in 143 dogs with pericardial effusion. AB - A retrospective study of 143 dogs with pericardial effusion is presented, including a statistical analysis of survival time. Cases were classified into those in which a mass was seen on echocardiography (echo-positive) and those in which no mass could be identified (echo-negative). Forty-four dogs were echo positive and 99 were echo-negative. The median survival time (MST) was 1068 days for echo-negative dogs and 26 days for echo-positive dogs. Dogs with a history of collapse were more likely to present with a mass on echocardiography. Those presenting with collapse had an MST of 30 days compared with 605 days for those without collapse. Echo-negative dogs tended to present with ascites and generally had a larger volume of pericardial effusion. The median survival for dogs presenting with ascites was 605 days compared with 45 days for those without ascites. Among echo-negative dogs, 64 per cent had a relapse of their effusion. Subtotal pericardiectomy was performed in 31 echo-negative dogs. The procedure had a perioperative mortality of 13 per cent but provided a favourable long-term prognosis. Dogs undergoing pericardiectomy had a median survival of 1218 days compared with 532 days for those not undergoing surgery. PMID- 15553193 TI - Acute B cell lymphoblastic leukaemia in a 12-week-old greyhound. AB - A 12-week-old greyhound had a two-day history of lethargy, inappetence and shifting lameness. Clinical examination revealed pyrexia and hepatosplenomegaly. Haematological examination showed anaemia, thrombocytopenla, neutropenla and large numbers of atypical mononuclear leucocytes. A diagnosis of acute B cell lymphoblastic leukaemia was made following flow cytometric immunophenotyping of the leucocytes. The owner declined further evaluation and the dog was treated symptomatically with antibiotics. After a brief improvement, the dog's condition deteriorated and it was euthanased four days after initial presentation. The case was unusual because acute lymphoid leukaemia in the dog is most frequently reported in mature animals. This is in contrast to humans, where acute leukaemia is one of the most common childhood cancers. PMID- 15553194 TI - Two cases of pancreatic neoplasia in British ferrets (Mustela putorius furo). AB - Two six-year-old male neutered polecat ferrets (Mustela putorius furo) were presented for the investigation of acute collapse or periodic weakness and weight loss. While blood biochemistry revealed hypoglycaemia in both cases, diagnosis of an insulin-secreting neoplasia was confirmed by exploratory surgery in one case and supported by the use of an insulin assay in the other. Subsequent histopathological examination showed the former to be a pancreatic islet cell carcinoma and the latter to be a pancreatic islet cell adenoma. While neoplasia of the pancreas commonly affects ferrets in the USA, there appears to be only one previous report from the UK. PMID- 15553195 TI - Chronic renal failure in an English bull terrier with polycystic kidney disease. AB - An entire female English bull terrier, aged five years and one month, was diagnosed with polycystic kidney disease by renal ultrasonography. It had thickening and abnormal motion of the mitral valve on 2D and M mode echocardiography, and left ventricular outflow tract obstruction, characterised by turbulence in the left ventricular outflow tract and elevated aortic blood flow velocity, detected by colour flow and spectral Doppler echocardiography, respectively. Two years later, haematology, serum biochemistry and urinalysis data suggested the presence of compensated renal failure. The dog was euthanased at 10 years and eight months of age, with haematology, serum biochemistry and urinalysis data Indicating decompensated chronic renal failure. Postmortem examination confirmed polycystic kidney disease, chronic renal disease, mitral and aortic valvular myxomatous degeneration, and mixed mammary neoplasia. This case demonstrates that bull terriers with polycystic kidney disease may develop associated chronic renal failure. PMID- 15553196 TI - Juvenile nephropathy in two related Pembroke Welsh corgi puppies. AB - Juvenile nephropathy has been documented in many breeds. Two related Pembroke Welsh corgi puppies presented at three and five months of age, respectively, for evaluation of lethargy, diarrhoea, poor body condition, polyuria and proteinuria. Based upon the clinical presentation, urinalysis and serum biochemistry, chronic renal failure was diagnosed. Renal histopathology was consistent with juvenile nephropathy, revealing lesions similar to the juvenile renal disease of dobermann dogs. To the authors' knowledge, this is the first report of juvenile nephropathy in related Pembroke Welsh corgi dogs. Familial nephropathy should now be considered as a differential diagnosis in cases of young Pembroke Welsh corgi dogs presenting with clinical signs indicating renal failure. PMID- 15553197 TI - A nasal dermoid sinus in an English bull terrier. AB - A five-year-old, entire female English bull terrier was presented with a six-week history of intermittent facial swelling and nasal pain, following an incident of nasal trauma. A small opening was present at the mucocutaneous junction on the dorsal nasal planum. Examination under general anaesthesia allowed catheterisation of this opening and confirmed the presence of a tract passing caudally. Plain radiographic examination of the region was unremarkable. Positive contrast sinography demonstrated contrast material filling a midline tract that passed caudally and subcutaneously towards the nasal bone. This tract was surgically excised. Histopathological examination of the excised tissue, together with the clinical findings, led to the diagnosis of nasal dermoid sinus. PMID- 15553198 TI - Proposed changes to medicines legislation. PMID- 15553199 TI - Who should regulate veterinary nurses? PMID- 15553200 TI - When jobs go pond hopping: outsourcing has become a major economic issue, and healthcare delivery may next fall victim. PMID- 15553201 TI - Casting a wide net: foreign-born workers are essential to the healthcare industry -if you can jump through the hoops to recruit them. PMID- 15553202 TI - The brat pack: new poll concludes that ill-mannered physicians are commonplace. PMID- 15553203 TI - The first helicopter out: with choices narrowing, Americans will inevitably go overseas for care. PMID- 15553204 TI - Asthma and aspergillus. PMID- 15553205 TI - Antituberculosis drug resistance and associated risk factors in the European section of Turkey. AB - BACKGROUND: We carried out this study in order to establish the prevalence of antituberculosis drug resistance in Mycobacterium tuberculosis strains and to determine risk factors for the development of resistance in Trakya region of Turkey. METHODS: Pattern of drug resistance in 214 M. tuberculosis isolates from patients with tuberculosis treated at the regional tuberculosis dispensaries were included in the study. RESULTS: Isolates of 105 (49.1%) were resistant to only one drug, and 62 (29.0%) were resistant to more than one drug. The total resistance rates to streptomycin, isoniazid, rifampicin, ethambutol and isoniazid + rifampicin were 29.0%, 27.1%, 21.5%, 10.3% and 11.6%, respectively. The secondary resistance rates in all drugs and combinations were higher than primary resistance rates (p<0.001). Step wise logistic regression revealed that (i) non compliance with treatment increases the chances of development of resistance by 15 times [p<0.00001, 95% confidence intervals (95% CI) : 4.16 to 56.70], and (ii) a regimen of inadequate treatment increases the chance of development of drug resistance by 10.5 times (p<0.01, 95% CI=2.66 to 49.80). CONCLUSIONS: We propose that specially trained physicians should institute antituberculosis therapy and medication should be practiced under direct observation in this region. PMID- 15553206 TI - Morphological variations of the lung fissures and lobes. AB - BACKGROUND: Knowledge of the position of fissures of the lungs is necessary for the appreciation of lobar anatomy and thus locating the bronchopulmonary segments. METHODS: Morphological variations of fissures and lobes of the lungs were studied in 30 pairs of lungs from cadavers. RESULTS: Five right-sided lungs showed absence of horizontal fissure, 19 showed incomplete horizontal fissure. Eleven right-sided and 14 left-sided lungs showed incomplete oblique fissure and two right-sided lungs showed both absence of horizontal fissure and an incomplete oblique fissure. Accessory fissure was seen in three left-sided and one right sided lungs. CONCLUSIONS: Awareness regarding anatomical variations is essential for performing lobectomies and segmental resection and interpreting radiological images. PMID- 15553207 TI - Role of simple needle aspiration in the management of pneumothorax. AB - OBJECTIVES: To determine the role of simple needle aspiration in the management of pneumothorax. DESIGN: All patients presenting with a pneumothorax requiring intervention were included in this prospective study. Patients who were very sick or had tension pneumothorax were treated with direct intercostal chest tube drainage (ICTD) and others were subjected to simple aspiration. The procedure was deemed successful, if after aspiration the lung expanded completely or symptoms were relieved with residual pneumothorax of less than 15% of hemithorax. In case of failed aspiration ICTD was carried out. RESULTS: Fifty-seven patients with 59 pneumothoraces were included in the study. Of these, 24 were treated with direct ICTD; 35 (12 spontaneous, 11 secondary and 12 iatrogenic pneumothoraces) were subjected to simple aspiration. Ten (83.3%) of the primary, 1 (9.6%) of the secondary and 11 (91.7%) of the iatrogenic pneumothoraces responded to simple aspiration. There were no significant complications. The pain perceived and the duration of hospital stay was significantly lesser in the simple aspiration group. CONCLUSIONS: Simple aspiration should be the initial modality of treatment for primary spontaneous and iatrogenic pneumothoraces. For secondary spontaneous pneumothorax and in conditions where pleurodesis is indicated, ICTD remains the treatment of choice. PMID- 15553208 TI - Non small cell lung cancer (NSCLC): current status and future prospects. AB - The incidence of lung cancer is rising dramatically and it is now the commonest cause of mortality and morbidity not only in the industrialised countries, but in developing nations like India as well. Tobacco smoking has consistently been demonstrated to be an important aetiological factor, though lung cancer occurs in non-smokers also. In spite of great advances in radiological and molecular diagnostic techniques, the ideal screening marker for early detection of lung cancer has still not been found. Histological diagnosis and staging is essential for selecting the mode of therapy in patients with lung cancer. Stages I and II are amenable to surgery. However, advanced stage III and IV need an individualised combination of surgery, chemotherapy and radiotherapy. With a better understanding of the cellular mechanisms operating in carcinogenesis, newer target specific modalities are being developed and tested in order to stem this disease, which threatens to assume epidemic proportions. PMID- 15553209 TI - Lymphangioleiomyomatosis: a rare cause of breathlessness. AB - A 24-year-old female presented with complaints of cough with scanty expectoration, breathlessness on exertion and chest pain for the last three years. These symptoms had appeared during the 12th week of her third pregnancy. She was given anti-tuberculosis treatment at another hospital for nine months without any improvement in symptoms. Four years ago she had been diagnosed to have leprosy of borderline variety for which she had received treatment. On examination, she was tachypnoeic with a respiratory rate of 33 breaths per minute. She had clubbing and small, discrete and firm lymph nodes in the anterior cervical region. Chest examination revealed wheezing with bibasilar end inspiratory crepitations. PMID- 15553210 TI - Pulmonary lymphangioleiomyomatosis: a rare presentation. AB - Pulmonary lymphangioleiomyomatosis (LAM) is a rare idiopathic, cystic disease. We report a case of LAM who presented with nonchylous, bilateral pleural effusion and progressive respiratory failure. PMID- 15553211 TI - Rare case of pulmonary joint, cardiac and mediastinal involvement in rheumatoid arthritis with bronchial asthma and allergic rhinitis. AB - It is extremely uncommon to find a patient with rheumatoid arthritis with pulmonary and cardiac manifestations together with co-existent intrathoracic lymphadenopathy. We report the case of a 40-year-old female with rheumatoid arthritis with rheumatoid lung disease with severe aortic regurgitation, congestive heart failure, bronchial asthma and allergic rhinitis. PMID- 15553212 TI - Mediastinal cavernous haemangioma. AB - A 44-year-old man presented with palpable right supraclavicular swelling having no symptoms pertaining to respiratory tract. A routine chest radiograph showed an anterior mediastinal mass. Computerised tomographic scan (CT-scan) of chest showed an enhancing and homogeneous mass in anterior mediastinal space with few tiny calcific specks within the mass. Biopsy of cervical swelling showed cavernous haemangioma with chronic non-specific lymphadenitis. A sternotomy was undertaken and an encapsulated 9x7.5x5 cm, dark purplish mass was seen in anteior mediastinum adherent to pericardium and right pleura. The cut-surface was elastic and soft. Histology confirmed cavernous haemangioma. Post-operative course was uneventful. Mediastinal cavernous haemangioma is extremely rare and surgical excision is the treatment of choice. PMID- 15553213 TI - Tuberculous abscess of the abdominal wall and multiple splenic abscesses in an immunocompetent patient. AB - We report a case of a 39-year-old human immunodeficiency virus (HIV)--negative male who presented with a progressively increasing swelling in the left hypochondrium. He did not manifest fever or toxaemic symptoms. Computerised tomographic scan (CT scan) of the abdomen revealed an abscess in the anterior wall and multiple splenic abscesses. Fine needle aspiration from the abscesses in the anterior abdominal wall and the spleen confirmed the diagnosis of tuberculosis as the aetiology. The patient responded well to antituberculosis treatment and the abscesses regressed considerably. PMID- 15553214 TI - Pulmonary paragonimiasis. AB - A 27-year-old married male from Lamdeng village of Imphal West District, Manipur presented with cough, fever, haemoptysis, anorexia and weight loss, and was treated with antituberculosis drugs continuously for one year without improvement. He also had history of ingestion of raw crabs. Chest radiograph revealed right midzone lesion with cavitation. Laboratory investigations revealed peripheral blood eosinophilia, elevated erythrocyte sedimentation rate and sputum smear revealed eggs of Paragonimus westermani. He responeded well to treatment with praziquantel 25 mg/kg, three times a day for three days and is doing well on follow-up. Pulmonary paragonimiasis must be considered in the differential diagnosis of slowly resolving pneumonias, especially in the appropriate clinical setting because effective treatment with praziquantel can be rewarding. PMID- 15553215 TI - The value of USP public standards for therapeutic products. PMID- 15553216 TI - "Programmed polymeric devices" for pulsed drug delivery. AB - Pharmaceutical research strives to design drug delivery systems that respond to therapeutic needs. Considering the facts that physiologic parameters (e.g., heart rate, blood pressure, and plasma concentration of hormones, plasma proteins, and enzymes) display constancy over time, drug delivery systems with a constant release profile have been designed. However, because of circadian rhythms in physiologic parameters and pathologic conditions (e.g., asthma, angina pectoris), the conventional paradigm concerning drug concentrations "the flatter the better" may not be what the organism may need. Instead, to correlate with our biological needs, "precisely timed drug delivery," which could be accomplished with "programmable dosage forms," is required. Precisely timed drug delivery may maximize therapeutic efficacy, may minimize dose frequency, and may reduce toxicity by avoiding side effects and drug tolerance. This paper outlines the concepts that have been proposed to release drugs in a pulsed manner from pharmaceutical devices. PMID- 15553217 TI - The arthrotropism of macromolecules in adjuvant-induced arthritis rat model: a preliminary study. AB - PURPOSE: To study the accumulation of macromolecules into the arthritic joints and the possible applications of such phenomenon. METHODS: The accumulation of plasma albumin in the joints of adjuvant-induced arthritis (AIA) rat model was first visualized with Evans blue injection. A N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer contrast agent was then synthesized and injected into the AIA rats to allow qualitative examination of biodistribution and pharmacokinetics of the injected macromolecule with magnetic resonance imaging (MRI). Vital organs and the diseased joints were isolated and examined histologically to correlate with the MRI findings. RESULTS: Deep blue color developed around the arthritic joints of the AIA rat a few hours after the injection of Evans blue. MR imaging of the AIA rats injected with polymer contrast agent demonstrated a gradual but very strong accumulation of the injected polymer in the arthritic joints, which lasted for 1-2 days. Observed differences in the concentration of the injected polymer in the joints correlated with disease severity as assessed histologically. CONCLUSIONS: Profound arthrotropism of macromolecules in the AIA rat model was demonstrated with various imaging tools. These observations should help in the conceptual and practical design of novel macromolecular delivery systems for the imaging and treatment of rheumatoid arthritis. PMID- 15553218 TI - Reactivity of 1,4-dihydropyridines toward SIN-1-derived peroxynitrite. AB - PURPOSE: To study the reactivity of C4-substituted 1,4-dihydropyridines (1,4 DHP), with either secondary or tertiary nitrogen in the dihydropyridine ring, toward SIN-1-derived peroxynitrite in aqueous media at pH 7.4. METHODS: Reactivity was followed by changes in the absorptivity of the UV-Vis bands corresponding to 1,4-DHP. Gas Chromatography/ Mass Spectrometer (GC-MS) and Electron Paramagnetic Resonance (EPR) spin trap techniques were used to characterize the final product and the intermediates of the reaction, respectively. RESULTS: 1,4-DHPs significantly reacted toward peroxynitrite at varied rates, according to the calculated kinetic rate constants. By EPR spectroscopy, a carbon-centered radical from the 1,4-DHP was intercepted with N tert-butylamine-alpha-phenylnitrone (PBN), as the intermediate for the reaction with peroxynitrite. Likewise, the oxidized derivative (i.e., the pyridine) was identified as the final product of the reaction by GC-MS. By using the technique of deuterium kinetic isotope effect, the participation of the hydrogen of the 1 position on the 1,4-DHP ring was shown not to be the rate-limiting step of the reaction. CONCLUSIONS: The direct participation of the 1,4-DHP derivatives in the quenching of SIN-1-derived peroxynitrite has been demonstrated. Kinetic rate constant of tested 1,4-DHP toward peroxynitrite showed a direct relationship with the oxidation peak potential values; that is, compounds reacting faster were more easily oxidized. PMID- 15553219 TI - Miniature device for aqueous and non-aqueous solubility measurements during drug discovery. AB - PURPOSE: A miniature device was developed for the measurement of aqueous and non aqueous equilibrium solubility during drug discovery. The solubility values obtained using the miniature device were compared to those obtained using the conventional shake-flask method. METHODS: The aqueous solubility of six structurally diverse compounds, the solubility of carbamazepine in various cosolvent systems, and the pH-solubility profile of saquinavir were determined using the miniature device. The device contains a multichannel cartridge pump and a Tygon tubing that is mounted on the pump with two ends linked by a syringe filter. The drug slurry was filled into the tubing and circulated inside, continually passing through the syringe filter. At the end of the experiment, the filtrate was collected and analyzed directly by High-Pressure Liquid Chromatography (HPLC). The solubility was also determined by the shake-flask method. RESULTS: The solubility values determined by the miniature device were in good agreement with those measured by the conventional shake-flask method. CONCLUSIONS: The miniature device provides a unique way of testing aqueous and non-aqueous equilibrium solubility in a microscale setting. With approximately 1 mg of compound, it is possible to determine the entire pH-solubility profile. The device is useful for solubility screening during lead optimization and candidate selection in early drug discovery, when compound supply is limited. It can also be used for screening solubility in non-aqueous systems to select vehicles for preclinical in vivo studies. PMID- 15553220 TI - Stability and solubility of celecoxib-PVP amorphous dispersions: a molecular perspective. AB - PURPOSE: The purpose of the current study is to evaluate the solubility advantage offered by celecoxib (CEL) amorphous systems and to characterize and correlate the physical and thermodynamic properties of CEL and its amorphous molecular dispersions containing poly(vinylpyrrolidone) (PVP). METHODS: The measurement of crystalline content, glass transition temperatures, and enthalpy relaxation was performed using differential scanning calorimetry. Solubility and dissolutions studies were conducted at 37 degrees C to elucidate release mechanisms. Further, the amorphous systems were characterized by polarized light microscopy and X-ray powder diffraction studies. RESULTS: The PVP content has a prominent effect on the stability and solubility profiles of amorphous systems. A dispersion of 20% w/w PVP with CEL resulted in a maxima in terms of solubility enhancement and lowering of relaxation enthalpy. The release of drug from amorphous molecular dispersions was found to be drug-dependent and independent of the carrier. CONCLUSIONS: The solubility enhancement and enthalpy relaxation studies with respect to PVP concentration helped in a better prediction of role of carrier and optimization of concentration in the use of solid dispersions or amorphous systems. The drug release mechanism is drug-controlled rather than carrier controlled. PMID- 15553221 TI - Extended nasal residence time of lysostaphin and an anti-staphylococcal monoclonal antibody by delivery in semisolid or polymeric carriers. AB - PURPOSE: Eradication of Staphylococcus aureus nasal colonization reduces the risk of nosocomial and community acquired infections with this organism. This study describes the formulation and use of lysostaphin and BSYX-A110, an anti lipoteichoic acid monoclonal antibody, for eradication of S. aureus nasal colonization. METHODS: Lysostaphin was formulated into a hydrophilic cream that forms an emulsion with the secretions of the nasal mucosa, and aqueous formulations of BSYX-A110 were made containing the mucoadhesive polymers polystyrene sulfonate and chitosan. Intranasal pharmacokinetics of the drugs was measured in mice and cotton rats. RESULTS: Lysostaphin formulated in the cream increased nasal retention of the drug by 10-fold at 3 h post-cream installation and 50-fold at 24 h as compared to lysostaphin in saline drops. Furthermore, the levels of lysostaphin in the nose 24 h post-cream instillation are still above the minimum bactericidal concentration for most bacterial strains. The liquid polymer formulations also resulted in prolonged retention of antibody in the nose, with 4-fold higher levels at 3 h post-instillation as compared to antibody in saline drops. CONCLUSIONS: These results demonstrate that cream and polymer delivery systems significantly decrease the clearance rate of lysostaphin and BSYX-A110 from the nose, thereby enhancing their therapeutic potential for eradicating S. aureus nasal colonization. PMID- 15553222 TI - Effect of gender and device mouthpiece shape on bolus insulin aerosol delivery using the AERx pulmonary delivery system. AB - PURPOSE: A study was designed to compare differences in insulin aerosol deposition profiles in healthy male and female subjects, as well as examine the effect of mouthpiece cross-sectional shape, volume, and taper on deposition profiles using a developmental AERx pulmonary delivery system. METHODS: Six mouthpieces were screened in the laboratory, and three were selected for clinical investigation: a cylindrical mouthpiece with constant-cross-sectional area, an elliptical mouthpiece with constant-cross-sectional area, and a tapered elliptical mouthpiece with an exit cross-sectional area equal to one half the entrance cross-sectional area. RESULTS: There was no significant difference in the lung dose or in the deposition pattern between males and females (p > 0.05, by ANOVA). The cross-sectional shape of the mouthpiece had no significant effect on the clinical lung dose or the deposition pattern (p > 0.05, by ANOVA), although in vitro testing showed lower emitted dose values with the tapered elliptical mouthpiece (by ANOVA and Duncan's multiple range test, alpha = 0.05). Using the tapered mouthpiece in the clinic resulted in significantly more deposition on the mouthpiece itself when compared to the nontapered mouthpieces. CONCLUSION: Inhalation of insulin using the AERx system was insensitive to differences in male and female respiratory tract geometry across all mouthpiece designs examined. PMID- 15553223 TI - Novel long-circulating lipid nanocapsules. AB - PURPOSE: To develop and evaluate novel long-circulating lipid nanocapsules (LN) designed for tumor delivery of lipophilic drugs. METHODS: Nanocapsules were produced by a solvent-free phase inversion process and were coated with polyethylene glycoldistearoylphosphatidylethanolamine conjugate (DSPE-PEG) during preparation or by a post-insertion step. In vivo studies were conducted in rats to assess LN pharmacokinetics and biodistribution. RESULTS: Post-insertion of DSPE-PEG appeared to be a convenient and effective method of obtaining LN of controlled sizes with high PEG density at their surface. After intravenous injection to rats, PEGylated lipid nanocapsules obtained by the post-insertion method exhibited long-circulating properties. Up to 50% of the injected dose was still present in the blood 8 h after administration for LN containing 6 mol% PEG 5000 or 10 mol% PEG 2000. This represented an area under the blood concentration time curve of almost 70% that of liposomes used in the Doxil formulation. CONCLUSION: With a simple solvent free-process, it was possible to produce long circulating LN of controlled sizes. Such LN could prove useful for the passive delivery of lipophilic anticancer drugs to solid tumors. PMID- 15553224 TI - Transforming growth factor (TGF)-beta1 releasing tricalcium phosphate/chitosan microgranules as bone substitutes. AB - PURPOSE: Tricalcium phosphate (TCP)/chitosan composite microgranules were developed as bone substitutes and tissue engineering scaffolds with the aim of obtaining a high bone forming efficacy. The microgranules have the ability to fill various types of defect sites with closer packing. In addition, the transforming growth factor-beta 1 (TGF-beta1) was added to the microgranules in order to improve bone-healing efficacy. METHODS: TCP/chitosan microgranules were fabricated by dropping a TCP suspended chitosan solution into a NaOH/ethanol solution. TGF-beta1 was incorporated into the TCP/chitosan microgranules by soaking the microgranules into the TGF-beta1 solution. Scanning electron microscopy (SEM) observations as well as experiments examining the release of TGF beta1 from chitosan and TCP/chitosan microgranules were performed. SEM was used to examine the cell morphologies on the microgranules, and the extent of cell proliferation was evaluated using a dimethyl-thiazol tetrazolium bromide (MTT) assay. The differentiated cell function was assessed by measuring the alkaline phosphatase activity as well as performing an osteocalcin assay. RESULTS: The size of the prepared microgranules was 350-500 microm and TCP powders were observed on the surface of the microgranules. TGF-beta1 was released from the TCP/chitosan microgranules at a therapeutic concentration for 4 weeks. The proliferation of osteoblasts on the TGF-beta1 loaded microgranules was the highest among the microgranules. SEM indicated that the seeded osteoblastic cells were firmly attached to the microgranules and proliferated in a multilayer fashion. The ALPase activity and osteocalcin content of all the samples increased during the culture period. CONCLUSIONS: These results suggest that the TCP/chitosan microgranules are potential bone substitutes with a drug releasing capacity and a osteoblastic cells culture scaffold. PMID- 15553225 TI - Retinal delivery of celecoxib is several-fold higher following subconjunctival administration compared to systemic administration. AB - PURPOSE: We have previously demonstrated that celecoxib, a selective COX-2 inhibitor, reaches the retina following repeated oral administrations and inhibits diabetes-induced vascular endothelial growth factor (VEGF) mRNA expression and vascular leakage in a rat model. The aim of this study was to quantify the relative retinal bioavailability of celecoxib from the subconjunctival route compared to a systemic route. METHODS: The plasma and ocular tissue distribution of celecoxib was determined in male Sprague-Dawley rats following subconjunctival and intraperitoneal administrations of drug suspension at a dose of 3 mg/rat. The animals were sacrificed at 0.5, 1, 2, 3, 4, 8, and 12 h post-dosing, the blood was collected, and the eyes were enucleated and frozen. The plasma, sclera, retina, vitreous, lens, and the cornea were isolated and celecoxib levels were determined using an HPLC method. The tissue exposure of the drug was measured as the area under the curve (AUC(0-infinity)) of the concentration vs. time profiles. The relative bioavailability was estimated as the AUC(0-infinity) ratio between subconjunctival and intraperitoneal groups. RESULTS: For the subconjunctivally dosed (ipsilateral) eye, the AUC(0-infinity) ratios between subconjunctival and intraperitoneal groups were 0.8 +/- 0.1, 53 +/- 4, 54 +/- 8, 145 +/- 21, 61 +/- 16, and 52 +/- 6 for plasma, sclera, retina, vitreous, lens, and cornea, respectively. For the contralateral ocular tissues, the AUC0-infinity ratios were 1.2 +/- 03, 11 +/- 0.3, 1.1 +/- 0.4, 1.0 +/- 0.3, and 1.2 +/- 0.3 in the sclera, retina, vitreous, lens, and the cornea, respectively, between the subconjunctival and the intraperitoneal groups. Assuming that the drug AUCs in contralateral eye were equal to the systemic pathway contribution to AUCs in the ipsilateral eye, the percent contribution of local pathways as opposed to systemic circulation for celecoxib delivery to the ipsilateral eye tissues was estimated to be 98% or greater. CONCLUSIONS: The retinal delivery of celecoxib was substantially higher following subconjunctival administration compared to the intraperitoneal route. The transscleral pathway almost completely accounts for the retinal celecoxib delivery following subconjunctival administration. PMID- 15553226 TI - Simultaneous extraction of urea and glucose by reverse iontophoresis in vivo. AB - PURPOSE: Reverse iontophoresis extracts glucose across the skin in the GlucoWatch Biographer, a device to monitor glycemia in diabetes. However, the device must first be calibrated with an invasive "fingerstick" and this has been perceived as a disadvantage. Here, urea, a neutral "internal standard" is extracted simultaneously in an attempt to render the technique completely non-invasive. METHODS: In a 5-h experiment in human volunteers, reverse iontophoretic fluxes of glucose and urea (J(glu) and J(urea), respectively) were measured periodically and correlated with the corresponding blood levels. In the case of glucose, a finger-tip blood sample was taken at the beginning of each collection interval; for urea, three blood samples were assayed: one before, one during, and one at the end of iontophoresis. RESULTS: The ratio J(glu)/J(urea) divided by the ratio of the systemic concentrations (C(glu)/C(urea)) yielded an extraction coefficient (K) that could be compared between subjects. Though J(glucose) tracked C(glu) faithfully when the volunteers were challenged with an oral glucose load, J(urea) remained quite stable reflecting the fact that C(urea) did not change appreciably during the experiment. However, whereas the variability (expressed as the coefficient of variation) in the normalized extraction flux of urea (J(urea)/C(urea)) was on the order of 25%, that for glucose was greater (>45%), with the result that the values of K (0.45 +/- 0.25) were less constant than anticipated. CONCLUSIONS: Although urea performed quite reasonably as an internal standard, in that its extraction flux and systemic concentration both remained quite constant, the normalized, transdermal, iontophoretic flux of glucose showed interindividual variability due to mechanisms that were not entirely governed by electrotransport. That is, despite good qualitative tracking to blood levels, there appear to be other (biochemical, metabolic, contamination?) factors that impact upon the quantitative results obtained. PMID- 15553227 TI - Influence of spinline stress on release properties of a coaxial controlled release device based on EVA polymers. AB - PURPOSE: To investigate the influence of the extrusion parameters on the polymeric structure and release properties of polyethylene vinyl acetate (EVA) coaxial fibers, used for controlled release of steroids. METHODS: Coaxial fibers were prepared under various extrusion conditions. Both spinline stress and release properties were determined. The polymeric structure of the membrane was investigated with wide angle X-ray scattering (WAXS). RESULTS: Upon leaving the spinneret, the polymeric fiber exhibits a large die swell. As a consequence, it is necessary to apply a force to draw the fiber to its desired diameter. A larger drawing force is needed at lower extrusion temperature, at a smaller air gap, or at a higher spinning velocity. It was found that the release rate of a steroid from the coaxial fiber increases, when the fibers are prepared at a higher spinline stress. X-ray measurements reveal that at higher spinline stress, the crystalline volume fraction of the membrane decreases. As a result of a decreasing crystallinity, the permeability of the polymer increases. CONCLUSIONS: It is demonstrated that the extrusion parameters and spinline stress have a significant influence on the polymeric structure of the membrane and hence the release properties. Higher spinline stress results in a higher release rate. PMID- 15553228 TI - Enhancement of topical delivery from biodegradable nanoparticles. AB - PURPOSE: To determine whether and how encapsulation of lipophilic compounds in polymeric nanoparticles is able to improve topical delivery to the skin. METHODS: The penetration of octyl methoxycinnamate (OMC; Parsol MCX), a highly lipophilic sunscreen, into and across porcine ear skin in vitro was investigated, subsequent to encapsulation in poly(epsilon-caprolactone) nanoparticles, using tape stripping. Confocal laser scanning microscopy (CLSM) was used to visualize the distribution of nanoparticles, charged with Nile red (NR), a lipophilic and fluorescent dye. RESULTS: Quantification of OMC in the skin using tape-stripping demonstrated that nanoparticulate encapsulation produced a 3.4-fold increase in the level of OMC within the stratum corneum (SC), although the use of nanoparticles did not appear to increase skin permeation (it was not possible to detect OMC in the receiver compartment after 6 h). The confocal images showed that the fluorescence profile observed in the skin after application of NR containing nanoparticles was clearly different from that seen following application of NR dissolved in propylene glycol. Two hours postapplication of NR containing nanoparticles, fluorescence was perceptible at greater depths (up to 60 microm) within the skin. CONCLUSIONS: i) Nanoparticulate encapsulation of OMC increased its "availability" with the SC. ii) The altered distribution of NR when delivered via nanoparticles was due, at least in part, to its altered thermodynamic activity (relative to that in propylene glycol) and, as a result, an increase in its partition coefficient into the SC. PMID- 15553229 TI - The cohesive-adhesive balances in dry powder inhaler formulations II: influence on fine particle delivery characteristics. AB - PURPOSE: To investigate the influence of the cohesive-adhesive balances on dry powder formulation aerosolization and delivery characteristics. METHODS: De agglomeration properties of pharmaceutical powders were investigated using an Aerosizer at various shear forces. Aerosol drug deposition properties of drug only formulations and carrier-based formulations were investigated using a low resistance device (Rotahaler) and a high-resistance device (Turbuhaler) via a twin-stage impinger. RESULTS: A paradoxical relationship between particle cohesive strength and de-agglomeration efficiencies of drug-only formulations was observed, where an increase in cohesive strength led to a higher fine particle fraction. A possible explanation for the variation in the fluidization and aerosolization properties between low and high cohesive particles was modeled on the relationship between cohesion, metastable agglomerate size, and the resulting aerodynamic drag force acting on the fluidized agglomerates. The addition of a fine particle lactose carrier influenced the drug deposition patterns in different ways depending on the relative cohesive and adhesive force balances within the formulation. CONCLUSIONS: The use of the colloid Atomic Force Microscrope (AFM) technique in combination with the cohesive-adhesive balance (CAB) system provides a novel preformulation tool for investigating the likely behavior of a dry powder formulation and a possible means of interpreting the possible de-aggregation and dispersion mechanisms of carrier-based formulations. PMID- 15553230 TI - Supercooled smectic nanoparticles: a potential novel carrier system for poorly water soluble drugs. AB - PURPOSE: The possibility of preparing nanoparticles in the supercooled thermotropic liquid crystalline state from cholesterol esters with saturated acyl chains as well as the incorporation of model drugs into the dispersions was investigated using cholesteryl myristate (CM) as a model cholesterol ester. METHODS: Nanoparticles were prepared by high-pressure melt homogenization or solvent evaporation using phospholipids, phospholipid/ bile salt, or polyvinyl alcohol as emulsifiers. The physicochemical state and phase behavior of the particles was characterized by particle size measurements (photon correlation spectroscopy, laser diffraction with polarization intensity differential scattering), differential scanning calorimetry, X-ray diffraction, and electron and polarizing light microscopy. The viscosity of the isotropic and liquid crystalline phases of CM in the bulk was investigated in dependence on temperature and shear rate by rotational viscometry. RESULTS: CM nanoparticies can be obtained in the smectic phase and retained in this state for at least 12 months when stored at 230C in optimized systems. The recrystallization tendency of CM in the dispersions strongly depends on the stabilizer system and the particle size. Stable drug-loaded smectic nanoparticles were obtained after incorporation of 10% (related to CM) ibuprofen, miconazole, etomidate, and 1% progesterone. CONCLUSIONS: Due to their liquid crystalline state, colloidal smectic nanoparticles offer interesting possibilities as carrier system for lipophilic drugs. CM nanoparticles are suitable model systems for studying the crystallization behavior and investigating the influence of various parameters for the development of smectic nanoparticles resistant against recrystallization upon storage. PMID- 15553231 TI - The potential of Raman spectroscopy as a process analytical technique during formulations of topical gels and emulsions. AB - PURPOSE: The primary objective of this study is to investigate the possibility of using Raman spectroscopy as a process analytical technique (PAT) for quality control during manufacturing of topical dosage forms. METHODS: A fiber-optic Raman probe was used to monitor the formulation of pharmaceutical gel and emulsion in laboratory scale. Raman shifts of typical commercial raw materials used in topical dosage forms were measured to ascertain the potential of this technique for monitoring and analyzing topical products. Spectra of some well characterized topical gels manufactured in our laboratory were also measured. RESULTS: Commercial raw materials were found to be Raman sensitive. Due to the difference in chemical composition, raw materials exhibit characteristic peaks that can be exploited to monitor formulation processes. Spectra taken during formulation of an emulsion using Carbopol Ultrez as thickener and Tefose as emulsifying agent show changes in Raman shifts immediately after major formulation steps. CONCLUSIONS: The findings from this work suggest that Raman spectroscopy can be a valuable process analytical technique for quality control of topical gel and cream formulations. PMID- 15553232 TI - Amphiphilogels for drug delivery: formulation and characterization. AB - PURPOSE: This study examines the microstructure, gelation temperatures, and flow properties of novel amphiphilogels consisting solely of non-ionic surfactants. METHODS: Gels were prepared by mixing the solid gelator (sorbitan monostearate or sorbitan monopalmitate) and the liquid phase (liquid sorbitan esters or polysorbates) and heating them at 60 degrees C to form a clear isotropic sol phase, and cooling the sol phase to form an opaque semisolid at room temperature. Gel microstructure was examined by light and electron microscopy and by small angle neutron scattering (SANS); gelation temperatures were measured by hotstage microscopy, a melting point apparatus, and high sensitivity differential scanning calorimetry (HSDSC). Flow rheograms were performed to establish the zero-rate viscosity of the gels and their performance under shear. RESULTS: Gel microstructures consisted mainly of clusters of tubules of gelator molecules that had aggregated upon cooling of the sol phase, forming a 3D network throughout the continuous phase. The gels demonstrated thermoreversibility. Gelation temperature and viscosity increased with increasing gelator concentration, indicating a more robust gel network. At temperatures near the skin surface temperature, the gels softened considerably; this would allow topical application. CONCLUSIONS: This study has demonstrated the formation/preparation of stable, thermoreversible, thixtropic surfactant gels (amphiphilogels) with suitable physical properties for topical use. PMID- 15553233 TI - Comparative effects on intestinal absorption in situ by P-glycoprotein-modifying HIV protease inhibitors. AB - PURPOSE: P-glycoprotein (P-gp) is made responsible for the limited oral bioavailability of P-gp substrates like peptidic HIV protease inhibitors (PIs). With respect to combined application of two PIs in antiretroviral regimes, we first investigated the influences on intestinal saquinavir uptake using different PIs in in situ perfusion studies. METHODS: Perfusion experiments were carried out in three intestinal segments with P-gp substrates talinolol and saquinavir using fixed concentrations of PIs and with each varying concentrations in the jejunum and ileum. Furthermore, cellular uptake of fluorescent P-gp substrate rhodamine 123 and MRP-substrate carboxyfluorescein has each been quantified in P-gp and MRP expressing cells by flow cytometry under co-administration of PIs. RESULTS: Increase of calculated permeabilities of P-gp-specific substrate talinolol was found under co-administration of both PIs, ritonavir and H17, with highest absorption rates in the ileal and colon segment. H17 proved to be a better P-gp inhibitor than ritonavir by resulting IC50 values and also in the cellular uptake of rhodamine. Similar increases of permeabilities in ileum and colon have also been found for saquinavir as P-gp as well as MRP-substrate with differences in the jejunal uptake, which was found higher for H17. Additional MRP-inhibitory activities of H17 were proved by increasing cellular uptake rates of carboxyfluorescein in MRP-expressing cells. CONCLUSIONS: The investigated PIs were characterized as effective P-gp inhibitors in the intestinal absorption of P gp substrates. H17 showed MRP-inhibitory effects that also favor intestinal drug absorption of corresponding substrates. With respect to combined therapeutic application of PIs, compounds like H17 raise hopes for improved bioavailability of poorly absorbed compounds. PMID- 15553234 TI - Use of loperamide as a phenotypic probe of mdr1a status in CF-1 mice. AB - PURPOSE: A subpopulation of the CF-1 mouse strain (approximately 25%) lacks P-gp expression, and consequently, increased brain penetration of many substrates is observed in these animals. Mice lacking the mdr1a gene represent an important research tool to study the potential effects of P-gp on CNS substrate disposition. METHODS: Adult CF-1 mice were used in all experiments. Loperamide induced antinociception was determined by the hotplate latency test at 0.25, 2, and 4 h post-dose. At the conclusion of the pharmacodynamic experiment(s), trunk blood and brain tissue were collected and analyzed by high-performance liquid chromatography-mass spectrometry (LC-MS/MS). Mice were also genotyped for their mdrla status via RT-PCR. RESULTS: All mice with three consecutive effects of maximum hotplate latency (60 s) showed considerable opioid-like behavior in addition to antinociception. Mice without three consecutive effects of maximum hotplate latency (< or = 30 s) showed no opioid-like behavior. The loperamide brain-to-serum ratio in mice identified as P-gp-deficient was 65-fold higher compared to the P-gp-competent animals (10.1 +/- 1.0 vs. 0.155 +/- 0.018). All animals identified as phenotypically P-gp-competent based on the hotplate assay evidenced the mdrla(+/+) genotype. CONCLUSION: This assay appears to offer a rapid and unambiguous measure via a relatively non-invasive, simple technique to identify P-gp status in the CF-1 subpopulation of mice. PMID- 15553235 TI - Phototoxicity of 5-aminolevulinic acid in the HeLa cell line as an indicative measure of photodynamic effect after topical administration to gynecological lesions of intraepithelial form. AB - PURPOSE: The depth-resolved pattern of aminolevulinic acid (ALA) concentration in excised vaginal tissue was determined after in vitro application of an ALA-loaded bioadhesive patch. From this data, the tissue concentration of ALA achievable at a specified depth from the surface could be related to the concentration needed to elicit a photodynamic effect in a model gynecological tumor cell line (HeLa). METHODS: Excised vaginal tissue was mounted in a modified Franz diffusion cell and exposed to a water-soluble, ALA-loaded, bioadhesive patch. After a period of time, the tissue was cryostatically sectioned and the stratal concentration of radiolabeled ALA determined using scintillation spectroscopy. HeLa cells were cultured in media containing specific concentrations of ALA and exposed to standard photodynamic protocols of light exposure. RESULTS: An ALA concentration of 65.6 mM was achievable at 2.375 mm from the tissue surface after application of ALA-loaded patch. The photodynamic effectiveness of this concentration was demonstrated in HeLa with exposure to concentrations exceeding 1.0 mM ALA bringing about reductions in viable cell numbers by 90%. An enhancement of PpIX production using adjunctive EDTA over the clinically relevant 4 h application time interval was shown to be minimal in HeLa. Instead, PpIX production was more closely correlated with ALA concentration, with 100 mM ALA producing approximately 3100 ng PpIX mg(-1) protein in the same time period. CONCLUSIONS: Given that vaginal intraepithelial neoplasias can extend to 2.0 mm from the lesion surface, the ALA permeability derived from a bioadhesive patch is sufficient to induce photosensitization suitable for light induced destruction at deep sites of this type of lesion. PMID- 15553236 TI - Enhancement of the efficacy of an antagonist of an extracellular receptor by attachment to the surface of a biocompatible carrier. AB - PURPOSE: In order to improve the in vitro and in vivo efficacy of an integrin antagonist (IA) of the extracellular domain of the alphavbeta3 integrin, a receptor upregulated on tumor neovasculature, the IA was attached to the surface of a dextran-coated liposome (DCL). IA-DCLs were characterized in vitro, and the pharmacokinetic and antitumor properties were assessed in vivo. METHODS: The in vitro binding properties were measured with purified integrin, endothelial cells, and melanoma cells. The pharmacokinetic parameters were measured in healthy mice with 14C-labeled IA-DCLs and anti-tumor efficacy was assessed with the M21 human melanoma xenograft mouse model. RESULTS: In vitro, IC50 values for IA-DCLs and IA are similar, and IA-DCLs inhibit cell proliferation relative to controls. IA-DCLs are stable in serum, and the pharmacokinetic half-life in mice is 23 h. In the M21/mouse model, statistically significant inhibition of tumor growth was observed for mice treated with IA-DCLs, whereas controls including saline, DCLs lacking IA, and cyclo(RGDfV) were ineffective. Increased apoptosis and a reduction in vessel counts relative to controls were present in tumors from animals treated with IA-DCLs. CONCLUSIONS: These results demonstrate that IA-DCLs are potent anti-angiogenic therapeutic agents with superior in vivo activity and pharmacology compared to unmodified IA. PMID- 15553237 TI - Screening of the interaction between xenobiotic transporters and PDZ proteins. AB - PURPOSE: Xenobiotic transporters have been proposed to be involved in membrane penetration of various therapeutic agents. As little information is available on molecular mechanism of their functional regulation, we have attempted to clarify the protein-protein interactions of such transporters as a first step to identify their regulators. METHODS: Yeast two-hybrid screening was performed to examine the interaction between carboxylic terminus of various xenobiotic transporters and PDZ (PSD95, D1g and ZO1) domain-containing proteins. The interaction and functional regulation were also evaluated in pull-down, immunoprecipitation and transport studies. RESULTS: Specific interaction with PDZ proteins was identified for several xenobiotic transporters including PEPT1, PEPT2, OCT3, OCTN1, OCTN2, OAT4, OATP-A, OATP-D, and OATP-F. The potent interaction was observed between PEPT2 and PDZK1, and deletion of the last four amino acids of the PEPT2 C terminus almost completely abrogated such interaction. Recombinant PEPT2 C terminus fusion protein can bind to purified His6-tagged PDZK1, confirming the involvement of two of four PDZ domains within PDZK1 in the interaction. Alanine scanning mutation in PEPT2 revealed the presence of a consensus sequence (-T-X-L) that is responsible for the PDZK1 interaction. Transfection of PDZK1 increased the uptake of glycylsarcosine by PEPT2, whereas such stimulation was not observed for PEPT2 with the last four amino acids deleted. CONCLUSIONS: These results first identified the interaction between PDZ proteins and the cytosolic tail of various xenobiotic transporters. PDZK1 directly interacts with PEPT2, exerting functional regulation of its transporting activity. The current findings imply the localization of PEPT2 within a protein network constructed from PDZK1 and other transporter proteins. PMID- 15553238 TI - Functional analysis of SNPs variants of BCRP/ABCG2. AB - PURPOSE: The aim of the current study was to identify the effect of single nucleotide polymorphisms (SNPs) in breast cancer resistance protein (BCRP/ABCG2) on its localization, expression level, and transport activity. METHODS: The cellular localization was identified using the wild type and seven different SNP variants of BCRP (V12M, Q141K, A149P, R163K, Q166E, P269S, and S441N BCRP) after transfection of their cDNAs in plasmid vector to LLC-PK1 cells. Their expression levels and transport activities were determined using the membrane vesicles from HEK293 cells infected with the recombinant adenoviruses containing these kinds of BCRP cDNAs. RESULTS: Wild type and six different SNP variants of BCRP other than S441N BCRP were expressed on the apical membrane, whereas S441N BCRP showed intracellular localization. The expression levels of Q141K and S441N BCRP proteins were significantly lower compared with the wild type and the other five variants. Furthermore, the transport activity of E1S, DHEAS, MTX, and PAH normalized by the expression level of BCRP protein was almost the same for the wild type, V12M, Q141K, A149P, R163K, Q166E, and P269S BCRP. CONCLUSIONS: These results suggest that Q141K SNPs may associate with a lower expression level, and S441N SNPs may affect both the expression level and cellular localization. It is possible that subjects with these polymorphisms may have lower expression level of BCRP protein and, consequently, a reduced ability to export these substrates. PMID- 15553239 TI - Effects of extracts of commonly consumed food supplements and food fractions on the permeability of drugs across Caco-2 cell monolayers. AB - PURPOSE: Extracts made from berries, herbs, and various plant materials, which might possess a range of activities, are used as health promoting products. Because little is known about their effects on the absorption of co-administered drugs, the effects of some food supplements, Finnish berries, and herbs were studied on the permeability of some commonly used drugs. METHODS: The permeabilities of verapamil, metoprolol, ketoprofen, paracetamol, and furosemide were studied across Caco-2 cell monolayers with contemporaneously administered extracts from flax seed, purple loosestrife, and Scots pine bark; bilberries, cowberries, and raspberries; oregano, rosemary, and sage. Toxicological tests were conducted to determine cellular damage. RESULTS: The effects of extracts on drug permeabilities were generally minor. Flax seed decreased the permeability of all drugs except verapamil. Purple loosestrife and pine decreased verapamil and metoprolol permeability. Changes caused by berries were mainly pH-related. Rosemary and oregano enhanced furosemide permeability. CONCLUSIONS: Ingestion of extracts of herbs and berries studied are not expected to markedly change the permeabilities of highly permeable drugs. Harmful effects at sites of or during absorption are unlikely. However, if high doses of extracts are administered with low permeable drugs in vitro, effects on drug permeabilities could not be excluded. Use of such extracts should therefore be evaluated during continuous medication. PMID- 15553240 TI - Binding and uptake of wheat germ agglutinin-grafted PLGA-nanospheres by caco-2 monolayers. AB - PURPOSE: The Caco-2 association of lectin-grafted PLGA-nanospheres was investigated compared to plain and BSA-coated spheres. METHODS: Nanospheres made from fluorescent-labeled PLGA were coated with wheat germ agglutinin (WGA) or BSA and incubated with Caco-2 monolayers varying the concentration of nanospheres, the time, and the temperature. The tests were performed in a static horizontal as well as an aerated vertical setup to find out the system most appropriate for estimation of bioadhesion. RESULTS: Due to bioadhesive effects, WGA-modified particles exhibited highest association to the cells as compared to plain and BSA coated ones. The amount of associated spheres increased with time and concentration of the nanosphere suspension. Whereas the binding of lectin-coated spheres was independent from energy, their uptake was energy consuming as opposed to BSA and plain nanospheres, which exhibited nonspecific, energy independent binding and uptake. Although more particles were associated with the monolayer in the horizontal setup than in the vertical system, the vertical system reflects true bioadhesion due to circulation of the spheres which inhibits the influence of sedimentation. CONCLUSIONS: Immobilization of WGA considerably enhances the binding as well as the uptake of PLGA-nanospheres by Caco-2 monolayers. For bioadhesion studies, the vertical setup is recommended instead of the horizontal setup. PMID- 15553241 TI - Functional analysis of recombinant human serum albumin domains for pharmaceutical applications. AB - PURPOSE: Functional analysis of the three recombinant human serum albumin (rHSA) domains and their potential as stand-alone proteins for use as drug delivery protein carriers. METHODS: Protein structure was examined by fluorescence and CD spectroscopy. Ligand binding was estimated by ultrafiltration. Antioxidant activity was estimated by measuring the quenching of dihydrorhodamine 123. Esterase-like activity and enolase-like activity were estimated from the rate of hydrolysis of p-nitrophenyl acetate and conversion of dihydrotestosterone from the 3-keto to 3-enol form, respectively. The domains of human serum albumin (HSA) were radiolabeled with 111In to evaluate their pharmacokinetics. RESULTS: The ligand binding ability of subsites Ia and Ib could not be detected in domain II. However, the binding of ligands to subsite Ic and site II were preserved in domain II and domain III, respectively. Domain III retained about 45% of its esterase-like activity, and weaker esterase-like activity was also observed in domain I. All domains showed low enolase-like activity in a pH 7.4 phosphate buffer, but domain II had higher activity in a pH 9.2 carbonate buffer. Domain I exhibited antioxidant activity comparable to that of rHSA. All three of the 111In radiolabeled domains were rapidly eliminated from HSA, with high accumulation in the kidneys. CONCLUSION: Domain I of HSA has great potential for further development as a drug delivery protein carrier, due to its favorable properties and the presence of a free cysteine residue. PMID- 15553242 TI - Novel scaled average bioequivalence limits based on GMR and variability considerations. AB - PURPOSE: i) To develop novel approaches for the construction of bioequivalence (BE) limits incorporating both the intrasubject variability and the geometric mean ratio (GMR), and ii) to assess the performance of the novel approaches in comparison to several scaled BE procedures and the classic unscaled average BE. METHODS: Plots of the BE limits or the extreme GMR values accepted as a function of the coefficient of variation (CV) were constructed for published and the developed scaled procedures. Two-period crossover BE investigations with 12, 24, or 36 subjects were simulated with assumptions of a CV 10%, 20%, 30%, or 40%. The decline in the percentage of accepted studies was recorded as the true GMR for the two formulations was raised from 1.00 to 1.50. Acceptance of BE was evaluated by published and the developed scaled procedures, and, for comparison, by the unscaled average BE. RESULTS: Two GMR-dependent BE limits are proposed for the evaluation of average BE: i) BELscG1 with Ln(Upper, Lower BE limit) = +/-[(5 - 4GMR)0.496s + Ln(1.25)], and ii) BELscG2 with Ln(Upper, Lower BE limit) = +/-[(3 2GMR)(0.496s + Ln(1.25))], where s is the square root of the intrasubject variance. The range of BE limits becomes narrower as GMR values deviate from unity, and increases with variability. The two new approaches exhibit the highest statistical power at low CV values. At high levels of variability, BELscG1 and BELscG2 show high statistical power, as well as the lowest percentages of acceptance among the scaled methods when GMR = 1.25. The latter becomes more obvious when a large number of subjects is incorporated in the studies. CONCLUSIONS: The GMR and CV estimates of the BE study can be used in conjunction with the GMR vs. CV plot for the assessment of average BE. The new approaches, BELscG1 and BELscG2, appear to be highly effective at all levels of variation investigated. PMID- 15553243 TI - Distribution of phytoplasmas in infected plants as revealed by real-time PCR and bioimaging. AB - Phytoplasmas are cell wall-less bacteria inhabiting the phloem and utilizing it for their spread. Infected plants often show changes in growth pattern and a reduced crop yield. A quantitative real-time polymerase chain reaction (Q-PCR) assay and a bioimaging method were developed to quantify and localize phytoplasmas in situ. According to the Q-PCR assay, phytoplasmas accumulated disproportionately in source leaves of Euphorbia pulcherrima and, to a lesser extent, in petioles of source leaves and in stems. However, phytoplasma accumulation was small or nondetectable in sink organs (roots and sink leaves). For bioimaging, infected plant tissue was stained with vital fluorescence dyes and examined using confocal laser scanning microscopy. With a DNA-sensitive dye, the pathogens were detected exclusively in the phloem, where they formed dense masses in sieve tubes of Catharanthus roseus. Sieve tubes were identified by counterstaining with aniline blue for callose and multiphoton excitation. With a potentiometric dye, not all DNA-positive material was stained, suggesting that the dye stained metabolically active phytoplasmas only. Some highly infected sieve tubes contained phytoplasmas that were either inactive or dead upon staining. PMID- 15553244 TI - Role of chemotaxis toward fusaric acid in colonization of hyphae of Fusarium oxysporum f. sp. radicis-lycopersici by Pseudomonas fluorescens WCS365. AB - Pseudomonas fluorescens WCS365 is an excellent competitive colonizer of tomato root tips after bacterization of seed or seedlings. The strain controls tomato foot and root rot (TFRR) caused by the phytopathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici. Under biocontrol conditions, fungal hyphae were shown to be colonized by WCS365 bacteria. Because chemotaxis is required for root colonization by WCS365 cells, we studied whether chemotaxis also is required for hyphae colonization. To that end, an in vitro assay was developed to study hyphae colonization by bacteria. The results indicated that cells of the cheA mutant FAJ2060 colonize hyphae less efficiently than cells of wild-type strain WCS365, when single strains were analyzed as well as when both strains were applied together. Cells of WCS365 show a chemotactic response toward the spent growth medium of F. oxysporum f. sp. radicis-lycopersici, but those of its cheA mutant, FAJ2060, did not. Fusaric acid, a secondary metabolite secreted by Fusarium strains, appeared to be an excellent chemo-attractant. Supernatant fluids of a number of Fusarium strains secreting different levels of fusaric acid were tested as chemo-attractants. A positive correlation was found between chemo-attractant activity and fusaric acid level. No chemotactic response was observed toward the low fusaric acid-producer FO242. Nevertheless, the hyphae of FO242 still were colonized by WCS365, suggesting that other metabolites also play a role in this process. The possible function of hyphae colonization for the bacterium is discussed. PMID- 15553245 TI - Diverse members of the AvrBs3/PthA family of type III effectors are major virulence determinants in bacterial blight disease of rice. AB - AvrXa7 is a member of the avBs3/pthA gene family and the only known type III secretion system effector gene from Xanthomonas oryzae pv. oryzae with a major contribution to bacterial growth and lesion formation in bacterial blight disease of rice. We examined the general requirement for effectors of the AvrBs3/PthA family in bacterial blight of rice by identifying effectors from diverse strains of the pathogen. Inactivation of single effector genes in representative strains from Japan, Korea, and the Philippines resulted in severely limited growth in plants. Five strains harbored one gene of the avrBs3/pthA family, while one strain had two genes with the equivalent virulence activity of avrXa7. Sequence analysis revealed three genes with unique repeat arrangements in comparison to avrXa7. Comparison of the repetitive regions revealed a potential motif for the group that was also present in the repetitive region of avrBs3. However, the repetitive region of avrBs3 could not support virulence activity but, in combination with the C-terminal coding region of avrXa7, triggered a Xa7 dependent avirulence reaction. The results revealed diverse members of the avrBs3/pthA gene family with virulence activity in X. oryzae pv. oryzae and supported the hypothesis that bacterial blight disease of rice is highly dependent on a single class of type III effectors. The results also indicated that avrXa7 avirulence specificity is separable from virulence activity. PMID- 15553247 TI - cDNA macroarray analysis of gene expression in ineffective nodules induced on the Lotus japonicus sen1 mutant. AB - The Lotus japonicus sen1 mutant forms ineffective nodules in which development is arrested at the stage of bacterial differentiation into nitrogen-fixing bacteroids. Here, we used cDNA macroarray systems to compare gene expression in ineffective nodules induced on the sen1 mutant with gene expression in wild-type nodules, in order to identify the host plant genes that are involved in nitrogen fixation. Macroarray analysis coupled with Northern blot analysis revealed that the expression of 18 genes was significantly enhanced in ineffective sen1 nodules, whereas the expression of 30 genes was repressed. Many of the enhanced genes encoded hydrolase enzymes, such as cysteine proteinase and asparaginase, that might function in the early senescence of sen1 nodules. By contrast, the repressed genes encoded nodulins, enzymes that are involved in carbon and nitrogen metabolism, membrane transporters, enzymes involved in phytohormone metabolism and secondary metabolism, and regulatory proteins. These proteins might have a role in the establishment of nitrogen fixation. In addition, we discovered two novel genes that encoded glutamate-rich proteins and were localized in the vascular bundles of the nodules. The expression of these genes was repressed in the ineffective nodules, which had lower levels of nitrogenase activity. PMID- 15553246 TI - Identification and expression profiling of tomato genes differentially regulated during a resistance response to Xanthomonas campestris pv. vesicatoria. AB - The gram-negative bacterium Xanthomonas campestris pv. vesicatoria is the causal agent of spot disease in tomato and pepper. Plants of the tomato line Hawaii 7981 are resistant to race T3 of X. campestris pv. vesicatoria expressing the type III effector protein AvrXv3 and develop a typical hypersensitive response upon bacterial challenge. A combination of suppression subtractive hybridization and microarray analysis identified a large set of cDNAs that are induced or repressed during the resistance response of Hawaii 7981 plants to X. campestris pv. vesicatoria T3 bacteria. Sequence analysis of the isolated cDNAs revealed that they correspond to 426 nonredundant genes, which were designated as XRE (Xanthomonas-regulated) genes and were classified into more than 20 functional classes. The largest functional groups contain genes involved in defense, stress responses, protein synthesis, signaling, and photosynthesis. Analysis of XRE expression kinetics during the tomato resistance response to X. campestris pv. vesicatoria T3 revealed six clusters of genes with coordinate expression. In addition, by using isogenic X. campestris pv. vesicatoria T2 strains differing only by the avrXv3 avirulence gene, we found that 77% of the identified XRE genes were directly modulated by expression of the AvrXv3 effector protein. Interestingly, 64% of the XRE genes were also induced in tomato during an incompatible interaction with an avirulent strain of Pseudomonas syringae pv. tomato. The identification and expression analysis of X. campestris pv. vesicatoria T3-modulated genes, which may be involved in the control or in the execution of plant defense responses, set the stage for the dissection of signaling and cellular responses activated in tomato plants during the onset of spot disease resistance. PMID- 15553248 TI - Cloning, characterization, and evolution of the NBS-LRR-encoding resistance gene analogue family in polyploid cotton (Gossypium hirsutum L.). AB - The nucleotide-binding site-leucine-rich repeat (NBS-LRR)-encoding gene family has attracted much research interest because approximately 75% of the plant disease resistance genes that have been cloned to date are from this gene family. We cloned the NBS-LRR-encoding genes from polyploid cotton by a polymerase chain reaction-based approach. A sample of 150 clones was selected from the NBS-LRR gene sequence library and was sequenced, and 61 resistance gene analogs (RGA) were identified. Sequence analysis revealed that RGA are abundant and highly diverged in the cotton genome and could be categorized into 10 distinct subfamilies based on the similarities of their nucleotide sequences. The numbers of members vary many fold among different subfamilies, and gene index analysis showed that each of the subfamilies is at a different stage of RGA family evolution. Genetic mapping of a selection of RGA indicates that the RGA reside on a limited number of the cotton chromosomes, with those from a single subfamily tending to cluster and two of the RGA loci being colocalized with the cotton bacterial blight resistance genes. The distribution of RGA between the two subgenomes A and D of cotton is uneven, with RGA being more abundant in the A subgenome than in the D subgenome. The data provide new insights into the organization and evolution of the NBS-LRR-encoding RGA family in polyploid plants. PMID- 15553249 TI - Mutations in wheat exhibiting growth-stage-specific resistance to biotrophic fungal pathogens. AB - Two mutants were isolated in wheat that showed enhanced resistance towards Puccinia striiformis f. sp. tritici, the fungal causal agent of yellow rust. The altered phenotype of I3-48 is due to a minimum of two mutation events, each showing a partial, additive effect, with one mutation segregating with a deletion on the long arm of chromosome 4D. In the case of I3-54, the enhanced resistance is due to a single, dominant mutation. In both mutants, the expression of the enhanced resistance is growth-stage specific. With I3-54, the full resistance phenotype is apparent from the third seedling leaf onwards, while with I3-48, a full resistance phenotype is only seen on the tenth and subsequent leaves. In addition to the enhanced resistance towards yellow rust, I3-48 also shows enhanced resistance towards brown rust, and I3-54 shows enhanced resistance to powdery mildew. PMID- 15553250 TI - Transcriptional regulation of components of the type III secretion system and effectors in Pseudomonas syringae pv. phaseolicola. AB - Quantitative real-time polymerase chain reaction was used with specific TaqMan probes to examine transcription of selected hrp and effector genes in Pseudomonas syringae pv. phaseolicola strains 1448A (race 6) and 1449B (race 7). Transcripts examined were from genes encoding the regulators hrpR and hrpL, core structural components of the type III secretion system (TTSS) hrcC, hrcJ, hrcN, hrcU, and hrpA; the first open-reading frame of each hrp operon, including hrpF, hrpJ, hrpP, and hrpY, and also secreted effectors hrpZ, avrPphE, avrPphF, and virPphA. All genes were induced by incubation in a minimal medium and showed patterns of expression indicating regulation by HrpRS and HrpL. Basal mRNA levels and the timing of accumulation of transcripts after induction differed significantly, suggesting the operation of additional regulatory elements. However, no clear transcriptional hierarchy emerged to explain the ordered construction of the TTSS. Quantitative analysis confirmed that the rates and levels of transcript accumulation within the first 2 h after inoculation were considerably higher in planta than in vitro, and indicated that plant cell wall contact may enhance transcription of TTSS and effector genes in P. syringae pv. phaseolicola. The low abundance hrcU mRNA had a half-life of 16.5 min, whereas other transcripts had half-lives between 3 and 8 min. PMID- 15553251 TI - NopB, a soybean cultivar-specificity protein from Sinorhizobium fredii USDA257, is a type III secreted protein. AB - The type III secretion system (TTSS) of plant- and animal-pathogenic bacteria is involved in translocation of virulence factors into the host cell cytosol where they modulate cellular processes. Sinorhizobium fredii USDA257 is a gram-negative soil bacterium that forms nitrogen-fixing nodules on specific soybean cultivars (Glycine max (L.) Merr.). This microsymbiont is known to secrete at least five nodulation outer proteins (Nops) in response to flavonoid induction. Some of these Nops have been shown to be secreted by TTSS in this symbiotic bacterium. We have isolated and purified an 18-kDa extracellular protein from flavonoid-induced cultures of USDA257. The N-terminal amino acid sequence of this purified protein was identical to the published sequence of the soybean cultivar-specificity protein, NopB (formerly NoIB). Inactivation of rhcN, which encodes an ATPase, abolished secretion of NopB. Similarly, a nonpolar nopB deletion mutant was compromised in its ability to secrete several Nops. A construct containing the coding region of nopB under control of a T7 promoter was expressed successfully in Escherichia coli and, subsequently, the recombinant NopB was purified by nickel-affinity column chromatography. Polyclonal antibodies raised against purified recombinant NopB were used in Western blot analysis to demonstrate the association of NopB with pilus-like surface appendages. Deletion analysis indicated that the first 33 N-terminal residues of NopB were necessary and sufficient to mediate the secretion of a green fluorescent protein reporter. Introduction of plasmid-borne extra copies of nopB into USDA257 resulted in lower accumulation of native NopB. We also show that USDA257 and its nonpolar nopB deletion mutant exhibited discernible differences in their ability to nodulate legume hosts. PMID- 15553252 TI - Involvement of N-acylhomoserine lactones throughout plant infection by Erwinia carotovora subsp. atroseptica (Pectobacterium atrosepticum). AB - Erwinia carotovora subsp. atroseptica is responsible for potato blackleg disease in the field and tuber soft rot during crop storage. The process leading to the disease occurs in two phases: a primary invasion step followed by a maceration step. Bacteria-to-bacteria communication is associated with a quorum-sensing (QS) process based on the production of N-acylhomoserine lactones (HSL). The role of HSL throughout plant infection was analyzed. To this purpose, HSL produced by a specific E. carotovora subsp. atroseptica wild-type strain, which was particularly virulent on potato, were identified. A derivative of this strain that expressed an HSL lactonase gene and produced low amounts of HSL was generated. The comparison of these strains allowed the evaluation of the role of HSL and QS in disease establishment and development. Bacterial growth and motility; activity of proteins secreted by type I, II, and III systems; and hypersensitive and maceration reactions were evaluated. Results indicated that HSL production and QS regulate only those traits involved in the second stage of the host plant infection (i.e., tissue maceration) and hypersensitive response in nonhost tobacco plants. Therefore, the use of QS quenching strategies for biological control in E. carotovora subsp. atroseptica cannot prevent initial infection and multiplication of this pathogen. PMID- 15553253 TI - [Community-acquired pneumonia]. PMID- 15553254 TI - [Urinary tract infections: infrequent risk factor]. PMID- 15553255 TI - Food-intake and nutrients pattern in Italian adult male subjects. AB - Energy is necessary for all physiological functions of the body. Most of the vitamins are enzymes that require additional mineral as cofactors. Aim of the study was to evaluate the food intake and nutrients pattern in an adult population. 620 subjects underwent a complete medical visit, and filling the EPIC alimentary questionnaire. Comparison between nutrients intake and level assumption recommended nutrients (LARN) was assessed using one way analysis of variance (ANOVA) with Tukey-Kramer multiple comparison test. The daily medium caloric intake was 1837 +/- 654 Kcal. The daily mean iron assumption was 15.72 mg/die, statistically significant compared to 10 mg/die (LARN value) (p < 0.001). The calcium and zinc assumption was respectively 908.36 mg/die and 12.73 mg/die statistically different compared to 800 mg/die and 10 mg/die (LARN values) (p < 0.01). The assumption of the niacin was 20.24 mg/die significant different to 1.8 mg/die (LARN value) (p < 0.001). The assumption of vitamin A and C was respectively 1012.6 mcg/die and 142.92 mg/die significant different compared to 700 mcg/die and 60 mg/die (LARN values) (p < 0.01). The alimentary habits of the examined subjects did not present significant disequilibrium and evidenced a regular and good intake of micronutrients with anti-oxidative and anti-neoplastic activity. PMID- 15553256 TI - [Circadian rhythm of the renin-angiotensin-aldosterone system: a summary of our research studies]. AB - The renin-angiotensin-aldosterone system (RAAS) is characterized by a circadian rhythm (CR) whose acrophase is detectable early in the morning. The prorenin and angiotensin converting enzyme (ACE) show a CR as well. However, while the prorenin is in phase with the RAAS the ACE shows its circadian acrophase in the afternoon suggesting a negative feed-back. The RAAS CR is influenced by many factors. Its mesor is modified by sodium intake. The physical activity and rest affect both the mesor and acrophase. The variations in mesor amplitude and acrophase in aged subjects are conditioned by sex and physical activity. Moreover, the RAAS CR seems to be influenced by the race. In addition, it is abolished by the beta-adrenergic blockade, suggesting the existence of an adrenergic clock. Interestingly, the RAAS CR seems not to be a pacemaker for the blood pressure CR, whose acrophase is early in the afternoon. The RAAS CR is not substantially modified has in essential hypertension. However, the CR of plasma renin activity is disappeared in the low-renin essential hypertension, while the CR of plasma aldosterone is detectable. On the contrary, the aldosterone CR is not detectable in ascitic liver cirrhosis; but, it is restored when the ascites is removed by peritoneal-jugular shunt. No significant variation of the RAAS CR seems to occur in obesity and Cushing's syndrome. The RAAS CR has disappeared in Conn's disease as well as in Bartter's syndrome and Liddle's syndrome. The administration of indomethacine in Bartter's syndrome and of triamterene in Liddle's syndrome is able to restore the RAAS CR. Finally, the RAAS CR is not detectable in the heart or kidney transplanted patients; such a phenomenon could be attributed to cyclosporine and corticosteroids administration and to the denervation of the transplanted organs. PMID- 15553257 TI - [Community acquired pneumonia of hospitalised patient in the ordinary ward]. AB - Despite the evolution of the respiratory diagnostic methods and of the microbiological techniques and despite the introduction of new and effective antimicrobial drugs, community-acquired pneumonia (CAP) still remains a common and potentially lethal disease, as it represents the sixth general cause of death and the first cause of death concerning the infective diseases. The prolonged survival of patients affected by chronic respiratory and non-respiratory disorders and the worrying problem of antibacterial resistance showed by several pathogen agents constitute the possible causes of the epidemiological importance of CAP which still carries on significantly affecting health costs. In this review the author describes the epidemiological, etiological, microbiological, diagnostic and therapeutic aspects of CAP which requires hospitalisation in a non intensive care setting. PMID- 15553258 TI - [High-dose chemotherapy for metastatic breast cancer]. AB - In ninety's breast cancer was first in Europe for the use of high-dose chemotherapy with autologous hematopoietic stem cell transplantation in solid tumors in adults. Some phase II trials of high-dose chemotherapy showed high response rates and prolonged progression free survival in selected metastastic breast cancer patients. Few large, powerful randomized phase III studies comparing this approach with conventional chemotherapy have been completed: some studies showed a better progression free survival in favor of high dose chemotherapy, but no statistically significant difference in overall survival was observed. Many variables inside high dose chemotherapy program need to be considered. The identification of subsets of breast cancer patients who can benefit from high-dose chemotherapy is essential: high-dose chemotherapy should be included in a global treatment strategy, evaluating the integration with innovative treatment modalities, with the aim of eradicating the minimal residual disease in breast cancer patients achieving complete response after high dose chemotherapy. PMID- 15553259 TI - [54 old woman with recurrent urinary tract infection]. PMID- 15553260 TI - [The Moschkowitz syndrome in emergency department. Case report]. AB - Thrombotic thrombocytopenic purpura (TTP) is a life-threatening multisystem disorder of unknown etiology, first described by Moschkowitz in 1924. TTP is a thrombotic microangiopathy characterized by microvascular lesions with platelet aggregation. TTP is more common in adults and is associated with pregnancy; diseases such as HIV, cancer, bacterial infection, and vasculitis; bone marrow transplantation; and drugs TTP is a hematologic emergency. It is a multisystem disease that can cause rapid deterioration of the patient's neurologic, renal, and hematologic status. TTP is an uncommon disease with a high fatality rate if untreated or misdiagnosed. Rapid diagnosis and aggressive treatment by therapeutic plasma exchange are necessary to reduce the risk of a fatal outcome. Current clinical criteria for initiating therapy are: thrombocytopenia, and absence of other disease entities that could explain the thrombocytopenia. Early recognition and management are essential for patient survival. TTP is difficult to diagnose because the patient's presentation can be nonspecific and the characteristic pentad of symptoms may not occur together. Other disease entities can have some of the same symptoms. We discuss a case report of Moschkowitz syndrome in Emergency Department. PMID- 15553261 TI - [The oncology's rehabilitation ethical and care aspect]. PMID- 15553262 TI - [The reciprocal evaluation in the Learning Organization]. AB - INTRODUCTION: The culture of the appraisal was introduced in the Italian Universities only five years ago and the collected data come from the evaluation nuclei in the university on the basis of grills structured, proposed from the national Observatory for the Appraisal, where data are dealt in comparative key. MATERIALS AND METHODS: The Medicine faculty of the University Campus Bio Medico every year carries out an evaluation survey where the judgments from the students are compared with their didactic results. We utilise questionnaires to both inquire into the quality of the logistic and organizational aspects and into the didactic teacher competences and tutors, taking them in consideration singularly and as a whole, through the integrated course's analysis. RESULTS: This study represents one further development regarding those already published in the previous years, because it compares every teacher's results of the last the five years and every student's academic career from the first moment the student entered into the faculty. The two evaluative lines have been discussed in meetings with students, teacher and administrative staff. CONCLUSIONS: From the quantitative type of data gained through questionnaires, through the curves of performance of the students and through the interpretation that are in question observations for the improvement of the didactic quality are obtained. PMID- 15553263 TI - [Diet and management of degenerative diseases of the retina (retinitis pigmentosa)]. AB - Considerable progress has been made in the understanding and management of degenerative diseases of the retina. The dietetic intervention has been favourably proposed in the most common forms of retinitis pigmentosa, a condition potentially leading to blindness. Vitamin A has been shown to be effective in delaying progression of the disease. In these patients such treatment is the only possible therapy, to date, and a lifetime generous supplementation of retinol is advisable, together with a vitamin A-rich diet and/or a dietary supplement (e.g. carrot flour) or pharmacologic supplement of vitamin A. Supply of vitamin A in doses up to 25000 IU (7500 igr/day), even for several years, has so far proved safe from risk of occurrence of liver disease. A possible effect on hypercholesterolemia related to a very prolonged treatment in predisposed individuals can be avoided by using a special diet, particularly enriched with beta-carotene. Guidelines for preparing a diet, specially formulated to provide an elevated weekly supply of vitamin and/or its precursor (equal to 15000 IU or 5000 microg of RE, retinol equivalent) and to control possible risk factors related to dietetic manipulation (supply of fat lower than 30% of total calories, variable levels of cholesterol and polyunsaturated fatty acids n-3, n-6) are presented. As long as resolutive therapy is lacking, dietetic intervention plays a primary role, although underestimated, in the management of the patients suffering from retinitis pigmentosa. The diet is specifically characterized by presence of food with a high content of carotenoids, substances with a favourable and additive effect. PMID- 15553264 TI - [Acute diarrhea]. PMID- 15553265 TI - Adolescent archaics or adult moderns? Le Moustier 1 as a model for estimating the age at death of fragmentary supraorbital fossils in the modern human origins debate. AB - This study documents and examines selected implications of the adolescent supraorbital anatomy of the Le Moustier 1 Neandertal. Le Moustier's supraorbital morphology conforms to that expected of an adolescent Neandertal but indicates that significant development of the adult Neandertal torus occurs late in ontogeny. As the best preserved adolescent from the Late Pleistocene, Le Moustier 1's anatomy is used to help distinguish adolescent from adult anatomy in two cases of fragmentary supraorbital fossils, the Vindija late Neandertals and KRM 16425 from Klasies River Mouth (South Africa). It has been suggested that the modern-like aspects of the Vindija and Klasies supraorbital fossils are a function of developmental age rather than evolution. Although Le Moustier 1's anatomy does indicate that two of the Vindija fossils are adolescent; these two fossils have already been excluded from studies that demonstrate transitional aspects of the Vindija adult supraorbitals. Results of an analysis of KRM 16425 in light of Le Moustier 1 are more ambiguous. KRM 16425 is clearly not a Neandertal, but its morphology suggests that it may be an adolescent form of such late archaic Africans like Florisbad or Ngaloba. Both the Vindija and Klasies River Mouth cases highlight the need to be wary of confusing adolescent anatomy with modernity. PMID- 15553266 TI - Hominins are a single lineage: brain and body size variability does not reflect postulated taxonomic diversity of hominins. AB - Fossil hominin taxonomy is still debated, chiefly due to the fragmentary nature of fossils and the use of qualitative (subjective) morphological traits. A quantitative analysis of a complete database of hominin cranial capacities (CC, n = 207) and body weight estimates (Wt, n = 285), covering a period from 5.1 ma (millions of years) to 10 ka (thousands of years) shows no discontinuities through time or geographic latitude. Distributions of residuals of CC and Wt around regressions on date and latitude are continuous and do not differ significantly from normal. Thus, with respect to these characteristics, all hominins appear to be a single gradually evolving lineage. PMID- 15553267 TI - Association of frontal sinus development with somatic and skeletal maturation in Aboriginal Australians: a longitudinal study. AB - There have been very few studies of frontal sinus development and its association with other growth parameters in humans. The aim of this study was to investigate the relationship of frontal sinus development with somatic and skeletal maturation in a sample of Aboriginal Australians. The sample comprised 31 individuals, of whom 17 were males and 14 females. For the selected subjects the following records were available: lateral head radiographs, hand-wrist radiographs and stature recordings, generally covering the age range from 7 to 18 years. Descriptive statistics for frontal sinus size, skeletal ossification and body height were calculated at yearly intervals and comparisons made between the sexes. Growth velocities in frontal sinus height, frontal sinus depth and stature were also calculated for both sexes. The frontal sinus was found to display a well-defined adolescent growth spurt, with its peak velocity occurring after the peak velocity in body height. Females were found to attain peak velocity in sinus height earlier, on average, than males but they attained peak velocity in sinus depth at a similar age to males. The sequence of hand-wrist ossification events followed a similar pattern in both sexes, with events in females occurring approximately one year earlier than those in males. These results indicate that an adolescent spurt is present in frontal sinus growth and that the spurt tends to occur after statural velocity has peaked. Hand-wrist ossification events also tend to have a close relationship to peak statural and frontal sinus velocity and these relationships may be useful in clinical situations for predictive purposes. PMID- 15553268 TI - Sexual dimorphism in crown units of mandibular deciduous and permanent molars in Australian Aborigines. AB - Sexual differences in the crown units of mandibular molars were investigated in Australian Aborigines. The first and second deciduous molars (dm1 and dm2), and first to third permanent molars (M1, M2 and M3) were measured on dental casts using a sliding caliper. Measurements of tooth crowns included overall mesiodistal and buccolingual diameters, as well as the mesiodistal and buccolingual diameters of the trigonid and talonid. Percentage dimorphism values were greater in the talonid dimensions than the trigonid, indicating that sex differences tend to be larger in the later-developing crown units. Sex differences in mesiodistal diameters increased from dm1 to M2 but decreased for M3, the tooth that showed the least dimorphism of all the molars. This result seems to be due to the marked variability in size of the M3 between individuals. PMID- 15553269 TI - Who was the Red Queen? Identity of the female Maya dignitary from the sarcophagus tomb of Temple XIII, Palenque, Mexico. AB - The present investigation aims at contributing to the ongoing discussion on the unconfirmed identity of the Red Queen, a Classic Maya dignitary discovered in Temple XIII at Palenque, Mexico, by comparing her reconstructed facial profile to the portraiture of known female personages from the site. The comparison rests upon individual cranial features, like buccal prognatism, nasal root and inclination, chin prominence and the artificially shaped forehead. The similarities between the reconstruction, the female's funerary mask and local portraiture appear to identify the Red Queen as Lady Ix Tz'akb'u Ajaw (Ahpo Hel), the wife of Janaab' Pakal, one of the famous Maya rulers of the Classic Period. The proposed match and her family relationship with the king might explain the spatial closeness of their burial places in the Temple of the Inscriptions and Temple XIII. PMID- 15553270 TI - The facial restoration of Antal Simon, a Hungarian priest-teacher of the 19th c. AB - A large series of well documented, naturally mummified individuals were discovered during reconstruction works at the Dominican church in Vac in 1994- 1995. Among them was Antal Simon (1772--1808), a well-known priest and teacher, the director of the Institute of the Deaf in Vac in the 19th century. His remains were used to reconstruct the facial appearance, employing standard techniques. There is an authentic painted portrait of him. The finished facial reconstruction was compared to this portrait. As a measure of the effectiveness of the reconstruction, the technique of superimposition was also used, which represented a direct comparison of the features of the portrait with those of the reconstructed face. The topographical, morphological and anatomical characteristics of the two faces were compared with the following results: 62% of the reconstructed traits showed great resemblance to the original face, 35% showed close resemblance and 3% showed approximate resemblance. PMID- 15553271 TI - Hyperostosis frontalis interna: archaeological evidence of possible microevolution of human sex steroids? AB - Hyperostosis frontalis interna is a restricted bilateral thickening of the frontal endocranial surface, which is frequently found in postmenopausal females today. Surprisingly, this condition had a higher male prevalence in its rare archaeological records. This is again highlighted by the oldest known male European hyperostosis frontalis interna case in an adult Celtic from 100 BC presented here. This unique specimen supports earlier suggestions of the possible microevolution of human endocrine regulation, e.g. by sex steroids, and its pathoanatomical impact. PMID- 15553272 TI - Palatine torus in the pre-conquest inhabitants of the Canary Islands. AB - An unusually high frequency of palatine torus in prehistoric Canary Islands skeletal remains is investigated in terms of population origins, evolutionary forces (gene flow and genetic drift) and environmental effects. Palatine torus frequencies (percent presence) are compared between the Canary Islands sample and skeletal samples from proposed ancestral regions, including northwest Africa, northeast Africa, sub-Saharan Africa, southern Europe, northern Europe and western Asia. The frequency of palatine torus is much higher in the Canary Islands sample (23.8%) than in samples from Northern and sub-Saharan Africa (1.8 6.1%), southern Europe (7.5%) and western Asia (2.1%), but is much lower than in the sample from northern Europe (57.4%). Because biological and archaeological evidence provide strong support for a northwest African origin for indigenous Canary Islands populations, the relatively high occurrence of palatine torus in the Canary Islands populace cannot be explained by an alternative ancestry; rather, it may be best explained by either evolutionary forces such as genetic drift or gene flow, or by environmental forces, such as hard chewing or a heavy dependence on marine foods. Genetic drift and gene flow seem less likely, since frequencies of other dental traits known to be under strong genetic control do not differ greatly between the prehistoric Canary Islands and northwest African samples. Environmental factors such as diet seem the most likely explanation and may include heavy consumption of marine foods, which has been implicated in torus formation. PMID- 15553273 TI - A comparison of inbreeding rates in India, Japan, Europe and China. AB - This is a report on an application of a new method for estimating inbreeding rates in large, semi-isolated populations over time. This study is intended to further show the versatility of the method. In a previous study, the method was shown to be useful in analysing the effect of details in a single population curve on inbreeding. In this study, the method is applied to the population curves of India, Japan, Europe and China over the past two millennia, to allow inter-comparisons of the inbreeding within those populations. Anthropologists traditionally concentrate on small isolates within a country for the estimation of inbreeding. Those isolates, however, might not be representative of the country as a whole. The method used in this study attempts to estimate inbreeding over large regions over an extended period of time. The method models the genealogical 'paradox' and yields estimates of the average inbreeding in terms of Pearl's coefficient Z, as a function of time. It is first assumed that the whole population of each country is the adult (breeding) population, corresponding to minimum inbreeding. It is found that the more complex the population curve examined, the greater the precision of the Z curves obtained. The effect of incorporating a single known value of inbreeding into the analyses is also investigated. This procedure produces a more realistic situation where the adult (breeding) population is considerably less than the entire population. It is shown, that if it is assumed that the whole population of the country is the adult (breeding) population, then a present day person from one of the four regions/countries studied is descended from between 72% and 97% of the early medieval population of the particular country. On the other hand, if a known value of inbreeding for Britain is incorporated into the analysis, these values become of the order of 1% for the older-settled regions/countries, and about 16% for Japan. However, that value for Japan is reduced to about 1% when a known value of inbreeding for this country is used. Although some uncertainty in these results remains, the versatility of the method demonstrated here will provide more accurate results, as better input-data become available. PMID- 15553274 TI - The maturation rate of girls living in rich and poor rural regions of Poland before and after the transformation of 1989. AB - The aim of this study was to find out whether the political and economic system transformation in Poland in 1989 had an influence on the rate of maturation of girls from various categories of the rural population in two regions, differing in wealth. The stratification of the villagers was based on the source of the family income (farmers, farmer-workers and non-farmers), on parents' education, and on the number of children per family. The age at menarche (AM) was used as biological indicator of living conditions. Rural girls aged 9.5-18.5 years were studied in 1987 in Leszno (L), a rich region (n = 2049), and in 1989 in Suwalki (S), a poor region (n = 2077). The study was repeated in 2001 in the same villages, the numbers of subjects amounting to 2440 and 2122, respectively. In the 1980s, AM in regions L and S amounted to 13.18 and 13.88 years, respectively, and was strongly related to social stratification. The earliest AM was found in daughters of non-farmers and the latest in the girls from farmers' families. In 2001, the acceleration in AM amounted to 0.14 years in the rich region and to 0.46 years in the poor region. The percentage of families owning a car, freezer, colour TV and automatic washing machine markedly increased and the education of parents improved. Such results are indicative of an improvement in the living standard of the groups investigated, but the villagers regard themselves as the losers as a result of the system transformation in Poland. Since the Polish population is considered as ethnically homogenous, the great differences in AM within the rural population, as well as the differences between the inhabitants of villages and large cities should be regarded as biological effects of social inequalities. Such differences indicate that Polish rural girls are far below the genetic potential for menarcheal age. PMID- 15553275 TI - The tiresome second best. PMID- 15553276 TI - The flu and you. PMID- 15553277 TI - Bones, beautiful bones. PMID- 15553278 TI - Curing campus blues. PMID- 15553279 TI - [Medicolegal aspects of witnessed suicide due to gunshot to the head. Part 1: Circumstances and psychopathology]. AB - In the USA the frequency of witnessed suicide, i.e. suicide committed in the presence of at least one further individual, is reported to range between 5 and 15%. As up to now no detailed analysis of this special issue has been given in the medicolegal literature, this article presents and discusses a number of cases including 8 males and 1 female (age 19-58 years, mean 41.2 years) who had all committed suicide by inflicting a gunshot to the head. In 3 cases post-mortem blood alcohol concentrations of 1.73, 1.88 and 2.23 g/kg respectively were observed. Toxicological tests produced negative results. 5 cases had a medical history of psychiatric disorder with endogenous depression in 3, chronic alcohol abuse and drug abuse with concomitant psychosis in one case each. As far as the motives were known, domestic arguments were of prevailing importance. With one exception the suicides were committed at home. In 6 cases one witness was present, in 2 cases 2 individuals and in one case 22 persons witnessed the suicide. The reported cases are discussed in comparison with the psychiatric psychological classification according to McDowell et al. with the aim to provide a solid and comprehensive medicolegal method to distinguish between homicide and suicide. Furthermore aspects of psychiatric sequelae and psychological support of the witnesses, which are also of importance for the forensic pathologist called to the scene, are discussed. PMID- 15553280 TI - [Death by a cleaning pig--an unusual accident at work]. AB - A worker was killed operating on a natural gas pipeline. At a receiver station the man intended to pick up two so called cleaning pigs, each weighing about 150 kg, diameter 0.5 m. For this purpose the gas pressure in the pig trap was let off by a valve, manometer controlled. According to an eyewitness's statement, the receiver pressure was equal to outside air pressure before the accident. The victim stood right in front of the flap of the receiver when he began to unfix the screws of the flap. Whilst working, the flap snapped out driven by the two cleaning pigs. The man and the devices were flung through a wire-netting fence and dropped down on a nearby field at a distance of 27, 29 and 38 m, respectively, from the receiver. The man died on the scene of the accident. The forensic autopsy ordered by the prosecution revealed signs of massive blunt trauma on the head, thorax and abdomen. In the criminological and forensic reconstruction of the accident the external injuries of the victim were found to be consistent with the front surface of the cleaning pig flung out first. It was determined that the second pig had got stuck in the receiver and that gas pressure had built up behind the pigs due to a leaky valve. As a consequence the pigs were expelled at a velocity of approximately 220 km/h causing a pattern of injury comparable to that of a fall from a great height. PMID- 15553281 TI - [Decapitation due to early post-mortem canine gnawing]. AB - A 53-year-old male was found dead in his home. The deceased's head, almost totally skeletonized, was lying at a distance of about 150 cm from the thorax inlet. The other occupant of the flat was a mongrel sheepdog. The autopsy conducted for the inquest established extensive damage to soft tissue in the head, neck and chest. The second to sixth cervical vertebrae were missing. The seventh cervical vertebra and the right first rib displayed bone lesions. The tissue injuries were attributed in the diagnosis to post-mortem canine gnawing. Cause of death was given as intermittent haemorrhaging of the gastro-intestinal tract from oesophageal varices in a status of hepatic cirrhosis. There was no indication that death had been caused by another party. About two days were estimated to have elapsed since death. Attention is drawn to this doubtless rare instance of total decapitation of the deceased with displacement of the head caused by a dog during the early post-mortem period. PMID- 15553282 TI - [Suicidal monointoxication with flunitrazepam. Further comment on coloration phenomena of the upper gastrointestinal tract]. AB - Fatal monointoxications with benzodiazepines, for instance with a suicidal intention, are exceptional findings. We report the autopsy case of an 82-year-old woman who died due to a suicidal monointoxication with Rohypnol 1 mg film tablets (therapeutical agent: flunitrazepam). 0.065 mg/L flunitrazepam and 0.34 mg/L 7 aminoflunitrazepam were detected in a postmortem heart blood sample and toxicological analysis revealed the metabolite 7-aminoflunitrazepam in gastric contents, too. At external examination, a bluish-turquoise coloration was seen around the woman's right nostril and within the oral cavity. At autopsy, similar coloration phenomena were seen on the mucosa of the distal esophagus and the stomach. Formerly, bluish stains on mouth and nostrils were considered indicative of intoxications with organophosphates such as parathion (E 605). More recently, case reports accumulate where an intoxication with Rohypnol 1 film tablets (containing the coloring agent indigocarmine in its core) have to be considered as a potential differential diagnosis of such coloration phenomena. PMID- 15553283 TI - [About the so-called "Geserick sign"]. AB - Geserick et al. were the first who reported about fractures of the medial and basal wall of the orbita, originating from blunt occipital trauma leading to consecutive contrecoup lesions of the bulbus/orbita complex. A case history with (survived) occipital craniocerebral trauma and monocle hematoma (48-year-old drunken homeless person, found unconscious in a small street of the redlight quarter) is presented. The question was whether the injuries were caused by falling or inflicted by an assailant. PMID- 15553284 TI - [Radiologically determined DMF index variations for forensic age estimation of young adults]. AB - The eruption and mineralization of third molars are the main criteria for dental age estimation of living people involved in criminal proceedings. Since the development of third molars is usually completed by the age of 19 or 20 years, this feature cannot be used to ascertain whether a person has attained the forensically relevant age of 21 years. This study examined whether the DMFT index or the third molar DFT index could supply reliable information enabling conclusions to be drawn about whether a person has reached the age of 21. To this end, 650 conventional orthopantomograms of German subjects aged 18 to 30 years were evaluated. The DMF index variations considered were found to be unsuitable for determining whether a person is over 21 because they displayed a weak correlation with age and considerable individual scatter. A high indicative value can only be attributed to a third molar DFT index of 4, although this is rarely observed in the relevant age group. An X-ray examination of the medial clavicular epiphyseal cartilage is, therefore, strongly recommended when assessing whether a person has attained 21 years of age. PMID- 15553285 TI - The effect of adhesive and flowable composite on postoperative sensitivity: 2 week results. AB - OBJECTIVE: To measure 2-week postoperative sensitivity in Class II composite restorations placed with a self-etching adhesive (Clearfil SE Bond) or a total etch adhesive (Prime&Bond NT) with or without a flowable composite as cervical increment. METHOD AND MATERIALS: Upon approval by the University of Guarulhos Committee on Human Subjects, 100 restorations were inserted in 46 patients who required Class II restorations in their molars and premolars. Enamel and dentin walls were conditioned with a self-etching primer (for Clearfil SE Bond) or etched with 34% phosphoric acid (for Prime&Bond NT). A 1- to 2-mm-thick increment of a flowable composite (Filtek Flow) was used in the proximal box in 50% of the restorations of each adhesive. Preparations were restored with a packable composite (Surefil). The restorations were evaluated preoperatively and 2 weeks postoperatively for sensitivity to cold, air, and masticatory forces using a visual analog scale. Marginal integrity of the accessible margins was also evaluated. Statistical analysis used a mixed linear model with subject as a random effect. RESULTS: Ninety-eight teeth from 44 subjects were observed at 2 weeks. The type of adhesive and use of flowable composite had no significant effects or interaction for any of the four outcomes of interest, ie, change from baseline to 2 weeks in sensitivity and response time for the cold or air stimulus. For the air stimulus, the overall average change from baseline was not significant for either sensitivity or response time. For the cold stimulus, the overall average change from baseline was significant for both sensitivity and response time. No case of sensitivity to masticatory forces was observed. CONCLUSION: No differences in postoperative sensitivity were observed between a self-etch adhesive and a total-etch adhesive at 2 weeks. The use of flowable composite did not decrease postoperative sensitivity. PMID- 15553286 TI - Clinical evaluation of proximal contacts of Class II esthetic direct restorations. AB - OBJECTIVE: This study evaluated the effectiveness of proximal contacts in Class II restorations using two types of matrix bands (steel and polyester) with two different restoration techniques (incremental and with prepolymerized particles). METHOD AND MATERIALS: Eighty-eight Class II adjacent restorations using Prodigy resin composite were performed: 44 with the incremental technique (22 with steel matrix bands, 22 with polyester matrix bands) and 44 utilizing prepolymerized resin particles (22 steel matrix, 22 polyester matrix). The restorations were clinically evaluated at baseline and at 6, 12, and 18 months. Proximal contacts obtained immediately after restoration procedure in all restorations were satisfactory. RESULTS: No statistically significant alterations were found in 18 months of evaluation. CONCLUSION: Regardless of the utilized resin composite, there were no differences in the amount of proximal contact variations with respect to tested techniques and matrices. PMID- 15553287 TI - Microleakage of different resin composite types. AB - OBJECTIVE: The purpose of this study was to evaluate the microleakage of Class V cavity preparations restored with three different types of resin composite and an ormocer. METHOD AND MATERIALS: Forty Class V cavities were prepared in buccal and lingual surfaces of 20 recently extracted molar teeth. The occlusal margin of each restoration was on enamel and the gingival margin on dentin. Teeth were randomly assigned to four groups of 5 teeth each and restored as follows: Group I, flowable resin composite (Tetric Flow); Group II, hybrid resin composite (Z100); Group III, packable resin composite (Solitaire 2); Group IV, organically modified ceramics-ormocer (Admira). In all groups, the manufacturers' instructions were strictly followed. All restorative resin composite materials were placed in one increment. All teeth were then immersed in 0.5% basic fuchsin solution for 24 hours after thermocycling (200 cycles; between 4 degrees C to 60 degrees C). The teeth then were longitudinally sectioned and observed under a stereomicroscope. The degree of dye penetration was recorded and analyzed with the Kruskal-Wallis and Mann-Whitney tests. RESULTS: No statistically significant differences in microleakage were observed between groups either on enamel or dentin. CONCLUSION: All restorative materials demonstrated equal effectiveness in reducing microleakage. PMID- 15553288 TI - In vitro study of surface microhardness of a resin composite exposed to a quartz halogen lamp. AB - OBJECTIVES: To determine the minimum exposure times consistent with optimized surface microhardness numbers (VHN) for the top and bottom surfaces, respectively, of a resin composite (1.5-mm depth) cured using a quartz-halogen activation lamp. Additionally, to determine the conditions appropriate to achieve optimized bottom/top hardness ratios. METHOD AND MATERIALS: Disk specimens of a commercial resin composite restorative (Spectrum TPH) were exposed to a quartz halogen lamp, and VHNs were measured for the top and bottom surfaces at elapsed times of 1 hour and 24 hours. RESULTS: Data were dependent on exposure time, time elapsed after exposure, and the surface involved. Exposure time featured limiting values in excess of which no significant increase in microhardness took place. All microhardness data increased significantly over an elapsed time interval between 1 hour and 24 hours postexposure. Microhardness ratios (bottom/top) were dependent on exposure time only and were not influenced by time elapsed after exposure. CONCLUSION: Minimum exposure time may be considered as the greater of the times to achieve optimized top hardness and optimized hardness ratio, respectively. PMID- 15553289 TI - In vitro influence of Carisolv on shear bond strength of dentin bonding agents. AB - OBJECTIVE: The aim of this study was to evaluate the influence of Carisolv on the shear bond strength (SBS) of hydrophilic adhesive systems in dentin. METHOD AND MATERIALS: Two hundred bovine incisors were polished with 320- to 1,000-grit aluminum oxide paper and randomly divided into 10 groups (n = 20). The adhesive systems were used according to manufacturer's instructions, and a restorative resin composite (Definite) was inserted in a Teflon matrix and cured. The specimens were kept in humid storage for 7 days at 37 degrees C. The SBS tests were performed in an EMIC universal testing machine with a crosshead speed at 0.5 mm/min. The mean values (MPa +/- [SD]) were analyzed with analysis of variance and Dunnett and Tukey tests. RESULTS: The results were expressed in decreasing values (MPa): G6 = 18.55 +/- 6.95; G3 = 17.55 +/- 5.73; G2 = 17.05 +/- 5.33; G8 = 16.95 +/- 7.13; G7 = 16.94 +/- 5.22; G4 = 16.31 +/- 3.38; G1 = 15.85 +/- 4.45; G9 = 14.21 +/- 5.87; G5 = 12.89 +/- 3.79; and G10 = 12.31 +/- 5.18. CONCLUSION: Carisolv did not interfere in the adhesion to dentin, but its cleanser technique did interfere. PMID- 15553290 TI - Cemento-ossifying fibroma: a case report. AB - Cemento-ossifying fibroma is a mesodermal, slow-growing, benign fibro-osseous lesion of the jaws. A case of mandibular molar enlargement that was treated previously by other surgeons and diagnosed as cemento-ossifying fibroma, is presented. Two recurrences occurred 4 and 8 years after surgery due to incomplete surgical removal. PMID- 15553291 TI - Pre- and posttreatment analysis of clinical symptoms of patients with temporomandibular disorders. AB - OBJECTIVE: This study treated patients with temporomandibular disc displacement with reduction (with pain, limited mandibular movement, and clicking sound symptoms) using a combination of analgesics, injection, mandibular exercise, and occlusal splints. METHOD AND MATERIALS: Twenty-five patients with temporomandibular joint (TMJ) disorders were evaluated for pretreatment complaints and clinical findings, such as TMJ sounds, mandibular deviation, limited mouth opening, and bilateral magnetic resonance imaging results. Diagnostic treatment was then planned; all patients received occlusal splints, and 10 patients received injections. RESULTS: Evaluations were conducted 1 year after the initial diagnosis and treatment. Complaints (especially of pain) by the patients who received injections had reduced significantly. After 6 months of occlusal splinting, clinical findings of patients with TMJ disc displacement had greatly improved. To manage parafunctional habits of the patients, night plate usage was continued. Therefore, clinical symptom reduction was maintained. CONCLUSION: The use of mandibular manipulation technique can decrease the anterior disc displacement of the TMJ. PMID- 15553292 TI - The prevalence of inflammatory and developmental odontogenic cysts in a Jordanian population: a clinicopathologic study. AB - OBJECTIVE: To determine the prevalence of odontogenic jaw cysts in a Jordanian population and to compare these data with previously published reports from other geographic areas. METHOD AND MATERIALS: The files on odontogenic jaw cysts treated between 1989 and 2001 in the Oral and Maxillofacial Pathology Diagnosis Service at the Department of Oral Medicine and Oral Surgery, Faculty of Dentistry, Jordan University of Science and Technology, were reviewed. Clinical and radiographic data were recorded and microscopic slides evaluated according to the most recent World Health Organization classification. Cases were analyzed with regard to age, sex, and anatomic site. RESULTS: A diagnosis of odontogenic jaw cyst was established in 654 patients, with a male-to-female ratio of 1.7:1. Radicular cyst was the most common type of odontogenic cyst found (41.7%), followed by dentigerous cysts (24.8%). The peak age affected was between the third and fifth decades. Both jaws were almost equally affected. The most common anatomic site of incidence was the maxillary incisor/canine region, followed by the mandibular molar region. CONCLUSION: This study indicates that there are some geographic differences with regard to the relative frequency, sex, and anatomic distributions of odontogenic cysts. PMID- 15553293 TI - Dimensions of metal framework components of metal-ceramic fixed partial dentures constructed in a dental school setting. AB - OBJECTIVES: The aims of this study were to measure the dimensions of connectors and retainer copings of metal-ceramic fixed partial denture (FPD) frameworks and to explore, by means of a questionnaire, the factors viewed by technicians as important in their planning and provision of such dimensions. METHOD AND MATERIALS: Using two calliper devices, measurements were made of 66 consecutive FPDs under production in a dental school-based laboratory. The dimensions of 176 connectors and 124 retainers were analyzed with reference to FPD length and to number of pontics in a span. Questionnaires were completed by all 14 technicians working in the fixed prosthodontics section of the laboratory. RESULTS: The largest mean vertical connector dimensions were in the anterior regions of both arches, while the largest mean horizontal dimensions were in the posterior regions. Dimensions were unrelated to FPD size and length of interabutment span. The minimum thickness of the retainer coping walls was generally below optimal recommendations. Available space was the factor most commonly cited by technicians as dictating the dimensions that they applied. CONCLUSION: Dimensions were generally smaller than current recommendations, which the questionnaire responses indicate are largely due to space constraints. PMID- 15553294 TI - Effects of different disinfectants on physical properties of four temporary soft denture-liner materials. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of some commonly used disinfectants on physical properties of four temporary soft denture-liner materials. METHOD AND MATERIALS: Soft-liner materials, including three plasticized acrylic resin-liners (Tempo, Immediate, Flexacryl Soft) and one silicone-based material (Ufi Gel P) were studied. Specimens prepared with the soft-liner materials were tested after the application of disinfectants (5% Deconex and 3.5% Savlex for 10 minutes; 2% sodium hypochlorite and 5.25% sodium hypochlorite for 5 minutes) and were compared with the control group (not disinfected; immersed in distilled water). Five specimens of each material were tested and data collected on days 4, 7, 11, and 15. The hardness values were determined using the Shore A hardness test. RESULTS: Water sorption test results showed that the disinfectants had statistically significant effects on water sorption values (except days 7 and 15 for Flexacryl Soft and days 4, 7, and 11 for Ufi Gel P). Water solubility and hardness test results also showed statistically significant effects on the materials by the disinfectants. CONCLUSION: Disinfectants significantly affected the physical properties of water sorption, water solubility, and hardness of the soft-liner materials. PMID- 15553295 TI - Secondary trauma from occlusion: three-dimensional analysis using the finite element method. AB - Clinical effects of forces applied by dental occlusion on the periodontium have been evaluated for decades. Historically, trauma from occlusion has been considered as a major etiologic factor of inflammatory periodontal diseases, while some researchers have interpreted it to be of less importance or without any detectable importance in periodontics. In this study, five three-dimensional models of a maxillary central incisor were created using ANSYS 5.40. The only difference in each model was the height of the alveolar bone that showed from normal height (13 mm of alveolar bone height) to 8 mm of alveolar bone loss (5 mm of alveolar bone height). Five-point forces of 0.3 N summing up to 1.5 N were applied in a parallel line, 1 mm apical to the incisal edge on the palatal side in a palatolabial direction. The maximum (S1) and minimum (S3) principal stresses in the nodes of the labial side of the periodontal ligament (apical to the alveolar crest) were assessed. Analysis was done using the finite element method. An increase of S1 (up to 16 times in the cervical and 11.25 times in the apical area) and S3 (up to 17.13 times in the cervical and 9.9 times in the apical area) in comparison to the normal model was shown. The highest stress levels were traced in the subcervical area, except for the last model (8 mm of the alveolar bone loss). According to the results of this study, 2.5 mm of alveolar bone loss can be considered as a limit beyond which stress alterations were accelerated. Based on the FEM analysis, alveolar bone loss increases stress (S1 and S3) produced in the PDL, in spite of applying the same force vector. PMID- 15553296 TI - In vitro comparison of dentin bonding systems: effect of testing method and operator. AB - OBJECTIVES: The purpose of this study was to investigate the influence of different in vitro tests on the ranking of four dentin bonding systems (OptiBond FL, Solid bond, OptiBond Solo, Etch&Prime 3.0) and to determine operator influence on push-out test results. METHOD AND MATERIALS: One-hundred eighty extracted human molars were used in this study. For all bonding systems, bond strengths were determined using the shear, tensile, and push-out tests. In the push-out test, two series of specimens were bonded by two operators. Additionally, margin adaptation in Class V cavities was evaluated using scanning electron microscopy and dye penetration. RESULTS: Both test methods and bonding systems revealed a significant influence on bond strength. Ranking of mean bond strengths was identical in the shear and tensile tests but different in the push out test. In addition, material ranking in the push-out test was found to be operator dependent. No correlation was observed between bond strength and margin adaptation. A weak correlation was found between the percentage of gap formation and dye penetration in Class V cavities. CONCLUSION: Comparisons of dentin bonding systems based on the results of different in vitro tests may be misleading. Operator influence on the results of push-out tests require further consideration. PMID- 15553297 TI - Assessment of aortic stenosis severity. PMID- 15553298 TI - Coenzyme Q10 in patients with end-stage heart failure awaiting cardiac transplantation: a randomized, placebo-controlled study. PMID- 15553299 TI - Chinese first name comes last. PMID- 15553300 TI - Not just another restenosis trial. PMID- 15553301 TI - Keeping the autopsy alive. AB - The recent decline in autopsy rates has many causes and has been the subject of vigorous debate amongst clinicians, pathologists, and investigators. For cardiologists, the lack of direct information on the vessel wall may impede the ability to develop and interpret new imaging methods and to understand the effects of the newest interventions. PMID- 15553302 TI - Women and the implantable cardioverter defibrillator: a lifespan perspective on key psychosocial issues. AB - The clinical success of the implantable cardioverter defibrillator (ICD) in reducing mortality suggests that more women will be receiving ICDs in the future. The impact of ICD therapy in women is unique in western societies; the ICDs scar and lump in the pectoral area can lead to body image concerns due to the emphasis on women's physical attractiveness. Social support and roles are challenged because women's reaction to stress has been characterized by a "tend and befriend" response, involving cultivating and utilizing social networks, rather than the "fight or flight" response more typical of men. In addition, a woman's identity as a caretaker and caregiver can be threatened by the actual and perceived activity limitations imposed by the ICD or the underlying heart condition. Finally, reproductive and sexual health are important issues, as 25 to 50% of patients with ICDs report concerns in this area, but also report discomfort in discussing these concerns with their health care providers. The purpose of the present paper is to review the relevant literature and to identify the unique impact of the psychosocial issues of body image, social support and roles, and sexual development and reproductive functioning for women with ICDs across the lifespan. In the absence of complete empirical research data on the impact of these concerns, hypotheses to test in future research are offered. PMID- 15553303 TI - Should standard medical therapy for angina include a statin? AB - Although a wealth of evidence supports the use of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) in patients with clinically evident coronary artery disease, these agents are still underutilized. Statins are the most effective agents in reducing low-density lipoprotein-cholesterol among lipid-lowering drugs, and studies have recently shown that they improve endothelial function and plaque stabilization, and induce regression of atherosclerotic lesions. This article reviews the most recent evidence and guideline recommendations supporting the use of statins in chronic stable angina pectoris and acute coronary syndromes. PMID- 15553304 TI - Images in cardiology. Congenitally corrected transposition of the great arteries with subpulmonic stenosis. PMID- 15553305 TI - Images in cardiology. Disappearance of cardiac gallium uptake after corticosteroid therapy for sarcoidosis. PMID- 15553306 TI - Gender as a risk factor for contrast nephropathy: effects of hydration and N acetylcysteine. AB - BACKGROUND: In a few studies, N-acetylcysteine has been shown to prevent contrast induced nephropathy in patients with chronic, stable renal failure undergoing elective procedures. Other studies have shown variable outcomes. Furthermore, the majority of prior studies have mainly studied men, and gender as a risk factor has not been studied. HYPOTHESIS: The study sought to evaluate the effectiveness of N-acetylcysteine and hydration in unselected patients with both acute and stable renal insufficiency (RI) undergoing urgent or elective cardiac or peripheral angiography. METHODS: We evaluated records of 146 patients with RI undergoing angiography. We compared patients receiving periprocedure hydration and acetylcysteine with patients who were only hydrated or received no pretreatment. We evaluated the 48-h change in serum creatinine between groups and further analyzed the effect of hydration and gender on outcomes. RESULTS: Demographics and baseline creatinine were similar between groups. Post procedure, the creatinine increased significantly in both groups, but less so in the acetylcysteine group (control: 0.35 +/- 0.08 mg/dl; acetylcysteine: 0.14 +/- 0.04 mg/dl, p < 0.05). When the control group was further stratified by hydration, the increase in creatinine for the hydrated patients was only 0.17 +/- 0.10 mg/dl compared with 0.54 +/- 0.12 mg/dl in patients with inadequate hydration. In the control group, women were more likely to receive no preprocedural hydration (59 vs. 40%), had a bigger rise in creatinine, received less protection from hydration alone, but were equally well protected by hydration plus acetylcysteine. In the acetylcysteine group, change in creatinine for women was minimal (+ 0.14 +/- 0.07 mg/dl) and not different from men (+ 0.15 +/- 0.05). CONCLUSION: Unselected patients with acute and chronic RI had no benefit from acetylcysteine beyond that seen with hydration alone. Gender may be a risk factor for contrast-induced nephropathy, with hydration offering less protection in women. Acetylcysteine (with hydration) seems to minimize the gender difference. PMID- 15553307 TI - Serum levels of cardiac enzymes before and after renal transplantation. AB - BACKGROUND: Cardiac troponins are very sensitive and specific indicators of myocardial damage; however, they are sometimes found to be increased in patients with end-stage renal disease (ESRD). HYPOTHESIS: The aim of this study was to determine cardiac troponin I (cTpI) and creatine kinase myocardial isoform (CK MB) levels and to assess their diagnostic and prognostic importance before and after renal transplantation. METHODS: Thirty-four patients with ESRD (mean age 31.8 +/- 8.6 years, 11 women) were enrolled. Serum levels of cTpI and CK-MB were measured pre- and postoperatively on Days 1, 7, and 30. Patients were followed up for cardiac events, and possible myocardial damage was investigated by exercise thallium-201-labeled myocardial perfusion scintigraphy. Mean cTpI levels were 0.24 +/- 0.11 ng/ml (preoperative), and 0.34 +/- 0.27 ng/ml (Day 1), 0.26 +/- 0.11 ng/ml (Day 7), and 0.28 +/- 0.30 ng/ml (Day 30). RESULTS: Compared with preoperative levels, cTpI was increased in 16 (47%), decreased in 6 (17.6%), and did not change in 12 (35.4%) patients. However, the increase did not exceed the myocardial infarction reference level of 2.3 ng/ml in any patient. Mean CK-MB levels were 12.6 (8.7U/l (preoperative), and 16.8 +/- 9.2U/l (Day 1), 16.3 +/- 8.1U/l (Day 7), and 13.3 +/- 6.6U/l (Day 30). Creatine kinase-MB was increased to above normal levels of 24 U/l in 13 (38.2%) patients on postoperative Days 1 or 7, and decreased to normal at the end of Month 1. No cardiac events occurred, and there was no abnormality in any patient on thallium scintigraphy. CONCLUSION: There was no significant difference in the levels of cTpI in patients with ESRD without cardiac events before and after renal transplantation (p > 0.05). Our findings show that cTpI has very high sensitivity and specificity for detecting cardiac damage in patients with ESRD after renal transplantation. PMID- 15553308 TI - A low-carbohydrate diet in overweight patients undergoing stable statin therapy raises high-density lipoprotein and lowers triglycerides substantially. AB - BACKGROUND: A low-carbohydrate diet remains controversial, especially in patients with arteriosclerotic heart disease. HYPOTHESIS: This study was undertaken to evaluate the effect of a low-carbohydrate diet on the lipid levels in obese patients with known arteriosclerotic heart disease on chronic statin therapy. METHODS: Thirty-eight overweight patients with angiographically documented arteriosclerotic heart disease were followed in a private cardiology practice setting. All patients were undergoing stable statin therapy. Patients received a 15-min consultation and a 4-page pamphlet explaining a low-carbohydrate diet; no other diet instruction was given. Patients were followed weekly for 2 weeks, then monthly for 3 months, then every third month. A fasting finger stick lipid panel (cholesterol, high-density and low-density lipoprotein [HDL/ LDL], triglycerides, and glucose) was obtained with each visit and patients were weighed in street clothes. RESULTS: The 38 patients were followed for a average of 11.8 months (range 6-22 months). Average body mass index declined from 33.5 kg/m2 before to 27.9 kg/m2 at the end of the study. Weight loss averaged 31 lbs (range 16-107 lbs). Triglyceride levels were lowered by 29.5%, HDL raised by 17.6%, and cholesterol decreased by 8.4%. The cholesterol/ HDL ratio changed from 5.31 to 3.78 and LDL cholesterol decreased by 5%. CONCLUSION: The addition of a low carbohydrate diet for overweight patients with known coronary artery disease undergoing stable statin therapy causes significant weight loss and a favorable change in the lipid panel. PMID- 15553309 TI - Aortic valve replacement in patients 70 years and older. AB - BACKGROUND: Aortic valvular disease is the most common valvular lesion among elderly patients. Because of changing demographics, it has become increasingly frequent. Aortic valve replacement (AVR) is the only effective treatment for aortic valvular disease. HYPOTHESIS: This study was undertaken to evaluate the results of AVR in an elderly population. METHODS: Data were retrospectively analyzed in 117 consecutive patients (mean age 73.8 years) who underwent AVR between 1991 and 2002. RESULTS: Pure or predominant severe aortic stenosis was present in 108 patients. Nine patients had severe aortic regurgitation. Before valve replacement, 62.4% of the patients were in New York Heart Association (NYHA) functional class III-IV. A bioprosthesis was implanted in 62.4% of the patients, and 37.6% received a mechanical valve. Concomitant cardiac surgical procedures were performed in 25 patients (coronary artery bypass graft in 22, mitral valve replacement in 3). There were 17 deaths, giving a perioperative mortality rate of 14.5%. Multivariate logistic regression showed that repeat surgery for bleeding, prolonged cardiopulmonary bypass time, postoperative respiratory failure, and postoperative acute renal insufficiency were significant independent predictors of operative mortality. Of the 100 hospital survivors, 78 were followed for a mean of 42.9 months. There were six deaths during follow-up; only two of these were cardiac related. Five-year actuarial survival for all patients and for hospital survivors were 70 and 91.1%, respectively. One year post surgery, all patients were in NYHA functional class I-II. CONCLUSION: In a selected patient population, AVR in the elderly is associated with acceptable mortality and morbidity. The outlook for hospital operative survivors is excellent with improved quality of life and an expected survival normal for this particular age. PMID- 15553310 TI - Characteristics of contemporary patients with hypertension and coronary artery disease. AB - BACKGROUND: Despite a high prevalence of hypertension in the population with CAD, there are limited data describing the clinical characteristics and treatments, as well as their interrelations in these patients. This is particularly true for black and Hispanic patients who have been underrepresented in randomized CAD trials. HYPOTHESIS: There exist racial and ethnic differences that define the characteristics of patients with both coronary artery disease (CAD) and hypertension. METHODS: This report describes the characteristics of Caucasian, Hispanic, and black patients enrolled in the International Verapamil SR/trandolapril Study (INVEST), a prospective trial undertaken exclusively in patients with CAD and hypertension. RESULTS: In all, 10,925 Caucasian, 8,045 Hispanic, and 3,029 black patients are described. An abnormal angiogram or documented myocardial infarction was observed more frequently in Caucasian patients (73%), while angina pectoris was more prevalent in Hispanic patients (87%). Diabetes and left ventricular hypertrophy were most common in black patients (33 and 29%, respectively), while hypercholesterolemia and prior revascularization (coronary artery bypass graft or angioplasty) were most common in Caucasian patients (64 and 41%, respectively). More than 60% of Hispanic and black patients were women--a unique characteristic for randomized CAD trials. Comparing race/ethnic cohorts, there were significant differences for all characteristics. More than 80% of patients in all race/ethnic groups were receiving antihypertensive therapy; however, only fewer than 25% had controlled blood pressure according to guidelines from the sixth report of the Joint National Committee on Prevention, Detection, Evaluation and Treatment of High Blood Pressure. CONCLUSIONS: This high-risk population of hypertensive patients with CAD has been undertreated and does not have well-controlled BP. Race/ethnic differences were observed for clinical characteristics and medication use. PMID- 15553311 TI - Dyslipidemia in patients with angiographically confirmed coronary artery disease- an opportunity for improvement. AB - BACKGROUND: There are few data about lipid profiles in unselected patients with angiographically confirmed coronary artery disease (CAD). HYPOTHESIS: The study was undertaken to investigate the demographics, clinical characteristics, angiographic findings, and baseline lipid status of 1,000 consecutive unselected patients with angiographically confirmed CAD. METHODS: Between April 2001 and July 2002, we obtained informed consent and prospectively collected clinical characteristics, fasting lipid profiles, and angiographic results from 1,000 sequential patients with CAD confirmed by angiography. RESULTS: In these patients with confirmed CAD, 78% had history of hyperlipidemia. Although 62% were receiving lipid-lowering therapy, only 46% had a low-density lipoprotein target of < 100 mg/dl, and only 20% had achieved all four National Cholesterol Education Program-recommended lipid targets. CONCLUSIONS: Better strategies to ensure optimal lipid levels are required. One such method using computerized workflow is being evaluated in this population. PMID- 15553312 TI - Intravascular ultrasound-guided cutting balloon angioplasty for renal artery stent restenosis. AB - Renal artery stenting is the procedure of choice for treating atherosclerotic renal artery stenosis; it improves the acute results of percutaneous balloon angioplasty and has a better long-term outcome, particularly a reduced risk of restenosis. However, the treatment of renal artery stent restenosis is not well established and it is challenging because of limited therapeutic options. A novel technique of treating renal artery stent restenosis using cutting balloon angioplasty guided by intravascular ultrasound is presented. PMID- 15553313 TI - Profiles in Cardiology. Ilya Fadeyevich Tsion, alias Elias Cyon, alias Elie de Cyon. PMID- 15553314 TI - [The importance of nurses in the professionalization of the nursing profession in Germany]. PMID- 15553315 TI - [Living with pain--elderly women as experts in the management on their chronic musculoskeletal pain]. AB - Chronic musculoskeletal pain in the elderly is very common. The aim of this qualitative study was to gain insight in elderly people's pain medication beliefs. Problem-focused interviews with eight women were conducted. The results show them to be experts in managing pain, which task turns out to be too complex to be described in terms of pain medication beliefs. Using the method of qualitative content analyses five main categories including subcategories were inductively generated and presented in a structured way. The category "to be carried by one's life and illness trajectory" evolved as a basic category in the pain management. It includes the subcategories "to gain experiences from one's life and illness", "to have knowledge of the causes of pain and of their treatment", "to learn how to manage pain in everyday life" as well as "to rely on spiritual well-being". Most important for the women is their every day reality "to live with pain and its physical, psychological, practical and social effects". This leads to "weigh, to combine and to evaluate treatments for pain relief" which includes the subcategories "to be disciplined in carrying out non pharmacological measures for pain relief", "to use pain medication sparingly but purposefully" and "to cooperate as a partner with health professionals". Some of the participants are challenged by "reaching their limits", which means "to reach the limits of endurable pain", "to experience the limits of failed treatment effects" as well as "to reach the limits of endurable treatment side effects". "To sustain one's quality of life in spite of pain" seems to be the aim of the elderly women's endeavour. The results of this study demand collaborative care in a partnership with elderly people with pain acknowledging their expertise. The results also ask for transdisciplinary efforts to support elderly persons with pain and for the development as well as the evaluation of self-management education programs. PMID- 15553316 TI - [Thought styles in nursing]. AB - Proceeding from the occupation with the question whether the tension that nursing staffs experience in correspondence with the conflict of theory and practical orientation can be made transparent the authors found no sufficient explanations in the current theories of professionalism. The theory of thought styles and thought collectives (Denkstile und Denkkollektive) of Ludwig Fleck offers a perspective that takes a look at the inside of nursing, at the nursing staffs. From the analysis of the works of Fleck three styles, or thought collectives have been determined, that can be met in nursing. The thought style that corresponds with the pre-professional phase of a nursing career is characterised by a mainly caritative understanding of nursing. The thought style that comes with professional nursing is based on standardised knowledge by experience without scientific foundation. The thought style that is related to a professionalism of nursing connects scientific insights, nursing knowledge by experience and hermeneutic understanding of cases. The consideration of the current situation in nursing shows two problems that can be explained from the perspective of the thought styles: disturbances in communication because of terms that are interpreted differently by different thought styles and resistance against the consequences of some processes of development relating to professional-political aspects of nursing (making nursing academical and more professional). The resulting strategies of action include a further development of a thought style of a professionalism of nursing to be able to deal with needing of care and the reflection of conflict situations on the background of thought collectives. PMID- 15553317 TI - [The nursing relationship--psychodynamic aspects]. AB - The relationship between nurses and their patients is influenced by subtle and often unconscious emotions which influence proper nursing. In this paper psychological concepts of development from psychoanalysis, baby-watching and attachment theory are enlisted to understand more deeply emotional situations in nursing relationships. After considering anthropological basics of nursing relations with the elderly and the mutual dependency of nursing and nursed persons are discussed. The limits of comparison between early childhood nursing and professional nursing relationships are mentioned at the end. PMID- 15553318 TI - ["And the suffering is also borne by the following generations"--an international literature study on the nursing care for Jewish patients]. AB - Jewish refugees from the Commonwealth of Independent States (CIS) are a constantly growing minority about whose care needs little is known in the scientific literature. Considerably more research has been done on the care needs of members of the Jewish faith outside the CIS, especially of those living in the USA, Israel and Canada. The ensuing international literature study was compiled as part of a pilot project promoted by the Robert Bosch Stiftung concerning the care needs of Jewish refugees from the CIS, with the aim of investigating available nursing experiences with the target group. The keywords "Juden", '"judisch", "Migranten", "jew", , "jewish" "migrants" and "nursing" were used for the analysis of altogether 67 articles from various data sources. The literature study generated the following internationally significant topics: the psychological and social situation and the resulting special features of the nursing care required by all of them viz: two generations of holocaust-survivors, their children, and Jewish migrants from the CIS; various religious orientations subdivided roughly into the categories orthodox, conservative and reform, in that each category represents different attitudes which affect nursing care; aspects of Jewish ethics relevant to nursing; the specifics of caring for gerontological Jewish patients; and socio-cultural aspects of palliative care. One of the most important results was that Jewish refugees from the CIS share in particular the experience of persecution with the holocaust-survivors and their offspring, which bears specific implications for nursing care. PMID- 15553319 TI - [Ethnographic study on the care requirements of Jewish migrants in Germany]. AB - Jewish refugees from the Commonwealth of Independent States (CIS) are a constantly growing minority in Germany about whose care needs little is known in the scientific literature. This article presents the results of a qualitative explorative study on the personal concepts of illness, health and nursing care of Jewish migrants against the background of their lives in the CIS and in Germany. Ethnographic data were gained and analysed on the basis of the Developmental Research-Sequence-Method (Spradley, 1979). The article aims at the presentation both of new data on the care needs of these people and, with these as their basis, of strategies for professional nursing care. PMID- 15553320 TI - [Nursing Informatics: state of the art and future of a subspecialty in nursing]. AB - Nursing Informatics is defined as a specialty that combines nursing science, computer science and informatics to capture, process, store and communicate data, information and knowledge from the nursing domain. Therefore, Nursing Informatics is understood as a subspecialty within nursing. In Germany, Switzerland and Austria, Nursing Informatics looks back to more than 10 years of experience in the field. A Nursing Informatics Framework is proposed to represent the scientific and practical work. It goes beyond a mere definition and forms an action space for showing present achievements and the need for future activities. An analysis of the state of the science reveals that applications in patient care, management and education and training are nearly equally covered by Nursing Informatics. However, a deficit in basic research is apparent mainly in formalising and representing nursing knowledge. There is also a lack of system descriptions following the phases of the software-engineering process. In the requirements specification phase for nursing information systems meaningful reporting applications for use in quality assurance, evidence based nursing and management will have to be identified. Systematic software-engineering including scientific evaluations is needed in particular of systems in emerging areas such as case management. However, the activities proposed require a thorough education and training in Nursing Informatics on all levels. PMID- 15553321 TI - [Concerning pharmaceutical care]. PMID- 15553322 TI - [Evaluation of methadone dispensing in Belgium--study synopsis]. PMID- 15553323 TI - [The follow-up of antiepileptic drugs]. AB - All antiepileptic medications have potential side-effects. Some are rather specific like diplopia for carbamazepin or lamotrigin, whereas others are not, like fatigue or unsteadiness. Most are dose-related and can therefore be alleviated by dose reduction (e.g. somnolence or tremor) but a few are idiosyncratic (e.g. rash) and require cessation of the causative agent. Some can be detected and followed-up on a clinical basis but others necessitate specific examinations. PMID- 15553324 TI - [Ototoxicity-related dysequilibrium]. AB - Many substances may be the source of dizziness or transient equilibrium disturbances due to dysfunction of the CNS or to an impairment of visual or proprioceptive informations. Other agents are responsible for drop of arterial pressure by changing position, including antihypertensive drugs, alpha-blocking agents used in urology, antipsychotics, cyclic antidepressants, vasodilators and nitrates, dopaminergic antiparkinson drugs, sedatives, etc. Only drug with true ototoxic properties will be discussed here, namely substances that are able to damage the inner ear (cochlear or vestibular damage) or the VIIIth cranial nerve, causing impairment of equilibrium and/or (most often) hearing. No relevant data report the actual incidence of ototoxic problems, but more than 130 products have been classified as potentially dangerous. Individual susceptibiity seems highly variable, but some predisposing factors have been identified: renal failure, age, combination of ototoxic drugs, familial sensitivity to ototoxic effects or previous neurosensorial deficit. We will first discuss the ototoxic medications that have certainly been extensively studied and among which we find several antibiotics (especially aminoglycosides and macrolides), the loop diuretics, and some antimalarial or chemotherapeutic agents. Environmental toxins and drug of abuse will then be discussed briefly because scientific data are much less significant. Early recognition of subjects who are at risk of developing ototoxicity, use of therapeutic monitoring and close observation of cochleo vestibular functions in high risk situations (often not easy in critical patients) are the best way to prevent severe complications that have occasionally disastrous consequences on the quality of further life. PMID- 15553325 TI - [Avian influenza ("bird flu") and its possible transmission to humans]. PMID- 15553326 TI - [Some new drugs: valdecoxib (Betra), voriconazole (Vfend), ezetimide (Ezetrol), teriparatide (Forsteo)]. PMID- 15553327 TI - [Prospective testing of nurses' identity]. PMID- 15553328 TI - [The QDF-HF (Quality of life, Depression and Cognitive Functioning in Heart Failure). The project and study protocol]. AB - The QDF (Quality of life, Depression Cognitive Function) is a project by nursing personnel, as companion study of the GISSI-HF (Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto miocardico- Heart Failure). The study aims at describing the evolution of the QDF variables in the patients randomised for the GISSI-HF; at evaluating the reliability of the assessments with standardised instruments (Kansas City Quality of life Questionnaire (KCQQ); Geriatric Depression Scale (GDS) and Mini Mental State Examination (MMSE) compared to the clinical judgement of the nurses; and at describing whether and how the assessment of QDF variables influences decision making and care for these patients (i.e. drug therapies, psychosocial interventions etc). The QDF data are collected at baseline and at 6 months and at 1, 2 and 3 years. The greatest majority of patients are expected to comply with the self administered questionnaires (KCQQ and QDF) while the MMSE will be proposed, via a didactical presentation, only to patients older than 70 years. A specific goal of the project focuses on the degree of independence between the results of the questionnaire and the judgement of nurses. Expected outcomes: epidemiology and distribution of QDF variables among patients of different age-groups and disease severity; description of the prevalence and incidence of severe problems. Up to 2000 patients are expected to be included in this prospective protocol which has already been activated in a network of more then 70 centres. PMID- 15553329 TI - [Doctors-nurses integration in paediatric onco-hematology: a cornerstone for quality of care. Report on a three years experience]. AB - The rapid evolution of care and the constantly changing needs and expectations of patients and their families require flexibility and re-organisation of the caring and treatment activities. These include the increased need for high-specialised personal competence; communication and interpersonal skills; and a close collaboration and integration between doctors and nurses--the main professional groups in the health care team. An optimal integration is not always and easily achievable and requires personal motivation, active efforts, investments and the development of the effective operative models for providing the integrated high specialised daily care. It is not easy in the busy daily routine to find time, motivation and share common goals. This is why a support for the groups willing to implement projects for improving integration between doctors and nurses was provided through an action-research laboratory. The aims of the multi-step action research laboratory were the following: to improve the integration of different professionals; to improve the exchange of knowledge and skills; to build the necessary bases for planning and running comprehensive, integrated clinical projects in the field of paediatric haematology-oncology. The ultimate goals of this effort was to improve the quality of the care and cure provided to children with cancer and their families. The project lasted three years, and consisted of yearly residential meetings alternating with experiences conducted in the working environments of the participants. The following steps were planned: Identification of the problem. Participants were asked to analyse the main problems of their units, that could be solved-improved with a better integration/collaboration between doctors and nurses and the main difficulties routinely encountered in working effectively together. Groups were asked to analyse the problem in their own practice, identify the possible solutions, main obstacles, strengths and weaknesses, plan the necessary of activities (short and medium term) for improving the problems. Sharing of projects. The projects were shared and common problems and solutions discussed. Final presentation of the projects with the aim of devising solutions and strategies for consolidation of the eventual changes obtained. Supervision, theoretical support and personalised feed-back from the scientific panel were guaranteed between the meetings and during the planning and implementation phases. Methods, instruments and techniques for improving collaboration and integration were discussed during the yearly meetings (i.e. how to work by objectives; how to plan and co-ordinate multidisciplinary meetings...) with lectures, workgroups, role-playing. Representatives from the 16 wards participated to the project: at least one nurse and one doctor from each centre, in the position to take decisions and implement changes. In spite of common problems related to the workload, difficulties in involving other colleagues, 13 projects were implemented. The need to discuss, plan together, organise interdisciplinary meetings improved the integration and communication between doctors and nurses. PMID- 15553330 TI - [A project to improve the collaboration between doctors and nurses in the care of terminal cancer paediatric patients]. PMID- 15553331 TI - [Integrated home care for paediatric cancer patients]. PMID- 15553332 TI - [The management of pain during invasive diagnostic procedures]. PMID- 15553333 TI - [Integrated communication between doctors and nurses at the first discharge]. PMID- 15553334 TI - [The set up of a phone on-call service at the oncohematology paediatric service]. PMID- 15553335 TI - [Criteria for the phone assessment of fever as a component of the phone on-call service]. PMID- 15553336 TI - [When a new nurse joins the team]. PMID- 15553337 TI - [The reorganization of the day-hospital]. PMID- 15553338 TI - [Continuity of care with written information and dedicated nurses: a literature review. Part 1]. AB - Continuity of care is one of the major challenges of contemporary care. The aim of this review is to evaluate the efficacy of written information and dedicated discharge nurses on continuity of care. A literature search was done on Medline (1966-March 2004) and Cinhal (1984-2004) with the following key words: "Community Health Nursing e Discharge Planning/Nursing Records/Continuity of Patient Care", "discharge liaison nurse", "liaison nurse", "transfer nurse" and "discharge coordinator (co-ordinator)" and "case manager nurse", a search by author was also performed. Editorials, letters and comments were excluded. Overall, 35 articles were identified. There are no evidences that written information improves the continuity of care and no agreement exists on the information to be registered: a diary of the patient hospital stay or detalied prescriptions of future interventions. A discharge professional seems to be effective on costs reduction, patients' satisfaction and according to some authors, also on the continuity of care. New and more extensive research is needed to shed a light on this pivotal aspect of care. PMID- 15553340 TI - Depression research: nursing's contributions. PMID- 15553339 TI - [A report on the international cooperative network for paediatric hemato oncology: the MASCOTA-MISPHO program]. PMID- 15553341 TI - Ethics. PMID- 15553342 TI - Improving the practice environment for evidence-based practice. PMID- 15553343 TI - Psychosocial theories of depression for individuals with intellectual and developmental disabilities: a historicist perspective. AB - The mental health care system has historically marginalized individuals with intellectual and developmental disabilities (I/DD). Until the mid-1980s, many clinicians doubted that individuals with I/DD were capable of depression (Sovner & Pary, 1993). Although it is now generally accepted that individuals with I/DD do have depression, they may not be treated or may be inappropriately treated (Matson et al., 2000). A historicist perspective takes into account the effect of marginalization on science and practice. Depression has both biological and psychosocial aspects. Key groups of theories regarding the psychosocial aspects of depression include psychodynamic/psychoanalytic, behavioral, cognitive, and ecological/interpersonal theories (Clark, Beck, & Alford, 1999; Joiner, Coyne, & Blalock, 1999). The application of psychosocial theories of depression to individuals with I/DD continues to reflect their marginalization and oppression. Behavioral theories of depression are limited in their conceptions for research, identification, and treatment of depression but continue to be used widely with individuals with I/DD. Cognitive theories of depression are widely used in research and treatment of depression in the general population, but have limited usage among individuals with I/DD. Interpersonal theories of depression are used in the general population and have many benefits, but are only now being investigated for use with individuals with I/DD. In this article, theories of depression as applied to individuals with I/DD are discussed from a historicist perspective. PMID- 15553344 TI - A longitudinal study of maternal postpartum depression symptoms. AB - The purpose of this study was to examine changes in maternal depression symptoms over time from the postpartum period to 2 years after delivery among a sample of women identified with elevated depression symptoms at 2 to 4 weeks postpartum, a longitudinal within-subjects design was used to examine changes in maternal depression symptoms. Two years after delivery, 62 women who had elevated depression scores at 2 to 4 weeks postpartum completed mailed questionnaires including a Demographic Information Sheet, the Beck Depression Inventory II, and the Parenting Stress Index. Among this sample of women, 30.6% scored in the depressed range 2 years after delivery. Although depression scores decreased over time, the significant change in mean scores occurred from 4 to 8 weeks to 10 to 14 weeks postpartum, and mean scores did not change significantly from 10 to 14 weeks, to 14 to 18 weeks, to 2 years after delivery. Depression history, lower overall social support, and higher parental distress were associated with higher depression scores among mothers at 2 years after delivery. Initial decrease in depression scores suggests that postpartum depression symptoms posed a time limited problem for many women with peak occurrence from 4 to 8 weeks after delivery. However, for many women depression symptoms persisted at 2 years after delivery. Previous depression, limited current support, and parental distress increased depression symptom severity. Additional longitudinal research is needed to determine factors that increase and ameliorate risk for chronic maternal depression. Clinical assessment of maternal depression beyond the early postpartum weeks is warranted for at-risk women. PMID- 15553345 TI - Prenatal depression: a randomized controlled trial in the emotional health of primiparous women. AB - The prevalence of postnatal depression (10%-15%) renders it a major public health problem not only for the depressed mother but also for the infant, who may suffer from behavioral disturbances and cognitive delays in later years. This study aimed at evaluating an educational intervention to alleviate postnatal depression and at generally measuring the prenatal and postnatal mood of primiparous women. A prospective, randomized controlled trial of an education intervention to reduce postnatal depression was conducted at three sites in Australia enrolling a total of 184 primiparous women. The intervention consisted of an information booklet on postnatal depression and an audiotape of one woman's journey through clinical postnatal depression. Mood was assessed once prenatally (12-28 weeks) and twice postnatally (8-12 weeks and 16-24 weeks) using the Scale for Assessment of Depression and Schizophrenia modified for pregnant and postnatal women (SADS-M). Demographic and social support data were also collected at enrollment. Comparisons between the control group and the intervention group revealed no differences; the educational intervention did not show any effect when women's mood was measured by the SADS-M. Overall, a general, significant, steady decrease of depressive tendencies was observed when the two postnatal assessments were compared to the prenatal measurements. Women were less depressed postnatally than prenatally. This overall improvement of mood was significant in most SADS-M items. The exceptions were discouragement, anxiety, anger, and irritability, which did not reach significance. Additional multivariate analyses revealed no relevant influence of social support or demographic variables on the changes in mood. The main results that the education intervention had no effect and women, overall, were more depressed prenatally than postnatally contributes further evidence to the view that the prenatal period is a separate entity from the postnatal period, with distinctive psychoneuro-endocrine pathways and, thus, suggesting different profiles of women's experience. This evidence indicates the necessity to screen, refer, and manage prenatal maternal mood as an entity in its own right, rather than as a window on the postnatal period. PMID- 15553346 TI - Reports on depressive symptoms in older adults with chronic conditions. AB - Depression is the most common mental disorder among older adults in the United States and one of the most disabling conditions worldwide. Chronic conditions and related functional limitations are associated with late-life depression, but assessment of depression is complicated by the absence of measures that capture the range of depressive emotions older adults may express. This descriptive, correlational study of 314 older adults with chronic conditions examined three measures to assess depressive symptoms: the Center for Epidemiological Studies Depression Scale (CES-D), the short form of the Center for Epidemiological Studies Depression Scale (CES-D-10), and an Emotional Symptom Checklist (ESC). The measures were correlated with each other and with a number of chronic conditions and functional impairments. Men and women scored similarly on all measures, though correlations between depressive symptoms and negative emotions were stronger for men. About 12% of the older adults exceeded the CES-D criteria for severe depressive symptoms, with the greatest percentage among those aged 75 to 84. The most frequently reported negative emotions were sadness (by women and elders through age 84) and loneliness (by men and elders age 85 and over). The findings suggest the need for multiple assessment strategies to identify older adults at risk for late-life depression. PMID- 15553347 TI - The impact of high and low stress on the health of Lebanese families. AB - During the Lebanese war (1975-1991) families were subjected to a substantial number of war and nonwar life events that were shown to have a negative impact on family adaptation. The study was undertaken to classify and predict family adaptation outcomes in high and low perceived stress groups for war and nonwar life events. The study took the form of an analysis of cross-sectional data of families in Beirut. The cluster sample consisted of 438 families chosen at random. Variables included perceived stress for war and nonwar events, and health and interactional indicators of the outcome family adaptation assessed in terms of physical and psychological health, depression, and interpersonal and marital relationships. Families were classified into high/low stress based on their score for war and non-war life events. Based on discriminant function analysis, a significant difference existed between the high- and low-stress groups (for war, nonwar, and a combination of the two) on all predictor variables. Interpersonal relationships, physical health, and depression were the best predictors of membership in the high/low war stress groups. In the perceived nonwar stress groups, depression, interpersonal relationships, marital relations, and physical and psychological health were the best predictors separating high/low stress groups. When the combination score of war and nonwar stress was used, interpersonal relationships, physical health, depression, and marital relations were the best predictors distinguishing among the groups. Findings indicated a high degree of consistency in the prediction and classification of the high/low stress groups and for all stress categories. Families in the low-stress groups had a more positive adaptation than those in the high-stress groups and could be classified on the basis of their stress score. Families in the high-stress groups had more complaints than their counterparts in the low-stress groups. This classification may help mental health professionals understand how families adapt in response to normative and non-normative life situations. PMID- 15553348 TI - Chronic stress conditions do explain posttraumatic brain injury depression. AB - Psychosocial and biologic mechanisms are implicated in depression after traumatic brain injury (TBI). Using McEwen's stress theory of allostasis as a guidepost, this study examined whether pre- and postinjury chronic stress conditions could explain post-TBI depressive symptoms. Seventy-five community-dwelling persons who sustained a mild-to-moderate TBI and were within 2 years of the injury participated in this cross-sectional study. The participants completed measures of chronic stress and depression, measured with the Neurobehavioral Functioning Inventory. Data were collected also on brain injury severity. Using multiple regression analysis, the frequency of childhood adversities and postinjury stress explained post-TBI depression. When time-since-injury was in the regression model, the frequency of preinjury stressors and postinjury stress significantly explained post-TBI depressive symptoms while the combined effect of childhood adversity with postinjury stress was not significant in explaining depressive symptoms. Pre- and postinjury chronic stress explained post-TBI depressive symptoms. These findings support stress-diathesis theory within the psychiatric literature and a linkage between chronic stress, an indicator of allostatic load, and post-TBI depression. These findings are important for nurse specialists working with persons who sustained brain injury, for chronic stress can be buffered by efficient and effective support systems. PMID- 15553349 TI - The relationship between maternal and child mental health in Mexican immigrant families. AB - Depression is a leading cause of disability in the United States, with Mexican immigrant women reporting depression rates higher than the national average. The purposes of this study were to describe mental health symptoms in a sample (n = 182) of Mexican immigrant mothers and their relationships to child mental health, family functioning, and acculturation. Over one third of the mothers reported depression and anxiety symptoms above standardized cutoffs while 31% of the children scored in the depressed range. Of those children with high depression scores, 51% also had a mother with high depression and anxiety scores. Boys' depression scores were related to maternal reports of family functioning and stress, while girls' depression scores were related to maternal reports of depression, anxiety, and stress. Maternal mental health symptoms were associated with family functioning but not with acculturation. These data indicate that poorer maternal mental health and family functioning is associated with greater stress in Mexican immigrant children. PMID- 15553350 TI - Predictors of depression in Thai women. AB - This cross-sectional correlational study examined the associations between depression and family history of depression, early loss of mother before age 11, number of stressful life events, women's role strains, interpersonal conflict, parental role strain, perceived stress, social support, and sense of belonging in 142 Thai women (77 depressed and 65 nondepressed). Results from chi-square tests and t tests revealed that compared to nondepressed women, depressed women were more likely to have a family history of depression and history of loss of mother before age 11; significantly higher stressful life events, parental role strain, interpersonal conflict, and perceived stress; and significantly lower interpersonal resources, perceived social support, and sense of belonging. Results from logistic regression, however, showed that only family history of depression, perceived stress, and sense of belonging predicted depression in Thai women. Results from path analysis showed that 66% of the variance of depression was explained by the proposed model. Perceived stress had the strongest direct effect on severity of depression. Family history of depression had both direct and indirect effects on severity of depression. Number of stressful life events, interpersonal conflict, parental role strain, perceived social support, and sense of belonging had indirect but not direct effects on severity of depression. Loss of mother before age 11 and interpersonal resources had neither direct nor indirect effects on severity of depression. Results from this study contributed significant information for understanding depression in Thai women and can help health care providers in Thailand to provide better care for depressed patients and prevent depression in Thai women. PMID- 15553351 TI - Depression coping self-efficacy as a predictor of relapse 1 and 2 years following psychiatric hospital-based treatment. AB - This longitudinal study involved 1 - and 2-year follow-up assessments of participants who received inpatient (n = 25) or partial hospital program (PHP) (n = 24) treatment for depression. The purposes of this study were to (a) compare 2 year follow-up depressive symptom and depression coping self-efficacy scores with posttreatment scores, (b) examine depression coping self-efficacy scores as a predictor of health care visit frequencies following treatment, and (c) examine differences in 2-year depressive symptom and depression coping self-efficacy scores between original treatment groups. Participant (n = 49) health records were reviewed for frequencies and types of health care visits at 1-year posttreatment. Twenty-six (52% response rate) participants completed the Depression Coping Self-Efficacy Scale, Center for Epidemiologic Studies Depression Scale, and Two-Year Follow-Up Questionnaire at 2-years posttreatment. One-year findings indicated posttreatment depression coping self-efficacy scores were associated with frequencies of psychiatric episodes of care. Relapse rates (> 25%) were consistent with previous reports. Despite maintenance of improvements in depressive symptom and depression coping self-efficacy scores, depressive symptom scores remained above cutoff levels at 2 years posttreatment. Two-year findings did not differ based on original treatment type. Findings support further research examining the DCSES as a predictor of relapse and a target for treatment. PMID- 15553352 TI - [The adrenocortical carcinoma, a tumour of wide clinical diversity]. AB - Over the course of a few years, an obese 52-year-old woman with a 23-year history of hypertension developed a number of abdominal complaints including gall stones. Her blood pressure became increasingly difficult to control and she developed diabetes mellitus and suffered palpitations and headaches. It became noticeable that she had a moon face. Laboratory tests revealed hypercortisolism. CT-scan showed a large inhomogeneous mass of nine centimetres in her left adrenal gland, which was subsequently removed surgically. The histopathological diagnosis was consistent with an adenoma. After a number of months the patient developed bone and liver metastases and the diagnosis was amended to carcinoma of the adrenal cortex. She then underwent radiotherapy and chemotherapy treatment. One year after surgery she developed a pancytopenia and died. Adrenocortical carcinomas are rare tumours with an incidence of about 1-2 cases per million of the population. Symptoms are heterogeneous since both functional (hormonal overproduction) and non-functional (mass effect) tumours exist. Surgical resection is the only curative therapy. It may be difficult to distinguish between benign and malignant cortical tumours. PMID- 15553353 TI - [The implantable cardioverter-defibrillator: sometimes necessary]. AB - The implantable cardioverter-defibrillator (ICD) is used in patients who are at risk for ventricular fibrillation after having suffered from a myocardial infarction. Initially, patient selection was limited to survivors of impending sudden death with coronary artery disease. Later, ICD implantation in high-risk coronary artery disease patients was found to lower cardiovascular mortality. More recently, patients with dilated cardiomyopathy and no coronary artery disease are also potential candidates for implantation of an ICD. In the Netherlands, it is expected that there will be 1-2 ICD-users per 10,000 inhabitants. This means that ambulance personnel, general practitioners and doctors in emergency wards will also be confronted with patients who have had one or more shock treatments. Such medical personnel should have knowledge about the function of the ICD, what to do if several shocks have been given and whom to contact in case problems persist. PMID- 15553354 TI - [New fluoroquinolones: levofloxacin, moxifloxacin and gatifloxacin]. AB - Since their introduction in the seventies, the fluoroquinolones have gained an important place in the treatment of Gram-negative infections. Modifications in their structure have led to a number of newer agents with improved activity against Gram-positive microorganisms. Among these new agents are gatifloxacin, levofloxacin and moxifloxacin, which have been registered in The Netherlands. These new fluoroquinolones are promoted by the manufacturers for use in respiratory tract infections, notably in community-acquired pneumonia. Studies have shown that these agents may be somewhat more effective than the current antibiotics in use in The Netherlands for community-acquired pneumonia. Given the fact that penicillin resistance among pneumococci hardly occurs in The Netherlands and the severe risk of the development of resistance in both Gram positive and Gram-negative microorganisms when the new fluoroquinolones are used, their area of indication would currently seem to be limited. PMID- 15553355 TI - [The practice guideline 'Photo(chemo)therapy and systemic therapy in severe chronic plaque-psoriasis']. AB - The guideline for the treatment of adults with moderate to severe chronic plaque type psoriasis vulgaris is based on published scientific evidence. The efficacy of the treatment, its safety, the side effects, the patients' discomfort, the patients' preference and the costs of treatment and follow-up are listed as criteria that play a role in the choice of treatment. Photo(chemo)therapy is the treatment of choice for severe forms of psoriasis. Phototherapy with ultraviolet B light (UVB), especially narrow-band UVB, is preferred above photochemotherapy with ultraviolet-A light and psoralenes (PUVA). Of the oral treatments, methotrexate and ciclosporin are preferred above retinoids and fumarates because they yield a higher percentage of patients with partial or nearly complete remission. Fumarates are not officially registered in The Netherlands and Belgium. PMID- 15553356 TI - [From gene to disease: amyotrophic lateral sclerosis]. AB - Amyotrophic lateral sclerosis is a progressive neurological disorder. It is characterised by selective motor-neuron degeneration in the cortex, brainstem, and spinal cord. Consequently, patients suffer from muscle weakness and usually die within 3-5 years after diagnosis from respiratory insufficiency. About 5-10% of the patients have a family history of ALS, the remaining are classified as sporadic ALS. There is only limited information about genetic susceptibility factors in sporadic ALS. Some patients with familial ALS have mutations in the gene encoding for copper/zinc superoxide dismutase, a protein involved in scavenging superoxide radicals. This results in a toxic gain of function. Mutations in the gene coding for alsin, ALS2, have been shown to be responsible for an autosomal recessive form of juvenile ALS. PMID- 15553357 TI - [Diagnostic image (212). A man with severe abdominal pain]. AB - A 36-year-old man presented with acute abdominal pain due to occlusion of the superior mesenteric artery associated with intestinal rotation caused by an upper abdominal postoperative adhesion. PMID- 15553358 TI - [The rise in fluoroquinolone-resistant Neisseria gonorrhoeae among people attending the Municipal Health Service's clinic for sexually transmitted diseases in Amsterdam, the Netherlands; cefotaxime now first-choice treatment for uncomplicated gonorrhoea]. AB - OBJECTIVE: To report the incidence of gonorrhoea and the development of resistance to Neisseria gonorrhoeae among attendees at the Municipal Health Service's STD-clinic in Amsterdam, The Netherlands, 2000-2003. DESIGN: Descriptive. METHOD: Urethral or cervical swabs for culture for N. gonorrhoea were taken from attendees at the STD-clinic. Depending on reported sexual techniques throat and rectal swabs were also taken. The disk diffusion technique in combination with a beta-lactamase test were used for sensitivity testing. RESULTS. The number of Neisseria gonorrhoeae (NG) isolates collected at the Amsterdam Municipal Health Service's STD-clinic decreased from 1047 in 2002 to 772 in 2003. The number of fluoroquinolone-resistant NG (FRNG) isolates rose from 3 in 2000 to 56 in 2003 (p < 0.001). FRNG isolates amongst men who have sex with men increased from 1 in 568 isolates (0.2%) in 2000 to 50 in 478 isolates (10.5%) in 2003 (p < 0.001). Amongst heterosexual men, FRNG rose from 2 per 275 (0.7%) in 2000 to 16 per 297 (5.4%) in 2002 and dropped to 6 per 190 (3.4%) in 2003 (p = 0.146). No FRNG isolates were found in women in 2003 (2000: 0/180 (0.0%); 2001: 2/160 (1.3%); 2002: 4/183 (2.2%). CONCLUSION: The recent incidence of FRNG among men who have sex with men to over 5% makes ciprofloxacin and other fluoroquinolones obsolete as the first-choice treatment option for uncomplicated gonorrhoea if no antibiogram is available. It is advised to use cefotaxim when an antibiogram is not available (yet). PMID- 15553359 TI - [Implantable cardioverter-defibrillator in the treatment of two patients with an increased risk of sudden cardiac death]. AB - A 23-year-old female with familial long-QT syndrome and a 48-year-old male with familial dilated cardiomyopathy were given an implantable cardioverter defibrillator (ICD) as prophylaxis. About half a year after the implantation, there was an appropriate and successful ICD-discharge in both patients in connection with ventricular tachycardia. Treatment with an ICD can be life-saving in patients with cardiac rhythm disorders. The most common indication is ventricular tachycardia or fibrillation due to ischaemic heart disease, but an ICD may also be indicated in patients with cardiomyopathy, congenital heart disease, hereditary arrhythmia or a planned heart transplantation. PMID- 15553360 TI - [Large-scale, acute, bacterial gastroenteritis caused by the enterotoxin of Staphylococcus aureus after a barbecue]. AB - EPIDEMIOLOGICAL OBSERVATION: On a warm day, during a barbecue that was attended by over 100 guests, a large number became rapidly ill with signs of acute gastroenteritis within a few hours after eating the prepared food. The characteristic symptoms were nausea, vomiting, syncope and in some cases in a later stage, diarrhoea. Sixty patients were transferred to hospitals. INVESTIGATION: Investigation revealed that the cause of this outbreak of gastroenteritis was an enterotoxin-A-producing strain of Staphylococcus aureus in a noodle dish. Both the food residues and the faeces from patients contained genotypically identical strains of S. aureus. They all had the gene for the gastroenteritis-inducing enterotoxin A from S. aureus. CONCLUSION: This case shows that the timely involvement of the different health authorities responsible for intervening in explosions of food poisoning is crucial for the clarification and treatment of such large-scale outbreaks. In the investigation of this outbreak, the municipal health authority co-operated with regional laboratories and the Food and Consumer Product Safety Authority/Inspectorate for Health Protection and Veterinary Public Health. PMID- 15553361 TI - [Concerns relating to care for transsexuals]. AB - In the last decade, transsexual patients have increasingly sought sex reassignment (cross-sex hormones and sex reassignment surgery), whilst avoiding adequate diagnostic procedures. They ask non-specialized physicians to prescribe hormones or to perform sex reassignment surgery. Sometimes hormones are ordered through the Internet or obtained from other illegitimate sources. If these patients later turn to a specialized team (for instance for sex reassignment surgery), the obligatory standard diagnostic procedure is problematic: the patient does not accept having to go through a long period of assessment and objective decisionmaking, while the attending health professional feels pressured by the patient's impatience. Non-expert health professionals should draw patients' attention to the necessity of a thorough diagnostic procedure, thus avoiding wrong decisions and future regrets. PMID- 15553362 TI - [Bile duct injury after cholecystectomy: risk of mortality substantially higher]. PMID- 15553363 TI - [Successful treatment of three elderly patients suffering from prolonged delirium using the cholinesterase inhibitor rivastigmine]. PMID- 15553364 TI - [Small-bowel obstruction as the expression of hereditary non-polyposis colorectal carcinoma]. PMID- 15553365 TI - [Diagnostic image (202). A newborn with subfebrile temperature and skin lesions]. PMID- 15553366 TI - [Digital radiography in young children. Considerations based on experiences in practice]. AB - In dentistry, digital radiology techniques, such as a charge-coupled device and a storage phosphor plate, are gaining popularity. It was the objective of this study to assess the importance of the advantages and disadvantages of digital radiology techniques for bitewing radiography in young children, when compared to conventional film. A group of dentists received a questionnaire regarding their experiences with digital radiology techniques or conventional films among young children. Using the Simple Multi-Attributive Rating Technique (SMART) a final weighted score was calculated for the charge-coupled device, the phosphor plate, and conventional film. The scores were 7.40, 7.38, and 6.98 respectively. The differences were not statistically significant (p > 0.47). It could be concluded that, on the basis of experiences in practice, there are no statistically significant preferences for the use of digital radioogy techniques for bitewing radiography in young children. PMID- 15553367 TI - [Acceptance and results of electronic implantology instruction]. AB - Aim of this study is the evaluation of a 3-weeks course on dental implants presented in a computer assisted learning program. Evaluation variables are study result, student satisfaction and the use of the program. Fourth grade dental students were divided in a group who received traditional education and a group who received the course by means of an interactive computerprogram. At the end of the experiment there is no difference in study result between the two groups. Students say that the computer assisted learning program is a useful tool in studying the course, but use the program more as a device to obtain information than as a tool to have interactive discussion. PMID- 15553368 TI - [Research methods in dentistry 4. Methods for evaluating the sealing ability of filling materials in the root canal]. AB - In the past many in vitro studies were presented on the efficiency of the apical closure of the root canal. Such studies are relatively easy to perform in contrast to clinical studies. In order to determine which type of apical seal will give the best result, many techniques are evaluated. These results from in vitro studies are difficult to apply to the clinical situation. The results of clinical studies on root canal treatments are not always in accordance with the findings of in vitro studies. It remains a challenge to design a research protocol with an improved predictive value for the clinical situation. PMID- 15553369 TI - [The role of health professionals in discouraging tobacco use]. AB - In Central and Eastern Europe the morbidity and mortality rates as a result of tobacco use are high. Based on the premise that the health professional can play an important role in dealing with this epidemic of tobacco use, a series of workshops was organized in 2000 and 2001 for dentists and physicians. This article contains a summary of the proceedings of the workshop 'The role of the health professional and health school curriculum in tobacco control', that have been earlier published in the European Journal of Dental Education. The role of the Dutch dentist in assisting their patients to quit smoking is discussed. PMID- 15553371 TI - [Differential diagnosis of multiple sclerosis and systemic vasculitis]. AB - Two groups, each included 12 patients, with confirmed diagnosis of multiple sclerosis (age 19-35 years) and with systemic vasculitis (age 21-47 years) were compared in the study. Differential clinical immunological and tomographical features of these two states are described. PMID- 15553370 TI - [An enamel disorder in two siblings]. AB - Two sisters exhibit a similar enamel disorder. It is a hereditary developmental disorder, diagnosed as amelogenesis imperfecta. In this article the various types of amelogenesis imperfecta are discussed and the clinical consequences, genetic aspects, and basic guidelines for treatment are addressed. PMID- 15553372 TI - [Higher cortical functions in children with spastic forms of cerebral palsy]. AB - Kinestatic praxis was studied in 75 children, aged 7-11 years, with spastic forms of cerebral palsy (CP) and in 30 of them other higher cortical functions were examined. The disturbances of kinestatic praxis were more expressed in hemiparetic forms of CP, being observed in all the cases, and frequently occurred in spastic diplegias (75-94.3%) as well. Dynamic praxis was disturbed in 90% cases, spatial praxis--in 53.3%, visual-acoustic gnosis--in 36.6%, visual-object gnosis--in 50%, visual-spatial--in 80%, visual memory--in 40%, visual imaging reasoning--in 30%, verbal logic reasoning--in 43.3%. PMID- 15553373 TI - [Cochleovestibular syndromes in vertebrobasilar insufficiency]. AB - The study aimed at (1) identification of the main vestibulometric characteristics of peripheral cochleovestibular syndrome caused by arterial hypertension, atherosclerosis and vegetative vascular dysfunction, (2) elucidation of the pathogenic peculiarities of its formation and (3) evaluation of therapeutic efficacy of betaserc in peripheral vertigo. Eighty-five patients with peripheral cochleovestibular syndrome (PCVS) were studied. On the base of the data of ultrasonic examination of major cranial arteries, arterial blood pressure, central hemodynamics and structural cerebral changes, it is shown that abnormalities of vertebrobasilar system play an important role in PCVS development. Stenosis of spinal and internal carotid arteries was found in a limited number of cases. A conclusion is made on involvment of hemodynamic mechanism of blood circulation insufficiency in the inner ear at microcirculating level in ischemic PCVS pathogenesis. A study of the structural cerebral changes revealed that PCVS were not accompanied by the ischemic foci in the brain. Pharmacotherapy of peripheral vertigo of vascular etiology is based on the right choice of the drugs optimizing blood circulation in combination with vestibulolytics. Therapeutic efficacy of 2-month betaserc course in peripheral vertigo has been proved. PMID- 15553374 TI - [Premorbid period of anorexia nervosa]. AB - Premorbid period of anorexia nervosa was studied in 96 female patients. In 64 cases (group 1) anorexia nervosa was as a separate disease and in 32 cases (group 2)--as a syndrome of endogenous mental disorder. In group 1 premorbid period was characterized by the features of neuropathy and residual organic disorders in combination with accumulation of subjects with personality disorders in the patient's family that promoted development of prolonged neurotic disorders. In group 2 there was hereditary psychosis load, high frequency of obstetrical and other perinatal complications, overvalued ideas and predisposition to autistic games and fantasying. In both groups, psychopathological peculiarities included typical affective, obsessive-phobic, hypochondriac, cenesthopathic and behavioral disorders, of a stable character often provoked by psychotraumatic events. PMID- 15553375 TI - [Cost-effect estimation of dopamine agonist (mirapex) application in patients with Parkinson's disease]. AB - Parkinson's disease is a neurodegenerative disorder with a growing social significance. Currently, dopamine agonists are considered as first-line medication used before levodopa treatment. High cost of dopamine agonists makes it necessary to estimate their cost efficacy by means of pharmacoeconomical studies. We studied efficacy of mirapex evaluated by UPDRS and PDQ-39 scores before and after 1-year treatment in 44 patients with Parkinson's disease: 18 patients non-pretreated with levodopa and 26 patients pretreated with levodopa. Treatment cost was estimated separately for two patients groups with subsequent cost-effectiveness analysis. Early use of mirapex as a dopamine agonist for patients non-treated with levodopa seems to be beneficial from clinical well as from pharmacieconomical points of view. PMID- 15553376 TI - [Functional myoelectrostimulation as a method for motor functions rehabilitation]. AB - The main features, namely diagnostic, prognostic, therapeutic, of functional myoelectrostimulation (FMES) method are described. FMES is primarily indicated in muscle function deficit, which can be organic (neuromuscle disorders) and functional (muscle relaxation). Five most essential FMES operations include: (1) choice of correctable movements and stimulated muscles; determination of (2) amplitude and temporal stimulation programs; (3) stimulation regimes in pathological gait; (4) stimulation parameters; (5) localization of the skin electrodes on the body. A positive influence of FMES on the process of movement rehabilitation was demonstrated in 2 cases of severe central nervous system pathologies--the consequences of the spinal cord lesion in sacral-lumbar area and hemiparesis of cerebral genesis. PMID- 15553377 TI - [Pronoran in the treatment of mental organic syndrome in the elderly]. AB - Efficacy and tolerability of pronoran (piribedil) was investigated in an open clinical trial including 21 patients, aged 61-85 years, with initial cognitive disturbances of cerebral vascular genesis. Along with a clinical and somatic examination, the Clinical Global Impression (CGI) scale, the Mini Mental State Examination (MMSE) test, the Sandoz Clinical Assessment Geriatric (SCAG) scale and a scale for side-effects were used. Pronoran was prescribed in a dosage 50 100 mg/day during 3 months. Patients were examined at base line and on the treatment day 14, 42 and 90. A positive clinical effect on mild cognitive impairment was achieved after 1.5 months and maintained to the end of 3 month therapy. Pronoran improves ability to sustain attention, learning and memory, organization of thinking and executive mental function. It also has a mild antidepressive and antianxiety action, being safe for elderly patients. PMID- 15553378 TI - [Polymorphism of electroencephalographic pattern in benign epileptiform discharges in childhood]. AB - Incidence and significance of benign epileptiform discharges of childhood (BECD), or rolandic spikes, have been studied in 2723 children, aged 2-15 years, with (841 patients) and without (1882) epilepsy. All the patients underwent standard electroencephalographic (EEG) study with video-EEG monitoring made in cases of epileptiform abnormalities. In the non-epileptic group, BECD frequency was 1.33%. There was a significant predominance of boys in both groups. The age of maximal BECD expression was 4-5 years in the non-epileptic group and 9-10 years--in children with epilepsy. The majority of children without seizures demonstrated different neurological and neurocognitive abnormalities, such as chronic headaches (25%), attention deficit hyperactivity disorder (25%) and speech delay (21%). Epileptiform abnormalities were observed mostly in the right hemisphere in children with epilepsy and in the left hemisphere in non-epileptic patients. There was a morphological similarity of electroencephalographic patterns in patients with rolandic epilepsy, benign occipital epilepsy, pseudolennox syndrome, Landau-Kleffner syndrome and electrical status epilepticus during slow sleep. The authors conclude that BECD are nonspecific feature of rolandic epilepsy and can occur in the broad spectrum of disturbances forming "hereditary impairment of brain maturation" group. Hereditary mechanisms involved in realization of various electroclinical features of focal brain dysfunction are suggested. PMID- 15553380 TI - [Hereditary motosensory neuropathies in Saratov region]. PMID- 15553379 TI - [D3 dopamine receptor gene Ser9Gly polymorphism in Russian patients with schizophrenia]. AB - Polymorphic marker Ser9Gly of dopamine receptor D3 gene is considered perspective for associative studies of schizophrenia. Allele and genotype frequency of this polymorphism were studied in different ethnic groups of schizophrenic patients as well as the attempts have been made to reveal an association with clinical presentations of the disease. However, the results are inconsistent. The present study aimed at investigating Ser9Gly DRD3 gene polymorphism in Russian sample of schizophrenic patients. One hundred and fifty patients with ICD-10 diagnosis of schizophrenia (broad definition), 69 male and 81 female, aged 34.8+/-13.87 years, age at disease onset 24.3+/-9 years, have been examined. Control group consisted of 150 healthy subjects without family history of schizophrenia, 60 male and 90 female, aged 32.7+/-13.5 years. No between-group differences have been found for Ser9Gly DRD3 allele and genotype frequencies. However, a frequency of homozygous genotype Gly/Gly was significantly higher in female patients, comparing to female controls (p=0.038 Yate's corrected, OR 9. CI 0.95% 1.0-79.5). A role of sex dependent association between Ser9Gly DRD3 polymorphism and schizophrenia is discussed. PMID- 15553381 TI - [Coaxil efficacy in depression in patients with chronic cerebral ischemia]. PMID- 15553382 TI - [Psychophysiological peculiarities of neurotic disorders]. PMID- 15553383 TI - [Nervous and mental diseases in boys and mutations in MECP2 gene]. PMID- 15553384 TI - [Neuroradiologic model of different types of neuronal migration dysfunction]. PMID- 15553385 TI - Love's burdens. PMID- 15553386 TI - "Allow natural death"--not so fast. PMID- 15553387 TI - A viable alternative to traditional living wills. PMID- 15553388 TI - Canadian pharmaceuticals. PMID- 15553389 TI - Canadian pharmaceuticals. PMID- 15553390 TI - Hijacking prescriptions. PMID- 15553391 TI - Influenza pandemic preparedness: legal and ethical dimensions. PMID- 15553392 TI - Deciding for the patient. PMID- 15553393 TI - Piercing the veil of corporate secrecy about clinical trials. PMID- 15553394 TI - Pharma goes to the laundry: Public relations and the business of medical education. PMID- 15553395 TI - FDA and the life-sciences industry: business as usual? PMID- 15553396 TI - The bioethics of business: rethinking the relationship between bioethics consultants and corporate clients. PMID- 15553397 TI - Impatient proponents: What's wrong with the California stem cell and cures act? PMID- 15553398 TI - Stem cell research should be more than a promise. PMID- 15553399 TI - Being a burden: reflections on refusing medical care. AB - Close relationships can give us reasons to care for others, even when the care is a burden. They can also give us reasons to accept care--even when accepting care means becoming a burden. PMID- 15553401 TI - [Secretary memory 2003]. PMID- 15553400 TI - Britain permits controversial genetic test. PMID- 15553402 TI - [The beginning of the first principles: the anthropic principle]. AB - The nowadays classical Anthropic Principle is put both in the historical perspective of the traditional problem of "the place of man in the Universe', and in the confluence of several scientific "border" issues, some of which, due to their problematical nature, are also subject of philosophical analysis. On the one hand, the scientific uses of the Principle, related to the initial and constitutional conditions of "our Universe", are enumerated, as they are supposedly necessary for the appearance and consequent development of Life--up to Man--. On the other, an organized collection of the principles of today's Physics is synthetically exhibited. The object of this work is to determine the intrinsic scientific nature of the Anthropic Principle, and the role it plays in the global frame of the principles of Physics (Astrophysics, Astrobiology and Cosmology). PMID- 15553403 TI - [Poverty, social exclusion, social capital and health]. AB - Social capital is the social structure which facilitates the actions of individuals, stimulates production and allows for success. Poverty maintains basic needs unmet (food, health, autonomy) over time and unvoluntarily. Social exclusion does not allow individuals to participate in society. The following dimensions are assessed: financial poverty, social inclusion, employment, health and education. Social participation, work integration, empowerment, self-esteem, and personal achievement should be promoted. In Europe 15% of people is exposed to poverty; in Spain corresponding figures are 13.4%, while for the elderly reached 21%. Extreme poverty affects 6.2% population and severe poverty 14.2%. Women and those living in Andalusia, Canary Islands and Extremadura are particularly affected, health inequality are for elderly, immigration, gender, social class, and should be reduced 10% for 2010. The Gini indez measures the income distribution; in the European Union (EU) it is 0.29 while in Spain is 0.33. Poverty and health are inversely correlated, health care expenditure in Spain is 7.5% og GDP. Life expectancy in U.E. is 75.5 years for men and 81.6 years for women, while in Spain it is 78 and 83.1 respectively. Infant mortality in EU is 4.5/1000, 4.1 per thousand in Spain. Lastly, the number of children per women in EU is 1.47 and in Spain 1.3. PMID- 15553404 TI - [The tortuose path of human evolution]. AB - The human genome sequencing and also the chimpanzee's, among others alive beings in every kingdom, and the similarities between genomes, due to an unique life origin, has developed two movements in the Biological Sciences to the XXI Century. In the first place to find a genetic explanation to understand the genomic affinity, very narrow in many cases, in front of the reality, with abyssal differences in behavior, which undervalue the significance of the genomic resemblance between the alive beings. The second movement is directed to ethological studies of the animal behavior and conduct, as a real fact. Special interest has the comparative studies since the two points of view, the genome and proteome in one side and ethology in front, both related to the Homo sapiens sapiens and his relatives, the great anthropomorphic apes, and consequently beside the names of Craig Venter, Collins and collaborators, artificers of the human genome sequencing, appear, unde revaluation, those of Leakey, Goodall, Fossey and Galdikas, specialists in anthropology and ethology of chimpanzees, gorillas and orangutans. These ideas are complementary in order to a better understanding of the origin, human evolution and behavior, shedding light on human cognitive abilities. PMID- 15553405 TI - [Sexual harassment]. AB - A study of the sexual pursuit is made insisting in the characteristics of the same one, in the consequences about the health and very especially on the etiology in which the prevention is based. PMID- 15553406 TI - [Rare diseases: a new chapter in medicine]. AB - Rare Diseases (RD) form a new chapter of Medicine which consists of more than 5.000 different diseases. RD have low prevalence (less than 5/10.000), chronic and severe evolution and usually produce serious alteration of quality of life and relationships with physical and social environment. The scarce number of patients with each RD produces very limited interest in research, specially in theraphy. "Orphan drugs" are useful drugs for treating some RD, and in spite of this they are not made because the low number of patients. The particular problem of one RD can be considered not important but as a whole it has great importance, having in mind its consequences over the patient, family and society. It is thought that there are about 20.000 patients in Spain and between 20 and 30 millions in the EU. Care of RD must be multidisciplinary (general practitioners, pediatricians, experts in RD, sociologists and social workers) with a permanent linkage between primary and specialised medicine. An appropriate training of professionals that work with these diseases is needed, as well as to increase and improve the research about RD and to give support to the patients and their families. Global Associations of patients with RD (FEDER in Spain, EURORDIS in EU and NORD in USA) as well as particular associations of some RD (more than 60 in Spain) have developed a very important task to improve health care and social attention about RD and must receive government and social support. PMID- 15553407 TI - [Effects of hormones on aging of the CNS, the immune system and the skin in rats]. AB - In a previous communication to the RANM two years ago, some data were presented supporting the role of GH, melatonin and estrogens in the prevention of aging of bone, liver metabolism and vascular activity. Additional data were presented about the beneficial effect of these hormones on the Central nervous System (CNS). In the present communication results obtained since that time on the CNS, the immunitary system and the skin will be presented. A total of 140 male and female rats have been investigated. Animals have been decapitated every 6 months starting with 2 and finishinf with 24 months. Another group of 64 rats from both sexes has been submitted to different treatments over 10 weeks, between 22 and 24 months of age. Males have been treated with GH and melatonin. Females were divided in two groups: intact and castrated at 12 months of age. The first group was treated with GH and melatonin and the second with these two and additionally with estradiol and with Phytosoya. Total number of neurones in the hilus of the dentate gyrus has been counted in Nissl stained cerebral cuts. Neurogenesis was estimated by staining with BrdU. Immunitary function was estimated measuring NK activity, chemiotaxis and lymphoproliferative activity in response to Concanavaline A in cultures of leucocytes obtained from axillary nodes and spleen of the animals as well as liberation of IL2 to the medium. An age dependent reduction in the number of neurones has been registrated in male and female rats, without influence of the ovariectomy. A dramatic reduction in neurogenesis has been also detected in both sexes. GH stimulates total number of neurones but does not modify neurogenesis, so a reduction in apoptosis has been considered. This seems to be supported by the reduction in nucleosomes and the increase in Bcl2 observed in cerebral homogenates. Immunitary functions show an age dependent reduction that gets worse with ovariectomy. GH treatment enhances all affected functions as well as estrogens and in some cases melatonin. The skin is also deteriorated by age and that is translated in a reduction of epidermal thickness, an increase in dermical fat and a lowering in the numer of ibroblasts. All these alterations are recovered with GH and melatonin. Keratinocytes from old animals in culture show an increase in nucleosomes nd a reduction in Bcl2 that is restaured with GH and melatonin. PMID- 15553408 TI - For injury hiding, athletic comparisons don't wash. PMID- 15553409 TI - Clarifying the rating of class B extinguishers. PMID- 15553410 TI - Sick leave item was misleading. PMID- 15553411 TI - Computer applications. The eyes have it. PMID- 15553412 TI - Migraine management. PMID- 15553413 TI - Raising the bar on hot work fabrics. PMID- 15553414 TI - Turning up the heat in emergency fixtures. PMID- 15553415 TI - Proper emergency equipment selection...there's more to it than meets the eye! AB - When we design and specify emergency equipment installations, we sincerely hope they are never needed. However, the reality is they probably will be used, no matter how safety-conscious a company may be. So, we logically seek out the best products, place them in the most appropriate places, and diligently train employees to ensure the maximum safety net is provided. As we've shown, there are differences in products that make some better for specific applications than others. And, as always, budget plays an important part in the selection. In the end, specifying becomes a balancing act--balancing features and capabilities against cost. The best way to deal with a balancing act is to know all of the facts! PMID- 15553416 TI - Maintaining emergency systems. PMID- 15553417 TI - Hazard assessment, eye protection, and you. PMID- 15553418 TI - Safety eyewear review. PMID- 15553419 TI - An IAQ overview. Increasing ventilation is a fundamental method to reduce concentrations of several substances of concern. PMID- 15553420 TI - Gas detection for VOC measurement. PMID- 15553421 TI - Why BBS really works. AB - Behavior-based safety does work. That has been proven over and over again, thousands of times. Why it works may not have been what they told you. Yes, the soon, certain, positive consequences will have an effect (a positive effect) on some deliberate behaviors, especially behaviors relating to personal protective equipment. But behavior-based safety or an observation and feedback process does much more than just provide soon, certain, positive consequences for safe behavior. Behavior-based safety also improves habits and increases awareness (eyes and mind on task). It takes hard work and skilled observers. The process must also be well managed and promoted. As mentioned before, it is hardly "free." But the rewards for all of this training and effort are huge. Injury reductions of 60-90 percent are nothing to sneeze at. If you had any doubt or were skeptical about the theory, I hope this has helped somewhat in terms of clearing up what really works and what doesn't in a behavior-based safety process. However, one thing that is not in doubt: Observation and feedback processes do work. That is why all of the safest companies in the world use them-not because they're expensive, time consuming, and must be actively managed, and not because these companies have a lot of spare time and extra cash. No, it is because observation and feedback processes reduce injuries; not by a little, but by 60 to 90 percent. PMID- 15553422 TI - Defibrillators & CPR. Safe & secure. PMID- 15553423 TI - Follow through experiences--as midwifery curriculum. PMID- 15553424 TI - Nurturing the future of midwifery through mentoring. AB - Mentoring has become a popular approach for providing support in developing knowledge and practice across a variety of disciplines. Midwives often provide mentoring for midwifery mentees and newly graduated midwives without formal recognition or predetermined outcomes. Little literature exists within midwifery practice outlining how its use may be of value. This paper presents literature around the topic of mentoring, discussing possible benefits and suggesting ways in which mentoring may be utilised within midwifery to promote personal and professional development. Finally, it presents some of the limited examples of mentoring emerging from the midwifery literature. PMID- 15553425 TI - How midwives learn about breastfeeding. AB - Little is known about how midwives learn about breastfeeding. This study asked midwives to identify breastfeeding information resources used and perceived value for their learning. A mail questionnaire was sent to midwives (n = 3500) through the Australian College of Midwives Inc. (ACMI). A response rate of 31.6% (n = 1105) was obtained. On-the-job experience was the most common source accessed and continuing education the most valuable. Very few respondents (3.1% n = 34) acknowledged either their hospital or university midwifery education program as a valuable breastfeeding information source. There is scope for continuing education programs to address evidence-based lactation and infant feeding information. Midwifery curricula need to teach in-depth knowledge of human lactation and develop clinicians' skill base to assist breastfeeding women. The development of national standards for course accreditation on lactation and infant feeding by ACMI, Baby Friendly Hospital Initiative (BFHI) and Australian Breastfeeding Association (ABA) would be a useful quality measure. PMID- 15553426 TI - Late motherhood: the experience of parturition for first time mothers aged over 35 years. AB - Australia, like other developed nations, is exhibiting a trend towards later childbearing. At present, the birth rate of women aged 35 years and older is growing faster than for any other age group. Approximately 35% of mothers aged >35 years are first-time mothers, and it is commonly held that these women have particular issues of adjustment to motherhood. This longitudinal qualitative study entitled 'Transition to motherhood for first-time mothers aged 35 years and above' explores their experiences. In-depth interviews were conducted with 20 primiparae aged >35, over three junctures: 35-38 weeks gestation; 7-10 days postpartum; and 6-8 months postpartum. Preliminary findings from interviews conducted immediately postpartum are presented in this paper. Participants identified adjustment to motherhood as a particularly challenging event. PMID- 15553427 TI - Blood loss at delivery: how accurate is your estimation? AB - This paper will examine the literature on blood loss at delivery and report the findings of a small pilot study where midwives and doctors were asked to estimate blood loss at simulated stations. The results were analysed using simple frequency distribution. This simple study demonstrated that midwives and other health professionals underestimate blood loss at delivery by 30-50%. The implication for midwives is that there is a need to double their estimated blood loss at delivery when it is over 500 ml. PMID- 15553428 TI - [Laparoscopic pancreatic resection: preliminary experience]. AB - The aim of the study was to evaluate the results of laparoscopic pancreatectomy for pancreatic tumours. Four women and three men underwent laparoscopic pancreatectomy and were recruited into the study retrospectively over the period from June 2002 to February 2004. Pancreaticoduodenectomy (n = 4), intermediate pancreatectomy (n = 1) and distal pancreatic resection with splenectomy (n = 2) were successfully performed. Operative mortality was nil. The postoperative morbidity included two low-output pancreatic leaks. Mean operating time, blood loss and hospital stay were 342 minutes, 289 mL and 14 days, respectively. The pathological diagnosis was ductal adenocarcinoma in one, neuroendocrine tumour in five and metastatic melanoma in one. All patients are still well after a median follow-up of 7 months (range: 1-20 months). The patients appear to benefit from laparoscopic pancreatectomy for pancreatic tumours. The minimally invasive approach ensures adequate treatment but requires the expertise of highly skilled laparoscopic surgeons. PMID- 15553429 TI - [Treatment of acute biliary pancreatitis: our experience]. AB - The results of endoscopic and surgical treatment in 21 patients affected by acute biliary pancreatitis are reported. All patients were managed according to the same protocol. Once the diagnosis was formulated (pain, serum amylase, ultrasonography, cholestasis and/or cholangitis) and the severity evaluated (Ranson's criteria, glucose and urea levels), the 21 patients underwent endoscopic retrograde cholangiopanctratography--within 24 hours in 8 patients (predicted severe disease) or within 72 hours in 13 patients (predicted mild disease). Endoscopic sphincterotomy was performed in 18 patients with stones in the common bile duct and was successful in 94.5% (17 patients: 3 patients underwent more than one session to confirm or achieve clearance of the duct). The morbidity rate associated with endoscopic sphincterotomy was 16% (3 patients: 2 haemorrhages, 1 perforation). There were no deaths. Cholecystectomy was performed in 16 patients within 2-9 weeks of the initial attack of acute biliary pancreatitis. There were 3 minor complications (respiratory, laparotomy, urinary infection), no major intraoperative or postoperative morbidity, and no mortality. Acute biliary pancreatitis is still associated with significant morbidity and mortality, but can be managed safely with combined medical, endoscopic and surgical treatment. PMID- 15553430 TI - [Zuckerkandl's tuberculum: could it be useful in thyroid surgery?]. AB - A thorough knowledge of thyroid anatomy could reduce the incidence of lesions to the inferior laryngeal nerve. In view of its relationship with the recurrent laryngeal nerve and the parathyroid gland, Zuckerkandl's tuberculum should be considered an anatomical landmark for the recurrent laryngeal nerve in thyroid surgery. The aim of the study was to verify whether the identification of Zuckerkandl's tuberculum could be useful to reduce the incidence of recurrent laryngeal nerve lesions. Four hundred and thirty-two patients underwent thyroid surgery over the period from January 2001 to December 2003 for benign (377 patients) or malignant disease (55 patients). Three-hundred and forty-eight (81%) underwent total thyroidectomy. Zuckerkandl's tuberculum was found in 74.5% of patients, with a high prevalence in the right lobe: in 5% of patients it was grade I, in 50% grade II and in 45% grade III. Its presence was associated with the recurrent laryngeal nerve in almost all cases. Eight of the patients undergoing total thyroidectomy suffered recurrent nerve paralysis, only 4 of which proved definitive. Identification of Zuckerkandl's tuberculum allows safer isolation of the recurrent laryngeal nerve and superior parathyroid gland dissection. PMID- 15553431 TI - [Hypoparathyroid risk after total thyroidectomy]. AB - From January 1970 to December 1999, 881 patients with thyroid pathology underwent surgery consisting in 551 subtotal thyroidectomies and 330 total thyroidectomies. Permanent hypocalcaemia was present in 32 patients (3.6%). The importance of accurate isolation and ultraligature of the branches of the inferior thyroid artery in the prevention of parathyroid damage is stressed. PMID- 15553432 TI - Localization of the sentinel lymph node in breast cancer: prospective comparison of vital staining and radioactive tracing methods. AB - The aim of the study was to evaluate possible differences in accuracy between the radioactive tracing and vital staining method in the search for sentinel nodes in patients with breast cancer. From January 1999 to December 2000, 102 patients with T1 N0 breast carcinoma were recruited into the study for localization of sentinel nodes with vital blue dye staining and radioactive tracing and were then submitted to lumpectomy and axillary dissection. For the two methods, we estimated the percentage of sentinel nodes localized, the false-negative rate, the predictive negative and positive value and the accuracy. The vital blue dye staining method permitted localization of the sentinel node in 73% of patients with a false-negative rate of 8%, a predictive negative value of 92% and 92% accuracy. The radioactive tracing method permitted localization of the sentinel node in 97% cases with a false-negative rate of 0%, a predictive negative value of 100% and 100% accuracy (P<0.0005). The method that offers the better results is radioactive tracing. Currently, many authors use both techniques, since, in common practice, staining helps to identify the sentinel node with the probe. PMID- 15553433 TI - [Complications of open prosthetic surgery for large incisional hernias]. AB - The authors examine the complications of open prosthetic surgery for incisional hernias and analyse them from different points of view. The case series includes, over a period of 16 years, 492 hernioplasties for incisional hernias and 160 (32.5%) for large incisional hernias. The authors stress that the best results have been achieved by the retromuscular placement of prostheses. They conclude that while, on the one hand, the successful use of prosthetic material has marked the end of the inoperability of this pathology, such procedures are closely associated with the development of complications, which were unknown in the pre prosthetic era and are sometimes very serious and linked to the site of the prosthesis and to the kind of material used. PMID- 15553434 TI - [Small bowel metastases from lung cancer]. AB - Lung cancer is usually diagnosed at an advanced stage and metastases are present in 50% of patients. Small bowel metastases from lung cancer are rare, being more frequent in patients with melanoma, uterine, ovarian, kidney or gastrointestinal cancer, or osteosarcoma. From November 1998 to August 2003, 740 cases of lung cancer (641 non-small-cell lung cancer and 99 neuroendocrine tumours) were diagnosed. We also observed 64 patients with malignant pleural mesothelioma and performed 23 pleuropneumonectomies. Over the same period we admitted 4 patients (one recurrent) with small bowel metastases, three from lung cancer and one from malignant mesothelioma. The clinical symptoms were bowel occlusion and intestinal bleeding. Radiological techniques such as small bowel enema and CT enteroclysis were used with positive results. In one patient with intestinal bleeding capsular endoscopy revealed a bleeding metastasis. All patients were operated on. Neither mortality nor morbidity were observed. All patients were discharged after a median stay of 10 days. One patient is still alive and disease-free 39 months after the first intestinal surgery for metastases. Intestinal metastases from lung cancer are rare and the diagnosis is often late. In some cases the clinical manifestations of the metastases are observed before those of the primitive tumour. However, in the presence of small bowel occlusion and intestinal bleeding of uncertain origin, clinical history-taking is very important and diagnostic procedures must be performed to exclude a secondary pathology. PMID- 15553435 TI - [Laparoscopic transhiatal oesophagectomy: our experience]. AB - The authors present their initial experience with minimally invasive surgery of the oesophagus. Two emblematic cases of benign and malignant oesophageal pathology, treated by laparoscopic transhiatal oesophagectomy, are reported. The surgical procedure is described in detail and compared with other techniques reported in the literature. Its advantages in terms of postoperative pain and morbidity are stressed. The role of the procedure in neoplastic diseases of the oesophagus is still debated, above all with regard to the accuracy of the mediastinal node dissection. PMID- 15553436 TI - [Palliative endoscopic treatment of pancreatic cancer]. AB - Pancreatic cancer has a very poor chance of being radically resected (20-25%) at the time of diagnosis. Palliation has thus proved to be the mode of treatment adopted in the majority of cases. When a radical surgical resection is not feasible, the endoscopic approach is one of the most common options among the therapeutic methods available. Endoscopic palliation for the treatment of jaundice is regarded as the best choice. It is associated with very low morbidity and no mortality. Hospitalization is short and the treatment is also inexpensive compared to other procedures. The authors report on their experience with endoscopic palliation for the treatment of inoperable pancreatic cancer. PMID- 15553437 TI - [Emergency surgery for colorectal cancer: our experience]. AB - Colorectal cancers have an extremely negative prognosis in the elderly, with a high percentage of clinical presentations requiring emergency surgery and high perioperative mortality rates. The clinical manifestations of this type of cancer set in a cutely in 20% of cases due to the sudden onset of one of the basic complications. The authors report on their experience with 79 patients undergoing emergency surgery for colorectal cancer. The choice of operation was made on the basis of the patient's general condition, as estimated by his or her ASA score, and the presence of associated diseases. PMID- 15553438 TI - [Endoscopic treatment of colorectal polyps in a digestive endoscopy outpatient department]. AB - The aim of this report was to evaluate the effectiveness of the endoscopic treatment of colonic polyps to allow secondary prophylaxis in order to prevent the onset of cancer arising from adenomas. From October 2002 to January 2004 we performed 487 colonoscopies on a patient group with the following indications: screening prior to kidney transplant; screening for colorectal cancer (patients positive at faecal occult blood testing); follow-up of patients who had undergone colonic resections for colorectal cancer; patients with other diseases. Colorectal polyps were diagnosed in 15 males and 15 females, with a mean age of 63 years. All the neoplasms were resected during colonoscopy and specimens sent for histological study. The histological examinations yielded the following results: 4 hyperplastic polyps; 9 tubular adenomas (6 with mild, 2 with mild-to moderate, and 1 with severe dysplasia); 8 tubulo-villous adenomas (3 with mild, 1 with mild-to-moderate, and 4 with moderate dysplasia); 4 villous adenomas (3 with mild and 1 with severe dysplasia); 1 adenocarcinoma; 1 inflammatory polyp; in 3 cases we were unable to retrieve the polyps after polypectomy. Colonoscopic detection of a neoplasm allows us to remove it and send to the pathology laboratory for definitive histological diagnosis. Moreover, snare polypectomy can be a radical treatment for dysplastic polyps without stromal axis and basal membrane infiltration. We therefore conclude that colonoscopy allows not only early diagnosis of colonic neoplasms, but also radical curative treatment in the early stages. PMID- 15553439 TI - [Iatrogenic injuries to the colon]. AB - The authors examine a number of aetiological, diagnostic and therapeutic aspects of iatrogenic injuries to the colon on the basis of an analysis of their own experience and a review of the literature. They highlight the low incidence of this kind of injury, which accounts for only 0.3% of all visceral injuries. Endoscopy and laparoscopic surgery are the most important causes of iatrogenic injuries to the colon. The authors conclude that these injuries are less serious and have a better prognosis than other kinds of colon perforations, because the colon is "clean" and it is possible to obtain an early diagnosis. Exceptions are lesions that occur in the course of a barium enema since these are associated with a high mortality rate which in some series may be as high as 100% of cases owing to the high toxicity of the barium contrast medium. PMID- 15553440 TI - [Preoperative radio-chemotherapy in the treatment of locally advanced rectal cancer]. AB - Rectal cancer is characterised by a substantial incidence of recurrences despite radical surgical treatment. The combination of preoperative radio- and chemotherapy has afforded functional and prognostic advantages through the prospect it offers of performing a greater number of conservative operations and the enhanced control of locoregional recurrences it allows. In our institute we treated 27 patients with locally advanced rectal cancer over the period from January 1997 to December 2002. All 27 patients underwent preoperative radiochemotherapy (45 Gy on the pelvis and 5-fluorouracil administered on the first and last 5 days of radiotherapy). The patients were then submitted to surgery consisting in 12 abdomino-perineal resections of the rectum, 14 anterior rectal resections and 1 Hartmann's resection. Tumour regression was complete in 22.22% of cases and minimal in 14.81%; 50% reduction was achieved in 22.22% and 50-80% reduction in 40-70% of cases. The toxicity was 14.91%. The incidence of local failure was 3.7% with a follow-up of 52 months. In this series, preoperative radio-chemotherapy proved to be a powerful means of downstaging the tumours and of controlling local failure. PMID- 15553441 TI - Meckel's diverticulum: a neglected (or deliberately ignored) entity. AB - Meckel's diverticulum is the most common of all the possible abnormalities that incomplete obliteration of the omphalo-mesenteric duct can produce. Often it is not sought during surgery performed for other abdominal diseases, e.g. cases of appendicitis. In these situations searching for and treating Meckel's diverticulum may be regarded as simply a waste of time or unjustified because its surgical treatment is not a completely safe procedure. In our opinion the systematic search for, and treatment of, Meckel's diverticulum during surgery performed for other abdominal conditions should always be performed. Elective resection of the diverticulum is characterised by fewer postoperative complications than emergency surgery. Laparoscopy would appear to be safe and effective in the treatment of this pathology, even in cases of severe diverticulitis. We report on our last five years' experience with the surgical treatment of this rare pathology, mainly occurring in previously appendectomised patients. One aim of this study was to demonstrate the possible benefits of the laparoscopic approach, focussing attention on the advisability of searching for and resecting Meckel's diverticulum when discovered incidentally during abdominal interventions performed for other pathologies. PMID- 15553442 TI - [Doppler-guided transanal haemorrhoidal dearterialisation]. AB - The usual surgical treatment for haemorrhoids consists in excision of the piles and ligation of the hemorrhoidal plexus, with considerable postoperative pain. A new, less invasive technique has been introduced, called transanal haemorrhoidal dearterialisation. This technique consists in Doppler-guided ligation of the distal branches of the superior rectal arteries (3 to 6) 2-3 cm above the pectinate line. Arterial ligation causes reduction of blood flow to, and decongestion of, the haemorrhoidal plexus. From January 2000 to September 2003, we performed transanal haemorrhoidal dearterialisation in 138 patients. Patients experienced no pain in the immediate postoperative period. The follow-up revealed good outcomes. The transanal haemorrhoidal dearterialisation procedure can be considered a safe, effective, painless and quick method of curing haemorrhoidal disease. Its indications are extensive. The success rate is approximately 90%, but may be lower for grade 4 haemorrhoids. PMID- 15553443 TI - [Transfixed stitches for the treatment of haemorrhoids]. AB - Nowadays the proctologist has the opportunity to perform various different surgical techniques for the treatment of hemorrhoids. Circumferential mucosectomy with a stapler, diathermic hemorrhoidectomy with high frequency devices, and the HLA doppler II system have significantly modified the classical indications for the treatment of the disease. There is, however, still no general consensus as to the indications for the use of each of these techniques in clinical practice, giving rise to confusion among specialists and an inappropriate use of health care resources. For these reasons the authors propose a new technique for the treatment of haemorrhoids based upon a new classification system, named PATE 2000 Sorrento. The transfixed correction of haemorrhoids makes it possible to treat third degree internal piles in association with the Milligan-Morgan procedure for fourth degree piles in a simple, definitive and safe session. This technique seems to offer a valid alternative to stapled hemorrhoidectomy, on the one hand, and classical open or closed hemorrhoidectomy combined with rubber band ligation, on the other. The authors describe the technical feasibility of their technique and underline the good clinical results obtained in their initial experience with 20 consecutive patients. PMID- 15553444 TI - Femoral hernia repair with Bard Mesh Dart Plug. AB - Several tension-free femoral hernia repair techniques are currently available with different approaches and prosthetic devices, all of which, however, largely yielding excellent results. The aim of this report was to describe the technical aspects of femoral hernia repair via an infrainguinal approach using the Bard Mesh Dart Plug, and to evaluate the short- and medium-term results in our experience. From May 2000 to December 2003, 25 patients (F/M ratio 2.1:1; mean age 66.4 years) underwent femoral hernia repair with the Bard Mesh Dart Plug. Fourteen patients (56%) were operated on as emergency cases for irreducible hernias and in 3 cases a bowel resection under general anaesthesia was necessary. The other 11 patients underwent surgery on an outpatient day surgery basis under local or spinal anaesthesia. Topical antibiotic prophylaxis was administered in all cases, while short-term prophylaxis with ceftazidime was limited to emergency patients. The main phases of the operation are described. The mean operative time was 40 minutes and the patients were mostly discharged within 2 hours of surgical treatment. No general or local intraoperative complications were registered. Postoperative mortality occurred in a single case (a 98-year-old woman, ASA IV, affected by strangulated hernia). Postoperative morbidity included 4 seromas and 1 haematoma. No recurrences were observed over a mean follow-up period of 24 months (range: 5-46 months). The femoral hernia repair with the Bard Mesh Dart Plug is a simple, safe, fast and effective procedure which can often be carried out in the day surgery setting. It can therefore be regarded as a valid alternative to other tension-free techniques. PMID- 15553445 TI - Giant bilateral ovarian cysts in an adolescent masked by obesity and mimicking ascites: a case report. AB - Ovarian cysts are a common pathology after the 4th decade of life. We can find either smaller functional, non-neoplastic ones (belonging to the follicular and luteinic varieties) or larger tumoral cysts, which, however, are usually benign. These may be of the serous or mucinous type and can sometimes reach really large sizes. Reports of giant ovarian manifestations were more frequent a few decades ago. Prior to the advent of modern radiological, ultrasonographic, tomographic and magnetic resonance imaging techniques, diagnosis was often difficult. Nevertheless, even today, in some cases (as a result of pronounced obesity, for example, associated perhaps with diagnostic negligence), cases of giant ovarian cysts may still be encountered. We report the case of a (previously obese) 19 year-old female, admitted to our hospital for presumed ascites, identified and ultrasonographically misdiagnosed by her gynaecologist. The patient was, in fact, suffering from giant serous cystoadenomas in both ovaries. PMID- 15553446 TI - [Aggressive angiomyxoma of the pelvis and perineum: a case report and review of the literature]. AB - Aggressive angiomyxoma is a rare mesenchymal tumour arising from soft tissue of the pelvis and perineum. Other features of this tumour are non-specific clinical signs, local invasion, a high local recurrence rate, and recurrence long after initial excision. We provide a case report with a review of the literature. A 57 year-old woman with a large bulky mass in the perineum was admitted to our institution. She had previously undergone iterative surgery for excision of pelvic and perineal masses. The preoperative diagnostic procedure is described as well as the therapeutic surgical approach (a double simultaneous laparotomic and perineal approach was opted for). The surgical procedure was difficult, time consuming and dangerous owing to an extensive area of hard retroperitoneal sclerosis involving the low urinary viscera, the vaginal stump and the mesorectum. Radical excision of the mass favoured the re-establishment of the normal pelvic anatomy, resolution of the preoperative symptoms and complete recovery of working capability. Surgical excision is the gold standard in the treatment of aggressive angiomyxoma. This benign neoplasm may sometimes present a malignant course owing to involvement of pelvic viscera. It is also associated with a high late recurrence rate due to local aggressiveness, and longterm follow up is therefore necessary. PMID- 15553447 TI - Spigelian hernia: a rare lateral ventral hernia. AB - The authors take a treated clinical case as a starting point to consider lateral ventral hernia, generally referred to as "Spigelian hernia". Such hernias are rare (1-2% of all hernias), with a slightly higher incidence in the female sex. Obesity and multiparous status are known to be predisposing factors. Instrumental tests of great importance in terms of specificity and definition such as ultrasonography and computed tomography are available, in cases of diagnostic doubt, for a pathology that in any case has to be detected early in order to avoid possible complications. The treatment is essentially surgical. A personal variant of the surgical repair technique is described with the use of prosthetic material, which guarantees a better result in terms of strength and resistance compared to simple repair surgery. PMID- 15553448 TI - [Congenital cystic bile duct dilatation: an incidental intraoperative finding]. AB - In adulthood, choledochal cysts are often surprisingly discovered during cholecystectomy. We report here on a case of an unsuspected congenital type-IA biliary cyst, according to Todani's classification encountered during laparoscopic cholecystectomy in a 30-year-old woman complaining of acute abdominal pain with an unremarkable preoperative workup. The well-known risk of developing cystic cancer, mainly in the adult, means that an excisional operation is indicated to prevent such complications. Cyst excision and Roux-en-Y hepaticojejunostomy is the definitive treatment of choice, performed at a later stage after radiological and endoscopic confirmation, and after obtaining the patient's consent. Cholangio-MR, and ERCP have proved to be extremely useful for adequate identification of this kind of lesion. PMID- 15553450 TI - [Historical evolution of the sentinel lymph node]. AB - The sentinel node is the first lymph node reached by metastasising cells from a primary tumour. Numerous studies have confirmed and demonstrated the reliability of the sentinel node biopsy hypothesis, on account of the high identification rate and overall accuracy of dye-guided and radio-guided procedures. It is remarkable that the concept of lymphatic mapping was not introduced until the end of the twentieth century. The development of the dynamic technique of intraoperative lymphatic mapping in the 1990s resulted in general acceptance of the sentinel node theory. The current literature suggests that sentinel node biopsy may eventually replace axillary dissection as the nodal staging procedure of choice in early breast cancer. This report describes the history and validation of the sentinel node biopsy technique, with particular reference to breast cancer. PMID- 15553449 TI - Endometriosis in an episiotomy scar: a case report. AB - Endometriosis is defined as the presence of functioning endometrial tissue in an anatomical location other than the uterine cavity. The episiotomy scar is a fairly rare site for endometriosis. The authors present the case of a 42-year-old woman referred 7 years ago for the development of a tender perianal mass at the episiotomy site associated with perianal pain and pruritus which varied in relation to menses, becoming most intense just before and immediately after the onset of menstrual bleeding. Anal endosonography performed with a B&K mechanical probe rotating through 360 degrees with a frequency of 7 and 10 Mhz showed a hypoechoic area with a diameter of 3 cm not involving the external sphincter and extending from the perianal skin to the mid third of the anal canal. Proctosigmoidoscopy findings were normal. A complete local excision was performed. Complete surgical excision of perineal endometriomas should be curative. Recurrence, supposedly due to incomplete removal, usually appears within one year. PMID- 15553451 TI - [Historical aspects of a frequent anal disease: haemorrhoids]. AB - In this report the authors outline the main historical aspects of haemorrhoids after a thorough review of the literature. The first texts concerning haemorrhoids are the ancient Egyptian papyruses. The twenty centuries of the history of haemorrhoids are marked by multiple pathogenetic theories, and of these the vascular and the sliding of the anal mucosa are still valid. From an analysis of the historical texts and the most recent publications, we deduce that the principles of therapy are substantially unchanged since the days of Hippocrates. In conclusion, we can say that, as in ancient times, the therapy of haemorrhoids is based on three fundamental approaches: surgical treatment, ambulatorial non-surgical treatment and medical treatment. PMID- 15553452 TI - Enhanced nitrogen removal in the combined activated sludge-biofilter system of the Southpest Wastewater Treatment Plant. AB - In 1999 the existing activated sludge unit of the Southpest Wastewater Treatment Plant was supplemented by a two-stage biofilter system aiming for nitrification and post-denitrification. In this arrangement excess biomass of the filters is wasted through the activated sludge unit, facilitating backseeding, and recirculation of the nitrate-rich effluent of the N-filter serves for decreasing the methanol demand of the DN-filter and for saving aeration energy at the same time. The paper reports on the development of an ASM1-based mathematical model that proved to be adequate for describing the interactions in the combined system and was used to compare the efficiency of different treatment options. Full-scale results verified that backseeding may considerably improve performance. However, nitrification ability of the activated sludge unit depends on the treatment temperature and, if unexpected, can be limited by insufficient oxygen supply. The upgrading possibilities outlined may serve as a new perspective for implementation of combined activated sludge-biofilter systems. PMID- 15553453 TI - Upgrading and expansion of the Kappala wastewater treatment plant. Operational experiences and results. AB - Kappala Association has the responsibility to receive and treat wastewater from 11 municipalities situated just north of Stockholm in Sweden. Running a tunnel system, 60 km long, and a treatment plant meets this responsibility. The plant is situated in Lidingo, northeast of Stockholm. The load 2002 corresponds to about 520,000 p.e. During the 1990s the plant was upgraded and expanded to meet an increasing population and more stringent discharge limits. Nitrogen removal was introduced. The expansion was done in two steps; the first step comprised a new plant beside the old one both situated in rock. This part, which included the filtration step, was taken into operation in 1998. Then the old plant was upgraded to the same technical standard as the new part. His Majesty the King of Sweden inaugurated the whole new and upgraded plant in April 2000. A total of 1.3 billion Swedish crowns were invested corresponding to about 140 million euro. The plant's performance is good. There is no trouble keeping the discharge limits, 10 mg/l for BOD7 and total nitrogen and 0.3 mg/l for phosphorus. However the bio-P process has not been altogether successful. We haven't been able to fulfill our goal to reduce the use of iron sulphate. We will during 2003 go on in our investigations concerning running the plant with a combination of bio-P and chemical precipitation. PMID- 15553454 TI - Nutrient removal process development and full scale implementation at the 4 million p.e. main treatment plant of Vienna, Austria. AB - The Main Treatment Plant of Vienna is in extension for 4 million p.e. and very stringent nutrient removal requirements. The existing high rate BOD removal activated sludge plant (in operation since 1980) is extended by a second stage activated sludge plant and a newly developed flow scheme for nitrogen removal optimisation adaptable to the temperature variations over the year. For this plant pilot investigations have been performed for the development of a specific mathematical model (ASMV) and a specific aeration control strategy. The civil work of the extension is already finished and the installation of the equipment has started. Operation should start in 2004. The whole project will cost about [see symbol in text]264 million of which about one half is for civil work. The effluent standards correspond to the requirements for sensitive areas in EU Directive for Municipal Waste Water. The raw primary and excess sludge are incinerated after thickening and dewatering. This paper tries to condense the already existing literature with the construction progress and the cost situation. PMID- 15553455 TI - The Polish perspective on adopting EU standards for nitrogen removal at large WWTPs--case studies. AB - The most challenging issue for existing large WWTPs (>100,000 PE) in Poland will be achievement of the new effluent standards for total nitrogen. Consequently, reliable and accurate information concerning the dimensioning of anoxic compartments is necessary. This study focused on validating to what extent the denitrification rates determined from batch tests were comparable with the rates calculated based on a mass balance over a full-scale activated sludge reactor. The experiments were conducted at two large WWTPs in northern Poland: "Wschod" in Gdansk and "Debogorze" in Gdynia. Two types of batch tests were used to determine the denitrification capability of activated sludge. Lower nitrate utilization rates observed during the full-scale experiments could potentially result from the local disturbances such as nitrate limitation ("Wschod" WWTP) or oxygen penetration to the anoxic zone ("Debogorze" WWTP). These factors should be taken into consideration during the design phase of the anoxic compartments. PMID- 15553456 TI - Improvement of nitrogen removal at WWTP Zurich Werdhoelzli after connection of WWTP Zurich-Glatt. AB - Optimisation of nitrifying activated sludge plants towards nutrient removal (denitrification and enhanced P-removal) leads to a substantial reduction of operating costs and improves effluent and operating conditions. At WWTP Zurich Werdhoelzli, initially designed for nitrification only, an anoxic zone of 28% of total activated sludge volume was installed and allowed 60% nitrogen elimination besides several other optimisations. In 2001 the operation of WWTP Zurich-Glatt was stopped and the wastewater was connected to WWTP Werdhoelzli. To improve nitrogen removal, WWTP Werdhoelzli co-financed two research projects; one for separate digester supernatant treatment with the anammox process operating two SBRs in series and the other applying NH4 sensors for aeration control in order to decrease energy consumption and raise effluent quality. The results of both projects and the consequences for WWTP Werdhoelzli are discussed in this paper. PMID- 15553457 TI - Optimisation of Hamburg's wastewater treatment plants--three years of experience with the new concept. AB - In the end of the year 1999 a new concept for Hamburg's wastewater treatment plants was commissioned to cope with the additional load of 250,000 PE which has been connected to the existing plant serving 1.85 million population equivalents. The new concept consists of an additional tank for the storage of sludge liquor to avoid peak nitrogen concentrations in the effluent of the plant. This solution has been developed within a study assessing a wide range of concepts with costs between initial [see symbol in text] 100 million and the final construction costs of [see symbol in text]1 million. Here the dynamic simulation showed to be a very efficient planning tool. Within the first three years of operation the chosen solution showed to be able to fulfil the requirements. In 2002 the concept has been improved by a few more measures, mainly the equipment of one of the storage tanks as a SAT tank for further cost effective nitrogen removal. For a further improvement of the treatment capacity additional measures are planned to be carried out within the next years. PMID- 15553458 TI - Combined quantity management and biological treatment of sludge liquor at Hamburg's wastewater treatment plants--first experience in operation with the Store and Treat process. AB - Store and Treat (SAT) is a new concept for the management of ammonium-rich process waste waters at wastewater treatment plants. It combines the advantages of quantity management and separate biological treatment, whereby both operations are carried out in the same tank. Now the first full-scale application of that method was realized in Hamburg. As first experience shows the process can help to increase nitrogen removal and to reduce energy consumption. PMID- 15553459 TI - Hygienization of municipal sludge in automatically operated chamber filter presses with thermal vacuum drying. AB - This paper presents the state of the art of thermal vacuum drying in chamber filter presses for unattended automatic operation. The achieved results are exemplified by the treatment of the two stage digested combined primary, chemical and biological sludge created by the VEAS concept for nutrient removal from municipal wastewater at VEAS. The water removal rate in each stage of the drying process is described, with comments on the low energy needs. The advantages of one-sided heating, the capacity and the drying potential are discussed. The hygienization potential of the process is demonstrated by the effect on thermostable coliform bacteria, Escherichia coli, salmonella, the spores of sulfite reducing anaerobic bacteria, f-specific bacteriophages, the seeds of the weed, wild oat, Avena fatua, and the parasite eggs of the potato cyst nematode, Globodera rostochiensis. A more complete paper with the VEAS-concept is found on the VEAS homepage (www.veas.nu). PMID- 15553460 TI - A control strategy for reducing aeration costs during low loading periods. AB - The efficiency of the aeration system in a full-scale activated sludge basin with 3 separately controlled aeration zones was improved for the low loading period in summer. The air flow rate to each aeration zone is currently regulated to hold a preset dissolved oxygen concentration (DO). Four different DO setpoint combinations were tested, each one for a one week period, using dynamic off-gas testing to measure the standardised oxygen transfer efficiency (alphaSOTE). As the DO setpoints were lowered, the total air flow rate to the basin decreased initially. A low DO in the first zones slowed biomass activity and pushed the load towards the end of the aeration basin. The relationship between alphaSOTE and the specific diffuser flow rate qD is different for each zone. In Zone 1 there was a strong decrease in alphaSOTE as qD increased, while Zones 2 and 3 were fairly independent of qD, Zone 2 at a higher level than Zone 3. Aeration costs were reduced by 15% for the most efficient combination. To achieve even more savings, a control strategy adjusting oxygen transfer rates over the aeration basin to the necessary oxygen transfer rates is suggested. It is based on changing the DO setpoints to reach the lowest total air flow rate while meeting the effluent requirements. PMID- 15553461 TI - Clogging and cleaning of fine-pore membrane diffusers. AB - Causes for the increase of the pressure loss of fine bubble diffusers made of elastomers were investigated. Methods are described for removing clogging material from membrane diffusers and/or attenuating or even avoiding the formation of clogging deposits. On the basis of some practical examples the procedure is discussed and obtained reductions of pressure losses are specified. PMID- 15553462 TI - Biological coating of EPDM-membranes of fine bubble diffusers. AB - Biological coatings on EPDM-membranes are a problem on many large wastewater treatment plants, as the oxygen supply of the micro-organisms is no longer guaranteed. Investigations prove that the pressure loss and the Shore A-hardness of the EPDM-membranes increase while on the other hand their softener content decreases accordingly. The detected coatings on the membrane surfaces and in the slits or holes of the membranes show extra-cellular organic substances (EPS), which, compared with fibrillar/filamented EPS usually found on surfaces in wastewater treatment plants, are viscous to a much greater extent. As, besides primary organic parts (carbon), the coatings on the membranes as well as in the slits or holes also consist of inorganic constituents (magnesium, silicon, and others), the authors assume that, the separating agent (and also inactive filler) talcum (magnesium silicate), used when producing the membranes, supports at least a first beginning of the coating. Superfine dust constituents and fibres, input via the compressed air, will build up inside the coating and consequently lead to a gradual clogging of the holes or slits. Besides chemical cleaning measures, the exchange of the EPDM-membranes against membranes of silicone would also be a possible measure to solve this problem. The market will decide, if, in the future, a cleaning or an exchange of the EPDM-membranes against membranes of silicone will be applied, but it has to be considered that the loss of softener is irreversible. PMID- 15553463 TI - Full-scale application of the BABE technology. AB - Bio-augmentation can be used to obtain nitrification in activated sludge processes that operate at sub-optimal solid retention times. A side-stream process, the so-called BABE process that incorporates N-removal and augmentation of nitrifiers has been developed. The principle is to implement a nitrification reactor in the sludge return line, the so-called BABE reactor. This reactor can be fed with an internal N-rich flow (e.g. effluent from the sludge treatment). Hence the nitrification capacity of an activated sludge process can be augmented by the addition of nitrifiers cultivated in the BABE reactor. A full-scale test of the BABE technology has been at the treatment plant Garmerwolde in Groningen, the Netherlands. The set-up allowed comparing between three different lines: with the BABE reactor, without rejectwater and with untreated rejectwater. Based on this, the two important tasks (N-removal and inoculation) performed by the BABE reactor could be quantified. The results of the practical work in Garmerwolde showed a higher nitrification rate in the water line where the BABE reactor was implemented and also lower effluent ammonia. The experiments on a practical scale have demonstrated univocally the effective and stable operation of the BABE technology. In addition, sludge samples in different streams as well as from the BABE reactor were analysed with FISH technique. The FISH results illustrated the augmentation effect of the BABE reactor on the stream with the BABE reactor. A mathematical model, based on ASM1 model and implemented in AQUASIM was developed and used for simulating the treatment plant of Garmerwolde. The simulation results indicated that better effect of the BABE technology is expected at lower ambient temperatures and smaller volume of the BABE reactor. The BABE reactor could also allow for providing more space for de-nitrification in the main water line when nitrification is efficient enough. PMID- 15553464 TI - Six years of operation of a fluidised bed reactor for denitrification. AB - At Himmerfjarden wastewater treatment plant, a fluidised bed reactor for nitrogen removal has been operated since 1997. Despite its small footprint, the system enables a far-reaching nitrogen removal. The current nitrate reduction in the reactor is 95%. The reduction of total nitrogen at the wastewater treatment plant is 80-90% at normal operation. The concentration of nitrate in the effluent is easily controlled by changing the dose of carbon. As part of a series of full scale experiments, the plant has, for the last two years, been operated without denitrification for a couple of months during spring/summer, in order to benefit a favourable N/P-ratio in the recipient and to counteract the growth of nitrogen fixing blue-green algae, When resuming the dosage of carbon, full denitrification was re-established in about two weeks. Important factors to take into consideration when operating the fluidised bed reactor are the abrasive characteristics of the carrier material (sand), the increased concentration of suspended solids in the effluent, and the importance of a suitable N/P-ratio in the influent, as lack of phosphorus might cause an uncontrolled microbiological growth. PMID- 15553465 TI - Denitrification rate and carbon source consumption in full-scale wastewater filtration. AB - In response to new demands for increased removal of nitrogen and phosphorus, the Henriksdal and Bromma treatment plants, with hydraulic loads of 283,000 and 148,000 m3/d, respectively, built filtration steps as a final process step in the plants. The denitrification rates in a full-scale and in a pilot plant filter are calculated to 13.1 and 21.3 g (NO3+NO2)-N/(m3 x h), respectively, in the total filter bed after 2.5-24.2 and 16.0-28.0 h of operational time, and 6.4 and 18.7 g (NO3+NO2)-N/(m3 x h), respectively, after 1.0 and 0.1-0.9 h of operational time. In composite samples, the denitrification rate in the total filter bed is 10-20 g (NO3+NO2)-N/(m3 x h) in the full-scale filter. The average values for k = deltaCODf/deltaC(T) are 1.6 and around 3 in the total filter bed in steady state and in the beginning of the experiments, respectively, both in the full-scale and in the pilot plant study. The carbon source costs for reducing the concentration of nitrate nitrogen in the Bromma plant from 12 to 8 mg/l in the effluent are 117,400 EUR and 147,400 EUR with methanol and ethanol, respectively, as a carbon source. PMID- 15553466 TI - Optimisation of a large WWTP thanks to mathematical modelling. AB - Better controlling and optimising the plant's processes has become a priority for WWTP (Wastewater Treatment Plant) managers. The main objective of this project is to develop a simplified mathematical tool able to reproduce and anticipate the behaviour of the Tougas WWTP (Nantes, France). This tool is aimed to be used directly by the managers of the site. The mathematical WWTP model was created using the software WEST. This paper describes the studied site and the modelling results obtained during the stage of the model calibration and validation. The good simulation results have allowed to show that despite a first very simple description of the WWTP, the model was able to correctly predict the nitrogen composition (ammonia and nitrate) of the effluent and the daily sludge extraction. Then, a second more detailed configuration of the WWTP was implemented. It has allowed to independently study the behaviour of each of four biological trains. Once this first stage will be completely achieved, the remainder of the study will focus on the operational use of a simplified simulator with the purpose of optimising the Tougas WWTP operation. PMID- 15553467 TI - Modeling of a large-scale wastewater treatment plant for efficient operation. AB - Environmental legislations in the Western world impose stringent effluent quality standards for ultimate protection of the environment. This is also observed in Turkey. The current paper presents efforts made to simulate an existing 0.77 million m3/day conventional activated sludge plant located at Ankara, AWTP. The ASM1 model was used for simulation in this study. The model contains numerous stoichiometric and kinetic parameters, some of which need to be determined on case by case bases. The easily degradable COD (S(S)) was determined by two methods, physical-chemical and respirometric methods, namely. The latter method was deemed unreliable and rejected in the further study. Dynamic simulation with SSSP program predicted effluent COD and MLSS values successfully while overestimating OUR. A complete fit could only be obtained by introducing a dimensionless correction factor (etaO2 = 0.58) to the oxygen term in ASM1. PMID- 15553468 TI - A guideline for simulation studies of wastewater treatment plants. AB - A guideline for simulation studies of wastewater treatment plants is proposed. The aim of the HSG-guideline is to define a reference quality level that helps to make the results of simulation studies comprehensible and comparable and therefore increases the quality and reliability of mathematical modelling in wastewater treatment. The paper gives a summary of the HSG-guideline, written by a group of university members from Germany, Austria and Switzerland. PMID- 15553469 TI - Performance intensification of Prague wastewater treatment plant. AB - Prague wastewater treatment plant was intensified during 1994--1997 by construction of new regeneration tank and four new secondary settling tanks. Nevertheless, more stringent effluent limits and operational problems gave rise to necessity for further intensification and optimisation of plant performance. This paper describes principal operational problems of the plant and shows solutions and achieved results that have lead to plant performance stabilisation. The following items are discussed: low nitrification capacity, nitrification bioaugmentation, activated sludge bulking, insufficient sludge disposal capacity, chemical precipitation of raw wastewater, simultaneous precipitation, sludge chlorination, installation of denitrification zones, sludge rising in secondary settling tanks due to denitrification, dosage of cationic polymeric organic flocculant to secondary settling tanks, thermophilic operation of digestors, surplus activated sludge pre-thickening, mathematical modelling. PMID- 15553470 TI - Sewage-treatment under substantial load variations in winter tourism areas--a full case study. AB - The sewage-load variations in winter tourism areas are characterized by sudden increases--in the range of a factor two to three--within only a few days at the start and the end of the tourist season, especially at Christmas. The sudden load increases occur during periods of low wastewater temperatures, which is an additional demanding factor with respect to nitrogen removal. A full case study was carried out at WWTP Saalfelden, which is located near one of Austria's largest skiing resorts. The plant is designed for 80,000 PE and built according to the HYBRID-concept, which is a special two stage activated sludge process for extensive nutrient removal. PMID- 15553471 TI - Reducing the total discharge from a large WWTP by separate treatment of primary effluent overflow. AB - At many large wastewater treatment plants (WWTPs) the increased hydraulic load, caused by combined sewer systems during storm events, results in primary effluent overflow when the capacity of further treatment is exceeded. Due to stringent effluent standards, regulating the total discharge from the WWTPs, the Rya WWTP in Goteborg and the Sjolunda WWTP in Malmo will have to reduce the impact of primary effluent overflow. Separate, high rate, precipitation processes operated only during high flow conditions have been investigated in pilot units at the two WWTPs. Precipitation in existing primary settlers operated at a surface loading of 3.75 m/h removed phosphorus to 0.35 mg/l. The Actiflo process was also shown to remove suspended solids and phosphorus well. BOD was reduced by 50-60%. With such processes the overall effluent concentrations from the plants can be reduced significantly. Key upgrading features are small footprints, short start up time and high efficiency. PMID- 15553473 TI - Biological solution to storm water? AB - Standard practice in Flanders is to limit the hydraulic capacity of sewage treatment works to 6Q14 (Q14= 1.7 dry weather flow Q(DWF)). A maximum of 3Q14 is treated biologically, while the excess flow undergoes only physical treatment in storm tanks. This practice has been challenged by a new high-flow activated sludge operation concept, consisting of the treatment of the full storm sewage flow in the biological train and of the use of the storm tanks as additional secondary clarifiers. After successful testing in two installations, 56 works of different sizes and types were switched to high-flow activated sludge operation from 1999 to 2002. This paper reports on progress and experiences gained since then. The analysis focuses on the parameters subject to regulatory discharge (BOD, COD, suspended solids, total nitrogen and total phosphorus) plus ammonia. Special attention is paid to the performance of the clarification and of the nitrification processes. The results indicate that high-flow biological treatment provides a substantial reduction in wet weather discharges while maintaining acceptable process operating conditions. PMID- 15553472 TI - Selection of an ultraviolet disinfection system for a municipal wastewater treatment plant. AB - As part of an expansion to an average flow of 45.9 million gallons per day (174 mld), the Ypsilanti Community Utilities Authority wastewater treatment plant in the State of Michigan, USA, elected to install ultraviolet disinfection as a replacement for the existing chlorination process. This paper presents a unique methodology used in selecting the best system based on not only the life cycle costs, and O & M considerations but also the participation of the stakeholders. The Team members consisted of representatives of all departments at the Authority, and these Team members made the decision. The Team evaluated all criteria in the office, which was followed by verification at selected sites with similar types of equipment. The selected equipment then was pilot tested for validation of the dose-kill relationship under normal operation and also under reduced irradiation conditions. A low-pressure, high intensity system was selected, based on life-cycle cost, reliability, safety, and ease of operation. This paper describes the unique methodologies used in making that decision. The full-scale system is scheduled for start-up in Spring 2003. PMID- 15553474 TI - Influence of sludge properties and hydraulic loading on the performance of secondary settling tanks--full-scale operational results. AB - Full-scale investigations at a WWTP with a two-stage secondary settling tank process revealed relationships between significant operating parameters and performance in terms of effluent suspended solids concentration. Besides common parameters (e.g. surface overflow rate and sludge volume loading rate) feed SS concentration and flocculation time must be considered. Concentration of the return activated sludge may help to estimate the performance of existing secondary settling tanks. PMID- 15553475 TI - Dynamics in maximal settling capacity in an activated sludge treatment plant with highly loaded secondary settlers. AB - Secondary settling dynamics at maximal capacity were investigated at a full scale wastewater treatment plant which utilizes a unique process solution incorporating pre-denitrification with post-nitrification in nitrifying trickling filters. Since nitrogen removal is greater when more secondary effluent is recirculated to the trickling filters, the secondary settlers generally operate at close to their maximal capacity. The settling and flocculation properties of the activated sludge are therefore a major capacity-determining factor for plant operation. Due to the short sludge age, the flocculation properties, with respect to both thickening and clarification, can change quickly. The dynamics in these changes were studied and the factors that determine the maximal settling capacity were assessed. Solids flux curves were constructed from batch settling tests and compared with the actual maximal settling capacities. PMID- 15553476 TI - Comparison of the 1D flux theory with a 2D hydrodynamic secondary settling tank model. AB - The applicability of the 1D idealized flux theory (1DFT) for design of secondary settling tanks (SSTs) is evaluated by comparing its predicted maximum surface overflow (SOR) and solids loading (SLR) rates with that calculated from the 2D hydrodynamic model SettlerCAD using as a basis 35 full scale SST stress tests conducted on different SSTs with diameters from 30 to 45m and 2.25 to 4.1 m side water depth, with and without Stamford baffles. From the simulations, a relatively consistent pattern appeared, i.e. that the 1DFT can be used for design but its predicted maximum SLR needs to be reduced by an appropriate flux rating, the magnitude of which depends mainly on SST depth and hydraulic loading rate (HLR). Simulations of the sloping bottom shallow (1.5-2.5 m SWD) Dutch SSTs tested by STOWa and the Watts et al. SST, all with doubled SWDs, and the Darvill new (4.1 m) and old (2.5 m) SSTs with interchanged depths, were run to confirm the sensitivity of the flux rating to depth and HLR. Simulations with and without a Stamford baffle were also done. While the design of the internal features of the SST, such as baffling, have a marked influence on the effluent SS concentration for underloaded SSTs, these features appeared to have only a small influence on the flux rating, i.e. capacity, of the SST, In the meantime until more information is obtained, it would appear that from the simulations so far that the flux rating of 0.80 of the 1DFT maximum SLR recommended by Ekama and Marais remains a reasonable value to apply in the design of full scale SSTs--for deep SSTs (4 m SWD) the flux rating could be increased to 0.85 and for shallow SSTs (2.5 m SWD) decreased to 0.75. It is recommended that (i) while the apparent interrelationship between SST flux rating and depth suggests some optimization of the volume of the SST, that this be avoided and that (ii) the depth of the SST be designed independently of the surface area as is usually the practice and once selected, the appropriate flux rating is applied to the 1DFT estimate of the surface area. PMID- 15553477 TI - Lamella separators in the upgrading of a large urban sewage treatment plant. AB - Wupperverband is using lamella separators for the upgrading of its Kohlfurth sewage treatment plant that is currently in progress. The lamellae positioned at the outlet of the biological treatment stage already remove part of the biomass in the activation basin and prevent it from reaching the final clarification stage. This preliminary separation system reduces solids concentration in the biological treatment system without negative impact on final clarification and therefore also lowers the basin capacity needed, with positive effects on costs. This article gives an overview of the separation performance achieved. PMID- 15553478 TI - Commissioning and operational experiences for the 160ML/d Woodman Point Sequencing Batch Reactor--control of settleability and denitrification using bioselectors. AB - Achieving and maintaining good biomass settling characteristics is a critical process design objective for any activated sludge wastewater treatment plant (WWTP), whether intermittent or continuous technology. One way of ensuring good sludge settleability in intermittent WWTPs is the incorporation of bioselectors in the process. A bioselector is essentially a small discrete reactor volume designed primarily for carbon absorption, in which activated sludge organisms are exposed to a high substrate concentration for a relatively short time. It is normally very much smaller than an anoxic zone and the activated sludge recycle is only a fraction of that typically adopted in continuous plants. With proper conditioning, recycled biomass rapidly absorbs and stores soluble organic wastewater components before transfer to the main treatment basin. This absorption and storage mechanism, and careful management of aeration throughout the intermittent treatment cycle, plays a crucial role in many subsequent growth and treatment processes, including sludge floc formation, denitrification and biological phosphorus removal. This paper examines some design considerations, and reviews the benefits of bioselectors by reference to the commissioning and initial operation of the new 160ML/d Woodman Point Sequencing Batch Reactor in Perth, Western Australia. The applicability of bioselectors in continuous plants is discussed. PMID- 15553479 TI - North American performance experience with anoxic and anaerobic selectors for activated sludge bulking control. AB - Obtaining reliably high compaction characteristics and low SVI values has enormous economic consequences on activated sludge plant design and operation. Now that use of selectors has become more common for achieving low SVI values, the assessment of the full-scale performance characteristics of activated sludge plants incorporating selectors has high value for the design and operation of future plants. Treatment plant effluent quality is seldom analyzed solely on an average basis and the impact peak events have on effluent quality must be considered in design and operation. Descriptive statistics should also be used to assess selector performance, so that the influence of infrequently occurring values on plant design and operation can be assessed. The performance of 21 activated sludge plants incorporating selectors was evaluated. All the selector plants for which post installation data were available showed that the operating SVI values were significantly improved. One plant studied in detail showed that final effluent quality was significantly better after the selector installation. As a group, activated sludge plants with anaerobic selectors outperform those with anoxic selectors. Dissolved oxygen control is just as important in activated sludge plants with selectors as in conventional activated sludge plants. PMID- 15553480 TI - A new side stream process for easily degradable industrial waste waters to avoid sludge bulking. AB - A new treatment scheme for the treatment of easily biodegradable industrial waste waters has been developed. The side stream treatment of dairy waste water with the excess sludge from the domestic treatment line of the regional treatment plant Bad Voslau has been operated successfully for a period of three years during which the industrial load stemming from the dairy increased from 800 kg COD/d to 2,500 kg COD/d with peak loads up to 5,000 kg/d. Despite of the increased load to the treatment plant the total aeration tank volume had not been increased. This treatment is performed in an existing aeration tank of the WWTP (V = 1,800 m3) which is now used as contact tank for the combined aeration of dairy waste water and excess sludge from the domestic treatment line (volume aeration tank = 15,000 m3). In this tank the easily degradable substrate from the industrial waste is mainly adsorbed to the biological sludge and after a mechanical dewatering transferred to the anaerobic digester where it yields in an increased gas production. The filtrate of the dewatering process is completely free from biodegradable material and can without danger of bulking be fed to the aeration tank of the domestic treatment line. The new process has proven to be extremely flexible since already now daily peak loads exceeding the design load by more then 60% could be treated in the plant without any problems. Compared to other alternatives for the dairy waste water treatment that were investigated during this study, the new side stream process is very advantageous. No other pre treatment process for industrial waste water could have been operated under comparable loading conditions without severe operating problems. PMID- 15553481 TI - Cost-effective solutions for sewage treatment in developing countries--the case of Brazil. AB - Cost-effective solutions are a must in developing countries, not only regarding investment costs, but also in respect to technology and operating practices. With these two goals in mind, in Brazil a particular effort has been directed for the development and application of the Chemical Enhanced Primary Treatment (CEPT) process and of the Upflow Anaerobic Sludge Blanket (UASB) process, both followed by complementary secondary treatment. Both technologies are under current expansion in Brazil. Large CEPT plants have been designed and built, up to 3.7 m3/s average design flow, as well as large UASB reactors, up to 3.0 m3/s average design flow. The applied technologies are cost-effective: they present low investment and efficiencies of BOD removal of up to 50% to 70%. They allow the plant construction in steps, an initial phase with efficiency over the usual primary treatment, and in order to achieve best effluent quality and meet legal water quality standards, a logic upgrade post-treatment can later on be implemented. The higher initial reduction of BOD and TSS also permits savings in construction and operational costs of secondary treatment, due to lower organic load and lower energy consumption. Sludge represents a particular point of attention: in the cases when the CEPT was used, Chemical Stabilisation of the Sludge (CSS) has also been practiced, eliminating the high construction costs of the digesters, all the plant staying chemically operated. In the cases when the UASB is used preceding secondary treatment, sludge can easily return to the anaerobic vessel, the costly sludge digestion unit being avoided. UASB reactors have practically no equipment in the anaerobic vessel, no energy consumption, low sludge production, and when applied in hot climates as in Brazil, heating devices are not required. The Brazilian experience, some particular cases, special comments on design and different secondary treatment processes are presented in this paper, as a contribution to the discussion of cost and benefits, a prime point to be considered. PMID- 15553482 TI - Economic and environmental optimization of phosphorus removal. AB - Do operating costs conform to environmental impact after introduction of charges on discharge of wastewater? A study on optimization of phosphorus removal at two Danish wastewater removal plants shows that this is actually the case. By measurement of inlet and outlet concentrations and of chemicals added it was possible to determine the relationships between chemical dosing and phosphorus discharge and thus calculate the operational cost and environmental impact of different dosing/discharge levels. PMID- 15553483 TI - Performances of three R-AN-D-N wastewater treatment plants in the Czech Republic. AB - The evolution of Czech standards requires higher efficiency of nutrient removal from municipal wastewaters. At the beginning of the last decade of 20th century, a new activated sludge configuration called R-AN-D-N process has been described, successfully tested and now largely used at several wastewater treatment plants (WWTP) in the Czech republic. The main feature of the R-AN-D-N process is the introduction of a regeneration zone in sludge recycle, which enables to increase sludge age in the system without any substantial increase in WWTP volume. Performances of three Czech large WWTP with R-AN-D-N configuration have been monitored and compared within a period of one and a half years. The results confirmed excellent nutrient removal efficiency for wastewaters with different proportion between sewage and industrial effluents. Two of three monitored WWTP received wastewaters from breweries (Budweiser and Pilsner Urquell). The settleability of activated sludge from all three WWTP was correct, with SVI values usually ranging from 50 to 150 ml/g. Monitoring of sludge composition indicated proliferation of several filamentous bacteria, particularly types 0581, 0092 and M. parvicella. No severe bulking events were observed. Finally, the operational costs expressed in CZK (Czech crown: 1 CZK = [see symbol in text]0.0322) per cubic metre of treated sewage or per capita amounts respectively from 2.24 to 6.52, and from 285 to 342. PMID- 15553484 TI - Realisation of the EU Directive 91/271/EWG in Germany technical and economic effects from the perspective of an operator of large wastewater treatment plants. AB - In the Federal Republic of Germany, the requirements on the effluent quality which wastewater treatment plants (WWTPs) with a design value larger than 100,000 PE must achieve in regard to the parameter Nanorg were increased by a national regulation on 01/08/2002. The reason for this action was a pending threat of the European Court because of the insufficient realisation of the EU Municipal Wastewater Directive (91/271/EWG). The report at hand describes the consequences of such an increase of the requirements for operators of WWTPs, using the example of Emschergenossenschaft and Lippeverband. The currently applicable demands in Germany are compared to those valid on a European level, and the current construction and performance state of the WWTPs operated by Emschergenossenschaft and Lippeverband is presented. The secure compliance with the increased requirements necessitates optimisation measures, but also construction measures; these are described in regard to their technical realisation and the prospective costs. PMID- 15553485 TI - Benchmarking of municipal waste water treatment plants (an Austrian project). AB - An Austrian research project focused on the development of process indicators for treatment plants with different process and operation modes. The whole treatment scheme was subdivided into four processes, i.e. mechanical pretreatment (Process 1), mechanical-biological waste water treatment (Process 2), sludge thickening and stabilisation (Process 3) and further sludge treatment and disposal (Process 4). In order to get comparable process indicators it was necessary to subdivide the sample of 76 individual treatment plants all over Austria into five groups according to their mean organic load (COD) in the influent. The specific total yearly costs, the yearly operating costs and the yearly capital costs of the four processes have been related to the yearly average of the measured organic load expressed in COD (110 g COD/pe/d). The specific investment costs for the whole treatment plant and for Process 2 have been related to a calculated standard design capacity of the mechanical-biological part of the treatment plant expressed in COD. The capital costs of processes 1, 3 and 4 have been related to the design capacity of the treatment plant. For each group (related to the size of the plant) a benchmark band has been defined for the total yearly costs, the total yearly operational costs and the total yearly capital costs. For the operational costs of the Processes 1 to 4 one benchmark ([see symbol in text] per pe/year) has been defined for each group. In addition a theoretical cost reduction potential has been calculated. The cost efficiency in regard to water protection and some special sub-processes such as aeration and sludge dewatering has been analysed. PMID- 15553486 TI - The effect of public or private structures in wastewater treatment on the conditions for the design, construction and operation of wastewater treatment plants. AB - Organised in public or private structures, wastewater services have to cope with different framework conditions as regards planning, construction, financing and operation. This leads quite often to different modes of management. In recent years there has been a push for privatisation on the water sector in general, the reasons for which are manifold, ranging from access to external know-how and capital to synergistic effects through integration of wastewater treatment into other tasks of similar or equal nature. Discussed are various models of public/private partnership (PPP) in wastewater treatment, encompassing for example the delegation of partial tasks or even the proportional or entire transfer of ownership of treatment facilities to private third parties. Decisive for high performance and efficiency is not the legal or organisational form, but rather the clear and unmistakable definition of tasks which are to be assigned to the different parties, customers and all other partners involved, as well as of clear-cut interfaces. On account of the (of course legitimate) profit-oriented perspective of the private sector, some decision-making processes in relation to project implementation (design and construction) and to operational aspects will differ from those typically found on the public sector. This does apply to decisions on investments, financing and on technical solutions too. On the other hand, core competencies in wastewater treatment should not be outsourced, but remain the public bodies' responsibility, even with 'far-reaching' privatisation models. Such core competencies are all efforts geared to sustainable wastewater treatment as life-supporting provision for the future or as contribution to the protection of health and the environment and to the development of infrastructure. Major areas of wastewater treatment and other related tasks are reviewed. The paper concludes with a list of questions on the issue of outsourcing.